PMID- 15332110 TI - Exudative bullous retinal detachment after peripheral blood stem cell transplantation. PMID- 15332111 TI - Does vitreoretinal surgery under local anaesthesia result in missed fellow eye pathology? AB - PURPOSE: There has been a significant increase in the number of vitreoretinal procedures being performed under local anaesthesia over the past few years. This trend is expected to continue. This study was performed to investigate whether by undertaking retinal detachment surgery under local anaesthesia fellow eye examination was compromised. DESIGN: This was a prospective, consecutive, blind, observational study. METHODS: SETTING: This study was performed at a tertiary referral vitreoretinal unit in a teaching hospital. STUDY POPULATION: In all, 108 consecutive patients undergoing retinal detachment surgery under general anaesthesia were included. OBSERVATION PROCEDURES: Patients were examined independently by different retinal surgeons pre- and intraoperatively. MAIN OUTCOME MEASURES: The findings of the two examiners were compared and differences were analysed. RESULTS: There were 108 patients in this study, 57 of these were males and 51 females. The mean age was 59.01 years (range 16-91). Of these 108 eyes, 48/108 (49.08%) the preoperative examination was regarded as unsatisfactory by the examiner. Over 34% of eyes had fellow eye pathology when examined preoperatively but there were nine (8.33%) eyes in which additional lesions were found intraoperatively. CONCLUSION: General anaesthesia should be considered for patients in whom preoperative fellow eye examination is unsatisfactory. PMID- 15332112 TI - A neural-specific splicing event generates an active form of the Wiskott-Aldrich syndrome protein. AB - Actin polymerization is required for cellular events such as podosome, lamellipode or filopode formation in migrating cells, and members of the Wiskott Aldrich syndrome protein (WASP) family have essential roles in regulating actin dynamics at the cell leading edge. However, WASP proteins need first to be activated in order to be able to target actin polymerization. Here, we show the occurrence of a neural-specific splicing event, which is favoured by the nuclear orphan receptor chicken ovalbumin upstream promoter-transcription factor I, and generates a truncated WASP protein deleted of exon 2-encoded amino acids. This deletion relocates the protein to the plasma membrane and induces the formation of actin-rich podosome-like structures that also contain paxillin and vinculin. Furthermore, expression of the truncated protein in PC12 cells, as well as in primary neurons, stimulates neuritogenesis. These data underscore the importance of the neural-specific splicing of WASP RNA during development. PMID- 15332113 TI - Position of the CrPV IRES on the 40S subunit and factor dependence of IRES/80S ribosome assembly. AB - The cricket paralysis virus intergenic region internal ribosomal entry site (CrPV IGR IRES) can assemble translation initiation complexes by binding to 40S subunits without Met-tRNA(Met)(i) and initiation factors (eIFs) and then by joining directly with 60S subunits, yielding elongation-competent 80S ribosomes. Here, we report that eIF1, eIF1A and eIF3 do not significantly influence IRES/40S subunit binding but strongly inhibit subunit joining and the first elongation cycle. The IRES can avoid their inhibitory effect by its ability to bind directly to 80S ribosomes. The IRES's ability to bind to 40S subunits simultaneously with eIF1 allowed us to use directed hydroxyl radical cleavage to map its position relative to the known position of eIF1. A connecting loop in the IRES's pseudoknot (PK) III domain, part of PK II and the entire domain containing PK I are solvent-exposed and occupy the E site and regions of the P site that are usually occupied by Met-tRNA(Met)(i). PMID- 15332114 TI - The small chemical vacuolin-1 inhibits Ca(2+)-dependent lysosomal exocytosis but not cell resealing. AB - Resealing after wounding, the process of repair following plasma membrane damage, requires exocytosis. Vacuolins are molecules that induce rapid formation of large, swollen structures derived from endosomes and lysosomes by homotypic fusion combined with uncontrolled fusion of the inner and limiting membranes of these organelles. Vacuolin-1, the most potent compound, blocks the Ca(2+) dependent exocytosis of lysosomes induced by ionomycin or plasma membrane wounding, without affecting the process of resealing. In contrast, other cell structures and membrane trafficking functions including exocytosis of enlargeosomes are unaffected. Because cells heal normally in the presence of vacuolin-1, we suggest that lysosomes are dispensable for resealing. PMID- 15332115 TI - Robust infectivity and replication of Delta-24 adenovirus induce cell death in human medulloblastoma. AB - The diverse advanced treatment modalities currently available to children with medulloblastoma, including surgery and radiotherapy, are associated with deleterious side effects and often with an unfavorable prognosis. A mutant adenovirus, Delta-24, which has a 24-base pair deletion in the Rb-binding region of the E1A gene, demonstrates selective replication and oncolysis in various malignant phenotypes. Here we report the ability of Delta-24 to kill medulloblastoma cells. Flow cytometric analyses of cell receptors demonstrated expression of the coxsackie adenovirus receptor and RGD-related integrins in the assessed medulloblastoma cell lines. Infectivity assays using a replication deficient adenovirus to transduce the green fluorescence protein gene showed that the Delta-24 adenovirus infects 99% of Daoy and 46% of D283 Med medulloblastoma cells at a multiplicity of infection (MOI) of 50. Within 4 days after infecting medulloblastoma cells with Delta-24, a noticeable cytopathic effect was produced. Delta-24 induced a total cytopathic effect in Daoy and D283 Med medulloblastoma cells after 6 and 8 days of infection, respectively. In the infected population of cells, cell death correlated with the accumulation of cells in the S phase. At 5 days post-infection with 2.5 MOIs of Delta-24 adenovirus, the percentage of Daoy medulloblastoma cells in the S phase increased to 71.9+/-5.5%, compared with control values of 20.5+/-1.4%. The release of viral progeny was quantified as being increased by two orders of magnitude, indicating efficient replication of Delta-24 in medulloblastoma cells. This is the first report of the ability of oncolytic adenoviruses to infect and kill medulloblastoma cells, the findings of which suggest the potential efficacy of Delta-24 as a therapy for human medulloblastoma tumors. PMID- 15332116 TI - Inhibition of the NF-kappaB pathway enhances TRAIL-mediated apoptosis in neuroblastoma cells. AB - Neuroblastoma is the most common solid extracranial neoplasm in children and causes many deaths. Despite treatment advances, prognosis for neuroblastoma remains poor, and a critical need exists for the development of new treatment regimens. TNF-related apoptosis-inducing-ligand (TRAIL) induces cell death in a variety of tumors, but not in normal tissues. Moreover, TRAIL is nontoxic, making it a strong antitumor therapeutic candidate. We demonstrate that introduction of the TRAIL gene into neuroblastoma cell lines using an adenoviral vector leads to apoptotic cell death. RT-PCR and flow-cytometric analyses demonstrated that TRAIL's effect is mediated primarily via the TRAIL R2 receptor. As TRAIL can activate the nuclear factor-kappaB (NF-kappaB) signaling pathway, which can exert an antiapoptotic effect, we hypothesized that inhibition of NF-kappaB signaling may augment TRAIL's killing effects. TRAIL-mediated cell death was enhanced when neuroblastoma cells were simultaneously infected with a dominant-negative mutant of IkappaB kinase, a kinase essential for NF-kappaB activation. The combination of blockade of NF-kappaB signaling and expression of TRAIL induced apoptotic death in a greater proportion of SKNSH cells than did either treatment alone. Thus, concurrent inhibition of the NF-kappaB pathway and the induction of TRAIL mediated apoptosis may become a useful approach for the treatment of neuroblastoma. PMID- 15332117 TI - EDAG regulates the proliferation and differentiation of hematopoietic cells and resists cell apoptosis through the activation of nuclear factor-kappa B. AB - Erythroid differentiation-associated gene (EDAG) is considered to be a human hematopoiesis-specific gene. Here, we reported that downregulation of EDAG protein in K562 cells resulted in inhibition of growth and colony formation, and enhancement of sensitivity to erythroid differentiation induced by hemin. Overexpression of EDAG in HL-60 cells significantly blocked the expression of the monocyte/macrophage differentiation marker CD11b after pentahydroxytiglia myristate acetate induction. Moreover, overexpression of EDAG in pro-B Ba/F3 cells prolonged survival and increased the expression of c-Myc, Bcl-2 and Bcl-xL in the absence of interleukin-3 (IL-3). Furthermore, we showed that EDAG enhanced the transcriptional activity of nuclear factor-kappa B (NF-kappa B), and high DNA binding activity of NF-kappa B was sustained in Ba/F3 EDAG cells after IL-3 was withdrawn. Inhibition of NF-kappa B activity resulted in promoting Ba/F3 EDAG cells death. These results suggest that EDAG regulates the proliferation and differentiation of hematopoietic cells and resists cell apoptosis through the activation of NF-kappa B. PMID- 15332118 TI - Protective effects of nicotine against glutamate-induced neurotoxicity in PC12 cells. AB - This study aimed to assess whether nicotine prevented glutamate neurotoxicity in PC12 cells, and to identify the molecular mechanisms of any effects. The results showed that glutamate neurotoxicity in PC12 cells could be prevented by treatment with nicotine at concentrations of 10 nmol x l(-1) - 1 mmol x l(-1). This effect was in turn found to be inhibited by the application of the nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine. Nicotine significantly decreased the basal level of intracellular free Ca(+2) and enhanced the buffering action on Ca(+2) overload induced by high concentrations of glutamate (5 mmol x l(-1)). In addition, nicotine treatment up-regulated the mRNA and protein expression of apoptosis-related factors including bcl-2 mRNA and protein, but down-regulated the expression of bax mRNA and protein. It is concluded that the protective effects of nicotine against the neurotoxicity induced by glutamate are mediated by nAChRs, due to the increased buffering action on Ca(+2)and the modulation of apoptotic processes. PMID- 15332119 TI - DNA replication reaction in Xenopus cell-free system is suppressed by high pressure. AB - Previously, we demonstrated that when mouse erythroleukemia cells are exposed to a pressure of 80 MPa, the cell-cycle progression of S-phase cells is retarded. To examine the effects of high pressure on DNA replication, we used a Xenopus cell free system. From cell-cycle progression of sperm nuclei, it was found that sperm nuclei are stable to a pressure of 80 MPa, whereas egg extracts are susceptible to high pressure. Similarly, biotin-16-dUTP was incorporated into 80 MPa-treated sperm nuclei in pressure-untreated extracts, but not into naive sperm nuclei in 80 MPa-treated extracts. These results indicate that DNA replication in Xenopus cell-free system is suppressed by the susceptibility of the extracts to a pressure of 80 MPa. PMID- 15332120 TI - Site-specific excision or protection of an alpha A globin gene genomic site in apoptotic transformed chicken erythroblasts. AB - There is still no clarity on whether the endonuclease incisions in apoptotic cells are induced randomly in the genome or induced in some preferable sites. In order to evaluate the intensity of DNA fragmentation in the chicken alpha-globin domain, AEV-virus transformed chicken erythroblasts (HD3) were incubated in a serum free medium, and their DNA was Southern blotted and hybridised with probes representing different fragments of the domain. Probes corresponding to the upstream areas of the domain mostly hybridised with high molecular weight DNA. Unlike these, the probe corresponding to the 2 Kb BamHI-BamHI fragment, containing the alphaA globin gene (B18), revealed a 5 Kb band on the hybridisation autoradiographs. The probe to the neighbouring upstream fragment did not reveal this band, but it was clearly seen on hybridisations with a downstream 1 Kb BamHI-BamHI fragment. The intensity of the band increased with overall apoptotic DNA degradation, hence its appearance should be coupled to apoptosis. Hybridisation of BamHI-digested DNA with B18 probe revealed a shortening of the 2 Kb band in preparations of DNA from apoptotic cells. The presumable positions of the cuts correspond to the formerly described DNase hypersensitive sites in the domain. Slot-blot and Northern hybridisation of RNA extracted from apoptotic HD3 cells revealed that the excision of the area of the B18 gene is coupled to a decrease in the intensity of alphaA globin gene transcription. Transcription of the non-erythroid NIK gene, transcribed in the upstream part of the domain, did not depend on the level of apoptotic DNA fragmentation. PMID- 15332121 TI - Shoot regeneration from GUS-transformed tomato (Lycopersicon esculentum) hairy root. AB - To study the influence of genetic background on the transformation and regeneration of cultivated tomato plants, hairy root lines of tomato (Lycopersicon esculentum) were obtained by inoculating the hypocotyl explants of three tomato cultivars with the Agrobacterium rhizogenes strain DCAR-2, which harbors the pBI-121 binary vector. The Ri-T-DNA transformation into the plant DNA was confirmed by both of mikimopine and GUS assay analyses. The regeneration efficiency from hairy root explants was assessed. The data indicated that white embryonic calli were formed within two weeks in the presence of 2 mgl(-1) 2, 4-D plus 0.25 mgl(-1) kinetin. Adventitious shoots emerged from the embryonic callus in the presence of 1 mgl(-1) GA3 along with 0.5 mgl(-1) NAA. The regeneration frequency was higher in the cultivar UC-97, followed by Momotaro and then Edkawi. Molecular confirmation of the integration of the GUS gene into the hairy root derived plants genomes was done via PCR using GUS-specific primers and also using Southern blotting analysis. Our data shows that regeneration is possible from hairy roots of the cultivated tomato and this system could be used to produce transgenic tomato plants expressing the genes present in Agrobacterium rhizogenes binary vectors. PMID- 15332122 TI - The effect of antisense oligonucleotide treatment of plasma membrane Ca(+2) ATPase in PC12 cells. AB - Plasma membrane Ca(+2)-ATPase (PMCA), encoded by four separate genes, constitutes a high affinity system extruding Ca(+2) outside the cell. The nerve growth factor treated PC12 cell line possesses all four main PMCA isoforms. To evaluate the potential role of PMCA isoforms in the differentiation process, we transiently suppressed the expression of PMCA2 and 3 using the antisense oligonucleotides. In the transfected PC12 cells, we observed morphological changes, slowed neurite extension and diminished survival of the cells. The apparent transport activity and affinity of the calcium pump to Ca(+2) were lower in the cells with suppressed PMCA2 and 3 isoforms than in the control cells. Moreover, in the transfected PC12 plasma membranes, the calcium pump was insensitive to stimulation by calmodulin. These findings suggest that PMCA2 and 3 isoforms may be involved in developmental and differentiation processes. PMID- 15332123 TI - Isolation and characterization of a serine/threonine protein kinase SOS2 gene from Brassica napus. AB - A full-length cDNA of a new serine/threonine (Ser/Thr) protein kinase gene, designated as BnSOS2 (GenBank Acc. No.AY310413), was cloned from Brassica napus by rapid amplification of cDNA ends (RACE). The full-length cDNA of BnSOS2 was 1779 bp and contained a 1539-bp open reading frame encoding a protein of 512 amino acids. Homology analysis shows that BnSOS2 strongly resembles other Ser/Thr protein kinase genes, and that its putative protein belongs to a typical Ser/Thr kinase family. Northern blot analysis reveals that BnSOS2 is salt-inducible. Our results indicate that BnSOS2 is a new member of the plant SOS2 gene family, which may play an important role in salt tolerance of plants. PMID- 15332124 TI - Biglycan is internalized via a chlorpromazine-sensitive route. AB - The small leucine-rich proteoglycan biglycan (BGN) is abundantly expressed in mesenchymal tissues. Its expression level is related to the phenotypic differentiation of cells. A dysregulation in BGN expression occurs under several pathological conditions, including glomerulonephritis, mesothelioma, pancreatic cancer and a mouse model of osteoporosis. Since the extracellular concentration of BGN is regulated both by secretion and endocytosis, we performed mechanistic studies on BGN endocytosis in human skin fibroblasts in vitro, using inhibitors of different endocytic routes. Chlorpromazine, an inhibitor of the clathrin coated pit-pathway reduced endocytosis of BGN in human skin fibroblasts by 40%, and decreased degradation of BGN by 66% Filipin, an inhibitor of the caveolae pathway, and Tyrphostin AG 1478, a specific inhibitor of EGF-receptor phosphorylation that partially inhibits endocytosis of the structurally related proteoglycan decorin, had no influence on BGN internalization and degradation. Our data indicates that the classical clathrin-mediated endocytic pathway is a major route for the internalization of BGN. Based on the differential susceptibility to pharmacological inhibition, it appears that BGN endocytosis seems to be at least in part mechanistically different from decorin uptake. PMID- 15332125 TI - Protective effect of vaccination with DNA of the H. Pylori genomic library in experimentally infected mice. AB - Immunologically mediated protection against H. pylori infection is an attractive alternative to antibiotic treatment. We compared the efficacy of conventional protein vaccination with that of genetic vaccination against experimental infection with H. pylori in mice. For oral immunization, we used the recombinant peptide of an antigenic fragment of UreB (rUreB) or H. pylori-whole cell lysate antigens, and for genetic immunization, we used recombinant pcDNA and pSec plasmids inserted with the fragment of ureB or DNA of the H. pylori genomic library. Mice were challenged with the mouse stomach-adapted H. pylori Sidney Strain. The detection of gastric bacterial colonization was performed by real time PCR of a 26-kDa Helicobacter-specific gene, and the presence of serum H. pylori-specific antibodies was determined using direct ELISA assay. The most effective treatment appeared to be oral vaccination with rUreB and either intramuscular or intradermal vaccination with DNA of the H. pylori genomic library. Intradermal genetic vaccination with genomic library DNA significantly increased the IgG antibody response. Our study revealed acceptable efficacies of genetic vaccination with DNA of the H. pylori genomic library. PMID- 15332126 TI - The distribution of peripheral blood dendritic cells assayed by a new panel of anti-BDCA monoclonal antibodies in healthy representatives of the polish population. AB - A growing number of studies are being performed on the role of dendritic cells (DCs) in the etiopathogenesis of various conditions. Therefore, it is extremely important to establish the best comparable methods for the determination of the absolute count of blood dendritic cells (BDCs) or their subsets, and the reference normal values for comparisons. The aim of our study was to assess a normal profile of BDCs in the non-cultured human blood of healthy Polish volunteers. BDCs were detected among peripheral blood mononuclear cells (PBMC) from 99 healthy people, aged 18-56. Based on the panel of novel anti-BDCA1, BDCA2 and BDCA3 monoclonal antibodies (MoAbs), three main subpopulations of BDCs were distinguished: two myeloid types of BDCs, MDC1(BDCA-1+/ CD11c+ /HLA-DR+) or MDC2 (BDCA-3+/CD32-/CD64-/HLA-DR+), and a plasmacytoid subtype, PDC (BDCA 2+/CD123+/HLA-DR+). The number and percentage of BDCs were correlated with the age, gender, photosensitivity (phototype, minimal erythemal dose -- MED) and morphological parameters of the healthy volunteers. BDCs represented 0.83% of the PBMC and the median total BDC number was 44.0 cell/microl. The total BDC number correlated with the WBC count (rho=0.40, p=0.001) as well as with the lymphocyte and monocyte counts (rho=0.20, p=0.045 and rho=0.26, p=0.009, respectively). The median percentage of the MDC1 count (0.20%) was twice as high as the MDC2 count (0.10%). The median PDC count was 28.2 cell/microl, and these cells represented 0.50% of the PBMC. There was a positive correlation between PDC and skin photosensitivity (rho=0.28, p=0.005). An inverse correlation between the PDC count and the age of the examined volunteers was also found (rho=-0.22, p=0.029). Our study provides the first referential data on normal rates and counts of BDCs and their subpopulations, assessed by the new panel of anti-BDCA MoAbs, in healthy Polish subjects. The method used in the study allowed the determination of BDCs and their subset numbers in a relatively small blood volume. PMID- 15332127 TI - The effect of brachytherapy on antioxidant status and lipid peroxidation in patients with cancer of the uterine cervix. AB - The aim of this study was to investigate the effect of brachytherapy on lipid peroxidation and antioxidant status in patients with uterine cervix cancer. The study was conducted on 84 uterine cervix cancer patients from the Brachytherapy Department of the Regional Centre of Oncology in Bydgoszcz. Patients with uterine cervix cancer were found to have elevated levels of lipid peroxidation and antioxidant defence system impairment relative to healthy females. The results of the study indicate that brachytherapy has no direct effect on the antioxidant system of patients with uterine cervical carcinoma. However, the normalisation of catalase and glutathione peroxidase activity and erythrocyte TBARS level observed six months after the end of therapy may be due to the arrest of the progression of the disease. PMID- 15332128 TI - Genotoxicity of lead in lupin root cells as evaluated by the comet assay. AB - This paper presents the results of a study on the influence of lead (Pb(+2)) on DNA integrity on plant cells. The study was performed on the root tips of lupin (Lupinus luteus cv. Juno) seedlings treated with two selected concentrations of Pb(NO3)2: 150 and 350 mg l(-1), which were found to inhibit root growth by 50% and 70%, respectively [Rucinska et al. Plant Physiol. Biochem. 37 (1999) 37187 37194]. Roots exposed to those external lead concentrations took up about 50 and 70 mg l(-1) Pb(+2) g(-1) fresh weight (FW) over 48 h of incubation. A dose dependent increase in the degree of root injury was observed in the presence of both tested concentrations. The genotoxicity of lead in lupin root cells was analysed using a mild alkaline comet assay at pH 12.3, which allows the detection of single strand breaks. The quantity of the DNA fragments migrating away from the nuclear remnant (tail area) increased proportionally to the lead content inside the roots, and was positively correlated with the degree of root injury. At 150 mg l(-1) Pb(+2), a high frequency distribution of nuclei having large values of tail lengths and moments was observed. By contrast, the number of nuclei with minimum values of these parameters increased at 350 mg l(-1) Pb(+2). This data suggests that lead at low concentrations induces the formation of short, rapidly migrating DNA fragments, whereas at higher concentrations, lead probably causes other changes to DNA that result in slower DNA migration in the electric field. PMID- 15332129 TI - Extension of the Messapia x dicoccoides linkage map of Triticum turgidum (L.) Thell. AB - A set of recombinant inbred lines (RIL) derived from a cross between the cultivar Messapia of durum wheat (Triticum turgidum var. durum) and the accession MG4343 of T. turgidum var. dicoccoides was analysed to increase the number of assigned markers and the resolution of the previously constructed genetic linkage map. An updated map of the durum wheat genome consisting of 458 loci was constructed. These loci include 261 Restriction Fragment Length Polymorphisms (RFLPs), 91 microsatellites (Simple Sequence Repeats, SSRs), 87 Amplified Fragment Length Polymorphisms (AFLPs), two ribosomal genes, and nine biochemical (seven seed storage proteins and two isozymes) and eight morphological markers. The loci were mapped on all 14 chromosomes of the A and B genomes, and covered a total distance of 3038.4 cM with an average distance of 6.7 cM between adjacent markers. The molecular markers were evenly distributed between the A and the B genomes (240 and 218 markers, respectively). An additional forty loci (8.8%) could not be assigned to a specific linkage group. A fraction (16.4%) of the markers significantly deviated from the expected Mendelian ratios; clusters of loci showing distorted segregation were found on the 1B, 2A, 2B, 3A, 4A, 7A and 7B chromosomes. The genetic lengths of the chromosomes range from 148.8 cM (chromosome 6B) to 318.0 cM (chromosome 2B) and approximately concur with their physical lengths. Chromosome 2B has the largest number of markers (47), while the chromosomes with the fewest markers are 3A and 6B (23). There are two gaps larger than 40 cM on chromosomes 2A and 3B. The durum wheat map was compared with the published maps of bread and durum wheats; the order of most common RFLP and SSR markers on the 14 chromosomes of the A and B genomes were nearly identical. A core-map can be extracted from the high-density Messapia x dicoccoides map and a subset of uniformly distributed markers can be used to detect and map quantitative trait loci. PMID- 15332130 TI - DNA methylation polymorphism in a set of elite rice cultivars and its possible contribution to inter-cultivar differential gene expression. AB - RAPD (randomly amplified polymorphic DNA) and ISSR (inter-simple sequence repeat) fingerprinting on HpaII/MspI-digested genomic DNA of nine elite japonica rice cultivars implies inter-cultivar DNA methylation polymorphism. Using both DNA fragments isolated from RAPD or ISSR gels and selected low-copy sequences as probes, methylation-sensitive Southern blot analysis confirms the existence of extensive DNA methylation polymorphism in both genes and DNA repeats among the rice cultivars. The cultivar-specific methylation patterns are stably maintained, and can be used as reliable molecular markers. Transcriptional analysis of four selected sequences (RdRP, AC9, HSP90 and MMR) on leaves and roots from normal and 5-azacytidine-treated seedlings of three representative cultivars shows an association between the transcriptional activity of one of the genes, the mismatch repair (MMR) gene, and its CG methylation patterns. PMID- 15332131 TI - Isolation, cloning and characterisation of motifs containing (GA/TC)n repeats isolated from vetch, Vicia bithynica. AB - Microsatellites are widely distributed in plant genomes and comprise unstable regions that undergo mutational changes at rates much greater than that observed for non-repetitive sequences. They demonstrate intrinsic genetic instability, manifested as frequent length changes due to insertions or deletions of repeat units. Detailed analysis of 1600 clones containing genomic sequences of Vicia bithynica revealed the presence of microsatellite repeats in its genome. Based on the screening of a partial DNA library of plasmids, 13 clones harbouring (GA/TC)n tracts of various lengths of repeated motif were identified for further analysis of their internal sequence organization. Sequence analyses revealed the precise length, number of repeats, interruptions within tracts, as well as sequence composition flanking the repeat motifs. Representative plasmids containing different lengths of (GA/TC)n embedded in their original flanking sequence were used to investigate the genetic stability of the repeats. In the study presented herein, we employed a well characterised and tractable bacterial genetic system. Recultivations of Escherichia coli harbouring plasmids containing (GA/TC)n inserts demonstrated that the genetic instability of (GA/TC)n microsatellites depends highly on their length (number of repeats). These observations are in agreement with similar studies performed on repetitive sequences from humans and other organisms. PMID- 15332133 TI - Six-month CTS update report. PMID- 15332132 TI - First Nations pneumonia admissions: different patients or different attitudes? PMID- 15332135 TI - Hospitalization for community-acquired pneumonia in Alberta First Nations Aboriginals compared with non-First Nations Albertans. AB - BACKGROUND: The rates and outcomes of hospital admission for community-acquired pneumonia between First Nations Aboriginal and non-First Nations groups were compared. METHODS: Alberta administrative hospital abstracts from April 1, 1997, to March 31, 1999, were analyzed, and each case of a First Nations Aboriginal person with pneumonia was matched by age and sex with three non-First Nations persons with pneumonia. RESULTS: The First Nations Aboriginal age and sex adjusted hospital discharge rate was 22 per 1000 (95% CI 20.7 to 23.6) compared with 4.4 per 1000 (95% CI 4.4 to 4.5) for the general population of Alberta. After accounting for comorbidity and severity of pneumonia, in-hospital mortality and hospital length of stay were lower for First Nations Aboriginals compared with the matched non-First Nations group (odds ratio 0.49; 95% CI 0.37 to 0.66, and odds ratio 0.87; 95% CI 0.79 to 0.97, respectively). The odds for 30-day hospital readmission were higher in First Nations Aboriginals compared with the non-First Nations group (odds ratio 1.42; 95% CI 1.21 to 1.68). The cost per hospital admission for First Nations Aboriginals was 94% of the average cost for the matched non-First Nations group (CDN4,206 dollars). However, their median daily cost was 1.25 times higher (95% CI 1.14 to 1.36) than the matched non-First Nations group. CONCLUSIONS: First Nations Aboriginals had higher rates of hospitalization, rehospitalization and hospital costs for community-acquired pneumonia than non-First Nations Albertans. It was unlikely that the high rate of hospitalizations in First Nations Aboriginals was due to more severe pneumonia or greater comorbidity. Other unexplained factors increase the burden of this disease in First Nation Aboriginals. PMID- 15332136 TI - Asthma and allergic rhinitis in Quebec children. AB - BACKGROUND: The Health and Social Survey of Quebec Children and Youth, conducted on representative samples of children nine, 13 and 16 years of age, provided data on the prevalence and determinants of asthma and allergic rhinitis in Quebec. OBJECTIVES: To determine the prevalence of asthma and allergic rhinitis among children in the province of Quebec and to identify the determinants of these pathologies. METHODS: Three groups of more than 1100 children aged nine, 13 and 16 years were recruited. Respiratory symptoms were documented using the International Study of Asthma and Allergies in Childhood questionnaire. Questions enquiring about family income, smoking, degree of urbanization of the child's school's location and various variables related to indoor air were also included. The comparisons of proportions were done using the chi2 test. RESULTS: The prevalence rates for reported history of asthma varied from 14% to 15% depending on the age group. The prevalence of wheezing in the past year was 7% to 8%. Asthma was the primary cause of the limitation of activities due to a health problem in nine- and 13-year-old Quebecers, and the second most common cause in 16-year-old Quebecers. The prevalence of rhinitis, rhinoconjunctivitis and reported history of hay fever increased with age, reaching 28.0%, 15.9% and 21.1%, respectively, in the 16-year-old group. The prevalence of asthma and wheezing was associated with family history and allergies, and inversely related to family income. CONCLUSIONS: The prevalence of childhood asthma is high in the province of Quebec. It is a major cause of the limitation of activities due to a health problem for young Quebecers. A family history of asthma and an atopic predisposition are important determinants in the development of asthma in Quebec. PMID- 15332137 TI - The costs of implementing the 1999 Canadian Asthma Consensus Guidelines recommendation of asthma education and spirometry for the family physician. AB - BACKGROUND: National and international asthma guidelines recommend that patients with asthma be provided with asthma education and spirometry as a component of enhanced asthma care. The cost of implementing these interventions in family physician practices is not known. OBJECTIVE: The objective of the present study was to determine the cost of providing recommended asthma care to adult patients in the family practice setting. METHODS: The present study was conducted using three scenarios of care in family practice. Small, medium and large asthmatic patient populations were used. The incremental costs of implementing enhanced asthma care based on the Canadian Asthma Consensus Guidelines, including the provision of spirometry and asthma education in both group and individual sessions, and the resources required for these interventions were calculated for each scenario. RESULTS: For a physician with 50 asthmatic patients, the cost of providing enhanced asthma care with spirometry and group education sessions was approximately 78 dollars per patient in the first year of implementation. For individual sessions, the cost increased to 100 dollars per patient for the first year. If the physician had 100 asthmatic patients, the per patient cost would decrease; however, the overall cost of the program would be 7,000 dollars. CONCLUSIONS: The costs of providing enhanced asthma care are significant. In most cases, physicians are inadequately reimbursed (or not reimbursed) for these interventions. In light of the evidence of the effectiveness of these interventions, health insurance plans should consider adding these services to fee schedules. PMID- 15332138 TI - West Nile virus infection in the intensive care unit: a case series and literature review. AB - BACKGROUND: West Nile virus (WNV) is a rapidly spreading infectious disease in North America. Critical care issues related to WNV are not well described. OBJECTIVES: Three cases of severe WNV meningoencephalitis with flaccid paralysis are reported and relevant critical care issues are highlighted. METHODS: Case series and a review of the literature. RESULTS: Three patients with WNV meningoencephalitis and flaccid paralysis were admitted to the authors' academic, tertiary-care intensive care unit (ICU) in the late summer of 2002. All three patients were middle-aged or elderly and presented with a febrile illness that preceded or coincided with their neurological symptoms. Confirmation of WNV infection was problematic because each patient had at least one initial negative serum serology test. Radiological testing yielded nonspecific results. Serial electroencephalograms were consistent with severe toxic metabolic encephalopathy in all cases. All patients had a severe, diffuse axonal polyneuropathy demonstrated by nerve conduction studies and electromyography. Prolonged mechanical ventilation resulted in ICU lengths of stay of 44 to 118 days. At one point, 21% of the ICU beds were dedicated for these patients. All three patients died in hospital - two following the withdrawal of life support. One patient demonstrated resolving encephalitis and was discharged from the ICU after a 118 day ICU stay, but later died in a step-down unit. CONCLUSIONS: The management of WNV-related critical illness creates challenges in making a timely and accurate diagnosis, and predicting patient morbidity and mortality. As a consequence, end of-life discussions with families are especially difficult. The prolonged ICU length of stay and growing incidence of this disease may challenge limited critical care resources. PMID- 15332139 TI - Asbestosis and probable microscopic polyangiitis. AB - Several inorganic dust lung diseases (pneumoconioses) are associated with autoimmune diseases. Although autoimmune serological abnormalities are common in asbestosis, clinical autoimmune/collagen vascular diseases are not commonly reported. A case of pulmonary asbestosis complicated by perinuclear antineutrophil cytoplasmic antibody (myeloperoxidase) positive probable microscopic polyangiitis (glomerulonephritis, pericarditis, alveolitis, multineuritis multiplex) is described and the possible immunological mechanisms whereby asbestosis fibres might be relevant in induction of antineutrophil cytoplasmic antibodies are reviewed in the present report. PMID- 15332142 TI - The second labour of Martin. PMID- 15332140 TI - Massive pleural effusion in an 18-year-old girl with Ewing sarcoma. AB - Ewing sarcoma is a bone tumour that commonly appears between ages five and 10 in the diaphysis of the long bones and predominantly presents with pain and swelling. The case of an 18-year-old girl who presented with back pain, cough, dyspnea, weakness and fever is described. Chest radiograph showed a homogenous density in the middle and inferior zones of the right hemithorax. Thoracic computed tomography revealed a diffuse pleural effusion and a 6.99 cm x 4.45 cm solid mass composed of lobulated, small cystic lesions and calcifications in the right hemithorax. Biochemical analysis of pleural fluid showed hemorrhagic effusion and exudate. A pleural needle biopsy demonstrated solid uniform tumour cells with narrowed cytoplasm, round nuclei and uncertain nucleoli. All of the tumour cell cytoplasms stained with CD99. The pathological examination supported Ewing sarcoma. Three-phase Tc-99m methylene diphosphonate scintigraphy of the whole body showed pathological tracer uptake in a broad area of the eighth costal bone and in smaller areas of the ninth and 10th costal bones. This case is reported because Ewing sarcoma is a rare cause of pleural effusion in clinical practice among younger adults. PMID- 15332143 TI - The Ontario Cardiac Rehabilitation Pilot Project. AB - In February 2001, the Ontario Ministry of Health and Long-Term Care announced a $9.6 million, 15-month pilot project (the Pilot) to implement and evaluate a comprehensive, multifactoral model of cardiac rehabilitation (CR) service delivery at 17 sites across Ontario. This is the second paper in a three-part, policy-related series which provides a summary of the Ontario CR Pilot model and the Pilot implementation and evaluation methodology. The aim of the present paper was to outline the goals of the Pilot, the Pilot model of care, the organizational structure that facilitated implementation of the model, and the operational procedures that were put in place to evaluate patient outcomes and the generalizability of a regional model of CR service delivery. The model was based on the findings and recommendations of the Cardiac Care Network of Ontario's 1999 Consensus Panel on Cardiac Rehabilitation and Secondary Prevention, which was described in part one of this series. An upcoming final paper will describe the outcomes of the project and its recommendations for CR health policy decisions in Ontario. PMID- 15332144 TI - Management of heart failure in Canadian long-term care facilities. AB - BACKGROUND: In long-term care (LTC) facilities, heart failure is common but undertreated. No Canadian studies of heart failure in LTC facilities have been reported. OBJECTIVES: To estimate the prevalence of heart failure in Canadian LTC facilities; to document the management of heart failure in LTC; and to describe characteristics of LTC residents with heart failure and predictors of adherence to treatment guidelines. METHODS: Cross-sectional survey in eight LTC facilities lodging 1223 residents. RESULTS: The prevalence of heart failure was 20%. LTC residents with heart failure were older, more often women, and more functionally impaired and burdened by comorbidity than were participants in heart failure trials. Documentation supporting the heart failure diagnosis was inadequate, with some symptoms possibly misattributed to chronic obstructive pulmonary disease. Angiotensin-converting enzyme (ACE) inhibitors were prescribed to 55% of residents, although only 45% received appropriate doses. Residents with hypertension or diabetes mellitus, using nitrates or who were male were more likely to receive ACE inhibitors. Appropriate ACE inhibitor doses were associated with functional impairment, nitrate use and recent hospitalization. Documentation of systolic dysfunction was associated with a greater likelihood of ACE inhibitor use. Beta-blockers were prescribed to 25% of residents, who were more likely to be using nitrates, have ischemic heart disease or had been recently hospitalized, and less likely to have chronic obstructive pulmonary disease. Residents with atrial fibrillation were more likely to be prescribed digoxin. Potentially hazardous regimens were prescribed to 43% of residents. CONCLUSIONS: Heart failure is common in Canadian LTC facilities. Management of heart failure in LTC does not conform to guidelines. Improved diagnostic methods tailored for frail elderly patients must be developed. Studies are needed to understand and identify factors influencing prescribing for heart failure medication in LTC. PMID- 15332145 TI - Cardiac hemochromatosis in an HFE His63Asp (187C->G) heterozygote. PMID- 15332146 TI - CCAA are diagnosable antemortem. PMID- 15332148 TI - ACC/AHA guidelines for the management of patients with ST-elevation myocardial infarction--executive summary: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Writing Committee to Revise the 1999 Guidelines for the Management of Patients With Acute Myocardial Infarction). PMID- 15332174 TI - From molds and macrophages to mevalonate: a decade of progress in understanding the molecular mode of action of bisphosphonates. AB - Although bisphosphonates were first used as therapeutic agents to inhibit bone resorption in the early 1970s, their mode of action at the molecular level has only become fully clear within the last few years. One of the reasons for this lack of understanding was the difficulty in isolating large numbers of pure osteoclasts for biochemical studies. In the last decade, the identification of appropriate surrogate models that reflected the antiresorptive potencies of bisphosphonates, such as Dictyostelium slime molds and macrophages, helped overcome this problem and proved to be instrumental in elucidating the molecular pathways by which these compounds inhibit osteoclast-mediated bone resorption. This brief review summarizes our current understanding of these pathways. PMID- 15332176 TI - Fixed partial denture or single-tooth implant restoration? Statistical considerations for sequencing and treatment. AB - The choice to replace a single missing tooth can be based on the primary decision that the restorability of the tooth is in doubt. Many teeth are decimated by incipient or recurrent caries, trauma, endodontic complications, or periodontal disease which requires extraction. It is our objective to familiarize the participant with literature comparing success rates of fixed partial dentures and single-tooth implant restorations and a repertoire of prosthodontic techniques used in replacement of single missing teeth with osseointegrated dental implants. The fixed partial denture (FPD) has been regarded as the standard of care for some time in replacement of single and multiple missing teeth. Many studies surveying long term survival have been compiled and analyzed to arrive at a generalized outcome. Most of these studies arrive at common conclusions. Studies surveying success of single-tooth implant-supported restorations are not comparably abundant nor survey for comparable time as those for FPDs. Although, many of the outcomes are statistical survival estimates such as Kaplan-Meier survival tables, implant restorations in partially dentate patients are a predictable means of tooth replacement. There are certain factors which make FPD more appropriate and conversely factors which make an implant restoration more appropriate. Indications and contraindications for each treatment scenario will also be reviewed based on the literature and clinical experience. It is hoped that the practitioner will be able to appropriately identify those cases in which either an FPD or an implant restoration is the appropriate treatment option. PMID- 15332177 TI - The OSU frame: a novel approach to fabricating immediate load fixed-detachable prostheses. AB - PURPOSE: Interest in the concept of immediate or early loading of dental implants is increasing. The survival of these implants, particularly in the anterior mandible, has been demonstrated in a variety of clinical reports for different implant systems. However, to make immediate loading a reality for the patient, the restorative dentist must coordinate care with the surgeon and the laboratory fabricating the prosthesis. This article describes a method that allows restoration of the edentulous mandible in 2 days. MATERIALS AND METHODS: Five implants were placed in the mandible and restored in 2 days with a final fixed detachable prosthesis that was fabricated using the OSU Frame. There is little change to the surgical protocol whether the implants are loaded immediately or loading was delayed. The step-by-step technique is described. RESULTS AND CONCLUSION: There are several benefits to this method over those currently available. No changes in surgical armamentarium technique are required. The technique may be adapted to any implant system and it offers improved flexibility over the existing systems. The OSU Frame accommodates variability in arch form and suboptimal implant placement, facilitating immediate implant loading with the final prosthesis for nearly any edentulous implant patient. PMID- 15332178 TI - A clinical evaluation of the Zygoma fixture: one year of follow-up at 16 clinics. AB - PURPOSE: To evaluate treatment outcome with Zygoma fixtures (Nobel Biocare, Goteborg, Sweden) with regard to fixture survival, patient satisfaction, and function of prosthesis replacement. MATERIALS AND METHODS: The treatment outcome of 76 patients treated with 145 Zygoma fixtures at 16 centers was evaluated. Patient's and dentist's evaluations of the functional and aesthetic outcome of the treatment were assessed at delivery of prosthesis and at the 1-year follow-up visit. At the 1-year follow-up visit, the status of the peri-implant mucosa around the abutments and the amount of plaque were registered. RESULTS: Sixty-six of the 76 patients, with 124 Zygoma fixtures supporting the prosthetic restorations, were evaluated at the 1-year follow-up. The overall survival rate for the Zygoma fixtures was 97.9% after 1-year of follow-up. Eighty percent of the patients were fully satisfied with both aesthetic and functional outcome at the time of prosthetic insertion and at the 1-year follow-up. All reported data from the dentists, with the exception of one restoration with several abutment screw loosenings, scored from acceptable to excellent for the aesthetic and functional outcome of the treatment. The status of peri-implant mucosa was recorded as normal in approximately 60% of the sites. Plaque, when present, was more often detected on the palatal surfaces compared with the buccal surfaces. CONCLUSION: This 1-year follow-up of Zygoma fixtures has shown good results with an acceptable number of minor complications and a majority of satisfied patients. PMID- 15332179 TI - Immediate loading of the edentulous mandible: delivery of the final restoration or a provisional restoration--which method to use? AB - PURPOSE: Edentulous patients desire restoration of their chewing ability as soon as possible after placement of dental implants. The purpose of this article is to provide clinicians with evidence that immediate loading of implants placed into the anterior mandible can predictably provide the patient with early functional rehabilitation. Two methods are presented that illustrate delivery of the final restoration or a provisional implant-borne prosthesis immediately after implant placement. MATERIALS AND METHODS: A literature search was performed which produced 14 articles in the English literature that provided sufficient evidence that immediate loading is now not experimental and can be recommended as an acceptable treatment alternative. RESULTS: Two practical methods to achieve immediate function are presented in a step-wise manner to illustrate how to deliver this service to the patient. CONCLUSION: Based on our literature review, immediate loading of the edentulous mandible with an implant-borne restoration is an acceptable and predictable method to deliver efficient return of function for the edentulous patient. PMID- 15332180 TI - Interactive imaging for implant planning, placement, and prosthesis construction. AB - PURPOSE: This review describes a new interactive imaging program that allows computed tomography (CT) images to be used to virtually place dental implants and construct a precise guide splint and final prosthesis for delivery at the time of implant placement. MATERIALS AND METHODS: Patients with edentulous arches were recruited to participate in a trial program to assess the feasibility of using CT images in a 3-dimensional image-based program (Oralim; Medicine NV, Belgium) for planning and placing dental implants. Patients meeting the criteria received a CT of the appropriate arch using a denture with radiopaque markers indexed to the opposing arch. Acquisition slices of 0.6 mm or less was required. The software allowed precise planning for implant placement after which the planned case was sent to a manufacturing facility for splint and prosthesis construction. RESULTS: The guide splint and final prosthesis were returned to the clinical site for implant placement. An implant map was provided for each patient showing the diameter and length of the implant and abutments to be used in each predetermined implant site. The surgery was accomplished without a flap, and the prosthesis was delivered immediately. Occlusion required only minor adjustments in all cases, and immediate function was accomplished. CONCLUSION: Interactive computer imaging can allow precise planning for implant position and the images can be used for guide splint and final prosthetic fabrication prior to surgery. This technology is powerful, easy to use, and is a significant advance in implant dentistry. PMID- 15332181 TI - Maxillary lateral incisor implants: planning with the aid of orthodontics. AB - Implants are commonly used to replace congenitally missing lateral incisors in adolescent orthodontic patients. However, these restorations are often challenging for the orthodontist, surgeon, and restorative dentist. In some patients the space across the alveolar crest is too narrow to permit the surgeon to place the implant. Occasionally the root apices of the adjacent central incisor and canine are in close proximity. In other cases the ridge thickness could be inadequate and require augmentation. When the orthodontist opens the space, the papilla heights are adversely affected. Some adolescent patients have altered passive eruption after orthodontic treatment, which affects the level of the gingival margins. Finally, questions commonly arise regarding the appropriate age for implant placement in these young patients. If not addressed, these issues could compromise the aesthetics of the implant restoration. This article will use several clinical examples to discuss each of these 6 potential problems and provide guidelines for their solutions. PMID- 15332182 TI - Single-stage Straumann dental implants in the aesthetic zone: considerations and treatment procedures. AB - The aim of this paper was to identify factors significant to the use of single stage dental implants in the aesthetic zone and discuss treatment considerations related to them. In view of available literature and clinical experiences, treatment outcomes associated with the use of single-stage Straumann implants (Institut Straumann AG, Waldenburg, Switzerland) in the aesethetic zone can be considered positive and predictable. PMID- 15332183 TI - Horizontal ridge augmentation using human mineralized particulate bone: preliminary results. AB - PURPOSE: We evaluated the hypothesis that particulate human mineralized bone can be used to augment the thin mandibular ridge followed by implant placement 4 months after augmentation. The thin ridge prevents implant placement without grafting. PATIENTS AND METHODS: Eleven consecutive patients with thin posterior mandibular ridges had approximately 1 cc of particulate human mineralized bone placed through a tunneling approach to augment the ridge width. Thirty-five implants were placed into 13 ridges after graft consolidation, with 1 implant failure. RESULTS: All grafted ridges were able to have at least a 3.25 mm diameter implant placed. One implant failed to integrate. One-year follow-up indicates stability of the augmentation. CONCLUSION: This preliminary report indicates the potential for this technique to substitute for more invasive procedures. PMID- 15332184 TI - Single-implant restorations: a contemporary approach for achieving a predictable outcome. AB - The replacement and restoration of missing single teeth with dental implants are routine procedures. However, because of the relatively short clinical history of this procedure in dentistry, custom-tailored protocols for this procedure do not exist. This article will outline the various surgical and restorative steps involved in the treatment planning, fabrication, and delivery of single-implant restoration, and will critically analyze the existing clinical concepts and provide recommendations based on a contemporary approach. As data for and experience with this procedure increase the same general guidelines for all types of implant restorations may not apply. Emphasis will be placed on differentiating between the various types of single-implant restorations, comparing and contrasting the differences between the different types of single-implant restorations, and revising some of the existing concepts of this treatment modality. PMID- 15332185 TI - Reconstruction of extensive maxillary defects using zygomaticus implants. AB - PURPOSE: Maxillary reconstruction after maxillectomy remains a great challenge for the reconstructive oral and maxillofacial surgeon. This article is a clinical retrospective analysis of patients reconstructed with zygomaticus implants after maxillary ablation. PATIENTS AND METHODS: The design of the study was a retrospective review of 9 patients requiring near-total or total maxillectomy for pathologic reasons. Clinical records, photographs, and radiographs were studied. Financial billing statements were reviewed to determine charges for implant reconstruction and method of payment. RESULTS: Maxillary reconstruction using zygomaticus and standard endosseous implants was performed in 9 patients. Maxillary resection was performed for the following reasons: salivary gland malignancy (n = 2), squamous cell carcinoma (n = 5), maxillary mucormycosis (n = 1), and extensive maxillary atrophy and infection secondary to subperiosteal maxillary implant placement (n = 1). A total of 28 zygomaticus implants and 10 standard endosseous implants were used to reconstruct the 9 patients. Six zygomaticus implants and 3 standard endosseous implants failed. The time of zygomaticus implant placement ranged from placement at the time of resection to 3.2 years after the resection. Five patients received radiation therapy. Five patients have been reconstructed with a maxillary obturator and have been functioning with the prosthesis for a minimum of 2 years. CONCLUSION: The combination of zygomaticus and standard endosseous implants can be used to reliably reconstruct patients after extensive resection of the maxilla. PMID- 15332186 TI - Strategies for management of single-tooth extraction sites in aesthetic implant therapy. AB - Achieving predictable aesthetic outcomes following the planned removal of a natural tooth suffering from structural, endodontic, or periodontal compromise depends on a multitude of factors that, once identified, guide the implant team in selecting the surgical and prosthetic treatment options best suited for the individual clinical scenario. This article presents pertinent information regarding systematic patient evaluation and special treatment planning considerations for patients facing the loss of a single tooth in an area of high aesthetic importance. The rationale for and details of performing the Bio-Col site preservation technique at the time of tooth removal; guidelines for immediate versus delayed implant placement; selection and sequencing of site development procedures according to the types of alveolar ridge defects encountered following tooth removal despite the use of site preservation; and a description of prosthetic techniques necessary for the successful management of single-tooth extraction sites in areas of aesthetic concern also are reviewed. PMID- 15332187 TI - Immediate loading of definitive implants in the edentulous mandible using a fixed provisional prosthesis: The denture conversion technique. AB - PURPOSE: This article presents a clinical technique for immediate implant loading in the completely edentulous mandible. MATERIALS AND METHODS: The protocol recommends the placement of 4 to 5 root-form implants in the anterior mandible between the mental foramina. Upon insertion the implants must have favorable primary stability. Abutments are added to the implants and the patient's lower denture is converted into an immediate load provisional fixed prosthesis. RESULTS: The denture conversion technique has been shown to provide predictable results for immediate implant loading in the edentulous mandible. The final prosthesis is fabricated upon integration of the implants in 3 months. CONCLUSION: The denture conversion technique offers several advantages because it may be used with most commercially available implant systems and it incorporates conventional implant components. Guidelines for patient selection, presurgical preparation, implant surgery, prosthesis fabrication, and postoperative care are reviewed. PMID- 15332188 TI - Navigation surgery for dental implants: assessment of accuracy of the image guided implantology system. AB - PURPOSE: The purpose of this study is to assess the accuracy of the navigation provided by the Image Guided Implantology system (DenX Advanced Dental Systems, Moshav Ora, Israel), which was designed to guide the surgeon in the placement of dental implants. MATERIALS AND METHODS: Seven jaw models incorporating special ceramic spheres were imaged by dental computerized tomography. The coordinates of these reference ceramic markers were calculated by the Image Guided Implantology and compared with their actual position coordinates as identified on the computed tomography image. RESULTS: The overall mean spatial navigation error was 0.35 +/- 0.14 mm. The 1-tailed probability of any single measurement exceeding 0.75 mm was less than 0.003, and that exceeding 1 mm was less than 0.0001. CONCLUSION: The Image Guided Implantology system provides highly accurate navigation with less than 0.73 mm error, which is acceptable in dental implantology. The accurate reporting of the exact position of the drilling bur should minimize the potential risk of damage to critical anatomic structures. The accurate intraoperative navigation allows the surgeon to precisely transfer the presurgical plan to the patient. PMID- 15332189 TI - Technique for in-office cranial bone harvesting. AB - PURPOSE: This is a report of a technique of cranial bone harvesting suitable for the outpatient setting. MATERIALS AND METHODS: Bone scrapers are used for the harvesting of cranial bone shavings with the patient under intravenous sedation or general anesthesia. RESULTS: Graft volumes larger than that usually obtainable from intraoral sites and the tibia have been harvested utilizing this technique. In a series of 8 first patients, the largest volume of bone obtained was 14 cc with no complications related to the donor sites. These cases include the following types of pre-implant reconstructive procedures: large unilateral sinus grafting, bilateral sinus grafting/guided-bone regeneration of an entire alveolar ridge, inlay grafting of the alveolus, inlay grafting in association with distraction osteogenesis, subnasal grafting, alveolar cleft grafting, closure of large oroantral defects combined with sinus grafting, and grafting of an grossly atrophic mandible with simultaneous placement of dental implants via the submental approach. CONCLUSION: This is a safe bone harvesting technique providing an alternative source of autogenous bone graft. PMID- 15332190 TI - Distraction osteogenesis of the nasal and antral osseous floor to enhance alveolar height. AB - PURPOSE: The purpose of the study was to obtain information on the possibility of using distraction osteogenesis (DO) to increase the height of posterior atrophic maxillary alveolar ridges by distracting very small alveolar crest bone segments and, secondly, to determine if it is possible to distract the bony antral floor itself, producing new bone between the sinus membrane and the alveolar crest. MATERIALS AND METHODS: Three adult Papio anubis baboons were rendered edentulous in the posterior maxilla. Bilateral osteotomies were performed to produce a thin margin of crestal alveolar bone and in one area, a complete exposure of the antral floor. Distractors were placed against the antral and nasal floors and the thin 1- x 1.5-mm thick alveolar bone crest was distracted 10 mm over a period of 10 days. Animals were labeled with tetracycline at 3 and 4 months and were sacrificed at 5 months. RESULTS: A complete osseous regeneration of the nasal floor and alveolar ridge greater than 10 mm in height was produced in all cases. Histomorphometric analysis of the DO-induced osseous areas revealed cancellous bone with increasing thickness during the third and fourth month and formation of new bone cortices increasing in thickness during the same time. Significant new bone also formed in the antral floor areas that had been purposely interdicted by the osteotomies. CONCLUSION: The results of this study indicate that very small segments of bone of the posterior maxilla can be distracted to produce significant increases in the alveolar bone height and that a new osseous nasal antral floor enhancement can be produced by this technique. PMID- 15332191 TI - Bone healing after the inferior alveolar nerve lateralization: a histologic study in rabbits (Oryctolagus cuniculus). AB - PURPOSE: The purpose of this article is to evaluate the bone healing around 2 different dental implant surfaces after the lateralization surgery of the inferior alveolar nerve (IAN) during an 8-week healing period in rabbits, and to check if there is any interaction between the implants and the nerve. MATERIALS AND METHODS: The IAN lateralization was performed in an experimental rabbit model. Eight adult female rabbits (Oryctolagus cuniculus) underwent the surgical procedure, and 1 implant was placed on each side of the mandible while the nerve was lateralized. On both sides, the nerve was repositioned directly in contact with the implant surface. With the intention of obtaining comparative results, smooth titanium implants were installed on the right side, and blasted Al2O3 ones were placed on the left. During the healing period, bone tracers were administered subcutaneously to check different periods of bone ingrowth. RESULTS: Histologic section, regardless of the studied surface, showed bone remodeling around the nerve, without contact between the nerve and the implanted surface. The bone blocks containing implants were histomorphometrically examined through computerized analysis, and the results obtained showed that the bone remodeling around the nerve occurred during the first weeks of healing. Through analysis of variance, the blasted Al2O3 implants showed at least 2.5-fold greater bone neoformation compared with the smooth titanium implants. CONCLUSIONS: Our results showed that there was no significant difference in the healing process concerning the nerve between the 2 studied surfaces. No interaction was detected between the nerve and the implants. PMID- 15332192 TI - The "FAD" (Floating Alveolar Device): a bidirectional distraction system for distraction osteogenesis of the alveolar process. AB - PURPOSE: The following study illustrates preoperative and perioperative vector management in alveolar distraction using a new distraction system--the "Floating Alveolar Device" (FAD). The FAD is a bidirectional alveolar distractor that allows augmentation of an atrophic alveolar process in several planes, assures easy intraoperative positioning of the planned vector of distraction, and provides correction of the horizontal position of the transported segment during and after vertical distraction. PATIENTS AND METHODS: The FAD is composed entirely of stainless steel and has the following basic components: an upper member, a distraction rod, a lower base plate supporting the vertical force of the distraction rod, a jointed hinge that connects the upper and the lower members, and a tightening rod that provides blocking of the hinge. The clinician can manipulate and adjust the tightening rods, allowing a change in the angle of the hinge, thereby altering the transverse dimension of the vector of distraction. A total of 4 patients aged to 19 to 40 years underwent bidirectional alveolar distraction. All procedures were performed in the mandible. RESULTS: In all treated patients, planned distraction height and direction were achieved. In all cases it was possible to place implants at the planned time. CONCLUSIONS: The most common complication, axial displacement, is easily eliminated by moving the bone with the "floating" rod of the FAD during or immediately after the distraction period, according to the principles of the floating bone concept. PMID- 15332193 TI - Persistence and stability of delusions over time. AB - Traditional descriptions of delusions have emphasized the conviction with which they are held and their resistance to change. This study utilizes data from a large cohort of delusional subjects to assess the persistence and stability of delusional beliefs, and the predictors of change. Data were collected from 1,136 acutely hospitalized psychiatric patients, reinterviewed at 10-week intervals for 1 year. Persistence of delusional beliefs was determined for those delusional subjects with at least one follow-up visit (n = 405), and stability for the subset with delusions at two or more points in time (n = 262). Marked plasticity in delusional beliefs was observed, with one third of delusional subjects at any interview no longer delusional 10 weeks later. Persistence of delusions was associated with schizophrenia, global psychopathology, and having acted on a delusion, among other variables. Most subjects showed variation in the content of their primary delusion over time. Delusions appear to be more fluid over relatively short periods of time than has been suggested by many classic descriptions and contemporary formulations. PMID- 15332194 TI - DSM-IV obsessive-compulsive personality disorder: prevalence in patients with anxiety disorders and in healthy comparison subjects. AB - The relationship between obsessive-compulsive disorder (OCD) and obsessive compulsive personality disorder (OCPD) has not yet been fully clarified. The aim of the present study was to analyze DSM-IV OCPD prevalence rates in OCD and panic disorder (PD) patients to test for the specificity of the OCPD-OCD link, and to compare them to OCPD prevalence in a control group of subjects without any psychiatric disorder. A total of 109 patients with a principal diagnosis of DSM IV (SCID-I) OCD and 82 with PD were interviewed using the Structured Clinical Interview for DSM-IV Axis II Disorders (SCID-II) in order to assess the prevalence of OCPD. All patients with a coexisting axis I diagnosis were excluded from the study to eliminate confounding factors when evaluating the association between prevalence rates of OCPD and anxiety disorder diagnoses. An exclusion criteria was also a Hamilton Depression Rating Scale (HAM-D) score >/=16. A sample of comparison subjects (age 18 to 65 years) without any psychiatric disorder was recruited from people registered with two general practitioners (GPs), whether or not they consulted the doctor, in order to evaluate OCPD prevalence rate in the community. A significant difference was found between the prevalence of OCPD in OCD (22.9%) and in PD (17.1%) on one hand, and that in the comparison sample (3.0%) on the other. No differences were found between the two psychiatric groups, even when splitting the samples according to gender. Our study failed to support the hypothesis of a specific relationship between OCPD and OCD; we confirmed the higher prevalence rate of this personality disorder in OCD subjects with regard to the general population, but we also confirmed the higher rate of OCPD in another anxiety disorder which is phenomenologically well characterized and different from OCD, such as PD. PMID- 15332195 TI - Seasonal differences in psychopathology of male suicide completers. AB - Suicide is known to vary according to season, with peaks in the spring and troughs in the winter. The presence of psychopathology is a significant predictor of suicidality, and it is possible that the seasonal variation of suicide completion may be related to seasonality in the manifestation of psychiatric disorders common to suicide completers. In the current study, we evaluated 115 French-Canadian male suicide completers from the Greater Montreal Area for DSM-IV psychiatric disorders using proxy-based diagnostic interviews. Subjects were assessed for seasonal differences in the prevalence of DSM-IV psychiatric diagnoses just before their deaths. Diagnoses of major depressive disorder (MDD) without comorbid cluster B personality disorders, and schizophrenia were differently distributed between seasons. Most (63.4%) subjects with MDD committed suicide in the spring/summer (P =.038). However, closer examination revealed that depressed suicides with comorbid cluster B personality disorders did not show seasonality, while 83.3% of depressed suicides without comorbid cluster B personality disorders committed suicide in the spring/summer (P =.019). 87.5% of those suicides with schizophrenia committed suicide in the fall/winter (P =.026), and the only suicide with schizophrenia who died in the spring/summer was also the only one without positive symptomology. Our study is limited to male suicide completers, and results should not be generalized to women. We conclude that seasonal variation in suicide manifests itself differently in patients with different psychopathology. These findings indicate that assessment of suicide risk may need to include consideration of possible seasonal effects, depending on psychopathology. PMID- 15332196 TI - Alexithymia and suicidal ideation: a 12-month follow-up study in a general population. AB - Alexithymic features are often associated with depression, which is the most important risk factor for suicidal behaviors. Nevertheless, little is known about the associations between alexithymia and suicidality. In this 12-month follow-up study we investigated the relationship between alexithymia and suicidal ideation in a sample of the general population (N = 1,563) using the 20-item Toronto Alexithymia Scale (TAS-20) and the 21-item Beck Depression Inventory (BDI). Suicidal ideation was more common among subjects with alexithymia than among nonalexithymic subjects (32% v 9% at baseline and 36% v 9% after 12 months). In cross-sectional analyses, alexithymia associated with the presence of suicidal ideation even after adjustment for sex, age, and several psychosocial and socioeconomic factors and the presence of depression. Moreover, after adjustment for depression at baseline, the decrease and increase in alexithymic features during the study period associated independently with recovery from and the occurrence of suicidal ideation, respectively. Nevertheless, these associations were no longer independent when adjusted for concomitant changes in the level of depressive symptoms. In conclusion, if depression presents alexithymic features the subject has an additive impact on the risk of suicidal ideation. PMID- 15332197 TI - Test-retest and case-control study of psychological symptoms and social adjustment following parasuicide. AB - Numerous studies have been conducted in which cases of parasuicide have been followed for repeat parasuicide. By comparison, there have been relatively few studies of other morbidity endpoints. Over a 1-year period, 507 cases of parasuicide and 200 controls were recruited from emergency departments in Edmonton, Canada and interviewed using the Beck Hopelessness Scale, Beck Depression Inventory, State Anger Scale, Trait Anger Scale, Self-Esteem Scale, and Social Adjustment Scale. Over the ensuing 1 to 2 years, 414 cases were reinterviewed using the same questionnaires. Mean symptom scores on all questionnaires improved over the course of follow-up, but did not reach control levels. The pattern of improvement varied according to questionnaire. Only half of cases received treatment for the parasuicide from a psychiatrist, psychologist, or other health care provider during the 3 months following the emergency department visit. PMID- 15332198 TI - Personality and attempted suicide in depressed adults 50 years of age and older: a facet level analysis. AB - We examined the contribution of personality traits to attempted suicide, the number of suicidal attempts, and suicidal ideation in a sample of depressed inpatients. Personality was assessed via the Revised NEO Personality Inventory (NEO-PI-R). Bivariate analyses showed that suicide attempters were more self conscious, self-effacing, impulsive, and vulnerable to stress, and less warm, gregarious, and inclined to experience positive emotions. Multivariate regression analyses controlling for age, gender, severity of depression, and psychiatric comorbidity showed that patients with a lifetime history of attempted suicide were less inclined to experience positive emotions and be more self-effacing. Patients with more severe suicidal ideation were less warm and more self effacing. Results indicated that specific personality traits confer risk for suicidal behaviors in middle age and older adults. Interventions tailored to specific personality profiles in this high-risk group should be developed, and their efficacy examined. PMID- 15332199 TI - Gender-related differences among Turkish patients with obsessive-compulsive disorder. AB - The purpose of the present study was to investigate gender-related sociodemographic and clinical differences among Turkish patients with obsessive compulsive disorder (OCD). A total of 169 patients diagnosed with OCD by DSM-III R or DSM-IV criteria were included in this study. Male (n = 73) and female (n = 96) OCD patients were compared with respect to the demographic variables and the scores obtained from the Hamilton Rating Scale for Anxiety (HRSA), the Hamilton Rating Scale for Depression (HRSD) and the Yale-Brown Obsessive Compulsive Scale (Y-BOCS). We found a significantly earlier age at onset in male patients. No significant difference in terms of HARS, HDRS, and Y-BOCS scores was detected between the two groups. We observed a significantly higher frequency of contamination obsessions in females, and that of aggression and sexual obsessions in males. There was no significant difference in terms of the frequency of compulsions between the two groups. We also found that compulsion severity on obsessions/compulsions was higher in females and comorbidity rates of social phobia and schizophrenia were higher in males. Considering our results in combination with those of other studies, similarities rather than differences in gender-related sociodemographic and clinical characteristics of OCD patients across different populations seem to be present. PMID- 15332200 TI - Twelve-month prevalence of obsessive-compulsive disorder in Konya, Turkey. AB - We conducted a household survey of 3,012 adults aged 18 and over in order to estimate the prevalence of DSM-IV obsessive-compulsive disorder (OCD) in urban areas in Konya, Turkey. Trained psychiatry interns administered the 2.1 version of the OCD section of the Composite International Diagnostic Interview (CIDI). The 12-month period prevalence rate of OCD was 3.0%. The mean age of onset of OCD was 25.9 +/- 12.5 (range, 7 to 63) years. The prevalence rate of OCD was slightly higher among females (males 2.5%, females 3.3%), but the difference was not statistically significant. The relative risk for divorced, separated, or widowed subjects was approximately 4.2 times higher for OCD than others (2.7% v 10.5%). The 1-year prevalence of OCD inversely related to age group in male subjects, but increased with age in female subjects. The prevalence rate of OCD was not different by the level of education, except it was statistically higher among subjects who were literate but had no schooling, of which the causal relationship was high prevalence rate of OCD among female literate-but no schooling subjects. Subjects with few (one or two) and more siblings (seven or more) had a significantly higher prevalence rate of OCD than subjects with moderate numbers of siblings (three to six). No significant difference was found according to employment, fertility, birth order, and income of the subjects. About 30% of subjects with OCD had only obsessions, whereas 68.5% had both obsessions and compulsions. Only one subject (1.1%) with OCD met compulsion criteria without obsessions. The prevalence rate of OCD we found in Konya, Turkey was similar to the prevalence rates of most epidemiological studies. PMID- 15332201 TI - Patients with obsessive-compulsive disorder and hoarding symptoms: a distinctive clinical subtype? AB - We investigated whether patients with obsessive-compulsive disorder (OCD) and hoarding symptoms can be differentiated from their counterparts with other types of obsessions and compulsions in terms of sociodemographic and clinical features. Ninety-seven patients with OCD were assessed with a sociodemographic and clinical questionnaire, the Structured Clinical Interview for DSM-IV (SCID-I), the Yale Brown Obsessive-Compulsive Scale (Y-BOCS), the Beck Depression Inventory (BDI), the Hamilton Rating Scale for Depression (HDRS), and the Global Assessment of Functioning (GAF). Fifteen patients who reported hoarding symptoms in the Y-BOCS checklist (15.6% of the total sample) were compared and contrasted with 82 patients without those symptoms using the Mann-Whitney U test for continuous variables and the Pearson's goodness-of-fit chi-square test for categorical ones; Fisher's exact test was employed when indicated. Hoarders were characterized by (1) higher educational levels (chi(2) = 7.49; df = 2; P =.02); (2) earlier age at onset (Z = -2.99; P =.003); (3) higher rates of symmetry obsessions (chi(2) = 7.03; df = 1; P =.01); (4) greater frequency of ordering (chi(2) = 10.08; df = 1; P =.004); (5) rituals repetition (chi(2) = 4.42; df = 1; P =.03); (6) counting compulsions (chi(2) = 5.92; df = 1; P =.02); and (7) significantly higher rates of comorbidity with bipolar II disorder (chi(2) = 10.62; df = 1; P =.02) and (8) with eating disorders (chi(2) = 7.42; df = 1; P =.02). In conclusion, patients with OCD exhibiting hoarding feature a distinctive sociodemographic and clinical profile. It remains to be investigated whether these phenotypical characteristics are underlined by specific neurobiological mechanisms. PMID- 15332202 TI - Dissociative experiences in obsessive-compulsive disorder and trichotillomania: clinical and genetic findings. AB - A link between dissociation proneness in adulthood and self-reports of childhood traumatic events (including familial loss in childhood, sexual/physical abuse and neglect) has been documented. Several studies have also provided evidence for an association between dissociative experiences and trauma in patients with various psychiatric disorders, including post-traumatic stress disorder, borderline personality, dissociative identity and eating disorders. Based on the relative paucity of data on dissociation and trauma in obsessive-compulsive disorder (OCD) and trichotillomania (TTM), the primary objective of this study was to examine the relationship between trauma and dissociative experiences (DE) in these two diagnostic groups. Furthermore, the availability of clinical and genetic data on this sample allowed us to explore clinical and genetic factors relevant to this association. A total of 110 OCD and 32 TTM patients were compared with respect to the degree of dissociation (using the Dissociative Experiences Scale [DES]) and childhood trauma (using the Childhood Trauma Questionnaire [CTQ]). Patients were classified on the DES as either "high" (mean DES score >/= 30) or "low" (mean DES score < 30) dissociators. Additional clinical and genetic factors were also explored with chi-square and t tests as appropriate. A total of 15.8% of OCD patients and 18.8% of TTM patients were high dissociators. OCD and TTM groups were comparable on DES and CTQ total scores, and in both OCD and TTM groups, significant positive correlations were found between mean DES scores and mean CTQ subscores of emotional abuse, physical abuse, sexual abuse, and physical neglect. In the OCD group, high dissociators were significantly younger than low dissociators, and significantly more high dissociators than low dissociators reported a lifetime (current and past) history of tics (P <.001), Tourette's syndrome (P =.019), bulimia nervosa (P =.003), and borderline personality disorder (P =.027). In the TTM group, significantly more high dissociators than low dissociators reported (lifetime) kleptomania (P =.005) and depersonalisation disorder (P =.005). In the Caucasian OCD patients (n = 114), investigation of genetic polymorphisms involved in monoamine function revealed no significant differences between high and low dissociator groups. This study demonstrates a link between childhood trauma and DE in patients with OCD and TTM. High dissociative symptomatology may be present in a substantial proportion of patients diagnosed with these disorders. High dissociators may also be differentiated from low dissociators on some demographic features (e.g., lower age) and comorbidity profile (e.g., increased incidence of impulse dyscontrol disorders). Additional work is necessary before conclusions about the role of monoaminergic systems in mediating such dissociation can be drawn. PMID- 15332203 TI - Relationship between psychopathology and cognitive functioning in schizophrenia. AB - The purpose of this study was to delineate the relationship between positive, negative, cognitive, depressive, and excitement symptom dimensions of schizophrenia and cognitive functioning. Fifty-eight patients with schizophrenia (DSM-IV criteria) were assessed using the Positive and Negative Syndrome Scale (PANSS) and a battery of neuropsychological tests (executive function/abstraction, verbal and spatial working memory, verbal and nonverbal memory/learning, attention, visuospatial ability, and psychomotor speed). The cognitive symptom dimension correlated with executive functions, attention, verbal memory, and spatial ability. Severity of the negative symptom dimension was related to impairment in the structure of the semantic knowledge system, verbal memory, and auditory attention. In contrast, severity of the positive symptom dimension correlated only with impairment in the structure of the semantic knowledge system, and psychomotor speed. Finally, severity of the depressive and excitement symptom dimension was not associated with cognition. Correlations between symptom dimensions and cognitive measures were at best modest. Severity of cognitive and negative symptoms was mainly correlated with deficits on executive functions, semantic memory, and verbal memory, while positive symptoms only with semantic memory. These correlations were modest, suggesting that psychopathology and cognitive deficits in schizophrenia are caused, at least partially, by distinct pathophysiological processes. PMID- 15332204 TI - Stress in adult students with schizophrenia in a supported education program. AB - The successful integration of former psychiatric inpatients into the community requires innovative programs of psychosocial rehabilitation, including supported education. This article examines psychological distress as an outcome variable, and social support and coping strategies as mediating variables among 70 service user students (SUS) with schizophrenia and a comparison group of 55 adult students (AS) with no psychiatric diagnosis. Both groups were participants in a supported education program. The study variables were assessed by standardized research instruments: the Talbieh Brief Distress Inventory (TBDI), the Multidimensional Scale of Perceived Social Support (MSPSS), and the Coping Inventory for Stressful Situations (CISS). Univariate and multivariate analyses were used. Compared with the control subjects, SUS reported higher emotional distress and the utilization of emotion-oriented coping strategies, and a lesser availability of social support from family and friends. These variables explained 46.3%, 24.5%, and 22.5%, respectively, of the total variance in psychological distress scores. The findings provide the basis for interventions geared to reduce distress and, as a result, to enable students with severe mental illness to fully utilize the supported education program. PMID- 15332205 TI - Patterns of loneliness in an immigrant population. AB - Loneliness has been recognized as a public health problem that requires the attention of clinicians and researchers both as a condition in itself and in its relation to other conditions. This study sought to examine the relationship between self-reported loneliness, psychological distress, and social support among immigrants. A community survey of 386 recent immigrants to Israel from the former Soviet Union was conducted using the Revised UCLA Loneliness Scale (R-UCLA LS), Talbieh Brief Distress Inventory (TBDI), and Multidimensional Scale of Perceived Social Support (MSPSS). A cross-sectional design, and correlation and factor analyses were used to study the relationship between the studied variables. The distress-related and distress-free patterns of loneliness were distinguished as independent constructs, each with a specific sphere of influence. Distress-related loneliness accounted for 56.3% and distress-free for 18.2% of the total variance in individual loneliness scores. Distress-related loneliness is a generalized negative experience embedded in an array of distress symptoms, while distress-free loneliness appears to be a normal psychological reaction to dissatisfaction with current friend support. An important implication of this study in mental health practice is the sensitivity to these differences when treating recent immigrants. PMID- 15332206 TI - Demographic, clinical, and neurocognitive predictors of quality of life in schizophrenia patients receiving conventional neuroleptics. AB - We sought to evaluate demographic, clinical, and neurocognitive predictors of self-rated life quality and hospitalization in schizophrenia patients without the potentially cognition-enhancing influence of newer generation neuroleptic medication. A sample of 55 atypical neuroleptic-naive schizophrenia patients was assessed at index and 3 years later. Index neurocognitive measures included general intellectual ability (IQ), executive ability (Wisconsin Card Sorting Test [WCST]), verbal memory (California Verbal Learning Test [CVLT]), and manual dexterity (Purdue Pegboard). These measures, along with demographic (age, sex, education) and clinical (symptoms, prior hospitalizations) variables, were entered into regression equations to predict life quality (Sickness Impact Profile [SIP]) at follow-up, as well as rehospitalization during the 3-year period. Stability data were also analyzed. Demographic and cognitive data predicted subjective quality of life, but not rehospitalization. Changes in memory over time rather than performance levels related to life quality at follow up. Rehospitalization was related only to demographic data and previous hospital admissions. The findings support the predictive value of selected aspects of neurocognition in relation to a subjective outcome domain in schizophrenia. PMID- 15332211 TI - Immunosuppressive treatment for idiopathic membranous nephropathy: a systematic review. AB - BACKGROUND: This study aims to assess whether immunosuppression is beneficial in the treatment of idiopathic membranous nephropathy (IMN). METHODS: We reviewed randomized controlled trials (RCTs) addressing the effect of immunosuppression on histologically proven IMN in adults with nephrotic syndrome followed up for at least 6 months. The literature was searched according to Cochrane Collaboration guidelines. Four therapeutic classes were considered: (1) steroids (alone), (2) alkylating agents (alone or in combination with steroids), (3) calcineurin inhibitors (alone or in combination with steroids), and (4) antiproliferative agents (alone). RESULTS: Eighteen RCTs were selected (1,025 patients). Overall, no differences were found between immunosuppressive treatment and placebo or no treatment. For oral glucocorticoid therapy, no beneficial effect was observed. For alkylating agents, a beneficial effect was observed on complete remission (relative risk [RR], 2.37; 95% confidence interval [CI], 1.32 to 4.25; P = 0.004). Excluding patients with particularly well-preserved renal function, there was no significant difference in complete remission. Cyclophosphamide therapy resulted in a lower rate of adverse-event discontinuations compared with chlorambucil (8 versus 21 discontinuations, respectively; RR, 2.34; 95% CI, 1.25 to 4.39; P = 0.008). For calcineurin inhibitors, no difference was observed. For antiproliferative agents, a paucity of data did not allow a conclusion. CONCLUSION: The meta-analysis failed to show a long-term effect of treatment on patient and/or renal survival. There is weak evidence that the immunosuppressive regimen increased the remission rate. The review of safety showed a higher number of discontinuations for adverse events in immunosuppressive-treatment groups and that cyclophosphamide had fewer side effects than chlorambucil. PMID- 15332213 TI - Prognosis of ARF in hospitalized elderly patients. AB - BACKGROUND: Differentiation between hospital-acquired acute renal failure (ARF) and community-acquired ARF may have epidemiological implications that lead to different prognoses in hospitalized patients. Such a comparison has not yet been made among elderly individuals. METHODS: We performed a 3-year prospective study in a tertiary referral hospital of 325 patients aged 60 years or older who presented with ARF. Patients were divided into 2 groups; those with hospital acquired ARF (n = 154) and community-acquired ARF (n = 171), and were followed up in relation to mortality. Multiple logistic regression was used in the analysis. RESULTS: The overall mortality rate in this elderly population was 54%; 59% for the group with hospital-acquired ARF and 41% for the group with community acquired ARF (P < 0.001). Groups differed (P < 0.01) in relation to cause of ARF, preexisting diseases, organ failure, sepsis, and performance of dialysis, among other factors. The adjusted mortality risk for the group with hospital-acquired ARF was 2.23 times greater than for the group with community-acquired ARF (95% confidence interval [CI], 1.21 to 4.08). In the group with hospital-acquired ARF, factors associated with mortality were neurological failure (odds ratio [OR], 2.97; 95% CI, 1.17 to 7.60), hematologic failure (OR, 4.30; 95% CI, 1.63 to 11.34), and oliguria (OR, 12.14; 95% CI, 4.62 to 31.87). In the group with community-acquired ARF, significant factors were neoplasia, cardiac disease, hepatic disease, cardiovascular failure, oliguria, and sepsis. CONCLUSION: Differentiation between hospital-acquired ARF and community-acquired ARF is important in determining the prognosis of ARF in the elderly. Mortality risk factors are different in these 2 groups, and knowledge of their characteristics may allow better management of such patients. PMID- 15332212 TI - High-dose furosemide for established ARF: a prospective, randomized, double blind, placebo-controlled, multicenter trial. AB - BACKGROUND: The effect of furosemide on the survival and renal recovery of patients presenting with acute renal failure (ARF) is still debated. METHODS: Three hundred thirty-eight patients with ARF requiring dialysis therapy were randomly assigned to the administration of either furosemide (25 mg/kg/d intravenously or 35 mg/kg/d orally) or matched placebo, with stratification according to severity at presentation. The primary end point was survival. The secondary end point was number of dialysis sessions. Tertiary end points included time on dialysis therapy, time to achieve a serum creatinine level less than 2.26 mg/dL (<200 micromol/L), and time to reach a 2-L/d diuresis. RESULTS: There were no differences in survival and renal recovery rates between the 2 groups. Time to achieve a 2-L/d diuresis was shorter with furosemide (5.7 +/- 5.8 days) than placebo (7.8 +/- 6.8 days; P = 0.004). Overall, 148 patients achieved a urine output of at least 2 L/d during the study period (94 of 166 patients; 57%) with furosemide versus 54 of 164 patients (33%) with placebo ( P < 0.001). However, there were no significant differences in number of dialysis sessions and time on dialysis therapy between the furosemide and placebo groups, even in the subgroup of patients reaching a 2-L/d diuresis. CONCLUSION: High-dose furosemide helps maintain urinary output, but does not have an impact on the survival and renal recovery rate of patients with established ARF. PMID- 15332214 TI - Diagnostic and prognostic value of plasma brain natriuretic peptide in non dialysis-dependent CRF. AB - BACKGROUND: Brain natriuretic peptide (BNP) is useful for the evaluation of ventricular dysfunction in patients with various cardiac diseases. However, its diagnostic value has been considered to be limited in patients with chronic renal failure (CRF) because renal dysfunction itself may affect BNP levels. This study is designed to clarify the diagnostic and prognostic value of plasma BNP level in patients with CRF. METHODS: In 103 non-dialysis-dependent patients with CRF without heart failure and 60 hypertensive patients with normal renal function, echocardiographic examinations and BNP measurements were performed. RESULTS: Plasma BNP level was much greater in patients with CRF than in hypertensive controls; however, multiple regression analysis showed that left ventricular (LV) end-diastolic volume (EDV) index (LVEDVI) and the difference in mitral and pulmonary venous atrial wave duration (Ad-PVad), a marker of LV end-diastolic pressure, were independent determinants of plasma BNP levels in patients with CRF. The influence of LV overload (LVEDVI > or = 75 mL/m 2 and/or Ad-PVad < 0 milliseconds) on plasma BNP levels in subjects with CRF was independent of the severity of renal dysfunction. From Kaplan-Meier event-free curves (mean follow up, 13 months), the incidence of heart failure was much greater in patients with a plasma BNP level of 150 pg/mL or greater ( P < 0.001). By means of multivariate Cox regression analysis, high plasma BNP level was the strongest predictor for heart failure events (hazard ratio, 6.31; P < 0.001). CONCLUSION: These findings support plasma BNP level as a reliable marker of LV overload, even in nondialysis patients with CRF. Also, a high BNP level (> or =150 pg/mL) may have powerful predictive potential for heart failure in these patients. PMID- 15332215 TI - Simultaneous transjugular renal biopsy and hemodialysis catheter placement in patients with ARF. AB - BACKGROUND: Previous studies have evaluated transjugular renal biopsy in patients with contraindications to percutaneous renal biopsy or those undergoing simultaneous renal and hepatic biopsies. We sought to evaluate transjugular renal biopsy in patients with acute renal failure (ARF) or ARF in the presence of chronic renal insufficiency (CRI) who required venous catheter placement for hemodialysis (HD). METHODS: Ten consecutive patients (6 patients, ARF; 4 patients, ARF on CRI) at a single tertiary-care medical center, while undergoing placement of HD access through the internal jugular route, also underwent transjugular renal biopsy using the Quick-Core (Cook, Bloomington, IN) system to delineate the cause of ARF. Transjugular renal biopsy was performed because it was the opinion of the attending nephrologist that a histological diagnosis might alter management. RESULTS: Renal biopsy findings were diabetic nephropathy (3 patients), acute tubular necrosis (ATN; 2 patients), nephrosclerosis (2 patients), immunoglobulin A nephropathy (1 patient), lupus nephritis with focal crescents and ATN (1 patient), and pauci-immune necrotizing glomerulonephritis (1 patient). There were no major complications from the procedures. Among the 6 patients with ARF, management was directly affected in 3 patients (either initiation of appropriate immunosuppressive therapy or withholding of such therapy). In the remaining 3 patients with ARF and in patients with ARF on CRI, performing transjugular renal biopsy at the time of HD access placement obviated additional testing and/or unnecessary therapy. Four patients recovered renal function and HD therapy was discontinued, 2 patients died, and 1 patient was lost to follow-up. CONCLUSION: Simultaneous transjugular renal biopsy/HD catheter placement should be considered in patients with ARF requiring HD therapy for whom knowledge of the renal histological diagnosis may alter patient management. PMID- 15332216 TI - Hyperhomocysteinemia and the development of chronic kidney disease in a general population: the Hisayama study. AB - BACKGROUND: Hyperhomocysteinemia has been linked with various atherosclerotic diseases, but has not been evaluated sufficiently as a risk factor for the development of chronic kidney disease (CKD) in the general population. METHODS: To clarify this issue, we followed up 1,477 community-dwelling individuals without CKD, aged 40 years or older, for 5 years and examined the effects of baseline serum total homocysteine (tHcy) levels on the development of CKD. RESULTS: During follow-up, 88 subjects experienced CKD. Baseline tHcy levels were greater in men than women (1.35 versus 1.04 mg/L [10.0 versus 7.7 micromol/L]; P < 0.01). Age-adjusted 5-year incidences were 2.2% in the low tertile, 5.4% in the middle tertile, and 8.6% in the high tertile of tHcy levels for men and 3.3%, 6.0%, and 6.9% for women, respectively. The difference between the low and high tertiles was statistically significant for both sexes ( P < 0.05). In multivariate analysis, these relationships remained substantially unchanged, even after adjustment for other confounding factors, such as systolic blood pressure, antihypertensive medication, hemoglobin A 1c level, total cholesterol level, high density lipoprotein cholesterol level, habitual smoker status, regular alcohol intake, proteinuria, and baseline kidney function (odds ratio [OR] in the high tertile of tHcy levels, 2.09; 95% confidence interval [CI], 0.66 to 6.61 for men; OR, 2.86; 95% CI, 1.10 to 7.43 for women). Furthermore, baseline tHcy level showed a significantly inverse association with rate of change in kidney function during the 5 years after being adjusted for confounding factors, including baseline kidney function. CONCLUSION: Our findings suggest that elevated serum tHcy levels are a significant risk factor for the development of CKD in the general population. PMID- 15332217 TI - Lipids enhance alpha1-adrenoceptor pressor sensitivity in patients with chronic kidney disease. AB - BACKGROUND: Dyslipidemia in patients with chronic kidney disease (CKD) may contribute to hypertension (HT) by altering neurocirculatory control. To test this notion, we quantified the effects of acutely increasing levels of nonesterified fatty acids (NEFAs) and triglycerides on alpha1-adrenergic pressor sensitivity and baroreflex sensitivity (BRS) in subjects without diabetes with CKD. METHODS: Alpha1 pressor sensitivity was determined before and after increasing NEFA and triglyceride levels with a fat emulsion and heparin infusion in 8 subjects with stage 2 to 3 CKD (glomerular filtration rate, 56 +/- 6 mL/min by 125 I-iothalamate clearance). Seven subjects with HT and 8 normotensive control (CO) subjects with normal renal function and matched to patients with CKD also were studied. RESULTS: Fasting NEFA levels were greater in patients with CKD than in CO subjects (585 +/- 98 versus 321 +/- 32 micromol/L; P < 0.01), but not different from those in subjects with HT (501 +/- 68 micromol/L). Alpha1 pressor sensitivity, defined as the phenylephrine dose that increased mean blood pressure (BP) by 20 mm Hg, was greatest (lowest dose) in subjects with CKD (0.83 +/- 0.11 microg x kg(-1) x min(-1)), followed by those with HT (1.08 +/- 0.19 microg x kg( 1) x min(-1)) and CO subjects (1.34 +/- 0.19 microg x kg(-1) x min(-1); P < 0.05 versus CKD group). BRS, measured as the ratio of change in R-R interval to change in systolic BP during the phenylephrine infusion, was lowest, intermediate, and highest in the CKD, HT, and CO groups, respectively. Increasing NEFA and triglyceride levels significantly enhanced alpha1 pressor sensitivity in all 3 groups and reduced BRS in the CKD and CO groups. CONCLUSION: These observations suggest that the dyslipidemia prevalent in patients with stage 2 to 3 CKD may contribute to HT by enhancing alpha1 pressor sensitivity and impairing baroreflex function. PMID- 15332218 TI - Hyperhomocysteinemia in relation to plasma free amino acids, biomarkers of inflammation and mortality in patients with chronic kidney disease starting dialysis therapy. AB - BACKGROUND: Plasma levels of total homocysteine (tHcy) and free amino acids (AAs) are influenced by nutritional status in patients with chronic kidney disease (CKD), whereas the role of chronic inflammation is not clear. METHODS: In a cross sectional analysis, fasting levels of plasma tHcy, total cysteine (tCys), AA, serum albumin (Alb), and several inflammation markers, including C-reactive protein (CRP), were analyzed in a cohort of 250 patients with CKD starting renal replacement therapy. Patients were followed up during a 4-year period to assess overall mortality in relation to basal tHcy level. RESULTS: Ninety-three patients (37%) with signs of inflammation (CRP > or = 1 mg/dL) had significantly lower levels of tHcy, tCys, and serum Alb than 157 noninflamed patients. tHcy and tCys levels correlated positively with serum Alb levels and negatively with CRP levels (rho = -0.24; P < 0.0001; rho = -0.15; P < 0.05, respectively) and other inflammation markers. tHcy and tCys correlated significantly with levels of several AAs. The presence of both inflammation and malnutrition was associated with lower tHcy levels than when malnutrition was present without inflammation. Multivariate analysis showed that serum Alb, CRP, plasma folate, and vitamin B12 levels were independently associated with tHcy levels after adjustment for other variables. tHcy, but not tCys, level was significantly greater in survivors than nonsurvivors, and Kaplan-Meier analysis showed that greater tHcy level was associated with better survival. CONCLUSION: Plasma tHcy and tCys levels are interrelated to plasma AA levels and were lower in patients with inflammation. Thus, inflammation may contribute to the reverse association between tHcy level and mortality in patients with CKD starting renal replacement therapy. PMID- 15332219 TI - Increased cardiovascular risk in long-term hemodialysis patients carrying deletion allele of ACE gene polymorphism. AB - BACKGROUND: Controversy continues about the relation of insertion (I)/deletion (D) polymorphism of the angiotensin-converting enzyme (ACE) gene to the genetic predisposition to develop such cardiovascular diseases as myocardial infarction, stroke, and other arteriosclerotic disorders. METHODS: To examine the relation of ACE gene polymorphism to risk for developing cardiovascular diseases in long-term hemodialysis patients, we followed up 534 patients on long-term hemodialysis therapy for 3 years after determining ACE genotype. RESULTS: Numbers of patients with the II, ID, and DD genotypes were 208, 245, and 81, and frequencies of the I and D alleles were 0.62 and 0.38, respectively. Background characteristics, such as age, sex, causative diseases of renal failure, and preexistence of cardiovascular diseases at the time of study entry, were similar among the 3 genotype groups. Serum ACE activity was significantly greater in the DD and ID groups than the II group; however, plasma angiotensin II concentrations were not significantly different among the 3 groups. During the 3 years of follow-up, 46 fatal and 167 nonfatal cardiovascular events occurred. The incidence of these cardiovascular events was significantly associated with older age (P < 0.001), diabetes (P < 0.001), preexistence of cardiovascular diseases (P < 0.001), systolic blood pressure (P = 0.009), high cardiothoracic ratio on chest roentgenogram (P < 0.001), electrocardiographic abnormalities (P < 0.001), and low serum sodium level (P < 0.001). In addition, the incidence of cardiovascular events was greater in patients carrying the D allele (II, 22.1%; ID, 31.8%; DD, 38.3%; P = 0.010). CONCLUSION: It is suggested that the D allele of ACE gene polymorphism is a risk factor for cardiovascular complications in hemodialysis patients. PMID- 15332220 TI - Predicting the effect of intravenous calcitriol on parathyroid gland activity using double-phase technetium Tc 99m-sestamibi scintigraphy. AB - BACKGROUND: Although intravenous calcitriol is useful for decreasing intact parathyroid hormone (iPTH) blood levels in patients with secondary hyperparathyroidism (SHP) undergoing hemodialysis, approximately half these patients remain refractory to this treatment. The current study measures the diagnostic utility of double-phase technetium Tc 99m-sestamibi (MIBI) scintigraphy in predicting the response to calcitriol treatment. METHODS: Sixty hemodialysis patients with SHP with iPTH blood levels between 240 and 600 pg/mL (ng/L) were selected. Initial intravenous calcitriol pulse therapy was 6 microg/wk (for iPTH levels of 400 to 600 pg/mL [ng/L]) or 3 microg/wk (for iPTH levels of 240 to 400 pg/mL [ng/L]). MIBI scintigraphy was performed before the onset of calcitriol therapy and repeated 1 year later. Patients were injected intravenously with 740 MBq of MIBI. Images were obtained at 15 minutes (thyroid phase) and 2 hours (parathyroid phase) after radiotracer administration. Focal areas of increased MIBI uptake were considered pathological parathyroid glands. RESULTS: Forty-eight patients completed the study. After 1 year, iPTH levels had decreased significantly in 95.2% (47 patients), whereas good control (iPTH < 240 pg/mL [ng/L]) was reached in 70.8% (34 patients) and only 4 patients had iPTH levels greater than 400 pg/mL (ng/L; all were patients with 3 MIBI-positive areas at baseline determination). At baseline, there were 30 patients (62.5%) with MIBI positive areas (1, 2, or 3 areas), which decreased to 14 patients (29%) at the end of the study period. No patient showed 4 positive areas at any time. The 18 patients (37.5%) with no MIBI-positive area at baseline remained unchanged. CONCLUSION: MIBI scintigraphy is a reliable exploratory tool in predicting the response to treatment with intravenous calcitriol in hemodialysis patients with SHP. PMID- 15332221 TI - Total parathyroidectomy reduces elevated circulating fibroblast growth factor 23 in advanced secondary hyperparathyroidism. AB - BACKGROUND: Secondary hyperparathyroidism is a common complication in patients with stage 5 chronic kidney disease (CKD), accelerated by hyperphosphatemia. Fibroblast growth factor 23 (FGF-23), a phosphorus-regulating protein, has key roles in several phosphate-wasting disorders. The aim of this study is to examine the association of advanced secondary hyperparathyroidism with circulating FGF-23 levels. METHODS: Fifteen patients with marked secondary hyperparathyroidism (parathyroid hormone [PTH], 990 +/- 118 pg/mL [ng/L]) were enrolled. All underwent parathyroidectomy with forearm autotransplantation (PTX), and their FGF 23 levels were measured before and after PTX (days 1, 3, 7, and 10) by means of sandwich enzyme-linked immunosorbent assay. RESULTS: Preoperative FGF-23 levels correlated positively with phosphorus (P < 0.05), calcium-phosphorus product (Ca x P; P < 0.0005), and PTH values (P < 0.05). Serum FGF-23 levels decreased time dependently after PTX (P < 0.0005). Both serum phosphorus and Ca x P values decreased similarly after PTX ( P = 0.0001). Furthermore, FGF-23 levels days 1 and 3 correlated linearly with serum phosphorus (P < 0.05; P < 0.005, respectively) and Ca x P values (P < 0.01; P < 0.0001, respectively). CONCLUSION: FGF-23 levels correlate positively with serum phosphorus, Ca x P, and PTH values in patients with advanced secondary hyperparathyroidism. Complete ablation of progressive parathyroid glands reduces circulating FGF-23 levels, simultaneously decreasing serum phosphorus and Ca x P values. These findings suggest that hyperplastic parathyroid glands, together with hyperphosphatemia, affect abnormal FGF-23 metabolism in patients with stage 5 CKD with advanced secondary hyperparathyroidism. PMID- 15332222 TI - Religiosity in a hemodialysis population and its relationship to satisfaction with medical care, satisfaction with life, and adherence. AB - BACKGROUND: The religious beliefs and spirituality of patients on hemodialysis (HD) therapy have not been studied extensively. Studies of the dialysis population seem to indicate that religion may be associated with increased patient satisfaction with life and increased levels of social support. METHODS: Using multiple religiosity scales and scales to assess patient satisfaction with life and social support, we studied the relationship between religiosity and medical and/or social factors and adherence to treatment in 74 HD patients. RESULTS: High scores on the Intrinsic Religiosity Scale were associated strongly with high scores on the Satisfaction With Life Scale, whereas age and high Organizational Religious Activity Scale scores were associated strongly with high scores on the Satisfaction With Medical Care Scale. Older age was associated strongly with increased adherence. No relationship existed between religiosity and adherence in our population. CONCLUSION: Religious beliefs are related strongly to measures of satisfaction with life, whereas religious behaviors are related to satisfaction with medical care. Age is the single most important demographic factor associated with adherence. Because of the complex nature of religiosity, additional investigation is in order. PMID- 15332223 TI - Effect of differing blood lines on delivered blood flow during hemodialysis. AB - BACKGROUND The influence of differing blood tubing sets on delivered dialyzer blood flow (Q B ) was studied using ReadySet (RS; Medisystem Inc, Seattle, WA) and CombiSet (CS; Fresenius, Walnut Creek, CA). METHODS: In vitro, we measured Q B and pre-pump arterial pressures (P A; in millimeters of mercury) during 4 hours at a prescribed blood pump flow (Q P ) of 450 mL/min, using a glycerin-water mixture for RS and CS blood tubing sets (n = 10 each). We then performed 3 clinical studies to determine the difference between Q P and Q B as a function of P A for both tubing types and any effects of tubing type on delivered Kt/V in patients. RESULTS: In vitro, 4-hour time-averaged Q B was 451 mL/min with RS versus 422 mL/min with CS. Q B was approximately 40 mL/min greater using RS than CS blood tubing sets at Q P of approximately 445 mL/min with [Q P - Q B ] increasing linearly with P A for CS, but not RS (study 1). At a mean Q P of approximately 408 mL/min, CS blood tubing significantly increased Kt/V by approximately 0.1 (90 patients; study 2). In study 3, [Q P - Q B ] increased with treatment time with CS (8 to 29 mL/min), but not RS (from -4 to 6 mL/min). For patients achieving equal Q P of 400 mL/min or greater for all treatments (n = 11), paired analysis showed greater values for urea reduction ratio (+3.5%; P = 0.039) and Kt/V (+0.14; P = 0.04) with RS than CS tubing sets. CONCLUSION: Differences in [Q P - Q B ] occur both in vitro and in vivo, are much smaller with RS than CS tubing sets, and result in a greater delivered Kt/V of up to 0.14 for RS. PMID- 15332224 TI - Effects of different dialysis membranes on serum concentrations of epoetin alfa, darbepoetin alfa, enoxaparin, and iron sucrose during dialysis. AB - BACKGROUND: Numerous substances are administered during dialysis for anticoagulation and the treatment of anemia and iron deficiency. The availability of current dialysis membranes is diverse, but assessment of the effects of different membranes on these routinely administered substances is lacking. This prospective study appraises specific effects of 7 dialysis membranes (4 low-flux membranes, 3 high-flux membranes) on the handling of 4 commonly administered agents (epoetin alfa, darbepoetin alfa, enoxaparin, and iron sucrose) during dialysis. METHODS: Using a different membrane sequentially, 6 stable hemodialysis patients were treated with epoetin alfa, enoxaparin, and iron sucrose, and 6 patients with darbepoetin alfa. Serum concentrations (predialyzer and postdialyzer) of each substance were assessed after administration and after 4 hours of dialysis (predialyzer). RESULTS: Overall, use of low-flux membranes resulted in similar serum concentrations for all compounds. However, use of high flux membranes showed reduced antifactor Xa levels immediately (predialyzer, P = 0.028; postdialyzer, P = 0.027) and 4 hours after (P = 0.001) administration of enoxaparin compared with low-flux membranes. Variable changes in darbepoetin concentrations also were found between the high-flux membranes during dialysis (P = 0.009). Although peak serum concentrations of all compounds were inversely proportional to body weight, the percentage of change during dialysis was not related to dosage or body weight. CONCLUSION: High-flux membranes may require greater doses of enoxaparin to ensure adequate anticoagulation during dialysis and may be associated with variable changes in darbepoetin concentrations. For other compounds, no noteworthy difference among membranes was defined. PMID- 15332225 TI - Trends and dynamics of changes in calcification score over the 1-year observation period in patients on peritoneal dialysis. AB - BACKGROUND: Accelerated vascular calcification is an important cause of excess mortality in patients on dialysis therapy. The aim of the study was to evaluate the trends in coronary artery calcification (CAC) score (CaSc) during a 1-year period in a group of stable peritoneal dialysis (PD) patients and identify factors that may be associated with CaSc changes. METHODS: Sixty-one stable patients (28 women, 33 men) on PD therapy with a mean age of 50.4 +/- 13.6 years were included. Forty-seven patients survived the entire study period on PD therapy and were suitable for the final analysis. CaSc was assessed at baseline and after 12 months by using multislice spiral computed tomography. Proinflammatory cytokines (interleukin-6, tumor necrosis factor-alpha [TNF alpha]), acute-phase proteins (C-reactive protein [CRP], fibrinogen), calcium phosphate balance, and lipid profile were assessed at baseline and after 6 and 12 months. RESULTS: Median CaSc was 22.6 Agatston units (range, 0 to 5,502.8 Agatston units) at baseline and increased to 84 Agatston units (range, 0 to 5,001.3 Agatston units) at a 1-year follow-up (P < 0.05). In the entire group of patients, 3 subgroups were identified: patients with progression (n = 21; P = 0.02 for the difference between initial versus follow-up CaSc), patients with regression (n = 12; P = 0.05), and subjects without change in CaSc after 1 year (n = 14). Patients without progression showed no calcifications at baseline and follow-up and were younger, less overweight, and characterized by significantly lower mean TNF-alpha, leptin, and CRP levels during 1 year compared with both progressors and regressors. Mean serum phosphate and calcium x phosphate product (Ca x P) values were gradually increasing from regressors through the no calcification group to progressors (P < 0.01 for phosphate levels, P < 0.02 for Ca x P product). Significant correlations were found between changes in CaScs and mean values for phosphate (R = 0.44; P < 0.0005) and Ca x P product (R = 0.38; P < 0.005). CONCLUSION: Chronic nonspecific inflammation does not directly attribute to progression in CaScs. Calcium-phosphate balance abnormalities appear to be the only important factors promoting CAC, although a permissive or promoting role of inflammation cannot be ruled out. PMID- 15332226 TI - Transplantation in the diabetic patient with advanced chronic kidney disease: a task force report. AB - On May 7 to 8, 2003, more than 60 physicians, surgeons, nurses, and allied health professionals met in Chicago, IL, to discuss state-of-the-art approaches to transplantation in the clinical management of patients with diabetes and advanced chronic kidney disease. Four work groups addressed pretransplantation evaluation and management, therapeutic alternatives for treatment of patients with chronic kidney disease, posttransplantation diabetes mellitus, and improving long-term kidney transplant outcomes in patients with diabetes. This report summarizes the deliberations and recommendations of the task force. PMID- 15332227 TI - Fenofibrate-associated reversible acute allograft dysfunction in 3 renal transplant recipients: biopsy evidence of tubular toxicity. AB - Renal transplant recipients are susceptible to hyperlipidemia and the development of atherosclerosis as a consequence of the immunosuppressive agents they require, which include corticosteroids, calcineurin inhibitors, and sirolimus. Fibric acid derivatives and 3-hydroxymethylglutaryl-coenzyme A reductase inhibitors are prescribed commonly to optimize lipid profiles and reduce the risk of cardiovascular events in this type of setting. The authors describe 3 cases of reversible acute renal allograft dysfunction in patients treated with fenofibrate. Serum levels of monitored immunosuppressant agents remained therapeutic throughout the time period. Discontinuation of the fenofibrate resulted in the resolution of renal dysfunction. The pathologic changes to the proximal tubules in all 3 biopsy specimens were in keeping with a toxic rather than an ischemic etiology. Although the control of hyperlipidemia is crucial in the transplant patient population, the authors suggest that caution be exercised and serum creatinine levels be closely monitored in patients started on fibrates. PMID- 15332228 TI - Reversal of dialysis-dependent renal failure in light-chain deposition disease by autologous peripheral blood stem cell transplantation. AB - Specific treatment of light-chain deposition disease has been reported as ineffective in altering the course of the severe or end-stage renal failure it causes. The authors describe a case of biopsy-proven primary light-chain deposition disease of the kidney, severe renal failure, and incipient dialysis dependency, treated by autologous peripheral blood stem cell transplantation, that led to reversal of dialysis dependency and sustained improvement in renal function. PMID- 15332229 TI - A patient without monocytes who had pulmonary renal syndrome. AB - Clinical disorders with an isolated lack of monocytes have not been reported hitherto. The authors describe the case of a 38-year-old woman with pulmonary alveolar proteinosis and nephrotic syndrome caused by membranous nephropathy and widespread papillomatosis of the vulva. Immunologic studies showed normal levels of immunoglobulins and C2, C3c, and C4. Cryoglobulins and paraproteins were not detected. Antinuclear antibodies, antineutrophil cytoplasmic antibodies, and antiglomerular basement membrane antibodies were not detectable. Circulating immune complexes containing C1q, immunoglobulin G, and immunoglobulin M were elevated. The patient showed immunodeficiency that was characterized by complete anergy to intracutaneously administered recall antigens in vivo and to recall antigens in vitro. The immunodeficiency was accompanied by the absence of monocytes in the peripheral blood as well as in bone marrow cultures. In parallel, long-term bone marrow cultures and colony-forming cell assays did not result in the growth of monocytes. Mitogenic agents that require the presence of monocytes induced almost no T-cell proliferation (Concanavalin A: 5,841 counts per minute [cpm]), whereas agents that act directly on T cells induced intense T cell proliferation (phytohemagglutinin: 110,001 cpm; OKT 3: 120,616 cpm; and pokeweed mitogen: 89,474 cpm). These data suggest that the pulmonary renal syndrome in this patient results from the lack of monocytes and the consecutive defect of antigen presentation and antigen clearance. PMID- 15332230 TI - A reevaluation of empiric therapy for peritoneal dialysis-related peritonitis. PMID- 15332231 TI - The treatment of idiopathic membranous nephropathy: a dilemma or a conundrum? PMID- 15332234 TI - Opposing mechanisms in tubuloglomerular feedback. PMID- 15332232 TI - A woman with chronic hepatitis C infection and nephrotic syndrome who developed multiple renal lesions after interferon alfa therapy. PMID- 15332235 TI - Quiz page. Anti-GBM antibody-mediated glomerulonephritis with superimposed ANCA associated vasculitis. PMID- 15332237 TI - Crescentic glomerulonephritis and dense deposit disease in a woman with breast carcinoma on immunosuppressive chemotherapy. AB - The authors describe an unusual case of acute renal failure in a 50-year-old woman with a history of breast carcinoma. The breast carcinoma was treated with 4 cycles of chemotherapy. After chemotherapy, she felt fatigued and noticed decreased urine output. Her serum creatinine level had risen from 0.8 to 10.1 mg/dL (71 to 893 micromol/L). Renal biopsy was done that showed a severe crescentic glomerulonephritis secondary to dense deposit disease. This case is extremely unusual in that: (1) Dense deposit disease developed in a 50-year-old woman, whereas it is primarily a disease of children and young adults, and (2) the patient was being treated with immunosuppressive chemotherapy for breast carcinoma when dense deposit disease developed, thus posing a therapeutic dilemma. PMID- 15332238 TI - Hemodialysis-related portal-systemic encephalopathy. AB - The authors encountered a case of portal-systemic venous shunt newly diagnosed after initiation of hemodialysis. A 68-year-old Japanese woman began hemodialysis because of symptoms of uremia including loss of appetite and pulmonary edema. Loss of consciousness occurred suddenly after her ninth session of hemodialysis. No hepatic functional abnormality was found other than hyperammonemia (314 microg/dL [184 micromol/L]). Loss of consciousness subsequently occurred often after hemodialysis. Color Doppler ultrasonography and magnetic resonance angiography depicted a large shunt between the left gastric vein and left renal vein resulting in portal flow entering the systemic circulation via the renal vein. Because the shunt was large, ligation of it was performed surgically. Results of histologic examination of a liver biopsy specimen obtained intraoperatively were normal. The patient became well postoperatively. This patient's encephalopathy appeared to be caused by the flow of ammonia-rich portal venous blood into the systemic circulation via the large shunt owing to a decrease in intravenous pressure after rapid hemodialysis. Portal-systemic shunt encephalopathy should be recognized as a "new" neuropsychiatric disorder characteristic of patients undergoing hemodialysis. PMID- 15332239 TI - Squamous cell carcinoma in situ arising at the exit site of a tunneled catheter. AB - Squamous cell carcinoma (SCC) is a well-known complication of immunosuppression associated with organ transplantation. It may arise de novo or from previously existing in situ lesions (Bowen's disease). Concurrent human papilloma virus infection often has an etiologic role, and SCC may follow an aggressive course in immunosuppressed patients. The authors describe a liver transplant patient in whom end-stage renal disease developed. She underwent tunneled catheter placement followed by arteriovenous graft placement. Subsequently, a large SCC in situ at the exit site of her prior tunneled hemodialysis catheter occurred. The growth was removed surgically without complication. SCC has not been reported previously to arise from a catheter exit site. This entity is common in renal and other transplant populations and may follow an aggressive course. It should be sought out by careful skin evaluation to include areas not routinely exposed to the sun. PMID- 15332240 TI - Quantitative assessment of regional pulmonary perfusion in the entire lung using three-dimensional ultrafast dynamic contrast-enhanced magnetic resonance imaging: Preliminary experience in 40 subjects. AB - PURPOSE: To assess regional differences in quantitative pulmonary perfusion parameters, i.e., pulmonary blood flow (PBF), mean transit time (MTT), and pulmonary blood volume (PBV) in the entire lung on a pixel-by-pixel basis in normal volunteers and pulmonary hypertension patients. MATERIALS AND METHODS: Three-dimensional ultrafast dynamic contrast-enhanced MR imaging was performed in 15 normal volunteers and 25 patients with pulmonary hypertension. From the signal intensity-time course curves, PBF, MTT and PBV maps were generated using deconvolution analysis, indicator dilution theories, and the central volume principle, on a pixel-by-pixel basis. From pulmonary perfusion parameter maps of normal volunteers and pulmonary hypertension patients, regional PBF, MTT, and PBV were statistically evaluated. RESULTS: Regional PBF, MTT, and PBV showed significant differences in the gravitational and isogravitational directions (P < 0.05). The quantitative pulmonary perfusion parameter maps demonstrated significant differences between normal volunteers and pulmonary hypertension patients (P < 0.05). CONCLUSION: Three-dimensional ultrafast dynamic contrast enhanced MR imaging is feasible for the assessment of regional quantitative pulmonary perfusion parameters in the entire lung on a pixel-by-pixel basis in normal volunteers and pulmonary hypertension patients. PMID- 15332241 TI - Quantitative 1H-MRS of healthy human cortex, hippocampus, and thalamus: metabolite concentrations, quantification precision, and reproducibility. AB - PURPOSE: To evaluate metabolite concentrations in cortical gray matter, hippocampus, and thalamus of healthy adults, and to investigate precision and reproducibility of quantitative proton magnetic resonance spectroscopy (1H-MRS) in these gray matter regions. MATERIALS AND METHODS: Quantitative single-voxel short echo-time spectra were obtained from healthy human cortex, hippocampus, and thalamus. Subjects were examined twice. Metabolite concentrations, quantification precision, and reproducibility were determined. RESULTS: There were no significant differences between test and retest measurements. Regional differences were observed with respect to metabolite concentrations, quantification precision, and reproducibility. Quantification precision and reproducibility of N-acetylaspartate and N-acetyl aspartylglutamate (tNAA), creatine and phosphocreatine (tCr), choline-containing compounds (Cho), and myo inositol (myo-Ins), were better than those of glutamate (Glu) and glutamine (Gln). Generally, precision and reproducibility were better in cortex than in hippocampus or thalamus. The quantification precision was shown to correlate both with reproducibility and spectral linewidth. CONCLUSION: The reliability of quantitative MRS depends on the metabolite concerned, its concentration, and on the brain area studied. Moreover, the quantification precision of a metabolite in a single spectrum appears to be a reliable measure for its reproducibility in a longitudinal study. PMID- 15332242 TI - Reliability in detection of hemorrhage in acute stroke by a new three-dimensional gradient recalled echo susceptibility-weighted imaging technique compared to computed tomography: a retrospective study. AB - PURPOSE: To compare the sensitivity of magnetic resonance (MR) susceptibility weighted imaging (SWI) with conventional MR sequences and computed tomography (CT) in the detection of hemorrhage in an acute infarct. MATERIALS AND METHODS: A series of 84 patients suspected of having acute strokes had both CT and MR imaging (MRI) scans with diffusion-weighted imaging (DWI) and SWI. The SWI sequence is a new high-resolution three-dimensional (3D) imaging technique that amplifies phase to enhance the magnitude contrast. RESULTS: Thirty-eight of 84 cases showed abnormal DWI consistent with acute infarct. Of the 38, SWI showed evidence of hemorrhage in 16 cases, compared to eight cases with spin echo (SE) T2, seven cases with fluid attentuated inversion recovery (FLAIR), and only five cases with CT. In a subset of 17 cases of acute infarct who had both two dimensional gradient recalled echo (2D-GRE) T2*-weighted imaging and SWI, in addition to conventional MRI, evidence of hemorrhage was seen in 10 cases using SWI, compared to seven cases with 2D-GRE T2*. CONCLUSION: SWI proved to be a powerful new approach for visualizing hemorrhage in acute stroke compared to CT and conventional MRI methods. PMID- 15332243 TI - Three-dimensional magnetic resonance imaging technique for myocardial-delayed hyperenhancement: a comparison with the two-dimensional technique. AB - PURPOSE: To compare two-dimensional and three-dimensional techniques in the detection of myocardial infarction (MI) and in the grading transmural extent (TE). MATERIALS AND METHODS: Twelve patients with clinically proven MI were examined using two-dimensional and three-dimensional techniques with cardiac gated, breath-hold, T1-weighted gradient echo sequence with an inversion recovery pulse following gadopentetate dimeglumine (Gd-DTPA) at 0.2 mmol/kg. Contrast-to noise, signal-to-noise, and signal intensity ratios (CNR, SNR, and SIR, respectively) were derived and compared for each technique. RESULTS: From two dimensional to three-dimensional, statistical significant difference was found in the mean CNR (11.65 vs. 56.59; P = 0.002), SNR (18.03 vs. 76.90; P < 0.001), and SIR (3.6 vs. 6.36; P = 0.05). Intraobserver agreement (kappa) between two dimensional and three-dimensional were R1 = 74% and R2 = 90%. Interobserver agreements between the readers were two-dimensional = 77% and three-dimensional = 79%. CONCLUSION: Mean CNR, SNR, and SIR are significantly increased in the three dimensional technique compared to the conventional two-dimensional technique. PMID- 15332244 TI - Recurrent coronary artery stenosis: assessment with three-dimensional MR imaging. AB - PURPOSE: To assess the diagnostic value of three-dimensional coronary magnetic resonance (MR) angiography with fat saturation and navigator echo in the setting of restenosis after percutaneous transluminal coronary angioplasty (PTCA). MATERIALS AND METHODS: Thirty consecutive patients who had PTCA and were referred for elective coronary reangiography underwent MR imaging and coronary angiography. The pulse sequence was a cardiac triggered, single-slab, three dimensional, gradient-echo sequence, employing a spin-echo navigator echo measurement to track the variation of the diaphragm during the scan. The following segments of the coronary arteries were included in this prospective study: left main coronary artery, proximal and middle left anterior descending, proximal and middle left circumflex, proximal and middle right coronary artery, and intermediate branch, if present. The quality of the MR images was graded from 0 to 5. RESULTS: In total, 221 coronary artery segments could be identified. Mean image quality was 3.3. Overall accuracy for segments with an image quality of grade 2 or more was 90%. To achieve a positive predictive value >70% for a significant stenosis/restenosis, only segments with quality >/=3 could be assessed, whereas an acceptable negative predictive value could be achieved for nearly all segments. CONCLUSION: Our preliminary data suggest that MR coronary angiography may be most helpful as a screening test in selected patients to exclude clinically relevant stenoses or to assess restenoses after PTCA or in patients in whose coronary angiography is relatively contraindicated. PMID- 15332245 TI - Nitrate-induced coronary vasodilation by stress-magnetic resonance imaging: a novel noninvasive test of coronary vasomotion. AB - PURPOSE: To evaluate the feasibility of assessing coronary vasodilation following exogenous nitrates, using magnetic resonance angiography (MRA). The assessment of coronary response to exogenous nitrovasodilators may have a diagnostic and prognostic impact in patients with coronary artery disease. To date, stress imaging of coronary artery vasomotion has been confined to the catheterization laboratory. MRA is emerging as a noninvasive tool for coronary artery imaging. MATERIALS AND METHODS: Coronary MRA was performed in 20 healthy volunteers (12 males, age = 33 +/- 8). We used spiral spoiled gradient echo (SSGE) sequences for imaging of coronary artery lumen. After the baseline short-axis view of the coronary artery was obtained, sublingual nitroglycerin (NTG) (0.3 mg) was administered. In all subjects, short-axis views of the coronary artery were acquired repetitively (8-10 times) from 1 up to 10 minutes after NTG administration. Measurements were obtained by two independent investigators. RESULTS: Interpretable short-axis view of left anterior descending artery (LAD) was obtained in 15 subjects (75%); in the remaining five subjects the right coronary artery (RCA) was examined. The interobserver variability was 15%, and the intraobserver variability 4%. The NTG-induced maximal vasodilation was 43 +/- 22%. The vasodilator response over time after NTG was maximal on average at 279 +/- 112 seconds, but with substantial heterogeneity. CONCLUSION: Entity and time course of nitrate-induced coronary vasodilation in the left anterior descending and/or RCA can be assessed using MRA with high feasibility and reproducibility. Coronary MRA has potential for dynamic imaging of coronary vasomotion. PMID- 15332246 TI - Free-breathing, three-dimensional coronary artery magnetic resonance angiography: comparison of sequences. AB - PURPOSE: To compare six free-breathing, three-dimensional, magnetization-prepared coronary magnetic resonance angiography (MRA) sequences. MATERIALS AND METHODS: Six bright-blood sequences were evaluated: Cartesian segmented gradient echo (C SGE), radial SGE (R-SGE), spiral SGE (S-SGE), spiral gradient echo (S-GE), Cartesian steady-state free precession (C-SSFP), and radial SSFP (R-SSFP). The right coronary artery (RCA) was imaged in 10 healthy volunteers using all six sequences in randomized order. Images were evaluated by two observers with respect to signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), visible vessel length, vessel edge sharpness, and vessel diameter. RESULTS: C-SSFP depicted RCA over the longest distance with high vessel sharpness, good SNR, and excellent background suppression. S-GE provided best SNR and CNR in proximal segments, but more vessel blurring and poorer background suppression, resulting in poor visualization of distal segments. R-SSFP images showed good background suppression and best vessel sharpness, but only moderate SNR. C-SGE provided good SNR and reasonable CNR, but lowest vessel sharpness. S-SGE and R-SGE visualized the RCA over the smallest distance, mostly due to vessel blurring and low SNR, respectively. CONCLUSION: Overall, Cartesian SSFP provided the best image quality with excellent vessel sharpness, visualization of long vessel segments, and good SNR and CNR. PMID- 15332247 TI - Evaluation of heart perfusion in patients with acute myocardial infarction using dynamic contrast-enhanced magnetic resonance imaging. AB - PURPOSE: To investigate the diagnostic ability of quantitative magnetic resonance imaging (MRI) heart perfusion in acute heart patients, a fast, multislice dynamic contrast-enhanced MRI sequence was applied to patients with acute myocardial infarction. MATERIALS AND METHODS: Seven patients with acute transmural myocardial infarction were studied using a Turbo-fast low angle shot (FLASH) MRI sequence to monitor the first pass of an extravascular contrast agent (CA), gadolinium diethylene triamine pentaacetic acid (Gd-DTPA). Quantitation of perfusion, expressed as Ki (mL/100 g/minute), in five slices, each having 60 sectors, provided an estimation of the severity and extent of the perfusion deficiency. Reperfusion was assessed both by noninvasive criteria and by coronary angiography (CAG). RESULTS: The Ki maps clearly delineated the infarction in all patients. Thrombolytic treatment was clearly beneficial in one case, but had no effect in the two other cases. Over the time-course of the study, normal perfusion values were not reestablished following thrombolytic treatment in all cases investigated. CONCLUSION: This study shows that quantitative MRI perfusion values can be obtained from acutely ill patients following acute myocardial infarction. The technique provides information on both the volume and severity of affected myocardial tissue, enabling the power of treatment regimes to be assessed objectively, and this approach should aid individual patient stratification and prognosis. PMID- 15332248 TI - 3D breath-held cardiac function with projection reconstruction in steady state free precession validated using 2D cine MRI. AB - PURPOSE: To develop and validate a three-dimensional (3D) single breath-hold, projection reconstruction (PR), balanced steady state free precession (SSFP) method for cardiac function evaluation against a two-dimensional (2D) multislice Fourier (Cartesian) transform (FT) SSFP method. MATERIALS AND METHODS: The 3D PR SSFP sequence used projections in the x-y plane and partitions in z, providing 70 80 msec temporal resolution and 1.7 x 1.7 x 8-10 mm in a 24-heartbeat breath hold. A total of 10 volunteers were imaged with both methods, and the measurements of global cardiac function were compared. RESULTS: Mean signal-to noise ratios (SNRs) for the blood and myocardium were 114 and 42 (2D) and 59 and 21 (3D). Bland-Altman analysis comparing the 2D and 3D ejection fraction (EF), left ventricular end diastolic volume (LVEDV) and end systolic volume (LVESV), and end diastolic myocardial mass (LVEDM) provided values of bias +/-2 SD of 0.6% +/- 7.7 % for LVEF, 5.9 mL +/- 20 mL for LVEDV, -2.8 mL +/- 12 mL for LVESV, and 0.61 g +/- 13 g for LVEDM. 3D interobserver variability was greater than 2D for LVEDM and LVESV. CONCLUSION: In a single breath hold, the 3D PR method provides comparable information to the standard 2D FT method, which employs 10-12 breath holds. PMID- 15332249 TI - Use of a three-station phased array coil to improve peripheral contrast-enhanced magnetic resonance angiography. AB - PURPOSE: To explore the imaging capabilities of a new commercially available, three-station, 129-cm long, 12-element phased array coil for contrast-enhanced magnetic resonance angiography (CE-MRA) in patients with symptomatic peripheral arterial occlusive disease. MATERIALS AND METHODS: Nineteen patients, referred for peripheral CE-MRA, were evaluated using the new three-station coil. For each station four coil elements (two anterior and two posterior to the patient) were used. The expected improvements in signal-to-noise ratio (SNR) and contrast-to noise ratio (CNR) were used to improve spatial resolution and increase anatomic coverage for the distal two stations compared to our previous protocol. Images obtained in the 19 patients imaged with the new coil were compared to those of the last 19 patients scanned without the use of the new coil. Differences in image quality before vs. after the availability of the new coil were compared in terms of SNR and CNR, subjective interpretability score (SIS), degree of venous enhancement, and anatomic coverage. Images were interpreted by two experienced observers, blinded for imaging technique and each other's results. RESULTS: Use of the coil enabled acquisition of high resolution peripheral vasculature images in all cases and allowed for substantially smaller voxel sizes (thighs: 5.3 vs. 8.4 mm(3) [-37%]; legs: 1.8 vs. 8.0 mm3 [-78%]) and much shorter acquisition durations in the aortoiliac and thigh stations (aortoiliac: 16 vs. 27 seconds [ 41%]; thighs: 11 vs. 23 seconds [-52%]). Acquisition duration in the leg station was prolonged (68 vs. 29 seconds [+134%]). SNR and CNR were significantly higher only in the aortoiliac station using the three-station coil (both: P < 0.001). There were no significant differences in SIS for the aortoiliac and thigh stations (aortoiliac station: observer 1: P = 0.16, observer 2: P = 0.19; thigh station: both observers: P = 0.27). Images acquired with the new coil had significantly higher SIS for the leg station (both observers: P = 0.004). There were no significant differences in venous enhancement between the two protocols for any of the stations (all P > 0.11). In 12/12 (100%) requested cases the entire pedal arch was depicted using the new coil, whereas this was not possible with the old protocol. CONCLUSION: The new three-station dedicated peripheral vascular coil allows for much higher resolution imaging in the thigh and leg stations with greater anatomic coverage and substantially improves peripheral MRA quality of the lower leg vasculature. PMID- 15332250 TI - Angiogenesis in hepatocellular nodules: correlation of MR imaging and vascular endothelial growth factor. AB - PURPOSE: To assess the correlation between magnetic resonance (MR) imaging findings and angiogenetic activity in hepatocellular nodules evaluated by immunohistochemical staining with antibody of vascular endothelial growth factor (VEGF). MATERIALS AND METHODS: We searched the pathologic records of our institution from December 1999 to April 2002, and included 16 patients with hepatocellular carcinoma (N = 14), large regenerative nodule (N = 1), and dysplastic nodule (N = 1) who underwent orthotopic liver transplantation (10 patients) or partial hepatectomy (six patients) and MR imaging within an interval of two weeks. The MR images were retrospectively assessed qualitatively and quantitatively. Angiogenetic activity of the hepatic nodules was evaluated by means of immunohistochemical study for VEGF. Analysis of variance and the Scheffe criterion were used for statistical evaluation. RESULTS: Hepatic nodules with moderate to strong immunoreactivity for VEGF showed higher signal intensity on T1 weighted images (P < 0.05) and those with intense immunoreactivity for VEGF showed higher signal intensity on T2-weighted images (P < 0.05). No correlation was found between the immunoreactivity for VEGF and tumor vascularity on postcontrast early- and late-phase images. CONCLUSION: Our current results suggest that signal intensity on unenhanced T1- and T2-weighted MR images may correlate with immunoreactivity for VEGF. Correlation was not found between immunoreactivity for VEGF and signal intensity on gadolinium-enhanced MR images. PMID- 15332251 TI - Comparison of matched-filtered two-dimensional projection and elliptical centric ordered three-dimensional contrast-enhanced magnetic resonance angiography. AB - PURPOSE: To compare the image quality of matched-filtered two-dimensional projection magnetic resonance angiography (MRA) and elliptical centric-ordered (EC) three-dimensional MRA. MATERIALS AND METHODS: Signal-to-noise ratios (SNRs) of matched-filtered two-dimensional projection and EC three-dimensional MRA are developed theoretically and compared by clinical studies, in which 10-20 mL of gadolinium (Gd) was injected at 1.5 mL/second. The artery-vein contrast in two dimensional projection MRA was managed by manually selecting specific templates for the matched filters. RESULTS: The SNR of matched-filtered two-dimensional projection MRA is superior to that of EC three-dimensional MRA for vessels wider than one pixel due to the integral effect. The artery-vein contrast can be managed flexibly in two-dimensional projection MRA by choosing different templates for the matched filter, while the artery-vein contrast in EC three dimensional MRA is solely determined by the timing to start the acquisition. CONCLUSION: Matched-filtered two-dimensional projection MRA provides comparable image quality and is a flexible alternative to EC three-dimensional MRA in applications where contrast timing is difficult and temporal information is of interest. PMID- 15332252 TI - High-resolution MR-imaging of the liver with T2-weighted sequences using integrated parallel imaging: comparison of prospective motion correction and respiratory triggering. AB - PURPOSE: To compare high-resolution T2-weighted images of the liver with and without integrated parallel acquisition techniques (iPAT) using either breath hold sequences in combination with prospective acquisition motion correction (PACE) or respiratory triggering. MATERIALS AND METHODS: Ten volunteers and 10 patients underwent each four different high-resolution fast spin echo (FSE) T2 weighted sequences with 5 mm slice thickness and a full 320 matrix: a multi breath-hold FSE sequence with and without iPAT and PACE and a respiratory triggered FSE sequence with and without iPAT. Image quality was rated with a five point scale by two independent readers. Signal intensity measurements were performed on a water phantom. RESULTS: The sequences with iPAT required a substantially shorter acquisition time without loss of image quality. Overall image quality was rated equal for all sequences by both readers. Image time for nine slices with iPAT was 13 seconds (19 seconds without iPAT) with multi-breath hold and on average 4:00 minutes (7:02 minutes without iPAT) with respiratory triggering. Imaging with the PACE technique resulted in more correct positioning of the image stacks. CONCLUSION: T2-weighted fast imaging with iPAT is feasible and results in high-quality images within a short acquisition time. Overall image quality is not negatively affected by iPAT. PMID- 15332253 TI - Adnexal torsion: magnetic resonance findings in the viable adnexa with emphasis on stromal ovarian appearance. AB - PURPOSE: To evaluate ultrasound (US) and magnetic resonance (MR) findings in the viable twisted adnexa. MATERIALS AND METHODS: Ten patients underwent US and MR studies before surgical detorsion. Corrected cross-sectional area of the ovary was defined as cross-sectional area minus areas of cysts and follicles superior to 1 cm. On T2-weighted images, signal intensity of the stroma was graded as type 1 when it was equal to that of urine and type 2 when it was less than that of urine but markedly more than the contralateral side. RESULTS: The tube was twisted in six cases and the ovary in nine cases. All adnexa were viable. The largest ovarian cross-sectional area and the largest corrected ovarian cross sectional area of the twisted ovary were significantly larger than those of the contralateral ovary (P = 0.043 for US; P = 0.012 and 0.017, respectively, for MR). These ovaries contained types 1 and 2 hyperintensity in six cases and only type 2 hyperintensity in three cases. Tubal thickening was seen on MR in five cases. CONCLUSION: Tubal thickening, enlargement of ovarian stroma as reflected by the corrected cross-sectional area, and hyperintensity of this stroma on T2 weighted images probably related to edema were useful findings in these viable torsions. PMID- 15332254 TI - Uterine peristalsis: comparison of transvaginal ultrasound and two different sequences of cine MR imaging. AB - PURPOSE: To compare uterine peristalsis as seen on two different magnetic resonance (MR) imaging sequences and transvaginal ultrasound (TVUS), so as to better determine the best method for evaluating uterine peristalsis. MATERIALS AND METHODS: Eleven women in the periovulatory phase of the menstrual cycle underwent TVUS and cine MR imaging within a time period of three hours. Findings on cine MR images obtained with a serial 60 half-Fourier acquisition single-shot turbo spin-echo (HASTE) sequence and a 300 true fast imaging with steady-state precession (FISP) sequence were compared with TVUS. RESULTS: The image quality of the HASTE technique was the best among the three methods, followed by TVUS, and then true FISP technique (P < 0.001). Uterine peristalsis was detected in 100% of subjects using HASTE, 82% with True FISP, and 100% with TVUS. With HASTE technique, true FISP, and TVUS, respectively, endometrial stripping movement was identified in 100%, 82%, and 100%; direction was identified in 100%, 45% (P = 0.014), and 73%; and wave conduction toward the outer myometrium was identified in 55%, 9% (P = 0.025), and 0% (P = 0.014). CONCLUSION: Cine MR imaging by HASTE technique with a time resolution of two seconds best delineated uterine peristalsis compared to that by true FISP technique or with TVUS. PMID- 15332255 TI - Adaptive subtraction as an aid in MR-guided placement of catheters and guidewires. AB - PURPOSE: To demonstrate the utility of mask subtraction optimization in magnetic resonance (MR)-guided placement of catheters and guidewires. MATERIALS AND METHODS: MR-guided positioning of magnetically prepared catheters and guidewires was done by dynamically imaging a single thick slab at two frames per second. Selective visualization of the prepared parts of the devices was achieved by the use of a conventional baseline subtraction technique and by the use of an adaptive subtraction technique. In the latter, the best reference image is automatically selected from a fixed or a sliding subset of hitherto acquired dynamic images. The efficacy of both approaches was compared by tracking experiments in a flow phantom and in the aortoiliac arteries of a pig. RESULTS: Baseline subtraction produced adequate visualization of paramagnetic markers in the absence of subject motion and for fixed scan conditions. The sensitivity to subject motion and interactive modification of the scan parameters was greatly reduced by using adaptive subtraction. Adaptive subtraction images, other than conventional subtraction images, appeared to be insensitive to slow periodic motion, e.g., respiratory motion, and were only transiently affected by gross subject motion and interactive alterations of the scan parameters. CONCLUSION: Adaptive subtraction is superior to baseline subtraction for guiding the manipulation of catheters and guidewires in the presence of gross and periodic subject motion and whenever scan parameters are modified in the course of a procedure. PMID- 15332256 TI - Radiofrequency thermal ablation: correlation of hyperacute MR lesion images with tissue response. AB - PURPOSE: To investigate the hypothesis that the outer boundary of the hyperintense region observed in hyperacute (several minutes post-ablation) T2 and gadolinium contrast-enhanced (CE) T1-weighted magnetic resonance (MR) lesion images is an accurate predictor of eventual cell death from radiofrequency (RF) thermal ablation. MATERIALS AND METHODS: A low-field, open MR imaging system was used to guide an ablation electrode into a thigh muscle of five rabbits and acquire in vivo T2 and CE T1-weighted MR volumes. Ablation occurred by applying RF current for two minutes with the electrode's temperature maintained at 90 degrees +/- 2 degrees C. After fixation, we sliced and photographed the tissue at 3 mm intervals, using a specially designed apparatus, to obtain a volume of tissue images. Digital images of hematoxylin and eosin (H&E) and Masson trichrome stained histologic samples were obtained, and distinct regions of tissue damage were labeled using a video microscopy system. After the MR and histology images were aligned using a three-dimensional registration method, we compared tissue damage boundaries identified in histology with boundaries marked in MR images. RESULTS: The lesions have distinct zones of tissue damage histologically: a central zone of necrotic cells surrounded by an outer zone with cells that appeared non-viable and associated with marked interstitial edema. In 14 histology images from five lesions, the inner and outer boundaries of the outer zone were compared with the boundaries of a hyperintense rim that surrounds a central hypointense region in the T2 and CE T1-weighted MR images. For T2 and CE T1-weighted MR images, respectively, the mean absolute distance was 1.04 +/- 0.30 mm (mean +/- SD) and 1.00 +/- 0.34 mm for the inner boundaries, and 0.96 +/- 0.34 mm and 0.94 +/- 0.44 mm for the outer boundaries. The mean absolute distances for T2 and CE T1-weighted MR images were not sufficiently different to achieve statistical significance (P = 0.745, 0.818, for the inner and outer boundary, respectively). CONCLUSION: In hyperacute T2 and CE T1-weighted MR lesion images, observations strongly suggest that the outer boundary of the hyperintense rim corresponds to the region of eventual cell necrosis within a distance comparable to our ability to measure. This is good evidence that during RF ablation procedures, MR lesion images can be used to accurately localize the zone of irreversible tissue damage at the lesion margin. PMID- 15332257 TI - Quantitative evaluation of metal artifact reduction techniques. AB - PURPOSE: To develop a technique to quantify artifact, and to use it to compare the effectiveness of several approaches to metal artifact reduction, including view angle tilting and increasing the slice select and image bandwidths (BWs), in terms of metal artifact reduction, noise, and blur. MATERIALS AND METHODS: Nonmetallic replicas of two metal implants (stainless steel and titanium/chromium cobalt femoral prostheses) were fabricated from wax, and MR images were obtained of each component immersed in water. The differences between the images of each metal prosthesis and its wax counterpart were measured. The contributions from noise and blur were isolated, resulting in a measure of the metal artifact. Several off-resonance artifact reduction techniques were assessed in terms of metal artifact reduction capability, as well as signal to noise ratio and blur. RESULTS: Increasing the image BW from +/-16 kHz to +/-64 kHz was found to reduce the artifact by an average of 60%, while employing view angle tilting (VAT) alone was found to reduce the artifact by an average of 63%. The metal artifact reduction sequence (MARS), which combines several susceptibility artifact reduction techniques, resulted in the least amount of image distortion, reducing the artifact by an average of 79%. CONCLUSION: The results indicate that while VAT alone (with an image BW of +/-16 kHz) resulted in the smallest amount of total energy and no reduction in the signal-to-noise ratio compared to a conventional spin-echo pulse sequence, MARS resulted in significantly less artifact and dramatically less blur. PMID- 15332258 TI - Metabolites in ventricular cerebrospinal fluid detected by proton magnetic resonance spectroscopic imaging. AB - Normally, ventricular cerebrospinal fluid (CSF) contains low levels of all metabolite signals on proton magnetic resonance spectroscopic imaging (MRSI). We present here three cases (two with seizure disorders, one with a central nervous system lymphoma) who presented with unusually elevated CSF signals on MRSI. Based on chemical shifts and in vitro studies (in one case), the signals were assigned to propan-1,2-diol (PD), acetone, and lactate, respectively. These compounds were either exclusively, or more readily, detected in CSF than in brain. Proton MRSI conveniently screens both brain and CSF for abnormal metabolism simultaneously. PMID- 15332259 TI - Inflammatory pseudotumor of the liver: ferumoxide-enhanced MR imaging as a tiebreaker. AB - We examined a 70-year-old male patient with an inflammatory pseudotumor of the liver mimicking a peripheral-type cholangiocellular carcinoma. Ferumoxide enhanced magnetic resonance (MR) imaging revealed residual Kupffer cell function in liver parenchyma in and surrounding the inflammatory pseudotumor involvement, which suggested the diagnosis of inflammatory pseudotumor of the liver. We correlate the MR imaging and pathologic findings in this report. PMID- 15332260 TI - Parallel acquisition techniques in cardiac cine magnetic resonance imaging using TrueFISP sequences: comparison of image quality and artifacts. AB - PURPOSE: To compare image quality, artifacts, and signal-to-noise ratio (SNR) in cardiac cine TrueFISP magnetic resonance imaging (MRI) with and without parallel acquisition techniques (PAT). MATERIALS AND METHODS: MRI was performed in 16 subjects with a TrueFISP sequence (1.5 T; Magnetom Sonata, Siemens): TR, 3.0 msec; TE, 1.5 msec; flip angle (FA), 60 degrees. Three axes were scanned without PAT (no PAT) and using the generalized autocalibrating partially parallel acquisition (GRAPPA) and modified sensitivity encoding (mSENSE) reconstruction algorithms with an autocalibration mode to reduce scan time. A conventional spine array and a body flex array were used. Artifacts, image noise, and overall image quality were classified on a 4-point scale by an observer blinded to the implemented technique; for quantitative comparison, SNR was measured. RESULTS: With a PAT factor of two, acquisition time could be reduced by 39%. No PAT did not show artifacts, and GRAPPA revealed fewer artifacts than mSENSE. PAT provided inferior-quality scores concerning image noise and overall image quality. In quantitative measurements, GRAPPA and mSENSE (20.1 +/- 6.2 and 15.6 +/- 6.2, respectively) yielded lower SNR than no PAT (30.6 +/- 20.1; P < 0.05) and P < 0.001). CONCLUSION: Time savings in PAT are accompanied by artifacts and an increase in image noise. The GRAPPA algorithm was superior to mSENSE concerning image quality, noise, and SNR. PMID- 15332261 TI - Polyamine dendrimer-based MRI contrast agents for functional kidney imaging to diagnose acute renal failure. AB - PURPOSE: To choose an efficacious renal functional MRI contrast agent to image early renal tubular damage. We synthesized and compared smaller polyamine dendrimer-based MRI contrast agents (<60 kD) that, unlike Gd-[DTPA], transiently accumulate in renal tubules and can be used to visualize renal structural and functional damage. MATERIALS AND METHODS: Six dendrimer-based MRI contrast agents smaller than 60 kD were studied by high resolution dynamic micro-MRI and compared to Gd-[DTPA]-dimeglumine and Gadomer-17. The best agent, DAB-G2, was further tested in a mouse ischemia/reperfusion model to validate its efficacy. RESULTS: Despite unequal renal clearance rates, all polyamine dendrimer agents visualized the renal functional anatomy of the mice better than Gd-[DTPA]-dimeglumine and Gadomer-17. DAB-G2 was excreted most rapidly, yet was able to visualize mild renal tubular injury very early after injury. CONCLUSION: DAB-G2 was found to be the best candidate for functional kidney imaging and enabled early diagnosis of acute renal injury. PMID- 15332262 TI - Correlation of T1rho with fixed charge density in cartilage. AB - PURPOSE: To establish the specificity of T1rho with respect to fixed charge density (FCD) as a measure of proteoglycan (PG) content in cartilage during the onset of osteoarthritis (OA). MATERIALS AND METHODS: T1rho-weighted and sodium MRI were performed on cartilage samples of enzymatically degraded bovine explants and natural osteoarthritic human samples representing controlled and physiological models of OA, respectively. Spatial maps of T1rho and FCD (measured using the previously validated method of sodium MRI) were calculated from image data. Data were extracted from the maps and subjected to linear regression to compare changes in T1rho with changes in FCD in each model. Tissue samples were subjected to histological staining for a reduction in PG content. RESULTS: Plots of normalized T1rho rate vs. FCD were found to be strongly correlated (R2 > 0.75 and 0.85) in both models with nearly the same slope of approximately 1/2 (P > 0.51). Loss of PG in bovine and human tissue was confirmed by histology. CONCLUSION: The strong correlation of the FCD and T1rho data in both the controlled and physiological models demonstrates that changes in T1rho are due predominantly to changes in PG content. This work is a first step in establishing T1rho as a method of quantifying PG changes in early-stage OA. PMID- 15332263 TI - Weight-bearing MRI of patellofemoral joint cartilage contact area. AB - PURPOSE: To measure contact area of cartilage in the patellofemoral joint during weight bearing using an open MRI scanner. MATERIALS AND METHODS: We developed an MR-compatible back support that allows three-dimensional imaging of the patellofemoral cartilage under physiologic weight-bearing conditions with negligible motion artifact in an open MRI scanner. To measure contact areas, we trained observers using a phantom of known area and tested intra- and interobserver variability. We measured in vivo contact areas between the patella and femoral cartilage with the knee in 30 degrees of flexion, loaded and unloaded, in six volunteers. RESULTS: We were able to measure the contact area of the patellofemoral cartilage with small interobserver (CV 7.0%) and intraobserver (CV 3.0%) variation. At 30 degrees of knee flexion, mean contact area increased from 400 mm2 (unloaded) to 522 mm2(loaded to 0.45 times body weight per leg). CONCLUSION: Using an open magnet and specially designed apparatus, it is possible to image the patellar cartilage during physiologic loading. Knowledge of patellar cartilage contact area is needed to assess patellofemoral stress, which may be increased in patients with patellofemoral pain syndrome. PMID- 15332264 TI - New FOCI pulses with reduced radiofrequency power requirements. AB - PURPOSE: To propose new frequency offset corrected inversion (FOCI) pulses with significantly reduced radiofrequency (RF) power deposition for spin echo imaging by incorporating the variable-rate selective excitation (VERSE) schemes into the pulse design. MATERIALS AND METHODS: Two schemes are proposed to design the new FOCI pulses with dramatically reduced peak RF power requirements. In scheme A, the time-dilation function is derived from a predefined adiabaticity factor modulation function. In scheme B, the time-dilation function is predefined, while the adiabaticity factor is conserved. RESULTS: The new FOCI pulses are shown to be able to operate at reduced specific absorption rate (SAR), specifically at the same peak RF power as that of a five- or seven-lobe sinc inversion pulse of the same duration. Using the new FOCI pulse, significant gain in sensitivity was observed in in vivo spin-echo echo-planar imaging, which was attributed to the improved refocusing slice profile. CONCLUSION: The new FOCI pulses can replace the 180 degrees five- or seven-lobe sinc pulses in spin-echo imaging with the same peak RF power requirement and significantly improved slice profile. PMID- 15332265 TI - Temporal trends in the HIV-1 epidemic in Russia: predominance of subtype A. AB - During the period 1996-1997, three highly homogeneous variants of HIV-1 were identified, circulating among injecting drug users (IDUs) in the former Soviet Union republics. One of these belonged to HIV-1 genetic subtype A (IDU-A), another belonged to HIV-1 genetic subtype B (IDU-B) and the third was a recombinant between the first two variants (CRF03_AB). However, since 1997, the HIV-1 epidemic has affected an increasing number of geographic regions in Russia. This study was undertaken to survey the prevailing genetic variants and to estimate the current proportions of these three HIV-1 genetic subtypes in Russia. Blood samples were taken in 1999-2003 from 1090 HIV-infected individuals and analysed by gag/env HMA. The IDU-A variant was found to be the majority variant (89.7-100%) in 44 of 45 regions of the Russian Federation studied. The IDU-A variant was also found to spreading rapidly through heterosexual transmission in 1999-2003 (30/34, 88%). CRF03_AB predominates in the Kaliningrad region only (28/29, 96.6%). The IDU-B variant is currently of minor importance in the IDU epidemic but other European subtype B variants predominate among men having sex with men (18/18, 100%). Sequence analysis of the env V3 encoding regions derived from HIV-1 infected individuals in Yekaterinburg (the main centre of the HIV-1 epidemic in Russia in 2002-2003) showed that the IDU-A variant is still highly homogeneous. The mean pairwise nucleotide distance (n = 9) was 2.89 +/- 1.14 (range 1.36-6.14). However, the mean genetic distance between each sequence within the samples collected from the Yekaterinburg IDU-A variant subset and the IDU-A consensus is 2.51 +/- 1.06 (range 1.36-4.66) and considerably higher than in South Russia in 1996 (0.79 +/- 0.51, range 0.38-1.90). The current HIV-1 epidemic in Russia is almost entirely caused by a highly homogeneous A-subtype strain, which will influence vaccine development strategies and must be taken into account in the quality control of molecular tests for the diagnosis of HIV 1. PMID- 15332266 TI - High frequency of selection of K65R and Q151M mutations in HIV-2 infected patients receiving nucleoside reverse transcriptase inhibitors containing regimen. AB - The objective of the study was to determine retrospectively which substitutions in the reverse transcriptase (RT) gene are selected in vivo during nucleoside RT inhibitors (NRTI) containing regimen in HIV-2 infected subjects. Thirty-four HIV 2 patients having received NRTI-containing regimen with available specimens and amplifiable RT gene were studied. Analyses of RT gene were undertaken after a median NRTI exposure of 51 months (range: 5-128). Mutations at positions known to be involved in HIV-1 resistance were observed in 26/34 patients. Selection of Q151M mutation was observed in nine out of 34 isolates (26%) after a median NRTIs exposure of 41 months (range: 12-77). In 8/9 cases, Q151M mutation was associated with other substitutions at positions known to be involved in HIV-1 resistance: K65R (n = 6), D67N (n = 1), N69S or T (n = 2), K70R (n = 3), M184V (n = 4), S215Y (n = 1). Compared with HIV-1 infection, there is a high frequency of selection of Q151M mutation in HIV-2 infected patients receiving various combinations of NRTIs. In these highly thymidine analogue pretreated patients, the selection of thymidine analogue mutations was low suggesting that the pathway to resistance is very different between these two viruses. PMID- 15332267 TI - Comparison of serologic responses between Kaposi's sarcoma-positive and -negative men who were seropositive for both human herpesvirus 8 and human immunodeficiency virus. AB - Although the introduction of HAART decreased substantially the incidence of Kaposi's sarcoma (KS), KS remains the most common cancer among individuals infected with human immunodeficiency virus (HIV). To define markers for progression to KS from the asymptomatic infection of human herpesvirus 8 (HHV-8), serologic responses against HHV-8 were compared between KS-negative and -positive men who were seropositive for both HIV and HHV-8. There was no difference in prevalence of detectable neutralizing antibodies between the two groups. The prevalence of anti-ORF73 antibodies among the dual seropositive patients increased in proportion to their risk of KS. In specimens obtained from 11 HIV+ patients at different intervals over a period of 4-12 years, increase of anti ORF73 antibody titers was observed in the patients who developed KS but not in the patients who did not develop KS. These results suggest that there is a difference in serologic response against ORF73 between the HIV patients with and without KS. PMID- 15332268 TI - High prevalence of human T-lymphotropic virus type 1 (HTLV-1) in immigrant male to-female transsexual sex workers with HIV-1 infection. AB - Human T-lymphotropic virus type 1 and 2 (HTLV-1 and HTLV-2) infections in Europe are limited to intravenous drug users and migrants coming from areas in which they are endemic. A survey was undertaken of HTLV-1 and HTLV-2 infections in 393 recent immigrants: 167 HIV-1 positive subjects (including 52 male-to-female transsexual sex workers) and 226 pregnant HIV-1 negative women. The prevalence of HTLV-1 was 3.6% in the HIV-1 positive group and 0.9% in the HIV-1 negative group. The highest HTLV-1 prevalence in both groups was found in persons from Latin America, particularly those born in Peru (up to 26% in the HIV-1 positive group). All of the HIV-1/HTLV-1 co-infected individuals were male-to-female transsexual sex workers in whom the overall prevalence of HTLV-1 infection was 11.5%. HTLV-2 was only found in the HIV-1 positive group (prevalence 1.2%); all of the infected subjects were transsexual sex workers from Brazil (overall prevalence 6.4%). Phylogenetic analysis showed that all of the HTLV-1 isolates were of the cosmopolitan type, clustering with other strains circulating in the patients' birthplaces; the HTLV-2 isolates were of subtype 2a, and clustered significantly with other Brazilian strains. These results suggest the independent origin of each infection in the patient's birthplace. The data raise concerns about the further spread of HTLV infections mainly through the sexual route. PMID- 15332269 TI - Role of beauty treatment in the spread of parenterally transmitted hepatitis viruses in Italy. AB - The aims of the study were to evaluate the role of beauty treatments in the spread of acute viral hepatitis B (HBV) and acute viral hepatitis C (HCV) in Italy. Data from the surveillance system for acute viral hepatitis (SEIEVA) during the period 1997-2002 were used. After exclusion of subjects <15 years or >55 years old and reporting intravenous drug use or blood transfusion, the association of acute HBV and HCV cases with beauty treatments (tattooing, piercing, manicure/chiropody, and barber shop shaving) was estimated comparing 2,964 hepatitis B and 598 hepatitis C cases with 7,221 hepatitis A cases, used as controls, by multiple logistic regression analysis. The population attributable risk (PAR) to beauty treatments was estimated according to Levin's formula. Beauty treatments were associated with acute HBV (OR = 1.8; CI 95% = 1.5-2.1) and acute HCV (OR = 1.7; CI 95% = 1.2-2.3). The strongest association was found with barber shop shaving for HBV (OR = 1.8; CI 95% = 1.5-2.2) and with tattooing for HCV cases (OR = 5.6; CI 95% = 2.8-11.0). The estimates of the population attributable risk (PAR) indicate that nearly 15% of all acute HBV (17.4% in males) and 11.5% of all acute HCV cases (16.4% in males) occurring in 15-55 year old subjects not exposed to intravenous drugs or blood transfusion in Italy are due to beauty treatments. It is concluded that certain beauty treatments play an important role in the spread of HBV and HCV infections in Italy. PMID- 15332270 TI - Hepatitis B virus genotype D strains from Estonia share sequence similarity with strains from Siberia and may specify ayw4. AB - The genotypes and subtypes of 205 HBV isolates collected during 1989-2002 in Estonia and 14 other regions of the former USSR were determined by sequencing and phylogenetic analysis of the S gene. The in Europe prevailing genotypes, A and D, were also circulating in the whole territory of the former USSR including Estonia and accounted for 18.5 and 81% of the strains, respectively. All genotype A strains specified adw2, and a single genotype C strain specified adrq+. Most genotype D strains specified ayw3 and ayw2, although, three strains from Estonia and Siberia specified ayw4. Due to unique substitutions, Ser122 and Ala127, four strains could not be classified according to the subtype. One strain specifying ayw3 encoded Leu143 and Ala145 and was possibly an immune "escape" mutant. At phylogenetic analysis 93% of the Estonian genotype D strains belonged to a cluster specifying mainly ayw3 and were more similar to isolates from Siberia and the Far-East of Russia than to isolates originating from Central Russia which belonged to another cluster of strains specifying mainly ayw2. This pattern might be explained by part of the Estonian population, has roots east of European Russia, based on linguistic evidence. Eight dominant HBV strains represented by identical S gene sequences were identified, one within genotype A and seven within genotype D, three of which included isolates from Estonia and Siberia. Some of these strains were collected over a period of at least 13 years indicating there are genetically stable variants of HBV that remain conserved over decades. PMID- 15332271 TI - Characteristics of core promoter and precore stop codon mutants of hepatitis B virus in Vietnam. AB - In Asia, genotypes B and C are the most common genotypes of hepatitis B virus (HBV); and genotype C causes more severe liver disease. Core promoter/precore (CP/PC) mutants, known to be linked to these genotypes, could have an impact on the progression and severity of liver disease. Sera of 115 patients, including 39 acute and 76 chronic Vietnamese HBV infected patients, were tested for their liver profile, HBeAg, HBV genotypes, and HBV DNA level. Fragments of 282 nucleotides covering CP/PC were amplified, sequenced, and analysed. In the acute group, CP/PC mutants accounted for 38.4 and 25.6%, respectively. Genotype B was found to be predominant (74.3%, P < 0.05) and linked to the PC mutant (A1896) (P < 0.05). In the chronic group, CP/PC mutants accounted for 61.7 and 32.8%. CP mutants, especially the T1762/A1764 double mutant, were found to correlate with genotype C (81%, P < 0.001), liver cirrhosis, and hepatocellular carcinoma (P < 0.05). Therefore, genotype C in Vietnam, which carried high rate of C-1858 (70%), could play an important role in causing severe chronic liver disease. PMID- 15332272 TI - Evolution of precore/core promoter mutations in hepatitis B carriers with hepatitis B e antigen seroreversion. AB - The evolution of precore stop codon mutation (A1896) and dinucleotide mutation (T1762/A1764) in the basic core promoter (BCP) of hepatitis B virus (HBV) genome during transient seroconversion and seroreversion of hepatitis B e antigen (HBeAg) remains unclarified. Five HBeAg-positive HBV carriers who experienced transient seroconversion followed by seroreversion of HBeAg (Group I, 3.3%) and 3 HBeAg-negative HBV carriers with documented reversion of HBeAg (Group II, 2.5%) in a prospective cohort of 272 patients with chronic hepatitis B were thus identified. The sequential changes at the precore nucleotide 1896 and BCP dinucleotide 1762/1764 were determined by polymerase chain reaction and direct sequencing. At enrollement, precore A1896 and BCP T1762/A1764 were noted in 4 (50%) and 1 (13%) of the eight patients. During a median follow-up period of 58 months (range: 31-76 months), 12 episodes of transient HBeAg seroconversion followed by seroreversion were encountered in Group I patients and 3 episodes of HBeAg seroreversion in Group II patients. Accompanying acute exacerbations were found in two-thirds of patients with either HBeAg seroconversion or seroreversion. Overall, precore nucleotide A1896 remained identical in 73% and 83% of the seroconversion and seroreversion events, respectively. BCP dinucleotide T1762/A1764 remained unchanged in 94% and 92% of the seroconversion and seroreversion events, respectively. At the end of follow-up, only one had both precore and BCP mutations. In conclusion, these data suggested that HBeAg seroreversion might be due to the lack of sustained precore and BCP mutations after HBeAg seroconversion. Although uncommon, HBeAg seroreversion can be associated with hepatitis exacerbation. PMID- 15332273 TI - Molecular epidemiology of hepatitis C virus in Iran as reflected by phylogenetic analysis of the NS5B region. AB - Hepatitis C virus (HCV) subtypes were determined in 125 Iranian patients by phylogenetic analysis within the NS5B or 5'-UTR/core regions. Subtypes 1a and 3a were predominant accounting for 47 and 36%, whereas 1b and 4 accounted for 8 and 7%. This subtype distribution differs from that of Turkey and Pakistan, where subtypes 1b and 3a dominate and also from neighbouring Arabic countries where subtype 4 is the prevalent genotype. The Iranian 1a and 3a strains formed subclusters in the dendrogram indicating that these subtypes are indigenous to Iran. In contrast, the 1b strains intermixed with strains derived worldwide. Subtype 1a was frequent in South Iran (70%), while 3a was more prevalent in North West Iran (83%), a region with a high proportion of Turkish inhabitants. Patients infected by blood products had more frequently subtype 1a (57%), while younger drug users had more frequently subtype 3a (54%). Genotype 4 was over-represented among haemodialysis patients in Tehran. One strain, most similar to genotype 5, was highly divergent in the NS5B region and further analysis is needed to assess the systematic status of this strain. In half of the patients with unknown source of infection only the 5'-UTR could be amplified, most of which were from North West Iran and from patients younger than those with unknown source of infection with typable strains, mean age 29 versus 43 years. In conclusion, the NS5B sequence data revealed population based subtype patterns in Iran, the further study of which may help to understand the molecular epidemiology of HCV in a low endemic area. PMID- 15332274 TI - Major histocompatibility complex class-I presentation impaired in transgenic mice expressing hepatitis C virus structural proteins during dendritic cell maturation. AB - Hepatitis C virus (HCV) infection is often persistent, but its mechanism and pathogenesis remain unclear. One mechanism through which HCV escapes systemic immunosurveillance might be via impaired dendritic cells (DCs), which are the most potent type of antigen-presenting cells. We examined whether HCV causes immunosuppression in DCs during maturation. We isolated immature DCs from the bone marrow of two founder lineages of transgenic mice harboring HCV cDNA expressing HCV structural proteins (nucleotides 294-3435), and studied how DC function is modified by HCV expression. Our data showed that the capacity of DCs expressing HCV structural proteins to stimulate T-cells was significantly impaired. Moreover, the surface expression of major histocompatibility complex (MHC) class-I molecules was significantly impaired on infected DC, especially with respect to H-2D. The transportation of H-2D to the cell surface during DC maturation was inhibited by HCV expression. However, the total amount of H-2D molecules produced by DC expressing HCV was not impaired. These results indicated that the immune response of DC infected with HCV is diminished and might be associated with the mechanism of persistent HCV infection. PMID- 15332275 TI - Anti-HSV activity of lactoferrin and lactoferricin is dependent on the presence of heparan sulphate at the cell surface. AB - Lactoferrin (LF) is a multifunctional glycoprotein, which plays an important role in immune regulation and defense mechanisms against bacteria, fungi, and viruses. Lactoferricin (Lfcin) is a potent antimicrobial peptide generated from the N terminal part of LF by pepsin cleavage. In this study, we investigated the mechanisms of the anti-herpes simplex virus (anti-HSV) activity of LF and Lfcin. The results demonstrated that LF and Lfcin inhibited the entry of HSV into Vero cells. LF had no effect against HSV after the virus had entered the cells, while Lfcin exerted antiviral activity also after the initial binding of the virus to the host cell. The distribution of LF and Lfcin in the cells was investigated by immunogold-labeling and transmission electron microscope (TEM). LF was found mainly at the cell surface in cells expressing heparan sulphate. Lfcin was randomly distributed intracellularly. LF must be present at the cell surface to exert antiviral activity, while Lfcin exert its antiviral activity also when found mainly intracellularly. Both LF and Lfcin were dependent on the presence of heparan sulphate at the cell surface to exert their antiviral activity. PMID- 15332276 TI - Molecular testing for detection of in vitro infectivity of plasma pools contaminated with B19 virus. AB - B19 virus can be transmitted by contaminated blood or blood products. Recent observations, in healthy volunteers, suggest that active B19 infection can follow the administration of plasma pools with a concentration > or =10(7) genome equivalents/ml (geq/ml) of B19 DNA. However, patients receiving batches with levels of virus DNA lower than 10(4) geq/ml do not show any evidence of transmission of the virus. The aim of the study was to show, by in vitro assays, a threshold of viral load in B19 contaminated plasma pools over which the infection can be transmitted. Twenty plasma pools, each containing 960 single donations, were tested to correlate the viral load and the level of antibodies anti-B19 with the in vitro infectivity and expression of B19 virus. All the plasma pools, titrated for B19 viral load by competitive PCR, were inoculated into KU812Ep6 erythroid human cell line. Five of the nine contaminated plasma pools, with a B19 DNA concentration > or =3.60 x 10(6) geq/ml, were able to infect KU812Ep6 cells. In vitro infectivity was shown in KU812Ep6 cells at 24 h post-infection by in situ hybridisation and amplification assays for viral DNA and RNAs. Plasma pools with a viral load in the range of 6.00 x 10(3)-8.96 x 10(4) geq/ml did not show infectivity when inoculated into KU812Ep6 cells. Medium high titres of IgG antibodies anti-B19 were detectable in all the plasma pools and the neutralising activity associated with specific IgG anti-B19 may explain the lack of infectivity of plasma pools contaminated with a low viral load. In conclusion, in situ hybridisation and amplification assays for viral DNA and RNAs in KU812Ep6 cells inoculated with plasma pools can be valid assays to test for the presence of infectious virus in the production of B19-safe material. PMID- 15332277 TI - Epidemic resurgence of Chikungunya virus in democratic Republic of the Congo: identification of a new central African strain. AB - The resurgence of Chikungunya virus is described during an urban epidemic in Kinshasa Democratic Republic of the Congo, after 39 years without any isolation of the virus. Chikungunya virus was isolated in sera from nine patients with clinical symptoms. A 1,200 bp long partial sequence of the E1/3'UTR genomic region was determined for each isolate. All sequences clustered in the central African lineage. They constitute Chikungunya virus reference sequences for the Democratic Republic of the Congo. PMID- 15332278 TI - Coxsackievirus B3 replication and persistence in intestinal cells from mice infected orally and in the human CaCo-2 cell line. AB - Although the transmission of coxsackievirus B3 occurs mainly via the oral route, little is known about the primary replication and persistence of this agent in the intestine. To address this question, BALB/c mice were inoculated by gavage with coxsackievirus B3, Nancy strain. The mice were killed from 1 hr to 90 days after infection. The viral markers were detected in the small intestine using RT PCR, cell culture and detection of VP1 protein. Coxsackievirus B3 was detected positive by the three methods from hr 2 to day 45 after infection. By using monoclonal antibodies directed towards VP1, CD40 and CD26, the virus was shown to be present in the lymphocytes of the mucosa as soon as 2 hr after infection; in contrast, no virus was detected in the epithelial cells lining the intestinal lumen. Further experiments were performed to evaluate the capacity of coxsackievirus B3 to establish a persistent infection in two intestinal cell lines. In contrast to HT29 cells, the CaCo-2 cells were shown to develop a persistent infection for up to 20 passages, as demonstrated by the detection of viral RNA and VP1 protein. This study provides further evidence that, after infection by the oral route, the viral particles are concentrated in the lymphocytes of the mucosal layer. In addition, the results suggest that coxsackievirus B3 is capable of establishing a persistent infection in the small intestine that may act as a reservoir of viral particles for the delayed spread of the virus to other target organs. PMID- 15332279 TI - Coxsackie B viruses use multiple receptors to infect human cardiac cells. AB - Viral myocarditis is an inflammatory disease of the heart muscle that can be fatal. The primary viruses that have been linked to myocarditis and dilated cardiomyopathy are the human enteroviruses. The most common viruses associated with this disease are the Coxsackie B viruses and in particular Coxsackievirus B3 and Coxsackievirus B5. Early events in viral infection include attachment of the virus onto cell surface receptors. Even though, CD55 and Coxsackievirus adenovirus receptor protein (CAR) have been identified as receptors for Coxsackievirus B3, the exact mechanisms that Coxsackievirus B3 and B5 use to infect the cardiac muscle are not yet known. In this study, attempts were made to inhibit Coxsackievirus B3 and Coxsackievirus B5 infectivity of cardiac cells by using CAR and CD55 specific antibodies. The results show that these antibodies could not completely inhibit Coxsackievirus B3 and Coxsackievirus B5 binding or infectivity. Furthermore five new proteins have been identified that are used by Coxsackieviruses for binding to cardiac tissue and are distinct from CAR or CD55, leading us to believe that these viruses may use a different set of receptors for infection of cardiac muscle. PMID- 15332280 TI - Cross-species hybridization: characterization of gene expression in woodchuck liver using human membrane arrays. AB - Total RNA from normal adult woodchucks was analyzed using membrane arrays containing human cDNA clones, and the gene expression patterns were compared to human liver. Various hybridization and wash conditions were examined. In both the woodchuck and human livers, 352 genes were identified as highly expressed (Z scores > or =1.96). These genes represented numerous liver functions: transcription, RNA processing, signal transduction, protein synthesis and degradation, as well as enzymes. Several genes were selected and expression was verified by Northern blots for woodchuck liver. There were no false positives but 29 genes were identified as false negatives, expressed only in human liver. Possible reasons for these false negatives were the length and percentage of homology between the two species, differences in the distribution and types of mismatches, and the sequence region spotted on the array. These were assessed by examining expression of the transferrin gene in both species. A 200-fold range of RNA concentration (0.1-20 microg total RNA) was also examined and the optimal RNA concentration was determined to be 5 microg. Membranes were capable of being hybridized and reprobed at least five times. The study demonstrates that cross species hybridization is a valid method for identifying gene expression in woodchuck liver. PMID- 15332281 TI - Emergence and continued circulation of dengue-2 (genotype IV) virus strains in northern India. AB - Dengue (DEN) is an acute mosquito borne viral disease of mankind. Off late it has become an important public health concern in Southeast Asia. Although, all the four known dengue virus serotypes (DEN-1 to 4) are reported from time to time, in the recent past, DEN-2 has emerged as the predominant type, being the causative agent of several outbreaks of dengue fever (DF) and dengue haemorrhagic fever (DHF) in India. To elucidate the true molecular epidemiology of these viruses, we have sequenced C-prM gene junction (454 nucleotides) of 11 DEN-2 viruses directly from patient serum. The C-prM gene junction was amplified initially by reverse transcription-polymerase chain reaction followed by automated DNA sequencing. These sequences provide unique information with regard to molecular epidemiology when compared to other DEN-2 sequences from diverse geographic origins. The sequence analysis revealed that most of the mutations in this region remained silent, except a few at the carboxy-terminal of the capsid. Reported phylogenetic analysis classifies DEN-2 viruses into five distinct genotypes. The Gwalior DEN-2 viruses, included in the present study were classified into genotype-IV, and were found to be most closely related to Delhi 1996 DEN-2 viruses and FJ 10/11 strains prevalent in the Fujian state of China. However, two earlier Indian isolates of DEN-2 were classified into genotype-V. The present study indicates that genotype V of DEN-2 has been replaced by genotype IV during the past decade, which continues to circulate silently in north India, and have the potential to reemerge and cause major epidemics of DF and DHF. PMID- 15332282 TI - Human T lymphotropic virus types I and II proviral sequences in Argentinian blood donors with indeterminate Western blot patterns. AB - Human T-cell lymphotropic virus (HTLV) seroindeterminate blood donors have been reported worldwide including Argentina. To investigate the significance of HTLV I/II seroindeterminate Western blot (WB) patterns, we conducted an 8-year cross sectional study. Of 86,238 Argentinian blood donors, 146 sera were reactive by screening tests. The WB results indicated that 20% were HTLV-I reactive, 8% HTLV II reactive, 61% indeterminate, and 11% negative. The overall seroprevalence was 0.034% for HTLV-I, 0.014% for HTLV-II, and 0.103% for indeterminate. In 57 reactive specimens, HTLV-I/II provirus could be examined by type specific PCR for tax, pol, and env regions. When at least two gene fragments were amplified HTLV I/II infection was considered confirmed. PCR results confirmed all WB seropositive samples for HTLV-I (n = 15), and HTLV-II (n = 7), and the only WB negative case was also PCR negative, showing a complete concordance between PCR and WB. However, of 34 WB seroindeterminate sera studied by PCR, in 5 was proviral DNA amplified. According to our criteria PCR confirmed one to be HTLV-I, and one HTLV-II, 3 remained indeterminate since only tax sequences were amplified. Among WB indeterminate samples tested by PCR, most of their serological profile showed reactivity to gag codified proteins but lacked env reactivities (70%). One sample with a WB gag pattern showed proviral tax sequences, but of the four samples with reactivity to env proteins GD21 (n = 3) or rgp46II (n = 1) PCR results indicated that one was HTLV-I, one was HTLV-II, and two were indeterminate (only tax sequences). In conclusion, the majority of HTLV-seroindeterminate WB donors exhibited a gag indeterminate profile lacking HTLV provirus, and were thus considered uninfected. However, seroreactivity to env proteins, in particular to GD21, may indicate infection and a follow-up study of each seroreactive blood donor should be considered. PMID- 15332283 TI - Secretory IgA antibodies provide cross-protection against infection with different strains of influenza B virus. AB - This study examined whether secretory IgA (S-IgA) antibodies (Abs) could confer cross-protective immunity against infection with influenza B viruses of antigenically distinct lineages. Wild-type or polymeric Ig receptor (pIgR) knockout (KO) mice were immunized by infection with different B viruses or by intranasal (i.n.) administration with different inactivated vaccines. Four weeks later mice were challenged with either the B/Ibaraki/2/85 virus, representative of the B/Victoria/2/87 (B/Victoria)-lineage, or B/Yamagata/16/88 virus, representative of the B/Yamagata-lineage. Three days after challenge, nasal wash and serum specimens were assayed for IgA and IgG Abs specific for challenge viral antigens and for protection against challenge viruses. In wild-type mice, B/Ibaraki (or B/Yamagata) cross-reactive IgA Abs were detected at higher levels when infected or immunized with homologous-lineage viruses and at lower levels when infected or immunized with heterologous-lineage viruses. There was a correlation between the amount of nasal cross-reactive IgA Ab and the efficacy of cross-protection with a homologous-lineage virus. In mice lacking the pIgR, nasal cross-protective IgA Abs were only marginally detected in vaccinated mice and an accumulation of IgA in the serum was observed. This reduction of nasal IgA was accompanied by inefficient cross-protection against the B/Ibaraki (or B/Yamagata) virus infection. These results suggest that challenge viral-antigen cross reactive S-IgA in nasal secretions induced by i.n. infection or vaccination is involved in providing cross-protection against challenge infection with virus within either the B/Victoria- or B/Yamagata-lineage. PMID- 15332284 TI - Antigenic alteration of influenza B virus associated with loss of a glycosylation site due to host-cell adaptation. AB - Effects of host-cell adaptation of the hemagglutinin (HA) protein were evaluated by the analyses of four pairs of recent influenza B field isolates, each pair consisting of an Madin Darby canine kidney (MDCK)- and an embryonated chicken egg derived isolates from the same clinical specimen. Among the isolates examined, all of the MDCK-derived isolates retained glycosylation site at amino acid 197 on the HA1 molecule, whereas three egg-derived isolates lost it. Antigenic difference in the HA molecule between an MDCK- and an egg-derived isolates of three of these pairs was demonstrated to be associated with the glycosylation 197. Replication of the MDCK-derived isolates was suppressed in eggs, suggesting that the presence of the glycosylation 197 was disadvantageous to replication in eggs. Virus-binding affinity assay revealed that the loss of carbohydrate chain did not significantly alter the preferential recognition of sialic acid linkage. Immunogenicity and vaccine efficacy of an MDCK- and an egg-derived clones of B/Akita/27/2001, the former retained the glycosylation 197 and the latter lost it, were compared in a hamster model. When formalin-inactivated whole virion vaccines prepared from the paired isolates were administered into hamsters, no significant difference between them was observed in protective ability against challenges by the homologous and heterologous clones. Implication of the egg adaptation of influenza virus to antigenic surveillance of the field isolates as well as the selection of vaccine strains, and possibility of the involvement of the viral protein(s) other than the HA in the egg adaptation were discussed. PMID- 15332285 TI - Molecular epidemiology of respiratory syncytial virus in Kilifi district, Kenya. AB - Respiratory syncytial virus (RSV) causes significant burden of disease during infancy and childhood. This study examined the genetic relatedness of RSV positive samples from child inpatients and outpatients and a birth cohort from a rural coastal district of Kenya and also the distribution of strains between these three groups. Clinical samples were collected over a 4-year period in Kilifi District, Kenya from community and hospital surveillance. Three hundred ninety seven of 1,044 nasal specimens from children (under 5 years old) attending Kilifi District Hospital, and from community-monitored infants, were positive for RSV by multiplex RT-PCR. Of these, 376 samples were analysed further by restriction fragment length polymorphisms (RFLP) of the nucleocapsid (N) and attachment (G) protein genes. The G gene was sequenced for 109 samples and phylogenetic analysis carried out. The group A samples from Kilifi fell into two clusters based on G gene sequences, while only one group B cluster was observed. One RSV-B sample from 2003 demonstrated the presence of a 60-nucleotide duplication within the G gene, clustering with similar isolates from Buenos Aries from 1999. All had similar sequences to isolates from the UK, USA, Spain, or Uruguay. The Kilifi District samples showed greater than 97% homology to isolates from South Africa and Mozambique and 91-94% homology to isolates from The Gambia. Samples from different sources, clearly differing in disease severity, did not differ in genotype characteristics, suggesting that disease causing variants are a general reflection of infections within this community. PMID- 15332286 TI - Variable patterns of varicella-zoster virus reactivation in Ramsay Hunt syndrome. AB - The mechanism by which reactivation of varicella-zoster virus (VZV) causes facial paralysis in Ramsay Hunt syndrome remains unclear. The relationship between VZV load and the onset of facial paralysis was analyzed in 42 patients with Ramsay Hunt syndrome. The patients were divided into three groups according to the times of appearance of zoster and of facial paralysis; group I (zoster preceding, n = 13), group II (simultaneous, n = 22), group III (paralysis preceding, n = 7). A real-time quantitative PCR assay was used to measure VZV DNA copy number in saliva, and paired sera were assayed for anti-VZV IgG and IgM antibodies. In group I, the VZV DNA-positive rate was low and virus load decreased gradually after the initial hospital visit around the time of onset of paralysis. The level of anti-VZV antibodies had in most cases already increased at that time. In group III, viral load tended to increase after the onset of paralysis and peaked around the time of appearance of zoster. The level of anti-VZV antibodies was low at the onset of paralysis but showed a significant increase when paired sera were tested. In group II, virus load and changes in level of anti-VZV antibodies either resembled group I or group III behavior. These results indicate that facial paralysis in Ramsay Hunt syndrome can occur at various times between the early and the regression phase of VZV reactivation, suggesting that there are variable patterns of development of facial nerve dysfunction caused by VZV reactivation and the progression of neuritis. PMID- 15332287 TI - YMDD mutants in patients with chronic hepatitis B before treatment are not selected by lamivudine. AB - Hepatitis B virus (HBV) mutants with mutations in the YMDD motif of viral DNA polymerase/reverse transcriptase are described in patients infected with HBV who have not received lamivudine therapy, but their pathogenetic potential is not clear. These mutants were detected by the polymerase chain reaction with peptide nucleic acid clamping in pretreatment sera from two patients who later received lamivudine. One patient developed acute exacerbation with hepatic encephalopathy and received lamivudine along with plasma exchange, which were effective on his illness. YIDD mutants were detected in all three pretreatment sera and both posttreatment sera from him. HBV DNA clones from pretreatment and posttreatment sera, however, did not have the same amino acid sequence. In the other patient who developed severe breakthrough hepatitis after receiving lamivudine, YIDD mutants were detected in two pretreatment and two posttreatment sera. When amino acid sequences of HBV DNA clones with the YIDD mutation were compared before and after he received lamivudine, however, they were not in accord. Hence, YIDD mutants in both patients with chronic hepatitis B before treatment were not selected by lamivudine after they had been placed on it. Numerous amino acid conversions were detected in HBV DNA clones with YIDD mutations, and some of them created stop codons in the overlapping S gene sequence. In Conclusion, HBV mutants with mutations in the YMDD motif in patients before treatment would not be selected by lamivudine or induce breakthrough hepatitis, and some of these would not be replication-competent due to stop codons in the S gene. PMID- 15332288 TI - Analgesic mixtures containing phenacetin. PMID- 15332289 TI - Phenazopyridine hydrochloride. PMID- 15332290 TI - Phenolphthalein. PMID- 15332291 TI - Phenoxybenzamine hydrochloride. PMID- 15332292 TI - Phenytoin. PMID- 15332293 TI - Polybrominated biphenyls (PBBs). PMID- 15332294 TI - Polychlorinated biphenyls (PCBs). PMID- 15332295 TI - Alternative modalities for asthma that reduce stress and modify mood states: evidence for underlying psychobiologic mechanisms. AB - OBJECTIVE: To review the evidence supporting a role for psychological interventions in the treatment of asthma, with particular emphasis on underlying psychobiological mechanisms. DATA SOURCES: Independent literature searches on MEDLINE, PREMEDLINE, Cochrane Library, and PSYCHINFO from their respective inception to 2003 were performed. Separate searches were performed for psychological stress and asthma, psychoneuroimmunology and asthma, stress management, relaxation, asthma, complementary and alternative medicine and asthma, and immune function and psychological intervention. The search was not limited based on language of publication. STUDY SELECTION: Supportive evidence from overlapping research was included based on the expert opinion of the author and through discussions with consultants in the field. RESULTS: This review first discusses human and animal studies focused on psychological stress and the effects of stress on the neuroendocrine and immune system, emphasizing the implication of these effects on asthma. Second, studies that evaluated the influence of stress reduction modalities on neuroendocrine and immune function were examined. Existing evidence from human clinical studies that explored the role of psychological interventions for asthma is reexamined in this context. CONCLUSIONS: A growing appreciation of the interactions among behavioral, neural, endocrine, and immune processes suggests possible mechanisms through which psychological interventions for asthma may be operating. This review provides a framework in which we can begin to see links among these systems that might provide new insights to guide future explorations. PMID- 15332296 TI - Analysis of the polarization-dependent diffraction from a metallic grating by use of a three-dimensional combined vectorial method. AB - A three-dimensional combined vectorial method, which is based on the finite difference time-domain algorithm and vectorial diffraction formulation, is introduced to analyze the interaction of a realistic focused beam with a metallic grating in an optical storage system. The diffracted field patterns and the detected signals are calculated for gratings with different geometries, and the polarization-dependent characteristics are studied. The combined method can give accurately the detected signals and the field pattern at any desired position of the optical storage system. PMID- 15332297 TI - Bisacridines with aromatic linking chains. Synthesis, DNA interaction, and antitumor activity. AB - Synthesis of a series of bisacridine derivatives containing rigid aromatic linking chains is described. Their DNA interaction and in vitro cytotoxicity against HT-29 human carcinoma cells are reported. PMID- 15332298 TI - Bibliography. Current world literature. Kidney cancer. PMID- 15332299 TI - Interview with Dr Franck Renouard, Chairman of the 13th Annual EAO Scientific Meeting. Interview by Carlos Aparicio. PMID- 15332300 TI - [Comment about "Who pays for the impact?"]. PMID- 15332301 TI - Stressing the virtues of the immune system. PMID- 15332302 TI - Mitochondrial permeability: dual role for the ADP/ATP translocator? AB - The ADP/ATP translocator (or adenine nucleotide translocase; ANT) is thought to play a dual role: in the transport of ADP and ATP across the mitochondrial inner membrane and in the formation of the mitochondrial permeability-transition pore (mtPTP), a nonspecific pore that is an important mediator of apoptosis (programmed cell death). However, Kokoszka et al. have shown that mitochondria from livers of 'ANT-knockout' mice, in which the ANT has been genetically inactivated, still possess mtPTP activity. From this, the authors conclude that the ANT is a non-essential component of the mtPTP that may be dispensable for mtPTP-associated cell death. These results, which contradict previous evidence and cast doubt on a widely accepted model for the mtPTP (ref. 1), warrant scrutiny and call for a fundamental reappraisal of the role of the ANT in liver metabolism. PMID- 15332303 TI - Bibliography. Current world literature. Reproductive endocrinology. PMID- 15332304 TI - Exploitation of host cell cytoskeleton and signalling during Listeria monocytogenes entry into mammalian cells. AB - Deciphering how Listeria monocytogenes exploits the host cell machinery to invade mammalian cells during infection isa key issue for the understanding how this food-borne pathogen causes a pleiotropic disease ranging from gastro-enteritis to meningitis and abortions. Using multidisciplinary approaches, essentially combining bacterial genetics and cell biology, we have identified two bacterial proteins critical for entry into target cells, InlA and InlB. Their cellular ligands have been also identified: InlA interacts with the adhesion molecule E cadherin, while InlB interacts with the receptor for the globular head of the complement factor Clq (gClq-R), with the hepatocyte growth factor receptor (c Met) and with glycosaminoglycans(including heparan sulphate). The dynamic interaction between these cellular receptors and the actin cytoskeleton is currently under investigation. Several intracellular molecules have been recognized as key effectors for Listeria entry into target cells,including catenins (implicated in the connection of E-cadherin to actin) and the actin depolymerising factor/cofilin (involved in the rearrangement of the cytoskeleton in the InlB-dependent internalisation pathway). At the organism level, species specificity has been discovered concerning the interaction between InlA and E cadherin, leading to the generation of transgenic mice expressing the human E cadherin, in which the critical role of InlA in the crossing of the intestinal barrier has been clearly determined. Listeria appears as an instrumental model for addressing critical questions concerning both the complex process of bacterial pathogenesis and also fundamental molecular processes, such as phagocytosis. PMID- 15332305 TI - Republication of the article "Exploitation of host cell cytoskeleton and signalling during Listeria monocytogenes entry into mammalian cells". PMID- 15332306 TI - Waiver of consent for emergency research. PMID- 15332307 TI - The effect of emergency department crowding on ambulance availability. PMID- 15332308 TI - Contrast-induced thyrotoxicosis. PMID- 15332309 TI - Direct humeral head manipulation (Oza maneuver) for anterior shoulder dislocations. PMID- 15332310 TI - Ultrasonographic diagnosis of pleural effusion in congestive heart failure. PMID- 15332311 TI - Effect of Ramadan on surgical emergencies. PMID- 15332312 TI - Creating lifetime images of health and disease. PMID- 15332313 TI - [Patient expectations before elective surgery of inguinal hernia. What are the sequelae for ambulatory surgery of inguinal hernia with reference to DRG?]. PMID- 15332315 TI - Standard requirements for the preparation of manuscripts submitted for publication in the journal Roczniki Panstwowego Zakladu Higieny. PMID- 15332314 TI - [Management of gigantic recurrent incisional hernia. Repair by modified Ramirez component separation technique and a skin sliding flap]. PMID- 15332316 TI - National Advocacy and President's Forum. PMID- 15332317 TI - CPR--making sensible end-of-life choices. PMID- 15332318 TI - Low dose vasopressin for septic shock: where does it belong in therapy? PMID- 15332319 TI - Use your own best judgment. PMID- 15332320 TI - On beauty... PMID- 15332321 TI - Pancreas transplantation. AB - Over the last 15 years whole organ pancreas transplantation has emerged as the treatment of choice for selected patients with uremia and Type I Diabetes Mellitus. Improvements in surgical technique, better understanding of transplant related complications and advances in immunosuppressive therapy have encouraged the application of this procedure to an increasing number of patients. Pancreas transplantation occurs under three primary scenarios: simultaneous kidney pancreas transplantation, pancreas transplantation after kidney transplantation, and pancreas transplant alone. Overall results are excellent with 90%-95% one year patient survival, and 85%-90% of patients achieving normal glycemic control. There also exists a significant long-term survival advantage among the simultaneous kidney pancreas transplant group. PMID- 15332322 TI - Accumulation of boron in human malignant glioma cells in vitro is cell type dependent. AB - It has been shown that human malignant glioma tumours consist of several subpopulations of tumour cells. Due to heterogeneity and different degrees of vascularisation cell subpopulations possess varying resistance to chemo- or radiation therapy. Therefore, therapy is dependent on the ability to specifically target a tumour cell. Boron neutron capture therapy (BNCT) is a bimodal method, in radiation therapy, taking advantage of the ability of the stable isotope boron 10 to capture neutrons. It results in disintegration products depositing large amounts of energy within a short length, approximately one cell diameter. Thereby, selective irradiation of a target cell may be accomplished if a sufficient amount of boron has been accumulated and hence the cell-associated boron concentration is of critical importance. The accumulation of boron, boronophenylalanine (BPA), was investigated in two human glioma cell subpopulations and a human fibroblast cell line in vitro. The cells were incubated at low boron concentrations (0-5 microg B/ml). Oil filtration was then used for separation of extracellular and cell-associated boron. Inductively coupled plasma atomic emission spectroscopy (ICP-AES) was used for boron determination. Significant (P < 0.05) differences in accumulation ratio (relation between cell-associated and extracellular boron concentration) between human malignant glioma cell lines were found. Human fibroblasts, used to represent normal cells, showed a growth-dependent uptake and a lower accumulation ratio than the glioma cells. Our findings indicate that BPA concentration, incubation time and differences in boron uptake between cell subpopulations should be considered in BNCT. PMID- 15332323 TI - Volume reconstruction techniques improve the correlation between histological and in vivo tumor volume measurements in mouse models of human gliomas. AB - Assessment of therapy efficacy using animal models of tumorigenic cancer requires the ability to accurately measure changes in tumor volume over the duration of disease course. In order to be meaningful, in vivo tumor volume measurements by non-invasive techniques must correlate with tumor volume measurements from endpoint histological analysis. Tumor volume is frequently assessed by endpoint histological analyses approximating the tumor volume with geometric primitives such as spheroids and ellipsoids. In this study we investigated alternative techniques for quantifying histological volume measurements of tumors in a xenograft orthotopic mouse model of human glioblastoma multiforme, and compared these to in vivo tumor volume measurements based on magnetic resonance imaging (MRI) data. Two techniques leveraging three-dimensional (3D) image analysis methods were investigated. The first technique involves the reconstruction of a smoothed polygonal model representing the tumor volume from histological section images and is intended for accuracy and qualitative assessment of tumor burden by visualization, while a second technique which approximates the tumor volume as a series of slabs is presented as an abbreviated process intended to produce quantitatively similar volume measurements with a minimum of effort required on behalf of the investigator. New software (QuickVol) designed for use in the first technique, is also discussed. In cases where tumor growth is asymmetric and invasive, we found that 3D analysis techniques using histological section images produced volume measurements more consistent with in vivo volume measurements based on MRI data, than approximation of tumor volume using geometric primitives. Visualizations of the volumes represented by each of these techniques qualitatively support this finding, and suggest that future research using mouse models of glioblastoma multiforme (genetically engineered or xenograft) will benefit from the use of these or similar alternative tumor volume measurement techniques. PMID- 15332324 TI - Inducible expression of p57KIP2 inhibits glioma cell motility and invasion. AB - To examine the role of p57KIP2 in human malignant glioma cells, we studied its expression in a panel of human malignant glioma specimens by western blot and immunohistochemical analysis. To determine the effects of p57KIP2 expression on the phenotype of glioma cells, we analyzed two inducible stably transfected p57KIP2 expressing glioma cell lines. Expression of p57KIP2 was induced in U373 and U87 malignant glioma cells with doxycycline using the tetracycline repressor system. A phagokinetic track assay on gold particles was used to investigate differences in cell migration between p57KIP2 expressing and non-expressing control cells. The effects of the extracellular matrix (ECM) on U373 motility was determined in p57+ and p57-cells on surfaces coated with 5 microg/cm2 of fibronectin, laminin, type I and type IV collagens. The invasion of p57+ and p57- glioma cells across BD Biocoat Matrigel invasion chambers was then determined. p57KIP2 was weakly expressed in 4/6 glioblastoma (GBM) specimens by western blot. By immunohistochemistry, p57KIP2 immunoreactivity was positive in 8/40 GBMs, and was primarily nuclear in location. The motility of U373 glioma cells was significantly reduced after p57KIP2 induction. The presence of ECM proteins did not further alter the motility of p57+ and p57- glioma cells. The results of the invasion chamber assay showed that p57+ cells exhibited a 35% reduction in their invasive capacity as compared to p57- cells. These data suggest that p57KIP2 is expressed in at least some malignant gliomas. Inducible expression of 57KIP2 in cell lines deficient in this cyclin-dependent kinase inhibitor reduces their otility and invasiveness. PMID- 15332325 TI - Intralesional mitoxantrone biopolymer-mediated chemotherapy prolongs survival in rats with experimental brain tumors. AB - The present study was designed to test the efficacy of intratumoral biopolymer mediated mitoxantrone chemotherapy in the rat brain 9L glioma model. Mitoxantrone polymers were tested in vitro in 9L and C6 cell cultures for 10 days. Subsequently, adult Fisher 344 rats were implanted with 5 x 10(4) 9L glioma cells in the frontal region of the brain. In a first experiment, 2 days after cells inoculation, one group of rats were implanted with a biopolymer loaded with 4 mg of mitoxantrone at the tumor site. A second group of rats received drug-free biopolymers and served as controls. In a second experiment, rats were implanted with a biopolymer loaded with 2 mg of mitoxantrone. Another group of rats received 2 mg of mitoxantrone intraperitoneally. Controls received drug-free biopolymers. Rats were sacrificed as soon as they developed progressive neurological deficits. In the first experiment mean survival of mitoxantrone treated rats was 10+/-2 vs. 15+/-2 days for the control group (P = 0.0003). Early morbidity was seen in 60%, and impaired wound healing was seen in 40% of the 4 mg mitoxantrone treated animals. In the second experiment mean survival of mitoxantrone-treated rats was significantly longer than that of the control group (P < 0.0001) with 33+/-7 vs. 13.8+/-2 days for the control group. Only transient early morbidity (20%) was observed at this dose. All rats in the intraperitoneally mitoxantrone-treated group died within the first 4 days after injection. We conclude that controlled-release EVAc carriers deliver biologically active mitoxantrone in a sustained fashion. In vivo biopolymer-mediated mitoxantrone in loco chemotherapy can significantly prolong survival in rats with intracerebral 9L gliomas. Morbidity is mainly dose related, and can be reduced at acceptable levels without compromising the therapeutic effect. PMID- 15332327 TI - Neurolymphomatosis. PMID- 15332326 TI - BCNU down-regulates anti-apoptotic proteins bcl-xL and Bcl-2 in association with cell death in oligodendroglioma-derived cells. AB - Oligodendroglial differentiation in gliomas is associated with enhanced sensitivity to chemotherapy. Antiapoptotic proteins, Bcl-xL and Bcl-2, are over expressed in early passage cell lines derived from glioblastomas. Down-regulation of Bcl-xL and Bcl-2 with DNA antisense oligonucleotides promotes cell death in glioblastoma cells. Changes of expression of Bcl-xL and Bcl-2 after chemotherapy treatment have not been studied in glioma subtypes. The current experiments correlate decreased expression of both Bcl-xL and Bcl-2 after BCNU chemotherapy and cell death in two oligodendroglioma-derived cell lines. Expression of Bcl-2 family member proteins Bcl-xL, Bcl-2, and Bax were assessed in glioma cells both before and after chemotherapy treatment. Cell survival was assessed with a crystal violet bioassay. Levels of expression of Bcl-2 and Bcl-xL were elevated in two early passage oligodendroglioma-derived cell lines compared with a non neoplastic glial cell line. BCNU chemotherapy markedly down-regulated expression of Bcl-xL and Bcl-2 proteins in both oligodendroglioma-derived cell lines. Changes in expression of Bcl-xL and Bcl-2 were associated with the increased sensitivity to chemotherapy. There were no changes noted in expression of Bax after BCNU treatment. Modulation of expression of the anti-apoptotic proteins, Bcl-xL and Bcl-2, in the two oligodendroglioma-derived cell lines was associated with increased sensitivity to BCNU chemotherapy. Down-regulation of Bcl-xL and Bcl-2 resulted in reversal of the ratio of Bax/Bcl-xL and Bax/Bcl-2 and enhanced cell death after treatment with BCNU. Mechanisms that control expression of the anti-apoptotic proteins Bcl-xL and/or Bcl-2 may be effective targets in treatment strategies in patients with malignant gliomas. PMID- 15332328 TI - Case report: Pseudomonas aeruginosa-related intervertebral discitis in a young boy with medulloblastoma. AB - We report a case of a 15-year-old boy with desmoplastic medulloblastoma of the posterior fossa (T3M3, according to Chang classification) incompletely resected, with leptomeningeal and nodular spread in the posterior fossa and in the cervical and thoracic tracts of the spine, treated with sequential high dose iv chemotherapy and with hyperfractionated cranio-spinal radiotherapy. While on maintenance chemotherapy, the boy developed fever and septic status caused by Pseudomonas aeruginosa, and 1 week later also low back pain. Magnetic resonance imaging (MRI) demonstrated abnormal signal in the fourth ventricle and in the dorso-lumbar tract suggesting medulloblastoma recurrence, so he started with a chemotherapy program. Due to a worsening of back pain, a second MRI of the spine was performed that showed a spondilodiscitis of T11-T12 and L1-L2 discs. The histological and cultural examination of a fine-needle biopsy of the L1-L2 disc revealed the presence of P. aeruginosa. So patient was treated with intensive antibiotic therapy with resolution of the infection. Spondilodiscitis is a rare complication in neoplastic patients, maybe due to either immunodeficient status or invasive procedures such as lumbar puncture. This case demonstrates that MRI is a useful method for differentiating between infection and malignancy in the spine, but sometimes it may be difficult to distinguish metastatic tumor from a lesion due to spondilodiscitis. In this case surgicopathological assessment is crucial and mandatory. PMID- 15332329 TI - Intracranial and spinal melanotic schwannoma in the same patient. AB - Melanotic schwannoma is a nerve sheath tumor composed of melanin - producing cells with ultrastructural features of Schwann cells, which is very rare. These tumors can appear intracranially and less common in the spinal canal. The prognosis is usually good although cases with metastases and death are described in the literature. We present the unusual and rare case of a young man with a spinal melanotic schwannoma at L5/S1 who presented 30 months later with an intracranial melanotic schwannoma. PMID- 15332330 TI - Multivariate analysis of clinical prognostic factors in children with intracranial ependymomas. AB - The optimal postoperative management of pediatric intracranial ependymomas is controversial. We analyzed clinical prognostic factors for their influence on outcome in such children. Our retrospective series included 15 with supratentorial and 28 with infratentorial tumors. Twenty ependymomas were grade II, and 23 were anaplastic. Complete resection was performed in 18 patients, incomplete resection in 19, and stereotactic biopsy in 6. Radiotherapy was done in 31 patients and chemotherapy in 13. The surviving patients have been followed 8-232 months (median: 69 months). The median survival time was 30 months, and 5 year overall survival and progression-free survival rates were 53.9% and 45.9%, respectively. By tumor site: supratentorial, 56.6% and 50.9%; infratentorial, 52.3% and 42.5%. Multivariate analysis identified complete resection (5-year progression-free survival, 71.8%) and age <3 years old as significant favorable and adverse prognostic features (relative risk, 2.59; 95% CI, 1.05-6.38), respectively. Twenty-six children relapsed 1-107 months after diagnosis (median: 12 months). Relapses were local in 22 cases, and combined local and distant in three cases. Only one of 15 patients with supratentorial tumors developed isolated spinal metastasis. Failure at the primary site is the major obstacle to improve cure rates. The extent of surgical resection and age were the only statistically significant prognostic factors. PMID- 15332332 TI - Correlation of clinical features and methylation status of MGMT gene promoter in glioblastomas. AB - In an effort to extend the potential relationship between the methylation status of MGMT promoter and response to CENU therapy, we examined the methylation status of MGMT promoter in 44 patients with glioblastomas. Tumor specimens were obtained during surgery before adjuvant treatment, frozen and stored at -80 degrees C until for DNA extraction process. DNA methylation patterns in the CpG island of the MGMT gene were determined in every tumor by methylation specific PCR (MSP). These results were then related to overall survival and response to alkylating agents using statistical analysis. Methylation of the MGMT promoter was detected in 68% of tumors, and 96.7% of methylated tumors exhibited also an unmethylated status. There was no relationship between the methylation status of the MGMT promoter and overall survival and response to alkylating agents. Our observations do not lead us to consider promoter methylation of MGMT gene as a prognostic factor of responsiveness to alkylating agents in glioblastomas. PMID- 15332331 TI - High and low grade oligodendrogliomas (ODG): correlation of amino-acid and glucose uptakes using PET and histological classifications. AB - Classification and treatment strategy of oligodendrogliomas (ODG) remain controversial. Imaging relies essentially on contrast enhancement using CT or MRI. The aim of our study was to use positron emission tomography (PET) using [18F]-flurodeoxyglucose (FDG) and [11C]-L-methyl-methionine (MET) to evaluate metabolic characteristics of ODG. We studied 19 patients with proven ODG, comparing standardized uptake values (SUV) and maximal tumor/contralateral normal tissues ratios (T/N). Imaging findings were compared with WHO, Smith and Daumas Duport classifications. Uptake of FDG was decreased only in 8 patients, independently of grading, while MET uptake was always increased. MET uptake was significantly higher for high grade tumors grouped according to Smith or Daumas Duport classifications, while no significant difference in MET uptake was found when using WHO classification. A different correlation was found between FDG and MET uptakes in normal tissues and high grade tumors. A trend for improved progression free survival was found for tumors that lacked contrast enhancement on MRI or those showing low FDG or MET uptake. In conclusion, MET appeared more sensitive than FDG to detect proliferation in ODG. The preferential protein metabolism, already noticeable for low-grade tumor, correlated with glucose metabolism and helped to separate, in vivo, high and low grade tumors. PMID- 15332333 TI - Radiotherapy for pediatric central nervous system tumors: a regional cancer centre experience. AB - INTRODUCTION: The purpose of this review was to analyze outcomes for pediatric patients treated for more common (non-low grade glioma) primary central nervous system (CNS) tumors at a Regional (tertiary) Cancer Center. Comparison to reported results from other regional centres and results from the contemporary literature were made. MATERIAL AND METHODS: The records of pediatric patients treated with radiotherapy at the London Regional Cancer Center (LRCC) for more common (non-low grade glioma) primary CNS tumors between 1980 and 2001 were reviewed. Details regarding tumor presentation, treatment and outcome were analyzed. RESULTS: Eighty-eight patients were eligible for the review. Twenty nine patients with malignant glioma, 37 patients with medulloblastoma or primitive neuroectodermal tumor (PNET), 15 patients with brainstem glioma, 4 with ependymoma and 3 with germ cell tumors were treated during this time period. Average follow-up for the group was 5 years (range 4 months to 19 years). Five year overall, progression free and cause specific survival were 45, 42 and 50%, respectively. For patients with malignant glioma median progression free and overall survival was 20 and 29 months. For patients with brainstem glioma median progression free and overall survival was 9 and 13 months. For medulloblastoma, 5 year progression free, and overall survival was 60 and 59%. CONCLUSIONS: RESULTS of this retrospective review of pediatric patients treated at a regional cancer center for primary CNS tumors (other than low grade glioma) were comparable to contemporary results reported by other Canadian centers and North American co operative group trials. PMID- 15332334 TI - Telling the story. Trustees and grassroots political advocacy. AB - As volunteers with a sophisticated understanding of hospitals, trustees provide a unique, vital link between the legislature, the hospital and the community. That's why grassroots political advocacy should be at the top of your board's "to do" list. PMID- 15332335 TI - Data in, safety out. Balanced scorecards help the board make patient safety their No. 1 priority. AB - Patient safety is a top board priority at The Nebraska Medical Center in Omaha. Trustees rely on data displayed in a balanced scorecard to help them lead the organization on its road to improved safety. PMID- 15332336 TI - Trustee workbook 3. Maximizing hospital revenue through retail. PMID- 15332337 TI - Avoiding scandal. Recommended practices for board executive compensation committees. AB - The executive compensation committee walks a fine line between providing competitive executive pay and risking regulatory scrutiny. Here's how you can keep your balance. PMID- 15332338 TI - The trustee's responsibility for quality care. PMID- 15332339 TI - Breaking the language barrier. Converting forms. PMID- 15332340 TI - Breaking the language barrier. Helping hands. PMID- 15332341 TI - Melanopsin--shedding light on the elusive circadian photopigment. AB - Circadian photoentrainment is the process by which the brain's internal clock becomes synchronized with the daily external cycle of light and dark. In mammals, this process is mediated exclusively by a novel class of retinal ganglion cells that send axonal projections to the suprachiasmatic nuclei (SCN), the region of the brain that houses the circadian pacemaker. In contrast to their counterparts that mediate image-forming vision, SCN-projecting RGCs are intrinsically sensitive to light, independent of synaptic input from rod and cone photoreceptors. The recent discovery of these photosensitive RGCs has challenged the long-standing dogma of retinal physiology that rod and cone photoreceptors are the only retinal cells that respond directly to light and has explained the perplexing finding that mice lacking rod and cone photoreceptors can still reliably entrain their circadian rhythms to light. These SCN-projecting RGCs selectively express melanopsin, a novel opsin-like protein that has been proposed as a likely candidate for the photopigment in these cells. Research in the past three years has revealed that disruption of the melanopsin gene impairs circadian photo- entrainment, as well as other nonvisual responses to light such as the pupillary light reflex. Until recently, however, there was no direct demonstration that melanopsin formed a functional photopigment capable of catalyzing G-protein activation in a light-dependent manner. Our laboratory has recently succeeded in expressing melanopsin in a heterologous tissue culture system and reconstituting a pigment with the 11-cis-retinal chromophore. In a reconstituted biochemical system, the reconstituted melanopsin was capable of activating transducin, the G-protein of rod photoreceptors, in a light-dependent manner. The absorbance spectrum of this heterologously expressed melanopsin, however, does not match that predicted by previous behavioral and electophysiological studies. Although melanopsin is clearly the leading candidate for the elusive photopigment of the circadian system, further research is needed to resolve the mystery posed by its absorbance spectrum and to fully elucidate its role in circadian photoentrainment. PMID- 15332342 TI - Circadian and short-term regulation of blood pressure and heart rate in transgenic mice with cardiac overexpression of the beta1-adrenoceptor. AB - Congestive heart failure is associated with a loss of circadian and short-term variability in blood pressure and heart rate. In order to assess the contribution of elevated cardiac sympathetic activity to the disturbed cardiovascular regulation, we monitored blood pressure and heart rate in mice with cardiac overexpression of the beta1-adrenoceptor prior to the development of overt heart failure. Telemetry transmitters for continuous monitoring of blood pressure and heart rate were implanted in 8 to 9-week-old wildtype and transgenic mice, derived from crosses of heterozygous transgenic (line beta1TG4) and wildtype mice. Cardiovascular circadian patterns were analyzed under baseline conditions and during treatment with propranolol (500mg/L in drinking water). Short-term variability was assessed by spectral analysis of beat-to-beat data sampled for 30min at four circadian times. Transgenic beta1TG4 mice showed an increase in 24h heart rate, while blood pressure was not different from wildtype controls. Circadian patterns in blood pressure and heart were preserved in beta1TG4 mice. Addition of propranolol to the animals' drinking water led to a reduction in heart rate and its 24 h variation in both strains of mice. Short-term variability in blood pressure was not different between wildtype and beta1TG4 mice, but heart rate variability in the transgenic animals showed a rightward shift of the high frequency component in the nocturnal activity period, suggesting an increase in respiratory frequency. In conclusion, the present study shows that both the circadian and the short-term regulation of blood pressure and heart rate are largely preserved in young, nonfailing beta1-transgenic mice. This finding suggests that the loss of blood pressure and heart rate variability observed in human congestive heart failure cannot be attributed solely to sympathetic overactivity but reflects the loss of adrenergic responsiveness to changes in the activity of the autonomic nervous system. PMID- 15332343 TI - Infradian rhythmicity in sleep/wake ratio in developing infants. AB - Although there are several reports on ultradian and circadian rhythms in newborns, we found only one report in which infradian periodicities are described for heart-rate measurements in the early stages of human development. Here, we report infradian rhythms in the monthly range in the sleep/wake cycle of four infants studied along 24 consecutive weeks. Our procedure was applied to sleep diary records from four healthy newborns. The data were arranged in binary time series representing sleep (-1) or wake (1) states. These time series were integrated in order to obtain the cumulative sleep/wake time. A measure of the sleep/wake ratio (SWR) was obtained by computing the average slope of the cumulative sleep/wake time. To extract periodicities we applied the Fourier periodogram to the temporal course of the SWR. We found a notorious difference in the SWR pattern among infants. In two infants the SWR showed a marked linear decay, spending more time asleep than awake, while in the two other infants oscillated near zero. We found robust oscillations in all children. In all cases the Fourier periodogram results present significant power in the infradian range. From these results, we suggest that sleep and wake durations are probably modulated by some internal stimuli. PMID- 15332344 TI - Genetic variability of arylalkylamine-N-acetyl-transferase (AA-NAT) gene and human sleep/wake pattern. AB - The cyclic production and secretion of melatonin has been associated with the sleep/wake cycle as well as other circadian rhythms. Since arylalkylamine-N acetyl-transferase (AA-NAT) is the rate-limiting enzyme responsible for the production of melatonin, it has been postulated to determine the circadian oscillations of melatonin. Genetic variability of the AA-NAT gene may therefore potentially influence sleep patterns in the normal population. In this study, a sleep pattern survey was performed in 210 students. Five subjects with early sleep onset time and long sleep duration (early/long sleepers), and 5 subjects with late sleep onset time and short sleep duration (late/short sleepers) were identified for genetic studies. All 10 subjects had identical sequences throughout the coding regions of the AA-NAT gene. In the promoter region, a 263G/C (relative to the transcription start site) single nucleotide polymorphism (SNP) was observed in 4 of the 5 late/short sleepers, but in only 1 of 5 early/long sleepers. The -263G/C SNP may therefore be an important determinant of the late/short sleep pattern. PMID- 15332345 TI - Sleep patterns of day-working, evening high-schooled adolescents of Sao Paulo, Brazil. AB - Children who grow up in developing countries of the world must work to help financially support their families, and they must also attend school. We investigated the impact of work on the sleep of working vs. nonworking high school students. Twenty-seven Sao Paulo, Brazil, public high school students (eight male and eight female working students plus six nonworking female and five nonworking male students) 14-18 yrs of age who attended school Monday-Friday between 19:00 to 22:30h participated. A comprehensive questionnaire about work and living conditions, health status, and diseases and their symptoms was also answered. The activity level and rest pattern (sleep at night and napping during the day) were continuously assessed by wrist actigraphy (Ambulatory Monitoring, USA). The main variables were analyzed by a two-factor ANOVA with application of the Tukey HSD test for multiple comparisons, and the length of sleep during weekdays vs. weekends was compared by Student t-test. Working students went to sleep earlier weekends [F(1,23)=6.1; p=0.02] and woke up earlier work days than nonworking students [F(1,23) = 17.3; p = 0.001]. The length of nighttime sleep during weekdays was shorter among all the working [F(1,23)= 16.7; p <0.001] than all the nonworking students. The sleep duration of boys was shorter than of girls during weekends [F(1,23)= 10.8; p <0.001]. During weekdays, the duration of napping by working and nonworking male students was shorter than nonworking female students. During weekdays working girls took the shortest naps [F(1,23)= 5.6; p = 0.03]. The most commonly reported sleep complaint during weekdays was difficulty waking up in the morning [F(1,23) = 6.5; p = 0.02]. During weekdays, the self-perceived sleep quality of working students was worse than nonworking students [F(1,23) = 6.2; p = 0.02]. The findings of this study show that work has negative effects on the sleep of adolescents, with the possible build-up of a chronic sleep debt with potential consequent impact on quality of life and school learning. PMID- 15332346 TI - Thermoregulation during mild exercise at different circadian times. AB - Eight healthy subjects exercised at 90watts on a cycle ergometer on four occasions, at times close to the minimum, maximum rate of rise, maximum, and maximum rate of fall of their resting core temperature. The duration of exercise was determined by the time taken for the core (rectal) temperature to reach an equilibrium value. Forearm skin blood flow and temperature were measured regularly during the exercise, as were heart rate and ratings of perceived exertion. Sweat loss was calculated by weighing the subjects nude before and after the exercise. The rise of heart rate was not significantly different at the four times of exercise, though the rating of perceived exertion was greatest at 05:00 h. Resting core temperatures showed a significant circadian rhythm at rest (the timing of which confirmed that exercise was being performed at the required times), but the amplitude of this rhythm was decreased significantly by the exercise. The initial rate of rise of core temperature, and the total rise from the resting to the equilibrium value, were both inversely proportional to resting temperature. The time-course of the rise was accurately described by a negative exponential model, but this model gave no evidence that the kinetics of the equilibration process depended upon the time of day. The thermoregulatory responses to the rise in core temperature--the amount of total sweat loss and rises in forearm skin blood flow and temperature--differed according to the time of exercise. In general, the responses were significantly greater at 17:00h compared with 05:00h, and at 23:00 h compared with 11:00 h. The results accord with predictions made on the basis of previous work by us in which core temperature rhythms have been separated into components due to the endogenous body clock and due to the direct effects of spontaneous activity. The results are discussed in terms of the ecological implications of the differing capabilities of humans to deal with heat loads produced by spontaneous activity or mild exercise at different phases of the circadian rhythm of resting core temperature. PMID- 15332347 TI - Administration-time-dependent effects of doxazosin GITS on ambulatory blood pressure of hypertensive subjects. AB - Previous studies have shown that a single nighttime dose of standard doxazosin, an alpha-adrenergic antagonist, reduces blood pressure (BP) throughout the 24 h. We investigated the administration-time-dependent effects of the new doxazosin gastrointestinal therapeutic system (GITS) formulation. We studied 91 subjects (49 men and 42 women), 56.7+/-11.2 (mean+/-SD) yrs of age with grade 1-2 essential hypertension; 39 patients had been previously untreated, and the remaining 52 had been treated with two antihypertensive medications with inadequate control of their hypertension. The subjects of the two groups, the monotherapy and polytherapy groups, respectively, were randomly assigned to receive the single daily dose of doxazosin GITS (4 mg/day) either upon awakening or at bedtime. BP was measured by ambulatory monitoring every 20 min during the day and every 30 min at night for 48 consecutive hours just before and after 3 months of treatment. After 3 months of doxazosin GITS therapy upon awakening, there was a small and nonstatistically significant reduction in BP (1.8 and 3.2mm Hg in the 24 h mean of systolic and diastolic BP in monotherapy; 2.2 and 1.9mm Hg in polytherapy), mainly because of absence of any effect on nocturnal BP. The 24 h mean BP reduction was larger and statistically significant (6.9 and 5.9 mm for systolic and diastolic BP, respectively, in monotherapy; 5.3 and 4.5 mm Hg in polytherapy) when doxazosin GITS was scheduled at bedtime. This BP-lowering effect was similar during both the day and nighttime hours. Doxazosin GITS ingested daily on awakening failed to provide full 24h therapeutic coverage. Bedtime dosing with doxazosin GITS, however, significantly reduced BP throughout the 24h both when used as a monotherapy and when used in combination with other antihypertensive pharmacotherapy. Knowledge of the chronopharmacology of doxazosin GITS is key to optimizing the efficiency of its BP-lowering effect, and this must be taken into consideration when prescribing this medication to patients. PMID- 15332348 TI - Possible evidence for shift work schedules in the media workers of the ant species Camponotus compressus. AB - The locomotor activity rhythm of the media workers of the ant species Camponotus compressus was monitored under constant conditions of the laboratory to understand the role of circadian clocks in social organization. The locomotor activity rhythm of most ants entrained to a 24h light/dark (12:12h; LD) cycle and free-ran under constant darkness (DD) with circadian periodicities. Under entrained conditions about 75% of media workers displayed nocturnal activity patterns, and the rest showed diurnal activity patterns. In free-running conditions these ants displayed three types of activity patterns (turn-around). The free-running period (tau) of the locomotor activity rhythm of some ants (10 out of 21) showed period lengthening, and those of a few (6 out of 21) showed period shortening, whereas the locomotor activity rhythm of the rest of the ants (5 out of 21) underwent large phase shifts. Interestingly, the pre-turn-around tau of those ants that showed nocturnal activity patterns during earlier LD entrainment was shorter than 24 h, which became greater than 24 h after 6-9 days of free-run in DD. On the other hand, the pre-turn-around tau of those ants, which exhibited diurnal patterns during earlier LD entrainment, was greater than 24 h, which became shorter than 24 h after 6-9 days of free-run in DD. The patterns of activity under LD cycles and the turn-around of activity patterns in DD regime suggest that these ants are shift workers in their respective colonies, and they probably use their circadian clocks for this purpose. Circadian plasticity thus appears to be a general strategy of the media workers of the ant species C. compressus to cope with the challenges arising due to their roles in the colony constantly exposed to a fluctuating environment. PMID- 15332349 TI - Phase-synchronization of daily motor activities can reveal differential circadian patterns. AB - The aim of the study was to determine any alteration of the 24 h motor activity pattern of a bipolar patient in different mood states. Actigraphic records were collected on an outpatient basis for a total of 387 days. The daily actograms were synchronized in phase to the time of morning awakening before averaging, which significantly enhanced the structure of the averaged traces. The actograms were divided into three groups based on total daily count. The daily motor activity patterns of the low- and high-activity days have a different circadian pattern. We propose it may have a relevance to the different mood states. The phase-synchronization of the 24h actograms to the patient's sleep-wake cycle, specifically to the time of awaking from the nighttime sleep, may help reveal differences in the daily temporal patterns of motor activity. PMID- 15332350 TI - Congenital anophthalmia: a circadian rhythm study. AB - A circadian rhythm of heart rate and respiratory rate was seen at 1, 8, and 12 months of age in an infant born without ocular tissue, which supports the possibility that the time cues were nonphotic. No melatonin circadian rhythm was detected at any age up to 9 years of age, and this is most likely associated with the anophthalmia and lack of photic input to the suprachiasmatic nucleus. Usually circadian organization is present after the neonatal period and approaches adult levels with development. PMID- 15332351 TI - Nuclear and isotopic techniques application used in supporting nutritional studies in Latin America countries. AB - The global nutrition community recognizes the usefulness of nuclear and isotopic techniques and especially stable isotopes for accurate measurements in key areas of human nutrition and health. Although progress has been made in many Latin America countries in reducing the absolute number of undernourished people, food policy and public health communities have to face the double burden on health. On one side they have to address the prevention of obesity and on the other side they are trying to reduce undernutrition. However, to be successful in development of sustainable nutrition programs, a combination of relevant scientific knowledge and approaches that are practical is required to assure even a reasonable level of success. The objectives of this review are to highlight the role of isotopic techniques in nutritional studies and to review the role of the International Atomic Energy Agency in supporting nutrition interventions in Latin America countries. PMID- 15332352 TI - [Pica during pregnancy: a frequently underestimated problem]. AB - Pica is the compulsive intake of non-nutritive substances such as earth, clay, chalk, soap and ice. The most common forms of pica are geophagia or the intake of earth and pagophagia or the intake of ice. The description of this peculiar phenomenon dates back to the Greco-Roman civilization. Its prevalence during pregnancy is generally underestimated. Published data reveal a prevalence of between 8% and 65%. Investigations from Latin America indicate a prevalence of 23% to 44%. It is not clear yet which are the causes that predispose to pica, but they are frequently associated with anemia or iron deficiency during pregnancy. Its diagnosis, which only consists in questioning pregnant women, is generally omitted during prenatal care, probably because health professionals have no knowledge about this disorder. The identification of pica in pregnant women could contribute to the detection of a risk group where it is necessary to implement strategies as regards both the evaluation and the nutritional education. PMID- 15332353 TI - [Lactation and alcohol: clinical and nutritional effects]. AB - Researchers have turned their attention to the effects of alcohol consumption on breastfeeding, with significant negative findings concerning both the mother and the newborn. This study is a meta-analysis of the principle research performed in the last decade that was concerned with lactation and alcohol. Results from experimental and human subject research has shown that effects of alcohol include: behavioural changes, reduced milk and luteinising hormone production, with increased fat content, reduced lactose content. Increased lipogenesis and increased activity of lipoprotein and hormone-sensitive lipase, structural alterations in the epithelial cells of the breast and abnormal casein production, reduced oxytocin and prolactin production with subsequent reduced milk ejection, and reduced weight and protein content of the breast. Reduction in food consumption, body weight, growth and development and hepatic glycogen, serum glucose, amino-acids, insulin, glycerol, fatty acids and urea, and an increase in serum acetoacetic acid is seen in newborn children that were breastfed by animals with a high intake of alcohol during pregnancy or the puerperal period. Alcohol consumption during lactation caused a reduction in liver weight and triglyceride, protein, DNA and lipid content, in the newborns. Serum changes included a reduction in protein, triglyceride, cholesterol, fatty acid and glycerol level with an increase in beta-hydroxybutirate levels. Changes also included alterations in the motor system and behaviour. Further studies are needed to determine, with confidence, the minimum level of alcohol consumption that can provoke pathological effects in both the mother and the child. PMID- 15332354 TI - [Role of black bean Phaseolus vulgaris on the nutritional status of Guatemalan population]. AB - Guatemala provides an example of epidemiological superposition, in which health problems typical of developed countries and developing countries are both observed. Nutritional deficiencies in some micronutrients like vitamin A and iron coexist alongside chronic diseases such as diabetes type II and cardiovascular diseases. The importance of black beans in the normal Guatemala diet is well known:70g per capita of black beans are consumed daily. Black beans are an important sources of protein and energy in the diet. They contain "lente" digestion carbohydrates and a high proportion of non-digested carbohydrates that may be fermented in the large intestine. Theses types of carbohydrates are associated with a low glycemic response, low serum cholesterol levels, and a decrease of colon cancer risk factors. These physiological effects may be related to colonic fermentation end products (propionic and butyric acids). Black beans also contain several antinutritional compounds (enzymatic inhibitors, haemaglutenins, saponins and phytic acid, etc.), some of them thermolabiles that are partially eliminated during culinary processes and may modify the nutritional quality of beans. Black beans play a crucial role in the etiology of several diseases in Guatemala. PMID- 15332355 TI - Prematurity and maternal folate deficiency: anemia during pregnancy study group results in Valencia, Venezuela. AB - The purpose of this study was to determine the association and its magnitude between prematurity and folate deficiency in women in their third trimester of pregnancy, and at labor. An incident case-control study was conducted using 2 controls per case. Data was obtained in a tertiary hospital in Valencia, Venezuela. A total of 543 women who delivered between May and December 1996 entered into the study. Women having a preterm delivery (< 37 weeks of gestation at delivery) were defined as cases (n = 181). Anemia was defined according to WHO as Hb less than 11 g/dL, when a pregnant woman had a folate serum level < 3 microg/ml was considered a folate deficiency. Logistic regression was used to analyze the data and likelihood ratio test was done for model comparison. Folate deficiency was found to be significantly associated with prematurity (Odds Ratio: 1.97; 95%CI = 1.06 to 3.68 P = .032), after adjusting for prior preterm labor, prenatal care visits, prior abortion, prior fetal death, placental abruption, and premature rupture oval membranes. In conclusion, maternal folate deficiency at the end of the third trimester of pregnancy, at labor, was associated with an increased risk of prematurity. PMID- 15332356 TI - [An epidemiological index to assess the nutritional status of children based in a polynomial model of values from Z punctuation for the age in Mexico]. AB - A nutritional status index was built by modeling the mathematical function of the mean Z scores of weight for age, from 60,079 children under five years of age, selected in a probabilistic fashion from the Mexican population. The most precise mathematical model was a fifth degree polynomial. The correlation coefficient was between .937 0.001). CONCLUSIONS: There are size and location differences between non neoplastic polyps and adenomas. Adenomas associated to cancer may measure less than 5 mm. Therefore the polyp size criteria to decide surgical behavior in symptomatic gallstone patients may be misleading. PMID- 15332369 TI - [Dual X-ray absorptiometry [DEXA] in normal Chilean women. Correlation between reference values and anthropometric variables]. AB - BACKGROUND: Bone mineral density results are expressed as the number of standard deviations from reference values in similar populations. However these reference values have not been obtained locally and there may be discrepancies between Chilean and other Hispanic populations. AIM: To evaluate the results of DEXA in healthy Chilean women and to correlate them with reference values and anthropometric parameters. MATERIAL AND METHOD: We studied prospectively 166 normal women aged between 15 and 55 years, using a Hologic QDR 1000 equipment, yielding bone mineral density (BMD), bone mineral content (BMC) and bone area (A) in the lumbar spine (LS) and femoral neck (FN). These bone densitometric variables vere correlated with weight, height and age of women and compared against the reference values of the Hologic equipment. Relation between BMC and A was evaluated using a regression analysis. RESULTS: BMD and BMC were lower to reference values in most age groups, especially for LS. There was a significant correlation of BMD with A in LS (r: 0.278, p < 0.001), weght in both LS (r: 0.382, p < 0.0001) and FN (r: 0.266, p < 0.001), height in LS (r: 0.258, p < 0.001) and age in FN (r: -0.231, p < 0.01). Only in LS, regression analysis between A and BMC did not show direct proportionality. CONCLUSIONS: BMD of healthy Chilean women was lower than the reference values of the Hologic equipment for most age groups. Bone densitometric variables are non uniformly influenced by weight, height and size of bone structures (A) in normal women, especially in LS. Correct interpretation of DEXA requires the validation of reference values and to assess the value of bone area measurements. PMID- 15332370 TI - [Carcinoembryonic antigen (CEA) as an independent prognostic factor in colorectal carcinoma]. AB - BACKGROUND: CEA is widely used in the follow up of patients with colorectal carcinoma. AIM: To study the value of preoperative CEA as an independent prognostic factor in colorectal carcinoma. PATIENTS AND METHODS: Analysis of 373 operated patients (204 females, age range 21-92 years) with colorectal carcinoma and a mean follow up of 53 months. The cutoff value for CEA was 5 ng/ml. Ninety four percent of patients had an excisable tumor, 79% had involvement of perirectal/pericolonic adipose tissue and 46% had lymph node involvement. Staging was done using Dukes-Turnbull and TNM classifications. RESULTS: CEA was normal in 61% of cases, over 5 ng/ml in 39% and over 15 ng/ml in 22%. There was a strong correlation between mean preoperative CEA and tumor stage, depth and lymph node involvement. During the follow up, 140 patients died, 57 with normal and 83 with elevated CEA. Cancer mortality in patients subjected to a curative excision of the tumor (Dukes A-C2/TNM I-III) was 9% for colonic tumors and 36% for rectal tumors (p < 0.001). There were no survival differences in patients with Dukes B/TNM II tumors according to preoperative CEA. Among Dukes C/TNM III tumors, survival difference was only significant for rectal tumors. A Cox model disclosed tumor stage, location and preoperative CEA as independent prognostic factors for survival. CONCLUSIONS: CEA is an independent prognostic factor for survival in colorectal carcinoma and high levels suggest an advanced disease. PMID- 15332371 TI - [Assessment of 1497 Chilean nursing home residents, using the Resource Utilization Group method, RUG T-18]. AB - BACKGROUND: There is little information about Chilean elderly residents of long term care facilities, regarding their characteristics and need for resources. AIM: To describe main characteristics and resource utilization of residents of one of the largest nursing homes in Chile, Fundacion Las Rosas de Ayuda Fraterna. MATERIAL AND METHODS: In a cross sectional and descriptive study, all residents were evaluated using the RUG T-18 method, that assess activities of daily living and the complexity of their clinical situation. RESULTS: We assessed 1497 subjects 60 years old and over (73% women), with an age range of 60-106 years. Thirty six percent had urinary incontinence, 19% required assistance for feeding, and 38% needed help for walking or moving. Fifty seven percent were in the lowest category of complexity, "Institutionalization". Very few residents were in the most demanding categories, no one classified as "Rehabilitation", and only 0.7% were in "Special Care". CONCLUSIONS: This study is an important start point to learn more about elderly subjects living in nursing homes in Chile. PMID- 15332373 TI - [Change in reproductive partner among women delivering at a Metropolitan Santiago public hospital]. AB - BACKGROUND: Previous studies in Metropolitan Santiago, showed that at least 10% of mothers changed their reproductive partner. AIM: To study the frequency of reproductive partner change and its associated factors among women delivering in a public hospital. SUBJECTS AND METHODS: We interviewed 859 non primiparous women that gave birth to a child in a public hospital of Metropolitan Santiago, between March 2001 and February 2002. RESULTS: Mean maternal age rose according to birth order from 27.4 to 33.6 years from the second to the fourth sibling. Married women were the largest group and single ones decreased from the second to the third sibling. Twenty two percent had some activity out of home and those with a high birth order (fourth or more) exhibited poorer schooling. Unwanted pregnancies increased with birth order, reaching 55% among "fourth and over" births. Thirty four percent of women delivering their second child, changed their partner. Among women delivering their third child, 38.6% had one sibling with a different father; 20.4% had changed partner during their second pregnancy and 20% during the current one. Among those in the "fourth and over, group, 56.1% had at least one sibling with a different father; 18.3% had changed partner in the second sibling; 15%, in the third and 27.8%, in the fourth and over. In all birth orders, mothers with activity out of home exhibited a higher frequency of partner change. The most common reasons to change partner were infidelity, abandon or a personal decision, while economic reasons were uncommon. CONCLUSIONS: A great number of children are exposed to bonding ruptures and to the consequential mental health or behavioral problems. PMID- 15332372 TI - [Relationship between the cause of admission and the cause of death in patients admitted to an internal medicine departament]. AB - BACKGROUND: When patients are admitted to an internal medicine ward, they and their relatives believe that they will leave the internal medicine service in a better health condition or, if they die, their cause of death will be the same that motivated the hospital admission. Sometimes, patients die due to a different disease than the cause of admission or due to complications of diagnostic or therapeutic procedures. AIM: To assess the proportion of deaths that occurred in internal medicine wards for causes that differ from the cause for admission and the proportion of deaths related to complications of diagnostic or therapeutic procedures. PATIENTS AND METHODS: Three hundred and three consecutive patients were studied during 28 months. Their cause qf death was classified n four groups: A. If it was the same illness of admission; B. Afrecuent or expected complication of the illness that motivated the admission; C. A complication of one of their chronic diseases and not the one of admission; D. If it was not related to an already present chronic illness or the cause for admission. RESULTS: Sixty nine percent of deaths corresponded to group A, 7.9% to group B, 11.2%96 to group C and 11.9% to group D. Diagnostic or therapeutic procedures were the cause of death in 0. 7 and 3.6% of cases respectively. CONCLUSIONS: Twenty three percent of patients died due to causes that differ from the cause of admission. A low percentage of deaths were related to procedures and most of them were due to complications of therapeutic measures. PMID- 15332374 TI - [Liaison psychiatry in two public hospitals of Santiago]. AB - BACKGROUND: Medical and surgical teams consult psychiatrists when abnormal behavior or psychopathological traits are observed in their patients. AIM: To describe the experience of liaison psychiatry teams in public hospitals. MATERIAL AND METHODS: Analysis of 128 consultations, involving 110 patients. Eighty eight patients were subjected to the structured Clinical Interview for DSM-IV-TR Axis I Disorders--Patient Edition, the Mini Mental State score and to a search for personality disorders. RESULTS: The most commonly consulted problems were depression in 44 patients, substance abuse in 29 and suicidal behavior in 21. The psychiatric study detected that 8% of patients were asymptomatic, 8% had stress reactions, 16.5% had substance abuse, 14.6% had alcohol abuse, 11% had a major depression, 10% had delirium or dementia, 9% had anxiety disorders, 4% schizophrenia, 18% personality disorders and 19% had suicidal behaviors. CONCLUSIONS: Psychiatric consultations in medical and surgical services are common and have a wide variety of severity. PMID- 15332375 TI - [Congenital hepatic fibrosis. Report of five cases]. AB - BACKGROUND: Congenital hepatic fibrosis (CHF) is an autosomic dominant disease that has been associated with polycystic kidney disease. AIM: To describe the medical management of 5 children with CHF and to evaluate the presence and extension of the associated renal disease. PATIENTS AND METHODS: Retrospective review of the medical charts of 5 children with CHF, aged 2 to 14 years. RESULTS: Three children presented autosomic recessive polycystic kidney disease, which was diagnosed before the appearance of liver disease manifestations. They presented a more severe liver damage, with a more aggressive clinical course requiring use of transjugular intrahepatic porto-systemic shunts (TIPS) or surgical porto-systemic shunts to control portal hypertension. The other two children, in whom the diagnosed was based on asymptomatic hepatomegaly, had normal renal function and structure with a more benign clinical course. CONCLUSIONS: The diagnosis of CHF should be suspected not only in children with polycystic kidney disease but in those children with persistent, hard consistency, left lobe predominance hepatomegaly. PMID- 15332376 TI - [Acute interstitial nephritis associated to lamotrigine use. Report of one case]. AB - Acute interstitial nephritis is a mononuclear and sterile inflammation of the renal interstice caused by drugs, infections or immune phenomena. The clinical presentation is characterized by the triad of rash, fever and eosinophilia. We report a 32 years old man, in treatment with lamotrigine for depression, admitted to the hospital due to fever, abdominal pain, jaundice, cutaneus rash and malaise. Due to an oliguric renal failure of acute onset, a renal biopsy was done. The pathological study showed a granulomatous acute interstitial nephritis. He was started on hemodialysis and treated with cessation of the drug and corticosteroids, with complete recovery of the renal function. PMID- 15332377 TI - [Peri-ileostomic pyoderma gangrenosum. Report of one case]. AB - We report a 39 years old woman with Ulcerative Colitis unresponsive to medical treatment. A total colectomy with Brooke ileostomy was performed, as the first operation. After six weeks she developed a peri-ileostomic Pyoderma Gangrenosum. She was treated by a multidisciplinary team, and after seven months with local treatment and systemic medication, the skin healed. Her bowel transit was reconstricted later with an ileoanal pouch and she was reinserted into her previous job. PMID- 15332378 TI - [Analysis of the new health module of the National Socioeconomic Characterization Survey 2000]. AB - BACKGROUND: The National Socioeconomic Characterization Survey (CASEN) was modified in the year 2000, to include an assessment of the inequalities in the access to health services. AIM: To analyze the health issues of year 2000 survey database. MATERIAL AND METHODS: During 2000, 38338 urban and 26698 rural dwellings were surveyed, totaling 240000 people analyzing ascription to public or private health services and the need demand and use of these services. RESULTS: A higher risk population (lower income, higher age and women) is ascribed to public health services. Sixty five percent of the population self perceives their health as good, this figure decreases along with age and women have a worse self perception than men. In the 30 days prior to the survey, 13% of the population had a health related event; this figure was higher among women and the lower income quintiles. Expressed health demand was higher among women and lower income quintiles. A multivariate analysis identified an age below 14 years, pertaining to a minority ethnic group, ascription to private health services, residing in rural areas, pertaining to the lower income quintile and male sex, as factors associated to a lack access to health care. In the adjusted model, pertaining to the National Health Fund (a public system) is a protective factor to receive health services. CONCLUSIONS: The significant inequalities in the access to health care should be corrected with the new Health Reform. The new module incorporated to the National Socioeconomic Characterization Survey, is useful to assess the access to health care in Chile. PMID- 15332379 TI - [Atmospheric pollution and cardiovascular damage]. AB - The damaging effect of atmospheric pollution with particulate matter and toxic gases on the respiratory system and its effect in the incidence and severity of respiratory diseases, is well known. A similar effect on the cardiovascular system is currently under investigation. Epidemiological studies have demonstrated that the inhalation of particulate matter can increase cardiovascular disease incidence and mortality, specially ischemic heart disease. The damage would be mediated by alterations in the autonomic nervous system, inflammation, infections and free radicals. In human studies, environmental pollution is associated with alterations in cardiac frequency variability and blood pressure and with changes in ventricular repolarization. Experimentally, an enhancement of ischemia, due to coronary obstruction, has been demonstrated. The study of the toxic effects of environmental pollution over the cardiovascular system, is an open field, specially in Chile, were the big cities have serious contamination problems. PMID- 15332380 TI - [The change in medical practice. Psycho-social challenges for the profession]. AB - This article analyzes important changes in medical practice, focusing in those most deeply perceived by a group of physiciansfrom the Metropolitan Region of Santiago, included in a joint research conducted by the Departments of Psychiatry and Mental Health (East) of the Medical Faculty at the University of Chile and Public Health of the Catholic University of Santiago, during 2003. These are the perceived changes in the relationship between doctors and patients; increased limits in professional autonomy and the fragmentation of medical practice. Reflecting transformations in social relationships in general, they have added new stress and frustrations, as well as new opportunities and rewards to the medical profession. The perceptions identified are commented within the structural determinants of medical practice. These issues call for a refreshing discussion on the values supporting medical professionalism and the concept of profession in itself, in view of the challenges posed by the current social and cultural changes. PMID- 15332381 TI - [Dangerous spiders in Chile]. PMID- 15332382 TI - Marching to the beat of different drummers: individual airway response diversity. PMID- 15332383 TI - Stroke alert: sleep disordered breathing predicts survival? PMID- 15332384 TI - Blunted sensation of dyspnoea and near fatal asthma. PMID- 15332385 TI - Patient-physician communication about end-of-life care for patients with severe COPD. AB - Since patients with chronic obstructive pulmonary disease (COPD) infrequently discuss treatment preferences about end-of-life care with physicians, the goal of the present study was to identify which specific areas of communication about end of-life care occur between patients with severe COPD and their physicians, and how patients rate the quality of this communication. A total of 115 patients with oxygen-dependent COPD, identified in pulmonary clinics in three hospitals and through an oxygen delivery company, were enrolled in this study. A 17-item quality of communication questionnaire (QOC) was administered to patients, along with other measures, including satisfaction with care. The patients reported that most physicians do not discuss how long the patients have to live, what dying might be like or patients' spirituality. Patients rated physicians highly at listening and answering questions. Areas patients rated relatively low included discussing prognosis, what dying might be like and spirituality/religion. Patients' assessments of physicians' overall communication and communication about treatment correlated well with the QOC. Patients' overall satisfaction with care also correlated significantly with the QOC. In conclusion, this study identifies areas of communication that physicians do not address and areas that patients rate poorly, including talking about prognosis, dying and spirituality. These areas may provide targets for interventions to improve communication about end-of-life care for patients with chronic obstructive pulmonary disease. Future studies should determine the responsiveness of these items to interventions, and the effect such interventions have on patient satisfaction and quality of care. PMID- 15332386 TI - The TORCH (towards a revolution in COPD health) survival study protocol. AB - Only long-term home oxygen therapy has been shown in randomised controlled trials to increase survival in chronic obstructive pulmonary disease (COPD). There have been no trials assessing the effect of inhaled corticosteroids and long-acting bronchodilators, alone or in combination, on mortality in patients with COPD, despite their known benefit in reducing symptoms and exacerbations. The "TOwards a Revolution in COPD Health" (TORCH) survival study is aiming to determine the impact of salmeterol/fluticasone propionate (SFC) combination and the individual components on the survival of COPD patients. TORCH is a multicentre, randomised, double-blind, parallel-group, placebo-controlled study. Approximately 6,200 patients with moderate-to-severe COPD were randomly assigned to b.i.d. treatment with either SFC (50/500 microg), fluticasone propionate (500 microg), salmeterol (50 microg) or placebo for 3 yrs. The primary end-point is all-cause mortality; secondary end-points are COPD morbidity relating to rate of exacerbations and health status, using the St George's Respiratory Questionnaire. Other end-points include other mortality and exacerbation end-points, requirement for long-term oxygen therapy, and clinic lung function. Safety end-points include adverse events, with additional information on bone fractures. The first patient was recruited in September 2000 and results should be available in 2006. This paper describes the "TOwards a Revolution in COPD Health" study and explains the rationale behind it. PMID- 15332387 TI - Heterogeneity of narrowing in normal and asthmatic airways measured by HRCT. AB - Asthmatic airway narrowing is heterogeneous and contributes to airway hyperresponsiveness. The present study compared heterogeneity of narrowing during methacholine challenge in asthmatics and normal subjects using high-resolution computed tomography (HRCT). The current authors defined heterogeneity as variability in narrowing greater than the repeatability of measurement. Airways of <2 mm diameter were compared with larger airways from baseline and postmethacholine HRCT of the right lower lung in 13 normals (seven had repeat baseline scans) and seven asthmatics. The coefficient of repeatability was calculated from repeat scans (RepAi) and was compared with heterogeneity of narrowing measured by the variability in narrowing from pre versus postmethacholine scans (VardeltaAi). Forced expiratory volume in one second decreased 27+/-6% and 24+/-8% in normals and asthmatics, respectively. Airways >2 mm narrowed more heterogeneously in asthmatics (VardeltaAi=+/-0.85 mm) compared with normals (VardeltaAi=+/-0.67 mm), with both being greater than the measure of repeatability (RepAi=+/-0.16 mm). Small airway narrowing was not heterogeneous in asthmatics (VardeltaAi=+/-0.59 mm) or normals (VardeltaAi=+/-0.53 mm) compared with repeatability (RepAi=0.51 mm). It is possible to study heterogeneity of airway narrowing in small and large airways using high resolution computed tomography. Airway narrowing is heterogeneous in the large airways of asthmatics and normals, being greater in asthmatics. PMID- 15332388 TI - Dyspnoea at rest and at the end of different exercises in patients with near fatal asthma. AB - Blunted perception of dyspnoea under resistive loading has been observed in patients with a history of near-fatal asthma (NFA). The perception of dyspnoea at rest and at the end point of various exercises was assessed in such patients. Respiratory function and exercise capacity (6-min walking distance, incremental cycloergometry and inspiratory threshold loading) were assessed in seven NFA and eight non-NFA patients. Dyspnoea (Borg scale) was measured at rest and at the end point of the various exercises. Dyspnoea at rest was significantly lower in NFA patients. Although exercise tolerance was similarly reduced in both the NFA and non-NFA groups, dyspnoea at peak cycle exercise was significantly lower in the former (2.6+/-2 versus 6.1+/-3.8 (Borg scale; mean+/-SD)), who mainly (86%) stopped because of leg discomfort. A similar trend was observed in the 6-min walking distance and inspiratory threshold loading tests. Dyspnoea at peak exercise was the best indicator of the NFA condition, with a sensitivity of 100% and specificity of 63% for a Borg scale score of < or = 6. Perception of dyspnoea is blunted in near-fatal asthma patients at both rest and the end point of various forms of exercise. Dyspnoea at peak exercise is the best indicator of the near-fatal asthma condition. PMID- 15332389 TI - Montelukast protects against nasal lysine-aspirin challenge in patients with aspirin-induced asthma. AB - Aspirin-induced asthma (AIA) is associated with increased production of cysteinyl leukotrienes (CysLT). Although leukotriene CysLT1-receptor antagonists improve lower airway outcomes in AIA, their effects and dose-response in the upper airway is less well documented. The present study evaluated the dose-response for montelukast (ML) against nasal lysine-aspirin challenge in patients with AIA. A total of 12 patients with a clear-cut history of AIA were randomised in double blind cross-over fashion to receive single doses of ML 10 mg, ML 40 mg, or placebo (PL), with nasal lysine-aspirin challenge performed 12 h after dosing. Measurements of peak nasal inspiratory flow (PNIF), nasal blockage visual analogue scale (VAS) and forced expiratory volume in one second (FEV1) were made over 120 min after nasal lysine-aspirin challenge. Prechallenge values for mean+/ SEM PNIF (L x min(-1)) were not significantly different comparing all groups: ML 10 mg (132+/-10), ML 40 mg (125+/-12) and PL (132+/-11). There was no significant difference comparing the maximum % PNIF fall from baseline between screening (46+/-6) and PL (45+/-6). The maximum % PNIF fall from baseline was significantly greater with PL (45+/-6) compared to either ML 10 mg (34+/-6) or ML 40 mg (32+/ 5). There was also a significantly greater mean % PNIF response over 120 min after lysine-aspirin challenge for PL (26+/-7) compared to either ML 10 mg (14+/ 6) or ML 40 mg (17+/-6). There were no significant differences for the maximum or mean % PNIF fall from baseline comparing ML 10 mg and ML 40 mg. A significant increase in nasal blockage VAS score was observed between baseline and 60 min or 120 min with PL but not with ML 10 mg or ML 40 mg. There were no significant differences for either the maximum or mean % FEV1 over 120 min as change from baseline comparing all groups. A single 10 mg dose of montelukast partially protected against the local effects of nasal lysine-aspirin challenge, with no further benefit at 40 mg. Nasal lysine-aspirin challenge appeared to be a reproducible and safe method in assessing patients with aspirin-induced asthma. PMID- 15332390 TI - Beta-adrenoceptor stimulation and neutrophil accumulation in mouse airways. AB - This study characterised the effect of beta-adrenoceptor stimulation on endotoxin induced accumulation of neutrophilic granulocytes in mouse airways, where the cytokines interleukin (IL)-6 and macrophage inflammatory protein (MIP)-2 are involed as mediators. The beta2-adrenoceptor agonist salbutamol (0.025-250 fMol) was administered intranasally in mice 24 h prior to administration of endotoxin (10 microg) intranasally. Bronchoalveolar lavage (BAL) fluid and venous blood, respectively, was harvested (6 or 24 h) after administration of endotoxin. Salbutamol substantially decreased the number of neutrophils in BAL fluid from endotoxin-exposed (6 and 24 h) mice and this effect was dose dependent (24 h). Pretreatment with the beta-adrenoceptor antagonist propranolol attenuated the inhibitory effect of salbutamol on BAL neutrophils (6 and 24 h), an attenuation that was not due to any unspecific effect of propranolol. Salbutamol also substantially decreased IL-6, but not MIP-2 in BAL fluid (6 h). In contrast to BAL fluid, salbutamol caused a moderate increase in blood neutrophils (24 h). In conclusion, as indicated in mouse airways in vivo, beta-adrenoceptor stimulation prior to endotoxin exposure inhibits the induced accumulation of neutrophils at a time point much later than that anticipated from its bronchodilatory effect. Even though the detailed molecular mechanisms behind this sustained "anti inflammatory" effect remain unknown, it seems likely that this effect is in part due to a decrease in the local concentration of interleukin-6. PMID- 15332391 TI - Reduced inspiratory flow attenuates IL-8 release and MAPK activation of lung overstretch. AB - Lung overstretch involves mechanical factors, including large tidal volumes (VT), which induce inflammatory responses. The current authors hypothesised that inspiratory flow contributes to ventilator-induced inflammation. Buffer-perfused rabbit lungs were ventilated for 2 h with 21%, O2+5%, CO2, positive end expiratory pressure of 2-3 cmH2O and randomly assigned to either: 1) normal VT (6 mL x kg(-1)) at respiratory rate (RR) 30, inspiration:expiration time ratio (I:E) 1:1, low inspiratory flow 6 mL x kg(-1) x s(-1); 2) large VT (12 mL x kg(-1)) at RR 30, I:E 1:1, high inspiratory flow 12 mL x kg(-1) x s(-1) (HRHF); 3) large VT at RR 15, I:E 1:1, low inspiratory flow 6 mL x kg(-1) x s(-1) (LRLF); or 4) large VT at RR 15, I:E 1:2.3, high inspiratory flow 10 mL x kg(-1) x s(-1) (LRHF). Physiological parameters, tumour necrosis factor (TNF)-alpha, interleukin (IL)-8 and activation of mitogen-activated protein kinases (extracellular signal regulated kinase (ERK)1/2, p38 and stress-activated protein kinase (SAPK)/ c-Jun N-terminal kinase (JNK)) were measured. HRHF increased weight gain, perfusate IL 8 and phosphorylation of ERK1/2, p38 and SAPK/JNK. These responses were absent during LRLF but present during LRHF. Changes in TNF-alpha were small. Tissue IL-8 and phospho-ERK1/2 staining was localised primarily to smooth muscle, adventitia and bronchial epithelium within larger bronchioles and arterioles. These results indicate that mild overstretch of perfused lungs during high inspiratory flow enhances inflammatory signalling by cells in lung regions most affected by strong turbulent airflow. PMID- 15332392 TI - Triggering receptor expressed on myeloid cells: role in the diagnosis of lung infections. AB - The triggering receptor expressed on myeloid cells (TREM)-1 is a recently described molecule, which plays an important role in myeloid cell-activated inflammatory responses. TREM-1 is expressed on blood neutrophils and monocytes, and also on alveolar macrophages, thus suggesting a potential role in lung inflammatory responses against infections. To investigate the differential expression of TREM-1 in lung infections, its levels were assessed in bronchoalveolar lavage specimens from patients with community-acquired pneumonia or tuberculosis. TREM-1 was also investigated in patients with interstitial lung diseases, as a model of noninfectious inflammatory disease of the lung. TREM-1 expression was significantly increased in lung neutrophils and in lung macrophages of patients with pneumonia (n=7; 387.9+/-61.4 and 660.5+/-18.3, respectively) compared with patients with pulmonary tuberculosis (n=7; 59.2+/ 13.1 and 80.6+/-291.2) and patients with interstitial lung diseases (n=10; 91.8+/ 23.3 and 123.9+/-22.8). In contrast, TREM-1 expression on peripheral blood neutrophils was no different among the three groups. In conclusion, these data suggest that triggering receptor expressed on myeloid cells-1 is selectively expressed in the lungs of patients with pneumonia caused by extracellular bacteria and not in patients with tuberculosis, providing a potential marker for differential diagnosis. PMID- 15332393 TI - Sarcoidosis and granuloma genes: a family-based study in African-Americans. AB - The evidence for a genetic component in the aetiology of sarcoidosis includes familial aggregation, associations with genetic polymorphisms, and linkage to the major histocompatibility complex class region on chromosome 6p. Unfortunately, the majority of genetic associations with sarcoidosis have not been consistently replicated. In the present study, using a family-based study design, which controls for population stratification, the authors attempted to replicate previously reported associations between sarcoidosis and three attractive candidate genes studied primarily in case-control samples. In 225 nuclear families, ascertained through African Americans with a history of sarcoidosis, no evidence was found for an association between sarcoidosis susceptibility and polymorphisms in the angiotensin converting enzyme, vitamin D receptor and tumour necrosis factor-alpha genes. Further analyses of chronic and acute disease phenotypes failed to reveal any notable associations. Assuming an underlying inheritance model with an additive allelic effect on disease risk, the current study had approximately 80-90% statistical power to detect a 3-fold increased risk associated with the putative risk allele of the polymorphisms under study. The present authors conclude that in African-Americans, the angiotensin converting enzyme, vitamin D receptor, and tumour necrosis factor-alpha genes are not significant risk factors for sarcoidosis susceptibility. PMID- 15332394 TI - The use of SPECT in preoperative assessment of patients with lung cancer. AB - Perfusion scintigraphy is the most frequently used method for the regional assessment of pulmonary function in candidates for pulmonary resection with borderline respiratory function. This method provides two-dimensional images, and it considers all the segments of the pulmonary lobes as having the same volume and function, without considering the spatial overlapping of pulmonary areas with different function. As single-photon emission computed tomography (SPECT) provides tomographic imaging, this could be a more precise method for regional assessment. In this study, the postoperative predicted forced expiratory volume in one second (FEV1) (FEV1,ppo) was calculated in 26 patients with lung cancer using FEV1, quantitative lung perfusion scan with planar acquisition (PA) and quantitative lung perfusion scan with tomographic imaging (SPECT). The estimated FEV1,ppo values obtained using both methods were compared with FEV1 values measured after surgery (mean: 48+/-44 days; range: 15-180 days; median: 32 days). The Pearson's linear correlation coefficient was 0.8840 for FEV1,ppo estimated by PA and 0.8791 for FEV1,ppo estimated by SPECT. The linear correlation coefficient for lobectomy was greater than the coefficient for pneumonectomy using both methods. In conclusion, both methods show good correlation for real postoperative pulmonary function without demonstrating single-photon emission computed tomography superiority over planar acquisition, and both methods were more effective for estimating postoperative predicted forced expiratory volume in one second in lobectomies than in pneumonectomies. PMID- 15332395 TI - Efficacious pleurodesis with OK-432 and doxorubicin against malignant pleural effusions. AB - Malignant pleural effusion develops frequently in patients with advanced lung cancer. Chemical pleurodesis is the most effective palliative treatment for these patients. The efficacy of pleurodesis using both OK-432, a preparation of Streptococcus pyogenes, and doxorubicin for 20 patients with cytology-proven malignant pleural effusion associated with lung cancer was evaluated. After complete removal of pleural effusion, OK-432 and 30 mg of doxorubicin were injected via an inserted chest tube. Treatment was terminated when the volume of daily drainage reached <200 mL. If the daily volume remained >200 mL, an additional OK-432 was administered every 3 days. In total, 16 patients (80%) revealed a complete response, two patients (10%) revealed a partial response, and no response was seen in two patients. Eighteen patients with complete or partial responses did not show subsequent reaccumulation of pleural effusion after pleurodesis. The chest tube remained in place for an average of 6.4 days, draining a mean of 2,854 mL. The main side-effects were fever and pain that were easily treated with nonsteroidal anti-inflammatory drugs. Pleurodesis using both OK-432 and doxorubicin showed high efficacy for controlling malignant pleural effusions caused by lung cancer. PMID- 15332396 TI - Sleep-related breathing disorders: impact on mortality of cerebrovascular disease. AB - The aim of the study was to analyse the impact of sleep-related breathing disorders in a 2-yr survival follow-up of patients with a first ever stroke or transient ischaemic attack. The study followed 161 patients. Complete neurological assessment was performed in order to determine cerebrovascular risk factors, functional disability, and parenchymatous and vascular localisation, as well as stroke subtype categorisation. A sleep study was carried out using a portable respiratory recording device, The entire cohort was followed over a mean period of 22.8 months. The main outcome event was death and time of survival since the neurological event. A multivariate Cox's model was estimated. The patients were ages 72+/-9 yrs (mean+/-SD), and had a body mass index of 26.6+/ 3.9 kg x m(-2) and apnoea/hypopnoea index (AHI) of 21.2+/-15.7. Overall, mortality occurred in 22 cases, and the survival rate was 86.3%. Vascular disease accounted for 63.6% of deaths. Multivariate analysis selected four independent variables associated with mortality: 1) age; 2) AHI, with an implied 5% increase in mortality risk for each additional unit of AHI; 3) involvement of the middle cerebral artery; and 4) the presence of coronary disease. In conclusion, the findings suggest that sleep-related breathing disorders are an independent prognostic factor related to mortality after a first episode of stroke. PMID- 15332397 TI - Automatic CPAP titration with different self-setting devices in patients with obstructive sleep apnoea. AB - Autotitrating continuous positive airway pressure (CPAP) devices automatically adjust the pressure according to upper airway obstructions. The aim of this study was to compare the treatment effects of different automatic CPAP devices (AutoSet, Horizon and Virtuoso) with conventional CPAP in patients with obstructive sleep apnoea independently of financial manufacturer support. Twelve male patients with obstructive sleep apnoea were submitted to a crossover study protocol with overnight polysomnography for 6 consecutive nights. After diagnostic polysomnography, the CPAP pressure was manually titrated. Over the next 4 nights, the patients were treated with any one of the three automatic CPAP devices or fixed CPAP in random order. The apnoea/hypopnoea index on the diagnostic night was 67.3+/-21.7 events h(-1), and was significantly reduced to 0.7+/-1.2, 3.0+/-2.9, 2.3+/-2.5 and 12.0+/-13.6 events x h(-1) with the fixed CPAP, AutoSet, Horizon and Virtuoso devices respectively. An apnoea/ hypopnoea index of <5 events h(-1), an indicator of optimal treatment, was achieved in all patients with fixed CPAP and in 10 patients using the Autoset and Horizon devices, but in only six of the 12 using the Virtuoso. The mean pressure was significantly lower with the AutoSet and Virtuoso devices, but not with the Horizon as compared to fixed CPAP. The maximum pressure was significantly higher with the Horizon. It is concluded that automatic continuous positive airway pressure devices produce a significant reduction in apnoea/hypopnoea index; however, there is considerable difference in the efficacy of the various devices. PMID- 15332398 TI - Performance vigilance task and sleepiness in patients with sleep-disordered breathing. AB - Altered vigilance performance has been documented in patients with sleep-related breathing disorders (SRBDs). Sleep fragmentation, sleepiness, respiratory disturbances and nocturnal hypoxaemia have been suggested as the pathogenesis of these deficits, yet it remains difficult to find a good correlation between performance deficits and the above factors. In the present study, which performance measure better characterised SRBD patients and the main factors implicated in these disturbances were examined. The study group consisted of 152 patients and 45 controls, all examined using a performance vigilance task and subjective sleepiness assessment. Speed and accuracy in the psychomotor vigilance task (PVT) were measured in patients and controls. Objective daytime sleepiness was assessed in the patient group using the maintenance of wakefulness test. In comparison with controls, PVT accuracy rather than speed seems to be affected in SRBD patients, with lapses and false responses significantly greater in patients with more severe objective sleepiness and higher apnoea/hypopnoea index. Although slowing and increased variability in reaction time were associated with shorter sleep latency in the maintenance of wakefulness test, subjective sleepiness, sleep fragmentation, nocturnal hypoxaemia and apnoea/hypopnoea index influenced mainly PVT accuracy. It is concluded that vigilance impairment, sleep fragmentation and severity of disease may partially and differentially contribute to the diurnal performance consequences found in sleep-related breathing disorders. Since the psychomotor vigilance task worsening is more marked in accuracy that in speed, measurement of lapses and false responses would better characterise the degree of diurnal impairment in these patients. PMID- 15332399 TI - Airway iron and iron-regulatory cytokines in cystic fibrosis. AB - Iron availability is critical to Pseudomonas aeruginosa. The current authors determined sputum iron, ferritin, microalbumin levels and total cell counts (TCC) in 19 adult patients with cystic fibrosis (CF) during an acute exacerbation and repeated analyses following a median of 12 days antibiotic treatment. The current authors also determined sputum interleukin (IL)-1beta and tumour necrosis factor (TNF)-alpha levels because of their putative role in intracellular iron homeostasis. Additional data were obtained from 17 stable CF patients, eight patients with stable chronic obstructive pulmonary disease (COPD) and six normal subjects. Overall, sputum iron, ferritin, microalbumin, IL-1beta and TNF-alpha concentrations and TCCs were significantly elevated in the CF patients compared to those with COPD and normal controls. Sputum ferritin levels were significantly elevated in acute versus stable CF patients and there was a trend for sputum TCC to be higher, but all other inflammatory indices were similar. In the CF patients, sputum iron was positively and strongly related to IL-1beta, TNF-alpha, ferritin and microalbumin levels, but negatively related to forced expiratory volume in one second % predicted. In those acute patients who clinically improved with antibiotics (n=14), there were significant decreases in sputum TCC, iron, ferritin and IL-1beta content, but not TNF-alpha or albumin levels. However, changes in sputum TNF-alpha in acute patients were still closely related to changes in iron, ferritin and albumin content, and changes in IL-1beta were related to changes in sputum ferritin content. Iron and iron-regulatory cytokines may play a role in cystic fibrosis lung disease and the increased iron content may even facilitate Pseudomonas aeruginosa infection. PMID- 15332400 TI - Umbilical cord trace elements and minerals and risk of early childhood wheezing and eczema. AB - It has been suggested that foetal nutrition might influence the inception of wheezing and atopic disorders in childhood but specific nutrients have not been implicated. In the Avon Longitudinal Study of Parents and Children umbilical cord samples were assayed for trace elements and minerals, and mothers were asked about wheezing and eczema in their children. Associations of cord concentrations of selenium, zinc, copper, manganese, magnesium, iron, lead and mercury with wheezing at 30-42 months, with wheezing patterns defined by the presence or absence of transient infant, later onset or persistent wheezing at 0-6 months and 30-42 months, respectively (n=2,044), and with eczema at 18-30 months (n=2,173), were analysed. Cord selenium was negatively associated with persistent wheeze (adjusted odds ratio (OR) per doubling concentration: 0.67). Cord iron was negatively associated with later onset wheeze (OR: 0.86) and with eczema (OR: 0.90). Children with high cord concentrations of selenium and iron were less likely than those with low concentrations to wheeze transiently in infancy. The level of foetal exposure to selenium and iron may possibly influence the risk of wheezing and eczema in early childhood although, in view of the multiple analyses carried out, it is possible that the main findings occurred by chance. PMID- 15332401 TI - Bronchiolitis obliterans syndrome in popcorn production plant workers. AB - Following sentinel case recognition, an excess of fixed airways obstruction was found among current workers in a microwave popcorn plant associated with butter flavouring exposures. In order to characterise the clinical presentation of sentinel cases, the medical records of sentinel cases were reviewed, interviews conducted and serial spirometric testing performed. Cases worked in microwave popcorn production, and five of the nine cases had mixed flavourings. Most had never smoked or smoked minimally. Cases showed onset of cough, shortness of breath and wheezing 5 months to 9 yrs after starting work at the popcorn plant. Initial forced expiratory volume in one second ranged 14.0-66.8% of the predicted value. Eight high-resolution computed tomography scans showed marked bronchial wall thickening and mosaic attenuation with air trapping. Open lung biopsy results were consistent with, or diagnostic of, constrictive bronchiolitis in two of three cases. Five cases are on lung transplantation waiting lists. After leaving employment, nearly all cases experienced stabilisation of their lung function within 2 yrs. Astute clinicians can help identify new causes of airways obstruction by alerting public health authorities to unexplained disease cases occurring in groups of workers. PMID- 15332402 TI - Acute mountain sickness is related to nocturnal hypoxemia but not to hypoventilation. AB - The purpose of the study was to investigate determinants of acute mountain sickness after rapid ascent to high altitude. A total of 21 climbers were studied ascending from <1,200 m to Capanna Regina Margherita, a hut in the Alps at 4,559 m, within <24 h. During their overnight stay at 4,559 m, breathing patterns and ventilation were recorded by calibrated respiratory inductive plethysmography along with pulse oximetry. In the following morning, acute mountain sickness was assessed. Altogether, 11 mountaineers developed pronounced symptoms of acute mountain sickness (Lake Louise score > or =5) and 10 did not (controls). Compared to controls, subjects with acute mountain sickness had lower nocturnal oxygen saturation (mean+/-SD 59+/-13% versus 73+/-6%), higher minute ventilation (7.94+/ 2.35 versus 6.06+/-1.34 L x min(-1)), and greater mean inspiratory flow, a measure of respiratory centre drive (0.29+/-0.09 versus 0.22+/-0.05 L x s(-1)). Periodic respiration was prevalent but not significantly different among the two groups (apnoea/hypopnea index 60.1+/-34.6 versus 47.1+/-42.6 events per h). The data suggest that pronounced nocturnal hypoxemia, which was not related to hypoventilation, may have promoted acute mountain sickness. Periodic breathing seems not to play a predominant role in the pathogenesis of acute mountain sickness. PMID- 15332403 TI - Effect of freezing of sputum samples on flow cytometric analysis of lymphocyte subsets. AB - Sputum samples should be processed shortly after induction to prevent cell degradation. For intermediate storage, freezing of homogenised samples or immediate fixation have been shown to be suitable for cytospins. The aim of this study was to investigate whether freezing or immediate fixation of sputum affect the analysis of lymphocyte subsets by flow cytometry. Selected plugs from 24 sputum samples were homogenised. One aliquot was processed immediately and analysed by flow cytometry. A second aliquot was homogenised, frozen at -20 C after addition of dimethylsulfoxide and stored for a median time of 6 days. In six samples a third aliquot was fixed in formalin after induction and stored for up to 72 h before further processing. Compared to immediate processing, percentages of total lymphocytes and T-suppressor cells were elevated after being frozen, with a minor decrease in the T4/T8 ratio. Proportions of total lymphocytes, T-helper and T-suppressor cells correlated between native and frozen samples, intra-class correlation coefficients being 0.74, 0.85 and 0.70, respectively. The formalin-fixed aliquots could not be analysed with the antibodies used. In conclusion, freezing seems to be a suitable technique to store sputum samples for flow cytometry of CD3, CD4 and CD8 lymphocyte subsets. Its effects were minor compared to the variation between subjects. PMID- 15332404 TI - New strategies to improve exercise tolerance in chronic obstructive pulmonary disease. AB - Breathlessness is the most common symptom limiting exercise in patients with chronic obstructive pulmonary disease (COPD). Exercise training can improve both exercise tolerance and health status in these patients, intensity being of key importance. Nevertheless, in these patients extreme breathlessness and/or peripheral muscle fatigue may prevent patients from higher levels of intensity. In this review article the literature concerning the different ways to optimise exercise tolerance in patients with COPD, with the objective of enhancing the tolerance to higher exercise training intensity, is summarised. Continuous positive airway pressure and different modalities of noninvasive positive pressure ventilation (NPPV) may reduce breathlessness and increase exercise tolerance in these patients. Respiratory muscle unloading and reduction in intrinsic positive end-expiratory pressure have been considered among mechanisms underlying these effects. Nevertheless, the role of NPPV in pulmonary rehabilitation, if any, is still controversial. The addition of nocturnal domiciliary NPPV during a daily exercise programme in patients with severe COPD resulted in an improvement in exercise tolerance and quality of life. In patients with severe COPD application of electrical stimulation combined with active limb mobilisation significantly improved muscle strength, and interval training has been shown capable of inducing physiological training effects. Oxygen supplementation in patients who do not desaturate during exercise seems to be the most promising treatment, since it allows for higher exercise intensities and, therefore, superior training efficacy. In conclusion, further studies are needed to define the appropriate patients in order to generalise such interventions. The modalities discussed should be used as adjuncts to a well designed comprehensive respiratory rehabilitation programme. PMID- 15332405 TI - Multiple system atrophy presenting as central sleep apnoea. AB - A 61-yr-old male presented with apparent idiopathic central sleep apnoea but after 4 yrs developed features of autonomic, cerebellar and extrapyramidal dysfunction consistent with a diagnosis of multiple system atrophy (MSA). Though central sleep apnoea can occur in multiple sleep apnoea, it is less frequent than obstructive sleep apnoea and occurs in the later stages of the disease. The pathogenesis of MSA involves gliosis and neuronal cell loss in specific areas of the central nervous system. Central sleep apnoea in MSA may be due to the depletion of cholinergic neurons in the arcuate nucleus of the medulla by apoptosis. This is the first description of multiple system atrophy presenting as central sleep apnoea. The current authors believe that multiple system atrophy should be considered in the differential diagnosis of late onset central sleep apnoea and progressive hypoventilation. PMID- 15332406 TI - A 7 month old presenting with respiratory distress. PMID- 15332407 TI - The role of omalizumab or rhuMAB-E25 in the treatment of allergic rhinitis. PMID- 15332408 TI - Alveolization: does "A" stand for appropriate morphometry? PMID- 15332409 TI - Pre- and post-bronchodilator spirometric values and the degree of reversibility in patients with COPD. PMID- 15332410 TI - Additive blockade of beta2-integrin adhesion of eosinophils by salmeterol and fluticasone propionate. PMID- 15332411 TI - Addressing musculoskeletal pain. PMID- 15332412 TI - Traumatic subungual hemorrhage in great toe. PMID- 15332413 TI - Orthostatic hypotension. A primary care primer for assessment and treatment. AB - Orthostatic hypotension (OH), defined as a decline in blood pressure when a person moves from a supine to sitting or standing position, is a common physical finding in the primary care setting. It is associated with several medical conditions and its prevalence increases with age. Treatment is specific to cause. Drug-induced OH often can be alleviated by reducing dosage or completely changing medications. OH secondary to autonomic insufficiency or neurogenic causes remains a challenge to manage, and a combination of non-pharmacologic and pharmacologic measures are needed. Recommendations are made for preventive measures, patient and caregiver education, and nonpharmacologic and pharmacologic approaches to treatment. Approaches to managing OH in conjunction with hypertension are also discussed. PMID- 15332414 TI - Osteoarthritis. A review of musculoskeletal aging and treatment issues in geriatric patients. AB - Musculoskeletal disease is extraordinarily prevalent among geriatric patients, with an incidence that is increasing with the growth of this segment of the population. It is a major cause of disability, dependency, dysmobility, pain, and depression. Rheumatologic evaluation of the geriatric patient provides challenges to the clinician in the proper interpretation of pain, the effect of comorbidities and multiple medications, and the need to consider an interdisciplinary approach that includes the recognition of cognitive deficits, functional evaluations, and treatment of muscle atrophy. Osteoarthritis accounts for 60% to 70% of joint disease in older persons, with pathologic and radiographic findings that are a "background" against which to consider pain and limitation of motion. Therapeutic strategies that incorporate reconditioning, the reduction of load-bearing, and adequate pain control may positively impact quality of life and reduce the potential for falls. PMID- 15332415 TI - Neuroleptic malignant syndrome. PMID- 15332416 TI - E-FALCONS. A mnemonic for monitoring the prescribing of atypical antipsychotics. PMID- 15332417 TI - Palliative nurse education: towards a common language. PMID- 15332418 TI - Is there such a thing as a good death? AB - The idea of a 'good death' is one which has been central to the palliative care movement but which in fact predates it. A number of recent articles have grappled with the concept. It is a subject which is difficult to quantify scientifically and this article explores the territory using a number of philosophical, theological, historical and literary sources. The changing meaning of the concept is traced through the premodern, modern and postmodern periods. In particular the influence on Western ideals about death of the two paradigmatic stories of the deaths of Socrates and Jesus are examined. It is argued that the dualistic thinking of Plato, which often underlies our thinking about death, is no longer adequate and the author offers the thinking of the Spanish philosopher Miguel de Unamuno as an alternative way of approaching our attitudes to death. Although the article does not seek to give a definitive answer to the question it raises, it suggests that thinking about good death should be broadened to accept the struggle with which many people face their death. PMID- 15332419 TI - Research methodology: cancer cachexia syndrome. AB - Cachexia is a syndrome and therefore does not have a specific definition. Patients are characterized by the presence of anorexia, early satiety, weight loss, weakness, anaemia and oedema. These features occur to a variable extent in different patients and may change in severity during the course of a patient's illness. The multifactorial origin of cachexia precludes a uniform pathophysiological definition. Taken together these factors have hindered clinical studies both at a fundamental level and in terms of the introduction of effective therapy. The advent of novel therapeutic targets (e.g., ubiquitin proteasome pathway) and biological response modifiers has opened possibilities for new clinical trials in cachexia. Regulatory authorities feel it is important not only to demonstrate efficacy in terms of patients' nutritional status (e.g., lean body mass) but also functional status (e.g., performance status). This article reviews current methods to assess the latter. Methods focused on measuring physical activity level (e.g., doubly labelled water technique or physical activity meters) promise objective data which can be readily interpreted in terms of clinically meaningful benefit. PMID- 15332420 TI - Systematic review of bisphosphonates for hypercalcaemia of malignancy. AB - BACKGROUND: Bisphosphonates are the treatment of choice for hypercalcaemia of malignancy (HCM) but there is no consensus regarding which drug or dose should be given. We designed a systematic review to investigate the efficacy of bisphosphonates in the treatment of HCM. METHODS: We identified randomized controlled trials (RCTs) by searching electronic databases, scanning of reference lists, and consultation with experts and pharmaceutical companies. Foreign papers were translated. Inclusion criteria were RCTs, confirmed malignant disease and measurement of serum calcium (ionized or corrected for albumin) postrehydration. The primary outcome was number of patients achieving normocalcaemia. Secondary outcomes were time to normocalcaemia, time to relapse and toxicity. RESULTS: Twenty-seven papers and two abstracts, using intravenous bisphosphonates, fulfilled the inclusion criteria. Data from 26 studies were used in analyses. Due to the heterogeneity of studies, meta-analysis could not be performed. Pamidronate was more effective than placebo, mithramycin, etidronate (7.5 mg/kg) and low-dose clodronate (600 mg), but equal to higher dose clodronate (1500 mg). Clodronate and etidronate were superior to placebo; incadronate was superior to elcatonin; gallium nitrate was superior to etidronate. No difference was seen between alendronate and clodronate. Three dose finding studies showed no difference between 30-90 mg of pamidronate, but one well designed study showed increasing efficacy with increasing dose. Studies using increasing doses of ibandronate (0.6-4 mg), alendronate (2.5-15 mg), and incadronate (2.5-10mg), showed a dose response. Duration of administration of pamidronate did not affect efficacy (six studies). CONCLUSION: Bisphosphonates normalize calcium in >70% patients with minimal side effects. Aminobisphosphonates are most effective at maintaining normocalcaemia and should be given in high dose irrespective of baseline serum calcium. PMID- 15332421 TI - Awareness of husband's impending death from cancer and long-term anxiety in widowhood: a nationwide follow-up. AB - BACKGROUND: We investigated the predictors and long-term consequences of awareness time - the length of time a woman is aware of her husband's impending death from cancer. METHODS: All women (n = 506) living in Sweden under 80 years of age who lost their husband/partner owing to cancer of the prostate in 1996 or of the urinary bladder in 1995 or 1996 were followed with an anonymous postal questionnaire, 2-4 years after their loss. RESULTS: We received completed questionnaires from 379 of the widows. Of these, 55 (15%) reported an awareness time of 24 hours or less, 56 (15%) of 3-6 months and 95 (26%) of one year or more. The associations between the awareness time and morbidity were of a reverted 'J-shape,' with awareness time of 24 hours or less carrying the highest risk and 3-6/6-12 months the lowest. On comparing the awareness time of 24 hours or less with 3-6 months (preformed response category), the relative risks for anxiety were found to be 1.9. (1.0-3.6) (visual digital scale) and 4.5 (1.0-20.0) for intake of tranquillising drugs. Those not informed of their husband's fatal condition or not provided with psychological support by caregivers during their husband's last months of life had an increased risk of a short awareness time. CONCLUSIONS: During a man's terminal cancer illness, the wife's awareness time varies considerably and is influenced by information and psychological support from caregivers. A short awareness time may result in an additional and avoidable psychological trauma. PMID- 15332422 TI - Goal setting as a measure of outcome in palliative care. AB - Setting goals and assessing outcomes are essential elements in palliative care. This paper describes a multiprofessional project, conducted under the auspices of clinical audit, which attempted to evaluate important outcomes of care. Over a six-month period there were 123 consecutive admissions to the hospice. These patients and their carers, as well as the staff, were encouraged to set explicit goals for, and evaluate outcomes of, their care. As anticipated, we encountered many difficulties in this, but there were benefits. We were able to record goals of admission from the patient in 97 cases (79%), their main carer in 74 cases (63%) and from hospice staff in 120 cases (98%). Patient and carer goals were often more functional and specific whereas the staff goals tended to be more problem or symptom focused. The achievement of these goals was evaluated by patients, carers and staff at discharge or death (where possible) with the majority being fully or partially met. Only 15 patients and 9 carers thought that some or all of their goals had not been achieved with just 4 recording that their goals had changed. Overall, it was a worthwhile (although time-consuming) exercise and, as a result of the 'audit', clear goals and outcomes from patient, carer and staff perspectives are now routinely recorded for all admissions to the hospice and are used to focus multiprofessional patient review. Having analysed the process as well as the results of the 'audit', we would encourage others not to be daunted from undertaking similar projects. PMID- 15332423 TI - Recruiting patients into a primary care based study of palliative care: why is it so difficult? AB - In the UK, researchers' access to study populations and control over selection of participants is becoming increasingly constrained by data protection and research governance legislation. Intervening stages placed between researchers and the population they wish to study can have serious effects on recruitment and ultimately on the validity of studies. In this paper we describe our experiences of gaining access to patients for a study of palliative care in primary care. Despite considerable time and resources dedicated to recruitment, a smaller than anticipated study sample was achieved. We found that gatekeeping by ethics committees and practitioner control over sample selection were significant hurdles in accessing patients for the study. Gatekeeping responsibilities represent considerable challenges for researchers seeking to obtain a representative study sample, not just in palliative care, but for research in general in health care. PMID- 15332424 TI - Older people's views about home as a place of care at the end of life. AB - OBJECTIVES: To explore the attitudes of older people towards home as a place of care when dying. DESIGN: A two-phase qualitative study using focus groups and semi-structured interviews. PARTICIPANTS: Eight focus group discussions were held with 32 participants recruited from six purposively selected community groups representing older people in Sheffield, UK. A further 16 men and 29 women participated in semi-structured interviews. RESULTS: Participants identified that home was more than a physical location, representing familiarity, comfort and the presence of loved ones. While participants anticipated that home would be their ideal place of care during dying, practical and moral problems associated with it were recognised by many. Some had no informal carer. Others did not want to be a 'burden' to family and friends, or were worried about these witnessing their suffering. Those who had children did not wish them to deliver care that was unduly intimate. Concerns were expressed about the quality of care that could be delivered at home, particularly in relation to accommodating health technologies and providing adequate symptom relief. Worries were also expressed about those living in poor material circumstances. Mixed views were expressed about the presence of professional carers within the home. Although they were seen to provide much needed support for the informal carer, the presence of 'strangers' was regarded by some as intrusive and compromising the ideal of 'home'. DISCUSSION: Older people perceive factors they associate with 'home' as crucial to a good death, most notably presence of friends and family, but many anticipate that they would prefer to be cared for elsewhere when dying. These findings run counter to assumptions that the medicalised, institutional death cannot be a 'good death'. It is important that dying in hospital is not demonized, but rather efforts made to examine how institutional deaths can take on a more meaningful quality. PMID- 15332425 TI - The end of life: informal care for dying older people and its relationship to place of death. AB - OBJECTIVE: This study examined the features of informal end-of-life care of older people living in the community and the association between informal care characteristics and dying at home. METHODS: Retrospective data were obtained from interviews and self-administered questionnaires of 56 persons who had been primary caregivers of older relatives in the last three months of their lives. RESULTS: Results showed that informal caregivers of terminally ill older people living in the community provided a considerable amount of personal, household, and management care. Secondary informal caregivers and formal caregivers assisted resident primary caregivers less often than nonresident primary caregivers. Primary caregivers who felt less burdened, who gave personal care more intensively, and/or who were assisted by secondary caregivers, were more likely to provide informal end-of-life care at home until the time of death. CONCLUSIONS: Our study showed that informal care at the end of life of older people living in the community is complex, since the care required is considerable and highly varied, and involves assistance from secondary informal caregivers, formal home caregivers as well as institutional care. Burden of informal care is one of the most important factors associated with home death. More attention is needed to help ease the burden on informal caregivers, specifically with regard to resident caregivers and spouses. Since these resident caregivers were disadvantaged in several respects (i.e., health, income, assistance from other carers) compared to nonresident caregivers, interventions by formal caregivers should also be directed towards these persons, enabling them to bear the burden of end-of-life care. PMID- 15332426 TI - Falls in hospices: a cancer network observational study of fall rates and risk factors. AB - Falls have been identified as a problem in frail patient populations, but their risk factors have not been studied prospectively in the hospice setting. We calculated fall rates in three hospices in the Yorkshire region, identified risk factors for falls based on previous studies in elderly patients, and then carried out a prospective study of inpatients in two of these hospices. We compared these risk factors in patients who subsequently fell with patients that did not fall during the study period. Information was recorded on 102 admissions. Twelve patients fell, generating 23 falls; six patients fell more than once. Significant risk factors for falling were cognitive impairment, low systolic lying and standing blood pressure, visual impairment and age over 80. Males with these risk factors fell more often than female patients with these risk factors. Strategies to prevent falls in hospice inpatients need to be directed appropriately towards patients with cognitive and visual impairment and low systolic blood pressure. PMID- 15332427 TI - Dying and defibrillation: a shocking experience. AB - Palliative Care physicians are frequently involved in the care of patients with significant comorbidity and often have to take coexisting conditions into account when treating patients. An example of an area in which this is particularly relevant and will undoubtedly increase is presented with the case report of a patient with terminal metastatic lung carcinoma and an Implantable Cardioverter Defibrillator (ICD) in place. The role of the ICD in preventing the patient from dying comfortably is discussed, as are means of deactivating the device. We conclude that patients with ICDs and terminal disease should have the issue of deactivation addressed at the earliest possible opportunity as practical difficulties may arise in the emergency setting, especially in the nonhospital environment. PMID- 15332428 TI - Approval by MREC of a modified patient information and consent form. does this set a precedent for trials in palliative care? PMID- 15332429 TI - Mother-to-child transmission of HIV-1: advances and controversies of the twentieth centuries. AB - Mother-to-child transmission (MTCT) is the overwhelming source of HIV-1 infection in young children. According to the World Health Organization (WHO), during the year 2003, despite effective antiretroviral (ARV) therapy, there were approximately 700,000 new infections in children worldwide, the majority of whom were from resource-limited countries. Alternative protocols to the long-course and complex regimens of ARV drugs, which in high-income countries have almost eradicated HIV MTCT, have been shown to reduce early transmission rates by 38 50%. However, the accumulation of drug resistance and the long-term toxicities of ARVs mean that alternative approaches need to be developed. Furthermore, transmission via breastfeeding, which accounts for one third of all transmission events, can reduce the benefits of short-course therapies given to women for the prevention of MTCT. The complex mechanisms and determinants of HIV-1 MTCT and its prevention in the different routes of transmission are still not completely understood. Despite the large contribution that many international agencies have made during the past 10-15 years in support of observational and intervention trials, as well as basic scientific research, HIV-1 MTCT intervention trials and basic research often are not integrated, leading to the generation of a fragmented picture. Maternal RNA levels, CD4+ T-cell counts, mode of delivery and gestational age were shown to be independent factors associated with transmission. However, these markers are only partial surrogates and cannot be used as absolute predictors of MTCT of HIV-1. Studies on the role of viral characteristics, immune response and host genomic polymorphisms did not always achieve conclusive results. Although CCR5-using viruses are preferentially carried by HIV-1 infected women as well as transmitted to their infants, the 32 basepair deletion of the CCR5 gene was not shown to influence perinatal MTCT. X4 viruses are apparently hampered in MTCT, although transmission of syncytium inducing (SI) viruses, which use CXCR4, can occur when the mother carries such virus. Recently, there has been evidence of multiple virus variant transmission during peripartum MTCT. If viral escape from cytotoxic T-lymphocyte (CTL) recognition was repeatedly detected in transmitting mothers, no conclusive results were obtained on the role of the humoral immune response. The hypothesis on the mechanisms of selection during MTCT are still an open question, and include possibly that the transmitted variant is derived from a variant in the mother that escaped immune response, or that transmission is a stochastic event with the random transmission of a limited number of viral variants, or otherwise that selection occurs in the infant through a replication advantage of some transmitted viral variants. Although global access to ARV therapy certainly remains the primary goal to achieve the immediate reduction of MTCT of HIV-1, it is also evident that new and additional innovative strategies are needed. PMID- 15332430 TI - The role of CD8+ T-cell response in HIV infection. AB - CD8+ T-cells with cytotoxic (CTL) activity play a pivotal role in controlling viral infections. Although most patients chronically infected with HIV have CTL response against the virus, for reasons that are not well understood this response is not able to successfully control viral replication. The crucial role of this type of response has been clearly demonstrated in the setting of acute infection using the simian model of AIDS, in which a strong CTL response develops, supporting its role in humans. This approach has been possible due to the development of new assays to quantify CTL activity with great sensitivity and specificity. The interaction of CTL response and HIV during this acute stage of infection is crucial, since it most probably determines the viral set-point and thus the rate of HIV disease progression. In the setting of chronic HIV infection, the use of tetrameric complexes and IFN-gamma production assays have made it possible to investigate the different functional aspects of these cells and have also facilitated the evaluation of this response in large patient populations. Defects in cytokine production and in perforin expression have been found, as well as alterations in phenotypic maturation and a low proliferation of these cells. All these findings have been cited to explain the inability of CTLs to efficiently control virus replication. Nonetheless, accumulating evidence points toward an important role for CTL response in the partial containment of HIV replication in chronic infection. An especially strong support for this observation derives from studies analyzing the selective pressure exerted by the immune response over viral evolution. Very recently, longitudinal and cross sectional studies in large populations of patients have demonstrated that viral evolution is in part driven by HIV-specific T-cell responses. PMID- 15332431 TI - Progression of HIV to AIDS: a protective role for HLA-B27? AB - HLA-B27 is known for its strong association with inflammatory spondyloarthropathies (SpA), a group of rheumatic diseases. Apart from playing its role in the onset of these inflammatory diseases, HLA-827 is so ubiquitous in the world that the carrying of this gene must have also have an advantage. There are some indications that a beneficial effect can be found as a less severe course of viral infections among B27-carriers. The literature on this subject was reviewed and revealed a favorable course of infection with influenza virus, herpes simplex type 2 virus, Epstein-Barr virus and, even more interesting, a protective effect of HLA-B27 in the progression of HIV infections. The course of HIV infection differs among individuals and is thought to be partly related to host-factor variability, reflecting broad genetic heterogeneity. The polymorphic human leukocyte antigens (HLA) are herein analyzed intensively with respect to this relationship. Cytotoxic T lymphocyte (CTL) responses, activated by HLA antigen presentation, are implicated in the control of HIV replication. An immunological explanation for the protective role for HLA B27 in HIV disease is that B27+ patients have a specific and strong CTL response against the p24 epitope, a conservative HIV protein that does not easily mutate. Some HLA genes seen in long-term non-progressors (LTNP) (>10 years disease free) are associated with a favorable prognosis. One of the alleles found predominantly in LTNPs is HLA-B27. More genetic factors seem to influence disease progression in HIV infections. Therefore, it would be interesting to further explore the influence of the genetic make up of these HIV-infected individuals. Knowledge of the immunogenetic profile might give clues for the individual course of the HIV infection, may influence the development of drug-resistant viruses and will possibly lead to a tailored therapeutic strategy in HIV-infected persons. PMID- 15332432 TI - Endocarditis in the setting of HIV infection. AB - BACKGROUND: Cardiac complications are becoming more important in patients with HIV infection. The most common is infective endocarditis (IE) in patients who are intravenous drug addicts (IVDA). Other less common problems are pulmonary hypertension, cardiotoxicity, pericardial effusion, cardiac neoplasms, etc. PATIENTS AND METHODS: A literature review of published studies on IE was done and the personal experience of the authors is reflected. RESULTS: The clinical pattern of IE has remained unchanged. It is usually due to Staphylococcus aureus and is more commonly localized to the right side of the heart. It is not clearly defined if HIV infection is responsible for the worst evolution in these patients and the treatment is the same as that used in HIV- subjects. CONCLUSIONS: IE is responsible for 5-20% of hospital admissions and for 5-10% of total deaths in IVDA patients with HIV infection, but the clinical outcome of the patients depends on the affected valve and the culture germen rather than the HIV serostatus. PMID- 15332433 TI - 2004: which HIV-1 drug resistance mutations are common in clinical practice? AB - The emergence of drug resistance remains a major problem for the treatment of HIV infected patients. However, the variety of mutational patterns that evolve in clinical practice have made the application of resistance data to clinical decision-making challenging. Despite (or because of) an abundance of drug resistance data from disparate sources, there is only limited information available describing the patterns of drug resistance which usually appear in the clinic. Here we attempt to address this issue by reviewing HIV drug resistance in the population of patients failing antiretroviral therapy in British Columbia, Canada from June 1996 to December 2003 as an example. Our findings suggest that, although hundreds of mutations have been associated with resistance, relatively few key mutations occur at a high frequency. For example, only the nucleoside reverse transcriptase inhibitor (NRTI) mutations M184V, M41L T215Y, D67N, K70R and L210W, non-nucleoside reverse transcriptase inhibitor (NNRTI) mutations K103N and Y181C, and protease inhibitor (PI) mutation L90M, occur in more than 10% of samples tested for resistance in this population. The introduction of new drugs allows for the selection of new mutations. Trends in the prevalence of resistance associated mutations have generally followed trends in drug usage, but have not always mirrored them. The phenomenon of cross-resistance can play an important role in the efficacy of new antiretroviral agents, even before they become available. The extent of this cross-resistance depends in part on the prevalence of specific mutations in the population of individuals who have previously received antiretroviral therapy. Hence there is a need to determine which mutations are prevalent in the treated population. The tremendous capacity of HIV to adapt means that common resistance pathways are likely to change over time, and new pathways to resistance are likely to continue to be discovered in the future. PMID- 15332434 TI - Developing mental imagery using a digital camera: a study of adult vocational training. AB - This study was undertaken to explore the use of a digital camera for mental imagery training of a vocational task with two young adult men with Down syndrome. The results indicate that these particular men benefited from the use of a collaborative training process that involved mental imagery for learning a series of photocopying operations. An outline of a mental imagery assessment method is provided along with a description of the training procedures that were used in the study. Mental imagery was taught and rehearsed by means of a Powerpoint computer presentation. Trainers and participants worked collaboratively to complete a task analysis and to take photos of each person performing the operations correctly. Through the use of self-modelling and feed forward, participants were able to learn from observing their own actions and, in particular, to learn from 'successes' that they have not yet had. On the basis of this pilot study, it is proposed that mental imagery training is an important new approach for collaborative training, especially for individuals whose language systems are not well developed. There is a need, however, for further investigation into the role of mental imagery as this relates to memory, self regulation and metacognition. PMID- 15332435 TI - Mental rotation abilities in individuals with Down syndrome--a pilot study. AB - This pilot study was designed to examine mental rotation ability in individuals with Down syndrome. 7 individuals with Down syndrome (mean mental age = 8.18 +/- 2.73 years; mean chronological age = 29.8 +/- 5.4 years) and a group of 9 typically developing children, matched for mental age, (mean mental age = 8.40 +/ 1.73 years; mean chronological age = 7.2 +/- 1.2 years) were given a version of Cooper and Shepherd's (1973) mental rotation paradigm. On each trial, participants viewed a symbol representing an upper case 'F' or a mirror image of an 'F'. The symbol was presented at one of eight different orientations. The participant's task was to determine whether the letter was reversed or non reversed. Interestingly, both groups showed similar trends in increased reaction times with increasing angular disparity, suggesting that both groups were performing mental rotations. There was no significant difference in reaction time between the typically developing and Down syndrome groups, however, the Down syndrome group made significantly more errors than the typically developing group. Participants with Down syndrome were able to carry out the mental rotations at well above chance level and mental rotation ability was shown to correlate with mental age. PMID- 15332436 TI - A comparison between older persons with down syndrome and a control group: clinical characteristics, functional status and sensorimotor function. AB - The increase in life expectancy within the general population has resulted in an increasing number of elderly adults with intellectual disability, and this is reflected in the increased life expectancy in persons with Down syndrome, currently about 56 years. The aim of this study was to study the clinical characteristics, the functional status and sensori-motor function of 10 older persons with Down syndrome (mean age 59 years), 13 younger persons with Down syndrome (mean age 44 years) and compare them with 38 adults with intellectual disability without Down syndrome and a control group of people without intellectual disability. All the persons with Down syndrome and intellectual disability resided in two residential living centres in Israel, while the 31 older persons without intellectual disability (mean age 75 years), who served as the control group, lived in an independent living facility. The study considered demographic data, medical backgrounds, physical and functional tests. The results showed that the older persons with Down syndrome in the study were more obese, shorter and had more medical problems than both the older persons with intellectual disability and the control group. The functional performance of the older adults with Down syndrome was more impaired in comparison with both other groups. It is postulated that their slower responses may be explained by a less physically active lifestyle, that may accelerate the onset of disease, resulting in symptoms associated with aging that are detrimental to health. PMID- 15332437 TI - What's it all about? Investigating reading comprehension strategies in young adults with down syndrome. AB - The purpose of reading is for the reader to construct meaning from the text. For many young adults with Down syndrome, knowing what the text is all about is difficult, and so for them the activity of reading becomes simply the practice of word calling. It is suggested in the literature that for those individuals with Down syndrome, learning can continue into adolescence and that this may be the optimal time for learning to occur. However, a review of the literature revealed limited empirical research specifically relating to the reading comprehension of young adults with Down syndrome. Recent findings from Latch-On (Literacy And Technology Hands On), a research-based literacy and technology program for young adults with Down syndrome at the University of Queensland, revealed that comprehension remained the significant area of difficulty and showed least improvement (Moni & Jobling, 2001). It was suggested by Moni and Jobling (2001) that explicit instruction in comprehension using a variety of strategies and meaningful, relevant texts was required to improve the ability of young adults with Down syndrome to construct meaning from written texts. This paper is based on an action research project that was developed within the Latch-On program. The project utilised a modification of Elliot's (1991) action research model and was conducted to investigate specific teaching and learning strategies that would enhance the reading comprehension of young adults with Down syndrome. The participants were 6 young adults with Down syndrome ranging in age from 18 to 25 years. As the data from this project are still being analysed, preliminary findings of one participant are presented as a case study. The preliminary findings appear to indicate that the program of specific teaching and learning reading comprehension strategies used in this project was beneficial in the participant's reading comprehension. PMID- 15332438 TI - Parental experiences of feeding problems in their infants with Down syndrome. AB - Although feeding difficulties in infants with Down syndrome are described in the literature, the experiences of parents regarding the feeding problems of their infants are largely omitted. In order to promote closer collaboration with families and speech-language therapists in early communication intervention, the study investigated some experiences of a group of parents concerning the feeding problems of their infants with Down syndrome. A descriptive survey approach was implemented to collect quantitative data. The results revealed the type of feeding problems and associated conditions occurring in the infants of the twenty participants; the type of feeding methods used; the assistance received during the feeding problems; the emotions experienced in relation to the feeding problems in their infants as well as suggestions made by the participants regarding the management of early feeding problems. Numerous feeding problems occurred in the infants and the variety of emotions experienced by the participants indicated that feeding therapy and counseling were required. As a result of the different conditions underlying the feeding problems of the participants' infants, a comprehensive approach, which included therapeutic and medical intervention, was necessary. In order to provide effective early communication intervention services from birth, the results indicated that the knowledge, skills and sensitivity of speech language therapists to identify caregiver needs, to provide appropriate family-focused intervention and to make recommendations regarding the management of feeding problems in infants with Down syndrome is of great importance. PMID- 15332439 TI - Hair zinc level in Down syndrome. AB - Immunological, endocrinological, and haematological abnormalities are relatively common in people with Down syndrome (Cuadrado & Barrena, 1996; Decoq & Vincker, 1995; Hestnes et al., 1991; Sustrova & Strbak, 1994; Nespoli, Burgio, Ugazio & Maccario, 1993; Kempski, Chessells & Reeves, 1997; Kivivuori, Rajantie, & Siimes, 1996; David et al., 1996; Gjertson, Sturm & Berger, 1999). Zinc is one of the elements that act in the maintenance of normal function of these systems. This study was designed to investigate zinc levels in children with Down syndrome. Zinc levels were measured in hair using atomic absorption spectrophotometry. The hair zinc level of 19 children with Down syndrome was compared with the zinc level of 11 typically developing children. Hair zinc levels were found to be significantly lower (p < .05) in those with Down syndrome (average 95.18 +/- 56.10 ppm) than in the typically developing children (average 208.88 +/- 152.37 ppm). Some of the problems experienced by children with Down syndrome may be due to these low zinc levels, but further research is required to confirm these results, and to establish any correlation with these problems. PMID- 15332440 TI - Unique self-sustaining circadian oscillators within the brain of Drosophila melanogaster. AB - In Drosophila circadian rhythms persist in constant darkness (DD). The small ventral Lateral Neurons (s-LNv) mainly control the behavioral circadian rhythm in consortium with the large ventral Lateral Neurons (l-LNv) and dorsal Lateral Neurons (LNd). It is believed that the molecular oscillations of clock genes are the source of this persistent behavior. Indeed the s-LNv, LNd, Dorsal Neurons (DN)-DN2 and DN3 displayed self-sustained molecular oscillations in DD both at RNA and protein levels, except the DN2 oscillates in anti-phase. In contrast, the l-LNv and DN1 displayed self-sustained oscillations at the RNA level, but protein oscillations quickly dampened. Having self-sustained and dampened molecular oscillators together in the DN groups suggested that they play different roles. However, all DN groups seemed to contribute together to the light-dark (LD) behavioral rhythm. The LD entrainment of LN oscillators is achieved through Rhodopsin (RH) and Cryptochrome (CRY). CRY's expression in all DN groups implicates also its role in LD entrainment of DN, like in DN1. However, mutations in cry and glass that did not inflict LD synchronization of the DN2, DN3 oscillator implicate the existence of a novel photoreceptor at least in DN3. PMID- 15332441 TI - A locus for circadian period of locomotor activity on mouse proximal chromosome 3. AB - Lengthened circadian period of locomotor activity is a characteristic of a congenic strain of mice carrying a nonsense mutation in exon 5 of the carbonic anhydrase II gene, car2. The null mutation in car2 is located on a DBA/2J inbred strain insert on proximal chromosome 3, on an otherwise C57BL/6J genomic background. Since reducing the size of the congenic region would narrow the possible candidate genes for period, two recombinant congenic strains (R1 and R2) were developed from the original congenic strain. These new congenic strains were assessed for period, genetic composition, and the presence of immunoreactive carbonic anhydrase II. R1 mice were homozygous DBA/2J for the distal portion of the original DBA/2J insert, while R2 mice were homozygous DBA/2J for the proximal portion. R1 mice had a significantly lengthened period compared to R2 mice and wild-type C57BL/6J mice, indicating that the gene(s) affecting period is likely found within the reduced DBA/2J insert (approximately 1 cM) in the R1 mice. The R1 mice also possessed the null mutation in car2. This study confirmed the presence of a gene(s) affecting period on proximal chromosome 3 and significantly reduced the size of the congenic region and the number of candidate genes. Future studies will focus on identifying the gene influencing period. PMID- 15332442 TI - Effects of temperature, photoperiod, and light intensity on the eclosion rhythm of the high-altitude Himalayan strain of Drosophila ananassae. AB - Eclosion rhythm of the high-altitude Himalayan strain of Drosophila ananassae from Badrinath (altitude 5123 m) was temperature-dependent and at 21 degrees C, it was entrained by cycles of 12h light: 12h darkness (LD 12:12) and free-ran in constant darkness, however, it was arrhythmic at 13 degrees C or 17 degrees C under identical experimental conditions (Khare, P. V., Barnabas, R. J., Kanojiya, M., Kulkarni, A. D., Joshi, D. S. (2002). Temperature dependent eclosion rhythmicity in the high altitude Himalayan strains of Drosophila ananassae. Chronobiol. Int. 19:1041-1052). The present studies were designed to see whether or not these strains could be entrained at 13 degrees C, 17 degrees C, and 21 degrees C by two types of LD cycles in which the photoperiod at 100 lux intensity varied from 6h to 18h, and the light intensity of LD 14:10 cycles varied from 0.001 lux to 1000 lux. All LD cycles entrained this strain at 21 degrees C but not at 13 degrees C or 17 degrees C. These results demonstrate that the entrainment of eclosion rhythm depends on the ambient temperature and not on the photoperiod or light intensity of LD cycles. Thus the temperature has taken precedence over the light in the entrainment process of eclosion rhythm of the high altitude Himalayan strain of D. ananassae. This may be the result of natural selection in response to the environmental temperature at Badrinath that resembles that of the sub-Arctic region but the photoperiod or light intensity are of the subtropical region. PMID- 15332443 TI - Wavelength dependency of light-induced effects on photoperiodic clock in the migratory blackheaded bunting (Emberiza melanocephala). AB - The effects of light wavelength on photoperiodic clock were determined in the migratory male blackheaded bunting (Emberiza melanocephala). We constructed an action spectrum for photoperiodic induction (body fattening, gain in body mass, and gonadal recrudescence) by exposing birds for 4.5 weeks to 13 h light per day (L:D = 13:11 h) of white (control), blue (450 nm), or red (640 nm) color at irradiances ranging from 0.028 to 1.4Wm(-2). The threshold light irradiance for photoinduction was about 10-fold higher for blue, compared to red and white light. Phase-dependent effects of light wavelength on the photoperiodic clock were further examined in the next two sets of skeleton photoperiods (SKPs). In the first set of SKPs, birds were exposed for four weeks to asymmetrical light periods (L:D:L:D= 6:6:1:11 h) at 0.25+/-0.01 W m(-2); two light periods applied were of the same (450nm: blue:blue, B:B; 640nm, red:red, R:R) or different (blue:red, B:R or red:blue, R:B) wavelengths, or of white:white (W:W, controls). Photoperiodic induction occurred under R:R and B:R, but not under B:B and R:B light conditions; the W:W condition induced an intermediate response. The second set of SKPs used symmetrical light periods (L:D:L:D = 1:11:1:11 h), and measured effects also on the activity rhythm. Birds were first exposed to one of the four SKPs (R:R, B:B, R:B, or B:R) for three weeks, subsequently were released into dim constant light (LLdim; approximately 0.01 Wm(-2), the night light used in an L:D cycle) for two weeks, and then were returned to respective SKPs for another three weeks. Activity was greater in the R:R compared to B:B, and in B:R compared to R:B light condition. Zugunruhe (intense nighttime activity, indicating migratory restlessness in a caged situation) developed under the R:R and B:R, but not the B:B and R:B, light condition. Under LLdim, all birds free-ran with a period >24h, the Zugunruhe had a circadian period longer than the daytime activity, and the re entrainment to SKPs was influenced by the position of light periods relative to circadian phase of the activity rhythm. Photoperiodic induction at the end of 8 weeks was found in the R:R and B:R, but not in B:B, light conditions; in the R:B condition only one bird had initiated testes. Taken together, these results suggest that in the blackheaded bunting, the circadian photoperiodic clock is differentially responsive to light wavelengths; this responsiveness is phase dependent, and the development of Zugunruhe reflects a true circadian function. Wavelength-dependent response of the photoperiodic clock could be part of an adaptive strategy in evolution of the seasonality in reproduction and migration among photoperiodic species under wild conditions. PMID- 15332444 TI - Reliability of the circadian rhythm of water and macronutrient-rich diets intake in dietary choice. AB - Rats with ad libitum water and the ability to self-select among three macronutrient-rich diets--carbohydrate (CHO), protein (PRO), and lipid (LIP)- show a circadian rhythmicity in their ingestion. The aim of the present study was to determine whether this circadian rhythmicity is reliable from day to day. Eight rats were offered ad libitum water and a choice of three isoenergetic diet rations providing carbohydrate, protein, and lipid. Water and food intake was recorded every 3 h for 7 days. The reliability of the circadian rhythm of water and food intake was assessed by the Intraclass Correlation Coefficient (ICC) and the test-retest reliability using the Pearson's Correlation Coefficient (r). The results showed that the circadian rhythm of water, CHO, and PRO intake are strongly reliable. However, the circadian rhythm of LIP intake is less reproducible. Among the three reliable parameters-water, CHO, and PRO, the circadian rhythm of water intake was the most reproducible over 7 days. This suggests that water intake may be used as a marker of circadian rhythmicity in ingestive behavior. PMID- 15332445 TI - 24-hour changes in circulating prolactin, follicle-stimulating hormone, luteinizing hormone, and testosterone in young male rats subjected to calorie restriction. AB - This work analyzes the effect of calorie restriction on the 24h variation of pituitary-testicular function in young male Wistar rats by measuring the circulating levels of prolactin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone. Control animals were provided an equilibrium calorie diet and the experimental animals a calorie-restriction diet equivalent to 66% of food restriction for four weeks starting on day 35 of life. Different groups of control and experimental rats were killed at 6h intervals around the clock, beginning I h after light on (HALO). Compared to the control animals, the mean secretion of prolactin was augmented and that of LH and testosterone decreased in calorie-restricted rats, whereas FSH release remained unchanged. Significant changes in the 24 h secretory pattern of circulating prolactin, LH, and testosterone occurred in the calorie-restricted rats. These include the appearance of a second maximum of plasma prolactin at 21 HALO, blunting of the LH peak seen at 13 HALO, and phase-shift of the testosterone peak from 13 HALO in controls to 17 HALO in calorie-restricted rats. The significant positive correlation between individual LH and testosterone levels found in controls was no longer observed in calorie-restricted rats. Availability of nutrients presumably affects the mechanisms that modulate the circadian variation of the pituitary-gonadal axis in growing male rats. PMID- 15332446 TI - Influence of fasting and exercise on the daily rhythm of serum leptin in the horse. AB - The hormone leptin is secreted by white adipocytes and regulates food intake and energy expenditure in rodents and humans. The goal of the present study was to investigate the existence of a daily rhythm of serum leptin in horses and its dependence on fasting and physical exercise. A robust daily rhythm of leptin was found in both athletic and sedentary horses, with a daytime trough and a peak in the dark phase. While physical exercise never induced changes in circulating leptin, fasting reliably affected serum leptin levels. Food deprivation did not abolish the daily rhythm of serum leptin, but daily mean leptin levels in fasted horses were significantly lower than in regularly fed horses. This result indicates that leptin production is not a mere consequence of feeding behavior. The fact that in a large animal such as the horse a short fast decreases leptin without significantly changing the body weight demonstrates that changes in levels of circulating leptin associated with food restriction do not solely reflect changes in amount of body fat. PMID- 15332447 TI - Prolonged mild hypoxia modifies human circadian core body temperature and may be associated with sleep disturbances. AB - Fatigue is often reported after long-haul airplane flights. Hypobaric hypoxia, observed in pressurized cabins, may play a role in this phenomenon by altering circadian rhythms. In a controlled cross-over study, we assessed the effects of two levels of hypoxia, corresponding to cabin altitudes of 8000 and 12,000 ft, on the rhythm of core body temperature (CBT), a marker of circadian rhythmicity, and on subjective sleep. Twenty healthy young male volunteers were exposed for 8 h (08:00-16:00 h) in a hypobaric chamber to a cabin altitude of 8000 ft and, 4 weeks later, 12,000 ft. Each subject served as his own control. For each exposure, CBT was recorded by telemetry for two 24h cycles (control and hypoxic exposure). After filtering out nonphysiological values, the individual CBT data were fitted with a five-order moving average before statistical group analysis. Sleep latency, sleep time, and sleep efficiency were studied by sleep logs completed every day in the morning. Our results show that the CBT rhythm expression was altered, mainly at 12,000 ft, with a significant increase of amplitude and a delay in the evening decline in CBT, associated with alterations of sleep latency. Mild hypoxia may therefore alter circadian structure and result in sleep disturbances. These results may explain in part the frequent complaints of prolonged post-flight fatigue after long flights, even when no time zones are crossed. PMID- 15332448 TI - Morningness-eveningness and lifestyle regularity. AB - The relation between morningness (M) - eveningness (E) and lifestyle regularity was studied in a convenience sample of 100 healthy subjects aged between 20 and 59 yrs (47 males and 53 females; mean age 33.6 yrs). Morningness-eveningness was measured by a single administration of the 13-item Composite Scale for Morningness (CSM). Lifestyle regularity was measured by requiring subjects to complete a five-item Social Rhythm Metric diary (SRM-5) each evening for two weeks. Each week of SRM-5 was analyzed separately and the two SRM scores averaged to yield the lifestyle regularity measure for the subject. Subjects were categorized by morningness into top and bottom quartiles of CSM score (denoted M types and E-types, respectively), with the remaining 50% of subjects denoted as intermediate (I-types). Mean SRM scores significantly differed between the three morningness groups (p <0.001) in the order E-types (SRM = 3.6), then I-types (SRM=4.0), then M-types (SRM=4.9), with higher scores indicating greater daily lifestyle regularity. The effect size of the E-type versus M-type difference was approximately 1.3. The relationship between the CSM and SRM scores was also confirmed using a correlational analysis (rho = 0.428; p < 0.001). This correlational finding was substantially weaker when age was partialed out (r=0.186; p=0.065), although there was still a trend toward a significan relationship. Thus it appears that morning types appear to be more regular in their daily lifestyle than are evening types, suggesting a relationship between these two aspects of human circadian behavior. PMID- 15332449 TI - Lack of evidence for a marked endogenous component determining food intake in humans during forced desynchrony. AB - In an attempt to investigate the relative importance of endogenous and exogenous factors in determining food intake, 14 healthy subjects were studied while living in an Isolation Unit (where external time cues were absent) for eighteen 28 h "days" (equal to 21 solar days). The subjects were free to spend their waking time as they chose, and they had a free choice of what they ate and when they ate it. The only restrictions were that no naps were allowed in the "daytime," that some time was required to perform a variety of tests at regular intervals throughout the 18.67 h waking periods, and that any food preparation had to be performed by the subjects themselves. Core (rectal) temperature and activity were monitored throughout, and the subjects answered a questionnaire on their eating habits at 3 h intervals during the waking periods. The questionnaire investigated reasons for eating or not eating a meal during the previous 3 h and, if a meal had been eaten, its type, the factors influencing that choice, and the subjects' subjective responses (hunger before, enjoyment during, and satiety after) to it. The results were analyzed (two-way ANOVA) in terms of both the imposed day length (the exogenous component) and the free-running period of the temperature rhythm (the endogenous component). Results indicated that by far the main reason for eating/not eating was hunger/lack of hunger rather than factors such as food availability and time-pressure. There were statistically significant effects of time within the imposed waking periods upon the type of meal eaten--"breakfast" tending to be a snack, "lunch" a small hot meal, and the "evening meal" a large hot meal. Hot meals (whether small or large) were associated with more hunger before the meal, more enjoyment of the meal, and a greater degree of satiety afterward than were cold meals. These effects suggest that the individuals adjusted their eating habits to fit in with the imposed wake times. By contrast, the effect of circadian phase upon food intake, the type of meal eaten, and subjective responses to the meal was much weaker, and either statistically nonsignificant (P > 0.10) or only marginally so (0.10 > P > 0.05). For example, a large hot meal was chosen as readily for an "evening meal," and subjective responses to it were the same, at whatever circadian phase it was eaten. We conclude that food intake during forced desynchronization is dominated by the waking schedule rather than by circadian influences; some of the implications of these findings when eating habits and the metabolism of food are concerned, particularly in night workers, are considered briefly. PMID- 15332450 TI - Comparison of parameters from rhythmometric models with multiple components on hybrid data. AB - Population multiple components is a statistical tool useful for the analysis of time-dependent hybrid data. With a small number of parameters, it is possible to model and to predict the periodic behavior of a population. In this article, we propose two methods to compare among populations rhythmometric parameters obtained by multiple component analysis. The first is a parametric method based in the usual statistical techniques for comparison of mean vectors in multivariate normal populations. The method, through MANOVA analysis, allows comparison of the MESOR and amplitude-acrophase pair of each component among two or more populations. The second is a nonparametric method, based in bootstrap techniques, to compare parameters from two populations. This test allows one to compare the MESOR, the amplitude, and the acrophase of each fitted component, as well as the global amplitude, orthophase, and bathyphase estimated when all fitted components are harmonics of a fundamental period. The idea is to calculate a confidence interval for the difference of the parameters of interest. If this interval does not contain zero, it can be concluded that the parameters from the two models are different with high probability. An estimation of p-value for the corresponding test can also be calculated. Both methods are illustrated with an example, based on clinical data. The nonparametric test can also be applied to paired data, a special situation of great interest in practice. By the use of similar bootstrap techniques, we illustrate how to construct confidence intervals for any rhythmometric parameter estimated from population multiple components models, including the orthophase, bathyphase, and global amplitude. These tests for comparison of parameters among populations are a needed tool when modeling the nonsinusoidal rhythmic behavior of hybrid data by population multiple component analysis. PMID- 15332451 TI - Time-of-day effects on anaerobic muscular power in a moderately warm environment. AB - This study evaluated the influence of a neutral vs. a moderately warm environment on the diurnal variation in muscular power. Twelve male subjects [27.0 (+/-4) years] performed two different jump tests [a squat jump (SJ) and a counter movement jump (CMJ)] and a brief maximal sprint on cycle ergometer (CS) in four different conditions (morning/neutral, morning/moderately warm and humid, afternoon/neutral, and afternoon/moderately warm and humid). The morning experiments were conducted between 07:00 and 09:00 h, and the afternoon experiments were conducted between 17:00 and 19:00 h. The mean laboratory temperatures and humidity were 20 (+/-1) degrees C, 70 (+/-5)% and 29 (+/-1) degrees C, 57 (+/-4)% for the neutral and moderately warm and humid conditions, respectively. Rectal temperature and leg skin temperature were significantly dependent on both time-of-day and ambient temperature. An interaction effect (P < 0.05) was noted between time-of-day and ambient temperature for the power developed for the CMJ, the SJ, and half of a pedal revolution during the cycling sprint. In summary, (i) the same subjects were influenced by time-of-day differently, depending on the ambient temperature during testing; (ii) time-of day affected muscular performance only in the neutral condition, (iii) the moderately warm and humid condition blunted the diurnal variation in muscular performance, and (iv) the effect of the ambient temperature was dependent on time of-day. PMID- 15332452 TI - Exposure to light versus way of life: effects on sleep patterns of a teenager- case report. PMID- 15332453 TI - Dental health of Chatham Islanders: an investigation of the oral health of Chatham Islands residents. AB - OBJECTIVES: To examine the self-reported and clinical oral health of Chatham Islanders. DESIGN: A cross-sectional oral health survey. SETTING: The Chatham Islands community. PARTICIPANTS AND METHODS: The investigation involved completion of dental self-report questionnaires by children who attended Chatham Islands schools, and adults who responded to a dental flyer. RESULTS: Eighty seven children and 189 adults took part, all of whom had oral health examinations. Among the children, the prevalence of dental caries in the deciduous dentition was 67.8 percent, and the mean dfs, mean DFS and mean number of missing deciduous teeth due to caries were 5.9 (sd, 6.2), 0.5 (sd, 1.1) and 0.3 (sd, 0.8) respectively. Caries severity was higher in children who were episodic dental attenders, who had irregular brushing habits or whose dental experiences had had a negative impact on their lives. In adults, the prevalence of caries was 81.5 percent. The mean DFS and mean number of missing permanent teeth due to caries were 37.9 (sd, 13.1) and 4.7 (sd, 3.8) respectively. On average, caries severity was higher among infrequent and episodic dental attenders, irregular brushers and those for whom dental problems had had a negative life impact. CONCLUSIONS: While the oral health of Chatham Islands children is comparable with that of children in New Zealand, Chatham Islands adults may be dentally disadvantaged due to limited access to dental resources. The findings have important implications for the planning of dental health services for this remote area, and may provide a useful framework from which further investigations of remote community-dwellers' oral health may be developed. PMID- 15332454 TI - A qualitative study of oral health knowledge and attitudes among staff caring for older people in Dunedin long-term care facilities. AB - Over the next few decades, the number and proportion of older New Zealanders will increase, and many will retain their natural teeth. Many older people in care will be dependent on caregivers for their oral health care. The purpose of this qualitative study was to investigate the dental knowledge and attitudes of staff caring for older people in Dunedin long-term care facilities. In-depth, semi structured interviews with 20 participants from three facilities were recorded on audiotape and subsequently transcribed. The transcripts were analysed to identify themes expressed by the participants. Some of the staff reported poor oral health, and many were irregular dental attenders. Interviewees had a reasonable basic knowledge of dental caries, but were uninformed on many other dental issues, such as the benefits of fluoride and the effects of medication on oral health. All understood that oral health could influence general health. The staff had received very little education about oral health and related issues in their training. There is a need to improve the oral health care knowledge of long-term care facility staff. Incorporation of oral health care into in-service and formal training programmes would be valuable. PMID- 15332455 TI - Continuing professional education in endodontics in New Zealand. AB - OBJECTIVES: To determine the uptake of and wants of New Zealand general dental practitioners for continuing education in endodontics. DESIGN: Mail-out survey questionnaire. MATERIALS AND METHODS: A survey form was sent to general dental practitioners on the Dental Register and data were analysed using SPSS. RESULTS: The response rate was 79 percent. The majority of respondents (92 percent) stated they were either very interested or moderately interested in endodontics, with nearly 34 percent being members of the national endodontic society. Seventy-six percent had attended an endodontic course in the past three years and 85 percent wanted to attend a course. Hands-on courses were the most popular. CONCLUSIONS: Endodontics is a popular discipline in New Zealand, with over 75 percent of respondents having regularly attended refresher courses and wanting to attend further courses. PMID- 15332457 TI - Dental erosion: part 1. Aetiology and prevalence of dental erosion. AB - Non-carious tooth tissue loss due to abrasion, attrition, abfraction and erosion has become a significant problem, occurring in up to 80 percent of children and in up to 43 percent of adults. Dental erosion is now recognised as an important cause of tooth tissue loss in both children and adults. It is caused by the presence of intrinsic or extrinsic acid of non-bacterial origin in the mouth. Intrinsic sources of acid include vomiting, regurgitation, gastro-oesophageal reflux or rumination. Extrinsic sources of acid are most commonly dietary acids. Medications, a patient's lifestyle choices and environment can also increase the risk of dental erosion. In this article we identify the prevalence of dental erosion and its main causative factors, and in Part II we will discuss the management of dental erosion. PMID- 15332456 TI - New Zealand Dental Association practice guidelines on evidence-based dentistry. AB - The importance of evidence in teaching, and to support clinical decisions, is well established in health care, including dentistry. Defending clinical decisions, whether it be with a patient, with a colleague, or at law, increasingly requires reliable data or evidence to support either the treatment plan provided or the treatment given. Recent years have seen an increase in the importance of evidence-based dentistry. This is because of continually changing dental materials and equipment, an increasingly litigious society, an increase in the emphasis of continuing professional development, the information explosion and the consumer movement associated with advances on the Internet. These practice guidelines give a brief review of what evidence-based dentistry is and how to use it. Further details can be found at the New Zealand Dental Association website www.nzda.org.nz. PMID- 15332458 TI - Elective lip piercing and gingival recession: case report. AB - Elective oral piercing as a form of ornamentation or self-expression appears to be gaining popularity in New Zealand and around the world. Several reports of the dangers associated with this practice appear in the dental and medical literature. This report presents a case of rapid, severe gingival recession in a previously periodontally-healthy young female, arising from trauma due to her lip piercing. The aetiology, diagnosis and treatment of her condition are outlined, as well as criteria for the dental community to consider when evaluating patients with facial piercing. The report highlights some of the dangers associated with oral piercing and raises questions concerning potential risk factors for such trauma. PMID- 15332459 TI - The efficacy of casein phosphoprotein-calcium phosphate complex (DC-CP) [Dentacal] as a mouth moistener in patients with severe xerostomia. AB - OBJECTIVES: The purpose of this study was to evaluate casein derivatives coupled with calcium phosphate (CD-CP) (Dentacal, NSI Pty Ltd, Hornsby, Australia) as a mouth moistener in a group of dentate patients with severe xerostomia. DESIGN: The study was a subjective, patient self-evaluation questionnaire on the use of Dentacal compared with the moistening strategies that they usually used. SETTING: The study was hospital and community based. SUBJECTS/MATERIALS, AND METHODS: Thirty eight patients with severe xerostomia were recruited from the larger group of 124 who had taken part in a clinical trial of the anti-caries efficacy of casein derivatives complexed with calcium phosphate (Hay and Thomson, 2002). Each patient used Dentacal for 14 days and the responses to it were compared with the responses to their other mouth moistening strategies. RESULTS: The outcome indicated that Dentacal, when used as an atomised spray in the mouth, provided good moistening and lubrication. CONCLUSIONS: The material could provide benefits in both oral moistening and dental caries prevention in dentate people with xerostomia. PMID- 15332460 TI - Choice of restorative material. PMID- 15332461 TI - The future of education of deaf children: implications of population projections. PMID- 15332462 TI - What verbal protocols reveal about the reading strategies of deaf students: a replication study. AB - Deaf students' reading strategies were identified by investigating these students' self-reported thinking during reading. In an earlier study (Schirmer, 2003), 10 elementary-level students attending a state school for the deaf had constructed meaning, monitored comprehension and activated strategies to improve comprehension, and evaluated comprehension, but had not demonstrated each of the reading strategies within these three overarching activities, all observed in previous studies of hearing skilled readers. Also, the students used a considerably greater variety of reading strategies for constructing meaning than for the other two activities. The replication study used the same procedure. Six elementary-level students attending a site-based public school classroom for deaf students thought aloud after reading each page of a short story. Analysis of these verbal reports indicated the participants performed similarly to those in the first study, thus supporting the reliability of results regarding reading strategies of readers who are deaf. PMID- 15332463 TI - Literacy development in deaf students: case studies in bilingual teaching and learning. AB - A bilingual model has been applied to educating deaf students who are learning American Sign Language (ASL) as their first language and written English as a second. Although Cummins's (1984) theory of second-language learning articulates how learners draw on one language to acquire another, implementing teaching practices based on this theory, particularly with deaf students, is a complex, confusing process. The purposes of the present study were to narrow the gap between theory and practice and to describe the teaching and learning strategies used by the teachers and parents of three elementary school children within a bilingual/bicultural learning environment for deaf students. The findings suggest that strategies such as using ASL as the language of instruction and making translation conceptual rather than literal contribute to literacy learning. Findings further indicate that some inconsistencies persist in applying a bilingual approach with deaf students. PMID- 15332464 TI - Issues in the school-to-work transition of hard of hearing adolescents. AB - Career theorists emphasize the importance of the development of career maturity in adolescents if they are to successfully negotiate the school-to-work transition. Transitions of deaf and hard of hearing adolescents may be especially problematic. The authors examine the implications of current labor market trends for young people, in particular those with hearing loss, and review data on employment outcomes for deaf and hard of hearing people. They discuss the environmental and attitudinal barriers that can influence the career outcomes of this population, consider the impact of hearing loss on adolescents' career maturity, and review the studies on this topic in the literature. The article focuses on the experiences of students with significant hearing loss who are educated in regular classes with the support of itinerant teachers, who communicate orally, and who may be defined as hard of hearing. Recommendations for research and practice are provided. PMID- 15332465 TI - Deaf signers who know Japanese remember words and numbers more effectively than deaf signers who know English. AB - Deaf people have difficulty reading and remembering English script because of its sound-based orthography. Logographs (e.g., kanji, Arabic numerals) should not pose the same challenge because they are based on meaning, not sound. Little research has been conducted to test this theory's validity cross-culturally. The present study was an attempt to do just that. The first of two experiments tested immediate memory spans for word sequences of 20 hearing Irish, 20 prelingually deaf Americans, 20 hearing Japanese, and 20 prelingually deaf Japanese. For English words, deaf participants showed shorter memory spans than hearing participants, but memory spans were similar for deaf and hearing participants for words in kanji, the logographic system for Japanese writing. The second experiment tested memory span for Arabic numerals, with the same participants. Deaf English-readers showed shorter memory spans than their hearing counterparts, but deaf and hearing Japanese performed similarly. PMID- 15332466 TI - Navigating the difficult waters of the No Child Left Behind Act of 2001: what it means for education of the deaf. AB - The author outlines the major elements of the No Child Left Behind Act of 2001 and describes the law's impact on deaf education. The law's stated purpose is to close the achievement gap with accountability, flexibility, and choice, so that no child is left behind. The specific goal of the law is to ensure that all students are 100% proficient in reading, mathematics, and science by 2014. No Child Left Behind has effected sweeping reforms in general education. But with 814 requirements, it has also created great stress in educators throughout the United States. No Child Left Behind poses particular challenges to education of the deaf since policymakers gave no consideration to the needs of deaf children in formulating this law. Clearly, deaf students must be included in school and state accountability systems,but the law leaves many questions unanswered. PMID- 15332467 TI - Organization and use of the mental lexicon by deaf and hearing individuals. AB - Two experiments explored the taxonomic organization of mental lexicons in deaf and hearing college students. Experiment 1 used a single-word association task to examine relations between categories and their members. Results indicated that both groups' lexical knowledge is similar in terms of overall organization, with associations between category names and exemplars stronger for hearing students; only the deaf students showed asymmetrical exemplar-category relations. Experiment 2 used verbal analogies to explore the application of taxonomic knowledge in an academically relevant task. Significant differences between deaf and hearing students were obtained for six types of analogies, although deaf students who were better readers displayed response patterns more like hearing students'. Hearing students' responses reflected their lexical organization; deaf students' did not. These findings implicate the interaction of word knowledge, world knowledge, and literacy skills, emphasizing the need to adapt instructional methods to student knowledge in educational contexts. PMID- 15332468 TI - A rapid ethnography of itinerant teachers of the deaf. AB - A rapid ethnographic study of 10 itinerant teachers in two school districts and 21 other professionals working with the itinerants was conducted. Rapid ethnography starts with the same assumptions about culture as conventional ethnography. However, it is not constrained by the assumption of cultural ignorance on the investigator's part. Thus, it enables better-directed data collection. Interviews with the itinerants and other professionals, direct observation of itinerants at work, and archival data permitted the authors to generate a list of themes reflecting results of other studies which focused on specific skills for itinerant teaching. While knowledge of specific skills cannot be ignored, the study shows that effective itinerants are ones who, through a personality trait, extensive experience, or a specific value system, can generate a positive composite image of their role as itinerants and are then able to interact on the basis of that image. PMID- 15332469 TI - On listening to time passing. PMID- 15332470 TI - Caregiver bereavement outcome: relationship with hospice at home, satisfaction with care, and home death. AB - This study used a randomized controlled trial design to investigate the impact of hospice at home (HAH) on caregiver bereavement outcome. Secondary analyses considered the association between bereavement, place of death, and carers' assessment of support. Ninety-six informal carers of patients referred to HAH were surveyed six weeks post-bereavement about the quality of terminal care. Carers next completed measures of their own bereavement response and general health six months post-bereavement. There was no evidence that HAH had an impact on bereavement outcome. In contrast, perceptions of inadequate terminal support and high symptom severity were associated with worse carer bereavement response. However, it remains unclear whether carers' retrospective ratings constitute an accurate account of symptoms and care. Home deaths were associated with both better bereavement response and better physical health post-bereavement than were inpatient deaths. Further research is needed to investigate the implications of death at home for the carer. PMID- 15332471 TI - Palliative care volunteers: why do they do it? AB - Two studies were conducted to examine people's motives for joining a palliative care volunteer program. To generate a pool of reasons for becoming a palliative care volunteer, previous studies of motivations relevant to palliative care were reviewed and interviews were conducted with 15 palliative care volunteers (Study 1). Combining the literature review and interviews, a total of 22 distinct reasons for volunteering were identified and used to create an Inventory of Motivations for Palliative Care Voluntarism (IMPCV). In Study 2, 113 palliative care volunteers responded to the IMPCV. "To help ease the pain of those living with a life-threatening illness" was rated as the most influential reason for becoming a palliative care volunteer. A principal components factor analysis was conducted on the IMPCV. It was decided that four factors adequately represented the items: Leisure, Personal Gain, Altruism, and Civic Responsibility. PMID- 15332472 TI - Quality end-of-life care in long-term care facilities: service providers' perspective. AB - The goal of this project was to provide guidance on what constitutes quality end of-life care in long-term care (LTC) facilities. Seventy-nine direct care providers from six LTC facilities participated in 12 focus groups. The focus group discussions examined what made the difference between a "good" death and a "bad" death, and what changes in LTC would improve the care of dying residents. Analyses of the focus group data revealed six themes that contribute to quality end-of-life care in LTC facilities: responding to resident needs, creating a homelike environment, supports for families, providing quality care processes, recognizing death as a significant event, and having sufficient institutional resources. These findings challenge policy makers and providers to consider how to normalize life and death in LTC facilities. PMID- 15332473 TI - Family responses to declining intake and weight loss in a terminally ill relative. Part 1: fighting back. AB - We recently completed a grounded theory study examining nutritional care experiences in advanced cancer from the perspective of patients (n=13), families (n=23), and health care providers (n=11) (1). That work generated important information about adult family members' perceptions and behaviour regarding the nutritional care their terminally ill adult relative received while hospitalized on an inpatient palliative care unit. An overview of the inductively derived model that emerged from that work has been reported elsewhere (2). This article provides a more detailed description of one of the major sub-processes of the model regarding family member responses to declining oral intake and weight loss in a terminally ill relative-the sub-process of "fighting back: it's best to eat." The strategies family members use when fighting back, and the consequences of these strategies for patients, family members, and health care providers are reported. Implications for practice and research are provided. PMID- 15332474 TI - Thrombosis and anticoagulation in palliative care: an evolving clinical challenge. AB - There is a clear recognition of the relationship between thrombosis and neoplastic diseases. In recent years, there have been a few reports dealing with some of the features of clinical progression and management of venous thromboembolism (VTE) in palliative care patients. In this report, we present four cases that describe some of the diverse aspects of this clinically challenging area to illustrate the symptom and quality of life issues for patients, and the management challenge for palliative care clinicians. Perhaps the most important issue in the palliative care setting is the need to balance the inconvenience of the various diagnostic and therapeutic options, particularly in settings where progression has resulted in a deteriorated physical and cognitive condition. We need to recognize that VTE does cause significant problems for some of our patients. We will always need to balance issues of life expectancy, comfort and quality of life, and risk and benefits of anticoagulation, and align these problems with the individual patient, and the family dynamics and desires we encounter. PMID- 15332475 TI - What are the key components of quality perinatal and pediatric end-of-life care? A literature review. PMID- 15332476 TI - Family perceptions of clinicians' outstanding practices in end-of-life care. PMID- 15332477 TI - High-dose levomepromazine (methotrimeprazine) to control nausea in carcinoid syndrome. PMID- 15332478 TI - Suicidal intent in the palliative care setting. PMID- 15332479 TI - Malaria specific-IgG, inter-pregnancy intervals, birth weights and body mass index in cases of asymptomatic malaria parasitaemia. AB - BACKGROUND & OBJECTIVES: Considerations of both inter-pregnancy intervals and malaria parasitaemia may help in understanding some aspects of susceptibility and pregnancy outcomes in malaria endemic areas. METHODS: Pregnant women with asymptomatic malaria parasitaemia were recruited and divided into groups based on their inter-pregnancy intervals and malaria specific-IgG, body mass index, and birth weights were studied in the groups. RESULTS: The results showed that the P. falciparum specific-IgG concentration (f=3.52, p<0.02), malaria parasites density (f=6.44, p<0.001) and birth weights (f=7.36, p<0.001) were significantly different amongst the groups with varying inter-pregnancy intervals. In addition, different levels of associations between variables such as 'inter-pregnancy intervals vs P. falciparum specific-IgG concentration' (r = 0.23, p<0.05); 'malaria parasites density vs birth weight' (r = -0.84, p < 0.01) was observed. INTERPRETATIONS & CONCLUSION: This study suggests that inter-pregnancy intervals could be one of the factors influencing dynamic serum concentrations of P. falciparum specific-IgG while malaria parasitaemia could be one of the factors affecting birth weights. Hence, observance of inter-pregnancy intervals has its own implications in malaria endemic areas. PMID- 15332480 TI - Epidemiological and entomological aspects of malaria in forest-fringed villages of Sonitpur district, Assam. AB - BACKGROUND & OBJECTIVES: Detailed epidemiological and entomological studies were undertaken in forest-fringed villages and a Tea Estate in Sonitpur, Assam to assess the malaria situation. METHODS: Door-to-door active surveillance was carried out to collect blood samples. Thick and thin blood smears stained with Giemsa were used for malaria parasite detection. Mosquito collections were made using CDC miniature light-traps and hand catch methods from dusk-to-dawn. RESULTS: 48% SPR, 49.1 Pf% was recorded from the study villages. Children between 10 and 14 years were most sufferers. Per trap night density of mosquitoes in human dwellings was 204.3 and in cattlesheds--908.7, An. minimus accounted for 20.7% of total malaria vectors. INTERPRETATION & CONCLUSION: The results showed high malaria risk in the study villages. High vector density with high parity rate, poor socio-economic conditions, lack of awareness, poor sanitation and congenial atmosphere for mosquito proliferation are aggravating the malaria situation more complex in the study area. PMID- 15332481 TI - Evaluation of pirimiphos-methyl (50% EC) against the immatures of Anopheles stephensi/An. culicifacies (malaria vectors) and Culex quinquefasciatus (vector of bancroftian filariasis). AB - BACKGROUND & OBJECTIVES: In India, temephos and fenthion are used as larvicides in fresh and polluted waters. Since use of same insecticide may precipitate resistant, as an alternative bioefficacy of pirimiphos-methyl--an organophosphorus insecticide was evaluated against immatures of Anopheles and Culex species in different breeding habitats in District Ghaziabad (U.P.) and Goa. METHODS: Laboratory bioassays were carried out using standard WHO procedure. LC50 and LC90 were calculated using Probit analysis. Abbott's formula was used to calculate per cent corrected mortality in laboratory exposures while, Mulla's formula was applied for calculating per cent reduction in mosquito immatures under field conditions. RESULTS: Pirimiphos-methyl was found to be most effective against larvae of An. stephensi followed by An. culicifacies and Cx. quinquefasciatus. The LC50 and LC90 values against three species were 0.023, 0.032 and 0.04 ppm; and 0.045, 0.057 and 0.114 ppm respectively. In field, pirimiphos-methyl @ 200 g a.i./ha against Anopheles species and @ 300 g a.i./ha against Culex species can produce > 80% reduction in density of immatures up to one week in different habitats. Pirimiphos-methyl < 0.25 ppm is not toxic to fish. INTERPRETATION & CONCLUSION: Pirimiphos-methyl is more effective against anophelines in clean water than culicines in polluted water. This larvicide should not be used at a concentration above 0.25 ppm in habitats harbouring the larvivorous fish. PMID- 15332482 TI - Altitudinal distribution of mosquitoes in mountainous area of Garhwal region: Part-I. AB - BACKGROUND & OBJECTIVES: Mosquito fauna diversity in mountainous areas of Garhwal region was studied during November 2000 to October 2002 to correlate the altitudinal vegetation and distribution of mosquitoes. METHODS: Adult mosquitoes and mosquito immatures were collected using WHO methods and identified using standard keys and catalogues. Altitude of mosquito habitat was measured using portable altimeter and also by GPS. RESULTS: Altogether 34 species in five genera -Aedes, Anopheles, Armigeres, Culex and Uranotaenia were encountered in the present study in the altitude range of 300 to 2000 m. Majority of the mosquitoes were found in between 300 to 900 m altitude except Culex vagus and Anopheles maculatus, which were found throughout the range. INTERPRETATION & CONCLUSION: The mosquitoes were categorised into six groups based on their altitudinal distribution. The areas at lowest elevation were having the greatest number of species but not the corresponding greater number of specimens in the present study. PMID- 15332483 TI - Assessment of knowledge about malaria among patients reported with fever: a hospital-based study. AB - BACKGROUND & OBJECTIVES: Knowledge about malaria, attitude and health seeking behaviour of fever patients attending Medicine Out Patient Department at Safdarjang Hospital, New Delhi were studied from June to August 2003. METHODS: Cross-sectional survey comprising 200 fever cases was done with prestructured and pretested questionnaire. Data on sociodemographic profile, history of fever, health seeking behaviour, etc were recorded. RESULTS: About 83% of fever cases did not approach the doctor even after three days of onset of fever symptom, 25.5% tried self medication and 20.5% approached chemists for treatment. Knowledge about causes and prevention of malaria was found to be inadequate in the study subjects. INTERPRETATION & CONCLUSION: Knowledge about malaria is poor even in persons residing in urban localities and proper health education is required for successful control of malaria. Information, education and communication activities are indicated to create awareness among the community. PMID- 15332484 TI - Outdoor resting preference of Culex tritaeniorhynchus, the vector of Japanese encephalitis in Warangal and Karim Nagar districts, Andhra Pradesh. AB - BACKGROUND & OBJECTIVES: Entomological investigations were carried out in Warangal and Karim Nagar districts of Andhra Pradesh during viral encephalitis outbreak in July 2003. METHODS: Adult mosquito collections in outdoor were done using sweep cage method and in indoor with the help of aspirator tube and flash light. Larval collections were done by dipping method. JE virus positivity was tested by IgM capture ELISA test. RESULTS: Outdoor collections revealed presence of Culex tritaeniorhynchus, Cx. bitaeniorhynchus and Cx. gelidus and in indoor collections--Cx. quinquefasciatus, Cx. tritaeniorhynchus, An. vagus and An. subpictus. In the outdoor collections Cx. tritaeniorhynchus was predominant (96.3% of total collection). Three samples out of 55 serum samples from human cases and five from contacts showed the presence of antibodies against JE virus. INTERPRETATION & CONCLUSION: Cx. tritaeniorhynchus, a known vector of JE is predominant in outdoors and playing a main role in JE transmission in this area. Vector control aimed at the outdoor resting population might limit virus circulation in the mosquito vertebrate host cycle and prevent human infection. PMID- 15332485 TI - Variations in ornamentation of wings and palpi of Anopheles (Cellia) subpictus Grassi collected from northwest India. PMID- 15332486 TI - A report on the susceptibility status of Anopheles minimus (Theobald) against DDT and deltamethrin in three districts of Assam. PMID- 15332487 TI - Natural killer cells in HIV-1 infection: role of NK cell-mediated non-cytolytic mechanisms in pathogenesis of HIV-1 infection. AB - Natural killer (NK) cells exhibit both cytolytic and non-cytolytic effector functions against HIV-infected targets. Their precise role in immunopathogenesis of HIV-1 infection is yet to be fully understood. This review addresses the non cytolytic functions exhibited by NK cells, their potential role in pathogenesis of HIV-1 infection and the effect of HIV-1 viremia on NK cell functions. Activated NK cells are capable of secreting CC-chemokines and suppressing HIV-1 replication in a non-cytolytic fashion. However, HIV-1 viremia suppresses the ability of NK cells to secrete CC-chemokines. Suppression of HIV-1 viremia by highly active antiretroviral therapy (HAART) restores the ability of NK cells to secrete CC-chemokines and suppress endogenous HIV-1 replication by non-cytolytic mechanisms. Better understanding of the mechanisms involved in HIV-1-NK cell interactions would be helpful in delineating novel therapeutic strategics against HIV-1. PMID- 15332488 TI - Antituberculosis drug-induced hepatitis: risk factors, prevention and management. AB - Apart from infectious or viral hepatitis, other most common non-infectious causes of hepatitis are alcohol, cholestatic, drugs and toxic materials. The most common mode that leads to liver injuries is antituberculosis drug-induced hepatitis. The severity of drug-induced liver injury varies from minor nonspecific changes in hepatic structure to fulminant hepatic failure, cirrhosis and liver cancer. Patients receiving antitubercular drug frequently develop acute or chronic hepatitis. The time required for the metabolites to reach hepatotoxic levels is much earlier with isoniazid plus rifampicin treatment than isoniazid alone and this has been shown to be synergistic rather than additive. Antituberculosis drug (ATT)-inducible cytochrome P-4502E1 (CYP2E1) is constitutively expressed in the liver. Recent studies show that polymorphism of the N-acetyltransferase 2 (NAT2) genes and glutathione-S-transferase (GST) are the major susceptibility risk factors for ATT-induced hepatitis. The hepatic NAT and GST are involved in the metabolism of several carcinogenic arylamines and drugs. The NAT2 enzyme has a genetic polymorphism in human. N-acetyltransferase 2 genes (NAT2) have been identified to be responsible for genetic polymorphism of slow and rapid acetylation in humans. Slow acetylators of NAT2 prove to develop more severe hepatotoxicity than rapid acetylators making it a significant risk factor. Deficiency of GST activity, because of homozygous null mutations at GSTM1 and GSTT1 loci, may modulate susceptibility to drug and xenobiotic-induced hepatotoxicity. Polymorphisms at GSTM1, GSTT1 and NAT2 loci had been linked to various forms of liver injury, including hepatocellular carcinoma. PMID- 15332489 TI - Internet--implications for the future of phytopharmacological research. AB - Modern information technologies and world wide communications through the Internet play a significant role in medicinal plant research across the globe. The phenomenal growth in Internet usage is largely due to the success of World Wide Web. Various useful websites and databases on phytopharmacology are already in the "Net" and many more are being added constantly. The future of phytopharmacological research is handling the existing information in proper way. In this review of the Internet, compilation of important websites is expected to stimulate, instruct and update academicians and researchers involved in phytopharmacological research. PMID- 15332490 TI - Bioremediation of paper and pulp mill effluents. AB - Pulp and paper mill effluents pollute water, air and soil, causing a major threat to the environment. Several methods have been attempted by various researchers throughout the world for the removal of colour from pulp and paper mill effluents. The biological colour removal process uses several classes of microorganisms--bacteria, algae and fungi--to degrade the polymeric lignin derived chromophoric material. White rot fungi such as Phanerochaete chrysosporium, Corius versicolor, Trametes versicolor etc., are efficient in decolourizing paper and pulp mill effluents. Gliocladium virens, a saprophytic soil fungus decolourised paper and pulp mill effluents by 42% due to the production of hemicellulase, lignin peroxidase, manganese peroxidase and laccase. PMID- 15332491 TI - Localization of bypass-induced changes in flow in coronary artery models. AB - Right coronary artery bypass restores blood flow through heart tissues. This also induces changes in flow leading to its failure. By this work the sites which are prone to such changes are localized. The bypass models are developed from transparent silicon rubber of elastic properties similar to arterial tissues. Flow visualization is carried out by photoelasticity technique by using dilute solution of vanadium pentoxide. This analysis carried out under pulsatile flow conditions shows that the proximal stenotic region continues to contribute to the alteration in flow in the hood region of the bypass. Thus making its proximal and distal regions prone to flow-induced changes, which may lead to its blockage over the long duration. PMID- 15332492 TI - Enhancement of metastatic potential of mouse B16-melanoma cells to lung after treatment with gangliosides of B-16-melanoma cells of higher metastatic potential to lung. AB - Mouse B16LuF1 melanoma cells of lower metastatic potential to lung were treated in vitro with same concentration (50 microM) of gangliosides isolated from B16LuF5, B16LuF9 or B16LuF10 cells with higher metastatic potential to lung (LuF1< LuF5< LuF9< LuF10) and injected to groups of normal mice through tail vein. The number of metastatic tumor nodules formed in lung increased in mice receiving B16LuF5, B16LuF9 and B16LuF10-ganglioside-treated B16LuF1 cells compared to mice receiving B16LuF1 cells without any ganglioside treatment. Metastatic potential of B16LuF1 cells gradually increased after treatment with gangliosides of B 16-melanoma cells of increasing metastatic potential to lung. The six major gangliosides isolated from B16LuF10 cells corresponded with standard gangliosides GT1b, GD1b, GD1a, GM1, GM2 and GM3 respectively on TLC analysis. When B16LuF1 cells were treated in vitro with each of these six individual gangliosides and injected to groups of normal mice through tail vein the number of tumor nodules formed in lung varied. The four groups of mice receiving B16LuF1 cells treated with each of four gangliosides corresponding to GT1b, GD1b, GD1a or GM1 produced lung metastasis comparable to that of untreated control group. Only remaining two gangliosides which corresponded with standard gangliosides GM2 and GM3 increased metastatic potential of B16LuF1 cells. Thus, these results indicated that gangliosides GM2 and GM3 of B16-melanoma cells are definitely associated with metastatic potential of these tumor cells. PMID- 15332493 TI - Effect of alcohol on neurons of iso-cortex--a histomorphometric study. AB - Effect of chronic intake of alcohol and its subsequent withdrawal was studied in albino mice on the layers of neurons of the iso-cortex. Neuronal density per mm2 of section in different layers of iso-cortex was counted and compared in 3 groups of animals (control, ethanol fed and withdrawal). Qualitative changes on nissl granules of neurons and myelinated fibres were also studied. Mice fed with 10% ethanol v/v ad libitum for 6 months showed loss of nissl granules and nucleolus and discontinuity of nuclear membrane. Quantitatively, significant reduction in neuronal density (P<0.001) was observed in layers II+III IV and V neurons of iso cortex. Withdrawal of ethanol for 2 months showed continued reduction of counts of neuronal density in layers II+III and V only whereas reversal of count was found significantly (P<0.001) in layer IV of iso-cortex. Qualitatively, only few neurons showed prominent nissl granules after withdrawal of ethanol. More afferent synaptic connection in layer IV may be suggested as probable factor helping relative replenishment of neuronal count after withdrawal of alcohol. PMID- 15332494 TI - FK506 as effective adjunct to L-dopa in reserpine-induced catalepsy in rats. AB - Reserpine-induced catalepsy is a widely accepted animal model of Parkinson's disease. In the present study reserpine (2.5 mg/kg, ip) 20 hr and alpha-mehyl para-tyrosine (AMPT; 200 mg/kg, ip), one hour before the experiment induced significant catalepsy in rats as assessed by bar test. There was a significant increase in the time spent on the bar in bar test as compared to the control untreated rats. L-dopa (100 mg/kg, ip) and carbidopa (10 mg/kg, ip) combination, a conventional therapy was less effective in reversing reserpine-induced catalepsy. Pretreatment with FK506, a neuroprotectant (0.5-2 mg/kg, po) not only dose dependently reduced the catalepsy in reserpine-treated rats but a lower dose (1 mg/kg) potentiated the motor stimulant actions of sub threshold dose of L-dopa (100 mg/kg, ip) and carbidopa (10 mg/kg, ip) combination. Anticataleptic effect of FK506 was blocked dose dependently by specific D2 receptor blocker sulpiride (25-100 mg/kg, ip). In conclusion, the findings of the present study suggest that FK506 has an indirect modulatory action on the dopamine D2 receptors. FK506 being a neuroprotectant, could be used as an effective adjunct to L-dopa for the treatment of neuroleptic-induced extrapyramidal side effects. PMID- 15332495 TI - Evaluation of cognitive function of fluoxetine, sertraline and tianeptine in isolation and chronic unpredictable mild stress-induced depressive Wistar rats. AB - Depressive illness is generally associated with cognitive impairment. Serotonergic selective antidepressant drugs, fluoxetine (FLX), sertraline (SER) and tianeptine (TIA), are claimed to have less or no effect on cholinergic system, the key system involved in memory. In the present study, these drugs were evaluated for their influence on cognitive behavior in both depressive and non depressive animals. Depression was induced by two models, (i) 60 days social isolation of litter; and ii) by applying chronic unpredictable mild stress for 21 days. Depression in the rats was confirmed by behavioral despair test. Transfer latency on elevated plus maze and inflexion ratio in passive avoidance step through behavior were employed to assess learning and memory. The results indicated that administration of fluoxetine; sertraline and tianeptine attenuated the cognitive deficits observed in depressive rats. In non-depressive rats these drugs produced retention deficit, which was found to be parameter and model dependent. Data suggested that, FLX and SER (SSRI's) effectively attenuated the isolation-induced depression and cognitive deficit, whereas TIA (SSRE) produced better effect in stress-induced depressive conditions. It was concluded that behavioral profiles of fluoxetine, sertraline and tianeptine on cognition were model and parameter dependent. PMID- 15332496 TI - Suicidal oxidative stress induced by certain antioxidants. AB - Well known antioxidants-coumarins (7,8-dihydroxy-4-methyl coumarin-DHMC and 7,8 diacetoxy-4-methyl coumarin-DAMC) and flavonoids (quercetin-Q and quercetin penta acetate-QPA) were investigated for their pro-oxidant effects in two human tumor cell lines. The breast carcinoma cell line (MDA-MB-468) was found to be more sensitive to treatment by the drugs-DAMC, Q and QPA at 10 microM than the glioma cell line (U-87MG), while DHMC was non toxic in both cell lines at this concentration. In MDA-MB-468 distinct growth inhibition was observed by 48 hr post treatment. Paradoxically, an increase in the formazan production was revealed by MTT assay at this time indicating an increase in the production of free radicals. An increase in the levels of reactive oxygen species (ROS) was also confirmed by DCFH-DA assay. In cells treated with DAMC, Q and QPA an increase in the percentage of cells with the hypodiploid DNA content was suggestive of apoptotic cell death. Taken together, these results suggest that an increase in oxidative stress caused by the pro-oxidant action of these drugs is responsible for cell death. PMID- 15332497 TI - Central inhibitory effect of Moringa oleifera root extract: possible role of neurotransmitters. AB - Effect of chronic treatment of standardized aqueous extract of Moringa oleifera (MO) root (100, 200, 300, 350, 400, 450 mg/kg; po) on penicillin (PCN) induced convulsion, locomotor behaviour, brain serotonin (5-HTT), dopamine (DA) and norepinephrine (NE) level was studied in Holtzman strain adult albino rats. The result revealed that pretreatment with MO inhibited PCN-induced seizure and markedly reduced locomotor activity. Chronic treatment with MO significantly increased the 5-HT and decreased the DA level in cerebral cortex (CC), midbrain (MB), caudate nucleus (CN) and cerebellum (CB). NE level was significantly decreased in CC but no appreciable change was observed in MB, CB and CN. Thus the central inhibitory effect of MO is discussed in the light of the disturbed balance between 5-HT, DA and NE. PMID- 15332498 TI - Effect of Aegle marmelos Correa. (Bael) fruit extract on tissue antioxidants in streptozotocin diabetic rats. AB - A study was undertaken to evaluate the anti-lipid peroxidative activity of an aqueous extract of A. marmelos fruits (AMFEt) in streptozotocin diabetic rats in heart and pancreas. Oral administration of AMFEt for 30 days (125 and 250 mg kg( 1) body weight twice daily) produced a significant decrease in the elevated levels of peroxidation products, viz. thiobarbituric acid reactive substances and hydroperoxides in the tissues of diabetic rats. The depressed activities of superoxide dismutase, catalase and glutathione peroxidase and lowered glutathione content in the heart and pancreas of diabetic rats were found to increase on treatment with AMFEt. AMFEt at a dose of 250 mg kg(-1) was more effective than glibenclamide (300 microg kg(-1)) and both reversed all the values significantly. Thus AMFEt exhibits anti-oxidative activity in streptozotocin diabetic rats. PMID- 15332499 TI - Effect of housing rats within a pyramid on stress parameters. AB - The Giza pyramids of Egypt have been the subject of much research. Pyramid models with the same base to height ratio as of the Great Pyramid of Giza, when aligned on a true north-south axis, are believed to generate, transform and transmit energy. Research done with such pyramid models has shown that they induced greater relaxation in human subjects, promoted better wound healing in rats and afforded protection against stress-induced neurodegnerative changes in mice. The present study was done to assess the effects of housing Wistar rats within the pyramid on the status of oxidative damage and antioxidant defense in their erythrocytes and cortisol levels in their plasma. Rats were housed in cages under standard laboratory conditions. Cages were left in the open (normal control), under a wooden pyramid model (experimental rats) or in a cubical box of comparable dimensions (6 hr/day for 14 days). Erythrocyte malondialdehyde and plasma cortisol levels were significantly decreased in rats kept within the pyramid as compared to the normal control and those within the square box. Erythrocyte reduced glutathione levels, erythrocyte glutathione peroxidase and superoxide dismutase activities were significantly increased in the rats kept in the pyramid as compared to the other two groups. There was no significant difference in any of the parameters between the normal control and rats kept in the square box. The results showed that exposure of adult female Wistar rats to pyramid environment reduces stress oxidative stress and increases antioxidant defense in them. PMID- 15332500 TI - Effect of a polyherbal formulation, Ambrex, on butylated hydroxy toluene (BHT) induced toxicity in rats. AB - Effect of polyherbal formulation Ambrex was evaluated in butylated hydroxytoluene (BHT) induced toxicity of lungs and liver in rats. Toxicity was produced by administering BHT (500 mg/kg/day) for 3 days. Lung damage was evidenced by elevated levels of broncho alveolar lavage fluid (BAL) parameters such as protein, lactate, lactate dehydrogenase (LDH), alkaline phosphatase (ALP), acid phosphatase (ACP) and glucose-6-phosphate dehydrogenase (G6PDH). Liver damage was proved by elevated levels of serum protein and markers such as LDH, ALP, aspartate amino transferase (AST), alanine amino transferase (ALT), decreased level of lipid peroxides (LPO) in serum and glutathione (GSH) in liver. Administration of aqueous suspension of Ambrex (50 mg/kg orally) retained these elevated levels of BAL-protein, lactate, LDH, ALP, ACP, G6PDH and serum-protein, LDH, ALP, AST and ALT at near normal values. Decreased level of liver GSH was retained at near normalcy in Ambrex pretreated BHT-administered animals. There was no change in liver LPO in all the four groups. PMID- 15332501 TI - Effect of arsenic on cell growth of the cellular slime mould, Dictyostelium discoideum. AB - The growing D. discoideum cells were killed in a dose-dependent manner when exposed to 100 and 140 ppm of arsenic (As2O3) at mid-log phase for 20 min. Reduced plaque sizes and changed cell and colony morphologies were observed in the treated cells. Endocytotic functions (both phagocytosis and pinocytosis) were also inhibited in the treated cells. Arsenic treated cell showed a lower DNA and protein synthetic activities. These findings are discussed in relation to known mechanism of action of the heavy metal on growth-related cellular functions. PMID- 15332502 TI - Oxygen uptake and filtration rate as animal health biomarker in Lamellidens marginalis (Lamarck). AB - The freshwater bivalve, L. marginalis was experimentally exposed to 10 and 30 ppm concentrations of CdCl2 to examine filtration rate, oxygen uptake and glycogen level of liver and gills for health assessment for their reuse in the reclamation of cadmium intoxicated environments. In situ experiment was also performed for better appraisal of the filtration rate in the lake. Oxygen uptake in the treated group exceeded that of control by 15-22% during the early 24 hr after cadmium exposure, but followed an essential decline (23-30%) thereafter. The reduction of filtration rate ranged from 12-62% in laboratory to 83-85% in field trials. At the tissue level, glycogen content was reduced by 61-72% in liver and 52-63% in gill. In both tissues, glycogen content was inversely proportional to the cadmium contents of the animal. Critical appraisal of data suggests that the threshold values of cadmium in gill and liver were 50-80 microg/g dw for oxygen uptake and 50-60 microg/g for filtration rate because of marked reduction of these parameters beyond the values of cadmium. It is concluded that filtration rate, oxygen uptake of the freshwater bivalve, L. marginalis can be used as biomarker for animal health assessment and for possible reuse of the stock animals. PMID- 15332503 TI - In vitro control of fasciation in proliferating nucellar embryos of Mangifera indica L. var totapari red small for cloning. AB - Nucellar tissue contained in ovular halves of young fruits of Mangifera indica L. totapari red small, a dwarfing rootstock, differentiated fasciated embryonal structures in presence of 6-benzylaminopurine [BAP(0.15 mg l(-1))], 6-(gamma gamma-dimethylallylamino) purine [2iP(0.15 mg l(-1))] and indole-3-acetic acid [(IAA(0.5 mg l(-1))] incorporated in the semisolid medium during 50-60 days. Due to embryonal fasciation, hardly 2-3 well-formed embryos could be obtained per culture of proliferating embryos. Of the 3 ethylene inhibitors [L-alpha-(2 aminoethoxyvinyl)-glycine-HCl (AVG), AgNO3 and salicylic acid (SA)] used, embryonal fasciation and necrosis of intervening tissue was completely controlled by 3-4 subcultures of fasciated mass of embryos under the influence of AVG (0.05 mg l(-1)) in presence of adenine sulphate [AdS (50 mg l(-1))] incorporated in the same medium. Almost synchronized development of isolated embryos, measuring ca 2 cm in length, was observed in a different medium used in liquid stationary state and supplemented, particularly with stress-producing substances [abscisic acid (ABA, 0.01 mg l(-1)); and polyethylene glycol (PEG, 100 mg l(-1))] besides certain other modifications. About 34% convertibility of processed embryos was obtained during a period of 90 days. The plantlets had well-developed roots along with laterals which were longer than leafy shoots. In vitro raised plants survived ex vitro for about 2 months. PMID- 15332504 TI - Modulatory influence of Brassica compestris Linn var sarson on phase-II carcinogen metabolizing enzymes and glutathione levels in mice. AB - The present study reports the modulatory influence of 95% ethanolic extract from the seeds of B. compestris on the activity of phase-II enzymes such as glutathione S-transferase (GST), DT-diaphorase (DTD) and reduced glutathione (GSH) level in the skin, lung, kidney and forestomach of the mouse. Oral treatment with the seed extract at 800 mg/kg body wt. for 15 days significantly elevated GST in lung and forestomach and DT-diaphorase in forestomach and skin and GSH level in lung, kidney forestomach and skin. The lower dose 400 mg/kg body wt was effective only in inducing GST and DT-diaphorase activity in forestomach and reduced glutathione level in lung. The findings suggest that B. compestris seed extract may block or suppress the events associated with chemical carcinogenesis at least in part, by inducing metabolic detoxification of the carcinogen. PMID- 15332505 TI - Influence of acidic beverage (Coca-Cola) on pharmacokinetics of ibuprofen in healthy rabbits. AB - The study was aimed at determining the effect of Coca-Cola on the pharmacokinetics of ibuprofen in rabbits. In a cross-over study, ibuprofen was given orally in a dose of 56 mg/kg, prepared as 0.5% suspension in carboxymethyl cellulose (CMC) and blood samples (1 ml) were drawn at different time intervals from 0-12 hr. After a washout period of 7 days, Coca-Cola in a dose of (5 ml/kg) was administered along with ibuprofen (56 mg/kg) and blood samples were drawn from 0-12 hr. To these rabbits, 5 ml/kg Coca-Cola was administered once daily for another 7 days. On 8th day, Coca-Cola (5 ml/kg) along with ibuprofen (56 mg/kg), prepared as a suspension was administered and blood samples (1 ml each) were drawn at similar time intervals. Plasma was separated and assayed for ibuprofen by HPLC technique and various pharmacokinetic parameters were calculated. The Cmax and AUC0-alpha of ibuprofen were significantly increased after single and multiple doses of Coca-Cola, thereby indicating increased extent of absorption of ibuprofen. The results warrant the reduction of ibuprofen daily dosage, frequency when administered with Coca-Cola. PMID- 15332506 TI - Chemopreventive action of Phyllanthus urinaria Linn on DMBA-induced skin carcinogenesis in mice. AB - The inhibition of tumor incidence by hydro-alcoholic extract of the whole plant of P. urinaria was evaluated in 6-7 weeks old female albino mice on two-stage process of skin carcinogenesis induced by a single application of 7,12 dimethylbenz(a)anthracene (50 microg/50 microl of acetone), and 2 weeks later, promoted by repeated application of croton oil (1% in acetone/three times a week) till the end of the experiment (15 weeks). Topical application of the extract at a dose of 5 mg/kg body weight/day for 15 weeks at the peri-initiational stage (i.e., 7 days before and 7 days after DMBA application), promotional stage (i.e., from the time of croton oil application) and both peri and post-initiational stages (i.e., 7 days prior to DMBA application and continued till the end of the experiment) on the shaven backs of the mice recorded a significant reduction in tumor incidence to 50, 33.3 and 16.7% respectively in comparison to the control (i.e., the mice treated with DMBA and croton oil only) where tumor incidence was found to be 81.8%. The average number of papillomas per mouse was also significantly reduced. The results suggest a possible chemopreventive property of P. urinaria against DMBA-induced skin papillomagenesis in mice. PMID- 15332507 TI - Anticonvulsant potential of holy basil, Ocimum sanctum Linn., and its cultures. AB - Callus cultures from stem of O. sanctum were induced on slightly modified Murashige and Skoog's (MS) medium and supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, 1-2 ppm) and kinetin (kn, 1 ppm). Different extractives of stem, leaf and stem callus of O. sanctum were tested for anticonvulsant activity against standard drug phenytoin using maximal electroshock (MES) model. Ethanol and chloroform extractives of stem, leaf and stem calli were effective in preventing tonic convulsions induced by transcorneal electroshock. PMID- 15332508 TI - Changes in lipid peroxidation and free radical scavengers in kidney of hypothyroid and hyperthyroid rats. AB - Kidney weight was significantly decreased in hypothyroidism (induced by Na131I administration) and increased in hyperthyroidism (induced by thyroxine treatment) as compared to control in female Wistar rats. The tissue lipid peroxidation level remained unchanged in hyperthyroid rats but significantly increased in hypothyroid rats. Superoxide dismutase was decreased in both experimental groups but more so in hyperthyroid rats. Catalase was reduced significantly in hyperthyroid rats but remained unaffected in hypothyroid rats. Tissue glutathione peroxidase (GPx) activity was increased while reduced glutathione levels remained unaltered in both hypothyroid and hyperthyroid rats. Plasma GPx activity was significantly low in both the hypothyroid and hyperthyroid rats. The results suggest alterations in the oxidative stress in hypothyroid and hyperthyroid rat kidneys with concomitant changes of free radical scavengers. PMID- 15332509 TI - In vitro lethal efficacy of leaf extract of Cannabis sativa Linn on the larvae of Chironomous samoensis Edward: an insect of public health concern. AB - Leaf extract of C. sativa causes paralysis leading to death in larvae of C. samoensis. The extract brought a drastic change in the morphology of sensilla trichoidea and the general body cuticle. The larvae exposed to the leaf extract also showed a significant reduction in the concentration of Mg and Fe, while Mn showed only slight average increase. Since the sensilla trichoidea has nerve connection, it is expected that the toxic principle of the leaf extract has affected the central nervous system. The significant reduction of the level of Fe indicates that the extract could cause the reduction in oxygen binding capacity of the haemolymph, thereby acting as a respiratory poison in addition to its known role as a neurotoxic substance. PMID- 15332510 TI - Dietary requirement of tryptophan for growth and survival of the Indian major carp, Cirrhinus mrigala (Hamilton-Buchanan) fry. AB - The experimental diets (40% protein) contained graded levels of tryptophan (0.60, 0.75, 1.00, 1.25, 1.50, 1.75%) of dietary protein. Each test diet was fed to triplicate groups of mrigal fry twice daily at 10% of the total biomass for 8 weeks. The dietary tryptophan requirement was estimated by plotting weight gain against dietary levels of tryptophan using two separate regression equations, the point of intersection of two equations was taken as optimum level, which occurred at 0.48% of the diet (1.20% of dietary protein). PMID- 15332511 TI - Occurrence and cross infectivity of granulovirus of field bean pod borer Adisura atkinsoni Moore. AB - A granulovirus (GV) was isolated from the field-bean pod borer, Adisura atkinsoni. Electron microscopic observation showed capsule or granular shaped occlusion bodies. The virus was highly virulent against second instar larvae when tested at 1 x 10(6) occlusions/larva through food surface (pod/seed) contamination technique. The incubation period ranged from 6-10 days in the case of second instar larvae. In contrast to green coloured healthy larvae. GV infected A. atkinsoni became brownish/pale white in colour mostly due to accumulation of large number of occlusion bodies. Study on the cross infectivity of A. atkinsoni GV to gram caterpillar, Helicoverpa armigera revealed the high susceptibility of H. armigera to A. atkinsoni GV, thereby widening the scope of controlling both the species on the same cropping system. This is the first record of GV from A. atkinsoni from India. PMID- 15332512 TI - In vitro micropropagation of Baliospermum montanum (Willd.) Muell-Arg--a medicinal plant. AB - Micropropagation of B. montanum was achieved on Murashige and Skoog's (MS) medium augmented with BAP using nodal segments. Maximum number of shoots (3.4 +/- 0.25) were found in MS medium fortified with BAP (3.10 microM). In vitro raised shoots were rooted on half strength MS medium augmented with various concentrations and combination of auxins viz.. IAA, IBA and NAA. Maximum number of roots were observed on half strength MS medium fortified with IBA (9.84 microM) combined with NAA (5.37 microM). PMID- 15332513 TI - Yeast made somewhat human to produce human proteins. PMID- 15332514 TI - New insights into DNA transcription. PMID- 15332515 TI - Rolling in dough. Despite the controversy surrounding CEO pay, healthcare leaders continue to enjoy generous compensation, with insurers leading the pack. AB - Despite the controversy about pay for CEOs, leaders continue to enjoy highly lucrative compensation. Modem Healthcares second annual report on CEO pay shows healthcare chieftains fared nicely. Acute-care hospital CEOs, such as HCA's Jack Bovender Jr., left, saw their median compensation rise 12.50% PMID- 15332516 TI - Dems hang hat on healthcare. Coverage for poor, uninsured in spotlight at DNC. PMID- 15332517 TI - Rules and regulations. CMS addresses drug prices, payment rates. PMID- 15332518 TI - Bad debt rising. HCA's expenses down; others not so lucky. PMID- 15332519 TI - Error measure draws critics. PMID- 15332520 TI - 'Paving the way. Judge brings Pa. closer to capping Blues' surpluses. PMID- 15332521 TI - Lone holdout. Anthem suit would fight final mega-merger hurdle. PMID- 15332522 TI - A commission out of joint. Another report adds to evidence that JCAHO needs a total overhaul. PMID- 15332523 TI - Pushing performance. When quality and safety aren't goals but preconditions for doing business. PMID- 15332524 TI - By popular demand. Quality focus, labor shortages drive pay, survey shows. PMID- 15332525 TI - He's the dean of CEOs. Harman has held the top job at his West Virginia hospital for nearly 40 years--and counting. PMID- 15332526 TI - The norm of self-interest. AB - The self-interest motive is singularly powerful according to many of the most influential theories of human behavior and the layperson alike. In the present article the author examines the role the assumption of self-interest plays in its own confirmation. It is proposed that a norm exists in Western cultures that specifies self-interest both is and ought to be a powerful determinant of behavior. This norm influences people's actions and opinions as well as the accounts they give for their actions and opinions. In particular, it leads people to act and speak as though they care more about their material self-interest than they do. Consequences of misinterpreting the "fact" of self-interest are discussed. PMID- 15332527 TI - The diversification of psychology: a multicultural revolution. AB - The National Multicultural Conference and Summit was held in January 1999 in Newport Beach, California. Hosted by Divisions 17 (Counseling Psychology), 35 (Society for the Psychology of Women), and 45 (Society for the Psychological Study of Ethnic Minority Issues), the event drew support from many American Psychological Association (APA) divisions and other major organizations and sponsors. Approximately 550 psychologists and graduate students attended the conference, which was intended to (a) examine state-of-the-art issues in ethnic minority psychology, (b) identify barriers to becoming a multicultural profession, and (c) forge alliances for political action and advocacy. The summit participants unanimously endorsed resolutions aimed at implementing cultural competence in all psychological endeavors. Multicultural themes arising from the summit included the diversification of the United States; the facilitation of difficult dialogues on race, gender, and sexual orientation; spirituality as a basic dimension of the human condition; the invisibility of monoculturalism and Whiteness; and the teaching of multiculturalism and diversity. APA was strongly encouraged to take the lead in seeing that multicultural competence becomes a defining feature of psychological practice, education and training, and research. PMID- 15332528 TI - Science, ethnicity, and bias: where have we gone wrong? AB - The quality, quantity, and funding of ethnic minority research have been inadequate. One factor that has contributed to this inadequacy is the practice of scientific psychology. Although principles of psychological science involve internal and external validity, in practice psychology emphasizes internal validity in research studies. Because many psychological principles and measures have not been cross-validated with different populations, those conducting ethnic minority research often have a more difficult time demonstrating rigorous internal validity. Thus, psychology's overemphasis of internal as opposed to external validity has differentially hindered the development of ethnic minority research. To develop stronger research knowledge on ethnic minority groups, it is important that (a) all research studies address external validity issues and explicitly specify the populations to which the findings are applicable; (b) different research approaches, including the use of qualitative and ethnographic methods, be appreciated; and (c) the psychological meaning of ethnicity or race be examined in ethnic comparisons. PMID- 15332529 TI - Test security: protecting the integrity of tests. PMID- 15332530 TI - Mistaken inferences. PMID- 15332532 TI - Toward more evidence-based guidelines for psychology. PMID- 15332533 TI - Why not test reading in three alphabets? PMID- 15332534 TI - [COX and study of cancer therapy]. AB - The increased expression of COX-2 in carcinoma tissue is found in gastric cancer, lung cancer and breast cancer as well as colon cancer. It was shown that COX-2 played an important role in carcinogenesis by an experiment using a cultured cell and an animal experiment using a knockout mouse. The inhibitory effect of COX-2 inhibitor on polyps in familial adenomatous polyposis (FAP) patients was reported from the clinical aspect, and the U. S. Food and Drug Administration (FDA) approved administration of a COX-2 inhibitor to FAP patients. COX-2 plays an important role in proliferation, invasion and metastasis of cancer. COX-2 expression was reportedly enhanced in lung cancer by an anticancer agent and radiotherapy, and clinical application of a COX-2 inhibitor is attempted. In addition, the experimental examination showed the inhibitory effect of a COX-2 inhibitor on hematogenous metastasis of colon cancer. The mechanism of the COX-2 inhibitor remains unclear. Clinical application of the COX-2 inhibitor to an effective anti-tumor agent is expected after more studies have been conducted on its molecular biologic function. PMID- 15332535 TI - [Case mix based payment and DPC from an international perspective]. AB - The basic principle of Diagnosis Related Groups (DRG) is to classify patients according to the combination of diagnosis and procedures. The first DRG developed by research at Yale University was adopted for the per-case-payment American Medicare scheme for hospital fees under a DRG/PPS (prospective payment system). This scheme has been refined continuously and adopted as payment systems in other countries. The DPC-based reimbursement scheme in Japan consists of two components; a DPC component and a Fee-For-Service (FFS) component. The DPC component corresponds to the "so-called" hospital fee, includes the hospital basic charge, the pharmaceuticals and supplies used in wards, lab-test, radiological examination, and any procedures costing less than 10,000 yen. The FFS component corresponds to the charges for surgical procedures, pharmaceuticals and supplies used in operation rooms, and procedures costing more than 10,000 yen. For the DPC component, a per diem payment schedule is set for each DPC group. One of the most important objectives in introducing the DPC scheme is to ameliorate the transparency of hospital activities in Japan. The DPC system is expected to make hospital services measurable, and then to provide a common ground for discussion among interest groups over the optimal policy to maintain a sustainable Japanese medical care system. However, in order to keep DPC more reliable, much must be done to refine the DPC version 3. For example, development of CCP (Co-morbidity-Complication-Procedure) matrix is now ongoing to more properly describe the resource consumption of each DPC group. It is also important how to evaluate nursing services, ICU services and expensive drugs and materials within the DPC sheme. PMID- 15332536 TI - [Management of hospitals in the prospective payment system]. AB - Since last year a prospective payment system, the so-called "diagnosis procedure combination" system has been implemented at 82 hospitals, and this fiscal year national universities and national hospitals became independent agencies. Furthermore, a new postgraduate training and education system started this year. Now it is time for hospitals to transform into institutions that are opted for by health professionals, patients, and medical students. Every hospital has to transform into a hospital that provides safe health care with a minimal number of medical errors and delivers care with a degree of information, transparency and logicality that will fully satisfy patients. That care must also be distinguished by efficiency giving proper consideration to costs. For this purpose, all hospital staff including physicians, nurses, technicians, pharmacists, dietitians, and clerical staff have to pursue health care as a team. In a comprehensive health care system, practice of team-based care is imperative. As we think that the implementation of critical paths (or clinical paths) will be a strong impetus for team-centered care and, especially important, for a change in the mindset of the physicians, we have addressed this subject. PMID- 15332537 TI - [Prospective payment system of medical cost and surgical cancer treatment]. AB - The process of introducing a prospective payment system after DRG or DPC in Japan was reviewed. The effect of DPC/PPS on surgical practice was discussed. Many problems resulted in the USA by introducing the DRG/PPS system. The rapid introduction of DPC/PPS in Japan only to reduce the total medical costs may bring about undesirable results for cancer patients. PMID- 15332538 TI - [DPC and its implications for clinical practice in cancer treatment]. AB - In April 2003, the Japanese government introduced a casemix classification based reimbursement methodology into acute inpatient care services by developing a new, national patient grouping, called Diagnosis Procedure Combination (DPC). The result, so far, has been successful, but analysis of data gathered using DPC revealed; 1) large deficit accumulating in short-term chemotherapy and examination admissions, 2) great variation in practice patterns within academic medical centers adopting the new system. Such phenomenon may lead to rapid shift of related services to outpatient setting, initiate debate over standardization of practice in cancer treatment. Efforts are needed to overcome these issues before the next reform planned in 2006. PMID- 15332539 TI - [Cancer chemotherapy in DPC]. AB - After the introduction of DPC in 2003, cancer chemotherapy treatment was moved to our outpatient clinic. We have initiated an outpatient cancer chemotherapy center, and the chemotherapy regimens were developed for various kinds of cancer: 1 for esophageal, 7 for gastric, 9 for colon, 38 breast and 10 for hepato-biliary pancreatic cancer. Since cancer chemotherapy on an outpatient basis is a world wide trend, we should maintain the quality of cancer chemotherapy for inpatients in our outpatient center. PMID- 15332540 TI - [The role of the critical path in a new hospital payment system according to a diagnosis procedure combination in Japan]. AB - Implementation of the critical path in the healthcare system facilitates standardization of medical practices, which improves both quality and safety management, shortens the length of hospital stays, and economizes on medical resources. In a new payment system introduced to university hospitals in Japan, hospital fees are charged by the day according to the diagnosis-procedure combination (DPC). To prepare for DPC, standardization of medical care, shortening the average length of stay within all hospitals, cost reduction, establishment of hospital networks, and an increase in new patients are critical issues. Cost management can be achived effectively by using the critical path. PMID- 15332541 TI - [Reliability at the National Cancer Institute-Common Toxicity Criteria version 2.0]. AB - We evaluated the reliability of CTC v 2.0 based on source documents and also studied the degree of inconsistency in toxicity grading. Five clinical research coordinators from the National Cancer Center Hospital independently reviewed source documents from 17 patients and graded toxicities in the following common adverse events: diarrhea, nausea, stomatitis/pharyngitis, vomiting, febrile neutropenia, infection, infection unknown source, and sensory neuropathy. If grading was already documented on the medical chart, it was masked so that the coordinator could perform the evaluation without information bias. After the completion of toxicity grading, the participating coordinators discussed each case, and a consensus was reached for final toxicity grading. The proportion of agreement for each toxicity criteria are as follows: diarrhea; 0.59 (95%CI 0.35 0.82), nausea; 0.47 (0.23-0.71), stomatitis/pharyngitis; 0.59 (0.35-0.82), vomiting; 0.71 (0.49-0.92), febrile neutropenia; 0.88 (0.73-1.04), infection; 0.82 (0.64-1.01), infection by unknown source; 0.82 (0.64-1.01), sensory neuropathy; 0.65 0.42-0.87). The cause of variability largely depended on the differences in individual clinical assessment, and misunderstanding of toxicity criteria by coordinators has been observed. Even in a single institution environment, variability exists in the toxicity assessment and grading. Good training and education on toxicity assessment using common criteria and development of translated manual, including the interpretation of criteria assessment, may help reduce variability. PMID- 15332542 TI - [Pilot-testing of the Japanese version of the EORTC QLQ-STO22 gastric cancer module]. AB - The pilot testing of the Japanese translation of the European Organization for Research and Treatment of Cancer (EORTC) gastric cancer module (QLQ-STO22) was done for 10 patients (6 men; 4 women) with gastric cancer. Age ranged from 23 to 73. Five patients had a performance status (PS) of 0, 4 had a PS of 2, and 1 had a PS of 4. Five had stage IV gastric cancer. It took 20 to 40 minutes for the patients to complete the questionnaire. All stated that it did not take long a time to complete, and that each question was easy to understand as a whole. They also mentioned that the questionnaire appropriately described and represented their health condition. However, it was revealed that some linguistic revisions should be made for 5 of the 22 questions to clarify their meanings by adding examples or replacing unclear words with clearer ones. The Japanese version of the QLQ-STO22 was finalized via those revisions. We are waiting for the result of the validation study in an international field (EORTC protocol number 15001/40003) with a larger number of gastric cancer patients using standardized psychometric testing. PMID- 15332543 TI - [Role of postoperative adjuvant chemotherapy with gemcitabine for pancreatic cancer: feasibility and anti-tumor effect]. AB - BACKGROUND: The feasibility and anti-tumor activity of gemcitabine in postoperative adjuvant chemotherapy were evaluated retrospectively. PATIENTS AND METHODS: Sixteen patients with advanced pancreatic cancer, who had a pancreatic resection with curative intent over the three years up to February 2003, were enrolled in this study. Aggressive surgery with dissection of para-aortic nodes and nerves around the superior mesenteric and celiac artery was carried out. After the operation, all patients have been given biweekly administration of 1,000 mg/m2 gemcitabine for more than 12 courses. RESULTS: The chemotherapy was well tolerated with only mild symptomatic and hematologic toxicities. Grade 3 adverse effects were observed in only 3 patients (19%); nausea and vomiting in 1 patient and leucocytopenia in 2 patients. The disease-specific cumulative survival rates were 81% at 1 year and 47% at 2 years, with a median survival of 20.4 months. The median disease-free interval was 16.8 months in all patients. CONCLUSIONS: Adjuvant systemic chemotherapy utilizing gemcitabine was feasible with acceptable adverse effects. Gemcitabine is a promising agent for the treatment of resectable advanced pancreatic cancer, and a randomized control trial is warranted for gemcitabine-based chemotherapy. PMID- 15332544 TI - [Combination chemotherapy with docetaxel+carboplatin in the treatment of cancer of the ovary and fallopian tube]. AB - Docetaxel and carboplatin were used as adjuvant chemotherapy or maintenance chemotherapy for cancer of the ovary and fallopian tube. Docetaxel 70 mg/m2 and carboplatin (area under the concentration-versus-time curve of 5) were administered intravenously every 3 weeks. Thirty-two patients (median age, 54 years) were assessable. We had objective responses from 5 of 7 (70%) assessable patients. Our toxicity findings included the following: grade 3 or 4 neutropenia (70% of courses); grade 3 or 4 leucocytopenia (35% of courses); hypersensitive reaction (25% of patients, none requiring discontinuation of therapy): edema (30% of patients): peripheral neuropathy (6% of patients). The combination of docetaxel and carboplatin is highly active in cancer of the ovary and follapian tube. The adverse effect was significant neutropenia, but peripheral neuropathy was rare. This regimen represents a reasonable first-line option for patients with ovarian cancer. PMID- 15332545 TI - [Recurrence of non-small cell lung cancer after successful treatment with gefitinib--report of three cases]. AB - Gefitinib, a first epidermal growth factor receptor (EGFR) inhibitor has been reported to be effective in chemotherapy-resistant non-small cell lung cancer. Gefitinib was also reported, however, to produce severe adverse effects, such as interstitial lung disease. Thus, clinical evaluation of gefitinib is still controversial and further studies are needed. We present 3 cases of non-small cell lung cancer (stage IV adeno-carcinoma) that showed recurrence after successful gefitinib treatment. Symptoms of all patients ameliorated in a week after the therapy, and radiograph and laboratory data improved. However, all showed recurrence in 3 to 7 months. After recurrence, patient 1 was re-treated with gefitinib at the request of the patient. Patient 2 has been on gefitinib. Patient 3 was treated with gefitinib, which was later replaced by another chemotherapy agent. After recurrence, however, no effective response was obtained in any of the 3 cases. There have been several reports of good response to gefitinib therapy at the first time in patients with advanced non-small cell lung cancer, but few reports of recurrence in patients after successful therapy with this agent. This report is thought to be important for the clinical evaluation of gefitinib and useful in terms of information about the resistance of gefitinib. PMID- 15332546 TI - [A case of recurrent pulmonary lymphoepithelioma-like carcinoma responding to treatment with CBDCA/paclitaxel combined chemotherapy]. AB - A 57-year-old female underwent left lower lung lobectomy and was histologically diagnosed as having lymphoepithelioma-like carcinoma. One year and 2 months after surgery, pleural thickening was recognized, so she was administered 4 courses of CBDCA/PTX combined chemotherapy. Toxicity associated with the chemotherapy was very mild. Pleural thickening and effusion disappeared after treatment, so this case was judged to be a complete response. She suffered from left chest pain before chemotherapy, which later lessened. She was thus able to stop taking NSAIDs. Because primary pulmonary lymphoepithelioma-like carcinoma is a rare tumor, there is no standard chemotherapy treatment. CBDCA/PTX combined chemotherapy is effective for pulmonary lymphoepithelioma-like carcinoma and shows good tolerability and improves QOL. PMID- 15332547 TI - [A case of gastric GIST treated preoperatively by imatinib mesylate]. AB - Gastrointestinal stromal tumor (GIST), the most common mesenchymal tumor of the human gastrointestinal tract, is thought to originate from the interstitial cells of Cajal. The mutation of c-kit, cording KIT, is essential in the development of GIST. Imatinib mesylate (IM), an agent for chronic myeloid leukemia, was reported to inhibit tyrosine kinase activity of KIT and to be highly effective for GIST. We report, here, a case of huge gastric GIST who underwent neoadjuvant therapy followed by surgical resection. The patient was a 62-year-old man with GIST in cardia (KIT+, CD34+, mitotic rate 5/50 HPF), whose chief complaint was general fatigue. Because the huge tumor, 7.5 cm in size, directly invaded the pancreas, total gastrectomy with distal pancreatosplenectomy was necessary for curative resection. IM was administered (400 mg/body/day) as a neoadjuvant treatment for down-staging of the tumor. Leucopenia (grade 2) and diarrhea (grade 1) were observed as the adverse effects of IM. Partial response was obtained. He underwent proximal gastrectomy without pancreatosplenectomy since CT no longer showed direct invasion to the pancreas. Histological examination of the resected specimen revealed the extensive degeneration of the tumor, in which tumor cells containing condensed nuclei had decreased remarkably. Interestingly, mitotic rate decreased to 0/50 HPF in the effective area of the resected specimen, indicating that recurrent risk might be decreased. A part of the viable tumor cells, however, had the same feature to that in the biopsied specimen before treatment. The results suggest that the heterogeneity of GIST induces different sensitivity to IM. The postoperative course was uneventful and no sign of recurrence was observed 3 months after surgery. Neoadjuvant therapy with IM may become a useful strategy for GIST, as it reduces the tumor size and decreases the recurrence rate. PMID- 15332548 TI - [TS-1 was prescribed for a patient with stomach cancer with peritoneal dissemination who survived for 3 years and 2 months]. AB - A 42-year-old female patient underwent total gastrectomy for gastric cancer (Borrmann's Type 3). Many rice-grain sized peritoneal metastases were observed in the transverse colon and mesenterium. The lesion was diagnosed as stage IV cancer and the degree of radical cure was determined to be C. Chemotherapy with TS-1 was administered postoperatively. In each cycle, the drug was administered at a daily dose of 100 mg for 4 weeks, followed by a drug-free period of 2 weeks. The adverse reactions were mild, and she underwent the 2nd and further courses of therapy on an outpatient basis. Since she had acute cholecystitis during the 12th course, the drug was withdrawn for 2 months. Thereafter, the drug was started again after resolution of the cholecystitis. At present, ie, 3 years and 2 months after the surgery, the patient is receiving the 23rd course of chemotherapy on an outpatient basis, and abdominal CT shows no evidence of increase in the peritoneal metastases, enlargement of the intraperitoneal lymph nodes, or ascites. PMID- 15332549 TI - [A case of paraaortic lymph nodes metastases responding to TS-1 following surgery for gastric cancer]. AB - A 58-year-old man underwent distal gastrectomy with a D2 lymph adenectomy for advanced gastric cancer (pStage IIIB). Seven months later, abdominal CT revealed multiple paraaortic lymph nodes metastases. Radiation therapy was not effective, so TS-1 chemotherapy was started (each treatment course consisted of daily oral administration of 100 mg TS-1 for 4 weeks followed by 2 drug-free weeks). The CT findings revealed that the metastatic lesion had shrunk markedly after the first course. A complete response was observed after the fifth course, and was maintained thereafter. The serum level of CEA decreased from 337 to 2.7 ng/ml after the third course, but gradually rose again and stayed between 30 and 50 ng/ml. Although the re-elevation of serum CEA level suggested the existence of a recurrent lesion, no sign of recurrence was found by radiographical or endoscopic examinations. Leukocytopenia and anemia (grade 2) were the only observed adverse effects. This patient continues to undergo outpatient treatment with good QOL. PMID- 15332550 TI - [A case of gastric cancer complicated with multiple lung metastases responding to TS-1]. AB - A 56-year-old gastric cancer patient with multiple lung metastases and dilated cardiomyopathy was treated by chemotherapy with TS-1. This case was judged to be unresectable. TS-1 (100 mg/body/day) was orally administered for 4 weeks followed by a drug-free 2-week period as 1 course. At the completion of 3 courses, the multiple lung metastases were assessed to show a complete response (CR). There were no side effects and no hospitalization. This chemotherapy is being continued, and a CR in the lung and no change of the gastric cancer have been maintained for 15 months (10 courses). PMID- 15332551 TI - [Weekly administration of paclitaxel might be beneficial against TS-1-resistant recurrent gastric cancer--a case report]. AB - At one year after postoperative adjuvant therapy consisting of TS-1, UFT, and PSK, lymph-node metastasis and recurrent suprarenal metastasis were confirmed in a patient with Stage IV gastric cancer. Despite TS-1 therapy (120 mg/body, 4 consecutive weeks, 2-week break), the therapeutic effect remained PR, and the severity of adverse reactions, such as skin symptoms, leukopenia, and diarrhea, ranged from grade 1 to 3. Although this therapy was continued intermittently, the therapeutic effect worsened to PD and, as a result, the therapy was discontinued. Next, in accordance with the treatment of recurrent and advanced breast cancer, weekly paclitaxel (PTX 80 mg/m2, 3 consecutive weeks, 1-week break) administration was performed on an outpatient basis. CR was achieved after 4 cycles. In addition, no adverse reactions were seen. Therefore, the weekly administration of PTX for the treatment of recurrent gastric cancer resistant to 5-FU anticancer agents is extremely useful, as the antitumor effect of this therapy was great, no adverse reactions were seen, and it can be performed on an outpatient basis. PMID- 15332553 TI - [A successfully resected case of colorectal cancer with multiple liver metastases treated with systemic chemotherapy]. AB - The patient was a-54-year-old man. Sigmoidectomy was performed for sigmoid colon cancer in 1991. Partial liver resection in 1992 and microwave coagulation therapy (MCT) in 1994 were carried out for liver metastasis. Complete remission of the metastasis was achieved. In June 2002, multiple liver and lung metastases were identified. 5-FU, CDDP therapy was applied as systemic chemotherapy. The liver metastasis was improved and the level of CEA was reduced. However, a new lesion appeared in the right lobe of the liver, followed by an increase of the CEA level. Although CPT-11, 5-FU, CDDP therapy was applied, it was not effective. The reason was considered to be from the decrease of drug delivery resulting from an unbalanced blood supply in the right lobe of the liver. Right lobectomy and radio frequency ablation was performed on June 17, 2003. The level of CEA was dramatically decreased. Additionally, CPT-11, 5-FU, CDDP therapy was applied. The patient has survived for 11 years after liver metastasis was first detected. PMID- 15332552 TI - [A case of hepatic metastasis from colon cancer successfully treated with 5-FU, levofolinate (l-LV) and low-dose CPT-11]. AB - The patient was a 67-year-old man in whom hepatic metastasis from transverse colon cancer was detected 15 months after transverse colectomy (D2). We treated the patient by systemically administering 2 courses of 5-FU 750 mg/day with l-LV 350 mg/day (once weekly for 6 weeks per course). Assessment of therapeutic effects by CT showed PD in the patient. As a second-line therapy, we treated the patient by systemically administering 3 courses of 5-FU 750 mg/day, l-LV 350 mg/day and CPT-11 40 mg/day x 3 days (once a week for 4 weeks per course). After 3 courses of this chemotherapy, CT examination revealed a reduction in the tumor size of the liver, and CEA levels decreased at the end of this chemotherapy. This chemotherapy also showed no high-grade toxicities. l-LV/5-FU/low-dose CPT-11 seems to be effective for metastatic colon cancer, and safe from the toxicity standpoint. PMID- 15332554 TI - [A case of liver metastasis of colon cancer responding remarkably to 5'-DFUR]. AB - A 65-year-old man was referred to our hospital because of diarrhea due to sigmoid colon cancer. Abdominal CT scan revealed a hepatic tumor (S8) about 2 cm in diameter. We performed a sigmoidectomy and planned to resect the liver metastasis 1 or 2 months later. Pathological findings showed moderately differentiated adenocarcinoma, s, n1. Two weeks after the surgery, 5'-DFUR was administered at 600 mg/day. An abdominal CT scan 2 months later demonstrated regression of the liver metastasis and another scan 4 months later showed the tumor had disappeared. 5'-DFUR was administered for about 2 years. Five years after the surgery, the patient is alive without recurrence and CEA level is in normal range. PMID- 15332555 TI - [Long-term control of rectal liver metastases by hepatic arterial infusion chemotherapy of 5-fluorouracil and l-leucovorin--a case report]. AB - For synchronous liver metastases from rectal cancer, after extirpation of the primary lesion, a transcatheter arterial embolization was postoperatively performed 3 times at 6-month intervals. Metastatic lesions in S1 and S8 resulted in progressive disease at 8 and 13 months postoperatively, and simultaneous sharp elevation of both CEA and CA19-9 levels. Therapy was then changed to hepatic arterial infusion (HAI) chemotherapy combined with 5-fluorouracil and l leucovorin (17 months postoperatively). S1 and S8 demonstrated CR and PR, respectively, 8 months after initiation of HAI. Correspondingly, both CEA and CA19-9 levels decreased dramatically, and have thereafter remained stable to date without marked elevation 36 months after the start of HAI. These findings suggest that our regimen for HAI chemotherapy may effectively control liver metastases and restrain progression over the long-term, that is, establish "tumor dormancy" as proposed in recent years. Moreover, CEA and CA19-9 played important roles as surrogate markers that reflect the response to metastatic tumors. PMID- 15332556 TI - [mRNA expression of chemokine receptors in hepatic and pancreatic tumor cell lines]. AB - Chemokines represent a large family of polypeptide signaling molecules that are notable for their role in chemotaxis, leukocyte homing, and directional migration. Recent observations have indicated that the expression of chemokine receptors on cancer cells may play a role in tumor progression and metastasis. In this study, the expression of mRNA for chemokine receptors in various human tumor cell lines was analyzed by multiplex-polymerase chain reaction (M-PCR). Strong expression of CCR6 mRNA in 3 of 3 hepatoma cell lines was observed. In the 3 pancreatic cancer cell lines, no specific expression of chemokine receptors was observed. Raji (lymphoma cell line) strongly expressed CCR7 and CXCR4. We further investigated CCR6 mRNA expression in these cell lines by real-time quantitative PCR. Similar results were obtained by both the PCR methods. Because human liver constitutively express liver and activation-regulated chemokine (specific ligand for CCR6), hepatoma cells may selectively root and spread in the liver. Strong CCR7 and CXCR4 expressions in the lymphoma cell may explain the organ specificity of lymphoma for lymphoid organs as well. These findings probably indicate that some cancer cells have organ specificity via expression of chemokine receptors. PMID- 15332557 TI - [Key issues in sentinel node biopsy for breast cancer]. AB - Whereas the majority of surgeons in Western countries perform sentinel node biopsy (SNB) for early breast cancer, the majority of Japanese surgeons do not. Veronesi very recently reported the results of a clinical trial in which SNB without axillary lymph node dissection for small breast cancer did not increase axillary recurrence. Thus, sentinel node surgery has been accepted as a safe and accurate method of screening the axillary nodes for metastasis in women with small breast cancer. SNB should thus be recommended as standard care for early breast cancer in Japan. SNB for patients with ductal carcinoma in situ (DCIS) may be performed according to the decision of doctors or patients in each case, because the indication of SNB for DCIS is controversial. Since preoperative chemotherapy could increase the rate of false-negative sentinel nodes because of the induced lymphatic changes, SNB is thought to be safer before than after preoperative chemotherapy. Current evidence does not allow internal mammary SNB to be recommended as a standard procedure, but as patients with internal mammary node involvement may benefit from adjuvant systemic treatment, internal mammary SNB should be further studied in this context. Preoperative diagnosis of an axillary metastasis using fine-needle aspiration cytology (FNAC) under ultrasonographical imaging or core needle biopsy under MR imaging can cost effectively decrease the indications of SNB. PMID- 15332558 TI - [Gastrointestinal cancer]. AB - Although their sensitivity is not high, SCC, TPA and IAP are useful for esophageal cancer. The sensitivity of CEA, CA 19-9, is relatively high, especially in well-differentiated adenocarcinoma of gastric cancer with lymph node metastasis. AFP is specific to liver metastasis from gastric cancer, and CA 125 is also specific to peritoneal dissemination. CA 72-4 and NCC-ST-439 are useful markers for advanced staging. CEA, CA 19-9, is useful for colon cancer, especially for predicting preoperative staging. Half-life and doubling time of tumor markers is useful in some cases for the evaluation of operation and chemotherapy. We showed our data concerning postoperative CEA and/or CA 19-9 monitoring after operation for gastric cancer in 120 recurrent patients. Positivities of CEA and CA 19-9 for recurrence were 65.8% and 85.0%, respectively, both of which were significantly higher than the preoperative sensitivities (28.3% and 45.0%, respectively). In most patients with high levels of preoperative CEA and/or CA 19-9, these tumor markers increased again at recurrence. Recurrent diseases were detected between 5 months after detection by diagnostic imagings and 12 months before detection by diagnostic imagings (mean of 3.1+/-3.6 months before detection by diagnostic imagings) and between 10 months after detection by diagnostic imagings and 13 months before detection by diagnostic imagings (mean 2.2+/-3.9 months before detection by diagnostic imagings) by CEA and CA 19-9 monitorings, respectively. These results suggest that CEA and/or CA 19-9 monitoring after operation was useful to predict the recurrence of gastric cancer, especially in almost all the patients with high preoperative levels of these markers. PMID- 15332560 TI - When no treatment is good treatment. PMID- 15332559 TI - The 3rd Conference on Asian Trends in Prostate Cancer Hormone Therapy. AB - From its inception in 2001, the Conference on Asian Trends in Prostate Cancer Hormone Therapy has served as an annual forum for Asian urologists to compare data on prostate cancer and to discuss issues regarding the use of hormone therapy. The 3rd conference, held in Tokyo in December 2003, began with participants from China, Indonesia, Japan, Korea, Singapore, and Taiwan presenting QOL data deriving from a survey of patients with prostate cancer. For this purpose, each country translated the Functional Assessment of Cancer Therapy Prostate (FACT-P) questionnaire into its own language. Although the surveys conducted in each country included a heterogeneous cohort of patients and gave very mixed results, the trial of FACT-P in Asian countries seemed propitious and in future may provide insights that could prove beneficial to patients. Day 2 of the conference included 2 discussions, focusing on the most appropriate number of biopsy cores and the implementation of prospective trials involving the collaboration of Asian countries, respectively. In the latter discussion, although a varied assortment of proposals were put forth, the participants generally agreed that any collaborative study must be a prospective outcome study conducted in a relatively short time not exceeding 2-3 years, and that patient registration should be done using the Internet. PMID- 15332561 TI - CDHM observance comes up with winning idea. PMID- 15332562 TI - Emergency plan development. Guiding principals underlying policies with respect to planning for catastrophic events and public health threats. PMID- 15332563 TI - Inflammation, cardiovascular disease and destructive periodontal diseases. The evolving role of the dental profession. AB - Destructive periodontal diseases have been associated with increased risk of atherosclerotic complications, including myocardial infarction (MI) and stroke. This finding comes at a time when our understanding of atherosclerotic complications are changing from a focus on the occlusion of arteries, due to the buildup of plaque deposits, to an increased awareness of the role of inflammation in plaque rupture and thrombus formation. The role of inflammation can have great significance to the dental profession if inflammatory cells and factors derived from chronic infections, such as destructive periodontal diseases, are shown to contribute to plaque rupture. This "Perspectives" feature will review the role of inflammation in atherosclerotic complications, and the association between destructive periodontal diseases, systemic inflammation and atherosclerotic complications. It will also highlight ongoing research designed to determine whether destructive periodontal diseases contribute to atherosclerotic complications. PMID- 15332564 TI - A survey of floss frequency, habit and technique in a hospital dental clinic & private periodontal practice. AB - The purpose of the study was to determine floss frequency, habits and techniques of patients entering a private periodontal office (office site) vs. a hospital dental clinic (hospital site). It was found that the flossing frequency in a hospital dental clinic is far less than in a private practice, and patients who do floss may not be using the proper flossing technique. Patients need more instruction and care with regard to their interproximal oral hygiene. PMID- 15332565 TI - Fiber-reinforced composite bridge and splint. Replacing congenitally missing teeth. AB - The congenital absence of permanent teeth, also known as partial anodontia, is a relatively common problem. The patient missing one or both lateral incisors is a frequent occurrence. The treatment of missing laterals involves both functional and esthetic considerations. Options include: cuspid lateralization; implant placement; conventional three-unit bridge; Maryland bridge; and a fiber reinforced composite bridge (direct or indirect). Direct fiber-reinforced composite bridges provide a minimally invasive restorative option that allows for future implant placement. Selection of appropriate fiber reinforcement and placement of the fibers allows long-term clinical success. Adherence to appropriate adhesive protocol is mandatory. PMID- 15332566 TI - The supplementary role of panoramic radiographs in diagnosis and prevention of life-threatening systemic health condition. Case report. AB - A 71-year-old white male had a routine panoramic radiograph taken for oral health reasons. Positive carotid calcifications findings on the radiograph were in agreement with significant carotid stenosis as determined by duplex ultrasonic examination. If the occasion arises, the authors advocate that a routine panoramic radiograph examination include an assessment of the region of the carotid system for presence of carotid calcifications in all adult patients. PMID- 15332567 TI - Destination: Tanzania. NYU professor leads dental mission to Africa to offer free care to destitute population. PMID- 15332568 TI - Self-funded plans. Finding the right medical and dental policy for employees involves matching the ability to pay to the proper benefit package. PMID- 15332569 TI - [Outlook from the past to the future]. PMID- 15332570 TI - [First results of stenting of bifurcation stenoses of the coronary arteries]. AB - AIM: To study effectiveness of various types of coronary stenting in bifurcation stenoses. MATERIAL AND METHODS: Fifty six patients with bifurcation stenoses were divided into 2 groups: stenting of the basic and side branch of the coronary artery (CA) was made in group 1 patients (n = 27), stenting of the basic branch and balloon angioplasty of the side branch were made in group 2 (n = 29). Primary clinical and angiographic response reached 96 and 90% in groups 1 and 2, respectively. Complications were absent in group 1, occurred in 13.7% cases in group 2 (the difference was insignificant). One year after the procedure lethal outcomes were absent. The number of unfavourable clinical outcomes was higher in group 1 (25.9 and 13.7%, respectively, p = 0.4). The rate of restenoses detected angiographically was not significantly different in group 1 and 2 (33 and 26%, respectively). CONCLUSION: Stenting of bifurcation stenoses brings about good short-term and satisfactory long-term results in most of the patients. The rate of the complications was low. Two stents implantation (in basic and side branches) has no significant advantages over stenting of the basic CA and balloon angioplasty of its side branch. PMID- 15332571 TI - [Prevention of cardiovascular diseases in Russia: successes, failures, prospects]. PMID- 15332572 TI - [New approaches to the treatment of ischemic heart disease: therapeutic angiogenesis in combination with surgical revascularization of the myocardium]. AB - AIM: To evaluate the results of surgical treatment of patients with ischemic heart disease in combination with intraoperative intramyocardial introduction of the human gene VEGF165 (angiostimulin). MATERIAL AND METHODS: Twenty four patients enrolled in the study were examined using 12-lead ECG, echocardiography, treadmill exercise test, single-photon emission computed tomography of the myocardium with Tc-99m-tetrophosmine, fluorodesoxyglucose (FDG) positron-emission tomography of the myocardium, selective coronarography. Out of 24 patients, 10 patients have been reexamined so far. The effect of the treatment was assessed by the data obtained at the treadmill test, transthoracic echocardiography, myocardial scintigraphy and FDG. RESULTS: All the patients demonstrated a clinical response. The class of effort angina improved, the dose of the prescribed nitric drugs was decreased, exercise tolerance and total stress time rose, quality of life improved. Myocardial scintigraphy registered reduction of the total area and better accumulation of the radiopharmaceutical under load and at rest 3 and 6 months after the operation including myocardial areas which had not been revacularised at coronary artery bypass grafting but had been treated with the preparation of the human gene VEGF165. CONCLUSION: The results of the study suggest that therapeutic angiogenesis may be an alternative impact on those myocardial areas which are supplied by the affected but ineligible for bypass grafting coronary arteries. PMID- 15332574 TI - [Ethnic differences in apolipoprotein B levels in hyperlipidemia in Russian and Kirghiz populations]. AB - AIM: To evaluate the levels of apolipoprotein B (apo-B) and coronary heart disease (CHD) risk factors in Russians and the Kirghiz with primary hyperlipidemia (PHL). MATERIAL AND METHODS: Lipid spectrum of the blood (LDLP and HDLP cholesterol, total cholesterol, triglycerides), apo-B, CHD risk factors were studied in 60 Russian and 75 Kirghis patients with PHL aged 28 to 67 years (mean age 50.0 +/- 7.59 years, 56 females and 79 males). RESULTS: In Russians a mean level of LDLP cholesterol and apo-B was significantly higher than in Kirghiz patients (3.95 +/- 1.29 mmol/l vs 3.51 +/- 1.17 mmol/l, p = 0.042; 173.3 +/- 57.3 mg/dl vs 145.5 +/- 49.3 mg/dl, p = 0.003, respectively). CHD in both ethnic groups occurred with similar rates. The multifactorial regression analysis shows that an apo-B concentration can serve an independent risk factor associated with CHD in the Kirghiz population (beta = 0.25, p = 0.03) while in Russians living in Kirghizia this factor is a low content of HDLP cholesterol (beta = -0.25, p = 0.05). PMID- 15332573 TI - [Polymorphic markers of GNB3 (C825T), AGTR1 (A1166C) and ACE (A2350G and I/D) genes in patients with arterial hypertension combined with diabetes mellitus type 2]. AB - AIM: To elicit correlations of polymorphic markers of GNB3 (C825T), AGTR1 (A1166C), ACE (A2350G and I/D) genes with arterial pressure, left ventricular hypertrophy (LVH) and blood concentrations of proinflammatory cytokines in hypertensive patients with diabetes mellitus type 2 (DM2). MATERIAL AND METHODS: Clinical parameters (24-h arterial pressure profile, echocardiographic findings, immunoenzymes level) were studied in 89 hypertensive patients with DM2. These patients had different genotypes by the studied allele variants of the genes determined by polymerase chain reaction. RESULTS: Polymorphism of A1166C gene of type 1 vascular receptor of angiotensin II (AGTR1) contributes to formation of arterial hypertension (AH) signs diversity in DM2 patients. GNB3, a gene C825T polymorphic marker, showed a correlation with diastolic arterial pressure but this variant of the gene locus is not associated with LVH. However, G-allele of ACE gene contributes much to appearance of this pathological sign. Mean values of IL-1beta and TNF-alpha as well as the presence of LVH depended on genotypes by ACE gene (polymorphism I/D). CONCLUSION: Polymorphic markers of ACE and GNB3 candidate genes influence clinical diversity of pathological signs in DM2 patients through modification of AH and LVH severity and the level of proinflammatory cytokines. PMID- 15332575 TI - [Cardiorenal interactions: clinical implication and role in pathogenesis of cardiovascular and renal diseases]. PMID- 15332576 TI - [Detection of sodium monourate crystals in biopsies of gastric mucosa in patients with gout]. AB - AIM: To examine gastric biopsies with polarization microscopy for detection of sodium monourate crystals (SMC). MATERIAL AND METHODS: The trial included 20 patients with gout diagnosis (mean age 55.7 years, mean duration of the disease 12.3 years) in whom esophagogastroduodenoscopy was made with biopsy of gastric mucosa from the antral part of the stomach and middle third of the gastric body. RESULTS: Crystals in the biopsy specimens were detected in 11 of 20 examinees. The crystals were characterized by strong double refraction, length 3-20 mcm, acicular or planiform shape, blue or yellow color depending on position in compensated polarized light. Quantitative distribution of the crystals within one biopsy specimen was uneven and varied from solitary crystals to clusters of 70-80 crystals in sight, up to formation of tophus-like structures. Clinical picture in detection of SMC was characterized by more frequent occurrence of cases with subcutaneous tophuses of various location combined with higher hyperuricemia. CONCLUSION: One of the essential lines in the research of gout concerns mechanisms of SMC formation in organs and tissues and microcrystalline gastroduodenal inflammation. Methods of correction of this inflammation are to be designed. PMID- 15332577 TI - [Molecular, genetic, and cellular aspects of the heart and vessel remodeling in hypertension (review)]. PMID- 15332578 TI - [First experience of clopidogrel application in the treatment of acute myocardial infarction with ST-elevation]. AB - AIM: To examine efficacy of clopidogrel before thrombolytic therapy (TLT) in patients with acute myocardial infarction (AMI). MATERIAL AND METHODS: A total of 72 patients (48 males and 24 females) admitted to hospital within 6 hours since the onset of acute myocardial infarction (AMI) were divided into three groups. Group 1 and 2 patients (n = 38 and 20, respectively) were given a prehospital stress dose of aspirin (250-500 mg), group 3 patients (n = 14) received this dose at admission. TLT with streptokinase or actilise (1,500,000 IU and 100 mg, respectively) was given to all the patients. Before TLT group 2 received a stress dose of clopidogrel (300 mg), after TLT--75 mg/day. 12-Lead ECG, CFK activity and troponine content examinations were made. The patients were observed for 30 days. The recovery of coronary circulation (CC) was evaluated by dynamics of a total lowering of the elevated ST segment. RESULTS: CC completely recovered 60 min after TLT in 16, 10 and 7% patients of groups 1, 2 and 3, respectively. CC recovered partially in 24, 55 and 7% patients, respectively. In 90 min after TLT partial recovery of CC was observed in 21, 31 and 43%, respectively; in 180 min the effect was absent in 19, 15 and 38%, respectively. 30-Day lethality was 11.1% and was high in group 3. No lethal outcomes were seen in group 2. CONCLUSION: It is possible to achieve better myocardial reperfusion after TLT and improve 30-day outcomes in MI patients given combined antithrombocytic therapy. PMID- 15332579 TI - [Correlation of lipemia level after fat loading with manifestation of atherosclerosis in coronary arteries]. AB - AIM: To examine relationships between lipemia, atherogenicity of blood lipoproteins spectrum after fat loading (FL) and severity of angiographic manifestations of coronary atherosclerosis. MATERIAL AND METHODS: The study enrolled 72 males free of ischemic heart disease (IHD)--group 1; 60 IHD patients with moderate affection of the coronary arteries (CA), i.e. maximal narrowing of one CA 50%--group 2; 107 IHD patients with severe CA affection, i.e. CA narrowing > 50%--group 3. FL was given by J. R. Patsch technique, blood sampling was made before meal, 3 and 6 hours after FL. RESULTS: It was shown that intake of food fats in IHD and IHD-free patients stimulated development of postprandial lipemia of different severity and duration. It appeared due to increased fraction of lipoproteins rich in triglycerides. In group 2 and 3 there was a definite and up to 6 hours in duration postprandial hypertriglyceridemia associated with proatherogenic deviations in lipid and protein components (LDL and HDL) by FL hour 6: growing of LDL cholesterol, apoB, apoB/apoAI and lowering of HDL cholesterol and apoAI. Patients with severe CA atherosclerosis (> 50% narrowing of three CA) had the most severe atherogenic affections in the postprandial lipoprotein spectrum. CONCLUSION: Postprandial lipemia/hypertriglyceridemia and atherogenic changes in the spectrum of lipoproteins after FL correlate directly with angiographic manifestations of coronary atherosclerosis. PMID- 15332580 TI - [Hyperhomocysteinemia and acute phase proteins in various forms of ischemic heart disease]. AB - AIM: To determine clinical significance of high concentrations of homocystein, C reactive protein, fibrinogen in various forms of ischemic heart disease. MATERIAL AND METHODS: Enzyme immunoassay was made to measure serum concentrations of homocystein, C-reactive protein, fibrinogen in 60 patients with ischemic heart disease (IHD) in the form of stable effort angina (n = 20), painless myocardial ischemia (n = 19), unstable angina pectoris (n = 21) and 20 control patients free of IHD. Myocardial ischemia was confirmed at dobutamine stress echocardiography. RESULTS: Serum concentrations of homocysteine, C-reactive protein and fibrinogen were higher in patients with unstable angina than in the other examinees with IHD. A statistically significant correlation exists between homocysteine serum levels and acute phase proteins (C-reactive protein, fibrinogen) in patients with unstable angina. In the other groups it was absent. CONCLUSION: Correlation between serum levels of homocysteine and acute phase proteins in patients with unstable angina suggests a direct participation of this amino acid in destabilization of atherosclerotic plaques and development of acute coronary syndromes. PMID- 15332581 TI - [Characteristics of atherosclerotic lesion of the abdominal aorta and its unpaired visceral branches in patients with chronic abdominal ischemia]. AB - AIM: To study characteristics of echocardiographically detectable structural changes of the abdominal major arteries due to atherosclerosis with consideration of clinical symptoms. MATERIAL AND METHODS: 174 participants of the study had chronic abdominal ischemia (CAI). 20 healthy individuals served as control. The analysis was made of the data obtained at clinical laboratory tests, endoscopic and x-ray examinations, ultrasound investigation (USI) of the abdominal organs, USI of abdominal arteries in B-mode, transabdominal USI of the vertebral column. RESULTS: Echographically detectable signs of atherosclerotic affection of abdominal arteries in CAI patients are the following: increased lumen of the abdominal aorta (AA), unpaired visceral arteries in proximal and/or distal parts, relative narrowing of unpaired visceral arteries at the site of their origin from the aorta, lengthening of the abdominal aorta, S-shape configuration of the aorta on the longitudinal sections, thick aortal wall, unpaired visceral arteries, uneven thickness of the wall, plane foci of high echogenicity on the internal surface of the vascular wall, single or multiple hyperechogenic foci responsible for partial obstruction in the vessels. CONCLUSION: Degenerative changes of the intervertebral disks and corresponding atherosclerotic changes of the abdominal aorta, atherosclerotic affection of the unpaired visceral arteries may cause abdominal pain syndrome in CAI patients. PMID- 15332582 TI - [Cerebrovascular complications in metabolic syndrome: possible approaches to decrease risk]. AB - AIM: To compare brain perfusion in hypertensive patients with diabetes mellitus type 2 (DM2) or metabolic (MS) syndrome and hypertensive patients without clinicobiochemical signs of DM2 or MS; to study enoxaparin effects on brain perfusion in DM2 and arterial hypertension (AH). MATERIAL AND METHODS: Seventy patients included in the study were divided into three groups: 30 patients with DM2 and AH (group 1), 30 patients with MS and AH (group 2) and 10 AH patients without manifestations of MS or DM2 (group 3). All the patients have undergone single-photon emission computed tomography (SPECT) of the brain, carbohydrate and lipid metabolism were examined. RESULTS: Deterioration of brain perfusion was more prominent in DM2 and MS patients with AH than in hypertensive patients with normal metabolism. Stress test with acetasolamide revealed defective autoregulation of cerebral blood flow in hypertensive patients with DM2. A 6-week therapy with enoxaparin significantly improved brain perfusion in hypertensive patients with DM2. CONCLUSION: Enoxaparin treatment of hypertensive DM2 and MS patients with abnormal perfusion of the brain can be used for prevention of cerebrovascular complications. PMID- 15332583 TI - [Characteristics of hypotensive effect in patients with arterial hypertension and desaturation signs of obstructive sleep apnea syndrome during sleep]. AB - AIM: To estimate the efficacy of 8-week antihypertensive monotherapy in patients with arterial hypertension (AH) regarding the presence of obstructive sleep apnea syndrome (OSAS). MATERIAL AND METHODS: We analysed the results of 24-h blood pressure (BP) monitoring of 26 inpatients (mean age 54 +/- 2 years) with mild (n = 18) and moderate (n = 8) AH before and after 8 weeks of treatment with 5-10 mg amlodipine or 50-100 mg of losartan once daily to assess blood pressure profile parameters. The patients underwent nocturnal monitoring of arterial oxygen saturation (pulsoximeter NONIN-8500 M, USA). The presence of OSAS was confirmed when a characteristic clinical picture was combined with the presence of significant (> 4%) sleep desaturation episodes > 15 episodes per hour or the presence of group desaturation episodes below 90%. Seven hypertensive patients with OSAS were assigned to group 1, nineteen patients without OSAS--to group 2. The differences in estimated parameters between the groups were tested by Mann Whitney U test, the dynamics of BP profile parameters--by Wilcoxon matched pairs test. RESULTS: In group 1 there were no significant differences by most of BP profile parameters before and after antihypertensive treatment, except mean nocturnal systolic BP. In group 2 a significant hypotensive effect was seen by all parameters of BP profile except BP variability. Hypotensive efficacy in group 2 was 1.5-2 times higher vs group 1, but the difficulties were not significant. CONCLUSION: Antihypertensive therapy in hypertensive patients with OSAS is less effective than in those without OSAS but it is not uneffective. PMID- 15332584 TI - [Blockers of angiotensin receptors: a novel approach to the treatment of secondary pulmonary hypertension]. AB - AIM: To evaluate efficacy of losartan, a blocker of angiotensin receptors, in combined treatment of secondary pulmonary hypertension (SPH) in patients with chronic obstructive bronchitis (COB). MATERIAL AND METHODS: Losartan effects on hemodynamics, blood gases and clinical course of the disease were studied in 29 patients with COB and SPH (mean age 52 +/- 1.7 years). A control group consisted of 15 patients (mean age 51 +/- 1.5 years) treated with cardiac glycosides and diuretic drugs. M- and B-mode Doppler echocardiography registered hemodynamic parameters. 24-h monitoring of AP and ECG were made by standard methods. Blood gases and venous rheology were examined. RESULTS: Losartan administration in COB patients with SPH improved hemodynamics. Stroke index rose from 36.3 +/- 2.1 to 45.8 +/- 2.1 ml/m2 (by 26.2%, p < 0.01) in SPH functional class III, from 26.3 +/ 1.9 to 32.7 +/- 2.1 ml/m2 (by 24.3%, p < 0.01) in functional class IV Cardiac index rose by 22.2 and 21.1%, respectively. Pulmonary hemodynamics improved too: systolic pressure in pulmonary artery fell by 25.7% in functional class III, by 18.6% in functional class IV. Losartan normalized a 24-h AP profile, reduced the number of painless myocardial ischemia. CONCLUSION: Use of losartan in combined therapy of patients with COB and SPH improves clinical status of the patients, corrects basic cardiohemodynamic parameters, has a positive effect on AP profiles without negative impact on blood gas composition and rheology. PMID- 15332585 TI - [Amiodarone (cordarone) efficiency in atrial extrasystole]. AB - AIM: To assess antiarrhythmic amiodarone efficacy of suppressing atrial extrasystoles resistant to other antiarrhythmic modalities. MATERIAL AND METHODS: The antiarrhythmic effect of amiodarone was studied in 70 patients (38 males, 32 females, mean age 49.6 +/- 1.7 years, mean duration of arrhythmia 4.9 +/- 1.5 years). The loading oral dose of amiodarone was 600-1200 mg/day for 10 days, mean maintenance dose--1656.25 mg a week, mean duration of treatment--27.5 +/- 3.2 months. The response to amiodarone was estimated by repeated 24-h Holter monitoring. RESULTS: A complete response was observed in 78.5% patients in loading and in 65.7% patients in maintenance therapy. A partial antiarrhythmic response was seen in 8.57 and 16.41% patients, respectively. CONCLUSION: Amiodarone is a basic drug against ventricular arrhythmia. Its effectiveness in suppression of atrial extrasystole is weaker but also appropriate. PMID- 15332586 TI - [Cardiac arrest in a patient with obstructive sleep apnea during sleep]. PMID- 15332587 TI - [Pacemaker syndrome in the absence of artificial pacemaker (pseudopacemaker syndrome)]. PMID- 15332588 TI - [Acquired deficiency in the system of proteins C and S and acute myocardial infarction]. PMID- 15332589 TI - [Endothelium--a new target for a therapeutic action of angiotensin-converting enzyme inhibitors]. PMID- 15332591 TI - Staphylococcus aureus adhesion to different treated titanium surfaces. AB - Staphylococcus aureus is a major pathogen, associated with medical-device related infections. Converting biomaterial surfaces into non-interactive surfaces requires a specific surface/interface design. One approach is to polish the surface, and a second is to coat the surface with an antimicrobial or protein resistant coating. This study showed that polishing a titanium surface or coating titanium with various treatments that decreased the surface's coefficient of friction, had no significant effect on minimising S. aureus adhesion to these surfaces under static conditions in comparison to standard medical grade titanium. The cell promoting coating, TAST, was found to increase the S. aureus density on its surface as expected. The only coating that significantly decreased the density of adhering S. aureus was the titanium surface coated with sodium hyaluronate. Thus such a coating could have potential use as a coating for ostoesynthesis, orthopaedic or dental implants. PMID- 15332590 TI - Innovative tissue engineering structures through advanced manufacturing technologies. AB - Awide range of rapid prototyping (RP) techniques for the construction of three dimensional (3-D) scaffolds for tissue engineering has been recently developed. In this study, we report and compare two methods for the fabrication of poly (epsilon-caprolactone) and poly-(epsilon-caprolactone)-poly-(oxyethylene)-poly (epsilon-caprolactone) copolymer scaffolds. The first technique is based on the use of a microsyringe and a computer-controlled three-axis micropositioner, which regulates motor speed and position. Polymer solutions are extruded through the needle of the microsyringe by the application of a constant pressure of 10-300 mm Hg, resulting in controlled polymer deposition of 5-600 microm lateral dimensions. The second method utilises the heating energy of a laser beam to sinter polymer microparticles according to computer-guided geometries. Materials may be fed either as dry powder or slurry of microparticles. Both powder granulometry and laser working parameters influence resolution (generally 300 microm x 700 microm), accuracy of sintering and surface and bulk properties of the final structures. The two RP methods allow the fabrication of 3-D scaffolds with a controlled architecture, providing a powerful means to study cell response to an environment similar to that found PMID- 15332592 TI - The expression of CD44 in archival paraffin embedded interface tissues of failed orthopaedic implants. AB - CD44 hyaluronan receptor is present on large number of different cell type. It acts as one of the adhesion proteins, binding to hyaluronan and is known to play a part in cell migration from vessels in inflammation. The aim of this study was to examine the presence and distribution of CD44 in interface membrane in aseptic loosening. Immunohistochemistry (IHC) using human anti-mouse CD44 antibody studied 20 aseptically loosened interface samples. Extracted protein from all cases was examined by Western blot and RT-PCR. CD44 was detected in 85% of interfaces by IHC and the presence of protein confirmed by blotting and RT-PCR, which showed the mRNA level for CD44. CD44 was expressed by macrophage, multinucleated giant cells, mast cells and lymphocytes. Further studies are needed to characterise the role of this molecule in the inflammatory response to wear debris in aseptic loosening. PMID- 15332593 TI - Effects of nano-scaled particles on endothelial cell function in vitro: studies on viability, proliferation and inflammation. AB - Recent studies give support for a connection between the presence of inorganic particles (of microm and nm size) in different organs and tissues and the development of inflammatory foci, called granulomas. As the potential source of particles (e.g. porcelain dental bridges) and the location of particle detection were topographically far apart, a distribution via the blood stream appears highly probable. Thus, endothelial cells, which line the inner surface of blood vessels, would come into direct contact with these particles, making particle endothelial interactions potentially pathogenically relevant. The objective of this study was to evaluate the effects that five different nano-scaled particles (PVC, TiO2, SiO2, Co, Ni) have on endothelial cell function and viability. Therefore, human endothelial cells were exposed to different amounts of the above mentioned particles. Although most particle types are shown to be internalised (except Ni-particles), only Co-particles possessed cytotoxic effects. Furthermore, an impairment of the proliferative activity and a pro-inflammatory stimulation of endothelial cells were induced by exposure to Co- and, to a lesser extent, by SiO2-particles. If a pro-inflammatory stimulation of endothelial cells occurs in vivo, a chronic inflammation could be a possible consequence. PMID- 15332594 TI - PIRAC Ti nitride coated Ti-6AI-4V head against UHMWPE acetabular cup-hip wear simulator study. AB - Wear behaviour of TiN(titanium nitride)-coated Ti and Ti-6AI-4V alloy against UHMW polyethylene was studied in hip simulation test. Ti alloys possess an excellent combination of mechanical properties and biocompatibility, however, they suffer from inadequate wear resistance. Thus, their use as articulating components of total joint replacements requires surface hardening, e.g. by TiN. Thirty-two millimetre diameter cp-Ti and Ti-6AI-4V femoral heads were coated with several micrometre thick TiN layers employing an original PIRAC nitriding method based on interaction of Ti alloy substrate with highly reactive monatomic nitrogen. The heads were tested against UHMWPE cups at 37 degrees C in Ringer's solution or in distilled water. Simulator tests were performed at peak pressures of 1.5 and 2.0 MPa in a constant rotation mode at the frequency of 1.5 Hz. The wear of UHMWPE was estimated by weight loss, and the worn metallic and polyethylene surfaces were examined in SEM. The wear rate of UHMWPE cups articulating against PIRAC coated Ti and Ti-6AI-4V after up to 4 x 10(6) cycles was significantly lower than that of UHMWPE articulating against 316L stainless steel. No delamination of TiN coatings was observed after 4 x 10(6) cycles. These results suggest that TiN PIRAC coating on Ti-6AI-4V heads could minimise the wear of total hip replacements without compromising the mechanical properties of the femoral component. PMID- 15332595 TI - Culture of haematopoietic cells in a 3-D bioreactor made of Al2O3 or apatite foam. AB - Foams of Al2O3 and apatite ceramics with interconnecting pores were produced using a new technique. The surfaces of the ceramics served as substrates for the culture of human peripheral and bone marrow derived stem cells. Up to 27 days the cells were kept in culture where they proliferated and developed into different morphologies consistent with bone marrow cell lines. PMID- 15332596 TI - Immobilisation of catalase on the surface of biodegradable starch-based polymers as a way to change its surface characteristics. AB - In this study, a specific enzyme catalase was immobilised onto the surface of two different biodegradable materials, starch cellulose acetate (SCA) and starch polycrapolactone (SPCL) blends. This immobilisation was achieved by several different routes, mainly by covalent binding and an adsorption method using as activation agents epichlorohydrin, cyanogen bromide (CNBr), and aminopropyltriethoxysilane. The effect of the coupling pH of the enzyme-support reaction was determined in terms of activity recovery (%). The catalase immobilised on SCA showed higher activity recovery (%) for all the methods used as compared with results obtained with SPCL. The immobilisation process using epichlorohydrin as an activation agent and polyethylenimine as a spacer-arm enhanced the stability and the half-lives at pH 7.0, 30 degrees C, for immobilised catalase on both SCA and SPCL. The half-lives were respectively, 1162 and 870 h compared with other treatments and free enzyme (480 h). The free glycerol present in the immobilisation medium was also a factor that contributed toward the better performance regarding the long-term stability at 30 degrees C and neutral pH. The extension of the morphological modifications on the surface of the materials was observed by scanning electron microscopy. In general, the results indicated that the chemical modification with epichlorohydrin could provide a simple and rather efficient technique to modify the starch-based materials' surface that might be useful in several biomedical applications. PMID- 15332597 TI - Water as foaming agent for open cell polyurethane structures. AB - The problem of moisture in polymer processing is known to any polymer engineer, as air bubbles may be formed. Hence granulates are generally dried prior to manufacturing. This study tried to develop a novel processing methods for scaffolds with controlled moisture content in thermoplastic polyurethane. The common foaming agents for polyurethane are organic solvents, whose residues remaining in the scaffold may be harmful to adherent cells, protein growth factors or nearby tissues. Water was used as a foaming agent and NaCl was used as porogens to achieve an open-cell structure. The polyether-polyurethane samples were processed in a heated press, and achieved a porosity of 64%. The pore size ranged between 50 and 500 microm. Human fibroblasts adhered and proliferate in the scaffold. A non-toxic production process was developed to manufacture a porous structure with a thermoplastic polyether-polyurethane. The process enables a mass-production of samples with adjustable pore size and porosity. In contrast to an existing method (solvent casting), the processing of the samples was not limited by its thickness. The process parameters, which attribute mostly to the pore building, were filling volume, temperature, NaCl-concentration and water uptake rate. PMID- 15332598 TI - Controlling the supramolecular organisation of adsorbed collagen layers. AB - The supramolecular organisation of collagen adsorbed on polymer substrates was investigated as a function of properties of the substrates (chemical nature, roughness) and of characteristics of the collagen solution (concentration, state of aggregation) as well as details of the preparation procedure (adsorption time, drying rate). Elongated structures are formed at the interface by assembly of collagen molecular segments protruding into the solution. This is favoured by using a hydrophobic and smooth substrate, by increasing the adsorbed amount and by increasing the adsorption time, even beyond stages at which the adsorbed amount does no longer vary. Collagen adsorbed at low amount on hydrophobic substrates strongly reorganises into a net-like pattern if drying is performed at low rate. This is due to dewetting and collagen displacement by the water meniscus. Applications derived from the control of collagen organisation are presented. Nanostructured polymer surfaces were created starting from a collagen template. The attachment and the cytoskeletal organisation of mammalian cells (MCF-7/6) were also shown to depend on collagen organisation. PMID- 15332599 TI - The effect of silica nanoparticulate coatings on cellular response. AB - The current techniques used to create patterned materials at the nanometer scale such as electron beam lithography are restricted to patterning small areas, which can be expensive and time consuming. A simple, cost-effective approach has been developed to create a reproducible surface topography to influence the cellular response. In this study, the cellular response of murine fibroblasts to 7, 14 and 21 nm colloidal silica particles were investigated over one, three and seven days and up to seven weeks. The surface topography and wettability of the surfaces were also studied. The results confirmed that silica particles create a nanoscale topography, which initiates a distinctive cellular response affecting the morphology, adhesion and proliferation of the fibroblasts. The effect was evident up to seven weeks with no adverse effects on cell viability. PMID- 15332600 TI - Relationship between bioceramics sintering and micro-particles-induced cellular damages. AB - We performed experimental studies to confirm the hypothesis that cellular damages occurring around implanted biphasic bioceramics could be related to a micro particles release because of an insufficient sintering. First, an in vitro cytotoxicity study was performed on four biphasic ceramic (BCP) samples. Without treatment of the extraction medium, a cytotoxicity was observed, although after centrifugation this cytotoxicity disappeared in all samples. Second, micro particles of hydroxyapatite (HA), beta-tricalcium phosphate (beta-TCP) and 40% beta-TCP/60%HA mixture were used for a cell inhibition study. A decrease of cell viability was observed with the increase in particles concentration. At 10000 particles per cell, the viability and proliferation were completely inhibited. Third, HA, beta-TCP and BCP ceramic granules were implanted in rabbit femoral cavities for 12 weeks. No degradation of HA granules was observed. The degradation was higher for beta-TCP (40%) than for BCP (5%). On the other hand, new bone formation was significantly higher for beta-TCP (21%) and HA (18%) than for BCP (12%). More micro-particles were formed around BCP granules than around beta-TCP, and phagocytised by macrophages. The release of ceramic micro-particles could be related to the sintering process. BCP ceramic have to be sintered at only 1160 degrees C. Consequently, HA micro-particles of BCP ceramic are incompletely sintered and easily released after immersion or implantation. The microparticles could be at the origin of local inflammation and cell damage and could perhaps modify osteogenesis. Attention must be paid to this problem especially with BCP ceramics because of the sintering difficulties of this bioceramic. PMID- 15332601 TI - Immunohistological identification of receptor activator of NF-kappaB ligand (RANKL) in human, ovine and bovine bone tissues. AB - Receptor activator of NF-kappaB ligand (RANKL, also called ODF/TRANCE/OPGL) is the final factor of osteoclast differentiation. Osteoclastogenesis may be determined by its receptor RANK and the relative ratio of RANKL to its decoy receptor osteoprotegerin (OPG), and alterations in this ratio may be a major cause of bone loss in many metabolic and immunologic disorders. In order to get a better insight into this complex regulatory system, this study aimed to determine where RANKL protein is located in bone tissues. RANKL was stained immunohistochemically in ex vivo human, ovine and bovine bone tissue. RANKL was observed labelled in the membrane of osteoblasts, osteocytes and osteoclasts and their surrounding matrix. In cartilage, which was used as a negative tissue control, chondrocytes were not stained. The presence of RANKL protein in the membrane of osteoblasts and also the secretion of RANKL by osteoclasts has been hypothesised in earlier studies. In this study, RANKL protein was shown histologically for the first time in the membrane and in the long processes of osteocytes. The result strongly suggests the crucial involvement of osteocytes in terms of orchestrating bone remodelling by influencing differentiation and activation of osteoclasts. PMID- 15332602 TI - Influence of octacalcium phosphate coating on osteoinductive properties of biomaterials. AB - In this study, we investigated the influence of octacalcium phosphate (OCP) coating on osteoinductive behaviour of the biomaterials. Porous titanium alloy (Ti6Al4V), hydroxyapatite (HA), biphasic calcium phosphate (BCP) and polyethylene glyco terephtalate/polybuthylene terephtalate (PEGT-PBT) copolymer, all uncoated and coated with biomimetically produced OCP, were implanted in back muscles of 10 goats for 6 and 12 weeks. Uncoated Ti6Al4Vand HA did not show any bone formation after intramuscular implantation. All OCP coated implants, except PEGT-PBT, did induce bone in the soft tissue. The reason for the non-inductive behaviour of the copolymer is probably its softness, that makes it impossible to maintain its porous shape after implantation. Both uncoated and OCP coated BCP induced bone. However, the amount of animals in which the bone was induced was higher in the coated BCP implants in comparison to the uncoated ones. Osteoinductive potential of biomaterials is influenced by various material characteristics, such as chemical composition, crystallinity, macro- and microstructure. OCP coating has a positive effect on osteoinductivity of the biomaterials. The combination of the advantages of biomimetic coating method above traditional methods, and a good osteoinductivity of OCP coating that is produced by using this method, opens new possibilities for designing more advanced orthopaedic implants. PMID- 15332603 TI - Creep-resistant porous structures based on stereo-complex forming triblock copolymers of 1,3-trimethylene carbonate and lactides. AB - Stereo-complexes (poly(ST-TMC-ST)) of enantiomeric triblock copolymers based on 1,3-trimethylene carbonate (TMC) and L- or D-lactide (poly(LLA-TMC-LLA) and poly(DLA-TMC-DLA)) were prepared. Films of poly(ST-TMC-ST) could be prepared by solvent casting mixtures of equal amounts of poly(LLA-TMC-LLA) and poly(DLA-TMC DLA) solutions and by compression moulding co-precipitates. Although compression moulding was performed at 191 degrees C, thermal degradation was not apparent and materials with good tensile properties could be obtained. For compression-moulded poly(ST-TMC-ST) specimens containing approximately 16 mol % lactide, the values for E-modulus, yield stress and elongation at break were respectively 17, 1.7 MPa and 90%. Also a very low long-term creep rate of 2.2 x 10(-7)s(-1) was determined when specimens were loaded to 20% of the yield stress. When compared with compression-moulded poly(TMC), poly(ST-TMC-ST) specimens deform at a rate that is one to two orders of magnitude lower. Furthermore, poly(ST-TMC-ST) specimens showed complete dimensional recovery within 24 h after loading to 20% and 40% of the yield stress for 40 and 5.5 h, respectively. Highly porous poly(TMC) and poly(ST-TMC-ST) structures with interconnected pores were prepared by a method combining co-precipitation, compression moulding and salt leaching. After prolonged compressive deformation, solid and porous poly(ST-TMC-ST) discs showed significantly better recovery behaviour than poly(TMC) discs. PMID- 15332604 TI - Control of fibronectin displacement on polymer substrates to influence endothelial cell behaviour. AB - Thin layered biomaterial surfaces of maleic anhydride copolymers are provided as a versatile platform for biomaterial applications. The provided comonomers define the character of the surface and its behaviour towards biomolecules and biosystems, such as proteins and cells. The kinetics of adsorption, desorption, and exchange of fibronectin and human serum albumin were investigated on different copolymer surfaces. Two different species of adsorbed proteins were found, a fast and a slow desorbing one. Furthermore, the exchange process depends on the kind of pre-adsorbed protein and the kind of exchange protein, as well as of the hydrophobicity of the copolymer surface. In this context adhesion, proliferation, and differentiation of endothelial cells from the umbilical cord vein onto fibronectin pre-coated surfaces were studied. Strong correlation between fibronectin exchange characteristics and the formation of focal adhesions, reorganisation of fibronectin, and generation of vascular-like structures by the cells was observed. PMID- 15332605 TI - Metabolic and histological analysis of mesenchymal stem cells grown in 3-D hyaluronan-based scaffolds. AB - Sheep mesenchymal stem cells (MSCs) were isolated and expanded using the principle of plastic adherence. Their identity as progenitor cells was confirmed by induction along the osteoblastic lineage using osteogenic supplements and observation of calcific deposits by von Kossa staining. MSCs were seeded onto two types of hyaluronan-based cylindrical scaffolds in high concentrations and cultured for varying time points up to three weeks. Culture medium was supplied using the following conditions: statically, on a shaker, by stirring with a magnetic stirrer or by perfusion in a tubular flow circuit. Total cell metabolism was assessed by MTT assay and the quality of cell coverage and matrix formation observed by SEM and histological analysis of thin sections of the constructs. Perfusion culture was established as the most appropriate culturing conditions, with cell metabolism increasing by approximately 300% over three weeks. The coverage of the scaffold surface was very good and the deposition of collagenous matrix was superior in these conditions compared to the, static and other dynamic culture conditions. PMID- 15332606 TI - Heterogeneity in proliferative potential of ovine mesenchymal stem cell colonies. AB - Bone marrow biopsies were taken from the iliac crest of 28 individual sheep from three different breeds, ranging in age from 4 months to 8 years and mesenchymal stem cells (MSCs) isolated using selection due to plastic adherence. Cells were cultured in medium that had been selected for its effect on observed MSC proliferation, until populations of greater than 50 million had been obtained from each biopsy. The identity of the isolated cell populations as progenitors of the mesenchymal lineage was verified by deriving both osteoblastic and chondrocytic phenotypes when cultured in osteogenic and chondrogenic medium supplements, respectively. The rate of cell proliferation for each marrow biopsy was measured at each passage and the number of initial stem cells in each sample estimated. There was no statistically significant correlation between the age of the sheep and MSC proliferative potential, or age and estimated initial MSC number. There was no apparent significant difference between proliferation rate and sheep breed and colonies established from frozen cells grew at similar rates to pre-frozen cells. Counter intuitively, there appeared to be a negatively correlated trend between proliferation rate and MSC concentration in the samples. It is concluded that no initial descriptive statistics of the marrow biopsies can assist in estimating the proliferative potential, and therefore the timing of future surgeries, of MSCs sampled for the purposes of tissue engineering. PMID- 15332607 TI - Bacterial biosynthesis of a calcium phosphate bone-substitute material. AB - A species of Serratia bacteria produces nano-crystalline hydroxyapatite (HA) crystals by use of a cell-bound phosphatase enzyme, located both periplasmically and within extracellular polymeric materials. The enzyme functions in resting cells by cleaving glycerol-2-phosphate (G-2-P) to liberate free phosphate ions which combine with calcium in solution to produce a cell-bound calcium phosphate material. Bacteria grown as a biofilm on polyurethane reticulated foam cubes were challenged with calcium and G-2-P in a bioreactor to produce a 3-D porous bone substitute material. The scaffold has 1 mm macropores and 1 microm micropores. XRD showed the crystallites to be 25-28 nm in size, resembling HA before sintering and beta-tricalcium phosphate (beta-TCP, whitlockite) after. When biofilm was grown on titanium discs and challenged with calcium and G-2-P, a calcium phosphate layer formed on the discs. Biomineralisation is therefore a potential route to production of precursor nanophase HA, which has the potential to improve strength. The scaffold material produced by this method could be used as a bone-filler or as an alternative method for coating implants with a layer of HA. PMID- 15332608 TI - Cements from nanocrystalline hydroxyapatite. AB - Calcium phosphate cements are used as bone substitute materials because they may be moulded to fill a void or defect in bone and are osteoconductive. Although apatite cements are stronger than brushite cements, they are potentially less resorbable in vivo. Brushite cements are three-component systems whereby phosphate ions and water react with a soluble calcium phosphate to form brushite (CaHPO4 x 2H2O). Previously reported brushite cement formulations set following the mixture of a calcium phosphate, such as beta-tricalcium phosphate (beta-TCP), with an acidic component such as H3PO4 or monocalcium phosphate monohydrate (MCPM). Due to its low solubility, hydroxyapatite (HA) is yet to be reported as a reactive component in calcium phosphate cement systems. Here we report a new cement system setting to form a matrix consisting predominantly of brushite following the mixture of phosphoric acid with nanocrystalline HA. As a result of the relative ease with which ionic substitutions may be made in apatite this route may offer a novel way to control cement composition or setting characteristics. Since kinetic solubility is dependent on particle size and precipitation temperature is known to affect precipitated HA crystal size, the phase composition and mechanical properties of cements made from HA precipitated at temperatures between 4 and 60 degrees C were investigated. PMID- 15332609 TI - The effect of surface treatment of hydroxyapatite on the properties of a bioactive bone cement. AB - Bioactive bone cements based on a paste-paste system for orthopaedic applications have been developed. They consist of hydroxyapatite (HA) filler particles in a methacrylate matrix comprising urethane dimethacrylate (UDMA) and triethylene glycol dimethacrylate (TEGDMA). To improve the interface between inorganic filler and organic matrix the HA particles were subjected to two different surface treatment methods, using polyacrylic acid (PAA) and gamma methacryloxypropyltrimethoxy silane (gammaMPS). The aim of the present study was to determine the influence of surface treatment on the mechanical properties, namely compressive strength (CS), diametral tensile strength (DTS) and three point flexural strength (FS) of the cements and the effect of ageing in simulated body fluid (SBF). Comparing the mechanical properties of the two cements after fabrication, the gammaMPS-HA cement showed higher strength values for all tests conducted (CS = 185+/-19.6 MPa, DTS = 27+/-2.5 MPa, FS=50.2+/-4.9 MPa), whereas PAA-HA containing cement had strength values around 20% lower. However, poly(acrylic acid) surface treatment was found to be more effective in improving the interface, and PAA-HA cements maintained their mechanical properties after immersion in SBF whereas gammaMPS-HA cement showed a reduction in strength values post ageing. From the results of this study, it is concluded that PAA treatment of the HA filler is a viable alternative to silanation with gammaMPS which may provide increased durability in aqueous environments. PMID- 15332610 TI - Development and cell response of a new biodegradable composite scaffold for guided bone regeneration. AB - Composites of biodegradable polymers with different calcium phosphate ceramics and glasses, have been developed as scaffolds for applications in bone-tissue engineering. In this work, phosphate glass particles have been incorporated into the polymer, poly(95L/5DL) lactic acid (PLA) and porous structures were elaborated. Their porosity, compressive mechanical properties and biological response were evaluated. Interconnected structures with evenly distributed pores and a porosity as high as 97% were obtained. The incorporation of glass particles into the polymer showed to have a positive effect in the mechanical properties of the foams. Indeed, the compressive modulus increased from 74.5 to 120 KPa and the compressive strength from 17.5 to 20.1 KPa for the PLA and the PLA/glass foams, respectively. The biological response was evaluated by means of the MTT test, the materials resulted to be noncytotoxic. PMID- 15332611 TI - Design, synthesis and properties of a degradable polyurethane scaffold for meniscus regeneration. AB - Longitudinal lesions in menisci are among the most frequent orthopedic problems of the knee. Repair by simple techniques is only limited to the vascular part of the meniscus. For repair of the avascular part of the meniscus a scaffold, which will assist the body in the formation of new meniscus cell tissue, might be applicable. In this study a biomedical segmented polyurethane with poly(epsilon caprolactone) as soft segment and 1,4-butanediisocyanate and 1,4-butanediol as uniform hard segments has been synthesised. The material has a micro phase separated morphology and excellent mechanical properties. A porous scaffold was prepared via a combination of liquid-liquid phase separation and salt leaching. The foams prepared combined a very high interconnectivity and porosity with the desired compression modulus. After six months of implantation in the knees of beagles full ingrowth with cells was obtained and it was found that meniscus like tissue had been formed in the scaffold. Moreover, compression behaviour appeared to be comparable to native meniscus tissue. PMID- 15332612 TI - Investigating cell-material interactions by monitoring and analysing cell migration. AB - Cell-material interactions can on one hand be characterised by assessing the functional state and or shape of the cells at one or different discrete periods of time, on the other hand by observing cell migration and spreading behaviour. The object of this study was to investigate the migration behaviour of fluorescently labelled cells, and to evaluate the software analysing this migration. In the present study, the behaviour of fibroblasts cells on differently structured surfaces was taken as example. In the first step, the influence of seven different lipophilic dyes (Dil, DiO, DiA, DiD, DiR, PKH2 and PKH26) on cell performance was determined taking biochemical parameters as indices. In the second step, the fluorescence characteristics of these dyes were compared regarding their applicability. In the third step, migration behaviour of Dil-labelled fibroblastic cells on plane and grooved surfaces were monitored and analysed using specific software. Our data suggest that most of the dyes have optimal characteristics for studying cell-cell interactions. Cell migration behaviour regarding migration direction and cell spreading was different on plane and grooved surfaces. It could be shown that computer-based image analysis represents a practical, quick and objective tool to quantify exactly cell migration behaviour. PMID- 15332613 TI - Cellular investigations on electrochemically deposited calcium phosphate composites. AB - Electrochemically deposited calcium phosphate (CaP) coatings are fast resorbable and existent only during the first period of osseointegration. In the present study, composite coatings with varying solubility (hydroxyapatite (HA), brushite with less HA and monetite (M) with less HA) were prepared and the influence of the degradation and the reprecipitation of CaP on osteoblastic cells were investigated. On the brushite composite coating a new precipitated, finely structured CaP phase was observed during immersion in cell culture medium with or without osteoblastic cells. The surface morphology of monetite and HA coatings were entirely unmodified under the same conditions. So it could be assumed that electrochemically deposited brushite with less HA acts as a precursor for new precipitated CaP. On this surface osteoblastic cells revealed a well-spread morphology with pronounced actin cytoskeleton and demonstrated good proliferation behaviour. Thus we suggest that brushite seems to be especially suitable for coating of implants as a matrix for nucleation and growth of new bone. PMID- 15332614 TI - In vitro assessment of the biological response to nano-sized hydroxyapatite. AB - Nano-sized, rod-like hydroxyapatite (nHA) crystals were produced and shown to be phasepure by X-ray diffraction analysis, as no secondary phases were observed. The nHA suspension was electrosprayed onto glass substrates using a novel processing routine to maintain nanocrystals of hydroxyapatite. The biocompatibility of nHAwas determined using human monocyte-derived macrophages and human osteoblast-like (HOB) cell models. The release of lactate dehydrogenase (LDH) from human monocyte-derived macrophages was measured as an indicator of cytotoxicity. The release of the inflammatory cytokine, tumour necrosis factor alpha (TNF-alpha) from cells in the presence of nHA crystallites was used as a measure of the inflammatory response. Although there was some evidence of LDH release from human monocyte-derived macrophages when in contact with high concentrations of nHA crystals, there was no significant release of TNF-alpha. Moreover, nHA-sprayed substrates were able to support the attachment and the growth of HOB cells. These results indicate that nHA crystals may be suitable for intraosseous implantation and offers the potential to formulate enhanced composites for biomedical applications. PMID- 15332615 TI - Cell interactions with laser-modified polymer surfaces. AB - The performance of a polymeric biomaterial depends on the bulk and surface properties. Often, however, the suitability of the surface properties is compromised in favour of the bulk properties. Altering the surface properties of these materials will have a profound effect on how cells and proteins interact with them. Here, we have used an excimer laser to modify the surface wettability of nylon 12. The surface treatment is rapid, cost-effective and can cause reproducible changes in the surface structure of the polymers. Polymers were treated with short wavelength ( < 200 nm) UV light. These wavelengths have sufficient photon energy (6.4eV) to cause bond scission at the material surface. This results in a surface reorganisation with incorporation of oxygen. Surface wettability changes were confirmed using contact angle measurements. Cell interactions with the surfaces were examined using 3T3 fibroblast and HUVEC cells. Cells morphology was examined using a confocal laser scanning microscope (CLSM). Cell activity and cell number on the treated nylon were assessed using biochemical assays for up to seven days. Both fibroblasts and endothelial cells initially proliferated better on treated compared with untreated samples. However, over seven days activity decreased for both cell types on the control samples and endothelial cell activity and cell number also decreased on the treated polymer. PMID- 15332616 TI - Phosphoserine--a convenient compound for modification of calcium phosphate bone cement collagen composites. AB - Temporary bone replacement materials on the basis of calcium phosphates and hydroxyapatite (HAP) are used in surgery for filling bone defects. Components which are able to control the nucleation and crystal growth of HAP through their functional groups and which can additionally activate bone cells may be helpful in the development of materials with enhanced remodelling in vivo. In this study, the influence of O-phospho-L-serine (PS) on the materials properties of calcium phosphate bone cement composites was investigated. For up to an addition of 25 mg/g PS a strong increase in the stability of the cements under load was determined. The material was studied by scanning electron microscopy and transmission electron microscopy. A more dense microstructure and a plate-like morphology of the HAP-crystals were detected in the modified composites compared with the non-modified samples. By X-ray powder diffraction an inhibition of the dissolution of alpha-tricalcium phosphate (alpha-TCP) and dicalciumphosphate anhydrous (DCPA) particles was found. alpha-TCP and DCPA are the main constituents of the cement precursor. The results of cell culture studies using rat calvaria osteoblasts demonstrate a good viability of the cells on the PS modified material. Furthermore, the proliferation and differentiation were found to be enhanced on the PS-modified material. PMID- 15332617 TI - Biocompatible Nb2O5 thin films prepared by means of the sol-gel process. AB - Thin biocompatible oxide films with an optimised composition and structure on the surface of titanium and its alloys can improve the implant integration. The preparation of these thin oxide layers with the intended improvement of the surface properties can be realised by means of the sol-gel process. Nb2O5 is a promising coating material for this application because of its extremely high corrosion resistance and thermodynamic stability. In this study, thin Nb2O5 layers ( < 200 nm) were prepared by spin coating of polished discs of cp-titanium with a sol consisting of a mixture of niobium ethoxide, butanol and acetylacetone. The thickness, phase composition, corrosion resistance and the wettability of the oxide layers were determined after an optimisation of the processing parameters for deposition of oxide without any organic impurities. The purity of the oxide layer is an important aspect in order to avoid a negative response to the cell adhesion. The biocompatibility of the oxide layers which was investigated by in vitro tests (morphology, proliferation rate, WST-1, cell spreading) is improved as compared to uncoated and TiO2 sol-gel coated cp titanium concerning the spreading of cells, collagen I synthesis and wettability. PMID- 15332618 TI - Preparation and physico-chemical characterisation of microporous polysaccharidic hydrogels. AB - A new technique to obtain microporous hydrogels was realised. It permits us to obtain a microporous structure directly on the already cross-linked hydrogel. It consists in stratifying the already cross-linked hydrogel on to a filter with known porosity and forcing the CO2 bubbles, derived from the addition of HCl to a porogen salt (NaHCO3), to cross through the filter first and then the matrix. By changing the porosity of the filter, it was possible to modulate the porous morphology of the hydrogels. The polysaccharides selected were hyaluronane, alginate, and carboxymethylcellulose. The influence of the porous morphology on the physico-chemical properties of the gel has been evaluated by FT-IR, FRAP, calorimetric, water uptake, and rheological analysis. PMID- 15332619 TI - Detection of micro- and nano-sized biocompatible particles in the blood. AB - The research deals with new scanning electron microscopic evaluations of the interface between blood and explanted temporary vena cava filters from patients affected by blood disorders. The biological tissues adherent to the filter and the small thrombi formed in vivo were detached from the metallic structure of the device, fixed, dehydrated and prepared for the histological and the electron microscopy. The analyses showed that both samples (thrombus and newly formed tissue) contained foreign, in some cases nano-sized, bodies. The chemistry of these particles was different and varied, and unusual compounds containing non biocompatible elements like bismuth, lead, wolfram, tungsten were also detected. The interaction between these debris travelling in the blood stream and the blood itself leads to suspect that the formation of the thrombus can originate from these inorganic and inert foreign bodies that act as triggering agent of the blood coagulation. PMID- 15332620 TI - In vitro host response assessment of biomaterials for cardiovascular stent manufacture. AB - The deployment of a vascular stent during angioplasty has greatly reduced the risks of restenosis. However, the presence of the device still induces a host response as well as a mechanical action on the blood vessel wall and an alteration of the haemodynamics. Platelet and inflammatory cells can adhere on the stent surface and be activated to produce biochemical signals able to stimulate an excessive proliferation of the smooth muscle cells with the consequent obstruction of the vessel lumen. For these reasons, the host response to two of the materials used in stent manufacture, stainless steel and diamond like carbon, was investigated in vitro. The data showed that stainless steel induced a higher level of host response both in terms of platelet aggregation and macrophage activation. However, the spreading of inflammatory cells was more accentuated on diamond-like carbon. The inflammatory cells produced levels of platelet-derived growth factor, a key signal in smooth muscle cell proliferation, similar to stainless steel thus suggesting that carbon coatings may not be able to prevent restenosis. PMID- 15332621 TI - Design and bioproduction of a recombinant multi(bio)functional elastin-like protein polymer containing cell adhesion sequences for tissue engineering purposes. AB - Genetic engineering techniques were used to design and biosynthesise an extracellular matrix (ECM) analogue. This was designed with a well-defined molecular architecture comprising different functional domains. The structural base is a elastin-derived repeating unit, which confers an adequate elastic characteristic. Some of these elastin domains have been modified to contain lysine; this amino acid can be used for crosslinking purposes. The polymer also contain periodically spaced fibronectin CS5 domains enclosing the well-known cell attachment sequence REDV. Finally, the polymer has target sequences for proteolitic action. These sequences are those found in the natural elastin and are introduced to help in the bioabsorption of the polymer. In addition, these proteolitic sequences were chosen in a way that, after proteolitic action, the released fragments will be bioactive. These fragments are expected to promote cell proliferation activity, angiogenesis and other bioactivities of interest for tissue growing, repairing and healing. After purification, the resulting polymers proved to be of high purity and correct sequence. Glutaraldehyde has shown to be a cross-linking agent for this polymer, yielding insoluble hydrogel matrices. This work is framed in a long term project aimed to exploit the power of genetic engineering for the design and bioproduction of complex ECM analogues showing the rich complexity and multi (bio)functionality of the natural matrix. PMID- 15332622 TI - Mechanical and leakage behaviour of the dentin--adhesive interface. AB - Dentine bonding systems (DBS) have been developed in order to bond restorative materials (i.e. composite) to the inner walls of the tissues when function and integrity as to be restored. Adhesion to dentine results from the penetration of DBS into the demineralised substrate constituted by a swollen collagen network. The short-term stability of a restored tooth is mainly affected by the presence of defects which act as stress raiser, while the long-term stability of a restored tooth is mainly affected by the seal of the restorative material on the dental structures. In order to determine the properties of the material interface, bonding to dentine is analysed using micro-tensile static and dynamic tests, assisted by the finite element modelling (FEM) and by the X-ray computed microtomography (micro-CT). The effect of voids and porosity in the composite layer of the DBS on the stress distribution has been investigated. Tensile adhesive strength for a particular DBS was measured on cylindrical specimens. The dual energy absorption technique, with the synchrotron beam light, has been developed to investigate, in a non-destructive manner, the leakage at the dentine DBS interface of a silver nitrate staining solution as a function of mechanical cycling. The results indicate that leakage occurs radially through the dentine adhesive interface and is influenced by the porosity in the adhesive and composite layers. PMID- 15332623 TI - Surface modified superparamagnetic nanoparticles for drug delivery: interaction studies with human fibroblasts in culture. AB - The concept of drug delivery using magnetic nanoparticles greatly benefit from the fact that nanotechnology has developed to a stage that it makes possible not only to produce magnetic nanoparticles in a very narrow size distribution range with superparamagnetic properties but also to engineer particle surfaces to provide site specific delivery of drugs. The size and surface characteristics of the nanoparticles are crucial factors that determine the success of the particles when used in vivo. The aim of this study was to modify the surfaces of the magnetic nanoparticles with PEG to improve the biocompatibility of the nanoparticles by resisting protein adsorption and increasing their intracellular uptake. In this study, the poly(ethyleneglycol) (PEG) modified superparamagnetic iron oxide nanoparticles have been prepared and their influence on human dermal fibroblasts is assessed in terms of cell adhesion/viability, morphology, particle uptake and cytoskeletal organisation studies. Various techniques have been used to determine nanoparticle-cell interactions including light, fluorescence, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The modification of nanoparticle surface induced alterations in cell behaviour distinct from the unmodified particles, suggesting that cell response can be directed via specifically engineered particle surfaces. PMID- 15332624 TI - Effects of serum and serum heat-inactivation on human bone derived osteoblast progenitor cells. AB - Generally, heat inactivated foetal calf serum (FCS) containing media are used for the cultivation of animal and human cells. The role of serum source and serum treatment on the behaviour of cells has long been neglected. The present study was performed to investigate the effects of serum heat inactivation and serum source on trabecular bone derived progenitor cells (HBC). Furthermore, it was investigated in how far these reactions differed from those seen in bone marrow derived mesenchymal progenitor cells (HBMC) cultures. We found that HBC cultures performed differently in the presence of FCS and HS with or without heat inactivation. The reactions similar to some degree those observed in HBMC cultures. The implications of the results on cell-implant surface interaction studies are discussed. PMID- 15332625 TI - Hydrophilic matrices to be used as bioactive and degradable bone cements. AB - Two different hydrophilic systems were investigated regarding their suitability to be used as enzymatically degradable and highly bioactive bone cements. They contained either acrylic acid (AA) or 2-hydroxyethyl methacrylate (HEMA) as the hydrophilic monomer. Swelling, degradation, mechanical and bioactivity tests were employed to characterise their behaviour. Although both of the systems were very hydrophilic, only the one containing HEMA was able to form an apatite-like layer on its surface. Moreover, this system could be degraded by amylolytic enzymes at a rate easily controlled by the incorporation of different amounts of enzyme to the formulation, as shown by the evolution of the mechanical properties, weight loss and glucose concentration in the solution. These results show these novel systems have a great potential to induce bone ingrowth inside the pores created during the degradation of the material, therefore establishing a strong interface with the tissue. PMID- 15332626 TI - Hydrophilicity of 3-D biomaterials: the Washburn equation. AB - Characterisation and quantification of the surface energy of biomaterials used as tissue engineering scaffolds is important, but many of the techniques available to examine these properties are only applicable to smooth flat samples, not porous materials. This paper describes the application of the Washburn equation to measure the surface energy of a range of porous polyether polyurethane scaffolds with three test liquids; n-Hexane was used to measure a material constant, whilst ethanol and xylene were used to measure contact angles. The results show that the Washburn equation is not applicable in its current form, reasons for this could be that the voids in the materials are too wide for effective capillarity; absorption of the solvents into the polymer matrix may further complicate the measured imbibition profile. Another possible reason is the differences between the sizes of the interconnecting pores in scaffolds with varying void sizes; this could affect the capillary effect of the test liquids through the material. The repeatability of the results and the similar patterns observed with the different liquids suggest that if these issues could be quantified and incorporated into the Washburn equation, it may be possible to generate useful results for similar materials. PMID- 15332627 TI - Rationalising the design of polymeric thermoresponsive biomaterials. AB - We investigated the cell adhesion and growth of a series of thermoresponsive copolymers of N-isopropylacrylamide (NIPA) and N-tert-butylacrylamide (NtBA) above their lower critical solubility temperatures (LCST). It was found that cell adhesion and growth on the solvent cast films improved with increasing the NtBA content in the copolymers. The improvement was dependent on cell line. The surfaces of copolymers were analysed by atomic force microscopy. The topography of polymer films was not dependent on composition. The differences in the cell attachment and growth were attributed to the variation of surface energy with composition. The surface energy of copolymers decreased with the increase in the NtBA content. We conclude that poly(N-isopropylacrylamide) (poly(NIPA)) is a relatively poor substrate for cell growth and proliferation. However, its ability to support cell growth can be significantly improved by suitable modification. PMID- 15332628 TI - Novel injectable urethral bulking agents for the treatment of urinary incontinence. AB - Stress urinary incontinence is a highly prevalent disorder resulting from weak urethral closure mechanisms. Endoscopic injection of a urethral bulking agent (UBA) under the urethral mucosa increases coaptation, which improves continence. Collagen is an efficient agent, although its effects are limited in time. Other materials still suffer either from a short-lasting effect or migration in distant organs. We evaluated here novel UBAs using an ex vivo model, with respect to criteria of ease of injection, ability to form a high and stable tissue bulking, implant elasticity and tissue reaction. One approach involves solutions of polymers in water-miscible organic solvents that precipitates in situ. In this manner, high and stable bulks were routinely obtained using various commercial polymers. Selected solvents reduced the tissue reaction to the implant. Microsphere suspensions in hydrogels also proved to be efficient UBA, although less stable bulks were obtained. Thermosetting chitosan hydrogels showed promising results with respect to bulk stability and isoelasticity with surrounding tissues. Different strategies have thus been compared and optimised ex vivo. Further experiments are required to compare the ability of these materials to induce a sustained in vivo bulking effect. PMID- 15332629 TI - Influence of beta-radiation sterilisation in properties of new chitosan/soybean protein isolate membranes for guided bone regeneration. AB - Novel chitosan (cts) and soybean protein isolate (SI) blended membranes were prepared. These membranes were produced by solvent casting. Besides combining the advantages of both materials, cts/SI membranes exhibit a biphasic structure that will eventually originate in situ porous formation, through a two-step degradation mechanism. In this particular work the effect of beta-radiation over the properties of these membranes was evaluated. beta-radiation sterilisation was performed at three different doses (25, 50 and 100 kGy) and eventual surface chemical changes were evaluated by Fourier transformed infrared--with attenuated total reflection and contact angle measurements. Moreover, eventual bulk properties changes due to beta-radiation were assessed by means of mechanical tensile tests and water uptake measurements. In general, no substantial changes were detected on the studied properties, with the exception of the surface energy that was found to be slightly increased for higher applied doses. PMID- 15332631 TI - Pre-mineralisation of starch/polycrapolactone bone tissue engineering scaffolds by a calcium-silicate-based process. AB - This work describes a new methodology to produce bioactive coatings on the surface of starch-based biodegradable polymers or other degradable polymeric biomaterials. As an alternative to the more typical bioactive glass precursors, a calcium silicate gel is being employed as a nucleating agent, for inducing the biomimetic formation of a calcium-phosphate (Ca-P) layer. The method has the advantage of being able to coat efficiently both compact materials and porous 3-D architectures aimed at being used on tissue replacement applications and as bone tissue engineering scaffolds. This treatment is also very effective in reducing the incubation periods, being possible to observe the formation of an apatite like layer, only after 12 h of immersion in a simulated body fluid (SBF). The apatite coatings formed on the compact surfaces or along the fibres of a fibre mesh scaffold structure made from a starch/polycrapolactone blend (SPCL) were analysed and compared in terms of morphology, chemical composition and structure. After the first days of SBF immersion, the apatite-like films exhibit the typical cauliflower like morphology. With increasing immersion times, these films exhibited a partially amorphous nature and the Ca/P ratios became very closer to the value attributed to hydroxyapatite (1.67). It was possible to fully pre mineralise the SPCL scaffolds and simultaneously to keep the porous morphology of the fibre-bonded scaffold. PMID- 15332630 TI - Investigation of microstructural features in regenerating bone using micro computed tomography. AB - We illustrate some of the uses of micro-computed tomography (micro-CT) to study tissue-engineered bone using a micro-CT facility for imaging and visualizing biomaterials in three dimensions (3-D). The micro-CT is capable of acquiring 3D X ray CT images made up of 2000(3) voxels on specimens up to 5 cm in extent with resolutions down to 2 microm. This allows the 3-D structure of tissue-engineered materials to be imaged across orders of magnitude in resolution. This capability is used to examine an explanted, tissue-engineered bone material based on a polycaprolactone scaffold and autologous bone marrow cells. Imaging of the tissue engineered bone at a scale of 1 cm and resolutions of 10 microm allows one to visualize the complex ingrowth of bone into the polymer scaffold. From a theoretical viewpoint the voxel data may also be used to calculate expected mechanical properties of the tissue-engineered implant. These observations illustrate the benefits of tomography over traditional techniques for the characterization of bone morphology and interconnectivity. As the method is nondestructive it can perform a complimentary role to current histomorphometric techniques. PMID- 15332632 TI - Continuous mandibular distraction osteogenesis using superelastic shape memory alloy (SMA). AB - Distraction osteogenesis is a well-established method of endogenous tissue engineering. It is a biological process of bone neo-formation between segments subjected to tension. The concept of this study was to investigate the distraction osteogenesis with a device capable of creating a permanent and constant force during the whole process as if a very large number of small elongations were applied constantly. The mechanical testing of the device used to produce the constant force and the in vivo analysis of the bone growth after it was implanted in rabbits are presented on this work. The device consists of a NiTi coil spring, superelastic at body temperature, in order to have a stress plateau during the austenitic retransformation during the unloading. The in vivo analysis was made on six female rabbits of 12 months old. A segmental mandibulectomy at the horizontal arm of the mandible and a corticotomy at 5mm distant from the gap were made. Next, following a latency period of five days, the SMA springs were implanted to induce the bone neo-formation. The displacement at the unloading plateau shows that it is necessary to have longer springs or to use several (available commercially) in series in order to fulfil the requirements of a human distraction. The temperature variations induced changes in the spring force. However, when the temperature returns to 37 degrees C the distraction force recovers near the initial level and does so completely when the distraction process continues. For the in vivo study, all six rabbits successfully completed the distraction. The radiographies showed the gap as distraction advanced. A continuity in the newly formed bone with similar transversal and horizontal dimensions than the original bone can be observed on the histologies. In conclusion, the application of a constant force on distraction osteogenesis, using SMA springs, may be a successful alternative to the conventional gradual distraction. PMID- 15332633 TI - [Therapeutic approach in locally advanced and complicated rectosigmoid and genital cancers]. AB - Locally advanced and complicated rectosigmoidian and genital cancers raise many therapeutic problems for surgeons. The most frequently used therapeutic methods nowadays are: radiotherapy, chemotherapy, surgical procedures, immunotherapy and other modern methods that aren't in the current clinical use yet. In a trial of 456 patients with locally advanced and complicated rectosigmoidian cancers and 632 patients with genital cancers we performed 573 (52.6%) radical surgical procedures and 515 (47.4%) palliative procedures, 301 (27.6%) of these being permanent colostomy (257 terminal and 44 in continuity). All of the patients received radiotherapy or chemotherapy pre and/or after surgery. The survival was between 5-7 months in the trial of patients with permanent colostomy, between 12 36 months in the trial of patients with palliative surgical procedures and adjuvant treatment and between 5-17 years in the trial of patients with radical surgical procedures and neo- and adjuvant therapy. PMID- 15332634 TI - [Locoregionally advanced rectosigmoid cancer: diagnosis, surgical approach, late results]. AB - The colorectal cancer continues to be diagnosed in advanced stages in our country, mainly due to unapplying of a programmer of active diagnosis through screening on the population with risk for colorectal cancer, and inefficiency of primary care system. In the department of General Surgery CFR Craiova Hospital between 1991-2001 were operated a number of 231 patients with colon cancer and 104 patients with rectal cancer. The results, showing an increased number of recurrences in cases of resection performed for advanced loco-regional tumors of rectosigmoid, made us to reconsider the attitude of avoiding the abdominoperineal resection even when the distance between the inferior limits of the tumors and the anal edge exceeded the distance considered being standard for a low anastomosis performing. The follow-up of the patients with paraclinic technique that didn't prove efficient led in the most cases to a delaying in diagnosis of local recurrences until the moment of resectability was exceeded. The applying of efficient methods in early diagnosis of colorectal cancer and follow-up could provide in the future better results for anterior resections with low anastomosis. PMID- 15332635 TI - [Cavitary viscera injuries during laparoscopy]. AB - Intraoperative incidents of laparoscopic surgery resulting in cavitary viscera injuries are analyzed, based on a series of 6308 patients (6800 operations) operated during a period of 9 years. This retrospective study includes 11 cases: 6 intraoperative cavitary viscera perforations (esophagus, gastric fornix, ileum, sigmoid) have been recognized and solved in the same laparoscopic operation or after conversion. In 5 cases the diagnosis was established in the postoperative period as fistulas (urinary bladder) or diffuse peritonitis caused by various types of injuries (esophagus, small and large bowel) and required reoperations. We present a global view of these cases and also analyze the determinant causes of the complications, emphasize the importance of intraoperative identification of such injuries and specify the place of minimally invasive surgery in the treatment of such accidents with an optimum healing and low rate of postoperative morbidity and mortality. PMID- 15332636 TI - [Cholelithiasis after gastric surgery--possibilities and limits of laparoscopic approach]. AB - An increased incidence of cholelithiasis has been widely reported after gastric surgery. In the early phase of patient selection, previous gastric surgery has been considered a relative contraindication to laparoscopic cholecystectomy. Between 1999-2003, in our clinic, laparoscopic cholecystectomy was attempted on 3145 patients. Of these patients, 37 patients had gallstone, which occurred several years (mean 17.9 years) after gastric surgery for peptic ulcer (34 cases), gastric cancer (1 case) and hiatal hernia (2 cases). Surgical procedures that had been performed included Billroth I gastrectomy (14 cases), Billroth II gastrectomy (15 cases), truncal vagotomy and piloroplasty (6 cases), and Nissen (2 cases). On this group there were 19 chronic cholecystites, 17 acute cholecystites and one cholesterolosis with gallbladder polyps. One patient was diagnosed with gallstone in common bile duct, successfully removed preoperatively after endoscopic sphincterotomy. The laparoscopic approach was possible in 26 cases. The technical difficulty after Cuschieri was level III--26 patients and level IV--11 patients (reconversion). The mean operative time was 75 minutes. The postoperative evolution of the patients was good in 36 cases. One case had a biliary fistula. The mean postoperative hospital stay was 3.6 days for laparoscopic cholecystectomy and 9 days after conversion. The postoperative adhesions determine the main technical difficulty, but it proved to be a relative contraindication for laparoscopic cholecystectomy for gallstones after gastric surgery. Clear visualization of anatomic structures and landmarks, and scrupulous hemostasis are needed to perform a safe laparoscopic cholecystectomy in these patients. PMID- 15332637 TI - [The importance of forseeing and early diagnosis of thromboembolic disease]. AB - The aim of this study is to assess the incidence of thromboembolism in laparoscopic cholecystectomies. 100 unselected patients undergoing laparoscopic procedures performed by the same team were studied. All patients received preoperative prophylaxis with low/molecular weight heparin (LMWH), which was continued until full mobility. Four cases of deep venous thrombosis of lower limbs were clinically identified and confirmed by means of Doppler ultrasound examination. There were no cases of pulmonary embolus. Deep venous thrombosis occurred during the prophylactic administration of LMWH in the fourth postoperative day, consequently intravenous treatment with Heparin and then with oral anticoagulants was required. In each observation the operation took more than one hour. Among the patient-dependent risk factors, we have identified: age above 40 years, obesity, history of deep venous thrombosis, localized preoperative infection, congestive cardiac failure. Although the thromboprophylaxis has been performed within the laparoscopic surgery similar to that recommended in the classical procedures, when a high risk has developed the illness started. We underline the importance of a careful postoperative clinical monitoring in order to prevent the serious accidents that may appear. PMID- 15332638 TI - [Laparotomy in the treatment of severe intraperitoneal infections]. AB - A retrospective clinical study was carried out on a group of 18 patients with severe intraperitoneal infections due to various causes. The decision to use laparotomy was supported by the severity of sepsis (APACHE II score varied from 14 to 30), highly septic peritoneal fluid, the features of peritonitis, patients with impaired immunity. Open packing was realized with a Dacron mesh sutured to the fascial margins, associated with multiple peritoneal drainage in all cases. Postoperative mortality was 50%, mainly to rapid evolution of septic shock, unresponsive to intensive care support. Secondary suture was performed after granulation of the wound, on the day 14-21 postoperatively followed at 3-4 months by definitive repair of the abdominal wall in 2 cases. Laparotomy proved to be an efficient treatment in severe peritonitis, that should be used prior to irreversible phases of septic shock. PMID- 15332639 TI - [Multivisceral operations for carcinoma of the upper stomach and cardia]. AB - In this study are noted technical problems regarding "en bloc" multiple organ resections and the anatomic and functional reconstruction for carcinoma of the upper stomach and cardia. From 1997 to 2002, a total of 264 patients with cancers of the stomach were operated in the service. 75 patients presented cancers localized at the proximal stomach and cardia (97.33% adenocc.). The rate of resectability was 27.77% (27 pt.). Types of operations in this series were: standard esophagogastrectomy in 7 patients; total gastrectomy with regional lymphadenectomy in 9 patients; 11 patients underwent "en bloc" multiple organ resection, with the removal of the stomach, partial or total esophagectomy and, occasionally, ablation of the spleen, pancreas, left hepatectomy, resection of the diaphragm and an extensive lymphadenectomy. Surgical mortality for the complex multivisceral resections was noted in 3 patients (8.88%). The global 5 years survival in the service is poor: 15.9%. PMID- 15332640 TI - [Particular evolution of a jejunal tumor in one patient with multiples neoplasia]. AB - The authors present the case of one male patient, 71 years old, known with serial neoplasia (right colon neoplasm 1983, left colon neoplasm 1987) for which there were performed several interventions. The patient was admitted first time in our Department in 1998 for a non specifically symptomatology; in particular anaemic syndrome. The first intervention was performed in June 1998, without finding anything pathological, despite the presumption of a third tumor on the remaining colonic territory, theory that was partially sustained by the paraclinical examinations. The second intervention was performed in February 2001, when it was found and extirpated a jejunal tumor, under Treitz angle, but after 9 months, when a new intervention was performed, we found a relapse tumor which was not extirpable. The aim of this study is to present the unpredictable versatility of small bowels tumors and the difficulty of establishing an early positive diagnose. PMID- 15332641 TI - Subclavian vein flexible guidewire knotting. A potential serious complication in hemodialysis patients. AB - BACKGROUND: Complications after subclavian vein catheterization are well documented in the literature. The purpose of this article is to present a case of a rare and potential serious complication of flexible guidewire knotting in subclavian vein catheterization in a hemodialysis patient. METHOD: A 72 year-old woman on hemodialysis due to diabetes has had left upper extremity arteriovenous fistula (AV-F) thrombosed on the scheduled hemodialysis day. A right subclavian vein catheterization for hemodialysis was decided and attempted, that ended up with a flexible guidewire knotting. RESULTS: The flexible guidewire was splinted with the Kit's dilator and after great effort, under radioscopic control the knotting was managed to be unknotted and the guidewire removed. CONCLUSIONS: Our case suggests that a rare and potential serious complication of subclavian vein catheterization for hemodialysis can be successfully managed with appropriate approach and skillful maneuvers. PMID- 15332642 TI - [Clinical and therapeutic considerations in a case of colonic carcinoma in a child]. PMID- 15332643 TI - [Retroperitoneal tumor]. AB - This paper presents the case of a 69 year old patient who has a benign retroperitoneal tumor (lipofibroma) presenting differential diagnosis problems (in special preoperative one, and intraoperative technique difficulties, revealing the fact that, in this case, paraclinical investigations have an orientative value, without offering etiologic information, and do not allow a clear preoperative strategy. Tumor resection was performed under an intraoperative histologically diagnosis of lipofibroma of the retroperitoneal region. Postoperative evolution was good. PMID- 15332644 TI - [Protective transanal tube in colo-rectal anastomosis]. AB - A particular issue of colo-rectal resection is represented by the great frequency of anastomotic dehiscences, which the medical literature has shown to vary between 3.25% and 30%. This justifies the surgical interest in perfecting methods aimed at reducing the frequency of this complication. One of these methods is the protective ileo- or colostomy, which has successfully reduced the frequency of dehiscences down to 4%. To the same propose a method that has been documented in previous literature and which consists of inserting an endoanal protective tube, above the anastomosis. The tube is connected to a plastic pouch and is maintained for 3-6 days, until transit has resumed. By applying this method to 38 cases, we noted only two cases (5.2%) of anastomotic leakage with small output, which were solved spontaneously after a few days. The basic nature and efficiency of this method have lead us to propose it as an alternative procedure to protective ileo- or colostomy. This method has the advantage of preventing further procedures. The quality of life of these patients is appreciably improved. PMID- 15332645 TI - [Peroral cholangioscopy]. AB - The aim of this study was to present peroral cholangioscopic method, their technique, indications, contraindications, the advantages and disadvantages of this method. Peroral cholangioscopy is associated with ERCP and was developed in the last two decades by development of "mother & baby" endoscopic system. In same mode with percutaneous cholangioscopy or laparoscopic cholangioscopy, peroral cholangioscopy have an important role in diagnosis and therapy of common bile duct diseases. In 2001-2002 the authors have perform six peroral cholangioscopy for diagnosis and therapy (nonspecific radiological findings on ERCP) and in five cases, they have found choledochal lithiasis which was resolved by this way. In one case has not found any pathology. By their efficiency with a low rate of specific complications, peroral cholangioscopy is now an appropriate method in diagnosis and therapy of common bile duct disease. PMID- 15332646 TI - [The extraperitoneal transomphalic drainage of the postoperative hydatid cavity through the transposed round ligament detached from the liver]. AB - The transomphalic and then the transligamentary extraperitoneal drainage imagined by D. Burlui and its multiple use in the liver and the biliary tract surgery, is well known. Concerning the hepatic hydatid cysts, the drainage of the remaining cavity by the round ligament way is possible for most of the locations of the parasite, but it is less performed in the right liver lateral segments locations, too far situated, and also in the left liver locations. On the other hand, this specific method implies to place the drainage tube using the round ligament "in situ ", normally placed and inserted. We encountered an extremely large hydatid cyst of the left hepatic lobe, adherent to the anterior and posterior abdominal wall, spleen, stomach, pancreas and large bowel, which required to extend the incision to the left and make the necessary segmentation of the round ligament at its hepatic insertion. After pericystotomy and the evacuation of the cyst, the remaining cavity drainage was performed through the same round ligament whose free end was tightly fixed to the partial pericystotomy border. This transposition allowed the exteriorisation of the drainage tube in the same way as for the transligamentary and total extraperitoneal original method. PMID- 15332647 TI - Development of coincidence transmission electron microscope. III. Incorporation with gamma-type imaging energy filter. AB - We have developed a new analytical transmission electron microscope (TEM), called coincidence TEM, which, in principle, enables observation of elemental mapping images at a high signal-to-noise ratio. We have previously reported the successful observation of an elemental mapping image of a specimen, but over a very long period of time (168 h). To solve this inefficiency, we installed a gamma-type imaging energy filter in the coincidence TEM to remove the no-loss electrons, which are mainly transmitted electrons. This has enabled the intensity of the background signals in the coincidence measurement to be markedly reduced. The coincidence TEM with a gamma-type imaging energy filter allows the coincidence image to be observed in 3 h, thus, the measurement time is shortened by two orders of magnitude. Moreover, the use of a silicon drift detector (SDD) will shorten the measurement time. PMID- 15332648 TI - Observation of iron silicide formation by plan-view transmission electron microscopy. AB - The formation and the phase transitions of iron silicide by solid-phase epitaxy have been investigated by means of plan-view transmission electron microscopy, which enables us to observe a clean interface between Fe and Si. Layers of Fe were deposited on Si (100) at room temperature in an ultrahigh vacuum chamber. The sample was annealed in the electron microscope at a temperature between 673 and 1073 K. After annealing at 673 K, FeSi crystallites were formed with various orientations. When the annealing temperature was increased to 973 K, we found that the crystallites suddenly started to coalesce into grains of several hundreds of nanometers in size and polycrystalline beta-FeSi2 was formed. These phase transitions were also confirmed with electron energy-loss spectroscopy. PMID- 15332649 TI - Monte Carlo simulation of topographic contrast in scanning ion microscope. AB - Topographic contrast of secondary-electron (SE) images in a scanning ion microscope (SIM) using a focused gallium (Ga) ion beam is investigated by Monte Carlo simulation. The SE yield of heavy materials, in particular, due to the impact of 30 keV Ga ions increases much faster than for the impact of electrons at < or =10 keV as a function of the angle of incidence of the primary beam. This indicates the topographic contrast for heavy materials is clearer in a SIM image than in a scanning electron microscope (SEM) image; for light materials both contrasts are similar to each other. Semicircular rods with different radii and steps with large heights and a small wall angle, made of Si and Au, are modeled for comparison with SE images in SEM. Line profiles of the SE intensity and pseudo-images constructed from the profiles reveal some differences of the topographic contrast between SIM and SEM. We discuss not only the incident-angle effect on the contrast, but also the effects of re-entrances of primary particles and SEs to the neighboring surface, the effect of a sharp edge on the sample surface, and the effects of pattern size and beam size. PMID- 15332650 TI - The effect of the signal-to-noise ratio in CBED patterns on the accuracy of lattice parameter determination. AB - A method of higher-order Laue zone line position measurement in convergent-beam electron diffraction (CBED) is proposed based on Hough transformation. A thorough analysis of the errors introduced by this measurement procedure is performed and their influence on the accuracy of lattice parameter determination is estimated. A criterion is derived which enables the accuracy to be predicted before experimental measurements are made and, thus, allows the selection of the best CBED geometry for parameter measurement. PMID- 15332651 TI - Contrast-to-gradient method for the evaluation of image resolution taking account of random noise in scanning electron microscopy. AB - We have taken random image noise into consideration in the contrast-to-gradient (CG) method for the evaluation of image resolution R in scanning electron microscopy (SEM). When looking at the local fine pattern in the SEM micrograph containing much random noise, viewers gradually expand the region of interest (ROI) to improve the signal-to-noise ratio (SNR) and to recognize the pattern. We employed this approach in the CG algorithm to evaluate potential resolution Rpot, which is defined as the minimal/most accurate CG resolution calculated in the ROI expanding process. The image noise or SNR also is evaluated by the parameter deltaR / R, where deltaR is a standard deviation of R. The Rpot values depending on SNR are useful for comparison among images containing different random noise. PMID- 15332652 TI - HAADF-STEM imaging with sub-angstrom probes: a full Bloch wave analysis. AB - A full coherent Bloch wave calculation is presented to investigate high-angle annular dark-field image formation for sub-angstrom probes in scanning transmission electron microscopy (STEM). With increasing illumination angle, the contribution of the 1s bound state increases to a maximum at an optimum probe angle, after which we find increasing contributions from high-angle plane wave states around the periphery of the objective aperture. Examination of image contributions from different depths within a crystal shows an oscillatory behavior due to the beating between 1s and non-1s states. The oscillation period reduces with decreasing probe size, while the relative contribution from a specific depth increases. This signifies a changeover from a projection mode of imaging to a depth-slicing mode of imaging. This new mode appears capable of resolving three-dimensional atomic structures in future generation aberration corrected STEM. PMID- 15332653 TI - Analytical electron microscopy and electron holography on microstructures and magnetic domain structures of Sm-Co 2:17 magnets. AB - Microstructures and magnetic domain structures of precipitation-hardened Sm-Co permanent magnets were systematically investigated by analytical electron microscopy and electron holography. By an elemental mapping method with energy dispersive X-ray spectroscopy, the change in the local distribution of additive elements, i.e. Cu, Fe and Zr, in Sm-Co magnets with various heat treatments was visualized and the enrichment of Zr in the Z-phase with a width of approximately 1 nm was clarified directly. Detailed analysis with electron holography revealed that considerable fluctuation in the distribution of lines of magnetic flux in the step-aged magnet was due to the chemical partitioning of additives and resulted in magnetic hardening during the magnetization process. PMID- 15332654 TI - Practical method to determine the filter shape function used in the three dimensional Fourier filtering method. AB - The procedure to determine the appropriate filter shape function used in the three-dimensional Fourier filtering method (3D-FFM) is discussed from a practical point of view, so as to reduce the artificial contrast induced by the processing and to achieve a high signal-to-noise ratio. The effects of cutting off the high spatial frequency of the filter shape function and damping at low spatial frequency were investigated by using through-focus images of a gold (110) thin film. In addition, the effect of the width of the filtered area was discussed using through-focus images of a carbon nanotube. For reliable image processing, the filter shape function should be cut off beyond the information limit and attenuated with damping at low spatial frequency. Furthermore, the extraction area should include the distributed area of the relevant structural components appearing in the 3-D Fourier spectrum. PMID- 15332655 TI - Chemical junction delineation of a specific site in Si devices. PMID- 15332656 TI - Ontogenesis of peripheral electromagnetic receptors in hornets. AB - This article traces the ontogenesis of peripheral electromagnetic receptors (PER) in the cuticle of the Oriental hornet (Vespa orientalis). In the abdominal cuticle of adult hornets, the PERs are densely distributed throughout, but there are even more than 30 at the margins of the segments. These organelles develop as a network in the hornet cuticle immediately upon its completion. Briefly, from each basic cell of a PER grows a bulge towards the exterior, that is, towards the illuminated region of the cuticle. This bulge develops rapidly and as it grows it starts to push out and lift up the various layers of the cuticle, the while pressing them together. By a spiraling movement, the bulge insinuates itself between the layers, whereupon it dissolves and punctures its way through all the layers of the hypocuticle, via the endocuticle up to the exocuticle. The only cuticular layer that remains intact is the epicuticle, but even that undergoes change, assuming the shape of a smooth surface with a depression at its center. The indented part in the epicuticle is circular, approximately 2.5 microm in diameter and enables the entry of radiation (illumination) from the outside into the PER, which is located half-way down the cuticle, with the distance from the exterior to the base of the PER being approximately 25 microm. The numerous lamellae of the cuticle run parallel to one another, but in the region of the bulge they are either perpendicular or directed upwards. This ontogeny of the PERs lends the cuticle a sandwich-like shape, being radically perforated by the PERs bulges, yet covered at the top by the epicuticle and at the bottom by basal cells. The PERs also extend shoots into the cuticular layer and these further perforate the cuticle but also interlink the various PERs. From all the above, it is clear that the cuticle forms first and only subsequently does the network of PERs develop and interpenetrate its various layers. PMID- 15332657 TI - Silk structure in the hornet cocoon. AB - This study deals with the development of a silk coat around the growing larva of the Oriental hornet (Vespa orientalis). Prior to its metamorphosis into a pupa, the larva secretes a silk weave, which enwraps it completely. This silk coat is not uniform throughout, but rather varies along the extent of the larval body. First, it is possible to discern in the spun silk weave fibers whose thickness is approximately 10 microm; in which case such a fiber is actually a duplet of two individual fibers, each with its own silk coat and an inner fibril (core). Additionally, there are silk plaque-like surfaces of a greater diameter and of variable shape, depending on the contour that they cover. In the cap region, i.e. the region where the head of the larva (and later of the developing imago) is located, the fibers are quite numerous and arranged in many (up to 10) layers, while the silk plaques are small. For the remainder of its body, the pupating larva produces a silk sleeve in which there are few fibers with numerous plaques in between, occupying most of the area. The larval silk fibers proper are of fairly uniform diameter. Apart from the pupating larvae, the adult hornets also secrete a type of silk, which acts as a glue holding together the components of the larval cell wall and their sticky silk is of variable thickness, depending on the configuration of the cell. The temperature of the silk varies in accordance with age of the pupating larva. Thus, the closer the developing hornet to eclosion, the lower the temperature. The present study reviews and summarizes this transformation process in the silk weave from its inception until eclosion of the imago. PMID- 15332658 TI - Elasticity of flagellar hooks. PMID- 15332659 TI - Relationship of sediment redox conditions to methyl mercury in surface sediment of Louisiana Lakes. AB - Surface sediment from three Louisiana Lakes containing overlying water layer spiked with 2 microg/g (2 ppm) mercury were incubated under oxygenated (air) and nonoxygenated (N2) conditions for determining the impact of oxygen status of overlying water on methylation of Hg in surface sediment from these lakes. The added mercury resulted in a greater than ten fold increase in methyl mercury (MeHg) as compared to native concentration of MeHg. The increase in methyl Hg production was less in sediment in which overlying water was exposed to oxygen rather than nitrogen. Results suggest that methyl Hg production would be less in lakes containing an oxygenated water column. In parallel microcosm studies without added mercury. MeHg decreased in sediment when redox potential of sediment suspension was increased from -200mV to +50mV. Results of these studies demonstrate the importance of oxygenation or redox condition of surface sediment on mercury methylation and demethylation. Sediment conditions, which either reduce methylation or enhance demethylation in surface sediment, will limit the bioavailability of MeHg to the aquatic environment. PMID- 15332660 TI - Development of a two-dimensional analytical model for predicting toxic sediment plumes due to environmental dredging operations. AB - A two-dimensional analytical transport model is developed to predict the dredge induced plume concentration in the horizontal plane under steady-state for simple hydraulic conditions. The derivation of the analytical solution is based on the solution to the advection-diffusion equation. The application of this analytical model is limited to mechanical dredge operations, bucket dredge, which has continuous point source in the water column. The analytical model developed herein is used to simulate sediment concentration distributions from a hypothetical dredging site and compared to results from a similar model using Stokes' settling. The result shows that Stokes' settling underestimates settling in the near-field and overestimates settling in the far-field plume. In summary, the model provides the ability to account for the rapid settling of larger particles in the suspended sediment near the dredging operations while simulating the slower settling and diffusive transport of smaller particles moving away from the dredging sites. PMID- 15332661 TI - Adsorption-desorption characteristics of lead in variable charge soils. AB - Adsorption desorption processes of Pb at contaminated levels in two variable charge soils were investigated. The red soil (RAR) developed on the Arenaceous rock (clayey, mixed siliceous thermic typic Dystrochrept) adsorbed more Pb2+ than the red soil (REQ) derived from the Quaternary red earths (clayey, kaolinitic thermic plinthite Aquult). The maximum adsorption values (Xm) that were obtained from the simple Langmuir model were 52.6 mmol Pb2+ kg(-1) soil and 29.9 mmol Pb2+ kg(-1) soil, respectively, for the RAR and REQ. Adsorption of Pb2+ decreased soil pH by 1.10 unit for the RAR soil and 1.21 unit for the REQ soil at the highest loading. The adsorption equilibrium pH of RAR was higher than that of REQ at the same Pb2+ concentration. The distribution coefficient (Kd) of Pb in the soils decreased exponentially with increasing Pb2+ loading. Most of the adsorbed Pb2+ in the soils was not desorbed in the 0.01 mol L(-1) NaNO3 solution. After five successive extractions with NaNO3, only 0-11% of the total adsorbed Pb2+ in the RAR soil was desorbed and the corresponding value of the REQ soil was 0-19%, indicating that the RAR soil had a greater affinity for Pb2+ than the REQ soil at the same Pb2+ loading. Different mechanisms might be involved in Pb2+ adsorption/desorption at different levels of Pb2+ loading and between the two soils. PMID- 15332662 TI - Determination of nonsteroidal anti-inflammatory drugs, caffeine, and triclosan in wastewater by gas chromatography-mass spectrometry. AB - Pharmaceuticals are a class of chemicals whose fate in the environment has received increasing attention in the past few years. A quantitative method was developed for the determination of acidic pharmaceuticals (ibuprofen, naproxen, ketoprofen, mefenamic acid, and diclofenac), caffeine and the antibacterial triclosan in wastewater effluent. The compounds were extracted from wastewater samples on Waters Oasis HLB solid-phase extraction columns, derivatized with N,O bis [Trimethylsilyl] trifluoroacetamide (BSTFA) and analyzed using gas chromatography-mass spectrometry. Estimated method detection limits ranged from 6 to 45 ng/L based on replicate analyses (n = 10). This method was applied to the analysis of effluent from a wastewater treatment plant and compounds were detected at concentrations of 18-72 ng/L. PMID- 15332663 TI - Determination of arsenic in water samples treated with ozone. AB - The applicability of ozone as a digesting agent in the trace analysis of arsenic in natural water samples has been investigated. Differential pulse polarographic (DPP) and spectrophotometric (SP) methods were applied for the determination of arsenic in ozone and, as reference, in persulphate treated samples. Additionally, inductively coupled plasma mass spectrometry (ICP-MS) was used for the total arsenic determinations. No significant differences were found between the results of the measurements using different techniques in the investigated natural ground water samples. Similar detection limits of 5 microg/L, lower quantification limits of approximately 10 microg/L and linearity limit up to 300 microg/L were estimated for the both DPP and SP measurements. Thus, ozone treatment may, in many cases, be successfully used for the pretreatment of samples prior to both DPP and SP determination of arsenic in natural water samples. PMID- 15332664 TI - A review of the occurrence and fate of naphthenic acids in aquatic environments. AB - Naphthenic acids are comprised of a large collection of saturated aliphatic and alicyclic carboxylic acids found in hydrocarbon deposits (petroleum, oil sands bitumen, and crude oils). Naphthenic acids enter surface water systems primarily through effluent discharge, but also through groundwater mixing and erosion of riverbank oil deposits. Of the possible environmental receptors (i.e., air, soil, and water), the most significant is water. Ambient levels of naphthenic acids in northern Alberta rivers in the Athabasca Oil Sands are generally below 1 mg L( 1). However, tailings pond waters may contain as high as 110 mg L(-1). The complexity of natural naphthenic acids in petroleum deposits poses an analytical challenge as reflected by the several techniques reported for quantitation of naphthenic acids in the environment. Although naphthenic acids are known to be persistent biomarkers used in identification of oil source maturation, little is established regarding their relative degradation pathways in aquatic environments. Published research related to the potential for microbiological degradation and adsorption to typical Athabasca Oil Sands soils reveal that naphthenic acids are likely to persist in the water column and, with prolonged exposure, accumulate in sediments. However, other than a very general knowledge of environmental persistence, the occurrence and fate of naphthenic acids has been sparsely studied. This article brings together some of those environmental persistence results, as well as detailed information regarding the origin of naphthenic acids in tailings ponds, chemistry and toxicological considerations, current analytical methods for aquatic sampling, and areas of future remediation research. PMID- 15332665 TI - By-side impurities in chloronaphthalene mixtures of the Halowax series: all 12 chlorobenzenes. AB - Total chlorobenzene content of the Halowax mixtures ranged from 1100 to 9800 ng/g. A lower chlorinated Halowax 1031 and 1000 were a several times more contaminated with chlorobenzenes than higher chlorinated Halowax 1001, 1099, 1013, 1014, or 1051. Depending on the type of the Halowax mixture the most contributing amongst of chlorobenzene homologue groups were di-, tetra-, penta-, and hexa-CBz. A dominance and specific profile of 1,4-DiCBz, PeCBz, and HCBz were characteristic to Halowax 1001, 1099, 1013, 1014, and 1051, while for other formulation the CBzs profile varied, and 1,2,3,4-TeCBz (Halowax 1031) and 1,4 DiCBz (Halowax 1000) predominated. A steric hindrance effect seems to direct a preferential by-side formation of 1,4-DiCBz and further also of PeCBz and HCBz due to relatively elevated temperature as well as duration time of synthesis for five most chlorinated (49-70 Cl%) chloronaphthalene Halowax formulations. PMID- 15332666 TI - By-side impurities in chloronaphthalene mixtures of the Halowax series: all 19 chlorophenols. AB - All seven Halowax formulations examined were contaminated with chlorophenols (CPhs). The total CPhs content of the CN mixtures ranged from 1050 to 34,200 ng/g. The absolute concentration of CPhs decreased with an increase of degree of chlorination (% Cl) of the Halowax 1031 (27%), 1000 (35%), 1001 (49%), and 1099 (51%), respectively, but remained relatively constant for higher chlorinated Halowax 1099 (51%), 1013 (54%), 1014 (59%), and 1051 (70%). 2,4,6-TrCPh and 2,4 DiCPh did contaminated all seven Halowax formulations and PeCPh remained undetected (<90 ng/g) only in Halowax 1099, while less frequently occurred 2,6 DiCPh and followed by 3-/4-MoCPh, 2,5-DiCPh, 2-MoCPh, 2,3-DiCPh, and 2,3,4,6 TeCPh. The CPh congeners such as 3,4-DiCPh, 3,5-DiCPh, 2,3,4-TrCPh, 2,3,5-TrCPh, 2,3,6-TrCPh, 2,4,5-TrCPh, 3,4,5-TrCPh, 2,3,4,5-TeCPh, and 2,3,5,6-TeCPh were undetected in any of the Halowax formulation examined. The profile (%) of higher chlorinated CPh homologues like PeCPh in the Halowaxes followed somehow a degree of chlorination (Cl %) of the parent mixture, i.e., increased from 14, 22, 43, 55, 52 to 80%, respectively, but exception was free of PeCPh the Halowax 1099. And contrary, for a lower chlorinated DiCPhs and TrCPhs their relative proportions decreased but again an exception was Halowax 1099. The fingerprint profiles of 2,4,6-TrCPh, 2,4-DiCPh, and PeCPh, which were usually the most abundant congeners varied and were less consisted when compared to the CPh homologue group profiles, and also more depended on the type of technical Halowax mixture. Due to absence of 2,4,5-TrCPh but presence of 2,4-DiCPh, 2,6-DiCPh, 2,4,6-TrCPh, and PeCPh, which, respectively, are indicative congeners for CPhs obtained by alkaline hydrolysis of chlorobenzenes or phenol chlorination using gaseous chlorine, just co-occurrence of phenol impurity in technical naphthalene is suggested as a solely source of CPhs in the Halowax mixtures. PMID- 15332667 TI - By-side impurities in chloronaphthalene mixtures of the Halowax series: all 209 chlorobiphenyls. AB - 202 congeners of chlorobiphenyl (CB) with 109 single-resolved compounds and 93 co eluting, which represented from mono- to deca-CB were quantified as by-side impurities in all seven and of various type the technical chloronaphthalene (CN) formulations of the Halowax series. IUPAC No. 104, 145, 159, 169, 184, 186, and 188 were undetected (<0.05-<0.7 ng/g). The total CBs content of the Halowax formulations ranged between 220 and 640,000 ng/g, and lower chlorinated Halowaxes 1031 (27% Cl) and 1000 (35% Cl) were two to three orders of magnitude more contaminated with CBs than higher chlorinated Halowax 1001, 1099, 1013, 1014, and 1051 (49-70% Cl). The profile of CB homologue groups of the Halowaxes changed and followed somehow a degree of chlorination of the parent CN mixture. For Halowax 1031 and 1000, a decreasing trend in proportion from mono- to deca-CB was evident, while for Halowax 1001, 1099, 1013, 1014, and 1051 the relative proportions between the CB homologue groups varied somehow. Evidently content of lower chlorinated mono- and di-CBs highly decreased and steadily increased tri- to octa-CBs and they become dominating homologues in Halowax formulations in parallel with an increase of the content of higher molecular weight CN constituents, respectively. Among planar non- and mono-ortho CBs, total concentrations in Halowax formulations were detected in the range of 2.0-2600 ng/g, and only 3,3',4,4',5,5'-HxCB (no. 169) was undetected (<0.05ng/g). Detection of by-side CBs in technical Halowaxes demonstrate clearly that those formulations apart from a massive introduction of dioxin-like CNs become in the past also an early source of environmental pollution with CBs, which proceeded for around 20-30 years use of original CB formulations. PMID- 15332668 TI - Treatment and biodegradation kinetics of microbially treated domestic wastewater sludge. AB - A laboratory-scale study was undertaken to evaluate the liquid state bioconversion (LSB) in terms of biodegradation of microbially treated domestic wastewater sludge (biosolids) as well as its kinetics. The potential fungal strains and process factors developed from previous studies were used throughout the study. The results presented in this study showed that an effective biodegradation occurred with the biosolids (sludge cake) accumulated. The maximum biosolids (sludge cake) accumulated (93.8 g/kg of liquid sludge) enriched with the biomass protein (30.2 g/kg of dry biosolids), was achieved which improved the effluent quality by enhancing the removal of chemical oxygen demand (COD), reducing sugar (RS), soluble protein (SP), total dissolved solids (TDS), and total suspended solids (TSS). The higher reduction of specific resistance to filtration (SRF) was observed during bioconversion process. The kinetics results showed that the experimental data were better fitted for the biodegradation efficiency, and biosolids accumulation and biodegradation rate. PMID- 15332669 TI - Contribution of cell outer membrane and inner membrane to Cu2+ adsorption by cell envelope of Pseudomonas putida 5-x. AB - The role of outer and inner membrane in the Cu2+ adsorption process by gram negative bacterium Pseudomonas putida 5-x, which was isolated from local electroplating effluent with high Cu2+ accumulating capability, was studied. The results indicate that both the outer and inner membrane exhibited high Cu2+ adsorption capacity. Outer and inner membrane contributed about 30-35% and 20-25% parts of adsorption capacity in Cu2+ adsorption by cell envelope of Pseudomonas putida 5-x, respectively. The total contribution of outer and inner membrane to Cu2+ adsorption by cell envelope was much greater than that of peptidoglycan layer. The relatively high phospholipid content in the outer membrane might result in its greater heavy metal ions adsorption capacity. The Cu2+ binding process by the outer and inner membrane of Pseudomonas putida 5-x is the adsorption processes and can be described with Freundlich isotherms. PMID- 15332670 TI - Regrowth evaluation of coliform bacteria injured by low chlorine doses using selective and nonselective media. AB - In this study, the repairing capacity of coliform bacteria injured by chlorine is determined. Chlorine doses from 0.014 to 0.070mg L(-1) were used in according to frequency (up to 38%) of chlorine concentrations detected in a drinking water distribution network, adopted as case study. m-Endo (selective) and m-T7 (nonselective) cultivation media are used. Bacterial regrowth was detected up to 0.035 mgL(-1) of chlorine for 4h of incubation. The coliform bacteria were not able to regrowth when the dose of chlorine increased to 0.07 mgL(-1). Bacterial regrowth increased by increasing C:N ratio from 1:40 to 11:40. m-T7 (nonselective) medium allowed to detect bacterial regrowth also for lower incubation periods. Chlorine doses higher than 0.2 mg L(-1) dose at water source with a low total organic carbon (TOC) content are recommended to control bacterial regrowth in the distribution network. PMID- 15332671 TI - Biodegradation of phenol by acclimatized Pseudomonas putida cells using glucose as an added growth substrate. AB - Biodegradation of phenol, a pollutant derived from many industrial processes, was achieved through acclimatized Pseudomonas putida cells. The strategy to overcome the inhibitory effect of phenol on microbial growth involved the addition of glucose, a conventional carbon source. A factorial experimental design was employed in order to optimize the initial phenol and glucose concentrations. The optimum conditions found were applied in 2-lt bioreactors. The development of acclimatized cells and the use of glucose as an added growth substrate resulted in a significant phenol degradation rate of 60.7 mg L(-1) h(-1) with a complete removal of 1200 mg L(-1) phenol. PMID- 15332672 TI - Concentration of selected metals in liver, kidney, and muscle of the red deer (Cervus elaphus). AB - Concentration of cadmium, lead, chromium, zinc, copper, and manganese in liver, kidney, and muscle of red deer was investigated. For analysis of the content of these trace elements an AAS method was used. The concentration of cadmium was significantly (p < 0.05) higher in kidney in comparison with liver and muscle. The level of lead was the highest in muscle (p < 0.05). Lower values were detected in liver and in kidney. The concentration of chromium was very similar in all studied tissues. A higher concentration of zinc was found in muscle, followed by kidney and liver. The level of copper was significantly higher in liver (p < 0.05), and lower in kidneys and muscle. In evaluation of the concentration of manganese in red deer, the highest concentration of this element in liver was detected. PMID- 15332673 TI - Effect of coupled semiconductor system treating aqueous 4-nitrophenol. AB - This article describes a method for treating aqueous 4-nitrophenol by mixing with TiO2-SnO2 coupled particles. The reactivity of single photocatalysis (TiO2 and SnO2 all as 1.2 g/L) is compared with that of mixing TiO2-SnO2 (0.6 g/L + 0.6 g/L) coupled particles. Aqueous 4-nitrophenol was treated by single photocatalysis or coupled semiconducting particles in a double-layer glass batch reactor using a 15W UV fluorescent tube at 25 degrees C and 300 rpm for mixing. The results demonstrated that pH and the concentration of TiO2 were the factors that most influenced the degradation characteristics, and that the system of TiO2 SnO2 coupled particles improved the efficiency of removal of refractory organic pollutants by 15%, by the inter-particle electron transfer (IPET) effect. The results showed that the degradation of aqueous 4-nitrophenol was 75% when coupled particles were used--better than the 60% obtained using single photocatalysis- with a reaction time of 120min. PMID- 15332674 TI - Pilot scale microfiltration-coagulation for treatment of retention pond water. AB - An evaluation of two commonly used coagulants, alum and ferric chloride was conducted to treat retention pond water using microfiltration. To determine the effectiveness of these coagulants in removing turbidity, color, and total suspended solids two different sets of the experiments were performed. Preliminary test was carried out to evaluate the optimum dosages of coagulants. Optimum turbidity removal was achieved with a 4 and 20 mg/L dosage for ferric chloride and alum, respectively. Generally, coupling microfiltration with coagulation using both alum and ferric chloride exhibited excellent effectiveness for turbidity, color, and total suspended solids removal. The efficiency for alum and ferric chloride for turbidity removal were 96 and 98%, respectively, which was greater than 89% removal using microfiltration alone. Furthermore, microfiltration only demonstrated 81 and 83% removal efficiency for color and total suspended solids removal, respectively. However, microfiltration coagulation using alum and ferric chloride resulted about 83 and 93% color removal, and 92 and 94% total suspended solids removal, respectively. PMID- 15332675 TI - Pore size and adsorptive capacity of unburned carbon affected by gasification with carbon dioxide. AB - Gasification of unburned carbon by using CO2 gas at temperatures of 950-1100 degrees C was performed, in order to modify the pore size distribution and adsorptive capacity of the carbon. Equipments such as N2 adsorption apparatus, mercury pore size analyzer, and SEM were used, where yield, specific surface area and methylene blue number for adsorptive capacity, pore size, and appearance of the carbon were analyzed. Results show that gasification of unburned carbon produces larger adsorptive capability as well as a larger specific surface area. While reacting with CO2 at 1100 degrees C for 45 min, the methylene blue numbers reach to 54-154 mg/g, and the specific surface areas reach to 113-235 m2/g, which signify the actual utility. All carbons in gasification form new pores in a mutual size approximately 0.1-0.01 microm, whose majority affects the adsorptive capacity and specific surface area of the carbon. Besides, their group appearance complies with the obvious flow-like pattern, containing carbon ingredients with various degrees of crystallinity in the surface of unburned carbon. The new pore, conclusively, forms from the lower crystalline carbon ingredient that gasified with CO2 earlier, in the flow-like pattern. PMID- 15332676 TI - Effects of surface chemical and electrochemical factors on the dewatering characteristics of fine particle slurry. AB - Coal suspension was employed in vacuum filtration process to observe the dewatering characteristics of fine particle slurry. Generally, as the size of coal particle decreased, the specific resistance was increased and dewatering efficiency was reduced. Slurries containing the particles with a broad size range had a higher compressibility and could not filter easily if modified process was not applied. This study also investigated the effects of an anionic and a cationic surfactants on filterability by examining the surface property of coal particles. Dewatering was observed to be more efficient when cationic surfactant was used. To improve dewatering rate and to reduce the final moisture content of slurry, electrodewatering combined with vacuum filtration was applied. The effects of some different variables were examined, such as the type of surfactants, electrode (arrangements and materials), and the electrical parameters (voltage and current). By application of electrodewatering, dewatering efficiency was found to be enhanced compared to conventional vacuum filtration. The specific resistance was represented to be 0.3815 at 0 V, whereas it was shown to be 0.2113 and 0.1631 at 20 V under direct current and alternative current, respectively. PMID- 15332677 TI - Mixing characteristics and whey wastewater treatment of a novel moving anaerobic biofilm reactor. AB - A novel moving anaerobic biofilm reactor was used to treat whey wastewater. In this process, biofilm was grown on a plastic biofilm media module, which was vertically moved up and down in the bulk fluid. The objectives of the study were to investigate the mixing and performance characteristics of the new process in treating whey wastewater. The mixing efficiency was indicated by a dispersion number, D(L)/uL. D(L)/uL was up to 1.34, showing that the anaerobic reactor can be taken as a completely mixed reactor. At mesophilic conditions (35+/-2 degrees C), the admissible volumetric COD loading rate up to 11.6kg COD m(-3) day(-1) was achieved with the COD removal efficiency of 89% and the hydraulic retention time (HRT) of 1 day. When the HRT was 0.6 days, the volumetric COD loading rate was 15.2 kg COD m(-3) day(-1), but COD removal efficiency decreased to 81%. The percentage of methane (CH4) in the biogas was 63% on average and the yield of methane was 333.4 L CH4 kg(-1) COD removal at ambient conditions. PMID- 15332678 TI - Anaerobic fluidized bed reactor for the treatment of landfill leachates. AB - Treatability of the sanitary young landfill leachate in a pilot-scale anaerobic fluidized bed reactor during reactor startup and steady-state phases was investigated. All runs were carried out at 35 degrees C due to environmental conditions in the mesophilic anaerobic fluidized bed reactor (AFBR) while organic loading rate (OLR) was increased from 2.5 to 37gCOD/L-day during the 220 days of operation. The AFBR process attained steady-state conditions about on day 80 and a good and stable COD removal were achieved at about 90%. Biogas production in the bed continuously increased during the process. The mean specific biogas production was found 0.52 L biogas/gCODrem while the methane content was about 75%. The attached biomass concentration, measured as volatile solids rapidly increased as containing about 90% of the total biomass concentration. Furthermore, an increase in the suspended solid concentration was found as an evidence of biomass detachment from the media. PMID- 15332679 TI - Removal of metals from industrial sludge by extraction with different acids. AB - This study addresses the feasibility of removing Cu and Ni from sludges of printed circuit board (PCB) plants by acidic extraction. Citric acid, acetic acid, hydrochloric acid, nitric acid, and sulfuric acid at various concentrations were experimentally examined to determine preferred conditions for removing Cu and Ni. Experimental results indicate that the removal ratios of Cu were 57% (citric acid), 79% (acetic acid), 81% (hydrochloric acid), 91% (nitric acid), and 92% (sulfuric acid), and that of Ni was 74% (citric acid), 70% (acetic acid), 75% (hydrochloric acid), 92% (nitric acid), and 93% (sulfuric acid), using 1 N extractants. The removal ratios obtained using sulfuric acid or nitric acid at a particular concentration were in the order Pb> Ni> Cu> Zn. The kinetic extraction results revealed that the extraction of Cu and Ni by nitric acid and sulfuric acid exhibited a lagphase after 60min and the removal ratio was then around 90%. The kinetic results also revealed that the extraction rate constants of Cu and Ni increased with the concentration of the acid. The extraction rate constants of Cu and Ni followed the order sulfuric acid > nitric acid > acetic acid, and it also showed that Ni was more easily extracted than Cu from PCB sludge. The total percentages of Cu extracted by 1 N acetic acid, nitric acid, and sulfuric acid, determined in three replicate extractions were 78, 94, and 94%, and those of Ni were 89, 94, and 98%, respectively. The residue sludge, following three replicate extractions using 1 N acetic acid, nitric acid, and sulfuric acid, was further tested by toxicity characteristic leaching procedures (TCLP), and more than 15 mg/L of Cu was leached, exceeding the legal concentration in Taiwan. This observation reveals that the residue sludge following three replicate extractions by acids remained hazardous waste. Further treatment, such as solidification, must be performed before the sludge is disposed of. PMID- 15332680 TI - Mesoporous mineral columns for color removal from aqueous solutions. AB - The removal efficiency of natural minerals (e.g., zeolite and sepiolite) from aqueous solutions of azo dyes was investigated. The adsorption of three types of reactive dyes, Black, Red, and Yellow dissolved in distilled and/or deionized water on zeolite and sepiolite have been performed using column adsorption experiments. Distilled water containing 50 mg/L dyestuff was used to identify the ability of natural mesoporous minerals and their modified forms. The adsorption results indicate that both natural sepiolite and zeolite have limited adsorption capacities of the reactive dyes but are substantially improved upon modifying their surfaces with quaternary amines (HTAB). The removal of Yellow dye showed an excellent performance in sepiolite column compared to that in zeolite column. PMID- 15332681 TI - A two-stage SBR process for removal of organic substrate and nitrogen via nitrite type nitrification-denitrification. AB - A two-stage SBR process (TSSBR) was applied for the treatment of wastewater with high strength COD and nitrogen. Most of organic substrate was removed in the first stage SBR reactor (SBR1) under the aerobic condition. Subsequently the second stage SBR reactor (SBR2) firstly was operated under the aerobic condition for simultaneous nitrite-type nitrification and removal of a small amount of residual organic substrate. Nitrification was controlled to the nitrite-type nitrification. Then denitrification in which the wastewater was used as external carbon sources occurred in SBR2 under the anoxic condition. The experimental results indicated that two kinds of biomass with the different function were cultured in a TSSBR system, which was beneficial to increase the proportion of Nitrosomonas in the microorganism of SBR2. It avoided the inhibition of the high organic loading to nitrification and C/N was not the limiting factor affecting the nitrogen removal efficiency. The rate of COD degradation and nitrification was improved, 8 kg COD/kg MLSS/d and 0.25 kg NH4+-N/kg MLSS/d, respectively. The final effluent COD concentration further reduced because the organic substrate that was left in SBR1 effluent was removed in SBR2. Consequently, when compared to a single SBR process, a two-stage SBR (TSSBR) not only improved the treatment efficiency, but also saved the operational cost. PMID- 15332682 TI - Funding cuts could expose states to terrorist attacks. PMID- 15332683 TI - National medication safety managers see big picture. PMID- 15332684 TI - Pharmacists outside U.S. inch toward independent prescribing. PMID- 15332685 TI - Radio frequency tags for identifying legitimate drug products discussed by tech industry. PMID- 15332686 TI - New drugs and dosage forms. PMID- 15332687 TI - Computerized system for reminding prescribers to monitor warfarin therapy. PMID- 15332688 TI - Patient interest in receiving diabetes care in Hungarian community pharmacies. PMID- 15332689 TI - Compatibility of ketamine hydrochloride and fentanyl citrate in polypropylene syringes. PMID- 15332690 TI - Lack of evidence for ASHP's sterile-products guidelines. PMID- 15332691 TI - Opioids for painful diabetic neuropathy. PMID- 15332692 TI - Omalizumab: a recombinant humanized anti-IgE antibody for allergic asthma. AB - PURPOSE: The pharmacology, efficacy, dosage, adverse events, and economics of omalizumab are discussed. SUMMARY: Omalizumab, a recombinant DNA-derived humanized monoclonal antibody, binds to the C epsilon3 domain of immunoglobulin E (IgE) and forms complexes that inhibit the immune system's response to allergens by averting IgE-mediated inflammatory changes. Omalizumab exhibits a similar pharmacokinetic profile in adults, adolescents, and children. Omalizumab is indicated for adults and adolescents with moderate to severe persistent asthma who have a positive skin test or in vitro reactivity to a perennial aeroallergen and whose symptoms are inadequately controlled with inhaled corticosteroids. Because of the limited data regarding the safety and effectiveness of omalizumab in children, the drug is indicated for patients 12 years of age or older. The recommended starting dosage is 150-375 mg s.c. every two or four weeks. Dosages and frequency of dose administration are determined by total serum IgE level, measured before the start of treatment, and body weight. Omalizumab is generally well tolerated in adults and children with allergic asthma. Adverse events most commonly observed are injection-site reaction, viral infection, upper-respiratory tract infection, sinusitis, headache, and pharyngitis. Three large phase III clinical trials demonstrated that omalizumab is more effective than placebo in controlling moderate to severe allergic asthma in patients who have poor disease control or exacerbations despite recommended therapy. Currently, there are no clinical comparisons of omalizumab with other standard treatments for asthma; therefore, it is difficult to determine its overall place in therapy. CONCLUSION: Omalizumab should be considered as a second-line therapy for patients with moderate to severe persistent allergic asthma. PMID- 15332693 TI - Management of gestational diabetes mellitus and pharmacists' role in patient education. AB - PURPOSE: The pathophysiology, diagnosis, complications, and management of gestational diabetes mellitus (GDM) are discussed, along with considerations in setting up a pharmacist-run GDM education service. SUMMARY: GDM occurs when there is insufficient insulin secretion to counteract pregnancy-related decreases in insulin sensitivity. GDM can be diagnosed by using the same criteria used to diagnose types 1 and 2 diabetes mellitus (DM): a fasting blood glucose concentration of > 126 mg/dL on two separate occasions or a random blood glucose concentration of > 200 mg/dL on two separate occasions. Complications of GDM include maternal type 2 DM, maternal hypertension, macrosomia, shoulder dystocia, and neonatal hypoglycemia. GDM is managed with medical nutritional therapy (MNT), exercise, and therapy with human or synthetic insulin. The American Diabetes Association recommends starting insulin therapy when MNT fails to maintain plasma glucose concentrations at < or = 105 mg/dL during fasting, < or = 155 mg/dL one hour after eating, or < or = 130 mg/dL two hours after eating. A pharmacist interested in establishing a GDM education service must assess the feasibility of providing such education in his or her practice and whether such a program is needed. Other considerations are developing a curriculum, marketing the service, maintaining records, calculating costs, and obtaining reimbursement. CONCLUSION: GDM can have serious effects if not treated properly. A major part of managing GDM involves educating the patient about diet, exercise, blood glucose self monitoring, and insulin self-administration. A successful pharmacist-run GDM education service must have a market and prices sufficient to generate profit. PMID- 15332694 TI - Pharmacist surveillance of adverse drug events. AB - PURPOSE: The incidence of adverse drug events (ADEs), preventable ADEs, and potential ADEs was determined using pharmacist surveillance. Drug classes associated with ADEs were also identified. METHODS: The study was conducted in a 30-bed hospital ward of a Canadian teaching hospital between April 28, 2003, and May 26, 2003. All patients admitted to the general medicine service were eligible for study enrollment. A pharmacist performed surveillance to identify new or worsening symptoms, critical laboratory values, and medication errors. Surveillance consisted of daily communications with staff, daily chart reviews for all inpatients, and investigation of spontaneous incident reports. Data were collected to describe all identified outcomes. This information was rated independently by two clinicians to determine if the outcome was an ADE, a preventable ADE, or a potential ADE. Descriptive statistics were used to calculate outcome rates, which were reported as events per 100 patient-days. RESULTS: During 543 patient-days of observation, 24 ADEs occurred (4.4 per 100 patient-days), of which 14 were preventable (2.6 per 100 patient-days); 13 potential ADEs also occurred (2.4 per 100 patient-days). Of all ADEs, 3 (13%) were life threatening, 11 (46%) were serious, and 10 (42%) were significant. The 24 ADEs were associated with nine different drug classes. Four drug classes accounted for 17 ADEs (71%): antidiabetic agents, antibiotics, glucocorticoids, and sedatives and hypnotics. CONCLUSION: Pharmacist surveillance revealed that 4.4 ADEs occurred per 100 patient-days, over half of which were preventable. All preventable and potential ADEs occurred during the ordering and administration stages of medication delivery. PMID- 15332695 TI - Patterns of antihypertensive medication use in hemodialysis patients. AB - PURPOSE: Patterns of antihypertensive drug use in patients undergoing long-term hemodialysis therapy were studied. METHODS: Patients attending seven outpatient hemodialysis units in Ohio were eligible for the study if they had been receiving the treatment for at least three months. Demographic and clinical data were obtained from patient interviews and computerized databases, and blood pressure (BP) measurements were obtained before dialysis. Labeled names and dosages of antihypertensive drugs were recorded from containers the patients brought with them. Patients were asked to describe their adherence to the medications, their ability to afford the drugs, and their knowledge and beliefs about hypertension. Physical impairments in taking medication were also evaluated. RESULTS: The frequency of hypertension was 89% in the 270 participants. Antihypertensive drugs were prescribed for 76% of the patients; 25% required three or more drugs. Hypertensive patients who were not receiving antihypertensive drugs (14%) had significantly higher BP than patients who were. Calcium-channel blockers were prescribed for 60% of patients, angiotensin-converting-enzyme inhibitors for 33%, and beta-blockers for 34%. Eighty-three percent said the cost of drugs was never a problem, and 23% said they sometimes forgot to take their medication. Almost all patients said they would not stop taking antihypertensive drugs if they were feeling better and agreed that monitoring BP was important. Twenty-two percent could not read the warning on a standard tablet container, and 12% could not remove one tablet from the container. CONCLUSION: Multidrug antihypertensive therapy was common among hemodialysis patients and was associated with significantly lower BP; calcium-channel blockers were the most frequently prescribed agents. Most patients said they could afford drugs and reported good adherence to antihypertensive drug therapy. PMID- 15332696 TI - Prescribing practices for the treatment of hepatitis C in a Veterans Affairs medical center. PMID- 15332697 TI - Accuracy of splitting unscored valdecoxib tablets. PMID- 15332698 TI - Community pharmacists' responses to drug-drug interaction alerts. PMID- 15332699 TI - Pharmacist interventions in pain management. PMID- 15332701 TI - Stability of nimodipine solution in oral syringes. PMID- 15332700 TI - Increasing pneumococcal vaccination rates among adults with cystic fibrosis. PMID- 15332702 TI - Use of drug samples as a threat to seamless health care. PMID- 15332703 TI - Overview of geriatric pharmacy practice in a rehabilitation facility. PMID- 15332704 TI - Activity of mupirocin and 14 additional antibiotics against staphylococci isolated from Latin American hospitals: report from the SENTRY antimicrobial surveillance program. AB - A total of 1,346 Staphylococcus aureus (SA) and 498 coagulase-negative staphylococcal (CoNS) strains isolated from 11 Latin American medical centers between 2000 and 2001 were tested against mupirocin and other antimicrobial agents by reference broth microdilution method as part of the SENTRY Antimicrobial Surveillance Program. Oxacillin resistance (OR) was detected in 38.6% of S. aureus and in 78.1% of CoNS. The overall resistance rate to mupirocin was low among S. aureus (3.1%; MIC > or =8 microd/ml) but significantly higher among ORSA compared to oxacillin-susceptible SA (5.4% versus 1.7%; p <0.001). Mupirocin-resistant S. aureus strains were detected in 9 of 11 centers, with individual center rates varying between 1.8 and 15.7%. Mupirocin resistance rates were high among CoNS (27.5%) and varied widely (10.0 to 48.9%) among the monitored Latin American medical centers. Mupirocin resistance rates appear to be increasing and routine monitoring for potential resistance seems prudent. PMID- 15332705 TI - Reassessment of cefaclor breakpoints for Haemophilus influenzae. AB - We previously reported that standard methods overestimate cefaclor minimum inhibitory concentrations (MICs) for Streptococcus pneumoniae due to in vitro chemical instability. This study aimed to ascertain if standard methods accurately measure cefaclor MICs to Haemophilus influenzae. Cefuroxime was used as a comparator. Standard NCCLS broth microdilution and E-Test MICs were determined for eight isolates of H. influenzae. Kill curves determined the "bacteriostatic" MIC, defined as the concentration showing no significant growth or kill over six hours taking into account cefaclor instability. On average, cefaclor and cefuroxime bacteriostatic MICs were 0.2 x MIC and 0.6 x MIC determined by NCCLS methodology respectively. The mean MIC determined by NCCLS methodology was 3.0 mg/L for cefaclor and 0.8 mg/L for cefuroxime. Cefaclor MICs by NCCLS methodology were overestimated due to chemical instability over 18-24 hours. The bacteriostatic MICs by kill curve were not significantly different from those of cefuroxime. PMID- 15332706 TI - Evaluation of moxifloxacin activity in vitro against Mycobacterium tuberculosis, including resistant and multidrug-resistant strains. AB - The new quinolone moxifloxacin was tested against 86 strains of Mycobacterium tuberculosis including 13 resistant and 4 multiresistant strains. The antimicrobial susceptibility was tested, in parallel, using two different liquid media, the radiometric Bactec 12B and the Mycobacteria Growth Indicator Tube (Becton Dickinson, USA). All strains but two were susceptible at 0.5 microg/ml of moxifloxacin; for the remaining two strains, both multidrugresistant, the minimal inhibitory concentrations (MIC) were =2 and >4 microg/ml respectively. Our data confirm the high antitubercular in vitro activity of moxifloxacin. PMID- 15332707 TI - Molecular characterization of beta-lactam resistance of Salmonella isolates from pediatric patients in Romania. AB - The molecular characterization of 16 clinical isolates of Salmonella enterica (14 serotype Typhimurium and 2 serotype Kingston) obtained between January and June 1999 from feces of children hospitalized in Iasi, Romania were genotypically compared by pulse field gel electrophoresis of XbaI restricted bacterial DNA. The majority of the clinical isolates (12/16) belonged to cluster A and (4/16) to unrelated strains, correlating to the OMP profile. Two major different patterns of beta-lactamases were identified: the first with pI of 5.4, 8.2 in 6/16 strains and the second with pI of 5.4 in 5/16. The blaTEM beta-lactamase was identified in 14/16 of the clinical isolates and the blaSHV-5 gene in one strain. We concluded that extended spectrum beta-lactamase (ESBL) with pIs of 8.2 was the most frequent enzyme produced by serotype Typhimurium isolates which were related. PMID- 15332708 TI - PER-1 production in a urinary isolate of Providencia rettgeri. AB - A Providencia rettgeri strain resistant to extended-spectrum cephalosporins and intermediate to aztreonam was isolated from the urine of a patient hospitalized in the urology clinic of SSK Educational Hospital in Ankara. Clavulanic acid restored the activity of extended-spectrum cephalosporins, suggesting that the strain was harboring an extended-spectrum beta-lactamase. Since the PER-1 enzyme is widespread in Turkey, and had been already detected in a related species such as Proteus mirabilis, the Providencia strain was suspected of harboring a PER-1 enzyme, which was indeed detected by PCR. This is the first description in a P. rettgeri isolate of a PER-1 enzyme which is widespread among Acinetobacter baumanni and Pseudomonas aeruginosa strains in Turkey. PMID- 15332709 TI - The contribution of a novel ribosomal S12 mutation to aminoglycoside resistance of Escherichia coli mutants. AB - This study characterised the contribution of a novel ribosomal S12 mutation to aminoglycoside resistance in Escherichia coli via step-wise mutation analysis. Mutants of E. coli NCTC 10418 were selected in four separate progressive series (I-IV) on plates containing increasing aminoglycoside (streptomycin, neomycin, gentamicin, tobramycin, and kanamycin) concentrations. Minimum inhibitory concentrations (MICs) of these aminoglycosides were established for the most resistant mutants in each series. There was no cross-resistance between streptomycin and the other aminoglycosides tested; however there was cross resistance between the neomycin, tobramycin and kanamycin resistant mutants. DNA sequencing of a 423bp region of the rpsL gene encoding S12 revealed a novel Lys87 ->Glu mutation in the streptomycin selected resistant mutants, while there were no S12 mutations in resistant mutants resulting from selection with neomycin, gentamicin, and tobramycin and kanamycin. PMID- 15332710 TI - Antiplasmodial activity of acetogenins and inhibitory effect on Plasmodium falciparum adenylate translocase. AB - Three Annonaceous acetogenins exhibited in vitro antimalarial activities on a chloroquine-resistant Plasmodium falciparum strain, with IC50s ranging from 5 to 10 microM. Structure-activity relationships showed that maximal antimalarial activity occurred in the presence of at least one tetrahydrofuran moiety and a synergistic action with chloroquine was observed. These acetogenins partially inhibited the P. falciparum adenylate translocase. PMID- 15332711 TI - Immunomodulating activity of rifampicin. AB - It has been shown that some antibiotics can modify cytokine production. We have examined the effect of rifampicin on secretion of interleukin-1beta (IL-1beta), IL-6, IL-10, and tumor necrosis factor alpha (TNF-alpha) by lipopolysaccharide (LPS)-stimulated or heat killed staphylococci (Pansorbin) stimulated monocytes. Secretion of IL-1beta and TNF-a were significantly inhibited (P<0.002) whereas secretion of IL-6 and IL-10 were significantly increased (P<0.003) by rifampicin treated mononuclear cells. Rifampicin had immunomodulatory effects through its capacity to alter the secretion of tested cytokines by human monocytes. PMID- 15332712 TI - Meropenem monotherapy for the treatment of hospital-acquired pneumonia: results of a multicenter trial. AB - The efficacy and tolerability of meropenem as empirical treatment in patients with hospital-acquired pneumonia was determined in a prospective, open-label, non randomized trial. Patients from 28 centers in the USA received meropenem 1 g every 8 h intravenously. Of 255 patients enrolled, 111 were evaluable for efficacy, including 60 patients with ventilator-associated pneumonia. At end of treatment 74% of patients had a satisfactory clinical response and 64% had this response at follow-up, which could last up to 28 days after treatment. In patients with ventilator-associated pneumonia, a satisfactory clinical response was observed in 68% at the end of treatment and 63% at follow-up. The overall satisfactory response rate for individual pretreatment pathogens ranged from 65% to 100%. This study demonstrates that meropenem monotherapy is effective and well tolerated for patients with hospital-acquired pneumonia, including a subgroup of patients with ventilator-associated pneumonia. PMID- 15332713 TI - Effects of paclitaxel and docetaxel on EGFR-expressing human carcinoma cells under normoxic versus hypoxic conditions in vitro. AB - Human malignant tumors, such as non-small lung, breast, ovarian, head and neck, prostate, stomach and colorectal cancers express a number of growth factor receptors (e.g. EGFR or EGFR family members) that are regulated by tumor hypoxia and contribute to tumor growth and failure of cytotoxic therapy. Paclitaxel and docetaxel are indispensable substances in the treatment of these tumors. Despite the active clinical use of taxanes, little is known about their cytotoxic activity under hypoxia. The aim of the present work was to compare the cytotoxic effect of taxanes, paclitaxel and docetaxel on the EGFR-expressing carcinoma cell lines A431, MDA-MB-231 and NCI-H358 under normoxic and hypoxic conditions. The two taxanes caused different cell cycle distribution and varying aneuploid cell formation under hypoxia. EGFR-overexpressing carcinoma cells showed hypoxia to severely affect the cytotoxicity of paclitaxel, whereas docetaxel preserved its tumor cell-killing activity even at lowest concentrations (0.5 nM), as was observed for both taxanes under normoxia. PMID- 15332714 TI - Caffeic acid phenethyl ester ameliorated ototoxicity induced by cisplatin in rats. AB - Caffeic acid phenethyl ester (CAPE), an active component of propolis, exhibits antioxidant properties. This experimental study was designed to determine the effect of CAPE on ototoxicity induced with cisplatin. Twenty-four adult Wistar albino rats were divided into four groups: cisplatin (n=6), saline (n=6), CAPE (n=6), and cisplatin plus CAPE (n=6). Rats were tested before and 5 days after cisplatin treatment with or without chemo protection. The Distortion Product Otoacoustic Emissions (DPOAEs) were elicited from the control and experimental animals utilizing the standard commercial Otoacoustic Emission (OAEs) apparatus. The animals in all groups were sacrificed under general anesthesia on the fifth day following last OAE measurements. For biochemical investigations, the blood samples were drawn from inferior vena cava. On day 0, the initial baseline DPOAEs measurement results presented similar values while comparing the groups in drug free phase (p>0.05). On day 5, intrasubject measurement parameters of DPgrams and I/O functions of cisplatin group were significantly deteriorated (p<0.05). The second measurements of the other groups revealed no significant differences between their DPgrams and I/O functions in all frequencies (p>0.05). Among the biochemical parameters, plasma xanthine oxidase (XO) activity was found to be more elevated in the cisplatin group than the saline group (p<0.05). CAPE led to more decreased XO activity than cisplatin (p<0.05). The results of this study show that prophylactic administration of CAPE for cisplatin ototoxicity ameliorated hearing deterioration in rats. PMID- 15332715 TI - Role of low-dose 2-CdA in refractory or resistant lymphoplasmocytic lymphoma. AB - Cladribrine (2-CdA), a purine analogue active on both dividing and resting lymphocytes, plays an important role in the treatment of indolent lymphoproliferative malignancies such as Hairy Cell Leukemia (HCL), Chronic Lymphocytic Leukemia (CLL), Lymphoplasmocytic Lymphoma (LPL), Waldenstrom's Macroglobulinemia (WM). With the aim of evaluating the efficacy and toxicity of low dose 2-CdA, 15 lymphoplasmocytic lymphoma patients, not eligible for more aggressive or standard therapies, because of age or poor performance status, were treated with the drug at a dose of 5 mg/m2, once a week for six total courses. All patients showed disease progression. Fourteen patients were valuable for response. In eleven out of these 14 (85.7%) disease progression stopped, with 21% having good hematological responses (one CR and two PR). The treatment was generally well tolerated, without serious infectious events. This schedule may be appropriate for the management of patients where the aim of the treatment is control of disease progression. PMID- 15332716 TI - Weekly vinorelbine and docetaxel as second-line chemotherapy for pretreated non small cell lung cancer patients: a phase I-II trial. AB - Docetaxel was proven to be effective as second-line therapy for patients with advanced NSCLC after failure of platinum-based front-line chemotherapy. We designed this phase I/II study to define the Maximum Tolerated Dose of weekly docetaxel combined with weekly vinorelbine, and subsequently evaluate tolerability and activity of this schedule in NSCLC patients who were progressive after treatment with either cisplatin and gemcitabine or carboplatin and paclitaxel regimens. To be eligible for the study, patients were required to have a WHO performance status < or =2, failure after at least two cycles of first platinum-based chemotherapy, and no prior treatment with docetaxel and vinorelbine. A total of 27 patients were enrolled in this phase I/II study. A weekly docetaxel dose of 25 mg/m2 was recommended in combination with fixed vinorelbine dose of 20 mg/m2, and 24 patients were treated at this dose level. Severe neutropenia (62%) and febrile neutropenia (29%) were the most frequent toxicities, with 83% of patients requiring dose modification or delay. In the phase II study, 5 (21%) patients obtained a partial response, 8 (33%) patients had stable disease, whereas 10 (42%) patients progressed. After a median follow up of 18.7 months, median survival was 8 months, with 30% surviving at 1 year. Regardless of the use of weekly docetaxel schedule, this regimen was highly myelosuppressive, and did not seem to improve response rate and survival compared to single-agent docetaxel. No further developments of this schedule are warranted. PMID- 15332717 TI - Efficacy of the combination ampicillin plus ceftriaxone in the treatment of a case of enterococcal endocarditis due to Enterococcus faecalis highly resistant to gentamicin: efficacy of the "ex vivo" synergism method. AB - The case of a patient with highly gentamicin-resistant Enterococcus faecalis endocarditis treated with an ampicillin + ceftriaxone combination is described. We have designed a method to evaluate synergism between the antibacterial activity of patient's serum taken during a given antibiotic regimen (ampicillin) to which another antibiotic (ceftriaxone) is added in vitro. In this patient the two-drug combination was able to stop the bacteremia and prevent the infection of the prosthetic valve. PMID- 15332718 TI - Melioidosis in a traveller from Thailand: case report. AB - A 42-year old Italian male with type 2 diabetes and HCV-related chronic hepatitis spent 6 months in Thailand. After his return in June 2002 he was admitted to the Infectious Diseases Unit of the Hospital of Livorno (Italy) because of fever, chest pain and skin abscesses in the legs. Chest X-rays and CT scan revealed multiple bilateral cavitary lesions in the lungs. Ultrasonography and CT scan showed numerous subcentimetric spleen abscesses. Burkholderia pseudomallei was isolated from the cutaneous lesions and sputum and thus melioidosis was diagnosed. A 6-week course of i.v. ceftazidime plus oral doxycycline was given during the acute phase of the illness. The in vitro susceptibility testing showed that long-term (20 weeks) antimicrobial therapy with doxycycline and moxifloxacin was required. Complete resolution of pulmonary and spleen lesions was obtained within 6 weeks of therapy and of cutaneous abscesses in 10 weeks. No significant side effects were noted during the follow-up period using this scheme of antimicrobial therapy. PMID- 15332719 TI - Azithromycin: 4.5- or 6.0-gram dose in the treatment of patients with chronic prostatitis caused by Chlamydia trachomatis--a randomized study. PMID- 15332720 TI - P16INK4a overexpression is associated with poor clinical outcome in ovarian carcinoma. PMID- 15332721 TI - In search of the good life: a cultural odyssey in the East and West. AB - In the first part of this article, the authors argue that any conception of well being is culturally embedded and depends on how the notions of "well" and "being" are defined and practiced in different cultural communities. To support this argument, they conduct a comprehensive review of the empirical literature, which shows that members of individualistic and collectivistic cultures differ in their emotional experience of well-being. This difference has been traced to the indigenous psychology of selfhood in those cultures. In the second part of the article, the authors debunk the myth of infinite cultural variability, while retaining the basic insight concerning the cultural constitution of well-being. They develop a theoretical model of well-being, in which agency and communion are universal dimensions of well-being, and people in different cultures require different combinations of agency and communion to experience well-being. Following this theoretical development, the authors examine cultural impediments to the good life in individualistic and collectivistic cultures. The mutual incompatibility of agency and communion is ruled out as a possible cause of why it is so difficult to live the good life. Instead, the authors propose a dialectical synthesis of agency and communion as a way of dealing with this challenge of living the good life. PMID- 15332722 TI - Individuation and relatedness: not opposing but distinct and complementary. AB - In this article, the author aims to contribute to a better understanding of the association between relational and individuational self-orientations and the roles they play in the self-system. The author highlights the controversial assumptions regarding the opposite or distinct nature of the orientations' association and explores how they relate to each other and to some self- and family-related variables by a questionnaire study. On the basis of the Balanced Integration-Differentiation model (E. O. Imamoglu, 1998), relatedness and individuation were hypothesized to refer to distinct and complementary self orientations; the former was expected to be associated more with affect-related variables (i.e., perceived parental love-acceptance, self- and family satisfaction), whereas the latter was expected to be associated more with intrinsic-motivational variables (i.e., need for cognition and negatively with perceived parental control). University students (N = 274) from Turkey participated in the study. Results indicated that (a) individuation and relatedness were not negatively correlated; (b) perceived parental love acceptance predicted relatedness both directly and indirectly through the mediation of self- and family satisfaction, whereas perceived parental control predicted (negatively) individuation indirectly through the mediation of need for cognition, a strong predictor of individuation; and (c) being both related and individuated appeared to be associated with optimal psychological functioning, the implications of which are discussed. PMID- 15332723 TI - [Experience in a suburban hospital with premature termination of pregnancy in the second trimester]. AB - The present study was performed between 1992 and 1998 and had the aim of examining the activity and side-effects of intravenously administered sulprostone for the induction of abortion in the second trimester. 160 patients were admitted to hospital. 3 were submitted to a hysterectomy; 3 underwent an abortion after cervical priming with Prostin E2 Gel (0.25 mg/ml), 154 were repeatedly treated with an infusion of sulprostone for 6 hours (500 microg Nalodor, 3 A/1000 ml 0.9% NaCl, 250 microg/h (equivalent to 4.17 microg or ca. 55 drops per min.). The mean interval for induction of abortion was 16 h 56 min; 53% of the abortions occurred within 12h; 95% of the abortions occurred within 48 h. Two inductions were unsuccessful. 12 women suffered from atonic bleeding. This study demonstrated that sulprostone infusions are suitable for the induction of abortions in the second trimester and that side-effects are rare. Particular attention must be paid to the complication of bleeding. PMID- 15332724 TI - [Does red wine protect from myocardial infarction]. PMID- 15332726 TI - [Odynophagia after viral infection]. PMID- 15332725 TI - [Emergency landing due to a passanger with chest pain, dyspnea, and vomiting]. AB - A 20-year old student had suffered since 3 years from diabetes mellitus type I, which was well-controlled by insulin-pump therapy. During a flight from Moscow to Los Angeles, the student all of a sudden had chest pain, dyspnea, and he vomitted repetitively--emergency landing at Zurich airport was necessary. The student presented at the emergency unit in a poor general condition with tachypnea (32/min) and tachycardia (136/min). Arterial blood gas analysis showed severe metabolic acidosis (pH 7.04), while pulmonary or cardiac disease could be ruled out. Diabetic ketoacidosis was caused by the pump running short of insulin. Treatment included rehydration and administration of insulin. Administration of insulin by an insulin-pump allows to continuously and flexibly adjust the dosage according to the requirement of the body. Interruption of insulin administration can cause, however, relatively fast ketoacidosis because exclusively short-acting insulin is used. PMID- 15332727 TI - [Painful exanthema with fever. 37-year-old patient. Idiopathic Sweet syndrome (acute febrile neutrophilic dermatosis)]. PMID- 15332728 TI - [A tropical plant for common cold symptoms]. PMID- 15332729 TI - College students' knowledge, beliefs and attitudes towards AIDS in predicting their safe sex behaviour. AB - We tested the applicability of the health belief model (HBM) in predicting college students' intentions to use condoms and condom use and to determine whether or not college students' attitudes toward AIDS would influence their decision to use condoms. United States university students (282) responded to a closed-format questionnaire about condom use, following an elicitation open-ended survey. Multiple regression analysis was performed to determine the predictive power of the HBM components. Of HBM components, cue to action, barrier and benefit were the most powerful predictors of both condom use intentions and behaviour. Students' beliefs and attitudes toward AIDS did not influence their decision to use condoms. PMID- 15332730 TI - Bioenvironmental and meteorological factors related to the persistence of malaria in Fayoum Governorate: a retrospective study. AB - The present study was conducted to determine the magnitude of the malaria problem in Fayoum Governorate and to ascertain the determinant factors behind its persistence in this area. Malaria records in Fayoum Governorate for the period 1971-1999 were reviewed retrospectively. The data show that no Plasmodium vivax cases were recorded during 1997-1999 and prediction up till 2004 indicated no positive cases expected. No P. falciparum cases were detected in 1998 and 1999. The prediction trend for this species showed an apparent but not statistically significant decrease in the incidence. The factors responsible for the persistence of malaria in this governorate are discussed. PMID- 15332731 TI - Diagnosis of cystic echinococcosis: ultrasound imaging or countercurrent immunoelectrophoresis? AB - Cystic echinococcosis is a major zoonotic diseases in the Islamic Republic of Iran. This study was carried out in 3 general hospitals in Shiraz. We examined the records of all 1227 surgical patients with a surgically-proven diagnosis of cystic echinococcosis for the 20-year period 1978-98. The results of countercurrent immunoelectrophoresis were compared with pathology and ultrasound reports to determine whether serological tests could be helpful for diagnosis. Countercurrent immunoelectrophoresis could detect only 62.0% of cases, whereas the pathology and ultrasound results were positive for 96.3% of cases. This study confirms the usefulness of ultrasound and suggests that only in doubtful cases would countercurrent immunoelectrophoresis be useful for diagnosing cystic echinococcosis. PMID- 15332732 TI - Seroepidemiological study of visceral leishmaniasis among humans and animal reservoirs in Bushehr province, Islamic Republic of Iran. AB - Using direct agglutination tests, a survey of visceral leishmaniasis was carried out among children and adults from 13 villages and from nomadic tribes in Bushehr province during 1998-99. Of the 1496 plasma samples, the overall seropositive rate (titres > or = 1:3200) was 3.4%. Almost all cases (94.1%) were in children under 10 years old. Eighteen patients were diagnosed with kala azar; fever and splenomegaly were the predominant signs and symptoms. Parasitology and serology examinations of local animals identified dogs and jackals infected with Leishmania infantum. Suggestions for control of visceral leishmaniasis in this area are to eliminate stray dogs, identify cases among humans and suspected leashed dogs, and treat infected individuals. PMID- 15332733 TI - Risk factors for vaginal trichomoniasis among women in Basra, Iraq. AB - We investigated Trichomonas vaginalis infection among 352 women with vaginal discharge, 46 were found to be infected, an infection rate of 13%. There were no significant differences in the isolation rate of T. vaginalis in women according to occupation, educational level, economic status, age, marital status, parity, menstrual status and contraception use. The difference in the isolation rates of T. vaginalis in women with a history of abortion (7.6%) and in women with no history of abortion (15.7%) was statistically significant. PMID- 15332734 TI - Gram stain versus culture in the diagnosis of vulvovaginal candidiasis. AB - We evaluated Gram stain as a rapid diagnostic method for vulvovaginal candidiasis (VVC). Vaginal swabs were taken from 100 pregnant women and subjected to Gram stain and culture; 39% were diagnosed as having VVC (29% had symptomatic VVC and 10% had asymptomatic VVC). Candida albicans was isolated from 94.9% of all cultures while other Candida species were isolated from 5.1% of cases. Multigravidae were significantly more affected than primigravidae. Gram stain is a valuable method in rapid accurate diagnosis of symptomatic VVC and even superior to culture as it demonstrates the invasive forms of the yeast. However, its low sensitivity could restrict its use in routine practice. A combination of culture and Gram stain is the ideal approach for the diagnosis of VVC. PMID- 15332735 TI - Reproductive health of married adolescent women in squatter areas in Alexandria, Egypt. AB - Through a house-to-house survey, 455 married adolescent women (< 20 years) and 455 married women aged 20-45 years from squatter areas in Alexandria were interviewed. Determinants of early marriage were consanguinity, illiteracy and unawareness of reproductive health. Of the currently pregnant adolescents, 22% were receiving antenatal care compared with 40% of the older women. Family planning methods were currently being used by 39% of the adolescents and 63% of the older wives. Determinants of non-use were: adolescence, illiteracy and previous miscarriage/stillbirth. There is a need to increase public awareness of reproductive health and change the behaviour of both groups of women in this under-privileged sector. PMID- 15332736 TI - [Contraceptive use prevalence in a suburban zone around Sousse (Tunisia)]. AB - A contraceptive prevalence survey was conducted among 1277 women of reproductive age, to explore knowledge of and attitudes to contraceptive use and study factors influencing contraceptive use. We found that family planning methods were generally well known. The prevalence of modern contraceptive use was 53.6%, IUDs being the most commonly used followed by sterilization and pills. The public sector was the main source for obtaining contraceptives although pills and condoms tended to be obtained from the private sector. There was a significant correlation between contraceptive use and urban residence. PMID- 15332737 TI - Infant mortality in Arab countries: sociodemographic, perinatal and economic factors. AB - The infant mortality rates for 1978 and 1998 of 16 Arab countries in the Eastern Mediterranean region were studied. The data were extracted from World Health Organization and United Nations Children's Fund sources. The impact of demographic, social, perinatal care and economic indicators on infant mortality rates in 1998 was studied using Spearman rank coefficient to detect significant correlations. All countries, except Iraq, showed a sharp decline in rates from 1978 to 1998. Infant mortality rates were directly related to population size, annual total births, low birth weight and maternal mortality ratios. Also, infant mortality rates were inversely related to literacy status of both sexes, annual gross national product per capita and access to safe drinking-water and adequate sanitation facilities. PMID- 15332738 TI - Incidence and determinants of low birth weight in Syrian government hospitals. AB - The incidence and determinants of low birth weight were studied in a stratified sample of 10,585 deliveries at 18 Syrian government hospitals. For each low-birth weight baby born, two normal-birth-weight babies were recruited as a comparison group. A questionnaire was given to mothers and clinical examinations were performed on the neonates. The incidence of low birth weight was 6.6%. It varied according to geographic region and hospital. The study confirmed that low birth weight could be the result of prematurity. Factors amenable to intervention, such as birth interval, maternal nutrition, maternal weight and smoking, appeared to have a role in low birth weight. PMID- 15332739 TI - Disease severity associated with cystic fibrosis mutations deltaF508 and S549R(T- >G). AB - We compared the clinical severity associated with the two cystic fibrosis (CF) mutations S549R(T-->G) and deltaF508. Clinical and biochemical variables of CF were compared in two age- and sex-matched groups of CF children in the United Arab Emirates (UAE). The clinical severity of mutations S549R(T-->G) and deltaF508 showed comparable patterns, with very low Shwachman scores and high sweat chloride levels. We conclude that patients homozygous for the CF mutations deltaF508 and S549R(T-->G) have a severe clinical presentation and illness and are indistinguishable on clinical grounds. PMID- 15332740 TI - Cardiac and ocular manifestations in Egyptian patients with mucopolysaccharidoses. AB - Cardiac and ocular manifestations were evaluated in 21 patients clinically suspected of mucopolysaccharidosis. After electrophoresis analysis of urinary glycoaminoglycans, 3 patients were excluded because their results did not correlate with any known type of mucopolysaccharidosis. Echocardiography revealed abnormal findings in 11 patients (61.1%). The mitral valve was the most commonly affected valve; 7 patients (38.9%) had thickened mitral valve and 6 had mitral regurge. Corneal opacities were found in 3 patients (16.7%) and progressive increase in intraocular pressure in 1 patient (5.6%), while fundus examination showed early optic atrophy in 1 patient (5.6%) and bilateral papilloedema in 2 patients (11.1%). PMID- 15332741 TI - Cartilage proteoglycan aggrecan as a predictor of joint damage in juvenile rheumatoid arthritis. AB - Aggrecan was measured in the sera of 31 children with juvenile rheumatoid arthritis and in the synovial fluid of 10 of them. Patients were evaluated at baseline and 3 months later. Radiographs were repeated also after 1 year. As comparison, 15 apparently healthy children with no disease and 10 children with arthritis due to other collagen vascular diseases were studied. Baseline serum aggrecan was significantly higher in juvenile rheumatoid arthritis patients compared to controls and other patients. On re-evaluation, a significant drop in serum aggrecan from baseline values coincided with a significant drop in clinical and laboratory indices of active inflammation. Serum aggrecan can help to assess the extent of cartilage destruction and is useful as a prognostic tool to predict joint damage in patients with juvenile rheumatoid arthritis. PMID- 15332742 TI - A community-based study of common hereditary blood disorders in Oman. AB - We assessed the prevalence of three common hereditary blood disorders (sickle cell and beta-thalassaemia traits and glucose 6-phosphate dehydrogenase deficiency) among the Omani population. We interviewed a representative sample of 6103 Omani households and blood samples from 6342 children aged 0-5 years were collected. About 27% of Omani males had inherited glucose-6-phosphate dehydrogenase deficiency (compared with 11% of females) while countrywide prevalence rates for the sickle-cell and beta-thalassaemia traits were estimated to be 5.8% and 2.2% respectively and showed no significant gender differences. There was a significant association between all three disorders and region of the country. PMID- 15332743 TI - Breast cancer in south-east Republic of Yemen. AB - A retrospective study of breast cancer was carried out using the treatment registry of Aden Health Office and archives of Al-Gamhoria Teaching Hospital from January 1989 to December 1996 and the records of patients registered at Aden Cancer Centre from January 1997 to December 1998. The data analysed included age, sex, residence, breast affected and type of cancer. There were 227 patients with breast cancer registered between January 1989 and December 1998; 225 were female and 2 were male. Most of the patients (81.5%) had infiltrating ductal carcinoma. About 90% of the patients had lymph node involvement. The age groups most affected were 30-39 years and 40-49 years. PMID- 15332744 TI - Medical waste and health workers in Gaza governorates. AB - The study was conducted among health workers in Gaza to identify and highlight the problem of medical waste management. Data were collected through a questionnaire (given to 400 health workers), a checklist and interviews (with 16 decision-makers). Results show that there is no system for medical waste management in Gaza. Segregation is done only for sharps and there are no colour coded bags. Medical waste is stored and disposed of with domestic waste in primary health care clinics and is incinerated in hospitals, but there are no emission control or safety measures. There are some gaps in knowledge of health care workers, and current practices are inadequate. However there is generally a positive attitude to improving medical waste management among those surveyed and interviewed. A national programme for medical waste management is essential in Palestine. PMID- 15332745 TI - Joining the nursing profession in Qatar: motives and perceptions. AB - We aimed to identify why female students in Qatar decide to become nurses and how the students perceived the community attitude towards nursing. A self administered anonymous questionnaire was distributed to all (57) female students of the four academic classes of the Nursing Unit, University of Qatar for the academic year 1999-2000. The two commonest reasons for joining the nursing profession were an interest in medical services and the humanitarian nature of nursing. There were 33 (57.89%) students who considered there was a negative community attitude towards nursing mainly due to the presence of male patients and colleagues and the working hours. A mass media campaign and govemmental support were two strategies suggested to change this. PMID- 15332746 TI - Epidemiology of hepatitis B virus infection in the Middle East. AB - In this article, the epidemiology of hepatitis B (HBV) infection in the Middle East is reviewed. The prevalence of HBV carrier status in the region is considered and the modes of transmission of infection discussed. The risk factors associated with HBV infection are examined and preventive measures against HBV infection in countries of the region are presented. The review is based on information from two sources--publications in the international literature on the epidemiology of HBV infection in Middle East countries, identified by searching MEDLINE and POPLINE, and Middle East country and regional reports on viral hepatitis. It is evident that HBV infection is a serious public health problem in the region and many questions specific to the region remain unanswered. Community based surveys are recommended as they provide more accurate information that can be generalized than hospital-based surveys. PMID- 15332747 TI - The antenatal care debate. AB - The antenatal care debate has focused on the frequency of antenatal care, its content, continuity, quality, organization, effectiveness and impact on morbidity and mortality. Established antenatal care schedules have been called into question. Randomized controlled trials comparing reduced schedules with routine antenatal care have shown similar or better outcomes for the reduced protocols. Furthermore, midwives, with or without physicians, can provide continuous maternity care comparable to obstetrician-led care. Some women disliked the new protocol, but a reduced protocol of high quality provided by competent midwives is cost-effective, spares scarce physicians and ensures women's satisfaction. It is recommended globally. PMID- 15332748 TI - Overview of the water and wastewater reuse crisis in the Eastern Mediterranean Region. AB - Many countries of the Eastern Mediterranean region have very limited water resources. The situation will worsen in the future as populations grow and demands rise, and water/wastewater reuse will become imperative. This paper reviews the current situation in the Region as regards water use/reuse and management, and gives suggestions for how to ensure sufficient water resources for the future PMID- 15332749 TI - Post kala azar dermal leishmaniasis in Sudan. AB - Post kala azar dermal leishmaniasis (PKDL) is a condition that develops after treatment of kala azar. We report on 42 patients with suspected PKDL, 40% of whom were children. Diagnosis was made though investigation of family history of kala azar, clinical examination and the use of laboratory investigations, such as skin smear, skin biopsy, bone marrow aspiration and the leishmanin skin test. Regarding the lesions, 24 patients (57%) had papular lesions, 10 (24%) had hypopigmented maculopapular lesions and 8 (19%) had nodular lesions. The lesions of PKDL may be confused with other dermatological diseases and therefore it is important that clinicians and pathologists collaborate in diagnosing such cases. PMID- 15332750 TI - Haemolytic disease of the newborn caused by rhesus isoimmunization (anti-c). PMID- 15332751 TI - Perifollicular purpura must not be forgotten in scurvy: case reports. PMID- 15332752 TI - Parents in the anesthetic induction room: the preferences of parents in Kuwait. AB - OBJECTIVE: To ascertain whether the parents in Kuwait desire to be present during the induction of anesthesia in their children and to obtain the parents' views on factors associated with such presence. METHODS: The parents of 80 children scheduled for surgery under general anesthesia were given asked to complete a questionnaire. RESULTS: An overwhelming 88.75% of parents wished to he present in the anesthetic room during induction. CONCLUSIONS: Parents in Kuwait should he allowed to be present during the induction of anesthesia in their child. Protocols should be drafted accordingly and must include other child-friendly measures to reduce anxiety among parents and children in Kuwait. PMID- 15332753 TI - Reasons for underreporting of notifiable diseases by Syrian paediatricians. AB - We investigated reasons for underreporting of notifiable diseases among Syrian paediatricians. Self-administered questionnaires and self-addressed envelopes were sent to all paediatricians listed in the recent records of the Syrian Medical Association. Of 723 questionnaires, 55 were returned by the post office due to an incorrect address. Only 160 questionnaires (23.9%) were answered. Approximately 50% of paediatricians gave two reasons for underreporting: lack of reporting forms and ignorance of reporting telephone numbers. Nearly 70% indicated that the most important improvements would be the availability of easy reporting forms and a condensed and feasible list of notifiable diseases. PMID- 15332754 TI - Haemophilus influenzae type b diseases in children: a pre-vaccination study. AB - Haemophilus influenzae type b (Hib) can now be prevented by vaccination. We present the clinical and laboratory characteristics of acute invasive H. influenzae diseases in children admitted over a 4-year period to a tertiary paediatric ward of the Al-Ain medical district hospital, before vaccination became available in the United Arab Emirates. In all, 38 children had bacteriologically proven H. influenzae invasive diseases and all the isolates were serotype b. Meningitis was diagnosed in 60.5% of the children and 66% of the studied children were under 12 months. There were no deaths but substantial morbidity occurred in 12 children. PMID- 15332755 TI - Varicella susceptibility among children and healthy adults in the United Arab Emirates. AB - The serological evidence of varicella zoster virus infection was determined among healthy individuals from infancy to 47 years of age living in this region. Of 648 people, 126 (19.4%) had no detectable antibody and were susceptible to infection. The overall adult seroprevalence rate was 81.3%. The rate among Emirati citizens increased with age; < 10 years, 45.8%; 11-20 years, 68.4%; 21-30 years, 89.5%; 31 40 years, 94.7%; and > 41 years, 88.9%. Adults from the Indian subcontinent and Philippines had variable prevalence rates and Sri Lankans living in the region were highly serosusceptible (35%). Because of the clinical impact of varicella in adult populations, vaccine prevention might be beneficial. PMID- 15332756 TI - National study on the prevalence of iodine deficiency disorders among schoolchildren 8-12 years of age in Bahrain. AB - A cross-sectional survey of primary-school children in Bahrain was conducted to estimate the prevalence of goitre and iodine deficiency according to age, sex and area of residence. During January-May 1999, 1600 children were randomly chosen from all government schools. Children were examined for goitre and of those, 50% were randomly selected for urinary iodine level assessment. Only 26 children (1.7%) had goitre. Although median urinary iodine was above 100 microg/L, 121 of 749 children (16.2%) had low urinary iodine levels. Although iodine deficiency does not pose a significant public health problem in Bahrain, education about the nutritional value of iodized salts in the prevention of this disorder could increase public awareness. PMID- 15332757 TI - Study of cardiovascular disease risk factors among rural schoolchildren in Sousse, Tunisia. AB - We undertook an epidemiological survey based on a representative sample of 793 rural schoolchildren in Sousse, Tunisia to assess the prevalence of certain cardiovascular disease risk factors. The prevalence of hypertension (11.2%), hypercholesterolaemia (2.9%), hypertriglyceridaemia (1.0%), high levels of low density lipoprotein cholesterol (0.6%) and obesity (4.0%) showed no statistically significant difference based on sex. However, smoking (4%) showed a significant gender difference (boys: 7.3%; girls 1.2%). The relatively low cardiovascular disease risk factor profile of Tunisian children needs to be encouraged through to adulthood. Thus a school programme of heart health promotion should be established. PMID- 15332758 TI - Self-reported adolescents' health and gender: an Egyptian study. AB - To identify self-reported health problems among adolescents, a multistage, stratified random sample of 1002 adolescents from preparatory and secondary schools in Cairo (67.6% of the total sample) and the rural district of Qaliubia (32.4%) was surveyed. Of 863 completed questionnaires analysed, 54% were from males and 46% from females (age range: 12-18 years). The study showed that more boys than girls perceived their health as very healthy whereas more girls considered their health to be average. Significantly, more adolescent females reported weekly occurrence of abdominal pain, headache, dizziness, backache, morning tiredness, sleep disturbance and nervousness. Awareness-raising of parents and adolescents is necessary to allow determination of the relationship between gender and health. PMID- 15332759 TI - Gall bladder sludge and stones in multitransfused Egyptian thalassaemic patients. AB - One hundred Egyptian beta-thalassaemic patients on a long-term transfusion/chelation programme were evaluated for the prevalence of gall bladder sludge and stones and the associated risk factors. Fifty healthy individuals served as controls. Abdominal ultrasonography revealed that 14% of the thalassaemic patients had gall bladder sludge or stones (6% stones and 8% sludge). The thalassaemic patients with this complication were older, had a higher prevalence of gall bladder symptoms, higher levels of pretransfusion haemoglobin, larger amounts of transfused red cells, and more were regularly transfused. Multiple logistic regression analysis revealed that the presence that gall bladder symptoms and the amount of transfused red cells were the only significant predictors of the occurrence of gall bladder sludge or stones. PMID- 15332760 TI - Appendicitis in pregnancy. AB - The clinical files of 16,443 women delivered during the period June 1994 to June 2000 at Queen Alia Military Hospital were retrospectively studied for clinical presentation, investigation, operative findings and histopathological diagnosis of appendicitis during pregnancy. The result showed that 10 of the women underwent laparotomy for probable appendicitis. Of these, 8 had positive histopathological diagnosis. Appendicitis in pregnancy occurs infrequently. The clinical presentation varies and diagnosis is usually delayed. Right-side abdominal pain is the principal basis for diagnosis, while leukocytosis and low level fever, as in the non-pregnant state, are unreliable for diagnosis. PMID- 15332761 TI - Advanced maternal age and pregnancy outcome. AB - To assess the effect of maternal age on obstetric intervention and pregnancy outcome, a retrospective study compared obstetric intervention, pregnancy complications and outcome in 73 women of age > 35 years with 471 women of age 20 25 years attending Prince Ali Military Hospital, Jordan from June 1999 to May 2000. Older women were found to have significantly higher rates of medical complications such as hypertension and diabetes mellitus. Despite significantly increased frequency of large babies, trisomy 21, twin pregnancy and antepartum haemorrhage, overall outcome was satisfactory. We conclude that older women, managed by modern obstetric methods and delivered in a modern health-care centre, can expect good pregnancy outcomes. PMID- 15332762 TI - Iron status and socioeconomic determinants of the quantity and quality of dietary iron in a group of rural Iranian women. AB - Iron intake and status were investigated in 471 mothers (age range: 16-53 years) from rural areas in Khorramabad, Islamic Republic of Iran. Although average total iron intake was acceptable, only 6.4% of women derived at least 4% of their total intake from animal iron. Average energy and protein intakes were inadequate. Low iron status was seen in 8.2%-28.7%, depending on the parameter used, with 28.3% experiencing iron-deficiency anaemia. Significantly higher animal iron intakes were found in literate or employed women, or those of family size fewer than six people. Increasing employment opportunities, income levels and literacy rates for women will result in better iron intake and status and should receive particular attention in national planning. PMID- 15332763 TI - Physical fitness status among the students of a medical college in Kathmandu, Nepal. AB - The physical fitness of Nepalese students at a medical college in Kathmandu was measured. The proportions of students with poor fitness, average fitness, good fitness, very good fitness and excellent fitness were 10.4%, 23.6%, 39.6%, 17.9% and 8.5% respectively. No significant difference between the male and female students in physical fitness index score was noticed in any of the fitness level groups. PMID- 15332764 TI - Primary health care patients' knowledge about diabetes in the United Arab Emirates. AB - This study defined the baseline level of knowledge about diabetes and the perceived effectiveness of sources of diabetes information in an urban primary health care centre in Al-Ain. A validated questionnaire was administered to 300 randomly chosen adult patients with diabetes. Patient knowledge about diabetes was directly related to their level of general education. Written and electronic media, and contact with a nurse or doctor were effective sources of education on diabetes while "conversation with significant others", dietician or pharmacist were not effective. Nurse- or doctor-centred education appears to be an effective choice for future programmes to provide information to patients with diabetes. PMID- 15332765 TI - Low rates of diabetic patients reaching good control targets. AB - A cross-sectional study was conducted of 404 diabetic patients attending King Abdulaziz University Hospital medical clinic from June 1998 to January 2000 in order to determine their rates of reaching target levels for blood glucose, blood pressure, serum lipids and body mass index. Greatest difficulty was found in reaching target levels for low-density lipoprotein and glycated haemoglobin. Target levels for blood pressure were harder to achieve for Saudis than non Saudis. Significantly fewer females reached target levels for blood glucose, body mass index, low-density lipoprotein or all targets than males (both Saudi and non Saudi). Efforts are needed to improve compliance to diet and drug regimens and to identify and treat risk factors in each patient. PMID- 15332766 TI - TNF-alpha and homocysteine levels in type 1 diabetes mellitus. AB - The role of homocysteine as an independent risk factor for vascular endothelial damage, and the possible link between homocysteine and tumour necrosis factor alpha (TNF-alpha) as two synergistic risk factors for beta-cell apoptosis in type 1 diabetes mellitus was studied. Plasma homocysteine levels were significantly elevated in all diabetic patients compared with controls and diabetic patients with vascular complications showed higher elevations. Furthermore, homocysteine levels showed significant positive correlation with the degree of microalbuminuria. TNF-alpha levels were elevated in all diabetic patients compared with controls. These results may have therapeutic implications. PMID- 15332767 TI - Blood pressure control and target organ complications among hypertensive patients in southern Saudi Arabia. AB - Blood pressure (BP) control and target organ complications (TOC) among 203 hypertensive patients attending a hospital clinic in Abha, Saudi Arabia were studied. Optimal BP control was defined as BP < 140/90 and was achieved in 99 (48.8%) patients. Diuretics and calcium channel blockers were associated with the best rates of BP control while adverse drug reactions were associated with the poorest rates. TOC were classified according to World Health Organization criteria; 62% of patients had no TOC. Retinopathy, renal impairment and left ventricular hypertrophy were the most common stage II complications. Cerebrovascular accidents and ischaemic heart disease were the most common stage III complications. PMID- 15332768 TI - New biochemical markers in chronic heart failure. AB - We investigated the plasma levels of tumour necrosis factor-alpha (TNF-alpha), leptin and insulin, and their relation to body mass index (BMI) in 80 male patients who presented with chronic heart failure (mean age: 47 +/- 4 years) at Tanta University Hospital. Plasma leptin, TNF-alpha and insulin were significantly increased and BMI significantly decreased in New York Heart Association classes III and IV patients. TNF-alpha, leptin and insulin were positively correlated, and TNF-alpha and BMI and leptin and BMI were negatively correlated in stages III and IV of heart failure. We conclude that cytokine neuroendocrine activation may form part of advanced stage heart failure. It may also be responsible for worsening cachexia, and can be used as a marker to determine disease severity. PMID- 15332769 TI - Differential display analysis of mRNAs in chronic myelogenous leukaemia. AB - Analysis of expressed mRNAs with differential display-polymerase chain reaction (DD-PCR) is a powerful tool for the characterization of genes involved in malignant pathways and might identify markers for different phases of chronic myelogenous leukaemia (CML). We examined the presence of BCR-ABL transcripts in 25 CML patients in either the chronic phase or blast crisis. We then analysed the expression of leukocytic RNA transcripts in CML phases. DD-PCR technique was used to examine CML cases with BCR-ABL in comparison with CML cases lacking detectable BCR-ABL transcripts. Our results support the use of differential display not only for characterization of the CML differentially expressed genes but also to locate patterns that can be implemented as valuable fingerprints for each phase of CML. PMID- 15332770 TI - Central obesity in elderly individuals in south-western Saudi Arabia: prevalence and associated morbidity. AB - Central obesity in all individuals aged 65 years and over (n = 810) in the catchment areas of three primary health care centres in Abha was determined from the waist circumference (WC) and waist-to-hip ratio (WHR). The age-adjusted prevalence of central obesity was 32.4% and 43.5% based on the WC and WHR indicators respectively. WC was significantly associated with the risk of diabetes and hypertension, while WHR was significantly associated with the risk of diabetes only. These findings suggest that reducing the prevalence of central obesity in old age would decrease the risk of diabetes and hypertension. WC is a powerful independent predictor mainly of hypertension risk, while WHC is a good predictor of the risk of diabetes. PMID- 15332771 TI - Incidence of hip fractures in the Lebanese population. AB - This is the first prospective study to estimate the incidence of hip fractures in the general population in Lebanon. All orthopaedic surgeons in Beirut reported on hip fracture cases between 1 October and 31 December 1996. Data were extrapolated to 1 year and standardized for the whole population. The estimated annual incidence rate was 0.129% (women: 0.153%, men: 0.100%), increasing with age and remaining higher in women. Our figures are comparable with larger studies in other Mediterranean countries. The incidence rate of hip fractures in our population falls within a gradient of risk between the west and the east and the north and the south, highlighting the importance of environmental and genetic factors as risk factors for osteoporotic fractures. PMID- 15332772 TI - Bone density estimates and risk factors for osteoporosis in young women. AB - The bone mineral density (BMD) of the calcaneus was estimated in 185 young women from the United Arab Emirates, using SAHARA ultrasound. All participants completed a questionnaire on factors potentially associated with osteoporosis. In all, 29 (15.7%) of the women were classified as having osteopenia and none as having osteoporosis. Participants with osteopenia were more likely to have had a later onset of menarche, irregular periods, lower body mass index, and a positive family history of osteoporosis. Only late menarche and low body mass index, however, were independent predictors of osteopenia. PMID- 15332773 TI - Infection following orthopaedic implants and bone surgery. AB - Forty-seven patients were investigated for early or late postoperative infections of orthopaedic implants and/or bone. A total of 88 isolates were recovered (64 aerobes and 24 anaerobes). Pseudomonas aeruginosa and Staphylococcus epidermidis were the most common causative agents. Anaerobic bacteria were isolated from 16 (34%) patients; 50% of patients with late-onset infection and 10.5% with early onset infection. In 6 (12.8%) patients, infection was with anaerobic organisms alone. All these patients had retained an extramedullary internal fixation device. Anaerobic microorganisms appear to play a significant role in the pathogenesis of late-onset postoperative infection, especially where there is an extramedullary internal fixation device. PMID- 15332774 TI - Multicentre surveillance of antibiotic resistance in nosocomial Staphylococcus aureus in Cyprus. AB - Four general hospitals in Cyprus evaluated the incidence of methicillin-resistant Staphylococcus aureus (MRSA) strains in positive cultures of staphylococci among inpatients from September 1999 to March 2000. One hundred and ninety three (193) strains were isolated from cultures of respiratory secretions, skin and mucous membrane lesions, blood and urine. Of the S. aureus strains studied, 80 (41.45%) were MRSA and 113 (58.55%) were methicillin sensitive S. aureus (MSSA). More than 78.75% of the MRSA were resistant to erythromycin and 18.75% to gentamicin. Of the MSSA, 17.69% were resistant to erythromycin and 7.08% to gentamicin. None of the MRSA and MSSA strains showed reduced sensitivity to vancomycin. PMID- 15332775 TI - Prevalence of Clostridium difficile-associated diarrhoea among hospitalized Jordanian patients. AB - We investigated stool specimens of 400 patients at Jordan University Hospital (300 patients with clinical diarrhoea and 100 controls without diarrhoea) for the presence of Clostridium difficile or its toxin. We found a 9.7% prevalence rate of C. difficile or its toxin in stools of patients with diarrhoea. The prevalence of other potential enteric pathogens, such as Salmonella spp. (2.3%), Shigella spp. (1.0%) and Entamoeba histolytica (2.7%), was significantly less. Prevalence of C. difficile or its toxin in controls was 3.0%. Toxin A was detected in 93.1% of C. difficile-associated diarrhoea cases using an enzyme immunoassay. Our study indicates that C. difficile-associated diarrhoea is mostly observed among hospitalized patients aged > or = 50 years, in association with antimicrobial treatment. PMID- 15332776 TI - Detection and characterization of verotoxin-producing strains among sorbitol non fermenting Escherichia coli. AB - The presence of genes for verotoxin 1 and 2 (VT1 and 2) among sorbitol non fermenting Escherichia coli isolates from diarrhoeal cases was assessed using polymerase chain reaction assay. Of 60 (88%) positive isolates, 37 (62%) harboured VT1 and 23 (38%) both VT1 and VT2. In HeLa cell adherence assay, 48 (71%) isolates exhibited mannose-resistant adherence to HeLa cells. Multidrug resistance was observed in 56 (82%) isolates, with ampicillin, chloramphenicol, streptomycin, sulfamethoxazole-trimethoprim and tetracycline pattern being the most common. There were 13 common and 22 single biochemical phenotypes identified. Isolates belonging to common biochemical phenotypes normally had a similar pattern of adherence and VT production, but differed greatly in their pattern of antibiotic resistance, pointing to a high rate of antibiotic resistance transfer among these isolates. PMID- 15332777 TI - Urinary calculi: bacteriological and chemical association. AB - We investigated the bacteriology of urinary calculi in relation to urinary tract infection, stone formation, chemical composition and antibiotic sensitivity. Fifty-two patients (37 males, 15 females) with urolithiasis were studied. Urine, serum and urinary calculi specimens were taken and serum biochemical tests to detect uric acid, calcium and phosphorus were performed. Urine analysis and culture were also performed. Of the 52 patients, 19 (37%) had associated urinary tract infection, with Escherichia coli and Proteus mirabilis being the most common causative microorganisms. The bacterial isolates from urine and those from calculi differed in their susceptibility to antimicrobial agents. We conclude that in over 50% of patients with urolithiasis, urine culture can detect the infecting organisms associated with stone formation and the organisms within urinary calculi. PMID- 15332778 TI - Prevalence of hepatitis C virus among patients with liver disease in the Republic of Yemen. AB - We investigated 143 patients with various classes of liver disease and 120 healthy subjects for serological markers of hepatitis C and B viruses. We found a prevalence rate of 37.1% of anti-HCV markers in patients with liver disease (cases), and 4.2% and in the control group. HBsAg was detected in 33.6% of cases and 13.3% of the controls. Anti-HCV and HBsAg were detected in 7.7% of cases but were not detected in the controls. Anti-HCV, HBsAg, anti-HBe and anti-HDV were detected in three patients. Serum protein electrophoresis results showed elevated gammaglobulin and low albumin in patients positive for anti-HCV who had liver disease. PMID- 15332779 TI - Effect of interferon therapy on fatty acid saturation index in hepatitis C virus infection. AB - The ratio of stearic to oleic acids, i.e. the fatty acid saturation index, in red blood cell membranes was assayed in 60 patients with chronic hepatitis C virus infection before and after interferon-alpha therapy. Results were compared with 20 healthy controls. Hepatitis C virus titre was also assayed before and after interferon-alpha therapy. Within 2-5 months following interferon-alpha therapy, a significant inverse correlation was observed between saturation index and hepatitis C virus load. We conclude that hepatitis C virus infection enhances the degree of desaturation of 18-carbon fatty acids and that interferon-alpha is involved in their metabolism by increasing the degree of saturation and subsequent decrease in membrane fluidity. PMID- 15332780 TI - Efficacy of albendazole in giardiasis. AB - Albendazole and metronidazole were compared in 68 patients diagnosed positive for giardiasis. Albendazole 1200 mg, one dose was given to 24 patients, albendazole 400 mg twice a day for 3 days was given to 23 patients, and metronidazole 400 mg 3 times a day for 5 days to 21 patients. Response to therapy was monitored by clinical examination and analysis of fresh faecal samples on days 0, 3, 7 and 10. Response to the single dose of albendazole was 55%, to the divided dose of albendazole 70%, and to metronidazole 84%. The results show that albendazole, originally recommended for helminthic infection, can also be used in patients with mixed protozoal infection or for infections resistant to metronidazole. PMID- 15332783 TI - [Availability and use of emergency services in Tunisia: principal results of an exhaustive national survey]. AB - In order to assess hospital emergency rooms, a comprehensive national epidemiological investigation was conducted in all 155 public emergency structures in Tunisia. Here we present the main results of the levels of availability and use of emergency services. Coverage of the population by services is adequate (one emergency service per 60,000 people). Emergency wards admit about 2,500,000 patients every year (a quarter of the population of the country). The university hospital emergency services are the most heavily used (150 patients per day on average). Emergency services are sought for medical (60%), surgical (18%), paediatric (14%) and gynaecological reasons (5%). It would be useful to assess the quality of care delivered and the satisfaction of citizens and health workers. PMID- 15332781 TI - An epidemiological profile of brucellosis in Tabuk Province, Saudi Arabia. AB - All 137 brucellosis cases occurring in Tabuk Province, Saudi Arabia in 1997 were studied retrospectively. Brucella agglutination titre of > or = 1/80, or rising titre plus history of typical signs and symptoms were considered evidence of infection. The incidence rate was 34/100,000, mean age 33.8 +/- 13.9 years (range: 3-72 years) and male:female ratio 1.8:1. There were 63.5% of cases rurally resident, 58.4% kept animals at home or elsewhere, 27.0% worked with animals and/or on farms, and 88.3% reported a history of raw milk ingestion. The most common infecting agents were Brucella melitensis, B. abortus and B. suis. Splenomegaly and hepatomegaly were detected in 25.5% and 22.6% of cases respectively. PMID- 15332782 TI - The role of vitamin A in enhancing humoral immunity produced by antirabies vaccine. AB - We tested the effects of vitamin A supplementation on the antibody titre of 40 healthy volunteers (age range: 10-35 years), who had received a complete course of antirabies vaccine (5 injections over 30 days). After determining the baseline serum vitamin A status of 80 volunteers, 20 pairs were matched for serum vitamin A level, body mass index, age, sex and socioeconomic status. One person from each pair was randomly assigned to an experimental or control group. The experimental group received vitamin A and antirabies vaccine. Controls received only the vaccine. The experimental group had significantly greater (2.1 times) serum antirabies titre than controls. This finding is an important step towards improving the economy of dosage of antirabies vaccines. PMID- 15332784 TI - Impact of triage in accident and emergency departments in Bahrain. AB - We aimed to assess the impact of triage by physicians on the workload and expenditure of the Accident and Emergency (AE) Department of Salmaniya Medical Complex, Bahrain. We analysed three sets of data: patient visits to the AE Department over a 9-month period; patient visits 1 year previously; and forecast patient visits over 9 months starting from July 1999. The referral of patients to AE cubicles was reduced by 54.4% after the implementation of the triage, and reduction in the workload was statistically significant. The reduction in health care expenditure was estimated at between 15.3% and 17.3%. We conclude that triage by physicians can be cost-effective and can reduce the AE Department workload, freeing more time to manage life-threatening and urgent cases. PMID- 15332785 TI - Prevalence of tobacco use among the adult Lebanese population. AB - To determine the prevalence of smoking in Lebanon, 727 individuals aged > or = 19 years were randomly selected for study using Emile Roux and Fagerstrom questionnaires. Smokers were defined as those answering "yes" to the question, "Do you currently smoke?" A Fagerstrom score > or = 6 indicated strong nicotine addiction. The prevalence of smoking was 53.6%. The male/female ratio was 1.23, with 67.0% of smokers categorized as addicted. Failure to quit was related to withdrawal symptoms, mostly irritability (57%) and weight gain (20%). Recommendations are given for combating this high prevalence of tobacco use. PMID- 15332786 TI - Measles immunization in Saudi Arabia: the need for change. AB - This paper describes the measles immunization programme in Saudi Arabia and the change from the single-dose schedule with the Schwartz vaccine to the double-dose schedule with the Edmonston-Zagreb vaccine. The recent measles-mumps-rubella school campaign is also described. PMID- 15332787 TI - Bone marrow transplantation in thalassaemia patients in Shiraz, Islamic Republic of Iran. AB - We report the results of allogeneic bone marrow transplantation in 26 female and 37 male patients with beta-thalassaemia major (age range: 2-17 years), performed at Namazi Hospital over the period 1992-99. In all cases, standard conditioning and immunosuppressive regimens were employed. Currently, 50 patients remain thalassaemia-free, 9 of whom have developed chronic graft-versus-host disease. There were 8 deaths among the 13 unsuccessful transplant cases: 4 due to acute uncontrollable graft-versus-host disease, and 4 to non-transplant-related causes such as hypoglycaemia, hypersensitivity reactions and advanced disease. We conclude that allogeneic bone marrow transplantation is an effective therapy for the treatment of beta-thalassaemia major, particularly for patients classified as classes I and II. PMID- 15332788 TI - Laparoscopic cholecystectomy in Jordan. AB - This study was conducted on 100 patients with symptomatic gallbladder stones, aged 22-81 years with a mean of 51.5 years, who underwent cholecystectomy in Zarqa city, Jordan between July 1998 and July 1999. The success rate was 87% and the procedure was completed using the conventional method in 13 patients. The mean operative time was 60 minutes, complication rate was 5% and there were no deaths. The mean hospital stay was 1 day and mean time to return to work was 10 days. This study showed that laparoscopic cholecystectomy is a safe procedure with reasonable operative time, less postoperative pain, a short hospital stay, early return to work, and a low morbidity and mortality rate. PMID- 15332789 TI - High breast milk sodium concentration resulting in neonatal hypernatraemic dehydration. PMID- 15332790 TI - Amniotic constriction band sequence. PMID- 15332791 TI - Imaging of tumoural calcinosis. PMID- 15332792 TI - Bilateral pleural empyema following periodontal abscess. PMID- 15332793 TI - Spectroscopic evidence for the direct coordination of the pertechnetate anion to the uranyl cation in [UO2(TcO4)(DPPMO2)2]+. AB - We report the synthesis and structural characterization of [UO(2)(ReO(4))(DPPMO(2))(2)][ReO(4)] and [UO(2)(Cl)(DPPMO(2))(2)][Cl] (where DPPMO(2) = bis(diphenylphosphino)methane dioxide). In both complexes, the linear uranyl dication is coordinated to two bidentate DPPMO(2) ligands in the equatorial plane with one coordinated and one non-coordinated anion (either perrhenate or chloride). We have also prepared the pertechnetate analogue, and, through (31)P and (99)Tc NMR, we have shown that the cation, [UO(2)(TcO(4))(DPPMO(2))(2)](+), is stable in solution. PMID- 15332794 TI - Feasibility of a two-metal, four-purine nucleobase quartet motif. AB - A mixed-purine nucleobase complex of composition trans-[(NH(3))(2)Pt(9-EtA-N1)(9 MeHx-N7)](NO(3))(2).2H(2)O (1) (9-EtA = 9-ethyladenine; 9-MeHx = 9 methylhypoxanthine) has been prepared and characterized by X-ray crystallography. Cations of 1 are self-complementary as far as hydrogen bonding properties are concerned and form H bonded dimers, containing four intermolecular hydrogen bonds in addition to two intramolecular ones. The resulting mixed-purine square is considered a model compound for a putative mixed-purine tetrad consisting of two adenines and two guanines. In this model, the one-metal, four-nucleobase quartet motif, as seen in guanine or uracil quartets of nucleic acids, with the metal located in the center of the base tetrad, has been altered to a two-metal, four nucleobase motif, with the two metal ions localized at the periphery. PMID- 15332795 TI - Linear NiII-MnIII2-NiII tetramers: an oligomeric component of the MnIII2NiII single-chain magnets. AB - Heterometallic linear tetramers [Mn(5-R-saltmen)Ni(pao)(bpy)(2)](2)(ClO(4))(4) (5 R-saltmen(2-) = N,N'-1,1,2,2-tetramethylethylene bis(5-R-salicylideneiminate); pao(-) = pyridine-2-aldoximate; bpy = 2,2'-bipyridine, R = H, 1; Cl, 2; Br, 3; MeO, 4) have been synthesized and structurally characterized. These compounds exhibit a [Ni(II)-NO-Mn(III)-(O)(2)-Mn(III)-ON-Ni(II)] skeleton where -ON- is an oximate bridge between Mn(III) and Ni(II) ions and -(O)(2)- is a bi-phenolate bridge between Mn(III) ions. These tetramers can be seen as oligomeric units of the heterometallic Mn(III)(2)-Ni(II) chain observed in a family of single-chain magnets (Clerac, R.; Miyasaka, H.; Yamashita, M.; Coulon, C. J. Am. Chem. Soc. 2002, 124, 12837. Miyasaka, H.; Clerac, R.; Mizushima, K.; Sugiura, K.; Yamashita, M.; Wernsdorfer, W.; Coulon, C. Inorg. Chem. 2003, 42, 8203.). Magnetic measurements on these tetramers confirm the nature of the magnetic interactions reported for the Mn(III)(2)-Ni(II) chains: a strong antiferromagnetic Mn(III)/Ni(II) coupling via the oximate bridge (J(Ni-Mn) ranges from -23.7 to -26.1 K) and a weak ferromagnetic Mn(III)/Mn(III) coupling through the bi-phenolate bridge (J(Mn-Mn) ranges from +0.4 to +0.9 K). These magnetic interactions lead to tetramers with an S = 2 ground state. PMID- 15332796 TI - A new route to obtain shape-controlled gold nanoparticles from Au(III)-beta diketonates. AB - Ligands with a beta-diketone skeleton have been employed for the first time as reductant to produce ligand stabilized gold nanoparticles of different shapes from aqueous HAuCl(4) solution. Evolution of stable gold nanoparticles follows first order (k approximately equal to 10(-2) min(-1)) kinetics with respect to Au(0) concentration. Growth of particles of different shapes (spherical or triangular or hexagonal) goes hand in hand under the influence of different beta diketones, which have excellent capping and reducing properties. Chlorine insertion was observed to take place in the beta-diketone skeleton. PMID- 15332797 TI - Gadolinium(III) 1,2-hydroxypyridonate-based complexes: toward MRI contrast agents of high relaxivity. AB - Prospective gadolinium(III) MRI contrast agent precursors [Gd-TREN-1,2-HOPO] (1) [TREN-1,2-HOPO = tris[(1-hydroxy-2-oxo-1,2-dihydropyridine-6 carboxamido)ethyl]amine] and [Gd-TREN-bis(Me-3,2-HOPO)-1,2-HOPO] (2) have been synthesized and characterized by relaxometric measurements. The water proton relaxivity values of 1 and 2 (20 MHz and 25 degrees C) are 9.5 and 9.3 mM(-)(1)s( )(1), respectively, suggesting the presence of two coordinated water molecules. The molecular structure of [1.DMF](2) was obtained and reveals a similar eight coordinate geometry to [Gd-TREN-Me-3,2-HOPO.2H(2)O] ([3.2H(2)O]). A shape analysis of the coordination polyhedron of 1 reveals that this geometry is best described as a bicapped trigonal prism, poised to accommodate an additional donor atom to give a tricapped trigonal prismatic intermediate. This geometry supports the model that formation of a tris-aquo intermediate for 1 enables fast and associative water exchange. PMID- 15332798 TI - Reactivity of pyridine-2,4,6-tricarboxylic acid toward Zn(II) salts under different reaction conditions. AB - Pyridine-2,4,6-tricarboxylic acid (ptcH(3)) readily reacts with a Zn(II) salt at room temperature to form different products depending upon the presence or absence of pyridine in the reaction mixture. In the presence of pyridine, the ligand breaks to form infinitely zigzag coordination polymers with the empirical formula [Zn(Ox)(py)(2)]n(Ox = oxalate, py = pyridine). The backbone is formed from Zn(II)-oxalate where two pyridine molecules are coordinated to each Zn(II) ion giving it hexacoordination. The orientation of the bound pyridines is slightly different when Zn(II)-nitrate is used compared to that when Zn(II) sulfate (or acetate) salt is used. In absence of pyridine, the ligand remains intact and forms a mixture of a carboxylate-bridged coordination polymer and a discrete carboxylate-bridged 12-membered metallomacrocycle. PMID- 15332799 TI - Upward temperature shift of the intrinsic phase lag of the magnetization of Bis(phthalocyaninato)terbium by ligand oxidation creating an S = 1/2 spin. AB - An alternating-current (ac) magnetic susceptibility measurement for the [(Pc)(2)Tb(III)](0) complex (Pc = phthalocyaninato) has shown that ligand oxidation of the anionic [(Pc)(2)Tb(III)](-) complex gives rise to a significant upward shift of the temperature range where the magnetization response shows a phase lag behind the time-varying external magnetic field. The peaks of the out of-phase component of the ac susceptibility of the pi-radical [(Pc)(2)Tb(III)](0) were observed at 50, 43, and 36 K with ac magnetic fields of 10(3), 10(2), and 10 Hz, respectively, which were more than 10 K higher than the corresponding values of the anionic complex with a closed-shell pi-system. The ac susceptibility measurements on the complex with octa(dodecoxy)-substituted Pc ligand, which is readily dilutable in diamagnetic media, proved that the significant rise of the temperatures occurs as an intrinsic single-molecular property of the complex possessing both J = 6 and S = (1)/(2) systems, and is not due to long-range magnetic order or interactions between adjacent unpaired pi-electrons. PMID- 15332800 TI - Synthesis, structure, and bonding of the nonclassical Zintl phase K5As3Pb3. AB - The title phase was synthesized by direct fusion of a stoichiometric amount of the elements followed by annealing at 650 degrees C for 3 weeks. The compound crystallizes in the orthorhombic space group Pnma (No. 62), Z = 4, with a = 19.451(6) A, b = 12.164(3) A, c = 6.581(1) A. The compound is made up of As(3)Pb(3)(5-) crown clusters that can be likened geometrically and electronically to 6-atom hypho-clusters derived from a tricapped trigonal prismatic closo parent. These crowns are interconnected via intercluster Pb-Pb bonds to form infinite chains along the b axis, which means the compound contains an extra two cations and two electrons per formula unit. Extended Huckel calculations indicate that the two additional electrons per cluster are accommodated in pi states on the cluster and predict that the phase should be semiconducting. The latter is confirmed by microwave resistivity measurements, rho(298) = 1.0 x 10(2) microOmega.cm; (deltarho/deltaT)/rho = -0.14(3) K(-)(1). PMID- 15332801 TI - Large spin differences in structurally related Fe6 molecular clusters and their magnetostructural explanation. AB - The syntheses, crystal structures, and magnetic characterizations of three new hexanuclear iron(III) compounds are reported. Known [Fe(6)O(2)(OH)(2)(O(2)CBu(t))(10)(hep)(2)] (1) is converted to new [Fe(6)O(2)(OH)(O(2)CBu(t))(9)(hep)(4)] (3) when treated with an excess of 2-(2 hydroxyethyl)-pyridine (hepH). Similarly, the new compound [Fe(6)O(2)(OH)(2)(O(2)CPh)(10)(hep)(2)] (2), obtained from the reaction of [Fe(3)O(O(2)CPh)(6)(H(2)O)(3)] with hepH, is converted to [Fe(6)O(2)(OH)(O(2)CPh)(9)(hep)(4)] (4) when treated with an excess of hepH. This can be reversed by recrystallization from MeCN. The cores of the four Fe(6) complexes all comprise two triangular [Fe(3)(mu(3)-O)(O(2)CR)(3)(hep)](+3) units connected at two of their apices by two sets of bridging ligands. However, 1 and 2 differ slightly from 3 and 4 in the precise way the two Fe(3) units are linked together. In 1 and 2, the two sets of bridging ligands are identical, consisting of one mu-hydroxo and two mu-carboxylate groups bridging each Fe(2) pair, i.e., a (mu-OH(-))(mu-O(2)CR(-))(2) set. In contrast, 3 and 4 have two different sets of bridging ligands, a (mu-OH(-))(mu-O(2)CR(-))(2) set as in 1 and 2, and a (mu-OR( ))(2)(mu-O(2)CR(-)) set, where RO(-) refers to the alkoxide arm of the hep(-) chelate. Variable-field and -temperature dc magnetization measurements establish that 1 and 2 have S = 5 ground states and significant and positive zero-field splitting parameters (D), whereas 3 and 4 have S = 0 ground states. This dramatic difference of 10 unpaired electrons in the ground state S values for near isomeric compounds demonstrates an acute sensitivity of the magnetic properties to small structural changes. The factors leading to this have been quantitatively analyzed. The semiempirical method ZILSH, based on unrestricted molecular orbital calculations, was used to obtain initial estimates of the Fe(2) pairwise exchange interaction constants (J). These calculated values were then improved by fitting the experimental susceptibility versus T data, using a genetic algorithm approach. The final J values were then employed to rationalize the observed magnetic properties as a function of the core topologies and the presence of spin frustration effects. The large difference in ground state spin value was identified as resulting from a single structural difference between the two types of complexes, the different relative dispositions (cis vs trans) of two frustrated exchange pathways. In addition, use of the structural information and corresponding J values allowed a magnetostructural correlation to be established between the J values and both the Fe-O bond distances and the Fe-O-Fe angles at the bridging ligands. PMID- 15332802 TI - Dramatic structural effects of a single hydrogen atom in HNPBu t 3. AB - The molecular structure of tri-tert-butylphosphine imide has been re-determined using the recently developed DYNAMITE method, which allows all assumptions about local symmetry to be removed without increasing the number of refining structural parameters excessively. The imide hydrogen causes the NPBu(t)(3) group to deviate hugely from local C(3) symmetry, with N-P-C angles returned as 99.2(9), 110.9(7), and 111.5(11) degrees, while the C-P-C angles also deviate from symmetry, being 109.8(8), 110.5(9), and 113.9(9) degrees, so that the NPC(3) fragment is close to C(s) rather than C(3) symmetry. The application of the DYNAMITE method to HNPBu(t)(3) also allows the methyl groups to be asymmetric, which has been shown to be important by ab initio methods. The re-determination of this structure using these more sophisticated methods has also resulted in a much shorter P-N bond than was previously determined, and is consistent with the molecule being regarded as HN=PBu(t)(3), rather than HN(-)-P(+)Bu(t)(3). PMID- 15332803 TI - Synthesis of and structural studies on lead(II) cysteamin complexes. AB - The novel compounds PbCl(2).(SCH(2)CH(2)NH(3)) (1), Pb(SCH(2)CH(2)NH(2))(2).2PbCl(SCH(2)CH(2)NH(2)) (2), and Pb(SCH(2)CH(2)NH(2))(2) (3) were synthesized by reaction of PbO or PbCl(2) with [HSCH(2)CH(2)NH(3)]Cl and NaOH, and were characterized by elemental analysis, IR-, and UV/vis-spectroscopy. Single-crystal X-ray diffraction revealed different coordination modes for the two Pb atoms in 2. The Pb atom in the Pb(SCH(2)CH(2)NH(2))(2) unit forms two covalent Pb-S and two intramolecular dative Pb...N bonds, leading to a pseudo trigonal bipyramidal configuration with a stereochemically active lone pair. The Pb atom in the PbCl(SCH(2)CH(2)NH(2)) unit, the first moiety structurally characterized of the PbCl(SR) type (R = organic group), forms covalent Pb-Cl and Pb-S bonds, an intramolecular dative Pb...N bond, and two intermolecular Pb...S contacts, giving a pseudo octahedral configuration with a stereochemically active lone pair as well. Despite the Pb(SCH(2)CH(2)NH(2))(2) moiety exhibiting C(2) symmetry in 2, and C(1) symmetry in 3, its structural parameters are rather similar in the two compounds. The influence of the Pb...N bond on molecular structure and thermodynamic stability were estimated by means of quantum chemical ab initio methods. Although an analysis of the wave function in terms of natural bond orbitals (NBO) revealed that n(N) and n(p)(S) compete for the empty p orbital of the Pb(II) atom, the sigma-type n(N)-6p(Pb) interaction is stronger than the pi-type n(p)(S)-6p(Pb) interaction and hence determines the conformation of the compounds. PMID- 15332804 TI - Synthesis and characterization of ReO4-containing microporous and open framework structures. AB - A microporous and an open framework structure, [Cu(2)(pzc)(2)(H(2)O)(2)ReO(4)] (I) and [Cu(pzc)(H(2)O)ReO(4)].2H(2)O (II) (pzc = 2-pyrazinecarboxylate), respectively, have been prepared using hydrothermal methods and characterized using IR, TGA, and X-ray diffraction (I Pnma, No. 62, Z = 4, a = 7.4949(9) A, b = 24.975(3) A, c = 9.141(1) A; II P2(1)/c, No. 14, Z = 4, a = 8.5878(9) A, b = 12.920(1) A, c = 9.741(1) A, beta = 92.830(2) degrees ). I and II crystallize as red and blue solids, respectively, and each contains chains constructed from alternating Cu(pzc)(2)/ReO(4) oxide-bridged metal sites. The bidentate pzc ligand further bridges each -Cu-O-Re-O- chain to adjacent chains, via the Cu sites, to form a 3D net in I, with ellipsoidal channels that are approximately 3.3-4.7 A x 12.5 A, and in II, stacked layers of square nets with H(2)O-filled cavities that are approximately 4.4 x 5.1 A. Local ReO(4)(-) groups, a component of common oxidation catalysts, are directed at the channels and cavities of each structure, respectively. Thermogravimetric analysis indicates that I loses up to 64% of its H(2)O content before decomposition at 225 degrees C, while II loses approximately 100% of its H(2)O content by 265 degrees C. PMID- 15332805 TI - Synthesis and photochemical properties of a novel iron-sulfur-nitrosyl cluster derivatized with the pendant chromophore protoporphyrin IX. AB - The novel Roussin red-salt ester (PPIX-RSE) with a pendant porphyrin chromophore was prepared and investigated as a precursor for the photochemical generation of nitric oxide. PPIX-RSE has the general formula Fe(2)(NO)(4)[(mu-S,mu-S')P] (where (S,S')P is the bis(2-thiolatoethyl) diester of protoporphyrin IX. The photoexcitation of PPIX-RSE with 436- or 546-nm light in an aerated chloroform solution led to the photodecomposition of the cluster with the respective quantum yields (5.2 +/- 0.7) x 10(-4) and (2.5 +/- 0.5 x 10(-4)) and the concomitant release of NO. PPIX-RSE is a significantly more effective NO generator at longer wavelength excitation than are other Fe(2)(mu-SR)(2)(NO)(4) esters for which R is a simple alkyl group such as CH(3)CH(2)- because of the much higher absorptivity of the pendant PPIX chromophore at these wavelengths and a modestly higher quantum yield. Fluorescence intensity and lifetime data indicate that the photoexcited porphyrin of PPIX-RSE is largely quenched by the energy transfer to the Fe(2)S(2)(NO)(4) cluster's core. However, a small fraction of this emission is not quenched, and it is proposed that PPIX-RSE may exist in solution as two conformers. PMID- 15332806 TI - Self-assembly using dynamic coordination chemistry and hydrogen bonding: mercury(II) macrocycles, polymers and sheets. AB - The self-assembly of extended metal-containing arrays is described based on dynamic coordination chemistry at mercury(II) with bis(amidopyridyl) ligands to form macrocycles, polymers, or sheets which can be further organized by hydrogen bonding between amide substituents. The ligands 1,2-C6H4[NHC(O)-4-C5H4N]2, 1, 1,2 C(6)H(4)[C(O)NHCH(2)-4-C(5)H(4)N](2), 2, and 1,2-C(6)H(4)[CH(2)C(O)NHCH(2)-4 C(5)H(4)N]2, 3 can adopt polar conformations and so can confer helicity in their complexes. Several macrocycles of formula [(HgX(2))(2)(micro-LL)(2)] (LL = 1, 2), with tetrahedral mercury(II) centers, were prepared in which individual molecules are further self-assembled via hydrogen bonding in the solid state to form one- or two-dimensional polymers or sheets. In one case, a one-dimensional polymer [((HgX2)-(mu-3))n] was formed. It is shown that the mercury(II) centers can be six-coordinate in forming the sheet structure [((HgX2)(mu-2)2)n], in which there are particularly large pores. PMID- 15332807 TI - Powder diffraction study of a coordination polymer comprised of rigid building blocks: [Rh2(O2CCH3)4.mu2-Se2C5H8-Se,Se']infinity. AB - The crystal structure of a new hybrid product comprised of two rigid building blocks, namely dirhodium(II) tetraacetate, [Rh(2)(O(2)CCH(3))(4)] (1), and 2,6 diselenaspiro[3.3]heptane, Se(2)C(5)H(8) (2), has been solved ab initio using laboratory source X-ray powder diffraction (XRPD) data. The rigid body refinement approach has been applied to assist in finding an adequate model and to reduce the number of the refined parameters. Complex [Rh(2)(O(2)CCH(3))(4).mu(2) Se(2)C(5)H(8)-Se,Se'] (3) conforms to the triclinic unit cell with lattice parameters of a = 8.1357(4), b = 8.7736(4), and c = 15.2183(8) A, alpha = 77.417(3), beta = 88.837(3), and gamma = 69.276(4) degrees, V = 989.66(8) A(3), and Z = 2. The centrosymmetric P space group was selected for calculations. The final values of the reduced wR(p), R(p), and chi(2) were calculated at 0.0579, 0.0433, and 5.95, respectively. The structure of 3 is a one-dimensional zigzag polymer built on axial Rh...Se interactions at 2.632(6) A. The 2,6 diselenaspiro[3.3]heptane ligand acts as a bidentate linker bridging dirhodium units via both selenium atoms. The geometrical parameters of individual groups for rigid body refinement have been obtained from X-ray powder data for dirhodium(II) tetraacetate (1) and from single-crystal X-ray diffraction for diselenium molecule 2. The crystal structures of 1 and 2 are reported here for the first time. For 1 indexing based on XRPD data has resulted in the triclinic unit cell P with lattice parameters of a = 8.3392(7), b = 5.2216(5), and c = 7.5264(6) A, alpha = 95.547(10), beta = 78.101(6), and gamma = 104.714(13) degrees, V = 309.51(5) A(3), and Z = 1. The final values were wR(p) = 0.0452, R(p) = 0.0340, and chi(2) = 1.99. The 1D polymeric motif built on axial Rh.O interactions of the centrosymmetric dirhodium units has been confirmed for the solid-state structure of 1. Compound 2,6-diselenaspiro[3.3]heptane (2) conforms to the monoclinic space group P2(1)/c with the unit cell parameters of a = 5.9123(4), b = 19.6400(13), and c = 5.8877(4) A, beta = 108.5500(10) degrees, V = 648.15(8) A(3), and Z = 4. PMID- 15332808 TI - Reactivity of iron(II) 5,10,15,20-tetraaryl-21-oxaporphyrin with arylmagnesium bromide: formation of paramagnetic six-coordinate complexes with two axial aryl groups. AB - Coordination of sigma-aryl carbanions by chloroiron(II) 5,20-ditolyl-10,15 diphenyl-21-oxaporphyrin (ODTDPP)Fe(II)Cl has been followed by (1)H NMR spectroscopy. Addition of pentafluorophenyl Grignard reagent (C(6)F(5))MgBr to the toluene solution of (ODTDPP)Fe(II)Cl in the absence of dioxygen at 205 K resulted in the formation of the high-spin (ODTDPP)Fe(II)(C(6)F(5)). The titration of (ODTDPP)Fe(II)Cl with a solution of (C(6)H(5))MgBr carried at 205 K yields a rare six-coordinate species which binds two sigma-aryl ligands [(ODTDPP)Fe(II)(C(6)H(5))(2)](-). Warming of the [(ODTDPP)Fe(II)(C(6)H(5))(2)](-) solution above 270 K results in the decomposition to mono-sigma-phenyliron species (ODTDPP)Fe(II)(C(6)H(5)). Controlled oxidation of [(ODTDPP)Fe(II)(C(6)H(5))(2)](-) with Br(2) affords (ODTDPP)Fe(III)(C(6)H(5))Br, which demonstrates a typical (1)H NMR pattern of low-spin sigma-aryl iron(III) porphyrin. The considered oxidation mechanism involves the (ODTDPP)Fe(III)(C(6)H(5))(2) species, which is readily reduced to the iron(I) 21 oxaporphyrin, followed by oxidation with Br(2) and replacement of one bromide anion by aryl substituent. The (1)H NMR spectra of paramagnetic iron complexes have been examined in detail. Functional group assignments have been made with the use of selective deuteration. The peculiar (1)H NMR spectral features of [(ODTDPP)Fe(II)(p-CH(3)C(6)H(4))(2)](-) (sigma-p-tolyl: ortho, 30.8; meta, 53.6; para-CH(3), 42.1; furan: -16.0; beta-H pyrrole: -27.5, -34.3, -41.8 ppm, at 205 K) are without a parallel to any iron(II) porphyrin or heteroporphyrin and indicate a profound alteration of the electronic structure of iron(II) porphyrin upon the coordination of two sigma-aryls. PMID- 15332809 TI - Synthesis, characterization, and X-ray crystal structures of cyclam derivatives. 5. Copper(II) binding studies of a pyridine-strapped 5,12-dioxocyclam-based macrobicycle. AB - The copper(II) binding properties of the macrobicyclic diamide 1,9,12,18,22 pentaazatricyclo[7.6.6.1(3,7)]docosa-3,5,7(22)-triene-13,19-dione (L1) have been fully investigated by spectroscopic (IR, UV-vis, EPR, MALDI-TOF MS), X-ray diffraction, potentiometric, electrochemical, and spectroelectrochemical methods. This constrained receptor possesses a hemispherical cavity created by cross bridging the 1 and 8 positions of trans-dioxocyclam (1,4,8,11 tetraazacyclotetradecane-5,12-dione, L2) with a 2,6-pyridyl strap. Treatment of L1 with a copper salt in methanol produces a red complex of [Cu(L1H(-1))]+ formula in which the copper atom is embedded in a 13-membered ring and coordinated by both amines as well as the pyridine and one deprotonated amide nitrogen atoms. Infrared spectroscopy provides evidence for protonation of the carbonyl oxygen atom belonging to the copper-bound amide of [Cu(L1H(-1))]+ under strongly acidic conditions. The resulting conversion of the amidate into an iminol group highlights the inert character of the corresponding complexes, which do not dissociate at low pH values. In contrast, both secondary amides of L1 deprotonate in the presence of a weak base, thus affording a blue pentacoordinated [Cu(L1H(-2))] compound where the copper atom sits in the center of the 14-membered dioxocyclam fragment. In aqueous solution, both complexes undergo a pH-driven (pK(a) = 8.73(2)) molecular reorganization, which is reminiscent of a glider motion. The copper(II) cation switches rapidly and reversibly from a four-coordinate flattened tetrahedral arrangement of the donor atoms in the red species to a five-coordinate environment in the blue species, which is intermediate between a square pyramid and a trigonal bipyramid. Conversion of the red to the blue form was also demonstrated to occur upon reduction of [Cu(L1H(-1))]+ by cyclic voltammetry (E(pc) = -0.64 V/SCE in CH(3)CN). PMID- 15332810 TI - Silver-109 NMR spectroscopy of inorganic solids. AB - In this study the (109)Ag NMR spectra of the following solid inorganic silver containing compounds were investigated: AgNO(3), AgNO(2), Ag(2)SO(4), Ag(2)SO(3), AgCO(3), Ag(3)PO(4), AgCl, AgBr, AgI, AgSO(3)CH(3), silver p-toluenesulfonate, NaAg(CN)(2), KAg(CN)(2), K(3)Ag(CN)(4), Me(4)NAgCl(2), silver diethylthiocarbamate, silver lactate, silver acetate, silver citrate, and bis[(N,N(1)-di-tert-butylformamidinato)silver(I)]. The magic angle spinning (MAS) spectra of all compounds were obtained. In some cases, when protons were available, the (1)H to (109)Ag cross-polarization (CP) technique was used to enhance the signal and shorten the experimental relaxation delay. It was possible to obtain slow MAS (or CP/MAS) or nonspinning spectra for 10 samples, allowing the determination of the principal components of the (109)Ag chemical shift (CS) tensors. The isotropic chemical shifts and the CS tensors are discussed in light of the available crystal structures. The need for an accepted standard for referencing (109)Ag chemical shifts and the use of AgSO(3)CH(3) as a CP setup sample are also discussed. PMID- 15332811 TI - Structural study by NMR of an oxorhenium-RGD decapeptide complex for application in radiotherapy. AB - The decapeptide Arg-Gly-Asp-Ser-Cys-Arg-Gly-Asp-Ser-Tyr, which contains two Arg Gly-Asp (RGD) moieties in its sequence, has been successfully labeled with radioactive rhenium (Re-188) yielding a single, stable oxorhenium complex. This complex is being evaluated for possible application in oncology as a target specific radiotherapeutic agent, because its radioactive technetium-99m analogue has already been applied for the scintigraphic detection of malignant melanoma in humans. For structural characterization purposes, the complex of the decapeptide was synthesized at the macroscopic level using nonradioactive rhenium (Re-185/Re 187). NMR and mass spectral analysis of the nonradioactive oxorhenium complex revealed that the decapeptide coordinates to the oxorhenium core through the N(amide) of Asp3, the N(amide) of Ser4, and the N(amide) and S(thiolate) atoms of Cys5 to form a complex of the ReO[N(3)S] type. PMID- 15332812 TI - New Ag(I)-containing coordination polymers generated from multidentate Schiff base ligands. AB - The coordination chemistry of the multidentate Schiff-base ligands 2,5-bis(3 methylpyrazinyl)-3,4-diaza-2,4-hexadiene (L5) and 2,5-bis(pyrazinyl)-3,4-diaza 2,4-hexadiene (L6) with inorganic Ag(I) salts has been investigated. Six new Ag(I)-coordination polymers were prepared by solution reactions and fully characterized by infrared spectroscopy, elemental analysis, thermogravimetric analysis, and single-crystal X-ray diffraction. [Ag(L5)]ClO(4).0.5CH(3)OH (1, orthorhombic, Fdd2; a = 20.0896(11) A, b = 48.224(3) A, c = 7.8432(4) A, Z = 16), [Ag(L5)]PF(6).0.5CH(3)OH (2, orthorhombic, Fdd2; a = 20.7255(11) A, b = 46.166(2) A, c = 8.4332(4) A, Z = 16), [Ag(L5)]SbF(6).0.5CH(3)OH (3, orthorhombic, Fdd2; a = 21.5481(11) A, b = 45.196(2) A, c = 8.7331(4) A, Z = 16), and [Ag(L5)](BF(4)).0.5CH(3)OH (4, orthorhombic, Fdd2; a = 19.8897(11) A, b = 48.358(3) A, c = 7.7491(5) A, Z = 16) were obtained by combination of L5 with AgClO(4).xH(2)O, AgPF(6), AgSbF(6), and AgBF(4), respectively, in a methylene chloride/methanol mixed solvent system. Compounds 1-4 are isostructural and feature noninterpenetrating three-dimensional zeolite-like networks. [Ag(4)(L6)(4)](PF(6))(4).CHCl(3) (5, tetragonal, Pc2; a = 16.1067(3) A, b = 16.1067(3) A, c = 14.4935(5) A, Z = 2) was generated from the reaction of L6 with AgPF(6) in a chloroform/ethanol mixed solvent system. It forms with a unique one dimensional nanometer-tube that can be considered a new polymeric motif based on the [AgN5] coordination sphere. The tubes are square with crystallographic dimensions of 10.3 x 10.0 A. The tubes are further linked together through weak interpolymer C-H...F hydrogen bonding interactions into a novel H-bonded three dimensional network containing square tubes, in which uncoordinated PF(6)(-) counterions and chloroform guest molecules are located. Compound 6 ([Ag(mu C(6)H(6)N(2)O)](SO(3)CF(3)), monoclinic, P2(1)/c; a = 12.3435(6) A, b = 20.3548(10) A, c = 9.0861(5) A, Z = 8) was obtained by combination of AgSO(3)CF(3) and L6 in a methylene chloride/benzene mixed solvent system. In 6, 2 acetylpyrazine, which was generated from the hydrolysis reaction of L6 in the presence of CF(3)SO(3)(-) and a small quantity of water in solvent, chelates the Ag(I) centers through the carbonyl O-donor, and the vicinal pyrazine N-donor, furthermore, uses the para-N atoms to link other Ag(I) centers into one dimensional zigzag chains. The triflate anions link the chains into a three dimensional network by somewhat long Ag.O contacts. PMID- 15332813 TI - Red shifting due to nonplanarity in alkylporphyrins: solid-state polarized UV-vis spectra and ZINDO calculations of two nickel(II)octaethylporphyrins. AB - We present the first demonstration of red shifting upon nonplanarity in alkylporphyrins using two pure conformations having known structures with identical substituents. The traditional view about the relationship of spectral red shifting to nonplanar deformation in porphyrins has been that the deformation from planar to nonplanar forms is in itself the cause of the shifting, but recently this view has been challenged. Among the new arguments is that the substituents required to effect conformational change also bring about nuclear rearrangements in the porphyrin complex which is the actual cause of the red shifting. Octaethylporphyrinatonickel(II), however, exists in both planar and ruffled forms which are determined only by the crystal structure, thus making the issue of different substituents moot. Using a polarized specular reflectance UV vis microspectrophotometer, we have obtained polarized spectra of pure, solid samples of both forms of NiOEP. We find Soret band red shifting in the solid state that is much larger than previous reports of solution spectra and also report Q-band red shifting. We performed ZINDO calculations on monomers and dimers of both forms of NiOEP, based upon reported structures, and have reproduced the reported solution transition energies and our solid-state spectra as well as the red shifts that we and others have found experimentally. We conclude that, at least in this system, red shifting does indeed result primarily from conformation changes in the porphyrin. PMID- 15332814 TI - Syntheses, crystal structures, and luminescent properties of novel layered lanthanide sulfonate-phosphonates. AB - Hydrothermal reactions of lanthanide metal salts with MeN(CH(2)CO(2)H)(CH(2)PO(3)H(2)) (H(3)L) and 5-sulfoisophthalic acid monosodium salt (NaH(2)BTS) lead to four isomorphous lanthanide carboxylate-phosphonate sulfonate hybrids, namely, Ln(H(2)L)(HBTS)(H(2)O)(2).H(2)O (Ln = La (1), Pr (2), Nd (3), Gd (4)). Their structures have been established by X-ray single-crystal diffraction. The interconnection of the lanthanide(III) ions by carboxylate phosphonate ligands results in a 1D double chain; these double chains are further bridged by bidentate bridging carboxylate-sulfonate ligands to form a <011> layer. The luminescent properties of compounds 3 and 4 have also been studied. PMID- 15332815 TI - Electrocatalytic proton reduction by phosphido-bridged diiron carbonyl compounds: distant relations to the H-cluster? AB - Intermediates formed during reduction of Fe(2)(mu-PPh(2))(2)(CO)(6) (1) in the presence of protons have been identified by spectroelectrochemical, continuous flow, and interrupted-flow techniques. The mechanism for electrocatalytic proton reduction suggested by these observations yields digital simulation of the voltammetry in close agreement with measurements conducted in THF over a range of acid concentrations. The mechanism for electrocatalytic proton reduction involves initial formation of the dianion, 1(2-), which is doubly protonated prior to further reduction and dihydrogen elimination. The IR spectra of the singly and doubly protonated forms of 1(2-) indicate structures corresponding to [FeH(CO)(3)(mu-PPh(2))(2)Fe(CO)(3)](-) (1H-) and FeH(CO)(3)(mu PPh(2))(2)FeH(CO)(3) (1H(2)). The thiolato and dithiolato analogues of 1 exhibit electrocatalytic proton reduction associated with the two-electron reduction step, and this implies that the corresponding two-electron reduced doubly protonated species is unstable with respect to dihydrogen elimination. The stability of 1H(2) is most likely to be due to the weak interactions between the iron centers of the flattened [2Fe2P] core. Whereas 1H(2) is stable in the absence of a reducing potential, 1H- rearranges rapidly to a product previously described as [Fe(2)(mu-PPh(2))(mu-CO)(PHPh(2))(CO)(5)](-) (1H-(W)). Another protonation product of 1(2-), previously formulated as [Fe(2)(mu-PPh(2))(2)(mu CO)H(CO)(5)](-), has been reformulated as [Fe(2)(mu-PPh(2))(mu-CO)(CO)(6)](-) (2) on the basis of a range of spectroscopic measurements. Solution EXAFS measurements of 1, 1(2-), 1H-(W), and 2 are reported, and these yield model independent Fe-Fe distances of 2.61 (1), 3.58 (1(2-)), 2.58 (1H-(W)), and 2.59 A (2). The presence of an Fe-Fe bond for both 1H-(W) and 2 is a key aspect of the proposed structures, and this strongly supports the deductions based on spectroscopic evidence. The fits of the solution EXAFS to different structural models give statistics in agreement with the proposed structures. PMID- 15332816 TI - Pseudooctahedral complexes of vanadium(III): electronic structure investigation by magnetic and electronic spectroscopy. AB - A variety of physical methods has been used to probe the non-Kramers, S = 1, V(III) ion in two types of pseudooctahedral complexes: V(acac)(3), where acac = anion of 2,4-pentanedione, and VX(3)(thf)(3), where thf = tetrahydrofuran and X = Cl and Br. These methods include tunable frequency and high-field electron paramagnetic resonance (HFEPR) spectroscopy (using frequencies of approximately 95-700 GHz and fields up to 25 T) in conjunction with electronic absorption, magnetic circular dichroism (MCD), and variable-temperature variable-field MCD (VTVH-MCD) spectroscopies. Variable-temperature magnetic susceptibility and field dependent magnetization measurements were also performed. All measurements were conducted on complexes in the solid state (powder or mull samples). The field versus sub-THz wave quantum energy dependence of observed HFEPR resonances yielded the following spin Hamiltonian parameters for V(acac)(3): D = +7.470(1) cm(-1); E = +1.916(1) cm(-1); g(x) = 1.833(4); g(y) = 1.72(2); g(z) = 2.03(2). For VCl(3)(thf)(3), HFEPR detected a single zero-field transition at 15.8 cm(-1) (474 GHz), which was insufficient to determine the complete set of spin Hamiltonian parameters. For VBr(3)(thf)(3), however, a particularly rich data set was obtained using tunable-frequency HFEPR, and analysis of this data set gave the folowing: D = -16.162(6) cm(-1); E = -3.694(4) cm(-1); g(x) = 1.86(1); g(y) = 1.90(1); g(z) = 1.710(4). Analysis of the VTVH-MCD data gave spin Hamiltonian parameters in good agreement with those determined by HFEPR for both V(acac)(3) and VBr(3)(thf)(3) and in rough agreement with the estimate for VCl(3)(thf)(3) (D approximately 10 cm(-1), |E/D| approximately 0.18), together with the finding that the value of D is negative for both thf complexes. The electronic structures of these V(III) complexes are discussed in terms of their molecular structures and the electronic transitions observed by electronic absorption and MCD spectroscopies. PMID- 15332817 TI - Heterometallic lanthanide group 12 metal iodides. AB - Neodymium tri-iodide reacts with Group 12 metal (M; M = Zn, Cd, Hg) iodides to form heterometallic compounds. These Lewis acidic M cleave Nd-I bonds to give either ionic ([(THF)(5)NdI(2)][MI(3)THF]; M = Zn, Cd) or charge-neutral [(THF)(5)NdI(micro(2)I)HgI(3)] compounds. Differences in structure are interpreted primarily in terms of M-L bond strengths, rather than Nd-L bond strengths. Experiments with Yb indicate that if there is any excess iodide present in these syntheses then the most readily isolated product is a triiodide salt, i.e., [(THF)(5)YbI(2)][I(3)]. In conventional solvents the presence of Lewis acid is not required for iodide displacement-from pyridine, "YbI(3)" crystallizes as [(py)(5)YbI(2)][I]. These compounds are potentially useful as heterometallic sources of lanthanide-doped iodide matrixes, they illustrate the ease with which iodides are displaced from lanthanide coordination spheres, and they underscore the complexity associated with using lanthanide iodides as Lewis acid catalysts. PMID- 15332818 TI - Synthesis and reactivity of fluoro complexes: Part 2. Rhodium(I) fluoro complexes with alkene and phosphine ligands. Synthesis of the first isolated rhodium(I) bifluoride complexes. Structure of [Rh3(mu3-OH)2(COD)(3)](HF2) by X-ray powder diffraction. AB - The reaction between [Rh(mu-OH)(COD)](2) (COD = 1,5-cyclooctadiene) and 73% HF in THF gives [Rh(3)(mu(3)-OH)(2)(COD)(3)](HF(2)) (1). Its crystal structure, determined by ab initio X-ray powder diffraction methods (from conventional laboratory data), contains complex trimetallic cations linked together in 1D chains by a mu(3)-OH...F-H-F...HO-mu(3) sequence of strong hydrogen bonds. The complex [Rh(mu-F)(COE)(2)](2) (COE = cyclooctene; 2), prepared by reacting [Rh(mu OH)(COE)(2)](2) with NEt(3).3HF (3:2), has been characterized. Complex 1 reacts with PR(3) (1:3) to give [RhF(COD)(PR(3))] [R = Ph (3), C(6)H(4)OMe-4 (4), (i)Pr (5), Cy (6)] that can be prepared directly by reacting [Rh(mu-OH)(COD)](2) with 73% HF and PR(3) (1:2:2). The reactions of 1 with PPh(3) or Et(3)P have been studied by NMR spectroscopy at different molar ratios. Complexes [RhF(PEt(3))(3)] (7), [RhF(COD)(PEt(3))] (8), and [RhF(PPh(3))(3)] (9) have been detected. The complex [Rh(F)(NBD)(iPr(3)P)] (NBD = norbornadiene; 10) was prepared by the sequential treatment of [Rh(mu-OMe)(NBD)](2) with 1 equiv of NEt(3).3HF and (i)Pr(3)P. The first isolated bifluoride rhodium(I) complexes [Rh(FHF)(COD)(PR(3))] [R = Ph (11), (i)Pr (12), Cy (13)], obtained by reacting fluoro complexes 3, 5, and 6 with NEt(3).3HF (3:1), have been characterized. The crystal structures of 3 and 11 have been determined. PMID- 15332819 TI - Hydrothermal synthesis, crystal structure, conductivity, and thermal decomposition of [Cu(4,4'-bipy)(H2O)(Mo3O10)].H2O. AB - The hydrothermal reaction of (NH(4))(6)Mo(7)O(24).4H(2)O, CuCl(2).2H(2)O, and 4,4'-bipyridine yields bipyridine-ligated copper-trimolybdate monohydrate [Cu(4,4'-bipy)(H(2)O)(Mo(3)O(10))].H(2)O in the monoclinic system with space group of C(2/c) and cell parameters of a = 15.335(2) A, b = 15.535(2) A, c = 15.106(2) A, beta = 101.162(2) degrees, V = 3530.7(9) A(3), and Z = 8. Its structure consists of one-dimensional infinite ([Mo3O10]2-)( infinity ) chains linked through [Cu2(H2O)2(4,4'-bipy)] units. The Mo-O chain contains distorted [MoO(6)] octahedra connected through corner-sharing oxygen atoms into infinite chains along the c direction and each chain is located in the channel formed by four adjacent crossing chains of [Cu(4,4'-bipy)(H2O)](n)(2n+). The crystal shows weak conductivity through Mo-O chain along the c direction and insulating property along either a or b direction. Furthermore, a crystalline bimetallic oxide, CuMo3O10, forms when the title compound undergoes thermal treatment in N(2) atmosphere after the complete removal of the ligands. PMID- 15332820 TI - Structural and functional models of the active site of zinc phosphotriesterase. AB - In an attempt to prepare structural and functional models for the active site of the hydrolytic enzyme zinc phosphotriesterase, five new zinc complexes of the ligands 2,6-bis[N-(N-(carboxylmethyl)-N-((1-methylimidazol)methyl)amine)methyl]-4 methylphenolate (BCIMP) and the corresponding asymmetric ligand 2-(N-isopropyl-N ((1-methylimidazolyl)methyl)aminomethyl)-6-(N-carboxylmethyl-N-((1 methylimidazolyl)methyl)aminomethyl)-4-methylphenol (ICIMP) have been synthesized, viz. Na[Zn(2)(BCIMP)Ac(2)] (1), [Zn(2)(BCIMP)(Ph(2)Ac)] (2), [Zn(2)(ICIMP)Ac(2)] (3), [Zn(4)(ICIMP)(2)(Me(3)Ac)(2)][ClO(4)](2) (4), and [Zn(4)(ICIMP)(2)(Ph(2)Ac)(2)][ClO(4)](2) (5). The X-ray structure of complex 5 has been determined and reveals that the complex is a dimer of dimers in the solid state, which in solution dissociates to potent structural models. Studies using NMR show that only one carboxylate coligand bridges the dizinc units in the case of diphenyl acetate and pivalate, while the steric bulk of acetate is sufficiently small to permit the coordination of two acetates/dizinc unit. Functional studies involving the hydrolysis/transesterification of 2 hydroxypropyl p-nitrophenyl phosphate (HPNP) show that the complex with ICIMP (compound 5) has a significantly higher rate of catalysis than the BCIMP complex (compound 2). This is attributed to the vacant/labile coordination site that is available in the ICIMP complex but not the BCIMP complex. PMID- 15332821 TI - Novel nucleophilic reactivity of disulfido ligands coordinated parallel to M-M (M = Rh, Ir) bonds. AB - Reaction of trans-[(MCp)(2)(mu-CH(2))(2)Cl(2)] (M = Rh, Ir; Cp = eta(5) C(5)Me(5)) with Li(2)S(2) afforded the disulfido complexes [(MCp)(2)(mu CH(2))(2)(mu-S(2)-S:S')] which were easily oxidized by O(2) to give the oxygenated complexes [(MCp)(2)(mu-CH(2))(2)(mu-SSO(2)-S:S')]. Although [(RhCp)(2)(mu-CH(2))(2)(mu-S(2)-S:S')] gave a complicated mixture when reacted with CH(2)Cl(2) or CHCl(3), [(IrCp)(2)(mu-CH(2))(2)(mu-S(2)-S:S')] reacted with both CH(2)Cl(2) and CHCl(3) to give the dithioformato complex [(IrCp)(2)(mu CH(2))(2)(mu-S(2)CH-S:S')]Cl and the cyclotetrasulfido complex [((IrCp)(2)(mu CH(2))(2))(2)(mu-S(4)-S:S':S":S"')]Cl(2). The oxygenated complexes [(RhCp)(2)(mu CH(2))(2)(mu-SSO(2)-S:S')] reacted with hydrocarbyl halides to afford bridging hydrocarbyl thiolato complexes accompanied by the generation of SO(2) gas. These complexes have been characterized by NMR spectroscopy, ESI-MS, and X-ray diffraction. PMID- 15332822 TI - Vibrational study of some layered structures based on titanium and zirconium phosphates. AB - A Raman and infrared study was carried out on layered zirconium and titanium acid phosphates of alpha- and gamma-type, alpha-M[O(3)POH](2).H(2)O and gamma M[PO(4)][O(2)P(OH)(2)].2H(2)O, respectively. The spectra were initially approached by means of the classical correlation method in the solid state, which accounts for the complexity of the infrared spectra of both species. However, the number of bands and their relative intensity in the Raman spectra suggest a quite total absence of quadrupolar coupling between the vibrating units. So, if interunit coupling is neglected, a molecular approach considering the vibrations of isolated tetrahedral [PO(4)] and octahedral [MO(6)] building blocks can allow an affordable spectroscopic description of the title compounds. Interesting insights on the relationships between spectral properties and structure can be drawn by comparison with the spectra of alkali phosphates and of MO(6) oxoanions. A significant high-energy shift of the nu(P-O) modes is observed in the layered phosphates with respect to the corresponding salts, which parallels the low energy shift of the nu(M-O) modes. Surprisingly, an increase of the M-OP interaction can reinforce the P-O bond. A simple theoretical model, based on the interaction between the [PO(4)] unit and four Li(+) in similar geometrical arrangement found in the structures of the layered phosphates, offers a reasonable explanation of this phenomenon. PMID- 15332823 TI - Synthesis, structure, and catalytic properties of V(IV), Mn(III), Mo(VI), and U(VI) complexes containing bidentate (N, O) oxazine and oxazoline ligands. AB - Synthesis of seven complexes containing oxazoline ([(L(1))(2)V=O] (4), [(L(1))(2)MoO(2)] (5), [(L(1))(2)UO(2)] (6); HL(1) (1) [HL(1) = 2-(4',4'-dimethyl 3'-4'-dihydroxazol-2'-yl)phenol]), chiral oxazoline ([(L(2))(2)UO(2)] (7); HL(2) (2) [HL(2) = (4'R)-2-(4'-ethyl-3'4'-dihyroxazol-2'-yl)phenol]), and oxazine ([(L(3))(2)V=O] (8), [(L(3))(2)Mn(CH(3)COO(-))] (9), [(L(3))(2)Co] (10); HL(3) (3) [HL(3) = 2-(5,6-dihydro-4H-1,3-oxazolinyl)phenol]) and their characterization by various techniques such as UV-vis, IR, and EPR spectroscopy, mass spectrometry, cyclic voltammetry, and elemental analysis are reported. The novel oxazine (3) and complexes 4, 5, 8 and 9 were also characterized by X-ray crystallography. Oxazine 3 crystallizes in the monoclinic system with the P2(1)/n space group, complexes 4 and 9 crystallize in the monoclinic system with the P2(1)/c space group, and complexes 5 and 8 crystallize in the orthorhombic system with the C222(1) space group and the P2(1)2(1)2(1) chiral space group, respectively. The representative synthetic procedure involves the reaction of metal acetate or acetylacetonate derivatives with corresponding ligand in ethanol. Addition of Mn(OAc)(2).4H(2)O to an ethanol solution of 3 gave the unexpected complex Mn(L(3))(2).(CH(3)COO(-)) (9) where the acetate group is coordinated with the metal center in a bidentate fashion. The catalytic activity of complexes 4-9 for oxidation of styrene with tert-butyl hydroperoxide was tested. In all cases, benzaldehyde formed exclusively as the oxidation product. PMID- 15332824 TI - Pyridine substituted N-heterocyclic carbene ligands as supports for Au(I)-Ag(I) interactions: formation of a chiral coordination polymer. AB - Reaction of 1,3-bis(2-pyridinylmethyl)-1H-imidazolium tetrafluoroborate, [H(pyCH(2))(2)im]BF(4), with silver oxide in dichloromethane readily yields [Ag((pyCH(2))(2)im)(2)]BF(4), 1.BF(4)(). 1.BF(4) is converted to the analogous Au(I)-containing species, [Au((pyCH(2))(2)im)(2)]BF(4), 3, by a simple carbene transfer reaction in dichloromethane. Further treatment with two equivalents of AgBF(4) produces the trimetallic species [AuAg(2)((pyCH(2))(2)im)(2)(NCCH(3))(2)](BF(4))(3), 4, which contains two silver ions each coordinated to the pyridine moieties on one carbene ligand and to an acetonitrile molecule in a T-shaped fashion. Monometallic [Ag((py)(2)im)(2)]BF(4), 5, and [Au((py)(2)im)(2)]BF(4), 6, are made analogously to 1.BF(4) and 3 starting from 1,3-bis(2-pyridyl)-imidazol-2-ylidene tetrafluoroborate, [H(py)(2)im]BF(4). Addition of excess AgBF(4) to 6 yields the helical mixed-metal polymer, ([AuAg((py)(2)im)(2)(NCCH(3))](BF(4))(2))(n), 7 which contains an extended Au(I)-Ag(I) chain with short metal-metal separations of 2.8359(4) and 2.9042(4) A. Colorless, monometallic [Hg((pyCH(2))(2)im)(2)](BF(4))(2), 8, is easily produced by refluxing [H(pyCH(2))(2)im)]BF(4) with Hg(OAc)(2) in acetonitrile. The related quinolyl substituted imidazole, [H(quinCH(2))(2)im]PF(6), is produced analogously to [H(pyCH(2))(2)im]BF(4). [Hg((quinCH(2))(2)im)(2)](PF(6))(2), 9, is isolated in good yield as a white solid from the reaction of Hg(OAc)(2) and [H(quinCH(2))(2)im]PF(6). The reaction of [H(quinCH(2))(2)im]PF(6) with excess Ag(2)O produces the triangulo-cluster [Ag(3)((quinCH(2))(2)im)(3)](PF(6))(3), 11. All of these complexes were studied by (1)H NMR spectroscopy, and complexes 3-9 were additionally characterized by X-ray crystallography. These complexes are photoluminescent in the solid state and in solution with spectra that closely resemble those of the ligand precursor. PMID- 15332825 TI - Steric and hydrogen-bonding effects on the stability of copper complexes with small molecules. AB - A series of the copper(II) complexes with tripodal tetradentate tris(pyridyl 2 methyl)amine-based ligands possessing the hydrogen-bonding 6-aminopyridine units (tapa, three amino groups; bapa, two amino groups; mapa, one amino group) have been synthesized, and their copper(II) complexes with a small molecule such as dioxygen and azide have been studied spectroscopically and structurally. The reaction of their Cu(II) complexes with NaN(3) have given the mononuclear copper complexes with azide in an end-on mode, [Cu(tapa)(N(3))]ClO(4) (1a), [Cu(bapa)(N(3))]ClO(4) (2a), [Cu(mapa)(N(3))]ClO(4) (3a), and [Cu(tpa)(N(3))]ClO(4) (4a) (tpa, no amino group). The crystal structures have revealed that the coordination geometries around the metal centers are almost a trigonal-bipyramidal rather than a square-planar except for 1a with an intermediate between them. The UV-vis and ESR spectral data indicate that the increase of NH(2) groups of ligands causes the structural change from trigonal bipyramidal to square-pyramidal geometry, which is regulated by a combination of steric repulsion and hydrogen bond. The steric repulsion of amino groups with the azide nitrogen gives rise to elongation of the Cu-N(py) bonds, which leads to the positive shift of the redox potentials of the complexes. The hydrogen bonds between the coordinated azide and amino nitrogens (2.84-3.05 A) contribute clearly to the fixation of azide. The Cu(I) complexes with bapa and mapa ligands have been obtained as a precipitate, although that with tapa was not isolated. The reactions of the Cu(I) complexes with dioxygen in MeOH at -75 degrees C have given the trans-micro-1,2 peroxo dinuclear Cu(II) complexes formulated as [((tapa)Cu)(2)(O(2))](2+) (1c), [((bapa)Cu)(2)(O(2))](2+) (2c), and [((mapa)Cu)(2)(O(2))](2+) (3c), whose characterizations were confirmed by UV-vis, ESR, and resonance Raman spectroscopies. UV-vis spectra of 1c, 2c, and 3c exhibited intense bands assignable to pi(O(2)(2)(-))-to-d(Cu) charge transfer (CT) transitions at lambda(max)/nm (epsilon/M(-1)cm(-1)) = 449 (4620), 474 (6860), and 500 (9680), respectively. The series of the peroxo adducts generated was ESR silent. The resonance Raman spectra exhibited the enhanced features assignable to two stretching vibrations nu((16)O-(16)O/(18)O-(18)O)/cm(-1) and nu(Cu-(16)O/Cu-(18)O)/cm(-1) at 853/807 (1c), 858/812 (2c), 847/800 (3c), and at 547/522 (2c), 544/518 (3c), respectively. The thermal stability of the peroxo copper species has increased with increase in the number of the hydrogen-bonding interactions between the peroxide and amino groups. PMID- 15332826 TI - Syntheses, structures, and reactivities of (Fe-NO)6 nitrosyls derived from polypyridine-carboxamide ligands: photoactive NO-donors and reagents for S nitrosylation of alkyl thiols. AB - Two new iron nitrosyls derived from two designed pentadentate ligands N,N-bis(2 pyridylmethyl)-amine-N'-(2-pyridylmethyl)acetamide and N,N-bis(2-pyridylmethyl) amine-N'-[1-(2-pyridinyl)ethyl]acetamide (PcPy(3)H and MePcPy(3)H, respectively, where H is the dissociable amide proton) have been structurally characterized. These complexes are similar to a previously reported (Fe-NO)6 complex, [(PaPy(3))Fe(NO)](ClO(4))(2) (1) that releases NO under mild conditions. The present nitrosyls, namely [(PcPy(3))Fe(NO)](ClO(4))(2) (2) and [(MePcPy(3))Fe(NO)](ClO(4))(2) (3), belong to the same (Fe-NO)6 family and exhibit (a) clean (1)H NMR spectra in CD(3)CN indicating S = 0 ground state, (b) almost linear Fe-N-O angles (177.3(5) degrees and 177.6(4) degrees for 2 and 3, respectively), and (c) N-O stretching frequencies (nu(NO)) in the range 1900-1925 cm(-)(1). The binding of NO at the non-heme iron centers of 1-3 is completely reversible and all three nitrosyls rapidly release NO when exposed to light (50 W tungsten bulb). In addition to acting as photoactive NO-donors, these complexes also nitrosylate thiols such as N-acetylpenicillamine, 3-mercaptopropionic acid, and N-acetyl-cysteine-methyl-ester in yields that range from 30 to 90% in the absence of light. The addition of alkyl or aryl thiolate (RS(-)) to the (Fe-NO)6 complexes in the absence of dioxygen results in the reduction of the iron metal center to afford the corresponding (Fe-NO)7 species. PMID- 15332827 TI - Electron transfer reaction of oxo(salen)chromium(V) ion with anilines. AB - The kinetics of oxidation of 16 meta-, ortho-, and para-substituted anilines with nine oxo(salen)chromium(V) ions have been studied by spectrophotometric, ESIMS, and EPR techniques. During the course of the reaction, two new peaks with lambda(max) at 470 and 730 nm appear in the absorption spectrum, and these peaks are due to the formation of emeraldine forms of oligomers of aniline supported by the ESIMS peaks with m/z values 274 and 365 (for the trimer and tetramer of aniline). The rate of the reaction is highly sensitive to the change of substituents in the aryl moiety of aniline and in the salen ligand of chromium(V) complexes. Application of the Hammett equation to analyze kinetic data yields a rho value of -3.8 for the substituent variation in aniline and +2.2 for the substituent variation in the salen ligand of the metal complex. On the basis of the spectral, kinetic, and product analysis studies, a mechanism involving an electron transfer from the nitrogen of aniline to the metal complex in the rate controlling step has been proposed. The Marcus equation has been successfully applied to this system, and the calculated values are compliant with the measured values. PMID- 15332828 TI - Utility of anhydrous neodymium nitrate as a precursor to extended organoneodymium nitrate networks. AB - Hydrated neodymium nitrates can be readily transformed to anhydrous ether solvates which react with cyclopentadienyl reagents to make organometallic nitrate complexes with variable degrees of oligomerization. Heating Nd(NO(3))(3)(H(2)O)(6) in tetrahydrofuran at reflux, removal of solvent, drying at 100 degrees C under high vacuum, and addition of hot THF generates Nd(NO(3))(3)(THF)(3), 1. Using dimethoxyethane, Nd(NO(3))(3)(DME)(2), 2, can be obtained similarly. Addition of NaC(5)Me(5) to 1 generates (C(5)Me(5))Nd(NO(3))(3)(THF)Na(THF)(x)complexes which crystallize as ([(C(5)Me(5))(NO(3))(2)Nd(THF)(micro-NO(3))](2)Na(THF)(4))[Na(THF)(6)], 3, or [(C(5)Me(5))Nd(THF)(mu-NO(3))(3)Na(THF)(2)](n), 4, depending on the conditions. The trimetallic Nd(2)Na unit in 3 forms an extended system in 4 via bridging nitrates. Addition of KC(5)Me(5) and 18-crown-6 to 1 generates another extended complex [(C(5)Me(5))Nd(THF)(NO(3))(mu-NO(3))(2)K(18-crown-6)](n), 5, in which an 18-crown-6 ligated potassium links neodymium centers via two bridging nitrates and an agostic interaction between a C(5)Me(5) methyl group and potassium. PMID- 15332829 TI - One-dimensional oxalato-bridged Cu(II), Co(II), and Zn(II) complexes with purine and adenine as terminal ligands. AB - The reaction of nucleobases (adenine or purine) with a metallic salt in the presence of potassium oxalate in an aqueous solution yields one-dimensional complexes of formulas [M(mu-ox)(H(2)O)(pur)](n) (pur = purine, ox = oxalato ligand (2-); M = Cu(II) [1], Co(II) [2], and Zn(II) [3]), [Co(mu ox)(H(2)O)(pur)(0.76)(ade)(0.24)](n)(4) and ([M(mu ox)(H(2)O)(ade)].2(ade).(H(2)O))(n) (ade = adenine; M = Co(II) [5] and Zn(II) [6]). Their X-ray single-crystal structures, variable-temperature magnetic measurements, thermal behavior, and FT-IR spectroscopy are reported. The complexes 1-4 crystallize in the monoclinic space group P2(1)/a (No. 14) with similar crystallographic parameters. The compounds 5 and 6 are also isomorphous but crystallize in the triclinic space group P (No. 2). All compounds contain one dimensional chains in which cis-[M(H(2)O)(L)](2+) units are bridged by bis bidentate oxalato ligands with M(.)M intrachain distances in the range 5.23-5.57 A. In all cases, the metal atoms are six-coordinated by four oxalato oxygen atoms, one water molecule, and one nitrogen atom from a terminal nucleobase, building distorted octahedral MO(4)O(w)N surroundings. The purine ligand is bound to the metal atom through the most basic imidazole N9 atom in 1-4, whereas in 5 and 6 the minor groove site N3 of the adenine nucleobase is the donor atom. The crystal packing of compounds 5 and 6 shows the presence of uncoordinated adenine and water crystallization molecules. The cohesiveness of the supramolecular 3D structure of the compounds is achieved by means of an extensive network of noncovalent interactions (hydrogen bonds and pi-pi stacking interactions). Variable-temperature magnetic susceptibility measurements of the Cu(II) and Co(II) complexes in the range 2-300 K show the occurrence of antiferromagnetic intrachain interactions. PMID- 15332830 TI - Preparation, characterization, and magnetic behavior of the ln derivatives (ln = nd, la) of a 2,6-diiminepyridine ligand and corresponding dianion. AB - An unprecedented Nd[2,6-[[2,6-(i Pr)(2)C(6)H(5)]N=C(CH(3))](2)(C(5)H(3)N)]NdI(2)(THF) (1) complex was prepared by oxidizing metallic Nd with I(2) in THF and in the presence of 2,6-[[2,6-(i Pr)(2)C(6)H(5)]N=C(CH(3))](2)(C(5)H(3)N). The magnetic behavior at variable T clearly indicated that the complex should be regarded as a trivalent Nd atom antiferromagnetically coupled to a radical anion. By using the doubly deprotonated form of the diimino pyridine ligand [[2,6-[[2,6-(i-Pr)(2)C(6)H(5)]N C=CH(2)](2)(C(5)H(3)N)](2-) (2) the corresponding trivalent complexes [[2,6-[[2,6 (i-Pr)(2)C(6)H(5)]N-C=CH(2)](2)(C(5)H(3)N)]Ln (THF)](mu-Cl)(2)[Li(THF)(2)].0.5 (hexane) [Ln = Nd (3), La (4)] were obtained and characterized. Reduction of these species afforded electron transfer to the ligand system which gave ligand dimerization via C-C bond formation through one of the two ene-amido functions of each molecule. The resulting dinuclear [[([2,6-(i-Pr)(2)C(6)H(5)]N C=(CH(2)))(C(5)H(3)N)([2,6-(i-Pr)(2)C(6)H(5)]N=CCH(2))]Ln(THF)(2)(mu Cl)[Li(THF)(3)])(2).2(THF) [Ln = Nd (5), La (6)] were isolated and characterized. PMID- 15332831 TI - New strontium polysulfides, SrS3, and Sr2(OH)2S4.10H2O, obtained by the high pressure treatment of a Sr-S mixture. AB - A new polymorph of SrS(3) was obtained by a reaction of SrS and S with an atomic ratio of Sr:S = 1:5 under a pressure of 5 GPa at 1200 degrees C. It crystallized in a tetragonal unit cell with a = 6.708(1) A, c = 3.942(1) A, and V = 177.36(6) A(3). It was isotypic with BaS(3), and contained S3(2-) polysulfide ions. The product obtained from the high-pressure synthesis contained an amorphous component. It was highly deliquescent and formed a yellowish solution. A new layered polysulfide, Sr(2)(OH)(2)S(4).10H(2)O, crystallized in the solution. The sulfide belonged to a triclinic space group of P (No. 2) with lattice constants of a = 5.9107(5) A, b = 7.8682(6) A, c = 9.4134(6) A, alpha = 75.639(6) degrees, beta = 73.824(3) degrees, gamma = 71.639(3) degrees, V = 392.83(5) A(3), and Z = 1. Each Sr ion was coordinated with one OH ligand and eight H(2)O ligands. Six H(2)O ligands out of the eight were bridging ligands to form two-dimensional [Sr(2)(OH)(2)(H(2)O)(10)(2+)]( infinity ) cationic layers, between which S4(2-) tetrapolysulfide ions were situated. The S4(2-) anion had a coplanar configuration with a dihedral angle of 180.0 degrees. The stability of S4(2-) anions having different conformations was discussed from a viewpoint of ab initio MO calculations on changing the dihedral angles of S4(2-). PMID- 15332833 TI - An analysis of cytotoxic botanical formulations used in the traditional medicine of ancient Persia as abortifacients. AB - We report here an analysis of plants used by traditional healers of ancient Persia to induce abortions. Six herbal formulations that contain 39 different species from 21 plant families with their vernacular names, English names, amounts used, and their methods of preparation are reported. Our initial objective of this ethnobotanical analysis was to evaluate the validity and the efficacy of the plants used by (1) comparing other reported uses of these plants in traditional medicine, (2) investigating the medical and pharmacological literature on the medicinal properties of the plant species used, and (3) investigating the reported cytotoxic effects of compounds prevalent in these plants. Previous phytochemical analyses have shown that a number of plant species are rich in coumarin compounds that have potential antineoplastic or cytotoxic activities. Our results indicate a correlation between the reported use of these plants as abortifacients and their cytotoxic (antineoplastic) effects. In addition, we discuss the process in which this ethnobotanical investigation led to the discovery of dicoumarol (a coumarin anticoagulant) as a potential chemotherapeutic agent. PMID- 15332834 TI - Marine toxins and nonmarine toxins: convergence or symbiotic organisms? AB - Bioactive marine natural products occur only rarely in nonmarine sources. The converse also is true. Divergent evolutionary pathways for the biosynthesis of bioactive secondary metabolites seem to be the rule. Marine biosynthetic pathways lead to a wide variety of different structural classes, among which polyethers, macrolides, terpenes, unusual amino acids/peptides, and alkaloids are notable. Nonmarine biosynthetic pathways also lead to a similar wide variety of structural classes. However, the structures are usually quite different from the marine analogues. The alkaloids of plants are notable, but again there appears little convergence between the marine and nonmarine alkaloids. However, tetrodotoxin, a remarkable, highly polar, marine alkaloid, does occur in various amphibians. The occurrence and possible origin of tetrodotoxin and congeners, including chiriquitoxin, and of the saxitoxin analogue zetekitoxin in amphibians are reviewed. PMID- 15332835 TI - Marine natural products and related compounds in clinical and advanced preclinical trials. AB - The marine environment has proven to be a very rich source of extremely potent compounds that have demonstrated significant activities in antitumor, antiinflammatory, analgesia, immunomodulation, allergy, and anti-viral assays. Although the case can and has been made that the nucleosides such as Ara-A and Ara-C are derived from knowledge gained from investigations of bioactive marine nucleosides, no drug directly from marine sources (whether isolated or by total synthesis) has yet made it to the commercial sector in any disease. However, as shown in this review, there are now significant numbers of very interesting molecules that have come from marine sources, or have been synthesized as a result of knowledge gained from a prototypical compound, that are either in or approaching Phase II/III clinical trials in cancer, analgesia, allergy, and cognitive diseases. A substantial number of other potential agents are following in their wake in preclinical trials in these and in other diseases. PMID- 15332836 TI - Research achievement award--D. John Faulkner. AB - David John Faulkner, one of the pioneers of marine natural products chemistry and the 2003 recipient of the ASP Research Achievement Award, passed away on November 23, 2002. John was very pleased to learn that he'd been named as the award recipient and was intending to present the ASP Research Achievement Award Address at the annual meeting in July 2003, Chapel Hill, North Carolina. John's untimely death left an unprecedented event in the history of the ASP-a posthumous Research Achievement Award to a deserving individual and an untimely loss for us all. We are bereft of a colleague, a friend, and a mentor, and the opportunity to hear John's words on the occasion of his own award. In tribute to John, we have assembled a retrospective of John's work that is not meant to be a comprehensive review (this would take considerably more space) but a selection of highlights and personal vignettes from some of those that trained in marine natural products under his mentorship. This paper is a written account of the symposium presented by the authors at the ASP Annual Meeting in Chapel Hill, North Carolina, on July 16, 2003. PMID- 15332837 TI - Antineoplastic Agents. 510. Isolation and structure of dolastatin 19 from the Gulf of California sea hare Dolabella auricularia. AB - The Gulf of California shell-less mollusc Dolabella auricularia has been found to contain a new 14-membered macrocyclic lactone linked to a 2,4-di-O-methyl-l-alpha rhamnopyranoside, designated dolastatin 19 (1). The new cancer cell growth inhibitor (1, 8.33 x 10(-8)% yield) was obtained by bioassay (P388 lymphocytic leukemia and human cancer cell lines) directed isolation, accompanied by debromoaplysiatoxin (9.17 x 10(-7)% yield) and anhydrodebromoaplysiatoxin (2.0 x 10(-7)% yield). The structures were determined on the basis of analyses of high resolution mass spectra and high-field NMR data. All the relative stereochemistry for the chiral centers was designated by utilizing NMR techniques. PMID- 15332838 TI - An analysis of phakellin and oroidin structures stimulated by further study of an Agelas sponge. AB - Two new phakellin alkaloids, (-)-7-N-methyldibromophakellin (14) and (-)-7-N methylmonobromophakellin (15), were isolated from an Agelas sp. sponge, collected near Wewak, Papua New Guinea. Inhibition assays employing both 12- and 15-human lipoxygenase isozymes (12-HLO, 15-HLO) were used to guide the isolation of 14, and LCMS data pointed the way to uncovering 15. The structure elucidations were completed by spectroscopic data analysis and comparisons to the properties of known phakellins. The lipoxygenase IC50 data showed that 14 was modest in its selective inhibition of 12-HLO. The phakellin family is uniquely marine-derived, and comments are offered on the biogenetic insights provided by these new structures. PMID- 15332839 TI - Nagelamides A-H, new dimeric bromopyrrole alkaloids from marine sponge Agelas species. AB - Eight new dimeric bromopyrrole alkaloids, nagelamides A-H (1-8) and a monomeric one, 9,10-dihydrokeramadine (9), have been isolated from the Okinawan marine sponge Agelas sp., and the structures were elucidated from spectroscopic data. Nagelamides A-H (1-8) exhibited antibacterial activity against Gram-positive bacteria. Nagelamide G (7) inhibited protein phosphatase 2A activity. PMID- 15332840 TI - Cytotoxic pyrroloiminoquinones from four new species of South African latrunculid sponges. AB - An examination of organic extracts of four new species of South African latrunculid sponges, Tsitsikamma pedunculata, T. favus, Latrunculia bellae, and Strongylodesma algoaensis, yielded 13 known and eight new pyrroloiminoquinone alkaloids, 3-dihydro-7,8-dehydrodiscorhabdin C (4), 14-bromo-3-dihydro-7,8 dehydrodiscorhabdin C (5), discorhabdin V (6), 14-bromo-1-hydroxydiscorhabdin V (7), tsitsikammamine A N-18 oxime (10), tsitsikammamine B N-18 oxime (11), 1 methoxydiscorhabdin D (12), and 1-aminodiscorhabdin D (13). Standard spectroscopic methods provided the structures of the pyrroloiminoquinone metabolites, while chiral GC-MS analysis of the acylated ozonolysis products of 21 confirmed the stereochemistry of the l-histidine residue in this compound. The anticancer activity of 20 pyrroloiminoquinone compounds was explored in the HCT 116 cancer cell line screen, and the DNA intercalation of the tsitsikammamines, together with their ability to cleave DNA through topoisomerase I inhibition, is discussed. PMID- 15332841 TI - Nematocidal thiocyanatins from a southern Australian marine sponge Oceanapia sp. AB - Investigations of a southern Australian marine sponge, Oceanapia sp., have yielded two new beta methyl branched bisthiocyanates, thiocyanatins D1 (3a) and D(2) (3b), along with two new thiocarbamate thiocyanates, thiocyanatins E1 (4a) and E2 (4b). The new thiocyanatins belong to a rare class of bioactive marine metabolite previously only represented by thiocyanatins A-C (1, 2a/b). Structures were assigned on the basis of detailed spectroscopic analysis, with comparisons to the known bisthiocyanate thiocyanatin A (1) and synthetic model compounds (5 7). The thiocyanatins exhibit potent nematocidal activity, and preliminary structure-activity relationship investigations have confirmed key characteristics of the thiocyanatin pharmacophore. PMID- 15332842 TI - Palmitoylputrescine, an antibiotic isolated from the heterologous expression of DNA extracted from bromeliad tank water. AB - Heterologous expression of large fragments of microbial DNA extracted directly from environmental samples (environmental DNA, or eDNA) in easily cultured hosts should provide access to some of the natural products produced by previously uncultured bacteria. The natural product antibiotic palmitoylputrescine (1) was isolated from Escherichia coli transformed with a cosmid (pCSLF16) containing DNA extracted directly from Costa Rican bromeliad tank water. In this report we describe the characterization of this antibiotic and its biosynthetic gene. PMID- 15332843 TI - Reversible antifouling effect of the cyclotide cycloviolacin O2 against barnacles. AB - Cycloviolacin O2, a plant peptide of the cyclotide family, is shown to have potent effects against fouling barnacles (Balanus improvisus), with complete inhibition of settlement at a concentration of 0.25 microM. The effect of cycloviolacin O2 against barnacles is reversible and nontoxic in the bioassay employed in these studies. Cycloviolacin O2 was isolated from the terrestrial plant Viola odorata by strong cation exchange and reversed-phase HPLC and identified by mass spectrometry following aminoethylation and enzymatic cleavage. PMID- 15332844 TI - Dysinosins B-D, inhibitors of factor VIIa and thrombin from the Australian sponge Lamellodysidea chlorea. AB - Three new marine natural products, dysinosins B-D (1-3), were isolated from the sponge Lamellodysidea chlorea and their structures determined by 1D and 2D NMR spectroscopy. These compounds are inhibitors of the blood coagulation cascade serine proteases factor VIIa and thrombin. These analogues, dysinosins B-D (1-3), allowed identification of two structural motifs within the structures that contribute to binding to the proteases, factor VIIa and thrombin. PMID- 15332845 TI - Chemical investigation of predator-deterred macroalgae from the Antarctic peninsula. AB - Chemical investigation of five Antarctic macroalgae whose tissues and crude extracts displayed ecologically relevant feeding deterrence in field bioassays was performed. Eleven compounds were characterized from the three red algae studied, of which four (1-3 and 9) were previously unreported, and four compounds were found from two brown algae, two (12 and 14) of which are new natural products. Several of these pure compounds have been individually investigated in ecological and/or pharmacological bioassays. PMID- 15332846 TI - Isoprenoids of the soft coral Sarcophyton glaucum: Nyalolide, a new biscembranoid, and other terpenoids. AB - The chemical content of Sarcophyton glaucum, one of the more abundant soft corals on many coral reefs, collected from many seas, was thoroughly explored, resulting in the discovery of a large number of cembranoids, biscembranoids, sterols, and other secondary metabolites. The presently investigated Kenyan specimens of S. glaucum yielded three new metabolites, i.e., nyalolide (15), a biscembranoid, 16 oxosarcoglaucol acetate (16), a cembranoid, and the sesquiterpene guaiacophine (17). Nyalolide was also isolated from the Kenyan soft coral Sarcophyton elegans. The structures of the new compounds were elucidated by interpretation of their MS and 1D and 2D NMR experiments and, in the case of nyalolide, possessing 11 chiral centers, secured by X-ray diffraction analysis. PMID- 15332847 TI - Highly cytotoxic metabolites from the culture supernatant of the temperate dinoflagellate Protoceratium cf. reticulatum. AB - In the course of our search for antitumor compounds in dinoflagellates, the culture broths of two strains of Protoceratium cf. reticulatum showed extremely potent cytotoxicity against human tumor cell lines. The four equally active principles, named protoceratins I (1), II (2), III (3), and IV (4), were purified and their structures were studied. The major principle, protoceratin I (1), proved to be identical with 2-homoyessotoxin, a well-known shellfish toxin. Protoceratins II (2), III (3), and IV (4) were determined to be di-, mono-, and triarabinoside of 1, respectively. They are the first examples of glycosides of dinoflagellate polyethers. PMID- 15332848 TI - Three new manzamine alkaloids from a common Indonesian sponge and their activity against infectious and tropical parasitic diseases. AB - Three new manzamine-type alkaloids, 12,34-oxamanzamine E (3), 8-hydroxymanzamine J (4), and 6-hydroxymanzamine E (8), as well as 12 previously characterized manzamine alkaloids have been isolated from a common Indonesian sponge of the genus Acanthostrongylophora. The structures of the new compounds have been established on the basis of 1D and 2D NMR spectroscopic analysis and comparison of the data to literature values of related compounds. The biological activities and structure-activity relationship of the manzamines against malaria, Mycobacterium tuberculosis, Leishmania, HIV-1, and AIDS opportunistic infections are discussed. A plausible pathway for the formation of the 12,34-oxaether bridge in compound 3 is also provided. PMID- 15332849 TI - Inhibition of HIV-1 envelope-mediated fusion by synthetic batzelladine analogues. AB - Marine natural products that feature polycyclic guanidine motifs, such as crambescidins and batzelladines, are known to have antiviral activities toward some viruses including HSV and HIV. In this study we evaluated a synthetic library containing 28 batzelladine analogues, the structures of which encompass and surpass variations seen in natural batzelladines, for their ability to inhibit HIV-1 envelope-mediated cell-cell fusion. Clear structure-activity relationships were revealed and indicated that the best inhibitors of fusion were most similar in structure to natural batzelladine F, with IC50 values ranging from 0.8 to 3.0 microM. Proceeding from the earlier finding that some batzelladines block gp120-CD4 binding, modeling studies of inhibitors binding to the CD4 binding site on gp120 were carried out. The lowest energy models suggest a preferred orientation for inhibitor binding that is consistent with the observed structure-activity relationships. PMID- 15332850 TI - Synthesis, conformational analysis, and cytotoxicity of new analogues of the natural cyclodepsipeptide jaspamide. AB - Three analogues of the natural bioactive cyclodepsipeptide jaspamide (3-5) were efficiently synthesized using a combination of solid and solution phase techniques. The preliminary design of the molecules has involved the rational substitution and/or simplification of the most critical structural features of the lead compound. The synthetic products were subjected to pharmacological assays, and the conformational properties were investigated by MM (molecular mechanics) and MD (molecular dynamics) calculations, to describe the potential pharmacophoric core responsible for the observed activities. PMID- 15332851 TI - Kulokekahilide-2, a cytotoxic depsipeptide from a cephalaspidean mollusk Philinopsis speciosa. AB - A cytotoxic depsipeptide, kulokekahilide-2 (1), was isolated from a cephalaspidean mollusk, Philinopsis speciosa. The structure elucidation of kulokekahilide-2 was carried out by spectroscopic analysis and chemical degradation. Kulokekahilide-2 showed potent cytotoxicity against several cell lines (P388, SK-OV-3, MDA-MB-435, and A-10 with IC50 values ranging from 4.2 to 59.1 nM) indicating cancer cell selectivity. PMID- 15332852 TI - Genetic evidence supports secondary metabolic diversity in Prochloron spp., the cyanobacterial symbiont of a tropical ascidian. AB - Didemnid family ascidians commonly harbor obligate cyanobacterial symbionts, Prochloron spp., which have been proposed to biosynthesize cyclic peptides. Here, it is shown that Prochloron spp. do indeed contain genes for nonribosomal peptide biosynthesis, although genes for cyclic peptide biosynthesis have not yet been characterized. A peptide synthetase-containing open reading frame of unknown function was cloned from the Prochloron symbionts of some didemnid ascidians, but not from others. These data indicate that Prochloron spp. have variable secondary metabolic potential. PMID- 15332853 TI - Penasulfate A, a new alpha-glucosidase inhibitor from a marine sponge Penares sp. AB - A new alpha-glucosidase inhibitor, penasulfate A, has been isolated from a marine sponge Penares sp.(1) Its structure was elucidated by spectral and chemical methods to be a scalemic mixture of methyl pipecolates acylated with a novel sulfated fatty acid. PMID- 15332854 TI - Lasonolides C-g, five new lasonolide compounds from the sponge Forcepia sp. AB - Five new marine-derived macrolide compounds, lasonolides C (3), D (4), E (5), F (6), and G (7), have been isolated from the sponge Forcepia sp. along with the parent compound in the series, lasonolide A (1). Their structures were elucidated on the basis of spectral data. Compounds 3-5 inhibit the in vitro proliferation of A-549 human lung adenocarcinoma cells with IC50's of 0.13, 4.5, and 0.31 microM, respectively. Compounds 3-6 inhibit the in vitro proliferation of PANC-1 human pancreatic carcinoma cells with IC50's of 0.38, 4.89, 0.57, and 15.6 microM, respectively. Compound 3 inhibits the in vitro proliferation of the NCI ADR-RES cell line with an IC50 of 1.12 microM. PMID- 15332855 TI - Biosynthetic pathway and gene cluster analysis of curacin A, an antitubulin natural product from the tropical marine cyanobacterium Lyngbya majuscula. AB - Curacin A (1) is a potent cancer cell toxin obtained from strains of the tropical marine cyanobacterium Lyngbya majuscula found in Curacao. Its structure is unique in that it contains the sequential positioning of a thiazoline and cyclopropyl ring, and it exerts its potent cell toxicity through interaction with the colchicine drug binding site on microtubules. A series of stable isotope-labeled precursors were fed to cultures of curacin A-producing strains and, following NMR analysis, allowed determination of the metabolic origin of all atoms in the natural product (one cysteine, 10 acetate units, two S-adenosyl methionine derived methyl groups) as well as several unique mechanistic insights. Moreover, these incorporation experiments facilitated an effective gene cloning strategy that allowed identification and sequencing of the approximately 64 kb putative curacin A gene cluster. The metabolic system is comprised of a nonribosomal peptide synthetase (NRPS) and multiple polyketide synthases (PKSs) and shows a very high level of collinearity between genes in the cluster and the predicted biochemical steps required for curacin biosynthesis. Unique features of the cluster include (1) all but one of the PKSs are monomodular multifunctional proteins, (2) a unique gene cassette that contains an HMG-CoA synthase likely responsible for formation of the cyclopropyl ring, and (3) a terminating motif that is predicted to function in both product release and terminal dehydrative decarboxylation. PMID- 15332856 TI - Briarane diterpenes from two species of octocorals, Ellisella sp. and Pteroeides sp. AB - Eight new briarane diterpenes (1-4, 7-10) have been isolated from two species of octocorals and the structures elucidated by spectroscopic analysis. Two diterpenes (2, 3) from the gorgonian Ellisella sp. inhibited cytokinesis, causing multinuclei formation on NBT-II cells, while a known briarane (12) from the sea pen Pteroeides sp. showed reversal of multidrug resistance. PMID- 15332857 TI - Isolation and structure assignments of rostratins A-D, cytotoxic disulfides produced by the marine-derived fungus Exserohilum rostratum. AB - Four new cytotoxic disulfides, rostratins A-D (1-4), were isolated from the whole broth of the marine-derived fungus Exserohilum rostratum (Drechsler), a fungal strain found associated with a marine cyanobacterial mat. The structures of these cyclic dipeptides were established through chemical degradation and a variety of two-dimensional NMR techniques. The absolute configurations of the rostratins were determined by the modified Mosher method. In the case of the polyhydroxylated compound 1 and the mercaptol 4, regioselective acylation was achieved by modulating the reaction temperature while monitoring the progress of the reaction by 1H NMR. Rostratins A, B, C, and D showed in vitro cytotoxicity against human colon carcinoma (HCT-116) with IC50 values of 8.5, 1.9, 0.76, and 16.5 microg/mL, respectively. PMID- 15332858 TI - A concise synthesis of siphonodictidine. AB - Siphonodictidine (1) has been synthesized for the first time in a concise and regiocontrolled manner by using 2-(tert-butyldimethylsiloxy)-3-methylfuran (6) as the crucial building block. The silver trifluoroacetate-induced alkylation of 6 with omega-bromogeranyl acetate 7 gave the key gamma-lactone intermediate 8, which on subsequent reduction, conversion of the hydroxyl into the amino group, and amidination afforded siphonodictidine (1) in an overall yield of 25.7% from 6. PMID- 15332859 TI - Strongylophorine-26, a new meroditerpenoid isolated from the marine sponge Petrosia (Strongylophora) corticata that exhibits anti-invasion activity. AB - Strongylophorine-26 (1), a new meroditerpenoid that shows activity in an anti invasion assay, has been isolated from the marine sponge Petrosia (Strongylophora) corticata collected in Papua New Guinea. The structure of 1 was elucidated by analysis of spectroscopic data. PMID- 15332860 TI - Oxidation of tyrosine diketopiperazine to DOPA diketopiperazine with tyrosine hydroxylase. AB - The diketopiperazine of DOPA was synthesized in high yield from the diketopiperazine of tyrosine using PC12 cell lysate, which expresses high levels of tyrosine hydroxylase. This represents the first use of this enzyme to prepare DOPA-containing peptides. PMID- 15332861 TI - Novel sesquiterpenoid matrix metalloproteinase-3 inhibitors from an acid mine waste extremophile. AB - Berkeley Pit Lake is a 1500 ft deep abandoned open-pit copper mine filled with 30 billion gallons of acidic, metal-contaminated water. This harsh environment is proving to be a source of unusual, biologically active microorganisms. Bioassay guided fractionation using signal transduction enzyme assays led to the isolation of three novel bisabolane sesquiterpenes and a novel coumarin. The isolation and characterization of these compounds are reported here. PMID- 15332862 TI - Dictyosphaeric acids A and B: new decalactones from an undescribed Penicillium sp. obtained from the alga Dictyosphaeria versluyii. AB - Fungal isolate F01V25 was obtained from the alga Dictyosphaeria versluyii collected near Dravuni, Fiji, in 2001 and represented a previously undescribed Penicillium sp. Fermentation of isolate F01V25 resulted in the production of two new polyketides, dictyosphaeric acids A and B, along with the known anthraquinone carviolin. The relative stereochemistry of dictyosphaeric acids A and B was determined using the J-based configuration analysis method in conjunction with ROE and NOE correlations. PMID- 15332863 TI - Aureoverticillactam, a novel 22-atom macrocyclic lactam from the marine actinomycete Streptomyces aureoverticillatus. AB - During the course of our screening program designed to discover novel anticancer and anti-infective agents from marine microorganisms, a strain of Streptomyces aureoverticillatus (NPS001583) isolated from a marine sediment was found to produce a novel macrocyclic lactam with cytotoxicity against various tumor cell lines. Using extensive MS, UV, and NMR spectral analyses, the structure has been established as compound 1, aureoverticillactam, a 22-atom macrocyclic lactam incorporating both triene and tetraene conjugated olefins. PMID- 15332864 TI - Isolation and structure determination of cryptophycins 38, 326, and 327 from the terrestrial cyanobacterium Nostoc sp. GSV 224. AB - Cryptophycin-38 (2), -326 (3), and -327 (4) are three new trace constituents of the terrestrial cyanobacterium Nostoc sp. GSV 224. Cryptophycin-38 is a stereoisomer of cryptophycin-1 (1) and to date is the only naturally occurring analogue that possesses a S,S epoxide group in unit A. Cryptophycin-327 is a geometric isomer that differs from 1 in having a cis Delta(2)-double bond in unit A. Cryptophycin-326 is related to cryptophycin-21, but has two chlorines ortho to the methoxy group in unit B. The relative and absolute stereochemistries of 2 have been related to known cryptophycins by semisynthesis and/or spectral analysis. PMID- 15332865 TI - Neamphamide A, a new HIV-inhibitory depsipeptide from the Papua New Guinea marine sponge Neamphius huxleyi. AB - A new HIV-inhibitory cyclic depsipeptide, neamphamide A (2), was isolated from a Papua New Guinea collection of the marine sponge Neamphius huxleyi. Its structure was established through interpretation of spectroscopic data and by acid hydrolysis, derivatization of the free amino acids, and LC-MS analysis of the derivatives. Neamphamide A (2) contains 11 amino acid residues and an amide linked 3-hydroxy-2,4,6-trimethylheptanoic acid moiety. The amino acid constituents were identified as L-Leu, L-NMeGln, D-Arg, D- and L-Asn, two residues of D-allo-Thr, L-homoproline, (3S,4R)-3,4-dimethyl-L-glutamine, beta methoxytyrosine, and 4-amino-7-guanidino-2,3-dihydroxyheptanoic acid. In a cell based XTT assay, 2 exhibited potent cytoprotective activity against HIV-1 infection with an EC50 of approximately 28 nM. PMID- 15332866 TI - Structure of bryostatin 20: a symbiont-produced chemical defense for larvae of the host bryozoan, Bugula neritina. AB - Larvae of the marine bryozoan Bugula neritina are defended against potential predators by high concentrations of bryostatins, which are produced by a bacterial symbiont of the bryozoan. From the larvae of B. neritina, three bryostatins, bryostatin 10 (1), the novel bryostatin 20 (2), and an as yet uncharacterized bryostatin, were isolated that were unpalatable to fish. These deterrent bryostatins represent the first example from the marine environment of a microbial symbiont producing an antipredator defense for its host. The structure of bryostatin 20 (2) was determined by spectral comparison with previously described bryostatins. PMID- 15332867 TI - Cytotoxic lissoclimide-type diterpenes from the molluscs Pleurobranchusalbiguttatus and Pleurobranchus forskalii. AB - Three new chlorinated diterpenes, 6-8, along with five known ones, 1-5, were isolated from the molluscs Pleurobranchus albiguttatus and P. forskalii collected in the Philippines. These diterpenes are presumably metabolites of a Lissoclinum species of ascidian on which the molluscs have fed. The structures of the new compounds were determined by interpretation of their spectral data. Compounds 1 and 2 were found to be potent cytotoxins in the National Cancer Institute's screening panel of 60 tumor cell lines and showed some selectivity for melanomas. Two other samples exhibited solid tumor selectivity in a soft agar disk diffusion assay. PMID- 15332868 TI - Heterologous biosynthesis of truncated hexaketides derived from the actinorhodin polyketide synthase. AB - Heterologous expression of the actinorhodin polyketide synthase in the recombinant host Streptomyces lividans K4-114 led to the characterization of three new minor polyketides, the novel hexaketides BSM1 and BSM3 and 9' hydroxyaloesaponarin II, in addition to known anthraquinone and aromatic octaketides. The structures of BSM1 and BSM3 imply that these compounds are derived from a C-5-reduced hexaketide intermediate, suggesting that the timing of the ketoreduction reaction in the actinorhodin biosynthetic pathway may take place during the polyketide elongation process rather than after the completion of the octaketide chain as previously suggested. PMID- 15332869 TI - New sphingosines from a gorgonian, Pseudopterogorgia australiensis Ridley, of the Indian Ocean. AB - Two new sphingosines, (2S,3R)-2-(docosanoyl amino)nonadecane-1,3-diol (1) and (2S,3S,4R)-2-[(2'R)-2'-hydroxynonadecanoylamino]nonadecane-1,3,4-triol (3), along with the known (2S,3R,4E)-2-(heptadecanoylamino)octadec-4-ene-1,3-diol, have been isolated from Pseudopterogorgia australiensis, of the Indian Ocean. The structures were deduced from spectral and chemical methods. Compounds 1-3 showed moderate antibacterial activity. PMID- 15332870 TI - Absolute configuration of a hydroxyfuranoid acid from the pelage of the genus Bos, 18-(6S,9R,10R)-bovidic acid. AB - Pelage extracts of the banteng (Bos javanicus), the domestic cattle (B. taurus), the gaur (B. frontalis), and the yak (B. grunniens) were investigated by FABMS and NMR. An 18-carbon hydroxyfuranoid acid, 1 (10-hydroxy-6,9-oxidooctadecanoic acid), first reported from the wool of the domestic sheep (Ovis aries) was confirmed in B. frontalis; we suspend judgment on the occurrence of this compound in the other species we examined. The stereochemistry of 1, determined by Mosher NMR and CD tweezer methods, was assigned as 6S, 9R, 10R. We propose the name bovidic acid for this and homologous alpha-hydroxylated 2,5-tetrahydrofuranoid carboxylic acids, in reference to the family Bovidae, which represents their sole known natural source. PMID- 15332871 TI - Diazepinomicin, a new antimicrobial alkaloid from a marine Micromonospora sp. AB - The structure of a new dibenzodiazepine alkaloid, diazepinomicin (1), isolated from the culture of a marine actinomycete of the genus Micromonospora was characterized using spectroscopic methods. Diazepinomicin represents a unique molecular class composed of a dibenzodiazepine core linked to a farnesyl side chain. PMID- 15332872 TI - Fumagiringillin, a new fumagillin derivative from a strain of the fungus Aspergillus fumigatus. AB - A new fumagillin derivative, fumagiringillin (1), has been isolated together with fumagillin (2) and 12alpha-fumitremorgin C (3) from a strain of Aspergillus fumigatus. The structures were elucidated by spectral analyses. PMID- 15332873 TI - Molecular description of the collapse of hydrophobic polymer chains in water. AB - We propose a self-consistent molecular theory of conformational properties of flexible polymers in solution. It is applied to the collapse of a hydrophobic polymer chain in water, and can be readily generalized to any polymer-solvent system (e.g., copolymers with high complexity). We stress the potential of this method for a variety of problems, such as protein folding. PMID- 15332874 TI - Rate constants from the reaction path Hamiltonian. I. Reactive flux simulations for dynamically correct rates. AB - As ab initio electronic structure calculations become more accurate, inherent sources of error in classical transition state theory such as barrier recrossing and tunneling may become major sources of error in calculating rate constants. This paper introduces a general method for diabatically constructing the transverse eigensystem of a reaction path Hamiltonian in systems with many degenerate transverse frequencies. The diabatically constructed reaction path Hamiltonian yields smoothly varying coupling constants that, in turn, facilitate reactive flux calculations. As an example we compute the dynamically corrected rate constant for the chair to boat interconversion of cyclohexane, a system with 48 degrees of freedom and a number of degenerate frequencies. The transmission coefficients obtained from the reactive flux simulations agree with previous results that have been calculated using an empirical potential. Furthermore, the calculated rate constants agree with experimental values. Comparison to variational transition state theory shows that, despite finding the true bottleneck along the reaction pathway, variational transition state theory only accounts for half of the rate constant reduction due to recrossing trajectories. PMID- 15332875 TI - Rate constants from the reaction path Hamiltonian. II. Nonseparable semiclassical transition state theory. AB - For proton transfer reactions, the tunneling contributions to the rates are often much larger than thermally activated rates at temperatures of interest. A number of separable tunneling corrections have been proposed that capture the dependence of tunneling rates on barrier height and imaginary frequency size. However, the effects of reaction pathway curvature and barrier anharmonicity are more difficult to quantify. The nonseparable semiclassical transition state theory (TST) of Hernandez and Miller [Chem. Phys. Lett. 214, 129 (1993)] accounts for curvature and barrier anharmonicity, but it requires prohibitively expensive cubic and quartic derivatives of the potential energy surface at the transition state. This paper shows how the reaction path Hamiltonian can be used to approximate the cubic and quartic derivatives used in nonseparable semiclassical transition state theory. This enables tunneling corrections that include curvature and barrier anharmonicity effects with just three frequency calculations as required by a conventional harmonic transition state theory calculation. The tunneling correction developed here is nonseparable, but can be expressed as a thermal average to enable efficient Monte Carlo calculations. For the proton exchange reaction NH2 + CH4 <==> NH3 + CH3, the nonseparable rates are very accurate at temperatures from 300 K up to about 1000 K where the TST rate itself begins to diverge from the experimental results. PMID- 15332876 TI - Accurate and highly efficient calculation of the highly excited pure OH stretching resonances of O(1D)HCl, using a combination of methods. AB - Accurate calculation of the energies and widths of the resonances of HOCl--an important intermediate in the O(1D)HCl reactive system--poses a challenging benchmark for computational methods. The need for very large direct product basis sets, combined with an extremely high density of states, results in difficult convergence for iterative methods. A recent calculation of the highly excited OH stretch mode resonances using the filter diagonalization method, for example, required 462,000 basis functions, and 180,000 iterations. In contrast, using a combination of new methods, we are able to compute the same resonance states to higher accuracy with a basis less than half the size, using only a few hundred iterations-although the CPU cost per iteration is substantially greater. Similar performance enhancements are observed for calculations of the high-lying bound states, as reported in a previous paper [J. Theo. Comput. Chem. 2, 583 (2003)]. PMID- 15332877 TI - An explicitly correlated second order Moller-Plesset theory using a frozen Gaussian geminal. AB - A variant of the MP2-R12 class of theories is introduced using an arbitrary geminal function in the place of r12. Integrals are derived for the case where the geminal is expanded in a basis of Gaussian functions in the interelectronic distance. Recurrence relations are derived that do not depend on the exponents of the Gaussian geminals, allowing much of the integration work to be performed after summations over the geminal expansion. Sample calculations at various levels of explicitly correlated MP2 theory are presented for He, Ne, and water. PMID- 15332878 TI - Unbiased expectation values from diffusion quantum Monte Carlo simulations with a fixed number of walkers. AB - We append forward walking to a diffusion Monte Carlo algorithm which maintains a fixed number of walkers. This removes the importance sampling bias of expectation values of operators which do not commute with the Hamiltonian. We demonstrate the effectiveness of this approach by employing three importance sampling functions for the hydrogen atom ground state, two very crude. We estimate moments of the electron-nuclear distance, static polarizabilities, and high-order hyperpolarizabilites up to the fourth power in the electric field, where no use is made of the finite field approximation. The results agree with the analytical values, with a statistical error which increases substantially with decreasing overlap of the guiding function with the exact wave function. PMID- 15332879 TI - Local properties of quantum chemical systems: the LoProp approach. AB - A new method is presented, which makes it possible to partition molecular properties like multipole moments and polarizabilities, into atomic and interatomic contributions. The method requires a subdivision of the atomic basis set into occupied and virtual basis functions for each atom in the molecular system. The localization procedure is organized into a series of orthogonalizations of the original basis set, which will have as a final result a localized orthonormal basis set. The new localization procedure is demonstrated to be stable with various basis sets, and to provide physically meaningful localized properties. Transferability of the methyl properties for the alkane series and of the carbon and hydrogen properties for the benzene, naphtalene, and anthracene series is demonstrated. PMID- 15332880 TI - Reconciling semiclassical and Bohmian mechanics. I. Stationary states. AB - The semiclassical method is characterized by finite forces and smooth, well behaved trajectories, but also by multivalued representational functions that are ill behaved at caustics. In contrast, quantum trajectory methods--based on Bohmian mechanics (quantum hydrodynamics)--are characterized by divergent forces and erratic trajectories near nodes, but also well-behaved, single-valued representational functions. In this paper, we unify these two approaches into a single method that captures the best features of both, and in addition, satisfies the correspondence principle. Stationary eigenstates in one degree of freedom are the primary focus, but more general applications are also anticipated. PMID- 15332881 TI - Mechanism analysis of controlled quantum dynamics in the coordinate representation. AB - There are increasing numbers of successful numerical simulations as well as laboratory tests of controlled quantum phenomena. However, little is understood about the control mechanisms, especially in strongly driven systems. In this paper, mechanism is defined in terms of the significant quantum pathways induced by the control field, connecting the initial state to the final target state. The present work focuses on understanding mechanism in the coordinate representation, which is natural for many dynamical systems. The quantum amplitudes defining the mechanistic pathways are determined by a special procedure called coding Hamiltonians to access mechanistic pathways (CHAMP). Numerical procedures are presented for performing CHAMP calculations in a stable fashion. Mechanism determination is illustrated for excitation of a model linear triatomic molecule. Mechanisms are revealed for optimal control fields as well as for suboptimally driven systems including snapshots in time of the evolving mechanism and the effect of field noise on the mechanism. The high order multiphoton processes involved in the example produce many interfering pathways to create the final state, yet the dynamics and the mechanism are robust to reasonable levels of noise. PMID- 15332882 TI - Gaussian-type function set without prolapse 1H through 83Bi for the Dirac-Fock Roothaan equation. AB - We have developed prolapse free Gaussian basis sets which can be used for 1H to 83Bi, imposing the condition that the Dirac-Fock-Roothaan (DFR) total energy (TE) decreases monotonically toward the numerical DF (NDF) TE as the expansion term increases. An even-tempered basis set was assumed. The resulting sets gave |TE(DFR) - TE(NDF)| < or = 1 x 10(-6) hartree for any atoms less or equal to 83Bi; TE(NDF) = -21 565.638 345, and TE(DFR) = -21,565.638 345 +/- 0.000 001 hartree for Bi when the expansion terms are in the range (58, 58, 58, 36, 36, 36, and 36) and (72, 72, 72, 36, 36, 36, and 36) for (s+, p-, p+, d-, d+, f-, and f+) symmetries, respectively. A practical set with 44, 44, 44, 36, 36, 32, and 32 for the respective symmetries is also proposed where |TE(DFR) - TE(NDF)| < or = 4 x 10(-5). PMID- 15332883 TI - Altered inertial response of generic degrees of freedom. AB - A method to alter the inertial response of generic degrees of freedom is presented. A Hamiltonian acts on the inertial response of a set of independent configurational observables by the introduction of a kinetic bias. The method finds its natural application in reducing the spreading of arbitrary modes in a complex system. The resulting compressed spectrum allows a more efficient sampling of configurational space. PMID- 15332884 TI - Selective hyperfine excitation of N2H+ by He: potential energy surface, cross sections, and propensity rules. AB - We present potential energy surfaces for the He-N2H+ system adiabatically corrected for the zero-point motion along the intermolecular stretching vibrations v1 = 0 and v1 = 1. The potentials are extended to shorter He-N2H+ separations which makes them useful for scattering calculations. Close coupling calculations of the spinless S matrices for the rotational excitation of N2H+ by He are presented, and recoupling techniques to obtain collisional excitation cross sections between the N2H+ hyperfine levels are used. The propensity rules between hyperfine levels are investigated for the case where two nuclear spins are involved. It is found that the only well defined propensity rule is DeltaF = DeltaF1 = Deltaj and that calculations are required in order to obtain the relative intensities of the two-spin hyperfine cross sections. PMID- 15332885 TI - Femtosecond pump-probe spectroscopy of I2 in a dense rare gas environment: a mixed quantum/classical study of vibrational decoherence. AB - The process of decoherence of vibrational states of I2 in a dense helium environment is studied theoretically using the mixed quantum/classical method based on the Bohmian formulation of quantum mechanics [E. Gindensperger, C. Meier, and J. A. Beswick, J. Chem. Phys. 113, 9369 (2000)]. Specifically, the revival of vibrational wave packets is a quantum phenomena which depends sensitively on the coherence between the vibrational states excited by an ultrafast laser pulse. Its detection by a pump-probe setup as a function of rare gas pressure forms a very accurate way of detecting vibrational dephasing. Vibrational revivals of I2 in high pressure rare gas environments have been observed experimentally, and the very good agreement with the simulated spectra confirms that the method can accurately describe decoherence processes of quantum systems in interaction with an environment. PMID- 15332886 TI - Electron scattering from tetrafluoroethylene. AB - We report experimental results for electron scattering from tetrafluoroethylene, C2F4, obtained from measurements in two laboratories. An extensive set of differential, integral, and momentum transfer cross sections is provided for elastic scattering for incident electron energies from 1 to 100 eV and inelastic (vibrational excitation) scattering for incident electron energies at 3, 6, 7.5, 8, and 15 eV, and for scattering angles ranging from 10 degrees to 130 degrees. To highlight the role of intermediate negative ions (resonances) in the scattering process we have also measured excitation functions for elastic scattering and vibrational excitation of the ground electronic state of C2F4 for incident energies between 1.5 and 20 eV. Our results are compared with recent theoretical calculations and a limited number of other experimental results. PMID- 15332887 TI - Analysis of two intramolecular proton transfer processes in terms of the reaction force. AB - The negative derivative of the potential energy along an intrinsic reaction coordinate defines a force that has qualitatively a universal form for any process having an energy barrier: it passes through a negative minimum before the transition state, at which it is zero, followed by a positive maximum. We have analyzed two intramolecular proton transfer reactions in terms of several computed properties: internal charge separation, the electrostatic potentials of the atoms involved, their Fukui functions, and the local ionization energies. The variation of each of these properties along the intrinsic reaction coordinate shows a marked correlation with the characteristic features of the reaction force. We present a description of the proton transfer processes in terms of this force. PMID- 15332888 TI - Polarization spectroscopy of gaseous tropolone in a strong electric field. AB - We report studies of polarization spectroscopy of gaseous tropolone in a strong electric field using resonantly enhanced multiphoton ionization. The electric field induces localization of the tunneling proton between the two equivalent oxygen atoms. As a result, the C2v symmetry of the molecular frame is broken, and the parity selection rule is violated. The field induced transitions are type A with transition dipoles perpendicular to those under field free conditions. The polarization ratios, i.e., the ratios of the overall excitation yield under different polarizations of the resonant laser, thus deviate from those of a pure type B transition. In a field of 60 kV/cm, the experimental polarization ratio implies an essentially equal mixture of type B and type A transitions. Moreover, the induced transitions overlap with the two field-free subbands, and the resulting intensity ratios between the two subbands demonstrate dependence on the applied electric field. These observations can be qualitatively modeled using a quantum mechanical approach by assuming a two level system. A puzzling result is the magnitude of the transition dipole of the induced transition, which is proven to be essentially linearly dependent on the applied electric field. PMID- 15332889 TI - Theoretical study of excitations in furan: spectra and molecular dynamics. AB - The excitation spectra and molecular dynamics of furan associated with its low lying excited singlet states 1A2(3s), 1B2(V), 1A1(V'), and 1B1(3p) are investigated using an ab initio quantum-dynamical approach. The ab initio results of our previous work [J. Chem. Phys. 119, 737 (2003)] on the potential energy surfaces (PES) of these states indicate that they are vibronically coupled with each other and subject to conical intersections. This should give rise to complex nonadiabatic nuclear dynamics. In the present work the dynamical problem is treated using adequate vibronic coupling models accounting for up to four coupled PES and thirteen vibrational degrees of freedom. The calculations were performed using the multiconfiguration time-dependent Hartree method for wave-packet propagation. It is found that in the low-energy region the nuclear dynamics of furan is governed mainly by vibronic coupling of the 1A2(3s) and 1B2(V) states, involving also the 1A1(V') state. These interactions are responsible for the ultrafast internal conversion from the 1B2(V) state, characterized by a transfer of the electronic population to the 1A2(3s) state on a time scale of approximately 25 fs. The calculated photoabsorption spectrum of furan is in good qualitative agreement with experimental data. Some assignments of the measured spectrum are proposed. PMID- 15332890 TI - Accurate intermolecular ground state potential of the Ne-HCl van der Waals complex. AB - From an accurate ground state intermolecular potential energy surface we evaluate the rovibrational spectrum of the Ne-HCl van der Waals complex. The intermolecular potential is obtained by fitting a considerable number of interaction energies obtained at the coupled cluster singles and doubles including connected triple excitations level and with the augmented correlation consistent polarized valence quintuple zeta basis set extended with a set of 3s3p2d1f1g midbond functions. This basis set is selected after a systematic basis set study carried out at geometries close to those of the three main surface stationary points. The surface is characterized by two linear minima, i.e. Ne-ClH and Ne-HCl, with distances from the Ne atom to the HCl center of mass of 3.398 and 3.833 angstroms, respectively; and binding energies of -65.10 and -66.85 cm( 1), respectively. These results agree well with the experimental data available in contrast to previous theoretical results. The rovibrational spectra calculated for the different isotopic species are also compared to the experiments. PMID- 15332891 TI - Direct versus resonances mediated F+OH collisions on a new 3A" potential energy surface. AB - A theoretical study of the F(2P) + OH(2Pi) --> HF(1Sigma+) + O(3P) reactive collisions is carried out on a new global potential energy surface (PES) of the ground 3A" adiabatic electronic state. The ab initio calculations are based on multireference configuration interaction calculations, using the aug-cc-pVTZ extended basis sets of Dunning et al. A functional representation of the PES shows no nominal barrier to reaction, contrary to previous results by others. Wave packet and quasiclassical trajectory calculations have been performed for this PES to study the F + OH(v = 0,j) reactive collision. The comparison was performed at fixed and constant values of the total angular momentum from 0 to 110 and relative translational energy up to 0.8 eV. The reaction presents a dynamical barrier, essentially due to the zero-point energy for the bending vibration near the saddle point. This determines two different reaction mechanisms. At energies higher than approximately 0.125 eV the reaction is direct, while below that value it is indirect and mediated by heavy-light-heavy resonances. Such resonances, also found in the simulations of the photodetachment spectrum of the triatomic anion, manifest themselves in the quasiclassical simulations, too, where they are associated to periodic orbits. PMID- 15332892 TI - Electronic photodissociation spectroscopy of Aun- x Xe (n = 7-11) versus time dependent density functional theory prediction. AB - Electronic (one-photon) photodepletion spectra were recorded for gold cluster anions complexed with one xenon atom over the photon energy range 2.1-3.4 eV. Clusters were generated by pulsed laser vaporization and probed under collisionless molecular beam conditions. The spectra obtained are highly structured with the narrowest features--assigned to individual electronic transitions--having bandwidths of less than 40 meV. Time-dependent density functional theory predictions of optically allowed transitions for the most stable--planar--isomers of the corresponding bare metal cluster anions are generally consistent with the experimental observation. PMID- 15332893 TI - Between geometry, stability, and polarizability: density functional theory studies of silicon clusters Sin (n = 3-10). AB - The relationship between the polarizability, stability, and the geometry of small size silicon clusters has been investigated by the density functional theory methods. Results obtained at local density approximation/Vosko-Wilk-Nusair and general gradient approximation/BLYP levels with polarized even-tempered basis set of quadruple zeta quality are presented and compared with those obtained by the B3LYP method, as well as with the ab initio results in recent literature. We have found that the polarizability is directly related to the size of the energy gap between symmetry-compatible bonding and antibonding molecular orbitals, but not necessarily to the size of the HOMO-LUMO (highest occupied molecular orbital lowest unoccupied molecular orbital) gap. Furthermore, we have defined two structural parameters, namely, the averaged Si-Si distances and the standard deviation of the Si-Si distances, which were found to correlate remarkably well with the binding energy of the clusters and the HOMO-LUMO gap, respectively. These straightforward correlations would, therefore, provide a means to predict the physical properties, in particular, the polarizability and the stability, simply based on the structural information of the cluster. PMID- 15332894 TI - Ab initio calculations of stationary points on the benzene-Ar and p difluorobenzene-Ar potential energy surfaces: barriers to bound orbiting states. AB - The potential energy surfaces of the van der Waals complexes benzene-Ar and p difluorobenzene-Ar have been investigated at the second-order Moller-Plesset (MP2) level of theory with the aug-cc-pVDZ basis set. Calculations were performed with unconstrained geometry optimization for all stationary points. This study has been performed to elucidate the nature of a conflict between experimental results from dispersed fluorescence and velocity map imaging (VMI). The inconsistency is that spectra for levels of p-difluorobenzene-Ar and -Kr below the dissociation thresholds determined by VMI show bands where free p difluorobenzene emits, suggesting that dissociation is occurring. We proposed that the bands observed in the dispersed fluorescence spectra are due to emission from states in which the rare gas atom orbits the aromatic chromophore; these states are populated by intramolecular vibrational redistribution from the initially excited level [S. M. Bellm, R. J. Moulds, and W. D. Lawrance, J. Chem. Phys. 115, 10709 (2001)]. To test this proposition, stationary points have been located on both the benzene-Ar and p-difluorobenzene-Ar potential energy surfaces (PESs) to determine the barriers to this orbiting motion. Comparison with previous single point CCSD(T) calculations of the benzene-Ar PES has been used to determine the amount by which the barriers are overestimated at the MP2 level. As there is little difference in the comparable regions of the benzene-Ar and p difluorobenzene-Ar PESs, the overestimation is expected to be similar for p difluorobenzene-Ar. Allowing for this overestimation gives the barrier to movement of the Ar atom around the pDFB ring via the valley between the H atoms as < or = 204 cm(-1) in S0 (including zero point energy). From the estimated change upon electronic excitation, the corresponding barrier in S1 is estimated to be < or = 225 cm(-1). This barrier is less than the 240 cm(-1) energy of 30(2), the vibrational level for which the anomalous "free p-difluorobenzene" bands were observed in dispersed fluorescence from p-difluorobenzene-Ar, supporting our hypothesis for the origin of these bands. PMID- 15332895 TI - Electron scattering on OH-(H2O)n clusters (n = 0-4). AB - The cross sections for electron scattering on OH-(H2O)n for n = 0-4 were measured from threshold to approximately 50 eV. All detachment cross sections were found to follow the classical prediction given earlier [Phys. Rev. Lett. 74, 892 (1995)] with a threshold energy for electron-impact detachment that increased upon sequential hydration, yielding values in the range from 4.5 eV +/- 0.2 eV for OH- to 12.10 eV +/- 0.5 eV for OH-(H2O)4. For n > or = 1, we found that approximately 80% of the total reaction events lead to electron detachment plus total dissociation of the clusters into the constituent molecules of OH and H2O. Finally, we observed resonances in the cross sections for OH-(H2O)3 and for OH (H2O)4. The resonances were located at approximately 15 eV and were ascribed to the formation of dianions in excited states. PMID- 15332896 TI - Density functional theory study on the bridge structure in dimeric aluminum (III) water complexes. AB - Density-functional theory methods were used to investigate the structure of dimeric aluminum (III) water complexes as a function of bridging group. The possibilities of oxygen, water, and hydroxyl bridge ligands and a variety of structural arrangements, such as cis/trans, with respect to the relative position of hydroxyl ligands, were considered. Within the limit of our computational level, we found that electrostatic repulsion between hydroxyls is important in deciding the polyaluminum structure. Although the structures of aluminum-hexaaquo predominate, species with small number of charges or a large number of hydroxyl ligands have a tendency toward a five-coordinate trigonal bipyramidal configuration. Because water is electronically neutral, it cannot provide enough negative charges as a bridge ligand to stabilize two Al(III) molecules. The energy differences among many configurational isomers of hydroxyl Al are so small that they may coexist and convert into each other easily at room temperature. PMID- 15332897 TI - Structures, energetics, and spectra of hydrated hydroxide anion clusters. AB - The structures, energetics, electronic properties, and spectra of hydrated hydroxide anions are studied using density functional and high level ab initio calculations. The overall structures and binding energies are similar to the hydrated anion clusters, in particular, to the hydrated fluoride anion clusters except for the tetrahydrated clusters and hexahydrated clusters. In tetrahydrated system, tricoordinated structures and tetracoordinated structures are compatible, while in pentahydrated systems and hexahydrated systems, tetracoordinated structures are stable. The hexahydrated system is similar in structure to the hydrated chloride cluster. The thermodynamic quantities (enthalpies and free energies) of the clusters are in good agreement with the experimental values. The electronic properties induced by hydration are similar to hydrated chloride anions. The charge-transfer-to-solvent energies of these hydrated-hydroxide anions are discussed, and the predicted ir spectra are used to explain the experimental data in terms of the cluster structures. The low-energy barriers between the conformations along potential energy surfaces are reported. PMID- 15332898 TI - Dissolution of a base (RbOH) by water clusters. AB - Density functional and ab intio calculations are employed in order to understand the base dissociation of rubidium hydroxide by water molecules. The hydrated structures, stabilities, thermodynamic quantities, dissociation energies, infrared spectra, and electronic properties of RbOH(H2O)(n = 0-5) are investigated. With the successive addition of water molecules to RbOH, the Rb-OH bond lengthens significantly from 2.45 angstroms for n = 0 to 3.06 angstroms for n = 5. It is interesting to note that four water molecules are needed for the stable dissociation of RbOH (as an almost dissociate conformation) and five water molecules are needed for the complete dissociation without any Rb-OH stretch mode, in contrast to the same group base of CsOH which requires only three water molecules for an almost dissociate conformation and four water molecules for the complete dissociation. PMID- 15332899 TI - Rotational energy transfer in collisions between CO(X 1Sigma+, v= 2 , J = 0, 1, 4, and 6) and He at temperatures from 294 to 15 K. AB - Infrared-vacuum ultraviolet double resonance experiments have been implemented in the ultracold environment provided by a Cinetique de Reaction en Ecoulement Supersonique Uniforme apparatus. With this technique rate coefficients of two kinds have been measured for rotational energy transfer in collisions between CO and He: (a) those for total removal from the selected rotational states J = 0, 1, 4, and 6 in the vibronic state X 1Sigma+, v = 2, and (b) those for transfer between selected initial and specific final states. Using different Laval nozzles, results have been obtained at several different temperatures: 294, 149, 63, 27, and 15 K. The thermally averaged cross sections for total removal by collisions with He show only slight variations both with initial rotational state and with temperature. The variation of state-to-state rate coefficients with DeltaJ show several general features: (i) a decrease with increasing DeltaJ; (ii) a propensity to favor odd DeltaJ over even DeltaJ; and (iii) at lower temperatures, the distribution of rate coefficients against DeltaJ becomes narrower, and decreases in J are increasingly favored over increases in J, a preference which is most strongly seen for higher initial values of J. The results are shown to be in remarkably good agreement with those obtained in ab initio scattering calculations by Dalgarno and co-workers [Astrophys. J. 571, 1015 (2002)]. PMID- 15332900 TI - Photodissociation dynamics of 1,2-butadiene at 157 nm. AB - Photodissociation dynamics of 1,2-butadiene at 157 nm has been investigated using a molecular beam apparatus based on photoionization using vacuum ultraviolet synchrotron radiation. Six dissociation pathways have been observed. The observed channels are C4H5+H, C4H4+H2, C3H3+CH3, C2H3+C2H3, C2H4+C2H2, and C4H4+H+H. Among all the dissociation channels, the C3H3+CH3 channel is found to be the dominant process. The product kinetic energy distributions of all dissociation channels have been determined from simulating the experimental time-of-flight spectra. Relative branching ratios for all observed dissociation channels were also estimated based on all detected products. PMID- 15332901 TI - The potential energy surface of the Ar-CO complex obtained using high-resolution data. AB - A potential energy surface is retrieved for the Ar-CO complex by carrying out a global analysis of its high-resolution spectroscopic data. The data set consists of already published microwave and infrared data and of new microwave transitions which are presented in the paper. The theoretical approach used to reproduce the spectrum is based on a model Hamiltonian which accounts simultaneously for the two large amplitude van der Waals modes and for the overall rotation of the complex. Only the vCO = 0 state is considered. The root-mean-square deviation of the analysis is 18 MHz for the microwave data and 1.4 x 10(-3) cm(-1) for the infrared energy difference data. Fifteen parameters corresponding to the potential energy function are determined in addition to two kinetic energy parameters and two distortion-type parameters. The potential energy surface derived is in good agreement with the one obtained by Shin, Shin, and Tao [J. Chem. Phys. 104, 183 (1996)]. PMID- 15332902 TI - Structural stability and electronic state of transition metal trimers. AB - Ground state geometries were searched for transition metal trimers Sc3, Y3, La3, Lu3, Ti3, Zr3, and Hf3 by density functional methods. For all the studied trimers, our calculation indicates that the ground state geometries are either equilateral triangle (Zr3 and Hf3) or near equilateral triangle (Ti3, Sc3, Y3, La3, and Lu3). For rare earth trimers Sc3, Y3, La3, and Lu3, isosceles triangle (near equilateral triangle) at quartet state is the ground state. Isosceles triangle at doublet state is the competitive candidate for the ground state. For Zr3 and Hf3, equilateral triangle at singlet state is the most stable. For Ti3, isosceles triangle (near equilateral triangle) at quintet state gives the ground state. For Sc3, Zr3, and Hf3, where experimental results are available, the predicted geometries are in agreement with experiment in which the ground state is equilateral triangle (Zr3) or fluxional (Sc3 and Hf3). For Y3, the calculated geometry is in agreement with experimental observation and previous theoretical study that Y3 is a bent molecule for the ground state. For La3, our calculation is in excellent agreement with previous theoretical study based on density functional methods. PMID- 15332903 TI - Polarization effects on the hyper-Raman spectra of carbon tetrachloride: a joint experimental-theoretical study. AB - Hyper-Raman spectra of pure carbon tetrachloride in the liquid phase are recorded for different combinations of the polarizations of the incident and scattered lights and are compared to ab initio time-dependent Hartree-Fock simulations. Both the calculated intensities of the Raman and hyper-Raman spectra give indeed a quite satisfactory agreement with polarized experimental spectra. PMID- 15332904 TI - Structure changes in glassforming liquids upon cooling and compression. AB - Upon cooling and compression, both the thermodynamic and the kinetic properties of glassformers change. In fragile glassformers, these changes suggest changes in the local structure of the liquid. Thermodynamic data may then provide a measure of structure changes. Special criteria for structure changes in terms of energy and pressure fluctuations in the isochoric system are proposed; recent theoretical results allow one to rewrite the criteria in terms of long- and short time thermodynamic characteristics of the liquid. Based on the proposed criteria, we discuss the changes in the local structure due to temperature and pressure variations. PMID- 15332905 TI - Mass-fractal clustering and power-law decay of cluster size in 1-propanol aqueous solution. AB - Mesoscale structure of 1-propanol aqueous solutions with propanol mole fraction xp ranging from 0.1 to 0.33 has been studied by means of small angle neutron scattering (SANS) and large-scale reverse Monte Carlo (RMC) technique. Analysis of the SANS intensities in terms of a fractal model shows that the fractal dimension df of mesoscale structure of the solution is about 1.8-1.9 for water rich solution and about 1.5 for propanol-rich solution. Percolation analysis on the RMC results reveals that the water molecules and the propanol molecules cluster, respectively, as a mass fractal, the dimension dM of which is about 2.3 2.5 for both clusters for water-rich solution. Furthermore, the distribution of the cluster size is expressed by a simple power law with an exponent tau of about 1.35-1.5 for the propanol clusters and 1.05-1.2 for the water clusters. These results imply that the current solution is characterized by polydisperse mass fractals. In fact, a theoretical relation for polydisperse system of mass fractals, df = dM(2-tau), holds well in the current solution. The characteristic change in df from 1.8-1.9 to 1.5 described above is attributed to the crossover between the water-rich regime and the propanol-rich regime. Most of the water molecules and the propanol molecules are located on the interface between clusters, and the water molecules form thin layers of about 10 A thick irrespective of 1-propanol content studied. PMID- 15332906 TI - A discharge investigation of hydrogen and deuterium atom formation, and parahydrogen and orthodeuterium reconversion. AB - Hydrogen is flowed through a mild tesla-coil discharge and trapped at 3.8 K: New infrared absorptions of H2 are induced by interaction with trapped H atoms and H- anions. High purity parahydrogen and orthodeuterium samples are 1%-9% reconverted depending on the discharge pressure and recombination of atoms. Annealing the solid samples to 7 K reveals growth in p-H2 induced by o-H2, which shows that H atom recombination produces thermal nuclear spin populations. Similar results are found in discharge experiments with HD and on annealing solid HD. The observed increase in induced HD absorption by J = 1, H2 and D2 molecules formed on recombination gives approximately 1% for the H[D] atom concentration in our solid HD samples. PMID- 15332907 TI - Stark absorption spectroscopy of indole and 3-methylindole. AB - Indole and 3-methylindole (3-MI) doped into a polymethylmethacrylate (PMMA) film are studied by the Stark absorption (electroabsorption) spectroscopy. The 1La and 1Lb absorption bands are distinguished and the change in permanent dipole moment on 1La excitation is determined by a model fit to the measured absorption and electroabsorption spectra. Analysis of the spectra, measured at normal incidence and magic angle conditions, proved the essential role of the electric-field induced orientation/alignment effects for polar indole and 3-MI molecules in the PMMA environment at room temperature. PMID- 15332908 TI - Ab initio molecular-dynamics study of liquid formamide. AB - Properties of neat liquid formamide (HCONH2) have been studied by the combination of gradient-corrected density-functional theory, norm-conserving pseudopotentials, and the adaptive finite-element method. The structural and dynamical quantities have been calculated through molecular dynamics simulations under the Born-Oppenheimer approximation. Satisfactory agreement with experimental data was obtained for both intramolecular and intermolecular properties. Our results are also compared with those of the empirical potential functions to clarify their accuracies. PMID- 15332909 TI - Raman scattering study on structural and dynamical features of noncrystalline selenium. AB - We report on a detailed, temperature-dependent, off-resonant Raman scattering study of glassy and supercooled selenium. Raman spectra in the frequency regime of the first-order scattering (5-450 cm(-1)) have been recorded over a wide temperature range, i.e., 143-353 K. To facilitate the analysis, the spectra have intuitively been divided in three spectral regions. The analysis of the high frequency region (bond-stretching vibrational modes) yielded information on the rings-chains equilibrium. In particular, the polymer content was found to amount to more than 85% around the glass transition temperature, exhibiting a weak temperature dependence, which extrapolates nicely to the high-temperature dissolution data. The intermediate frequency range (representative of the medium range structural order) was treated together with the low frequency regime (where low-energy excitations, i.e., the quasielastic line and the Boson peak are the dominant contributions) owing to their strong overlap. The study of the bond bending regime revealed information which made it possible to clarify the role of ringlike and chainlike fragments incorporated in polymeric molecules. The temperature evolution of the Boson peak and the frequency dependence of the Raman coupling coefficient Comega were also determined. An attempt to decompose the partial contribution of the pure Boson peak to Comega revealed valuable information concerning the limiting (omega-->0) behavior of the coupling coefficient. PMID- 15332910 TI - Application of time series analysis on molecular dynamics simulations of proteins: a study of different conformational spaces by principal component analysis. AB - Time series analysis is applied on the collective coordinates obtained from principal component analysis of independent molecular dynamics simulations of alpha-amylase inhibitor tendamistat and immunity protein of colicin E7 based on the Calpha coordinates history. Even though the principal component directions obtained for each run are considerably different, the dynamics information obtained from these runs are surprisingly similar in terms of time series models and parameters. There are two main differences in the dynamics of the two proteins: the higher density of low frequencies and the larger step sizes for the interminima motions of colicin E7 than those of alpha-amylase inhibitor, which may be attributed to the higher number of residues of colicin E7 and/or the structural differences of the two proteins. The cumulative density function of the low frequencies in each run conforms to the expectations from the normal mode analysis. When different runs of alpha-amylase inhibitor are projected on the same set of eigenvectors, it is found that principal components obtained from a certain conformational region of a protein has a moderate explanation power in other conformational regions and the local minima are similar to a certain extent, while the height of the energy barriers in between the minima significantly change. As a final remark, time series analysis tools are further exploited in this study with the motive of explaining the equilibrium fluctuations of proteins. PMID- 15332911 TI - 2H nuclear magnetic resonance study of the molecular motion in cyanoadamantane. I. Supercooled plastically crystalline phase. AB - The supercooled plastically crystalline phase (glassy crystal) of cyanoadamantane was investigated by multidimensional 2H NMR (T>Tg). Although the orientationally disordered crystalline phase always coexisted with the orientationally ordered crystalline phase, we were able to single out the signal from the glassy crystal by selective excitation and it was possible to carry out line shape measurements and two-dimensional (2D) experiments (in frequency and time domain). The latter directly reveal sixfold jumps with an reorientation of the molecular C3 axis via 90 degrees angles, thus reflecting the symmetry of the lattice. The motion around the C3 axis is found to be always fast. We can reproduce the line shape by random walk simulations properly taking into account these molecular motions. Both methods (line shape and 2D experiments) yield time constants which agree with those reported by other techniques. Refining the analysis a narrow distribution of correlation times is introduced to account for a weak stretching of the correlation function. We did not find any indication of a small angle process usually found in structural glasses. Thus, the motional process in the glassy crystal appears to be simple and quite different from that in structural glasses. PMID- 15332912 TI - Single particle jumps in a binary Lennard-Jones system below the glass transition. AB - We study a binary Lennard-Jones system below the glass transition with molecular dynamics simulations. To investigate the dynamics we focus on events (jumps) where a particle escapes the cage formed by its neighbors. Using single particle trajectories we define a jump by comparing for each particle its fluctuations with its changes in average position. We find two kinds of jumps: "reversible jumps," where a particle jumps back and forth between two or more average positions, and "irreversible jumps," where a particle does not return to any of its former average positions, i.e., successfully escapes its cage. For all investigated temperatures both kinds of particles jump and both irreversible and reversible jumps occur. With increasing temperature, relaxation is enhanced by an increasing number of jumps and growing jump lengths in position and potential energy. However, the waiting time between two successive jumps is independent of temperature. This temperature independence might be due to aging, which is present in our system. We therefore also present a comparison of simulation data with three different histories. The ratio of irreversible to reversible jumps is also increasing with increasing temperature, which we interpret as a consequence of the increased likelihood of changes in the cages, i.e., a blocking of the "entrance" back into the previous cage. In accordance with this interpretation, the fluctuations both in position and energy are increasing with increasing temperature. A comparison of the fluctuations of jumping particles and nonjumping particles indicates that jumping particles are more mobile even when not jumping. The jumps in energy normalized by their fluctuations are decreasing with increasing temperature, which is consistent with relaxation being increasingly driven by thermal fluctuations. In accordance with subdiffusive behavior are the distributions of waiting times and jump lengths in position. PMID- 15332913 TI - Time resolved infrared absorption studies of geminate recombination and vibrational relaxation in OClO photochemistry. AB - Ultrafast time-resolved infrared absorption studies of aqueous chlorine dioxide (OClO) photochemistry are reported. Following photoexcitation at 401 nm, the evolution in optical density at frequencies between 1000 to 1100 cm(-1) is monitored to investigate vibrational energy deposition and relaxation along the asymmetric-stretch coordinate following the reformation of ground-state OClO via geminate recombination of the primary photofragments. The measured kinetics are compared to two proposed models for the vibrational-relaxation dynamics along the asymmetric-stretch coordinate. This comparison demonstrates that the perturbation model derived from molecular dynamics studies is capable of qualitatively reproducing the observed kinetics, where the collisional model employed in previous UV-pump, visible probe experiments demonstrates poor agreement with experiment. The ability of the perturbation model to reproduce the optical density evolution observed in these studies demonstrates that for aqueous OClO, frequency dependence of the solvent-solute coupling is important in defining the level-dependent vibrational relaxation rates along the asymmetric-stretch coordinate. The absence of optical-density evolution corresponding to the population of higher vibrational levels (n>8) along the asymmetric-stretch coordinate suggests that following geminate recombination, energy is initially deposited into a local Cl-O stretch, with the relaxation of vibrational energy from this coordinate providing for delayed vibrational excitation of the asymmetric- and symmetric-stretch coordinates relative to geminate recombination, as previously observed. PMID- 15332914 TI - Reconstruction of the free energy in the metastable region using the path ensemble. AB - By quenching into the metastable region of the three-dimensional Ising model, we investigate the paths that the magnetization (energy) takes as a function of time. We accumulate the magnetization (energy) paths into time-dependent distributions from which we reconstruct the free energy as a function of the magnetic field, temperature, and system size. From the reconstructed free energy, we obtain the free-energy barrier that is associated with the transition from a metastable state to the stable equilibrium state. Although mean-field theory predicts a sharp transition between the metastable and the unstable region where the free-energy barrier is zero, the results for the nearest-neighbor Ising model show that the free-energy barrier does not go to zero. PMID- 15332915 TI - Methyl rotational tunneling dynamics of p-xylene confined in a crystalline zeolite host. AB - The methyl rotational tunneling spectrum of p-xylene confined in nanoporous zeolite crystals has been measured by inelastic neutron scattering (INS) and proton nuclear magnetic resonance (NMR), and analyzed to extract the rotational potential energy surfaces characteristic of the methyl groups in the host-guest complex. The number and relative intensities of the tunneling peaks observed by INS indicate the presence of methyl-methyl coupling interactions in addition to the methyl-zeolite interactions. The INS tunneling spectra from the crystals (space group P2(1)2(1)2(1) with four crystallographically inequivalent methyl rotors) are quantitatively interpreted as a combination of transitions involving two coupled methyl rotors as well as a transition involving single-particle tunneling of a third inequivalent rotor, in a manner consistent with the observed tunneling energies and relative intensities. Together, the crystal structure and the absence of additional peaks in the INS spectra suggest that the tunneling of the fourth inequivalent rotor is strongly hindered and inaccessible to INS measurements. This is verified by proton NMR measurements of the spin-lattice relaxation time which reveal the tunneling characteristics of the fourth inequivalent rotor. PMID- 15332916 TI - Selective bonding of pyrazine to silicon(100)-2x1 surfaces: the role of nitrogen atoms. AB - The covalent binding of pyrazine on Si(100) have been investigated using high resolution electron energy loss spectroscopy (HREELS) and x-ray photoelectron spectroscopy. Experimental results clearly suggest that the attachment occurs exclusively through the bonding of the two para-nitrogen atoms with the surface without the involvement of the carbon atoms, as evidenced from the retention of the (sp2) C-H stretching mode in HREELS and a significant down shift of 1.6 eV in the binding energy of N 1s. The binding mechanism for pyrazine on Si(100) demonstrates that reaction channels for heteroatomic aromatic molecules are strongly dependent on the electronic properties of the constituent atoms. PMID- 15332917 TI - Simulation of the mechanical unfolding of ubiquitin: probing different unfolding reaction coordinates by changing the pulling geometry. AB - Motivated by the recent experimental atomic force microscopy (AFM) measurements of the mechanical unfolding of proteins pulled in different directions [D. J. Brockwell et al., Nat. Struct. Biol. 10, 731 (2003); M. Carrion-Vazquez et al., ibid 10, 738 (2003)] we have computed the unfolding free energy profiles for the ubiquitin domain when it is stretched between its (A) N and C termini, (B) Lys48 and C terminus, (C) Lys11 and C terminus, and (D) N terminus and Lys63. Our results for cases (A) and (B) are in good agreement with the experimental unfolding forces measured for the N-C and Lys48-C linked polyubiquitin, in particular, indicating a considerably lower unfolding force in the latter case. Mechanical unfolding in case (A) involves longitudinal shearing of the terminal parallel strands while in case (C) the same strands are "unzipped" by the force. The computed unfolding forces in case (C) are found to be very low, less than 50 pN for pulling rates typical of AFM experiments. The unfolding free energy barrier found in case (C) is approximately 13 kcal/mol, which corresponds to a zero-force unfolding rate constant that is comparable to the rate of chemical unfolding extrapolated to zero denaturant concentration. The unfolding barrier calculated in case (A) in the limit of zero force is much higher, suggesting that mechanical unfolding in this case follows a pathway that is different from that of thermal/chemical denaturation. PMID- 15332918 TI - Low-energy impact of X-(H2O)n (X = Cl,I) onto solid surface. AB - We investigated dissociation of X-(H2O)n (X = Cl, I, n = 13-31) by the impact onto a (La0.7Ce0.3)B6(100) surface at a collision energy Ecol of 1-5 eV per water molecule in a tandem time-of-flight mass spectrometer equipped with a translation energy analyzer. The mechanism of the dissociation was elucidated on the basis of the measurements of the mass spectrum and the translational energies of the product anions, X-(H2O)m (m = 0-4), scattered from the surface. It was concluded that (1) the parent cluster anion impacted on the surface undergoes dissociation on the surface under quasiequilibrium with its characteristic time varying with Ecol and n, and (2) the total collision energy introduced is partitioned preferentially to the translational motions of the products on the surface and to the rotational, the vibrational, and the lattice vibrational motions (surface) in this order. The quasiequilibrium model is applicable, even at the collision energy as low as 1 eV, because the translational modes are found to be statistically distributed while the other modes are not much populated by dynamical and energetics limitation. PMID- 15332919 TI - Vibrational dynamics of adsorbed molecules under conditions of photodesorption: pump-probe SFG spectra of CO/Pt(111). AB - Interaction of CO adsorbed on Pt(111) with electrons and phonons is studied experimentally by means of a pump-probe experiment where CO is probed by IR + visible sum frequency generation under a pump laser intensity that allows photodesorption. Vibrational spectra of CO internal stretch are obtained as a function of pump-probe delay. A two-temperature and anharmonic coupling model is used to extract from the spectra the real time variations of CO peak frequency and dephasing time. The main conclusions are the following: (i) The CO stretch is perturbed by two low-frequency modes, assigned to frustrated rotation and frustrated translation. (ii) The frustrated rotation is directly coupled to electrons photoexcited in Pt(111) by the pump laser. (iii) There is no evidence of Pt-CO stretch excitation in the spectra. The implications for the photodesorption dynamics are discussed. PMID- 15332921 TI - Polymer adsorption onto random planar surfaces: interplay of polymer and surface correlations. AB - We study the adsorption of homogeneous or heterogeneous polymers onto heterogeneous planar surfaces with exponentially decaying site-site correlations, using a variational reference system approach. As a main result, we derive simple equations for the adsorption-desorption transition line. We show that it is preferable to have a small amount of strongly adsorbing sites or monomers rather than a greater amount of weakly adsorbing ones. The results are discussed with respect to their implications for the physics of molecular recognition. PMID- 15332920 TI - X-ray emission spectroscopy of (2 square root of 3 x 2 square root of 3)R30 degrees CO/Ru(0001): comparison to c(2x2)CO/Ni(100) and c(2x2)CO/Cu(100). AB - The atom specific electronic structure of (2 square root of 3 x 2 square root of 3)R30 degrees CO on hcp Ru(0001) has been determined with resonantly excited x ray emission spectroscopy. We find that the general features of the local adsorbate electronic structure are similar to the situation of CO adsorbed on the fcc metals Ni(100) and Cu(100). The interpretation of the surface chemical bond of (2 square root of 3 x 2 square root of 3)R30 degrees CO/Ru(0001) based on the direct application of the local, allylic model from on-top adsorption on the fcc(100) surfaces Ni(100) and Cu(100) explains many aspects of the surface chemical bond. However, also nonlocal contributions like adsorbate-adsorbate interaction and the deviation from upright on-top adsorption on the Ru(0001) surface influence observables like the heat of adsorption and the Me-CO bond strength. PMID- 15332922 TI - Structure of short polymers at interfaces: a combined simulation and theoretical study. AB - The structure of polymers confined between surfaces is studied using computer simulation and a density functional approach. The simple model system considers the polymer molecule as a pearl necklace of freely jointed hard spheres, having attractions among the beads, confined between attractive surfaces. This approach uses the universality of the free-energy functional to obtain the self-consistent field required in the single chain simulation. The second-order direct correlation function for the uniform bulk fluid required as input has been calculated from the reference interaction site model integral equation theory using mean spherical approximation. The theoretical results are shown to compare well with the Monte Carlo simulation results for varying densities, chain lengths, and with different attractive interaction parameters. The simulation results on the conformational properties give important indications regarding the behavior of chains as they approach the surfaces. PMID- 15332923 TI - Surface ordering of diskotic liquid crystals. AB - We present Monte Carlo simulations of diskotic molecules using the Gay-Berne potential in a slab geometry. The disk-wall interaction is described by two different functions according to whether or not the equilibrium distance is dependent on the relative orientation of the disk to the wall. Furthermore, by changing the parameters of these potentials, we model either homeotropic (face on) or planar (edge-on) anchoring of the disks. We have found that the isotropic nematic transition does not change in comparison with the bulk situation. The temperature of the nematic-columnar transition, on the contrary, is found to increase for homeotropic anchoring, and decrease for planar anchoring, independently of the details of the potential. We explain the decrease of the transition temperature in the planar anchoring situation as the result of an induced frustration, due to the competition between the two orientations induced independently by the upper and lower walls. PMID- 15332924 TI - Trapping time statistics and efficiency of transport of optical excitations in dendrimers. AB - We theoretically study the trapping time distribution and the efficiency of the excitation energy transport in dendritic systems. Trapping of excitations, created at the periphery of the dendrimer, on a trap located at its core, is used as a probe of the efficiency of the energy transport across the dendrimer. The transport process is treated as incoherent hopping of excitations between nearest neighbor dendrimer units and is described using a rate equation. We account for radiative and nonradiative decay of the excitations while diffusing across the dendrimer. We derive exact expressions for the Laplace transform of the trapping time distribution and the efficiency of trapping, and analyze those for various realizations of the energy bias, number of dendrimer generations, and relative rates for decay and hopping. We show that the essential parameter that governs the trapping efficiency is the product of the on-site excitation decay rate and the trapping time (mean first passage time) in the absence of decay. PMID- 15332925 TI - Energy density analysis of cluster size dependence of surface-molecule interactions: H2, C2H2, C2H4, and CO adsorption onto Si(100)-(2x1) surface. AB - Adsorption of H2, C2H2, C2H4, and CO onto a Si(100)-(2x1) surface has been treated theoretically using Si(12n - 3)H(8n + 4) (n = 1-4) clusters. The energy density analysis (EDA) proposed by Nakai has been adopted to examine surface molecule interactions for different cluster sizes. EDA results for the largest model cluster Si45H36 have shown that the adsorption-induced energy density variation in Si atoms decays with distance from the adsorption site. Analysis of this decay, which can be carried out using the EDA technique, is important because it enables verification of the reliability of the model cluster used. In the cases of H2, C2H2, C2H4, and CO adsorption onto the Si(100)-(2x1) surface, it is found that at least a Si21H20 cluster is necessary to treat the surface molecule interaction with chemical accuracy. PMID- 15332926 TI - Dual control cell reaction ensemble molecular dynamics: a method for simulations of reactions and adsorption in porous materials. AB - We present a simulation tool to study fluid mixtures that are simultaneously chemically reacting and adsorbing in a porous material. The method is a combination of the reaction ensemble Monte Carlo method and the dual control volume grand canonical molecular dynamics technique. The method, termed the dual control cell reaction ensemble molecular dynamics method, allows for the calculation of both equilibrium and nonequilibrium transport properties in porous materials such as diffusion coefficients, permeability, and mass flux. Control cells, which are in direct physical contact with the porous solid, are used to maintain the desired reaction and flow conditions for the system. The simulation setup closely mimics an actual experimental system in which the thermodynamic and flow parameters are precisely controlled. We present an application of the method to the dry reforming of methane reaction within a nanoscale reactor model in the presence of a semipermeable membrane that was modeled as a porous material similar to silicalite. We studied the effects of the membrane structure and porosity on the reaction species permeability by considering three different membrane models. We also studied the effects of an imposed pressure gradient across the membrane on the mass flux of the reaction species. Conversion of syngas (H2/CO) increased significantly in all the nanoscale membrane reactor models considered. A brief discussion of further potential applications is also presented. PMID- 15332927 TI - Soft effective interactions between weakly charged polyelectrolyte chains. AB - We apply extensive molecular dynamics simulations and analytical considerations in order to study the conformations and the effective interactions between weakly charged, flexible polyelectrolyte chains in salt-free conditions. We focus on charging fractions lying below 20%, for which case there is no Manning condensation of counterions and the latter can be thus partitioned in two states: those that are trapped within the region of the flexible chain and the ones that are free in the solution. We examine the partition of counterions in these two states, the chain sizes and the monomer distributions for various chain lengths, finding that the monomer density follows a Gaussian shape. We calculate the effective interaction between the centers of mass of two interacting chains, under the assumption that the chains can be modeled as two overlapping Gaussian charge profiles. The analytical calculations are compared with measurements from molecular dynamics simulations. Good quantitative agreement is found for charging fractions below 10%, where the chains assume coil-like configurations, whereas deviations develop for charge fraction of 20%, in which case a conformational transition of the chain towards a rodlike configuration starts to take place. PMID- 15332928 TI - Water network dynamics at the critical moment of a peptide's beta-turn formation: a molecular dynamics study. AB - All-atom molecular dynamics simulations for a single molecule of Leu-Enkephalin in aqueous solution have been used to study the role of the water network during the formation of beta-turns. We give a detailed account of the intramolecular hydrogen bonding, the water-peptide hydrogen bonding, and the orientation and residence times of water molecules focusing on the short critical periods of transition to the stable beta-turns. These studies suggest that, when intramolecular hydrogen bonding between the first and fourth residue of the beta turn is not present, the disruption of the water network and the establishment of water bridges constitute decisive factors in the formation and stability of the beta-turn. Finally, we provide possible explanations and mechanisms for the formations of different kinds of beta-turns. PMID- 15332929 TI - On the phase diagram of reentrant condensation in polyelectrolyte-liposome complexation. AB - Complexation of polyions with oppositely charged spherical liposomes has been investigated by means of dynamic light scattering measurements and a well-defined reentrant condensation has been observed. The phase diagram of charge inversion, recently derived [T. T. Nguyen and B. I. Shklovskii, J. Chem. Phys. 115, 7298 (2001)] for the complexation of DNA with charged spherical macroions, has been employed in order to define the boundaries of the region where polyion-liposome complexes begin to condense, forming larger aggregates, and where aggregates dissolve again, towards isolated polyion-coated-liposome complexes. A reasonable good agreement is observed in the case of complexes formed by negatively charged polyacrylate sodium salt polyions and liposomes built up by cationic lipids (dioleoyltrimethylammoniumpropane), in an extended liposome concentration range. PMID- 15332930 TI - Simulation of the plastic behavior of amorphous glassy bis-phenol-A polycarbonate. AB - A protocol for studying the plastic deformation of amorphous glassy polymers is presented. The protocol is based on a viable computational procedure which combines constant-stress molecular dynamics simulations and fixed-cell energy minimizations, followed by kinetic, configurational, and energy analyses. It is shown that the computational results can be accounted for within a "potential energy landscape" theoretical framework, in which the plastic transition is interpreted as a crossing between and a collapse onto each other of "ideal (thermodynamic) structures." The procedure is applied to bis-phenol-A polycarbonate (BPA-PC), but is equally valid for a wide variety of polymeric species. Allowing for the limited size of the simulation cell, the high strain rate, and the fact that the simulation are conducted at low temperature, the values of the density, Young's modulus, yield strain, yield stress, activation energy, and activation volume are in fair agreement with the experimental data on BPA-PC. The analysis of the results shows that the plastic relaxation for this polymer has both a collective and cooperative character (as in classical percolation theories), involves a significant fraction of the simulation cell, and can be viewed as a "nanoscopic shear band." PMID- 15332932 TI - A molecular dynamics simulation study of the alpha-relaxation in a 1,4 polybutadiene melt as probed by the coherent dynamic structure factor. AB - The dynamic coherent structure factor Scoh(q,t) for a 1,4-polybutadiene (PBD) melt has been investigated using atomistic molecular dynamics simulations. The relaxation of Scoh(q,t) at q = 1.44 angstroms(-1) and q = 2.72 angstroms(-1), corresponding to the first and second peaks in the static structure factor for PBD, was studied in detail over a wide range of temperature. It was found that time-temperature superposition holds for the alpha-relaxation for both q values over a wide temperature range and that the alpha-relaxation can be well described by a stretched (Kohlrauch-William-Watts) exponential with temperature independent but q dependent amplitude and stretching exponent. The alpha-relaxation times for both q values were found to exhibit the same non-Arrhenius temperature dependence, indicating that the same physical processes are responsible for relaxation on both length scales. The alpha-relaxation time was found to depend strongly upon the dynamical range of data utilized in determining the relaxation time, accounting for qualitative discrepancies between alpha-relaxation times reported here and those extracted for PBD from experimentally measured Scoh(q,t). PMID- 15332931 TI - Modeling of intramolecular reactions of polymers: an efficient method based on Brownian dynamics simulations. AB - By the traditional approach to the Brownian dynamics simulations of intrachain reactions of polymers, the initial chain conformation is sampled from the equilibrium distribution. A dynamic trajectory is carried out until a "collision" of the reactive groups takes place, i.e., the distance between their centers becomes less that a certain reaction radius. The average length of the trajectory is equal to the mean time tauF of a diffusion-controlled reaction. In this work we propose another computational scheme. The trajectory begins at the instant of collision and is carried out until the chain is relaxed. The length of the trajectory has the order of the relaxation time taurel of the distance between the reactive groups. For polymer systems with taurel << tauF, this scheme allows the computation of tauF with considerable gain in computational time. Using the present approach, we calculated the mean time of DNA cyclization for the molecule length in the range from 50 to 500 nm. PMID- 15332933 TI - A theory of polymer solutions without the mean-field approximation in Flory Huggins theory. AB - The concept of the volume fraction at chain end is proposed, which is the conditional probability for a site having been occupied, knowing that an adjacent site is occupied by polymer end. The overall entropy of polymer/solvent system is separated into two fundamentally different parts, i.e., the translational entropy and the conformational entropy. Based on these a theory of polymer solutions is established. When a mean-field approximation is introduced, Flory-Huggins (FH) theory is recovered. The FH interaction parameters and spinodal curves of the polystyrene/cyclohexane system are calculated and compared with the experimental data. The good prediction of them two is achieved. PMID- 15332934 TI - Continuous polydispersity in a self-consistent field theory for diblock copolymers. AB - An efficient algorithm is presented for numerically evaluating a self-consistent field theoretic (SCFT) model of an AB diblock copolymer that incorporates continuous polydispersity in one of the blocks. An interesting segregation effect is found in which chains of intermediate molecular weight are concentrated at domain interfaces. This model of continuous polydispersity is also implemented in the random phase approximation (RPA) to study the order-disorder transition and predicts that the stability of the disordered, homogeneous phase decreases as the polydispersity in one of the blocks increases. The RPA predictions are confirmed by SCFT calculations. Our approach and results are particularly relevant to block copolymers prepared by quasiliving synthesis techniques, where the polymerization of one block is much more controlled than the other block. PMID- 15332935 TI - Interactions between colloidal particles in polymer solutions: A density functional theory study. AB - We present a density functional theory study of colloidal interactions in a concentrated polymer solution. The colloids are modeled as hard spheres and polymers are modeled as freely jointed tangent hard sphere chains. Our theoretical results for the polymer-mediated mean force between two dilute colloids are compared with recent simulation data for this model. Theory is shown to be in good agreement with simulation. We compute the colloid-colloid potential of mean force and the second virial coefficient, and analyze the behavior of these quantities as a function of the polymer solution density, the polymer chain length, and the colloid/polymer bead size ratio. PMID- 15332936 TI - Micellization behavior of star-block copolymers in a selective solvent: A Brownian dynamics simulation approach. AB - Micellization behavior of (AB)n type star-block copolymer in a selective solvent for its outer block is investigated by using a Brownian dynamics simulation. Micellar properties are compared in terms of the arm number (n) of star-block copolymer. It is observed that the critical micelle concentration (cmc) shows a minimum when the cmc is plotted against the arm number. The star-block copolymer with longer soluble block shows the cmc minimum at smaller arm number than that with shorter soluble block. Although the star-block copolymer with multiple arms forms more compact core as compared to diblock copolymer, the average aggregation number is inversely proportional to the arm number (approximately 1/n), which implies that the micelle size is invariant with the arm number. Theoretical predictions based on a simple mean field theory agree qualitatively well with the simulation results. PMID- 15332937 TI - Classical density functional study of mixed amphiphile mesostructures. AB - This paper presents a density functional study of mixed amphiphile solutions. The amphiphiles are modeled using four to seven fused hard spheres where the connectivity is such that the amphiphiles have a proper head group. We present calculations of lamellar and vesicle density distributions, paying particular attention to the morphology of the bilayer structures. PMID- 15332938 TI - The orientational order parameters of a dendritic liquid crystal organo-siloxane tetrapode oligomer, determined using polarized infrared spectroscopy. AB - The observed macroscopic anisotropic properties such as the components of infrared (IR) absorbances of liquid crystals are expressed in terms of the order parameters of the long molecular axis, molecular, and phase biaxiality. The order parameters of the organo-siloxane tetrapode liquid crystal of zero dendritic order (G0) in its nematic and smectic phases have been determined using results of the polarized IR spectroscopic measurements on a planar homogenously and hometropic aligned cells. The spatial components of the absorbances for the vibrational bands (in the mesogenic unit, terminal chains, and spacer) have been measured and analyzed. For the laboratory reference system, the apparent orientational order parameter S of the mesogen unit shows a significant drop in the transition from the nematic to the smectic phase while the phase biaxiality order parameter P increases to almost 0.4 in the smectic phase. This result shows that the director is tilted out of the sample plane in the smectic phase. The molecular biaxiality parameter D is found to be positive both for the nematic and smectic phases. This suggests that the carbonyl dipoles are oriented close to the tilt plane. For the vibrational bands in the chains, low values of S and D indicative of their low orientational order are obtained. As a result of the interaction among the molecules in the tilted smectic phases, the transition dipoles show positive correlations for the transversal and negative for the longitudinal dipoles. PMID- 15332940 TI - Quantum amplifier: measurement with entangled spins. AB - It is experimentally demonstrated that entangled quantum states can be used to amplify perturbations and to increase changes in observable values. The physical system is seven nuclear spins of single-labeled (13)C-benzene in a liquid crystalline matrix. An entangled state of six proton spins was used to monitor interaction with the (13)C spin. PMID- 15332941 TI - Calculation of current densities using gauge-including atomic orbitals. AB - A method for calculating the various components of the magnetically induced current-density tensor using gauge-including atomic orbitals is described. The method is formulated in the framework of analytical derivative theory, thus enabling implementation at the Hartree-Fock self-consistent-field (HF-SCF) as well as at electron-correlated levels. First-order induced current densities have been computed up to the coupled-cluster singles and doubles level (CCSD) augmented by a perturbative treatment of triple excitations [CCSD(T)] for carbon dioxide and benzene and up to the full coupled-cluster singles, doubles, and triples (CCSDT) level in the case of ozone. The applicability of the gauge including magnetically induced current method to larger molecules is demonstrated by computing first-order current densities for porphin and hexabenzocoronene at the HF-SCF and density-functional theory level. Furthermore, a scheme for obtaining quantitative values for the induced currents in a molecule via numerical integration over the current flow is presented. For benzene, a perpendicular magnetic field induces a (field dependent) ring current of 12.8 nA T(-1) at the HF-SCF level using a triple-zeta basis set augmented with polarization functions (TZP). At the CCSD(T)/TZP level the induced current was found to be 11.4 nA T(-1). Gauge invariance and its relation to charge-current conservation is discussed. PMID- 15332942 TI - Simple minimum principle to derive a quantum-mechanical/molecular-mechanical method. AB - We propose a minimum principle to derive a QM/MM (quantum-mechanical/molecular mechanical) method from the first principle. We approximate the Hamiltonian of a spectator substituent as the structure-dependent effective Hamiltonian in a least squares sense. This effective Hamiltonian is expanded with the orthogonal operator set called the normal-ordered product. We determine the structure dependent energy that corresponds to the classical MM energy and the extra one electron potential that takes account of the interface effects. This QM/MM method is free from the double-counting problem and the artificial truncation of the localized molecular orbitals. As a numerical example we determine the one electron effective Hamiltonian of the methyl group. This effective Hamiltonian is applied to the ethane and CH(3)CH(2)X molecules (X=CH(3), NH(2), OH, F, COOH, NH(3) (+), OH(2) (+), and COO(-)). It reproduced the relative energies, potential energy curves, and the Mulliken populations of the all-electron calculations fairly well. PMID- 15332943 TI - Quantum corrections to classical time-correlation functions: hydrogen bonding and anharmonic floppy modes. AB - Several simple quantum correction factors for classical line shapes, connecting dipole autocorrelation functions to infrared spectra, are compared to exact quantum data in both the frequency and time domain. In addition, the performance of the centroid molecular dynamics approach to line shapes and time-correlation functions is compared to that of these a posteriori correction schemes. The focus is on a tunable model that is able to describe typical hydrogen bonding scenarios covering continuously phenomena from tunneling via low-barrier hydrogen bonds to centered hydrogen bonds with an emphasis on floppy modes and anharmonicities. For these classes of problems, the so-called "harmonic approximation" is found to perform best in most cases, being, however, outperformed by explicit centroid molecular dynamics calculations. In addition, a theoretical analysis of quantum correction factors is carried out within the framework of the fluctuation dissipation theorem. It can be shown that the harmonic approximation not only restores the detailed balance condition like all other correction factors, but that it is the only one that also satisfies the fluctuation-dissipation theorem. Based on this analysis, it is proposed that quantum corrections of response functions in general should be based on the underlying Kubo-transformed correlation functions. PMID- 15332944 TI - Path integral approach to Brownian motion driven with an ac force. AB - Brownian motion in a periodic potential driven by an ac (oscillatory) force is investigated for the full range of damping constant from the overdamped limit to the underdamped limit. The path (functional) integral approach is advanced to produce formulas for the probability distribution function and for the current of the Brownian particle in response to an ac driving force. The negative friction Langevin dynamics technique is employed to evaluate the dc current for various parameters without invoking the overdamped or the underdamped approximation. The dc current is found to have nonlinear dependence upon the damping constant, the potential parameter, and the ac force magnitude and frequency. PMID- 15332945 TI - A quantum chemical approach to the free energy calculations in condensed systems: the QM/MM method combined with the theory of energy representation. AB - A methodology has been proposed to compute the solvation free energy of a molecule described quantum chemically by means of quantum mechanical/molecular mechanical method combined with the theory of energy representation (QM/MM-ER). The present approximate approach is quite simple to implement and requires much less computational cost as compared with the free energy perturbation or thermodynamic integration. Furthermore, the electron distribution can be treated faithfully as a quantum chemical object, and it is no longer needed to employ the artificial interaction site model, a reduced form of the realistic electron distribution, which is commonly used in the conventional solution theory. The point of the present approach is to employ the QM solute with electron density fixed at its average distribution in order to make the solute-solvent interaction pairwise. Then, the solvation free energy can be computed within the standard framework of the energy representation. The remaining minor contribution originating from the many-body effect inherent in the quantum mechanical description can be evaluated separately within a similar framework if necessary. As a test calculation, the method has been applied to a QM water solute solvated by MM water solvent in ambient and supercritical states. The results of the QM/MM ER simulations have been in excellent agreement with the experimental values. PMID- 15332946 TI - A field theoretical approach to calculate electronic Born-Oppenheimer coupling terms. AB - In this paper we suggest to consider the spatial distribution of the Born Oppenheimer nonadiabatic coupling terms as fields which are created by sources, located at degeneracy points, and which can be derived using the ordinary mathematical tools of field theory. It is shown that the curl-divergence equations as formed within a given Hilbert space [M. Baer, Chem. Phys. Lett. 35, 112 (1975)] can be converted into a set of inhomogeneous coupled Poisson equations which are solved for a given set of boundary conditions. The method is applied to the three-state Hilbert subspace of the H(3) system. The numerical results are compared with ab initio calculations for which a very encouraging fit is found. PMID- 15332947 TI - Theoretical study of the electronic states of Nb4, Nb5 clusters and their anions (Nb4-,Nb5-). AB - Geometries and energy separations of the various low-lying electronic states of Nb(n) and Nb(n) (-) (n=4,5) clusters with various structural arrangements have been investigated. The complete active space multiconfiguration self-consistent field method followed by multireference singles and doubles configuration interaction (MRSDCI) calculations that included up to 52x10(6) configuration spin functions have been used to compute several electronic states of these clusters. The ground states of both Nb(4) ((1)A('), pyramidal) and Nb(4) (-) ((2)B(3g), rhombus) are low-spin states at the MRSDCI level. The ground state of Nb(5) cluster is a doublet with a distorted trigonal bipyramid (DTB) structure. The anionic cluster of Nb(5) has two competitive ground states with singlet and triplet multiplicities (DTB). The low-lying electronic states of these clusters have been found to be distorted due to Jahn-Teller effect. On the basis of the energy separations of our computed electronic states of Nb(4) and Nb(5), we have assigned the observed photoelectron spectrum of Nb(n) (-) (n=4,5) clusters. We have also compared our MRSDCI results with density functional calculations. The electron affinity, ionization potential, dissociation and atomization energies of Nb(4) and Nb(5) have been calculated and the results have been found to be in excellent agreement with the experiment. PMID- 15332948 TI - Potential energy constrained molecular dynamics simulations. AB - A method for carrying out molecular dynamics simulations in which the potential energy U of the molecular system is constrained at its initial value is developed and thoroughly tested. The constraint is not introduced within the framework of the Lagrange multipliers technique, rather it is fulfilled in a natural way by carrying out the simulations in terms of suitable sets of delocalized coordinates. Such coordinates are defined by an appropriate tuning of the Baker, Kessi, and Delley internal delocalized nonredundant coordinates technique [J. Chem. Phys. 105, 192 (1996)]. The proposed method requires multiple evaluations of energy and gradients in each step of the molecular dynamics simulation, so that constant U simulations suffer some overhead compared to ordinary simulations. But the particular formulation of the delocalized coordinates and of the equations of motion greatly simplifies all the various steps required by the Baker's technique, thus allowing for the efficient implementation of the method itself. The technique is reliable and allows for very high accuracy in the potential energy conservation during the whole simulation. Moreover, it proved to be free of drift troubles which can occur when standard constraint methods are straightforwardly implemented without the application of appropriate correcting techniques. PMID- 15332949 TI - A quasidegenerate formulation of the second order n-electron valence state perturbation theory approach. AB - The n-electron valence state perturbation theory (NEVPT) is reformulated in a quasidegenerate (QD) approach. The new theory allows the treatment of cases where the proximity of the energies causes artifacts in the zero order description. Problems of quasidegeneration are relevant in the dynamics involving regions at avoided crossings (or conical intersections) and in spectroscopy where the energies and oscillator strengths can be strongly influenced by the mixing of states of different nature. Two test cases are analyzed concerning (a) the ionic neutral avoided crossing in LiF and (b) the valence/Rydberg mixing in the excited states of ethene. The QD-NEVPT2 is shown to be a useful tool for such systems. PMID- 15332950 TI - New recurrence relations for the rapid evaluation of electron repulsion integrals based on the accompanying coordinate expansion formula. AB - We present an algorithm for the rapid computation of electron repulsion integrals (ERIs) over Gaussian basis functions based on the accompanying coordinate expansion (ACE) formula. The present algorithm uses equations termed angular momentum reduced expressions and introduces two types of recurrence relations to ACE formulas. Numerical efficiencies are assessed for (p pmid R:p p) and (sp spmid R:sp sp) ERIs by using the floating-point operation count. The algorithm is suitable for calculating ERIs for the same exponents but different angular momentum functions, such as L shells and derivatives of ERIs. The present algorithm is also capable of calculating ERIs with highly contracted Gaussian basis functions. PMID- 15332951 TI - Efficient formulation of the stochastic simulation algorithm for chemically reacting systems. AB - In this paper we examine the different formulations of Gillespie's stochastic simulation algorithm (SSA) [D. Gillespie, J. Phys. Chem. 81, 2340 (1977)] with respect to computational efficiency, and propose an optimization to improve the efficiency of the direct method. Based on careful timing studies and an analysis of the time-consuming operations, we conclude that for most practical problems the optimized direct method is the most efficient formulation of SSA. This is in contrast to the widely held belief that Gibson and Bruck's next reaction method [M. Gibson and J. Bruck, J. Phys. Chem. A 104, 1876 (2000)] is the best way to implement the SSA for large systems. Our analysis explains the source of the discrepancy. PMID- 15332952 TI - The extended Perdew-Burke-Ernzerhof functional with improved accuracy for thermodynamic and electronic properties of molecular systems. AB - Density functional theory (DFT) has become the method of choice for many applications of quantum mechanics to the study of the electronic properties of molecules and solids. Despite the enormous progress in improving the functionals, the current generation is inadequate for many important applications. As part of the quest of finding better functionals, we consider in this paper the Perdew Burke-Ernzerhof (PBE) functional, which we believe to have the best theoretical foundation, but which leads to unacceptable errors in predicting thermochemical data (heats of formation) of molecular systems [mean absolute deviation (MAD)=16.9 kcal/mol against the extended G2 data set of 148 molecules]. Much improved thermochemistry is obtained with hybrid DFT methods that include part of the Hartree-Fock exchange [thus B3LYP (Becke's three parameter scheme combining Hartree-Fock exchange, Becke gradient corrected exchange functional and Lee-Yang Parr correlational functional) with MAD=3.1 kcal/mol and PBE0 (Perdew's hybrid scheme using PBE exchange and correlation functionals) with MAD=4.8 kcal/mol]. However we wish to continue the quest for a pure density-based DFT. Thus we optimized the four free parameters (micro, kappa, alpha, and beta) in PBE theory against experimental atomic data and the van der Waals interaction properties of Ne(2), leading to the xPBE extended functional, which significantly outperforms PBE for thermochemical properties MAD reduced to 8.0 kcal/mol while being competitive or better than PBE for predictions of geometric parameters, ionization potentials, electron affinities, and proton affinities and for the description of van der Waals and hydrogen bond interactions. Thus xPBE significantly enlarges the field of applications available for pure DFT. The functional forms thus obtained for the exchange and correlational functionals may be useful for discovering new improved functionals or formalisms. PMID- 15332953 TI - Linear response time-dependent density functional theory for van der Waals coefficients. AB - A linear response time-dependent density functional theory is described and used to calculate the dynamic polarizabilities and van der Waals C(6) coefficients of complex atom pairs. We present values of C(6) for dimers of main group atoms and the first row of transition metal atoms. PMID- 15332954 TI - Demonstration of the combination of slice imaging and Rydberg tagging for studies of photodissociation dynamics. AB - The slice-imaging variant of photofragment ion imaging is combined with Rydberg tagging. The photodissociation of NO(2) at 355 nm is used as the test system and the NO fragments are Rydberg tagged by two-photon two-color excitation via the intermediate A (2)Sigma(+) state. Images obtained by this method are compared with ion images obtained in the same apparatus using the approach of Kitsopoulos and co-workers [Rev. Sci. Instrum. 72, 3848 (2001)]. Comparable resolution and angular distributions are obtained in the two cases. It is proposed that the method demonstrated here could provide a complementary approach to existing ion imaging methods, especially where resonantly enhanced multiphoton ionization detection of fragments is problematic. PMID- 15332955 TI - Multipole-bound states of succinonitrile and other dicarbonitriles. AB - Very recently two anionic states of succinonitrile have been observed, and these two states have been interpreted as a dipole-bound state of the gauche and a quadrupole-bound state of the anti conformer. Here we study the electron binding properties of succinonitrile using high-level ab initio methods. While the dipole bound state can be investigated using well established approaches, studying the quadrupole-bound state is more challenging owing to the multiconfiguration character of its wave function. The standard methods typically applied to dipole bound anions fail, and we employ direct electron propagator based and equation-of motion coupled-cluster methods. Since there is no experience with this type of quadrupole-bound state, various basis set related and methodological aspects are examined in detail. According to our results the quadrupole moment as such plays only a minor role in binding the extra electron, whereas electron correlation effects are decisive. Our best fixed-nuclei electron binding energy is 11 meV. In view of the small binding energy the influence of the nuclear motion on the electron binding properties is examined, in particular, the torsional motion around the central carbon-carbon bond, since it is a very soft mode and the dipole and quadrupole moment depend strongly on it. Our results provide a firm basis to interpret the experimental findings and support the experimental assignments. Moreover, we discuss molecules that possess only a quadrupole-bound state, and preliminary results for dicarbonitriles of bicyclopentane and cubane are presented. PMID- 15332956 TI - Quantum and classical studies of vibrational motion of CH5+ on a global potential energy surface obtained from a novel ab initio direct dynamics approach. AB - We report a full dimensional, ab initio based potential energy surface for CH(5) (+). The ab initio electronic energies and gradients are obtained in direct dynamics calculations using second-order Moller-Plesset perturbation theory with the correlation consistent polarized valence triple zeta basis. The potential energy and the dipole moment surfaces are fit using novel procedures that ensure the full permutational symmetry of the system. The fitted potential energy surface is tested by comparing it against additional electronic energy calculations and by comparing normal mode frequencies at the three lowest-lying stationary points obtained from the fit against ab initio ones. Well-converged diffusion Monte Carlo zero-point energies, rotational constants, and projections along the CH and HH bond lengths and the tunneling coordinates are presented and compared with the corresponding harmonic oscillator and standard classical molecular dynamics ones. The delocalization of the wave function is analyzed through comparison of the CH(5) (+) distributions with those obtained when all of the hydrogen atoms are replaced by (2)H and (3)H. The classical dipole correlation function is examined as a function of the total energy. This provides a further probe of the delocalization of CH(5) (+). PMID- 15332957 TI - Calculation of the transport properties of carbon dioxide. III. Volume viscosity, depolarized Rayleigh scattering, and nuclear spin relaxation. AB - Transport properties of pure carbon dioxide have been calculated from the intermolecular potential using the classical trajectory method. Results are reported in the dilute-gas limit for volume viscosity, depolarized Rayleigh scattering, and nuclear spin relaxation for temperatures ranging from 200 to 1000 K. Three recent carbon dioxide potential energy hypersurfaces have been investigated. Calculated values for the rotational collision number for all three intermolecular surfaces are consistent with the measurements and indicate that the temperature dependence of the Brau-Jonkman correlation is not applicable for carbon dioxide. The results for the depolarized Rayleigh scattering cross section and the nuclear spin relaxation cross section show that calculated values for the generally more successful potentials differ from the observations by 9% at about 290 K, although agreement is obtained for nuclear spin relaxation at about 400 K. PMID- 15332958 TI - Theoretical studies on the electronic structure of Ti8C12 isomers. AB - Density functional calculation are performed to study Ti(8)C(12) isomers (T(d), C(3v), and D(2d)) in the neutral, cationic, and anionic charge states. C(3v) symmetry is found to be the most stable geometry for the neutral and anion, and the C(3v) and D(2d) isomers to be quasi-iso-energetic lowest for the cation. The electronic structure analysis show that d electron tends to be localized in the ground state. The theoretical assignment for the features in the experimental photoelectron spectra is given. All results obtained are in good agreement with the available experimental data and indicate that the C(3v) and D(2d) isomers may coexist in the photoelectron spectroscopy experiment. PMID- 15332959 TI - Multiconfigurational second-order perturbation study of the decomposition of the radical anion of nitromethane. AB - The doublet potential energy surfaces involved in the decomposition of the nitromethane radical anion (CH(3)NO(2) (-)) have been studied by using the multistate extension of the multiconfigurational second-order perturbation method (MS-CASPT2) in conjunction with large atomic natural orbital-type basis sets. A very low energy barrier is found for the decomposition reaction: CH(3)NO(2) (-)- >[CH(3)NO(2)](-)-->CH(3)+NO(2) (-). No evidence has been obtained on the existence of an isomerization channel leading to the initial formation of the methylnitrite anion (CH(3)ONO(-)) which, in a subsequent reaction, would yield nitric oxide (NO). In contrast, it is suggested that NO is formed through the bimolecular reaction: CH(3)+NO(2) (-)-->[CH(3)O-N-O](-)-->CH(3)O(-)+NO. In particular, the CASSCF/MS-CASPT2 results indicate that the methylnitrite radical anion CH(3)ONO(-) does not represent a minimum energy structure, as concluded by using density functional theory (DFT) methodologies. The inverse symmetry breaking effect present in DFT is demonstrated to be responsible for such erroneous prediction. PMID- 15332960 TI - Intense-field modulation of NO2 multiphoton dissociation dynamics. AB - We report on the dynamics of multiphoton excitation and dissociation of NO(2) at wavelengths between 395 and 420 nm and intensities between 4 and 10 TW cm(-2). The breakup of the molecule is monitored by NO A (2)Sigma(+)n(')=1,0-->X (2)Pi(r)n(")=0 fluorescence as a function of time delay between the driving field and a probe field which depletes the emission. It is found that generation of n(')=0 and 1 NO A (2)Sigma(+) results in different fluorescence modulation patterns due to the intense probe field. The dissociation dynamics are interpreted in terms of nuclear motions over light-induced potentials formed by coupling of NO(2) valence and Rydberg states to the applied field. Based on this model, it is argued that the time and intensity dependences of A (2)Sigma(+)n(')=0-->X (2)Pi(r)n(")=0 fluorescence are consistent with delayed generation of NO A (2)Sigma(+)n(')=0 via a light-induced bond-hardening brought about by the transient coupling of the dressed A (2)B(2) and Rydberg 3ssigma (2)Sigma(g) (+) states of the parent molecule. The increasingly prompt decay of A (2)Sigma(+)n(')=1-->X (2)Pi(r)n(")=0 fluorescence with increasing intensity, on the other hand, is consistent with a direct surface crossing between the X (2)A(1) and 3ssigma (2)Sigma(g) (+) dressed states to generate vibrationally excited products. PMID- 15332961 TI - Electron propagator theory calculations of molecular photoionization cross sections: the first-row hydrides. AB - Together with ionization potentials, cross sections provide valuable information for the interpretation of photoelectron spectra. We have developed a program to perform ab initio calculations of photoionization cross sections within the electric dipole approximation using electron propagator theory. Applications to the first-row hydrides CH(4), NH(3), H(2)O, and HF, using several approximations for the propagator self-energy and the plane-wave and orthogonalized-plane-wave approximations to represent the photoelectron, as well as comparison to experimental data, are presented. This program is implemented within the quantum chemistry package GAUSSIAN. PMID- 15332962 TI - An ab initio potential energy surface and vibrational states of MgH2(1(1)A'). AB - A three-dimensional global potential energy surface for the ground electronic state of MgH(2) is constructed from more than 3000 ab initio points calculated using the internally contracted multireference configuration interaction method with the Davidson correction at the complete basis set limit. Low-lying vibrational energy levels of MgH(2) and MgD(2) are calculated using the Lanczos algorithm, and found to be in good agreement with known experimental band origins. The majority of the vibrational energy levels up to 8000 cm(-1) are assigned with normal mode quantum numbers. However, our results indicate a gradual transition from a normal mode regime for the stretching vibrations at low energies to a local mode regime near 7400 cm(-1), as evidenced by a decreasing energy gap between the (n(1),0,0) and (n(1)-1,0,1) vibrational states and bifurcation of the corresponding wave functions. PMID- 15332963 TI - New electronic spectra of the HCCl and DCCl A-X vibronic bands. AB - The dispersed fluorescence spectra following the excitation of several A<--X vibronic bands of HCCl and DCCl at visible wavelengths were successfully acquired in a discharge supersonic free jet expansion using an intensified charge-coupled device detector. The dispersed fluorescence spectra reveal more details of the X(1) A(') state vibrational structure in these molecules than previous reports. Dispersed fluorescence spectra of all four isotopomers (HC(35)Cl, HC(37)Cl, DC(35)Cl, and DC(37)Cl) were obtained. These dispersed fluorescence spectra exhibit the vibrational structures up to approximately 6000 cm(-1) above the zero point level and determine the vibrational structures of HC(37)Cl and DC(37)Cl. Complete vibrational parameters including fundamental frequencies, anharmonicities, and coupling constants were determined for the HCCl/DCCl X(1) A(') state. Furthermore, perturbations from the background triplet state a(3) A(") and emission to triplet state levels were observed in the spectra. The singlet-triplet energy gap from the zero-point level could be determined to be 2167 cm(-1) (6.20+/-0.05 kcal/mol) in HCCl and to be 2187 cm(-1) (6.25+/-0.05 kcal/mol) in DCCl. Additionally, some of the A<--X excitation spectrum are reported for HCCl and DCCl. PMID- 15332964 TI - Quadrupole and octupole polarizabilities for the ground states of lithiumlike systems from Z=3 to 20. AB - The quadrupole and octupole polarizabilities for the ground states of lithiumlike systems from Z=3 to 20 are calculated with the full-core plus correlation method. For the neutral lithium atom, the typical patterns of convergence of the quadrupole and octupole polarizabilities are analyzed. The calculated quadrupole and octupole polarizabilities of the ground state for lithium atom are compared with the previous theoretical results obtained by other methods; our predictions agree with the most accurate reports in the literature very well. For lithiumlike ions, our prediction may provide valuable reference data for other accurate theoretical calculations in future. PMID- 15332965 TI - Hydrogen loss from nucleobase nitrogens upon electron attachment to isolated DNA and RNA nucleotide anions. AB - Electron transfer to isolated nucleotide monoanions in collisions with Na vapor induces hydrogen loss from nitrogen of the transient nucleobase anion. The cross section for this process is linearly correlated with the number of N-H hydrogens and is highest for guanine. The process is much faster than microseconds since only dehydrogenated dianions survived for mass spectrometric detection. The lifetime of the adenosine 5(')-monophospate dianions was measured to be 0.2 ms in an electrostatic ion storage ring but also a longer-lived component with a lifetime of at least 10 ms was identified. Implications of dissociation along the N-H coordinate for a nucleotide in DNA are briefly discussed in terms of Watson Crick base pairs. PMID- 15332966 TI - Interaction potentials and rovibrational spectroscopy of He(N)-OCS complexes. AB - We present a new vibrationally averaged He-OCS potential energy surface that is obtained from a combination of Moller-Plesset perturbation theory for the helium molecule interaction and coupled cluster theory for the intramolecular vibrational potential. Employing this potential in quantum Monte Carlo calculations for He(N)-OCS complexes shows a blueshift of the OCS vibration for small N that is followed by a transition to a redshift for larger N. The size dependence of the vibrational shift is in good agreement with recent experimental measurements. We then make a comparative study of the effective rotational spectroscopic constants B(eff) and D(eff) calculated for small N values with this vibrationally averaged potential, with the corresponding values obtained from three previous He-OCS potentials. We find that the vibrationally averaged potential provides the most accurate description of the spectroscopic constants over the size range N=1-8 for which experimental data are available. We rationalize this improved description in terms of the detailed differences in the secondary minimum and saddle point regions of the underlying He-OCS interaction potential, in addition to the behavior at the lowest potential minimum. This analysis indicates that the spectroscopy of complexes with N>1 provides valuable information on the shape of the potential energy surface in regions that are not accessed by the N=1 He-OCS complex, but that are important for understanding the molecular spectroscopy in larger complexes and in droplets. PMID- 15332967 TI - Low-temperature nucleation in a kinetic Ising model under different stochastic dynamics with local energy barriers. AB - Using both analytical and simulational methods, we study low-temperature nucleation rates in kinetic Ising lattice-gas models that evolve under two different Arrhenius dynamics that interpose between the Ising states a transition state representing a local energy barrier. The two dynamics are the transition state approximation [T. Ala-Nissila, J. Kjoll, and S. C. Ying, Phys. Rev. B 46, 846 (1992)] and the one-step dynamic [H. C. Kang and W. H. Weinberg, J. Chem. Phys. 90, 2824 (1989)]. Even though they both obey detailed balance and are here applied to a situation that does not conserve the order parameter, we find significant differences between the nucleation rates observed with the two dynamics, and between them and the standard Glauber dynamic [R. J. Glauber, J. Math. Phys. 4, 294 (1963)], which does not contain transition states. Our results show that great care must be exercised when devising kinetic Monte Carlo transition rates for specific physical or chemical systems. PMID- 15332968 TI - The behavior of fluids near solutes: a density functional theory and computer simulation study. AB - The density distribution of solvent near a solute particle is studied using density functional theory and Monte Carlo simulation. The fluid atoms interact with each other via a hard sphere plus Yukawa potential, and interact with the solute via a hard sphere potential. For small solute sizes, the solvent displays liquidlike ordering near the particle. When the solute become larger, a drying transition is observed at state points near the coexistence conditions of the solvent. These predictions are similar to those of a recent theory for the hydrophobic effect by Lum, Chandler, and Weeks [J. Phys. Chem. 103, 4570 (1999)], although a comparison with simulations shows that the theory of this work is quantitatively more accurate. The connection between density functional methods and the LCW approach is also established. PMID- 15332969 TI - Density functional theory for semiflexible and cyclic polyatomic fluids. AB - The effects of bond angle and chain stiffness on the structures of semiflexible polyatomic fluids are investigated by incorporating the bending potential into a density functional theory [Y. X. Yu and J. Z. Wu, J. Chem. Phys. 117, 2368 (2002)] that combines a modified fundamental measure theory for the excluded volume effects and the first-order thermodynamics perturbation theory for the chain connectivity. The refined density functional theory faithfully reproduces the density profiles and conformational properties of a variety of triatomic fluids near a hard wall in which extensive Monte Carlo simulation data are available. In particular, the theory is able to capture the structures of rigid cyclic trimers where all segments are identical. The variation of local density profiles with respect to the chain length of confined polyatomic fluids is also explored. For quadratomic fluids confined in slit pores, the density profile of the middle segments exhibits novel double peaks that are absent in a fully flexible chain model. In addition, the density functional theory is applied to predicting the conformational properties and adsorption behavior of heterogeneous triatomic fluids of type "ABB" mimicking surfactant molecules. The competition between surface adsorption and self-association of trimers consisting of surface active and self-binding "A" segments and neutral "B" segment is explored. PMID- 15332970 TI - Phase-stabilized two-dimensional electronic spectroscopy. AB - Two-dimensional (2D) spectroscopy is a powerful technique to study nuclear and electronic correlations between different transitions or initial and final states. Here we describe in detail our development of inherently phase-stabilized 2D Fourier-transform spectroscopy for electronic transitions. A diffractive-optic setup is used to realize heterodyne-detected femtosecond four-wave mixing in a phase-matched box geometry. Wavelength tunability in the visible range is accomplished by means of a 3 kHz repetition-rate laser system and optical parametric amplification. Nonlinear signals are fully characterized by spectral interferometry. Starting from fundamental principles, we discuss the origin of phase stability and the precise calibration of excitation-pulse time delays using movable glass wedges. Automated subtraction of undesired scattering terms removes experimental artifacts. On the theoretical side, the response-function formalism is extended to describe molecules with three electronic levels, and the shape of 2D spectral features is discussed. As an example for this technique, experimental 2D spectra are shown for the dye molecule Nile Blue in acetonitrile at 595 nm, recorded for a series of population times. Simulations explore the influence of different model parameters and qualitatively reproduce the experimental results. We show that correlations between different electronically excited states can be determined from the spectra. The technique described here can be used to measure the third-order response function of complex systems covering several electronic transitions. PMID- 15332971 TI - On the mechanism of reorientational and structural relaxation in supercooled liquids: the role of border dynamics and cooperativity. AB - Molecular dynamics simulation and analysis based upon the many-body potential energy landscape (PEL) are employed to characterize single molecule reorientation and structural relaxation, and their interrelation, in deeply supercooled liquid CS(2). The rotational mechanism changes from small-step Debye diffusion to sudden large angle reorientation (SLAR) as the temperature falls below the mode-coupling temperature T(c). The onset of SLAR is explained in terms of the PEL; it is an essential feature of low-T rotational dynamics, along with the related phenomena of dynamic heterogeneity and the bifurcation of slow and fast relaxation processes. A long trajectory in which the system is initially trapped in a low energy local minimum, and eventually escapes, is followed in detail, both on the PEL and in real space. During the trapped period, "return" dynamics occurs, always leading back to the trap. Structural relaxation is identified with irreversible escape to a new trap. These processes lead to weak and strong SLAR, respectively; strong SLAR is a clear signal of structural relaxation. Return dynamics involves small groups of two to four molecules, while a string-like structure composed of all the active groups participates in the escape. It is proposed that, rather than simple, nearly instantaneous, one-dimensional barrier crossings, relaxation involves activation of the system to the complex, multidimensional region on the borders of the basins of attraction of the minima for an extended period. PMID- 15332972 TI - Dynamical density functional theory and its application to spinodal decomposition. AB - We present an alternative derivation of the dynamical density functional theory for the one-body density profile of a classical fluid developed by Marconi and Tarazona [J. Chem. Phys. 110, 8032 (1999)]. Our derivation elucidates further some of the physical assumptions inherent in the theory and shows that it is not restricted to fluids composed of particles interacting solely via pair potentials; rather it applies to general, multibody interactions. The starting point for our derivation is the Smoluchowski equation and the theory is therefore one for Brownian particles and as such is applicable to colloidal fluids. In the second part of this paper we use the dynamical density functional theory to derive a theory for spinodal decomposition that is applicable at both early and intermediate times. For early stages of spinodal decomposition our nonlinear theory is equivalent to the (generalized) linear Cahn-Hilliard theory, but for later times it incorporates coupling between different Fourier components of the density fluctuations (modes) and therefore goes beyond Cahn-Hilliard theory. We describe the results of calculations for a model (Yukawa) fluid which show that the coupling leads to the growth of a second maximum in the density fluctuations, at a wave number larger than that of the main peak. PMID- 15332973 TI - Shape and motility of a model cell: a computational study. AB - We have investigated the shape, size, and motility of a minimal model of an adherent biological cell using the Monte Carlo method. The cell is modeled as a two dimensional ring polymer on the square lattice enclosing continuously polymerizing and depolymerizing actin networks. Our lattice model is an approximate representation of a real cell at a resolution of one actin molecule, 5 nm. The polymerization kinetics for the actin network are controlled by appropriate reaction probabilities which correspond to the correct experimental reaction rates. Using the simulation data we establish various scaling laws relating the size of the model cell to the concentration of polymerized and unpolymerized actin molecules and the length of the enclosing membrane. The computed drift velocities, which characterize the motility of the cell, exhibit a maximum at a certain fraction of polymerized actin which agrees with physiological fractions observed in experiments. The appearance of the maximum is related to the competition between the polymerization-induced protrusion of the membrane and the concomitant suppression of membrane fluctuations. PMID- 15332974 TI - Na-montmorillonite hydrates under ethane rich reservoirs: NPzzT and muPzzT simulations. AB - Na-montmorillonite hydrates in presence of ethane molecules are studied by means of hybrid Monte Carlo simulations in the NP(zz)T and muP(zz)T ensembles. The NP(zz)T ensemble allows us to study the interlaminar distance as a function of water and ethane content. These data show clear plateaus for lower ethane contents and mainly for water contents consistent with the formation of a single water layer. In addition, from this ensemble the structure for some of these interlaminar compositions were analyzed. For systems containing few ethane molecules and water enough to complete a single layer, it was observed that ethane mainly situates close to the interlayer midplane and adopts a nearly parallel arrangement to the clay surface. On the other hand, the muP(zz)T ensemble allows us to determine the interlaminar distance and water-ethane content for any specific reservoir. Here, some important findings are the following: the partial exchange of water by ethane molecules that enhances for decreasing the water vapor pressure; the obtention of a practically constant interlaminar space distance as a function of the water vapor pressure; the conservation of ion solvation shells; the enhancement of the water-ethane exchange for burial conditions; and finally, the incapability for a dehydrated clay mineral to swell in a dry and rich ethane atmosphere. PMID- 15332975 TI - A model for the phase stability of arbitrary nanoparticles as a function of size and shape. AB - A thermodynamic model describing relative stability of different shapes for nanoparticles as a function of their size was developed for arbitrary crystalline solids and applied to group IV semiconductors. The model makes use of various surface, edge and corner energies, and takes into account surface tension. Approximations and importance of each term of the model were analyzed. The predictions for clean and hydrogenated diamond nanoparticles are compared to explicitly calculated density functional results. It is shown that diamond nanocrystal morphology is markedly different from silicon and germanium. PMID- 15332976 TI - Low-energy electron scattering cross section for the production of CO within solid films of carbon dioxide. AB - We report absolute electron scattering cross sections sigma(p) for the production of CO within thin solid film of carbon dioxide (CO(2)) condensed on a solid Ar substrate. The CO fragments, which remain trapped within the bulk of the carbon dioxide film, are detected in situ by recording energy losses to their lowest triplet electronic state a (3)Pi using high-resolution electron-energy-loss spectroscopy. The production of CO is studied as a function of the electron exposure, film thickness, and incident electron energy between 2 and 30 eV, a range within which most of the secondary electrons are created in systems irradiated by high-energy particles. The energy dependence is characterized by a feature around 4 eV with sigma(p)=(7.0+/-4.0)x10(-18) cm(2), a minimum around 7 eV, a strong rise up to a large and broad maximum around 15 eV with sigma(p)=(5.4+/-2.5)x10(-17) cm(2), a decrease to a minimum around 18.5 eV, and finally a monotonous increase up to 30 eV. The CO production is discussed in terms of the formation of electron resonances or transient anion states, which may lead directly to the fragmentation of the molecule via dissociative electron attachment or indirectly by decaying into an entirely repulsive part of the corresponding excited neutral and positive ion states. PMID- 15332977 TI - Molecular dynamics simulation of optically trapped colloidal particles at an oil water interface. AB - Using molecular dynamics simulations, we calculate the net force on a colloidal particle trapped by an optical tweezer and confined within a particle monolayer which is in motion relative to the trapped particle. The calculations are compared with recent experimental data on polystyrene particles located at an oil water interface. Good agreement between theory and experiment is obtained over the investigated range of lattice constants for an interaction mechanism between the polystyrene particles which is dominated by an effective dipole-dipole potential. The assumed interaction mechanism is consistent with the formation of surface charge dipoles at the particle-oil interface due to the dissociaton of the hydrophilic sulfate headgroups at the surface of the polystyrene particles. A possible physical mechanism for the formation of the surface charge dipoles, involving a diffuse cloud of fully hydrated counterions, is described, and the fraction of surface groups contributing to the formation of surface charge dipoles is estimated to be of the order of 10(-1) for the present system. PMID- 15332978 TI - A vibronic approach to the band-filling and temperature-dependent metal-insulator transition. AB - A model of vibronic origin is used to investigate the important issue of metal insulator transition in low-dimensional materials. For zero temperature, the stability of the single-band model chain is controlled by the competition between the internal electron-phonon coupling and the nearest-neighbor hopping integral. Assuming one particular deformation mode, one can analytically derive an instability criterion in which the band filling is explicitly included. The carrier doping directly controls the stability of a one-dimensional chain. For a half-filled band, the Peierls instability is recovered. For finite temperatures, a similar criterion is derived and can be used to investigate the metal-insulator transition temperatures. PMID- 15332979 TI - Ionic effect on combing of single DNA molecules and observation of their force induced melting by fluorescence microscopy. AB - Molecular combing is a powerful and simple method for aligning DNA molecules onto a surface. Using this technique combined with fluorescence microscopy, we observed that the length of lambda-DNA molecules was extended to about 1.6 times their contour length (unextended length, 16.2 microm) by the combing method on hydrophobic polymethylmetacrylate coated surfaces. The effects of sodium and magnesium ions and pH of the DNA solution were investigated. Interestingly, we observed force-induced melting of single DNA molecules. PMID- 15332980 TI - Local-field effects on the spontaneous emission rate of CdTe and CdSe quantum dots in dielectric media. AB - The refractive index dependence of the spontaneous emission rate is determined using organically capped CdSe and CdTe quantum dots as probes. The radiative lifetime of the exciton emission is measured in a variety of apolar solvents with refractive indices n between 1.37 and 1.50. It is demonstrated that quantum dots provide a model system for testing theories on the influence of local-field effects on the spontaneous emission rate. The experimentally observed influence of n on the radiative lifetime is smaller than predicted by well-known models for local-field corrections but is in good agreement with a recently developed fully microscopic model for the local-field enhancement of the spontaneous emission rate. PMID- 15332981 TI - Field theoretical approach to electrochemical deposition. AB - In this work we present an application of the lambdaphi(4) field theoretical model to the adsorption of atoms and molecules on metallic surfaces-the electrochemical deposition. The usual approach to this system consists in the computational simulation using Monte Carlo techniques of an effective lattice-gas Hamiltonian. We construct an effective model towards a comparison between the lattice-gas Hamiltonian and the discrete version of the lambdaphi(4) Hamiltonian, obtaining the relationships between the model parameters and electrochemical quantities. The lambdaphi(4) model is studied in the mean field approximation, and the results are fitted and compared to numerical simulated and experimental data. PMID- 15332982 TI - Large-scale molecular dynamics simulations of alkanethiol self-assembled monolayers. AB - Large-scale molecular dynamics simulations of self-assembled alkanethiol monolayer systems have been carried out using an all-atom model involving a million atoms to investigate their structural properties as a function of temperature, lattice spacing, and molecular chain length. Our simulations show that the alkanethiol chains of 13-carbons tilt from the surface normal by a collective angle of 25 degrees along next-nearest-neighbor direction at 300 K. The tilt structure of 13-carbon alkanethiol system is found to depend strongly on temperature and exhibits hysteresis. At 350 K the 13-carbon alkanethiol system transforms to a disordered phase characterized by small collective tilt angle, flexible tilt direction, and random distribution of backbone planes. The tilt structure also depends on lattice spacing: With increasing lattice spacing a the tilt angle increases rapidly from a nearly zero value at a=4.7 A to as high as 34 degrees at a=5.3 A at 300 K for 13-carbon alkanethiol system. Finally, the effects of the molecular chain length on the tilt structure are significant at high temperatures. PMID- 15332983 TI - Ammonia adsorption on and diffusion into thin ice films grown on Pt(111). AB - Ammonia adsorption on and diffusion into thin ice films grown on a Pt(111) surface were studied using Fourier transform infrared spectroscopy (FTIR) and thermal desorption spectroscopy. After exposing the crystalline ice film to ammonia molecules at 45 K (ammonia/ice film), we have detected an intriguing feature at 1470 cm(-1) in the FTIR spectra, which is derived from the adsorption of ammonia on the ice with a characteristic structure which appears in thin film range. The peak intensity of this feature decreases gradually as the thickness of the substrate ice increases. In addition, we have detected a feature at 1260 cm( 1) which appears after annealing the ammonia/ice film. The feature corresponds to the ammonia molecules which reach the ice/Pt(111) interface through the ice film. Intriguingly, the intensity of this feature decreases with the ice thickness and there is a linear relation of the peak intensity of the features at 1470 and 1260 cm(-1). We propose a model in which the solubility of the ammonia molecules is much higher for the thin ice film than that for the ideal ice. PMID- 15332984 TI - Evidence from first principles calculations for a bent CO2 intermediate in the oxidation of carbon monoxide on the Cu (110) surface. AB - We have carried out first principles plane wave density-functional theory calculations to study the adsorption of CO molecule on a clean and unreconstructed Cu (110) surface at 1/12 monolayer coverage and have investigated the subsequent oxidation by preadsorbed oxygen atoms. As found experimentally, the CO adsorbs perpendicular to the surface plane through the carbon atom; the top site was found to be the most favorable position for CO adsorption although the short-bridge site is only slightly less stable. Surprisingly, for a sparely oxidized surface with O atoms adsorbed in hollow sites the coadsorption energy is slightly negative for only the above two CO sites which have therefore been used as starting points to explore the energy surface of the oxidation reaction. We have confirmed the existence of bent CO(2) surface intermediate as previously suggested from experimental studies. Using the nudged elastic band method, we have characterized a two step reaction which involves the formation of this intermediate. The results suggest that the rate determining step of the oxidation reaction is the formation of the intermediate and the energy barrier (200 meV) is close to although smaller than experimentally estimated values. PMID- 15332985 TI - Solvent-driven dewetting and rim instability. AB - An experimental method suitable for reproducible results has been used to investigate dewetting behavior of thin films of solvent-laden polymer. This solvent-driven dewetting enables one to change spreading coefficient by an order of magnitude that is not readily realizable in thermal dewetting and to study polar interactions that have not been fully exploited experimentally. While the film instability is similar to that found in thermal dewetting, the rim instability is quite different. Two different types of the rim instability have been found. With a polar solvent, the rim instability changes from one type to another with increasing film thickness whereas the unstable rim becomes stable for an apolar solvent. PMID- 15332986 TI - Lattice-gas modeling of CO adlayers on Pd(100). AB - Using a lattice-gas model with pairwise interactions, we study the ordered structures, coverage dependence of the heat of adsorption, and other experimentally observable behavior of adsorbed CO overlayers on Pd(100) single crystal surfaces. Transfer matrix and Monte Carlo methods give accurate information regarding the lattice-gas model that often contradicts simple mean field-like analysis. We demonstrate the usefulness of the model by reproducing experimental results over a large range of pressures and temperatures. PMID- 15332987 TI - Disjoining pressure measurements using a microfabricated groove for a molecularly thin polymer liquid film on a solid surface. AB - A method for measuring disjoining pressure of a molecularly thin liquid film on a solid surface by using a microfabricated groove has been developed. The shape of the meniscus of a thin film in the microgroove was measured with an atomic force microscope, and the disjoining pressure was obtained from the capillary pressure obtained from the measured curvature of the meniscus. Our method is applicable to a film with a thickness greater than the diameter of gyration in the polymer molecule. Moreover, the method can detect the changes in the disjoining pressure caused by ultraviolet light irradiation, and it is effective in investigating the intermolecular interaction between a thin film and a solid surface. PMID- 15332988 TI - Cranklike conformational transitions in polyethylene. AB - Molecular dynamics simulations of a variety of polymeric systems provide the evidence for two different kinds of conformational transitions: independent single bond transitions and cranklike transitions (or correlated bond transitions). While single bond transitions can be rationalized according to standard theories of activated processes controlled by the saddle point crossing, a more complex description is required for the other type of transitions. In a recent work devoted to the analysis of the simplified chain model with three rotors [B. Nigro and G. J. Moro, J. Phys. Chem. B 106, 7365 (2002)], a theory has been proposed for cranklike transitions represented as a kinetic process between equilibrium states differing by two torsional angles (i.e., two bond transitions). Moreover their rate coefficients were estimated on the basis of a local expansion about the bifurcation of the separatrices departing from the potential function maximum. In the present work the same theory is applied to a model for long alkyl chains in solution, in order to rationalize the behavior of cranklike transitions in polyethylene and to recognize the molecular features controlling them. We obtain probabilities of occurrence of cranklike transitions in substantial agreement with simulation results. Furthermore, the theory is capable of explaining the dependence of the rate on the separation between the two reactive bonds, as well as the dependence on the conformational state of the starting configuration. In particular, selection rules for next-to-nearest neighbor transitions are recovered from the shape of the separatrices and the location of the corresponding bifurcations. PMID- 15332989 TI - Double proton transfer in the isolated and DNA-embedded guanine-cytosine base pair. AB - The energetics and dynamics of double proton transfer (DPT) is investigated theoretically for the Watson-Crick conformation of the guanine-cytosine (GC) base pair. Using semiempirical density functional theory the isolated and DNA-embedded GC pair is considered. Differences in the energetics and dynamics of DPT thus addresses the question of how relevant studies of isolated base pairs are for the understanding of processes occurring in DNA. Two-dimensional potential energy surfaces involving the transferring hydrogen atoms and the proton donors and acceptors are presented for both systems. The DPT reaction is accompanied by a contraction of the distance between the two bases with virtually identical energetic barriers being 18.8 and 18.7 kcal/mol for the isolated and DNA-embedded system, respectively. However, the transition state for DPT in the DNA-embedded GC pair is offset by 0.1 A to larger N-H separation compared to the isolated GC pair. Using activated ab initio molecular dynamics, DPT is readily observed for the isolated base pair with a minimal amount of 21.4 kcal/mol of initial average kinetic energy along the DPT normal mode vector. On a time scale of approximately 100 fs DPT has occurred and the excess energy is redistributed. For the DNA embedded GC pair considerably more kinetic energy is required (30.0 kcal/mol) for DPT and the process is completed within one hydrogen vibration. The relevance of studies of isolated base pairs and base pair analogs in regard of reactions or properties involving DNA is discussed. PMID- 15332990 TI - Geometry, dipole moment, polarizability and first hyperpolarizability of polymethineimine: an assessment of electron correlation contributions. AB - We have investigated the geometries as well as the longitudinal dipole moment (micro), polarizability (alpha), and first hyperpolarizability (beta) of polymethineimine oligomers using different approaches [Hartree-Fock (HF), second order Moller-Plesset (MP2), and hybrid density functional theory (DFT) methods (B3LYP and PBE0)] for evaluating the geometries and the nonlinear optical properties. It turns out that (i) HF and the selected DFT methods provide the incorrect sign for beta of short and medium size oligomers. (ii) The B3LYP and PBE0 electron correlation correction are too small for micro, too large for alpha, and for some oligomer lengths, they are in the wrong direction for beta. (iii) On the contrary to polyacetylene, the hybrid-DFT geometries are in poor agreement with MP2 geometries; the former showing much smaller bond length alternations. PMID- 15332991 TI - Field-induced alignment of a smectic-A phase: a time-resolved x-ray diffraction investigation. AB - The field-induced alignment of a smectic-A phase is, in principle, a complicated process involving the director rotation via the interaction with the field and the layer rotation via the molecular interactions. Time-resolved nuclear magnetic resonance spectroscopy has revealed this complexity in the case of the director alignment, but provides no direct information on the motion of the layers. Here we describe a time-resolved x-ray diffraction experiment using synchrotron radiation to solve the challenging problem of capturing the diffraction pattern on a time scale which is fast in comparison with that for the alignment of the smectic layers. We have investigated the alignment of the smectic-A phase of 4 octyl-4(')-cyanobiphenyl by a magnetic field. The experiment consists of creating a monodomain sample of the smectic-A phase by slow cooling from the nematic phase in a magnetic field with a flux density of 7 T. The sample is then turned quickly through an angle phi(0) about an axis parallel to the x-ray beam direction but orthogonal to the field. A sequence of two-dimensional small angle x-ray diffraction patterns are then collected at short time intervals. Experiments were carried out for different values of phi(0), and at different temperatures. The results show that the alignment behavior changes fundamentally when phi(0) exceeds 45 degrees, and that there is a sharp change in the alignment process when the temperature is less than 3 degrees C below the smectic-A-nematic transition. The results of the x-ray experiments are in broad agreement with the NMR results, but reveal major phenomena concerning the maintenance of the integrity of the smectic-A layer structure during the alignment process. PMID- 15332992 TI - Capillary bridging and long-range attractive forces in a mean-field approach. AB - When a mixture is confined, one of the phases can condense out. This condensate, which is otherwise metastable in the bulk, is stabilized by the presence of surfaces. In a sphere-plane geometry, routinely used in atomic force microscope and surface force apparatus, it can form a bridge connecting the surfaces. The pressure drop in the bridge gives rise to additional long-range attractive forces between them. By minimizing the free energy of a binary mixture we obtain the force-distance curves as well as the structural phase diagram of the configuration with the bridge. Numerical results predict a discontinuous transition between the states with and without the bridge and linear force distance curves with hysteresis. We also show that similar phenomenon can be observed in a number of different systems, e.g., liquid crystals and polymer mixtures. PMID- 15332993 TI - Counterion associative behavior with flexible polyelectrolytes. AB - At low ionic strength, organic counterions dress a flexible charged polymer as measured directly by small-angle neutron scattering and neutron spin-echo spectroscopy. This dressed state, quantified by the concentration dependence of the static correlation length, illustrates the polymer-counterion coupled nature on the nanometer length scale. The counterions, made visible by selective hydrogen and deuterium labeling, undress from the polymeric template by addition of sodium chloride. The addition of this electrolyte leads to two effects: increased Debye electrostatic screening and decoupled organic counterion-polymer correlations. Neutron spin-echo spectroscopy measures a slowing down of the effective diffusion coefficient of the labeled counterions at the length scale of 8 nm, the static correlation length, indicating the nanosecond counterion dynamics mimics the polymer. These experiments, performed with semidilute solutions of tetramethylammonium poly(styrene sulfonate) [(h-TMA(+)) d-PSS], apply to relevant biopolymers including single and double stranded DNA and unfolded proteins, which undergo orchestrated dynamics of counterions and chain segments to fold, unfold, and assemble. PMID- 15332994 TI - Nematic-amorphous polymer interfaces in the presence of a compatibilizer. AB - We introduce and apply a variant of a dynamic self-consistent field simulation in two dimensions to predict the structure of interfaces between a nematic and an amorphous polymer compatibilized by a diblock copolymer. First, we investigate the effect of the nematic order on the polymer polymer interface without compatibilizer. Then we include the compatibilizer and consider two interfacial setups previously used in experiments, i.e., the bilayer setup and the trilayer setup. In the bilayer setup the diblock copolymer is mixed into the amorphous homopolymer and migrates to the interface in the course of the simulation forming a layered structure. We compare the amount of copolymer at the interface for initial concentrations of the copolymer below and above the critical micelle concentration. In the trilayer setup the initial thickness of the diblock copolymer is varied. The resulting interfacial morphology evolves in the competition between the lamellar structure induced by the interface and a micellar structure, which is intrinsic to the copolymer. PMID- 15332996 TI - Nuclear translocation of cleaved LyGDI dissociated from Rho and Rac during Trp53 dependent ionizing radiation-induced apoptosis of thymus cells in vitro. AB - LyGDI inhibits the dissociation of GDP from Rho family GTPases and is found in abundance in hematopoietic cells. Here we report truncation of LyGDI after irradiation in mouse 3SB thymus cells. A 21-kDa fragment of LyGDI, resulting from activated caspase 3-induced cleavage at an N-terminal consensus site following the Asp(18) residue, accumulated at peak quantities between 5 and 12 h after irradiation. Cleavage of LyGDI was inhibited by the caspase inhibitor benzoyloxycarbonyl-Val-Asp-fluoromethylketone. Subcellular fractionation and immunofluorescence revealed the truncated 21-kDa fragment of LyGDI within the nuclear fraction of irradiated 3SB cells, whereas full-length LyGDI was found only in the cytoplasmic fraction. Truncated LyGDI within the nucleus had no association with the Rho family proteins RhoA and Rac1, since these proteins were observed only in the cytoplasmic fractions. These data demonstrate that regulation of Rho family GTPases by LyGDI is disrupted during apoptosis, suggesting that fragmentation of LyGDI implicates the transmission of a signal from the cytoplasm to the nucleus during Trp53-dependent apoptosis of thymus cells after irradiation. PMID- 15332997 TI - Computerized video time lapse study of cell cycle delay and arrest, mitotic catastrophe, apoptosis and clonogenic survival in irradiated 14-3-3sigma and CDKN1A (p21) knockout cell lines. AB - Computerized video time lapse (CVTL) microscopy was used to observe cellular events induced by ionizing radiation (10-12 Gy) in nonclonogenic cells of the wild-type HCT116 colorectal carcinoma cell line and its three isogenic derivative lines in which p21 (CDKN1A), 14-3-3sigma or both checkpoint genes (double knockout) had been knocked out. Cells that fused after mitosis or failed to complete mitosis were classified together as cells that underwent mitotic catastrophe. Seventeen percent of the wild-type cells and 34-47% of the knockout cells underwent mitotic catastrophe to enter generation 1 with a 4N content of DNA, i.e., the same DNA content as irradiated cells arrested in G(2) at the end of generation 0. Radiation caused a transient division delay in generation 0 before the cells divided or underwent mitotic catastrophe. Compared with the division delay for wild-type cells that express CDKN1A and 14-3-3sigma, knocking out CDKN1A reduced the delay the most for cells irradiated in G(1) (from approximately 15 h to approximately 3- 5 h), while knocking out 14-3-3sigma reduced the delay the most for cells irradiated in late S and G(2) (from approximately 18 h to approximately 3-4 h). However, 27% of wild-type cells and 17% of 14-3-3sigma(-/-) cells were arrested at 96 h in generation 0 compared with less than 1% for CDKN1A(-/-) and double-knockout cells. Thus expression of CDKN1A is necessary for the prolonged delay or arrest in generation 0. Furthermore, CDKN1A plays a crucial role in generation 1, greatly inhibiting progression into subsequent generations of both diploid cells and polyploid cells produced by mitotic catastrophe. Thus, in CDKN1A-deficient cell lines, a series of mitotic catastrophe events occurred to produce highly polyploid progeny during generations 3 and 4. Most importantly, the polyploid progeny produced by mitotic catastrophe events did not die sooner than the progeny of dividing cells. Death was identified as loss of cell movement, i.e. metabolic activity. Thus mitotic catastrophe itself is not a direct mode of death. Instead, apoptosis during interphase of both uninucleated and polyploid cells was the primary mode of death observed in the four cell types. Knocking out either CDKN1A or 14-3-3sigma increased the amount of cell death at 96 h, from 52% to approximately 70%, with an even greater increase to 90% when both genes were knocked out. Thus, in addition to effects of CDKN1A and 14-3-3sigma expression on transient cell cycle delay, CDKN1A has both an anti-proliferative and anti-apoptosis function, while 14-3-3sigma has only an anti-apoptosis function. Finally, the large alterations in the amounts of cell death did not correlate overall with the small alterations in clonogenic survival (dose-modifying ratios of 1.05-1.13); however, knocking out CDKN1A resulted in a decrease in arrested cells and an increase in survival, while knocking out 14-3-3sigma resulted in an increase in apoptosis and a decrease in survival. PMID- 15332998 TI - Effect of medium on chromatin damage in bystander mammalian cells. AB - In the present study, we examined the potential contribution of irradiated medium to the bystander effect using custom-made double-Mylar stainless steel rings. Exponentially growing human-hamster hybrid (A(L)) cells were plated on either one or both sides of double-Mylar dishes 2-4 days before irradiation. One side (with or without cells) was irradiated with alpha particles using the track segment mode of a 4 MeV Van de Graaff accelerator at the Radiological Research Accelerator Facility of Columbia University. Since alpha particles can traverse only a very limited distance (around 23 microm in water), cells plated on the other side of a medium-filled Mylar dish will not be irradiated by the alpha particles. The results of the cytogenetic assay of unirradiated target cells that were attached to the top Mylar layer indicate that the number of chromatid-type aberrations was higher when there was a bottom layer of cells in the medium filled chambers than with just medium alone. Furthermore, when the medium was transferred from these cell-irradiated dishes to fresh A(L) cell cultures, chromatid-type aberrations were produced in the unirradiated fresh cells. In contrast, medium irradiated in the absence of cells had no effect on chromatid aberrations. These results suggest that certain unidentified modulating factors secreted from the irradiated cells on the bottom Mylar layer into the medium induce chromatin damage in the unirradiated bystander cells. PMID- 15332999 TI - Risk of thyroid cancer in the Bryansk Oblast of the Russian Federation after the Chernobyl Power Station accident. AB - This population-based case-control study investigated whether exposure to radiation from the Chernobyl Power Station accident is associated with an increased risk of thyroid cancer in children and adolescents aged 0-19 years at the time of the accident who were residing in the more highly contaminated areas of the Bryansk Oblast. Cases were diagnosed with thyroid cancer before October 1, 1997 (n = 26); two controls per case were identified from the Russian State Medical Dosimetrical Registry and were matched by gender, birth year, and raion of residence and type of settlement (urban, town, rural) on April 26, 1986 (n = 52). Individual radiation doses to the thyroid were estimated using a semi empirical model and data were collected in interviews, primarily of the participants' mothers. Based on a loglinear dose-response model treating estimated dose as a continuous variable, the trend of increasing risk with increasing dose was statistically significant (one-sided P = 0.009). These data suggest that exposure to radiation from Chernobyl is associated with an increased risk of thyroid cancer, and that the relationship is dependent on dose. These findings are consistent with descriptive reports from contaminated areas of Ukraine and Belarus, and the quantitative estimate of thyroid cancer risk is generally consistent with estimates from other radiation-exposed populations. PMID- 15333000 TI - Bone marrow origin of cells with capacity for homing and differentiation to esophageal squamous epithelium. AB - Our goal was to determine whether esophageal progenitor cells could be isolated from adult mouse esophagus or bone marrow and shown to home to and proliferate in the irradiated esophagus of recipient mice. Esophageal progenitor cells were isolated from adult male C3H/HeNsd or C57BL/6J green fluorescent protein (GFP(+)) mice by a serial in vitro preplate technique or the technique of side population cell sorting. When injected intravenously (i.v.), these cells homed to the 30-Gy irradiated esophagus of GFP(-) female recipient mice and formed donor-origin esophageal foci. GFP(+) whole murine bone marrow cells injected i.v. also formed donor-origin esophageal squamous cell foci and protected recipient GFP(-) mice from upper-body irradiation in a cell dose-dependent manner. Marrow chimeric GFP( ) mice reconstituted with GFP(+) cells showed migration of GFP(+) marrow cells to the esophagus after 30 Gy irradiation. Purified esophageal progenitor cells isolated from first-generation preplate cell recipients engrafted after i.v. injection to the esophagus of second-generation-irradiated recipient mice. These data establish that esophageal progenitor cells can home to the irradiated esophagus and show limited differentiation capacity to squamous epithelium. PMID- 15333001 TI - Effects of radiation on the environment: a need to question old paradigms and enhance collaboration among radiation biologists and radiation ecologists. AB - A historical perspective is given of the current paradigm that does not explicitly protect nonhuman biota from radiation but instead relies on the concept that if dose limits are set to protect humans, then the environment is automatically protected as well. We summarize recent international questioning of this paradigm and briefly present three frameworks for protecting biota that are being considered by the U.S. Department of Energy, the Canadian Nuclear Safety Commission, and the International Commission on Radiological Protection. We point out a controversial component in each of the three frameworks and suggest topics that need additional research. We emphasize that to properly address radiation protection of the environment, we need to understand how effects are integrated across different levels of biological organization. We caution that the proposed use of molecular end points to estimate ecological risks from radioactive contamination is applicable only if we understand the extent of the impact that molecular damage has on individual organisms and populations of exposed biota. To accomplish the latter, enhanced collaborations are required among the traditionally separate disciplines of radiation biology and radiation ecology. PMID- 15333002 TI - Comparison of microdosimetric simulations using PENELOPE and PITS for a 25 keV electron microbeam in water. AB - The calculations presented compared the performances of two Monte Carlo codes used for the estimation of microdosimetric quantities: Positive Ion Track Structure code (PITS) and a main user code based on the PENetration and Energy Loss of Positrons and Electrons code (PENELOPE-2000). Event-by-event track structure codes like PITS are believed to be superior for microdosimetric applications, and they are written for this purpose. PITS tracks electrons in water down to 10 eV. PENELOPE is one of the few general-purpose codes that can simulate random electron-photon showers in any material for energies from 100 eV to 1 GeV. The model used in the comparison is a water cylinder with an internal scoring geometry made of spheres 1 microm in diameter where the scoring quantities are calculated. The source is a 25 keV electron pencil beam impinging normally on the sphere surface. This work shows only the lineal energy y and spectra graphical presentation as a function of y since for microdosimetry and biology applications, and for discussion of radiation quality in general, these results are more appropriate. The computed PENELOPE results are in agreement with those obtained with the PITS code and published previously in this journal. This paper demonstrates PENELOPE's usefulness at low energies and for small geometries. What is still needed are experimental results to confirm these analyses. PMID- 15333003 TI - Optimization of the auxiliary ligand shell of Cobalt(III)(8-hydroxyquinoline) complexes as model hypoxia-selective radiation-activated prodrugs. AB - A potential approach for activating prodrugs in hypoxic regions of tumors is to use ionizing radiation, rather than bioreductive enzymes, to effect reduction. This study investigates radiolytic release of 8-hydroxyquinoline (8-HQ), as a model for hydroxyaza-chloromethylbenzindoline DNA minor groove alkylators, from Co(III) complexes under hypoxia. 8-HQ release, measured by HPLC, showed higher efficiency (one-electron stoichiometry) when the auxiliary ligand was a tetraazamacrocycle [e.g. 1,4,7,10-tetraazacyclododecane (cyclen)] rather than a triazamacrocycle [1,4,7-triazacyclononane (TACN)]. These complexes differ from the bioreductive cobalt complex SN 24771 in that their reduction provides stable cobalt-containing products rather than free (aquated) Co(2+). Radiolytic release of 8-HQ from Co(cyclen)(8-HQ) and Co(TACN)(CN)(8-HQ) was also demonstrated in deoxygenated human plasma, selectively in the absence of oxygen, again with higher efficiency for the cyclen system. The cobalt complexes were >1000-fold less potent than free 8-HQ as inhibitors of cell proliferation and were metabolically stable in aerobic and hypoxic cell cultures. Investigation of cell uptake of total cobalt, by inductively coupled plasma mass spectrometry, showed that these complexes enter cells but do not accumulate to the high concentrations seen with SN 24771. The results demonstrate the feasibility of masking the cytotoxicity of hydroxyquinoline-based cytotoxins as Co(III) complexes and demonstrate the utility of cyclen-based auxiliary ligands for optimizing radiolytic activation of these novel prodrugs under hypoxia. PMID- 15333004 TI - Dose and dose-rate effects of low-dose ionizing radiation on activation of Trp53 in immortalized murine cells. AB - A derivative of immortalized murine NIH/PG13Luc cells stably transfected with a Trp53-dependent luciferase reporter plasmid was used to study the transcriptional activity of Trp53 in response to radiation. The cell line was sensitive enough to detect the response of Trp53 to 0.2 cGy of (60)Co gamma radiation. To examine the biological effects of low-dose-rate (60)Co gamma radiation (from 0.1-10 cGy/h), we have analyzed the cell cycle, Trp53 transcriptional activity, and gene expression profiles of control and treated cells. Microarray analysis revealed up regulation of six Trp53-mediated genes (Cdkn1a/ p21, Mdm2, Sip27, Ccng1/cyclin G1, Ei24/Pig8 and Dinb/ Polk) after exposure of cells to low-dose-rate radiation for 72 h. Using real-time PCR, a significant elevation in the expression of Ccng1/cyclin G1, Mdm2 and Cdkn1A/p21 was observed with low-dose-rate irradiation at dose rates over 5 cGy/ h. A dose-rate dependence was also observed for these three Trp53-mediated genes. The expression of Ccng1/cyclin G1 at high dose rates of gamma rays was higher than that for low dose rate. However, the expression of Mdm2 for low-dose-rate gamma rays was higher than for the high dose rate. Cells irradiated at low dose rates of 0.1 cGy/h and 1 cGy/h underwent G(1)-phase arrest. Furthermore, G(2)-phase growth arrest was observed in cells irradiated at the low dose rates of 5 cGy/h and 10 cGy/h, which correlated with Trp53-mediated Ccng1/cyclin G1 up-regulation. These results show that cellular response to radiation depended on the dose rate used; i.e., the responses seen at dose rates from 0.1-1 cGy/h were different from those observed at dose rates over 5 cGy/h. PMID- 15333020 TI - The cellular basis of albuminuria. AB - The appearance of albumin in the urine has long been recognized as a cardinal feature of kidney disease and more recently has been shown to also be an independent cardiovascular risk factor associated with insulin resistance. Recent studies on rare human genetic variants, targeted gene disruption in mouse models and cultured glomerular cells in vitro have dramatically improved our understanding of the cellular and molecular basis of albuminuria. This review aims to summarize the current state of knowledge, to illustrate known mechanisms of proteinuria in disease states and to suggest a possible explanation for the link between albuminuria and insulin resistance. PMID- 15333021 TI - Proteolytic cleavage of von Willebrand factor by ADAMTS-13 prevents uninvited clumping of blood platelets. PMID- 15333022 TI - A journey from sickle cell anemia to ADAMTS13. PMID- 15333023 TI - Defective processing of unusually large von Willebrand factor multimers and thrombotic thrombocytopenic purpura. PMID- 15333024 TI - Inherited thrombophilia and life-time risk of venous thromboembolism: is the burden reducible? PMID- 15333025 TI - Familial thrombophilia and lifetime risk of venous thrombosis. AB - BACKGROUND: We started a large multicenter prospective follow-up study to provide reliable risk estimates of venous thrombosis in families with various thrombophilic defects. OBJECTIVES: This paper describes data collected at study entry on venous events experienced before study inclusion, i.e. the baseline data. PATIENTS/METHODS: All individuals (probands, relatives) registered in nine European thrombosis centers with the factor (F)V Leiden mutation, a deficiency of antithrombin, protein C or protein S, or a combination of these defects, were enrolled between March 1994 and September 1997. As control individuals, partners, friends or acquaintances of the thrombophilic participants were included. Incidence and relative risk of objectively confirmed venous thrombotic events (VTEs) prior to entry were calculated for the relatives with thrombophilia and the controls. RESULTS: Of the 846 relatives with thrombophilia (excluding probands), 139 (16%) had experienced a VTE with an incidence of 4.4 per 1000 person years. Of the controls, 15 of the 1212 (1%) controls had experienced a VTE with an incidence of 0.3 per 1000 person years. The risk of venous thrombosis associated with familial thrombophilia was 15.7 (95% CI 9.2-26.8) and remained similar after adjustment for regional and sex-effects (16.4; 95% CI 9.6-28.0). The highest incidence per 1000 person years was found in relatives with combined defects (8.4; 95% CI 5.6-12.2), and the lowest incidence was found in those with the FV Leiden mutation (1.5; 95% CI 0.8-2.6). CONCLUSIONS: Considerable differences in the lifetime risk of VTE were observed among individuals with different thrombophilia defects. PMID- 15333026 TI - The biphasic waveform in plasma: identifying the sepsis-coagulation crossroad? PMID- 15333027 TI - Temporal changes in factors associated with neutrophil elastase and coagulation in intensive care patients with a biphasic waveform and disseminated intravascular coagulation. AB - Summary. The biphasic waveform is an early marker of disseminated intravascular coagulation (DIC). Neutrophil elastase (NE) cleaves coagulation factors; thus, elevated elastase levels or its dysregulation by alpha-1-protease inhibitor (Alpha1PI) may be linked to DIC. Time courses over a period were determined for factors associated with NE and coagulation in 14 Intensive Care Unit patients with a biphasic waveform who developed DIC. The data were analyzed using a random coefficient linear regression model to predict the variables' mean values on day 0 and their mean rates of change over the period in which the biphasic waveform appeared. The biphasic waveform was normal on day 0, maximized on day 1, and approached normal again by day 4. Alpha1PI/NE complex levels were 2.5-fold greater than normal for the entire period. The A1PI activity, antigen, and specific activity levels were normal on day 0 and increased thereafter by 21.0, 10.5, and 8.9% of normal per day, respectively. Factor II, V, VII, IX, and X activity levels were, respectively, 57, 46, 46, 77, and 46% of normal on day 0, whereas factor VIII and fibrinogen levels were normal. All coagulation factor levels trended upward with time but not significantly. The prothrombin time, but not the activated partial thromboplastin time, was prolonged, and the platelet counts and hematocrits were below normal on day 0 and remained so thereafter. We conclude that events associated with neutrophil activation, elastase release, and perturbations of coagulation precede both the appearance of the biphasic waveform and the diagnosis of DIC in these patients. PMID- 15333028 TI - Packaging is important: accelerated thrombolysis with encapsulated plasminogen activators. PMID- 15333029 TI - Distributed intraclot thrombolysis: mechanism of accelerated thrombolysis with encapsulated plasminogen activators. AB - BACKGROUND: The delivery of encapsulated plasminogen activators has demonstrated enhanced thrombolysis in vivo in several models. The mechanism of such improvement has not previously been established. OBJECTIVES: We explored in vitro the mechanism by which microencapsulation of streptokinase in polymeric microparticles accelerates clot digestion and reduces reperfusion times by as much as an order of magnitude in vivo. METHODS: The efficacy of microencapsulated streptokinase (MESK) was directly compared with identical dosages of unencapsulated streptokinase (FREE SK) at three initial pressure drops using clots formed of plasma or whole blood in 0.2-cm inner diameter glass capillary tubes. RESULTS: MESK demonstrated accelerated flow restoration compared with FREE SK for each condition in plasma (23.8 +/- 4.5% faster) and whole blood clots (17.2 +/- 9.2% faster). Images collected by light microscopy show sites of thrombolysis internal to the clot only with MESK while the spatial distribution of fluorescently labeled streptokinase by confocal microscopy confirms greater penetration of the encapsulated agent compared with unencapsulated streptokinase. Digestion thus proceeds in three dimensions rather than restricted to a two dimensional lysis front. CONCLUSIONS: The improved clot penetration with MESK establishes enhanced transport with encapsulation and the concept of distributed intraclot thrombolysis as a basis for the accelerated dissolution observed with encapsulated plasminogen activators in vivo. PMID- 15333030 TI - New treatment option for the antiphospholipid antibody syndrome? More pleiotropic effects of the statin drugs. PMID- 15333031 TI - Fluvastatin inhibits up-regulation of tissue factor expression by antiphospholipid antibodies on endothelial cells. AB - BACKGROUND: Mechanisms of thrombosis induced by antiphospholipid (aPL) antibodies include up-regulation of tissue factor (TF) expression on endothelial cells (ECs). Statins have been shown to reduce levels of TF induced by tumor necrosis factor (TNF-alpha) and lipopolysaccharide (LPS) on ECs. In a recent study, fluvastatin inhibited thrombogenic and proinflammatory properties of aPL antibodies in in vivo models. The aim of this study was to determine whether fluvastatin has an effect on aPL-induced expression of TF on ECs. METHODS: IgGs were purified from four patients with APS (IgG-APS) and from control sera (IgG NHS). Cultured human umbilical vein endothelial cells (HUVEC) were treated with IgG-APS or IgG-NHS or with medium alone or with phorbol myristate acetate (PMA), as a positive control. In some experiments, cells were pretreated with fluvastatin (2.5, 5 or 10 micro m) with and without mevalonate (100 micro m). TF expression on HUVECs was measured by ELISA. RESULTS: PMA and the four IgG-APS preparations increased the expression of TF on EC significantly (4.9-, 2.4-, 4.2 , 3.5- and 3.1-fold, respectively), in a dose-dependent fashion. Fluvastatin (10 micro m) inhibited the effects of PMA and the four IgG-APS on TF expression by 70, 47, 65, 22 and 68%, respectively, and this effect was dose-dependent. Mevalonate (100 micro m) completely abrogated the inhibitory effects of fluvastatin on TF expression induced by aPL. CONCLUSION: Because of the suggested pathogenic role of aPL on induction of TF on ECs, our data provide a rationale for using statins as a therapeutic tool in treatment of thrombosis in APS. PMID- 15333032 TI - Familial multiple coagulation factor deficiencies: new biologic insight from rare genetic bleeding disorders. AB - Combined deficiency of factor (F)V and FVIII (F5F8D) and combined deficiency of vitamin K-dependent clotting factors (VKCFD) comprise the vast majority of reported cases of familial multiple coagulation factor deficiencies. Recently, significant progress has been made in understanding the molecular mechanisms underlying these disorders. F5F8D is caused by mutations in two different genes (LMAN1 and MCFD2) that encode components of a stable protein complex. This complex is localized to the secretory pathway of the cell and likely functions in transporting newly synthesized FV and FVIII, and perhaps other proteins, from the ER to the Golgi. VKCFD is either caused by mutations in the gamma-carboxylase gene or in a recently identified gene encoding the vitamin K epoxide reductase. These two proteins are essential components of the vitamin K dependent carboxylation reaction. Deficiency in either protein leads to under-carboxylation and reduced activities of all the vitamin K-dependent coagulation factors, as well as several other proteins. The multiple coagulation factor deficiencies provide a notable example of important basic biological insight gained through the study of rare human diseases. PMID- 15333033 TI - Dose escalating safety study of a new oral direct thrombin inhibitor, dabigatran etexilate, in patients undergoing total hip replacement: BISTRO I. AB - BACKGROUND: Dabigatran etexilate (BIBR 1048) is an oral direct thrombin inhibitor undergoing evaluation for the prevention of venous thromboembolism (VTE) following total hip replacement. Following oral administration, dabigatran etexilate is rapidly converted to its active form dabigatran (BIBR 953 ZW). OBJECTIVES: To determine the safe therapeutic range of dabigatran etexilate following total hip replacement. METHODS: In a multicenter, open-label, dose escalating study, 314 patients received oral doses of dabigatran etexilate (12.5, 25, 50, 100, 150, 200 and 300 mg twice daily or 150 and 300 mg once daily) administered 4-8 h after surgery, for 6-10 days. Dose escalation was based on clinical and pharmacokinetic data. The primary safety outcome was major bleeding. The primary efficacy outcome included venographic deep vein thrombosis (DVT), symptomatic DVT and pulmonary embolism, during the treatment period. RESULTS: No major bleeding event was observed in any group, but two patients at the highest dose (300 mg twice daily) suffered bleeding from multiple sites associated with reduced renal clearance and prolonged pharmacodynamic (PD) parameters. A dose response was demonstrated for minor bleeding events. Of the 289 treated patients, 225 patients had evaluable venograms. The overall incidence of DVT was 12.4% (28/225 patients). There was no consistent relationship between the dose and incidence of DVT, the highest incidence in any group being 20.8% (5/24 patients). The lowest dose (12.5 mg twice daily) showed a high rate of proximal DVT [12.5% (3/24)] and no increase in PD parameters. Peak and trough plasma concentrations, area under the dabigatran plasma concentration-time curve and PD parameters also increased in proportion with the dose. Higher dabigatran plasma concentrations were associated with lower DVT rates. Approximately 20% of the patients had low plasma concentrations after the first dose suggesting further optimization of the preliminary tablet formulation is required. CONCLUSIONS: Dabigatran etexilate demonstrates an acceptable safety profile, with a therapeutic window above 12.5 mg and below 300 mg twice daily. The low number of VTE events within each treatment group indicates a satisfactory antithrombotic potential, although the study was not powered for an efficacy analysis. Additional studies are ongoing to optimize oral absorption and the efficacy/safety balance. PMID- 15333034 TI - Correlation between thrombus regression and recurrent venous thromboembolism. Examining venographic and clinical effects of low-molecular-weight heparins: a meta-analysis. AB - We analyzed the correlation between thrombus regression on control venography performed after discontinuation of heparin therapy and recurrent venous thromboembolism (VTE) detected during clinical follow-up in randomized trials comparing low-molecular-weight heparin (LMWH) and unfractionated heparin (UFH) in patients with deep vein thrombosis (DVT). Data were abstracted from MEDLINE, conference abstracts and reference lists of previous reviews. Randomized, controlled trials comparing LMWH and UFH for the treatment of DVT using a combined venographic and clinical assessment and with at least 2 months of follow up were selected. The proportions of patients with thrombus regression on control venography performed soon after discontinuation of heparin therapy and recurrent VTE at 2-6 months were independently collected by two researchers. Thirteen studies met the inclusion criteria. There was a strong inverse correlation between thrombus regression and recurrent VTE (r =- 0.70; P =0.008). The venographic effect varied between the different LMWHs (P = 0.013). A very strong correlation was found when the results were pooled by the type of LMWH used (r = 0.84; P=0.037). No influence of the dose interval used on the venographic effect (P=0.156) or on recurrent VTE (P=0.218) was shown. The lack of thrombus regression in venography, performed soon after heparin discontinuation, was correlated with clinical recurrence. Non-invasive imaging techniques should be relevant to identify non-responders and to assess the optimal duration of initial heparin treatment in daily clinical practice. PMID- 15333035 TI - Preeclampsia and its interaction with common variants in thrombophilia genes. AB - Recently, it has been proposed that abnormalities in coagulation and fibrinolysis contribute to the development of preeclampsia by increasing the thrombotic tendency. This hypothesis was tested in women who have had preeclampsia (cases) compared with matched controls. Polymorphisms in the thrombophilia genes [plasminogen activator inhibitor type 1 [PAI-1 -675(4G/5G)], thrombin activatable fibrinolysis inhibitor (TAFI -438G/A and 1040C/T), methylenetetrahydrofolate reductase (MTHFR 677C/T), factor V (FV Leiden R/Q506), prothrombin (FII 20210G/A) and factor XIIIA (FXIIIA V/L34)] were determined in 157 women with preeclampsia and 157 women with uncomplicated pregnancy. The associated risk of preeclampsia was analyzed using logistic regression methods. The frequency distributions of the genotypes of these six polymorphisms in thrombophilia genes were similar in the case and control groups. We found no differences in the prevalence of genetic risk factors of thrombosis in women with preeclampsia compared with controls, which makes it unlikely that these polymorphisms are risk factors for preeclampsia. PMID- 15333036 TI - Impact of progestagens on activated protein C (APC) resistance among users of oral contraceptives. AB - Oral contraceptive (OC) use is associated with an increased risk of venous thromboembolism. Previous data reported higher thrombotic risk in women using third-generation combined OC than in those using second generation OC. The difference could be explained by differential effects of progestagens on plasma sensitivity to activated protein C (APC). The main purpose of this cross sectional study was to assess the influence of a progestagen-only OC (chlormadinone acetate) as well as the effect of several combined OC with different progestagen components on APC resistance. The effect of APC on endogenous thrombin potential (ETP) was investigated in the plasma of healthy women using either combined OC (n=82) or progestagen-only OC (n=28), and in non users (n=64). Carriers of factor V Leiden were excluded. Compared with non-users, there was no significant change in APC resistance in women using progestagen-only OC. Women who used combined OC were less sensitive to APC than non-users (P < 0.001) and the difference was significantly more pronounced in women using third generation OC (n=41) than in those who used second-generation OC containing levonorgestrel (n=22) (P < 0.05). Compared with OC containing levonorgestrel, use of norethisterone-containing OC (n = 9) was associated with an increased resistance to APC (P < 0.05). Women who used cyproterone-containing OC (n = 10) were less sensitive to APC than those using third-generation OC (P < 0.05) or second-generation OC containing levonorgestrel (P < 0.05). Protein S, factor II and FVIII levels explained in part the OC-related changes in APC sensitivity variations. ETP-based APC resistance may contribute to explain why different brands of OC can be associated with different levels of thrombogenicity. PMID- 15333038 TI - Properties of recombinant human thromboplastin that determine the International Sensitivity Index (ISI). AB - Prothrombin Time (PT) clotting tests are widely used to monitor oral anticoagulation therapy and to screen for clotting factor deficiencies. The active ingredient in PT reagents (thromboplastins) is tissue factor, the integral membrane protein that triggers the clotting cascade through the extrinsic pathway. Several years ago, a system for calibrating and using thromboplastin reagents, known as the International Sensitivity Index (ISI) and the International Normalized Ratio (INR), was developed to standardize monitoring of oral anticoagulant therapy. The ISI/INR method, while revolutionizing the monitoring of coumarin therapy, has been criticized for a number of perceived shortcomings. We have undertaken a series of studies aimed at achieving a detailed understanding of which parameters influence the ISI values of thromboplastin reagents, with an ultimate goal of creating 'designer thromboplastins' whose sensitivities to the various clotting factors can be individually tailored. In this study, we demonstrate that ISI values of thromboplastin reagents based on relipidated, recombinant human tissue factor can be controlled by a combination of changes in the phospholipid content (in particular, the levels of phosphatidylserine and phosphatidylethanolamine) and ionic strength. The sensitivity of a given thromboplastin reagent can be increased (i.e. its ISI value decreased) by decreasing the content of phosphatidylserine and/or increasing the ionic strength. The molar ratio of phospholipid to tissue factor, on the other hand, had essentially no impact on ISI value. PMID- 15333037 TI - Measurement of von Willebrand factor cleaving protease (ADAMTS-13): results of an international collaborative study involving 11 methods testing the same set of coded plasmas. AB - BACKGROUND: ADAMTS-13 is a von Willebrand factor (VFW)-cleaving protease. Its congenital or acquired deficiency is associated with thrombotic thrombocytopenic purpura (TTP) and more rarely with the hemolytic uremic syndrome. We report on a survey evaluating 11 methods for ADAMTS-13 measurement performed in different labs. DESIGN: Two plasmas, one normal and one from a patient with familial TTP, were mixed at the co-ordinating center to prepare 6 plasmas with 0%, 10%, 20%, 40%, 80% and 100% ADAMTS-13 levels. Each plasma was aliquoted and assembled into sets of 60 (coded from 1 to 60), each containing 10 copies of the original 6 plasmas. Plasmas were frozen and shipped in dry ice to 10 labs with a common frozen reference plasma. Laboratories were asked to measure ADAMTS-13 with their methods. Results were sent to the coordinating center for statistical analysis. RESULTS: Of the 10 methods performed under static conditions 9 were quantitative and one was semiquantitative. One method performed under flow conditions evaluated the extent of cleavage of endothelial cell-derived ultralarge VWF string-like structures and expressed results as deficient, normal, or borderline. Linearity (expected-vs-observed levels), assessed as the squared correlation coefficient, ranged from 0.98 to 0.39. Reproducibility, expressed as the coefficient of variation for repeated measurements, ranged from < 10% to 83%. The majority of methods were able to discriminate between different ADAMTS-13 levels. The majority were able to detect the plasma with 0% level and some of them to discriminate between 0% and 10%. Overall the best performance was observed for three methods measuring cleaved VWF by ristocetin cofactor, collagen binding, and immunoblotting of degraded multimers of VWF substrate, respectively. The poor interlaboratory agreement of results was hardly affected by the use of the common standard. The method performed under flow conditions identified the plasmas with 0%, 10%, 20% and 40% activity as deficient in 7, 5, 1 and 3 of the 10 replicate measurements. The plasmas with 80% and 100% were identified as normal in all of the 10 replicate measurements. CONCLUSIONS: The survey shows varied performance, but supports an optimistic view about the reliability of current methods for ADAMTS-13. PMID- 15333039 TI - Comparative effects of microplasmin and tissue-type plasminogen activator (tPA) on cerebral hemorrhage in a middle cerebral artery occlusion model in mice. AB - BACKGROUND: Thrombolytic therapy of ischemic stroke with tissue-type plasminogen activator (tPA) improves clinical outcome which may, however, be partially offset by significant intracerebral bleeding (ICB). OBJECTS: The comparative effects of microplasmin (microPli) and tPA on ICB were evaluated in a thrombotic middle cerebral artery (MCA) occlusion model in mice. METHODS: A dose of microPli (5 mg kg(-1)) or tPA (4 mg kg(-1)) which are comparably effective for reduction of brain damage, or a double dose (10 or 8 mg kg(-1), respectively) or the microPli excipient as a control were intravenously administered as a bolus at 30 min or 4 h after MCA occlusion. ICB was measured at 24 h by hemoglobin assay of exsanguinated brain extracts. Bleeding time was measured by tail cutting. RESULTS: In controls given solvent at 4 h, ICB was on average 8.8 micro L, which was significantly increased with 10 mg kg(-1) microPli and with 4 and 8 mg kg(-1) tPA to 12-13 micro L (P < 0.05 each vs. controls, n = 7-9), whereas 5 mg kg(-1) microPli did not affect bleeding (8.5 micro L P = NS vs. controls, n = 7). When given at 30 min, neither microPli nor tPA altered ICB (6.3-6.8 micro L, mean; n = 7-9). tPA but not microPli increased bleeding time; from 2.4 min in controls to 5.9 min (median, P < 0.05 vs. controls) and 8.7 min (P < 0.01 vs. controls) with 4 and 8 mg kg(-1), respectively, and to 2.3 and 2.9 min with 5 and 10 mg kg(-1) microPli, respectively (n = 10). CONCLUSIONS: microPli at a dose comparably effective as tPA for brain damage reduction induced significantly less ICB, and less bleeding time prolongation in mice with thrombotic MCA occlusion. PMID- 15333040 TI - A monoclonal antibody directed against human von Willebrand factor induces type 2B-like alterations. AB - We have previously described a monoclonal antibody (mAb), 1C1E7, against von Willebrand factor (VWF), that increases ristocetin-induced platelet aggregation (RIPA) and induces a preferential binding of the high-molecular-weight multimers of VWF to platelet GPIb. Further investigations using a rotational viscometer at a shear rate of 4000 s(-1) could now demonstrate that shear-induced platelet aggregation (SIPA) is significantly increased with 1C1E7 and that this could be completely inhibited by the anti-GPIb mAb 6D1. In contrast, platelet adhesion to a collagen surface at a shear rate of 2600 s(-1), using a rectangular perfusion chamber, was significantly inhibited in the presence of 1C1E7. When citrated whole blood was incubated with 1C1E7, a spontaneous binding of VWF to the platelet GPIb could be demonstrated by flow cytometric analysis. Parallel to this, a decrease of the highest molecular weight multimers of VWF in the plasma was found. Platelets with bound VWF on their surface were able to form macroaggregates but were no longer able to adhere. These phenomena are very similar to the alterations described in von Willebrand's disease type 2B. The epitope of this mAb could be localized to the N-terminal part of the subunit; therefore a distant conformational change in the A1 domain of VWF is suggested. PMID- 15333041 TI - Fibrinogen binding potentiates FGF-2 but not VEGF induced expression of u-PA, u PAR, and PAI-1 in endothelial cells. AB - Endothelial cell responses at sites of injury occur in a fibrin matrix and are regulated by growth factors including those of the FGF and VEGF families. The pericellular proteolytic balance is important in these responses, and FGF-2 and VEGF up-regulate endothelial cell u-PA, u-PAR and PAI-1. Because both VEGF and FGF-2 bind to fibrinogen, we have examined the capacity of fibrinogen to modulate the up-regulation of these proteins by FGF-2 and VEGF. Confluent cultures of endothelial cells were exposed to FGF-2, VEGF, and fibrinogen or to combinations of growth factors with fibrinogen. Changes in mRNA levels of u-PA, u-PAR and PAI 1 were measured by Northern blot. FGF-2 increased u-PA, u-PAR, and PAI-1 mRNA, but there was a significantly greater induction when fibrinogen was added to FGF 2 at all concentrations. The potentiation by fibrinogen was particularly evident at an FGF-2 concentration of 0.1 ng mL(-1), which resulted in non-significant change in transcript levels by itself, but significantly increased up to 2.6-fold with fibrinogen. VEGF also increased endothelial cell expression of u-PA, u-PAR and PAI-1, but this effect was not potentiated by fibrinogen. Addition of LM609, a monoclonal antibody to alphaVbeta3, significantly inhibited induction of u-PA mRNA and activity by fibrinogen-bound FGF-2 compared to FGF-2. A monoclonal antibody to FGFR1 also inhibited u-PA mRNA expression induced by fibrinogen-bound FGF-2. We conclude that fibrinogen increases the capacity of FGF-2, but not of VEGF, to up-regulate u-PA, u-PAR, and PAI-1 in endothelial cells and that fibrinogen-bound FGF-2 requires alphaVbeta3 binding to up-regulate endothelial cell u-PA. PMID- 15333042 TI - Atrial fibrillation and thrombosis: immunohistochemical differences between in situ and embolized thrombi. AB - BACKGROUND/OBJECTIVE: Thromboembolism secondary to atrial fibrillation accounts for approximately one-fourth of all strokes. Although considerable resources have been targeted to pharmacologic prophylaxis, neither the cellular nor the biochemical composition of atrial thrombi is known. Quantitative immunohistochemistry was undertaken to define the composition of atrial thrombi and to explore morphological differences between atrial appendage thrombi and those that embolize. PATIENTS/METHODS: Serial sections of thrombi obtained during valve replacement surgery or embolectomy from 22 patients with atrial fibrillation were stained with antibodies against fibrin, integrin beta3, or tissue factor and analyzed with NIH-image. RESULTS: Thrombi showed distinct regions staining for either fibrin or platelets and on average, the fibrin-rich regions predominated (P < 0.0001). The platelet content of embolized thrombi was nearly twice that of atrial thrombi (P = 0.02). Non-staining amorphous material comprised nearly half of atrial thrombi in situ, but was rare in embolized thrombi (P < 0.001). Tissue factor colocalized to areas rich in platelets and granulocytes. CONCLUSIONS: The abundance of fibrin relative to platelets underscores the enhanced efficacy of warfarin prophylaxis in clinical trials. The finding of tissue factor localized to platelet-leukocyte clusters suggests its blood-borne origin. Compositional differences between in situ and embolized thrombi suggest directions for investigating propensity for embolization. PMID- 15333043 TI - Extracellular matrix molecules regulate endothelial cell migration stimulated by lysophosphatidic acid. AB - BACKGROUND: Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are lipids that bind G-protein coupled receptors and differentially promote transmigration of endothelial cells. OBJECTIVE: To determine if endothelial cell transmigration stimulated by LPA, not S1P, is dependent on the extracellular matrix. METHODS: Bovine pulmonary artery (BPAE) endothelial cell transmigration and locomotion were measured using a modified-Boyden chamber and video microscopy, respectively. Results were related to strength of adhesion and characteristics of cell adhesive contacts. RESULTS AND CONCLUSIONS: BPAEs responded to LPA by transmigration through gelatin- or collagen-coated filters, but not through fibronectin-, vitronectin-, or fibrinogen-coated filters. Fewer cells adhered to collagen or gelatin than to fibronectin in a static cell adhesion assay or after application of a g-force to detach cells. Video microscopy revealed that S1P stimulates large lamellipodia on two-dimensional fibronectin substrate. LPA stimulated lamellipodia on fibronectin, but the trailing edge remained attached, resulting in sting ray-shaped cells in video microscopy. LPA-treated cells on gelatin released the trailing edge. To understand how the extracellular matrix may regulate endothelial cell shape during movement, we surveyed changes in focal adhesion proteins. More Hic-5, a paxillin homolog, was detected in the detergent insoluble fraction of BPAEs attached to gelatin than fibronectin. No such difference was found in paxillin. In BPAEs, Hic-5 was localized to smaller punctate structures on fibronectin and longer, thinner focal adhesions on gelatin. These results indicated that localization of Hic-5 and strength of adhesion correlate with endothelial cell transmigration stimulated by LPA, but not with transmigration stimulated by S1P. PMID- 15333044 TI - Cytokine gene expression and production by human LPS-stimulated mononuclear cells are inhibited by sulfated heparin-like semi-synthetic derivatives. AB - BACKGROUND: The K5 polysaccharide obtained from Escherichia coli strain 010:K5:H4 is a polymer of the disaccharidic unit formed by D-glucuronic acid and N acetylglucosamine. This structure is akin to N-acetylheparosan, the precursory polymer of heparin and of heparan sulfate. This structural affinity with N acetylated heparin and with de-sulfated heparin makes the K5 polysaccharide extremely useful for the preparation of sulfated heparin-like semi-synthetic derivatives. It has been demonstrated that heparins are able to inhibit tissue factor and cytokine production and expression by human monocytes. OBJECTIVE: The aim of this study was to evaluate the effects of four different heparin-like semi synthetic derivatives on inflammatory cytokine production and expression by human mononuclear cells. RESULTS: The simultaneous addition of lipopolysaccharide (LPS; 0.2 and 10 micro g mL(-1)) and the K5 polysaccharide did not inhibit interleukin (IL)-1beta, IL-6 or tumor necrosis factor (TNF)-alpha production by stimulated mononuclear cells. IL-1beta, IL-6 and TNF-alpha concentrations in supernatants of LPS-stimulated mononuclear cells were not influenced by the addition of N,O sulfated K5 polysaccharide (K5-N, OS) and epimerized N-sulfated K5 polysaccharide (K5 NS epi) at 5 and 10 microg mL(-1), whereas the addition of epimerized N,O sulfated K5 polysaccharide (K5-N, OS epi) (5 and 10 microg mL(-1)) and O-sulfated K5 polysaccharide (K5-OS) (5 and 10 microg mL(-1)) to LPS-stimulated cells caused a significant dose-dependent inhibition of IL-1beta, IL-6 and TNF-alpha. All sulfated heparin-like semi-synthetic derivatives did not influence the IL-10 production by LPS-stimulated mononuclear cells. In LPS-stimulated cells (0.2 and 10 microg mL(-1)), K5-OS or K5-N, OS epi at 5 and 10 microg mL(-1) markedly decreased TNF-alpha mRNA expression. CONCLUSIONS: These results indicate that the sulfated heparin-like semi-synthetic derivatives K5-OS and K5-N, OS epi are able to inhibit both expression and production of inflammatory cytokines, whereas they do not influence the anti-inflammatory cytokine IL-10, suggesting a potential role for these products as modulators of inflammatory reactions. PMID- 15333045 TI - A 13 base pair deletion in the GPIbbeta gene in a second unrelated Bernard Soulier family due to slipped mispairing between direct repeats. PMID- 15333046 TI - Treatment of heparin induced thrombocytopenia and thrombosis during the first trimester of pregnancy. PMID- 15333047 TI - Estimated risk of deep venous thrombosis among Chinese patients admitted to general medical wards. PMID- 15333048 TI - The 4G-allele of the PAI-1 gene is not consistently associated with a higher prevalence of coronary stenosis. PMID- 15333049 TI - d-Dimer testing to predict recurrence risk in venous thromboembolism: looking for a useful threshold: a rebuttal. PMID- 15333053 TI - Is physician self-disclosure related to patient evaluation of office visits? AB - CONTEXT: Physician self-disclosure has been viewed either positively or negatively, but little is known about how patients respond to physician self disclosure. OBJECTIVE: To explore the possible relationship of physician self disclosure to patient satisfaction. DESIGN: Routine office visits were audiotaped and coded for physician self-disclosure using the Roter Interaction Analysis System (RIAS). Physician self-disclosure was defined as a statement describing the physician's personal experience that has medical and/or emotional relevance for the patient. We stratified our analysis by physician specialty and compared patient satisfaction following visits in which physician self-disclosure did or did not occur. PARTICIPANTS: Patients (N= 1,265) who visited 59 primary care physicians and 65 surgeons. MAIN OUTCOME MEASURE: Patient satisfaction following the visit. RESULTS: Physician self-disclosure occurred in 17% (102/589) of primary care visits and 14% (93/676) of surgical visits. Following visits in which a primary care physician self-disclosed, fewer patients reported feelings of warmth/friendliness (37% vs 52%; P =.008) and reassurance/comfort (42% vs 55%; P =.027), and fewer reported being very satisfied with the visit (74% vs 83%; P =.031). Following visits in which a surgeon self-disclosed, more patients reported feelings of warmth/friendliness (60% vs 45%; P =.009) and reassurance/comfort (59% vs 47%; P=.044), and more reported being very satisfied with the visit (88% vs 75%; P =.007). After adjustment for patient characteristics, length of the visit, and other physician communication behaviors, primary care patients remained less satisfied (adjusted odds ratio [AOR], 0.45; 95% confidence interval [CI], 0.24 to 0.81) and surgical patients more satisfied (AOR, 2.22; 95% CI, 1.12 to 4.50) after visits in which the physician self-disclosed. CONCLUSIONS: Physician self-disclosure is significantly associated with higher patient satisfaction ratings for surgical visits and lower patient satisfaction ratings for primary care visits. Further study is needed to explore these intriguing findings and to define the circumstances under which physician self-disclosure is either well or poorly received. PMID- 15333054 TI - What do physicians tell patients about themselves? A qualitative analysis of physician self-disclosure. AB - OBJECTIVE: Physician self-disclosure (PSD) has been alternatively described as a boundary violation or a means to foster trust and rapport with patients. We analyzed a series of physician self-disclosure statements to inform the current controversy. DESIGN: Qualitative analysis of all PSD statements identified using the Roter Interaction Analysis System (RIAS) during 1,265 audiotaped office visits. SETTING AND PARTICIPANTS: One hundred twenty-four physicians and 1,265 of their patients. MAIN RESULTS: Some form of PSD occurred in 195/1,265 (15.4%) of routine office visits. In some visits, disclosure occurred more than once; thus, there were 242 PSD statements available for analysis. PSD statements fell into the following categories: reassurance (n = 71), counseling (n = 60), rapport building (n = 55), casual (n = 31), intimate (n = 14), and extended narratives (n = 11). Reassurance disclosures indicated the physician had the same experience as the patient ("I've used quite a bit of that medicine myself"). Counseling disclosures seemed intended to guide action ("I just got my flu shot"). Rapport building disclosures were either humorous anecdotes or statements of empathy ("I know I'd be nervous, too"). Casual disclosures were short statements that had little obvious connection to the patient's condition ("I wish I could sleep sitting up"). Intimate disclosures refer to private revelations ("I cried a lot with my divorce, too") and extended narratives were extremely long and had no relation to the patient's condition. CONCLUSIONS: Physician self-disclosure encompasses complex and varied communication behaviors. Self-disclosing statements that are self-preoccupied or intimate are rare. When debating whether physicians ought to reveal their personal experiences to patients, it is important for researchers to be more specific about the types of statements physicians should or should not make. PMID- 15333055 TI - Prevalence of nutrition and exercise counseling for patients with hypertension. United States, 1999 to 2000. AB - OBJECTIVE: To evaluate the prevalence of nutrition and exercise counseling for patients with hypertension. DESIGN: Cross-sectional survey data from the National Ambulatory Medical Care Survey (NAMCS) and the National Hospital Ambulatory Medical Care Survey (NHAMCS) for 1999 and 2000. SETTING: Office-based physician practices and hospital outpatient departments. PATIENTS/PARTICIPANTS: Patients age 18 or older with a diagnosis of hypertension. MEASUREMENTS AND MAIN RESULTS: In 1999 and 2000, over 137 million patient encounters had a diagnosis of hypertension. Nutrition and exercise counseling were provided at 35% and 26% of visits, respectively. Patients older than 74 received the least nutrition (28%) and exercise (18%) counseling. Asians and Hispanics were more likely to undergo any lifestyle counseling, while non-Hispanic whites received the least exercise and nutrition counseling. Patients with 2 cardiovascular comorbidities were counseled for diet (53%) and exercise (32%) more than those with 1 (44% and 31%) or none (30% and 23%; P.05) by race/ethnicity in doctor-level reasons for nonreceipt of recommended invasive cardiac procedures. CONCLUSIONS: Patient decisions to decline recommended invasive cardiac procedures were infrequent and may explain only a small fraction of racial disparities in the use of invasive cardiac procedures. PMID- 15333062 TI - Impact of English language proficiency on receipt of pap smears among Hispanics. AB - Our aim was to assess the impact of English language proficiency on Pap smear use among Hispanics. We performed a cross-sectional study using 2000 National Health Interview Survey data and included 2,331 Hispanic women, age >/=18 without a hysterectomy. After adjusting for sociodemographic and access factors, highly proficient English speakers were more likely to report a Pap smear in the past 3 years as compared to low proficient (adjusted prevalence ratio, 1.16; 95% confidence interval, 1.08 to 1.22). Also associated with Pap smear use were income, usual source of care, and health insurance. Our finding suggests that low English language proficiency is a barrier to receiving recent Pap smears among Hispanics. PMID- 15333063 TI - How reliable are assessments of clinical teaching? A review of the published instruments. AB - BACKGROUND: Learner feedback is the primary method for evaluating clinical faculty, despite few existing standards for measuring learner assessments. OBJECTIVE: To review the published literature on instruments for evaluating clinical teachers and to summarize themes that will aid in developing universally appealing tools. DESIGN: Searching 5 electronic databases revealed over 330 articles. Excluded were reviews, editorials, and qualitative studies. Twenty-one articles describing instruments designed for evaluating clinical faculty by learners were found. Three investigators studied these papers and tabulated characteristics of the learning environments and validation methods. Salient themes among the evaluation studies were determined. MAIN RESULTS: Many studies combined evaluations from both outpatient and inpatient settings and some authors combined evaluations from different learner levels. Wide ranges in numbers of teachers, evaluators, evaluations, and scale items were observed. The most frequently encountered statistical methods were factor analysis and determining internal consistency reliability with Cronbach's alpha. Less common methods were the use of test-retest reliability, interrater reliability, and convergent validity between validated instruments. Fourteen domains of teaching were identified and the most frequently studied domains were interpersonal and clinical-teaching skills. CONCLUSIONS: Characteristics of teacher evaluations vary between educational settings and between different learner levels, indicating that future studies should utilize more narrowly defined study populations. A variety of validation methods including temporal stability, interrater reliability, and convergent validity should be considered. Finally, existing data support the validation of instruments comprised solely of interpersonal and clinical-teaching domains. PMID- 15333064 TI - The emerging role of online communication between patients and their providers. AB - Despite the explosion of online communication in the community, its use between patients and their health care providers remains low. However, rapidly growing patient and provider interest in using online communication has motivated organizations to consider options for deploying these new tools in clinical practice. In this paper, we describe the barriers and challenges health care providers and their organizations must address in developing and deploying these new tools. We formulate lessons from early experiences with e-mail and web-based communication in clinical settings. Finally, we provide a roadmap for developing and deploying these new tools in clinical practice. Health care providers and their organizations will need to consider issues related to technology, data management, operations, communication management, and financial support in order to successfully deploy online services and communication for patients in clinical settings. PMID- 15333065 TI - Is self-disclosure a boundary violation? PMID- 15333066 TI - Health disparities. Toward a better understanding of primary care patient physician relationships. PMID- 15333068 TI - Survey of internal medicine departments on faculty development activities. PMID- 15333070 TI - Australian Gastroenterology Week 2004. Abstracts. PMID- 15333074 TI - Conduction characteristics at the crista terminalis during onset of pulmonary vein atrial fibrillation. AB - INTRODUCTION: Focal atrial fibrillation (AF) may initiate with an irregular rapid burst of atrial ectopic (AE) activity from a pulmonary vein (PV) focus, but how AF is maintained it is not known. The crista terminalis (CT) is an important line of block in atrial flutter (AFL), but its role in AF has not been determined. The aim of this study was to examine the conduction properties of the CT during onset of AF. METHODS AND RESULTS: In 10 patients (mean age 38 +/- 8 years), we analyzed conduction across the CT during onset of focal AF from an arrhythmogenic PV and during pacing from the same PV at cycle lengths of 700 and 300 ms. A 20-pole catheter was positioned on the CT using intracardiac echocardiography. In 10 control patients with no history of AF, we analyzed conduction across the CT during pacing from the distal coronary sinus at 700 and 300 ms. In all 10 AF patients, AF was initiated with 1 to 9 AE beats (median 5) from a PV. During sinus rhythm, there were no split components (SC) recorded on the CT. During PV AE activity, discrete SC were recorded on the CT in all patients over 6.3 +/- 0.9 bipoles (3.7 +/- 0.3 cm). Maximal splitting of SC was 66 +/- 31 ms (37-139). There was an inverse relationship between AE coupling intervals and the degree of splitting between SC in all patients. Degeneration to AF was preceded by progressive decrement across the CT. SC were recorded during PV pacing at 700 and 300 ms (maximal distance between SC of 24 +/- 3 ms and 43 +/- 5 ms, respectively, P < 0.001). Maximum SC at CT in controls was 13 +/- 8 ms at 700 ms (P = 0.06 vs AF patients) and 16 +/- 9 ms at 300 ms (P < 0.01 vs AF patients). CONCLUSION: (1) These observations provide evidence of anisotropic, decremental conduction across the CT during onset of focal AF and during pacing from the same PV. A line of functional conduction block develops along this anatomic structure (CT). Whether this line of block acts as an initiator of AF or simply contributes passively to nonuniform fibrillatory conduction is unknown. (2) In some patients with focal AF, development of conduction block along the CT may provide a substrate for typical AFL. PMID- 15333075 TI - Biventricular implantable cardioverter defibrillators improve survival compared with biventricular pacing alone in patients with severe left ventricular dysfunction. AB - INTRODUCTION: Biventricular cardiac pacemakers provide important hemodynamic benefit in selected patients with heart failure and severe left ventricular (LV) dysfunction. Nevertheless, these patients remain at high mortality risk. To address this issue, we examined mortality outcome in patients with heart failure treated with biventricular pacemakers alone and those treated with biventricular implantable cardioverter defibrillators (ICDs). METHODS AND RESULTS: The study population consisted of 126 consecutive patients with LV dysfunction and heart failure who received either a biventricular ICD (n = 62) or a biventricular pacemaker (n = 64) between January 1998 and December 2002. A minimum 12 months of follow-up was obtained in all survivors. ICD indications were conventional in all patients. Kaplan-Meier actuarial method and log rank statistics were used to calculate and compare survival rates in both groups. Comparison of mortality rates utilized Chi-square test. The two groups had similar clinical and demographic features, LV ejection fraction, and medication use. Average follow-up times were 13 +/- 11.8 months (range 4-60) and 18 +/- 13.2 months (range 0.5-53) for biventricular ICD and pacemaker groups, respectively. Overall mortality rate was significantly lower in the biventricular ICD group (13%, 8 deaths) compared to the pacemaker group (41%, 26 deaths) (P = 0.01). Further, the predominant survival benefit for ICD-treated patients becomes evident after the first 12 months of follow-up. CONCLUSION: The findings in this study, although necessarily limited in their interpretation by the absence of treatment randomization, suggest that biventricular ICDs offer a survival benefit compared to biventricular pacing alone. Furthermore, this benefit may be most apparent if other clinical factors do not preclude patient survival >1 year postimplant. PMID- 15333076 TI - Jumping in before the water is hot: the need to support randomized controlled clinical trials. PMID- 15333077 TI - Relationships between depolarization abnormality and repolarization abnormality in patients with Brugada syndrome: using body surface signal-averaged electrocardiography and body surface maps. AB - INTRODUCTION: Repolarization and depolarization abnormalities have been reported to be related to Brugada syndrome. METHODS AND RESULTS: We evaluated the relationships between repolarization abnormality and depolarization abnormality using 48-lead unipolar signal-averaged electrocardiograms and 87-lead unipolar body surface maps in 15 patients with Brugada-type ECGs. Data were compared with those from healthy control subjects (n = 5) and within subgroups of Brugada syndrome with (n = 8) and without (n = 7) ventricular arrhythmias (VA) induced by programmed electrical stimulation (PES). Eighty-seven-lead body surface maps were recorded, and potential maps were constructed to evaluate elevation of the ST segment 20 ms after the J point. Forty-eight-lead signal-averaged ECGs were recorded, and isochronal maps of duration of the delayed potential (dDP) were constructed to evaluate the dDP in each lead. Potential maps showed that patients with Brugada-type ECG, especially those with VA induced by programmed electrical stimulation, had greater elevation of the ST segment in the right ventricular outflow tract, especially at E5. Isochronal maps of dDP in the Brugada-type ECG group showed that maximum dDP was located at E5 and that the area with long dDP was larger than that in the control subjects. The dDPs at E7, E5, F7, and F5 in the VA-inducible group were significantly longer than those in the VA noninducible group. These results showed that the location of greater elevation in the ST segment coincided with the location of longer dDP. CONCLUSION: Repolarization abnormality and depolarization abnormality in the walls of both ventricles, especially in the right ventricular outflow tract, are related to the VA of Brugada syndrome. PMID- 15333078 TI - Sotalol reverses remodeled action potential in patients with chronic atrial fibrillation but does not prevent arrhythmia recurrence. AB - INTRODUCTION: Recurrence of atrial fibrillation (AF) may be related to AF-induced electrical remodeling characterized by shortening of the atrial action potential duration (APD) and loss of its rate adaptation. We investigated the effects of pretreatment with oral d,l-sotalol on rate-dependent changes in atrial monophasic action potential (MAP) duration after cardioversion of chronic AF with reference to the efficacy in preventing the arrhythmia recurrence. METHODS AND RESULTS: MAPs were recorded from the right atrium at six pacing cycle lengths (CLs) from 300 to 750 ms in 19 chronic AF patients after electrical cardioversion; 9 had been pretreated with oral d,l-sotalol (196 +/- 42 mg/day) for 7 days and 10 were untreated. MAP duration at 90% repolarization (MAPD90) in 11 control patients increased progressively with increases in CLs from 209 +/- 19 ms at CL = 300 ms to 264 +/- 28 ms at CL = 750 ms. In AF patients without sotalol, the CL-MAPD relation was shifted downward and flattened at longer CLs; MAPD90 values were 206 +/- 11 ms and 227 +/- 16 ms at CLs of 300 and 750 ms, respectively. MAPD90 values at CLs > or =500 ms in AF were significantly shorter than controls. In AF patients with sotalol, the normal CL-MAPD relation was preserved; MAPD90 increased from 226 +/- 19 ms to 282 +/- 46 ms in the CL range. AF recurred within 2 weeks after cardioversion in 14 of 24 patients pretreated with d,l-sotalol (216 +/- 51 mg/day) despite of continuation of sotalol treatment. CONCLUSION: Sotalol reverses AF-induced decrease in MAPD adaptation to rate in the atria of chronic AF patients, but this effect does not lead to prevention of AF recurrence. PMID- 15333079 TI - Is atrial remodeling a viable target for prevention of atrial fibrillation recurrence? PMID- 15333080 TI - Resolution of abnormal body surface maps in children with atrial septal defect after intracardiac repair. AB - INTRODUCTION: The genesis of repolarization abnormalities of ECG waveforms in atrial septal defect (ASD), which typically is characterized by right ventricular (RV) volume overload, has not been explored, particularly in association with postoperative hemodynamic improvement. The aim of this study was to evaluate the effects of reduced RV overload after ASD closure on depolarization and repolarization abnormalities on body surface maps (BSMs). METHODS AND RESULTS: BSMs of 14 children with ASD were recorded preoperatively and at early postoperative (1-6 months) and late postoperative (>9 months) stages. BSMs of 31 age-matched healthy children were studied as normal controls. Before intracardiac repair, QRS isopotential maps of children with ASD showed delayed RV breakthrough and subsequent rightward enlargement of the positive area with a maximum shifting to the right. Delayed conduction of the RV, particularly at the outflow tract area, was noted. The preoperative QRST isointegral maps exhibited the two-maximum pattern reflecting repolarization abnormality. The delayed appearance of breakthrough and delayed RV conduction on the QRS isopotential maps persisted from the preoperative to the late postoperative stage, whereas the two-maximum pattern on the QRST isointegral maps normalized to the one-dipole pattern at an early stage after repair. CONCLUSION: Abnormal repolarization parameters in ASD patients showed rapid improvement postoperatively, despite the persistence of depolarization abnormalities. Therefore, the two-maximum pattern on the QRST isointegral maps indicates a primary T wave change due to hemodynamic RV volume overload. PMID- 15333081 TI - Impact of biphasic electrical cardioversion of atrial fibrillation on early recurrent atrial fibrillation and shock efficacy. AB - INTRODUCTION: Early recurrent atrial fibrillation (ERAF) after external cardioversion of atrial fibrillation (AF) occurs in 12% to 26% of patients. Whether biphasic cardioversion has an impact on the incidence of ERAF after cardioversion of AF is unclear. METHODS AND RESULTS: Consecutive patients (n = 216, mean age 66 years, 71% male, 88% with structural cardiovascular disease or hypertension) underwent cardioversion with a biphasic (Bi) or monophasic (Mo) shock waveform in randomized fashion. Energies used were 120-150-200-200 Ws (Bi) or 200-300-360-360 Ws (Mo). The two study groups (Bi vs Mo) did not differ with regard to age, sex, body mass index, underlying cardiovascular disease, left atrial diameter, left ventricular ejection fraction, duration of AF fibrillation, and antiarrhythmic drug therapy. Mean delivered energy was significantly lower in the Bi group (Bi: 186 +/- 143 Ws vs Mo: 324 +/- 227 Ws; P < 0.001). Overall incidence of ERAF (AF relapse within 1 minute after successful cardioversion) was 8.9% and showed no difference between the two groups (Bi: 8.1% vs Mo: 9.7%, P = NS). Cardioversion was successful in 95.4% of patients. The success rate was comparable in both groups (Bi: 94.3% vs Mo 96.8%; P = NS). First shock efficacy did not differ between Bi and Mo (76.4% vs 67.7%; P = NS). Mean number of shocks were 1.4 shocks per patient in both groups. CONCLUSION: Biphasic cardioversion allows comparable success rates with significantly lower energies. However, the incidence of ERAF is not influenced by biphasic cardioversion. With the energies used, biphasic and monophasic shock waveforms are comparable with regard to first shock and cumulative shock efficacy. PMID- 15333082 TI - Early recurrence of atrial fibrillation: does the shock waveform matter? PMID- 15333083 TI - Myocardial ischemia and induction of sustained ventricular tachyarrhythmias: evaluation using dobutamine stress echocardiography-electrophysiologic testing. AB - INTRODUCTION: This study examined the relationship between dobutamine facilitation of ventricular tachyarrhythmia (VT) inducibility with programmed electrical stimulation (PES) and dobutamine stress-induced myocardial ischemia. METHODS AND RESULTS: Twenty patients with prior myocardial infarction and cardiac arrest or sustained VT but no sustained VT induced at baseline electrophysiologic testing underwent repeat PES during dobutamine infusion. Ischemia (new or worsened wall-motion abnormality) was documented by echocardiography performed in conjunction with PES. Eight patients were receiving Class I or III antiarrhythmic drugs and seven beta-blockers. Dobutamine facilitated induction of sustained VT in 16 patients (80%) and provoked ischemia in 13 patients (65%). Induction of VT was associated with ischemia in 9 patients (56%). VTs associated with ischemia were induced at higher dobutamine doses (26 +/- 11 vs 11 +/- 10 microg/kg per min, P = 0.02) than were VTs without ischemia (n = 7). Among 13 patients with provoked ischemia, 9 (69%) had VTs induced and 4 remained noninducible. The onset of ischemia occurred at the same dose as induction of VT in 5 patients and at a lower dose in 4 patients. Monomorphic VT (318 +/- 59 ms) was induced in 13 patients, of whom 8 (62%) had ischemia. The ECG morphology of VT suggested an origin in a myocardial segment that demonstrated initial viability at low doses then ischemic dysfunction at higher doses preceding VT induction in 7 (88%) of 8 patients. CONCLUSION: Dobutamine enhances inducibility of sustained VTs during PES. The temporal and anatomic association of dobutamine-induced ischemia and VT suggests that at high dobutamine doses, ischemia may contribute to ventricular arrhythmia inducibility in some patients. PMID- 15333084 TI - Mechanisms of ventricular tachycardia induction. PMID- 15333085 TI - Atrial fibrillatory wave characteristics on surface electrogram: ECG to ECG repeatability over twenty-four hours in clinically stable patients. AB - INTRODUCTION: Fibrillatory waves on the surface ECG have been scrutinized to allow inferences about underlying mechanisms and pathophysiology, based on the premise that fibrillatory waves do not vary "randomly" but provide a consistent reflection of the underlying state of the atria in an individual patient. This premise is untested. METHODS AND RESULTS: Ten standard ECGs were recorded over a 24-hour period in each of 20 clinically stable inpatients with atrial fibrillation. After QRS-T cancellation, the remainder fibrillatory waves were analyzed. Interpatient versus intrapatient differences in fibrillatory wave characteristics were evaluated by analysis of variance (ANOVA). The fibrillatory wave peak-to-peak amplitude of all the patients ranged from 0.06 to 0.35 mV, whereas 1 SD of the amplitude for each patient ranged from 0.004 to 0.053 mV. Short-term peak frequencies of all the patients ranged from 4.6 to 8.0 Hz, whereas 1 SD for each patient varied from 0.2 to 0.5 Hz. For these and all other parameters tested, interpatient differences were significantly greater compared to intrapatient differences (P < 0.0001). CONCLUSION: Fibrillatory wave characteristics are repeatable from ECG to ECG over 24 hours for clinically stable patients, whereas substantial differences are present between patients. Further study of the relationship of such characteristics to pathophysiology and management decisions is valid and warranted. PMID- 15333086 TI - Is analysis of fibrillatory waves useful for treatment of atrial fibrillation? PMID- 15333087 TI - Prevalence of asymptomatic recurrences of atrial fibrillation after successful radiofrequency catheter ablation. AB - INTRODUCTION: The long-term efficacy of radiofrequency catheter ablation of atrial fibrillation (AF) has been based on patient-reported symptoms suggestive of AF. However, asymptomatic recurrences of AF may remain undetected. The aim of this study was to determine the prevalence of asymptomatic recurrences of AF after an apparently successful catheter ablation procedure for AF. METHODS AND RESULTS: Among 244 consecutive patients (mean age 53 +/- 11 years) who underwent a pulmonary vein isolation procedure for symptomatic paroxysmal AF and who reported no symptoms of recurrent AF at > or =6 months after the procedure, 60 patients with a history of > or =1 episode of AF per week were asked to participate in this study. Preablation, these patients had experienced 19 +/- 13 episodes of AF per month. The patients were provided with a patient-activated transtelephonic event recorder for 30 days, a mean of 642 +/- 195 days after the ablation procedure, and were asked to record and transmit recordings on a daily basis and whenever they felt palpitations. Seven patients (12%) felt palpitations during the study, although they had not experienced symptoms previously. Each of these 7 patients had an episode of AF documented with the event monitor during symptoms. In these 7 patients, the mean number of episodes per month decreased from 19 +/- 14 preablation to 3 +/- 1 postablation (P < 0.001). Among the 53 asymptomatic patients, an episode of AF was captured in 1 (2%) patient during the study period. CONCLUSION: Asymptomatic recurrences of AF after an apparently successful catheter ablation procedure for symptomatic paroxysmal AF are infrequent. PMID- 15333088 TI - Asymptomatic atrial fibrillation in symptomatic patients. PMID- 15333089 TI - Atrial natriuretic peptides during experimental atrial tachycardia: role of developing tachycardiomyopathy. AB - INTRODUCTION: Atrial tachycardia and chronic heart failure (CHF) are associated with elevated levels of atrial natriuretic peptide (ANP) and its amino terminal part NT-ANP. Chronic high atrial rates may cause CHF due to a rapid ventricular response. The aim of this study was to establish the contribution of elevated atrial rate and of high ventricular rate, resulting in CHF, on ANP and NT-ANP levels during chronic atrial tachycardia. METHODS AND RESULTS: Thirteen goats (AV paced group) were subjected to 4 weeks of rapid AV pacing with an atrial and ventricular rate of 240 beats/min. Another five goats (A-paced group) were subjected to 4 weeks of atrial pacing at 240 beats/min while the ventricular rate was kept low and regular at 80 beats/min. Pacing was interrupted only for measurement of right atrial (RA) and left ventricular (LV) diameter and sampling for ANP, NT-ANP, and renin. In the AV-paced group, RA and LV diameter reached 152% and 109% of baseline values, respectively. Both ANP and NT-ANP (8.3 +/- 9.2 pmol/L and 0.5 +/- 0.4 nmol/L at baseline, respectively) increased progressively (53.1 +/- 37.9 pmol/L and 2.0 +/- 0.9 nmol/L, respectively, after 4 weeks). There was a significant correlation between the magnitude of atrial dilation and natriuretic peptide levels after 3 days. In A-paced goats, however, RA and LV diameters did not change. Furthermore, ANP and NT-ANP levels (9.1 +/- 6.0 pmol/L and 0.8 +/- 0.2 nmol/L at baseline, respectively) were unchanged after 4 weeks (5.3 +/- 3.4 pmol/L and 0.6 +/- 0.2 nmol/L, respectively). CONCLUSION: Elevated levels of ANPs during chronic atrial tachycardia are related to a high ventricular rate rather than a high atrial rate alone. Rather than atrial tachycardia, the atrial hemodynamic burden is an important determinant of the sustained ANP response. PMID- 15333090 TI - Atrial tachycardia and congestive heart failure: new insights from an old experimental model. PMID- 15333091 TI - Canine model of Brugada syndrome using regional epicardial cooling of the right ventricular outflow tract. AB - INTRODUCTION: Myocardial cooling can induce J point elevation (Osborn wave) as seen on ECG of the Brugada syndrome by activating transient outward current (Ito) and causing a spike-and-dome configuration of the monophasic action potential (MAP) in the ventricular epicardium in isolated canine ventricular wedge preparations. We determined the effect of regional epicardial cooling of the right ventricular outflow tract (RVOT) on surface ECG and ventricular vulnerability in the dog. METHODS AND RESULTS: In 12 dogs, a cooling device (20 mm diameter) was attached to the RVOT epicardium, and surface ECG, epicardial MAP, and endocardial MAP were recorded. Regional cooling (29.7 degrees C +/- 2.2 degrees C) elevated the J point from 0.05 +/- 0.06 mV to 0.12 +/- 0.11 mV and induced T wave inversion (from 0.02 +/- 0.12 mV to -0.27 +/- 0.20 mV) in lead V1 in association with "spike-and-dome" configuration of the epicardial MAP. Cooling prolonged MAP duration in the RVOT epicardium from 172 +/- 27 ms to 213 +/- 30 ms (P < 0.01) but not in the RV endocardium and increased transmural dispersion of MAP duration from 9 +/- 8 ms to 44 +/- 21 ms (P < 0.01). Cooling also prolonged the QT interval in lead V1 from 191 +/- 19 ms to 212 +/- 23 ms (P < 0.05), but not in lead V5, and increased spatial dispersion of QT interval from 7 +/- 5 ms to 20 +/- 10 ms (P < 0.01). QT interval in lead V1 correlated positively with MAP duration in the RVOT epicardium (r = 0.75). T wave amplitude in lead V1 correlated inversely with transmural dispersion of MAP duration in the RVOT (r = 0.74). Vagal nerve stimulation accentuated the cooling-induced changes. During cooling, ventricular fibrillation was induced by a single extrastimulus in 2 of 4 dogs, and additional vagal nerve stimulation during isoproterenol administration induced spontaneous ventricular fibrillation in one dog. CONCLUSION: Localized epicardial cooling of the RVOT could be an in vivo experimental model of Brugada syndrome. PMID- 15333092 TI - Link between hypothermia and the Brugada syndrome. PMID- 15333093 TI - Necessity for biatrial ablation to achieve bidirectional cavotricuspid isthmus conduction block in a patient following senning operation. AB - We report the case of a 28-year-old male patient with a 17-year history of recurrent symptomatic atrial tachyarrhythmia following Senning operation for transposition of the great arteries. Biatrial electroanatomic mapping and entrainment mapping revealed counterclockwise peri-tricuspid annulus reentry in which cavotricuspid isthmus tissue in both systemic and pulmonary venous atria was involved. Linear ablation of the cavotricuspid isthmus in the pulmonary venous atrium terminated the tachycardia but did not block the isthmus conduction, and the tachycardia was reinduced. Bidirectional isthmus conduction block could be achieved only after additional linear ablation targeting the cavotricuspid isthmus tissue in the systemic venous atrium. We conclude that biatrial ablation may be necessary in order to achieve bidirectional isthmus block and prevent tachycardia recurrence in some patients following Senning or Mustard operation. PMID- 15333094 TI - Superior vena cava rhythm masquerading as normal sinus rhythm. AB - We report the case of a patient with persistent cardiac rhythm originating from the superior vena cava (3 cm above the vena cava-atrial junction). It was detected by noncontact balloon mapping before induction of tachycardia and confirmed by conventional contact mapping with image studies. Thus, a 12-lead ECG showing normal morphologies of P waves may not indicate that the P waves are of sinus node origin. PMID- 15333095 TI - Death due to an implantable cardioverter defibrillator. AB - Inappropriate therapy due to noise oversensing caused a true ventricular fibrillation (VF) and death of a patient. A 49-year-old patient with a history of dilated cardiomyopathy received a double-chamber implantable cardioverter defibrillator (ICD) in 1991 for a sustained inducible ventricular tachycardia (VT). One appropriate shock delivered in 1994 terminated an episode of VT. The generator was replaced in 1995 and in 2000, and was connected to the initial leads. Three months after the second replacement, the patient received six consecutive shocks related to detection of noise interpreted as VF. Unfortunately, the sixth shock triggered a true VF, which was not treated due to end of the therapeutic sequence, and the patient died. The causes of the dysfunction are discussed. PMID- 15333096 TI - Radiofrequency ablation of idiopathic ventricular fibrillation guided by noncontact mapping. AB - A 32-year-old man with idiopathic ventricular fibrillation and an implantable cardioverter defibrillator presented during a ventricular fibrillation storm. Frequent monomorphic ventricular ectopics with left bundle branch block morphology were documented, some of which initiated fibrillation. He underwent noncontact mapping of the right ventricle, during which the ventricular ectopics were mapped to a site in the free wall displaying a diastolic potential 80 ms before ectopic QRS onset. Following three radiofrequency energy applications, the ectopics were abolished. After 11-month follow-up, he has experienced no further arrhythmias. Noncontact mapping may identify ablatable triggers of ventricular fibrillation and lead to successful outcomes even when only single ectopics are present. PMID- 15333097 TI - Left atrial-esophageal fistula following radiofrequency catheter ablation of atrial fibrillation. AB - This report describes a fatal case of left atrial-esophageal fistula occurring in a 72-year-old man after a radiofrequency catheter ablation of paroxysmal atrial fibrillation. Catheter ablation was performed around the pulmonary vein using an 8-mm-tip electrode (60 W or 55 degrees C) guided by a 25-mm circular catheter. On day 22 of follow-up, the patient presented with seizures followed by hematemesis due to left atrial-esophageal fistula. His clinical condition deteriorated, and he died of speticemia. Thus, left atrial-esophageal fistula is a sever complication of radiofrequency catheter ablation of the left atrial posterior wall. PMID- 15333098 TI - Asynchronous ventricular pacing triggering ventricular fibrillation. PMID- 15333099 TI - Narrow QRS complex tachycardia following tricuspid valve replacement. PMID- 15333100 TI - Utility of a nonfluoroscopic navigation system for pulmonary vein isolation. PMID- 15333101 TI - Novel method to integrate three-dimensional computed tomographic images of the left atrium with real-time electroanatomic mapping. PMID- 15333102 TI - Pulmonary vein narrowing following atrial fibrillation ablation. PMID- 15333106 TI - From the editors: external peer review at HSR. PMID- 15333107 TI - Physician workforce data: when the best is not good enough. PMID- 15333108 TI - Potentially inappropriate medication prescriptions among elderly nursing home residents: their scope and associated resident and facility characteristics. AB - OBJECTIVE: To estimate the scope of potentially inappropriate medication prescriptions (PIRx) among elderly residents in U.S. nursing homes (NHs), and to examine associated resident and facility characteristics. DATA SOURCES: The 1996 Medical Expenditure Panel Survey Nursing Home Component (MEPS NHC), a survey of a nationally representative sample of NHs and residents. STUDY DESIGN: The PIRx, defined by Beers's consensus criteria (1991, 1997), was identified using up to a year's worth of NH prescribed medicine data for each resident. The study sample represented 1.6 million NH residents (n=3,372). RESULTS: At a minimum, 50 percent of all residents aged 65 or older, with an NH stay of three months or longer received at least one PIRx in 1996. The most common PIRx involved propoxyphene, diphenhydramine, hydroxyzine, oxybutynin, amitriptyline, cyproheptadine, iron supplements, and ranitidine. Resident factors associated with greater odds of PIRx were Medicaid coverage, no high school diploma, and nondementia mental disorders. Facility factors were more beds and lower RN-to-resident ratio. Factors associated with lower odds of PIRx were fewer medications, residents with communication problems, and being in an accredited NH. Onsite availability of pharmacists or mental health providers was not related. IMPLICATIONS: With quality of care and patient safety as major public health concerns, effective policies are needed to avoid PIRx occurrences and improve the quality of prescribing among elderly residents in NHs. Additional studies are needed to determine the impact of PIRx on this NH population. PMID- 15333109 TI - Socioeconomic disparities in the use of home health services in a medicare managed care population. AB - OBJECTIVE: To investigate socioeconomic disparities in access to home health visits and durable medical equipment by persons enrolled in two Medicare managed care health plans. DATA SOURCES: A telephone survey of 4,613 Medicare managed care enrollees conducted between April and October of 2000 and linked to administrative claims for a subsequent 12-month period. STUDY DESIGN: We estimated a series of logistic regression models to determine which socioeconomic factors were related to home health visits and the use of durable medical equipment (DME) among Medicare managed care enrollees. PRINCIPAL FINDINGS: Controlling for health and demographic differences, Medicare managed care enrollees in the lowest tertile for nonhousing assets had 50 percent greater odds than those in the highest tertile of having one or more home health visits. All else equal, enrollees with less than a high school education had 30 percent lower odds than those who had graduated from high school of using durable medical equipment. CONCLUSIONS: Medicare managed care enrollees of low socioeconomic status do not appear to have reduced access to home health visits; however, use of durable medical equipment is considerably lower for enrollees with less than a high school education. Physicians and therapists working with Medicare managed care enrollees may want to actively target DME prescriptions to those with educational disadvantages. PMID- 15333110 TI - Physical rehabilitation following medicare prospective payment for skilled nursing facilities. AB - OBJECTIVE: To examine the effect of the Medicare prospective payment system (PPS) for skilled nursing facilities (SNF) on the delivery of rehabilitation therapy to residents. DATA SOURCES: Resident-level data are based on the Resident Assessment Instrument Minimum Data Set for nursing facilities. All elderly residents admitted to SNFs in Michigan and Ohio in 1998 and 1999 form the study population (n=99,952). STUDY DESIGN: A differences-in-differences identification strategy is used to compare rehabilitation therapy for SNF residents before and after a change in Medicare SNF payment. Logistic and linear regression analyses are used to examine the effect of PPS on receipt of physical, occupational, or speech therapy and total therapy time. DATA EXTRACTION: Data for the present study were extracted from the University of Michigan Assessment Archive Project (UMAAP). One assessment was obtained for each resident admitted to nursing facilities during the study period. PRINCIPAL FINDINGS: The introduction of PPS for all U.S. Medicare residents in July of 1998 was associated with specific targeting of rehabilitation treatment time to the most profitable levels of therapy. The PPS was also associated with increased likelihood of therapy but less rehabilitation therapy time for Medicare residents. CONCLUSIONS: The present results indicate that rehabilitation therapy is sensitive to the specific payment incentives associated with PPS. PMID- 15333111 TI - Mental illness and length of inpatient stay for medicaid recipients with AIDS. AB - OBJECTIVE: To examine the associations between comorbid mental illness and length of hospital stays (LOS) among Medicaid beneficiaries with AIDS. DATA SOURCE AND COLLECTION/STUDY SETTING: Merged 1992-1998 Medicaid claims and AIDS surveillance data obtained from the State of New Jersey for adults with >or=1 inpatient stay after an AIDS diagnosis from 1992 to 1996. STUDY DESIGN: Observational study of 6,247 AIDS patients with 24,975 inpatient visits. Severe mental illness (SMI) and other less severe mental illness (OMI) diagnoses at visits were ascertained from ICD-9 Codes. About 4 percent of visits had an SMI diagnosis; 5 percent had an OMI diagnosis; 43 percent did not have a mental illness diagnosis, but were patients who had been identified as having an SMI or OMI history; and 48 percent were from patients with no identified history of mental illness. PRINCIPAL FINDINGS: The overall mean hospital LOS was 12.7 days. After adjusting for measures of HIV disease severity and health care access in multivariate models, patients presenting with primary and secondary severe mental illness (SMI) diagnoses had approximately 32 percent and approximately 11 percent longer LOS, respectively, than did similar patients without a mental illness history (p<0.001 for each). But in these adjusted models of length of stay: (1) diagnosis of OMI was not related to LOS, and (2) in the absence of a mental illness diagnosed at the visit, an identified history of either SMI or OMI was also not related to LOS. In adjusted models of time to readmission for a new visit, current diagnosis of SMI or OMI and in the absences of a current diagnosis, history of SMI or OMI all tended to be associated with quicker readmission. CONCLUSIONS: This study finds greater (adjusted) LOS for AIDS patients diagnosed with severe mental illness (but not for those diagnosed with less severe mental comorbidity) at a visit. The effect of acute severe mental illness on hospitalization time may be comparable to that of an acute AIDS opportunistic illness. While previous research raises concerns that mental illness increases LOS by interfering with treatment of HIV conditions, the associations here may simply indicate that extra time is needed to treat severe mental illnesses or arrange for discharge of afflicted patients. PMID- 15333112 TI - The cost-effectiveness of independent housing for the chronically mentally ill: do housing and neighborhood features matter? AB - OBJECTIVE: To determine the effects of housing and neighborhood features on residential instability and the costs of mental health services for individuals with chronic mental illness (CMI). DATA SOURCES: Medicaid and service provider data on the mental health service utilization of 670 individuals with CMI between 1988 and 1993 were combined with primary data on housing attributes and costs, as well as census data on neighborhood characteristics. Study participants were living in independent housing units developed under the Robert Wood Johnson Foundation Program on Chronic Mental Illness in four of nine demonstration cities between 1988 and 1993. STUDY DESIGN: Participants were assigned on a first-come, first-served basis to housing units as they became available for occupancy after renovation by the housing providers. Multivariate statistical models are used to examine the relationship between features of the residential environment and three outcomes that were measured during the participant's occupancy in a study property: residential instability, community-based service costs, and hospital based service costs. To assess cost-effectiveness, the mental health care cost savings associated with some residential features are compared with the cost of providing housing with these features. DATA COLLECTION/EXTRACTION METHODS: Health service utilization data were obtained from Medicaid and from state and local departments of mental health. Non-mental-health services, substance abuse services, and pharmaceuticals were screened out. PRINCIPAL FINDINGS: Study participants living in newer and properly maintained buildings had lower mental health care costs and residential instability. Buildings with a richer set of amenity features, neighborhoods with no outward signs of physical deterioration, and neighborhoods with newer housing stock were also associated with reduced mental health care costs. Study participants were more residentially stable in buildings with fewer units and where a greater proportion of tenants were other individuals with CMI. Mental health care costs and residential instability tend to be reduced in neighborhoods with many nonresidential land uses and a higher proportion of renters. Mixed-race neighborhoods are associated with reduced probability of mental health hospitalization, but they also are associated with much higher hospitalization costs if hospitalized. The degree of income mixing in the neighborhood has no effect. CONCLUSIONS: Several of the key findings are consistent with theoretical expectations that higher-quality housing and neighborhoods lead to better mental health outcomes among individuals with CMI. The mental health care cost savings associated with these favorable features far outweigh the costs of developing and operating properties with them. Support for the hypothesis that "diverse-disorganized" neighborhoods are more accepting of individuals with CMI and, hence, associated with better mental health outcomes, is mixed. PMID- 15333113 TI - The effects of state mental health parity legislation on perceived quality of insurance coverage, perceived access to care, and use of mental health specialty care. AB - OBJECTIVE: To assess the impacts of recent state mental health parity legislation on perceived quality of health insurance coverage, perceived access to needed health care, and use of mental health specialty services by individuals with likely need for mental health care. DATA SOURCES: The study sample came from two waves of a national household survey first fielded in 1997-1998 and then in 2000 2001. The analysis used a subset of the sample. STUDY DESIGN: The study took the Difference-in-Difference-in-Difference approach to investigate changes in self perceived quality of health insurance coverage and access to needed health care, and use of mental health specialty care by the group with mental disorders (relative to those without) in states with parity legislation of different comprehensiveness (relative to the nonparity states) in the years after the law (relative to before the law). PRINCIPAL FINDINGS: Overall, there were no significant or consistent effects of the parity legislation. Descriptive statistics showed significant changes in some (but not all) outcome variables, but these results disappeared in detailed statistical analyses by controlling for important covariates. CONCLUSIONS: The null findings of the effects of state mental health parity mandates suggest that under ERISA (Employee Retirement Income Security Act), the scope of state parity legislation may have been restricted because of large proportion of self-insured employers. Furthermore, comprehensiveness of state legislation appears to be related to the traditional level of use of mental health specialty care, which becomes another confounder for the potential policy effects. PMID- 15333114 TI - Cost shifting to jails after a change to managed mental health care. AB - OBJECTIVE: To determine whether managed mental health care for Medicaid enrollees in King County, Washington, has led to indirect cost-shifting to substitute treatments, such as jails and state mental hospitals that are free goods to providers. DATA SOURCES: Complete service records for 47,300 adults who used at least one of the following systems from 1993 to 1998: King County jail system, Medicaid, or the King County mental health system. Data were also obtained from the Washington State Hospital System. STUDY DESIGN: A quasi-experimental analysis that compares the difference in outcomes between the pre- and post-managed care periods for Medicaid enrollees compared to non-Medicaid enrollees. The outcomes jail costs, state hospital costs, and county outpatient mental health costs-were estimated with two-part difference-in-differences models. The regressions control for person-level fixed effects on up to 66 months of data per person. DATA COLLECTION METHODS: Administrative data were collected from the jail, Medicaid, and mental health systems, then merged and cleaned. Additional data on costs were obtained in interviews. PRINCIPAL FINDINGS: There is a striking increase in the probability of jail use for persons on Medicaid following the introduction of managed care. There was a significant decrease in expenditures in the county mental health system for outpatient care. CONCLUSIONS: Managed care led to indirect cost-shifting, probably through poor access to services, which may have led to an increased probability of jail detention. PMID- 15333115 TI - Associations of race, education, and patterns of preventive service use with stage of cancer at time of diagnosis. AB - OBJECTIVE: To go beyond the documentation of disparities by race and SES by analyzing health behaviors regarding preventive and cancer screening services and determining if these behaviors are associated with stage of cancer when first diagnosed. DATA: Stage of cancer for Medicare patients diagnosed in 1995 with breast, colorectal, uterine, ovarian, prostate, bladder, or stomach cancer; and use of influenza and pneumonia immunization, mammography, pap smear, colon cancer screening, and the prostate specific antigen test during the two years preceding diagnosis of cancer. STUDY DESIGN: Hypothesis tested: health behaviors regarding use of preventive and cancer screening services are associated with stage of cancer when first diagnosed. DATA COLLECTION/EXTRACTION METHODS: Information was extracted from the database formed by the linkage of Surveillance, Epidemiology, and End Results (SEER) cancer registries with Medicare files. PRINCIPAL FINDINGS: Black and white patients (of higher and lower SES) who used more of the preventive and cancer screening services were at a lower risk of having late stage cancer for six cancers studied (breast, colorectal [male and female], prostate, uterine, and male bladder cancer) than their counterparts who used fewer of these services. CONCLUSIONS: The use of preventive and cancer screening services is a health behavior associated with better health outcomes for the elderly diagnosed with cancer. The lack of preventive service use can serve as a marker for identifying persons at risk of late stage cancer when first diagnosed. Strategies that encourage the use of preventive services by low users of these services are likely to reinforce a range of healthy behaviors that help to ameliorate disparities in health outcomes. PMID- 15333116 TI - Monitoring visual status: why patients do or do not comply with practice guidelines. AB - OBJECTIVE: To determine factors affecting compliance with guidelines for annual eye examinations for persons diagnosed with diabetes mellitus (DM) or age-related macular degeneration (ARMD). DATA SOURCES/STUDY SETTING: Nationally representative, longitudinal sample of individuals 65+ drawn from the National Long-Term Care Survey (NLTCS) with linked Medicare claims records from 1991 to 1999. STUDY DESIGN: Medicare beneficiaries were followed from 1991 to 1999, unless mortality intervened. All claims data were analyzed for presence of ICD-9 codes indicating diagnosis of DM or ARMD and the performance of eye exams. The dependent variable was a binary indicator for whether a person had an eye exam or not during a 15-month period. Independent variables for demographics, living conditions, supplemental insurance, income, and other factors affecting the marginal cost and benefit of an eye exam were assessed to determine reasons for noncompliance. DATA COLLECTION/EXTRACTION METHODS: Panel data were created from claims files, 1991-1999, merged with data from the NLTCS. PRINCIPAL FINDINGS: The probability of having an exam reflected perceived benefits, which vary by patient characteristics (e.g., education, no dementia), and factors associated with the ease of visit. African Americans were much less likely to be examined than were whites. CONCLUSIONS: Having an exam reflects multiple factors. However, much of the variation in the probability of an exam remained unexplained as were reasons for the racial differences in use. PMID- 15333118 TI - Plan, geographical, and temporal variation of consumer assessments of ambulatory health care. AB - OBJECTIVE: To quantify contributions of health plans and geography to variation in consumer assessments of health plan quality. DATA SOURCES: Responses of beneficiaries of Medicare managed care plans to the Consumer Assessment of Health Plans Study (CAHPS(R)) survey. Our data included more than 700,000 survey responses assessing 381 Medicare managed care (MMC) contracts over a period of five years. STUDY DESIGN: The survey was administered to a nationally representative sample of beneficiaries of Medicare managed care plans. PRINCIPAL FINDINGS: Member assessments of their health plans, customer service functions, and prescription drug benefits varied most across health plans; these also varied the most over time. Assessments of direct interactions with doctors and their practices were more affected by geographical location, and these assessments were quite stable over time. A health plan's global rating often changed significantly between consecutive years, but only rarely were there such changes in ratings of care or doctor. Nationally, mean assessments tended to decrease over the study period. CONCLUSIONS: Our findings suggest that ratings of plans and reports about customer service and prescription access are affected by plan policies, benefits design, and administrative structures that can be changed relatively quickly. Conversely, assessments of other aspects of care are largely determined by characteristics of provider networks that are relatively stable. A consumer survey is unlikely to detect meaningful changes in quality of care from year to year unless quality improvement measures are developed that have substantially larger effects, possibly through area-wide initiatives, than historical temporal variations in quality. PMID- 15333117 TI - Posthospital care transitions: patterns, complications, and risk identification. AB - OBJECTIVES: To (1) describe patterns of posthospital care transitions; (2) characterize these patterns as uncomplicated or complicated; (3) identify those at greatest risk for complicated transitions. DATA SOURCES/STUDY SETTING: The Medicare Current Beneficiary Survey was used to identify beneficiaries aged 65 and older who were discharged from an acute care hospital in 1997-1998. STUDY DESIGN: Patterns of posthospital transfers were described over a 30-day time period following initial hospital discharge. Uncomplicated posthospital care patterns were defined as a sequence of transfers from higher-to lower-intensity care environments without recidivism, while complicated posthospital care patterns were defined as the opposite sequence of events. Indices were developed to identify patients at risk for complicated transitions. PRINCIPAL FINDINGS: Forty-six distinct types of care patterns were observed during the 30 days following hospital discharge. Among these patterns, 444 episodes (61.2 percent) were limited to a single transfer, 130 episodes (17.9 percent) included two transfers, 62 episodes (8.5 percent) involved three transfers, and 31 episodes (4.3 percent) involved four or more transfers. Fifty-nine episodes (8.1 percent) resulted in death. Between 13.4 percent and 25.0 percent of posthospital care patterns in the 1998 sample were classified as complicated. The area under the receiver operating curve was 0.771 for a predictive index that utilized administrative data and 0.833 for an index that used a combination of administrative and self-reported data. CONCLUSIONS: Posthospital care transitions are common among Medicare beneficiaries and patterns of care vary greatly. A significant number of beneficiaries experienced complicated care transitions-a finding that has important implications for both patient safety and cost containment efforts. Patients at risk for complicated care patterns can be identified using data available at the time of hospital discharge. PMID- 15333119 TI - Communities and hospitals: social capital, community accountability, and service provision in U.S. community hospitals. AB - OBJECTIVES: The study related community social capital to the level of community accountability and provision of community-oriented services in U.S. community hospitals. STUDY SETTING: The sample included 1,383 community hospitals that participated in the 1997 American Hospital Association's (AHA) Hospital Annual and Governance Surveys. DATA SOURCES: (1) The 1997 AHA Annual Hospital Survey, (2) the 1997 AHA Hospital Governance Survey, (3) the DDB Needham Market Facts Survey, (4) the 1996 County Election Data File, and (5) the 1998 Area Resource File. RESEARCH DESIGN: The study used a mix of longitudinal and cross-sectional data. KEY FINDINGS: We identified two distinct indicators of social capital community participation and voting participation. Community accountability in hospitals was unrelated to either indicator. Hospitals' provision of community oriented health services was negatively associated with community participation but unrelated with voting participation. The interaction between voting participation and community representation on hospital governance was positively associated with community accountability and provision of community-oriented health services. CONCLUSION: Neither community participation nor voting participation was sufficient to influence hospital behavior. The positive finding associated with the interaction between voting participation and community representation on hospital governance underscored the importance of an active political culture in influencing hospital behavior, without which the installation of community representatives on hospital governance might be more symbolic than actually serving the health concerns of community residents. PMID- 15333120 TI - The unintended impact of welfare reform on the medicaid enrollment of eligible immigrants. AB - BACKGROUND: During welfare reform, Congress passed legislation barring legal immigrants who entered the United States after August 1996 from Medicaid for five years after immigration. This legislation intended to bar only new immigrants (post-1996 immigrants) from Medicaid. However it may have also deterred the enrollment of legal immigrants who immigrated before 1996 (pre-1996 immigrants) and who should have remained Medicaid eligible. OBJECTIVES: To compare the Medicaid enrollment of U.S.-born citizens to pre-1996 immigrants, before and after welfare reform, and to determine if variation in state Medicaid policies toward post-1996 immigrants modified the effects of welfare reform on pre-1996 immigrants. DATA SOURCE/STUDY DESIGN: Secondary database analysis of cross sectional data from 1994-2001 of the U.S. Census Bureau, Annual Demographic Survey of March Supplement of the Current Population Survey. SUBJECTS: Low income, U.S.-born adults (N=116,307) and low-income pre-1996 immigrants (N=24,367) before and after welfare reform. MEASURES: Self-reported Medicaid enrollment. RESULTS: Before welfare reform, pre-1996 immigrants were less likely to enroll in Medicaid than the U.S.-born (OR=0.55; 95 percent CI, 0.51-0.59). After welfare reform, pre-1996 immigrants were even less likely to enroll in Medicaid. The proportion of immigrants in Medicaid dropped 3 percentage points after 1996; for the U.S.-born it dropped 1.6 percentage points (p=0.012). Except for California, state variation in Medicaid policy toward post-1996 immigrants did modify the effect of welfare reform on pre-1996 immigrants. CONCLUSIONS: Federal laws limiting the Medicaid eligibility of specific subgroups of immigrants appear to have had unintended consequences on Medicaid enrollment in the larger, still eligible immigrant community. Inclusive state policies may overcome this effect. PMID- 15333121 TI - Availability of safety net providers and access to care of uninsured persons. AB - OBJECTIVE: To understand how proximity to safety net clinics and hospitals affects a variety of measures of access to care and service use by uninsured persons. DATA SOURCES: The 1998-1999 Community Tracking Study household survey, administered primarily by telephone survey to households in 60 randomly selected communities, linked to data on community health centers, other free clinics, and safety net hospitals. STUDY DESIGN: Instrumental variable estimation of multivariate regression models of several measures of access to care (having a usual source of care, unmet or delayed medical care needs, ambulatory service use, and overnight hospital stays) against endogenous measures of distances to the nearest community health center and safety net hospital, controlling for characteristics of uninsured persons and other area characteristics that are related to access to care. The models are estimated with data from a nationally representative sample of uninsured people. PRINCIPAL FINDINGS: Shorter distances to the nearest safety net providers increase access to care for uninsured persons. Failure to account for the endogeneity of distance to safety net providers on access to care generally leads to finding little or no safety net effects on access. CONCLUSIONS: Closer proximity to the safety net increases access to care for uninsured persons. However, the improvements in access to care are relatively small compared with similar measures of access to care for insured persons. Modest expansion of the safety net is unlikely to provide a full substitute for insurance coverage expansions. PMID- 15333122 TI - Subsidies and the demand for individual health insurance in California. AB - OBJECTIVE: To estimate the effect of changes in premiums for individual insurance on decisions to purchase individual insurance and how this price response varies among subgroups of the population. DATA SOURCE: Survey responses from the Current Population Survey (http://www.bls.census.gov/cps/cpsmain.htm), the Survey of Income and Program Participation (http://www.sipp.census.gov/sipp), the National Health Interview Survey (http://www.cdc.gov/nchs/nhis.htm), and data about premiums and plans offered in the individual insurance market in California, 1996 2001. STUDY DESIGN: A logit model was used to estimate the decisions to purchase individual insurance by families without access to group insurance. This was modeled as a function of premiums, controlling for family characteristics and other characteristics of the market. A multinomial model was used to estimate the choice between group coverage, individual coverage, and remaining uninsured for workers offered group coverage as a function of premiums for individual insurance and out-of-pocket costs of group coverage. PRINCIPAL FINDINGS: The elasticity of demand for individual insurance by those without access to group insurance is about -.2 to -.4, as has been found in earlier studies. However, there are substantial differences in price responses among subgroups with low-income, young, and self-employed families showing the greatest response. Among workers offered group insurance, a decrease in individual premiums has very small effects on the choice to purchase individual coverage versus group coverage. CONCLUSIONS: Subsidy programs may make insurance more affordable for some families, but even sizeable subsidies are unlikely to solve the problem of the uninsured. We do not find evidence that subsidies to individual insurance will produce an unraveling of the employer-based health insurance system. PMID- 15333123 TI - No exit: an evaluation of measures of physician attrition. AB - OBJECTIVE: To validate physicians' self-reported intentions to leave clinical practice and the American Medical Association (AMA) Masterfile practice status variable as measures of physician attrition, and to determine predictors of intention to leave, and actual departure from, clinical practice. DATA SOURCES: Survey of specialist physicians in urban California (1998); the AMA Physician Masterfile (2001); and direct ascertainment of physician practice status (2001). STUDY DESIGN: Physicians' intention to leave clinical practice by 2001 (self reported in 1998) was tested as a measure of each physician's actual practice status in 2001 (directly ascertained). Physician practice status according to the 2001 AMA Masterfile was also tested as a measure of physicians' actual practice status in 2001. Multivariate regression was used to predict both physicians' intentions to leave clinical practice and their actual departure. DATA COLLECTION/EXTRACTION METHODS: AMA Masterfile data on 2001 practice status were obtained for 967 of 968 physician respondents to the 1998 survey. Actual practice status for 2001 was directly ascertained for 957. PRINCIPAL FINDINGS: The sensitivity of Masterfile practice status as a measure of actual departure from clinical practice was 9.0 percent, and the positive predictive value was 52.9 percent. Allowing for a two-year reporting lag did not change this substantially. Self-reported intention to leave clinical practice had a sensitivity of 73.3 percent and a positive predictive value of 35.4 percent as a measure of actual departure from practice. The strongest predictor of both intention to leave clinical practice and actual departure from practice was older age. Physician dissatisfaction had a strong association (OR=5.6) with intention to leave clinical practice, but was not associated with actual departure from practice. CONCLUSIONS: Our findings call into question the accuracy of both AMA Masterfile data and physicians' self-reported intentions to leave as measures of physician attrition from clinical practice. Research using these measures should be interpreted with caution. Self-reported intention to leave practice may be more of a proxy for dissatisfaction than an accurate predictor of actual behavior. PMID- 15333124 TI - The alignment and blending of payment incentives within physician organizations. AB - OBJECTIVE: To analyze the blend of retrospective (fee-for-service, productivity based salary) and prospective (capitation, nonproductivity-based salary) methods for compensating individual physicians within medical groups and independent practice associations (IPAs) and the influence of managed care on the compensation blend used by these physician organizations. DATA SOURCES: Of the 1,587 medical groups and IPAs with 20 or more physicians in the United States, 1,104 responded to a one-hour telephone survey, with 627 providing detailed information on physician payment methods. STUDY DESIGN: We calculated the distribution of compensation methods for primary care and specialty physicians, separately, in both medical groups and IPAs. Multivariate regression methods were used to analyze the influence of market and organizational factors on the payment method developed by physician organizations for individual physicians. PRINCIPAL FINDINGS: Within physician organizations, approximately one-quarter of physicians are paid on a purely retrospective (fee-for-service) basis, approximately one quarter are paid on a purely prospective (capitation, nonproductivity-based salary) basis, and approximately one-half on blends of retrospective and prospective methods. Medical groups and IPAs in heavily penetrated managed care markets are significantly less likely to pay their individual physicians based on fee-for-service than are organizations in less heavily penetrated markets. CONCLUSIONS: Physician organizations rely on a wide range of prospective, retrospective, and blended payment methods and seek to align the incentives faced by individual physicians with the market incentives faced by the physician organization. PMID- 15333125 TI - The effect of HMOs on the inpatient utilization of medicare beneficiaries. AB - OBJECTIVE: To determine the effect of joining HMOs (health maintenance organizations) on the inpatient utilization of Medicare beneficiaries. DATA SOURCES: We linked enrollment data on Medicare beneficiaries to patient discharge data from the California Office of Statewide Health Planning and Development (OSHPD) for 1991-1995. DESIGN AND SAMPLE: A quasi-experimental design comparing inpatient utilization before and after switching from fee-for-service (FFS) to Medicare HMOs; with comparison groups of continuous FFS and HMO beneficiaries to adjust for aging and secular trends. The sample consisted of 124,111 Medicare beneficiaries who switched from FFS to HMOs in 1992 and 1993, and random samples of 108,966 continuous FFS beneficiaries and 18,276 continuous HMO enrollees yielding 1,227,105 person-year observations over five years. MAIN OUTCOMES MEASURE: Total inpatient days per thousand per year. PRINCIPAL FINDINGS: When beneficiaries joined a group/staff HMO, their total days per year were 18 percent lower (95 percent confidence interval, 15-22 percent) than if the beneficiaries had remained in FFS. Total days per year were reduced less for beneficiaries joining an IPA (independent practice association) HMO (11 percent; 95 percent confidence interval, 4-19 percent). Medicare group/staff and IPA-model HMO enrollees had roughly 60 percent of the inpatient days per thousand beneficiaries in 1995 as did FFS beneficiaries (976 and 928 versus 1,679 days per thousand, respectively). In the group/staff model HMOs, our analysis suggests that managed care practices accounted for 214 days of this difference, and the remaining 489 days (70 percent) were due to favorable selection. In IPA HMOs, managed care practices appear to account for only 115 days, with 636 days (85 percent) due to selection. CONCLUSIONS: Through the mid-nineties, Medicare HMOs in California were able to reduce inpatient utilization beyond that attributable to the high level of favorable selection, but the reduction varied by type of HMO. PMID- 15333128 TI - Evidence-based health care 2004: AHRQ moves research to translation and implementation. PMID- 15333129 TI - Diurnal difference in CAR mRNA expression. AB - BACKGROUND: The constitutive androstane receptor (CAR, NR1I3) plays a key role in the transcriptional activation of genes that encode xenobiotic/steroid and drug metabolizing enzymes. RESULTS: The expression of CAR mRNA throughout the circadian rhythm is reported for the first time in phase with the clock gene Bmal1 and in antiphase with the clock-controlled gene Rev-erbalpha mRNAs, with a peak at Zeitgeber time (ZT) 20 and a trough at ZT8, and a peak/trough ratio of 2.0. The diurnal difference in CAR mRNA expression might underlie the 1.7-fold difference in the magnitude of the PB-dependent induction of CYP2B1/2 mRNA. CONCLUSION: The circadian oscillation of xenosensor gene CAR mRNA expression is partially responsible for chronopharmacokinetics and chronopharmacology in disease. PMID- 15333130 TI - How do drug users define their progress in harm reduction programs? Qualitative research to develop user-generated outcomes. AB - BACKGROUND: Harm reduction is a relatively new and controversial model for treating drug users, with little formal research on its operation and effectiveness. In order to advance the study of harm reduction programs and our understanding of how drug users define their progress, qualitative research was conducted to develop outcomes of harm reduction programming that are culturally relevant, incremental, (i.e., capable of measuring change), and hierarchical (i.e., capable of showing how clients improve over time). METHODS: The study used nominal group technique (NGT) to develop the outcomes (phase 1) and focus group interviews to help validate the findings (phase 2). Study participants were recruited from a large harm-reduction program in New York City and involved approximately 120 clients in 10 groups in phase 1 and 120 clients in 10 focus groups in phase 2. RESULTS: Outcomes of 10 life areas important to drug users were developed that included between 10 to 15 incremental measures per outcome. The outcomes included ways of 1) making money; 2) getting something good to eat; 3) being housed/homeless; 4) relating to families; 5) getting needed programs/benefits/services; 6) handling health problems; 7) handling negative emotions; 8) handling legal problems; 9) improving oneself; and 10) handling drug use problems. Findings also provided insights into drug users' lives and values, as well as a window into understanding how this population envisions a better quality of life. Results challenged traditional ways of measuring drug users based solely on quantity used and frequency of use. They suggest that more appropriate measures are based on the extent to which drug users organize their lives around drug use and how much drug use is integrated into their lives and negatively impacts other aspects of their lives. CONCLUSIONS: Harm reduction and other programs serving active drug users and other marginalized people should not rely on institutionalized, provider-defined solutions to problems in living faced by their clients. PMID- 15333131 TI - Exploratory disease mapping: kriging the spatial risk function from regional count data. AB - BACKGROUND: There is considerable interest in the literature on disease mapping to interpolate estimates of disease occurrence or risk of disease from a regional database onto a continuous surface. In addition to many interpolation techniques available the geostatistical method of kriging has been used but also criticised. RESULTS: To circumvent these critics one may use kriging along with already smoothed regional estimates, where smoothing is based on empirical Bayes estimates, also known as shrinkage estimates. The empirical Bayes step has the advantage of shrinking the unstable and often extreme estimates to the global or local mean, and also has a stabilising effect on variance by borrowing strength, as well. Negative interpolates are prevented by choice of the appropriate kriging method. The proposed mapping method is applied to the North Carolina SIDS data example as well as to an example data set from veterinary epidemiology. The SIDS data are modelled without spatial trend. And spatial interpolation is based on ordinary kriging. The second example is included to demonstrate the method when the phenomenon under study exhibits a spatial trend and interpolation is based on universal kriging. CONCLUSION: Interpolation of the regional estimates overcomes the areal bias problem and the resulting isopleth maps are easier to read than choropleth maps. The empirical Bayesian estimate for smoothing is related to internal standardization in epidemiology. Therefore, the proposed concept is easily communicable to map users. PMID- 15333132 TI - Real time electrocardiogram QRS detection using combined adaptive threshold. AB - BACKGROUND: QRS and ventricular beat detection is a basic procedure for electrocardiogram (ECG) processing and analysis. Large variety of methods have been proposed and used, featuring high percentages of correct detection. Nevertheless, the problem remains open especially with respect to higher detection accuracy in noisy ECGs METHODS: A real-time detection method is proposed, based on comparison between absolute values of summed differentiated electrocardiograms of one of more ECG leads and adaptive threshold. The threshold combines three parameters: an adaptive slew-rate value, a second value which rises when high-frequency noise occurs, and a third one intended to avoid missing of low amplitude beats. Two algorithms were developed: Algorithm 1 detects at the current beat and Algorithm 2 has an RR interval analysis component in addition. The algorithms are self-adjusting to the thresholds and weighting constants, regardless of resolution and sampling frequency used. They operate with any number L of ECG leads, self-synchronize to QRS or beat slopes and adapt to beat to-beat intervals. RESULTS: The algorithms were tested by an independent expert, thus excluding possible author's influence, using all 48 full-length ECG records of the MIT-BIH arrhythmia database. The results were: sensitivity Se = 99.69 % and specificity Sp = 99.65 % for Algorithm 1 and Se = 99.74 % and Sp = 99.65 % for Algorithm 2. CONCLUSION: The statistical indices are higher than, or comparable to those, cited in the scientific literature. PMID- 15333133 TI - The medical care of patients with primary care home nursing is complex and influenced by non-medical factors: a comprehensive retrospective study from a suburban area in Sweden. AB - BACKGROUND: The reduced number of hospital beds and an ageing population have resulted in growing demands for home nursing. We know very little about the comprehensive care of these patients. The objectives were to identify the care, in addition to primary health care, of patients with primary-care home nursing to give a comprehensive view of their care and to investigate how personal, social and functional factors influence the use of specialised medical care. METHODS: One-third (158) of all patients receiving primary-care home nursing in an area were sampled, and 73 % (116) were included. Their care from October 1995 until October 1996 was investigated by sending questionnaires to district nurses and home-help providers and by collecting retrospective data from primary-care records and official statistics. We used non-parametric statistical methods, i.e. medians and minimum - maximum, chi2, and the Mann-Whitney test, since the data were not normally distributed. Conditional logistic regression was used to study whether personal, social or functional factors influenced the chance (expressed as odds ratio) that study patients had made visits to or had received inpatient care from specialised medical care during the study year. RESULTS: 56 % of the patients had been hospitalised. 73 % had made outpatient visits to specialised medical care. The care took place at 14 different hospitals, and more than 22 specialities were involved, but local care predominated. Almost all patients visited doctors, usually in both primary and specialised medical care. Patients who saw doctors in specialised care had more help from all other categories of care. Patients who received help from their families made more visits to specialised medical care and patients with severe ADL dependence made fewer visits. CONCLUSIONS: The care of patients with primary-care home nursing is complex. Apart from home nursing, all patients also made outpatient visits to doctors, usually in both primary and specialised medical care. Many different caregivers and professions were involved. Reduced functional capacity decreased and help from family members increased the chance of having received outpatient specialised medical care. This raises questions concerning the medical care for patients with both medical and functional problems. PMID- 15333134 TI - Seasonal ovulatory activity exists in tropical Creole female goats and Black Belly ewes subjected to a temperate photoperiod. AB - BACKGROUND: Seasonality of ovulatory activity is observed in European sheep and goat breeds, whereas tropical breeds show almost continuous ovulatory activity. It is not known if these tropical breeds are sensitive or not to temperate photoperiod. This study was therefore designed to determine whether tropical Creole goats and Black-Belly ewes are sensitive to temperate photoperiod. Two groups of adult females in each species, either progeny or directly born from imported embryos, were used and maintained in light-proof rooms under simulated temperate (8 to 16 h of light per day) or tropical (11 - 13 h) photoperiods. Ovulatory activity was determined by blood progesterone assays for more than two years. The experiment lasted 33 months in goats and 25 months in ewes. RESULTS: Marked seasonality of ovulatory activity appeared in the temperate group of Creole female goats. The percentage of female goats experiencing at least one ovulation per month dramatically decreased from May to September for the three years (0%, 27% and 0%, respectively). Tropical female goats demonstrated much less seasonality, as the percentage of goats experiencing at least one ovulation per month never went below 56%. These differences were significant. Both groups of temperate and tropical Black-Belly ewes experienced a marked seasonality in their ovulatory activity, with only a slightly significant difference between groups. The percentage of ewes experiencing at least one ovulation per month dropped dramatically in April and rose again in August (tropical ewes) or September (temperate ewes). The percentage of ewes experiencing at least one ovulation per month never went below 8% and 17% (for tropical and temperate ewes respectively) during the spring and summer months. CONCLUSIONS: An important seasonality in ovulatory activity of tropical Creole goats was observed when females were exposed to a simulated temperate photoperiod. An unexpected finding was that Black-Belly ewes and, to a lesser extent, Creole goats exposed to a simulated tropical photoperiod also showed seasonality in their ovulatory activity. Such results indicate that both species are capable of showing seasonality under the photoperiodic changes of the temperate zone even though they do not originate from these regions. PMID- 15333135 TI - Enhanced diagnostic immunofluorescence using biopsies transported in saline. AB - BACKGROUND: The demonstration of tissue-bound immunoreactants by direct immunofluorescence microscopy (DIF) is a valuable parameter in the diagnosis of various autoimmune and immunecomplex-mediated skin diseases. For preservation of tissue-bound immunoreactants, biopsies are usually fresh-frozen in liquid nitrogen or transported in Michel's fixative. But even optimally preserved tissue specimens are no guarantee for the correct diagnosis by DIF, especially when weak to moderate IgG fluorescence of the epidermal basement membrane zone is involved. In such cases false negative results are easily obtained due to the relatively high dermal "background" fluorescence produced by polyclonal anti-human IgG fluorescein conjugates. METHODS: In the present study we have compared the use of normal saline (0.9% NaCl) with liquid nitrogen and Michel's fixative as transport medium for skin biopsies. From 25 patients with an autoimmune skin disease (pemphigus, pemphigoid, lupus erythematosus and vasculitis) four matched skin biopsies were obtained and transported in either saline for 24 and 48 hours, liquid nitrogen, or Michel's fixative for 48 hours. RESULTS: Direct IF microscopy showed significant reduction of background fluorescence (p < 0.01) and relatively enhanced desired specific (IgG, IgA) staining in biopsies transported in saline. A conclusive or tentative IF diagnosis was reached in 92% after 24 h saline, 83% after 48 h saline, 68% after freezing in liquid nitrogen, and 62% after 48 h Michel's medium (n = 25). CONCLUSIONS: We conclude that transporting biopsies without freezing in normal saline for 24 hours is an adequate and attractive method for routine IF diagnosis in autoimmune and immune complex-mediated dermatoses. The superior results with saline incubation are explained by washing away of IgG background in dermis and epidermis. PMID- 15333136 TI - Chronic arthritis in children and adolescents in two Indian health service user populations. AB - BACKGROUND: High prevalence rates for rheumatoid arthritis, spondyloarthopathies, and systemic lupus erythematosus have been described in American Indian and Alaskan Native adults. The impact of these diseases on American Indian children has not been investigated. METHODS: We used International Classification of Diseases-9 (ICD-9) codes to search two Indian Health Service (IHS) patient registration databases over the years 1998-2000, searching for individuals 19 years of age or younger with specific ICD-9-specified diagnoses. Crude estimates for disease prevalence were made based on the number of individuals identified with these diagnoses within the database. RESULTS: Rheumatoid arthritis (RA) / juvenile rheumatoid arthritis (JRA) was the most frequent diagnosis given. The prevalence rate for JRA in the Oklahoma City Area was estimated as 53 per 100,000 individuals at risk, while in the Billings Area, the estimated prevalence was nearly twice that, at 115 per 100,000. These rates are considerably higher than those reported in the most recent European studies. CONCLUSION: Chronic arthritis in childhood represents an important, though unrecognized, chronic health challenge within the American Indian population living in the United States. PMID- 15333137 TI - Valgus and varus deformity after wide-local excision, brachytherapy and external beam irradiation in two children with lower extremity synovial cell sarcoma: case report. AB - BACKGROUND: Limb-salvage is a primary objective in the management of extremity soft-tissue sarcoma in adults and children. Wide-local excision combined with radiation therapy is effective in achieving local tumor control with acceptable morbidity and good functional outcomes for most patients. CASE PRESENTATION: Two cases of deformity after wide-local excision, brachytherapy and external beam irradiation for lower-extremity synovial cell sarcoma are presented and discussed to highlight contributing factors, time course of radiation effects and orthopedic management. In an effort to spare normal tissues from the long-term effects of radiation therapy, more focal irradiation techniques have been applied to patients with musculoskeletal tumors including brachytherapy and conformal radiation therapy. As illustrated in this report, the use of these techniques results in the asymmetric irradiation of growth plates and contributes to the development of valgus or varus deformity and leg-length discrepancies. CONCLUSIONS: Despite good functional outcomes, progressive deformity in both patients required epiphysiodesis more than 3 years after initial management. There is a dearth of information related to the effects of radiation therapy on the musculoskeletal system in children. Because limb-sparing approaches are to be highlighted in the next generation of cooperative group protocols for children with musculoskeletal tumors, documentation of the effects of surgery and radiation therapy will lead to improved decision making in the selection of the best treatment approach and in the follow-up of these patients. PMID- 15333138 TI - Malaria and anemia prevention in pregnant women of rural Burkina Faso. AB - BACKGROUND: Pregnant women are a major risk group for malaria in endemic areas. Only little information exists on the compliance of pregnant women with malaria and anaemia preventive drug regimens in the rural areas of sub-Saharan Africa (SSA). In this study, we collected information on malaria and anaemia prevention behaviour in pregnant women of rural Burkina Faso. METHODS: Cross-sectional qualitative and quantitative survey among 225 women of eight villages in rural northwestern Burkina Faso. Four of the villages had a health centre offering antenatal care (ANC) services while the other four were more than five kilometers away from a health centre. RESULTS: Overall ANC coverage (at least one visit) was 71% (95% in health centre villages vs 50% in remote villages). Malaria and anaemia were considered as the biggest problems during pregnancy in this community. ANC using women were quite satisfied with the quality of services, and compliance with malaria and anaemia prevention regimens (chloroquine and iron/folic acid) was high in this population. Knowledge on the benefit of bed nets and good nutrition was less prominent. Distance, lack of money and ignorance were the main reasons for women to not attend ANC services. CONCLUSIONS: There is an urgent need to improve access of rural SSA women to ANC services, either through increasing the number of rural health centres or establishing functioning outreach services. Moreover, alternative malaria and anaemia prevention programmes such as intermittent preventive treatment with effective antimalarials and the distribution of insecticide-treated bed nets need to become implemented on a large scale. PMID- 15333139 TI - Spontaneous activity of rat pretectal nuclear complex neurons in vitro. AB - BACKGROUND: Neurons in the mammalian pretectum are involved in the control of various visual and oculomotor tasks. Because functionally independent pretectal cell populations show a wide variation of response types to visual stimulation in vivo, they may also differ in their intrinsic properties when recorded in vitro. We therefore performed whole-cell patch clamp recordings from neurons in the caudal third of the pretectal nuclear complex in frontal brain slices obtained from 3 to 6 week old hooded rats and tried to classify pretectal neurons electrophysiologically. RESULTS: Pretectal neurons showed various response types to intracellular depolarizations, including bursting and regular firing behavior. One population of pretectal nuclear complex neurons could be particularly distinguished from others because they displayed spontaneous activity in vitro. These cells had more positive resting potentials and higher input resistances than cells that were not spontaneously active. The maintained firing of spontaneously active pretectal cells was characterized by only small variances in interspike intervals and thus showed a regular temporal patterning. The firing rate was directly correlated to the membrane potential. Removing excitatory inputs by blockade of AMPA and/or NMDA receptors did not change the spontaneous activity. Simultaneous blockade of excitatory and inhibitory synaptic input by a substitution of extracellular calcium with cobalt neither changed the firing rate nor its temporal patterning. Each action potential was preceeded by a depolarizing inward current which was insensitive to calcium removal but which disappeared in the presence of tetrodotoxin. CONCLUSIONS: Our results indicate that a specific subpopulation of pretectal neurons is capable of generating maintained activity in the absence of any external synaptic input. This maintained activity depends on a sodium conductance and is independent from calcium currents. PMID- 15333140 TI - Sequence periodicity of Escherichia coli is concentrated in intergenic regions. AB - BACKGROUND: Sequence periodicity with a period close to the DNA helical repeat is a very basic genomic property. This genomic feature was demonstrated for many prokaryotic genomes. The Escherichia coli sequences display the period close to 11 base pairs. RESULTS: Here we demonstrate that practically only ApA/TpT dinucleotides contribute to overall dinucleotide periodicity in Escherichia coli. The noncoding sequences reveal this periodicity much more prominently compared to protein-coding sequences. The sequence periodicity of ApC/GpT, ApT and GpC dinucleotides along the Escherichia coli K-12 is found to be located as well mainly within the intergenic regions. CONCLUSIONS: The observed concentration of the dinucleotide sequence periodicity in the intergenic regions of E. coli suggests that the periodicity is a typical property of prokaryotic intergenic regions. We suppose that this preferential distribution of dinucleotide periodicity serves many biological functions; first of all, the regulation of transcription. PMID- 15333141 TI - POSA: perl objects for DNA sequencing data analysis. AB - BACKGROUND: Capillary DNA sequencing machines allow the generation of vast amounts of data with little hands-on time. With this expansion of data generation, there is a growing need for automated data processing. Most available software solutions, however, still require user intervention or provide modules that need advanced informatics skills to allow implementation in pipelines. RESULTS: Here we present POSA, a pair of new perl objects that describe DNA sequence traces and Phrap contig assemblies in detail. Methods included in POSA include basecalling with quality scores (by Phred), contig assembly (by Phrap), generation of primer3 input and automated SNP annotation (by PolyPhred). Although easily implemented by users with only limited programming experience, these objects considerabily reduce hands-on analysis time compared to using the Staden package for extracting sequence information from raw sequencing files and for SNP discovery. CONCLUSIONS: The POSA objects allow a flexible and easy design, implementation and usage of perl-based pipelines to handle and analyze DNA sequencing data, while requiring only minor programming skills. PMID- 15333142 TI - Molecular clock in neutral protein evolution. AB - BACKGROUND: A frequent observation in molecular evolution is that amino-acid substitution rates show an index of dispersion (that is, ratio of variance to mean) substantially larger than one. This observation has been termed the overdispersed molecular clock. On the basis of in silico protein-evolution experiments, Bastolla and coworkers recently proposed an explanation for this observation: Proteins drift in neutral space, and can temporarily get trapped in regions of substantially reduced neutrality. In these regions, substitution rates are suppressed, which results in an overall substitution process that is not Poissonian. However, the simulation method of Bastolla et al. is representative only for cases in which the product of mutation rate micro and population size Ne is small. How the substitution process behaves when micro Ne is large is not known. RESULTS: Here, I study the behavior of the molecular clock in in silico protein evolution as a function of mutation rate and population size. I find that the index of dispersion decays with increasing micro Ne, and approaches 1 for large micro Ne. This observation can be explained with the selective pressure for mutational robustness, which is effective when micro Ne is large. This pressure keeps the population out of low-neutrality traps, and thus steadies the ticking of the molecular clock. CONCLUSIONS: The molecular clock in neutral protein evolution can fall into two distinct regimes, a strongly overdispersed one for small micro Ne, and a mostly Poissonian one for large micro Ne. The former is relevant for the majority of organisms in the plant and animal kingdom, and the latter may be relevant for RNA viruses. PMID- 15333143 TI - Different patterns of evolution for duplicated DNA repair genes in bacteria of the Xanthomonadales group. AB - BACKGROUND: DNA repair genes encode proteins that protect organisms against genetic damage generated by environmental agents and by-products of cell metabolism. The importance of these genes in life maintenance is supported by their high conservation, and the presence of duplications of such genes may be easily traced, especially in prokaryotic genomes. RESULTS: The genome sequences of two Xanthomonas species were used as the basis for phylogenetic analyses of genes related to DNA repair that were found duplicated. Although 16S rRNA phylogenetic analyses confirm their classification at the basis of the gamma proteobacteria subdivision, differences were found in the origin of the various genes investigated. Except for lexA, detected as a recent duplication, most of the genes in more than one copy are represented by two highly divergent orthologs. Basically, one of such duplications is frequently positioned close to other gamma proteobacteria, but the second is often positioned close to unrelated bacteria. These orthologs may have occurred from old duplication events, followed by extensive gene loss, or were originated from lateral gene transfer (LGT), as is the case of the uvrD homolog. CONCLUSIONS: Duplications of DNA repair related genes may result in redundancy and also improve the organisms' responses to environmental challenges. Most of such duplications, in Xanthomonas, seem to have arisen from old events and possibly enlarge both functional and evolutionary genome potentiality. PMID- 15333144 TI - Mutations in the IGF-II pathway that confer resistance to lytic reovirus infection. AB - BACKGROUND: Viruses are obligate intracellular parasites and rely upon the host cell for different steps in their life cycles. The characterization of cellular genes required for virus infection and/or cell killing will be essential for understanding viral life cycles, and may provide cellular targets for new antiviral therapies. RESULTS: A gene entrapment approach was used to identify candidate cellular genes that affect reovirus infection or virus induced cell lysis. Four of the 111 genes disrupted in clones selected for resistance to infection by reovirus type 1 involved the insulin growth factor-2 (IGF-II) pathway, including: the mannose-6-phosphate/IGF2 receptor (Igf2r), a protease associated with insulin growth factor binding protein 5 (Prss11), and the CTCF transcriptional regulator (Ctcf). The disruption of Ctcf, which encodes a repressor of Igf2, was associated with enhanced Igf2 gene expression. Plasmids expressing either the IGF-II pro-hormone or IGF-II without the carboxy terminal extension (E)-peptide sequence independently conferred high levels of cellular resistance to reovirus infection. Forced IGF-II expression results in a block in virus disassembly. In addition, Ctcf disruption and forced Igf2 expression both enabled cells to proliferate in soft agar, a phenotype associated with malignant growth in vivo. CONCLUSION: These results indicate that IGF-II, and by inference other components of the IGF-II signalling pathway, can confer resistance to lytic reovirus infection. This report represents the first use of gene entrapment to identify host factors affecting virus infection. Concomitant transformation observed in some virus resistant cells illustrates a potential mechanism of carcinogenesis associated with chronic virus infection. PMID- 15333145 TI - XHM: a system for detection of potential cross hybridizations in DNA microarrays. AB - BACKGROUND: Microarrays have emerged as the preferred platform for high throughput gene expression analysis. Cross-hybridization among genes with high sequence similarities can be a source of error reducing the reliability of DNA microarray results. RESULTS: We have developed a tool called XHM (cross hybridization on microarrays) for assessment of the reliability of hybridization signals by detecting potential cross-hybridizations on DNA microarrays. This is done by comparing the sequences of the probes against an extensive database representing the transcriptome of the organism in question. XHM is available online at http://www.bioinfo.no/tools/xhm/. CONCLUSIONS: Using XHM with its user adjustable parameters will enable scientists to check their lists of differentially expressed genes from microarray experiments for potential cross hybridizations. This provides information that may be useful in the validation of the microarray results. PMID- 15333146 TI - Applying Support Vector Machines for Gene Ontology based gene function prediction. AB - BACKGROUND: The current progress in sequencing projects calls for rapid, reliable and accurate function assignments of gene products. A variety of methods has been designed to annotate sequences on a large scale. However, these methods can either only be applied for specific subsets, or their results are not formalised, or they do not provide precise confidence estimates for their predictions. RESULTS: We have developed a large-scale annotation system that tackles all of these shortcomings. In our approach, annotation was provided through Gene Ontology terms by applying multiple Support Vector Machines (SVM) for the classification of correct and false predictions. The general performance of the system was benchmarked with a large dataset. An organism-wise cross-validation was performed to define confidence estimates, resulting in an average precision of 80% for 74% of all test sequences. The validation results show that the prediction performance was organism-independent and could reproduce the annotation of other automated systems as well as high-quality manual annotations. We applied our trained classification system to Xenopus laevis sequences, yielding functional annotation for more than half of the known expressed genome. Compared to the currently available annotation, we provided more than twice the number of contigs with good quality annotation, and additionally we assigned a confidence value to each predicted GO term. CONCLUSIONS: We present a complete automated annotation system that overcomes many of the usual problems by applying a controlled vocabulary of Gene Ontology and an established classification method on large and well-described sequence data sets. In a case study, the function for Xenopus laevis contig sequences was predicted and the results are publicly available at ftp://genome.dkfz heidelberg.de/pub/agd/gene_association.agd_Xenopus. PMID- 15333147 TI - Vernon Young and the development of current knowledge in protein and amino acid nutrition. PMID- 15333148 TI - Religion, diet and research. PMID- 15333150 TI - Prospective evaluation of lactose malabsorption by lactose hydrogen breath test in individuals infected with Entamoeba histolytica and passing cysts. AB - The aim of the present prospective study was to detect lactose malabsorption in subjects in northern India infected with Entamoeba histolytica and passing cysts. The study group included forty-one patients with E. histolytica cysts in at least one of three consecutive faecal samples. Lactose malabsorption was detected by a lactose H2 breath test. The results were compared with those of forty controls subjects. Thirty-two of forty-one (78.0 %) subjects passing E. histolytica cysts had lactose malabsorption compared with seventeen of forty (42.5 %) control subjects (P<0.01). In conclusion, the present study shows that lactose malabsorption is significantly more common in individuals infected with E. histolytica and passing cysts compared with control subjects. PMID- 15333149 TI - Plasma malondialdehyde is induced by smoking: a study with balanced antioxidant profiles. AB - It has been reported that smokers have higher plasma malondialdehyde concentrations compared with non-smokers. However, smokers have also consistently been shown to have a lower intake of fruits and vegetables as well as lower plasma antioxidant concentrations. Since both the latter issues may well influence the malondialdehyde concentration, we wanted to investigate if the observed difference between smokers and non-smokers was a result of differences in antioxidant status or if a more direct effect of smoking could also be isolated. In the present study, the plasma malondialdehyde and antioxidant profiles of a cohort of smokers (n 48) and non-smokers (n 32) were compared. While there was no significant difference in the major plasma antioxidants measured, i.e. ascorbic acid, alpha- and gamma-tocopherol and uric acid, we found a significant effect of smoking on plasma malondialdehyde (P=0.0003). Consequently, the present study suggests that lipid peroxidation as measured by plasma malondialdehyde is induced by smoking per se. While poor antioxidant status presumably also affects lipid peroxidation, it is only partly responsible for the increased level found in smokers in general. PMID- 15333151 TI - Effects of beta-carotene on cell viability and antioxidant status of hepatocytes from chronically ethanol-fed rats. AB - The purpose of the present study was to evaluate the effects of beta-carotene on the cell viability and antioxidant status of hepatocytes from chronically ethanol fed rats. Rats in the ethanol group were given an ethanol-containing liquid diet that provided 36 % of total energy as ethanol, while rats in the control group were fed an isoenergetic diet without ethanol. After 4 weeks, hepatocytes were taken out and cultured for 24 h. Hepatocytes from the rats in the control and ethanol groups were cultured in medium without (HC, HE) or with beta-carotene (HC+B, HE+B). The results showed that lactate dehydrogenase leakage was significantly increased in the HE compared with that in the HC group. However, lactate dehydrogenase leakage of the HE+B group was similar to that of the HC group. When compared with the HC group, activities of glutathione peroxidase and catalase in the HE group were significantly decreased by 54 and 31 %, respectively. Catalase activity in the HE+B group was significantly increased by 61 % compared with that in the HE group. However, activities of glutathione reductase and superoxide dismutase showed no difference among the groups. The level of glutathione in the HC+B and HE+B groups was significantly increased to 155 and 143 % compared with those in the HC and HE groups, respectively. The concentration of lipid peroxides showed no difference among the groups. The present results demonstrate that beta-carotene improved the cell viability of hepatocytes, and increased catalase activities and glutathione levels in hepatocytes from chronically ethanol-fed rats. PMID- 15333152 TI - Endogenous amino acid flow in the avian ileum: quantification using three techniques. AB - The aim of the present study was to compare the protein-free diet, guanidinated casein (GuC) and enzyme hydrolysed casein (EHC) methods for the quantification of endogenous amino acid (AA) flow in the avian ileum. Growing broiler chickens (5 weeks old) were used. All three assay diets were based on dextrose, and in the GuC and EHC diets GuC or EHC were the sole source of N. Endogenous AA flows determined with the use of protein-free diet were considerably lower (P<0.05) than those determined by the GuC and EHC methods. The total endogenous AA flows determined by the GuC and EHC methods were almost 3-fold greater (P<0.05) than those determined by the protein-free diet. The endogenous AA values obtained from GuC and EHC methods were similar (P>0.05), except for the flow of arginine, which was lower (P<0.05) in the EHC method. Glutamic acid, aspartic acid, threonine and glycine were the predominant endogenous AA present in digesta from the distal ileum. The contents of methionine, histidine and cystine were lower compared with other AA. The method of determination had no effect on the AA composition of endogenous protein, except for threonine, glutamic acid, lysine, arginine and cystine. The concentrations of threonine and arginine were lower (P<0.05) and that of lysine was higher (P<0.05) with the EHC method compared with the other two methods. The concentration of glutamic acid was greater (P<0.05) and that of cystine was lower (P<0.05) in the EHC and GuC methods compared with the protein free diet method. The results showed that the ileal endogenous flows of N and AA are markedly enhanced by the presence of protein and peptides, above those determined following feeding of a protein-free diet. It is concluded that the use of EHC and GuC methods enables the measurement of ileal endogenous losses in chickens under normal physiological conditions. PMID- 15333153 TI - Urinary excretion of purine derivatives as an index of microbial protein synthesis in the camel (Camelus dromedarius). AB - Five experiments were carried out to extend knowledge of purine metabolism in the camel (Camelus dromedarius) and to establish a model to enable microbial protein outflow from the forestomachs to be estimated from the urinary excretion of purine derivatives (PD; i.e. xanthine, hypoxanthine, uric acid, allantoin). In experiment 1, four camels were fasted for five consecutive days to enable endogenous PD excretion in urine to be determined. Total PD excretion decreased during the fasting period to 267 (SE 41.5) micromol/kg body weight (W)0.75 per d. Allantoin and xanthine + hypoxanthine were consistently 86 and 6.1 % of total urinary PD during this period but uric acid increased from 3.6 % to 7.4 %. Xanthine oxidase activity in tissues (experiment 2) was (micromol/min per g fresh tissue) 0.038 in liver and 0.005 in gut mucosa but was not detected in plasma. In experiment 3, the duodenal supply of yeast containing exogenous purines produced a linear increase in urinary PD excretion rate with the slope indicating that 0.63 was excreted in urine. After taking account of endogenous PD excretion, the relationship can be used to predict purine outflow from the rumen. From the latter prediction, and also the purine:protein ratio in bacteria determined in experiment 5, we predicted the net microbial outflow from the rumen. In experiment 4, with increasing food intake, the rate of PD excretion in the urine increased linearly by about 11.1 mmol PD/kg digestible organic matter intake (DOMI), equivalent to 95 g microbial protein/kg DOMI. PMID- 15333154 TI - The ability of laying hens to regulate phosphorus intake when offered two feeds containing different levels of phosphorus. AB - The ability of laying hens to adjust their intake of available P (AP) was investigated with a maize-soyabean diet fed to forty-eight individually caged birds in a 2 x 4 factorial experiment. From 19 to 25 weeks of age (phase 1) twenty-four birds were fed a normal-P (NP) diet (2.2 g AP/kg DM) and twenty-four were fed a low-P (LP) diet (1.1 g AP/kg). LP eggs were lighter (51 v. 54 (SEM 1.0) g; P<0.05), providing evidence that the LP diet was deficient in AP. From 25 to 28 weeks of age six hens from each phase 1 treatment were fed either the NP or LP diet alone or a choice of the LP and NP feeds or a choice of the LP feed and a phytase-supplemented (PP) feed (LP diet with 400 microbial phytase units/kg). With a choice of the NP and LP feeds, the hens fed the LP diet in phase 1 ate a smaller proportion of the LP feed (34 (SEM 12.0) %) than the hens fed the NP diet in phase 1 (72 (SEM 12.0) %; P<0.05), showing that P deficiency influenced subsequent selection for AP, i.e. an appetite for P was demonstrated. In those birds offered the LP and PP feeds, the presence of phytase in one of the two feeds significantly alleviated the effect of P deficiency on egg and body weights. The proportion of the LP diet chosen was not significantly affected by phase 1 treatment; it was not necessary for the hens to eat more than 50 % of PP feed. PMID- 15333155 TI - Studies on the bioavailability of the provitamin A carotenoid, beta-carotene, using human exfoliated colonic epithelial cells. AB - The possibility of using exfoliated colonic epithelial cells for assessing the bioavailability of beta-carotene was examined. Analysis of exfoliated colonic epithelial cells showed the presence of beta-carotene and vitamin A. The beta carotene content was significantly lower in cells from stool samples of subjects on a beta-carotene-poor diet than those receiving a single dose of a beta carotene supplement. Colonic epithelial cells isolated from stool samples collected daily during a wash-out period while the subjects were on a beta carotene-poor diet showed a steady decrease in beta-carotene content, reaching the lowest value on day 7. Kinetic analysis showed that a single dose of a beta carotene supplement in the form of spirulina (Spirulina platensis) or agathi (Sesbania grandiflora) after the wash-out period caused an increase in the beta carotene content after a lag period of 5-7 d, but the vitamin A levels during these periods were not significantly affected. Analysis of plasma beta-carotene concentration also showed similar changes, which correlated with those of exfoliated colonic cells. A relationship between the beta-carotene content of the diet and that of the colonic epithelial cells suggests that analysis of the beta carotene content in exfoliated human colonic epithelial cells is a useful non invasive method to assess the bioavailability of provitamin A beta-carotene. PMID- 15333156 TI - Reduction of allergic airway eosinophilia by dietary raffinose in Brown Norway rats. AB - Oral administration of raffinose, a naturally occurring indigestible oligosaccharide, has reportedly ameliorated atopic dermatitis in human subjects although the mechanism is unknown. The present study investigated the effect of dietary raffinose on allergen-induced airway eosinophilia in ovalbumin-sensitised Brown Norway rats as an atopic disease model. Brown Norway rats were immunised by subcutaneous injection with ovalbumin on day 0 and fed either a control diet or the diet supplemented with raffinose (50 g/kg diet). The rats were exposed to aerosolised ovalbumin on day 20, and broncho-alveolar lavage fluid was obtained on the next day. The number of eosinophils in the fluid was significantly lower in the rats fed the raffinose diet than in those fed the control diet. Dietary raffinose significantly reduced IL-4 and IL-5 mRNA levels in lung tissue and tended to lower ovalbumin-specific Ig E levels. Suppression of eosinophilia by dietary raffinose was still observed in caecectomised and neomycin-administered rats, suggesting little contribution by the colonic bacteria to the effect of raffinose. Intraperitoneal administration of raffinose also suppressed eosinophilia. Significant concentrations of raffinose were detected in portal venous and abdominal arterial plasma after the intragastric administration of raffinose. Overall, the findings suggest that dietary raffinose ameliorates allergic airway eosinophilia at least partly via post-absorptive mechanisms in Brown Norway rats. PMID- 15333157 TI - Thermally oxidised sunflower-seed oil increases liver and serum peroxidation and modifies lipoprotein composition in rats. AB - Peroxidation of LDL and other lipoproteins is thought to play a central role in atherogenesis. Dietary thermally oxidised oils may increase atherogenic risk in consumers by increasing their oxidative status. The present paper compares the effects of two diets containing unused sunflower-seed oil (US) or sunflower-seed oil repeatedly used in frying (FS) (both 15 g/100 g diet) on weight gain, food efficiency ratio, serum lipid levels and lipoprotein composition, and the content of thiobarbituric acid-reactive substances (TBARS) in the liver, serum, and lipoproteins in growing Wistar rats. After sixty potato fryings the FS contained 27.7 g polar material/100 g oil and 16.6 g oligomers/100 g oil. The FS-fed rats had a significantly lower weight gain and food efficiency ratio. Liver-TBARS increased due to the consumption of the highly altered oil and showed a significant linear relationship (all r > 0.68; P < 0.002) with the ingestion of thermally oxidised compounds. Serum-, VLDL-, LDL- and HDL-TBARS were significantly higher in the FS-fed rats (all P < 0.001). Concentrations of serum total and non-esterified cholesterol and phospholipids were significantly higher in the FS-fed rats (P < 0.05, P < 0.05, and P < 0.001, respectively). Serum triacylglycerol concentrations did not vary between the two dietary groups. Total and esterified cholesterol and phospholipid levels increased significantly in the HDL fraction (P < 0.05, P < 0.05, and P < 0.001, respectively) of the FS-fed rats. HDL-cholesterol and HDL-phospholipids were significantly correlated with liver-TBARS (r > 0.747; P < 0.0001), VLDL-TBARS (r > 0.642; P < 0.003), LDL-TBARS (r > 0.475; P < 0.04), and HDL-TBARS (r > 0.787; P < 0.0001). The data suggest that the rat increases HDL as a protecting mechanism against the peroxidative stress induced by the consumption of a diet containing the thermally oxidised oil. PMID- 15333158 TI - Effects of peroxidation products in thermoxidised dietary oil in female rats during rearing, pregnancy and lactation on their reproductive performance and the antioxidative status of their offspring. AB - The present study was performed to investigate whether lipid peroxidation products in thermoxidised dietary oil fed during rearing, pregnancy and lactation influences the reproductive performance of female rats and the antioxidant status of their offspring. Twenty-four female rats were divided into two groups at 4 weeks of age. They were fed diets containing fresh or oxidised oil (the latter prepared by heating at a temperature of 50 degrees C for 16 d) for 14 weeks. At the age of 12 weeks female rats were mated. The number of total pups and pups born alive was not different between both groups. However, individual pups and litters of dams fed oxidised oil were lighter at birth and gained less weight during the suckling period than those of dams fed fresh oil (P < 0.05). Pups of dams fed oxidised oil contained less protein and more fat in their carcasses than those of dams fed fresh oil (P < 0.05). The milk of dams fed oxidised oil had a lower concentration of triacylglycerols and a lower energy content than that of dams fed the fresh oil (P < 0.05). The pups of dams fed oxidised oil had higher concentrations of lipid peroxidation products in the liver at birth and day 19 of lactation than those of dams fed fresh oil (P < 0.05). In conclusion, the present study shows that feeding oxidised oil with a high concentration of lipid peroxidation products to female rats during rearing, pregnancy and lactation influences the development and antioxidant status of fetus and suckling pups. PMID- 15333159 TI - Greek Orthodox fasting rituals: a hidden characteristic of the Mediterranean diet of Crete. AB - The longevity and excellent health status of the population of Crete has been attributed to its lifestyle and dietary habits. The impact of Greek Orthodox Christian Church fasting on these dietary habits has never been studied. One hundred and twenty Greek Orthodox Christians living in Crete participated in a 1 year prospective study. One half of the subjects, who fasted regularly (fasters), and sixty non-faster controls were followed longitudinally for the three main fasting periods over 1 year; Christmas (40 d), Lent (48 d) and the Assumption (15 d). Pre- and end-holy days measurements were performed in each fasting period including: 24 h dietary recall, blood collection and anthropometric measurements. Based on the 24 h recall, fasters as compared with controls had lower intakes of end-holy days dietary cholesterol, total fat, saturated fatty acids, trans-fatty acids and protein (P < 0.001). Fasters presented a decrease of 753 kJ (180 kcal) in end-holy days energy intake (P < 0.05) compared with an increase of 573 kJ (137 kcal) in the controls (P < 0.05). Fasters had a decrease in end-holy days Ca intake (P < 0.001) and an increase in end-holy days total dietary fibre (P < 0.001) and folate (P < 0.05), attributed to their higher consumption of fruit and vegetables in end-holy periods (P < 0.001). There were no differences for other vitamins or minerals between pre- and end-holy periods in both groups except for vitamin B2. The Orthodox Christian dietary regulations are an important component of the Mediterranean diet of Crete characterised by low levels of dietary saturated fatty acids, high levels of fibre and folate, and a high consumption of fruit, vegetables and legumes. PMID- 15333160 TI - Effect of a nutritional intervention promoting the Mediterranean food pattern on electrophoretic characteristics of low-density lipoprotein particles in healthy women from the Quebec City metropolitan area. AB - The objective of the present study was to evaluate the effect of a nutritional intervention promoting the Mediterranean food pattern in free-living conditions on LDL electrophoretic characteristics in a group of seventy-one healthy women, aged between 30 and 65 years. The 12-week nutritional intervention consisted of two courses on nutrition and seven individual sessions with a dietitian. The first course provided information on the Mediterranean food pattern and the second was a cooking lesson. LDL peak particle diameter (LDL-PPD) and cholesterol levels in small (LDL-cholesterol<255 A) and large LDL fractions (LDL cholesterol>260 A) were obtained by 2-16 % polyacrylamide gel electrophoresis of whole plasma. The sample was divided on the basis of baseline LDL-PPD using tertiles of the distribution (258.4 A and 260.0 A). Among the total sample of women, no significant change in LDL-PPD was observed in response to the nutritional intervention. However, subjects who at baseline were in the first tertile of the LDL-PPD distribution (<258.4 A) showed a significant increase in LDL-PPD and in the proportion of LDL %>260 A in response to the 12-week nutritional intervention (P<0.05). In contrast, LDL-PPD decreased significantly (P=0.007) among women with large LDL particles at baseline (LDL-PPD >260 A) while the proportion of LDL %<255 A and of LDL %>260 A remained unchanged. To conclude, changes in the food pattern, in response to a nutritional intervention promoting the Mediterranean food pattern, were accompanied by beneficial modifications in LDL electrophoretic characteristics in women who were characterised at baseline by smaller LDL particles. PMID- 15333161 TI - Distribution of alpha-tocopherol stereoisomers in liver and thigh of chickens. AB - The effect of supplementation with different levels of all-rac-alpha-tocopheryl acetate and the inclusion of different dietary contents of PUFA on the deposition of alpha-tocopherol stereoisomers in liver and thigh of chickens was evaluated. Ninety-six 1-d-old Ross female broiler chickens were randomly distributed into eight experimental treatments (three replicates each) resulting from four levels of alpha-tocopheryl acetate without supplementation and supplemented with 100, 200 and 400 mg alpha-tocopheryl acetate/kg and two levels of dietary PUFA (15 and 61 g/kg). The feeds supplemented with alpha-tocopheryl acetate contained a similar proportion of each stereoisomer. The diets without alpha-tocopheryl acetate had the following alpha-tocopherol stereoisomers (%): RRR 35.1, RRS 24.5, RSR 25.3, RSS 13.9 and total 2S forms 1.3. Consumption of different levels of alpha-tocopheryl acetate did not lead to statistical differences in alpha tocopherol stereoisomer proportion in the liver and thigh. In general, the stereoisomer profiles in the tissues studied were similar, responding to the stereoisomer profile of the diet. Both tissues preferentially accumulated 2R stereoisomer (69-100 %). However, when alpha-tocopheryl acetate was used the discrimination was not specific for the RRR alpha-tocopherol form. Furthermore, the 2R:2S ratio had a tendency to increase as the polyunsaturation level of the diet increased. PMID- 15333162 TI - Effects of dietary sunflower seeds on rumen protozoa and growth of lambs. AB - Linoleic acid-rich sunflower-seed supplements (SSS) were used in two experiments (experiment 1, high-concentrate diets; experiment 2, high-forage diets) to study effects on rumen protozoa and the growth of lambs. Both experiments consisted of four treatments, two with a low-protein diet (120 g/kg) and two with a high protein diet (160 g/kg). For both diets, one treatment was without (control) and one with the SSS (140 g/kg dietary DM). The lambs were fed ad libitum for 70 and 140 d in experiments 1 and 2, respectively. Thereafter, the digestibility of organic matter (OM), acid-detergent fibre and neutral-detergent fibre were determined for each diet with four lambs, and then all lambs were slaughtered and rumen fluid samples were collected and analysed. The results showed substantial decreases (P < 0.001) or total elimination of protozoa in the rumen fluid of the SSS-receiving lambs. In the first experiment the SSS also decreased (P < 0.05) feed intake, but an increase in average daily gain (P < 0.06) resulted in an improved (P < 0.05) feed:gain ratio. Also, the SSS increased (P < 0.05) the digestibility of fibre. In the second experiment the SSS decreased (P < 0.05) the OM digestibility, feed intake and growth of lambs. It was concluded that the use of sunflower-seed supplementation in high-concentrate diets of ruminants reduces rumen fauna, resulting in savings on dietary protein supplements and an increased digestion of feed. PMID- 15333163 TI - Fruit and vegetable consumption is inversely associated with blood pressure in a Mediterranean population with a high vegetable-fat intake: the Seguimiento Universidad de Navarra (SUN) Study. AB - There is evidence that a diet rich in fruit and vegetables reduces blood pressure (BP). Characteristically, the Mediterranean diet is rich in plant-derived foods and also in fat, but studies conducted in Mediterranean countries to relate diet to BP are scarce. We studied the association between fruit and vegetable consumption and BP in a cross-sectional analysis of 4393 participants in the Seguimiento Universidad de Navarra (SUN) Study, an ongoing dynamic cohort study in Spain. Diet was measured using a food-frequency questionnaire previously validated in Spain. Fat represented more than 37 % total energy intake. Subjects were considered to have undiagnosed hypertension if they reported systolic BP > or = 140 mmHg or diastolic BP > or = 90 mmHg, and not a medical diagnosis of hypertension. The adjusted prevalence odds ratio of undiagnosed hypertension (upper v. lowest quintile) was 0.58 (95 % CI 0.36, 0.91; P for trend 0.01) for vegetable consumption and 0.68 (95 % CI 0.43, 1.09; P for trend 0.10) for fruit consumption. Comparing those in the highest quintile of both fruit and vegetable consumption with those in the lowest quintile of both food groups, the prevalence odds ratio was 0.23 (95 % CI 0.10, 0.55; P = 0.001), after adjusting for risk factors for hypertension and other dietary exposures. In a Mediterranean population with an elevated fat consumption, a high fruit and vegetable intake is inversely associated with BP levels. PMID- 15333164 TI - Changes over 20 years in macronutrient intake and body mass index in 11- to 12 year-old adolescents living in Northumberland. AB - Monitoring adolescent diets over time enables the assessment of the effectiveness of public health messages which are particularly important in vulnerable groups such as adolescents. In 2000, 424 children aged 11-12 years old completed two 3 d estimated dietary records. On the fourth day one nutritionist interviewed each child to clarify the information in the diary and foods were quantified with the aid of food models. Nutrient intake was calculated using computerised food tables. These children attended the same seven schools in the same Northumberland area as the 11- to 12-year-old children who recorded their diet using the same method in 1980 (n 405) and 1990 (n 379), respectively. Height and weight, and parental occupation were recorded in all three surveys for each child. Height and weight were used to calculate BMI, weight was used to estimate BMR and parental occupation was used to determine social class. Comparing the macronutrient intakes in 2000 with 1980 and 1990, energy intakes (EI) fell in boys (to 8.45 MJ) and girls (to 7.60 MJ). This fall may, at least in part, be due to an increase in low energy reporting. For 1980, 1990 and 2000 the percentage of boys with EI:BMR below 1.1 was 6, 15 and 23 %, respectively; for girls, 3, 14 and 18 %, respectively. Percentage energy from fat was unchanged between 1980 and 1990 but fell to 35 % (about 76 g/d) in 2000, alongside a 3 % increase in percentage energy from starch (30 %). Percentage energy from non-milk extrinsic sugars remained above recommendations (16 %; about 82 g/d). The number of overweight and obese children increased from 11 % to 30 % between 1980 and 2000. Positive changes have occurred in the Northumbrian adolescent diet but social inequalities, reported in previous surveys, remain. PMID- 15333165 TI - Blood pressure responses in healthy older people to 50 g carbohydrate drinks with differing glycaemic effects. AB - The aim of the present study was to determine the effects on blood pressure response of 50 g carbohydrate drinks with differing glycaemic effects in ten healthy elderly subjects (age > 65 years; randomized crossover design). Systolic (SBP), diastolic (DBP) and mean arterial (MAP) blood pressure, heart rate and plasma glucose levels were determined following ingestion of equal volumes (379 ml) of water and 50 g carbohydrate drinks with differing reported glycaemic indices (GI) (surrogate marker for glycaemic effect): (1) low-GI: Apple & Cherry Juice; (2) intermediate-GI: Fanta Orange; (3) high-glucose. Glucose (SBP and DBP P < 0.001; MAP P = 0.005) and Fanta Orange (SBP P = 0.005; DBP and MAP P < 0.001) ingestion caused a significant decrease in BP whilst blood pressure increased (SBP P = 0.008; MAP P = 0.005) from baseline following Apple & Cherry Juice ingestion. Water had no significant effect on postprandial blood pressure. Fanta Orange and Apple & Cherry Juice caused similar (P = 0.679) glycaemic effects, which were significantly greater than water, but lower than glucose (P < 0.001). There was no significant correlation between the glycaemic effect of the carbohydrate drinks and there was no change in blood pressure from baseline (SBP r - 0.123, P = 0.509; DBP r - 0.051, P = 0.784; MAP r - 0.069, P = 0.712). Apple & Cherry Juice and Fanta Orange had similar glycaemic effects, but differing effects on blood pressure. Therefore, it is unlikely that the glycaemic effect of a drink can be used to predict the subsequent cardiovascular response. PMID- 15333166 TI - Attitudes towards alcohol of emergency department doctors trained in the detection of alcohol misuse. AB - INTRODUCTION: Alcohol misuse creates an immense burden for society, with problem drinkers too often constituting a neglected group. The Paddington Alcohol Test (PAT) is a useful screening tool in emergency departments. METHODS: Using a questionnaire, we assessed the attitudes of 127 emergency department junior doctors over 5 years to misuse detection using the PAT, in a centre with a well defined protocol for detection and referral. RESULTS: The majority (99%) thought early detection important, and the emergency department an appropriate place for screening (98%). Most thought that treatment could be successful (98%), and the PAT a useful instrument for early detection (87%). However, 63% reported that they misuse alcohol at least once a month and 30% once or more a week. DISCUSSION AND CONCLUSIONS: Junior doctors trained in the detection of alcohol misuse have a very positive view of this work. However, this professional insight is in marked contrast to their personal misuse of alcohol. This paradox reflects the entrenched culture of alcohol use in the medical profession, perhaps learnt at medical school. PMID- 15333167 TI - Artificial intelligence in medicine. AB - INTRODUCTION: Artificial intelligence is a branch of computer science capable of analysing complex medical data. Their potential to exploit meaningful relationship with in a data set can be used in the diagnosis, treatment and predicting outcome in many clinical scenarios. METHODS: Medline and internet searches were carried out using the keywords 'artificial intelligence' and 'neural networks (computer)'. Further references were obtained by cross referencing from key articles. An overview of different artificial intelligent techniques is presented in this paper along with the review of important clinical applications. RESULTS: The proficiency of artificial intelligent techniques has been explored in almost every field of medicine. Artificial neural network was the most commonly used analytical tool whilst other artificial intelligent techniques such as fuzzy expert systems, evolutionary computation and hybrid intelligent systems have all been used in different clinical settings. DISCUSSION: Artificial intelligence techniques have the potential to be applied in almost every field of medicine. There is need for further clinical trials which are appropriately designed before these emergent techniques find application in the real clinical setting. PMID- 15333168 TI - Consent in orthopaedic surgery. AB - AIM: To investigate whether the guidelines set out by the UK Department of Health on informed consent are being followed nationally in orthopaedic surgery. METHODS: A postal questionnaire of UK orthopaedic consultants was undertaken asking about consenting procedures for an elective and a trauma situation. RESULTS: In 53 of 110 cases, the most junior member of the team takes consent, and patients are not being warned about specific complications and risks associated with surgery. CONCLUSIONS: The guidelines issued by the Department of Health are not being adhered to, and the consenting doctor needs to be aware of the medical and legal responsibilities in taking informed consent. PMID- 15333169 TI - Reducing missing fracture clinic radiographs by entrusting them to patients. AB - BACKGROUND: Missing radiographs in fracture clinics may compromise fracture management and lead to inappropriate use of clerical resources. METHODS: We prospectively compared the number of missing radiographs in two hospitals over a period of two months. In hospital A the radiographs were retained and in hospital B they were entrusted to the patients. RESULTS: At the completion of the study, entrusting patients with their radiographs resulted in statistically less radiographs missing from the clinic. PMID- 15333170 TI - Treatment of osteoporosis: missed opportunities in the hospital fracture clinic. AB - AIM: To identify what proportion of patients who, having sustained an initial distal radial fragility fracture and a subsequent femoral neck fracture, had had their osteoporosis addressed in the interval between the two events. PATIENTS AND METHODS: The hospital electronic information system was used to identify all patients aged over 50 years treated for a distal radial fracture, in our fracture clinic, between 1995-2000. In addition all patients admitted to our hospital, during the same time period, with a femoral neck fracture were identified. RESULTS: A study cohort of 74 patients were identified. The proportion of patients who received investigation of, or treatment for, osteoporosis between their wrist and femoral neck fracture was 8% whereas 84% did not. CONCLUSIONS: Current mechanisms for identifying patients with osteoporosis before they sustain a femoral neck fracture are inadequate. PMID- 15333171 TI - Annual review of fractured neck of femur mortality rates: is this a true picture? AB - BACKGROUND: Annual fractured neck of femur death rates within a region are used by the Department of Health (DoH) as a clinical performance indicator of hospital trusts. Presently trusts and DoH publish performance figures annually but there is significant variability year to year. OBJECTIVE: Does a 5-year mortality review reflect a truer picture? METHODS: Hospital information services were used to provide the details for all patients who died within 30 days of admission. These were analysed to determine the true fractured neck of femur mortality. RESULTS: The variations in fractured neck of femur mortality rates between years within the same and between different trusts shows that annual scrutiny is an inappropriate, often skewed, analysis of performance. CONCLUSIONS: A 5-year mortality review would seem more appropriate to visualise true trends and should be performed to remove anomalous results. PMID- 15333172 TI - William Adams and the spine of Gideon Algernon Mantell. PMID- 15333173 TI - Is discharge home with drains after breast surgery producing satisfactory outcomes? AB - AIM: To assess whether discharging patients after breast cancer surgery with drains in situ and specialist nurse support is producing satisfactory outcomes. PATIENTS AND METHODS: A retrospective study assessing outcome subjectively and objectively in patients treated at the Breast Unit. The full population of patients (124) operated for breast cancer and discharged with drains in situ, comprising 84 mastectomies and 40 wide local excisions with axillary clearance (WLE). Patients were asked to complete a postal questionnaire, and complication and activity data were analysed. RESULTS: The questionnaire response rate was 60%. Only 10% of patients felt the care received at home was 'worse' than that received in hospital. Over a quarter of locally-conservative surgery patients described not being informed of the plan to discharge them with a drain in situ compared to only 9% of mastectomy patients. Only 6% of patients felt 'confident' going home early from hospital. On their first day at home, two-thirds of patients admitted feeling anxious and this was halved by the second day. Lack of confidence emptying drains was significantly worse in older patients. Complication rates were low and mean bed-days saved per patient was 4.7 days (SD 2.3). CONCLUSIONS: Overall patient satisfaction, hospital bed-days saved and low complication rates showed the efficacy of this early discharge policy. There were specific areas needing improvement. Patients should be more clearly informed of the early discharge plan. Greater attention should be paid to allaying patient anxiety during the first day at home, especially in older patients. PMID- 15333174 TI - Delayed cholecystectomy for gallstone pancreatitis: re-admissions and outcomes. AB - OBJECTIVE: Timing of cholecystectomy after gallstone pancreatitis and use of pre operative cholangiography varies considerably between surgeons. We examined outcomes in a district general hospital where most patients underwent delayed cholecystectomy following pre-operative cholangiography. METHODS: A retrospective review of admissions with gallstone pancreatitis over a 5-year period was conducted. RESULTS: A total of 77 patients with gallstone pancreatitis were identified of whom 58 underwent laparoscopic cholecystectomy (LC) at a median of 67.5 days after index admission. Of these patients, 21% had unplanned re admission while awaiting LC rising to 25% of those who waited for more than 4 weeks. Surgery at 4 weeks would have been associated with a 6% re-admission rate. Re-admissions were due to pancreatitis (4 cases), cholecystitis (3 cases), biliary colic (4 cases) and pseudocyst (1 case). In all, 49 patients had pre operative cholangiography and 13 had pre-operative endoscopic extraction of stones from the common bile duct. CONCLUSIONS: Delay of LC for greater than 4 weeks after gallstone pancreatitis is associated with a high, unplanned re admission rate, even with liberal use of pre-operative cholangiography. PMID- 15333175 TI - Open mesh repair of incisional hernias with significant loss of domain. AB - BACKGROUND: Incisional hernias develop in up to 13% of laparotomy incisions: the most difficult to repair are complex, multiply recurrent hernias with significant loss of domain (>15-20% of the abdominal contents). METHODS: Retrospective analysis by standard proforma of a series of 52 patients operated on at a single institution between 1996 and 2002. All patients received pre-operative CT and anaesthetic assessment. Patients with significant tissue loss were assessed by a plastic surgeon. Cardiorespiratory status was optimised and trophic skin ulcers treated before operation. RESULTS: Sublay repair was applied in 33 patients, onlay in 16 patients, one patient received inlay repair and two patients the Ramirez abdominoplasty. Additional procedures of stoma closure, muscle flap or abdominoplasty were carried out in 7 patients. Complications occurred in 18 (34.6%) patients, 5 of whom required further surgery for haematoma, infection or fistulisation. One patient died from pulmonary embolism after postoperative complications. Three recurrences were apparent after follow-up of 6 months to 6 years. CONCLUSIONS: Complex incisional hernias are a challenging surgical problem. Careful patient selection and surgical technique with a team involving anaesthetists and plastic surgeons is required. Post-operative management may require facilities in HDU and ITU. Clinical trials are required to identify techniques and materials which give the best results. PMID- 15333176 TI - Flexible ureterorenoscopy: prospective analysis of the Guy's experience. AB - OBJECTIVE: To report our outcomes with small diameter, actively deflectable flexible ureterorenoscopy from a prospective database. PATIENTS AND METHODS: 114 flexible ureterorenoscopies were performed in 105 patients (mean age, 49.5 years; range, 19-85 years; 71 males, 34 females) over a 9-month period. Of these, 101 were for refractory stones following failed ESWL and 13 for diagnostic reasons. An Olympus URF P3 flexible ureteroscope with pressure irrigation was used. Electrohydraulic lithotripsy was used to fragment stones and the fragments were retrieved with Graspit, triradiate graspers or tipless baskets. RESULTS: Stents had previously been placed in 53% and dilatation of the ureteric orifice was necessary in 15%. In the stone group, the median operating time was 55 min (range, 15-210 min) and the median screening time 2.2 min (range, 0.3-9.1 min). Success was defined as complete stone clearance or good fragmentation to 2 mm or less. Overall success in this group was 72.3%. There was no statistically significant difference between lower and other calyces (P=0.83 Chi-square test). Successful outcome was achieved in 72% for stone size 10 mm or less, 80% for 11 20 mm and 50% for greater than 20 mm. Two or more procedures were needed in 8 patients. In the diagnostic group, the median operating time was 45 min (range, 20-60 min) and the median screening time 2 min (range, 0.3-8.3 min). The majority were for upper tract filling defects. Access and successful diagnosis was achieved in all cases. The major complication rate was 2.6%. The ureteroscope needed repair once during this series. CONCLUSIONS: Flexible ureterorenoscopy is an effective diagnostic and therapeutic tool in a select group of patients. It should be considered for ESWL-resistant upper tract stones but the results are poor in stones larger than 20 mm and percutaneous nephrolithotomy may be a better option in these patients. PMID- 15333178 TI - Endovascular repair (EVAR) of abdominal aortic aneurysms: the case for. PMID- 15333179 TI - The educational evaluation of General Professional Practice of Surgery (GPPS). AB - The authors conclude that GPPS presents the possibility of enhanced learning from limited opportunities, but only if all parties are fully able to embrace the educational aims of GPPS and follow through its structured and coherent design for learning. PMID- 15333180 TI - Spontaneous passage of bile duct stones: frequency of occurrence and relation to clinical presentation. PMID- 15333182 TI - Amputations at St Thomas' hospital 1862-1869. PMID- 15333183 TI - Ultrasound diagnosis of round ligament varicosities mimicking inguinal hernias in pregnancy. PMID- 15333184 TI - Liver resection for colorectal metastases. PMID- 15333185 TI - Referral guidelines for colorectal cancer--do they work? PMID- 15333186 TI - Matador versus taurus: bull gore injury. PMID- 15333187 TI - Detection and quantitation of normal and variant haemoglobins: an analytical review. AB - As well as six 'normal' haemoglobins that occur at various stages of development, more than 800 abnormal or variant haemoglobins have been described. Many of these variant haemoglobins have no significant clinical consequences apart from causing confusion to clinicians and in laboratories; however, some of the variant haemoglobins result in major morbidity or mortality. The laboratory challenge is to detect these clinically significant haemoglobins and to identify them with sufficient accuracy for clinical purposes, as well as to quantitate both these and the 'normal' haemoglobins. The techniques used to detect and quantitate these haemoglobins in routine service laboratories are discussed in detail. Methods used by referral laboratories, such as mass spectrometry and DNA analysis, are briefly discussed. Haemoglobin analysis is most often undertaken as part of neonatal, antenatal or pre-anaesthetic screening; these programmes are reviewed, together with possible changes to neonatal screening and antenatal screening that may occur as part of the NHS National Plan. PMID- 15333188 TI - Evidence for the clinical use of tumour markers. AB - Testing for tumour markers should only be performed if it results in a better patient outcome, increased quality of life or reduced overall cost of care. Ideally, the clinical value of a tumour marker should be validated in a large prospective study or a meta-analysis of small-scale retrospective/prospective studies (i.e. a level 1 evidence study) prior to routine use. Markers that have been validated in such a level 1 evidence study include carcinoembryonic antigen in the surveillance of patients with diagnosed colorectal cancer, alphafetoprotein, human chorionic gonadotrophin and lactate dehydrogenase for evaluating prognosis in patients non-seminomatous germ cell tumours, CA 125 for monitoring therapy in patients with ovarian cancer, oestrogen receptors for predicting response to hormone therapy in breast cancer, HER-2 for predicting response to trastuzumab in patients with advanced breast cancer and urokinase plasminogen activator/plasminogen activator inhibitor type 1 for determining prognosis in breast cancer. Although currently in widespread use, the value of prostate-specific antigen in screening for prostate cancer has yet to be validated in a large prospective randomized trial. PMID- 15333189 TI - An audit of tumour marker requests in Northern Ireland. AB - BACKGROUND: An audit of tumour marker requests was carried out in Northern Ireland. The aims were to establish the extent of inappropriate requesting, to effect change in request practice if required, and to standardize laboratory reports for tumour markers. METHODS: A baseline audit was conducted to investigate the appropriateness of tumour marker requesting patterns in Northern Ireland. Information booklets containing guidelines for tumour markers were then distributed to all clinicians and a post-intervention audit was carried out. RESULTS: At baseline, 80% of requests for tumour markers were appropriate for the source of the tumour; however, 54% of clinicians used tumour markers to screen for malignancy and there was a low index of suspicion in 35% of these. Requests for more than two tumour markers were received for 5% of request forms. The post intervention survey showed no change in requesting practice. Comments were returned by requestors for 55% of questionnaires. They appeared to be appropriate in 72% of cases. CONCLUSIONS: There is a low level of inappropriate requesting of tumour markers in Northern Ireland. Audit and feedback combined with circulation of guidelines were unsuccessful interventions in improving the appropriateness of request practice for these tests. PMID- 15333190 TI - Objective determination of appropriate reporting intervals. AB - BACKGROUND: The reporting interval is the incremental value chosen in reporting analyte concentration. Reporting intervals for different analytes are often inappropriately narrow, when analytical imprecision and biological variability are considered. METHODS: We have used statistical techniques to determine intervals for individual analytes at which there is 50% or 95% confidence that two results are analytically different, and compared these with the reporting intervals in use for a range of general chemistry analytes and analytes usually measured by immunoassay. RESULTS: No analytes met the criteria for 95% confidence that the results are analytically different. Even at the 50% confidence level, 24 of 46 analytes failed at all concentrations examined. For some analytes, particularly hormones at high concentration, the reporting interval increment should be increased by a factor of at least ten. CONCLUSIONS: The majority of analytes are inappropriately reported when analytical precision alone is considered. The concept of the 'uncertainty of measurement' has not been adequately addressed. A consensus should be reached and implemented on appropriate reporting intervals for all analytes. PMID- 15333191 TI - Diagnostic value of heart fatty acid binding protein and myoglobin in patients admitted with chest pain. AB - INTRODUCTION: Patients presenting with chest pain do so at varying times following its onset. In addition, their histories can be unreliable, making the interpretation of time-dependent biochemical markers difficult. Patients are therefore subsequently managed according to measurements of specific markers, such as troponin, taken many hours following their admission. Earlier markers such as myoglobin and heart fatty acid binding protein (hFABP) may provide information earlier; however, the true role of such markers remains unclear when applied to real life patient cohorts. We aimed to evaluate and compare the diagnostic accuracy of the early markers hFABP and myoglobin (in terms of area under ROC curves, sensitivity and specificity) to troponin I (cTnI) in a pragmatic group of patients presenting with chest pain. METHODS: 302 patients presenting with chest pain had blood samples withdrawn both on admission and 12 h later. These were analysed for cTnI, myoglobin and hFABP. Standard clinical and electrocardiographic data were also obtained. Only the cTnI results were reported back to the clinicians and hence only they would have affected clinical care. Patients were grouped clinically according to both the American College of Cardiology/European Society of Cardiology new definition of myocardial infarction (MI) and by clinicians' diagnosis of MI. RESULTS: Median time from onset of symptoms to admission was 5.0 h (IQR 3.0-12 h). Greater ROC curve areas for the diagnosis of MI by both sets of criteria were obtained for cTnI over both myoglobin and hFABP. ROC curve areas for myoglobin and hFABP were also no better than that for cTnI in those patients presenting less than 4 h after the onset of chest pain. Both myoglobin and hFABP also performed poorly in those patients who were cTnI negative on admission. CONCLUSION: Myoglobin and hFABP provide little clinical value when measured on admission in patients presenting with chest pain. PMID- 15333192 TI - A prospective randomized controlled trial of point-of-care testing on the coronary care unit. AB - BACKGROUND: We report the results of a prospective randomized controlled trial comparing point-of-care testing (POCT) with central laboratory testing (CLT) in a six-bed coronary care unit in a district general hospital. METHODS: 263 consecutive admissions with chest pain and suspected acute coronary syndrome were randomized to measurement of cardiac troponin T by POCT or CLT only. Patient management was according to a pre-specified protocol utilizing clinical features, electrocardiographic changes and cardiac biomarkers (creatine kinase and cardiac troponin T) to define management. Outcome measures were diagnostic accuracy compared with CLT as 'gold standard', result turnaround time, mortality and length of stay in all patients and those with a protocol-driven early discharge policy. RESULTS: Diagnostic accuracy and mortality was equivalent in the POCT and CLT arm. Overall there was no difference in length of stay. In the pre-specified early discharge group (n = 64) there was a significant reduction in median length of non-coronary care unit stay (145.3 h versus 79.5 h) and overall hospital stay (209.3 h versus 149.9 h) in those randomized to POCT. CONCLUSION: A combination of rapid biochemical diagnosis and structured decision making reduces length of hospital stay. PMID- 15333193 TI - Vitamin D deficiency masking primary hyperparathyroidism. AB - Vitamin D deficiency and primary hyperparathyroidism (PHPT) are relatively common disorders. The coexistence of these conditions should be considered, as depletion of vitamin D may alter the clinical expression of autonomous parathyroid disease. We report details of a vitamin D deficient patient in whom replacement therapy led to the unmasking of occult PHPT. PMID- 15333194 TI - Morning salivary cortisol versus short Synacthen test as a test of adrenal suppression. AB - BACKGROUND: The short Synacthen test (SST) is the most commonly used test for the assessment of adrenal suppression. We investigated the potential of a simpler and more cost-effective procedure [morning salivary cortisol (MSC)] as an outpatient screening tool to detect adrenal suppression in patients using topical intranasal corticosteroids for rhinosinusitis. METHOD: Forty-eight patients who were using topical corticosteroids underwent adrenal function assessment by way of SST and MSC measurement. RESULTS: Sixteen of the 48 patients had impaired MSCs. Of these 16 patients, 15 had an impaired SST (sensitivity 100%) and one had a normal SST. All patients with normal MSCs also had normal SSTs (specificity 97%). CONCLUSION: The morning salivary cortisol measurement is a useful screening tool for adrenal suppression in this setting. PMID- 15333195 TI - Point-of-care test to monitor adherence to anti-tuberculous treatment. AB - BACKGROUND: In the fight against the global tuberculosis epidemic, it is essential to ensure that patients adhere to the treatment prescribed. As the treatment is given for a minimum of 6 months it is common for patients not to take their drugs regularly. Strategies are therefore needed to assess adherence to treatment. One established method is to examine the patient's urine for the presence of drug metabolites. A rapid point-of-care test would overcome some of the drawbacks associated with currently available methods. METHOD: A rapid, safe point-of-care test for isoniazid metabolites (IsoScreen, Surescreen Diagnostics Limited, Derby, UK) has been developed and used to help assess adherence to treatment in a busy clinic for tuberculosis patients in South London. RESULTS: Urine samples were examined from 191 patients receiving isoniazid, usually in combination with rifampicin and other anti-tuberculous drugs. Isoscreen was positive in 93.2% of patients, suggesting that 6.8% might be poorly adhering to treatment. By contrast, examining the same urine samples for evidence of rifampicin ingestion gave positive results in only 43.5%, due to the fact that this test is only positive for a few hours after drug ingestion. CONCLUSION: IsoScreen has been shown to provide a rapid and safe point-of-care test, which contributes to the detection of non-adherence in patients with tuberculosis. PMID- 15333196 TI - Iatrogenic copper deficiency causing anaemia and neutropenia. AB - A 79-year-old woman presented with an obstructed femoral hernia and had a wedge resection of the small intestine. Post-operatively she developed wound dehiscence and spent 3 days in the adult critical care unit. Good recovery followed and she was allowed home after 2 months in hospital. Six months later she presented with anaemia, neutropenia and a very low serum copper concentration. Review of her notes revealed that she had been given oral zinc therapy while in the critical care unit and this treatment had been continued on discharge from hospital. Serum copper, haemoglobin and white cell count recovered after oral zinc was discontinued. Oral zinc treatment can lead to symptomatic copper deficiency in susceptible patients. PMID- 15333197 TI - Iatrogenic copper deficiency following information and drugs obtained over the Internet. AB - We report the case of a 56-year-old woman with a 7-year history of metastatic cancer who presented with severe copper deficiency following self-treatment with the copper-chelating agent tetrathiomolybdate. This compound was used with the aim of inhibiting tumour angiogenesis and was obtained from the USA by placing an order on the internet. The patient exhibited severe neutropenia as her serum copper concentration fell from 19.8 micromol/L to 3.3 micromol/L and her caeruloplasmin concentration from 35 mg/dL to 4 mg/dL. PMID- 15333198 TI - Stability of cystatin C in urine. PMID- 15333199 TI - Dimercaptosuccinic acid loading test for assessing mercury burden in healthy individuals. PMID- 15333200 TI - Dimercaptosuccinic acid loading test for assessing mercury burden in healthy individuals. PMID- 15333205 TI - Different predictions by the minimum variance and minimum torque-change models on the skewness of movement velocity profiles. AB - We investigated the differences between two well-known optimization principles for understanding movement planning: the minimum variance (MV) model of Harris and Wolpert (1998) and the minimum torque change (MTC) model of Uno, Kawato, and Suzuki (1989). Both models accurately describe the properties of human reaching movements in ordinary situations (e.g., nearly straight paths and bell-shaped velocity profiles). However, we found that the two models can make very different predictions when external forces are applied or when the movement duration is increased. We considered a second-order linear system for the motor plant that has been used previously to simulate eye movements and single-joint arm movements and were able to derive analytical solutions based on the MV and MTC assumptions. With the linear plant, the MTC model predicts that the movement velocity profile should always be symmetrical, independent of the external forces and movement duration. In contrast, the MV model strongly depends on the movement duration and the system's degree of stability; the latter in turn depends on the total forces. The MV model thus predicts a skewed velocity profile under many circumstances. For example, it predicts that the peak location should be skewed toward the end of the movement when the movement duration is increased in the absence of any elastic force. It also predicts that with appropriate viscous and elastic forces applied to increase system stability, the velocity profile should be skewed toward the beginning of the movement. The velocity profiles predicted by the MV model can even show oscillations when the plant becomes highly oscillatory. Our analytical and simulation results suggest specific experiments for testing the validity of the two models. PMID- 15333206 TI - Disambiguating visual motion through contextual feedback modulation. AB - Motion of an extended boundary can be measured locally by neurons only orthogonal to its orientation (aperture problem) while this ambiguity is resolved for localized image features, such as corners or nonocclusion junctions. The integration of local motion signals sampled along the outline of a moving form reveals the object velocity. We propose a new model of V1-MT feedforward and feedback processing in which localized V1 motion signals are integrated along the feedforward path by model MT cells. Top-down feedback from MT cells in turn emphasizes model V1 motion activities of matching velocity by excitatory modulation and thus realizes an attentional gating mechanism. The model dynamics implement a guided filling-in process to disambiguate motion signals through biased on-center, off-surround competition. Our model makes predictions concerning the time course of cells in area MT and V1 and the disambiguation process of activity patterns in these areas and serves as a means to link physiological mechanisms with perceptual behavior. We further demonstrate that our model also successfully processes natural image sequences. PMID- 15333204 TI - Understanding the cortical specialization for horizontal disparity. AB - Because the eyes are displaced horizontally, binocular vision is inherently anisotropic. Recent experimental work has uncovered evidence of this anisotropy in primary visual cortex (V1): neurons respond over a wider range of horizontal than vertical disparity, regardless of their orientation tuning. This probably reflects the horizontally elongated distribution of two-dimensional disparity experienced by the visual system, but it conflicts with all existing models of disparity selectivity, in which the relative response range to vertical and horizontal disparities is determined by the preferred orientation. Potentially, this discrepancy could require us to abandon the widely held view that processing in V1 neurons is initially linear. Here, we show that these new experimental data can be reconciled with an initial linear stage; we present two physiologically plausible ways of extending existing models to achieve this. First, we allow neurons to receive input from multiple binocular subunits with different position disparities (previous models have assumed all subunits have identical position and phase disparity). Then we incorporate a form of divisive normalization, which has successfully explained many response properties of V1 neurons but has not previously been incorporated into a model of disparity selectivity. We show that either of these mechanisms decouples disparity tuning from orientation tuning and discuss how the models could be tested experimentally. This represents the first explanation of how the cortical specialization for horizontal disparity may be achieved. PMID- 15333207 TI - Exact solution for the optimal neuronal layout problem. AB - Evolution perfected brain design by maximizing its functionality while minimizing costs associated with building and maintaining it. Assumption that brain functionality is specified by neuronal connectivity, implemented by costly biological wiring, leads to the following optimal design problem. For a given neuronal connectivity, find a spatial layout of neurons that minimizes the wiring cost. Unfortunately, this problem is difficult to solve because the number of possible layouts is often astronomically large. We argue that the wiring cost may scale as wire length squared, reducing the optimal layout problem to a constrained minimization of a quadratic form. For biologically plausible constraints, this problem has exact analytical solutions, which give reasonable approximations to actual layouts in the brain. These solutions make the inverse problem of inferring neuronal connectivity from neuronal layout more tractable. PMID- 15333208 TI - Decoding a temporal population code. AB - Encoding of sensory events in internal states of the brain requires that this information can be decoded by other neural structures. The encoding of sensory events can involve both the spatial organization of neuronal activity and its temporal dynamics. Here we investigate the issue of decoding in the context of a recently proposed encoding scheme: the temporal population code. In this code, the geometric properties of visual stimuli become encoded into the temporal response characteristics of the summed activities of a population of cortical neurons. For its decoding, we evaluate a model based on the structure and dynamics of cortical microcircuits that is proposed for computations on continuous temporal streams: the liquid state machine. Employing the original proposal of the decoding network results in a moderate performance. Our analysis shows that the temporal mixing of subsequent stimuli results in a joint representation that compromises their classification. To overcome this problem, we investigate a number of initialization strategies. Whereas we observe that a deterministically initialized network results in the best performance, we find that in case the network is never reset, that is, it continuously processes the sequence of stimuli, the classification performance is greatly hampered by the mixing of information from past and present stimuli. We conclude that this problem of the mixing of temporally segregated information is not specific to this particular decoding model but relates to a general problem that any circuit that processes continuous streams of temporal information needs to solve. Furthermore, as both the encoding and decoding components of our network have been independently proposed as models of the cerebral cortex, our results suggest that the brain could solve the problem of temporal mixing by applying reset signals at stimulus onset, leading to a temporal segmentation of a continuous input stream. PMID- 15333209 TI - Minimal models of adapted neuronal response to in vivo-like input currents. AB - Rate models are often used to study the behavior of large networks of spiking neurons. Here we propose a procedure to derive rate models that take into account the fluctuations of the input current and firing-rate adaptation, two ubiquitous features in the central nervous system that have been previously overlooked in constructing rate models. The procedure is general and applies to any model of firing unit. As examples, we apply it to the leaky integrate-and-fire (IF) neuron, the leaky IF neuron with reversal potentials, and to the quadratic IF neuron. Two mechanisms of adaptation are considered, one due to an afterhyperpolarization current and the other to an adapting threshold for spike emission. The parameters of these simple models can be tuned to match experimental data obtained from neocortical pyramidal neurons. Finally, we show how the stationary model can be used to predict the time-varying activity of a large population of adapting neurons. PMID- 15333210 TI - Including long-range dependence in integrate-and-fire models of the high interspike-interval variability of cortical neurons. AB - Many different types of integrate-and-fire models have been designed in order to explain how it is possible for a cortical neuron to integrate over many independent inputs while still producing highly variable spike trains. Within this context, the variability of spike trains has been almost exclusively measured using the coefficient of variation of interspike intervals. However, another important statistical property that has been found in cortical spike trains and is closely associated with their high firing variability is long-range dependence. We investigate the conditions, if any, under which such models produce output spike trains with both interspike-interval variability and long range dependence similar to those that have previously been measured from actual cortical neurons. We first show analytically that a large class of high variability integrate-and-fire models is incapable of producing such outputs based on the fact that their output spike trains are always mathematically equivalent to renewal processes. This class of models subsumes a majority of previously published models, including those that use excitation-inhibition balance, correlated inputs, partial reset, or nonlinear leakage to produce outputs with high variability. Next, we study integrate-and-fire models that have (nonPoissonian) renewal point process inputs instead of the Poisson point process inputs used in the preceding class of models. The confluence of our analytical and simulation results implies that the renewal-input model is capable of producing high variability and long-range dependence comparable to that seen in spike trains recorded from cortical neurons, but only if the interspike intervals of the inputs have infinite variance, a physiologically unrealistic condition. Finally, we suggest a new integrate-and-fire model that does not suffer any of the previously mentioned shortcomings. By analyzing simulation results for this model, we show that it is capable of producing output spike trains with interspike-interval variability and long-range dependence that match empirical data from cortical spike trains. This model is similar to the other models in this study, except that its inputs are fractional-gaussian-noise-driven Poisson processes rather than renewal point processes. In addition to this model's success in producing realistic output spike trains, its inputs have long-range dependence similar to that found in most subcortical neurons in sensory pathways, including the inputs to cortex. Analysis of output spike trains from simulations of this model also shows that a tight balance between the amounts of excitation and inhibition at the inputs to cortical neurons is not necessary for high interspike-interval variability at their outputs. Furthermore, in our analysis of this model, we show that the superposition of many fractional-gaussian-noise driven Poisson processes does not approximate a Poisson process, which challenges the common assumption that the total effect of a large number of inputs on a neuron is well represented by a Poisson process. PMID- 15333211 TI - Learning eigenfunctions links spectral embedding and kernel PCA. AB - In this letter, we show a direct relation between spectral embedding methods and kernel principal components analysis and how both are special cases of a more general learning problem: learning the principal eigenfunctions of an operator defined from a kernel and the unknown data-generating density. Whereas spectral embedding methods provided only coordinates for the training points, the analysis justifies a simple extension to out-of-sample examples (the Nystrom formula) for multidimensional scaling (MDS), spectral clustering, Laplacian eigenmaps, locally linear embedding (LLE), and Isomap. The analysis provides, for all such spectral embedding methods, the definition of a loss function, whose empirical average is minimized by the traditional algorithms. The asymptotic expected value of that loss defines a generalization performance and clarifies what these algorithms are trying to learn. Experiments with LLE, Isomap, spectral clustering, and MDS show that this out-of-sample embedding formula generalizes well, with a level of error comparable to the effect of small perturbations of the training set on the embedding. PMID- 15333212 TI - Transurethral ethanol ablation of prostate. AB - Transurethral ethanol ablation of the prostate (TEAP) has emerged among the treatment alternatives to transurethral resection of prostate as a promising minimally invasive therapy that can be performed on an outpatient basis with fewer complications. It was introduced approximately 5 years ago, and to date, 12 month results are encouraging. We herein review the procedure and outcomes of TEAP. PMID- 15333213 TI - The birth of modern ureteroscopy: the Albona Jaybis story. PMID- 15333214 TI - Results of shockwave lithotripsy for pediatric urolithiasis. AB - BACKGROUND AND PURPOSE: Shockwave lithotripsy (SWL) is widely practiced in the management of pediatric urolithiasis. However, the efficacy, need for ancillary procedures, and treatment-related complications are not as clearly defined as in the adult population. We reviewed the outcomes of SWL in the pediatric population at our lithotripsy unit. PATIENTS AND METHODS: A retrospective review of all patients /=4 mm on presentation were 26% more likely to fail observation than patients with smaller solitary calculi (P = 0.012). Stone growth was less common in those with upper-pole and middle-pole stones (47% v 61%) for lower-pole stones; P = 0.002). Urine uric acid concentration correlated positively with the rate of stone growth (P = 0.05). Likewise, serum uric acid concentration predicted stone growth (odds ratio = 3.6). CONCLUSIONS: Stratification of risk of progression according to presenting stone size, location, and composition may facilitate discussions with the patient regarding the alternatives of observation versus intervention. Specifically, small non-uric acid calculi in the upper pole may be most amenable to observation. PMID- 15333217 TI - Virtual endoscopy in renal artery stenosis: an innovative approach for diagnosis and follow-up. AB - PURPOSE: We investigated the utility of virtual endoscopy (VE) as a diagnostic and follow-up tool in patients with renal artery stenosis, especially as a means of defining vascular patency. PATIENTS AND METHODS: We performed VE in 24 patients with ostial atherosclerotic renal artery stenosis and correlated the results with those of conventional angiography. The patients were treated successfully by placement of metal stents and conventional catheter angiography and VE for patency assessment 6 and 12 months after stent insertion. RESULTS: In all patients, the stenotic segment was identified, and VE findings were concordant with those of angiography. The average degree of stenosis was estimated to be 70% +/- 20% when angiography was used and 62% +/- 15% when VE was used. After metal stent insertion, the 12-month patency rate was 83.3% (20 patients). Angiography and VE findings remained concordant during the follow-up period, but VE provided more information beyond the stenotic segment, allowing examination of the arterial lumen both cephalad and caudal to the point of obstruction. CONCLUSION: Virtual endoscopy provided a more dynamic, direct, minimally invasive approach that was equal to or better than angiography for both the verification of the vascular stenosis and the evaluation of the arterial lumen. PMID- 15333218 TI - Renal capsular block: a novel method for performing percutaneous nephrolithotomy under local anesthesia. AB - Most of the pain of percutaneous nephrolithotomy is caused by dilatation of the renal capsule and parenchymal tract. We evolved a technique to block renal capsular sensations by infiltration of lignocaine (lidocaine) at the site of renal entry, permitting the procedure to be done with local anesthesia. PMID- 15333219 TI - Cauterization of access tract for nephrostomy tube-free percutaneous nephrolithotomy. AB - BACKGROUND AND PURPOSE: Percutaneous nephrolithotomy (PCNL) is now a popular method for removal of renal and ureteral stones. Placement of a nephrostomy tube after the completion of PCNL has been considered a standard procedure by most urologists, but some authors have recently challenged this practice. Bleeding is one of the most prevalent problems after nephrostomy tube-free percutaneous renal surgery. To diminish the possibility of postoperative bleeding, we cauterized the PCNL tract to make it bloodless. The efficacy and safety of this procedure were reviewed in this study. PATIENTS AND METHODS: From March 2001 to March 2003, 51 patients underwent PCNL with a one-stage procedure and a single access tract. The stone size ranged from 1.0 to 7.0 cm (mean 2.7 +/- 1.4 cm). A holmium:YAG laser and pneumatic lithotripter were used. After stone extraction, a 6F double-J catheter was inserted antegrade. The access tract was checked, and the bleeding points were cauterized. No nephrostomy tube was inserted, but a Penrose drain was left overnight. Perforation of the collecting system was not a contraindication to tubeless PCNL. RESULTS: The stone-free rate was 80.4%, including five patients with complete staghorn stones. Twenty-one patients required postoperative analgesics. Only one patient had urine leakage for longer than 24 hours. Transient low fever was noted in five patients, but no patient experienced severe urinary tract infection. Delayed hemorrhage (1 week after the operation) secondary to irritation by the double-J ureteral stent was noted in one patient. The average postoperative hospital stay was 2.2 days (range 1-3 days). No patient required a blood transfusion. No urinoma was noted on the postoperative ultrasound follow-up. CONCLUSION: Nephrostomy tube-free percutaneous renal surgery is a safe and effective procedure for selected patients with minimal hemorrhage after PCNL. Cauterization of tract bleeding points may make this modification a more secure procedure and make it suitable for more patients. PMID- 15333220 TI - Assessment of encrustations on polyurethane ureteral stents. AB - PURPOSE: To determine the composition and the extent of crystalline (and other) encrustation on ureteral catheters inserted under sterile conditions in stone formers, in comparison with catheters of the same type inserted in nonstone formers for the same time but for different clinical reasons. MATERIALS AND METHODS: Forty consecutive self-retained polyurethane pigtail ureteral catheters removed by cystoscopy between November 2000 and February 2002 were studied, 30 from stone formers and 10 from patients without stone histories. The mean dwelling time was 55 days for the stone formers and 79 days for the other patients. The encrustations were collected and analyzed with Fourier-transform infrared spectroscopy, powder X-ray diffraction, or both. The stones from nine of the patients were also subjected to the same spectroscopic analysis. Representative sections of the catheters were investigated by scanning electron microscopy and energy-dispersive X-ray analysis. RESULTS: The most common encrustation in stone formers was calcium oxalate monohydrate. In patients without stones, deposits of organic compounds were found consistently. The mean mass of encrustation of stone formers was larger (71.05 mg) than that of patients without stones (1 mg). CONCLUSIONS: Calcium oxalate is the predominant type of encrustation on ureteral catheters in stone formers. Prevention of heavy encrustation should be directed to therapeutic measures concerning calcium oxalate lithiasis and development of new materials by the medical industry that are less prone to encrustation. PMID- 15333221 TI - Laparoscopic retroperitoneal ureterolithotomy: initial experience and review of literature. AB - PURPOSE: To evaluate the place of retroperitoneal laparoscopic ureterolithotomy in the management of ureteral calculi through an initial experience and review of previously reported cases. PATIENTS AND METHODS: Laparoscopic ureterolithotomy was attempted in six patients (five men and one woman) with an age of 31 to 56 years (mean 41.5 years) who had large and impacted stones in the upper ureter. The stone size ranged from 18 to 40 mm (mean 25.7 mm). The retroperitoneal route was used in all cases. RESULTS: The stone was removed in all but one case. The operative time ranged from 120 to 180 minutes (mean 160 minutes). No intraoperative complications were encountered. Postoperative complications included prolonged urinary leak in one patient. On follow-up, all patients were stone free with decompression of the upper urinary tract. CONCLUSIONS: Laparoscopic ureterolithotomy is safe and effective and should be performed each time a ureterolithotomy is indicated. PMID- 15333222 TI - Hand-assisted laparoscopic heminephrectomy in horseshoe kidney. AB - BACKGROUND AND PURPOSE: Horseshoe kidneys are the most common renal fusion anomalies. In surgery on normal kidneys, the use of minimally invasive techniques has decreased morbidity and improved postoperative care. However, aberrant vasculature, abnormal kidney location, and the renal isthmus present technical challenges to the laparoscopic management of horseshoe kidneys. We describe a case of heminephrectomy using hand-assisted laparoscopic techniques. METHODS: A 35-year-old female patient with a horseshoe kidney was evaluated for recurrent nephrolithiasis and infections. Despite multiple previous percutaneous stone removal procedures, she continued to have symptomatic nephrolithiasis in the atrophic and obstructed right renal moiety. A transperitoneal hand-assisted laparoscopic heminephrectomy was performed on the right side. The aberrant vessels were identified and divided while the renal isthmus was divided using an endoscopic stapler. RESULTS: The operative time was 165 minutes, while the estimated blood loss was 200 mL. There were no intraoperative complications. Postoperative pain was minimal, and the hospital stay was 4 days. The patient remains asymptomatic 6 months after her partial nephrectomy, with stable renal function. CONCLUSION: Hand-assisted laparoscopic heminephrectomy provides a safe treatment option for patients with an atrophic, obstructed moiety in a horseshoe kidney. PMID- 15333223 TI - Acute recovery of pneumoperitoneum using argon gas. AB - BACKGROUND AND PURPOSE: The argon-beam coagulator (ABC) is widely used in laparoscopic surgery as a method of electrocoagulation. Argon gas possesses properties that make it suitable as an alternative for establishing pneumoperitoneum. We present a case in which an ABC was used to deliver argon gas urgently in order to salvage an acutely compromised pneumoperitoneum. METHODS: During a right partial nephrectomy, after the renal artery was clamped, a failure in the central CO(2) compressor compromised the pneumoperitoneum. Argon gas was delivered from an ABC at a flow rate of 4 L/min and a distance of 1 to 5 cm and directed toward the paranephric fat. RESULTS: Pneumoperitoneum was rapidly reestablished. Stable intra-abdominal pressure was maintained in the range of 14 to 20 mm Hg for 16 minutes until the original CO(2) supply was reestablished. The total warm ischemia time was 32 minutes. No hemodynamic changes were noted while using argon gas, and the procedure was completed successfully with an uneventful postoperative course. CONCLUSIONS: Argon gas delivery from an ABC can be used for emergency salvage of pneumoperitoneum in selected cases of acute CO(2) gas delivery failure and under strict intra-abdominal pressure monitoring. PMID- 15333224 TI - Overcoming the steep learning curve of laparoscopic radical prostatectomy: single surgeon experience. AB - PURPOSE: To evaluate the influence of intensive laparoscopic skills training and self-critical video review on the learning curve for laparoscopic radical prostatectomy (LRP). PATIENTS AND METHODS: The initial 40 patients who underwent a transperitoneal LRP (groups 1-4) and the subsequent 20 who underwent LRP by the extraperitoneal approach (group 5) were studied. Eight weeks prior to initiating the LRP program, intensive laparoscopic skills training at a minimally invasive surgery center was undertaken for an average of 4 hours per week. This self training was continued for 12 weeks into the program, with self-critical review of videotapes of each procedure. The groups were compared with respect to total operative time (ORT), anastomosis time, and blood loss. RESULTS: There were significant differences in the ORT and anastomosis times between each of the first two groups and the last two groups (P <0.001). The learning curve for ORT was overcome after approximately 35 cases, as there were no significant differences in ORT between group 3 and the subsequent groups. The anastomosis took longer to master, as significant time decreases were observed up to group 4, after which, the mean reached a plateau (group 4 v 5 P = NS). The differences in blood loss were not significant. Overall, there were 7 intraoperative (12.7%) and 8 postoperative (14.5%) complications. CONCLUSIONS: The use of similar facilities and training tools can help overcome the steep learning curve of LRP. Longer follow-up is needed to assess these means of attaining better functional results after LRP. PMID- 15333225 TI - Anatomic guide for port placement for daVinci robotic radical prostatectomy. AB - BACKGROUND AND PURPOSE: At present, robotic arm port placement for daVinci trade mark robot-assisted laparoscopic radical prostatectomy is based on the umbilicus. However, the robotic arm has a maximum manufactured required working distance of 25 cm. Accordingly, normal variability of patient height, weight, and umbilical location can leave the working arms too short to reach the membranous urethra. We present data to support port placement using the pubis, rather than the umbilicus, as the landmark. MATERIALS AND METHODS: If we assume the 25-cm working distance of the robot arm (Z) equals the hypotenuse of a triangle and the Y axis is the sum of the distance from the membranous urethra to the skin (Y1) plus the displacement of the skin secondary to CO(2) insufflation (Y2), then the horizontal distance X is from the robot port site to the pubis. To ascertain Y1, we randomly selected the CT scans of 25 men and measured the depth from the skin over the pubis to the membranous urethra. To determine Y2, we measured the change in height from the table of the port site after CO(2) insufflation in 11 robotic laparoscopic prostatectomies. RESULTS: The average distance of Y1 was 11 cm; Y2 was 6 cm. Using the formula (Z(2) - (Y1 + Y2)(2))(1/2), the maximum distance X from the port site to the pubis, for an average man, should not exceed 18 cm. CONCLUSION: The optimal landmark for calculating the placement of ports for the daVinci robotic arm placement should be the pubis and not the umbilicus. Tall men (>72 inches) are at risk for exceeding functional robot arm length, and in these men, port sites should not be more than 18 cm from the pubis. PMID- 15333226 TI - Laparoscopic radical prostatectomy. PMID- 15333227 TI - Use of bipolar energy for transurethral resection of bladder tumors: pathologic considerations. AB - BACKGROUND AND PURPOSE: Bipolar electrocautery has recently been introduced as a modality for transurethral resection of bladder tumors (TURBT). The primary benefits of bipolar TURBT stem from the use of saline irrigant rather than glycine or water. TURBT should be conducted in a fashion such that the resected tissue can be used for proper grading and staging, so excessive cauterization of the tissue should be avoided. In this study, we compared the pathologic characteristics of bladder tumor specimens resected with bipolar versus standard monopolar energy to determine specimen quality. PATIENTS AND METHODS: Bipolar TURBT (Gyrus Medical Inc., Maple Grove, MN) was performed in 11 patients. Pathologic specimens were compared with the specimens from 11 patients who had previously undergone standard monopolar TURBT. Resected tissue was examined by a pathologist who recorded tumor size, grade, location, presence of muscularis propria, presence of muscle invasion, and final diagnosis. The pathologist also determined the degree of cautery artifact in each specimen. The pathologist was blinded to the form of electrocautery used and the clinical diagnosis. RESULTS: Transurethral resection with bipolar electrocautery was carried out without difficulty or complication in all cases. Similarly, there were no complications in resection by standard monopolar electrocautery. The bladder tumor chips obtained with bipolar TURBT were smaller because of the smaller size of the bipolar loop. However, this did not interfere with the pathologic assessment. There were no significant pathologic differences between specimens according to the type of cautery used. A large degree of cautery artifact was noted in the tissue of larger tumors resected using both monopolar and bipolar electrocautery. However, the incidence and degree of cautery artifact were similar in the two groups. No trends between tumor location and degree of cautery effect were noted. The pathologist had no difficulty reaching a full and proper diagnosis in all cases involving either form of electrocautery. CONCLUSIONS: Bipolar electrocautery is well suited for TURBT. Bladder tissue obtained from bipolar TURBT is of the same histologic quality as that obtained from standard monopolar TURBT and provides the urologist with a reliable and complete diagnosis. PMID- 15333228 TI - Thermal artifacts in bladder tumors following loop endoresection: electrovaporization v electrocauterization. AB - BACKGROUND AND PURPOSE: Recently, the electrovaporization (EV) technique has been used for loop endoresection of bladder tumors. Our objective was to evaluate whether bladder tumors resected by EV with a loop wire are fit for histologic diagnosis. In addition, a quantitative comparison was made with the thermal artifacts created with a standard electrocautery (EC) loop resection. MATERIALS AND METHODS: In 26 patients with bladder tumors at various locations, endoresection was performed. In nonselective order, EV (N = 9; Gyrus device) or EC (N = 17; Valleylab device) was used. Histologic thermal artifacts were defined, and the diagnosis (pT classification) was determined. The linear depth of the thermal artifacts in the resected tissue was quantified with a computerized analysis system and statistically analyzed with the t-test. RESULTS: One case was excluded because the tissue was not fit for morphometric measurements. In all cases, a histologic diagnosis could be made. No qualitative differences were found between the groups in the extent of histologic thermal artifacts. The mean depth of the thermal artifact zone was 0.237 mm (range 0.060 0.469 mm; SD 0.098 mm) in the EV group and 0.260 mm (range 0.080-0.410 mm; SD 0.112 mm) in the EC group. This difference is not significant (P = 0.8). CONCLUSION: In these series, EV with a loop wire seemed to be a valid method to preserve bladder tumor specimens for histologic diagnosis. Compared with the EC method, there was no significant difference in the extent of thermal artifacts in the tissues resected. PMID- 15333229 TI - The effects of emotional salience, cognitive effort and meta-cognitive beliefs on a reality monitoring task in hallucination-prone subjects. AB - OBJECTIVES: A tendency to externalize internal information on reality monitoring tasks has been documented in psychiatric patients with hallucinations. Furthermore, previous studies suggest that factors such as the emotional salience of the material, cognitive effort and meta-cognitive beliefs are important contributory factors in this tendency to externalize internal information on reality monitoring tasks. However, few studies have investigated these aspects in hallucination-prone subjects. Also, these factors have never been examined simultaneously. In the following study we wished to examine the effects of emotional salience, cognitive effort and meta-cognitive beliefs on reality monitoring functioning in hallucination-prone subjects. DESIGN: Between participants group design. METHOD: One hundred normal subjects were administered a reality monitoring task. Words were presented by the experimenter. After each word, subjects were asked to say the first word that came to their mind. Words varied in terms of emotional valence and cognitive effort (high cognitive effort for words requiring longer latency times to associate a word and vice versa). Following a delay, words were presented consisting of those already presented by the experimenter or the subject (old) and those never presented before (new). For each word, subjects were required to identify whether the word was old or new. If the word was identified as old, subjects were required to identify the source of the word (subject or experimenter). Subjects also completed a questionnaire assessing meta-cognitive beliefs. RESULTS: Subjects were grouped according to their scores on a revised and elaborated version of the Launay-Slade Hallucinations Scale (LSHS). Those with scores within the top 25% were included in the hallucination-prone group (HP) (N = 25), whereas scores within the lower 25% were included in the non-hallucination-prone group (NHP) (N = 25). Results showed that the HP subjects had significantly more source discrimination errors than NHP subjects for self-generated items. In other words, HP subjects tended to misattribute to the experimenter items that they had produced themselves. This pattern was especially marked with emotionally charged material and with words that required more cognitive effort. In addition, HP subjects scored significantly higher on a scale assessing meta-cognitive beliefs compared with NHP subjects. Finally, scores on a scale assessing meta-cognitive beliefs were positively associated with source discrimination errors. CONCLUSIONS: These results suggest that cognitive effort, emotional salience and meta-cognitive beliefs all play a prominent role in the externalizing bias in hallucination prone subjects. The results also provide evidence for the validity of the idea of a continuity between hallucination-prone subjects and psychotic patients with hallucinations on reality monitoring tasks, including a number of contributing factors in the occurrence of hallucinations. PMID- 15333230 TI - Viewing behaviour of spider phobics and non-phobics in the presence of threat and safety stimuli. AB - OBJECTIVES: To determine whether spider phobics and spider non-phobics differ in attending to threat and safety stimuli in close proximity, or spatially separated. METHOD: A sample group of 16 spider-phobic and 16 spider-non-phobic volunteers were drawn from a student population on the basis of their scores on Watts and Sharrock's Spider Phobia Questionnaire. Attention was assessed by way of participants' viewing behaviour using an ASL 501 head-mounted eye-tracking system. In a control condition, participants viewed a neutral stimulus (a TV video) in the absence of the threat stimulus. In two experimental conditions, the threat stimulus (a live Chilean rose tarantula) was introduced (a) immediately beside the safety stimulus (the only exit of the experimental room), or (b) away from the safety stimulus. RESULTS: In the experimental conditions, phobics reduced their viewing of the TV and increased their viewing of the tarantula and of the exit; the biggest changes occurred when tarantula and exit were together. Phobics also made more eye motions across the experimental room. CONCLUSION: Our results are consistent with previous research employing reaction time to a neutral stimulus as dependent measure. In addition, our results suggest that phobics scan the environment as part of safety behaviour. We suggest that exposure treatments to reduce spider phobia may be facilitated by encouraging patients to stop environmental scanning. PMID- 15333231 TI - The positive and negative affect schedule (PANAS): construct validity, measurement properties and normative data in a large non-clinical sample. AB - OBJECTIVES: To evaluate the reliability and validity of the PANAS (Watson, Clark, & Tellegen, 1988b) and provide normative data. DESIGN: Cross-sectional and correlational. METHOD: The PANAS was administered to a non-clinical sample, broadly representative of the general adult UK population (N = 1,003). Competing models of the latent structure of the PANAS were evaluated using confirmatory factor analysis. Regression and correlational analysis were used to determine the influence of demographic variables on PANAS scores as well as the relationship between the PANAS with measures of depression and anxiety (the HADS and the DASS). RESULTS: The best-fitting model (robust comparative fit index = .94) of the latent structure of the PANAS consisted of two correlated factors corresponding to the PA and NA scales, and permitted correlated error between items drawn from the same mood subcategories (Zevon & Tellegen, 1982). Demographic variables had only very modest influences on PANAS scores and the PANAS exhibited measurement invariance across demographic subgroups. The reliability of the PANAS was high, and the pattern of relationships between the PANAS and the DASS and HADS were consistent with tripartite theory. CONCLUSION: The PANAS is a reliable and valid measure of the constructs it was intended to assess, although the hypothesis of complete independence between PA and NA must be rejected. The utility of this measure is enhanced by the provision of large scale normative data. PMID- 15333232 TI - Causal attributions in delusional thinking: an investigation using qualitative methods. AB - OBJECTIVES: To investigate the attributions offered spontaneously during interview by patients who have delusional beliefs. Specifically, to assess whether people with delusional beliefs reason in a way that is analysable. If so, to extend the findings from previous research to provide a detailed picture of the patients' accounts that could inform cognitive therapies for psychosis. METHOD: A group of 31 patients with delusional beliefs were interviewed using a semi-structured open-ended interview. Their causal attributions were identified and analysed in terms of standard definitions of attributional theory using the Leeds Attributional Coding System. RESULTS: Patients made an average of 96 attributions each. These showed a high degree of recognizable coherence in reasoning, including when directly discussing delusional beliefs. The patterns of attributing were partly consistent with previous research findings, with external attributions being offered for negative events. Several other aspects emerged, indicating a pervasive negativity, with non-delusional aspects of the illness being a major concern. When delusions were described as a cause of events, the dominant pattern of attributing was stable, internal, personal and uncontrollable, with negative outcomes. When delusions were reported as an outcome, the patient was more in control. CONCLUSIONS: The reasoning processes of patients with delusional beliefs are in many aspects normal and include patterns not identified by the ASQ. Attributing in relation to delusions differs from talk about other subjects. The qualitative attributional approach provided a detailed and rich picture of the world of these patients. Interpretable tendencies are described that should provide information useful to those practising and evaluating cognitive therapies. PMID- 15333233 TI - A personality-based taxonomy of sexually abusive adolescents derived from the Millon Adolescent Clinical Inventory (MACI). AB - A personality-based taxonomy of an out-patient sample of 112 sexually abusive adolescents is described. Five subgroups were derived from cluster analytic procedures applied to Personality Patterns scales scores from the Millon Adolescent Clinical Inventory (MACI), and were named: Normal, Antisocial, Submissive, Dysthymic/Inhibited, and Dysthymic/Negativistic. The groups were also found to be differentiated on the psychopathology scales of the MACI. The results of the study provide evidence of the heterogeneity of adolescent sexual abusers in both personality characteristics and psychopathology. Comparison of groups differentiated on the basis of victim characteristics did not indicate a relationship between personality and sexual offence. PMID- 15333234 TI - The role of body objectification in disordered eating and depressed mood. AB - OBJECTIVES: The study aimed to extend tests of objectification theory into the realm of depression. The theory's applicability to men was also investigated. DESIGN: A cross-sectional study. METHOD: A sample of 115 men and 171 women completed questionnaire measures of self-objectification, depressed mood, disordered eating, as well as the proposed mediating variables of body shame, appearance anxiety, flow and awareness of internal states. RESULTS: For women, it was found that depressed mood and disordered eating were both predicted by self objectification and its corollary of habitual self-surveillance. Path analysis gave strong support to the mediational relationships of the theoretical model. With one major exception (the role of self-objectification), the pattern of relationships was similar for men. CONCLUSION: Objectification theory provides a useful framework for identifying predictors of depressed mood. PMID- 15333235 TI - The use of a cognitive battery to predict who will fail an on-road driving test. AB - OBJECTIVES: There is a growing need to find a valid and reliable neuropsychological battery to screen out those people who are clearly unsafe to drive following brain injury or pathology, and thus alleviate the need to refer for an on-road assessment. DESIGN: A battery of cognitive tests fine-tuned for their relevance to driving was examined in terms of its sensitivity and specificity for predicting who would fail an on-road test following brain injury or pathology. METHOD: Performance on the battery was compared to the results of an on-road driving test in a consecutive series of 142 clients referred to a driving assessment centre following brain injury or pathology. The group represented diverse neurological conditions which affect brain functioning. RESULTS: The overall accuracy rate of the battery in predicting a fail on-road was 92% and in predicting a pass on road was 71%. It was more accurate for those under 70 with 100% accuracy in predicting a fail on-road and 85% accuracy in predicting a pass on-road, but less accurate for those aged 70 or above with 85% accuracy in predicting a fail on-road and 37% accuracy in predicting a pass on road. CONCLUSION: The battery is a useful tool in helping to determine whether someone is safe to drive following brain injury but needs to be used with far more caution for the older driver. PMID- 15333236 TI - Cognitive representations in alcohol and opiate abuse: the role of core beliefs. AB - There are clinical indications that alcohol and drug abuse are associated with unhealthy core beliefs (unconditional, schema-level representations). This study examined levels of such cognitions among four groups: alcohol abusers; opiate abusers; combined alcohol and opiate abusers; and a non-clinical group. Each patient completed the short version of the Young Schema Questionnaire, measuring levels of 15 pathological core beliefs. These schema-level cognitions were less healthy in the clinical groups than in the non-clinical group, particularly among individuals who abused alcohol. These findings provide preliminary support for the utility of therapies that address schema-level representations among substance abusers. PMID- 15333237 TI - The new Italian IVF legislation. AB - Last February, the Italian Parliament gave final approval to a new Law regulating assisted reproduction technology. The new legislation fell short of the expectations of infertile couples and of all specialists in the field. There are three problems with the new Italian law; they involve social issues, human rights and the application of technology. The present paper focuses on the fact that the new rules infringe upon basic human rights and the proper application of IVF technology, because they mandate procedures that are against the best interest of the woman seeking pregnancy. The main point of controversy is the combination of a mandatory limit of three embryos for transfer, and an obligation to reimplant all produced embryos; cryopreservation of excess embryos is prohibited. Obviously, this decreases the chances of most women to achieve pregnancy, while at the same time it increases the number and complexity of procedures they need to undergo and may expose some to an unacceptable increase in the risk of multiple pregnancy. The new law is inspired by the desire to protect every newly produced embryo; this is a commendable aim, although it is in total opposition to a law passed over 25 years ago that liberalized voluntary termination of first trimester pregnancies. This means that today Italy has a law that protects every early, pre-implantation embryo, and another that allows the 'suppression' of every post-implantation one. From a technical point of view, given the low level of human fecundity, the only way to prevent the 'loss' of even one preimplantation embryo is to simply ban IVF altogether, an option that Italian legislators obviously did not have the courage to opt for. The tragedy is that Italian infertile couples are now confronted with new rules that not only severely limit the ability of physicians to correctly apply IVF technology, but are so confused that, depending on the interpretation, anyone may try to nullify the main ideological premise upon which the entire law has been structured. PMID- 15333239 TI - Concerns regarding the follow-up of children conceived after assisted reproduction in Latin America. AB - Children born through assisted reproduction are not usually followed up in Latin America. However, in spite of all the social-economic and cultural complexity of this part of the world, professionals involved in assisted reproduction should not relinquish the responsibility of following up these children. In May 2002, the Centre for Human Reproduction Sinha Junqueira, started a specific project for evaluation and follow-up of its children. It is the first centre in Brazil, and probably in Latin America, with this aim, and this paper reports initial results concerning the intellectual development of these children. PMID- 15333240 TI - Further thoughts regarding evidence offered in support of the 'Barker hypothesis'. AB - The original objection to the paper by Kwong et al. was that the use of an inappropriate (between-pup) estimate of experimental error had exaggerated the importance of the maternal nutrition effect. From the group's most recent response, it has been possible to regenerate the raw data and carry out a further detailed analysis. It is apparent that despite now using a more sophisticated statistical tool, Kwong et al. have still, in effect, used the between-pup error, thus repeating the previous, probably exaggerated, finding. It is maintained that the nutrition effect should be studied using the between-rat variation, which then provides a result that is a good deal less emphatic. Further, it is felt that there is a very important point of principle involved in this dispute, relating to the rigorous analysis of hierarchical data, particularly in small studies. PMID- 15333241 TI - Cryopreservation and assisted human conception. Introduction. PMID- 15333242 TI - Current and future concepts and practices in human sperm cryobanking. AB - This article focuses on a range of issues that are of current concern to those working in human sperm cryobanking and considers them within a general framework of risk analysis and management, taking into account the available evidence and perceptions of 'best practice' from both the medical and legal perspectives. In particular, issues arising from concerns over the risk of cross-contamination between samples during storage immersed in liquid nitrogen have been analysed in detail. Even though such an occurrence has never been reported for spermatozoa or embryos, and the risk is generally accepted to be vanishingly small, it does represent a finite risk and all reasonable measures should be taken to reduce the chance of its occurring. It is concluded that all methods used to collect, cryopreserve, store, thaw and use cryobanked human spermatozoa must address the risk of contamination from any source throughout the entire process. To assist workers in this area, a series of recommendations as to current best practice are made, based upon conclusions from risk analyses using currently available information. PMID- 15333244 TI - An overview of oocyte cryopreservation. AB - The ability to cryopreserve human oocytes and store them indefinitely would be beneficial for cancer patients at risk of becoming sterile after therapy, allow women to delay reproduction, and alleviate religious concerns associated with embryo storage. In 1986, Chen was the first to report a pregnancy originating from a frozen-thawed human oocyte. Although over 100 babies have been born from oocyte storage since then, pregnancy rates remain unacceptably low. Adapting embryo cryopreservation techniques to oocyte storage has had limited success and new reproducible methods are needed. Problem areas other than intracellular ice formation and osmotic effects need to be identified. A broad approach of critical analysis should be conducted regarding the entire cryopreservation process from pre-equilibration and cooling, to thawing and stepout. All established facets deserve reanalysis in order to assess which aspects can be optimized or changed so that cellular demise can be avoided and cellular viability enhanced. New methods, including the use of choline-based media and vitrification have proven useful in increasing survival and pregnancy rates in some clinics. Other methods yet untested, such as injection of complex carbohydrates into the oocyte, deserve further studies. Vitrification research has led to the formulation of new ideas and has demonstrated the flexibility of cells to survive cryopreservation. Although successful, vitrification protocols are potentially harmful and technically challenging, due to elevated cryoprotectant concentrations and rapid cooling rates. Bovine embryo vitrification methods have been used to store human oocytes and embryos, particularly blastocysts with some success. Vitrification solutions containing high molecular weight polymers have also proved beneficial by reducing solution toxicity. In general, further advances are needed to improve human oocyte storage before widespread routine clinical use. PMID- 15333245 TI - Cryopreservation of animal and human embryos by vitrification. AB - Vitrification is a method in which not only cells but also the whole solution is solidified without the crystallization of ice. For embryo cryopreservation, the vitrification method has advantages over the slow freezing method. For example, injuries related to ice is less likely to occur, embryo survival is more likely if the embryo treatment is optimized, and embryos can be cryopreserved by a simple method in a short period without a programmed freezer. However, solutions for vitrification must include a high concentration of permeating cryoprotectants, which may cause injury through the toxicity of the agents. Since the development of the first vitrification solution, which contained dimethylsulphoxide, acetamide, and propylene glycol, numerous solutions have been composed and reported to be effective. However, ethylene glycol is now most widely used as the permeating component. As supplements, a macromolecule and/or a small saccharide are frequently added. Embryos of various species, including humans, can be cryopreserved by conventional vitrification using insemination straws or by ultrarapid vitrification using minute tools such as electron microscopic grids, thin capillaries, minute loops, or minute sticks, or as microdrops. In the ultrarapid method, solutions with a lower concentration of permeating cryoprotectants, thus having a lower toxicity, can be used, because ultrarapid cooling/warming helps to prevent ice formation. PMID- 15333247 TI - Developmental consequences of cryopreservation of mammalian oocytes and embryos. AB - During the last three decades, significant advances have been made in successful cryopreservation of mammalian preimplantation embryos, and more recently oocytes. The ability to cryopreserve, thaw, and establish pregnancies with supernumerary preimplantation embryos has become an important tool in fertility treatment. Human oocyte cryopreservation has practical application in preserving fertility for individuals at risk of compromised egg quality due to cancer treatments or advanced maternal age. While oocyte/embryo cryopreservation success has increased over time, there is still room for improvement. Oocytes and embryos are susceptible to cryo-damage, which collectively entails cellular damage caused by mechanical, chemical, or thermal forces during the freeze-thaw process. Basic studies focused on understanding cellular structures, their composition, and more importantly their functions, in normal cell developments will continue to be critical in assessing, understanding, and correcting oocyte/embryo cryo-damage. This review will delineate many of the oocyte/embryo intracellular and extracellular structures that are or may be compromised during cryopreservation. A global theme presented throughout this review is that many structural components of the oocyte/embryo also have essential functional roles in development. Compromising these cellular structures, and thus their cellular homeostatic functions, can deleteriously influence initial cryo-survival or compromise subsequent normal development through effects on the oocyte and/or early embryo. PMID- 15333248 TI - Use of sugars in cryopreserving human oocytes. AB - In the last 20 years, a worldwide effort to cryopreserve oocytes has resulted in 40 infants and approximately 50 ongoing pregnancies being reported. While the ability to freeze human embryos has become a standard of practice in assisted reproductive technologies, obtaining reliable techniques for oocyte cryopreservation has been more difficult. The unique properties of the mature oocyte, such as the meiotic stage with sensitive spindle structure as well as the large cell volume, are responsible for the limited success obtained to date. There have been two approaches to cryopreserving the oocyte: (i) slow freeze rapid thaw, and (ii) vitrification protocols with rapid cooling-rapid warming. Both methods have incorporated sugars (sucrose) as a beneficial non-permeating extracellular cryoprotectant. Studies of organisms that survive extreme conditions of freezing/dehydration have demonstrated the ability to accumulate intracellular sugars to afford protection and survival. A novel technique using microinjection of sugars into the oocyte for cryopreservation has been developed as an alternative approach to external addition of sugars. Freezing the human oocyte has been a challenging goal; however, developing research and efforts will, in the near future, provide women with an important option for their reproductive health. PMID- 15333249 TI - Comparison of necrosis in human ovarian tissue after conventional slow freezing or vitrification and transplantation in ovariectomized SCID mice. AB - This paper examines and compares necrosis in human ovarian tissue after conventional slow freezing or vitrification and ensuing xenotranplantation. Slow cryoconserved or vitrified ovarian tissue samples and fresh controls from nine patients were subcutaneously transplanted into SCID mice. The tissue samples were explanted after 6 weeks and the necrotic areas were examined by staining with Lucifer yellow SV. The size of the necrotic areas in parallel cultivated ovarian tissue samples was compared, as was necrosis in cultivated prostate tumour spheroids where the emergence of necrosis and its pathophysiological correlation have been described. Examinations showed no significant rise in the proportion of necrotic areas after slow cryoconservation/transplantation and in the controls (transplanted fresh tissue, not transplanted fresh tissue, long-term culture). The proportion of necrotic areas in the tumour spheroids was significantly higher than in the ovarian tissue. Vitrification could, after these results, be presented as an alternative to conventional slow cryoconservation. PMID- 15333250 TI - CYP1A1, GSTM1 and GSTT1 genetic polymorphism is associated with susceptibility to polycystic ovaries in South Indian women. AB - Polymorphic variants in the phase I enzyme, cytochrome P450 gene, may lead to increased toxification, whereas polymorphisms in the phase II enzyme, glutathione S-transferase genes, may result in impaired detoxification. Alterations in the activities of phase I drug metabolizing enzymes and phase II detoxification enzymes may cause abnormal functioning and formation of follicular cysts in the ovaries and thus causing an imbalance in the hormone profiles. This study aimed to investigate the relationship between genetic polymorphisms of CYP1A1 (T6235C), GSTM1 and GSTT1 in South Indian women with polycystic ovaries (PCO) using polymerase chain reaction-restriction fragment length polymorphism. The frequencies of variants of these genes were studied in 180 women with confirmed PCO and in 72 healthy fertile women with successful pregnancy record. No significant difference was found between the frequencies of GSTM1 and GSTT1 null genotypes in PCO cases and healthy controls. However, CYP1A1 Msp I homozygous mutants were strongly associated (P = 0.0139) with increased susceptibility to PCO. Three genotype combinations, CYP1A1 (mt/mt) with GSTM1 [-] and GSTT1 [-], CYP1A1 (wt/mt) with GSTM1 [-] and GSTT1 [-] and CYP1A1 (mt/mt) with GSTM1 [-], GSTT1 [+], were also observed in women with PCO. In conclusion, the presence of hyperinducible CYP1A1 (T6235C) mutant genotype and its mutants in combination with GSTM1 and GSTT1 null genotypes might cause an imbalance between phase I and phase II enzymes, and therefore may represent a risk factor for PCO. PMID- 15333252 TI - Risk factors for the deterioration of glucose metabolism in polycystic ovary syndrome. AB - Women with polycystic ovary syndrome (PCOS) have a high risk of abnormal glucose metabolism, which deteriorates gradually over time. This study was designed to assess contributing factors to the rate of deterioration of glucose metabolism over the 4- to 7-year follow-up period. The study group included 67 PCOS women who had normal glucose tolerance in the initial test in 1990-1992. There was a significant increase in both fasting and 2-h glucose concentrations, and body weight, over the follow-up period. The initial obesity, expressed as waist:hip ratio and BMI, was significantly related to the increase of glucose concentrations, while weight gain was not a significant factor. The overweight/obese PCOS women have a faster deterioration of glucose metabolism over the follow-up period than the non-obese ones. Moderate weight gain has no significant detrimental effect on the rate of deterioration of glucose metabolism. PMID- 15333254 TI - Preimplantation genetic diagnosis with HLA matching. AB - Preimplantation genetic diagnosis (PGD) has recently been offered in combination with HLA typing, which allowed a successful haematopoietic reconstitution in affected siblings with Fanconi anaemia by transplantation of stem cells obtained from the HLA-matched offspring resulting from PGD. This study presents the results of the first PGD practical experience performed in a group of couples at risk for producing children with genetic disorders. These parents also requested preimplantation HLA typing for treating the affected children in the family, who required HLA-matched stem cell transplantation. Using a standard IVF procedure, oocytes or embryos were tested for causative gene mutations simultaneously with HLA alleles, selecting and transferring only those unaffected embryos, which were HLA matched to the affected siblings. The procedure was performed for patients with children affected by Fanconi anaemia (FANC) A and C, different thalassaemia mutations, Wiscott-Aldrich syndrome, X-linked adrenoleukodystrophy, X-linked hyperimmunoglobulin M syndrome and X-linked hypohidrotic ectodermal displasia with immune deficiency. Overall, 46 PGD cycles were performed for 26 couples, resulting in selection and transfer of 50 unaffected HLA-matched embryos in 33 cycles, yielding six HLA-matched clinical pregnancies and the birth of five unaffected HLA-matched children. Despite the controversy of PGD use for HLA typing, the data demonstrate the usefulness of this approach for at-risk couples, not only to avoid the birth of affected children with an inherited disease, but also for having unaffected children who may also be potential HLA-matched donors of stem cells for treatment of affected siblings. PMID- 15333255 TI - Ethics of PGD: thoughts on the consequences of typing HLA in embryos. AB - As with so many fields of study associated with assisted human reproduction, many ethical issues are raised by the practice of preimplantation diagnosis of inherited disease (PGD). Some are part and parcel of assisted conception, e.g.the rights of human embryos in vitro and of embryologists to establish them, carry out research and discard them. Others unique to clinical PGD were discussed at an earlier meeting on PGD (Edwards et al., 2003). Recent developments in PGD are discussed briefly in this Commentary, especially the ethics of designer babies. PMID- 15333256 TI - Paternal gonadal mosaicism detected in a couple with recurrent abortions undergoing PGD: FISH analysis of sperm nuclei proves valuable. AB - Many couples are now seeking preimplantation genetic diagnosis (PGD) and fluorescence in-situ hybridization (FISH) as an alternative approach to avoid spontaneous abortion by ensuring transfer of presumed chromosomally normal embryos. This case report describes unexpected findings in a couple having three spontaneous abortions and two failed IVF cycles. In two IVF PGD cycles, four of 13 (30.8%) embryos (blastomeres) demonstrated duplication involving the Down syndrome critical region, detectable by a locus specific chromosome 21 probe. The same duplication was subsequently detected by FISH in 66 of 1002 (6.6%) sperm nuclei, demonstrating paternal gonadal mosaicism. Cytogenetic studies of peripheral blood revealed normal karyotypes in both the male and female partners. This identification of paternal germ cell or gonadal mosaicism suggests that analysis of sperm nuclei prior to undergoing IVF with PGD may be of value in patients with recurrent spontaneous abortions or multiple failed IVF. PMID- 15333258 TI - FSH isoform composition of commercial gonadotrophin preparations: a neglected aspect? AB - The clinical efficacy of commercial gonadotrophin preparations has been the subject of an intense debate during recent years. Arguments have primarily focused on the origin of FSH activity (urine versus recombinant derived) and whether the preparation included LH-like activity. FSH isoform composition has received little or no attention, and is usually considered to have negligible effect on clinical effectiveness. By presenting the available data on the FSH isoform composition of commercial gonadotrophin preparations, the present paper challenges this assumption. To evaluate whether the FSH isoform composition affected the efficacy of a product, a meta-analysis was performed that compared a preparation expressing an acidic isoform profile (urinary-derived Metrodin-HP) with a preparation rich in less acidic isoforms (recombinant derived Gonal F). A total of five randomized clinical trials that specifically compared these two preparations was identified and included in the analysis. All parameters relating to the direct effect of FSH on the follicle differed significantly in favour of the product rich in less acidic isoforms, while data on pregnancy outcome did not reach significance. The importance of the FSH isoform profile and whether the FSH is derived from urine or by recombinant technique is discussed in relation to clinical efficacy. It is suggested that the FSH isoform profile of commercial gonadotrophin preparations is of clinical importance and should be taken into account when evaluating efficacy. PMID- 15333259 TI - Why and how should multiple pregnancies be prevented in assisted reproduction treatment programmes? AB - Although most professional societies have issued guidelines to diminish the number of embryos to be transferred during assisted reproductive techniques, the incidence of multiple pregnancies remains unacceptably high. The burden of morbidity and mortality seems to increase substantially with each fetus in a multiple gestation. As a result, there has been growing debate on the need to prevent multiple pregnancies. The infertility specialists who can solve the infertility problem are usually shielded from the complications of multiple pregnancies. If they were involved in the delivery and, more particularly in the care of multiple pregnancies (both financially and socially), their attitude would probably change. IVF centres should gradually reduce the mean number of embryos per transfer in terms of the cost:benefit ratio. A further reduction to one single embryo per transfer in good cases would be similarly acceptable. Laboratory expertise is of vital importance, especially in terms of embryo culture, embryo selection, and freezing and thawing techniques in embryo transfer programmes for reducing the number of transferred embryos. PMID- 15333260 TI - Risk factors for childbearing during adolescence in a population-based birth cohort in southern Brazil. AB - OBJECTIVE: To use a case-control study to analyze risk factors associated with teenage childbearing among adolescents who were in a birth cohort study that began in 1982 in the city of Pelotas, Rio Grande do Sul, Brazil. METHODS: Adolescent mothers in Pelotas who gave birth between January 1995 and March 2001 and who had been born in 1982 were identified through the local birth information system. These subjects from the 1982 birth cohort were compared to adolescents from the same cohort who had not given birth before March 2001. Standardized interviews were used in 2001 to obtain information about socioeconomic, maternal reproductive, demographic, and lifestyle characteristics. This information was combined with data obtained in earlier phases of this 1982 birth cohort study. Unconditional logistic regression was used to analyze the risk factors associated with childbearing during adolescence. RESULTS: A total of 420 parous adolescents from the 1982 birth cohort were identified and then compared with 408 cohort adolescents who had not given birth by March 2001. Higher family income in 1982 and more parental schooling in 1982 were inversely related to childbearing among the birth cohort adolescents. Cohort girls whose mothers were under age 20 when they gave birth in 1982 had a higher risk of becoming pregnant while still an adolescent. Cohort girls who, during childhood, lived with siblings from different fathers were twice as likely to become an adolescent mother. Compared to cohort adolescents who had not failed during the first four years of school, those girls who had done so had twice the risk of giving birth during adolescence. Among the cohort girls a positive association was found between younger age at first intercourse and childbearing in adolescence. CONCLUSIONS: The results indicate that the educational level of the adolescent mothers must be considered in planning policies that attempt to disrupt successive cycles of socioeconomic deprivation. Early interventions to improve sex education and to increase the motivation of young girls to achieve higher levels of education are needed. The interventions should aim for a reduction in the risk of long-term poverty and poorer educational achievements of teenage mothers and their children. PMID- 15333261 TI - [Tiredness and sleepiness in bus drivers and road accidents in Peru: a quantitative study]. AB - OBJECTIVE: To evaluate the relationship that tiredness and sleepiness in bus drivers have to road accidents in Peru. Information from various countries indicates that driver sleepiness plays an important role in road accidents. However, there is only limited information on this subject in Peru. METHODS: Using a supervised, pretested survey, a cross-sectional observational and comparative study was carried out with 238 bus drivers who drive on the Northern Pan American Highway of Peru. To determine the relationship between variables the chi-square test was used, along with the Pearson correlation coefficient. The level of significance was set at P < 0.05. The variables analyzed were: tiredness, sleepiness, hours of driving per day, daily hours of sleep, body mass index, snoring, sleep apnea, and either having had or almost having had an accident while driving. RESULTS: Of the 238 drivers, all of them were men, 45% said they had had or nearly had had an accident while driving, 55% slept less than 6 hours per day, 31% had slept less than 6 hours in the 24 hours before answering the survey, and 80% were in the habit of driving more than 5 hours without stopping. Of the drivers, 56% of them reported being tired at least some of the time while driving; of this group, 65% of them reported being tired during the early morning. Seventy-six drivers (32%) said that while they were driving their eyes had fallen shut. In terms of where they slept, 194 of the drivers (81%) said they always slept in the lower luggage compartment of the bus while another driver was driving the bus or when the bus was parked in the bus terminal. The steps that drivers took to avoid falling asleep while driving included: wetting the face with water, eating fruit, opening the window of the driver's compartment, drinking coffee, listening to music, smoking, chewing coca leaves, and drinking alcohol mixed with coca leaves. In the opinion of 55% of the drivers, the leading cause of road accidents is tiredness. Accidents and near accidents while driving occurred mainly between midnight and 6 a. m. Having an accident or a near-accident was strongly associated with tiredness and with having the eyes drop shut while driving (P < 0.0005). CONCLUSIONS: Tiredness and sleepiness while driving were common among the bus drivers, with various possible causes: acute and chronic sleep deprivation, irregular schedule changes, and sleep disorders due to the drivers' working conditions. Our results support the hypothesis that fatigue and sleepiness among bus drivers are related to road accidents. PMID- 15333262 TI - [Foremilk and hindmilk retinol levels]. AB - OBJECTIVE: To determine retinol levels in breast milk at the beginning and at the end of a feeding. METHODS: In 2003, 30 breastfeeding women receiving care at the Januario Cicco Maternity School, in the city of Natal, Rio Grande do Norte, Brazil, were interviewed between 24 hours and 15 days after delivery. A four mL sample of breast milk was expressed manually (2 mL at the beginning and 2 mL at the end of the feeding) in the afternoon, two hours after the preceding feeding. Retinol levels were determined by high performance liquid chromatography. Student's t test was used for statistical analysis. RESULTS: The mean retinol concentrations in the breast milk samples collected at the beginning and at the end of the same feeding were 69.3 +/- 41.4 microg/100 mL and 111.6 +/- 79.2 microg/100 mL, respectively; the mean concentrations were statistically different (P = 0.012). CONCLUSION: Our results suggest that hindmilk provides a higher intake of vitamin A. Therefore, it is important to guide mothers to not limit the baby's time at the breast and to avoid expressing only foremilk. In addition, it is important to standardize the time of breast milk collection in order to allow comparisons between the results of different studies on this topic. PMID- 15333263 TI - Tuberculosis along the United States-Mexico border, 1993-2001. AB - OBJECTIVES: Tuberculosis (TB) is a leading public health problem and a recognized priority for the federal Governments of both Mexico and the United States of America. The objectives of this research, primarily for the four states in the United States that are along the border with Mexico, were to: (1) describe the epidemiological situation of TB, (2) identify TB risk factors, and (3) discuss tuberculosis program strategies. METHODS: We analyzed tuberculosis case reports collected from 1993 through 2001 by the tuberculosis surveillance system of the United States. We used those data to compare TB cases mainly among three groups: (1) Mexican-born persons in the four United States border states (Arizona, California, New Mexico, and Texas), (2) persons in those four border states who had been born in the United States, and (3) Mexican-born persons in the 46 other states of the United States, which do not border Mexico. RESULTS: For the period from 1993 through 2001, of the 16 223 TB cases reported for Mexican-born persons in the United States, 12 450 of them (76.7%) were reported by Arizona, California, New Mexico, and Texas. In those four border states overall in 2001, tuberculosis case rates for Mexican-born persons were 5.0 times as high as the rates for persons born in the United States; those four states have 23 counties that directly border on Mexico, and the ratio in those counties was 5.8. HIV seropositivity, drug and alcohol use, unemployment, and incarceration were significantly less likely to be reported in Mexican-born TB patients from the four border states and the nonborder states than in patients born in the United States from the four border states (P < 0.001). Multivariate analysis revealed that among pulmonary tuberculosis patients who were 18-64 years of age and residing in the four border states, the Mexican-born patients were 3.6 times as likely as the United States-born patients were to have resistance to at least isoniazid and rifampin (i. e., to have multidrug-resistant TB) and twice as likely to have isoniazid resistance. Mexican-born TB patients from the four border states and the nonborder states were significantly more likely to have moved or to be lost to follow-up than were the TB patients born in the United States from the four border states (P < 0.001). CONCLUSIONS: Increased collaborative tuberculosis control efforts by the federal Governments of both Mexico and the United States along the border that they share are needed if tuberculosis is to be eliminated in the United States. PMID- 15333264 TI - Impaired glucose regulation in adults in Jamaica: who should have the oral glucose tolerance test? AB - OBJECTIVE: To compare the 1999 World Health Organization (WHO) fasting plasma glucose (FPG) criteria and the WHO 2-hour post-challenge glucose (2hPG) criteria during an oral glucose tolerance test (OGTT) in identifying adults in Jamaica with hyperglycemia. As the OGTT is not commonly used in clinical practice, factors associated with the failure of the FPG criteria to detect persons with impaired 2hPG were investigated. METHODS: A random sample of 2 096 adults, 25-74 years old, living in the town of Spanish Town, Jamaica, was evaluated for diabetes. After excluding 215 individuals for reasons such as missing data, the remaining 1 881 persons were composed of 187 who were previously known to have diabetes and 1 694 who were screened for diabetes with both FPG and 2hPG. RESULTS: The FPG criteria detected 83 cases of diabetes, compared to 72 by the 2hPG criteria. The kappa statistic comparing the two criteria was 0.31 (95% confidence interval: 0.28-0.34), indicating fair agreement. There were 261 cases of impaired glucose tolerance (IGT) and 92 cases of impaired fasting glucose (IFG). In those 92 with IFG, an OGTT would identify 34 cases of IGT and 14 cases of diabetes. Of those classified as normoglycemic by FPG criteria, 14% of them had IGT or diabetes by 2hPG criteria. The factors predicting the likelihood of non-detection of impaired glucose tolerance or diabetes by FPG were age, body mass index, central obesity, systolic blood pressure, and female sex. By receiver operating characteristic curve analysis, an FPG of 5.1 mmol/L would predict a 2hPG >/= 7.8 mmol/L. CONCLUSIONS: A few individuals classified as normal on FPG will have IGT or diabetes, and an OGTT will be needed to identify them. The yield of IGT detected by screening in Jamaica can be improved by lowering the threshold for IFG or by using clinical information to identify high-risk individuals. PMID- 15333265 TI - [Violence and social representation in teenagers in Brazil]. AB - OBJECTIVE: To investigate the association between the self-representation of teenagers and the severe physical, psychological, and sexual violence inflicted on them by close family relations, especially their parents, and to analyze the association between victimization in the family and victimization in other social spaces. METHOD: An epidemiological survey was carried out in 2002 with 1 685 adolescents chosen at random from public and private schools in the municipality of Sao Goncalo, which is in the state of Rio de Janeiro, Brazil. To measure each form of violence, we used scales of tactics for dealing with conflict, of child abuse and trauma, and of psychological violence. RESULTS: We found that 14.6% of the students had been physically abused by the father or the mother and that 11.8% had witnessed sexual abuse of another family member or they themselves had been sexually abused. In addition, 48.0% of the students reported having been psychologically abused by a close relation. In comparison to students who had not been abused, the victims of family abuse were more often also victims of community and school violence, and they also more frequently reported having broken the law. Overall, the adolescents surveyed had a positive self representation, but the adolescents who had been abused mentioned negative self attributes more frequently than did the teenagers who had not been abused. CONCLUSIONS: The predominantly positive social representation of teenagers must be supported by health promotion initiatives. The finding of an association between indices of violence and the teenager's various spheres of action indicates that resolving this problem will require strategies that target all these spheres. PMID- 15333266 TI - [Editorial policies approved by the CSE Board of Directors]. PMID- 15333267 TI - [Multiple drug resistance: a threat for tuberculosis control]. AB - Drug-resistant tuberculosis (TB) was reported soon after the introduction of streptomycin, although it did not receive major attention until recently. It was not considered a major issue in the industrialized world until outbreaks of multidrug-resistant TB (MDR-TB) were reported among HIV infected people. Administration of standard short-course chemotherapy (SSCC) with first-line drugs under directly observed therapy (DOT) is the cornerstone of modern TB control. Unfortunately, data available on the treatment outcome of MDR-TB cases under routine programmatic conditions suggest that patients with MDR-TB respond poorly to SSCC with first-line drugs. Since 1994, the World Health Organization and the International Union Against Tuberculosis and Lung Disease (IU-ATLD) have conducted anti-TB drug resistance surveys through a network of subregional laboratories and researchers. Drug resistance was present in almost all settings surveyed, and prevalence varied widely across regions. High prevalence of MDR-TB is widespread in the Russian Federation and areas of the former Soviet Union (Estonia, Kazakhstan, Latvia, and Lithuania) as well as Israel, Liaoning and Henan Provinces in China, and Ecuador. The Global Project has surveyed areas representing over one third of notified TB cases. However, enormous gaps still exist in the most crucial areas. The most effective strategy to prevent the emergence of drug resistance is through implementation of the directly observed treatment short (DOTS) strategy. Effective implementation of the DOTS strategy saves lives through decreased TB transmission, decreased risk of emergence of drug-resistance, and decreased risk for individual TB patients of treatment failure, TB relapse, and death. The World Bank recognizes the DOTS strategy as one of the most cost-effective health interventions, and recommends that effective TB treatment be a part of the essential clinical services package available in primary health care settings. Governments are responsible for ensuring the provision of effective TB control through the DOTS strategy. WHO and its international partners have formed the DOTS-Plus Working Group, which is attempting to determine the best possible strategy to manage MDR-TB. One of the goals of DOTS-Plus is to increase access to expensive second-line anti-TB drugs for WHO-approved TB control programmes in low- and middle-income countries. PMID- 15333268 TI - A cluster-randomised trial of screening for language disorders in toddlers. AB - OBJECTIVE: To assess the screening performance of a specific language-screening instrument at 18 and 24 months of age and to assess its effect on the early detection and prognosis of language delay. DESIGN: Child health care physicians were randomised to the intervention group, in which specific language screening was conducted twice (at age 18 months and 24 months), or to the control group (usual care). The specific screening instrument consisted of a uniform set of questions for the parents and test elements for the child, with scaled scores to assess responses. SETTING: Child health care in the Netherlands and referral of screen-positive children. SUBJECTS: 5734 children in the intervention group and 4621 in the control group. MAIN OUTCOME MEASURES: Test characteristics and disorders at 24 months, and confirmed diagnoses of a language disorder before 36 months in both groups. Gold standard based on reports of parents, specialists and expert panel. Prognosis estimated from two diagnostic language development performance scores at 36 months (in questionnaire). RESULTS: In the intervention group, 3147 of the 5734 children (55%) were screened with the specific screening instrument and 73 of the screened children (2.3%) were screen-positive. Of the screen-positive children, 41 (55%) had confirmed language delay (diagnostic assessment and/or reported treatment). The estimated sensitivity of the test ranged between 24-52% depending on the severity of language disorders. The prevalence of language disorders in three-year olds was estimated to be 2.4-5.3%. In the intervention group, 1.25-2 times more children with language delay had been diagnosed before 36 months. The assessment of language development at 36 months showed no statistically significant differences between the intervention and the control groups. CONCLUSIONS: The inclusion of a specific language screening instrument in child health centre activities resulted in the earlier detection of children with language delay. Short-term health benefits could not be demonstrated. Large-scale introduction cannot be recommended on the basis of this information alone. PMID- 15333269 TI - Modelling the impact of detecting and treating ductal carcinoma in situ in a breast screening programme. AB - OBJECTIVES: Screening has substantially increased the detection of carcinoma in situ of the breast (CIS). Opinions vary as to whether this constitutes over diagnosis or an opportunity to interrupt breast cancer's natural history. In England, incidence of invasive cancer and CIS increased in women of screening age (50-64 years), leading to a subsequent deficit in invasive incidence in women aged 65-69 years immediately beyond the invited age range. We aimed to model underlying incidence of invasive cancer and CIS expected in the absence of screening, and to quantify the likely relative contributions of their early detection to the observed reduction in invasive cancer in women of postscreening age. SETTING: UK NHS breast screening programme in England. METHODS: Poisson regression modelling was used to establish the underlying incidence of invasive and in situ cancers in the absence of screening. We then estimated age- and year specific excess detection rates attributable to screening. Applying these to population figures we estimated conservatively the relative contributions of early diagnosis of CIS and invasive cancer at 50-64 years of age to the subsequent deficit in invasive cancer in women beyond invitation age (65-69 years), for screening early in the programme and at steady state. RESULTS: Our model estimated a 1.6% annual increase in incidence, giving an estimated deficit of 4.22 invasive cancers per 10,000 women aged 65-69 years in 1996. Carcinoma in situ contributed 13-17% to the deficit, assuming a mean six year lead time and 75 100% progression to invasive cancer. At steady state, with current screening performance and with lead times of 3-4 years (invasive cancer) and 6-9 years (CIS), invasive incidence might be reduced by 5-6 cancers per 10,000 women aged 65-69 years in 2010 (15-20% of underlying incidence), CIS contributing 20-40%. DISCUSSION: The longer lead time associated with CIS attenuates the impact its early detection has on subsequent invasive incidence. At steady state screening, its detection contributes significantly to the deficit in invasive incidence. Our results suggest that, cancer for cancer, there may be just as much benefit in detecting and treating a case of CIS as there is in treating a case of invasive cancer. PMID- 15333270 TI - Number needed to screen: lives saved over 20 years of follow-up in mammographic screening. AB - OBJECTIVE: To estimate the number needed to screen with mammography to save one life, based on a stated amount of screening activity and long-term follow-up for breast cancer death. SETTING: A randomised controlled trial of mammographic screening for breast cancer, with 77,080 women invited to screening and 55,985 not invited. The invited group was offered screening for seven years. Follow-up continued for a total of just over 20 years. METHODS: Number needed to screen for seven years to save one life over 20 years was calculated by dividing the number screened (not the number invited) by the total number of lives saved. Similarly, we calculated the number of mammographic examinations required to save one life. RESULTS: We estimate that the number of women needed to screen for seven years to save one life over 20 years is 465 (95% CI 324-819). The number of mammographic examinations needed to save one life was 1499 (95% CI 1046-2642). CONCLUSIONS: The number needed to screen to save one life is smaller than has been reported in the past. Mammographic screening is effective in absolute terms as well as relative. Long-term follow-up allowed us to estimate the absolute benefit with greater accuracy. PMID- 15333271 TI - Surgical intervention in screen-detected patients versus symptomatic patients with breast cancer. AB - OBJECTIVES: The impact of population-based screening for breast cancer on the rate of breast-conserving surgery has not been established. We sought to evaluate whether surgical intervention in patients with screen-detected breast cancer differed from those with clinically detected tumours. SETTINGS: St Vincent's University Hospital and the BreastCheck Merrion Unit, part of the Irish National Breast Screening Programme, were the setting for the study. METHODS: A total of 902 patients referred for surgery to St Vincent's University Hospital over a four year period (2000-2003) were studied. Patients with breast cancers detected during the prevalent round of screening (n=325) were compared with patients presenting with symptomatic disease (n=577). The operative procedure, nature of axillary surgery and histopathological findings were recorded in each case. RESULTS: There was an increase in breast-conserving therapy in the screened population compared with symptomatic cases (68% screened versus 53% symptomatic; p<0.0001), with a corresponding reduction in axillary clearance rates (65% screened versus 81% symptomatic; p<0.0001). Nodal positivity was similar following correction for size in all tumours >1 cm, regardless of method of detection. Sentinel node biopsy was successfully undertaken in 39% of tumours <2 cm (T1 tumours) [corrected] in the screening population. CONCLUSIONS: The screened population was statistically more likely to have breast-conserving therapy than the symptomatic group. Sentinel node biopsy has evolved into an acceptable alternative to axillary clearance in T1 cancers, particularly in screen-detected cases. PMID- 15333272 TI - A population-based study of primary care predictors of non-attendance for cervical screening. AB - OBJECTIVE: To identify and estimate the population impact of primary care service delivery factors that independently predict non-attendance for cervical screening. SETTING: Screening records of all eligible women aged 30 years and over and resident in the former Manchester Health Authority area were analysed (n=72,613). METHODS: Practice-level and GP-level explanatory variables, along with area-level covariates, were obtained and merged to the study data set. Logistic regression models were used to identify factors associated with having no recorded history of attending NHS cervical screening services. A multivariate model was created to identify independent predictors of non-attendance with comprehensive adjustment for women's age, area-level socio-demographic factors, and other primary care factors. Attributable fraction estimates were used to assess the population impact of the independent predictors. RESULTS: Large practice size (>4,000 patients), single-handed practice, South Asian male GP, part-time GP employment status, older age and birthplace overseas, and area-level measures of deprivation and transience independently predicted non-attendance. Women born overseas and registered at larger practices were especially unlikely to have ever attended. The combined population attributable fraction estimate for the independent predictors reflecting primary care service delivery was almost 40%, and that for all variables in the final model was over 70%. CONCLUSIONS: Independent predictors of non-attendance reflecting general practice structure, workload and GP characteristics were identified. Although relative risks were modest, the collective population impact of these factors was considerable, which has implications for the implementation of informed targeting and the development of new screening services by Primary Care Trusts. PMID- 15333273 TI - Meta-analysis on the validity of pepsinogen test for gastric carcinoma, dysplasia or chronic atrophic gastritis screening. AB - AIM: To assess the validity of the measurement of pepsinogen I and II as a screening test for gastric cancer and pre-malignant lesions, namely low-grade dysplasia, both in the general population and in selected groups of patients. METHODS: A meta-analysis of sensitivity and specificity results from individual papers on the use of the pepsinogen test. An intrinsic cut-off effect was assumed and a random effect model was used for pooling. RESULTS: Forty-two data sets were included: 27 (64%) population-based screening studies (n=296,553) and 15 (36%) sets of selected individuals (n=4385). Homogenous sensitivity and diagnostic odds ratio (DOR) estimates were found in studies using both pepsinogen I levels and pepsinogen I/II ratio calculations. Pooled pairs of sensitivity and false positive rates (FPr) for pepsinogen I < or = 70; pepsinogen I/II ratio < or = 3, pepsinogen I < or =50; pepsinogen I/II ratio < or = 3, and pepsinogen I < or = 30; pepsinogen I/II ratio < or = 2, were sensitivity 77%/FPr 27%, sensitivity 68%/FPr 31%, and sensitivity 52%/FPr 84%, respectively. Positive predictive values (PPV) varied between 0.77% and 1.25%, and negative predictive values (NPV) varied between 99.08% and 99.90%. In selected groups, pooling was only possible when considering pepsinogen I < or = 70; pepsinogen I/II ratio < or = 3: giving sensitivity 57%, specificity 80%, PPV 15% and NPV 83%. As for the diagnosis of dysplasia, studies considering pepsinogen I <50; pepsinogen I/II ratio <3 obtained sensitivity 65% and specificity ranging from 74%-85%, both with NPV >95%. CONCLUSION: Pepsinogen test definition should include pepsinogen I/II ratio as consistency was obtained, both in population based studies and in selected groups for those studies that used pepsinogen I serum levels together with pepsinogen I/II ratio for screening for gastric cancer in high-incidence regions other than Japan. Further studies of this test in the management of high-risk patients seem to be worthwhile. PMID- 15333274 TI - Screening for physical and psychological illness in the British Armed Forces: I: The acceptability of the programme. AB - OBJECTIVES: To assess the response to a self-administered questionnaire and attendance of a medical centre for physical and psychological health screening. METHODS: 4500 men and women from the three services were randomly selected to receive either a full or abridged screening questionnaire. The full questionnaire included the General Health Questionnaire-12 (GHQ-12) and Post-traumatic Stress Disorder (PTSD) checklist, 15 symptoms, a self-assessed health status question and three questions on alcohol behaviour (WHO Audit). The abridged questionnaire included GHQ-4, a slightly shortened PTSD checklist and five symptoms, but excluded questions on alcohol behaviour. All 'screen-positive' and a random 'screen-negative' sample were invited to attend a medical centre. RESULTS: 67.1% of the servicemen completed a questionnaire; slightly but significantly more the abridged than the full questionnaire (4.9%, 95% confidence interval 2.3-7.4%). Of those receiving a full or abridged questionnaire, 32% and 22.5% respectively were 'screen-positives', most of the difference (7.5%) attributable to alcohol behaviour. Less than 30% of the servicemen invited to attend a medical centre accepted the invitation, even fewer during the preparation for deployment to Iraq. Those who fulfilled the criteria for PTSD, alcohol behaviour or multi criteria 'screen-positive' were more reluctant than controls to attend. CONCLUSIONS: Screening for psychological illness has little support among servicemen, perhaps because they may not wish to share concerns with a military doctor. Avoidance behaviour among those with a psychological condition may also selectively reduce willingness to attend a medical centre. Screening during pre deployment periods has even less support than at other times. PMID- 15333275 TI - Screening for physical and psychological illness in the British Armed Forces: II: Barriers to screening--learning from the opinions of Service personnel. AB - OBJECTIVE: To identify any potential barriers to the effectiveness of a military health screening programme based on the beliefs of British Service personnel. METHODS: As part of a pilot evaluation of the suitability of a new health screening questionnaire for the British Armed Forces, 73 men and women from the three Services, of various ranks and age, underwent a semi-structured interview after completing a screening questionnaire. Participants were asked about the veracity of their answers and their views regarding a screening questionnaire. Afterwards questionnaires were sent to 4496 randomly selected personnel from the three Services, which validated the main emerging themes. A constant comparative method of analysis was used to identify and categorise all ideas presented. RESULTS: The main barriers to health screening were lack of trust, perceived low quality of healthcare, and perceived lack of concern within the institution about work environments and home life. The central issue was 'confidence' in military health care provision. Screening was considered worthwhile, but many confided that they would not honestly answer some items in the questionnaire. Lack of trust in medical confidentiality, stigmatisation and fears that the process would jeopardise career prospects were stressed. Many Service personnel admitted to seeking medical help outside the Armed Forces. CONCLUSIONS: Concerns raised by Service personnel may endanger the value of a screening programme and the provision of health services. Greater emphasis needs to be placed upon gaining the confidence of those targeted for health screening. PMID- 15333276 TI - Screening for physical and psychological illness in the British Armed Forces: III: The value of a questionnaire to assist a Medical Officer to decide who needs help. AB - OBJECTIVES: To estimate the positive and negative predictive values (PPVT and NPVT), sensitivity and specificity of a full and abridged screening questionnaire of physical and psychological health, using primary care doctors' (medical officers [MOs]) assessments as to whether the servicemen needed medical help as a gold standard. METHODS: From a tri-service random sample of those who completed a questionnaire, all 'screen-positive' and an equal random sample of 'screen negative' were selected to attend their medical centre. MOs were aware that the screening was aimed at detecting psychological illness, but were blind as to the 'screen-positivity' of any serviceman. The MO completed a questionnaire that asked whether the patient needed medical help and whether s/he was previously aware of this need. RESULTS: 314 subjects were available for analysis. The PPVT was 47% (95% confidence interval [CI] 36-59%) for the full questionnaire and 48% (95% CI 36-60%) for the abridged questionnaire. Of those 'screen-positive' subjects whom the MO rated as needing help, one third had problems already known to the MO, regardless of the length of the questionnaire. The sensitivity and specificity of the full and abridged questionnaires were 43% and 74%, and 36% and 83% respectively. The PPVT did not vary greatly between health dimensions nor did selection of servicemen with very high scores. CONCLUSIONS: The use of MOs as a gold standard is important because of their central role in initiating the management of any condition uncovered by a screening programme. Using MOs as a gold standard, the validity of the screening questionnaires for physical and psychological health in the military was mediocre. PMID- 15333279 TI - Report from the CDC. Use of preventive health services by Hispanic/Latino women in two urban communities: Atlanta, Georgia and Miami, Florida, 2000 and 2001. AB - PURPOSE: During the 1990s, a 58% increase in the Hispanic/Latino population, fueled by the century's largest immigration wave and the highest fertility of any group, resulted in Hispanics becoming the largest U. S. minority group. To assess use of preventive services by Hispanics in Atlanta, Georgia, the largest Hispanic new destination, and Miami, Florida, the largest established Hispanic community in the Southeast, survey data were analyzed. METHODS: Miami-Ft. Lauderdale and Atlanta metropolitan area data from the 2000 National Health Interview Survey (NHIS) and from anonymous surveys conducted at health festivals in Miami and Atlanta in 2001 were analyzed. RESULTS: Female non-Hispanic white and black NHIS respondents were more likely than Hispanic counterparts to report annual household income >$20,000 (77.3%, 70.8% vs. 67.7%), usual source of healthcare (61.5%, 56.4% vs. 50.2%), or ever having had Pap screening (88.8%, 86.7% vs. 80.7%) or oral contraceptive use (55.7%, 59.7% vs. 33.7%). Miami-Ft. Lauderdale Hispanics were less likely than Atlanta respondents to be monolingual Spanish speakers, to lack usual source of healthcare, or to have less than 12 years of education. Of 295 female health festival respondents, the 160 Miami participants were more likely than Atlanta participants to have health insurance, monthly income >$1000, and prior Pap screening (p < 0.01) but less likely to have used contraception (p = 0.07). Most Hispanics felt they had inadequate healthcare; 15.0% reported being denied healthcare because of inability to pay. CONCLUSIONS: Low income, uninsured status, and language barriers were associated with lower use of preventive services among Hispanics in these Southeastern communities, particularly Atlanta, a new destination. PMID- 15333280 TI - Toward optimal health: the experts discuss eating disorders. PMID- 15333281 TI - Bulimia: medical complications. AB - Bulimia nervosa is a common eating disorder that predominantly affects young women. There are three main models of purging in bulimia. Resulting medical complications are related to the particular mode and frequency of purging. Commonly, there are oral and gastrointestinal complications along with serious electrolyte and endocrine complications. The majority of the medical complications of bulimia nervosa are treatable if diagnosed in a timely fashion. Some of these patients require inpatient hospitalization, and others can be managed along a continuum of outpatient care. The American Psychiatric Association has comprehensive treatment guidelines for the management of bulimia. Primary care physicians and gynecologists need to be familiar with this disorder and its medical implications. PMID- 15333282 TI - Predictors of condom-related attitudes among at-risk women. AB - BACKGROUND: Negative attitudes toward using male condoms tend to be associated with higher rates of sexual risk. Little has been written about the factors that influence women's attitudes toward condom use, and this has implications for HIV intervention efforts. METHODS: Two hundred fifty adult women considered to be at risk based on demographic and family characteristics and residing in the Atlanta, Georgia, metropolitan area were interviewed between August 1997 and August 2000. Street outreach efforts were used to identify potential study participants, with further expansion of the sample done via targeted sampling and ethnographic mapping procedures. RESULTS: Women held ambivalent to weakly positive attitudes toward condoms. Multivariate analysis revealed that five factors were associated with a greater frequency of engaging in seven specific types of sexual activities known to be associated with HIV transmission: condom-related attitudes (negative condom attitudes), marital status (being married vs. other marital status classification), religiosity (lesser), childhood physical abuse (greater), and the amount of illegal drug use (greater). Four significant predictors of condom attitudes were also identified: age (better condom attitudes among younger women), childhood neglect (more conductive condom attitudes among nonneglected women), self-esteem (more self-esteem = more favorable condom attitudes), and the number of drug problems experienced (more drug problems = more negative condom attitudes). CONCLUSIONS: The more negative at-risk women's attitudes were regarding condom use, the more often they tended to engage in risky sex. Specific backgrounds and characteristics of the women were associated with greater/lesser condom use. To increase condom use, programs should consider targeting specific types of women as well as their specific attitudes toward condom use. PMID- 15333283 TI - Women, the Internet, and sexually transmitted infections. AB - OBJECTIVE: To assess the relationship between Internet use and sexual risk behavior among women. METHODS: A cross-sectional survey, with 64 items, seeking sex partners online was posted on the Internet. Respondents were targeted through e-mail list serves and bulletin board postings. However, the majority of responses were gathered after national media coverage of the survey. RESULTS: Of the 4444 eligible participants, age >18, from North America who completed the online survey, 29% (n = 1276) were women. Of the 1276 women, 544 (43%) reported having sex with a person they first met on the Internet. These women were more frequently white, older, and more experienced with testing for sexually transmitted infections (STI) than women who never had a sex partner they first met online. They also had a higher number of total lifetime partners and more frequently used condoms. However, these women also reported high rates of STI, are not regularly using condoms, and are engaging in anal, oral, and vaginal sex with Internet partners. Many of the women say they would appreciate receiving STI prevention information via e-mail, and those with Internet partners more frequently stated they would participate in chat room discussions about prevention (p < 0.02). CONCLUSIONS: The Internet is a flourishing sex venue, and women too are using this new vehicle to seek out sex partners. The wide pool of potential sex partners found online coupled with ease of travel and an increase in partners has the potential to spread an STI or HIV with greater efficiency than ever before imagined. Although women with Internet partners may engage in protective behavior more frequently than women with no Internet partners, they also engage in higher risk behaviors. This population, therefore, needs to be recognized and targeted with STI and HIV education and prevention efforts. PMID- 15333284 TI - Does hormonal status influence the clinical presentation of acute coronary syndromes in women? AB - BACKGROUND: Previous studies suggest that the clinical presentation of acute coronary syndromes (ACS) may differ between women and men. It is not known if different clinical presentations may be explained by hormonal status in women with ACS. Our objective was to compare the clinical presentation of ACS between premenopausal (PRE) women and post-menopausal women with hormone replacement therapy (HRT) and without (POST). METHODS: This was a prospective study of consecutive women admitted with a diagnosis of ACS (myocardial infarction [MI] or unstable angina). All women answered a detailed questionnaire that included a list of 27 clinical symptoms. Symptom results were adjusted for age and current coronary event diagnosis. RESULTS: Seventy-three Caucasian women were studied. No differences were found in terms of the frequency of reported typical symptoms of ACS between PRE (n = 23), HRT (n = 32), and POST (n = 18). However, PRE more often reported atypical chest symptoms than HRT and POST women (57% vs. 31% vs. 22%, PRE vs. HRT vs. POST, respectively, p = 0.05). HRT and POST women experienced substernal chest pain more frequently than PRE (44% vs. 78% vs. 83%, p = 0.03). In contrast, PRE more frequently tended to experience chest pressure (57% vs. 31% vs. 39%, p = 0.2) or chest pain in other locations (22% vs. 3% vs. 6%, p = 0.06). HRT and POST groups reported more frequent indigestion-like pain/discomfort (22% vs. 50% vs. 56%, p = 0.04) and midback pain (35% vs. 63% vs. 72%, p = 0.04) during ACS compared with PRE women. POST experienced sudden fatigue more frequently than PRE and HRT (61% vs. 53% vs. 89%, p = 0.03). CONCLUSIONS: Our results suggest that almost all women admitted with ACS experienced typical chest symptoms but frequently reported both typical and atypical symptoms. Independently of age, atypical chest symptoms occurred more frequently in premenopausal women than in postmenopausal women with or without HRT. PMID- 15333285 TI - Gender differences in the practice of adult primary care physicians. AB - OBJECTIVE: This study investigates how physician gender affects reactions to six model patients. METHODS: Telephone interviews with 3205 internists and family or general physicians were completed between 1996 and 1997 for the Community Tracking Study. Physicians responded to six vignettes describing model patients with presentations designed to have multiple appropriate treatment plans: a 50 year-old man with a 1-month history of exertional chest pain who may need a referral to a cardiologist, a 60-year-old man with a normal digital rectal examination (DRE) who may benefit from a prostate-specific antigen (PSA) test, a 40-year-old married woman with vaginal itching and discharge who may benefit from an office visit, a 60-year-old man with symptoms of benign prostatic hypertrophy (BPH) who may benefit from a urological consultation, a 35-year-old man with back pain and a new left footdrop who may benefit from an MRI, and a 50-year-old man with elevated cholesterol and no other cardiac risk factors who may benefit from cholesterol-lowering agents. RESULTS: Female physicians are significantly more likely than males to refer a patient with BPH to a urologist (37.5% vs. 24.9%, p < 0.001). Male physicians are significantly more likely to recommend that a woman with vaginal itching and discharge have an office visit (52.7% vs. 40.6%, p < 0.001). Male physicians recommend cholesterol-lowering agents slightly more often than women physicians (39.4% vs. 36.4%, p < 0.03) and recommend a PSA test more often than female physicians (73.1% vs. 64.4%, p < 0.001). Neither physician attributes, practice characteristics, referral patterns, nor geographical traits account for the disparity between male and female physicians in the treatment of BPH. Approximately 40% of the gender gap in treatment of vaginal itching and discharge can be attributed to physician attributes. The association between provider gender and the decision to prescribe cholesterol-lowering agents and the association between provider gender and recommending a PSA test are explained by physician attributes. CONCLUSIONS: Gender differences in treatment seem to appear most strongly for genital-specific conditions. The results may suggest that physicians use fewer resources to treat the genital-specific conditions of patients who share their sex. PMID- 15333286 TI - Prevalence of physician-diagnosed systemic lupus erythematosus in the United States: results from the third national health and nutrition examination survey. AB - OBJECTIVE: To determine the prevalence of physician-diagnosed systemic lupus erythematosus (SLE) in a national population-based sample in the United States. METHODS: Data from the Third National Health and Nutrition Examination Survey (NHANES III) were used to estimate the prevalence of self-reported physician diagnosed SLE. Adult participants (age > or = 17; sample n = 20,050) were asked if they had been diagnosed with SLE by a physician. All medications currently being taken by survey participants were recorded. Two definitions were used to classify participants with SLE: self-reported physician diagnosis and self reported physician diagnosis and a current prescription for antimalarials, corticosteroids, or other immunosuppressive medications. RESULTS: The prevalence of SLE in adults age > or = 17 based on self-reported physician diagnosis was 241 per 100,000 (95% confidence interval [CI] 130-352). The prevalence of SLE in adults age > or = 17 based on self-reported physician diagnosis and current prescription for antimalarials, corticosteroids, or immunosuppressive medications was 53.6 per 100,000 (95% CI 12.2-95.0). Among adult women, the prevalence of treated SLE was 100 per 100,000 (95% CI 19.8-179.3). CONCLUSIONS: Projecting a prevalence of 100 per 100,000 to the population of the United States, approximately 108,300 adult women had a self-reported physician diagnosis of SLE and were receiving specific treatment in 2000. This estimate is a reasonable lower boundary, as it does not include undiagnosed persons or those not being treated with antimalarials, corticosteroids, or immunosuppressive medications. PMID- 15333287 TI - Changing trends in clinical AIDS presentations and survival among HIV-1-infected women. AB - OBJECTIVES: To profile trends of clinical AIDS-defining illness (ADI) among a cohort of human immunodeficiency virus (HIV)-infected women over a 12-year period. METHODS: In a prospective evaluation of AIDS clinical presentation in the Women and Infants Transmission Study (WITS), 2255 subjects were enrolled and followed between December 1989 and June 2002 (total, 4993 person-years). Data on clinical AIDS presentation of 140 (6.2%) HIV-seropositive subjects were evaluated across three calendar periods corresponding to the use of different therapy regimens. Incidence rates (per 1000 woman-years) for AIDS and specific ADIs were compared between periods using Poisson regression methods. RESULTS: Incidence rates of AIDS, Mycobacterium tuberculosis, recurrent bacterial pneumonia, herpes simplex disease, esophageal/bronchial candidiasis, wasting syndrome, and neurological diseases have showed significant downward trends. Among women with ADI, the frequency of either esophageal or bronchial candidiasis as initial ADI showed an increasing trend (p(trend) = 0.03), whereas a decrease in proportion of cases with nontuberculosis mycobacterial infection (P(trend) = 0.05) was observed over the same periods. In the multivariate analysis, both the CD4+ lymphocyte count and HIV-1 RNA at the time of diagnosis were independently associated with survival after AIDS. Highly active antiretroviral therapy (HAART) was associated with a 70% reduction in progression to death following AIDS. CONCLUSIONS: Temporal changes in the incidence and clinical presentations in HIV-positive women in our cohort reflect an increased use of HAART that may have a differential effect on reduction in the risk of ADIs. These illnesses, although considerably less frequent in recent years, are still important contributors to morbidity in HIV-positive women. PMID- 15333288 TI - Depressive symptom burden as a barrier to screening for breast and cervical cancers. AB - BACKGROUND: Depression has been associated with lower cancer screening rates in some studies. We examined whether a higher depressive symptom burden presented a barrier to subsequent mammography and Papanicolaou (Pap) smear testing. METHODS: Study of Women's Health Across the Nation (SWAN) is a cohort study of 3302 community-dwelling women. At baseline, participants were 42-52 years old, had no surgical removal of the uterus or both ovaries, no current use of hormones that affect the ovaries, and at least one menses in the previous 3 months. SWAN data spanned 4 years. Repeated measures logistic models determined odds of mammography and of Pap screening in the year following depressive symptom burden, as determined by Center for Epidemiological Studies Depression score (CES-D). The models controlled for age, race/ethnicity, health insurance, medical history and use, smoking, obesity, and socioeconomic status. RESULTS: At baseline, 75.6% (2493 of 3297) had a low depressive symptom burden (CES-D score < 16, referent), 9.5% (312 of 3297) had a moderate burden (CES-D 16-20), and 14.9% (492/3297) had a high burden (CES-D > or = 21). Women with a high depressive symptom burden had, in the subsequent year, significantly lower odds of mammography (OR 0.84, 95% CI 0.73-0.97) but not Pap smear (OR 0.88, 95% CI 0.76-1.03). There was not a significant dose-response relationship between depressive symptom burden and screening. CONCLUSIONS: The presence of a high depressive symptom burden is a modest independent risk factor for lack of subsequent mammography. Ensuring that depressed patients receive regular cancer screening services is important. PMID- 15333289 TI - Depression as a risk factor for underuse of mammography. PMID- 15333290 TI - Complex clinical, legal, and ethical issues of pregnant and postpartum women as subjects in clinical trials. PMID- 15333293 TI - Changes in Smad expression and subcellular localization in bleomycin-induced pulmonary fibrosis. AB - Administration of bleomycin (BM) produces inflammation and fibrosis of the lung in humans and experimental animals. The molecular defects by which BM induces these pathological effects have not been studied in detail. We studied the expression of Smad family proteins, key molecules involved in mediating transforming growth factor (TGF)-beta signaling from the cell membrane to the nucleus, during the early and late phases of BM-induced fibrogenesis. Pulmonary fibrosis was induced in male Sprague-Dawley rats by a single intratracheal injection (1.5 units) of BM. Control rats received saline. Rats were killed at 3, 5, 7, 14, and 28 days after BM, cytosolic and nuclear proteins were extracted and isolated from lung tissues, and Smad proteins were probed with specific antibodies. In BM-exposed lung tissue, compared with control, Smad3 decreased persistently in the cytosol and increased transiently in the nucleus. There was a persistent increase in phosphorylation and nuclear accumulation of Smad2/3. Smad4 was increased transiently in both the cytosol and nucleus. A significant and progressive decrease in the expression of Smad7, the endogenous inhibitor of TGF beta/Smad signaling, was observed after BM instillation. Collectively, our results indicate that an imbalance between agonistic Smads2-4 and antagonistic Smad7 may result in the unchecked activation of an autocrine TGF-beta loop, which contributes to the pathogenesis of BM-induced pulmonary fibrosis. PMID- 15333294 TI - TREND: an important step, but not enough. PMID- 15333295 TI - Tobacco control? PMID- 15333296 TI - Moving beyond randomized controlled trials. PMID- 15333297 TI - Making America fit and trim--steps big and small. PMID- 15333298 TI - Walking the talk: Fit WIC wellness programs improve self-efficacy in pediatric obesity prevention counseling. AB - Six sites of the California Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) participated in a staff wellness pilot intervention designed to improve staff self-efficacy in counseling WIC clients about childhood overweight. A pre-post test design with intervention and control groups was used; outcome measures included staff perceptions of the intervention's effects on the workplace environment, their personal habits and health beliefs, and their counseling self-efficacy. Intervention site staff were more likely to report that the workplace environment supported their efforts to make healthy food choices (P <.001), be physically active (P <.01), make positive changes in counseling parents about their children's weight (P <.01), and feel more comfortable in encouraging WIC clients to do physical activities with their children (P <.05). PMID- 15333299 TI - Estimating deaths attributable to obesity in the United States. AB - Estimates of deaths attributable to obesity in the United States rely on estimates from epidemiological cohorts of the relative risk of mortality associated with obesity. However, these relative risk estimates are not necessarily appropriate for the total US population, in part because of exclusions to control for baseline health status and exclusion or underrepresentation of older adults. Most deaths occur among older adults; estimates of deaths attributable to obesity can vary widely depending on the assumptions about the relative risks of mortality associated with obesity among the elderly. Thus, it may be difficult to estimate deaths attributable to obesity with adequate accuracy and precision. We urge efforts to improve the data and methods for estimating this statistic. PMID- 15333300 TI - Feasibility and benefits of a parent-focused preschool child obesity intervention. AB - OBJECTIVES: This field study tested the feasibility and benefits of a program to promote 6 targeted parental behaviors to prevent obesity in children served by the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). METHODS: Two WIC sites participated in a nonrandomized, controlled 1-year prospective study to assess parents' self-reported behavior changes. RESULTS: Statistical analyses of preintervention and postintervention assessments of parental behavior demonstrated significant changes in 2 behaviors: frequency of offering the child water and frequency of engaging in active play with the child. In both cases, the intervention proved effective in increasing the desired behavior. CONCLUSIONS: The findings demonstrate the feasibility of changing parental behaviors through multidimensional education in a WIC clinic setting. PMID- 15333301 TI - Childhood obesity in New York City elementary school students. AB - OBJECTIVES: We estimated overweight and obesity in New York City elementary school children. METHODS: A multistage cluster sample of New York City public elementary school children was selected. Nurses measured children's height and weight and used a standard protocol to determine body mass index (BMI). Demographic information was obtained from official school rosters. Overweight and obese were defined as BMI-for-age at or above the 85th and 95th percentiles, respectively. RESULTS: Of 3069 sampled students, 2681 (87%) were measured. The prevalence of overweight was 43% (95% confidence interval [CI] = 39%, 47%), more than half of whom were obese. Overall prevalence of obesity was 24% (95% CI = 21%, 27%), with at least 20% obesity in each grade, including kindergarten. Hispanic children had significantly higher levels (31%; 95% CI = 29%, 34%) than Black (23%; 95% CI = 18%, 28%) or White children (16%; 95% CI = 12%, 20%). Asian children had the lowest level of obesity among all racial/ethnic groups (14.4%, 95% CI = 10.9, 18.7). CONCLUSIONS: Obesity among public elementary school children in New York City is an important public health issue. Particularly high levels among Hispanic and Black children mirror national trends and are insufficiently understood. PMID- 15333302 TI - Physical education in elementary school and body mass index: evidence from the early childhood longitudinal study. AB - OBJECTIVES: We examined the effect of physical education instruction time on body mass index (BMI) change in elementary school. METHODS: We examined data from a national sample of 9751 kindergartners in the United States who were reported on for 2 years. We used a difference-in-differences approach to examine the effect of an increase in physical education instruction time between kindergarten and first grade on the difference in BMI change in the 2 grades, using the same child as the control. RESULTS: One additional hour of physical education in first grade compared with the time allowed for physical education in kindergarten reduces BMI among girls who were overweight or at risk for overweight in kindergarten (coefficient = -0.31, P <.001) but has no significant effect among overweight or at-risk-for-overweight boys (coefficient = -0.07, P =.25) or among boys (coefficient = 0.04, P =.31) or girls (coefficient = 0.01, P =.80) with a normal BMI. CONCLUSIONS: Expanding physical education programs in schools, in the form in which they currently exist, may be an effective intervention for combating obesity in the early years, especially among girls. PMID- 15333303 TI - An environmental intervention to promote lower-fat food choices in secondary schools: outcomes of the TACOS Study. AB - OBJECTIVES: We evaluated an environmental intervention intended to increase sales of lower-fat foods in secondary school cafeterias. METHODS: Twenty secondary schools were randomly assigned to either an environmental intervention or a control group for a 2-year period. The intervention increased the availability of lower-fat foods and implemented student-based promotions. RESULTS: A steeper rate of increase in sales of lower-fat foods in year 1 (10% intervention vs -2.8% control, P =.002) and a higher percentage of sales of lower-fat foods in year 2 (33.6% intervention vs 22.1% control, P =.04) were observed. There were no significant changes in student self-reported food choices. CONCLUSIONS: School based environmental interventions to increase availability and promotion of lower fat foods can increase purchase of these foods among adolescents. PMID- 15333304 TI - Trends in blood lead levels among children enrolled in the Special Supplemental Nutrition Program for Women, Infants, and Children from 1996 to 2000. AB - We analyzed data from the Wisconsin Childhood Lead Poisoning Prevention Program to examine the distribution of and trends in elevated blood lead levels among WIC enrolled children from 1996 until 2000. Higher blood lead levels were seen among WIC-enrolled children, and although not statistically significant, the rate of blood lead level decline among WIC-enrolled children was greater than among non WIC-enrolled children. PMID- 15333305 TI - Perceived need for workplace accommodation and labor-force participation in Canadian adults with activity limitations. AB - We examined how perceived need for workplace accommodation affects labor-force participation in people with disabilities. We analyzed a Canadian survey with structural equation modeling to test a model incorporating activity limitations and perceived need for workplace accommodations. The results suggested that the effect of upper- and lower-body activity limitation on labor-force participation was mediated by perceived need for workplace accommodations. Thus, the provision of adequate workplace accommodations could enhance labor-force participation in people with disabilities. PMID- 15333306 TI - Mortality in 13 French cities during the August 2003 heat wave. AB - We observed the daily trend in mortality rates during the 2003 heat wave in 13 of France's largest cities. Mortality data were collected from July 25 to September 15 each year from 1999 through 2003. The conjunction of a maximum temperature of 35 degrees C and a minimum temperature of 20 degrees C was exceptional in 7 cities. An excess mortality rate was observed in the 13 towns, with disparities from +4% (Lille) to +142% (Paris). PMID- 15333307 TI - Municipal heat wave response plans. AB - Approximately 400 people die from extreme heat each year in the United States, and the risk of heat waves may increase as a result of global climate change. Despite the risk of heat-related morbidity and mortality, many cities lack written heat response plans. In a review of plans from 18 cities at risk for heat related mortality, we found that many cities had inadequate or no heat response plans. This is an important area for further investigation and government attention. PMID- 15333308 TI - Alcohol and opiate withdrawal in US jails. AB - We sought to estimate the number of arrestees at risk for inadequately treated drug and alcohol withdrawal in US jails. We used Arrestee Drug Abuse Monitoring Program data to estimate prevalence rates of alcohol and opiate dependence. Our results revealed rates of alcohol and opiate dependency among arrestees of approximately 12% and 4%, respectively; only 28% of jail administrators reported that their institutions had ever detoxified arrestees. Inadequately treated drug and alcohol withdrawal in US jails appears widespread. Our data raise important ethical and constitutional questions. PMID- 15333309 TI - Changes in diet quality of American preschoolers between 1977 and 1998. AB - OBJECTIVES: We determined diet quality trends among nationally representative samples of preschoolers between 1977 and 1998. METHODS: Adjusted diet quality index scores, overall intake, and tertiles of total score were compared for combined samples and 2 age groups using t tests with Bonferroni correction; surveys used were the US Department of Agriculture's National Food Consumption Survey 1977-1979 (n = 2342), Continuing Survey of Food Intake by Individuals (CSFII) 1989-1991 (n = 858), and CSFII 1994-1996 and 1998 (n = 5355). RESULTS: Total scores increased slightly. Consumption of grains, fruits, and vegetables improved while added sugar and juice intake worsened. CONCLUSIONS: Diet quality improved marginally since 1977. Consumption of fruits and vegetables needs to be increased and that of total and saturated fat, juice, and added sugar decreased. PMID- 15333310 TI - The relation of breastfeeding and body mass index to asthma and atopy in children: a prospective cohort study to age 6 years. AB - OBJECTIVES: We investigated the relationship between breastfeeding, asthma and atopy, and child body mass index (BMI). METHODS: From a prospective birth cohort (n = 2860) in Perth, Western Australia, 2195 children were followed up to age 6 years. Asthma was defined as doctor-diagnosed asthma and wheeze in the last year, and atopy was determined by skin prick test of 1596 children. Breastfeeding, BMI, asthma, and atopy were regressed allowing for confounders and the propensity score for overweight. RESULTS: Using fractional polynomials, we found no association between breastfeeding and overweight. Less exclusive breastfeeding was associated with increased asthma and atopy, and BMI increased with asthma. CONCLUSIONS: Less exclusive breastfeeding leads to increases in child asthma and atopy and a higher BMI is a risk factor for asthma. PMID- 15333311 TI - Improvements in heart health behaviors and reduction in coronary artery disease risk factors in urban teenaged girls through a school-based intervention: the PATH program. AB - OBJECTIVES: We sought to assess the effects of a school-based intervention program on cardiovascular disease risk factors in urban girls. METHODS: We compared heart health knowledge, health behaviors, cardiovascular risk factors, and physical fitness among a group of 442 multiethnic teenaged girls (310 experimental participants vs 132 control participants). Testing was conducted before and after a 12-week program of vigorous exercises integrated with lectures and discussions on diet, exercise, stress, and smoking. RESULTS: Significant differences in body fat, systolic and diastolic blood pressure, heart health knowledge, and whether breakfast was eaten were observed between experimental participants and control participants. CONCLUSIONS: An integrated program of exercise and heart health-related lectures and discussions had a beneficial effect on health knowledge, health behaviors, and onset of risk factors for coronary artery disease among urban girls. PMID- 15333312 TI - Trends in overweight among adolescents living in the poorest and richest regions of Brazil. AB - OBJECTIVES: We assessed changes in body mass index (BMI) among Brazilian adolescents. METHODS: In 1975, 1989, and 1997, we conducted household surveys of the weights and statures of a probabilistic sample of about 50,000 Brazilian adolescents aged 10 to 19 years. Weighted prevalences were calculated and an analysis was performed with the sample design taken into account. RESULTS: Adolescents of rich (southeast) and poor (northeast) regions showed a substantial increase in BMI. In the southeast, the prevalence of overweight, defined by international age- and gender-specific BMI cutoffs, for both genders reached 17% in 1997, whereas in the northeast, the prevalence tripled, reaching 5% among boys and 12% among girls. Older girls living in urban areas in the southeast showed a decrease in prevalence from 16% to 13% in the latter 2 surveys. For all boys and for young girls, the BMI values for the 85th percentile in 1997 were much higher than the 95th percentile values in 1975. CONCLUSIONS: BMI increased dramatically in Brazilian adolescents, mainly among boys; among older girls from the richest region, the prevalence of overweight is decreasing. PMID- 15333313 TI - Barriers to buying healthy foods for people with diabetes: evidence of environmental disparities. AB - OBJECTIVES: A community coalition compared the availability and cost of diabetes healthy foods in a racial/ethnic minority neighborhood in East Harlem, with those in the adjacent, largely White and affluent Upper East Side in New York City. METHODS: We documented which of 173 East Harlem and 152 Upper East Side grocery stores stocked 5 recommended foods. RESULTS: Overall, 18% of East Harlem stores stocked recommended foods, compared with 58% of stores in the Upper East Side (P <.0001). Only 9% of East Harlem bodegas (neighborhood stores) carried all items (vs 48% of Upper East Side bodegas), though East Harlem had more bodegas. East Harlem residents were more likely than Upper East Side residents (50% vs 24%) to have stores on their block that did not stock recommended foods and less likely (26% vs 30%) to have stores on their block that stocked recommended foods. CONCLUSIONS: A greater effort needs to be made to make available stores that carry diabetes-healthy foods. PMID- 15333314 TI - Replacing fats and sweets with vegetables and fruits--a question of cost. AB - OBJECTIVES: We examined the association between diet quality and estimated diet costs. METHODS: Freely chosen diets of 837 French adults were assessed by a dietary history method. Mean national food prices for 57 foods were used to estimate diet costs. RESULTS: Diets high in fat, sugar, and grains were associated with lower diet costs after adjustment for energy intakes, gender, and age. For most levels of energy intake, each additional 100 g of fats and sweets was associated with a 0.05-0.40 per day reduction in diet costs. In contrast, each additional 100 g of fruit and vegetables was associated with a 0.18-0.29 per day increase in diet costs. CONCLUSIONS: Diets high in fats and sweets represent a low-cost option to the consumer, whereas the recommended "prudent" diets cost more. PMID- 15333315 TI - Sociodemographic and geographic correlates of meeting current recommendations for physical activity in middle-aged French adults: the Supplementation en Vitamines et Mineraux Antioxydants (SUVIMAX) Study. AB - OBJECTIVE: We evaluated the characteristics of French subjects meeting current public health recommendations for physical activity. METHODS: We assessed leisure time physical activity cross-sectionally in 7404 adults aged 45 to 68 years with applied logistic regression models. RESULTS: Meeting the recommended physical activity levels was more likely in subjects aged 60 years and older and in women with higher education levels or living in rural areas and was less likely in smokers. No association was found with time spent watching television. The contribution of vigorous activity to total time spent being active was approximately 2 times higher in subjects meeting recommendations. CONCLUSIONS: Participation in some vigorous activity may be viewed as a "facilitator" to attain physical activity recommendations. Relationships with physical environment variables in Europe need further investigation. PMID- 15333316 TI - Body mass index, physical activity, and the risk of decline in overall health and physical functioning in late middle age. AB - OBJECTIVES: We examined the relation between body mass index, exercise, overall health, and physical functioning. METHODS: We studied 7867 adults aged 51 to 61 years in 1992 to 1996. Adjusted relative risks for health decline and new physical difficulties were determined with logistic regression. RESULTS: Overweight and obesity were independently associated with health decline (adjusted relative risk [ARR] = 1.29 and 1.36) and development of a new physical difficulty (ARR = 1.27 and 1.45). Regular exercise significantly reduced the risk of health decline and development of a new physical difficulty, even among obese individuals. CONCLUSIONS: Maintaining ideal body weight is important in preventing decline in overall health and physical functioning. However, regular exercise can reduce the risk of health decline even among individuals who cannot achieve ideal weight. PMID- 15333317 TI - Urban sprawl and risk for being overweight or obese. AB - OBJECTIVES: I examined the association between urban sprawl and the risk for being overweight or obese among US adults. METHODS: A measure of urban sprawl in metropolitan areas was derived from the 2000 US Census; individual-level data were obtained from the Behavioral Risk Factor Surveillance System. I used multilevel analysis to assess the association between urban sprawl and obesity. RESULTS: After I controlled for gender, age, race/ethnicity, income, and education, for each 1-point rise in the urban sprawl index (0-100 scale), the risk for being overweight increased by 0.2% and the risk for being obese increased by 0.5%. CONCLUSIONS: The current obesity epidemic has many causes, but there is an association between urban sprawl and obesity. PMID- 15333318 TI - A potential natural treatment for attention-deficit/hyperactivity disorder: evidence from a national study. AB - OBJECTIVES: We examined the impact of relatively "green" or natural settings on attention-deficit/hyperactivity disorder (ADHD) symptoms across diverse subpopulations of children. METHODS: Parents nationwide rated the aftereffects of 49 common after-school and weekend activities on children's symptoms. Aftereffects were compared for activities conducted in green outdoor settings versus those conducted in both built outdoor and indoor settings. RESULTS: In this national, nonprobability sample, green outdoor activities reduced symptoms significantly more than did activities conducted in other settings, even when activities were matched across settings. Findings were consistent across age, gender, and income groups; community types; geographic regions; and diagnoses. CONCLUSIONS: Green outdoor settings appear to reduce ADHD symptoms in children across a wide range of individual, residential, and case characteristics. PMID- 15333319 TI - Length of hospital stays among obese individuals. AB - OBJECTIVES: We examined lengths of hospital stay among individuals categorized according to weight status. METHODS: We used data from the First National Health and Nutrition Examination Survey Epidemiologic Followup Survey to estimate length of-stay differences. RESULTS: Individuals with body mass indexes (BMIs) of 35 kg/m2 or above, those with BMIs of 30 to 34 kg/m2, and those with BMIs of 25 to 29 kg/m2 had crude length-of-stay rates greater than those of normal-weight individuals. Association between BMI and length of stay varied over time. CONCLUSIONS: Obese individuals experience longer hospital stays than normal weight individuals. PMID- 15333320 TI - Thinness among young Japanese women. AB - OBJECTIVES: We described changes in body mass index (BMI) and the prevalence of thinness among young Japanese women (aged 15-29 years) from 1976 to 2000 by reanalyzing the nationwide data in the National Nutrition Survey, Japan. METHODS: We used height and weight data sets for 30 903 nonpregnant, nonlactating women during the 25-year period. We calculated the mean values of BMI and the prevalence of thinness for 3 age groups (15-19, 20-24, and 25-29 years of age). RESULTS: Changes in BMI per 10 years were -0.17 kg/m2, -0.22 kg/m2, and -0.34 kg/m2 for each age group, respectively. Extreme thinness (BMI < 17 kg/m2) increased from 2.4% in 1976-1980 to 4.2% in 1996-2000. CONCLUSIONS: Further studies regarding topics such as increased smoking prevalence are needed to identify the underlying causes of increasing thinness. PMID- 15333321 TI - Predictors of help seeking among Connecticut adults after September 11, 2001. AB - OBJECTIVES: We conducted a population-based telephone survey in an attempt to determine correlates of formal and informal help seeking after September 11, 2001. METHODS: Between October 15 and December 31, 2001, 1774 Connecticut Behavioral Risk Factor Surveillance System respondents were asked questions directly related to their experiences of September 11. RESULTS: Multivariate logistic regression analyses showed that receipt of formal help was predicted by sleep problems, close association with a victim, reports of increased smoking or drinking, and receipt of informal help. Age, gender, reports of 1 or more problems, and formal help seeking predicted receipt of informal help. CONCLUSIONS: Public health planning and bioterrorism preparedness should include programs addressing increased smoking and drinking, sleep problems, and bereavement in the wake of disasters. PMID- 15333322 TI - Associations between early-adolescent substance use and subsequent young-adult substance use disorders and psychiatric disorders among a multiethnic male sample in South Florida. AB - OBJECTIVES: We examined the associations among early-adolescent substance use, subsequent young-adult substance use disorders, and psychiatric disorders among a community sample of males. METHODS: Early-adolescent data were collected in classroom surveys (1990-1993), and young-adult data were collected in face-to face interviews (1998-2000). RESULTS: We found strong associations between early adolescent substance use and young-adult substance use disorders and psychiatric disorders. The magnitudes of these associations varied by racial/ethnic group and were strongest among African Americans and foreign-born Hispanics, who reported the lowest early-adolescent substance use. CONCLUSIONS: Early-adolescent substance use is most strongly associated with a later pattern of dysfunction among the racial/ethnic groups that reported the lowest levels of early use. The implications of our findings in the context of primary and secondary prevention are discussed. PMID- 15333323 TI - Usefulness of tobacco check boxes on death certificates: Texas, 1987-1998. AB - OBJECTIVES: We compared reports of deaths in which tobacco use was a contributing factor ("tobacco-associated deaths") before and after the addition to death certificates in Texas of a check-box question asking whether tobacco use contributed to an individual's death. METHODS: We examined Texas vital statistics files from 1987 to 1998. We calculated differences in percentages of reported tobacco-associated deaths (and 95% confidence intervals [CIs]) for the periods 1987 to 1992, before the addition of the check-box question, and 1993 to 1998, after the addition of the check-box. RESULTS: Reports of tobacco-associated deaths were significantly less frequent before addition of the check-box question (0.7%; 95% CI = 0.4%, 1.0%) than after addition of the question (13.9%; 95% CI = 13.0%, 14.7%). From 1993 to 1998, percentages of tobacco-associated deaths reported on the check-box question increased steadily. CONCLUSIONS: The addition of a tobacco-associated-death check box on Texas death certificates significantly increased reporting of tobacco use contributions to mortality. PMID- 15333324 TI - Long-term ethylene oxide exposure trends in US hospitals: relationship with OSHA regulatory and enforcement actions. AB - OBJECTIVES: We assessed long-term trends in ethylene oxide (EtO) worker exposures for the purposes of exposure surveillance and evaluation of the impacts of the Occupational Safety and Health Administration (OSHA) 1984 and 1988 EtO standards. METHODS: We obtained exposure data from a large commercial vendor and processor of EtO passive dosimeters. Personal samples (87,582 workshift [8-hr] and 46,097 short-term [15-min] samples) from 2265 US hospitals were analyzed for time trends from 1984 through 2001 and compared with OSHA enforcement data. RESULTS: Exposures declined steadily for the first several years after the OSHA standards were set. Workshift exposures continued to taper off and have remained low and constant through 2001. However, since 1996, the probability of exceeding the short-term excursion limit has increased. This trend coincides with a decline in enforcement of the EtO standard. CONCLUSIONS: Results indicate the need for renewed intervention efforts to preserve gains made following the passage and implementation of the 1984 and 1988 EtO standards. PMID- 15333325 TI - Hepatitis A virus immunity and seroconversion among contacts of acute hepatitis A patients in Amsterdam, 1996-2000: an evaluation of current prevention policy. AB - OBJECTIVES: We evaluated the hepatitis A virus (HAV) control policy (hygienic precautions and passive immunization with immune globulin) for "household contacts" (defined as all people who lived in the same house and who shared the same toilet with the patient, people who took care of an HAV-infected child, and sexual partners of the patient) of acute hepatitis A patients between 1996 and 2000. METHODS: We examined the characteristics and the serological outcomes of household contacts. All susceptible contacts were invited for retesting 6 weeks after they received immune globulin. RESULTS: Of 1242 contacts of 569 HAV patients, more than 50% (n = 672) were found to be HAV immune. Among the remaining contacts, 161 (28.2%) had a concurrent infection, and 86 of these individuals were symptomatic. The remaining 409 susceptible contacts received immune globulin, with 186 (45%) returning for retesting 6 weeks later (64 [34%] were infected, but only 12 had symptoms). CONCLUSIONS: Immune globulin does not protect all household contacts from HAV infection; however, it attenuates symptoms and effectively reduces further HAV transmission. PMID- 15333326 TI - Cigarette smoke induces senescence in alveolar epithelial cells. AB - Cellular senescence is a state of irreversible growth arrest induced either by telomere shortening (replicative senescence) or by telomere-independent signals (stress-induced senescence). The alveolar epithelium is often injured by a variety of inhaled toxins, including cigarette smoke (CS). In the present study, we investigated whether exposure to CS induces senescence of alveolar epithelial cells. In vitro experiments showed that exposure of A549 cells or normal human alveolar epithelial cells to sublethal concentrations of aqueous CS extracts induced cellular senescence. The senescence was characterized by a dose- and time dependent increase in senescence-associated beta-galactosidase activity, senescence-associated changes in cell morphology, an increase in cell size and lysosomal mass, accumulation of lipofuscin, overexpression of p21(CIP1/WAF1/Sdi1) protein, and irreversible growth arrest. In vivo experiments in Institute for Cancer Research mice showed that inhalation of CS for 2 wk induced increases in senescence-associated beta-galactosidase activity, lipofuscin accumulation, and p21(CIP1/WAF1/Sdi1) protein expression in alveolar epithelial cells. These results suggest that CS induces a phenotype that is indistinguishable from that of senescence in alveolar epithelial cells. The induction of cellular senescence by CS may contribute to impaired re-epithelialization, leading to CS-related chronic lung diseases. PMID- 15333327 TI - Cigarette smoke alters chromatin remodeling and induces proinflammatory genes in rat lungs. AB - Cigarette smoke-triggered inflammation is considered to play a central role in the development of chronic obstructive pulmonary disease by a mechanism that may involve enhanced proinflammatory gene transcription. Histone acetylation and deacetylation is a key regulator of the specificity and duration of gene transcription. Disruption in the nuclear histone acetylation:deacetylation balance (chromatin remodeling) may result in excessive transcription of specific proinflammatory genes in the lungs. In this study we show that cigarette smoke exposure results in an influx of inflammatory cells and chromatin modifications in rat lungs. This was associated with an increase in the active phosphorylated form of p38 mitogen-activated protein kinase concomitant with increased histone 3 phospho-acetylation, histone 4 acetylation, and increased DNA binding of the redox-sensitive transcription factor nuclear factor-kappaB, independent of inhibitory protein-kappaB degradation, and activator protein 1. We also observed decreased histone deacetylase 2 activity, which is due to protein modification by aldehydes and nitric oxide products present in cigarette smoke. Furthermore, we show that corticosteroid treatment has no effect on smoke-induced proinflammatory mediator release. These findings suggest a possible molecular mechanism by which cigarette smoke drives proinflammatory gene transcription and an inflammatory response in the lungs. PMID- 15333328 TI - Conditional overexpression of bioactive transforming growth factor-beta1 in neonatal mouse lung: a new model for bronchopulmonary dysplasia? AB - Research interest in bronchopulmonary dysplasia (BPD) has steadily increased, and numerous potential mediators have been implicated in the development of the disease. Among such mediators is transforming growth factor (TGF)-beta. Unfortunately, commonly utilized murine transgenic models are not optimal to investigate the effects of TGF-beta specifically during the 2-3 wk period of alveolar formation, the developmental stage that corresponds histologically to early alveolar development in humans, and the time frame during which BPD develops. In the current study, we utilized a triple-transgenic construct to overexpress bioactive TGF-beta1 in the neonatal mouse lung during the period of alveolar formation. Lungs were then examined by histologic, Western blot, and immunofluorescent methods. We found that overexpression of bioactive TGF-beta1 in neonatal mouse lungs resulted in structural changes that have been described in BPD. Included in those characteristics are abnormal alveolar structure, cellular composition, and vascular development. Our study indicates that TGF-beta1 overexpression in the neonatal mouse lung results in histologic alterations that have striking similarities to pathologic descriptions of BPD. We encourage the use of conditional transgenic models for the study of BPD, and hypothesize that the TGF-beta system is a central mediator for the histologic alterations described in association with the disease. PMID- 15333329 TI - Transforming growth factor-beta antagonizes alveolar type II cell proliferation induced by keratinocyte growth factor. AB - Keratinocyte growth factor (KGF) is a mitogen for rat type II cells and also stimulates differentiation in vitro. Administration of KGF also protects the lung from a variety of injuries and subsequent development of fibrosis. Because transforming growth factor (TGF)-beta has been shown to inhibit epithelial cell proliferation and surfactant protein gene expression in other systems and is thought to be a major effector in pulmonary fibrosis, we sought to determine if TGF-beta would antagonize the effects of KGF in primary cultures of alveolar type II cells. Type II cells were cultured on a matrix of type I collagen and Matrigel in the presence or absence of KGF and/or TGF-beta. KGF alone greatly stimulated proliferation and increased cyclin-dependent kinase (cdk) 2 kinase activity and Retinoblastoma susceptibility gene product (Rb) phosphorylation. Cyclin D1, cdk2, and cdc25A protein levels were increased, and p15(Ink4b) and p27(Kip1) protein levels were decreased. TGF-beta markedly inhibited alveolar epithelial cell proliferation induced by KGF. TGF-beta inhibited cdk2 enzyme activity and Rb phosphorylation and increased p15(Ink4b) protein levels. TGF-beta also inhibited differentiation induced by KGF as measured by secretion of surfactant protein-A into the apical media. In summary, TGF-beta inhibits the proliferative effect of KGF in vitro and may be a biologic antagonist of KGF. PMID- 15333330 TI - Acute allergen-induced airway remodeling in atopic asthma. AB - Studies in animals and in human atopic skin suggest that allergen challenge may activate acute tissue remodeling changes via transforming growth factor-beta pathways. We determined whether inhalational allergen challenge in subjects with mild asthma induces similar acute changes to the airway epithelial mesenchymal trophic unit (EMTU). Endobronchial mucosal biopsies obtained before and 24 h after challenge were examined by confocal microscopy for extracellular matrix deposition in the reticular basement membrane (RBM). Cells actively involved in extracellular matrix synthesis were identified as immunoreactive to heat shock protein 47, a chaperone of collagen synthesis. Interleukin-4/13 and transforming growth factor-beta-activated cells were identified by specific antibodies to phosphorylated (phospho-) signal transducer and activator of transcription 6 and phospho-Smad2, respectively. After allergen challenge, there was a significant increase in the number of heat shock protein 47-positive airway fibroblasts (P = 0.003) and in the thickness of tenascin in the RBM (P = 0.031). There were also increases in the number of phospho-Smad2+ epithelial cells (P = 0.04) and nuclear phospho-Smad2+ fibroblasts (P = 0.03), as well as phospho-signal transducer and activator of transcription 6+ epithelial cells (P = 0.03), after allergen challenge. Thus, allergen challenge in patients with mild asthma induces activation of epithelial cells and fibroblasts in the EMTU as well as increased tenascin deposition within the RBM. Airway remodeling in asthma may, in part, result from repeated acute activation of the EMTU by allergen exposure. PMID- 15333331 TI - Importance of tumor necrosis factor-alpha cleavage process in post transplantation lung injury in rats. AB - Tumor necrosis factor-alpha (TNF-alpha) has two forms with apparently different biological activities: a membrane-associated form and a soluble form. TNF-alpha converting enzyme (TACE) mediates a cleavage of membrane-associated TNF-alpha to induce its bioactive soluble form. We hypothesized that inhibition of TACE might prevent TNF-alpha-induced tissue injury while preserving the benefits of TNF alpha. In this study, we evaluated the role of TACE in acute inflammation using an inhibitor of the enzyme in a rat model of lung transplantation. Inbred Lewis rats underwent left lung isotransplantation, and the donor lungs were kept in Euro-Collins solution with or without the inhibitor. After 6 hours of ischemia, the left lung was transplanted into the recipient rat and reperfused for 4 hours. Inhibition of TACE significantly attenuated endothelial and alveolar septal damage, as assessed by radiolabeled albumin leakage after transplantation. The inhibition also attenuated neutrophil accumulation in the alveolar space and other histopathologic findings, including intercellular adhesion molecule-1 expression. In addition, significantly lower levels of monocyte chemotactic protein-1, cytokine-induced neutrophil chemoattractant-1, high mobility group box 1, and soluble epithelial cadherin and decreased neutrophil elastase activity were observed in bronchoalveolar lavage fluid from the rats treated with the inhibitor. We conclude that TACE mediates a critical step in the development of post-transplantation lung injury. PMID- 15333332 TI - Ventilation asymmetry after transplantation for emphysema: role of chest wall and mediastinum. AB - After single-lung transplantation for emphysema, the hyperinflated native lung and the graft have different extents and rates of inflation and emptying. This requires that breathing produces asymmetrical expansion of the chest wall, displacement of the mediastinum, or both. In a first study in four seated transplant recipients, we measured the volumes of the two hemithoraces with optoelectronic plethysmography. Functional residual capacity and total lung capacity were identical on the native and transplanted sides, and changes in chest wall volume during CO(2)-induced hyperpnea and FVC maneuvers were similar on both sides. Studies with computerized tomography in three of these patients and in four additional patients in supine posture indicated that the mediastinum was shifted toward the graft at functional residual capacity and total lung capacity. The mediastinum moved toward the native lung during tidal and full inspiration and toward the graft during tidal and forced expiration; additional studies with fluoroscopy showed qualitatively similar changes in upright posture. In summary, the two hemithoraces assume identical static volumes and show similar volume changes during CO(2)-induced hyperpnea and FVC maneuvers in patients with single-lung transplantation for emphysema; displacement of the mediastinum accommodates part, if not all, of the unequal lung volumes and asymmetrical ventilation. PMID- 15333333 TI - C-type natriuretic peptide ameliorates monocrotaline-induced pulmonary hypertension in rats. AB - C-type natriuretic peptide (CNP) has been shown to act as a local regulator of vascular tone and remodeling. We investigated whether CNP ameliorates monocrotaline (MCT)-induced pulmonary hypertension in rats. Rats received a continuous infusion of CNP or placebo. Significant pulmonary hypertension developed 3 weeks after MCT. However, infusion of CNP significantly attenuated the development of pulmonary hypertension and vascular remodeling. Neither systemic arterial pressure nor heart rate was altered. Interestingly, CNP enhanced Ki-67 expression, a marker for cell proliferation, in pulmonary endothelial cells and augmented lung tissue content of endothelial nitric oxide synthase. CNP significantly suppressed apoptosis of pulmonary endothelial cells, decreased the number of monocytes/macrophages, and inhibited expression of plasminogen activator inhibitor type 1, a marker for fibrinolysis impairment, in the lung. In addition, CNP significantly increased the survival rate in MCT rats. Finally, infusion of CNP after the establishment of pulmonary hypertension also had beneficial effects on hemodynamics and survival. In conclusion, infusion of CNP ameliorated MCT-induced pulmonary hypertension and improved survival. These beneficial effects may be mediated by regeneration of pulmonary endothelium, inhibition of endothelial cell apoptosis, and prevention of monocyte/macrophage infiltration and fibrinolysis impairment. PMID- 15333334 TI - Outcome bias. PMID- 15333335 TI - Increase in utilization of percutaneous renal artery interventions by medicare beneficiaries, 1996-2000. AB - OBJECTIVE: Our purpose was to review data from Medicare physician claims to ascertain differences in annual volumes of renal artery interventions versus surgery and differences in utilization of renal artery interventions among geographic regions according to physician provider type between 1996 and 2000. MATERIALS AND METHODS: We analyzed claims submitted to Medicare in 1996, 1998, and 2000 and extracted claims for renal artery angioplasty, stent placement, or bypass surgery. Analyses were performed for percutaneous renal artery interventions categorized by Centers for Medicare & Medicaid Services (CMS) geographic region and physician provider type. RESULTS: Between 1996 and 2000, the total volume of renal revascularization (surgical and percutaneous) increased 62%, from 13,380 to 21,660 procedures. The annual volume of renal artery surgery decreased 45% in 2000, compared with the volume in 1996. Annual volumes of renal artery angioplasty and stent placement increased 2.4-fold in 2000 compared with those in 1996. Most growth in percutaneous renal artery interventions is attributed to added provision by cardiologists, who increased their annual volume 3.9-fold. More than a threefold difference in rates of use of renal artery interventional procedures across CMS regions was found. In the Southeast region, the volume of renal artery interventions by cardiologists increased more than 15 fold. CONCLUSION: Among Medicare beneficiaries, the volume of percutaneous renal artery interventions is increasing rapidly, whereas the volume of renal artery surgery is declining. Most growth in percutaneous renal artery revascularization is attributed to increased performance by cardiologists; explosive growth in annual procedure volume by cardiologists occurred in some regions. Marked disparity in use among CMS regions was found. PMID- 15333336 TI - Incorporating ethics education into the radiology residency curriculum: a model. PMID- 15333337 TI - Ethics education in the radiology residency curriculum. PMID- 15333338 TI - Evaluation of patients referred for percutaneous ablation of renal tumors: importance of a preprocedural diagnosis. AB - OBJECTIVE: Percutaneous ablation of renal cell carcinoma has the potential to be as effective as partial nephrectomy. Because the entire tumor cannot be examined at pathology, diagnosis relies solely on imaging and percutaneous biopsy. Diagnoses of cancer have been rendered using imaging alone in some clinical trials of percutaneous ablation. If these trials inadvertently included benign masses, the efficacy of ablation was overestimated. Therefore, we sought to determine the prevalence of benign masses in a population of patients referred for percutaneous tumor ablation of presumed renal cell carcinoma. SUBJECTS AND METHODS: Twenty-seven patients were referred by urologists for MRI-guided cryotherapy of suspected renal cell carcinoma. Renal masses ranged from 1.0 to 4.6 cm (mean, 2.2 cm) in diameter. The CT, MRI, and percutaneous biopsy findings were tabulated and compared with surgical and imaging follow-up. RESULTS: Ten patients (37%) had a benign renal mass, including three angiomyolipomas, that had no evidence of fat on CT or MRI. Three masses were proven benign by biopsy, three by imaging, and four by a combination of biopsy and imaging. The masses in the remaining 17 patients were ablated. Biopsy revealed malignant cells in 10, suspicious cells in four, and atypical cells in two; one was nondiagnostic. CONCLUSION: A substantial percentage of patients referred for percutaneous ablation of renal tumors had benign masses. If CT or MRI alone cannot be used to diagnose a benign entity, patients should undergo a biopsy before the treatment session. PMID- 15333339 TI - Percutaneous sonographically guided radiofrequency ablation with artificial pleural effusion for hepatocellular carcinoma located under the diaphragm. AB - OBJECTIVE: Sonographically guided radiofrequency ablation is usually one of the most effective treatments for hepatocellular carcinoma. However, the localization of the tumor is a major limiting factor in the use of a sonographically guided procedure. In our experience, sonographic examination with artificial pleural effusion has improved the visualization of hepatocellular carcinomas under the diaphragm. We investigated the safety, benefits, and local efficacy of radiofrequency ablation with artificial pleural effusion. SUBJECTS AND METHODS: Twenty-five lesions in 23 patients were treated using radiofrequency ablation with artificial pleural effusion, for which 5% glucose solution was injected into the pleural cavity. RESULTS: Artificial pleural effusion allowed us to visualize the whole tumor on gray-scale sonography in 22 lesions that were not detectable or were poorly visible and to obtain a safer and easier puncture line in 14 lesions. In 23 (92%) of the 25 lesions, artificial pleural effusion was helpful in performing percutaneous radiofrequency ablation. Complete necrosis after radiofrequency ablation was obtained in 22 (88%) of the 25 lesions. During a mean (+/- SD) follow-up period of 10.6 +/- 6.5 months, local recurrence at the ablation site was diagnosed in only one (4.5%) of the 22 lesions. Mild cough in three patients and mild dyspnea in two patients were observed as adverse effects of artificial pleural effusion, but these effects were temporary. Oxygen saturation of the blood during artificial pleural effusion was slightly decreased. CONCLUSION: Radiofrequency ablation with artificial pleural effusion is a safe and beneficial treatment option that offers excellent local control through visualization of hepato-cellular carcinomas under the diaphragm. PMID- 15333340 TI - Hybrid treatment of acute massive pulmonary thromboembolism: mechanical fragmentation with a modified rotating pigtail catheter, local fibrinolytic therapy, and clot aspiration followed by systemic fibrinolytic therapy. AB - OBJECTIVE: We sought to evaluate the efficacy and safety of a hybrid treatment for acute massive pulmonary thromboembolism in patients with hemodynamic impairment by combining mechanical fragmentation, local thrombolysis, and clot aspiration. SUBJECTS AND METHODS: Within a period of 35 months, 25 patients with hemodynamic impairment (eight men and 17 women; age range, 35-77 years) were treated with mechanical thrombus fragmentation using a modified rotating pigtail catheter. After embolus fragmentation, all patients received an intrapulmonary injection of recombinant human-tissue plasminogen activator and then underwent manual clot aspiration with a large-lumen percutaneous transluminal coronary angioplasty guide catheter. RESULTS: All the patients survived, and their clinical status improved. Posttreatment angiography showed an improvement in pulmonary perfusion in all patients (mean Miller score before treatment, 22.2; after treatment, 13.6; p < 0.01). Mean pulmonary artery pressure decreased from 32.6 to 23.4 mm Hg (p < 0.01). Mean treatment time was 124.6 min. CONCLUSION: Hybrid treatment with mechanical fragmentation using a rotating pigtail catheter combined with local fibrinolysis and manual clot aspiration resulted in a rapid and safe improvement in the hemodynamic condition of patients with acute massive pulmonary thromboembolism. This hybrid treatment appears to be especially useful in patients at high risk for right ventricular failure and is a minimally invasive alternative to surgical embolectomy. PMID- 15333341 TI - Irrigation of the bile ducts with chilled saline during percutaneous radiofrequency ablation of a hepatic ocular melanoma metastasis. PMID- 15333342 TI - MRI-guided radiofrequency thermal ablation of normal lung tissue: in vivo study in a rabbit model. AB - OBJECTIVE: The purpose of this study was to assess the feasibility of MRI to guide and monitor radiofrequency ablation of normal pulmonary tissue in a rabbit model. MATERIALS AND METHODS: Percutaneous puncture and lung radiofrequency ablation were performed in six New Zealand white rabbits under MRI control using a 0.2-T open MRI scanner. Technical feasibility and complication detection were evaluated. The ablation zone appearance and size were assessed using MRI, CT, and gross pathology. Interclass correlation coefficients (ICCs) of the maximum short axis diameters of the lesions on gross pathology and the corresponding diameters as measured on each MRI pulse sequence and on CT scans were calculated. RESULTS: MRI guidance of percutaneous puncture and radiofrequency ablation of pulmonary tissue is feasible. A pneumothorax was detected and treated using MRI. In the specimen, the mean coagulation necrosis diameter was 9.8 mm. The T1-weighted spoiled gradient-echo fast low-angle shot images showed the highest ICC (0.81) for the thermal lesion diameter. CONCLUSION: Our results indicate that MRI guidance is feasible and useful for radiofrequency ablation of normal pulmonary tissue. PMID- 15333343 TI - Percutaneous radiofrequency ablation of lung tumors with expandable needle electrodes: tips from preliminary experience. PMID- 15333344 TI - Angio-seal device used for hemostasis in the descending aorta. PMID- 15333345 TI - Ankle and foot injuries: analysis of MDCT findings. AB - OBJECTIVE: The purpose of our study was to assess MDCT findings and the advantages of MDCT compared with radiography in patients referred to a level 1 trauma center for diagnostic evaluation of acute ankle and foot trauma. MATERIALS AND METHODS: During a period of 37 months, 388 patients underwent MDCT of the ankle and foot due to an acute trauma. Imaging studies (MDCT and radiography) were retrospectively reevaluated with respect to fracture location, fracture type, and mechanism of injury, and findings from the primary radiographs of the ankle and foot were compared with MDCT findings. RESULTS: Of the 388 patients, 344 (89%) had one or more fractures in the ankle or foot. A total of 517 fractures were found in all anatomic regions: ankle, calcaneus, talus, midfoot, and forefoot. The three most common occult fractures in the ankle not detected on primary radiography were isolated fractures of the posterior and medial malleolus and Tillaux fractures. The calcaneus was the most commonly fractured bone, and the sensitivity of radiography in the detection of calcaneal fractures was 87%. The sensitivity of radiography in the detection of talar fractures was 78%, whereas it was only 25-33% in the detection of midfoot fractures. A Lisfranc fracture-dislocation was not detected on primary radiography in five (24%) of 21 cases. The three main injury mechanisms were falling from a height (164 patients [48%]), a simple fall (68 patients [20%]), and a traffic accident (47 patients [14%]). CONCLUSION: In patients with injuries from high-energy polytrauma and in those with complex ankle and foot fractures, the sensitivity of radiography is only moderate to poor; in these cases, MDCT is recommended as the primary imaging technique. PMID- 15333346 TI - Tuberculous infection of the wrist: MRI features. AB - OBJECTIVE: The objective of our study was to define the MRI features of tuberculous infection of the wrist. MATERIALS AND METHODS: We present the MRI findings of eight patients with tuberculous infection of the wrist. Spin-echo T1 weighted, gradient-echo T2(*)-weighted, and fast spin-echo T2-weighted sequences were performed for all patients. Gadolinium-enhanced MR images were obtained in seven patients. All images were evaluated for the characteristics of tuberculous infection of the wrist, including the presence of synovial thickening around the joints and tendons, signal intensity of the thickened tenosynovium and synovium on the T2-weighted images, synovial fluid collection in the tendon sheath, small low-signal and nonenhanced foci in the synovial fluid, bone erosion, osteomyelitis, and encasement of the median nerve. RESULTS: The tuberculous infection involved the right (n = 6) and left (n = 2) wrists. All patients had synovial thickening around the flexor and extensor tendons with synovial fluid collection in the tendon sheath. The thickened tenosynovium and synovium revealed low signal intensity on T1-weighted images, intermediate to low signal intensity on T2- and T2(*)-weighted images, and enhancement on contrast-enhanced MR images. The synovial fluid showed intermediate to low signal on T1-weighted images and homogeneous or heterogeneous high signal intensity on T2- and T2(*)-weighted images. Multiple small foci of low signal intensity and nonenhancement scattered in the synovial fluid were present in seven patients. Bone erosion occurred in seven patients, osteomyelitis was seen in six patients, and encasement of the median nerve was found in three patients. CONCLUSION: Characteristic MRI findings of tuberculous infection of the wrist include synovial thickening around the flexor and extensor tendons and synovial fluid collection that contains small low signal and nonenhanced foci in the tendon sheath. Bone erosion, osteomyelitis, and median nerve encasement are also frequently present. These characteristic manifestations are helpful in diagnosing this disease entity. PMID- 15333347 TI - Extraaxial neurofibromas versus neurilemmomas: discrimination with MRI. AB - OBJECTIVE: The purpose of our study was to evaluate whether MRI can discriminate between extraaxial neurofibromas and neurilemmomas. MATERIALS AND METHODS: MR images of 52 patients with a pathologically proven extraaxial neurofibroma or neurilemmoma were retrospectively reviewed by observers who were unaware of the surgical results, regarding the presence or absence of individual imaging criteria. MRI findings in 12 patients with a localized neurofibroma and 40 patients with a neurilemmoma were compared using the chi-square test or Fisher's exact test. RESULTS: MRI findings suggestive of neurofibroma (p < 0.05) were a target sign on T2-weighted images (58% in neurofibromas vs 15% in neurilemmomas), central enhancement (75% vs 8%), and a combination of both findings (63% vs 3%). MRI findings suggestive of a neurilemmoma (p < 0.05) were a fascicular appearance on T2-weighted images (25% vs 63%), a thin hyperintense rim on T2-weighted images (8% vs 58%), a combination of both findings (8% vs 48%), and diffuse enhancement (13% vs 67%). No significant difference was seen between neurofibromas and neurilemmomas for a centrally entering and exiting nerve (42% in neurofibromas vs 23% in neurilemmomas), a peripherally entering and exiting nerve (58% vs 77%), a cystic area (38% vs 64%), a low-signal margin (100% vs 100%), peripheral enhancement (13% vs 26%), or a target sign on contrast-enhanced images (11% vs 31%). CONCLUSION: MRI shows features helpful for differentiating extraaxial neurofibromas from neurilemmomas; however, no single finding or combination of findings allows definitive differentiation. PMID- 15333348 TI - Progressive systemic sclerosis with massive paraspinal soft-tissue calcinosis. PMID- 15333349 TI - Asymptomatic tibial stress reactions: MRI detection and clinical follow-up in distance runners. AB - OBJECTIVE: The purpose of this study was twofold: to determine if asymptomatic elite distance runners exhibit stress reactions of the tibia on MR images and to determine if the presence of bone stress lesions predicts later development of symptomatic tibial stress injuries. CONCLUSION: Signs of a tibial stress reaction were found on MRI in 43% of the 21 asymptomatic college distance runners in this study. The presence of these changes was not found to be a predictor of future tibial stress reactions or stress fractures. Our findings underscore the importance of correlating MRI findings with clinical findings before making therapeutic decisions. PMID- 15333350 TI - Limited diagnostic agreement of quantitative sonography of the radius and phalanges with dual-energy x-ray absorptiometry of the spine, femur, and radius for diagnosis of osteoporosis. AB - OBJECTIVE: The aim of our study was to evaluate the diagnostic agreement of quantitative sonography of the radius and proximal phalanx and dual-energy X-ray absorptiometry (DXA) of the radius, lumbar spine, and femoral neck for the detection of osteoporosis. MATERIALS AND METHODS: In 95 women (mean age, 53 +/- 13 years) and 26 men (mean age, 53 +/- 13 years), DXA measurements of the lumbar spine (posterior-anterior, L1-L4) and the femoral neck, as well as quantitative sonography of the radius and proximal phalanx of the third finger were obtained. The percentage of patients below a given threshold was calculated for each imaging technique. A T score of less than -2.5 indicated presence of osteoporosis. Diagnostic agreement in identifying individuals with osteoporosis was assessed using kappa scores. RESULTS: Between 14% and 22% of the patients were classified as osteoporotic after DXA of the various regions of interest of the radius, 31% after DXA of the spine, 43% after DXA of the femoral neck, 32% after quantitative sonography of the distal radius, and 34% after quantitative sonography of the phalanx of the third finger. Correlation coefficients between T values for quantitative sonography and those for DXA varied between not significant and 0.54 at the different regions. Kappa analysis showed the diagnostic agreement among quantitative sonography and DXA to be fair to moderate (kappa = 0.38-0.48). The highest agreement was between quantitative sonography of the proximal phalanx of the third finger and DXA of the total radius (kappa 0.48; p < 0.05). CONCLUSION: Considerable diagnostic disagreement exists between quantitative sonography and DXA of the forearm, as is true for most quantitative techniques in the assessment of skeletal status. The lack of correlation makes quantitative sonography impractical for routine diagnostic use but might characterize different parameters related to bone quality. PMID- 15333351 TI - Effect of granulocyte-stimulating factors on marrow of adult patients with musculoskeletal malignancies: incidence and MRI findings. AB - OBJECTIVE: Our goal was to determine the incidence and pattern of red marrow reconversion on MRI of adults receiving granulocyte-stimulating factors as part of their chemotherapy regimen for primary musculoskeletal neoplasms and correlate the changes with WBC counts. MATERIALS AND METHODS: Twenty-five adults with soft tissue sarcomas (n = 15) or primary bone tumors (n = 10) who underwent chemotherapy that included granulocyte-stimulating factors formed the study group. Two radiologists retrospectively evaluated the MRI studies by consensus before and after therapy to determine the presence of changes consistent with red marrow reconversion. Changes were categorized by appearance on T1- and T2 weighted images for location, pattern, and extent of marrow involvement. WBC counts at the time of MRI were recorded. Records were examined for evidence of marrow-infiltrating tumors or metastases. RESULTS: Ten (40%) of the 25 patients underwent bone marrow changes consistent with red marrow reconversion. In seven (70%) of 10 patients, the changes were diffuse in visualized bones and simulated diffuse marrow-infiltrative tumor. In three (30%) of 10 patients, the changes were focal, simulating metastases. T2 signal prolongation was identified in the marrow reconversion in nine patients (90%), although all had shortened T1 signal. Four patients (40%) had elevated WBC counts at the time of the MRI after therapy, but all had shortened T1 signal. Four patients (40%) had elevated WBC counts at the time of the MRI after therapy, five (50%) had normal counts, and one (10%) had a below-normal count. No osseous metastases or marrow-infiltrating tumors were found during follow-up. CONCLUSION: Forty percent of patients showed marrow changes mimicking tumor on MRI that were attributable to red marrow reconversion, which correlated moderately with WBC response. PMID- 15333352 TI - Evaluation of combined transmission and emission tomography for classification of skeletal lesions. AB - OBJECTIVE: This study assessed the benefit of transmission emission tomography (TET) for classification of skeletal lesions in patients with known malignant disease. SUBJECTS AND METHODS: The TET technology combines acquisition of SPECT and CT data using the same imaging device, thus allowing perfect overlay of anatomic and functional images. We performed TET in 47 patients with tumors who had a total of 104 focal lesions found on bone scintigraphy. Technetium-99m diphosphonate was used as the radiopharmaceutical in all patients. Findings of bone scintigraphy (planar and SPECT), SPECT + CT or radiography, and TET were compared with regard to the precise location and nature (benign vs malignant) of each lesion. Validation was achieved by radiologic follow-up on CT, MRI, or radiography, especially for the extremities, and using biopsy results in five patients. RESULTS: TET could classify 88 (85%) of 104 lesions compared with 37 (36%) of 104 on SPECT. When we counted inconclusive studies as positive for cancer, discrepant findings between SPECT and TET were obtained in 39 lesions. In 38 (97%) of these, TET was correct. Sensitivity for cancer detection was 98% for TET and 94% for SPECT (p = 0.63), and specificity was 81% for TET and 19% for SPECT (p < 0.0001). The highest diagnostic gain was in the spine, thoracic cage, skull, and pelvis. Small osteolytic lesions were missed because of the limited resolution of transmission images. SPECT + CT or radiography and TET were discordant in nine of 104 lesions. TET was false-negative in one lesion and false positive in another, and SPECT + CT or radiography was false-positive in seven lesions. As a result, sensitivities of TET and SPECT + CT or radiography were nearly the same, but the specificity of TET was significantly higher (p = 0.015). CONCLUSION: TET improves the accuracy of bone scintigraphy by correctly classifying equivocal lesions, especially by identifying benign abnormalities in the axial skeleton and thus increasing the specificity of positive findings. PMID- 15333353 TI - Portable abdominal CT: analysis of quality and clinical impact in more than 100 consecutive cases. AB - OBJECTIVE: Our aim was to determine portable abdominal CT image quality and clinical content in a consecutive series of scans obtained in ICUs at a single center. MATERIALS AND METHODS: All helical portable abdominal CT scans obtained between June 1999 and December 2000 were reviewed by two observers. Image quality was assessed for feature detection and overall quality compared with the patients' contemporaneous stationary CT scans. Hospital records were used to determine patient demographics, scanning indications, and clinical impact and to verify portable CT findings when possible. RESULTS: One hundred twenty-two helical portable CT scans (47 contrast-enhanced) and 41 contemporaneous stationary CT scans in 107 patients were included. IV contrast material improved portable CT scan quality, but quality scores for portable CT scans were consistently lower than those for stationary CT scans, both with and without contrast material. Thirty-three conditions suspected before scanning were supported by findings on portable CT scans, which detected evidence of infection in 18 and hemorrhage in 16 cases and motivated seven laparotomies and six percutaneous drainage procedures. Thirty-three portable CT scans (27%) contributed to a change in patient treatment. Results of surgery or autopsy confirmed portable CT findings in 12 of 17 cases. CONCLUSION: Although image quality is inferior to conventional stationary CT, portable abdominal CT provides important diagnostic information without requiring patient transport outside the ICU. Radiologists should avoid overconfident interpretation of portable CT scans. PMID- 15333354 TI - MRI for clinically suspected appendicitis during pregnancy. AB - OBJECTIVE: The purpose of this study was to evaluate whether MRI can be used to accurately diagnose or exclude appendicitis in pregnant patients with clinically suspected appendicitis. CONCLUSION: Our results suggest that MRI is helpful in the examination and diagnosis of acute appendicitis in pregnant patients. MRI may therefore be a good alternative to CT in pregnant patients for whom sonographic findings are nondiagnostic. PMID- 15333356 TI - Abdominal wall hernias: MDCT findings. PMID- 15333355 TI - Parallel imaging and diffusion tensor imaging for diffusion-weighted MRI of the liver: preliminary experience in healthy volunteers. AB - OBJECTIVE: Our aim was to determine whether parallel imaging and diffusion tensor imaging affect the measurement of apparent diffusion coefficient (ADC) during diffusion-weighted MRI of the liver in healthy volunteers. SUBJECTS AND METHODS: We performed breath-hold single-shot echo-planar diffusion-weighted MRI of the liver in 10 healthy volunteers using conventional diffusion, conventional diffusion with parallel imaging, and diffusion tensor with parallel imaging sequences. TE values for the three sequences were 83, 74, and 63, respectively. Liver signal intensity was measured on all sequences and normalized to the SD of the measurement. Hepatic ADC was calculated by acquiring all sequences with b values of 0 and 500 sec/mm(2). RESULTS: The normalized liver signal intensity was higher on diffusion tensor with parallel imaging and conventional diffusion with parallel imaging than on conventional diffusion without parallel imaging for a b value of 500 sec/mm(2) (13.0 and 10.1 vs 9.1, respectively; p < 0.03) and for a b value of 0 sec/mm(2) (9.0 and 7.6 vs 6.9, respectively; without reaching a significant difference, p = 0.12). Hepatic ADC was not significantly different between sequences (p = 0.16). CONCLUSION: Higher signal intensity can be obtained when using parallel imaging and diffusion tensor imaging during diffusion weighted MRI of the liver without compromising hepatic ADC measurement. PMID- 15333357 TI - Intussusception in adults: from stomach to rectum. PMID- 15333358 TI - Multiarterial phase dynamic MRI of small early enhancing hepatic lesions in cirrhosis or chronic hepatitis: differentiating between hypervascular hepatocellular carcinomas and pseudolesions. AB - OBJECTIVE: The purpose of this study was to assess the value of multiarterial phase contrast-enhanced dynamic MRI of the whole liver obtained during a single breath-hold for small early enhancing hepatic lesions in patients with cirrhosis or chronic hepatitis, emphasizing the distinction between hypervascular hepatocellular carcinomas and pseudolesions. MATERIALS AND METHODS: The study population included 40 patients with cirrhosis or chronic hepatitis who had small early enhancing hepatic lesions (a total of 70 lesions: 40 hepatocellular carcinomas, 30 pseudolesions). All patients underwent multiarterial phase contrast-enhanced dynamic MRI (six phases) of the whole liver during a single breath-hold. RESULTS: Twenty-one (53%) of 40 hypervascular hepatocellular carcinomas showed rapid central washout after early enhancement of the lesion as well as peritumoral coronal enhancement, but these findings were not observed in any hypervascular pseudolesions (p < 0.001). In 19 hepatocellular carcinomas without rapid central washout, early enhancement of the lesion appeared at the second, third, or fourth phase (mean, 2.5 phases). In eight of these 19 hepatocellular carcinomas, lesion enhancement disappeared by the sixth phase. Conversely, in 30 hypervascular pseudolesions, early enhancement of the lesion appeared at the second, third, fourth, or fifth phase (mean, 3.0 phases). In 28 of these 30 pseudolesions, lesion enhancement continued until the sixth phase. CONCLUSION: Rapid central washout after the early enhancement of the lesion and coronal enhancement surrounding the lesion are highly specific and diagnostic findings of small hypervascular hepatocellular carcinomas if present at multiarterial phase contrast-enhanced dynamic MRI. Hypervascular pseudolesions tend to show prolonged enhancement during the arterial phase compared with hypervascular hepatocellular carcinomas. PMID- 15333359 TI - Three-dimensional assessment of MRI-guided percutaneous cryotherapy of liver metastases. AB - OBJECTIVE: We report our initial investigation of the use of a 3D method for assessing percutaneous tumor ablations. We hypothesized that these 3D techniques could be used to assess the technical success of ablations and that 3D metrics would be predictive of treatment response. CONCLUSION: Three-dimensional assessment of percutaneous tumor ablations provides a quantitative evaluation of the technical success of the procedure. Three-dimensional computer-based techniques can both quantify coverage of a tumor and create a virtual ablation margin for percutaneous procedures, akin to a surgical margin. Although results are preliminary, 3D metrics were useful in predicting treatment response. PMID- 15333360 TI - Dynamic contrast-enhanced MRI analysis of perfusion changes in advanced hepatocellular carcinoma treated with an antiangiogenic agent: a preliminary study. AB - OBJECTIVE: To evaluate the perfusion changes in advanced hepatocellular carcinoma (HCC) treated with the antiangiogenic agent thalidomide, we used dynamic contrast enhanced MRI. SUBJECTS AND METHODS: Dynamic contrast-enhanced MRI was performed before and during thalidomide treatment in seven patients with advanced unresectable HCC that had failed to respond to prior local therapy. A turbo fast low-angle shot sequence was performed in a 1.5-T MR scanner. An operator-defined region of interest was placed in the maximal enhancement region of the tumor site and adjacent tumor-free parenchyma of all patients. A time-intensity curve was plotted and analyzed. The peak enhancement in the first-pass study, the maximal enhancement, and the initial enhancement slope percentage in the first-pass study of the tumor and parenchyma were measured. The changes in these three perfusion parameters were estimated and correlated with clinical outcomes. The seven patients were categorized into two groups on the basis of their clinical outcomes: group A patients were those who had progressive disease, whereas group B patients were those who had stable disease or partial response. RESULTS: Four of the seven patients were classified as group A, and the other three were classified as group B patients. When comparing the MRI parameters for the tumors before and during treatment in group A and group B patients, we found a statistically significant difference for the peak enhancement in the first-pass study, the maximal enhancement, and the enhancement slope percentage in the first pass study. When comparing the parenchymal parameters, we found a statistically significant difference in the maximal enhancement and borderline significance in the peak enhancement in the first-pass study (p = 0.057) between group A and group B patients. CONCLUSION: The dynamic MRI parameters showed significant differences between two groups of patients with different clinical outcomes. PMID- 15333361 TI - Hepatic pseudotumor due to nodular fatty sparing: the diagnostic role of opposed phase MRI. AB - OBJECTIVE: We sought to describe the role of opposed-phase MRI in the diagnosis of nodular fatty sparing in the liver. CONCLUSION: Nodular fatty sparing of the liver displays characteristic findings on opposed-phase MRI. Because the lesion may otherwise mimic a hepatic tumor, knowledge of these findings may prevent unnecessary biopsy or inappropriate treatment. PMID- 15333362 TI - The intradecidual sign: is it reliable for diagnosis of early intrauterine pregnancy? AB - OBJECTIVE: Our aim was to determine the accuracy of the intradecidual sign for the diagnosis of intrauterine pregnancy and the exclusion of ectopic pregnancy. CONCLUSION: The intradecidual sign reliably excludes the presence of an ectopic pregnancy. The sensitivity for diagnosis of an intrauterine pregnancy increases when human chorionic gonadotropin levels are equal to or greater than 2,000 mIU/mL or the mean sac diameter is equal to or greater than 3 mm. It is of utmost importance to visualize this sign on multiple views with an unchanging appearance. PMID- 15333363 TI - MRI detection of uterine necrosis after uterine artery embolization for fibroids. PMID- 15333364 TI - Sonography of the abnormal fallopian tube. PMID- 15333365 TI - Atypical CT and MRI manifestations of mature ovarian cystic teratomas. PMID- 15333366 TI - Trends in the use of CT and radiography in the evaluation of facial trauma, 1992 2002: implications for current costs. AB - OBJECTIVE: CT has replaced conventional radiography of the face in many trauma centers. Concern exists that increased costs are associated with increased use of CT. Our goal was to compare the amount of CT and radiography performed for facial trauma at a level 1 trauma center in 1992 and in 2002 and to determine hospital costs for the imaging of these patients. MATERIALS AND METHODS: The changes in volume and types of facial imaging examinations were determined comparing 1992 and 2002. Hospital costs of different imaging examinations were determined for 2002. Current costs of imaging facial trauma were compared with what 2002 costs would have been if the practice pattern in 1992 had continued. RESULTS: In 1992, 890 patients were evaluated for facial trauma. Six hundred seventy-one had only radiography, 153 only CT of the face, and 66 both CT and radiography. In 2002, 828 patients were evaluated. Five hundred eighty-four patients had only CT of the face; 228, only radiography; and 16, both CT and radiography. The number of facial imaging examinations per patient in 1992 and 2002 was 1.23 and 1.03, respectively. The 2002 hospital cost of a facial CT examination was $121 and of a facial radiography series was $154. Using CT instead of radiography for evaluating facial injury resulted in an overall cost savings of 22% per patient in 2002. CONCLUSION: The availability of CT has not resulted in increased use of facial imaging. The increased use of CT from 1992 to 2002 results in decreased current costs for the hospital. PMID- 15333367 TI - Facial reconstruction of a wrapped Egyptian mummy using MDCT. AB - OBJECTIVE: Facial reconstruction of mummies and corpses in general is important in anthropological, medical, and forensic studies. The purpose of our study was to evaluate the role of MDCT examination for 3D facial reconstruction and report the results of multidisciplinary work performed by radiologists, anthropologists, and forensic police in reconstructing the possible physiognomy of an ancient Egyptian mummy. MATERIALS AND METHODS: Three-dimensional MDCT data were obtained from a well-preserved, completely wrapped Egyptian mummy from the collection of the Egyptian Museum in Torino, Italy, which dated from the XXII or XXIII dynasty (945-715 BC). Data were used as a model for the rapid prototyping stereolithographic technique, a method that allows the creation of a 3D model with digital data using synthetic materials such as a resin or nylon. RESULTS: The physical creation of the face was accomplished with boosting techniques performed by the progressive layering of plasticine on the nylon model according to the anthropometric data, the conditions of the soft residual dehydrated tissues, and the most accepted scientific and anthropological criteria. CT is the only noninvasive method for obtaining fundamental data for 3D reconstructions of the skull and the body, especially with wrapped mummies. CONCLUSION: Our multidisciplinary cooperative study produced a model of the face of an individual who lived nearly 3,000 years ago, which would not previously have been possible unless we unwrapped, destroyed, and altered the conservation of the bandages and the mummy. PMID- 15333368 TI - Correlation between the amplitude of cortical activation and reaction time: a functional MRI study. AB - OBJECTIVE: We sought to examine the correlation between reaction time and the amplitude of cortical activation during the performance of a visuomotor response time task in a functional MRI (fMRI) experiment. We hypothesized that the fMRI blood oxygenation level-dependent (BOLD) amplitude may have a negative correlation with a subject's reaction time: the lower the amplitude within the cortical areas along the visuomotor pathway, the slower the response. A larger amplitude of the fMRI signal would reflect faster response times. SUBJECTS AND METHODS: During a single-event fMRI experiment, the reaction times (in milliseconds) of 32 right-handed subjects responding to a visual cue were recorded. Analysis of the single-event paradigm using Statistical Parametric Mapping (SPM99) was performed, activation maps were produced for each subject, and then a random effects group analysis was performed. The maximum amplitudes of cortical activation (percent signal change) in four activated cortical regions were estimated and tabulated. The regions of interest included were the right and left occipital visual cortices, the supplementary motor area, and the left sensorimotor area. Simple and multiple regressions were performed between the mean reaction times of the subjects and the BOLD amplitudes in each region of interest and for the composite region of interest. RESULTS: The results showed significant negative associations between the reaction times and maximum amplitudes in the right occipital, left occipital, and left sensorimotor area cortical regions (p < 0.05). However, no significant association was found between reaction times and the amplitude within the supplementary motor area. When the effects of age and sex on these associations were analyzed, we found that age had an impact on the results for individual regions of interest in the left occipital and left sensorimotor areas, but the composite amplitude of activation remained significantly correlated with reaction times. CONCLUSION: The degree of signal change in BOLD fMRI response of the right occipital, left occipital, and left sensorimotor areas reflects the speed of performance during the visuomotor response time task by the subject. Thus, the amplitude of activation can be used as one parameter to assess change in function. PMID- 15333369 TI - MDCT of the left atrium and pulmonary veins in planning radiofrequency ablation for atrial fibrillation: a how-to guide. PMID- 15333370 TI - Safety and hemodynamic effects of pulmonary angiography in patients with pulmonary hypertension: 10-year single-center experience. AB - OBJECTIVE: We sought to examine the incidence of complications and change in pulmonary artery pressure in patients with pulmonary hypertension who were undergoing pulmonary angiography. MATERIALS AND METHODS: A retrospective review was performed for all patients who underwent pulmonary angiography over a 10-year period at a single institution. Patients with moderate pulmonary hypertension (pulmonary artery pressure, 30-59 mm Hg) and severe pulmonary hypertension (pulmonary artery pressure, >/= 60 mm Hg) served as the study population. Demographic data, clinical indication, pre- and postcontrast pulmonary artery pressure measurements, type of pulmonary hypertension, contrast agent volume, complications, and American Society of Anesthesiologists (ASA) classification were recorded for all patients and compared. RESULTS: Two hundred two of 612 patients who underwent pulmonary angiography had pulmonary hypertension. Moderate pulmonary hypertension was present in 155 patients (77%) and severe pulmonary hypertension, in 47 patients (23%). Three (2.0%) of four complications were fatal. The complication rate was higher in patients with severe pulmonary hypertension compared with patients with moderate pulmonary hypertension but not statistically significant (6.3% vs 0.6%, p = 0.63). Patients with complications had a higher mean ASA score than those without complications (4.0 vs 3.0, p = 0.03). Patients with lung transplants had the greatest increase in pulmonary artery pressure after pulmonary angiography compared with all other clinical indications (16.75 +/- 12.97 mm Hg vs 5.46 +/- 6.86 mm Hg, p = 0.003). CONCLUSION: The complication rate of pulmonary angiography in patients with pulmonary hypertension is low. However, in severely ill patients with acute pulmonary hypertension, pulmonary angiography should be undertaken with extreme caution. PMID- 15333371 TI - Evaluation of classic 2D time-of-flight MR angiography in the depiction of severe carotid stenosis. AB - OBJECTIVE: The purpose of this study is to determine the sensitivity, specificity, and clinical utility of classic 2D time-of-flight MR angiography (acquired with derated gradients) as an aid to predicting severe carotid stenosis. SUBJECTS AND METHODS: Our study population was composed of 68 patients, yielding 133 carotid bifurcations for analysis. A 2D time-of-flight MR angiography pulse sequence was modified to provide greater sensitivity for carotid stenosis, which resulted in visualization of a carotid stenosis with a 70% or greater diameter as a signal void. Contrast-enhanced MR angiography was performed with the elliptical centric view order. Multiple overlapping thin-slab acquisition (MOTSA) MR angiography was performed in select patients. Digital subtraction angiography was performed in 51 patients, and the findings were used as the gold standard. In the remaining patients, findings on carotid duplex Doppler sonography and at surgery and clinical follow-up were used as the gold standard. RESULTS: In 51 patients for whom a digital subtraction angiogram was available, we found that the sensitivity of classic 2D time-of-flight MR angiography for prediction of carotid stenosis with a 70% or greater diameter was 94%, and the specificity of the technique was 97%. In three patients with severe carotid stenosis, the stenoses that appeared as signal voids on the classic 2D time-of-flight MR angiography were underestimated on contrast-enhanced MR angiography. Severe stenosis was confirmed by subsequent digital subtraction angiography, surgical results, or both. Discrepancies between findings on MOTSA MR angiography and contrast-enhanced MR angiography were resolved with classic 2D time-of-flight MR angiography. Classic 2D time-of-flight MR angiography increased diagnostic confidence of a severe stenosis in three patients with focal internal carotid artery stenosis. CONCLUSION: Classic 2D time-of-flight MR angiography has a high sensitivity and specificity for predicting carotid bifurcation stenosis of 70% or greater diameter. These probability measures allowed the detection of three significant stenoses that would have been missed on contrast-enhanced MR angiography and provided greater diagnostic confidence than contrast-enhanced or MOTSA MR angiography alone. PMID- 15333372 TI - Potential impact of pediatric MR urography on the imaging algorithm in patients with a functional single kidney. AB - OBJECTIVE: The purpose of this study was to evaluate the potential of MR urography in the assessment of children with a suspected "functional single kidney." SUBJECTS AND METHODS: Sixty patients (age range, 2.7 weeks to 15.7 years) who had been referred for assessment of a suspected functional single kidney underwent MR urography in addition to detailed sonography of the urinary tract and the currently indicated standard imaging. The results of the conventional imaging ((99m)Tc-dimer captosuccinic acid scintigraphy, voiding cystourethrography, and genitography) were compared with the results of sonography and MR urography; surgical findings served as the gold standard if available. RESULTS: Twenty-six patients had a single kidney. The other diagnoses were six contralateral multicystic dysplastic kidneys, two normal ectopic kidneys, one crossfused double system, and 25 ectopic or dysplastic renal buds. Scintigraphy detected all normal kidneys, two ectopic kidneys, and two dysplastic renal buds. Detailed sonography missed two ectopic kidneys and two orthotopic dysplastic renal buds, but one additional renal bud that could not be confirmed on other imaging techniques (accuracy, 91.7%; sensitivity, 88.2%; specificity, 96.2%) was suspected. MR urography results were correct in all patients and verified in all 13 with surgical correlation. CONCLUSION: MR urography allows a reliable assessment of renal and ureteral anatomy and of dysplastic or ectopic renal buds, even in non- or poorly functioning systems. MR urography therefore has the potential to replace the currently used excretory urography and scintigraphy. In patients with a suspected functional single kidney a detailed sonographic study and MR urography should be considered the diagnostic algorithm of choice. PMID- 15333373 TI - Sonographic measurement of the abdominal esophagus length in infancy: a diagnostic tool for gastroesophageal reflux. AB - OBJECTIVE: Our aim was to provide exact sonographic measurements of the abdominal esophagus length in neonates and infants with and without gastroesophageal reflux (GER) and to investigate its diagnostic value. GER severity and hiatal hernia presence were also evaluated and correlated with esophageal length. MATERIALS AND METHODS: This retrospective case-control study comprised 258 neonates and infants (150 without reflux and 108 with reflux). There were 50 children without reflux in each of three age categories: less than 1 month, 1-6 months, and 6-12 months. Of the children with reflux, 42 were less than 1 month old; 34, 1-6 months; and 32, 6-12 months. The abdominal esophagus was measured from its entrance into the diaphragm to the base of gastric folds in fed infants. The number of refluxes during a 10-min period were recorded; GER was categorized as mild, one to three refluxes; moderate, three to six refluxes; and severe, more than six refluxes. Presence of hiatal hernia was recorded. RESULTS: Neonates and infants with reflux had a significantly shorter abdominal esophagus than subjects without reflux: the mean difference in neonates, 4.8 mm; 1-6 months, 4.5 mm; 6-12 months, 3.4 mm. Children with severe reflux had a shorter esophagus compared with those with mild and moderate reflux only in the neonate group. In contrast, children with reflux associated with hiatal hernia had a significantly shorter esophagus compared with children with mild reflux in all three age groups. Sonographic sensitivity was 94%. CONCLUSION: Sonographic measurement of the abdominal esophagus length is highly diagnostic for GER in neonates and infants. In neonates, it can also indicate GER severity. Hiatal hernia is associated with a significantly shorter abdominal esophagus. PMID- 15333374 TI - Low-dose chest CT: optimizing radiation protection for patients. AB - OBJECTIVE: The aim of our study was to evaluate CT scanning protocols to determine how best to minimize patient exposure to ionizing radiation while maintaining sufficient image quality to detect pulmonary diseases. SUBJECT AND METHODS: The CT dose index (CTDI) was determined by scanning an acrylic phantom at various tube current-time products (7.5-115 mAs). Image quality was evaluated by comparing the homogeneity and noise level of CT scans obtained in the acrylic phantom with those obtained in a water-equivalent phantom. The CT scans obtained at various milliampere-second settings in patients with nodules or diffuse opacifications were assessed. The relationships between the CTDI and the image quality of the CT scans (noise level and artifacts) were established. RESULTS: The reduction of a conventional tube current-time product (115 mAs) by 65%, 78%, or 93.5% can decrease the CTDI by 60%, 70%, or 85%, respectively. In correlating the image quality of each CT scan to the milliampere-second settings used to obtain it, we found that homogeneity decreased as milliampere-second settings decreased, whereas the noise level increased as milliampere-second settings decreased. For both 8- and 3-mm slice thicknesses, the homogeneity of CT scans acquired at 7.5 mAs or greater was within the acceptable range (< 4 H). However, the noise level of CT scans remained within the acceptable range (< 0.35%) when 25 mAs or greater was used. The evaluation of the image quality of the patients' CT scans indicated no statistical significance in image quality rating between the scans obtained at 25 mAs and those obtained at 115 mAs (p > 0.01). CONCLUSION: Low-dose (i.e., 40 or 25 mAs) helical chest CT produced satisfactory image quality and reduced the CTDI, thereby maximally protecting patients from radiation exposure. PMID- 15333375 TI - Asthma and associated conditions: high-resolution CT and pathologic findings. PMID- 15333376 TI - Safety and MRI artifact evaluation at 1.5 T of metallic mounting sheath of a marking clip inadvertently deployed at stereotactic biopsy. AB - OBJECTIVE: The purpose of our study was to assess the MRI risk factors for the patient after an unusual case of inadvertent deployment of the mounting sheath of a metallic clip used at stereotactic breast biopsy. MATERIALS AND METHODS: We evaluated the materials for ferromagnetic properties, heating, and artifacts. RESULTS: Our analysis showed significant deflection and prominent susceptibility artifacts of the sheath at 1.5 T, although the clip itself showed no deflection and only minimal artifact. Our study shows that a device and its delivery apparatus may have significantly different ferromagnetic properties. CONCLUSION: In case of inadvertent deployment of the sheath, a delay of 6-8 weeks before MRI is recommended as a conservative approach to ensure tissue ingrowth and to minimize the chance of harm to the patient. In the case of metallic clips used for breast biopsy, caution is warranted when a portion of the device unintended for placement is introduced into the breast. PMID- 15333377 TI - Tattoo pigment mimicking axillary lymph node calcifications on mammography. PMID- 15333378 TI - Bilateral extraadrenal perirenal myelolipomas: an imaging challenge. PMID- 15333379 TI - Giant mature cystic teratoma of the adrenal region. PMID- 15333380 TI - Normal anatomy and disease processes of the pancreatoduodenal groove: imaging features. PMID- 15333381 TI - Digital image editing using PowerPoint: part 1, introduction to the image manipulation commands. AB - OBJECTIVE: This article, the first in a two-part series, reviews the manipulation commands of PowerPoint and briefly describes their advantages and limitations. CONCLUSION: The 2002 version of PowerPoint has many useful image-manipulation features. The second article will discuss some practical applications of these features. PMID- 15333382 TI - Digital image editing using PowerPoint: part 2, practical applications of the image-manipulation commands. AB - OBJECTIVE: This article, the second of two parts, suggests some practical applications for image manipulation using the 2002 version of PowerPoint. CONCLUSION: Applications include enhancing previously manipulated images and manipulating unedited images. The user can develop his or her preferred techniques of image editing. PMID- 15333383 TI - Interactive display of stacks of images in scientific presentations with PowerPoint. AB - OBJECTIVE: We describe a new tool to facilitate the creation and interactive display of stacks of images for scientific and educational presentations using Microsoft PowerPoint. CONCLUSION: This new tool allows interactive scrolling through stacks of images. After a presentation is created, all elements required to display the stacks of images are embedded in the PowerPoint file. These presentations can be run on any computer (Macintosh or IBM-compatible versions) using PowerPoint with no other preinstalled software, making this tool useful for scientific and educational presentations at national and international meetings. PMID- 15333385 TI - Activation of hemostasis after coronary artery bypass grafting with or without cardiopulmonary bypass. AB - Activation of coagulation, fibrinolysis, and the vascular endothelium occurs after heart surgery with cardiopulmonary bypass (CPB), but the effects of eliminating CPB in patients undergoing coronary artery bypass grafting (CABG) are unknown. Therefore, we compared the hemostatic profiles of off-pump and on-pump CABG patients. Two groups of consecutive patients participating in a larger trial (the Octopus Trial) were randomly allocated to undergo CABG with (n = 20) or without (n = 20) CPB. Platelet numbers and plasma concentrations of P-selectin, prothrombin fragment 1.2 (F1.2), soluble fibrin, d-dimers, and von Willebrand factor (as a marker of endothelial cell activation) were measured and corrected for hemodilution. Compared with the on-pump CABG group, F1.2 and d-dimer levels were significantly lower (P = 0.004 and P = 0.03, respectively) in patients having CABG surgery performed off-pump. In the CPB group, F1.2 (median [interquartile range], 450% of baseline [233%-847%]) and d-dimer (538% [318% 1192%]) peaked in the immediate postoperative period and remained increased until Day 4, whereas in the off-pump group, F1.2 and d-dimer levels increased more gradually and peaked on Day 4 (342% [248%-515%] and 555% [387%-882%], respectively). In both groups, von Willebrand factor concentrations were increased until Day 4 (CPB, 308% [228%-405%]; off-pump, 288% [167%-334%]). Despite heparinization, CABG surgery with CPB was associated with excessive thrombin generation and fibrinolytic activity immediately after surgery. The off pump group demonstrated a delayed postoperative response that became equal in magnitude to the CPB in the later (20-96 h) postoperative period. PMID- 15333386 TI - Prevention of cerebral hyperthermia during cardiac surgery by limiting on-bypass rewarming in combination with post-bypass body surface warming: a feasibility study. AB - Cerebral hyperthermia is common during the rewarming phase of cardiopulmonary bypass (CPB) and is implicated in CPB-associated neurocognitive dysfunction. Limiting rewarming may prevent cerebral hyperthermia but risks postoperative hypothermia. In a prospective, controlled study, we tested whether using a surface-warming device could allow limited rewarming from hypothermic CPB while avoiding prolonged postoperative hypothermia (core body temperature <36 degrees C). Thirteen patients undergoing primary elective coronary artery bypass grafting surgery were randomized to either a surface-rewarming group (using the Arctic Sun thermoregulatory system; n = 7) or a control standard rewarming group (n = 6). During rewarming from CPB, the control group was warmed to a nasopharyngeal temperature of 37 degrees C, whereas the surface-warming group was warmed to 35 degrees C, and then slowly rewarmed to 36.8 degrees C over the ensuing 4 h. Cerebral temperature was measured using a jugular bulb thermistor. Nasopharyngeal temperatures were lower in the surface-rewarming group at the end of CPB but not 4 h after surgery. Peak jugular bulb temperatures during the rewarming phase were significantly lower in the surface-rewarming group (36.4 degrees C +/- 1 degrees C) compared with controls (37.7 degrees C +/- 0.5 degrees C; P = 0.024). We conclude that limiting rewarming during CPB, when used in combination with surface warming, can prevent cerebral hyperthermia while minimizing the risk of postoperative hypothermia[corrected]. PMID- 15333387 TI - Does intraoperative evaluation of left ventricular contractile reserve predict myocardial viability? A clinical study using dobutamine stress echocardiography in patients undergoing coronary artery bypass graft surgery. AB - To determine the contractile reserve of the left ventricle during reperfusion as a predictor of myocardial viability in patients undergoing coronary artery bypass graft surgery, we measured the response of left ventricular regional wall motion and thickening by using dobutamine stress echocardiography (DSE) after myocardial revascularization. All patients were monitored with radial and pulmonary arterial catheters, transesophageal echocardiography, standard five-lead clinical electrocardiography, and three-channel Holter electrocardiography. Immediately after separation from cardiopulmonary bypass, dobutamine was administered IV starting at 5 microg. kg(-1). min(-1), with increases in rate every 3 min to 10, 20, 30, and 40 microg. kg(-1). min(-1). Within 1 wk after surgery, resting and redistribution thallium-201 myocardial perfusion imaging (thallium studies) was performed to assess the relationship between the intraoperative contractile response and myocardial viability. One-hundred patients completed DSE up to 10 microg. kg(-1). min(-1), and 85 patients received the larger escalating doses of the DSE. Seventy-two patients had postoperative thallium studies. At the completion of the small-dose dobutamine infusion, 689 (97.7%) of 705 segments had a normal response (improvement), and 16 segments (2.3%) had a positive response (deterioration). During large-dose dobutamine infusion, 577 (95.8%) of 602 segments had a normal response, and 25 segments (4.2%) had a positive response. Myocardial segments that had a positive response during large-dose DSE (48%) were more likely to be considered as nonviable on postoperative thallium studies compared with segments that had a normal response (14.7%) (P < 0.00001). By using thallium studies as the reference standard, the sensitivity of DSE was low (31% and 48% for small- and large-dose DSE, respectively) in predicting nonviable myocardium. However, the specificity was higher (86% and 85% for small- and large dose DSE, respectively). In a separate analysis of patients who developed new regional wall motion abnormalities (RWMA) in the early intraoperative reperfusion period, 15 (75%) of 20 abnormally contracting myocardial segments had normal postoperative thallium studies. Our results demonstrate that a normal response to DSE is highly specific for viable myocardium; however, a positive response to DSE has low sensitivity in predicting nonviable myocardium. The majority of new postbypass regional wall motion abnormalities appear to be related to stunned myocardium. PMID- 15333388 TI - Xenon improves recovery from myocardial stunning in chronically instrumented dogs. AB - In this study we tested the hypothesis that inhalational administration of xenon improves recovery from myocardial stunning. Ten dogs were chronically instrumented for measurement of heart rate; left atrial, aortic, and left ventricular pressure; coronary blood-flow velocity; and myocardial wall thickening fraction. Regional myocardial blood flow was determined with fluorescent microspheres. Catecholamine plasma levels were measured by high performance liquid chromatography. An occluder around the left anterior descending artery (LAD) allowed the induction of a reversible LAD ischemia. Animals underwent 2 experimental conditions in a randomized crossover fashion on separate days: (a) 10 min of LAD occlusion under fentanyl (25 microg. kg(-1). h( 1)) and midazolam (0.6 mg. kg(-1). h(-1)) (control) and (b) a second ischemic episode under the same basal anesthesia with concomitant inhalational administration of 75 +/- 1 vol% xenon (intervention). Anesthesia was induced 35 min before LAD occlusion and was discontinued after 20 min of reperfusion. Dogs receiving xenon showed a significantly better recovery of wall-thickening fraction up to 12 h after ischemia. The increase in plasma epinephrine during emergence from anesthesia and in the early reperfusion period was significantly attenuated in the xenon group. There were no differences between groups concerning global hemodynamics, blood-flow velocity, or regional myocardial blood flow. In conclusion, inhalational administration of 75 vol% xenon improves recovery from myocardial stunning in chronically instrumented dogs under fentanyl/midazolam anesthesia. PMID- 15333389 TI - The effects of high molecular weight hydroxyethyl starch solutions on platelets. AB - Physicochemical characteristics of hydroxyethyl starch (HES) molecules determine their side effects on hemostasis. Our aim in the present experiments was to test the antiplatelet effect of novel high molecular weight HES. Citrated whole blood was hemodiluted in vitro (0% and 20%) with either HES 550 (Hextend), HES 600 (6%Hetastarch-Baxter), HES 200 (Elohast), or the solvent of Hextend in its commercially available solution. The availability of glycoprotein IIb-IIIa was assessed on nonstimulated and on agonist-induced platelets using flow cytometry. Glycoprotein IIb-IIIa availability increased significantly after hemodilution with Hextend and its solvent by 23% and 24%, respectively, but decreased in the presence of 6% Hetastarch-Baxter and Elohast by 18% and 15%, respectively, with no significant difference between the latter two colloids. This study shows that Hextend does not inhibit platelet function as anticipated by its high molecular weight and degree of substitution. The unexpected platelet stimulating effect of Hextend is unique among the currently available HES preparations and may, at least in part, be induced by its solvent containing calcium chloride dihydrate (2.5 mmol/L). The platelet-inhibiting effect of 6%Hetastarch-Baxter was not significantly different from that of medium molecular weight HES 200. PMID- 15333390 TI - Management of an unusual complication during placement of a pulmonary artery catheter. AB - We describe an unusual complication during flotation of a pulmonary artery catheter through a preexisting percutaneous introducer sheath. A malfunctioning pulmonary artery catheter, which was placed through an introducer sheath in the right internal jugular vein, was removed. Attempts at repositioning a second pulmonary artery catheter met with resistance, and we were unable to either advance or withdraw it. Chest radiograph showed a bent introducer sheath going from the right internal jugular vein into the right subclavian vein and a pulmonary artery catheter loop. Under continuous fluoroscopy, the introducer sheath and the pulmonary artery catheter were withdrawn as one unit, which resulted in relaxation of the acute angulation in the introducer sheath and allowed the pulmonary artery catheter to unfold, thus facilitating their complete extraction. We conclude that complications may occur during placement of a pulmonary artery catheter through a well positioned introducer sheath and that fluoroscopy is a valuable tool for safe management of such a complication. PMID- 15333391 TI - Massive pulmonary hemorrhage after pulmonary thromboendarterectomy. AB - Pulmonary thromboendarterectomy, the most common surgical treatment of chronic thromboembolic pulmonary hypertension, is being performed with increasing frequency throughout the world. Massive pulmonary hemorrhage is a potentially fatal complication of this procedure. In this report of three such cases, the diagnosis, clinical course, and possible treatments are discussed. Anesthesiologists involved in the care of patients receiving pulmonary thromboendarterectomy must be aware of the possibility of this complication, as well as the various techniques available for its treatment. PMID- 15333392 TI - The combination of inhaled nitric oxide and pulmonary artery balloon inflation improves oxygenation during whole-lung lavage. AB - We present a case in which the use of inhaled nitric oxide associated with temporary partial unilateral pulmonary artery occlusion further improved oxygenation during whole-lung lavage. This is the first case in the literature describing the simultaneous use of these two modalities. PMID- 15333393 TI - Pheochromocytoma crisis: the use of magnesium sulfate. AB - Pheochromocytoma crisis is a rare life-threatening event that may appear with a variety of clinical symptoms. We present three cases of life-threatening crisis in which magnesium sulfate was particularly beneficial in controlling symptoms and signs when more conventional forms of therapy had failed. Two patients presented with hypertensive encephalopathy, and the third presented with catecholamine-induced cardiomyopathy. All three patients successfully underwent tumor excision with magnesium sulfate used as the sole drug for control of hemodynamic disturbances during surgery. The problems of pheochromocytoma crisis and the potential benefits of magnesium sulfate in this condition are reviewed. PMID- 15333394 TI - The absence of arterial oxygen desaturation during massive oxygen embolism after hydrogen peroxide irrigation. AB - For decades, water-mill murmur, decrease in end-tidal CO(2) (Petco(2)), hypotension, and hypoxemia have been accepted as diagnostic criteria for gas embolism. In this case report, a 19-yr-old male patient developed a sudden reduction in Petco(2) and profound circulatory collapse 15 min after intramedullary irrigation with H(2)O(2). However, arterial oxygen desaturation never developed throughout the entire course of resuscitation from presumed massive oxygen embolism. PMID- 15333395 TI - Subcutaneous tunneling of caudal catheters reduces the rate of bacterial colonization to that of lumbar epidural catheters. AB - Bacterial colonization is regarded as a causative factor for septic complications of caudal catheters in children. To determine whether tunneling caudal catheters reduces the bacterial colonization rate effectively, we evaluated 506 children being treated with tunneled or untunneled caudal or untunneled lumbar epidural catheters. Four-hundred-nine children completed the study. After aseptic removal, the catheters were cultured and sent for microbiological assessment. We found a bacterial colonization rate of 29% in untunneled caudal catheters, 11% in tunneled caudal catheters, and 9% in untunneled lumbar catheters. No severe infectious complications were reported. There was no correlation between catheter retention time and bacterial colonization except for the first 24 h, during which no bacterial colonization was detected. The overall colonization rate remained constant at approximately 13%. We found a positive correlation between bacterial colonization and redness at the catheter entry site. We conclude that tunneled caudal epidural catheters can be used in children for postoperative analgesia without an increased risk of epidural infection. PMID- 15333396 TI - Threshold current for an insulated epidural needle in pediatric patients. AB - We designed this study to determine the threshold current for nerve stimulation of an insulated needle in the epidural space. The intended dermatome was identified using the bony landmarks of the spine. An 18-gauge insulated Tuohy needle was inserted perpendicularly to the skin and advanced until "loss of resistance" was felt. A nerve stimulator was then connected to the insulated needle. Twenty patients were studied using an insulated Tuohy needle and one patient was studied using a noninsulated Tuohy needle. Muscle twitch was elicited with a current of 11.1 +/- 3.1 mA (mean +/- sd) in all patients in which an insulated needle was used. Muscle twitches were within 2 myotomes of the intended level (based on bony landmarks). Muscle twitch was not elicited with a noninsulated needle. After catheter threading, positive stimulation tests were elicited via epidural catheters in all patients (4.9 +/- 2.3 mA). Postoperative radiograph confirmed all catheter placements within 2 myotomes of the muscle twitches. Electrical stimulation may be a useful adjuvant tool to loss of resistance for confirming proper thoracic epidural needle placement. The threshold current criteria for an insulated needle (6-17 mA) would be higher than the original Tsui test criteria described for an epidural catheter (1-10 mA) in the epidural space. PMID- 15333397 TI - The anesthetic management of a patient with Cohen syndrome. AB - Cohen syndrome is a rare genetic disorder caused by autosomal recessive inheritance and is characterized by the following features: mental retardation, infantile hypotonia, micrognathia, narrow and high-arched palate, microcephaly, prominent upper central incisors, poor dentition, short stature, and truncal obesity. Some patients have strabismus, myopia, optic atrophy, and total blindness. A small number of cases present with heart defects or mitral valve prolapse. Only approximately 100 cases have been reported in the world literature. The administration of general anesthesia in patients with Cohen syndrome can be a challenge because most of these patients are mentally retarded and uncooperative and have facial malformations that may make intubation difficult. We present our experience with the anesthetic management of a patient with Cohen syndrome. PMID- 15333398 TI - Reliability and validity of the perioperative opioid-related symptom distress scale. AB - A reduction in opioid use may reduce the incidence and severity of opioid-related side effects. However, no published studies have demonstrated this relationship. In a prospective, placebo-controlled, randomized trial of analgesia for laparoscopic cholecystectomy, we validated an opioid-related symptom distress scale (SDS) questionnaire and clinically meaningful events (CMEs). A total of 193 patients completed the SDS questionnaire every 24 h after discharge for 7 days. This analysis was based on data from Day 1 only. The SDS assessed 12 common opioid-related symptoms, including nausea, vomiting, and difficulty passing urine, by 3 ordinal measures: frequency, severity, and bothersomeness. Patients with responses of "frequently" to "almost constantly," "moderate" to "very severe," or "quite a bit" to "very much bothered" were considered to have a CME. A detailed postoperative recovery survey of patient functional status and experience of adverse effects was used to validate the SDS. Validation measures in the recovery survey were categorized as nonspecific (e.g., level of normal activities) and specific (e.g., number of times vomited in 24 h, minutes of nausea in 24 h, and ability to void normally). SDS scores and CMEs for nausea, vomiting, and difficulty passing urine were strongly associated with three related validation measures from the recovery survey: minutes of nausea within 24 h, number of times vomited within 24 h, and ability to void normally, respectively (P < 0.0001). There was also a strong association between SDS scores and CMEs for nausea, vomiting, and voiding and general recovery validation measures, although the association was significantly weaker than that for symptom specific validation measures. CMEs for nausea, vomiting, and voiding showed a high specificity and lower sensitivity with directly assessed responses. The SDS questionnaire and CMEs are valid tools for assessing postoperative opioid-related symptoms after laparoscopic cholecystectomy. Symptoms defined as CMEs through the SDS may be more sensitive than those identified by direct assessment. PMID- 15333399 TI - Does benzydamine hydrochloride applied preemptively reduce sore throat due to laryngeal mask airway? AB - Sore throat is a common postoperative complaint. We investigated whether preemptive benzydamine hydrochloride (BH) treatment could prevent sore throat due to a laryngeal mask airway (LMA) cuff inflated with air. One-hundred ASA status I II patients who underwent general anesthesia were randomly divided into two groups. In the first group, four puffs of BH were applied to the pharynx 30 min before the operation and 5 min before the induction of anesthesia. Distilled water with a similar bottle was applied with the same protocol in the second group. Anesthetic induction was provided with propofol and fentanyl. The pressure of the LMA cuff inflated with room air was measured after the first adjustment and after 30, 60, and 90 min of inflation in both groups. At the end of operation, the LMA was removed after the recovery of spontaneous breathing. After the operation, patients were asked about sore throat symptoms at the first, second, and fourth hours. There were no significant differences between groups for cuff pressures, cuff volumes, analgesic doses, or operation times. However, sore throat symptoms were significantly less severe for the BH group during both resting and swallowing. In conclusion, preemptive topical BH may decrease the incidence of sore throat due to LMA use. PMID- 15333400 TI - A comparison of intrathecal plain solutions containing ropivacaine 20 or 15 mg versus bupivacaine 10 mg. AB - Ropivacaine, which blocks sensory nerve fibers more readily than motor fibers, is considered to be less potent than bupivacaine. Our hypothesis was that, when used in spinal anesthesia for day surgery, ropivacaine 15 and 20 mg would provide faster motor recovery than bupivacaine 10 mg. This prospective, randomized, double-blinded study included 90 ambulatory lower-extremity surgery patients who received 2 mL of ropivacaine 1%, ropivacaine 0.75%, or bupivacaine 0.5%. Motor block was tested with the Bromage scale, and sensory block was tested with pinprick. Ropivacaine 15 mg provided faster recovery of motor block (150 min) than did bupivacaine 10 mg (210 min; P = 0.005), but the median duration of sensory block at T10 (140 min) did not differ significantly from that with bupivacaine 10 mg (140 min). The median duration of sensory block at T10 was significantly longer with ropivacaine 20 mg (170 min) than with bupivacaine 10 mg (140 min; P = 0.005), but the median recovery from motor block (210 min) did not differ significantly. We conclude that the duration of sensory block of ropivacaine was two thirds and the duration of motor block was half when compared with bupivacaine, with calculations based on the duration-per-milligram of the local anesthetic. PMID- 15333401 TI - Single-dose parenteral pharmacological interventions for the prevention of postoperative shivering: a quantitative systematic review of randomized controlled trials. AB - Shivering is a frequent complication in the postoperative period. The relative efficacy of pharmacological interventions to prevent this phenomenon is not well understood. We performed a systematic search for full reports of randomized comparisons of prophylactic, parenteral, single-dose antishivering interventions with inactive control (placebo or no treatment). Variable doses were converted to fixed doses. Dichotomous data on the absence of shivering were analyzed by using relative benefit (RB) and number needed to treat (NNT) with 95% confidence intervals (CI). Data from 27 trials (1348 adults received an antishivering intervention; 931 were controls) were analyzed. The average incidence of shivering in controls was extremely frequent (52%). Clonidine 65-300 microg (1078 patients), meperidine 12.5-35 mg (250 patients), tramadol 35-220 mg (250 patients), and nefopam 6.5-11 mg (204 patients) were tested in at least 3 trials each. All were more effective than control. For clonidine, meperidine, and nefopam, there was some weak evidence of dose responsiveness. For small-dose clonidine (65-110 microg), the RB compared with control was 1.32 (95% CI, 1.16 1.51); for medium-dose clonidine (140-150 microg), the RB was 1.83 (95% CI, 1.47 2.27); and for large-dose clonidine (220-300 microg), the RB was 1.52 (95% CI, 1.30-1.78). For all clonidine regimens combined, the RB was 1.58 (95% CI, 1.43 1.74), with an NNT of 3.7. For all meperidine regimens combined, the RB was 1.67 (95% CI, 1.37-2.03), with an NNT of 3. For all tramadol regimens combined, the RB was 1.93 (95% CI, 1.56-2.39), with an NNT of 2.2. For all nefopam regimens combined, the RB was 2.62 (95% CI, 2.02-3.40), with an NNT of 1.7. Methylphenidate, midazolam, dolasetron, ondansetron, physostigmine, urapidil, and flumazenil were tested in no more than 3 trials each, with a limited number of patients. PMID- 15333402 TI - The effect of insulin on the resuscitation of bupivacaine-induced severe cardiovascular toxicity in dogs. AB - Resuscitation after bupivacaine-induced cardiovascular collapse is difficult and often resistant to conventional treatment. We tested the hypothesis that insulin treatment would effectively reverse bupivacaine-induced cardiovascular collapse in pentobarbital-anesthetized dogs. Bupivacaine was administered at 0.5 mg. kg( 1). min(-1) until mean arterial blood pressure decreased to 40 mm Hg or less. In the insulin-glucose-potassium (IGK) group (n = 7), an IV bolus of regular insulin (2 U/kg) was given, followed by a glucose infusion (2 mL/kg of 50% dextrose in water) for 30 min and a potassium infusion (1-2 mmol. kg(-1). h(-1)). In the control group (n = 7), glucose infusion was given as in the IGK group. In contrast to the control group, all IGK dogs survived. Mean arterial blood pressure, heart rate, cardiac output, mixed venous oxygen saturation, and end tidal CO(2) recovered toward baseline levels in the IGK group. In conclusion, severe bupivacaine-induced cardiovascular collapse in dogs was effectively reversed with the insulin treatment. PMID- 15333403 TI - Dexamethasone has a central antiemetic mechanism in decerebrated cats. AB - Dexamethasone is an effective antiemetic drug, but its mechanism of action is unclear. We designed this study to investigate the direct antiemetic action of dexamethasone in the medulla of cats. By using an oscillographic vomiting model, decerebrated cats received microinjections of dexamethasone 100 nL (1 microg, n = 7; 0.1 microg, n = 7) into the bilateral nuclei tractus solitarii, which led to a significant prolongation of the latency (1 microg, 6.4 +/- 1.1 min versus 28.2 +/ 4.9 min, P < 0.05; 0.1 microg, 6.7 +/- 1.1 min versus 27.1 +/- 5.0 min, P < 0.05) of the first emetic episode and significantly decreased the frequency of emetic episodes (1 microg, 2.7 +/- 0.8 versus 0.1 +/- 0.4, P < 0.05; 0.1 microg, 2.9 +/- 0.9 versus 0.3 +/- 0.5, P < 0.05) induced by xylazine. Pretreatment with mifepristone, a glucocorticoid receptor antagonist, blocked the antiemetic effect of dexamethasone in the bilateral nuclei tractus solitarii. However, microinjection of dexamethasone into the unilateral nucleus tractus solitarius alone did not alter the latency of the first emetic episode or the frequency of emetic episodes induced by xylazine. Local application of dexamethasone into the area postrema had no effect on the latency of the first emetic episode or the frequency of emetic episodes induced by xylazine. These results suggest that dexamethasone exerts its central antiemetic action through an activation of the glucocorticoid receptors in the bilateral nuclei tractus solitarii in the medulla. PMID- 15333404 TI - Dilution of rocuronium to 0.5 mg/mL with 0.9% NaCl eliminates the pain during intravenous injection in awake patients. AB - In a randomized, double-blinded, controlled study, we evaluated the effect of diluting rocuronium 10 mg/mL to 1 or 0.5 mg/mL with 0.9% NaCl on the pain associated with IV administration of rocuronium with small doses given before succinylcholine or nondepolarizing agent administration. One hundred fifty patients undergoing surgical procedures that required general anesthesia were randomized into three groups. Group 1 received rocuronium 10 mg/mL. Groups 2 and 3 received 1 and 0.5 mg/mL of rocuronium, respectively. Patient demographics, pain scores, osmolality, and the pH of the solutions were recorded. Group 1 had the most intense and frequent pain response. Eighty percent of patients in this group reported pain on injection. In Group 2, the incidence and intensity of pain were significantly less when compared with those of Group 1. In this group, 38% of patients reported pain during injection. In Group 3, none of the patients experienced pain on injection. The pH values and osmolalities of study solutions were not significantly different among groups. In conclusion, in awake patients, dilution of rocuronium 10 mg/mL at small doses given before succinylcholine or nondepolarizing agent administration of 0.06 mg/kg to 0.5 mg/mL with 0.9% NaCl is a simple and cost-effective strategy for preventing pain during IV rocuronium injection. PMID- 15333405 TI - Derivation of preliminary three-dimensional pharmacophores for nonhalogenated volatile anesthetics. AB - We investigated the molecular basis for the immobilizing activity of nonhalogenated volatile anesthetics by using comparative molecular field analysis (CoMFA). In vivo potency data (expressed as minimum alveolar anesthetic concentrations) for 38 structurally diverse drugs were obtained from the literature. The anesthetics were randomly divided into a training-set (n = 28) used to formulate the activity models and a test-set (n = 10) used to independently assess the models' predictive power. The anesthetic structures were aligned to maximize their similarity in molecular shape and electrostatic potential to conformers of the most active drug in the group: hexanol. The individual conformers and alignments with maximum similarity (calculated with combined Carbo indices) were retained and used to derive the CoMFA activity models. The final CoMFA model explained 95.5% of the variance in the observed activities of the training-set anesthetics. The model had good predictive capability for both the training-set drugs (cross-validated r(2) = 0.824) and the randomly excluded test-set anesthetics (r(2) = 0.921). Pharmacophoric maps were derived by identifying the spatial distribution of key areas in which steric and electrostatic interactions are important in determining the immobilizing activity of the anesthetics considered. PMID- 15333406 TI - The soluble guanylyl cyclase inhibitor ODQ, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin 1-one, dose-dependently reduces the threshold for isoflurane anesthesia in rats. AB - Nitric oxide (NO), a cell messenger for activating soluble guanylyl cyclase, is produced by activation of the enzyme NO synthase (NOS) in a wide variety of tissues, including the central nervous system. We have previously demonstrated that inhibition of NOS decreased the minimum alveolar anesthesia concentration (MAC) for isoflurane anesthesia. Moving more distally in the NOS-guanylyl cyclase signaling pathway, we investigated the effects of the specific soluble guanylyl cyclase inhibitor ODQ, 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one, on anesthetic requirements. The effect of ODQ on the MAC of isoflurane anesthesia was investigated in Sprague-Dawley rats while concurrently monitoring the their arterial blood pressure and heart rate. After determining control MAC, ODQ 20-500 mg/kg was administered intraperitoneally 30 min before re-determining MAC in the presence of the soluble guanylyl cyclase inhibitor. In one series, the effect of 250 mg/kg of ODQ on neuronal cyclase guanosine monophosphate production was determined by microdialysis. ODQ produced a statistically significant, dose dependent decrease from isoflurane control MAC (maximal effect 52.4% +/- 2.7%). No ceiling effect was observed over the dose-range studied. This reduction in isoflurane MAC was not accompanied by changes in either heart rate or blood pressure. Inhibition of the NOS-guanylyl cyclase signaling pathway decreased the MAC for isoflurane, which suggests that inhibition of this pathway may play a role in the anesthetic state. The MAC reduction by the soluble guanylyl cyclase inhibitor ODQ was devoid of any significant hemodynamic effects. The current findings, along with the previous observations that structurally distinct NOS inhibitors and the nonspecific soluble guanylyl cyclase inhibitor methylene blue decrease the MAC for volatile anesthetics, support that this is an effect specific to the NOS-guanylyl cyclase signaling pathway. PMID- 15333407 TI - Analysis of gamma-aminobutyric acid-mediated responses in the pulmonary vascular bed of the cat. AB - In this investigation, we sought to identify the role of gamma-aminobutyric acid (GABA)(A) and GABA(B) receptors in the feline pulmonary vascular bed. Using adult mongrel cats and in separate experiments, we investigated the effects of l-N(5) (1-iminoethyl) ornithine hydrochloride (l-NIO) (a nitric oxide synthase inhibitor), glibenclamide (an adenosine triphosphate (ATP)-sensitive K(+) channel blocker), meclofenamate (a nonselective cyclooxygenase inhibitor), bicuculline (a GABA(A) receptor antagonist), and saclofen (a GABA(B) receptor antagonist) on pulmonary arterial responses to pinacidil (an ATP-sensitive K(+) channel activator), bradykinin (a nitric oxide synthase inducer), muscimol (a GABA(A) receptor agonist), and 3-aminopropyl(methyl)phosphinic acid, hydrochloride (SKF 97541; a GABA(B) receptor agonist). Under increased tone conditions in the isolated left lower lobe vascular bed of the cat, muscimol induced a dose dependent vasodepressor response that was not significantly altered after the administration of l-NIO, glibenclamide, meclofenamate, and saclofen. SKF-97541 induced vasodepression was not significantly attenuated after the administration of l-NIO, meclofenamate, and bicuculline. Responses to SKF-97541 were significantly attenuated after the administration of glibenclamide and saclofen. Responses to muscimol were significantly reduced after the administration of bicuculline. The results suggest that muscimol and SKF-97541 have potent vasodepressor activity in the feline pulmonary vascular bed and that these responses are modulated by, respectively, GABA(A) and GABA(B) receptor-sensitive pathways. Further, SKF-97541-induced vasodepression is mediated or modulated by an ATP-sensitive K(+) channel. PMID- 15333408 TI - Local anesthetics adsorbed onto infusion balloon. AB - We compared the adsorption of different local anesthetics onto infusion balloons and studied one of the possible mechanisms for adsorption. After injection of lidocaine, bupivacaine, ropivacaine, and mepivacaine solutions (1 mM each; pH 7.4) into balloons of 100-mL volume, their concentrations in effluents flowing out at 4 mL/h were determined over time by high-performance liquid chromatography. All were adsorbed in a structure-dependent manner, and the concentration decreased by 6%-14% within 5 min. Bupivacaine was most strongly adsorbed, followed by lidocaine, ropivacaine, and mepivacaine. QX-314, a quaternary ammonium derivative of lidocaine, was only weakly adsorbed compared with the parent compound lidocaine. The extent of adsorption of local anesthetics was related to their hydrophobicity (evaluated by reversed-phase chromatography) and was much more at pH 7.4 than at pH 6.0. A hydrophobic interaction with balloon materials appears to be responsible for the adsorption of local anesthetics. When infusion balloons are used for the continuous administration of local anesthetics, attention should be paid to the possibility that their actual concentrations in effluents are smaller than those present when they are initially prepared. PMID- 15333409 TI - Fires from the interaction of anesthetics with desiccated absorbent. AB - Rarely, fire and patient injury have resulted from the degradation of sevoflurane by desiccated carbon dioxide absorbent. Desiccated absorbent also can degrade desflurane and isoflurane, and in the present investigation we sought to determine whether a danger of fire also arose with their use in the presence of desiccated absorbent. Baralyme was desiccated by heating and directing a 10 L/min flow of oxygen through the absorbent. Approximately 1200 g of this desiccated absorbent was used to fill a standard absorber placed in a standard anesthetic circuit to which we directed a 6 L/min flow of oxygen containing 1.5 or 3.0 MAC desflurane, isoflurane, or sevoflurane. A 3-L reservoir bag served as a surrogate lung, and we ventilated this lung with a minute ventilation of 10 L/min. With desflurane or isoflurane, at both 1.5 MAC and 3.0 MAC, temperatures increased in 30 to 70 min to a peak of approximately 100 degrees C and then decreased. With 1.5 MAC sevoflurane (3.0 MAC was not studied), temperatures increased to over 200 degrees C, and in 2 of 5 studies, flames appeared in the anesthetic circuit. In a separate study, we found that concurrent delivery of carbon dioxide and desflurane did not increase peak temperatures. We conclude that the interaction of desflurane or isoflurane with desiccated absorbent is not likely to produce the conflagrations possible with sevoflurane. PMID- 15333410 TI - Ocular microtremor during general anesthesia: results of a multicenter trial using automated signal analysis. AB - Ocular microtremor (OMT) is a fine physiologic tremor of the eye related to neuronal activity in the reticular formation of the brainstem. The frequency of OMT is suppressed by propofol and sevoflurane and predicts the response to command at emergence from anesthesia. Previous studies have relied on post hoc computer analysis of OMT wave forms or on real-time measurements confirmed visually on an oscilloscope. Our overall aim was to evaluate an automated system of OMT signal analysis in a diverse patient population undergoing general anesthesia. In a multicenter trial involving four centers in three countries, we examined the accuracy of OMT to identify the unconscious state and to predict movement in response to airway instrumentation and surgical stimulation. We also tested the effects of neuromuscular blockade and patient position on OMT. We measured OMT continuously by using the closed-eye piezoelectric technique in 214 patients undergoing extracranial surgery with general anesthesia using a variety of anesthetics. OMT decreased at induction in all patients, increased transiently in response to surgical incision or airway instrumentation, and increased at emergence. The frequency of OMT predicted movement in response to laryngeal mask airway insertion and response to command at emergence. Neuromuscular blockade did not affect the frequency of OMT but decreased its amplitude. OMT frequency was unaffected by changes in patient position. We conclude that OMT, measured by an automated signal analysis module, accurately determines the anesthetic state in surgical patients, even during profound neuromuscular blockade and after changes in patient position. PMID- 15333411 TI - The effect of different stages of neuromuscular block on the bispectral index and the bispectral index-XP under remifentanil/propofol anesthesia. AB - Facial electromyographic activity and neuromuscular block could influence bispectral index (BIS) depth of anesthesia monitoring. In this study we examined, in 30 patients undergoing general surgical procedures, the effect of different stages of neuromuscular block on BIS monitoring and compared the conventional A 2000 BIS trade mark (BIS(3.4)) with the new BIS-XP trade mark (BIS(XP)). At deep surgical anesthesia BIS(3.4) of approximately 40, under a propofol 3.61 microg/mL target-controlled infusion and a 0.15-0.3 microg. kg(-1). min(-1) remifentanil infusion, mivacurium 0.15 mg/kg was administered. The onset of neuromuscular block triggered a brief transient odd divergence in response that manifested as a BIS(3.4) increase from 43 +/- 4 to 49 +/- 7 (P = 0.007) and a BIS(XP) decline from 41 +/- 3 to 35 +/- 3 (P = 0.003) at 1 +/- 0.2 min. Then, 2.5 +/- 1 min after mivacurium administration, both monitors returned to baseline values of 43 +/- 5 and 40 +/- 4, respectively. After that, BIS(3.4) and BIS(XP) did not significantly change during complete neuromuscular block or during various levels of neuromuscular recovery. At all phases, BIS(XP) was significantly lower than BIS(3.4). Our study indicated that the BIS(3.4)/BIS(XP) bias and the wide limits of agreement do not allow values given by the two monitors to be used interchangeably. PMID- 15333412 TI - Flow rates and warming efficacy with Hotline and Ranger blood/fluid warmers. AB - The heating capabilities of a water bath blood/fluid warmer, Hotline, have proven superior to those of other devices. The dry heat warmer Ranger has not previously been compared with the Hotline. We evaluated these devices in terms of flow rates and efficacy of warming. We delivered room temperature (21 degrees C) saline and 10 degrees C packed red blood cells (RBCs) by using 90 mm Hg (gravity equivalent) and 300 mm Hg bag pressure and various sizes of IV catheters. The outflow from each device was connected to an inline thermistor, and simultaneous measurements of outflow temperature and flow volume per minute were recorded. Additional data points were obtained with a roller pump that delivered flows of 1-6 L/h through each device. We calculated the effect of these flow rates and outflow temperatures on the mean body temperature (MBT) of a 70-kg patient. The Hotline and Ranger had similar flow rates at 90 and 300 mm Hg pressure infusion when studied with various sizes of IV catheters. Hotline was able to deliver warmer RBCs and saline at slower flow rates (1-4 L/h), but because changes in MBT were almost identical, there was no clinically important advantage, and almost no heat was transferred at these slower flow rates. At more rapid flow rates (>4 L/h), the Ranger warmed RBCs and saline better and produced smaller decreases in MBT than the Hotline. The use of the Hotline for rapid infusions, especially of cold RBCs, is not recommended because of low outflow temperatures and decreases in MBT that were three times larger than those seen with the Ranger. PMID- 15333413 TI - A comparison of the failure times of pulse oximeters during blood pressure cuff induced hypoperfusion in volunteers. AB - Important information may not be obtained if the pulse oximetry signal is lost during inflation of a cuff for blood pressure measurement, particularly in patients with hemodynamic instability. In the present study, we compared the failure times of pulse oximeters during cuff-induced hypoperfusion in volunteers. A pulse oximeter sensor was attached to the index finger, and a blood pressure cuff was attached to the same arm of each volunteer. MasimoSET Radical (Masimo), Nellcor N-395 (N-395), Nellcor N-20PA, and Nellcor D-25 were tested. To evaluate the failure time of each pulse oximeter, time to peak of cuff pressure, time to loss of signal, time to recovery of signal, and failure interval were measured. All measurements were performed three times for each pulse oximeter and were averaged. There were no differences in hemodynamic measurements among the groups. Time to loss of signal was longer in Masimo than the other pulse oximeters. Masimo and N-395 showed significantly shorter times to recovery of signal than those of the other two pulse oximeters. Failure interval was in the order of Masimo << N-395 < Nellcor D-25 = Nellcor N-20PA. Masimo did not lose a signal as rapidly as the other oximeters studied. Masimo was similar in performance to the N-395 at providing useful data sooner than conventional technology after a loss of the signal. These observations suggest that data will be more available with fewer false-positive alarms when using the Masimo oximeter followed by the N-395 when compared with conventional oximeters. PMID- 15333414 TI - Patients' global evaluation of analgesia and safety of injected parecoxib for postoperative pain: a quantitative systematic review. AB - Parecoxib is the only parenterally administered cyclooxygenase-2-selective inhibitor available. We performed a systematic review, including full reports of randomized comparisons of parecoxib compared with any other analgesic intervention for prophylaxis or treatment of postoperative pain. Dichotomous data on patients' global evaluation of their analgesic regimen were extracted by means of the fraction of patients who rated their medication as "good" or "excellent." For safety analysis, data on any reported adverse effects were extracted. Relative risk (RR), number needed to treat (NNT), or number-needed-to-harm were calculated with 95% confidence intervals (CI). Data from 9 trials of 50 initially screened were finally analyzed. One thousand thirteen patients were randomized to receive parecoxib, 218 patients were allocated to an active control, and 507 patients received a placebo. With prophylactic administration, the pooled NNT to obtain the desired outcome ("good"/"excellent" rating) with parecoxib 20 and 40 mg compared with placebo was 4.5 (RR, 1.42; 95% CI, 0.91-2.24) and 4.0 (RR, 1.40; 95% CI, 1.10-1.79), respectively. In the treatment trials, the NNT to obtain the outcome of interest with parecoxib 20 mg was 2.1 (RR, 3.44; 95% CI, 1.49-7.96), 5.3 (RR, 1.43; 95% CI, 1.01-2.02), and -8.3 (RR, 0.85; 95% CI, 0.75-0.97) for the comparisons with placebo, morphine, and ketorolac, respectively. The corresponding NNT for treatment with parecoxib 40 mg was 1.7 (RR, 4.65; 95% CI, 2.04-10.61), 3.7 (RR, 1.62; 95% CI, 1.21-2.16), and 50 (RR, 1.03; 95% CI, 0.89 1.18) for the comparisons with placebo, morphine, and ketorolac, respectively. Overall adverse effects for parecoxib 20 and 40 mg were not different from those with placebo, morphine, or ketorolac. These results suggest a favorable profile for parecoxib compared with inactive or active controls. The optimal dose, timing, and frequency of administration need to be determined. PMID- 15333415 TI - The analgesic efficacy of etoricoxib compared with oxycodone/acetaminophen in an acute postoperative pain model: a randomized, double-blind clinical trial. AB - Our objective in this study was to compare the analgesic effects of etoricoxib and oxycodone/acetaminophen in a postoperative dental pain model. Patients experiencing moderate to severe pain after extraction of two or more third molars were randomized to single doses of etoricoxib 120 mg (n = 100), oxycodone/acetaminophen 10/650 mg (n = 100), or placebo (n = 25). The primary end point was total pain relief over 6 h. Other end-points included patient global assessment of response to therapy; onset, peak, and duration of effect; and rescue opioid analgesic use. Active treatments were statistically significantly superior to placebo for all efficacy measures. Total pain relief over 6 h for etoricoxib was significantly more than for oxycodone/acetaminophen (P < 0.001). Patient global assessment of response to therapy at 6 and 24 h was superior for etoricoxib. Both drugs achieved rapid onset, although the time was faster for oxycodone/acetaminophen by 5 min. The peak effect was similar for both drugs. Compared with oxycodone/acetaminophen patients, etoricoxib patients experienced a longer analgesic duration, had a smaller percentage requiring rescue opioids during 6 and 24 h, and required less rescue analgesia during 6 and 24 h. Oxycodone/acetaminophen treatment resulted in more frequent adverse events (AEs), drug-related AEs, nausea, and vomiting compared with etoricoxib treatment. In conclusion, etoricoxib 120 mg provided superior overall efficacy compared with oxycodone/acetaminophen 10/650 mg and was associated with significantly fewer AEs. PMID- 15333416 TI - The effect of intravenous infusion of adenosine on electrically evoked hyperalgesia in a healthy volunteer model of central sensitization. AB - Human pain models invoking central sensitization, one of the key mechanisms of chronic pain, may be useful for characterizing new analgesics. A new model of electrical hyperalgesia can detect the efficacy of several analgesic mechanisms. Because IV adenosine can alleviate neuropathic pain, we investigated its effect on experimental sensitization. This was a double-blinded, randomized, two-period crossover study in 20 healthy volunteers. Current pulses (0.5 ms; 1 Hz) were applied intracutaneously to achieve pain rating of approximately 5 on a 0-10 numeric rating scale. Pain, areas of pinprick hyperalgesia, and tactile allodynia were assessed during the 2.5-h stimulation period. Adenosine (50 microg. kg(-1). min(-1)) and placebo were infused IV over 60 min. Additional testing was performed 24 h after each treatment. Adenosine reduced the area of pinprick hyperalgesia during the infusion compared with placebo; there was no significant effect on tactile allodynia or pain rating. The effect on hyperalgesia developed over 15 min and was significant (P < or = 0.05) for the rest of the infusion period. There was no difference between treatments at 24 h. Thus, in accordance with reports on neuropathic pain, adenosine reduced central sensitization in the human model of electrical hyperalgesia. However, adenosine did not have the long term effects seen in patients. The model can investigate mechanisms of drugs for the treatment of chronic pain. PMID- 15333417 TI - Inhibition of platelet function by hydroxyethyl starch solutions in chronic pain patients undergoing peridural anesthesia. AB - The use of hydroxyethyl starch (HES) solutions as a fluid replacement before peridural blockade may compromise blood coagulation, thus increasing the risk of neuraxial bleeding. In this prospective, double-blind, placebo-controlled, crossover study, we compared the influence of HES 130 (molecular weight in kilodalton), HES 200, and lactated Ringer's solution on platelet function and hemodynamics in chronic low back pain patients scheduled for peridural blockades. Patients received 3 test infusions of 10 mL/kg each administered IV for 30 min. Collagen/epinephrine and collagen/adenosine diphosphate were used as agonists for assessment of platelet function analyzer-closure times. Arterial blood pressure, heart rate, platelet counts, and hemoglobin levels were documented. Platelet function analyzer-closure times remained stable after lactated Ringer's solution but were significantly prolonged after HES. The platelet-inhibiting effect of HES 200 was more than that of HES 130. Hemodynamic stability was sufficiently maintained by all test infusions. In contrast to previous observations, a relevant antiplatelet effect of both low and medium molecular weight HES solutions was found in this study in chronic pain patients undergoing peridural anesthesia. Because hemostasiological competence is a prerequisite for safe neuraxial blockade, the decision of HES for intravascular fluid administration before blockade should be critically made. PMID- 15333418 TI - Ropivacaine inhibits neurite outgrowth in PC-12 cells. AB - Peripheral nerve injury often leads to neuropathic pain, which might involve sympathetic postganglionic nerve fiber sprouting in the dorsal root ganglion. Recent studies suggest the effectiveness of ropivacaine in blocking neuropathic pain. To determine if ropivacaine affects sympathetic sprouting, we used pheochromocytoma (PC-12) cells, which differentiate into neurons on exposure to nerve growth factor (NGF). PC-12 cells were precultured in the presence of 50 ng/mL of NGF for 4 days. Neurite outgrowth was quantified as neurite extension after 24-, 48-, and 72-h exposure to ropivacaine at doses ranging from 10 to 200 microg/mL. Ropivacaine inhibited the neurite outgrowth in a dose-dependent manner. The inhibitory effect of ropivacaine was completely reversible because the NGF-stimulated neurite outgrowth was recovered to control level after washing out ropivacaine. Ropivacaine, therefore, may exert its therapeutic action on neuropathic pain, at least in part, by suppressing sympathetic sprouting. PMID- 15333419 TI - The incidence of awareness during anesthesia: a multicenter United States study. AB - Awareness with recall after general anesthesia is an infrequent, but well described, phenomenon that may result in posttraumatic stress disorder. There are no recent data on the incidence of this complication in the United States. We, therefore, undertook a prospective study to determine the incidence of awareness with recall during general anesthesia in the United States. This is a prospective, nonrandomized descriptive cohort study that was conducted at seven academic medical centers in the United States. Patients scheduled for surgery under general anesthesia were interviewed in the postoperative recovery room and at least a week after anesthesia and surgery by using a structured interview. Data from 19,575 patients are presented. A total of 25 awareness cases were identified (0.13% incidence). These occurred at a rate of 1-2 cases per 1000 patients at each site. Awareness was associated with increased ASA physical status (odds ratio, 2.41; 95% confidence interval, 1.04-5.60 for ASA status III-V compared with ASA status I-II). Age and sex did not influence the incidence of awareness. There were 46 additional cases (0.24%) of possible awareness and 1183 cases (6.04%) of possible intraoperative dreaming. The incidence of awareness during general anesthesia with recall in the United States is comparable to that described in other countries. Assuming that approximately 20 million anesthetics are administered in the United States annually, we can expect approximately 26,000 cases to occur each year. PMID- 15333420 TI - Do anesthesia information systems increase malpractice exposure? Results of a survey. AB - Anesthesia information systems (AIS) record data from patient monitors and create a detailed electronic anesthesia record. Because the anesthesia record is a primary piece of evidence used in court during a malpractice proceeding, the ability to create an anesthesia record has fostered considerable debate concerning the impact of this method of record keeping on malpractice exposure. Fifty-five departments using AIS were surveyed to document their medicolegal experience with electronic anesthesia records. Twenty-four departments responded to the survey, 18 of which had more than 5 yr experience with the technology. Respondents reported 41 malpractice cases filed since adopting AIS technology. Of the cases filed, 30 were dropped, and 11 went on to settlement or litigation. There were no reported cases in which the automated record hindered the defense process. Eighteen respondents viewed this technology as valuable for risk management, and three more viewed it as essential. The experience reported by these departments indicates that AIS are useful for managing malpractice risk. PMID- 15333421 TI - The effect of lengthening anesthesiology residency on subspecialty education. AB - In this study, we sought to determine the long-term effect of the additional year of anesthesia residency (postgraduate year [PGY]-4) instituted in 1989 by the American Board of Anesthesiology on the number of individuals who pursued 12-mo subspecialty anesthesia training. We tested the hypothesis that extending education by a year would decrease the number of anesthesia subspecialty trainees. Surveys were collected from approved anesthesia residency training programs in the United States from 1989 to 2001. The questionnaires determined the number of individuals pursuing subspecialty training during PGY-4 and PGY-5. The subspecialties included cardiac anesthesia, pediatric anesthesia, pain management, obstetrical anesthesia, neuroanesthesia, outpatient anesthesia, intensive care medicine, and research. The number of anesthesiology residents (PGY-5) pursuing 12-mo subspecialty training increased over this period. The specific subspecialty distribution of fellows changed, with the largest increase in number and percentage occurring in pain management. The largest declines occurred in critical care medicine and research. Our data do not indicate a decrease in the number of anesthesiology subspecialists. Factors other than the duration of training appear responsible for the selection of subspecialty education. PMID- 15333422 TI - The beneficial effects of antioxidant supplementation in enteral feeding in critically ill patients: a prospective, randomized, double-blind, placebo controlled trial. AB - We investigated whether intervention with antioxidant vitamins C and E in enteral feeding influenced oxidative stress and clinical outcome in critically ill patients. Two-hundred-sixteen patients expected to require at least 10 days of enteral feeding completed the study. One-hundred-five patients received enteral feeding supplemented with antioxidants, and 111 control patients received an isocaloric formula. Plasma lipoperoxidation (by thiobarbituric acid reactive substances [TBARS] and prostaglandin F(2alpha) isoprostane levels), low-density lipoprotein (LDL) oxidizability, and LDL tocopherol content were determined at baseline and at the end of the 10-day period. The clinical 28-day outcome was also assessed. Plasma TBARS and isoprostanes were 5.33 +/- 1.26 nM/mL and 312 +/- 68 pg/mL, respectively, before treatment and 2.42 +/- 0.61 nM/mL and 198 +/- 42 pg/mL after intervention (P < 0.01 for both comparisons). Antioxidants improved LDL resistance to oxidative stress by approximately 30% (the lag time before treatment was 87 +/- 23 min and was 118 +/- 20 min after treatment; P < 0.04). There was a significantly reduced 28-day mortality after antioxidant intervention (45.7% in the antioxidant group and 67.5% in the regular-feeding group; P < 0.05). Isoprostanes may provide a sensitive biochemical marker for dose selection in studies involving antioxidants. PMID- 15333423 TI - Bradykinin-induced pulmonary vasoconstriction is time and inducible nitric oxide synthase dependent in a peritonitis sepsis model. AB - In an isolated perfused lung model, bradykinin induced pulmonary vasoconstriction in rats made septic by the injection of lipopolysaccharide (LPS). To mimic the pathophysiology of sepsis in humans more closely, we investigated pulmonary endothelial injury in a peritonitis model (cecal ligation and perforation; CLP). Male Sprague-Dawley rats were randomly divided into nine groups (n = 6-8). LPS and CLP rats were compared after 6 h with and without treatment with a selective inhibitor of inducible nitric oxide synthase (iNOS), L-N(6)-(1-iminoethyl) lysine. Time dependency was investigated in CLP-treated rats at 24 h. The pulmonary circulation was isolated and perfused with a constant flow after the rats' tracheas were intubated and ventilated. Bradykinin (1, 3, and 6 microg) was injected, and changes in perfusion pressure were measured. Lungs were harvested for Western blot analysis to determine the role of iNOS in pulmonary endothelial dysfunction. In contrast to CLP 24 h rats, dose-dependent bradykinin-induced pulmonary vasoconstriction was observed in LPS and CLP 6 h rats. Concomitant administration of L-N(6)-(1-iminoethyl)-lysine significantly attenuated this vasoconstriction in both groups. The iNOS protein was expressed in lung homogenates from LPS 6 h and CLP 6 h but not from CLP 24 h rats. Both sepsis models caused bradykinin-induced pulmonary vasoconstriction, with the CLP groups demonstrating a time dependency of this effect. In conjunction with the time dependent decrease in iNOS protein, the attenuated bradykinin-induced vasoconstriction due to selective iNOS inhibition suggests an important role for iNOS in pulmonary endothelial injury for both sepsis models. PMID- 15333424 TI - Pilot study of the effects of ONO-5046 in patients with acute respiratory distress syndrome. AB - Evidence has linked neutrophil elastase to acute respiratory distress syndrome (ARDS), suggesting that inhibiting the activity of this enzyme could prevent the development and progression of ARDS. However, few clinical trials have examined this notion. We therefore examined the effects of ONO-5046 (sivelestat, a specific inhibitor of neutrophil elastase; sodium N-[2-[4-(2,2 dimethylpropionyloxy) phenylsulfonylaminobenzoyl]amino-acetate tetrahydrate]) in a randomized, double-blinded trial in patients with ARDS. We randomly assigned 24 patients with ARDS to groups that received conventional therapy without or with sivelestat (0.2 mg. kg(-1). h(-1)) for 14 days. The variables of interest associated with clinical outcome were the duration of mechanical ventilation; changes in oxygenation from baseline; changes in cytokine levels from baseline; number of patients alive at 30 days who did not need mechanical ventilation; and mortality rate. The length of intensive care unit stay, number of ventilation days, and mortality rates did not statistically differ between groups. ARDS was more persistent in the control than in the sivelestat group (control, 19.5 +/- 7.4 days; sivelestat, 13.5 +/- 5.9 days; P = 0.039). Neutrophil elastase activity significantly differed between groups at 72 h after treatment. Levels of interleukin-6 were lower in the sivelestat group than in the controls at 24, 48, and 72 h after treatment. ONO-5046 apparently did not affect survival or the duration of mechanical ventilation. PMID- 15333425 TI - Dopexamine reverses the vasopressin-associated impairment in tissue oxygen supply but decreases systemic blood pressure in ovine endotoxemia. AB - Since arginine vasopressin (AVP) may reduce cardiac output and, in proportion, oxygen delivery, we studied the efficacy of dopexamine (DPX) as an adjunct to AVP infusion. After 1 h of continuous AVP infusion (0.04 U/min) in healthy sheep (n = 7), DPX was additionally administered in incremental doses (1, 5, and 10 microg. kg(-1). min(-1); each dose for 30 min). After a 24-h period of recovery, endotoxin was continuously infused in the same sheep to induce and maintain a hypotensive/hyperdynamic circulation. After 16 h of endotoxemia, AVP and DPX were given as described previously. AVP infusion increased systemic vascular resistance index and decreased cardiac index in both healthy and endotoxemic conditions (P < 0.001 each). This was accompanied by an augmented pulmonary vascular resistance index in endotoxemia (159 +/- 13 dynes. cm(-5). m(-2) versus 202 +/- 16 dynes. cm(-5). m(-2)) and a decrease in oxygen delivery index (health: 842 +/- 66 mL. min(-2). m(-2) versus 475 +/- 38 mL. min(-2). m(-2); endotoxemia: 1073 +/- 49 mL. min(-2). m(-2) versus 613 +/- 44 mL. min(-2). m(-2)) and mixed venous oxygen content (health: 63% +/- 2% versus 47% +/- 2%; endotoxemia: 68% +/- 2% versus 51% +/- 3%; P < 0.001 each). Small doses of DPX (1 and 5 microg. kg( 1). min(-1)) improved not only the AVP-associated depressions in cardiac index, oxygen delivery index, and mixed venous oxygen content, but also the pulmonary vasopressive effect in both groups. While large-dose DPX (10 microg. kg(-1). min( 1)) also reduced mean pulmonary arterial pressure in endotoxemia (27 +/- 1 mm Hg versus 23 +/- 1 mm Hg; P < 0.05 versus baseline), mean arterial blood pressure decreased (105 +/- 4 mm Hg versus 80 +/- 3 mm Hg) and heart rate increased (84 +/ 4 bpm versus 136 +/- 9 bpm; P < 0.001 versus AVP alone), thereby limiting its therapeutic use. PMID- 15333426 TI - Closed suctioning system reduces cross-contamination between bronchial system and gastric juices. AB - In this prospective, randomized study, we evaluated whether a closed suctioning (CS) system (TrachCare) influences crossover contamination between bronchial system and gastric juices when compared with an open suctioning system (OS). The secondary aims were an analysis of the frequency of ventilator-associated pneumonia (VAP) and an analysis of alteration in gas exchange. Antibiograms were performed from tracheal secretions and gastric juice aspirates on Days 1 and 3 of intubation in 24 patients in a medical intensive care unit. Five cross contaminations were observed in the OS group on Day 3 versus Day 1; the 5 strains shared common genotypes as determined by random amplification of polymorphic DNA. No cross-contaminations were seen in the CS group (P = 0.037). VAP occurred in 5 patients of the OS group but in none of the CS group patients (P = 0.037). Spao(2) decreased significantly in the OS group compared with presuctioning values--the opposite of the CS group. Whereas presuctioning values were comparable between groups, postsuctioning Spao(2) was significantly higher in the CS group. CS significantly reduced cross-contamination between bronchial system and gastric juices and reduced the incidence of VAP when compared with OS. Hypoxic phases can be reduced by the help of CS. PMID- 15333427 TI - Ketamine for long-term sedation and analgesia of a burn patient. AB - We present a case in which ketamine was used for long-term sedation and analgesia of a burn patient. Under escalating opiate dosages, the patient had developed persistent ileus as well as abdominal distension that caused respiratory compromise, without receiving sufficient analgesia. The opiate-sparing effect of the continuous ketamine infusion was more than 90%. The ileus resolved within 24 h. The quality of sedation also changed favorably. There were no obvious adverse effects of ketamine. PMID- 15333428 TI - Intraischemic nitrous oxide alters neither neurologic nor histologic outcome: a comparison with dizocilpine. AB - N-Methyl-D-aspartate receptor antagonism contributes to the anesthetic action of nitrous oxide (N(2)O). We examined the effects of the N-methyl-D-aspartate antagonists N(2)O and dizocilpine on outcome from filament occlusion of the middle cerebral artery (MCAO). Rats breathed 70% nitrogen/30% oxygen or 70% N(2)O/30% oxygen during MCAO. A third group breathed 70% nitrogen/30% oxygen and was given dizocilpine (0.25 mg/kg IV). After 75 min of MCAO, the rats recovered for 3 or 14 days. Pericranial temperature was maintained at 37.5 degrees C +/- 0.2 degrees C during ischemia and for 20 h postischemia. N(2)O did not alter neurologic scores at 3 days (N(2)O, 21 +/- 6; nitrogen, 22 +/- 8; P = 0.95; 0 = normal; 48 = maximal deficit; mean +/- sd; n = 15) or 14 days (N(2)O, 13 +/- 6; nitrogen, 12 +/- 6; P = 0.93; n = 15-16) postischemia. N(2)O had no effect on infarct size at 3 days (N(2)O, 162 +/- 45 mm(3); nitrogen, 162 +/- 61 mm(3); P > 0.99) or 14 days (N(2)O, 147 +/- 56 mm(3); nitrogen, 151 +/- 62 mm(3); P = 0.99) postischemia. Dizocilpine treatment caused smaller infarcts (3 days: 66 +/- 49 mm(3), P < 0.0001 versus nitrogen; 14 days: 84 +/- 50 mm(3), P < 0.006 versus nitrogen) and reduced the neurologic deficit (3 days: 10 +/- 10, P = 0.002 versus nitrogen; 14 days: 6 +/- 7, P = 0.006 versus nitrogen). N(2)O (70%) had no effect on either behavioral or histologic outcome from transient focal cerebral ischemia when compared with results in rats breathing 70% nitrogen. These results indicate that normobaric N(2)O does not alter the response of rat brain to a focal ischemic insult. PMID- 15333429 TI - Lidocaine impairs vasodilation mediated by adenosine triphosphate-sensitive K+ channels but not by inward rectifier K+ channels in rat cerebral microvessels. AB - Vasodilator effects of adenosine triphosphate (ATP)-sensitive, as well as inward rectifier, K+ channel openers have not been well demonstrated in cerebral microvessels. Although lidocaine impairs vasorelaxation via ATP-sensitive K+ channels in the rat aorta, the effects of this compound on K+ channels in the cerebral circulation have not been shown. We designed the present study to examine whether ATP-sensitive and inward rectifier K+ channels contribute to vasodilator responses in cerebral microvessels and whether the vasodilation mediated by these channels is inhibited by lidocaine. Rat brain slices were monitored using a computer-assisted videomicroscopy. Cerebral parenchymal arterioles (diameter, 5-10 microm) were contracted with prostaglandin F(2alpha), and thereafter potassium chloride (KCl), levcromakalim, or sodium nitroprusside was added to the perfusion chamber. Levcromakalim and KCl produced vasodilation of the cerebral parenchymal arterioles, which was abolished by an ATP-sensitive K+ channel antagonist, glibenclamide, or an inward rectifier K+ channel antagonist, barium chloride, respectively. Lidocaine (10(-5) to 3 x 10(-5) M) inhibited the dilation produced by levcromakalim but not by KCl or sodium nitroprusside. In parenchymal arterioles of the cerebral cortex, lidocaine seems to reduce vasodilation mediated by ATP-sensitive K+ channels but not by inward rectifier K+ channels. PMID- 15333430 TI - Pulmonary edema induced by calcium-channel blockade for tocolysis. AB - Nicardipine is used in the treatment of premature labor. There are no previous reports in the anesthesia literature of serious side effects associated with this drug. We report a case of pulmonary edema induced by nicardipine therapy for tocolysis in a pregnant 27-yr-old patient admitted to our hospital for preterm labor with intact membranes at 27 wk of gestation. PMID- 15333431 TI - Bupivacaine in microcapsules prolongs analgesia after subcutaneous infiltration in humans: a dose-finding study. AB - In this study, we examined the onset and duration of local analgesic effects of bupivacaine incorporated into biodegradable microcapsules (extended-duration local anesthetic; EDLA) administered as subcutaneous infiltrations in different doses in humans. In 18 volunteers, the skin on the medial calf was infiltrated with 10 mL of EDLA, and the opposite calf was infiltrated with 10 mL of aqueous bupivacaine (5.0 mg/mL) in a double-blinded, randomized manner. Three different concentrations of EDLA were tested (6.25, 12.5, and 25 mg/mL), with 6 subjects in each group. Pain responses to mechanical and heat stimuli and sensory thresholds (touch, warm, and cold detection thresholds) were examined by von Frey hairs and contact thermodes. Assessments were made before and 2, 4, 6, 8, 24, 48, 72, 96, and 168 h after the injections. Safety evaluations were performed daily for the first week and at 2 wk, 6 wk, and 6 mo after the injections. The time to maximum effects was significantly shorter for aqueous bupivacaine (2-6 h) than for EDLA (4-24 h), but there were no significant differences between the maximum effects of EDLA and aqueous bupivacaine. From 24 to 96 h after the injections, EDLA was significantly more efficient than aqueous bupivacaine for all variables, and significant effects of EDLA were demonstrated for at least 96 h for all variables. In general, a dose-response gradient was seen in the EDLA group for 5 of 7 variables when the curves expressing effect over time for the different concentrations were evaluated. No serious side effects were observed for up to 6 mo after administration. In conclusion, bupivacaine incorporated in microcapsules provided analgesia for 96 h after subcutaneous infiltration. PMID- 15333432 TI - Levobupivacaine 0.2% or 0.125% for continuous sciatic nerve block: a prospective, randomized, double-blind comparison with 0.2% ropivacaine. AB - In 60 patients receiving elective hallux valgus repair, we compared the efficacy of continuous popliteal sciatic nerve block produced with 0.2% ropivacaine (n = 20), 0.2% levobupivacaine (n = 20), or 0.125% levobupivacaine (n = 20) infused with a patient-controlled system starting 3 h after a 30-mL bolus of the 0.5% concentration of the study drug and for 48 h (baseline infusion rate, 6 mL/h; incremental dose, 2 mL; lockout time, 15 min; maximum incremental doses per hour, 3). No differences were reported in the intraoperative efficacy of the nerve block. The degree of pain was similar in the three groups throughout the study period, both at rest and during motion. Total consumption of local anesthetic solution during the first 24 h was 148 mL (range, 144-228 mL) with 0.2% ropivacaine, 150 mL (range, 144-200 mL) with 0.2% levobupivacaine, and 148 mL (range, 144-164 mL) with 0.125% levobupivacaine (P = 0.59). The volume of local anesthetic consumed during the second postoperative day was 150 mL (range, 144 164 mL) with 0.2% ropivacaine, 154 mL (range, 144-176 mL) with 0.2% levobupivacaine, and 151 mL (range, 144-216 mL) with 0.125% levobupivacaine (P = 0.14). A smaller proportion of patients receiving 0.2% levobupivacaine showed complete recovery of foot motor function as compared with 0.2% ropivacaine and 0.125% levobupivacaine, both at 24 h (35% vs 85% and 95%; P = 0.0005) and at 48 h (60% vs 100% and 100%; P = 0.001). We conclude that sciatic infusion with both 0.125% and 0.2% levobupivacaine provides adequate postoperative analgesia after hallux valgus repair, clinically similar to that provided by 0.2% ropivacaine; however, the 0.125% concentration is preferred if early mobilization of the operated foot is required. PMID- 15333433 TI - The effects of cervical and lumbar epidural anesthesia on heart rate variability and spontaneous sequence baroreflex sensitivity. AB - A high level of neuroaxial block may produce profound bradycardia and hypotension, possibly as a result of an imbalance between sympathetic and parasympathetic control of heart rate. We designed this study to test the hypothesis that cervical epidural anesthesia would increase the high-frequency (HF) component of heart rate variability (HRV) as a result of cardiac sympathectomy, whereas lumbar epidural anesthesia would cause sympathetic predominance. HRV and spontaneous baroreflex (SBR) sensitivity were assessed before and after cervical and lumbar epidural anesthesia by using plain 1.5% lidocaine (median upper/lower sensory block: C3/T8 for cervical and T11/L5 for lumbar) in healthy patients (n = 10 each). Electrocardiogram and noninvasive beat to-beat arterial blood pressure were monitored. HRV was analyzed by using fast Fourier transformation. Least-square regression analysis relating R-R interval and systolic blood pressure during spontaneous fluctuation was performed to obtain SBR sensitivities. Cervical epidural group patients were significantly older (P < 0.01) and taller (P < 0.01). Cervical epidural anesthesia attenuated HF (0.15-0.4 Hz) and low-frequency (0.04-0.15 Hz) power of HRV with concomitant reductions in up- and down-sequence SBR sensitivities, suggesting decreased vagal modulation of heart rate. Lumbar epidural anesthesia resulted in a significant increase in the low-frequency/HF ratio of HRV and unchanged SBR indices, suggesting sympathetic predominance. HF power correlated well with SBR sensitivities under most of our study conditions. Respiratory rates and Paco(2) were unchanged by either epidural technique. Our results indicate that cervical, but not lumbar, epidural anesthesia depresses phasic and tonic dynamic modulation of the cardiac cycle by the vagal nerve in conscious humans. PMID- 15333434 TI - Local anesthetics inhibit thromboxane A2 signaling in Xenopus oocytes and human k562 cells. AB - Thromboxane A(2) (TXA(2)) has been proposed as a mediator of perioperative myocardial ischemia, vasoconstriction, and thrombosis. As these adverse events are minimized with epidural anesthesia, rather than general anesthesia, we hypothesized that local anesthetics would inhibit TXA(2)-receptor signaling. We used fluorometric determination of intracellular [Ca(2+)] in human K562 cells and 2-electrode voltage clamp measurements in Xenopus laevis oocytes expressing TXA(2) receptors. After 10-min incubation, lidocaine (IC(50): 1.02 +/- 0.2 x 10( 3) M), ropivacaine (IC(50): ropivacaine 6.3 +/- 0.9 x 10(-5) M), or bupivacaine (IC(50): 1.42 +/- 0.08 x 10(-7) M) inhibited TXA(2)-induced [Ca(2+)](i) in K562 cells. These data were confirmed in Xenopus oocytes recombinantly expressing TXA(2) receptors, with IC(50)s of bupivacaine 1.2 +/- 0.2 x 10(-5) M, R(+) ropivacaine 4.9 +/- 1.7 x 10(-4) M, S(-) ropivacaine 5.3 +/- 0.9 x 10(-5) M, and lidocaine 6.4 +/- 2.8 x 10(-4) M. Intracellular pathways activated by IP(3) and GTPgammaS were not significantly affected by the local anesthetics tested. QX314, a positively charged lidocaine analog, inhibited only if injected intracellularly (IC(50): 5.3 +/- 1.7 x 10(-4) M), indicating one local anesthetic target is most likely inside the cell. Benzocaine (largely uncharged) inhibited with an IC(50) of 8.7 +/- 1.8 x 10(-4) M. This suggests that some of the beneficial effects of regional anesthesia techniques might be due to direct interaction of local anesthetics with the functioning of membrane proteins. PMID- 15333435 TI - Effective nonanatomical endoscopy training produces clinical airway endoscopy proficiency. AB - We studied the effectiveness of two nonanatomical endoscopic dexterity training models: "Choose the Hole" and Dexter. Effectiveness was assessed in terms of time spent training, subjective rating, performance on an anatomical manikin, and clinical performance on fellow participants who acted as awake subjects. Forty three anesthesia specialists, trainees, and technicians volunteered. Performances were videotaped, timed, and scored with a Global Rating Scale (GRS) from 1 (very poor) to 5 (clearly superior). The Dexter group spent more time training than the Choose the Hole group (median time [range], 152 min [70-510 min] versus 75 min [17-281 min]; P < 0.01). Subjective ratings were better in the Dexter group. In clinical bronchoscopy, the Dexter group was faster (30.7 s [17.1-43.5 s] versus 36.6 s [22.8-105.1 s]; P = 0.02) and had higher GRS scores (mean [sd]: 3.0 [0.4] versus 2.6 [0.6]; P = 0.04), indicating superior performance. Clinical and manikin performance (GRS scores) were significantly correlated (rho = 0.62; P = 0.0001). Benchmark levels of clinical bronchoscopic performance can be anticipated from bench model performance without a clinical learning curve. Dexter is a more effective model for learning endoscopic dexterity than the Choose the Hole model. Airway topicalization with lidocaine in a dose range consistent with published series (490-980 mg or 7.14-14.77 mg/kg) resulted in a frequent incidence of side effects. No major adverse events occurred. PMID- 15333436 TI - Sequential one-lung isolation using a double arndt bronchial blocker technique. AB - One-lung ventilation and isolation can be performed using a double-lumen endotracheal tube or one of several alternative airway devices. We report a case in which sequential lung isolation was performed by placing two Arndt bronchial blockers through a single-lumen endotracheal tube. Traditional double-lumen tubes can be difficult to place and have potential complications that may be avoided by placing two Arndt blockers. PMID- 15333437 TI - Antiplatelet and anticoagulant prophylaxis and postoperative blood loss in cardiac surgery. PMID- 15333438 TI - Antiplatelet agents and bleeding after cardiac surgery. PMID- 15333439 TI - AEP "Click Detection" failure: may be, may be not! PMID- 15333440 TI - A simple technique to properly position a Swan Ganz catheter in cardiac surgical patients with situs inversus. PMID- 15333441 TI - A novel approach to infraclavicular brachial plexus block: the ultrasound experience. PMID- 15333442 TI - Warm steaming increases the analgesic effect of topical lidocaine tape on venous cannulation. PMID- 15333443 TI - Partial airway obstruction by a pediatric laryngeal mask airway. PMID- 15333444 TI - Tracheal intubation in the lateral position. PMID- 15333445 TI - Reducing venipuncture pain by cough trick. PMID- 15333446 TI - Don't use thiopental to decrease propofol injection pain. PMID- 15333447 TI - Postoperative analgesia following total knee arthroplasty. PMID- 15333448 TI - Interpretation of PO2 via FIO2. PMID- 15333449 TI - Standard clinical tests for predicting difficult intubation are not useful among morbidly obese patients. PMID- 15333450 TI - Let us not displace small-dose dopamine from the modern ICU so soon. PMID- 15333451 TI - Implications of the incidence of influenza-like illness in nursing homes for influenza chemoprophylaxis: descriptive study. PMID- 15333452 TI - Effect of lactobacillus in preventing post-antibiotic vulvovaginal candidiasis: a randomised controlled trial. AB - OBJECTIVE: To test whether oral or vaginal lactobacillus can prevent vulvovaginitis after antibiotic treatment. DESIGN: Randomised, placebo controlled, double blind, factorial 2x2 trial. SETTING: Fifty general practices and 16 pharmacies in Melbourne, Australia. PARTICIPANTS: Non-pregnant women aged 18-50 years who required a short course of oral antibiotics for a non gynaecological infection: 278 were enrolled in the study, and results were available for 235. INTERVENTIONS: Lactobacillus preparations taken orally or vaginally, or both, from enrollment until four days after completion of their antibiotic course. MAIN OUTCOME MEASURES: Participants' reports of symptoms of post-antibiotic vulvovaginitis, with microbiological evidence of candidiasis provided by a self obtained vaginal swab. RESULTS: Overall, 55/235 (23% (95% confidence interval 18% to 29%)) women developed post-antibiotic vulvovaginitis. Compared with placebo, the odds ratio for developing post-antibiotic vulvovaginitis with oral lactobacillus was 1.06 (95% confidence interval 0.58 to 1.94) and with vaginal lactobacillus 1.38 (0.75 to 2.54). Compliance with antibiotics and interventions was high. The trial was terminated after the second interim analysis because of lack of effect of the interventions. Given the data at this time, the chances of detecting a significant reduction in vulvovaginitis with oral or vaginal lactobacillus treatment were less than 0.032 and 0.0006 respectively if the trial proceeded to full enrollment. CONCLUSIONS: The use of oral or vaginal forms of lactobacillus to prevent post-antibiotic vulvovaginitis is not supported by these results. Further research on this subject is unlikely to be fruitful, unless new understandings about the pathogenesis of post antibiotic vulvovaginitis indicate a possible role for lactobacillus. PMID- 15333453 TI - DWE: discriminating word enumerator. AB - MOTIVATION: Tissue-specific transcription factor binding sites give insight into tissue-specific transcription regulation. RESULTS: We describe a word-counting based tool for de novo tissue-specific transcription factor binding site discovery using expression information in addition to sequence information. We incorporate tissue-specific gene expression through gene classification to positive expression and repressed expression. We present a direct statistical approach to find overrepresented transcription factor binding sites in a foreground promoter sequence set against a background promoter sequence set. Our approach naturally extends to synergistic transcription factor binding site search. We find putative transcription factor binding sites that are overrepresented in the proximal promoters of liver-specific genes relative to proximal promoters of liver-independent genes. Our results indicate that binding sites for hepatocyte nuclear factors (especially HNF-1 and HNF-4) and CCAAT/enhancer-binding protein (C/EBPbeta) are the most overrepresented in proximal promoters of liver-specific genes. Our results suggest that HNF-4 has strong synergistic relationships with HNF-1, HNF-4 and HNF-3beta and with C/EBPbeta. AVAILABILITY: Programs are available for use over the Web at http://rulai.cshl.edu/tools/dwe. PMID- 15333454 TI - HapBlock: haplotype block partitioning and tag SNP selection software using a set of dynamic programming algorithms. AB - Recent studies have revealed that linkage disequilibrium (LD) patterns vary across the human genome with some regions of high LD interspersed with regions of low LD. Such LD patterns make it possible to select a set of single nucleotide polymorphism (SNPs; tag SNPs) for genome-wide association studies. We have developed a suite of computer programs to analyze the block-like LD patterns and to select the corresponding tag SNPs. Compared to other programs for haplotype block partitioning and tag SNP selection, our program has several notable features. First, the dynamic programming algorithms implemented are guaranteed to find the block partition with minimum number of tag SNPs for the given criteria of blocks and tag SNPs. Second, both haplotype data and genotype data from unrelated individuals and/or from general pedigrees can be analyzed. Third, several existing measures/criteria for haplotype block partitioning and tag SNP selection have been implemented in the program. Finally, the programs provide flexibility to include specific SNPs (e.g. non-synonymous SNPs) as tag SNPs. AVAILABILITY: The HapBlock program and its supplemental documents can be downloaded from the website http://www.cmb.usc.edu/~msms/HapBlock. PMID- 15333455 TI - Comment on 'Discrepancies in dbSNP confirmations rates and allele frequency distributions from varying genotyping error rates and patterns'. PMID- 15333456 TI - SABmark--a benchmark for sequence alignment that covers the entire known fold space. AB - The Sequence Alignment Benchmark (SABmark) provides sets of multiple alignment problems derived from the SCOP classification. These sets, Twilight Zone and Superfamilies, both cover the entire known fold space using sequences with very low to low, and low to intermediate similarity, respectively. In addition, each set has an alternate version in which unalignable but apparently similar sequences are added to each problem. PMID- 15333457 TI - OntologyTraverser: an R package for GO analysis. AB - Gene Ontology (GO) annotations have become a major tool for analysis of genome scale experiments. We have created OntologyTraverser--an R package for GO analysis of gene lists. Our system is a major advance over previous work because (1) the system can be installed as an R package, (2) the system uses Java to instantiate the GO structure and the SJava system to integrate R and Java and (3) the system is also deployed as a publicly available web tool. AVAILABILITY: Our software is academically available through http://franklin.imgen.bcm.tmc.edu/OntologyTraverser/. Both the R package and the web tool are accessible. CONTACT: cashaw@bcm.tmc.edu PMID- 15333458 TI - Gene name ambiguity of eukaryotic nomenclatures. AB - MOTIVATION: With more and more scientific literature published online, the effective management and reuse of this knowledge has become problematic. Natural language processing (NLP) may be a potential solution by extracting, structuring and organizing biomedical information in online literature in a timely manner. One essential task is to recognize and identify genomic entities in text. 'Recognition' can be accomplished using pattern matching and machine learning. But for 'identification' these techniques are not adequate. In order to identify genomic entities, NLP needs a comprehensive resource that specifies and classifies genomic entities as they occur in text and that associates them with normalized terms and also unique identifiers so that the extracted entities are well defined. Online organism databases are an excellent resource to create such a lexical resource. However, gene name ambiguity is a serious problem because it affects the appropriate identification of gene entities. In this paper, we explore the extent of the problem and suggest ways to address it. RESULTS: We obtained gene information from 21 organisms and quantified naming ambiguities within species, across species, with English words and with medical terms. When the case (of letters) was retained, official symbols displayed negligible intra species ambiguity (0.02%) and modest ambiguities with general English words (0.57%) and medical terms (1.01%). In contrast, the across-species ambiguity was high (14.20%). The inclusion of gene synonyms increased intra-species ambiguity substantially and full names contributed greatly to gene-medical-term ambiguity. A comprehensive lexical resource that covers gene information for the 21 organisms was then created and used to identify gene names by using a straightforward string matching program to process 45,000 abstracts associated with the mouse model organism while ignoring case and gene names that were also English words. We found that 85.1% of correctly retrieved mouse genes were ambiguous with other gene names. When gene names that were also English words were included, 233% additional 'gene' instances were retrieved, most of which were false positives. We also found that authors prefer to use synonyms (74.7%) to official symbols (17.7%) or full names (7.6%) in their publications. CONTACT: lifeng.chen@dbmi.columbia.edu PMID- 15333459 TI - A new algorithm for detecting low-complexity regions in protein sequences. AB - MOTIVATION: Pair-wise alignment of protein sequences and local similarity searches produce many false positives because of compositionally biased regions, also called low-complexity regions (LCRs), of amino acid residues. Masking and filtering such regions significantly improves the reliability of homology searches and, consequently, functional predictions. Most of the available algorithms are based on a statistical approach. We wished to investigate the structural properties of LCRs in biological sequences and develop an algorithm for filtering them. RESULTS: We present an algorithm for detecting and masking LCRs in protein sequences to improve the quality of database searches. We developed the algorithm based on the complexity analysis of subsequences delimited by a pair of identical, repeating subsequences. Given a protein sequence, the algorithm first computes the suffix tree of the sequence. It then collects repeating subsequences from the tree. Finally, the algorithm iteratively tests whether each subsequence delimited by a pair of repeating subsequences meets a given criteria. Test results with 1000 proteins from 20 families in Pfam show that the repeating subsequences are a good indicator for the low-complexity regions, and the algorithm based on such structural information strongly compete with others. AVAILABILITY: http://bioinfo.knu.ac.kr/research/CARD/ CONTACT: swshin@bioinfo.knu.ac.kr PMID- 15333460 TI - Filtering of ineffective siRNAs and improved siRNA design tool. AB - MOTIVATION: Short interfering RNAs (siRNAs) can be used to suppress gene expression and possess many potential applications in therapy, but how to design an effective siRNA is still not clear. Based on the MPI (Max-Planck-Institute) basic principles, a number of siRNA design tools have been developed recently. The set of candidates reported by these tools is usually large and often contains ineffective siRNAs. In view of this, we initiate the study of filtering ineffective siRNAs. RESULTS: The contribution of this paper is 2-fold. First, we propose a fair scheme to compare existing design tools based on real data in the literature. Second, we attempt to improve the MPI principles and existing tools by an algorithm that can filter ineffective siRNAs. The algorithm is based on some new observations on the secondary structure, which we have verified by AI techniques (decision trees and support vector machines). We have tested our algorithm together with the MPI principles and the existing tools. The results show that our filtering algorithm is effective. AVAILABILITY: The siRNA design software tool can be found in the website http://www.cs.hku.hk/~sirna/ CONTACT: smyiu@cs.hku.hk PMID- 15333461 TI - Missing value estimation for DNA microarray gene expression data: local least squares imputation. AB - MOTIVATION: Gene expression data often contain missing expression values. Effective missing value estimation methods are needed since many algorithms for gene expression data analysis require a complete matrix of gene array values. In this paper, imputation methods based on the least squares formulation are proposed to estimate missing values in the gene expression data, which exploit local similarity structures in the data as well as least squares optimization process. RESULTS: The proposed local least squares imputation method (LLSimpute) represents a target gene that has missing values as a linear combination of similar genes. The similar genes are chosen by k-nearest neighbors or k coherent genes that have large absolute values of Pearson correlation coefficients. Non parametric missing values estimation method of LLSimpute are designed by introducing an automatic k-value estimator. In our experiments, the proposed LLSimpute method shows competitive results when compared with other imputation methods for missing value estimation on various datasets and percentages of missing values in the data. AVAILABILITY: The software is available at http://www.cs.umn.edu/~hskim/tools.html CONTACT: hpark@cs.umn.edu PMID- 15333462 TI - Recombination Analysis Tool (RAT): a program for the high-throughput detection of recombination. AB - MOTIVATION: Recombination can be a prevailing drive in shaping genome evolution. RAT (Recombination Analysis Tool) is a Java-based tool for investigating recombination events in any number of aligned sequences (protein or DNA) of any length (short viral sequences to full genomes). It is an uncomplicated and intuitive application and allows the user to view only the regions of sequence alignments they are interested in. RESULTS: RAT was applied to viral sequences. Its utility was demonstrated through the detection of a known recombinant of HIV and a detailed analysis of Noroviruses, the most common cause of viral gastroenteritis in humans. AVAILABILITY: RAT, along with a user's guide, is freely available from http://jic-bioinfo.bbsrc.ac.uk/bioinformatics research/staff/graham_etherington/RAT.htm. PMID- 15333463 TI - Dual-genome primer design for construction of DNA microarrays. AB - MOTIVATION: Microarray experiments using probes covering a whole transcriptome are expensive to initiate, and a major part of the costs derives from synthesizing gene-specific PCR primers or hybridization probes. The high costs may force researchers to limit their studies to a single organism, although comparing gene expression in different species would yield valuable information. RESULTS: We have developed a method, implemented in the software DualPrime, that reduces the number of primers required to amplify the genes of two different genomes. The software identifies regions of high sequence similarity, and from these regions selects PCR primers shared between the genomes, such that either one or, preferentially, both primers in a given PCR can be used for amplification from both genomes. To assure high microarray probe specificity, the software selects primer pairs that generate products of low sequence similarity to other genes within the same genome. We used the software to design PCR primers for 2182 and 1960 genes from the hyperthermophilic archaea Sulfolobus solfataricus and Sulfolobus acidocaldarius, respectively. Primer pairs were shared among 705 pairs of genes, and single primers were shared among 1184 pairs of genes, resulting in a saving of 31% compared to using only unique primers. We also present an alternative primer design method, in which each gene shares primers with two different genes of the other genome, enabling further savings. 3. AVAILABILITY: The software is freely available at http://www.biotech.kth.se/molbio/microarray/. PMID- 15333464 TI - Ozone effects on the ultrastructure of peatland plants: Sphagnum mosses, Vaccinium oxycoccus, Andromeda polifolia and Eriophorum vaginatum. AB - BACKGROUND AND AIMS: Ozone effects on peatland vegetation are poorly understood. Since stress responses are often first visible in cell ultrastructure, electron microscopy was used to assess the sensitivity of common peatland plants to elevated ozone concentrations. METHODS: Three moss species (Sphagnum angustifolium, S. magellanicum and S. papillosum), a graminoid (Eriophorum vaginatum) and two dwarf shrubs (Vaccinium oxycoccus and Andromeda polifolia), all growing within an intact canopy on peat monoliths, were exposed to a concentration of 0, 50, 100 or 150 ppb ozone in two separate growth chamber experiments simulating either summer or autumn conditions in central Finland. After a 4- or 5-week-long exposure, samples were photographed in a transmission electron microscope and analysed quantitatively using image processing software. KEY RESULTS: In the chlorophyllose cells of the Sphagnum moss leaves from the capitulum, ozone exposure led to a decrease in chloroplast area and in granum stack thickness and various changes in plastoglobuli and cell wall thickness, depending on the species and the experiment. In E. vaginatum, ozone exposure significantly reduced chloroplast cross-sectional areas and the amount of starch, whereas there were no clear changes in the plastoglobuli. In the dwarf shrubs, ozone induced thickening of the cell wall and an increase in the size of plastoglobuli under summer conditions. In contrast, under autumn conditions the cell wall thickness remained unchanged but ozone exposure led to a transient increase in the chloroplast and starch areas, and in the number and size of plastoglobuli. CONCLUSIONS: Ozone responses in the Sphagnum mosses were comparable to typical ozone stress symptoms of higher plants, and indicated sensitivity especially in S. angustifolium. The responses in the dwarf shrubs suggest stimulation of photosynthesis by low ozone concentrations and ozone sensitivity only under cool autumn conditions. PMID- 15333465 TI - Specific combinations of DNA repair gene variants and increased risk for non small cell lung cancer. AB - Several polymorphisms in DNA repair genes have been reported to be associated with lung cancer risk including XPA (-4G/A), XPD (Lys751Gln and Asp312Asn), XRCC1 (Arg399Gln), APE1 (Asp148Glu) and XRCC3 (Thr241Met). As there is little information on the combined effects of these variants, polymorphisms were analyzed in a case-control study including 463 lung cancer cases [among them 204 adenocarcinoma and 212 squamous cell carcinoma (SCC)] and 460 tumor-free hospital controls. Odds ratios (OR) adjusted for age, gender, smoking and occupational exposure were calculated for the variants alone and combinations thereof. For homozygous individuals carrying the Glu variant of APE1, a protective effect was found (OR = 0.77, CI = 0.51-1.16). Individuals homozygous for the variants XPA ( 4A) (OR = 1.53, CI = 0.94-2.5), XPD 751Gln (OR = 1.39, CI = 0.90-2.14) or XRCC3 241Met (OR = 1.29, CI = 0.85-1.98) showed a slightly higher risk for lung cancer overall. In the subgroup of adenocarcinoma cases, adjusted ORs were increased for individuals homozygous for XPA (-4A) (OR = 1.62, CI = 0.91-2.88) and XRCC3 241Met (OR = 1.65; CI = 0.99-2.75). When analyzing the combined effects of variant alleles, 54 patients and controls were identified that were homozygous for two or three of the potential risk alleles [i.e. the variants in nucleotide excision repair, XPA (-4A) and XPD 751Gln, and in homologous recombination, XRCC3-241Met]. ORs were significantly increased when all patients (OR = 2.37; CI = 1.26-4.48), patients with SCC (OR = 2.83; CI = 1.17-6.85) and with adenocarcinoma (OR = 3.05; CI = 1.49-6.23) were analyzed. Combinations of polymorphisms in genes involved in the same repair pathway (XPA + XPD or XRCC1 + APE1) affected lung cancer risk only in patients with SCC. These results indicate that lung cancer risk is only moderately increased by single DNA repair gene variants investigated but it is considerably enhanced by specific combinations of variant alleles. Analyses of additional DNA repair gene interactions in larger population-based studies are warranted for identification of high-risk subjects. PMID- 15333466 TI - Association of matrilysin-2 (MMP-26) expression with tumor progression and activation of MMP-9 in esophageal squamous cell carcinoma. AB - Expression of matrilysin-2, matrix metalloproteinase (MMP)-26, has been implicated in the progression of several types of human cancer. Matrilysin-2 has been reported to be a physiological and pathological activator of pro-MMP-9. The aim of this study was to examine matrilysin-2 expression and determine whether it is correlated with progression of human esophageal squamous cell carcinoma (ESCC). Semi-quantitative reverse transcriptase-polymerase chain reaction, immunohistochemical analysis, zymography and an in vitro invasion assay were performed. Matrilysin-2 mRNA expression was undetectable or only faintly detected in non-tumor tissues, but its overexpression was detected in 24 of the 50 ESCC tissues. Matrilysin-2 overexpression was significantly correlated with depth of invasion, lymph node metastasis and an advance in pathological tumor node metastasis (pTNM) stage. Sections with immunostaining signals in >10% of carcinoma cells at the invasive front, which were observed in 46 of 100 cases, were judged to be positive for matrilysin-2 expression. Matrilysin-2 expression was significantly correlated with depth of invasion, lymph node and distant metastasis, advance in pTNM stage and recurrence. Expression of matrilysin-2 was significantly correlated with nuclear beta-catenin expression and MMP-9 expression. Patients with matrilysin-2-positive cancer had significantly shorter overall and disease-free survival periods than did those with matrilysin-2 negative cancer. Matrilysin-2 expression retained its significant predictive value for overall and disease-free survival in multivariate analysis. Moreover, patients with concomitant expression of matrilysin-2 and MMP-9 had the worst prognosis. Zymography revealed that matrilysin-2 expression was significantly correlated with expression of active MMP-9 in ESCC tissues. Matrilysin-2 transfected TE-1 ESCC cells showed active MMP-9 activity and were more invasive in vitro compared with mock-transfected TE-1 cells. The results of this study suggest that matrilysin-2, the expression of which is closely correlated with nuclear beta-catenin expression and active MMP-9 activity, plays a key role in the progression of ESCC. PMID- 15333467 TI - Vitamin D receptor status alters mammary gland morphology and tumorigenesis in MMTV-neu mice. AB - The vitamin D(3) receptor (VDR) is a ligand-dependent transcription factor implicated in regulation of cell cycle, differentiation and apoptosis of both normal and transformed cells derived from mammary gland. In these studies we examined whether VDR status altered mammary gland morphology or transformation in the well-characterized MMTV-neu transgenic model of breast cancer. We demonstrate that VDR protein is highly expressed in neu-positive epithelial cells of preneoplastic lesions, established tumors and lung metastases from MMTV-neu mice. Furthermore, MMTV-neu mice lacking VDR exhibit abnormal mammary ductal morphology characterized by dilated, distended ducts containing dysplastic epithelial cells. From 12 months of age on, MMTV-neu mice lacking VDR also experience body weight loss, atrophy of the mammary fat pad, estrogen deficiency and reduced survival. The limited survival of MMTV-neu mice lacking VDR precluded an accurate assessment of the impact of complete VDR ablation on tumor development. MMTV-neu mice heterozygous for VDR, however, did not exhibit body weight loss, mammary gland atrophy or compromised survival. Compared with MMTV-neu mice with two copies of the VDR gene, haploinsufficiency of VDR shortened the latency and increased the incidence of mammary tumor formation. Tumor histology and expression/subcellular localization of the neu transgene were not altered by VDR haploinsufficiency despite a significant decrease in tumor VDR expression. Collectively, these studies suggest that VDR gene dosage impacts on age-related changes in ductal morphology and oncogene-induced tumorigenesis of the mammary gland in vivo. PMID- 15333468 TI - Acetyl-CoA carboxylase alpha gene and breast cancer susceptibility. AB - The identification of an interaction between BRCA1 and acetyl-CoA carboxylase alpha (ACCalpha), a key enzyme in lipid synthesis, led us to investigate the role of ACCalpha in breast cancer development, where it might contribute to the energy sensing mechanisms of malignant transformation. In order to investigate if certain ACCalpha alleles may be high-risk breast cancer susceptibility alleles, 37 extended breast and breast/ovarian cancer families negative for BRCA1 and BRCA2 mutations were exhaustively screened for sequence variations in the entire coding sequence, intron-exon junctions, 5'UTR, 3'UTR (untranslated regions) and the promoter regions of the ACCalpha gene. Two possibly disease-associated ACCalpha variants were each identified in a single family and were not present in 137 controls. Multiple polymorphisms were detected in breast cancer families, including 12 single nucleotide polymorphisms where the frequency of the rare allele estimated in controls was >0.10. The observed lack of variation in the ACCalpha coding region along with the presence of extended areas of linkage disequilibrium and low haplotype diversity indicates an overall high preservation of this gene. The prevalence of the ACCalpha haplotypes composed of common polymorphisms was determined in 453 breast cancer cases and 469 female controls. One haplotype was found to be associated with a substantial and highly significant increase in breast cancer risk (odds ratio = 3.10, 95% confidence interval 1.87-5.14, P < 0.0001), whereas three other haplotypes were found to have a protective effect. Our results indicate that mutations in the ACCalpha gene are unlikely to be a major cause of high-risk breast cancer susceptibility; however, certain common ACCalpha alleles may influence breast cancer risk. This study provides the first insight into the involvement of the ACCalpha gene in breast cancer predisposition and calls for further, large-scale studies that will be needed to understand the role of ACCalpha in tumour susceptibility and development. PMID- 15333469 TI - Haptoglobin phenotype and gestational diabetes. AB - OBJECTIVE: Haptoglobin (Hp), an Hb-binding plasma protein, exists in two major allelic variants. Hp1 has higher Hb binding and antioxidant capacity compared with Hp2. Individuals with Hp1 exhibit a lower incidence of angiopathies. Gestational diabetes mellitus (GDM) is an early manifestation of type 2 diabetes in pregnant women. It is usually confined to the time of gestation, but carries an increased risk to develop type 2 diabetes later in life. RESEARCH DESIGN AND METHODS: From consecutive Caucasian pregnant women (n = 250) referred for oral glucose tolerance testing, the Hp phenotype was determined. Significance of distribution and odds ratios (ORs) associated with Hp phenotype were calculated for women with GDM (n = 110) and women with normal glucose tolerance (n = 140). RESULTS: -Frequency of GDM in Hp phenotype classes increased with the number of Hp2 alleles (P < 0.001). ORs for GDM in women heterozygous and homozygous for Hp2 were 2.7 (95% CI 1.06-6.84) and 4.2 (1.67-10.55), respectively. CONCLUSIONS: Hp phenotype is an apparent risk factor for the development of GDM in our study population. This might be due to the low antioxidative potential of Hp2 compared with Hp1. PMID- 15333470 TI - A prospective study of red meat consumption and type 2 diabetes in middle-aged and elderly women: the women's health study. AB - OBJECTIVE: The aim of this study was to prospectively assess the relation between red meat intake and incidence of type 2 diabetes. RESEARCH DESIGN AND METHODS: Over an average of 8.8 years, we evaluated 37,309 participants in the Women's Health Study aged >/=45 years who were free of cardiovascular disease, cancer, and type 2 diabetes and completed validated semiquantitative food frequency questionnaires in 1993. RESULTS: During 326,876 person-years of follow-up, we documented 1,558 incident cases of type 2 diabetes. After adjusting for age, BMI, total energy intake, exercise, alcohol intake, cigarette smoking, and family history of diabetes, we found positive associations between intakes of red meat and processed meat and risk of type 2 diabetes. Comparing women in the highest quintile with those in the lowest quintile, the multivariate-adjusted relative risks (RRs) of type 2 diabetes were 1.28 for red meat (95% CI 1.07-1.53, P < 0.001 for trend) and 1.23 for processed meat intake (1.05-1.45, P = 0.001 for trend). Furthermore, the significantly increased diabetes risk appeared to be most pronounced for frequent consumption of total processed meat (RR 1.43, 95% CI 1.17-1.75 for >/=5/week vs. <1/month, P < 0.001 for trend) and two major subtypes, which were bacon (1.21, 1.06-1.39 for >/=2/week vs. <1/week, P = 0.004 for trend) and hot dogs (1.28, 1.09-1.50 for >/=2/week vs. <1/week, P = 0.003 for trend). These results remained significant after further adjustment for intakes of dietary fiber, magnesium, glycemic load, and total fat. Intakes of total cholesterol, animal protein, and heme iron were also significantly associated with a higher risk of type 2 diabetes. CONCLUSIONS: Our data indicate that higher consumption of total red meat, especially various processed meats, may increase risk of developing type 2 diabetes in women. PMID- 15333471 TI - Hepatitis C infection and type 2 diabetes in American-Indian women. AB - OBJECTIVE: The aim of this study was to describe the association between hepatitis C virus (HCV) infection and type 2 diabetes among a group of American Indian women who were screened for both conditions. RESEARCH DESIGN AND METHODS: The study population was a convenience sample of women who were receiving prenatal care. All women were systematically screened for both HCV and diabetes. RESULTS: A total of 426 women were included in the sample. HCV infection was detected in 13 (3.1% [95% CI 1.7-5.0]) and type 2 diabetes in 22 (5.2%, [3.3 7.6]) women. Women diagnosed with type 2 diabetes were more obese and had higher serum alanine aminotransferase activity compared with women without diabetes. Four of 13 (30.8% [10.6-58.7]) HCV-infected women and 18 of 413 (4.4% [2.7-6.7]) women without evidence of HCV infection had type 2 diabetes. (odds ratio 9.8 [95% CI 2.4-34.0], Fisher's exact test P = 0.003). In a logistic regression model, increasing age (10-year increments), obesity (by standard deviations from the mean BMI), and positive HCV status were each independently related to the diagnosis of diabetes. CONCLUSIONS: Among American-Indian women, type 2 diabetes is more common in those with than in those without HCV infection. This association and its potential mechanisms may have clinical implications. Investigation into the mechanisms linking HCV infection to the expression of type 2 diabetes may also help to define processes that promote the development of type 2 diabetes in susceptible individuals. PMID- 15333472 TI - Cost-effectiveness analysis of different screening procedures for type 2 diabetes: the KORA Survey 2000. AB - OBJECTIVE: To compare the cost-effectiveness of different type 2 diabetes screening strategies using population-based data (KORA Survey; Augsburg, Germany; subjects aged 55-74 years), including participation data. RESEARCH DESIGN AND METHODS: The decision analytic model, which had a time horizon of 1 year, used the following screening strategies: fasting glucose testing, the oral glucose tolerance test (OGTT) following fasting glucose testing in impaired fasting glucose (IFG) (fasting glucose + OGTT), OGTT only, and OGTT if HbA(1c) was >5.6% (HbA(1c) + OGTT), all with or without first-step preselection (p). The main outcome measures were costs (in Euros), true-positive type 2 diabetic cases, incremental cost-effectiveness ratios (ICERs), third-party payers, and societal perspectives. RESULTS: After dominated strategies were excluded, the OGTT and HbA(1c) + OGTT from the perspective of the statutory health insurance remained, as did fasting glucose + OGTT and HbA(1c) + OGTT from the societal perspective. OGTTs (4.90 per patient) yielded the lowest costs from the perspective of the statutory health insurance and fasting glucose + OGTT (10.85) from the societal perspective. HbA(1c) + OGTT was the most expensive (21.44 and 31.77) but also the most effective (54% detected cases). ICERs, compared with the next less effective strategies, were 771 from the statutory health insurance and 831 from the societal perspective. In the Monte Carlo analysis, dominance relations remained unchanged in 100 and 68% (statutory health insurance and societal perspective, respectively) of simulated populations. CONCLUSIONS: The most effective screening strategy was HbA(1c) combined with OGTT because of high participation. However, costs were lower when screening with fasting glucose tests combined with OGTT or OGTT alone. The decision regarding which is the most favorable strategy depends on whether the goal is to identify a high number of cases or to incur lower costs at reasonable effectiveness. PMID- 15333473 TI - Costs of lower-extremity ulcers among patients with diabetes. AB - OBJECTIVE: The objective of this study was to characterize health care costs associated with diabetic lower-extremity ulcers. RESEARCH DESIGN AND METHODS: Adult patients with diabetes who had a lower-extremity ulcer episode during 2000 and 2001 were identified using claims data. Ulcer-related direct health care costs were computed for each episode. Episodes were stratified according to severity level based on the Wagner classification. RESULTS: A total of 2,253 patients were identified. The mean age was 68.9 years, and 59% of the patients were male. The average episode duration was 87.3 +/- 82.8 days. Total ulcer related costs averaged 13,179 dollars per episode and increased with severity level, ranging from 1,892 dollars (level 1) to 27,721 dollars (level 4/5). Inpatient hospital charges accounted for 77% (10,188 dollars) of the overall cost, indicating that hospitalization was a major cost driver. Total ulcer related costs were significantly higher for patients <65 years of age compared with those of older patients (16,390 dollars vs. 11,925 dollars, P = 0.02) and for patients with inadequate vascular status compared with patients with adequate vascular status (23,372 dollars vs. 5,218 dollars, P < 0.0001). Patients who progressed to a higher severity level also had significantly higher ulcer-related costs compared with patients who did not progress (20,136 dollars vs. 3,063 dollars, P < 0.0001). CONCLUSIONS: The high costs of treating diabetic lower extremity ulcers emphasize the value of intensive outpatient interventions designed to prevent ulcer progression. PMID- 15333474 TI - Prevalence of the metabolic syndrome and its components: findings from a Finnish general population sample and the Diabetes Prevention Study cohort. AB - OBJECTIVE: To assess the prevalence of the metabolic syndrome (MetS) in two independent Finnish study cohorts. RESEARCH DESIGN AND METHODS: The prevalence of the MetS by modified World Health Organization criteria was analyzed in different categories of glucose tolerance in a cross-sectional, population-based sample of 2,049 individuals (FINRISK) aged 45-64 years and in 522 participants of the Finnish Diabetes Prevention Study (DPS) with impaired glucose tolerance (IGT). RESULTS: In the FINRISK cohort, the MetS was present in 38.8% of the men and 22.2% of the women. The prevalence was 14.4 and 10.1% in subjects with normal glucose tolerance, 74.0 and 52.2% in subjects with impaired fasting glucose, 84.8 and 65.4% in subjects with IGT, and 91.5 and 82.7% in subjects with type 2 diabetes in men and women, respectively. Among women, the prevalence of the MetS increased with increasing age. In the DPS cohort, the MetS was present in 78.4% of the men and 72.2% of the women with IGT. CONCLUSIONS: The MetS was extremely common in middle-aged subjects The high prevalence in men was mostly due to their high waist-to-hip ratio. The prevalence of the MetS increased in both sexes with deterioration in glucose regulation. Approximately 75% of the subjects with IGT had the MetS. Because the syndrome includes the major risk factors for atherosclerotic vascular diseases and is the major antecedent for type 2 diabetes, concerted preventive action should be targeted to control all the features of the MetS. PMID- 15333475 TI - Features of the metabolic syndrome are associated with objectively measured physical activity and fitness in Danish children: the European Youth Heart Study (EYHS). AB - OBJECTIVE: Features of the metabolic syndrome are becoming increasingly evident in children. Decreased physical activity is likely to be an important etiological factor, as shown previously for subjective measures of physical activity in selected groups. The purpose of this study was to examine the relationship between the metabolic syndrome and objectively measured physical activity and whether fitness modified this relationship. RESEARCH DESIGN AND METHODS: A total of 589 Danish children (310 girls, 279 boys, mean [+/-SD] age 9.6 +/- 0.44 years, mean weight 33.6 +/- 6.4 kg, mean height 1.39 +/- 0.06 m) were randomly selected. Physical activity was measured with the uni-axial Computer Science & Applications accelerometer (MTI actigraph) worn at the hip for at least 3 days (>/=10 h/day) and fitness with a maximal bike test. As outcomes, we measured sitting systolic and diastolic blood pressure, degree of adiposity (sum of four skinfolds), and, finally, insulin, glucose, triglicerides, and HDL cholesterol in fasting blood samples. The outcome variables were statistically normalized and expressed as the number of SDs from the mean. (i.e., Z scores). A metabolic syndrome risk score was computed as the mean of these Z scores. Multiple linear regression was used to test the association between physical activity and metabolic risk, adjusted primarily for age, sex, sexual maturation, ethnicity, parental smoking, socioeconomic factors, and the Computer Science & Applications unit, as well as for fitness. Robust SEs were computed by clustering on school. RESULTS: All children were in the nondiabetic range of fasting glucose. Metabolic risk was inversely related to physical activity (P = 0.008). The relationship was weakened after adjustment for fitness, but there was a significantly positive interaction between physical activity and fitness. CONCLUSIONS: Physical activity is inversely associated with metabolic risk, independently of potential confounders. The interaction between physical activity and fitness suggests that the potential beneficial effect of activity may be greatest in children with lower cardiorespiratory fitness. PMID- 15333476 TI - Oral antihyperglycemic medication nonadherence and subsequent hospitalization among individuals with type 2 diabetes. AB - OBJECTIVE: This study examines the association between oral antihyperglycemic medication nonadherence and subsequent hospitalization among individuals with type 2 diabetes. RESEARCH DESIGN AND METHODS: Using administrative claims data (2000-2001) from a managed care organization in the Midwestern U.S., this study analyzed 900 enrollees, aged 18 years and over, with type 2 diabetes who were taking oral antihyperglycemic agents both years but who did not use insulin. Nonadherence was defined as a medication possession ratio (MPR) <80%. Multivariate logistic regression analyses were performed where hospitalization in 2001 was regressed on nonadherence to the oral antihyperglycemic drug regimen in 2000, while controlling for nonadherence to drugs for hypertension and dyslipidemia and for hospitalization in 2000, age, sex, intensity of the diabetes drug regimen, and comorbidities. RESULTS: The proportion of enrollees who were nonadherent to the antihyperglycemic drug regimen in 2001 was 28.9%, whereas 18.8 and 26.9% were nonadherent to antihypertensive and lipid-modifying drugs, respectively. The increase in the hospitalization rate for 2001 was most apparent where the antihyperglycemic MPR for 2000 dropped to <80%. Enrollees who were nonadherent to oral diabetes medications in 2000 were at higher risk of hospitalization in 2001 (odds ratio 2.53; 95% CI 1.38-4.64), whereas nonadherence to drugs for hypertension and dyslipidemia were not significantly associated with hospitalization. CONCLUSIONS: Patients with type 2 diabetes who do not obtain at least 80% of their oral antihyperglycemic medications across 1 year are at a higher risk of hospitalization in the following year. PMID- 15333477 TI - Relationship of depression and diabetes self-care, medication adherence, and preventive care. AB - OBJECTIVE: We assessed whether diabetes self-care, medication adherence, and use of preventive services were associated with depressive illness. RESEARCH DESIGN AND METHODS: In a large health maintenance organization, 4,463 patients with diabetes completed a questionnaire assessing self-care, diabetes monitoring, and depression. Automated diagnostic, laboratory, and pharmacy data were used to assess glycemic control, medication adherence, and preventive services. RESULTS: This predominantly type 2 diabetic population had a mean HbA(1c) level of 7.8 +/- 1.6%. Three-quarters of the patients received hypoglycemic agents (oral or insulin) and reported at least weekly self-monitoring of glucose and foot checks. The mean number of HbA(1c) tests was 2.2 +/- 1.3 per year and was only slightly higher among patients with poorly controlled diabetes. Almost one-half (48.9%) had a BMI >30 kg/m(2), and 47.8% of patients exercised once a week or less. Pharmacy refill data showed a 19.5% nonadherence rate to oral hypoglycemic medicines (mean 67.4 +/- 74.1 days) in the prior year. Major depression was associated with less physical activity, unhealthy diet, and lower adherence to oral hypoglycemic, antihypertensive, and lipid-lowering medications. In contrast, preventive care of diabetes, including home-glucose tests, foot checks, screening for microalbuminuria, and retinopathy was similar among depressed and nondepressed patients. CONCLUSIONS: In a primary care population, diabetes self care was suboptimal across a continuum from home-based activities, such as healthy eating, exercise, and medication adherence, to use of preventive care. Major depression was mainly associated with patient-initiated behaviors that are difficult to maintain (e.g., exercise, diet, medication adherence) but not with preventive services for diabetes. PMID- 15333478 TI - Rates of glucose change measured by blood glucose meter and the GlucoWatch Biographer during day, night, and around mealtimes. AB - OBJECTIVE: The purpose of this study was to characterize the distribution of the rate of change of blood glucose for a diabetic population. RESEARCH DESIGN AND METHODS: The study population consisted of 124 adults with type 1 or type 2 diabetes requiring insulin. Study participants applied a GlucoWatch Biographer during the day at home for 5 consecutive days and took finger-prick blood glucose measurements hourly. Subjects kept a diary of meals. The Biographer frequently and automatically measured glucose up to three times per hour for up 12 h. Rates of glucose change were calculated for both Biographer and blood glucose measurements. Rates of glucose change during a separate study of 134 subjects were determined for daytime and nighttime use. RESULTS: -Mean (+/-SD) rates of change of glucose of -0.36 +/- 0.95 and 0.36 +/- 0.99 mg. dl(-1). min(-1) were found before and after lunch using blood glucose data and -0.31 +/- 1.23 and 0.43 +/- 1.26 using Biographer data. For both types of diabetes, rates of glucose change exceeded 2 mg. dl(-1). min(-1) before and after meals approximately 10% of the time. Periprandial glucose patterns showed some significant differences between type 1 and type 2 diabetic subjects. Glucose levels changed more gradually at night than during the day. CONCLUSIONS: Glucose values were almost equally unstable before and after meals. Glycemic instability around dinner was different in type 1 and type 2 diabetes. The GlucoWatch Biographer was found to be effective in tracking trends in glucose levels and yielded similar results as obtained by finger-prick blood samples. PMID- 15333479 TI - Effect of high-dose vitamin E on insulin resistance and associated parameters in overweight subjects. AB - OBJECTIVE: Markers of oxidative stress and plasma alanine transferase (ALT) levels are increased and circulating antioxidant concentrations are reduced in individuals with insulin resistance. Vitamin E improves glycemic control in people with diabetes. We tested the hypothesis that vitamin E would decrease markers of oxidative stress and plasma ALT levels and improve insulin sensitivity in overweight individuals. RESEARCH DESIGN AND METHODS: Eighty overweight individuals (BMI >27 kg/m(2)) were randomly allocated to receive either 800 IU vitamin E per day or a matching placebo for 3 months. The dose of vitamin E was increased to 1,200 IU per day for a further 3 months. RESULTS: Plasma peroxides decreased by 27% at 3 months and by 29% at 6 months in the group that received vitamin E and were positively correlated with plasma vitamin E concentrations at the 6-month time point. At 3 months, fasting plasma glucose and insulin concentrations were significantly reduced and homeostasis model assessment increased. These changes were not apparent at 6 months. Plasma ALT concentrations declined significantly throughout the study period. CONCLUSIONS: In conclusion, these findings indicate that vitamin E improves oxidative stress and hepatocellular function. Although insulin resistance also improves, this effect appears transient. PMID- 15333480 TI - Effect of training status on regional disposal of circulating free fatty acids in the liver and skeletal muscle during physiological hyperinsulinemia. AB - OBJECTIVE: Fat metabolism is increasingly implicated in the pathogenesis of type 2 diabetes. Endurance training has been shown to prevent hepatic steatosis and to alter skeletal muscle fat metabolism, and regional free fatty acid (FFA) uptake adaptations were suggested as a mechanism. Thus, we tested whether endurance training modifies the uptake of plasma FFAs occurring in the liver and in skeletal muscle during anabolic, i.e., hyperinsulinemic, conditions. RESEARCH DESIGN AND METHODS: Trained and untrained healthy male subjects underwent positron emission tomography scanning of the liver and thigh regions, with the FFA analog 14(R,S)-[(18)F]fluoro-6-thia-heptadecanoic acid, during euglycemic hyperinsulinemia. Tracer influx rate constants in skeletal muscle (MK(i)) and liver (LK(i)) were multiplied by plasma FFA levels to obtain FFA uptake for skeletal muscle (MFU) and liver (LFU), respectively. RESULTS: Athletes showed increased Vo(2max) (P < 0.0001), insulin-mediated glucose disposal (M value, 61 +/- 4 vs. 46 +/- 3 micromol. min(-1). kg(-1), P = 0.01), and plasma lactate levels during the clamp and lower percentage of body fat mass (P = 0.002). MK(i) was 25% higher in athletes than in sedentary men (P = 0.03). In all subjects, MK(i) and MFU were positively correlated with the M value (r = 0.56, P = 0.02, and r = 0.51, P = 0.03, respectively) and with plasma lactate levels (r = 0.63, P = 0.006, and r = 0.63, P = 0.005, respectively). LK(i) was significantly reduced by 20% in the athletes (P = 0.04). By multiple regression, LFU was inversely correlated with the two fitness categories (P = 0.008), and it was lower in athletes. Linear fitting of liver data showed time consistency, indicating no release of FFAs as a mechanism for the reduced liver retention in athletes. CONCLUSIONS: We conclude that endurance training promotes insulin-mediated glucose and FFA disposal in skeletal muscle, while lowering hepatic FFA uptake. Such changes may result in a divergent pattern of fat accumulation in the two organs. PMID- 15333481 TI - Oxidative stress and antioxidant defense in relation to the severity of diabetic polyneuropathy and cardiovascular autonomic neuropathy. AB - OBJECTIVE: Oxidative stress resulting from enhanced free-radical formation and/or a defect in antioxidant defenses has been implicated in the pathogenesis of experimental diabetic neuropathy. The objective of this study was to evaluate plasma levels of various biomarkers of oxidative stress in diabetic subjects in relation to the presence or absence of polyneuropathy (PN) and/or cardiovascular autonomic neuropathy (CAN). RESEARCH DESIGN AND METHODS: Plasma 8-iso prostaglandin F(2alpha) (8-iso-PGF(2alpha)), superoxide anion (O(2)(.-)) generation, lag phase to peroxidation by peroxynitrite (ONOO(-)), vitamin E-to lipid ratio, and vitamin C were measured in nonsmoking diabetic patients without PN and CAN (PN(-)/CAN(-) group; n = 62), in a group with PN but without CAN (PN(+)/CAN(-) group; n = 105), in those with both PN and CAN (PN(+)/CAN(+) group; n = 22), and in healthy control subjects (n = 85). RESULTS: All three markers of oxidative stress were significantly increased, and both markers of antioxidant defense were decreased in the PN(+)/CAN(-) group compared with the control group (all P < 0.05). PN(-)/CAN(-) subjects showed a significant increase compared with control subjects for 8-iso-PGF(2alpha), O(2)(.-), and ONOO(-) and a decrease for the vitamin E-to-lipid ratio (all P < 0.05). In the PN(+)/CAN(-) group, a significant increase compared with the PN(-)/CAN(-) group was noted for O(2)(.-), whereas the vitamin E-to-lipid ratio and vitamin C were reduced (all P < 0.05). No significant differences were noted between the PN(+)/CAN(-) and PN(+)/CAN(+) groups for each of the five markers of oxidative stress. In multivariate models, O(2)(.-) and ONOO(-) were independently associated with neuropathic deficits, but diabetes duration and triglyceride levels were also independent determinants. CONCLUSIONS: Oxidative stress is enhanced in diabetic patients before the development of PN but to an even higher degree in those with PN, without further significant increase in relation to superimposed autonomic neuropathy. However, apart from oxidative stress, diabetes duration and triglyceride levels are also related to the severity of PN. PMID- 15333482 TI - Endothelial nitric oxide synthase gene is associated with diabetic macular edema in type 2 diabetes. AB - OBJECTIVE: We examined the endothelial nitric oxide (eNOS) gene polymorphisms to assess its possible association with diabetic retinopathy and macular edema. RESEARCH DESIGN AND METHODS: A total of 226 patients with type 2 diabetes and 186 healthy subjects were studied. Type 2 diabetic patients consisted of 110 patients without retinopathy, 46 patients with nonproliferative diabetic retinopathy, and 71 patients with proliferative diabetic retinopathy. Diabetic macular edema was present in 48 patients. Three polymorphisms of the eNOS gene were determined: T 786C in the promoter region, 27-bp repeat in intron 4, and Glu298Asp in exon 7. RESULTS: Close linkage disequilibrium was observed between the T-786C polymorphism and the 27-bp repeat, as has been previously reported, but Glu298Asp was not in linkage disequilibrium with the other two polymorphisms. The eNOS gene polymorphisms were not significantly associated with the presence of retinopathy or with retinopathy severity or type 2 diabetes itself. However, by both association study and multiple logistic regression analysis, the T-786C and 27-bp repeat polymorphisms were significantly associated with a risk of developing macular edema with the -786C allele and the "a" allele increasing the risk. CONCLUSIONS: The present study suggests that the eNOS gene is a novel genetic risk factor for diabetic macular edema. The eNOS gene polymorphisms may contribute to the development of macular edema by impairing basal eNOS expression and resulting in the breakdown of the blood-retina barrier. PMID- 15333483 TI - Temporary preservation of beta-cell function by diazoxide treatment in childhood type 1 diabetes. AB - OBJECTIVE: We examined the effect of diazoxide, an ATP-sensitive K(+) channel opener and inhibitor of insulin secretion, on beta-cell function and remission in children at clinical onset of type 1 diabetes. RESEARCH DESIGN AND METHODS: A total of 56 subjects (21 girls and 35 boys, age 7-17 years) were randomized to 3 months of active treatment (diazoxide 5-7.5 mg/kg in divided doses) or placebo in addition to multiple daily insulin injections and were followed for 2 years. RESULTS: Diazoxide decreased circulating C-peptide concentrations by approximately 50%. After cessation of the treatment, basal and meal-stimulated C peptide concentrations increased to a maximum at 6 months, followed by a decline. Meal-stimulated C-peptide concentration was significantly higher at 12 months (0.43 +/- 0.22 vs. 0.31 +/- 0.26 nmol/l, P = 0.018) and tended to fall less from clinical onset to 24 months in the diazoxide- vs. placebo-treated patients (-0.05 +/- 0.24 vs. -0.18 +/- 0.26 nmol/l, P = 0.064). At 24 months, the meal-stimulated C-peptide concentrations were 0.24 +/- 0.20 and 0.20 +/- 0.17 nmol/l, respectively. Side effects of diazoxide were prevalent. CONCLUSIONS: This study demonstrates that partial inhibition of insulin secretion for 3 months at onset of childhood type 1 diabetes suspends the period of remission and temporarily preserves residual insulin production. Further evaluation of the full potential of beta-cell rest will require compounds with less side effects as well as protocols optimized for sustained secretory arrest. PMID- 15333484 TI - Diabetes is the strongest risk factor for lower-extremity amputation in new hemodialysis patients. AB - OBJECTIVE: End-stage renal disease (ESRD) patients, especially those with diabetes, have an increased risk of nontraumatic lower-extremity amputation (LEA). The present study aims to examine the association of demographic and clinical variables with the risk of hospitalization for LEA among incident hemodialysis patients. RESEARCH DESIGN AND METHODS: The study population consisted of incident hemodialysis patients from the study years 1996-1999 of the ESRD Core Indicator/Clinical Performance Measures (CPM) Project. Cox proportional hazard modeling was used to identify factors associated with LEA. RESULTS: Four percent (116 of 3,272) of noncensored incident patients had an LEA during the 12 month follow-up period. Factors associated with LEA included diabetes as the cause of ESRD or preexisting comorbidity (hazard ratio 6.4, 95% CI 3.4-12.0), cardiovascular comorbidity (1.8, 1.2-2.8), hemodialysis inadequacy (urea reduction ratio [URR] <58.5% (1.9, 1.1-3.3), and lower serum albumin level (1.6, 1.1-2.3). Among patients with diabetes, hemodialysis inadequacy and cardiovascular comorbidity were risk factors for LEA (2.6, 1.4-4.8, and 1.7, 1.1 2.6, respectively). CONCLUSIONS: These data suggest that diabetes is a potent risk factor for LEA in new hemodialysis patients. In ESRD patients with diabetes, a multipronged approach may reduce the rate of LEA. Potentially beneficial strategies include adherence to hemodialysis adequacy guidelines, aggressive treatment of cardiovascular comorbidities, and the utilization of LEA prevention strategies recommended for the general population of patients with diabetes. PMID- 15333485 TI - Contribution of insulin-stimulated glucose uptake and basal hepatic insulin sensitivity to surrogate measures of insulin sensitivity. AB - OBJECTIVE: The goal of this study was to evaluate the performance of surrogate measures of insulin sensitivity and insulin secretion. RESEARCH DESIGN AND METHODS: The homeostasis model assessment (HOMA) of insulin resistance (IR) and the insulin sensitivity index (S(i)) from oral glucose tolerance test (OGTT) were compared with the M value from a hyperinsulinemic-euglycemic clamp in 467 subjects with various degrees of glucose tolerance. Endogenous glucose production (EGP) and hepatic insulin sensitivity were determined in a subset (n = 143). Insulin secretion was estimated as the HOMA beta-cell index and as the insulinogenic index from the first 30 min of the OGTT (I/G30) and compared with the first-phase insulin response (FPIR) to an intravenous glucose tolerance test (n = 218). RESULTS: The M value correlated with the HOMA-IR (r = -0.591, P < 0.0001) and the S(i) (r = 0.533, P < 0.0001) indexes in the total study group. HOMA-IR correlated with basal EGP in the total study group (r = 0.378, P < 0.0005) and in subjects with diabetes (r = 0.330, P = 0.01). However, neither HOMA-IR nor S(i) correlated significantly with the M value in subjects with impaired fasting glucose (IFG) (r = -0.108, P = 0.5; r = 0.01, P = 0.9) or IFG/impaired glucose tolerance (IGT) (r = -0.167, P = 0.4; r = 0.09, P = 0.6). The HOMA-IR correlated with hepatic insulin sensitivity in the whole study group (r = -0.395, P < 0.005) as well as in the IFG/IGT subgroup (r = -0.634, P = 0.002) and in the diabetic subgroup (r = -0.348, P = 0.008). In subjects with IFG/IGT, hepatic insulin sensitivity was the most important determinant of HOMA IR, explaining 40% of its variation. The HOMA beta-cell index showed a weak correlation with FPIR in the whole study group (r = 0.294, P = 0.001) but not in the subgroups. In contrast, the I/G30 correlated with FPIR in the whole study group (r = 0.472, P < 0.0005) and in the IFG/IGT subgroup (r = 0.493, P < 0.005). CONCLUSIONS: HOMA-IR is dependent upon both peripheral and hepatic insulin sensitivity, the contribution of which differs between subjects with normal and elevated fasting glucose concentrations. These discrepancies develop as a consequence of a nonparallel deterioration of the variables included in the equations with worsening of glucose tolerance. PMID- 15333486 TI - Lower toenail chromium in men with diabetes and cardiovascular disease compared with healthy men. AB - OBJECTIVE: Chromium may improve insulin sensitivity, which can modify the risk of diabetes and cardiovascular disease (CVD). Therefore, we evaluated the association between toenail chromium and CVD in diabetic men. RESEARCH DESIGN AND METHODS: We performed cross-sectional and nested case-control analyses among men aged 40-75 years within the Health Professionals Follow-up Study. The cross sectional analysis compared men with diabetes only (n = 688), diabetes with prevalent CVD (n = 198), and healthy control subjects (n = 361). The nested case control study included 202 men with baseline diabetes who developed incident CVD and 361 matched control subjects. RESULTS: Mean toenail chromium (microg/g) was 0.71 in healthy control subjects, 0.61 in diabetes-only subjects, and 0.52 in diabetic subjects with prevalent CVD (P for trend = 0.003). In the cross sectional analysis, the multivariate odds ratio (OR) between extreme quartiles was 0.74 (95% CI 0.49-1.11; P for trend = 0.18), comparing diabetes only with healthy control subjects. A similar comparison between diabetic subjects with prevalent CVD and healthy control subjects yielded an OR of 0.45 (0.24-0.84; P for trend = 0.003). In the nested case-control study, comparing diabetic men with incident CVD with healthy control subjects, the multivariate OR was 0.65 (0.36 1.17; P for trend = 0.16) between extreme quartiles. When we combined prevalent and incident CVD cases among diabetic men and compared them with healthy control subjects, the OR was 0.62 (0.39-1.01; P for trend = 0.02) between extreme quartiles. CONCLUSIONS: Our results suggest that diabetic men with CVD have lower toenail chromium than healthy control subjects. However, this study could not distinguish between the effects of chromium on diabetes and those on CVD. Long term clinical trials are needed to determine whether chromium supplementation is beneficial for preventing CVD among diabetic patients. PMID- 15333487 TI - Possible impairment of transcardiac utilization of adiponectin in patients with type 2 diabetes. AB - OBJECTIVE: Adiponectin, an adipocyte-derived protein, has been suggested to enhance insulin sensitivity and prevent atherosclerosis. Circulating adiponecin levels are reduced in states of insulin resistance such as type 2 diabetes. We examined transcardiac utilization of adiponectin in patients with and without type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 17 male type 2 diabetic patients and 17 male nondiabetic patients were investigated. Venous blood samples were taken to measure glucose and lipid variables. Blood samples for the measurement of adiponectin were collected simultaneously from the aortic root and coronary sinus. Angiographic semiquantitative stenosis score of coronary artery was also evaluated. RESULTS: The adiponectin levels in both the aortic root and coronary sinus in the diabetic patients were significantly lower than those in the nondiabetic patients. The adiponectin level was significantly lower in the coronary sinus than in the aortic root in the nondiabetic patients, but there was no significant difference between adiponectin levels in the aortic root and coronary sinus in the diabetic patients. The total stenosis score, as an index of severity of coronary artery stenosis, was significantly higher in the diabetic patients than in the nondiabetic patients. The stenosis score was correlated with the degree of transcardiac utilization of adiponectin from the aortic root to coronary sinus in the nondiabetic patients but not in the diabetic patients. CONCLUSIONS: Diabetic patients not only have a decreased adiponectin level in the basal state compared with nondiabetic patients but also have impaired utilization of adiponectin in the coronary artery and/or the heart, which may promote the development of atherosclerosis. PMID- 15333488 TI - Metabolic syndrome in normal-weight Americans: new definition of the metabolically obese, normal-weight individual. AB - OBJECTIVE: To determine the prevalence rates and likelihood of the metabolic syndrome and its individual components in normal-weight and slightly overweight individuals (BMI 18.5-26.9 kg/m(2)). RESEARCH DESIGN AND METHODS: There were a total of 7,602 adult participants of the Third National Health and Nutrition Examination Survey, a nationally representative cross-sectional survey. Prevalence and odds ratios (ORs) of the metabolic syndrome, defined according to National Cholesterol Education Program Adult Treatment Panel III criteria, were computed according to 2.0- to 2.5-unit increments in BMI. RESULTS: Depending on ethnicity and sex, the prevalence of the metabolic syndrome increased in a graded fashion from 0.9-3.0% at BMI 18.5-20.9 kg/m(2) to 9.6-22.5% at BMI 25.0-26.9 kg/m(2). Compared with men with BMI 18.5-20.9 kg/m(2), the odds for the metabolic syndrome were 4.13 (95% CI 1.57-10.87) for men with BMI 21-22.9 kg/m(2), 5.35 (2.41-11.86) for men with BMI 23-24.9 kg/m(2), and 9.08 (4.23-19.52) for men with BMI 25-26.9 kg/m(2) after controlling for age, ethnicity, education, income, physical activity, smoking status, and alcohol and total fat, saturated fat, carbohydrate, and fiber intakes. The corresponding ORs in women were 4.34 (2.08 9.07), 7.77 (3.95-15.26), and 17.34 (9.29-32.38). CONCLUSIONS: Individuals in the upper normal-weight and slightly overweight BMI range have a relatively high prevalence and are at increased risk of having the metabolic syndrome. Therefore, screening in individuals with normal or slightly elevated BMI is important in the prevention of diabetes and cardiovascular disease. PMID- 15333489 TI - Lack of agreement between the revised criteria of impaired fasting glucose and impaired glucose tolerance in children with excess body weight. AB - OBJECTIVE: The aim of this study was to describe the agreement between impaired glucose tolerance (IGT) and impaired fasting glucose (IFG) in children with excess body weight using the original and the revised definitions of IFG. RESEARCH DESIGN AND METHODS: Obese and overweight children aged 4-17 years were included (n = 533). Anthropometric parameters and biochemical tests (fasting and 2-h glucose tests after an oral glucose load [1.75 g/kg]) were performed. Case subjects with a fasting plasma glucose >/=126 mg/dl were excluded. The diagnostic parameters of the original and the revised definitions of IFG for detecting IGT were estimated. The analysis of agreement between these categories was made using the kappa test. RESULTS: The prevalence of IFG increased from 6.2 to 13.3% using the new criteria. The prevalence of IFG became closer to the prevalence of IGT (14.8%). The revised criteria increased the sensitivity from 26.6 to 36.7%. However, the new IFG definition was not useful for identifying IGT cases. Of the 71 case subjects with IFG, only 29 (40.8%) had IGT. In addition, 50 case subjects with IGT (9.4%) and 13 with diabetes (2.4%) had a fasting glycemia <100 mg/dl. A poor agreement was found between the 2003 IFG definition and abnormal 2-h postchallenge plasma glucose (kappa = 0.359). The proportion of false-positive cases increased (36.3-59.1%) under the new definition. CONCLUSIONS: The new definition modestly increases the sensitivity of IFG for detecting IGT in children with excess body weight. Despite this, more than one-half of these cases are not detected. In addition, the false-positive rate was increased by 61%. PMID- 15333490 TI - Cardiovascular disease risk factors predict the development of type 2 diabetes: the insulin resistance atherosclerosis study. AB - OBJECTIVE: In a few previous studies, cardiovascular disease (CVD) risk factors (RFs) have been shown to predict diabetes. Our objective was to determine whether the presence of CVD RFs predict the eventual development of diabetes after controlling for known RFs, such as directly measured insulin resistance and obesity. RESEARCH DESIGN AND METHODS: We studied 872 participants with normal or impaired glucose tolerance (IGT) who were enrolled at baseline in the Insulin Resistance Atherosclerosis Study (IRAS). Of these, 143 (16%) developed type 2 diabetes in 5 years. Using these participants, a series of logistic regression models were fit to address the question. RESULTS: Significant RFs for developing type 2 diabetes included high plasminogen activator inhibitor-1, hypertension, high triglycerides, low levels of HDL cholesterol, and IGT. The 5-year cumulative incidence of type 2 diabetes by the number of RFs (0-5) was as follows: no RFs, 11 of 230 = 5%; one RF, 31 of 278 = 11%; two RFs, 36 of 202 = 18%; three RFs, 41 of 110 = 37%; four RFs, 19 of 42 = 45%; and five RFs, 5 of 10 = 50% (P < 0.001). The odds ratio (OR) for conversion to type 2 diabetes for each additional RF was 2.1 (95% CI 1.78-2.46) after adjusting for age, sex, ethnicity, and center. After further adjustment for insulin resistance, determined by the frequently sampled intravenous glucose tolerance test and waist circumference, each additional CVD RF increased the risk of type 2 diabetes significantly (OR 1.81, 95% CI 1.49 2.20). CONCLUSIONS: Individuals with multiple CVD RFs are at increased risk of type 2 diabetes, which is only partially mediated by insulin resistance or central adiposity. This information should be useful for identifying high-risk patients for developing diabetes through RF assessments. PMID- 15333491 TI - Combination therapy with fenofibrate and rosiglitazone paradoxically lowers serum HDL cholesterol. PMID- 15333492 TI - Beta-cell transplantation restores metabolic control and quality of life in a patient with subcutaneous insulin resistance. PMID- 15333493 TI - Hematocrit and the incidence of type 2 diabetes in the pima indians. PMID- 15333494 TI - The impact of family history of diabetes on glucose testing and counseling behavior in primary care. PMID- 15333495 TI - Disease management advice provided to African-American and Chinese-American patients with type 2 diabetes. PMID- 15333496 TI - High incidence of type 2 diabetes in peroxisome proliferator-activated receptor gamma2 Pro12Ala carriers exposed to a high chronic intake of trans fatty acids and saturated fatty acids. PMID- 15333497 TI - Lipids and glucose in type 2 diabetes: what is the cause and effect? PMID- 15333498 TI - Indirect support for the use of supplemental insulin in hospitalized insulin requiring diabetic patients. PMID- 15333499 TI - Guidelines for computer modeling of diabetes and its complications. PMID- 15333500 TI - Dietary carbohydrate (amount and type) in the prevention and management of diabetes: a statement by the american diabetes association. PMID- 15333501 TI - Inpatient diabetes control: approaches to treatment. PMID- 15333502 TI - Prevalence of eating disorders in young patients with type 1 diabetes from two different Italian cities. PMID- 15333503 TI - Hypoglycemic coma in a diabetic patient on peritoneal dialysis due to interference of icodextrin metabolites with capillary blood glucose measurements. PMID- 15333504 TI - Resolution of diabetic cheiroarthropathy after pancreatic transplantation. PMID- 15333505 TI - The Korle-Bu Hb variant in Caucasian women with type 1 diabetes: a pitfall in the assessment of diabetes control. PMID- 15333506 TI - Short-term intensive insulin therapy in newly diagnosed type 2 diabetes: response to Ryan, Imes, and Wallace. PMID- 15333508 TI - Deficiency in the detection of microalbuminuria by urinary dipstick in diabetic patients: response to Comper et al. PMID- 15333510 TI - A systematic review of adherence with medications for diabetes: response to Cramer. PMID- 15333512 TI - The GILHT-E Syndrome? PMID- 15333513 TI - Effects of avasimibe on cytochrome P450 2C9 expression in vitro and in vivo. AB - Avasimibe, an acyl-CoA:cholesterol acyltransferase inhibitor, has been previously shown to be a potent inducer of CYP3A4 and multiple drug resistance protein 1. We have further characterized the drug interaction potential of avasimibe by studying the inductive and inhibitory effect of this compound on major drug metabolizing enzymes. Enzymes known to be involved in the metabolism of drugs likely to be coadministered with avasimibe, such as CYP1A1/2, CYP2C, and CYP2B6, were evaluated further by microarray analysis, Western immunoblotting, and activity assays, using rifampicin and beta-naphthoflavone as positive controls. No change was observed in CYP1A1/2 mRNA or activity levels after avasimibe treatment. Differential induction of CYP2C9- and CYP2B6-immunoreactive protein and activity was observed depending on drug concentration and donor. Microarray analysis showed a similar increase in CYP2C and CYP2B6 mRNA levels. The inhibition potential of avasimibe on the major drug-metabolizing enzymes was assessed using pooled human liver microsomes. Avasimibe inhibited CYP2C9 (IC50 2.9 microM), CYP1A2 (IC50 13.9 microM), and CYP2C19 (IC50 26.5 microM). A clinical drug interaction study was conducted to determine whether avasimibe might interact with the CYP2C9 substrate warfarin. Volunteers received 750 mg of avasimibe and showed a 54.2% reduction in trough concentrations of S-warfarin and decreased prothrombin times by 12, 15, 19, and 21% on days 6 through 9, respectively. These results demonstrate that avasimibe's inductive spectrum resembles that of rifampin. PMID- 15333514 TI - High absorption but very low bioavailability of oral resveratrol in humans. AB - The dietary polyphenol resveratrol has been shown to have chemopreventive activity against cardiovascular disease and a variety of cancers in model systems, but it is not clear whether the drug reaches the proposed sites of action in vivo after oral ingestion, especially in humans. In this study, we examined the absorption, bioavailability, and metabolism of 14C-resveratrol after oral and i.v. doses in six human volunteers. The absorption of a dietary relevant 25-mg oral dose was at least 70%, with peak plasma levels of resveratrol and metabolites of 491 +/- 90 ng/ml (about 2 microM) and a plasma half-life of 9.2 +/ 0.6 h. However, only trace amounts of unchanged resveratrol (<5 ng/ml) could be detected in plasma. Most of the oral dose was recovered in urine, and liquid chromatography/mass spectrometry analysis identified three metabolic pathways, i.e., sulfate and glucuronic acid conjugation of the phenolic groups and, interestingly, hydrogenation of the aliphatic double bond, the latter likely produced by the intestinal microflora. Extremely rapid sulfate conjugation by the intestine/liver appears to be the rate-limiting step in resveratrol's bioavailability. Although the systemic bioavailability of resveratrol is very low, accumulation of resveratrol in epithelial cells along the aerodigestive tract and potentially active resveratrol metabolites may still produce cancer preventive and other effects. PMID- 15333515 TI - The chimpanzee (Pan troglodytes) as a pharmacokinetic model for selection of drug candidates: model characterization and application. AB - The chimpanzee (CHP) was evaluated as a pharmacokinetic model for humans (HUMs) using propranolol, verapamil, theophylline, and 12 proprietary compounds. Species differences were observed in the systemic clearance of theophylline (approximately 5-fold higher in CHPs), a low clearance compound, and the bioavailability of propranolol and verapamil (lower in CHPs), both high clearance compounds. The systemic clearance of propranolol (approximately 1.53 l/h/kg) suggested that the hepatic blood flow in CHPs is comparable to that in humans. No substantial differences were observed in the in vitro protein binding. A preliminary attempt was made to characterize cytochrome P450 (P450) activities in CHP and HUM liver microsomes. Testosterone 6beta-hydroxylation and tolbutamide methylhydroxylation activities were comparable in CHP and HUM liver microsomes. In contrast, dextromethorphan O-demethylation and phenacetin O-deethylation activities were approximately 10-fold higher (per mg protein) in CHP liver microsomes. Intrinsic clearance estimates in CHP liver microsomes were higher for propranolol (approximately 10-fold) and theophylline (approximately 5-fold) and similar for verapamil. Of the 12 proprietary compounds, 3 had oral clearances that differed in the two species by more than 3-fold, an acceptable range for biological variability. Most of the observed differences are consistent with species differences in P450 enzyme activity. Oral clearances of proprietary compounds in HUMs were significantly correlated to those from CHPs (r = 0.68; p = 0.015), but not to estimates from rat, dog, and monkey. In summary, the chimpanzee serves as a valuable surrogate model for human pharmacokinetics, especially when species differences in P450 enzyme activity are considered. PMID- 15333516 TI - Identification of critical amino acid residues of human CYP2A13 for the metabolic activation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, a tobacco-specific carcinogen. AB - Among all the known human cytochrome P450 enzymes, CYP2A13 has the highest efficiency in catalyzing the metabolic activation (keto aldehyde and keto alcohol formation) of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK), a potent lung carcinogen in animals and a suspected human lung carcinogen. As part of the structure-activity relationship (SAR) study, the present work was done to identify the key amino acid residues in CYP2A13 that are responsible for this high catalytic efficiency by using a series of mutants (Ala117Val, His164Gly, Ser208Ile, His372Arg, and Pro465Ser). In these CYP2A13 mutants, the amino acid residues were substituted by the residues at the corresponding positions of CYP2A6, which shares 93.5% amino acid sequence identity with CYP2A13 but is significantly less active (<5%) than CYP2A13 in NNK alpha-hydroxylation. We demonstrated that, except for the His164Gly mutant, all the CYP2A13 mutant proteins showed a significant decrease in the catalytic efficiency (Vmax/Km) for NNK alpha-hydroxylation. The His372 to Arg substitution resulted in a 20-fold increase in the Km value and a 7-fold decrease in the Vmax value for keto aldehyde formation as well as a total loss of detectable keto alcohol formation. The Ala117 to Val substitution, however, only caused a selective decrease in the Vmax value for keto aldehyde formation. The role of these amino acid residues in CYP2A13-catalyzed reactions is clearly substrate dependent, since the same Ala117Val and His372Arg mutants showed a 9-fold increase in the catalytic efficiency for coumarin 7-hydroxylation. Together with the computational substrate docking, our study provides new SAR in formation of human CYP2A13. PMID- 15333517 TI - A dual-color fluorescence imaging-based system for the dissection of antiangiogenic and chemotherapeutic activity of molecules. AB - We have developed a simple yet sensitive dual color fluorescence-based technique for dissecting the tumor-neovascularization relationship and evaluated the susceptibility of each component to therapeutic interventions. Green fluorescent protein (GFP)-expressing melanoma cells were cocultured with endothelial cells on different three-dimensional (3-D) matrices and exposed to multiple growth factors and molecules with established anti-angiogenic or anticancer activities. Cells were fixed and stained with propidium iodide, imaged using a confocal microscope, and stereologically analyzed. Three-dimensionality of the system was tested by depth-coding and pseudocolor 3-D reconstruction in the z-axis. Selective ablation of the tumor cells was affected by the anthracycline antibiotic doxorubicin. Treatment with vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) promoted the neovascular responses on matrigel and collagen-1 matrices. VEGF-induced angiogenesis was inhibited after treatment with combretastatin and thalidomide. In contrast, HGF exerted a protective effect against these anti-angiogenics in a matrigel matrix. However, this effect was lost when the matrix was substituted with collagen, suggesting that the extracellular matrix impinges on cellular function, possibly through an Akt mediated mechanism. The VEGF-receptor antagonist PTK787 also selectively ablated the VEGF-induced angiogenic effect without inhibiting the HGF-induced response, demonstrating the sensitivity of the system to detect modulation of distinct signal cascades. The current model encompasses the possibility of studying tumor angiogenesis-matrix interaction on the same platform, expanding the rapid screening of novel molecules in a simulated clinicopathological setting. PMID- 15333519 TI - Restructuring our workforce. PMID- 15333520 TI - Defining the specialty. PMID- 15333521 TI - Patient satisfaction in emergency medicine. AB - A systematic review was undertaken to identify published evidence relating to patient satisfaction in emergency medicine. Reviewed papers were divided into those that identified the factors influencing overall satisfaction in emergency department patients, and those in which a specific intervention was evaluated. Patient age and race influenced satisfaction in some, but not all, studies. Triage category was strongly correlated with satisfaction, but this also relates to waiting time. The three most frequently identified service factors were: interpersonal skills/staff attitudes; provision of information/explanation; perceived waiting times. Seven controlled intervention studies were found. These suggested that increased information on ED arrival, and training courses designed to improve staff attitudes and communication, are capable of improving patient satisfaction. None of the intervention studies looked specifically at the effect of reducing the perceived waiting time. Key interventions to improve patient satisfaction will be those that develop the interpersonal and attitudinal skills of staff, increase the information provided, and reduce the perceived waiting time. Future research should use a mixture of quantitative and qualitative methods to evaluate specific interventions. PMID- 15333522 TI - Development of the specialty of emergency medicine in Israel: comparison with the UK and US models. AB - OBJECTIVES: To describe the development of emergency medicine (EM) in Israel and review the specific problems faced by the discipline and describe the solutions that were found. METHODS: A comprehensive literature search was conducted for data on development of EM in the UK and in North America, and the personal knowledge of two of the authors (PH and YW) was used in preparing the article. RESULTS: There are differences in development of EM between Israel and the UK/US models. In Israel the specialty developed within the context of established high quality clinical practice and consequently it met resistance from the system, which did not wish to invest in what it felt might be marginal improvements in patient care. The economics of Israeli medicine also dictated that EM be made into a super-specialty rather than a primary specialty. Certified specialists from family medicine, paediatrics, internal medicine, general surgery, anaesthesia, and orthopaedic surgery can access training positions in EM. Currently there are seven active EM programmes of 2.5 years duration and 16 residents. The curriculum is flexible and a national certification examination is being developed. CONCLUSIONS: Development of EM can and should take different paths according to the specific local needs and realities. There is no single ideal model suitable for all circumstances. The practice of clinical EM in Israel is comparable with that of any developed country and daily progress is being made in the academic areas of teaching and research. There are worldwide similarities in the process of developing EM as a distinct discipline. PMID- 15333523 TI - Making an IMPACT on emergency department flow: improving patient processing assisted by consultant at triage. AB - OBJECTIVES: To assess whether initial patient consult by senior clinicians reduces numbers of patients waiting to be seen as an indirect measure of waiting time throughout the emergency department (ED). METHODS: An emergency medicine consultant and a senior ED nurse (G or F grade), known as the IMPACT team, staffed the triage area for four periods of four hours per week, Monday to Friday between 9 am to 5 pm for three months between December 2001 and February 2002 when staffing levels permitted. Patients normally triaged by a nurse in this area instead had an early consultation with the IMPACT team. Data were collected prospectively on all patients seen by the IMPACT team. The number of patients waiting to be seen (for triage, in majors and in minors) was assessed every two hours during the IMPACT sessions and at corresponding times when no IMPACT team was operational. RESULTS: There was an overall reduction in the number of patients waiting to be seen in the department from 18.3 to 5.5 (p<0.0001) at formal two hourly assessments. The largest difference was seen in minors. Of the patients seen at triage by the IMPACT team, 48.9% were discharged home immediately after assessment and treatment. With the IMPACT team present, no patient waited more than four hours for initial clinical consult. CONCLUSIONS: By using a senior clinical team for initial patient consultation, the numbers of patients waiting fell dramatically throughout the ED. Many patients can be effectively treated and discharged after initial consult by the IMPACT team. PMID- 15333524 TI - Team triage improves emergency department efficiency. AB - OBJECTIVE: To see whether three hours of combined doctor and nurse triage would lead to earlier medical assessment and treatment and whether this benefit would carry on for the rest of the day when normal triage had resumed. METHOD: Eight days were randomly selected; four for team triage and four for the normal nurse led triage. Team triage was coordinated by a middle grade or consultant from 9 am to 12 noon. Times to triage, to see a doctor, radiology, admission, and discharge were recorded. No additional medical or nursing staff were used and staffing levels were similar each day. All patients including blue light emergencies and minor injuries were included. RESULTS: Median times were significantly reduced (p<0.05) during the intervention to triage (2 min v 7 min, p = 0. 029), to see a doctor (2 min v 32 min, p = 0.029), and to radiology (11.5 min v 44.5 min, p = 0.029). Waiting times at midday were longer for patients in the non-intervention group. More patients were seen and discharged within 20 minutes in the intervention group (18 of 95 (19%) v 2 of 69 (3%) p = 0.0043). No significant knock on effect was demonstrable for the remaining 21 hours after the intervention ceased. CONCLUSION: Three hours of combined doctor and nurse triage significantly reduces the time to medical assessment, radiology, and to discharge during the intervention period. Waiting times at midday were shorter in the triage group. There was no significant knock on effect the rest of the day. PMID- 15333525 TI - Can a senior house officer's time be used more effectively? AB - OBJECTIVES: To determine the amount of time senior house officers (SHO) spent performing tasks that could be delegated to a technician or administrative assistant and therefore to quantify the expected benefit that could be obtained by employing such physicians' assistants (PA). METHODS: SHOs working in the emergency department were observed for one week by pre-clinical students who had been trained to code and time each task performed by SHOs. Activity was grouped into four categories (clinical, technical, administrative, and other). Those activities in the technical and administrative categories were those we believed could be performed by a PA. RESULTS: The SHOs worked 430 hours in total, of which only 25 hours were not coded due to lack of an observer. Of the 405 hours observed 86.2% of time was accounted for by the various codes. The process of taking a history and examining patients accounted for an average of 22% of coded time. Writing the patient's notes accounted for an average of 20% of coded time. Discussion with relatives and patients accounted for 4.7% of coded time and performing procedures accounted for 5.2% of coded time. On average across all shifts, 15% of coded time was spent doing either technical or administrative tasks. CONCLUSION: In this department an average of 15% of coded SHOs working time was spent performing administrative and technical tasks, rising to 17% of coded time during a night shift. This is equivalent to an average time of 78 minutes per 10 hour shift/SHO. Most tasks included in these categories could be performed by PAs thus potentially decreasing patient waiting times, improving risk management, allowing doctors to spend more time with their patients, and possibly improving doctors' training. PMID- 15333526 TI - An investigation into the short term and medium term health impacts of personal incapacitant sprays. A follow up of patients reported to the National Poisons Information Service (London). AB - OBJECTIVES: The aim of this study is to describe the pattern of ill health after personal incapacitant spray (PIS) exposures reported to the National Poisons Information Service-London (NPIS-L) and the Chemical Incident Response Service and to evaluate the relation between sub-categories of PIS exposure and adverse health effects. METHODS: Case series study of patients reported to the NPIS-L, by attending medical personnel during the period 16 January to 31 September 1998. Data collected by questionnaire sent to these medical personnel. RESULTS: Several "adverse" symptoms, particularly dermatitis and blisters were reported for cases exposed to police PIS. These cases were more frequent than in those people exposed to non-police PIS. Adverse effects occurring more than six hours after exposure were also observed, which is in conflict with the recorded immediate, short lived, and self limiting symptoms that PIS are designed to cause. Most patients with persisting symptoms required further treatment. CONCLUSIONS: These findings suggest that the formulation of CS (o-chlorobenzylidine malononitrile) with MiBK (methyl iso-butyl ketone) used by the police is more harmful that has been previously assumed. If confirmed then the continued use of this formulation should be reviewed because of longer duration of adverse effects. Less concentrated formulations may reduce the severity or persistence of the adverse effects. PMID- 15333527 TI - Effects of gastric acid on euro coins: chemical reaction and radiographic appearance after ingestion by infants and children. AB - OBJECTIVES: This study investigated whether coins of the new European currency (euro) corrode when they are exposed to gastric acid, and whether this change can be detected radiographically. METHODS: The eight different denominations of coins were immersed for seven days in 0.15 N hydrochloride acid (HCl), which corresponds to the level of post-prandial gastric acid. A Swedish crown coin and three different Austrian schilling coins were used as controls. The coins were weighed and radiographed daily to evaluate visible corrosions and HCl was analysed daily for possible dissolved substances. RESULTS: All coins lost weight within 24 hours after exposure to HCl. The 1, 2, and 5 euro cent coins developed changes that were visible on radiographs. The weights of all coins decreased by 0.43% to 11.30% during one week. The dissolved substances measured in the HCl corresponded to the different metals and alloys of the coins, except for copper, which does not dissolve in HCl. The highest absolute weight loss was observed in the Swedish crown coin (0.67 g), and the highest relative weight loss in the 1 Austrian schilling coin (11.30%). The two coins that showed the highest absolute and relative weight losses were the 2 euro (0.54 g or 6.35%) and the 1 euro (0.48 g or 6.39%) coin. CONCLUSIONS: A higher rate of toxicity for the new European coins compared with coins of other currencies is not expected, unless a massive coin ingestion occurs. PMID- 15333528 TI - End tidal carbon dioxide as a predictor of the arterial PCO2 in the emergency department setting. AB - OBJECTIVES: Patients arriving in the emergency department (ED) need rapid and reliable evaluation of their respiratory status. Mainstream end tidal carbon dioxide (ETCO(2)) is one of the methods used for this purpose during general anaesthesia of intubated patients in the operating theatre. Sidestream ETCO(2) (SSETCO(2)) might be a non-invasive, rapid, and reliable predictor of arterial Pco(2) in non-intubated patients in respiratory distress. The aim of this study was to verify whether SSETCO(2) can accurately predict the arterial Pco(2) and to detect variables that may affect this correlation. METHODS: A prospective semi blind study. The participants were 73 patients (47 men, 26 women) referred to the ED for respiratory distress. Arterial blood gas pressures and SSETCO(2) measurements were performed and recorded for all patients. Other parameters recorded were: age; body temperature; respiratory rate; blood pressure; pulse rate; and medical diagnosis. RESULTS: A significant correlation was found between SSETCO(2) and arterial Pco(2) (r = 0.792). Compared with the correlation curve of the whole group, age under 50 years deflected the correlation curve to the left, while temperature above 37.6 degrees C deflected it to the right. The rest of the parameters had no clear influence on the SSETCO(2)/Pco(2) correlation curve. CONCLUSIONS: There is a good correlation between SSETCO(2) and arterial Pco(2) in the ED setting. Young age may increase the arterial Pco(2)/SSETCO(2) gradient while raised temperature may decrease this gradient. Further studies are needed to confirm these findings in the normal healthy population. PMID- 15333529 TI - Non-touch suturing technique fails to reduce glove puncture rates in an accident and emergency department. AB - OBJECTIVE: To assess the impact of introducing a safer non-touch suturing technique into an inner city emergency department. METHODS: The rate of glove perforation, measured by electrical conductance, was used as a marker. Gloves (Bodyguards) used in suturing were collected over a two month period. Two half day suture workshops were then conducted in the emergency department and gloves were collected for a further two months. All doctors (14) were included in the study regardless of grade. Gloves were tested for perforation by electrical conductance. RESULTS: There were 107 gloves and 19 perforations in the first group, 133 with 28 perforations in the second (p = 0.52), and one perforation in 100 control gloves. Most doctors reported inadvertent needlestick handling. CONCLUSION: Two half day suturing workshops are not enough to reduce glove perforations from suturing in the emergency department. Difficulty with compliance suggests that safe suturing practice must be taught before doctors develop bad habits that are difficult to change. PMID- 15333530 TI - Detection of non-accidental injuries presenting at emergency departments. AB - OBJECTIVES: To investigate whether cases of possible non-accidental injury as identified using five risk indicators give rise to any subjective concerns of child abuse. METHODS: Questionnaires were completed by the triage nurse and attending doctor for every child attending the general hospitals of the North Western Health Board, with an injury, during a six month period. The questionnaires included an assessment of subjective concerns about the injury occurrence and five risk indicators of child abuse. RESULTS: Children presenting with an injury who had two or more positive indicators failed to raise subjective concerns in the attending emergency department staff. CONCLUSIONS: The introduction of a policy of identifying positive indicators from the five risk indicators of child abuse needs additional computer support within emergency departments. PMID- 15333531 TI - Cardiopulmonary resuscitation on Flemish television: challenges to the television effects hypothesis. AB - BACKGROUND: People who watch a lot of medical fiction overestimate the success rate of cardiopulmonary resuscitation (CPR). It has been suggested that this is because CPR is usually shown to be successful on television. This study analysed a popular Flemish medical drama series. Previous research showed that heavy viewing of this series was related to overestimation of CPR success. METHOD: Content analysis of 70 episodes of "Spoed" in the period between 2001 and the first three months of 2003. Causes and treatment of cardiac arrest and outcome of CPR were recorded in the same way as previous studies. RESULTS: CPR was performed 31 times in the 70 episodes. Only 19% of the patients survived the resuscitation attempt. Most patients were middle aged or older. Causes of arrest were different from those in British or American television series. CONCLUSIONS: The low survival rate challenges the idea that heavy viewers adopt the overestimation shown by television. Psychological research shows that people ignore base rate information in the shape of statistics, in favour of vivid, dramatic examples. Showing some impressive examples of success might therefore be more important than the overall success rate. It is suggested that the message of television fiction is that doctors are not powerless and that treatment does not stop once the heart stops beating. This helps to create what has been called an "illusion of efficacy". PMID- 15333532 TI - Are health professionals getting caught in the crossfire? The personal implications of caring for trauma victims. AB - OBJECTIVES: To investigate the long term psychological sequelae of treating multiple victims of traumatic incidents, such as violent crime and motor vehicle accidents, and to assess staff exposure to violent patients in the emergency department. METHODS: A self administered questionnaire booklet was distributed to all full time and part time staff working within the Johannesburg Hospital Trauma Unit during September 2002. Participation was voluntary. The questionnaire was specifically designed for the study as no relevant, validated questionnaire was found to be suitable. Psychological assessment comprised two standardised measures, the impact of event scale-revised and the Maslach burnout inventory. RESULTS: Thirty eight staff members completed the questionnaire, a response rate of 90%. Over 40% of respondents had been physically assaulted while at work and over 90% had been verbally abused. Staff reported a significant level of post traumatic symptoms, evaluated by the impact of event scale-revised (median = 17.5, range = 0-88), as a result of critical incidents they had been involved in during the previous six months. At least half of the respondents also reported a "high" degree of professional burnout in the three sub-scales of the Maslach burnout tnventory-that is emotional exhaustion, depersonalisation, and personal accomplishment. CONCLUSIONS: Preventative measures, such as increased availability of formal psychological support, should be considered by all trauma units to protect the long term emotional wellbeing of their staff. PMID- 15333533 TI - Quality and effectiveness of an emergency department during weekends. AB - OBJECTIVE: To evaluate whether the quality and effectiveness of an emergency department (ED) are modified during weekends. METHODS: Quality and effectiveness markers were determined during 539 consecutive days, comparing them according to the day of the week. Quality markers were the daily percentage of patients who died in the internal medical unit (deaths, D), leave ED without being seen (flights, F); returned to the ED (revisits, R), and the percentage of registered complaints (C). Effectiveness markers were: the "number of patients waiting to be seen" (WP), the "waiting time to be seen" (WT), and the "length of visit" (LV). RESULTS: Quality and effectiveness of ED do not worsen during weekend days and some markers significantly improved during such days: C experienced a 26% decrease (p = 0.001), WT decreased 65% (p<0.001), WP 59% (p<0.001), and LV 24% (p<0.01). Assessing the relation between daily number of visits to ED and the quality and effectiveness markers, a significant and direct association was found of the number of visits with D, F, R, and WP. CONCLUSION: Some of the quality and effectiveness markers of the ED improved during weekend days compared with workdays. PMID- 15333534 TI - Total time in English accident and emergency departments is related to bed occupancy. PMID- 15333535 TI - Are we training junior doctors to respond to major incidents? A survey of doctors in the Wessex region. AB - OBJECTIVE: To assess the current status of awareness and training of junior medical staff in the Wessex region in the event of a "conventional" major incident. METHODS: A telephone questionnaire of specialist registrars (SpRs) (or equivalent, for example, staff grade) in six core specialties was performed in all the 11 acute hospitals in the Wessex region on the same evening. This group was selected to represent a sample of the most senior medical staff "on site" at each hospital. RESULTS: 56 of 64 (87.5%) SpRs participated. Nine of the 56 (16%) SpRs questioned had previously been involved in a major incident, and 18 (32%) had experienced some form of major incident training exercise. Subgroup analysis of the specialties showed that although there were no significant differences in numbers of training experiences between specialties, only one of nine (11%) orthopaedic SpRs had ever been involved in a training exercise. Twenty five of the 56 (45%) SpRs felt that they were confident of their role in the event of an incident. CONCLUSION: Most middle grade staff in Wessex were not confident of their role in the event of a major incident. Most SpRs questioned had never attended a major incident training exercise. PMID- 15333536 TI - Prospective evaluation of non-radiologist performed emergency abdominal ultrasound for haemoperitoneum. PMID- 15333537 TI - Securing central venous catheters: a comparison of sutures with staples. AB - OBJECTIVES: Central venous access permits rapid drug delivery to the central circulation during cardiopulmonary resuscitation. Central venous catheters must be secured in place to prevent accidental removal and sutures are often used for this purpose. Staples may offer advantages over sutures by reducing the time needed to secure the central venous catheter and reducing exposure to contaminated sharps. The purpose of this study was to assess a staple anchoring device (Arrow, Reading, USA) and compare it with sutures for securing central venous catheters. METHODS: Prospective, randomised trial of 20 patients requiring insertion of a central venous catheter during elective surgery. RESULTS: Mean (SD) times were significantly shorter in the staples group than in the sutures group (staples 10.1 (3.5) seconds; sutures 50.5 (9.6) seconds; p<0.0001). However, 3 of 10 central venous catheters in the staples group were accidentally pulled out within the first three days. CONCLUSIONS: The use of staples significantly reduces the time to secure a central venous catheter. However, staples failed to secure the central venous catheters adequately and therefore cannot be recommended instead of sutures. PMID- 15333538 TI - SOCRATES 1 (synopsis of Cochrane reviews applicable to emergency services). PMID- 15333539 TI - Best evidence topic report. Nebulised magnesium in asthma. AB - A short cut review was carried out to establish whether the addition of nebulised magnesium sulphate to beta agonist therapy improves outcome in acute asthma. Altogether 69 papers were found using the reported search, of which five presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 15333541 TI - Best evidence topic report. Role of flexion/extension radiography in neck injuries in adults. AB - A short cut review was carried out to establish whether flexion-extension radiography is indicated in the investigation of a neurologically intact adult patient with midline neck tenderness and normal 3-view cervical spinal radiographs. Altogether 101 papers were found using the reported search, of which five presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 15333542 TI - Best evidence topic report. Peripheral pulses to exclude thoracic aortic dissection. AB - A short cut review was carried out to establish whether the absence of a clinical pulse deficit can be used to exclude dissecting thoracic aneurysm in patients with chest pain. Altogether 89 papers were found using the reported search, of which one was a previous systematic literature review. A further two papers published since the review were also found. These three papers presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 15333543 TI - Best evidence topic report. Wound closure in animal bites. AB - A short cut review was carried out to establish whether primary closure of animal bites increases wound infection rates. Altogether 74 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of this best paper are tabulated. A clinical bottom line is stated. PMID- 15333544 TI - Best evidence topic report. Therapeutic hypothermia after out of hospital cardiac arrest. AB - A short cut review was carried out to establish whether therapeutic hypothermia improves outcome in comatose post cardiac arrest patients. Altogether 176 papers were found using the reported search, of which four presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 15333545 TI - Best evidence topic report. Gastric lavage in aspirin and non-steroidal anti inflammatory drug overdose. AB - A short cut review was carried out to establish whether gastric lavage was better than activated charcoal alone at reducing toxicity after aspirin or other non steroidal anti-inflammatory drug (NSAID) overdose. Altogether 72 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. A further relevant paper was found on scanning the references of papers identified. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 15333546 TI - 6 Assessment and management of paediatric primary survey negative patients. PMID- 15333547 TI - 7 Abdominal pain, abdominal pain in women, complications of pregnancy and labour. PMID- 15333548 TI - The emerging role of the emergency care practitioner. AB - OBJECTIVES: To examine the emerging role of the emergency care practitioner (ECP) with comparisons to paramedic practice. Key activities were identified of newly appointed ECPs using qualitative methodology and a qualitative and quantitative comparison of patient treatment was made. METHOD: A constructivist methodology taking account of stakeholder input and drawing upon the constant comparisons of different group's construction of reality. Four practitioners completed reflective patient case studies and adapted patient report forms, which were compared with a second case group of 11 paramedics. In addition individual and focus groups interviews were performed with key stakeholders. RESULTS: In the comparison between ECP and paramedic roles, 331 paramedic incidents were compared with 170 ECP reports. ECPs treated 28% of patients on scene compared with 18% by paramedics (p = 0.007). Fifty per cent of ECPs patients were conveyed compared with 64% of paramedics (p = 0.000). Analysis of the 269 reflective reports and 14 stakeholder interviews revealed four key themes. Firstly, ECPs had a beneficial impact on the deployment of resources, especially relating to non-conveyance. Secondly, their training and education improved their decision making repertoire and developed their confidence for a leadership role. Thirdly, inter-agency collaboration and cooperation was improved, and finally, care benefits were increased especially relating to immediacy of treatment and referral mechanisms. CONCLUSIONS: The results indicate that an investment in the ECP role could be beneficial, however, more work is required to evaluate the development of practice, the quality of care, and cost benefits. PMID- 15333549 TI - Surviving out of hospital cardiac arrest at home: a postcode lottery? AB - OBJECTIVE: To find out if the response time, distance travelled to scene, and geographical location affect survival from out of hospital cardiac arrest (OHCA). METHODS: Retrospective cohort study over 10 years, 1 December 1991 to 1 August 2001. Outcome and demographic data were obtained for 1956 OHCAs occurring at home, in the Lothians region, from the Heartstart Scotland database. Survival rates to hospital admission and discharge were examined by postcode district. RESULTS: Certain postcode areas were served by quicker response times and shorter journeys. The survival to admission rate was greater in those areas where the median response time was <10 minutes (13.5% versus 8.1%, p<0.05). There was greater survival to discharge in these areas but not significantly so (4.1% versus 3.2%, p = 0.42). Survival to admission was more likely in areas where the median distance travelled was 90% alpha-exon inclusion in glioblastoma cells had no discernable effect on cell growth in culture, but was associated with an increase in unstimulated caspase-3 and -7 activity. The ability to manipulate endogenously expressed mRNA variants allows exploration of their functional relevance under normal and diseased physiological states. PMID- 15333584 TI - A synonymous SNP of the corneodesmosin gene leads to increased mRNA stability and demonstrates association with psoriasis across diverse ethnic groups. AB - Psoriasis is a chronic skin disorder with multifactorial aetiology. Genome-wide scans have provided unambiguous evidence for a major disease susceptibility locus on chromosome 6p21 (PSORS1). A minimal PSORS1 interval has been defined which encompasses three genes (HLA-C, HCR and CDSN) carrying psoriasis-associated SNPs. On the basis of this genetic evidence, we have undertaken an assessment of CDSN allele functional impact. A comparison of CDSN intragenic haplotypes showed that SNPs exclusive to disease-associated chromosomes are located in regions implicated in the stabilization of RNA transcripts. As CDSN is over-expressed in psoriatic lesions, we hypothesised that disease-associated intragenic SNPs may alter the rate of its mRNA decay. Here, we demonstrate that mRNAs transcribed from a CDSN risk haplotype present a 2-fold increase in stability, compared with those transcribed from a neutral haplotype (t-test P=0.004). Site-directed mutagenesis revealed that a single synonymous SNP (CDSN*971T) accounts for the observed increase in RNA stability. CDSN*971T maps to a RNA stability motif and UV cross-linking analysis demonstrated that the SNP affects the transcript affinity for a 39 kDa RNA binding protein. Association analyses show that haplotypes bearing CDSN*971T confer psoriasis susceptibility in a wide range of ethnic groups. These results demonstrate the effect of synonymous variation upon allele specific gene expression, a finding of relevance to future studies of the pathogenesis of common and complex traits. PMID- 15333585 TI - Analysis of DNA ligase IV mutations found in LIG4 syndrome patients: the impact of two linked polymorphisms. AB - LIG4 syndrome patients have hypomorphic mutations in DNA ligase IV. Although four of the five identified patients display immunodeficiency and developmental delay, one patient was developmentally normal. The developmentally normal patient had the same homozygous mutation (R278H) in DNA ligase IV as one of the more severely affected patients, who additionally had two linked polymorphisms. Here, we examine the impact of the mutations and polymorphisms identified in the LIG4 syndrome patients. Examination of recombinant mutant proteins shows that the severity of the clinical features correlates with the level of residual ligase activity. The polymorphisms decrease the activity of DNA ligase IV by approximately 2-fold. When combined with the otherwise mild R278H mutation, the activity is reduced to a level similar to other LIG4 patients who display immunodeficiency and developmental delay. This demonstrates how coupling of a mutation and polymorphism can have a marked impact on protein function and provides an example where a polymorphism may have influenced clinical outcome. Analysis of additional mutational changes in LIG4 syndrome (R580X, R814X and G469E) have led to the identification of a nuclear localization signal in DNA ligase IV and sites impacting upon DNA ligase IV adenylation. PMID- 15333586 TI - Common DNase I polymorphism associated with autoantibody production among systemic lupus erythematosus patients. AB - DNase I could be the most important nuclease for the removal of DNA from nuclear antigens at sites of high cell turnover, and thus may also prevent systemic lupus erythematosus (SLE). Sixteen SNPs were identified by direct DNA sequencing, among which six were selected for genotyping in a larger investigation on the basis of linkage disequilibria among SNPs, their frequency, location and haplotype tagging status. Genetic associations of polymorphisms in DNase I with the risk of SLE and the production of common autoantibodies were examined in a Korean population (350 SLE patients and 330 controls). Although no significant associations with the risk of SLE were found, logistic regression analyses revealed that one non synonymous SNPs in exon 8, +2373A>G(Gln244Arg), was significantly associated with an increased risk of the production of anti-RNP and anti-dsDNA antibodies among SLE patients. The frequency of the homozygous minor allele (Arg/Arg) was much higher in patients who had the anti-RNP antibody (31.3%) than in patients who did not have this antibody (14.4%) (P=0.0006, OR=2.86). In addition, the A/T mutation in exon 2 of DNase reported in two Japanese SLE patients was not present in SLE patients (n=350) or controls (n=330) in our Korean population, which combined with the results of previous reports strongly suggests that the mutation is not present in three major ethnic groups: Caucasian, African and Asian. PMID- 15333587 TI - Disease associations and altered immune function in CD45 138G variant carriers. AB - The CD45 antigen is a haemopoietic cell specific tyrosine phosphatase essential for antigen receptor mediated signalling in lymphocytes. Expression of different patterns of alternatively spliced CD45 isoforms is associated with distinct functions. We recently identified a polymorphism in exon 6 (A138G) of the gene encoding CD45 (PTPRC) that results in altered CD45 splicing. The 138G allele is present at a high frequency among Japanese (23.7%), with 5.1% individuals homozygous for the G allele. In this study we show that the A138G polymorphism is the cause of altered CD45 isoform expression, promoting splicing towards low molecular weight CD45 isoforms. We further report that the frequency of A138G heterozygotes is significantly reduced in number in cohorts of patients with autoimmune Graves' disease or hepatitis B infection, whereas G138G homozygotes are absent from a cohort of Hashimoto's thyroiditis patients. We also show that 138G individuals exhibit altered cytokine production in vitro and an increased proportion of memory T cells. These data suggest that the 138G variant allele strongly influences these diseases by modulation of immune mechanisms and may have achieved its high frequency as a result of a natural selection probably related to pathogen resistance. PMID- 15333588 TI - A molecular pathogenesis for transcription factor associated poly-alanine tract expansions. AB - Poly-alanine (Ala) tract expansions in transcription factors have been shown to be associated with human birth defects such as malformations of the brain, the digits, and other structures. Expansions of a poly-Ala tract from 15 to 22 (+7) 29 (+14) Ala in Hoxd13, for example, result in the limb malformation synpolydactyly in humans and in mice [synpolydactyly homolog (spdh)]. Here, we show that an increase of the Ala repeat above a certain length (22 Ala) is associated with a shift in the localization of Hoxd13 from nuclear to cytoplasmic, where it forms large amorphous aggregates. We observed similar aggregates for expansion mutations in SOX3, RUNX2 and HOXA13, pointing to a common mechanism. Cytoplasmic aggregation of mutant Hoxd13 protein is influenced by the length of the repeat, the level of expression and the efficacy of degradation by the proteasome. Heat shock proteins Hsp70 and Hsp40 co-localize with the aggregates and activation of the chaperone system by geldanamycin leads to a reduction of aggregate formation. Furthermore, recombinant mutant Hoxd13 protein forms aggregates in vitro demonstrating spontaneous misfolding of the protein. We analyzed the mouse mutant spdh, which harbors a +7 Ala expansion in Hoxd13 similar to the human synpolydactyly mutations, as an in vivo model and were able to show a reduction of mutant Hoxd13 and, in contrast to wt Hoxd13, a primarily cytoplasmic localization of the protein. Our results provide evidence that poly-Ala repeat expansions in transcription factors result in misfolding, degradation and cytoplasmic aggregation of the mutant proteins. PMID- 15333589 TI - An inducible null mutant murine model of Nijmegen breakage syndrome proves the essential function of NBS1 in chromosomal stability and cell viability. AB - The human genetic disorder, Nijmegen breakage syndrome, is characterized by radiosensitivity, immunodeficiency, chromosomal instability and an increased risk for cancer of the lymphatic system. The NBS1 gene codes for a protein, nibrin, involved in the processing/repair of DNA double strand breaks and in cell cycle checkpoints. Most patients are homozygous for a founder mutation, a 5 bp deletion, which might not be a null mutation, as functionally relevant truncated nibrin proteins are observed, at least in vitro. In agreement with this hypothesis, null mutation of the homologous gene, Nbn, is lethal in mice. Here, we have used Cre recombinase/loxP technology to generate an inducible Nbn null mutation allowing the examination of DNA-repair and cell cycle-checkpoints in the complete absence of nibrin. Induction of Nbn null mutation leads to the loss of the G2/M checkpoint, increased chromosome damage, radiomimetic-sensitivity and cell death. In vivo, this particularly affects the lymphatic tissues, bone marrow, thymus and spleen, whereas liver, kidney and muscle are hardly affected. In vitro, null mutant murine fibroblasts can be rescued from cell death by transfer of human nibrin cDNA and, more significantly, by a cDNA carrying the 5 bp deletion. This demonstrates, for the first time, that the common human mutation is hypomorphic and that the expression of a truncated protein is sufficient to restore nibrin's vital cellular functions. PMID- 15333590 TI - Syncytin: the major regulator of trophoblast fusion? Recent developments and hypotheses on its action. AB - Syncytin is a membrane protein derived from the envelope gene of an endogenous retrovirus of the HERV-W family. The gene appears to be almost exclusively expressed in placenta; the protein was found in particular in syncytiotrophoblast. After transfection into various cell types it has proven to be a very fusogenic protein, inducing the formation of syncytia. Therefore, the question rises as to whether syncytin is responsible for the fusion process of villous cytotrophoblast into syncytiotrophoblast in vivo. If so, how is this fusion process regulated if syncytin is found all over the syncytiotrophoblast? Can this process be regulated through local or temporal changes in syncytin expression, or is syncytin merely one factor in a cascade of events leading to fusion limited at some other level? This review will try to summarize the published data on the regulation of fusion in trophoblast models as well as on the localization and regulation of syncytin expression and of its presumed receptors. Assuming that syncytin is the key factor inducing trophoblast fusion, a number of models will be presented by which syncytin and/or its receptors might regulate this process. In some of the hypotheses proposed, local coexpression of syncytin and receptor, leading to blocking of one factor by the other, is of functional relevance. PMID- 15333591 TI - European Classification of Infertility Taskforce (ECIT) response to Habbema et al., 'Towards less confusing terminology in reproductive medicine: a proposal'. AB - The European Classification of Infertility Taskforce (ECIT) has been formed by ESHRE to develop a set of infertility-related codes (descriptions, interventions and outcomes) appropriate for computerization (http://www.ecit.info). In this article, ECIT reflects on previous suggestions in this Debate series, agreeing with the weaknesses of the current terminology used in reproductive medicine. Although ECIT is enthusiastic about the extension of terminology, caution is raised in abandoning familiar terms that may lead to greater rather than less confusion. Instead, ECIT suggests a way forward using structured computerized statements to retain current terms but extend their meaning. PMID- 15333592 TI - Effect of temperature upon adhesion formation in a laparoscopic mouse model. AB - BACKGROUND: Pneumoperitoneum can be a cofactor in adhesion formation. Pneumoperitoneum with non-humidified gas causes desiccation in the peritoneal cavity which decreases temperature. The effect of desiccation upon adhesion formation is widely accepted. The specific effect of the associated cooling upon adhesion formation remains unexplored, and was addressed specifically in our laparoscopic mouse model. METHODS: Adhesions were induced during laparoscopy and scored after 7 days during laparotomy. Pneumoperitoneum was performed using CO2 or CO2 with oxygen with or without humidification. Animals were placed at different environmental temperatures to modulate body and intraperitoneal temperature. RESULTS: Anaesthesia, environment with a lower temperature and pneumoperitoneum all independently decrease body temperature. A decrease in body temperature decreases adhesion formation (P=0.004). Therefore, at 37 degrees C, pneumoperitoneum-enhanced adhesion formation is more important than at room temperature (P=0.04). As was observed at room temperature, adhesion formation at 37 degrees C increases with the duration (P=0.01) of pneumoperitoneum and decreases with the addition of 3% of oxygen (P=0.03). CONCLUSIONS: Hypothermia reduces pneumoperitoneum-enhanced adhesion formation, which supports hypoxia as a driving mechanism, since hypothermia decreases the toxic effects of hypoxia and of the ischaemia-reperfusion process. These data could open up new possibilities for adhesion prevention in laparoscopic surgery. PMID- 15333593 TI - Ovarian function with the contraceptive vaginal ring or an oral contraceptive: a randomized study. AB - BACKGROUND: The effects on ovarian function of the combined contraceptive vaginal ring NuvaRing and a combined oral contraceptive (COC) were compared. METHODS: This randomized, open-label study was performed in 40 healthy female volunteers, who were randomized by a computer-generated list after stratification for the ovulation day in a pretreatment cycle. They received two cycles of NuvaRing (21 subjects) or a COC (30 microg ethinylestradiol and 150 microg levonorgestrel, 19 subjects). NuvaRing was started on cycle day 5, COC on cycle day 1. Follicular diameter, endometrial thickness and FSH, LH, 17beta-estradiol (E2) and progesterone concentrations were determined. RESULTS: The median maximum follicular diameter (maxFD) was < or =11 mm during treatment. In the first treatment cycle the maxFD was lower in the COC than in the NuvaRing group, due to the different starting procedures. MaxFD were not different in the second treatment cycle. In both groups, E2 and progesterone levels remained low during treatment. Ovulations did not occur. CONCLUSIONS: In both groups, ovarian activity was adequately suppressed. Due to the different starting procedures, lower ovarian activity was observed in the COC group in the first treatment cycle. In the second cycle, ovarian suppression was comparable with NuvaRing and COC treatment. PMID- 15333594 TI - From spermatocytes to sperm: meiotic behaviour of human male reciprocal translocations. AB - BACKGROUND: Human male translocation carriers may present alterations in the meiotic process due to the presence of the translocated chromosomes. The aim of this work was to study the mechanisms that affect meiotic segregation in translocation carriers by analysing different stages of the meiotic process. METHODS: Meiotic studies using fluorescence in-situ hybridization on both spermatocytes and sperm nuclei were performed in two translocation carriers, t(11;17)(q13.1;p11.2) and t(10;14)(q24;q32). RESULTS: A ring configuration was the main type of quadrivalent found in metaphase I. Overall chiasma frequency was significantly decreased in the t(11;17) carrier. In the t(10;14) carrier, chiasma frequency within the interstitial region of chromosomes 10 and 14 was increased and the recombination pattern was modified. As expected from the frequencies of interstitial chiasmata found in metaphase I in the two subjects, the incidence of asymmetric dyads was sporadic in t(11;17) and very high in t(10;14). In both carriers, segregation frequencies observed at metaphase II were not different from the segregation data obtained in decondensed sperm nuclei. CONCLUSIONS: The concordance observed among results obtained in different spermatogenic stages indicates an absence of cellular selection based on chromosomal imbalances. Results obtained in the aneuploidy assay have not provided any evidence for an interchromosomal effect. PMID- 15333595 TI - Depot testosterone with etonogestrel implants result in induction of azoospermia in all men for long-term contraception. AB - BACKGROUND: Combined testosterone and progestogen preparations are a promising approach to male hormonal contraception. We investigated the effect of s.c. etonogestrel with depot testosterone on spermatogenesis in normal men over a period of 48 weeks. METHODS: Fifteen healthy men received three s.c. 68 mg etonogestrel implants. Testosterone pellets (400 mg) were administered at 12 weekly intervals. RESULTS: Nine men completed 48 weeks of treatment. Four subjects chose to discontinue after 6 months, one man withdrew from the study early for personal reasons and one was withdrawn due to illness. Sperm concentrations of <1 x 10(6)/ml were achieved in all men by 16 weeks of treatment. All men became azoospermic, although the time to achieve this varied from 8 to 28 weeks. Azoospermia was maintained in eight of the nine men treated for 48 weeks, one subject showing partial recovery from 40 weeks. Testosterone levels remained in the physiological range throughout. Treatment did not result in weight gain, change in body composition or decline in high-density lipoprotein cholesterol concentrations. CONCLUSIONS: The combination of three etonogestrel implants with depot testosterone results in rapid and consistent suppression of spermatogenesis. This can be maintained for up to 1 year and may therefore be a suitable approach for a long-acting male hormonal contraceptive. PMID- 15333596 TI - Maternal KIR repertoire is not associated with recurrent spontaneous abortion. AB - BACKGROUND: In view of evidence suggesting an immunological cause of recurrent spontaneous abortions (RSA) and the large number of maternal natural killer (NK) cells present in the pregnant uterus, we investigated the genetic polymorphism of the killer cell immunoglobulin-like receptors (KIR) in women with RSA. METHODS: KIR gene repertoire and KIR2DL4 (a receptor for HLA-G) genotyping were determined by SSP and SSCP respectively, in women experiencing RSA and controls. RESULTS: The KIR repertoire did not differ between RSA patients and controls in terms of: (i) the number of inhibitory receptors; (ii) the number of activating receptors; (iii) the ratio of inhibitory to activating receptors. KIR2DL4, a receptor for HLA-G, has different transmembrane alleles, which produce functionally different phenotypes. The frequency of KIR2DL4 transmembrane genotypes differed significantly between RSA patients and controls (P=0.03). However, although homozygosity for a membrane-bound receptor was more frequent in patients (25%) than controls (10%), other genotypes that would produce the same phenotype were not more frequent in patients than controls. CONCLUSIONS: The data provide little evidence that KIR polymorphism plays a role in predisposition to RSA. PMID- 15333597 TI - Association of CYP1A1 gene polymorphism with recurrent pregnancy loss in the South Indian population. AB - BACKGROUND: We investigated the relationship between idiopathic recurrent pregnancy loss (RPL) and genetic polymorphisms in phase I and phase II detoxification genes which include CYP1A1, CYP2D6, GSTM1, GSTP1 and GSTT1. METHOD: A case-control study comprised 160 females with RPL and 63 healthy controls with a successful reproductive history. RESULTS: The CYP1A1 variant allele was present at frequencies of 0.61 and 0.44 in cases and controls, respectively (odds ratio=1.93; P=0.023, 95% confidence interval 1.10-3.38). The CYP2D6 variant allele was present at a frequency of 0.17 in females with RPL, while in the control population the frequency was 0.16. The GSTM1 and GSTT1 null genotypes were present at frequencies of 0.39 and 0.26 in RPL cases, whereas in controls the frequencies were 0.37 and 0.17, respectively. The mutant GSTP1 frequencies in case and control women were 0.38 and 0.40, respectively. We report a significant association of the CYP1A1*2A allele with RPL which is confirmed by logistic regression analysis. No association was observed for the other polymorphisms or in their combinations studied. CONCLUSIONS: The present study suggests the occurrence of the CYP1A1*2A allele as a probable risk factor in idiopathic recurrent miscarriages. PMID- 15333598 TI - Characterization of cystic fibrosis conductance transmembrane regulator gene mutations and IVS8 poly(T) variants in Portuguese patients with congenital absence of the vas deferens. AB - BACKGROUND: Cystic fibrosis conductance transmembrane regulator (CFTR) gene mutations and IVS8 poly(T) variants in Portuguese patients with bilateral (CBAVD) and unilateral (CUAVD) congenital absence of the vas deferens remain to be evaluated. METHODS: Patient screening was carried out by PCR, denaturing gradient gel electrophoresis and DNA sequencing. RESULTS: CFTR mutations were found in 18 out of 31 (58.1%) CBAVD and in three of four (75%) CUAVD patients. The most frequent mutations were F508del and R334W in CBAVD and G542X in CUAVD, with the allelic frequencies of R334W (6.5%) and G542X (25%) being particular to the Portuguese population. The 5T allelic frequency was 3.5% in the fertile male population, 25% in CUAVD and 27.4% in CBAVD patients. The combined frequency of mutations (CFTR+5T) was increased in CBAVD to 22 out of 31 (71%). The frequency of CFTR mutations was compared with that of patients with secondary obstructive azoospermia (OAZ; one out of 16, 6.3%) and non-obstructive azoospermia (NOAZ; two out of 22, 9.1%) with conserved spermatogenesis, which were similar to the general population. However, whereas the 5T allelic frequency in OAZ was similar to that of the general population (3.1%), it was increased in NOAZ cases (14.3%). CONCLUSIONS: Data confirm that CFTR+5T mutations represent the most common genetic abnormality in CAVD, and suggest that cases of NOAZ may be associated with the 5T allele. PMID- 15333599 TI - A synthetic analogue of meiosis-activating sterol (FF-MAS) is a potent agonist promoting meiotic maturation and preimplantation development of mouse oocytes maturing in vitro. AB - BACKGROUND: Follicular fluid-meiosis-activating sterol (FF-MAS) is a factor present in the pre-ovulatory follicle during the time of oocyte maturation. In mouse oocytes maturing in vitro, FF-MAS promotes the completion of meiotic maturation to metaphase II (MII) and improves competence to complete the 2-cell stage to blastocyst transition. We produced analogues of FF-MAS and selected three on the basis of potency to promote the resumption of meiosis by mouse oocytes maintained in meiotic arrest by hypoxanthine. The objective of this study was to determine whether these FF-MAS analogues also affect the quality of oocytes maturing in vitro with respect to the completion of meiotic maturation and augmenting the frequency of development to the blastocyst stage after fertilization in vitro. METHODS: Cumulus cell-enclosed oocytes were isolated from the small antral follicles of 18 or 20 day post-natal mice. These oocytes normally have a reduced competence to complete meiotic maturation and preimplantation embryo development. Oocytes were isolated at the germinal vesicle stage and matured in vitro using media supplemented with 0.1% ethanol, 1 micromol/l FF-MAS, or 0.1-10 micromol/l FF-MAS analogues ZK255884 (884), ZK255933 (933) and ZK255991 (991). Oocytes that progressed to MII were fertilized in vitro and the percentage developing to the 2-cell and blastocyst stages was determined. RESULTS: At 1 micromol/l, 991 and 933 increased the portion of oocytes progressing to MII, whereas the lowest dose of 991 and 884 was ineffective. Treatment of maturing oocytes with either 0.1 or 1 micromol/l 933 dramatically increased oocyte competence to complete preimplantation development. CONCLUSIONS: The synthetic analogue of FF-MAS, ZK255933, is a potent agonist that improves the quality of mouse oocytes matured in vitro. This compound may therefore have therapeutic value for treatment of oocytes from women undergoing therapy for infertility owing to poor oocyte quality. PMID- 15333600 TI - Does ovarian hyperstimulation in intrauterine insemination for cervical factor subfertility improve pregnancy rates? AB - BACKGROUND: Intrauterine insemination (IUI) can be performed with or without controlled ovarian hyperstimulation (COH). Studies in which the additional benefit of COH on IUI for cervical factor subfertility is assessed are lacking. We assessed whether COH in IUI improved pregnancy rates in cervical factor subfertility. METHODS: We performed a historical cohort study among couples with cervical factor subfertility, treated with IUI. A cervical factor was diagnosed by a well-timed, non-progressive post-coital test with normal semen parameters. We compared ongoing pregnancy rate per cycle in groups treated with IUI with or without COH. We tabulated ongoing pregnancy rates per cycle number and compared the effectiveness of COH by stratified univariable analysis. RESULTS: We included 181 couples who underwent 330 cycles without COH and 417 cycles with COH. Ongoing pregnancy rates in IUI cycles without and with COH were 9.7% and 12.7%, respectively (odds ratio 1.4; 95% confidence interval 0.85-2.2). The pregnancy rates in IUI without COH in cycles 1, 2, 3 and 4 were 14%, 11%, 6% and 15%, respectively. For IUI with COH, these rates were 17%, 15%, 14% and 16%, respectively. CONCLUSIONS: Although our data indicate that COH improves the pregnancy rate over IUI without COH, IUI without COH generates acceptable pregnancy rates in couples with cervical factor subfertility. Since IUI without COH bears no increased risk for multiple pregnancy, this treatment should be seriously considered in couples with cervical factor subfertility. PMID- 15333601 TI - Infertility caused by PCOS--health-related quality of life among Austrian and Moslem immigrant women in Austria. AB - BACKGROUND: The polycystic ovarian syndrome (PCOS) is the most common endocrine disorder affecting female fertility. In this study, we examined the symptomatology of PCOS and the health-related quality of life among infertility patients suffering from PCOS with different socio-cultural and ethnic background. METHODS: Symptomatology of PCOS, body composition characteristics as well as socio-economic factors were examined in 49 PCOS infertility patients of the University Clinic for Gynecology and Obstetrics, in Vienna, who originated from two socio-cultural subgroups, Austrian women and Moslem immigrant women. Additionally, the Cronins health-related quality of life questionnaire of women with PCOS was used in order to examine the impact of PCOS symptoms on the individual quality of life of the affected women. RESULTS: In terms of the appearance of the symptoms, the typical heterogeneity of PCOS could be found in both subgroups with no differences. However, differences in the health-related quality of life were impressive. Health-related quality of life of women from an Islamic background was affected to a greater degree than that of Austrian women, although no differences in symptomatology were found. This was true of all five domains investigated (infertility, overweight, hirsutism, menstrual irregularities and emotional problems). First of all, infertility was a dramatic problem for immigrant women. Islamic women had very high reproductive pressure. The Moslem immigrant PCOS women suffer more from infertility than do Austrian women. CONCLUSIONS: Health professionals should be sensitive to the ethnicity, religious and cultural background of their patients to provide the best possible medical support. PMID- 15333602 TI - Effect of advanced provision of emergency contraception on women's contraceptive behaviour: a randomized controlled trial. AB - BACKGROUND: Emergency contraception (EC) can prevent pregnancy but is under-used. Advanced provision increases use but the effect on contraceptive behaviour varies. METHODS: Women aged 18-45 years, using less effective contraceptives, were randomized to either advanced provision of three courses of EC (intervention) or to obtaining each course from clinic (control). EC use and contraceptive behaviour were monitored for 1 year. RESULTS: In all, 1030 women were recruited in 6 months. The mean+/-SD number of courses of EC used in intervention versus control group was 0.56+/-1.2 versus 0.20+/-0.6 (P<0.001). In the intervention group, 47% women aged <26 years used at least one course of EC compared with 23% of older women (P<0.001). The majority of women used condoms before (intervention 89%, control 91%) and during the study (89% for both groups). Consistency of contraceptive use was higher during the study (65 versus 60% of women in both groups) (P<0.001). There were 17 unplanned pregnancies, eight in the intervention group, six of whom did not use EC in the conception cycle. CONCLUSIONS: Advanced provision increases EC use especially among young women in Hong Kong. Contraceptive choice and consistency of use remains the same even among young women. PMID- 15333603 TI - Roles of p38 and c-jun in the differentiation, proliferation and immortalization of normal human endometrial cells. AB - BACKGROUND: It has been reported that p38 and c-jun operate as mediators of cell proliferation and differentiation. Therefore, by studying the roles of c-jun and p38 in the proliferation and differentiation of normal human endometrial cells, we can better understand the mechanism of these processes in endometrial cells. METHODS: Separation of glandular and stromal components was based on a modification of the work of Satyaswaroop et al. To confirm the purification of the endometrial cells and the expression of the transfected SV40 large T antigen, immunocytochemical analysis and western blot analysis were performed. RESULTS: There were polygonal shapes in the stromal cells in the early passage 1-2, while the aged endometrial stromal cells were spindle shaped. To investigate passage dependent molecular events in endometrial cells, the c-jun and pp38 levels were examined. Both c-jun and pp38 were significantly reduced with cellular aging and passages. To understand the role of c-jun, endometrial stromal cells were treated with SP600125 which is a specific inhibitor of c-jun. SP600125 induced morphological changes of young endometrial stromal cells with polygonal shape; the young cells appeared as aged endometrial cells with spindle shape. In addition, an immortalized endometrial cell line was established and shown to express activated c-jun, similiar to normal endometrial cells. CONCLUSIONS: These results suggest that the modulation of p38 and c-jun may play an important role in the differentiation and proliferation of human endometrial cells. PMID- 15333604 TI - Complex malformations of the female genital tract. New types and revision of classification. AB - BACKGROUND: Complex malformations of the female genital tract are often incorrectly identified, treated and reported, probably due to not considering the malformation as a cause of the clinical symptoms and neither the embryological origin of the different elements of the genitourinary tract. METHODS: Complex malformations are studied and classified, and new types are presented. The new types of complex malformations presented are: (i) Cases of unilateral vaginal or cervico-vaginal atresia with renal agenesis and uterine duplication, with or without communication between hemiuteri; (ii) the unilateral Rokitansky syndrome; and (iii) the combination in the same patient of unilateral Rokitansky syndrome (Mullerian defect) on one side and blind vagina and ipsilateral renal agenesis syndrome (Wolffian defect) on the other side. RESULTS: A revised version of the clinical and embryological classification of genital malformations is presented and an associated diagram points out the origin of these malformations. CONCLUSIONS: These genital malformative anomalies reaffirm our hypothesis about the embryology of the human vagina as deriving from the Wolffian ducts and the Mullerian tubercle; and they show that gynecologists should be aware of the related symptoms and the embryology of the female genital tract in order to achieve a better comprehension of the malformations for their right correction or therapeutic approach. PMID- 15333605 TI - Dose-finding study of daily gonadotropin-releasing hormone (GnRH) antagonist for the prevention of premature luteinizing hormone surges in IVF/ICSI patients: antide and hormone levels. AB - BACKGROUND: The aim of this study was to define the minimal effective dose of antide (Iturelix) to prevent premature luteinizing hormone (LH) surges in in vitro fertilization (IVF) patients. METHODS: In a prospective, single centre study, 144 IVF/ICSI patients were stimulated with r-hFSH from cycle day 2 and from cycle day 6 onwards, cotreated with daily 2 mg/2 ml (n=30), 1 mg/ml (n=30), 0.5 mg/ml (n=31), 0.5 mg/0.5 ml (n=23) and 0.25 mg/ml (n=30) GnRH antagonist (antide). Serum samples were taken three times daily during antide administration to assess antide and hormone levels. The minimal effective dose was defined as the lowest dose group with <2 LH surges (LH >12.4 IU/l and progesterone >2 ng/ml). RESULTS: Serum antide levels, mean LH and E2 levels per day and their area under the curves were dose-related to antide. The bioavailability of antide almost doubled after dilution in larger volumes. Pre-injection LH levels gradually increased during GnRH antagonist treatment. LH surges occurred in the lowest dose groups 0.5 mg/ml (3.2%), 0.5 mg/0.5 ml (6.7%) and 0.25 mg/ml (13.3%). Hence, 0.5 mg/ml is considered to be the minimal effective dose. Antide was overall well tolerated and safe. CONCLUSIONS: 0.5 mg/ml antide is the minimal effective dose to prevent an untimely LH surge in IVF patients stimulated with r hFSH. PMID- 15333606 TI - Urinary levels of insecticide metabolites and DNA damage in human sperm. AB - BACKGROUND: Members of the general population are exposed to non-persistent insecticides at low levels. The present study explored whether environmental exposures to carbaryl and chlorpyrifos are associated with DNA damage in human sperm. METHODS: Subjects (n=260) were recruited through a Massachusetts infertility clinic. Individual exposures were measured as spot urinary metabolite concentrations of chlorpyrifos [3,5,6-trichloro-2-pyridinol (TCPY)] and carbaryl [1-naphthol (1N)], adjusted using specific gravity. Sperm DNA integrity was assessed by neutral comet assay and reported as comet extent, percentage DNA in comet tail (Tail%) and tail distributed moment (TDM). RESULTS: A statistically significant increase in Tail% was found for an interquartile range (IQR) increase in both 1N [coefficient=4.1; 95% confidence interval (CI) 1.9-6.3] and TCPY (2.8; 0.9-4.6), while a decrease in TDM was associated with IQR changes in 1N (-2.2; 4.9 to 0.5) and TCPY (-2.5; -4.7 to -0.2). A negative correlation between Tail% and TDM was present only when stratified by comet extent, suggesting that Tail% and TDM may measure different types of DNA damage within comet extent strata. CONCLUSIONS: Environmental exposure to carbaryl and chlorpyrifos may be associated with increased DNA damage in human sperm, as indicated by a change in comet assay parameters. PMID- 15333607 TI - Cervical priming prior to operative hysteroscopy: a randomized comparison of laminaria versus misoprostol. AB - BACKGROUND: We aimed to compare efficacy of intravaginal misoprostol versus endocervical laminaria tents prior to operative hysteroscopy in selected cases. METHODS: A total of 144 patients with diagnosed intrauterine lesions scheduled for operative hysteroscopy were randomly allocated to two groups according to method of cervical priming prior to the procedure. Misoprostol 200 microg was inserted into the posterior fornix of the vagina for patients in group A (n=72), while laminaria tents were inserted intracervically in group B patients (n=72). RESULTS: Both methods were effective for cervical dilatation with a mean cervical diameter of 7.5+/-1.2 and 7.6+/-1.2 mm respectively. There was no significant difference in the mean cervical diameter or the time required for cervical dilatation (51.6 versus 51.4 s respectively). In contrast, there was a significant difference between the groups with respect to the insertion difficulty and in doctors' and patients' assessments of the procedure. CONCLUSIONS: Both misoprostol and laminaria were equally effective in inducing proper cervical priming prior to operative hysteroscopy with minimal time of cervical dilatation. Nevertheless, misoprostol may be superior due to easy application, reduced cost, and patient convenience and acceptability. PMID- 15333608 TI - Strenuous physical exercise inhibits granulocyte activation induced by high altitude. AB - To test the hypothesis of whether strenuous physical exercise inhibits neutrophils that can get activated by hypobaric hypoxia, we analyzed the effects of both high altitude and strenuous exercise alone and in combination on potentially cytotoxic functions of granulocytes in healthy volunteers (n = 12 men; average age 27.6 yr; range 24-38 yr). To this end, a field study was prospectively performed with an open-labeled within-subject design comprising three protocols. Protocol I (high altitude) involved a helicopter ascent, overnight stay at 3,196 m, and descent on the following day. Protocol II (physical exercise) involved hiking below an altitude of 2,100 m with repetitive ascents amounting to a total ascent to that of protocol III. Protocol III (combination of physical exercise and high altitude) involved climbing from 1,416 to 3,196 m, stay overnight, and descent on the following day. In protocol I, number of granulocytes did not change, but potentially cytotoxic functions of cells (CD18 expression and superoxide production) were early and significantly upregulated. In protocol II, subjects developed granulocytosis, but functions of cells were inhibited. In protocol III, granulocytosis occurred at higher values than those observed under protocol II. Potentially cytotoxic functions of cells, however, were strongly inhibited again. In conclusion, high altitude alone, even moderate in extent, can activate potentially cytotoxic functions of circulating granulocytes. Strenuous physical exercise strongly inhibits this activation, which may give protection from an otherwise inflammatory injury. PMID- 15333609 TI - Effects of acute creatine kinase inhibition on metabolism and tension development in isolated single myocytes. AB - This study investigated the effects of acute creatine kinase (CK) inhibition (CKi) on contractile performance, cytosolic Ca2+ concentration ([Ca2+]c), and intracellular PO2 (PIO2) in Xenopus laevis isolated myocytes during a 2-min bout of isometric tetanic contractions (0.33-Hz frequency). Peak tension was similar between trials during the first contraction but was significantly (P < 0.05) attenuated for all subsequent contractions in CKi vs. control (Con). The fall in PIO2 (DeltaPIO2) from resting values was significantly greater in Con (26.0 +/- 2.2 Torr) compared with CKi (17.8 +/- 1.8 Torr). However, the ratios of Con to CKi end-peak tension (1.53 +/- 0.11) and DeltaPO2 (1.49 +/- 0.11) were similar, suggesting an unaltered aerobic cost of contractions. Additionally, the mean response time (MRT) of DeltaPIO2was significantly faster in CKi vs. Con during both the onset (31.8 +/- 5.5 vs. 49.3 +/- 5.7 s; P < 0.05) and cessation (21.2 +/ 4.1 vs. 68.0 +/- 3.2 s; P < 0.001) of contractions. These data demonstrate that initial phosphocreatine hydrolysis in single skeletal muscle fibers is crucial for maintenance of sarcoplasmic reticulum Ca2+ release and peak tension during a bout of repetitive tetanic contractions. Furthermore, as PIO2 fell more rapidly at contraction onset in CKi compared with Con, these data suggest that CK activity temporally buffers the initial ATP-to-ADP concentration ratio at the transition to an augmented energetic demand, thereby slowing the initial mitochondrial activation by mitigating the energetic control signal (i.e., ADP concentration, phosphorylation potential, etc.) between sites of ATP supply and demand. PMID- 15333610 TI - Physiological basis of muscle functional MRI: predictions using a computer model. AB - Muscle functional MRI (mfMRI) has been proposed as a tool for noninvasively measuring the metabolic and hemodynamic responses to muscle activation, but its theoretical basis remains unclear. One challenge is that it is difficult to isolate individually those variables affecting the magnitude and temporal pattern of the mfMRI response. Therefore, the purpose of this study was to develop a computer model of how physiological factors altered during exercise affect the mfMRI signal intensity time course and then predict the contributions made by individual factors. A model muscle containing 39,204 fibers was defined. The fiber-type composition and neural activation strategies were designed to represent isometric contractions of the human anterior tibialis muscle, for which published mfMRI data exist. Sustained isometric contractions at 25 and 40% maximum voluntary contraction were modeled, as were the vascular (capillary recruitment, blood oxygen extraction) and metabolic (lactate accumulation, phosphocreatine hydrolysis, pH) responses. The effects on the transverse relaxation of MRI signal were estimated, and the mfMRI signal intensity time course was measured from simulated images. The model data agreed well qualitatively with published experimental data, and at long exercise durations the quantitative agreement was also good. The model was then used to predict that NMR relaxation effects secondary to blood volume and oxygenation changes, plus the creatine kinase reaction, dominate the mfMRI time course at short exercise durations (up to approximately 45 s) and that effects secondary to glycolysis are the main contributors at later times. PMID- 15333611 TI - Quantitative models of the rat pulmonary arterial tree morphometry applied to hypoxia-induced arterial remodeling. AB - Little is known about the constituent hemodynamic consequences of structural changes that occur in the pulmonary arteries during the onset and progression of pulmonary arterial remodeling. Many disease processes are known to be responsible for vascular remodeling that leads to pulmonary arterial hypertension, cor pulmonale, and death. Histology has been the primary tool for evaluating pulmonary remodeling, but it does not provide information on intact vascular structure or the vessel mechanical properties. This study is an extension of our previous work in which we developed an alternative imaging technique to evaluate pulmonary arterial structure. The lungs from Sprague-Dawley rats were removed, perfusion analysis was performed on the isolated lungs, and then an X-ray contrast agent was used to fill the arterial network for imaging. The lungs were scanned over a range of intravascular pressures by volumetric micro-computed tomography, and the arterial morphometry was mapped and measured in the reconstructed isotropic volumes. A quantitative assessment of hemodynamic, structural, and biomechanical differences between rats exposed for 21 days to hypoxia (10% O(2)) or normoxia (21.0% O(2)) was performed. One metric, the normalized distensibility of the arteries, is significantly (P < 0.001) larger [0.025 +/- 0.0011 (SE) mmHg(-1)] (n = 9) in normoxic rats compared with hypoxic [0.015 +/- 0.00077 (SE) mmHg(-1)] (n = 9). The results of the study show that these models can be applied to the Sprague-Dawley rat data and, specifically, can be used to differentiate between the hypoxic and the control groups. PMID- 15333612 TI - A high-fat, refined-carbohydrate diet induces endothelial dysfunction and oxidant/antioxidant imbalance and depresses NOS protein expression. AB - We tested whether consumption of a high-fat, high-sucrose (HFS) diet can affect endothelium-dependent relaxation, whether this precedes the development of diet induced hypertension previously noted in this model, and whether it is mediated, in part, by changes in nitric oxide synthase (NOS) and/or NOS regulatory proteins. Female Fischer rats were fed either a HFS diet or standard low-fat, complex-carbohydrate chow starting at 2 mo of age for 7 mo. Vasoconstrictive response to KCl and phenylephrine was similar in both groups. Vasorelaxation to acetylcholine was significantly impaired in the HFS animals, and there were no differences in relaxation to sodium nitroprusside, suggesting that the endothelial dysfunction is due, at least in part, to nitric oxide deficiency. HFS consumption decreased protein expression of endothelial NOS in aorta, renal, and heart tissues, neuronal NOS in kidney, heart, aorta, and brain, and inducible NOS in heart and aorta. Caveolin-1 and soluble guanylate cyclase protein expression did not change, but AKT protein expression decreased in heart and aorta and increased in kidney tissue. Consumption of HFS diet raised brain carbonyl content and plasma hydrogen peroxide concentration and diminished plasma total antioxidant capacity. Because blood pressure, which is known to eventually rise in this model, was not as yet significantly elevated, the present data suggest that endothelial dysfunction precedes the onset of diet-induced hypertension. The lack of a quantitative change in caveolin-1 and soluble guanylate cyclase protein content indicates that alteration in these proteins is not responsible for the endothelial dysfunction. Thus nitric oxide deficiency combined with antioxidant/oxidant imbalance, appears to be a primary factor in the development of endothelial dysfunction in this model. PMID- 15333613 TI - Active and passive components in the length-dependent stiffness of tracheal smooth muscle during isotonic shortening. AB - Contraction of smooth muscle tissue involves interactions between active and passive structures within the cells and in the extracellular matrix. This study focused on a defined mechanical behavior (shortening-dependent stiffness) of canine tracheal smooth muscle tissues to evaluate active and passive contributions to tissue behavior. Two approaches were used. In one, mechanical measurements were made over a range of temperatures to identify those functions whose temperature sensitivity (Q(10)) identified them as either active or passive. Isotonic shortening velocity and rate of isometric force development had high Q(10) values (2.54 and 2.13, respectively); isometric stiffness showed Q(10) values near unity. The shape of the curve relating stiffness to isotonic shortening lengths was unchanged by temperature. In the other approach, muscle contractility was reduced by applying a sudden shortening step during the rise of isometric tension. Control contractions began with the muscle at the stepped length so that properties were measured over comparable length ranges. Under isometric conditions, redeveloped isometric force was reduced, but the ratio between force and stiffness did not change. Under isotonic conditions beginning during force redevelopment at the stepped length, initial shortening velocity and the extent of shortening were reduced, whereas the rate of relaxation was increased. The shape of the curve relating stiffness to isotonic shortening lengths was unchanged, despite the step-induced changes in muscle contractility. Both sets of findings were analyzed in the context of a quasi-structural model describing the shortening-dependent stiffness of lightly loaded tracheal muscle strips. PMID- 15333614 TI - Changes in inorganic phosphate and force production in human skeletal muscle after cast immobilization. AB - Cast immobilization is associated with decreases in muscle contractile area, specific force, and functional ability. The pathophysiological processes underlying the loss of specific force production as well as the role of metabolic alterations are not well understood. The aim of this study was to quantify changes in the resting energy-rich phosphate content and specific force production after immobilization. (31)P-magnetic resonance spectroscopy, three dimensional magnetic resonance imaging, and isometric strength testing were performed in healthy subjects and patients with an ankle fracture after 7 wk of immobilization and during rehabilitation. Muscle biopsies were obtained in a subset of patients. After immobilization, there was a significant decrease in the specific plantar flexor torque and a significant increase in the inorganic phosphate (P(i)) concentration (P < 0.001) and the P(i)-to-phosphocreatine (PCr) ratio (P < 0.001). No significant change in the PCr content or basal pH was noted. During rehabilitation, both the P(i) content and the P(i)-to-PCr ratio decreased and specific torque increased, approaching control values after 10 wk of rehabilitation. Regression analysis showed an inverse relationship between the in vivo P(i) concentration and specific torque (r = 0.65, P < 0.01). In vitro force mechanics performed on skinned human muscle fibers demonstrated that varying the P(i) levels within the ranges observed across individuals in vivo (4 10 mM) changed force production by approximately 16%. In summary, our findings clearly depict a change in the resting energy-rich phosphate content of skeletal muscle with immobilization, which may negatively impact its force generation. PMID- 15333615 TI - Inspiratory flow in the nose: a model coupling flow and vasoerectile tissue distensibility. AB - We have developed a discrete multisegmental model describing the coupling between inspiratory flow and nasal wall distensibility. This model is composed of 14 individualized compliant elements, each with its own relationship between cross sectional area and transmural pressure. Conceptually, this model is based on flow limitation induced by the narrowing of duct due to collapsing pressure. For a given inspiratory pressure and for a given compliance distribution, this model predicts the area profile and inspiratory flow. Acoustic rhinometry and posterior rhinomanometry were used to determine the initial geometric area and mechanical characteristics of each element. The proposed model, used under steady-state conditions, is able to simulate the pressure-flow relationship observed in vivo under normal conditions (4 subjects) and under pathological conditions (4 vasomotor rhinitis and 3 valve syndrome subjects). Our results suggest that nasal wall compliance is an essential parameter to understand the nasal inspiratory flow limitation phenomenon and the associated increase of resistance that is well known to physiologists. By predicting the functional pressure-flow relationship, this model could be a useful tool for the clinician to evaluate the potential effects of treatments. PMID- 15333616 TI - Determinants of fat oxidation during exercise in healthy men and women: a cross sectional study. AB - The aim of the present study was to establish fat oxidation rates over a range of exercise intensities in a large group of healthy men and women. It was hypothesised that exercise intensity is of primary importance to the regulation of fat oxidation and that gender, body composition, physical activity level, and training status are secondary and can explain part of the observed interindividual variation. For this purpose, 300 healthy men and women (157 men and 143 women) performed an incremental exercise test to exhaustion on a treadmill [adapted from a previous protocol (Achten J, Venables MC, and Jeukendrup AE. Metabolism 52: 747-752, 2003)]. Substrate oxidation was determined using indirect calorimetry. For each individual, maximal fat oxidation (MFO) and the intensity at which MFO occurred (Fat(max)) were determined. On average, MFO was 7.8 +/- 0.13 mg.kg fat-free mass (FFM)(-1).min(-1) and occurred at 48.3 +/- 0.9% maximal oxygen uptake (Vo(2 max)), equivalent to 61.5 +/- 0.6% maximal heart rate. MFO (7.4 +/- 0.2 vs. 8.3 +/- 0.2 mg.kg.FFM(-1).min(-1); P < 0.01) and Fat(max) (45 +/- 1 vs. 52 +/- 1% Vo(2 max); P < 0.01) were significantly lower in men compared with women. When corrected for FFM, MFO was predicted by physical activity (self-reported physical activity level), Vo(2 max), and gender (R(2) = 0.12) but not with fat mass. Men compared with women had lower rates of fat oxidation and an earlier shift to using carbohydrate as the dominant fuel. Physical activity, Vo(2 max), and gender explained only 12% of the interindividual variation in MFO during exercise, whereas body fatness was not a predictor. The interindividual variation in fat oxidation remains largely unexplained. PMID- 15333617 TI - Correlated measurement error--implications for nutritional epidemiology. AB - BACKGROUND: In nutritional epidemiology, it is common to fit models in which several dietary variables are included. However, with standard instruments for dietary assessment, not only are the intakes of many nutrients often highly correlated, but the errors in the estimation of the intake of different nutrients are also correlated. The effect of this error correlation on the results of observational studies has been little investigated. This paper describes the effect on multivariate regression coefficients of different levels of correlation, both between the variables themselves and between the errors of estimation of these variables. METHODS: Using a simple model for the multivariate error structure, we examine the effect on the estimates of bivariate linear regression coefficients of (1) differential precision of measurement of the two independent variables, (2) differing levels of correlation between the true values of the two variables, and (3) differing levels of correlation between the errors of measurement of the two variables. As an example, the prediction of plasma vitamin C levels by dietary intake variables is considered, using data from the European Prospective Investigation of Cancer (EPIC) Norfolk study in which dietary intake was estimated using both a food frequency questionnaire (FFQ) and a 7-day diary (7DD). The dietary variables considered are vitamin C, fat, and energy, with different approaches taken to energy adjustment. RESULTS: When the error correlation is zero, the estimates of the bivariate regression coefficients reflect the precision of measurement of the two variables and mutual confounding. The sum of the observed regression coefficients is biased towards the null as in univariate regression. When the error correlation is non-zero but below about 0.7, the effect is minor. However, as the error correlation increases beyond 0.8 the effect becomes large and highly dependent on the relative precision with which the two variables are measured. At the extreme, the bivariate estimates can become indefinitely large. In the example, the error correlation between fat and energy using the FFQ appears to be over 0.9, the corresponding value for the 7DD being approximately 0.85. The error correlation between vitamin C and fat, and vitamin C and energy, appears to be below 0.5 and smaller for the 7DD than for the FFQ. The impact of these error correlations on bivariate regression coefficients is large. The effect of energy adjustment differs widely between vitamin C and fat. CONCLUSION: High levels of error correlation can have a large effect on bivariate regression estimates, varying widely depending on which two variables are considered. In particular, the effect of energy adjustment will vary widely. For vitamin C, the effect of energy adjustment appears negligible, whereas for fat the effect is large indicating that error correlation close to one can partially remove regression dilution due to measurement error. If, for fat intake, energy adjustment is performed by using energy density, the partial removal of regression dilution is achieved at the expense of substantial reduction in the true variance. PMID- 15333618 TI - Adjusting for energy intake--what measure to use in nutritional epidemiological studies? AB - BACKGROUND: The measurement of energy intake in epidemiological studies is difficult. However, it is important that energy intake is assessed if epidemiological analyses are to correspond to isocaloric experiments. The aim of this study was to compare self-reported energy intake, physical activity, and body weight with energy expenditure measured by 4 days of heart rate monitoring with individual calibration of the relationship between heart rate and oxygen consumption. METHODS: Volunteer sub-study of 97 men and women (mean ages 54 and 51 years respectively) within the European Investigation into Cancer (EPIC) study in Norfolk (UK). Dietary assessment of energy intake and physical activity was by self-report and weight was measured using standard techniques. Energy expenditure was assessed objectively by recording heart rate for 4 days following a calibration of the relationship between heart rate and oxygen consumption. RESULTS: Self-reported energy intake by 7-day diary (mean 8.5 MJ/day) and food frequency questionnaire (FFQ) (mean 8.8 MJ/day) were significantly lower than objectively measured total energy expenditure (mean 11.2 MJ/day). The deattenuated partial correlations between total energy expenditure were 0.33 (7 day diary), 0.34 (FFQ), 0.50 (physical activity), and 0.56 (weight). Weight accounted for 31% (deattenuated) of the sum of squares about the mean of true energy intake after adjusting for age and sex. With the addition of self-reported physical activity, the model was significantly improved (R2 = 0.57). Adding energy either assessed by the diary or FFQ did not improve the model. CONCLUSIONS: The results presented here indicate that to adjust for energy intake, for the purpose of replicating an isocaloric experiment in an observational epidemiological study, one would do considerably better adjusting for weight and physical activity, than adjusting for energy intake estimated from an FFQ. PMID- 15333619 TI - Comparison of imputation and modelling methods in the analysis of a physical activity trial with missing outcomes. AB - BACKGROUND: Longitudinal studies almost always have some individuals with missing outcomes. Inappropriate handling of the missing data in the analysis can result in misleading conclusions. Here we review a wide range of methods to handle missing outcomes in single and repeated measures data and discuss which methods are most appropriate. METHODS: Using data from a randomized controlled trial to compare two interventions for increasing physical activity, we compare complete case analysis; ad hoc imputation techniques such as last observation carried forward and worst-case; model-based imputation; longitudinal models with random effects; and recently proposed joint models for repeated measures data and non ignorable dropout. RESULTS: Estimated intervention effects from ad hoc imputation methods vary widely. Standard multiple imputation and longitudinal modelling agree closely, as they should. Modifying the modelling method to allow for non ignorable dropout had little effect on estimated intervention effects, but imputing using a common imputation model in both groups gave more conservative results. CONCLUSIONS: Results from ad hoc imputation methods should be avoided in favour of methods with more plausible assumptions although they may be computationally more complex. Although standard multiple imputation methods and longitudinal modelling methods are equivalent for estimating the treatment effect, the two approaches suggest different ways of relaxing the assumptions, and the choice between them depends on contextual knowledge. PMID- 15333620 TI - Prevalence, awareness and treatment of hypercholesterolaemia in 32 populations: results from the WHO MONICA Project. AB - BACKGROUND: Several studies have been conducted to estimate the population prevalence of hypertension, or its diagnosis and treatment. There is no multinationally comparable information on the prevalence of hypercholesterolaemia, or its diagnosis and treatment, since individual studies are often not directly comparable. METHODS: Data from the WHO MONICA Project's final risk factor surveys were used. Data were collected using standardized methods between 1989 and 1997 for the 35-64 year age range in 32 populations, in 19 countries on 3 continents. RESULTS: The prevalence of hypercholesterolaemia (total cholesterol > or = 6.5 mmol/l or taking lipid-lowering drugs) varied across populations from 3% to 53% in men, and from 4% to 40% in women. Awareness of hypercholesterolaemia varied from 1% to 33% in men, and from 0% to 31% in women. In most populations, over 50% of men and women on lipid-lowering drugs had a cholesterol level < 6.5 mmol/l. CONCLUSIONS: There is wide variation in the prevalence, awareness, and treatment of hypercholesterolaemia between populations. For the planning and implementation of primary prevention programmes and for the development of health care systems, monitoring of changes, both within and between populations, is essential. To obtain reliable information on these changes, well-standardized methods must be applied. PMID- 15333621 TI - Regression to the mean: what it is and how to deal with it. AB - BACKGROUND: Regression to the mean (RTM) is a statistical phenomenon that can make natural variation in repeated data look like real change. It happens when unusually large or small measurements tend to be followed by measurements that are closer to the mean. METHODS: We give some examples of the phenomenon, and discuss methods to overcome it at the design and analysis stages of a study. RESULTS: The effect of RTM in a sample becomes more noticeable with increasing measurement error and when follow-up measurements are only examined on a sub sample selected using a baseline value. CONCLUSIONS: RTM is a ubiquitous phenomenon in repeated data and should always be considered as a possible cause of an observed change. Its effect can be alleviated through better study design and use of suitable statistical methods. PMID- 15333622 TI - How far are socioeconomic differences in coronary heart disease hospitalization, all-cause mortality and cardiovascular mortality among adult Swedish males attributable to negative childhood circumstances and behaviour in adolescence? AB - OBJECTIVES: Coronary heart disease (CHD) and mortality are unevenly distributed between social classes, the lower being disadvantaged compared with the higher. Adverse social circumstances in childhood have been related to an increased risk of CHD and mortality in adulthood. The present purpose was to investigate the effect of differences in social disadvantage in early life on later differences in risk of CHD, all-cause mortality, and cardiovascular mortality between socioeconomic groups among Swedish men aged 40-50. METHODS: Data on circumstances in childhood and adolescence, e.g. crowded housing and low social position of the father, measured at age 9-11, was collected among 49,323 men, born in 1949-51, and conscripted for compulsory military training in 1969/70. Data on adult socioeconomic position was obtained from the 1990 census, data on CHD from the Swedish In-patient Care register 1991-2000, and data on mortality from the Causes of Death register 1991-2000. RESULTS: A social gradient was found for all health outcomes. The relative risk of CHD, all-cause mortality, and cardiovascular mortality among unskilled workers compared with that among high-level non-manual employees was 1.82 (95% CI: 1.36, 2.44), 2.24 (95% CI: 1.72, 2.93), and 2.38 (95% CI: 1.47, 3.86) respectively. The early life risk indicators, such as crowded housing and low childhood social position (measured at age 9-11), short stature (measured at age 18-20), and low education (reported at age 18-20), were more common among those who in 1990 (i.e. at age 39-41) were manual workers than among those who were in non-manual occupations. In multivariate analyses, considering the indicators of childhood social disadvantage and adjusting for lifestyle factors established at age 18-20, (smoking, alcohol consumption, overweight) the increased relative risk of CHD hospitalization and cardiovascular mortality in the four categories of employed workers was reduced by 72-100%. CONCLUSION: Predictors of CHD measured in childhood and adolescence may explain a substantial part of the social gradient in CHD, cardiovascular mortality, and all-cause mortality among the 40-50 year old males studied. PMID- 15333623 TI - Commentary: Correlated errors and energy adjustment--where are the data? PMID- 15333624 TI - Collaboration is needed to co-ordinate European birth cohort studies. PMID- 15333625 TI - Medial vestibular nucleus mediates the cardiorespiratory responses to fastigial nuclear activation and hypercapnia. AB - Electrical stimulation of the cerebellar fastigial nucleus (FN) evokes hyperventilation and hypertension responses that are similar to those induced by stimulation of the medial region of the vestibular nucleus (VNM). Because there are mutual projections between these two nuclei morphologically, we hypothesized that the FN-mediated cardiorespiratory responses were related to the integrity of the VNM. Experiments were conducted on 21 anesthetized, tracheotomized, and spontaneously breathing rats. Electrical stimulation (approximately 10 s) of the FN was used to evoke cardiorespiratory responses, and the same stimulus was repeated 30-45 min after bilateral lesions of the VNM by local microinjection of ibotenic acid (100 mM, 100 nl). We found that FN stimulation-induced hyperventilation and hypertension were attenuated significantly by the lesions. The role of the VNM in the ventilatory responses to chemical challenges was subsequently defined. The animals were exposed to hypercapnia (10% CO2) and hypoxia (10% O2) for 1-2 min randomly before and after VNM lesions. The results showed that VNM lesions significantly attenuated the cardiorespiratory responses to hypercapnia but not to hypoxia, with little effect on baseline respiratory variables. These findings suggest that the VNM is required for full expression of the cardiorespiratory responses to electrical stimulation of the FN as well as to hypercapnia. However, neurons within the VNM do not appear to be critical for maintaining eupneic breathing and the cardiorespiratory responses to hypoxia. PMID- 15333626 TI - Strain differences in murine ventilatory behavior persist after urethane anesthesia. AB - Differences in breathing pattern between awake C57BL/6J (B6) and A/J mice are such that A/J mice breathe slower, deeper, and with greater variability than B6. We theorized that urethane anesthesia, by affecting cortical and subcortical function, would test the hypothesis that strain differences require a fully functional neuroaxis. We anesthetized B6 and A/J mice with urethane, placed them in a whole-body plethysmograph, and measured the durations of inspiration and expiration, respiratory frequency (Fr), and peak amplitude during exposure to room air (21% O2), hyperoxia (5 min, 100% O2), hypoxia (5 min, 8% O2), and posthypoxic reoxygenation (5 min, 100% O2). Breathing variability was assessed by calculating the coefficient of variation (CV) and by applying spatial statistics to Poincare plots constructed from the timing and amplitude data. Even though Fr in anesthetized B6 and A/J mice was greater than that for unanesthetized animals, anesthetized A/J mice still breathed slower, deeper, and with greater variability than B6 mice at rest and during hyperoxia. During the fourth minute of hypoxia, Fr and its CV were not significantly different between strains. Even though Fr was similar between strains immediately after hypoxia, its CV was significantly greater for B6 than A/J mice. Posthypoxic Fr was significantly less than baseline Fr in B6 but not A/J mice, and the CV for posthypoxic Fr was greater for B6 but less for AJ mice compared with baseline CV. This difference in patterning was confirmed by spatial statistical analysis. We conclude that strain-specific differences in respiratory pattern and its variability are robust genetic traits. The neural substrate for these differences, at least partially, exists within subcortical structures generating the breathing pattern. PMID- 15333627 TI - Continuous measurement of gas uptake and elimination in anesthetized patients using an extractable marker gas. AB - Measurement of pulmonary gas uptake and elimination is often performed, using nitrogen as marker gas to measure gas flow, by applying the Haldane transformation. Because of the inability to measure nitrogen with conventional equipment, measurement is difficult during inhalational anesthesia. A new method is described, which is compatible with any inspired gas mixture, in which fresh gas and exhaust gas flows are measured using carbon dioxide as an extractable marker gas. A system was tested in eight patients undergoing colonic surgery for automated measurement of uptake of oxygen, nitrous oxide, isoflurane, and elimination of carbon dioxide with this method. Its accuracy and precision were compared with simultaneous measurements made with the Haldane transformation and corrected for predicted nitrogen excretion by the lungs. Good agreement was obtained for measurement of uptake or elimination of all gases studied. Mean bias was -0.003 l/min for both oxygen and nitrous oxide uptake, -0.0002 l/min for isoflurane uptake, and 0.003 l/min for carbon dioxide elimination. Limits of agreement lay within 30% of the mean uptake rate for nitrous oxide, within 15% for oxygen, within 10% for isoflurane, and within 5% for carbon dioxide. The extractable marker gas method allows accurate and continuous measurement of gas uptake and elimination in an anesthetic breathing system with any inspired gas mixture. PMID- 15333628 TI - Modulation of microvascular function following low-dose exposure to the organophosphorous compound malathion in human skin in vivo. AB - This study investigates whether malathion, a widely used organophosphate insecticide, has its effects on cutaneous vasculature in healthy human volunteers through its anticholinergic activity or through the modulation of other, noncholinergic pathways. Acute, low-dose exposure to malathion (10 mg/ml for 5 h under occlusive dressing) caused a significant increase in cutaneous blood flux, monitored by using laser-Doppler flowmetry and imaging. It had little effect on tissue levels of ACh, nitric oxide, and histamine assayed in dermal dialysate collected from malathion-exposed and control-treated skin. The duration of the cutaneous vascular response to exogenous ACh (2%) delivered by iontophoresis was significantly enhanced by preexposure to malathion, both <1 h after its removal and 24 h later (P < 0.001). At <1 h, the time to 50% decay of the response was 24 +/- 4 and 50 +/- 8 min in control and malathion-treated skin, respectively. Malathion also enhanced the size and duration of the axon reflex-mediated vasoresponse to ACh. The increase in blood flux to malathion and the endothelium mediated response to exogenous ACh, both in the presence and absence of malathion, were attenuated by pretreatment of the skin with atropine and local anesthesia (P < 0.01). We conclude that short-term exposure to a single low dose of malathion causes prolonged modulation of the physiological function of the cutaneous vasculature and that this is, in part, through its action on acetylcholinesterase at both neuronal and nonneuronal sites. PMID- 15333629 TI - Integrative control of the skeletal muscle microcirculation in the maintenance of arterial pressure during exercise. AB - Skeletal muscle blood flow and vascular conductance are influenced by numerous factors that can be divided into two general categories: central cardiovascular control mechanisms and local vascular control mechanisms. Central cardiovascular control mechanisms are thought to be designed primarily for the maintenance of arterial pressure and central cardiovascular homeostasis, whereas local vascular control mechanisms are thought to be designed primarily for the maintenance of muscle homeostasis. To support the high metabolic rates that can be generated during muscle contraction, skeletal muscle has a tremendous capacity to vasodilate and increase oxygen and nutrient delivery. During whole body dynamic exercise at maximal oxygen consumption (VO2 max), the skeletal muscle receives 85 90% of cardiac output. Yet despite receiving such a large fraction of cardiac output during high-intensity exercise, a vasodilator reserve remains with the potential to produce further elevations in skeletal muscle vascular conductance and blood flow. However, because maximal cardiac output is reached during exercise at VO2 max, further elevations in muscle vascular conductance would produce a fall in arterial pressure. Therefore, limits on muscle perfusion must be imposed during whole body exercise to prevent such drops in pressure. Effective arterial pressure control in response to a potentially hypotensive challenge during high-intensity exercise occurs primarily through reflex-mediated increases in sympathetic nerve activity, which are capable of modulating vasomotor tone of the skeletal muscle resistance vasculature. Thus skeletal muscle vascular conductance and perfusion are primarily mediated by local factors at rest and during exercise, but other centrally mediated control systems are superimposed on the dominant local control mechanisms to provide an integrated regulation of both arterial pressure and skeletal muscle vascular conductance and perfusion during whole body dynamic exercise. PMID- 15333630 TI - What makes vessels grow with exercise training? AB - Exercise and muscle contractions create a powerful stimulus for structural remodeling of the vasculature. An increase in flow velocity through a vessel increases shear stress, a major stimulus for enlargement of conduit vessels. This leads to an endothelial-dependent, nitric oxide-dependent enlargement of the vessel. Increased flow within muscle, in the absence of contractions, leads to an enhanced capillarity by intussusceptive angiogenesis, a process of capillary splitting by intraluminal longitudinal divide. In contrast, sprouting angiogenesis requires extensive endothelial cell proliferation, with degradation of the extracellular matrix to permit migration and tube formation. This occurs during muscle adaptations to chronic contractions and/or muscle overload. The angiogenic growth factor VEGF appears to be an important element in angiogenesis. Recent advances in research have identified hemodynamic and mechanical stimuli that upregulate angiogenic processes, demonstrated a complexity of potent growth factors and interactions with their corresponding receptors, detected an interaction of cellular signaling events, and identified important tissue reorganization processes that must be coordinated to effect vascular remodeling. It is likely that much of this information is applicable to the vascular remodeling that occurs in response to exercise and/or muscle contractions. PMID- 15333632 TI - Insights into the role of an active site aspartate in Ty1 reverse transcriptase polymerization. AB - Long terminal repeat-containing retrotransposons encode reverse transcriptases (RTs) that replicate their RNA into integratable, double-stranded DNA. A mutant version of the RT from Saccharomyces cerevisiae retrotransposon Ty1, in which one of the three active site aspartates has been changed to asparagine (D211N), is still capable of in vitro polymerization, although it is blocked for in vivo transposition. We generated recombinant WT and D211N Ty1 RTs to study RT function and determine specific roles for the Asp(211) residue. Presteady-state kinetic analysis of the two enzymes shows that the D211N mutation has minimal effect on nucleotide binding but reduces the k(pol) by approximately 230-fold. The mutation reduces binding affinity for both Mn(2+) and Mg(2+), indicating that the Asp(211) side chain helps create a tight metal binding pocket. Although both enzymes are highly processive and tend to remain bound to their initial substrate, each shows distinctive patterns of pausing, attributable to interactions between metal ions and the active site residue. These results provide insights to specific roles for the Asp(211) residue during polymerization and indicate unusual enzymatic properties that bear on the Ty1 replication pathway. PMID- 15333633 TI - p21-activated kinase regulates endothelial permeability through modulation of contractility. AB - Endothelial cells lining the vasculature have close cell-cell associations that maintain separation of the blood fluid compartment from surrounding tissues. Permeability is regulated by a variety of growth factors and cytokines and plays a role in numerous physiological and pathological processes. We examined a potential role for the p21-activated kinase (PAK) in the regulation of vascular permeability. In both bovine aortic and human umbilical vein endothelial cells, PAK is phosphorylated on Ser141 during the activation downstream of Rac, and the phosphorylated subfraction translocates to endothelial cell-cell junctions in response to serum, VEGF, bFGF, TNFalpha, histamine, and thrombin. Blocking PAK activation or translocation prevents the increase in permeability across the cell monolayer in response to these factors. Permeability correlates with myosin phosphorylation, formation of actin stress fibers, and the appearance of paracellular pores. Inhibition of myosin phosphorylation blocks the increase in permeability. These data suggest that PAK is a central regulator of endothelial permeability induced by multiple growth factors and cytokines via an effect on cell contractility. PAK may therefore be a suitable drug target for the treatment of pathological conditions where vascular leak is a contributing factor, such as ischemia and inflammation. PMID- 15333634 TI - An RNA ligase from Deinococcus radiodurans. AB - Although DNA repair pathways have been the focus of much attention, there is an emerging appreciation that distinct pathways exist to maintain or manipulate RNA structure in response to breakage events. Here we identify an RNA ligase (DraRnl) from the radiation-resistant bacterium Deinococcus radiodurans. DraRnl seals 3' OH/5'-PO4 RNA nicks in either a duplex RNA or an RNA: DNA hybrid, but it cannot seal 3'-OH/5'-PO4 DNA nicks. The specificity of DraRnl arises from a requirement for RNA on the 3'-OH side of the nick. DraRnl is a 342-amino acid monomeric protein with a distinctive structure composed of a C-terminal adenylyltransferase domain linked to an N-terminal module that resembles the OB-fold of phenylalanyl tRNA synthetases. RNA sealing activity was abolished by mutation of the predicted lysine adenylylation site (Lys-165) in the C-terminal domain and was reduced by an order of magnitude by deletion of the N-terminal OB module. Our findings highlight the existence of an RNA repair capacity in bacteria and support the hypothesis that contemporary DNA ligases, RNA ligases, and RNA capping enzymes evolved by the fusion of ancillary effector domains to an ancestral catalytic module involved in RNA repair. PMID- 15333635 TI - Promiscuous target interactions in the mariner transposon Himar1. AB - We have previously characterized the early intermediates of mariner transposition. Here we characterize the target interactions that occur later in the reaction. We find that, in contrast to the early transposition intermediates, the strand transfer complex is extremely stable and difficult to disassemble. Transposase is tightly bound to the transposon ends constraining rotation of the DNA at the single strand gaps in the target site flanking the element on either side. We also find that although the cleavage step requires Mg2+ or Mn2+ as cofactor, the strand transfer step is also supported by Ca2+, suggesting that the structure of the active site changes between cleavage and insertion. Finally, we show that, in contrast to the bacterial cut and paste transposons, mariner target interactions are promiscuous and can take place either before or after cleavage of the flanking DNA. This is similar to the behavior of the V(D)J system, which is believed to be derived from an ancestral eukaryotic transposon. We discuss the implications of promiscuous target interactions for promoting local transposition and whether this is an adaptation to facilitate the invasion of a genome following horizontal transfer to a new host species. PMID- 15333636 TI - Role of Lys-32 residues in R67 dihydrofolate reductase probed by asymmetric mutations. AB - R67 dihydrofolate reductase (R67 DHFR) is a novel protein encoded by an R-plasmid that confers resistance to the antibiotic, trimethoprim. This homotetrameric enzyme possesses 222 symmetry, which imposes numerous constraints on the single active site pore, including a "one-site-fits-both" strategy for binding its ligands, dihydrofolate (DHF) and NADPH. Previous studies uncovered salt effects on binding and catalysis (Hicks, S. N., Smiley, R. D., Hamilton, J. B., and Howell, E. E. (2003) Biochemistry 42, 10569-10578), however the one or more residues that participate in ionic contacts with the negatively charged tail of DHF as well as the phosphate groups in NADPH were not identified. Several studies predict that Lys-32 residues were involved, however mutations at this residue destabilize the R67 DHFR homotetramer. To study the role of Lys-32 in binding and catalysis, asymmetric K32M mutations have been utilized. To create asymmetry, individual mutations were added to a tandem array of four in-frame gene copies. These studies show one K32M mutation is tolerated quite well, whereas addition of two mutations has variable effects. Two double mutants, K32M:1+2 and K32M: 1+4, which place the mutations on opposite sides of the pore, reduce kcat. However a third double mutant, K32M: 1+3, that places two mutations on the same half pore, enhances kcat 4- to 5-fold compared with the parent enzyme, albeit at the expense of weaker binding of ligands. Because the kcat/Km values for this double mutant series are similar, these mutations appear to have uncovered some degree of non productive binding. This non-productive binding mode likely arises from formation of an ionic interaction that must be broken to allow access to the transition state. The K32M:1+3 mutant data suggest this interaction is an ionic interaction between Lys-32 and the charged tail of dihydrofolate. This unusual catalytic scenario arises from the 222 symmetry imposed on the single active site pore. PMID- 15333637 TI - "Catch 222," the effects of symmetry on ligand binding and catalysis in R67 dihydrofolate reductase as determined by mutations at Tyr-69. AB - R67 dihydrofolate reductase (R67 DHFR) catalyzes the transfer of a hydride ion from NADPH to dihydrofolate, generating tetrahydrofolate. The homotetrameric enzyme provides a unique environment for catalysis as both ligands bind within a single active site pore possessing 222 symmetry. Mutation of one active site residue results in concurrent mutation of three additional symmetry-related residues, causing large effects on binding of both ligands as well as catalysis. For example, mutation of symmetry-related tyrosine 69 residues to phenylalanine (Y69F), results in large increases in Km values for both ligands and a 2-fold rise in the kcat value for the reaction (Strader, M. B., Smiley, R. D., Stinnett, L. G., VerBerkmoes, N. C., and Howell, E. E. (2001) Biochemistry 40, 11344 11352). To understand the interactions between specific Tyr-69 residues and each ligand, asymmetric Y69F mutants were generated that contain one to four Y69F mutations. A general trend observed from isothermal titration calorimetry and steady-state kinetic studies of these asymmetric mutants is that increasing the number of Y69F mutations results in an increase in the Kd and Km values. In addition, a comparison of steady-state kinetic values suggests that two Tyr-69 residues in one half of the active site pore are necessary for NADPH to exhibit a wild-type Km value. A tyrosine 69 to leucine mutant was also generated to approach the type(s) of interaction(s) occurring between Tyr-69 residues and the ligands. These studies suggest that the hydroxyl group of Tyr-69 is important for interactions with NADPH, whereas both the hydroxyl group and hydrophobic ring atoms of the Tyr-69 residues are necessary for proper interactions with dihydrofolate. PMID- 15333638 TI - Activation of endothelial nitric-oxide synthase by the p38 MAPK in response to black tea polyphenols. AB - Black tea improves endothelial function in patients with coronary artery disease. We sought to determine the responsible components of black tea and elucidate the underlying cell signaling mechanisms. We exposed porcine aortic endothelial cells to components of black tea and found that the polyphenol fraction acutely enhanced nitric oxide bioactivity. This effect involved endothelial nitric-oxide synthase (eNOS) phosphorylation at Ser-1177 and dephosphorylation at Thr-495, consistent with increased eNOS activity. These effects were calcium-dependent, as removal of extracellular calcium prevented eNOS phosphorylation at Ser-1177, whereas inhibition of intracellular calcium mobilization with TMB-8 blunted Thr 495 dephosphorylation. Black tea polyphenol-induced eNOS activation appeared dependent upon the phosphatidylinositol 3-kinase-Akt pathway, as it was significantly inhibited by LY294002 and a dominant negative Akt, respectively. Pharmacological inhibition of p38 mitogen-activated protein kinase (p38 MAPK) with either SB202190 or SB203580 as well as overexpression of a dominant negative p38 MAPKalpha attenuated both eNOS activation and phosphorylation changes in response to black tea polyphenols. Inhibition of p38 MAPKalpha also blunted Akt activation in response to black tea polyphenols, suggesting that p38alpha MAPK is upstream of Akt in this pathway. Finally, a constitutively active mutant of MKK6bE, an upstream kinase for p38 MAPK, enhanced both the basal and stimulated activity of Akt, leading to increased eNOS activity. Taken together, these data identify the p38 MAPK as an upstream component of Akt-mediated eNOS activation. PMID- 15333639 TI - The central role of the green alga Chlamydomonas reinhardtii in revealing the mechanism of state transitions. AB - This review focuses on the essential role played by the green alga Chlamydomonas reinhardtii in revealing both the mechanism and the physiological consequences of state transitions. Two aspects are considered. The first is the role of the cytochrome b6f complex in regulating state transitions, in light of the recently obtained 3D structure. The second is the switch between linear and cyclic electron flow that follows state transitions in Chlamydomonas. Structural and dynamic elements that might be involved in such a switch, as well as its consequences on the energetic metabolism, are discussed. PMID- 15333640 TI - Cloning and characterization of a 2-Cys peroxiredoxin from Pisum sativum. AB - A cDNA sequence coding for a pea (Pisum sativum L.) 2-Cys peroxiredoxin (2-Cys Prx) has been cloned. The deduced amino acid sequence showed a high sequence homology to the 2-Cys Prx enzymes of Phaseolus vulgaris (86%), Arabidopsis thaliana (75%), and Spinacia oleracea (75%), and contained a chloroplast target sequence at its N-terminus. The mature enzyme, without the transit peptide, has a molecular mass of 22 kDa as well as two cysteine residues (Cys-53 and Cys-175) which are well conserved among proteins of this group. The protein was expressed in a heterologous system using the expression vector pET3d, and was purified to homogeneity by three sequential chromatographic steps. The enzyme exhibits peroxidase activity on hydrogen peroxide (H(2)O(2)) and t-butyl hydroperoxide (TBHP) with DTT as reducing agent. Although both pea Trxs f and m reduce oxidized 2-Cys Prx, Trx m is more efficient. The precise conditions for oligomerization of 2-Cys Prx through extensive gel filtration studies are also reported. The transition dimer-decamer produced in vitro between pH 7.5 and 8.0 and the influence of DTT suggest that a great change in the enzyme quaternary structure of 2-Cys Prx may take place in the chloroplast during the dark-light transition. In addition, the cyclophilin-dependent reduction of chloroplast 2-Cys Prx is shown. PMID- 15333641 TI - Maize ribosome-inactivating proteins (RIPs) with distinct expression patterns have similar requirements for proenzyme activation. AB - Ribosome-inactivating proteins (RIPs, EC 3.2.2.22) are potent naturally occurring toxins found in numerous and diverse plant species. The maize RIP is unusual among the plant RIPs because it is synthesized as an inactive precursor (also known as maize proRIP1 or b-32). The proenzyme undergoes proteolytic activation that results in the removal of the NH(2)-terminal, the COOH-terminal, and internal sequences to form a two-chain holoenzyme capable of irreversibly modifying the large rRNA. The characterization of a second maize RIP (RIP2), encoded by the gene designated Rip3:2 is described here. Low levels of Rip3:2 RNA were detected in roots, shoots, tassels, silks, and leaves, but the Rip3:2 gene, unlike the Rip3:1 gene, is not under the control of the transcriptional activator Opaque-2. Instead, its expression was up-regulated by drought. Rip3:2 encodes a 31.1 kDa polypeptide that is very similar to proRIP1 in regions corresponding to those found in the active protein and the NH(2)-terminal extension. A 19-amino acid internal portion of proRIP2 has little similarity to the proRIP1 sequence except that both are very rich in acidic residues. RIP activity assays revealed that Rip3:2 encodes a polypeptide that acquires RNA-specific N-glycosidase activity after proteolytic cleavage. Accumulation as inactive proenzymes may therefore be a general feature of maize RIPs. Differential regulation of the two RIP genes suggests that the corresponding proteins may be involved in defence related functions with one being regulated developmentally and the other being responsive to an environmental stimulus. PMID- 15333642 TI - Involvement of ethylene signalling in a non-climacteric fruit: new elements regarding the regulation of ADH expression in grapevine. AB - Although grape berries have been classified as non-climacteric fruits, ongoing studies on grape ethylene signalling challenge the role of ethylene in their ripening. One of the significant molecular changes in berries is the up regulation of ADH (alcohol dehydrogenase, EC 1.1.1.1) enzyme activity at the inception of fruit ripening and of VvADH2 transcript levels. This paper shows that the ethylene signal transduction pathway could be involved in the control of VvADH2 expression in grapevine berries and in cell suspensions. The induction of VvADH2 transcription, either in berries at the inception of ripening or in cell suspensions, was found to be partly inhibited by 1-methylcyclopropene (1-MCP), an inhibitor of ethylene receptors. Treatment of cell suspensions with 2 chloroethylphosphonic acid (2-CEPA), an ethylene-releasing compound, also resulted in a significant increase in ADH activity and VvADH2 transcription under anaerobiosis, showing that concomitant ethylene and anaerobic treatments in cell suspensions could result in changes in VvADH2 expression. All these results associated with the presence in the VvADH2 promoter of regulatory elements for ethylene and anaerobic response, suggest that the ethylene transduction pathway and anaerobic stress could be, in part, involved in the regulation of VvADH2 expression in ripening berries and cell suspensions. These data open new aspects of the expression control of a ripening-related gene in a non-climacteric fruit. PMID- 15333643 TI - Flower and fruit abortion in sweet pepper in relation to source and sink strength. AB - Source strength (assimilate supply) and sink strength (assimilate demand) of the plant were varied in different ways to investigate to what extent flower/fruit abortion in sweet pepper (Capsicum annuum L.) is determined by the availability of assimilates. Source strength was varied by changing the light level, plant density, and leaf pruning. Sink strength was varied by changing the temperature and the number and position of earlier formed fruits. Shading as well as heating for short periods showed that flowers/fruits were the most susceptible to abortion during the first week after anthesis. The different experiments where source strength was varied all showed that when source strength decreased, the rate of abortion increased linearly, whether source strength was decreased by shading, high plant density, or leaf pruning. That flower and fruit abortion not only depends on the source strength but also on the sink strength of competing organs is shown by varying the number or the position of earlier formed fruits. With the same source strength, the rate of abortion showed a close relationship with the growth rate of the earlier formed competing fruits, suggesting that the induction of abortion by earlier formed fruits is due to their sink strength. Most of the variation in abortion could be related to differences in vegetative growth rate, the latter being an indicator of the source-sink ratio. However, with the same vegetative growth rate, the rate of abortion was lower for the leaf pruning treatments where no competing fruits were retained than for the fruit load treatments. This indicates that although most of the variation in abortion can be related to the source and sink strength of the plant, some effects of competing fruits can only be explained by a combination of competition and dominance. PMID- 15333644 TI - Spatial mapping of phosphorus influx in bean root systems using digital autoradiography. AB - A novel technique was developed to spatially map the phosphorus net influx capacity in intact root systems. The method is based on digital autoradiography and permits the quantification of phosphorus influx at high spatial resolution (2 mm). Roots of 18-d-old common bean plants were exposed to (32)P-labelled orthophosphate, quickly frozen, excised, lyophilized, scanned, and exposed to a storage phosphor screen. Plots of (32)P content versus root length (distance from the root tip or from the base of the root) were obtained for three different root classes: basal, basal laterals, and taproot laterals. Radioactivity detected by filmless autoradiography correlated well (r(2)=0.99) with measurements made by scintillation counting. Basal roots absorbed 2.5 times and 1.9 times more phosphorus than the taproot lateral and basal lateral root classes, respectively, in the first 20 mm from the root apex. External phosphorus markedly affected influx: roots averaged 5, 16, and 34 pmol P min(-1) in the apical 20 mm when exposed to 1, 5, and 10 microM P solutions, respectively. The spatial pattern of phosphorus influx along the root axes of the different root classes was rather homogeneous when measured on a root surface area basis. Phosphorus influx in the older segments of basal roots (those next to the hypocotyl) did not differ from the newer segments close to the root apex. However, a heterogeneous pattern was detected for basal roots when measured on a length basis, indicating that both root class and diameter constitute main factors controlling the spatial pattern of net influx. PMID- 15333645 TI - A comparative study of the role of the major proteinases of germinated common bean (Phaseolus vulgaris L.) and soybean (Glycine max (L.) Merrill) seeds in the degradation of their storage proteins. AB - Two types of cysteine proteases, low-specificity enzymes from the papain family and Asn-specific from the legumain family are generally considered to be the major endopeptidases responsible for the degradation of seed storage proteins during early seedling growth. The action of the corresponding enzymes (CPPh1 and LLP, respectively) from common bean (Phaseolus vulgaris L.) on phaseolin (the common bean storage protein), and on the homologous soybean (Glycine max (L.) Merrill) storage protein, beta-conglycinin, was studied. Under the action of LLP, proteolysis of phaseolin was limited to cleavage of its interdomain linker. No cleavage of the interdomain linker occurred in beta-conglycinin with LLP. LLP action was restricted to splitting off the disordered N-terminal extensions of alpha and alpha' subunits. No extensive hydrolysis (degradation to short TCA soluble peptides) of either protein occurred under the action of LLP. CPPh1 cleaved the phaseolin subunits into roughly half-sized fragments at the onset of proteolysis. The cleavage was accompanied by a small (8-10%) decrease of protein. No decrease of protein occurred with further incubation. Thus the two most active proteinases detected in common bean seedlings individually were incapable of the extensive degradation of phaseolin. Extensive hydrolysis of phaseolin was only achieved by the consecutive action of LLP and CPPh1. Similar cleavages occurred during the action of CPPh1 on beta-conglycinin. However, by contrast with phaseolin, CPPh1 by itself accomplished the extensive hydrolysis of beta conglycinin. The differences in the course of proteolysis of the proteins studied were determined by their structural peculiarities. PMID- 15333646 TI - European, Chinese and Japanese pear fruits exhibit differential softening characteristics during ripening. AB - Softening characteristics were investigated in three types of pear fruit, namely, European pear 'La France', Chinese pear 'Yali', and Japanese pear 'Nijisseiki'. 'La France' fruit softened dramatically and developed a melting texture during ripening, while 'Yali' fruit with and without propylene treatment showed no change in flesh firmness and texture during ripening. Non-treated 'Nijisseiki' did not show a detectable decrease in flesh firmness, whereas continuous propylene treatment caused a gradual decrease in firmness resulting in a mealy texture. In 'La France', the analysis of cell wall polysaccharides revealed distinct solubilization and depolymerization of pectin and hemicellulose during fruit softening. In 'Nijisseiki', propylene treatment led to the solubilization and depolymerization of pectic polysaccharides to a limited extent, but not of hemicellulose. In 'Yali', hemicellulose polysaccharides were depolymerized during ripening, but there was hardly any change in pectic polysaccharides except in the water-soluble fraction. PC-PG1 and PC-PG2, two polygalacturonase (PG) genes, were expressed in 'La France' fruit during ripening, while only PC-PG2 was expressed in 'Nijisseiki' and neither PC-PG1 or PC-PG2 was expressed in 'Yali'. The expression pattern of PC-XET1 was constitutive during ripening in all three pear types. PG activity measured by the reducing sugar assay increased in all three pears during ripening. However, viscometric measurements showed that the levels of endo-PG activity were high in 'La France', low in 'Nijisseiki', and undetectable in 'Yali' fruits. These results suggest that, in pears, cell wall degradation is correlated with a decrease in firmness during ripening and the modification of both pectin and hemicellulose are essential for the development of a melting texture. Furthermore, the data suggest that different softening behaviours during ripening among the three pear fruits may be caused by different endo-PG activity and different expression of PG genes. PMID- 15333647 TI - Best Practice No 179. Guidelines for breast needle core biopsy handling and reporting in breast screening assessment. AB - Non-operative diagnosis has become the norm in breast disease assessment and, until relatively recently, fine needle aspiration cytology has been the sampling method of choice. The introduction of automated core biopsy guns in the mid 1990s led to the additional introduction of core biopsy in assessment units. This paper presents a summary of the guidance on handling and routine reporting of breast needle core biopsy specimens in the context of breast disease multidisciplinary assessment. This guidance has been produced by the UK National Coordinating Committee for Breast Screening Pathology and is endorsed by the European Commission working group on breast screening pathology. PMID- 15333648 TI - The detection of circulating breast cancer cells in blood. AB - At present, sampling of the lymph nodes or bone marrow for the detection of regions of metastatic disease in patients with breast cancer can only be undertaken at the time of initial diagnosis and surgery. However, the sampling of these tissues and the methods used are inaccurate, time consuming, and cannot be used for easy routine screening to determine disease recurrence and response to treatment. Because of the problems encountered with current methods and tissues sampled at the time of breast cancer diagnosis, this review discusses the urgent requirement for and potential development of a quick, simple, and accurate diagnostic test utilising the haematogenous system, a source of circulating tumour cells in patients with breast cancer, and highly sensitive molecular biological techniques, such as reverse transcription polymerase chain reaction. In addition, this review also highlights potential problems that may be encountered and should be avoided when devising such a test. PMID- 15333649 TI - Demystified...recombinant antibodies. AB - Recombinant antibodies are important tools for biomedical research and are increasingly being used as clinical diagnostic/therapeutic reagents. In this article, a background to humanized antibodies is given, together with details of the generation of antibody fragments--for example, single chain Fv fragments. Phage antibody fragments are fast becoming popular and can be generated by simple established methods of affinity enrichment from libraries derived from immune cells. Phage display methodology can also be used for the affinity enrichment of existing antibody fragments to provide a reagent with a higher affinity. Here, phage antibodies are demystified to provide a greater understanding of the potential of these reagents and to engage clinicians and biomedical scientists alike to think about potential applications in pathology and clinical settings. PMID- 15333650 TI - Morules in endometrial carcinoma and benign endometrial lesions differ from squamous differentiation tissue and are not infected with human papillomavirus. AB - BACKGROUND: Squamous differentiation/squamous metaplasia is often associated with endometrial adenocarcinoma and benign lesions, such as endometrial hyperplasia and chronic endometritis. Morules have distinct histological characteristics, and are referred to as squamous metaplasia or squamoid metaplasia. AIM: To focus on the histological characteristics of morules and clarify the difference between morules and squamous differentiation. MATERIALS/METHODS: Twenty endometrioid carcinomas with morules or squamous differentiation, five adenosquamous carcinomas, and eight non-carcinomatous endometrial lesions with morules were investigated. Numerous antibodies for epithelial membrane antigen (EMA), involucrin, cytokeratins, neuropeptides, and oncofetal antigens were used for immunohistochemistry. In situ hybridisation and polymerase chain reaction were used to detect human papillomavirus (HPV). RESULTS: The morules observed were uniform cell clusters, with no squamous differentiation. They were immunonegative for epithelial antigens including involucrin, EMA, and cytokeratins, but were positive for neurone specific enolase. A few morules were immunopositive for acetylcholine esterase, and one case was positive for somatostatin; neither oncofetal nor proliferative cell markers, including blood group A, B, and AB, or other neuropeptides were demonstrated in the morules. HPV DNA was not found in either the morules in the carcinomas or in the benign lesions. However, true squamous differentiation tissue in four endometrioid carcinomas and two adenosquamous carcinomas was HPV positive using in situ hybridisation. CONCLUSION: Morules are histologically distinct from squamous metaplasia/squamous differentiation tissue. Morules are thought to be neuroectodermal-like cell clusters, and are not infected with HPV. In contrast, some of the true squamous differentiation tissue was associated with HPV infection. PMID- 15333651 TI - Prognostic significance of tenascin-C expression in superficial and invasive bladder cancer. AB - AIMS: Tenascin-C (Tn-C) is an extracellular matrix glycoprotein that is upregulated in malignant tumours. Tn-C promotes cell growth, cell migration, and angiogenesis. It has been suggested to be a prognostic factor in various types of malignant tumours, but there is little information on its significance in bladder cancer with regard to overall survival (OS) and recurrence free survival (RFS). METHODS: Tn-C expression was studied in 106 patients with bladder cancer diagnosed between 1994 and 1997. Immunohistochemistry was performed using a monoclonal antibody against Tn-C. RFS and OS were estimated by the Kaplan-Meier method and compared by the log rank test in univariate analysis and by the Cox multistep regression method in multivariate analysis. RESULTS: Within the mean follow up period of 126 months, patients with diffuse Tn-C staining in the tumour stroma had a significantly worse OS than those with negative staining or only moderate Tn-C expression (p = 0.025). Patients with cytoplasmic expression of Tn C had a significantly better OS than those without (p = 0.001). Multivariate analysis, taking into consideration age, grade, stage, tumour associated carcinoma in situ, progression, and Tn-C staining in tumour stroma, showed that only expression of Tn-C in invasive tumour cells was an independent positive prognostic factor for OS (p = 0.049). CONCLUSIONS: Tn-C may provide important prognostic information in bladder cancer depending on the expression pattern in the tumour stroma or cytoplasm of the tumour cells. PMID- 15333652 TI - Expression pattern of CK7, CK20, CDX-2, and villin in intestinal-type sinonasal adenocarcinoma. AB - BACKGROUND: Intestinal-type sinonasal adenocarcinoma (ITAC) is an uncommon neoplasm, which resembles adenocarcinoma of the gastrointestinal tract. ITAC occurs sporadically or in association with occupational exposure to hardwood dust and other agents. AIMS: To investigate the phenotype and possible pathogenetic mechanisms of primary sinonasal and nasopharyngeal adenocarcinomas by staining for cytokeratin 7 (CK7), CK20, CDX-2, and villin. METHODS: Twelve sporadic sinonasal and nasopharyngeal adenocarcinomas were stained with monoclonal antibodies to CK7, CK20, CDX-2, and villin. The ITACs were classified as papillary, colonic, solid, mixed, or mucinous types. RESULTS: The diagnosis of ITAC was confirmed in 10 cases: five were colonic type and five were papillary. One was a sinonasal papillary low grade adenocarcinoma, and one a papillary nasopharyngeal adenocarcinoma, and these tumours were CK7 positive, but CK20, CDX 2, and villin negative. All ITACs were positive for CK20, CDX-2, and villin, and six were CK7 positive. One ITAC had a focus of intestinal metaplasia away from the invasive carcinoma. CONCLUSIONS: Sinonasal ITACs have a distinctive phenotype, with all cases expressing CK20, CDX-2, and villin. Most ITACs also express CK7, although a proportion of tumours are CK7 negative. ITAC seems to be preceded by intestinal metaplasia of the respiratory mucosa, which is accompanied by a switch to an intestinal phenotype. Although ITACs are morphologically similar, differences in cytokeratin expression patterns suggest two distinct types. The expression pattern of CK7, CK20, CDX-2, and villin positive may be useful in separating these tumours from other non-ITAC adenocarcinomas of the sinonasal tract and nasopharynx. PMID- 15333653 TI - Ki-ras gene mutations, LOH of the APC and DCC genes, and microsatellite instability in primary colorectal carcinoma are not associated with micrometastases in pericolonic lymph nodes or with patients' survival. AB - AIMS: The primary aim of this study was to look for possible correlations between molecular genetic changes in primary colorectal cancer and the presence or absence of micrometastases in the accompanying pericolonic lymph nodes. The secondary aim was to correlate the data on these molecular genetic changes and micrometastases with survival. METHODS: One hundred and twenty five Dukes's stage B colorectal cancers from 1989 to 1992 were analysed. The primary tumours were evaluated for Ki-ras mutation, adenomatous polyposis coli (APC) loss of heterozygosity (LOH), deleted in colon cancer (DCC) LOH, and microsatellite instability using standard molecular techniques. All available lymph nodes were immunohistochemically stained for micrometastases. RESULTS: Micrometastases were present in 41% of patients. There were significantly more lymph nodes removed in the patients with micrometastases. Micrometastases were not associated with Ki ras mutation, APC LOH, DCC LOH, or microsatellite instability, even when controlling for the number of lymph nodes removed. None of the molecular variables considered had a significant impact on either overall survival or on death with disease. CONCLUSIONS: There are insufficient data to justify using molecular genetic changes in primary colorectal carcinomas as prognostic markers. Micrometastases do not provide prognostic information on survival. There is value in increasing the numbers of lymph nodes removed and analysed along with the primary tumour. PMID- 15333654 TI - What happens to patients with positive tissue transglutaminase and endomysium antibody results in general practice? AB - BACKGROUND: A previous study showed that many patients with positive gut related antibodies (anti-tissue transglutaminase (TTG) and/or anti-endomysium (EMA) antibodies), indicative of coeliac disease, were not offered a duodenal biopsy, despite the recommendation of the British Society for Gastroenterology guidelines. AIMS/METHODS: To investigate whether the addition of a comment on the advisability of referral to a gastroenterologist and biopsy to each positive gut related antibody result would improve the referral rate to investigate possible coeliac disease. RESULTS: The referral rate improved from 30.1% of patients who were referred and 18% subsequently biopsied before the addition of the comment, to 79.8% who were referred after the introduction of a specific laboratory comment attached to positive antibodies to TTG and/or EMA. All patients with these positive antibodies who were referred for consultant opinion were subsequently biopsied. CONCLUSIONS: To ensure that British Society for Gastroenterology guidelines are followed, laboratories should incorporate more explicit details on the recommended course of action for general practitioners on their receipt of positive gut antibodies to TTG/EMA. PMID- 15333655 TI - Serrated adenomas of the appendix. AB - AIMS: A review of the literature indicated that only one case of serrated adenoma of the appendix has been recorded. The aim was to explore the possible occurrence of serrated adenomas of the appendix at the department of pathology, Karolinska Institute and University Hospital, Stockholm, Sweden. METHODS: Between January 1993 and December 2003, 38 non-carcinoid, non-neoplastic, or neoplastic polyps or tumours of the appendix were surgically removed at this hospital. All filed histological sections (haematoxylin and eosin stained) were reviewed. RESULTS: Of the 38 lesions, four were hyperplastic polyps, 10 serrated adenomas, six villous adenomas, and the remaining eight mucinous adenocarcinomas without a remnant adenoma. Serrated adenomas accounted for six of the 11 adenomas without invasion, and four of the 15 adenomas with invasive carcinoma. At the time of surgical resection, four of the 10 serrated adenomas had evolved into invasive carcinomas, in addition to 11 of the 16 villous adenomas. CONCLUSIONS: Serrated and villous adenomas of the appendix appear to be highly aggressive lesions, more aggressive than similar adenomas in the colon and rectum. Of the seven cases with a hyperplastic polyp, one concurred with a serrated adenoma, two with a serrated adenoma having an invasive carcinoma, and one with invasive carcinoma without a remnant adenomatous structure. At present, there is an increased awareness that some hyperplastic polyps of the colon and rectum may evolve into serrated adenomas. Whether this pathway is also valid for the appendix vermiformis should be investigated in a larger number of cases. PMID- 15333656 TI - Relation between lymphopenia and bacteraemia in UK adults with medical emergencies. AB - AIMS: To determine the relevance of lymphopenia to the diagnosis of bacteraemia in patients admitted with medical emergencies, relative to peripheral blood white cell count and neutrophilia. PATIENTS/METHODS: A two year cohort study carried out in a teaching hospital in Oxford, UK of 21,495 consecutive adult emergency admissions to general medical or infectious disease wards. Full blood data were available in 21,372 cases; 41 cases with extreme full blood count results (neutrophil count, > 75 x 10(9)/litre; lymphocyte count, > 10 x 10(9)/litre) were excluded, leaving 21,331 cases for analysis. The association between the admission lymphocyte and neutrophil counts and the risk of bacteraemia was assessed. RESULTS: Neutrophilia and lymphopenia were both associated with bacteraemia. Lymphopenia was the better predictor in this cohort. Both neutrophilia and lymphopenia were more predictive of bacteraemia than the total white blood cell count. CONCLUSIONS: Both lymphocyte and neutrophil counts, rather than total white blood cell count, should be considered in adult medical admissions with suspected bacteraemia. PMID- 15333657 TI - The predictive value of total serum IgE for a positive allergen specific IgE result. AB - BACKGROUND: Measurement of total serum IgE and allergen specific IgE is often requested to assess possible allergy. As public awareness increases, so do requests for allergy assessment; unless there is a clear "allergen suspect" in the history, several allergen specific IgE requests may be made. This increases the likelihood of detecting borderline increases in allergen specific IgE of uncertain relevance, and has important cost implications for the service. AIMS: To provide an evidence base for this observation. METHODS: In this retrospective observational study, results from 301 patients under 16 years of age from whom blood was taken for "allergy testing" from March 2001 to February 2003 were studied. RESULTS: Allergen specific IgE testing in children with low total serum IgE concentrations (<10 IU/litre) yielded very few positive results (three of 73 children), except in those being investigated for an acute reaction to a single food; when IgE was 11-20 kU/litre, 13 of 73 children had positive allergen specific IgE; in the 21-40 kU/litre IgE group, 16 of 74 children had positive allergen specific IgE and in the 41-80 kU/litre group, 22 of 81 had positive allergen specific IgE. CONCLUSIONS: Allergen specific IgE testing in children with low IgE concentrations (<10 kU/litre) produces few positive results in patients with non-specific symptoms. Laboratories should perform allergy testing for specific allergens regardless of total IgE concentration only when there are convincing clinical reasons to do so, and should not proceed with this if the total IgE is <10 kU/litre and the presenting symptoms are non-specific. PMID- 15333658 TI - Supervised automated microscopy increases sensitivity and efficiency of detection of sentinel node micrometastases in patients with breast cancer. AB - AIMS: To investigate the practicality and sensitivity of supervised automated microscopy (AM) for the detection of micrometastasis in sentinel lymph nodes (SLNs) from patients with breast carcinoma. METHODS: In total, 440 SLN slides (immunohistochemically stained for cytokeratin) from 86 patients were obtained from two hospitals. Samples were selected on the basis of: (1) a pathology report mentioning micrometastases or isolated tumour cells (ITCs) and (2) reported as negative nodes (N0). RESULTS: From a test set of 29 slides (12 SLN positive patients, including positive and negative nodes), 18 slides were scored positive by supervised AM and 11 were negative. Routine examination revealed 17 positive slides and 12 negative. Subsequently, automated reanalysis of 187 slides (34 patients; institute I) and 216 slides (40 patients; institute II) from reported node negative (N0) patients showed that two and seven slides (from two and five patients, respectively) contained ITCs, respectively, all confirmed by the pathologists, corresponding to 5.9% and 12.5% missed patients. In four of the seven missed cases from institute II, AM also detected clusters of four to 30 cells, but all with a size < or = 0.2 mm. CONCLUSIONS: Supervised AM is a more sensitive method for detecting immunohistochemically stained micrometastasis and ITCs in SLNs than routine pathology. However, the clinical relevance of detecting cytokeratin positive cells in SLNs of patients with breast cancer is still an unresolved issue and is at the moment being validated in larger clinical trials. PMID- 15333659 TI - CD133 positive endothelial progenitor cells contribute to the tumour vasculature in non-small cell lung cancer. AB - AIMS: Recent results generated in a mouse model suggest that tumour angiogenesis/vasculogenesis can be initiated and maintained by bone marrow derived endothelial progenitor cells. This present study investigated the distribution and frequency of CD133 positive endothelial progenitor cells in patients with non-small cell lung cancer (NSCLC) (tumour tissue and tumour free lung regions) and healthy controls using fresh frozen specimens. The novel marker CD133 identifies human haemopoetic precursor cells, in addition to human endothelial progenitor cells. METHODS: Seventy nine lung cancer specimens and 66 adjacent histologically tumour free tissues of the same patient cohort were analysed; 11 postmortem specimens from control patients who did not suffer from malignant disease served as controls. Cryostat sections were stained for CD133, CD31, vascular endothelial growth factor receptor 2 (VEGFR-2; KDR), p53, and the proliferation marker Ki-67, and the correlations were analysed. RESULTS: Forty three of 63 evaluable tumour specimens had increased numbers of CD133 positive cells and in some cases capillary forming CD133 positive structures were detectable. In addition, 30 of 63 specimens had raised expression of KDR and 29 of 63 had increased MVD. Increased CD133 expression marginally correlated with raised KDR expression but not with p53 and Ki-67. CONCLUSION: A significant increase in CD133 positive cells was documented in patients with NSCLC, suggesting an involvement of endothelial progenitor cells in tumour vasculogenesis and tumour growth in these patients. PMID- 15333660 TI - Presence of vascular adventitial fibroblastic cells in diffuse-type gastric carcinomas. AB - AIM: To investigate morphological changes in the tumour vessel adventitia, particularly the distribution of vascular adventitial fibroblastic cells (VAFCs)- namely, CD34 positive fibroblastic cells just outside the vascular media--in diffuse-type gastric carcinomas. METHOD: In total, 18 surgically resected advanced typical diffuse-type gastric carcinomas and their normal tissues were examined. Immunostaining for CD34, CD31, high molecular weight caldesmon (HCD), and cytokeratin 8 (CAM5.2) was performed to detect VAFCs. VAFCs are positive for CD34 but negative for CD31, and are located just outside the vascular media (HCD positive vascular smooth muscle bundle). The areas just outside the vascular media in the whole maximum tumour cut surface were assessed, except the tumour growing edge, which was confirmed by immunostaining with CAM5.2. CD34 positive and CD31 negative cells just outside the vascular media were defined as VAFCs. RESULTS: VAFC containing vessels were seen in 17 of the 18 diffuse carcinoma tissues. Vessels lacking VAFCs were also detected in these 17 tumours. In contrast, all of the vessels lacked VAFCs in the remaining tumour. In the 18 samples of normal tissue, all of the vessels contained VAFCs. CONCLUSIONS: These results suggest that the presence of VAFCs is associated with the infiltration of diffuse scattered gastric carcinoma cells. PMID- 15333661 TI - Colonic IgA producing cells and macrophages are reduced in recurrent and non recurrent Clostridium difficile associated diarrhoea. AB - BACKGROUND: In Clostridium difficile associated diarrhoea (CDAD), histological changes in the colonic mucosa range from minimal inflammation to pseudomembranous colitis (PMC). The disease also recurs in a considerable proportion of patients. AIM: To investigate mucosal immune system cells in colonic biopsies of patients with CDAD. METHODS: Colonic biopsies were obtained from 12 control patients with diarrhoea, six patients with CDAD and minimal inflammation, and 10 patients with CDAD with pseudomembranous colitis (samples obtained from areas with and without inflammatory exudate). Immunohistochemical studies were performed using antibodies to T cells (CD3), macrophages (CD68), B/plasma cells (CD79alpha), and to IgA, IgM, and IgG. Labelled cells in lamina propria were quantified. RESULTS: In contrast to T cells, there were significant reductions in B/plasma cell and macrophage counts in all biopsies from patients with CDAD compared with controls (p<0.001). Studies using anti-immunoglobulin antibodies showed significant reductions in IgA producing cells in CDAD biopsies (p<0.05), with the greatest reduction in samples from patients with PMC. In contrast, there was a significant increase (p<0.05) in IgG producing cells in CDAD biopsies. Only patients with PMC relapsed. In these patients, B/plasma cell and IgA producing cell counts (in biopsies with and without inflammatory exudates) were significantly lower (p<0.01) in mucosal samples from those who subsequently relapsed (five) than those who did not. CONCLUSIONS: A selective reduction in mucosal IgA producing cells and macrophages is associated with colonic disease in C difficile infected patients. Severe reduction in colonic IgA producing cells may predispose to recurrence of CDAD. PMID- 15333662 TI - Discrepancies between clinical and postmortem diagnoses in Jamaica: a study from the University Hospital of the West Indies. AB - AIMS: It has previously been shown that the low necropsy request rate at the University Hospital of the West Indies (UHWI) in Jamaica (35.3%) results primarily from clinicians' confidence in clinical diagnoses and laboratory investigations. This study aimed to determine the rates of discrepancy between clinical and necropsy diagnoses at the UHWI, because many previous studies from other institutions have shown persistent high rates of discrepancy, despite advances in medical investigative technology over the past several years. METHODS: Data were extracted retrospectively from consecutive necropsies performed at the UHWI over a two year period. The data were analysed to determine the categories and rates of discrepancy, and to determine the relation between discrepancy rates and age, sex, type and number of diagnoses for each patient, hospital service, and length of hospitalisation. RESULTS: Necropsies were performed on 446 patients; 348 were suitable for further analysis. The overall discrepancy rate was 48.4% and the diagnoses with the highest individual discrepancy rates were pneumonia (73.5%), pulmonary thromboembolism (68.3%), and myocardial infarction (66.7%). Males and older patients were more likely to have discrepant diagnoses. There was a high frequency of discrepancies in patients who died within 24 hours of admission, but there was no consistent relation between length of hospitalisation and discrepancy rate. CONCLUSIONS: The high discrepancy rates documented at the UHWI are similar to those reported globally. This study supports previous attestations that the necropsy remains a vital tool for determining diagnostic accuracy, despite modern modalities of clinical investigation and diagnosis. PMID- 15333663 TI - A primary amelanotic melanoma of the vagina, diagnosed by immunohistochemical staining with HMB-45, which recurred as a pigmented melanoma. AB - Usually, malignant melanoma is readily diagnosed by the presence of melanin granules. Although amelanotic melanoma contains a few melanin granules, it is often difficult to differentiate from non-epithelial malignant tumours. This report describes a case of amelanotic melanoma of the vagina, which was originally suspected to be a non-epithelial malignant tumour, but was subsequently correctly diagnosed by immunohistochemical staining with the HMB-45 antibody and for the S-100 protein. A light grey tumour with superficial ulceration was located in the upper third of the vagina. The patient was treated with irradiation followed by chemotherapy. Subsequently, the tumour disappeared and cytology was negative; thus, she achieved complete remission. However, 20 months after complete remission, the tumour recurred locally: the site had a grossly black appearance, which was pathognomonic for a malignant melanoma. Thus, HMB-45 and S-100 protein immunohistochemistry confirmed the diagnosis of amelanotic melanoma. PMID- 15333664 TI - Statin precipitated lactic acidosis? AB - An 82 year old woman was admitted with worsening dyspnoea. Arterial blood gases were taken on air and revealed a pH of 7.39, with a partial pressure of CO2 (pCO2) of 1.2 kPa, pO2 of 19.3 kPa, HCO3 of 13.8 mmol/litre, and base excess of 16.3 mmol/litre: a compensated metabolic acidosis with hyperventilation induced hypocapnia, which is known to be a feature of lactic acidosis. There was also an increased anion gap ((Na140 + K4.0) - (Cl 106 + HCO3 13.8) = 24.2 mEq/litre (reference range, 7-16)), consistent with unmeasured cation. Lactate was measured and found to be raised at 3.33 mmol/litre (reference range, 0.9-1.7). After exclusion of common causes of lactic acidosis Atorvastatin was stopped and her acid-base balance returned to normal. Subsequently, thiamine was also shown to be deficient. The acidosis was thought to have been the result of a mitochondrial defect caused by a deficiency of two cofactors, namely: ubiquinone (as a result of inhibition by statin) and thiamine (as a result of dietary deficiency). PMID- 15333665 TI - A fatal case of disseminated aspergillosis caused by a non-sporulating strain of Aspergillus fumigatus. AB - This report describes the case of a 38 year old pregnant woman with fatal disseminated aspergillosis and multiorgan failure, which was preceded by a long history of allergic bronchopulmonary aspergillosis. Postmortem revealed massive infarction and abscess formation in both lungs. Histology revealed a focal granulomatous response. Fungal infiltration with areas of necrosis were also seen in the liver, spleen, and paratracheal, mediastinal, para-aortic, and hilar lymph nodes. Culture of tissue samples produced a non-sporulating, beige coloured fungus that developed green pigmentation only after three weeks of incubation. Nucleotide sequencing of the D1-D2 region of the large ribosomal subunit revealed 100% homology with Aspergillus fumigatus. Minimum inhibitory concentrations for amphotericin B and itraconazole were both 0.25 mg/litre (susceptible). Further work is urgently required to determine the prevalence of such non-sporulating strains and their relevance to clinical infection. PMID- 15333666 TI - Mediastinal hibernoma: a case report. AB - Hibernomas are rare benign tumours that arise most often in adults from the remnants of fetal brown adipose tissue. They usually affect muscle and subcutaneous tissue and are asymptomatic and slow growing. The distribution of this tumour follows the sites of persistence of brown fat. Out of more then 100 cases described in the word literature only three hybernomas were mediastinal. A recent clinicopathological study of 170 cases from the Armed Forces Institute of Pathology confirmed the exceptionality of the intrathoracic location. This report describes a very rare case of mediastinal hibernoma in a young man. PMID- 15333667 TI - Resolution of chronic severe refractory thrombocytopenia after treatment of hypothyroidism. AB - The case of a 52 year old woman with chronic severe refractory thrombocytopenia is presented. Over a three year period, her platelet count was persistently less than 20 x 10(9)/litre (normal range, 150-400). She required repeated hospital admission for management of bleeding and received multiple blood transfusions. She was given repeated courses of steroids, immunosuppression, immunoglobulin, and splenectomy, without success, in an attempt to stop the chronic blood loss. Eventually, she was found to be profoundly hypothyroid. On correction of her thyroid deficiency the platelet count returned to the normal range and all bleeding stopped. The platelet count remains in the normal range three years later. PMID- 15333668 TI - Ciliary body adenoma of non-pigmented epithelium. AB - Adenomas of the non-pigmented epithelium of the ciliary body are rare neoplasms and most of the studies are in the form of case reports. There are only 27 documented cases of acquired neoplasm of the non-pigmented ciliary epithelium (NPCE) reported in the English literature. In most reports, there was a clinical suspicion of melanoma and the diagnosis of NPCE adenoma was made on histopathological evaluation of the resected tissue. The entity has not been reported in the Pakistani population to date. This report describes a case of ciliary body adenoma of NPCE in a 27 year old Pakistani man. The histological and immunohistochemical profiles were typical of the adenomas described in the literature. PMID- 15333669 TI - Myelofibrosis and angiodysplasia of the colon: another manifestation of portal hypertension and massive splenomegaly? AB - Bleeding owing to portal hypertensive colopathy, a form of large bowel angiodysplasia, as a cause of increased blood transfusion requirement is described in a 74 year old man with idiopathic myelofibrosis. The proposed mechanism and the potential therapeutic options for this rare complication of myelofibrosis are discussed. PMID- 15333670 TI - Pathology of astrovirus associated diarrhoea in a paediatric bone marrow transplant recipient. AB - Human astrovirus infection often causes outbreaks of self limiting diarrhoea, but may also infect patients who are immunodeficient or immunocompromised. Although there are previous publications relating to various aspects of astroviruses, there is a minimal amount of literature on the histopathological features of gastrointestinal astrovirus infection in humans. We report the histopathological findings, including immunohistochemical and electron microscopic features, of astrovirus infection in a bone marrow transplant recipient aged 4 years with diarrhoea. The appearance of a small intestinal biopsy did not suggest graft versus host disease, but demonstrated villous blunting, irregularity of surface epithelial cells, and an increase in lamina propria inflammatory cell density. Immunohistochemical staining with a murine astrovirus group specific monoclonal antibody demonstrated progressively more extensive staining in the duodenal and jejunal biopsies, predominantly restricted to the luminal surface and cytoplasm of surface epithelial cells, most marked at the villus tips. Electron microscopic examination demonstrated viral particles within the cytoplasm of enterocytes, focally forming paracrystalline arrays. PMID- 15333671 TI - A case of 49,XXXXX in which the extra X chromosomes were maternal in origin. AB - This report describes an 11 month old female baby with features of pentasomy X. A molecular and cytogenetic evaluation revealed that her karyotype was 49,XXXXX and her extra X chromosomes were of maternal origin. She has muscular hypotonia, mental retardation, a cleft palate, mild hydrocephalus as a result of dilatation of both lateral ventricles, hyperextensible elbow joints, proximal radioulnar synostosis, clinodactyly of the fifth finger, valgus of the feet, and small hands and feet. In addition, she has a persistent pupillary membrane and congenital chorioretinal atrophy. The pathogenesis of pentasomy X is not clear at present, but it is thought to be caused by successive maternal non-dysjunctions. PMID- 15333672 TI - Lymphadenoma of the salivary gland: a rare tumour. PMID- 15333673 TI - Gastric precancerous lesion follow up based on pathological evidence. PMID- 15333674 TI - Quantitative assessment of a novel flow-through porous microarray for the rapid analysis of gene expression profiles. AB - A novel microarray system that utilizes a porous aluminum-oxide substrate and flow-through incubation has been developed for rapid molecular biological testing. To assess its utility in gene expression analysis, we determined hybridization kinetics, variability, sensitivity and dynamic range of the system using amplified RNA. To show the feasibility with complex biological RNA, we subjected Jurkat cells to heat-shock treatment and analyzed the transcriptional regulation of 23 genes. We found that trends (regulation or no change) acquired on this platform are in good agreement with data obtained from real-time quantitative PCR and Affymetrix GeneChips. Additionally, the system demonstrates a linear dynamic range of 3 orders of magnitude and at least 10-fold decreased hybridization time compared to conventional microarrays. The minimum amount of transcript that could be detected in 20 microl volume is 2-5 amol, which enables the detection of 1 in 300,000 copies of a transcript in 1 microg of amplified RNA. Hybridization and subsequent analysis are completed within 2 h. Replicate hybridizations on 24 identical arrays with two complex biological samples revealed a mean coefficient of variation of 11.6%. This study shows the potential of flow-through porous microarrays for the rapid analysis of gene expression profiles in clinical applications. PMID- 15333675 TI - Comprehensive comparison of six microarray technologies. AB - Microarray technology is extensively used in biological research. The applied technologies vary greatly between laboratories, and outstanding questions remain regarding the degree of correlation among approaches. Recently, there has been a drive toward ensuring high-quality microarray data by the implementation of MIAME (Minimal Information About a Microarray Experiment) guidelines and an emphasis on ensuring public-availability to all datasets. However, despite its current widespread use and availability, very little is known about the extent to which application of the different technologies influences the outcome of transcriptional profiles and differential expression. The results among the handful of published studies are conflicting. Here, we present a comprehensive evaluation encompassing different reporter systems (short oligonucleotides, long oligonucleotides and cDNAs), labelling techniques and hybridization protocols. We used four oligonucleotide and two cDNA platforms to compare gene expression between two sample types. We determined the overall consistency (reproducibility) within each platform, and correlation among replicates within and between technologies. We find that the top performing platforms show low levels of technical variability that result in an increased ability to detect differential expression. Most importantly, we show the top four platforms are highly correlated with biological, rather than technological, differences accounting for the majority of variation in the data. PMID- 15333676 TI - Opioid receptor involvement in food deprivation-induced feeding: evaluation of selective antagonist and antisense oligodeoxynucleotide probe effects in mice and rats. AB - Central administration of general and selective opioid receptor subtype antagonists in the rat has revealed a substantial role for mu, a moderate role for kappa, and a minimal role for delta receptors in the mediation of deprivation induced feeding. Antisense probes directed against the kappa opioid receptor (KOP), nociceptin opioid receptor (NOP), and delta opioid receptor (DOP) genes in rats result in reductions similar to kappa and delta antagonists, whereas antisense probes directed against the mu opioid receptor (MOP) gene produced modest reductions relative to mu antagonists, suggesting that isoforms of the MOP gene may mediate deprivation-induced feeding. Since these isoforms were initially identified in mice, the present study compared the effects of general and selective opioid receptor antagonists on deprivation-induced feeding in rats and mice and antisense probes directed against exons of the MOP, DOP, KOP, and NOP genes on deprivation-induced feeding in the mouse. Food-deprived (12 and 24 h) rats and mice displayed similar profiles of reductions in deprivation-induced feeding following general, mu, and kappa opioid antagonists. In contrast, mice, but not rats, displayed reductions in deprivation-induced intake following delta antagonism as well as DOP antisense probes, suggesting a species-specific role for the delta receptor. Antisense probes directed against the KOP and NOP genes also reduced deprivation-induced intake in mice in a manner similar to kappa antagonism. However, the significant reductions in deprivation-induced feeding following antisense probes directed against either exons 2, 4, 7, 8, or 13 of the MOP gene were modest compared with mu antagonism, suggesting a role for multiple mu-mediated mechanisms. PMID- 15333677 TI - GABAB receptor antagonist-mediated antidepressant-like behavior is serotonin dependent. AB - There is an emerging body of data purporting a role of gamma-aminobutyric acid (GABA) in the pathophysiology of mood disorders. However, the role of metabotropic GABA(B) receptors in depression is not well defined. The modified forced swim test has recently emerged as an excellent tool to assess behaviorally the role of monoamines in antidepressant action. To assess the role of GABA(B) receptors in antidepressant-related behavior, we examined a number of selective GABA(B) receptor ligands (novel positive modulators and antagonists) on behavior in the modified forced swim test. We demonstrate that the selective GABA(B) receptor antagonists CGP56433A [[3-{1-(S)-[{3-cyclohexylmethyl)hydroxy phosphinyl}-2-(S) hydroxy propyl]amino}ethyl]benzoic acid; 1-10 mg/kg] and [3-[[1 (S)-3-dichlorophenyl)ethyl]amino]-2-(S)-hydroxy-propyl]phenylmethyl-phosphinic acid hydrochloride; 3-10 mg/kg] had a similar profile to the selective serotonin reuptake inhibitor fluoxetine; they decreased immobility and increased swimming behavior. The tricyclic antidepressant desipramine decreased immobility but increased climbing behavior. In contrast, the novel GABA(B) receptor-positive modulator GS39783 (10-40 mg/kg) did not display antidepressant-like activity in the modified forced swim test. To further assess the possible interaction between GABA(B) receptor antagonism and serotonin, rats were pretreated with the tryptophan hydroxylase inhibitor para-chlorophenylalanine. 5-Hydroxytryptamine depletion (>90%) abolished the antidepressant-like behavior of CGP56433A (10 mg/kg) by attenuating the increase in swimming. Together, these data demonstrate that GABA(B) receptor antagonists via an interaction with the serotonergic system display antidepressant-like properties and therefore represent a novel approach for the treatment of depression. PMID- 15333678 TI - Regulation of M2 muscarinic acetylcholine receptor expression and signaling by prolonged exposure to allosteric modulators. AB - The effects of prolonged exposure of M(2) muscarinic acetylcholine receptors (mAChRs), stably expressed in Chinese hamster ovary cells, to the allosteric modulators gallamine, alcuronium, and heptane-1,7-bis (dimethyl-3' phthalimidopropyl)-ammonium bromide (C(7)/3'-phth) were compared with the effects of the agonist carbachol (CCh) and antagonists atropine and N-methylscopolamine (NMS). Intact cell saturation binding assays using [(3)H]NMS found that pretreatment of the cells for 24 h with CCh caused a significant down-regulation of receptor number, whereas atropine, NMS, and all three allosteric modulators caused receptor up-regulation. Functional assays using a cytosensor microphysiometer to measure whole-cell metabolic rate found no acute effects of gallamine on receptor signaling, whereas atropine seemed to behave as an inverse agonist. Pretreatment of the cells with gallamine (20 microM) or atropine (20 nM) resulted in a significant enhancement of the maximal effect evoked by CCh. In contrast, CCh (100 microM) pretreatment resulted in a significant reduction in maximal receptor signaling capacity. Time-course experiments revealed that the effects of atropine and gallamine on receptor up-regulation are only visualized after at least 12-h ligand exposure, compared with the more rapid effects of CCh, which achieve steady-state down-regulation within 90 min. Additional experiments monitoring CCh-mediated M(2) mAChR internalization in the presence of gallamine revealed that part of the mechanism underlying the effects of the modulator on receptor expression may involve a change in receptor internalization properties. These findings suggest that, like orthosteric ligands, G protein-coupled receptor allosteric modulators also are able to mediate long-term effects on receptor regulation. PMID- 15333679 TI - Superagonism at the human somatostatin receptor subtype 4. AB - We have discovered a novel compound, J-2156 [(1'S, 2S)-4-amino-N-(1'-carbamoyl-2' phenylethyl)-2-(4''-methyl-1''-naphthalenesulfonylamino)butanamide], that belongs to a new class of somatostatin receptor ligands. J-2156 binds with nanomolar affinity to the human somatostatin receptor subtype 4 and is over 400-fold subtype-selective against the other somatostatin receptors. When evaluated in a [(35)S]guanosine-5'-O-(3-thio) triphosphate binding assay, J-2156 elicited a response 2 to 3 times as large as that of somatostatin-28 and somatostatin-14. That somatostatin-14 is clearly not a maximally efficacious agonist could be verified by demonstrating that it displays the typical behavior of a partial agonist when tested against J-2156. Increasing concentrations of somatostatin-14 cause a concentration-dependent rightward shift of the dose-response curves for J 2156, without affecting its maximal response. This lack of reduction of the maximal response and the fact that the superior efficacy of J-2156 is detected in membranes argue against desensitization and internalization as possible explanations for the superior efficacy of J-2156. More likely is that somatostatin-14 and J-2156 stabilize distinct receptor conformations that differ in their ability to interact with G-proteins. In a cyclic AMP assay, J-2156, somatostatin-28, and somatostatin-14 all act as full agonists. However, this outcome is most likely due to the presence of a receptor reserve in the cyclic AMP assay since there is a large gain of apparent potency in the cyclic AMP assay and the gain is larger for J-2156 than for somatostatin. We conclude that the endogenous ligands somatostatin-14 and somatostatin-28 do not define maximal agonism on the human somatostatin receptor subtype 4 and that J-2156 represents a so-called superagonist. PMID- 15333680 TI - Treatment of invasive bladder cancer: lessons from the past and perspective for the future. AB - Radical cystectomy with lymphadenectomy is the gold standard for treatment of invasive bladder cancer. However, the treatment alone does not always provide a satisfactory result for the disease extending outside the bladder. In this review we discuss several clinical issues in the diagnosis and treatment of this invasive disease. Although the quality of diagnostic imaging modalities has improved, they are still not sensitive enough for the staging of the disease, especially for early invasive disease. In addition, lack of serum markers hinders appropriate monitoring of patients with the disease. Regarding the surgical aspect of lymphadenectomy, the area of its dissection, the standard number of nodes retrieved and the method of pathological examination should be established so that the clinical benefits of surgery can be more clearly defined. Neoadjuvant chemotherapy for invasive disease is promising for improvement of survival of patients. A chemotherapy regimen as effective as, but less toxic than, MVAC (methotrexate, vinblastine, doxorubicin and cisplatin) has been reported and several phase III clinical trials have been launched to determine the benefits of adjuvant or neoadjuvant chemotherapy with newly developed agents. However, we still lack a chemotherapy regimen more effective than MVAC, which is the most crucial issue in the treatment of this invasive disease. An alternative option for such disease may be bladder preservation with transurethral resection of tumor followed by chemoradiotherapy. However, patients who are indicated for this treatment may be limited to those with early invasive disease having certain favorable clinical and pathological features. PMID- 15333681 TI - Serum EBV DNA as a biomarker in primary nasopharyngeal carcinoma of Indian origin. AB - BACKGROUND: Nasopharyngeal carcinoma (NPC) is a unique tumor due to its etiology and endemic distribution. Ethnic and regional factors are found to strongly influence the risk of disease; however, there have been no well-conducted studies on Indian patients. The present study assesses the relationship between Epstein Barr Virus (EBV) and sporadic Indian NPC and the role of serum EBV DNA in NPC detection. METHODS: Primers directed against non-polymorphic Epstein-Barr nuclear antigen-1 (EBNA-1) gene were used to detect the presence of EBV DNA from fresh tissue and serum in NPC, using PCR. RESULTS: EBV DNA was detected in 69% of the biopsies and 58% of the serum of the NPC patients. With respect to histology, WHO Type III NPC, WHO Type II tumors and WHO I tumors showed 100%, 72.2% and 33% EBV positivity, respectively. EBV positivity was also observed in 23% (6/26) of benign samples. All biopsies of patients with positive serum samples were positive for EBV DNA. CONCLUSION: EBV infection was found in sporadic NPC of South Indian origin, which confirms the etiological role of EBV in NPC. Detection of EBNA-1 in the serum and corresponding tissues of NPC patients suggests that the serum EBV DNA originates from NPC and also indicates the benefit of circulating viral DNA as an early marker in the diagnosis of NPC. Serum DNA-PCR methods can be extrapolated to follow-up studies involving tumor regression or to assess the response to various therapies. PMID- 15333682 TI - Hyperfractionated radiation therapy for oropharyngeal carcinoma in a Japanese population. AB - BACKGROUND: Hyperfractionated radiation therapy is recognized to be a better treatment method, especially regarding local control, than conventional radiation therapy for oropharyngeal carcinoma. However, the survival benefit of hyperfractionated radiation therapy for oropharyngeal carcinoma has not been reported so far. Moreover, patient populations of previous studies were mostly white or African-American, and rarely included Orientals. Thus, the current study is designed to evaluate not only local control but the survival benefit of hyperfractionated radiation therapy for oropharyngeal carcinoma in a Japanese population. METHODS: The eligibility criteria were as follows. The patients were 20 years or older. Performance status ranged from 0 to 2. No patient with double carcinoma was included except those with superficial esophageal carcinoma or advanced carcinoma controlled for at least 2 years after treatment. In our radiation protocol, hyperfractionated radiation therapy was adopted using 1.2 Gy per fraction, two fractions per day (6 h apart), 5 days a week, for a total dose of 66 Gy or more. RESULTS: Complete response was achieved in all 14 patients with oropharyngeal lesions. In five patients with lymph node metastasis, complete response was achieved in four patients and partial response was achieved in the remaining one. The 3-year loco-regional control rate was 77% and the 3-year overall survival rate was 69%. CONCLUSIONS: Treatment with hyperfractionated radiation therapy is considered superior to conventional radiation therapy with acceptable toxicity for Japanese patients with oropharyngeal carcinoma in terms of loco-regional control rate and overall survival rate. PMID- 15333683 TI - Phase II study of sequential methotrexate and 5-fluorouracil chemotherapy against peritoneally disseminated gastric cancer with malignant ascites: a report from the Gastrointestinal Oncology Study Group of the Japan Clinical Oncology Group, JCOG 9603 Trial. AB - BACKGROUND: The efficacy of systemic chemotherapy against peritoneal dissemination from advanced gastric cancer (AGC) remains unclear, because the peritoneal dissemination was not defined as a measurable lesion in conventional phase II studies. In this study, we evaluated the efficacy and toxicity of sequential MTX and 5FU therapy (MF) in chemotherapy-naive patients with AGC accompanied by malignant ascites in a phase II setting. METHODS: The treatment schedule comprised weekly administration of MTX (100 mg/m2, i.v. bolus) followed by 5FU (600 mg/m2, i.v. bolus) with a 3 h interval. Leucovorin rescue (10 mg/m2 every 6 h, for a total of six times) was commenced 24 h after MTX administration. RESULTS: Thirty-seven chemotherapy-naive patients with AGC presenting with malignant ascites were enrolled in this trial. The median age was 60 years (range, 25-74 years) and most patients (86%) had a performance status of 0-1. In total, 355 administrations of the sequential MTX/5FU therapy were performed. Major toxicity consisted of myelosuppression and gastrointestinal toxicity. Grade 4 neutropenia occurred in 10.8% of the patients. The overall objective response rate was 5.7% (two partial responses in 35 patients; 95% confidence interval: 0.7 19.2%). However, the response rate of ascites was 35.1% (complete disappearance in three patients and apparent decrease in 10 patients; 95% confidence interval: 20.2-52.5%). CONCLUSIONS: Sequential MTX/5FU therapy is effective against AGC with malignant ascites with acceptable toxicity and warrants further investigations in a phase III setting. PMID- 15333684 TI - Identification of patients with high-risk lymph node-negative colorectal cancer and potential benefit from adjuvant chemotherapy. AB - BACKGROUND: Adjuvant chemotherapy is not indicated in lymph node-negative colorectal adenocarcinoma (CRC), even though some cases will present recurrent disease. It is important to identify a subgroup of patients with the highest risk of relapse because of the potential benefit of adjuvant chemotherapy. The objective of this study is to define the prognostic factors and describe a method for the selection of this subgroup. METHODS: A retrospective cohort of 124 patients with lymph node-negative CRC with complete surgical resection was studied. Cox's proportional hazards model was used to define the prognostic factors associated with CRC-related survival and to develop a method for prediction of recurrence probability. RESULTS: The cohort included 62 women and 62 men with mean age 55.8 years. The mean follow-up period was 11.7 years. T classification of the primary tumor, differentiation grade, carcinoembryonic antigen level, gender and the presence of neural invasion were significant prognostic factors according to the multivariate analysis (final model P=0.00001). Using risk ratios for these prognostic factors, we defined a high risk group of 78 patients and a low-risk group of 46 patients with 24 and 5 recurrences, respectively (recurrence rates of 30.8% and 10.9% respectively, P=0.011). CONCLUSIONS: Using these factors, a prognostic scale was developed to predict high risk of recurrence in cases of completely resected CRC and to identify them as a subgroup of patients with potential benefit of adjuvant chemotherapy. PMID- 15333685 TI - Characteristics of patients with prostate cancer who have initially been treated by hormone therapy in Japan: J-CaP surveillance. AB - OBJECTIVE: Hormone therapy for prostate cancer has empirically prevailed in Japan. We planned to evaluate the trends and outcome of hormone therapy for establishing an adequate guideline. METHODS: Patients with prostate cancer who were initially treated by hormone therapy were registered through the J-CaP registration system. This report summarizes the background factors. RESULTS: From January 2001 to October 2003, 17,872 patients were registered from 395 institutes throughout Japan. The background factors of 17,312 patients were analyzed. The 17,872 patients were estimated as composing more than half of newly diagnosed prostate cancer patients in Japan. Of these, 22.9, 35.1, 32.9 and 8.6% belonged to T1, T2, T3 and T4, respectively. For the purposes of hormone therapy, 77.5% was primary hormone therapy. Neoadjuvant setting and adjuvant setting were 18.1 and 4.3%, respectively. About 60% of the hormone therapy was combined hormone therapy with LH-RHa plus anti-androgens. CONCLUSION: Irrespective of patients' age, TNM, stage of illness, or histological background, the majority of prostate cancer patients in Japan are receiving hormone therapy. It is necessary to evaluate whether this trend is merely a continuation of past experience of Japanese urologists or if there is a difference in the profile of effect and side effect in the case of Japanese patients compared to therapy given in Westerners. PMID- 15333686 TI - A modified low-dose regimen of mitoxantrone and prednisolone in patients with androgen-independent prostate cancer. AB - BACKGROUND: We conducted this retrospective study to analyze a modified dose schedule of mitoxantrone and prednisolone (MP) in patients with androgen independent prostate cancer. METHODS: From June 1997 to April 2002, 28 patients were enrolled. Their median age was 69 years (range, 58-79 years). The median duration of hormonal therapy was 30 months (range, 6-84 months). The median performance status was 2. Sixteen of the patients had bone disease only. The chemotherapy consisted of 8 mg/m2 mitoxantrone by intravenous infusion every 3 weeks and 10 mg prednisolone orally twice per day. WHO response criteria, prostatic-specific antigen (PSA), pain and performance status were used to assess the response. RESULTS: The median number of treatment cycles was six (range, 2 20). Nine (32.1%) and 15 patients (53.8%) had > or =80% and > or =50% reduction in serum PSA level, respectively. Of 16 patients using narcotics, five (31.3%) had a > or =50% reduction in narcotics consumption compared with the baseline. Nine patients (32.1%) showed improved performance. For 12 patients with measurable disease, only two (16.7%) showed a partial response. Grade 3-4 toxicities included neutropenia (three patients), anemia (three patients) and vomiting (one patient). The median survival was 12 months and the median time to PSA progression was 4 months. CONCLUSIONS: This modified regimen is feasible for palliative intent. The toxicity of this regimen is manageable. Exploring further combinations of this regimen with novel agents against androgen-independent prostate cancer is warranted. PMID- 15333687 TI - A randomized phase II clinical trial of tailored CPT-11 + S-1 vs S-1 in patients with advanced or recurrent gastric carcinoma as the first line chemotherapy. AB - A randomized phase II clinical trial has been designed and started in the Japanese Foundation for Multidisciplinary Treatment of Cancer (JFMC) to select the better regimen between tailored CPT-11 + S-1 and the standard S-1 treatment for advanced or recurrent gastric cancer as the first line chemotherapy. Selection of the better treatment for general clinical practice in this clinical trial will lead to a more precise assignment of a promising regimen for a future phase III randomized trial, placing continuous 5-FU infusion as the reference arm. In this trial, subsidiary pharmacokinetic analysis for the tailored dose arm is also proposed. In order to continue chemotherapy for a long time and to obtain longer survival, our study design for the tailored therapy could be exploited, especially in the field of general clinical practice. PMID- 15333688 TI - A case of postoperative recurrence of fibrolamellar hepatocellular carcinoma with increased vitamin B12 binding capacity in a young Japanese female. AB - A 17-year-old Japanese female underwent major hepatic resection for a huge fibrolamellar hepatocellular carcinoma that was compressing the inferior vena cava. The tumor was not exposed at the surgical margin but was very close to it. A recurrent lesion at the surgical margin of the liver and a lymph node metastasis were discovered 9 months postoperatively together with a marked elevation of vitamin B12 binding capacity. These lesions were resected, and vitamin B12 binding capacity decreased thereafter. Peritoneal dissemination was detected by CT 16 months postoperatively, together with recurrent elevation of vitamin B12 binding capacity. Several types of chemotherapy, including intraperitoneal injection of epirubicin, were applied and improved the patient's quality of life somewhat, but the patient died of recurrent disease 34 months after the initial hepatic resection. This is the first report in Japan of fibrolamellar hepatocellular carcinoma with increased vitamin B12 binding capacity as a useful marker. Fibrolamellar hepatocellular carcinomas, if resected, have a better prognosis than ordinary hepatocellular carcinoma in Japan, as well as in Western countries. An aggressive strategy should be chosen, which consists mainly of precise surgical resection and postoperative multimodality therapy, including chemotherapy. PMID- 15333689 TI - Cancer incidence and incidence rates in Japan in 1999: estimates based on data from 11 population-based cancer registries. PMID- 15333690 TI - Cancer statistics digest. PMID- 15333691 TI - Splicing affects presentation of RNA dimerization signals in HIV-2 in vitro. AB - During retroviral replication, full-length viral RNAs are encapsidated into new virus particles, while spliced RNAs are excluded. The Retroviridae are unique among viruses in that infectious viral particles contain a dimer of two identical genomic RNA strands. A variety of experimental data has suggested that dimerization and encapsidation of full-length viral RNAs are linked processes, although whether dimerization is a prerequisite for encapsidation, or conversely, dimerization follows encapsidation, has not been firmly established. If dimerization was the sole determinant for encapsidation, then spliced viral RNAs might be expected to display a reduced capacity for dimerization, resulting in their exclusion from the dimerization pool. Here, we studied the in vitro dimerization properties of unspliced and spliced HIV-2 RNA. We find that the rate and yield of dimerization of Nef, Rev and Tat spliced RNAs exceeded those of unspliced RNA. Although these data do not support a simple correlation between dimerization efficiency and the presence of introns, they establish that splicing affects the presentation of dimerization signal(s), which we corroborate with structure probing. This change in RNA conformation likely affects the RNA's suitability for packaging. Furthermore, the presence of upstream and downstream elements that affect the conformation of the packaging signal represents a potentially efficient viral strategy for correctly sorting spliced versus unspliced RNAs. PMID- 15333692 TI - Nucleotide-dependent interactions between a fork junction-RNA polymerase complex and an AAA+ transcriptional activator protein. AB - Enhancer-dependent transcriptional activators that act upon the sigma54 bacterial RNA polymerase holoenzyme belong to the extensive AAA+ superfamily of mechanochemical ATPases. Formation and collapse of the transition state for ATP hydrolysis engenders direct interactions between AAA+ activators and the sigma54 factor, required for RNA polymerase isomerization. A DNA fork junction structure present within closed complexes serves as a nucleation point for the DNA melting seen in open promoter complexes and restricts spontaneous activator-independent RNA polymerase isomerization. We now provide physical evidence showing that the ADP.AlF(x) bound form of the AAA+ domain of the transcriptional activator protein PspF changes interactions between sigma54-RNA polymerase and a DNA fork junction structure present in the closed promoter complex. The results suggest that one functional state of the nucleotide-bound activator serves to alter DNA binding by sigma54 and sigma54-RNA polymerase and appears to drive events that precede DNA opening. Clear evidence for a DNA-interacting activity in the AAA+ domain of PspF was obtained, suggesting that PspF may make a direct contact to the DNA component of a basal promoter complex to promote changes in sigma54-RNA polymerase-DNA interactions that favour open complex formation. We also provide evidence for two distinct closed promoter complexes with differing stabilities. PMID- 15333693 TI - Relative gene-silencing efficiencies of small interfering RNAs targeting sense and antisense transcripts from the same genetic locus. AB - Short interfering RNAs (siRNAs) directed against different regions of genes display marked variation in their potency in mediating mRNA degradation. Various factors have been proposed to affect the efficacy of siRNA. We explored some of the factors by evaluating in cultured human cells 28 randomly selected siRNAs targeting the GPR39 and MGC29643 transcripts derived from the same genetic locus but transcribed in opposite directions. Twenty of the 24 siRNAs targeting the overlapping regions of the transcripts simultaneously reduced the levels of both transcripts. Single nucleotide changes in either of the siRNA strands significantly reduced the gene-silencing efficiency of the siRNA on targeted sense transcript without affecting the antisense transcript. Overall, we observed a greater gene-silencing efficiency on the MGC29643 transcript than on the GPR39 transcript in HeLa cells. Since MGC29643 transcript is more abundant than the GPR39 transcript [0.24 versus 0.008% relative to 100% for glyceraldehyde-3 phosphate dehydrogenase (GAPDH)], the results suggest that the abundance of the mRNA affects the efficiency of silencing. Two additional observations supported this hypothesis. First, GAPDH whose intracellular level is the highest of the three was the most efficiently silenced. Second, a reversal of gene-silencing efficiency was observed in U-138 MG cells in which the relative abundance of the GPR39 and MGC29643 transcripts is also reversed. Our study suggests that low abundant transcripts are less susceptible to siRNA-mediated degradation than medium- and high-abundant transcripts. PMID- 15333694 TI - Phosphorylation at 5' end of guanosine stretches inhibits dimerization of G quadruplexes and formation of a G-quadruplex interferes with the enzymatic activities of DNA enzymes. AB - During an analysis of DNA enzymes by gel electrophoresis, we found that some DNA enzymes can adopt more than one conformation. The DNA enzyme Dz31 that formed more than one conformer contained a stretch of G residues. Further investigations, involving kinetic analysis and measurements of circular dichroism, indicated that this DNA enzyme and its derivatives formed G quadruplexes. Moreover, we found that some derivative oligomers were capable of forming dimeric G-quadruplexes. We also compared the catalytic activities of Dz31 and its mutant derivatives. The present findings suggest that DNA enzymes with five or more continuous G residues are less favorable than those without G5 in the association step in the enzymatic reaction and, thus, the choice of targets that contain a continuous stretch of C residues downstream of the cleavage site should be avoided. In addition, we found that negative charge-charge repulsion disrupted the dimerization of G-quadruplexes when a phosphate group was added directly to the 5'-terminal G of oligomers with continuous guanosine residues. In the case of 5'-phosphorylated G5CTA, direct attachment of a phosphate group to the continuous G5 sequence inhibited dimerization of G-quadruplexes, at least during electrophoresis on a denaturing gel. PMID- 15333695 TI - Rational design of DNA sequences for nanotechnology, microarrays and molecular computers using Eulerian graphs. AB - Nucleic acids are molecules of choice for both established and emerging nanoscale technologies. These technologies benefit from large functional densities of 'DNA processing elements' that can be readily manufactured. To achieve the desired functionality, polynucleotide sequences are currently designed by a process that involves tedious and laborious filtering of potential candidates against a series of requirements and parameters. Here, we present a complete novel methodology for the rapid rational design of large sets of DNA sequences. This method allows for the direct implementation of very complex and detailed requirements for the generated sequences, thus avoiding 'brute force' filtering. At the same time, these sequences have narrow distributions of melting temperatures. The molecular part of the design process can be done without computer assistance, using an efficient 'human engineering' approach by drawing a single blueprint graph that represents all generated sequences. Moreover, the method eliminates the necessity for extensive thermodynamic calculations. Melting temperature can be calculated only once (or not at all). In addition, the isostability of the sequences is independent of the selection of a particular set of thermodynamic parameters. Applications are presented for DNA sequence designs for microarrays, universal microarray zip sequences and electron transfer experiments. PMID- 15333696 TI - Novel non-coding RNAs in Dictyostelium discoideum and their expression during development. AB - The quest for non-coding RNAs (ncRNAs) in the last few years has revealed a surprisingly large number of small RNAs belonging to previously known as well as entirely novel classes. Computational and experimental approaches have uncovered new ncRNAs in all kingdoms of life. In this work, we used a shotgun cloning approach to construct full-length cDNA libraries of small RNAs from the eukaryotic model organism Dictyostelium discoideum. Interestingly, two entirely novel classes of RNAs were identified of which one is developmentally regulated. The RNAs within each class share conserved 5'- and 3'-termini that can potentially form stem structures. RNAs of both classes show predominantly cytoplasmic localization. In addition, based on conserved structure and/or sequence motifs, several of the identified ncRNAs could be divided into classes known from other organisms, e.g. 18 small nucleolar RNA candidates (17 box C/D, of which a few are developmentally regulated, and one box H/ACA). Two ncRNAs showed a high degree of similarity to the small nuclear U2 RNA and signal recognition particle RNA (SRP RNA), respectively. Furthermore, the majority of the regions upstream of the sequences encoding the isolated RNAs share conserved motifs that may constitute new promoter elements. PMID- 15333697 TI - Design and synthesis of fluorescent substrates for human tyrosyl-DNA phosphodiesterase I. AB - Tyrosyl-DNA phosphodiesterase 1 (Tdp1) is a DNA repair enzyme that acts upon protein-DNA covalent complexes. Tdp1 hydrolyzes 3'-phosphotyrosyl bonds to generate 3'-phosphate DNA and free tyrosine in vitro. Mutations in Tdp1 have been linked to patients with spinocerebellar ataxia, and over-expression of Tdp1 results in resistance to known anti-cancer compounds. Tdp1 has been shown to be involved in double-strand break repair in yeast, and Tdp1 has also been implicated in single-strand break repair in mammalian cells. Despite the biological importance of this enzyme and the possibility that Tdp1 may be a molecular target for new anti-cancer drugs, there are very few assays available for screening inhibitor libraries or for characterizing Tdp1 function, especially under pre-steady-state conditions. Here, we report the design and synthesis of a fluorescence-based assay using oligonucleotide and nucleotide substrates containing 3'-(4-methylumbelliferone)-phosphate. These substrates are efficiently cleaved by Tdp1, generating the fluorescent 4-methylumbelliferone reporter molecule. The kinetic characteristics determined for Tdp1 using this assay are in agreement with the previously published values, and this fluorescence-based assay is validated using the standard gel-based methods. This sensitive assay is ideal for kinetic analysis of Tdp1 function and for high-throughput screening of Tdp1 inhibitory molecules. PMID- 15333698 TI - Enzymatic switching for efficient and accurate translesion DNA replication. AB - When cyclobutane pyrimidine dimers stall DNA replication by DNA polymerase (Pol) delta or epsilon, a switch occurs to allow translesion synthesis by DNA polymerase eta, followed by another switch that allows normal replication to resume. In the present study, we investigate these switches using Saccharomyces cerevisiae Pol delta, Pol epsilon and Pol eta and a series of matched and mismatched primer templates that mimic each incorporation needed to completely bypass a cis-syn thymine-thymine (TT) dimer. We report a complementary pattern of substrate use indicating that enzymatic switching involving localized translesion synthesis by Pol eta and mismatch excision and polymerization by a major replicative polymerase can account for the efficient and accurate dimer bypass known to suppress sunlight-induced mutagenesis and skin cancer. PMID- 15333699 TI - Maternal nutrition and fetal development. AB - Nutrition is the major intrauterine environmental factor that alters expression of the fetal genome and may have lifelong consequences. This phenomenon, termed "fetal programming," has led to the recent theory of "fetal origins of adult disease." Namely, alterations in fetal nutrition and endocrine status may result in developmental adaptations that permanently change the structure, physiology, and metabolism of the offspring, thereby predisposing individuals to metabolic, endocrine, and cardiovascular diseases in adult life. Animal studies show that both maternal undernutrition and overnutrition reduce placental-fetal blood flows and stunt fetal growth. Impaired placental syntheses of nitric oxide (a major vasodilator and angiogenesis factor) and polyamines (key regulators of DNA and protein synthesis) may provide a unified explanation for intrauterine growth retardation in response to the 2 extremes of nutritional problems with the same pregnancy outcome. There is growing evidence that maternal nutritional status can alter the epigenetic state (stable alterations of gene expression through DNA methylation and histone modifications) of the fetal genome. This may provide a molecular mechanism for the impact of maternal nutrition on both fetal programming and genomic imprinting. Promoting optimal nutrition will not only ensure optimal fetal development, but will also reduce the risk of chronic diseases in adults. PMID- 15333700 TI - ATB0/ASCT2 expression in residual rabbit bowel is decreased after massive enterectomy and is restored by growth hormone treatment. AB - Two weeks after 70% enterectomy, glutamine (Gln) transport is downregulated in rabbit residual bowel due to a decrease in system B(0) activity. Providing epidermal growth factor (EGF) and growth hormone (GH) restores Gln transport by increasing systems A and B(0,+) activities. We hypothesized that changes in Na(+) dependent broad-spectrum neutral amino acid transporter (ATB(0)/ASCT2) protein and mRNA expression correlate with system B(0) activity. New Zealand White rabbits underwent 70% jejunoileal resection or no resection. Resected rabbits immediately received parenteral EGF, GH, both, or neither agent for 2 wk. Tissues harvested from jejunum, ileum, and colon were subjected to Western and Northern blot analyses for ATB(0)/ASCT2 protein and mRNA. In all tissues, ATB(0)/ASCT2 mRNA was reduced by approximately 50% in resected rabbits compared with nonresected controls. Similar reductions in protein amount occurred in the ileum and cecum. None of the growth factor treatments restored ATB(0)/ASCT2 protein, but GH treatment increased ATB(0)/ASCT2 mRNA abundance 250% in the residual ileum. Because changes in the ATB(0)/ASCT2 protein amount paralleled those in the system B(0) activity in this model, it is likely that this is the protein responsible for this transport system. The increase in mRNA abundance in rabbits treated with GH for 2 wk may be a harbinger of subsequent increases in transporter protein and activity. Unlike reported upregulation of transporters in human colon after small bowel resection, ATB(0)/ASCT2 protein and mRNA expression in rabbit colon are decreased, suggesting different regulatory pathways. PMID- 15333701 TI - Kinetic parameters and plasma zinc concentration correlate well with net loss and gain of zinc from men. AB - The search for a reliable, convenient indicator of Zn status was the focus of research for several decades. Plasma Zn concentration is still the most widely used clinical measurement, despite the known problems of interpretation. More recently, researchers suggested that isotopically determined kinetic parameters, such as the exchangeable Zn pool (EZP), may more accurately and reliably reflect body Zn status. The objective of this study was to examine the relationship between net body Zn loss and gain during acute changes in dietary Zn intake with biochemical and kinetic indices of Zn status. Five men participated in an 85-d Zn depletion/repletion study. Net body Zn loss and gain were determined from the difference between dietary plus intravenously administered Zn and Zn excretion. Biochemical indicators of Zn status included plasma Zn, plasma alkaline phosphatase activity, and plasma retinol binding protein concentration. Following intravenous administration of (70)Zn or (67)Zn, a compartmental model was used to determine EZP mass, fractional Zn absorption, endogenous zinc excretion (EZE), and plasma Zn flux. The changes in total body zinc correlated best with changes in plasma Zn (r(2) = 0.826, P < 0.001), EZE (r(2) = 0.773, P < 0.001), and plasma Zn flux (r(2) = 0.766, P < 0.001). This study confirms that plasma Zn concentration is a valid indicator of whole-body Zn status in the absence of confounding factors; however, further research is needed to determine how kinetic parameters respond to conditions where plasma Zn concentration is known to be unreliable. PMID- 15333702 TI - The amino acid need for milk synthesis is defined by the maximal uptake of plasma amino acids by porcine mammary glands. AB - To define dietary indispensable amino acid (IAA) needs for milk synthesis by the mammary glands (MG), 16 lactating sows were fed 1 of 4 isocaloric diets varying in protein concentrations (from 78 to 235 g/kg) with an ideal amino acid (AA) pattern. On d 9, 13, 17, and 21 of lactation, blood samples were obtained simultaneously from a carotid artery and the main mammary vein every 30 min over 6 h. A quadratic regression model of the log mammary arteriovenous difference (AVD) of plasma IAA (y) against daily intake of dietary IAA (X) was established. First, the reverse log intercept, defined as the mammary AVD at zero dietary AA supply, was used to quantify the contribution of endogenous IAA. The quantification was validated by body N balance coupled with AA composition analysis. Then, the estimated vertex (y(max), X(i)) was used in 2 aspects: 1) The maximal mammary uptake of plasma IAA, quantified by multiplying the maximal mammary AVD and plasma flow rate, was considered the physiological IAA need for milk synthesis. 2) Corresponding to the y(max), dietary IAA intake (X(i)) would represent the total dietary IAA requirement, i.e., the sum of maintenance need and milk synthesis need after adjustment for body weight loss. Thus, dietary IAA needs for milk synthesis were derived. Moreover, the estimate of lysine need for milk synthesis in this study was identical to an estimate obtained from multiple regression analysis of feeding trial data. We conclude that dietary IAA needs for milk synthesis can be quantified by the maximal uptakes of plasma IAA by porcine MG. PMID- 15333703 TI - Restriction of dietary energy and protein induces molecular changes in young porcine skeletal muscles. AB - Little is known about the molecular changes in response to dietary restriction (energy and/or protein) in young growing skeletal muscles. To profile such changes and to gain insights into the signaling molecules that could mediate the diet effects, a dedicated porcine skeletal muscle cDNA-microarray approach was used to characterize differential muscle gene expression between conventionally fed and diet-restricted (20% less protein and 7% less energy) growing pigs, reared from 9 to 21 wk of age. In both red and white muscles, diet restriction resulted in the accumulation of significantly more intramuscular fat, and in the increased expression of genes involved in substrate (protein, glycogen, and lipid) turnover, in translation and mitochondrial function, and in raising glycolytic and oxidative phosphorylation potentials. The unexpected increase in intramuscular lipids in diet-restricted growing pigs could have important health implications for restricted diets in childhood. Despite reduced circulating insulin, more genes, including several novel growth modulatory genes, had higher expression levels, indicating that the cellular response to dietary restriction is an active process. One such responsive gene, P311, was most highly expressed in striated muscles and had a differentiation-dependent increase of expression in murine C2C12 cells, suggesting a role in differentiation/postdifferentiation phenotype determination. PMID- 15333704 TI - CYP7A1 A-278C polymorphism affects the response of plasma lipids after dietary cholesterol or cafestol interventions in humans. AB - The response of plasma lipids to dietary cholesterol and fat varies among individuals. Variations in genes involved in cholesterol metabolism can be important in these interindividual differences. The rate-limiting enzyme in the conversion of cholesterol into bile acids is cholesterol 7alpha-hydroxylase (CYP7A1). We investigated the effect of the A278-C promoter polymorphism in the CYP7A1 gene on responses of plasma lipids to an increased intake in dietary cholesterol (742 +/- 114 mg/d), cafestol (57 +/- 6 mg/d), saturated fat [change of 8-9 energy percent/d (en%/d)] and trans fat (change of 10-11 en%/d) in 496 normolipidemic subjects. These responses were measured in 26 previously published dietary trials. After adjustment for the apolipoprotein E genotype effect, AA subjects consuming a cholesterol-rich diet had a smaller increase in plasma HDL cholesterol than CC-subjects (0.00 +/- 0.02 vs. 0.17 +/- 0.04 mmol/L; P < 0.001). Upon intake of cafestol, AA-subjects had a smaller increase in plasma total cholesterol than CC-subjects (0.69 +/- 0.10 vs. 1.01 +/- 0.10 mmol/L; P = 0.028). No effects of the polymorphism were found in the saturated and trans fat interventions. In conclusion, the CYP7A1 polymorphism has a small but significant effect on the increase in plasma HDL cholesterol and plasma total cholesterol after an increased intake of dietary cholesterol and cafestol, respectively. PMID- 15333705 TI - Starvation and feeding a high-carbohydrate, low-fat diet regulate the expression sterol regulatory element-binding protein-1 in chickens. AB - In mammalian liver, the mature form of sterol regulatory element-binding protein 1c (SREBP-1c) is an important activator of a wide array of genes involved in triacylglycerol biosynthesis. Starvation and feeding a high-carbohydrate, low-fat diet modulate the concentration of mature SREBP-1c primarily by a pretranslational mechanism. It is not known whether alterations in nutritional status regulate the concentration of SREBPs in nonmammalian species. In this study, we found that in previously starved chicks, feeding a high-carbohydrate, low-fat diet stimulated a robust increase (14-fold at 5 h of feeding) in the concentration of mature SREBP-1 in liver. Feeding a high-carbohydrate, low-fat diet also increased the concentration of precursor SREBP-1 and SREBP-1 messenger RNA in chick liver; however, the magnitude of this effect was substantially lower than that observed for mature SREBP-1. DNA binding experiments demonstrated that 3 protein complexes containing SREBP bound the acetyl-CoA carboxylase-alpha (ACCalpha) sterol regulatory element (SRE) in chick liver and that the binding activity of 2 of these complexes was increased by consumption of a high carbohydrate, low-fat diet. Additional analyses showed that feeding a high carbohydrate, low-fat diet had no effect on the concentration of mature SREBP-2 and the binding of SREBP-2 to the ACCalpha SRE in chick liver. These results indicate that alterations in the concentration of mature SREBP-1 play a role in mediating the effects of starvation and feeding a high-carbohydrate, low-fat diet on ACCalpha transcription in chick liver and that diet-induced changes in mature SREBP-1 concentration in chick liver are mediated primarily by a posttranslational mechanism. PMID- 15333706 TI - The diurnal rhythm of the intestinal transporters SGLT1 and PEPT1 is regulated by the feeding conditions in rats. AB - The intestinal Na(+)/glucose cotransporter 1 (SGLT1) and H(+)/peptide cotransporter 1 (PEPT1) play important roles in the absorption of carbohydrate and protein. Although they exhibit a diurnal rhythm in their expression and function, the factors responsible for this are unclear. In the present study, we examined the effects of various feeding conditions on the diurnal rhythm of intestinal SGLT1 and PEPT1. Rats were divided into 1 of 4 groups: group 1 was fed, group 2 was food deprived for 1-4 d, group 3 was food deprived for 4 d and then refed for 1 or 2 d, and group 4 was fed during the daytime (0900-1500 h) for 10 d. In fed rats, the SGLT1 protein level was significantly higher at 2000 h than at 0800 h. However, in rats deprived of food for 2-4 d, protein levels did not differ between 0800 and 2000 h. In contrast, the SGLT1 messenger RNA (mRNA) level was significantly higher at 2000 h than at 0800 h in rats deprived of food for 4 d. Refeeding for 2 d after 4 d of food deprivation returned the diurnal variation in SGLT1 and PEPT1 protein expressions to normal. Consuming food during the daytime only shifted the peaks of SGLT1 and PEPT1 mRNAs and protein expressions from the dark phase to the light phase. These findings suggest that food intake, rather than the light cycle, greatly affects the diurnal rhythm of SGLT1 and PEPT1 expressions. PMID- 15333707 TI - Gender and sodium-ascorbate transporter isoforms determine ascorbate concentrations in mice. AB - We evaluated the hypothesis that sodium-dependent vitamin C (ascorbate) transporters SVCT1 and SVCT2 (encoded by genes Slc23a1 and Slc23a2) regulate ascorbate concentrations in tissues of adult mice. Slc23a2+/- and Slc23a2+/+ mice were fed an ascorbate-free diet for 10-12 wk, and then segregated according to gender and genome, and were placed in groups of 3-4 in metabolic cages for 24-h urine collection. RT-PCR analysis showed that liver and kidney expressed mainly SVCT1, and brain, skeletal muscle, and spleen expressed predominantly SVCT2. Slc23a2+/- mice had low SVCT2 but normal SVCT1 messenger RNA (mRNA) levels, which did not differ between genders. Ascorbate concentrations were lower in Slc23a2+/- than Slc23a2+/+ mice in tissues where SVCT2 was the main isoform. Compared with males, females had lower ascorbate excretion and ascorbate:creatinine ratio in urine and had higher ascorbate concentrations in plasma and SVCT1-predominant tissues. SVCT2 contributed to a gender effect in spleen because males had higher spleen ascorbate concentration than females in wild-type but not in Slc23a2+/- mice. Hepatic gulonolactone oxidase mRNA and activity levels did not differ with genotype or gender, indicating no differences in ascorbate synthesis. We concluded that SVCT2 is a major determinant of ascorbate accumulation in tissues lacking SVCT1. The SVCT isoforms appear to function independently of one another because SVCT1 expression and ascorbate concentrations in SVCT1-predominant organs were not affected by SVCT2 deficiency. Additionally, lower ascorbate excretion in females may elevate the vitamin's concentrations in plasma and tissues expressing SVCT1 that, unlike SVCT2, is not saturated by plasma ascorbate concentrations. PMID- 15333708 TI - Biotin supplementation increases expression of the cytochrome P450 1B1 gene in Jurkat cells, increasing the occurrence of single-stranded DNA breaks. AB - DNA microarray studies provided evidence that biotin supplementation increases the abundance of mRNA encoding cytochrome P(450) 1B1 (CYP1B1) in human lymphocytes. CYP1B1 hydroxylates procarcinogens, generating electrophilic mutagens. Here, we sought to identify the signaling pathways that increase the expression of CYP1B1 in biotin-supplemented human T (Jurkat) cells and to determine whether activation of the CYP1B1 gene is associated with increased occurrence of single-stranded DNA breaks. Jurkat cells were cultured in biotin deficient (0.025 nmol/L) and biotin-supplemented (10 nmol/L) media. The transcriptional activity of a CYP1B1 reporter gene construct was 24% greater in biotin-supplemented compared with biotin-deficient cells (P < 0.01). Similarly, the abundance of CYP1B1 mRNA was 72% greater in biotin-supplemented than in biotin-deficient cells (P < 0.05). Electrophoretic mobility shift assays suggested that Sp1 sites in the regulatory region of the CYP1B1 gene play important roles in transcriptional activation by biotin. The abundance of CYP1B1 protein and activity of CYP1B1 were 124 and 35% greater, respectively, in biotin supplemented compared with biotin-deficient cells (P < 0.05). The increased expression of CYP1B1 in biotin-supplemented cells was associated with an increase in the occurrence of single-stranded DNA breaks compared with biotin-deficient cells; synthetic inhibitors of CYP1B1 prevented strand breaks, suggesting that the effects of biotin were specific for CYP1B1. These studies provide evidence that transcription factors with an affinity for Sp1 sites mediate transcriptional activation of the CYP1B1 gene in biotin-supplemented T cells, increasing the occurrence of single-stranded DNA breaks. PMID- 15333709 TI - Sulforaphane inhibits human MCF-7 mammary cancer cell mitotic progression and tubulin polymerization. AB - Sulforaphane (SUL), an isothiocyanate derived from hydrolysis of glucoraphanin in broccoli and other cruciferous vegetables, was shown to induce phase II detoxification enzymes, inhibit chemically induced mammary tumors in rodents, and more recently, to induce cell cycle arrest and apoptosis in colon cancer cells. In the present study, we demonstrate that SUL also acts to inhibit proliferation of MCF-7 adenocarcinoma cells from the human breast. Treatment of synchronized MCF-7 cells with 15 micromol/L SUL resulted in significant (P < 0.05) G(2)/M cell cycle arrest (167% of control) and elevated cyclin B1 protein (175% of control) within 24 h. Moreover, 15 micromol/L SUL significantly (P < 0.05) induced phosphorylation of histone H1 (167% of control), blocked cells in early mitosis ( approximately 10-fold increase over control), and disrupted polymerization of mitotic microtubules in vivo. Subsequent exposure of purified bovine brain tubulin to relatively high doses of SUL significantly (P < 0.05) inhibited both tubulin polymerization rate (51% of control) and total tubulin polymerization (78% of control) in vitro. Additionally, polymerization of purified tubulin exposed to isothiocyanate-containing analogs of SUL was similarly inhibited. Taken together, these findings indicate that SUL has mammary cancer suppressive actions involving mitotic cell cycle arrest and suggest a mechanism linked to the disruption of normal tubulin polymerization and/or more subtle effects on microtubule dynamics. PMID- 15333710 TI - An epoxide-furanoid rearrangement of spinach neoxanthin occurs in the gastrointestinal tract of mice and in vitro: formation and cytostatic activity of neochrome stereoisomers. AB - Neoxanthin, a major carotenoid in green leafy vegetables, was reported to exhibit potent antiproliferative effect via apoptosis induction on human prostate cancer cells. However, the metabolic fate of dietary neoxanthin in mammals remains unknown. In the present study, we investigated the gastrointestinal metabolism of neoxanthin in mice and the in vitro digestion of spinach, and estimated the antiproliferative effect of neoxanthin metabolites on PC-3 human prostate cancer cells. Two hours after the oral administration to mice of purified neoxanthin, unchanged neoxanthin and stereoisomers of neochrome (8'-R/S) were detected in the plasma, liver, and small intestinal contents. To estimate the effect of intragastric acidity on the conversion of dietary neoxanthin into neochrome (epoxide-furanoid rearrangement), spinach was digested in vitro by incubating it with a pepsin-HCl solution at pH 2.0 or 3.0 (gastric phase) followed by a pancreatin-bile salt solution (intestinal phase). Spinach neoxanthin was largely converted into (R/S)-neochrome during the digestion when the gastric phase was set at pH 2.0, whereas the rearrangement was observed to a lesser extent at pH 3.0. (R/S)-neochrome dose-dependently inhibited the proliferation of PC-3 cells as well as neoxanthin at concentrations < or = 20 micromol/L. Although neoxanthin induced evident apoptotic cell death, (R/S)-neochrome inhibited the cell proliferation without obvious apoptosis induction. These results indicate that dietary neoxanthin is partially converted into (R/S)-neochrome by intragastric acidity before intestinal absorption and that (R/S)-neochrome exhibits an antiproliferative effect on PC-3 cells by the induction of cytostasis. PMID- 15333711 TI - 1alpha,25-dihydroxycholecalciferol increases the expression of vascular endothelial growth factor in C3H10T1/2 mouse embryo fibroblasts. AB - Evidence suggests that biologically active vitamin D, 1,25 dihydroxycholecalciferol [1,25(OH)(2)D(3)], may inhibit carcinogenesis. Because angiogenesis is crucial to carcinogenesis, 1,25(OH)(2)D(3) regulation of proangiogenic vascular endothelial growth factor (VEGF) secretion was investigated in cellular models for multistage carcinogenesis. Conditioned media from 1,25(OH)(2)D(3)-treated C3H10T(1/2) mouse fibroblasts and their Harvey ras oncogene transfected counterparts (rasneo11a cells) induced human umbilical vein endothelial cell (HUVEC) proliferation (1.3 and 0.3 times, respectively, P < 0.05), suggesting that 1,25(OH)(2)D(3) altered the angiogenic phenotype of the cells. Although rasneo11a cells secreted less VEGF than C3H10T(1/2) cells (97%, P < 0.005), 1,25(OH)(2)D(3) induced C3H10T(1/2) and rasneo11a cells to secrete 2 and 3 times, respectively, more VEGF than controls (P < 0.05). Similar effects on VEGF release occurred after 1,25(OH)(2)D(3) treatment of MCF10A and MCF10Aras cells, a human breast epithelial cell model for multistage carcinogenesis. In C3H10T(1/2) cells, 1,25(OH)(2)D(3) activated the VEGF promoter in a dose dependent (5-100 nmol/L) manner (maximum 60%) and all doses induced VEGF secretion (P < 0.05). 1,25(OH)(2)D(3) induced VEGF mRNA expression ( approximately 50%) from 2 through 24 h; VEGF release was significantly increased at 8 h and sustained for 24 h. VEGF mRNA expression and release declined as C3H10T(1/2) cells grew more confluent, whereas the magnitude of 1,25(OH)(2)D(3) stimulated changes in VEGF was greater in confluent (3.3 times RNA; 3.5 times release) than in subconfluent (50% RNA; 100% release) cultures (P < 0.05). Thus, 1,25(OH)(2)D(3) increases VEGF secretion, and in C3H10T(1/2) cells, this is likely through activation of the VEGF promoter and induction of gene expression. These data contribute to understanding the role 1,25(OH)(2)D(3) plays in regulation of angiogenesis in normal compared with disease states. PMID- 15333712 TI - Dietary iron affects inflammatory status in a rat model of colitis. AB - Iron deficiency anemia is a common feature in inflammatory bowel disease, and oral supplementation is one of the mainstay therapies. However, there is some concern that oral iron supplementation may lead to oxidative stress and exacerbation of inflammation. Our objective was to study the effect of severely deficient, moderately deficient, normal and high iron status on oxidative stress and the course of inflammation in a rat model of colitis induced by 2,4,6 trinitrobenzene sulfonic acid (TNBS). The rats were randomly assigned to receive the low-iron diet for 3 (moderately iron-deficient group, n = 16) or 5 (severely iron-deficient group, n = 16) wk, the normal iron diet for 2 wk (normal iron group, n = 16) or the high-iron diet for 2 wk (high-iron group, n = 16). Malondialdehyde concentration, electroparamagnetic resonance measurement, myeloperoxidase activity, and histological analysis were used to evaluate oxidative stress. Noncolitic rats in the high-iron group had higher oxidative stress parameters than those in the other groups. The induction of colitis resulted in severe inflammatory changes in the high-iron and severely iron deficient groups, and produced higher histological scores in the colon of the normal and high-iron groups. Iron overload, oxidative stress, and inflammation were lower in the moderately iron-deficient group compared with the other 3 groups. In conclusion, we suggest that low rather than normal or high iron supplementation should be considered for the treatment of iron deficiency in inflammatory bowel disease. PMID- 15333713 TI - Weaning induces both transient and long-lasting modifications of absorptive, secretory, and barrier properties of piglet intestine. AB - This study investigated intestinal physiology of piglets at weaning. Piglets (n = 60) weaned at 21 d were food deprived for 2 d and then tube-fed using 2 different diets (a conventional diet vs. a wheat-enriched diet). They were slaughtered at d 0, 2, 5, 8, or 15 postweaning. Jejunum, ileum, and colon were mounted in Ussing chambers. In addition, segments of the proximal jejunum of 4 growing pigs were studied 35 d after weaning. Secretory function was assessed by basal short circuit current (Isc) and secretagogue-stimulated Isc. Glucose absorption was measured by the increase in Isc after the addition of glucose. Epithelial barrier function was measured by transmucosal resistance (R) and horseradish peroxidase (HRP) fluxes across the epithelium. There were no significant differences between the pigs fed the 2 diets for any of the parameters studied. As already reported, a transient villous atrophy was observed. At the same time, we observed an increased basal Isc in jejunum and colon, increased glucose absorption and a dramatic drop of R in jejunum. These parameters had returned to preweaning values by d 5. Weaning was also followed by long-lasting modifications. In jejunum, responses to the secretagogues and glucose absorption were decreased at wk 2 after weaning and were not different between d 15 and 35. Ileal transmucosal resistance increased on d 5 and was stable thereafter. HRP flux in jejunum declined on d 2 and stayed at this low level throughout the experiment. We conclude that weaning induces transient dramatic changes in intestinal physiology but is also a period of maturation of the intestine. PMID- 15333714 TI - Cis-9, trans-11 and trans-10, cis-12 conjugated linoleic acid isomers do not modify body composition in adult sedentary or exercised rats. AB - Dietary CLA isomers were shown to reduce adipose tissues in growing animals, mainly in mice, but their effects in adult animals remain unclear. This study was conducted to determine whether these effects depend on the isomer fed, on physical activity, or on the initial level of body fat. Male Wistar rats (4 mo old) were fed for 6 wk diets containing either no CLA, the cis-9, trans-11 CLA isomer (10 g/kg), the trans-10, cis-12 CLA isomer (10 g/kg), or both isomers (10 g/kg each). Half of the rats were assigned to exercise by treadmill running (1 h/d, 22 m/min). The initial body fat level was normal (12.7%) in a first trial, and high (18.9%) in a second trial. Chemical and anatomical body compositions were determined by chemical analysis and organ dissection. In both trials, the CLA diets, whatever the isomer, had no effect on food intake and body weight changes, on body chemical composition (fat, protein and water contents or gains), or on the body anatomical composition (weights or gains in epididymal and perirenal adipose tissues, in liver and in 4 muscles). There was no interaction between CLA treatment and physical activity. In conclusion, adult male rats do not appear to be responsive to the fat-to-lean partitioning effect of CLA described in growing rats. This was not affected by exercise or initial body fat level. PMID- 15333715 TI - Iron absorption from NaFeEDTA is downregulated in iron-loaded rats. AB - NaFeEDTA is a promising fortificant for use in plant foods, because it is less susceptible to iron absorption inhibitors and has fewer undesirable impacts on sensory quality than ferrous sulfate. However, the hypothesis that iron absorption from NaFeEDTA is effectively downregulated in iron-overload conditions has not been thoroughly tested. Therefore, the objective of this study was to compare downregulation of iron absorption from ferrous sulfate and NaFeEDTA in intact iron-loaded rats. Male Sprague-Dawley rats were fed diets containing either ferrous sulfate (35 mg Fe per 1 kg diet) or elemental iron (30,000 mg Fe per 1 kg diet) for 29 d to achieve basal or iron-loaded status. While body weights and hemoglobin concentrations were the same for basal and iron-loaded rats, nonheme-iron concentrations in liver, spleen, and kidney were all significantly higher in iron-loaded rats, indicating elevated iron status. Percentage of iron absorption from (59)Fe-labeled ferrous sulfate and NaFeEDTA, determined from whole-body retention of (59)Fe activity, was 64.7 and 49.4% in basal rats but decreased to 12.8 and 10.2% in iron-loaded rats, respectively. The reductions in percentage of iron absorption from both iron sources in rats as a result of iron loading were comparable (about -80% for both iron sources). Our results suggest that iron absorption from NaFeEDTA and ferrous sulfate is downregulated to a similar extent in iron-loaded rats. Hence, NaFeEDTA is no more likely than ferrous sulfate to exacerbate iron overload in subjects with adequate body iron stores. PMID- 15333716 TI - Anthocyanins are efficiently absorbed from the small intestine in rats. AB - Anthocyanins are natural pigments that possess antioxidant activities and are implicated in various health effects. Recent studies showed that the stomach is a site of anthocyanin absorption. However, the fate of anthocyanins in the small intestine remains unknown. We therefore investigated anthocyanin absorption after in situ perfusion of the jejunum + ileum in rats. The intestine was perfused for 45 min with a physiological buffer supplemented with various anthocyanins. Purified anthocyanin glycosides (9.2 nmol/min) or blackberry (9.0 nmol/min) or bilberry (45.2 nmol/min) anthocyanins were perfused. A high proportion of anthocyanin glycosides was absorbed through the small intestine after perfusion. The rate of absorption was influenced by the chemical structure of the anthocyanin and varied from 10.7 (malvidin 3-glucoside) to 22.4% (cyanidin 3 glucoside). Regardless of the anthocyanins perfused, only glycosides were recovered in the intestinal lumen. After perfusion of a high amount of blackberry anthocyanins (600 nmol/min), native cyanidin 3-glucoside was recovered in urine and plasma from the aorta and mesenteric vein. Methylated and/or glucuronidated derivatives were also identified. Analysis of bile samples revealed that cyanidin 3-glucoside and its methylated derivatives (peonidin 3-glucoside + peonidin glucuronide) quickly appeared in bile. This study demonstrated that anthocyanin glycosides are rapidly and efficiently absorbed from the small intestine. Furthermore, anthocyanins are quickly metabolized and excreted into bile and urine as intact glycosides as well as methylated forms and glucuronidated derivatives. PMID- 15333717 TI - Assessment of lutein bioavailability from meals and a supplement using simulated digestion and caco-2 human intestinal cells. AB - Lutein and zeaxanthin are selectively accumulated in the lens and macular region of the retina. It was suggested that these xanthophylls protect ocular tissues from free-radical damage that can cause cataracts and age-related macular degeneration. Insights regarding the absorption of dietary xanthophylls for delivery to ocular tissues are limited. Our primary objective was to examine factors affecting the transfer of lutein from foods to absorptive intestinal epithelial cells during digestion. Lutein and other carotenoids present in spinach puree and lutein from a commercial supplement were relatively stable during in vitro digestion. Micellarization of lutein and zeaxanthin during the small intestinal phase of digestion exceeded that of beta-carotene and was greater for xanthophylls in oil-based supplements than in spinach. Apical uptake of lutein from micelles by Caco-2 human intestinal cells was linear for at least 8 h, and accumulation from synthetic micelles exceeded that from micelles generated during simulated digestion. Stimulation of chylomicron synthesis resulted in the secretion of 7.6 +/- 0.1% of cellular lutein into the triglyceride-rich fraction in the basolateral chamber. These data support the use of simulated digestion and the Caco-2 cell model as effective tools for identifying factors affecting absorption of dietary carotenoids. PMID- 15333718 TI - Growth potential, but not body weight or moderate limitation of lysine intake, affects inevitable lysine catabolism in growing pigs. AB - Inevitable catabolism contributes to the inefficiency of using dietary lysine intake for body protein deposition (PD). This study was conducted to determine the effects of true ileal digestible (TID) lysine intake, body weight (BW), and growth potential on lysine catabolism in growing pigs. Starting at 15 kg BW, 16 female Yorkshire pigs were offered a purified diet providing all nutrients in excess of requirements for maximum protein deposition (PDmax). At approximately 25 kg BW, the pigs' PDmax was determined using the N-balance method. Thereafter, 4 pigs were allocated to each of 4 diets, first-limiting in lysine, providing lysine intakes corresponding to 60, 70, 80, and 90% of estimated requirements for PDmax. The pigs were surgically fitted with catheters in the jugular and femoral veins. Lysine catabolism was determined at 2 BW (40-45 kg, low; 70-75 kg, high) either directly (oxidation) using a primed, constant infusion of l-[1-(14)C] lysine or indirectly (disappearance) using the N-balance method. There was no effect of BW on the rate (g/d) or fraction of TID lysine intake catabolized. Lysine catabolism decreased with increasing growth potential. Lysine disappearance and lysine oxidation (% of TID lysine intake) were independent of lysine intake, except for the lowest lysine intake level, where they were lower. When lysine catabolism was independent of intake, lysine oxidation based on plasma free lysine specific radioactivity (SRA) was lower (9.9% of TID intake) than lysine disappearance (17.4% of TID intake) or lysine oxidation based on liver free lysine SRA (13.4% of TID intake). PMID- 15333719 TI - In vivo imaging of intragastric gelation and its effect on satiety in humans. AB - Previous studies indicated that physical characteristics of food influence satiety, but the relative importance of the oral, gastric, and intestinal behaviors of the food is unclear. The aim of this study was to investigate the satiating effects of 2 types of alginates, which gel weakly or strongly on exposure to acid, compared with guar gum whose viscosity is unaffected by acid. Subjects (n = 12; 3 men, 9 women) ingested a 325-mL sweetened, milk-based meal replacer beverage on 4 separate occasions, either alone as a control or including 1% by weight alginate or guar gum. Intragastric gelling, gastric emptying, and meal dilution were assessed by serial MRI while satiety was recorded for 4 h. MR images showed that all of the meals became heterogeneous in the stomach except for guar, which remained homogeneous. The alginate meals formed lumps in the stomach, with the strong-gelling alginate producing the largest volume. Although gastric emptying was similar for all 4 meals, the sense of fullness at the same gastric volume was significantly greater for all 3 viscous meals than for the control. Compared with the control meal, the strong-gelling alginate (P = 0.031) and guar (P = 0.041) meals increased fullness at 115 min, and the strong-gelling alginate decreased hunger by the 115-min (P = 0.041) and 240-min (P = 0.041) time points. Agents that gel on contact with acid may be useful additions to weight reducing diets. We hypothesize that this effect is due to distension in the gastric antrum and/or altered transport of nutrients to the small intestine in the lumps. PMID- 15333720 TI - Hyperphagia contributes to the normal body composition and protein-energy balance in HIV-infected asymptomatic men. AB - Wasting can occur at an early stage of HIV infection. Both reduced energy intake and increased resting energy expenditure (REE) have been considered as factors in wasting with predominant lean body mass loss, suggesting disturbances of protein metabolism. Our aim was to study protein-energy metabolism in relation to body composition and oral energy intake in asymptomatic patients with HIV infection but receiving no active antiretroviral therapy. Stable-weight asymptomatic male patients (n = 8) at stage A of HIV infection with a detectable viral load were compared with 9 healthy control men. Protein metabolism was studied in the postabsorptive state using a primed constant infusion of l-[1-(13)C]leucine and l [2-(15)N]glutamine. REE was studied by indirect calorimetry, body composition by bioelectrical impedance, and energy intake by dietary records. BMI and lean body mass did not differ between patients and controls. In HIV-infected subjects, energy intake, protein breakdown, protein synthesis, and REE were 57% (P < 0.05), 18% (P < 0.05), 22% (P < 0.05) and 14% (P < 0.05) greater than in controls, respectively. REE and protein breakdown were correlated (r = 0.73, P < 0.05). The hormonal profile was normal in HIV-infected subjects with the exception of low urinary C-peptide and plasma reverse triiodothyronine. Plasma interleukin-6 and tumor necrosis factor-alpha were greater than in controls, but energy intake was 1.53 times the REE in the HIV-infected men. Thus, at the asymptomatic stage of HIV infection, increased protein turnover contributes to the increase in the REE. Moderate hyperphagia, which occurred despite increased levels of cytokines, in conjunction with increased protein synthesis maintains a normal body composition, without significant loss of lean body mass. PMID- 15333721 TI - Maturation of visual acuity is accelerated in breast-fed term infants fed baby food containing DHA-enriched egg yolk. AB - Between 6 and 12 mo of age, blood levels of the (n-3) long-chain PUFA, docosahexaenoic acid (DHA), in breast-fed infants typically decrease due to diminished maternal DHA stores and the introduction of DHA-poor solid foods displacing human milk as the primary source of nutrition. Thus, we utilized a randomized, clinical trial format to evaluate the effect of supplemental DHA in solid foods on visual development of breast-fed infants with the primary outcome, sweep visual-evoked potential (VEP) acuity, as an index for maturation of the retina and visual cortex. At 6 mo of age, breast-fed infants were randomly assigned to receive 1 jar (113 g)/d of baby food containing egg yolk enriched with DHA (115 mg DHA/100 g food; n = 25) or control baby food (0 mg DHA; n = 26). Gravimetric measures were used to estimate the supplemental DHA intake which was 83 mg DHA/d in the supplemented group and 0 mg/d in controls. Although many infants in both groups continued to breast-feed for a mean of 9 mo, RBC DHA levels decreased significantly between 6 and 12 mo (from 3.8 to 3.0 g/100 g total fatty acids) in control infants, whereas RBC DHA levels increased by 34% from 4.1 to 5.5 g/100 g by 12 mo in supplemented infants. VEP acuity at 6 mo was 0.49 logMAR (minimum angle of resolution) and improved to 0.29 logMAR by 12 mo in controls. In DHA-supplemented infants, VEP acuity was 0.48 logMAR at 6 mo and matured to 0.14 logMAR at 12 mo (1.5 lines on the eye chart better than controls). At 12 mo, the difference corresponded to 1.5 lines on the eye chart. RBC DHA levels and VEP acuity at 12 mo were correlated (r = -0.50; P = 0.0002), supporting the need of an adequate dietary supply of DHA throughout 1 y of life for neural development. PMID- 15333722 TI - Olive oils high in phenolic compounds modulate oxidative/antioxidative status in men. AB - The aim of the present study was to evaluate whether olive oils high in phenolic compounds influence the oxidative/antioxidative status in humans. Healthy men (n = 12) participated in a double-blind, randomized, crossover study in which 3 olive oils with low (LPC), moderate (MPC), and high (HPC) phenolic content were given as raw doses (25 mL/d) for 4 consecutive days preceded by 10-d washout periods. Volunteers followed a strict very low-antioxidant diet the 3 d before and during the intervention periods. Short-term consumption of olive oils decreased plasma oxidized LDL (oxLDL), 8-oxo-dG in mitochondrial DNA and urine, malondialdehyde in urine (P < 0.05 for linear trend), and increased HDL cholesterol and glutathione peroxidase activity (P < 0.05 for linear trend), in a dose-dependent manner with the phenolic content of the olive oil administered. At d 4, oxLDL after MPC and HPC, and 8-oxo-dG after HPC administration (25 mL, respectively), were reduced when the men were in the postprandial state (P < 0.05). Phenolic compounds in plasma increased dose dependently during this stage with the phenolic content of the olive oils at 1, 2, 4, and 6 h, respectively (P < 0.01). Their concentrations increased in plasma and urine samples in a dose dependent manner after short-term consumption of the olive oils (P < 0.01). In conclusion, the olive oil phenolic content modulated the oxidative/antioxidative status of healthy men who consumed a very low-antioxidant diet. PMID- 15333723 TI - Blood pressure response to dietary modifications in free-living individuals. AB - A diet rich in fruits, vegetables, and low-fat dairy foods has been shown to lower blood pressure (BP) when all foods are provided. We compared the effect on BP (measured at home) of 2 different self-selected diets: a low-sodium, high potassium diet, rich in fruit and vegetables (LNAHK) and a high-calcium diet rich in low-fat dairy foods (HC) with a moderate-sodium, high-potassium, high-calcium DASH-type diet, high in fruits, vegetables and low-fat dairy foods (OD). Subjects were randomly allocated to 2 test diets for 4 wk, the OD and either LNAHK or HC diet, each preceded by a 2 wk control diet (CD). The changes in BP between the preceding CD period and the test diet period (LNAHK or HC) were compared with the change between the CD and the OD periods. Of the 56 men and 38 women that completed the OD period, 43 completed the LNAHK diet period and 48 the HC diet period. The mean age was 55.6 +/- 9.9 (+/-SD) years. There was a fall in systolic pressure between and the CD and OD [-1.8 +/- 0.5 mm Hg (P < 0.001)]. Compared with OD, systolic and diastolic BPs fell during the LNAHK diet period [-3.5 +/- 1.0 (P < 0.001) and -1.9 +/- 0.7 (P < 0.05) mmHg, respectively] and increased during the HC diet period [+3.1 +/- 0.9 (P < 0.01) and +0.8 +/- 0.6 (P = 0.15) mm Hg, respectively]. A self-selected low-sodium, high-potassium diet resulted in a greater fall in BP than a multifaceted OD, confirming the beneficial effect of dietary intervention on BP in a community setting. PMID- 15333724 TI - Household food insecurity is associated with adult health status. AB - The prevalence of household food security, which reflects adequacy and stability of the food supply, has been measured periodically in the United States and occasionally in high-risk groups or specific regions. Despite a plausible biological mechanism to suggest negative health outcomes of food insecurity, this relation has not been adequately evaluated. This study was conducted in the Lower Mississippi Delta region to examine the association between household food insecurity and self-reported health status in adults. A two-stage stratified cluster sample representative of the population in 36 counties in the Delta region of Arkansas, Louisiana, and Mississippi was selected using list-assisted random digit dialing telephone methodology. After households were selected and screened, a randomly selected adult was interviewed within each sampled household. Data were collected to measure food security status and self-reported mental, physical, and general health status, using the U.S. Food Security Survey Module and the Short Form 12-item Health Survey (SF-12). Data were reported on a sample of 1488 households. Adults in food-insecure households were significantly more likely to rate their health as poor/fair and scored significantly lower on the physical and mental health scales of the SF-12. In regression models controlling for income, gender, and ethnicity, the interaction between food insecurity status and race was a significant predictor of fair/poor health and lower scores on physical and mental health. Household food insecurity is associated with poorer self-reported health status of adults in this rural, high risk sample in the Lower Mississippi Delta. PMID- 15333725 TI - Conditional cash transfers are associated with a small reduction in the rate of weight gain of preschool children in northeast Brazil. AB - Programs providing cash transfers to poor families, conditioned upon uptake of preventive health services, are common in Latin America. Because of the consistent association between undernutrition and poverty, and the role of health services in providing growth promotion, these programs are supposed to improve children's growth. The impact of such a program was assessed in 4 municipalities in northeast Brazil by comparing 1387 children under 7 y of age from program beneficiary households with 502 matched nonbeneficiaries who were selected to receive the program but who subsequently were excluded as a result of quasi random administrative errors. Anthropometric status was assessed 6 mo after benefits began to be distributed, and beneficiary children were 0.13 Z-scores lighter (weight-for-age) than excluded children, after adjusting for confounders (P = 0.024). The children's growth trajectories were reconstructed by copying up to 10 recorded weights from their Ministry of Health growth monitoring cards and by relating each weight to the child's age, gender, and duration of receipt of the program benefit in a random effects regression model. Totals of 472 beneficiary and 158 excluded children under 3 y of age were included in this analysis. Each additional month of exposure to the program was associated with a rate of weight gain 31 g lower than that observed in excluded children of the same age (P < 0.001). This failure to respond positively to the program may have been due to a perception that benefits would be discontinued if the child started to grow well. Nutrition programs should guard against giving the impression that poor growth will be rewarded. PMID- 15333726 TI - An educational intervention to promote appropriate complementary feeding practices and physical growth in infants and young children in rural Haryana, India. AB - Complementary feeding practices are often inadequate in developing countries, resulting in a significant nutritional decline between 6 and 18 mo of age. We assessed the effectiveness of an educational intervention to promote adequate complementary feeding practices that would be feasible to sustain with existing resources. The study was a cluster randomized controlled trial in communities in the state of Haryana in India. We developed the intervention through formative research. Eight communities were pair matched on their baseline characteristics; one of each pair was randomly assigned to receive the intervention and the other to no specific feeding intervention. Health and nutrition workers in the intervention communities were trained to counsel on locally developed feeding recommendations. Newborns were enrolled in all of the communities (552 in the intervention and 473 in the control) and followed up every 3 mo to the age of 18 mo. The main outcome measures were weights and lengths at 6, 9, 12, and 18 mo and complementary feeding practices at 9 and 18 mo. All analyses were by intent to treat. In the overall analyses, there was a small but significant effect on length gain in the intervention group (difference in means 0.32 cm, 95% CI, 0.03, 0.61). The effect was greater in the subgroup of male infants (difference in mean length gain 0.51 cm, 95% CI 0.03, 0.98). Weight gain was not affected. Energy intakes from complementary foods overall were significantly higher in the intervention group children at 9 mo (mean +/- SD: 1556 +/- 1109 vs. 1025 +/- 866 kJ; P < 0.001) and 18 mo (3807 +/- 1527 vs. 2577 +/- 1058 kJ; P < 0.001). Improving complementary feeding practices through existing services is feasible but the effect on physical growth is limited. Factors that limit physical growth in such settings must be better understood to plan more effective nutrition programs. PMID- 15333727 TI - Once-weekly and 5-days a week iron supplementation differentially affect cognitive function but not school performance in Thai children. AB - Many studies have reported comparable hemoglobin response in subjects given intermittent and daily iron supplements. However, the effect of intermittent iron supplementation on impaired cognitive function, one of the serious consequences of iron deficiency among children, has not been studied. We investigated the effects of 1 d/wk (weekly) and 5 d/wk (daily) iron supplementation on changes in results of intelligence quotient (IQ), Thai language, and mathematics tests among Thai primary schoolchildren. A double-blind, randomized, placebo-controlled trial was conducted. Primary schoolchildren (n = 397) were randomly assigned to receive iron supplements daily or weekly or placebo. Ferrous sulfate (300 mg) or placebo tablets were given under direct observation by the researcher for 16 wk. Changes in IQ, and Thai language and mathematics scores were then compared. The increases in hemoglobin concentration were comparable in the weekly and daily iron supplementation groups but serum ferritin increased more in the children supplemented daily. Children receiving daily iron supplements, however, had a significantly lower increase in IQ (3 +/- 12 points) than those receiving the supplement weekly (6 +/- 12 points) or placebo (6 +/- 12 points), whereas the last-mentioned two groups did not differ. Z-scores of Thai language and mathematics test results did not differ among the groups. We conclude that weekly iron supplementation is the regimen of choice in this study community. PMID- 15333728 TI - Alkalinization of urinary pH accelerates renal excretion of ochratoxin A in pigs. AB - The mycotoxin ochratoxin A (OA) is a cause of endemic nephropathy in farm animals and humans. Reabsorption of OA along the nephron results from nonionic diffusion and by carrier-mediated mechanisms, indicating that urine alkalinization may help to accelerate OA excretion and thus reduce its toxicity. The aim of the present study was to investigate the effect of a dietary sodium bicarbonate supplementation as a means of increasing urinary pH on the systemic availability and excretion of OA in pigs. Dietary supplementation of 2% sodium bicarbonate significantly increased urinary pH (5.7 +/- 0.2 to 8.3 +/- 0.1) and daily urine volume (1108 +/- 276 to 2479 +/- 912 mL). The systemic availability of OA and its dechloro-analog, Ochratoxin B (OB), calculated as the area under the curve (AUC) was reduced to 75 and 68%, respectively, of the control group (P < 0.05). This effect was due mainly to an accelerated elimination of OA and OB in the urine. The faster renal elimination may be due to reduced reabsorption of the ochratoxins by nonionic diffusion, and other H(+)-dependent mechanisms. Thus, urinary alkalinization may be an efficient means to partially reduce the toxic effects of OA in pigs. PMID- 15333729 TI - Maternal adaptive immunity influences the intestinal microflora of suckling mice. AB - The microflorae in the intestine of breast-fed infants are distinct from those that typically populate the intestine of formula-fed infants. Although the acquisition of passive immunity through breast-feeding may play a critical role in influencing the pattern of bacterial colonization of the gut, the precise mechanisms underlying the differences in the commensal microflorae of breast and formula-fed children have not been established. We hypothesized that the assemblage of commensal microflorae in suckling and weaned mice may be influenced by the maternal adaptive immune system. To test this hypothesis, we analyzed the intestinal microflorae of mice reared in the presence (wild-type) or absence of an intact maternal immune system (T- and B-cell deficient). Several types of bacteria (Lactobacillus, Enterococcus, Clostridium perfringens, Bifidobacterium, and Bacteroides) were isolated and enumerated from both the small and large intestine of 10-, 18-, 25- and 40- to 60-d old mice using selective media. The densities of bacteria were significantly lower in the small intestine of weaned mice that were reared by wild-type (WT) compared with immunodeficient (ID) dams. However, the microflorae were generally more abundant in the large intestine of suckling pups reared by WT compared with ID dams. Our results indicate that intestinal microflorae change throughout the suckling phase of development and that the maternal adaptive immune system influences the pattern and abundance of bacteria within the gut in an age- and site-specific manner. PMID- 15333730 TI - Diets deficient in indispensable amino acids rapidly decrease the concentration of the limiting amino acid in the anterior piriform cortex of rats. AB - Diets deficient in an indispensable amino acid have long been known to suppress food intake in rats. Detection of dietary deficiency takes place in the anterior piriform cortex (APC). Recent studies showed that the response to amino acid deficiency takes as little as 15 min to develop, but few data exist to correlate the concentration of amino acids in the APC with this rapid response. The purpose of this study was to measure the concentration of amino acids in the APC in a behaviorally relevant time frame. Rats were preconditioned by consumption of a basal diet for 7-10 d, and then given a test diet with either a control or deficient amino acid profile. Both the threonine- and leucine-deficient diets reliably depleted threonine and leucine concentration in the APC within 30 min, respectively. The control diets and a diet lacking the dispensable amino acid glycine did not lead to amino acid depletion. In combination with previous studies, the present results show that the decrease in the concentration of indispensable amino acids in the APC may be the initial sensory signal for recognition of dietary amino acid deficiency. PMID- 15333731 TI - The Three-Factor Eating Questionnaire-R18 is able to distinguish among different eating patterns in a general population. AB - A revised version of the Three-Factor Eating Questionnaire (TFEQ) was developed in an obese population, but its applicability to the general population was not assessed. We aimed to define the relationship between eating behavior and reported food intake. This was a cross-sectional study of 529 middle-aged adults and 358 teenagers and young adults recruited on a geographical basis. The TFEQ R18 measures 3 aspects of eating behavior: cognitive restraint (CR), uncontrolled eating (UE), and emotional eating. Reported food intake was calculated from a food frequency questionnaire. Girls who scored higher on restrained eating had a lower energy intake than the other girls (9164 kJ vs. 13,163 kJ, P < 0.001). In adult men, energy intake increased with UE (9663 kJ vs. 11,029 kJ in the lower and higher UE tertiles, respectively, P < 0.05). When specific food groups were analyzed, higher CR was positively associated in adults with healthy food groups like green vegetables [OR = 1.92 (0.68-2.44)] and negatively associated with French fries [OR = 0.35 (0.22-0.57)] and sugar [OR = 0.38 (0.23-0.61)]. Energy dense foods, such as fat, were positively associated with UE [OR = 2.28 (1.46 3.57) for dietary fat]. Finally, emotional eaters had a higher snacking food intake. In teenagers and young adults, most associations were seen with CR. Converse to observations in adults, teenagers and young adults who exhibited a high cognitive restraint reported consumption of fewer energy-dense foods rather than more "healthy foods." The TFEQ-R18 was therefore able to distinguish among different eating patterns in our sample of a French general population. PMID- 15333732 TI - Coffee drinking is dose-dependently related to the risk of acute coronary events in middle-aged men. AB - Heavy coffee consumption has been associated with increased coronary heart disease (CHD) risk although many studies have not observed any relation. We studied the effect of coffee consumption, assessed with a 4-d food record, on the incidence of nonfatal acute myocardial infarction or coronary death in a cohort of 1971 men who were 42 to 60 y old and free of symptomatic CHD at baseline in 1984-1989. During a mean follow-up of 14 y, 269 participants experienced an acute coronary event. After adjustment for age, smoking, exercise ischemia, diabetes, income, and serum insulin concentration, the rate ratios (95% CIs) in daily nondrinkers and light (375 mL or less), moderate (reference level), and heavy (814 mL or more) drinkers were 0.84 (0.41-1.72), 1.22 (0.90-1.64), 1.00, and 1.43 (1.06-1.94). To address time dependence of the effect, the analysis was repeated for 75 CHD events that occurred during the first 5 y; the respective rate ratios were 0.42 (0.06-3.10), 2.00 (1.16-3.44), 1.00, and 2.07 (1.17-3.65). Further adjustment for serum HDL and LDL cholesterol concentration, diastolic blood pressure, maximal oxygen uptake, and waist-hip ratio slightly increased the rate ratio for heavy coffee intake. Neither the brewing method (boiling vs. filtering) nor the serum LDL cholesterol concentration had any impact on the risk estimates for coffee intake. In conclusion, heavy coffee consumption increases the short term risk of acute myocardial infarction or coronary death, independent of the brewing method or currently recognized risk factors for CHD. PMID- 15333733 TI - Correlates of serum lutein + zeaxanthin: findings from the Third National Health and Nutrition Examination Survey. AB - The determinants of blood levels of carotenoids were previously investigated in small or select samples. The relations of serum lutein + zeaxanthin to possible diet, lifestyle, and physiological determinants in 7059 participants of the Third National Health and Nutrition Examination Survey (1988-1994), > or = 40 y old, were examined. In a fully adjusted, multiple linear regression model, lower serum lutein + zeaxanthin was significantly associated with smoking, heavy drinking, being white, female, or not being physically active, having lower dietary lutein + zeaxanthin, higher fat-free mass, a higher percentage of fat mass, a higher waist-hip ratio, lower serum cholesterol, a higher white blood cell count, and high levels of C-reactive protein (P < 0.05). The model explained 24% of the variation present in serum lutein + zeaxanthin for the current sample. The correlation between dietary and serum lutein + zeaxanthin was 0.17 and increased to 0.18 after adjusting for the effects of given covariates. Each 10% increase in dietary lutein + zeaxanthin was associated with a 1% increase in serum conditional on other terms in the model. Many factors that influence the level of serum lutein + zeaxanthin remain unknown. PMID- 15333734 TI - Soy isoflavone intake lowers serum LDL cholesterol: a meta-analysis of 8 randomized controlled trials in humans. AB - Clinical trials have noted hypocholesterolemic effects of soy protein intake, but the components responsible are not known. This meta-analysis of 8 randomized controlled trials was conducted to more precisely evaluate the effects of isoflavones on blood LDL cholesterol concentration independently of soy protein level. PubMed was searched for English-language "randomized controlled trial" articles published from 1966 to 2003 that described the effects of soy protein isolate (SPI) intake with measured isoflavone levels on blood lipids in humans using the search terms "soy protein," "isoflavones," and "cholesterol." From 31 articles identified by the search, 8 articles (with 10 low vs. high isoflavone comparisons) were selected for the meta-analysis. Subjects in each comparison consumed similar dietary fat, cholesterol, and fiber; the reported body weight of subjects did not change significantly during treatment. Serum LDL cholesterol concentration in subjects who consumed SPI (mean 50 g/d) with high isoflavone content (mean intake 96 mg/d) decreased by 0.15 mmol/L (95% CI: 0.08 to 0.23 mmol/L; P < 0.0001) compared with those who consumed the same SPI level with low isoflavone content (mean intake 6 mg/d). Decreases in serum LDL cholesterol concentration in hypercholesterolemic and normocholesterolemic subjects were 0.18 mmol/L (95% CI: 0.01 to 0.35 mmol/L; P = 0.03) and 0.14 mmol/L (95% CI: 0.06, 0.23 mmol/L; P = 0.0008), respectively. With identical soy protein intake, high isoflavone intake led to significantly greater decreases in serum LDL cholesterol than low isoflavone intake, demonstrating that isoflavones have LDL cholesterol lowering effects independent of soy protein. PMID- 15333735 TI - Fractional synthesis rates of DNA and protein in rabbit skin are not correlated. AB - We developed a method for measurement of skin DNA synthesis, reflecting cell division, in conscious rabbits by infusing D-[U-(13)C(6)]glucose and L [(15)N]glycine. Cutaneous protein synthesis was simultaneously measured by infusion of L-[ring-(2)H(5)]phenylalanine. Rabbits were fitted with jugular venous and carotid arterial catheters, and were studied during the infusion of an amino acid solution (10% Travasol). The fractional synthetic rate (FSR) of DNA from the de novo nucleotide synthesis pathway, a reflection of total cell division, was 3.26 +/- 0.59%/d in whole skin and 3.08 +/- 1.86%/d in dermis (P = 0.38). The de novo base synthesis pathway accounted for 76 and 60% of the total DNA FSR in whole skin and dermis, respectively; the contribution from the base salvage pathway was 24% in whole skin and 40% in dermis. The FSR of protein in whole skin was 5.35 +/- 4.42%/d, which was greater (P < 0.05) than that in dermis (2.91 +/- 2.52%/d). The FSRs of DNA and protein were not correlated (P = 0.33), indicating that cell division and protein synthesis are likely regulated by different mechanisms. This new approach enables investigations of metabolic disorders of skin diseases and regulation of skin wound healing by distinguishing the 2 principal components of skin metabolism, which are cell division and protein synthesis. PMID- 15333737 TI - Proceedings of the Sixty-Eighth Annual Meeting of the American Society for Nutritional Sciences: Washington, DC April 17-21, 2004. PMID- 15333736 TI - Tissue fatty acid profiles can be used to quantify endogenous rumenic acid synthesis in lambs. AB - Proportions of vaccenic (trans-11 18:1) and rumenic (cis-9, trans-11 18:2) acids in mesenteric adipose, subcutaneous adipose, and longissimus muscle tissue lipids from lambs fed varying proportions of forages and concentrates were used to develop a mathematical model to predict exogenous and endogenous contributions to rumenic acid (RA) in the several tissues. From the model, we were able to estimate the proportion of absorbed RA, the proportion of vaccenic acid (VA) desaturated, the original proportion of VA in the tissue (before desaturation), and finally the proportion of RA synthesized endogenously. Estimates of endogenous RA were in the range of published data estimated by independent procedures. An independent data set of VA and RA in milk fat was used to challenge the model. Predictions were concordant with observations, although estimates of endogenous RA synthesis were lower than previous reports. Changing the amount of exogenous RA through manipulation of the diet influenced desaturation of VA inversely, so that endogenous RA synthesis was decreased when exogenous supply was increased (r = -0.80). The model should be challenged with data from human and nonruminant, as well as ruminant studies. PMID- 15333738 TI - Symposium introduction: nutrition and gene regulation. PMID- 15333739 TI - Nutritional regulation of mRNA processing. AB - Understanding how a cell adapts to dietary energy in the form of carbohydrate versus energy in the form of triacylglycerol requires knowledge of how the activity of the enzymes involved in lipogenesis is regulated. Changes in the activity of these enzymes are largely caused by changes in the rate at which their proteins are synthesized. Nutrients within the diet can signal these changes either via altering hormone concentrations or via their own unique signal transduction pathways. Most of the lipogenic genes are regulated by changes in the rate of their transcription. Glucose-6-phosphate dehydrogenase (G6PD) is unique in this group of enzymes in that nutritional regulation of its synthesis involves steps exclusively at a posttranscriptional level. G6PD activity is enhanced by the consumption of diets high in carbohydrate and is inhibited by the consumption of polyunsaturated fat. In this review, evidence is presented that changes in the rate of synthesis of the mature G6PD mRNA involves regulation of the efficiency of splicing of the nascent G6PD transcript. Furthermore, this regulation involves the activity of a cis-acting sequence in the G6PD primary transcript. This sequence in exon 12 is essential for the inhibition of G6PD mRNA splicing by PUFA. Understanding the mechanisms by which nutrients alter nuclear posttranscriptional events will provide new information on the breadth of mechanisms involved in gene regulation. PMID- 15333740 TI - Control of gene expression by fatty acids. AB - The last decade provided evidence that major (glucose, fatty acids, amino acids) or minor (iron, vitamin, etc.) dietary constituents regulated gene expression in an hormonal-independent manner. This review focuses on molecular mechanisms by which fatty acids control the expression genes encoding regulatory protein involved in their own metabolism. Nonesterified fatty acids or their CoA derivatives seem to be the main signals involved in the transcriptional effect of long-chain fatty acids. The effects of fatty acids are mediated either directly owing to their specific binding to various nuclear receptors (PPAR, LXR, HNF 4alpha) leading to changes in the trans-activating activity of these transcription factors, or indirectly as the result of changes in the abundance of regulatory transcription factors (SREBP-1c, ChREBP, etc.). The relative contribution of each transcription factor in fatty acid-induced positive or negative gene expression is discussed. PMID- 15333741 TI - Short chain fatty acid regulation of signaling genes expressed by the intestinal epithelium. AB - Changes in diet greatly affect the mucosal immune system, particularly in diseases such as Crohn's disease and necrotizing enterocolitis. This review examines the hypothesis that alterations in the luminal environment of the intestine regulate the expression of genes in the epithelium responsible for signaling to immune cells. Increasing chemokine expression in the mouse intestinal epithelium using transgenic techniques enhances the recruitment of neutrophils and lymphocytes into the intestine. Furthermore, SCFA concentrations in the intestinal lumen vary markedly with diet. SCFAs alter chemokine expression by inhibiting histone deacetylase activity in the enterocyte. The review therefore describes a molecular pathway explaining how changes in diet may alter leukocyte recruitment by regulating enterocyte gene expression. It is likely that other similar pathways remain to be discovered. PMID- 15333742 TI - Leptin and the control of metabolism: role for stearoyl-CoA desaturase-1 (SCD-1). AB - The incidence of obesity has increased sharply in recent years, making it one of the most urgent public health concerns worldwide. The hormone leptin is the central mediator in a negative feedback loop regulating energy homeostasis. Leptin administration leads to reduced food intake, increased energy expenditure, and weight loss. Leptin also mediates unique metabolic effects, specifically depleting lipid from liver and other peripheral tissues. While elucidation of leptin's role has permitted a more detailed view of the biology underlying energy homeostasis, most obese individuals are leptin resistant. A more complete understanding of the molecular components of the leptin pathway is necessary to develop effective treatment for obesity and the Metabolic Syndrome. We review here studies on the identification of one such component, stearoyl-CoA desaturase 1 (SCD-1), as a gene specifically repressed by leptin and discuss the role of this process in mediating the metabolic effects of leptin. Data indicate that pharmacologic manipulation of SCD-1 may be of benefit in the treatment of obesity, diabetes, hepatic steatosis, and other components of the Metabolic Syndrome. PMID- 15333744 TI - The role of leptin in regulating neuroendocrine function in humans. AB - Eating disorders are a group of disease states including anorexia nervosa, bulimia nervosa and binge eating on one end as well as episodic or chronic overeating resulting in obesity at the other end of the spectrum. These disorders are characterized by decreased and/or increased energy intake and are frequently associated with hormonal and metabolic disorders. The discovery of leptin, an adipocyte-secreted hormone acting in the brain to regulate energy homeostasis, and its subsequent study in human physiology have significantly advanced our understanding of normal human physiology and have provided new opportunities for understanding and possibly treating disease states, such as anorexia and bulimia nervosa. It has been recently discovered that leptin levels above a certain threshold are required to activate the hypothalamic-pituitary-gonadal and hypothalamic-pituitary-thyroid axes in men, whereas the hypothalamic-pituitary adrenal, renin-aldosterone, and growth hormone-IGF-1 axes may be largely independent of circulating leptin levels in humans. In this review, we summarize the latest findings related to the role of leptin in the regulation of several neuroendocrine axes, such as the hypothalamic-pituitary-gonadal and the hypothalamic-pituitary-thyroid axes in humans and discuss its potential pathophysiologic role in eating disorders. PMID- 15333743 TI - Fatty acid binding proteins--the evolutionary crossroads of inflammatory and metabolic responses. AB - Fatty acid binding proteins (FABPs) are members of a highly conserved family of proteins with the task of protecting a cell's delicate lipid balance. Yet they fail when faced with metabolic or inflammatory stress, turning the cytosol into an inhospitable environment with less than ideal outcomes. This review will focus on how FABPs direct lipid traffic and simultaneously control inflammatory and metabolic pathways under the pressures of the Metabolic Syndrome. PMID- 15333745 TI - Alcohol, one-carbon metabolism, and colorectal cancer: recent insights from molecular studies. AB - A growing body of evidence implicates alcohol intake as a risk factor for colorectal cancer. Until recently, most of the data were based on epidemiologic data that examined alcohol intake in relation to risk of colorectal neoplasia, but the evidence has now been broadened by recent molecular studies on mechanisms. In particular, a number of observations strongly support a role for alcohol acting through disruptions in one-carbon metabolism. Excessive alcohol intake is a fairly consistent risk factor for colorectal neoplasia in most studies, and studies show much higher risks of colorectal neoplasia in those with high alcohol and low folate than with high alcohol or low folate individually. Interactions between high alcohol-low folate and the MTHFR677TT genotype with risk of colorectal neoplasia suggest that alcohol is acting through its effects on one-carbon metabolism because the combination of high alcohol-low folate and the MTHFR677TT genotype are related to markedly elevated serum levels of homocysteine and to DNA hypomethylation. In addition, in Japanese studies, consumers of alcohol possessing the ALDH2*2 allele, who have very elevated levels of acetaldehyde, are at high risk for colorectal cancer. The relatively high prevalence of the ALDH2*2 allele may thus account for the stronger association between alcohol and colorectal neoplasia that is frequently observed in studies in Japanese populations. Further research integrating studies with more detailed data on alcohol intake levels and patterns, folate and other related nutrient status, and relevant genotypes may help clarify unresolved questions regarding the health consequences of moderate to high alcohol drinking. PMID- 15333746 TI - Diet-gene interactions in p53-deficient mice: insulin-like growth factor-1 as a mechanistic target. AB - Progress in cancer prevention research is being facilitated by the use of animal models displaying specific genetic susceptibilities for cancer, such as mice deficient in one (+/-) or both (-/-) alleles of the p53 tumor suppressor gene. Our lab, which focuses on nutrition (particularly energy balance/obesity) and molecular carcinogenesis, has shown in p53-/- mice that calorie restriction (CR) increases the latency of spontaneous tumor development (mostly lymphomas) approximately 75%, decreases serum insulin-like growth factor (IGF)-1 and leptin levels, and induces apoptosis in immature (lymphoma-susceptible) thymocytes. In heterozygous p53-deficient (p53+/-) mice, CR and a one day/wk fast each significantly delay spontaneous tumor development (a mix of lymphomas, sarcomas, and epithelial tumors) and decreases serum IGF-1 and leptin levels, even when begun late in life. We are presently comparing and combining CR and exercise (treadmill and running wheel) to further elucidate the relationships between energy balance, p53, and tumorigenesis in these models. Furthermore, we have capitalized on the susceptibility of p53+/- mice to chronic, low-dose aromatic amine-induced bladder carcinogenesis to develop a model for evaluating bladder cancer prevention approaches. Using this model, we have established that IGF-1 mediates many of the anti-cancer effects of CR. We are currently conducting oligonucleotide microarray studies to further characterize diet-gene interactions underlying the anti-cancer effects of CR and to determine which of the CR responsive genes are IGF-1 dependent. PMID- 15333747 TI - Translational regulation of gene expression by omega-3 fatty acids. AB - The incidence of some cancers shows dramatic variations around the world that cannot be explained by ethnic or racial differences. Observational studies point to a negative correlation between consumption of fish and incidence of breast and prostate cancer. In vitro and animal model studies indicate that (omega-3) PUFAs present at high concentrations in marine animals inhibit proliferation of cancer cells and growth of tumors. However, how these fatty acids inhibit cell proliferation and tumor growth is a matter of considerable debate. In this review we summarize our recent work indicating that Ca(++) depletion mediated phosphorylation of the alpha subunit of eIF2 and subsequent inhibition of translation initiation account for the anti-cancer activity of (omega-3) PUFAs. PMID- 15333748 TI - Molecular targets for bioactive food components. AB - Mounting evidence points to dietary habits as an important determinant of cancer risk and tumor behavior. Although the linkages with diet are intriguing, the literature is also laden with inconsistencies. The reasons for these inconsistencies are likely multi-factorial, but probably reflect variations in the ability of bioactive constituents to reach or affect critical molecular targets. Fluctuations in the foods consumed not only influence the intake of particular bioactive components, but may alter metabolism and potentially influence the sites of action of both essential and nonessential nutrients. Genetic polymorphisms are increasingly recognized as another factor that can alter the response to dietary components (nutritional transcriptomic effect) by influencing the absorption, metabolism, or sites of action. Likewise, variation in DNA methylation patterns and other epigenetic events that influence overall gene expression can be influenced by dietary intakes. Furthermore, variation in the ability of food components to increase or depress gene expression (nutrigenomic effect) may account for some of the observed inconsistencies in the response to dietary change. Because a host of food components are recognized to influence phosphorylation and other posttranslational events, it is also likely that these and other proteomic modifications account for at least part of the response and variation that is reported in the literature. Collectively, it is clear that bioactive food components can influence a number of key molecular events that are involved in health and disease resistance. As the era of molecular nutrition unfolds, a greater understanding of how these foods and components influence cancer will surely arise. Such information will be critical in the development of effective tailored strategies for reducing cancer burden. Just as important, however, is that as this information unfolds it is utilized within a responsible bioethical framework. PMID- 15333749 TI - Incorporation of a single His residue by rational design enables thiol-ester hydrolysis by human glutathione transferase A1-1. AB - A strategy for rational enzyme design is reported and illustrated by the engineering of a protein catalyst for thiol-ester hydrolysis. Five mutants of human glutathione (GSH; gamma-Glu-Cys-Gly) transferase A1-1 were designed in the search for a catalyst and to provide a set of proteins from which the reaction mechanism could be elucidated. The single mutant A216H catalyzed the hydrolysis of the S-benzoyl ester of GSH under turnover conditions with a k(cat)/K(M) of 156 M(-1) x min(-1), and a catalytic proficiency of >10(7) M(-1) when compared with the first-order rate constant of the uncatalyzed reaction. The wild-type enzyme did not hydrolyze the substrate, and thus, the introduction of a single histidine residue transformed the wild-type enzyme into a turnover system for thiol-ester hydrolysis. By kinetic analysis of single, double, and triple mutants, as well as from studies of reaction products, it was established that the enzyme A216H catalyzes the hydrolysis of the thiol-ester substrate by a mechanism that includes an acyl intermediate at the side chain of Y9. Kinetic measurements and the crystal structure of the A216H GSH complex provided compelling evidence that H216 acts as a general-base catalyst. The introduction of a single His residue into human GSH transferase A1-1 created an unprecedented enzymatic function, suggesting a strategy that may be of broad applicability in the design of new enzymes. The protein catalyst has the hallmarks of a native enzyme and is expected to catalyze various hydrolytic, as well as transesterification, reactions. PMID- 15333750 TI - Gene-based vaccines and immunotherapeutics. AB - DNA vaccines, comprised of plasmid DNA encoding proteins from pathogens, allergens, and tumors, are being evaluated as prophylactic vaccines and therapeutic treatments for infectious diseases, allergies, and cancer; plasmids encoding normal human proteins are likewise being tested as vaccines and treatments for autoimmune diseases. Examples of in vivo prophylaxis and immunotherapy, based on different types of immune responses (humoral and cellular), in a variety of disease models and under evaluation in early phase human clinical trials are presented. Viral vectors continue to show better levels of expression than those achieved by DNA plasmid vectors. We have focused our clinical efforts, at this time, on the use of recombinant viral vectors for both vaccine as well as cytokine gene transfer studies. We currently have four clinical programs in cancer immunotherapy. Two nonspecific immunotherapy programs are underway that apply adenoviral vectors for the transfer of cytokine genes into tumors in situ. An adenovirus-IFN gamma construct (TG1042) is currently being tested in phase II clinical trials in cutaneous lymphoma. A similar construct, adenovirus-IL2 (TG1024), also injected directly into solid tumors, is currently being tested in patients with solid tumors (about one-half of which are melanoma). Encouraging results are seen in both programs. Two cancer vaccine immunotherapy programs focus on two cancer-associated antigens: human papilloma virus E6 and E7 proteins and the epithelial cancer-associated antigen MUC1. Both are encoded by a highly attenuated vaccinia virus vector [modified vaccinia Ankara (MVA)] and both are coexpressed with IL-2. Encouraging results seen in both of these programs are described. PMID- 15333751 TI - Enhanced photosynthesis and redox energy production contribute to salinity tolerance in Dunaliella as revealed by homology-based proteomics. AB - Salinity is a major limiting factor for the proliferation of plants and inhibits central metabolic activities such as photosynthesis. The halotolerant green alga Dunaliella can adapt to hypersaline environments and is considered a model photosynthetic organism for salinity tolerance. To clarify the molecular basis for salinity tolerance, a proteomic approach has been applied for identification of salt-induced proteins in Dunaliella. Seventy-six salt-induced proteins were selected from two-dimensional gel separations of different subcellular fractions and analyzed by mass spectrometry (MS). Application of nanoelectrospray mass spectrometry, combined with sequence-similarity database-searching algorithms, MS BLAST and MultiTag, enabled identification of 80% of the salt-induced proteins. Salinity stress up-regulated key enzymes in the Calvin cycle, starch mobilization, and redox energy production; regulatory factors in protein biosynthesis and degradation; and a homolog of a bacterial Na(+)-redox transporters. The results indicate that Dunaliella responds to high salinity by enhancement of photosynthetic CO(2) assimilation and by diversion of carbon and energy resources for synthesis of glycerol, the osmotic element in Dunaliella. The ability of Dunaliella to enhance photosynthetic activity at high salinity is remarkable because, in most plants and cyanobacteria, salt stress inhibits photosynthesis. The results demonstrated the power of MS BLAST searches for the identification of proteins in organisms whose genomes are not known and paved the way for dissecting molecular mechanisms of salinity tolerance in algae and higher plants. PMID- 15333752 TI - A complementary bioinformatics approach to identify potential plant cell wall glycosyltransferase-encoding genes. AB - Plant cell wall (CW) synthesizing enzymes can be divided into the glycan (i.e. cellulose and callose) synthases, which are multimembrane spanning proteins located at the plasma membrane, and the glycosyltransferases (GTs), which are Golgi localized single membrane spanning proteins, believed to participate in the synthesis of hemicellulose, pectin, mannans, and various glycoproteins. At the Carbohydrate-Active enZYmes (CAZy) database where e.g. glucoside hydrolases and GTs are classified into gene families primarily based on amino acid sequence similarities, 415 Arabidopsis GTs have been classified. Although much is known with regard to composition and fine structures of the plant CW, only a handful of CW biosynthetic GT genes-all classified in the CAZy system-have been characterized. In an effort to identify CW GTs that have not yet been classified in the CAZy database, a simple bioinformatics approach was adopted. First, the entire Arabidopsis proteome was run through the Transmembrane Hidden Markov Model 2.0 server and proteins containing one or, more rarely, two transmembrane domains within the N-terminal 150 amino acids were collected. Second, these sequences were submitted to the SUPERFAMILY prediction server, and sequences that were predicted to belong to the superfamilies NDP-sugartransferase, UDP glycosyltransferase/glucogen-phosphorylase, carbohydrate-binding domain, Gal binding domain, or Rossman fold were collected, yielding a total of 191 sequences. Fifty-two accessions already classified in CAZy were discarded. The resulting 139 sequences were then analyzed using the Three-Dimensional-Position Specific Scoring Matrix and mGenTHREADER servers, and 27 sequences with similarity to either the GT-A or the GT-B fold were obtained. Proof of concept of the present approach has to some extent been provided by our recent demonstration that two members of this pool of 27 non-CAZy-classified putative GTs are xylosyltransferases involved in synthesis of pectin rhamnogalacturonan II (J. Egelund, B.L. Petersen, A. Faik, M.S. Motawia, C.E. Olsen, T. Ishii, H. Clausen, P. Ulvskov, and N. Geshi, unpublished data). PMID- 15333753 TI - AraPerox. A database of putative Arabidopsis proteins from plant peroxisomes. AB - To identify unknown proteins from plant peroxisomes, the Arabidopsis genome was screened for proteins with putative major or minor peroxisome targeting signals type 1 or 2 (PTS1 or PTS2), as defined previously (Reumann S [2004] Plant Physiol 135: 783-800). About 220 and 60 proteins were identified that carry a putative PTS1 or PTS2, respectively. To further support postulated targeting to peroxisomes, several prediction programs were applied and the putative targeting domains analyzed for properties conserved in peroxisomal proteins and for PTS conservation in homologous plant expressed sequence tags. The majority of proteins with a major PTS and medium to high overall probability of peroxisomal targeting represent novel nonhypothetical proteins and include several enzymes involved in beta-oxidation of unsaturated fatty acids and branched amino acids, and 2-hydroxy acid oxidases with a predicted function in fatty acid alpha oxidation, as well as NADP-dependent dehydrogenases and reductases. In addition, large protein families with many putative peroxisomal isoforms were recognized, including acyl-activating enzymes, GDSL lipases, and small thioesterases. Several proteins are homologous to prokaryotic enzymes of a novel aerobic hybrid degradation pathway for aromatic compounds and proposed to be involved in peroxisomal biosynthesis of plant hormones like jasmonic acid, auxin, and salicylic acid. Putative regulatory proteins of plant peroxisomes include protein kinases, small heat shock proteins, and proteases. The information on subcellular targeting prediction, homology, and in silico expression analysis for these Arabidopsis proteins has been compiled in the public database AraPerox to accelerate discovery and experimental investigation of novel metabolic and regulatory pathways of plant peroxisomes. PMID- 15333754 TI - Genomic analysis of the nitrate response using a nitrate reductase-null mutant of Arabidopsis. AB - A nitrate reductase (NR)-null mutant of Arabidopsis was constructed that had a deletion of the major NR gene NIA2 and an insertion in the NIA1 NR gene. This mutant had no detectable NR activity and could not use nitrate as the sole nitrogen source. Starch mobilization was not induced by nitrate in this mutant but was induced by ammonium, indicating that nitrate was not the signal for this process. Microarray analysis of gene expression revealed that 595 genes responded to nitrate (5 mm nitrate for 2 h) in both wild-type and mutant plants. This group of genes was overrepresented most significantly in the functional categories of energy, metabolism, and glycolysis and gluconeogenesis. Because the nitrate response of these genes was NR independent, nitrate and not a downstream metabolite served as the signal. The microarray analysis also revealed that shoots can be as responsive to nitrate as roots, yet there was substantial organ specificity to the nitrate response. PMID- 15333755 TI - Arabidopsis Cys2/His2-type zinc-finger proteins function as transcription repressors under drought, cold, and high-salinity stress conditions. AB - ZPT2-related proteins that have two canonical Cys-2/His-2-type zinc-finger motifs in their molecules are members of a family of plant transcription factors. To characterize the role of this type of protein, we analyzed the function of Arabidopsis L. Heynh. genes encoding four different ZPT2-related proteins (AZF1, AZF2, AZF3, and STZ). Gel-shift analysis showed that the AZFs and STZ bind to A(G/C)T repeats within an EP2 sequence, known as a target sequence of some petunia (Petunia hybrida) ZPT2 proteins. Transient expression analysis using synthetic green fluorescent protein fusion genes indicated that the AZFs and STZ are preferentially localized to the nucleus. These four ZPT2-related proteins were shown to act as transcriptional repressors that down-regulate the transactivation activity of other transcription factors. RNA gel-blot analysis showed that expression of AZF2 and STZ was strongly induced by dehydration, high salt and cold stresses, and abscisic acid treatment. Histochemical analysis of beta-glucuronidase activities driven by the AZF2 or STZ promoters revealed that both genes are induced in leaves rather than roots of rosette plants by the stresses. Transgenic Arabidopsis overexpressing STZ showed growth retardation and tolerance to drought stress. These results suggest that AZF2 and STZ function as transcriptional repressors to increase stress tolerance following growth retardation. PMID- 15333756 TI - Light regulation of the Arabidopsis respiratory chain. Multiple discrete photoreceptor responses contribute to induction of type II NAD(P)H dehydrogenase genes. AB - Controlled oxidation reactions catalyzed by the large, proton-pumping complexes of the respiratory chain generate an electrochemical gradient across the mitochondrial inner membrane that is harnessed for ATP production. However, several alternative respiratory pathways in plants allow the maintenance of substrate oxidation while minimizing the production of ATP. We have investigated the role of light in the regulation of these energy-dissipating pathways by transcriptional profiling of the alternative oxidase, uncoupling protein, and type II NAD(P)H dehydrogenase gene families in etiolated Arabidopsis seedlings. Expression of the nda1 and ndc1 NAD(P)H dehydrogenase genes was rapidly up regulated by a broad range of light intensities and qualities. For both genes, light induction appears to be a direct transcriptional effect that is independent of carbon status. Mutant analyses demonstrated the involvement of two separate photoreceptor families in nda1 and ndc1 light regulation: the phytochromes (phyA and phyB) and an undetermined blue light photoreceptor. In the case of the nda1 gene, the different photoreceptor systems generate distinct kinetic induction profiles that are integrated in white light response. Primary transcriptional control of light response was localized to a 99-bp region of the nda1 promoter, which contains an I-box flanked by two GT-1 elements, an arrangement prevalent in the promoters of photosynthesis-associated genes. Light induction was specific to nda1 and ndc1. The only other substantial light effect observed was a decrease in aox2 expression. Overall, these results suggest that light directly influences the respiratory electron transport chain via photoreceptor-mediated transcriptional control, likely for supporting photosynthetic metabolism. PMID- 15333757 TI - Seed-specific heterologous expression of a nasturtium FAE gene in Arabidopsis results in a dramatic increase in the proportion of erucic acid. AB - The fatty acid elongase [often designated FAE or beta-(or 3-) ketoacyl-CoA synthase] is a condensing enzyme and is the first component of the elongation complex involved in synthesis of erucic acid (22:1) in seeds of garden nasturtium (Tropaeolum majus). Using a degenerate primers approach, a cDNA of a putative embryo FAE was obtained showing high homology to known plant elongases. This cDNA contains a 1,512-bp open reading frame that encodes a protein of 504 amino acids. A genomic clone of the nasturtium FAE was isolated and sequence analyses indicated the absence of introns. Northern hybridization showed the expression of this nasturtium FAE gene to be restricted to the embryo. Southern hybridization revealed the nasturtium beta-ketoacyl-CoA synthase to be encoded by a small multigene family. To establish the function of the elongase homolog, the cDNA was introduced into two different heterologous chromosomal backgrounds (Arabidopsis and tobacco [Nicotiana tabacum]) under the control of a seed-specific (napin) promoter and the tandem 35S promoter, respectively. Seed-specific expression resulted in up to an 8-fold increase in erucic acid proportions in Arabidopsis seed oil, while constitutive expression in transgenic tobacco tissue resulted in increased proportions of very long chain saturated fatty acids. These results indicate that the nasturtium FAE gene encodes a condensing enzyme involved in the biosynthesis of very long chain fatty acids, utilizing monounsaturated and saturated acyl substrates. Given its strong and unique preference for elongating 20:1-CoA, the utility of the FAE gene product for directing or engineering increased synthesis of erucic acid is discussed. PMID- 15333758 TI - Embryo-specific reduction of ADP-Glc pyrophosphorylase leads to an inhibition of starch synthesis and a delay in oil accumulation in developing seeds of oilseed rape. AB - In oil-storing Brassica napus (rape) seeds, starch deposition occurs only transiently in the early stages of development, and starch is absent from mature seeds. This work investigates the influence of a reduction of ADP-Glc pyrophosphorylase (AGPase) on storage metabolism in these seeds. To manipulate the activity of AGPase in a seed-specific manner, a cDNA encoding the small subunit of AGPase was expressed in the sense or antisense orientation under the control of an embryo-specific thioesterase promoter. Lines were selected showing an embryo-specific decrease in AGPase due to antisense and cosuppression at different stages of development. At early developmental stages (25 days after flowering), a 50% decrease in AGPase activity was accompanied by similar decreases in starch content and the rate of starch synthesis measured by injecting (14)C-Suc into seeds in planta. In parallel to inhibition of starch synthesis, the level of ADP-Glc decreased, whereas Glc 1-phosphate levels increased, providing biochemical evidence that inhibition of starch synthesis was due to repression of AGPase. At 25 days after flowering, repression of starch synthesis also led to a decrease in the rate of (14)C-Suc degradation and its further metabolism via other metabolic pathways. This was not accompanied by an increase in the levels of soluble sugars, indicating that Suc import was inhibited in parallel. Flux through glycolysis, the activities of hexokinase, and inorganic pyrophosphate-dependent phosphofructokinase, and the adenylate energy state (ATP to ADP ratio) of the transgenic seeds decreased, indicating inhibition of glycolysis and respiration compared to wild type. This was accompanied by a marked decrease in the rate of storage lipid (triacylglycerol) synthesis and in the fatty acid content of seeds. In mature seeds, glycolytic enzyme activities, metabolite levels, and ATP levels remained unchanged, and the fatty acid content was only marginally lower compared to wild type, indicating that the influence of AGPase on carbon metabolism and oil accumulation was largely compensated for in the later stages of seed development. Results indicate that AGPase exerts high control over starch synthesis at early stages of seed development where it is involved in establishing the sink activity of the embryo and the onset of oil accumulation. PMID- 15333759 TI - Intrinsic cardiac catecholamines help maintain beating activity in neonatal rat cardiomyocyte cultures. AB - In the present study, we identify intrinsic cardiac adrenergic (ICA) cells in the neonatal rat heart using immunofluorescent histochemical staining techniques with antibodies that specifically recognize the major enzymes in the catecholamine biosynthetic pathway. ICA cells are most concentrated near the endocardial surface of ventricular myocardium, but are also found sporadically throughout the heart. In primary cultures of neonatal rat cardiomyocytes, ICA cells are closely associated with clusters of cardiomyocytes. To investigate a potential role for intrinsically produced catecholamines, we recorded beating rates in the presence and absence of the catecholamine-depleting agent reserpine or the adrenergic receptor blockers prazosin and timolol using videomicroscopy and photodiode sensors. Our results show that beating rates slow significantly when endogenous catecholamines are depleted or when their action is blocked with either a beta- or an alpha1-adrenergic receptor antagonist. These data indicate that intrinsic cardiac catecholamines help to maintain beating rates in neonatal rat cardiomyocyte cultures via stimulation of alpha1- and beta-adrenergic receptors. This information should help to increase our understanding of the physiologic mechanisms governing cardiovascular function in neonates. PMID- 15333760 TI - A chicken model for DGCR6 as a modifier gene in the DiGeorge critical region. AB - DGCR6 is the most centromeric gene in the human DiGeorge critical region and is the only gene in the region with a second functional copy on a repeat localized more distally on chromosome 22. We isolated the chicken ortholog of DGCR6 and showed an embryonic expression pattern that is initially broad but becomes gradually restricted to neural crest cell derivatives of the cardiovasculature. Retrovirus based gene transduction was used to deliver sense and antisense messages to premigrating neural crest cells in vivo. Embryos in which DGCR6 expression was attenuated revealed cardiovascular anomalies reminiscent of those found in DiGeorge syndrome. Moreover, the expression profiles of three other genes from the DiGeorge critical region, TBX-1, UFD1L, and HIRA, were shown to be altered in this model. TBX-1 and UFD1L levels were increased, whereas HIRA was decreased in the hearts and pharyngeal arches of embryos treated with antisense or partial sense constructs, but not with sense constructs for DGCR6. The expression changes were transient and followed the normal DGCR6 expression profile. These data show that neural crest cells might have a role in the distribution of modulator signals to the heart and pharyngeal arches. Moreover, it shows a repressor function for DGCR6 on the expression of TBX-1 and UFD1L. For the first time, DiGeorge syndrome is shown to be a contiguous gene syndrome in which not only several genes from the critical region, but also different cell types within the embryo, interact in the development of the phenotype. PMID- 15333762 TI - Leo G. Rigler, MD: a personal perspective. PMID- 15333763 TI - The Rigler sign and Leo G. Rigler. PMID- 15333764 TI - Stereotactic core breast biopsy of malignant calcifications: diagnostic yield of cores with and cores without calcifications on specimen radiographs. AB - PURPOSE: To retrospectively compare core biopsy diagnosis with final diagnosis at surgical excision in cores with and cores without calcification on specimen radiographs. MATERIALS AND METHODS: One hundred thirteen consecutive patients underwent vacuum-assisted 11- or 14-gauge needle stereotactic core biopsy for calcifications with malignant histologic results in core samples from 116 lesions. For each lesion, calcification was identified in at least one core at specimen radiography. Cores with and those without calcification seen on magnified specimen radiographs were separately submitted to and reported on by pathologists, who obtained additional levels in cores with calcification. All patients underwent surgical excision of the lesion area within 7 weeks. The pathologic diagnosis in core samples with and those without calcification on specimen radiographs was compared with final diagnosis at surgical excision. Fisher exact test was used for all chi(2) determinations of statistical significance. RESULTS: Cores with calcification on specimen radiographs were more likely to enable a final diagnosis of malignancy than were cores without calcification (98 [84%] vs 82 [71%] of 116; P =.02). Cores without calcification were significantly more likely to cause a diagnosis of cancer to be missed than were those with calcification on specimen radiographs (13 [11%] vs one [1%] of 116; P <.001). Underestimates of malignancy were more frequent in 14- than in 11 gauge specimens (11 [18%] of 60 vs six [10%] of 56; P =.30). Regardless of needle size, there was no significant difference in underestimation of malignancy between cores with and without radiographically evident calcification (17 [15%] vs 21 [18%] of 116; P =.60). CONCLUSION: Specimen radiography is essential to document calcification retrieval. Cores without radiographically demonstrated calcification may fail to show a malignant lesion. Separate identification of calcium-containing cores may assist the pathologist, who can more thoroughly evaluate these cores with additional levels of section. PMID- 15333765 TI - Coronary artery calcification: effect of size of field of view on multi-detector row CT measurements. AB - The effect of the size of the field of view (FOV) on coronary artery calcium measurements at multi-detector row computed tomography (CT) was assessed. Coronary multi-detector row CT was performed with an identical protocol in 100 consecutive subjects. CT images were reconstructed at different FOV sizes (210, 260, and 310 mm). Calcified coronary lesions were detected in all three image sets in 52 subjects. The FOV sizes tested for multi-detector row CT coronary screening had a negligible effect on coronary artery calcium measurements (P >/=.06). However, risk stratification decreased by one level in seven of 52 subjects when the FOV increased from 210 or 260 to 310 mm. PMID- 15333766 TI - Differentiation of extrahepatic bile duct cholangiocarcinoma from benign stricture: findings at MRCP versus ERCP. AB - PURPOSE: To retrospectively evaluate criteria for differentiating extrahepatic bile duct cholangiocarcinoma from benign cause of stricture at magnetic resonance cholangiopancreatography (MRCP) and to compare diagnostic accuracy with this modality versus endoscopic retrograde cholangiopancreatography (ERCP). MATERIALS AND METHODS: MRCP and ERCP images in 50 patients (27 with cholangiocarcinoma [18 men, nine women; mean age, 58 years] and 23 with benign cause of stricture [13 men, 10 women; mean age, 60 years]) were retrospectively reviewed to assess the appearance of bile duct strictures. Final diagnosis was based on surgical or biopsy findings. Strictures were described according to their imaging appearance (irregular or smooth margins, asymmetric or symmetric narrowing, abrupt narrowing or gradual tapering, and presence or absence of double-duct sign). Sensitivity, specificity, and accuracy of MRCP and ERCP were calculated by using ratings of confidence in image-based diagnosis. Lengths of stricture were electronically measured and compared by using the Student t test. RESULTS: Among cholangiographic criteria for malignant biliary stricture, irregular margins and asymmetric narrowing were more common in cholangiocarcinomas (24 [89%] of 27 patients) than in benign strictures (six [26%] and eight [35%] of 23 patients, respectively). Sensitivity, specificity, and accuracy of the two methods for differentiation of malignant from benign causes of biliary stricture were 81% (22 of 27), 70% (16 of 23), and 76% (38 of 50), respectively, for MRCP and 74% (20 of 27), 70% (16 of 23), and 72% (36 of 50), respectively, for ERCP. Mean length (+/- standard deviation) of cholangiocarcinomas was 30.0 mm +/- 8.5, and that of benign strictures was 13.6 mm +/- 9.1 (P <.001). CONCLUSION: Accuracy of MRCP is comparable with that of ERCP. Regardless of modality, a lengthy segment of extrahepatic bile duct stricture with irregular margin and asymmetric narrowing suggests cholangiocarcinoma, and a short segment with regular margin and symmetric narrowing suggests benign cause. PMID- 15333767 TI - Detection of synovial macrophages in an experimental rabbit model of antigen induced arthritis: ultrasmall superparamagnetic iron oxide-enhanced MR imaging. AB - PURPOSE: To evaluate intravenously administered ultrasmall superparamagnetic iron oxide (USPIO) as a marker of macrophage activity in an experimental rabbit model of antigen-induced arthritis. MATERIALS AND METHODS: Unilateral arthritis was induced by means of intraarticular injection of methylated bovine serum albumin in 10 knees of 10 rabbits that had been presensitized to the same antigen. The contralateral knees in these rabbits, as well as six knees in three other rabbits, served as controls. After onset of arthritis, all knees were imaged prior to and 24 hours after administration of USPIO. The magnetic resonance (MR) imaging protocol included T1-weighted spin-echo, T2-weighted fast spin-echo, T2* weighted gradient-echo, and short inversion time inversion-recovery sequences. Images were analyzed quantitatively and qualitatively with regard to signal characteristics and pattern. MR findings were correlated with histopathologic findings. Wilcoxon signed rank test was used to compare results of signal-to noise ratio calculations before and after USPIO administration. RESULTS: All knees with intraarticular injection of antigen suspension developed unilateral arthritis, whereas no signs of arthritis occurred in the control knees. On USPIO enhanced images obtained 24 hours after contrast agent administration, significant T1 (P =.03) and more predominantly T2* (P =.02) and T2 effects (P =.01) were evident in the synovium of all 10 arthritic knees, which reflected USPIO uptake by macrophages in the synovial tissue. To a lesser extent, T2* effects were present also within the joint effusion (P =.01). No significant changes in signal characteristics were detected in the 10 nonarthritic knees in the antigen-injected group or the six knees in the control group (P =.06-.91). Histologic examination confirmed uptake of iron in the macrophages of arthritic knees. Changes in MR signal characteristics within the arthritic synovium and synovial effusion were visually detectable after intravenous administration of USPIO. CONCLUSION: MR imaging at 1.5 T can depict USPIO uptake in phagocytic active macrophages in an antigen-induced arthritis animal model. PMID- 15333768 TI - Percutaneous nephrostomy of nondilated renal collecting systems with fluoroscopic guidance: technique and results. AB - PURPOSE: To retrospectively review experience with a double-puncture technique for percutaneous nephrostomy of nondilated renal collecting systems. MATERIALS AND METHODS: Over a 5-year period, 15 patients (26 kidneys) without calyceal dilatation at ultrasonography (US) who required nephrostomy drainage were studied. Eleven patients (21 kidneys) had ureteral leaks or fistulas, one patient (one kidney) had a renal pelvic stone, one patient (one kidney) was suspected of having a ureteral tumor, and the final two patients (three kidneys) had acute nondilated renal failure. Mean age was 62 years (range, 20-78 years); 13 patients were men and two were women. A double-puncture technique was used with fluoroscopic guidance (supplemented with US in some patients who had renal failure). After intravenous administration of iodinated contrast material, a 22 gauge needle was inserted into the opacified renal pelvis, and double-contrast pyelography was performed by using air or carbon dioxide to allow visualization and distention of the nondependent calyces for definitive renal access with an 18 gauge 5-F sheath-needle set and a hydrophilic wire. After serial dilation, a nephrostomy or nephroureteral catheter was inserted. Success and major complication rates were studied by reviewing radiologic and clinical case notes. RESULTS: Catheter placement was successful in 25 (96%) of 26 kidneys after one, two, or three passes with the sheath-needle set. In all successful cases, the calyx was accurately punctured. There were two major complications. One patient developed hematuria that required transfusion but no further intervention; another sustained a renal pelvic injury, but this was believed to be due to excessive dilation of the pelviureteral junction, not faulty renal puncture. There were no cases of septicemia. CONCLUSION: With a double-puncture technique and air or carbon dioxide distention, nephrostomy was achieved in 25 (96%) of all nondilated renal collecting systems. There were two major complications (two [8%] of 25 kidneys, two [13%] of 15 patients), but only one was secondary to the renal puncture. PMID- 15333769 TI - Asbestos-related pleuropulmonary diseases: evaluation with low-dose four-detector row spiral CT. AB - PURPOSE: To evaluate the depiction of lung and pleural asbestos-related lesions with low-dose four-detector row spiral computed tomography (CT). MATERIALS AND METHODS: Eighty-three male workers with a mean duration of occupational exposure to asbestos of 18 years underwent CT as part of a medicolegal investigation. CT examination included low-dose multi-detector row spiral CT of the entire thorax, with reconstruction of contiguous 5-mm-thick images, and thin-section CT, which served as the reference standard for the detection of pleural and parenchymal asbestos-related abnormalities. Two main groups of abnormalities were identified: (a) pleural plaques and diffuse pleural thickening and (b) thickened interstitial short lines, curvilinear subpleural lines, ground-glass opacity with or without bronchiectasis, and honeycombing. The frequencies of the depiction of these abnormalities on the low-dose multi-detector row images and the thin-section images were compared by using the McNemar test. RESULTS: No significant differences were observed between the low-dose and thin-section CT images in the depiction of either (a) parietal pleural fibrosis consisting of pleural plaques (identified in 67 [81%] vs 65 [78%] workers, P =.157), which appeared mainly as thick, calcified pleural linear structures; or (b) features of parenchymal fibrosis, which consisted of various combinations of intralobular and septal lines (identified in 12 [14%] vs 13 [16%] workers, P =.564), subpleural curvilinear lines (identified in 10 [12%] vs eight [10%] workers, P =.157), and ground-glass opacity with (identified in six [7%] vs six [7%] workers) or without (identified in five [6%] vs three [4%] workers, P =.317) traction bronchiectasis. A honeycombing pattern was depicted on only the thin-section CT images (P <.001). Emphysema (identified in 26 [31%] vs 14 [17%] workers at low-dose and thin section CT, respectively; P <.001) and noncalcified nodules (identified in 18 [22%] workers vs one [1%] worker, P <.001) were depicted significantly more frequently on the low-dose images than on the thin-section images. CONCLUSION: Low-dose multi-detector row spiral CT accurately depicts asbestos-related disease. PMID- 15333771 TI - Accelerated progression of coronary calcification: four-year follow-up in patients with stable coronary artery disease. AB - PURPOSE: To prospectively assess the 4-year progression rate of coronary artery calcium (CAC) in patients with clinically stable coronary artery disease (CAD) with multi-detector row computed tomography (CT). MATERIALS AND METHODS: The study group consisted of 382 consecutive patients. All underwent baseline dual sector spiral CT, and CT was repeated at 2 and 4 years later. Progression of CAC was assessed with measurement of the increase in total calcium score (TCS) and with repeated-measures analysis and multivariate linear regression models. Logistic regression model was used to predict incidence of new lesions. RESULTS: Eighty-seven percent (333 of 382) of the study group were men, with mean age of 65 years +/- 11, and 13% (49 of 382) were women, with mean age of 68 years +/- 11. The average TCS increased after 4 years by sixfold from baseline in the 1st quartile, and by four-, two- and 1.5-fold in the 2nd, 3rd, and 4th quartiles of baseline TCS (P <.01), respectively. Multiple linear regression analysis included age; sex; natural logarithm of baseline TCS; history of hypertension, diabetes mellitus, current smoking, hypercholesterolemia, and lipid-lowering therapy with cholesterol synthesis enzyme inhibitor (statin); and family history of premature CAD. Results demonstrated that natural logarithm of baseline TCS and history of current smoking were independent predictors of the 4th-year natural logarithm of TCS levels (R(2) = 0.85, P <.001). New lesions were diagnosed in 56 (15%) patients. History of statin therapy (odds ratio = 0.35; 95% confidence interval [CI]: 0.16, 0.77; P <.01), age with an increment of 5 years (odds ratio = 0.76; 95% CI: 0.64, 0.90; P =.01), and natural logarithm of baseline TCS (odds ratio = 0.73; 95% CI: 0.62, 0.86; P <.01) were independent predictors for new calcific lesions during 4 years. CONCLUSION: Accelerated progression of CAC during 4 years was found in clinically stable patients with CAD. PMID- 15333770 TI - Imaging of carotid arteries in symptomatic patients: cost-effectiveness of diagnostic strategies. AB - PURPOSE: To assess the cost-effectiveness of noninvasive imaging strategies in patients who have had a transient ischemic attack (TIA) or minor stroke and are suspected of having significant carotid artery stenosis. MATERIALS AND METHODS: From 1997 through 2000, 350 patients were included in a multicenter blinded consecutive cohort study. The sensitivities and specificities of duplex ultrasonography (US), magnetic resonance (MR) angiography, and these two examinations combined were estimated by using digital subtraction angiography (DSA) as the reference standard. The actual costs (from a societal perspective) of performing imaging and endarterectomy were estimated. The survival, quality of life, and costs associated with stroke were based on data reported in the literature. Markov modeling was used to predict long-term outcomes. Subsequently, a decision model was used to calculate costs, quality-adjusted life-years (QALYs), and incremental costs per QALY gained for 62 examination-treatment strategies. Extensive sensitivity analyses were performed. RESULTS: Duplex US had 88% sensitivity and 76% specificity with use of conventional cutoff criteria. MR angiography had comparable values: 92% sensitivity and 76% specificity. Combined concordant duplex US and MR angiography had superior diagnostic performance: 96% sensitivity and 80% specificity. Duplex US alone was the most efficient strategy. Adding MR angiography led to a marginal increase in QALYs gained but at prohibitive costs (cost-effectiveness ratio > 1 500 000 per QALY gained). Performing DSA owing to discordant duplex US and MR angiographic findings and to confirm duplex US and MR angiographic findings led to extra costs and QALY loss owing to complications. Sensitivity analyses revealed that duplex US as a stand alone examination remained the preferred strategy while estimates and assumptions were varied across plausible ranges. CONCLUSION: Duplex US performed without additional imaging is cost-effective in the selection of symptomatic patients suitable for endarterectomy. Adding MR angiography increases effectiveness slightly at disproportionately high costs, whereas DSA is inferior because of associated complications. PMID- 15333772 TI - Imaging of periosteal osteosarcoma: radiologic-pathologic comparison. AB - PURPOSE: To review the imaging appearance of periosteal osteosarcoma, with pathologic comparison. MATERIALS AND METHODS: Data for 40 pathologically confirmed periosteal osteosarcomas were retrospectively reviewed. Patient demographic data were recorded, and radiographs (n = 40), bone scintigrams (n = 10), angiograms (n = 2), and computed tomographic (CT) (n = 11) and magnetic resonance (MR) (n = 12) images were evaluated for lesion location and size, cortical changes, marrow involvement, and intrinsic characteristics by two musculoskeletal radiologists, with agreement by consensus. Pathology reports were reviewed for presence and predominance of histologic components (fibrous, chondroid, and osteoid), tumor grade, and marrow involvement. RESULTS: There were 25 male (62%) and 15 female (38%) patients with an age range of 10-37 years (average age, 20 years). The most frequent lesion locations were the diaphysis of the tibia (16 patients) or of the femur (15 patients). Radiographs showed a broad based soft-tissue mass that was attached to the cortex (all patients) and showed cortical thickening (33 patients), cortical scalloping/erosion (37 patients), and/or perpendicular periosteal reaction (38 patients) extending into the soft tissue mass. Soft-tissue masses were well defined in 91%-100% of cases and surrounded a median of 50%-55% of the cortex. Lesions commonly showed low attenuation at CT (10 patients) and high signal intensity on T2-weighted MR images (10 patients), reflecting the high water content of these largely chondroblastic lesions. Focal areas of adjacent marrow replacement were common at MR imaging (nine patients) but represented reactive changes unless they were in direct continuity with the overlying soft-tissue mass (this was rare, occurring in only one patient, and represented marrow invasion). Review of pathology reports revealed that all lesions contained chondroid tissue, which predominated in 34 patients. CONCLUSION: The radiologic appearance of periosteal osteosarcoma is a broad-based surface soft-tissue mass causing extrinsic erosion of thickened underlying diaphyseal cortex and perpendicular periosteal reaction extending into the soft-tissue component. Reactive marrow changes are commonly seen at MR imaging, but true marrow invasion is rare. PMID- 15333773 TI - Increasing the synthetic performance of penicillin acylase PAS2 by structure inspired semi-random mutagenesis. AB - A semi-random mutagenesis approach was followed to increase the performance of penicillin acylase PAS2 in the kinetically controlled synthesis of ampicillin from 6-aminopenicillanic acid (6-APA) and activated D-phenylglycine derivatives. We directed changes in amino acid residues to positions close to the active site that are expected to affect the catalytic performance of penicillin acylase: alpha R160, alpha F161 and beta F24. From the resulting triple mutant gene bank, six improved PAS2 mutants were recovered by screening only 700 active mutants with an HPLC-based screening method. A detailed kinetic analysis of the three most promising mutants, T23, TM33 and TM38, is presented. These mutants allowed the accumulation of ampicillin at 4-5 times higher concentrations than the wild type enzyme, using D-phenylglycine methyl ester as the acyl donor. At the same time, the loss of activated acyl donor due to the competitive hydrolytic side reactions could be reduced to <20% with the mutant enzymes compared >80% wild type PAS2. Although catalytic activity dropped by a factor of 5-10, the enhanced synthetic performance of the recovered penicillin acylase variants makes them interesting biocatalysts for the production of beta-lactam antibiotics. PMID- 15333774 TI - Expression of the C-terminus of HIV-1 reverse transcriptase p66 and p51 subunits as a single polypeptide with RNase H activity. AB - The C-terminus of the HIV-1 reverse transcriptase heterodimer was reconstructed into a single polypeptide. The construct encodes the p51 thumb (T) and connection (C) subdomains joined through a linker region to the p66 connection (C) and RNase H (R) domain. The TCCR protein was purified from insoluble fractions of Escherichia coli lysates. The TCCR construct maintains Mn(2+)-dependent RNase H activity and specifically cleaves the substrate mimicking the tRNA removal required for second-strand transfer reactions. PMID- 15333775 TI - A thermostable enzyme as an experimental platform to study properties of less stable homologues. AB - The structural and functional characterization of proteins is frequently hampered by lack of stability or by insufficient assembly of oligomeric proteins in over expression systems. Using F(1)-ATPase as a case study, we tackled this problem by introducing function-determining domains from a difficult-to-handle variety of an enzyme into a stable homologue. PMID- 15333776 TI - Focus on ART. PMID- 15333777 TI - Derivation, growth and applications of human embryonic stem cells. AB - Human embryonic stem (hES) cells are pluripotent cells derived from the inner cell mass cells of blastocysts with the potential to maintain an undifferentiated state indefinitely. Fully characterised hES cell lines express typical stem cell markers, possess high levels of telomerase activity, show normal karyotype and have the potential to differentiate into numerous cell types under in vitro and in vivo conditions. Therefore, hES cells are potentially valuable for the development of cell transplantation therapies for the treatment of various human diseases. However, there are a number of factors which may limit the medical application of hES cells: (a) continuous culture of hES cells in an undifferentiated state requires the presence of feeder layers and animal-based ingredients which incurs a risk of cross-transfer of pathogens; (b) hES cells demonstrate high genomic instability and non-predictable differentiation after long-term growth; and (c) differentiated hES cells express molecules which could cause immune rejection. In this review we summarise recent progress in the derivation and growth of undifferentiated hES cells and their differentiated progeny, and the problems associated with these techniques. We also examine the potential use of the therapeutic cloning technique to derive isogenic hES cells. PMID- 15333778 TI - Mitochondria in human oogenesis and preimplantation embryogenesis: engines of metabolism, ionic regulation and developmental competence. AB - Mitochondria are the most abundant organelles in the mammalian oocyte and early embryo. While their role in ATP production has long been known, only recently has their contribution to oocyte and embryo competence been investigated in the human. This review considers whether such factors as mitochondrial complement size, mitochondrial DNA copy numbers and defects, levels of respiration, and stage-specific spatial distribution, influence the developmental normality and viability of human oocytes and preimplantation-stage embryos. The finding that mitochondrial polarity can differ within and between oocytes and embryos and that these organelles may participate in the regulation of intracellular Ca(2+)homeostasis are discussed in the context of how focal domains of differential respiration and intracellular-free Ca(2+)regulation may arise in early development and what functional implications this may have for preimplantation embryogenesis and developmental competence after implantation. PMID- 15333779 TI - Deadly decisions: the role of genes regulating programmed cell death in human preimplantation embryo development. AB - Human preimplantation embryo development is prone to high rates of early embryo wastage, particularly under current in vitro culture conditions. There are many possible underlying causes for embryo demise, including DNA damage, poor embryo metabolism and the effect of suboptimal culture media, all of which could result in an imbalance in gene expression and the failed execution of basic embryonic decisions. In view of the complex interactions involved in embryo development, a thorough understanding of these parameters is essential to improving embryo quality. An increasing body of evidence indicates that cell fate (i.e. survival/differentiation or death) is determined by the outcome of specific intracellular interactions between pro- and anti-apoptotic proteins, many of which are expressed during oocyte and preimplantation embryo development. The recent availability of mutant mice lacking expression of various genes involved in the regulation of cell survival has enabled rapid progress towards identifying those molecules that are functionally important for normal oocyte and preimplantation embryo development. In this review we will discuss the current understanding of the regulation of cell death gene expression during preimplantation embryo development, with a focus on human embryology and a discussion of animal models where appropriate. PMID- 15333780 TI - Primate models for assisted reproductive technologies. AB - Although the deliberate creation of human embryos for scientific research is complicated by ethical and practical issues, a detailed understanding of the cellular and molecular events occurring during human fertilization is essential, particularly for understanding infertility. It is clear from cytoskeletal imaging studies of mouse fertilization that this information cannot be extrapolated to humans because of unique differences in centrosomal inheritance. However, the cytoskeletal rearrangements during non-human primate fertilization are very similar to humans, providing a compelling animal model in which to examine sperm- egg interactions. In order to address this key step in primate fertilization and to avoid the complexities in working with fertilized human zygotes, studies are now exploring the molecular foundations of various assisted fertilization techniques in a monkey model. While intracytoplasmic sperm injection with ejaculated or testicular sperm is quite successful in primate models, there are some specific differences when compared with standard IVF that warrant further investigation, particularly in regards to nuclear remodeling, genomic imprinting, Y-chromosome deletions and developmental outcomes. Similarly, primate models have been useful for examining spermatid function during fertilization but these have met with limited success. One area of primate reproductive research that has yet to be mastered is reproductive cloning. Genetically identical primates would provide the ultimate approach for accelerating stem cell-based therapies for a number of neurodegenerative diseases such as Alzheimer's and Parkinson's disease, as well as targeted gene therapies for various metabolic disorders. PMID- 15333781 TI - Effects of embryo culture on global pattern of gene expression in preimplantation mouse embryos. AB - Culture of preimplantation embryos affects gene expression. The magnitude of the effect on the global pattern of gene expression, however, is not known. We compared global patterns of gene expression in blastocysts cultured from the one cell stage in either Whitten's medium or KSOM + amino acids (KSOM/AA) with that of blastocysts that developed in vivo, using the Affymetrix MOE430A chip. The analysis revealed that expression of 114 genes was affected after culture in Whitten's medium, whereas only 29 genes were mis-expressed after culture in KSOM/AA. Expression Analysis Systematic Explorer was used to identify biological and molecular processes that are perturbed after culture and indicated that genes involved in protein synthesis, cell proliferation and transporter function were down-regulated after culture in Whitten's medium. A common set of genes involved in transporter function was also down-regulated after culture in KSOM/AA. These results provide insights as to why embryos develop better in KSOM/AA than in Whitten's medium, and highlight the power of microarray analysis to assess global patterns of gene expression. PMID- 15333782 TI - Cumulus expansion and glucose utilisation by bovine cumulus-oocyte complexes during in vitro maturation: the influence of glucosamine and follicle-stimulating hormone. AB - Glucose is an important metabolite and its presence during in vitro oocyte maturation (IVM) can have profound effects on the oocyte's developmental capacity. We have demonstrated that glucose uptake increases over a 24 h IVM period, with most accounted for as l-lactate production. However, as maturation proceeds, l-lactate production remains constant, suggesting an alternative role for glucose metabolism. We hypothesised that in the latter stages of oocyte maturation, glucose not accounted for by l-lactate production is utilised for FSH stimulated extracellular matrix (ECM) synthesis. To examine precursor utilisation for synthesis of ECM, bovine cumulus-oocyte complexes (COCs) were matured in +/- FSH and/or glucosamine (an alternative substrate of matrix components). Measurements included COC diameters, glucose consumption and l-lactate production in spent media and [U-(14)C]glucose incorporation into ECM. FSH significantly stimulated both diameter and glucose consumption during 20-24 h maturation compared with unstimulated complexes, although co-incubation with glucosamine and FSH decreased total glucose consumption 1.7-fold compared with FSH alone (P < 0.05). Furthermore, there was a linear relationship between glucose and l-lactate metabolism in the presence of glucosamine, suggesting that the majority of glucose was being utilised for l-lactate production via glycolysis. In the presence of glucosamine, twofold less [U-(14)C]glucose was incorporated into matrix compared with COCs cultured without glucosamine. These results support the hypothesis that there is a link between glucose and glucosamine uptake in FSH stimulated ECM synthesis. Furthermore, glucose has multiple fates within the COC during maturation and levels of utilisation are dependent on the composition of the maturation environment. PMID- 15333783 TI - Insulin-like growth factors-I and -II and insulin-like growth factor-binding protein-2 in dominant equine follicles during spring transition and the ovulatory season. AB - The period between seasonal anoestrus and cyclicity is characterized in many mares by cyclical growth and regression of large dominant follicles. The insulin like growth factor (IGF) system plays a key role in follicular growth and regression; therefore, we hypothesized that changes in the IGF system and its binding proteins would modulate onset of cyclicity in mares. Ovaries were obtained from pony mares on the day after detection of an actively growing 30 mm transitional anovulatory follicle, and also at the second or third oestrus of the breeding season on the day after the preovulatory follicle reached 30 mm in diameter. Size of dominant follicles at the time of removal was similar in transition (32 +/- 0.8 mm) and at oestrus (34 +/- 0.6 mm). IGF-I mRNA was present in granulosa cells, with low thecal expression, whereas IGF-II mRNA was confined to the theca layer. Expression of IGF-I and -II mRNAs, and intrafollicular concentrations of oestradiol, were lower (P < 0.01; paired t test) in transitional anovulatory follicles than in preovulatory follicles. Messenger RNA encoding IGFBP-2 was present in both theca and granulosa layers. Steady-state concentrations of mRNA encoding IGFBP-2 mRNA increased (P < 0.001) in theca in preovulatory follicles. Intrafollicular concentrations of IGFBP-2 were higher (P < 0.001) in transitional than in preovulatory follicles. The similarity in circulating concentrations of IGF-I in transitional and cyclic mares, suggested that the somatotrophic axis is not involved in transition from anovulatory to ovulatory cycles. The results suggest that the increased expression of IGF-I and II mRNAs in preovulatory follicles, along with the decrease in IGFBP-2 concentrations, could increase the bioavailability of intrafollicular IGF in large follicles during the breeding season, and support our hypothesis that intrafollicular IGF bioavailability must exceed a threshold level before ovulation can occur. PMID- 15333784 TI - Immunohistochemical localization of androgen receptors in the urogenital tracts of human embryos. AB - The aim of this study was to investigate androgen receptor (AR) expression in the developing human urogenital tract. The distribution of AR was examined in paraffin-embedded tissue sections of the lower urogenital tract using 55 human embryos of 8-12 weeks of gestation. Immunohistochemistry was performed for AR detection and gender was determined by polymerized chain reaction. There were no differences in the distribution of AR in male and female embryos at any stage of gestation. AR was present only in the mesenchymal tissues of the urogenital sinus at 8 weeks whilst the epithelium was negative, but after 9 weeks the epithelium also showed progressively more positive staining. In the phallus, AR staining was prominent. There was far less staining in the epithelium of the urethral groove from 8 to 10 weeks, whilst the mesenchyme of the urethral folds showed positive staining. At 11 and 12 weeks, both the urethral groove and folds showed uniform staining. The genital tubercle, genital swelling and bulbourethral gland precusors were also positively stained, although paramesonephric ducts were negative. Staining was observed in the mesonephric duct from 9 weeks. There was an absence of staining in the rectum at all stages of gestation. The expression of AR in an epithelium may be dependent upon the mesenchyme. Mesenchymal epithelial interactions played an important role in development, as has been described in experimental animals. AR expression could play a part in the growth of the genital organs. PMID- 15333785 TI - Evidence of increased substrate availability to in vitro-derived bovine foetuses and association with accelerated conceptus growth. AB - Changes in placental development have been associated with foetal abnormalities after in vitro embryo manipulations. This study was designed to investigate bovine conceptus development and substrate levels in plasma and fluids in in vivo and in vitro-produced (IVP) concepti and neonates. In vivo-produced and IVP embryos were derived by established embryo production procedures. Pregnant animals from both groups were slaughtered on days 90 or 180 of gestation, or allowed to go to term. Conceptus and neonatal physical traits were recorded; foetal, maternal and neonatal blood, and foetal fluids were collected for the determination of blood and fluid chemistry, and glucose, fructose and lactate concentrations. Placental transcripts for specific glucose transporters were determined by quantitative RT-PCR. No significant differences in uterine and conceptus traits were observed between groups on day 90. On day 180, larger uterine, placental and foetal weights, and an increase in placental gross surface area (SA) in IVP pregnancies were associated with increased glucose and fructose accumulation in foetal plasma and associated fluids, with no differences in the expression of components of the glucose transporter system. Therefore, the enlarged placental SA in IVP pregnancies suggests an increase in substrate uptake and transport capacity. Newborn IVP calves displayed higher birth weights and plasma fructose concentrations soon after birth, findings which appeared to be associated with clinical and metabolic distress. Our results indicated larger concepti and increased placental fructogenic capacity in mid- to late IVP pregnancies, features which appeared to be associated with an enhanced substrate supply, potentially glucose, to the conceptus. PMID- 15333786 TI - EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways. AB - Epidermal growth factor (EGF) is present in the maternal-fetal environment and has an important role in placental development. Matrix metalloproteinase-9 (MMP 9) expression/activation is a pre-requisite in extravillous trophoblast invasion. Whereas EGF up-regulates MMP-9 activity in a variety of cell types, there is no direct evidence for the stimulation of MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1) secretion by EGF in extravillous trophoblasts. In addition, the signalling pathways involved in this regulation are not clear. In the present study, we have examined the possible involvement of the phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways in the regulation of the MMP-9/TIMP-1 system by EGF in vitro. We used a well-established invasive extravillous trophoblast cell line (HTR8/Svneo) and measured gene and protein expression by semi-quantitative RT-PCR and western analysis respectively. MMP activity was determined by zymography. We showed for the first time that EGF activated both PI3K/Akt and MAPK/extracellular-signal regulated kinase (ERK) signalling in HTR8/SVneo, and increased both MMP-9 and TIMP-1 mRNAs and protein concentrations. Interfering with either signalling pathway via PI3K inhibitor LY294002 or MEK inhibitor U0126 in EGF-stimulated HTR8/SVneo cells blocked the induction of MMP-9 and TIMP-1. LY294002 inhibited Akt phosphorylation, but had no effect on ERK phosphorylation; U0126 suppressed ERK phosphorylation without interfering with the phosphorylation of Akt. In addition, expression of constitutively active Akt (Myr-Akt1, Myr-Akt2, Myr-Akt3) was not sufficient to induce proMMP-9 and TIMP-1 secretion. Our results suggest that the activation of both PI3K and MAPK pathways in extravillous trophoblasts is necessary for the up-regulation of MMP-9 and TIMP-1 expression by EGF. PMID- 15333787 TI - Changes in the reproductive system of male mice immunized with a GnRH-analogue conjugated to mycobacterial hsp70. AB - Immunosterilization is an attractive alternative to surgical castration. Gonadotropin-releasing hormone (GnRH) controls the production of the gonadotropins thereby having an orchestrating effect on the reproductive hormone cascade and spermatogenesis. Induction of neutralizing antibody can abrogate the effect of the hormone. Current GnRH-based vaccines often require strong adjuvants and/or multiple injections of the vaccines to overcome variability in the response. Heat shock proteins (hsp) have been used as carrier molecules because of their powerful intrinsic ability to enhance an immune response to associated antigens. A GnRH-analogue, GnRH-d6-Lys, was conjugated to recombinant Mycobacterium tuberculosis hsp70. Male BALB/c mice were immunized i.p. with GnRH hsp70 in the mild adjuvant Ribi or in incomplete Freund's adjuvant (IFA). The initial immunizations were done on pre-pubertal 3-week-old mice, with boosts at 5 and 8 weeks of age. The mice were killed at 10 weeks of age and GnRH-specific antibodies and serum testosterone levels measured. All the immunized mice mounted GnRH-specific antibody responses, with no difference in the mice immunized with GnRH-hsp70/Ribi or with GnRH-hsp70/IFA. There was substantial atrophy of the urogenital complex and significantly (P < 0.0005) reduced levels of testosterone dependent testicular relaxin-like factor mRNA expression. Mice immunized with GnRH-hsp70/Ribi showed substantially reduced (P < 0.001) serum testosterone levels. These results indicate that hsp70 may serve as a particularly advantageous carrier for GnRH-based vaccines. PMID- 15333788 TI - Effects of cyclic steroid hormone replacement on prolactin and luteinizing hormone surges in female rats. AB - The ability of steroid hormones to produce an LH or prolactin (PRL) surge was determined in rats ovariectomized at 6, 9 or 13 weeks of age and subjected to one, three or six cycles of estrogen and progesterone replacement. Sensitivity to steroid replacement was dependent on the age of the animal at the time of ovariectomy. Repeated cyclic steroid hormone replacement significantly increased the magnitude of the PRL response, but not the LH response, in animals ovariectomized at 6 weeks. The LH response was significantly altered by cyclic steroid replacement only in animals ovariectomized at 13 weeks. These results indicate that the mechanisms involved in the regulation of PRL secretion are influenced by steroid hormone replacement and that cyclic steroid hormone exposure increases the magnitude of the PRL secretory response. PMID- 15333789 TI - Can in vivo assessment of tissue hemodynamics with dynamic contrast-enhanced CT be used in the diagnosis of tumors and monitoring of cancer therapy outcomes? PMID- 15333790 TI - Vertebroplasty and kyphoplasty: do fluoroscopy operators know about radiation dose, and should they want to know? PMID- 15333791 TI - Assessment of fat content in supraspinatus muscle with proton MR spectroscopy in asymptomatic volunteers and patients with supraspinatus tendon lesions. AB - PURPOSE: To evaluate proton magnetic resonance (MR) spectroscopy in the assessment of lipid content of the supraspinatus muscle in asymptomatic volunteers and patients with supraspinatus tendon lesions. MATERIALS AND METHODS: Single-voxel proton MR spectroscopy was used to assess lipid content of the supraspinatus muscle in asymptomatic volunteers (n = 30) and patients with partial-thickness supraspinatus tendon tears (n = 30), newly diagnosed full thickness supraspinatus tendon tears (n = 30), and chronic full-thickness supraspinatus tendon tears (n = 30). The apparent lipid content of the supraspinatus muscle measured with proton MR spectroscopy was related to its appearance on sagittal-oblique T1-weighted spin-echo MR images (grades 0-4). One way analysis of variance was performed to test for significant differences, and the Tukey honestly significant difference procedure was performed for post hoc comparisons. RESULTS: Mean apparent lipid content was 13.7% (95% confidence interval [CI]: 11.5%, 15.8%) for asymptomatic volunteers, 29.5% (95% CI: 25.1%, 34.0%) for patients with partial-thickness tears, 48.6% (95% CI: 41.3%, 55.9%) for patients with full-thickness tears, and 66.1% (95% CI: 57.7%, 74.5%) for patients with chronic tears. Values were significantly different (analysis of variance, P <.001; P <.001-.002 for all post hoc pairwise comparisons). Mean apparent lipid content for the supraspinatus muscle was as follows: grade 0, 19.6% (95% CI: 16.7%, 22.6%); grade 1, 36.8% (95% CI: 33.2%, 40.4%); grade 2, 53.6% (95% CI: 43.1%, 64.2%); grade 3, 67.5% (95% CI: 52.6%, 82.3%); and grade 4, 79.2% (95% CI: 73.2%, 85.3%). With analysis of variance (P <.001), all post hoc pairwise comparisons were significant (P .05). Mean qualitative enhancement scores for interactive and standard protocol groups were 4.47 and 4.44, respectively (P =.47). CONCLUSION: The interactive method is a simple, efficient, and reproducible way to optimize aortoiliac enhancement while reducing contrast medium dose. PMID- 15333800 TI - Peripheral vascular disease: blinded study of dedicated calf MR angiography versus standard bolus-chase MR angiography and film hard-copy angiography. AB - PURPOSE: To compare the accuracy of contrast material-enhanced three-dimensional (3D) dedicated calf magnetic resonance (MR) angiography with that of bolus-chase MR angiography, with conventional angiography as the reference standard, in patients with symptomatic peripheral vascular disease (PVD). MATERIALS AND METHODS: Thirty men with symptomatic PVD were examined. MR angiography was performed at 1.5 T before conventional angiography. MR angiographic examination included 3D contrast-enhanced dedicated calf MR angiography and three-station bolus-chase MR angiography. Two radiologists blinded to conventional angiographic results evaluated the MR angiograms independently. Two angiographers evaluated the conventional angiograms in consensus. Calf artery segments were graded as having 50% or less stenosis, greater than 50% stenosis, or occlusion or as being nondiagnostic. Statistical analyses were performed with paired permutation testing. RESULTS: Analyses of 472 calf segments and 420 pelvic and thigh segments were performed. Of the 472 calf segments, three and 75 segments (reader 1) and seven and 91 segments (reader 2) were graded as nondiagnostic at dedicated calf MR angiography and bolus-chase MR angiography, respectively. Differences in diagnostic grade between the two examinations were significant (P <.001), accounting for within-subject correlations, with a mean estimated difference of 17.1% (95% confidence interval [CI]: -25.8%, -8.4%). In the calf arteries, the dedicated and bolus-chase MR angiographic sequences had diagnostic accuracies, respectively, of 81.5% (reader 1) and 79.1% (reader 2) and of 67.8% (reader 1) and 63.4% (reader 2). The dedicated calf sequence was significantly more accurate than the bolus-chase sequence (P =.001). The point estimate of the difference was 14.7%, with estimated correct diagnosis rates of 80.3% and 65.6% for the dedicated calf and bolus-chase examinations, respectively (95% CI for difference: 4.0%, 25.4%). The diagnostic accuracy of bolus-chase MR angiography at the pelvis thigh level was slightly higher when it was performed first: 81.9% (reader 1) and 83.8% (reader 2) versus 74.3% (reader 1) and 80.0% (reader 2) when it was performed last. The difference was not significant (P =.21). CONCLUSION: Use of dedicated calf MR angiography led to significantly increased diagnostic accuracy in the calf arteries compared with standard bolus-chase MR angiography. Use of the dual-bolus technique did not jeopardize the diagnostic accuracy in the pelvic and thigh arteries. PMID- 15333801 TI - Obstructive sleep apnea: MR imaging volume segmentation analysis. AB - PURPOSE: To retrospectively determine airway wall motion with volume segmentation of transverse cine magnetic resonance (MR) images in children with obstructive sleep apnea (OSA). MATERIALS AND METHODS: Transverse fast gradient-echo cine MR images of the hypopharynx were obtained at 1.5 T in 31 children with OSA (eight girls, 23 boys; mean age, 11.3 years) and 21 children free of airway symptoms who underwent MR imaging for other clinical indications (11 girls, 10 boys; mean age, 3.5 years). Volume segmentation with a k-means clustering algorithm was applied to transverse cine MR images to quantify airway volumes at each time. Airway wall motion for each child was described with standard deviation and range. Coefficient of variance and normalized range, which are independent of airway size, were used to compare groups (Kruskal-Wallis test). RESULTS: Plots of airway volume over time demonstrated large fluctuations during respiration in children with OSA and minimal fluctuations in controls; findings were consistent with airway distention and airway collapse in OSA. Average airway transverse volume was larger in the group with OSA than in the control group (OSA group, 2.52 mL; control group, 0.936 mL; P <.001). Mean standard deviation (OSA group, 0.840 mL; control group, 0.17 mL; P <.001) and mean range of airway cross section (OSA group, 3.552 mL; control group, 0.864 mL; P <.001) were larger in the group with OSA. Coefficient of variance (OSA group, 0.32; control group, 0.17; P <.001) and normalized range (OSA group, 1.42; control group, 0.96; P <.001) indicate statistically significant difference in airway dynamics in children with OSA. CONCLUSION: Volume segmentation of transverse cine MR images of the hypopharynx aids in quantification of increased airway wall motion in children with OSA. Transverse MR imaging demonstrates both airway distention and collapse in children with OSA. PMID- 15333802 TI - MR-guided vacuum-assisted breast biopsy: accuracy of targeting and success in sampling in a phantom model. AB - An 11-gauge MR-compatible system was designed for use in magnetic resonance vacuum-assisted breast biopsy. The system uses a detachable needle with minimal artifact to allow imaging after placement and before biopsy to confirm lesion location. A phantom study involving 16 biopsies of lesions smaller than 10 mm was conducted to assess the performance of this system. Fifteen (94%) of 16 biopsies resulted in successful lesion sampling. Eighty-five percent of all core samples contained specimen materials targeted lesion material. Further research to evaluate the efficiency, accuracy, and safety of this system in a patient population is recommended. PMID- 15333803 TI - Percutaneous radiofrequency ablation of pulmonary malignancies: giving credit for previous work. PMID- 15333805 TI - Systematic review of studies on elective endovascular repair versus conventional surgery for abdominal aortic aneurysms. PMID- 15333804 TI - Electric influence of NaCl concentration into the tissue in radiofrequency ablation. PMID- 15333806 TI - Oral use of gadopentetate dimeglumine for anastomotic leak in patients with iodine sensitivity. PMID- 15333808 TI - Academic health II. PMID- 15333809 TI - Pharmacogenomics. Cancer sharpshooters rely on DNA tests for a better aim. PMID- 15333810 TI - U.S. visa policy. Foreign scholars to get longer clearance. PMID- 15333811 TI - Next linear collider. Physicists pick a cold road for accelerator project. PMID- 15333812 TI - Chemistry. Fuel cell draws power from poison. PMID- 15333813 TI - Genetics. Patient advocate named co-inventor on patent for the PXE disease gene. PMID- 15333814 TI - Nuclear weapons policy. Showdown expected in Congress. PMID- 15333815 TI - Primate studies. Politics derail European chimp home. PMID- 15333816 TI - Biosecurity. Up in the air. PMID- 15333817 TI - Society for Conservation Biology meeting. Loss of dung beetles puts ecosystems in deep doo-doo. PMID- 15333818 TI - Society for Conservation Biology meeting. Forest loss makes monkeys sick. PMID- 15333819 TI - John Schaefer profile. Shooting for the stars. PMID- 15333820 TI - John Schaefer profile. The desire to go faint, fast. PMID- 15333821 TI - Ecology. Sportfishers on the hook for dwindling U.S. fish stocks. PMID- 15333822 TI - Finding evidence for black holes. PMID- 15333824 TI - Disagreements over cloud absorption. PMID- 15333823 TI - Extending life-span in C. elegans. PMID- 15333825 TI - Public health. Whatever happened to the U.S. AIDS epidemic? PMID- 15333826 TI - Neuroscience. In the place space. PMID- 15333827 TI - Behavior. Sweet revenge? PMID- 15333828 TI - Geoscience. What caused the great Lisbon earthquake? PMID- 15333829 TI - Geoscience. Tidal triggering caught in the act. PMID- 15333830 TI - Chiral-selective aminoacylation of an RNA minihelix. AB - Amino acids in natural proteins have a chiral, asymmetric center at the alpha carbon that is of the L-configuration. The sugar backbone of natural RNAs are also homochiral, but of the D-configuration. Because protein synthesis requires the aminoacylation of RNA, it is this step that could have provided chiral selectivity. Here we show that an RNA minihelix was aminoacylated by an aminoacyl phosphate-D-oligonucleotide with a clear preference for L- as opposed to D-amino acids. A mirror-image RNA system showed the opposite selectivity. These results suggest the possibility that the selection of L-amino acids for proteins was determined by the stereochemistry of RNA. PMID- 15333832 TI - Spatial representation in the entorhinal cortex. AB - As the interface between hippocampus and neocortex, the entorhinal cortex is likely to play a pivotal role in memory. To determine how information is represented in this area, we measured spatial modulation of neural activity in layers of medial entorhinal cortex projecting to the hippocampus. Close to the postrhinal-entorhinal border, entorhinal neurons had stable and discrete multipeaked place fields, predicting the rat's location as accurately as place cells in the hippocampus. Precise positional modulation was not observed more ventromedially in the entorhinal cortex or upstream in the postrhinal cortex, suggesting that sensory input is transformed into durable allocentric spatial representations internally in the dorsocaudal medial entorhinal cortex. PMID- 15333831 TI - The neural basis of altruistic punishment. AB - Many people voluntarily incur costs to punish violations of social norms. Evolutionary models and empirical evidence indicate that such altruistic punishment has been a decisive force in the evolution of human cooperation. We used H2 15O positron emission tomography to examine the neural basis for altruistic punishment of defectors in an economic exchange. Subjects could punish defection either symbolically or effectively. Symbolic punishment did not reduce the defector's economic payoff, whereas effective punishment did reduce the payoff. We scanned the subjects' brains while they learned about the defector's abuse of trust and determined the punishment. Effective punishment, as compared with symbolic punishment, activated the dorsal striatum, which has been implicated in the processing of rewards that accrue as a result of goal-directed actions. Moreover, subjects with stronger activations in the dorsal striatum were willing to incur greater costs in order to punish. Our findings support the hypothesis that people derive satisfaction from punishing norm violations and that the activation in the dorsal striatum reflects the anticipated satisfaction from punishing defectors. PMID- 15333834 TI - Direct measurement of light waves. AB - The electromagnetic field of visible light performs approximately 10(15) oscillations per second. Although many instruments are sensitive to the amplitude and frequency (or wavelength) of these oscillations, they cannot access the light field itself. We directly observed how the field built up and disappeared in a short, few-cycle pulse of visible laser light by probing the variation of the field strength with a 250-attosecond electron burst. Our apparatus allows complete characterization of few-cycle waves of visible, ultraviolet, and/or infrared light, thereby providing the possibility for controlled and reproducible synthesis of ultrabroadband light waveforms. PMID- 15333833 TI - Search for low-mass exoplanets by gravitational microlensing at high magnification. AB - Observations of the gravitational microlensing event MOA 2003-BLG-32/OGLE 2003 BLG-219 are presented, for which the peak magnification was over 500, the highest yet reported. Continuous observations around the peak enabled a sensitive search for planets orbiting the lens star. No planets were detected. Planets 1.3 times heavier than Earth were excluded from more than 50% of the projected annular region from approximately 2.3 to 3.6 astronomical units surrounding the lens star, Uranus-mass planets were excluded from 0.9 to 8.7 astronomical units, and planets 1.3 times heavier than Saturn were excluded from 0.2 to 60 astronomical units. These are the largest regions of sensitivity yet achieved in searches for extrasolar planets orbiting any star. PMID- 15333835 TI - Nanoribbon waveguides for subwavelength photonics integration. AB - Although the electrical integration of chemically synthesized nanowires has been achieved with lithography, optical integration, which promises high speeds and greater device versatility, remains unexplored. We describe the properties and functions of individual crystalline oxide nanoribbons that act as subwavelength optical waveguides and assess their applicability as nanoscale photonic elements. The length, flexibility, and strength of these structures enable their manipulation on surfaces, including the optical linking of nanoribbon waveguides and other nanowire elements to form networks and device components. We demonstrate the assembly of ribbon waveguides with nanowire light sources and detectors as a first step toward building nanowire photonic circuitry. PMID- 15333836 TI - Transparent, conductive carbon nanotube films. AB - We describe a simple process for the fabrication of ultrathin, transparent, optically homogeneous, electrically conducting films of pure single-walled carbon nanotubes and the transfer of those films to various substrates. For equivalent sheet resistance, the films exhibit optical transmittance comparable to that of commercial indium tin oxide in the visible spectrum, but far superior transmittance in the technologically relevant 2- to 5-micrometer infrared spectral band. These characteristics indicate broad applicability of the films for electrical coupling in photonic devices. In an example application, the films are used to construct an electric field-activated optical modulator, which constitutes an optical analog to the nanotube-based field effect transistor. PMID- 15333837 TI - Powering fuel cells with CO via aqueous polyoxometalates and gold catalysts. AB - Electricity was produced by catalytic oxidation of carbon monoxide (CO) by using gold catalysts at room temperature. The observed rates are faster than conventional processes operating at 500 kelvin or higher for the conversion of CO with water to produce hydrogen and carbon dioxide through the water-gas shift (WGS). By eliminating the WGS reaction, we remove the need to transport and vaporize liquid water in the production of energy for portable applications. This process can use CO-containing gas streams from the catalytic reforming of hydrocarbons to produce an aqueous solution of reduced polyoxometalate compounds that can be used to generate power. The reduced polyoxometalate can be reoxidized in fuel cells that contain simple carbon anodes. PMID- 15333838 TI - Plasminogen is a critical host pathogenicity factor for group A streptococcal infection. AB - Group A streptococci, a common human pathogen, secrete streptokinase, which activates the host's blood clot-dissolving protein, plasminogen. Streptokinase is highly specific for human plasminogen, exhibiting little or no activity against other mammalian species, including mouse. Here, a transgene expressing human plasminogen markedly increased mortality in mice infected with streptococci, and this susceptibility was dependent on bacterial streptokinase expression. Thus, streptokinase is a key pathogenicity factor and the primary determinant of host species specificity for group A streptococcal infection. In addition, local fibrin clot formation may be implicated in host defense against microbial pathogens. PMID- 15333839 TI - E protein silencing by the leukemogenic AML1-ETO fusion protein. AB - The AML1-ETO fusion protein, generated by the t(8;21) chromosomal translocation, is causally involved in nearly 15% of acute myeloid leukemia (AML) cases. This study shows that AML1-ETO, as well as ETO, inhibits transcriptional activation by E proteins through stable interactions that preclude recruitment of p300/CREB binding protein (CBP) coactivators. These interactions are mediated by a conserved ETO TAF4 homology domain and a 17-amino acid p300/CBP and ETO target motif within AD1 activation domains of E proteins. In t(8;21) leukemic cells, very stable interactions between AML1-ETO and E proteins underlie a t(8;21) translocation-specific silencing of E protein function through an aberrant cofactor exchange mechanism. These studies identify E proteins as AML1-ETO targets whose dysregulation may be important for t(8;21) leukemogenesis, as well as an E protein silencing mechanism that is distinct from that associated with differentiation-inhibitory proteins. PMID- 15333840 TI - Impaired degradation of mutant alpha-synuclein by chaperone-mediated autophagy. AB - Aberrant alpha-synuclein degradation is implicated in Parkinson's disease pathogenesis because the protein accumulates in the Lewy inclusion bodies associated with the disease. Little is known, however, about the pathways by which wild-type alpha-synuclein is normally degraded. We found that wild-type alpha-synuclein was selectively translocated into lysosomes for degradation by the chaperone-mediated autophagy pathway. The pathogenic A53T and A30P alpha synuclein mutants bound to the receptor for this pathway on the lysosomal membrane, but appeared to act as uptake blockers, inhibiting both their own degradation and that of other substrates. These findings may underlie the toxic gain-of-function by the mutants. PMID- 15333842 TI - The great indoors. PMID- 15333843 TI - TRPV1 and cough. PMID- 15333844 TI - Pneumocystis jirovecii infection. PMID- 15333845 TI - Location, location, location: microlocalisation of inflammatory cells and airway dysfunction. PMID- 15333846 TI - Journal impact factors for 2003: Thorax increases. PMID- 15333847 TI - Moran Campbell and clinical science. PMID- 15333848 TI - Effects of gas and other fume emitting heaters on the development of asthma during childhood. AB - BACKGROUND: Several studies have shown adverse effects of gas cookers and heaters on respiratory health. The long term effects of early life exposure to these appliances are not known. This study investigated the effect of exposure to fume emitting heaters, currently and during the first year of life, on the risk of asthma outcomes. METHODS: A cross sectional study of schoolchildren (n = 627) aged 8-11 years was conducted in Belmont, Australia. Information on symptoms and heating types was collected by parent completed questionnaire. Atopy was assessed by skin prick tests and airway hyperresponsiveness (AHR) was assessed by histamine challenge test. RESULTS: There was no association between the current use of fume emitting heaters and any of the asthma outcomes. However, having been exposed to fume emitting heaters during the first year of life was associated with an increased risk of having AHR (relative risk (RR) 1.47, 95% confidence interval (CI) 1.06 to 2.03), recent wheeze (RR 1.44, 95% CI 1.11 to 1.86), and recent wheeze + AHR (RR 2.08, 95% CI 1.31 to 3.31). CONCLUSION: If confirmed in other settings, this finding would require a review of the range of heating types that are appropriate for use in households in which young children live. PMID- 15333849 TI - Association of domestic exposure to volatile organic compounds with asthma in young children. AB - AIM: To investigate the association between domestic exposure to volatile organic compounds (VOCs) and asthma in young children. METHODS: A population based case control study was conducted in Perth, Western Australia in children aged between 6 months and 3 years. Cases (n = 88) were children recruited at Princess Margaret Hospital accident and emergency department and discharged with asthma as the primary diagnosis; 104 controls consisted of children from the same age group without an asthma diagnosis identified through the Health Department of Western Australia. Information regarding the health status of the study children and characteristics of the home was collected using a standardised questionnaire. Exposure to VOCs, average temperature and relative humidity were measured in winter and summer in the living room of each participating household. RESULTS: Cases were exposed to significantly higher VOC levels (microg/m3) than controls (p<0.01). Most of the individual VOCs appeared to be significant risk factors for asthma with the highest odds ratios for benzene followed by ethylbenzene and toluene. For every 10 unit increase in the concentration of toluene and benzene (microg/m3) the risk of having asthma increased by almost two and three times, respectively. CONCLUSIONS: Domestic exposure to VOCs at levels below currently accepted recommendations may increase the risk of childhood asthma. Measurement of total VOCs may underestimate the risks associated with individual compounds. PMID- 15333850 TI - Longitudinal study of grass pollen exposure, symptoms, and exhaled nitric oxide in childhood seasonal allergic asthma. AB - BACKGROUND: Exhaled nitric oxide (NO) has been proposed as a marker of airway eosinophilic inflammation in asthma. There is currently a paucity of longitudinal data relating it to allergen exposure and asthma symptoms. METHODS: Forty four children (6-16 years) with seasonal allergic asthma were sequentially followed before and during the grass pollen season. Asthma symptoms, lung function, NO levels, and pollen counts were recorded. The relationship between exhaled NO and both the pollen levels and asthma control were assessed longitudinally, comparing a subject's measurements with their previous ones. RESULTS: The median exhaled NO concentration was significantly increased during the pollen season (6.2 v 9.2 parts per billion (ppb), p<0.002; median change 2.9 ppb, 95% confidence interval 1.5 to 5.4). Exhaled NO was best associated with the mean pollen count in the week before measurement. It was also significantly associated with asthma control. CONCLUSIONS: The results suggest that, within a longitudinal model, the exhaled NO concentration is related to preceding allergen exposure and asthma control. It may be clinically more useful to compare exhaled NO values with a subject's previous values than to compare them with a population based normal range. PMID- 15333851 TI - Epithelial inducible nitric oxide synthase activity is the major determinant of nitric oxide concentration in exhaled breath. AB - BACKGROUND: The fractional concentration of nitric oxide (NO) in exhaled breath (FeNO) is increased in asthma. There is a general assumption that NO synthase (NOS) 2 in epithelium is the main source of NO in exhaled breath. However, there is no direct evidence to support the assumption and data from animal models suggest that non-inducible NOS systems have important roles in determining airway reactivity, regulating inflammation, and might contribute significantly to NO measured in exhaled breath. METHODS: Bronchial epithelial cells were obtained from healthy, atopic, and asthmatic children by non-bronchoscopic brushing. Exhaled NO (FeNO) was measured directly using a fast response chemiluminescence NO analyser. RNA was extracted from the epithelial cells and real time polymerase chain reaction was used to determine the expression of NOS isoenzymes. NOS2 was examined in macrophages and epithelial cells by immunohistochemistry. RESULTS: NOS1 mRNA was not detectable. NOS3 mRNA was detected in 36 of 43 samples at lower levels than NOS2 mRNA which was detectable in all samples. The median FeNO was 15.5 ppb (95% CI 10 to 18.1). There was a significant correlation between FeNO and NOS2 expression (R = 0.672, p<0.001). All epithelial cells exhibited NOS2 staining, whereas staining in the macrophages was variable and not related to phenotype. CONCLUSIONS: Only NOS2 expression was associated with FeNO in respiratory epithelial cells obtained from children (R = 0.672; p<0.001). This suggests that FeNO variability is largely determined by epithelial NOS2 expression with little contribution from other isoforms. PMID- 15333852 TI - Five year study of etidronate and/or calcium as prevention and treatment for osteoporosis and fractures in patients with asthma receiving long term oral and/or inhaled glucocorticoids. AB - BACKGROUND: Glucocorticoids are associated with a reduction in bone density and an increased risk of fracture. Concurrent treatment with bisphosphonates reduces bone loss and may prevent fractures. A randomised study was performed to determine whether treatment with cyclical etidronate and/or calcium for 5 years prevents fractures or reverses/reduces bone loss in patients receiving glucocorticoid treatment for asthma. METHODS: A multicentre, randomised, parallel group comparison of etidronate alone, calcium alone, etidronate + calcium, and no treatment, with stratification according to level of glucocorticoid exposure was carried out in 39 chest clinics in the UK. Three hundred and forty nine postmenopausal female and male outpatients with asthma aged 50-70 years were randomised. The main outcome measures were fractures and changes in bone mineral density (BMD). RESULTS: Overall, 8% of the patients experienced symptomatic fractures and 17.5% developed either a symptomatic fracture and/or a semiquantitative vertebral fracture by the end of 5 years There were no significant differences between the four treatment groups. Comparing etidronate with no etidronate, the rates of new fractures were not significantly different for symptomatic fractures (OR 1.07 (95% CI 0.46 to 2.47)) or for any fractures (OR 0.82 (95% CI 0.45 to 1.47)). For the comparison of calcium with no calcium the corresponding ORs were 1.43 (95% CI 0.62 to 3.33) and 0.91 (95% CI 0.50 to 1.63). In post hoc analysis the effect of etidronate was greater in women than in men (interaction p value 0.02) with the fracture incidence roughly halved (OR 0.39, 95% CI 0.14 to 0.99). Etidronate increased BMD at the lumbar spine by 4.1% (p = 0.001) while calcium had no significant effect. At the proximal femur the effects of treatment were not significant (relative increases etidronate 1.6%; calcium 1.1%). The rate of new fractures in patients with fractures at entry (23.7%) was higher than in those without fractures at entry (14.3%): OR 1.87 (95% CI 1.06 to 3.07). No association was found between change in BMD and new fractures. CONCLUSIONS: In patients receiving glucocorticoids for asthma etidronate significantly increased BMD over 5 years at the lumbar spine but not at the hip and had little if any protective effect against fractures, except possibly in postmenopausal women. The effects of calcium were not significant. Combination treatment had no advantage but increased unwanted effects. PMID- 15333853 TI - Antitussive activity of iodo-resiniferatoxin in guinea pigs. AB - BACKGROUND: Iodo-resiniferatoxin (I-RTX) has recently been described as an ultra potent antagonist of the transient receptor potential vanilloid-1 (TRPV1). METHODS: The ability of I-RTX to inhibit cough induced by inhalation of two putative TRPV1 stimulants (capsaicin and citric acid) was tested in non anaesthetised guinea pigs. RESULTS: Pretreatment with I-RTX either intraperitoneally (0.03-0.3 micromol/kg) or by aerosol (0.1-3 microM) reduced the number of coughs produced by inhalation of citric acid (0.25 M) and capsaicin (30 microM) in a dose dependent manner. Capsazepine (CPZ) also reduced citric acid and capsaicin induced cough, but the activity of I-RTX was 10-100 times more potent than CPZ in all the experimental conditions tested. CONCLUSIONS: I-RTX is a novel and potent antitussive drug which inhibits cough mediated by agents possibly acting via TRPV1 activation. PMID- 15333854 TI - Cough suppression during flexible bronchoscopy using combined sedation with midazolam and hydrocodone: a randomised, double blind, placebo controlled trial. AB - BACKGROUND: Current British Thoracic Society guidelines do not recommend routinely the combined use of a benzodiazepine and opiate during flexible bronchoscopy (FB). A randomised, placebo controlled, double blind study was undertaken to determine whether hydrocodone in combination with midazolan improves cough suppression during FB without increasing the risk of desaturation. METHODS: 120 patients were randomised to receive midazolam and 5 mg i.v. hydrocodone or midazolam and placebo with topical anaesthesia. Pulse oximetry was recorded continuously during FB. Bronchoscopists and nurses charted their perception of cough and the patients rated their discomfort during the procedure on a 10 cm visual analogue scale (VAS). RESULTS: There was no significant difference between the two groups with regard to the indication for FB, duration of procedure (21 (11) min v 22 (10) min, p = 0.570), doses of supplemental lignocaine (171 (60) mg v 173 (66) mg, p = 0.766) and midazolam (4.5 (2.3) mg v 4.9 (2.7) mg, p = 0.309), lowest oxygen saturation (94.8 (2.7) v 94.9 (2.7), p = 0.433), and desaturations < or =90%. Perception of cough by both the bronchoscopist and the nurse was significantly lower in the hydrocodone group (3 (0-10) and 3 (0-10)) than in the placebo group (6 (0-10) and 6 (0-10)), respectively (p = 0.001). According to the VAS scale, patients' tolerance was also significantly better with hydrocodone than with placebo (2 (0-8) v 3 (0-9), p = 0.043). CONCLUSION: The combination of midazolam and hydrocodone markedly reduces cough during FB without causing significant desaturation, especially when invasive diagnostic procedures are performed. PMID- 15333855 TI - Circulating cardiovascular risk factors in obstructive sleep apnoea: data from randomised controlled trials. AB - BACKGROUND: Obstructive sleep apnoea (OSA) is associated with high cardiovascular morbidity and mortality and is an independent risk factor for hypertension. Novel circulating cardiovascular risk markers enabling a more accurate prediction of cardiovascular risk have been identified. Examination of these markers may clarify the increased risk in OSA and contribute to an analysis of the benefits of treatment. METHODS: Plasma levels of total cholesterol and triglyceride and activated coagulation factors XIIa and VIIa, factors VII, VIII, XII, fibrinogen, thrombin-antithrombin (TAT), von Willebrand factor antigen (vWFAg), soluble P selectin (sP-sel), and homocysteine were measured before and after treatment for 1 month with therapeutic or subtherapeutic (control) continuous positive airways pressure (CPAP) in 220 patients with OSA. RESULTS: Levels of activated coagulation factors XIIa, VIIa, TAT and sP-sel were higher in OSA patients at baseline than in unmatched controls, but did not fall with 1 month of therapeutic CPAP treatment. The raised sP-sel levels correlated only with body mass index (p = 0.002). There was a trend towards a significant fall in total cholesterol with therapeutic CPAP (p = 0.06) compared with the control group. In the therapeutic group there was a clinically significant mean fall in total cholesterol of 0.28 mmol/l (95% confidence interval 0.11 to 0.45, p = 0.001) which may reduce cardiovascular risk by about 15%. CONCLUSION: A number of activated coagulation factors are increased in untreated OSA patients, potentially contributing to vascular risk, but they do not fall with 1 month of CPAP treatment. Nasal CPAP may produce a clinically relevant fall in total cholesterol level, potentially reducing cardiovascular risk, but this needs to be verified in a larger prospective study. PMID- 15333856 TI - Preferential reduction of quadriceps over respiratory muscle strength and bulk after lung transplantation for cystic fibrosis. AB - BACKGROUND: In the absence of complications, recipients of lung transplants for cystic fibrosis have normal pulmonary function but the impact of the procedure on the strength and bulk of respiratory and limb muscles has not been studied. METHODS: Twelve stable patients who had undergone lung transplantation for cystic fibrosis 48 months earlier (range 8-95) and 12 normal subjects matched for age, height, and sex were studied. The following parameters were measured: standard lung function, peak oxygen uptake by cycle ergometry, diaphragm surface area by computed tomographic (CT) scanning, diaphragm and abdominal muscle thickness by ultrasonography, twitch transdiaphragmatic and gastric pressures, quadriceps isokinetic strength, and quadriceps cross section by CT scanning, and lean body mass. Diaphragm mass was computed from diaphragm surface area and thickness. RESULTS: Twitch transdiaphragmatic and gastric pressures, diaphragm mass, and abdominal muscle thickness were similar in the two groups but quadriceps strength and cross section were decreased by nearly 30% in the patients. Patients had preserved quadriceps strength per unit cross section but reduced quadriceps cross section per unit lean body mass. The cumulative dose of corticosteroids was an independent predictor of quadriceps atrophy. Peak oxygen uptake showed positive correlations with quadriceps strength and cross section in the two groups, but peak oxygen uptake per unit quadriceps strength or cross section was reduced in the patient group. CONCLUSIONS: The diaphragm and abdominal muscles have preserved strength and bulk in patients transplanted for cystic fibrosis but the quadriceps is weak due to muscle atrophy. This atrophy is caused in part by corticosteroid therapy and correlates with the reduction in exercise capacity. PMID- 15333857 TI - Sequential photodynamic therapy (PDT) and high dose brachytherapy for endobronchial tumour control in patients with limited bronchogenic carcinoma. AB - BACKGROUND: Bulky endobronchial tumours in patients with lung cancer are difficult to treat. Brachytherapy and photodynamic therapy (PDT) are variably effective, and the combination of these treatments is not often recommended. However, cell culture studies and animal studies indicate a possible synergistic effect of combining PDT with ionising radiation. We assessed the safety and effectiveness of combined brachytherapy and PDT in patients with bulky endobronchial lung cancer. METHODS: Patients with histologically proven non-small cell bronchogenic carcinoma and bulky endobronchial tumours were treated using a combination of PDT (Photofrin, 2 mg/kg) and brachytherapy. Six weeks after PDT, brachytherapy was applied with five fractions of 4 Gy at weekly intervals. Follow up was performed with standard and autofluorescence bronchoscopy and tissue biopsies every 3 months. RESULTS: Thirty two patients were treated. Tumours were extensive with lengths ranging from 10 to 60 mm along the bronchus and estimated volumes ranging from 40 to 3500 mm3. At a mean follow up of 24 months, 26 patients were free of residual tumour and local recurrence. The remaining patients received a second treatment with PDT, brachytherapy, Nd:YAG laser coagulation, or external beam radiation. Distant metastases (lung, lymph node) developed in two of the six patients. Currently, all 32 patients are well. There is no evidence of residual or local recurrent endobronchial cancer in 28 patients and none had severe complications. CONCLUSION: The combination of PDT and brachytherapy for treating patients with lung cancer and extensive endobronchial tumour is safe and, in this study, had excellent therapeutic efficacy. PMID- 15333858 TI - Is endosonography guided fine needle aspiration (EUS-FNA) for sarcoidosis as good as we think? AB - BACKGROUND: Preliminary data show that endosonography guided fine needle aspiration (EUS-FNA) may be an accurate method for diagnosing sarcoidosis. However, these data were obtained in a small selected group of patients with a very high pretest probability of sarcoidosis. This retrospective study reports on the use of EUS-FNA in an unselected group of patients with mediastinal lymphadenopathy of unknown origin. METHODS: The EUS database of a single tertiary referral centre was reviewed for patients who underwent EUS-FNA for mediastinal lymphadenopathy of unknown origin. Clinical presentation and imaging studies of each case were carefully reviewed and the diagnosis "sarcoidosis" or "no sarcoidosis" attributed if possible. The diagnoses were compared with the result of EUS-FNA. RESULTS: One hundred and twenty four patients were investigated. In 35 cases EUS-FNA identified granulomas (group 1); in the other 89 cases (group 2) no granulomas were detected. The definite diagnoses in group 1 were sarcoidosis (n = 25), indefinite (n = 7), no sarcoidosis (n = 3). The definite diagnoses in group 2 were sarcoidosis (n = 3), indefinite (n = 9), no sarcoidosis (n = 77). Of the 77 cases with no sarcoidosis, 44 were diagnosed with other diseases. The other 33 showed non-specific changes in the FNA and sarcoidosis was excluded by negative non-EUS pathology (n = 17) and clinical presentation. The sensitivity and specificity for EUS-FNA were 89% (95% CI 82 to 94) and 96% (95% CI 91 to 98), respectively, after exclusion of the indefinite cases in both groups. CONCLUSIONS: EUS-FNA is an accurate method for diagnosing sarcoidosis in an unselected group of patients with mediastinal lymphadenopathy. The reported sensitivity and specificity must be appreciated in the context of the difficult and often incomplete clinical diagnosis of sarcoidosis. PMID- 15333859 TI - Survival and disease progression in UK patients with lymphangioleiomyomatosis. AB - BACKGROUND: Lymphangioleiomyomatosis (LAM) is a rare and progressive disease of young women with no effective treatment. Previous estimates of 10 year survival, based mostly on case series or patients from tertiary centres, have ranged from 40% to 79%; no data are available on the progression of respiratory disability. In order to provide data for patients and for planning intervention studies, we have looked at the time course of LAM using a national cohort. METHODS: Time to death, time to MRC dyspnoea grades 2-5, and need for oxygen in patients on the UK LAM database were analysed using Kaplan-Meier analysis and Cox regression. RESULTS: Fifty seven of 72 patients responded with a median duration of follow up of 12.6 years (range 2.3-37) from the onset of symptoms. Ten year survival was 91% from onset of symptoms but varied widely with 11 patients alive after 20 years. Median time to MRC grade 3 dyspnoea (breathless walking on the flat) was 9.3 years (95% CI 5.1 to 13.4) from onset of symptoms. CONCLUSIONS: Survival from LAM appears to be better than that reported in early studies. These data should be helpful for patients and for planning clinical trials. PMID- 15333860 TI - Sleep. 5: Driving and automobile crashes in patients with obstructive sleep apnoea/hypopnoea syndrome. AB - Driving is a complex task involving distinct cognitive, perceptual, motor, and decision making skills. After placing the vehicle on the road, the driver must constantly survey the ever changing roadway environment to keep the vehicle in the lane and moving at an appropriate safe speed. This surveillance involves two distinct visual tasks: estimating and responding to the oncoming curvature and controlling lane position. Driving is therefore a divided attention task involving speed and lane control as well as monitoring. To do this in a safe manner requires careful attention and alertness which can be problematic for patients with obstructive sleep apnoea/hypopnoea syndrome (OSAHS) or other sleep disorders. PMID- 15333862 TI - A case of lymphomatoid granulomatosis masquerading as a lung abscess. AB - Lymphomatoid granulomatosis (LG) is a rare T cell rich, B cell non-Hodgkin's lymphoma which is difficult to diagnose. We present a patient with LG who demonstrated many of the difficulties in diagnosis and highlighted the importance of reviewing the diagnosis if treatment does not have the anticipated effect. PMID- 15333863 TI - Lymphangioma: a rare cause of a mediastinal mass. PMID- 15333864 TI - POP goes the power wall? Taking aim at tobacco promotional strategies utilised at retail. PMID- 15333861 TI - Respiratory muscle fibres: specialisation and plasticity. AB - Skeletal muscles are composed of fibres of different types, each type being identified by the isoform of myosin heavy chain which is expressed as slow 1, fast 2A, fast 2X, and fast 2B. Slow fibres are resistant to fatigue due to their highly oxidative metabolism whereas 2X and 2B fibres are easily fatiguable and fast 2A fibres exhibit intermediate fatigue resistance. Slow fibres and fast fibres are present in equal proportions in the adult human diaphragm while intercostal muscles contain a higher proportion of fast fibres. A small fibre size, abundance of capillaries, and a high aerobic oxidative enzyme activity are typical features of diaphragm fibres and give them the resistance to fatigue required by their continuous activity. Because of their fibre composition, intercostal muscles are less resistant to fatigue. The structural and functional characteristics of respiratory muscle fibres are not fixed, however, and can be modified in response to several physiological and pathological conditions such as training (adaptation to changes in respiratory load), adaptation to hypoxia, age related changes, and changes associated with respiratory diseases. The properties of respiratory muscle fibres can also be modified by pharmacological agents such as beta2 agonists and corticosteroids used for the treatment of respiratory diseases. PMID- 15333866 TI - Bangladesh: bikers against tobacco. PMID- 15333865 TI - Sweet and spicy flavours: new brands for minorities and youth. AB - The tobacco industry is now adding sweet and spicy flavours to its products in order to increase sales to youth and minorities PMID- 15333867 TI - Canada: chicanery in the chicanes. PMID- 15333869 TI - Vietnam: health wins at South-East Asia games. PMID- 15333870 TI - Sri lanka: BAT uses religion, ousts quit charity scheme. PMID- 15333871 TI - South Africa: courting success. PMID- 15333872 TI - FCTC: how will they keep pushing? PMID- 15333873 TI - Turkey: F1 keeps on coming. PMID- 15333874 TI - USA: nightingales sing at PM's AGM. PMID- 15333875 TI - Particulate matter from tobacco versus diesel car exhaust: an educational perspective. AB - BACKGROUND: Air pollution is a common alibi used by adolescents taking up smoking and by smokers uncertain about quitting. However, environmental tobacco smoke (ETS) causes fine particulate matter (PM) indoor pollution exceeding outdoor limits, while new engines and fuels have reduced particulate emissions by cars. Data comparing PM emission from ETS and a recently released diesel car are presented. METHODS: A 60 m3 garage was chosen to assess PM emission from three smouldering cigarettes (lit sequentially for 30 minutes) and from a TDCi 2000cc, idling for 30 minutes. RESULTS: Particulate was measured with a portable analyser with readings every two minutes. Background PM10, PM2.5, and PM1 levels (mean (SD)) were 15 (1), 13 (0.7), and 7 (0.6) microg/m3 in the car experiment and 36 (2), 28 (1), and 14 (0.8) microg/m3 in the ETS experiment, respectively. Mean (SD) PM recorded in the first hour after starting the engine were 44 (9), 31 (5), and 13 (1) microg/m3, while mean PM in the first hour after lighting cigarettes were 343 (192), 319 (178), and 168 (92) microg/m3 for PM(10), PM2.5, and PM1, respectively (p < 0.001, background corrected). CONCLUSIONS: ETS is a major source of PM pollution, contributing to indoor PM concentrations up to 10-fold those emitted from an idling ecodiesel engine. Besides its educational usefulness, this knowledge should also be considered from an ecological perspective. PMID- 15333876 TI - The tobacco industry's use of Wall Street analysts in shaping policy. AB - OBJECTIVE: To document how the tobacco industry has used Wall Street analysts to further its public policy objectives. METHODS: Searching tobacco documents available on the internet, newspaper articles, and transcripts of public hearings. RESULTS: The tobacco industry used nominally independent Wall Street analysts as third parties to support the tobacco industry's legislative agenda at both national and state levels in the USA. The tobacco industry has, for example, edited the testimony of at least one analyst before he testified to the US Senate Judiciary Committee, while representing himself as independent of the industry. CONCLUSION: The tobacco industry has used undisclosed collaboration with Wall Street analysts, as they have used undisclosed relationships with research scientists and academics, to advance the interests of the tobacco industry in public policy. PMID- 15333877 TI - Public health foundations and the tobacco industry: lessons from Minnesota. AB - OBJECTIVE: To investigate whether private foundations can be created in a way that will insulate them from attacks by the tobacco industry, using the Minnesota Partnership for Action Against Tobacco (MPAAT) as a case study. DESIGN: Information was collected from internal tobacco industry documents, court documents, newspapers, and interviews with health advocates and elected officials. RESULTS: The creation of MPAAT as an independent foundation did not insulate it from attacks by tobacco industry allies. During 2001-2002, MPAAT was repeatedly attacked by Attorney General Mike Hatch and major media, using standard tobacco industry rhetoric. This strategy of attack and demands for information were reminiscent of previous attacks on Minnesota's Plan for Nonsmoking and Health and the American Stop Smoking Intervention Study (ASSIST). MPAAT was ultimately forced to restructure its programme to abandon effective community norm change interventions around smoke-free policies and replace them with less effective individual cessation interventions. Neither MPAAT nor other health advocates mounted an effective public response to these attacks, instead relying on the insider strategy of responding in court. CONCLUSION: It is not possible to avoid attacks by the tobacco industry or its political allies. Like programmes administered by government agencies, tobacco control foundations must be prepared for these attacks, including a proactive plan to educate the public about the principles of community based tobacco control. Public health advocates also need to be willing to take prompt action to defend these programmes and hold public officials who attack tobacco control programmes accountable for their actions. PMID- 15333878 TI - Minors' tobacco possession law violations and intentions to smoke: implications for tobacco control. AB - OBJECTIVES: To test: (1) whether citation under the Minors in Possession (MIP) law, vicarious citation (knowing someone who was cited), and threat of driving licence suspension are associated with decreased intentions to smoke next year; and (2) whether the policy is differentially enforced. SUBJECTS: 28,249 white, Hispanic, and African American students in grades 6-12 (11-18 years old) participated in the study. METHOD: The 86 item anonymous Texas Youth Tobacco Survey was completed by students attending 37 schools in 14 east and central Texas communities. RESULTS: Hierarchical linear modelling showed that MIP citation was unrelated to the future smoking intentions of most youth. However, there was a negative association between citation and smoking intentions for ever daily smoking youth at four schools. Threat of licence suspension was associated with a lower likelihood of future smoking intentions among ever daily smoking youth and vicarious citation did not deter youth from future smoking. African American and Hispanic youth had a higher probability of being cited than their peers. CONCLUSIONS: Threat of driving licence suspension has the intended effect upon youth who are/were committed smokers and MIP citation has the intended effect upon committed smokers at only four schools. However, differential enforcement of the law based on ethnicity may be occurring. Before drawing firm conclusions, current findings must be replicated with longitudinal data to determine the consequences of citation on subsequent tobacco use. PMID- 15333879 TI - Current smoking and the risk of non-fatal myocardial infarction in the WHO MONICA Project populations. AB - BACKGROUND: Cohort studies have shown that smoking has a substantial influence on coronary heart disease mortality in young people. Population based data on non fatal events have been sparse, however. OBJECTIVE: To study the impact of smoking on the risk of non-fatal acute myocardial infarction (MI) in young middle age people. METHODS: From 1985 to 1994 all non-fatal MI events in the age group 35-64 were registered in men and women in the WHO MONICA (multinational monitoring of trends and determinants in cardiovascular disease) project populations (18,762 events in men and 4047 in women from 32 populations from 21 countries). In the same populations and age groups 65,741 men and 66,717 women participated in the surveys of risk factors (overall response rate 72%). The relative risk of non fatal MI for current smokers was compared with non-smokers, by sex and five year age group. RESULTS: The prevalence of smoking in people aged 35-39 years who experienced non-fatal MI events was 81% in men and 77% in women. It declined with increasing age to 45% in men aged 60-64 years and 36% in women, respectively. In the 35-39 years age group the relative risk of non-fatal MI for smokers was 4.9 (95% confidence interval (CI) 3.9 to 6.1) in men and 5.3 (95% CI 3.2 to 8.7) in women, and the population attributable fractions were 65% and 55%, respectively. CONCLUSIONS: During the study period more than half of the non-fatal MIs occurring in young middle age people can be attributed to smoking. PMID- 15333880 TI - Short term patterns of early smoking acquisition. AB - OBJECTIVE: To describe short term patterns of smoking acquisition exhibited by adolescent smokers. DESIGN: Interview records from the prospective development and assessment of nicotine dependence in youth study were examined retrospectively. Interviews were conducted three times per year over 30 months. SUBJECTS: 164 students in grades 7-9 (ages 12-15 years, 86 girls, 78 boys) who had used cigarettes at least twice. MAIN OUTCOME MEASURES: A continuous timeline of smoking activity, beginning with the subject's first use of tobacco and continuing through follow up, was translated into six patterns--abstinent, sporadic, occasional, daily, escalating, and intermittent. Outcome measures were the proportion of subjects starting/ending in each pattern, and the number of transitions per subject between patterns. RESULTS: There was a general but discontinuous progression from infrequent to more frequent use, with many interspersed periods of not smoking. Escalation to daily smoking was common after the development of dependence symptoms, but was rare among those who did not have symptoms. After the appearance of symptoms, both transitions to heavier daily smoking and attempts at cessation increased. CONCLUSIONS: Movement to heavier, more frequent smoking is generally unidirectional, although many youths attempt to quit one or more times. The appearance of any symptom of dependence altered the subsequent pattern of smoking behaviour. Future investigators might consider using more frequent data points and a continuous timeline to track smoking behaviour. PMID- 15333881 TI - Recent trends in home and work smoking bans. AB - OBJECTIVES: Home and work smoking bans at the national and state level in the USA, and their relation to smoking prevalence and to tobacco control policies, are examined. DATA: The Current Population Survey's 1992/93 and 1998/99 tobacco use supplement surveys are the primary data source, supplemented with information on state level tobacco control policies. METHODS: The national and state rate of bans are estimated, and changes over the course of the 1990s and their relation to smoking rates and tobacco control policies are examined. RESULTS: The prevalence of work and home bans has increased considerably between 1992/93 and 1998/99. By 1999, over 65% of the population age 15 and above work in places with smoking bans, and over 60% live in homes with bans. We found that states with lower than average ban rates in 1993 tended to have had larger increases in ban rates between 1993 and 1999. We also found that home and work bans became more prevalent in states with initially low smoking rates, and that the growth in home bans coincided with a declining prevalence of smoking. States with higher levels of bans by 1999 also tended to have higher cigarette taxes, stricter clean air laws, and media/comprehensive campaigns in place. CONCLUSIONS: The results indicate that lower smoking rates are associated with higher rates of work and home bans, although substantial progress has also been made by those states with initially low rates of bans. While further work is needed to establish the direction of causality, it would appear that reductions in smoking rates, either through stronger tobacco control policies or otherwise, may reduce exposure to tobacco smoke not only by reducing the number of smokers, but also through increasing the number of firms and homes with smoking restrictions. PMID- 15333882 TI - The economic burden of smoking in California. AB - OBJECTIVE: To develop estimates of the direct and indirect costs of smoking for California in 1999. METHODS: A prevalence based approach was used to estimate the annual costs of smoking. Econometric models were used to estimate the smoking attributable fraction (SAF) for direct costs (hospitalisations, ambulatory care, prescription drugs, home health care, and nursing home services) and indirect costs due to lost productivity from smoking related illness. The models controlled for socioeconomic factors and other risk behaviours. Epidemiological methods were used to estimate the SAF for indirect costs due to lost productivity from premature deaths. The SAFs were applied to total health care expenditures, days lost, and deaths to obtain smoking attributable total costs. RESULTS: In 1999, the total costs of smoking in California were 15.9 billion dollars, 475 dollars per resident, and 3331 dollars per smoker. Direct costs were 8.6 billion dollars (54% of the total), indirect costs due to lost productivity from illness were 1.5 billion dollars (10%), and indirect costs due to premature deaths were 5.7 billion dollars (36%). The cost of smoking was 9.4 billion dollars for men and 6.3 billion dollars for women. There were 43,137 deaths attributed to smoking, representing a total of 535,000 years of life lost. The value of life lost per death averaged 132,000 dollars, or 12.4 years. CONCLUSIONS: California smoking related costs are high. The cost methodology presented is useful for other states and nations interested in estimating their costs of smoking. Cost estimates can be used to evaluate the level of cigarette taxes and other policies related to smoking. PMID- 15333883 TI - Hedging their bets: tobacco and gambling industries work against smoke-free policies. AB - OBJECTIVE: To describe and understand the relationship between the tobacco and gambling industries in connection to their collaborative efforts to prevent smoke free casinos and gambling facilities and fight smoke-free policies generally. METHODS: Analysis of tobacco industry documents available online (accessed between February and December 2003). RESULTS: The tobacco industry has worked to convince the gambling industry to fight against smoke-free environments. Representatives of the gambling industry with ties to the tobacco industry oppose smoke-free workplaces by claiming that smoke-free environments hurt gambling revenue and by promoting ventilation as a solution to secondhand smoke. With help from the tobacco industry, the gambling industry has become a force at the American Society of Heating Refrigeration and Air Conditioning Engineers opposing smoke-free ventilation standards for the hospitality industry. CONCLUSION: Tobacco industry strategies to mobilise the gambling industry to oppose smoke free environments are consistent with past strategies to co-opt the hospitality industry and with strategies to influence policy from behind the scenes. Tobacco control advocates need to be aware of the connections between the tobacco and gambling industries in relation to smoke-free environments and work to expose them to the public and to policy makers. PMID- 15333884 TI - Out of the Smokescreen: does an anti-smoking advertisement affect young women's perception of smoking in movies and their intention to smoke? AB - OBJECTIVE: To evaluate the effect of an anti-smoking advertisement on young women's perceptions of smoking in movies and their intention to smoke. SUBJECTS/ SETTING: 2038 females aged 12-17 years attending cinemas in New South Wales, Australia. DESIGN/ INTERVENTION: Quasi-experimental study of patrons, who were surveyed after having viewed a movie at their local cinema. The control group was surveyed during week 1 and the intervention group, during week 2. Before seeing the movie in week 2, a 30 second anti-smoking advertisement was shown, which featured a well known female actor drawing attention to the prevalence of smoking in movies. OUTCOMES: Attitude of current smokers and non-smokers to smoking in the movies; intention of current smokers and non-smokers to be smoking in 12 months time. RESULTS: Among non-smokers, 48.2% of the intervention subjects thought that the smoking in the movie they viewed was "not OK" compared with 28.3% of the control subjects (p < 0.0001). However, there was no difference among smokers in the intervention (26.4%) and control (16.9%) groups (p = 0.28). A higher percentage of current smokers in the intervention group indicated they were unlikely to smoke in 12 months time (47.8%) than smokers in the control condition (31.9%) (p = 0.03). For non-smokers, there was no difference in smoking intentions between conditions, with 95% saying they would be unlikely to be smoking in 12 months time. CONCLUSIONS: This "real world" study suggests that placing an anti-smoking advertisement before movies containing smoking scenes can help to "immunise" young women against the influences of film stars smoking. PMID- 15333885 TI - The impact of anti-tobacco industry prevention messages in tobacco producing regions: evidence from the US truth campaign. AB - BACKGROUND: Adolescents who live in tobacco producing regions may not respond favourably to anti-industry ads. OBJECTIVE: To examine whether state level involvement in tobacco production appears to limit the effectiveness of anti industry ads to prevent tobacco use among adolescents in the USA. DESIGN: Time trend analyses were done using repeated cross sectional data from six waves of the Legacy Media Tracking Survey, which were collected between 1999 and 2003. SETTING AND PARTICIPANTS: 28,307 adolescents, ages 12-17 years, were classified as living in: tobacco producing states (TPS) (n = 1929); non-tobacco producing states (non-TPS) with low tobacco control funding comparable to TPS (n = 5323); non-TPS with relatively high funding (n = 15,076); and non-TPS with established anti-industry ad campaigns (n = 5979). MAIN OUTCOME MEASURES: Reactions to anti industry ads; strength of anti-industry attitudes/beliefs; changes in anti industry attitudes/beliefs over time. RESULTS: Ad reactions did not differ by state type. Multivariate adjusted time trend analyses indicated significant, comparable increases in anti-industry attitudes/beliefs since the onset of the truth campaign, in both TPS and non-TPS. Mediation analyses indicated that these increases were due, in part, to campaign exposure. CONCLUSIONS: Adolescents who live in tobacco producing regions appear to be as responsive to anti-industry ads as their counterparts in non-tobacco producing regions. This study provides further evidence for the effectiveness of such ads. PMID- 15333886 TI - Psychosocial work conditions, social capital, and daily smoking: a population based study. AB - OBJECTIVE: To investigate the associations between psychosocial conditions at work, social capital/social participation, and daily smoking. DESIGN/SETTING/PARTICIPANTS/MEASUREMENTS: The 2000 public health survey in Scania is a cross sectional postal questionnaire study with a 59% participation rate. A total of 5180 persons aged 18-64 years that belonged to the work force and the unemployed were included in this study. Logistic regression models were used to investigate the associations between psychosocial factors at work/unemployment, social participation, and daily smoking. Psychosocial conditions at work were defined according to the Karasek-Theorell demand-control/decision latitudes into relaxed, active, passive, and jobstrain categories. The multivariate analyses included age, country of origin, education and economic stress. RESULTS: 17.2% proportion of all men and 21.9% of all women were daily smokers. The jobstrain (high demands/low control) and unemployed categories had significantly higher odds ratios of daily smoking among both men and women compared to the relaxed (low demands/high control) reference category. The passive (low demands/low control), jobstrain, and unemployed categories were also significantly associated with low social participation. Low social participation was significantly and positively associated with daily smoking within each of the psychosocial work conditions and unemployed categories. CONCLUSIONS: The positive association between low social capital/low social participation and daily smoking is well known. However, both social participation and daily smoking are associated with psychosocial work conditions and unemployment. Psychosocial work conditions and unemployment may affect daily smoking both directly and through a pathway including social participation. PMID- 15333887 TI - Prevalence and correlates of internet cigarette purchasing among adult smokers in New Jersey. AB - OBJECTIVE: To examine the prevalence and correlates of internet cigarette purchasing among adult smokers. DESIGN: Analysis of internet purchasing in data from a population based telephone survey of New Jersey households. Logistic regression was used to determine factors associated with internet cigarette purchasing, adjusting for year, demographic, and smoking behaviour variables. PARTICIPANTS: 3447 current cigarette smokers pooled from three cross sectional surveys conducted in 2000, 2001, and 2002. MAIN OUTCOME MEASURES: Ever purchasing tobacco and usually buying cigarettes via the internet. RESULTS: Among all current cigarette smokers, ever having purchased tobacco via the internet increased from 1.1% in 2000 to 6.7% in 2002 and usually buying cigarettes via the internet increased from 0.8% in 2000 to 3.1% in 2002. Among current cigarette smokers with internet access, ever having purchased tobacco via the internet was higher among those who reported smoking 31 or more cigarettes per day (adjusted odds ratio (OR) 3.9, 95% confidence interval (CI) 1.5 to 10.2) and those without a past year quit attempt (adjusted OR 1.8, 95% CI 1.1 to 3.0). Usually purchasing cigarettes via the internet was higher among those aged 45-64 years (adjusted OR 4.4, 95% CI 1.1 to 17.1) and who reported having their first cigarette < or = 30 minutes after waking (adjusted OR 3.3, 95% CI 1.2 to 9.2). CONCLUSIONS: Although higher prices are known to reduce the demand for cigarettes, internet cigarette purchasing is likely to weaken this effect, particularly among heavy, more dependent smokers who are less interested in quitting. PMID- 15333888 TI - Current smoking among young adolescents: assessing school based contextual norms. AB - OBJECTIVE: To extend research on the relation of school based contextual norms to current smoking among adolescents by using three analytic techniques to test for contextual effects. It was hypothesised that significant contextual effects would be found in all three models, but that the strength of these effects would vary by the statistical rigor of the model. DESIGN: Three separate analytic approaches were conducted on baseline self report student survey data from a larger study to test the relation between school level perceived peer tobacco use and individual current smoking status. PARTICIPANTS: A representative sample of 5399 sixth through eighth grade students in 14 midwestern middle schools completed the survey. All enrolled sixth through eighth grade students were eligible to participate in the survey. The student participation rate was 91.4% for the entire sample, and did not differ significantly between the schools (range 82 100%). MAIN OUTCOME MEASURE: Thirty day cigarette smoking prevalence. RESULTS: A level 2 only model based on aggregated individual responses indicated that students in schools with higher average reported peer tobacco use were more likely to be current smokers than students in schools with lower average peer tobacco use. Using a level 1 only model based on individual responses indicated that the effect of school level perceived peer tobacco use on current smoking was significant when individual perceived peer tobacco use was excluded from the model but was non-significant when individual perceived peer tobacco use was added to the model. A multilevel model also indicated that the effect of school level perceived peer tobacco use on current smoking was not significant when individual perceived peer tobacco use was added to the model. CONCLUSION: The analytic approach used to examine contextual effects using individuals' reports of peer tobacco use norms that were aggregated to obtain a context measure of the school norms may produce statistical artefacts that distort the association of the school context in general, and peer tobacco use norms in particular, with increased risk for current smoking beyond the risk associated with individual factors. PMID- 15333889 TI - Stereotyping the smoker: adolescents' appraisals of smokers in film. AB - OBJECTIVE: To assess the relation between demographic factors and film smoking stereotypes in adolescents and the potential influence of smoker stereotypes on smoking susceptibility. DESIGN: A cross sectional questionnaire survey of school students (n = 3041) aged 12-13 and 16-17 years who were asked to describe the personal characteristics of female and male smokers in films. SETTING: 15 primary or intermediate schools and 10 secondary schools in Auckland, New Zealand. RESULTS: Appraisals of smokers in film were strongly influenced by age and sex with younger adolescents and males more likely to see female smokers as sexy, intelligent and healthy whereas older students and females more often appraised female smokers as stressed bored and depressed. Overall, image stereotypes (sexy, stylish) were more likely to be significantly associated with smoking susceptibility than emotional sensitivity stereotypes (stressed, depressed etc). CONCLUSIONS: Adolescents differ significantly in their appraisal of smokers in films; however, image based stereotypes, rather than emotional sensitivity stereotypes, are significantly associated with smoking susceptibility. PMID- 15333890 TI - Reaching youth at the point of sale: cigarette marketing is more prevalent in stores where adolescents shop frequently. AB - OBJECTIVE: Although numerous studies describe the quantity and nature of tobacco marketing in stores, fewer studies examine the industry's attempts to reach youth at the point of sale. This study examines whether cigarette marketing is more prevalent in stores where adolescents shop frequently. DESIGN, SETTING, AND PARTICIPANTS: Trained coders counted cigarette ads, products, and other marketing materials in a census of stores that sell tobacco in Tracy, California (n = 50). A combination of data from focus groups and in-class surveys of middle school students (n = 2125) determined which of the stores adolescents visited most frequently. MAIN OUTCOME MEASURES: Amount of marketing materials and shelf space measured separately for the three cigarette brands most popular with adolescent smokers and for other brands combined. RESULTS: Compared to other stores in the same community, stores where adolescents shopped frequently contained almost three times more marketing materials for Marlboro, Camel, and Newport, and significantly more shelf space devoted to these brands. CONCLUSIONS: Regardless of whether tobacco companies intentionally target youth at the point of sale, these findings underscore the importance of strategies to reduce the quantity and impact of cigarette marketing materials in this venue. PMID- 15333891 TI - Transfer of particulate matter pollution from smoking to non-smoking coaches: the explanation for the smoking ban on Italian trains. PMID- 15333892 TI - Deaths caused by secondhand smoke: estimates are consistent. PMID- 15333894 TI - Synthesis of DNA using a new two-step cycle. AB - For the first time a new, two-step method is described for synthesizing deoxyribonucleic acid. The approach uses 5'-carbonate protected 2' deoxynucleoside-3'-phosphoramidites as synthons and a peroxy anion buffer that removes the carbonate protecting group and oxidizes the internucleotide linkage. Following synthesis via this two-step cycle, oligomers are isolated by standard procedures. PMID- 15333895 TI - RNA synthesis using 2'-O-(tert-butyldimethylsilyl) protection. AB - This chapter enables the reader to carry out the solid-phase synthesis of ribonucleic acid (RNA) using beta-cyanoethyl phosphoramidite chemistry combined with tert-butyldimethylsilyl protection of the ribose 2'-hydroxyl group. Phosphoramidite monomers are activated with 5-benzylmercapto-1H-tetrazole enabling fast and highly efficient coupling to the 5'-hydroxyl group of the support-bound oligonucleotide. On completion of the synthesis, the stepwise deprotection of the nucleobase, phosphate, and ribose protecting groups is carried out using optimized protocols. Subsequently the various high-pressure (performance) liquid chromatography (HPLC) procedures are described enabling the purification and analysis of the RNA. For this purpose anion-exchange and reversed-phase HPLC are used singly or in combination according to the final purity requirement of the RNA. PMID- 15333896 TI - RNA oligonucleotide synthesis via 5'-silyl-2'-orthoester chemistry. AB - Rapid, reliable, and cost-efficient methods of ribonucleic acid (RNA) oligonucleotide synthesis are in demand owing to an increasing awareness of critical structural, functional, and regulatory roles of RNA throughout biology. The most promising area of growth and development is in RNA interference as an emerging technology for facilitating research in drug discovery and therapeutic intervention. Traditional methods of RNA synthesis, which are based on 2'-silyl protection strategies derived from deoxyribonucleic acid (DNA) synthesis strategies, are limited in their ability to produce oligos of sufficient purity and length for high-throughput applications. The more recently developed 5'-silyl 2'-acetoxy ethyl orthoester chemistry (2'-ACE trade mark ), circumvents several limitations of the 2'-silyl approaches. A clear improvement in RNA synthesis technology, 2'-ACE results in faster coupling rates, higher yields, greater purity, and superior ease of handling. Another advantage of the 2'-ACE protecting group strategy is that the molecules can be produced in an intermediately protected form that is soluble in aqueous solutions but resistant to nuclease attack. The chemistry can be scaled up or down and is flexible enough to allow for the incorporation of modifying groups if desired. A detailed description of the 2'-ACE protocol and procedures for end product analysis are presented. PMID- 15333897 TI - Dimethylthiarum disulfide: new sulfur transfer reagent in phosphorothioates oligonucleotide synthesis. AB - Dimethylthiarum disulfide (DTD) has been developed as a new and efficient sulfur transfer reagent for automated synthesis of phosphorothioate oligonucleotides using phosphoramidite chemistry. The traditional four-step automated oligonucleotide synthesis has been compressed to three-step protocol using DTD. This improvement allowed an overall 20% reduction in the solvent consumption and reduced the total synthesis time by 25%. The large-scale application of DTD has been successfully demonstrated by synthesis of therapeutically useful 20-mer phosphorothioate antisense oligonucleotides with excellent yield and purity. PMID- 15333898 TI - Assay for evaluating ribonuclease H-mediated degradation of RNA-antisense oligonucleotide duplexes. AB - Ribonucleases H are complex enzymes whose functions are not clearly understood, further compounded by the fact that multiple forms of the enzyme are present in various organisms. They are known to recognize and degrade the ribonucleic acid (RNA) strand of numerous deoxyribonucleic acid (DNA)-RNA duplex substrates, and so may provide a unique mode of therapeutic intervention at the genetic level of virtually any disease. We have therefore set out detailed procedures for conducting routine assays with almost any one of this family of enzymes by a straightforward assay aimed at identifying novel enzyme-activating antisense oligonucleotides (AONs). The procedures described herein should enable easy identification of potent AON molecules, provided that the RNA is appropriately labeled for subsequent visualization following the guidelines set forth in this protocol. PMID- 15333899 TI - Di- and oligonucleotide synthesis using H-phosphonate chemistry. AB - In this chapter, a concise account of the synthesis of oligonucleotides using the H-phosphonate methodology is given. It includes various methods for the preparation of the starting material, nucleoside 3'-H-phosphonate monoesters, their conversion into dinucleoside H-phosphonate diesters, oxidative transformations of dinucleoside H-phosphonates into the corresponding phosphate and phosphorothioate derivatives, and protocols for the synthesis of oligonucleotides and their phosphorothio analogs. PMID- 15333900 TI - Solid-phase synthesis of circular oligonucleotides. AB - A protocol for the straightforward preparation of small circular oligodeoxyribonucleotides (2-28 nt) is reported. The assembly of the oligonucleotide chain (standard phosphoramidite chemistry) and cyclization by the phosphotriester method take place on a tailor-made nucleotide-derivatized solid support. Although cyclization yields are moderate, the procedure exploits a synthesis design that allows selective cleavage of the circular oligonucleotide from the support, which facilitates isolation of the target molecule by simple filtration. PMID- 15333901 TI - Locked nucleic acid synthesis. AB - Methods and protocols for automated synthesis and purification of locked nucleic acid (LNA), a class of oligonucleotides obeying the Watson-Crick base-pairing rules but displaying unprecedented binding affinities toward complementary deoxyribonucleic acid (cDNA) and ribonucleic acid (RNA), is described. LNA and LNA-DNA chimeras containing phosphordiester or phosphorothioate linkages, or a mixture thereof, can be assembled by standard DNA synthesizers using 2-cyanoethyl DNA phosphoramidites and 2-cyanoethyl LNA phosphoramidites. Compared to the standard protocols used for DNA synthesis, slightly longer coupling time and oxidation time are needed for efficient oligomerization of LNA phosphoramidites. When the LNA has been assembled, it is removed from the solid support as the 5' end-O-dimethoxytrityl (DMT) protected LNA oligomer by treatment with concentrated aqueous ammonia that also removes the phosphate and nucleobase protecting groups. The crude DMT protected LNA product can be purified using, for example, reversed phase chromatography. PMID- 15333902 TI - Synthesis of DNA mimics representing HypNA-pPNA hetero-oligomers. AB - The methods for the synthesis and purification of negatively charged peptide nucleic acid (PNA)-relative deoxyribonucleic acid (DNA) mimics containing alternating residues of phosphono peptide nucleic acid (pPNA) monomers and PNA like monomers on the base of trans-4-hydroxy-L-proline are described. Examples of the chimeric oligomers hybridization with complementary DNA and ribonucleic acid fragments are demonstrated. PMID- 15333903 TI - Base-modified oligonucleotides with increased duplex stability: pyrazolo[3,4-d] pyrimidines replacing purines. AB - Oligonucleotides incorporating 8-aza-7-dazapurines (pyrazolo[3,4-d]pyrimidines) were synthesized. The corresponding nucleosides were prepared and were converted into phosphoramidites. The oligonucleotide duplex stability was studied and was compared to that of the parent compounds containing the canonical purine nucleosides. The presence of 7-halogeno or 7-alkynyl substituents increases the duplex stability significantly. PMID- 15333904 TI - Introduction of hypermodified nucleotides in RNA. AB - The anticodon domain of lysine transfer ribonucleic acid (tRNA) is a model system for investigation of the structural and biochemical effects of nucleoside posttranscriptional modification. To enable detailed study of the biophysical and structural effects of hypermodified nucleosides, methods have been developed to synthesize RNA oligonucleotides containing the modified nucleosides found in lysine tRNA. We describe in detail the synthesis of protected phosphoramidites of the nucleosides methylaminomethyl-2-thiouridine (mnm5s2U), methylcarboxymethyl-2 thiouridine (mcm5s2U), and 2-thiomethyl-N-6-carbamoylthreonyl-adenosine (ms2t6A). We also describe methods for using these nucleoside phosphoramidite reagents to synthesize RNA oligonucleotides with modified nucleosides incorporated at the specific sequence locations corresponding to their positions in the native lysine tRNAs. PMID- 15333905 TI - Chemical methods for peptide-oligonucleotide conjugate synthesis. AB - Methods of peptide-oligonucleotide conjugate synthesis are presented that may be useful in the study of cell targeting and delivery of oligonucleotides and their analogs. The first method involves total stepwise solid-phase synthesis on a single support. The second involves preparation of oligonucleotides containing 2' aldehydes and subsequent chemoselective ligation with functionalized peptides to form hydrazine, oxime, or thiazolidine linkages. PMID- 15333906 TI - Biotin-labeled oligonucleotides with extraordinarily long tethering arms. AB - We describe synthesis of four novel biotin phosphoramidites with tethering arms ranging from 20 to 74 atoms in length. One of these phosphoramidites is a uridine derivative with a biotin moiety attached through the 2'-position. The biotin phosphoramidites were synthetized based on robust and efficient methoxyoxalamido (MOX) and succinimido (SUC) precursor strategies from MOX/SUC precursors containing a secondary hydroxyl. They are highly stable in solution (coupling efficiency remains equally high for at least 2 wk after phosphoramidite installation on the synthesizer) and are ideal for the streamlined production of biotin-labeled oligonucleotides. Protocols for the synthesis of biotinylated primers and purification of biotinylated sequencing products by means of streptavidin-coated magnetic beads are also presented. PMID- 15333907 TI - Universal labeling chemistry for nucleic acid detection on DNA chips. AB - An efficient strategy for nucleic acid labeling and analysis on deoxyribonucleic acid (DNA) chips has been developed. This approach, which combines the fragmentation and the labeling steps, is based on the reactivity of the phosphates of DNA and ribonucleic acid (RNA) fragments and is using reporter molecules bearing a bromomethyl- or aryldiazomethane-reactive group. In this chapter, we describe the preparation of the reactive label and protocols for efficient labeling of any nucleic acid sequence, DNA or RNA, prior to their hybridization, detection, and analysis on DNA chips. PMID- 15333908 TI - Postsynthetic functionalization of triple helix-forming oligonucleotides. AB - The design of molecules that recognize specific sequence on the deoxyribonucleic acid (DNA) double helix would provide interesting tools to interfere with DNA information processing at an early stage of gene expression. This chapter describes in detail the protocol of conjugation between terminally phosphorylated oligonucleotides and chemically or biologically active ligands possessing electrophilic or nucleophilic functional groups. The synthetic procedure includes chemical activation of oligonucleotide terminal phosphate and introduction in this way of a nucleophilic or electrophilic group (such as amino or carboxyl groups) into oligonucleotide terminus using aliphatic amino group of a ligand or a linker. The attachment of a topoisomerase inhibitor camptothecin to a triple helix-forming oligonucleotide is taken as an example of such synthesis. The described method has general interest because any functional ligand containing a primary or secondary amino group or aliphatic carboxyl group could be attached to the terminal phosphate of an oligonucleotide in a similar way. PMID- 15333909 TI - Fluorescence-based on-line detection as an analytical tool in RNA electrophoresis. AB - In this chapter, we present methods for denaturing and native gel electrophoresis of nucleic acids based on fluorescently labeled probes and automatic signal detection by a deoxyribonucleic acid sequencer. Specific examples are given for the determination of ribonucleic acid (RNA) fragmentation patterns, of (deoxy)ribozyme kinetics, and of direct or competitive gel-shift assays. These methods can replace widely used radioisotope-based protocols, for example, for secondary structure mapping of RNA and for the characterization of nucleic acid ligand interactions. PMID- 15333910 TI - Design and optimization of molecular beacon real-time polymerase chain reaction assays. AB - During the last few years, several innovative technologies have become available for performing sensitive and accurate genetic analyses. These techniques use fluorescent detection strategies in combination with nucleic acid amplification protocols. Most commonly used is the real-time polymerase chain reaction (PCR). To achieve the maximum potential of a real-time PCR assay, several parameters must be evaluated and optimized independently. This chapter describes the different steps necessary for establishing a molecular beacon real-time PCR assay: (1) target design, (2) primer design, (3) optimization of the amplification reaction conditions using SYBR Green, (4) molecular beacon design, and (5) molecular beacon synthesis and characterization. The last section provides an example of a multiplex quantitative real-time PCR. PMID- 15333911 TI - High-throughput production of optimized primers (fimers) for whole-genome direct sequencing. AB - Fimers are specifically modified primers selected to inhibit nonspecific interactions occurring in cycle sequencing. They are postsynthetically derived from 2'-methoxyoxalamido or 2'-succinimido precursor oligonucleotides by treatment with appropriate small molecular weight modifiers (a primary aliphatic amine or hydroxide anion). We describe design, synthesis, and purification of fimers, and their use in protocols for direct sequencing of genomic deoxyribonucleic acid (DNA). Protocol for isolation of microbial genomic DNA is also reported. PMID- 15333912 TI - Nonenzymatic template-directed RNA synthesis. AB - The aim of this chapter is to provide reliable protocols for the study of nonenzymatic oligomerization reactions of ribonucleic acid and its analogs. Traditional radioactive labels are replaced by fluorescent dyes. Product analysis is either based on reversed-phase high-pressure (performance) liquid chromatography or on gel electrophoresis with on-line detection applying a deoxyribonucleic acid sequencer. Three examples of primer extensions are given, one of which demonstrates how the reaction of several primers may be monitored simultaneously. PMID- 15333913 TI - DNase I footprinting of small molecule binding sites on DNA. AB - Nuclease footprinting techniques were initially developed to investigate protein deoxyribonucleic acid (DNA) interactions but these tools of molecular biology have also become instrumental for probing sequence-selective binding of small molecules to DNA. Here, the method is described and technical details are given for performing deoxyribonuclease (DNase) I footprinting with DNA-binding drugs. An example is presented where DNase I is used (as well as DNase II and micrococcal nuclease) to probe the patterns of sequence-selective recognition of DNA by the anticancer antibiotic actinomycin D. DNase I is a convenient endonuclease for detecting and locating the position of actinomycin-binding sites within GC-rich sequences. PMID- 15333914 TI - Gene targeting using peptide nucleic acid. AB - A brief overview of the properties of the deoxyribonucleic acid mimic peptide nucleic acid (PNA) is given, and the recent progress in cellular delivery of PNA and of using PNA oligomers for antisense and antigene gene targeting is presented. PMID- 15333915 TI - Nucleic acid library construction using synthetic DNA constructs. AB - This chapter outlines seven synthetic and molecular biology techniques that allow the controlled synthesis of nucleic acid libraries. Specifically: (1) The high diversity chemical synthesis of point mutations; (2) the high-diversity chemical synthesis of point deletions; (3) the split-bead approach for constructing point mutation or deletion libraries with limited sequence diversity; (4) pool deprotection, gel purification, and quality-control techniques; (5) large-scale polymerase chain reaction amplification for the generation of high-diversity double-stranded deoxyribonucleic acid libraries; (6) type II restriction enzyme digestion techniques for the construction of long-sequence libraries containing minimal fixed sequence; and (7) extension techniques for the rapid synthesis of long, low-diversity oligonucleotide sequences. PMID- 15333916 TI - In vitro selection from combinatorial nucleic acid libraries. AB - Since the early 1990s, combinatorial deoxyribonucleic acid and ribonucleic acid libraries have been used to isolate specific ligands for a variety of target molecules, as well as nucleic acid-based catalysts for different reactions. These iterative procedures are based on the fact that nucleic acids can be enzymatically amplified. In this chapter, we describe the synthesis of such combinatorial libraries, their analysis, and basic procedures of in vitro selection. PMID- 15333917 TI - In vitro selection procedures for identifying DNA and RNA aptamers targeted to nucleic acids and proteins. AB - In vitro selection or systematic evolution of ligands by exponential enrichment is a combinatorial procedure that allows the identification of oligonucleotides showing properties of interest-so-called aptamers-through iterative selection/amplification rounds. Libraries containing as many as 1014 different sequences can be screened against a wide range of molecules. Ribonucleic acid (RNA), deoxyribonucleic acid (DNA), or chemically modified aptamers generally display high affinity and exquisite specificity of interaction with the target. Aptamers show a promising potential for diagnostic and therapeutic purposes. We describe here methods successfully used in our laboratory for the selection of RNA or DNA aptamers against an RNA structure (the transactivation response element of HIV-1) and a protein (the human ribonuclease H1). PMID- 15333918 TI - Short double-stranded ribonucleic acid as inhibitor of gene expression by the interference mechanism. AB - The rapid development of the small interfering ribonucleic acid (siRNA)-induced inhibition of the gene expression at the RNA level offers to research groups a new strategy for the understanding of gene functions. The siRNA approach is close to antisense oligonucleotide technology and takes advantage of the progress of chemically synthesized oligoribonucleotides. This approach for the mammalian cells was described by Elbashir et al. at the beginning of 2001, and in this chapter we describe methods for the design of siRNA molecules, solutions for efficiently transfecting cells, and methods for analyzing the inhibition of targeted genes. Methods for in vivo approach are also proposed. PMID- 15333920 TI - Structure of DsbC from Haemophilus influenzae. AB - Bacterial DsbC proteins are involved in rearranging or reducing mismatched disulfide bonds folding within the periplasm. The X-ray structure of the enzyme from Haemophilus influenzae has been solved and compared with the known structure of the Escherichia coli protein. The proteins act as V-shaped dimers with a large cleft to accommodate substrate proteins. The dimers are anchored by a small N terminal domain, but have a flexible linker region which allows the larger C terminal domain, with its reactive sulfhydryls, to clamp down on substrates. The overall folds are very similar, but the comparison shows a wider range of hinge motions than previously thought. The crystal packing of the H. influenzae protein allows the movement of the N-terminal domain with respect to the C-terminal domain through motions in the flexible hinge, generating high thermal parameters and unusually high anisotropy in the crystallographic data. PMID- 15333921 TI - Ab initio phasing based on topological restraints: automated determination of the space group and the number of molecules in the unit cell. AB - The connectivity-based phasing method has been demonstrated to be capable of finding molecular packing and envelopes even for difficult cases of structure determination, as well as of identifying, in favorable cases, secondary-structure elements of protein molecules in the crystal. This method uses a single set of structure factor magnitudes and general topological features of a crystallographic image of the macromolecule under study. This information is expressed through a number of parameters. Most of these parameters are easy to estimate, and the results of phasing are practically independent of these parameters when they are chosen within reasonable limits. By contrast, the correct choice for such parameters as the expected number of connected regions in the unit cell is sometimes ambiguous. To study these dependencies, numerous tests were performed with simulated data, experimental data and mixed data sets, where several reflections missed in the experiment were completed by computed data. This paper demonstrates that the procedure is able to control this choice automatically and helps in difficult cases to identify the correct number of molecules in the asymmetric unit. In addition, the procedure behaves abnormally if the space group is defined incorrectly and therefore may distinguish between the rotation and screw axes even when high-resolution data are not available. PMID- 15333922 TI - Water-molecule network and active-site flexibility of apo protein tyrosine phosphatase 1B. AB - Protein tyrosine phosphatase 1B (PTP1B) plays a key role as a negative regulator of insulin and leptin signalling and is therefore considered to be an important molecular target for the treatment of type 2 diabetes and obesity. Detailed structural information about the structure of PTP1B, including the conformation and flexibility of active-site residues as well as the water-molecule network, is a key issue in understanding ligand binding and enzyme kinetics and in structure based drug design. A 1.95 A apo PTP1B structure has been obtained, showing four highly coordinated water molecules in the active-site pocket of the enzyme; hence, the active site is highly solvated in the apo state. Three of the water molecules are located at positions that approximately correspond to the positions of the phosphate O atoms of the natural substrate phosphotyrosine and form a similar network of hydrogen bonds. The active-site WPD-loop was found to be in the closed conformation, in contrast to previous observations of wild-type PTPs in the apo state, in which the WPD-loop is open. The closed conformation is stabilized by a network of hydrogen bonds. These results provide new insights into and understanding of the active site of PTP1B and form a novel basis for structure-based inhibitor design. PMID- 15333923 TI - Accurate automatic protein models. AB - A method for automatic building of protein structures has been developed. The method is based on the concept of flexible structure units. A structure unit is a fragment (group) of about ten atoms that is positioned in the electron density by a phased rotation and translation function. The positions, orientations and internal torsion angles of all structure units are refined by a phased flexible refinement. Individual structure units are connected into polyalanine chains. The sequence is aligned by combined marker and rotamer methods. The side chains are built either by the marker method and a full conformational search or by the rotamer method. Side chains are independent structure units. The structure unit represents a generalized atom and the group model can be refined by the least squares method. The protein model is built with an accuracy of about 0.2 A at resolutions of 1.2-1.9 A. Partial results can be obtained at resolutions of between 2.0 and 2.3 A. PMID- 15333924 TI - Structure of apo acyl carrier protein and a proposal to engineer protein crystallization through metal ions. AB - A topic of current interest is engineering surface mutations in order to improve the success rate of protein crystallization. This report explores the possibility of using metal-ion-mediated crystal-packing interactions to facilitate rational design. Escherichia coli apo acyl carrier protein was chosen as a test case because of its high content of negatively charged carboxylates suitable for metal binding with moderate affinity. The protein was successfully crystallized in the presence of zinc ions. The crystal structure was determined to 1.1 A resolution with MAD phasing using anomalous signals from the co-crystallized Zn(2+) ions. The case study suggested an integrated strategy for crystallization and structure solution of proteins via engineering surface Asp and Glu mutants, crystallizing them in the presence of metal ions such as Zn(2+) and solving the structures using anomalous signals. PMID- 15333925 TI - Ligand-binding and metal-exchange crystallographic studies on shrimp alkaline phosphatase. AB - Alkaline phosphatases (APs) are homodimeric metalloenzymes that catalyze the hydrolysis and transphosphorylation of phosphate monoesters. Each monomer contains a metal-binding triad that for optimal activity is usually occupied by two zinc ions and one magnesium ion. The recently determined crystal structure of cold-active shrimp alkaline phosphatase (SAP) was, however, fully occupied by zinc ions. This paper describes a metal-exchange experiment in which the zinc ion in one binding site (referred to as the M3 site) is replaced by magnesium. Crystal structures revealed a concomitant structural change: the metal exchange causes movement of a ligating histidine into a conformation in which it does not coordinate to the metal ion. The M3 site is relevant to catalysis: its occupation by magnesium is postulated to favour catalysis and it has been suggested to be a regulatory site for other APs. Further crystallographic studies show that ligand binding can induce a conformational change of an active-site arginine from a 'non docked' (non-interacting) to a 'docked' conformation (interacting with the ligand). The first conformation has only been observed in SAP, while the latter is common in available AP structures. The observation that the arginine does not always bind the substrate may explain the increased catalytic efficiency that is generally observed for cold-active enzymes. PMID- 15333926 TI - Statistical experimental design of protein crystallization screening revisited. AB - A statistical experimental design approach was used to prepare a set of solutions for the screening of protein crystallization conditions. This approach is shown to be amenable to quantitative evaluation and therefore to the rational optimization of the screening results. All solutions contain a cryoprotectant, thus eliminating the need for subsequent optimization of crystal freezing conditions. PMID- 15333927 TI - Structure of the cytosolic Cu,Zn superoxide dismutase from Schistosoma mansoni. AB - Cu,Zn superoxide dismutase (Cu,Zn SOD) is an essential enzyme for protecting cells from the toxic effects of reactive oxygen species. In humans, two distinct Cu,Zn SOD genes are located on chromosomes 4 and 21 and mutations in the latter have been associated with familial amyotrophic lateral sclerosis. Similarly, schistosomes (trematode parasites responsible for the chronically debilitating disease schistosomiasis) also produce two distinct Cu,Zn SODs, in this case one cytosolic and one bearing a signal peptide. The crystal structure of the cytosolic form of the enzyme from the human trematode Schistosoma mansoni (SmCtSOD) was solved and refined to a resolution of 2.2 A (space group P2(1)2(1)2(1), R = 17.6% and R(free) = 24.1%) and 1.55 A (space group P2(1), R = 15.7% and R(free) = 17.1%). This is the first report of a crystal structure of a Cu,Zn superoxide dismutase derived from a human parasite. Alternate positions for the catalytic copper and its water ligand were refined for the 1.55 A SmCtSOD model, but the most interesting structural differences between SmCtSOD and the human homologue reside in the loops used for electrostatic guidance of the substrate to the enzyme active site. PMID- 15333928 TI - Observation of time-resolved structural changes by linear interpolation of highly redundant X-ray diffraction data. AB - A new experimental strategy is described for obtaining time-resolved protein structural changes using monochromatic X-ray crystallographic data. The method is based on time-dependent linear interpolation of observed intensity variations during conventional X-ray diffraction data collection. The method benefits from high data redundancy. Although the method was developed to examine time-dependent X-ray-induced crystal decay, it is potentially applicable to a variety of time dependent crystallographic studies, including structural determination of chemical intermediates in enzyme reactions and X-ray-induced unfolding of proteins with multiple disulfide bonds. PMID- 15333929 TI - A redetermination of the structure of the triple mutant (K53,56,120M) of phospholipase A2 at 1.6 A resolution using sulfur-SAS at 1.54 A wavelength. AB - The crystal structure of the triple mutant K53,56,120M of bovine pancreatic phospholipase A(2) has been redetermined using sulfur single-wavelength anomalous scattering. The synchrotron data were collected at lambda = 1.54 A and the crystal diffracted to 1.6 A resolution. The program SOLVE was used to locate the heavy atoms and to estimate the initial phases and the resulting map was then subjected to RESOLVE. The output of 455 non-H atoms, including 12 S atoms, one calcium ion and one chloride ion, were then subjected to ARP/wARP followed by REFMAC. With the improved phases, the automatic model building successfully built more than 85% of the 123 residues, excluding the N- and C-terminal residues. The final crystallographic R factor is 17.7% (R(free) = 21.7%). The refined model consists of 954 non-H protein atoms, 165 water O atoms, three 2-methyl-2,4 pentanediol (MPD) molecules, one calcium ion and one chloride ion. The present work is yet another example that shows the utility of single-wavelength anomalous scattering data for solving a protein structure. PMID- 15333930 TI - Structure of the Yersinia pestis type III secretion chaperone SycH in complex with a stable fragment of YscM2. AB - Pathogenic Yersinia species use a type III secretion system to inject cytotoxic effector proteins directly into the cytosol of mammalian cells, where they neutralize the innate immune response by interfering with the signal-transduction pathways that control phagocytosis and inflammation. To be exported efficiently, some effectors must transiently associate with cognate cytoplasmic secretion chaperones. SycH is the chaperone for YopH, a potent eukaryotic-like protein tyrosine phosphatase that is essential for virulence. SycH also binds two negative regulators of type III secretion, YscM1 and YscM2, both of which share significant sequence homology with the chaperone-binding domain of YopH. Here, the structure of a complex between SycH and a stable fragment of YscM2 that was designed on the basis of limited proteolysis experiments is presented. The overall fold of SycH is very similar to the structures of other homodimeric secretion chaperones that have been determined to date. YscM2 wraps around SycH in an extended fashion, with some secondary but no tertiary structure, assuming a conformation distinct from the globular fold that it is predicted to adopt in the absence of SycH. PMID- 15333931 TI - Structure of serine acetyltransferase from Haemophilus influenzae Rd. AB - The crystal structure of serine acetyltransferase (SAT) from Haemophilus influenzae Rd determined at 2.7 A resolution is presented. SAT is a member of a family of hexapeptide-containing transferases that contain six-residue tandem repeats (LIV)-G-X(4) that have been shown to form left-handed parallel beta helices. In the current structure, each protomer is comprised of two domains: an N-terminal alpha-helical domain and a C-terminal left-handed parallel beta-helix domain. Although other members of this protein family are known to form trimeric structures, SAT forms a dimer of trimers in which the trimer interface is mediated through interactions between both the beta-helix domains and N-terminal domains; these trimers dimerize through contacts in the N-terminal domain. All dimer-of-trimer interactions are mediated through amino acids within an N terminal extension common only to a subset of SATs, suggesting that members of this subfamily may also adopt hexameric structures. Putative active sites are formed by crevices between adjacent protomers in a trimer. Thus, six independent active sites exist in the hexameric enzyme complex. PMID- 15333932 TI - Structural genomics of Caenorhabditis elegans: structure of the BAG domain. AB - Binding of the BAG domain to the eukaryotic chaperone heat-shock protein (Hsp70) promotes ATP-dependent release of the protein substrate from Hsp70. Although the murine and human BAG domains have been shown to form an antiparallel three-helix bundle, the Caenorhabditis elegans BAG domain is formed by two antiparallel helices, while the third helix is extended away and stabilized by crystal-packing interactions. A small beta-sheet between helices 2 and 3 interferes with formation of the intramolecular three-helix bundle. However, intermolecular three helix bundles are observed throughout the crystal packing and suggest that stable functional dimers and tetramers can be formed in solution. The structure may represent a new folding type of the BAG domain. PMID- 15333933 TI - Cloning, expression, purification, crystallization and initial crystallographic analysis of transcription factor DksA from Escherichia coli. AB - The Escherichia coli gene encoding a regulator of stringent response and virulence, DksA, which contains a canonical Zn finger motif, was cloned and expressed, and the purified protein was crystallized by the hanging-drop vapor diffusion technique in two different space groups, P2(1)2(1)2(1) (a = 91.32, b = 96.59, c = 117.48 A) and C222 (a = 80.6, b = 115.1, c = 149.57 A). The crystals belonging to space group P2(1)2(1)2(1), improved by macroseeding, diffract beyond 2.0 A at a synchrotron. Three complete atomic resolution multiple anomalous dispersion diffraction data sets were collected from the same crystal of the P2(1)2(1)2(1) crystal form at the absorption edge for Zn atoms. PMID- 15333934 TI - Cloning, expression, crystallization and initial crystallographic analysis of the C-terminal domain of the amyloid precursor protein APP. AB - Alzheimer's disease is associated with typical brain deposits (senile plaques) consisting mainly of neurotoxic amyloid beta-peptides. These are proteolytically derived from the large type I transmembrane protein amyloid precursor protein (APP), which is possibly involved in signal transduction. The large C-terminal domain CAPPD of the human APP ectodomain has been cloned, expressed in large amounts in Escherichia coli and purified to homogeneity. Well diffracting tetragonal crystals have been obtained and native data have been collected to 2.1 A. Initial experimental phases from a three-wavelength MAD experiment using (NH(4))(2)OsCl(6)-derivatized crystals are of good quality and show mostly alpha helical conformations. PMID- 15333935 TI - Expression, purification, crystallization and preliminary X-ray diffraction studies of the cmcI component of Streptomyces clavuligerus 7alpha-cephem methoxylase. AB - Cephamycins are broad-spectrum beta-lactam antibiotics that show resistance to certain forms of beta-lactamases. They differ from cephalosporins by the presence of a methoxyl group at the C-7alpha position. The gene products of cmcI and cmcJ are believed to control 7alpha-methoxylation of cephalosporins through successive steps of hydroxylation and methylation. Here, the expression, purification, crystallization and initial data-collection statistics of the 236-amino-acid protein product of cmcI from Streptomyces clavuligerus is reported. The crystals belong to space group P2(1), with unit-cell parameters a = 93.6, b = 182.6, c = 103.2 A, beta = 91.05 degrees. Diffraction data were collected to 2.5 A. PMID- 15333936 TI - Crystallization and preliminary X-ray analysis of carboxypeptidase Y inhibitor IC complexed with the cognate proteinase. AB - Carboxypeptidase Y (CPY) inhibitor I(C) is a naturally occurring serine carboxypeptidase inhibitor from Saccharomyces cerevisiae, the sequence of which is not homologous with any other known proteinase inhibitor and is classified as the phosphatidylethanolamine-binding protein (PEBP). I(C) has been crystallized in complex with the deglycosylated form of CPY by the hanging-drop vapour diffusion technique with ammonium sulfate as a precipitant. The crystals of the complex belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 81.13, b = 186.6, c = 65.14 A. Diffraction data were collected to 2.7 A resolution. Structure determination of the complex is in progress by the molecular replacement method using the structure of CPY as a search model. PMID- 15333937 TI - Crystallization of a non-B and a B mutant HIV protease. AB - HIV polymorphism is responsible for the selection of variant viruses resistant to inhibitors used in AIDS treatment. Knowledge of the mechanism of resistance of those viruses is determinant to the development of new inhibitors able to stop, or at least slow down, the disease's progress caused by new mutations. In this paper, the crystallization and preliminary crystallographic structure solution for two multi-resistant 99 amino acid HIV proteases, both isolated from Brazilian patients failing intensive anti-AIDS therapy are presented, viz. the subtype B mutant, with mutations Q7K, S37N, R41K, K45R, I54V, L63P, A71V, V82A and L90M, and the subtype F (wild type), naturally carrying mutations Q7K, I15V, E35D, M36I, S37N, R41K, R57K, D60E, Q61N, I62V, L63S, I64L and L89M, with respect to the B consensus sequence. Both proteins crystallized as a complex with the inhibitor TL-3 in space group P6(1)22. X-ray diffraction data were collected from these crystals to resolutions of 2.1 and 2.6 A for the subtype B mutant and subtype F wild type, respectively, and the enzyme structures were solved by molecular replacement. The crystals of subtype F HIV protease are, to the best of the authors' knowledge, the first protein crystals obtained for a non-B HIV protease. PMID- 15333938 TI - Purification, crystallization and preliminary crystallographic data of the m3G cap-binding domain of human snRNP import factor snurportin 1. AB - The nuclear import of spliceosomal UsnRNPs is mediated by the transport adaptor snurportin 1 (SPN1), which specifically recognizes the 2,2,7-trimethylguanosine (m(3)G) cap at the 5' end of UsnRNAs. Human SPN1 was overexpressed as a GST fusion protein in Escherichia coli and purified to homogeneity. Since full-length SPN1 did not crystallize, limited proteolysis experiments were performed and stable digestion products were analyzed for functionality with respect to m(3)G cap-binding activity and subsequently used for crystallization trials. Well diffracting single crystals of a truncated SPN1 m(3)G cap-binding domain (residues 79-300) were obtained after two rounds of seeding. The crystals belong to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 57.47, c = 130.09 A, alpha = beta = gamma = 90 degrees. Crystals contain one molecule in the asymmetric unit and diffract to a resolution limit of 2.9 A. PMID- 15333939 TI - Biochemical, biophysical and preliminary X-ray crystallographic analyses of the fusion core of Sendai virus F protein. AB - It is emerging that enveloped viruses may adopt a unique entry/fusion mechanism; in paramyxoviruses, including Sendai virus (SeV), the attachment protein HN (or its homologue H or G) binds a cellular receptor which triggers conformational changes of its fusion protein, F. There are at least three conformations of the F protein in the current fusion model: the pre-fusion native conformation, the pre hairpin intermediate conformation and the post-fusion coiled-coil conformation. The fusion mechanism of SeV, a member of the Paramyxoviridae family, has been well established and several structural and functional domains or modules have been proposed from studies of its F protein. However, biochemical and biophysical studies of the heptad-repeat (HR) regions (HR1 and HR2) have not been systematically carried out. HR1 and HR2 strongly interact with each other to form a stable six-helix coiled-coil bundle as the post-fusion conformation. In this study, a single-chain HR1-linker-HR2 protein of SeV was prepared in an Escherichia coli expression system and biochemical and biophysical analyses showed it to form a typical six-helix coiled-coil bundle; its trigonal crystals diffracted X-rays to 2.5 A resolution. The crystal structure will help to reveal the structural requirements of the post-fusion coiled-coil conformation of SeV F protein. PMID- 15333940 TI - Crystallization and preliminary X-ray analyses of desulfurization enzyme DszB and its C27S mutant complexed with biphenyl-2-sulfinic acid. AB - DszB is a hydrolase involved in the biodegradation of dibenzothiophene in the soil bacterium Rhodococcus sp. IGTS8. DszB catalyzes the hydrolysis of 2' hydroxybiphenyl-2-sulfinic acid to sulfite and biphenyl-2-ol. DszB and DszB C27S mutant complexed with biphenyl-2-sulfinic acid were crystallized and preliminary X-ray crystallographic analyses were conducted. The crystals of DszB were found to belong to the orthorhombic P2(1)2(1)2(1) space group, with unit-cell parameters a = 36.7, b = 82.6, c = 139.6 A, and to contain one molecule of DszB in the asymmetric unit. Crystals of DszB C27S complexed with biphenyl-2-sulfinic acid belong to space group C2, with unit-cell parameters a = 153.4, b = 45.9, c = 112.9 A, beta = 115.93 degrees. The calculated Matthews coefficient V(M) for the C2 crystals is approximately 2.3 A(3) Da(-1) if two molecules of DszB are present in the asymmetric unit. PMID- 15333941 TI - Crystallization and preliminary X-ray diffraction analysis of HeLp, a heme lipoprotein from the hemolymph of the cattle tick Boophilus microplus. AB - The main protein present in the hemelymph of the cattle tick Boophilus microplus is a lipoprotein able to bind heme (HeLp). It has an apparent molecular weight of 354 000 Da and is composed of two polypeptide chains found in stoichiometric amounts. It contains 33% lipids. The protein was crystallized using the hanging drop vapour-diffusion method in the presence of 1,6-hexanediol as a precipitant. X-ray diffraction data were collected to 2.1 A resolution using a synchrotron radiation source. The crystal belongs to the triclinic space group P1, with unit cell parameters a = 90.58, b = 105.50, c = 116.14 A, alpha = 112.40, beta = 111.64, gamma = 91.35 degrees. Owing to the lack of information about the amino acid sequence, the structure of HeLp will be solved by the use of heavy atoms. Several possible derivatives have been collected and analysis is under way. PMID- 15333942 TI - Crystallization and preliminary X-ray analysis of Escherichia coli MutT in binary and ternary complex forms. AB - During replication, Escherichia coli MutT prevents the misincorporation of mutagenic 8-oxoguanine into nascent DNA strands opposite adenine by hydrolyzing 8 oxo-dGTP in nucleotide pools to 8-oxo-dGMP. E. coli MutT is the most widely investigated member of the Nudix hydrolase family, which is large and found in all organisms. By co-crystallization of MutT with 8-oxo-dGMP, a reaction product, crystals of the binary complex were obtained using ammonium sulfate as a precipitant. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 37.9, b = 56.0, c = 59.4 A. Assuming the presence of one protein nucleotide complex in the asymmetric unit, the Matthews coefficient V(M) is 2.1 A(3) Da(-1). Crystals of the ternary complex were prepared by soaking crystals of the binary complex in 1 mM MnCl(2) solution. They diffracted to 1.96 and 2.56 A resolution, respectively. PMID- 15333943 TI - Crystallization and preliminary X-ray analysis of heparinase II from Pedobacter heparinus. AB - Heparinase II from Pedobacter heparinus (formerly Flavobacterium heparinum), which acts on both heparin and heparan sulfate, is one of several glycosaminoglycan-degrading enzymes produced by this organism. This enzyme, with a molecular weight of 84 kDa, utilizes a lytic mechanism to cleave the alpha(1-4) glycosidic bond between hexosamine (D-glucosamine) and L-iduronic or D-glucuronic acid, resulting in a product with an unsaturated sugar ring at the non-reducing end. The enzyme was crystallized by the hanging-drop vapour-diffusion method. The crystals belong to orthorhombic space group P2(1)2(1)2(1) and diffract to 2 A resolution. There are two molecules in the asymmetric unit, consistent with the finding that recombinant heparinase II functions as a dimer in solution. PMID- 15333944 TI - Purification, crystallization and preliminary X-ray analysis of Triatoma virus (TrV) from Triatoma infestans. AB - Triatoma virus (TrV) is a viral pathogen of the blood-sucking reduviid bug Triatoma infestans, the most important vector of American human trypanosomiasis (Chagas' disease). TrV has been putatively classified as a member of the Cripavirus genus (type cricket paralysis virus) in the Dicistroviridae family. This work describes the purification of TrV particles from infected T. infestans and their crystallization and preliminary crystallographic analyses. Two different crystal forms, rhombohedral and orthorhombic, were obtained at room temperature by the hanging-drop vapour-diffusion technique using polyethylene glycol and polyethylene glycol monomethylether as precipitants. The rhombohedral crystals have unit-cell parameters a = b = 306.6, c = 788.4 A (hexagonal setting), diffract to 3.2 A resolution and contain one-third of the viral particle per asymmetric unit. The orthorhombic crystals have cell parameters a = 336, b = 351, c = 332 A, diffract to about 2.5 A resolution, and contain one-half of a virus particle in the asymmetric unit. A complete diffraction data set has been collected to 3.2 A resolution, using synchrotron radiation, from a single rhombohedral crystal under cryogenic conditions. PMID- 15333945 TI - Expression, purification and preliminary crystallographic studies of human coactosin-like protein. AB - Coactosin-like protein (CLP) is an actin-binding protein as well as a 5 lipoxygenase binding partner. Human coactosin-like protein has been expressed in high yield and the His-tagged protein was purified by affinity chromatography. Several different crystal forms were obtained by the hanging-drop vapour diffusion method. X-ray diffraction data to 2.0 A resolution were collected from the best crystal. The space group was determined to be P2(1)2(1)2(1), with unit cell parameters a = 38.4, b = 48.7, c = 72.6 A. PMID- 15333946 TI - Crystallization and preliminary crystallographic studies of the cofactor-binding domain of the LysR-type transcriptional regulator Cbl from Escherichia coli. AB - Cbl (CysB-like protein) is a member of the family of LysR-type transcriptional regulators (LTTRs) and controls genes engaged in sulfur assimilation in Escherichia coli. It has been postulated that adenosine 5-phosphosulfate (APS) is responsible for abolishing Cbl-activated transcription from the ssu promoter (Bykowski et al., 2002). To elucidate the structural basis of Cbl function and to confirm the role of APS as an anti-inducer, the cofactor-binding domain of Cbl (c Cbl, MW = 26 kDa) was cloned, purified and crystallized in the presence of APS. The crystals belong to space group C222(1), but show substantial variation of the unit-cell parameters and diffraction anisotropy. Despite this, X-ray data extending to 3.0 A resolution have been collected and solution of the structure by molecular replacement is in progress. PMID- 15333947 TI - Crystallization of the xeroderma pigmentosum group F endonuclease from Aeropyrum pernix. AB - The xeroderma pigmentosa group F protein (XPF) is a founding member of a family of 3'-flap endonucleases that play an essential role in nucleotide-excision repair, DNA replication and recombination. The XPF gene has been cloned from Aeropyrum pernix, encoding a 254-residue protein (apXPF). Recombinant protein was produced in Escherichia coli and purified by three chromatographic steps. Three different crystal forms of apXPF were grown in trigonal, monoclinic and triclinic systems. The trigonal crystals diffracted to 2.8 A and were grown in the presence of double-stranded DNA. Monoclinic crystals were grown without DNA and diffracted to 3.2 A. Triclinic crystals were grown from a truncated apXPF protein lacking the tandem helix-hairpin-helix motifs and diffracted to 2.1 A. PMID- 15333948 TI - Cloning, expression, purification, crystallization and preliminary X-ray crystallographic investigations of a unique editing domain from archaebacteria. AB - Threonyl-tRNA synthetase (ThrRS) faces a crucial double-discrimination problem during the translation of genetic code. Most ThrRSs from the archaeal kingdom possess a unique editing domain that differs from those of eubacteria and eukaryotes. In order to understand the structural basis of the editing mechanism in archaea, the editing module of ThrRS from Pyrococcus abyssi comprising of the first 183 amino-acid residues was cloned, expressed, purified and crystallized. The crystals belong to the trigonal space group P3(1(2))21, with one molecule in the asymmetric unit. PMID- 15333949 TI - Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum. AB - Conjugated bile salt hydrolase (BSH) catalyses the hydrolysis of the amide bond that conjugates bile acids to glycine and to taurine. The BSH enzyme from Bifidobacterium longum was overexpressed in Escherichia coli BL21(DE3), purified and crystallized. Crystallization conditions were screened using the hanging-drop vapour-diffusion method. Crystal growth, with two distinct morphologies, was optimal in experiments carried out at 303 K. The crystals belong to the hexagonal system, space group P622 with unit-cell parameters a = b = 124.86, c = 219.03 A, and the trigonal space group P321, with unit-cell parameters a = b = 125.24, c = 117.03 A. The crystals diffracted X-rays to 2.5 A spacing. Structure determination using the multiple isomorphous replacement method is in progress. PMID- 15333950 TI - Crystals of X29, a Xenopus laevis U8 snoRNA-binding protein with nuclear decapping activity. AB - Eukaryotic ribosome biosynthesis requires modification (methylation, pseudouridylation) and nucleolytic processing of precursor ribosomal RNAs in the nucleolus. The RNA components of the small nucleolar RNPs (snoRNAs) are essential for many of these events. One snoRNP, called U8, is necessary for maturation of 5.8S and 28S rRNA in vertebrates. In Xenopus laevis, U8 snoRNA was found to bind specifically and with high affinity to a protein called X29. X29 is a Nudix hydrolase, a nucleotide diphosphatase that removes the m(7)G and m(227)G caps from U8 and other RNAs. X29 requires an RNA as substrate and cap analogues are not substrates or inhibitors of cleavage. To study the determinants of X29 activity and its specificity for U8 RNA substrate, X29 was crystallized in an orthorhombic crystal form that diffracts to 2.1 A resolution. PMID- 15333951 TI - Optimum solubility (OS) screening: an efficient method to optimize buffer conditions for homogeneity and crystallization of proteins. AB - One of the most critical steps in the preparation of protein samples for structural studies by X-ray crystallography is to obtain biochemically pure and conformationally homogenous protein samples. Very often, the purified sample does not meet these qualifications and therefore does not crystallize. A screening method, Optimum Solubility Screen, has been developed that consists of two steps. The first step selects a better buffer than that used during purification. 24 different buffers ranging from pH 3 to pH 10 are screened using a vapor-diffusion method and very small amounts of protein. The solubility of the protein is first determined by visual examination using a light microscope and those drops that remain clear after 24 h are further evaluated using dynamic light scattering. If the results from the first step are still not satisfactory, a second step explores a variety of chemical additives in order to improve the monodispersity of the protein sample. In 64% of the cases, crystallization was successful from proteins that had initially shown high levels of aggregation. This screen can be configured to perform in an automated high-throughput mode and can be expanded for additional buffers and additives. PMID- 15333952 TI - A preliminary solubility screen used to improve crystallization trials: crystallization and preliminary X-ray structure determination of Aeropyrum pernix flap endonuclease-1. AB - Crystallization of protein and protein complexes is a multi-parametric problem that involves the investigation of a vast number of physical and chemical conditions. The buffers, salts and additives used to prepare the protein will be present in every crystallization condition. It is imperative that these conditions be defined prior to crystal screening since they will have a ubiquitous involvement in the crystal-growth experiments. This study involves the crystallization and preliminary analysis of the flap endonuclease-1 (FEN-1) DNA repair enzyme from the crenarchaeal organism Aeropyrum pernix (Ape). Ape FEN-1 protein in a standard chromatography buffer had only a modest solubility and minimal success in crystallization trials. Using an ion/pH solubility screen, it was possible to dramatically increase the maximum solubility of the protein. The solubility-optimized protein produced large diffraction-quality crystals under multiple conditions in which the non-optimized protein produced only precipitate. Only minor adjustments of the conditions were required to produce single diffraction-quality crystals. The native Ape FEN-1 crystals diffract to 1.4 A resolution and belong to space group P6(1), with unit-cell parameters a = b = 92.8, c = 80.9 A, alpha = beta = 90, gamma = 120 degrees. PMID- 15333953 TI - APRV - a program for automated data processing, refinement and visualization. AB - APRV (Automatic Processing, Refinement and Visualization) is a new program that enables high-throughput batch processing of crystallographic data. The program combines processing of raw diffraction images, initial structure refinement and visual inspection of resulting electron density into a seamless one-step procedure, during which all relevant parameters are refined automatically. It is controlled by a user-friendly graphical interface, facilitating operation by non experts. PMID- 15333954 TI - MAD phasing: choosing the most informative wavelength combination. AB - Two algorithms are described for limiting data resolution and for predicting the most informative wavelength combinations in MAD techniques. Both have been successfully tested using experimental data from a large set of test structures. PMID- 15333955 TI - Structure of D-ribulose 5-phosphate 3-epimerase from Synechocystis to 1.6 A resolution. AB - The crystal structure of D-ribulose 5-phosphate 3-epimerase (RPE) from the cyanobacterium Synechocystis was determined by X-ray crystallography to 1.6 A resolution. The enzyme, which catalyzes the epimerization of D-ribulose 5 phosphate and D-xylulose 5-phosphate, assembles as a hexamer of (beta/alpha)(8) barrels in the crystallographic asymmetric unit. The active site is highly similar to those of two previously reported RPEs and provides further evidence for essential catalytic roles for several active-site residues. PMID- 15333956 TI - CLIMS: crystallography laboratory information management system. AB - Macromolecular crystallography requires simple yet effective means of organizing and managing the large amounts of data generated by crystallization experiments. There are several freely available web-based Laboratory Information Management Systems (LIMS) that assist in these tasks. These, however, rely on the limited user interfaces allowed in HTML-based web pages. To address this limitation, a new LIMS for protein crystallization, which features a novel rich graphical user interface (GUI) to a relational database, has been developed. This application, which is called CLIMS (Crystallography LIMS), assists in all aspects of protein crystallization projects: protein expression, handling, crystallization optimization, visualization of results and preliminary diffraction data. Extensive use of templates, particularly for commercial screens and common optimization grid screens, exploits the redundancy in experimental setups. The crystallization tray is the central focus of the graphical interface, thus facilitating rapid visualization and annotation of results. CLIMS was developed specifically to cater for the needs of individual laboratories requiring an intuitive and robust system for managing crystallization experiments and is freely available. PMID- 15333959 TI - How technology has transformed the one-visit initial exam. PMID- 15333960 TI - Tooth movement with Essix mounding. PMID- 15333961 TI - Debonding a new ceramic bracket: a clinical study. PMID- 15333962 TI - Piezoelectric surgery for exposure of palatally impacted canines. PMID- 15333964 TI - First molar bypass technique for uprighting ectopic second molars. PMID- 15333965 TI - Treatment of hyperlipidemia in nonalcoholic fatty liver disease: fat for thought. PMID- 15333966 TI - Serum hyaluronic acid concentrations in children with cirrhosis. AB - BACKGROUND: Liver disease is associated with increased levels of hyaluronic acid (HA). AIM: To evaluate serum HA concentrations in children with cirrhosis and its relation with liver function tests and Child-Pugh score. METHODS: Twenty-two children with biopsy-proven liver cirrhosis were studied. All were assessed for the presence of ascites or encephalopathy and liver function tests were performed. Patients were categorized according to Child-Pugh criteria. Serum HA was measured using microELISA (normal 0-100 ng/mL). Twenty-two children with chronic hepatitis B and no cirrhosis were studied as controls. RESULTS: Serum HA level in the cirrhotic children was 85.2 (72.8) ng/mL; levels were high (166.0 [46.3] ng/mL; range 115-246) in 8 (36.4%) patients. Three of 11 (27.2%) Child Pugh class A patients, 3 of 8 (37.5%) class B patients, and 2 of 3 (66.7%) class C patients had elevated serum HA values (p=ns). Serum HA levels correlated with direct bilirubin level. The control group had lower levels (4.8 [2.3] ng/mL; p< 0.05), which were in the normal range. CONCLUSION: Serum HA level may be useful as a diagnostic tool in children with cirrhosis. PMID- 15333967 TI - Efficacy of omega-3 fatty acids, atorvastatin and orlistat in non-alcoholic fatty liver disease with dyslipidemia. AB - AIM: To evaluate the efficacy and safety of three hypolipidemic agents in patients with non-alcoholic fatty liver disease associated with hyperlipidemia. METHODS: Patients with dyslipidemia (Fredrickson type IIb), asymptomatic persistent transaminasemia lasting 24 weeks, and evidence of hepatic fat infiltration on ultrasonography and liver biopsy were studied. Those with predominant hypertriglyceridemia received omega-3 fatty acids (5 mL thrice daily) (Group A), those with predominant hypercholesterolemia received atorvastatin 20 mg/daily (Group B), and overweight patients received orlistat 120 mg thrice daily before meals (Group C). After 24 weeks of treatment, serum transaminase and lipid levels and liver ultrasonography were repeated. RESULTS: Serum transaminase levels decreased significantly (p< 0.001) in all groups but the decrease was more marked in Group C (AST 75 [16] to 31 [7] IU/L; ALT 120 [38] to 41 [10] IU/L) than in Group A (AST 70 [14] to 41 [6]; ALT 110 [20] to 68 [12]) or Group B (AST 68 [13] to 46 [9]; ALT 115 [22] to 76.6 [13]). After treatment, ultrasonography showed resolution of fatty liver in 35% of patients in Group A, 61% in Group B, and in 86% in Group C (p< 0.001, Group C vs. A). CONCLUSIONS: A decline in transaminase levels and normalization of ultrasonographic evidence of fatty liver were observed on treatment with omega-3 fatty acids in patients with hypertriglyceridemia, with atorvastatin in those with hypercholesterolemia, and orlistat in overweight patients with hyperlipidemia. PMID- 15333968 TI - TTV infection in children with and without liver disease. AB - BACKGROUND: TTV DNA has been reported in patients with a broad spectrum of hepatic disorders as well as in healthy people. AIM: To clarify the role of TTV in children with liver disease and in healthy children. METHODS: Degenerate primers designed to amplify a target sequence from the ORF 1 region of TTV genome were used for nested PCR, to detect TTV DNA in sera. RESULTS: TTV was detected in 3 of 18 children with chronic hepatitis B (16.7%), 2 of 17 hepatitis B carriers (11.8%), 2 of 17 children with cryptogenic chronic liver disease (11.8%), and 1 of 40 (2.5%) children without liver disease. The infection rate was similar among the various study groups and in the various age groups. There was no difference between TTV positive and negative children in respect to gender, history of surgery, parenteral treatment, transfusion of blood and blood products, presence of hepatomegaly, splenomegaly, jaundice, and transaminase values. CONCLUSION: TTV does not seem to have an etiologic role in cryptogenic liver disease in children and does not seem to influence the clinical course of liver disease. PMID- 15333969 TI - Relationship between endoscopic nodular gastritis and Helicobacter pylori infection in children. AB - INTRODUCTION: The specificity of relationship of endoscopic evidence of nodular gastritis with Helicobacter pylori infection is unclear. AIM: To assess the relationship of endoscopic nodular gastritis and H. pylori infection among children. METHODS: 124 children (median age 8.2 years, range 1-15) undergoing upper GI endoscopy for abdominal pain underwent urease test and histological examination of gastric mucosa to determine the presence and density of H. pylori infection, and presence and severity of gastritis. RESULTS: H. pylori infection was detected in 57 (46%) children. Endoscopic nodular gastritis was present in 46 of these 57 patients (81%) and in 24 of 67 (36%) H. pylori-negative patients (36%). The frequency of endoscopic nodular gastritis was related to increasing age (p< 0.0001), presence of H. pylori, grade of histologic gastritis, and H. pylori density (p< 0.0001). CONCLUSION: Endoscopic finding of nodular gastritis is associated with presence of H. pylori infection and active chronic gastritis in children. PMID- 15333970 TI - Natural course of asymptomatic pancreatic pseudocyst: a prospective study. AB - AIM: To study the natural course of asymptomatic pseudocysts of the pancreas. METHODS: Thirty patients (age range 18-68 years, mean 44; 24 men) with asymptomatic pseudocysts of the pancreas were enrolled between December 2001 and December 2003, and were followed up every month. Those who developed symptoms due to pseudocyst (increasing pain or features of obstruction such as vomiting or jaundice) were subjected to an endoscopic or surgical drainage procedure. End point of the study was either spontaneous resolution of pseudocyst or drainage procedure. RESULTS: Eighteen (60%) of 30 patients had resolution of the pseudocyst over an average duration of 5 months. Maximum diameter of less than 7.5 cm and cyst volume less than 250 mL were significantly more frequent in patients with resolution of pseudocyst than in those without (14/18 vs 2/12 [p=0.001] and 15/18 vs 2/12 [p=0.0003], respectively). Presence of internal debris was associated with non-resolution (9/12 vs 2/18; p=0.001). CONCLUSION: Pseudocysts with less than 7.5 cm diameter, volume less than 250 mL, and absence of internal debris were associated with spontaneous resolution within an average duration of 5 months. PMID- 15333971 TI - Repair of duodenal fistula with rectus abdominis musculo-peritoneal (RAMP) flap. AB - INTRODUCTION: Large duodenal defects/fistulae are difficult to repair, due to complex duodenal anatomy. Musculo-epithelial flaps are conventionally used for reconstruction of large soft tissue defects. We report the clinical use of rectus abdominis musculo-peritoneal (RAMP) flap for repair of duodenal fistulae. METHODS: Eight patients with duodenal fistulae underwent repair using right RAMP flap, based on the position of the superior epigastric artery. Feeding jejunostomy was done routinely to start early enteral feeding. RESULTS: Duodenal fistulae healed within 3-5 days in all the cases except one, in whom bile continued to leak for 30 days. One patient died within 12 hours of second surgery. Superficial wound infection was a common (n=3) cause of morbidity. CONCLUSION: RAMP flap for closure of duodenal defect is a simple, technically easy and dependable procedure, which can be performed quickly in critically ill patients. It can be used for repair of large duodenal defects with friable edges when omentum is not available or when other conventional methods are impractical. PMID- 15333972 TI - Hepatobiliary dysfunction as initial manifestation of disseminated cryptococcosis. AB - Disseminated cryptococcosis presenting as biliary obstruction is rare. We report a 35-year-old HIV-negative man who presented with clinical features suggestive of obstructive jaundice, and radiological features suggestive of Klatskin's tumor, but who ultimately was found to have cryptococcal involvement of the liver and biliary tract as part of disseminated cryptococcosis. The patient responded to antifungal therapy. PMID- 15333973 TI - Gastric zygomycosis--an unusual cause of massive upper gastrointestinal bleed. AB - We report a 62-year-old man with cardiac failure and acute renal failure, who had massive hematemesis. Upper GI endoscopy showed a large gastric lesser curvature ulcer. Billroth II gastrectomy specimen showed fungal invasion. He received amphotericin B postoperatively, and recovered uneventfully. PMID- 15333974 TI - Pulmonary aspergillosis in a child with hepatic failure. AB - Invasive aspergillosis is described more frequently as a complication of neoplastic disease and in immunocompromised patients. Hepatic failure is not a generally recognized risk for pulmonary aspergillosis. We report a 3-year-old boy who presented with hepatic failure and pneumonia and whose autopsy revealed liver cirrhosis and pulmonary aspergillosis. PMID- 15333975 TI - Cerebral venous sinus thrombosis as presenting feature of Crohn's disease. AB - Hepatobiliary and vascular manifestations are rare form of extraintestinal manifestations in Crohn's disease. We report a 20-year-old man in whom cerebral venous sinus thrombosis was the presenting symptom and preceded the diagnosis of Crohn's disease. PMID- 15333976 TI - Enterobiasis mimicking Crohn's disease. AB - We report a 20-year-old man who presented with abdominal discomfort for 2 months. Colonoscopy showed skip areas with ulceration, resembling Crohn's disease. Biopsies showed chronic inflammation and a non-necrotizing granuloma. An adult pinworm was found in the lumen from an uninvolved segment. The patient responded to mebendazole. PMID- 15333977 TI - Acute appendicitis presenting as acute hemiscrotum in a boy. AB - A 6-year-old boy presented with diffuse abdominal pain, nausea and vomiting followed by features of acute scrotum. Laboratory and radiological evaluation suggested presence of infective pathology in the inguinoscrotal region. Surgical exploration revealed patent right processus vaginalis with purulent collection resulting from the presence of perforated tip of appendix in the hernial sac. Appendectomy with drainage of scrotal collection and ligation of hernial sac resulted in satisfactory recovery. PMID- 15333978 TI - Post-traumatic retroperitoneal colonic injury presenting as gluteal abscess. AB - We report an 8-year-old boy who sustained blunt retroperitoneal right colonic injury in a vehicular accident and presented with gluteal abscess. Surgical exploration revealed perforated posterior wall of ascending colon with surrounding retroperitoneal abscess communicating with the gluteal region. Right hemicolectomy with drainage of retroperitoneal and gluteal collections resulted in satisfactory recovery. PMID- 15333979 TI - Perforation in a jejunal duplication lined by ectopic gastric mucosa. AB - We report a 3-year-old child with jejunal duplication with localized perforation and bleeding in the adjacent normal intestine. The entire duplicated jejunum was lined by gastric mucosa. This was successfully treated by mucosal excision of the duplicated intestine and resection of a short segment of normal jejunum. PMID- 15333980 TI - Good's syndrome: an unusual cause of chronic diarrhea. AB - A 50-year-old man presented with recurrent episodes of pulmonary infections over a period of 5 years, chronic small bowel diarrhea and weight loss of 6 months' duration. On evaluation he was found to have a thymoma, intestinal infection with giardia, oral candidiasis, and low immunoglobin levels. He was diagnosed to have Good's syndrome. The patient refused further management. PMID- 15333981 TI - Triple therapy using two dosages of metronidazole along with amoxicillin and omeprazole to eradicate Helicobacter pylori infection: a randomized, open study. PMID- 15333982 TI - Determining the point of indifference--where costs of selective and universal immunization against hepatitis B are identical, in a cost-minimization exercise. PMID- 15333983 TI - Umbilical metastasis with squamous cell carcinoma of esophagus. PMID- 15333984 TI - Colonoscopic management of roundworm in cecum. PMID- 15333985 TI - Intraluminal portal vein sign in Caroli's syndrome. PMID- 15333987 TI - [Memantine: a therapeutic drug for Alzheimer's disease and the comparison with MK 801]. AB - Memantine is agreed officially as a therapeutic drug for moderate-to-severe Alzheimer's disease (AD) in EU and USA. Memantine is a similar uncompetitive NMDA receptor antagonist to MK-801 and phencyclidine (PCP), and it prevents nerve cell death induced by the ischemia which induces as excessive release of glutamate. These medicines act on an ion channel binding site similar to the magnesium ion binding site. However, MK-801 and PCP cause schizophrenic symptoms, so they are not being used as a therapeutic drug for AD. Memantine does not have those toxicities and does not stimulate acetylcholine release in the cerebral cortex. Although the mechanism of the difference from memantine and MK-801 has not been made clear yet, it seems that memantine is combined and released with the ion channel depending on electric potential in the same way as the magnesium ion. Basic and clinical research will clarify the control mechanism of memantine. PMID- 15333986 TI - [Possible mechanisms of A beta(1-40)- or A beta(1-42)-induced cell death and their rescue factors]. AB - Amyloidbetapeptide (A beta) is implicated in neuronal cell death in Alzheimer's disease, but the molecular mechanisms are still unclear. We analyzed its mechanism and found several potential rescue factors against A beta-mediated apoptosis. A beta(1-40) stimulated phosphorylation of tau and JNK and induced cell death in SH-SY5Y cells. The cell death was inhibited by insulin-like growth factor-1, suggesting that the JNK pathway may be involved in A beta(1-40)-induced cytotoxicity. Using the human fetus brain cDNA library-targeted differential display technique, a new gene BF5-1 (32aa) was found as a rescue factor against A beta(1-40). BF5-1 has partially the same amino acid sequences as those of the C terminus of cytochrome c oxidase subunit VIIb (COX-VIIb). COX-VIIb mRNA is increased in AD brains and its overexpression in cells enhanced A beta(1-40) toxicity. These data suggest that BF5-1 may act as a dominant negative mutant of COX-VIIb. A beta(1-42) also induced cell death in rat neuroblastoma B104 cells, which was abolished by addition of IL-11. By cDNA subtraction analysis in the cell death, the enhanced expression of L-phosphoserine phosphatase was found, but this was also abolished by IL-11. The glutamate neurotoxicity was stimulated in the presence of D-serine, suggesting that NMDA receptors may be involved in A beta(1-42)-induced cytotoxicity. A beta(1-42) also induced increase of a new gene p18A beta rP (p18-amyloid-beta-responsive protein; 166 aa) mRNA expression; overexpression of this gene in PC12 cells induced cell death. By the application of a death trap method, a new gene, p60TRP (p60-Transcription-Regulating-Protein; rat:539 aa, human:547aa), was found as a potential rescue factor against the cell death by p18A beta rP. Thus, our cell death systems and/or new rescue proteins may provide suitable tools for the establishment of drug screening systems leading to the identification of new low-molecular candidates applicable for the treatment of AD. PMID- 15333988 TI - [Pre-clinical evaluation of effects of acetylcholinesterase inhibition on the cerebral cholinergic neuronal system and cognitive function: PET study in conscious monkeys]. AB - The present review described the effects of acetylcholinesterase (AChE) inhibition on the cerebral cholinergic neuronal system in the conscious monkey brains with PET. Somatosensory stimulation induced a regional cerebral blood flow (rCBF) response, revealed with [(15)O]H(2)O, in the contralateral somatosensory cortex. Scopolamine resulted in an abolished rCBF response to stimulation, and this abolished rCBF response was recovered by physostigmine, donepezil, and tacrine. Donepezil suppressed AChE activity, analyzed by [(11)C]MP4A, in all cortical regions in a dose-dependent manner. AChE inhibition by donepezil resulted in a dose-dependent increase in acetylcholine levels in the prefrontal cortex as measured by microdialysis. Binding of [(11)C](+)3-PPB to cortical muscarinic receptors was reduced by donepezil, probably in a competitive inhibition manner. Aged monkeys showed less reduction of [(11)C](+)3-PPB binding than young animals. As evaluated by an oculomotor delayed response task, aged monkeys showed impaired working memory performance compared to young monkeys, and the impaired performance was partly improved by the administration of donepezil, due to the facilitation of the cholinergic neuronal system by AChE inhibition by donepezil. These results demonstrated that PET imaging with specifically labeled compounds in combination with microdialysis and a behavioral cognition task could be a useful tool for pre-clinical evaluation of novel drugs. PMID- 15333989 TI - [Scope and limitations of acetylcholinesterase inhibitors]. AB - Enhancement of the activity of cholinergic neurons has been regarded as one of the most promising methods for treating Alzheimer's disease (AD). Donepezil is a representative acetylcholinesterase inhibitor (AChEI) and is a great success among the AChEI drugs. AChEIs are being studied for other mechanisms of action, neuroprotective action, and nicotinic receptor enhancement. AD is a type of neurodegenerative disease and AChEIs have been found to be an effective anti-AD medication. AChEI can alleviate the symptoms and delay the progression of AD, but it cannot cure the disease. However, AChEIs are now the subject of a wide range of clinical studies for other diseases, for example, other types of dementia (such as Lewy body disease, cerebral vascular dementia, and Parkinson's disease dementia), and migraine. These drugs are also being studied as a combination therapy, for example, with an antioxidant, SERM, and NMDA antagonist. PMID- 15333990 TI - [Dexmedetomidine hydrochloride (Precedex), a new sedative in intensive care, its pharmacological characteristics and clinical study result]. AB - Dexmedetomidine hydrochloride (Precedex) is a potent and highly selective central alpha(2)-adrenoreceptor agonist. In a test of affinity to receptors in rat brain cortex, this drug showed a high affinity and selectivity to the alpha(2) adrenoreceptor. This drug induced a dose-dependent decrease in activity, loss of righting reflex, an increase in sedation score, and a dose-dependent prolongation of the latent time of escape from pain in various animal models. The site of action for the sedative action of this drug is considered to be the locus coeruleus. With administration of this drug into the locus coeruleus, loss of righting reflex was observed in almost all animals. Test results using mice with variant alpha(2A)-receptor suggested that the sedative action of this drug is expressed through the alpha(2A)-receptor subtype. This drug is promptly eliminated from blood. In the phase II/III multi-center placebo-controlled double blind bridging study conducted in Japan, efficacy and safety were examined in patients brought to the ICU. As regards sedative action, the ratio of patients who did not require additional propofol medication (>50 mg) while intubated was taken as the effective rate. The drug group showed a significantly high effective rate (90.9%; placebo group, 44.6%). Similarly, with analgesic action, the ratio of patients who did not require additional morphine medication during intubation was taken as the effective rate. The effective rate for the drug group was significantly high (87.3%; placebo group, 75.0%). The main adverse events observed in the drug group were hypertension and hypotension. PMID- 15333991 TI - FMRFamide-expressing efferent neurons in eighth abdominal ganglion innervate hindgut in the silkworm, Bombyx mori. AB - The tetrapeptide FMRFamide is known to affect both neural function and gut contraction in a wide variety of invertebrates and vertebrates, including insect species. This study aimed to find a pattern of innervation of specific FMRFamide labeled neurons from the abdominal ganglia to the hindgut of the silkworm Bombyx mori using the immunocytochemical method. In the 1st to the 7th abdominal ganglia, labeled efferent neurons that would innervate the hindgut could not be found. However, in the 8th abdominal ganglion, three pairs of labeled specific efferent neurons projected axons into the central neuropil to eventually innervate the hindgut. Both axons of two pairs of labeled cell bodies in the lateral rind and axons of one pair of labeled cell bodies in the posterior rind extended to the central neuropil and formed contralateral tracts of a labeled neural tract with a semi-circular shape. These labeled axons ran out to one pair of bilateral cercal nerves that extended out from the posterior end of the 8th abdominal ganglion and finally to the innervated hindgut. These results provide valuable information for detecting the novel function of FMRFamide-related peptides in metamorphic insect species. PMID- 15333992 TI - Desiccation tolerance of the tardigrade Milnesium tardigradum collected in Sapporo, Japan, and Bogor, Indonesia. AB - A tardigrade Milnesium tardigradum showed anhydrobiotic capacity, in which the desiccation tolerance, given by the mean survival rate under desiccation at different relative humidity levels, was significantly higher in the Sapporo (Japan) population than that in the Bogor (Indonesia) population. Accordingly, the surviving tardigrades took a significantly longer time for revival in Bogor than those in Sapporo. The higher tolerance of the Sapporo population is thought to be related to the low relative humidity and low temperature such that the animals experience 41% RH in May and often -10 degrees C or lower in winter. PMID- 15333993 TI - Sexual dimorphism of acoustic signals in the oriental white stork: non-invasive identification of sex in birds. AB - Identification of the sex of birds is important for captive breeding of endangered species. In the oriental white stork (Ciconia boyciana), an endangered species, both sexes produce an acoustic signal called "clatter" by rattling their mandibles together to generate sounds. We examined the structure of male and female clatter to determine whether clatter is sexually dimorphic. The acoustic structure of the clatter of the two sexes proved to be dimorphic with respect to the fundamental frequency; female clatter had higher fundamental frequencies. The fundamental frequency correlated significantly and positively with bill length, suggesting that bill morphology contributes to the sexual dimorphism of clatter. Sexing can be done by acoustic signals without capturing birds, and thus is useful as a non-invasive sexing method for ecological and conservation studies of birds. PMID- 15333994 TI - Defense function of pigment granules in the ciliate Blepharisma japonicum against two predatory protists, Amoeba proteus (Rhizopodea) and Climacostomum virens (Ciliata). AB - The defense function of pigment granules in the red ciliate Blepharisma japonicum against two predatory protists, Amoeba proteus and Climacostomum virens, was investigated by (1) comparing normally-pigmented and albino mutant cells of B. japonicum as the prey of these predators and (2) comparing resistance of the predators to blepharismin, the toxic pigment contained in the pigment granules of B. japonicum. Normally pigmented cells which contained more blepharismin than albino cells were less vulnerable to A. proteus than albino cells, but not to C. virens. C. virens was more resistant than A. proteus to the lethal effect of blepharismin. The results indicate that pigment granules of B. japonicum function as defense organelles against A. proteus but not against C. virens and suggest that successful defense against a predator depends on the susceptibility of the predator to blepharismin. PMID- 15333995 TI - Novel development rescuing factors (DRFs) secreted by the developing Dictyostelium cells, that are involved in the restoration of a mutant lacking MAP kinase ERK2. AB - We found novel development rescuing factors (DRFs) secreted from developing Dictyostelium cells, by using a mutant (erkB-) which is missing MAP-kinase ERK2 as a test strain for bioassay. The mutant erkB- fails to undergo multicellular morphogenesis due to impaired cAMP signaling. However, such developmental defect can be restored by the presence of low-molecular weight DRFs that are secreted from developing wild-type cells. We previously showed that DIF-1 (Differentiation Inducing Factor 1 for stalk cells) possesses this activity, indicating a newly discovered role of DIF-1. Surprisingly, however, the mutant dmtA-, which is incapable of DIF-1 synthesis still exerts a strong inducing activity of the multicellular morphogenesis of erkB-. After analysis of HPLC fractions of conditioned media prepared from both wild type Ax2 and dmtA- strains revealed that both strains secrete at least two novel DRF activities with DIF-like mobility. However, these activities were not derived from other DIFs such as DIF 2 and DIF-3. Identification of these DRFs found in this study would provide insight into the mechanism by which the development of the erkB- mutant is restored and how these factors act in the normal development of Dictyostelium. PMID- 15333996 TI - Photoperiod- and temperature-dependent regulation of pupal beige/black polymorphism in the small copper butterfly, Lycaena phlaeas daimio Seitz. AB - The small copper butterfly, Lycaena phlaeas daimio, has pupal beige/black polymorphism, the development of which is found to be controlled in an apparent association with the development of adult seasonal polymorphism (spring and summer morphs) by photoperiod and temperature in the larval stages. That is, the pupae of beige and black types developed under long-day and short-day conditions tend to develop into brown-winged and red-winged adults, respectively. In addition, a large proportion of long-day pharate pupae chilled at 4 degrees C for 5 days were observed to develop into pupae whose head-thoracic complexes and abdomens were judged to be of the black and intermediate types, respectively. They developed into adults with redder wings as compared to those obtained from unchilled pupae. The results indicate that the physiological mechanism underlying the photoperiodic control of the development of adult seasonal polymorphism may also play a significant role in the determination of pupal beige/black polymorphism in L. phlaeas daimio. Furthermore, cuticle melanization was found to be induced in the head-thoracic complexes of pupae by chilling of the pharate pupae. Melanization of pupal cuticle seems to occur in a close association with the development of reddish-winged adults. PMID- 15333997 TI - Comparative histological study of teleost livers in relation to phylogeny. AB - This report presents a detailed description of hepatic architecture in 200 teleost livers by light microscopy and extensively discusses the phylogenetic viewpoint. The 200 teleost livers showed a great variety of histological images, but not the same image, as in mammalian livers. The hepatocyte-sinusoidal structures of the fish livers were classified into three different types: (a) cord-like form, (b) tubular form, (c) solid form. Biliary tract structures were classified into four types: (a) isolated type, (b) biliary-arteriolar tract (BAT) type, (c) biliary-venous tract (BVT) type, and (d) portal tract type. As phylogenic advancement is graded from low to high, the parenchymal arrangement progressed from solid or tubular form to cord-like form, but the biliary tract structures were not involved. We demonstrate that this study is the first to investigate teleost livers phylogenically, and their architectural differences are shown in the route of hepatic ontogenesis. In hepatic ontogenesis, the formation of the parenchymal arrangement is acquired phylogenically, but the biliary pathway may be adapted in the ecological and behavioral patterns. PMID- 15333998 TI - Possible involvement of a sperm-associated body in the process of fertilization in quail. AB - The present paper describes a novel structure, termed the sperm-associated body, which is found both in the lumen at the oviductal infundibulum and in the vitelline membrane of the ovum in the quail Coturnix japonica. The fully developed sperm-associated body, which is about 100 microm long, consisted of two parts; a core of concentric-circular appearance and a cortex of needle-like projections. The outer surface of the body was coated with CaCO3. The body was always accompanied by spermatozoa. About 70 sperm-associated bodies were observed in a single ovum. Electron-microscopically, small numbers of holes were detected in the vitelline membranes of a fertile ovum, and the sperm-associated bodies were always present in these holes. Frequently observed in the vitelline membranes was a disk speculated to be a portion of the inner layer of the membrane partially affected by spermatozoa. However, neither sperm-associated bodies nor spermatozoa were observed there. It was suggested that the sperm associated bodies assist fertile spermatozoa in binding the inner layer of the vitelline membrane and penetrating it. PMID- 15333999 TI - Female mate preference in goby Eviota prasina: do secondary sexual traits influence female choice? AB - Males of the small gobiid fish Eviota prasina have longer dorsal fins than females and use their dorsal fins for courtship displays and male-male competition. We examined the influence of male dorsal fin length as well as the frequency of courtship displays on female mate preference using sequential choice experiments. Females responded more frequently to courting males with longer dorsal fins than males with shorter dorsal fins. This indicates female mate preference on the basis of male dorsal fin length in this goby. In addition, the logistic regression analysis showed that male courtship frequency positively influenced female mate preference. Since males incur costs to produce and maintain longer dorsal fins and to perform frequent courtship displays, these secondary sexual traits may indicate the quality of physical characteristics. In E. prasina, only males conducted parental care for eggs in their nests. Therefore, females may be able to ensure greater survival rates as well as higher genetic quality of their offspring by choosing males with better qualities based on these male traits. PMID- 15334000 TI - Periclimenes speciosus, a new species of anthozoan associated shrimp (Crustacea: Decapoda: Palaemonidae) from southern Japan. AB - A new species of palaemonid shrimp, Periclimenes speciosus sp. nov., is described and illustrated on the basis of 20 specimens collected from warm-temperate and subtropical waters of southern Japan. Periclimenes speciosus belongs to the "P. aesopius species group", and is associated with sea anemones and a scleractinian coral as well as behaving as a fish cleaner. Morphologically, the new species appears closest to P. holthuisi Bruce, 1969, but can be distinguished from P. holthuisi by the form and armature of the cutting edges of dactylus and fixed finger of the second pereiopod. The coloration in life of both species clearly discriminates one species from the other. The taxonomic status of some specimens previously reported as P. holthuisi is briefly discussed. PMID- 15334001 TI - Cell number and cellular composition in infusoriform larvae of dicyemid mesozoans (Phylum Dicyemida). AB - Cell numbers and cellular composition were examined in infusoriform larvae of 44 species of dicyemid mesozoans belonging to 6 genera; Conocyema, Dicyema, Dicyemennea, Dicyemodeca, Microcyema, and Pseudicyema. In addition, literature on infusoriform larvae of another 20 species was reviewed. Infusoriform larvae consist of a constant cell number which is species-specific. Small interspecific variations are found in total cell numbers, 35, 37, 39, 41 and 42. The most frequent cell number encountered in infusoriform larvae studied is either 37 or 39. Infusoriform larvae with 35 cells are found in three genera. Infusoriform larvae with 37 cells are found in four genera. Infusoriform larvae with 39 cells are found in four genera. Most differences in total cell numbers are due to the absence or presence of particular ventral cells. In all infusoriform larvae, the lateral, dorsal and caudal areas are cell constant, whereas in the apical and ventral areas a distinct and variable configuration of cells are present. In cellular composition, a total of 29 cells (15 cell types) were recognized in all infusoriform larvae examined. Additional cell types are characteristic of a relatively few species. Even in infusoriform larvae with the same total cell numbers, cellular composition varies by species. Thus, there are 7 variations of cellular composition in infusoriform larvae with 37 cells. Differences in larval cell numbers and types do not warrant traditional generic separation of dicyemids. PMID- 15334002 TI - Study of color variation in the solitary ascidian Halocynthia roretzi, collected in the Inland Sea of Japan. AB - In the vicinity of Yashiro Island in the Inland Sea of Japan, the solitary ascidian (tunicate) Halocynthia roretzi with tunics of various colors were collected. Samples of these animals were sorted into three groups on the basis of visual observation of tunic color. The red group includes animals with dark-red, light-red, or orange tunics. The pink group includes animals with tunic colors ranging between red and white. The white group includes only animals with completely white tunics. Animals in the white group lacked color internally, with the exception of the hepatopancreas and the gonads in breeding season; the epidermis and gill basket were white. In contrast, animals of both the red group and the pink group were colored internally, with red-orange epidermis and yellow gill basket. Alloreactivity was tested by mixed-hemocyte incubation between different animals belonging to the same color group and between animals belonging to different color groups. Alloreactivity between animals of the white group was 56.3%, between animals of the pink group was 60.0%, and between animals of the red group was 69.3%. The relatively high frequency of compatible combinations among the white animals is discussed. PMID- 15334003 TI - Poorly differentiated small cell carcinoma of the pancreas. A case report and review of the literature. AB - Small cell carcinoma (SCC) of the pancreas is a rare malignancy with an extremely poor prognosis. We present the case of a 74-year-old man with a 2-month history of upper abdominal discomfort who was diagnosed with SCC of the pancreas tail, involvement of peripancreatic and mesenteric lymph nodes and multiple liver metastases (extended disease). A CT scan and a positive somatostatin receptor scintigraphy showed no evidence of a primary lung tumour. The diagnosis of a SCC was confirmed by biopsy. Local tumour control could be achieved by gemcitabine once a week and a long-acting somatostatin analogue once a month, but liver metastasis showed progress. Thus, 5-fluorouracil on a weekly basis was started. The patient died 8 months after diagnosis and had not been hospitalised in the meantime. PMID- 15334004 TI - Mechanism of acute pancreatitis exacerbation by enteral bile-pancreatic juice exclusion. AB - Using an original model, the donor rat model, we previously showed that bile pancreatic juice exclusion from gut exacerbates ligation-induced acute pancreatitis. Here, we examine the mechanism by which bile-pancreatic juice exclusion from gut exacerbates acute pancreatitis. In the first part of the study we test the hypothesis that Na taurocholate and trypsin are components of bile pancreatic juice that exacerbate acute pancreatitis when excluded. Our experiments show that combined replacement of Na taurocholate and trypsin ameliorates acute pancreatitis. In the second part of the study we test the hypothesis that bile-pancreatic juice exclusion from gut exacerbates acute pancreatitis via combined CCK-A and cholinergic receptor pathways. Our experiments show that combined CCK-A and cholinergic receptor blockade significantly ameliorates acute pancreatitis while blockade of either receptor alone does not. We conclude that bile-pancreatic juice exclusion-induced exacerbation of ligation-induced acute pancreatitis involves a neurohormonal duodenal response to exclusion of trypsin and Na taurocholate resulting in acinar cell hyperstimulation via combined CCK-A and cholinergic receptor-mediated pathways. PMID- 15334005 TI - Risk profiles for ageing macular disease. PMID- 15334006 TI - Pathogenesis of diabetic retinopathy and strategy to develop new therapeutic modalities. PMID- 15334007 TI - Therapeutic and cosmetic indications of lotrafilcon a silicone hydrogel extended wear lenses. PMID- 15334008 TI - Medical treatment of retinopathy of type-2 diabetes. AB - The medical treatment of retinopathy in type-2 diabetes should be considered as a major component in the overall management of diabetic retinal disease. It is clear that specific and timely interventions, such as glycemic and blood pressure control, are the basis for good management of diabetic retinopathy. The American Diabetes Association has developed specific recommendations concerning diabetic retinopathy for the primary care physician and diabetologist. The ophthalmologist must be aware of these recommendations and establish efficient communication channels with the colleagues who follow their patients and the progression of diabetes closely. The challenge for the ophthalmologist is to make sure that, when signs of retinopathy are detected, information regarding the status of the retina, prognostic factors and the rate of progression must be given to the primary care physician and diabetologist. Under these circumstances, excellent glycemic control, aggressive management of blood pressure and normalization of lipids are all needed, and the goals to be achieved must be shared between the ophthalmologist and the primary care physician or diabetologist. Appropriate medical management of diabetic retinopathy is fundamental to reduce the risk of blindness. This goal can only be achieved if the ophthalmologist is fully aware of the role of medical management and establishes an efficient flow of communication with the primary care physician or diabetologist, particularly for the diabetic patients whose eyes show signs of risk for rapid progression of their retinopathy. PMID- 15334009 TI - Double visualization using triamcinolone acetonide and trypan blue during stage 3 macular hole surgery. AB - PURPOSE: To study the usefulness of intravitreal injection of triamcinolone acetonide and trypan blue for facilitating visualization and dissection of the posterior vitreous cortex and internal limiting membrane (ILM) during vitrectomy in idiopathic stage 3 macular holes. METHODS: Pars plana vitrectomy was performed in 10 eyes of 10 patients with idiopathic stage 3 macular holes. After core vitrectomy had been performed, triamcinolone acetonide was injected over the posterior pole. After separation of the visualized posterior vitreous cortex, trypan blue was injected over the macular area. Excised specimens were examined by electron microscopy. RESULTS: Upon injection of triamcinolone acetonide, the posterior vitreous cortex and residual vitreous cortex could be visualized in all patients. The posterior vitreous cortex and residual vitreous cortex were completely removed. The ILM of the retina was stained faint blue and was successfully removed in all patients. Electron microscopy revealed that the triamcinolone-acetonide-visualized layer and the trypan-blue-stained layer had different histological features. No complications related to the use of triamcinolone acetonide and trypan blue were encountered. CONCLUSION: Double visualization of the posterior vitreous cortex and ILM using triamcinolone acetonide and trypan blue during vitrectomy may facilitate separation of the posterior vitreous cortex from the retina and removal of the ILM around the macular hole in patients with idiopathic stage 3 macular holes. PMID- 15334010 TI - Endoscopic transnasal dacryocystorhinostomy and bicanalicular silicone tube intubation. AB - PURPOSE: To evaluate the results of endoscopic transnasal dacryocystorhinostomy (ETDCR) combined with bicanalicular silicone tube intubation (BSTI) performed in 49 patients with nasolacrimal canal obstructions secondary to chronic dacryocystitis. MATERIAL AND METHODS: ETDCR combined with BSTI was applied as a primary procedure in 47 cases, and as a secondary procedure in 2 cases. All patients underwent dacryocystography, and 41 patients underwent radionuclide dacryoscintigraphy before the surgery. Silicone tubes were left in place for about 6.2 months. The patency of the lacrimal passages was controlled with a clinical examination, nasolacrimal canal lavage, and dacryoscintigraphy. RESULTS: Patients were followed up for a mean period of 25.1 months after the removal of the tubes. The epiphora symptom disappeared and a successful drainage could be maintained in 43 out of 49 cases (87.7%). The obstruction rate found in the dacryoscintigraphy performed 15 days after extubation was 22.9%, while it was found to be 14.2% in the nasolacrimal canal lavage. CONCLUSION: ETDCR combined with BSTI proved to be an effective surgical method in chronic dacryocystitis. The sensitivity and specificity of the postoperative dacryoscintigraphy in determining the surgical success were lower than those of the nasolacrimal canal lavage. PMID- 15334011 TI - Prostaglandin analogs and blood-aqueous barrier integrity: a flare cell meter study. AB - PURPOSE: To study, with an objective method, inflammation of the anterior segment of the glaucomatous eye after treatment with latanoprost, travoprost and bimatoprost. MATERIALS AND METHODS: Sixty patients with chronic open-angle glaucoma aged between 38 and 76 years (mean 64.0 +/- 12.2) were randomly assigned to latanoprost 0.005, travoprost 0.004 and bimatoprost 0.03%. The study period lasted 6 months. Intraocular pressure (IOP) was measured every 2 weeks. We studied the intraocular inflammation before and after 3 and 6 months of therapy with an instrument composed of a He-Ne laser beam system, a photomultiplier mounted on a slitlamp microscope and a computer. This flare meter allows objective determination of the flare and the number of cells in the aqueous of the anterior chamber. RESULTS: At the baseline, IOP was 26.4 +/- 3.6 mm Hg. After 3 months of treatment, mean IOP in the latanoprost group was 17.9 +/- 0.3 mm Hg (p < 0.001) with a mean cellularity of 12.638 +/- 3.284 photons/ms (p < 0.001). The travoprost group had an IOP of 17.2 +/- 0.3 mm Hg (p < 0.001) with a cellularity of 9.719 +/- 1.927 photons/ms (0.001). Finally, IOP in the bimatoprost group was 17.6 +/- 0.5 mm Hg (p < 0.001) with a cellularity of 6.138 +/- 1.475 photons/ms (p < 0.032). After 6 months of treatment, IOP in the latanoprost group was 18.1 +/- 0.3 (p < 0.001), in the travoprost group 17.3 +/- 0.3 (p < 0.001) and in the bimatoprost group 17.7 +/- 0.5 mm Hg (p < 0.001), whereas cellularity was 11.838 +/- 3.218 (p < 0.001), 8.950 +/- 3.692 (p < 0.001) and 7.617 +/- 2.603 photons/ms (p < 0.001), respectively. After 3 months, the travoprost (p < 0.013) and the bimatoprost groups (p < 0.001) had less flare compared with the latanoprost group and this remained so even at 6 months. When we compared the travoprost group with the bimatoprost group, we found significantly less flare at 3 months in the bimatoprost group (p < 0.001) but not at 6 months (p < 0.246). CONCLUSIONS: The flare meter analysis shows that the eyes treated with bimatoprost and travoprost have a less significantly broken blood-aqueous barrier and their anterior chamber is also significantly less inflamed. PMID- 15334012 TI - The efficacy of hourly prophylactic steroids in diffuse lamellar keratitis epidemic. AB - PURPOSE: To investigate the inciting agent, clinical features, and the efficacy of hourly steroids in the prophylaxis in a diffuse lamellar keratitis (DLK) epidemic. METHODS: One hundred and five eyes of 58 patients that had DLK after LASIK were included in the study. To identify the cause of the epidemic, some interventions were made: irrigation solution was changed from BSS to Ringer lactate (week 5), wiping of the interface with a sponge was discontinued (week 6), the air conditioner in the LASIK room was checked (week 6), the routine postoperative topical regimen was changed from fluoromethalone and fluoroquinolones 4 times a day to hourly prednisolone acetate or dexamethasone sodium and fluoroquinolones (week 7), and the trademark of the drape used was changed (week 12). RESULTS: There was no statistical difference in any of the attack rates associated with variables including BSS versus Ringer lactate (9.9 vs. 14%, p = 0.4), and air conditioner check with wiping versus not wiping the interface with the microsurgical sponge (14.0 vs. 18.2%, p = 0.6). There was a significant decrease in the attack rate from 18.2 to 5.3% with use of the hourly prophylactic topical steroids (p = 0.012). After introduction of a new trademark of the drape, the incidence of DLK was further reduced from 5.3 to 0.7% (p = 0.016). CONCLUSION: The cause in a particular DLK epidemic should be identified and eliminated adopting a scientific approach. Hourly steroid use for prophylaxis is recommended until the etiologic agent responsible has been identified. PMID- 15334013 TI - Antifungal activity of intraocularly used liquids against Aspergillus. AB - The antifungal effectiveness of liquids used as intraocular tamponading agents in vitrectomy was tested against a strain of Aspergillus niger. This microorganism is a frequent causative factor of endophthalmitis. The strain belonged to the ATCC collection (A. niger ATCC 16404). The samples tested were: (a) perfluorocarbons: perfluorodecalin and perfluoroctane, (b) silicone oils: Siloil 1,000 and Siloil 5,000, and (c) balanced salt solutions: BSS and BSS Plus. The experiment consisted in inoculating the samples with the microorganism, in preserving the samples in 25 degrees C in the dark, and in taking small amounts of each product for counting surviving microorganisms at t = 0 and then after 1 day, and 1, 2, 3 and 4 weeks. Among the samples tested, perfluorocarbons and silicone oils conform to the standards. No increase, and in some samples decrease, in the number of microorganisms is observed during the second week, while the number drops to zero at the end of the experiment. Therefore, the use of these liquids is safe. PMID- 15334014 TI - The HIV RNA Levels of Plasma and Ocular Fluids in AIDS Patients with Ophthalmic Infections. AB - The aim of the present study was to evaluate the impact of highly active antiretroviral therapy (HAART) on HIV viral load of plasma and intraocular fluids in AIDS patients with ophthalmic opportunistic infections. We further compared the treatment effect of HAART on these patients. From June 1997 to July 2003, we examined and followed up the ophthalmic conditions of 49 patients receiving HAART with ophthalmic diseases during this period. The method of reverse-transcriptase polymerase chain reaction was used to detect and monitor HIV load in plasma and/or aqueous humor of AIDS patients. Before HAART, the HIV levels in the plasma and aqueous humor in 8 AIDS patients with ophthalmic opportunistic infections were significantly higher than those in 6 patients with HIV-related retinopathy (p < 0.05). Compared to the eye findings and clinical improvement, HIV loads of aqueous humor in 10 of 14 AIDS patients (6 with HIV-related retinopathy, 5 with cytomegalovirus retinitis, 2 with toxoplasmic retinitis and 1 with cryptococcal chorioretinitis) declined to undetectable levels (< 400 copies/ml) after 4-8 months of HAART. HIV virus levels in the plasma of AIDS patients were significantly decreased, and the CD4 counts of these patients were significantly increased (Wilcoxon test) after initiation of HAART. PMID- 15334015 TI - Posterior vitreous detachment: clinical correlations. AB - BACKGROUND: To evaluate the frequency of, and the factors which are primarily and secondarily associated with, posterior vitreous detachment (PVD), and to correlate the presence of PVD with various demographic and ophthalmic entities. METHODS: The clinical prospective observational cohort study included 1,481 subjects (740 women, 741 men) with a mean age of 63.45 +/- 14.97 years (mean +/- SD; range, 10.3-94.9 years) and a mean spherical equivalent refractive error of +0.68 +/- 2.13 dpt (range, -14.25 to +13.50 dpt). The presence of PVD was assessed by indirect and direct ophthalmoscopy, fundus biomicroscopy and by using a Hruby lens upon pharmacologically dilated pupil. Main outcome measures were frequency of PVD, association with age, refractive error, cataract surgery, diabetes mellitus, arterial hypertension, history of ocular trauma, vitreous hemorrhage, various retinal disorders, nonarteritic anterior ischemic optic neuropathy (AION) and open-angle glaucoma. RESULTS: The occurrence of PVD was significantly correlated with increasing age (p < 0.001), myopic refractive error (p < 0.001), female gender (p < 0.001), and surgical aphakia (p < 0.001). PVD occurred significantly more often bilaterally than unilaterally (p < 0.001). Patients with unilateral PVD were significantly (p < 0.001) younger than the patients with bilateral PVD. PVD was seen significantly (p = 0.038) less frequently in AION than in the remaining study population. The frequency of PVD was lower in eyes with than without diabetic retinopathy (p = 0.095), optic disc neovascularization (p = 0.07) and retinal neovascularization (p = 0.09). There was no significant association between the presence of PVD and macular hole, macular edema, retinal vascular occlusive disorders, age-related macular degeneration and open-angle glaucoma. CONCLUSIONS: These data of a large prospective study of PVD confirm some of the findings of previous smaller and retrospective studies. They also provide new information which may be helpful for the understanding of the pathophysiology of PVD and the role of PVD in the pathogenesis of various retinal and optic disc lesions. PMID- 15334016 TI - Review of the ischemia hypothesis for ocular hypertension other than congenital glaucoma and closed-angle glaucoma. AB - Experimentally induced ischemia of the endothelial cells surrounding the aqueous drainage sites led to a rise in intraocular pressure (IOP) in monkey and pigeon eyes. Clinical conditions associated with a rise in IOP in human eyes, e.g. peripheral retinal detachments, subluxated lenses, and occlusion of the internal carotid artery, can result in ischemia of the endothelial cells lining Schlemm's canal. These findings led to the hypothesis that ischemia of the endothelial cells of Schlemm's canal induces hypertension in human eyes. Thus, the ischemia hypothesis should be considered in discussions of the etiology of age-related open-angle glaucoma. PMID- 15334017 TI - Age-related modifications of corneal sensitivity. AB - PURPOSE: To evaluate central and peripheral corneal sensitivity (CS) in relation to age. METHODS: Five hundred eyes of 320 healthy subjects (185 males and 135 females) were examined. The age of participants ranged from 20 to 90 years. All subjects were divided into 3 groups according to age. CS was assessed with the Cochet-Bonnet esthesiometer in the central cornea and in 8 peripheral points. The averages were used as the values of central and peripheral CS. RESULTS: In the young population, CS was equal in both examined zones (p > 0.05); although, with respect to the periphery after the fifth decade, it was significantly higher in the centre (p < 0.05). Both central and peripheral sensitivity decreased with age (p < 0.05), and such variations are represented by parabolic curves. No differences were observed between males and females. CONCLUSIONS: The age-related decrement of CS involves at first the corneal periphery and successively spreads toward the central zone. Topographical distribution and age-related modifications should be considered in clinical investigations of CS, especially in patients who underwent any corneal surgical procedure. PMID- 15334018 TI - Ciliary body adenoma after application of mitomycin. AB - PURPOSE: To describe a ciliary body tumor that was detected in a rabbit eye. METHODS: For a specific experiment, the rabbit underwent trabeculectomy with mitomycin. The eye was evaluated by electron microscopy 5 weeks following surgery. RESULTS: A ciliary body tumor was discovered in close relation to pigmented epithelial cells. The tumor cells were large with a small amount of pigment and contained prominent vesicles. CONCLUSIONS: The development of a ciliary body adenoma may be caused by the application of mitomycin. Whether the demonstrated tumor was benign or malignant could not be determined. While it could not be proven that the tumor developed related to the use of mitomycin, this possibility may be important for surgeons using this substance. PMID- 15334019 TI - IUI and evidence-based medicine: an urgent need for translation into our clinical practice. PMID- 15334020 TI - Should we continue performing intrauterine inseminations in the year 2004? AB - This review summarizes the existing evidence regarding intrauterine insemination (IUI) as a treatment for cervical hostility, male and unexplained subfertility. IUI in natural cycles has been proven effective in patients with cervical hostility and moderate male subfertility. IUI in cycles with mild ovarian hyperstimulation (MOH) should be the treatment of choice in couples with mild male subfertilty (average total motile sperm count above 10 million) and unexplained subfertilty. When MOH is applied, gonadotropins have been proven more effective compared with clomiphene citrate. Further large trials comparing clomiphene citrate with gonadotropins are mandatory. Prevention of multiple pregnancies in MOH/IUI programs is of paramount importance. A strategy with a low dose step-up protocol and strict cancellation criteria is proposed. When multiple pregnancies are kept to a minimum, MOH/IUI is more cost-effective compared with in vitro fertilization and embryo transfer. Future research should focus on prediction models to predict the outcome of MOH/IUI treatment for individual couples before starting treatment. PMID- 15334021 TI - The construction of a new evaluative GERD questionnaire - methods and state of the art. AB - Gastroesophageal reflux disease (GERD) is one of the most prevalent diseases worldwide, and it is becoming increasingly important to monitor the effect of various interventions on GERD symptoms. There can be rapid temporal changes in the severity and frequency of patients' symptoms as well as their health status and well-being, all of which could, theoretically, be monitored using diaries or questionnaires. However, current GERD monitoring instruments are not appropriate because they do not assess symptoms daily, they are not sufficiently responsive to short-term changes in health status or they are not adequately validated. To address these problems, the conceptual and psychometric requirements for a GERD symptom assessment questionnaire were identified. A dimension-based scale was designed to reduce the number of symptoms monitored on a daily basis, and the validation process was defined to produce parallel long and short forms of a scale for patients' self-assessment of their GERD symptom response to therapy. These basic principles which underlie the successful development of a new, self assessed symptomatic reflux questionnaire (ReQuest(TM)) are also applicable to the development of validated questionnaires for daily symptom self-assessment in other disease areas. PMID- 15334022 TI - Takotsubo cardiomyopathy: a little-known cardiomyopathy makes its US debut. PMID- 15334023 TI - Family history of coronary artery disease and prognosis after first acute myocardial infarction in a national survey. AB - A positive family history (FH) of coronary artery disease (CAD) is considered an independent risk factor for developing CAD. However, the natural history, coronary angiographic findings and prognosis of patients with a positive FH developing first acute myocardial infarction (AMI) are not well defined. A cohort of 2,690 consecutive patients with first AMI from two prospective nationwide surveys conducted during 1996 and 1998 in all coronary care units operating in Israel was studied. Baseline characteristics, hospital course, management and outcome of 405 patients with first AMI and a positive FH were compared with 2,285 controls without a positive FH. Coronary angiograms of patients with and without a positive FH were reviewed and compared. Patients with a positive FH were younger (53 vs. 64 years), more often male, current smokers and patients with hyperlipidemia, but less often patients with diabetes or hypertension than patients without a positive FH. Patients with a positive FH developed heart failure during hospital stay less frequently. Thrombolytic therapy was similarly administered to both groups. During the hospital stay, coronary angiography, percutaneous coronary intervention or coronary artery bypass grafting were more frequently performed in patients with a positive FH. The coronary anatomy and the extent of the CAD were similar in patients with and without a positive FH. Crude and covariate-adjusted mortality rates were significantly lower in patients with a positive FH than in patients without a positive FH on day 30 (2.2 vs. 9.6%, p < 0.001; odds ratio 0.50, 95% confidence interval 0.22-0.99) and at 1 year (3.5 vs. 14%, p < 0.001; hazard ratio 0.58, 95% confidence interval 0.42-0.80). Patients with a positive FH developed their first AMI more than 1 decade earlier in comparison to those without such a history. The extent of their coronary disease is similar to the older patients without a positive FH. The better prognosis of patients with a positive FH is mostly explained by their younger age. PMID- 15334024 TI - Aortic valve stenosis: fatal natural history despite normal left ventricular function and low invasive peak-to-peak pressure gradients. AB - For many years, the severity of valvular aortic stenosis (AS) was evaluated mainly on the basis of cardiac catheterization. In many centers, the handy peak to-peak transvalvular pressure difference or 'peak-to-peak gradient' in relation to left ventricular function was used as a crucial feature in taking a decision regarding valve substitution. In a prospective study during the period 1994-1997, 150 consecutive patients with AS were examined systematically using cardiac catherization as well as transthoracic (TTE) and transesophageal echocardiography. The study was performed in order to compare the diagnostic accuracy and reproducibility of the three modalities with the purpose of improving our evaluation strategy. We found that the three methods were able to determine the aortic valve area with similar accuracy and reproducibility. The data thus support earlier papers and the currently recommended strategy of managing most patients on the basis of TTE since this approach is more rapid and gentle to the patients. In accordance with the past policy of our department, however, considerable weight was put on the invasive data during the study period. Thus, 12 patients with invasive peak-to-peak gradient <50 mm Hg and no severe depression of left ventricular function were not offered valve replacement, despite symptoms and significant valve area reductions. At 2.5 years of follow-up, 6 had died, 3 of severe heart failure, 2 while awaiting scheduled valve replacement, and 1 during aortocoronary bypass surgery. Another 3 patients later experienced further symptom progression and underwent successful aortic valve replacement. In the remaining 3 patients, all free from coronary stenoses and other valvular heart disease than AS, heart failure symptoms had worsened considerably during continued medical therapy. In conclusion, we do not recommend consideration of the peak-to-peak gradient in the process of deciding whether or not AS patients should receive valve replacement. A low peak-to-peak gradient does not exclude severe AS, even in the presence of preserved left ventricular function. PMID- 15334025 TI - Safety of transvenous temporary cardiac pacing in patients with accidental digoxin overdose and symptomatic bradycardia. AB - BACKGROUND: Patients with digoxin intoxication may need transvenous temporary cardiac pacing (TCP) when symptomatic bradyarrhythmias are present. However, it has been reported that TCP might be associated with fatal arrhythmias in patients with acute digitalis intoxication caused by attempted suicide. The aim of this study was to assess the safety of TCP in patients with accidental digoxin-related symptomatic bradyarrhythmias. MATERIALS AND METHODS: Seventy patients (30 men; age 74 +/- 12 years) were enrolled in this retrospective study. Patients were divided into two groups: group 1 with TCP and group 2 without TCP. A digoxin overdose was defined as a serum digoxin level higher than 2.0 ng/ml combined with the presence of digoxin-related symptoms. Detailed clinical characteristics were reviewed on the basis of the medical records. RESULTS: Group 1 included 24 patients (34.3%, 10 men). The rhythms prior to pacemaker insertion in group 1 included sinus arrest with junctional bradyarrhythmias (n = 9), atrial fibrillation with a slow ventricular rate (n = 11), and high-degree atrioventricular block (n = 4). The mean duration of pacemaker implantation was 5.8 +/- 2.9 days (2-12 days). There was no major arrhythmic event or mortality after TCP in group 1. Two patients in group 2 (4%) died of ventricular tachyarrhythmias. Group 1 had a higher level of blood urea nitrogen (45.1 +/- 26.0 vs. 33.4 +/- 19.3 mg/dl), of left ventricular ejection fraction (68 vs. 56%), and of digoxin (4.4 +/- 2.1 vs. 3.4 +/- 1.3 ng/ml) but a lower serum calcium level (8.7 +/- 0.6 vs. 9.1 +/- 0.8 mg/dl). CONCLUSION: TCP was safe for patients with a digoxin overdose complicated by symptomatic bradycardia and should be recommended in such situations. However, this conclusion does not apply to acute digoxin intoxication as a result of attempted suicide. PMID- 15334026 TI - Composite valve graft versus separate aortic valve and ascending aortic replacement. AB - To ascertain if the operative technique has any influence on outcome, the surgical results after aortic root replacement using either a composite valve graft (CVG) or a separate graft and valve (SVG) were analyzed. Eighty-four patients received a CVG, and 36 had SVG replacement. The operative mortality rate was 6% for patients receiving a CVG and 3% for SVG replacement (nonsignificant). Follow-up extended to 21 years (mean 124 +/- 45 months). The type of the procedure (SVG versus CVG) was not a significant predictor of in-hospital mortality, length of hospital stay, subsequent root dilatation (SVG), anastomotic dehiscence and subsequent surgery. The early and long-term results after CVG or SVG were similar, which reflects proper patient selection. PMID- 15334027 TI - Tolerability of beta-blocker initiation and titration with bisoprolol and carvedilol in congestive heart failure -- a randomized comparison. AB - In clinical trials beta-blockers (BB) are well tolerated in patients with systolic congestive heart failure (CHF). In contrast, in daily practice treatment initiation and titration appear to be more difficult and may differ in various BB -- but systematic data are lacking. We randomized 87 patients with systolic CHF (mean age 70.1 +/- 10.6 years, 24% females, LVEF 0.28 +/- 0.10, NYHA III-IV 29%) to treatment with either bisoprolol or carvedilol, in order to assess and compare the maximally achievable number of patients on treatment with these BB and the maximally achieved dosage during a minimum of 2 months attendance at our out patient heart failure clinic. In those randomized to bisoprolol, mean dose at 2 months was 33% of target dose (3.1 +/- 2.6 mg) and 41% at discharge. In those on carvedilol, the mean dose at 2 months was 27% of target dose (13.4 +/- 14.0 mg) and 32% at discharge. Thirty-nine and 40% of the bisoprolol and carvedilol treated patients, respectively, had stopped treatment at discharge. None of the figures differed significantly between the two treatment groups and no baseline parameter predicted BB tolerability. Thus, almost twice the number of unselected patients with systolic CHF attending a heart failure clinic focusing on systematic medical titration were intolerant to BB treatment and only a minority reached target dose treatment. The present data reveal no clear difference with respect to tolerability of carvedilol and bisoprolol, and no parameter predicts tolerability. PMID- 15334028 TI - Effect of withdrawal of statin on C-reactive protein. AB - BACKGROUND: C-reactive protein is considered a risk factor for coronary artery disease. In addition to its lipid-lowering properties, statin decreases the level of C-reactive protein. Abrupt cessation of statin therapy during treatment could increase the incidence of cardiac events in patients with atherosclerotic heart disease. The changes of C-reactive protein after withdrawal of statin therapy are still unknown. METHODS: Twenty patients with hyperlipidemia received statin (atorvastatin, 10 mg/day) therapy for 3 months. The levels of lipid profiles and C-reactive protein were assessed before receiving the statin therapy, immediately after 3 months of therapy, and on the 3 consecutive days after withdrawal of statin treatment. RESULTS: After 3 months of statin therapy, the total cholesterol, low-density lipoprotein cholesterol (LDL-chol), and C-reactive protein were significantly reduced (264.94 +/- 16.23 vs. 183.44 +/- 16.34 mg/dl, 183.17 +/- 34.56 vs. 122.00 +/- 17.66 mg/dl, and 2,309.00 +/- 437.85 vs. 1,257.95 +/- 207.99 ng/ml, respectively). The level of C-reactive protein increased on the second day after withdrawal of statin therapy (2,590.14 +/- 1,045.05 vs. 1,257.95 +/- 207.99 ng/ml); however, the total cholesterol and LDL-chol did not increase during the 3-day period after withdrawal of statin therapy. CONCLUSIONS: The increase in the level of C-reactive protein after withdrawal of statin therapy may be a contributing factor to the increased incidence of cardiac events in patients who have abruptly stopped statin therapy. PMID- 15334029 TI - What have the new definition of acute myocardial infarction and the introduction of troponin measurement done to the coronary care unit? Impacts on admission rate, length of stay, case mix and mortality. AB - OBJECTIVE: To assess the impact of the new American College of Cardiology/European Society of Cardiology definition of acute myocardial infarction (AMI) and the introduction of troponin measurement on the coronary care unit (CCU). METHODS: This was a retrospective cohort study performed in a tertiary care university hospital. All admissions to the CCU during the year before (period 1, year 2000, n = 1,134) and the year after (period 2, year 2002, n = 1,360) the introduction of troponin measurement and the new AMI definition were studied. We studied baseline characteristics, case load, distribution of admission diagnoses, management and outcome of patients in the two periods. RESULTS: There was a 20% increase in the number of CCU admissions, driven solely by a 141% increase in the burden of non-ST elevation AMI (NSTEMI) (p < 0.01). This increase was not a mere reflection of a change in diagnostic criteria, as the overall burden of non-ST elevation acute coronary syndromes (ACS) (NSTEMI + unstable angina) increased by 46%, suggesting referral of many more patients to the CCU. Despite a 42% increase in the number of angiograms performed, the proportion of ACS patients who had an angiogram declined. AMI patients in period 2 were older and had higher rates of coronary risk factors but had a higher chance of receiving a guideline-based therapy. Length of CCU stay decreased by a whole day for all ACS patients. 30-day mortality for AMI patients did not change significantly. CONCLUSIONS: The new AMI definition had a dramatic impact on the CCU case load, case mix and length of stay and on the ability to provide early coronary angiography. PMID- 15334030 TI - A novel role for vernix caseosa as a skin cleanser. AB - OBJECTIVES: Skin cleansing is a complex process involving endogenous and exogenous mechanisms. This study examines the role of vernix caseosa in the process of skin cleansing in the perinatal period. METHODS: Vernix was evaluated as an exogenously applied skin cleanser using digitized image analysis which quantified residual carbon particles following a standardized cleansing assay. In addition, the detachment of vernix from human cadaveric skin and Gore-Tex supports was investigated following timed exposures to a variety of commonly used commercial surfactants. Detachment was quantified spectrophotometrically as increased turbidity at 650 nm. RESULTS: Image analysis showed that exogenous application of vernix exhibited a cleansing capability comparable or superior to standard skin cleansers. Dose-dependent increases in solution turbidity (vernix detachment) were seen following exposure of vernix-covered Gore-Tex vehicles to sodium laureth sulfate, sodium lauryl sulfate, and cocamidopropyl betaine solutions. Similar results were seen with cadaveric skin. CONCLUSIONS: These results demonstrate a role for vernix caseosa as a skin cleanser. Previous views of vernix as a soil or skin contaminant at birth need to be reevaluated. PMID- 15334031 TI - Erythropoietin increases glutathione peroxidase enzyme activity and decreases lipid peroxidation levels in hypoxic-ischemic brain injury in neonatal rats. AB - BACKGROUND: We have previously shown that erythropoietin (Epo) exerts neuroprotective effects in the Rice-Vannucci model of neonatal hypoxic-ischemic brain injury. However, the mechanisms of Epo protection in this model are still unclear. OBJECTIVES: In the present study, we studied the effects of systemically administered Epo on lipid peroxidation levels and antioxidant enzyme (superoxide dismutase and glutathione peroxidase) activities following hypoxic-ischemic brain injury in neonatal rats. METHODS: Seven-day-old Wistar rat pups were subjected to left carotid artery occlusion followed by 2.5 h of hypoxic exposure. Brain lipid peroxidation levels and antioxidant enzyme activities were measured in the injured hemispheres 24 h after the hypoxic-ischemic insult. RESULTS: Hypoxic ischemic injury significantly increased the thiobarbituric acid-reactive substance levels in the injured hemispheres as compared to the control group. In addition, glutathione peroxidase activity was significantly elevated in Epo treated animals compared to saline-treated animals and the control group. CONCLUSIONS: These results suggest that Epo exerts neuroprotective effects against hypoxic-ischemic brain injury at least partially via the modulation of antioxidant enzyme activity. PMID- 15334032 TI - Effects of long-term intervention with Lactobacillus helveticus-fermented milk on bone mineral density and bone mineral content in growing rats. AB - BACKGROUND: Lactobacillus helveticus-fermented milk has been shown to increase calcium absorption compared to ordinary sour milk. In the present study the possible effect of L. helveticus-fermented milk on bone was studied in growing rats. METHODS: Spontaneously hypertensive male rats, which develop an osteoporotic bone disorder with age, were randomized into 5 groups (n = 10) receiving milk fermented with L. helveticus 16H and, as reference drinks, milk fermented with L. helveticus and Saccharomyces cerevisiae, sour milk, skim milk or water, for 14 weeks. After the intervention bone mineral density and bone mineral content were measured by dual-energy X-ray absorptiometry. The femur weight, length and volume were measured before ashing. From the ashes the weight and mineral content were assessed. RESULTS: As the body weight gain differed significantly between the groups, the results were related to the body weight. The L. helveticus-fermented milk intervention significantly increased the bone mineral density and bone mineral content compared to the sour milk, skim milk and water interventions. The mean values of the bone mineral density and bone mineral content were higher in the L. helveticus-fermented milk group compared to the Saccharomyces-fermented milk group but the difference was not statistically significant. CONCLUSION: L. helveticus-fermented milk increases bone mineral density and bone mineral content in relation to body weight in the long-term feeding of growing rats. The mechanism of L. helveticus remains to be discovered. PMID- 15334034 TI - Adverse-effect profile of kava. AB - The use of alternative therapies has increased substantially over the last decade, particularly for more chronic conditions such as anxiety. Among the most widely used treatments are medicinal herbs, or phytomedicines, such as kava (Piper methysticum), which has demonstrated anxiolytic activity in both animal models and clinical samples. Kava has several advantages over conventional pharmacologic treatments for anxiety--in clinical settings it has been associated with better tolerability and lack of physiologic dependence and withdrawal. However, phytomedicines are not rigorously regulated in the United States and systematically collected safety data are very limited. These issues are a leading concern regarding the safety of medicinal herbs such as kava. In this report, the safety profile for kava is provided, including findings from a study of its use in generalized anxiety disorder. Safety parameters assessed include occurrence of adverse events, withdrawal symptoms, effect on heart rate, blood pressure, laboratory assessments, and sexual function. No differences were found between kava and placebo on any of the parameters evaluated. The data support the safety of kava in treating anxiety at 280 mg kava lactones/day for 4 weeks. PMID- 15334033 TI - Differential effects of cooked common bean (Phaseolus vulgaris) and lentil (Lens esculenta puyensis) feeding on protein and nucleic acid contents in intestines, liver and muscles in rats. AB - AIM: Our aim was to investigate the influence of legume feeding on the protein and nucleic acid content of intestinal tissues and muscles. METHODS: Growing male Wistar rats were fed ad libitum on balanced diets containing cooked common bean or lentil as the unique protein source (180 g.kg(-1) dry matter) for 20 days. The control group was pair-fed with an iso-energetic, iso-nitrogenous balanced casein diet. RESULTS: Intestinal tissues were heavier in the legume-fed groups and higher relative mass (g per 100 g body mass) of protein, RNA and DNA were found in these tissues as compared to those of the control groups. In liver protein and RNA masses were significantly lower in the bean group than in the control group but the DNA content was not different in the legume and in the control groups. In gastrocnemius and soleus muscles, there was no significant effect of legume feeding on the fresh mass and on the protein and RNA contents, but the DNA content of the m. gastrocnemius was significantly lower in both legume groups than in their control group. The ribosomal capacity of intestines, liver and muscles was not significantly affected by legume feeding. CONCLUSION: Legume feeding had a trophic effect on both proximal and distal intestinal tissues; dietary fiber appears to be the main cause of this effect. PMID- 15334035 TI - Alternative, complementary, or not: is mainstream medicine ready for herbal medicine? PMID- 15334036 TI - Evaluating medicinal herbs. PMID- 15334037 TI - Herb-drug interactions: focus on pharmacokinetics. AB - Recent literature regarding drug-drug, herb-drug, and food-drug interactions must not be ignored; nor can they always be taken at face value. Studies have shown that St. John's wort (SJW) (Hypericum perforatum) can reduce plasma levels of indinavir, cyclosporin, digoxin, and possibly other drugs as well. Current knowledge regarding the metabolism of these medications suggests that the cytochrome P450 (CYP) drug metabolizing enzyme systems cannot account for all these effects. It has been reported that the P-glycoprotein (Pgp) transmembrane pump is also induced by SJW. Medications that are substrates of both CYP 3A4 and Pgp are of particular concern and may pose special interaction risks when combined with certain foods or botanical products such as SJW. PMID- 15334038 TI - St. John's Wort: clinical status in psychiatry. AB - Depression and anxiety are among the top 10 health problems for which complementary and alternative therapies (CATs) are most frequently used, and medicinal herbs are among the most popular of these treatments. St. John's wort (Hypericum perforatum) is a perennial herb that has become a widely used depression therapy. Extracts of hypericum have shown affinity for receptors within multiple neurochemical systems. The primary active substance responsible for the antidepressant effect is not well defined, but most work has concentrated specifically on the hypericin and hyperforin components. Although hypericum has demonstrated significant antidepressant and antianxiety effects in multiple studies, there are several recent studies that do not support the previous evidence. In all reported studies, hypericum extracts have been well tolerated. In addition, new psychiatric uses for hypericum in obsessive-compulsive disorder, generalized anxiety disorder, menopausal symptoms, and alcohol dependence have been reported. Because patients are choosing to pursue CAT as a first-line therapy, psychiatrists will need to have a better understanding of phytomedicines used for treating depression and anxiety, and thus be better prepared to serve as effective allies of their patients. PMID- 15334039 TI - The use of valerian in neuropsychiatry. AB - Valerian is a medicinal agent deriving from the plant Valeriana officinalis L. We reviewed the available literature on the use of valerian preparations in the treatment of neuropsychiatric disorders. Preclinical studies suggest that valerian has sedative and muscle-relaxant effects. Few clinical trials with valerian have been carried out in conditions other than insomnia. The insomnia studies have methodologic shortcomings but suggest that some preparations lead to significant subjective improvement in sleep complaints with remarkably few side effects. Furthermore, some evidence indicates that valerian preparations may have a mechanism of action and clinical characteristics that differ from the benzodiazepine-related sedative/hypnotics, making them more suitable for long term use. If this safety profile and the plant's sedative/hypnotic efficacy are confirmed in double-blind, placebo-controlled trials with carefully and consistently prepared valerian compounds, then those compounds would fill an important and presently unfilled niche in the treatment of insomnia. PMID- 15334040 TI - The self-assessment of resilience and anxiety: psychometric properties. AB - This report describes the psychometric properties of the Self-Assessment of Resilience and Anxiety (SARA) scale within the context of a study of kava for generalized anxiety. This eight-item, patient-rated scale includes questions designed to measure calmness, mental clarity, confidence, physical well being, sociability, and resilience. The SARA scale was administered during a clinical trial of kava versus placebo in 38 subjects diagnosed with generalized anxiety disorder (GAD). Validation of the SARA scale was assessed against the Hamilton Rating Scale for Anxiety, Hospital Anxiety and Depression Scale, Penn State Worry Questionnaire, Sheehan Disability Scale, Arizona Sexual Experience scale, and Clinical Global Impression of Illness Improvement scale. Good test-retest reliability, internal consistency, convergent validity, and sensitivity to treatment were identified, and factor analyses revealed a three-factor internal structure. The SARA scale was responsive to symptom change over time and correlated with comparable GAD symptom measures. The SARA scale demonstrates solid psychometric properties and may serve as a reliable and valid measure in the use of kava or related medicinal herbs. PMID- 15334041 TI - Sleep apnea and congestive heart failure. AB - Sleep apnea encompasses 2 forms of sleep disordered breathing, namely obstructive and central sleep apnea. Both these conditions are prevalent in patients with congestive heart failure (CHF) despite quite different etiology and pathogenesis. The last 15 years have seen the development of a large database of mechanistic data implicating both these conditions in the progression of cardiac dysfunction in patients with heart failure. Epidemiological data have also revealed that obstructive sleep apnea may be an independent risk factor for the development of cardiac diseases. Central sleep apnea, conversely, is more likely to emerge as a consequence of severe cardiac dysfunction, but through an elaborate vicious cycle could potentially lead to augmentation of sympathetic activity and contribute to further cardiac decline. In recent years a number of randomized controlled trials suggests secondary endpoints such as symptoms, sympatho-excitation and left ventricular function can be improved with the effective therapies available for both central and obstructive sleep apnea in patients in which these conditions co exist. Mortality data is emerging also, and the first of a large scale mortality trial assessing the effect of attenuating central sleep apnea with continuous positive airway pressure in patients with moderate to severe CHF, is well underway. This review summarizes the important mechanistic, epidemiological and interventional studies in relation to sleep apnea and congestive heart failure with some commentary on the future direction of this rapidly growing field. PMID- 15334042 TI - Sleep apnea and hypertension. AB - Obstructive sleep apnea (OSA) is the most common form of sleep-disordered breathing and frequently coexists with obesity. Almost 15 million Americans are affected by this disorder. This prevalence is likely increasing, given the current epidemic of obesity. Recent data confirm an association between sleep apnea and several cardiovascular disease conditions, suggesting that OSA may be a new risk factor for coronary artery disease, heart failure, heart rhythm disturbances and hypertension, independent of body mass index. In this review, the authors focus on the nature of the association between OSA and hypertension, the evidence suggesting a causal interaction, and discuss the potential pathophysiologic mechanisms responsible. These mechanisms include activation of the sympathetic and renin-angiotensin-aldosterone systems (RAAS), oxidative stress, and systemic and vascular inflammation, all of which could link OSA to a sustained increase in blood pressure. The authors also review potential therapeutic strategies for the hypertensive patient with OSA. PMID- 15334043 TI - Is sleep apnea a risk factor for stroke? A critical analysis. AB - Stroke is the 3rd leading cause of death and a major cause of serious long-term disability in the United States. There are several well established and modifiable risk factors for the development of stroke. These include arterial hypertension, cardiac disease, dyslipidemia, diabetes mellitus and smoking among others. Sleep apnea has been found at alarmingly high rates (>50%) in patients with acute stroke as well as after neurologic recovery leading some to speculate that sleep apnea had been present prior to stroke. Sleep apnea is highly prevalent in the general population with a frequency of 2% to 4%. Sleep apnea is associated with high incidence of obesity, coronary artery disease and hypertension. There are several hematologic and hemodynamic changes in sleep apnea that can play significant roles in the pathogenesis of stroke. In this review, the author provides a critical analysis of the association between sleep apnea and stroke. There is convincing evidence to believe that sleep apnea is a modifiable risk factor for stroke, however, prospective studies are needed to establish the cause-and-effect relationship. Stroke and sleep-related breathing disorders are both common and are associated with significant morbidity and mortality. Several recent large epidemiological studies have shown a strong association between these 2 disorders independent of known risk factors for stroke. Understanding the link between obstructive sleep apnea and stroke may provide a novel preventative and therapeutic approach in the management of stroke. PMID- 15334044 TI - Sleep disordered breathing in patients with chronic obstructive pulmonary disease. AB - Sleep has effects on breathing, including changes in respiratory control, airways resistance and muscular contractility. These sleep-related modifications in the respiratory system do not induce adverse effects in healthy subjects, but may cause problems in patients with chronic obstructive pulmonary disease (COPD). Hypo-ventilation causes the most important gas-exchange alteration during sleep in COPD patients, leading to hypercapnia and hypoxemia, especially during rapid eye-movement (REM) sleep. Blood gases alterations lead to increased arousals, sleep disruption, pulmonary hypertension and higher mortality. The presence of other sleep-related breathing disorders, like sleep apnea syndrome, may induce a more pronounced impairment of gas exchange, both during sleep and wakefulness, and development of symptoms like excessive daytime somnolence. Nocturnal oximetry is recommended to evaluate gas exchange during sleep in COPD patients. Sleep studies are usually indicated when there is a possibility of sleep apnea or obesity-hypoventilation syndrome. The role of non-invasive mechanical ventilation in managing COPD patients with nocturnal hypoventilation is discussed. PMID- 15334045 TI - Treatment of sleep disordered breathing and obstructive sleep apnea. AB - The management of sleep disordered breathing (SDB) and obstructive sleep apnea (OSA) in adults can be challenging. Treatment recommendations are based on a number of factors including the severity of SDB, the existence and extent of comorbid conditions, the severity of presenting symptoms and patient preference. General management includes addressing lifestyle issues, particularly weight loss and the avoidance of sedatives. The primary specific treatment modality for SDB is continuous positive airway pressure (CPAP). However, for selected patients that cannot accept this therapy, other modalities such as upper airway surgery and oral appliances should be considered. PMID- 15334046 TI - Laser-assisted uvulopalatoplasty in the management of snoring and obstructive sleep apnea syndrome. AB - Laser-assisted uvulopalatoplasty (LAUP) was initially proposed as an alternative method to treat habitual snoring. Throughout the years the indications for LAUP have broadened, however, it has also remained an area of controversy. LAUP is a staged office-based procedure involving removal of excessive uvular mucosa and creation of transpalatal vertical troughs to effectively widen the retropalatal airway. Since postlaser treatment mimics the palatal appearance after uvulopalatopharyngoplasty (UPPP), similar surgical outcomes can be expected in properly selected patients. Although still an area of controversy, the medical literature has supported the cost effectiveness, safety, and comparable efficacy to the standard of UPPP in the treatment of snoring and obstructive sleep apnea syndrome (OSAS). In select patients, LAUP is a viable option for the management of not only snoring, but also OSAS. PMID- 15334047 TI - Cytokines alone for PBPC collection in patients with advanced gynaecological malignancies: G-CSF vs G-CSF plus EPO. PMID- 15334048 TI - Transmission of donor illness by stem cell transplantation: should screening be different in older donors? AB - With increasing donor age, the potential of transmitting diseases from donor to recipient reaches new dimensions. Potentially transmittable diseases from donors include infections, congenital disorders, and acquired illnesses like autoimmune diseases or malignancies of hematological or nonhematological origin. While established nonmalignant or malignant diseases might be easy to discover, early stage hematological diseases like CML, light-chain multiple myelomas, aleukemic leukemias, occult myelodysplastic syndromes and other malignant and nonmalignant diseases might not be detectable by routine screening but only by invasive, new and/or expensive diagnostic tests. In the following article, we propose recommendations for donor work-up, taking into consideration the age of the donors. In contrast to blood transfusions, stem cells from donors with abnormal findings might still be acceptable for HCT, when no other options are available and life expectancy is limited. This issue is discussed in detail in relation to the available donor and stem cell source. Finally, the recommendations presented here aim at harmonized worldwide work-up for donors to insure high standard quality. PMID- 15334049 TI - Results of autologous transplantation in lymphoma are not improved by increasing the dose of etoposide in the BEAM regimen: a single-centre sequential-cohort study. AB - We have undertaken a retrospective sequential-cohort analysis of 131 lymphoma patients treated with the BEAM regimen and autologous stem cell transplantation, to compare BEAM at standard doses (sBEAM; n = 67 from May 1990 to April 1995) and BEAM with escalated etoposide dose from 800 to 1600 mg/m(2) (eBEAM; n = 64 from May 1995 to June 1999). Transplant-related mortality and incidence of secondary malignancies were similar in both groups. Disease progression was significantly lower in indolent lymphoma (IL) patients receiving eBEAM (7 vs 43%), although survival was comparable due to a higher toxic mortality in the eBEAM group. The 5 year event-free survival and overall survival were better in Hodgkin's disease (HD) patients treated with eBEAM (70 and 77%, respectively) compared to sBEAM (58 and 69%, respectively), but the difference was not statistically significant. In aggressive lymphomas, no difference was detected between groups. Our results indicate that while escalation of the etoposide doses in the BEAM conditioning regimen does not appear to improve outcome, encouraging results in IL and HD may warrant further studies. PMID- 15334050 TI - Infusion of unwashed umbilical cord blood stem cells after thawing for allogeneic transplantation. PMID- 15334051 TI - Commentary on 2003-808: infusion of unwashed umbilical cord blood stem cells after thawing for allogeneic transplantation. PMID- 15334052 TI - Imatinib mesylate (Glivec) pre-treatment does not have a negative effect on outcome of allogenic hematopoietic stem cell transplantation in Philadelphia positive leukemias. PMID- 15334053 TI - Identification of SCN3B as a novel p53-inducible proapoptotic gene. AB - Tumor suppressor p53 is a transcription factor that induces growth arrest and/or apoptosis in response to cellular stress. To identify novel p53-inducible genes, we compared the expression of genes in normal mouse embryo fibroblasts (MEFs) to p53-null cells by cDNA representational difference analysis. We report here that expression of endogenous sodium channel subunit beta 3 (SCN3B) is upregulated in mouse embryonic fibroblasts by DNA damage in a p53-dependent manner. In addition, we found that SCN3B levels are upregulated in human cancer cell lines by DNA damaging agents, as well as by overexpression of p53, but not significantly by p63 or p73. Furthermore, we identified two putative p53-binding sites upstream of the first exon (RE1) and in the third intron (RE2). The p53 protein can directly interact with the putative p53-binding sites in vivo, as assessed by chromatin immunoprecipitation. A reporter gene assay revealed that these two p53-binding sites are functional response elements. The SCN3B protein appears to be localized to the endoplasmic reticulum (ER). Introduction of the SCN3B gene into T98G and Saos2 cells potently suppressed colony formation. Furthermore, we found that adenovirus-mediated transfer of SCN3B induced apoptosis when combined with anticancer agents. The results presented here suggest that SCN3B mediates a p53 dependent apoptotic pathway and may be a candidate for gene therapy combined with anticancer drugs. PMID- 15334054 TI - Hepatitis C viral proteins interact with Smad3 and differentially regulate TGF beta/Smad3-mediated transcriptional activation. AB - Transforming growth factor-beta (TGF-beta) is a pleiotropic cytokine implicated as a pathogenic mediator in various liver diseases. Enhanced TGF-beta production and lack of TGF-beta responses are often observed during hepatitis C virus (HCV) infection. In this study, we demonstrate that TGF-beta-mediated transactivation is decreased in cells exogenously expressing the intact HCV polyprotein. Among 10 viral products of HCV, only core and nonstructural protein 3 (NS3) physically interact with the MH1 (Mad homology 1) region of the Smad3 and block TGF beta/Smad3-mediated transcriptional activation through interference with the DNA binding ability of Smad3, not the nuclear translocation. However, the interactive domain of NS3 extends to the MH2 (Mad homology 2) region of Smad3 and a distinction is found between effects mediated, respectively, by these two viral proteins. HCV core, in the presence or absence of TGF-beta, has a stronger suppressive effect on the DNA-binding and transactivation ability of Smad3 than NS3. Although HCV core, NS3, and the HCV subgenomic replicon all attenuate TGF beta/Smad3-mediated apoptosis, only HCV core represses TGF-beta-induced G1 phase arrest through downregulation of the TGF-beta-induced p21 promoter activation. Along with this, HCV core, rather than NS3, exhibits a significant inhibitory effect on the binding of Smad3/Sp1 complex to the proximal p21 promoter in response to TGF-beta. In conclusion, HCV viral proteins interact with the TGF beta signaling mediator Smad3 and differentially impair TGF-beta/Smad3-mediated transactivation and growth inhibition. This functional counteraction of TGF-beta responses provides insights into possible mechanisms, whereby the HCV oncogenic proteins antagonize the host defenses during hepatocarcinogenesis. PMID- 15334055 TI - The cyclolignan PPP induces activation loop-specific inhibition of tyrosine phosphorylation of the insulin-like growth factor-1 receptor. Link to the phosphatidyl inositol-3 kinase/Akt apoptotic pathway. AB - The insulin-like growth factor-1 receptor (IGF-1R) is crucial for many functions in neoplastic cells, for example, antiapoptosis. Recently, we demonstrated that the cyclolignan PPP efficiently inhibited phosphorylation of IGF-1R without interfering with insulin receptor activity. PPP preferentially reduced phosphorylated Akt, as compared to phosphorylated Erk1/2, and caused apoptosis. Now, we aimed to investigate how PPP inhibits the IGF-1R tyrosine kinase (IGF 1RTK) and the PI3K/Akt apoptotic pathway. Using a baculovirus driven IGF-1RTK we found that PPP interfered with tyrosine phosphorylation in the activation loop of the kinase domain. Specifically, it blocked phosphorylation of tyrosine (Y) 1136, while sparing the two others (Y1131 and Y1135). To explore the impact of inhibition of Y1136 on Akt phosphorylation we transfected P6 cells (overexpressing IGF-1R) and malignant melanoma cells with different IGF-1R mutants, including Y1136F (tyrosine replaced by phenylalanine). Y1136F was found to strongly decrease IGF-1 stimulated phosphorylation of Akt. Conversely, Akt phosphorylation was weakly affected in the Y1131F transfectant. Taken together, our data suggest that the preferential inhibition of phosphorylated Akt, after PPP treatment, may be due to specific inhibition of Y1136. PPP was proven not to interfere directly with Akt or any of its downstream molecules in the apoptotic pathway. PMID- 15334056 TI - Transformation suppressor protein Pdcd4 interferes with JNK-mediated phosphorylation of c-Jun and recruitment of the coactivator p300 by c-Jun. AB - The transformation suppressor gene Pdcd4 (programmed cell death gene 4) inhibits the tumor-promoter mediated transformation of mouse keratinocytes and has recently been implicated as a potential tumor suppressor gene in the development of human lung cancer. Biochemical analysis has suggested that the Pdcd4 protein is involved in protein translation as well as in nuclear events. Recent work has shown that Pdcd4 suppresses the transactivation of AP-1 responsive promoters by c Jun, suggesting that the transformation-suppressor activity of Pdcd4 might be due, at least in part, to the inhibition of c-Jun activity. Here, we have addressed how Pdcd4 inhibits c-Jun. We show that Pdcd4 interferes with the phosphorylation of c-Jun by Jun N-terminal kinase (JNK). The inhibition of c-Jun phosphorylation by Pdcd4 appears not to be due to a general suppression of JNK activity, our data rather suggest that Pdcd4 interacts with c-Jun and thereby blocks phosphorylation of c-Jun. In addition to affecting c-Jun phosphorylation, Pdcd4 blocks the recruitment of the coactivator p300 by c-Jun. Taken together, our results strongly suggest that Pdcd4 is directly involved in the regulation of c-Jun activity. PMID- 15334057 TI - MRP-1/CD9 gene transduction downregulates Wnt signal pathways. AB - Motility-related protein-1 (MRP-1/CD9) is a transmembrane glycoprotein that has been implicated in cell adhesion, motility, proliferation, and differentiation. It has a functional role as a tumor metastatic suppressor. During tumor progression, a reduction of MRP-1/CD9 gene expression results in tumor cells with a high metastatic potential. However, the mechanism of action of MRP-1/CD9 is still unclear. We studied changes of gene expression in relation to MRP-1/CD9 gene transduction into tumor cell lines, HT1080 and A549, using microarray assays and real-time PCR. Consequently, we have demonstrated that MRP-1/CD9 gene transduction can downregulate expression of several Wnt family genes, such as Wnt1, Wnt2b1 and Wnt5a, and their target genes, including WISP-1 (Wnt-1 induced secreted protein 1), WISP-3, c-Myc, vascular endothelial growth factor-A, and matrix metalloproteinase-26. Western blot analyses also showed that MRP-1/CD9 gene transduction downregulated expression of Wnt1 protein and its target proteins. In addition, a neutralizing anti-MRP-1/CD9 monoclonal antibody inhibited the downregulation of Wnt signal pathways in MRP-1/CD9-transfected cells. The present study has revealed that the MRP-1/CD9 signal is located upstream of the Wnt signal pathways. Therefore, MRP-1/CD9 could suppress cell transformation including epithelial to mesenchymal transition through downregulation of Wnt1, and might suppress tumor metastasis through downregulation of Wnt5a. PMID- 15334058 TI - Caveolin-1 inhibits breast cancer growth and metastasis. AB - Caveolin-1 was identified in a screen for genes involved in breast cancer progression. Caveolin-1 is the major protein component of caveolae, flask-shaped invaginations found in a number of different cell types. Using an orthotopic model of spontaneous breast cancer metastasis, caveolin-1 was found to be expressed in low and non-metastatic primary tumors, but at much lower levels in highly metastatic 4T1.2 and 4T1.13 tumors. Exogenous expression of caveolin-1 at moderate levels in 4T1.2 cells was sufficient to suppress primary tumor growth after inoculation of cells into the mammary gland. Expression of high levels of caveolin-1 also inhibited subsequent metastasis to distant organs. Cells expressing high levels of caveolin-1 showed reduced capacity to invade Matrigel, diminished response to laminin-1 stimulation and decreased metastasis to lung and bone. This study provides the first functional evidence that caveolin-1 regulates primary breast tumor growth and spontaneous metastasis of breast cancer. PMID- 15334059 TI - Identification of an epigenetically silenced gene, RFX1, in human glioma cells using restriction landmark genomic scanning. AB - To identify the CpG islands differentially methylated in human glioma, we performed restriction landmark genomic scanning with a CpG methylation-sensitive enzyme. We found 12 spots, the intensity of which was entirely lost or decreased in both the human glioma tissues examined as compared with that in matched normal lymphocytes, indicating aberrant methylation of these CpG islands in gliomas. The expression of RFX1, one of the genes associated with the methylated CpG islands, was frequently decreased in human glioma cell lines and tissues. We also demonstrated that the isolated CpG island located in the seventh intron of the RFX1 gene had enhancer activity and was hypermethylated in all of the glioma tissues and cell lines analysed, but not in normal brains or lymphocytes. Treatment of glioma cells with a demethylating agent, 5-azacytidine, resulted in the expression of RFX1, indicating that the silencing of the RFX1 gene may be attributable to its methylation. RFX1 has been implicated in transcriptional downregulation of the proto-oncogene c-myc. By expression of the RFX1 gene, the cellular proliferative activity of glioma cells was suppressed. Taken together, these results suggest that the RFX1 gene may be epigenetically silenced in human gliomas and involved in glioma tumorigenesis. PMID- 15334060 TI - DAL-1/4.1B tumor suppressor interacts with protein arginine N-methyltransferase 3 (PRMT3) and inhibits its ability to methylate substrates in vitro and in vivo. AB - DAL-1 (differentially expressed in adenocarcinoma of the lung)/4.1B is a tumor suppressor gene on human chromosome 18p11.3 whose expression is lost in >50% of primary non-small-cell lung carcinomas. Based on sequence similarity, DAL-1/4.1B has been assigned to the Protein 4.1 superfamily whose members interact with plasma membrane proteins through their N-terminal FERM (4.1/Ezrin/Radixin/Moesin) domain, and cytoskeletal components via their C-terminal SAB (spectrin-actin binding) region. Using the DAL-1/4.1B FERM domain as bait for yeast two-hybrid interaction cloning, we identified protein arginine N-methyltransferase 3 (PRMT3) as a specific DAL-1/4.1B-interacting protein. PRMT3 catalyses the post translational transfer of methyl groups from S-adenosyl-L-methionine to arginine residues of proteins. Coimmunoprecipitation experiments using lung and breast cancer cell lines confirmed this interaction in mammalian cells in vivo. In vitro binding assays demonstrated that this was an interaction occurring via the C terminal catalytic core domain of PRMT3. DAL-1/4.1B was determined not to be a substrate for PRMT3-mediated methylation but its presence inhibits the in vitro methylation of a glycine-rich and arginine-rich methyl-accepting protein, GST (glutathione-S-transferase-GAR (glycine- and arginine-rich), which contains 14 'RGG' consensus methylation sites. In addition, induced expression of DAL-1/4.1B in MCF-7 breast cancer cells showed that the DAL-1/4.1B protein significantly inhibits PRMT3 methylation of cellular substrates. These findings suggest that modulation of post-translational methylation may be an important mechanism through which DAL-1/4.1B affects tumor cell growth. PMID- 15334061 TI - FLIP overexpression inhibits death receptor-induced apoptosis in malignant mesothelial cells. AB - Tumors have developed several forms of resistance to receptor-induced cell death. Here, we show that malignant mesothelial (MM) cell lines as well as primary MM cells and normal mesothelial (NM) cells express Fas and TNF-related apoptosis inducing ligand (TRAIL) receptors DR4 and DR5. We found that, although Fas expression levels are comparable, only MM cells are resistant to cell death. Furthermore, MM cells show resistance to TRAIL-induced apoptosis. Caspase-8 (FLICE) is not activated by death receptors triggering in malignant cells whereas it is well activated by nonreceptor stimuli, such as UV radiation. We found that FLIP (FLICE-Inhibitory Protein) is constitutively expressed in all MM cell lines and is more expressed in primary MM cells than in NM cells. Knockdown of FLIP expression in MM cell lines, by a FLIPsiRNA, re-established the normal response to apoptosis induced by Fas or DR4/DR5, which was blocked by pretreatment with the caspase-8 inhibitor z-IETD-fmk. These results indicate that MM cells develop an intrinsic resistance to apoptosis induced by death receptors upregulating the expression of the antiapoptotic protein c-FLIP. PMID- 15334062 TI - Oxaliplatin-induced mitochondrial apoptotic response of colon carcinoma cells does not require nuclear DNA. AB - We previously established a model of acquired oxaliplatin resistance derived from the HCT116 oxaliplatin-sensitive cell line (HCT116S) and consisting in two resistant clones (HCT116R1, HCT116R2) and their total or partial revertants (HCT116Rev1 and HCT116Rev2, respectively). Using this cellular model, we explored the contribution of mitochondrial apoptosis and nuclear DNA to oxaliplatin mediated apoptosis induction and oxaliplatin resistance. We showed that the activity of oxaliplatin is mediated by the induction of Bax/Bak-dependent mitochondrial apoptosis and that oxaliplatin resistance is mediated by a defect in Bax/Bak activation correlating with a reduced loss of the mitochondrial transmembrane potential (DeltaPsim). In addition, we observed that p53 only contributed marginally to oxaliplatin-induced cytotoxicity and was not involved in oxaliplatin resistance. Moreover and surprisingly, depletion of the nucleus in HCT116S cells did not abolish the oxaliplatin-induced DeltaPsim loss indicative of imminent apoptosis. Enucleation abolished the oxaliplatin resistance of HCT116R1 cells, while HCT116R2 cytoplasts conserved their resistant phenotype. Altogether, these data demonstrate that oxaliplatin exerts its cytotoxic effects by inducing mitochondrial apoptosis and that these effects can be initiated by interacting on other cellular structures than nuclear DNA. Resistance to oxaliplatin may imply both nuclear and cytoplasmic compartments. PMID- 15334063 TI - Luteolin sensitizes tumor necrosis factor-alpha-induced apoptosis in human tumor cells. AB - Tumor necrosis factor-alpha (TNFalpha) activates both cell death and cell survival pathways, which render most cancer cells resistant to its cytotoxicity. In this study, we found that pretreatment with luteolin, a plant flavonoid, greatly sensitized TNFalpha-induced apoptotic cell death in a number of human cancer cell lines; including colorectal cancer COLO205, HCT116 cells and cervical cancer HeLa cells. In the search of the molecular mechanisms responsible for the sensitization effect of luteolin, we discovered that luteolin inhibited TNFalpha induced activation of nuclear transcription factor-kappa B (NF-kappaB), the main survival factor in TNFalpha signaling. As a result, luteolin suppressed the expression of NF-kappaB-targeted antiapoptotic genes, including A20 and cellular inhibitor of apoptosis protein-1 (c-IAP1). The role of A20 and c-IAP1 was further confirmed by ectopic expression of these two genes, which significantly protected cell death induced by luteolin followed by TNFalpha. In addition, inhibition of NF-kappaB by luteolin led to augmentation and prolongation of c-Jun N-terminal kinase (JNK) activation induced by TNFalpha. Suppression of JNK activation, either by a synthetic JNK inhibitor (SP600125) or by overexpression of the dominant negative forms of JNK kinase 1 (JNKK1) and JNK kinase 2 (JNKK2), conferred significant protection against apoptotic cell death induced by luteolin and TNFalpha, suggesting that NF-kappaB and JNK are closely associated with the sensitization effect of luteolin. Data from this study reveal a novel function of luteolin and enhance the value of luteolin as an anticancer agent. PMID- 15334064 TI - An alternatively spliced survivin variant is positively regulated by p53 and sensitizes leukemia cells to chemotherapy. AB - Survivin is a unique member of the inhibitor of apoptosis protein family, and its expression is regulated by p53. Recent identification of several functionally divergent survivin variants augments the complexity of survivin action as well as its regulation. Here we report that survivin-2B (retaining a part of intron 2 as a cryptic exon) is positively regulated by p53, and its overexpression plays a role in sensitizing leukemia cells to chemotherapeutic drug doxorubicin. Doxorubicin treatment activated p53, downregulated survivin and survivin-DeltaEx3 but upregulated survivin-2B in EU-3, an acute lymphocytic leukemia (ALL) cell line with wild-type (wt)-p53 phenotype. In contrast, doxorubicin treatment failed to induce these alterations in EU-6 cells, a mutant-p53 ALL cell line. To specify the role of wt-p53 in regulating survivin and its variants, a temperature sensitive p53 mutant plasmid p53-143 was transfected into EU-4, a p53-null ALL cell line, to establish a subline EU-4/p53-143. When EU-4/p53-143 cell culture was shifted from 37.5 degrees C to the wt-p53-permissive temperature (32.5 degrees C), the expression of survivin and survivin-DeltaEx3 was decreased whereas survivin-2B expression was increased, confirming the distinct regulatory effect of p53 on survivin and its variants. To clarify the role of survivin-2B in the process of apoptosis, survivin-2B cDNA was cloned into pcDNA3HA vector and transfected into EU-4 cells. Enforced expression of survivin-2B in EU-4 cells inhibited cell growth and sensitized these cells to doxorubicin-induced apoptosis. These results suggest that survivin-2B variant is a proapoptotic factor and its expression is upregulated by p53. PMID- 15334065 TI - Tumor formation in p53 mutant ovaries transplanted into wild-type female hosts. AB - P53: gene alterations correlate highly with advanced ovarian carcinoma in women. In mice, p53 deficiency predominantly results in the formation of lymphomas and sarcomas. However, ovarian epithelial tumors have not been documented in p53 homozygous mutant (p53-/-) mice, probably because they die before other tumors can form. To determine whether p53-/- ovaries can develop epithelial tumors, they were transplanted into the ovarian bursae of histocompatible wild-type recipient females. The p53-/- ovarian grafts formed tumors approximately 1 year post transplantation. The tumor type was angiosarcoma, suggesting that vascular tissues are predisposed to tumor formation in p53-/-ovaries. These findings suggest that p53 deficiency alone is not sufficient for ovarian epithelial tumorigenesis in mice. Thus, other genetic lesions are likely required to develop mouse models of human ovarian cancer. PMID- 15334066 TI - Silencing of androgen-regulated genes using a fusion of AR with the PLZF transcriptional repressor. AB - The androgen receptor (AR) is a member of the nuclear receptor superfamily of ligand-activated transcription factors and plays a key role in the development and progression of prostate cancer. Current therapies include the use of antiandrogens aimed at inhibiting the transcriptional activation of AR-regulated genes by AR. Here, we explore a strategy aimed at obtaining silencing of AR regulated genes, based on the properties of the transcriptional repressor promyelocytic leukamia zinc-finger protein (PLZF). In order to do this, we have made a fusion protein between PLZF and AR, named PLZF-AR, and show that PLZF-AR is able to bring about silencing of genomically encoded AR-regulated genes and inhibit the androgen-regulated growth of LNCaP prostate cancer cells. Together, our results show that this strategy is able to bring about potent repression of AR-regulated responses and, therefore, could be of value in the development of new therapies for prostate cancer. PMID- 15334068 TI - Suppression subtractive hybridization and expression profiling identifies a unique set of genes overexpressed in non-small-cell lung cancer. AB - Expression array data for >3000 individual clones from two suppression subtractive hybridization libraries revealed 147 genes overexpressed in non-small cell lung cancer (NSCLC) cell lines. Of these 147 genes, 30 genes have previously unknown cancer association and 65 genes have been associated with cancers other than NSCLC. The identification of 52 genes previously associated with NSCLC by different methodologies supports the validity of the strategy used here. Of the 147 genes, 19 have no prior named Unigene cluster designation, and are designated herein as L1 to L19. Quantitative real-time PCR and cancer profiling arrays were used as independent validation tools to confirm tumor overexpression for five of the 'L' genes in tumor cell lines and patient samples from NSCLC and other cancers. Follow-up studies for candidate NSCLC-associated genes can be useful in providing valuable insight into the etiology of lung cancer as well as providing potentially interesting diagnostic or therapeutic targets for further investigation. PMID- 15334067 TI - Mechanistic aspects of mda-7/IL-24 cancer cell selectivity analysed via a bacterial fusion protein. AB - The human mda-7/IL-24 gene product is normally expressed in melanocytes and certain lymphocyte populations. Loss of expression, a distinctive feature of many tumor suppressor genes, has been documented at RNA and protein levels in association with melanoma progression both in vitro as well as in human tumor derived material. The MDA-7/IL-24 protein undergoes post-translational processing, including removal of an amino-terminal 48-residue signal peptide and extensive glycosylation prior to its secretion by producing cells. Its inherent cytokine properties have been documented in multiple reports, which have identified and characterized its cognate receptors and activation of the JAK/STAT signaling pathway following ligand/receptor docking. A notable and incompletely understood property of MDA-7/IL-24 is its ability to induce apoptosis in transformed cells, while having marginal growth suppressive effects on normal primary or immortalized cell lines. MDA-7/IL-24 has been delivered to cells, tumor xenografts and patients in clinical trials via a nonreplicating adenovirus (Ad.mda-7). Studies utilizing eukaryotically expressed and purified MDA-7/IL-24 protein from several sources have recapitulated some of the molecule's reported properties including receptor binding and JAK/STAT activation. Here, we report the properties and characteristics of a bacterially expressed and purified GST MDA-7 fusion protein. These studies reveal that GST-MDA-7 retains its cancer selective apoptosis-inducing properties, thereby providing a new reagent that will assist in defining the mechanism of action of this novel cytokine. In addition, retention of tumor-specific activity of GST-MDA-7 suggests that this protein may also have therapeutic applications. PMID- 15334069 TI - Nuclear localization of the adenovirus E4orf4 protein is mediated through an arginine-rich motif and correlates with cell death. AB - The adenovirus E4orf4 protein induces p53-independent death of human cancer cells by a mechanism requiring interactions with the Balpha subunit of protein phosphatase 2A. When expressed alone E4orf4 localizes predominantly in the nucleus, although significant levels are also present in the cytoplasm. While tyrosine phosphorylation of E4orf4 and recruitment of Src have been linked with E4orf4 cytoplasmic cell death functions, little is known about the functions of E4orf4 in the nucleus. In this study, we identified an arginine-rich motif (E4ARM; residues 66-75) that is necessary and sufficient for nuclear and nucleolar localization. This motif, which is highly homologous to the arginine rich nuclear and nucleolar localization motif of some lentiviral proteins, was shown to target heterologous proteins to the nucleus and to nucleoli, functions found to be dependent on the overall charge of the motif rather than on specific residues. Furthermore, mutation of arginine residues to alanines but not to lysines in E4ARM was shown to block such targeting activity and, when introduced into full-length E4orf4, to decrease induction of cell death. Finally, coexpression of the ARM motifs of E4orf4, HIV-1 Tat or Rev along with full-length E4orf4 was seen to decrease E4orf4-dependent cell killing. Thus it appears that targeting of E4orf4 to the nucleus and cell nucleoli by E4ARM is an important component of E4orf4-induced cell death. PMID- 15334070 TI - Linkage between dynamics and catalysis in a thermophilic-mesophilic enzyme pair. AB - A fundamental question is how enzymes can accelerate chemical reactions. Catalysis is not only defined by actual chemical steps, but also by enzyme structure and dynamics. To investigate the role of protein dynamics in enzymatic turnover, we measured residue-specific protein dynamics in hyperthermophilic and mesophilic homologs of adenylate kinase during catalysis. A dynamic process, the opening of the nucleotide-binding lids, was found to be rate-limiting for both enzymes as measured by NMR relaxation. Moreover, we found that the reduced catalytic activity of the hyperthermophilic enzyme at ambient temperatures is caused solely by a slower lid-opening rate. This comparative and quantitative study of activity, structure and dynamics revealed a close link between protein dynamics and catalytic turnover. PMID- 15334071 TI - Identification of the versatile scaffold protein RACK1 on the eukaryotic ribosome by cryo-EM. AB - RACK1 serves as a scaffold protein for a wide range of kinases and membrane-bound receptors. It is a WD-repeat family protein and is predicted to have a beta propeller architecture with seven blades like a Gbeta protein. Mass spectrometry studies have identified its association with the small subunit of eukaryotic ribosomes and, most recently, it has been shown to regulate initiation by recruiting protein kinase C to the 40S subunit. Here we present the results of a cryo-EM study of the 80S ribosome that positively locate RACK1 on the head region of the 40S subunit, in the immediate vicinity of the mRNA exit channel. One face of RACK1 exposes the WD-repeats as a platform for interactions with kinases and receptors. Using this platform, RACK1 can recruit other proteins to the ribosome. PMID- 15334072 TI - Tracking metastatic tumor cell extravasation with quantum dot nanocrystals and fluorescence emission-scanning microscopy. AB - Metastasis is an impediment to the development of effective cancer therapies. Our understanding of metastasis is limited by our inability to follow this process in vivo. Fluorescence microscopy offers the potential to follow cells at high resolution in living animals. Semiconductor nanocrystals, quantum dots (QDs), offer considerable advantages over organic fluorophores for this purpose. We used QDs and emission spectrum scanning multiphoton microscopy to develop a means to study extravasation in vivo. Although QD labeling shows no deleterious effects on cultured cells, concern over their potential toxicity in vivo has caused resistance toward their application to such studies. To test if effects of QD labeling emerge in vivo, tumor cells labeled with QDs were intravenously injected into mice and followed as they extravasated into lung tissue. The behavior of QD labeled tumor cells in vivo was indistinguishable from that of unlabeled cells. QDs and spectral imaging allowed the simultaneous identification of five different populations of cells using multiphoton laser excitation. Besides establishing the safety of QDs for in vivo studies, our approach permits the study of multicellular interactions in vivo. PMID- 15334073 TI - Tumor-infiltrating dendritic cell precursors recruited by a beta-defensin contribute to vasculogenesis under the influence of Vegf-A. AB - The involvement of immune mechanisms in tumor angiogenesis is unclear. Here we describe a new mechanism of tumor vasculogenesis mediated by dendritic cell (DC) precursors through the cooperation of beta-defensins and vascular endothelial growth factor-A (Vegf-A). Expression of mouse beta-defensin-29 recruited DC precursors to tumors and enhanced tumor vascularization and growth in the presence of increased Vegf-A expression. A new leukocyte population expressing DC and endothelial markers was uncovered in mouse and human ovarian carcinomas coexpressing Vegf-A and beta-defensins. Tumor-infiltrating DCs migrated to tumor vessels and independently assembled neovasculature in vivo. Bone marrow-derived DCs underwent endothelial-like differentiation ex vivo, migrated to blood vessels and promoted the growth of tumors expressing high levels of Vegf-A. We show that beta-defensins and Vegf-A cooperate to promote tumor vasculogenesis by carrying out distinct tasks: beta-defensins chemoattract DC precursors through CCR6, whereas Vegf-A primarily induces their endothelial-like specialization and migration to vessels, which is mediated by Vegf receptor-2. PMID- 15334075 TI - Impugning conflict declarations. PMID- 15334074 TI - CalDAG-GEFI integrates signaling for platelet aggregation and thrombus formation. AB - Signaling through the second messengers calcium and diacylglycerol (DAG) is a critical element in many biological systems. Integration of calcium and DAG signals has been suggested to occur primarily through protein kinase C family members, which bind both calcium and DAG. However, an alternative pathway may involve members of the CalDAG-GEF/RasGRP protein family, which have structural features (calcium-binding EF hands and DAG-binding C1 domains) that suggest they can function in calcium and DAG signal integration. To gain insight into the signaling systems that may be regulated by CalDAG-GEF/RasGRP family members, we have focused on CalDAG-GEFI, which is expressed preferentially in the brain and blood. Through genetic ablation in the mouse, we have found that CalDAG-GEFI is crucial for signal integration in platelets. Mouse platelets that lack CalDAG GEFI are severely compromised in integrin-dependent aggregation as a consequence of their inability to signal through CalDAG-GEFI to its target, the small GTPase Rap1. These results suggest that analogous signaling defects are likely to occur in the central nervous system when CalDAG-GEFI is absent or compromised in function. PMID- 15334076 TI - Viral infection and allergy. PMID- 15334078 TI - The brightening future of HIV therapeutics. PMID- 15334079 TI - Environmental conservation: bystander CD4 T cells keep CD8 memories fresh. PMID- 15334080 TI - A peek at PIAS. PMID- 15334081 TI - RPA tightens AID to DNA...editing. PMID- 15334082 TI - To help and help not. PMID- 15334084 TI - E3 ubiquitin ligases as T cell anergy factors. AB - E3 ubiquitin ligases have emerged as key molecular regulators of immune cell function. Three families of proteins with ubiquitin ligase activity have been described (the HECT, RING and U-box proteins), and each may be involved in the regulation of immune responses during infection by targeting specific inhibitory molecules for proteolytic destruction. Several HECT and RING E3 proteins have now also been linked to the induction and maintenance of immune self-tolerance: c Cbl, Cbl-b, GRAIL, Itch and Nedd4 each negatively regulate T cell growth factor production and proliferation. This review will discuss the relationship between the ubiquitination of select components of the antigen-sensing signaling apparatus in T cells and the development and maintenance of the clonal anergy state. PMID- 15334087 TI - Trigger factor in complex with the ribosome forms a molecular cradle for nascent proteins. AB - During protein biosynthesis, nascent polypeptide chains that emerge from the ribosomal exit tunnel encounter ribosome-associated chaperones, which assist their folding to the native state. Here we present a 2.7 A crystal structure of Escherichia coli trigger factor, the best-characterized chaperone of this type, together with the structure of its ribosome-binding domain in complex with the Haloarcula marismortui large ribosomal subunit. Trigger factor adopts a unique conformation resembling a crouching dragon with separated domains forming the amino-terminal ribosome-binding 'tail', the peptidyl-prolyl isomerase 'head', the carboxy-terminal 'arms' and connecting regions building up the 'back'. From its attachment point on the ribosome, trigger factor projects the extended domains over the exit of the ribosomal tunnel, creating a protected folding space where nascent polypeptides may be shielded from proteases and aggregation. This study sheds new light on our understanding of co-translational protein folding, and suggests an unexpected mechanism of action for ribosome-associated chaperones. PMID- 15334086 TI - The ubiquitin-modifying enzyme A20 is required for termination of Toll-like receptor responses. AB - A20 is a cytoplasmic protein required for the termination of tumor necrosis factor (TNF)-induced signals. We show here that mice doubly deficient in either A20 and TNF or A20 and TNF receptor 1 developed spontaneous inflammation, indicating that A20 is also critical for the regulation of TNF-independent signals in vivo. A20 was required for the termination of Toll-like receptor induced activity of the transcription factor NF-kappaB and proinflammatory gene expression in macrophages, and this function protected mice from endotoxic shock. A20 accomplished this biochemically by directly removing ubiquitin moieties from the signaling molecule TRAF6. The critical function of this deubiquitinating enzyme in the restriction of TLR signals emphasizes the importance of the regulation of ubiquitin conjugation in innate immune cells. PMID- 15334088 TI - Academic psychiatry in Poland: adjustment to rapid changes. PMID- 15334091 TI - A novel model of occlusive thrombus formation in mice. AB - A novel model to induce occlusive thrombus formation was developed in mice in vivo. Mice were simultaneously treated with ligation and cuff placement at the left carotid artery. At 7 days after the treatment, occlusive thrombus was observed at the intracuff region, but not in the distal and proximal regions of the cuff, and not induced by a single treatment of ligation or cuff placement. The plasma levels of von Willebrand factor (vWF), which represent the endothelial status, were significantly increased in combined treatment of ligation and cuff placement 1 day after the operation. Whereas no significant changes in plasma vWF were observed in either single treatment of ligation or cuff placement. The expression of vWF, considered to be the endothelial marker, was detected on the luminal surface distal and proximal to the cuff and the carotid artery in the single treatment groups treated with either ligation or cuff placement, but was not detected in the intracuff region. Furthermore, the binding of Griffolia Simplicifolia Lectin-I (GSL-I) and endothelial nitric oxide synthase (eNOS) expression indicating the endothelial integrity was not detected in the intracuff region. Intermittent injections of ancrod, which decreases the plasma fibrinogen, inhibited occlusive thrombus formation in the intracuff region. The expression of eNOS was detected at the distal and proximal but not the intracuff region of the carotid artery treated with ancrod. Daily administration of aspirin significantly suppressed the thrombus formation in this model. These results indicate that occlusive thrombus formation accompanied by endothelial damage or dysfunction is induced by the combined application of ligation and cuff placement at the carotid artery, and suggest that this endothelial damage or dysfunction may be one pathogenesis of thrombogenesis in this model. PMID- 15334093 TI - A preemptive strike against stroke. PMID- 15334094 TI - New gene therapy technique shows some muscle. PMID- 15334095 TI - What are little boys made of? PMID- 15334096 TI - New vaccine approach targets melanoma. PMID- 15334098 TI - USDA releases animal disease risk analysis information, finalizes licensing, inspections, and procurement rules. PMID- 15334100 TI - Regulation updates. PMID- 15334101 TI - Euthanasia by cervical dislocation: when is it justified? PMID- 15334102 TI - Euthanasia by cervical dislocation: when is it justified? Where's the justification? PMID- 15334103 TI - Euthanasia by cervical dislocation: when is it justified? Submit the references. PMID- 15334104 TI - Euthanasia by cervical dislocation: when is it justified? Avoid a double standard. PMID- 15334105 TI - An external communicating lesion to the coelemic cavity in a green anole (Anolis carolensis). PMID- 15334107 TI - The top ten tips for completing the USDA annual report. PMID- 15334108 TI - Laboratory rodent transportation supplies. PMID- 15334109 TI - Discrimination of heterozygous from wild-type mice bearing a mutation in the macrophage-colony-stimulating factor gene using capillary electrophoresis. PMID- 15334110 TI - Maintaining aged rodents for biogerontology research. PMID- 15334111 TI - HSV1716 injection into the brain adjacent to tumour following surgical resection of high-grade glioma: safety data and long-term survival. AB - Following standard treatment, the prognosis remains poor in patients with high grade glioma and new therapies are urgently required. Herpes simplex virus 1716 (HSV1716) is an ICP34.5 null mutant that is selectively replication competent and shown to be safe and to replicate following injection into high-grade glioma. We demonstrate that following surgical resection, HSV1716 is safe when injected into the brain adjacent to excised tumour. In all, 12 patients with recurrent or newly diagnosed high-grade glioma underwent maximal resection of the tumour. HSV1716 was injected into eight to 10 sites around the resulting tumour cavity with the intent of infecting residual tumour cells. As clinically indicated, patients proceeded to further radiotherapy or chemotherapy. There has been no clinical evidence of toxicity associated with the administration of HSV1716. Longitudinal follow-up has allowed the assessment of overall survival compared to that of similar patients not treated with HSV1716. Three patients remain alive and clinically stable at 15, 18 and 22 months postsurgery and HSV1716 injection. Remarkably, the first patient in the trial, who had extensive recurrent disease preprocedure, is alive at 22 months since injection of HSV1716 and 29 months since first diagnosis. Imaging has demonstrated a reduction of residual tumour over the 22-month period despite no further medical intervention since the surgery and HSV1716 injection. In this study, we demonstrate that on the basis of clinical observations, there has been no toxicity following the administration of HSV1716 into the resection cavity rim in patients with high-grade glioma. The survival and imaging data, in addition to the lack of toxicity, give us confidence to proceed to a clinical trial to demonstrate efficacy of HSV1716 in glioma patients. PMID- 15334112 TI - Robust functional gene validation by adenoviral vectors: one-step Escherichia coli-derived recombinant adenoviral genome construction. AB - We describe here a clonal approach for efficient and robust construction of recombinant adenoviral genomes that holds certain advantages over existing approaches. Transgenes of interest are cloned into a small, conditionally replicating plasmid containing the left end of a recombinant adenoviral genome, encompassing pIX coding regions. Transformation of this plasmid into recombination-competent Escherichia coli bearing a plasmid containing the right end of a recombinant adenoviral genome, commencing from pIX coding regions, yields a stable co-integrated plasmid encoding a full adenoviral genome, by virtue of shared homology in pIX coding regions contained in both plasmids. The recombination process yielding the full adenoviral plasmid requires only one step, and always results in the formation of only the desired recombinant adenoviral genome. Thus, no screening is required to identify the correct plasmid encoding the desired recombinant adenoviral genome. In addition, the plasmid encoding the right-hand side of the adenoviral genome is itself incapable of producing contaminating adenovirus. We have successfully employed this approach to generate over 200 recombinant adenoviruses, obtaining only the desired recombinant adenoviral species each time. The process is amenable to medium-to high-throughput parallel construction of adenoviral genomes, and as such should aid efforts aimed towards high-throughput functional annotation of therapeutic gene targets, which aim to leverage the benefits of adenoviruses as gene delivery and expression vectors. PMID- 15334113 TI - Ultraviolet light selection assay to optimize oligonucleotide correction of mutations in endogenous xeroderma pigmentosum genes. AB - Various oligonucleotide (ODN)-based approaches have been proposed for their ability to correct mutated genes at the normal chromosomal locations. However, the reported gene correction frequencies of these approaches have varied markedly in different experimental settings, including when different tissues or cell types are targeted. In order to find the optimal ODN-based approach for a specific target tissue, an assay system that allows direct comparison of the different methods on that tissue is necessary. Herein, we describe an XP-UVC selection assay that can be used to evaluate and compare gene correction frequencies in different cell types obtained from a xeroderma pigmentosum (XP) patient, following treatment by different ODN-based approaches. As an experimental example, the XP-UVC selection assay was used to assess the ability of chimeric RNA/DNA ODN to correct point mutations in the XPA gene. This assay can be used to assess and evaluate other types of ODN-based approaches, and to further optimize them. PMID- 15334114 TI - Fc gamma R expression on NK cells influences disease severity in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is associated with autoantibodies, the best known of which is rheumatoid factor (RF). RF/IgG complexes interact with FcgammaR on the surface of neutrophils, NK cells and monocyte/macrophages. We have analyzed the expression pattern and allelic polymorphisms of three FcgammaR genes (FcgammaRIIA, FcgammaRIIC and FcgammaRIIIA) in a large sample of RA patients and normal donors. We have found that the level of FcgammaR (CD16 and CD32) expression on NK cells is lower in RA patients than in normal individuals. Genotypic analysis demonstrated that the CD32 isoform expressed by the majority of RA patients was not the activating FcgammaRIIc1 isoform, commonly seen in normal individuals, but rather the inhibitory FcgammaRIIb isoform. The combination of the FcgammaRIIIA-176F allele with a lack of CD32 expression in NK cells appeared to be characteristic of RA subjects with aggressive disease. Since FcgammaRII and FcgammaRIIIA are predominantly expressed by NK cells, these data further suggest that FcgammaR-mediated activation of NK cells could be a disease determining factor in RA patients. PMID- 15334115 TI - [Better internship--better physicians]. PMID- 15334116 TI - [Treatment of acute and chronic pain]. PMID- 15334117 TI - [Mentally ill in prisons]. PMID- 15334118 TI - [Paracetamol to preschool children--indications and cultural background]. AB - BACKGROUND: Immigrants may have a different attitude to the use of paracetamol (acetaminophen) than parents of western European or North American origin. The aim of this study was to examine under what conditions parents find treatment with paracetamol appropriate. MATERIAL AND METHOD: Parents at six public health centres were asked to select one or more among 26 indications for paracetamol treatment and to state the body temperature at which fever medication should be used. The indications were independently evaluated as appropriate, uncertain or inappropriate by the staff of the public health centres. RESULTS: 1563 (99%) parents participated in the study. 79% chose a fever threshold of 39 degrees C or above as appropriate for treatment with paracetamol. Fever, earache, influenza and headache were frequently chosen as appropriate indications by both parents and staff. On average, parents from non-western countries more frequently chose indications considered by the staff as uncertain or inappropriate and seemed to use paracetamol more frequently (odds ratio 2.35; 95% CI 1.26-4.40) and in larger doses than other parents did. INTERPRETATION: This study indicates that parents from non-western countries use paracetamol for their children on wider indications, to more of their children, and in larger doses than other parents do. PMID- 15334119 TI - [Problem forte--is paracetamol-codeine combination rational?]. AB - BACKGROUND: Combination drugs containing codeine and paracetamol are widely prescribed in Norway. MATERIAL AND METHODS: We reviewed relevant literature identified through searches on Medline or found in the reference lists of important articles. RESULTS: Codeine mediates its analgesic effect through the active metabolite morphine, a reaction which is catalysed by the cytochrome P450 isoenzyme CYP2D6. Approximately 7-10 % of the population does not express functional CYP2D6; for them codeine have no analgesic effect. They may, however, experience side effects from codeine. Used alone, codeine is an inefficient analgesic. Meta-analyses have shown little therapeutic advantage by adding codeine to paracetamol. INTERPRETATION: Codeine is an opiate with uncertain and unpredictable effects. The therapeutic benefit from the codeine component in combination with paracetamol is small, even in single dose evaluations. In chronic use, the therapeutic efficacy is most likely outweighed by side effects, including development of tolerance and abuse. PMID- 15334120 TI - [Dissemination of research--a neglected obligation]. AB - In contrast to research and teaching duties, the obligation to disseminate research results to the general public is poorly described and not lived up to in academic medicine. At the Norwegian University of Science and Technology's Faculty of Medicine, an interdisciplinary group met with academics to discuss and identify means for improvements. The academics reported positive attitudes to popularising and communicating their findings. Dissemination of research as a working area was, however, characterised as fragmented, incidental and too much dependent on individual enthusiasm. The most important initiatives in order to achieve better dissemination were said to be the following: more and better training in popularization and dealing with the media; development of an infrastructure for dissemination at a decentralised institutional level (defined responsibility, routines etc.); and encouragement of good presentations by making dissemination of research meritorious. PMID- 15334121 TI - [Mental diseases and disorders among inmates in Norwegian prisons]. AB - BACKGROUND: Surveys of serious mental disorders in prisons in the western world show high prevalences; on average 4% with psychosis, 10% with major depression, and 65% with personality disorders among male inmates. A few investigations in Norwegian prisons indicate a similar tendency. MATERIAL AND METHODS: With participation from prisons with 2850 inmates of a national total of 3000 (95%), we recorded prevalence and severity of psychiatric illnesses, other behavioural disorders, and treatment facilities available during the term served. We asked four vocational groups in each prison to reply, independently of each other, to ten semi-structured questions about these items. The groups were: prison management, guards, affiliated primary health services, and affiliated psychiatric services. RESULTS: Guards reported the highest prevalences, followed by the primary health services, then prison management; the lowest prevalence was reported by the psychiatric services. 23.5% of inmates received some kind of treatment for psychiatric disorders. INTERPRETATION: On a national scale, the average prevalence estimates for psychosis and major depression seem somewhat lower than in international surveys. Reasons for the differences in reported estimates among the vocational groups are discussed. The groups seem to have given pragmatic numbers for what they consider to be "first needs". PMID- 15334122 TI - [Are psychiatric disorders identified and treated by in-prison health services?]. AB - BACKGROUND: The prevalence in Norwegian prisons of psychiatric disorders in relation to the treatment potential in the prison health system has not been properly examined. MATERIAL AND METHOD: The prevalence of psychiatric disorders, drug problems and personality disorders was examined in a prison population in the western health region in Norway. Additionally, treatments of these disorders were surveyed. The methods used were structured clinical interviews, self reports and reviews of medical case notes. RESULTS: Psychiatric disorders in need of treatment were found in 18 out of 40 interviewed inmates. Of these 18, 13 actually received treatment with psychoactive medication. Criteria for alcohol and drug addiction or misuse were fulfilled by over 90%. Personality disorders were found in 80% and antisocial personality disorder in more than 60%. INTERPRETATION: The prevalence of psychiatric disorders including personality disorders and drug addiction is high among inmates. Compared to international studies, more of the inmates with psychiatric disorders that we interviewed receive psychoactive medication. PMID- 15334123 TI - [Nonopioid analgesics in acute pain: What has the best effect?]. PMID- 15334124 TI - [Has the Oslo96 reform brought changes in student satisfaction and study behaviour?]. AB - BACKGROUND: In 1996 the medical school of the University of Oslo introduced a new pedagogical model. The new scheme was more interactive, and partly based on a problem-based learning approach. METHOD: We used a questionnaire survey which is part of a broader study of how Norwegian professional education prepares for work. A class of medical students studying under the new scheme is compared to a class studying under the old model, as well as a class from another Norwegian medical school, that of the University of Bergen. Median values on seven-point scales are used in the comparison, as well as factor analysis with variance analysis of the factor scores. RESULTS: The overall response rate was 76%. Students studying under the new Oslo approach were more satisfied and reported more active learning strategies. They were less curriculum-oriented and used the library more often. INTERPRETATION: Some of the goals of the Oslo reform seem to have been reached. It remains to be seen whether this effect will last. PMID- 15334125 TI - [Teaching medical ethics: the Trondheim model]. AB - Medical ethics should be a core component of medical education. This paper presents a new model for teaching medical ethics in the medical school in Trondheim, Norway. The hallmark of this particular model is a systematic integration of medical ethics in the clinical arena; at the same time the theoretical foundation of medical ethics is emphasised. PMID- 15334126 TI - [Medical education in Tromso--does it secure recruitment of physicians to the northern parts of Norway?]. AB - BACKGROUND: Medical education was established in the University of Tromso in order to recruit doctors to the northern parts of Norway. Earlier publications have shown that this goal has been reached. We wanted to see whether this has remained unchanged during the last few years (1996-2001). METHOD: Our material is based on information about 318 doctors educated in Tromso and 851 doctors educated in Oslo between 1996 and 2001. The medical faculties in Tromso and Oslo provided us with doctors' names and dates of birth. The Norwegian Medical Association assisted with information about each doctor' practice and place of work. RESULTS: Among doctors educated in Tromso, the numbers going into general practice are decreasing; the majority of graduates prefer hospital work. A decreasing number take up academic work. Most of the students coming from the northern parts of Norway stay in that region upon graduation (75.4%), but the proportion is lower than earlier. Only 7.5% of doctors educated in Oslo choose to work in the northern parts of Norway. INTERPRETATION: The University of Tromso is still important in recruiting doctors to the northern regions of Norway, primarily because this university educates students who come from these regions. PMID- 15334127 TI - [50th anniversary--cheers to the internship?]. AB - This year the mandatory internship scheme for Norwegian doctors celebrates its 50th anniversary. Internships were instituted by a Decree in Council in 1954 and implemented the year after. History shows that there has been and still is a lack of commitment to the organisation as well as the content of internships. It also shows that there has been a lack of adequately formulated objectives and how detrimental this state of affairs is to the educational quality of internships. Today we face threats that the scheme may be abolished altogether. Among the reasons for this is a lack of commitment among senior doctors to supervision and mentoring, inadequate funding and, probably the most important single reason, a lack of thorough evaluation of the existent scheme. New models for internship content have been proposed by the medical authorities, but in the view of the Norwegian Medical Association the existent model should be evaluated before new models are tried out. This article shows what an important asset the internship scheme is and the necessity of arguing for keeping it. PMID- 15334128 TI - [Simulation--an efficient way of learning in graduate and continuous medical education]. AB - BACKGROUND: Different types of simulators and simulated situations are being used in the training of doctors. Even though simulation as a learning method has been used in various areas such as aviation and oil industry, it has only recently been applied to medical education. Simulation seems to be very well suited in training for and building skills and experience with life threatening situation that are seldom seen. It represents no risk to the patient and it opens up for training in realistic scenarios. METHODS: Simulations are illustrated by using Miller's learning pyramid and Bloom's cognitive levels to show how different types of simulation can be used depending on the skills that are to be acquired. Team simulation is seen as a main factor for quality improvement in medicine. INTERPRETATION: Simulation and team simulation in particular is highly rated by participating professionals. There seems to be a problem in finding objective criteria for assessing the simulated scenarios and then to generalise the findings to real situations. PMID- 15334129 TI - [Development of postgraduate medical education in Norway]. AB - Guidelines for specialist accreditation in Norway were adopted in 1918. Since then, several reviews have been made of post graduate medical education, by the Norwegian Medical Association and in cooperation with universities and public authorities. The Ministry of Social Affairs assumed responsibility for post graduate medical education in 1982. In 1998, a national council was set up as an advisory body to the Ministry of Health and tasked with monitoring specialist training and the geographical coverage of medical manpower. During the last few years, there have been proposals for extensive reforms involving the residents as well as the hospitals. This article gives a brief outline of this development. PMID- 15334130 TI - [Pathological overeating--an overview]. AB - An eating disorder apart from anorexia nervosa and bulimia nervosa is "binge eating disorder" (BED): eating in a short period of time a large quantity of food and a feeling of lack of control over food intake. There is also an atypical rest category, "eating disorders not otherwise specified" (EDNOS). Diagnostic criteria for BED and EDNOS are incomplete, particularly with respect to the definition of "bingeing" relative to bulimia nervosa. More restrictive criteria for anorexia nervosa and bulimia nervosa skew the diagnostic distribution towards BED and EDNOS, though the total prevalence of eating disorders remains unchanged. For BED and EDNOS taken together the lifetime prevalence in women is about 6%. The relationship between BED, EDNOS and overweight has mainly been overlooked; further investigations are needed. Lasting treatment effects have been found for overweight people with BED. Other eating disorders apart from anorexia nervosa and bulimia are prevalent and clinically important, and research has opened up a potential for effective treatment. PMID- 15334131 TI - [Somatic investigation and treatment of eating disorders]. AB - Eating disorders are associated with several medical complications. Growth retardation and osteoporosis can cause permanent sequelae if treatment is delayed. Severe eating disorders are associated with significant mortality. Cardiac arrhythmias are the most common somatic cause of death. Hypokalaemia is a common complication and is associated with increased risk of cardiac arrhythmias. Occasionally, overzealous refeeding may induce a potentially life-threatening condition, the refeeding syndrome. In any patient with severe eating disorder, a physician should perform diagnostic evaluation including assessment of possible somatic complications. This is necessary in order to determine where and how the patient should be treated. Most of the somatic complications of eating disorders are partly or completely reversible if the patient receives adequate treatment in time. PMID- 15334132 TI - [Eating disorders among athletes]. AB - Over the past 20 years, a number of studies have been published that generally suggest a higher frequency of eating disorders among athletes than among non athletes. Participation in competitive sport has also been considered an important factor related to the development of eating disorders. Taken together, most studies have suggested that eating disorders are particularly prevalent in sports that emphasise leanness or low body weight. However, some studies suggest a similar or lower prevalence of eating disorders compared with controls or athletes at a lower competitive level. Athletes constitute a unique population and the impact of factors such as training, eating pattern, extreme diets, restriction of food intake and psychopathological profile among them must be evaluated differently from that among non-athletes. A concerted effort by coaches, athletic trainers, parents, athletes and healthcare personnel is optimal in order to recognise, prevent and treat eating disorders in athletes. PMID- 15334133 TI - [Hospital admissions for burns]. AB - BACKGROUND: We wanted to investigate the incidence of burns and the volume of in hospital burn treatment in Norway. MATERIAL AND METHODS: Data for 1999 were compiled from hospital admissions as reported to the Norwegian Patient Register. Selection was based on ICD-10 codes for burns, though caustic injuries, reconstructive procedures, and patients discharged alive with length-of-stay less than 1 day were not included. RESULTS: 707 admissions requiring 9444 days in hospital were identified. The incidence of burns admitted to hospital was 13.5/100,000 inhabitants/year. Additionally, 102 admissions were coded as post burn reconstructive cases. 50% all admissions and 40 % of all days in hospital for burns were in hospitals without a department of plastic surgery. 24 burn patients died before discharge; 50% of those who died were above 80 years of age. 29% of all stays included a code representing surgical procedures involving skin excision and grafting. INTERPRETATION: The incidence of burns admitted to hospitals in 1999 was reduced by 20% compared to a 1977 survey. The number of reconstructive procedures was low; these options should probably be offered to more patients. We suggest that early transfer to a specialised burn centre should be considered for a somewhat larger proportion of patients. PMID- 15334134 TI - [Regulation of medical research--why and how]. PMID- 15334135 TI - [Time to disclaim responsibility a little? Physician's role as a historical construction]. PMID- 15334138 TI - [Consent and medical research--need for more nuanced rules?]. PMID- 15334141 TI - [To explain and to understand psychoses]. PMID- 15334142 TI - [Hasty conclusions on cardiac mortality in Norway]. PMID- 15334146 TI - [Empiric sepsis treatment in Norway]. PMID- 15334147 TI - [Sleepiness and fatal traffic accidents]. PMID- 15334148 TI - [Survival after high-dose therapy with autologous stem cell support--since 1981 in Norway]. PMID- 15334149 TI - [Local steroid injections in shoulder problems?]. PMID- 15334150 TI - [Ziprasidone and dementia]. PMID- 15334153 TI - Echocardiographic predictors of embolic events in infective endocarditis. AB - BACKGROUND: Systemic embolisation occurs in 22% to 50% of patients with infective endocarditis (IE). Up to 65% of embolic events (EE) involve the central nervous system which increases the mortality rate. Several echocardiographic studies have demonstrated higher embolic rates with the increase of vegetation (VEG) dimensions and mobility. AIM: To define echocardiographic parameters which can help in identifying patients with a high risk of EE and to assess the value of transesophageal echocardiography (TEE) in predicting EE in patients with IE. METHODS: 236 patients (58% male, mean age 47.8+/-6) diagnosed with IE according to Duke criteria were followed for 3 years or until cardiac surgery. Echocardiographic parameters measured on VEG included the maximum length, thickness, the narrowest diameter, neck and mobility. RESULTS: The rate of EE was 51.27% without any significant differences with respect to gender, age, fever, anaemia, VEG site or the presence of a significant regurgitation murmur. The univariate analysis showed a significant correlation between EE and IE caused by staphylococcus, IE of the right heart, and the length as well as mobility of VEG. The only independent predictors of EE were the maximum VEG length >15 mm and the increased mobility of VEG with a maximal displacement angle >60.7 degrees. In 23% of patients EE occurred after the initiation of antibiotic treatment. VEG in this group were big and very mobile (length >15 mm, maximal angle of displacement >65 degrees). CONCLUSIONS: 1. Vegetation dimension and mobility determined by TEE are important predictors of the embolic risk. 2. Significant echocardiographic predictors of embolic events included vegetation length >15 mm, neck/thickness ratio >0.69, and maximal angle of displacement of vegetation during cardiac cycle >60.7 degrees. 3. During antibiotic treatment, the embolic risk depends only on vegetation mobility and dimension. PMID- 15334154 TI - Normalisation of elevated ST segment predicts return of left ventricular systolic function and improved outcome in patients with acute myocardial infarction, treated with primary coronary angioplasty. AB - BACKGROUND: It has been shown that normalisation of elevated ST segment after primary percutaneous coronary interventions (PCI) is associated with the achievement of reperfusion at the tissue level. AIM: To assess prospectively the return of left ventricular (LV) systolic function and the outcome of patients with acute myocardial infarction (AMI), treated with primary PCI, in relation to the early normalisation of the ST segment. METHODS: The study group consisted of 110 consecutive patients (33 females, 77 males, mean age 56 years) with AMI who were successfully treated with primary PCI (TIMI flow grade 3 and residual stenosis <30%) within 12 hours from the onset of symptoms. The patients were divided into two groups according to normalisation or lack of normalisation of an elevated ST segment. The mean time from the onset of symptoms to the restoration of blood flow in an infarct-related artery (pain-to-balloon time) was similar in both groups. LV echocardiographic parameters (ejection fraction EF and wall motion score index WMSI) and clinical status directly after PCI as well as 3 and 6 months later were assessed. RESULTS: Directly after primary PCI, LV impairment (low EF and high WMSI) was significantly more often present in patients without rather than with ST segment normalisation. Afterwards, LVEF and WMSI improved significantly only in patients with ST segment normalisation whereas in the remaining patients no such improvement was observed. During 6-month follow-up period, major cardiac events (death, new AMI or need for revascularisation) occurred more frequently in patients without rather than with ST segment normalisation (p=0.03). CONCLUSIONS: 1. Rapid resolution of ST segment elevation following effective primary PCI identifies patients with a favourable outcome. 2. ST segment normalisation predicts a return of LV systolic function in patients with AMI treated with primary PCI. PMID- 15334155 TI - Relationship between alcohol consumption and cardiovascular mortality--the Warsaw Pol-MONICA Project. AB - BACKGROUND: Cardioprotective effects of alcohol recently gained wide spread interest and have been examined in several studies. AIM: To assess the effects of alcohol consumption on mortality due to cardiovascular diseases (CV) in the population of the Eastern part of Warsaw. METHODS: The study group consisted of representative, independent and randomly selected samples of the populations of two Warsaw districts (Praga Polnoc and Praga Poludnie), aged between 35 and 64 years. The studied subjects were examined in 1984 (2570 subjects), in 1988 (1397 subjects) and in 1993 (1485 subjects). Their survival rates were followed up until 1998. The annual beer, wine and vodka intake was assessed using a standardised questionnaire and calculated for a daily pure ethanol intake. The studied subjects were divided into four groups: abstinents and three groups according to the tertile distribution of the alcohol intake (mean alcohol intake in the first tertile: males 1.1 g/day, females 0.2 g/day, in the second tertile: 3.9 and 0.4 g/day, respectively, and in the third tertile: 28.2 and 2.8 g/day, respectively). The relative risk of death in the analysed groups was assessed using the proportional hazard Cox analysis. RESULTS: In total, 471 males and 244 females died during the follow-up period. There were 221 CV deaths among males and 85 among females. The relative risk of CV death after adjustment for other parameters (age, screening, cigarette smoking, body mass index, education level, cholesterol level, anginal symptoms, systolic blood pressure and self-assessed health status) was approximately 40% lower among males who consumed alcohol compared with the abstinents. The lowest risk of CV death was noted in the first tertile group. Females who consumed alcohol, had a 40-70% lower CV risk of death than abstinents the lowest risk was documented for the third tertile group. CONCLUSIONS: Alcohol consumption independently lowers the risk of death due to cardio-vascular diseases. PMID- 15334156 TI - Atherosclerotic involvement in patients with left or right dominant coronary circulation. AB - BACKGROUND: There are variations in the anatomy of the coronary arteries in patients with left dominant circulation. The influence of anatomical variations in patients with left dominant circulation on coronary arteriosclerosis is not clear. AIM: We investigated whether atherosclerotic involvement in patients with left dominant circulation differs from that in patients with right dominant circulation. METHODS: We retrospectively compared 38 consecutive angiograms with left coronary dominance with 459 consecutive angiograms with right coronary dominance. By using the 29-segment coding system of the American College of Cardiology/American Heart Association, numerical values were given to each segment, according to the percentage of the decrease in luminary diameter. Using, the sum of numerical values in each coronary artery, LAD score, LCx score, RCA score, and left main coronary artery (LMCA) score were obtained. Using the sum of these 4 vessel scores, the coronary artery disease (CAD) score was calculated. RESULTS: Total LAD score (49+/-48 vs 47+/-57), total LCx score (29+/-45 vs 41+/ 58), total RCA score (32+/-68 vs 30+/-51), total LMCA score (1.8+/-11.3 vs 2.9+/ 13.6) and total CAD score (108+/-108 vs 108+/-123) were similar in patients with left and right dominant circulation (all differences NS). CONCLUSIONS: The extent of coronary atherosclerosis does not depend on the type of dominant coronary circulation. PMID- 15334157 TI - Comparison of combined statin-fibrate treatment to monotherapy in patients with mixed hyperlipidemia. AB - BACKGROUND: Statins are the preferred drugs for the treatment of hypercholesterolemia, and fibrates for hypertriglyceridemia. In patients with mixed hyperlipidemia, monotherapy with one of these agents may not be effective and combined treatment may be preferable. AIM: To compare retrospectively the efficacy and safety of combined statin-fibrate treatment in patients with mixed hyperlipidemia in whom previous monotherapy with one of these agents occurred ineffective. METHODS AND RESULTS: The initial study group consisted of 327 patients who received micronised fenofibrate and 93 patients who received simvastatin for 12 months. Both agents caused significant changes in lipid profile. Following fibrate therapy, total cholesterol (TC), LDL-cholesterol (LDL C) and triglyceride (TG) levels decreased by 27.9%, 28.2% and 58%, respectively, and following simvastatin therapy by 33.6%, 42.8% and 37.5%, respectively. The HDL-cholesterol (HDL-C) level increased after fenofibrate by 14.8% and remained unchanged following simvastatin therapy. The TC/HDL-C ratio decreased following fenofibrate by 35.6%, and following simvastatin by 35.3%. In some patients the required reduction in lipid parameters was not achieved fenofibrate or simvastatin. Subsequently, 93 patients underwent combined therapy by adding a second agent (simvastatin in a dose of 20 mg/day or fenofibrate in a dose of 200 mg per day) which was continued for another 12 months. The addition of simvastatin to fenofibrate decreased TC, LDL-C and TG levels by 35.5%, 42.1% and 59.6%, respectively in comparison to before treatment volumes. HDL-C level was increased by 11.1%, and TC/HDL-C ratio decreased by 45.3%. The addition of fenofibrate to simvastatin decreased TC, LDL-C and TG levels by 39.3%, 48.9% and 51,6%, respectively. HDL-C level was increased by 13.4%, and TC/HDL-C ratio decreased by 49.3%. No clinical side effects nor an increase in the transaminase levels, requiring termination of the treatment, were observed. CONCLUSIONS: Combined therapy with 20 mg of simvastatin and 200 mg of micronised fenofibrate is highly effective and safe in patients with mixed hyperlipidemia. PMID- 15334158 TI - A new-onset atrial fibrillation: the incidence of potassium and magnesium deficiency. The efficacy of intravenous potassium/magnesium supplementation in cardioversion to sinus rhythm. PMID- 15334159 TI - [Aortic coarctation treated with bypass graft in adolescence and implantation of intravascular stent 12 years later--a case report]. AB - A case of a 26-year-old female with aortic coarctation and Recklinghausen disease is presented. At the age of 14 years the patient underwent thoracic aortic bypass grafting. Twelve years later she developed arterial hypertension. Spiral computerised tomography revealed patent graft and invasive hemodynamical measurements showed a gradient of 30 mmHg. The patient underwent successful intravascular stent implantation. PMID- 15334160 TI - [Polymorphic ventricular tachycardia with a short coupling interval in a patient with normal heart--a case report]. AB - We describe a case of a 59-year-old female with paroxysmal atrial fibrillation and arterial hypertension who had syncopal attacks due to polymorphic ventricular tachycardia (PMVT) with a short coupling interval of an initiating beat (280 msec). We excluded structural heart disease. In the resting ECG the QTc interval was 420 msec. During Holter monitoring a slight changes of the ST-T segment in V1 were observed (from positive T wave with ST elevation of 1 mm to flat or negative T wave without ST elevation). Additionally, after PMVT a large U-wave (4 mm of amplitude) with the QTU interval of 600 msec and QTUc interval of 662 msec were observed. The U wave disappeared 9 minutes afterwards. The ajmaline test was positive for the Brugada syndrome. The patient received ICD and sotalol, and during 6-month follow-up she remains asymptomatic. PMID- 15334161 TI - [Partial anomalous pulmonary venous drainage with atrial septal defect of the sinus venosus type. Diagnostic role of multi-slice spiral computerised tomography -a case report]. AB - A case of a 53-year-old male with a long-lasting pulmonary hypertension of an unknown cause is presented. Both transthoracic and transesophageal echocardiography as well as cardiac catheterisation failed to document any cardiac defect. Finally, a multi-slice spiral computerised tomography revealed the presence of an anomalous pulmonary vein drainage to the superior vena cava with concomitant inter-atrial septal defect of a sinus venosus type. The patient was selected for surgery. PMID- 15334162 TI - [Are "chest pain units" essential in health care systems?--American experiences, implications for Poland]. PMID- 15334163 TI - ["Chest pain units" in Polish health care systems?--Does theory translate into practice?]. PMID- 15334165 TI - [Electrocardiogram of the month]. PMID- 15334166 TI - [Infective endocarditis of the prosthetic valve]. PMID- 15334167 TI - [Angiogram of the month; unstable angina]. PMID- 15334168 TI - [An electric storm in a patient with old myocardial infarction--role of Purkinje fibers, ICD and ablation]. PMID- 15334169 TI - [Radionuclide imaging in the diagnosis of coronary artery disease in Poland in 2004]. PMID- 15334171 TI - Antiinflammatory properties of PPARgamma agonists following ischemia. AB - Ischemic disease is a leading cause of death and disability worldwide, and its incidence is expected to increase as the population ages. One population at particularly high risk of developing ischemia is patients with diabetes. Type 2 diabetes is associated with a marked increase in atherosclerosis, stroke and heart attack. Furthermore, the outcome following stroke and heart attack in diabetics is worse than in nondiabetic patients. In recent years, peroxisome proliferator-activated receptor (PPAR) agonists have been found to have potent antiinflammatory actions and have emerged as potential therapies for atherosclerosis and ischemia. The use of these agents is particularly attractive, since two PPARgamma agonists, pioglitazone (Actos) and rosiglitazone (Avandia), are already used chronically to treat diabetes. In this article we review the role of inflammation in ischemic disease and the biology of PPARs, and summarize the evidence that PPARgamma ligands suppress inflammation with an emphasis on atherosclerosis, and cerebral and myocardial ischemia. PMID- 15334172 TI - The role of the mitogen-activated protein kinase cellular signaling pathway in tumor cell survival and apoptosis. AB - A number of signaling pathways are important in the initiation and maintenance of tumor cell survival and growth. Mitogen activated protein kinases are a family of kinases of different lineages that are thought to be important in tumor growth and metastasis. These pathways provide insight into how tumor cells may escape killing by different agents. In addition, since these pathways are involved in tumor cell survival, several strategies to inhibit them are being tested for novel cancer therapies. This review summarizes the evidence for the role of these pathways in cancer and the current status of therapies. PMID- 15334173 TI - Alternative treatments for disseminated intravascular coagulation. AB - The acronym DIC is commonly interpreted as "death is coming". This pessimistic view emphasizes the deficiency of available treatment options following diagnosis of disseminated intravascular coagulation. Clinically, DIC manifests as a systemic hemorrhagic disorder associated with widespread activation and eventual exhaustion of the coagulation system, although events underlying DIC also involve effectors of inflammation. DIC can be associated with diverse conditions including sepsis and major trauma and, when identified, signifies a significant worsening in prognosis and expected mortality. Although recent clinical studies have shown that activated protein C reduces mortality in patients with severe sepsis, there is a need for further investigation and a better understanding of the underlying mechanisms. PMID- 15334174 TI - mGlu5 receptor antagonists: a novel class of anxiolytics? AB - In the early 1990s, a new family of receptors were cloned that were found to mediate the intracellular metabolic effects of glutamate via coupling to secondary messenger systems, that is, the metabotropic glutamate (mGlu) receptors. Eight such receptors (mGlu1 to mGlu8) have been cloned to date, and according to their amino acid sequence, pharmacology and second-messenger coupling, these receptors have been clustered into three groups (I-III). In contrast to the glutamate-gated ion channels (NMDA, AMPA and kainate receptors), which are responsible for fast excitatory transmission, mGlu receptors have been shown to play a modulatory role in the glutamatergic synaptic transmission either by modulating the ion channel activity or by influencing neurotransmitter release. Given the fact that the mGlu receptors are G-protein- coupled, they obviously constitute a new attractive group of "drugable" targets for the treatment of various CNS disorders. The recent discovery of small molecules that selectively bind to receptors of group I (mGlu1 and mGlu5) and group II (mGlu2 and mGlu3) allowed significant advances in our understanding of the roles of these receptors in brain physiology and pathophysiology. The identification of MPEP (2-methyl-6-(phenylethynyl)-pyridine), a highly selective and brain penetrant mGlu5 receptor antagonist, allowed the exploration of the therapeutic potential of this class of compounds. Subsequent behavior studies revealed that- with the exception of benzodiazepines--mGlu5 receptor antagonists exhibit the widest and most robust anxiolytic activity in preclinical models seen to date. Upcoming clinical studies will soon indicate if the preclinical anxiolytic-like efficacy translates into anxiolytic activity in humans. PMID- 15334175 TI - Signaling cascades implicated in muscarinic regulation of proliferation of neural stem and progenitor cells. AB - Muscarinic acetylcholine receptors (mAChRs) belong to the G-protein-coupled receptor superfamily that transduces signals through multiple intracellular signaling cascades to regulate a wide variety of physiological responses. Recently, it has been discovered that subtypes of mAChRs are expressed in proliferative neuroepithelial cells of the ventricular zone and in basic fibroblast growth factor-expanded neural stem and progenitor cell cultures. Activation of the mAChRs by ACh or its analogue carbachol leads to increased DNA synthesis and neurogenesis. The mitogenic effects of muscarinic agonists are likely mediated via mAChR-activated multisignaling pathways. The exact intracellular mechanisms underlying mAChR-modulated DNA synthesis and neurogenesis are not fully understood. However, several pathways through Ras mitogen-activated protein kinase, phosphatidylinositol 3-kinase-Akt, protein kinase C, c-Src and Ca(2+) signaling have been shown to play roles in this dynamic process. These novel signaling cascades may improve our understanding of the intracellular mechanisms underlying mAChR-stimulated neural progenitor proliferation and provide insights for therapeutic drug development. PMID- 15334176 TI - Molecule of the month. PA-457. PMID- 15334177 TI - New insights into the management of bleeding disorders. AB - The Novo Nordisk European Symposium on New Insights into the Management of Bleeding Disorders was held April 23-24, 2004, in Madrid, Spain. More than 200 specialists in the management of congenital and acquired bleeding disorders attended the different sessions. During the symposium, new medical indications for the use of human recombinant activated factor VII (rFVIIa, eptacog alfa activated) were presented. Rationale for new clinical trials and updates on registry for emerging indications were also discussed. PMID- 15334178 TI - The role of adipocytokines in adipocyte-related pathological processes. AB - It is becoming clear that adipose tissue is not merely a storage for excess energy but that it secretes a number of biologically active soluble factors collectively termed adipocytokines that control glucose and fatty acid metabolism. Of these adipocytokines, adiponectin and resistin have been the objects of intensive research, as they are implicated in obesity and diabetes related diseases. In this review, we summarize recent advances in understanding the roles of adiponectin and resistin in the causation of metabolic diseases and consider the prospects for treating metabolic disorders by targeting these two adipocytokines. PMID- 15334179 TI - The induction of heat shock proteins as a potential strategy to treat neurodegenerative disorders. AB - Neurodegenerative diseases, whether acute or chronic, are a tremendous medical problem in the modern world. Therapies are rare and only applied after a vast amount of neurons are lost. Many efforts have been made to develop new strategies to treat these disorders, but so far, there has been no breakthrough. A characteristic shared by some experimental neuroprotective substances is the induction of the heat shock response, in particular the expression of the heat shock proteins Hsp70 and Hsp27. These Hsps protect cells from cell death induced by various noxious stimuli and inhibit various cellular death pathways. Gene therapy, transgenic mice and drugs inducing Hsps in the brain decrease the infarction area after ischemia and protect neurons and nonneuronal cells of the brain. Furthermore, recent data hint toward a protective role of Hsps in chronic neurological diseases. The induction of Hsps as a possible treatment for stroke, Alzheimer's disease and Huntington's disease is discussed. PMID- 15334180 TI - Contribution of pathogens in human obesity. AB - Obesity is increasing rapidly in the United States as well as in other countries. The World Health Organization considers obesity a worldwide epidemic that poses a major public health threat. In humans, obesity causes or exacerbates a number of other diseases and co-morbidities. Etiology of obesity includes genetic, metabolic, social, behavioral and cultural factors. Although obesity has multiple causes, an often overlooked possibility is that of obesity due to an infection. Over the past two decades, seven pathogens are reported to cause obesity in animals. Canine distemper virus was the first reported obesity-promoting virus. Rous-associated virus-7, an avian retrovirus, causes stunted growth, obesity and hyperlipidemia in chickens. Borna disease virus was noted to cause obesity in rats. Scrapie agents were reported to induce obesity in mice. These pathogens appear to produce obesity by damaging the central nervous system. Next, three adenoviruses were reported to promote obesity, but their mechanisms are not clear. Animals experimentally infected with SMAM-1, an avian adeno-virus, or two human adenoviruses, adenovirus type 36 (Ad-36) and Ad-37, developed adiposity. Notably, SMAM-1 and Ad-36 were associated with obesity in humans. Although more research is needed to further define the mechanisms and the role of pathogens in the etiology of obesity, they should be included in the long list of potential etiological factors for obesity. Determination of the role of pathogens in human obesity is critical for its successful treatment and prevention. PMID- 15334181 TI - CR1-based inhibitors for prevention of complement-mediated immune hemolysis. AB - Complement receptor 1 (CR1) is a single pass transmembrane glycoprotein that, through its ability to bind key components of the complement cascade, can inhibit both the classical and alternative pathways. Using several animal models, a recombinant form of CR1 has been documented to be effective in reducing tissue damage that occurs as a result of complement activation in various inflammatory conditions. This strategy is currently being explored in human clinical trials. Activation of complement cascade via the antibody-mediated classical pathway can initiate red blood cell destruction, causing transfusion reactions and hemolytic anemia. We discuss here our approach of using CR1 derivatives as therapeutic targets for prevention of complement-dependent immune hemolysis. Using a mouse model of hemolytic transfusion reaction, we have found that sCR1 treatment reduces complement activation and prolongs the survival of transfused red blood cells. Through structure-function analysis, we have identified a complement inhibitory domain located at the amino-terminal region of CR1 that mediates its antihemolytic activity in vivo. Collectively, our data highlight a potential use for CR1 to control complement-dependent immune hemolysis and identify its functional domains for the future design of CR1-based inhibitors. PMID- 15334182 TI - The inhibition of gamma-secretase as a therapeutic approach to Alzheimer's disease. AB - Alzheimer's disease is a dementing neurodegenerative disorder for which there is no effective treatment at present. Genetic and biological studies provide evidence that the production and deposition of amyloid-beta peptides (Abeta contribute to the etiology of Alzheimer's disease. gamma-Secretase is the pivotal enzyme in generating the C terminus of Abeta which determines its aggregability and speed of deposition. Drugs that regulate the production of Abeta by inhibiting gamma-secretase activity could provide an effective therapy for Alzheimer's disease, although recent studies suggest that gamma-secretase plays important roles in cellular signaling. This review focuses on studies of the gamma-secretase biology and provides the direction for developing effective and selective gamma-secretase inhibitors as drugs for the treatment of Alzheimer's disease. PMID- 15334183 TI - Molecule of the month. Torcetrapib. PMID- 15334184 TI - Chemokines and drug discovery. AB - Chemokines and Drug Discovery was a one-day meeting organized by the Society for Medicines Research, held at the Novartis Horsham Research Centre in Horsham, United Kingdom, on March 11, 2004. More than 100 scientists, mostly from industry, attended this meeting. PMID- 15334185 TI - Current treatment and new therapies for pain management. AB - The Second Joint Scientific Meeting of the American Pain Society and the Canadian Pain Society was held May 6-9, 2004, in Vancouver, B.C., Canada. Speakers included representatives from Johns Hopkins University, Stanford University School of Medicine, Endo Pharmaceuticals and Alza Corp, among others. Highlights from the meeting are discussed. PMID- 15334186 TI - Medicine of the Maya Ameridians. AB - While perhaps best known for their advanced calendars and knowledge of astronomy, the Maya also developed an advanced pharmaceutical science that has been maintained by their descendants. Because of the diverse habitats found in the region, from mountains to deep valleys, the complex ecosystem of the highland Maya is the world's third richest in numbers of vascular plant species. The rain forests of this area are a rich source of potential new drugs. According to the U.S. National Cancer Institute, more than 25% of the ingredients in today's cancer medicines were either discovered in rain forests or are analogues or derivatives of such compounds. Although much of the Mayan medical knowledge continues to be practiced by their descendants, the Mayan medical traditions are fading because of increased urbanization. Meanwhile, the source of their pharmacopeia is disappearing as the tropical rain forests are cleared for development. Consequently, measures to preserve this pharmaceutical knowledge are of great importance. PMID- 15334187 TI - Cysteine protease cathepsin S as a key step in antigen presentation. AB - Cathepsin S is a cysteine protease in the papain super-family. Studies have shown that it is highly expressed in antigen-presenting cells. Along with other lysosomal proteases, cathepsin S plays an important role in the major histocompatibility complex class II-restricted antigen presentation, especially in the degradation of the invariant chain, a chaperone peptide bound to the class II complex. Compared with other lysosomal cysteine proteases, cathepsin S has displayed some unique characteristics. As a result, cathepsin S has been implicated as a potential target in the treatment of various disorders ranging from autoimmune diseases to atherosclerosis. Furthermore, a number of small molecule cathepsin S inhibitors have demonstrated efficacy in disease-relevant models. PMID- 15334188 TI - Vascular sphingosine-1-phosphate S1P1 and S1P3 receptors. AB - The sphingolipid sphingosine-1-phosphate (S1P) acts on five subtypes of G-protein coupled receptors, termed S1P(1) (formerly endothelial differentiation gene-1 [Edg-1]), S1P(2) (Edg-5), S1P(3) (Edg-3), S1P(4) (Edg-6) and S1P(5) (Edg-8), and possibly several other "orphan" receptors, such as GPR3, GPR6 and GPR12. These receptors are coupled to different intracellular second messenger systems, including adenylate cyclase, phospholipase C, phosphatidylinositol 3 kinase/protein kinase Akt, mitogen-activated protein kinases, as well as Rho- and Ras-dependent pathways. Consistently with this receptor multiplicity and pleiotropic signaling mechanisms, S1P influences numerous cell functions. S1P(1)1, S1P(2) and S1P(3) receptors are the major S1P receptor subtypes in the cardiovascular system, where they mediate the effects of S1P released from platelets, and possibly other tissues (such as brain). Thus S1P(1) and S1P(3) receptors enhance endothelial and vascular smooth muscle cell proliferation and migration, playing a key role in developmental and pathological angiogenesis. In contrast, S1P(2) receptors inhibit migration of these cell types, probably because of their unique stimulatory effect on a GTPase-activating protein inhibiting the activity of Rac. S1P receptors can also cause relaxation and constriction of blood vessels. The former effect is mediated by pertussis toxin sensitive receptors (possibly S1P(1)) located on the endothelium and stimulating phosphatidylinositol 3-kinase/Akt/endothelial nitric oxide synthase (eNOS). The vasoconstricting effect of S1P is likely to be mediated by S1P(2) and/or S1P(3) receptors, via Rho-Rho-kinase, and is more potent in coronary and cerebral blood vessels. Finally, S1P also protects endothelial cells from apoptosis through activation of phosphatidylinositol 3-kinase/Akt/eNOS via S1P(1) and S1P(3) receptors. The variety of these effects, taken together with the existence of multiple receptor subtypes, provides an abundance of therapeutic targets that currently still await the development of selective agents. PMID- 15334189 TI - The role of the central histaminergic system on schizophrenia. AB - Schizophrenia resistant to treatments with D(2) antagonists is thought to have the changes of extradopaminergic systems. In addition, histamine has been suggested to be a neurotransmitter in the mammalian brain that regulates many brain functions. We have recently found evidence of the role of brain histamine in schizophrenia in basic and clinical studies. Methamphetamine-induced behavioral sensitization, which has been well established as the animal model of schizophrenia, induced the enhanced histamine release, and histamine agonists inhibited the development of behavioral sensitization. As another animal model of schizophrenia, phencyclidine also increased the histamine release. In clinical studies, the levels of N-tele-methylhistamine, a major brain histamine metabolite, were elevated in the cerebrospinal fluid of schizophrenics. Moreover, H(1) receptor binding sites decreased in schizophrenics. Many atypical antipsychotics also increased histamine turnovers. Therefore, the dysfunction of the histamine neuron system may participate in the extradopaminergic brain dysfunction of schizophrenia, and histamine agents may improve the refractory schizophrenia. PMID- 15334190 TI - Molecule of the month. Rimonabant hydrochloride. PMID- 15334191 TI - Recent advances in cancer research. AB - The 95th annual meeting of the American Association for Cancer Research was held on March 27-31, 2004, in Orlando, Florida, United States. All aspects of cancer research, from drug discovery, new targets, cellular and in vivo models, clinical trials, and prevention were presented and discussed. A selection from the many excellent presentations and posters are described with an emphasis on new compounds, new methods to identify cancers, and recent discoveries that may change how we treat cancer. PMID- 15334192 TI - New developments in the treatment of neurological diseases. AB - More than 8,000 researchers, clinicians and exhibitors from around the world gathered in San Francisco for the American Academy of Neurology 56th Annual Meeting, April 24 to May 1, 2004. Of the 1,300 studies at the conference, researchers presented more than 200 abstracts each on multiple sclerosis, stroke and dementia, 145 on epilepsy, 159 on Parkinson's disease, 132 on pain and about 50 each on tremor and dystonia. The use of brain imaging technology also figured strongly in the program, with 300 abstracts that mentioned magnetic resonance imaging and 50 that included positron emission tomography. Highlights included promising Parkinson's disease studies involving gene therapy and treatments using glial-cell-derived neurotrophic factor, but also new evidence of cardiac valve regurgitation associated with pergolide. Other highlights included studies on neural repair, new guidelines for the treatment of epilepsy and important studies comparing the thrombin inhibitor ximelagatran to warfarin for the prevention of stroke. PMID- 15334193 TI - Wine, alcohol and atherosclerosis: clinical evidences and mechanisms. AB - Atherosclerosis is a chronic inflammatory disease which may cause obstructions of the coronary, cerebral and peripheral arteries. It is typically multifactorial, most often dependent on risk factors such as hypercholesterolemia, diabetes, smoking, hypertension, sedentarism, and obesity. It is the single main cause of death in most developed countries due to myocardial infarction, angina, sudden death, and heart failure. Several epidemiological studies suggest that moderate alcohol intake, especially red wine, decrease cardiac mortality due to atherosclerosis. The alcohol effect is described by a J curve, suggesting that moderate drinkers may benefit while abstainers and heavy drinkers are at higher risk. Experimental studies indicate that most beneficial effects of drinking are attributable to flavonoids that are present in red wine, purple grape juice and several fruits and vegetables. The mechanisms include antiplatelet actions, increases in high-density lipoprotein, antioxidation, reduced endothelin-1 production, and increased endothelial nitric oxide synthase expression which causes augmented nitric oxide production by endothelial cells. These findings lead to the concept that moderate red wine drinking, in the absence of contraindications, may be beneficial to patients who are at risk of atherosclerotic cardiovascular events. Moreover, a diet based on fruits and vegetables containing flavonoids may be even more beneficial. PMID- 15334194 TI - Comments on the paper "High doses of riboflavin and the elimination of dietary red meat promote the recovery of some motor functions in Parkinson's disease patients. C.G. Coimbra and V.B.C. Junqueira. Brazilian Journal of Medical and Biological Research, 36: 1409-1417, 2003". PMID- 15334195 TI - Preservation of graft function in low-risk living kidney transplant recipients treated with a combination of sirolimus and cyclosporine. AB - The use of sirolimus (SRL) in combination with full doses of cyclosporin A (CsA) results in reduced one-year kidney allograft function, which is associated with shorter long-term allograft survival. We determined the effect of reduced CsA exposure on graft function in patients receiving SRL and prednisone. Ninety recipients of living kidney transplants receiving SRL (2 mg/day, po) were compared to 35 recipients receiving azathioprine (AZA, 2 mg kg-1 day-1, po). All patients also received CsA (8-10 mg kg-1 day-1, po) and prednisone (0.5 mg kg-1 day-1). Efficacy end-point was a composite of biopsy-confirmed acute rejection, graft loss, or death at one year. Graft function was measured by creatinine, creatinine clearance, and graft function deterioration between 3 and 12 months (delta1/Cr). CsA concentrations in patients receiving SRL were 26% lower. No differences in one-year composite efficacy end-point were observed comparing SRL and AZA groups (18 vs 20%) or in the incidence of biopsy-proven acute rejection (14.4 and 14.3%). There were no differences in mean +/- SD creatinine (1.65 +/- 0.46 vs 1.60 +/- 0.43 mg/dl, P = 0.48) or calculated creatinine clearances (61 +/ 15 vs 62 +/- 13 ml/min, P = 0.58) at one year. Mean +/- SD delta1/Cr (-11 +/- 17 vs -14 +/- 15%, P = 0.7) or the percentage of patients with >20% (26 vs 31%, P = 0.6) or >30% delta1/Cr (19 vs 17%, P = 1) did not differ between the two groups. The use of 2-mg fixed oral doses of SRL and reduced CsA exposure was effective in preventing acute rejection and preserving allograft function. PMID- 15334196 TI - Expression of inducible nitric oxide synthase is increased in patients with heart failure due to ischemic disease. AB - The objective of the present study was to determine the relationship between nitric oxide synthases (NOS) and heart failure in cardiac tissue from patients with and without cardiac decompensation. Right atrial tissue was excised from patients with coronary artery disease (CAD) and left ventricular ejection fraction (LVEF) <35% (N = 10), and from patients with CAD and LVEF >60% (N = 10) during cardiac surgery. NOS activity was measured by the conversion of L-[H(3)] arginine to L-[H(3)]-citrulline. Gene expression was quantified by the competitive reverse transcription-polymerase chain reaction. Both endothelial NOS (eNOS) activity and expression were significantly reduced in failing hearts compared to non-failing hearts: 0.36 +/- 0.18 vs 1.51 +/- 0.31 pmol mg-1 min-1 (P < 0.0001) and 0.37 +/- 0.08 vs 0.78 +/- 0.09 relative cDNA absorbance at 320 nm (P < 0.0001), respectively. In contrast, inducible NOS (iNOS) activity and expression were significantly higher in failing hearts than in non-failing hearts: 4.00 +/- 0.90 vs 1.54 +/- 0.65 pmol mg-1 min-1 (P < 0.0001) and 2.19 +/- 0.27 vs 1.43 +/- 0.13 cDNA absorbance at 320 nm (P < 0.0001), respectively. We conclude that heart failure down-regulates both eNOS activity and expression in cardiac tissue from patients with LVEF <35%. In contrast, iNOS activity and expression are increased in failing hearts and may represent an alternative mechanism for nitric oxide production in heart failure due to ischemic disease. PMID- 15334197 TI - Patient and physician evaluation of the severity of acute asthma exacerbations. AB - We studied the ability of patients not experienced in the use of peak expiratory flow meters to assess the severity of their asthma exacerbations and compared it to the assessment of experienced clinicians. We also evaluated which data of physical examination and medical history are used by physicians to subjectively evaluate the severity of asthma attacks. Fifty-seven adult patients (15 men and 42 women, with a mean (+/- SD) age of 37.3 +/- 14.5 years and 24.0 +/- 17.9 years of asthma symptoms) with asthma exacerbations were evaluated in a University Hospital Emergency Department. Patients and physicians independently evaluated the severity of the asthma attack using a linear scale. Patient score, physician score and forced expiratory volume at the first second (FEV1) were correlated with history and physical examination variables, and were also considered as dependent variables in multiple linear regression models. FEV1 correlated significantly with the physician score (rho = 0.42, P = 0.001), but not with patient score (rho = 0.03; P = 0.77). Use of neck accessory muscles, expiratory time and wheezing intensity were the explanatory variables in the FEV1 regression model and were also present in the physician score model. We conclude that physicians evaluate asthma exacerbation severity better than patients and that physician's scoring of asthma severity correlated significantly with objective measures of airway obstruction (FEV1). Some variables (the use of neck accessory muscles, expiratory time and wheezing intensity) persisted as explanatory variables in physician score and FEV1 regression models, and should be emphasized in medical schools and emergency settings. PMID- 15334198 TI - Factors associated with emergency department visits due to acute asthma. AB - It is important to identify characteristics related to poor disease control and frequent visits to the emergency department (ED). The objective of the present study was to compare the characteristics of patients attending the adult ED for treatment of asthma exacerbation with those attending an asthma specialist clinic (AC) in the same hospital, and to determine the factors associated with frequent visits to the ED. We conducted a cross-sectional survey of consecutive patients (12 years and older) attending the ED (N = 86) and the AC (N = 86). Significantly more ED patients than AC patients reported ED visits in the past year (95.3 vs 48.8%; P < 0.001) and had difficulty performing work (81.4 vs 49.4%; P < 0.001. Significantly more AC than ED patents had been treated with inhaled corticosteroids (75.6 vs 18.6%; P < 0.001) used to increase or start steroid therapy when an attack was perceived (46.5 vs 20.9%; P < 0.001) and correctly used a metered-dose inhaler (50.0 vs 11.6%; P < 0.001). The history of hospital admissions (odds ratio, OR, 4.00) and use of inhaled corticosteroids (OR, 0.27) were associated with frequent visits to the ED. In conclusion, ED patients were more likely than AC patients to be dependent on the acute use of the ED, were significantly less knowledgeable about asthma management and were more likely to suffer more severe disease. ED patients should be considered an important target for asthma education. Facilitating the access to ambulatory care facilities might serve to reduce asthma morbidity. PMID- 15334199 TI - Clonal dissemination of VanA-type glycopeptide-resistant Enterococcus faecalis between hospitals of two cities located 100 km apart. AB - Nosocomial dissemination of glycopeptide-resistant enterococci represents a major problem in hospitals worldwide. In Brazil, the dissemination among hospitals in the city of Sao Paulo of polyclonal DNA profiles was previously described for vancomycin-resistant Enterococcus faecium. We describe here the dissemination of VanA phenotype E. faecalis between two hospitals located in different cities in the State of Sao Paulo. The index outbreak occurred in a tertiary care university hospital (HCUSP) in the city of Sao Paulo and three years later a cluster caused by the same strain was recognized in two patients hospitalized in a private tertiary care hospital (CMC) located 100 km away in the interior of the state. From May to July 1999, 10 strains of vancomycin-resistant E. faecalis were isolated from 10 patients hospitalized in the HCUSP. The DNA genotyping using pulsed-field gel electrophoresis (PFGE) showed that all isolates were originated from the same clone, suggesting nosocomial dissemination. From May to July 2002, three strains of vancomycin-resistant E. faecalis were isolated from two patients hospitalized in CMC and both patients were colonized by the vancomycin-resistant Enterococcus in skin lesions. All isolates from CMC and HCUSP were highly resistant to vancomycin and teicoplanin. The three strains from CMC had minimum inhibitory concentration >256 micro g/ml for vancomycin, and 64 (CMC 1 and CMC 2) and 96 micro g/ml (CMC 3) for teicoplanin, characterizing a profile of VanA resistance to glycopeptides. All strains had the presence of the transposon Tn1546 detected by PCR and were closely related when typed by PFGE. The dissemination of the E. faecalis VanA phenotype among hospitals located in different cities is of great concern because E. faecalis commonly colonizes the gastrointestinal tract of patients and healthy persons for periods varying from weeks to years, which, together with the persistence of vancomycin-resistant Enterococcus in hospital rooms after standard cleaning procedures, increases the risk of the dissemination and reservoir of the bacteria. PMID- 15334200 TI - Use of molecular epidemiology to monitor the nosocomial dissemination of methicillin-resistant Staphylococcus aureus in a university hospital from 1991 to 2001. AB - Methicillin-resistant Staphylococcus aureus (MRSA) has been the cause of major outbreaks and epidemics among hospitalized patients, with high mortality and morbidity rates. We studied the genomic diversity of MRSA strains isolated from patients with nosocomial infection in a University Hospital from 1991 to 2001. The study consisted of two periods: period I, from 1991 to 1993 and period II from 1995 to 2001. DNA was typed by pulsed-field gel electrophoresis and the similarity among the MRSA strains was determined by cluster analysis. During period I, 73 strains presented five distinctive DNA profiles: A, B, C, D, and E. Profile A was the most frequent DNA pattern and was identified in 55 (75.3%) strains; three closely related and four possibly related profiles were also identified. During period II, 80 (68.8%) of 117 strains showed the same endemic profile A identified during period I, 18 (13.7%) closely related profiles and 18 (13.7%) possibly related profiles and, only one strain presented an unrelated profile. Cluster analysis showed a 96% coefficient of similarity between profile A from period I and profile A from period II, which were considered to be from the same clone. The molecular monitoring of MRSA strains permitted the determination of the clonal dissemination and the maintenance of a dominant endemic strain during a 10-year period and the presence of closely and possibly related patterns for endemic profile A. However, further studies are necessary to improve the understanding of the dissemination of the endemic profile in this hospital. PMID- 15334201 TI - Normal expression of IFN-gammaR in four patients with uncommon mycobacterial infection phenotypes. AB - Several primary immunodeficiency diseases affecting the interleukin 12/interferon gamma (IFN-gamma) pathway have been identified, most of them characterized by recurrent and protracted infections produced by intracellular microorganisms, particularly by several species of mycobacteria. In the present study we analyzed the expression of IFN-gamma receptor (IFN-gammaR) and signal transducer and activator of transcription 1 (STAT-1) in 4 children with Mycobacterium tuberculosis infection of uncommon clinical presentation. These molecules were evaluated by flow cytometry and Western blotting in B cells transformed with Epstein-Barr virus and mutations were scanned by single-strand conformational polymorphisms and DNA sequencing. The expression of IFN-gammaR1 was normal in all 4 patients. The genetic analysis of IFN-gammaR1 and IFN-gammaR2 coding sequences did not reveal any mutation. The expression of the STAT-1 molecule was similar in patients and healthy controls; however, when the phosphorylation of this transcription factor in response to IFN-gamma activation was evaluated by Western blot, a significant lower signal was evident in one patient. These data indicate that there are no alterations in the expression or function of the IFN-gammaR chains in these patients. However, the low level of STAT-1 phosphorylation found in one of these patients might be explained by a defect in one of the molecules involved in the signal transduction pathway after IFN-gamma interacts with its receptor. In the other three patients the inability to eliminate the mycobacteria may be due to a defect in another effector mechanism of the mononuclear phagocytes. PMID- 15334202 TI - Cyclophosphamide in the treatment of focal segmental glomerulosclerosis. AB - Prednisone is the initial treatment of primary focal segmental glomerulosclerosis. However, when immunosuppressive agents in combination with steroids are used in the treatment of prednisone-dependent and prednisone resistant patients the remission rate is variable. We report a long-term trial using cyclophosphamide (2.0 to 3.0 mg/kg body weight for 12 weeks) in combination with prednisone (1.0 to 2.0 mg/kg body weight), as compared with prednisone alone for the treatment of prednisone-resistant and frequently relapsing nephrotic syndrome and focal segmental glomerulosclerosis. Fifty-four patients (34 males and 20 females) with a diagnosis of idiopathic nephrotic syndrome and focal segmental glomerulosclerosis, followed-up for an average of 86.1 +/- 82.4 months, were evaluated. Complete remission occurred in 20.4% and partial remission in 14.8% of the patients treated with steroids and in 26.7 and 20.0% of the patients treated with cyclophosphamide + prednisone, respectively. Of the 24 prednisone resistant patients treated with steroids in combination with cyclophosphamide, 33.3% obtained a complete/partial response. At the time of final evaluation, 25% of the patients treated with prednisone and 10.0% of those treated with prednisone in combination with cyclophosphamide had reached end-stage renal disease. Persistent nephrotic syndrome and progressive renal insufficiency were more frequently observed among the patients treated with prednisone alone (50.0 vs 33.3% and 33.3 vs 16.7%, respectively). The treatments were well tolerated and no patient experienced adverse reactions requiring discontinuation of medications. Although open-label and non-randomized, the present trial showed that cyclophosphamide is a reasonable choice for the treatment of primary focal segmental glomerulosclerosis and prednisone-resistant nephrotic syndrome. PMID- 15334203 TI - Comparative study of IgA nephropathy with and without crescents. AB - Glomerular crescents were analyzed as a prognostic factor in retrospectively reviewed data from 144 patients with biopsy-proven IgA nephropathy. Crescents were found in 26 (18%) patients, and detected in 2 to 100% of glomeruli in each specimen. In 5% of the patients more than 50% of the glomeruli were affected. Thirty patients with IgA nephropathy without crescents were studied as a control group. Mean age was 30.3 +/- 9.4 and 30.2 +/- 12.0 years for the patients with and without crescents, respectively, and males prevailed in both groups. The length of follow-up was 23.2 +/- 41.6 months for patients with crescents and 29.3 +/- 35.3 months for patients without crescents. Eighty percent of the patients with crescents were hypertensive, compared to 27% of the non-crescent control group (P < 0.05). Mean serum creatinine at the time of diagnosis was 3.9 +/- 2.9 and 1.9 +/- 2.1 mg/dl for the patients with and without crescents, respectively. Initial urinary protein excretion was higher in patients with crescents (4.6 +/- 3.5 vs 1.2 +/- 0.9 g/day; P < 0.05). At the end of follow-up 17 patients (77.3%) from the crescent group and 3 (11.1%) from the non-crescent group had end-stage renal disease (P < 0.0001). The presence of crescents was associated with higher levels of initial serum creatinine and urinary protein excretion, and a higher frequency of hypertension and progression to end-stage renal disease. PMID- 15334204 TI - Response to an oral calcium load in nephrolithiasis patients with fluctuating parathyroid hormone and ionized calcium levels. AB - The response to an oral calcium load test was assessed in 17 hypercalciuric nephrolithiasis patients who presented elevated parathyroid hormone (PTH) irrespective of the ionized calcium (sCa2+) levels. Blood samples were collected at baseline (0 min) and at 60 and 180 min after 1 g calcium load for serum PTH, total calcium, sCa2+, and 1.25(OH)2D3 determinations. According to the sCa2+ level at baseline, patients were classified as normocalcemic (N = 9) or hypercalcemic (N = 8). Six healthy subjects were also evaluated as controls. Bone mineral density was reduced in 14/17 patients. In the normocalcemic group, mean PTH levels at 0, 60 and 180 min (95 +/- 76, 56 +/- 40, 57 +/- 45 pg/ml, respectively) did not differ from the hypercalcemic group (130 +/- 75, 68 +/- 35, 80 +/- 33 pg/ml) but were significantly higher compared to healthy subjects despite a similar elevation in sCa2+ after 60 and 180 min vs baseline in all 3 groups. Mean total calcium and 1.25(OH)2D3 were similar in the 3 groups. Additionally, we observed that 5 of 9 normocalcemic patients presented a significantly higher concentration-time curve for serum PTH (AUC0',60',180') than the other 4 patients and the healthy subjects, suggesting a primary parathyroid dysfunction. These data suggest that the individual response to an oral calcium load test may be a valuable dynamic tool to disclose a subtle primary hyperparathyroidism in patients with high PTH and fluctuating sCa2+ levels, avoiding repeated measurements of both parameters. PMID- 15334205 TI - Contrast sensitivity threshold measured by sweep-visual evoked potential in term and preterm infants at 3 and 10 months of age. AB - Although healthy preterm infants frequently seem to be more attentive to visual stimuli and to fix on them longer than full-term infants, no difference in visual acuity has been reported compared to term infants. We evaluated the contrast sensitivity (CS) function of term (N = 5) and healthy preterm (N = 11) infants at 3 and 10 months of life using sweep-visual evoked potentials. Two spatial frequencies were studied: low (0.2 cycles per degrees, cpd) and medium (4.0 cpd). The mean contrast sensitivity (expressed in percentage of contrast) of the preterm infants at 3 months was 55.4 for the low spatial frequency (0.2 cpd) and 43.4 for the medium spatial frequency (4.0 cpd). At 10 months the low spatial CS was 52.7 and the medium spatial CS was 9.9. The results for the term infants at 3 months were 55.1 for the low spatial frequency and 34.5 for the medium spatial frequency. At 10 months the equivalent values were 54.3 and 14.4, respectively. No difference was found using the Mann-Whitney rank sum T-test between term and preterm infants for the low frequency at 3 or 10 months or for the medium spatial frequency at 3 or 10 months. The development of CS for the medium spatial frequency was equally fast for term and preterm infants. As also observed for visual acuity, CS was equivalent among term and preterm infants, suggesting that visual experience does not modify the development of the primary visual pathway. An earlier development of synapses in higher cortical visual areas of preterm infants could explain the better use of visual information observed behaviorally in these infants. PMID- 15334206 TI - Immunoexpression of CD95 in chronic gastritis and gastric mucosa-associated lymphomas. AB - CD95 (Fas/APO-1)-mediated apoptosis plays an important role in immunological regulation and is related to the pathogenesis of autoimmune diseases. Immunoexpression of CD95 has been reported to frequently occur in low grade non Hodgkin lymphomas, especially of post-germinal center histogenesis, among which those originating in mucosa-associated lymphoid tissue (MALT lymphomas). However, there is no report comparing in situ immunoexpression of this marker in lymphomas and the hyperplastic lymphoid reaction (chronic gastritis) related to Helicobacter pylori infection. The purpose of the present research was to compare the intensity of lymphoid CD95 immunoexpression in 15 cases of H. pylori-related chronic gastritis and 15 gastric MALT lymphomas. CD95 (anti-CD95) was detected by an immunoperoxidase technique in paraffin sections using the catalyzed amplification system. Graduation of reaction intensity (percentage of CD95 positive cells) was semiquantitative, from 1+ to 4+. Nine cases of chronic gastritis were 4+, five 2+ and one 1+. Three lymphomas were 4+, three 3+, four 2+, four 1+, and one was negative. Although 14 of 15 lymphomas were positive for CD95, the intensity of the reaction was significantly weaker compared to that obtained with gastric tissue for patients with gastritis (P = 0.03). The difference in CD95 immunoexpression does not seem to be useful as an isolated criterion in the differential diagnosis between chronic gastritis and MALT lymphomas since there was overlapping of immunostaining patterns. However, it suggests the possibility of a pathogenetic role of this apoptosis-regulating protein in MALT lymphomas. PMID- 15334207 TI - The relationship between salivary cortisol concentrations and anxiety in adolescent and non-adolescent pregnant women. AB - The main purpose of the present study was to determine the relationship between salivary cortisol concentrations and self-report anxiety in 50 adolescent and 178 non-adolescent women during the last month of pregnancy. The subjects were randomly selected from a previous study involving women who attended antenatal care from September 1997 to August 2000 at 17 health services in Southeast Brazil. Salivary cortisol was measured with an enzyme immunoassay kit, and anxiety was assessed by the State-Trait Anxiety Inventories (STAI) of Spielberger. After saliva collection, the participants completed the STAI. Mean concentrations of cortisol for both pregnant adolescents (14.17 +/- 6.78 nmol/l) and non-adolescents (13.81 +/- 8.51 nmol/l) were similar (P = 0.89). Forty-three percent of the pregnant adolescents and 30.5% of the non-adolescents felt anxious at the time of being questioned (State Anxiety Inventory (SAI) scores >40; P = 0.06). Cortisol concentrations in adolescents were negatively related to the SAI scores (r = -0.39; P = 0.01) which assess a temporary condition of anxiety. There was a statistically significant difference in mean cortisol concentrations between adolescents with low (<=40) and high (>40) SAI scores (P = 0.03, t-test), but no differences for non-adolescents. The negative relationship between salivary cortisol concentrations and anxiety scores in adolescents may be due to puberty-related hormone differences during this period of life. Pregnant adolescents may possess unique biological or psychological characteristics compared to adults and non-pregnant adolescents. Thus, we need to know more about the hypothalamic-pituitary-adrenocortical axis of pregnant adolescents. PMID- 15334208 TI - Brazilian version of the Berg balance scale. AB - The purpose of the present study was to translate and adapt the Berg balance scale, an instrument for functional balance assessment, to Brazilian-Portuguese and to determine the reliability of scores obtained with the Brazilian adaptation. Two persons proficient in English independently translated the original scale into Brazilian-Portuguese and a consensus version was generated. Two translators performed a back translation. Discrepancies were discussed and solved by a panel. Forty patients older than 65 years and 40 therapists were included in the cultural adaptation phase. If more than 15% of therapists or patients reported difficulty in understanding an item, that item was reformulated and reapplied. The final Brazilian version was then tested on 36 elderly patients (over age 65). The average age was 72 years. Reliability of the measure was assessed twice by one physical therapist (1-week interval between assessments) and once by one independent physical therapist. Descriptive analysis was used to characterize the patients. The intraclass correlation coefficient (ICC) and Pearson's correlation coefficient were computed to assess intra- and interobserver reliability. Six questions were modified during the translation stage and cultural adaptation phase. The ICC for intra- and interobserver reliability was 0.99 (P < 0.001) and 0.98 (P < 0.001), respectively. The Pearson correlation coefficient for intra- and interobserver reliability was 0.98 (P < 0.001) and 0.97 (P < 0.001), respectively. We conclude that the Brazilian version of the Berg balance scale is a reliable instrument to be used in balance assessment of elderly Brazilian patients. PMID- 15334209 TI - Correlation between serum E-selectin levels and panoramic nailfold capillaroscopy in systemic sclerosis. AB - E-selectin is expressed by the activated endothelium and its plasma levels are increased in patients with systemic sclerosis. Eighteen patients fulfilling the American Rheumatism Association criteria for systemic sclerosis, 15 females and 3 males, 42-70 years old, 9 with diffuse and 9 with limited forms, were sequentially recruited for this study. Serum E-selectin levels were determined by commercially available ELISA and their association with nailfold capillaroscopic abnormalities was investigated. Nailfold capillaries were analyzed by 16X magnification wide-field capillaroscopy. Two parameters on capillaroscopy were used to correlate to serum E-selectin: deletion and ectasia. Data were analyzed statistically by the Student t-test and Spearman correlation. Two-tailed P values below 0.05 were considered significant. E-selectin range was 38 to 200 ng/ml (80 +/- 39.94). There was a correlation between serum E-selectin levels and the deletion capillaroscopic score (r = 0.50, P < 0.035). This correlation was even stronger within the first 48 months of diagnosis (r = 0.63, P < 0.048). On the other hand, no association was observed between selectin and ectasia. Patients with diffuse disease presented higher serum E-selectin levels than patients with limited disease, although the difference was not statistically significant (96.44 +/- 48.04 vs 63.56 +/- 21.77 ng/dl; P = 0.08). The present study is the first showing a correlation between soluble serum E-selectin levels and alterations in capillaroscopy. The stronger correlation of deletion score in capillaroscopy in early disease suggests that serum E-selectin levels might be a useful biochemical marker of disease activity in systemic sclerosis. PMID- 15334210 TI - Near nerve potential of sural nerve in leprosy. AB - Leprosy neuropathy is characterized by initial involvement of the small nerve fibers, later followed by involvement of the large fibers, when routine nerve conduction studies become abnormal. To increase the diagnostic yield and precocity of these studies, we applied the near nerve technique to the sural nerve of 8 leprosy patients. Contrary to our expectations, the main component of the sural nerve sensory action potential was abnormal in all patients, but the minimum conduction velocity originating from small 3-6 mm fibers was normal or only mildly involved in three patients. Also, although Schwann cells are the first to be involved in leprosy, the results are suggestive of axonal degeneration instead of demyelination. To better understand the neurophysiology and physiology of leprosy and to increase the accuracy and precocity of the diagnosis, it will be necessary to investigate patients in the very early stages of the disease and to correlate these findings with the corresponding nerve pathology. PMID- 15334211 TI - Visual event-related potential (P300): a normative study. AB - The P300 component of the Event-Related Potential (ERP) is a general measurement of "cognitive efficiency". It is an index of the ability of an individual's Central Nervous System (CNS) to process incoming information. OBJECTIVE: To develop a normative database for the visual P300. METHODOLOGY: 30 right-handed individuals (same number of each sex), between 20 and 30 years of age, healthy, free of any cognitive impairment and not making use of psychoactive substances. Participants were submitted to a visual discrimination task, which employed the "oddball" paradigm. RESULTS: The expected scalp distribution trend was seen for latency but not for amplitude values. CONCLUSION: A high variability of latency and amplitude values was observed across the age span. Mean reaction time for the entire sample of the study was 391.56 +/- 37.03 ms. PMID- 15334212 TI - The Wada Test: contributions to standardization of the stimulus for language and memory assessment. AB - The Wada Test (WT) is part of the presurgical evaluation for refractory epilepsy. The WT is not standardized and the protocols differ in important ways, including stimulus type of material presented for memory testing, timing of presentations and methods of assessment. The aim of this study was to contribute to establish parameters for a WT to Brazilian population investigating the performance of 100 normal subjects, without medication. Two parallel models were used based on Montreal Procedure adapted from Gail Risse's (MEG-MN,EUA) protocol. The proportions of correct responses of normal subjects submitted to two parallel WT models were investigated and the two models were compared. The results showed that the two models are similar but significant differences among the stimulus type were observed. The results suggest that the stimulus type may influence the results of the WT and should be considered when constructing models and comparing different protocols. PMID- 15334213 TI - Headache epidemiology in Vitoria, Espirito Santo. AB - This is the first headache survey in the region of Vitoria, ES Brazil. A high prevalence of headache sufferers was found (52.8%). Headache was more common among women (63.9%) and less common among people older than 55 years old. The type of professional activity was not related with the headache prevalence. The most frequent causal attribution was stress. Most headache sufferers are not under regular medical treatment (9%), and most of them use analgesic drugs without proper orientation. The most used compounds are combinations with caffeine (33%) and simple analgesics (52.3%). PMID- 15334214 TI - Prevalence of presenile dementia in a tertiary outpatient clinic. AB - There are very few reports about prevalence of presenile dementia in Brazil. We reviewed files of patients evaluated with early onset of cognitive impairment in our institution. Among 141 patients (61% males) there was no difference between gender by age at onset or at first evaluation. We have observed an increasing number of patients after 50 years. The most frequent causes were: vascular dementia (36.9%), Alzheimer's disease (20.3%) and traumatic brain injury (9.2%). There was difference among dementia type by age of onset and first evaluation, educational level and length of dementia. These results may be compared with those from other neurologic services in order to replicate or confirm these results. PMID- 15334215 TI - Seasonal distribution and demographical characteristics of carpal tunnel syndrome in 1039 patients. AB - OBJECTIVE: To describe the demographic characteristics of gender, age, and presence of repetitive movements, intake of alcohol and non-steroid anti inflammatories (NSAI), medical specialties that referred patients to nerve conduction studies and electromyography (NCS-EMG), school attainment, and seasonal distribution in patients with a neurophysiological diagnosis of carpal tunnel syndrome (CTS) in the State of Rio Grande do Sul, Brazil. METHOD: A series of 1039 patients (1549 hands) with neurophysiologically defined CTS was studied. Patients were referred for NCS-EMG in 3 universities and 2 private services, from August 2001 to January 2003. All patients completed a questionnaire containing demographic information. The diagnosis of CTS was established following a pre established protocol, with defined diagnostic criteria. RESULTS: Around one fourth of patients had already performed NCS-EMG; the greatest frequency of CTS was observed in women (5.6:1) and in patients above the age of 40. Most patients reported performing repetitive movements in their daily routine (69.7%); 12.9% reported use of NSAI and 14.9% regular intake of alcoholic beverages. A greater frequency of CTS was observed in the months of July and August, when compared to the other months of the year. Around 2/3 of the study population had completed at least secondary school. Most requests of nerve conduction studies did not provide a diagnostic hypothesis (59.9%) and neurophysiologic studies were requested mostly by traumatology/orthopedics (71.1%). CONCLUSION: We have concluded that, in our environment, CTS shows some demographical characteristics that are similar to what the literature describes. Also, we have found that most of our sample concluded at least secondary school, and was referred to neurophysiologic studies by orthopedists. To be pointed out is the seasonal distribution of CTS, which demonstrates a significant association with winter months. PMID- 15334216 TI - Papillomas and carcinomas of the choroid plexus: histological and immunohistochemical studies and comparison with normal fetal choroid plexus. AB - BACKGROUND: Choroid plexus tumors are rare. Results on immunohistochemical features are scanty and controversial even regarding normal plexus. METHOD: Thirteen cases of choroid plexus tumors and five samples of normal fetal choroid plexus were submitted to immunohistochemical study using a panel of epithelial, neuronal and stromal markers. RESULTS/CONCLUSIONS: Relevant histological findings were presence of clear cells in 3/5 papillomas (PP) and 7/8 carcinomas (CA) and all 5 fetal plexuses; rhabdoid cells, desmoplasia and vascular proliferation were found respectively in 3, 4 and 5 cases out of 6 poorly differentiated CA and were absent in PP and well differentiated CA. Pancytokeratin AE1/AE3 was strongly positive in all 13 cases, even in the undifferentiated component of poorly differentiated CA, where reactivity was focal in 3 and diffuse in 3 cases. Low molecular weight cytokeratin (35betaH11) was not expressed in any of the 8 CA, but was present in all 5 PP. In 4 of 6 poorly differentiated CA there was reactivity for smooth muscle actin (1A4) in 10 to 30% of the cells. This was true also for one case lacking rhabdoid cells. Laminin was undetectable in all 6 cases of poorly differentiated CA but was present in 4 PP and 2 well differentiated CA. All 5 fetal plexuses expressed GFAP. PMID- 15334217 TI - Gliosarcoma: report of four cases with immunohistochemical findings. AB - Gliosarcoma (GSa) is a rare primary central nervous system neoplasm (CNS) characterized by biphasic histological pattern with both glial and sarcomatous components. Our objective is to describe the clinical, morphological and immunohistochemical features of four cases of GSa and to discuss its pathogenetic mechanisms. The male:female ratio was 3:1. The mean age was 39 years, ranging from 19 to 48. Headache was the commonest clinical symptom. All patients underwent craniotomy with microsurgery and total resection of the tumor. Diagnosis was suspected due to microscopic architecture and confirmed by detection of reticulin fibers through histochemical techniques. Immunohistochemical analysis was positive for p53 in both glial and sarcomatous cells in all four cases. EGFR was focally positive in glial cells in one case. Our findings support monoclonal origin of GSa involving the TP53 tumor-suppressor gene. However, alternative pathways cannot be ruled out. PMID- 15334218 TI - Spontaneous extracranial carotid atherosclerosis evolution in asymptomatic individuals: a three-year prospective study. AB - OBJECTIVE: To evaluate the spontaneous evolution of extracranial carotid atherosclerosis in asymptomatic patients who did not present the main risk factors associated to the disease. METHOD: A prospective study including patients of both genders, age ranging from 40 to 70 years, not presenting any signs and symptoms of cerebrovascular disease and without the main atherosclerosis risk factors were included. Patients who were using or had used medication during the follow-up period that could potentially influence in the spontaneous course of atherosclerosis were excluded. The evaluation of the plaque and degree of stenosis were acquired using mode B, 7.5 MHz Doppler ultrasonography (USG). The follow-up was carried out for 36 months, with clinical, neurological, and USG exams repeated in a period of 6 to 8 months. Ninety-six individuals (48 women) completed the study with the presence of plaque, and 52 (26 women) with a degree of stenosis. RESULTS: As to the degree of stenosis, 25% of the patients had worsening, 69% remained stable and 6% improved. When only the presence or absence of plaque was considered, 20% showed worsening (plaque developed during follow up), 7% improved (disappearance of plaque), and 73% remained stable. No differences were found between the male and female patients. CONCLUSION: These results confirm the dynamic characteristics of plaque. In asymptomatic individuals without specific treatment, spontaneous improvement may occur, however, rarely. These findings may contribute as an assessment criterion when a decision is to be made in high-risk patients. PMID- 15334219 TI - Ischemic stroke in children: a study of the associated alterations. AB - Arterial ischemic stroke (AIS) in children is a relatively rare disease, not yet clearly understood and with a multifactored etiology. It can cause a severe impact on the child and be the first manifestation of a systemic disease. Delayed diagnosis is still common and research on the subject in our field practically does not exist. Prothrombotic disorders have been described as important causative factors of the ischemic event in children. Forty-six patients from zero to 18 years of age diagnosed with AIS were studied in the period between March 2002 and September 2003. Laboratory tests were realised including coagulation proteins and echocardiogram. AIS of the newborn occurred in 37% of the cases. Focal seizures and hemiparesis were the most frequent symptoms; 40% of the patients presented prior pathologies. Abnormalities of the S and C proteins occurred in 22% and 17%. Associated alterations, particularly those that generate a hypercoagulability state, indicate more than one risk factor for this disease in childhood. PMID- 15334220 TI - [Anthropometric cranial measures of normal newborn]. AB - OBJECTIVE: The study has been carried out through anthropometric measures of Bahian newborns, to define measures according with the reality of this region. METHOD: Study type transversal cut in term babies with the measures of cephalic perimeter, biauricular and anteroposterior distances, fontanel and cephalic index. RESULTS: 388 babies, 204 (52.6%) of males and 184 (47.4%) females had been evaluated. The cephalic perimeter varied between 31.0 and 38.0 cm, with 34.4 +/- 1.2 average. The cephalic index varied between 0.75 and 1.06, with 0.91 +/- 0.05 average. CONCLUSION: The anthropometric measures are surveyed aiming at precocious detention of anomalies. Possibly, these measures can be influenced by racial factors; however, the measures carried through in Brazil follow values of foreign authors. Thus, the study shows data of a Northeastern region and points out the importance of the accomplishment of multicentric studies. PMID- 15334221 TI - [Risk factors for bacterial meningitis in the newborn]. AB - The aim of this study is to describe the risk factors for bacterial meningitis in newborns, and to analyze the prevalence of these factors, considering or not the low birth weight presence. Fifty newborns with bacterial meningitis were analyzed, excluding the ones with meningomyelocele or congenital infection. In the statistical analysis, the Fisher's exact test was used, considering significant the p < 0.05 values. This study has shown that prematurity, low birth weight and presence of previous infectious diseases in the newborn or in the mother were important risk factors for meningitis. Among low birth weight newborns, invasive procedures, especially tracheal intubation, use of central venous catheter and previous use of antibiotics, were significantly associated to the meningitis occurrence. These results indicate that the improvement in the prenatal care and in the hospital infection control are measures of high importance in the decrease of the incidence of neonatal bacterial meningitis. PMID- 15334222 TI - [Audiologic late prognosis due to meningitis in children]. AB - INTRODUCTION: Auditory deficit has been considered one of the main late manifestations of meningitis, when it occurs during the two first years of life. Few studies have been conducted in Brazil explaining the evolution of children who had meningitis and the percentage, as well the neurological gravity, the auditory problems and sequels after the children left the hospital. OBJECTIVE: To characterize the main auditory and neurological correlation, delineating the profile of the auditory deficits found five years after the central nervous system infection. METHOD: We evaluated children between 5 and 7 years old, admitted in Couto Maia Hospital in the year of 1997, that had been diagnosed with meningitis at an age of two years or below. RESULTS: 19 children passed through the neurological and auditory evaluation. The average age was 6 years and 68.42% were male. In relation to etiology, 52.63% pyogenic, 42.1% viral, 5.26% due to tuberculosis. Auditory alterations had occurred in 26.31% of this population. CONCLUSION: Auditory problems cause academic and social implications in the affected children, especially those of school age. We confirmed the necessity of audiologic following of all children with previous history of meningitis. PMID- 15334223 TI - [Evaluation of some aspects of the acquisition and development of language in pre term born children]. AB - The correction of the age of pre-term infants for the motor evaluation has been the accepted practice but it has not been clear in other areas. This study compared indicators of the acquisition and development of language, considering corrected and chronological ages. Twenty healthy infants born between the 28th and 36th week of gestation (median 32 weeks), weighing 800g to 2380g (median 1590g), 9 AGA and 11 SGA, were followed up to 15 months age. As a reference for normality, evaluation of Costa et al. (1992) was used, which groups predictable behavior in 5 levels. For receptive language, considering the chronological age, normal performance occurred at all levels except for Level I (0-3 months). For expressive language, considering the chronological age, 6 (12%) of the 50 evaluations showed normal performance. With their age corrected, in 16 evaluations (40%) the infants achieved the expected level, mainly at 6 and 12 months age. On the whole, for the chronological age, there was a larger number of AGA with normal performance (p<0.05). We conclude that with the use of the Costa method, it was unnecessary to correct the age for receptive language evaluation, and that, for the expressive, the high frequency of normal results at the corrected ages for 6 and 12 months, suggests that these ages constitute periods of intensification of vigilance. PMID- 15334224 TI - [Diagnosis of children with specific language impairment using a developmental scale]. AB - Specific language impairment (SLI) should have an early diagnosis, since it can interfere with social and school adaptation of the child. The aim of this study was to verify the performance of children with SLI in comparison with normal children using the Behavior Developmental Scale of Gesell and Amatruda. Twenty five SLI children, 3 to 6 years of age, were evaluated. This group was compared to 50 normal children of the same age. Children of control group showed better performance in all aspects of the scale. The medium value of the studied group was borderline in adaptative and social aspects, and was slightly below the medium in the language aspect. We conclude that language disorder may impair the assessment of other areas of development. Nevertheless, this scale may be useful in the evaluation of children with SLI. PMID- 15334225 TI - Balance and motor coordination are not fully developed in 7-year-old blind children. AB - Visually impaired children show difficulties in recognizing their own bodies, objects around then and the spatial parameters that are essential for independent movement. This study analyzes the neuro-psychomotor development of a group of congenitally visually impaired children as compared to children with normal sight. We have evaluated two groups of seven-year-olds by means of neurological evolution examination (NEE). The group studied comprised 20 blind children and the control group comprised 20 children with normal sight, and they were paired up according to age and gender. In some tests, the blind children were guided by touch. The visually impaired children performed worse in tests evaluating balance and appendage coordination compared to normal sighted children (p< 0.001), and this suggests that visual deficiency impairs children's neuro-psychomotor development. PMID- 15334226 TI - [How children aged 7 to 11 explain their headache]. AB - The aim of the present study was to observe the ability of children aged 7 to 11 in describing their headache during anamnesis. For this purpose, two evaluations of 94 children were performed in individual assessments, done by the same Pediatrician, within a six to eight week interval, without the presence of adults. The characteristics of headache in these children could be properly evaluated during the interviews. There were no remarkable conflicting information between the two interviews. Although the headache of the child is usually described by the accompanying adult during the consultation, childhood headache should really be informed by the patient. In this group of young children (7 to 11 years old), information could be obtained without difficulty since we allowed the child enough time and the use of his (her) own words. PMID- 15334227 TI - [Changes in cortical power distribution produced by memory consolidation as a function of a typewriting skill]. AB - The present study aimed to investigate alterations in EEG patterns in normal, right-handed individuals, during the process of learning a specific motor skill (typewriting). Recent studies have shown that the cerebral cortex is susceptible to several changes during a learning process and that alterations in the brain's electrical patterns take place as a result of the acquisition of a motor skill and memory consolidation. In this context, subjects' brain electrical activity was analyzed before and after the motor task. EEG data were collected by a Braintech 3000 and analyzed by Neurometrics. For the statistical analysis, the behavioral variables "time" and "number of errors" were assessed by a one-way ANOVA. For the neurophysiological variable "Absolute Power", a paired t-Test was performed for each pair of electrodes CZ-C3/CZ-C4, in the theta and alpha frequency bands. The main results demonstrated a change in performance, through both behavioral variables ("time" and "number of errors"). At the same time, no changes were observed for the neurophysiological variable ("Absolute Power") in the theta band. On the other hand, a significant increase was observed in the alpha band in central areas (CZ-C3/CZ-C4). These results suggest an adaptation of the sensory-motor cortex, as a consequence of the typewriting training. PMID- 15334228 TI - [Phobic postural vertigo: clinical aspects and course of illness]. AB - Phobic postural vertigo (PPV) is the second most common diagnosis on a dizziness unit. It is a somatoform syndrome characterized as a chronic and incapacitating condition with subjective imbalance and short attacks of dizziness. During a period of 18 months, PPV was observed in 41 patients among 251 of a dizziness unit. Twenty-six had primary PPV, among whom 65% had depressive or anxiety disorders, and 15 patients were diagnosed at secondary PPV. Normal neurological examination and diagnostic tests were observed in most cases. A favorable response to treatment (antidepressants, benzodiazepines, psychotherapy and/or orientation) was observed in 62% of all patients, without difference between both groups -- primary and secondary PPV. Despite the high prevalence, PPV is misdiagnosed. Therefore one must attempt to recognize it, since its appropriate treatment prevents recurrence and incapacitation. PMID- 15334229 TI - [Prevalence of antimicrosomal antibodies in patients with multiple sclerosis]. AB - The aim of this study was to assess the prevalence of Antimicrosomal Antibodies AMA, thyroid function and the occurrence of hypothyroidism symptoms in patients with Multiple Sclerosis (MS). Clinical examination was carried out in 21 MS patients; thyroid-stimulating hormone (TSH), thyroxine (T4), free T4 and AMA were measured. Mean age was 41.05 years. Hypothyroidism symptoms included fatigue, weakness, lethargy and paresthesia. AMA were found in four patients (19%). Three categories of disease duration were considered: <60 months (3 patients AMA+; 7 AMA-), 60-120 months (8 patients AMA-), and >120 months (1 patient AMA+; 2 AMA-). Two patients presented decreased free T4 levels, but there was no associated decrease in T4 and TSH levels. In two patients, a mild increase in TSH levels was observed: one presented normal T4 levels (subclinical hypothyroidism) and the other one had low free T4 levels (classical hypothyroidism). We conclude that AMA measurement and thyroid function tests should become part of the routine assessment of MS patients, in view of the inaccuracy currently observed in the assessment of clinical hypothyroidism as a result of the superposition of hypothyroidism and MS signs and symptoms. PMID- 15334230 TI - [Carotid endarterectomy under regional anesthesia: follow-up of 104 patients]. AB - Cervical carotid stenosis is one of the main causes of ischemic stroke. Carotid endarterectomy is a safe procedure for treatment of moderate and severe symptomatic and asymptomatic carotid stenosis. Regional anesthesia allows neurological evaluation of the patient during the surgery. We reviewed the results of 104 patients operated on at our institution under regional anesthesia during the period of April 1996 and May 2002. 64 patients were symptomatic (61.54%) and 40 asymptomatic (38.46%). All patients had carotid stenosis over 70%. The patients were followed from one to 72 months (Mean: 29.5). Three patients had cervical hematoma, that required surgical drainage. Two patients had minor stroke at the same side of the endarterectomy at the post-operative period and another two during the follow-up (1.92%). Two patients died due to complications related to the surgery (1.92%). Our results, compared with the literature, show that endarterectomy is a safe procedure to treat moderate or severe carotid artery stenosis. PMID- 15334231 TI - [Spontaneous intraparenchymatous hemorrhage: findings at computed tomography]. AB - Computed tomography studies of 250 patients with spontaneous hemorrhage were examined in three hospitals in the city of Rio de Janeiro. The goal of this study was to identify the aspects of this disease that appear most frequently in this type of exam. Deep intracerebral hematomas have had the highest incidence followed by lobar hemorrhage, with thalamus being more frequently affected. Cerebellar hemorrhage was much rarer, with brainstem bleeding observed in few patients. The age group with a peak incidence was at 61 to 70 years. Headache was the most frequently related symptom and elevated levels of blood pressure were found in most of the cases. There was no pronounced difference as to predominance in either sex or side most affected but it was observed that the onset of this disease occurs at an earlier age in men than in women. Blood draining into the ventricular system occurred more frequently in deep hematomas. PMID- 15334232 TI - [Tarlov cysts: report of four cases]. AB - Four perineurial cysts cases (Tarlov's cysts) are reported. The purpose of this study is to describe and to compare them with data from a literature review. The evaluation was performed among 88 adult patients with symptoms of radiculopathy, sacral pain, low back pain. Four patients revealed Tarlov's cysts (4.5%). The diagnosis was made by magnetic resonance imaging. Four cases underwent sacral laminectomy. Following surgery, the claudication pain resolved with no motor or sensory deficits. Tarlov's cysts should be considered as a differential diagnosis of sacral radiculopathy, sacral or lumbar pain syndromes and mainly to the lumbar disc prolapse. The goal of the surgical treatment is to relieve the neural compression and stop bone erosion. PMID- 15334233 TI - [Magnetic resonance imaging in 67 cases of glioblastoma multiforme and occurrence of metastases]. AB - PURPOSE: The purpose of this paper is to demonstrate the main MRI characteristics of glioblastoma multiforme (GBM), the most common CNS primary tumor, emphasizing its location and the occurrence of metastases. METHOD: The MR imaging of 67 pathologically proven cases of glioblastoma multiforme were retrospectively reviewed. The exams were realized in the period between 1995 and 2003, in one of three 1.5 Signa GE units (Milwaukee, WI). RESULTS: The ages of the patients ranged from 4 years to 86 years, mean 60 years, and the occurrence of the tumor was preponderant among men, with 39 cases (58%). The most common location was in the frontal lobes (47%) followed by the temporal lobes (18%) and the parietal lobes (16%). In 19% of the cases there were involvement of more than one site and long distance metastases were seen in 22% of the patients. CONCLUSION: According to the literature, the most common location of GBM was in the frontal lobe of older than 50 years old men. Metastases occurred in 22% of our cases. PMID- 15334234 TI - [Screening of occult spinal dysraphism by ultrasonography]. AB - Ultrasonography for diagnosis of spinal cord diseases has been used since the eighties. There are different fields for its use: traumatic spine lesions, intra operative use in trauma and spine tumors surgery, pre-operative and follow up study for spinal dysraphism, occult spinal dysraphism of the newborn. Patients with suspected occult spinal dysraphism are the most important indication for the method. Ultrasonography was performed in 292 patients of Hospital das Clinicas, Universidade Federal de Minas Gerais. Lesions such as conus medullaris lipomas, low level of conus medullaris, filum terminale tethering, cauda equina cysts and dermal sinuses can be well demonstrated. There is no difficulty in interpreting the image. The exam does not need sedation; it is safe, fast and cheap. Ideally it should be performed by the neurosurgeon as it may provide many information during the examination. The ultrasonography does not replace magnetic resonance imaging but it is an excellent method for screening. PMID- 15334235 TI - [Microsurgical anatomy of the recurrent laryngeal nerve: applications on the anterior approach to the cervical spine]. AB - OBJECTIVE: To present an anatomical description of the recurrent laryngeal nerve (RLN) on both sides of the larynx as it relates to the possible lesion mechanisms in anterior cervical spine surgery. METHOD: Twelve adult cadavers were examined from the microsurgical laboratory at the School of Medicine at UFMG, MG, Brazil. The data collected were analyzed in terms of frequency, average and standard deviation. RESULTS: The left RLN had a total average length of 9.4 +/- 1.6 cm entering the larynx in 36.3% of the cases at the approximate height of C5, 18.2% at C4, 18.2% at C5-C6, 18.2% at C6 and 9.1% at C4-C5. Recurrence appeared in 45.4% of the cases at the approximate height of T3, 18.2% at T3-T4, 18.2% at T4 and 18.2% at T5. The right RLN had a total average length of 5 +/- 0.3 cm entering the larynx in 44.4% of the cases at the approximate height of C5, 44.4% at C6, and 11.1% at C3-C4. Recurrence appeared in 60% of the cases at the approximate height of T1, 30% at C7 and 10% at T2. CONCLUSION: The right RLN was found to be more vulnerable to operational lesions due to two complimentary yet different reasons, trajectory and length. Owing to the fact that a more oblique and unprotected trajectory is not related directly with the tracheoesphageal groove, there is a strong possibility of direct traumas occurring as with the resulting compression from the employ of surgical retractors or an accidental cut, principally in surgeries involving lower vertebral levels. In the same way, the smallest RNL length favors the stretching of fibers during the per-operative traction. PMID- 15334236 TI - [Characteristics of patients with head injury at Salvador City (Bahia--Brazil)]. AB - OBJECTIVE: To describe the characteristics from people with head injury at Salvador city, as well, to point out the most frequent age groups, their disabilities and lethality rates. METHOD: Assessment and notification of 555 medical files from victims with head injury assisted at the General Hospital of Bahia during 2001. RESULTS: 82.9% from all victims were male and 17.1% female; the most frequent age group was 21 to 30 years; the most relevant cause of head injury was traffic accident (40.7%), followed by physical violence with or without weapons (25.4%) and falls (24%). CONCLUSION: The most evident profile group involved in accidents with head trauma implications was young male that usually had traffic injuries, with expressive rates of disabilities and lethality. PMID- 15334237 TI - Transcranial doppler sonography in two patients who underwent decompressive craniectomy for traumatic brain swelling: report of two cases. AB - The role of decompressive craniectomy in the treatment of severe posttraumatic cerebral swelling remains quite a controversial issue. To the best of our knowledge, there is no study demonstrating the effect of decompressive craniectomy on cerebral blood flow (CBF) velocity by means of transcranial Doppler sonography (TCD). We present two patients who developed traumatic brain swelling and uncontrollable intracranial hypertension with coma and signs of transtentorial herniation. One patient underwent bifrontal, while the second, unilateral, frontotemporoparietal decompressive craniectomy with dural expansion. In both patients, TCD examinations were performed immediately before and after surgery to study the cerebral hemodynamic changes related to the operations. Pre and postoperative TCD examinations demonstrated a significant increase in blood flow velocity in the intracranial arteries in both subjects. In conclusion, our cases suggest that decompressive craniectomy with dural expansion may result in elevation of CBF velocity in patients with massive brain swelling. The increase in CBF velocity appears to occur not only in the decompressed hemisphere, but also on the opposite side. PMID- 15334238 TI - Aneurysm growth after brain tumor removal: case report. AB - A rare case of rapid growth of an aneurysm after a posterior fossa meningioma removal in a 69-year-old lady is reported. Serial angiography, cerebral computed tomography and magnetic resonance imaging are presented. The patient harbored risk factors to both aneurysm formation and growth as current cigarette smoking, arterial hypertension, female sex and reduction of intracranial hypertension. One year follow up after the first surgical procedure is presented. PMID- 15334239 TI - Axoval neuropathy as initial manifestation of primary amyloidosis: report of a case submitted to bone marrow transplantation. AB - Amyloidosis is a syndrome characterized by deposition of a highly insoluble protein material in the extracellular space that may affect several organs. It may be generalized and idiopathic (primary amyloidosis). We describe the case of a 48 years-old woman with axonal neuropathy associated with proteinuria, whose final investigation resulted in diagnosis of primary amyloidosis (AL). She was submitted to autologous bone marrow transplantation. We discuss some aspects related to diagnosis of neuropathy and current treatment of AL. PMID- 15334240 TI - [Hallervorden Spatz syndrome: magnetic resonance findings. Case report]. AB - Hallervorden-Spatz syndrome is a neurodegenerative disease, autosomic recessive with two clinical features: early and late onset. Psychiatric, pyramidal and extrapyramidal signs are present in the late subtype. We report the case of a 41 old woman with extrapyramidal signs. Magnetic resonance imaging (MRI) showed the eye-of-the-tiger sign in the medial globus pallidus. This is due to a gliosis (increased signal) and accumulation of surrounding iron (decreased signal intensity) in long TR sequences. There is a strong relationship between MRI findings and the gene mutation responsable for this disease. It makes the MRI sensible for diagnosing this syndrome. PMID- 15334241 TI - [Central pontine and extra-pontine myelinosis in alcoholic patient with Shoshin beriberi]. AB - The central pontine myelinosis is classically related with rapid correction of chronic hyponatremia. Recently, important additional factors have been described in the pathogenesis of this condition. We report the case of a 43-year-old alcoholic malnourished man, with acute renal failure with dialytic treatment, and output failure Shoshin beriberi. He had tetraplegy and coma. Confirmation of pontine and extrapontine myelinosis by magnetic resonance imaging, and abnormalities on auditive evoked potentials are described. PMID- 15334242 TI - [Temporal lobe seizure recorded by magnetoencephalography: case report]. AB - Ictal onset localization is a important factor in presurgical evaluation of epilepsy. This paper describes the localization of a seizure onset recorded by magnetoencephalography (MEG) from a 12-year-old male patient who suffered from complex partial drug-resistant seizures. MRI revealed a 20mm diameter lesion located in left hippocampus. Scalp EEG showed left temporal theta waves. Interictal MEG registrations detected isolated spike-wave activity posterior and inferior to the MRI lesion. Ictal MEG showed continuous spike-wave activity (2 Hz). Dipole localization sited seizure onset in the inferior left temporal gyrus, the same localization of the interictal MEG activity. This ictal activity spreads bilaterally to frontal areas. Intrasurgical electrocorticography recording confirmed interictal MEG results. PMID- 15334243 TI - Laryngeal electromyography in movement disorders: preliminary data. AB - This study describes preliminary laryngeal electromyography (LEMG) data and botulinum toxin treatment in patients with dysphonia due to movement disorders. Twenty-five patients who had been clinically selected for botulinum toxin administration were examined, 19 with suspected laryngeal dystonia or spasmodic dysphonia (SD), 5 with vocal tremor, and 1 with Gilles de la Tourette syndrome (GTS). LEMG evaluations were performed before botulinum toxin administration using monopolar electrodes. Electromyography was consistent with dystonia in 14 patients and normal in 5, and differences in frequency suggesting essential tremor in 3 and Parkinson tremors in 2. The different LEMG patterns and significant improvement in our patients from botulinum toxin therapy has led us to perform laryngeal electromyography as a routine in UNICAMP movement disorders ambulatory. PMID- 15334244 TI - qEEG and dementia. PMID- 15334248 TI - Plumbagin quantification in roots of Plumbago scandens L. obtained by different extraction techniques. AB - The Plumbago genus belongs to the Plumbaginaceae family and it is known due to its variety of biological uses, most of them attributed to the presence of naphthoquinones. Plumbagin is a naturally occurring naphthoquinone that can be obtained from roots of Plumbago scandens L. In order to find out the better technique for plumbagin extraction, were applied: static maceration, dynamic maceration, with assistance of ultrasonic waves and in Soxhlet apparatus. Four compounds were qualitatively detected in all extracts: the naphthoquinones plumbagin and epi-isoshinanolone, palmitic acid and sitosterol. Plumbagin was always the major component in all analyzed extracts and it was quantitatively determined by gas chromatograph coupled with mass spectrometer. Soxhlet was the most efficient extraction technique however, prolonged heating time promoted plumbagin degradation. PMID- 15334249 TI - Chemical constituents from Vellozia graminifolia. AB - From the hexane and ethyl acetate extracts obtained from stems, roots and leaf sheaths of Vellozia graminifolia, a mixture of linear hydrocarbons, a mixture of pentacyclic triterpenes, five monoisoprenylated flavonoids and one labdane diterpene, (-)-ent-3beta-hydroxy-8(17)-labden-15-oic acid, were isolated. The linear hydrocarbons and minor triterpenes were identified in their corresponding mixtures by High Resolution Gas Chromatography (HRGC) and HRGC coupled to mass spectrometry. The major pentacyclic triterpenes and the diterpene were characterized by spectral data, including 2D NMR experiments, and chemical transformations. PMID- 15334250 TI - The T-cell anergy induced by Leishmania amazonensis antigens is related with defective antigen presentation and apoptosis. AB - Leishmania amazonensis is the main agent of diffuse cutaneous leishmaniasis, a disease associated with anergic immune responses. In this study we show that the crude antigen of Leishmania amazonensis (LaAg) but not L. braziliensis promastigotes (LbAg) contains substances that suppress mitogenic and spontaneous proliferative responses of T cells. The suppressive substances in LaAg are thermoresistant (100 degrees C/1h) and partially dependent on protease activity. T cell anergy was not due to a decreased production of growth factors as it was not reverted by addition of exogenous IL-2, IL-4, IFN-gamma or IL-12. LaAg did not inhibit anti-CD3-induced T cell activation, suggesting that anergy was due to a defect in antigen presentation. It was also not due to cell necrosis, but was accompanied by expressive DNA fragmentation in lymph node cells, indicative of apoptosis. Although pre-incubation of macrophages with LaAg prevented their capacity to present antigens, this effect was not due to apoptosis of the former. These results suggest that the T cell anergy found in diffuse leishmaniasis may be the result of parasite antigen-driven apoptosis of those cells following defective antigen presentation. PMID- 15334251 TI - Genes and chromosomes: control of development. AB - The past decade has witnessed immense progress in research into the molecular basis behind the developmental regulation of genes. Sets of genes functioning under hierarchical control have been identified, evolutionary conserved systems of genes effecting the cell-to-cell transmission of transmembrane signals and assigned a central role in morphogenesis have been intensively studied; the concept of genomic regulatory networks coordinating expression of many genes has been introduced, to mention some of the major breakthroughs. It should be noted that the temporal and tissue-specific parameters of gene expression are correctly regulated in development only in the context of the chromosome and that they are to a great extent dependent on the position of the gene on the chromosome or the interphase nucleus. Moreover epigenetic inheritance of the gene states through successive cell generations has been conducted exclusively at the chromosome level by virtue of cell or chromosome memory. The ontogenetic memory is an inherent property of the chromosome and cis-regulation has a crucial role in its maintenance. PMID- 15334252 TI - Effects of insulin and actin on phosphofructokinase activity and cellular distribution in skeletal muscle. AB - In this work, we report evidences that the association of phosphofructokinase and F-actin can be affected by insulin stimulation in rabbit skeletal muscle homogenates and that this association can be a mechanism of phosphofructokinase regulation. Through co-sedimentation techniques, we observed that on insulin stimulated tissues, approximately 70% of phosphofructokinase activity is co located in an actin-enriched fraction, against 28% in control. This phenomenon is accompanied by a 100% increase in specific phosphofructokinase activity in stimulated homogenates. Purified F-actin causes an increase of 230% in phosphofructokinase activity and alters its kinetic parameters. The presence of F actin increases the affinity of phosphofructokinase for fructose 6-phosphate nevertheless, with no changes in maximum velocity (Vmax). Here we propose that the modulation of cellular distribution of phosphofructokinase may be one of the mechanisms of control of glycolytic flux in mammalian muscle by insulin. PMID- 15334253 TI - Pollen record and paleoenvironment of a 4210 years B.P.old sediment in the Bay of Guanabara, Rio de Janeiro, Brazil. AB - Pollen analysis of a sediment sample obtained at 222 cm from the top of a drilling core collected in the Bay of Guanabara, Rio de Janeiro, was used as a tool to obtain more knowledge about pre-historical human living and environment. 14C datation revealed the age of 4210 years B.P. Most frequent pollen grains came from plants like Alchornea (Euphorbiaceae), Celtis (Ulmaceae), Lecythidaceae, Meliaceae, Ochnaceae and spores from forest Pteridophyta. Palynology and environmental studies revealed that agricultural activities could not be detected. The dense tropical rain forest was the dominant vegetation occurring in this region. PMID- 15334254 TI - Glycosynapses: microdomains controlling carbohydrate-dependent cell adhesion and signaling. AB - The concept of microdomains in plasma membranes was developed over two decades, following observation of polarity of membrane based on clustering of specific membrane components. Microdomains involved in carbohydrate-dependent cell adhesion with concurrent signal transduction that affect cellular phenotype are termed "glycosynapse". Three types of glycosynapse have been distinguished: "type 1" having glycosphingolipid associated with signal transducers (small G-proteins, cSrc, Src family kinases) and proteolipids; "type 2" having O-linked mucin-type glycoprotein associated with Src family kinases; and "type 3" having N-linked integrin receptor complexed with tetraspanin and ganglioside. Different cell types are characterized by presence of specific types of glycosynapse or their combinations, whose adhesion induces signal transduction to either facilitate or inhibit signaling. E.g., signaling through type 3 glycosynapse inhibits cell motility and differentiation. Glycosynapses are distinct from classically-known microdomains termed "caveolae", "caveolar membrane", or more recently "lipid raft", which are not involved in carbohydrate-dependent cell adhesion. Type 1 and type 3 glycosynapses are resistant to cholesterol-binding reagents, whereas structure and function of "caveolar membrane" or "lipid raft" are disrupted by these reagents. Various data indicate a functional role of glycosynapses during differentiation, development, and oncogenic transformation. PMID- 15334255 TI - The transition from memory retrieval to extinction. AB - Memory is measured by measuring retrieval. Retrieval is often triggered by the conditioned stimulus (CS); however, as known since Pavlov, presentation of the CS alone generates extinction. One-trial avoidance (IA) is a much used conditioned fear paradigm in which the CS is the safe part of a training apparatus, the unconditioned stimulus (US) is a footshock and the conditioned response is to stay in the safe area. In IA, retrieval is measured without the US, as latency to step-down from the safe area (i.e., a platform). Extinction is installed at the moment of the first unreinforced test session, as clearly shown by the fact that many drugs, including PKA, ERK and protein synthesis inhibitors as well as NMDA receptor antagonists, hinder extinction when infused into the hippocampus or the basolateral amygdala at the moment of the first test session but not later. Some, but not all the molecular systems required for extinction are also activated by retrieval, further endorsing the hypothesis that although retrieval is behaviorally and biochemically necessary for the generation of extinction, this last process constitutes a new learning secondary to the unreinforced expression of the original trace. PMID- 15334256 TI - Improving methods for epidemiological control of canine visceral leishmaniasis based on a mathematical model. Impact on the incidence of the canine and human disease. AB - The mathematical model described by Dye (1996) condemned the epidemiological canine visceral leishmaniasis control campaign, considering it non-efficient. Using this model, we mathematically demonstrate that the control is not efficient, only at low kappa values (rate at which latent and infectious dogs are lost by the destruction program) which match the canine seropositivity observed in the field by the immunofluorescency (IF) blood eluates analysis. With higher k values, corresponding to IF (kappa = 0.07) or ELISA (kappa = 0.25) results in sera samples, the number of infectious dogs declines to a Ro =1 or Ro =0, respectively, interrupting the transmission and the advancement of epidemics. We also experimentally demonstrate that the dog removal, following the results of IF of sera, instead of eluates lead to a 57% (p < 0.005) decrease in canine cases and 87.5% (p < 0.005) in human cases. Our mathematical and experimental results indicate that the control campaign become more efficient by enhancing the sensitivity of the diagnostic assay. PMID- 15334259 TI - Approximate Entropy as a measure of complexity in sap flow temporal dynamics of two tropical tree species under water deficit. AB - Approximate Entropy (ApEn), a model-independent statistics to quantify serial irregularities, was used to evaluate changes in sap flow temporal dynamics of two tropical species of trees subjected to water deficit. Water deficit induced a decrease in sap flow of G. ulmifolia, whereas C. legalis held stable their sap flow levels. Slight increases in time series complexity were observed in both species under drought condition. This study showed that ApEn could be used as a helpful tool to assess slight changes in temporal dynamics of physiological data, and to uncover some patterns of plant physiological responses to environmental stimuli. PMID- 15334260 TI - [Clinical differences observed in patients with dengue caused by different serotypes in the epidemic of 2001/2002, occurred in Rio de Janeiro]. AB - The authors evaluated clinical and epidemiological differences among the serotypes of dengue in Rio de Janeiro's 2001-2002 outbreak of the disease. Out of 362 cases that had viral isolation samples, notified by the Information System for Notification Diseases (SINAN), from January/2001 to June/2002, 62 were caused by serotype 1,62 by serotype 2 and 238 by serotype 3. In comparison with serotype 2, an individual infected by serotype 3 had a 6.07 times higher chance (OR = 6.07; CI: 1.10-43.97) of presenting shock and a 3.55 times higher chance (OR = 3.55; CI: 1.28-9.97) of developing exanthema. When compared to serotype 1, serotype 3 had a 3.06 times higher chance (OR = 3.06; CI: 0.99-9.66) of causing abdominal pain and a 3.61 times higher chance of exanthema (OR = 3.61; CI: 1.16 11.51). It was found that individuals infected by serotype 3 of the virus presented signs indicating a more severe disease. PMID- 15334261 TI - [The epidemic of dengue and hemorrhagic dengue fever in the city of Rio de Janeiro, 2001/2002]. AB - The following study was intended to evaluate the occurrence of typical signs and symptoms in the cases of classic dengue and hemorrhagic dengue fever, during the 2001-2002 epidemic in the city of Rio de Janeiro. The authors reviewed 155,242 cases notified to the Information System of Notification Diseases, from January/2001 to June/2002: 81,327 cases were classified as classic dengue and 958 as hemorrhagic dengue fever, with a total of 60 deaths. Common symptoms, such as fever, headache, prostration, myalgia, nausea and retro-orbital pain, had a high incidence in both classic and hemorrhagic dengue fever. On the other hand, hemorrhagic signs and other signs of severe disease, such as shock, gastrointestinal bleeding, petechiae, epistaxis, abdominal pain and pleural effusion, were strongly associated to hemorrhagic dengue fever. Besides, the occurrence of death was 34.8 times higher in hemorrhagic dengue fever than in classic dengue (OR = 34.8; CI 19.7-61.3). PMID- 15334262 TI - [Clinical and epidemiological aspects of visceral leishmaniasis in children up to 15 years of age in Alagoas, Brasil]. AB - In order to investigate epidemiological and clinical characteristics of visceral leishmaniasis in children up to 15 years old, a prospective study was carried out in Alagoas, Brasil from 1981 to 1995. Of the 530 diagnosed cases, predominantly from the rural area of Alagoas State, 58% were male and 42% female, being 55.3% children under 5 years old. The most frequently observed clinical manifestations were: hepatosplenomegaly, fever and parlor. The average size of the liver and the spleen of patients with shorter time of disease (<30 days) were smaller than those presenting sickness for a extended time (>or= 360 days). No matter the length of disease there was reduction of the liver and the spleen after treatment. However, the reduction of the spleen was higher in those patients with less time of sickness. With relation to liver that difference was not observed. PMID- 15334263 TI - [Visceral leishmaniasis among Indians of the State of Roraima, Brazil: clinical and epidemiologic aspects of the cases observed from 1989 to 1993]. AB - A description of the epidemiological profile of visceral leishmaniasis among Indians in the State of Roraima, Brazil, was based on the clinical characteristics of human and dog disease, ecological aspects of the area where the cases occurred and entomologic investigations performed from 1989 to 1993. The 82 human cases were reported in six out of eight Counties that existed then in the state; there was a 69.5% predominance of male cases among those registered and a greater (52.4%) occurrence of the disease in children from zero to ten years old. The rate of natural infection was 10.3% out of 3,773 dogs examined in 74 different locations. Lutzomyia longipalpis was found in 31 areas with greater prevalence of the disease. The human and animal cases as well as the vectors were concentrated in areas where mountains and arable soil predominate, typical locations for the occurrence of American visceral leishmaniasis. PMID- 15334264 TI - [The AIDS epidemic in Brazil, 1991-2000: space-time description]. AB - The incidence of AIDS was described by an ecological study of Brazilian municipalities, considering cases reported from 1991 to 2000. An increasing incidence was observed in women and among individuals with a low educational level. Sao Paulo State had different epidemiological profiles when the State Capital was compared to other cities, with an overall 46% of reported cases in the Country; Pernambuco State had almost the same profile since the beginning of the epidemic in Brazil; In Santa Catarina State the category of intravenous drug users had the greatest exposure. The increasing number of cases reported under the category of unknown transmission showed the increased bias in measurement by the surveillance system. PMID- 15334265 TI - [Association between Vibrio cholerae and zooplankton of estuaries of Sao Marcos Bay/Sao Luis - MA, Brazil]. AB - This study was carried from October 1997 to October 1998 with the purpose of investigating the link between the species Vibrio cholerae and zooplankton in the estuaries of rivers Anil and Bacanga in Sao Luis, Maranhao. Research of viable but non culturable forms of Vibrio cholerae O1, and analyze the correlation between pH values, salinity and water temperature with the presence of bacteria in zooplankton samples. The traditional method of cultivation and fluorescent antibody technique were applied to detect the bacteria. A total of 52 samples of zooplankton were collected. There was a predominance of Copepoda. The culture enabled 55 isolates of Vibrio cholerae non-O1. The serogroups O1 and O139 were identified in 37 (71.1%) and 17 (32.7%) samples respectively by the fluorescence. Viable but non culturable forms of Vibrio cholerae were detected in 70.8% of the samples studied. A significant correlation was established between salinity and the water's pH value and Vibrio cholerae. PMID- 15334266 TI - [Molecular study of Vibrio cholerae non-O1 isolated from zooplankton of Sao Marcos Bay/Sao Luis - MA, Brasil]. AB - The study was developed to analyze the plasmidial DNA, research virulence genes and identify genetic diversity of 31 strains of Vibrio cholerae non-O1 isolated from zooplankton of the Bacanga and Anil rivers in Sao Luis-MA. The study of plasmidial DNA showed 2 or 3 plasmids from 10 strains between 5.5 and 40 kilobasis. There was only single ribotype pattern. PCR methods did not show the genes ctxA, ace and zot, while RADP-PCR identified genetic diversity in the strains, showing the potential for variability in this species. PMID- 15334267 TI - Establishment of HTLV-I-infected cell lines from peripheral blood mononuclear cells of Brazilian patients. AB - To investigate epidemiological and pathogenetic features of HTLV-I infection, a cohort of carriers has been followed at the USP Teaching Hospital since 1991. This study describes the establishment of cell lines from peripheral blood mononuclear cells (PBMC) of infected subjects. Ex vivo PBMC were cultured with those from a seronegative donor and morphologic evidence of cell transformation was obtained after 90 days with detection of multinucleated cells exhibiting cerebriform nuclei. Integration of HTLV-I proviral DNA and expression of viral antigens was demonstrated in culture by PCR and immunofluorescence. Cell lines were maintained for 240 days, gradually weaned from exogenous IL-2. Immunophenotyping of cell lines on flow cytometry yielded evidence of cell activation. Establishment of HTLV-I-infected cell lines from ex vivo PBMC is feasible and may be useful for studies on lymphocyte phenotypic changes and on mechanisms of HTLV-induced cell proliferation. Moreover they may be used with diagnostic purposes in immunofluorescence tests. PMID- 15334268 TI - Magnetic resonance imaging and ultrasound in hepatosplenic schistosomiasis mansoni. AB - We report the findings of abdominal ultrasound and magnetic resonance imaging observed in a patient with advanced schistosomiasis mansoni. A 25-year-old man with hepatosplenic schistosomiasis and variceal bleeding confirmed by upper endoscopy was submitted to abdominal ultrasound and magnetic resonance imaging. During surgery for portal hypertension, a liver biopsy was taken and the diagnosis of Symmers' fibrosis was confirmed. magnetic resonance imaging scans gave more precise information about the gallbladder, periportal thickening and abdominal venous system than did the ultrasound. PMID- 15334269 TI - [Epidemiologic aspects of tuberculosis in the Surui Indians, Brazilian Amazon]. AB - Tuberculosis persists as a serious public health problem in Brazil. Prevalence rates are alarming in certain social groups, including indigenous peoples. This article presents an epidemiological analysis of records for the Surui Indians available at the Tuberculosis Control Program in the Municipality of Cacoal, Rondonia. The study includes a descriptive statistical analysis of cases reported from 1975 to 2002. There is evidence that the Surui have an increased risk of acquiring and dying from tuberculosis as compared to other indigenous groups in Rondonia as well as non-Indians. The average incidence coefficient for tuberculosis in the Surui was 2518.9 per 100,000 inhabitants in the period 1991 2002. It was observed that 45% of the cases were diagnosed in children < 15 years old. Over half of the cases (63.3%) were reported in men. Only 43.2% of the cases were confirmed by sputum microscopy. The use of PPD skin tests, histopathological exams or bacteriological culture were not reported throughout the period. Attention is called to the need for prevention and control measures specifically tailored to the reality of indigenous peoples. PMID- 15334270 TI - [Dengue prevention: focus on information]. AB - The first three months of 2002 witnessed a dengue epidemic in the State of Rio de Janeiro. At that time, health authorities encouraged community participation in the elimination of vector breeding sites. For this campaign, a great quantity of information about the disease was extensively publicized in order to guide the population's preventive action. This paper analyzed the three pamphlets most widely distributed at that time in Rio de Janeiro city considering that this information contributed to the construction of disease representation and its prevention. It was observed that even though this information was provided repeatedly it must be revised. PMID- 15334271 TI - Polymerase chain reaction and restriction fragment length polymorphism analysis of the ITS2 region for differentiation of Brazilian Biomphalaria intermediate hosts of the Schistosoma mansoni. AB - We sequenced the internal transcribed spacer 2 of the ribosomal DNA (ITS2-DNAr) from the three Schistosoma mansoni intermediate hosts in Brazil: Biomphalaria glabrata, Biomphalaria tenagophila and Biomphalaria straminea. Analysis of a restriction map from those sequences allowed us to select putative restriction enzymes able to identify the snail species under study. Four restriction enzymes were used and HpaII provided simple species-specific profiles easily visualized in polyacrylamide gels. The use of ITS2 is advantageous as it provides a small fragment of 460 bp which may be easily amplified by PCR. In the current work, we showed that the amplification of ITS2-DNAr together with HpaII enzyme restriction is an auxiliary molecular tool for the morphological identification of such snails as well as for taxonomic and phylogenetic studies of neotropical planorbids. PMID- 15334272 TI - [Blastocystosis: a high proportion of cases found in schoolchildren of Sao Paulo, State of Sao Paulo, Brazil]. AB - As a part of medical assistance activities, parasitological examination of fecal samples from 227 school children from a public institution of Sao Paulo (SP) revealed a rather high proportion of results positive for Blastocystis hominis. Other protozoan and worm species were markedly scarcer, a peculiar situation according to our judgement. It is acknowledged that blastocystosis is still largely an indefinite and controversial subject, which deserves adequate analysis to avoid drawbacks in the sphere of action of public health and general medical assistance. PMID- 15334273 TI - Recurrent oedema associated with the co-formulation of lopinavir and ritonavir. PMID- 15334274 TI - Pulmonary strongyloidiasis. PMID- 15334275 TI - Psychosocial impact of acne vulgaris: evaluating the evidence. AB - This paper reviews current evidence presented by recent studies on the impact of acne on psychosocial health. Study methodologies, including case-control and cross-sectional surveys, have demonstrated psychological abnormalities including depression, suicidal ideation, anxiety, psychosomatic symptoms, including pain and discomfort, embarrassment and social inhibition. Effective treatment of acne was accompanied by improvement in self-esteem, affect, obsessive-compulsiveness, shame, embarrassment, body image, social assertiveness and self-confidence. Acne is associated with a greater psychological burden than a variety of other disparate chronic disorders. Future studies with a longitudinal cohort design may provide further validation of the causal inference between acne and psychosocial disability provided by the current literature. PMID- 15334276 TI - Ciclopirox (Loprox) gel for superficial fungal infections. AB - Ciclopirox (Loprox) is a broad-spectrum antifungal medication that also has antibacterial and anti-inflammatory properties. Its main mode of action is thought to be its high affinity for trivalent cations, which inhibit essential co factors in enzymes. Clinical trials have shown that ciclopirox gel is a successful treatment for seborrheic dermatitis of the scalp as well as for tinea pedis. Adverse effects are generally mild and include a skin-burning sensation, contact dermatitis, and pruritus. Ciclopirox is indicated in the US for the treatment of tinea pedis, tinea corporis, pityriasis versicolor, seborrheic dermatitis, and cutaneous candidiasis. PMID- 15334277 TI - Ablative laser resurfacing -- postoperative care. AB - Wound care after laser skin resurfacing (LSR) is critical for achieving a successful result. The superficial thermal injury created by LSR heals more quickly and with a reduced risk of scarring under occlusion. While open and closed wound care regimens can be employed to expedite reepithelialization, closed methods with semi-occlusive dressings may decrease morbidity. Effective medications and management techniques can help to minimize expected effects of the procedure such as crusting, discomfort, pruritus, erythema, and swelling. PMID- 15334278 TI - Topical agents used in the management of hyperpigmentation. AB - Disorders of hyperpigmentation are difficult to treat, particularly in dark skinned individuals. The goal is to reduce the hyperpigmentation without causing undesirable hypopigmentation or irritation in the surrounding normally pigmented skin. The psychosocial impact caused by these disorders must be considered. Although there are many effective therapeutic modalities available, there are potentially significant side-effects associated with treatment. The most commonly used treatment is topical hydroquinone. There are other phenolic agents, such as N-acetyl-4-cystaminylphenol (NCAP), that are currently being studied and developed. The non-phenolic agents, which include tretinoin, adapalene, topical corticosteroids, azelaic acid, arbutin, kojic acid, and licorice extract, are also used for hyperpigmentation disorders. PMID- 15334279 TI - Ciclopirox shampoo for treating seborrheic dermatitis. AB - Seborrheic dermatitis is a common inflammatory skin disease, affecting between 1% and 3% of immunocompetent adults. While its cause is unknown, a number of predisposing factors have been reported, including the implications of Malassezia yeasts. Various treatment options are available, such as ciclopirox shampoo, which combines anti-Malassezia activity with an anti-inflammatory action. This agent has been shown to be an effective and safe treatment for seborrheic dermatitis of the scalp. PMID- 15334280 TI - Face-lifting: an overview. AB - Numerous adaptations of face-lift techniques have been devised, and each surgical approach has its own risks and benefits, as well as proponents and detractors. All of the conventionally accepted techniques achieve removal of redundant skin. However, its the variations in approach to the deeper soft tissue structures that separate the many face-lifting procedures. A skin only face-lift was the earliest form of surgical rhytidectomy, but failed to achieve significant long-term benefit. Cosmetic surgeons of various backgrounds thus sought to achieve a more durable benefit from the surgery. While certain techniques have been classically ascribed to a particular surgeon, innovation does not occur in a vacuum and many surgeons are responsible for our current state of knowledge with regards to facial rhytidectomy surgery. PMID- 15334281 TI - Evidence-based medicine and healthcare: advancing the practice. PMID- 15334282 TI - Philippe Pinel (1745-1826): liberator of the insane. PMID- 15334283 TI - What is this thing called EBM? PMID- 15334284 TI - Public attitudes to emergency medical services in Singapore: EMS day 2002. AB - INTRODUCTION: In the year 2002, the Society for Emergency Medicine in Singapore Chapter of Paramedics organised the first emergency medical services (EMS) day, to educate the public about basic emergency response skills and to increase public awareness of the local EMS System. METHODS: This was an observational, cross-sectional study. A survey was conducted to find out about the knowledge and attitudes of the public and paramedic volunteer instructors towards the local EMS System. RESULTS: Two hundred and six (81.4 percent) out of 253 members of the public and 70 paramedics (100 percent) responded. For the public, the majority were females (86 percent), mean age (standard deviation [sd]) was 15.9 (7.9) years, range 11.0 to 67.0 years. For the paramedics, mean (sd) age was 26.6 (3.8) years. 61.9 percent were females. The public showed good knowledge of the emergency ambulance number and the indications for calling an ambulance. Public expectations of ambulance response times were significantly shorter than paramedics. They were also less comfortable with ambulance crews performing advanced life support interventions compared with the paramedics. CONCLUSION: Continuing efforts should be made to increase public awareness of the EMS system as well as to manage public expectations regarding response times and the roles of paramedics. EMS day represents one such opportunity. PMID- 15334285 TI - The majority of hepatitis B carriers are not on regular surveillance in Singapore. AB - INTRODUCTION: Carriers of hepatitis B virus (HBV) are at risk of developing long term complications. Regular surveillance helps detect treatable chronic hepatitis, cirrhosis and liver cancer, and is recommended by practice guidelines in the United States, Europe and Singapore. However, there have been few studies evaluating the follow-up of HBV carriers. This study seeks to determine the proportion of HBV carriers on regular follow-up in Singapore and the impact on hepatitis B disease. METHODS: An advertisement was taken in local newspapers advertising for free screening to HBV carriers. 387 persons answered the advertisement. The screening comprised history-taking, physical examination, blood tests (liver panel, alphafoetoprotein, hepatitis B surface antigen (Ag) and hepatitis B eAg) and ultrasonography of liver. Further evaluation was conducted if the screening results were abnormal. RESULTS: Of the 387 HBV carriers, 346 (89 percent) were male and 375 (97 percent) were Chinese. Their mean age was 39 years (range 20-60 years) and 36 percent were positive for HBeAg. 247 (64 percent) were not on regular screening over the past 12 months. 19 (5.4 percent) patients were diagnosed to have complications, namely: 13 had HBeAg-positive chronic hepatitis, two had HBeAg-negative chronic hepatitis, one had early liver cancer who recovered well after a curative resection and three had compensated cirrhosis. CONCLUSION: Our screening programme diagnosed 5.4 percent of complications among 387 asymptomatic HBV carriers. However, 64 percent of the study subjects were not screened regularly and may pose an important public health threat if they develop long-term complications. Further studies are needed to evaluate and improve patient compliance for screening. PMID- 15334286 TI - Immunogenicity of low-dose and conventional-dose recombinant hepatitis B vaccines in healthy adolescents in India. AB - INTRODUCTION: The recombinant hepatitis B vaccine, at a conventional dose of 20 microgrammes, is highly effective in a majority of the population. Recent studies have reported that a 10-microgramme dose is as effective as the conventional dose, especially in young adolescents. This study compares the effect of two different doses of recombinant hepatitis B vaccine (Enivac HB) in healthy adolescents. METHODS: Ninety-two adolescents were randomised into two equal groups to receive either 10 microgrammes (group 1) or 20 microgrammes (group 2) of vaccine using the vaccination schedule of 0, one and six months. Blood samples were collected at 30, 60, 90, 180 and 210 days to detect anti-HBs antibodies. At each interval, geometric mean titres were calculated for seroconverted subjects. RESULTS: After the initial doses of the vaccine, greater proportion of subjects receiving the higher dose seroconverted. However, at six months, all subjects who received the low dose seroconverted, as did those who received the conventional dose. Furthermore, there were significant differences in the anti-HBs geometric mean titres for seroconverters at 180 days (218.27 versus 111.43) and 210 days (345.14 versus 133.35). The difference in the overall reactogenicity for the two dose levels was not remarkable, although the higher dose produced more local symptoms. CONCLUSION: 10-microgramme recombinant vaccine can be used in routine immunisation in healthy adolescents. If quick immunisation is needed, as in high risk groups, the conventional dose should be administered, as it results in earlier seroconversion with higher anti-HBs geometric mean titres. PMID- 15334287 TI - Smoking cessation programme: the Singapore General Hospital experience. AB - INTRODUCTION: The National Health Survey in Singapore reported that the prevalence of smoking had decreased from 20 percent in 1984 to 15 percent in 1998. This may be due to the efforts of smoking cessation education established island-wide. In this study, we review the efficacy of the Singapore General Hospital smoking cessation programme and examine the efficacy of different treatment modalities. METHODS: We studied the immediate quit rate and point prevalence abstinence rates at six and 12 months in our telephone survey. Subjects were patients who attended our programme from June 1999 to December 2002. Pharmacotherapeutic aids utilised with counselling sessions were individualised. RESULTS: The study populations for outpatient and inpatient arms were 394 patients and 425 patients, respectively. In the outpatient programme, mean age was 46 years (range of 12 to 80 years), and the ratio between males and females was 8.6. The outpatient immediate quit rate was 33 percent, and the six and 12 month quit rates were both 36 percent. However, in the inpatient programme, mean age was 65 years (range of 15 to 93 years), and the ratio between males and females was 4.9. The six and 12 month quit rates of the inpatient arms were 30 percent and 32 percent, respectively. Although there is no statistically significant difference in the different treatment modalities, the immediate quit rates for bupropion only and counselling only were relatively higher (36 percent and 41 percent, respectively). These were sustained at more than 35 percent at six and 12 months follow-up. We achieved comparable efficacy compared to published data. Counselling, as a sole therapy, can be effective in a select patient group. One-time inpatient counselling achieved a quit rate (32 percent at 12 months) far superior to previously-reported self-quit rate (3 percent and 8 percent at 12 months). CONCLUSION: We strongly recommend that all inpatients who are smokers to be routinely referred for counselling. PMID- 15334288 TI - Endometriosis of the appendix presenting as acute appendicitis. AB - While endometriosis is fairly common, endometriosis of the appendix is a very rare occurrence. Correct pre-operative diagnosis is uncommon and definitive diagnosis is established by histology of the appendix. We present a case of endometriosis of the appendix that manifested as acute appendicitis in a 40-year old woman. PMID- 15334289 TI - Ectopic thyroid tissue in the parotid salivary gland. AB - Benign ectopic thyroid tissue within the parotid salivary gland is very rare. A 32-year-old woman presented with a slowly-growing, painless mass in the parotid region. The mass, which was clinically diagnosed as a parotid tumour, was found at surgery to be cystic in nature. Histological examination showed thyroid tissue with secondary changes in the cyst wall and colloid in the lumen. On iodine isotope scan, the thyroid gland was found in its normal location. The possible origin of the ectopic thyroid tissue in the parotid salivary gland could be due to a common evolution of the thyroid and parotid glands, a heteroplasia or a metaplasia. PMID- 15334290 TI - Imaging of tuberculosis of the spine. AB - Tuberculosis (TB) is the leading cause of death worldwide that can be attributed to a single infectious agent. With the onset of the AIDS epidemic, there has been a resurgence of TB in recent years. Skeletal TB constitutes 1% to 3% of extrapulmonary cases, and typically involves the spine. TB of the spine should be considered in the differential diagnosis of many spinal conditions affecting patients of all ages. The pathophysiology, clinical and imaging features of TB of the spine are reviewed, with illustrations of findings on radiography, computed tomography and magnetic resonance imaging. Familiarity with the imaging features of TB of the spine may enable a more rapid diagnosis to be made, thereby preventing a delay in diagnosis with its consequent complications. PMID- 15334292 TI - 2001 survey on primary medical care in Singapore. PMID- 15334296 TI - Society of Nuclear Medicine--51st annual meeting. Therapeutic agents. PMID- 15334297 TI - Society of Nuclear Medicine--51st annual meeting. Imaging/diagnostic agents. PMID- 15334298 TI - Society of Nuclear Medicine--51st annual meeting. Neuroimaging. PMID- 15334299 TI - Global cancer event--SMi conference. Clinical Trials in Cancer. PMID- 15334300 TI - Pain therapeutics--fifth annual SMi conference. PMID- 15334301 TI - Prostate cancer--Fourth International Congress. PMID- 15334302 TI - Neurodegenerative disorders--4th annual SMi conference. AB - The prevalence of neurodegenerative diseases in the Western World and Japan is predicted to increase sharply as a consequence of the rapidly aging population. To discuss the development of therapeutics in this increasingly important field, the 2-day Neurodegenerative Disorders SMi meeting once again brought together delegates from both large pharmaceutical companies and smaller technology and analysis companies. The majority of presentations focused on Alzheimer's disease, one of the gravest of such conditions that affects approximately half of all people aged 90 years, and on Parkinson's disease. As well as disclosing new therapeutic targets and preclinical and clinical data for drugs in development, several presentations discussed the development of surrogate markers and imaging techniques to monitor disease progression and therapeutic efficacy. PMID- 15334303 TI - International symposium on nitric oxide, cytokines and inflammation. AB - This meeting was presented by the Fundacao Oswaldo Cruz in conjunction with the University of Calgary and the University of California Los Angeles. The conference focused on novel aspects of the contribution of nitric oxide and cytokines to inflammatory processes, particularly the resolution of inflammation. Sessions featured lectures and symposia by many of the world's leading authorities on the basic science and pharmacology of inflammation, as well as lively, high-standard poster presentations by international scientists, including many young Brazilian researchers. One of the main topics was elucidation of the basic mechanisms and roles of cytokines in inflammation. Moreover, the therapeutic potential of new chemical entities (some derived from snake venoms) and drug targets was highlighted in symposia devoted to the resolution of inflammation, airway inflammation, protease-activated receptors and annexin-1. This report concentrates on potential novel therapeutic agents and targets in this complex and ever-evolving field, although many interesting presentations raised the possibility of new applications for drugs already in the therapeutic arena. PMID- 15334304 TI - Medicinal chemistry--29th National Symposium. SERMs and SARMs. PMID- 15334305 TI - Medicinal chemistry--29th National Symposium. Kinase inhibitors. PMID- 15334306 TI - Prodrugs: effective solutions for solubility, permeability and targeting challenges. AB - This 2-day inaugural conference on prodrugs was presented by Pharmaceutical Education Associates and covered recent developments in prodrug techniques to solve delivery and targeting issues in drug discovery and development. The speakers were drawn from industry and academia, and the conference was attended mostly by researchers working in the pharmaceutical and biotechnology industries. A number of topics were presented at the conference, from basic prodrug design and functional group considerations to drug metabolism involving cytochrome P450 enzymes, from increasing water solubility, bioavailability, permeability and stability to tumor targeting, from the development of new anti-inflammatory agents to anti-HIV agents, and from the use of transporters and receptor-mediated endocytosis in prodrug delivery to the use of gene therapy for enzyme delivery to cancer cells and tissues. Several case studies were presented including improved pharmaceutical products in the clinic and at various stages of development. PMID- 15334307 TI - Drug transport and drug delivery--the Midnight Sun meeting. AB - The Midnight Sun Meeting on Drug Transport and Drug Delivery was held on the island of Tromso in northern Norway, where the sun does not set for 2 months during the summer. The meeting was hosted by the University of Tromso's newly established Institute of Pharmacy and the Controlled Release Society (Nordic Chapter). The meeting, attended by approximately 80 delegates from across Europe, showcased recent advances in drug transport through biological barriers, solid state pharmaceuticals and particulate drug delivery systems. This report will focus on the particulate and solid-state pharmaceuticals sessions, in which lectures were given to demonstrate the benefits in cognitive function associated with omega-3 fish oils, the increase in drug release rates observed on the processing-induced deformation of tablet granules, and the size of polymeric particulates being directly and linearly related to the molecular weight of a polymer. The meeting was held as a single-session event, giving delegates the opportunity to attend all presentations. There was a small poster and exhibitor display, and the meeting attracted sponsorship from a number of companies, namely Polypure AS, Weifa AS, ProBioNeutraceuticals AS, Lipoid GmbH, Clavis Pharma AS and Thermometric AB. PMID- 15334308 TI - Recent progress in research on peroxisome proliferator-activated receptor alpha selective ligands. AB - The understanding of the functions of the nuclear receptor peroxisome proliferator-activated receptor a (PPARalpha) as a regulator of lipid and lipoprotein homeostasis, and the rapid development of parallel high-throughput screening assays to evaluate the activity toward other PPAR subtypes (PPARdelta and PPARgamma), have provided an opportunity to develop novel PPARalpha selective, PPARalpha/gamma dual and PPAR pan agonists for the treatment of various metabolic diseases. This review focuses on the molecular pharmacology of PPARalpha, and summarizes recent literature and patent applications disclosing medicinal chemistry strategies to identify new PPARalpha-selective agonists. The species selectivity of some classes of PPARalpha-selective agonists in response to in vitro PPARalpha transactivation activity is also reported. PMID- 15334309 TI - The pivotal role of drug metabolism and pharmacokinetics in the discovery and development of new medicines. AB - An appropriate drug metabolism and pharmacokinetic (DMPK) profile remains a major hurdle to reducing risk and improving productivity in pharmaceutical R&D, accounting for approximately 40% of all drug failures. For orally administered drugs, failure is often attributable to low intestinal absorption and/or high clearance, causing poor and variable bioavailability. Additional reasons for failure include drug-drug interactions and the presence of active metabolites. With a poor pharmacokinetic profile, it can be difficult to achieve the dose profile required for therapeutic efficacy. The main role that DMPK plays in drug discovery is therefore the prediction of drug metabolism and pharmacokinetics in humans. Successful prediction can be expected to reduce the rate of attrition during drug discovery and development. This has led to the recognition that DMPK is an essential component of the drug discovery process. Both this and the need to screen ever greater numbers of compounds have led to major changes in both technology and the process of drug discovery. PMID- 15334310 TI - Transcription factors in asthma and COPD. AB - Inflammation is a central feature of asthma and chronic obstructive pulmonary disease (COPD) and both are characterized by an increased transcription of pro inflammatory proteins (eg, cytokines, chemokines, growth factors and enzymes). Changes in inflammatory gene transcription are regulated by transcription factors that may therefore play a key role in the pathogenesis of asthma and COPD by amplifying and perpetuating the inflammatory process, and thereby contributing to disease severity and responsiveness to treatment. Several new compounds based on interactions with specific transcription factors or their activation pathways are now in development for the treatment of asthma and COPD. PMID- 15334311 TI - TLK-286. AB - Telik Inc is developing TLK-286, a modified glutathione analog prodrug, for the potential treatment of cancer. PMID- 15334312 TI - Coagulation and fibrinolysis in individuals with advanced liver disease. AB - An overview of normal coagulation and fibrinolysis is presented. The abnormalities of coagulation and fibrinolysis seen in individuals with advanced liver disease is reviewed. The necessary steps in the management of bleeding in patients with advanced liver disease is outlined. Finally those liver diseases associated with vascular thrombosis are identified. PMID- 15334313 TI - Nitric oxide and renal functions in liver cirrhosis. AB - BACKGROUND/AIMS: Nitric oxide, a potent vasodilating agent, has been proposed to play a role in pathogenesis of ascites and hepatorenal syndrome. The aim of this study was to evaluate the interaction between the plasma nitric oxide, nitric oxide synthetase levels and renal functions in patients with different degrees of chronic liver disease. METHODS: The study population included 38 subjects: 14 patients with chronic hepatitis, 11 with preascitic cirrhosis and 13 with ascitic cirrhosis. Nitric oxide and nitric oxide synthetase were determined by colorometric assay. We calculated glomerular filtration rate and fractional sodium excretion. RESULTS: Nitric oxide levels in groups were as follows: 79.28+/ 24.86, 99.03+/-21.31, 197.05+/-49.61 microm, respectively. Nitric oxide synthetase levels were 2.64+/-0.56, 3.64+/-0.89, 7.75+/-2.46 micromol/L/sec, respectively. Nitric oxide and nitric oxide synthetase levels in the ascitic cirrhotic group were significantly higher than in the others (p<0.05). When glomerular filtration rates were compared, the only significant difference was determined between the groups with chronic hepatitis and ascitic cirrhosis (92.31+/-25.21, 48.46+/-16.45, p<0.05). Fractional sodium excretion was significantly increased in the ascitic cirrhotic group (4.42+/-2.76, p<0.05). CONCLUSIONS: Nitric oxide and nitric oxide synthetase increased with progression of liver disease, especially in ascitic cirrhosis. We also showed that this increase negatively affects the renal tubular and glomerular functions. PMID- 15334314 TI - Laryngopharyngeal reflux in laryngeal cancer. AB - BACKGROUND/AIMS: Gastropharyngeal or laryngopharyngeal reflux is considered as a factor in various diseases of the larynx and pharynx. The most important consequence of the reflux into the larynx is laryngeal cancer. METHODS: In this prospective study the incidence of gastropharyngeal and laryngopharyngeal reflux in 22 patients with untreated laryngeal cancer was investigated with 24-hour, double probe pH measurements. A group of 25 patients with heartburn and dyspepsia complaints in whom esophagogastroscopy revealed no pathology and for whom 24-hour pH measurement was indicated served as a control group. RESULTS: Two of the 22 patients never smoked and two others had quit smoking 16 and 25 years previously. All four of these patients revealed gastropharyngeal reflux. In total, 14 of the laryngeal cancer patients (63.6%) revealed gastropharyngeal reflux. The rate of gastroesophageal reflux was close among the two groups (50% in the cancer group vs 32% in the control group, p>0.05), but the laryngopharyngeal reflux rate was much higher in the cancer group (63.6% of the cancer patients vs 20% in the control group, p: 0.003). Among the reflux-positive patients and the controls, gastroesophageal reflux rate was higher in the supine position in cancer patients (12.10% vs 6.25, p: 0.02). In the upright position, control cases revealed higher rates of gastroesophageal reflux than the cancer patients. Laryngopharyngeal reflux rates were slightly higher in laryngeal cancer patients than the controls in both upright (9.29% vs 7.67%, p: 0.6) and supine positions (4.83% vs 3.50%, p: 0.6). CONCLUSIONS: Laryngeal cancer patients and patients with heartburn complaints all have a high rate of gastroesophageal reflux. But cancer patients reveal a higher rate of laryngopharyngeal reflux than the symptomatic patients with normal laryngeal findings. PMID- 15334315 TI - Accuracy of endoscopic ultrasonography in upper gastrointestinal submucosal lesions. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the accuracy of endoscopic ultrasonography (EUS) and consistency of the EUS findings with histopathologic examination. METHODS: EUS was performed in 90 patients with upper gastrointestinal tract submucosal tumors, followed in Turkiye Yuksek Ihtisas Hospital, Gastroenterology Clinic. Histopathological diagnosis and EUS findings of 25 of 90 patients were compared. RESULTS: 48.9% of the lesions were found to have arisen from muscularis propria, 33.3% from submucosa, 6.6% from mucosa and 10% from muscularis mucosae, and 1.2% from serosa of the 90 patients. In 25 patients histopathologic confirmation was done. 24% of 25 patients were leiomyoma, 20% polyp, 12% lipoma and the remainder were teratoma, carcinoid tumor, adenocarcinoma, polyp and leiomyosarcoma. EUS was successful in detecting all tumors. EUS diagnosis was consistent with histopathogical diagnosis in all patients with EUS findings as leiomyosarcoma (n=2) and polyp (n=6), in 46.2% of patients with EUS findings as leiomyoma, and in 50% of those with lipoma. CONCLUSIONS: EUS is an accurate means of evaluating and diagnosing submuocal lesions of the gastrointestinal tract. PMID- 15334316 TI - Is lower socio-economic status a risk factor for Helicobacter pylori infection in pregnant women with hyperemesis gravidarum? AB - BACKGROUND/AIMS: Lower socio-economic status was demonstrated to be a major risk factor for infection with H. pylori infection in the general population. The aim of the present study was to investigate any possible association between infection with H. pylori and hyperemesis gravidarum and between socio-economic status and H. pylori infection in pregnant women with hyperemesis gravidarum. METHODS: Fifty-six pregnant women with hyperemesis gravidarum and 90 control pregnant women were enrolled in the study. Forty-five (80.4%) subjects in the hyperemesis gravidarum group and 72 (80%) subjects in the control group were assessed as lower socio-economic status by questionnaire. Specific serum immunglobulin G for H. pylori was assayed by fluorescent enzyme immunoassay method for both groups. RESULTS: Serologically positive H. pylori infection was detected in 46 (82.1%) subjects of the hyperemesis gravidarum group and in 58 (64.4%) of the controls (p=0.024). With respect to the patients with lower socio economic status, 40 (88.9%) of the 45 patients with hyperemesis gravidarum and 49 (68.1%) of the 72 controls were positive for H. pylori (p=0.013). CONCLUSIONS: This study supports the studies suggesting an association between H. pylori and hyperemesis gravidarum. In addition, the lower socio-economic status may also be an important risk factor for infection with H. pylori in pregnant women with hyperemesis gravidarum. PMID- 15334317 TI - Comparison of ranitidine bismuth citrate, tetracycline and metronidazole with ranitidine bismuth citrate and azithromycin for the eradication of Helicobacter pylori in patients resistant to PPI based triple therapy. AB - BACKGROUND/AIMS: Helicobacter pylori is the most common infectious disease all over the world. Ten to twenty percent of the patients remain infected despite treatment with proton pump inhibitors (PPIs), amoxicillin and clarithromycin. We compared PPI, bismuth, tetracycline and metronidazole with ranitidine bismuth citrate, tetracycline and metronidazole in cases resistant to PPIs-based triple therapies. METHODS: The study included 52 patients who underwent a triple therapy with PPI, clarithromycin and amoxicillin for 14 days between September 2001 and December 2002, and were found to be resistant to the therapy. They were randomized to take ranitidine bismuth citrate (Rb) 400 mg twice a day, tetracycline (T) 1 g twice a day and metronidazole (M) 500 mg three times a day for 14 days (RbTM), or ranitidine bismuth citrate (Rb) 400 mg twice a day for 14 days and azithromycin (A) 500 mg once a day for 7 days (RbA). Four weeks after the treatment, endoscopies were repeated, and patients were assessed with respect to changes in symptoms. When H. pylori was negative on histological analysis and urease test, eradication was achieved. RESULTS: A total of 52 patients, 32 females and 20 males with a mean age of 49+/-12 years, were included in the study. Eradication was achieved in 15 (28%) out of 52 patients in total. There was a significant difference between RbA and RbTM groups (p=0.01). In fact, H. pylori was eradicated in 3 (12%) out of 25 patients in the RbA group, whereas it was eradicated in 12 (44.4%) out of 27 patients in the RbTM group. Symptom scores significantly improved in both groups after the treatment, though there was not a significant difference between the groups (p=0.705). CONCLUSIONS: Triple therapy including azithromycin does not seem to be a good choice in cases resistant to the first line therapies; however, a similarly lower rate of eradication was achieved with the quadruple therapy proposed. Therefore, different treatment schemes should be applied in resistant patients, and further studies are needed as well. PMID- 15334318 TI - Hepatic lymphoma metastasis presenting with severe acute liver failure: a rare case. AB - Hepatic lymphoma metastasis is rare, and should always be considered in the differential diagnosis of hepatic malignancy. A 52-year-old man presented with a four-day history of fever, fatigue, yellowish skin and nausea. His past medical history was unremarkable. There was no history of alcohol intake or medications. His physical examination revealed generalized jaundice and hepatomegaly. His blood tests showed liver failure and coagulopathy. Abdominal ultrasonography illustrated hepatomegaly. A further work-up included bone marrow and liver biopsy. The pathology report was B-cell lymphoma. He was treated with chemotherapy, and his laboratory findings during follow-up showed steady improvement. In conclusion, lymphoma metastasis to liver can be a cause of liver dysfunction. A high index of suspicion is required for the diagnosis. We emphasize the importance of obtaining tissue sample in all patients with suspicious lesion in any organ to avoid missing the rare but curable pathologies. PMID- 15334319 TI - Axonal neuropathy and hearing loss associated with alpha interferon treatment in chronic hepatitis B: a case report. AB - The effect of interferon alpha in chronic viral hepatitis and common side effects are well known, but axonal polyneuropathy and hearing loss have been rarely reported. A 58-year-old woman was administered interferon alpha-2a (9 MU/3 times a week) and lamividin (100 mg daily) with the diagnosis of chronic hepatitis B. At the fifth month of the treatment gait disturbance and tinnitus developed. In her neurological examination tandem gait was ataxic on the right side. Cerebral magnetic resonance imaging performed to elucidate a probable cerebral pathology revealed nonspecific millimetric hyperintense lesions thought to be related with her hypertension anamnesis. Electroneuromyography demonstrated mild axonal polyneuropathy. The finding of pure-tone audiometry was sensorineural hearing loss in her left ear. Our diagnosis was axonal polyneuropathy and sensorineural type hearing loss as a side effect of interferon. In conclusion, the development of polyneuropathy and sensorineural hearing loss in the same patient may suggest autoimmunity as the cause of these side effects. PMID- 15334320 TI - A case of Budd-Chiari syndrome secondary to multiple thrombogenic conditions: a case report and review of literature. AB - This report describes a case of Budd-Chiari syndrome caused by latent polycythemia vera and factor V Leiden mutation. This syndrome usually occurs due to thrombosis of hepatic veins or membranous obstruction of inferior vena cava. The most common reasons for thrombosis are manifest polycythemia vera and the other prothrombotic conditions. Recently, latent polycythemia vera and factor V Leiden mutation have been reported in increasing frequency. In this report, we aimed to emphasize that all prothrombotic conditions must be evaluated while investigating the etiology of Budd-Chiari syndrome, including latent polycythemia vera and factor V Leiden mutation, and appropriate antithrombotic and surgical therapies must be performed without delay. PMID- 15334321 TI - Duodenal varices as an unusual cause of gastrointestinal bleeding due to portal hypertension: a case report. AB - Duodenal varices that develop in patients with portal hypertension rarely cause hemorrhage, but varix rupture is a serious and often fatal event. We report the case of a 38-year-old man with hepatic vein occlusion who was referred to our hospital for gastrointestinal hemorrhage of unknown origin. Upper gastrointestinal endoscopy revealed varices in the distal third of the duodenum. These varices were identified as the source of the bleeding. The patient was treated with endoscopic band ligation, and with coil embolization of a shunt between the superior mesenteric vein and the left renal vein. PMID- 15334322 TI - Epiphrenic diverticulum with abnormal manometry: a case report. AB - A 50-year-old man was admitted to our hospital with the chief complaints of vomiting and nocturnal cough for one year. His complaints were progressive and had worsened within the last two months. Chest X-ray revealed a right-sided paracardiac opacity. Computed tomography of the thorax showed a supradiaphragmatic mass lesion containing air and solid components. Barium esophagogram revealed a giant distal esophageal diverticulum and hiatal hernia. Stationary manometric examination of the esophagus showed non-specific esophageal motility disorder. The patient underwent a successful abdominal approach diverticulectomy operation and the postoperative course was uncomplicated. There is a high prevalence of esophageal motility abnormalities in patients with epiphrenic diverticula, though they are very rare, and, as in this case, may be difficult to classify. PMID- 15334323 TI - The missing hazelnut. AB - This report describes a case of esophageal perforation caused by a hazelnut which became stuck in the upper esophagus but was not detected. We outline the pitfalls in diagnosis, complications and treatment in the pediatric population. PMID- 15334324 TI - A case of antiphospholipid antibody syndrome with Budd-Chiari and colonic ulcers complicated with gastrointestinal hemorrhage. AB - The antiphospholipid antibody syndrome (APA) is characterized by an increased incidence of venous and arterial thrombosis. APA syndrome has some gastroenterological manifestations such as Budd-Chiari syndrome, hepatic infarction, esophageal necrosis, intestinal ischemia, pancreatitis and colonic ulceration. We report a 34-year-old man with APA syndrome complicated by hepatic venous thrombosis (Budd-Chiari) and colonic ulcers. The clinical and laboratory findings were compatible with APA syndrome that developed secondary to systemic lupus erythematosus. In order to initiate anticoagulant therapy, he was heparinized. Since lower gastrointestinal bleeding developed, heparin was discontinued and the patient was followed up with baby aspirin and steroids. This case report extends the gastroenterological manifestations of the APA syndrome to include colonic ulceration, which may outweigh the efficacy of initial anticoagulant therapy. PMID- 15334325 TI - Inflammatory myoglandular polyp: a rare cause of hematochezia. AB - Inflammatory myoglandular polyp is characterized by inflammatory granulation tissue in the lamina propria, proliferation of smooth muscle, and hyperplastic glands with variable cystic change. The etiology is obscure. Mucous diarrhea, tenesmus, and hematochezia are main symptoms. We hereby report an 80-year-old man with diagnosis of inflammatory myoglandular polyp. PMID- 15334326 TI - Acute abdomen caused by a ruptured spontaneously infected mesenteric cyst. AB - Mesenteric cysts are uncommon and usually asymptomatic abdominal tumors. Spontaneous infection is a very rare complication of these lesions. This report presents a case of spontaneously infected and perforated mesenteric cyst. Any incidental mesenteric cyst diagnosed during radiological examination should be removed completely, even if it has been asymptomatic, to prevent potential life threatening complications. PMID- 15334327 TI - [Pain management in children and in the elderly]. PMID- 15334328 TI - [Pain treatment in neonates, infants and children--is the current treatment sufficient?]. AB - Paediatric patients quite often have to undergo painful or stressful procedures, e. g. blood sampling, dressing of wounds or removal of a drainage. The key problem is to decide if a child has pain or if there are other reasons for crying. Establishing a high standard in an institution requires regular evaluation and documentation of pain scores. For many clinical situations, clear and functioning concepts exist - we just have to use them. Unanswered questions are the evaluation of pain in small children, the side-effects of opioids, surgery involving the airways and the risk-benefit-ratio of certain techniques. Pain therapy after tonsillectomy is still troublesome: relevant postoperative pain occurs. Local infiltration of the tonsillar bed has no pre-emptive effect and only a minimal impact on the postoperative pain. Management relies on opioids, steroids and non-opioids. Non-steroidal anti-inflammatory drugs should not be used because of an increased risk of bleeding. Promising data have been reported on COX-2-blockers, but experience in children is still limited. Pain management after circumcision is relatively easy to perform. A conduction block with a long-acting local anaesthetic combined with one dose of a non-steroidal anti-inflammatory drug provides sufficient analgesia in over (2/3) of patients. Today, penile block is the standard of care and complications only rarely occur. However, despite successful pain prevention, circumcision remains a stressful procedure for the small patients. Pain treatment per se is not sufficient to relieve all the suffering connected with surgery in children. The concept of balanced analgesia is successful under many circumstances, but continuous efforts are needed to improve the management for difficult situations, e. g. tonsillectomy. PMID- 15334329 TI - [Use of opioids in the elderly -- pharmacokinetic and pharmacodynamic considerations]. AB - Perioperative management of geriatric patients is becoming an important component in anaesthetic practice in the 21st century. This phenomenon is due to the fact that people aged 65 and over are the segment with the fastest growing population. Thus, it is estimated that by the year 2025 20 % of the population in the western hemisphere will be > 65 years of age. Currently, elderly patients comprise one third of all operations, and one out of two patients older than 65 years of age will undergo an operation in their lifetime. The dramatic change in demographics of surgical patients will have a tremendous impact on the use of anaesthetics. Older patients facing surgery can generally be expected to be a more complex case than their younger counterparts. They have more systemic diseases (e. g. cardiac, pulmonary, endocrine), and usually these diseases have advanced to more serious stages. These patients may suffer disability, both physical and mental, and may show differences in the pharmacokinetic as well as the pharmacodynamic of compounds such as opioids. While neuronal numbers, dendrites and synapses decline with age and the ventricular volume triples, cerebral circulation is similar to young adults, although there is a reduction in cerebral blood flow (CBF). This is because of the lower unit weight, lower CBF and CMRO (2), which are tightly coupled in aging where autoregulation is preserved. However, because of a decline in dopaminergic, serotonergic, cholinergic and GABAergic transmitters, anticholinergic compounds (atropine, scopolamine) as well as some anaesthetics such as ketamine, benzodiazepines or even propofol may produce delirium and/or an increase in efficacy when given together with opioids. Therefore it is mandatory to consider a pharmacologic interaction with a potentiation and/or an addition in effects of other drugs when judging the net action of opioids in the elderly. Physicians and nurses treating geriatric patients tend to have an unfounded level of fear of complications associated with treating perioperative pain. Although it is known that inadequate analgesia may delay recovery, the treatment of perioperative pain in the geriatric patient remains inadequate, even relative to younger patients. It is well established that there is increased responsiveness to the effects of opioids in the elderly. This may result in an increased risk of respiratory depression, while especially the elderly female patient demonstrates an increase in the duration of effects, but the risk of nausea is not augmented. Increased sensitivity of older patients to systemic opioids mostly involves pharmacokinetic factors such as a higher proportion of unbound and active substances as well as changes in drug redistribution. Because of a 40 % reduction in stroke volume in the elderly, there is a protracted redistribution of opioids to the liver. This results in a prolonged metabolisation, a lesser inactivation over time followed by an increase in duration of effects, mainly impairment of respiration. To a much lesser extent, pharmacodynamic factors with an increased response at opioid receptor sites have to be considered. Although the mechanisms causing differences of opioid action in the elderly may be complex, the clinical implications are not. They include slow titration of opioids to allow for long circulation times, lower total doses because of increased sensitivity, and anticipation of a longer duration of action because of reduced clearance. Since elderly patients present multimorbidity, therapy of chronic pain has to be considered in the light of multidrug intake, which, due to interaction, results in marked side-effects, and a prolonged duration of action. Those opioids should be used which, due to their pharmacokinetic properties, have a reduced volume of distribution, present a low plasma protein binding and finally result in the formation of no pharmacologically active metabolites. PMID- 15334330 TI - [Effect of enteral naloxone on the incidence of gastritis and esophagitis in mechanically ventilated patients]. AB - BACKGROUND: Gastrtis and esophagitis are frequent and severe complications in intensive care patients, mainly caused by increased duodenogastral reflux. Opioids, commonly used in intensive care, are known to impair gastrointestinal (GI) motility which increases retrograd flow of gastric content. In a previous study, we showed that enteral administered naloxone reduces gastric reflux by selectively blocking GI opioid receptors. Therefore, in a subpopulation of these patients we studied the effect of enteral naloxone on the incidence of mucosal injury in opioid-treated, mechanically ventilated patients. METHODS: After IRB approval, mechanically ventilated, fentanyl-treated patients without gastrointestinal surgery or diseases were assigned to receive 8 mg naloxone or placebo four times daily via a gastric tube. Additional inclusion criteria were opioid treatment for at least three days and endoscopy of the upper GI tract. Frequency of gastritis and esophagitis was quantified according to the Savary Miller Score, and further parameter of GI motility (frequency of propulsive medication, amount of enteral feeding) were measured. RESULTS: In four of seventeen patients of the naloxone group (24 %) and 14 of 22 patients of the placebo group (64 %; p = 0.02), esophageal or gastral mucosal injuries were detected. In the naloxone group, gastral reflux as well as need for propulsive medication were significantly lower. Volume of enteral feeding showed an increasing trend in the second half of the study. CONCLUSION: Reduction of esophagogastral mucosal injury and reduced need for procinetic medication suggests an improvement of GI motility by enteral naloxone in fentanyl-treated, mechanically ventilated patients. PMID- 15334331 TI - [Evaluation of the disability of ventilated patients]. AB - OBJECTIVE: It is very important to investigate the patient's disability and pain. Interviews of intubated and tracheotomised patients were neglected because of inadaequate measuring methods. This is the first prospective study that evaluates the disability and pain of intubated and tracheotomised patients. METHODS: Disability, Hospital Anxiety and Depression Scale, Visuelle Analogue Scale, Glasgow Coma Scale and structured questions were used to investigate the disability and pain of ventilated patients. 26 patients of an interdisciplinary operative intensive care unit took part in the study. Sociodemographic parameters, ventilation, sedation and pain were evaluated. Doctors and nurses were asked to assess the patient's pain and disability. RESULTS: 17 intubated and 9 tracheotomised patients were included in the study. Mean intensity of pain was 30.3 (SD = 31.4), anxiety 40.8 (SD = 31.4), disability 30.0 (SD = 11.5) and disability caused by ventilation 61.9 (SD = 28.5). 46.2 % of the patients had a pathological subscale of anxiety and 50 % of depression in the Hospital Anxiety and Depression Scale. Nurses assessed the patient's disability and pain better than the doctors. CONCLUSION: A high disability has to be taken in account in the therapy of intubated and tracheotomised patients. PMID- 15334332 TI - [Rapidly lethal progression of a therapy-resistant status epilepticus]. AB - We present a case of death after first manifestation of generalised convulsive status epilepticus in a young man. A previously healthy 23-year-old man was admitted to our emergency department by ambulance service with approximately 20 min of generalised convulsive seizures. First line treatment in the emergency ward with benzodiazepines failed. The patient was cardiopulmonary stable until, after more than 30 min of status epilepticus, he developed tachycardia and became bradypnoeic. Intubation and ventilation was performed and anticonvulsive treatment was escalated with thiopental. Fifteen minutes later he developed ventricular fibrillation. CPR was started. The patient became asystolic after 90 min CPR following the ILCOR (International Liaison Committee on Resuscitation) Instructions. CPR was continued for another 30 min without success. The patient died after 120 min of maximal efforts. Autopsy and toxicology were performed, neuropathologic examination showed general brain edema and neuronal cell loss in purkinje cell layers of the cerebellum and olive knots which may be the consequence of generalised convulsive status epilepticus. We conclude: status epilepticus becomes refractory in approximately 30 % of cases. Until now, there are no randomised trials on the optimal treatment of refractory status epilepticus. Better treatment algorithms are urgently needed. PMID- 15334333 TI - [3rd International Symposium "Autologous transfusion--from euphoria to rational: practical handling from the scientific viewpoint" (Part I)]. PMID- 15334334 TI - [Outcome quality in terms of product safety in preoperative autologous blood donation]. AB - OBJECTIVE OF THE STUDY: The most common form of autologous blood transfusion is where blood is donated in anticipation of elective surgery. The aim of this study is to critically evaluate the product safety of autologous blood units. METHODS: The results are based on an analysis of the documented data and records of 22,630 cases with 21,553 patients and 49,650 autologous blood donations, blood processing and testing protocols, and reported transfusion reactions following autologous transfusions. RESULTS AND CONCLUSIONS: Autologous blood components are prepared individually, and an established system of quality assurance with controls of processes and products is particularly important to meet quality standards. The red cell content of autologous units cannot be standardized as in allogeneic components. Due to the differences between the laboratory screening programmes for patients and donors, the risk of transmission of virus infections is different from that of allogeneic blood components. The quality of stored autologous red cells is comparable to that of allogeneic products. Results of systematic sterility monitoring lead to the conclusion that under the conditions practised the risk of transfusion-associated sepsis is not higher to that of allogeneic products. Soluble biological response modifiers which accumulate during storage, are assumed to be the cause of reactions that occur in one of 4,500 autologous transfusions. Incorrect allocation and handling errors continue to be a serious problem in autologous transfusions. PMID- 15334335 TI - [Product quality in preoperative autologous blood donation--determinants of erythropoiesis]. PMID- 15334336 TI - [Quality assurance in blood salvage and variables affecting quality]. AB - Also intraoperative blood salvage (IBS) requires a system for quality management with controls of product an process quality. These can help early detection of malfunctions. For proper reaction and for improvement of quality a deep understanding of the process of blood salvage is necessary. This is supported by experimental testing of equipment, programs, and process variables, and by analysis of their effects on the process. The use of fresh whole blood and total protein as wash-out parameter in these tests is superior to outdated banked RBC and free haemoglobin. The process of aspiration turns out much less harmful than expected, when tested with fresh blood. Low wash volumes, fast washing rates, and half full bowls should be avoided. Plasma wash-out is improved by slower washing or by higher wash volumes, but the latter decrease RBC recovery. Such quality management helps to provide blood of excellent quality by IBS for optimal haemotherapy. PMID- 15334337 TI - Glycosylated hemoglobin in dogs: precision, stability, and diagnostic utility. AB - The precision and stability of the ion exchange chromatography assay for canine glycosylated hemoglobin (HbA(1)) were examined. The coefficient of variation (CV) of within-run replicate assays was 1.3 to 2.6%; the CV of between-run duplicate assays was 3.1%. The mean HbA(1) content in 44 healthy dogs was 7.1% (SD = 1.1%, range = 5.1-9.7%). Paired aliquots of 12 blood samples were stored at 4 degrees C and 25 degrees C, and HbA(1) was measured on the day of collection and at 3, 5, and 7 days after collection. In the blood stored at 4 degrees C, no significant increase in the HbA(1) content was seen. No significant increase in HbA(1) content was found in the blood stored at 25 degrees C after 3 days, but dramatic increases were observed after 5 and 7 days of storage. No significant difference was observed in the HbA1 content in heart blood collected 18 hours after death from 9 dogs kept at 25 degrees C. The HbA(1) content was measured in 10 hospitalized diabetic dogs. Five of the dogs had received no insulin and all 5 had elevated HbA(1) values. The other 5 dogs had received insulin for 1 to 9 months; 2 of the 5 had increased HbA(1) content. The HbA(1) content was determined periodically for 9 months in one diabetic dog and it declined from 14% to 8.2%. PMID- 15334338 TI - Mastocytemia in dogs with acute inflammatory diseases. AB - Nineteen dogs were identified that had mastocytemia (mast cells in venous blood samples) not associated with mast cell neoplasia. The first 10 dogs were identified by examination of blood films from dogs with suspected parvovirus enteritis (8), fibrinous pericarditis and pleuritis secondary to thoracic lacerations (1), and renal insufficiency of unknown cause (1). Because of the apparent association with acute enteritis, blood films from 52 suspected canine parvovirus cases were examined retrospectively and 8 mastocytemic dogs were found. An additional 52 canine blood films were randomly selected from the same retrospective time period and 1 mastocytemic dog was found that had pneumothorax, pelvic fractures, and hemorrhagic septic abdominal effusion secondary to renal hemorrhage and traumatized intestines. All mastocytemic dogs had acute inflammatory leukograms the day that mast cells were first detected: neutropenia with toxic neutrophils (4), neutropenia with a left shift (8), total neutrophil count within reference interval but with a left shift (5), or neutrophilia with a left shift (2). All dogs except the renal insufficiency case had circulating toxic neutrophils. Five dogs were mastocytemic on more than 1 day. The pathogenesis of the mastocytemia associated with the acute inflammatory disease was not determined. PMID- 15334339 TI - An enzyme immunoassay to measure canine circulating fibronectin. AB - A competitive enzyme immunoassay has been used to detect and quantitate fibronectin in canine plasma. In this test, purified fibronectin, bound to microtiter plates, competes with plasma fibronectin for the conjugated antibody, rabbit-anticanine, fibronectin-horseradish peroxidase. The assay could detect fibronectin in purified standards from 58 ng/ml to 580 microgram/ml. The range of 1-100 microgram/ml was linear for plasma samples diluted 1:10, allowing samples with fibronectin concentrations from 10-1000 microgram/ml to be easily measured by this method. The mean normal fibronectin concentration of 132 dogs, by this method, was determined to be 320 +/- 74 microgram/ml. PMID- 15334340 TI - Through the use of laboratory methods. PMID- 15334342 TI - Effects of I.M. injections on serum creatine phosphokinase (CPK) values in dogs. AB - Intramuscular injections of compounds of low irritancy (bicillin, valium, saline) were found to cause significant elevations in serum CPK in dogs. This route of treatment must therefore be avoided in clinical conditions where CPK values have diagnostic or prognostic importance. PMID- 15334341 TI - Plasma ammonia concentration in dogs using ion-exchange method of analysis. AB - Laboratory methods for measuring plasma ammonia concentration were reviewed. The ion-exchange method of analysis was evaluated using canine plasma from 15 clinically normal dogs and five dogs with portosystemic shunts. Mean and standard deviation was 40.3 +/- 8.1 microgram NH(3)/dl in normal dogs, and values ranged from 125 to 200 microgram NH(3)/dl in dogs with portosystemic shunts. Handling of blood samples properly for accurate results was found to require pre-chilled collection tubes, chilling the samples in an ice bath, immediate separation of plasma from red blood cells and immediate analysis for ammonia concentration. The ion exchange method was determined to be sufficiently accurate for clinical use and is suggested for any veterinary clinical pathology laboratory. PMID- 15334343 TI - A computerized retrospective study of liver enzyme concentrations in dogs. PMID- 15334344 TI - A technique for the electrophoretic separation of isoamylases. PMID- 15334345 TI - Evaluation of variation in serum globulin concentrations in dairy cattle. AB - BACKGROUND: Several factors may influence the concentration of serum globulins in healthy cows and thereby affect clinical interpretation; however, few studies have addressed sources of variation in globulin values. OBJECTIVES: The purpose of this study was to compare colorimetry-based total serum globulin values with electrophoretically-determined serum globulin fractions and with IgG concentration, and to evaluate diurnal and long-term physiological variation and the effects of lactation and venipuncture site on serum globulin concentrations in Holstein dairy cattle. METHODS: Serum total globulin and albumin concentrations were analyzed by colorimetry and electrophoresis in 86 lactating cows; IgG concentrations were determined by radial immunodiffusion in 41 dry and 34 lactating cows. Serum globulins were analyzed hourly for 24 hours in 8 lactating cows and weekly for 15 weeks in 6 additional cows. Globulin concentrations were compared in samples obtained from jugular and coccygeal venipuncture sites in 4 cows. Results were analyzed using parametric statistical tests. RESULTS: Colorimetry-based total serum globulin concentrations correlated well with gamma-globulin fractions (r2 = 0.87) and IgG concentrations (r2 = 0.91). Diurnal variation of total serum globulins concentration was significant (P =.01); however, globulins did not vary significantly over a 15-week period. Mean serum globulins concentration in samples obtained from the jugular vein was 2.35 g/L higher than that in samples obtained by coccygeal venipuncture (P <.0001). CONCLUSIONS: The colorimetric method used widely in routine laboratory analyses remains a useful test for globulins determination in dairy cattle. However, time of sampling and venipuncture site should be considered in the interpretation of serum globulins on serial or interindividual specimens. PMID- 15334346 TI - Inaccuracy of routine creatinine measurement in canine urine. AB - BACKGROUND: Urine creatinine concentration often is used in ratios such as urine protein:creatinine to compensate for dilution or concentration of spot urine samples. OBJECTIVE: The purpose of this study was to compare the accuracy of different techniques of urine creatinine measurement currently available for veterinary practitioners. METHODS: In 104 samples of canine urine diluted 1:20 with distilled water, creatinine concentration was measured using a kinetic Jaffe reaction assay, and an enzymatic technique on an automatic analyzer (Elimat) and 3 benchtop analyzers (Reflovet, Scil; Vitros DT2, Ortho-Clinical Diagnostics; Vettest 8008, IDEXX) used in veterinary practice. RESULTS: The Jaffe and enzymatic techniques on the Elimat were not significantly different, and their inaccuracy tested with human control urines was <5%. The benchtop analyzers underestimated creatinine concentration, especially at concentrations >2000 mg/L. Inaccuracy was higher with multilayer slide technology systems (Vitros and Vettest) than with the Reflovet system. Results were approximately 25% and 2% lower, respectively, than with the Elimat at urine creatinine concentrations about 2000 mg/L. CONCLUSION: Inaccuracy in urine creatinine measurements using benchtop analyzers should be taken into account when defining decision thresholds, which should be corrected according to the method used to avoid misinterpretations. PMID- 15334347 TI - Differential leukocyte counts determined in chicken blood using the Cell-Dyn 3500. AB - BACKGROUND: Automated hematology instruments commonly are used for mammalian blood analysis, but there is a lack of accurate automated methods available for avian leukocyte analysis. OBJECTIVE: The aim of this study was to validate differential leukocyte counts in blood from chickens using the Cell-Dyn 3500 hematology system and avian-specific software. METHODS: Blood samples were collected in lithium-heparin tubes from 2 groups (n = 84 and n = 139) of laying hens. Manual 200-cell differential counts were done on routinely-stained blood smears, and manual total granulocyte counts (heterophils and eosinophils) were done using an eosinophil stain in a counting chamber. Automated differential counts were done using VET 2.3, a research and development version of avian specific software for the Cell-Dyn 3500. Results were analyzed using Pearson's correlation and difference plots. RESULTS: Automated granulocyte counts from the Cell-Dyn were in good agreement with manual granulocyte counts (r = 0.93 and 0.80 for the 2 study groups). No correlation was found between automated and manual lymphocyte counts. Correlation coefficients for monocyte counts were 0.70 and 0.43. CONCLUSION: Automated leukocyte results from the Cell-Dyn using VET 2.3 software were not fully accurate. Total granulocyte counts may be of clinical usefulness, but results obtained for other parameters were unreliable. PMID- 15334348 TI - Internal quality control of a turbidimetric immunoassay for canine serum C reactive protein based on pooled patient samples. AB - BACKGROUND: Optimized internal quality control (IQC) procedures are important to ensure that only results without medically important errors are used for medical decision-making and to ensure that unnecessary rejection of valid analytical runs is avoided. Additionally, estimates of the analytical performance can be derived from IQC data. In the absence of available species-specific standards of a compound, the use of alternative control materials based on patient samples is a possibility, although investigations on the suitability of this approach are needed. OBJECTIVES: The objective of the study was to plan and implement a simple IQC procedure with control material based on pooled canine serum samples for a turbidimetric immunoassay (TIA) for the determination of human C-reactive protein (CRP) that recently was validated for the determination of canine serum CRP, and to assess the clinical analytical performance of the assay. METHODS: Proposed guidelines for the planning and implementation of IQC procedures were followed by using 2 control materials. Quality requirements of the assay were defined objectively by means of available data on biological variation, and goals for IQC performance were defined according to recommendations (probability of error detection [P(ed)] >.90 and of false rejection [P(fr)] <.05). Analytical performance was evaluated by means of medical decision charts. RESULTS: The control rule of 1(2.5s) (ie, rejection of the analytical run if at least 1 of 2 control materials deviates from the mean by more than 2.5 SD) fulfilled the criteria of predicted IQC performance (P(ed) =.94-1.00, P(fr) =.03). The IQC method was successfully implemented over a 14-week period. The observed coefficient of variation in the period of monitoring was 3.8% (low) and 2.9% (high), which equals excellent analytical performance. CONCLUSIONS: It was possible to plan and implement a simple IQC procedure for the CRP-TIA with control materials based on canine serum samples that fulfilled the criteria of high error detection and low false rejection of valid analytical runs. The assay showed excellent long-term analytical performance over a 14-week period. PMID- 15334349 TI - Detection of neoplastic lymphocytes in peripheral blood of dogs with lymphoma by polymerase chain reaction for antigen receptor gene rearrangement. AB - BACKGROUND: Uniquely rearranged immunoglobulin and T-cell receptor gene sequences can be amplified and electrophoretically separated by size to detect a clonal population of lymphocytes. OBJECTIVE: The purpose of this study was to determine whether the polymerase chain reaction (PCR) detects neoplastic (clonal) lymphocytes more frequently than do microscopic methods. METHODS: We identified neoplastic lymphocytes in peripheral blood by both routine and standardized microscopic examination of blood smears and by PCR amplification of blood-derived DNA and compared the 3 methods for frequency of detection of leukemic involvement. For standardized microscopic examination (200 leukocytes counted on Wright-Giemsa-stained blood smears), samples were categorized as negative (1% prolymphocytes, no lymphoblasts), or positive (>/=1 lymphoblast). A PCR-amplified sample was positive if 1 or 2 discrete bands were seen on the gel, or negative if no bands, a smear, or a faint ladder was seen. RESULTS: Using PCR, neoplastic lymphocytes were detected in peripheral blood 2.5 times more frequently than with routine or standardized microscopic evaluation. Eighty-three percent of samples negative by microscopy were positive by PCR. CONCLUSION: PCR is more sensitive than microscopy for the detection of clonal lymphocytes in peripheral blood. The results of this study also suggest that neoplastic lymphocytes circulate in peripheral blood at a higher frequency than previously reported. PCR may be useful for detecting or phenotyping lymphoma, monitoring response to therapy, identifying recurrence, and screening breeds at risk. PMID- 15334350 TI - Evaluation of the cytologic diagnosis of canine prostatic disorders. AB - BACKGROUND: Canine prostatic disease is commonly investigated using cytologic techniques, especially now that ultrasound-guided fine needle cell aspiration (US FNA) is widely available. Few studies, however, have evaluated the diagnostic accuracy of prostatic cytology. OBJECTIVE: The purpose of this study was to evaluate the usefulness of cytologic investigation of prostatic disease using US FNA and other methods in comparison with histopathologic diagnosis. METHODS: Cytologic and histopathologic specimens of prostate or paraprostatic tissue from 25 adult dogs were retrospectively evaluated. Cytologic samples were obtained by US-FNA, prostatic massage, or direct impression smears or aspirates of tissue at surgery. Histopathologic sections were obtained from tissue collected by biopsy or at necropsy. RESULTS: Cytologic diagnoses were categorized as nondiagnostic (n = 2); cyst (n = 1); squamous metaplasia (n = 2); inflammation (n = 4); benign prostatic hyperplasia (BPH; n = 5); inflammation and BPH (n = 3); inflammation, BPH, and neoplasia (n = 1); inflammation and neoplasia (n = 3); and neoplasia (n = 4). Cytologic diagnoses agreed with final histologic diagnoses in 20 of the 25 cases (80%). Of those samples collected by US-FNA, 75% were concordant. Four samples obtained by US-FNA and 1 sample obtained by prostatic massage and wash had discordant results. CONCLUSIONS: The results of this study suggest strong agreement between cytologic and histopathologic diagnoses for prostatic conditions. Discordance in results obtained by US-FNA usually was the result of the pathologic process rather than a failure to obtain an appropriate sample. PMID- 15334351 TI - Comparison of amylase and lipase activities in serum and plasma of dogs. AB - BACKGROUND: Amylase and lipase activities are most often determined in serum, although heparinized plasma is more convenient to obtain and is used for many routine biochemical analyses. OBJECTIVE: The purpose of this study was to compare amylase and lipase activities in serum and plasma of dogs and to determine whether either specimen type is acceptable for analysis. METHODS: Serum and heparinized plasma were obtained from 101 randomly selected dogs and analyzed in parallel for alpha-amylase and lipase. Results were compared using Passing Bablock regression, Bland-Altman difference plots, and correlation analysis. RESULTS: There was a high correlation between the results obtained from serum and those from plasma. Regressions (with 95% confidence intervals in parentheses) were as follows: lipase(plasma) = 0.984 (0.976/0.995) Chi lipase(serum) - 0.9 (2.9/0.7) (r =.999); a-amylase(plasma) = 1.003 (0.977/1.032) Chi alpha amylase(serum) - 1.9 ( 20.7/23.3) (r =.991). Mean differences (serum - plasma) were 8 U/L and 4 U/L for lipase and alpha-amylase, respectively. Classification of results as normal or abnormal did not differ according to specimen type. CONCLUSION: In dogs, lipase and alpha-amylase activities can be determined with the same level of accuracy in serum and in heparinized plasma. PMID- 15334352 TI - Vertebral telangiectatic osteosarcoma in a dog. AB - Telangiectatic osteosarcoma (TOS) affecting the seventh cervical vertebra (C7) was diagnosed in a 4-year-old male dog with signs of locomotor dysfunction. Bone lysis and an extradural-extramedullary mass were observed in radiographs and occipital myelograms. The diagnosis was confirmed by histopathologic and immunohistochemical examination of the mass. The tumor was composed of large blood-filled cavities lined by anaplastic stromal cells (malignant osteoblasts). Around the cavities were characteristic tumor giant cells ("bizarre cells"). Immunohistochemically the tumor cells were positive for proliferating cell nuclear antigen. The lining cells of the cysts were negative for von Willebrand factor. The histologic findings in this case of TOS involving C7 were similar to those seen in other cases of TOS in dogs and in people. Immunohistochemistry was a useful tool for assessing malignancy and for ruling out other differential diagnoses. PMID- 15334353 TI - Characterization of lymphocyte populations by flow cytometry in a calf with sporadic juvenile lymphoma. AB - A 2-month-old Angus heifer was presented to Washington State University Veterinary Teaching Hospital for generalized peripheral lymphadenopathy and mild anorexia. Results of a CBC revealed marked lymphocytosis consisting primarily of large, atypical lymphocytes with cleaved, reniform nuclei. A presumptive diagnosis of sporadic juvenile lymphoma was made. To confirm these findings, a prescapular lymph node was submitted for histopathology, immunochemistry, and flow cytometric analysis. Peripheral blood also was analyzed by flow cytometry. Phenotypic characterization of lymph node and peripheral blood cell populations verified the calf had the B-cell form of sporadic juvenile lymphoma. The results of this investigation expand the phenotypic characterization of neoplastic lymphocytes in sporadic juvenile lymphoma. PMID- 15334354 TI - Calcospherite-like bodies and caseous necrosis in tracheal mucus from a dog with tuberculosis. AB - A 3-year-old Wirehaired Fox Terrier was presented to the University Veterinary Hospital, University College Dublin, for evaluation of chronic cough of 8-months duration. Bronchoscopy showed a severely dilated collapsed left principal bronchus filled with highly viscous white mucus. Cytologically, globular lipid like material and round concentrically laminated crystalline structures were evident within the proteinaceous mucus. These findings resembled the calcospherites and granular caseous debris often observed in human tuberculous patients. A Ziehl-Neelsen-stained cytocentrifuged preparation of material obtained by bronchoalveolar lavage revealed a few acid-fast rods within macrophages, suggestive of tuberculosis. At necropsy, granulomas with caseous necrosis were present in the lung parenchyma, bronchial and mediastinal lymph nodes, liver, pancreas, and mesentery. Granulomas were adherent to both kidney capsules and to the diaphragm. Histologically, there was evidence of mild calcification within caseous granulomas, which was confirmed by von Kossa's stain. Using Ziehl-Neelsen stain, acid-fast rods were identified within granulomas; bacterial culture was positive for Mycobacterium bovis. The cytologic findings in this case have not been reported previously in dogs and demonstrate a possible correlation between tuberculosis and calcospherite-like bodies with caseous, globular material in bronchial mucus, similar to that described in human patients. PMID- 15334355 TI - Parasitemia in a neonatal bison calf. AB - A 3-day old female bison calf (Bison bison) was presented in lateral recumbency to the Universite de Montreal Veterinary Teaching Hospital. The animal was severely depressed and dehydrated (10%) and died a few hours after admission. Prior to death, blood samples were obtained for CBC, clinical chemistry, and serology tests. Abnormal CBC findings included thrombocytopenia, lymphocytosis, mild monocytosis, and a toxic left shift. Abnormal serum clinical chemistry findings included marked azotemia, hyperphosphatemia, hypernatremia, hyperalbuminemia, hypoglobulinemia, and low gamma-glutamyltransferase activity. Serologic test results for bovine leukosis virus and bovine viral diarrhea virus were negative. Blood smear examination revealed numerous elongated organisms that were tapered at both ends and characterized by an undulating membrane and a long flagellum. The organisms ranged in length from 35 to 40 micro meter, excluding the flagellum, and were identified as Trypanosoma theileri. Postmortem examination revealed that the animal suffered from concurrent mycotic abomasitis and colisepticemia. PMID- 15334356 TI - Mycobacterium tuberculosis infection in a dog from Africa. AB - A 4-year-old male Boxer dog with a history of vomiting, diarrhea, and weight loss moved from West Africa to Lyon, France, where it was further evaluated. Radiographs revealed pleural effusion and enlargement of tracheobronchial lymph nodes and liver. Cytologic examination of the pleural effusion and a fine needle aspirate specimen of the liver showed mixed mononuclear inflammation with nonstaining rod structures within epithelioid histiocytes. At necropsy, the main gross pathologic findings were exudative pleuritis, nodular hepatitis, and infarcts and caseous nodules in the kidneys. The main histologic lesions were granulomatous hepatitis, granulomatous pneumonia, fibrinous leukocytic pleuritis, necrotic and fibro-calcified granulomatous lymphadenitis, and granulomatous nephritis. A Ziehl-Neelsen stain applied to both cytologic and histologic samples was positive for acid-fast bacilli. Bacterial culture of the pleural fluid was positive for Mycobacterium tuberculosis. Cytology is a valuable tool in the diagnosis of this important zoonotic disease. PMID- 15334357 TI - Antithrombin III (ATIII) activity in plasmas from normal and diseased horses, and in normal canine, bovine and human plasmas. AB - Plasma Antithrombin III (ATIII) activity was quantitated in 24 clinically normal Standardbred/Thoroughbred horses using a clotting time technique. ATIII activity ranged from 80 to 106% of the pooled reference standard plasma, with a mean of 94%. Horses presenting with impaction or spasmotic colic (n=17) had normal plasma ATIII activity, while 15 horses presenting with acute diarrhea/colitis had significantly lower plasma ATIII activity with a mean of only 74% of the reference plasma. Seven horses presenting with liver disease had significantly higher plasma ATIII activity with a range of 127 to 177% of the pooled reference plasma. Fifty-seven equine plasmas were retested using a rapid chromogenic substrate technique for quantitating plasma ATIII activity. A good correlation (r =+0.83) existed between clotting time and chromogenic determinations of ATIII. Pooled normal canine, human and bovine plasmas had only 65, 62 and 79%, respectively, of the ATIII activity of the equine reference plasma. PMID- 15334358 TI - What is your diagnosis? PMID- 15334359 TI - Dyserythropoeisis, sideroblasts/siderocytes and hemoglobin crystallization in a dog. AB - Dyserythropoiesis characterized by enhanced intramedullary destruction, pathologic sideroblasts and siderocytes, and hemoglobin crystallization was detected in a female Cocker Spaniel presented for poor exercise tolerance. Examination of peripheral blood revealed intraerythrocytic crystals, granulation of erythrocytes, nucleated erythroid cells, reticulocytosis and marked variation in erythrocyte morphology in the absence of anemia. Bone marrow examination revealed sideroblasts, a low M:E ratio and evidence of enhanced intramedullary destruction of erythroid cells. Electron microscopy of peripheral blood and bone marrow confirmed pathologic mitochondrial iron accumulation in erythroid cells and the presence of intraerythrocytic hemoglobin crystals. A cause for the hematologic changes was not identified. After the animal became clinically normal, siderocytes disappeared from peripheral blood but intraerythrocytic crystals and reticulocytosis persisted. PMID- 15334360 TI - Free radicals: relation to tissue damage - a review. AB - Free radicals are any molecules having an odd number of electrons. These molecules are highly reactive and can be generated as byproducts of normal metabolism as well as by exposure to a number of environmental factors including drugs, radiation and air pollutants. Due to the ubiquity of molecular oxygen, the oxygen metabolites superoxide anion, hydrogen peroxide, and the hydroxyl radical are frequently involved in both beneficial and detrimental free radical reactions. Intracellular enzymes and radical scavengers help to protect against tissue damage by these reactive metabolites. The extent of free radical damage to tissue depends on the nature of the radical produced and its site of generation. PMID- 15334361 TI - Comparison between serum haptoglobin and alpha-2-globulin concentrations in dogs. PMID- 15334362 TI - Potential role of serum inhibitors of erythropoiesis in the anemia associated with infection, renal disease and malignancy in the dog. AB - Serum from dogs with anemia secondary to infection, malignancy and chronic renal disease was tested for the capacity to inhibit in vitro erythropoiesis. Serum inhibition was found in seven of 10 dogs with infectious diseases, four of 10 dogs with malignant diseases, and two of 10 dogs with chronic renal failure. The results were reproducible with at least three different marrow donor dogs. Serum inhibition did not correlate with decreased packed cell volumes. However, serum inhibition occurred in the two dogs with greatest azotemia suggesting that inhibitors of erythropoiesis are present only in advanced stages of renal failure. PMID- 15334363 TI - Mean platelet volume artifacts: the effect of anticoagulants and temperature on canine platelets. AB - Improper handling of specimens results in artifactually high Mean Platelet Volume (MPV) measurements limiting their usefulness as a clinical tool. MPV measurement and Scanning Electron Microscopy (SEM) were performed on split specimens collected from normal dogs using two anticoagulants and two temperatures over a period of 4 hours. Platelets exposed to EDTA and maintained at 4 degrees C (39.2 degrees F) exhibited the highest artifactual increase in MPV, while those exposed to citrate and maintained at 37 degrees C (98.6 degrees F) exhibited minimal change. The increase in MPV was accompanied by platelet shape change from a smooth disc to an irregular sphere with filopodia. It is recommended that citrated specimens maintained at 37 degrees C be used in all MPV measurements. PMID- 15334364 TI - What is your diagnosis? PMID- 15334365 TI - A review of feline serum protein electrophoresis. AB - A review of the current literature available on feline serum proteins is presented. Early studies concentrated on comparative aspects of species variations in the electrophoretic pattern. The feline electrophoretogram was divided into five basic regions: albumin, alpha-1, alpha-2, beta and gamma. Different subdivisions of these areas were recognized depending on the support medium used. Current papers have compared the relative migration distances of each globulin peak to the migration of albumin. This "Rf" value enables reliable peak identification. To date, no data exists identifying the individual proteins responsible for the peaks in the alpha and beta regions. The only feline globulin to be studied is haptolobin; however its precise location on the electrophoretic strip was not identified. PMID- 15334366 TI - Effects of weight loss in obese subjects with normal fasting plasma glucose or impaired glucose tolerance on insulin release and insulin resistance according to a minimal model analysis. AB - We investigated effects of weight loss from diet and exercise regimen in obese subjects with normal fasting plasma glucose or impaired glucose tolerance (IGT) on insulin release capacity and insulin sensitivity. Eight subjects were recruited among visceral obesity patients (4 men, 4 women; age range, 24 to 57 years; body mass index [BMI], 32.8 to 60.3 kg/m(2)). All were admitted to Chiba University Hospital for 2 weeks, were treated with a tapering 5,023 to 2,930 kJ diet, and were given exercise equivalent to 628 kJ/d. For assessments, we used a combination of C-peptide secretion rate determination and minimal model analysis as previously reported. BMI and visceral fat area (V) significantly decreased (BMI on initiation v after intervention, 43.0 +/- 3.2 v 40.3 +/- 3.1 kg/m(2), P <.05; V, 224 +/- 22 v 188 +/- 22 cm(2); P <.05). Fasting immunoreactive insulin (F-IRI) and leptin concentrations decreased significantly. Capacity for insulin release in response to glucose increased in all subjects (first-phase insulin secretion [CS1], 4.66 +/- 4.05 v 6.81 +/- 4.57 ng/mL/5 min, P <.05), but the insulin sensitivity index (S(i)) did not change significantly. These data suggest that weight reduction early in development of type 2 diabetes can oppose progression of diabetes by improving capacity for insulin release. PMID- 15334367 TI - Elevated serum parathyroid hormone concentration in eucalcemic patients after parathyroidectomy for primary hyperparathyroidism and its relationship to vitamin D profile. AB - Elevation of serum parathyroid hormone (PTH) level in eucalcemic patients after parathyroidectomy for primary hyperparathyroidism has been described in up to 40% of patients, but little is known about its etiology or clinical significance. To better understand the cause of this phenomenon, we studied 49 patients without renal dysfunction or osteomalacia who underwent parathyroidectomy for primary hyperparathyroidism. Patients were categorized into 2 groups based on their serum PTH and calcium levels after parathyroidectomy: (1) elevated PTH with eucalcemia (n = 21), (2) normal PTH with eucalcemia (n = 28). Elevation of serum PTH with eucalcemia after parathyroidectomy occurred in 43% of patients. Patients in group 1 had significantly higher preoperative and postoperative mean serum PTH levels and significantly lower postoperative serum levels of 1,25(OH)(2)D(3), 1,25(OH)(2)D(3)/25(OH)D(3) ratio, and 1,25(OH)(2)D(3)/PTH ratio compared with patients in group 2. Serum PTH in group 1 patients normalized as early as 3 months, but remained elevated in some patients for more than 4 years, and was not associated with development of recurrent hypercalcemia. Normalization of serum PTH in group 1 patients was associated with significant increase in 1,25(OH)(2)D(3) and 1,25(OH)(2)D(3)/PTH ratio. Our data suggest that elevation of serum PTH in eucalcemic patients after parathyroidectomy is a frequently reversible state of resistance of the kidneys to PTH-mediated 1-alpha hydroxylation of 25(OH)D(3) and does not signify subsequent recurrence of hyperparathyroidism. PMID- 15334368 TI - Correlation of oral glucose tolerance test-derived estimates of insulin sensitivity with insulin clamp measurements in an African-American cohort. AB - The purpose of this study was to determine which measures obtained from an oral glucose tolerance test (OGTT) are the best estimates of insulin sensitivity measured directly using the euglycemic hyperinsulinemic clamp procedure. Data were examined from a study conducted on 307 young adult African-American men and women. An OGTT with insulin measurements was conducted after a 12-hour overnight fast. The euglycemic hyperinsulinemic clamp was used to measure insulin stimulated glucose uptake (M) directly. Pearson's correlation analyses were performed to examine the relationship of OGTT-derived parameters with insulin sensitivity measured using the clamp. There were consistent statistically significant correlations between calculated estimates of insulin sensitivity (fasting insulin/fasting glucose, summed insulin/summed glucose, homeostasis model assessment [HOMA], Quantitative Insulin Sensitivity Check Index [QUICKI]) with insulin sensitivity measured by the insulin clamp (P <.001). The calculated estimates that correlated most strongly with clamp measured insulin sensitivity were QUICKI and the logarithm of summed insulin during the OGTT. These data indicate that fasting and OGTT-derived plasma insulin and glucose concentrations can be used to estimate insulin sensitivity in young adult African-Americans when it is not feasible to conduct the insulin clamp procedure. Calculated indices that include log transformation of plasma insulin concentration improve the estimation of insulin sensitivity. PMID- 15334369 TI - Effects of atorvastatin on triglyceride-rich lipoproteins, low-density lipoprotein subclass, and C-reactive protein in hemodialysis patients. AB - Dyslipidemia is an important risk factor for cardiovascular disease in patients with chronic renal failure (CRF). We evaluated the safety and efficacy of atorvastatin in patients with dyslipidemia associated with CRF who were undergoing hemodialysis (HD). Thirty-five patients who were receiving HD were given atorvastatin (10 mg/d) for 3 months. Chylomicron (CM), light and dense very low-density lipoprotein (VLDL), intermediate-density lipoprotein (IDL), and light and dense low-density lipoprotein (LDL) were separated by ultracentrifugation. Apolipoprotein (apo) B was measured by electroimmunoassay. Mean LDL particle diameter was measured by gradient gel electrophoresis. Atorvastatin therapy reduced LDL-cholesterol (C) by 36% and remnant-like particle (RLP)-C by 58%. Atorvastatin significantly reduced apo B, apo CIII, and apo E in VLDL by 40% to 46% and IDL-apo B by 66%. Atorvastatin also significantly reduced cholesterol in CM, light VLDL, and dense VLDL without consistently affecting triglyceride (TG) in these lipoproteins. Atorvastatin similarly reduced both light and dense LDL apo B by 38%. LDL particle size in the HD patients significantly increased during atorvastatin treatment from 25.7 +/- 0.4 to 26.2 +/- 0.6 nm. High sensitive C reactive protein (HS-CRP) was halved by atorvastatin decreasing from 0.08 +/- 0.05 to 0.04 +/- 0.03 mg/dL. Atorvastatin treatment did not affect the creatinine kinase level, and no classical adverse effects were observed during the study. These results suggest that atorvastatin is safe and effective for the management of dyslipidemia in patients with CFR who are receiving HD, which may help to suppress the development of atherosclerosis. PMID- 15334370 TI - F2 isoprostane is already increased at the onset of type 1 diabetes mellitus: effect of glycemic control. AB - Much evidence has suggested that oxidative stress (OS) may play a role in the pathogenesis of diabetic complications. However, the relationship between hyperglycemia and OS is inconsistent in diabetic clinical studies. The aim of this study was to evaluate the effect of normalization of blood glucose levels on urinary 8-epi-prostaglandin F(2alpha) (8-epi-PGF(2alpha)) excretion at the onset of type 1 diabetes. We studied 14 type 1 diabetic patients (50% males; mean age, 24.3 +/- 4.9 years) and 14 control subjects matched by age and body mass index. A 24-hour urine collection was performed to determine 8-epi-PGF(2alpha) as an integrated index of OS production at baseline, before starting insulin therapy, and 16 weeks later. Insulin treatment induced a significant reduction in glycosylated hemoglobin (HbA(1c)) (from 11.5% to 5.4% P =.0001), triglycerides (from 1.0 to 0.8 mmol/L, P =.002), and an increase in high-density lipoprotein (HDL)-cholesterol levels (from 1.1 to 1.5 nmol/L, P =.01) at week 16. This improvement in metabolic control was associated with a statistically significant reduction in 8-epi-PGF(2alpha) values (from 92.0 +/- 41.5 to 66.9 +/- 28.9 pg/mg urinary reatinine excretion, P =.015), although compared with the control group, 8-epi-PGF(2alpha) values remained higher in diabetic patients (66.9 +/- 28.9 v 39.1 +/- 13.8 pg/mg creatinine, P =.004). Enhanced OS is present in early clinical phases of type 1 diabetes, and the amelioration in metabolic control is associated with improvement in this pathogenic pathway. PMID- 15334371 TI - Rosiglitazone treatment improves insulin regulation and dyslipidemia in type 2 diabetic cynomolgus monkeys. AB - Impairment of peroxisome proliferator-activated receptor-gamma (PPAR-gamma), a nuclear receptor that regulates genes involved in lipid and glucose metabolism, may contribute to the onset of metabolic disorders such as diabetes and the accompanying dyslipidemia. Fat-derived tumor necrosis factor alpha (TNF-alpha) and the acute-phase response protein, C-reactive protein (CRP), may also have a role in the development of obesity-related insulin resistance and type 2 diabetes mellitus. In this study, a group of 14 naturally occurring, insulin-requiring, type 2 diabetic cynomolgus monkeys were used to evaluate the effects of the PPAR gamma agonist, rosiglitazone, on glycemic and lipid parameters and serum levels of TNF-alpha and CRP. The animals were randomized into 2 groups of 7. One group was treated with 0.5 mg/kg rosiglitazone orally once a day for 7 weeks. Blood was collected for evaluation at baseline, at 2 and 7 weeks during the treatment period, and at 7 and 13 weeks after treatment. Daily insulin requirements were recorded during the entire study. Results showed daily exogenous insulin requirements were significantly reduced (P <.01) in those treated with rosiglitazone, while glycemic control was maintained. Plasma triglyceride concentrations were significantly lower (P <.01) whereas plasma cholesterol levels tended to be lower and high-density lipoprotein (HDL) concentrations tended to be higher after treatment. No significant differences were noted in TNF alpha and CRP serum levels during the treatment period. Body weights remained steady in both groups during the study. These results suggest overall improvement in insulin regulation and lipid profiles during treatment with rosiglitazone. PMID- 15334372 TI - Carbohydrate requirement and insulin concentration during moderate exercise in type 1 diabetic patients. AB - The lack in control of insulin release combined with an inadequate carbohydrate (CHO) ingestion accounts for the occurrence of frequent metabolic unbalances during exercise in type 1 diabetic patients. The aim of the study was to quantify, in these patients, the CHO requirement to prevent hypoglycemia during moderate exercise performed at different time intervals after morning subcutaneous insulin injection. Twelve type 1 diabetic patients and 12 well matched healthy subjects cycled 4 times for 1 hour at a constant workload. The rate of glucose oxidation was calculated continuously by indirect calorimetry throughout the exercise, while blood parameters were assessed periodically and orally given CHO were checked. CHO needed by the patients to prevent hypoglycemia decreased as the time elapsed from insulin administration increased, amounting to 0.63 +/- 0.30, 0.44 +/- 0.32, 0.28 +/- 0.24, and 0.14 +/- 0.18 g/kg after 1, 2.5, 4, and 5.5 hours, respectively. Total glucose requirement during moderate exercise (sum of alimentary and extracellular source) was correlated (r = 0.739, P <.001) to plasma insulin concentration, but not with fitness level. Time elapsed from last insulin dose is not a factor influencing the risk of hypoglycemia during exercise when a proportional, appropriate amount of CHO is ingested. PMID- 15334373 TI - Fat oxidation after acipimox-induced reduction in plasma nonesterified fatty acids during exercise at 0 degrees C and 20 degrees C. AB - The main aim of this study was to investigate if whole body fat oxidation, after acipimox administration, during submaximal exercise in the cold, is different from that at temperate environments. Seven healthy recreationally active male subjects cycled at 70% Vo(2peak) for 60 minutes; once at 0 degrees C and once at 20 degrees C. To exclude availability, and therefore oxidation of plasma-derived nonesterified fatty acids (NEFA), 90 minutes before each cycling bout, subjects ingested 250 mg of the antilipolytic drug, acipimox. Blood and expired gas measurements were obtained at rest, immediately before exercise, and at 15, 30, 45, and 60 minutes of exercise. In both trials, after the ingestion of acipimox, plasma NEFA concentrations fell dramatically and immediately before and during exercise were lower than 0.05 mmol. L(-1) in both trials. Pre-exercise and exercise values of glycerol, glucose, triacylglycerol (TG), and rectal temperature (T(re)) were not different between the 0 degrees C and 20 degrees C trials. During exercise at 0 degrees C, skin temperature (T(sk)) was significantly reduced from pre-exercise values (P <.05) and at all time points was significantly lower than during exercise at 20 degrees C. Muscle temperature did not differ between trials but in both trials was lower (P <.05) at 1 cm depth than at 3 cm and 2 cm. Gross energy expenditure of cycling (0 degrees C trial, 3.6 +/- 0.1 MJ; 20 degrees C trial, 3.6 +/- 0.1 MJ), the oxidation rates of carbohydrate (0 degrees C, 32.4 +/- 0.5 KJ. min(-1); 20 degrees C, 32.6 +/- 0.7 KJ. min(-1)) and fat (0 degrees C, 24.6 +/- 1.2 KJ. min(-1); 20 degrees C, 23.0 +/- 1.8 KJ. min(-1)), and the proportion of energy derived from fat (0 degrees C, 45 +/- 1 %; 20 degrees C, 40 +/- 4%) and carbohydrate (0 degrees C, 55 +/- 1%; 20 degrees C, 58 +/- 3%) were not different between the 2 trials. In conclusion, after acipimox administration, whole body fat oxidation during exercise, designed to avoid adjustment of core temperature or thermogenesis, is not different at 0 degrees C compared with 20 degrees C. This allows the inference that during submaximal exercise, cold has no effect on the utilization of intramuscular TG (IMTG). PMID- 15334374 TI - Lack of relationship between beta3-adrenergic receptor gene polymorphism and gestational diabetes mellitus in a Taiwanese population. AB - The Trp64Arg polymorphism of beta(3)-adrenergic receptor (ADRB3) has been reported to be associated with insulin resistance and gestational diabetes mellitus (GDM). The objective of this study is to investigate whether the ADRB3 Arg variant confers susceptibility to GDM in a Taiwanese population. A total of 299 pregnant women (mean +/- SD, 31.1 +/- 4.2 years) was recruited. Two-hour, 75 g oral glucose tolerance tests (OGTT) were conducted at 24 to 31 weeks gestation (28.3 +/- 1.6 weeks). Forty-one GDM subjects and 258 controls with normal glucose tolerance (NGT) level were genotyped for the ADRB3 Trp64Arg polymorphism. The genotype distribution and allele frequency of ADRB3 did not significantly differ between GDM and NGT subjects (9.8% v 14.5%). Body weight gain during pregnancy was not different between ADRB3 genotypes. However, the GDM subjects with the Arg64 variant had higher fasting (P =.04) and postload 120 minutes (P =.03) insulin levels as compared with the GDM subjects with the Trp64Trp allele. In all subjects, the women with the Arg64 allele (n = 76) had significantly higher level of insulin secretion (the ratio of Deltainsulin(60)/Deltaglucose(60)) during OGTT than those without (n = 223) (P =.03) despite similar plasma levels of glucose and insulin in both genotypes. Our results indicated that the ADRB3 Trp64Arg variant is not related to the development of GDM and has no effect on obesity during pregnancy in a Taiwanese population. However, ADRB3 polymorphism might be a possible determinant of insulin resistance. PMID- 15334375 TI - Difference between diabetic and nondiabetic smokers in the pituitary response to physical exercise. AB - The growth hormone (GH), cortisol, and arginine vasopressin (AVP) responses to bicycle ergometry (with increasing workload until exhaustion) were measured in 20 patients affected by insulin-dependent diabetes mellitus (IDDM) (10 habitual smokers and 10 nonsmokers) and 20 nondiabetic subjects (normal controls) (10 habitual smokers and 10 nonsmokers). Cardiorespiratory parameters such as heart rate, blood pressure, ventilation, frequency of breathing, tidal volume, oxygen consumption (Vo(2)), carbondioxide production (Vco(2)), and respiratory exchange ratio (R) were monitored before and during exercise tests. No significant differences between groups were observed; furthermore, there were no differences in circulating somatomedin-C (SM-C) and free fatty acids (FFA) levels between groups. Blood glucose levels were similar before exercise and followed a similar pattern during tests in diabetic smokers and nonsmokers. Basal GH, cortisol, and AVP levels were similar in diabetic smokers, diabetic nonsmokers, normal smokers, and normal nonsmokers. In all groups, exercise induced a significant increase in the serum concentrations of all examined hormones. Increments were significantly higher in diabetic than in nondiabetic groups. No significant differences were observed between diabetic smokers and nonsmokers for all examined hormones. AVP responses during tests were similar in normal smokers and nonsmokers. In contrast, exercise-induced GH and cortisol increments were significantly lower in normal smokers than in normal nonsmokers. These data support the hypothesis that in normal subjects habitual nicotine consumption may attenuate both GH and cortisol responses to a releasing stimulation, such as physical exercise. This phenomenon may represent an expression of adaptation of nicotinic neurotransmission to chronic stimulation. Furthermore, the data show that the effect induced by habitual smoking is absent in diabetics, probably because of diabetes-induced neuroendocrine alterations in the central nervous system. PMID- 15334376 TI - Codelivery of a tea extract prevents morbidity and mortality associated with oral vanadate therapy in streptozotocin-induced diabetic rats. AB - Oral administration of vanadate has a strong hypoglycemic effect but results in toxic side effects like life-threatening diarrhea. Tea is known to have potent antidiarrhea effects. We investigated the potential of suspending the vanadate in a tea decoction to reduce the diarrheatic action of vanadate. A concentrated extract of Lichee black tea was, therefore, added to sodium orthovanadate. Streptozotocin (STZ)-induced diabetic rats were orally gavaged with vanadate suspended in water or in the tea decoction, or with the tea extract alone. Blood glucose levels were assessed daily over 11 weeks with levels greater than 10 mmol/L warranting therapeutic intervention. Both the vanadate/water and vanadate/tea solutions acutely reduced blood glucose. The tea extract alone had no effect. The majority of vanadate/water-treated rats developed diarrhea and mortality rates approached 40%. Vanadate/tea-treated diabetic rats experienced no diarrhea or mortality and liver and kidney analyses (plasma ALT and creatinine, blood urea nitrogen [BUN], and urine-specific gravity) were normal. Animals treated with vanadate/tea retained blood glucose levels less than 10 mmol/L for an average of 24 consecutive days without subsequent treatments. Cataract formation was completely prevented. The mechanism of action of vanadate may have involved beta-cell stimulation because vanadate/tea-treated diabetic rats exhibited normal plasma insulin levels. In summary, because of its long-lasting effects, oral administration, and lack of side effects, vanadate/tea represents a potentially important alternative therapy for an insulin-deficient diabetic state. PMID- 15334377 TI - Association between the corrected QT intervals and combined intimal-medial thickness of the carotid artery in patients with type 2 diabetes. AB - The main purpose of this study was to determine whether cardiac autonomic neuropathy or coronary atherosclerosis is the more important factor affecting prolongation of the corrected QT interval (QTc) in patients with type 2 diabetes. We studied the association between QTc and the coefficient of variance of the heart rate variation (CV(RR)), which reflects cardiac autonomic neuropathy, and the combined intimal-medial thickness (IMT) of the common carotid artery, which reflects coronary atherosclerosis. In addition, we also investigated the relationship between the QTc and blood pressure, serum lipid concentrations, hemoglobin A(1C) (HbA(1C)) concentration, and duration of diabetes. We studied 75 patients with type 2 diabetes and 30 age-matched healthy individuals. The QT interval was measured in lead II of the electrocardiogram (ECG) and was corrected using Bazett's formula. Cardiac neuropathy was assessed by measuring CV(RR). Atherosclerosis was evaluated by measuring the combined IMT of the common carotid artery using B-mode ultrasonography. The QTc in patients with type 2 diabetes was significantly longer than in healthy individuals (P <.0001). The QTc more closely correlated with the IMT of the carotid artery (r = 0.7206, P <.0001), compared with CV(RR) (r = -0.3188, P =.0053), although both were statistically significant. The QTc also correlated positively with the systolic (SBP) and diastolic blood pressure (DBP) (r = 0.4371, P <.0001, r = 0.3632, P =.0014, respectively). Based on stepwise regression analysis with the QTc interval as the dependent variable, the IMT of the carotid artery had the most significant association with the QTc (beta = 0.6882, P =.0004). In conclusion, QTc prolongation in the setting of diabetes might be caused primarily by coronary atherosclerosis rather than by cardiac autonomic neuropathy. PMID- 15334378 TI - Long-Term exposure of INS-1 cells to cis and trans fatty acids influences insulin release and fatty acid oxidation differentially. AB - The importance of elevated levels of fatty acids in the pathogenesis of the deteriorated beta-cell function present in type 2 diabetes has been established. Long-term exposure of the beta-cell to high levels of fatty acids causes enhanced insulin secretion at low glucose (basal insulin release), while glucose stimulated insulin secretion (GSIS) is decreased or unchanged. We have previously demonstrated that the spatial configuration of fatty acids (cis and trans isomers) is of importance for the acute impact on the beta-cell function. In this study we aimed to elucidate whether the spatial configuration also influenced beta-cell function after long-term exposure. Thus, we compared the effect of 3 days culture of INS-1 cells with cis (cis C 18:1-11) and trans vaccenic acid (trans C 18:1-11), as well as oleic (cis C 18:1-9) and elaidic acid (trans C 18:1 9), on basal and glucose-stimulated insulin release. All fatty acids tested increased basal insulin release; however, a significantly lower basal insulin release was demonstrated for cells cultured with 0.3 to 0.4 mmol/L trans vaccenic acid compared to equimolar levels of the cis isomer. GSIS was not changed by cis or trans vaccenic acid or by oleic acid, whereas it was stimulated by 0.3 to 0.4 mmol/L elaidic acid. The mechanisms behind the fatty acid-induced changes in the beta cells have been linked to changes in glucose and fatty acid oxidation. We demonstrated an increased fatty acid oxidation in beta cells after long-term exposure to all of the tested fatty acids. Interestingly, both trans isomers (trans vaccenic and elaidic acid) induced higher fatty acid oxidation than the cis isomers (cis vaccenic and oleic acid, respectively). No changes in glucose oxidation were found when INS-1 cells were cultured with either of the fatty acids. The increased fatty acid oxidation was associated with an increased content of carnitine palmitoyltransferase I (CPT-I) mRNA, but no difference in the content of CPT-I mRNA to the different fatty acids was found. Insulin mRNA expression in beta cells was not affected by the fatty acids. In conclusion, we have demonstrated that the pathological changes in insulin secretion from INS-1 cells to long-term culture with elevated levels of fatty acids are more pronounced for the cis (cis vaccenic acid and oleic acid) rather than the trans isomers (trans vaccenic acid and elaidic acid). We suggest that this, at least in part, may be explained by a lower fatty acid oxidation in cells cultured with the cis compared to the trans fatty acid isomers. Apparently, the difference in fatty acid oxidation was not caused by an increased induction of CPT-I mRNA, nor by changes in glucose oxidation or insulin mRNA in beta cells chronically exposed to the fatty acids. PMID- 15334379 TI - Effect of Pancreas Tonic (an ayurvedic herbal supplement) in type 2 diabetes mellitus. AB - Although there is widespread use of herbal dietary supplements that are believed to benefit type 2 diabetes mellitus, few have been proven to do so in properly designed randomized trials; their efficacy for intermediate-term glucose control remains unclear. Pancreas Tonic is a botanical mixture of traditional Indian Ayurvedic herbs currently available as a dietary supplement. We report the results of a single-center, randomized, double-blind, placebo-controlled 3-month trial of Pancreas Tonic in type 2 diabetic patients inadequately treated with diet/lifestyle or stable doses of sulfonylureas and/or metformin for at least 3 months. Patients with type 2 diabetes for >/= 1 year were entered into 2 strata of hemoglobin A(1c) (HbA(1c)) levels (stratum 1: 8.0% to 9.9%; stratum 2: 10.0% to 12.0%). All subjects began a 1-month single-blind placebo run-in phase, followed by randomization in a 2:1 ratio of active treatment: placebo, to 3 months of double-blind treatment with either Pancreas Tonic or matching placebo (2 capsules 3 times a day). Concurrent oral agents were continued unchanged throughout the study. The primary outcome was the change in HbA(1c) from randomization; results of each stratum were analyzed independently. The baseline characteristics of 36 subjects who completed the study were comparable between treatment groups. Nineteen subjects entered stratum 1 and 17 entered stratum 2. A statistically significant reduction of HbA(1c) from randomization to end-of-study was seen in the stratum 2 subjects (Pancreas Tonic: 10.1% +/- 1.0% to 8.8% +/- 1.9%, P =.004; placebo: 10.8% +/- 1.4% to 11.2% +/- 1.8%, not significant [NS]). No significant HbA(1c) reductions were seen in the stratum 1 subjects. There were no significant treatment-related differences in the fasting plasma glucose (FPG), lipids, body mass index (BMI), body composition, blood pressure, insulin sensitivity estimates using the minimal model, glucose and insulin responses to a meal challenge, quality of life, adverse events, or other safety indices between treatment groups. Pancreas Tonic was well tolerated. Treatment with Pancreas Tonic (2 capsules 3 times per day) for 3 months significantly improved glucose control in type 2 diabetic patients with HbA(1c) levels between 10.0% to 12.0%. This study represents the first properly designed, prospective intervention trial of therapy with an Ayurvedic herbal supplement for intermediate-term glucose control in type 2 diabetes. PMID- 15334380 TI - Production rates of cortisol in men with hypogonadism. AB - Healthy men have a larger endogenous cortisol production rate (PR) than healthy women. To investigate whether this sex-specific difference is maintained in men with low serum testosterone concentrations the endogenous PRs (2 pm to 6 pm) of testosterone, dihydrotestosterone (DHT), and cortisol were simultaneously determined in 10 hypogonadal men. As expected, hypogonadal men were characterized by subnormal PRs of testosterone (19.6 +/- 5.7 microg/h; normal, 180 to 346 microg/h) and of DHT (1.6 +/- 1.1 microg/h; normal, 11 to 20 microg/h). In hypogonadal patients with an intact pituitary-adrenal axis (n = 8), plasma concentrations (7.3 +/- 1.8 microg/dL), metabolic clearance rates (MCRs) (10.0 +/ 4.6 L/h), and endogenous PRs (0.6 +/- 0.2 mg/h) of cortisol were comparable to those seen in eugonadal men. Hence, the sex-specific difference in endogenous cortisol PRs does not depend on the prevailing serum concentrations and on the endogenous PRs of testosterone. PMID- 15334381 TI - Endothelin upregulates the expression of vasopressin V2 mRNA in the inner medullary collecting duct of the rat. AB - Recent studies in our laboratory have demonstrated that bosentan, a mixed endothelin ET(A)/ET(B) receptor antagonist, prevented the upregulation of the arginine vasopressin (AVP) V(2) receptor in the inner medullary collecting duct (IMCD) of cardiomyopathic hamsters. These results suggested that endothelin-1 (ET 1) is involved in the upregulation of AVP V(2) receptors. Studies were performed to detect the effect of ET-1 on the expression of AVP V(2) receptors and the ET receptor mediating these effects within the IMCD of the rat. Rat IMCD tissue was isolated and incubated with the following: ET-1, or ET-1 in combination with ET(A) and ET(B) receptor antagonists BQ-123 and BQ-788, respectively, and sarafotoxin c (S6c), an ET(B) receptor-specific agonist. Tissue samples were then analyzed using quantitative reverse-transcriptase polymerase chain reaction (RT PCR) and Western blotting. ET-1 treatment resulted in increased V(2) mRNA from a control level of 186.8 +/- 15.0 amol/microg total RNA to 430.7 +/- 49.0 amol/microg total RNA (P <.003). ET-1/ET(A) treatment resulted in no significant decrease in V(2) mRNA expression 335.0 +/- 38.0 amol/microg total RNA. Whereas ET 1/ET(B), and ET-1/ET(B)/ET(A) treatment resulted in V(2) mRNA approaching control 256.0 +/- 15.0 amol/microg total RNA, and 215.6 +/- 42.3 amol/microg total RNA. However, ET-3 treatment produced no significant changes in V(2) receptor mRNA expression. Sarafotoxin treatment corroborated both the ET-1 and ET receptor antagonist data, demonstrating striking significant increases in V(2) receptor mRNA and protein expression. S6c treatment increased V(2) mRNA expression from a control level of 199 +/- 17.3 amol/microg total RNA to 284.3 +/- 42.1 amol/microg total RNA (P < 05). Western blotting revealed that changes in V(2) mRNA expression in the various treatment conditions were similar to changes in protein expression. Overall, these data indicate that in the IMCD ET-1 increases AVP V(2) receptor expression and these changes are mediated by the ET(B) receptor. PMID- 15334383 TI - The effects of exercise training on abdominal visceral fat, body composition, and indicators of the metabolic syndrome in postmenopausal women with and without estrogen replacement therapy: the HERITAGE family study. AB - The purpose of this research was to investigate the effects of apriori estrogen replacement therapy (ERT) and endurance exercise training in postmenopausal women on abdominal visceral fat (AFV) and other selected variables related to body composition and the metabolic syndrome (MS). Forty-eight healthy and previously sedentary postmenopausal women (mean age, 54.3 years) who were enrolled in the HERITAGE Family Study (HFS) served as subjects. Of these 48 women, 18 were currently taking ERT and the remaining 30 were taking no supplemental estrogen (NHRT). Computed tomography (CT) scans were used to assess AVF as well as total abdominal fat (TAF) and abdominal subcutaneous fat (ASF). Body mass index (BMI) and waist-to-hip ratios (WHR) were calculated while body fat percentage (%FAT) and total fat mass (FATM) was assessed using underwater weighing. Blood assays for HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), and triglycerides (TG) were conducted at a Centers for Disease Control (CDC) certified laboratory, while blood pressure measurements were assessed using an automated system. All measurements were obtained in duplicate before and after a regimen of endurance exercise training. Analysis of variance (ANOVA) showed AVF to be an average of 31.6 cm(2) less in the women receiving ERT, but lost statistical significance when AVF was adjusted for FATM. Mean values for TAF, ASF, and waist girth were also less in the women receiving ERT, but only waist girth achieved statistical significance. No differences were found in BMI or %FAT, but mean WHR was 5% smaller in the ERT group. Baseline values for HDL-C was higher and LDL-C lower in the ERT group. Prevalence of the MS tended to be greater in the NHRT group, but did not achieve statistical significance. There were no differences in training responses in any of the body composition variables between groups, however, in the ERT group LDL-C decreased with training while TG increased. It was concluded that postmenopausal women taking ERT tended to have lower values of AVF and other indicators of body composition, a more favorable lipid profile, and a slightly reduced risk of the MS when compared with women not taking supplemental hormones. Also exercise training did not improve the overall MS status of either group, as LDL-C status improved in the ERT group while TG decreased in the NHRT group. PMID- 15334382 TI - The beta(2)-adrenergic receptor Arg16-gly polymorphism and interactions involving beta(2)- and beta(3)-adrenergic receptor polymorphisms are associated with variations in longitudinal serum lipid profiles: the Bogalusa Heart Study. AB - We examined the effects of combined genotypes of the beta(2)-adrenergic receptor (AR) Arg(16)-Gly and beta(3)-AR Trp(64)-Arg polymorphisms on longitudinal serum total (T-C) and low-density lipoprotein cholesterol (LDL-C) profiles in 1,198 subjects examined multiple times (6,488 observations) from 1973 to 1996 in the Bogalusa Heart Study, at ages from 4.5 to 38 years. Within 5-year age groups, T-C was significantly (P <.05) higher in beta(2)-AR Arg(16)/Arg(16) homozygotes than in Gly(16) carriers among those 4 to 8 (171.4 +/- 30.0 v 161.5 +/- 27.7 mg/dL), 9 to 13 (167.7 +/- 28.6 v 162.4 +/- 27.4 mg/dL), and 14 to 18 (158.8 +/- 29.6 v 154.7 +/- 27.5 mg/dL) years of age, but not in those 19 to 23, 24 to 28, 29 to 33, or 34 to 38 years of age. The beta(3)-AR polymorphism was not associated with variation in either T-C or LDL-C. In multilevel polynomial growth curve models, the combination of the beta(2)-AR Arg(16)/Arg(16) genotype with either the beta(3)-AR Arg(64)/Arg(64) or Trp(64)/Arg(64) genotypes, denoted AA/AX, was associated with variation in longitudinal T-C (P <.01) and LDL-C (P <.01) profiles. The association between combined beta(2)/beta(3)-AR genotype and lipid profiles differed among race/sex groups, being most marked in black females, in whom the AA/AX combination was associated with higher T-C and LDL-C profiles across all ages. In White males, the AA/AX combination was most strongly associated with higher lipids in adults. In black males and white females, lipid profiles differed little between genotype groups. Our findings suggest that the beta(2)-AR Arg(16)-Gly genotype influences T-C and LDL-C levels in an age specific manner, that it may interact with beta(3)-AR Trp(64)-Arg genotypes to influence longitudinal T-C and LDL-C profiles, and that the effect of combined beta(2)/beta(3)-AR genotypes on T-C and LDL-C profiles may differ among race/sex groups. PMID- 15334384 TI - In vivo regulation of plasma free fatty acids in insulin resistance. AB - Elevated plasma free fatty acid (FFA) concentrations as seen in obesity, insulin resistance, and type 2 diabetes are partly caused by impaired inhibition of intracellular lipolysis in adipose tissue, and this is considered to be part of the insulin resistance syndrome (IRS). Based on predicted insulin resistance at the level of intracellular lipolysis, patients with the IRS would loose weight by disinhibited lipolysis. Since this is not the case in clinical practice, impaired stimulation of intracellular lipolysis must also play a role. We studied acute plasma FFA changes, representing stimulation and inhibition of intracellular adipose tissue lipolysis, in obese patients with IRS and in healthy controls. Thirteen insulin-resistant (IR) subjects (7 men and 6 women) and 10 controls (6 men and 4 women) underwent a mental stress test (20 minutes) preceded by 60 minutes of rest. After mental stress, an oral glucose tolerance test (OGTT) was performed. Baseline FFA levels were higher in IR patients compared to controls (0.59 +/- 0.06 and 0.31 +/- 0.06 mmol/L, respectively; P =.004). During the 20 minutes of mental stress, FFAs increased significantly in IR subjects from 0.55 +/- 0.07 to 0.67 +/- 0.07 mmol/L (P <.001) and from 0.21 +/- 0.04 to 0.36 +/- 0.07 mmol/L in controls (P =.001). Although the absolute change of plasma FFA was not different, the relative increase was lower in IR subjects (28% +/- 7%) compared to controls (89 +/- 24%; P =.02). Despite the more pronounced mean maximal insulin concentration during the OGTT in IR subjects compared to controls (600.0 +/- 126.6 pmol/L and 208.1 +/- 30.0 pmol/L, respectively), the relative decrease of FFAs was lower in IR subjects (11% +/- 5% v 36% +/- 11% in controls after 30 minutes; P =.04). In conclusion, our study shows impaired acute responses of plasma FFAs upon stimulation by mental stress and inhibition by endogenous insulin in insulin resistance in vivo. The presence of both defects helps to understand weight maintenance in insulin resistance. PMID- 15334385 TI - The multiphasic profile of free fatty acids during the intravenous glucose tolerance test is unresponsive to exogenous insulin. AB - As small increments in insulin concentration profoundly affect lipolysis, our goal was to describe the free fatty acid (FFA) profile during the frequently sampled intravenous glucose tolerance test (FSIGT) and determine if both endogenous and exogenous insulin influenced the FFA profile. Thirteen subjects had both a glucose-only (GO-FSIGT) and insulin-modified FSIGT (IM-FSIGT). Both protocols were of 6 hours duration. At baseline an intravenous glucose bolus (0.3 g/kg) was given. In the IM-FSIGT, insulin was infused from 20 to 25 minutes (4 mU/kg. min). Six additional subjects had both an IM-FSIGT and a normal saline study (NS-Study). For the NS-Study, normal saline solution was infused instead of glucose and insulin. Fasting glucose, insulin, FFA and epinephrine concentrations were similar for all tests. Endogenous insulin peaked at 4 +/- 1 minute in both FSIGTs. The mean calculated peak time of exogenous insulin in the IM-FSIGT was 26 +/- 1 minute. Glucose concentrations were lower and epinephrine concentrations higher in the IM-FSIGT versus GO-FSIGT. During the FSIGTs, the FFA time course revealed four distinct phases, which did not differ between protocols. In phase I (0 to 11 minutes), FFA levels remained near basal (491 +/- 183 micromol/L); in phase II (11 to 79 minutes), FFA levels declined achieving a nadir of 139 +/- 63 micromol/L; in phase III (79 to 188 minutes), FFA levels rose linearly and reattained basal levels; and in phase IV (188 to 360 minutes), FFA levels rose above basal and plateaued at 732 +/- 214 micromol/L (P <.001). In the NS-Study, FFA levels remained near baseline (388 +/- 118 mEq/L) until 180 minutes and then trended upward to 618 +/- 258 micromol/L at 360 minutes. FFA concentrations from 180 to 360 minutes did not differ in the IM-FSIGT versus NS-Study. As the 4 FFA phases did not differ between protocols, the insulin effect on FFA levels in the FSIGT can be attributed to endogenous insulin. But the similarity in FFA levels from 180 to 360 minutes in the IM-FSIGT and NS-Study suggests diurnal variation and not a dynamic related to insulin or the FSIGT protocol is responsible for the final suprabasal FFA plateau. PMID- 15334386 TI - Paradoxical changes of muscle glutamine release during hyperinsulinemia euglycemia and hypoglycemia in humans: further evidence for the glucose-glutamine cycle. AB - Insulin suppresses and counterregulatory hormones increase proteolysis. Therefore, if proteolysis were a major factor determining amino acid fluxes in plasma, one would expect release of glutamine into plasma to be suppressed by insulin under euglycemic conditions and to be stimulated under hypoglycemic conditions. However, release of glutamine into plasma remains unaltered or increases during euglycemic hyperinsulinemia and decreases during insulin-induced hypoglycemia. To investigate the mechanisms for these paradoxical observations and the role of skeletal muscle, we infused overnight fasted volunteers with [U (14)C] glutamine and measured release of glutamine into plasma, its removal from plasma, and forearm glutamine net balance, fractional extraction, uptake and release during 4-hour euglycemic ( approximately 5.0 mmol/L, n = 7) and hypoglycemic ( approximately 3.1 mmol/L, n = 8) hyperinsulinemic ( approximately 230 pmol/L) clamp experiments. During the euglycemic clamps, plasma glutamine uptake and release (both P <.05) and forearm muscle glutamine fractional extraction (P <.05), uptake (P <.02) and release (P <.01) all increased, whereas forearm glutamine net balance remained unchanged. The increase in muscle glutamine release (from 1.85 +/- 0.26 to 2.18 +/- 0.30 micromol. kg(-1). min(-1)) accounted for approximately 60% of the increase in total glutamine release into plasma (from 5.54 +/- 0.47 to 6.10 +/- 0.64 micromol. kg(-1). min(-1)) and correlated positively with the increase in muscle glucose uptake (r = 0.80, P <.03). During the hypoglycemic clamps, plasma glutamine uptake and release and forearm glutamine release remained unaltered, but forearm glutamine fractional extraction and uptake decreased approximately 25% (both P <.01) so that forearm glutamine net release increased from 0.37 +/- 0.06 to 0.61 +/- 0.09 micromol. kg( 1). min(-1) (P <.03). We conclude that skeletal muscle is largely responsible for the increased release of glutamine into plasma during euglycemic hyperinsulinemia in humans, and that this may be due to increased conversion of glucose to glutamine as part of the glucose-glutamine cycle; during hypoglycemic hyperinsulinemia decreased glutamine uptake by skeletal muscle may be important for providing substrate for increased glutamine gluconeogenesis. PMID- 15334387 TI - Development of whole-body and skeletal muscle insulin resistance after one day of hindlimb suspension. AB - Hindlimb suspension (HS) of rats is a model of simulated weightlessness and induces dynamic alterations in insulin action. In the present study, the effect of acute (1-day) HS on whole-body glucose tolerance and insulin action on skeletal muscle glucose transport was assessed in juvenile, female Sprague-Dawley rats. Compared to weight-bearing control rats, 1-day HS animals displayed significantly decreased glucose tolerance and diminished whole-body insulin sensitivity. Glucose transport activity in the 1-day unweighted soleus muscle was significantly decreased (P <.05) compared to weight-bearing control muscles both in the absence and presence of insulin (2 mU/mL). Insulin-mediated glucose transport activity in the extensor digitorum longus (EDL) muscles also tended (P =.09) to be lower. There was no change in the protein expression of insulin receptor beta-subunit (IR-beta), insulin receptor substrate-1 (IRS-1), IRS-2, the p85 subunit of phosphatidylinositol-3 kinase (PI3-kinase), Akt, and glucose transporter protein 4 (GLUT-4). The activities of these proteins were also unchanged, as insulin-stimulated IR-beta tyrosine phosphorylation, IRS-1 tyrosine phosphorylation, IRS-1-associated p85, and Akt serine phosphorylation were similar to controls. However, basal Akt phosphorylation was significantly depressed (P <.05) in the 1-day HS soleus. In addition, the protein expression and basal phosphorylation of the stress-activated p38 mitogen-activated protein kinase (p38 MAPK) were significantly elevated (P <.05) in the 1-day unweighted soleus. These results indicate that the development of insulin resistance in the 1-day unweighted soleus is not due to impaired functionality of elements involved in the IR/IRS-1/PI3-kinase/Akt signaling pathway. However, activation of p38 MAPK may play a role in this response. PMID- 15334388 TI - Adiponectin gene polymorphism (G276T) is not associated with incipient diabetic nephropathy in Japanese type 2 diabetic patients. AB - A single-nucleotide polymorphism (SNP) G276T in the adiponectin gene has been associated with lower plasma adiponectin levels and insulin resistance, which are related to the prevalence of type 2 diabetes or diabetic complications of macroangiopathy. We performed a case-control study to examine whether the SNP276 of the adiponectin gene was also related to early diabetic nephropathy. SNP276 was examined with genomic DNA obtained from 108 type 2 diabetic patients with microalbuminuria (urinary albumin creatinine ratio [ACR] between 30 mg/g x Cr and 300 mg/g x Cr; case subjects), and 208 patients with normoalbuminuria (ACR < 30 mg/g x Cr; control subjects). The genotype distribution and G allele frequency of SNP276 in the case subjects (0.71) did not significantly differ from the control subjects (0.69). There were no differences among the genotypes of the adiponectin gene regarding age, duration of diabetes, body mass index (BMI), hemoglobin A(1c) (HbA(1c)), serum lipids, serum creatinine, and plasma adiponectin levels. These data suggest that SNP276 of the adiponectin gene is not an independent risk factor for incipient diabetic nephropathy in Japanese type 2 diabetic patients. PMID- 15334389 TI - Effect of pramlintide on symptom, catecholamine, and glucagon responses to hypoglycemia in healthy subjects. AB - Pramlintide is an analog of the human glucoregulatory hormone amylin. Previous studies have shown no clear evidence that pramlintide modifies the response to insulin-induced hypoglycemia; however, a detailed assessment of responses at hypoglycemic thresholds has not been conducted. To further test the effect of pramlintide on symptom, catecholamine, and glucagon responses, a 3-step hypoglycemic clamp was investigated in healthy volunteers. In a randomized, double-blind, placebo-controlled, crossover study, 18 healthy subjects without diabetes received subcutaneous premeal injections of either placebo or 60 microg pramlintide 3 times daily for 5 consecutive days. On day 6, subjects received study drug with breakfast and, after a 7-hour fast, were connected to a Biostator for a 3-step, 3-hour clamp experiment (insulin infusion rate: 1.0 mU/kg/min; blood glucose targets: 70, 55, and 45 mg/dL). An intravenous (IV) infusion of pramlintide (16 microg/h) or placebo was initiated at t = 60 minutes. At the end of each 60-minute clamp step, autonomic (sweating, palpitations, hunger, etc) and neuroglycopenic (confusion, headache, odd behavior, etc) symptoms were assessed using a validated visual analog scale questionnaire. Blood samples were collected at 30-minute intervals for measurement of plasma glucose, insulin, pramlintide, catecholamine, and glucagon concentrations. Intraindividual and group mean responses showed that autonomic symptoms and plasma catecholamine and glucagon concentrations increased progressively during the clamp, with no discernible differences between pramlintide and placebo treatments. Group means for catecholamines at 60 minutes were: epinephrine 233 +/- 42, 892 +/- 85, 2,340 +/- 302 and 202 +/- 25, 774 +/- 114, 2,751 +/- 404 pg/mL and norepinephrine 1,138 +/- 86, 1,236 +/- 77, 1,721 +/- 158 and 1,278 +/- 108, 1,259 +/- 109, 1,580 +/-136 pg/mL (+/- SEM) for placebo- and pramlintide-treated groups at 70, 55, and 45 mg/dL glucose, respectively. Group means for glucagon were 72 +/- 6.3, 98 +/- 11.1, 130 +/- 14.7 and 63 +/- 3.6, 92 +/- 9.4, 120 +/- 16.0 pmol/L (+/- SEM) for placebo- and pramlintide-treated groups at 70, 55, and 45 mg/dL glucose, respectively. These results showed that pramlintide did not impair the symptom, catecholamine, and glucagon responses to insulin-induced hypoglycemia in healthy subjects. PMID- 15334390 TI - Exercise training increases glycogen synthase activity and GLUT4 expression but not insulin signaling in overweight nondiabetic and type 2 diabetic subjects. AB - Exercise training improves insulin sensitivity in subjects with and without type 2 diabetes. However, the mechanism by which this occurs is unclear. The present study was undertaken to determine how improved insulin signaling, GLUT4 expression, and glycogen synthase activity contribute to this improvement. Euglycemic clamps with indirect calorimetry and muscle biopsies were performed before and after 8 weeks of exercise training in 16 insulin-resistant nondiabetic subjects and 6 type 2 diabetic patients. Training increased peak aerobic capacity (Vo(2peak)) in both nondiabetic (from 34 +/- 2 to 39 +/- 2 mL O(2)/kg fat-free mass [FFM]/min, 14% +/- 2%, P <.001) and diabetic (from 26 +/- 3 to 34 +/- 3 mL O(2)/kg FFM/min, 32% +/- 4%) subjects. Training also increased insulin-stimulated glucose disposal in nondiabetic (from 6.2 +/- 0.5 to 7.1 +/- 0.7 mg/kg FFM/min) and diabetic subjects (from 4.3 +/- 0.6 to 5.5 +/- 0.6 mg/kg FFM/min). Total glycogen synthase activity was increased by 46% +/- 17% and 45% +/- 12% in nondiabetic and diabetic subjects, respectively, in response to training (P <.01 v before training). Moreover, after training, glycogen synthase fractional velocity was correlated with insulin-stimulated glucose storage (r = 0.53, P <.05) and the training-induced improvement in glucose disposal was accounted for primarily by increased insulin-stimulated glucose storage. Training also increased GLUT4 protein by 38% +/- 8% and 22% +/- 10% in nondiabetic and diabetic subjects, respectively (P <.05 v. before training). Akt protein expression, which was decreased by 29% +/- 3% (P <.05) in the diabetic subjects before training (compared to the nondiabetics), increased significantly in both groups (P <.001). In contrast, exercise training did not enhance the ability of insulin to stimulate insulin receptor substrate-1 (IRS-1)-associated phosphatidylinositol 3 (PI 3)-kinase activity. The present data are consistent with a working model whereby 8 weeks of exercise training increases insulin-stimulated glucose disposal primarily by increasing GLUT4 protein expression without enhancing insulin-stimulated PI 3-kinase signaling, and that once the glucose enters the myocyte, increased glycogen synthase activity preferentially shunts it into glycogen synthesis. PMID- 15334391 TI - Preclinical changes in the mechanical properties of abdominal aorta in obese children. AB - Obesity in childhood has been associated with the development of early cardiovascular abnormalities. The aim of the present study was to investigate whether preclinical functional changes are detectable in the abdominal aorta of obese children. One hundred consecutively seen obese children and 50 healthy controls were studied. The groups were matched in terms of age and gender. The pulsatile wall-motion of the abdominal aorta was determined using a B-mode ultrasound technique. The following mechanical property parameters were measured or computed: lumen diastolic and systolic diameters, relative aortic strain, elastic modulus, and stiffness. Compared to controls, obese children had higher blood pressure values and higher concentrations of total cholesterol, triglycerides, insulin, and C-reactive protein. Homeostasis model assessment (HOMA) score, a parameter of insulin resistance, was significantly higher in obese children than in controls (3.2 +/- 1.9 v 1.4 +/- 0.5, P <.001). Aortic mechanical parameters were significantly different in obese children as compared to controls: stiffness was higher (3.00 +/- 1.45 v 2.22 +/- 0.87, P <.001) as was elastic modulus (0.38 +/- 0.18 v 0.24 +/- 0.10 N/m(2), P <.001). Obese girls with insulin resistance (ie, in the highest tertile of HOMA, >3.7) had increased aortic stiffness (3.79 +/- 2.25) compared to obese girls in the lowest tertiles of HOMA (2.67 +/- 1.09, P =.045), even after adjustment for traditional cardiovascular risk factors (P =.031). The present findings suggest that preclinical changes in the aortic elastic properties are detectable in obese children. Insulin resistance seems to play an important role in the increased rigidity of the aortic wall in obese girls. PMID- 15334392 TI - Mandible fracture patterns: a suburban trauma center experience. AB - PURPOSE: Mandible fractures are among the most frequently seen injuries in the trauma center setting. Recent shifts in the mechanism and age distribution of patients sustaining these injuries are well documented. This study attempts to define current, predictable patterns of fracture based on patient characteristics and mechanism of injury. MATERIAL AND METHODS: The charts of 134 patients with 225 mandible fractures treated over a 7-year period by the Otolaryngology-Head and Neck Surgery, Plastic and Reconstructive Surgery and Oral-Maxillofacial Surgery services, our institution, were retrospectively reviewed. Patients were categorized based on age, mechanism of fracture, and anatomic location of fracture. Multivariate analysis of data was performed to determine significant relationships among groups. RESULTS: Violent crimes such as assault and gunshot wounds accounted for the majority of fractures (50%) in this study, with motor vehicle accidents less likely (29%). Overall, parasymphyseal fractures were most frequent (35%), whereas angle and body fractures were also common (15% and 21%, respectively). There was a statistically significant association of motor vehicle accidents with parasymphyseal fractures (45%), and gunshot wounds with body fractures (36%), whereas assault victims had a higher than predicted frequency of angle fractures (27%) and fewer parasymphyseal fractures (19%). Patients aged 17 to 30 were more likely to suffer from gunshot wounds, whereas older adults (age 31-50) were more likely to be assault victims. Patients over age 50 suffered fractures from falls at a higher than expected rate. Although children and young adults seemed to suffer more parasymphyseal fractures and older adults body fractures, these correlations failed to show statistical significance. Parasymphyseal fractures were most frequently associated with fractures at other sites within the mandible, ipsilateral body fractures being the most common. CONCLUSIONS: Updated data on the association of patient age and mechanism of injury with fracture pattern can guide treating physicians in anticipating and diagnosing traumatic mandible fractures. PMID- 15334393 TI - Synchronous thyroid pathology in patients presenting with primary hyperparathyroidism. AB - BACKGROUND: Thyroid and parathyroid pathology may coexist; classically, this is described in multiple endocrine neoplasia. Other nonsyndromic scenarios result in pathologies of these endocrine organs coexisting. This can create a challenge in the clinical decision making and management of these patients. It is optimal to deal with both problems in one operative procedure, if surgery is required. METHODS: A retrospective review of cases of synchronous thyroid and parathyroid pathology that were managed surgically over a 3-year period by a single endocrine surgeon at a single university medical center. RESULTS: Seventeen cases of synchronous thyroid and parathyroid disease were discovered. All required surgical management of thyroid and parathyroid conditions. Reasons included removal of thyroid goiter for access to an adenoma, worrisome thyroid lesions coexistent with a parathyroid adenoma, and an intrathyroid parathyroid adenoma. Cases of incidental discovery of a second endocrine condition while the first was evaluated will be reported. CONCLUSIONS: A head and neck endocrine surgeon needs to be aware of the possible coexistence of thyroid and parathyroid pathologies so that, when encountered, they can be safely and efficiently managed in a single procedure. PMID- 15334394 TI - Effects of gentamicin on sensorineural elements of the cochlea in human temporal bones. AB - PURPOSE: Gentamicin is commonly prescribed for treatment of gram-negative bacterial infections. It has, however, been known to have vestibulotoxic and ototoxic effects in humans. Previous studies were confounded by the use of other types of ototoxic drugs, higher than therapeutic doses, and prolonged time periods between gentamicin administration and death. The purpose of this study is to observe the effects of parenteral gentamicin administration on the sensorineural elements of the cochlea in which the previously described factors did not exist. MATERIALS AND METHODS: Temporal bone specimens were divided into 2 groups. Group 1 included 16 "normal" temporal bones with no history or histopathologic findings of otologic disease or ototoxic drug administration, and group 2 included 17 temporal bones that received gentamicin treatment within 6 months before death. Temporal bones were excluded from group 2 if patients had histopathologic findings or history of otologic disease and/or other ototoxic drugs. All temporal bones were examined under light microscopy. RESULTS: The percentages of intact outer hair cells in the basal turn was significantly greater in group 1 compared with group 2. There was no significant difference in hair cell number between the groups in the other turns. There was no significant difference in the number of spiral ganglion cells between groups in any turn. CONCLUSION: Although gentamicin is primarily considered to be vestibulotoxic, this study showed that gentamicin can damage sensorineural elements of the cochlea, even in younger patients. During the administration of this drug, hearing of the patient should be carefully monitored. PMID- 15334396 TI - The microbiology and antimicrobial resistance patterns in chronic rhinosinusitis. AB - OBJECTIVES: The purpose of this study was to review the microbiology of chronic rhinosinusitis (CRS) in patients undergoing endoscopic sinus surgery (ESS) and comment on antimicrobial resistance trends. METHODS: A retrospective review of 101 patients undergoing ESS during the period of 1997 to 2001 was performed. Patients were divided into groups based on their surgical history. Fifty-five patients without prior ESS history were placed in the primary group; 46 patients who had undergone prior ESS were placed in the revision group. Intraoperative microbiology culture data were reviewed and antimicrobial resistance data analyzed. RESULTS: Data on 101 patients were analyzed. There were 182 total cultures sent, yielding 257 isolates. The most common isolates were coagulase negative Staphylococcus (SCN) (45% of cultures), gram-negative rods (25% of cultures), and Staphylococcus aureus (24% of cultures). Pseudomonas aeruginosa was isolated in 9% of cultures. When comparing the 2 patient groups, we did not find consistent trends in the differences in the prevalence of these isolates. Antimicrobial resistance for SCN (P = .01) and S aureus (P < .001) was greater in the revision surgery. Overall, 62% of patients were found to have at least 1 isolate with decreased antibiotic sensitivity. CONCLUSION: The most prevalent microorganisms in patients with CRS are SCN, S aureus, and gram-negative rods. Perhaps more importantly, the antimicrobial sensitivities of these microorganisms appear to be a growing problem. These findings suggest increased antimicrobial resistance in patients undergoing revision ESS when compared with patients undergoing surgery for the first time. PMID- 15334395 TI - Glutathione S-transferase M1, T1, and P1 gene polymorphism in laryngeal squamous cell carcinoma. AB - PURPOSE: To investigate the influence of the glutathione S-transferase (GST) M1, T1, and P1 genotypes on the laryngeal squamous cell carcinoma risk. MATERIAL AND METHODS: The study group consisted of 42 white patients with laryngeal squamous cell carcinoma (39 of them were male, mean age: 53, range: 37-67 and 3 of them were female, mean age: 47, range: 32-55) and 89 control subjects (nonsmokers = 47, smokers = 42) (58 male and 31 female, mean age: 51, range: 30-72). DNA samples were isolated from blood samples using high pure polymerase chain reaction (PCR) Template Preparation Kit. The detection of GST T1, GST M1, and P1 polymorphisms were detected by using real-time PCR. RESULTS: Gene polymorphisms at GST M1 and P1 were not significantly different in patient and control groups. However, GST T1 null type significantly increased in laryngeal cancer patients when compared with the nonsmoking controls (P =.04). CONCLUSIONS: There was a significant association between GST T1 null genotype and laryngeal squamous cell carcinoma. However the potential role of GSTs as markers of susceptibility to laryngeal carcinoma needs further studies in a larger number of patients. PMID- 15334397 TI - Management of otogenic lateral sinus thrombosis. AB - PURPOSE: Lateral sinus thrombosis (LST) is a rare but potentially devastating complication of otitis media. We review the clinical presentation, evaluation, management, operative findings, and outcomes of this serious complication. MATERIAL AND METHODS: A retrospective chart review was performed at a teaching hospital of all patients diagnosed with an otogenic lateral sinus thrombosis between 1992 and 2002. Data on patients with otogenic LST were collected and analyzed. RESULTS: Eleven patients were identified with a diagnosis of lateral sinus thrombosis, and all had a chronic otitis media. The most common presenting symptoms were headache, otalgia, and fever. Radiologic evaluation included computed tomography scan, magnetic resonance imaging, or magnetic resonance venography. All patients had radiographic evidence of LST preoperatively. The thrombosed sinus was on the right side in 6 patients and on the left side in 5 patients. The majority of patients (8 of 11) had a second concomitant intracranial complication. All patients underwent a mastoidectomy. The thrombus was removal in 7 cases, whereas only needle aspiration of the sinus was performed in 4 cases. Gram-negative bacilli and anaerobes were the most common organisms. There were no deaths but sequelae included VI nerve palsy, ataxia, and dead ear. CONCLUSION: LST as a complication of otogenic infections may still pose a serious threat that warrants immediate attention and care. It is frequently associated with other intracranial complications. Contrast-enhanced computed tomography scan and magnetic resonance imaging plays a major role in determining diagnosis and treatment plans. The availability of broad-spectrum antibiotics has improved our management significantly. Conservative surgical intervention including eradication of all perisinus infection and needle aspiration of the sinus seems to be effective. PMID- 15334398 TI - A preoperative sleep study with nasal airway occlusion in pharyngeal flap surgery. AB - PURPOSE: Obstructive sleep apnea is a major complication of pharyngeal flap surgery. The purpose of the present study is to predict preoperatively the risk of upper airway obstruction after surgery. MATERIAL AND METHODS: We performed an overnight sleep study preoperatively and postoperatively in 16 pediatric patients considered for pharyngeal flap surgery. Preoperative sleep study was done for two nights, once in normal breathing condition and once with complete nasal occlusion by packing of nostril with tampon gauze. RESULTS: In preoperative sleep recordings in normal breathing condition, all subjects had a normal apnea hypopnea index (AHI) less than 5/h. In preoperative recording with complete nasal occlusion, five patients exceeded 5/h in AHI. In particular, for two patients who had AHI higher than 15/h, we gave up a surgery in one case and performed pharyngeal flap operation for the other following a tracheotomy for severe disturbance of oral breathing. The remaining 14 subjects underwent surgery without airway obstructive complications. There was strong correlation between preoperative AHI with nasal tampon gauze and AHI at two weeks postoperatively (r = 0.88 P < .0001). There was no significant correlation between preoperative AHI in normal breathing condition and postoperative AHI (P > .05). CONCLUSIONS: These results exhibit preoperative sleep study with complete nasal airway occlusion represent postoperative breathing condition well during early postoperative period. Preoperative sleep study with complete nasal airway occlusion with nasal tampons could be useful for predicting the risk of upper airway obstruction secondary to pharyngeal flap surgery. PMID- 15334399 TI - Congenital absence or hypoplasia of the carotid artery: radioclinical issues. AB - PURPOSE: Congenital anomaly of the carotid artery is a rare abnormality. It is usually discovered incidentally by color Doppler carotid sonography, angiography, computed tomography (CT), or magnetic resonance imaging (MRI) of the head and neck taken for some other reason. Most patients are not symptomatic because of sufficient cerebral circulation supplied to the defective area by the communicating arteries of the circle of Willis, intercavernous anastomosis, communicating arteries from external carotid artery, and by persistent embryologic arteries to the carotid artery territory. However, sometimes, this anatomic variation may eventually lead to some clinical signs and symptoms in particular circumstances in the head and neck of which surgeons are unaware. A retrospective study was designed to emphasize the characteristic radiologic and clinical picture in patients with the congenital absence (agenesis and aplasia) or hypoplasia of the carotid artery, to delineate the associated abnormalities and existing collateral vessels, and to find out its incidence. MATERIAL AND METHODS: Five thousand one hundred cerebral MRI and/or catheter angiograms performed between February 1988 and March 2002 were reviewed for carotid artery abnormality. RESULTS: Seven patients with congenital absence or hypoplasia (4 of these patients were presented with hypoplasia of internal carotid artery [ICA], 3 with absence of ICA) of ICA were identified (0.13%). The radiologic and clinical study of 5 patients with unilateral (3 of these patients were presented with hypoplasia and 2 with absence of ICA) and 2 patients with bilateral (1 was presented with absence of ICA, whereas the other was hypoplasia) congenital abnormality with absence or hypoplasia of ICA demonstrate that those patients are usually asymptomatic and they are diagnosed incidentally. CONCLUSION: It has been concluded that the combined use of magnetic resonance angiography and CT scanning of the skull base may disclose small but patent ICA. Collateral vessels seem to be usual in such cases, but they may be prominent in cases of acquired vascular occlusion, or increased hemodynamic pressure in dysplastic changes in collateral arteries are known causes of aneurysms. The main vascular supply for the brain in patients with congenitally small (hypoplasia) or absent (agenesis or aplasia) ICA is the vertebrobasilar system in bilateral cases. However, contralateral carotid vessel is the dominant arterial supply for unilateral cases, which has to be borne in mind in surgical interventions to the involved side. PMID- 15334400 TI - Primary sphenoid sinus esthesioneuroblastoma. AB - Esthesioneuroblastoma is a rare malignant tumor of undifferentiated neuroectodermal origin. Because these tumors are derived from olfactory epithelium, most cases arise in the superior nasal cavity at the level of the cribriform niche. The development of an esthesioneuroblastoma outside of the region in which olfactory epithelium exists is exceedingly rare. We report a rare case of a primary sphenoid sinus esthesioneuroblastoma and discuss the pathogenesis, clinical presentation, and treatment of this rare entity. PMID- 15334401 TI - Two consecutive cases of cerebrospinal fluid rhinorrhea after septoplasty operation. AB - Our aim in this work is to define the importance of anatomical knowledge in septoplasty operation and to prevent complications. Septoplasty is one of the most common operations in otorhinolaryngology to treat the nasal obstruction caused by septal deviation. During and after septoplasty, there are some recorded complications, such as hemorrhage, hematoma, septal abscess, septal perforation, saddle nose, infection, anosmia, visual disturbances, cavernous sinus thrombosis, meningitis, pneumoencephalos, subarachnoid hemorrhage, subdural empyma, brain abscess, periorbital emphysema, toxic shock syndrome, and cerebrospinal fluid (CSF) rhinorrhea. Some of the complications are rare, but their results are life threatening. We report a rare complication of septoplasty CSF rhinorrhea. Two consecutive cases of CSF fistulas after septoplasty operation are presented. Both of the cases were treated endoscopically. The possible mechanisms and different treatment options are discussed. Prevention of the CSF fistula in septoplasty is more important than its treatment. Realizing the anatomic variations and gentle manipulation at the ethmoid roof is essential. PMID- 15334402 TI - Lateral sinus thrombosis associated with zoster sine herpete. AB - Herpes zoster results from reactivation of the varicella zoster virus (VZV). Zoster sine herpete (ZSH) is an uncommon manifestation of VZV infection and presents with similar symptoms but without the vesicular rash. We describe an unusual case of lateral sinus thrombosis (LST) that developed during the clinical course of ZSH in the C2 distribution. A 55-year-old woman presented with a 3-day history of left temporal and postauricular pain, nausea, vomiting, and mild photophobia. She denied otalgia, otorrhea, and hearing loss. Examination revealed hyperesthesia in the left C2 nerve root distribution without evidence of herpetic rash. A computed tomography scan showed minimal fluid in the left mastoid cavity (not mastoiditis) and thrombus within the left lateral and sigmoid dural sinus. Magnetic resonance imaging and magnetic resonance angiogram confirmed these findings. Laboratory studies revealed elevated neurotrophic immunoglobulin G levels to VZV. Hypercoagulable studies were normal. She was subsequently treated with Neurontin, acyclovir, and anticoagulation. Her symptoms improved, and she was discharged 3 days later. LST is generally a complication of middle ear infection. Nonseptic LST, however, may result from dehydration, oral contraceptive use, coagulopathy, or thyroid disease. This unusual case raises the suspicion that thrombosis resulted from VZV associated thrombophlebitis in the ipsilateral cerebral venous sinuses along the second cervical nerve root distribution. A high index of suspicion is necessary in such cases so that a different treatment course can be identified and antiviral medication initiated promptly. PMID- 15334403 TI - Bilateral vocal cord paralysis secondary to esophageal compression. AB - Bilateral vocal cord paralysis is most commonly caused by trauma, malignancy, and neurologic disorders. Cases secondary to esophageal compression of the recurrent laryngeal nerves are rare. We report a patient admitted with an exacerbation of achalasia who developed acute respiratory distress from bilateral immobile vocal cords. Imaging studies revealed impressive dilation of the cervical esophagus causing compression of both recurrent laryngeal nerves. After securing the airway and decompression of the esophagus, mobility of the vocal cords returned within 1 week. This case shows the importance of a careful airway workup in patients with esophageal distention. Early decompression may prevent permanent recurrent laryngeal nerve injury and airway obstruction. PMID- 15334404 TI - Report of an obstructive goiter and its surgical treatment during delivery. AB - We report a case of a morbidly obese young woman in her third trimester of pregnancy presenting with a history of goiter and respiratory disease. The recent history of this patient was significant for worsening respiratory symptoms over a period of 2 weeks, and, on presentation at 36 weeks gestation, she was stridorous, dyspneic at rest, and had a hoarse voice. Evaluation revealed a morbidly obese individual with a large goiter. She was biochemically euthyroid. Fiberoptic laryngoscopy revealed a left true vocal cord paresis, and ultrasound evaluation was significant for diffuse multinodular enlargement, with each lobe measuring greater than 10 cm and the isthmus measuring 5. Pulmonary function testing revealed a significant degree of upper airway obstruction without significant lower airway disease. Given the patient's clinical signs and symptoms, her tenuous airway, poor candidacy for urgent tracheotomy, and her proximity to delivery, it was agreed that the patient should undergo elective cesarean section and at its completion undergo subtotal thyroidectomy for the obstructive goiter. PMID- 15334405 TI - High-located aberrant innominate artery: an unusual cause of serious hemorrhage of percutaneous tracheotomy. AB - An aberrant innominate artery located high was observed during a cadaveric neck dissection. To our knowledge, an innominate artery crossing the 4th and 5th tracheal rings has rarely been reported in available literature. Knowledge of this anomaly is important for those who are involved with percutaneous procedures in the neck to avoid major complications. PMID- 15334406 TI - Aortoarteritis and sensorineural hearing loss in an adolescent black male. AB - We describe an adolescent male having sudden onset of sensorineural hearing loss. Evaluation led to the discovery of marked aortoarteritis. This patient exhibited findings of both Takayasu arteritis and Cogan syndrome. This report emphasizes the importance of ensuring a thorough systemic evaluation in individuals having sudden hearing loss. PMID- 15334407 TI - Ossifying fibroma of the middle turbinate: a case report. AB - Ossifying fibromas (OFs) are rare, benign, nonaggressive fibroosseous tumors that are commonly seen in head and neck region. They show aggressive pattern when the midface and paranasal sinuses are involved. We report a 28-year-old woman with OF of the middle turbinate. Computed tomography images of the nasal cavity showed monostotic hyperdense lesion confined to middle turbinate. En bloc excision was performed. Histological examination confirmed the diagnosis as OF. Our case is of particular interest because it is, to our knowledge, the first documented case of OF arising from the middle turbinate. PMID- 15334408 TI - Thermodynamic analysis of product formation in mesophilic acidogenesis of lactose. AB - Thermodynamic analysis on the acidogenesis of lactose was performed to evaluate the different acidogenic patterns and mechanisms by using Gibbs free energy calculation. Batch acidogenesis of lactose was investigated by using an enriched culture at 37 degrees C, pH 5.5 and varied substrate levels. In addition to usual acidogenic products, i-butyrate, valerate, i-valerate, caproate, and propanol were also produced at a significant level. Thermodynamic analysis shows that valerate might be formed through the reaction requiring hydrogen as electron donor and consuming of propionate and carbon dioxide. Caproate was most likely produced directly from butyrate, hydrogen, and carbon dioxide. The minimum amount of Gibbs free energies needed to sustain isomerization of butyrate and valerate were approximately 5.7-5.8 and 4.5-4.6 kJ/mol, respectively. Propanol was produced from acetate, hydrogen, and carbon dioxide with a minimum amount of Gibbs free energy of 41.8-42.0 kJ/mol. Formation of butanol was controlled more by substrate level or population dynamics than by thermodynamics. PMID- 15334409 TI - Microbial population dynamics during start-up and overload conditions of anaerobic digesters treating municipal solid waste and sewage sludge. AB - Microbial population dynamics were investigated during start-up and during periods of overload conditions in anaerobic co-digesters treating municipal solid waste and sewage sludge. Changes in community structure were monitored using ribosomal RNA-based oligonucleotide probe hybridization to measure the abundance of syntrophic propionate-oxidizing bacteria (SPOB), saturated fatty acid-beta oxidizing syntrophs (SFAS), and methanogens. These changes were linked to traditional performance parameters such as biogas production and volatile fatty acid (VFA) concentrations. Digesters with high levels of Archaea started up successfully. Methanosaeta concilii was the dominant aceticlastic methanogen in these systems. In contrast, digesters that experienced a difficult start-up period had lower levels of Archaea with proportionally more abundant Methanosarcina spp. Syntrophic propionate-oxidizing bacteria and saturated fatty acid-beta-oxidizing syntrophs were present at low levels in all digesters, and SPOB appeared to play a role in stabilizing propionate levels during start-up of one digester. Digesters with a history of poor performance tolerated a severe organic overload event better than digesters that had previously performed well. It is hypothesized that higher levels of SPOB and SFAS and their methanogenic partners in previously unstable digesters are responsible for this behavior. PMID- 15334410 TI - Real-time detection of 13C NMR labeling kinetics in perfused EMT6 mouse mammary tumor cells and betaHC9 mouse insulinomas. AB - A method was developed for obtaining high signal-to-noise 13C NMR spectra of intracellular compounds in metabolically active cultured cells. The method allows TCA cycle labeling kinetics to be determined in real time without significant oxygen transport limitations. Cells were immobilized on the surface of nonporous microcarriers that were either uncoated or coated with polypeptides and used in a 12-cm3 packed bed. The methods were tested with two EMT6 mouse mammary tumor cell lines, one strongly adherent and the other moderately adherent, and a weakly adherent mouse insulinoma line (betaHC9). For both EMT6 lines, NTP and oxygen consumption measurements indicated that the number of cells in the spectrometer ranged from 6 x 10(8) to 1 x 10(9). During infusion of [1-13C]glucose, labeling in C-4 glutamate (indicative of flux into the first half of the TCA cycle) could be detected with 15-min resolution. However, labeling for C-3 and C-2 glutamate (indicative of complete TCA cycle activity) was fivefold lower and difficult to quantify. To increase TCA cycle labeling, cells were infused with medium containing [1,6-13C2]glucose. A 2.5-fold increase was observed in C-4 glutamate labeling and C-3 and C-2 glutamate labeling could be monitored with 30-min resolution. Citrate synthase activity was indirectly detected in real time, as [3,4-13C2]glutamate was formed from [2-13C]oxaloacetate and [2-13C]acetate (of acetyl-CoA). Cell mass levels observed with betaHC9 cells were somewhat lower. However, the 13C S/N was sufficient to allow real-time monitoring of the response of intracellular metabolite labeling to a step change in glucose and a combined glutamine/serum pulse. PMID- 15334411 TI - Enhancement of tyrosinase inhibition of the extract of Veratrum patulum using cellulase. AB - Inhibitors of melanin biosynthesis were screened by using three different methods. The extract of Veratrum patulum contains hydroxystilbene compounds that are potent tyrosinase inhibitors. We evaluated the enzyme inhibitory property on the mushroom tyrosinase of hydroxystilbene compounds including resveratrol, oxyresveratrol, and their analogs. Biotransformation using cellulase of the whole extract brought about an increase in the inhibitory activity of the products on mushroom tyrosinase. The enhancement of tyrosinase inhibition is supposed to increase the concentration of aglycon, which has superior inhibitory activity to its glycoside. Eventually, melanin biosynthesis was inhibited by the enhanced tyrosinase inhibitory activity of the extract. This result indicated that deglycosylation of stilbene compounds has exerted more effective inhibition on the enzyme than that of the unprocessed plant extract. PMID- 15334412 TI - An impedance study on admiralty brass dezincification originated by microbiologically influenced corrosion. AB - In this article we describe a field study of biofouling and microbiologically influenced corrosion (MIC) of admiralty brass heat exchanger tubes in contact with running fresh water on the river Tagus close to Almaraz nuclear power plant in Spain. Dezincification originated by biofouling and MIC was studied using impedance, polarization resistance, gravimetric, scanning electron microscopy (SEM), and X-ray diffraction (XRD) measurements. Close correlation was observed between the biofilms formed and the corrosion process (dezincification) using the different experimental techniques. Impedance data showed a capacitive behavior including two time constants. Kramers-Kronig (KK) transforms were used to validate impedance data. The admiralty tubes' impedance data satisfied the KK relations. PMID- 15334413 TI - Hybrid pancreatic tissue substitute consisting of recombinant insulin-secreting cells and glucose-responsive material. AB - Insulin-dependent diabetes is a serious pathological condition, currently treated by blood glucose monitoring and daily insulin injections, which, however, do not prevent long-term complications. A tissue-engineered pancreatic substitute has the potential to provide a more physiologic, less invasive, and potentially less costly treatment of the disease. A major issue in developing such a substitute is the cells being used. Nonpancreatic cells, retrieved from the same patient and genetically engineered to secrete insulin constitutively or with some glucose responsiveness, offer the significant advantages of being immune-acceptable and relaxing the tissue availability limitations, which exist with islets from cadaveric donors. These cells, however, do not have insulin secretion dynamics appropriate for restoration of euglycemia in higher animals and, eventually, humans. In this study, we present the concept of a hybrid pancreatic substitute consisting of such cells sequestered in a material exhibiting glucose-dependent changes of its permeability to insulin. A Concanavalin A-glycogen material sandwiched between two polycarbonate membranes and exhibiting glucose-dependent sol-gel transformations was used. Rates of insulin transport through this material in gel and sol forms were characterized for both FITC-labeled insulin in solution and insulin secreted by betaTC3 mouse insulinoma cells. Effective diffusivities through sol were found to be up to 3.5-fold higher than through the gel state of the material. A mathematical model of a hybrid construct was formulated and analyzed to simulate the secretory behavior in response to step ups and downs in the surrounding glucose concentration. The experimental and modeling studies indicate that a hybrid pancreatic substitute consisting of constitutively secreting cells and glucose-responsive material has the potential to provide a more physiologic regulation of insulin release than the cells by themselves or in an inert material. PMID- 15334414 TI - Modeling hexavalent chromium removal in a Bacillus sp. fixed-film bioreactor. AB - A one-dimensional diffusion-reaction model was developed to simulate Cr(VI) reduction in a Bacillus sp. pure culture biofilm reactor with glucose as a sole supplied carbon and energy source. Substrate utilization and Cr(VI) reduction in the biofilm was best represented by a system of (second-order) partial differential equations (PDEs). The PDE system was solved by the (fourth-order) Runge-Kutta method adjusted for mass transport resistance using the (second order) Crank-Nicholson and Backward Euler finite difference methods. A heuristic procedure (genetic search algorithm) was used to find global optimum values of Cr(VI) reduction and substrate utilization rate kinetic parameters. The fixed film bioreactor system yielded higher values of the maximum specific Cr(VI) reduction rate coefficient and Cr(VI) reduction capacity (kmc = 0.062 1/h, and Rc = 0.13 mg/mg, respectively) than previously determined in batch reactors (kmc = 0.022 1/h and Rc = 0.012 mg/mg). The model predicted effluent Cr(VI) concentration well with 98.9% confidence (sigmay2 = 2.37 mg2/L2, N = 119) and effluent glucose with 96.4 % confidence (sigmay(w)2 = 5402 mg2/L2, N = 121, w = 100) over a wide range of Cr(VI) loadings (10-498 mg Cr(VI)/L/d). PMID- 15334415 TI - Real time quantitative PCR as a method to evaluate xenotropic murine leukemia virus removal during pharmaceutical protein purification. AB - Chinese hamster ovary cells used for pharmaceutical protein production express noninfectious retrovirus-like particles. To assure the safety of pharmaceutical proteins, validation of the ability of manufacturing processes to clear retrovirus-like particles is required for product registration. Xenotropic murine leukemia virus (X-MuLV) is often used as a model virus for clearance studies. Traditionally, cell-based infectivity assay has been the standard virus quantification method. In this article, a real time quantitative PCR (Q-PCR) method has been developed for X-MuLV detection/quantification. This method provides accurate and reproducible quantification of X-MuLV particle RNA (pRNA) over a linear dynamic range of at least 100,000-fold with a quantification limit of approximately 1.5 pRNA copies microL(-1). It is about 100-fold more sensitive than the cell-based infectivity assay. High concentrations of protein and cellular DNA present in test samples have been demonstrated to have no impact on X-MuLV quantification. The X-MuLV clearance during chromatography and filtration procedures determined by this method is highly comparable with that determined by the cell-based infectivity assay. X-MuLV clearance measured by both methods showed that anion exchange chromatography (QSFF) and DV50 viral filtration are robust retroviral removal steps. In addition, combination of the two methods was able to distinguish the viral removal from inactivation by the Protein A chromatography, and fully recognize the viral clearance capacity of this step. This new method offers significant advantages over cell-based infectivity assays. It could be used to substitute cell-based infectivity assays for process validation of viral removal procedures, but not inactivation steps. Its availability should greatly facilitate and reduce the cost of viral clearance evaluations for new biologic product development. PMID- 15334416 TI - Stimulation of methanol degradation in UASB reactors: in situ versus pre-loading cobalt on anaerobic granular sludge. AB - The effect of pre-loading and in situ loading of cobalt onto a cobalt-limited granular sludge on the performance of methanol fed bioreactors was investigated. One upflow anaerobic sludge bed (UASB) reactor was inoculated with cobalt pre loaded sludge (24h; 30 degrees C; 1 mM CoCl2) and a second UASB with unloaded sludge. The UASB reactors (30 degrees C; pH 7) were operated for 77 days at 8 h hydraulic retention time and organic loading rates ranging from 5 to 20 g COD.L reactor(-1).d(-1). Cobalt pre-loading clearly stimulated the methanogenic activity of the sludge with methanol as the substrate, e.g., after 30 days of reactor operation this activity was 5.8 times higher than that of the cobalt unloaded sludge. During the experiment, part of the cobalt leached from the pre loaded sludge, i.e., 54% of the cobalt content was lost during the 77 days of reactor operation. Sequential metal extraction showed that losses mainly occurred from the exchangeable and carbonate fraction and in the sludge remaining cobalt was mainly present in the organic/sulfide fraction of the sludge. In situ loading of cobalt in the unloaded UASB reactor on day 57 by adding 31 microM cobalt to the influent for a 24-h period (16% of the cobalt present in the loaded sludge at day 11) resulted in a 4 time increase of the methanogenic activity of the sludge with methanol as the substrate at the end of the reactor experiment, while the accumulated amount of cobalt in the sludge only amounted to 6% of the cobalt accumulated in the loaded sludge (on day 11). This study showed that both pre loading sludge and in situ loading are adequate for achieving an increased reactor performance of methanol fed UASB reactors operating under cobalt limitation. However, the in situ dosing procedure needs substantially lower amounts of cobalt, while it also gives significantly smaller losses of cobalt with the effluent. PMID- 15334417 TI - Development of an automated water toxicity biosensor using Thiobacillus ferrooxidans for monitoring cyanides in natural water for a water filtering plant. AB - An on-line biosensor consisting of immobilized Thiobacillus ferrooxidans and an oxygen electrode was developed for automated monitoring of acute toxicity in water samples. T. ferrooxidans is an obligatory acidophilic, autotrophic bacterium and derives its energy by the oxidation of ferrous ion, elemental sulfur, and reduced sulfur compounds including metal sulfides. The assay is based on the monitoring of a current increase by addition of toxicoids, which is caused by the inhibition of bacterial respiration and decrease in oxygen consumption. Optimum cell number on the membrane was 5.0 x 10(8) cells. The steady-state current was obtained when concentration of FeSO4 was above 3.6 mM at pH 3. The sensor response of T. ferrooxidans immobilized membrane for 5.0 microM KCN was within an error of 10% for 30 membranes. A linear relationship was obtained at KCN concentration in the range of 0.5-3.0 microM in a flow-type monitoring system. Minimum detectable concentrations of KCN, Na2S, and NaN3 were 0.5, 1.2, and 0.07 microM, respectively. The monitoring system contained two biosensors and these sensors were cleaned with sulfuric acid (pH 1.5) twice a day. This treatment could remove fouling on microbial immobilized membrane by natural water and ferrous precipitation in the flow cell. This flow-type monitoring sensor was operated continuously for 5 months. Also, T. ferrooxidans immobilized membrane can be stored for one month at 4 degrees C when preserved with wet absorbent cotton under argon gas. PMID- 15334418 TI - Bioprocess-centered molecular design (BMD) for the efficient production of an interfacially active peptide. AB - The efficient expression and purification of an interfacially active peptide (mLac21) was achieved by using bioprocess-centered molecular design (BMD), wherein key bioprocess considerations are addressed during the initial molecular biology work. The 21 amino acid mLac21 peptide sequence is derived from the lac repressor protein and is shown to have high affinity for the oil-water interface, causing a substantial reduction in interfacial tension following adsorption. The DNA coding for the peptide sequence was cloned into a modified pET-31(b) vector to permit the expression of mLac21 as a fusion to ketosteroid isomerase (KSI). Rational iterative molecular design, taking into account the need for a scaleable bioprocess flowsheet, led to a simple and efficient bioprocess yielding mLac21 at 86% purity following ion exchange chromatography (and >98% following chromatographic polishing). This case study demonstrates that it is possible to produce acceptably pure peptide for potential commodity applications using common scaleable bioprocess unit operations. Moreover, it is shown that BMD is a powerful strategy that can be deployed to reduce bioseparation complexity. PMID- 15334419 TI - Solid-state fermentation: a continuous process for fungal tannase production. AB - Truly continuous solid-state fermentations with operating times of 2-3 weeks were conducted in a prototype bioreactor for the production of fungal (Penicillium glabrum) tannase from a tannin-containing model substrate. Substantial quantities of the enzyme were synthesized throughout the operating periods and (imperfect) steady-state conditions seemed to be achieved soon after start-up of the fermentations. This demonstrated for the first time the possibility of conducting solid-state fermentations in the continuous mode and with a constant noninoculated feed. The operating variables and fermentation conditions in the bioreactor were sufficiently well predicted for the basic reinoculation concept to succeed. However, an incomplete understanding of the microbial mechanisms, the experimental system, and their interaction indicated the need for more research in this novel area of solid-state fermentation. PMID- 15334420 TI - Costs of workups for the diagnosis of early arthritis: results of a nationwide survey. AB - OBJECTIVE: To evaluate the costs of workups to diagnose early arthritis. METHODS: In 2000, the French Society for Rheumatology conducted a survey of a representative sample of French and Belgian rheumatologists (n = 239). The respondents were asked to consider 2 hypothetical scenarios, 1 describing undifferentiated arthritis and the other more suggestive of rheumatoid arthritis. They were then asked what diagnostic workup they would order. Costs for each study were determined in 2001 euros, according to the French public health system fee schedules. RESULTS: In total, 151 rheumatologists participated in the study (63%). The mean +/- SD diagnostic costs were 406.5 +/- 194.3 euro for the case with no diagnostic clues, and 280.7 +/- 154.3 euro for the case suggestive of early RA. Responses were very heterogeneous. The 2 main sources of expenditure were immunology tests and imaging. Hospital staff physicians tended to order more expensive workups, and costs tended to vary inversely with physician experience. The most important predictor of cost was diagnostic doubt, as estimated by the number of diagnoses proposed by respondents in each case; each additional diagnosis cost an additional 19.1-26.1 euro. CONCLUSION: Diagnostic workups after a first medical visit for early polyarthritis result in substantial direct costs. This observation and the great variability observed in physicians' practices point out the need for consensus on the appropriate workups for these patients. PMID- 15334421 TI - Positron emission tomography in patients with fibromyalgia syndrome and healthy controls. AB - OBJECTIVE: Abnormal brain findings have previously been described in fibromyalgia syndrome (FMS) by single-photon-emission computed tomography. Our goal was to investigate change in regional cerebral glucose metabolism in people with FMS by positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG). METHODS: Twelve patients with FMS and no comorbid psychiatric diagnosis and 7 healthy pain free controls were studied with FDG-PET in a blinded manner. Those with a psychiatric diagnosis were excluded. Brain scans were obtained using a PET scanner. Semiquantitative analysis of regional 18F-FDG uptake was performed in both cortical and subcortical brain structures. RESULTS: In the resting state, there were no significant differences in 18F-FDG uptake between patients and controls for all brain structures measured. CONCLUSION: FDG-PET scan findings in FMS were not significantly different from healthy controls. Normal results in our study may be explained by discordance between regional cerebral blood flow and regional cerebral glucose metabolism. PMID- 15334422 TI - Validity of the childhood health assessment questionnaire is independent of age in juvenile idiopathic arthritis. AB - OBJECTIVE: To determine whether the Childhood Health Assessment Questionnaire (CHAQ) is valid for the comparison of different age subgroups and for longitudinal studies in juvenile idiopathic arthritis (JIA). METHODS: A CHAQ was administered to 306 children with JIA. Rasch analyses were used to compare the difficulty of each of the 30 items of the questionnaire for children of 2 age groups (> or =10 years old and <10 years old). RESULTS: Independent of the physical disability level assessed by the Rasch model, 8 of the 30 items (27%) of the CHAQ were rated significantly different in the 2 age groups. Despite this age related variation in item difficulty, the impact on the CHAQ disability index using its original scoring system remained low (about 0.25 points on a scale of 0 3). CONCLUSION: The difficulty of 8 of 30 items of the CHAQ depends on the respondent's age. Nevertheless, the design of the CHAQ and its scoring system remove most of the expected physical development bias. PMID- 15334423 TI - Factors associated with the use of complementary and alternative medicine in juvenile idiopathic arthritis. AB - OBJECTIVE: To describe the frequency of complementary and alternative medicine (CAM) use in patients with juvenile idiopathic arthritis (JIA) and to explore whether CAM was associated with patient-specific characteristics, parent-specific characteristics, and medical management factors. METHODS: Parents of children with JIA completed questionnaires that addressed the use of CAM, adherence and perceived problems, parental distress, and coping with childhood illness. Clinical variables were abstracted from the patients' medical files. RESULTS: One hundred-eighteen children with JIA, mean +/- SD age of 10.4 +/- 3.9 years and mean +/- SD disease duration of 4.5 +/- 3.5 years, participated in the study. Ever use of CAM was 33.9% and was higher in patients whose parents used CAM (odds ratio [OR] 5.1, 95% confidence interval [95% CI] 1.4-19.5) and among those who considered themselves as "Canadian" as opposed to belonging to a specific ethnic group (OR 10. 7, 95% CI 1.2-99.8). Adherence to conventional treatment was high for both users and nonusers of CAM. CONCLUSION: Use of CAM is common among patients with JIA. CAM use is not related to any decrease in adherence to conventional medical treatment. PMID- 15334424 TI - Frequent mental distress status among adults with arthritis age 45 years and older, 2001. AB - OBJECTIVE: To identify characteristics and behaviors among persons with arthritis through evaluation of self-perceived mental health status. METHODS: Data were analyzed for adults with arthritis age 45 years or older from the 2001 Behavioral Risk Factor Surveillance System, an ongoing, state-based, random-digit-dialed telephone survey of noninstitutionalized adults living in the United States. RESULTS: The prevalence of frequent mental distress (FMD; > or =14 self-reported mentally unhealthy days in the past 30 days) among persons with arthritis was 13.4%. Among persons with arthritis, those with FMD as compared with those without FMD were more likely to be underweight and obese than normal weight; they also were more likely to be insufficiently active or inactive than following recommended physical activity guidelines. In addition, those with arthritis and FMD were more likely to report disability and impaired physical and general health than were those with arthritis but without FMD. CONCLUSION: Physicians should encourage their patients with arthritis and mental distress to participate in educational and behavioral interventions shown to have both physical and psychological benefits. PMID- 15334425 TI - Measuring quality in arthritis care: the Arthritis Foundation's Quality Indicator set for osteoarthritis. AB - OBJECTIVE: To develop a comprehensive set of explicit process measures to assess the quality of health care for osteoarthritis and to describe the scientific evidence that supports each measure. METHODS: Through a comprehensive literature review, we developed potential quality measures and a summary of existing data to support or refute the relationship between the processes of care proposed in the indicators and relevant clinical outcomes. The proposed measures and literature summary were presented to a multidisciplinary panel of experts in arthritis and pain. The panel rated each proposed measure for its validity as a measure of health care quality. RESULTS: Among 22 measures proposed for osteoarthritis, the expert panel rated 14 as valid measures of health care quality. CONCLUSION: Sufficient scientific evidence and expert consensus exist to support a comprehensive set of measures to assess the quality of heath care for osteoarthritis. These measures can be used to gain an understanding of the quality of care for patients with osteoarthritis. PMID- 15334426 TI - Efficacy and safety of four doses of lumiracoxib versus diclofenac in patients with knee or hip primary osteoarthritis: a phase II, four-week, multicenter, randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To compare the efficacy and tolerability of the novel cyclooxygenase 2 selective inhibitor lumiracoxib with placebo and diclofenac in osteoarthritis (OA). METHODS: Adults (n=583) with knee or hip OA were randomized to receive for 4 weeks lumiracoxib 50, 100, or 200 mg twice daily or 400 mg once daily; placebo; or diclofenac 75 mg twice daily. Efficacy assessments included overall joint pain intensity and Western Ontario and McMaster Universities Osteoarthritis Index subscales; tolerability was evaluated by adverse event and physician reporting. RESULTS: All lumiracoxib doses were superior to placebo in relieving pain, improving stiffness, and improving physical function after 4 weeks. At study endpoint, pain relief was comparable among all lumiracoxib dosages and similar to diclofenac. Lumiracoxib tolerability was superior to diclofenac and comparable to placebo. CONCLUSION: Lumiracoxib provides predictable and sustained relief from pain, stiffness, and impaired physical function in OA. Lumiracoxib shows clinically comparable efficacy and superior tolerability to diclofenac. PMID- 15334427 TI - Knee pain and radiographic osteoarthritis interact in the prediction of levels of self-reported disability. AB - OBJECTIVE: To determine predictors of disability depending on whether joint deformity and pain reporting exist independently or concurrently. METHODS: Subjects were 154 volunteers for an osteoarthritis screening examination. Eligible subjects completed questionnaires for physical function, pain, and depressive symptoms; underwent evoked pain testing for tenderness assessment; and had anteroposterior and lateral radiographs taken of both knees. Two blinded rheumatologists scored the images using Kellgren-Lawrence criteria to determine presence of deformity. RESULTS: Subjects were divided into 3 subgroups based on radiographic evidence of deformity and self-reported pain. Disability was greatest when pain and deformity occurred together (F[2,151] = 18.8, P < 0.0001). Self-reported disability in the absence of deformity was predicted by body mass index, pain threshold, and anxiety symptoms; disability was predicted by the number of osteophytes and depressive symptoms when pain and deformity occurred together. CONCLUSION: Self-reported disability in osteoarthritis of the knee is greatest with concurrent pain and joint deformity. When pain and deformity do not cooccur, disability appears to be related to separate factors, including anxiety and pain threshold (e.g., tenderness). PMID- 15334428 TI - Aberrations in the control of quadriceps muscle force in patients with knee osteoarthritis. AB - OBJECTIVE: To characterize the distribution of error in knee joint proprioception, quadriceps force accuracy and steadiness, and muscle strength in patients with knee osteoarthritis (OA). Special attention was paid to eccentric strength. METHODS: We compared knee OA patients (n=20: 15 women, 5 men) with age- and sex-matched, symptom-free adults. Knee pain and mobility were assessed with standard tests. Knee joint proprioception was measured with a repositioning test. Quadriceps force accuracy and steadiness were determined during a force target tracking task. Maximal voluntary quadriceps force was measured during eccentric, isometric, and concentric contractions. RESULTS: OA patients had knee pain, needed 67% more time to complete 4 functional tasks, and produced 82% more proprioception errors (all P < 0.05). About 80% of this error was due to overshooting the target and 68% of the overshooting error occurred at 2 of the 5 least flexed knee joint positions. OA patients had 89% more errors in accurately matching target forces during submaximal quadriceps contractions and in the same tasks, OA patients also produced these forces with 155% more variability (all P < 0.05). OA patients had especially weakened ability to produce maximal voluntary eccentric strength. CONCLUSION: Quadriceps dysfunction in knee OA includes impaired proprioception, especially in the more extended knee joint positions; impaired ability to accurately and steadily control submaximal force; and impaired eccentric strength. These results have implications for designing exercise and rehabilitation programs for patients with knee OA. PMID- 15334429 TI - Burden of caregiving: evidence of objective burden, subjective burden, and quality of life impacts on informal caregivers of patients with rheumatoid arthritis. AB - OBJECTIVE: To improve understanding of the nature and magnitude of the burden of informal care and also to indicate important areas for improving the current ways in which informal care is investigated. METHODS: Information on objective burden (such as care tasks performed and time investment), subjective burden (using the Caregiver Reaction Assessment instrument and a self-rated burden score), and quality of life (using the EuroQoL instrument) were collected in a postal questionnaire of 153 informal caregivers who provide care for rheumatoid arthritis (RA) patients. RESULTS: Caregivers had been caring for the RA patients on average for >11 years, reflecting the chronic nature of RA. They provide a substantial amount of care (27.4 hours per week) and are moderately strained (24.6 on the self-rated burden scale). Caregivers are relatively healthy on average but caregivers of more severe RA patients are relatively unhealthy, which may indicate health losses due to caregiving. CONCLUSION: Informal care can be burdensome in the context of RA. More information may help assist informal caregivers in caring for RA patients and help to avoid health problems and high subjective burden. PMID- 15334430 TI - Impact of fatigue on health-related quality of life in rheumatoid arthritis. AB - OBJECTIVE: To multidimensionally assess fatigue in rheumatoid arthritis (RA) and to evaluate the impact of fatigue on health-related quality of life (HRQOL). METHODS: The study was conducted in 1999 among 490 RA patients with varying disease duration. Fatigue was measured with the Multidimensional Fatigue Inventory (MFI-20) and HRQOL with a validated Dutch version of the RAND 36-Item Health Survey. We evaluated the impact of fatigue on HRQOL by multiple linear regression analyses taking into account RA-related pain and depressive symptoms. RESULTS: Different aspects of fatigue selectively explained different dimensions of HRQOL. The MFI-20 was entered last to the linear regression models, resulting in an additional increase of explained variance of 1% (mental health) to 14% (vitality). CONCLUSION: The multidimensional portrayal of RA-related fatigue can be used to develop intervention strategies targeted to specific aspects of fatigue. Fatigue, supplementary to RA-related pain and depressive symptoms, appears to be a feasible and treatable target in the clinical management of RA to increase HRQOL. PMID- 15334431 TI - Impact of social support on valued activity disability and depressive symptoms in patients with rheumatoid arthritis. AB - OBJECTIVE: To examine the impact of instrumental and emotional support on valued life activity (VLA) disability and depressive symptoms. Instrumental support was expected to affect VLA disability; emotional support was expected to be associated with depressive symptoms and moderate the impact of VLA disability on depressive symptoms. METHODS: Data were collected over 3 years through interviews with the University of California, San Francisco, Rheumatoid Arthritis Panel. Analyses assessed whether instrumental support predicted later VLA disability and whether emotional support predicted both concurrent and later depressive symptoms. RESULTS: Receiving adequate instrumental support was associated with less subsequent VLA disability. Strong associations were noted between both VLA disability and emotional support with concurrent depressive symptoms. No relationship was found between emotional support and later depression. No evidence was found for the hypothesis that emotional support moderated the impact of VLA disability on depressive symptoms. CONCLUSION: Results highlight the need to assess different types of support and their unique impact on critical outcomes. Instrumental support is beneficial to the maintenance of valued activities, a critical factor in the psychological adjustment of individuals living with rheumatoid arthritis. Emotional support has a significant short-term impact on depression, although it may not buffer the impact of VLA disability on future depression. PMID- 15334432 TI - Differences between participants and nonparticipants in an exercise trial for adults with rheumatoid arthritis. AB - OBJECTIVE: To investigate the generalizability of the results of a randomized controlled trial on the effectiveness of long-term, high-intensity exercises in the rheumatoid arthritis patients in training (RAPIT) trial by comparing the characteristics of the participants with the nonparticipants. METHODS: Participants and nonparticipants were mailed questionnaires on sociodemographic characteristics, health status, reasons not to participate, and attitudes toward intensive exercise. RESULTS: The questionnaires from 892 (75%) nonparticipants and 299 (97%) participants were collected. The nonparticipants were slightly older, more often male, and had longer disease duration than the participants. The nonparticipants perceived their disease as more serious, used fewer disease modifying antirheumatic drugs, had a lower level of education, and a more negative attitude toward intensive exercise. CONCLUSION: The results of the RAPIT trial might not be generalizable to the entire target population. To promote participation in long-term, high-intensity exercises, health professionals should more actively discuss the potential benefits of exercise with their RA patients while taking into consideration specific factors related to participation. PMID- 15334433 TI - Pilot clinical trial of dehydroepiandrosterone (DHEA) versus placebo for Sjogren's syndrome. AB - OBJECTIVE: To screen for potential efficacy and assess feasibility and safety of dehydroepiandrosterone (DHEA) as a treatment for Sjogren's syndrome (SS). METHODS: A 24-week randomized, double-blinded, pilot trial of oral DHEA (200 mg/day) versus placebo was conducted. The primary comparison was to a hypothesized 20% placebo response rate. If 14 consecutive subjects on DHEA did not respond, a Phase III trial would be considered futile. A placebo group of 14 subjects was planned to verify placebo response rate and estimate sample size required for a definitive trial. Response criteria required 20% improvement in at least 2 of 3 domains. Analysis of covariance was used to adjust for baseline differences and for stratified randomization. Outcome measures included visual analog scale questionnaires for dry eye and dry mouth symptoms, lissamine green ocular dye staining and Schirmer I tests, stimulated salivary flow, IgG, and erythrocyte sedimentation rate (ESR). RESULTS: Randomization resulted in 14 DHEA and 14 placebo group subjects. At baseline, mean +/- SD for DHEA versus placebo groups were Schirmer I tests 4.5 +/- 4.5 versus 5.4 +/- 6.1 mm/5 minutes; Van Bijsterveld score 5.3 +/- 2.1 versus 5.5 +/- 2.2; unstimulated saliva 0.03 +/- 0.05 versus 0.04 +/- 0.10 ml/minute; IgG 1,699 +/- 749 versus 1,712 +/- 621 g/dl; and ESR 40 +/- 31 versus 44 +/- 28 mm/hour. Apart from changes over the trial in dry mouth symptoms, no significant differences were noted between the DHEA and placebo groups for dry eye symptoms, objective measures of ocular dryness, stimulated salivary flow; IgG, or ESR. Four DHEA and one placebo group patient dropped out because of adverse effects. Although 7 subjects met response criteria in the DHEA group, 5 met the criteria in the placebo group, and there was no significant difference between groups. CONCLUSION: DHEA showed no evidence of efficacy in SS. Without evidence for efficacy, patients with SS should avoid using unregulated DHEA supplements, since long-term adverse consequences of exposure to this hormone are unknown. PMID- 15334434 TI - Mutual support groups in rheumatic diseases: Effects and participants' perceptions. AB - OBJECTIVE: To investigate in a randomized controlled trial the effects of mutual support groups in rheumatic diseases on social network size, social skills, loneliness, daily functioning, and life satisfaction as well as to identify patients' perceptions of the support group. METHODS: Participants were 112 patients with chronic rheumatic disorders affecting the joints. Data were collected with self-report questionnaires and group interviews. RESULTS: Effects have been found on social skills only. More specifically, mutual support groups at postintervention decreased distress in expressing negative feelings toward other people. This effect did not persist at the 6-month followup evaluation, but at that time an increase in frequency in making one's wishes known to others was found. In patients who attended at least 5 of all 10 sessions, an increase in expressing positive feelings toward others was found at followup. Mutual support groups were evaluated positively. CONCLUSION: Mutual support groups are recommended for patients experiencing difficulties in social interactions. PMID- 15334435 TI - Systemic lupus erythematosus in three ethnic groups. XIX. Natural history of the accrual of the American College of Rheumatology criteria prior to the occurrence of criteria diagnosis. AB - OBJECTIVE: To determine how the American College of Rheumatology (ACR) criteria for the classification of systemic lupus erythematosus (SLE) accrue in a multiethnic cohort of SLE patients. METHODS: SLE patients enrolled in a longitudinal study of outcome were analyzed (LUMINA; Lupus in Minorities: Nature versus nurture) for the manner in which ACR criteria manifestations occurred prior to the accrual of 4 of them. Time at which a criterion was said to be present was determined by review of all previously available medical records and interview. Univariable and multivariable Cox proportional hazard models were examined for the association with time to accrual of 4 ACR criteria; results were reported as hazard ratios. RESULTS: There were 103 Texas Hispanic (of Mexican or Central America ancestry) patients, 55 Puerto Rico Hispanics, 176 African Americans, and 137 Caucasians. The mean +/- SD and median (range) time to accrual of 4 ACR criteria were 29.4 +/- 52.0 months and 9.1 (0-328.7) months; time was shortest for the Texas Hispanics (18.4 +/- 42.8 and 5.0 [0-248] months) and longest for the Caucasians (39.9 +/- 59.3 months and 17.7 [0-324.6] months). Arthritis was the most frequent first criterion (34.5%); it was followed by photosensitivity (18.8%). When 2 criteria occurred from the outset, the most frequent combination was arthritis and antinuclear antibody positivity followed by malar rash and photosensitivity. A Cox-regression multivariable model identified Hispanic ethnicity (from Texas) and HLA-DRB1*0301 as predictors of short time to criteria accrual, whereas older age and married/living together were associated with long time to criteria accrual. CONCLUSION: Significant variability in the evolution of ACR criteria manifestations does occur. Texas Hispanics are more likely to have a rapid evolution of criteria manifestations, but several years may elapse before ACR criteria are accrued. PMID- 15334436 TI - Education level and mortality in systemic lupus erythematosus (SLE): evidence of underascertainment of deaths due to SLE in ethnic minorities with low education levels. AB - OBJECTIVE: To determine if socioeconomic status, as measured by education level, is associated with mortality due to systemic lupus erythematosus (SLE), and to determine if these associations differ among ethnic groups. METHODS: Sex- and race-specific mortality rates due to SLE by education level were computed for persons age 25-64 years using US Multiple Causes of Death data from 1994 to 1997. SLE-specific mortality rates were compared with all-cause mortality rates in 1997 to determine if the association between education level and mortality in SLE was similar to that in other causes of death. RESULTS: Among whites, the risk of death due to SLE was significantly higher among those with lower levels of education, and the risk gradient closely paralleled the 1997 all-cause mortality risks by education level. However, in African American women and men and Asian/Pacific Islander women, the risk of death due to SLE was lower among those with lower education levels, contrary to the associations between education level and all-cause mortality in these groups. Comparing the distribution of education levels among deaths due to SLE and all deaths in 1997, persons with lower education levels were underrepresented among deaths due to SLE in African Americans and Asian/Pacific Islanders. CONCLUSION: Among whites, higher education levels are associated with lower mortality due to SLE. These associations were not present in ethnic minorities, likely due to underascertainment of deaths due to SLE in less-well educated persons. This underascertainment may be due to underreporting of SLE on death certificates, but may also represent underdiagnosis of SLE in ethnic minorities with low education levels. PMID- 15334437 TI - Effects of a stress-reduction program on psychological function, pain, and physical function of systemic lupus erythematosus patients: a randomized controlled trial. AB - OBJECTIVE: To assess the effects of a stress-reduction program on pain, psychological function, and physical function in persons with systemic lupus erythematosus (SLE) who experience pain. METHODS: Ninety-two SLE patients were assigned randomly to receive either biofeedback-assisted cognitive-behavioral treatment (BF/CBT), a symptom-monitoring support (SMS) intervention, or usual medical care (UC) alone. RESULTS: BF/CBT participants had significantly greater reductions in pain and psychological dysfunction compared with the SMS group (pain, P = 0.044; psychological functioning, P < 0.001) and the UC group (pain, P = 0.028; psychological functioning, P < 0.001). BF/CBT had significantly greater improvement in perceived physical function compared with UC (P = 0.035), and improvement relative to SMS was marginally significant (P = 0.097). At a 9-month followup evaluation, BF/CBT continued to exhibit relative benefit compared with UC in psychological functioning (P = 0.023). CONCLUSION: This study supports the utility of a brief stress management program for short-term improvement in pain, psychological function, and perceived physical function among persons with SLE who experience pain. PMID- 15334438 TI - Perceptions of, and willingness to consider, total joint arthroplasty in a population-based cohort of individuals with disabling hip and knee arthritis. AB - OBJECTIVE: To examine perceptions of total joint arthroplasty (TJA) and how they relate to willingness to consider TJA. METHODS: A population-based survey in Oxford County, Ontario, Canada identified 1,735 subjects > or =55 years with disabling hip or knee arthritis; 435 English-speaking respondents with no prior TJA and not on a TJA waiting list were invited to participate, and 379 (37.1%) agreed. We assessed demographics; comorbidity; evaluated and perceived arthritis severity; perceived risks, benefits, indications for, and knowledge of TJA; preferred decision-making style; self efficacy; and willingness to consider TJA by questionnaire. RESULTS: Participants' mean age was 67.6 years; 33.5% were willing (definitely or probably) to consider TJA as a treatment option. Willingness was independently associated with younger age (odds ratio [OR] <75 versus > or =75 years 2.42, P = 0.01); worse perceived arthritis severity (OR per unit increase 1.30, P < 0.001); perceiving TJA as appropriate for moderate, controlled joint pain (OR 3.29, P = 0.004); walking limited to <1 block (OR 1.99, P = 0.015); the risk of revision as acceptable (OR 3.73, P < 0.001); and friends as an important health information source (OR 2.49, P = 0.01). CONCLUSIONS: Participants overestimated the pain and disability needed to warrant TJA. These misperceptions were strongly associated with unwillingness to consider TJA and should be addressed at a population level. PMID- 15334439 TI - Prevalence and impact of musculoskeletal disorders of the upper limb in the general population. AB - OBJECTIVE: To determine the prevalence, interrelation, and impact of musculoskeletal disorders of the upper limb in the general population. METHODS: A total of 9,696 randomly selected adults of working age were surveyed in a 2-stage cross-sectional study involving a screening questionnaire and a standardized physical examination in symptomatic subjects. Age- and sex-specific prevalence rates were estimated for several musculoskeletal disorders and for nonspecific pain in the upper limbs. The overlap and impact on daily activities and healthcare utilization were explored. RESULTS: Among 6,038 first-stage responders, 3,152 reported upper limb symptoms and 1,960 were subsequently examined. Of subjects with pain, 44.8% had 1 or more specific soft-tissue disorders. Site-specific prevalence rates were as follows: shoulder tendinitis 4.5% among men and 6.1% among women; adhesive capsulitis 8.2% among men and 10.1% among women; lateral epicondylitis 1.3% among men and 1.1% among women; de Quervain's disease 0.5% among men and 1.3% among women; other tenosynovitis of the hand or wrist, 1.1% among men and 2.2% among women. Specific disorders tended to cluster (P < 0.001) in individuals, with particular overlap at the shoulder. Compared with subjects with nonspecific pain, those with specific disorders more often reported inability to perform everyday tasks (P < 0.05), consultation with a doctor (P < 0.05), and use of prescribed medication (P < 0.05). CONCLUSION: Upper limb pain is common in the general population and is often associated with physical signs suggestive of specific upper-limb disorders. These disorders have a substantial impact on physical function and use of health care. PMID- 15334440 TI - Fever in biopsy-proven giant cell arteritis: clinical implications in a defined population. AB - OBJECTIVE: To assess the frequency and clinical features of biopsy-proven giant cell arteritis (GCA) patients who had fever at the time of diagnosis of the disease, and the relationship between fever, ischemic complications, and the systemic inflammatory response in GCA. METHODS: A retrospective study of biopsy proven GCA patients diagnosed between 1981 and 2001 was performed at the single referral hospital for a well-defined population in the Lugo region of northwest Spain. Patients were considered as having fever if the axillary temperature at the time of admission or during the followup prior to the onset of corticosteroid therapy was > or =38 degrees C. RESULTS: During the period of study, 21 (10%) of the 210 biopsy-proven GCA patients had fever. Two of them fulfilled criteria for fever of unknown origin. Patients with fever had a lower frequency of severe ischemic manifestations than the rest of biopsy-proven GCA patients. They also exhibited a more severe inflammatory disease, with significant abnormality in most laboratory variables, including higher elevation of erythrocyte sedimentation rate, lower values of hemoglobin, and higher proportion of patients with increased alkaline phosphatase. By logistic regression analysis, we observed that patients with fever had an increased risk of developing anemia (odds ratio [OR] 12.24). In contrast, a negative association between severe ischemic manifestations and fever was found (OR 0.41). CONCLUSION: Biopsy-proven GCA patients with fever constitute a subgroup of patients with more severe inflammatory response and less ischemic disease. PMID- 15334441 TI - Wegener's granulomatosis: Possible role of environmental agents in its pathogenesis. AB - OBJECTIVE: To investigate the possibility that environmental agents contribute to the pathogenesis of Wegener's granulomatosis (WG). METHODS: We undertook an extensive search for possible environmental exposures by developing a comprehensive questionnaire that was administered by telephone interview to 53 patients with WG and 2 control groups: one with osteoarthritis and the other with gout. Questions focused on hobbies and vocations, work, home, and allergies. Exact logistic regression was used to calculate odds ratios and 95% confidence intervals after adjusting for potential confounders. After adjusting for age and sex, data are reported for all exposures with odds ratios >2.0 against either control group or for any allergic propensity. RESULTS: Results suggest that mercury and perhaps lead exposure were positively associated with WG as compared with either control group, although the number of patients exposed was small. A prior history of allergy was also associated with WG as compared with either control group. CONCLUSION: We conclude that heavy metal exposure and a prior history of allergy may play a role in the etiopathogenesis of Wegener's granulomatosis. PMID- 15334442 TI - Pulse cyclophosphamide decreases sympathetic postganglionic activity, controls alveolitis, and normalizes vascular tone dysfunction (Raynaud's phenomenon) in a case of early systemic sclerosis. PMID- 15334443 TI - Acute renal failure and interstitial nephritis in a patient treated with rofecoxib: case report and review of the literature. PMID- 15334444 TI - Treatment with statins does not exhibit a clinically relevant corticosteroid sparing effect in patients with giant cell arteritis. PMID- 15334445 TI - Influence of the surgeon's instruction on postoperative joint replacement disability when measured by the Short Form 36: comment on the article by FitzGerald et al. PMID- 15334448 TI - NF-kappaB: Holy Grail for rheumatoid arthritis? PMID- 15334449 TI - Nonmyeloablative allogeneic hematopoietic stem cell transplantation for autoimmune disease. PMID- 15334450 TI - The discovery and development of antiinflammatory drugs. PMID- 15334451 TI - Gout: on the brink of novel therapeutic options for an ancient disease. PMID- 15334452 TI - Biomarkers, surrogate markers, and design of clinical trials of new therapies for systemic lupus erythematosus. PMID- 15334453 TI - Increased levels of C-reactive protein in serum from blood donors before the onset of rheumatoid arthritis. AB - OBJECTIVE: We previously reported that approximately half of the patients with rheumatoid arthritis (RA) have specific serologic abnormalities (elevated serum concentrations of IgM rheumatoid factor and/or anti-cyclic citrullinated peptide antibodies) starting several years before the onset of symptoms. In this study, the presence of serologic signs of inflammation in patients with preclinical RA was investigated with serial measurements of C-reactive protein (CRP). METHODS: Seventy-nine patients (61% female; mean age at onset of symptoms 51 years) who had been blood donors before the onset of RA were identified. Frozen serum samples from each donor were retrieved, together with 1 sample from a control donor matched for age, sex, and date of donation. CRP was measured using a highly sensitive latex-enhanced assay. The dates of donation were categorized into 15 1 year periods preceding the onset of RA symptoms. For each period, the median CRP levels in the patient and control groups were compared using the Mann-Whitney U test. The course of CRP concentrations over time in the patient group was estimated with random coefficient analysis. RESULTS: A median of 13 samples (range 1-51) per patient were available; the earliest donation was made a median of 7.5 years (range 0.4-14.5 years) before the onset of symptoms. A total of 1,078 patient samples and 1,071 control samples were tested. For all 1-year periods, the median CRP concentration was increased in the patient group compared with the control group, but this difference was statistically significant only for the periods 0-1 year, 1-2 years, and 4-5 years before the onset of symptoms. The CRP concentration increased significantly over time in patients with preclinical RA; levels were slightly higher in the group of patients who had serologic abnormalities before the onset of symptoms than in those without such serologic abnormalities. CONCLUSION: After observing specific serologic abnormalities 5 years before the onset of RA symptoms, we now report increased levels of CRP in blood donors in whom RA later developed; these increases were most common within the 2 years before the onset of symptoms. The preclinical increase in CRP levels was observed both in donors with and in those without serologic abnormalities. PMID- 15334454 TI - Regional variation and differential response to therapy for knee synovitis adjacent to the cartilage-pannus junction and suprapatellar pouch in inflammatory arthritis: implications for pathogenesis and treatment. AB - OBJECTIVE: To use magnetic resonance imaging (MRI) to investigate the importance of knee joint synovitis at the cartilage-pannus junction (CPJ) in rheumatoid arthritis (RA) as compared with synovitis at a distant site in the suprapatellar pouch (SPP) and as compared with CPJ synovitis in the spondylarthropathies (SpA), and to assess the relative response of knee joint synovitis to therapy at the CPJ and SPP sites. METHODS: Dynamic contrast-enhanced MRI (DEMRI) of actively involved knee joints in 24 patients (13 with RA and 11 with SpA) was undertaken. The area of synovitis was calculated at the CPJ and SPP regions of interest in patients with RA and in patients with SpA. Differences in CPJ and SPP synovitis were determined using calculated DEMRI parameters which included the initial rate of contrast enhancement (IRE) and the maximal enhancement (ME). Changes in the synovial area at the CPJ and SPP were also measured in 10 patients with early RA, following treatment with disease-modifying antirheumatic drugs (DMARDs) (either methotrexate or leflunomide). RESULTS: In patients with RA or SpA, the area of synovitis was significantly larger immediately adjacent to the CPJ compared with a distant site at the SPP (in RA, mean synovitis area 162 mm2 at the CPJ versus 114 mm2 at the SPP [P = 0.010]; in SpA, mean synovitis area 214 mm2 at the CPJ versus 143 mm2 at the SPP [P = 0.002]), but the differences in the areas of synovitis at these sites were not significant between the RA and SpA patients. The IRE and ME values were also higher at the CPJ compared with the SPP, both in the RA patients (IRE P = 0.054, ME P = 0.018) and in the SpA patients (IRE P = 0.002, ME P = 0.001). A larger reduction in the area of synovitis was seen at the SPP compared with the CPJ following DMARD therapy in the RA patients (mean reduction 35% at the SPP [P = 0.023] and 12% at the CPJ [P not significant]). CONCLUSION: The non-disease-specific variations in synovitis and the differential responses to therapy in RA patients have implications for improving our understanding of CPJ synovitis. The results suggest that the pathophysiologic events at the CPJ reflect common anatomic, immune system, or biomechanical factors that play a role in modulating the severity of arthritis, and these events are not specific to RA since the same process was observed in other arthritides. PMID- 15334455 TI - The rise and decline of nonsteroidal antiinflammatory drug-associated gastropathy in rheumatoid arthritis. AB - OBJECTIVE: Nonsteroidal antiinflammatory drug (NSAID)-associated gastropathy is a major cause of hospitalization and death. This study was undertaken to examine whether recent preventive approaches have been associated with a declining incidence of NSAID gastropathy, and, if so, what measures may have caused the decline. METHODS: We studied 5,598 patients with rheumatoid arthritis (RA) over 31,262 patient-years at 8 sites. We obtained standardized longitudinal information on the patients that had been previously used to establish the incidence of NSAID gastropathy, and also information on patient risk factors and differences in toxicity between NSAIDs. Consecutive patients were followed up with biannual Health Assessment Questionnaires and medical record audits between 1981 and 2000. The major outcome measure was the annual rate of hospitalization involving bleeding, obstruction, or perforation of the gastrointestinal (GI) tract and related conditions. RESULTS: Rates of GI-related hospitalizations rose from 0.6% in 1981 to 1.5% in 1992 (P < 0.001), and then declined to 0.5% in 2000 (P < 0.001). The fitted spline curve fit the data well (R2 = 0.70). The period of rise was mainly associated with increasing patient age and the GI risk propensity score. The period of decline was associated with lower doses of ibuprofen and aspirin, a decline in the use of "more toxic" NSAIDs from 52% to 42% of patients, a rise in the use of "safer" NSAIDs from 19% to 48% of patients, and increasing use of proton-pump inhibitors, but not with change in age, NSAID exposure, or GI risk propensity score. CONCLUSION: The risk of serious NSAID gastropathy has declined by 67% in these cohorts since 1992. We estimate that 24% of this decline was the result of lower doses of NSAIDs, while 18% was associated with the use of proton-pump inhibitors and 14% with the use of less toxic NSAIDs. These declines in the incidence of NSAID gastropathy are likely to continue. PMID- 15334456 TI - Effects of hypoxia on the expression and activity of cyclooxygenase 2 in fibroblast-like synoviocytes: interactions with monocyte-derived soluble mediators. AB - OBJECTIVE: Rheumatoid synovium is characterized by hyperplasia of fibroblast-like (type B) synoviocytes (FLS), infiltration with mononuclear leukocytes, and tissue hypoxia. Although the latter is well documented, it has received little attention in dissection of the biochemical events that mediate the inflammatory lesion in rheumatoid arthritis (RA). Therefore, this study was designed to assess the effect of hypoxia on FLS responses to the monokine interleukin-1beta (IL-1beta) and to monocyte conditioned medium. METHODS: FLS obtained from serial cultures of synovial fluid aspirates were treated with IL-1beta or monocyte conditioned medium, under normoxia and hypoxia. RESULTS: In hypoxia, transcription of cyclooxygenase 2 (COX-2), expression of COX-2 protein, and production of COX-2 derived eicosanoids and matrix metalloproteinase (MMP) activity by FLS were all increased in response to IL-1beta. In contrast to our recent observations concerning monocytes, there was no change in COX-2 message stability and cytosolic phospholipase A2 activity in the FLS under hypoxia. Treatment of monocyte conditioned medium with an IL-1beta blocking antibody showed that most of the effect of the conditioned medium was attributable to IL-1beta. CONCLUSION: The findings suggest that hypoxia is an important factor in aggravating the inflammatory lesion in RA, through increased production of COX-2-derived nociceptive eicosanoids and increased release of tissue-damaging MMPs. PMID- 15334457 TI - Induction of RANKL expression and osteoclast maturation by the binding of fibroblast growth factor 2 to heparan sulfate proteoglycan on rheumatoid synovial fibroblasts. AB - OBJECTIVE: Rheumatoid arthritis (RA) is characterized by progressive joint destruction. The aim of this study was to clarify the relevance of RA synovial fibroblasts (RASFs) and fibroblast growth factor 2 (FGF-2), which is produced abundantly by RASFs, to the osteoclastogenesis and bone resorption in RA. METHODS: Synovial fibroblasts were prepared from the synovial tissues of 10 patients with active RA and 7 patients with osteoarthritis (OA). The expression of RANKL, intercellular adhesion molecule 1 (ICAM-1), FGF receptor 1 (FGFR-1), and heparan sulfate proteoglycan (HSPG) on synovial fibroblasts was measured by FACScan. Osteoclast formation in cocultures of RASFs and peripheral blood mononuclear cells (PBMCs) was evaluated by tartrate-resistant acid phosphatase staining and a pit-formation assay using dentin slices. RESULTS: FGF-2 induced the expression of both RANKL and ICAM-1 on RASFs more so than on OA synovial fibroblasts (OASFs). FGF-2-induced up-regulation of RANKL and ICAM-1 was inhibited by anti-FGF-2 antibody. Although FGFR-1 was equally expressed on RASFs and OASFs, HSPG was highly expressed on RASFs. Up-regulation of RANKL by FGF-2 on RASFs was diminished by the removal of heparan sulfate with heparitinase. Osteoclast formation from PBMCs induced by RASFs was inhibited by the addition of either heparitinase, anti-ICAM-1 antibody, anti-FGF-2 antibody, or osteoprotegerin. FGF-2-induced RANKL on RASFs and osteoclast formation were suppressed by an inhibitor of ERK. CONCLUSION: FGF-2 was transferred to FGFR-1 through binding to HSPG, which is characteristically expressed on RASFs, resulting in RANKL- and ICAM-1-mediated maturation of osteoclasts via ERK activation. Thus, we propose that FGF-2 not only augments the proliferation of RASFs, but also is involved in osteoclast maturation, which leads to bone destruction in RA. PMID- 15334458 TI - In vivo imaging of protease activity in arthritis: a novel approach for monitoring treatment response. AB - OBJECTIVE: Sensitive noninvasive strategies for monitoring treatment response in rheumatoid arthritis (RA) would be valuable for facilitating appropriate therapy and dosing, evaluating clinical outcome, and developing more effective drugs. Because different proteases are highly up-regulated in RA and contribute significantly to joint destruction, in the present study we investigated whether such enzymes are suitable in vivo imaging biomarkers for early evaluation of treatment response in a murine model of RA. METHODS: Using a protease-activated near-infrared fluorescence (NIRF) imaging "smart" probe, we examined the presence and distribution of fluorescence in arthritic joints of mice with collagen induced arthritis by both noninvasive fluorescence imaging and histology. Proteases that target the Lys-Lys cleavage site, including cathepsin B, activate probe fluorescence. Treatment monitoring data were obtained following methotrexate (MTX) therapy. RESULTS: Twenty-four hours after intravenous injection of the protease sensor, affected toes and paws of arthritic mice showed significantly higher fluorescence intensity than did toes and paws of healthy mice. Fluorescence from the protease probe and cathepsin B antibody histologic staining were localized in the vast majority of cells in the inflamed synovium. In arthritic animals treated with MTX (35 mg of MTX/kg 48 hours prior to probe injection), a significantly lower fluorescent signal (inflamed paws 50%, inflamed toes 70%) was observed as compared with untreated arthritic animals. CONCLUSION: Protease-activated NIRF probes are sensitive means of imaging the presence of target enzymes in arthritic joints and can be used for early monitoring of treatment response to antirheumatic drugs such as MTX. PMID- 15334459 TI - Induction of remission of severe and refractory rheumatoid arthritis by allogeneic mixed chimerism. AB - This report describes the first allogeneic hematopoietic stem cell transplantation (HSCT) performed for the indication of rheumatoid arthritis (RA). We used nonmyeloablative allogeneic HSCT to treat a 52-year-old woman who had treatment-refractory RA and a poor prognosis (24 swollen and 38 involved joints). She was treated with fludarabine, cyclophosphamide, CAMPATH-1H, and CD34-selected HSCT (8 million CD34+ donor cells/kg); the donor was the patient's HLA-matched, rheumatoid factor-negative sister. One year post-HSCT, the patient has had no infection except dermatomal varicella-zoster virus infection and no acute or chronic graft-versus-host disease (GVHD). Her RA has remained in remission with no immunosuppressive or immunomodulatory medications. The patient is a mixed chimera, with 55% donor T (CD3+) cells and 70% donor myeloid (CD33+) cells. This is the first published report of allogeneic HSCT performed for the indication of RA. Mixed chimerism has resulted in marked amelioration of RA, without GVHD. PMID- 15334460 TI - A new marker for osteoarthritis: cross-sectional and longitudinal approach. AB - OBJECTIVE: To investigate the association between urinary concentrations of C telopeptide fragments of type II collagen (CTX-II) and the prevalence and progression of radiographic osteoarthritis (OA) of the knee and hip. METHODS: The study population consisted of a sample of 1,235 men and women ages > or =55 years who were enrolled in the Rotterdam Study (a population-based cohort study) and who were followed up for a mean of 6.6 years. Prevalent radiographic OA was defined as a Kellgren/Lawrence score > or =2; progression of radiographic OA was defined as a decrease in joint space width. RESULTS: Subjects with a CTX-II level in the highest quartile had a 4.2-fold increased risk of having radiographic OA of the knee (95% confidence interval [95% CI] 2.5-7.0) and of the hip (95% CI 2.2 7.8) compared with subjects with a CTX-II level in the lowest quartile. We observed a substantially stronger association between CTX-II levels and radiographic OA for subjects with hip pain (odds ratio [OR] 20.4, 95% CI 2.3 185.2) than for those without hip pain (OR 3.0, 95% CI 1.5-6.0). Subjects with a CTX-II level in the highest quartile had a 6.0-fold increased risk for progression of radiographic OA at the knee (95% CI 1.2-30.8) and an 8.4-fold increased risk for progression of radiographic OA at the hip (95% CI 1.0-72.9). All of these associations were found to be independent of known risk factors for OA, such as age, sex, and body mass index. CONCLUSION: This study shows that CTX II is associated with both the prevalence and the progression of radiographic OA at the knee and hip. Importantly, this association is independent of known clinical risk factors for OA and seems stronger in subjects with joint pain. PMID- 15334461 TI - Suggestion of nonlinear or phasic progression of knee osteoarthritis based on measurements of serum cartilage oligomeric matrix protein levels over five years. AB - OBJECTIVE: In many patients with knee osteoarthritis (OA), the disease progresses, and there is loss of cartilage; in others, the disease stabilizes with time. Previous studies have demonstrated that concentrations of serum proteins that reflect joint tissue metabolism can identify knees that will deteriorate, leading to the suggestion that OA disease activity is phasic or cyclical. The aim of the current study was to determine whether longitudinal measurements of one such protein, serum cartilage oligomeric matrix protein (COMP), are related to disease outcome over a 5-year period. METHODS: Serum COMP levels were measured by enzyme-linked immunosorbent assay at study entry and every 6 months thereafter in 115 patients with knee pain and OA of mainly the tibiofemoral joint. Cartilage loss was determined from knee radiographs taken at entry and at 24, 36, and 60 months. Disease progression was defined as either a reduction in the tibiofemoral joint space width by at least 2 mm or total knee replacement (TKR) in either knee at followup. COMP concentrations at baseline and the area under the curve (AUC) of measurements obtained over 5 years were compared between progressors and nonprogressors by Student's 2-tailed t-test. The patterns and probability of progression according to TKR or > or =2 mm of narrowing of the tibiofemoral joint space were analyzed by logistic regression models. RESULTS: The mean +/- SD ages of the progressors and nonprogressors were 64.2 +/- 7.8 years and 63.3 +/- 10.6 years, respectively, and the proportion of females was 51% and 56%, respectively. Of the 37 patients whose OA progressed (22 by TKR and 15 by > or =2-mm reduction in tibiofemoral joint space), 13 lost cartilage during the first 2 years, and 18 lost cartilage during the last 2 years. The mean +/- SD serum COMP concentration at baseline was significantly higher in the progressors compared with the nonprogressors (14.12 +/- 3.39 units/liter versus 12.62 +/- 3.25 units/liter; P < 0.036). Serum COMP levels rose significantly after TKR; however, after allowing for the effect of TKR, the AUC/month was significantly higher in the progressors compared with the nonprogressors (12.52 +/- 2.71 versus 10.82 +/- 2.71; P < 0.003). Serum COMP concentrations were higher during periods of radiographic progression and identified periods of progression that were nonlinear. Logistic regression analysis showed that on average, a 1-unit increase in serum COMP levels increased the probability of radiographic progression by 15%. CONCLUSION: The data suggest that serum COMP is related to progressive joint damage in knee OA. The patterns of progression for the early and late progressors are consistent with the hypothesis that knee OA progression is episodic or phasic. Large between-subject variation precludes the use of individual values to predict progression with confidence. However, sequential measurements of serum COMP levels may identify patients whose OA is likely to progress over the next year or two. PMID- 15334462 TI - A genome scan for joint-specific hand osteoarthritis susceptibility: The Framingham Study. AB - OBJECTIVE: Studies investigating hand osteoarthritis (OA) as a single entity have not shown strong linkage of the disease with any chromosomal sites. We undertook this study to test our hypothesis that phenotypes of hand OA may show stronger linkage than has been shown for overall hand OA. METHODS: We performed a factor analysis on measures of hand OA to determine patterns of disease. Using the joint regions identified by this analysis, we performed a genome-wide linkage analysis for OA susceptibility loci using 426 original cohort members and 790 offspring cohort members in 267 pedigrees. Radiographic OA features evaluated included the Kellgren/Lawrence score, osteophytes, and joint space narrowing. Prior to linkage analysis, standardized residuals were computed from regression analysis of each phenotype on age. This was performed separately for each sex and cohort. The variance component model (GeneHunter) was then applied to the normalized scores of the residuals of both sexes and cohorts. RESULTS: There was evidence suggestive of linkage (logarithm of odds [LOD] score >1.5) at 16 sites. Four of these sites had LOD scores >3.0. Two of these sites (identified in the full sample) included a linkage region for OA of the distal interphalangeal (DIP) joint on chromosome 7 (155 cM; LOD score 3.06) and a linkage region for OA of the first carpometacarpal (CMC) joint on chromosome 15 (81 cM; LOD score 6.25). The other 2 sites (identified in women) included a linkage region for OA of the DIP joint on chromosome 1 (202 cM; LOD score 3.03) and a linkage region for OA of the first CMC joint on chromosome 20 (4 cM; LOD score 3.74). CONCLUSION: These data suggest that several chromosomes contain hand OA susceptibility genes and that a joint-specific approach may be more rewarding than a global approach to the genetics of hand OA. Further investigation of these regions is warranted using finer maps and other populations. PMID- 15334463 TI - Association study of candidate genes for the prevalence and progression of knee osteoarthritis. AB - OBJECTIVE: Osteoarthritis (OA), characterized by late-onset degeneration of articular cartilage, is recognized to have a genetic component. We examined the role of 26 single-nucleotide polymorphisms (SNPs) from 24 candidate genes in OA susceptibility and progression. METHODS: We compared human complementary DNA libraries from OA-affected and normal cartilage and synovium and selected 22 genes in addition to the estrogen receptor alpha and vitamin D receptor genes. Based on the availability of polymorphisms, we proceeded to test whether genetic variation at those genes affected susceptibility to or progression of radiographic knee OA over a 10-year period in 749 women (mean age 64 years) from the longitudinal Chingford Study. RESULTS: After adjusting for age and body mass index, we observed significant associations at ADAM12, BMP2, CD36, COX2, and NCOR2 with 3 OA susceptibility traits (presence/absence of joint space narrowing [JSN], presence/absence of osteophytes, and Kellgren/Lawrence [K/L] score). For the OA progression traits (change over 10 years in the K/L score, osteophyte grade, and JSN grade), we found significant associations with ADAM12, CILP, OPG, and TNA. Overall, we observed 15 associations with nominal significance (P < 0.05) and, by permutation analysis, found that such a number would be observed by chance only 3.8% of the time. Although these tests require replication, the stronger genetic associations observed are unlikely to be attributable simply to multiple comparisons. CONCLUSION: Our results suggest that OA severity and progression have a multigenic and feature-specific nature. These findings should encourage the development of genetic diagnostics for OA progression based on multiple SNPs and help unravel some of the complex disease mechanisms in OA. PMID- 15334464 TI - Pitfalls in the accurate measurement of joint space narrowing in semiflexed, anteroposterior radiographic imaging of the knee. AB - OBJECTIVE: Computerized measurement of changes in joint space width (JSW) on serial radiographs of the knee in the semiflexed, anteroposterior (SF-AP) view has been used recently as a primary outcome measure in clinical trials of disease modifying osteoarthritis drugs (DMOADs). In the use of fluoroscopy to achieve reproducible alignment of the medial tibial plateau and x-ray beam, the SF-AP radiographic protocol affords greater sensitivity in the detection of joint space narrowing (JSN) than that achieved by conventional radiographic positioning techniques. However, the utility of the SF-AP view is compromised by the variation in x-ray penetration in each examination, which may confound the correction of the automated measurement of JSW for the radiographic magnification inherent in an AP view of the knee. A recent DMOAD trial using the SF-AP protocol showed an improbable increase in JSW of > or =0.50 mm (i.e., greater than the measurement error). The present report provides an analysis of this problem, and the study aim was to demonstrate that substitution of the automated estimates of JSW with precise manual measurements can markedly reduce the problem attributable to radiographic magnification. METHODS: SF-AP radiographs were obtained at baseline and at 16 months and 30 months thereafter from subjects enrolled in a 6 center DMOAD trial. For each examination, a 6.35-mm steel ball was affixed to the skin over the head of the fibula to permit estimation of the percentage of radiographic magnification (%Mag) and correction of JSW measurements. Measurements of the minimum interbone distance (IBD) in the medial tibiofemoral compartment and the %Mag were obtained by an automated method (edge detection) and manually. Combinations of automated and manual measurements of the IBD and %Mag in estimates of magnification-corrected JSW were compared with respect to their reproducibility, agreement, and sensitivity to JSN. RESULTS: With fully automated measurements, variations in x-ray penetration in analog radiographs and edge enhancement in digital radiographs resulted in the computer "seeing" a metal ball whose diameter was artifactually reduced, resulting in an inflated measurement of JSW. Use of manual measurement of the IBD and %Mag largely eliminated these problems and reduced, from 16% to 2%, the frequency of knees exhibiting an increase in JSW > or =0.50 mm. In 14 of the 15 knees in which a significant increase in JSW was noted with the manual method, this increase in JSW could be explained by the development of significant lateral compartment narrowing during the study or poor alignment of the medial plateau. CONCLUSION: Although automated and manual methods of JSW measurement of the knee in the SF-AP view possess comparable intrareader reproducibility, the manual method is less susceptible to technical factors that affect the correction of raw JSW estimates for radiographic magnification. Until we can identify practical, effective solutions to these technical problems, use of any radiographic protocol involving AP imaging of the knee in a DMOAD trial must be viewed with caution. PMID- 15334465 TI - Cartilage atrophy in the knees of patients after seven weeks of partial load bearing. AB - OBJECTIVE: It is currently unknown whether human cartilage properties change during short periods of partial load bearing. We used a post-ankle fracture model to explore whether changes in cartilage morphology occur in the knee under conditions of partial load bearing. METHODS: The knees of 20 patients with Weber type B and type C fractures were examined using magnetic resonance imaging. The first scan was obtained shortly (mean +/- SD 3.2 +/- 3.0 days) after the injury, and a second scan was obtained 7 weeks later (mean +/- SD 50.7 +/- 5.5 days). The morphology (mean and maximum thickness, volume, and surface area) of the patellar, tibial, and femoral cartilage was determined from coronal and axial magnetic resonance images (fat-suppressed gradient-echo). RESULTS: Between week 0 and week 7, the cross-sectional area of the quadriceps muscle was reduced by 11% (P< 0.001). Changes in the mean (+/-SD) cartilage thickness ranged from -2.9 +/- 3.2% in the patella to -6.6 +/- 4.9% in the medial tibia. No significant change in cartilage morphology of the contralateral knee was observed. CONCLUSION: Results of this study demonstrate that in a post-ankle fracture model of partial load bearing, cartilage morphology in all knee compartments is subject to significant change. Changes in the femorotibial joint exceeded those in the patella, whereas no change was observed in the contralateral knee. These findings raise the question of whether cartilage is mechanically less competent and particularly vulnerable after states of partial or complete immobilization. PMID- 15334466 TI - Differential gene expression and regulation of the bone morphogenetic protein antagonists follistatin and gremlin in normal and osteoarthritic human chondrocytes and synovial fibroblasts. AB - OBJECTIVE: To compare gene expression in normal and osteoarthritic (OA) human chondrocytes using microarray technology. Of the novel genes identified, we selected follistatin, a bone morphogenetic protein (BMP) antagonist, and investigated its expression/regulation as well as that of 3 other antagonists, gremlin, chordin, and noggin, in normal and OA chondrocytes and synovial fibroblasts. METHODS: Basal and induced gene expression were determined using real-time polymerase chain reaction. Gene regulation was monitored following treatment with inflammatory, antiinflammatory, growth, and developmental factors. Follistatin protein production was measured using a specific enzyme-linked immunosorbent assay, and localization of follistatin and gremlin in cartilage was determined by immunohistochemical analysis. RESULTS: All BMP antagonists except noggin were expressed in chondrocytes and synovial fibroblasts. Follistatin and gremlin were significantly up-regulated in OA chondrocytes but not in OA synovial fibroblasts. Chordin was weakly expressed in normal and OA cells. Production of follistatin protein paralleled the gene expression pattern. Follistatin and gremlin were expressed preferentially by the chondrocytes at the superficial layers of cartilage. Tumor necrosis factor alpha and interferon-gamma significantly stimulated follistatin expression but down-regulated expression of gremlin. Interleukin-1beta (IL-1beta) had no effect on follistatin but reduced gremlin expression. Conversely, BMP-2 and BMP-4 significantly stimulated expression of gremlin but down-regulated that of follistatin. IL-13, dexamethasone, transforming growth factor beta1, basic fibroblast growth factor, platelet-derived growth factor type BB, and endothelial cell growth factor down regulated the expression of both antagonists. CONCLUSION: This study is the first to show the possible involvement of follistatin and gremlin in OA pathophysiology. The increased activin/BMP-binding activities of these antagonists could affect tissue remodeling. The data suggest that follistatin and gremlin might appear at different stages during the OA process, making them interesting targets for the treatment of this disease. PMID- 15334467 TI - Effects of dehydroepiandrosterone on articular cartilage during the development of osteoarthritis. AB - OBJECTIVE: To investigate the in vivo effects of dehydroepiandrosterone (DHEA) on knee joints during the development of experimentally induced osteoarthritis (OA). METHODS: Twenty-two mature NZW rabbits underwent bilateral anterior cruciate ligament transection (ACLT) and received 0.3-ml intraarticular injections of DHEA (at a concentration of 100 microM in phosphate buffered saline) and control solution in the right and left knees, respectively, beginning 4 weeks after ACLT and continuing once weekly for 5 weeks. All animals were killed 9 weeks after surgery, and the knee joints were assessed by gross morphologic, histologic, histomorphometric, and biochemical methods. RESULTS: Gross morphologic inspection following India ink application showed that the right femoral condyles, which received DHEA, demonstrated less severe cartilage damage than did the contralateral condyles. The thickness, area, and roughness of the DHEA-treated femoral condyles provided evidence of a cartilage-protecting effect of DHEA following ACLT. These results were supported by gene expression analysis. Messenger RNA expression of a proinflammatory cytokine, interleukin-1beta, and catabolic enzymes, matrix metalloproteinases 1 and 3, was reduced in the cartilage of the DHEA-treated knee joints, and expression of tissue inhibitor of metalloproteinase 1 was increased. CONCLUSION: Results of the present study demonstrate a cartilage-protecting effect of DHEA during the development of OA following ACLT in a rabbit model. PMID- 15334468 TI - Induction of interleukin-1 in articular cartilage by explantation and cutting. AB - OBJECTIVE: To investigate the effect of explantation and fine cutting of articular cartilage upon intracellular inflammatory signaling pathways and expression of interleukin-1 (IL-1). METHODS: Cartilage from porcine metacarpophalangeal joints was cultured in serum-free medium. Tissue extracts were examined for ERK activation by phosphorylated-Western blotting, for JNK and p38 MAPK activity by kinase assay, and for IkappaBalpha. IL-1alpha and IL-1beta messenger RNA (mRNA) was measured by reverse transcriptase-polymerase chain reaction. IL-1 activity was measured by the induction of serum amyloid A protein in cultured chondrocytes. RESULTS: All 3 MAPKs (p38, JNK, and ERK) were rapidly activated upon dissection and explantation of the cartilage. IL-1alpha and IL 1beta mRNA was also induced: the speed and magnitude of induction were increased if the explants had been finely cut. IL-1 activity that could be inhibited by IL 1 receptor antagonist or antibodies to IL-1alpha was found in extracts of explants cultured for 20 hours or lysates of cells isolated from them. This activity was likely due to intracellular proIL-1alpha that was not secreted. ProIL-1beta would not be detected because it is biologically inactive. The mechanism of inflammatory signaling pathway activation underlying the induction of IL-1 is unknown. CONCLUSION: Explantation and cutting of articular cartilage activates intracellular inflammatory signaling pathways and induces expression of mRNA for IL-1alpha and IL-1beta. Biologically active IL-1alpha protein was detectable in cartilage lysates and was probably intracellular proIL-1alpha. We were unable to show that IL-1 was secreted by chondrocytes. PMID- 15334469 TI - Characterization of and osteoarthritis susceptibility in ADAMTS-4-knockout mice. AB - OBJECTIVE: To determine the importance of the enzymatic activity of ADAMTS-4 in normal growth and development and to evaluate the role of ADAMTS-4 in the progression of osteoarthritis (OA). METHODS: We generated catalytic domain deleted ADAMTS-4-transgenic mice and performed extensive gross and histologic analyses of various organs. The mice were challenged by surgical induction of joint instability leading to OA, to determine the importance of the enzymatic activity of ADAMTS-4 in the progression of the disease. The response of wild-type (WT) and ADAMTS-4-knockout (ADAMTS-4-KO) articular cartilage to interleukin-1 and retinoic acid challenge in vitro was also evaluated. RESULTS: ADAMTS-4-KO mice up to 1 year of age exhibited no gross or histologic abnormalities in 36 tissue sites examined. Despite evidence of ADAMTS-4 expression and activity in growth plates of WT mice, catalytic silencing of this proteinase caused no abnormalities in skeletal development, growth, or remodeling. There was no effect of ADAMTS-4 knockout on the progression or severity of OA 4 weeks or 8 weeks after surgical induction of joint instability. Enzymatic cleavage of aggrecan at the TEGE(373 374)ARGS site was clearly evident after exposure of articular cartilage from ADAMTS-4-KO mice to inflammatory cytokines. CONCLUSION: Although expression of the ADAMTS-4 gene has been found in many tissues throughout the body, deletion of enzymatic activity did not appear to have any effect on normal growth and physiology. Our study provides evidence that ADAMTS-4 is the primary aggrecanase in murine growth plates; however, deletion of its enzymatic activity did not affect normal long bone remodeling. Our results also lead to the hypothesis that, in the mouse, ADAMTS-4 is not the primary enzyme responsible for aggrecan degradation at the TEGE(373-374)ARGS site. The elucidation of the relative importance of ADAMTS-4 in the pathologic process of human OA will require examination of human OA tissues and evidence of disease modification in patients following therapeutic intervention. PMID- 15334470 TI - Predictors and outcome of renal flares after successful cyclophosphamide treatment for diffuse proliferative lupus glomerulonephritis. AB - OBJECTIVE: To study the incidence, predictors, and outcome of renal flares after successful cyclophosphamide (CYC) treatment for diffuse proliferative glomerulonephritis (DPGN) in patients with systemic lupus erythematosus (SLE). METHODS: Between 1988 and 2001, patients with biopsy-proven SLE DPGN who were treated initially with prednisone and CYC were studied. Those who responded to CYC were followed up for the occurrence of renal flares. The cumulative risk, predictors, and outcome of renal flares were evaluated. RESULTS: We studied 189 patients (167 women; and 22 men) with SLE DPGN. All were initially treated with prednisone and CYC (49% orally; 51% by intravenous pulse). At the last dose of CYC, 103 patients (55%) and 52 patients (28%) had achieved complete and partial renal responses, respectively. Azathioprine (AZA) was given as maintenance therapy in 117 patients (75%). After a mean followup of 96.5 months, 59 patients (38%) experienced renal flares (42% nephritic; 58% proteinuric). The median time to relapse was 32 months. The cumulative risk of renal flare was 28% at 36 months and 44% at 60 months. Independent predictors of nephritic flares were persistently low C3 levels after CYC treatment and absence of AZA maintenance therapy. At the last clinic visit, 16 patients (10.3%) had developed doubling of the serum creatinine level (cumulative risk of creatinine doubling 7.4% at 5 years after renal biopsy and 14.3% at 10 years). Ten patients (6.5%) developed end-stage renal disease (ESRD). Renal survival rates at 5 and 10 years were 94.9% and 87.5%, respectively. Increasing histologic chronicity scores, failure to achieve complete response, persistent hypertension after CYC treatment, and nephritic renal flares were unfavorable factors for doubling of the serum creatinine level and for ESRD by univariate analysis. The occurrence of nephritic flares was the only predictor of creatinine doubling by Cox regression analysis. CONCLUSION: In patients with SLE DPGN, renal flares are common despite initial responses to CYC. Nephritic renal flares are associated with a decline in renal function. Maintenance therapy with AZA reduces, but does not completely prevent, renal flares. More effective maintenance treatment for SLE DPGN after an initial response to CYC should be evaluated. PMID- 15334471 TI - Antiphospholipid syndrome nephropathy in patients with systemic lupus erythematosus and antiphospholipid antibodies: prevalence, clinical associations, and long-term outcome. AB - OBJECTIVE: To evaluate the prevalence, clinical associations, and outcome of antiphospholipid syndrome (APS) nephropathy in patients with systemic lupus erythematosus (SLE) and antiphospholipid antibodies (aPL) and in SLE patients without aPL. METHODS: Kidney biopsy specimens obtained from 81 patients with aPL (18 of whom had APS) and 70 patients without aPL were retrospectively examined for the presence of APS nephropathy. Clinical and serologic data obtained at the time of kidney biopsy and during a mean followup of 7 years were recorded. In cases for which serial kidney biopsy specimens were available, the evolution of APS nephropathy was examined. RESULTS: APS nephropathy existed in 39.5% of patients with aPL, compared with only 4.3% of patients without aPL. APS nephropathy was associated with both lupus anticoagulant and anticardiolipin antibodies. Among aPL-positive SLE patients, APS nephropathy was found in two thirds of those with APS and in one-third of those without APS. A strong association between APS nephropathy and the presence of arterial thrombosis and livedo reticularis was noted. Patients with APS nephropathy had a higher frequency of hypertension and elevated serum creatinine levels at the time of kidney biopsy but did not have a higher frequency of renal insufficiency, end stage renal disease, or death at the end of followup. Serial kidney biopsy specimens were available from 11 patients and showed progression of APS nephropathy lesions. During followup, manifestations of APS (especially arterial thromboses) developed more frequently in the SLE/non-APS patients with APS nephropathy than in those without APS nephropathy. CONCLUSION: Among patients with SLE, APS nephropathy occurs almost exclusively in those with aPL, suggesting an important role of aPL in the pathogenesis of APS nephropathy. Patients with APS nephropathy develop hypertension, raised serum creatinine levels, and progression of histologic lesions, all of which are associated with a poor renal outcome. Manifestations of APS also tend to develop in these patients. APS nephropathy should be included in the APS classification criteria, and the use of appropriate anticoagulant therapy should be tested. PMID- 15334472 TI - B cell depletion as a novel treatment for systemic lupus erythematosus: a phase I/II dose-escalation trial of rituximab. AB - OBJECTIVE: Safer and more effective therapies are needed for the treatment of systemic lupus erythematosus (SLE). B lymphocytes have been shown to play fundamental pathogenic roles in SLE, and therefore, elimination of B cells with the use of rituximab may represent a new therapy for SLE. METHODS: A phase I/II dose-escalation trial of rituximab added to ongoing therapy in SLE was conducted. Rituximab was administered as a single infusion of 100 mg/m2 (low dose), a single infusion of 375 mg/m2 (intermediate dose), or as 4 infusions (1 week apart) of 375 mg/m2 (high dose). CD19+ lymphocytes were measured to determine the effectiveness of B cell depletion. The Systemic Lupus Activity Measure (SLAM) score was used as the primary outcome for clinical efficacy. RESULTS: Rituximab was well tolerated in this patient population, with most experiencing no significant adverse effects. Only 3 serious adverse events, which were thought to be unrelated to rituximab administration, were noted. A majority of patients (11 of 17) had profound B cell depletion (to <5 CD19+ B cells/microl). In these patients, the SLAM score was significantly improved at 2 and 3 months compared with baseline (P = 0.0016 and P = 0.0022, respectively, by paired t-test). This improvement persisted for 12 months, despite the absence of a significant change in anti-double-stranded DNA antibody and complement levels. Six patients developed human antichimeric antibodies (HACAs) at a level > or =100 ng/ml. These HACA titers were associated with African American ancestry, higher baseline SLAM scores, reduced B cell depletion, and lower levels of rituximab at 2 months after initial infusion. CONCLUSION: Rituximab therapy appears to be safe for the treatment of SLE and holds significant therapeutic promise, at least for the majority of patients experiencing profound B cell depletion. Based on these results, controlled trials of rituximab appear to be warranted. PMID- 15334473 TI - Association of PDCD1 with susceptibility to systemic lupus erythematosus: evidence of population-specific effects. AB - OBJECTIVE: The A allele of the PD1.3 single-nucleotide polymorphism (SNP) on the programmed cell death gene PDCD1 was markedly more frequent in patients with systemic lupus erythematosus (SLE) than in unaffected controls in a recent study involving large sets of Swedish, European American, and Mexican families. This study sought to determine the role of PDCD1 in susceptibility to SLE in the Spanish population. METHODS: Seven PDCD1 SNPs were studied in 518 SLE patients and 800 healthy control subjects who had been recruited in 5 distant towns spanning continental Spain. Patients and controls were of Spanish ancestry. The diagnosis of SLE was in accordance with the American College of Rheumatology updated classification criteria. RESULTS: The A allele of the PD1.3 polymorphism was significantly less frequent in Spanish female patients with SLE than in Spanish female controls (9.0% versus 13.0%, odds ratio 0.67, 95% confidence interval 0.50-0.89). This difference was consistent across the 5 sets of samples grouped by town of recruitment. The other PDCD1 SNPs were not associated with SLE susceptibility. The haplotype structure of PDCD1 in the Spanish controls was different from that reported in other healthy control populations. CONCLUSION: Our results confirm the association of PDCD1 with susceptibility to SLE, but the findings show a lack of involvement of the PD1.3 SNP, which is contrary to the role of the PD1.3 A allele observed previously. These contradictory results probably reflect population differences in the haplotype structure of the PDCD1 locus. More research focusing on new polymorphisms and identifying associations in other populations will be needed to clarify the role of PDCD1 in SLE susceptibility. PMID- 15334474 TI - Genetic association of cutaneous neonatal lupus with HLA class II and tumor necrosis factor alpha: implications for pathogenesis. AB - OBJECTIVE: Cutaneous neonatal lupus resembles subacute cutaneous lupus erythematosus (SCLE), and photosensitivity is a common symptom. Tumor necrosis factor alpha (TNFalpha) release by ultraviolet light-exposed keratinocytes may be exaggerated in SCLE patients who have the haplotype TNFalpha -308A;DRB1*03. Accordingly, this study was undertaken to seek genetic and histologic evidence for a role of TNFalpha in the pathogenesis of cutaneous neonatal lupus. METHODS: DNA was isolated from 83 children (22 with rash, 35 with congenital heart block [CHB], 26 unaffected siblings) and 58 mothers from the Research Registry for Neonatal Lupus. RESULTS: The -308A allele (associated with higher TNFalpha production), HLA-DRQB1*02, and HLA-DRB1*03 were each present in the majority of children with rash (64%, 68%, and 64%, respectively). The frequency of all 3 6p alleles occurring together in 1 individual was greater in children with rash than in children who had either CHB or no manifestation of neonatal lupus (59% versus 30%; P = 0.02). This association with neonatal lupus rash was equivalent to published findings in a cohort of patients with SCLE, but significantly greater than the association in patients with discoid lupus erythematosus. Prominent TNFalpha staining in the epidermis was observed in lesional skin from 3 children with rash, but not in skin from a healthy neonate. CONCLUSION: Taken together, the finding of a genetic predisposition to generate increased levels of TNFalpha following tissue injury and the histologic demonstration of TNFalpha in the target organ support the notion that this inflammatory cytokine plays a role in the pathogenesis of cutaneous neonatal lupus. Furthermore, the results of these studies provide evidence of a biologic link between neonatal lupus and the rash of SCLE. PMID- 15334475 TI - The V(lambda)-J(lambda) repertoire of patients with systemic lupus erythematosus manifests characteristics of the natural antibody repertoire. AB - OBJECTIVE: To understand in detail the mechanisms of autoantibody production in patients with systemic lupus erythematosus (SLE), we performed a comprehensive analysis of the normal human immunoglobulin light chain V(lambda) repertoire and compared it with the V(lambda) repertoire in SLE patients. METHODS: The SLE V(lambda) repertoire of B cells obtained from 3 SLE patients was analyzed and compared in detail with the V(lambda) repertoire of IgM+ B cells obtained from 3 human fetal spleens and IgM+,CD5+ B cells obtained from 2 normal adults. Conventional IgM+,CD5- B cells obtained from normal adults were used as controls. V(lambda)-J(lambda) rearrangements were amplified from the genomic DNA of individual B cells by polymerase chain reaction. RESULTS: The expressed V(lambda) repertoire of SLE patients contained several similarities with the expressed repertoire of the fetus and the adult CD5+ B cells. The V(lambda) genes 3L and 1G were overexpressed in the fetus, the adult CD5+ B cells, and the patients with SLE. The selection for rearrangements with restricted junctional diversity by utilization of homology-mediated joining, together with diminished N nucleotide addition, was a prominent feature of fetal, adult CD5+, and SLE B cell repertoires. Furthermore, profound expansion of V(lambda) clones with identical third complementarity-determining regions was observed in the adult CD5+, fetal, and SLE B cell repertoires. Notably, significant numbers of expanded adult CD5+ B cells, fetal, and SLE V(lambda) clones utilized homology-mediated joining at the V(lambda)-J(lambda) junctions. CONCLUSION: These data demonstrate that the SLE V(lambda)-J(lambda) repertoire manifests characteristics of normal adult IgM+,CD5+ and fetal B cell populations that are known to be enriched for the production of natural autoantibodies. PMID- 15334476 TI - Detection of anti-type 3 muscarinic acetylcholine receptor autoantibodies in the sera of Sjogren's syndrome patients by use of a transfected cell line assay. AB - OBJECTIVE: Sjogren's syndrome (SS) is an autoimmune disease affecting primarily the salivary and lacrimal glands, leading to dry mouth and dry eyes. Recent studies have suggested that autoantibodies reactive with the type 3 muscarinic acetylcholine receptors (M3Rs) expressed on salivary and lacrimal gland cells may be highly specific for SS. To test this hypothesis, we constructed a cell line expressing the human M3R gene in order to screen for anti-M3R autoantibodies in sera from SS patients. METHODS: Complementary DNA encoding the open-reading frame (ORF) of the human M3R gene was amplified, ligated into the pcDNA5/FRT/V5-His TOPO TA vector, and then used to transform Escherichia coli bacteria. Plasmid DNA containing the M3R ORF with the correct orientation was transfected into Flp-In Chinese hamster ovary (CHO) cells using Flp recombinase-mediated site-specific recombination. An M3R-transfected CHO cell line, selected and propagated in hygromycin, was used to detect anti-M3R autoantibodies in SS patient and healthy control sera by flow cytometry. RESULTS: Testing of sera for the presence of anti M3R autoantibodies bound to CHO-transfected cells revealed the presence of anti M3R autoantibodies in SS patients (9 of 11) but not in healthy controls (0 of 11). Although the anti-M3R autoantibodies detected in patient sera were of multiple isotypes, the most consistently detected were IgG1, IgG3, and IgA. CONCLUSION: Using a newly constructed cell line expressing human M3R, anti-M3R autoantibodies were easily detected in sera from SS patients. These autoantibodies were skewed toward the IgG1, IgG3, and IgA isotypes, probably recognizing a tertiary epitope created by extracellular domains of the receptor protein. Anti-M3R autoantibodies represent a highly promising clinical marker for the identification of SS. PMID- 15334477 TI - What is the most appropriate radiologic scoring method for ankylosing spondylitis? A comparison of the available methods based on the Outcome Measures in Rheumatology Clinical Trials filter. AB - OBJECTIVE: To select the most appropriate radiologic scoring method for the evaluation of radiographic progression in ankylosing spondylitis (AS) in clinical trials. METHODS: The validity of the currently available methods, the Bath Ankylosing Spondylitis Radiology Index (BASRI), the Stoke Ankylosing Spondylitis Spine Score (SASSS), and the modified SASSS (M-SASSS), was tested according to the aspects of the Outcome Measures in Rheumatology Clinical Trials filter: truth, discrimination (reliability and sensitivity to change), and feasibility, using radiographs of 133 patients at 4 different time points (baseline, 1 year, 2 years, and 4 years). One observer scored these sets in chronological order. To assess interobserver reliability, a second observer scored radiographs of 20 patients at the 4 different time points. RESULTS: After 4 years, 9% and 8% of patients showed changes >0 in the sacroiliac (SI) joints and hips, respectively. Independent of the method chosen, approximately 40% of patients showed changes in both the lumbar and cervical spine. Therefore, it was concluded that, for the assessment of progression, SI joints and hips are of minor importance. The intraclass correlation coefficient (ICC) varied from 0.87 to 0.98 and ICCs for intraobserver scores varied from 0.96 to 0.99. Concerning progression scores, only the ICC for the M-SASSS measured after 2 years remained acceptable (0.82). The intraobserver scores for progression after 2 years of followup were an ICC of 0.93 for the BASRI, an ICC of 0.79 for the SASSS, and an ICC of 0.95 for the M SASSS. Concerning sensitivity to change, it was found that the M-SASSS classified the highest percentage of patients with a change >0. CONCLUSION: The M-SASSS is the most appropriate method by which to score the radiographic progression in AS patients in clinical trials. PMID- 15334478 TI - Central role of complement membrane attack complex in monosodium urate crystal induced neutrophilic rabbit knee synovitis. AB - OBJECTIVE: Monosodium urate monohydrate (MSU) crystals promote gouty inflammation that is critically mediated by neutrophil recruitment and activation. Interleukin 8 (IL-8) and closely related chemokines are major neutrophil chemotaxins in experimental gout. But MSU crystals also activate the classical and alternative pathways of complement, and MSU crystals directly cleave C5 on the crystal surface. Unlike IL-8, the roles in acute gout of individual complement-derived peptides and of the terminal C5b-9 complement components that comprise the membrane attack complex (MAC) are unclear. Hence, we studied rabbits deficient in the MAC component C6 to determine if MAC mediated urate crystal-induced arthritis. METHODS: We injected C6-deficient and C6-sufficient rabbit knee joints with 10 mg of pyrogen-free urate crystals and analyzed IL-8 levels, leukocyte influx, and joint inflammation 24 hours later. RESULTS: There was a significant decrease (>60%) in swelling in MSU crystal-injected knees of C6-deficient animals as compared with C6-sufficient animals (P < 0.05). An attenuated rise in MSU crystal-induced joint effusion levels of IL-8 also was observed, which was concordant with diminished numbers of neutrophils (P < 0.05) but not monocytes in MSU crystal-induced knee synovial fluid from C6-deficient animals. Synovial tissue analysis confirmed mononuclear leukocyte infiltration in response to MSU crystal injection in all animals, but substantial neutrophil infiltration only in C6-sufficient animals. CONCLUSION: MAC activation appears to play a major role in intraarticular IL-8 generation and in neutrophil recruitment in experimental acute gouty arthritis of the rabbit knee. C6 and MAC activation may represent novel therapeutic targets for suppression of neutrophil-mediated joint inflammation in gout. PMID- 15334479 TI - The genetic epidemiology of joint hypermobility: a population study of female twins. AB - OBJECTIVE: To estimate the genetic influence on joint hypermobility in an unselected population using a classic twin study design. METHODS: A self-report questionnaire on joint hypermobility as well as data on age, height, weight, estrogen replacement therapy, and menopause status were obtained from 483 monozygotic (MZ) and 472 dizygotic (DZ) unselected female twin pairs ages 21-81 years who were registered with the St Thomas' Adult Twin Registry in the UK. RESULTS: The overall prevalence of hypermobility was 19.5% in MZ twins and 22.1% in DZ twins. The prevalence of hypermobile joints declined with age, falling from 34% in subjects ages 20-30 years to 18.4% in those ages 60 years or older. Significantly greater concordance for joint hypermobility was observed in the MZ twins when compared with the DZ twins (60% versus 36%), consistent with a genetic influence. In variance components analysis, the age- and body mass index-adjusted heritability of joint hypermobility was estimated to be 70% (95% confidence interval 57-89%). CONCLUSION: Genetic factors have a substantial contribution to joint hypermobility in the adult female population. PMID- 15334480 TI - An analysis of CTLA-4 and proinflammatory cytokine genes in Wegener's granulomatosis. AB - OBJECTIVE: The precipitating event(s) that triggers Wegener's granulomatosis (WG) is unknown. Cytokines, costimulatory molecules, and counterregulatory molecules control the quality and intensity of immune responses. Thus, they are relevant candidates for genetic studies of immune dysregulation in WG, the pathogenesis of which may be facilitated by multiple acquired and/or inherited factors. This study was undertaken to investigate possible genetic associations of various proinflammatory cytokines and CTLA-4, a receptor for T cell inhibition, with WG. METHODS: Using polymerase chain reaction-based DNA genotyping, we investigated the polymorphisms located in the genes encoding a variety of proinflammatory cytokines and CTLA-4 in 117 American patients with WG and 123 ethnically matched healthy controls. RESULTS: Compared with controls, patients with WG had a significantly lower frequency of homozygosity for the shortest allele (designated allele 86) of the Ctla4 microsatellite polymorphism (AT)n located in the 3' untranslated region (3'-UTR) of exon 3 (47.0% versus 69.9%; P = 0.0005). Significant differences between patients and controls in the allelic and genotypic frequencies of polymorphisms in the other cytokine and cytokine receptor genes studied (tumor necrosis factor alpha [TNFalpha], TNF receptor I [TNFRI], TNFRII, interleukin-1beta [IL-1beta], IL-6) were not found. CONCLUSION: The Ctla4 (AT)n 86 allele has been previously demonstrated to be crucial for maintenance of normal levels of CTLA-4 expression and balance between T cell activation and inhibition. Our results in American patients confirm findings from a Scandinavian cohort in which a positive association between WG and longer alleles of (AT)n in the Ctla4 3'-UTR was demonstrated. Diminished frequencies of the most effective allele for CTLA-4 expression may represent a WG-related susceptibility mutation that accounts, in part, for increased T cell activation and clonal expansion in WG. Blockade of T cell costimulation using CTLA-4Ig might be a useful therapeutic intervention, providing an alternative or complementary approach to conventional treatment with immunosuppressive agents. PMID- 15334481 TI - Shedding of mutant tumor necrosis factor receptor superfamily 1A associated with tumor necrosis factor receptor-associated periodic syndrome: differences between cell types. AB - OBJECTIVE: To investigate the effect of mutations in tumor necrosis factor receptor superfamily 1A (TNFRSF1A) on the ability of the receptors to be cleaved from the cell surface upon stimulation. The mutations we studied are associated with clinically distinct forms of TNF receptor-associated periodic syndrome (TRAPS). We also investigated different cell types within the same form of TRAPS. METHODS: The shedding of TNFRSF1A in response to stimulation with phorbol myristate acetate was assessed in leukocytes and dermal fibroblasts from patients with C33Y TRAPS, and in HEK 293 cell lines stably transfected with constructs containing wild-type TNFRSF1A and/or TNFRSF1A mutants identified in TRAPS patients. RESULTS: The shedding of TNFRSF1A differed between cell types within the same form of TRAPS. In particular, dermal fibroblasts, but not leukocytes, from C33Y TRAPS patients demonstrated reduced shedding of TNFRSF1A. Shedding of both wild-type and mutant TNFRSF1A from the transfected HEK 293 cells showed minor differences, but was in all cases induced to a substantial extent. CONCLUSION: Differences in TNFRSF1A shedding are not purely a function of the TNFRSF1A structure, but are also influenced by other features of genetic makeup and/or cellular differentiation. It is unlikely that a defect in TNFRSF1A shedding per se can fully explain the clinical features that are common to TRAPS patients with different TNFRSF1A mutations. PMID- 15334482 TI - Dialysis-associated systemic fibrosis (nephrogenic fibrosing dermopathy): study of inflammatory cells and transforming growth factor beta1 expression in affected skin. AB - OBJECTIVE: Nephrogenic fibrosing dermopathy (NFD) is a newly recognized cutaneous fibrotic disorder occurring in individuals with end-stage renal disease (ESRD). The aim of the present study was to describe the clinical and histopathologic features of 9 new cases and to characterize the inflammatory cells and expression of transforming growth factor beta1 (TGFbeta1) in affected skin. METHODS: Clinical and laboratory assessments, including serology and pulmonary function studies, were performed in 9 patients undergoing long-term dialysis (8 hemodialysis; 1 peritoneal dialysis) for ESRD of diverse etiologies. Skin, fascia, striated muscles, lungs, and heart were examined by histopathology. Inflammatory cells were characterized by immunophenotyping using specific monoclonal antibodies. TGFbeta1 expression was determined by in situ hybridization. RESULTS: All patients displayed cutaneous features resembling both systemic sclerosis and diffuse fasciitis, with severe loss of motion and flexion contractures in multiple joints. Six patients displayed woody induration of the muscles of the legs, thighs, and forearms. Five of the 6 patients with lung involvement had a reduced diffusion capacity for carbon monoxide on pulmonary function testing. Marked elevations of the erythrocyte sedimentation rate and/or C-reactive protein level were found in 6 patients. Antinuclear antibodies were present at low titers in 4 patients. Histopathologic studies indicated that in addition to the dermis, the fibrotic process affected the subcutaneous tissue, fascia, striated muscles, lungs, and myocardium. Large numbers of CD68+/factor XIIIa+ dendritic cells and increased expression of TGFbeta1 were found in affected skin and muscle. CONCLUSION: Our findings indicate that the fibrotic process of NFD affects not only the dermis, but also the subcutaneous tissues, fascia, and other organs, including striated muscles, heart, and lungs. We therefore believe this is a systemic fibrosing process, and we suggest that dialysis-associated systemic fibrosis would be a better term for the condition. PMID- 15334483 TI - The prototypic tissue pentraxin PTX3, in contrast to the short pentraxin serum amyloid P, inhibits phagocytosis of late apoptotic neutrophils by macrophages. AB - OBJECTIVE: Phagocytosis of apoptotic cells can be facilitated by complement components and short pentraxins, such as serum amyloid P (SAP). In contrast, the long pentraxin PTX3 was shown to inhibit phagocytosis of apoptotic Jurkat cells by dendritic cells and to bind late apoptotic polymorphonuclear leukocytes (PMNs). Recently, levels of the pentraxin PTX3 were shown to parallel disease activity in small-vessel vasculitis, which is often characterized by leukocytoclasia, a persistence of leukocyte remnants in the vessel wall. We undertook this study to test our hypothesis that PTX3 inhibits phagocytosis of late apoptotic PMNs by macrophages, thereby leading to their accumulation in the vessel wall. METHODS: Macrophages were allowed to phagocytose late apoptotic or secondary necrotic PMNs that were incubated with or without PTX3 for 30 minutes prior to phagocytosis. Phagocytosis was allowed to occur in the presence of 30% normal human serum with or without SAP and with or without depletion of complement. To discriminate between an inhibitory effect of PTX3 on binding and the internalization of apoptotic PMNs into macrophages, internalization was blocked by cytochalasin B. RESULTS: SAP and complement were both necessary for effective in vitro phagocytosis. In contrast, PTX3 inhibited phagocytosis in a dose-dependent manner, from 11% inhibition at 6.25 microg/ml to almost complete inhibition at 100 microg/ml. Furthermore, PTX3 partly affected binding of apoptotic PMNs to macrophages. CONCLUSION: PTX3, in contrast to SAP and complement, inhibits phagocytosis of late apoptotic PMNs by monocyte-derived macrophages in a dose-dependent manner. Therefore, PTX3 can play a role in the development of leukocytoclasia by affecting the clearance of apoptotic PMNs, thereby inducing their accumulation in the vessel wall. PMID- 15334485 TI - Lack of requirement of osteopontin for inflammation, bone erosion, and cartilage damage in the K/BxN model of autoantibody-mediated arthritis. AB - OBJECTIVE: Osteopontin (OPN) is a secreted glycoprotein involved in a range of physiologic processes, including inflammation, immunity mediated by Th1 cells, and bone remodeling. It is expressed in the joints of rheumatoid arthritis patients and has been the subject of conflicting reports concerning its role in arthritis induced by antibodies against type II collagen. This study assessed the role of OPN in the K/BxN serum-transfer model of autoantibody-induced arthritis. METHODS: Expression of OPN gene transcripts was assessed by microarray analysis of ankle RNA taken at 6 time points after transfer of K/BxN serum. OPN-sufficient or OPN-deficient littermates backcrossed for 10 generations onto the C57BL/6 genetic background were given K/BxN serum. Arthritis severity was measured by ankle thickening and a clinical index. Hind limb sections were stained with hematoxylin and eosin or toluidine blue and scored for inflammation, cartilage damage, and bone erosion. RESULTS: OPN messenger RNA transcripts progressively increased in ankle joints during the course of K/BxN serum-transferred arthritis. OPN-deficient mice receiving K/BxN serum developed arthritis with kinetics and clinical severity comparable with those of OPN-sufficient littermates. Histologic assessment of arthritic joints from OPN-deficient mice revealed synovial hyperplasia, pannus formation, mononuclear cell infiltration, bone erosion, cartilage damage at sites adjacent to and distal from pannus invasion, and tartrate-resistant acid phosphatase-positive multinucleated cells at sites of bone erosion. Histopathologic scoring demonstrated comparable levels of inflammation, cartilage damage, and bone erosion in OPN-sufficient and OPN deficient mice. CONCLUSION: OPN does not have a required role in inflammation, bone erosion, and cartilage damage in the K/BxN serum-transfer model. PMID- 15334484 TI - Reduced infiltration and increased apoptosis of leukocytes at sites of inflammation by systemic administration of a membrane-permeable IkappaBalpha repressor. AB - OBJECTIVE: NF-kappaB activation is associated with several inflammatory disorders, including rheumatoid arthritis (RA), making this family of transcription factors a good target for the development of antiinflammatory treatments. Although inhibitors of the NF-kappaB pathway are currently available, their specificity has not been adequately determined. IkappaBalpha is a physiologic inhibitor of NF-kappaB and a potent repressor experimentally when expressed in a nondegradable form. We describe here a novel means for specifically regulating NF-kappaB activity in vivo by administering a chimeric molecule comprising the super-repressor IkappaBalpha (srIkappaBalpha) fused to the membrane-transducing domain of the human immunodeficiency virus Tat protein (Tat-srIkappaBalpha). METHODS: The Wistar rat carrageenan-induced pleurisy model was used to assess the effects of in vivo administration of Tat-srIkappaBalpha on leukocyte infiltration and on cytokine and chemokine production. RESULTS: Systemic administration of Tat-srIkappaBalpha diminished infiltration of leukocytes into the site of inflammation. Analysis of the recruited inflammatory cells confirmed uptake of the inhibitor and reduction of the NF-kappaB activity. These cells exhibited elevated caspase activity, suggesting that NF-kappaB is required for the survival of leukocytes at sites of inflammation. Analysis of exudates, while showing decreases in the production of the proinflammatory cytokines tumor necrosis factor alpha and interleukin-1beta, also revealed a significant increase in the production of the neutrophil chemoattractants cytokine-induced neutrophil chemoattractant 1 (CINC-1) and CINC-3 compared with controls. This result could reveal a previously unknown feedback mechanism in which infiltrating leukocytes may down-regulate local production of these chemokines. CONCLUSION: These results provide new insights into the etiology of inflammation and establish a strategy for developing novel therapeutics by regulating the signaling activity of pathways known to function in RA. PMID- 15334486 TI - Identification of two novel female-specific non-major histocompatibility complex loci regulating collagen-induced arthritis severity and chronicity, and evidence of epistasis. AB - OBJECTIVE: To identify additional sex-specific and epistatic quantitative trait loci (QTL) regulating collagen-induced arthritis (CIA) severity overall, as well as within different stages during the disease course, in an intercross between major histocompatibility complex-identical inbred rat strains DA/Bkl (susceptible) and ACI/Hsd (resistant). METHODS: Arthritic male (DA x ACI)F2 intercross offspring (n = 143) were analyzed separately from the females (n = 184). Phenotypic extremes (maximum arthritis scores [MAS]) were genotyped and used for QTL analysis. All 327 rats were genotyped with the simple sequence length polymorphism (SSLP) markers closest to the peak of Cia7 and Cia10, the major loci previously identified in this intercross, and with SSLPs covering chromosomes 12 and 18. Phenotypes studied were disease onset, arthritis severity scores on days 14-39, MAS, mean and cumulative arthritis scores, delayed-type hypersensitivity, and antibody responses to rat type II collagen. RESULTS: A new female-specific arthritis-severity recessive locus was identified on rat chromosome 12 (Cia25), with a maximum effect observed on day 28 (logarithm of odds [LOD] 4.7). The homozygous DA genotype at Cia25 was associated with a 45% higher median arthritis score in females. Sequencing analyses of the Cia25 candidate gene Ncf1 revealed polymorphisms between DA and ACI. The previously identified locus, Cia10, was found to be male-specific. A 2-locus interaction model analysis identified a novel recessive chromosome 18 QTL, Cia26, which was dependent on Cia7, with its maximum effect observed at later stages during the disease course (peak LOD score of 3.6 for arthritis scores on day 39). CONCLUSION: This study identified 2 novel female-specific loci, and 1 male specific locus. Cia25 regulates MAS and disease severity during the mid-to-late stages of the disease course and may be accounted for by Ncf1 polymorphisms. Cia26 is in epistasis with Cia7 and regulates later stages of disease, suggesting an involvement in disease perpetuation and/or chronicity. PMID- 15334487 TI - Lack of association between the corticotropin-releasing hormone locus and rheumatoid arthritis. PMID- 15334488 TI - Response to anakinra in a de novo case of neonatal-onset multisystem inflammatory disease. PMID- 15334490 TI - Monitoring guidelines for methotrexate-treated rheumatoid arthritis patients: comment on the article by Yazici et al. PMID- 15334491 TI - Intravenous immunoglobulin and antiphospholipid syndrome: comment on the article by Erkan et al. PMID- 15334493 TI - Dosage effects of orally administered bovine type I collagen on immune function in patients with systemic sclerosis. PMID- 15334494 TI - HLA-DQA1 as a risk factor for microchimerism: comment on the article by Artlett et al. PMID- 15334495 TI - Recommendations regarding individuals in whom bone densitometry should be performed: comment on the article by van Staa et al. PMID- 15334497 TI - No justification for publication of study on subgrouping of fibromyalgia patients: comment on the article by Giesecke et al. PMID- 15334498 TI - Tumor necrosis factor alpha blockade as therapy for sarcoidosis: comment on the article by Ulbricht et al. PMID- 15334500 TI - Variant chronic infantile neurologic, cutaneous, articular syndrome due to a mutation within the leucine-rich repeat domain of CIAS1. PMID- 15334502 TI - BEATing cystic fibrosis: more on rhDNase and mild lung disease. PMID- 15334503 TI - Longitudinal pulmonary status of cystic fibrosis children with meconium ileus. AB - Although meconium ileus (MI) is the earliest manifestation of cystic fibrosis (CF), and is associated with poorer growth, the longitudinal pulmonary progression of CF children with MI is not clear. To test the hypothesis that MI is associated with worse pulmonary outcomes, we prospectively compared from diagnosis to 12 years of age 32 CF children with MI to 50 CF children without MI who were diagnosed during early infancy through neonatal screening. Pulmonary outcome measures included respiratory symptoms, respiratory infections, pathogens, antibiotic usage, hospitalizations, quantitative chest radiology, spirometry, and lung volume determinations. Obstructive lung disease was defined as percent predicted spirometry values below the lower limits of normal. Longitudinal analyses revealed no significant differences in cough, wheezing, respiratory infections, prevalence of and median times to acquisition of Pseudomonas aeruginosa or Staphylococcus aureus, antibiotic usage, and chest radiograph scores between the two groups. However, MI children showed significantly worse forced expiratory volume in 1 sec (FEV(1)), forced vital capacity (FVC), forced expiratory flow between 25-75% of FVC (FEF(25-75)), % predicted FEV(1), % predicted FEF(25-75), and total lung capacity (TLC). These differences were particularly apparent beginning at age 8-10 years. MI children also had higher rates of and shorter median times to obstructive lung disease. Subgroup analyses showed MI children treated surgically and those treated medically had similar pulmonary outcomes. In conclusion, MI children have worse lung function and more obstructive lung disease than those without MI. Such abnormalities are accompanied by reduced lung volume. MI is a distinct CF phenotype with more severe pulmonary dysfunction. PMID- 15334504 TI - Treatment of experimental meconium aspiration syndrome with surfactant lung lavage and conventional vs. asymmetric high-frequency jet ventilation. AB - Respiratory failure caused by meconium aspiration requires combined strategies. We hypothesized that surfactant lung lavage with asymmetric high-frequency jet ventilation (AHFJV) can increase the removal of meconium and improve lung function. During conventional ventilation (CV), a suspension of human meconium (25 mg/ml, 4 ml/kg) was instilled into the tracheal tube of anesthetized rabbits to cause respiratory failure. Animals were then divided into four groups: saline lavage + CV (Sal-CV), surfactant lavage + CV (Surf-CV), saline lavage + HFJV (Sal HFJV), and surfactant lavage + HFJV (Surf-HFJV). Lung lavage (10 ml/kg in 3 portions) was performed with diluted surfactant (Curosurf, 100 mg of phospholipids/kg) or saline during CV (frequency (f), 30/min; inspiration time (Ti), 50%) or AHFJV (f, 300/min; Ti, 70%). Animals were ventilated for an additional hour with either CV or HFJV (Ti, 50%). Surfactant lavage with both CV and AHFJV removed more meconium than saline lavage. However, the highest removal was found in the Surf-HFJV group vs. all other groups (P < 0.05). The oxygenation index decreased after surfactant lavage in both groups compared to controls (P < 0.001), and more prominently in the Surf-CV group. Elimination of CO(2) was significantly higher in the Surf-HFJV group vs. all other groups (P < 0.05). The ventilation efficiency index increased after lavage in both surfactant groups vs. saline controls (P < 0.05). Dynamic lung-thorax compliance gradually increased, and right-to-left pulmonary shunts decreased in both surfactant groups vs. saline controls after lavage (P < 0.05). Combination of surfactant lavage with both CV and AHFJV was beneficial in rabbits with meconium aspiration syndrome. While AHFJV was more effective in the removal of meconium, CV had a more favorable effect on lung function in the postlavage period. PMID- 15334505 TI - Familial concordance of phenotype and microbial variation among siblings with CF. AB - The clinical spectrum of cystic fibrosis (CF) is influenced by the cystic fibrosis transmembrane conductance regulator (CFTR) genotype. However, variable courses of the disease were demonstrated among patients with identical genotypes. Since siblings share identical CFTR mutations and environmental factors, they can serve as a model to assess the effect of modifier genes on disease expression, and also to evaluate cross-infection. The aim of this study was to compare disease expression among siblings with CF. All sibling pairs treated at 7 CF centers in Israel were included in the study. Data were collected from patients' medical charts. Fifty families with at least 2 siblings were identified. As expected, the second-born sibling was diagnosed at an earlier age compared to the first-born. The mode of CF presentation at diagnosis showed significant familial concordance. In the families where the first sibling presented with gastrointestinal manifestations, 79% of the second siblings also presented with gastrointestinal manifestations. When gastrointestinal manifestations were absent in the first sibling, only 12% of the second siblings presented with gastrointestinal manifestations (P < 0.0001). Likewise, when the first sibling presented with respiratory symptoms, 60% of the second siblings presented with the similar symptoms. However, when the first sibling presented without respiratory symptoms, only 12% of the second siblings presented with respiratory symptoms (P < 0.001). Meconium ileus (MI) was present in 20 patients (21%). In 10 families where the first-born sibling had MI, 8 (80%) of the subsequent siblings had MI. On the other hand, in the 39 families where the first-born sibling did not have MI, only 2 (5%) subsequent siblings had MI (P < 0.001). Pancreatic insufficiency (PI) also had high familial concordance (P < 0.0001). Percentile growth for weights and heights and lung function (FVC, FEV(1), and FEF(25-75)) at ages 7 and 10 years were similar between siblings. P. aeruginosa grew from sputum in 89% of our study patients. When P. aeruginosa was isolated from the first-born patient, 91% of the second siblings were also positive for P. aeruginosa, whereas when the initial sibling was not a carrier of P. aeruginosa, only 50% of subsequent siblings were positive (P < 0.0001). This familial concordance was not observed for S. aureus. By contrast, the age of first isolation of P. aeruginosa and S. aureus was significantly earlier in the second sibling than in the first for the two bacteria: 10.3 +/- 5.1 vs. 7.3 +/- 5.2 years (P < 0.05) for P. aeruginosa, and 11.5 +/- 5.4 years vs. 6.8 +/- 5.1 years (P < 0.05) for S. aureus. CF siblings tend to share similar phenotypes that are not mutation dependent. The lack of variability between siblings in mode of initial CF presentation, rates of MI, pulmonary function, and nutritional status supports the role of modifier genes in the determination of these factors. PMID- 15334506 TI - Pediatric bronchiectasis: correlation of HRCT, ventilation and perfusion scintigraphy, and pulmonary function testing. AB - Bronchiectasis in children, although occurring with diminished frequency, continues as a major challenge for the pediatric pulmonologist. The method of choice for the diagnosis of the condition is high-resolution computed tomography (HRCT). The aim of the present study was to correlate the relationship(s) of HRCT, lung function, ventilation lung scintigraphy (VLS), and perfusion lung scintigraphy (PLS) in children with bronchiectasis. Sixteen children ranging in age from 4-18 years with clinical and chest X-ray evidence of bronchiectasis were enrolled in the study. The degree of bronchiectasis was assessed by HRCT scores, decrease in attenuation on expiratory scans, VLS, and PLS. HRCT scores for bronchiectasis and decreased lung attenuation showed a strong correlation with PLS (rho = 0.82; P < 0.001) and with VLS (rho = 0.72; P < 0.01). There was a moderate negative correlation between FEV(1) and HRCT bronchiectasis scores (rho = -0.53; P = 0.02), decreased lung attenuation score (rho = -0.64; P = 0.007), and atelectasis score (rho = -0.54; P = 0.03). In conclusion, HRCT provides a complete and precise assessment of children with bronchiectasis. Ventilation/perfusion scans and lung functions are additive tools to understand the complexity of the disease process and to improve diagnosis and therapeutic strategies. PMID- 15334507 TI - Measurement of inspiratory flow in children with acute asthma. AB - Dry-powder inhalers (DPIs) have been proposed for treatment of acute asthma. Different DPIs vary in their inspiratory resistance and have different recommended optimal peak inspiratory flows (PIFs). Reduced PIF during acute asthma may result in inadequate drug delivery to the lungs. Our aim was to measure the inspiratory flow in relation to inspiratory resistance during acute asthma in children presenting to the emergency room. School-age (range, 6-18 years) children were referred to the emergency room for acute asthma. PIF measurements were performed by In-Check Dial trade mark device with simulated airflow resistances equivalent to Turbuhaler, Diskus, and free flow. Percent change in PIF between remission and acute asthma (%Delta) was correlated with percent change in clinical score (CS) and percent change in spirometry in children <9 and >9 years old. Thirty-three children (21 males) participated. PIF with simulated Turbuhaler resistance was significantly lower than with simulated Diskus resistance in both acute and remission states (P < 0.0001). PIF with simulated Turbuhaler resistance increased from 62.1 +/- 15.3 (acute) to 74.4 +/- 16.5 l/min (remission, P < 0.0001), while with Diskus it rose from 72.6 +/- 20.5 to 91.1 +/- 18.9 l/min (P < 0.0001). Turbuhaler %Delta PIF correlated with %Delta FEV(1) (P = 0.01) and with %Delta CS (P = 0.0001). A lesser degree of correlation was observed while using Diskus resistance and in children above 9 years old. During acute asthmatic attacks, PIF is reduced; this reduction is particularly prominent in young children who use a high-resistance device. However, the PIF generated is generally within the values considered compatible with adequate lung deposition with both Diskus and Turbuhaler. PMID- 15334508 TI - Pulmonary lymphangiectasia: diagnosis and clinical course. AB - Pulmonary lymphangiectasia (PL) is a rare disease characterized by dilation of the normal lung lymphatics. The disease was previously associated with a poor prognosis. However, several recent case reports and case studies indicate that survival is possible in the severe neonatal form of PL, and the prognosis is good for infants who present after the neonatal period. In this report, we describe the etiology, clinical presentation, diagnosis, management, and outcome of PL. A revised classification of PL is proposed, based on the current state of knowledge of its etiology and presentation. PMID- 15334509 TI - Treatment with tobramycin solution for inhalation reduces hospitalizations in young CF subjects with mild lung disease. AB - Our objective was to study the effect of tobramycin solution for inhalation (TSI; TOBI, Chiron Corp.) on lung function decline rate in 400 young persons with cystic fibrosis (CF) and mild lung disease. Effects on hospitalization, antibiotic use, school days missed, and nutritional status also were determined. This was an open-label, randomized (stratified by sex and age group, i.e., 6-10 and 11-15 years), parallel-group, multicenter study. Routine subject management (control group) was compared to routine management plus 28 days of twice-daily TSI inhalation, followed by 28 days off the drug (TSI group) for 56 weeks. Primary efficacy endpoints included rate of lung function decline (as measured by forced expiratory volume in 1 sec; FEV(1)), hospitalization, and concomitant antibiotic use. Safety was assessed by analysis of treatment-emergent adverse events. Only 184 of 400 planned subjects were recruited and randomized (93 to the TSI group, and 91 to the control group). Enrollment was ended after 2 years because of difficult recruitment. An interim safety review showed a 2.42-fold risk of respiratory hospitalization for control group subjects (P = 0.020), and the study was terminated. Sixty-three subjects (34.2%) completed the entire study (30 in the TSI group, or 32.3%; and 33 in the control group, or 36.3%). Significantly fewer TSI subjects were hospitalized for worsening of respiratory symptoms (11.0% vs. 25.6%; P = 0.011), and fewer TSI subjects were hospitalized overall (16.5% vs. 27.8%; P = 0.065). Fewer TSI subjects received antibiotics other than the study drug (78.0% vs. 95.6%), and significantly fewer received oral antibiotics (76.9% vs. 91.1%; P = 0.009). No other safety or adverse event differences were observed. In conclusion, significant reductions in respiratory hospitalizations, concomitant antibiotic use, and a trend towards improvement in percent predicted forced expiratory flow (FEF(25-75)) provide evidence of a clinical benefit of TSI use in young persons with CF and mild lung disease. An effect on lung function decline rate could not be evaluated as planned, due to inadequate enrollment and early study termination. PMID- 15334510 TI - Familial susceptibility to severe respiratory infection in early life. AB - Lower respiratory tract infections (LRTI) are common in the first year of life and are mostly caused by viruses. Severity of LRTI in infants is associated with early-life environmental factors. Genetic association studies also suggest a role of heredity in susceptibility to acute bronchiolitis. We designed a case control study to further investigate relative importance of familial influences in risk of LRTI in early childhood compared to environmental factors. From a hospital database, we selected 1,308 children (436 cases; 872 controls) living in Oxfordshire. Cases were children under age 5 years admitted to hospital with LRTI. Parental history and other exposures were recorded in cases and controls by postal questionnaire. Maternal history of asthma increased the risk of severe LRTI in the first year of life, independent of subsequent asthma in a child. History of maternal bronchiolitis also increased the risk of infant LRTI. These results further support the possibility that genetic factors play an important role in susceptibility to severe viral respiratory infections in early life, and suggest that this effect may be independent of subsequent childhood asthma. PMID- 15334511 TI - Infant immunization and the occurrence of atopic disease in Dutch and German children: a nested case-control study. AB - Our goal was to assess the role of early childhood vaccination in the occurrence of respiratory symptoms and allergic sensitization in 7-8-year-old Dutch and German children. A nested case-control study was conducted among children participating in a large longitudinal study on respiratory health, to study the relationship between vaccination (bacille Calmette-Guerin (BCG), pertussis, measles/mumps, rubella, and Haemophilus influenza type b (Hib)) and respiratory symptoms and allergic sensitization. Parents of 510 7-8-year-old children with respiratory complaints and an equal number of randomly selected children without respiratory complaints were asked to complete a questionnaire. Blood samples were collected for specific serum IgE analysis. Vaccination status was assessed through the records of the participating Municipal Health Services. No association between vaccination against pertussis, measles, rubella, or Hib and respiratory symptoms or allergic sensitization was found. For sensitization against house dust mite, BCG vaccination resulted in an increased risk (OR, 2.28; 95% CI, 1.05-4.96). Birth order was inversely associated with allergic sensitization, but was not related to respiratory symptoms. We found an association between BCG vaccination and the subsequent risk for sensitization against house dust mite. No evidence was found for an association between vaccination and respiratory symptoms. Earlier reports of an association of birth order with atopic disease were supported by the results of the present study. PMID- 15334512 TI - Effect of inhalation times on exhaled NO. AB - Recent work suggests that exhaled nitric oxide (eNO) can be divided into airway and alveolar components, using varying expiratory flow rates. It is known that higher values of eNO are found with lower expiratory flow rates, thought to be due to prolonged contact of the expirate with the bronchial epithelium. However, whether a prolonged inspiratory time could contaminate the alveolar signal with bronchial derived NO, by the same mechanism, is unknown. We aimed to study the effect, including repeatability, of inhalation time on eNO measurements performed at three different flow rates. We measured eNO in 21 children with asthma of different severity (no steroids, n = 2; inhaled corticosteroid (IC) dose up to 200 mcg/day, n = 3; IC up to 500 mcg/day, n = 3; IC >500 mcg/day, n = 6; IC >500 mcg/day and oral steroid, n = 7) and 24 normal adult controls at 50, 100, and 200 ml/s expiratory flow rate. The effect of either a rapid or a slow inhalation on measured eNO was studied at each flow rate. Furthermore, 12 asthmatic children and 12 adults had repeated measurements 1 hr apart. Repeatability within 1 day was assessed by calculating the single-determination standard deviation (SD) and 95% range. Our results showed that repeatability was equally good for the three expiratory flow rates, and inhalation time had no influence on the results. The inhalation maneuver does not influence eNO measurements using the variable expiratory flow technique. PMID- 15334513 TI - Lung growth and function in children and adolescents with idiopathic pectus excavatum. AB - Patterns of lung growth and function were studied retrospectively in 103 patients (73 male and 30 female) with idiopathic pectus excavatum in order to determine: 1) the prevalence of restrictive lung defect and/or other lung function abnormalities; 2) the possible association between type of lung function abnormalities and age of the patient; and 3) whether the type of lung function is associated with other clinical conditions. Forty-three patients (42%) were between 5-9 years of age; 36 (35%) were between 10-14 years; and 24 (23%) were between 15-19 years. Restrictive lung disease was detected only in 5 patients (5%), whereas 42 patients (41%) had evidence of obstructive pattern, and the remaining 56 patients (54%) had a normal pattern. The mean values for lung volume (total lung capacity (TLC) and/or forced vital capacity (FVC)) were lowest in the 10-14-year group, whereas the mean values of the indices of lower airway function (forced expiratory volume at 1 sec (FEV(1)), FEV(1)/FVC, forced expiratory flow at 25-75% of forced vital capacity (FEF(25-75)), and FEF(25-75)/FVC) were higher in the younger group. Residual volume (RV) and RV/TLC were elevated in all age groups, but they declined toward normalization with increasing age. There were no significant differences between groups (stratified either by pattern of lung function or by age) with regard to their demographics or clinical characteristics (scoliosis, direction of sternal rotation, history of asthma/reactive airways disease, or exercise intolerance). We conclude that idiopathic pectus excavatum may be associated with a variety of lung function abnormalities (in particular, lower airway obstruction) even in the absence of overt clinical symptoms. There was no evidence of significant worsening of lung function with increasing age. PMID- 15334514 TI - Celiac disease and pulmonary hemosiderosis in a patient with chronic granulomatous disease. AB - We report on a patient with the hitherto undescribed combination of chronic granulomatous disease, pulmonary hemosiderosis, and celiac disease. The hemosiderosis resolved with a gluten-free diet and glucocorticosteroid pulse therapy, but the restrictive lung function pattern remained unchanged. Lung function improved markedly by immunosuppression with daily glucocorticosteroid and azathioprine treatment. PMID- 15334515 TI - PHOX2B gene mutation in a patient with late-onset central hypoventilation. AB - Congenital central hypoventilation syndrome, which is related to abnormal autonomic control of breathing and typically manifests at birth, was recently associated with PHOX2B gene mutations. In contrast, central hypoventilation with later onset constitutes a poorly defined group of unknown etiology. Here, we report on the identification of a de novo heterozygous PHOX2B mutation in a patient with central hypoventilation manifesting in childhood. This finding suggests that some of these cases may be genetically determined and allelic to congenital central hypoventilation syndrome. PMID- 15334516 TI - Recurrent respiratory papillomatosis in a survivor of extreme prematurity. AB - Recurrent respiratory papillomatosis (RRP) is a rare disease in children. Previous reports suggested that prematurity and early age of presentation were poor prognostic factors. We report on a 24-week premature infant who presented with stridor, weak cry, and hoarseness of voice at age 9 months (corrected age), in whom the diagnosis of RRP was not made until age 21 months (corrected age). Laser excision of RRP was subsequently performed, and the child is still surviving at age 2.5 years. RRP should be considered in the differential diagnosis of airway problems in survivors of extreme prematurity; the prognosis is not uniformly poor in premature infants. PMID- 15334517 TI - Acute respiratory distress due to fibrosarcoma of the carina in a child. AB - We report a 12-year-old boy with primary bronchopulmonary fibrosarcoma (PBPF). He was misdiagnosed a having asthma until he presented with acute respiratory distress syndrome. Chest x-ray showed atelectasis of the right lung. Bronchoscopy performed to rule out foreign body inhalation revealed a friable mass obstructing the right main bronchus. Successful treatment was achieved with surgical resection of the lesion followed by radiotherapy and chemotherapy. PMID- 15334518 TI - Re: Repeatability of sodium and chloride in exhaled breath condensates, Zacharasiewicz et al. Pediatr Pulmonol 2004;37:273-275. PMID- 15334519 TI - To the editor: Re: "Mucoepidermoid carcinoma of the bronchus presenting with a negative chest X-ray and normal pulmonary function in two teenagers: two case reports and review of the literature" by Robert J. Giusti and Raja M. Flores (Pediatric Pulmonology 2004;37:81-84). PMID- 15334522 TI - Effect of long-acting beta2 agonists on exacerbation rates of asthma in children. PMID- 15334524 TI - NTP-CERHR Expert Panel Report on the reproductive and developmental toxicity of fluoxetine. PMID- 15334525 TI - Juvenile animal studies: testing strategies and design. PMID- 15334526 TI - Dose-response for retinoic acid-induced forelimb malformations and cleft palate: a comparison of computerized image analysis and visual inspection. AB - BACKGROUND: The objectives of this study were to (1) compare two techniques (computerized image analysis and visual morphological evaluation) for the assessment of fetal forelimb malformations and (2) increase the robustness of the dose-response curve for forelimb and cleft palate malformations resulting from all-trans retinoic acid (RA) exposure in GD 11 mice. METHODS: Pregnant CD-1 mice were administered a single oral dose of all-trans RA (0, 2.5, 10, 30, 60, or 100 mg/kg) on GD 11. GD 18 fetuses were examined for malformations using visual morphological scoring and computerized image analysis. RESULTS: Dose-dependent changes occurred in the size and shape of the humerus, radius, and ulna based on both assessment methodologies. The most sensitive indicators for the lowest effect level (10 mg/kg) on forelimbs were roundness, a shape measurement determined by image analysis, and visual morphological scoring. For all other bone measurements (proximal and distal width, area, length, and perimeter), the lowest effect level was 30 mg/kg. The maximum effect for limb defects and total malformed fetuses was seen at 60 mg/kg and higher. Incidence of cleft palate increased over the entire range of administered doses reaching a maximum of 74% (100 mg/kg). CONCLUSIONS: Overall, results indicate that computerized image analysis was no more sensitive in detecting changes in the humerus, radius, and ulna than gross visual examination. Dose-response modeling of developmental endpoints yielded comparable benchmark dose levels for long bones and cleft palate that ranged from 0.24 to 7.6 mg/kg all-trans RA. Birth Defects Res B 71:289-295, 2004. PMID- 15334527 TI - Developmental toxicity evaluation of butylparaben in Sprague-Dawley rats. AB - The developmental toxicity potential of butylparaben (CAS No. 94-26-8) was evaluated in rats. Sprague-Dawley rats were administered butylparaben in 0.5% carboxymethylcellulose by oral gavage at dose levels of 0, 10, 100, or 1,000 mg/kg/day on gestation days (GD) 6-19 (sperm positive day = GD 0). Caesarean sections were performed on GD 20 and fetuses were evaluated for viability, growth, and external, visceral, and skeletal abnormalities. Each group consisted of 25 females, with at least 21 per group being pregnant. The highest dose level caused decreases in maternal weight gain during some of the measurement intervals and was statistically significant during the GD 18-20 interval. Maternal food consumption was significantly decreased in the highest dose group over the dosing period (GD 6-20). There were no differences from control in any of the developmental parameters measured, including embryo/fetal viability, fetal weight, malformations, or variations. Based on the results of this study, the maternal NOAEL for butylparaben was 100 mg/kg/day. Butylparaben does not have the potential to cause developmental toxicity in the Sprague-Dawley rat at oral dosages up to 1000 mg/kg/day. PMID- 15334528 TI - Health-related quality of life in women previously treated for early-stage breast cancer. AB - This study describes health-related quality of life (HRQOL) in women previously treated for early stage breast cancer relative to general population and breast cancer norms. It also identifies correlates and multivariate predictors of physical and mental HRQOL. 2582 women who were up to four years post-treatment for early stage breast cancer completed the RAND-36 Health Survey and reported personal and cancer-related information. Participants reported generally high HRQOL that was comparable to norms for women in the general population and other women with breast cancer. In multivariate analyses, better physical HRQOL was associated with fewer psychological symptoms, lower body mass index, better sleep quality, and more physical activity. Better mental HRQOL was associated with better sleep quality, fewer life events, less pain, and fewer gastrointestinal symptoms. Interventions targeting psychosocial symptoms, weight reduction, physical activity, sleep hygiene, and pain could result in improved HRQOL in these women. PMID- 15334529 TI - Exploring comprehensibility and manageability in palliative home care: an interview study of dying cancer patients' informal carers. AB - The presence of an informal carer is often a prerequisite for successful palliative home care, and the staff's ability to support informal carers' coping in such situations is important. Recent research has revealed that it is possible to achieve positive psychological states in palliative care despite the burdening situation. As there is a lack of theory-based coping studies, the aim of this study was to describe, within the context of palliative home care, two concepts in Antonovsky's theory of Sense of Coherence: comprehensibility (a perception that the challenge is understood) and manageability (a perception that the resources to cope are available). Tape-recorded semi-structured interviews with 19 informal carers during ongoing palliative home care were transcribed and analysed with a qualitative hermeneutic approach. Elements that facilitated comprehensibility included open information, symbolic information, basic life assumptions and previous knowledge. These were important for creating a congruent inner reality (as opposed to chaos). Resources contributing to manageability dealt with power, support, competence and accessibility, which on a more abstract level resulted in a feeling of togetherness (as opposed to isolation). The findings are discussed in relation to the complexity of communication between staff and carers within palliative care. PMID- 15334530 TI - Multiple risk behaviors among smokers in the childhood cancer survivors study cohort. AB - The literature on health behaviors of young adult cancer survivors is very limited, and thus little is known about preventable risk factors in this population. This paper describes the prevalence of five behavioral risk factors among 541 young adult survivors of childhood cancers from the CCSS cohort who were identified as smokers and enrolled in a randomized controlled trial of a smoking cessation intervention. The relationship between presence of multiple risk factors and a number of smoking-related factors was examined. About 31% of the sample engaged in zero or one health-risk behavior in addition to smoking; 63% engaged in 2 or 3, and 6% engaged in 4 or 5. There were positive linear relationships between number of risk factors and smoking rate and nicotine dependence. Number of risk factors was not associated with self-efficacy for quitting, but was related to readiness to quit. This study demonstrated that childhood cancer survivors who smoke have a number of other risk factors for the development of preventable disease and the presence of these risks was associated with factors that decrease the likelihood of quitting smoking. Attention to other health behaviors may be an important strategy for helping smokers quit. In particular, helping childhood cancer survivors who smoke to reduce other risk behaviors might also encourage them to quit smoking. PMID- 15334531 TI - Predictors and mediators of psychological adjustment in mothers of children newly diagnosed with cancer. AB - Early psychological adjustment (PA) of mothers of children with cancer (MCC) and mothers of children with acute illnesses (MCA) were compared, and predictors and mediators of maternal adjustment were tested. Sixty-nine MCC and 22 MCA completed standardized measures of depression, anxiety, global mental health (GMH), concurrent stress and strains, social support, coping strategies and child behavior. MCC reported greater depressive symptomatology, emotion-focused coping, and social support than did MCA. For MCC, emotion-focused coping and child behavior both predicted depression, anxiety, and GMH; concurrent stress and strain mediated the relationship between child behavior and depression and between emotion-focused coping and each measure of adjustment. For MCA, only emotion-focused coping predicted PA and concurrent stress and strains mediated the relationship between emotion-focused coping and depression. These findings suggest that MCC have more PA difficulties that are uniquely related to their child's behavior than MCA. Concurrent stress and strains seems to be an important mediator of PA for both groups of mothers. PMID- 15334532 TI - Development and pilot testing of a psychoeducational intervention for oral cancer patients. AB - OBJECTIVE: Oral cancer elicits considerable distress in both the pre and post treatment periods. This paper details the development, validation and pilot testing of a psychoeducational intervention for oral cancer patients. METHOD: An educational booklet covering information about oral cancer and its treatment and effective coping strategies was developed by a multi-disciplinary team. It was evaluated by patient focus groups and content/educational experts, delivered to subjects in pre and post-operative presentations by a health educator, and pilot tested in a randomized controlled trial versus standard care. RESULTS: Pilot data from 19 subjects (10 psychoeducation, nine standard care) indicates that the intervention is feasible and highly acceptable. At follow-up the intervention group showed a gain in knowledge, less body image disturbance, lower anxiety and a trend toward higher wellbeing. CONCLUSION: This program, which is currently being evaluated in a larger RCT with extended follow-up, should prove useful in reducing the psychosocial burden of oral cancer and its treatment. PMID- 15334533 TI - The decision to have a prosthesis: A qualitative study of men with testicular cancer. AB - The aim of this paper is to understand why some men decide to have a testicular prosthesis while others reject the idea and to explore the extent to which the option of a testicular prosthesis is adequately presented. Forty-five men with testicular cancer were interviewed about all aspects of their illness. Some men decided against a prosthesis for a number of reasons; they thought that the loss of a testicle was not visibly obvious, did not affect self-image, was not a long term threat to masculinity, and that living with one testicle was comfortable. Some did not want additional surgery and others were concerned about safety issues. The men who decided to have a prosthesis were concerned about self-image and a threat to masculinity. They had a desire to look 'normal' and a wish to keep the orchidectomy secret. Some men felt that the decision had been rushed. Others wished that they had had information about the possibility of implants before the orchidectomy or they considered the information presented inadequate. We conclude that it is important to counsel men with testicular cancer about the advantages and disadvantages of having a testicular prosthesis and to give them time to consider issues such as cosmetic appearance, self-image, comfort, possible short term complications and long-term safety. PMID- 15334534 TI - Cancer-related attitudes: A comparative study in Japan and the US. AB - The cancer-related attitudes and beliefs of adults living in central Japan (N = 357) were compared to those of respondents in a US state (Hawaii) (N = 223) consisting of both Japanese (n = 106) and Caucasian (n = 117) individuals. Almost all US subjects endorsed doctors disclosing cancer diagnoses to their patients, while a minority of the Japan sample supported cancer disclosure as a general practice. However, the majority of Japan respondents expressed a personal preference to be told if they themselves were diagnosed with cancer, as did virtually all US respondents. US subjects were more optimistic than Japanese subjects about cancer's curability, both at the present time and in the future; Japanese Americans scored intermediate between Japanese and Caucasian Americans. Word-association data indicated that cancer was most likely to be associated with death by Japanese respondents. The data support the continued international differences in views about cancer and suggest that some of these differences persist over many generations of US residence. PMID- 15334535 TI - Smallpox: the old and the new. PMID- 15334536 TI - An attempt at a new analysis of the mortality caused by smallpox and of the advantages of inoculation to prevent it. 1766. PMID- 15334537 TI - Therapeutic potential of nucleoside/nucleotide analogues against poxvirus infections. AB - Several nucleoside and nucleotide analogues have been identified as potent antiviral agents with convincing activity against poxviruses (including variola, vaccinia, monkeypox, cowpox, molluscum contagiosum, orf). Among the nucleoside analogues, 8-methyladenosine and 2-amino-7-[(1,3-dihydroxy-2 propoxy)methyl]purine (S2242), have been identified as promising anti-poxvirus agents. Among the nucleotide analogues, (S)-1-(3-hydroxy-2 phosphonylmethoxypropyl)cytosine [(S)-HPMPC, cidofovir], (S)-9-(3-hydroxy-2 phosphonylmethoxypropyl)-2,6-diaminopurine [(S)-HPMPDAP] and (S)-6-(3-hydroxy-2 phosphonylmethoxypropyl)oxy-2,4-diaminopyrimidine [(S)-HPMPO-DAPy] have been identified as promising anti-poxvirus agents. These nucleoside and nucleotide analogues have proved to be efficacious in various animal models for poxvirus infections. Only one compound, cidofovir, has also proved efficacious against poxvirus infections in humans, i.e. molluscum contagiosum and orf (sheep pox). Cidofovir is formally licensed for clinical use (intravenous administration for the treatment of cytomegalovirus retinitis in AIDS patients); however, it could also be formulated for topical administration, or for oral administration in prodrug form, i.e. as 1-O-hexadecyloxypropyl-cidofovir. PMID- 15334538 TI - Human tumor suppressor p53 and DNA viruses. AB - Human tumor suppressor protein p53 plays a major role in the cell cycle, orchestrating a number of important genes involved in cell-cycle control and apoptosis, and seems to be one of the most important molecules protecting cells from malignant transformation. Mutations in the p53 gene are observed in about 50% of primary tumors, inducing defective p53 protein no longer capable of binding DNA and of activating transcription. Certain DNA viruses are thought to act in a similar way and may also contribute to the progression of invasive cancer in infected tissue. One of the most effective strategies employed by these viruses is the inhibition of p53 protein by interaction with viral oncoproteins, implying a direct but also an indirect role of these viruses in the impairment of p53 structure and function. This article provides a summary of current knowledge concerning p53 tumor suppressor protein and reviews the different mechanisms adopted by different DNA viruses in undermining p53 function. PMID- 15334539 TI - HIV fusion and its inhibition in antiretroviral therapy. AB - The end of the twentieth century saw dramatic improvements in the prognosis of HIV infection brought about by the introduction of new agents (the protease inhibitors and the non-nucleoside reverse transcriptase inhibitors) and their use in highly active combinations. However, the durability of these combination treatments is limited by a number of factors including adverse effects and extensive intra-class cross-resistance so that new antiretrovirals acting on alternative targets and having improved systemic tolerability profiles are required. The HIV binding and entry process offers several potential targets for antiviral interaction. These include gp120 binding to CD4 and to chemokine co receptor molecules as well as the fusion process itself, which involves interactions between two leucine zipper-like 4-3 repeat regions within gp41 known as heptad repeat (HR)1 and HR2. Peptides such as enfuvirtide (formerly DP178 or T 20), that mimic the HR2 region of gp41, inhibit HIV-1 by a mechanism that is thought to involve competitive binding to HR1. This review summarises the clinical development of enfuvirtide, providing an overview of the pharmacokinetic, efficacy and safety data in various patient populations, and also considers the evidence for the key role of genotypic changes in the HR1 region (amino acids 36-45) in determining viral susceptibility to inhibition by enfuvirtide. PMID- 15334541 TI - Involvement of fumarate hydratase in nonsyndromic uterine leiomyomas: genetic linkage analysis and FISH studies. AB - Recently, germline mutations of the fumarate hydratase (FH) gene, in 1q42.1, have been found to be involved in syndromes associated with uterine leiomyomas (ULs). Compelling evidence also supports a genetic liability to develop nonsyndromic UL, although susceptibility genes have not been reported to date. Loss of heterozygosity (LOH) studies have found no or rare evidence of LOH of FH in nonsyndromic UL. However, the karyotypes of these tumors were not reported, and cytogenetic aberrations of 1q42-44 have been observed infrequently in UL. To determine whether FH mutations also may predispose women to developing nonsyndromic UL, we performed a genetic linkage study with DNA from 123 families containing at least one affected sister pair. In addition, to assess the frequency of FH loss specifically in UL with 1q rearrangements, we performed a fluorescence in situ hybridization (FISH) analysis of UL with 1q rearrangements. Analysis of the genotyping data revealed evidence suggestive of linkage to the FH region among study participants who were less than 40 years of age at diagnosis (Zlr 1.7 at D1S547, P = 0.04). FISH results showed that one copy of FH was absent in 9 of 11 ULs. These data indicate that loss of FH might be a significant event in the pathogenesis of a subset of nonsyndromic ULs. PMID- 15334542 TI - Comparative expressed sequence hybridization studies of high-hyperdiploid childhood acute lymphoblastic leukemia. AB - The functional consequences of a high-hyperdiploid karyotype, found in up to one third of cases of acute lymphoblastic leukemia (ALL), are unknown. Using the technique of comparative expressed sequence hybridization (CESH), we sought to address the question of whether increased chromosome copies in hyperdiploid ALL lead to increased gene expression. Relative expression of hyperdiploid ALL blasts versus peripheral blood mononuclear cells was analyzed in 18 patients. Common regions of overexpression corresponding to the presence of tri-/tetrasomies included: Xp22.1-22.2, 4q28, 6q14-15, 6q24, 10p13, 14q23-24, 17q21, 18q12, and 21q21, identified in 28-89% of cases. However, increased expression without underlying trisomy occurred at 3p21.3, 7q11.2, 8p21, and 8q24.1 in 39-90% of cases. High expression at 7q11.2, the most consistent change detected, was confirmed by quantitative PCR. Poor correlation between the presence of tri /tetrasomy and overexpression was observed for chromosomes 14 and 17. Two cases were reanalyzed versus (i) B cells, (ii) transformed B cells, and (iii) CD34+19+ cells (the putative counterpart of the leukemic cell). A reduction in the number of relatively overexpressed regions was observed with CD34+19+ cells. In particular, the peak at 7q11.2 disappeared, suggesting up-regulation of genes from this region in the early ontology of normal B-cell development. In conclusion, we have shown that tri-/tetrasomies in hyperdiploid ALL lead to an increase in the expression of associated sequences. The choice of a biologically relevant reference is crucial for data interpretation. PMID- 15334543 TI - Structural integrity and expression of the L3MBTL gene in normal and malignant hematopoietic cells. AB - The human L3MBTL gene is located in 20q12, a region that is commonly deleted in myeloproliferative disorders (MPD), myelodysplastic syndromes (MDS), and acute myeloid leukemia (AML). L3MBTL is highly homologous to the D-lethal(3) malignant brain tumor [D-l(3)mbt] gene, which is a putative tumor-suppressor gene (TSG) identified in Drosophila and which is closely related to the Drosophila sex combs on midleg (SCM) protein, a member of the Polycomb group (PcG) family of transcriptional repressors. To examine whether L3MBTL functions as a "classic" TSG in human hematologic malignancies, we screened a panel of 17 myeloid leukemia cell lines and peripheral blood or bone marrow samples from 29 MDS and 13 MPD patients for mutations in the entire L3MBTL coding sequence, including intron/exon splice junctions. No mutations were identified, although two single nucleotide differences were found (in intron 14 and in exon 15), which were interpreted as polymorphic changes. We used real-time RT-PCR to quantify the level of L3MBTL mRNA in various normal myeloid and lymphoid cell populations. L3MBTL is expressed in normal CD34+ bone marrow cells, and we found that the pattern of L3MBTL expression was similar to that of BMI1, a well-studied PcG gene with oncogenic activity, suggesting that L3MBTL and BMI1 may be co-regulated during hematopoiesis. The expression of L3MBTL mRNA in 30 of 35 cell lines and 13 of 15 AML samples was comparable to the level of L3MBTL expression in the normal cell populations. However, five leukemia cell lines showed no L3MBTL expression, and two of the AML samples showed aberrant L3MBTL expression. These data suggest that L3MBTL is not mutated in MDS or MPD. However, given the known dosage effects of PcG proteins in regulating gene expression, reduced or absent L3MBTL expression may be relevant in some cases of myeloid leukemia. PMID- 15334544 TI - Aberrations of breast cancer susceptibility genes occur early in sporadic breast tumors and in acquisition of breast epithelial immortalization. AB - In the search for early deletion targets in sporadic breast cancer, the analysis of TP53, BRCA1, BRCA2, and ATM revealed loss of heterozygosity (LOH) in tumor cells at 1 or more genes in 18 of 24 cases examined. Notably, in more than 60% of such tumors, LOH was detectable in morphologically normal terminal ductal lobular units (TDLUs) adjacent to carcinoma (LOHint). At BRCA2 and ATM, LOHint was most frequent, particularly in TDLUs of women 2 in apoptotic cells, in accordance with observed cell shrinkage, and changed little in necrotic counterparts, where only slight swelling was evident. These results indicate that PFG 1H-NMR serves as a sensitive indicator of early cell death in monolayer cultures, and can be used to distinguish apoptosis from necrosis. Measurements of restricted diffusion and water exchange are presented to elucidate the compartment origins and justify the model assumptions. PMID- 15334568 TI - Intracellular manganese ions provide strong T1 relaxation in rat myocardium. AB - The efficacy of manganese ions (Mn2+) as intracellular (ic) contrast agents was assessed in rat myocardium. T1 and T2 and Mn content were measured in ventricular tissue excised from isolated perfused hearts in which a 5-min wash-in with 0, 30, 100, 300, or 1000 microM of Mn dipyridoxyl diphosphate (MnDPDP) was followed by a 15-min wash-out to remove extracellular (ec) Mn2+. An inversion recovery (IR) analysis at 20 MHz revealed two T1 components: an ic and short T1-1 (650-251 ms), and an ec and longer T1-2 (2712-1042 ms). Intensities were about 68% and 32%, respectively. Tissue Mn content correlated particularly well with ic R1-1. A two site water-exchange analysis of T1 data documented slow water exchange with ic and ec lifetimes of 11.3 s and 7.5 s, respectively, and no differences between apparent and intrinsic relaxation parameters. Ic relaxivity induced by Mn2+ ions in ic water was as high as 56 (s mM)(-1), about 8 times and 36 times higher than with Mn2+ aqua ions and MnDPDP, respectively, in vitro. This value is as high as any reported to date for any synthetic protein-bound metal chelate. The increased rotational correlation time (tauR) between proton and electron (Mn2+) spins, and maintained inner-sphere water access, might make ic Mn2+ ions and Mn2+ -ion releasing contrast media surprisingly effective for T1-weighted imaging. PMID- 15334569 TI - Quantification of human atherosclerotic plaques using spatially enhanced cluster analysis of multicontrast-weighted magnetic resonance images. AB - One of the current limitations of magnetic resonance imaging (MRI) is the lack of an objective method to classify plaque components. Here we present a cluster analysis technique that can objectively quantify and classify MR images of atherosclerotic plaques. We obtained three-dimensional (3D) images from 12 human coronary artery specimens on a 9.4T imaging system using multicontrast-weighted fast spin-echo (T1-, proton density-, and T2-weighted) imaging with an isotropic voxel size of 39 micro. Spatially enhanced cluster analysis (SECA) was performed on multicontrast MR images, and the resulting segmentation was evaluated against histological tracings. To visualize the overall structure of plaques, the MR images were rendered in 3D. The specimens exhibited lesions of American Heart Association (AHA) plaque classification types I-VI. Both MR images and histological sections were independently reviewed, categorized, and compared. Overall, the classification obtained from the cluster-analyzed MR and histopathology images showed very good agreement for all AHA types (92%, Cohen's kappa = 0.89, P < 0.0001). All plaque types were identified and quantified by SECA with a high degree of correlation between cluster-analyzed MR and manually traced histopathology data. MRI combined with SECA provides an objective method for atherosclerotic plaque component characterization and quantification. PMID- 15334570 TI - Detection and volume determination of colonic tumors in Min mice by magnetic resonance micro-imaging. AB - We applied MRI to the in vivo detection of spontaneous colorectal tumors in a unique mouse model, the Fox Chase Cancer Center (FCCC) ApcMIN mouse. Unlike other Min (multiple intestinal neoplasia) strains, FCCC ApcMIN animals develop an appreciable number of tumors in the large intestine, which makes them an appropriate mouse model for colon cancer in humans. We describe a method for marking the colon on MRI data sets that involves a bowel-cleansing procedure and the insertion of a polyurethane tube (filled with an MRI contrast agent) fully into the colon. We found that tumors as small as 1.5 mm in diameter can be consistently identified from MRI datasets with a voxel size of 0.1 mm x 0.133 mm x 0.133 mm. Tumor volumes were determined from the MRM data sets with the use of a novel approach to planimetry in 3D data sets. We observed a correlation between tumor volume (as measured from the MRI datasets) and tumor weight of 0.942, and a P-value of 0.008, based on Spearman's test. These data show that MRI can be used to accurately monitor tumor growth in mouse models of colorectal carcinogenesis. PMID- 15334571 TI - In vivo diffusion tensor imaging of the human prostate. AB - This study demonstrates the feasibility of in vivo prostate diffusion tensor imaging (DTI) in human subjects. We implemented an EPI-based diffusion-weighted (DW) sequence with seven-direction diffusion gradient sensitization, and acquired DT images from six subjects using cardiac gating with a phased-array prostate surface coil operating in a linear mode. We calculated two indices to quantify diffusion anisotropy. The direction of the eigenvector corresponding to the leading eigenvalue was displayed by means of a color-coding scheme. The average diffusion values of the prostate peripheral zone (PZ) and central gland (CG) were 1.95 +/- 0.08 x 10(-3) mm2 s and 1.53 +/- 0.34 x 10(-3) mm2 s, respectively. The average fractional anisotropy (FA) values for the PZ and CG were 0.46 +/- 0.04 and 0.40 +/- 0.08, respectively. The diffusion ellipsoid in prostate tissue was anisotropic and approximated a prolate model, as shown in the color maps of the anisotropy. Consistent with the tissue architecture, the prostate fiber orientations were predominantly in the superior-inferior (SI) direction for both the PZ and CG. This study shows the feasibility of in vivo DTI and establishes normative DT values for six subjects. PMID- 15334572 TI - Visualization of beta-amyloid plaques in a transgenic mouse model of Alzheimer's disease using MR microscopy without contrast reagents. AB - The visualization of beta-amyloid plaque deposition in brain, a key feature of Alzheimer's disease (AD), is important for the evaluation of disease progression and the efficacy of therapeutic interventions. In this study, beta-amyloid plaques in the PS/APP transgenic mouse brain, a model of human AD pathology, were detected using MR microscopy without contrast reagents. beta-Amyloid plaques were clearly visible in the cortex, thalamus, and hippocampus of fixed brains of PS/APP mice. The distribution of plaques identified by MRI was in excellent agreement with those found in the immunohistological analysis of the same brain sections. It was also demonstrated that image contrast for beta-amyloid plaques was present in freshly excised nonfixed brains. Furthermore, the detection of beta-amyloid plaques was achieved with a scan time as short as 2 hr, approaching the scan time considered reasonable for in vivo imaging. PMID- 15334573 TI - Time course of 23Na signal intensity after myocardial infarction in humans. AB - Experimental studies demonstrated persistently increased 23Na content in nonviable myocardium post-myocardial infarction (MI). We hypothesized that nonviable myocardium in humans would show elevated 23Na content at all stages of infarct development, and therefore could be imaged with 23Na MRI. Ten patients were examined on days 4, 14, and 90 after infarction, and five of these patients participated in a 12-month follow-up. Double angulated short-axis cardiac 23Na images were obtained with the use of a 23Na surface coil and an ECG-triggered, 3D gradient-echo sequence. 1H T2-weighted imaging (N = 9) was performed on days 4, 14, and 90. Wall motion was assessed by cine MRI, and the infarct size was determined by late enhancement on day 90. The 23Na signal intensity (SI) of infarcted myocardium was expressed as the percentage increase over 23Na SI of noninfarcted myocardium. All of the patients showed an area of elevated SI on 23Na and 1H T2-weighted images that correlated with wall motion abnormalities and late enhancement. 23Na SI was highest on day 4. It then decreased until day 90, but remained elevated (39% +/- 18%, 31% +/- 17%, 28% +/- 13% on days 4, 14, and 90, respectively, P = 0.001). No further decrease was found 1 year after infarction (25% +/- 7%, P = 0.89 vs. day 90). 1H T2-weighted SI decreased between days 4 and 14, but on day 90 only six of nine patients had a residual elevated SI. Thus, 23Na SI is elevated in nonviable infarction at all time points following MI, and 23Na MRI may become a suitable technique for imaging nonviable myocardium in humans. PMID- 15334574 TI - Fat fractions and spectral T2 values in vertebral bone marrow in HIV- and non-HIV infected men: a 1H spectroscopic imaging study. AB - Fat fractions and spectral T2 values of fat and water within the vertebral marrow of non-HIV- and HIV-infected men were measured with the use of a Carr-Purcell Meiboom-Gill (CPMG) line scan spectroscopic imaging sequence. The fat fraction for the HIV-infected men (0.29 +/- 0.08) was significantly lower (P < 0.05, Student's unpaired t-test) than the fat fraction found in non-HIV-infected men (0.40 +/- 0.12). The mean water and fat T2 values did not differ between the two groups, and did not show any systematic dependence on fat fraction over the wide range of fat fractions encountered in this study. The marrow water and fat T2 values measured with the CPMG approach were markedly longer than the spectral T2 values reported by other groups using the more common point-resolved spectroscopy (PRESS) and stimulated-echo acquisition mode (STEAM) acquisitions. Proton spectroscopic studies of vertebral marrow revealed differences between non-HIV- and HIV-infected men that may prove useful for studying the effects of this disease and/or antiretroviral agents on body composition. PMID- 15334575 TI - Analysis of noise effects on DTI-based tractography using the brute-force and multi-ROI approach. AB - Diffusion tensor tractography based on line propagation is a promising and widely used technique, but it is known to be sensitive to noise and the size and location of the seed regions of interest (ROIs). The effects of these parameters on the tractography results were analyzed quantitatively using high-resolution diffusion tensor imaging (DTI) with a high signal-to-noise ratio (SNR) on a fixed mouse brain. The anterior commissure (AC), as judged from a T2-weighted image, was used as an anatomical reference within which the tracts could be located. Monte Carlo simulation was performed by adding Gaussian noise to the time domain data and repeating the tractography. Deviations of the tracking results were measured as a function of SNR. Such noise effects were evaluated for a simple one ROI approach and a combined two-ROI and brute-force (BF) approach. The influence of ROI size and location for the two-ROI + BF approach was also analyzed. The results confirmed the hypothesis that one can increase the validity of DTI-based tractography by adopting the BF and multi-ROI approach, with respect to the simple one-ROI approach. PMID- 15334576 TI - High-pass-low-pass (HP-LP) reconstruction of CINE phase-contrast MRI. AB - A new interpretation of CINE phase-contrast (PC) MRI is presented in which flowing blood is shown to induce separate high- and low-temporal-frequency components in the resulting image sequence. The flow velocities can then be extracted from the ratio of these two components, independently of any unknown phase offset. This interpretation leads to new insights into improving temporal resolution, eliminating noise, and reducing acquisition time in PC imaging. A specific example explored in this article uses a technique related to unaliasing by Fourier-encoding the overlaps using the temporal dimension (UNFOLD) to acquire PC velocity measurements at 54-68 time points in the cardiac cycle, all within a single breath-hold of <20 s. In experimental studies, these techniques were shown to yield improved signal-to-noise ratios (SNRs) in a flow phantom and cerebrospinal fluid (CSF) flow studies, and to resolve the formation of a flow vortex during left ventricular (LV) filling in humans. PMID- 15334577 TI - Semiquantitative fast flow velocity measurements using catheter coils with a limited sensitivity profile. AB - Flow measurements can be used to quantify blood flow during MR-guided intravascular interventional procedures. In this study, a fast flow measurement technique is proposed that quantifies flow velocities in the vicinity of a small RF coil attached to an intravascular catheter. Since the small RF coil receives signal from only a limited volume around the catheter, a spatially nonselective signal reception is employed. To enhance signal from flowing blood, and suppress unwanted signal contributions from static material, a slice-selective RF excitation is used. At a velocity sensitivity of 150 cm/s, a temporal resolution of 2 x TR = 10.2 ms can be achieved. The flow measurement is combined with an automatic slice positioning to facilitate measurements during interventional procedures. The influence of the catheter position in the blood vessel on the velocity measurement was analyzed in simulations. For blood vessels with laminar flow, the simulation showed a systematic deviation between catheter measurement and true flow between -15% and 80%. In four animal experiments, the catheter velocity measurement was compared with results from a conventional ECG-triggered 2D phase-contrast (PC) technique. The shapes of the velocity time curves in the abdominal aorta were nearly identical to the conventional measurements. A relative scaling factor of 0.69-1.19 was found between the catheter velocity measurement and the reference measurement, which could be partly explained by the simulation results. PMID- 15334578 TI - Assessing DTI data quality using bootstrap analysis. AB - Diffusion tensor imaging (DTI) is an established method for characterizing and quantifying ultrastructural brain tissue properties. However, DTI-derived variables are affected by various sources of signal uncertainty. The goal of this study was to establish an objective quality measure for DTI based on the nonparametric bootstrap methodology. The confidence intervals (CIs) of white matter (WM) fractional anisotropy (FA) and Clinear were determined by bootstrap analysis and submitted to histogram analysis. The effects of artificial noising and edge-preserving smoothing, as well as enhanced and reduced motion were studied in healthy volunteers. Gender and age effects on data quality as potential confounds in group comparison studies were analyzed. Additional noising showed a detrimental effect on the mean, peak position, and height of the respective CIs at 10% of the original background noise. Inverse changes reflected data improvement induced by edge-preserving smoothing. Motion-dependent impairment was also well depicted by bootstrap-derived parameters. Moreover, there was a significant gender effect, with females displaying less dispersion (attributable to elevated SNR). In conclusion, the bootstrap procedure is a useful tool for assessing DTI data quality. It is sensitive to both noise and motion effects, and may help to exclude confounding effects in group comparisons. PMID- 15334579 TI - Variable-rate selective excitation for rapid MRI sequences. AB - Balanced steady-state free precession (SSFP) imaging sequences require short repetition times (TRs) to avoid off-resonance artifacts. The use of slab selective excitations is common, as this can improve imaging speed by limiting the field of view (FOV). However, the necessarily short-duration excitations have poor slab profiles. This results in unusable slices at the slab edge due to significant flip-angle variations or aliasing in the slab direction. Variable rate selective excitation (VERSE) is a technique by which a time-varying gradient waveform is combined with a modified RF waveform to provide the same excitation profile with different RF power and duration characteristics. With the use of VERSE, it is possible to design short-duration pulses with dramatically improved slab profiles. These pulses achieve high flip angles with only minor off resonance sensitivity, while meeting SAR limits at 1.5 T. The improved slab profiles will enable more rapid 3D imaging of limited volumes, with more consistent image contrast across the excited slab. PMID- 15334580 TI - Flow-gated phase-contrast MRI using radial acquisitions. AB - Phase-contrast (PC) MRI is a noninvasive method for imaging blood and tissue velocity. PC experiments are typically segmented into many cardiac cycles in order to obtain sufficient spatial and temporal resolution, and thus require a gating signal to properly combine the data from multiple cardiac cycles. Although the surface electrode ECG is the gating waveform of choice for cardiovascular MRI, alternative gating methods are required for fetal cardiac imaging (due to the unavailability of fetal ECG signals) and for cases of ECG contamination via the strong magnetic fields within the MR scanner. This study demonstrates the feasibility of using imaging data from flow-encoded MRI experiments to derive times for cardiac gating. An undersampled radial k-space acquisition method is used to measure an image-derived flow-gating waveform in real-time, from which the gating times are derived. These gating times are used to reconstruct a conventional gated-segmented image series by combining the real-time data from multiple heartbeats. Flow-gated PC experiments were performed on five normal volunteers with slice prescriptions in four anatomic regions. The standard deviation (SD) of the difference between the flow-gating and ECG gating times ranged from 5 ms to 12 ms. PMID- 15334581 TI - Pulsatile motion effects on 3D magnetic resonance angiography: implications for evaluating carotid artery stenoses. AB - In-plane carotid artery motion during a 3D MR angiography (MRA) scan can significantly degrade the resulting image resolution. This study characterizes the effect of cardiac pulsatility on 3D contrast-enhanced (CE) MRA with elliptical centric acquisitions using a point-spread function (PSF) analysis. Internal carotid artery (ICA) motion was collected from volunteers and patients using both MR and ultrasound (US) scans. After measuring the carotid artery motion displacement, a simulation was performed which calculated the blurring effects for three different protocols: nongated and two different cardiac gating schemes. The motion sensitivity of each protocol was evaluated for different spatial resolutions. The selection of optimal imaging parameters for a given scan time was investigated. The final results showed that cardiac-gated acquisitions only over a limited region of k-space high spatial frequencies are more time efficient than cardiac gating for the entire k-space, as it allows for higher resolutions to be achieved and for capturing the arterial phase with low spatial frequencies. Selecting the optimal gating parameters depends directly on the motion characteristics of each individual. Our initial clinical experience is presented, and the need for a real-time tool that characterizes motion behavior for each individual as a prescan protocol is discussed. PMID- 15334582 TI - Susceptibility weighted imaging (SWI). AB - Susceptibility differences between tissues can be utilized as a new type of contrast in MRI that is different from spin density, T1-, or T2-weighted imaging. Signals from substances with different magnetic susceptibilities compared to their neighboring tissue will become out of phase with these tissues at sufficiently long echo times (TEs). Thus, phase imaging offers a means of enhancing contrast in MRI. Specifically, the phase images themselves can provide excellent contrast between gray matter (GM) and white matter (WM), iron-laden tissues, venous blood vessels, and other tissues with susceptibilities that are different from the background tissue. Also, for the first time, projection phase images are shown to demonstrate tissue (vessel) continuity. In this work, the best approach for combining magnitude and phase images is discussed. The phase images are high-pass-filtered and then transformed to a special phase mask that varies in amplitude between zero and unity. This mask is multiplied a few times into the original magnitude image to create enhanced contrast between tissues with different susceptibilities. For this reason, this method is referred to as susceptibility-weighted imaging (SWI). Mathematical arguments are presented to determine the number of phase mask multiplications that should take place. Examples are given for enhancing GM/WM contrast and water/fat contrast, identifying brain iron, and visualizing veins in the brain. PMID- 15334583 TI - Shim design using a linear programming algorithm. AB - The advent of open magnetic resonance imaging (MRI) scanners and dedicated MRI scanners tailored to specific body parts has led to an increasing number of noncylindrical MRI scanner geometries, for which noncylindrical gradients and shims are needed. These new scanner geometries are driving the need for fast, flexible shim design methods that can design shim coils for any geometry. A linear programming (LP) algorithm was developed to design minimum-power resistive shim coils on an arbitrary surface. These coils can be designed to produce any order shim field over an arbitrarily shaped target region, which can be placed anywhere within the coil. The resulting designs are relatively sparse and can be readily constructed. This algorithm was used to design and construct a seven-coil cylindrical shim set for a knee imaging magnet with a cylindrical homogeneous region. The algorithm was then used to design shim coils for a biradial head imager with an asymmetrically located spherical target region for brain imaging. PMID- 15334584 TI - Effects of inductive coupling on parallel MR image reconstructions. AB - Theoretical arguments and experimental results are presented that characterize the impact of inductive coupling on the performance of parallel MRI reconstructions. A simple model of MR signal and noise reception suggests that the intrinsic amount of spatial information available from a given coil array is unchanged in the presence of inductive coupling, as long as the sample remains the dominant source of noise for the coupled array. Any loss of distinctness in the measured coil sensitivities is compensated by information stored in the measured noise correlations. Adjustments to the theory are described to account for preamplifier noise contributions. Results are presented from an experimental system in which preamplifier input impedances are systematically adjusted in order to vary the level of coupling between array elements. Parallel image reconstructions using an array with four different levels of coupling and an acceleration factor up to six show average SNR changes of -7.6% to +7.5%. The modest changes in overall SNR are accompanied by similarly small changes in g factor. These initial results suggest that moderate amounts of inductive coupling should not have a prohibitive effect on the use of a given coil array for parallel MRI. PMID- 15334585 TI - Evaluation of internal MRI coils using ultimate intrinsic SNR. AB - The upper bounds of the signal-to-noise ratio (also known as the "ultimate intrinsic signal-to-noise ratio" (UISNR)) for internal and external coils were calculated. In the calculation, the body was modeled as a dielectric cylinder with a small coaxial cylindrical cavity in which internal coils could be placed. The calculated UISNR values can be used as reference solutions to evaluate the performance of internal MRI coils. As examples, we evaluated the performance of a loopless antenna and an endourethral coil design by comparing their ISNR with the UISNR. PMID- 15334586 TI - In vivo measurement and imaging of tumor oxygenation using coembedded paramagnetic particulates. AB - Tumor tissue oxygenation is an important parameter that is positively correlated to the chemo- or radiation treatment outcome of certain tumors. Hence, methods to accurately and noninvasively determine the concentration of oxygen (pO2) in tumors will be valuable. In this study, electron paramagnetic resonance (EPR) spectroscopy, utilizing microcrystalline particulates of lithium phthalocyanine (LiPc), was used to perform repeated measurements of pO2 as a function of tumor growth. We permanently embedded the particulates in the tumor by coimplanting them with RIF-1 tumor cells during inoculation in mice. This procedure enabled repeated measurements of oxygen concentration in the tumor to be obtained for >2 weeks during its growth phase. The particulates were stable and nontoxic to the tumor cells. Both an in vitro clonogenic assay and an in vivo tumor growth rate examination in C3H mice showed no apparent effect on cell proliferation or tumor growth rate. The measurements indicated that the pO2 of the tumor decreased exponentially with tumor growth and reached hypoxic levels ( approximately 4 mmHg) within 4 days after inoculation of the tumor cells. Spatial EPR imaging revealed a nonuniform distribution of the embedded particulates, which were localized mainly in the middle of the tumor volume. Oxygen mapping of the tumor, obtained by spectroscopic EPR imaging, showed significant variation of pO2 within the tumor. In summary, EPR spectroscopy and imaging with an embedded oximetry probe enabled accurate and repeated measurements of pO2 to be obtained in growing tumors under nonperturbing conditions. PMID- 15334587 TI - 2D J-resolved spiral spectroscopic imaging at 7 T: application to mobile lipid mapping in a rat glioma. AB - Lactate (Lac) and mobile lipids (Lip), which are present in rat gliomas, are difficult to map because their 1H resonances overlap in the 1.3 ppm region. 2D J resolved spectroscopy enables proper separation of the two resonance lines. To obtain high-spatial-resolution mapping of Lac and Lip resonances within a reasonable experiment time, we coupled 2D J-resolved spectroscopy with a fast spectroscopic imaging (SI) method, based on an out-and-in spiral k-space description. The method was applied to a rat glioma at 7 T, and Lac and Lip maps were reconstructed. The duration of SI (2D spatial, 2D spectral) was 64 min for a theoretical in-plane resolution of 1 x 1 mm, and a slice thickness of 2 mm (voxel size 8.2 microl, taking into account the point-spread function (PSF)). PMID- 15334588 TI - 31P MAS-NMR of human erythrocytes: independence of cell volume from angular velocity. AB - 31P magic angle spinning NMR (MAS-NMR) spectra were obtained from suspensions of human red blood cells (RBCs) that contained the cell-volume-sensitive probe molecule, dimethyl methylphosphonate (DMMP). A mathematical representation of the spectral-peak shape, including the separation and width-at-half-height in the 31P NMR spectra, as a function of rotor speed, enabled us to explore the extent to which a change in cell volume would be reflected in the spectra if it occurred. We concluded that a fractional volume change in excess of 3% would have been detected by our experiments. Thus, the experiments indicated that the mean cell volume did not change by this amount even at the highest spinning rate of 7 kHz. The mean cell volume and intracellular 31P line-width were independent of the packing density of the cells and of the initial cell volume. The relationship of these conclusions to other non-NMR studies of pressure effects on cells is noted. PMID- 15334589 TI - Persistent contrast enhancement by sterically stabilized paramagnetic liposomes in murine melanoma. AB - In the present research, we investigated the use of paramagnetic liposomes as contrast agents (CAs) for the detection of solid tumors. The liposomes were sterically stabilized by a polyethylene glycol (PEG) coating, and their size was constrained to approximately 100 nm. Dimyristoyl-sn-glycero-3-phosphoethanolamine N-diethylene-triaminepentaacetate (DMPE-DTPA) was used as the gadolinium-carrying fatty acid chain. The relaxation properties were characterized through nuclear magnetic relaxation dispersion (NMRD) measurements, and analyzed with the use of theories and computer programs that are adequate for slowly rotating systems. Their relaxivity at 1.5 T was found to be acceptable for in vivo use. We then tested the liposomes against B16-F10 murine melanomas using standard T1-weighted schemes at 1.5 T, and concentrations corresponding to 0.03 mmol/kg of gadolinium (i.e., three to six times lower than the concentration of the small gadolinium complexes in clinical use). The blood half-life was found to be 120 +/- 20 min. The experiments show a good contrast enhancement in the tumor (33% +/- 22%) 2 hr after administration, a further increase (43 +/- 27%) 20 hr after administration, and a decrease (25% +/- 14%) 54 hr after administration. High persistence of the CA was also observed in the liver and intestine, as expected in a hepatobiliar excretion pathway. PMID- 15334590 TI - Comparison between 2D and 3D gradient-echo sequences for MRI of human lung ventilation with hyperpolarized 3He. AB - Images of hyperpolarized 3He were acquired during breath-hold in four healthy volunteers with the use of an optimized 3D gradient-echo sequence. The images were compared with existing 2D gradient-echo methods. The average SNR from a 13 mm-thick slice in the peripheral lung was 1.4 times greater with 3D. In the airways the average SNR was 1.7 times greater with 3D. The higher SNR of 3D was particularly evident when regions of unimpeded gas diffusion, such as the major airways, were imaged with thin slices. This is because diffusion dephasing due to the slice-encoding gradient is minimized with a 3D sequence. The in vivo experimental findings were substantiated with experiments on phantoms of free gas, which showed more than four times the SNR with 3D compared to 2D. Theoretical simulations of the 2D and 3D k-space filters were also performed to predict the SNR and spatial resolution observed in the experimental images. PMID- 15334591 TI - Determining the longitudinal relaxation time (T1) of blood at 3.0 Tesla. AB - It is important to determine the longitudinal relaxation time of blood for black blood imaging, as well as for quantifying blood flow by arterial spin labeling (ASL). In this study a circulation system was used to measure blood T1 under physiological conditions at the new clinical field strength of 3.0T. It was found that 1/T1 in s(-1) was linearly dependent (P < 0.05) on hematocrit (Hct) within a normal range of 0.38-0.46. The relationships were 1/T1 = (0.52 +/- 0.15). Hct + (0.38 +/- 0.06) and 1/T1 = (0.83 +/- 0.07). Hct + (0.28 +/- 0.03) for arterial (oxygenation = 92% +/- 7%) and venous blood (69% +/- 8%), respectively, which led to estimated T1 values of 1664 +/- 14 ms (arterial) and 1584 +/- 5 ms (venous) at a typical human Hct of 0.42. The temperature dependencies of blood T1 were 22.3 +/- 0.6 ms/ degrees C and 19.8 +/- 0.8 ms/ degrees C for Hct values of 0.42 and 0.38, respectively. When a head coil transmit/receive setup was used, radiation damping caused a slight reduction (19 ms) of the measured T1 values. PMID- 15334592 TI - System for prostate brachytherapy and biopsy in a standard 1.5 T MRI scanner. AB - A technique for transperineal high-dose-rate (HDR) prostate brachytherapy and needle biopsy in a standard 1.5 T MRI scanner is demonstrated. In each of eight procedures (in four patients with intermediate to high risk localized prostate cancer), four MRI-guided transperineal prostate biopsies were obtained followed by placement of 14-15 hollow transperineal catheters for HDR brachytherapy. Mean needle-placement accuracy was 2.1 mm, 95% of needle-placement errors were less than 4.0 mm, and the maximum needle-placement error was 4.4 mm. In addition to guiding the placement of biopsy needles and brachytherapy catheters, MR images were also used for brachytherapy treatment planning and optimization. Because 1.5 T MR images are directly acquired during the interventional procedure, dependence on deformable registration is reduced and online image quality is maximized. PMID- 15334593 TI - Self-calibrated spiral SENSE. AB - Current standard sensitivity-encoded parallel imaging (SENSE) utilizes a fully sampled low-resolution reference scan to estimate the coil sensitivities. This reference scan adds scan time and may introduce misregistration artifacts. The purpose of this study was to investigate the feasibility of estimating the coil sensitivities for spiral SENSE directly from an undersampled k-space center. The limited spatial frequencies of the coil sensitivities, and the undersampling beyond the Nyquist radius cause image artifacts. A point spread function (PSF) analysis and experiments on both phantoms and humans identified an optimal radius for the k-space center by minimizing these image artifacts. The preliminary data indicate that self-calibrated SENSE is as accurate as standard SENSE, which uses a fully sampled reference scan. PMID- 15334594 TI - Catheter tracking and visualization using 19F nuclear magnetic resonance. AB - This work presents an investigation into catheter visualization and localization using 19F nuclear magnetic resonance (NMR) in conjunction with proton imaging. For this purpose, the imaging capabilities of a standard system were extended to allow for 19F excitation and signal detection. Two modes of operation were implemented: 1) a real-time tracking mode that provides tip tracking and automatic slice position updates interleaved with real-time, interactive proton imaging; and 2) a non-real-time catheter length visualization mode in which the entire length of a catheter can be assessed. Initial phantom experiments were conducted with the use of an angiographic balloon catheter filled with the blood substitute perfluorooctylbromide (PFOB). Using limited bandwidth excitation centered at the resonances of the CF2 groups of PFOB, we found that sufficient signal could be received to facilitate tip tracking during catheter motion and length visualization for various catheter configurations. The present approach is considered a promising alternative to existing methods, which either are associated with safety concerns (if active markers are employed) or suffer from insufficient, direction-dependent contrast (if passive visualization is used). Furthermore, our approach enables visualization of the entire length of the catheter. The proposed method provides a safe technique that, unlike electrical or optical devices, does not require modification of commercially available catheters. PMID- 15334596 TI - Acute and persistent protein synthesis inhibition following cerebral reperfusion. AB - Lack of recovery from protein synthesis inhibition (PSI) closely correlates with neuronal death following brain ischemia and reperfusion. It has therefore been suggested that understanding the mechanisms of PSI will shed light on the mechanisms of selective neuronal death following ischemia and reperfusion. It is now known that the PKR-like ER kinase (PERK)-mediated phosphorylation of eukaryotic initiation factor 2alpha (eIF2alpha) causes translation inhibition at initial reperfusion. Activation of PERK, in turn, indicates endoplasmic reticulum stress and activation of the unfolded protein response. However, phosphorylation of eIF2alpha is a transient event and can account for PSI only in the initial hours of reperfusion. Although a number of other regulators of protein synthesis, such as eIF4F, 4EBP-1, eEF-2, and S6 kinase, have been assessed following cerebral ischemia and reperfusion, the causes of prolonged PSI have yet to be fully elucidated. The purpose of the present article is to bring together the evidence indicating that, at minimum, postischemic PSI should be conceptualized as consisting of two components: an acute, transient component mediated by unfolded protein response-induced eIF2alpha phosphorylation and a longer term component that correlates with neuronal death. Ischemic tolerance appears to separate the acute and persistent components of reperfusion-induced translation inhibition. Specific models of the relationship among acute PSI, persistent PSI, and neuronal death are presented to clarify issues that have emerged from ongoing work in this area. PMID- 15334597 TI - The Mts1/S100A4 protein is a neuroprotectant. AB - Mts1 (S100A4) is a calcium-binding protein of the EF-hand type, belonging to the S100 family of proteins. The mts1/S100A4 gene was originally isolated from tumor cell lines, and the protein is believed to play an important role in tumor progression. More recently, oligomeric, but not dimeric, forms of Mts1 have been shown to have a neuritogenic effect when added extracellularly to hippocampal neurons. Here we show increased neurite outgrowth in two other cell types, dopaminergic and cerebellar neurons, in response to treatment with Mts1 oligomers. Moreover, we demonstrate that Mts1 acts as a neuroprotectant in primary cerebellar, dopaminergic, and hippocampal neurons induced to undergo cell death. Interestingly, the survival of the cerebellar and hippocampal neurons increased as a result of treatment with Mts1 not only in oligomeric form but also -although to a lesser extent--in dimeric form. The inhibition of death in cerebellar neurons by Mts1 was accompanied by an inhibition of DNA fragmentation, but Mts1 did not affect the activity of caspases-3 and -6. In hippocampal neurons, cell death induced by the amyloid-beta peptide (Abeta(25-35)) was characterized by an increase in caspase-3 and -6 activity, but no DNA fragmentation was observed. As in cerebellar neurons, the induced increase in caspase activity in hippocampal neurons was not affected by Mts1. PMID- 15334598 TI - Differential roles of fibroblast growth factor-2 during development and maintenance of auditory sensory epithelia. AB - Fibroblast growth factor-2 (FGF2) has been postulated to be a key regulator involved in the proliferation, differentiation, and regeneration of sensory hair cells. Here we have addressed the potential functions of FGF2 during the formation and regeneration of the auditory epithelium in chicken and mice. By using viral gene transfer, based on herpes simplex type 1 virus (HSV-1), we show that ectopically applied FGF2 drastically increases the number of cells expressing early hair cell markers during embryonic development in avians. Intriguingly, FGF2 does not stimulate cell division during this process. These data suggest that FGF2 plays a role during differentiation of sensory hair cells in avians. To address the potential functions of FGF2 during murine inner ear development, we analyzed FGF2 mouse mutants. Mice lacking FGF2 showed normal formation of the inner ear, and no abnormalities were observed at the adult stage. Moreover, FGF2 mouse mutants showed similar hearing thresholds compared with those observed in control mice before and after noise damage. Therefore, endogenous FGF2 appears not to be essential for the development or functional maintenance of the auditory organ in mammals. In light of these results, the differential roles of FGF2 in the vertebrate inner ear are discussed with respect to its previously postulated functions. PMID- 15334599 TI - Fine structure of degeneration in the cochlear nucleus of the chinchilla after acoustic overstimulation. AB - To study plastic changes in the cochlear nucleus after acoustic stimulation, adult chinchillas were exposed once to a 4-kHz octave-band noise at 108 dB SPL for 3 hr. After survival times of 1, 2, 4, 8, and 16 weeks, samples were taken for electron microscopy from a part of the cochlear nucleus, where cochlear nerve fibers degenerated after the noise exposure. Progressive changes in fine structure were characterized as early, intermediate, and late stages of degeneration. Freshly occurring synaptic degeneration appeared in each period from 1-16 weeks. Endings with large round vesicles, putative excitatory synapses of the cochlear nerve, displayed progressive increases in neurofilaments and enlarged synaptic vesicles. Compared to controls, synaptic vesicles seemed fewer, often in small clusters in the interior of endings, and smaller in the synaptic zone. These early changes progressed to mitochondrial disintegration and overt "watery" degeneration. Some surviving endings, however, were shrunken and displaced partially by enlarged spaces in the synaptic complex. Dense-cored vesicles gathered in these endings. In terminals with pleomorphic and flattened vesicles, presumed inhibitory endings, cytological changes appeared within 1 week and persisted for months. The synaptic endings darkened, some vesicles disintegrated, and many smaller flatter vesicles collapsed into heaps. Especially at the presynaptic membrane, vesicles were shriveled, but a few mitochondria were preserved. Without overt signs of synaptic degeneration, some of these cytological changes presumably reflect reduced synaptic activity in the inhibitory endings. These changes may contribute to a continuing process associated with abnormal auditory functions, including hyperacusis and tinnitus. PMID- 15334600 TI - Fine structure of long-term changes in the cochlear nucleus after acoustic overstimulation: chronic degeneration and new growth of synaptic endings. AB - The companion study showed that acoustic overstimulation of adult chinchillas, with a noise level sufficient to damage the cochlea, led to cytological changes and degeneration of synaptic endings in the cochlear nucleus within 1-16 weeks. In the present study, the same stimulus was used to study the long-term effects on the fine structure of synaptic endings in the cochlear nucleus. For periods of 6 and 8 months after a single exposure to a damaging noise level, there ensued a chronic, continuing process of neurodegeneration involving excitatory and inhibitory synaptic endings. Electron microscopic observations demonstrated freshly occurring degeneration even as late as 8 months. Degeneration was widespread in the neuropil and included the synapses on the globular bushy cell, which forms part of the main ascending auditory pathway. Neurodegeneration was accompanied by newly formed synaptic endings, which repopulated some of the sites vacated previously by axosomatic endings on globular bushy cells. Many of these synaptic endings must arise from central interneurons. The findings suggest that overstimulation can induce a self-sustaining condition of progressive neurodegeneration accompanied by a new growth of synaptic endings. Noise-induced hearing loss thus may progress as a neurodegenerative disease with the capacity for synaptic reorganization within the cochlear nucleus. PMID- 15334601 TI - Quantitative study of degeneration and new growth of axons and synaptic endings in the chinchilla cochlear nucleus after acoustic overstimulation. AB - To determine if acoustic overstimulation altered synaptic connections in the cochlear nucleus, anesthetized adult chinchillas, with one ear protected by a silicone plug, were exposed for 3 hr to a 108-dB octave-band noise, centered at 4 kHz, and allowed to survive for periods up to 32 weeks. This exposure led to cochlear damage in the unprotected ear, mainly in the basal regions of the organ of Corti. The anterior part of the ipsilateral posteroventral cochlear nucleus consistently contained a band of degenerating axons and terminals, in which electron microscopic analysis revealed substantial losses of axons and synaptic terminals with excitatory and inhibitory cytology. The losses were significant after 1 week's survival and progressed for 16-24 weeks after exposure. By 24-32 weeks, a new growth of these structures produced a resurgence in the number of axons and terminals. The net number of excitatory endings fully recovered, but the quantity with inhibitory cytology was only partially recouped. Neuronal somata lost both excitatory and inhibitory endings at first and later recovered a full complement of excitatory but not inhibitory terminals. Dendrites suffered a net loss of both excitatory and inhibitory endings. Excitatory and inhibitory terminals with unidentified postsynaptic targets in the neuropil declined, then increased in number, with excitatory terminals exhibiting a greater recovery. These findings are consistent with a loss and regrowth of synaptic endings and with a reorganization of synaptic connections that favors excitation. PMID- 15334602 TI - Microarray analysis of acute and delayed gene expression profile in rats after focal ischemic brain injury and reperfusion. AB - Temporal changes in gene expression were measured using DNA microarrays after 30 min or 2-hr transient middle cerebral artery occlusion (MCAo) in rats. Total RNA was extracted from the injured hemisphere at 30 min, 4 hr, 8 hr, 24 hr, 3 days, and 7 days after MCAo for GeneChip analysis using Affymetrix U34 Rat Neurobiology arrays (1,322 functional genes). In total, 267 genes were expressed differentially: 166 genes were upregulated, 94 genes were downregulated, and 7 genes were biphasically up- and downregulated. Among all differentially expressed genes, 88 were newly identified as associated with ischemic brain injury. Most affected genes were distributed among 12 functional categories. Immediate early genes, transcription factors, and heat shock proteins were upregulated as early as 30 min after MCAo, followed by the upregulation of inflammation, apoptosis, cytoskeletal, and metabolism genes, which peaked within 4-24 hr of injury. Neurotrophic growth factors exhibited a sustained upregulation beginning 24 hr after MCAo and persisting through 7 days post-injury. Three classes of genes were downregulated with distinct temporal patterns: ion channel genes and neurotransmitter receptor genes were downregulated between 8-24 hr after injury, whereas synaptic proteins genes were downregulated between 3-7 days after MCAo. Downregulation of synaptic protein gene expression after ischemic injury is of particular interest because of its conspicuously delayed pattern as a functional group, which has not been reported previously and may play a role in post-injury recovery. PMID- 15334603 TI - Microarray analysis of postmortem temporal cortex from patients with schizophrenia. AB - To examine molecular mechanisms associated with schizophrenia this study measured expression of approximately 12,000 genes in the middle temporal gyrus from 12 subjects with schizophrenia and 14 matched normal controls. Among the most consistent changes in genes with robust expression were significant decreases in the expression of myelination-related genes MAG, PLLP (TM4SF11), PLP1, ERBB3 in subjects with schizophrenia. There was also altered expression of genes regulating neurodevelopment (TRAF4, Neurod1, histone deacetylase 3), a circadian pacemaker (PER1), and several other genes involved in regulation of chromatin function and signaling mechanisms. These findings support the hypothesis that schizophrenia is associated with abnormalities in oligodendroglia and provide initial evidence suggesting a role for epigenetic mechanisms and altered circadian rhythms in this disorder. PMID- 15334604 TI - Proteomic analysis of phosphotyrosyl proteins in the rat brain: effect of butorphanol dependence. AB - Butorphanol (17-cyclobutylmethyl-3,14-dihydroxymorphinan) tartrate (Stadol) is a mixed agonist-antagonist opioid analgesic agent that is about five to seven times as potent as morphine in analgesic effects. The chronic use of butorphanol produces physical dependence in humans and animals. Phosphorylation plays a very important role in developing butorphanol dependence; however, global phosphorylation events induced by chronic butorphanol administration have not been reported. The aim of this study is to determine the alteration of tyrosine phosphorylation of brain frontal cortical proteins in butorphanol-dependent rats using a proteomic approach. Dependence was produced by continuous intracerebroventricular (i.c.v.) infusion of butorphanol (26 nmol/microl/hr) for 72 hr via osmotic minipump in rats. Similar patterns of protein expression were detected by two-dimensional electrophoresis (2-DE) in brain frontal cortex of butorphanol-dependent and saline-treated control rats. All 65 phosphotyrosyl (p Tyr) protein spots detected in pH 3-10 phosphotyrosine 2-DE of control rat brains were detected in butorphanol-dependent rat brains. The densities of most p-Tyr protein spots were increased in butorphanol-dependent rat brains compared to saline-treated control samples. Eighteen additional p-Tyr protein spots were detected in pH 3-10 2-DE images of butorphanol-dependent rat brains. Immobilized pH strips with three different narrow pH ranges were examined to improve the resolution of p-Tyr proteins in 2-DE gels. Fifty-three p-Tyr protein spots were identified as known proteins involved in cell cytoskeleton, cell metabolism, and cell signaling. This proteomic approach can provide useful information for understanding the complex mechanism of butorphanol dependence in vivo. PMID- 15334605 TI - Acute treatment with MgSO4 attenuates long-term hippocampal tissue loss after brain trauma in the rat. AB - Previous studies have shown that magnesium salts and the noncompetitive N-methyl D-aspartate (NMDA) receptor antagonist, NPS 1506, attenuated short-term cognitive deficits and histopathological changes associated with traumatic brain injury (TBI). We evaluated the long-term effects of both therapies after brain trauma. Young adult rats were subjected to parasagittal fluid-percussion brain injury and received either MgSO(4) (125 micromol/400 g rat; n = 12) 15 min post-injury, NPS 1506 (1.15 mg/kg; n = 12) 15 min and 4 hr post-injury, or vehicle (n = 9) 15 min post-injury. Uninjured animals (sham) received vehicle (n = 10). Learning function in these animals was evaluated using a water maze paradigm 8 months after injury or sham treatment, and the brains were examined for cortical and hippocampal tissue loss. Compared to sham animals, injured vehicle-treated animals displayed a substantial learning dysfunction, indicated by an increased latency to find a hidden platform in the water maze (P < 0.001). No improvements in learning, however, were found for injured animals treated with NPS 1506 or MgSO(4). Injury induced >30% loss of tissue in the ipsilateral cortex in vehicle treated animals that was not reduced in animals treated with either NPS 1506 or MgSO(4). Treatment with MgSO(4) significantly reduced progressive tissue loss in the hippocampus (P < 0.001). These findings are the first to demonstrate long term neuroprotection of hippocampal tissue by an acute treatment in a TBI model. These data also show that the previously reported broad efficacy of MgSO(4) or NPS 1506 observed shortly after brain trauma could not be detected 8 months post injury. PMID- 15334606 TI - Modulation of CREB expression and phosphorylation in the rat nucleus accumbens during nicotine exposure and withdrawal. AB - The nucleus accumbens region of the brain has been shown to play a role in reward and reinforcing mechanisms of drugs of abuse. To understand the molecular mechanisms of nicotine addiction, the present investigation examined the effects of acute and chronic nicotine treatment and its withdrawal on cAMP-responsive element binding (CREB) protein expression and phosphorylation (serine-133) in nucleus accumbens (NAc) structures of rats. it was found that acute treatment (1 and 18 hr of withdrawal) with nicotine had no effects on total creb and phosphorylated CREB (p-CREB) protein levels in shell or core structures of rat NAc. On the other hand, 18-hr withdrawal after chronic nicotine exposure produced significant reductions in the total CREB and p-CREB protein levels in the shell but not in core structures of nac. interestingly, nicotine withdrawal (1 hr) after chronic exposure maintained normal levels of total CREB and p-CREB protein levels in the shell and core structures of NAc. These results suggest the possibility that decreased CREB activity in the shell of NAc may be associated with abnormal reward mechanisms during nicotine withdrawal after chronic exposure. PMID- 15334607 TI - Protective effect of green tea polyphenol EGCG against neuronal damage and brain edema after unilateral cerebral ischemia in gerbils. AB - Previous studies have demonstrated that a green tea polyphenol, (-) epigallocatechine gallate (EGCG), has a potent free radical scavenging and antioxidant effect. Glutamate leads to excitotoxicity and oxidative stress, which are important pathophysiologic responses to cerebral ischemia resulting in brain edema and neuronal damage. We investigated the effect of EGCG on excitotoxic neuronal damage in a culture system and the effect on brain edema formation and lesion after unilateral cerebral ischemia in gerbils. In vitro, excitotoxicity was induced by 24-hr incubation with N-methyl-D-aspartate (NMDA; 10 microM), AMPA (10 microM), or kainate (20 microM). EGCG (5 microM) was added to the culture media alone or with excitotoxins. We examined malondialdehyde (MDA) level and neuronal viability to evaluate the effect of EGCG. In vivo, unilateral cerebral ischemia was induced by occlusion of the right common carotid artery for 30, 60, or 90 min and followed by reperfusion of 24 hr. Brain edema, MDA, and infarction were examined to evaluate the protective effect of EGCG. EGCG (25 or 50 mg/kg, intraperitoneally) was administered twice, at 30 min before and immediately after ischemia. EGCG reduced excitotoxin-induced MDA production and neuronal damage in the culture system. In the in vivo study, treatment of gerbils with the lower EGCG dose failed to show neuroprotective effects; however, the higher EGCG dose attenuated the increase in MDA level caused by cerebral ischemia. EGCG also reduced the formation of postischemic brain edema and infarct volume. These results demonstrate EGCG may have future possibilities as a neuroprotective agent against excitotoxicity-related neurologic disorders such as brain ischemia. PMID- 15334608 TI - Temporal profile of ultrastructural changes in cortical neurons after a photochemical lesion. AB - A photochemical lesion was induced in the right sensory motor cortex of rat brains. We examined at various time points the occurrence of different types of neuronal death with respect to a potential therapeutic window. The lesion appearance was documented by magnetic resonance imaging, and functional recovery was evaluated by behavioral tests showing recovery at 48 hr after lesioning. At 0.5, 1, 3, 6, 12, 24, 48, and 72 hr postlesion, cortical layers IV and V were examined by light and electron microscopy. Ultrastructural changes, which corresponded well to light microscopy findings, were found in both hemispheres. In the lesioned area, the neuropil appeared disorganized at 0.5 hr, and apoptotic and necrotic cell death was found at 0.5-3 hr. After 3 hr, the tissue was disintegrated. On the contralateral side, chromatin clumping appeared at 0.5-3 hr. At 3 hr, ruptured membranes were found, a sign of irreversible cell death. At 6-72 hr, the membranes were intact, and the chromatin was not clumped but heterogeneously distributed. The nuclei contained dispersed nucleoli at 48-72 hr. The morphology correlated well with magnetic resonance images and functional behavior. Our study demonstrates that a photochemical lesion is a useful model for studying morphological changes in injured cells. It results in a permanent infarction within 3 hr. In that the morphology on the contralateral side drastically changed between 3 and 6 hr, the cellular alterations at these time points might represent a break point at which cells either progress toward cell death or recover. PMID- 15334609 TI - Gamma-aminobutyric acidergic interneuron vulnerability to aging in canine prefrontal cortex. AB - The aged dog is considered a promising model for examining molecular and cellular processes involved in a variety of human neurological disorders. By using the canine counterpart of senile dementia of the Alzheimer's type (ccSDAT), we investigated the specific vulnerability of the gamma-aminobutyric acid (GABA) cortical subset of interneurons, characterized by their calcium-binding protein content, to neuronal death. Dogs representing a large variety of breeds were classified into three groups: young control, aged control, and ccSDAT. In all dogs, the general distribution and cell typology of parvalbumin-, calretinin-, and calbindin-positive neurons were found to be similar to those in the human. As in Alzheimer's disease patients, neurons displaying parvalbumin or calretinin immunoreactivity were resistant and the calbindin-positive ones depleted. Together with aging, amyloid deposition in its early phase (stage II) participates in this specific neuronal death, but with a lower potency. In conclusion, our data provide evidence that preservation of GABAergic cortical interneurons has to be focused on the early stage of beta-amyloid deposition. We also demonstrate the usefulness of dogs of all breeds for investigating the early phases of human brain aging and Alzheimer's disease. PMID- 15334610 TI - Can topically applied optical clearing agents increase the epidermal damage threshold and enhance therapeutic efficacy? PMID- 15334611 TI - In vivo multiphoton fluorescence imaging: a novel approach to oral malignancy. AB - BACKGROUND AND OBJECTIVE: Current techniques for oral diagnosis require surgical biopsy of lesions, and may fail to detect early malignant change. Non-invasive, sensitive tools providing early detection of oral cancer and a better understanding of malignant change are needed. These studies evaluated in vivo multiphoton excited fluorescence (MPM) techniques to (1) map epithelial and subepithelial changes through out oral carcinogenesis and (2) serve as an effective diagnostic modality. STUDY DESIGN/MATERIALS AND METHODS: In the hamster model (n = 70), epithelial and subepithelial change was imaged in vivo throughout carcinogenesis. MPM- and histopathology-based diagnoses on a scale of 0 (healthy) 6 (squamous cell carcinoma [s.c.c.]) were scored by two pre-standardized investigators. RESULTS: Collagen matrix and fibers, cellular infiltrates, blood vessels, and microtumors were clearly visible. MPM agreed with the histopathology for 88.6% of diagnoses. CONCLUSIONS: In vivo MPM images provide (1) high resolution information on specific components of the carcinogenesis process (2) an excellent basis for oral diagnostics. PMID- 15334612 TI - Pulpal safety of 9.6 microm TEA CO2 laser used for caries prevention. AB - BACKGROUND AND OBJECTIVES: Lasers are used for several procedures involving hard and soft tissues of the oral cavity. A potential future application is the use of the CO2 laser to alter the surface structure of tooth enamel to render it more resistant to caries. A new 9.6 microm wavelength transverse excited atmospheric pressure (TEA) CO2 laser (Argus Photonics, Jupiter, FL) has been investigated as a device that can be used for this purpose without harming the dental pulp. STUDY DESIGN/MATERIALS AND METHODS: Erupted caries- and restoration-free third molars (n = 24 participants; 74 teeth) were used in the study. Teeth were irradiated at an incident fluence of 1.5 J/cm2, a repetition rate of 10 Hz and a spot size 1 mm in diameter. At the low and high settings, 200-400 pulses at 5-8 microseconds pulse duration were delivered at 12 mJ per pulse for a total energy of 2.4 or 4.8 J delivered for 20 or 40 seconds, respectively. Other teeth were subjected to a sham dental procedure (positive control) or no procedure (negative control). Prior to testing, radiographs were taken of all teeth, and they were assessed pulpally using heat, cold, and electricity to determine vitality. The teeth were removed either immediately or at 1 week or 1 month after testing. RESULTS: Teeth were bioprepared and examined histologically for signs of inflammation. Only one tooth developed symptoms of sensitivity to cold for 10 days following exposure to the high power level. The sensitivity was of fleeting duration and was judged to be reversible pulpitis. All teeth tested responded normally at pre-testing and pre-extraction time periods. Histological examination of all teeth disclosed no indication of an inflammatory response in the pulp tissue at any time point. All sections appeared normal with no changes seen in the normal pulpal morphology. CONCLUSIONS: We conclude that the 9.6 microm wavelength laser causes no permanent/serious pulpal damage at the energy levels used and can be used safely for caries prevention treatments in humans. PMID- 15334613 TI - Bactericidal effect of different laser systems in the deep layers of dentin. AB - BACKGROUND AND OBJECTIVES: In recent years, various laser systems have gained importance in the field of laser-assisted endodontics, namely the Nd:YAG, the diode, the Er:YAG, and the Er,Cr:YSGG laser. Individual studies have been carried out so far, focusing on the respective wavelength, its specific bactericidal capabilities, and potential usefulness is root-canal disinfection. The present in vitro investigation however, was performed to compare the microbicidal effect of these laser systems under standardized conditions and to draw a conclusion upon their relative effectiveness in the deep layers of dentin. STUDY DESIGN/MATERIALS AND METHODS: In total, 360 slices of root dentin with a thickness of 1 mm were obtained by longitudinal cuts of freshly extracted human premolars. The samples were steam sterilized and subsequently inoculated with a suspension of either Escherichia coli or Enterococcus faecalis. After the incubation, the samples were randomly assigned to the four different laser systems tested. Each laser group consisted of two different operational settings and a control. The dentinal samples underwent "indirect" laser irradiation through the dentin from the bacteria-free side and were then subjected to a classical quantitative microbiologic evaluation. To assess the temperature increase during the irradiation procedure, additional measurements were carried out using a thermocouple. RESULTS: Microbiology indicated that all laser systems were capable of significant reductions in both test strains. At an effective output power of 1 W, E. coli was reduced by at least three log steps in most of the samples by the tested wavelengths, with the best results for the Er:YAG laser showing complete eradication of E. coli in 75% of the samples. E. faecalis, a stubborn invader of the root canal, showed minor changes in bacterial count at 1 W. Using the higher setting of 1.5 W, significant reductions of E. coli were again observed with all laser systems, where only the diode and the Er:YAG laser were capable of complete eradication of E. faecalis to a significant extent. There was no significant relation between the temperature increase and the bactericidal effect. CONCLUSIONS: The present study demonstrates that all the wavelengths investigated are suitable for the disinfection of even the deeper layers of dentin and may prove to constitute valuable tools in state-of-the-art endodontics. PMID- 15334614 TI - Effects of low-intensity laser therapy on the orthodontic movement velocity of human teeth: a preliminary study. AB - BACKGROUND AND OBJECTIVES: Low-intensity laser therapy (LILT) has been studied in many fields of dentistry, but to our knowledge, this is the first time that its effects on orthodontic movement velocity in humans are investigated. STUDY DESIGN/PATIENTS AND METHODS: Eleven patients were recruited for this 2-month study. One half of the upper arcade was considered control group (CG) and received mechanical activation of the canine teeth every 30 days. The opposite half received the same mechanical activation and was also irradiated with a diode laser emitting light at 780 nm, during 10 seconds at 20 mW, 5 J/cm2, on 4 days of each month. Data of the biometrical progress of both groups were statistically compared. RESULTS: All patients showed significant higher acceleration of the retraction of canines on the side treated with LILT when compared to the control. CONCLUSIONS: Our findings suggest that LILT does accelerate human teeth movement and could therefore considerably shorten the whole treatment duration. PMID- 15334615 TI - Feasibility study of normal and septic tracheal imaging using optical coherence tomography. AB - BACKGROUND AND OBJECTIVES: Optical coherence tomography (OCT) is an imaging technology that may be adapted for use with flexible fiberoptic bronchoscopy, potentially allowing it to play an important role in pulmonary diagnostics. The goal of this study was to evaluate the feasibility of OCT to image tracheal pathology. STUDY DESIGN/MATERIALS AND METHODS: Tracheas were harvested from normal and septic New Zealand White rabbits and imaged using OCT. Two delivery devices were employed. One was a moving stage with an objective lens and collimator, the other a linear scanning flexible fiberoptic catheter using a GRIN lens and prism for endoscopic OCT. After OCT images were obtained from normal and septic tracheas, the excised tissues were prepared for standard histologic examination. Areas imaged by OCT were compared with corresponding histology slides. RESULTS: OCT images demonstrated in detail tracheal sub-surface structures such as the epithelium, lamina propria, submucosa, and cartilage. The appearance of structures imaged by OCT corresponded very well with histologic pictures obtained by light microscopy. The OCT images from septic tracheas showed marked swelling of the mucosal and submucosal layers. Such pathology was equally imaged by either the moving stage or fiberoptic catheter for endoscopic OCT. CONCLUSIONS: OCT images of the trachea can distinguish many sub-surface structural features usually requiring biopsy and light microscopy for visualization. Marked differences between normal and septic trachea were apparent in OCT images. In the future, OCT may be a valuable tool for evaluating tracheal pathology in situ with high image resolution. PMID- 15334616 TI - Lasers in pediatric airway surgery: current and future clinical applications. AB - BACKGROUND AND OBJECTIVES: The smaller anatomy and limited access to instrumentation pose a challenge to the pediatric airway surgeon. The enhanced precision and ability to photocoagulate tissue while operating with the laser enhances the surgeon's ability to successfully treat unique pediatric conditions. STUDY DESIGN/MATERIALS AND METHODS: A comprehensive literature review was performed using the Medline (1995-2004) database. RESULTS: Due to its shallow tissue penetration and thermal effect, the carbon dioxide (CO2) laser is generally considered the laser of choice for pediatric airway applications. The potential for increased scarring and damage to underlying tissue caused by the greater penetration depth and thermal effect of the Nd:YAG and potassium-titanyl phosphate (KTP) lasers preclude their widespread use in this population. CONCLUSIONS: In this review, we will discuss the clinical applications of lasers in pediatric airway surgery, the current state of the art, where the current technology is deficient, and the potential applications of novel laser systems. PMID- 15334617 TI - Clinical application of Nd:YAG laser for the treatment of small hepatocellular carcinoma with new shaped laser probe. AB - BACKGROUND AND OBJECTIVES: Surgical resection of hepatocellular carcinoma (HCC) is not always regarded as the first choice of treatment, because most HCCs develop in cirrhotic liver. It becomes adaptation of minimally invasive treatments for HCC. In animal experiments, we previously reported that our newly developed neodymium:yttrium aluminum garnet (Nd:YAG) laser system could ablate liver tissue in short periods. STUDY DESIGN/PATIENTS AND METHODS: Six HCC cases (mean diameter; 16.3 +/- 3.50 mm) were enrolled. The Nd:YAG laser generator with a wavelength of 1,064 nm at 10 W of power emitted the laser in a forward fashion and in all patients the laser beam safely irradiated the target lesion. RESULTS: There was no recurrence of HCC in the laser-ablated area in six patients for duration of 21.8+/- 4.35 months. A local recurrence was found in only one patient. CONCLUSIONS: This study showed the clinical usefulness of laser ablation in the treatment of HCC. PMID- 15334618 TI - Albumin-genipin solder for laser tissue repair. AB - BACKGROUND: Laser tissue soldering (LTS) is an alternative technique to suturing for tissue repair that avoids foreign body reaction and provides immediate sealing of the wound. One of the major drawbacks of LTS, however, is the weak tensile strength of the solder welds when compared to sutures. In this study, a crosslinking agent of low cytotoxicity was investigated for its ability to enhance the bond strength of albumin solders with sheep intestine. STUDY DESIGN/MATERIALS AND METHODS: Solder strips were welded onto rectangular sections of sheep small intestine using a diode laser. The laser delivered in continuous mode a power of 170 +/- 10 mW at lambda = 808 nm, through a multimode optical fiber (core size = 200 microm) to achieve a dose of 10.8 +/- 0.5 J/mg. The solder thickness and surface area were kept constant throughout the experiment (thickness = 0.15 +/- 0.01 mm, area = 12 +/- 1.2 mm2). The solder was composed of 62% bovine serum albumin (BSA), 0.38% genipin, 0.25% indocyanin green dye (IG), and water. Tissue welding was also performed with a BSA solder without genipin, as a control group. The repaired tissue was tested for tensile strength by a calibrated tensiometer. Murine fibroblasts were also cultured in extracted media from heat-denatured genipin solder to assess cell growth inhibition in a 48 hours period. RESULTS: The tensile strength of the genipin solder was doubled that of the BSA solder (0.21 +/- 0.04 N and 0.11 +/- 0.04 N, respectively; P = 10(-15) unpaired t-test, N = 30). Media extracted from crosslinked genipin solder showed negligible toxicity to fibroblast cells under the culture conditions examined here. CONCLUSION: Addition of a chemical crosslinking agent, such as genipin, significantly increased the tensile strength of adhesive-tissue bonds. A proposed mechanism for this enhanced bond strength is the synergistic action of mechanical adhesion with chemical crosslinking by genipin. PMID- 15334619 TI - Repetitive Er:YAG laser irradiation of human skin: a histological evaluation. AB - BACKGROUND AND OBJECTIVE: Deep coagulation of skin collagen by Er:YAG laser repetitive pulses has been predicted by previous theoretical models and later demonstrated on animal skin. The goal of this study was to determine the effect of repetitive Er:YAG laser pulses on human skin and its response to this treatment. STUDY DESIGN/MATERIALS AND METHODS: Lid skin of six female volunteers with blepharochalasis has been treated with laser at day 0, 7, and 21 before elective surgery-blepharoplasty. The treated skin was excised as part of the procedure and prepared for further histological examination. We used a 2,940 nm Er:YAG laser (Fidelis M320A by Fotona) with 'smooth' mode parameters: fluence from 0.50 to 2.00 J/cm2; six pulses per packet; 550 microsecond/pulse, 250 millisecond/packet; single pass, no overlapping; spot size 5 mm; repetition rate 20 Hz. RESULTS: We observed deep collagen denaturation at laser fluences of 1.25 J/cm2 and over; epidermal damage was proportional to fluence with total coagulation of the epidermal layer at fluences of 1.75 J/cm2 and over. At day 7 after laser treatment we observed a complete regeneration of the epidermal layer and a regeneration zone within the dermis with prominent infiltration of CD68+ monocytes/macrophages. At day 21 after laser treatment we observed collagen remodeling and (myo-)fibroblast proliferation at tissue depths of up to 240 microm. CONCLUSIONS: Repetitive Er:YAG laser irradiation is effective in deep denaturation and remodeling of human skin collagen in vivo, with less epidermal damage compared to standard Er:YAG laser skin resurfacing. PMID- 15334620 TI - Treatment of vitiligo by 308-nm excimer laser: an evaluation of variables affecting treatment response. AB - BACKGROUND AND OBJECTIVES: To determine the true efficacy of the 308-nm excimer laser for the treatment of vitiligo, while taking into account confounding factors such as anatomic site of treatment, age, sex, skin type, MED, and duration of evolution of the vitiligo. STUDY DESIGN/MATERIALS AND METHODS: Thirty five patients with vitiligo were included. Each lesion was treated twice a week by the 308-nm excimer laser for a maximum of 24 sessions. Efficacy was blindly evaluated by two independent physicians. RESULTS: Repigmentation was noted in 46 plaques/52 (88.5%). Repigmentation rate (75%) was obtained in 14 (26.9%). In "UV sensitive" areas (face, neck, trunk), 8/14 lesions (57.1%) had a repigmentation rate, 75% versus 6/38 (15.8%) in "UV resistant" areas (bony prominences and extremities) (P = 0.031). No relationship could be established between response to the treatment and the following variables: age, sex, skin type, MED, and duration of evolution of the vitiligo (respectively, P = 1, 0.666, 0.566, 0.628, 0.521). CONCLUSIONS: An aesthetically reasonable result is achieved essentially in "UV sensitive" areas, thus appearing to be the appropriate places of choice for this treatment. PMID- 15334621 TI - The effect of indocyanine green pretreatment on the parameters of transscleral diode laser thermotherapy-induced threshold coagulation of the ciliary body. AB - BACKGROUND AND OBJECTIVE: To study the effect of indocyanine green (ICG) pretreatment on threshold parameters of transscleral diode laser thermotherapy induced threshold coagulation of the ciliary body. The procedure was termed 'cyclothermotherapy' based on the long duration (15-60 seconds) of diode laser application. STUDY DESIGN/MATERIALS AND METHODS: The right eyes of nine young adult New Zealand white rabbits underwent transscleral cyclothermotherapy (TCT, Group 1), TCT following ICG pretreatment (Group 2), and external manipulation of the ciliary body alone (Group 3). Rabbits were sacrificed after 24 hours; specimens were evaluated with gross examination and light microscopy. RESULTS: Thresholds were 30 J/cm2 (TCT) and 4.5 J/cm2 (TCT with ICG). Widespread structural damage was seen in the ciliary processes and the ciliary body in Groups 1 and 2. In Group 3, external manipulation of the ciliary body caused hemorrhage and structural damage confined to the ciliary processes. CONCLUSION: ICG pretreatment reduced the energy necessary to cause a threshold lesion with TCT in nonpigmented rabbits. PMID- 15334622 TI - Acetylation phenotype status in a Bangladeshi population and its comparison with that of other Asian population data. AB - The objective of the present study was to determine the acetylator status of the Bangladeshi population and to compare the findings with the acetylator status of other Asian populations. The acetylator phenotype was determined in 517 unrelated healthy Bangladeshi subjects. The phenotyping procedure was done according to Price Evans' method using the NAT2 specific probe drug--sulphadimidine. The Bangladeshi population showed a bimodal distribution of fast and slow acetylators. Of a total of 517 healthy Bangladeshi, 79.5% (n=411) were fast acetylators and the rest 20.5% (n=106) were slow acetylators. The high frequency of the fast acetylators in the population of Bangladesh was comparable to other populations in East Asia. When this acetylator status was compared with other Asian data, the Asian population showed a positive correlation between the acetylator status and the geographical longitude (r=0.919; t=7.37; p>0.001; d.f.=10). The regression line of the scatter diagram showed that the frequency of acetylating capacity increasingly occurred in the populations towards eastern Asia (regression coefficient=0.54; constant=52.36). This line was termed as the Asian fast acetylator longitude (AFAL). Thus the AFAL was able to predict the acetylator status of the Asian population by the east-west geographical longitude. The AFAL could be a useful prognosticator in the disposition for the effective and safe use of numerous drugs and xenobiotic compounds in humans. PMID- 15334623 TI - Identification of human UDP-glucuronosyltransferase enzyme(s) responsible for the glucuronidation of 3-hydroxydesloratadine. AB - Desloratadine is a non-sedating antihistamine recently approved for the treatment of seasonal allergic rhinitis. The major metabolite of desloratadine in human plasma and urine is the glucuronide conjugate of 3-hydroxydesloratadine. 3 Hydroxydesloratadine-glucuronide is also the major in vitro metabolite of 3 hydroxydesloratadine formed by incubation of 3-hydroxydesloratadine with human liver microsomes supplemented with uridine 5'-diphosphate-glucuronic acid (UDPGA). The metabolite structure was confirmed by LC-MS and LC-MS/MS. Out of ten recombinant human UDP-glucuronosyltransferases (UGTs), UGT1A1, UGT1A3, UGT1A8 and UGT2B15 exhibited catalytic activity with respect to the formation of 3 hydroxydesloratadine-glucuronide. Inhibition studies with known inhibitors of UGT (diclofenac, flunitrazepam and bilirubin) confirmed the involvement of UGT1A1, UGT1A3 and UGT2B15 in the formation of 3-hydroxydesloratadine-glucuronide. The results from this study demonstrated that the in vitro formation of 3 hydroxydesloratadine-glucuronide from 3-hydroxydesloratadine was mediated via UGT1A1, UGT1A3 and UGT2B15 in human liver. PMID- 15334624 TI - Absorption, distribution, metabolism and excretion of YM466, a novel factor Xa inhibitor, in rats. AB - YM466 is a novel factor Xa inhibitor for the treatment of thrombosis. The absorption, distribution, metabolism and excretion of YM466 were investigated in male Fisher rats after a single oral administration. YM466 was absorbed rapidly from all segments of the gastrointestinal tract except the stomach. After oral dosing, the plasma concentration of (14)C-YM466 reached a maximum within 0.5 h, and declined rapidly with an elimination half-life of 0.64 h. The unchanged YM466 accounted for almost all of its radioactivity, suggesting a minimal metabolism in rats. This was also supported by the finding that no metabolites were observed in bile and urine after oral dosing of (14)C-YM466. The distribution of (14)C-YM466 in tissue was evaluated and the liver and kidney were the organs with radioactivity concentrations consistently higher than that of plasma. Cumulative biliary and urinary excretion of radioactivity in bile duct-cannulated rats was 29.5% and 7.6%, respectively, indicating prominent excretion into bile after oral dosing. This was consistent with the finding that 76.1% and 25.2% of radioactivity dosed were excreted to faeces and urine, respectively, after i.v. dosing. These results suggest that YM466 was rapidly absorbed and then subjected to biliary excretion with a minimal metabolism after oral dosing to rats. PMID- 15334625 TI - Pharmacokinetics and dose proportionality of extended-release metformin following administration of 1000, 1500, 2000 and 2500 mg in healthy volunteers. AB - The pharmacokinetics and dose-exposure relationship of an extended-release formulation of metformin (ER-metformin) was investigated in a randomized, single dose, four-period crossover study in 24 healthy male volunteers. During each study period, subjects received a randomly assigned dose containing 1000, 1500, 2000 or 2500 mg metformin. Blood samples were drawn 0-72 h after dosing for pharmacokinetic and dose-proportionality assessment. Although several pairwise comparisons between dose groups were significant (p<0.05) with respect to dose normalized C(max), AUC(0-72 h), and AUC( infinity ), the magnitude of the difference across the dose range was <20% for AUC(0-72 h) and AUC( infinity ), and was < or = 30% for C(max). The results indicate a consistent and predictable increase in metformin exposure with an extended-release formulation of metformin over 1000 to 2500 mg. PMID- 15334626 TI - The plasma pharmacokinetics and cerebral spinal fluid penetration of intravenous topiramate in newborn pigs. AB - INTRODUCTION: Pre-clinical studies suggest that the anticonvulsant topiramate confers neurologic protection against ischemia. An intravenous formulation of topiramate has been developed for administration during conditions such as hypoxia-ischemia when enteral absorption may be unpredictable. The plasma pharmacokinetics, cerebrospinal fluid (CSF) penetration and pharmacodynamics of intravenous topiramate were studied in an established piglet model of hypoxia ischemia. METHODS: The plasma pharmacokinetics of topiramate were studied in a group of chronically instrumented conscious piglets (n = 8), and in a group of piglets following an episode of hypoxia-ischemia (n = 8). These groups were divided into equal dose cohorts in which two doses of intravenously administered topiramate, 5 and 40 mg/kg, were studied. The animals' heart rate, arterial pressure and EEG were monitored. Plasma for topiramate concentration was sampled for up to 26 h. A single CSF topiramate concentration was determined 1 h following drug administration. Topiramate was quantified using a specific LC/MS assay. RESULTS: The animals tolerated intravenous topiramate well, with no significant changes in physiologic and neurologic parameters. Plasma topiramate concentrations following an intravenous dose were best described by a bi exponential equation, with a mean clearance of 39+/-18 ml/h/kg, and a terminal half-life of 14.3 (range 7.5-48.1) h. A dose of 5 mg/kg was sufficient to maintain plasma drug concentrations greater than 10 micro M for 24 h. CSF topiramate concentration at 1 h was 12+/-1 micro M and 109+/-26 micro M at the 5 and 40 mg/kg doses, respectively. CONCLUSION: Topiramate administered intravenously was well tolerated. Slow clearance of the drug allowed for maintenance of potential neuroprotective concentrations following a single dose of drug for 24 h. High drug penetration into the CSF is an ideal pharmacologic characteristic of any potential neuroprotective agent. The pharmacokinetic profile of intravenously administered topiramate, including its penetration into the CSF, appears to achieve this goal. PMID- 15334627 TI - Pharmacokinetics of clarithromycin in rats with acute renal failure induced by uranyl nitrate. AB - In rats pretreated with dexamethasone (an inducer of CYP3A1/2 in rats) and troleandomycin (an inhibitor of CYP3A1/2 in rats), the area under the plasma concentration-time curve from time zero to time infinity (AUC) values of clarithromycin were significantly smaller (365 compared with 600 micro g min/ml) and greater (1410 compared with 581 micro g min/ml), respectively, than those in control rats. This indicated that clarithromycin was metabolized via CYP3A1/2 in rats. The expression of CYP3A1(23) increased in rats with acute renal failure induced by uranyl nitrate (rats with U-ARF). Hence, it could be expected that AUC of clarithromycin could be smaller in rats with U-ARF. However, after intravenous administration of clarithromycin at a dose of 20mg/kg, the AUC and time-averaged total body (Cl) and nonrenal (Cl(nr)) clearance values were comparable between the two groups of rats. The 9000 x g supernatant fraction of liver homogenates in rats with U-ARF had comparable metabolic activities for clarithromycin compared with those in control rats, suggesting that the CYP3A isozyme responsible for metabolism of clarithromycin seemed not to be expressed considerably in the rats. This could explain the comparable AUC, Cl and Cl(nr) values of clarithromycin between the two groups of rats. PMID- 15334628 TI - Rationale for superficial injection techniques in lymphatic mapping in breast cancer patients. AB - One of the most avidly debated issues in lymphatic mapping is where the tracers are best deposited in patients with breast cancer. The four superficial approaches are easy to perform and have several other distinct advantages. They are based on the hypothesis that the entire breast parenchyma and the overlying skin drain to a common node in the axilla because of their common embryological origin. Evidence is presented that casts doubt upon the correctness of this assumption. Tracer administration close to the tumor site appears to be the safest approach for the time being. Excellent results can be obtained with this latter approach, despite the fact that it is technically more demanding. PMID- 15334629 TI - Survival of patients with untreated rectal cancer. AB - BACKGROUND AND OBJECTIVES: Untreated rectal cancer is uncommon given modern surgical techniques and chemoradiotherapy. However, neglected cancer may still occur in patients with limited access to medical care or who refuse conventional interventions. Outcome data on such patients is scarce, though valuable in educating patients on their prognosis or to compare with present therapeutic options. METHODS: To identify such data, a review was made of archival literature documenting outcomes in patients with untreated rectal cancer. A review of the Index Medicus and its predecessor volumes and an extensive MEDLINE search led to nine articles that contained relevant age at presentation data of patients with rectal cancer. Six articles also included patient survival data for untreated rectal cancer. Actuarial analysis of the data through 5 years post-diagnosis was completed. RESULTS: Symptoms most often presented in the fifth and sixth decades. Median survival of the 1326 patients fell within the second year after presentation of symptoms. Five-year actuarial survival for these patients was 4.4%. When the data published prior to 1925 were analyzed separately from that published post-1925, two survival patterns emerged. Median survival of pre-1925 patients fell within the second year after presentation of symptoms (at 21.5 months by curve fitting) and of post-1925 patients fell within the first year (10.2 months). CONCLUSIONS: Clearly, advances in antibiotics, surgery, chemoradiotherapy, and supportive care have contributed to increased survival for patients with treated rectal cancer. Recognizing the hazards of generalizing too extremely from the data, they reveal that relatively long survival for patients with untreated lesions is possible. PMID- 15334630 TI - Utility of CEA and CA 19-9 tumor markers in diagnosis and prognostic assessment of mucinous epithelial cancers of the appendix. AB - BACKGROUND AND OBJECTIVES: Tumor markers are a clinical tool frequently used in oncology in association with other clinical and radiologic information. For gastrointestinal cancer, carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9) tumor markers have found selected clinical application. The use of these tumor markers in mucinous epithelial tumors of the appendix has not been previously determined. METHODS: In patients with peritoneal dissemination of a mucinous epithelial malignancy of the appendix, tumor markers CEA and CA 19-9 were prospectively recorded preoperatively within 1 week prior to definitive treatment. Also, if the appendiceal tumor recurred, the tumor marker was determined. The accuracy of these two tumor markers in the management of this disease was determined for these two specific clinical situations. RESULTS: CEA was elevated in 56% of 532 patients and CA 19-9 was elevated in 67.1% of these patients. Although the absolute level of tumor marker did not correlate with prognosis, a normal value indicated an improved survival. CEA was elevated in 35.2% of 110 patients determined to have recurrent disease; CA 19-9 was elevated in 62.9% and at least one of the tumor markers was elevated in 68.2% of patients. An elevated CEA tumor marker at the time of recurrence indicated a reduced prognosis. CONCLUSIONS: Both CEA and CA 19-9 tumor markers were elevated in a majority of these patients and should be a valuable diagnostic tool previously underutilized in this group of patients. These tumor markers were also of benefit in the assessment of prognosis in that a normal level indicated an improved prognosis. At the time of a reoperative procedure, CEA and CA 19-9 tumor markers gave information regarding the progression of disease. These tumor markers have practical value in the management of epithelial appendiceal malignancy with peritoneal dissemination. PMID- 15334631 TI - Risk factors for peritoneal dissemination of colorectal cancer. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to clarify the risk factors associated with positive peritoneal dissemination (PPD) of colorectal cancer (CRC). METHODS: From June 2000 to September 2002, 143 CRC patients who underwent elective curative (79.0%) or non-curative (21.0%) open laparotomy were prospectively studied. Clinical evaluations including classical factors, colonoscopic evaluation, intraoperative evaluation, and pathological features were recorded. PPD was diagnosed when macro- (MAPD) or microscopic peritoneal dissemination (MIPD) was evident. Positive peritoneal cytology from initially existing ascites or washing lavage indicated MIPD. Various factors were analyzed with univariate (Chi-square test) and then multivariate analyses (logistic regression test) to search for the risk factors of PPD. RESULTS: Overall, MIPD, MAPD, and PPD were found in 2.8%, 6.3%, and 9.1%, respectively. Univariate analysis identified age (< or =59 years), CA19-9 (> or = 34.6 U/ml), poor differentiation, circumferential involvement (> or = 3 quadrants), ascites volume (>80 ml), pN+, and pT4 as risk factors of PPD. PPD did not occur in patients with well-differentiated tumors, less circumferential involvement (< 2 quadrants), or no lymph node metastasis. After multivariate analysis, CA19-9 (Odds ratio (95% CI), 8.6 (1.7-43.1)), pT4 (9.0 (1.3-61.0)), and age (5.26 (1.1-25.0)) remained significant risk factors. CONCLUSION: CA19-9 (> or = 34.6 U/ml), pT4, and age (< or =59 years) were significant risk factors of PPD. PMID- 15334632 TI - Heparanase expression in human colorectal cancer and its relationship to tumor angiogenesis, hematogenous metastasis, and prognosis. AB - BACKGROUND: Heparanase is considered to play an important role in tumor invasion and hematogenous metastasis. The aim of this study was to determine the expression of heparanase in colorectal cancer to evaluate its prognostic value. METHODS: We analyzed heparanase mRNA derived from 130 colorectal cancer patients by reverse transcription polymerase chain reaction (PCR), compared its expression with clinicopathologic factors, and performed statistical analysis. To investigate the relationship between heparanase expression and tumor angiogenesis, 81 primary colorectal tumors were immunostained by use of a monoclonal anti-CD34 antibody. RESULTS: Thirty three of 130 cancer tissues overexpressed heparanase. There were significant correlations between heparanase expression and serosal invasion, venous invasion, and liver metastasis. Multivariate analyzes revealed that heparanase mRNA overexpression was a significant independent risk factor for hematogenous metastasis in colorectal cancer. Among 104 patients who underwent curative resection, heparanase expression correlated with a high recurrence. The 5-year survival rate was 84.6% for patients with heparanase negative tumors, and 47.7% for those with heparanase overexpression; these differences between two groups of patients were significant. In multivariate analysis using the Cox regression model, heparanese expression emerged as an independent prognostic indicator. Moreover, the tumor angiogenesis of heparanase-positive tumors determined with a monoclonal anti-CD34 antibody was significantly higher than that of heparanase-negative tumors. CONCLUSIONS: These results indicated that Heparanase expression may be an important role in invasion and hematogenous metastasis, and may be a biologic marker of prognostic significance in colorectal cancer patients. PMID- 15334633 TI - Comparison of methylation-specific polymerase chain reaction (MSP) with reverse transcriptase-polymerase chain reaction (RT-PCR) in peripheral blood of gastric cancer patients. AB - BACKGROUND AND OBJECTIVES: To achieve a complete cure in gastric cancer, primary and recurrent tumors must be detected at an early stage. This study was designed to compare carcinoembryonic antigen (CEA)-specific reverse transcriptase polymerase chain reaction (RT-PCR) and methylation-specific polymerase chain reaction (MSP) for p16, E-cadherin, and retinoic acid receptor beta (RARbeta) genes using blood samples from gastric cancer patients. METHODS: Preoperative blood samples obtained from 41 patients with gastric cancer, including 9 with early-stage disease, and were subjected to RT-PCR and MSP assays. RESULTS: Ten of 41 (24%) patients exhibited a CEA-specific signal by RT-PCR. Positive rates were 11, 13, 50, and 50% in stages I, II, III, and IV, respectively. A significant association was found between RT-PCR results and stage (P< 0.01). The MSP assay detected hypermethylation of p16 in 9 patients (22%), E-cadherin in 9 patients (22%), and RARbeta in 6 patients (15%). Altogether, 18 patients (44%) showed hypermethylation. The positive rates were 37, 50, 40, and 75% in stages I, II, III, and IV, respectively. A significant association was found between aberrant methylation and venous invasion (P< 0.05). Neither the CEA-specific signal nor hypermethylation was detected in serum from control volunteers. CONCLUSIONS: The detection rate of MSP was higher than that of RT-PCR in gastric cancer. Both assays can serve as markers that allow selection of those cases requiring more intensive screening and aggressive postoperative treatment. PMID- 15334634 TI - FAP with concurrent duodenal adenomatous polyposis and carcinoid tumor. AB - We report a patient with familial adenomatous polyposis (FAP) who developed duodenal adenomatosis with high-grade dysplasia and a periampullary carcinoid tumor several years after total colectomy. Only two prior case reports exist revealing carcinoid tumors in association with FAP. No genetic basis exists explaining the link between FAP and carcinoid tumors. However, the presence of two rare entities in the same patient might suggest an association and further research may be indicated. PMID- 15334635 TI - Unknown primary. PMID- 15334636 TI - The world federation of surgical oncology societies: the global mission. PMID- 15334637 TI - Intra-abdominal metastases from soft tissue sarcoma. AB - OBJECTIVE: To define the clinical features and prognosis of patients with abdominal metastasis from primary soft tissue sarcoma (STS) at other sites. METHODS: All patients with abdominal metastasis from STS were identified from the Royal Marsden Hospital Sarcoma Unit prospective database from January 1990 to July 2001. RESULTS: Nineteen patients developed abdominal metastasis out of a cohort of 2127 patients (0.9%) evaluated during the study interval. The median age was 49 (19-71) years. The median time to abdominal metastasis from diagnosis of the primary was 27 (8-91) months. The presenting complaints were incomplete intestinal obstruction (n = 5), abdominal pain (n = 4), mass (n = 2), gastrointestinal bleed (n = 2), urinary obstruction (n = 2), anorexia (n = 1), and abdominal distension (n = 1). Emergency laparotomy was done for perforative peritonitis (n = 2), intussusception (n = 2), and bleed in spleen (n = 1). Two patients were asymptomatic. The common histologies were myxoid liposarcoma (n = 6) and leiomyosarcoma (n = 4). The median follow-up of survivors post metastasis was 12 months. Abdominal metastatectomy was performed in 16 patients, 3 of these patients had abdominal recurrences. The 1- and 2-year overall disease specific survival for the 19 patients was 66% (SE = 11%) and 43% (SE = 13%) with a median survival of 13 months (95% CI = 11.8-14.7). Metastasectomy was associated with slight improved median post-metastasis survival (33 months vs. 8 months for unresected patients). CONCLUSIONS: Although abdominal metastasis is rare, vigilance is warranted. Symptomatic patients should be examined and investigated thoroughly for metastases. Surgery is the treatment of choice for patients with an acute presentation; however, survival is dismal. PMID- 15334638 TI - Primary sarcoma of the breast. AB - BACKGROUND AND OBJECTIVES: Primary sarcoma occurring in breast is rare and comprises 0.5-1% of all breast neoplasm. Majority of the series include both stromal and cystosarcoma phyllodes, only a few hundred cases of sarcomas other then cystosarcoma are reported. PATIENTS AND METHODS: We carried out a retrospective analysis of 19 patients with primary sarcoma of the breast treated between 1982 and 2002. RESULTS: Mean age of the patients was 38.6 years (12-70 years). Gradually progressive swelling was the commonest presenting feature. There were eight cases of angiosarcoma, four cases of spindle cell sarcoma, two each of pleomorphic sarcoma and stromal sarcoma, and one each of malignant fibrous histiocytoma, embryonal rhabdomyosarcoma, and sarcoma (NOS). Eight of these were high-grade (42%). Eight patients underwent either radical or modified mastectomy, three underwent wide excisions, and one underwent quadrantectomy. Ten (52.6%) patients received postoperative adjuvant radiation. Two patients received chemotherapy. After a mean follow-up time of 34.5 months (median 25 months), eight patients failed. Failure was local in five, opposite breast in one, and both local and distant in two. The disease free survival at 3-year was 39%. In univariate analysis only the margin of first surgery was found to be a significant predictor of survival (P = 0.05). CONCLUSIONS: Primary sarcomas of the breast are aggressive tumors. Surgical treatment should consist of at least simple mastectomy. All attempts should be made to achieve a negative margin as this appears to be the only factor influencing survival in these patients. PMID- 15334639 TI - Clear cell sarcoma in the era of sentinel lymph node mapping. AB - Clear cell sarcoma of the tendons and aponeuroses (CCSTA) are rare aggressive soft tissue tumors with tendency for lymph nodes dissemination. Lymph node involvement is a correlate for prognosis. We present three patients with CCSTA in whom simultaneous sentinel lymph biopsy (SLNB) and resection was performed. Sentinel lymph node mapping may have a role in clear cell sarcoma from a prognostic standpoint. Further investigations are needed for validation. PMID- 15334640 TI - Identification of a single nucleotide polymorphism in the MMP-1 promoter in chondrosarcoma. AB - Matrix metalloproteinases (MMPs) are a group of at least 26 enzymes that facilitate cellular invasion via the degradation of the extracellular matrix. Specifically, the ability to degrade collagen types III and I is important in tumor invasion and metastasis. Over expression of the MMP-1 gene has been shown to correlate with poorer outcome in GI tract and gynecological tumors. This level of expression of this gene has been shown to be significantly increased by the presence of a single nucleotide polymorphism in the MMP-1 promoter sequence as a result of the creation of an ETS binding site. This SNP results from the addition of a single guanine base at -1,607 bp 24. Two chondrosarcoma cell lines and a series of 10 resected chondrosarcoma specimens underwent DNA extraction, purification, polymerase chain reaction, and sequencing. The presence of the single nucleotide polymorphism at -1,607 bp was confirmed within the promoter region for MMP-1 in human chondrosarcoma. Because all three genotypes were found in the clinical samples, the SNP may indeed provide a mechanistic explanation for a more aggressive biologic behavior locally and distally for a subset of chondrosarcomas. PMID- 15334641 TI - Clinical patterns and treatment outcome of elderly patients in clinical stage IB/II non-small cell lung cancer. AB - BACKGROUND AND OBJECTIVES: Surgery is a standard treatment in patients with clinical stage IB/II non-small cell lung cancer (NSCLC). We often have difficulty in treating of elderly patients due to their insufficient physiological function. To better manage such elderly patients, the clinical characteristics and prognosis of patients with these stages, who were 75 years of age or older, were reviewed. METHODS: From 1972 to 1999, 112 elderly patients with these stages were treated in our department. These patients comprised 88 men and 24 women. The histological types were 50 adenocarcinomas, 51 squamous cell carcinomas, 8 large cell carcinomas, and 3 adenosquamous carcinomas. RESULTS: Seventy-four patients (66%) underwent a surgical resection, including 60 surgery alone, 14 combined modality therapy. Radiotherapy, with or without chemotherapy, was given to 30 patients (27%), and chemotherapy alone to 5 (4.5%). In addition, 3 (2.7%) were given no therapy. The survivals of the surgery group at 2 and 5 years are 53% and 21% and those of the radiotherapy group are 35% and 3%, respectively. A multivariate analysis in radiotherapy group shows the predominant prognostic factor to be adenocarcinoma. The 2-year survival of the radiotherapy group in patients with adenocarcinoma is 58%, while that of patients with squamous cell carcinoma is 22%. CONCLUSIONS: These above observations suggest that radiotherapy is an alternative strategy for patients who cannot undergo surgery, especially with adenocarcinoma. PMID- 15334642 TI - Management of hepatocellular carcinoma in renal transplant recipients. AB - BACKGROUND AND OBJECTIVE: In Hong Kong where hepatitis B virus (HBV) infection is endemic, hepatocellular carcinoma (HCC) accounts for 20% of all malignant transformations in renal transplant recipients. The aim of the present study was to review the management and outcome of HCC in renal transplant recipients at a specialized surgical center. METHOD: A retrospective analysis on the data collected prospectively in a tertiary referral center. RESULTS: From January 1991 to December 2002, five renal transplant recipients were diagnosed to have primary HCC and received treatment in our center. There were four men and one woman with a median age of 47 (range, 38-68) years. Four of them had cadaveric renal transplantation whereas one had live donor transplantation. All of them were HBV carriers. The median tumor size was 3.5 cm (range, 1.8-8 cm). All tumors, except one, were diagnosed in sub-clinical stage by surveillance serum alpha-fetoprotein assay and percutaneous ultrasonography. Four patients were treated with surgical resection and one received transarterial oily chemoembolization (TOCE) as their primary treatments. There was one peri-operative death and the remaining three surgically treated patients were alive 4, 62, and 64 months after the resection. One patient developed recurrence 18 months after curative resection and was treated with TOCE. The patient with unresectable disease was alive for 50 months after the initial diagnosis. The surgical resection and overall survival rates of these patients were better than the published results. CONCLUSION: Early detection with regular serum alpha-fetoprotein assay and ultrasonographic study, vigilant care in the peri-operative period, long-term follow-up for detection and treatment of recurrence, as well as close collaboration between renal physicians and liver surgeons may improve the outcome of treatment of HCC in renal transplant recipients. PMID- 15334643 TI - Anti-Ri antibody opsoclonus-myoclonus syndrome and breast cancer: a case report and a review of the literature. AB - Opsoclonus-myoclonus is a rare neurological paraneoplastic syndrome associated with breast cancer and the presence of anti-Ri antibody. We presented a case of a 79-year-old woman with this syndrome and a small invasive ductal carcinoma [pT1aN0(sn)M0], whose symptoms improved 3 months following her simple mastectomy without any adjuvant therapy. Based on the 15 previously reported cases in the literature, there is no uniform treatment option available, and the response to such treatments is mixed. PMID- 15334644 TI - Kaposi's sarcoma: an update. AB - While there have been many important advances in the study of Kaposi's sarcoma (KS), it remains both a challenge and an enigma in many ways. Kaposi's original description of "multiple idiopathic hemorrhagic sarcoma[s]" in patients who died within 2-3 years resembles KS in AIDS more than classic KS in elderly men of Italian, Jewish, or Mediterranean lineage, in whom the disease is usually benign. KS had been evident in about one-third of those with early AIDS, often as its presenting sign, a pattern markedly reduced in recent times since the introduction of highly active anti-retroviral therapy (HAART). The most important advance has been the convincing etiologic linkage of KS with human herpesvirus 8 (HHV-8), which is necessary but not sufficient. It has a low prevalence in the general population of the USA and UK, with an intermediate rate in Italy and Greece, and a high one in Uganda. KS risk may be significantly lower in AIDS patients with a history of anti-herpes therapy. Many aspects of HHV-8, including its transmission pattern and different genospecies, are being scrutinized. The diagnosis of KS may be difficult. One should be aware of KS clinical variants, including telangiectatic, eccymotic, and keloidal KS. One must consider a number of other disorders, including bacillary angiomatosis. HHV-8 DNA sequences in dermatofibromas and other tumors should probably not be viewed as representing a marker for KS. Therapeutic options vary for KS. Intralesional and low-dose outpatient intravenous vinblastine may be valuable, as immunosuppression with KS is not a good idea if it can be avoided. Anti-herpes virus therapy may have potential for wide use, especially in preventing the development of KS in at risk populations, such as HHV-8 seropositive individuals undergoing transplantation surgery. PMID- 15334645 TI - Mouse alpha1B-adrenergic receptor is expressed in neurons and NG2 oligodendrocytes. AB - alpha1-Adrenergic receptors (ARs) are well-known mediators of the sympathetic nervous system, are highly abundant in the brain, but are the least understood in the central nervous system. The particular cell types in the brain that contain these receptors or their functions are not known because of the lack of high avidity antibodies and selective ligands. We developed transgenic mice that endogenously overexpress the alpha1B-AR subtype fused with the enhanced green fluorescent protein (EGFP). Endogenous expression was obtained by using a 3.4 kb fragment of the mouse alpha1B-AR promoter. Using this model, we determined cellular localization of the alpha1B-AR throughout the brain. The alpha1B-AR-EGFP fusion protein is expressed in neurons throughout the brain and in the Purkinje cells of the cerebellum. The alpha1B-AR is also expressed in NG2 oligodendrocyte precursor cells in both neonatal cell cultures and in the adult cerebral cortex, but is weakly expressed in mature oligodendrocytes. The alpha1B-AR was not observed in astrocytes or in cerebral vascular smooth muscle, cell types previously suggested to contain alpha1-ARs. We conclude that the alpha1B-AR is highly abundant throughout the brain, predominately in neurons, and may be involved in the development of the oligodendrocyte. In adult NG2 cells, implicated in stem cell-like functions, the alpha1B-AR may also play a role. This is the first report of a transgenic tagged-GPCR approach to determine in vivo localization of a receptor. PMID- 15334646 TI - Localization of neurokinin 1 receptor (NK1R) immunoreactivity in rat esophagus. AB - The aim of the present immunohistochemical study was to investigate the localization of neurokinin 1 receptor (NK1R) in rat esophagus and examine the relationship between NK1Rs and intrinsic cholinergic, nitrergic, or substance P (SP) neurons. NK1R immunoreactivity (IR) was observed on the nerve cell bodies in the myenteric ganglia throughout the esophagus, but not on striated muscles and smooth muscle cells of the muscularis mucosae. The frequency of occurrence of NK1R neurons was highest in the cervical esophagus and lowest in the lower thoracic esophagus. Considerable immunoreactivity was seen on the nerve cell surfaces and was also present in the cytoplasm of cell somas and in the initial part of the axons, but not in any other nerve fibers or terminals. Dogiel type I like morphology was observed in some of the NK1R neurons; however, the majority exhibited polymorphic morphology. Double immunolabeling indicated that a majority (77%) of the NK1R neurons were immunoreactive for choline acetyltransferase (ChAT), while a minority (23%) were immunoreactive for nitric oxide synthase (NOS)-IR. Most of the NK1R neurons (92%) were innervated by the SP nerve fibers. Triple immunolabeling indicated that 70% of the NK1R neurons were associated with intrinsic SP nerve fibers (without CGRP-IR), 59% were associated with extrinsic SP nerve fibers (with CGRP-IR), and 35% were associated with both intrinsic and extrinsic SP nerve fibers. These results suggest that SP/tachykinin released from the SP nerve fibers of intrinsic and/or extrinsic origin activates the predominantly intrinsic cholinergic neurons via NK1Rs to influence neuronal transmission or motility in rat esophagus. PMID- 15334647 TI - Hypothalamic paraventricular nucleus neurons regulate medullary catecholamine cell responses to restraint stress. AB - Both physical and psychological stressors recruit catecholamine cells (CA) located in the ventrolateral medulla (VLM) and the nucleus of the solitary tract (NTS). In the case of physical stressors, this effect is initiated by signals that first access the central nervous system at or below the level of the medulla. For psychological stressors, however, CA cell recruitment depends on higher structures within the neuraxis. Indeed, we have recently provided evidence of a pivotal role for the medial amygdala (MeA) in this regard, although such a role must involve a relay, as MeA neurons do not project directly to the medulla. However, some of the MeA neurons that respond to psychological stress have been found to project to the hypothalamic paraventricular nucleus (PVN), a structure that provides significant input to the medulla. To determine whether the PVN might regulate medullary CA cell responses to psychological stress, animals were prepared with unilateral injections of the neurotoxin ibotenic acid into the PVN (Experiment 1), or with unilateral injections of the retrograde tracer wheat germ agglutinin-gold (WGA-Au) into the CA cell columns of the VLM or NTS (Experiment 2). Seven days later, animals were subjected to a psychological stressor (restraint; 15 minutes), and their brains were subsequently processed for Fos plus appropriate cytoplasmic markers (Experiment 1), or Fos plus WGA-Au (Experiment 2). PVN lesions significantly suppressed the stress-related induction of Fos in both VLM and NTS CA cells, whereas tracer deposits in the VLM or NTS retrogradely labeled substantial numbers of PVN cells that were also Fos-positive after stress. Considered in concert with previous results, these data suggest that the activation of medullary CA cells in response to psychological stress may involve a critical input from the PVN. PMID- 15334648 TI - Morphology and axonal projection pattern of neurons in the telencephalon of the fire-bellied toad Bombina orientalis: an anterograde, retrograde, and intracellular biocytin labeling study. AB - The connectivity and cytoarchitecture of telencephalic centers except dorsal and medial pallium were studied in the fire-bellied toad Bombina orientalis by anterograde and retrograde biocytin labeling and intracellular biocytin injection (total of 148 intracellularly labeled neurons or neuron clusters). Our findings suggest the following telencephalic divisions: (1) a central amygdala-bed nucleus of the stria terminalis in the caudal midventral telencephalon, connected to visceral-autonomic centers; (2) a vomeronasal amygdala in the caudolateral ventral telencephalon receiving input from the accessory olfactory bulb and projecting mainly to the preoptic region/hypothalamus; (3) an olfactory amygdala in the caudal pole of the telencephalon lateral to the vomeronasal amygdala receiving input from the main olfactory bulb and projecting to the hypothalamus; (4) a medial amygdala receiving input from the anterior dorsal thalamus and projecting to the medial pallium, septum, and hypothalamus; (5) a ventromedial column formed by a nucleus accumbens and a ventral pallidum projecting to the central amygdala, hypothalamus, and posterior tubercle; (6) a lateral column constituting the dorsal striatum proper rostrally and the dorsal pallidum caudally, and a ventrolateral column constituting the ventral striatum. We conclude that the caudal mediolateral complex consisting of the extended central, vomeronasal, and olfactory amygdala of anurans represents the ancestral condition of the amygdaloid complex. During the evolution of the mammalian telencephalon this complex was shifted medially and involuted. The mammalian basolateral amygdala apparently is an evolutionary new structure, but the medial portion of the amygdalar complex of anurans reveals similarities in input and output with this structure and may serve similar functions. PMID- 15334649 TI - Expression of vanilloid receptor TRPV1 in the rat trigeminal sensory nuclei. AB - Little is known about the central projection patterns of trigeminal afferent neurons expressing the vanilloid receptor TRPV1 and their coexpression of neuromodulatory peptides. To address these issues, we examined the distribution of TRPV1-positive neurons in the trigeminal ganglion (TG) and trigeminal sensory nuclei principalis (Vp), oralis (Vo), interpolaris (Vi), and caudalis (Vc) in the rat via light and electron microscopy. In addition, we studied the colocalization of TRPV1-positive neurons with substance P (SP) and calcitonin gene-related peptide (CGRP) via confocal microscopy. In TG, only small and medium-sized neurons were immunopositive for TRPV1. The staining for TRPV1 was found in axon collaterals in the dorsal parts of Vp, Vo, and Vi and in terminals and fibers throughout lamina I and the outer zone of lamina II (IIo) of Vc. With electron microscopy, TRPV1-positive fibers in the ascending and descending trigeminal tracts were found to be unmyelinated. Almost all TRPV1-positive terminals in Vc contained numerous large dense-core vesicles and formed synaptic contacts with single small dendrites. Multiple immunofluorescence revealed a high degree of colocalization of TRPV1 with SP and CGRP in TG neurons as well as in fibers and terminals confined to laminae I and IIo of Vc. These results suggest that the central projections of unmyelinated (C) afferents sensitive to noxious heat and capsaicin are organized differently between Vc and the rostral trigeminal nuclei and that Vc may play a role in the development of hyperalgesia. PMID- 15334650 TI - Localization of myosin phosphatase target subunit 1 in rat brain and in primary cultures of neuronal cells. AB - Myosin phosphatase (PP1M) is composed of the delta isoform of the PP1 catalytic subunit (PP1cdelta), the myosin phosphatase target subunit (MYPT), and a 20 kDa subunit. Western blots detected higher amounts of the MYPT1 isoform compared to MYPT2 in whole brain extracts. The localization of MYPT1 was studied in rat brain and in primary cell cultures of neurons using specific antibodies. Analysis of lysates of brain regions for MYPT1 and PP1M by Western blots using anti-MYPT1 antibodies and by phosphatase assays with myosin as substrate suggested a ubiquitous distribution. Immunohistochemistry of tissue sections revealed that MYPT1 was distributed in all areas of the brain, with staining observed in many different cell types. Depending on the method used for fixation, the MYPT1 appeared with varying intensity in nuclei, in nucleoli, and in the cytoplasm. In primary hippocampal cultures, MYPT1 was identified by confocal microscopy in the cytoplasm and in the nucleus, whereas a predominantly cytoplasmic localization was found in cochlear nucleus cells. In cultured cells, MYPT1 and PP1cdelta colocalized with synaptophysin. PP1M activity was high in synaptosomes isolated from the cerebral cortex, but was relatively low in the postsynaptic densities. The interaction of MYPT1 with synaptophysin and with known partners (Rho-kinase, PP1cdelta) in brain extracts was shown by immunoprecipitation with anti-MYPT1. Pull-down assays from synaptosomes, using GST-MYPT1, also confirmed these interactions. In conclusion, the widespread cellular and subcellular localization of MYPT1 implies that PP1M may play an important role in the dephosphorylation of key regulatory proteins in neuronal cells. PMID- 15334651 TI - Neuroanatomical distribution of Huntingtin-associated protein 1-mRNA in the male mouse brain. AB - Huntingtin-associated protein 1 (HAP1) was identified as an interactor of the gene product (Huntingtin) responsible for Huntington's disease and found to be a core component of the stigmoid body. Even though HAP1 is highly expressed in the brain, detailed information on HAP1 distribution has not been fully described. Focusing on the neuroanatomical analysis of HAP1-mRNA expression using in situ hybridization histochemistry, the present study clarified its detailed regional distribution in the entire mouse brain. Mouse HAP1 (Hap1)-mRNAs were abundantly expressed in the limbic-related forebrain regions and midline/periventricular brainstem regions including the olfactory bulb, limbic-associated cortices, hippocampus, septum, amygdala, bed nucleus of the stria terminalis, preoptico hypothalamic regions, central gray, raphe nuclei, locus coeruleus, parabrachial nuclei, nucleus of the solitary tract, and area postrema. In contrast, little expression was detected in the striatum and thalamus, implying that Hap1 is associated with neurodegeneration-sparing regions rather than target lesions in Huntington's disease. The distribution pattern, resembling that of the stigmoid body, suggests that HAP1 and the stigmoid body are implicated in protection from neuronal death rather than induction of neurodegeneration in Huntington's disease, and that they play an important role in integrating instinct behaviors and underlying autonomic, visceral, arousal, drive, memory, and neuroendocrinergic functions, particularly during extensive homeostatic or emotional processes. These data will provide an important morphological base for a future understanding of functions of HAP1 and the stigmoid body in the brain. PMID- 15334652 TI - The role of substance P in inflammatory disease. AB - The diffuse neuroendocrine system consists of specialised endocrine cells and peptidergic nerves and is present in all organs of the body. Substance P (SP) is secreted by nerves and inflammatory cells such as macrophages, eosinophils, lymphocytes, and dendritic cells and acts by binding to the neurokinin-1 receptor (NK-1R). SP has proinflammatory effects in immune and epithelial cells and participates in inflammatory diseases of the respiratory, gastrointestinal, and musculoskeletal systems. Many substances induce neuropeptide release from sensory nerves in the lung, including allergen, histamine, prostaglandins, and leukotrienes. Patients with asthma are hyperresponsive to SP and NK-1R expression is increased in their bronchi. Neurogenic inflammation also participates in virus associated respiratory infection, non-productive cough, allergic rhinitis, and sarcoidosis. SP regulates smooth muscle contractility, epithelial ion transport, vascular permeability, and immune function in the gastrointestinal tract. Elevated levels of SP and upregulated NK-1R expression have been reported in the rectum and colon of patients with inflammatory bowel disease (IBD), and correlate with disease activity. Increased levels of SP are found in the synovial fluid and serum of patients with rheumatoid arthritis (RA) and NK-1R mRNA is upregulated in RA synoviocytes. Glucocorticoids may attenuate neurogenic inflammation by decreasing NK-1R expression in epithelial and inflammatory cells and increasing production of neutral endopeptidase (NEP), an enzyme that degrades SP. Preventing the proinflammatory effects of SP using tachykinin receptor antagonists may have therapeutic potential in inflammatory diseases such as asthma, sarcoidosis, chronic bronchitis, IBD, and RA. In this paper, we review the role that SP plays in inflammatory disease. PMID- 15334653 TI - IGF-I mRNA levels in bovine satellite cell cultures: effects of fusion and anabolic steroid treatment. AB - Androgenic and estrogenic steroids enhance muscle growth in a number of species; however, the mechanism by which anabolic steroids enhance muscle growth is not known. Castrated male cattle (steers) provide a particularly good model system in which to study the effects of anabolic steroids on muscle growth because they respond dramatically to treatment with both estrogens and androgens. The goal of this study was to determine if treatment of bovine satellite cell (BSC) cultures with 17beta-estradiol (E(2)) or trenbolone (a synthetic androgen) directly affects proliferation rate or level of mRNA for estrogen receptor (ER)-alpha, androgen receptor, and growth factors that have been shown to affect muscle growth (insulin-like growth factor (IGF)-I, IGF binding protein (IGFBP)-3, and myostatin). BSC cultures were established from the semimembranosus muscles of steers and then treated for 48 h with various concentrations of E(2) or trenbolone ranging from 0.001 to 10 nM. IGF-I mRNA levels in proliferating BSC cultures were significantly increased at 0.01 (1.9-times control values, P < 0.02) and at 0.1, 1, and 10 nM E(2) (2.9-, 3.5-, and 3.5-times control values, respectively, P < 0.0001). Additionally both 1 and 10 nM trenbolone increased IGF I mRNA levels to 1.7-times control values (P < 0.02). ER-alpha mRNA was detectable in BSC cultures, and levels were increased (2.3-times control levels, P < 0.001) in cultures treated with 0.001 nM E(2) but not in cultures treated with higher concentrations of E(2). Androgen receptor mRNA levels also were increased (1.5-times control levels, P < 0.02) in cultures treated with 0.001 nM trenbolone but not by treatment with higher concentrations of trenbolone. Levels of IGFBP-3 were increased (1.4-times control values, P < 0.02) by treatment with 0.001 nM E(2) but not by treatment with high concentrations of E(2). Myostatin mRNA levels were not affected by any concentration of either of the steroids. Although, levels of IGF-I mRNA were 10-times greater (P < 0.02) in fused BSC cultures than in proliferating cultures, treatment of fused cultures for 48 h with 10 nM E(2) increased IGF-I mRNA levels (2.5-times control levels, P < 0.02). Both E(2) and trenbolone increased (3)H-thymidine incorporation rate (1.5-times control levels, P < 0.001) in BSC cultures in media containing serum from which IGFBP-3 had been removed by anti-IGFBP-3 affinity chromatography. In summary, treatment of BSC cultures with either E(2) or trenbolone increased IGF-I mRNA level and proliferation rate, thus, establishing that these steroids have direct anabolic effects on cells present in the BSC culture. PMID- 15334654 TI - Plasmalemmal vacuolar H+-ATPase is decreased in microvascular endothelial cells from a diabetic model. AB - Angiogenesis requires invasion of extracellular matrix (ECM) proteins by endothelial cells and occurs in hypoxic and acidic environments that are not conducive for cell growth and survival. We hypothesize that angiogenic cells must exhibit a unique system to regulate their cytosolic pH in order to cope with these harsh conditions. The plasmalemmal vacuolar type H(+)-ATPase (pmV-ATPase) is used by cells exhibiting an invasive phenotype. Because angiogenesis is impaired in diabetes, we hypothesized that pmV-ATPase is decreased in microvascular endothelial cells from diabetic rats. The in vitro angiogenesis assays demonstrated that endothelial cells were unable to form capillary-like structures in diabetes. The proton fluxes were slower in cells from diabetic than normal model, regardless of the presence or absence of Na(+) and HCO(3) (-) and were suppressed by V-H(+)-ATPase inhibitors. Immunocytochemical data revealed that pmV-ATPases were inconspicuous at the plasma membrane of cells from diabetic whereas in normal cells were prominent. The pmV-ATPase activity was lower in cells from diabetic than normal models. Inhibition of V-H(+)-ATPase suppresses invasion/migration of normal cells, but have minor effects in cells from diabetic models. These novel observations suggest that the angiogenic abnormalities in diabetes involve a decrease in pmV-ATPase in microvascular endothelial cells. PMID- 15334655 TI - Role of endoplasmic reticulum calcium content in prostate cancer cell growth regulation by IGF and TNFalpha. AB - Variations in calcium concentration within the endoplasmic reticulum ([Ca(2+)](ER)) may play a role in cell growth. This study evaluates the regulation of calcium pools by growth modulators of prostate cancer (PC) cells, the insulin growth factor (IGF), and the tumor necrosis growth factor-alpha (TNFalpha) as well as evaluating the possible role of [Ca(2+)](ER) variations as signals for growth modulation. We show that IGF (5 ng/ml), which increases cell growth, induces an increase in [Ca(2+)](ER) whereas TNFalpha (1 ng/ml) which reduces cell proliferation and induces apoptosis, reduces [Ca(2+)](ER). IGF induced [Ca(2+)](ER) increase is correlated to an overexpression of the sarcoendoplasmic calcium-ATPase 2B (SERCA2b), whereas TNFalpha-induced [Ca(2+)](ER) decrease is associated to a reduction in SERCA2b expression. Pretreatment with epidermal growth factors (EGF) or IGF does not prevent TNFalpha from affecting the induction of apoptosis, [Ca(2+)](ER) reduction and SERCA2b downregulation. Reduction in [Ca(2+)](ER) induced by thapsigargin (TG) (from 1 pM to 1 microM, 48 h) reduces LNCaP growth in a dose dependent manner and induces apoptosis when cells are treated with 1 microM TG. We also show that a transient TG application (1 pM, 1 nM, 1 microM 15 min) is insufficient to induce a long lasting decrease in [Ca(2+)](ER), since [Ca(2+)](ER) remains identical to the control for 48 h following TG application. These treatments (1 pM and 1 nM, 15 min) do not modify cell growth. However, TG (1 microM, 15 min) induces apoptosis. We thus identify [Ca(2+)](ER) and SERCA2b as a central targets for causing LNCaP PC cell life or death induced by growth modulators. Furthermore our results indicate that calcium pool contents can regulate cell growth. PMID- 15334656 TI - Leptin inhibits lysophosphatidic acid-induced intracellular calcium rise by a protein kinase C-dependent mechanism. AB - Leptin communicates the status of body energy stores to the central nervous system, regulating appetite, metabolic rate, and neuroendocrine functions. These effects are mediated by leptin binding and activation of the cognate cell surface receptor, a member of type I cytokine receptor family, which lead to the activation of receptor-associated kinases of the Janus family. In this work, we demonstrate that leptin inhibits the l-alpha-lysophosphatidic acid (LPA)-induced intracellular calcium mobilization in a dose-dependent manner in HEK-293 cells stably expressing full-length leptin receptor (OB-Rb). This action appears to be selective, as it was not observed when other signaling families, such as VIP or EGF, were studied. Pretreatment with the phosphatidylinositol 3-kinase (PI3K) inhibitor, wortmannin, reversed the effect of leptin, pointing to PI3K as an intermediate molecule involved in this process. An unspecific protein kinase C (PKC) inhibitor, staurosporine, disrupted the inhibitory action of leptin. Furthermore, intracellular levels of phosphorylated PKCepsilon and PKCdelta rose to a maximum 5 min after leptin administration, suggesting that these atypical PKC isoforms are involved in the observed cross-desensitization. To define the regions of the OB-Rb intracellular domain required for the cross-desensitization, a series of C-terminal deletion mutants were transfected into HEK-293 cells. C terminal truncation that removed the consensus Box 3 motif of OB-Rb prevented leptin action, indicating that heterologous desensitization over LPA was exerted at the level of this intracellular motif. Our date demonstrate that leptin plays a key role in the regulation of the earliest signaling pathways activated by growth factors, such as LPA, through a signaling pathway involving PKCdelta and PKCepsilon coupled to Box 3 motif of the OB-Rb through PI3K. PMID- 15334657 TI - TRPC5 activation kinetics are modulated by the scaffolding protein ezrin/radixin/moesin-binding phosphoprotein-50 (EBP50). AB - TRPC1-7 proteins are members of a family of mammalian non-specific cation channels that mediate receptor-operated, phospholipase Cbeta/Cgamma dependent Ca(2+) influx in various cell types. TRPC4 and TRPC5 form a subfamily within TRPCs. Uniquely in the TRPC family, these channels possess a C-terminal "VTTRL" motif that binds to PDZ-domains of the scaffolding protein, EBP50 (NHERF1; Tang et al., J Biol Chem 275:37559-37564). The functional effects of EBP50 on TRPC4/5 activity have not been investigated. We have cloned rat TRPC5 (rTRPC5), functionally expressed it in HEK293 cell, and studied channel regulation with patch-clamp techniques. Both rTRPC5 and its VTTRL deletion mutant (r5dV) were localized to the plasma membrane. rTRPC5 did not display any significant basal activity in unstimulated HEK293 cells. In cells co-expressing rTRPC5 and H1 histamine receptor, rTRPC5 current evoked by GTPgammaS or histamine developed in two phases: a slowly developing, small inward current was followed by a rapidly developing, transient, large inward current. Each phase had a characteristic non linear current-voltage (I-V) relationship. Deletion of the VTTRL motif had no detectable effect on the biophysical properties of the channel. Co-expression of EBP50 with rTRPC5 caused a significant delay in the time-to-peak of the histamine evoked, transient large inward current. EBP50 did not modify the activation kinetics of the VTTRL-deletion mutant. We conclude that the VTTRL motif is not necessary for activation of TRPC5, but may mediate the modulatory effect of EBP50 on TRPC5 activation kinetics. PMID- 15334658 TI - Hepatocyte growth factor/scatter factor stimulates migration of muscle precursors in developing mouse tongue. AB - Hepatocyte growth factor (HGF) stimulates the migration of myogenic cells during the development of skeletal muscles. The inactivation of HGF genes or that of its receptor, c-met, in mice causes hypoplasia of skeletal muscle organs, such as the tongue. Basic fibroblast growth factor (FGF-2) also induces migration of skeletal myoblasts. A comparison of the functions of HGF and FGF-2 in myogenesis revealed the crucial effect of HGF in the development of skeletal muscles. Unlike FGF-2, HGF induced migration of myoblasts from the developing mouse tongue. The differences between the activities of HGF and FGF-2 were determined by comparing their effects on the expression of matrix metalloproteinase-9 (MMP-9) in myoblasts, C2C12 cells, cultured in collagen-coated dishes. The results showed that HGF, but not FGF-2, stimulated MMP-9 expression, and that the stimulation was mediated through the activation of phosphoinositide 3-kinase (PI3K) which was not associated with FGF-2 signal transduction. Nevertheless, both growth factors exerted almost the same effect on the reduction of myogenin expression in, and on the proliferation of, C2C12 cells, suggesting that HGF, rather than FGF-2, plays a crucial role in the generation of skeletal muscles, including the tongue. Moreover, the specific role of HGF through the PI3K signal pathway is the induction of MMP-9 expression in, and the migration of, myoblasts. PMID- 15334660 TI - Thyroid hormones regulate DNA-synthesis and cell-cycle proteins by activation of PKCalpha and p42/44 MAPK in chick embryo hepatocytes. AB - The molecular mechanism by which thyroid hormones exert their effects on cell growth is still unknown. In this study, we used chick embryo hepatocytes at different stages of development as a model to investigate the effect of the two thyroid hormones, T3 and T4, and of their metabolite T2, on the control of cell proliferation. We observed that T2 provokes increase of DNA-synthesis as well as T3 and T4, independently of developmental stage. We found that this stimulatory effect on the S phase is reverted by specific inhibitors of protein kinase C (PKC) and p42/44 mitogen-activated protein kinase (p42/44 MAPK), Ro 31-8220 or PD 98059. Furthermore, the treatment with thyroid hormones induces the activation of PKCalpha and p42/44 MAPK, suggesting their role as possible downstream mediators of cell response mediated by thyroid hormones. The increase of DNA-synthesis is well correlated with the increased levels of cyclin D1 and cdk4 that control the G1 phase, and also with the activities of cell-cycle proteins involved in the G1 to S phase progression, such as cyclin E/A-cdk2 complexes. Interestingly, the activity of cyclin-cdk2 complexes is strongly repressed in the presence of PKC and p42/44 MAPK inhibitors. In conclusion, we demonstrated that the thyroid hormones could modulate different signaling pathways that are able to control cell-cycle progression, mainly during G1/S transition. PMID- 15334659 TI - Dominant negative Ras enhances lactogenic hormone-induced differentiation by blocking activation of the Raf-Mek-Erk signal transduction pathway. AB - Epidermal growth factor (EGF) and Ras mitogenic signal transduction pathways are frequently activated in breast carcinoma and inhibit mammary differentiation and apoptosis. HC11 mouse mammary epithelial cells, which differentiate and synthesize beta-casein following growth to confluency and stimulation with lactogenic hormones, were used to study EGF-dependent signaling during differentiation. Blocking Mek-Erk or phosphotidylinositol-3-kinase (PI-3 kinase) signaling with specific chemical inhibitors enhanced beta-casein promotor-driven luciferase activity. Because EGF stimulation of HC11 cells resulted in the activation of Ras, the effect of activated Ras (RasV12) or dominant negative (DNRasN17) on lactogen induced differentiation was examined. HC11 cell lines expressing RasV12 or DNRasN17 under the control of a tetracycline (tet) responsive promotor were constructed. Activated RasV12 expression resulted in reduced tyrosine phosphorylation of Stat5 and a delay in beta-casein expression in response to prolactin. However, the expression of tet-regulated DNRasN17 and adenovirus-encoded DNRasN17 enhanced Stat5 tyrosine phosphorylation, Stat5 DNA binding, and beta-casein transcription. The expression of DNRasN17 blocked the activation of the Mek-Erk pathway by EGF but did not prevent the phosphorylation of AKT, a measure of activation of the PI-3-kinase pathway. Moreover, the expression of DNRasN17 prevented the block to lactogenic differentiation induced by EGF. Stimulation of HC11 cells with prolactin resulted in the association of the SHP2 phosphatase with Stat5, and this association was prevented by DNRasN17 expression. These results demonstrate that in HC11 cells DNRas inhibits the Mek Erk pathway and enhances lactogenic hormone-induced differentiation. This occurs, in part, by inhibiting the association of the SHP2 phosphatase with Stat5. PMID- 15334661 TI - Epithelial mesenchymal transition is a characteristic of hyperplasias and tumors in mammary gland from MMTV-Cripto-1 transgenic mice. AB - Epithelial-mesenchymal transition (EMT) facilitates migration and invasion of epithelial tumor cells. Cripto-1 (CR-1), a member of the epidermal growth factor CFC protein family increases migration of cells in vitro. Here the expression of molecular markers and signaling molecules characteristic of EMT were assessed in mammary gland hyperplasias and tumors from mice expressing the human CR-1 transgene by the MMTV promoter (MMTV-CR-1) and in mouse mammary epithelial cell line HC-11 overexpressing CR-1 (HC-11/CR-1). Western blot analysis showed decreased expression of E-cadherin in MMTV-CR-1 tumors and in HC-11/CR-1 cells. The expression of N-cadherin, vimentin, cyclin-D1, and of the zinc-finger transcription factor, snail, was increased in MMTV-CR-1 tumors. Increased snail mRNA was also found in HC-11/CR-1 cells. Expression of phosphorylated (P)-c-Src, P-focal adhesion kinase (FAK), P-Akt, P-glycogen synthease kinase 3beta (GSK 3beta), dephosphorylated (DP)-beta-catenin, and various integrins such as, alpha 3, alpha v, beta 1, beta 3, and beta 4 was also increased in MMTV-CR-1 tumors. Immunohistochemistry showed positive staining for vimentin, N-cadherin, cyclin D1, smooth muscle actin, fibronectin, snail, and beta-catenin in MMTV-CR-1 tumor sections. HC-11/CR-1 cells treated with the c-Src inhibitor PP2 reduced the expression of P-c-Src and of P-FAK, P-Akt, P-GSK-3beta, DP-beta-catenin all known to be activated by c-Src. Migration of HC-11/CR-1 cells was also reduced by PP2 treatment. These results suggest that CR-1 may play a significant role in promoting the increased expression of markers and signaling molecules associated with EMT. PMID- 15334662 TI - 8-Cl-cAMP and its metabolite, 8-Cl-adenosine induce growth inhibition in mouse fibroblast DT cells through the same pathways: protein kinase C activation and cyclin B down-regulation. AB - 8-Chloro-cyclic AMP (8-Cl-cAMP) is known to be most effective in inducing growth inhibition and differentiation of a number of cancer cells. Also, its cellular metabolite, 8-Cl-adenosine was shown to induce growth inhibition in a variety of cell lines. However, the signaling mechanism that governs the effects of 8-Cl cAMP and/or 8-Cl-adenosine is still uncertain and it is not even sure which of the two is the key molecule that induces growth inhibition. In this study using mouse fibroblast DT cells, it was found that adenosine kinase inhibitor and adenosine deaminase could reverse cellular growth inhibition induced by 8-Cl-cAMP and 8-Cl-adenosine. And 8-Cl-cAMP could not induce growth inhibition in the presence of phosphodiesterase (PDE) inhibitor, but 8-Cl-adenosine could. We also found that protein kinase C (PKC) inhibitor could restore this growth inhibition, and both the 8-Cl-cAMP and 8-Cl-adenosine could activate the enzymatic activity of PKC. Besides, after 8-Cl-cAMP and 8-Cl-adenosine treatment, cyclin B was down regulated and a CDK inhibitor, p27 was up-regulated in a time-dependent manner. These results suggest that it is not 8-Cl-cAMP but 8-Cl-adenosine which induces growth inhibition, and 8-Cl-cAMP must be metabolized to exert this effect. Furthermore, there might exist signaling cascade such as PKC activation and cyclin B down-regulation after 8-Cl-cAMP and 8-Cl-adenosine treatment. PMID- 15334663 TI - Characterization of the biological activities of uridine diphosphate in human dendritic cells: Influence on chemotaxis and CXCL8 release. AB - Uridine nucleotides are endogenous nucleotides which are released into the extracellular space from mechanical stressed endothelial and epithelial cells as well as lipopolysaccharide (lps)-stimulated monocytes. Here, we studied the biological activity of the selective purinoreceptor P2Y6 (P2YR6) agonist Uridine 5'diphosphate (UDP) as well as the P2YR2- and P2YR4-activating uridine 5'triphosphate (UTP) on human dendritic cells (DC). These cells in their immature state have the ability to migrate from blood to peripheral target sites where they sense dangerous signals and capture potential antigens. Moreover, mature DC induce innate immune responses and migrate from peripheral tissues to secondary lymphoid organs in order to activate naive T cells and initiate adaptive immunity. Here, we were able to show that uridine nucleotides stimulated Ca(2+) transients, actin polymerization, and chemotaxis in immature DC. Experiments with pertussis toxin, the stable pyrimidine agonist uridine 5'-O-(2-thiodiphosphate) (UDPgammaS) and receptor antagonists, as well as desensitization studies suggested that these uridine nucleotides activities were mediated by different G(i) protein-coupled receptors. During lps-induced maturation, DC lost their ability to respond towards uridine nucleotides with these activities. Instead, UDP, but not UTP, stimulated the release of the CXC-chemokine 8 (CXCL8) from mature DC in a reactive blue sensitive manner. Moreover, our study indicates that UDP stimulates different signaling pathways in immature and mature DC in order to favor the accumulation of immature DC and to augment the capacity to secrete CXCL8 in mature DC. PMID- 15334664 TI - Uncoupling protein-3 sensitizes cells to mitochondrial-dependent stimulus of apoptosis. AB - The mitochondrial uncoupling protein-3 is a member of the mitochondrial carrier protein family. As a homologue of the thermogenic brown fat uncoupling protein-1, it possesses a mitochondrial uncoupling activity and thus can influence cell energy metabolism but its exact biological function remains unclear. In the present study, uncoupling protein-3 was expressed in 293 cells using the tetracycline-inducible system and its impact on cell bioenergetics and responsiveness to the apoptotic stimulus was determined. The induction of uncoupling protein-3 expression in mitochondria did not lead to uncontrolled respiratory uncoupling in intact cells. However, it caused a GDP-inhibition of state 4 respiration and a GDP-induced re-polarization of the inner mitochondrial membrane in the presence of fatty acids, in agreement with its expected physiological behavior as an uncoupling protein (UCP). Uncoupling protein-3 expression did not cause apoptosis per se but increased the responsiveness of the cells to a mitochondrial apoptotic stimulus (i.e., addition of staurosporine in the culture medium). It enhanced caspase 3 and caspase 9 activation and favored cytochrome c release. Moreover, cells in which uncoupling protein-3 expression had been induced showed a higher mitochondrial Bax/Bcl-2 ratio essentially due to enhanced translocation of Bax from cytosol to mitochondria. Finally, the induction of uncoupling protein-3 also increased the sensitivity of mitochondria to open the permeability transition pore in response to calcium. It is concluded that the presence of uncoupling protein-3 in mitochondria sensitizes cells to apoptotic stimuli involving mitochondrial pathways. PMID- 15334665 TI - A 32 kDa protein--whose phosphorylation correlates with oncogenic Ras-induced cell cycle arrest in activated Xenopus egg extracts--is identified as ribosomal protein S6. AB - Oncogenic Ras induces cell-cycle arrest in mammalian cells and in fertilized Xenopus eggs. How oncogenic Ras induces cell-cycle arrest remains unclear. We previously showed that oncogenic Ras induces cell-cycle arrest in activated Xenopus egg extracts (cycling extracts) and that the induced cell-cycle arrest correlates with hyperphosphorylation of a 32 kDa protein. However, the identity of the 32 kDa protein was not known. By using a sucrose density-gradient centrifugation, Triton X-100-acetic acid-urea (TAU)-gel electrophoresis, composite agarose-polyacrylamide gel electrophoresis (CAPAGE), SDS-PAGE, and partial tryptic peptide sequence analysis, the 32 kDa protein has now been identified as S6, a 40S subunit ribosomal protein. Hence, our results indicate that the oncogenic Ras-induced cell-cycle arrest is correlated with hyperphosphorylation of S6, suggesting that phosphorylation of S6 plays an important role in the induced cell-cycle arrest. It has been shown that conditional deletion of gene encoding S6 in mammalian cells prevents proliferation, demonstrating the importance of S6 in cell proliferation. The exact role S6 plays in cell proliferation is unclear. However, phosphorylation of S6 has been implicated in the regulation of protein synthesis. Thus, our results are consistent with the concept that oncogenic Ras induces S6 phosphorylation to influence protein synthesis, thereby contributing to the cell-cycle arrest. In addition, our results also demonstrate that composite agarose-polyacrylamide gel electrophoresis is suitable for the separation of large molecular complexes. PMID- 15334666 TI - Cadmium uptake in rat hepatocytes in relation to speciation and to complexation with metallothionein and albumin. AB - Cadmium (Cd) uptake has been studied in primary cultures of rat hepatocytes focusing on the impact of inorganic and organic speciation. Uptake time-course studies over a 60-min exposure to 0.3 microM (109)Cd revealed a zero-time uptake and a slower process of accumulation which proceeds within minutes. (109)Cd uptake showed saturation kinetics (K(m) = 3.5 +/- 0.8 microM), and was highly sensitive to inhibition by Zn and Hg. There was no evidence for sensitivity to the external pH nor for any preferential transport of the free cation Cd(2+) over CdCl(n) (2-n) chloro-complexes. According to the assumption that only inorganic metal species are available, metal uptake decreased upon albumin (BSA) addition to the exposure media. In contrast, higher levels of (109)Cd accumulation were obtained under optimal conditions for Cd complexation by MT. Comparison among uptake data obtained under inorganic and organic conditions revealed that Cd-MT would be taken up 0.4 times as rapidly as Cd(inorg). We conclude that uptake of Cd in rat hepatocytes involves specific transport mechanism(s) subjected to Zn or Hg interactions. Uptake of inorganic Cd is not proportional to the levels of free Cd(2+) and does not involve the divalent cation transporter DCT1 nor the co transporter Fe(2+)-H(+) NRAMP2. We found Cd-MT but not Cd-BSA to be available for the liver cells, and have estimated a binding affinity four orders of magnitude higher for Cd complexation with MT compared to BSA; MT may have a significant role in Cd delivery to the liver. PMID- 15334667 TI - Why a World Journal of Gastroenterology? PMID- 15334668 TI - p53 polymorphism in human papillomavirus-associated Kazakh's esophageal cancer in Xinjiang, China. AB - AIM: To investigate the relationship between p53 codon 72 polymorphism and human papillomavirus (HPV) type 16 infection in Kazakh's esophageal cancer (EC) in Xinjiang, China. METHODS: Encoding regions of p53 codon 72 and HPV-16 E6 were amplified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction (PCR) methods using pairs of primary esophageal squamous cell carcinoma (SCC) tissue and corresponding normal mucosa, which were collected from 104 patients of Kazakh in Xinjiang, China. RESULTS: Only arginine allele was detected in 70.1% (39/55) of HPV-16-E6- positive cases but only in 40.8% (20/49) of HPV-16-E6-negative cases (P<0.05; OR, 3.53; 95% CI, 1.57-7.98). In contrast, such a significant correlation between p53 polymorphism and HPV infection was not evident in corresponding normal mucosae. The allele frequency of Arg allele in cancer cases (0.68) was higher than that in normal mucosa samples (0.54) (P<0.05; OR, 1.80; 95% CI, 1.21-2.69). CONCLUSION: p53 codon 72 Arg homozygous genotype is one of the high-risk genetic factors for HPV associated SCC of Kazakh. Individuals carrying Arg allele compared to those with Pro allele have an increased risk for esophageal SCC. PMID- 15334669 TI - Inhibitory effect of ubiquitin-proteasome pathway on proliferation of esophageal carcinoma cells. AB - AIM: To investigate the inhibitory effect of ubiquitin-proteasome pathway (UPP) on proliferation of esophageal carcinoma cells. METHODS: Esophageal carcinoma cell strain EC9706 was treated with MG-132 to inhibit its UPP specificity. Cell growth suppression was evaluated with 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. DNA synthesis was evaluated by (3)H-thymidine ((3)H-TdR) incorporation. Morphologic changes of cells were observed under microscope. Activity of telomerase was examined by telomeric repeat amplification protocol (TRAP) of PCR-ELISA. Cell cycle and apoptosis were detected by flow cytometry (FCM). DNA fragment analysis was used to confirm the presence of apoptosis. Expression of p27(kip1) was detected by immunocytochemical technique. RESULTS: After exposed to MG-132, the growth and value of (3)H-TdR incorporation of EC9706 cells were obviously inhibited. Cells became round, small and exfoliative under microscope. TRAP PCR-ELISA showed that light absorption of cells gradually decreased after exposed to 5 micromol/L of MG-132 for 24, 48, 72 and 96 h (P<0.01). The percentage of cells at G(0)/G(1) phase was increased and that at S and G(2)/M was decreased (P<0.01). The rate of apoptotic cells treated with 5 micromol/L of MG-132 for 48 and 96 h was 31.7% and 66.4%, respectively. Agarose electrophoresis showed marked ladders. In addition, the positive signals of p27(kip1) were located in cytoplasm and nuclei in MG-132 group in contrast to cytoplasm staining in control group. CONCLUSION: MG-132 can obviously inhibit proliferation of EC9706 cells and induce apoptosis. The mechanisms include upregulation of p27(kip1) expression, G(1) arrest and depression of telomerase activity. The results indicate that inhibiting UPP is a novel strategy for esophageal carcinoma therapy. PMID- 15334670 TI - Are gap junction gene connexins 26, 32 and 43 of prognostic values in hepatocellular carcinoma? A prospective study. AB - AIM: To investigate the prognostic value of the expression of connexin (Cx) 26, 32 and 43 messenger RNA (mRNA) in hepatocellular carcinoma (HCC) tissues. METHODS: Using a reverse-transcriptase polymerase chain reaction (RT-PCR), Cx 26, Cx 32 and Cx 43 mRNAs were determined in the liver tissues of 15 controls and in HCC tissues of 25 patients undergoing curative hepatic resection. The patients were followed up clinically. RESULTS: Cx 26 and Cx 32 mRNAs were significantly lower in HCC tissues compared with controls (both P<0.01). By multivariate analysis, a lower level of Cx 26 and Cx 32 mRNA correlated significantly with a risk of HCC recurrence (P = 0.033) and recurrence-related mortality (P = 0.031, P = 0.031). Cx 43 mRNA was higher in HCC tissues compared with controls but did not correlate with postoperative recurrence or recurrence-related mortality. Other significant predictors of HCC recurrence included cellular dedifferentiation (P = 0.033), less encapsulation (P = 0.050), vascular permeation (P = 0.046), and daughter nodules (P = 0.046). Significant variables related to recurrence-related mortality consisted of cell dedifferentiation (P = 0.031), vascular permeation (P = 0.048), and daughter nodules (P = 0.048). The levels of Cx 26 and Cx 32 mRNAs correlated significantly with cell differentiation (P = 0.031). CONCLUSION: A low expression of Cx 26 and Cx 32 mRNAs in HCC tissues is predictive of postoperative recurrence of HCCs. PMID- 15334671 TI - Postoperative adjuvant arterial chemoembolization improves survival of hepatocellular carcinoma patients with risk factors for residual tumor: a retrospective control study. AB - AIM: To evaluate the effect of postoperative adjuvant transcatheter arterial chemoembolization (TACE) on the prognosis of hepatocellular carcinoma (HCC) patients with or without risk factors for the residual tumor. METHODS: From January 1995 to December 1998, 549 consecutive HCC patients undergoing surgical resection were included in this research. There were 185 patients who underwent surgical resection with adjuvant TACE and 364 patients who underwent surgical resection only. Tumors with a diameter more than 5 cm, multiple nodules, and vascular invasion were defined as risk factors for residual tumor and used for patient stratification. Kaplan-Meier method was used to analyze survival curve and Cox proportional hazard model was used to evaluate the prognostic significance of adjuvant TACE. RESULTS: In the patients without any risk factors for the residual tumor, the 1-, 3-, 5-year survival rates were 93.48%, 75.85%, 62.39% in the control group and 97.39%, 70.37%, 50.85% in the adjuvant TACE group, respectively. There was no significant difference in the survival between two groups (P = 0.3956). However, in the patients with risk factors for residual tumor, postoperative adjuvant TACE significantly prolonged the patients' survival. There was a statistically significant difference in survival between two groups (P = 0.0216). The 1-, 3-, 5-year survival rates were 69.95%, 49.86%, 37.40% in the control group and 89.67%, 61.28%, 44.36% in the adjuvant TACE group, respectively. Cox proportional hazard model showed that tumor diameter and cirrhosis, but not the adjuvant TACE, were the significantly independent prognostic factors in the patients without risk factors for residual tumor. However, in the patients with risk factors for residual tumor adjuvant TACE, and also tumor diameter, AFP level, vascular invasion, were the significantly independent factors associated with the decreasing risk for patients' death from HCC. CONCLUSION: Postoperative adjuvant TACE can prolong the survival of patients with risk factors for residual tumor, but can not prolong the survival of patients without risk factors for residual tumor. PMID- 15334672 TI - Heparanase mRNA expression and point mutation in hepatocellular carcinoma. AB - AIM: To explore the expression of heparanase mRNA and point mutation in hepatocellular carcinoma (HCC). METHODS: Reverse transcription polymerase chain reaction was used to measure the expression of heparanase mRNA in the primary tumor tissues and surrounding liver tissues of 33 HCC patients. T-A cloning and sequencing were used to detect whether there was any mutation in the amplified PCR products. RESULTS: The expression of heparanase mRNA was positive in 16 primary tumor tissues of HCC, and the positive rate was 48.5%, which was significantly higher than that in the surrounding liver parenchyma (P<0.01). The positive rate for heparanase gene in high-tendency to metastatic recurrence group (71.4%, 10/14) was obviously higher than that in low-tendency to metastatic recurrence group (31.6%, 6/19) (P = 0.023). The positive rate for heparanase gene in patients with metastatic recurrence during postoperative follow-up (78.6%, 11/14) was also significantly higher than that in those without metastatic recurrence (21.4%, 3/14) (P = 0.003). Sequence analysis of the HPA PCR products was made in 7 patients, and 2-point mutations were found in 4 patients, one of which was sense mutation, neither base insertion nor deletion was detected. The mutation rate was 57.1% (4/7). CONCLUSION: The expression rate of heparanase mRNA increases in HCC, and HPA mRNA may be one of the reliable markers for the metastatic activity gained by the liver tumor cells and could be used clinically in predicting metastatic recurrence of HCC. Point mutation may be one of the causes for enhanced heparanase mRNA expression. PMID- 15334673 TI - Lymphatic metastasis and nm23H1 genetic instability in Chinese colon cancer patients. AB - AIM: To investigate the pathogenic mechanism of colon cancer at the molecular level and to elucidate the relationship between intercellular adhesion molecule-1 (ICAM-1) and nm23H(1) genes and Chinese patients with colon cancer. METHODS: DNA was extracted from paraffin-embedded materials. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze MSI and LOH. Expression of ICAM-1 was detected by Envision immuno-histochemistry. Experimental results were analyzed with Leica-Qwin computer imaging techniques and SPSS software of statistics. RESULTS: ICAM-1 expression of lymphatic endothelium was negative in normal colon and positive in colon cancer respectively. The number of lymphatics positive for ICAM-1 was gradually increased with degree of cancer invasion (P<0.01). In the group with metastasis of colon cancer, the number of lymphatics positive for ICAM-1 in lymph nodes was more than that in the group with no metastasis (P<0.01). The frequency of MSI, LOH and nm23H(1) protein was 26.67%, 20.00% and 53.33% in colon cancer, respectively. In TNM staging, MSI (43.75%) and nm23H(1) protein (81.25%) in stages I+II were detected more easily than the corresponding indexes (MSI: 7.14%, P<0.05 and nm23H(1): 21.43%, P<0.01) in stages III+IV. By comparison, the frequency of LOH (35.71%) in stages III+IV was more than that of LOH (6.25%, P<0.05) in stages I+II. LOH exhibited a rising trend along with the Duke's staging. nm23H(1) protein in the group of tubular adenocarcinoma (60.00%) was higher expressed than that in the group of mucoid adenocarcinoma (20.00%) (P<0.01), and exhibited a rising trend with the differentiation degrees of tubular adenocarcinoma. nm23H(1) protein in MSI positive group was higher expressed (75%) than that in MSI negative group (45.45%, P<0.05). CONCLUSION: The expression of ICAM-1 in lymphatic vessels is beneficial to the judgement of the invasion and metastasis ability of colon cancer and the anti-tumor immunity function, and shows an important clinical significance in predicting lymphatic metastasis of colon cancer. MSI and LOH may separately control the development of sporadic colon cancer with different pathways. LOH mostly arises in the late period of sporadic colon cancer and endows a high aggressive and poor prognostic phenotype. By compassion, MSI may be an early period molecule marker for sporadic colon cancer, enhanced expression of nm23H(1) protein can effectively inhibit colon cancer metastasis and improve prognosis of sporadic colon cancer patients. PMID- 15334674 TI - Cytochrome C oxidase III interacts with hepatitis B virus X protein in vivo by yeast two-hybrid system. AB - AIM: To screen and identify the proteins which interact with hepatitis B virus (HBV) X protein in hepatocytes by yeast two-hybrid system and to explore the effects of X protein in the development of hepatocellular carcinoma (HCC). METHODS: With HBV X gene amplified by polymerase chain reaction (PCR), HBV X bait plasmid, named pAS2-1-X, was constructed by yeast-two hybridization system3 and verified by auto-sequencing assay. pAS2-1-X was transformed into the yeast AH109, and X-BD fusion protein expressed in the yeast cells was detected by Western blotting. The yeast cells cotransformed with pAS2-1-X and normal human liver cDNA library were grown in selective SC/-trp-leu-his-ade medium. The second screen was performed with beta-gal activity detection, and false positive clones were eliminated by segregation analysis, true positive clones were amplified, sequenced and analyzed with bioinformatics. Mating experiment was performed to confirm the binding of putative proteins to X protein in the yeast cells. RESULTS: Bait plasmid pAS2-1-X was successfully constructed and pAS2-1-X correctly expressed BD-X fusion protein in yeast AH109. One hundred and three clones grew in the selective SC/-trp-leu-his-ade medium, and only one clone passed through beta-gal activity detection and segregation analysis. The inserted cDNA fragment showed high homology with Homo sapiens cytochrome C oxidase III (COXIII). Furthermore, mating experiment identified that the binding of COXIII to X protein was specific. CONCLUSION: COXIII protein is a novel protein that can interact with X protein in vivo by yeast two-hybrid system, and may contribute to the development of HCC through the interaction with X protein. PMID- 15334675 TI - Roles of Helicobacter pylori infection and cyclooxygenase-2 expression in gastric carcinogenesis. AB - AIM: Cyclooxygenase (COX)-2 is over expressed in gastrointestinal neoplasm. Helicobacter pylori (H pylori) infection is causally linked to gastric cancer. However, the expression of COX-2 in various stages of H pylori-associated gastric carcinogenesis pathway has not been elucidated. Therefore, the aim of this study was to clarify the role of H pylori induced COX-2 expression during carcinogenesis in the stomach. METHODS: Gastric biopsies from 138 subjects (30 cases of chronic superficial gastritis (CSG), 28 cases of gastric glandular atrophy (GA), 45 cases of gastric mucosal intestinal metaplasia (IM), 12 cases of moderate gastric epithelial dysplasia and 23 cases of gastric cancer) were enrolled. H pylori infection was assessed by a rapid urease test and histological examination (modified Giemsa staining). The expression of COX-1 and COX-2 in human gastric mucosa was detected by immunohistochemical staining. RESULTS: H pylori infection rate was 64.3% in GA and 69.5% in gastric cancer, which was significantly higher than that (36.7%) in CSG (P<0.05). The positive expression rates of COX-2 were 10.0%, 35.7%, 37.8%, 41.7% and 69.5% in CSG, GA, IM, dysplasia and gastric cancer, respectively. From CSG to GA, IM, dysplasia and finally to gastric cancer, expression of COX-2 showed an ascending tendency, whereas COX-1 expression did not change significantly in the gastric mucosa. The level of COX-2 expression in IM and dysplasia was significantly higher in H pylori-positive than in H pylori-negative subjects (P<0.01). CONCLUSION: COX-2 expression induced by H pylori infection is a relatively early event during carcinogenesis in the stomach. PMID- 15334676 TI - Lansoprazole ameliorates intestinal mucosal damage induced by ischemia reperfusion in rats. AB - AIM: To investigate the protective effect of lansoprazole on ischemia and reperfusion (I/R)-induced rat intestinal mucosal injury in vivo. METHODS: Intestinal damage was induced by clamping both the superior mesenteric artery and the celiac trunk for 30 min followed by reperfusion in male Sprague-Dawley rats. Lansoprazole was given to rats intraperitoneally 1 h before vascular clamping. RESULTS: Both the intraluminal hemoglobin and protein levels, as indices of mucosal damage, significantly increased in I/R-groups comparison with those of sham-operation groups. These increases in intraluminal hemoglobin and protein levels were significantly inhibited by the treatment with lansoprazole at a dose of 1 mg/kg. Small intestine exposed to I/R resulted in mucosal inflammation that was characterized by significant increases in thiobarbituric acid-reactive substances (TBARS), tissue-associated myeloperoxidase activity (MPO), and mucosal content of rat cytokine-induced neutrophil chemoattractant-1 (CINC-1). These increases in TBARS, MPO activities and CINC-1 content in the intestinal mucosa after I/R were all inhibited by pretreatment with lansoprazole at a dose of 1 mg/kg. Furthermore, the CINC-1 mRNA expression was increased during intestinal I/R, and this increase in mRNA expression was inhibited by treatment with lansoprazole. CONCLUSION: Lansoprazole inhibits lipid peroxidation and reduces development of intestinal mucosal inflammation induced by I/R in rats, suggesting that lansoprazole may have a therapeutic potential for I/R injury. PMID- 15334677 TI - Hepatic differentiation capability of rat bone marrow-derived mesenchymal stem cells and hematopoietic stem cells. AB - AIM: To investigate the different effects of mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) on hepatic differentiation. METHODS: MSCs from rat bone marrow were isolated and cultured by standard methods. HSCs from rat bone marrow were isolated and purified by magnetic activated cell sorting. Both cell subsets were induced. Morphology, RT-PCR and immunocytochemistry were used to identify the hepatic differentiation grade. RESULTS: MSCs exhibited round in shape after differentiation, instead of fibroblast-like morphology before differentiation. Albumin mRNA and protein were expressed positively in MSCs, without detection of alpha-fetoprotein (AFP). HSCs were polygonal in shape after differentiation. The expression of albumin signal decreased and AFP signal increased. The expression of CK18 was continuous in MSCs and HSCs both before and after induction. CONCLUSION: Both MSCs and HSCs have hepatic differentiation capabilities. However, their capabilities are not the same. MSCs can differentiate into mature hepatocyte-like cells, never expressing early hepatic specific genes, while Thy-1.1(+) cells are inclined to differentiate into hepatic stem cell-like cells, with an increasing AFP expression and a decreasing albumin signal. CK18 mRNA is positive in Thy-1.1(+) cells and MSCs, negative in Thy-1.1( ) cells. It seems that CK18 has some relationship with Thy-1.1 antigen, and CK18 may be a predictive marker of hepatic differentiation capability. PMID- 15334678 TI - Effects of Zuogui Wan on neurocyte apoptosis and down-regulation of TGF-beta1 expression in nuclei of arcuate hypothalamus of monosodium glutamate -liver regeneration rats. AB - AIM: To inquire into the effects and mechanism of Zuogui Wan (Pills for Kidney Yin) on neurocyte apoptosis in nuclei of arcuate hypothalamus (ARN) of monosodium glutamate (MSG)-liver regeneration rats, and the mechanism of liver regeneration by using optic microscope, electron microscope and in situ end labeling technology to adjust nerve-endocrine-immunity network. METHODS: Neurocyte apoptosis in ARN of the experiment rats was observed by using optic microscope, electron microscope and in situ end labeling technology. Expression of TGF-beta1 in ARN was observed by using immunohistochemistry method. RESULTS: The expression of TGF-beta1 in rats of model group was increased with the increase of ARN neurocyte apoptosis index (AI) (t = 8.3097, 12.9884, P<0.01). As compared with the rats of model group, the expression of TGF-beta1 in rats of Zuogui Wan treatment group was decreased with the significant decrease of ARN neurocyte apoptosis (t = 4.5624, 11.1420, P<0.01). CONCLUSION: Brain neurocyte calcium ion overexertion and TGF-beta1 protein participate in the adjustment and control of ARN neurocyte apoptosis in MSG-liver regeneration-rats. Zuogui Wan can prevent ARN neurocyte apoptosis of MSG-liver regeneration in rats by down-regulating the expression of TGF-beta1, and influence liver regeneration through adjusting nerve endocrine-immune network. PMID- 15334679 TI - Autocrine expression of hepatocyte growth factor and its cytoprotective effect on hepatocyte poisoning. AB - AIM: To construct pEGFP-hepatocyte growth factor (HGF) expression vector, the to detect its expression in transfected human hepatocytes, and to investigate the influence of autocrine HGF expression on the proliferative potential and cytoprotective effects in human hepatocytes. METHODS: Human HGF cDNA was ligated to the pEGFP vector. Recombinant plasmid was transfected into human hepatocyte line QZG with liposome. Expression of HGF protein was observed by fluorescence microscopy and immunohistochemistry. Hepatic cells were collected 24, 48, and 72 h after transfection to detect the number of ((3)H)-TdR uptake in DNA. DNA synthesis was observed by using PCNA stain immunohistochemistry. Acute liver cell damage was induced by carbon tetrachloride. Cytoprotective effect was observed by examining the survival rate of hepatocytes and leakage of intracellular alanine transaminase (ALT) and potassium ions. RESULTS: HGF identification of pEGFP-HGF by enzyme digestion showed that HGF fragment was cloned into BamH I and Sal I sites of pEGFP-N3. Expression of GFP in transfected hepatocytes was observed with fluorescence microscopy. The ((3)H)-TdR uptake became 7 times as many as in the control group 96 h after transfection. After HGF transfection, the survival rate of hepatocytes poisoned by CCl(4) significantly increased (83% vs 61%, P<0.05), and the leakage of intracellular alanine transaminase and potassium ions decreased (586 nkat/L vs 1 089 nkat/L, P<0.01; and 5.59 mmol/L vs 6.02 mmol/L, P<0.01 respectively). Culture of transfected hepatic cells promoted the proliferation of other non-transfected cells. CONCLUSION: Transfected HGF is expressed in hepatic cells and has the activity of promoting cell division and protecting hepatic cells against poisoning. PMID- 15334680 TI - Effects of herbal compound 861 on human hepatic stellate cell proliferation and activation. AB - AIM: To investigate the effects of herbal compound 861 (Cpd 861) on cell proliferation in human hepatic stellate cells (LX-2) and human hepatocellular liver carcinoma cells (HepG2), and expression of alpha-smooth muscle actin (alpha SMA) in LX-2 cells. METHODS: LX-2 and HepG2 cells were incubated with various concentrations of Cpd 861 (0.1-0.003 mg/mL) for 1, 2, 3, 5 and 7 d. Cell proliferation was analyzed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Effects of Cpd861 on the expression of alpha-SMA mRNA in LX-2 cells were measured by real time quantitative PCR method using SYBR Green I technology. RESULTS: Cpd 861, at 0.1 mg/mL, significantly inhibited LX-2 cell proliferation (15% decrease relative to control, P<0.05) after 3 d of incubation. The inhibitory effects seemed to increase with the treatment time (25% decrease after 5 d of incubation and 35% decrease after 7 d of incubation, P<0.01). However, Cpd 861 did not affect HepG2 cell proliferation at the same concentration used for LX-2 cells. The expression levels of alpha-SMA mRNA decreased significantly when LX-2 cells were exposed to Cpd 861 for 48 h (59% decrease relative to control, P<0.05) or 72 h (60% decrease relative to control, P<0.01). CONCLUSION: Cpd 861 can significantly inhibit LX-2 cell proliferation in a dose-dependent manner, and reduce the expression levels of alpha-SMA mRNA in LX-2 cells. Since hepatic cell proliferation and high level of alpha-SMA are associated with liver fibrosis, the results suggest that Cpd 861 may be useful in the treatment of this disease. PMID- 15334681 TI - Tegaserod inhibits noxious rectal distention induced responses and limbic system c-Fos expression in rats with visceral hypersensitivity. AB - AIM: To examine the effects of tegaserod, a serotonin (5-HT) 4 receptor partial agonist, on abdominal withdrawal reflex (AWR) to rectal distention (RD) and c-Fos expression in limbic system. METHODS: Neonatal Sprague-Dawley rats randomly received colonic irritation by acetic acid from postnatal day 8 to d 21 as a visceral hypersensitive model (group H) or by intrarectal saline as a control group (group C). When they became adults, rectal distention (RD) was performed by a balloon (6F; Fogarty arterial embolectomy catheter; length, 20 mm; diameter, 2 mm) which was rapidly inflated with increasing volumes of saline (0.4, 0.8 and 1.2 mL) for 20 s at five-minute intervals. Five subgroups of group H (H-saline, H vehicle, H-Teg0.1, H-Teg0.3 and H-Teg1.0) were injected randomly with saline, vehicle (1-methyl-2-thpyrrolidone) or tegaserod at doses of 0.1, 0.3 and 1.0 mg/kg ip, respectively. Two subgroups of group C (C-Saline and C-Teg1.0) were injected with saline or tegaserod (1.0 mg/kg) ip. RD was performed 10 min after injection, AWR was recorded and c-Fos expression in limbic system was analyzed quantitatively by immunohistochemistry. RESULTS: Compared to saline, tegaserod significantly inhibited AWR in group H (0.4 mL: from 2.0 to 0.5; 0.8 mL: from 3.5 to 1.5; 1.2 mL: from 4.0 to 3.0, P<0.01), but had no significant effect on group C. Tegaserod dose-dependently attenuated the number of c-Fos positive neurons in limbic structures, anterior cingulate cortex (ACC) showed the greatest attenuation. In group H, tegaserod (1.0 mg/kg) resulted in a significant overall decrease to 57% of H-saline (283+/-41 vs 162+/-16, P<0.01), in ACC to 42% of H saline (72+/-10 vs 31+/-8, P<0.01). In group C, tegaserod (1.0 mg/kg) resulted in an overall decrease to 77% of C-saline (214+/-13 vs 164+/-22, P<0.01), in ACC to 65% of C-saline (48+/-8 vs 31+/-7, P<0.01). CONCLUSION: Tegaserod inhibits the response to rectal distention in rats with visceral hypersensitivity and dose dependently attenuates c-Fos expression in limbic system, especially in anterior cingulate cortex. PMID- 15334682 TI - Inhibitory effects of berberine on ion channels of rat hepatocytes. AB - AIM: To examine the effects of berberine, an isoquinoline alkaloid with a long history used as a tonic remedy for liver and heart, on ion channels of isolated rat hepatocytes. METHODS: Tight-seal whole-cell patch-clamp techniques were performed to investigate the effects of berberine on the delayed outward potassium currents (I(K)), inward rectifier potassium currents (I(K1)) and Ca(2+) release-activated Ca(2+) currents (I(CRAC)) in enzymatically isolated rat hepatocytes. RESULTS: Berberine 1-300 micromol/L reduced I(K) in a concentration dependent manner with EC(50) of 38.86+/-5.37 micromol/L and n(H) of 0.82+/-0.05 (n = 8). When the bath solution was changed to tetraethylammonium (TEA) 8 mmol/L, I(K) was inhibited. Berberine 30 micromol/L reduced I(K) at all examined membrane potentials, especially at potentials positive to +60 mV (n = 8, P<0.05 or P<0.01 vs control). Berberine had mild inhibitory effects on I(K1) in rat hepatocytes. Berberine 1-300 micromol/L also inhibited I(CRAC) in a concentration-dependent fashion. The fitting parameters were EC(50) = 47.20+/-10.86 micromol/L, n(H) = 0.71+/-0.09 (n = 8). The peak value of I(CRAC) in the I-V relationship was decreased by berberine 30 micromol/L at potential negative to -80 mV (n = 8, P<0.05 vs control). But the reverse potential of I(CRAC) occurred at voltage 0 mV in all cells. CONCLUSION: Berberine has inhibitory effects on potassium and calcium currents in isolated rat hepatocytes, which may be involved in hepatoprotection. PMID- 15334683 TI - Surgical management of esophageal strictures after caustic burns: a 30 years of experience. AB - AIM: To analyze a 30-year historical series of patients treated in our hospital, who ingested corrosive substances, and to assess the effectiveness of surgical therapy administered in patients with strictures after caustic injury in esophagus during this period. METHODS: A total of 79 cases of caustic burns in esophagus were treated in Tangdu Hospital from 1971 to 2001. Their clinical and pathological data were reviewed, and collected from the medical records of patients and interviews with them. RESULTS: More men (n = 61) than women (n = 18) ingested caustic substances with a sex ratio of 3.4:1 during the 30-year period. The caustic materials were liquid lye and acids (54 cases and 25 cases, respectively). Sixty-eight patients were given esophageal replacement in more than three months after caustic injury with no postoperative death, of which 17 cases developed postoperative complications making a complication rate of 25%. The most common one was cervical anastomotic leakage. All patients had improvement in swallowing afterwards. CONCLUSION: The presence and severity of injuries are correlated with the amount of caustic substances ingested. Surgical treatment is a good option in patients with severe strictures, and colonic interposition might be the best surgical process. The most important factors to guarantee a successful outcome for surgery are good vascular supply and absence of tension in the anastomosis. PMID- 15334684 TI - Laparoscopic resection of submucosal tumor on posterior wall of gastric fundus. AB - AIM: Laparoscopic resection of tumors on the posterior wall of gastric fundus, especially when they are next to the esophagocardiac junction (ECJ), is both difficult and time-consuming. Furthermore, it can lead to inadvertent esophagus stenosis and injury to the spleen. In order to overcome these difficulties, laparoscopically extraluminal resection of gastric fundus was designed to manage submucosal tumors located on the posterior wall of gastric fundus and next to ECJ. METHODS: From January 2001 to September 2003, laparoscopically extraluminal resection of gastric fundus was successfully carried out on 15 patients. There were 11 males and 4 females with an average age of 58 years (range, 38 to 78 years). The mean diameter of the tumors was 4.8 cm. The distance of the tumor border from ECJ was about 1.5-2.5 cm. The four-portal operation procedures were as follows: localization of the tumor, dissection of the omentum, mobilization of the gastric fundus and the upper polar of spleen, exposure of ECJ, and resection of the gastric fundus with Endo GIA. RESULTS: The laparoscopic operation time averaged (66.2+/-10.4) min, the average amount of bleeding was (89.4+/-21.7) mL. The mean post-operative hospital stay was (5.3+/-1.1) d. Within 36 h post operation, 73.3% of all the patients recovered their gastrointestinal function and began to eat something and to walk. In all the operations, no apparent tumor focus was left and no complication or conversion to open surgery occurred. CONCLUSION: Our newly designed procedure, laparoscopically extraluminal resection of the gastric fundus, can avoid contamination of the abdominal cavity, injury to the spleen and esophageal stenosis. The procedure seems to be both safe and effective. PMID- 15334685 TI - First two cases of living related liver transplantation with complicated anatomy of blood vessels in Beijing. AB - AIM: Living related liver transplantation (LRLT) has been developed in response to the paediatric organ donor shortage. Though it has been succeeded in many centers worldwide, the safety of the donor is still a major concern, especially in donors with anatomy variation. We succeeded in performing the first two cases of living related liver transplantation with complicated anatomy of blood vessels as a way to overcome cadaveric organ shortage in Beijing. METHODS: Two patients, with congenital liver fibrosis and congenital biliary atresia were performed with living donor liver transplantation in our hospital and then followed up from November 12 to December 13, 2001. The two living donors, mother and father, were healthy aged 34 and 35 years. One right lobe (segment V, VI, VII, VIII) and one left lateral lobe (segment II and III) were used. The grafts weighed 394 g and 300 g. The ratio of graft weight to the standard liver volume (SLV) of donors was 68% and 27%. The graft weight to recipient body weight ratio was 3.2% and 4.4%. The graft weight to recipient estimated standard liver mass (ESLM) ratio was 63% and 85%. The two donors had complicated blood vessel variation. RESULTS: Two patients undergone living donor liver transplantation had good results. Abnormal liver function with high bilirubin level appeared in a few days after operation, but liver function returned to normal one month after operation with bilirubin level almost decreased to near normal. No bleeding, thrombosis, infection and bile leakage occurred. One had an acute rejection and recovered. The two donors recovered in two weeks. One had slight fever because of a little collection in abdomen and recovered after paracentesis and drainage. CONCLUSION: Living donor liver transplantation has been proved to be a good way that offers a unique opportunity of getting a timely liver graft as a response to shortage of pediatric donors, though it could be a technically difficult operation if there is anatomical variation. PMID- 15334686 TI - Transcutaneous perianal sonography: a sensitive method for the detection of perianal inflammatory lesions in Crohn's disease. AB - AIM: Pelvic magnetic resonance imaging (MRI) and endoanal ultrasound which are established imaging methods for perianal inflammatory lesions in patients with Crohn's disease require expensive specialized equipments and expertise. We investigated the feasibility and sensitivity of transcutaneous perianal ultrasound (PAUS) using regular ultrasound probes in the imaging of perianal inflammatory lesions. The sonographic findings were correlated to pelvic MRI scans. METHODS: We performed PAUS in 25 patients with Crohn's disease and clinical signs of perianal inflammatory disease. Within a median of 10 d (range 0 75) these patients underwent MRI of the pelvis. Regular convex and linear high resolution probes were used for PAUS. The sonographic findings were correlated to the MRI findings by blinded investigators. RESULTS: The sonographic investigations were well tolerated by all patients. Fistulae typically presented as hypoechoic tracks. Twenty-nine fistulae were detected in 22 patients. Abscesses were detected in 7 patients and presented as hypo- or anechoic formations. Twenty-six of 29 fistulae and 6 of 7 abscesses could be confirmed by MRI. Kappa statistics showed an excellent agreement (kappa>0.83) between the two imaging methods. CONCLUSION: PAUS is a simple, painless, feasible, real-time method that can be performed without specific patient preparation which is comparable in its sensitivity to pelvic MRI in the detection of perianal fistulae and/or abscesses. PAUS can especially be recommended as a screening tool in acute perianal disorders such as perianal abscess and for follow-up studies of perianal inflammatory disease. PMID- 15334687 TI - Low plasma apolipoprotein A-I level is not a reliable marker of fibrosis in children with chronic hepatitis B. AB - AIM: To evaluate the clinical value of plasma apolipoprotein A-I (Apo A-I) as a marker of fibrosis in children with chronic hepatitis B (CHB). METHODS: Liver biopsy specimens from 49 children with CHB were evaluated by using Knodell index. Plasma Apo A-I level was measured after 12-h fasting. Student's t test, Spearman's correlation test and receptor-operating characteristic (ROC) curve were used for statistical evaluation. RESULTS: Mean Apo A-I level of the patients was not different from that of controls (P>0.05). Six (8.7%) children had fibrosis score of more than 2 (severe fibrosis). No difference in the level of mean plasma Apo A-I was found among children with and without severe fibrosis (P>0.05). No correlation between Apo A-I level and fibrosis scores was found (P>0.05). The area under the ROC curve was 0.407+/-0.146 (P>0.05). CONCLUSION: Severe fibrosis is not common in children with CHB and plasma Apo A-I level is not a reliable indicator of fibrosis. PMID- 15334688 TI - Prospective study in 142 cases of hepatitis C virus infection. AB - AIM: There is limited information on the natural history of HCV infection in China. We investigated the outcome of HCV infection after nine-year follow-up and the risk factors in blood donors in China in order to provide the foundation for prevention and therapy. METHODS: A total of 172 cases of HCV infection with anti HCV positive and ALT abnormality were enrolled in the archives when was screened blood in Hebei Province in 1993. In them 142 blood donors were followed up till July 2002. No antiviral treatment was applied to them during the period of infection. In the present study, anti-HCV, HCV-RNA and aminotransferase were detected and genotyping was conducted by the method of restriction fragment length polymorphism(RFLP). B-type ultrasound detection was performed in all the patients. Age, sex, alcohol consumption and clinical symptoms were questioned. RESULTS: After nine years' follow-up, 10.56% (15/142) of the cases were negative for anti-HCV and 16.42% (12/134) of them were negative for HCV-RNA. The genotypes 1b, 2a and 1b/2a were 91.07%, 6.25% and 2.68% respectively. Twelve cases (8.45%) were negative for both HCV RNA and anti-HCV. The rate of chronicity in this group was 83.58% (112/134), and the rate of viral spontaneous resolution was 16.42% (22/134). The mean level of ALT, AST, gamma-GT in HCV RNA positive cases was significantly higher than that in HCV RNA negative cases (P<0.001). The abnormal rate of ALT and/or AST in male donors was significantly higher than that in female donors (P = 0.005). The rate of progression to liver cirrhosis from chronic hepatitis C was significantly higher in the cases of super-infection with HBV than that in the cases of single HCV infection. Overdose alcohol consumption promoted the progression to chronicity. CONCLUSION: This area (Hebei Province) has a higher rate of chronicity in HCV infection, and measures should be taken to prevent its progression to serious liver diseases, especially for patients super infected with HCV and HBV. PMID- 15334689 TI - Ability of luteinizing hormone releasing hormone-Pseudomonas aeruginosa exotoxin 40 binding to LHRH receptor on human liver cancer cells. AB - AIM: To explore the ability of recombinant toxin luteinizing hormone releasing hormone-Pseudomonas aeruginosa exotoxin 40 (LHRH-PE40) and LHRH binding to LHRH receptor (LHRHR) on the membrane surface of human liver cancer HEPG cells. METHODS: LHRH was labeled by using (125)I with enzymatic reaction. The affinity and receptor volume of LHRH-PE40 and LHRH binding to LHRHR on the membrane surface of human liver cancer cells were measured with radioligand receptor assay. RESULTS: The specific activity of LHRH labeled with (125)I was 2.7 x 10(4) kBq/microL, and its radiochemical purity reached to 99.2-99.7%. The binding of (125)I to LHRH was maximal for 240 min in the warm cultivation, and this binding was stabilized. The inhibiting rates of (125)I-LHRH and LHRH on the proliferation of human liver cancer HEPG cells were not significantly different. On the basis of the saturation curve of (125)I-LHRH binding to the membrane LHRHR of HEPG cells, (125)I-LHRH of 1 x 10(5) cpm was selected for radioligand receptor assay. The affinity constants (Kd) of LHRH-PE40 and LHRH binding to the membrane LHRHR of HEPG cells were 0.43+/-0.12 nmol/L and 4.86+/-1.47 nmol/L, respectively, and their receptor volumes were 0.37+/-0.15 micromol/g and 0.42+/-0.13 micromol/g, respectively. The binding of LHRH-PE40 to the membrane protein of normal liver cells was not observed. CONCLUSION: The recombinant toxin LHRH-PE40 binding to the membrane surface of LHRHR of human liver cancer HEPG cells was very strong, while the specific binding of it to normal liver cells was not observed. The results provide an important experimental basis for the clinical application of LHRH-PE. PMID- 15334690 TI - Human endostatin gene transfer, either naked or with liposome, has the same inhibitory effect on growth of mouse liver tumor cells in vivo. AB - AIM: To explore a safe and efficient strategy of tumor therapy using anti angiogenetic agents. METHODS: Endostatin gene with a signal sequence of human IgGgamma chain was amplified by PCR and cloned into pVAX1 plasmid which was the only vector authorized by FDA in clinical trial to construct a recombinant plasmid named as pVAX-sEN. The recombinant plasmid was detected with EcoRI/KpnI and DNA sequencing. BALB/c mice bearing hepatocarcinoma cell line H22 were treated with naked pVAX-sEN or liposome-DNA complex in which the dose of DNA and the ratio of DNA and liposome were different from each other. To compare the efficiency of gene transfection, expression of endostatin at the treated tumor site was assayed with ELISA. To investigate the effect of pVAX1-sEN on hepatocellular carcinoma, pVAX-sEN either naked or in liposome-DNA complex was injected into BALB/c mice bearing H22, then the diameter of tumors was measured, microvessel density was detected by immunohistochemistry, endostatin expression in vivo was assayed at different time points. RESULTS: DNA sequencing showed the endostatin gene with the signal peptide was correctly cloned. In situ gene expression assay indicated that both the ratio of DNA and liposome and the dose of DNA could affect the gene transfection efficiency. Interestingly, naked pVAX sEN had a similar in situ endostatin expression to pVAX-sEN with liposome. Animal experiments showed that pVAX-sEN together with pVAX-sEN-liposome complex could efficiently suppress the growth of mouse hepatoma cells. CONCLUSION: Naked endostatin plasmid intratumoral injection can get a similar gene transfection efficiency to liposome-DNA complex when used in situ. PMID- 15334691 TI - Expression of plasma vascular endothelial growth factor in patients with hepatocellular carcinoma and effect of transcatheter arterial chemoembolization therapy on plasma vascular endothelial growth factor level. AB - AIM: To investigate the expression level of plasma vascular endothelial growth factor (P-VEGF) in patients with hepatocellular carcinoma (HCC) and its relationship with the clinicopathologic characteristics, and to examine the changes of P-VEGF in the course of transcatheter arterial chemoembolization (TACE). METHODS: Peripheral blood samples were taken from 45 HCC patients before and 1, 3, 7 d, and 1 mo after TACE. Plasma VEGF level was measured with the quantitative sandwich enzyme-linked immunosorbent assay (ELISA). Twenty patients with benign liver lesions and 17 healthy control subjects were also included in this study. RESULTS: Plasma VEGF levels in HCC patients were significantly elevated as compared to those in patients with benign liver lesions (P = 0.006) and in the normal controls (P = 0.003). Significant differences were observed when P-VEGF was categorized by tumor size (P = 0.006), portal vein thrombosis (P = 0.011), distant metastasis (P = 0.017), arterial-portal vein shunting (P = 0.026), and International Union Against Cancer (UICC) TNM stage (P = 0.044). There was no correlation between plasma level of VEGF and the level of alpha fetoprotein (alpha-FP) (r = 0.068, P = 0.658) and weakly correlated with the number of platelets (r = 0.312, P = 0.038). P-VEGF levels increased significantly and reached the peak value on the first day after TACE, and then decreased gradually. The change rate of P-VEGF concentration (one month post-TACE/pre TACEX100%) was correlated with the retention rate of lipiodol oil (r s = 0.494, P = 0.001) and the tumor volume change (r s = 0.340, P = 0.034). The patients who achieved a partial or complete response to TACE therapy showed significantly less pre-treatment P-VEGF than those nonresponders (P = 0.025). A high pre-therapeutic P-VEGF level was associated with poor response to treatment (P = 0.018). CONCLUSION: A high pre-treatment P-VEGF level is a useful marker for tumor progression, especially for vascular invasion. TACE increases the level of P-VEGF only temporarily which may be associated with tumor ischemia. P-VEGF may be useful in predicting treatment response, monitoring disease course after TACE and judging the effect of different TACE regimens. PMID- 15334692 TI - Quantifying anti-HBV effect of targeted ribonuclease by real-time fluorescent PCR. AB - AIM: To quantify the inhibition of HBV replication by targeted ribonuclease by using real-time fluorescent PCR. METHODS: Targeted ribonuclease was introduced into 2.2.15 cells by liposome-mediated transfection or HIV-TAT mediated protein transduction. Forty-eight hours after the transfection and 24 h after the transduction, supernatants of 2.2.15 cells were collected and HBV DNA in the supernatants was quantified by real-time fluorescent PCR with a commercial kit. RESULTS: HBV DNA concentrations in the supernatants of 2.2.15 cells transfected or transducted with targeted ribonuclease were 4.9+/-2.4 x 10(8) copies/L and 8.3+/-4.0 x 10(8) copies/L, respectively. Compared with controls, transfection or transduction of targeted ribonuclease reduced HBV DNA concentration in the supernatants of 2.2.15 cells by 90.4% and 90.1%, respectively (P<0.05). CONCLUSION: Targeted ribonuclease can inhibit HBV replication in 2.2.15 cells. PMID- 15334693 TI - Expression of survivin and its significance in colorectal cancer. AB - AIM: To study the expression of survivin, a novel member of inhibitors of apoptosis protein (IAP) and its significance in colorectal carcinoma. METHODS: Survivin mRNA expression was evaluated by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) in 52 colorectal carcinoma samples and 48 adjacent normal colorectal tissue samples. PCR product was sequenced to verify the desired result. Expressions of survivin protein, proliferating cell nuclear antigen labelling index (PI) and apoptotic index (AI) were detected immunohistochemically in 52 human colorectal carcinomas. RESULTS: The expression of survivin mRNA was detected in a significantly greater proportion of colorectal carcinoma samples than in adjacent normal colorectal tissues (67.3% vs 25%; P<0.01). There was no relationship between survivin mRNA expression in colorectal carcinomas and sex, tumor size, histological types, lymphnode metastasis, distant metastasis and Dukes' stage. The PCR product shared 99% of homology with human counterparts. Survivin expression was observed immunohistochemically in 27 of 52 cases of colorectal carcinoma (51.9%). The AI was significantly lower in survivin positive group than in survivin negative group (0.67+/-0.18% vs 1.14+/-0.42%; P<0.001), while the PI was greater in survivin positive group than in survivin negative group (51+/-22% vs 27+/-18%, P<0.001). CONCLUSION: Survivin is a special tumor marker independent of histopathological characteristics. It may play an important role during human colorectal tumorigenesis by inhibiting apoptosis and accelerating proliferative activity of colorectal tumor cells. PMID- 15334694 TI - Expression of toll-like receptor 4 and MD-2 gene and protein in Kupffer cells after ischemia-reperfusion in rat liver graft. AB - AIM: To investigate the expression of toll-like receptor 4 (TLR4) and MD-2 gene and protein in Kupffer cells (KCs) and their role in ischemia-reperfusion (IR) injury of rat liver graft. METHODS: KCs were isolated at 0 (control group), 2, 12, 24 h (IR group) following IR in rat liver graft. mRNA expression of TLR4 and MD-2 was detected by RT-PCR analysis, protein expression of TLR4/MD-2 was detected by flow cytometric (FCM) analysis, and tumor necrosis factor-alpha (TNF alpha) level in supernatant was measured by ELISA. Then isolated KCs were incubated with anti-TLR4 polyclonal antibody (anti-TLR4 group), and TNF-alpha level was measured again. RESULTS: The mRNA and protein expression of TLR4/MD-2 and the level of TNF-alpha in IR group increased significantly at 2 h following IR, and reached the maximum at 12 h, and slightly decreased at 24 h, but were still significantly higher than those in the control group (P<0.01). The expression of these factors was markedly decreased after anti-TLR4 antibody treatment as compared with the IR group (P<0.01). CONCLUSION: Lipopolysaccharide (LPS) following IR can up-regulate TLR4/MD-2 gene and protein expression in KCs, and synthesize cytokine TNF-alpha. Anti TLR4 antibody can inhibit the production of TNF-alpha induced by LPS. TLR4 and its partner molecule MD-2 may play an important role in Kupffer cell activation and IR injury. PMID- 15334695 TI - Protective effects of recombinant human growth hormone on cirrhotic rats. AB - AIM: To investigate the effects and molecular mechanisms of recombinant human growth hormone (rhGH) on protecting liver function and alleviating portal hypertension of liver cirrhotic rats. METHODS: Liver cirrhosis of male Sprague Dawley rats was induced by administration of thioacetamide. The rats with or without liver cirrhosis were randomly divided into four groups. Group A consisted of the normal rats was treated with normal saline (NS), group B consisted of the normal rats was treated with rhGH, group C consisted of cirrhotic rats was treated with NS, and group D consisted of cirrhotic rats was treated with rhGH. The rats of different groups were subcutaneously injected with 0.5 mL of NS or 333 ng/kg of rhGH daily for 7 d. After treatments, the following parameters were examined, including GH-binding capacity (R(T)) by (125)I-hGH binding, growth hormone receptor mRNA(GHR mRNA) expression by RT-PCR, relative content of collagen (RCC) by histomorphomertry, and level of malon-dialdehyde (MDA) and superoxide dismutase (SOD) in liver tissue by thiobarbituric acid reaction and pyrogallic acid self-oxidation, respectively. Serum albumin (ALB), alanine transaminase (ALT) and portal vein pressure (PVP) were also examined. RESULTS: rhGH up-regulated both the GH-binding capacity (R(T)) and the expression of GHR mRNA in vivo. R(T) in group A (72+/-12 fmol/mg protein) was significantly higher than that in group C (31+/-4 fmol/mg protein) (P<0.05). R(T) in group B (80+/-9 fmol/mg protein) increased markedly compared to group A (P<0.05). R(T) in group D (40+/-7 fmol/mg protein) raised remarkably compared with group C (P<0.05), but less than that in group A, and there was no significant GH binding affinity contrast (Kd) change. The GHR mRNA level (iOD, pixel) in group A (29+/-3) was significantly higher than that in group C (23+/-3) (P<0.05). GHR mRNA levels were significantly raised in group B (56+/-4) and group D (42+/-8) compared with groups A and C (29+/-3 and 23+/-3, respectively) (P<0.05). Compared with the normal liver, MDA level was higher and SOD level was lower in cirrhotic livers. After rhGH treatment, MDA level was significantly declined to 12.0+/-2.2 nmol/mg protein and SOD was raised to 1 029+/-76 U/mg protein in group D (P<0.05). ALB levels in groups B and D (42+/-7 g/L and 37+/-7 g/L, respectively) were significantly raised compared with those in groups A and C (35+/-5 g/L and 29+/-4 g/L, respectively) (P<0.05). ALT level was markedly lower in group D (69+/-7 U/L) compared to group C (89+/-15 U/L) (P<0.05), and close to group A (61+/-10 U/L). RCC in group C (22.30+/-3.86%) was significantly higher than that in group A (1.14+/-0.21%) and group D (14.70+/-2.07%) (P<0.05). In addition, rhGH markedly alleviated portal hypertension in liver cirrhotic rats (group D vs C, 9.3+/-1.5 cmH(2)O vs 14.4+/-2.0 cmH(2)O) (P<0.05). CONCLUSION: Pharmacological doses of rhGH can increase R(T) and GHR mRNA expression, ameliorate liver functions, repress fibrosis and decline portal hypertension, suggesting it has potentially clinical usage as a hepatotropic factor. PMID- 15334696 TI - Tumor necrosis factor alpha antibody prevents brain damage of rats with acute necrotizing pancreatitis. AB - AIM: To study the protective effects of tumor necrosis factor alpha (TNFalpha) antibody on pancreatic encephalopathy in rats. METHODS: One hundred and twenty SD rats were randomly divided into normal control group, acute necrotizing pancreatitis group and TNFalpha antibody treated group. Acute hemorrhage necrotizing pancreatitis model in rats was induced by retrograde injection of 50 g/L sodium taurocholate into the pancreatobiliary duct. Serum TNFalpha was detected and animals were killed 12 h after drug administration. Changes in content of brain water, MDA and SOD as well as leucocyte adhesion of brain microvessels were measured. RESULTS: In TNFalpha antibody treated group, serum TNFalpha level was decreased. Content of brain water, MDA and SOD as well as leucocyte adhesion were decreased significantly in comparison with those of acute necrotizing pancreatitis group (P<0.05). CONCLUSION: TNFalpha antibody can alleviate the brain damage of rats with acute hemorrhage necrotizing pancreatitis. PMID- 15334697 TI - Effects of narcotic analgesic drugs on human Oddi's sphincter motility. AB - AIM: To assess the effects of intramuscular analgesics (morphine, Ap-237, pethidine and tramadol) on human Oddi's sphincter motility with choledochoscope manometry. METHODS: A total of 70 patients having T tubes after cholecystectomy and choledochotomy were assessed by choledochoscope manometry. They were randomly divided into morphine group, Ap-237 group, pethidine group and tramadol group. Basal pressure of Oddi's sphincter (BPOS), amplitude of phasic contractions (SOCA), frequency of phasic contractions (SOF), duration of phasic contractions(SOD), duodenal pressure (DP) and common bile duct pressure (CBDP) were scored and analyzed. All narcotic analgesic drugs were administered intramuscularly. RESULTS: Levels of BPOS, SOCA and SOF were increased after injection of morphine and Ap-237 (P<0.05), level of CBDP was increased from 4.97+/-3.87 mmHg to 8.62+/-7.43 mmHg (10 min later) and 7.32+/-5.95 mmHg (20 min later) after injection of morphine (P<0.01). No apparent change occurred after intramuscular injection of pethidine. Level of BPOS was increased from 7.01+/ 5.50 mmHg to 2.87+/-2.78 mmHg 10 min after injection of tramadol and SOCA was decreased from 63.34+/-35.29 mmHg to 45.90+/-27.86 mmHg (10 min later,P<0.05) and 35.97+/-24.30 (20 min later,P<0.01) after administration of tramadol. CONCLUSION: All these findings indicate that Oddi's sphincter manometry via choledochoscope is a practical and new way to study the dynamics of Oddi' s sphincter. The regular dose of morphine and Ap-237 could increase BPOS, SOF and SOCA. Morphine could increase the level of CBDP, demonstrating an excitatory effect on the sphincter of Oddi. Pethidine had no effect on Oddi's sphincter motility. Tramadol shows an inhibitory effect on the motility of the sphincter of Oddi and decreases levels of BPOS and SOCA. PMID- 15334698 TI - Angiographic findings of gastrointestinal stromal tumor. AB - AIM: To discuss the angiographic features of gastrointestinal stromal tumor (GIST) and to evaluate their diagnostic role. METHODS: Twelve patients with pathologically proved GIST underwent angiography (DSA)1 wk before operation, using Puck and digital subtraction DSA. The origin, size, morphology and angiographic appearance of the lesions were reviewed. RESULTS: Two tumors arose from stomach, 8 from jejunum, and 2 from ileum. Seven cases were benign and 5 were malignant. Obviously thickened supplying arteries were detected in 8 tumors, and early-developed veins were found in 3. Two types of angiographic changes of GIST were observed. Four cases had twisted irregular neoplastic vessels with partially coarse and indistinct margins, which were all malignant. Eight cases had ball-like neoplastic vessels with uniform tumor staining, of which 7 were benign and 1 was malignant. CONCLUSION: Angiography facilitates localization and diagnosis of GIST, helps define their size, range and location, and is especially valuable to patients suffering from melena with unknown reasons. PMID- 15334699 TI - Expression of growth hormone and its receptor in chronic atrophic gastritis and its clinical significance. AB - AIM: To investigate the growth hormone (GH) and growth hormone receptor (GHR) expression of and its clinical significance in patients with chronic atrophic gastritis (CAG). METHODS: A total of 90 cases were enrolled in the study. Thirty were healthy controls, the other 60 patients were divided into two groups according to the endoscopical and histological diagnosis. Blood samples were drawn in the morning (menarche did not occur during the blood extraction in female patients), gastric mucosa was obtained by endoscopy. Serum GH and gastrice mucosal GHR levels were measured using radioimmunoassay (RIA) and En Vinsion technique. RESULTS: The average GH level was 1.021+/-0.132 microg/L in CAG patients, in controls it was 2.869+/-0.512 microg/L. There was a significant difference between these two groups (P<0.01). The positive rate of GHR in CAG patients was 10%, in controls the rate was 100%. There was a significant difference (P<0.01). There was no significant change of GH level (3.176+/-0.421 microg/L) in patients with gastric carcinoma compared with controls (P>0.05). CONCLUSION: The study shows that levels of GH and GHR expression are low in CAG patients. CAG pathogenesis has a correlation with mucosal nutrient deficiency, decreased levels of GH and GHR have an adverse effect on the repair and regeneration of CAG. There is no significant change of GH in gastric carcinorma patients, GH dose not play a role in the pathogenesis of gastric cancer. PMID- 15334700 TI - Clinical and experimental study of oxaliplatin in treating human gastric carcinoma. AB - AIM: To evaluate the therapeutic effectiveness of oxaliplatin on human gastric carcinoma and to explore its mechanisms. METHODS: Twenty-two cases of stage IV gastric carcinoma received 4-6 (mean 4.6) cycles of first line combined chemotherapy with oxaliplatin (oxaliplatin 85 mg/m(2), iv, gtt, 1 h, d 1; leukovorin 200 mg/m(2), iv, gtt, 1 h, d 1 and d 2; 5-FU 300 mg/m(2),iv, d 1 and d 2, 5-FU, continuous iv, gtt, 48 h; 1 cycle/2 wk). Response rate, progression-free survival (PFS), total survival time, toxic side effects were evaluated. The inhibitory effect of oxaliplatin on human gastric cell line SGC-7901 was detected and IC(50) was calculated by MTT. Transmission electron microscopy, flow cytometry and TUNEL were performed to evaluate the apoptosis of cell line induced by the drug. The expression of Caspase-3 m-RNA was detected by RT-PCR. AC-DEVD CHO, a Caspase-3 specific inhibitor, was used to elucidate the role of activated Caspase-3 in the process of apoptosis induced by oxaliplatin. RESULTS: Total response (complete and partial) occurred in 9 (40.9%) patients. Mean PFS was 4.2 mo and mean total survival time was 7.2 mo. Cumulative neurotoxicity (all grade I II), vomiting and diarrhea, myelosuppression appeared in 93.5%, 20%, 32.9% patients, respectively. IC(50) was calculated to be 0.71 mg/L by MTT assay. A maximal inhibitory rate reached 85.3%. Apoptosis index was elevated after incubated with 1 mmol/L oxaliplatin for 30 min, but without statistic significance (P>0.05). However it could be detected at a much higher degree both by flowcytometry and by TUNEL with a statistical significance (68.47+/-7.92% and 8.23+/-2.67%, respectively, P<0.05) after incubated with 1 mmol/L oxaliplatin for 2 d. By means of RT-PCR, we detected an enhancement of Caspase-3 m-RNA expression induced by oxaliplatin which was also in positive correlation with the apoptotic level. AC-DEVD-CHO, a Caspase-3 specific inhibitor, could significantly inhibit and delay apoptosis induced by oxaliplatin. CONCLUSION: Oxaliplatin is effective and well-tolerated in patients with advanced gastric carcinoma. Oxaliplatin could significantly inhibit the growth of human gastric cell line SGC-7901. The induction of Caspase-3 m-RNA expression, activation of Caspase-3 and promotion of apoptosis may be some of the therapeutic mechanisms of oxaliplatin on gastric carcinoma. Annexin-V-fluorescein labeling flow cytometry is much more sensitive than TUNEL in detecting early stage apoptosis. PMID- 15334701 TI - A new method of preventing bile duct injury in laparoscopic cholecystectomy. AB - AIM: Of all the complications of laparoscopic cholectecystomy, bile duct injury (BDI) is the most serious complication. The prevention of injury to the common bile duct (CBD) remains a significant concern in laparoscopic cholecystectomy (LC). Different kinds of methods have been advanced to avoid this injury but no single method has gained wide acceptance. Because of various limitations of current methodologies we began a study using cold light illumination of the extrahepatic biliary system (light cholangiography LCP) to better visualize this area and thereby reduce the risk of bile duct injury. METHODS: Thirty-six patients with cholelithiasis were divided into two groups. Group I (16 cases) received LCP and group II (20 cases) received methelenum coeruleum cholangiography (MCCP). In group I cold light was used to illuminate the common bile duct by leading an optical fiber into the common duct with a duodenoscope at the time of LC. The light coming from the fiber in the CBD could clearly illuminate the location of CBD and hepatic duct establishing its location relative to the cystic duct. This method was compared with the dye injection technique using methelenum coeruleum. RESULTS: In group I thirteen cases were successfully illuminated and three failed. The cause of three failed cases was due to the difficulty in inserting the fiber into the ampulla of Vater. No complications occurred in the thirteen successful cases. In each of these successful cases the location of the common and hepatic ducts was clearly seen differentiating the ductal system from surrounding anatomy. In ten cases both the left and right hepatic ducts could be seen and in three only the right hepatic ducts were seen. In four of the thirteen cases, cystic ducts were also seen. In group II, eighteen of the twenty cases were successful. The location of extrahepatic ducts became blue differentiating the ductal system from surrounding anatomy. Two cases failed due to a stone obstructing the cystic duct, and extravisation of the dye turned the entire area blue. LCP showed the common and hepatic ducts more clearly than MCCP. CONCLUSION: LCP is the only technique that can clearly and directly show the location of the extrahepatic biliary system and may be useful in selecting cases of uncertain anatomy in the prevention of bile duct injury. PMID- 15334702 TI - An unusual presentation of "silent" disseminated pancreatic neuroendocrine tumor. AB - To present a patient diagnosed with pancreatic carcinoid that was extremely rare and produced an atypical carcinoid syndrome. We reported a 58-year old male patient who presented with long standing, prominent cervical lymphadenopathy and occasional watery diarrhea. Pathohistological and immunohistochemical examination of lymph node biopsy showed a metastatic neuroendocrine tumor, which was histological type A of carcinoid (EMA+, cytokeratin+, CEA-, NSE+, chromogranin A+, synaptophysin+, insulin-). Bone marrow biopsy showed identical findings. Primary site of the tumor was pancreas and diagnosis was made according to cytological and immunocytochemical analysis of the tumor cells obtained with aspiration biopsy of pancreatic mass (12 mm in diameter) under endoscopic ultrasound guidance. However, serotonin levels in blood and urine samples were normal. It is difficulty to establish the precise diagnosis of a "functionally inactive" pancreatic carcinoid and aspiration biopsy of pancreatic tumor under endoscopic ultrasound guidance can be used as a new potent diagnostic tool. PMID- 15334703 TI - Regulation of plasma erythropoietin in chronic liver disease. PMID- 15334704 TI - Comment on "Eosinophilic gastroenteritis: clinical experience with 15 patients". PMID- 15334705 TI - Organ dysfunction as a risk factor for early severe acute pancreatitis. PMID- 15334706 TI - Importance of a proper placement for your data point. PMID- 15334707 TI - Plagiarized and inaccurate papers in the World Journal of Gastroenterology. PMID- 15334708 TI - Closure of umbilical port in laparoscopic surgery. PMID- 15334709 TI - Laparoscopically assisted versus open colectomy for colon cancer. PMID- 15334710 TI - The physician-supply debate. PMID- 15334711 TI - Donor exchange for renal transplantation. PMID- 15334712 TI - Donor exchange for renal transplantation. PMID- 15334713 TI - A middle ground on public accountability. PMID- 15334715 TI - Polycyclic aromatic hydrocarbons, 15 listings. PMID- 15334716 TI - Procarbazine hydrochloride. PMID- 15334717 TI - Progesterone. PMID- 15334718 TI - 1,3-propane sultone. PMID- 15334719 TI - beta-Propiolactone. PMID- 15334720 TI - Propylene oxide. PMID- 15334721 TI - Propylthiouracil. PMID- 15334722 TI - Radon. PMID- 15334723 TI - Reserpine. PMID- 15334724 TI - Safrole. PMID- 15334725 TI - Selenium sulfide. PMID- 15334726 TI - Silica, crystalline (respirable size). PMID- 15334727 TI - Soots. PMID- 15334728 TI - Streptozotocin. PMID- 15334729 TI - Strong inorganic acid mists containing sulfuric acid. PMID- 15334730 TI - Hospital patient safety incidents account for $6 billion in extra costs annually. PMID- 15334731 TI - Pediatricians need more help with ADHD diagnostic guidelines. PMID- 15334732 TI - Study makes case for using treatment guidelines in early breast cancer. PMID- 15334733 TI - Study says score on stroke test may help streamline care. PMID- 15334734 TI - The future of medical rehabilitation research. PMID- 15334735 TI - Successful pregnancy after combined renal-hepatic transplantation in glycogen storage disease type Ia. AB - Reports of liver and kidney transplantation and of pregnancy in glycogen storage disease type Ia (GSD Ia) (McKusick 232200) are rare. We report an individual who experienced all three successfully. PMID- 15334736 TI - The benefits of liver transplantation in glycogenosis type Ib. AB - There are few reports of liver transplantation in glycogenosis type Ib (GSD Ib). We present two cases who had dramatic catch-up growth and reduced infections after transplantation, despite persistent neutropenia. PMID- 15334737 TI - Early cirrhosis in a patient with type I citrullinaemia (CTLN1). AB - A 17-month-old girl with type I classical citrullinaemia (CTLN1) presenting with early cirrhosis and unusual ultrastructural features of the liver is reported. The patient is homozygous for a splicing mutation in intron 15 of the argininosuccinate synthase gene. PMID- 15334738 TI - Screening for diabetes in general practice. PMID- 15334739 TI - Vitamin E and atherosclerosis. PMID- 15334740 TI - Kelley-Seegmiller syndrome due to a new variant of the hypoxanthine-guanine phosphoribosyltransferase (I136T) encoding gene (HPRT Marseille). AB - A patient with hyperuricaemia and gouty arthritis due to a new variant of hypoxanthine-guanine phosphoribosyltransferase is described. The mutation (I136T, HPRT Marseille) is in the phosphoribosylpyrophosphate-binding region of the gene and leads to almost total loss of enzyme activity in erythrocytes, with 5% in lymphocytes. Nevertheless, the patient showed no neurological abnormality. PMID- 15334741 TI - On: two sessions with Lawrence. PMID- 15334742 TI - On: chaotic possibilities: toward a new model of development. PMID- 15334743 TI - On psychoanalytic education. PMID- 15334744 TI - Rural physicians' acceptance of new Medicare patients. AB - According to the American Academy of Family Physicians (AAFP), the percentage of family physicians accepting new Medicare patients declined from 84.1% in 2000 to 76.1% in 2003 (Trude & Ginsburg 2002). That decline coincided with projected annual decreases in Medicare physician payment announced in March 2002 and March 2003. The decline in the percentage of family physicians accepting new Medicare patients is cause for concern about Medicare beneficiaries' access to health care services. National trends for all physicians may mask different trends among rural physicians. The data in this policy brief describe the trends for urban and rural physicians who no longer accept new Medicare patients. PMID- 15334745 TI - Re: GAMES issue. PMID- 15334746 TI - Re: GAMES issue. PMID- 15334747 TI - Re: GAMES issue. PMID- 15334748 TI - Re: GAMES issue study: Are international genetic consortia functional? PMID- 15334749 TI - [Latest on the epidemiology of headache]. PMID- 15334750 TI - The only constant in Perfusion Education is that it will change. PMID- 15334751 TI - A novel technique of antegrade cerebral perfusion in the newborn with critical aortic stenosis. AB - Various methods of cerebral protection have been used during such aortic arch operations as the Norwood Procedure and operations on the interrupted aortic arch in neonates and infants. Deep hypothermia with circulatory arrest is the most common technique, but has a limited safe period for circulatory arrest. Antegrade cerebral perfusion has been introduced to prolong this safe period. We reviewed our experience with antegrade cerebral perfusion during surgical repair, in a patient with hypoplastic left heart syndrom in stage 1 palliation. PMID- 15334752 TI - Limiting circulatory arrest using regional low flow perfusion. AB - Deep hypothermic circulatory arrest (DHCA) is commonly used for neonatal cardiac surgery. However, prolonged exposure to DHCA is associated with neurologic morbidity. The Norwood operation and aortic arch advancement are procedures that typically require DHCA during surgical correction. Regional low flow perfusion (RLFP) can be used to limit or exclude the use of circulatory arrest. This technique involves cannulation of the innominate or subclavian artery using a Gore-Tex graft, allowing isolated cerebral perfusion. Data was collected in 34 patients undergoing either neonatal aortic arch reconstruction or the Norwood procedure using RLFP. All patients had two arterial pressure monitors using either the umbilical or femoral artery catheters and radial or brachial catheters. Adequacy of perfusion was determined using cerebral saturation, blood flow velocity, mean arterial pressures, and arterial blood gas results. Cerebral saturation and blood flow velocity were monitored using the near-infrared spectroscopy (NIRS) (INVOS 5100, Somanetics Corp, Troy, MI) and a transcranial Doppler pulse-wave ultrasound (TCD) (EME Companion, Nicolet Biomedical, Madison, WI), respectively throughout the entire bypass period. Blood gases were monitored using a point of care blood gas analyzer (Gem Premier, Mallinckrodt Sensor System, Inc., Ann Arbor, MI). Data collected revealed total bypass times for repair between 69-348 min, with a mean of 180 min. Regional low flow perfusion times lasted between 6-158 min, with an average of 50 min., and DHCA times ranged from 0-66 min, with a mean of 19 min. The perfusion techniques used allowed patient clinical data to remain consistent throughout the cardiopulmonary bypass period, regardless of lower flows (Figure 1) The 30-day postoperative mortality rate was 2.9 %, with no evidence of neurologic injury during follow up. In conclusion, regional low flow cerebral perfusion might benefit patients by limiting the use of circulatory arrest during cardiac surgery. Further study is necessary to evaluate patient outcomes, comparing regional cerebral perfusion and circulatory arrest techniques. PMID- 15334753 TI - Comparison of heparin administration using the Rapidpoint Coag and Hepcon HMS. AB - A retrospective chart review was conducted of patients who underwent cardiopulmonary bypass (CPB) to compare the quantities of heparin administered, postoperative blood loss, and homologous blood products transfused during their procedure and subsequent stay in the intensive care unit. The primary purpose of this review was to determine if any difference in heparin administration resulted when two different devices were used for dosing and monitoring heparin. Postoperative blood loss and amount of blood products transfused were also quantified, as any differences would potentially be a result of a difference in administration of heparin. The first group (n = 341) underwent CPB using the Hepcon HMS, Medtronic Inc., Minneapolis, MN, for heparin dosing and monitoring. The Rapid Point Coag, Bayer Healthcare LLC, Tarrytown, NY was used for the second group (n = 345). The two populations were compared for similarity on: age, body surface area (BSA), CPB time (minutes), aortic-cross clamp time (minutes), baseline activated clotting time, and baseline hematocrit. No significant difference was found between the two groups. The second group, using the Rapidpoint Coag, received less heparin during CPB than the group using the Hepcon HMS. In addition, there were decreases in amounts of some blood products transfused as well as mediastinal drainage from the Hepcon HMS to the Rapidpoint Coag group. A summary of the findings can be found in Table 1. A secondary purpose of this study was to determine the influence of hemodilution on the Heparin Management Test (HMT). Citrated whole blood was diluted to varying degrees at various concentrations to determine whether hemodilution with crystalloid would alter the HMT measurements. At all heparin levels and degrees of dilution, the HMT remained stable, with coefficients of variation (CV) of less than 5% at all heparin levels even while incorporating excessive crystalloid dilution (up to 75%). PMID- 15334754 TI - A rapid platelet function assay used to regulate platelet transfusion prophylaxis following cardiopulmonary bypass surgery. AB - Subsequent to cardiac surgery with cardiopulmonary bypass (CPB), patients are at risk of postoperative bleeding caused by acquired defects associated with the procedure. As such, many patients receive prophylactic blood product transfusion. The effectiveness of measuring platelet function using a near-patient platelet function analyzer for the purpose of regulating platelet transfusion and potentially modulating other blood products including fresh frozen plasma (FFP) and cryoprecipitate (CRYO) transfusion was evaluated prospectively in patients undergoing CPB. A 6-month prospective study was designed that encompassed all CPB patients at Aultman Hospital (Canton, OH) during 2001. Each patient was assessed postoperatively for platelet count and function using a point-of-care analyzer. Data were used to help determine whether a platelet transfusion would be administered. Transfusion of CRYO and FFP was also recorded. The data were then compared to the retrospective same 6-month period in the preceding year for the purpose of determining the value of this system in guiding transfusion triage. There were 1770 platelet packs transfused during the study period, representing a 34% reduction in platelet usage. Platelet transfusions were reduced by 51% (5.75 in 2001 vs. 11.75 in 2000) per patient. CRYO/FFP usage was also substantially reduced. During the study period, the patient caseload had increased by 72 (310 vs. 238). Monitoring platelet count and function as a result of CPB and administering an appropriate transfusion protocol can positively impact transfusion outcome. PMID- 15334755 TI - Plateletworks platelet function test compared to the thromboelastograph for prediction of postoperative outcomes. AB - Approximately 3.5 million units of platelets are transfused in the United States each year to patients undergoing open-heart surgery with cardiopulmonary bypass (CPB). CPB is a known contributor to platelet loss and platelet dysfunction leading to disruption of hemostasis. Impaired hemostasis results in excess bleeding in 5-25% of all patients undergoing CPB. For this reason, it may be beneficial to measure platelet number and function in these patients. The purpose of this study was to compare the Plateletworks platelet function analyzer to the thromboelastograph (TEG) in predicting postoperatiave hemostatic outcomes as measured by blood product use and chest tube (CT) drainage. This study consisted of 35 adult patients undergoing cardiac surgery with cardiopulmonary bypass at Rush-Presbyterian-Saint Luke's Medical Center (RPSLMC). The Plateletworks and TEG tests were performed preoperatively, after protamine was given, and 24 hours postoperatively on all patients. Plateletworks demonstrated a statistically significant change in platelet function as shown by the adenosine diphosphate (ADP) reagent tube from the preoperative period to the removal of the aortic cross clamp (p = .011). The TEG did not demonstrate a significant change in the k time and maximum amplitude (MA), but did show a significant change in the alpha angle from the pre-operative to postoperatiave sample (p = .035). A correlation was found between Plateletworks collagen reagent tubes preoperatively and CT drainage (p = .048, r -0.324). No statistical correlation was established between TEG parameters and CT drainage at any time interval. TEG preoperative MA showed a correlation to receipt of blood products (p = .016). When comparing the Plateletworks to the TEG in this study, the Plateletworks system was a more useful predictor of blood product use and chest tube drainage. PMID- 15334756 TI - Observations from a national multiple institution autotransfusion (ATS) quality indicator program. AB - A process to collect universal, mandatory autotransfusion (ATS) procedure quality indicators to measure and monitor ATS quality improvement was designed and implemented by Fresenius Medical Care Extracorporeal Alliance (FMCEA) an outsource provider of extracorporeal services. The indicator program collected and evaluated data that reflect real-world extracorporeal clinical practices and outcomes. The indicator reports provide our clinicians, client physicians, and partner institutions with confidential reports that allow comparison of their practice to evidenced-based performance standards. All ATS procedures reviewed were performed on non-open-heart surgery procedures (on pump or off pump), including vascular, thoracic, orthopedic, and general surgery. After continuous collection and analysis of the indicator data, a hospital is given a report with three components: 1) data analysis that reports summary results and benchmarks the hospital against the other reporting hospitals, 2) corrective action plan that allows the clinical manager to document their investigations and outline plans for continuous quality improvement; and 3) raw data tabulation that allows the clinical manager to identify individual cases that are outliers from the target goal to facilitate local chart reviews. This communication describes FMCEA's ATS Quality Indicator Program and presents the collective results for the first 13 months (January 2002--January 2003) of data collection. Physicians and ATS service client hospitals value the Quality Indicator Process Reports. ATS service managers use the reports and the subsequent process improvement to meet AaBB (American Association of Blood Banks) and JCAHO (Joint Commission on Accreditation of Healthcare Organizations) standards and guidelines for providing safe patient-care services. PMID- 15334757 TI - The combined use of extracorporeal life support and the Berlin Heart pulsatile pediatric ventricular assist device as a bridge to transplant in a toddler. AB - There is a very limited published material about experience with long-term pediatric mechanical circulatory support as a bridge to heart transplant. We report on a 2-year-old, 12 kg boy admitted with 2-week history of low-grade fever, ear pain, pulmonary edema, and congestive heart failure. Trans-thoracic echocardiography confirmed severe myocardial dysfunction with a left ventricular ejection fraction of 0.20 and percentage shortening of 13. After 2 days of ventilatory and inotropic support, the patient continued to deteriorate and subsequently required femoro-femoral extracorporeal life support (ECLS). This was later complicated by a progressive coagulopathy and massive bleeding. On day 17, a pulsatile pediatric paracorporeal biventricular assist device (VAD) (Berlin Heart) was implanted. The patient's condition improved significantly with all coagulopathies corrected, and the patient was extubated 21 days later. After 109 days of bi-VAD support, the patient was successfully transplanted and discharged home 45 days post transplant. Our early experience with initial ECLS bridge to VAD and subsequently to transplant was encouraging. It allowed for additional time to select the ideal organ donor and optimize the recipient's comorbid condition and multiorgan failure. VAD provides an additional armamentarium of circulatory support in pediatric patients with severe heart failure. PMID- 15334758 TI - Evaluation of the Hemobag: a novel ultrafiltration system for circuit salvage. AB - Following termination of bypass, the CPB circuit contains a significant volume of diluted blood. Various methods have been used to salvage this blood, including direct transfusion or centrifugation /washing of the circuit volume. These techniques produce a reinfusion product that is either dilute or free of plasma proteins. The purpose of this study is to evaluate the Hemobag ultrafiltration system, which may overcome these limitations. Yorkshire pigs (n = 4, approximately 40 kg) were placed on CPB (prime volume 1.5 L) for 60 min. Following CPB, control blood samples (Pre) were collected from the circuit. The circuit contents were then transferred into a Hemobag and processed. Blood samples (post) were then collected from the Hemobag. Pre- and post-samples were analyzed and compared using a Student's t-test. Parameters that were significantly different (p < .05) pre-Hemobag versus post-Hemobag were as follows: hematocrit 20.4+/-3.4% vs. 54.1+/-11.6%, total protein 2.4+/-0.4 vs. 8.2+/-2.9 gms/DL, fibrinogen 92.0+/-0.3 vs. 305.8+/-37.2 mg/DL. Parameters that were not significantly different but trended toward an increase post-Hemobag were platelet counts, heparin levels, white cell count, and plasma free hemoglobin. Parameters that showed no differences or trends included sodium, potassium, chloride, bicarbonate, and osmolarity. Processing times were measured at approximately 10 minutes. This device effectively concentrates post-bypass circuit volume, providing a product that is high in red blood cells and plasma proteins and may provide an alternative to current techniques for circuit volume salvage. PMID- 15334759 TI - Aortic valve replacement for a patient with essential thrombocythemia: a case report. AB - A 51-year-old male patient with 3+ aortic insufficiency, hepatitis C, osteomyelitis right foot, and a preoperative platelet count 1.3 million/mm3 required cardiopulmonary bypass (CPB). Essential thrombocythemia is a relatively unknown entity with the utilization of CPB. After consulting with the surgeon, an anesthesiologist and another perfusionist, a team approach was used to discuss treatment for this patient during CPB. The treatment consisted of exchange transfusions, autotransfusion techniques, and current protocol for blood gas management. No incidence of hypercoagulability was observed during this procedure or postoperatively. Based on current knowledge of pathophysiology and review of the literature, plateletpheresis should be the current management of essential thrombocythemia. PMID- 15334760 TI - Mechanical circulatory support: reality and dreams experience of a single center. AB - Because of the increasing number of patients waiting for heart transplantation and the decreasing number of donor organs, mechanical circulatory support has become a generally accepted therapeutic option. Several high-tech devices developed in the last 15 years differ in terms of location, kind of support, and driving units. They are suitable for different patients and their therapeutics objectives. Based on 13 years of experience, we developed a specific protocol for selection and management of patients and devices. Six hundred two patients have received mechanical circulatory support (MCS) in our institution since 1987. The indication spectrum includes cardiogenic shock for various reasons: acute myocarditis, right heart failure, acute rejection and postcardiotomy heart failure, alternative to transplantation, and bridge to recovery. Eight different systems are in use at our center. The extracorporeal devices, the Biomedicus centrifugal pump (n = 169) and the Abiomed BVS 5000 (n = 92) are used for short term support. The Thoratec VAD (n = 179), and Medos HIA-VAD (n = 10) located in paracorporeal position preferably used for midterm support. Novacor LVAS (n= 96), and HeartMate (n = 58) are partially implantable systems used for long-term ventricular assistance in patients who did not require biventricular support. The advantage of the implantable devices is the option of discharging patients under support if they fulfill special criteria before being discharged to home. Eighty five LVAD patients were discharged home with support, Novacor (n = 52), HeartMate (n = 27), ThoratecTLC-II (n = 8), Lionheart (n = 3) fulfill our criteria for being discharged home while on support. Careful postoperative patient management does not exclude a variety of complications. Bleeding: occurred in 22-35% of patients, right heart failure in 15-26%, neurologic disorder in 7-28%, infection in 7-30%, and liver failure in 11-20%. Complications varied with different devices, and the patients' preoperative conditions. Eighty-five patients fulfilled the criteria of our out of hospital program (OOH) and were discharged from hospital for a mean period of 184 days. Readmission was necessary for complications caused by thromboembolism and infection. This report describes our patient device selection criteria as a bridge to transplant setting. PMID- 15334761 TI - Plasma-modified ACT can be used to monitor bivalirudin (Angiomax) anticoagulation for on-pump cardiopulmonary bypass surgery in a patient with heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a problem of growing significance and concern, affecting an estimated 1-3% of patients among those receiving an estimated 10 million heparin exposures annually in the United States. The prevalence of heparin antibodies has been reported as at least 12.7% in the general cardiac surgery population before surgery and 42% following surgery. Various management techniques for anticoagulation in these patients have been proposed. Many of these alternative agents present additional risks for bleeding and efficacy and, furthermore, require the use of nonconventional monitoring assays to assess the level of anticoagulation achieved adequately. We report here the successful use of bivalirudin anticoagulation on pump, with no additional morbidity, and the first reported use of the plasma-modified ACT (activated clotting time) test, a simple modification of the standard ACT assay, to monitor the anticoagulant effect of bivalirudin. PMID- 15334762 TI - Argatroban usage for anticoagulation for ECMO on a post-cardiac patient with heparin-induced thrombocytopenia. AB - We report a post-Norwood Stage I patient requiring ECMO support using Argatroban as an anticoagulant following diagnosis of heparin-induced thrombocytopenia (HIT). A 2.6 kg female was born with hypoplastic left heart syndrome and underwent a Norwood Stage I operation on day 4 of life. The patient weaned off cardiopulmonary bypass with no complications and was routinely placed on a ventricular assist device (VAD) for 3 days. Heparin was infused at a rate of 16 32 IU/ kg/h to maintain an ACT of 160-180 seconds. Two days after VAD termination, the patient was placed on continuous veno-veno hemofiltration (CVVH). Shortly after CVVH, the patient was diagnosed with HIT and placed on an Argatroban infusion. Five days later, a VAD and subsequent ECMO was used because of decreasing left ventricular function, gross body edema, and poor renal function. This case report summarizes the use of Argatroban during VAD and ECMO support for a patient diagnosed with HIT. PMID- 15334763 TI - The use of cardiopulmonary bypass and extracorporeal membrane oxygenation for support during removal of two teratomas and hydrops fetalis. AB - The removal of massive tissue tumors often leads to rapid blood loss and decreased lung compliance because of large volume shifts. Cardiopulmonary bypass (CPB) and extracorporeal membrane oxygenation (ECMO) have both been used as a means of support during resection of saccrococcygeal teratomas. Hydrops fetalis is the accumulation of fluid in extravascular spaces and body cavities. This leads to edema and sometimes hypoxia. ECMO has been used for support during treatment of hydrops fetalis. This patient was diagnosed, via sonogram, at 30 weeks gestation to have two teratomas and hydrops fetalis. Because of the risk of hemorrhage and poor lung compliance during removal of these types of tumors, CPB and ECMO were used. This support allowed for successful removal of the tumors. Following removal of the tumors, the patient failed to wean from CPB because of the severity of hydrops. The patient remained on extracorporeal support for treatment of hydrops fetalis. This report describes the perfusion techniques used for support during the removal of teratomas and the treatment of hydrops fetalis. PMID- 15334764 TI - The effects of leukocyte reduction on matrix metalloproteinase release in cardiopulmonary bypass. AB - Matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading the extracellular matrix, a process that likely contributes to the development of altered vascular permeability. Past studies in patients undergoing cardiopulmonary bypass (CPB) have documented increased levels of MMPs with CPB. The purpose of this study was to evaluate the effect of leukocyte reduction on MMP release during CPB. Patients (n = 17) undergoing elective coronary revascularization requiring CPB were randomly assigned to either a leukocyte reducing filter (LRF) group (n = 9) or the standard CPB circuit with no LRF (n = 8). White blood cell (WBC) counts, MMP-2, and MMP-9 levels were serially measured at baseline and up to 12 hours post CPB. MMP levels were measured by enzyme linked immunoassay. ProMMP-2 levels increased in both the non-LRF and LRF groups but to a higher degree in the LRF group. ProMMP-9 levels increased by 40% in the non-LRF group. In contrast, proMMP-9 decreased by 30% in the LRF group. The addition of leukocyte-reducing filters in the CPB circuit attenuated the release of MMP-9 but increased release of MMP-2 post-CPB. Because MMPs can degrade the extracellular matrix, leading to increased vascular permeability, attenuation of MMPs may have decreased the local tissue injury known to occur as a result of these enzymes. However, future prospective studies to test this hypothesis directly are warranted. PMID- 15334765 TI - Rationale of platelet gel to augment adaptive remodeling of the injured heart. AB - Cardiac pathologic events including; myocardial infarction, viral infection and hypertrophy, and aging, may trigger maladaptive remodeling of the myocardium. Maladaptive remodeling results in diastolic and systolic dysfunction, myocyte loss, and malformation of the extracellular matrix. It is proposed that platelet gel applied to the site of myocardial injury may provide the proper cytokines, growth factors, and chemokines to promote adaptive remodeling. The hypothesis is that platelet gel concentrates may provide a temporary and local hyperphysiologic concentration of platelet secretory factors that may initiate adaptive myocardial healing. Autologous platelet gel can be derived from concentrated platelets activated and induced to secrete cytokines, growth factors, and chemokines with an array of stimulating agents. This report discusses selected platelet secretory factors, including; IL-1beta, TGF-beta, TGF-alpha, FGF, EGF, PDGF, and IGF, which support the concept that platelet concentrates can mediate cardiac wound healing. In conclusion, application of platelet gel to areas of cardiac injury may offer a therapeutic means to stimulate myocyte regeneration, angiogenesis, and restoration of a normal extracellular matrix composition. PMID- 15334766 TI - Relationship of aging and cardiac IL-10. AB - Current therapies for the treatment of myocardial infarction and heart failure include medical, surgical, mechanical assist, and transplantation. These therapies have been based on the dogma that ventricular myocytes themselves are terminally differentiated and, therefore, cannot regenerate. This concept has been recently challenged with stem cell therapy. A potential problem is the ability of cardiac tissue to mobilize, recruit, and transdifferentiate adult stem cells from other tissues. We believe that there is a unique failure of the damaged myocardium to provide the appropriate molecular signals for stem cells engraftment related to age. Our hypothesis is that the overexpression of IL-10 in the aged population reduces cardiac cellular proliferation subsequent to myocardial injury. This hypothesis is supported by aging models, where elevated levels of IL-10 are associated with reduced healing response to noncardiac tissue injury. We demonstrated an increased cardiac gene expression of IL-10 that may be associated with a reduced proliferative response in the border regions of the infarcted myocardium that are proportional with age. In conclusion, myocardial infarction and heart failure has presented a significant challenge for the clinician to provide reparative therapies. The use of therapeutics to modulate IL 10 and, thereby, optimizing regenerative processes in the injured myocardium may provide a unique means for the cardiac patient. PMID- 15334767 TI - Classic pages of the Journal of ExtraCorporeal Technology. PMID- 15334768 TI - The aetiology and clinical relevance of gastric intestinal metaplasia. PMID- 15334769 TI - Duodenal alkaline secretion: its mechanisms and role in mucosal protection against gastric acid. AB - Duodenal mucosa, especially its proximal portion, is exposed to intermittent pulses of gastric acid (H+). This review summarises the mechanisms of duodenal bicarbonate (HCO3-) secretion and their role in protecting duodenal epithelium against gastric H+. Duodenal epithelium is a leaky barrier against gastric H+, which diffuses into duodenocytes, but fails to damage them due to: (a) an enhanced expression of cyclooxygenase, producing protective prostaglandins and expression of nitric oxide synthase, releasing nitric oxide, both stimulating duodenal HCO3- secretion and (b) the release of several neurotransmitters also stimulating HCO3- secretion such as vasoactive intestinal peptide, pituitary adenylate cyclase-activating polypeptide, acetylcholine and melatonin. At the apical duodenocyte membrane, several HCO3-/Cl- anion exchangers operate in response to luminal H+ to extrude HCO3- into duodenal lumen. In baso-lateral duodenocyte membrane, both non-electrogenic and electrogenic Na+-HCO3- cotransporters are activated after exposure of duodenum to gastric H+, causing inward movement of HCO3- from extracellular fluid to duodenocytes. There are also at least three Na+/H+ exchangers, eliminating H+ which diffused into these cells. The Helicobacter pylori infection with gastric metaplasia in the duodenum and bacterium inoculation results in the inhibition of HCO3- secretion by its endogenous inhibitor dimethyl arginine, resulting in ulcerogenesis. PMID- 15334770 TI - Frontal cortical perfusion abnormalities related to gluten intake and associated autoimmune disease in adult coeliac disease: 99mTc-ECD brain SPECT study. AB - OBJECTIVE: Since brain perfusion abnormalities have been described by single photon emission computed tomography in some autoimmune diseases, the aim of the present study was to evaluate the incidence of perfusion abnormalities by brain single-photon emission computed tomography in a group of coeliac disease patients, and to investigate whether gluten intake and associated autoimmune diseases may be considered risk factors in causing cerebral impairment. METHODS: Thirty-four adult coeliac patients (16 on a gluten-free diet and 18 on a gluten containing diet, 18 (53%) with autoimmune diseases) underwent 99mTc-ethyl cysteinate dimer brain single-photon emission computed tomography and qualitative evaluation of brain perfusion was performed together with a semiquantitative estimation using the asymmetry index. Ten subjects on our database, matched for sex, age and ethnic group, who were proved normal by histology ofjejunal mucosa (four males and six females; median age 39 years, range 27-55 years), were included as control group. RESULTS: Twenty-four out of 34 patients (71%) showed brain single-photon emission computed tomography abnormalities confirmed by abnormal regional asymmetry index (>5%; range 5.8-18.5%). Topographic comparison of the brain areas showed that the more significant abnormalities were localised in frontal regions, and were significantly different from controls only in coeliac disease patients on unrestricted diet. The prevalence of single-photon emission computed tomography abnormalities was similar in coeliac disease patients with (74%) and without (69%) associated autoimmune disease. CONCLUSIONS: Abnormalities of brain perfusion seem common in coeliac disease. This phenomenon is similar to that previously described in other autoimmune diseases, but does not appear to be related to associated autoimmunity and, at least in the frontal region, may be improved by a gluten-free diet. PMID- 15334771 TI - Wireless capsule video endoscopy compared to barium follow-through and computerised tomography in patients with suspected Crohn's disease--final report. AB - BACKGROUND: Wireless capsule endoscopy is a superior diagnostic tool to barium small bowel follow-through and enteroscopy in diagnosing patients with occult blood loss. AIM: To compare capsule endoscopy with barium follow-through and entero-computerised tomography in patients with suspected Crohn's disease. SUBJECTS AND METHODS: Thirty-five patients with suspected Crohn's disease underwent the three examinations. The radiologist and gastroenterologist were blinded to each other's results. In cases of discrepancy, colonoscopy and ileoscopy were performed. RESULTS: Thirty-five patients (22 males), mean age 28.4 years, were included. Eighty-eight percent had abdominal pain, 83% had diarrhoea and 69% had weight loss. The diagnostic yield of capsule endoscopy was 77% versus 23% and 20% of barium and computerised tomography examinations, respectively (P < 0.05). The capsule detected all of the lesions diagnosed by barium follow-through and entero-computerised tomography. CONCLUSIONS: Capsule endoscopy is a superior and more sensitive diagnostic tool than barium follow-through and entero computerised tomography in patients with suspected Crohn's disease. PMID- 15334772 TI - Costs associated with outpatient diarrhoea in infants and toddlers: a nationwide study of the Italian Society of Paediatric Gastroenterology and Hepatology (SIGEP). AB - BACKGROUND: As diarrhoea mortality is negligible in Italy, other costs should be considered when planning health strategies. Little is known about the costs associated with diarrhoea in Italian children. AIMS: To assess the costs associated with outpatient infantile diarrhoea in Italy. METHODS: Primary care paediatricians from five regions filled in a questionnaire for the first 10 children (1-47 months) they visited for acute diarrhoea during a 3-month period. RESULTS: We analysed 473 questionnaires. Mean age (standard deviation) of children was 21 (11) months; mean duration of diarrhoea (standard deviation) was 4.3 (2.6) days. An overall cost of 110 (137) euro per episode was estimated, with significant difference between children younger and older than 36 months (116 euro versus 72 euro). Missed work by relatives accounts for 75% of the cost. The parents of children attending a day-care centre had an increased risk to miss work (relative risk = 2.15). A weak relationship was found between days of diarrhoea and missed work (r = 0.30); it could be estimated that the diarrhoea should be shortened by about 4 days in order to save I day of missed work. CONCLUSIONS: Acute outpatient diarrhoea is associated with a significant financial burden in Italy. Simply shortening the diarrhoea does not seem to be the most expeditious way to reduce the cost of diarrhoea itself. PMID- 15334773 TI - Prevalence of the K469E polymorphism of intercellular adhesion molecule 1 gene in Italian patients with inflammatory bowel disease. AB - BACKGROUND: Intercellular adhesion molecule 1 plays an important role in the recruitment of leucocytes at sites of inflammation and is up-regulated in intestinal mucosa of inflammatory bowel disease. Intercellular adhesion molecule 1 gene lies on chromosome 19p13, implicated in determining susceptibility to inflammatory bowel disease. Recently, the polymorphism K469E of intercellular adhesion molecule 1 gene has been identified. AIM: To assess the potential association of this polymorphism with inflammatory bowel disease. PATIENTS: A total of 165 inflammatory bowel disease patients, 75 with Crohn's disease and 90 with ulcerative colitis, and 187 controls were studied. METHODS: The K469E polymorphism was detected by polymerase chain reaction and restriction enzyme analysis. Statistical analysis was performed by chi2-test. RESULTS: In inflammatory bowel disease, the distribution of intercellular adhesion molecule 1 genotypes was 24.9% E/E, 44.2% E/K and 30.9% K/K. In controls, 11.8% showed E/E genotype, 55.6% E/K and 32.6% K/K. The frequency of the E/E genotype was significantly higher in inflammatory bowel disease (Crohn's disease and ulcerative colitis) patients than in controls. Subgroup analysis showed that the frequency of the E469 allele was significantly increased only in Crohn's disease patients with ileocolonic location of disease and penetrating behaviour compared with controls. CONCLUSIONS: We found an association of inflammatory bowel disease with the E/E genotype of intercellular adhesion molecule 1 gene, while allele E469 was associated with a subgroup of Crohn's disease patients with more extensive location of disease and penetrating behaviour. However, further studies are needed to confirm our findings. PMID- 15334774 TI - Quality-related variables at hepatological websites. AB - BACKGROUND: The amount of hepatology-related information available on the Internet has substantially increased, but little is known about the characteristics and quality of the websites. AIM: The aim of this study was to describe analytically and evaluate critically the information concerning three diseases of hepatological interest: chronic hepatitis, hemochromatosis and Caroli's disease. METHODS: In accordance with a validated method, the three search terms were entered into four English language search engines and the first five links of each were considered (a total of 60 sites). The characteristics of the websites were described and their quality was evaluated by three independent reviewers who assigned scores of 1-5 for accuracy, reliability and depth. The relationships between the site characteristics and quality scores were analysed by means of multiple logistic regression. RESULTS: The overall rating score was sufficient (>3) in 51% (95% confidence interval: 38-65%) of cases. The majority of the sites (73%) were aimed at patients rather than at physicians. Commercial sponsorship was significantly more frequent among the chronic hepatitis sites (45%) than among the hemochromatosis (15%) or Caroli's disease sites (0%) (P = 0.002); 61% of the commercial sites did not include a financial disclosure. The only variable that independently related to poor quality was the presence of commercial sponsorship (odds ratio 18.1; 95% confidence interval: 1.7-192.5). CONCLUSIONS: Hepatological websites are characterised by poor quality and are mainly aimed at patients. Quality is negatively affected by commercial interests, which are often undeclared. Guidelines for the certification and surveillance of websites relating to liver diseases are highly advisable. PMID- 15334775 TI - The rational use of albumin in patients with cirrhosis and ascites. A Delphi study for the attainment of a consensus on prescribing standards. AB - BACKGROUND: Ascites is one of the most frequent severe complications in patients with liver cirrhosis. The treatment of this chronic disease usually requires the prolonged use of albumin, frequently continued even after patients' discharge from the hospital. AIMS: Aim of the study was to define a consensus among Italian physicians with regard to the use of albumin in patients with decompensated cirrhosis and ascites. METHODS: The study adopted the Delphi technique to conduct the consensus activities. All controversial issues related to the use of albumin were identified by the experts' board and proposed to the 68 participating hepatology centres through two subsequent questionnaires. The questionnaires, returned by the specialists involved, were collected and the answers classified to verify the elements on which a consensus was reached. RESULTS: The home use of albumin can help to improve the patient's general conditions and well-being. About 77% of the experts involved considered likely that albumin administration could shorten hospital stays or could reduce the number of hospital admissions. The results of the study, along with a socioeconomic analysis, were presented to the Italian Drug Commission, which subsequently removed the specific hypoalbuminemia level as a prerequisite for having the drug reimbursed by the National Health Service. CONCLUSIONS: For an outpatient prescription, the hypoalbuminemia limit of 2.5 g/dl or less is not sufficient, while the decision whether to administer the drug requires the evaluation of patient's overall clinical conditions as an essential criterion for the prescription of a home treatment with albumin. PMID- 15334776 TI - Reported risk factors are useless in detecting HCV-positive subjects in the general population. AB - BACKGROUND: Chronic hepatitis C virus infection is mostly asymptomatic, so it will not be identified if specific diagnostic tests are not performed. AIMS: To evaluate the positive predictive value of some risk factors in detecting anti hepatitis C virus-positive subjects in the general population. SUBJECTS: Two thousand five hundred and sixty-one subjects randomly selected from the list of the census in three population-based surveys performed in hepatitis C virus endemic areas in Southern Italy. METHODS: The sensitivity, specificity and positive predictive value of blood transfusion, past use of glass syringes and surgical intervention in detecting hepatitis C virus positivity were assessed. Data were collected using a precoded questionnaire administered by an interviewer. RESULTS: All risk factors showed a poor positive predictive value (ranging from 21.0% for surgical intervention to 29.0% for blood transfusion). The positive predictive value was extremely low (ranging from 2.9 to 4.3%) in subjects younger than 46 years of age, who mostly could benefit from antiviral treatment. The combination of the simultaneous presence of more than one risk factor does not improve the detection of hepatitis C virus infection. CONCLUSIONS: Reported risk factors are useless in detecting hepatitis C virus positive subjects in the general population. PMID- 15334777 TI - Mesalazine-induced reversible infertility in a young male. AB - Mesalazine is a well-established treatment for ulcerative colitis. A young man treated with mesalazine for proctitis was found to have pathological semen with a count of only 3 x 10(6) sperm cells/ml. He failed to achieve conception with his wife. When mesalazine treatment was stopped, semen analysis returned to near normal and pregnancy ensued. Due to recurrent disease activity, mesalazine treatment was reinstituted. This was followed by deterioration of the patient's semen. PMID- 15334778 TI - Unusual cystic presentation of an endocrine carcinoma of the jejunum. AB - The cystic presentation of endocrine tumours is rare and raises difficult diagnostic problems. So far, the only cases of cystic digestive endocrine tumours reported in the literature are of pancreatic origin. We report the unusual observation of a jejunal endocrine carcinoma presenting as a cystic abdominal mass. A 59-year-old woman was referred for chest and abdominal pain. Imaging studies revealed multiple cystic nodules in the liver and a large sus-mesocolic cystic lesion of probable intestinal origin. Biopsies of the extra-hepatic mass and liver nodules showed endocrine tumour. Surgical resection of the jejunal mass and of liver segment III were performed. Histological examination confirmed the diagnosis of jejunal endocrine carcinoma metastatic to the liver. Large areas of the primary and secondary tumours presented an unusual vesicular architecture, responsible for the cystic presentation. No adjuvant treatment was attempted. This observation underlines the difficult diagnostic problems raised by the cystic presentation of digestive endocrine tumours. PMID- 15334779 TI - Recurrent intussusception associated with Escherichia coli 0157 infection. PMID- 15334780 TI - Infliximab and quality of life in steroid-dependent Crohn's disease. PMID- 15334781 TI - Distal duodenal surveillance in familial adenomatous polyposis. PMID- 15334782 TI - [Sense and nonsense about aging. Discrimination of the elderly, generational differences and perspectives]. PMID- 15334783 TI - [Is good suicide possible?]. PMID- 15334784 TI - [Determinants of care burden among spouses and children of physically impaired elderly]. AB - The study focused on determinants of care burden among spouses and children providing care to older adults. The care recipients in this study are older adults who participated in the Longitudinal Aging Study Amsterdam (LASA) since 1992. A selected subsample of 155 chronically ill respondents with a need for care, 78 of their spouses and 337 of their children participated in a side-study on family care giving. The results show that 32% of the spouses and 40% of the children assists the older adult with personal and/or domestic care. These spouses have provided care for many years with both domestic as well as personal care. The care providing children predominantly assist with household activities and share these tasks with siblings. Children are more likely to provide care when the spouse is not available or not able to provide care. Professional care does not effect children's care behavior. Half of the spouse-carers and one-fifth of the child-carers experienced heavy care burden. A higher care burden is associated with providing personal care, experiencing more negative consequences, and a larger need for assistance. Burdened child-carers also report lower feelings of competence, less consulting with siblings and more disagreement with siblings on caregiving issues. These findings point out that caregiving is not an individual task but a family matter. Support of carers should not only be directed at a decrease of the care load and an strengthening of individual capacities, but also at recruiting other informal (family) caregivers in order to divide the load of care giving more evenly among all those involved. PMID- 15334785 TI - [The diagnostic value of drawing a clock in geriatrics]. AB - In this study the Clock Drawing Test (CDT) is used in a geriatric department to search for dementia in a population with somatic disorders (N=236). The demented group (N=113) has significant lower scores on every item and on the total score of the scoring method, compared to the non-demented group (N=123). A factor analysis of the scoring method shows that the fourteen items can be grouped into the three groups "contour", "digits", and "hands". A logistic regression shows that "hands" is the most discriminating, although this effect is moderate (sensitivity 59.3%, specificity 75.6%). Considering however that the CDT, compared to other screening tasks, appeals to an unique combination of cognitive functions, including visual semantic memory, working memory, visuospatial skills, attention and executive functions, it can be concluded that this test can contribute to the diagnoses of dementia, besides the traditionally used instruments. PMID- 15334786 TI - [Detection of dementia of the Alzheimer type with the 7 Minutes Test (7 MT)]. PMID- 15334787 TI - [Ischemic colitis]. AB - Ischaemic colitis is the most frequent form of mesenteric ischaemia and is most frequently the result of a non-occlusive disease process. Elderly patients, especially those with underlying co-morbid conditions are at greatest risk. The symptoms are often non-specific and diagnosis is usually established upon the clinical setting and colonoscopy with biopsy. In the majority of cases, ischaemic colitis can be treated non-surgically with a successful outcome. PMID- 15334788 TI - [Use it and lose it]. PMID- 15334789 TI - Diabetes mellitus decreases hippocampal release of neurotransmitters: an in vivo microdialysis study of awake, freely moving rats. AB - Possible diabetes mellitus-induced changes in hippocampal monoaminergic activities were studied to understand the relationships between neurotransmitter levels and various abnormalities in freely moving diabetic rats. We used both experimentally (STZ rats) and spontaneously diabetic rats (WBN/Kob rats) as the diabetic animal model, and compared the findings with those obtained from non diabetic rats (C rats). Measurement of neurotransmitters (serotonin and dopamine) was carried out using an in vivo microdialysis method. We found that: 1) the basal level of serotonin in the hippocampus was lowest in WBN rats, followed by STZ rats, then by C rats. The level of serotonin in WBN rats was about a half of that in C rats; 2) the basal level of dopamine was also significantly lower in the diabetic WBN and STZ rats than in C rats. The data show that diabetes mellitus decreases in the monoamine release from the hippocampus in both experimentally and spontaneously diabetic rats. PMID- 15334790 TI - Modification of beta-cell response to different postprandial blood glucose concentrations by prandial repaglinide and combined acarbose/repaglinide application. AB - This study was designed to compare the effects of repaglinide plus acarbose combination treatment to repaglinide alone on postprandial glucose, serum insulin, C-peptide and proinsulin concentrations. A total of 40 patients with Type 2 diabetes (T2DM) (fasting blood glucose: 120-180 mg/dl; postprandial blood glucose: 140-240 mg/dl) were included in this single-centre, controlled, randomised, single-dose, cross-over study. On two consecutive days, patients either received 2 mg repaglinide 15 min before breakfast followed by 100 mg acarbose with breakfast or repaglinide alone. Two fasting (7.30 h, 8.00 h) and five postprandial blood samples (from 8.30 h to 12.00 h) were taken for blood glucose, serum insulin, C-peptide and proinsulin determination. Repaglinide plus acarbose treatment significantly reduced the mean increase in postprandial blood glucose levels (24.2+/-18.2 mg/dl) compared to repaglinide alone (51.1+/-29.0 mg/dl; p<0.001). Serum insulin, C-peptide and proinsulin levels [mean area under the curve (AUC7.30-12.00h)] were significantly lower than those observed with repaglinide monotherapy (e.g. insulin: 1089.2+/-604.5 hr x pmol/l and 1596.8+/ 1080.6 hr x pmol/l, resp., p<0.001), suggesting that acarbose modifies the rapid insulin release induced by repaglinide. Prandial treatment with a combination of acarbose and repaglinide results in an additive glucose lowering effect and modified insulin secretion compared to repaglinide alone. Postprandial hyperglycaemia is not abolished by rapid stimulation of insulin release induced by repaglinide. Additional reduction of postprandial blood glucose by acarbose modifies the stimulation of insulin release. PMID- 15334791 TI - Metabolic variations with oral antidiabetic drugs in patients with Type 2 diabetes: comparison between glimepiride and metformin. AB - Patients with Type 2 diabetes (T2DM) are at high risk of morbidity and mortality from cardiovascular complications, and hypoglycaemia increases this risk. Furthermore, other metabolic parameters exacerbate cardiovascular risk in these patients. The aim of the study was to compare the metabolic effects of glimepiride and metformin in patients with T2DM. We evaluated 164 patients with T2DM (80 males, 84 females) in a multicentre, randomised, controlled, open, parallel group study comparing glimepiride with metformin. Eighty-one patients (aged 56+/-10 yr) received glimepiride (3+/-1 mg/d); 83 patients (aged 58+/-9 yr) received metformin (2500+/-500 mg/d). Patients had been diagnosed for < or = 6 months; they were non-smokers; had no hypertension or coronary heart disease; were not taking hypolipidaemic drugs, diuretics, beta-blockers or thyroxin; and had normal renal function. Metabolic parameters were measured after 6 and 12 months of treatment. Glimepiride significantly lowered lipoprotein(a) [Lp(a)] and homocysteine levels (HCT) at 6 and 12 months. Both glimepiride and metformin lowered plasminogen activator inhibitor Type 1 (PAI-1) at 12 months and significantly improved levels of glycosylated haemoglobin, fasting plasma glucose and post-prandial plasma glucose after 6 and 12 months. Metformin significantly lowered fasting plasma insulin and postprandial plasma insulin. Glimepiride and metformin also reduced levels of other metabolic parameters in patients with T2DM. In particular, glimepiride significantly reduced HCT, Lp(a), and PAI-1 levels, important metabolic risk factors for atherosclerotic vascular disease. These reductions may be owing to improved glucose metabolism, but it cannot be excluded that these drugs have a direct effect on additional metabolic parameters. PMID- 15334792 TI - Quality of life and coping behaviour in Type 1 diabetes mellitus: relationship with metabolic control. AB - Uncovering factors possibly leading to insufficient metabolic control in Type 1 diabetes, both on the part of the patient or the treating physician, is of considerable relevance. The present long-term study investigated the relevance of patient-related vs education-related factors for the success in achieving acceptable glycaemic control. Adolescents or young adults with Type 1 diabetes mellitus (n= 26, mean age= 22+/-2 yr, diabetes duration= 11+/-5 yr) were followed during 36+/-5 months. All patients were treated by the same diabetologist. At the beginning of the study coping behaviour, quality of life and evaluation of emotional status were assessed. Changes in HbA1c were used as a parameter of glycaemic control. At follow-up there was a significant decrease in HbA1c of 0.4% (p<0.01). However, this was not in statistically significant correlation with age, gender, aspects of quality of life or coping behaviour. Therefore, glycaemic control and/or improvement of glycaemic control in adolescents or young adults with Type 1 diabetes mellitus seems to be primarily related to other factors, eg continuous education provided in a stable setting. PMID- 15334793 TI - Antibiotic therapy for diabetic foot infections: comparison of cephalosporines with chinolones. AB - Diabetic foot complications are the most common cause of non-traumatic lower extremity amputations and uncontrolled infections represent a major risk factor. This open prospective, multicenter trial compared the efficacy of two antibiotic regimens for treatment of foot infections Wagner stage II or III in diabetic adults. Three hundred diabetic patients with severe, limb-threatening foot infection were consecutively enrolled in a prospective, observational, matched pairs controlled study to test two different antibiotic regimes (ceftriaxone vs chinolones) in addition to standard treatment of foot infection. After matching, 90 patients--each receiving ceftriaxone or chinolones--were analyzed. Our study demonstrated that treatment with a third generation cephalosporine is as effective as a treatment with chinolones. Response (reaching Wagner I or 0) was achieved in 58.0% in the ceftriaxone group and in 51.1% in the chinolone group (NS.). Fourteen days after initiation of treatment, the number of patients with microbiological isolates decreased in both groups (52 to 5 in the ceftriaxone group and 60 to 12 in the chinolone group). At hospital discharge, 66.0% of ceftriaxone and 64.4 of chinolone-treated diabetic ulcers were cured or improved. In summary, both substances proved to be effective in the primary antibiotic treatment of the diabetic foot; an early broad spectrum antibiotic treatment, that covers both gram-positive and gram negative bacteria as well as anerobes is undisputedly an imperative therapeutic intervention for the treatment of diabetic foot infection. PMID- 15334794 TI - Impact of low frequency transcutaneous electrical nerve stimulation on symptomatic diabetic neuropathy using the new Salutaris device. AB - In a double blind, randomised study, 19 patients suffering from mild-to-moderate symptomatic diabetic neuropathy (Total Symptom Score, NTSS 4-16) received either treatment with the new transcutaneous electrical nerve stimulation (TENS) device "Salutaris" (verum group) or a placebo treatment with an identical but electrically inactive device (placebo group). Stimulation pads were placed at the anatomical localisation of the peroneal nerve and stimulation was performed using a low frequency mode. At baseline (V1), after 6 (V2), and 12 (V3) wk of treatment, the patients' symptoms were registered using the new total symptom score (NTSS-6) and a visual analogue scale (VAS). In addition, sensory nerve thresholds (temperature, vibration, pain) and microvascular function were measured at the lower limb at baseline and after 12 wk of treatment. Active TENS treatment resulted in a significant improvement in NTSS-6 score after 6 wk (-42%) and after 12 wk (-32%) of treatment (baseline: 10.0+/-3.3, 6 wk: 5.8+/-5.0, p<0.05; 12 wk: 6.8+/-3.9, p=0.05; placebo group: baseline: 7.6+/-3.1; 6 wk: 8.1+/ 5.1, n.s.; 12 wk: 6.5+/-6.1, n.s.). Subanalysis of the different qualities of the NTSS-score revealed an improvement in numbness (2.2+/-1.0 to 1.6+/-1.3; p<0.03); lancinating pain (1.6+/-1.1 to 0.6+/-0.9; p<0.02) and allodynia (1.4+/-1.6 to 0.5+/-1.0; p<0.05). Also, a significant improvement in the VAS rating was found after 6 wk of TENS therapy (19.8+/-5.0 to 14.4+/-9.6; p<0.05), while no change was observed in the placebo arm. In conclusion, our study indicates that the new TENS device "Salutaris" is a convenient, non-pharmacological option for primary or adjuvant treatment of painful diabetic neuropathy. PMID- 15334795 TI - Is it time to use the new lower limit of impaired fasting glucose (IFG) among the ATP III criteria for diagnosis of the metabolic syndrome? Prevalence rate of the metabolic syndrome using old and new lower limits of IFG in obese and/or hypertensive subjects. PMID- 15334796 TI - Urinary stone disease in Sri Lanka. PMID- 15334797 TI - Genetically modified food: friend or foe? PMID- 15334798 TI - Chloroquine resistant falciparum malaria among security forces personnel in the Northern Province of Sri Lanka. AB - OBJECTIVE: To determine the occurrence and species distribution of malaria and the extent of chloroquine resistance among security forces personnel in a selected region of the Northern Province of Sri Lanka. DESIGN: A descriptive study. SETTING: Mannar District in the Northern Province. METHODS: Nine hundred and seventy five security personnel were screened for malaria by microscopy. Those who were positive were treated with chloroquine and were subjected to 28 day in vivo assay to determine chloroquine resistance. In vitro microtest assay was performed to determine the response of Plasmodium falciparum isolates to chloroquine in vitro. RESULTS: Of the 975 personnel screened, 181 (18.6%) were positive for malaria. P. falciparum was the predominant species (n = 125; 69.1%). The rest were due to P. vivax (n = 42; 23.2%) and mixed infections (n = 14; 7.7%). This was an inversion of the usual species distribution pattern in the country. In vivo assay revealed 38 (53.5%) P. falciparum infections as chloroquine resistant. Fifteen of 23 (65.2%) P. falciparum isolates showed evidence of resistance in vitro. None of the P. vivax infections showed evidence of chloroquine resistance. There was no significant difference in the severity of clinical disease between chloroquine resistant and sensitive infections at first presentation. Recrudescent P. falciparum infections had significantly lower mean parasite densities as well as lower clinical scores at recrudescence than at first presentation. CONCLUSION: Results demonstrate the high prevalence of malaria and chloroquine resistance in the study area and explains several contributory factors for this. There is an urgent need to review antimalarial drug policies in Sri Lanka. PMID- 15334799 TI - An audit of state sector intensive care services in Sri Lanka. AB - OBJECTIVE: To survey the facilities, functioning characteristics, bed strength, manpower, operational practices and the distribution of the Intensive Care Units (ICU) of Government Hospitals in Sri Lanka. DESIGN: A cross-sectional observational study. METHOD: Interview of the sister or the nurse in charge of each unit by telephone using a structured questionnaire. SETTING: Department of Anaesthesiology, Faculty of Medicine, University of Peradeniya. STUDY POPULATION: All intensive care units of the government hospitals in Sri Lanka. MEASUREMENTS: Bed strength, facilities, functioning characteristics, manpower and equipment. RESULTS: Fifty two intensive care units were identified in the island. Two units could not be contacted over the telephone and one refused to participate. Of the 49 ICUs studied 28 (57.1%) were located in teaching hospitals, six (12.2%) in provincial hospitals, 13 (26.5%) in base hospitals and two (4.1%) in special hospitals. Twenty five (51%) of the 49 ICUs were multidisciplinary, three (6.1%) general medical, four (8.2%) general surgical and the remainder were of medical, surgical and paediatric subspecialities. The minimum acceptable standard of a ventilator: bed ratio of 1:1 was seen in 28 (57%) and a nurse : bed ratio of 1:1 was seen in 37 (75.5%) ICUs. A 24-hour resident medical officer was available in 46 (93.9%) of the 49 ICUs. ICUs are mostly located in larger cities. The lowest ICU coverage (one ICU for about 1.2 million people) was seen in the Uva Province. CONCLUSIONS: ICUs in Sri Lanka are mainly located in teaching hospitals. The standards and management strategies vary widely. PMID- 15334800 TI - Couple characteristics and outcome of therapy in vaginismus. AB - OBJECTIVE: To describe couple characteristics and outcome of therapy in vaginismus. DESIGN: A prospective before-after intervention descriptive study. SETTING: Department of Physiology, Faculty of Medicine, Colombo, Sri Lanka. METHODS: Fifty six couples with vaginismus, mostly self referrals and referrals from gynaecologists and general practitioners, were treated with a standard cognitive behaviour therapy protocol with before-after assessments of the degree of vaginismus and individual partner self-ratings of the relationship and psychological status (GHQ-30). OUTCOME MEASURES: Success at the end of the therapy was equated to the absence of or only mild vaginismus and, improvement in the couple relationship and psychological status scores. RESULTS: Twenty seven (48%) of the 52 (93%) couples with non-consummation reported failure of coitus following previous non-surgical and surgical interventions. Love marriages (70%), frequent attempts at sex (75%, 3 or 4 times/week) and sexual arousal (women = 86%, men = 89%) characterised couples. Ten men developed sexual problems, mostly erectile failure and premature ejaculation secondarily. Couple therapy enabled penetrative sex in 45 (80.3%). The single prognostic indicator of outcome was the degree of vaginismus at first visit, those with mild and moderate vaginismus (77%) being significantly more likely to establish coitus (p<0.001) and complete the therapy (p<0.001). The couple relationship improved marginally (women p<0.01, men 0.025

0.01) but the psychological status remained unchanged. Dropouts and referrals for psychiatric and marital counselling failed to complete therapy. INTERPRETATION: Couple sex therapy is effective in the management of vaginismus. Health professionals, especially gynaecologists and general practitioners, need to be aware of the problem and the satisfactory outcome of sex therapy. PMID- 15334801 TI - Chryseobacterium meningosepticum infections in a dialysis unit. AB - BACKGROUND: Chryseobacterium species are Gram-negative bacteria with an unusual antibiotic profile. Chryseobacterium meningosepticum is the species most commonly encountered as a human pathogen. OBJECTIVES: To study the microbiological, clinical and therapeutic features of C. meningosepticum infections in patients on dialysis, at Sri Jayewardenepura General Hospital (Teaching) (SJGH), and to trace the source of infections. DESIGN: A retrospective descriptive study. SETTING: Dialysis unit of SJGH. PATIENT: population Patients who underwent long term haemodialysis (HD) and manual intermittent peritoneal dialysis (IPD) in the dialysis unit. METHODS: Clinical and microbiological records of patients with C. meningosepticum infections over a period of 2 years were reviewed retrospectively. Environmental screening was carried out to detect a possible source of infection. RESULTS: Thirty five episodes of infection due to C. meningosepticum in 33 patients on HD and IPD were detected. There were 30 episodes of peritonitis, four of bacteraemia and one of asymptomatic colonization of a PD catheter. Isolates were resistant to aminoglycosides, chephalosporins and aztreonam, and sensitive to cotrimoxazole, vancomycin and rifampicin. They showed variable sensitivity to imipenem and ciprofloxacin. All except one patient had a favourable outcome. C. meningosepticum was cultured from a sink in the dialysis unit, but the original source of the organism was not known. CONCLUSION: C. meningosepticum could be an important pathogen in a dialysis unit, and fluoroquinolones and vancomycin are effective as empiric therapy. PMID- 15334802 TI - Blood lead levels of children before and after introduction of unleaded petrol. PMID- 15334803 TI - Assessment of a child mental health inpatient unit. PMID- 15334804 TI - Bertiella studeri infection: resistance to niclosamide. PMID- 15334805 TI - Giant hydronephrosis: a rare cause of gross abdominal distension. PMID- 15334806 TI - The outcome of elevated prostate specific antigen and transrectal ultrasound guided prostatic biopsy in detecting carcinoma of the prostate: initial experience in Sri Lanka. PMID- 15334807 TI - Prevalence and control of risk factors in patients after acute coronary events. PMID- 15334808 TI - Unacceptable teacher behaviour or medical student abuse? PMID- 15334809 TI - Do schools promote violence in Sri Lanka? PMID- 15334810 TI - Detection and management of alcohol misuse by general practitioners. PMID- 15334811 TI - Cross-national comparisons of social isolation and loneliness: introduction and overview. AB - This article provides an introduction to the special volume on research related to issues of social isolation and loneliness for older people. It discusses some key concepts and provides a brief overview of the articles included in the volume. PMID- 15334812 TI - Changes in levels of social isolation and loneliness among older people in a rural area: a twenty-year longitudinal study. AB - The Bangor Longitudinal Study of Ageing (BLSA), conducted in rural Wales from 1979 to 1999, followed a cohort of survivors from more than 500 people over 20 years. Using both quantitative and qualitative data from the study, the factors associated with increases and decreases in loneliness and social isolation were identified. The study was based on a population sample and survivors were followed up every 4 years. From 1983 to 1987, 30 people aged 75 and over in 1979 were studied intensively. The customary measure of loneliness was used, as well as an aggregate measure devised by the research team. Social isolation was similarly measured, using an aggregate measure. Respondents were assessed as demonstrating low, moderate, or high levels of loneliness or isolation. Subsequently, statistical models of loneliness and social isolation were developed. Some respondents were assessed as not experiencing social isolation or loneliness during the study. Others showed changes in levels. In this article, the data are explored, seeking factors associated with changes in social isolation and loneliness. Outcome measures of these two variables of interest are compared with items from the aggregate measures and other identifiable intervening variables. The article discusses which change variables contribute most to levels of isolation and loneliness and result in different combinations of these two outcomes. Implications for policy and practice are discussed. PMID- 15334813 TI - Social isolation and loneliness: differences between older rural and urban Manitobans. AB - The purpose of this paper was to identify predictors of social isolation and loneliness for very old rural and urban adults. With data from the 1996 Aging in Manitoba Study (N = 1,868; age range 72-104), separate multiple regression models were constructed for rural and urban sub-samples, using the life space index (LSI) to measure social isolation as one outcome, and a loneliness index created by the authors from a combination of items to measure loneliness as a second outcome. Different factors were found to predict the outcomes for the two sub samples. The models with isolation as the outcome produced five predictors for the rural sub-sample and three for the urban sub-sample. Only living alone was the same for both groups. The models with loneliness as the outcome produced five predictors for the rural sub-sample and two for the urban sub-sample, again with only one factor in common--four or more chronic illnesses. We conclude that health and social factors are important predictors of social isolation and loneliness, and sensitivity to these factors may improve the experience of older adults. PMID- 15334814 TI - Gender and marital-history differences in emotional and social loneliness among Dutch older adults. AB - In this study, Weiss's (1973) theorizing about the sources of emotional and social loneliness is elaborated--with notions about the asymmetric gratifications derived from marriage, about the conflicting loyalties that result from remarriage, and about selection into marriage--in order to reach an understanding of gender differences in loneliness, both in and outside of marriage. First and subsequent marriages are considered, as well as marital disruptions and never marrying. The data (N = 3737) are from the 1992 Dutch survey on older adults' living arrangements and social networks (NESTOR-LSN). Marital-history differences emerge, not only for emotional loneliness, but also (and contrary to Weiss's theoretical conceptualizations) for social loneliness. The marital-history differences in emotional and social loneliness are greater among men than women. For men, the marriage bond appears not only to be more central to emotional well being than is the case for women but also to play a pivotal role in their involvement with others. Marital history offers the best explanation for differences in emotional loneliness among men, but social embeddedness characteristics also account for differences in emotional loneliness among women. Apparently, whereas men are more likely to find an intimate attachment in marriage, women also find protection from emotional loneliness in other close ties. The marital-history differences in social loneliness are largely mediated by social embeddedness characteristics, partly in different ways for men and women. Involvement in activities outside the home serves as the context for sociability for men, whereas parenthood plays a more important role in women's social engagements. PMID- 15334815 TI - Old age and loneliness: cross-sectional and longitudinal analyses in the Tampere Longitudinal Study on Aging. AB - The purpose of this study was to examine whether older age is associated with increasing loneliness in people aged 60 and over. Data came from TamELSA, a population-based prospective longitudinal study in Tampere, Finland. The follow up time was 20 years. Loneliness was measured by a single question--"Do you feel lonely?"--with the possible answers often, sometimes, or never. Cross-sectional analysis showed that the percentage of subjects feeling lonely increased toward older age groups, but in a multivariate analysis, only household composition and social participation were independently associated with loneliness. Longitudinal analysis showed that loneliness increased with higher age. Over a 10-year period, loneliness increased most in those who, at baseline, were married and living alone with their spouse. In conclusion, only a minority of older people continuously suffer from loneliness. Loneliness does increase with age, not because of age per se, but because of increasing disability and decreasing social integration. PMID- 15334816 TI - Loneliness among older adults in The Netherlands, Italy, and Canada: a multifaceted comparison. AB - Loneliness is experienced in many cultures. To properly assess cross-cultural differences, attention should be paid to the level, determinants, and measurement of loneliness. However, cross-cultural studies have rarely taken into account more than one of these. Differences in the level of loneliness were hypothesized on the basis of national differences in partnership, kinship, and friendship, which were assumed to be related to cultural standards within a society. Differences were examined among married and widowed older adults aged 70 to 89 years living independently in The Netherlands (N = 1,847), Tuscany, Italy (N = 562), and Manitoba, Canada (N = 1,134). Loneliness was measured with an 11-item scale. The Manitobans were high on emotional loneliness and the Tuscans were high on social loneliness. Partner status excepted, the determinants were nearly the same across the three locations. Differential item functioning (DIF) related to the three locations was observed for most items. Interactions with gender and the availability of a partner relationship were observed. PMID- 15334817 TI - European and Canadian studies of loneliness among seniors. AB - This article provides a commentary on a set of five other articles reporting European and Canadian studies of loneliness among seniors. It places those works involving Canadian, Dutch, Finnish, and Welsh samples in the larger context of research on loneliness; offers reflections on the methods and findings reported in the articles; and addresses the question, Is loneliness universal? Points of similarity in the articles are identified and possible ways of reconciling discrepant findings regarding age trends and gender differences are put forward. A discrepancy model of loneliness is used as a key framework for explaining several points, including why objective social isolation and loneliness don't always go together. PMID- 15334818 TI - [Evolutionary transformations of ontogenesis in anuran amphibians]. AB - A review of the recent published data on ontogenesis of direct developing and marsupial frogs. The development of these representatives of anuran amphibians seems to be evolutionary advanced and considerably differs from the development of species traditionally used in amphibian embryology. PMID- 15334819 TI - [Age-related features of ceruloplasmin biosynthesis and distribution in rats]. AB - Biosynthesis of ceruloplasmin, a copper-containing glycoprotein, which plays an important role in copper transfer and bidirectional iron transport in vertebrates, was studied in 7-day old rats characterized by the embryonic type of copper metabolism. In addition to the liver, copper is synthesized in the lungs, brain, and kidneys. The appearance of labeled ceruloplasmin in the blood flow coincides with the time of liberation of de novo synthesized ceruloplasmin from the liver. [14C]-Ceruloplasmin is absorbed from the blood flow by cells of the heart, lung, and kidneys and binds to erythrocytes. The polypeptide ceruloplasmin chain does not enter the brain cells from the blood flow. Immunoreactive polypeptides of the ceruloplasmin receptor were found using immunoblotting in plasma membranes of the heart, liver, kidneys, and erythrocytes, rather than in those of the brain. It was shown by reverse transcription coupled with PCR using selective primers these cells contain molecular forms of ceruloplasmin mRNAs programming the synthesis of both secretory ceruloplasmin and ceruloplasmin connected with the plasma membrane via the glycosyl phosphatidylinositol anchor. After transition to the adult type of copper metabolism, the blood serum contents of copper and ceruloplasmin sharply increase, while the content of CP in the cerebrospinal fluid, as measured according to the oxidase and antigen activities, and copper concentration, as determined by atom-absorption spectrometry, remain low. Ontogenetic features of the system ensuring the copper homeostasis in mammals are discussed. PMID- 15334820 TI - [Peripheral pool of CD8+ T-cells contains lymphocytes with antigen-specific receptors that recognize syngeneic MHC class II molecules]. AB - The capacity of T-lymphocytes to recognize "nonself" and tolerating "self" is formed as a result of positive and negative selection in the thymus. While obtaining and testing specificity of T-hybridomas, we demonstrated that the major part of peripheral pool of CD8+ T-lymphocytes carried receptors specific to "self" MHC class II molecules. Such an unexpected specificity of receptors has been found in some T-cell hybridomas produced by fusion of activated peripheral CD8+ T-lymphocytes with a tumor partner transfected by the coreceptor CD4 gene. The reactivity to "self" is not an experimental artifact due to an increased avidity of interaction of the hybridoma cells with antigen-presenting cells. Also, it is not an expression of reactivity of T-cells to superantigens, products of endogenous viruses of mouse breast cancer. The formation of a pool of such T cells involves both cells with double receptor specificity and cells coexpressing two alpha-chains of T-cell receptor. Their appearance in the periphery can be due to the capacity of thymocytes differentiating in the direction of CD4+ cells to avoid negative selection via change of expression of coreceptor CD4 to CD8. PMID- 15334821 TI - [Structurization of cortical layer of loach yolk cell after wounding as a "minimal" model of morphogenesis]. AB - Structural rearrangements of the yolk cell surface were studied in loach embryos using SEM and TEM, which take place within 30 min after a point-like puncture at the late blastula stage. The effects of sucking off or addition of a part of yolk, lowered temperature, and absence of Ca2+ on structurization were studied. Around the area of puncture, the yolk granules were submerged, the number of vesicles increased, and numerous membrane folds were formed. The folds were aggregated to form two sharply distinct types of structures: a group of rounded evaginations around the site of puncture and a system of radial folds in the periphery. Small radial folds are aggregated in radial strands, several dozens folds in each. Sucking off a part of yolk accelerated the above processes, while addition of yolk, cooling, and absence of Ca2+ in the incubation medium slowed down or suppressed these processes. The observed structurization can be considered as self-organization at the level of the yolk cell cortical level, largely similar to that during normal morphogenesis at the level of multicellular sheets. Hence, the membrane dynamics in the yolk cell wall after its damage can be considered as one of simplified ("minimal") models of morphogenesis. A study of this model makes it possible to narrow down the circle of factors essential for self-organization of morphogenetic processes. PMID- 15334822 TI - [Development of loach eggs after experimental increase of cell mass in the dorsal and ventral areas of blastoderm]. AB - The fertilized loach eggs were injected, before the beginning of cleavage, with the nuclear dye Hoechst 33258 and left to develop until the late blastula stage. Some cells of the dorsal area of stained blastoderm were transplanted in the analogous area of intact embryos of the same age, which led to an earlier and more pronounced development of head and trunk structures in recipients. A relationship was established between specific features of the development of recipients and localization of descendants of the transplanted cells. Transplantation of cells of the dorsal area of stained blastoderm in the ventral area of embryos of the same age led to the formation of two axial complexes, both at the same level of development, nut behind the control, and stained cells were located predominantly in one of twin embryos. PMID- 15334823 TI - [Effect of prenatal stress on the pituitary-adrenal axis in blue foxes]. AB - Handling is a source of stress for farm bred blue foxes. The influence of handling during the late gestation period on the pituitary--adrenal axis was studied in 10-day old male and female blue foxes. Cortisol and progesterone were measured by radioimmunoassay in the plasma, adrenal homogenates, and in vitro incubates from animals of both sexes. Adrenals were incubated in vitro in the absence or presence of ACTH. In addition, the adrenal weight and plasma concentration of ACTH were assessed. In cubs of both sexes, the adrenal weight was decreased after prenatal stress. The plasma concentration of progesterone and the adrenal cortisol in vitro production were elevated in the prenatally stressed female cubs, as compared to the control, along with the adrenal progesterone in vitro production in prenatally stressed male cubs. The adrenal cortisol and progesterone content and plasma ACTH and cortisol concentrations were not affected by prenatal stress. In conclusion, the results of this study suggest that the prenatal stress induced by handling pregnant vixens can affect the pituitary--adrenal axis in neonatal offspring, this effect being more pronounced in female cubs. PMID- 15334824 TI - [Effect of of methoprene and retinoic acid on characteristics of Haller's organ regeneration in the tick Haemaphysalis longicornis (Acari, Ixodidae)]. AB - The results of experiments on regeneration of the Haller's sensory organ in the metastriate ixodid tick Haemaphysalis longicornis Neumann in the presence of the juvenoid methoprene and retinoic acid and of studies of its structural changes by SEM confirmed the similarity of prostriate and metastriate ixodids as concerns the juvenalizing effect of the above morphogens on regenerative processes during nymphal-imaginal metamorphosis. However, the metastriate ticks (Haemaphysalis and earlier studied Hyalomma) are well behind the prostriate ticks (Ixodes) as concerns the extent of changes induced by juvenoids and retinoids in the sensillar sets of Haller's organ regenerates. PMID- 15334825 TI - [Saving insertional recessive lethal mutants of Arabidopsis thaliana]. AB - A group of 13 recessive lethal mutants was selected on the basis of the collection of Arabidopsis thaliana transgenic plants with insertions of T-DNA vector plasmid pLD3 or pPCVRN4, which was produced by agrobacterial transformation of germinating seeds. The use of media containing exogenous hormones made it possible to compensate the lethal effect, identify phenotypes, and characterize six lines of recessive lethal germlings using genetic and molecular-genetic methods. PMID- 15334826 TI - [Comparative characteristics of immunological, clinical and morphological features of human lymphoid tumors in respect with tumor genesis and progression]. AB - We describe three lymphoid tumors with the same immunophenotype characteristic for chronic lymphoid leukemia (CD19+/CD5+, clonality of the light immunoglobulin chains, CD23+ and CD10-). However, clinical picture and morphology of neoplastic cells dictate different clinical forms of these cases: chronic lymphoid leukemia, large cell transformation of chronic lymphoid leukemia and diffuse large B-cell lymphoma. Taking into account that immunophenotype reflects the origin of tumor, while clinical outcome and morphological features of cells reflect the stage of tumor progression and/or pathway of tumor formation, we discuss the approach to natural classification of lymphoid tumors based on the process of their evolution. PMID- 15334827 TI - The role of PK/PD parameters to avoid selection and increase of resistance: mutant prevention concentration. AB - The continuing escalation of antimicrobial resistant human pathogens and the limited number of new antimicrobial agents under development has dictated that our knowledge on the emergence of resistance and any potential strategies to slow the rate at which resistance occurs is of paramount importance. Investigations with fluoroquinolones resulted in the mutant prevention concentration (MPC) concept which represents a novel in vitro measurement of fluoroquinolone potency. In essence, the MPC defines the antimicrobial drug concentration threshold that would require an organism to simultaneously possess two resistance mutations for growth in the presence of the drug. An alternative definition is the drug concentration that prevents the growth of first-step resistant mutants or the minimal inhibitory concentration of the most resistant organism present in the heterogeneous bacterial population when tested against > or =10(9) organisms. From in vitro investigations, the new fluoroquinolones (gatifloxacin, gemifloxacin, moxifloxacin) were all found to have lower MPC values than did levofloxacin against clinical isolates of Streptococcus pneumoniae. Ciprofloxacin was found to have lower MPC values than levofloxacin against clinical isolates of Pseudomonas aeruginosa. When MPC data is applied to achievable and sustainable serum drug concentrations in the body, estimation of the time the serum drug concentration exceed both MIC and MPC values can be determined. This data along with kill data allows for an estimate of the amount of time drug concentration needs to exceed MIC/MPC values to not only result in significant kill but also to minimize resistance development. To date, MPC measurements have been determined in in vitro microbiological and pharmacological models and animal and human data are being investigated. The data summarized in this overview detail resistance issues for P. aeruginosa, S. pneumoniae and other pathogens. Also presented is a summary of the MPC concept and investigations completed to date. A brief summary of fluoroquinolone mechanisms of action and resistance is presented. Finally, some preliminary investigations with other classes of compounds are discussed. To date, very limited data is available to conclude if the MPC concept does or does not apply to other classes of antimicrobial agents. PMID- 15334828 TI - Supraspinal connections of the reproductive organs: structural and functional aspects. AB - Gonadal functions are governed by the hypothalamo-hypophyseal system. Other organs of the reproduction tract are under the regulatory action of gonadal steroids. In the past two decades several data have been accumulated on the involvement of fine-tuning control mechanisms which include autocrine and paracrine effects of biologically active substances produced locally and the regulatory action of nerves innervating the organs of the system. Recent studies using the viral transsynaptic technique have revealed cell groups in the central nervous system that are transneuronally connected with the male and female reproductive organs. This review summarizes neuromorphological data on the supraspinal innervation of reproductive organs and the functional significance of these brain areas in the control of reproduction. PMID- 15334829 TI - Different cardiorespiratory responses to hemorrhage and hyperoxia in normotensive (WKY) and spontaneously hypertensive (SHR) rats. AB - OBJECTIVE: To compare the cardiorespiratory responses underlying the beneficial effects of hyperoxia during blood loss between normotensive (WKY) and hypertensive (SHR) rats. METHODS: Experiments were carried out in anesthetized animals with both carotid bifurcations either innervated or denervated. The effects of breathing 60% O2 in N2 were studied either in combination with non hypotensive hemorrhage or during hemorrhagic hypotension. RESULTS: In normoxia arterial pressure fell more in SHR than in WKY for a given blood loss. During hyperoxia, nerve-intact rats showed initial suppression of ventilation, but bifurcation-denervated rats a powerful enhancement. In all groups, hyperoxia increased the overall tone of venous capacitance vessels. CONCLUSIONS: The greater blood loss in SHR than in WKY when bleeding down to a given arterial pressure results from a stronger constriction of venous capacitance vessels. Hyperoxia improves the ability of the cardiorespiratory system to resist the effects of hemorrhage by increasing the overall venous tone, thus supporting cardiac filling, and in some cases also by increasing alveolar ventilation, probably secondary to improved cerebral oxygenation. The beneficial effects of hyperoxia were: (i) not prevented by carotid denervation, and thus were presumably direct tissue effects of oxygen, (ii) strikingly weaker in SHR than in normotensive (WKY) rats. PMID- 15334830 TI - Morphological and functional differences in cardiac parameters between power and endurance athletes: a magnetic resonance imaging study. AB - The present study compared morphological and functional parameters of the left ventricle by magnetic resonance imaging (MRI) in competitive athletes engaged in endurance and power activities and sedentary control subjects. Twenty male subjects, 7 endurance-trained athletes (ETA) (age 23.8+/-3.5 yr), 7 strength trained athletes (STA) (age 22.8+/-4.0 yr), and 6 sedentary controls (age 24.1+/ 2.2 yr) were studied by MRI. In the ETA group body size related left ventricular mass (rel.LVM) was significantly higher than that in the STA group (71.0+/-9.2 vs 57.4+/-15.7 g/m3). The difference between their size related left ventricular wall thickness (rel.LVWT) values (9.37+/-1.0 vs 8.37+/-1.8 mm/m) was near to the level of significance (p=0.057). Relative left ventricular internal diameter (rel.LVID) was significantly higher in the ETA group compared to the STA group (42.3+/-1.0 vs 40.1+/-2.5 mm/m, p<0.05). The muscular quotient (MQ=LVWT/LVID) of the ETA group was not significantly higher compared to the strength athletes. Relative left ventricular end-diastolic volume (LVEDV) was also higher in the ETA group than in the STA group (69.5+/-6.7 vs 59.9+/-8.2 ml/m3, p<0.05) and the controls (53.6+/-3.7, p<0.001). Significantly higher relative stroke volume (SV) was measured in the ETA group compared to the STA group and the controls (41.0+/ 5.7; 32.6+/-6.9; 32.0+/-3.2 ml/m3). According to the present data, the strongest impact on LV cavity size and wall thickness is caused by long-term high intensity endurance training. Intense strength training does not necessarily induce wall thickening. PMID- 15334831 TI - Decreased chemotaxis of human peripheral phagocytes exposed to a strong static magnetic field. AB - The chemotaxis of human peripheral phagocytes, neutrophils and monocytes was examined in a strong static magnetic field (0.317+/-0.012 Tesla). The chemotaxis of the suspension of purified neutrophils and monocytes was tested in the Boyden chamber using C5a as a chemotactic signal. The chambers were placed into a temperature regulated (36.6 degrees C) equipment producing a strong static magnetic field (0.317 Tesla) for 60 minutes. The movement of cells proceeded into a nitrocellulose membrane toward the north-pole of the magnet, i.e. in the direction of the Earth's gravitational pull. The C5a induced chemotaxis of human neutrophils decreased significantly in the strong static magnetic field. Monocytes were not significantly effected. The strong static magnetic field decreased the chemotactic movement of neutrophils and this phenomenon may have implications when humans are exposed to magnetic resonance imaging for extended periods of time. PMID- 15334832 TI - Relation between pharmacodynamic and anthropometric parameters during ergometry at rest and after repeated intake of metoprolol in healthy volunteers: results of a pilot study. AB - Metoprolol is a widely used beta1-selective beta-blocker in hypertension and tachycardia. The influence on vital signs at rest and during ergometry (exercise heart rate or blood pressure, effect areas above baseline) was investigated in a pilot study with 18 healthy volunteers (mean age 29.1 years) by means of multiple and pairwise correlation analysis. At rest, the difference between predose and day 5 values were not associated with anthropometric characteristics. During ergometry for weight and height significant negative correlations were found corresponding to marked beta-values in the multiple regression models. Therefore heart rate decreases less markedly in slim persons which should be taken into consideration in exercise tests during metoprolol intake. PMID- 15334833 TI - Effect of orally administered plant lectins on intestinal liquor accumulation and amylase activity in rats. AB - Short-term effects of orally administered plant lectins, with special reference to the Phaseolus vulgaris agglutinin (phytohaemagglutinin, PHA), were studied in growing rats. The orally administered PHA elicited a dose-dependent accumulation of liquor with elevated pH in the proximal small intestine. Although the concentration of alpha-amylase activity did not change, total alpha-amylase activity slightly, but significantly increased in the gut. When a panel of plant lectins with different carbohydrate binding specificities was tested at the dose of 100 mg/kg body weight, most of them stimulated the secretion of liquor, but the total alpha-amylase activity was increased only by PHA, ConA or WGA. PMID- 15334835 TI - Effect of omeprazole paste on intragastric pH in clinically normal neonatal foals. AB - OBJECTIVE: To evaluate the efficacy of omeprazole paste, a commonly used antiulcer drug, on intragastric pH in clinically normal neonatal foals. ANIMALS: 6 clinically normal foals between 5 and 14 days of age. PROCEDURE: Intragastric pH was recorded in each foal by use of a disposable antimony pH electrode with internal reference. Values for intragastric pH were recorded every 4 seconds by use of an ambulatory pH monitor. There were two 24-hour recordings of intragastric pH for each foal, with 24 hours between recordings. Foals were not administered any drugs during the first recording. Foals were administered omeprazole paste (4 mg/kg, PO) 1 hour after the start of the second recording. Mean pH was calculated for each hour of each 24-hour recording session. Hourly mean values were compared between the first and second 24-hour recordings. RESULTS: Complete data were obtained from 4 of 6 foals during the first 24-hour recording and 6 of 6 foals during the second 24-hour recording. Foals had significantly higher mean hourly intragastric pH for hours 2 to 22 following omeprazole administration, compared with corresponding hourly pH values in foals during the first recording. CONCLUSION AND CLINICAL RELEVANCE: Omeprazole paste can effectively increase intragastric pH in clinically normal neonatal foals within 2 hours after oral administration of the first dose and can be administered to neonatal foals at the rate of 4 mg/kg, PO, every 24 hours. PMID- 15334836 TI - Intracranial elastance in isoflurane-anesthetized horses. AB - OBJECTIVE: To determine whether high intracranial pressure (ICP) during spontaneous ventilation (SV) in anesthetized horses coincides with an increase in intracranial elastance (ie, change in ICP per unit change of intracranial volume). ANIMALS: 6 adult horses. PROCEDURE: Anesthesia was induced and maintained in each horse for 5 hours with isoflurane at a constant dose equal to 1.2 times the minimum alveolar concentration. Direct ICP measurements were obtained by use of a strain gauge transducer inserted in the subarachnoid space, and arterial blood pressure was measured from a carotid artery. Physiologic responses were recorded after 15 minutes of normocapnic controlled ventilation (CV) and then after 10 minutes of SV. Aliquots (3 mL) of CSF were removed from each horse during SV until ICP returned to CV values. Slopes of pressure-volume curves yielded intracranial elastance. RESULTS: Intracranial elastance ranged from 0.2 to 3.7 mm Hg/mL after removal of the first aliquot of CSF Slopes of pressure-volume curves were largest following removal of the initial CSF aliquot, but shallow portions of curves were detected at relatively high ICPs (25 to 35 mm Hg). A second-order relationship between SV ICP and initial intracranial elastance was found. CONCLUSIONS AND CLINICAL RELEVANCE: In horses anesthetized with isoflurane, small changes in intracranial volume can cause large changes in ICP Increased intracranial elastance could further exacerbate preexisting intracranial hypertension. However, removal of small volumes of CSF may cause rapid compensatory replacement from other intracranial compartments, which suggests steady-state maintenance of an increase in intracranial volume during isoflurane anesthesia in horses. PMID- 15334837 TI - Risk of postnatal exposure to Sarcocystis neurona and Neospora hughesi in horses. AB - OBJECTIVE: To estimate risk of exposure and age at first exposure to Sarcocystis neurona and Neospora hughesi and time to maternal antibody decay in foals. ANIMALS: 484 Thoroughbred and Warmblood foals from 4 farms in California. PROCEDURE: Serum was collected before and after colostrum ingestion and at 3 month intervals thereafter. Samples were tested by use of the indirect fluorescent antibody test; cutoff titers were > or = 40 and > or = 160 for S neurona and N hughesi, respectively. RESULTS: Risk of exposure to S neurona and N hughesi during the study were 8.2% and 3.1%, respectively. Annual rate of exposure was 3.1% for S neurona and 1.7% for N hughesi. There was a significant difference in the risk of exposure to S neurona among farms but not in the risk of exposure to N hughesi. Median age at first exposure was 1.2 years for S neurona and 0.8 years for N hughesi. Highest prevalence of antibodies against S neurona and N hughesi was 6% and 2.1 %, respectively, at a mean age of 1.7 and 1.4 years, respectively. Median time to maternal antibody decay was 96 days for S neurona and 91 days for N hughesi. There were no clinical cases of equine protozoal myeloenchaphlitis (EPM). CONCLUSIONS AND CLINICAL RELEVANCE: Exposure to S neurona and N hughesi was low in foals between birth and 2.5 years of age. Maternally acquired antibodies may cause false-positive results for 3 or 4 months after birth, and EPM was a rare clinical disease in horses < or = 2.5 years of age. PMID- 15334838 TI - Comparison of the effects of caudal pole hemi-meniscectomy and complete medial meniscectomy in the canine stifle joint. AB - OBJECTIVE: To compare the effects of caudal pole hemi-meniscectomy (CPHM) and complete medial meniscectomy (MM), specifically with respect to development of secondary osteoarthritis, in the stifle joints of clinically normal dogs. ANIMALS: 14 large-breed dogs. PROCEDURE: Unilateral CPHM (7 dogs) or MM (7) was performed, and the left stifle joints served as untreated control joints. Gait was assessed in all dogs before surgery and at 4, 8, 12, and 16 weeks postoperatively. After euthanasia, joints were evaluated grossly; Mankin cartilage scores, subchondral bone density assessment, and articular cartilage proteoglycan extraction and western blot analyses of 3B3(-) and 7D4 epitopes were performed. RESULTS: Weight distribution on control limbs exceeded that of treated limbs at 4 and 16 weeks after surgery in the CPHM group and at 4 and 8 weeks after surgery in the MM group; weight distribution was not significantly different between the 2 groups. After 16 weeks, incomplete meniscal regeneration and cartilage fibrillation on the medial aspect of the tibial plateau and medial femoral condyle were detected in treated joints in both groups. Mankin cartilage scores, subchondral bone density, and immunoexpression of 3B3(-) or 7D4 in articular cartilage in CPHM- or MM-treated joints were similar; 7D4 epitope concentration in synovial fluid was significantly greater in the MM-treated joints than in CPHM-treated joints. CONCLUSIONS AND CLINICAL RELEVANCE: Overall severity of secondary osteoarthritis induced by CPHM and MM was similar. Investigation of 7D4 epitope concentration in synovial fluid suggested that CPHM was associated with less disruption of chondrocyte metabolism. PMID- 15334839 TI - Evaluation of microbial culture of pooled fecal samples for detection of Mycobacterium avium subsp paratuberculosis in large dairy herds. AB - OBJECTIVE: To evaluate sensitivity of microbial culture of pooled fecal samples for detection of Mycobacterium avium subsp paratuberculosis (MAP) in large dairy herds and assess the use of the method for estimation of MAP prevalence. ANIMALS: 1,740 lactating cows from 29 dairy herds in California. PROCEDURE: Serum from each cow was tested by use of a commercial ELISA kit. Individual fecal samples were cultured and used to create pooled fecal samples (10 randomly selected fecal samples/pool; 6 pooled samples/herd). Sensitivity of MAP detection was compared between Herrold's egg yolk (HEY) agar and a new liquid culture method. Bayesian methods were used to estimate true prevalence of MAP-infected cows and herd sensitivity. RESULTS: Estimated sensitivity for pooled fecal samples among all herds was 0.69 (25 culture-positive pools/36 pools that were MAP positive). Sensitivity increased as the number of culture-positive samples in a pool increased. The HEY agar method detected more infected cows than the liquid culture method but had lower sensitivity for pooled fecal samples. Prevalence of MAP-infected cows was estimated to be 4% (95% probability interval, 2% to 6%) on the basis of culture of pooled fecal samples. Herd-level sensitivity estimate ranged from 90% to 100% and was dependent on prevalence in the population and the sensitivity for culture of pooled fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: Use of pooled fecal samples from 10 cows was a cost-effective tool for herd screening and may provide a good estimate of the percentage of MAP-infected cows in dairy herds with a low prevalence of MAP. PMID- 15334840 TI - Plasma mineral and energy metabolite concentrations in dairy cows fed an anionic prepartum diet that did or did not have retained fetal membranes after parturition. AB - OBJECTIVE: To compare plasma total calcium, phosphorus, magnesium, nonesterified fatty acids (NEFA), beta hydroxy butyrate (BHB), and glucose concentrations in parturient dairy cows that were fed an anionic prepartum diet between those with and without retained fetal membranes (RFM) at 24 hours after parturition. ANIMALS: 152 Holstein cows that calved during October through December of 1997 PROCEDURE: All cows were fed an anionic prepartum diet. Blood sample was taken within 6 hours after parturition from randomly selected cows. Thirty-nine cows had a diagnosis of RFM at 24 hours after parturition; 113 were not affected with RFM. At calving, body condition score (BCS; 1 [thin] to 5 [obese]), parity, and calving difficulty score were recorded. Plasma calcium, phosphorus, magnesium, NEFA, BHB, and glucose concentrations were compared between cows with or without RFM. RESULTS: Cows with RFM had significantly lower plasma calcium concentration soon after calving, compared with cows without RFM. Cows with a parity of > or = 3 had significantly lower plasma concentrations of calcium and higher concentrations of magnesium, compared with cows with a parity of 1 or 2. Cows with a BCS of > or = 3.25 at calving had significantly higher plasma concentrations of BHB than cows with a BCS of 2.75 to 3.0. Cows with dystocia had significantly higher plasma concentrations of glucose, compared with cows without dystocia. CONCLUSIONS AND CLINICAL RELEVANCE: In parturient cows fed a prepartum anionic diet, those with RFM have lower plasma calcium concentrations than cows without RFM, although this association does not prove a cause-effect relationship. PMID- 15334841 TI - Development of a questionnaire to measure the effects of chronic pain on health related quality of life in dogs. AB - OBJECTIVE: To develop a reliable, validated questionnaire that can be used for the assessment of chronic pain and its impact on health-related quality of life (HRQL) in dogs. SAMPLE POPULATION: 17 owners of dogs that had chronic pain associated with chronic degenerative joint disease and 165 other dog owners. PROCEDURES: Psychometric methods were used to identify relevant domains, create an item pool, select and validate items, and construct and preliminarily test a structured questionnaire. Relevant domains were identified through semi structured interviews. Descriptor-generating exercises provided the terms owners used to describe these domains and formed an item pool. A selection from this pool was validated and used to construct a questionnaire that underwent preliminary testing. RESULTS: The structured questionnaire contained 109 simple, familiar, descriptive terms associated with good health or chronic pain (most describing subtle aspects of behavior that owners interpreted as expressions of subjective experiences of their dogs) for 13 possible HRQL domains. Each descriptor was associated with a 7-point numeric scale. CONCLUSIONS AND CLINICAL RELEVANCE: The questionnaire was intended to facilitate rapid, sensitive, and accurate rating of a comprehensive range of relevant domains by naive raters with minimal burden on respondents. The principles underlying the development and design of this structured questionnaire offer a novel approach to the proxy measurement of HRQL and changes in HRQL associated with chronic pain for a range of animal species. IMPACT FOR HUMAN MEDICINE: This novel approach may be applicable to other nonverbal populations (eg, young children or elderly people with cognitive impairment). PMID- 15334842 TI - Use of thermal threshold response to evaluate the antinociceptive effects of butorphanol in cats. AB - OBJECTIVE: To characterize the antinociceptive actions of several doses of butorphanol by use of a thermal threshold testing device specifically designed for cats. ANIMALS: 6 domestic shorthair cats. PROCEDURE: The study was a masked, randomized, crossover design. Thermal thresholds were measured by use of a thermal threshold-testing device specifically developed for cats. A small probe containing a heater element and temperature sensor was held with consistent contact against a shaved area of the cat's skin with an elasticized band. Skin temperature was recorded before each test, prior to activation of the heater. On detection of a response (eg, the cat flinched, turned, or jumped), the stimulus was terminated and the threshold temperature recorded. Three baseline measurements were recorded before IV injection of 0.1, 0.2, 0.4, or 0.8 mg of butorphanol/kg. Each cat received all doses in a randomized order at least 1 week apart. The investigator was unaware of the treatment received. Thermal thresholds were measured every 15 minutes for 6 hours. RESULTS: Mean+/-SD pretreatment threshold temperature for all cats was 40.8+/-2.2 degrees C. There were no dose related differences among treatments. There was a significant increase in threshold values for all treatments from 15 to 90 minutes after injection. Mydriasis was detected in all cats after treatment with butorphanol and dysphoric behavior was frequently exhibited. CONCLUSIONS AND CLINICAL RELEVANCE: Results obtained by use of a thermal stimulus indicated that the duration of antinociceptive action of butorphanol was 90 minutes and there was no dose response relationship in cats. PMID- 15334843 TI - Assessment of the influence of fatty acids on indices of insulin sensitivity and myocellular lipid content by use of magnetic resonance spectroscopy in cats. AB - OBJECTIVE: To determine whether dietary fatty acids affect indicators of insulin sensitivity, plasma insulin and lipid concentrations, and lipid accumulation in muscle cells in lean and obese cats. ANIMALS: 28 neutered adult cats. PROCEDURE: IV glucose tolerance tests and magnetic resonance imaging were performed before (lean phase) and after 21 weeks of ad libitum intake of either a diet high in omega-3 polyunsaturated fatty acids (3-PUFAs; n = 14) or high in saturated fatty acids (SFAs; 14). RESULTS: Compared with the lean phase, ad libitum food intake resulted in increased weight, body mass index, girth, and percentage fat in both groups. Baseline plasma glucose or insulin concentrations and glucose area under the curve (AUC) were unaffected by diet. Insulin AUC values for obese and lean cats fed 3-PUFAs did not differ, but values were higher in obese cats fed SFAs, compared with values for lean cats fed SFAs and obese cats fed 3-PUFAs. Nineteen cats that became glucose intolerant when obese had altered insulin secretion and decreased glucose clearance when lean. Plasma cholesterol, triglyceride, and non esterified fatty acid concentrations were unaffected by diet. Ad libitum intake of either diet resulted in an increase in both intra- and extramyocellular lipid. Obese cats fed SFAs had higher glycosylated hemoglobin concentration than obese cats fed 3-PUFAs. CONCLUSION AND CLINICAL RELEVANCE: In obese cats, a diet high in 3-PUFAs appeared to improve long-term glucose control and decrease plasma insulin concentration. Obesity resulted in intra- and extramyocellular lipid accumulations (regardless of diet) that likely modulate insulin sensitivity. PMID- 15334844 TI - Clinical and pathologic comparison of acute leptospirosis in dogs caused by two strains of Leptospira kirschneri serovar grippotyphosa. AB - OBJECTIVE: To develop a method for inducing acute leptospirosis in dogs. ANIMALS: 31 nine-week-old female Beagles. PROCEDURE: Beagles were randomly assigned to 2 inoculation groups or a control group. Dogs were inoculated on 3 successive days by conjunctival instillation of 5 x 10(7) cells of Leptospira kirschneri serovar grippotyphosa strain 82 (12 dogs) or strain RM 52 (14 dogs). Control dogs (n = 5) were similarly inoculated with sterile leptospiral culture media. Clinical signs, clinicopathologic variables, anti-leptospiral antibody titers, and evidence of leptospires in tissues and body fluids were evaluated. Dogs were euthanatized and necropsied on days 7, 14, 22, or 28 after inoculation or as required because of severe illness. RESULTS: Clinical signs in infected dogs included conjunctivitis, lethargy, diarrhea, dehydration, vomiting, and icterus. Consistent clinicopathologic alterations included azotemia, hyperphosphatemia, increased anion gap, hyperbilirubinemia, and an increase in alkaline phosphatase activity. Leptospires were cultured from the kidneys (11/12), urine (6/9), aqueous humor (9/12), blood (12/12), and liver (12/12) of dogs inoculated with strain 82. Only 3 of 14 dogs became infected after inoculation with strain RM 52. Histopathologic lesions in infected dogs included interstitial nephritis, renal tubular degeneration and necrosis, pulmonary hemorrhage, and hepatic edema and perivasculitis. CONCLUSIONS AND CLINICAL RELEVANCE: Conjunctival exposure to L kirschneri serovar grippotyphosa strain 82 resulted in acute leptospirosis in all inoculated dogs, but only 3 of 14 dogs inoculated with strain RM 52 became infected. This method of infection by serovar grippotyphosa can be used to study the pathogenesis and prevention of leptospirosis in dogs. PMID- 15334845 TI - Inhibitory and excitatory neurotransmitters in the cerebrospinal fluid of epileptic dogs. AB - OBJECTIVE: To determine concentrations of excitatory and inhibitory amino acids in CSF of a large number of dogs with idiopathic epilepsy or genetic epilepsy and to evaluate changes in CSF amino acid concentration with regard to drug treatment and sex. ANIMALS: 35 Labrador Retrievers with genetic epilepsy (20 male and 15 female), 94 non-Labrador Retrievers with idiopathic epilepsy (71 male and 23 female), and 20 control dogs (10 male and 10 female). PROCEDURE: Collection of CSF was performed > 72 hours after the occurrence of seizures. Cerebrospinal fluid concentrations of gamma-aminobutyric acid (GABA), glutamate (GLU), aspartate (ASP), serine, and glycine were determined by use of high performance liquid chromatography with electrochemical detection. RESULTS: CSF concentrations of GABA and GLU were significantly lower in Labrador Retrievers with genetic epilepsy (LR-group dogs) than in control-group dogs or in non-Labrador Retrievers with idiopathic epilepsy (non-LR-group dogs). The GLU-to-GABA ratio was significantly higher in LR-group dogs than in non-LR-group dogs. CSF concentrations of GLU and ASP were significantly lower when all dogs with epilepsy (non-LR- and LR-group dogs combined) were compared with control-group dogs. CONCLUSIONS AND CLINICAL RELEVANCE: A decrease in CSF concentrations of GABA appears to play a role in the pathogenesis of genetically determined epilepsy in Labrador Retrievers. However, this decrease in CSF concentrations of GABA may also be a consequence of seizure activity. The GLU-to-GABA ratio may prove to be a useful indicator of genetic epilepsy in Labrador Retrievers. PMID- 15334846 TI - Correlation of helical and incremental high-resolution thin-section computed tomographic and histomorphometric quantitative evaluation of an acute inflammatory response of lungs in dogs. AB - OBJECTIVE: To compare quantitative densitometric computed tomography (CT), morphometric, and histologic data of normal lungs in dogs with similar parameters obtained after induction of an acute inflammatory response and determine whether CT densitometry correlated with histopathologic changes. ANIMALS: 6 healthy adult dogs. PROCEDURE: After initial CT, 1 mL of 0.1 M hydrochloric acid (HCI) and 3 mL of autologous blood were instilled into the right middle (RM) and caudal segment of the left cranial (LCCd) lung lobes, respectively. Immediately and 24 hours after instillation, CT was repeated. At 24 hours, dogs were euthanatized and lungs were fixed and sampled for morphometric and histologic evaluation. The CT data were compared with lung morphology and morphometry by use of unpaired t tests. Comparison with lungs from control dogs was performed using Spearman rank correlation coefficients. RESULT: Mean Hounsfield units (HU) from control and baseline HU from experimental dogs were identical. Immediately after instillation of HCI or blood, there was increased attenuation in both lobes. Autologous blood initially induced severe changes that almost completely resolved at 24 hours; HCI induced severe changes at 24 hours. Significant increases in percentage of parenchymal airspace and alveolar diameter resulted in decreased surface area-to volume ratio in lobes receiving HCl. Histologic scores were significantly higher in the RM lobe, compared with controls. CONCLUSIONS AND CLINICAL RELEVANCE: Computed tomography attenuation correlated well with histomorphometry and histologic findings in this model. Lung lesions after autologous blood were transient and of limited severity. Lesions induced by HCl were severe; alterations in morphometric and histologic parameters were reflected in CT attenuation measurements. PMID- 15334847 TI - Investigation of the effect of acepromazine on intravenous glucose tolerance tests in dogs. AB - OBJECTIVE: To investigate the effects of administration of acepromazine on IV glucose tolerance tests (IVGTTs) in dogs. ANIMALS: 8 male mixed-breed dogs. PROCEDURE: With a 1-week interval between tests, each dog underwent (in random order) an IVGTT with or without pretest administration of acepromazine maleate (0.1 mg/kg, SC, 30 minutes prior to the start of the IVGTT). Food was withheld from the dogs for 14 hours prior to each test. Blood samples were obtained at 20, 10, and 1 minute prior to and at 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 19, 22, 25, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, and 180 minutes after administration of glucose. RESULTS: There were no significant differences in the baseline (ie, after food was withheld) plasma glucose, lactate, and insulin concentrations between dogs undergoing the IVGTT and acepromazine-IVGTT; however, lower baseline free fatty acid concentration was observed in acepromazine-treated dogs. Analysis of data via the application of Bergman's minimal model of glucose kinetics revealed no differences in insulin sensitivity, acute insulin response to glucose, disposition index, or glucose effectiveness between dogs treated or not treated with acepromazine before testing. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that in dogs undergoing IV glucose tolerance testing, pretest administration of small doses of acepromazine can be used as a means of chemical restraint without interfering with results of the glucose metabolism assessment. PMID- 15334848 TI - Relationships between a proprietary index, bispectral index, and hemodynamic variables as a means for evaluating depth of anesthesia in dogs anesthetized with sevoflurane. AB - OBJECTIVE: To evaluate relationships among various techniques for monitoring anesthetic depth in sevoflurane-anesthetized dogs undergoing orthopedic surgery. ANIMALS: 10 dogs. PROCEDURE: Dogs were medicated with acepromazine (0.05 mg/kg, IM), buprenorphine (0.01 mg/kg, IM), and atropine (0.04 mg/kg, IM). Anesthesia was induced and maintained with sevoflurane. Cardiovascular and respiratory responses were monitored. Anesthetic depth was monitored by use of the bispectral index (BIS), and a proprietary index was used to monitor activity of the autonomic nervous system. RESULTS: A significant decrease in BIS was seen after induction but concurrent changes were not observed for the other techniques. The proprietary index increased significantly after intubation, but no changes were seen for the other techniques. No significant changes were detected during incision or when higher nociceptive stimuli were applied. We did not identify a correlation between BIS and the proprietary index, the proprietary index and hemodynamic variables, or the BIS and hemodynamic variables during induction and maintenance. A significant increase in the proprietary index and BIS was detected at the time of resumption of reflexes. During anesthetic recovery, a correlation was found between the proprietary index and BIS but not between hemodynamic variables and the other techniques. CONCLUSIONS AND CLINICAL RELEVANCE: A significant increase in the proprietary index, but not the BIS or hemodynamic variables, was detected during intubation. Anesthetic induction with sevoflurane did not prevent the sympathetic stimulus attributable to tracheal intubation. Monitoring of hemodynamic variables does not provide sufficient information to allow clinicians to evaluate stress during anesthetic recovery. PMID- 15334849 TI - Investigation of the composition, turnover, and thermal properties of ruptured cranial cruciate ligaments of dogs. AB - OBJECTIVE: To assess different components of the extracellular matrix with regard to their thermal properties, composition, and turnover in ruptured cranial cruciate ligaments (CCLs) of dogs, compared with components of intact CCLs from a breed predisposed to CCL failure. SAMPLE POPULATION: Ruptured CCLs obtained from 8 dogs of breeds predisposed to ruptured CCLs and intact CCLs from 12 cadaveric Labrador Retrievers. PROCEDURE: Ruptured and intact CCLs were analyzed for water content; collagen content and collagen cross-links were evaluated via hydroxyproline and amino-acid analyses, respectively. Glycosaminoglycan (GAG) content was analyzed via dimethylmethylene blue and uronic acid assays. Matrix metalloproteinases (MMPs)-2 and -9 and the tissue inhibitors of metalloproteinases (TIMPs)-1 and -2 were detected via gelatin SDS-PAGE zymography and reverse gelatin zymography. Thermal analysis of ligaments was performed by use of differential scanning calorimetry. RESULTS: Ruptured CCLs had significantly higher lamounts of immature cross-links, total and sulfated GAGs, and water content, compared with that of the intact ligaments. Compared with intact CCLs, concentration of pro-MMP-2 was significantly higher in ruptured CCLs; the maximum temperature of collagen denaturation was significantly lower in the ruptured CCLs. CONCLUSIONS AND CLINICAL RELEVANCE: The extracellular matrix of ruptured CCLs had an increased matrix turnover indicated by increased collagen and GAG synthesis, compared with that of intact CCLs. Although the extracellular matrix changes may have occurred before ligament rupture, it is possible that these observed changes may be part of a reparative process after rupture. PMID- 15334850 TI - Distribution of mRNA that codes for subtypes of adrenergic receptors in the gastrointestinal tract of dairy cows. AB - OBJECTIVE: To describe the distribution of mRNA that codes for 9 subtypes of adrenergic receptors in the digestive tract of dairy cows. SAMPLE POPULATION: Fresh full-thickness wall specimens from the abomasum (fundus, corpus, and antrum), ileum, cecum, proximal loop of ascending colon, and 4 locations of the spiral colon collected from 10 healthy cows at slaughter. PROCEDURE: Concentrations of mRNA that code for 9 subtypes of adrenergic receptors in the bovine gastrointestinal tract (alpha1A, alpha1B, alpha1D, alpha2AD, alpha2B, alpha2C beta1, beta2, and beta3) were measured by use of a quantitative real-time reverse transcription-polymerase chain reaction assay. Results were reported in relation to mRNA expression of the housekeeping gene glyceraldehyde phosphate dehydrogenase (GAPDH). RESULTS: Mean mRNA contents of adrenergic receptors in the bovine digestive tract were low (range, 0.00006% to 5.04% of GAPDH). Distribution of receptor subtypes was similar in all tissues, with lowest expression of alpha1D receptors, followed by alpha2B, alpha2C, beta3, alpha1B, alpha1A, beta1, and beta2 in the abomasum, whereas alpha2AD and beta2 in the intestines were highest. In comparison with the intestines, relative concentrations of mRNA for receptors beta2 and beta3 were significantly lower in the abomasum. CONCLUSIONS AND CLINICAL RELEVANCE: Relative concentrations of mRNA that code for adrenergic receptors differed among receptor subtypes and among locations in the bovine gastrointestinal tract. Comparison of these values established in healthy cattle with results for cows with motility disorders, such as abomasal displacement and cecal dilatation, may lead to improved therapeutic or prophylactic approaches for these diseases. PMID- 15334851 TI - Distribution of mRNA that codes for 5-hydroxytryptamine receptor subtypes in the gastrointestinal tract of dairy cows. AB - OBJECTIVE: To describe the distribution of mRNA that codes for 8 subtypes of 5 hydroxytryptamine receptors (5-HTRs) in the digestive tract of dairy cows. SAMPLE POPULATION: Fresh full-thickness wall specimens from the abomasum (fundus, corpus, and antrum), ileum, cecum, proximal loop of ascending colon, and 4 locations of the spiral colon collected from 10 healthy cows at slaughter. PROCEDURE: Concentrations of mRNA that code for 5-HTR subtypes (5-HTR1A. 5-HTR1B, 5-HTR1D, 5-HTR1F, 5-HTR2A, 5-HTR2B, 5-HTR2C, and 5-HTR4) in the bovine digestive tract were measured by use of a quantitative real-time reverse transcription polymerase chain reaction assay. Results were reported in relation to mRNA expression of the housekeeping gene glyceraldehyde phosphate dehydrogenase (GAPDH). RESULTS: Mean relative mRNA concentrations for 5-HTR were low (range, 0% to 1.32% of GAPDH), and mRNA that codes for 5-HTR1A was not detected. In the abomasum, mRNA expression was highest for 5-HTR1B and 5-HTR2B, followed by subtypes 1F 2A, 1D, and 4, whereas 5-HTR2C was not detected. In intestinal samples, concentrations of subtypes 1B, 2B, and 4 were highest, followed by 1D, 1F, 2A, and 2C. Relative concentrations of mRNA that code for 5-HTR2A were significantly higher in the abomasum than the intestines, but lower for 5-HTR2B, 5-HTR2C, and 5-HTR4. CONCLUSIONS AND CLINICAL RELEVANCE: Relative concentrations of mRNA that code for 5-HTRs differ among locations in the gastrointestinal tract of cattle. Understanding differences in the distribution of 5-HTRs in healthy cattle and cattle with gastrointestinal tract disease may lead to improved therapeutic approaches for abomasal and cecal motility disorders. PMID- 15334852 TI - Evaluation of three-dimensional ultrasonography of the bovine mammary gland. AB - OBJECTIVE: To evaluate 3-dimensional (3-D) ultrasonography of the bovine mammary gland, particularly of the teat. ANIMALS: 6 lactating cows. PROCEDURES: Clinical and ultrasonographic examinations of mammary glands of each cow were performed. Teats were removed from a slaughtered cow and examined via ultrasonography. All scans were performed by use of a sensorless standard transducer (8.5 to 10 MHz linear array). The 2-dimensional data acquired were downloaded to an off-line system, and software was used to digitize each image and produce a 3-D block of digitized information. The selected anatomic area was displayed as a 3-D volume cube. RESULTS: Good-quality, 3-D views of the entire mammary gland were acquired by obtaining sections through the glandular parenchyma, gland cistern, teat cistern, and teat canal. CONCLUSIONS AND CLINICAL RELEVANCE: 3-D ultrasonography allows perspective images of the mammary gland to be obtained. Many scanners routinely used in veterinary practice have the resolution required to produce satisfactory images, and the cost of refitting a standard ultrasonographic unit with 3-D software is affordable; however, the cost of a volume transducer with a positioning system and mechanical sweep strategy may be prohibitive. Three dimensional ultrasonography is a new imaging technique that has promising applications in many fields of veterinary medicine. PMID- 15334853 TI - Comparison of L-selectin and Mac-1 expression on blood and milk neutrophils during experimental Escherichia coli-induced mastitis in cows. AB - OBJECTIVE: To evaluate L-selectin (CD62L) and Mac-1 (CD11b) expression at the surface of blood and milk neutrophils during the early inflammatory response to Escherichia coli-induced mastitis in cows. ANIMALS: 6 healthy Holstein heifers in early lactation. PROCEDURE: Blood and milk samples were collected before and after intramammary administration of 10(4) CFU's of E coli in the left mammary gland quarters. Bacterial counts and electrolyte concentrations in milk, rectal temperature, differential blood leukocyte counts, milk somatic cell counts, neutrophil viability, and the expression of CD62L and CD11b on blood and milk neutrophils were determined longitudinally. RESULTS: Bacteria grew during the first 6 hours after inoculation with a pronounced leukocytic influx. Coincident with neutrophil influx was an increase in CD62L+ and CD11b+ milk neutrophils, as well as an improved viability of milk neutrophils. The peak of the inflammatory reaction was reached approximately 12 hours after E coli inoculation. From that time forward, changes in CD62L and CD11b expression were opposed to each other, with a decrease in CD62L expression and an increase in CD11b expression on blood and milk neutrophils; the magnitude of the differences in CD62L and CD11b expression between blood and milk neutrophils decreased. Percentages of CD62L+ and CD11b+ milk neutrophils increased to percentages that were similar to blood neutrophils (ie, approx 92%). CONCLUSIONS AND CLINICAL RELEVANCE: The presence of adhesion molecules on a large percentage of milk neutrophils during the acute inflammatory response, together with the changes in receptor density, suggest a major role for CD62L and CD11b in neutrophil function during coliform mastitis. PMID- 15334854 TI - The inevitability of change. PMID- 15334855 TI - Heparin overview and issues. AB - Unfractionated heparin (UFH), which has been available commercially for over half a century, has been the most widely used agent for quickly suppressing thrombosis. When given intravenously, UFH quickly binds to and activates antithrombin, which then inhibits several activated factors in the clotting cascade. For decades, UFH was invaluable for treating arterial and venous thrombosis, and no alternative was available. The short half-life of UFH and the fact that its action could be reversed readily with protamine made it an almost ideal antithrombotic agent. However, variable pharmacokinetics, together with problems of inaccuracy and unreliability of the activated partial thromboplastin time, have made it difficult to use this drug optimally. In addition, side effects such as osteoporosis, heparin-induced thrombocytopenia (HIT), and delayed HIT have led to increased concerns about the use of UFH in view of the advantages offered by newer agents. Fractionating heparin into low-molecular-weight heparins that still retain the pentasaccharide active site provided a way to achieve the same type of therapeutic effect with more predictable dosing and fewer adverse effects. Similarly, a pentasaccharide has been synthesized and marketed as fondaparinux. Although these advances have improved our therapeutic options, continued advances on the horizon raise the question of whether the use of UFH will soon be abandoned. PMID- 15334856 TI - Challenges in variation and responsiveness of unfractionated heparin. AB - Unfractionated heparin (UFH) has been in clinical use for more than half a century. Despite its undoubted contribution to the treatment and prevention of thrombosis, heparin is significantly limited by its variable biochemical composition and unpredictable pharmacokinetics. The situation is compounded by the fact that methods for monitoring heparin do not necessarily reflect its therapeutic effect. The activated partial thromboplastin time (aPTT) is a method for monitoring heparin therapy that is simple, cheap, and readily available. However, it is also poorly standardized and is affected by numerous factors-both analytic and preanalytic-that are unrelated to the heparin effect. Establishing an appropriate therapeutic range for the aPTT is challenging for smaller clinical laboratories, and the antifactor Xa method of measuring heparin levels is not widely available. The College of American Pathologists published consensus guidelines in an effort to improve the laboratory monitoring of UFH therapy. However, it seems unlikely that the laboratory problems associated with monitoring UFH will be resolved. Unfractionated heparin is highly antigenic and carries a significant risk of heparin-induced thrombocytopenia (HIT). Even in the absence of thrombocytopenia or thrombosis, the presence of heparin-associated antibodies may predict adverse clinical outcomes and strengthen the rationale for the ultimate replacement of UFH. Fortunately, alternatives to UFH, such as low molecular-weight heparins, direct thrombin inhibitors, and more specific factor Xa inhibitors, are becoming available for clinical use. The pharmacokinetics of these agents are more predictable and rely much less on laboratory monitoring. Nonheparin agents also eliminate the risk of HIT. The emergence of these newer anticoagulants makes the continued use of UFH increasingly difficult to justify. PMID- 15334857 TI - Anticoagulant prophylaxis in medical patients: an objective assessment. AB - Venous thromboembolism (VTE) is a clinically silent and potentially fatal disease that manifests as deep vein thrombosis (DVT) and pulmonary embolism. Venous thromboembolism remains a serious public health challenge, with an ever increasing odds ratio of occurrence given the aging population in the United States. This article reviews the epidemiology of VTE; risk factor identification and stratification as a means of advancing awareness, prevention, and detection of VTE; and prophylaxis options and their outcomes, particularly administration of unfractionated heparin (UFH) 5000 U subcutaneously every 12 versus 8 hours in the at-risk medical patient population. The important studies comparing outcomes of these different UFH dosing regimens compared with placebo and low-molecular weight heparins also are discussed. Consensus recommendations shaping contemporary clinical practice guidelines in this setting are highlighted. A systemwide approach to treatment of all medical patients who are risk stratified and receiving appropriate pharmacologic prophylaxis is recommended. PMID- 15334858 TI - Treatment of venous thromboembolism: challenging the unfractionated heparin standard. AB - Venous thromboembolism (VTE) is a major public health problem leading to high morbidity and mortality in the United States. Since more then 50% of patients with VTE may have asymptomatic disease, the start of appropriate therapy often is delayed. Traditionally, intravenous unfractionated heparin (UFH) has been used to manage the acute phase of VTE. Although an effective agent, numerous limitations are associated with the use of UFH therapy, such as the need for careful monitoring and frequent dosing adjustments. In addition, the assay used to monitor UFH--the activated partial thromboplastin time (aPTT)--does not correlate reliably with plasma heparin levels or antithrombotic activity. In the early 1990s, the low-molecular-weight heparins (LMWHs) emerged as alternative anticoagulants to UFH and began to successfully challenge the UFH standard for treatment of VTE. Clinical evidence has consistently demonstrated that LMWHs given subcutaneously are at least as safe and as effective, if not better, than intravenous UFH. This anticoagulant class has a much more predictable dose response relationship, requires little or no monitoring, and provides cost-saving opportunities for outpatient management of VTE. The LMWHs are now considered the treatment of choice for many patients with VTE and are largely replacing UFH for this indication. In the last decade, additional agents, such as direct thrombin inhibitors and factor Xa inhibitors, have emerged as potential future alternatives for treatment of VTE. As clinical data regarding these new agents for treatment of VTE continue to evolve, their role in clinical practice will be elucidated. PMID- 15334859 TI - Unfractionated heparin in cardiology: redefining the standard of practice. AB - Acute coronary syndromes (ACS) consist of unstable angina (UA), non-ST-segment myocardial infarction (NSTEMI) and ST-segment myocardial infarction (STEMI). Timely intervention with effective, predictable antithrombin therapy is critically important in the early management of these conditions. Platelet aggregation is also an important component of thrombus formation in arterial thrombosis. Historically, unfractionated heparin (UFH) has been combined with aspirin to suppress thrombin propagation and fibrin formation; however, its effectiveness has been questioned in this setting. Unlike newer anticoagulant alternatives, UFH paradoxically stimulates platelet aggregation, which may further promote clot formation. In addition, obtaining a valid therapeutic activated partial thromboplastin time (aPTT) in cardiology patients is a major challenge, and dosing is complex. Due to substantial variation in reagents and instruments, target aPTT ranges for UFH in ACS clinical trials cannot be extrapolated to individual institutions. Further, the risk of ischemic events is greater shortly after abrupt discontinuation of UFH compared with alternative agents with longer half-lives and less stimulation of platelet aggregation. Key UA-NSTEMI clinical trials have demonstrated that UFH is inferior to newer agents, such as the low-molecular-weight heparins (LMWHs). Consistent with this evidence, the most recent practice guidelines of the American College of Cardiology and the American Heart Association in UA-NSTEMI identify the LMWH enoxaparin as the agent of choice. In patients with STEMI receiving the fibrinolytic tenecteplase as reperfusion therapy, enoxaparin has also been superior to UFH in combination. In percutaneous procedures, newer indirect (enoxaparin) and direct (bivalirudin) antithrombins have demonstrated safety and efficacy. There is little doubt that as we move forward in optimizing adjunctive anticoagulation in the cardiology setting, UFH will largely be replaced by better antithrombin agents. PMID- 15334860 TI - Unfractionated heparin dosing nomograms: road maps to where? AB - Weight-adjusted nomograms have been a significant advance in the use of unfractionated heparin (UFH). Clinical trials have demonstrated the ability of weight-adjusted nomograms to achieve a therapeutic activated partial thromboplastin time (aPTT) more rapidly than with standard UFH dosing. Despite this advantage, a significant number of patients have subtherapeutic and supratherapeutic aPTTs. Real-world experiences also corroborate the inability to keep UFH therapeutic with the use of nomograms. Despite the limitations of UFH nomograms, they have been used in several different types of venous and arterial thrombosis treatment settings. Unfortunately, these nomograms are not all consistent and require a considerable amount of time for training health care professionals on their use in order to limit the potential for medication errors. Although UFH nomograms have provided advancement over standard UFH dosing, their limitations still generate the desire for a more predictable anticoagulant. PMID- 15334861 TI - Unfractionated heparin: focus on a high-alert drug. AB - Unfractionated heparin (UFH) is associated with a high rate of drug-related problems due to either its inherent pharmacologic properties or an extension of these properties often caused by medication errors. The drug-related problems associated with UFH can significantly hinder the success of therapy and negatively affect the overall cost of care. Unfractionated heparin has been classified as a high-alert drug by the Institute for Safe Medication Practices. Approximately 2.1% of the total records submitted to the MedMARx national error database were related to UFH; 4.5-5.5% of these errors reported were harmful. With this high potential for error, it is essential that all health care providers adopt a collaborative or systems approach to identify solutions to reduce the occurrence of these medication errors. The Joint Commission on Accreditation of Healthcare Organizations has published national patient safety goals for improving the safety of patient care, many of which are applicable to UFH therapy. Unfractionated heparin drug-related problems not necessarily related to medication errors include heparin-induced thrombocytopenia, bleeding events, and osteopenia. Heparin-induced thrombocytopenia is a serious complication of heparin therapy and remains seriously undiagnosed. Bleeding events often occur with therapeutic as well as prophylactic UFH administration even when monitoring indexes are within the therapeutic range. However, due to the variability associated with UFH monitoring methods, definitive guidelines are lacking to assist in avoiding such serious events. Osteopenia has been associated with long term UFH therapy; one third of patients experience reductions in bone density, potentially leading to fractures. Today, safer alternative anticoagulation therapies are available, such as the low-molecular-weight heparins. When compared with UFH, these alternative therapies provide equivalent or superior efficacy for numerous indications. PMID- 15334862 TI - Unfractionated heparin: a nursing dilemma. AB - Nurses face challenges in all aspects of their practice, especially with administering and monitoring drugs in a safe, effective manner. Key factors known to affect drug administration include general drug knowledge, formal nurse education, continuing education needs, clinical experience, and the nationwide nursing shortage. Other factors are advances in technologic aids and quality improvement initiatives. Emphasis on patient safety is growing, especially as it relates to drug therapy and high-alert drugs such as unfractionated heparin (UFH). Specific interventions related to UFH administration can enhance patient care management. Because nurses are at the site of direct patient care, they are often in an opportune position for identifying medication errors. At the same time, and most important, nurses need to collaborate with other health care professionals to actively develop solutions to minimize these errors. Adopting a systems approach and working collaboratively with an interdisciplinary team can result in improved patient outcomes. PMID- 15334863 TI - Economics of unfractionated heparin: beyond acquisition cost. AB - Despite numerous clinical trials demonstrating advantages of new anticoagulants, unfractionated heparin (UFH) is still used by a number of clinicians in the United States. The reason for this continued use of UFH is not superior efficacy, improved safety, or convenience, but low acquisition cost. However, several other costs associated with the use of UFH are often not considered. Appropriate economic analysis, which considers both cost and outcomes, has not demonstrated support for continued use of UFH. Its continued use based simply on lower cost is not justified by the literature. PMID- 15334864 TI - Prevention of paclitaxel-associated arthralgias and myalgias. PMID- 15334865 TI - Management of thrombosis in the cancer patient. AB - The treatment of venous thromboembolism in the cancer patient presents many challenges. The safety and efficacy of anticoagulation in these patients are still evolving. The clinical scenario is often unique to each patient and dictates careful consideration of the method of anticoagulation. Various issues that need to be considered are the length of anticoagulation, the method of administration, and the potential pitfalls of initiating one therapy over another. This review attempts to provide an approach to these issues in the treatment of the cancer patient with venous thromboembolism. The role of warfarin, heparin, and low-molecular-weight heparin is discussed. Additionally, adjunctive measures, such as vena cava filter insertion and thrombotic complications associated with central venous catheters, are discussed. The review is meant to provide clinicians with practical guidelines through a review of the pertinent literature. PMID- 15334866 TI - Promising new approaches to managing mucositis. PMID- 15334867 TI - Erythropoietic agents in the management of cancer patients. Part 1: Anemia, quality of life, and possible effects on survival. AB - Erythropoietic agents have been shown to be well tolerated and highly effective in correcting the anemia associated with cancer. Studies aimed at optimizing their use in this regard are ongoing and are evaluating the potential value of early dose intensification through "front-loading" schedules, the feasibility of reducing the frequency of dosing, and the value of early initiation of erythropoietic agents to prevent anemia. The potential for erythropoietic therapy to contribute to survival in cancer patients also is being investigated. Anemia is an independent poor prognostic factor in several cancer settings, such as head and-neck, lung, and cervical cancer. In patients with B-cell chronic lymphocytic leukemia, it is possible that the period of stable disease--that is, postponement of the need for cytotoxic chemotherapy--may be prolonged when patients are "downstaged" through correction of their anemia. In addition, anemia may affect the patient's quality of life in ways that impact compliance and/or the ability to tolerate therapy. Retrospective analysis of large phase III trials designed to evaluate the effect of erythropoietic agents on anemia in cancer patients has reported a trend toward improved survival among patients who received erythropoietic therapy. Trials to evaluate how correction of anemia in cancer patients might impact survival are ongoing. In this first installment of a two part article, the author reviews the clinical data supporting the use of erythropoietic agents for the management of the fatigue due to anemia, the effect of anemia control on quality of life, and the evidence available so far that control of such anemia may actually lengthen survival. In the next issue, Dr. Smith will discuss the possible role of erythropoietic agents in neuroprotection and neurotherapy, including cognitive dysfunction in cancer patients. PMID- 15334868 TI - Incidence and management of hot flashes in prostate cancer. AB - Hot flashes are as common in men who have been castrated due to prostate cancer as hot flashes are in women after menopause. The symptom can cause significant discomfort for a considerable length of time. The hot flashes are most likely caused by a reduction in sex-hormone levels, which, in turn, causes an instability in the hypothalamic thermoregulatory center. Calcitonin gene-related peptide is involved in menopausal hot flashes in women and possibly also in castrated men. The mainstays of treatment for castrated men with hot flashes remain estrogens, progesterone, and cyproterone acetate, each of which has different side effects. Other treatments for hot flashes include clonidine and antidepressants and, according to one uncontrolled study, electrostimulated acupuncture. Nonetheless, there is a need for more effective and less toxic treatments. In this review, we will discuss the prevalence, duration, distress, physiology, and treatment options of hot flashes in men subjected to castration therapy due to prostate cancer. PMID- 15334869 TI - Glutamine does not prevent paclitaxel-associated myalgias and arthralgias. AB - Myalgias and arthralgias are prominent toxicities associated with paclitaxel. Pursuant to pilot information suggesting that glutamine could markedly alleviate this toxicity, we developed a placebo-controlled, double-blind, randomized crossover trial to test this hypothesis. This trial studied 36 patients who had experienced myalgias/arthralgias related to a prior paclitaxel-containing regimen and who were then randomized to receive oral glutamine (10 grams three times a day, starting on the day of chemotherapy) x 5 days or an identical-appearing placebo. For the subsequent cycle of chemotherapy, patients were crossed over, again in a double-blind manner, to receive the alternative treatment. Patients recorded daily myalgia/arthralgia scores on numerical scales. Physicians recorded patient-reported myalgias/arthralgias using National Cancer Institute common toxicity criteria at baseline and after each course of therapy. The results of this study revealed that glutamine had no effect on the development or severity of paclitaxel-induced myalgias/arthralgias, as recorded either by the patients' daily logs or by physician-reported information. Glutamine was well tolerated, with no suggestion of more toxicity compared to placebo. Upon completion of the two cycles, patients were asked to indicate which of the two blinded courses they preferred. Of those patients indicating a preference, 29% preferred the glutamine cycle, compared with 33% who preferred the placebo cycle (P = 0.96). Thus this trial did not suggest any role for oral glutamine, compared with placebo, for preventing the development, or alleviating the severity, of paclitaxel-induced myalgias/arthralgias. PMID- 15334870 TI - Is there evidence for earlier intervention in cancer-associated weight loss? PMID- 15334871 TI - Ketamine in the treatment of refractory cancer pain: case report, rationale, and methodology. PMID- 15334872 TI - Pathogenesis of cancer cachexia. AB - Cachexia is a progressive wasting syndrome characterized by extensive loss of adipose tissue and skeletal muscle. It occurs in about half of all cancer patients. While anorexia also may be present, the energy deficit alone does not explain the pathogenesis of cachexia. The presence of an acute phase response (APR) has been linked to accelerated weight loss and a shortened survival time. The APR is thought to be initiated by cytokines such as interleukin (IL)-6 and IL 8, production of which is induced by a tumor factor, proteolysis inducing factor (PIF). Cachectic cancer patients also show an increased expression of uncoupling protein-3 in muscle, which may act as an energy sink, increasing energy expenditure. Loss of adipose tissue appears to be due to an increase in degradation of triglycerides, rather than a decrease in synthesis. One candidate for this effect is a tumor lipid mobilizing factor, which stimulates lipolysis directly through a cyclic AMP-mediated process via interaction with a beta3 adrenergic receptor. Loss of skeletal muscle arises from both a depression in protein synthesis and an increase in protein degradation. The major proteolytic pathway involved in intracellular protein breakdown in cachectic muscle is the ATP-ubiquitin-dependent proteolytic pathway. Both PIF and tumor necrosis factor alpha, but not other cytokines, can induce expression of the key regulatory components of this pathway. Eicosapentaenoic acid, found in oily fish, effectively attenuates protein degradation in cachectic muscle by inhibiting the increased proteasome expression and can stabilize body weight in cachectic cancer patients. PMID- 15334873 TI - Communication skills: myths, realities, and new developments. PMID- 15334874 TI - Management of cancer pain in geriatric patients. AB - The management of cancer in the older person has become a central issue of oncology. At present, some 60% of all cancers in the United States and approximately 70% of cancer-related deaths occur in persons aged 65 and older, and these figures will continue to increase at a significant rate as the number of older people in our population grows. This review studies the management of pain, one of the most common and consequential symptoms of cancer and cancertreatment, in the older person. In particular, the article explores the influence of age on pain perception, assessment, and management. These questions will be addressed after attempting a definition of age in biological terms. In the conclusion, the American Geriatrics Society's guidelines for the management of pain in older individuals will be examined and a research agenda outlined. PMID- 15334876 TI - Communication and the patient/physician relationship: a phenomenological inquiry. AB - All interaction between patient and medical team is mediated through communication. In working with advanced cancer patients, troubled communication between the patient and healthcare team is sometimes evident. The current study examined the perspective of seven advanced cancer patients on this communication. The patients were under the care of an urban cancer research center, and each had expressed, at least once, a desire for hastened death. Using a phenomenological form of inquiry, serial, "in-depth semistructured" interviews were conducted (mean = 3; range = 2-6) at a place convenient to the patient. The interviews were audiotaped, transcribed, coded, and organized into themes. The study findings suggest that the most important aspects of communication between the advanced cancer patient and medical team at an urban cancer research center are the extreme vulnerability of the patient to both style and content of the communication; the extreme sensitivity of the communication itself, because it concerns issues vital to the patient; and the complexity of the communication, encompassing and expressing both verbal and non-verbal messages, as well as intended and unintended messages. Conflict inherent in almost all communication between patient and medical team involved disagreement about goals of care and the participants' perception of not being treated with respect. Harm caused to the participant by communication missteps included loss of hope, a sense of abandonment, and diminished feelings of personal dignity and worth. The findings may not be representative of advanced cancer patients being cared for in a different treatment setting. PMID- 15334875 TI - Management of thalidomide toxicity. AB - Thalidomide has re-emerged as a novel antineoplastic agent with immunomodulatory and antiangiogenic activities. In the early sixties, it was withdrawn from the market after its infamous association with congenital abnormalities that left about 10,000 children affected world-wide. With strict regulations and precautions, thalidomide is now approved by the FDA for the treatment of erythema nodosum leprosum. Its role in cancer therapy is promising, with clinical trials in the past 5 years showing significant activity in multiple myeloma. Several trials are ongoing in other malignancies, such as myelodysplastic syndrome, agnogenic myeloid metaplasia, renal cell carcinoma, and prostate cancer. The major toxicities of thalidomide are birth defects, sensorimotor peripheral neuropathy, somnolence, rash, fatigue, and constipation. Less common side effects include deep venous thrombosis, Stevens-Johnson syndrome, elevated liver enzymes, malaise, and peripheral edema. The incidence and severity of adverse events are related to dose and duration of therapy. Doses of the drug of 200 mg/day or less are usually well tolerated. In this review, we will discuss the incidence and management of the side effects of thalidomide and the precautions and interventions needed to minimize the toxicities of this drug. PMID- 15334877 TI - Preventing radiation-induced cystitis with hyaluric acid. PMID- 15334879 TI - Influence of paroxysmal atrial fibrillation attack on brain natriuretic peptide secretion. AB - OBJECTIVES: Plasma brain natriuretic peptide (BNP) concentration is higher during atrial fibrillation (Af) than sinus rhythm, based on studies of electrical defibrillation treatment of patients with chronic Af. However, the change in paroxysmal atrial fibrillation (PAf) is not well known. This study investigated such changes and the relationship between BNP and Af. METHODS: BNP levels were successfully measured at three time points: before Af attack, during Af attack, and after (spontaneous or pharmacological) termination of 68 consecutive Af attacks in 35 outpatients with PAf (23 men, 12 women, mean age 70.4 +/- 9.6 years). BNP was measured by immunoradiometric assay. RESULTS: BNP (median[quartiles]) during PAf was increased by 66[25, 120] pg/ml (2.4-fold) compared to during sinus rhythm (p < 0.0001), and fell to the former level after return to sinus rhythm (before attack = 39[18, 70], during attack = 102[52, 205], after attack = 35[20, 67]). BNP increased in 55 (81%) of 68 attacks, did not change (within +/- 20 pg/ml) in 11 (16%), and decreased in 2 (3%). BNP was already elevated immediately (within 4 hr) after onset of Af, and BNP elevation (delta BNP) showed no significant relationship with the time elapsed after onset. During the Af attack, 41% of PAf patients were asymptomatic although BNP increased significantly. CONCLUSIONS: These results suggest that elevated amounts of BNP during Af are released from secretory granules in the atrium, and BNP elevation of unknown cause may be attributed to the presence of asymptomatic Af. Cardiac function evaluation using BNP during Af requires special consideration, unlike during sinus rhythm, even in patients with PAf or chronic Af, because BNP during Af is the sum of the BNP values from the ventricle (reflecting left ventricular function) and the atrium (due to Af). PMID- 15334878 TI - Prescribing methadone, a unique analgesic. PMID- 15334880 TI - Effect of short-term administration of high dose L-arginine on restenosis after percutaneous transluminal coronary angioplasty. AB - BACKGROUND: A single and local administration of L-arginine after balloon angioplasty enhances nitric oxide (NO) generation and inhibits lesion formation in animals. OBJECTIVES: The present study assessed the effect of increasing NO to inhibit restenosis after percutaneous transluminal coronary angioplasty (PTCA) in humans by local and systemic administration of L-arginine, a precursor of NO in humans. METHODS: L-arginine was administered to 34 consecutive patients with angina pectoris or old myocardial infarction via a cardiac catheter (500 mg/4 min) before PTCA, and via a peripheral vein (30 g/4 hr, for 5 days) after PTCA. Patients were treated between December 1998 and December 2000. Plasma concentrations of L-arginine, NO (as nitrite + nitrate) and cyclic guanosine monophosphate (cGMP) were measured before and after L-arginine administration. The control group consisted of 90 patients who underwent PTCA successfully without L-arginine administration in the period between July 1996 and November 1998. Baseline clinical and angiographic characteristics were compared between the two groups. All patients were followed by coronary angiography for 3 months after PTCA. Quantitative coronary angiography and restenosis rate were studied. RESULTS: Baseline clinical and angiographic characteristics were not different between the two study groups. Despite a significant elevation in plasma L arginine concentration after L-arginine administration, NO and cGMP did not increase significantly. After PTCA, the difference in restenosis rates between L arginine and control subjects (34% vs 44%) was not significantly different. CONCLUSIONS: Short-term administration of high dose L-arginine did not significantly change the restenosis rate after PTCA. PMID- 15334881 TI - [Isolated left ventricular noncompaction in the elderly: a case report]. AB - A 83-year-old man with an implanted pacemaker was admitted to our hospital for dyspnea. Echocardiography revealed left ventricular dilation and diffuse hypokinesis mimicking dilated cardiomyopathy. Multiple trabeculations and deep intertrabecular recesses were prominent in the apical and posterior walls. Computed tomography with contrast medium also showed prominent trabeculations. Thallium-201 and iodine-123-beta-methyl-p-iodophenyl-pentadecanoic acid scintigraphy demonstrated fixed defects in the inferior wall. These findings were consistent with isolated noncompaction of the ventricular myocardium. The patient's symptoms of heart failure improved in response to treatment, including carvedilol. Isolated noncompaction of the ventricular myocardium is recognized as a rare congenital disorder of infancy with fatal outcomes, with some reports of cases in adults. The present patient is the most elderly described to date. PMID- 15334882 TI - [Uremic pericarditis complicating cardiac tamponade: a case report]. AB - A 29-year-old man developed diabetes mellitus in 1983 and diabetic nephropathy which gradually worsened from 1998. He was admitted to our hospital for initiation of peritoneal dialysis in May 2002. However, the efficiency of dialysis was not sufficient to improve elevated levels of blood urea nitrogen and serum creatinine. His body weight and cardiothoracic index by chest roentgenography gradually increased starting 9 days after admission. To improve the efficiency of dialysis, we tried to increase the dialysis fluid. Nevertheless, the efficiency of peritoneal dialysis remained low, and the patient complained of nausea 14 days after admission. Hypotension suddenly occurred 16 days after admission. Echocardiography showed massive pericardial effusion and collapse of the right ventricle. The diagnosis was cardiac tamponade. We performed cardiac centesis and pericardial drainage which revealed bloody pericardial effusion. Urgent hemodialysis was performed. The differential diagnosis of cardiac tamponade was established. After hemodialysis, the amount of pericardial effusion decreased, the gastro-intestinal symptoms disappeared, and the blood urea nitrogen and serum creatinine levels decreased. We speculated that the cause of cardiac tamponade was uremic pericarditis after ruling out infectious disease, collagen disease, malignant disease, and aortic dissection. Cardiac tamponade due to uremic pericarditis has become very rare since hemodialysis was developed. PMID- 15334883 TI - [Vegetative cardiac metastases of oral cavity cancer: an autopsy case report]. AB - A 60-year-old man was admitted to our hospital for evaluation of intracardiac vegetative masses detected by echocardiography in September 2001. He had undergone surgery for oral cavity cancer in 1999. He presented with severe embolic symptoms including cerebral infarction, but had few symptoms of heart failure. Antibiotic therapy was started under the diagnosis of infective endocarditis, but the embolic symptoms persisted. An autopsy revealed that the intracardiac vegetative masses consisted of tumor cells originating from the oral cavity cancer. Intravascular tumor thrombi were also found widely distributed in other organs such as the liver, lung, spleen and kidney, and had similar histological features. This is a very rare case of cardiac metastases of oral cavity cancer without adhesion to the endocardium or other myocardial tissue. PMID- 15334884 TI - [A 55-year-old man presenting with manifest ST elevation 16 hours after reperfusion therapy for acute anterior myocardial infarction]. PMID- 15334885 TI - [A 57-year-old man with intracardiac tumor occurring after operation for rupture of Valsalva sinus]. PMID- 15334886 TI - [Guidelines for exercise eligibility at schools, work-sites, and sports in patients with heart diseases (JCS 2003)]. PMID- 15334887 TI - Role of the urinary calcium in the growth of calcium stone. AB - We analyzed the relationship between the rate and clinical factors. The growth rate per year of the stone was measured by Nabeshima's method in 29 male patients with renal calcium stones including 7 pure calcium oxalate (CaOx) stones and 22 mixed calcium oxalate and calcium phosphate (CaOx-CaP) stones. The 24-hour urinary excretion of calcium, phosphate, uric acid and magnesium were assayed under an ambulatory free diet in 5 patients with CaOx stones and 15 with CaOx-CaP stones. The relationship between the growth rate and the urinary excretion of stone-forming parameters was examined. We found a significant positive correlation between the growth rate of calcium stones and the urinary excretion of calcium (p<0.02). In addition, the growth rate of CaOx-CaP stone was significantly higher than that of pure CaOx stone (p<0.05). In conclusion, urinary calcium is important for the growth of renal calcium stones. PMID- 15334888 TI - Clinical course in patients with percutaneous nephrostomy for hydronephrosis associated with advanced cancer. AB - We investigated the clinical courses of 33 patients with advanced malignancies who has undergone percutaneous nephrostomy (PNS) to clarify the efficacy and validity of PNS and evaluated the results according to the type of catheter. The pigtail, Malecot and balloon catheters were used. All the catheters were exchanged every 4 weeks, if necessary for larger ones. The site of the primary disease, preoperative performance status, duration of time spent at home after PNS construction, number of times the catheter was changed, type and size of the catheter and complications associated with PNS were compiled. The median overall survival was 3.0 months. Twenty-three patients (69.7%) were never discharged from hospital. Thirty patients died of their primary diseases. The median overall survival of patients with cancers of the upper gastrointestinal system, large bowel, urological organs and gynecologic organs were 1.5, 5.5, 3.0 and 3.0 months, respectively (p=0.0116). Eighteen patients (54.5%) experienced complications such as pyelonephritis, obstruction, dislodgement and so on. PNS is an appropriate urinary diversion for patients with hydronephrosis with advanced cancer and unilateral construction is effective for treating end-stage obstructive renal failure. However, PNS rarely improves the patient's quality of life and the risk of complications is unavoidable regardless of the type of catheter used. PMID- 15334889 TI - [The effect of radiotherapy on patients with prostate specific antigen failure after radical prostatectomy]. AB - We studied the effect of radiotherapy on patients with re-elevation of prostate specific antigen (PSA) after radical prostatectomy. Radiotherapy was performed on 8 patients with re-elevated PSA after radical prostatectomy without any previous treatment. Pathological stages were B in 2 patients, and C in 6 patients. Patients received three-dimensional dynamic conformal radiotherapy, and irradiation doses ranged from 44 to 70 Gy (median 60). The target area of irradiation included prostatic bed and seminal vesicles. PSA levels before radiotherapy were 0.31-1.9 ng/ml (median 0.40). In 7 patients, PSA levels decreased and no increase has been observed thereafter. In one patient, PSA level increased during radiotherapy; therefore, the treatment was discontinued at 44 Gy. Two patients suffered grade 1 to 2 acute toxicities, and no late toxicity has been observed so far. Radiotherapy is considered one of the effective treatments for re-elevation of PSA after radical prostatectomy. PMID- 15334890 TI - [Intravesical Bacillus Calmette-Guerin (BCG: Tokyo 172 strain) instillation for carcinoma in situ of the bladder: results with 6 successive instillations of 40 mg BCG]. AB - We performed a study to evaluate the usefulness of intravesical Bacillus Calmette Guerin (BCG: Tokyo 172 strain) instillation on carcinoma in situ (CIS) of the bladder. Between 1998 and 2003, 43 patients were treated for CIS of the bladder with a median follow-up period of 45 months (range: 12 to 69 months). The patients (35 males and 8 females) ranged in age from 45 to 89 years (average: 67.5 years). They underwent intravesical instillation of 40 mg of BCG once a week for 6 weeks. A complete response (CR) was achieved in 83.7% of the patients. Among these patients, 97.2% and 70.7% remained recurrence-free during follow up for one year and three years, respectively. The median duration of CR was 31.5 months. Although total cystectomy was performed on 1 patient, none of the patients died of bladder cancer. Adverse effects included bladder irritability in 48.8%, pyuria in 46.5%, gross hematuria in 18.6%, and fever (temperature over 37.5 degrees C) in 9.3%. No clinically significant side effects were observed. These results indicate that intravesical instillation of BCG at a dose of 40 mg given 6 times was as effective as the routine dose of 80 mg, and could decrease systemic adverse effects such as high fever. PMID- 15334891 TI - [Metastatic adrenal carcinoma suspected as a primary tumor: a case report]. AB - A 62-year-old man was admitted with chief complaints of fever and general fatigue. Enhanced computed tomography revealed a tumor (5 cm in diameter) in the right suprarenal space, and the right renoportal lymph nodes were swollen. An abdominal TI-weighted MR image showed a low-intensity tumor measuring 4.5 x 5.5 x 6.0 cm. Chest computed tomography revealed two tumors. One was 1.5 cm, on the hilum of left lung, the other was 1 cm in the S6 lung field near the pleura. These findings strongly indicated primary adrenal carcinoma and lung metastasis. Right adrenalectomy was performed. Histological examination of this tumor revealed diffuse tumor cells with irregular nuclei, and it was diagnosed as poorly differentiated adenocarcinoma. There was no possibility of primary adrenal carcinoma. The primary site of the tumor was suspected to be the left lung. PMID- 15334892 TI - [Postoperative hypoglycemia after resection of pheochromocytoma: a case report]. AB - We report a case of severe hypoglycemia following resection of pheochromocytoma. A 39-year-old male was admitted to our hospital with a chief complaint of palpitation. Blood pressure and fasting blood glucose were within the normal range. Computed tomography and magnetic resonance imaging revealed a right adrenal tumor (7.5 x 5 x 7 cm) and 131I-MIBG scintigraphy showed marked tumor uptake of isotope. As plasma and urinary catecholamine levels were very high, a clinical diagnosis of pheochromocytoma was established. Doxazosin and propranolol were administered for 43 days prior to adrenalectomy. During intensive care monitoring 2 hours postoperatively, the patient became drowsy and began to sweat. Although blood pressure remained stable, severe hypoglycemia (38 mg/dl) and hyperinsulinism (63.67 microU/ml) were confirmed. Infusion of 50% glucose improved these symptoms and plasma glucose level and insulin secretion normalized within 15 days of surgery. We also reviewed 25 cases of hypoglycemia after resection of pheochromocytoma. We recommend close monitoring of blood glucose for at least 6 hours after adrenalectomy for pheochromocytoma. PMID- 15334893 TI - Giant hydronephrosis with increased carbohydrate antigen 19-9 both in serum and fluid. AB - We report a case of right giant hydronephrosis. A 68-year-old man was admitted to our hospital with chief complaints of general fatigue, loss of appetite and a one year history of progressive fullness on whole abdomen. Abdominal computed tomography scan exhibited a huge, homogeneous, low density mass originating from the right kidney. We performed right percutaneous nephrostomy and drained over 6,500 ml bloody fluid. Cytological examination of the drained fluid revealed atypical nuclear appearance defined as class III. Increased values of carbohydrate antigen 19-9 were observed both in the fluid as well as in the serum. We performed right nephrectomy. Macroscopic appearance of the resected kidney showed marked stenosis at the portion of ureteropelvic junction. Histological analysis of the stenotic portion demonstrated marked fibrosis without findings of malignancy. PMID- 15334894 TI - [A case of abdominal desmoid tumor that was difficult to differentiate from urachal tumor]. AB - A 76-year-old male patient complained of frequent urination. Computed tomographic and magnetic resonance imaging scans revealed a heterogeneous mass involving his bladder, prostate and Sigmoid colon. He was diagnosed with an invasive urachal tumor and surgery was performed due to severe tumor adhesion to adjacent organs. Partial resection of the tumor was performed with partial cystectomy and sigmoidectomy. A post-operative pathological diagnosis revealed the tumor was a desmoid abdominal wall tumor. The desmoid abdominal wall tumor was not easily differentiated preoperatively from an urachal tumor. PMID- 15334895 TI - [A case of urachal xanthogranuloma suspected to be a urachal tumor]. AB - A 47-year-old female consulted our hospital with the chief complaints of lower abdominal pain and fever. There was a palpable mass in the lower abdomen. The patient had undergone oophorectomy by lower abdominal median incision. Ultrasonography, computed tomography, and magnetic resonance imaging (MRI) demonstrated a cystic mass above the bladder dome extending to the umbilicus, which was strongly suspected to be a urachal tumor. Enhanced T1 weighted MRI showed a mass enhanced by contrast media. Partial cystectomy with urachal resection was performed by lower abdominal median incision. The histological diagnosis was xanthogranuloma. This is the thirteenth case report of xanthogranuloma of the urachus in the Japanese literature. PMID- 15334896 TI - [A case of retrovesical leiomyoma]. AB - We herein report a rare case of leiomyoma in the retroperitoneal space posterior to the urinary bladder. A 61-year-old man came to our department complaining of lower abdominal discomfort. Abdominal and pelvic computed tomographic scan revealed a retrovesical solid tumor on the cranial side to the left seminal vesicle. Diagnostic imaging suggested that the retrovesical tumor was a benign tumor such as leiomyoma or fibroma, and he underwent simple resection of this retrovesical tumor via reroperitoneal approach. Histopathological diagnosis was well compatible with image diagnosis of leiomyoma. He has been followed up for 6 months without recurrence. PMID- 15334897 TI - [A case report of idiopathic hypogonadotropic hypogonadism with particular reference to monitoring of prostate volume]. AB - A 23-year-old man visited our hospital, complaining of a lack of secondary sexual characteristics. Based on both clinical and endocrinological examinations, he was diagnosed as suffering from idiopathic hypogonadotropic hypogonadism (IHH). With human chorionic gonadotropin (hCG)/human menopausal gonadotropin (hMG) replacement therapy, clinical symptoms and serum testosterone levels improved to normal ranges. Interestingly, prostate volume as measured by transrectal ultrasonography increased considerably from 3.6 ml before treatment to 20.2 ml after treatment. The monitoring of prostate volume might be a parameter useful for evaluating the treatment effect of hCG/hMG replacement therapy in IHH. PMID- 15334898 TI - [A case report: testicular pure seminoma metastasized to costal bone after 2 years post-operatively]. AB - A 32-year-old man underwent orchiectomy for his right testicular tumor (pure seminoma, pT1, stage A(I)). Pelvic and para-aortic lymph nodes were irradiated with 18 Gy as adjuvant therapy. Two years later, he developed low back pain. Computed tomography, magnetic resonance imaging and bone scans showed an enhanced mass at the 10th left costa. Aspiration cytology showed seminoma. After administration of chemotherapy with bleomycin, etoposide and cisplatinum, following high dose chemotherapy with peripheral blood stem cell transplantation, the costal lesion was diminished and symptoms relieved. Then radical costectomy was performed. A histopathological study showed non-viable cells of seminoma. Post-operative progress was uneventful. Testicular pure seminoma with bone metastasis is rare, and to our knowledge, only 9 cases have been reported. PMID- 15334899 TI - Multicystic malignant mesothelioma of the tunica vaginalis with an unusually indolent clinical course. AB - We report an extremely rare case of a multicystic malignant mesothelioma in the tunica vaginalis with an unusually indolent clinical course. A 48-year-old man presented with a one-month history of painless swelling of right scrotal contents. Ultrasonography and computed tomography (CT) revealed a multicystic mass in the right scrotal sac with evidence of neither distant nor lymph node metastases. The testicular tumor markers were within the normal limits. Inguinal orchiectomy was performed under the suspicion of a malignant tumor. The cystic tumor consisted of fibrocellular, microcystic and adenomatoid elements microscopically was diagnosed biphasic malignant mesothelioma of tunica vaginalis but no invasion into the testis, epididymis and also scrotum. The patient has been disease-free for 72 months and is being followed on an outpatient basis with no further adjuvant therapy. PMID- 15334901 TI - Chromatography in the conservation of cultural heritage. PMID- 15334900 TI - Erosion of an inflatable penile prosthesis due to autoinflation. AB - A 59-year-old man presented with erosion of an inflatable penile prosthesis. The cylinders were not deflated and the right one protruded from the urethra. Autoinflation that causes failure of deflation is an annoying and embarrassing mechanical failure. This failure may cause intraurethral erosion of the prosthesis. Urologists should inform their patients who hope for implantation of an inflatable prosthesis of this potential adverse event prior to surgery. PMID- 15334902 TI - Characterisation of proteinaceous binders in artistic paintings by chromatographic techniques. AB - This review discusses the application of chromatographic techniques (GC, HPLC and Py-GC) for the characterisation of proteinaceous materials in artistic paintings. The focus is on the various analytical steps that are needed to determine these natural materials in paint samples, from sampling and sample pre-treatment, including various methods of hydrolysis and derivatisation for GC and HPLC, to approaches for data evaluation. PMID- 15334903 TI - Characterisation and identification of proteinaceous binding media (animal glues) from their amino acid profile by capillary zone electrophoresis. AB - A capillary electrophoretic method for identifying different species of proteinaceous binders--collagen, egg white, and milk casein--is described. It allows characterisation of the proteins on the basis of the amino acid profiles obtained after their acidic hydrolysis. The profiles of the underivatised amino acids are recorded directly by capillary zone electrophoresis at pH 2.26 with the aid of a conductivity detector, thus eliminating the need for a derivatisation step. Identification is carried out by means of different relative peak areas of the amino acids. A scheme is given for identification, which is based on the main markers, hydroxyproline, proline, glycine, glutamic acid, and serine and valine. PMID- 15334904 TI - Impact of biological factors on binding media identification in art objects: identification of animal glue in the presence of Aspergillus niger. AB - The materials and especially organic materials used for creation of art objects can be utilized by various microorganisms for their growth and facilitate the microbial colonization of the object. An understanding of the chemical alterations in artefacts caused by the presence of microorganisms can be crucial for correct identification of the materials initially used for the artefact creation--nowadays an important step in restoration and/or art-historical investigation of the art object. The present article describes a model experiment in which we investigated the possible chemical alterations in animal glue films used as substrate for growth of the fungus Aspergillus niger. The sterilized animal glue solution was poured into Petri dishes, inoculated with Aspergillus niger, and subsequently incubated at 15 degrees C for 0, 7, 9, 14, and 28 days. After interruption of incubation, the content of the Petri dish was analyzed for amino acid composition by the GC-MS based method. It was found that the growth of Aspergillus niger on animal glue films did not cause significant changes in the amino acid composition of the film and had no impact on animal glue identification. PMID- 15334905 TI - Microbial diversity in biodeteriorated monuments as studied by denaturing gradient gel electrophoresis. AB - Studies of the microorganisms living on cultural assets and their potential damaging effects begin with an understanding of the microbial diversity present in such samples. Recently, molecular techniques have started to attract considerable interest since they do not require the culture of microorganisms in order to detect their presence. The basic protocol followed during microbial diversity analysis on cultural heritage assets utilizes electrophoretic techniques for separating highly homologous sequences. At present, Denaturing Gradient Gel Electrophoresis (DGGE) is the most frequently reported technique for separating DNA fragments during microbial diversity studies of art objects. In this report, we describe DGGE profiling techniques, show the most common applications, and present some examples of their use in microbial diversity studies on cultural heritage. PMID- 15334906 TI - Effects of traditional processing methods of linseed oil on the composition of its triacylglycerols. AB - Different oil processing methods were performed, which included washing with water and treatment with lead-based driers, with and without heating to different temperatures, giving a set of 7 oils to be investigated. The effects of the traditional processing methods of linseed oil on its triacylglycerol (TAG) composition were studied, using the following analytical methods: high performance size exclusion chromatography (HPSEC), Fourier transform infrared spectroscopy (FTIR), high-performance liquid chromatography-atmospheric pressure chemical ionisation-mass spectrometry (HPLC-APCI-MS), direct temperature resolved mass spectrometry (DTMS), matrix assisted laser desorption/ionisation time-of flight mass spectrometry (MALDI-TOF-MS), and electrospray ionisation Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS). A decrease of the initial cis-double bonds and the formation of trans-double bonds upon heating of the oils was observed. Heating a lead and oil mixture to 150 degrees C, or heating the oil alone to 300 degrees C led to the highest degree of oxidation. A difference was observed for the oxidation patterns for oils with and without the addition of lead. Furthermore, levels of oxygen incorporation were higher when lead was added to the oil. High temperature treatment of the oils resulted in an increased average molecular weight. The changes in the initial conformation of the double bond systems observed with FTIR were supported by HPLC-APCI-MS measurements that showed the formation of a number of new isomeric TAGs in the heated oil compared to freshly pressed, untreated oil. Oligomerisation up to hexamers was observed with HPSEC, and MALDI-TOF-MS. The formation of oligomers up to trimers only, however, was observed with ESI-FTICR-MS. Incorporation of oxygen was mainly observed with MALDI-TOF-MS and ESI-FTICR-MS whereas with DTMS and FTIR hardly any evidence was found for this. PMID- 15334907 TI - Direct chemolysis-gas chromatography-mass spectrometry for analysis of paint materials. AB - A novel direct method using (m-trifluoromethylphenyl)trimethylammonium hydroxide (TFTMAH) has been developed for on-line (trans)esterification of lipidic materials (drying oils, egg) prior to GC-MS analysis. The method was first optimised by comparing three N-methylammonium hydroxide reagents (TMAH, PhTMAH, TFTMAH) and triolein as a lipid standard. Secondly, the procedure was tested on a series of fresh and naturally aged (up to 40 years) (un)pigmented drying oils and egg yolk reference film standards. Thirdly, the method was applied to the analysis of a sample from an altar painting "Resurrection of Christ" by Francesco Solimena (1723) from the Chapel of Upper Belvedere in Vienna. The relative proportions of the fatty acids from the translucent size layer confirm the presence of partially degraded oil, probably prepolymerised walnut oil. PMID- 15334908 TI - Identification of "insoluble" red dyewoods by high performance liquid chromatography-photodiode array detection (HPLC-PDA) fingerprinting. AB - The paper describes a high performance liquid chromatography-UV/Vis spectrometry detection analytical approach to the identification of some redwood species of historical importance in textile dyeing. The group of extracted dyestuffs considered as "insoluble" because of their non-aqueous or alkaline extraction conditions is present in the wood of the Pterocarpus family and Baphia nitida species. First, the crude extracts of tinctorial and related species and their chromatographic fingerprints were studied. This part of work shows that some species not yet mentioned in the literature have potential dyeing properties. Subsequent experiments performed on the redwood cargo of a 200-year-old archaeological shipwreck allowed identification of the water-logged wood species. Furthermore, the different methods of dyestuff extraction used for dyeing according to traditional recipes and their impact on analytical results were studied. They show that standard recovery obtained by acid hydrolysis of dyestuff from dyed yarns is inadequate. Hence, alternative solvent-based procedures were proposed. The identification of species in textile threads then becomes possible. The applied approach was validated by analysis of dyed reference yarns with some indications of crude material extraction mode. The employed method of analysis seems to be useful for "insoluble" wood species identification in cultural heritage artifacts as well as for phytochemical purposes, despite the fact that very few detected color compounds were chemically identified. PMID- 15334909 TI - Characterisation of embalming materials of a mummy of the Ptolemaic era. Comparison with balms from mummies of different eras. AB - Gas chromatography-mass spectrometry has been used to determine the nature of organic materials used in mummification balms. A comparative analysis of samples taken from Egyptian mummies is developed. The results are given in two parts. First, it is shown that the chemical composition of the balm is practically independent of the part of the mummy from which it is taken. This study was done on a Ptolemaic mummy (circa 100 BC from the Guimet Museum in Lyon). Fats, beeswax, and diterpenic resins were the main components: they were found everywhere. Castor oil was also very often detected (in half of the samples). This particular fat is present in the balm inside the thorax but not in the skull. Moreover it is shown that a vegetable tannin was employed. Components indicative of vegetable tannin input (gallic acid and inositols) were found in seven samples out of eighteen, particularly close to the body and on the canopic pack of the heart. Secondly, some conclusions from a comparative study of the composition of balms from mummies of various social levels as well as of different Egyptian periods are reported. It is shown that beeswax was used as from very early times (XVIIIth dynasty). The mixture of beeswax, fats, and diterpenoid resins would appear to be more recent. The balms of three mummies dating from more recent Egyptian periods (XIXth to XXVth dynasty) were analysed. No evidence of a resin, gum-resin, or plant gum could be found. Some mummies would appear to have been embalmed with fats or beeswax. Finally, the entrails canopic pack said to belong to Ramses II undoubtedly shows an embalming process with a triterpenic resin of the mastic type. The adopted analytical methodology enabled us to achieve simultaneous detection of four components of the balm of the Ptolemaic mummy. Analysis of the other five mummies revealed far less complex chemical compositions for the balms. This may be an indication of different embalming processes, although we should bear in mind the question of organic matter preservation through the ages. PMID- 15334910 TI - Headspace solid phase microextraction for screening for the presence of resins in Egyptian archaeological samples. AB - The aim of this study was to use headspace solid phase microextraction (SPME) to reveal the presence of resin in archaeological samples, such as mummification balms, from ancient Egypt. Experiments were first performed with fresh resins of known origin. The SPME fibre readily extracted mono- and sesquiterpenes and, to a lesser extent, diterpenes. Using mass spectra and retention indices of constitutive compounds, qualitative analysis of the volatile fraction allowed us to differentiate resins or gum-resins such as myrrh, olibanum, galbanum, labdanum, mastic, and conifer resins. SPME was then successfully applied to archaeological samples from ancient Egypt in which the presence of resins was detected. Volatile components were desorbed and trapped according to the same SPME procedure as was applied to fresh resins, after a sample preparation consisting of a fine grinding. PMID- 15334911 TI - Investigating the history of prehistoric glues by gas chromatography-mass spectrometry. AB - Although organic materials are very sensitive to biochemical alteration processes, they may be preserved for millennia in various archaeological contexts. Remains of adhesives made during prehistory were discovered at different sites, in the form of residues adhering to flint tools and ceramic vessels or as free lumps in sediment. To characterise the natural substances exploited for adhesive production during late prehistory, we undertook GC and GC/MS analysis of 90 samples from 8 sites dating from the Neolithic to Iron Age periods. This paper discusses our approach to the study of organic adhesives preserved in archaeological contexts, with a particular focus on the presentation of the various categories of organic adhesives that we analysed and the choice of chromatographic conditions adapted to the specificity of such samples. The results obtained show that birch bark tar, a triterpenoid adhesive made by destructive distillation of white birch bark, was predominantly used during the neolithic period even though other materials such as various barks or organic fossil substance were also used. During the Bronze and Iron ages, which follow the Neolithic period, adhesive production is evolving through the expansion of the range of the natural substances used (identification of diterpenoid pine resin) and the addition of beeswax as a plasticiser to birch bark tar. By combining chromatographic analysis and archaeological data, it was thus possible to follow the evolution of adhesive making at the end of prehistory, testifying to the inventiveness of the craftsmen whatever the period considered. PMID- 15334912 TI - Whatever happened to capillary electrochromatography? PMID- 15334913 TI - Separation techniques for the analysis of artists' acrylic emulsion paints. AB - Emulsion paints are complex chemical systems. The main problems in their characterization are related to the similarities in polymer composition and to the presence of many different types of additives. Thus high resolution separations, sensitivity, and response specificity are required to identify simultaneously the polymer matrix and the minor components. Especially surfactants and pigments are thought to affect significantly the properties of the paint layers during ageing and their identification is the first step in evaluating the behaviour of these products in working conditions. Representative samples of acrylic emulsion paints for artists have been investigated by pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) and size exclusion chromatography-Fourier transform infrared spectroscopy (SEC-FTIR), and the results have been compared. All the analysed samples were bound in ethyl acrylate methyl methacrylate, poly(EA-co-MMA), or n-butyl acrylate-methyl methacrylate, poly(nBA-co-MMA), copolymers. Two types of nonionic surfactants, ethoxylated fatty alcohols and alkylaryl polyethoxylates, commonly used as emulsifiers and dispersive agents have been identified, together with a number of organic pigments. PMID- 15334914 TI - Human genomic DNA analysis using a semi-automated sample preparation, amplification, and electrophoresis separation platform. AB - The growing importance of analyzing the human genome to detect hereditary and infectious diseases associated with specific DNA sequences has motivated us to develop automated devices to integrate sample preparation, real-time PCR, and microchannel electrophoresis (MCE). In this report, we present results from an optimized compact system capable of processing a raw sample of blood, extracting the DNA, and performing a multiplexed PCR reaction. Finally, an innovative electrophoretic separation was performed on the post-PCR products using a unique MCE system. The sample preparation system extracted and lysed white blood cells (WBC) from whole blood, producing DNA of sufficient quantity and quality for a polymerase chain reaction (PCR). Separation of multiple amplicons was achieved in a microfabricated channel 30 microm x 100 microm in cross section and 85 mm in length filled with a replaceable methyl cellulose matrix operated under denaturing conditions at 50 degrees C. By incorporating fluorescent-labeled primers in the PCR, the amplicons were identified by a two-color (multiplexed) fluorescence detection system. Two base-pair resolution of single-stranded DNA (PCR products) was achieved. We believe that this integrated system provides a unique solution for DNA analysis. PMID- 15334915 TI - Analysis of basic compounds at high pH values by reversed-phase liquid chromatography. AB - Reversed phase high performance liquid chromatography (RPLC) is currently the method of choice for the analysis of basic compounds. However, with traditional silica materials, secondary interactions between the analyte and residual silanols produce peak tailing which can negatively affect resolution, sensitivity, and reproducibility. In order to reduce these secondary interactions, which comprise ion exchange, hydrogen bonding, and London forces interactions, chromatographic analyses can be carried out at low or high pH values where silanol groups and basic compounds are mostly uncharged. The chromatographic behaviour of a particular bidentate stationary phase, Zorbax Extend C18, was studied with a set of basic and neutral compounds. Thanks to a higher chemical stability than traditional silica based supports, analyses were carried out with a high pH mobile phase, which represents a good alternative to the acidic mobile phases generally used to reduce ion exchange interactions. The performance of this bidentate stationary phase was also compared with that of other supports and it was proved that it is advantageous to work with high pH mobile phases when analyzing basic compounds. PMID- 15334916 TI - Rapid and high-resolution analysis of geometric polyprenol homologues by connected octadecylsilylated monolithic silica columns in high-performance liquid chromatography. AB - A Chromolith Performance octadecylsilyl (ODS) monolithic silica column (Merck) was compared with a conventional microparticulate ODS-bonded silica column in the high-performance liquid chromatography separation of natural polyprenols. A system comprising two connected monolithic columns afforded an equivalent separation at half the analysis time of the conventional method. Furthermore, ten connected columns achieved a tremendously high-resolution separation, in which the complicated series of homologous polyprenols with geometric isomerism were fully separated. PMID- 15334917 TI - Temperature optimization for improved determination of phosphatidylserine species by micro liquid chromatography with electrospray tandem mass spectrometric detection. AB - A sensitive method for determination of disaturated phosphatidylserine species in the presence of their monounsaturated analogs has been developed, using micro liquid chromatography coupled to electrospray ionization tandem mass spectrometry. The hydrophobic nature of the phosphatidylserine species required a combination of low-eluting sample solvents and sub-ambient temperatures in order to focus large sample volumes up to 20 microL. The samples were dissolved in 2 propanol:hexane:water (20:10:4, v/v/v) prior to 1:9 dilution with ammonium formate buffer:2-propanol:tetrahydrofuran (30:55:15, v/v/v) and final 1:4 dilution with ammonium formate buffer (10 mM):2-propanol: tetrahydrofuran (55:37.5:7.5, v/v/v). The analytical column was a 0.5 x 150 mm stainless steel column packed with 5 microm C30 particles, while the mobile phase contained ammonium formate buffer (10 mM): 2-propanol:tetrahydrofuran (30:55:15, v/v/v). A temperature program from 5 degrees C (hold for 3 minutes) to 75 degrees C at 8 K/min provided separation of the disaturated phosphatidylserine species from their monounsaturated analogs, making available a sensitive determination of the isobaric species. The mass limit of detection for dipalmitoyl phosphatidylserine was 100 pg, corresponding to a concentration limit of detection of 5 pg/microL when using an injection volume of 20 microL. This is an improvement by a factor of 20 as compared to previously reported numbers obtained with conventional LC columns. The within-assay precision of dipalmitoyl phosphatidylserine was 11.9% RSD (n = 3), while the retention time precision was 4.1% RSD (n = 6). PMID- 15334918 TI - Use of binary and ternary mobile phases in reversed-phase high performance liquid chromatographic separation of chiral compounds. AB - The influence of the nature of typical organic modifiers, used in reversed-phase systems diluted with water, on retention and selectivity of chiral acetylated diamines and acetonitriles possessing pharmacological activity was investigated. Linear, semi-logarithmic relationships between the logarithm of the retention factor, k, of the enantiomer and the volume fraction, phi, of the organic modifier in the binary aqueous-organic eluent were established for chiral stationary phases. The slope of a plot of log k vs. the modifier volume fraction depends not only on the chemical structure of the chromatographed enantiomers, but also on the nature of the modifier. Moreover, the enantioselectivity of the studied racemates also depends on the nature and concentration of the modifier and is higher for a methanol mobile phase than for acetonitrile. PMID- 15334919 TI - Hydrocarbon type analysis by thin-layer chromatography with flame-ionization detection: vacuum gas oils, heavy feeds, and hydroprocessed products. AB - Thin-layer chromatography with flame-ionization detection (TLC-FID) provides quantitative hydrocarbon type data as well as distribution of aromatics by ring number. This method has been applied to obtain amounts of saturates, aromatics, and polars in heavy oil distillates such as light vacuum gas oils and heavy vacuum gas oils derived from different crude sources. TLC-FID chromatograms and resultant quantitative hydrocarbon type data show that these distillates vary markedly in aromatic contents and aromatic ring types. Similar observations are made with several fluid catalytic cracking feeds. Effects of process parameters such as operating pressure and temperature on hydroconversion of aromatics and polars from a heavy oil are assessed by TLC-FID. It has been demonstrated that there is a preferential reduction of higher polycyclic aromatic hydrocarbons and polars with an increase of both hydrogen partial pressure and reactor temperature. PMID- 15334920 TI - Recognition of hepatotoxic homologues of Microcystin using a combination of selective adsorption media. AB - Microcystin, a hepatotoxin produced by cyanobacteria, was chromatographically discriminated with newly prepared selective adsorption media. Since the name Microcystin refers to up to 70 different homologues, we have developed a novel technique to prepare selective recognition sites, based on the "Fragment Imprinting Effect". The homologues of Microcystin can be individually discriminated from each other by antigen-antibody assay as well as by C18-based HPLC. This is mainly due to differences in the three dimensional structure of Microcystin homologues; however, differences in toxicity are relatively small among the homologues. Therefore we have to measure the total amount of Microcystin homologues present to determine contamination by Microcystins. We have demonstrated the first group recognition for Microcystin homologues through combination of two different selective adsorption media prepared by a fragment imprinting technique. PMID- 15334921 TI - Determination of volatile fatty acids in landfill leachates by ion-exclusion chromatography. AB - An ion-exclusion chromatographic method with on-line desalinization for the determination of volatile fatty acids in landfill leachates is described. Highly sensitive conductivity detection of the organic acids was achieved by using dilute p-hydroxybenzoic acid solution as an eluent. Interference with mineral acids was reduced by treatment with barium chloride solution prior to desalinization. A silver-loaded cation-exchange guard column for the desalinization was installed in series with the analytical column to avoid the contamination of organic acids. This method features detection limits of 0.01 mg L(-1) formic acid, 0.02 mg L(-1) acetic acid, 0.05 mg L(-1) propionic acid, and 0.1 mg L(-1) butyric acid, respectively, with an injection of 20 microL sample. Application of the on-line desalinization LC method is illustrated for leachate samples from a Japanese sanitary landfill. PMID- 15334922 TI - A modified HPLC method for the determination of aspartic acid racemization in collagen from human dentin and its comparison with GC. AB - The D/L ratio of aspartic acid enantiomers in proteins of low turnover is generally accepted as a reliable procedure for age determination. In our study, twelve samples of eyetooth dentin were analyzed for age determination. The pure insoluble collagen isolated from eyetooth dentin was obtained by an EDTA demineralization process. Free amino acids obtained after collagen hydrolysis were converted into o-phthaldialdehyde-N-acetyl-L-cysteine (OPA-NAC) derivatives for HPLC analysis under modified conditions and into trifluoroacetic acid isopropyl esters for GC analysis, respectively. The modified HPLC procedure used phosphate buffer and acidified sample matrix prior to injection which resulted in suppression of peak tailing of both diastereomers, thus allowing achievement of both good selectivity and good resolution. To ensure the high accuracy of the developed method the other parameters, i.e. specificity, precision, linearity, LOD, and LOQ, were also determined. Nine collagen samples covering the age range of 18 to 84 years were used for the determination of coefficient of racemization (KR) and calculation of parameters for age estimation. The regression equations for the data set analyzed were as follows: KR= 0.0005 x age + 0.0262 (R2 = 0.9639) for HPLC, and KR= 0.0006 x age + 0.0319 (R2 = 0.9374) for GC, respectively. PMID- 15334923 TI - Approaches for the coating of capillary columns with highly phenylated stationary phases for high-temperature GC. AB - Two highly phenylated tetramethyl-p-silphenylene-diphenylsiloxane copolymers were coated on fused silica capillary columns and used as stationary phases in GC. The copolymers offered new insights into the coating process and column preparation due to their physicochemical properties. The fused silica capillary surface had to be pretreated in various ways to achieve a homogeneous film and a well deactivated surface: etching with ammonium bifluoride; leaching with sodium hydroxide and hydrochloric acid; silylation with tetraphenyldimethyldisilazane and triphenylsilylamine. Droplet formation was observed on tetraphenyldimethyldisilazane silylated surfaces leading to capillary columns with low separation efficiency. The topology of inhomogeneous films was investigated by light microscopy, scanning electron microscopy, and Auger electron spectroscopy. It became apparent that the stationary phase did not form droplets but islands, which are connected by a wetting layer according to the Stranski-Krastanov growth mode. Both copolymers are potential stationary phases for high-temperature GC with promising properties. They offer a higher overall polarity than 75% phenyl, 25% methyl-polysiloxanes in combination with increased thermal stability and reduced bleed levels. PMID- 15334924 TI - [500 cases of external ultrasound-assisted liposuction]. AB - OBJECTIVE: To discuss the therapeutic effects and safety of external ultrasound assisted liposuction and to find out the rules of circumference variation in different body areas after liposuction. METHODS: External ultrasound liposuction was performed on 500 patients (595 sites). We recorded the volume of anesthetic drugs and the aspirated pure fat as well as the body contour variations of every area at one to three months postoperatively. The decreased dimension at different body sites after liposuction were compared and analyzed. RESULTS: The volumes of the anesthetic liquid and the aspirated pure fat were the most in the waist abdomen and the thigh. After liposuction, the circumstances in different body areas were all decreased to certain degrees. Along with the severity of obesity, changes in the waist-abdomen became more obvious and the decreases of the umbilical circumference, the minimum and the maximum waist circumferences were prone to be less. But when the, minimum waist circumference was over 111 cm, obvious changes occurred in the umbilical. Meanwhile, the upper-middle part of the thigh and the upper arm showed larger variations than the lower part. The ultrasound-assisted liposuction had less blood loss and pain. It resulted in smooth skin without severe complications. CONCLUSIONS: External ultrasound assisted liposuction is a safe, effective and easily acceptable operation for body contour remodeling. The body circumference variation may have its intrinsic rule, which, to some extent, is meaningful for conducting clinical inquiry and forecasting the operation results. PMID- 15334925 TI - [The swine surgical expermental study of laser-assisted liposuction]. AB - OBJECTIVE: To evaluate the advantage and disadvantage of laser-assisted liposuction compared with conventional liposuction. METHODS: Ten swines were devided into three groups, the laser liposuction group, conventional liposuction group and control group. We compared the two surgical groups with the following aspects: ecchymosi, edema, lipocrit study, hemoglobin studies and blood biochemical changes, etc. RESULTS: There is a benefit of laser-assisted liposuction in the following aspects such as ecchymosi, edema, lipocrit and postoperative complications. The two aspects, that is, ecchymosi and lipocrit study was statistically significant. Hemoglobin change was not statistically significant between the two groups. Laser-assisted liposuction did not lead to dysfunction of organs such as liver and kidney. CONCLUSIONS: Laser-assisted liposuction can significantly decrease the blood lose, ecchymosis and edema compared with conventional liposuction, and it did not give rise to dysfunction of organs. PMID- 15334926 TI - [Effects of the site and method of liposuction on differentiation of human preadipocytes]. AB - OBJECTIVE: To illustrate the effects of the site and method of liposuction on differentiation of human preadipocytes. METHODS: Forty-two fatty samples were obtained with liposuction, which were then divided into four groups according to operation sites (abdomen, hip or extremity) and the methods (conventional negative-pressure or syringe method). Each sample was treated with collagenase I to release preadipocytes for in vitro culture. Affected by the differentiation induced agents for 9 days or 28 days, the cultured adipocytes were stained with Nile red and observed under a fluorescent microscope. The differentiation rates were examined with flow cytometric analysis and the quantity of intracytoplasmic lipids was determined with oil red O staining. The results were analyzed with independent samples t-test (Mann-Whitney) using SPSS 10.0. RESULTS: There was no statistical difference in differentiation at the 9th day or 28th day among the preadipocytes obtained from the abdomen, hip or extremity with the negative pressure method. The preadipocytes from the abdomen differentiated more at the 28th day than the 9th day (P < 0.05), which was not observed in the hip or the extremity. The preadipocytes obtained from the abdomen with the negative-pressure method differentiated more than those with the syringe method (P < 0.05). CONCLUSION: No essential difference was found in preadipocyte differentiation among the liposuction sites, while the abdomen might have some superiority. The negative-pressure method of liposuction is the first option in future research of tissue engineering. The flow cytometric analysis is a convenient way to study preadipocyte differentiation. PMID- 15334927 TI - [Individual prefabricated titanium implant for the reconstruction of the skull bone defect]. AB - OBJECTIVE: This paper presents a new method of individual prefabricated titanium implant for the reconstruction of the skull bone defect. METHOD: A computer-based 3D model of the cranial bone defect is created from helical CT-data and serves as the basis for the computer aided design and manufacturing (CAD/CAM) of the individual prefabricated titanium implant for the cranial bone defect reconstruction. RESULT: Since 2001, a total of nine patients suffering from the cranial bone defect have been operated on by this method with satisfied result. The fallow-up is 3 to 12 months. CONCLUSION: These individual prefabricated titanium implants have won a high precision, a good biomechanics and a excellent biocompatibility. It is a quite ideal and very simple method with much less complication for the surgical treatment of the cranial bone defect. PMID- 15334928 TI - [Malar reconstruction in the patients with Treacher-Collin's syndrome]. AB - OBJECTIVE: The key feature of Treacher-Collin's syndrome is malar dysostosis. The article focused on malar reconstruction for Treacher-Collin's syndrome and compared the implant materials. METHODS: From 1994 to 2002, a total of 55 patients with Treacher-Collin's syndrome were treated with malar reconstruction. In the operation, the lateral orbital rim and the mala were exposed by the bicoronal incision or the subciliary incision. The mala was augmented and reconstructed with implants of different materials, including autologous bone (rib, ilia or cranium). Medpor biomaterial or bone cement. RESULTS: The operations of the 55 patients were all successful without infection. The satisfactory rate in facial contour was 90%. Implant exclusion occurred in 2 cases using hone cement. CONCLUSION: Malar reconstruction is the most important treatment for Treacher-Collin's syndrome. Every implant material has advantages and shortcomings. Autologous hone is the best material for malar reconstruction. Medpor is the best artificial material, with good histocompatibility, without exclusion, absorption and donor injury. PMID- 15334929 TI - [Chin augmentation with bone transplantation from the mandible]. AB - OBJECTIVE: To explore a new method for mentoplasty. METHODS: The bilateral prominent mandibular angle or outer lamina was resected through the intraoral approach. The resected bone fragments were shaped and rigid fixed to the chin with miniplates and screws. RESULTS: A total of 30 patients (28 females, 2 males) accepted chin augmentation with this method. The mandibular angle bone was used in 20 cases and the mandibular outer lamina was used in 10 cases. The operative results were satisfactory, and the patient's facial contour was improved substantially. CONCLUSION: No rejection reaction was found after this procedure. Chin augmentation with autogenous mandibular bone is an ideal method for genioplasty. PMID- 15334930 TI - [Transplantation of free latissimus dorsi myocutaneous flaps for craniomaxillofacial reconstruction]. AB - OBJECTIVE: This clinical study was to improve the surgical treatment to craniomaxillofacial tissue defects. METHODS: Since 1997, eight cases with severe craniomaxillofacial defects were treated using free latissimus dorsi myocutaneous flaps. In the operation, nerve anastomosis was performed. Of the 8 cases, 7 were treated in one stage, 1 was treated in 3 steps. The craniomaxillofacial defects ranged from 10 cm x 8 cm to 30 cm x 12 cm. The flaps was 12 cm x 10 cm to 32 cm x 16 cm in size. RESULTS: Postoperative follow-up for 6 months to 4 years demonstrated satisfactory results in all the cases. There was neither necrosis nor ulcer after the operation. The sensation recovery of the flap was also satisfactory. CONCLUSION: Free transfer of the latissimus dorsi myocutaneous flap is an ideal treatment to severe craniomaxillofacial defects as it possesses the advantages of reliable blood supply, ability against infections, large size, concealed donor site, and functional restoration of sensation and movement. PMID- 15334931 TI - [Thoraco-abdomtinal flap for repairing the defects after separation of the thoraco-adbomino-conjoined twins]. AB - OBJECTIVE: To evaluate a technique for repairing a large thoraco-abdominal defect after the separation of the thoraco-abdomino-conjoined twins. METHODS: A 8 cm x 5 cm thoraco-abdominal flap on each side of the trunk of the conjoined twins was designed before the separation. The flap from the A-infant was used for repairing the thoraco-abdominal defect of the B-infant while the flap from the B-infant for the defect of the A-infant. RESULTS: Both of the defects after the separation of the conjoined twins were completely covered by the flaps. The flaps were survival very well with very good appearance. CONCLUSION: The above technique could be a good way for repairing the defects after the separation of the thoraco-abdominal conjoined twins. PMID- 15334932 TI - [Computer-assisted simulation and clinical application of custom-produced nose implant]. AB - OBJECTIVE: Surgery to reshape the nose with an implant has been a regular procedure for enhancing a patient's appearance and self-confidence. The purpose of this study was to establish a three-dimensional surgical simulation system. which can provide the patients with realistic prediction of their own postoperative appearance in computer and specifically produce a nose implant for an individual patient. METHODS: Preoperative CT data and image-processing techniques were employed to generate a three dimensional template of the papatient' s face. Three-dimensional morphing processes were then applied to predict and compare the outcome of plastic procedures on a patient-specific nose surface, according to the patient' s expectation. By comparing and revising the changes before and after simulation, digital prototypes of the nose implants could be generated. The templates to create custom-made silicone implants were than produced by a computer controlled device. RESULTS: Accurately regeneration of 3-D images and realistic operative simulations could be achieved with this system. The implants produced exactly conformed to the results of simulation. No curving and reshaping were needed during operating. The clinical results extremely matched with the simulations. CONCLUSION: The system enhances surgeon patient communication and facilitates preoperative planning. It is especially desirable for implant surgery with less guesswork of size, contour, and orientation of the implant. The best chance of optimal results could be achieved. PMID- 15334933 TI - [Metabolism of hyaluronic acid and extremity lymphedema]. AB - OBJECTIVE: To investigate the possible influence of the impairment of lymph fluid on the metabolism of hyaluronic acid (HA) in the lymphedematous skin tissue. METHODS: Tissue fluid was collected in lymphedematous limbs and the contralateral healthy limbs of 39 patients and HA content was measured with radioimmunoassay. The protein contents were also measured. RESULTS: The HA contents in interstitial fluid of lymphedematous limb were significantly (8 fold) higher than that of normal limb. The protein concentration in the tissue fluid did not show significant differences between lymphedema and those with normal tissue. CONCLUSION: The result suggests blockage of regional draining lymphatics may impairs breakdown of HA and the stagnation of HA in the limb may exert a deleterious effect on the interstitium. PMID- 15334934 TI - [Constricted ear therapy with free auricular composite grafts]. AB - OBJECTIVE: A simple and effective therapy for single side constricted ear. METHODS: Transplanting normal side free composite auricular grafts to constricted ear (15 patients and 15 sides), then lengthening the helix, exposing the scapha, correcting deformity. RESULTS: The 15 patients composite grafts all survived. The helix has been lengthened, the scapha exposed, the normal ear reduced, the constricted ear augmented and two sides ear have become symmetry. CONCLUSION: This method is simple and results are satisfied. PMID- 15334935 TI - [Orbicular muscular-mucous advancement flap for repairing the lower lip loss after lip cancer]. AB - OBJECTIVE: To explore a new technique for the treatment of lower lip defect after carcinoectomy. METHOD: Six lower lip defects (more than two third of the length of the lower lip) after the tumor resection were treated with an orbicular muscular-mucous advancement flap. RESULTS: All of the patients had achieved good results functionally and cosmetically with the following-ups from 6 months to 3 years. CONCLUSION: The above mentioned techique could be a simple, safe and effective method for repairing lower lip defect. PMID- 15334936 TI - [Autogenous costal cartilage with Medpor: a compound auricular framework applied to one-stage total ear reconstruction]. AB - OBJECTIVE: To introduce a compound external ear framework characterized by few complications, simplicity and very low rate of implant exposure or absorption. METHODS: A compound auricular framework, being composed of the ninth autogenous costal cartilage as ear rim and Medpor ear base, was used in one-stage total ear reconstruction. All patients were followed up postoperatively. RESULTS: Nine cases have been performed clinically using this compound framework and all of them achieved satisfactory efficacies. The reforged ears looked natural and achieved superior cosmesis, and none of them experienced ear rim fracture or implant exposure or distortion. CONCLUSION: This compound auricular framework has advantages of practicability, simplicity and minimal complications. It may be an ideal framework for the reconstruction of total external ear at present. PMID- 15334937 TI - [Collagen synthesis and expression of connective tissue growth factor in the cultured fibroblasts of human hypertrophic scar]. AB - OBJECTIVE: To explore the effect of connective tissue growth factor on the pathogenesis of human hypertrophic scar. METHODS: Normal skin and hypertrophic scar fibroblasts were cultured in vitro. The collagen synthesis of fibroblasts were measured by H3-proline incorporation method. The expression of connective tissue growth factor protein and mRNA of fibroblasts were detected with immunocytochemistry staining and reverse transcription polymerase chain reaction methods. RESULTS: Compared with normal skin fibroblast, the collagen synthesis and the expression of connective tissue growth factor protein and mRNA in the hypertrophic scar fibroblast was higher (P < 0.01). CONCLUSION: Connective tissue growth factor may play an important role in promoting the fibrotic process of hypertrophic scar. PMID- 15334938 TI - [Relationship of overexpression of angiogenesis factors and their receptors with invasive growth of keloid]. AB - OBJECTIVE: To study the relationship between invasive growth and the angiogenesis factors and their receptors in keloid. METHODS: Biopsies from 17 keloid (Ke) were divided into atrophy group (Ke-A, n = 9), proliferating group (Ke-P, n = 13), infiltrating group (Ke-I, n = 9), normal skin around Ke (Ke-N, n = 10) and normal skin (NS, n = 10). The histology, immunohistochemistry and computerized imaging analysis were used for the study. The levels of basic fibroblast growth factor (bFGF) and its receptor-Flg, vascular endothelial growth factor (VEGF) and VEGF/KDR complex (11B5), and platelet derived growth factor (PDGF-A) and its receptor-PDGFR-alpha, and alpha-smooth muscle actin (alpha-SMA) were determined in specimens with immuneohistochemical staining. RESULTS: In all 5 groups, bFGF, Flg, VEGF, 11B5, PDGF-A, and PDGFR-alpha were all expressed in fibroblasts (Fb), monocyte-phagocytes, vascular endothelial cells, adventitial cells, epidermal (cells and epithelial cells in appendage. The intensities of staining ranked as follows: Ke-I > Ke-N approximately equal to Ke-P > Ke-A approximately equal to NS, Flg > hFGF approximately equal to PDGFR-alpha > PDGF-A approximately equal to 11B5 > VEGF (P < 0.05 to approximately 0.01). 11B5 and VEGF were expressed (intensively in alpha-SMA positive myofibroblasts only in Ke-I group. The histological observation showed hyperplasia of endothelial cells and obliteration of microvessels. CONCLUSION: The invasive growth of keloid may be related to the overexpression of angiogenesis factors and their receptors. The abnormal expression of 11B5 in myofibroblasts may be one of the important factors associated with tumor-like growth feature in the invasive parts sites of keloid. The results suggest that inhibition of these biological activities would be of significance in clinical therapy. PMID- 15334939 TI - [Re-growth of induced-osseous tissue encapsulated in muscular fasciae for prefabrication of an osteo-musculo-cutaneous flap]. AB - OBJECTIVE: To improve the survival of mesenchymal stem cells (MSCs) for prefabrication of an osteo-musculo-cutaneous flap. METHODS: In a mouse model, the compound of MSCs and p(3HB-co-3HH) were embeded in the latissimus dorsi muscle as an experimental group and the muscle pocket of the buttock as the control. The examinations of the HE staining, hybridization in situ of osteonectin mRNA and Von kossa staining were used to evaluate the results. RESULTS: The expression of osteonectin mRNA and the Von kossa staining showed that the latissimus dorsi muscle group was superior to the control in 2 weeks and 4 weeks after the surgery in vivo. CONCLUSION: The results indicate that the above-mentioned technique may be a good alternative for the prefabrication of the osteo-musculo-cutaneous flap. PMID- 15334940 TI - [Mid-premaxillary sutural distraction osteogenesis for repair of alveolus cleft: an experimental study in dogs]. AB - OBJECTIVE: To explore a new technique for repair of alveolar cleft by sutural distraction osteogenesis. METHODS: Nine 8-weeks mongrel dogs were used in this study, three being in the control group, six in the experimental group. Alveolar cleft model was created surgically in all animals. Two weeks later, a U-shaped distractor made of Ni-Ti memory alloy wire was insterted into the premaxilla to distract the mid-premaxillary suture. When the premaxilla of the cleft side approached the ipsilateral maxilla, periosteoplasty of the alveolar cleft was performed. The distractor was removed at two weeks after periosteoplasty. The results were evaluated clinically, radiographically, morphologically and histologically. RESULTS: The cleft model in dogs was stable and similar to the human alveolar cleft. In experimental dogs, the premaxilla was moved gradually toward the maxilla so that the cleft was closed. The distracted mid-premaxillary suture showed a gradually widened traingle, with its tip being posterior. The density of the distracted traingle suture was increased gradually. Bony repair was achieved completely at the cleft three months post-periosteoplasy. The morphology of the mid-premaxillary suture was restored. CONCLUSION: The alveolar cleft could be repaired by the technique of mid-premaxilla suture distraction. PMID- 15334941 TI - [Influence of vacuum-assisted closure technique on expression of Bcl-2 and NGF/NGFmRNA during wound healing]. AB - OBJECTIVE: To explore the influence of vacuum-assisted closure technique (VAC) on expression of Bcl-2 and NGF during wound healing. METHODS: Eighty Sprague-Dawley rats were randomly divided into 4 groups with 20 rats of each. Group T was the experimental group; group C1, C2 and C3 were the control groups. In group T and group C1, capsaicin was injected subcutaneously to the back of the rats to destroy the sensory nerve. VAC was employed to the wound of the rats in group T and C2 three times a day at 80 mmHg negative pressure. In all the groups, tissue samples were taken from the wound edge and granulation at 1, 3, 6, 9 and 12 days after the injury. Immunohistochemistry and in situ hybridization were used to detect the expression of Bcl-2 and NGF/NGFmRNA in the samples. RESULTS: In group C2 and C3, the expression of Bcl-2 and NGF/NGFmRNA was obvious, which increased gradually and reached the peak at the 9th day. In the process of wound healing, the expression Bcl-2 and NGF/NGFmRNA was higher in the group C2 than in group C3 (P < 0.05). The expression Bcl-2 and NGF/NGFmRNA in group T and C1 was lower than group C2 and C3 (P < 0.05). CONCLUSION: The application of the vacuum-assisted closure technique during wound healing increases the expression of the apoptosic modulation related protein Bcl-2 and affects the expression of NGF/NGFmRNA, which may promote the wound healing process. PMID- 15334942 TI - [Biomass structure and quantitative relationship models of modules in clonal population of Puccinillia chinampoensis in Songnen plain]. AB - In this paper, quantitative analysis was conducted at module level on the biomass structure of modules in clonal population of Puccinillia chinampoensis, the relationship between modules' biomass and turf sizes, and the relationship between one module's biomass and another one's. Based on these, the corresponding models of these relationships were established. The results showed that the regularities on the biomass of all functional modules and their ratio were the same at earing stage and at vegetative stage after fruiting. There existed linear correlation between functional modules and turf sizes at earing stage, and exponential correlation at vegetative stage after fruiting. Among all quantitative relationship models, only the one between photosynthesis modules and supporting modules was linear function at earing stage and exponential correlation at vegetative stage after fruiting. The relationship models between any other two functional models were all exponential correlation at two stages. PMID- 15334943 TI - [Niche characteristics of plants on four environmental gradients in middle reaches of Tarim River]. AB - Tarim River is the longest continental river in China. Along its either bank, the natural vegetation belts are composed of arbors, shrubs and grasses. In this paper, the niche breadths and overlaps of nineteen species on four environmental gradients in the middle reaches of Tarim River were calculated and analyzed. The results showed that the nineteen species could be divided into three groups, based on their average niche breadths. Group one included Populus euphratica, Tamarix ramosissima, T. hispida, Lycium ruthernicum, Alhagi sparsifolia and Phragmites australis, which were constructive species in Tarim River and their niche breadths were greater. Group two included T. leptostachys, Aeluropus pungens, Poacynum hendersonii, Cynanchum sibiricum, Karelinia caspica, Inula salsoloides, Taraxacum sp., and Hexinia polydichotoma, and their niche breadths were smaller than that of group one. Group three included Halimodendron halodendron, Glycyrrhiza inflata, Calamagrostis pseudophragmites, Salsola sp., and Sophora alapecuroides. Based on the niche overlaps matrix, the niche overlaps of the plant species pairs were generally smaller on all the four environmental gradients, because of their probable niche shift or the demand for different environmental resources. Soil moisture and soil salinity were the important environmental factors limiting the plant distribution in the middle reaches of Tarim River. PMID- 15334944 TI - [Relationship between ephemeral plants distribution and soil moisture on longitudinal dune surface in Gurbantonggut desert]. AB - Ephemeral plant is a particular group of desert flora only distributed in northern Xinjiang, China. In this study, field surveys of vegetation and soil moisture were conducted in the southern area of the Gurbantonggut desert from March to August 2002. The species number, coverage and growth of ephemerals and soil moisture were measured in 7 quadrats at different geomorphic positions on the longitudinal dune surface. The results showed that soil moisture in a depth of 0-30 cm affected ephemeral plants distribution significantly. In early spring, the soil moisture was 4.62% on the intredune, 3.98% on the plinth, and 2.01% on the crest. Consequently, the average coverage of ephemeral plants reached 51.8%, 38.2% and 4.4%, respectively. The existence of ephemeral plants distribution influenced soil moisture as well. In later May, there was an increase of surface soil moisture from the interdune to the crest. The soil moisture was 1.00% on the intredune, 1.90% on the plinth, and 2.45% on the crest. PMID- 15334945 TI - [Analysis on age structure and dynamics of Kindonia uniflora populations]. AB - Kindonia uniflora is a perennial clone herbaceous plant, and also, a native endangered plant in China. This paper studied its age structure, life table and survivorship curve in different habitats in Taibai mountain area. The results indicated that the age structure and dynamics of K. uniflora populations in the Betula utilis forest at altitude 2500-2700 m, in the Abies fargesii forest at altitude 2700-2900 m, and in the Larix chinensis forest at altitude 2900-3100 m had the similar pattern and developing tendency. The number of younger ramets at 1-2 years old or older than 5 years was less, and the number of ramets at 3-5 years old was the highest in the age structures. The negative values of dx (dead number), qx (mortality rate) and Kx (Killing rate) in the life table showed the increasing rate of the population sizes during the age stage. The survivorship curve of K. uniflora populations in different habitats belonged to Deevey C after 3-5 years old. The mortality rate of populations during 5-10 years stage was higher, and was stable after 10 years old. As for the characters of asexual propagation and clone growth, the rhizomes of the populations were in humus of soil, and developed and expanded as guerilla line style. During growth season, only one leaf grew above ground at every inter-node, and the population growth and development were rarely influenced by external factors. The forest communities, such as Betula utilis, Abies fargesii and Larix chinensis forest, in which K. uniflora populations lived, were at middle or higher mountain, where there were rarely disturbance from human being. Therefore, the habitats for K. uniflora populations to live were relatively stable. As the altitude increased, the disturbances from human being became less, the density of K. uniflora populations increased, the life cycle expanded, the peak of population death delayed, and the population living strategy changed to adapt to the habitats. K. uniflora populations preferred to live in cool climate, thick humus of soil and under shaded forest, therefore, the habitat of communities of Abies fargesii, Larix chinensis forest at altitude 2700-3100 m were better than that in Betula utilis forest. The growth and development of K. uniflora populations depended on the habitats; therefore, the communities in which K. uniflora population lived should be conserved firstly. The in-situ conservation of the populations should be encouraged. PMID- 15334946 TI - [Molecular identification of Araceae-infecting viruses and their ecology]. AB - A filamentous virus and a spherical virus were detected from partial purification with natural-infected Araceae plants. The filamentous virus was confirmed as dasheen mosaic virus (DsMV) by RNA spot hybridization (RSH) and sequence determination, while the spherical virus was identified as a new strain of cucumber mosaic virus (CMV) by enzyme-linked immunosorbent assay and sequence determination of the 3' end genes of RNA3. CMV was firstly confirmed as one of the principal viruses infecting the Araceae plants under natural condition. According to the sequence similarity of coat protein, Araceae-infecting CMV was then decided belonging to CMV subgroup I, but independent from other isolates in this subgroup. By dot-RNA hybridization to DsMV and CMV probes with total RNA extracted from leaf tissue, field samples collected from some parts of southern China were detected for their natural infection. The infection ratio of field samples infected by DsMV and CMV were 73.3% and 46.7% for samples collected from Hainan Province, 100% and 38.5% for those from Hunan Province, 93.0% and 38.5% for that from Zhejiang Province, and 100% and no infection for samples from Shanghai, respectively. The infection of viruses on Araceae plants was obviously affected by such ecological factors as geological distribution, climate and host species. The occurrence of CMV was greatly influenced by the above factors, but DsMV always existed as the most commonly distributed virus. PMID- 15334947 TI - [Diversity of butterflies in Lianhua Mountain Nature Reserve of Dongguan City, Guangdong Province]. AB - This paper dealt with the diversity of butterflies collected from Lianhua Mountain Nature Reserve of Dongguan in April, June, September and December 2001. In this area, there were 50 species butterflies belonging to 9 families, 36 genera. Data analysis showed that the species richness and the highest diversity index were the highest in June, followed by in September, April and December, and the evenness index was the highest in April, then in September, June and December. The dominance index was the highest in December, then in September, June and April. At the level of families, Nymphalidae was the highest in richness, diversity [H'(GS), H'(G)] and evenness, the second was Papilionidae, and then was Hesperiidae. As for individual quantity, Danaidae was the most abundant, and the following were Pieridae and Papilionidea. For diversity index [H'G(S)], Papiliondae was the highest, and the next was Nymphalidae. PMID- 15334948 TI - [Pollen combination and paleo-environment of Momoge Lake since 1500 years before]. AB - Based on the pollen combination, 14C dating, and depositional facies analysis of Momoge profile in Zhenlai County, Jilin Province, three pollen belts were divided. The results showed that Momoge Lake was formed by blocked paleo-river course depression 1500 years ago, which included three development stages, i.e., the period when river converted into lake, the early period of lake development, and the steady period. Its corresponding plant types were Artemisia-Chenopodium grassland, Artemisia-Chenopodium grassland accompanied by Betula, and meadow grassland, and its paleo-climate was dry-cold, warm-dry, and then cool-wet, and there was a warm-dry tendency in recent fifty years. PMID- 15334949 TI - [Ecological management model of agriculture-pasture ecotone based on the theory of energy and material flow--a case study in Houshan dryland area of Inner Mongolia]. AB - The degradation of ecological environment in the agriculture-pasture ecotone in northern China has been paid more attentions. Based on our many years' research and under the guide of energy and material flow theory, this paper put forward an ecological management model, with a hill as the basic cell and according to the natural, social and economic characters of Houshan dryland farming area inside the north agriculture-pasture ecotone. The input and output of three models, i.e., the traditional along-slope-tillage model, the artificial grassland model and the ecological management model, were observed and recorded in detail in 1999. Energy and material flow analysis based on field test showed that compared with traditional model, ecological management model could increase solar use efficiency by 8.3%, energy output by 8.7%, energy conversion efficiency by 19.4%, N output by 26.5%, N conversion efficiency by 57.1%, P output by 12.1%, P conversion efficiency by 45.0%, and water use efficiency by 17.7%. Among the models, artificial grassland model had the lowest solar use efficiency, energy output and energy conversion efficiency; while the ecological management model had the most outputs and benefits, was the best model with high economic effect, and increased economic benefits by 16.1%, compared with the traditional model. PMID- 15334950 TI - [Evaluation of agricultural net primary productivity in Yijinhuoluo County, Inner Mongolia]. AB - An agricultural net primary productivity model considering both climatic and fertilizer factors was presented, with referencing over 40 years (1959-1998) climatic and agricultural data from Yijinhuoluo County. Based on the scenarios of climate, population and human activities (mainly fertilization) in the next 30 years as well as the demand of local people for food, the changes of the crop area were evaluated. As a result, 622-5948 hm2 could be converted into grassland and forest land from now to the year of 2010, amounting to 3%-31% of the average area between 1990 and 1998; from 2010 to 2020, the converted area should be 3263 8164 hm2, amounting to 17%-42%. In the next 10 years, the area of cropland would rise slightly because of the limitation of increasing yield and population. PMID- 15334951 TI - [Effect of land-use change on soil habitat in north Hebei plateau during last 50 years]. AB - The change of soil habitat in an area depends on the land-use and cover change in the area to a great extent. Analysis of soil data in north Hebei plateau during last 50 years confirmed that the land-use change, i.e., the disorder reclamation of a large stretch of natural grassland into cultivated land and the extensive farming, had a series of negative effects on the soil habitat, and made the soil nutrients decreased. During last 50 years, the total and available amount of soil organic matter, N, P, and K decreased by 36.90%, 67.77%, 44.44%, 34.15%, 32.22%, 58.90% and 35.29%, respectively, and the granulometric contents of sand, silt and clay also varied by 18.24%, 19.55%, and 12.29%, respectively, which revealed that in order to ensure the land quality in the regionally important but vulnerable habitat area against the effect of land-use change and its possible degradation with time and to be in sustained dynamic equilibrium, it is necessary to take reasonable fertilization and reliable technical measures. PMID- 15334952 TI - [Impact of land-use change on soil carbon storage]. AB - Through comparing the concentration and inventory of soil organic carbon (SOC) and its distribution in soil profiles under cropland, rangeland, natural secondary forest (brushwood, natural secondary forest dominated by Querces liaotungensis or Populus davidiana) and larch plantations (13, 18 and 25 years old Larix principisrupprechtil), this paper studied the effect of land use change from natural secondary forest to cropland or rangeland as well as from cropland or rangeland to plantation on SOC storage in the Liupan mountain forest zone. The results showed that the concentration of SOC in 0-110 cm soil layer under cropland and rangeland was 54% and 27% lower than that under natural secondary forest, respectively. The difference of SOC concentration between natural secondary forest and cropland or rangeland was greater in 0-50 cm than in 50-110 cm soil layer, while that between larch plantations and cropland or rangeland was greater in 0-40 cm than in 40-110 cm soil layer. The inventory of SOC in 0-110 cm soil layer under cropland and rangeland was respectively 35% and 14% lower than that under natural secondary forest, while 23% lower under cropland and 4% higher under rangeland than that under larch plantations. The difference of SOC inventory between natural secondary forest and cropland or rangeland was greater in 0-50 cm than in 50-110 cm soil layer, while that between plantations and cropland or rangeland was greater in 0-30 cm than in 30-110 cm soil layer. The decreasing magnitude of SOC storage with soil profile depth under natural secondary forest or larch plantations was greater than that under cropland or rangeland. The above-mentioned facts resulted from the changes of SOC input or output and the distribution of roots in soil. The results indicated that the SOC concentration and inventory would decline (mainly in 0-50 cm soil layer) after converting from natural secondary forest to cropland or rangeland, but increase (mainly in 0-30 cm soil layer) following afforestation on cropland. The SOC concentration would increase but its inventory would not change following afforestation on rangeland, and the distribution of the SOC concentration or inventory in soil profile would change with the change of land use in Liupan mountain forest zone. PMID- 15334953 TI - [Community structure and seasonal change of soil micro-arthropodes in the Lower Reaches of Liaohe River Plain under different land utilization]. AB - The data on the soil micro-arthropodes under four land utilization types (fallow, forest, upland field and paddy) in the Lower Reaches of Liaohe River Plain were collected in a period from October 2000 to October 2001. Using the community parameters of population density, group richness, diversity index and evenness, the community structure and its seasonal changes were described. There were 12 groups of soil micro-arthropodes in this region, and of the groups, Collembola and Acarina were dominant, and Diptera, Coleoptera and Hymenoptera were often seen in fallow, forest and upland field, while Collembola, Acarina and Diptera were dominant in paddy. Both land utilization and seasonal change influenced the population density, group richness and diversity index of soil micro-arthropodes. The vertical distribution in both density and group number of arthropods in soil was in the order of surface > middle > bottom. PMID- 15334954 TI - [Effect of herbicide quinclorac on microbic populations in a paddy soil]. AB - The potential effect of herbicide quinclorac on culturable microorganisms was investigated in a flooded paddy soil added with 0.33, 0.67, 1.00, 1.33, and 2.00 microg quinclorac x g(-1) dried soil, by a 10-fold serial dilution plate technique for total soil aerobic bacteria, actinomycetes and fungi, by three-tube anaerobic most-probable-number (MPN) method with anaerobic liquid enrichment media for anaerobic fermentative bacteria (AFB), denitrifying bacteria (DNB) and hydrogen-producing acetogenic bacteria (HPAB), and by the rolling tube method in triplicate for methanogenic bacteria (MB) and nitrogen-fixing bacteria (NFB). The results showed that there were some differences in the aerobic heterotrophic bacteria, AFB and DNB between soils supplemented with quinclorac and non quinclorac at the early stage of incubation, but none of them was persistent. The numbers of fungi and DNB were increased in soil samples treated with lower than 1.33 microg quinclorac x g(-1) dried soil, while the CFU of fungi and HPAB were decreased in soil samples treated by higher than 1.33 microg quinclorac x g(-1) dried soil. The population of actinomycete declined in negative proportion to the concentrations of quinclorac applied after 4 days, and the application of quinclorac greatly stimulated the growth of AFB and NFB. MB was more sensitive to quinclorac than the others, but its number in the soil samples with lower concentrations of quinclorac was nearly equal to that in the control on the 33rd d. It could be concluded that quinclorac was safe to the soil microorganisms when applied at normal concentrations (0.67 microg x g(-1) dried soil). PMID- 15334955 TI - [Spatial distribution pattern of Chilo suppressalis analyzed by classical method and geostatistics]. AB - Two original samples of Chilo suppressalis and their grid, random and sequence samples were analyzed by classical method and geostatistics to characterize the spatial distribution pattern of C. suppressalis. The limitations of spatial distribution analysis with classical method, especially influenced by the original position of grid, were summarized rather completely. On the contrary, geostatistics characterized well the spatial distribution pattern, congregation intensity and spatial heterogeneity of C. suppressalis. According to geostatistics, the population was up to Poisson distribution in low density. As for higher density population, its distribution was up to aggregative, and the aggregation intensity and dependence range were 0.1056 and 193 cm, respectively. Spatial heterogeneity was also found in the higher density population. Its spatial correlativity in line direction was more closely than that in row direction, and the dependence ranges in line and row direction were 115 and 264 cm, respectively. PMID- 15334956 TI - [Time-concentration-mortality modeling for responses of Myzus persicae to low sublethal sprays of imidacloprid]. PMID- 15334957 TI - [Efficacy of emulsifiable formulations of two entomopathogenic fungi against small green leafhoppers on tea plant]. AB - A field trial was conducted to test the efficacy of emulsifiable formulations of Beauveria bassiana (Bb) and Paecilomyces fumosoroseus (Pf) conidia in controlling small green leafhoppers (Empoasca spp.) on tea plant in a hillside tea garden located in Shuichang, Zhejiang during mid-summer 2002. Both Bb and Pf formulations contained 10(10) conidia x ml(-1). Adding 3% of imidacloprid 10% WP to each fungal formulation (W/V) resulted in two other formulations, i.e., Bb imidacloprid mixture (BbIM) and Pf-imidacloprid mixture (PfIM). Besides the four formulations, a mineral oil-based liquid used to formulate the fungal conidia and containing 3% of imidacloprid 10% WP (OBLI) was also tested, and water spray was used as control (CK). Each of the treatments included three 60-m2 plots (replicates), which were sprayed twice with a 500-fold aqueous dilution of the corresponding formulation or preparation at a 12-d interval. Based on the leafhopper densities estimated by sampling in all plots at 3- or 4-d intervals after the first spray, the spray of BbIM or PfIM could better control the leafhoppers than the spray of Bb or Pf formulation. The maximal efficacy relative to CK reached 83.4% and 71.3% in the BbIM and PfIM treatments, respectively. Estimates of field efficacy obtained during the 25-d period after the first spray was 66.8% for BbIM, 62.1% for PfIM, 50.3% for OBLI, 49.5% for Bb, and 19.0% for Pf, respectively. A discussion was also given on the effect of local weather pattern and leafhopper population age structure on the results of the field trial, and on the application techniques to enhance the efficacy of mycoinsecticides against Empoasca species on tea plant. PMID- 15334958 TI - [Attraction effect of main volatile components from tea shoots and flowers on Sphaerophoria menthastri (Diptera: Syrphidae) and Chrysopa septempunctata (Neuroptera: Chrysopidae)]. AB - The Y-olfactometer bioassays showed that the volatiles from tea aphids and tea aphid-damaged tea shoots (PHC) strongly attracted Sphaerophoria menthastri and Chrysopa septempunctata. At the dosage of 10(-4) g x ml(-1), S. menthastri selected geraniol and methyl salicylate from PHC, n-octanol from intact tea shoots (ITS), nerol from tea flowers (P < 0.01), and (E)-2-hexen-1-ol, E-2 hexenal, 1,3,6-octatriene-3,7-dimethyl and linalool from PHC (P < 0.05), while C. septempunctata selected n-octanol and hexanal from ITS (P < 0.05). At the dosage of 10(-2) g x ml(-1), S. menthastri selected hexanal, nerol and benzaldehyde from PHC (P < 0.05), while C. septempunctata selected hexanal, benzaldehyde, geraniol and (E)-2-hexen-1-ol (P < 0.05). The experimental results showed that besides aldehydes and methyl salicylate, geraniol, n-octanol and (E)-2-hexen-1-ol were significantly attractive to the natural enemies. The selective responses of natural enemies were restricted by thresholds. Both dosages of hexanal significantly attracted C. septempunctata. Nerol might be an infochemicals seducing S. menthastri to orient to tea flowers. PMID- 15334959 TI - [Comparison of habitats and seasonally differentiated distribution patterns of fig wasp populations associated with Ficus racemosa in Xishuangbanna]. AB - The population distribution patterns of fig wasps associated with Ficus racemosa in Xishuangbanna were studied by using several indexes. The results indicated that the distribution patterns of 6 fig wasp species were all clump. The aggregative intensity of pollinating wasp (Ceratosolen fusciceps) population at primeval rain forest was nearly equal to that at seriously disturbed site, whereas they were both much higher than that at moderately disturbed site. In the meanwhile, the population aggregative intensity for the same wasp species was higher in rainy season than in dry-hot season, and the lowest was occurred in foggy-cool season. Non-pollinating wasp (Platyneura testacea, Platyneura mayri, Platyneura agraensis, Apocrypta westwoodi, Apocrypta sp.) population and each of their sex group were varied significantly in aggregative intensity, responding to the habitat change and seasonal alteration. The protection of original habitat should be focused on the protection of the mutualism of Ficus racemosa and fig wasps. PMID- 15334960 TI - [A dynamic knowledge model for designing suitable sowing date of rice]. AB - Based on the summarization, induction and extraction of dynamic relationships between growth characteristics, variety type, farming system and ecological conditions via reading literature and consulting experts, a knowledge model for designing the suitable sowing date of rice under varied environments was developed through knowledge engineering and system analysis methods. The model could precisely quantify the suitable sowing date under different environments and production systems. Case studies for the model with the data sets of daily weather conditions, typical varieties, seedling nursery methods and farming systems at 7 different eco-sites indicated a good performance of the model system in decision-making and its wide applicability, and the model could help farmers to use the resources such as sunshine and water effectively. PMID- 15334961 TI - [Effect of rice-duck mutualism on nutrition ecology of paddy field and rice quality]. AB - The production of nuisanceless rice was conducted by the technique of rice-duck mutualism without applying any pesticide and chemical fertilizer. The results showed that compared to the control, the effect of eliminating weeds was over 99.4%, and the cardinal numbers of plant diseases and insect pests reduced obviously. Nutrients and dissolved oxygen in paddy water and rapidly available nutrients in soil increased, but after maturing stage, compared to basal fertility, rapidly available P and K decreased. The N, P, and K absorption amounts of rice plant, rice yield, setting panicles rate, filled grains and ripened rate increased. The milling, appearance, nutritional, cooking and eating quality of rice improved, especially in reducing chalkiness. The comprehensive benefits of paddy field enhanced obviously. PMID- 15334962 TI - [Effects of nutrient and water regimes on lodging resistance of rice]. AB - A field experiment was conducted to study the effects of nutrient and water regimes on lodging resistance of rice plant by measuring and comparing the morphological and physical traits and the K2O and SiO2 contents of rice basal stems. The results indicated that a combined application of chemical fertilizers and organic materials, especially and rice straw (CS), could markedly increase the diameter, wall thickness and weight of basal stems, but reduce plant height, which resulted in the effective increase of snapping resistance (RS) and marked decrease of lodging index (LI). The effectiveness of the combined application of chemical fertilizers and rice straw (CS) on the increase of lodging resistance of rice plant was greater in alternate wetting and drying (AWD) and controlled irrigation (DRA), compared to continuous waterlogging (CWL). In addition, the combined application of chemical fertilizers and organic sources, especially with rice straw (CS), greatly increased the K2O and SiO2 contents in rice basal stems, especially in alternate wetting and drying (AWD) and controlled irrigation (DRA). Correlation analysis showed that the wall thickness, weight and snapping resistance of basal stems were significantly positively correlated to the K2O and SiO2 contents in rice basal stems (P<0.05 or P<0.01) in alternate wetting and drying (AWD) and controlled irrigation (DRA). PMID- 15334963 TI - [Effects of lanthanum on root membrane permeability and nutrient contents in root exudate of rice]. AB - This study showed that when lanthanum was in lower concentration (< or = 50 microg x ml(-1)), the membrane permeability of rice root was stabilized by decreasing the electrolyte exosmosis, and the contents of K+, Ca2+, P and H+ in root exudate were lower than the control; while the concentration of La3+ was in the range of 100-400 microg x ml(-1), the electrolyte exosmosis was reduced in the first 6 hours, and then significantly increased. When the concentration of La3+ was over 500 microg x ml(-1), the electrolyte exosmosis was continuously enlarged because of the destruction of the membrane. Under this condition, K+, Ca2+, P and H+ in root exudate were higher than the control. It could be concluded that the concentration of 50 microg x ml(-1) La3+ was safe to rice growing. PMID- 15334964 TI - [Effects of nitrogen forms on roots and N fertilizer efficiency of different wheat cultivars with specialized end-uses]. AB - A pot experiment showed that under NH2-N treatment, strong gluten wheat Yumai 34 had the highest root biomass, root activity, N use efficiency, N harvest index and grain protein content, but all of these were the lowest under NO3(-)-N treatment. For the medium gluten wheat Yumai 49, the items measured were the highest under NH4+-N treatment, but not the same trend under NO3(-)-N and NH2-N treatments. For the weak gluten wheat Yumai 50, NH4+-N treatment was the best for its top quality and specialized end-uses. There was a significant relationship between grain protein content and N harvest index. PMID- 15334965 TI - [Effect of low temperature stress on chilling tolerance and protective system against active oxygen of grafted watermelon]. AB - This paper studied the chilling tolerance and the activity of protective system against active oxygen of own-rooted and grafted watermelon seedlings. The results showed that under low temperature stress, the chilling tolerance of the watermelon seedlings grafted on figleaf was higher than that of the seedlings grafted on Chaofeng F1, and that of the own-rooted seedlings was the lowest. The decrease of chlorophyll, ascorbate and GSH contents and superoxide dismutase, ascorbate peroxidase and dehydroascorbate reductase activities as well as the increase of MDA content in the leaves of both own-rooted and grafted watermelon seedlings indicated an injury on the protective system against active oxygen, and induced lipid peroxidation under low temperature stress. However, the activity of protective system against active oxygen was higher in grafted watermelon seedlings than in own-rooted watermelon seedlings. The activity of protective system against active oxygen in figleaf gourd-grafted watermelon seedlings with stronger chilling tolerance was higher than that of Chaofeng F1-grafted watermelon seedlings with weaker chilling tolerance. It could be concluded that the enhancement of chilling tolerance in grafted watermelon seedlings was related to the increase of antioxidants and antioxidative enzyme activities. PMID- 15334966 TI - [Physiological and ecological effects of potassium on expansion of crocus corm]. AB - The corm size of Crocus sativus, a well-known medicine, is a key limiting factor for its stigma harvest. A water cultural experiment with four different potassium (K) concentrations showed that the K content, chlorophyll content, relative ATP content and net photosynthetic rate in C. sativus leaves increased with improving K concentration, which enhanced the function of leaves to be metabolic source. On the other hand, the K absorbing ability and the contents of soluble sugars, vitamin C (Vc) and protein in new-born corms were increased with improving K concentration, which also strengthened the function of new-born corms to be metabolic sink. This study also showed that K could promote the new-born corm growth rate of C. sativus through strengthening the function of "source" and "sink". PMID- 15334967 TI - [Interactive effect of N and Ca on N uptake by tomato]. AB - To study the effect of fertilizer N and Ca on tomato plant, a pot experiment was conducted with meadow soil in Shenyang. The results showed that under the experimental condition, the N contents in stems, leaves and fruits decreased gradually along with the growth stages. For all treatments, the NO3(-)-N content in fruits, which was within the raw eatable standard, had a positive correlation to N fertilization level. Adequate application of Ca was useful to control the NO3(-)-N content. The response of N contents in stems, leaves and fruits to fertilizer N and Ca was in line with the law of diminishing marginal returns. From flowering stage to fruit expanding stage, the response of fertilizer N was most activated. The nitrate reductase activity in leaves might be greatly affected by fertilizer N, while adequate Ca fertilization could decrease it. A combined application of N and Ca could promote the nitrate reductase activity. Both soil NO3(-)-N content and electric conductivity (EC) were increased with the increasing amount of applied fertilizer N. PMID- 15334968 TI - [Emergy analysis on an added loop in Spartina alterniflora ecological engineering]. AB - Based on the principle of multi-grade utilization of resources to get ecological, economic and social benefits of ecological engineering, this paper designed an added loop, following the Spartina alterniflora Ecological Engineering (SAEE). All the added loop design included SAEE, and the capsule was named SAEEC. In the added loop design, the Biological Mineral Liquid (BML) was made into antihyperlipidemia capsules, of which, the total flavonoids added up to 9.58 mg x g(-1). Emergy analysis method was applied to evaluate the SAEEC project. Compared with SAEE, the added loop design increased emergy investment ratio (EIR) by 1.37 fold, net economic benefit of the SAEEC by 2.13 fold, economic yield/input ratio by 1.46 fold, net emergy yield (NEY) by 3.18 fold, and net emergy yield ratio (EYR) by 2.20 fold, showing its more efficiency. PMID- 15334969 TI - [Effects of different temperatures on the growth and energy budget of Chinese shrimp, Fenneropenaeus chinensis]. AB - The effect of temperature on the growth and energy budget of Chinese shrimp Fenneropenaeus chinensis was studied at 18, 22, 25, 28, 31 and 34 degrees C. The results showed that its specific growth rate in terms of body weight and energy accumulation increased with temperature between 18 degrees C and 31 degrees C, and then decreased significantly at 34 degrees C. The similar trend was observed in food consumption and apparent digestion rate. The food conversion efficiency in weight and energy were 28.99%-53.09% and 15.70%-7.24%, respectively, which decreased with increasing temperature. The optimum temperature for the growth of shrimp was calculated from the relationship of SGR, which was 29.7 degrees C in this study. The energy budget of shrimp showed that the energy assimilated from food decreased with increasing temperature, while that spent in metabolism increased with increasing temperature. The results indicated that the high growth rate of Chinese shrimp at suitable temperature mainly resulted from the significant increase of food consumption and apparent digestion rate at corresponding temperature regimes. PMID- 15334970 TI - [Spatial heterogeneity of demersal fish in East China Sea]. AB - Quantitative analysis for the spatial distribution of fish is one of the important methods in fishery or fish ecology research. In this paper, the indexes Geary c and Moran I for the density distribution of demersal fish were calculated, and the semivariograms were drawn. The values of Geary c and Moran I were 0.38 and 0.52, respectively, and the C0/C0 + C was 59.9%, which meant that the distribution had a medium spatial autocorrelation with anisotropy, and the heterogeneity caused by random was a little higher than that caused by spatial autocorrelative process. The annual fluctuation of density was caused by the spatial autocorrelation and the random, because the density was significantly positively correlated with the values of C, C0 and C + C0, respectively. PMID- 15334971 TI - [Characters of greening tree species in heavy metal pollution protection in Shanghai]. AB - In this paper, the stream banks nearby Qibao town and the factory area of Shanghai Baoshan Steel Company were selected as the typical areas contaminated by heavy metals. The polluted status was investigated by measuring the heavy metal concentrations of the sampled soils. The results showed that the heavy metal concentrations in the soils of stream banks were a little higher than the control, but obviously higher in the factory area of Shanghai Baoshan Steel Company. The growth status of the greening trees was recorded, and their heavy metal concentrations were measured by ICP. According to the research results and historic data, the excellent greening tree species mainly applied in polluted factory area were Viburnum awabuki, Lagerstroemia indica, Hibiscus mutabilis, Ligustrum lucidum and Sabina chinensis, which could grow well on contaminated soil, and accumulate high concentrations of heavy metal elements. The other tree species such as Distylium racemosum, Nerium indicum, and Photinia serrulata might be also available in greening for heavy metal pollution protection. PMID- 15334972 TI - [Relationship between urban green-land landscape patterns and air pollution in the central district of Yichang city]. AB - In this paper, four types of landscape structures and their green-land landscape patterns, including the landscapes of dominant green-land patch pattern, even green-land patch pattern, dominant wooded corridor pattern and building or concrete covering pattern (control) in the central district of Yichang city in Hubei Province, were analyzed respectively on the basis of landscape ecological theory. The atmospheric noise and the contents of SO2, NOx and total suspension particle (TSP) of the landscapes were monitored respectively by comparative method. The results showed that the landscape of building or concrete covering pattern (control) was mostly composed of woodless corridors and building or concrete covering patches with the tiptop green-land fragmentation index (18.125 3 ind x hm(-2)) and only had 1.00% green-land coverage, which had a relatively higher atmospheric noise and the highest TSP content in the landscape. The landscape of dominant green-land patch pattern had the highest green-land coverage (up to 43.59%) dominated by great green-land patches and the least fragmentation index (0.453 9 ind x hm(-2)), in which, the atmospheric noise weakened by 28.12% and the TSP content reduced significantly by 86.42%, comparing to the control. The landscape of even green-land patch pattern had the relatively lower green-land coverage (11.34%) and fragmentation index (2.751 1 ind x hm( 2)), which was mainly composed of the middle or small green-land patches and wooded corridors with a regular distribution. In the landscape, the TSP content reduced obviously by 46.62% of the control, while the effect of dust retention was only 53.95% of that in the landscape of dominant green-land patch pattern. In the landscape of dominant wooded corridor pattern, which was a traffic center and turned into a main pollution resource in the city, there were a relatively higher green-land fragmentation index (6.870 0 ind x hm(-2)) and the highest wooded corridor density (0.844 3 hm x hm(-2)) with only 6.13% green-land coverage, and the atmospheric noise and the TSP and NOx content increased by 21.47%, 5.08% and 9.06%, respectively, comparing to control. It was obvious that the greater the average area of the green-land patch and the lower the fragmentation index of green-land patches, the more effective the green-land on purifying air pollution. PMID- 15334974 TI - [Current situation and some intending problems of landscape spatial dynamic model]. AB - Along with the rapid development of remote sensing, geographic information system, computer and other supporting technologies, model approach has become the major method of landscape dynamic study. Based on literature review, this paper analyzed the current situation and some intending problems of the development of landscape spatial dynamic model. Three kinds of models, i. e., stochastic landscape model, neighborhood rule model and landscape process model (including percolation model, individual-based landscape model and spatial ecosystem model) were mainly introduced, and their development status, existing problems and related improvement work were analyzed. The development of model test technologies, such as verification analysis, validation analysis, and sensitivity analysis, were also analyzed. Finally, this paper summed up six major problems that should be dealt with in the future development of the landscape spatial dynamic models. These problems included model arithmetic optimization, scale conversion, model complication and predigesting, model test and assessment, bringing forward of general model, and integration of the traditional model with social-economic factors. PMID- 15334973 TI - [Effect of the combination of cadmium and UV-B radiation on soybean growth]. AB - This paper studied the characteristics of soybean growth under the stress of Cd, UV-B radiation and their combination. The results showed that UV-B radiation gave a more pronounced inhibition on soybean growth than cadmium. Under UV-B radiation, the internode differentiation and elongation were inhibited, the internode and plant height were reduced, and the activities of SOD and POD had an apparent increase. Cd2+ could antagonize the effect of UV-B radiation on POD activity, and inhibit indole-3-acetic acid (IAA) oxidase activity. A significant decrease of POD and IAA oxidase activities in soybean was detected when exposed to combined UV-B and Cd2+, compared with UV-B radiation only. The decrease of POD and IAA oxidase activities was related to the increase of IAA contents and photosynthetic activity, which might account for the increases of biomass and plant height. Moreover, the combined stress enhanced the inhibition of root growth, and significantly decreased root length. The IAA oxidase and POD activities and photosynthesis rate might be the crucial factors determining the pattern of soybean growth under the stress of Cd2+ in combination with UV-B radiation. PMID- 15334975 TI - [Chemical mechanism of exotic weed invasion]. AB - The ecological and economic impacts of exotic invasive weeds have been followed with interest. A successful invasion of exotic weeds depends not only on their bio-ecological traits, but also on some specific mechanisms of invasion. The clarification of the invasive mechanism of exotic weeds will be beneficial to predict and manage on them. In the process of establishment, reproduction and spreading, an exotic species has to become dominant in the interactions with native ones at new habitats, and then invades successfully. Among invasive mechanisms, the chemical interaction between exotic and native weeds should not be neglected. Previous studies showed that allelopathy plays an important role in exotic weed invasion. In fact, plants are able to defend or inhibit animals, plants and microbes from new habitats by using some specific chemicals. The chemical mechanism of exotic weed invasion might involve each aspect of plant chemical ecology. Therefore, chemical ecological characteristics of exotic weeds should be regarded as one of the most significant factors on predicting invasive species. The studies on chemical mechanism of exotic weed invasion will be one of the important aspects in future. PMID- 15334976 TI - [Research advances in mechanism of high phosphorus use efficiency of plants]. AB - Phosphorus deficiency is one of the main factors influencing agricultural and forestry productions. Fertilization and soil improvement are the major measures to meet the demand of phosphorus for crops in traditional agriculture and forestry management. Recently, the plants with high phosphorus use efficiency have been discovered to replace the traditional measures to improve phosphorus use efficiency of crops. This paper reviewed the research advances in the morphological, physiological and genetics mechanisms of plants with high phosphorus use efficiency. There were three mechanisms for the plants with high phosphorus use efficiency to grow under phosphorus stress: (1) under low phosphorus stress, the root morphology would change (root system grew fast, root axes became small, the number and density of lateral root increased) and more photosynthesis products would transport from the crown to the root, (2) under low phosphorus stress, plant root exudation increased, mycorrhizae invaded into root system, the feature of root absorption kinetics changed, and the internal phosphorus cycling of plant reinforced to tolerate phosphorus deficiency, and (3) under long selection stress of low phosphorus, some plants would form the genetic properties of phosphorus nutrition that could exploit the hardly soluble phosphorus in the soil. PMID- 15334977 TI - [Concepts and relative analytical techniques of soil organic matter]. AB - The research of soil organic matter (SOM) has been highlighted in soil science. In the past 50 years, new perspectives in the relationship between SOM and sustainability of atmosphere and biomosphere, and strong motivation to find a vivid index for soil quality variation induced the transformation in concepts and analytical techniques of SOM: the curiousness to humic substances faded off since they were dull to anthropogenic activities, and interests were focused on the light fraction of organic matter (LFOM), organic carbon associated with different mineral particles in size, particulate and intra-particulate organic matter (POM and iPOM), water soluble organic matter (WSOM), and microbial biomass carbon (MB C). The relative fractionation procedures have been developed, and the main research activities on SOM are transformed from the products of microorganisms (humus) to the organic matter comprised in plant residues at their various decomposition stages and the organic carbon in microorganisms, since they are biologically active and immediately respond to soil cultivation and tillage, crop rotation, and fertilizer application, etc. PMID- 15334978 TI - [Research advances in ecophysiological effects of starvation on crustacean]. AB - As a major environmental stress, starvation greatly and extensively affects crustacean in ecophysiological aspects. In this paper, the existing information on the ecophysiological responses of crustacean to starvation, including development and survival, metabolism, digestive gland and enzyme, body composition, and recovery growth was summarized, and on the basis of these, the future research directions, i. e., the compensatory growth and its mechanism, and the relationship between starvation and immunocompetence, were proposed. PMID- 15334979 TI - [Influence of different NH4+/NO3- ratios on nitrogen metabolism of cotton]. AB - The influence of different NH4+/NO3- ratios on nitrogen metabolism of cotton was studied under controlled hydroponics. The results showed that compared with single nitrate nutrition, solutions with 25/75, 50/50, 75/25 and 100/0 of NH4+/NO3- significantly increased the soluble protein accumulation in leaves and roots of cotton, and the maximum content of soluble protein in leaves and roots appeared respectively in the solution with 50/50 and 75/25 of NH4+/NO3-. The soluble protein content in roots was increased with the increase of NH4+ percentage, but was slightly less in the solution of 100/0 than 75/25, which was probably related to the excess NH4+ limiting boot metabolism. With the increase of NH4+ percentage, the nitrate content in petiole and the nitrate reductase activity in functional blade declined, but ammoniac nitrogen content increased in every organ of cotton. These results showed that foreign nitrogen affected the nitrogen metabolism of cotton in a different way, and the nitrogen absorption by cotton was probably related to different forms of foreign nitrogen. PMID- 15334980 TI - [Interactive system of Delphastus catalinae and its competitor: interspecies competition]. AB - Delphastus catalinae and Serangium japonicum are the important natural enemies of Bemisia tabaci, of which, the former is exotic, and the latter is indigenous. In this paper, the influence of interspecies competition on the population growth of D. catalinae and S. japonicum was studied under conditions of single and mixed rearing, and Lotka-Volterra equation was used to simulate the competition when more and less preys provided for the two ladybirds. The results showed that the population growth of D. catalinae was greater than that of S. japonicum when single rearing, and the population growth of S. japonicum was greater than that of D. catalinae when mixed rearing. In the competition system of the two ladybirds, S. japonicum was dominant, while D. catalinae could exist simultaneously. PMID- 15334981 TI - [Effect of different ecological factors on ricefield eel (Monopterus albus) hatching rate]. AB - This paper studied the effects, of some ecological factors (temperature, pH, hatching method and ammonia) on the embryonic development of ricefield eel. The results indicated that the optimum hatching temperature of ricefield eel was 25 28 degrees C, and no significant difference was found about the effects of pH within the range of pH 5.5-9.5 on the hatching rate of oosperm. Dripping water incubation had a higher hatching rate of oosperm than staticing water incubation, and the hatching rate of oosperm decreased with increasing ammonia content. The present study provided an available suggestion for the all-artificial and half artificial breeding of ricefield eel. PMID- 15334982 TI - Psychotherapy, revisited. PMID- 15334983 TI - Cognitive-behavioral therapy for schizophrenia: a review. AB - Cognitive-behavioral therapy (CBT) has a proven role as an adjunct to antipsychotic medication and remediative approaches such as social skills training in the management of residual symptoms of chronic schizophrenia. Positive symptoms, depression, and overall symptoms appear to be viable treatment targets for CBT with a less pronounced effect on negative symptoms. The effect size at end of therapy is strong, with durability at short-term follow up. CBT can be used safely in patients with schizophrenia, and caregivers can help with homework exercises. There is also evidence that psychiatric nurses in the community can use CBT effectively with this patient group under supervision. CBT can be combined with family therapy and assertive community treatment programs targeted to reduce relapse. CBT improves the coping of patients with schizophrenia through improved adherence and symptom management. CBT techniques include development of trust, normalizing, coping strategy enhancement, reality testing, and work with dysfunctional affective and behavioral reactions to psychotic symptoms. An enhanced response to CBT would be expected when given with low dose cognitively enhancing atypical antipsychotic medication. PMID- 15334984 TI - Treatment of cognitive dysfunction in psychiatric disorders. AB - Most people with severe and persistent psychiatric disorders have significant cognitive deficits. Both schizophrenia and affective disorders with psychosis are associated with problems in attention, memory, and executive functioning. These deficits are predictive of lower work status, impairments in social skills, and poorer response to psychosocial skills training. Cognitive impairment makes it difficult for individuals to benefit from focused skills interventions that they are offered. This article describes behavioral treatments available to address cognitive dysfunction in individuals with psychiatric disorders. The evidence for efficacy is reviewed and recommendations are made for ways to begin addressing cognitive impairment in mainstream psychiatric practice. PMID- 15334985 TI - Cognitive-behavioral treatment for PTSD among people with severe mental illness: a proposed treatment model. AB - The lifetime prevalence of posttraumatic stress disorder (PTSD) is about 8%-14% in the general population, and trauma victimization (51%-98%) and PTSD (up to 42%) are even more prevalent among persons treated within public-sector mental health clinics. Despite this, individuals with PTSD and severe mental illness (SMI) who are treated within the public sector tend to receive inadequate mental health services. In addition, treatments for PTSD for this population remain undeveloped, with virtually no available empirical treatment outcome data to guide clinicians. We propose a model for a comprehensive, multicomponent cognitive-behavioral treatment program for this target population that includes elements of consumer education, anxiety management training, social skills training, exposure therapy, "homework" assignments, and long-term follow-up care. Special considerations for public-sector consumers with PTSD and SMI are addressed, as are directions for future research. PMID- 15334986 TI - Adolescents with attention-deficit/hyperactivity disorder: an overview of empirically based treatments. AB - The author first presents an overview of attention-deficit/hyperactivity disorder (ADHD) as it presents in adolescents. He reviews what is known about the predominantly inattentive subtype in adolescents, the persistence of symptoms into this developmental phase, and comorbid disorders in adolescent patients with ADHD. The author then reviews treatments for adolescents with ADHD for which there is some empirical support in the scientific literature. He first discusses common assumptions concerning the treatment of ADHD and evidence for or against these assumptions. Information on therapies that have been shown to be ineffective or the benefit of which is unproven are then described. These include cognitive-behavioral therapy and social skills training. The author then presents an overview of what is known about the medication treatment of ADHD and discusses how this information is applicable to adolescents with the disorder. Four main classes of drugs are discussed: stimulants, noradrenergic reuptake inhibitors, tricyclic antidepressants, and antihypertensive agents. The author then reviews the use of several psychosocial interventions, including contingency management strategies, parent training in behavior management methods, and teacher training in classroom management, and discusses how these strategies can best be used for adolescents with ADHD. The author then discusses the use of combined treatment with psychosocial interventions and medication. Finally, information on the use of physical exercise as therapy for adolescents with ADHD is discussed. PMID- 15334988 TI - Documentation: the doctor's dilemma. PMID- 15334987 TI - Reboxetine: a norepinephrine selective reuptake pump inhibitor. PMID- 15334989 TI - Killing family members: mental illness, victim risk, and culpability. PMID- 15334990 TI - Family and patient resources. How to help psychiatric patients with cognitive deficits: a guide for families and friends. PMID- 15334992 TI - Macroporous glass-ceramic materials with bioactive properties. AB - In the present research work, glass powders and three different organic starches were used to realize macroporous glass-ceramic scaffolds for bone substitutions. For this purpose, bioactive glass powders belonging to the system SiO2-CaO-Na2O MgO were mixed in a liquid medium with the desired amount of the selected organic phase. Afterwards, by progressively raising the temperature, the water uptake of starches occurred and led to the gelling of the whole system. The resultant gel underwent two thermal treatments in order to eliminate the organic phase and to allow the sintering of the glassy phase. In this way, macroporous glass-ceramic scaffolds were successfully prepared. The samples were characterized by means of optical and scanning electron microscopy with compositional analysis. The volume and mean size of the obtained porosity were investigated by means of mercury intrusion porosimetry, whereas its morphology was assessed by means of microscopic observations. The structure of the original and the resultant materials were investigated by X-ray diffraction. In order to study the reactivity of the scaffolds towards physiological media, the samples were soaked in a simulated body fluid for various times. On their soaked surfaces, scanning electron microscopy and compositional analysis were carried out in order to assess their bioactivity. PMID- 15334991 TI - Stray-field imaging and multinuclear magnetic resonance spectroscopy studies on the setting of a commercial glass-ionomer cement. AB - A commercial glass-ionomer (Fuji II) was studied using different NMR techniques. 1H and 19F stray-field imaging (STRAFI) were used to monitor the curing kinetics of the cement and two processes were distinguished: the gelation and maturation phases. Characterization of the aluminium and silicon species present in the glass component and cement was performed by conventional 27Al and 29Si MAS and two-dimensional 27Al triple quantum MAS NMR (MQMAS) techniques. Quantification of the aluminium in the glass component and in the cement indicates the leaching of about 32% of the 4-coordinate aluminium, about 100% of the 5-coordinate aluminium and about 41% of the 6-coordinate aluminium, during the setting reaction. It is also shown that the 5-coordinate aluminium is only present in the surface layer created by the acid attack during the cement formation process. In the cement, silicon maintains species with four bridging oxygens after the leaching of tetrahedral (4-coordinate) aluminium. The silicon analysis correlates well with the aluminium determinations. PMID- 15334993 TI - Effect of fluoride addition on the properties of dental alginate impression materials. AB - Fluoride-containing dental alginate impression materials can exert a considerable reduction in enamel solubility. The objective was to evaluate the effects of fluoride addition in the alginate impression materials on the properties and subsequent release of fluoride. Four experimental alginate impression materials were studied. Materials were mixed with distilled water (control) or 100-ppm fluoride solution. One or two percent NaF, or 1% SnF2 was added to the materials, which were mixed with distilled water. Fluoride release, flexibility, recovery from deformation, setting time, compressive strength and elastic modulus were determined in accordance with the ISO 1563 and ANSI/ADA Spec. 18. Fluoride release increased after addition of fluoride, and the released amount was 0.762 14.761 ppm. Addition of NaF or SnF2 resulted in higher fluoride release than the control group (p < 0.05). After fluoride addition, flexibility was 15.45-20.27%, and the recovery from deformation did not change except one material. Compressive strength after fluoride addition was 0.36-1.12 MPa. Addition of NaF or SnF2 in an alginate impression material may result in effective release of fluoride without deteriorating the properties of material itself. PMID- 15334994 TI - Wear of surface engineered metal-on-metal hip prostheses. AB - The wear of existing metal-on-metal (MOM) hip prostheses (1 mm3/million cycles) is much lower than the more widely used polyethylene-on-metal bearings (30-100 mm3/million cycles). However, there remain some potential concerns about the toxicity of metal wear particles and elevated metal ion levels, both locally and systemically in the human body. The aim of this study was to investigate the wear, wear debris and ion release of fully coated surface engineered MOM bearings for hip prostheses. Using a physiological anatomical hip joint simulator, five different bearing systems involving three thick (8-12 microm) coatings, TiN, CrN and CrCN, and one thin (2 microm) coating diamond like carbon (DLC) were evaluated and compared to a clinically used MOM cobalt chrome alloy bearing couple. The overall wear rates of the surface engineered prostheses were at least 18-fold lower than the traditional MOM prostheses after 2 million cycles and 36 fold lower after 5 million cycles. Consequently, the volume of wear debris and the ion levels in the lubricants were substantially lower. These parameters were also much lower than in half coated (femoral heads only) systems that have been reported previously. The extremely low volume of wear debris and concentration of metal ions released by these surface engineered systems, especially with CrN and CrCN coatings, have considerable potential for the clinical application of this technology. PMID- 15334995 TI - Characterization of nanostructure phenomena in airborne particulate aggregates and their potential for respiratory health effects. AB - Airborne aggregates of nanoparticulates were collected on carbon/form-coated, 100 mesh Ni TEM grids in a thermal precipitator and observed in an analytical TEM utilizing a BF-SAED-DF-EDS characterization protocol to identify the nanocrystalline or nanoparticulate components, especially their degree of crystallinity, size, structural/morphologic features, and chemistries. Reference aggregates of TiO2 rutile and anatase as well as Si3N4 nanoparticles were used to establish these characterization protocols, which were applied to several hundred individual particulates: homogeneous aggregates of carbonaceous/diesel particulate matter, complex mixtures of carbonaceous matter, including carbon nanocrystals, and inorganic nanocrystals; and heterogeneous, nanocrystal/nanoparticulate aggregates. Most airborne particulates were aggregates ranging in aerodynamic diameters from a few nanometers to a few microns; containing as few as 2 nanocrystals to several thousand nanocrystals or nanoparticulates such as carbonaceous spherules arranged in complex branched homogeneous aggregates composing diesel exhaust, with spherule diameters ranging from 10 to 30 nm. The potential for ultrafine airborne aggregates to fragment into hundreds or thousands of nanoparticulate components in human airways and act as toxic agents in deep lung tissue is demonstrated. PMID- 15334996 TI - Interactions of different types of localized corrosion in surgical implants. AB - Surgical implants often show different types of localized corrosion such as corrosion fatigue cracking, pitting and crevice corrosion on the same part. Interactions of these different corrosion phenomena were investigated. This was done by cyclic loading of electropolished tensile specimens at different constant and changing potentials. Material investigated was a surgical implant steel X2CrNiMo18-15-3 which was immersed in physiological NaCl solution. Pitting and repassivation potentials were determined. Samples with and without artificial cracks as well as masked specimens were tested. Incubation period for first damage, density and size of pits by coulometric and volumetric method were determined. The fracture surfaces were then investigated by SEM. Results show that not in all cases pitting corrosion was the cause for corrosion fatigue cracking. Also pitting is favoured by crack formation. Density of pits increases by a factor of 5 without any change to pitting potential. There are primary pits formed prior to crack initiation and secondary pits formed after crack initiation. At samples without crack there is almost no difference between the optically measured value of total pit volume and the coulometrically determined value. At samples with cracks coulometric volume of pits is much larger than optical one. This proves that there is a significant amount of crevice corrosion in the crack. The corrosion current density in the crack increases by two orders of magnitude when comparing it to electropolished surface of the sample. Results of laboratory experiments are confirmed by failure of a real implant. PMID- 15334997 TI - Cell growth and function on calcium phosphate reinforced chitosan scaffolds. AB - Macroporous chitosan scaffolds reinforced by calcium phosphate powders such as hydroxyapatite (HA) or calcium phosphate invert glass were fabricated using a thermally induced phase separation technique. Human osteoblast-like MG63 cells were cultured on the composite scaffolds for up to 11 days, and the cell growth and function were analyzed. The cell growth is much faster on the chitosan/HA scaffolds incorporated with the glass (CHG) than on the chitosan/HA scaffold without the glass (CH). The total protein content of cells were quantified and increased over time on both composites (CH, CHG) but was significantly higher on CHG after 7 days of culture. The cells on CHG also expressed significantly higher amount of alkaline phosphatase at days 7 and 11 and osteocalcin at day 7 than those on CH. The results suggested that the addition of glass in chitosan/hydroxyapatite composite scaffolds might enhance the proliferation and osteoblastic phenotype expression of MG63 cells. However, the chitosan-matrix scaffolds did not show higher phenotype expression of MG63 cells, in comparison with the TCPS plate, probably due to the degradation of chitosan and release of acidic byproducts. Larger amount of soluble calcium phosphate invert glasses should be added into the scaffolds to prevent chitosan from fast degradation that may affect the differentiation of osteoblast cells. PMID- 15334998 TI - Effects of alkali pretreatment of silk fibroin on microstructure and properties of hydroxyapatite-silk fibroin nanocomposite. AB - Nanocomposites comprising hydroxyapatite (HAp) and silk fibroin (SF) were synthesized from Ca(OH)2 suspension co-dispersed with SF fine particles and H3PO4 solution via a wet-mechanochemical route. The SF particles were modified with an alkali solution to increase contact points between HAp phase and SF matrix. HAp crystallites grow more preferentially along c-axis on alkali pretreated SF substrates. The composites exhibit porous microstructure with 70% of open porosity and about 70% of the interpores ranging from 40 to 115 microm in diameter. The peak shifts in amide II band of SF indicate that the chemical interactions between HAp crystals and SF matrix are intensified by the alkali pretreatment of SF. The stronger inorganic-organic interactions promote the formation of three-dimensional network extending throughout the composites, bringing about an increase of 63% in the Vickers hardness to the composite. PMID- 15334999 TI - Fabrication of 1-dimensional porous hydroxyapatite and evaluation of its osteoconductivity. AB - Porous HA ceramics with 1-dimensional pore channels were fabricated to obtain controllable microstructure. 1-dimensional porous HA was objected to find out the optimum condition of bone ingrowth and also to facilitate the observation of osteocondutive behavior in porous HA. The porous structure was formed by burnt out of polymeric fibers and the size of pores was determined by the diameter of polymeric fibers. The porosity could be varied by the thickness of HA slurry coated on polymeric fiber and by the thickness of HA tapes inserted between fiber layers. As result, 1-dimensional porous HA ceramics of this study have the uniform interconnection size (50-500 microm) and the linearly open pore structure. The compressive strength of 1-dimensional porous HA was 6-10 MPa similar to that of human cancellous bone. On the in vivo test, oteon-like osteoconduction in pore channel of 1-dimensional porous HA was observed, like what had been found in cortical bones. This osteon-like new bone grew from the surface to the center of pore channels. The 1-dimensional porous HA ceramics prepared in this study were very useful as a model system to observe bone ingrowth in the porous HA implants. PMID- 15335000 TI - Fixation of distal femoral osteotomies with self-reinforced poly(L/DL)lactide 70:30/bioactive glass composite rods. An experimental study on rats. AB - Self-reinforced poly(L/DL)lactide 70:30/bioactive glass [SR-P(L/DL)LA/bioactive glass] composite rods, 2 mm in diameter and 36 mm in length, were implanted into the dorsal subcutaneous tissue of 16 rats. Osteotomies of the distal femur were fixed with these rods (2 x 15 mm) in 64 other rats. The follow-up times varied from one week to one year. After sacrifice, three-point bending and shear tests, and molecular weight measurements were performed for subcutaneously placed rods. Radiological, histological, histomorphometrical, microradiographic, and oxytetracycline-fluorescence studies of the osteotomized and intact control femora were performed. At 24 weeks the mechanical properties had decreased significantly. Thirty-nine osteotomies healed uneventfully. One of the 64 evaluated osteotomies showed signs of infection at six weeks, and there were 19 non-unions and six delayed unions. In 20 operations the fixation was loose and out of these 14 non-unions were observed. No gross signs of inflammatory or foreign-body reactions were observed. The present investigation showed that the mechanical strength and fixation properties of SR-P(L/DL)LA/bioactive glass composite rods are suitable for fixation of cancellous bone osteotomies in rats as long as the operative technique is correct. The present article is the first report on the application of SR-P(L/DL)LA/bioactive glass composite rods for fixation of cancellous bone osteotomies. PMID- 15335001 TI - Promoting the cytocompatibility of polyurethane scaffolds via surface photo grafting polymerization of acrylamide. AB - Polyurethane (PU) porous scaffolds were modified by grafting polymerization of acrylamide (AAm) initiated under UV light. A pre-adsorbing-monomer method was used beforehand. FTIR-ATR spectroscopy and X-ray photoelectron spectroscopy (XPS) measurements confirmed the presence of grafted PAAm on PU scaffolds. The measurement of water adsorption demonstrated the improvement of hydrophilicity after PU scaffolds were grafted with polyacrylamide (PAAm). The PAAm grafting degree related to the amount of AAm adsorbed, average pore diameter, and the degree of porosity. This study shows that higher degree of porosity and bigger porous areas yielded larger amounts of absorbed AAm and higher grafting degrees. In vitro human endothelial cell cultures of PU scaffolds modified with hydrophilic PAAm showed better cytocompatibility than the control matrix. PMID- 15335002 TI - Influence of fluorapatite minor additions on behavior of hydroxyapatite ceramics. AB - Fluorinated hydroxyapatite is known to be less soluble by body fluids, resulting in enhanced resistance to biodegradation in vivo conditions, as compared to the pure hydroxyapatite ceramics. The present work was aimed at the investigation of the effect of minor additions of ultrafine fluorapatite (up to 10 wt%) on the sintering behavior and mechanical properties of hydroxyapatite ceramics. In vitro testing for the osteoblast-like cells viability and proliferation was performed with the samples of varying fluorapatite content. It was found that the fluorapatite addition hinders the sintering shrinkage and lowers the strength, but does not generally affect negatively the viability of the cells. PMID- 15335003 TI - Effects of PVA sponge containing chitooligosaccharide in the early stage of wound healing. AB - Poly(vinyl alcohol) (PVA) sponges with different chitooligosaccharide (COS) content were prepared for wound-dressing application. The morphological structure of PVA sponges was observed by scanning electron microscopy. As the concentration of COS-loaded PVA sponge increased, the average pore size of sponge decreased and the release rate of COS from the sponge also slightly decreased. The accelerating effect of the COS-loaded PVA sponges on open wound healing in rats was investigated by macroscopic examination and measurement of wound area. The COS loaded sponges were found to be very effective as a wound-healing accelerator in the early stage of wound healing. The wound treated with the COS-loaded PVA sponge was almost reepithelialized, granulation tissues in the wound were considerably replaced by fibrosis at 8 days after initial wounding. The COS loaded PVA sponge was considered to be a suitable wound-healing formulation due to its easy preparation and high effectiveness. PMID- 15335004 TI - Demand for continuing medical education programs on cancer care among primary care physicians in North Carolina. AB - BACKGROUND: Primary care physicians have a central role in cancer prevention and control services, yet relatively little attention has been given to their needs for continuing medical education (CME) that clarify or update screening guidelines, enhance recognition of signs or symptoms of cancer, and address ongoing health issues in patients treated for cancer (e.g., pain control lymphedema, tertiary prevention). METHODS: A random sample of 600 primary care physicians practicing in urban and rural locations in North Carolina was selected to assess past cancer-related CME sessions, and demand for current cancer education topics. RESULTS: Of 539 eligible, 231 surveys were returned (43%). Approximately 37% of respondents had attended no cancer-related CME in the last two years. Highest interest for cancer CME topics was found for screening for breast and skin cancers, general update diagnostic skills, pain management and patient/family support, side effects from treatment, lymphedema management and lymphedema diagnosis, genetic susceptibility diet and smoking cessation. Interest levels by CME topic did not vary by urban/rural practice settings. CONCLUSION: There is low access but high demand for cancer-related CME topics among primary care physicians. Strategies are needed to fill this need and to assess impact. PMID- 15335005 TI - North Carolina's infant mortality problems persist: time for a paradigm shift. PMID- 15335006 TI - Programs and policies to address the problem of infant mortality: a long history. PMID- 15335007 TI - Improving pre-pregnancy health is key to reducing infant mortality. PMID- 15335008 TI - Infant mortality: 1963 to present medical developments and legislative changes. PMID- 15335009 TI - North Carolina makes strides to reduce SIDS, but challenges lie ahead. PMID- 15335010 TI - Folic acid and birth defects prevention: a public health success story. PMID- 15335011 TI - Addressing perinatal health disparities: another place for a paradigm shift. PMID- 15335012 TI - Making a difference in infant survival: evidence-based actions to reduce tobacco exposure during pregnancy and infancy in North Carolina. AB - North Carolina faces major challenges in dealing with smoking and its consequences during pregnancy and infancy. Evidence-based strategies exist to help pregnant and parenting smokers to quit, to discourage young people from becoming smokers and to reduce exposure of infants to SHS. North Carolina is making progress in implementing these strategies, but more infant lives could be saved each year if the state adopted a more comprehensive approach to addressing tobacco use by improving cessation services for pregnant and parenting smokers, reimbursing clinicians for providing cessation services, increasing state excise taxes on tobacco products, establishing statewide help or quitline services and adopting tobacco-free school policies. These proven strategies can make a difference. PMID- 15335013 TI - Preterm birth in North Carolina. PMID- 15335014 TI - Expanding Medicaid income eligibility for family planning: an opportunity to improve reproductive outcomes and lower Medicaid costs. PMID- 15335015 TI - There is life (and death) beyond the infant year: North Carolina's recent experience in reducing child deaths. PMID- 15335016 TI - Unintended pregnancies in North Carolina. PMID- 15335017 TI - Latino health in North Carolina. PMID- 15335018 TI - Mental health reform. PMID- 15335019 TI - [The use of nutritional antioxidants for the correction of metabolic disorders of cardiovascular patients]. AB - The effectiveness of complex dietary therapy, comprising the use of nutritional antioxidants, for patients with degree I-II essential hypertension and CHD has been evaluated. It is found that the including of soy isoflavones and selenium into the nutrition had a positive effect on the clinical presentation of disease, lipid metabolism, organism antioxidant status, indices of blood coagulation and anticoagulation systems. PMID- 15335020 TI - [Enteral therapeutic nutrition in the complex therapy and prevention of diseases]. AB - Baseline nutritional impairments, patient's malnutrition, and inadequate correction of metabolic disorders considerably reduce the effects of therapeutic measures, increase the risk of septic and infectious complications, negatively influence the hospital length of stay, increase mortality rates. Concomitant and profound disorders of metabolic homeostasis system in various diseases and critical conditions dictate a multi-component approach to the correction of metabolic disorders requiring specially chosen nutrient compositions and the routes of their administration. The development of currently available nutritional formulae was based on the theory of balanced nutrition reflecting the physiological requirements in nutrients and calories of a healthy person, and also taking into account specific features of pathogenesis, clinical course, disease stage, the nature and extent of metabolic disorders, functional condition of the gastrointestinal tract, the effects of specific nutrients on the intensity of metabolic processes. Nutrition adapted to the patient's disease and the digestive tract functional condition provided by diets containing specific nutrients (arginine, glutamine, omega-3 fatty acids) has a number of advantages over standard nutritional formulae, and poses a directed therapeutic effect. The nutritional formulae "Nutrien" are represented by the standard ("Nutrien Standard"), and semi-elemental ("Nutrien Elemental") formulae, as well as by the formulae of directed effect ("Nutrien Hepa", "Nutrien Nephro", "Nutrien Pulmo", "Nutrien Diabet", "Nutrien Immun"). PMID- 15335021 TI - [Correction of selenium deficiency in patients with pneumonia]. AB - The estimation of selenium (Se) safety in patients with pneumonia is important for evaluation of antioxidant defense system capacity and the immune status of the patients. The research has been carried out on the serum Se levels in patients with pneumonia at standard treatment and at treatment with the usage of selenium enriched food supplement (Se-spirulina). The results of research have shown that application of additional selenium in amount providing physiological requirement leads to full varnishing its deficiency cases and to restoration in the majority of patients the suboptimum Se level. PMID- 15335022 TI - [Risk factors and etiological structure of children's cross-allergic reactions to food products]. AB - Authors of article have recently analyzed the frequency of prevalence and risk factors of cross-reactivity to foodstuffs among 239 children of Abakan in the age of from 6 months till 15 years with skin, respiratory and combined manifestation of atopy. It was proved that risk factors of development of cross-reactivity to food among children of the first years of life are hereditary predisposition to allergic diseases (77.8%) and early introductions of supplemental feeding (44%). Among children older then 7 years pollen sensitization (62%) which preceded the development of cross allergic reactions between separate foodstuffs reaches up to 73.7%, between pollen and foodstuff up to 79% and between epidermal and food up to 10.1%. PMID- 15335023 TI - [The hygiene estimation of polyamide shells safety for the sausage products]. AB - The investigation of polyamide shells, contrasting according to the level of penetration and layering, named as "Amitan", "Amiflex T", "Amismok" were carried out. It was stated that in terms of technological process of cooked meats production modeling the studied polyamide shells meet the requirements of hygiene safety. PMID- 15335024 TI - [Vitamin B1 and B2 ratio as a method of brewer's and food yeast identification]. AB - The data on vitamins B1 and B2 content in dry brewer's and baker's yeast are submitted. Their significance as these nutrients source is evaluated. Brewer's yeast are mainly the source of vitamin B1 while baker's yeast--the source of vitamin B2. 5 g of yeast give 10 per cent of vitamin B1 and B2 daily recommended allowance correspondingly. Vitamin B1 and B2 ratio has been proposed as a identification criteria of brewer's and baker's yeast. Ratio more than 1.0 gives evidence that this is brewer's yeast. Vitamin B1 content 4 mg per 100 g or more and at the same time vitamin B1 and B2 ratio exceeding 3.0 demonstrate high quality of brewer's yeast. PMID- 15335025 TI - [The peculiaries of the nutrition and immune status of people in the condition of penitentiary establishment]. AB - The decrease of calorie in ration content, insufficient consumption of proteins and lipids was found among the patients in the condition of penitentiary establishment, it results in decrease of characteristics of natural immunity and resistibility to various infections agents, to the action of toxic substances, stress and other negative conditions that take place in penitentiary establishment a disbalanse of regulatory subpopulation of cellular consistent of immunity and predominance of suppressive-cytotoxic influence was observed among the patients of the penitentiary establishment (with a reduced ration of nutrition) over halt a year. Considering a health problem, in the establishments of imprisonment one may probably state that a reduced ration of nutrition is one of the main courses of immune homeostasis shift leasing in the basis of specific and non-specific phenomena of morbidity. PMID- 15335026 TI - [Biologically active food supplements. Report 1]. AB - In article we would like to remind once of what each of the food supplement consumer should know. Food supplement is not a medicine, you can treat no illness with it. But at the same time we should understand, that it is impossible to live without them, as well as it impossible to live without food or the air. It is important to deepen one's knowledge of food supplements: what they consist of, which of them are health-giving and which are not. PMID- 15335027 TI - [The inhibition of the food lipid radical oxidation by the flavonoid antioxidants]. AB - There are the current events on the oxidative damage of the food lipids summarized. The theoretical grounds of the inhibition of free radical oxidation by phenol compounds is proposed. It includes the electronic mechanism antioxidants of the antioxidative action of natural flavonoid substances. PMID- 15335028 TI - [On the mechanisms of the alcohol action on human organism]. AB - In article is discussed the information on properties of alcohol, distribution in organism, systems of oxidation, elimination, biological activity of ethanol and acetaldehyde, mechanisms of damaging action of an alcoholic intoxication. PMID- 15335029 TI - [The history of the development of the children nutritiology at the Institute of Nutrition of RAMS]. AB - The history of infant and child nutrition department of the Institute of Nutrition of RAMS is briefly reviewed. The most important results of scientific investigations performed at the department from 1930 year till present time as well as methodical materials prepared by department specialists and serving as basis of official Ministry of Health recommendations and rules of infant and child nutrition, are discussed. Special attention is drawn to the problem of infant nutrition. PMID- 15335030 TI - Unifying separation science: EuSSS. PMID- 15335031 TI - Dual column capillary gas chromatographic system for the in situ analysis of volatile organic compounds on a cometary nucleus. AB - Two Wall Coated Open Tubular capillary columns, coated with poly(cyanopropylphenyl-dimethyl)siloxane and poly(diphenyl-dimethyl)siloxane stationary phases, have been selected for use in the COmetary SAmpling and Composition space experiment for the separation and identification of the wide range of volatile organic compounds which could be present in cometary nuclei. This article presents the main characteristics of the tandem column system for the analysis of solutes of cometary interest within the constraints of space instrumental operating conditions. The high efficiency of the columns is demonstrated and the influence of the operating conditions on their separation properties are investigated. The studied columns exhibit complementary retention pattern: their use in a dual column system makes it possible to achieve the separation and the identification of the compounds of interest. Finally, the good analytical behavior of the columns when analyzing samples which include large amounts of water, the main presumed volatile in comets, is demonstrated. The presented results thus show the suitability of the selected tandem columns system for the desired analyses, and their performance on adaptation to in-situ cometary chemical investigation. PMID- 15335032 TI - Effects of pressure drop on absolute retention matching in comprehensive two dimensional gas chromatography. AB - Comprehensive two-dimensional gas chromatography (GC x GC) analysis has the capability to resolve many more components of complex mixtures than traditional single column GC analysis. There is an increasing need to provide reliable identification of these separated components; time-of-flight mass spectrometry (TOFMS) is the most appropriate technology to achieve this task. Rather than require MS for all GC x GC separations, it is desirable to assign peak identities to specific peak positions in the GC x GC separation space, and this necessitates matching peak retentions in the two experiments - GC x GC-FID and GC x GC-TOFMS. The atmospheric vs. vacuum outlet conditions confound this task. It is shown here that by employing a supplementary gas supply, provided to a T-union between the column outlet and the MS interface, it is possible to generate 2D chromatograms for GC x GC-FID and GC x GC-TOFMS that are essentially exactly matched. There is no degradation in separation performance or efficiency in the second column in the system interfaced to the T-union. Since the GC x GC-FID experiment uses hydrogen for maximum efficiency, and GC x GC-TOFMS uses helium carrier, translation of (conditions/retentions) must account for the different viscosities of the carrier gases. Translation of conditions is based on well-known principles established in single column analysis. Tabulated data illustrate that retention reproducibility was of the order of better than 4 s for the average first dimension retention difference, and about 40 ms for the average second dimension retention difference when comparing GC x GC-FID and GC x GC-TOFMS results. This should provide considerable support for identification in routine GC x GC-FID analysis of specific sample types, once the peaks in 2D separation space have been assigned identities through GC x GC-TOFMS analysis. PMID- 15335033 TI - Composition of the black crusts from the Saint Denis Basilica, France, as revealed by gas chromatography-mass spectrometry. AB - The organic fraction of black crusts from Saint Denis Basilica, France, is composed of a complex mixture of aliphatic and aromatic compounds. These compounds were studied by two different analytical approaches: tetramethyl ammonium hydroxide (TMAH) thermochemolysis in combination with gas chromatography mass spectrometry (GC-MS), and solvent extraction, fractionation by silica column, and identification of the fraction components by GC-MS. The first approach, feasible at the microscale level, is able to supply fairly general information on a wide range of compounds. Using the second approach, we were able to separate the complex mixture of compounds into four fractions, enabling a better identification of the extractable compounds. These compounds belong to different classes: aliphatic hydrocarbons (nalkanes, n-alkenes), aliphatic and aromatic carboxylic acids (n-fatty acids, alpha,omega-dicarboxylic acids, and benzenecarboxylic acids), polycyclic aromatic hydrocarbons (PAH), and molecular biomarkers (isoprenoid hydrocarbons, diterpenoids, and triterpenoids). With each approach, similar classes of compounds were identified, although TMAH thermochemolysis failed to identify compounds present at low concentrations in black crusts. The two proposed methodological approaches are complementary, particularly in the study of polar fractions. PMID- 15335034 TI - Structural elucidation of catechin and epicatechin in sorrel leaf extracts using liquid-chromatography coupled to diode array-, fluorescence-, and mass spectrometric detection. AB - Flavonoids are natural compounds in medicinal herbs and posses several biological activities important in plant drug design. Especially strong antioxidant effects play an important role against radical oxidative stress causing pathological processes, such as arteriosclerosis or cancer. The aim of this work was to investigate unknown analytes found in sorrel leaf (Rumex acetosa) extracts in order to discover new leading compounds to enable quality control in phytopharmaceuticals made thereof. Therefore compounds of interest were separated after methanolic extraction by reversed-phase liquid chromatography (RP-LC) employing silica-C18 as stationary phase using gradient elution with water (10 mM H3PO4) and acetonitrile as mobile phase. Structural elucidation was carried out by diode array, fluorescence, and mass spectroscopic detection. Photodiode-array detection allowed the extraction of UV-absorbance spectra from the peaks of interest. Absorbance maxima were found at 203 and 279 nm with a shoulder at 230 nm. Additionally fluorescence emission and excitation spectra were recorded from the analytes using a fluorescence detector (FLD) after stop flow (lambdaex = 279 nm, lambdaem = 307 nm). The chromatographic reversed-phase system was coupled to an ion-trap mass spectrometer (LC-MS) by using an electrospray ionization interface (ESI). After optimization processes the separation was carried out using an ammonium acetate buffer at pH 5.5. Mass spectrometric detection turned out to be more sensitive in negative mode. Collisionally induced dissociation (CID) was used to obtain fragment ions of structural relevance (LC-MS/MS). Finally, compounds of interest coming from sorrel leaf (Rumex acetosa) eluting at low acetonitrile concentrations were confirmed to be catechin and epicatechin. PMID- 15335035 TI - Sequential injection chromatographic determination of paracetamol, caffeine, and acetylsalicylic acid in pharmaceutical tablets. AB - In this contribution, a new separation method for simultaneous determination of paracetamol, caffeine, acetylsalicylic acid, and internal standard benzoic acid was developed based on a novel reversed-phase sequential injection chromatography (SIC) technique with UV detection. A Chromolith Flash RP-18e, 25-4.6mm column (Merck, Germany) and a FIAlab 3000 system (USA) with an 8-port selection valve and a 5 mL syringe were used for sequential injection chromatographic separations in our study. The mobile phase used was acetonitrile-(0.01 M) phosphate buffer (10:90, v/v) pH 4.05, flow rate 0.6 mL min(-1). UV detection was at 210 and 230 nm. The validation parameters showed good results: linearity (r >0.999) for all compounds, detection limits in the range 0.3-0.8 microg mL(-1), repeatability (RSD) of peak heights between runs in the range 1.10-4.30% at three concentration levels and intra-day repeatability of the retention times in the range 0.28 0.43%. The analysis time was <6 min. The method was found to be applicable for the routine analysis of the active compounds paracetamol, caffeine, and acetylsalicylic acid in pharmaceutical tablets. PMID- 15335036 TI - A liquid chromatography-mass spectrometric method for the determination of oak moss allergens atranol and chloroatranol in perfumes. AB - This paper describes a validated liquid chromatographic-tandem mass spectrometric method for quantitative analysis of the potential oak moss allergens atranol and chloroatranol in perfumes and similar products. The method employs LC-MS-MS with electrospray ionization (ESI) in negative mode. The compounds are analysed by selective reaction monitoring (SRM) of 2 or 3 ions for each compound in order to obtain high selectivity and sensitivity. The method has been validated for the following parameters: linearity; repeatability; recovery; limit of detection; and limit of quantification. The limits of detection, 5.0 ng/mL and 2.4 ng/mL, respectively, for atranol and chloroatranol, achieved by this method allowed identification of these compounds at concentrations below those causing allergic skin reactions in oak-moss-sensitive patients. The recovery of chloratranol from spiked perfumes was 96+/-4%. Low recoveries (49+/-5%) were observed for atranol in spiked perfumes, indicating ion suppression caused by matrix components. The method has been applied to the analysis of 10 randomly selected perfumes and similar products. PMID- 15335037 TI - Separation, identification, and quantification of selected withanolides in plant extracts of Withania somnifera by HPLC-UV(DAD)--positive ion electrospray ionisation-mass spectrometry. AB - This paper describes a method for separation, identification, and quantification of selected withanolides in Withania somnifera plant extracts by HPLC-UV(DAD) Mass Spectrometry (HPLC-MS). Withaferin-A (WS-3), 12-deoxywithastramonolide (WS 12DS), Withanolide A (WS-1), and Withanone (WS-2) were used as external standards. The compounds were isolated from Withania somnifera by repeated column chromatography of the root extract and their identity was established by 1H- and 13C-NMR and mass spectral data. The compounds were chromatographed on a Merck (250 x 4.6 mm ID, 5 microm) column and analyzed by Electrospray Ionization on a mass spectrometer in Selected Ion Mode (SIM). For quantification, [M + Na]+ ions were monitored. Linear calibration curves were obtained in the concentration range of 1.50 microg/mL to 6.5 microg/mL. The method was applied successfully to the detection and quantification of the said withanolides in a number of samples. PMID- 15335038 TI - Screening of oxazepine indole enantiomers by means of high performance liquid chromatography with imprinted polymer stationary phase. AB - Chromatographic enantiomer separations of different oxazepine indole derivatives were performed using a molecularly imprinted polymer. A 5aR,12R,13S-trans-6,6 dimethyl-12,13-dihydro-6H-5a, 1 3-methanoindolo[2,1-b][1,3]naphthoxazepine-12 carboxamide enantiomerderivative was used as a template and the resultant polymer has shown enantiomer recognition for series of template related compounds. The mechanistic description of the chiral discrimination process is scrutinised, comparing the discrimination between the different conformations and substituents of the oxazepine indoles. PMID- 15335039 TI - Sample preparation followed by HPLC under harmless 100% aqueous conditions for determination of oxytetracycline in milk and eggs. AB - A simple and hazardous chemical-free method for the high-performance liquid chromatographic determination of oxytetracycline (OTC) residues in milk and eggs has been developed. Sample preparation consists in homogenization with an aqueous solution by means of a handheld ultrasonic homogenizer followed by centrifugal ultrafiltration. HPLC is performed with an isocratic aqueous mobile phase and a photodiode array detector. Average recoveries of OTC (0.05, 0.1, and 0.2 microg mL(-1) for milk; 0.1, 0.2, and 0.4 microg mL(-1) for eggs) were > or =84% with relative standard deviations of < or =2.3%. The total time required for the analysis of one sample and LOQs were <30 min and <0.1 microg mL(-1), respectively. In all the processes, no organic solvents or hazardous reagents were used. PMID- 15335040 TI - Definite peak identification of (R)-and (S)-methadone and (R)- and (S)-EDDP using established HPLC and CE methods. AB - Methadone is widely used for the treatment of opioid dependence. HPLC and CE are widespread methods for drug monitoring and metabolism studies. Although the methods are widely used for methadone and its main metabolite EDDP [1, 2], a definite direct peak identification for EDDP enantiomers is not described. This study describes a method for specific identification of each peak in the chromatogram and electropherogram of methadone analysis. The result of the study demonstrates differences in the elution order of the enantiomers of methadone and EDDP due to the technique used for analysis. The elution order of EDDP using HPLC is interchanged with respect to the order of the peaks in the electropherogram. PMID- 15335041 TI - Determination of the purity of ampicillin by micellar electrokinetic chromatography and reversed phase liquid chromatography on a monolithic silica column. AB - A micellar electrokinetic chromatographic (MEKC) method and a fast reversed-phase liquid chromatographic one have been developed for determining the purity of ampicillin. MEKC separation of ampicillin and its related substances was performed with the use of an untreated fused-silica capillary and 40 mM phosphate borate buffer, pH 7.5 containing 75 mM SDS. The HPLC method employed a monolithic silica C18 column and a mobile phase composed of phosphate buffer, pH 5.2 and ACN, the flow rate being 4.0 mL/min. Both methods were successfully validated. Linearity, relative response factors, limits of quantitation, intermediate precision, and accuracy were evaluated. The methods proved to be fast, reliable, and sufficiently sensitive and, accordingly, well-suited for control of purity of ampicillin substance, injections, and capsules. A combination of both methods can be very useful in the confirmation of impurity profiles. PMID- 15335042 TI - Separation and determination of active components in Radix Salviae miltiorrhizae and its medicinal preparations by nonaqueous capillary electrophoresis. AB - A nonaqueous capillary electrophoresis (NACE) method was developed for simultaneous assay of three bioactive components (1: cryptotanshinone; 2: tanshinone IIA, and 3: tanshinone I) in Radix Salviae miltiorrhizae and in its herbal preparations for the first time. After optimization of separation conditions, a buffer of 250 mmol L(-1) ammonium acetate containing 30% acetonitrile and 1.0% acetic acid (V:V) in methanol was selected for separating the three analytes, but baseline separation of tanshinon I and tanshinone IIA was not obtained. Therefore second-order derivative electropherograms were applied for resolving overlapping peaks. Regression equations revealed good linear relationships (correlation coefficients 0.9943-0.9991) between peak heights in second-order derivative electropherograms and concentrations of the three analytes. The relative standard deviations (RSD) of the migration times and the peak height of the three constituents were in the range of 0.81 -0.88% and 0.34 1.13% (intra-day), 1.57-1.86% and 3.05-5.52% (inter-day), respectively. The recoveries of three constituents ranged from 90.2 to 108.5%. The results indicated that baseline separation of the analytes was sometimes hard to obtain and second-order derivative electropherograms were applicable for the resolving and analysis of overlapping peaks. PMID- 15335043 TI - Sensitivity enhancement of chlorinated phenols by continuous flow liquid membrane extraction followed by capillary electrophoresis. AB - This paper describes a new analytical system, based on the combination of continuous flow liquid membrane extraction (CFLME) enrichment and capillary electrophoresis (CE) separation, for analysis of chlorinated phenols in water samples. Five chlorinated phenols including 3-chlorophenol (3CP), 4-chlorophenol (4CP) 2,4-dichlorophenol (DCP), 2,4,6-trichlorophenol (TCP), and pentachlorophenol (PCP) were separated by CE with Tris/sodium dihydrogen phosphate solution containing methanol 1% (v/v) as the run buffer. CFLME related parameters were investigated and optimal enrichment was obtained by using 0.3 mol L(-1) Tris as acceptor and with a sample pH 5.0, a sample flow rate of 4.0 mL min(-1), and an enrichment sample volume of 150 mL. The detection limit (S/N= 3) was 6.9, 1.0, and 1.7 ng mL(-1) for DCP, PCP, and TCP, respectively. The reproducibility (RSD%, n = 6) was 5.7 for DCP, 2.5 for PCP, and 2.8% for TCP (n = 6). The proposed method was applied to the determination of chlorinated phenols in spiked water samples with relatively satisfactory recoveries. PMID- 15335044 TI - Spectral correlative chromatography and its application to analysis of chromatographic fingerprints of herbal medicines. AB - A signal-processing method known as spectral correlative chromatography (SCC) for two-dimensional data obtained from hyphenated chromatography is developed and applied to chemical chromatographic fingerprint data sets of herbal medicine under specific experimental conditions. The method can judge the presence or absence of a spectral correlative peak among the spectrochromatograms. A local least squares regression model (LLS) is constructed in a piecewise manner to correct the shifts of retention time of some peaks of interest in the chromatograms of various test samples. The results compare favorably with those obtained by a two-point calibrated algorithm. It is shown that performing SCC and LLS on the piecewise clusters of various chromatographic fingerprints is more helpful in practice in revealing their common nature and for characterizing the chemical constituents. This approach holds great potential for facilitating quality control of herbal medicines. PMID- 15335045 TI - Chemical extraction and direct adsorptive purification of recombinant human antigen Ro52. AB - The antigenic protein Ro52 was expressed in the E. coli system harboring a 6 x His tag in the form of insoluble inclusion bodies. Direct chemical extraction of the product using 6-8 M urea proved to be effective. Furthermore, the tagged protein was recovered by direct adsorption on Ni2+-loaded commercial adsorbents derivatized with iminodiacetic acid. Screening experiments in small packed columns revealed that selective binding and elution were possible using a denaturing buffer at pH 4.5. The hydrodynamic evaluation of scaled-up fluidized systems showed values for the phi (dynamic zone) parameter in the range 0.95-1.00 for fluidization in buffer and in the range 0.70-0.85 for the biomass-containing feedstock. Removal of macromolecular DNA released by the disrupted biomass was mandatory. Under optimized process conditions good recovery (60-70%) was achieved and a highly purified (95%) product obtained. The purified Ro52 retained its immunoreactivity against sera of patients with systemic lupus erythematosus (SLE) and Sjogren's syndrome-related disorders. The production and application of new recombinant antigens may contribute to increasing the sensitivity and specificity of the detection of anti-Ro antibodies in these autoimmune diseases. PMID- 15335046 TI - Miniaturised sample preparation of fatty foodstuffs for the determination of polychlorinated biphenyls. AB - A miniaturised analytical method allowing the exhaustive extraction of environmentally relevant polychlorinated biphenyls (PCBs) from fatty foodstuffs and the purification of the extracts in a single step has been developed. After dispersion of the freeze dried sample on silica modified with 44% (w/w) of sulphuric acid, the mixture was packed in a glass column on top of a multilayer silica column used for removal of the lipids and biogenic co-extracted material. Using this arrangement, a complete sample preparation can be accomplished by two successive 10 min static extractions with hexane followed by a brief dynamic step to ensure purging of the sample and sorbents. The analytical method showed a satisfactory performance, with recoveries of the endogenous PCBs studied in the 81-134% range of those found using a more conventional off-line procedure, even though as small an amount of sample as 0.1 g was used. Detection limits by gas chromatography with micro-electron capture detection (GC-ECD) were in all cases lower than 0.3 ng/g sample (freeze dried basis) and the repeatability of the complete analytical procedure better than 14% (except for PCB 167). When combined with GC and ion trap detection in the tandem mass spectrometry mode, the miniaturised method has been proved to be a valuable alternative to the more expensive high resolution mass spectrometry for fast screening of PCBs 77, 126, and 169, even if these congeners were not isolated from the bulk of PCBs. PMID- 15335047 TI - Preconcentration of the herbicide glyphosate and its metabolite AMPA by immobilised metal ion affinity chromatography (IMAC). AB - A method based on Immobilised Metal Ion Affinity Chromatography (IMAC) using a chelating resin (Spheron Oxine 1000) loaded with Pd(II) is evaluated for the extraction and preconcentration of glyphosate and AMPA from natural water samples. The efficiency of the metal-loaded resin in retaining glyphosate and AMPA and the ability of different reagents to recover them is investigated. The most effective elution solution for the recovery of both analytes from the resin is found to be a mixture of 0.1 M HCI and 1 M NaCl. The effect of flow rate, analyte concentration, and sample volume is evaluated. The optimised experimental conditions are then used in the extraction of the analytes from spiked natural waters. The use of the Pd loaded resin led to recoveries ranging from 80-90% for glyphosate and 60-80% for AMPA. PMID- 15335048 TI - Characterization of alkyl-cobalamins formed on trapping of epoxide metabolites of 1,3-butadiene. AB - Analytical methods facilitating studies of electrophilically reactive and genotoxic compounds in vitro and in vivo are needed. The strong nucleophile, cob(I)alamin, formed by reduction of Vitamin B12 [cob(III)alamin], may be used for trapping and analysis of 1,2-epoxides and other electrophiles. In the present study, cob(I)alamin is evaluated as an analytical tool for 1,2-epoxide metabolites (oxiranes) of 1,3-butadiene. Products of reaction of cob(I)alamin with 1,2-epoxy-3-butene (EB), 1,2:3,4-diepoxybutane (DEB), and 1,2-epoxy-3,4 butanediol (EBdiol) have been analyzed by reversed phase high performance liquid chromatography (HPLC) coupled on-line to electrospray ionization mass spectrometry (ESI-MS) and ultraviolet diode array detection (UV-DAD). It was shown that a specific alkyl-CbI complex is formed for each metabolite and that it was possible to discriminate between the products by HPLC-UV and by LC-MS. Quantification of DEB with the method by use of another 1,2-epoxide as an internal standard was successfully performed. The possibility of using cob(I)alamin for trapping and analysis of the three oxirane metabolites of 1,3 butadiene will facilitate quantitative comparisons of species in vitro with regard to metabolism of 1,3-butadiene. PMID- 15335049 TI - Evaluation of a new solid-phase cartridge for the preconcentration of phenolic compounds in water. AB - The aim of this study is to evaluate the efficiency of a new solid-phase extraction cartridge, Spe-ed Advanta, in the extraction and preconcentration of four phenolic compounds (phenol, 2-chlorophenol, 2-nitrophenol, and 2,4 dichlorophenol) from water. The solid phase is a polystyrene-divinylbenzene resin modified with carboxylic groups, these polar groups improve the contact between the matrix and the aqueous solutions in the extraction of polar analytes. We studied several elution solvents in order to find the most efficient one. Sample concentration, sample volume, and sample pH are also investigated as well as the best method for drying the cartridge. Recoveries achieved with the new phase are compared with those obtained with Isolute ENV+, a non-modified polystyrene divinylbenzene stationary bed. The best experimental conditions were then used for determination of the phenols in spiked environmental waters. PMID- 15335050 TI - Quantitative analysis of caffeic acid phenethyl ester in crude propolis by liquid chromatography-electrospray ionization mass spectrometry. AB - Caffeic acid phenethyl ester (CAPE), an active component of propolis from honeybee hives, is known to have antimitogenic, anticarcinogenic, antinflammatory, and immunomodulatory properties. The paper describes a rapid and simple liquid chromatography-electrospray ionisation mass spectrometry method for qualitative and quantitative determination of CAPE. The chromatographic separation was performed with a Luna RP-C18 column using a water-acetonitrile linear gradient. The method was linear over a 0.125-80 ng/mL range (LOD = 62.5 pg/mL). The method was applied for the quantitation of caffeic acid phenethyl ester in crude propolis samples, which were analysed directly after extraction with ethyl acetate solution. PMID- 15335051 TI - Parallel separations of oligonucleotides with optically gated sample introduction on multichannel microchips. AB - With the release of the human genome sequence, there has been increasing attention given to other genetic analyses, including the detection of genetic variations and fast sequencing of multiple samples for pharmacogenomics studies. Rapid injections of samples in multiplexed separation channels by optically gated sample introduction are shown here for DNA separation. Serial separations of four amino acids are shown in less than four seconds on a microchip with four multiplexed channels. Five short oligonucleotides have also been rapidly separated in 2% LPA with four channels using this technique. In addition, multiple unique samples have been simultaneously separated and five-base resolution has been demonstrated. PMID- 15335052 TI - Improving resolution for channel-format chip-based electrophoresis with electrochemical array detection. AB - Resolution in channel electrophoresis has been improved by means of the addition of a surfactant to the running buffer and minimization of the channel internal height and sampling capillary internal diameter. Micellar electrokinetic channel chromatography with electrochemical detection has been applied to the separation of several cationic catecholamines and has been used to continuously monitor a dynamic system of dopamine, norepinephrine, and epinephrine. Resolution was also enhanced by coupling small internal-diameter (5 microm) sampling capillaries with sub-micrometer internal-height separation channels. The improvements in resolution offered by these methods will extend the applicability of channel electrophoresis with electrochemical detection to more complex samples while permitting sample volumes in the nL range to be probed. PMID- 15335053 TI - Differential effects of organic modifiers on the enantioseparation of dimetindene maleate with carboxymethyl-beta-cyclodextrin in capillary electrophoresis. AB - Methanol enhances the enantioresolution of dimetindene enantiomers with carboxymethyl-beta-cyclodextrin (CMCD) as chiral selector at a concentration below its optimal value. The same effect was observed with ethanol (EtOH), although less pronounced. On the other hand, the addition of isopropanol (IP) or acetonitrile (ACN) decreases the enantioseparation. To elucidate the underlying mechanisms of these observed effects, other neutral (beta-CD, hydroxypropyl-beta CD, and trimethyl-beta-CD) as well as chargeable (carboxyethyl-beta-CD and succinyl-beta-CD) CD derivatives were also tested with MeOH as organic modifier. It can be concluded that the increased enantioresolution of dimetindene enantiomers was only noted with CMCD as chiral selector and a short-chain organic modifier containing an alcohol function. The slight deprotonation of CMCD at pH 3.0 was only partly responsible for the high enantioselectivity and the 'favourable' effect of MeOH or EtOH. An important feature that can be concluded from these results is that for this particular analyte approximately the same resolution can be obtained with a lower CMCD concentration and the addition of some MeOH, compared to a MeOH free buffer. PMID- 15335054 TI - Analysis of low molecular weight aldehydes in air samples by capillary electrophoresis after derivatization with 4-hydrazinobenzoic acid. AB - This work reports the analysis of selected aldehydes in air samples using capillary electrophoresis (CE). The method is based on the reaction of aldehydes with 4-hydrazinobenzoic acid (HBA) to give the corresponding hydrazones with maximum absorbance at 290 nm. Under optimized CE conditions, the HBA derivatives of four carbonyls (formaldehyde, acetaldehyde, propionaldehyde, and acrolein) were completely separated from one another, in less than 6 min, using a pH 9.3 tetraborate buffer at 0.040 mol L(-1) concentration as background electrolyte. A few method validation parameters were determined revealing good migration time repeatability (< 1.5% CV) and area repeatability (< 2% CV), excellent linearity (50-300 microg/L, r > 0.996) and adequate sensitivity for environmental applications. The limits of detection with respect to each single aldehyde were in the range of 2.7-8.8 ng L(-1). The methodology was applied to the determination of aldehydes indoors. Samples were collected in HBA impregnated octadecylsilica cartridges, at different times during the day. The most abundant carbonyls in the samples were acetaldehyde followed by formaldehyde, with estimated peak concentrations of 4.3 and 2.9 ppbv, respectively. PMID- 15335055 TI - Development of a capillary zone electrophoretic method to determine six antidepressants in their pharmaceutical preparations. Experimental design for evaluating the ruggedness of method. AB - A capillary zone electrophoresis method is proposed for the separation of six of the antidepressants most used for the treatment of mental illness (clomipramine, paroxetine, fluoxetine, citalopram, fluvoxamine, and trazodone). Optimum conditions for their separation were investigated. A background electrolyte solution consisting of 50 mM phosphate buffer adjusted to pH 2.0, hydrodynamic injection, and 25 kV as separation voltage were used. Relative standard deviations (RSD) were <0.38% and <2.93% for migration time and corrected peak area (n = 24), respectively. Detection limits obtained for the six antidepressants ranged from 0.03 to 0.11 mg L(-1). Stability of the solutions, linear concentration range, accuracy, and precision were examined during validation of the method. A ruggedness test of this method was performed using the fractional factorial model of Plackett-Burman, requiring in our case design of a matrix of 15 experiments, in which the influence of seven factors at three different levels was tested on different electrophoretic results: efficiency; resolution; and corrected peak area. Statistical evaluation of electrophoretic results was achieved by Youden and Steiner's method. The described method is rapid, sensitive, and rugged and it was tested for the pharmaceutical formulation analysis, giving recoveries between 95.6 and 99.1% with respect to the nominal content. PMID- 15335056 TI - Retention behaviour of unsaturated fatty acid methyl esters on porous graphitic carbon. AB - The eluotropic strength of binary mobile phases was calculated for three homologous series of cis, trans, and cis-cis unsaturated fatty acid methyl esters (FAMEs). Binary mobile phases with chloroform, dichloromethane, or tetrahydrofuran as strong solvent and methanol or acetonitrile as weak solvent were tested. The volume fraction of strong solvent in the binary phases was between 0.3 and 0.8. Curves of eluotropic strength versus volume fraction of strong solvents showed similar trends to previously published results for saturated homologues. Correlation coefficients of the plots of eluotropic strength values for saturated versus unsaturated FAMEs were close to 1.0. Therefore these similarities validate the model of eluotropic strength previously established with saturated FAMEs as relevant for unsaturated FAMEs. The separation factors between cis and trans homologues always showed elution of the cis before the trans homologue. The difference in retention is due primarily to the geometry of the molecule. The retention is lowered more by the addition of a first carbon double bond than by the addition of a second one, independently of the mobile phase composition. PMID- 15335057 TI - Using analytical multidimensional isocratic HPLC methods of separation to isolate active constituents in natural products. AB - Multidimensional reversed-phase HPLC was employed to isolate the active constituents from a crude extract of Clerodendrum floribundum R. Br. These active constituents were known to have bioactivity against the enzyme xanthine oxidase and potentially could be employed in the treatment of gout. Using a multidimensional separation approach, rapid isolation of the active constituents was achieved from a complex sample matrix. As each separation dimension was isocratic, no equilibration time was required and consequently the technique shows promise in the scale up to preparative levels where high throughput is important. PMID- 15335058 TI - Development of a solid phase extraction method for simultaneous determination of corticoids and tranquilizers in serum samples. AB - A simple and fast solid phase extraction (SPE) method allowing the preconcentration, clean-up, and subsequent separate elution of phenothiazines (chlorpromazine hydrochloride, acetopromazine, and propionylpromazine hydrochloride) and glucocorticoids (dexamethasone, betamethasone acetate, and phenylbutazone) from serum samples has been developed. Both fractions were separately collected and analyzed without any additional treatment by high performance liquid chromatography with UV-Vis. The performance of the complete procedure was satisfactory irrespective of the spiking level with recoveries in the range 64-85% for all analytes investigated but for phenylbutazone (20%). Repeatability, evaluated as the relative standard deviation, was globally better than 12%. LC-MS was used for final confirmation of the results. PMID- 15335059 TI - Pressurized hot water extraction of insecticides from process dust--comparison with supercritical fluid extraction. AB - Pressurized hot liquid water and steam were used to investigate the possibilities of extracting insecticides (carbofuran, carbosulfan, and imidacloprid) from contaminated process dust remaining from seed-pellet production. Extraction temperature was the most important parameter in influencing the extraction efficiency and rate of extraction, while varying the pressure had no profound effect. A clean-up procedure of the water extracts using solid phase extraction (SPE) was found to be necessary prior to final analysis by high-performance liquid chromatography (HPLC). Quantitative extraction (compared to a validated organic solvent extraction method) of imidacloprid was obtained at temperatures of 100-150 degrees C within 30 min extraction time. Temperatures above 150 degrees C were required to extract carbofuran efficiently. The most non-polar analyte of the investigated compounds, carbosulfan, gave no detectable concentrations with pressurized hot water extraction (PHWE). One reason might be its low solubility in water, and when attempts are made to increase its solubility by increasing the temperature it may degrade to carbofuran. This can explain recovery values above 100% for carbofuran at higher temperatures. A comparison of the PHWE results and those obtained with supercritical fluid extraction (SFE) revealed that PHWE is advantageous for polar compounds, where the solubility of the analyte in water is high enough that lower temperatures can be used. For non-polar compounds carbon dioxide based extraction is preferred unless the target analyte is highly thermostable. PMID- 15335060 TI - Local round robin test for determination of aromatics in diesel fuels by HPLC. AB - Determination of aromatics in diesel fuels by HPLC is covered by European Standard EN 12916 (April 2000). The Standard is based on former standards IP 391/90 and IP 391/95. The method used for measurements is NP HPLC and separation is performed on silica or modified silica columns with RI detection. The precision of the method was checked by determining the repeatability and reproducibility. The content of aromatics was measured in a number of commercial samples. The measurements have been performed in three different laboratories and under different working and instrumental conditions. Satisfactory agreement was established for repeatability and was slightly poorer for reproducibility. The content of polyaromatics is within the tolerance range (IP 590/99) and the content of tricyclic and higher aromatics (tri+) is close to the limit of quantitition in some samples. Clear differences in the content of polycyclic aromatics in domestic diesel and eurodiesel fuels were observed. The results have confirmed the ruggedness and robustness of the measuring method. PMID- 15335061 TI - Simultaneous determination of tretinoin and clindamycin phosphate and their degradation products in topical formulations by reverse phase HPLC. AB - A new HPLC method based on reverse phase separation and photodiode-array detection has been developed for the simultaneous determination of tretinoin and clindamycin phosphate, and their degradation products in topical formulations. The method has been shown to be stability indicating, accurate, and precise for two different formulation vehicles. Separation was achieved on a reverse phase C18 column (Lichrospher, RP18, 5 microm, 25 cm x 4.6 mm ID, Phenomenex, USA) using a simple gradient with aqueous-acetonitrile and aqueous-methanol mobile phases. The method recovery averaged 100.3% for tretinoin and 99.6% for clindamycin phosphate at a concentration range between 80% and 120% of the label claim. The method can be applied to assess the stability of tretinoin and clindamycin phosphate in pharmaceutical formulations containing tretinoin and clindamycin phosphate individually or in combination as active drugs. PMID- 15335062 TI - Approaches to identifying and quantifying polycyclic aromatic hydrocarbons of molecular weight 302 in diesel particulates. AB - Among the PAH class of compounds, high molecular weight PAH are now considered as relevant cancer inducers, but not all of them have the same biological activity. However, their analysis is difficult, mainly due to the presence of numerous isomers and due to their low volatility. Retention indices (Ri) for 13 dibenzopyrenes and homologues were determined by high-resolution capillary gas chromatography (GC) with four different stationary phases: a 5% phenyl substituted methylpolysiloxane column (DB-5 ms), a 35% phenyl-substituted methylpolysiloxane column (BPX-35), a 50% phenyl-substituted methylpolysiloxane column (BPX-50), and a 35% trifluoropropylmethyl polysiloxane stationary phase (Rtx-200). Correlations for retention on each phase were investigated by using 8 independent molecular descriptors. Ri has been shown to be linearly correlated to PAH volume, polarisability alpha, Huckel-pi energy on the four examined columns. Ionisation potential Ip is a fourth variable which improves the regression model for DB-5ms, BPX-35, and BPX-50 column. Correlation coefficients ranging from r2 = 0.935 to r2 = 0.952 are then observed. Application of these indices to the identification and quantification of PAH with MW 302 in certified diesel particulate matter SRM 1650a is presented and discussed. PMID- 15335063 TI - GC-FID/MS method for the impurity profiling of synthetic d-allethrin. AB - A GC/FID/MS method was developed for the identification and quantification of d allethrin (DA) and its major impurities in commercial samples. Optimisation of the experimental conditions was carried out considering such important requirements as resolution, reproducibility, detection limits of 0.1% (m/m) for the impurities, and short analysis time. Under the optimised final conditions the method was validated for specificity, precision (CV% = 0.133 at 2.10 mg/mL and CV% = 0.035 at 3.00 mg/mL), linearity (0-3.00 microg injected), limits of detection (0.09 ng injected) and quantitation (0.28 ng injected), and robustness. The DA related impurities were identified by using a GC/MS method with ion trap mass detection and also by comparison with synthesised standards. The most abundant impurities were crysolactone, allethrolone, chrysanthemic acid, and chloro-derivatives of DA. PMID- 15335064 TI - Headspace solid-phase microextraction for characterization of fragrances of lemon verbena (Aloysia triphylla) by gas chromatography-mass spectrometry. AB - Natural fragrances from lemon verbena (Aloysia triphylla) were studied by headspace solid phase microextraction (HS-SPME) techniques followed by gas chromatography-mass spectrometry (GC-MS), with six different fibre coatings being tested to evaluate the extraction efficiencies of several selected compounds. A total of 14 compounds were identified in the fragrances of lemon verbena. Geranial and neral were detected as major components and alpha-pinene, beta pinene, beta-caryophyllene, and curcumene as minor components. Enantiomeric analysis of chiral compounds from lemon verbena was carried out on a chiral column. alpha-Pinene, limonene, and camphor in the fragrances emitted from lemon verbena were found in the (+), (-), and (-) forms, respectively. PMID- 15335065 TI - Characterization of cigarette smoke condensates by comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry (GC x GC/TOFMS). Part 2: basic fraction. AB - Cigarette smoke condensate is a complex chemical matrix. Analysis of nitrogen containing compounds present therein is very difficult because of the limitation of the peak capacity of conventional one-dimensional chromatography. Extensive and laborious sample preparation is frequently required or selective detectors are frequently used. In this study, the basic fraction of mainstream cigarette smoke condensate has been investigated by using comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry (GC x GC/TOFMS). Auto data processing by TOFMS software combined with manual identification was used to assign the components. 377 nitrogen-containing compounds, including 155 pyridine derivatives, 104 quinoline/isoquinoine derivatives, and 56 pyrazine derivatives were tentatively identified. By selecting appropriate unique masses and in the light of the component positions in the structured chromatogram, alkyl substituted pyridines, pyrazines, and quinolines/isoquinolines were separately shown and further validated. The peaks of eight individual positional isomers of two-carbon-substituted pyridines and thirteen positional isomers of methyl substituted quinolines/isoquinolines were further confirmed, based on linear incremental retention behavior in combination with TOFMS and the structured chromatogram of GC x GC. PMID- 15335066 TI - Quantification and stability of salbutamol in human urine. AB - A sensitive method for the quantification of free salbutamol in human urine is described. Sample clean up is performed using SPE on a mixed phase extraction column. Derivatisation is performed with N-methyl-N trimethylsilyltrifluoroacetamide (MSTFA) and the extract is analysed by GC-MS. The method was found to be suitable for use in the doping field, where a cut-off limit of 1 microg salbutamol/mL urine is set by the International Olympic Committee (IOC) and approved by the World Anti-Doping Agency (WADA). Above that value a doping violation occurs. In addition, the stability of salbutamol in human urine has been evaluated. PMID- 15335067 TI - Elution and preliminary structure-retention modeling of polar and ionic substances in supercritical fluid chromatography using volatile ammonium salts as mobile phase additives. AB - Supercritical fluid chromatography has primarily been applied to relatively nonpolar analytes, even when polar organic solvents are used as modifiers. Here, we show that low levels of volatile ammonium salts as mobile-phase additives allow the elution of polar and even ionic organic materials such as sulfonate salts, carboxylate salts, polyamines, and quaternary ammonium salts. Also, volatile ammonium salts are compatible with mass spectrometric detection, in contrast to other common additives. We have performed preliminary structure activity-relationship modeling for retention in the CO2/methanol/NH4OAc/Deltabond Cyano system. We have developed a three-descriptor model, where one descriptor, the "relative negative charged surface" explains over 61% of the variance in the retention values. We suggest that two mechanisms have the greatest influence on retention in this system. One is related to the presence of a volatile ammonium salt, the other is related to the ability of a molecule to "hide" its atom with the greatest partial negative charge. PMID- 15335068 TI - Capillary isoelectric focusing of proteins utilizing poly(vinylpyrrolidone)- and plexiglas-coated columns. AB - Fused-silica capillaries chemically derivatized with silane/poly(vinylpyrrolidone) (PVP) or dynamically modified with plexiglas [poly(methyl methacrylate)] were prepared and evaluated with regard to column stability and separation performance for capillary isoelectric focusing of standard proteins. The PVP coating showed the better stability and was good for at least 100 runs while the plexiglas coating started to deteriorate after about 30 runs. The time spent for the plexiglas coating is about 40 minutes while the PVP coating requires two days. The migration time reproducibility was better with the PVP capillary (RSD 0.7-1.6%, n = 5) compared to the plexiglas-coated column (RSD 1.2-2.9%, n = 5) while peak area and height varied over a similar interval (RSD 2-28.1% area; 0.9-22.7% height, n = 5). The two most consistent proteins in this evaluation, viz. myoglobin A and carbonic anhydrase II, showed linear dynamic ranges between 5-150 and 5-50 microg/mL, and limits of detections at 2 and 1 microg/mL, respectively, employing UV detection at 280 nm. PMID- 15335070 TI - 5th Balaton symposium on high-performance separation methods. PMID- 15335069 TI - Determination of betulin in Grewia tiliaefolia by HPTLC. AB - A sensitive, rapid, simple, and accurate high performance thin layer chromatographic method has been developed to standardize the bark of Grewia tiliaefolia Vahl. (Family: Tiliaceae) using betulin as an analytical marker. Chloroform extracts of bark from five different sources were used for HPTLC on silica gel with toluene-ethyl acetate, 90 + 10 v/v, as mobile phase. Under these conditions, the Rf of betulin was 0.22. The calibration plot was linear in the range of 1000 ng to 1800 ng of betulin and the correlation coefficient, 0.999, was indicative of good linear dependence of peak area on concentration. The mean assay of betulin was 2.596 +/- 0.594 mg g(-1) of bark. The method permits reliable quantification of betulin and good resolution and separation of betulin from other constituents of Grewia tiliaefolia. Recovery values from 96.09 to 98.87% showed that the reliability and reproducibility of the method were excellent. The proposed HPTLC method for quantitative monitoring of betulin in Grewia tiliaefolia can be used for routine quality testing. PMID- 15335071 TI - Comprehensive two-dimensional gas chromatography--GC x GC. PMID- 15335072 TI - The evolution of comprehensive two-dimensional gas chromatography (GC x GC). AB - For a technology little over a decade old, comprehensive two-dimensional gas chromatography (GC x GC) has quickly reached the status of one of the most powerful analytical tools for volatile organic compounds. At the heart of any GC x GC system is an interface, which physically connects the primary and the secondary columns and acts to preserve the separation obtained in the first dimension (first column) while allowing additional separation in the second dimension. The paper presents a review of the technology, including fundamental principles of the technique, data processing and interpretation and a timeline of inventive contributions to interface design. In addition, applications of the technique are presented, with a more detailed discussion of selected examples. PMID- 15335073 TI - Temperature requirements for thermal modulation in comprehensive two-dimensional gas chromatography. AB - Temperature requirements for trapping and release of compounds in a cryogenic gas loop-type GC x GC modulator were determined. Maximum trapping temperatures on the uncoated, deactivated modulator capillary were determined for compounds from C4 (bp -0.5 degrees C) to C40 (bp 522 degrees C). The liquid-nitrogen cooled gas flow rate was reduced from a high of 15.5 to 1.5 SLPM over the range to achieve the required trapping temperature. Excessive cold jet flow rates caused irreversible trapping and peak tailing for semi-volatile compounds above C26. Alternate cold jet coolants were investigated. An ice water-cooled jet was able to trap compounds with boiling points from C18 (bp 316 degrees C) to C40 and a room temperature air-cooled jet was able to trap compounds from C20 (bp 344 degrees C) to C40. The hot jet produced launch temperatures approximately 40 degrees C hotter than the elution temperature with heating time constants of 8 to 27 ms. Modulated compound peaks were symmetrical with half-height peak widths of 43 to 56 ms for compounds with little second column retention, and 70 to 75 ms for compounds with more second column retention. The liquid nitrogen-cooled loop modulator with gas flow programming was used to produce a GC x GC chromatogram for a crude oil that contained compounds from C7 to C47. PMID- 15335074 TI - A systematic analysis of a flow-switching modulator for comprehensive two dimensional gas chromatography. AB - A simple flow-switching device has been designed for use as a comprehensive two dimensional gas chromatography modulator. The device is constructed from fused silica tubing, t-unions, and a solenoid valve. A series of experiments were conducted to determine the influence of primary flow, secondary flow, modulation time, and device dimensions on the performance of the modulator. The flow switching device was found to produce pulses with widths near the theoretical minimum. High-performance was maintained over a wide range of modulation times. The flow-switching device did not introduce extra broadening along the primary retention axis. However, the modulator performance was optimal only over a narrow range of primary to secondary flow ratios. The ideal flow ratio is determined by the dimensions of the tubes that connect the t-unions. A simple flow resistance model has been developed that can predict the dimensions that will produce optimal results for a specified primary to secondary flow ratio. Thus, it is possible to construct a device that operates near the theoretical limit without numerous alterations. Under optimal conditions, the flow switching modulator generates peaks that are narrower than those produced by a diaphragm valve. PMID- 15335075 TI - Automated on-line comprehensive two-dimensional LC x GC and LC x GC-ToF MS: instrument design and application to edible oil and fat analysis. AB - After a successful off-line feasibility study, the automation of comprehensively coupled liquid chromatography and gas chromatography (LC x GC) has been studied. Important aspects to consider when developing automated LG x GC include the relative speeds of the two dimensions, the compatibility of the LC eluent (type and flow rate) with the GC dimension, and the column loadabilities. Because the GC separation is relatively slow, the LC instrument has to be operated in the stop-flow mode. Two interfaces for transferring large numbers of subsequent LC fractions to the GC were constructed: one based on a six-port switching valve, and one which uses a dual side-port syringe. Both interfaces were found to perform fully acceptably. The actual transfer of the LC fraction to the GC was realised using a standard split injector to vaporise the compounds and LC eluent. Gas phase splitting was applied to match LC mass load and GC column loadability. The standard deviations of the peak areas obtained in this way were better than 7% (n = 6). The reliability of the system was demonstrated by the problem-free analysis of large series of oil and fat samples, with the focus on both intact triglycerides and their fatty acid methyl esters (FAMEs). Finally, the hyphenation of the automated LC x GC system to a sensitive and rapid-scanning time-of-flight mass spectrometer was realised. By using LC x GC-ToF MS, the information density of the chromatograms could be improved even further, which allowed easy identification of individual compounds as well as compound groups. PMID- 15335076 TI - Quantification of naphthalenes in jet fuel with GC x GC/Tri-PLS and windowed rank minimization retention time alignment. AB - Comprehensive, two-dimensional gas chromatography (GC x GC) is used in conjunction with trilinear partial least squares (Tri-PLS) to quantify the percent weight of naphthalenes (two-ring aromatic compounds) in jet fuel samples. The increased peak capacity and selectivity of GC x GC makes the technique attractive for the rapid, and possibly less tedious analysis of jet fuel. The analysis of complex mixtures by GC x GC is further enhanced through the use of chemometric techniques, including those designed for use on 2-D data such as Tri PLS. Unfortunately, retention time variation, unless corrected, can be an impediment to chemometric analysis. Previous work has demonstrated that the effects of retention time variation can be mitigated in sub-regions of GC x GC chromatograms through the application of an objective retention time alignment algorithm based on rank minimization. Building upon this previous work, it is demonstrated here that the effects of retention time variation can be mitigated throughout an entire GC x GC chromatogram with an objective retention time alignment algorithm based on windowed rank minimization alignment. A significant decrease in calibration error is observed when the algorithm is applied to chromatograms prior to construction of Tri-PLS models. Fourteen jet fuel samples with known weight percentages of naphthalenes (ASTM D1840) were obtained. Each sample was subjected to five replicate five-minute GC x GC separations over a period of two days. A subset of nine samples spanning the range of weight percentages of naphthalenes was chosen as a calibration set and Tri-PLS calibration models were subsequently developed in order to predict the naphthalene content of the samples from the GC x GC chromatograms of the remaining five samples. Calibration models constructed from GC x GC chromatograms that were retention time corrected are shown to exhibit a root mean square error of prediction of roughly half that of calibration models constructed from uncorrected chromatograms. The error of prediction is lowered further to a value that nearly matches the uncertainty in the standard percent weight values (ca. 1% of the median percent volume value) when the aligned chromatograms are truncated to include only regions of the chromatogram populated by naphthalenes and compounds of similar polarity and boiling point. PMID- 15335077 TI - Assessment by monte carlo simulation of thermodynamic correlation of retention times in dual-column temperature programmed comprehensive two-dimensional gas chromatography. AB - The correlation of retention times in comprehensive two-dimensional gas chromatography caused by correlation of enthalpy and entropy changes between two stationary phases, methylsilicone and poly(ethylene glycol), was examined using commercial GC software and in-house Monte Carlo simulation. The enthalpy change, deltaH0, and entropy change, deltaS0, of 93 compounds were predicted from isothermal one-dimensional gas chromatograms predicted by the software. These values then were mimicked by Monte Carlo simulation, which removed the strong correlation of deltaH0 and modest correlation of deltaS0 between the two phases. Retention times in a comprehensive two-dimensional gas chromatogram (GC x GC) and in simulations of it were predicted for typical dual-capillary temperature programmed conditions using the actual, correlated values of deltaH0 and deltaS0 and their uncorrelated Monte Carlo counterparts, respectively. The uncorrelated deltaH0 and deltaS0 values caused the retention-time range of the simulations' second dimension to expand substantially beyond that in the GC x GC. Other simulations were developed using a restricted range of uncorrelated deltaH0 and deltaS0 values to mimic more closely the retention-time range of the GC x GC's second dimension. The intervals between nearest neighbor retention-time coordinates were calculated in both the latter simulations and the GC x GC. The intervals were larger in the simulations than in the GC x GC because the former contained uncorrelated coordinates and the latter contained correlated ones, which clustered along or near the diagonal. The retention times in the first dimension of the GC x GC were Poisson distributed, as assessed by statistical overlap theory. In contrast, the two-dimensional reduced retention-time coordinates in the GC x GC were not Poisson distributed, because retention times were correlated. However, the reduced retention-time coordinates in the simulations were Poisson distributed. PMID- 15335078 TI - Comprehensive two-dimensional gas chromatography in stop-flow mode. AB - The theory and proof of concept of a new mode of operation for comprehensive two dimensional gas chromatography (GC x GC) is presented. In current GC x GC interface designs, the modulation period defines the separation time allowed in the second dimension. In the stop-flow GC x GC mode, flow in the primary column is periodically stopped for brief periods of time. Consequently, the modulation period for the primary column and the amount of time available for second dimension separation become independent variables. This allows the separations in both the primary and secondary dimensions of the GC x GC system to be carried out under more optimised conditions, allowing extended periods of time for second dimension separations without sacrificing the separation in the primary dimension. This new technique has the potential to offer increased separation power and overall resolution as it is further developed. PMID- 15335079 TI - Comprehensive multidimensional GC for the characterization of roasted coffee beans. AB - The present investigation is based on the separation of one of the most complex food matrices: the roasted coffee bean volatile fraction. Analysis of the two main species of coffee (Arabica/Robusta) was achieved through an effective and simple sampling procedure, headspace solid-phase microextraction (SPME), and the unprecedented resolving power of comprehensive gas chromatography (GC x GC). The combination of these two techniques proved to be a powerful tool for the extraction and separation of coffee volatiles. In fact, thousands of compounds that play various roles in the constitution of coffee aroma profile were resolved in the 2-D contour plot, each occupying a specific position pinpointed by two retention time coordinates. The potential use of this method for the assessment of coffee quality and the detection of commercial fraud is discussed. The potential of GC x GC for identification and classification of unknowns was also demonstrated, as the formation of characteristic patterns for structurally related compounds was observed in the bidimensional chromatogram. Moreover, reproducibility results were supported by the use of an autosampler for SPME applications that allowed any inaccuracy arising from manual handling to be avoided. PMID- 15335080 TI - Application of headspace solid-phase microextraction (HS-SPME) and comprehensive two-dimensional gas chromatography (GC x GC) for the chemical profiling of volatile oils in complex herbal mixtures. AB - The coupling of headspace solid-phase microextraction (HS-SPME) with comprehensive two-dimensional gas chromatography (GC x GC) was shown to be a powerful technique for the rapid sampling and analysis of volatile oils in complex herbal materials. When compared to one-dimensional (1-D) GC, the improved analytical capabilities of GC x GC in terms of increased detection sensitivity and separation power were demonstrated by using HS-SPME/GC x GC for the chemical profiling (fingerprinting) of essential/volatile oils contained in herbal materials of increasing analytical complexity. More than 20 marker compounds belonging to Panax quinquefolius (American ginseng) can be observed within the 2 D contour plots of ginseng itself, a mixture of ginseng and another important herb (P. quinquefolius/Radix angelicae sinensis), as well as a mixture of ginseng and three other herbs (P. quinquefolius /R. angelicae sinensis/R. astragali/R. rehmanniae preparata). Such analytical capabilities should be important towards the authentication and quality control of herbal products, which are receiving increasing attention as alternative medicines worldwide. In particular, the presence of Panax in the herb formulation could be readily identified through its specific peak pattern in the 2-D GC x GC plot. PMID- 15335081 TI - Comprehensive two-dimensional gas chromatography in the analysis of dietary fatty acids. AB - Comprehensive two-dimensional gas chromatography utilising a semi-rotating cryogenic modulator was applied to the analysis of dietary milk derived fatty acids. Four column combinations were tested: two polar-nonpolar and two different nonpolarpolar column combinations. Best results were obtained with a nonpolar polar column set, with narrow-bore (50 microm ID) Carbowax column as the second column. The GC x GC method clearly improved the separation efficiency of the fatty acids in comparison with one-dimensional GC analysis and proved to be suitable for the quantification of the analytes. The relative standard deviations (RSD) of the retention times in the first and second dimensions were on average 0.06% and 4.9%, respectively. The relative RSD for peak volumes was on average 2.9%. PMID- 15335082 TI - Speciation of nitrogen-containing compounds in diesel fuel by comprehensive two dimensional gas chromatography. AB - Nitrogen-containing compounds in diesel fuel have been speciated by comprehensive two-dimensional gas chromatography with nitrogen chemiluminescence detector (GC x GC-NCD). The speciation of nitrogen-containing compounds in diesel is difficult because of low concentration and complexity. The advantages of GC x GC are improved resolution and enhanced sensitivity. GC x GC-NCD can achieve the type and class separation of nitrogen-containing compounds with an appropriate separation column combination. Diesel contains both neutral (indoles and carbazoles) and basic (pyridines and quinolines) nitrogen-containing compounds. Relative concentrations of each class as well as each carbon number family can be quantified by integrating their peak volumes. This study demonstrates the capability of GC x GC-NCD for speciation of nitrogen-containing compound classes. PMID- 15335083 TI - Application of comprehensive multidimensional gas chromatography combined with time-of-flight mass spectrometry (GC x GC-TOFMS) for high resolution analysis of hop essential oil. AB - The selection and quality of hops is a major determinant in beer flavour. Brewers acknowledge that distinctive characteristics of different hop varieties can be traced to the composition of their essential oils. The difficulty in characterising complex mixtures such as hop oil using 1-D chromatography is that many compounds co-elute. With the introduction of comprehensive multidimensional capillary gas chromatography (GC x GC), there is a tremendous improvement in the separation power or peak capacity. Recent work using GC x GC with flame ionisation detection has suggested that there may be over 1,000 compounds in hop oil. This work describes the use of GC x GC combined with TOFMS detection (Leco Pegasus 4D instrument) to analyse Target hop oil. The TOFMS spectral acquisition rate of 60 Hz provided sufficient spectra per peak (2-D peak base width of 0.1 0.2 s) for identification (119 components were identified with 45 previously unreported compounds). When analysing results, an advantage of GC x GC coupled to TOFMS is that 2-D chromatograms can be viewed for individual masses that are characteristic of particular functional groups. This allows the analyst to view the various homologous series of compounds although in certain cases coelution may still be present as shown by the esters with mass 75. PMID- 15335084 TI - 2004 ACS Award in Separation Science and Technology to William H. Pirkle. PMID- 15335085 TI - Shigeru Terabe receives the 2004 ACS Award in Chromatography. PMID- 15335086 TI - Patient maintained propofol sedation for dental surgery. AB - AIM: This study evaluated a system for patients to operate a target-controlled infusion (TCI) of propofol for sedation. PARTICIPANTS: 50 healthy adults, undergoing minor oral surgical procedures under local anaesthesia. METHODS: Following instructions, patients were allowed to control their own sedation to comfortably undergo surgery. OUTCOME MEASURES: Operating conditions, degree of sedation, vital signs and patients' assessment of the technique. RESULTS: Operating conditions were described as good in 47 patients and fair in 3. Optimal sedation was provided at a median target blood concentration of 2 mcg/ml (range1 3mcg/ml). 30 were sedated moderately, 16 deeply and 4 mildly. None were oversedated. The vital signs were stable in all patients. 30% of patients had injection pain and 28% became talkative. Patient satisfaction was high with 86% feeling adequately relaxed and 92% willing to use it again. 14 patients had complete and 22 had partial amnesia for the procedure. CONCLUSIONS: This technique combines the benefits of target-controlled infusion with patient controlled feedback and produces safe dental sedation. PMID- 15335087 TI - Autoclave use in dental practice in the Republic of Ireland. AB - AIMS: To assess by postal questionnaire, cross-infection control methods, especially sterilisation procedures, of 700 general dental practitioners in the Republic of Ireland, and to biologically monitor steam pressure sterilisers or autoclaves in their practices. MAIN OUTCOME MEASURES: Methods of instrument cleaning and sterilisation, autoclave efficacy. RESULTS: A response rate of 40% with all, except one practitioner, using steam sterilisation. 49% also reported the use of chemical sterilisation with a quarter of these using glutaraldehyde. However, instrument soaking time varied greatly from 2.5 minutes to 74 hours. Methods of instrument cleaning prior to autoclaving were as follows: scrubbing by hand 41.5%, ultrasonic cleaning 7.0%, combination of both 50%. 52.9% of the respondents did not autoclave their dental handpieces and only 44.7% disinfected impressions before sending them to the laboratory. The autoclaves of thirty practitioners (11.3%) did not pass the initial biological test. Following counselling about possible causes of failure, four autoclaves (1.5%) failed a repeat biological test. However, seven practitioners did not return the repeat biological test. CONCLUSIONS: Some aspects of recommended cross-infection control procedures are well adhered to, e.g. instrument cleaning, but further education is required in certain key areas, in particular the use of chemical sterilisation, dental handpiece autoclaving and impression disinfection. There is also a need to increase awareness of the importance of routine autoclave servicing and calibration, along with validation and monitoring. PMID- 15335088 TI - Influence of sociodemographic variables on use of dental services, oral health and oral hygiene among Spanish children. AB - OBJECTIVES: To analyse the possible influence of sociodemographic variables on use of dental services, oral health and oral hygiene among Spanish children. METHODS: Descriptive cross-sectional study based on 1,676 interviews conducted with parents or guardians of children ages 3-15 years. The dependent variables analysed were: use of dental services in the preceding 12 months; self-reported caries ever; and daily dental hygiene measured as toothbrushing frequency. Independent variables were sex, age, size of town or city, educational level, and monthly income of the family unit. RESULTS: A total of 60% of the study children had not visited a dental professional in the preceding 12 months. Children having parents or guardians with the lowest educational level were 1.592 times more likely to have received no dental care than those with the highest educational level. The likelihood of not having made use of such dental services rose 2.232 fold in cases where income was < 600 versus >1200 per month. Overall prevalence of self-reported caries for the sample as a whole was 34.4%. Subjects in the lowest income bracket (< 600 per month) were 1.497 times more likely to have caries than those in the highest income bracket. In all, 68.5% brushed their teeth every night, but the percentage rose significantly with age, size of town or city, and income. CONCLUSION: There is considerable social inequality in the use of dental services, oral health and oral hygiene among Spanish children. PMID- 15335089 TI - Caries prevalence 1987-1998 in 12-year-olds in France. AB - OBJECTIVE: To describe caries morbidity in 12-year-old French schoolchildren 1987 1998. MATERIALS AND METHODS: Three cross-sectional surveys from the WHO Global Oral Data Bank that were conducted at national level in France in 1987, 1993, 1998. The surveys involved a representative sample of 1,000 12-year old children in 1987, 1,331 in 1993, and 6,000 in 1998. RESULTS: An important decline of the caries focused in the first period - 22.8% vs. 4.5% in the period 1993-1998. All components in the DMFT Index decreased although they followed different patterns. The mean number of filled teeth consistently dropped over the entire 11 years in a context where the mean number of untreated carious teeth reached its lowest point in 1993 and increased thereafter. The four first molars accounted for the vast majority of the caries, 73.7% in 1998. In 1998 year, 44.9 % of children had caries-free first molars as opposed to 38.8 % in 1993 and 14.0 % in 1987. CONCLUSION: The influence of this caries evolution on the oral health system is discussed. Suggestions are advanced to integrate dentistry in a public health concept especially focused on the reduction of inequality. PMID- 15335090 TI - Single tooth crowns supported on hydroxyapatite coated endosseous dental implants: a prospective 5-year study on twenty subjects. AB - AIM: To evaluate the success and maintenance requirements of cylindrical, hydroxyapatite coated implants used as single-tooth implants. DESIGN: A five-year prospective trial. METHOD: Twenty patients, attending the Department of Prosthetic Dentistry at GKT Dental Institute, London, were provided with 23 Calcitek, Integral Omniloc single tooth implants. Annual reviews were undertaken for five years. Assessment criteria included implant survival; abutment or crown looseness; radiographic evidence of bone loss; evidence of plaque and inflammation of the mucosal cuff; the patient's perception of the restoration. RESULTS: All implants integrated. Two subjects, with three crowns, dropped out giving a known cumulative survival rate of 87%. Twelve implants exhibited no measurable bone loss and six demonstrated funnel formation up to 2mm in depth which, at five years, appeared to be stable; three implants exhibited continuing bone loss and their inclusion as failures gives a known success rate of 74%. The abutments came loose on four occasions; the crowns decemented on 12 occasions; two crowns were repaired and one remade. The appearance was judged to be good or very good. CONCLUSION: The prospective study indicated that hydroxyapatite coated implants can be successful as single tooth implants, over a five-year period. PMID- 15335091 TI - Prevalence of pulpally involved permanent teeth in Saudi schoolchildren. AB - OBJECTIVE: To determine the prevalence of pulpally involved permanent teeth in children and adolescents aged 6-18 years in Saudi Arabia. METHOD: A systematic random sample was collected from the primary care files in the College of Dentistry in Riyadh. Data were collected from a total of 997 files and entered into a database and statistical program. RESULTS: A high prevalence rate of 35.8% of individuals with pulpally involved teeth was found. More extractions, pulp cappings, and root canal treatments were provided as the age increased. In all age groups molars were the most affected, and canines were the least. Maxillary incisors were affected more than mandibular incisors, while mandibular molars were affected more than maxillary molars. Females had more pulpally involved teeth than males. CONCLUSIONS: It is suggested that early oral interventions, prevention and dental education for the patients and their families may decrease this high prevalence level of pulpal involvement in teeth in this age group. PMID- 15335092 TI - Effect of home bleaching agents on the roughness and surface morphology of human enamel and dentine. AB - AIMS: Carbamide peroxide (CP) containing bleaching agents are now widely used in different concentrations. In this study, the effect of 10% and 15% CP containing home bleaching agents on the surface roughness and morphology of human enamel and dentine were evaluated in vitro. METHODS: 90 non-carious human mandibular incisors were divided randomly into two equal groups (enamel and dentine). The dentine group specimens were ground to expose dentine surfaces. The samples in both groups were then randomly divided into three main groups and then divided into three subgroups, according to bleaching application times (n=15). The samples in both enamel and dentine groups were treated with 10% CP at Group 1 and 4, 15% CP at Group 2 and 5 and distilled water at Group 3 and 6. The surface roughness of the samples was analysed before bleaching (1a, 2a, 3a for enamel, 4a, 5a, 6a for dentine), after 4hr (1b, 2b, 3b for enamel, 4b, 5b, 6b for dentine) and after 28hr of bleaching application (1c, 2c, 3c for enamel, 4c, 5c, 6c for dentine) using a computerised roughness tester. The surface morphologies of treated and control specimens were also examined by SEM. RESULTS: There were no statistically significant differences between the surface roughness of untreated control specimens and the specimens treated with the bleaching materials (10% and 15% CP) for both enamel and dentine at any given measurement time (p>0.05). CONCLUSION: Under the careful guidance of a dentist, home bleaching can be thought a reliable treatment for both stained enamel and dentine, regardless of whether 10% or 15% CP is used. PMID- 15335093 TI - The effect of two different dental gels and a mouthwash on plaque and gingival scores: a six-week clinical study. AB - AIM: To evaluate the clinical efficacy of two gel formulations containing chlorhexidine gluconate and neem extract with a commercially available chlorhexidine gluconate mouthwash. METHOD: An open labelled randomised six-week clinical study with parallel group design in 48 subjects divided into four groups. Plaque accumulation and gingival condition were recorded using plaque index and gingival index. On the basis of mean baseline plaque and gingival scores, subjects were allocated to four different groups, using their assigned products twice a day, before bed and after breakfast. Plaque and gingival scores were recorded after three and six weeks. RESULTS: Mean plaque and gingival scores were reduced over the six-week trial period for experimental and control groups. Chlorhexidine gluconate gel reduced the plaque and gingival scores significantly more (p<0.05) than the chlorhexidine gluconate mouthwash. Neem extract gel also showed significant (p<0,05) reduction in plaque and gingival scores when compared with the control group. But there was no significant difference between the groups treated with chlorhexidine gel and neem extract gel. CONCLUSION: The results of this clinical study indicate that better therapeutic efficacy can be achieved using gels for treating oral infections than conventional treatments using mouthwash. PMID- 15335094 TI - A review of satisfaction with dental service at a Jerusalem community clinic serving elderly patients. AB - AIM: The audit of dental satisfaction and its association with the reported outcome of a dental health care programme for elderly patients. DESIGN: A telephone survey of 162 patients in a Jerusalem municipal dental clinic for geriatric patients, who had completed a treatment course in oral rehabilitation. RESULTS: The average age of the respondents was 73.2 +/- 8.45 years, over 80% of who were very satisfied or satisfied with the treatment given by the dental clinic staff. Only 1.8% were not satisfied (the remaining 17.9% were unable to answer the question). Among those who responded, 91.6% were currently using their full dentures, 80.4% reported an improvement in chewing ability, and 82.3% an improvement in appearance. Respondents who reported daily use of dentures and improved chewing and appearance, also reported higher levels of satisfaction with the clinic. CONCLUSIONS: These results indicate a potentially important role of satisfaction with dental staff in optimal compliance and success of a geriatric oral health care programme. PMID- 15335095 TI - Knowledge matters. PMID- 15335096 TI - Gene profiling--chances and challenges. AB - Microarray analysis has been emerged as a tool to characterize the overall reaction of cells in culture or tissue to different stimuli e.g. stressful events by analysing bulk RNA present at a particular time point. It has supplemented or even replaced more traditional methods like cDNA-bank sequencing or conventional differential display. The commercial availability of several different precoated arrays and the ease of handling has supported the broad distribution of this new technique. The basic protocol involves the hybridization of complementary strands of labelled DNA or RNA from cells/tissue with representations of known genes spotted onto a solid support (nylon, glass). Labelling can be radioactive (p32/33), by a hapten group (biotin, digoxigenin, aminoallyl) or by fluorescent (Cy3, Cy5 etc.) nucleotides. Detection is performed by autoradiography, chemiluminescence or fluorescence scanning. There are different setups of arrays available: either known genes/gene-groups (apoptosis, cytokines etc.) are spotted as PCR fragments, plasmids or synthetic oligonucleotides or representations of the known genome are directly synthesized as short sequence tags of 20-70 oligonucleotides on glass chips. The latter allow the identification of newly expressed genes whereas the former deal with known genes. Ideally, the intensity of the signal can be correlated with the relative expression of a known gene and allows the comparison with a standard. Problems arise from the quality of the sample material, the standardization of the protocols and the data management. Nevertheless, gene profiling by cDNA-arrays will definitely be integrated into routine screening programs. PMID- 15335097 TI - Role of mitochondrial proteins for neuronal cell death after focal cerebral ischemia. AB - Cell death following focal cerebral ischemia has an acute and a delayed component. Delayed neuronal cell death occurs via activation of molecular signalling pathways resembling apoptosis in nonneuronal cells. Cell surface cell death receptors and damage to mitochondria or DNA initiate these pathways finally leading to DNA fragmentation and cell death. Central mediators of delayed neuronal cell death are two families of molecules: a group of cysteine aspartate proteases, called caspases, and molecules of the bcl-2 family, e.g. bcl-2, bax, and bid. Caspases initiate and execute cell death, while bcl-2 family members modulate death signalling and lead to release of pro-apoptotic molecules from the mitochondrial intermembranous space, e.g. cytochrome c and apoptosis inducing factor (AIF). Cytochrome c induces cell death by activation of caspase 9 and 3, while AIF leads to detrimental DNA damage by an capase-independent pathway. The current paper reviews recent findings dealing with pre- and post-mitochondrial cell death pathways activated by focal cerebral ischemia. PMID- 15335098 TI - Signalling mechanisms for survival of lesioned motoneurons. AB - Mechanisms controlling neuronal survival play an important role both during development and after birth, in particular when the nervous system is lesioned. Isolated embryonic motoneurons and other types of primary neurons have been a useful tool for studying basic mechanisms underlying neuronal cell death during development and under pathophysiological conditions after neurotrauma. These studies have led to the identification of neurotrophic factors which under physiological conditions regulate survival and functional properties, and after neurotrauma promote regeneration and plasticity. Functional analysis of these molecules, in particular by generation of gene knockout mice, has led to a more detailed understanding of complex requirements of individual types of neurons for their survival and also paved the way for a better understanding of the signalling pathways in lesioned neurons which decide on cell death or survival after axotomy and other pathophysiological conditions. These findings could ultimately lead to a rational basis for therapeutic approaches aiming at improving neuronal survival and regeneration after neurotrauma. PMID- 15335099 TI - Genetically modified animals in molecular stroke research. PMID- 15335100 TI - The Janus face of inflammation in ischemic brain injury. AB - Brain ischemia elicits an intense inflammatory reaction as evidenced by endogenous activation of microglia and infiltration of leukocytes from the systemic circulation into the brain. A key issue regarding the well-described inflammation in brain injury is whether this reaction is of salutary or detrimental nature in the short and longterm post ischemia. In this brief review, evidence in support for the possible beneficial as well as detrimental role of inflammatory cells and mediators in ischemic brain injury is highlighted. We offer the opinion that both benefits and adverse effects of the inflammatory reaction at large depent on the levels of a specific mediator, the temporal relationships to the injury, the context in which the mediator operates and the spatial relationships to the injury. PMID- 15335101 TI - The natural course of lesion development in brain ischemia. AB - Histopathologic and NMR imaging studies show that focally ischemic brain lesions tend to increase in size over time. In animal models of stroke as well as in patients presenting with hemispheric stroke, considerable lesion growth was observed. In focal cerebral ischemia, lesions predominantly enlarge early (<12 hrs postinsult) and show complete ischemic injury due to pan necrosis in the vast majority of affected tissue. In global cerebral ischemia--a condition that is present after cardiac arrest--lesions appear late (>12 hrs) in selectively vulnerable brain regions and neurons are damaged by apoptosis. The short resuscitation time of the brain explains why periods of global ischemia result in widespread and global loss of energy metabolites combined with diffuse brain edema and global damage. Mechanisms involved in lesion growth include excitotoxicity, peri-infarct depolarizations, lactacidosis, microcirculatory disturbances, and flow-metabolism uncoupling among others. Problems involved in the subject under focus are related to maturation phenomena of injury and the different imaging modalities (metabolic imaging, NMR imaging, positron emission tomography) that require a subtly differentiated interpretation of the alterations observed. PMID- 15335102 TI - Pharmacological preconditioning in global cerebral ischemia. AB - Single dose 3-nitropropionic acid (3-NPA) 24 hr before global ischemia improves neuronal survival in both, neocortex and hippocampus ('chemical preconditioning'). Neuronal survival after transient global ischemia requires new protein synthesis during recovery, especially of those with anti-apoptotic function. Bcl-2-protein is expressed in neurons that survive cerebral ischemia and may parallel the time course of tolerance after ischemic preconditioning. With this study we examined whether differences in bcl-2-protein expression compared to baseline may be involved in the induction of ischemic tolerance using 3-NPA. Male Wistar rats received either a single intraperitoneal (i.p.) dose of 3 NPA (20 mg/kg), and were observed for 3 (n = 4), 12 (n = 5) or 24 hours (n = 5) or the same amount of vehicle and were observed for 24 h (n = 8, controls). Immunohistochemistry allowed to compare the intensity of bcl-2 immunoreactivity at three subsequent time points in hippocampus, dentate gyrus and parietal neocortex with that of control animals. A single dose of 3-NPA caused a significant increase of bcl-2 protein immunoreactivity in hippocampal neurons, i.e. CA 1 (5 out of 5 animals, p = 0.003), CA 3 (5/5, p = 0.003), CA 4 (4/5, p = 0.025), and neocortex (5/5, p = 0.004), in a time dependent manner over a period of 24 hr after injection. Neuronal bcl-2 protein expression in CA 2 and dentate gyrus remained unchanged. The data suggest a possible role of bcl-2-protein in chemical induction of ischemic tolerance using a single subtoxic dose of 3-NPA. Bcl-2-protein expression may be initiated by increased levels of reactive oxygen species (ROS) after 3-NPA administration, as shown by others. Additional bcl-2 protein may then be available to (1) control postischemic ROS burst, (2) protect the mitochondrial membranes, and (3) inhibit pro-apoptotic mechanisms. PMID- 15335103 TI - The role of inflammatory processes in the pathophysiology and treatment of brain and spinal cord trauma. AB - Traumatic injury to the brain and spinal cord results in an early inflammatory response that is initiated by the release of proinflammatory cytokines followed by the infiltration and accumulation of polymorphonuclear leukocytes (PMNLs). The role of the inflammatory cascade on traumatic outcome remains controversial. Pleiotropic cytokines appear to function both protectively and destructively. The induction of cytokines can lead to the expression of the inducible form of nitric oxide synthase (iNOS), which in turn provokes the release of excessive amounts of nitric oxide (NO) that may participate in the pathogenesis of tissue injury. Hypothermia has been reported by various groups to be neuroprotective in brain and spinal cord trauma. We studied the effect of therapeutic hypothermia on cerebral IL-1beta concentrations, PMNL accumulation and iNOS activity after traumatic brain injury (TBI) and spinal cord injury (SCI). Based on current data therapeutic hypothermia may protect in models of traumatic injury by modulating deleterious inflammatory processes. PMID- 15335104 TI - L-type calcium channel antagonist nifedipine reduces neurofilament restitution following traumatic optic nerve injury. AB - BACKGROUND: The aim of the present study was to observe if the use of the L-type calcium channel antagonist nifedipine would offer advantages for the retinal ganglion cells and the restitution of the axonal cytoskeleton after optic nerve crush. METHODS: Retinal ganglion cells were retrogradely labeled with a fluorescent calcium marker. With the in vivo confocal neuroimaging (ICON) method we observed the fluorescent cell metabolism marker Oregon Green BAPTA in the same retinal ganglion cells over 3 weeks after optic nerve crush. 2 micromol nifedipine were injected intraocularly 30 minutes following optic nerve crush. FINDINGS: Investigation of the optic nerve immunostained for NF-H presented decreased restitution of the neurofilaments in the axonal cytoskeleton after 3 weeks in the optic nerve crush group treated with nifedipine as compared to the optic nerve crush only group. INTERPRETATION: These results show that a single injection of the calcium L-type antagonist nifedipine shortly after optic nerve injury has long-lasting negative effects on the recovery of the retinal ganglion cells. PMID- 15335105 TI - Neuronal activity and cortical perfusion determined by quantitative EEG analysis and laser doppler flowmetry are uncoupled in brain injured rats. AB - Following traumatic brain injury uncoupling of metabolism from perfusion, reflected by an increase in cellular activity in face of reduced perfusion contributes to secondary tissue damage. Standard autoradiographic techniques employed to study metabolism and perfusion are limited by the inability to perform longitudinal investigations. Therefore, the aims of this study were to investigate if metabolic uncoupling can be studied by applying noninvasive and non-radioactive methods. For this, quantitative EEG analysis and laser Doppler flowmetry were employed to determine changes in EEG activity as a global measure of neuronal activity and pericontusional cortical perfusion during the early phase following controlled cortical impact injury (CCII). In eight rats changes in neuronal activity and pericontusional cortical perfusion were determined before, at 4 and 24 hours after CCII. Neuronal activity was significantly increased by 40% at 4 hours after CCII followed by a significant decrease by 60% at 24 hours compared to pre-trauma levels. Pericontusional cortical perfusion was significantly reduced by 45% and 18% at 4 and 24 hours after CCII. respectively. Non-radioactive methods like quantitative EEG analysis and laser Doppler flowmetry can be used to reveal underlying uncoupling of EEG activity from cortical perfusion which is mostly sustained in the early phase following CCII. PMID- 15335106 TI - Glial scar and axonal regeneration in the CNS: lessons from GFAP and vimentin transgenic mice. AB - Astrocytes play an active role in the brain and spinal cord. For example, they have a function in formation and maintenance of the blood-brain barrier, ion homeostasis, neurotransmitter transport, production of extracellular matrix, and neuromodulation. Moreover, they play a role in preserving or even restoring the structural and physiological integrity after tissue injury. Currently, the function of astrocytes was studied with regard to the controversially discussed aspects of permissivity on the one-hand-side and inhibition of the other side exerted by reactive astrocytes for axonal regrowth in the adult CNS. Accordingly, knock-out mice deficient in vimentin (VIM) and/or glial fibrillary acidic protein (GFAP), the two major IF-proteins of astrocytes, were investigated. In addition, in vitro studies were carried out, on whether the absence of one or both proteins (VIM, GFAP) influences axonal regeneration. In experimental animals, a hemisection of the spinal cord was performed utilizing the above mentioned double mutant mice. The knock-out mice were generated by gene targeting. Double-mutants were obtained by crossing single null mice. The in vitro results indicate that both VIM and GFAP were absent in astrocytic cultures obtained from double-mutant mice. On the other side, the proteins were detected in more than 85%, of cultured cells from wild types. Co-culture of mutant mice astrocytes with neurons revealed that the neuronal density was different from that obtained in culture with wild type astrocytes. On the other side, there was a marked increase in neuronal density in co-cultures utilizing both GFAP knock-out- or double-mutant mice astrocytes again as compared to co-cultures with wild type astrocytes. Moreover, the neurite length of neurons was significantly increased in experiments with neurons growing on astrocytes from GFAP-knock-out or double-mutant mice. The in vivo experiments demonstrate an increase of nestin (NES) immunoreactivity at three days in the sectioned side of the spinal cord, in the perikaryon and astroglial processes. In double-mutant mice only a slight increase in NES immunoreactivity was found in the lesion side, albeit confined to the perikaryon of astrocytes. Below the lesion, serotonin immunostaining was dramatically reduced three days after the insult in both sides, particularly in the lesion side. The decrease was more pronounced in double-mutant than in wild type mice. On the other side, double-mutant mice had a much higher density of serotonergic fibers in the ventral horn in the lesioned side. In conclusion, the findings demonstrate that in the absence of important astrocytic proteins as VIM and GFAP, the astroglial response to injury is significantly modified underlying reduced scar formation. Attenuation of scar formation may enhance axonal sprouting of serotonergic axons below the lesion, which specifically reinnervate motoneuron pools. PMID- 15335107 TI - Recovery from spinal cord injury--underlying mechanisms and efficacy of rehabilitation. AB - Patients with an acute complete spinal cord injury (SCI) present a syndrome called "spinal shock". During spinal shock the loss of tendon reflexes and flaccid muscle tone is associated with a low persistence of F-waves and loss of flexor reflexes while H-reflexes are well elicitable. When clinical signs of spasticity become established, the electrophysiological parameters show little change. The divergent course of clinical signs of spasticity in their possible neuronal correlates indicate the occurrence of non-neuronal changes contributing to spasticity. - When signs of spinal shock had disappeared in patients with incomplete and complete paraplegia a locomotor pattern can be induced and trained under conditions of body unlaoding using a moving treadmill. In complete and incomplete paraplegic patients an increase of gastrocnemius electromyographic activity occurs during the stance phase of a step cycle with a daily locomotor training, coincident with a significant decrease of body unloading. In contrast to this, neither clinical nor electrophysiological examination scores improve. The locomotor pattern depends on the level of lesion: the higher the level of spinal cord lesion the more 'normal' is the locomotor pattern. This suggests that neuronal circuits underlying 'locomotor pattern generation' in man is not restricted to any specific level of the spinal cord, but extends from thoraco lumbal to cervical levels. PMID- 15335108 TI - Lessons from epidemiologic studies in clinical trials of traumatic brain injury. AB - Lessions from epidemiological studies. The Clinical Trial Group for Neurosurgery of the University of California San Diego (UCSD) is involved in epidemiological studies and trials of new pharmacological agents in traumatic brain injury. A great number (> 10,000) of patients has been prospectively analyzed forming an integrated database for further purposes. The development of these databases is based on earlier work by the European Neurosurgeons Jennett and Braakmann and the US-Traumatic Coma Data Bank Study. These studies allowed for the development of sophisticated data collection instruments which were used in the international Tirilizad Trials which enrolled over 1,100 patients. A major observation from that trial was that pretreatment hypotension or hypoxia could be unbalanced even in a large two arm blinded study. Another issue of the tirilazad trial was the influence of gender affecting not only outcome but also drug metabolism. Similar experiences were gathered with the phase-III trial on the competitive NMDA receptor antagonist selfotel, which interferes with the excitotoxic amino acid glutamate as mediator of secondary brain damage, as ischemia-induced neuronal degeneration. Unfortunately, the trial, already underway, had to be prematurely aborted, since concurrent stroke studies with enrollment of nonintubated patients on low-dose selfotel revealed an increased number of deaths and other adverse events. A retrospective analysis did not confirm that Selfotel was associated with an increased mortality in TBI, but there was also no evidence that the drug was efficacious. A problem here was that a major portion of patients did not have intracranial mass lesions (contusion, subdural haematoma) on CT, questioning whether these had a treatment responsive brain injury. Both studies on tirilazad or selfotel underscore the significance of well designed and conducted phase-I and -II trials to characterize the pharmacokinetics of the agent, to confirm availability of drug in the brain, and to identify a sufficient number of patients with lesions responding to the drug. A major issue is the blood-brain barrier permeability of the agent under study. Further, the phenomenon of secondary deterioration - neurological worsening - turned out as a powerful predictor of poor outcome. The findings and conclusions of both clinical trials (tirilazad, selfotel) were utilized for a subsequent patient study on CP101-606 in consultation with the Pfizer company, the US Brain Injury Consortium, and the San Diego Clinical Trial Group. The patient population was a priori selected towards responsiveness of the brain lesions to the treatment. The major conclusions are: I Development of therapeutic regimens targeted towards the mechanisms of brain injury. II Availability of adequate preclinical data. III Directing treatment towards an appropriate patient population. IV Central gathering and interpretation of the neuroradiological findings. V Monitoring of trial center performance. VI Stratification and pre-trial prognostic analysis for identification of subgroups. PMID- 15335109 TI - The MRC CRASH trial--a large, simple randomised trial of steroids in head injury. AB - CRASH (Corticosteroid randomisation after significant head injury) is a prospective multi-centre randomised double blind study of methylprednisolone versus placebo in mild, moderate and severe head injury. Patients are eligible up to 8 hours from injury. To date the CRASH trial has recruited 9000 patients. The trial is recruiting from 200 hospitals in 50 countries with another 100 centres planning to join the trial. The target for recruitment is 20,000 patients by 2006. The trial is wholly funded by the Medical Research Council of Great Britain and is multidisciplinary, involving doctors and nurses from a range of specialities. A recent systematic review of corticosteroids in head injury demonstrated a risk of death in the corticosteroid treated group 2% lower than in the control group. The 95% confidence interval ranges from a 60%, lower mortality to a 2% higher mortality. This result is compatible with there being no real benefit, but it is also compatible with there being a small benefit of a few percent. An improvement in mortality of 2% would theoretically save 10,000 lives per 500,000 patients treated. The global impact of such a treatment effect would be significant as the number of head injuries world-wide continues to rise. PMID- 15335110 TI - Clinical trials in traumatic brain injury: current problems and future solutions. AB - Over the past decade many neuroprotective agents have been developed with the hope of being able to improve outcome in patients with traumatic brain injury. Unfortunately, none of the phase III trials performed have convincingly demonstrated efficacy in the overall population. A common misconception is that consequently these agents are ineffective. Such has not been proven and some trials show evidence of efficacy in subgroups of the population studied. The negative results, as reported in the overall population, may in part be caused by specific aspects of the TBI population, as well as by aspects of clinical trial design and analysis. Clinical trials in TBI pose several complicated design issues. Methodological challenges relate particularly to heterogeneity of the population and to outcome assessment. Heterogeneity pertains both to the range of pathologies included in TBI, and to prognostic factors, each causing specific problems. Mechanistic and/or prognostic targeting, as well as possibilities for covariate adjustment, are suggested as possible solutions to deal with the problems of heterogeneity. The aim in most trials was to demonstrate a 10% absolute improvement in favorable outcome in patients with head injury. This may be considered overoptimistic and unrealistic in relation to the heterogeneous patient population. Specific problems are further incurred by the use of the dichotomized Glasgow Outcome Scale as primary outcome measure. Optimal statistical power may expected to be present when the point of dichotomization results in a 50:50 distribution of outcome categories. It is proposed to differentiate the point of dichotomization according to prognostic risk profile, in order to maintain statistical power. Solutions described may be expected to enhance chances of demonstrating benefit of potentially effective neuroprotective agents in future studies. The complexity of problems occurring in clinical trial design and analysis in TBI is such that a strong and sustained multidisciplinary input and effort is required from all experts involved in the field of neurotrauma. PMID- 15335111 TI - Prospective documentation and analysis of the pre- and early clinical management in severe head injury in southern Bavaria at a population based level. AB - Treatment of patients suffering from severe head injury is so far restricted to general procedures, whereas specific pharmacological agents of neuroprotection including hypothermia have not been found to improve the outcome in clinical trials. Albeit effective, symptomatic measures of the preclinical rescue of patients (i.e. stabilization or reestablishment of the circulatory and respiratory system) or of the early clinical care (e.g. prompt diagnosis and treatment of an intracranial space occupying mass, maintenance of a competent circulatory and respiratory system, and others) by and large constitute the current treatment based on considerable organizational and logistical efforts. These and other components of the head injury treatment are certainly worthwhile of a systematic analysis as to their efficacy or remaining deficiencies, respectively. Deficits could be associated with delays of providing preclinical rescue procedures (e.g. until intubation of the patient or administration of fluid). Delays could also be associated in the hospital with the diagnostic establishment of intracranial lesions requiring prompt neurosurgical intervention. By support of the Federal Ministry of Education and Research and under the auspices of the Forschungsverbund Neurotraumatology, University of Munich, a prospective system analysis was carried out on major aspects of the pre and early clinical management at a population based level in patients with traumatic brain injury. Documentation of pertinent data was made from August 1998 to July 1999 covering a catchment area of Southern Bavaria (5.6 mio inhabitants). Altogether 528 cases identified to suffer from severe head injury (GCS < or = 8 or deteriorating to that level within 48 hrs) were enrolled following admission to the hospital and establishment of the diagnosis. Further, patients dying on the scene or during transport to the hospital were also documented, particularly as to the frequency of severe head injury as underlying cause of mortality. The analysis included also cases with additional peripheral trauma (polytrauma). The efficacy of the logistics and organization of the management was studied by documentation of prognosis-relevant time intervals, as for example until arrival of the rescue squad at the scene of an accident, until intubation and administration of fluid, or upon hospital admission until establishment of the CT diagnosis and commencement of surgery or transfer to the intensive care unit, respectively. The severity of cases studied in the present analysis is evident from a mortality of far above 40% of cases admitted to the hospital, which was increased by about 20% when including prehospital mortality. The outcome data notwithstanding, the emerging results demonstrate a high efficacy of the pre- and early clinical management, as indicated by a prompt arrival of the rescue squad at the scene, a competent prehospital and early clinical management and care, indicative of a low rate of avoidable complications. It is tentatively concluded on the basis of these findings that the patient prognosis is increasingly determined by the manifestations of primary brain damage vs. the development of secondary complications. PMID- 15335112 TI - Diagnosis and screening of fragile X syndrome: experiences in Taiwan. PMID- 15335113 TI - A step-wise diagnosis of fragile X syndrome in Taiwan. AB - Fragile X syndrome (FXS), an X-linked dominant disorder, is one of the common forms of inherited mental retardation. This project aimed at identifying fragile X syndrome patients in schools by a two-step diagnosis with questionnaire and photography followed by molecular analysis. A total of 734 children with mental retardation were enrolled from kindergartens, primary schools, junior high schools, and schools for the mentally retarded. School teachers or nurses administered the questionnaires and took pictures of the faces and hands for of the patients. After viewing of the questionnaire and photos by a geneticist, 145 cases were selected for molecular study and 11 cases were identified as having full mutations in the FMRI gene. The detection rate was 1.5% (11 in 734) in all enrolled cases, and was 7.6% (11 in 145) in those who underwent molecular test. Those affected by FXS were more likely to have simian crease (p<0.001) and a head circumference larger than the 50th percentile (p=0.0295), and those who were not affected by FXS were lower in gestational age (p=0.0243). This screening method is useful for the detection of fragile X syndrome. PMID- 15335114 TI - Comparison of breath-actuated and conventional constant-flow jet nebulizers in treating acute asthmatic children. AB - To compare the therapeutic effects of two small-volume jet nebulization devices, a breath-actuated nebulizer (BAN) and a constant-flow nebulizer (CFN), this study was conducted. Seventy-two asthmatic patients, aged 5 to 15, were randomly enrolled into this study (39 to the CFN group, 33 to the BAN group). Nebulization treatment with terbutaline solution (5.0 mg /2 ml) diluted with 2 ml isotonic saline for 10 minutes was performed after measurements of baseline spirometry, SaO2, and pulse rate. Measurements of spirometry, SaO2 and pulse rate were repeated at 0, 5, 15 and 30 minutes after the end of the nebulization treatment. After treatment, for within-group comparison, all the spirometric parameters (including FEV1, PEF and FEF25-75%) and SaO2 at various time points of both groups significantly improved. The pulse rate also significantly increased (only at 15 and 30 min for the CFN group). For between-group comparison, the BAN group had greater improvement in all the data of spirometric parameters and SaO2 at various time points, but only reached statistical significance at some time points in PEF, FEF25-75% and SaO2. The pulse rate of the BAN group was significantly higher than that of the CFN group beginning 5 minutes after treatment. In conclusion, the BAN device is better than the CFN device in improving the spirometric parameters and oxygen saturation of asthmatic children during an equal 10-minute nebulization therapy. These results, associated with the higher pulse rate evoked in the BAN group, suggest a higher lung deposition ratio in the BAN device. PMID- 15335115 TI - Increased interleukin-12 release from peripheral blood mononuclear cells in nephrotic phase of minimal change nephrotic syndrome. AB - The pathogenesis of minimal change nephrotic syndrome (MCNS) is still not clear. Several studies indicate that MCNS is a systemic disorder of cell-mediated immunity. IL-12 is a pleiotropic cytokine that is produced primarily by macrophage and plays a primary role in the induction of Th1-mediated immunity. The purpose of this study is to explore the relationship of in vivo IL-12 levels in both sera and urine, and in vitro IL-12 production in stable or nephrotic clinical condition patients with MCNS. In vitro IL-12 production was detected from LPS-stimulated peripheral blood mononuclear cells (PBMNCs) of 20 MCNS patients during nephrotic and remission stages, levels of sera and urinary IL-12 were measured by commercially available ELISA kits (R & D Systems, Minneapolis). The results showed sera IL-12 levels and LPS-stimulated IL-12 productions for PBMNCs were significantly increased as compared with those of normal controls and in remission stage. However, there was also statistical difference of urinary IL 12 levels among MCNS patients during nephrotic and remission stage and controls. This study demonstrates a correlation between in vitro IL-12 release by PBMNCs and in vivo sera IL-12 level from MCNS patients as well as correlation with disease activity. The study contributes to the understanding of the pathogenesis of MCNS. PMID- 15335116 TI - Peritoneal dialysis in treatment of postoperative heart failure after cardiac surgery in infants. AB - The aim of this study was to investigate the effect of peritoneal dialysis (PD) on fluid balance and renal function in 17 infants with low cardiac output syndrome after open heart surgery. Peritoneal catheters were inserted prophylactically at the end of the cardiac operation. An effective negative fluid balance was achieved in all infants (P<0.01). They had significantly increased diuresis (P<0.01), increased mean arterial blood pressure (P<0.05), decreased central venous pressure (P<0.05), and decreased dopamine medication (P<0.05) after PD compared to before PD. Complications of PD, present in 7 patients, were transient and of minor significance. No patients died during PD or required discontinuation of PD due to severe cardiorespiratory distress. Thirteen patients recovered and had normal function upon discharge from our ward. Four died due to severe congestive heart failure. The overall mortality rate among all 17 patients was 24%. We conclude that PD is helpful in promoting fluid removal and may be used prophylactically for those infants who are at high risk for low cardiac output after cardiac surgery. PMID- 15335117 TI - Clinical spectrum of alimentary tract duplication in children. AB - Alimentary tract duplication is a rare congenital malformations. Sixteen patients admitted to our pediatric ward were enrolled in a ten-year study over the most recent decade. In this paper, we trace their history and retrospectively analyze sex, age, clinical presentations, duplication size and location, presence of ectopic tissue, complications and additional associated anomalies and prognosis. The duplications were cystic in all cases except one tubular duplication of the colon. The cases included duplication of the stomach in 3 cases (19%); ileum in 12 cases (63%); appendix in 1 case (5%); colon in 3 cases (16%); and rectum in 1 case (5%). The most frequent symptom was intermittent vomiting, half of which occurred before 1 year of age. Other symptoms included abdominal pain (50%), abdominal distension (30%), palpable mass (20%), peritoneal sign (13%), bloody stool (6%), fever (6%), and constipation (6%). The outcome was good, except in 2 patients, 1 of whom died of a perforation of the sigmoid colon duplication, and another who was complicated with multiple congenital anomalies. CONCLUSION: In our series, the average time period from the initial onset of symptoms to the diagnosis was 6.9 days, with only one exception (a 17-year-old girl who had habitual constipation). The constipation in the single exception was not a specific presentation, which may have delayed the diagnosis. Pediatric patients with unexplained gastrointestinal symptoms must be diagnosed for the possibility of alimentary tract duplication, even though it rarely occurs. Ultrasonography is helpful in diagnosing a duplication, as it will identify a three-layered image representing the duplication cyst, common wall, and outer bowel wall. These features have proven useful for establishing the correct diagnosis or ruling out alimentary tract duplication where surgery was indicated anyway. PMID- 15335118 TI - Hyposensitization therapy reduced exhaled nitric oxide in asthmatic children with corticosteroid dependency. AB - Measurement of exhaled nitric oxide (FeNO) is a simple and noninvasive method for assessment of airway inflammation severity in asthmatic patients. The aim of this study was to investigate whether hyposensitization with extracts of Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f) could decrease airway inflammation as determined by FeNO levels. Ten asthmatic children with corticosteroid dependency were recruited for the study. Eight subjects completed a ten months mite hyposensitization regimen. FeNO concentrations, asthma symptom scores and peak expiratory flow rates (PEFR) were measured monthly before and after treatment. FeNO levels from asthmatic patients were found to be higher before treatment (65.34+/-31.63 ppb). FeNO levels fell significantly after four months' treatment (39.89+/-21.13 ppb; p<0.05). PEFR increased significantly five months after hyposensitization treatment (214.38+/-49.38 vs 251.25+/-28.00; p<0.05). Asthma symptom scores decreased 5 months after hyposensitization treatment (6.84+/-1.22 vs 5.80+/-0.67, p<0.05). Hyposensitization with extracts of Der p and Der f appears to improve clinical symptoms and reduce airway inflammation as determined by FeNO concentrations in children with mite allergy asthma. PMID- 15335119 TI - Kawasaki disease associated with chickenpox: report of two sibling cases. AB - Kawasaki disease is a group of acute febrile illnesses of young children. Infectious etiology for Kawasaki disease has been long postulated, although no single causing agent has been identified consistently by conventional methods. This report described two sisters, soon after a primary infection by varicella zoster virus, manifesting fever and muco-cutaneous lesions compatible with the feature of Kawasaki disease consecutively. Mild dilatation of the coronary artery was revealed in both by echocardiography. Both of them received intravenous gamma globulin therapy and the follow-up echocardiography showed no further progression of the coronary lesion. We suggest the possibility of varicella-zoster virus as one of the triggering agents for Kawasaki disease in genetically susceptible individuals. The severe cardiac complication of Kawasaki disease may be prevented by early treatment if clinicians are alert to this possible association in those children with chickenpox exhibiting unusual muco-cutaneous lesions. PMID- 15335120 TI - Partial trisomy 1 with congenital hydrocephalus and hypogammaglobulinemia: report of one case. AB - We report a male patient with a 46, XY, der (11) t (1; 11) (q31; q25) karyotype due to de novo unbalanced translocation. The boy had facial dysmorphism including prominent wide forehead, short bilateral palpebral fissures, broad nasal bridge, low set and malformed ears, digitalization of thumbs, and small testes. Besides, he also suffered from congenital hydrocephalus and hypogammaglobulinemia, which have not been described in trisomy 1q syndrome. The additional chromatin material on the long arm of chromosome 11 segment was derived from chromosome 1, as proved by high-resolution banding and multiple-color FISH study. This case report allows a further delineation of the trisomy 1q syndrome. PMID- 15335121 TI - Cellulitis resulting from infection by Haemophilus influenzae type b: report of two cases. AB - Cellulitis resulting from infection by Haemophilus influenzae type b (Hib) is uncommon but may be a characteristic of such infection. Especially in the post Hib vaccine era, such infection represents a potentially life-threatening infectious process due to the relatively high incidence of accompanying bacteremia and meningitis. Here, we report on two apparently previously healthy children who developed Hib cellulitis, one of whom revealed lower-leg cellulitis while the other exhibited buccal cellulitis associated with an oral ulcer. To the best of our knowledge, there have been no formal reports pertaining to buccal cellulitis following infection by Hib yet published in Taiwan, and reports of Hib cellulitis of the extremities still appear to be rare, hence we report on these two cases. PMID- 15335122 TI - Moyamoya disease initially mimicking MELAS syndrome in a 14-month-old child. AB - Moyamoya disease is a neurological disease rarely seen in children outside Japan. It is difficult to differentiate moyamoya disease in its early stage from mitochondrial disorders in children when nontraumatic ischemic stroke is considered. We present a 14-month-old Taiwanese female child who had suffered from sudden onset of nontraumatic seizure attack, after which progressive left limb weakness was noted. Initial brain imaging and a series of laboratory studies were performed which suggested mitochondrial disorders, especially mitochondrial encephalopathy, lactic acidosis, and strokelike episodes (MELAS) syndrome. The patient underwent an oral glucose lactate stimulation test (OGLST), and mitochondrial disorders were not favored. MR angiography of the circle of Willis showed moyamoya disease. The patient then underwent an encephalo-duro-arterio-myo synangiosis (EDAMS) operation. Evaluation of the neurodevelopmental and intelligence outcome required close long-term follow-up. PMID- 15335123 TI - Early diagnosis of Prader-Willi syndrome in a newborn. AB - Prader-Willi syndrome (PWS) is a multiple-systemic disorder with many manifestations related to hypothalamic insufficiency, with obesity and behavioral problems as the major causes of morbidity and mortality. We describe a 2-day-old boy who initially presented with neonatal hypotonia and was diagnosed as PWS based on abnormal DNA methylation patterns in the small nuclear ribonucleoprotein polypeptide N (SNRPN) gene at the age of one week, despite the absence of other classical features. Molecular diagnosis for PWS, which has become available in recent years, should be considered for neonates with undiagnosed central hypotonia. PMID- 15335124 TI - Recurrent peritonitis with massive ascites as the initial manifestation of systemic lupus erythematosus: report of one case. AB - Systemic lupus erythematosus (SLE) rarely presents with massive ascites secondary to severe peritonitis before the occurrence of major diagnostic features. We described a 13-year-old girl who developed four episodes of severe abdominal pain with massive ascites. During the first episode, the patient experienced deterioration with massive ascites over a 3-day period, after a 3-month history of intermittent abdominal pain. This episode resolved under conservative treatment. During the second episode, her acute abdominal findings mimicked appendicitis or gynecologic emergency. An exploratory laparotomy was initially planned, but not performed. Thrombocytopenia postponed the surgical intervention and led to further investigation. Autoimmune studies confirmed SLE and prompted steroid therapy, which rapidly improved her painful ascites and spared an unnecessary operation. After improvement of peritonitis with ascites, and following discharge, other major clinical features of SLE ensued. The third episode of peritonitis with massive ascites responded well to steroid pulse therapy. The fourth episode of ascites was accompanied by intracranial hemorrhage that led to her death. To the best of our knowledge, this is the first case report of recurrent peritonitis with massive ascites as the initial and major manifestation of SLE in a pediatric patient. PMID- 15335125 TI - Why isn't it better? PMID- 15335126 TI - A randomized controlled trial to increase cancer screening among attendees of community health centers. AB - BACKGROUND: We assessed the efficacy of the Cancer Screening Office Systems (Cancer SOS), an intervention designed to increase cancer screening in primary care settings serving disadvantaged populations. METHODS: Eight primary care clinics participating in a county-funded health insurance plan in Hillsborough County, Fla, agreed to take part in a cluster-randomized experimental trial. The Cancer SOS had 2 components: a cancer-screening checklist with chart stickers that indicated whether specific cancer-screening tests were due, ordered, or completed; and a division of office responsibilities to achieve high screening rates. Established patients were eligible if they were between the ages of 50 and 75 years and had no contraindication for screening. Data abstracted from charts of independent samples collected at baseline (n = 1,196) and at a 12-month follow up (n = 1,237) was used to assess whether the patient was up-to-date on one or more of the following cancer-screening tests: mammogram, Papanicolaou (Pap) smear, or fecal occult blood testing (FOBT). RESULTS: In multivariate analysis that controlled for baseline screening rates, secular trends, and other patient and clinic characteristics, the intervention increased the odds of mammograms (odds ratio [OR] = 1.62, 95% confidence interval [CI], 1.07-9.78, P = .023) and fecal occult blood tests (OR = 2.5, 95% CI, 1.65-4.0, P <.0001) with a trend toward greater use of Pap smears (OR = 1.57, 95% CI, 0.92-2.64, P = .096). CONCLUSIONS: The Cancer SOS intervention significantly increased rates of cancer screening among primary care clinics serving disadvantaged populations. The Cancer SOS intervention is one option for providers or policy makers who wish to address cancer related health disparities. PMID- 15335127 TI - Going to scale: re-engineering systems for primary care treatment of depression. AB - BACKGROUND: Recent trials have shown improved depression outcomes with chronic care models. We report the methods of a project that assesses the sustainability and transportability of a chronic care model for depression and change strategy. METHODS: In a randomized controlled trial (RCT), a clinical model for depression was implemented through a strategy supporting practice change. The clinical model is evidence based. The change strategy relies on established quality improvement programs and is informed by diffusion of innovations theory. Evaluation will address patient outcomes, as well as process of care and process of change. RESULTS: Five medical groups and health plans are participating in the trial. The RCT involves 180 clinicians in 60 practices. All practices assigned to the clinical model have implemented it. Participating organizations have the potential to disseminate this clinical model of care to 700 practices and 1,700 clinicians. CONCLUSIONS: It is feasible to implement the clinical model and change strategy in diverse practices. Follow-up evaluation will determine the impact, sustainability, and potential for dissemination. Materials are available through http://www. depression-primarycare.org; more in-depth descriptions of the clinical model and change strategy are available in the online-only appendixes to this article. PMID- 15335128 TI - Primary care clinicians evaluate integrated and referral models of behavioral health care for older adults: results from a multisite effectiveness trial (PRISM e). AB - BACKGROUND: Recent studies have shown that integrated behavioral health services for older adults in primary care improves health outcomes. No study, however, has asked the opinions of clinicians whose patients actually experienced integrated rather than enhanced referral care for depression and other conditions. METHOD: The Primary Care Research in Substance Abuse and Mental Health for the Elderly (PRISM-E) study was a randomized trial comparing integrated behavioral health care with enhanced referral care in primary care settings across the United States. Primary care clinicians at each participating site were asked whether integrated or enhanced referral care was preferred across a variety of components of care. Managers also completed questionnaires related to the process of care at each site. RESULTS: Almost all primary care clinicians (n = 127) stated that integrated care led to better communication between primary care clinicians and mental health specialists (93%), less stigma for patients (93%), and better coordination of mental and physical care (92%). Fewer thought that integrated care led to better management of depression (64%), anxiety (76%), or alcohol problems (66%). At sites in which the clinicians were rated as participating in mental health care, integrated care was highly rated as improving communication between specialists in mental health and primary care. CONCLUSIONS: Among primary care clinicians who cared for patients that received integrated care or enhanced referral care, integrated care was preferred for many aspects of mental health care. PMID- 15335129 TI - Preventing errors in clinical practice: a call for self-awareness. AB - While ascribing medical errors primarily to systems factors can free clinicians from individual blame, there are elements of medical errors that can and should be attributed to individual factors. These factors are related less commonly to lack of knowledge and skill than to the inability to apply the clinician's abilities to situations under certain circumstances. In concert with efforts to improve health care systems, refining physicians' emotional and cognitive capacities might also prevent many errors. In general, physicians have the sensation of making a mistake because of the interference of emotional elements. We propose a so-called rational-emotive model that emphasizes 2 factors in error causation: (1) difficulty in reframing the first hypothesis that goes to the physician's mind in an automatic way, and (2) premature closure of the clinical act to avoid confronting inconsistencies, low-level decision rules, and emotions. We propose a teaching strategy based on developing the physician's insight and self-awareness to detect the inappropriate use of low-level decision rules, as well as detecting the factors that limit a physician's capacity to tolerate the tension of uncertainty and ambiguity. Emotional self-awareness and self regulation of attention can be consciously cultivated as habits to help physicians function better in clinical situations. PMID- 15335130 TI - A string of mistakes: the importance of cascade analysis in describing, counting, and preventing medical errors. AB - BACKGROUND: Notions about the most common errors in medicine currently rest on conjecture and weak epidemiologic evidence. We sought to determine whether cascade analysis is of value in clarifying the epidemiology and causes of errors and whether physician reports are sensitive to the impact of errors on patients. METHODS: Eighteen US family physicians participating in a 6-country international study filed 75 anonymous error reports. The narratives were examined to identify the chain of events and the predominant proximal errors. We tabulated the consequences to patients, both reported by physicians and inferred by investigators. RESULTS: A chain of errors was documented in 77% of incidents. Although 83% of the errors that ultimately occurred were mistakes in treatment or diagnosis, 2 of 3 were set in motion by errors in communication. Fully 80% of the errors that initiated cascades involved informational or personal miscommunication. Examples of informational miscommunication included communication breakdowns among colleagues and with patients (44%), misinformation in the medical record (21%), mishandling of patients' requests and messages (18%), inaccessible medical records (12%), and inadequate reminder systems (5%). When asked whether the patient was harmed, physicians answered affirmatively in 43% of cases in which their narratives described harms. Psychological and emotional effects accounted for 17% of physician-reported consequences but 69% of investigator-inferred consequences. CONCLUSIONS: Cascade analysis of physicians' error reports is helpful in understanding the precipitant chain of events, but physicians provide incomplete information about how patients are affected. Miscommunication appears to play an important role in propagating diagnostic and treatment mistakes. PMID- 15335131 TI - Event reporting to a primary care patient safety reporting system: a report from the ASIPS collaborative. AB - BACKGROUND: We examined reports to a primary care, ambulatory, patient safety reporting system to describe types of errors reported and differences between anonymous and confidential reports. METHODS: Applied Strategies for Improving Patient Safety (ASIPS) is a demonstration project designed to collect and analyze medical error reports from clinicians and staff in 2 practice-based research networks: the Colorado Research Network (CaReNet) and the High Plains Research Network (HPRN). A major component of ASIPS is a voluntary patient safety reporting system that accepts reports of errors anonymously or confidentially. Reports are coded using a multiaxial taxonomy. RESULTS: Two years into this project, 33 practices with a total of 475 clinicians and staff have participated in ASIPS. Participants submitted 708 reports during this time (66% using the confidential reporting form). We successfully followed up on 84% of the confidential reports of interest within the allotted 10-day time frame. We ended up with 608 relevant, codable reports. Communication problems (70.8%), diagnostic tests (47%), medication problems (35.4%), and both diagnostic tests and medications (13.6%) were the most frequently reported errors. Confidential reports were significantly more likely than anonymous reports to contain codable data. CONCLUSION: A safe and secure reporting system that relies on voluntary reporting from clinicians and staff can be successfully implemented in primary care settings. Information from confidential reports appears to be superior to that from anonymous reports and may be more useful in understanding errors and designing interventions to improve patient safety. PMID- 15335132 TI - Patient reports of preventable problems and harms in primary health care. AB - BACKGROUND: Despite recent attention given to medical errors, little is known about the kinds and importance of medical errors in primary care. The principal aims of this study were to develop patient-focused typologies of medical errors and harms in primary care settings and to discern which medical errors and harms seem to be the most important. METHODS: Thirty-eight in-depth anonymous interviews of adults from rural, suburban, and urban locales in Virginia and Ohio were conducted to solicit stories of preventable problems with primary health care that led to physical or psychological harm. Transcriptions were analyzed to identify, name, and organize the stories of errors and harms. RESULTS: The 38 narratives described 221 problematic incidents that predominantly involved breakdowns in the clinician-patient relationship (n = 82, 37%) and access to clinicians (n = 63, 29%). There were several reports of perceived racism. The incidents were linked to 170 reported harms, 70% of which were psychological, including anger, frustration, belittlement, and loss of relationship and trust in one's clinician. Physical harms accounted for 23% of the total and included pain, bruising, worsening medical condition, and adverse drug reactions. DISCUSSION: The errors reported by interviewed patients suggest that breakdowns in access to and relationships with clinicians may be more prominent medical errors than are technical errors in diagnosis and treatment. Patients were more likely to report being harmed psychologically and emotionally, suggesting that the current preoccupation of the patient safety movement with adverse drug events and surgical mishaps could overlook other patient priorities. PMID- 15335133 TI - Appropriateness of Lyme disease serologic testing. AB - BACKGROUND: Although rapid diagnosis of Lyme disease is essential for effective treatment, there is concern about inappropriate testing. We conducted a prospective, cross-sectional survey of clinicians to assess the use and appropriateness of Lyme disease serologic tests (LDSTs). METHODS: LDSTs performed at 2 large Wisconsin reference laboratories were systematically sampled for 12 consecutive months. A standardized questionnaire was used to gather data about the submitting clinician and the patient tested. Tests were categorized as appropriate, inappropriate, or discretionary, and associations were assessed using logistic regression analysis. A test was defined as inappropriate if the patient was asymptomatic, had erythema migrans, or was treated empirically, or if the test was ordered as a test of cure. RESULTS: We surveyed 303 clinicians regarding 356 LDSTs: 72 tests (20%) were appropriate, 95 (27%) were inappropriate, and 189 (53%) were discretionary. Tests were more likely to be inappropriate if they were ordered by an emergency or urgent care physician compared with other specialists (adjusted odds ratio [AOR] 5.2, 95% confidence interval [CI], 1.3-20.6), or if preceded by a known tick bite (AOR 6.8, 95% CI, 2.6-17.6). The patient rather than the clinician requested 26% of tests, which were more likely to be inappropriate than clinician-requested tests (crude odds ratio [COR] 5.8, 95% CI, 2.5-13.6). Tests were more likely to be patient requested if they were ordered by an internist (AOR 2.6, 95% CI, 1.4-4.8) or if the patient was > or =40 years old (AOR 2.2, 95% CI, 1.3-3.9). CONCLUSIONS: Many LDSTs are ordered inappropriately, often influenced by patient demand. Education of clinicians and patients about testing indications and contraindications is needed to reduce the number of inappropriate LDSTs. PMID- 15335134 TI - The visit before the morning after: barriers to preprescribing emergency contraception. AB - BACKGROUND: Research suggests that while advance prescription of emergency contraception (EC) increases women's access, this prescribing model is rarely used. The present study sought to explore attitudes towards EC among patients and physicians, with the goal of understanding potential barriers to advance prescription. METHODS: Qualitative, semistructured interviews were conducted with patients and clinicians in a New York City family practice clinic. RESULTS: Using qualitative interviews, we found that attitudes towards EC among patients and clinicians are complex. Both groups of participants reported favorable attitudes towards EC. There was general agreement that physicians should take a proactive role in educating patients about the method. A notable minority in each group described substantial reservations, however, especially regarding the potential for EC abuse. Such attitudes emerged mainly in the context of discussions about advance prescription. Advance prescription was viewed as greatly facilitating access to EC, but some patients and clinicians feared that ready access would encourage irresponsible sex. Some participants condoned the occasional, accidental, or emergency use of EC; however, habitual use, or the plan not to plan for sex, was viewed as morally indefensible. CONCLUSION: Findings suggest that even when attitudes towards EC are generally favorable, some physicians and patients have substantial reservations about advance prescription. Education and dialogue are needed to overcome these reservations. PMID- 15335135 TI - Subclinical hypothyroidism and the risk of hypercholesterolemia. AB - BACKGROUND: Subclinical hypothyroidism, defined as a mild elevation in thyroid stimulating hormone (TSH) levels in patients with normal serum thyroxine levels, has been associated with elevationed levels in serum cholesterol in some sample populations. These studies, however, have included referred patients and large numbers of patients with previously treated hyperthyroidism. The aim of this study was to assess whether subclinical hypothyroidism is associated with abnormal lipid levels in a population-based sample. METHODS: Data from adults older than 40 years who did not previously have a diagnosis of hypothyroidism or who were taking thyroid replacement medication were analyzed from the National Health and Nutritional Examination Survey (NHANES) III. Subclinical hypothyroidism was defined as a TSH value of 6.7 to 14.9 mU/L and normal thyroxine (n = 215). Euthyroid control adults included participants with a TSH in a normal range between 0.36 and 6.7 mU/L (n = 8,013). Outcomes examined were serum cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglyceride levels in those who had subclinical hypothyroidism and in euthyroid controls. RESULTS: Persons meeting the criteria for subclinical hypothyroidism had higher mean cholesterol levels (226 vs 217 mg/dL, P = .003) and rates of elevated cholesterol levels (74.2% vs 63.9%, P = 0.02) than the euthyroid control group, but there were no significant differences in low-density lipoprotein (LDL) or high-density lipoprotein (HDL) levels. When adjusted for age, race, sex, and the use of lipid-lowering drugs, however, subclinical hypothyroidism was not related to elevations in cholesterol levels (adjusted odds ratio [OR] = 1.06, 95% confidence interval [CI], 0.57-1.97), LDL levels (adjusted OR = 0.89; 95% CI, 0.59-1.35), or triglyceride levels (adjusted OR = 1.83; 95% CI, 0.87-3.85) or to a low HDL level (adjusted OR = 0.94; 95% CI, 0.36-2.48). CONCLUSIONS: Subclinical hypothyroidism does not appear to be associated with abnormalities in serum cholesterol or triglyceride levels when adjusted for confounding variables in this population-based study. PMID- 15335136 TI - Discussing spirituality with patients: a rational and ethical approach. AB - BACKGROUND: This study was undertaken to determine when patients feel that physician inquiry about spirituality or religious beliefs is appropriate, reasons why they want their physicians to know about their spiritual beliefs, and what they want physicians to do with this information. METHODS: Trained research assistants administered a questionnaire to a convenience sample of consenting patients and accompanying adults in the waiting rooms of 4 family practice residency training sites and 1 private group practice in northeastern Ohio. Demographic information, the SF-12 Health Survey, and participant ratings of appropriate situations, reasons, and expectations for physician discussions of spirituality or religious beliefs were obtained. RESULTS: Of 1,413 adults who were asked to respond, 921 completed questionnaires, and 492 refused (response rate = 65%). Eighty-three percent of respondents wanted physicians to ask about spiritual beliefs in at least some circumstances. The most acceptable scenarios for spiritual discussion were life-threatening illnesses (77%), serious medical conditions (74%) and loss of loved ones (70%). Among those who wanted to discuss spirituality, the most important reason for discussion was desire for physician patient understanding (87%). Patients believed that information concerning their spiritual beliefs would affect physicians' ability to encourage realistic hope (67%), give medical advice (66%), and change medical treatment (62%). CONCLUSIONS: This study helps clarify the nature of patient preferences for spiritual discussion with physicians. PMID- 15335137 TI - Screening for syphilis infection: recommendation statement. PMID- 15335138 TI - Pounds. AB - This is the story of a loving relationship between a doctor and a vibrant, elderly woman. In the story, the doctor learns to accept his patient's noncompliance with medical regimens; at the same time, he learns something about his own noncompliance. PMID- 15335139 TI - Stephen Zyznski receives 2004 Curtis G. Hames Research Award. PMID- 15335140 TI - A different perspective. PMID- 15335141 TI - Pilot project studies EHR implementation issues. PMID- 15335142 TI - The effective lymphatic absorption rate is an accurate and useful concept in the physiology of peritoneal dialysis. PMID- 15335143 TI - Effective lymphatic absorption rate is not a useful or accurate term to use in the physiology of peritoneal dialysis. PMID- 15335144 TI - Arterial stiffness and renal replacement therapy. PMID- 15335145 TI - Now you see them, now you don't. PMID- 15335146 TI - Chronic inflammation in peritoneal dialysis: the search for the holy grail? AB - Mortality and morbidity in chronic kidney disease (CKD) patients are unacceptably high. The annual mortality rate due to cardiovascular disease (CVD) is approximately 9%, which, for the middle-aged person, is at least 10- to 20-fold higher than for the general population. Classic risk factors for CVD are highly prevalent in CKD patients, but they cannot fully account for the excessive rate of CVD in this population. Instead, it has become increasingly clear that nontraditional risk factors, such as systemic inflammation, may play a key role in the development of atherosclerosis. It is well established that inflammatory markers are very powerful predictors of high CVD morbidity and mortality not only in the general population, but particularly in CKD patients. Signs of a sustained low-grade inflammation, such as increased levels of C-reactive protein (CRP), are present in the majority of stage 5 CKD patients, even in patients in clinically stable condition, and they are also commonly observed after the initiation of dialysis therapy. Dialysis therapy--hemodialysis as well as peritoneal dialysis (PD)--may itself contribute to systemic inflammation. Local intraperitoneal inflammation can also occur in patients treated with PD. These local effects may result in a low-grade inflammation, caused by the bioincompatibility of conventional glucose-based dialysis fluids, to intense inflammation associated with peritonitis. Given these circumstances, it is reasonable to hypothesize that strategies aiming to reduce inflammation are potentially important and novel, and could serve to reduce CVD, thereby lowering morbidity and mortality in patients with CKD. In this review we provide information supporting the hypothesis that systemic inflammation is tightly linked to the most common complications of CKD patients, in particular those on PD, and that local inflammation in PD may contribute to various related complications. The aims of this review are to discuss the reasons that make inflammation an attractive target for intervention in CKD, the particular aspects of the inflammation-CVD axis during PD treatment that are likely involved, and possible means for the detection and management of chronic inflammation in PD patients. PMID- 15335147 TI - Obesity is a risk factor for peritonitis in the Australian and New Zealand peritoneal dialysis patient populations. AB - OBJECTIVE: The aim of the present investigation was to examine the association between body mass index (BMI) and peritonitis rates among incident peritoneal dialysis (PD) patients in a large cohort with long-term follow-up. DESIGN: Retrospective observational cohort study of the Australian and New Zealand PD patient population. SETTING: Australia and New Zealand Dialysis and Transplant (ANZDATA) Registry. PARTICIPANTS: The study included all incident adult patients (n = 10 709) who received PD in Australia and New Zealand in the 12-year period between 1 April 1991 and 31 March 2003. Patients were classified as obese (BMI > or = 30 kg/m2), overweight (BMI 25.0 - 29.9 kg/m2), normal weight (20 - 24.9 kg/m2), or underweight (< 20 kg/m2). MAIN MEASUREMENTS: Time to first peritonitis and episodes of peritonitis per patient-year were recorded over the 12-year period. RESULTS: Higher BMI was associated with a shorter time to first peritonitis episode, independent of other risk factors [hazard ratio 1.08 for each 5-kg/m2 increase in BMI, 95% confidence interval (CI) 1.04 - 1.12, p < 0.001]. When peritonitis outcomes were analyzed as episodes of peritonitis per patient-year, these rates were significantly higher among patients with higher BMI: underweight 0.69 episodes/year (95% CI 0.66 - 0.73), normal weight 0.79 (95% CI 0.77 - 0.81), overweight 0.88 (95% CI 0.85 - 0.90), obese 1.06 (95% CI 1.02 - 1.09). Coronary artery disease and chronic lung disease were associated with both shorter time to first peritonitis and higher peritonitis rates, independently of these other factors. There was also a "vintage effect," with lower peritonitis rates seen among people who commenced dialysis in more recent years. CONCLUSIONS: Higher BMI at the commencement of renal replacement therapy is a significant risk factor for peritonitis. The mechanisms for this remain undefined. PMID- 15335148 TI - Does lymphatic absorption change with the duration of peritoneal dialysis? AB - BACKGROUND: Ultrafiltration failure is an important complication of long-term peritoneal dialysis (PD). A high effective lymphatic absorption rate (ELAR) can contribute to impaired ultrafiltration. It is unknown whether the ELAR increases with time on PD. OBJECTIVE: The relationship between the ELAR and duration of PD was analyzed, as well as the correlation between the ELAR and other transport parameters. We also studied the relation between the ELAR and cancer antigen 125 (CA125) a marker for mesothelial cell mass. SETTING: Peritoneal dialysis unit in the Academic Medical Center, Amsterdam. DESIGN: Cross-sectional and longitudinal studies of standard peritoneal permeability analyses (SPAs; 4-hour dwells, dextran 70 as a volume marker) with glucose 3.86% in 130 PD patients. METHODS: SPAs were analyzed in 130 stable PD patients (77 males). Median duration of PD was 25 months (range 1-214) in a cross-sectional study. The last SPA from each patient was analyzed. The longitudinal analysis included 24 patients (12 males) from whom at least 3 SPAs were available with a minimum interval of 8 months. Dextran 70, 1 g/L, was administered intraperitoneally at the initiation of the test. Lymphatic absorption was calculated from the disappearance rate of dextran 70 during the 4-hour dwell. Therefore, the ELAR included both transmesothelial and subdiaphragmatic uptake of dextran 70. RESULTS: Median ELAR was 1.43 mL/minute (range 0.17- 6.59 mL/minute). No relationship was found between the ELAR and duration of PD in the cross-sectional analysis, nor was there a trend in time for 20 of the 24 patients studied longitudinally. In 4 patients, a negative trend was found. None of these had ultrafiltration failure and all 4 patients had a different cause for end-stage renal failure. The ELAR was correlated with parameters of peritoneal solute transport, but not with CA125 when investigated in a cross-sectional analysis. Only after 48 months of PD treatment was a significant relationship between the ELAR and CA125 seen (r = 0.46, p < 0.05). CONCLUSIONS: No time trend is present for the effective peritoneal lymphatic absorption rate, and it is not associated with patient or technique survival. Although increased lymphatic absorption is one of the causes of ultrafiltration failure, it is unlikely to contribute to the development of ultrafiltration failure in long-term PD patients with well-maintained transcapillary ultrafiltration. PMID- 15335149 TI - The ratio of extracellular fluid to total body water and technique survival in peritoneal dialysis patients. AB - BACKGROUND: Patients receiving peritoneal dialysis experience a high technique failure rate and are often overhydrated. We examined whether an increased extracellular fluid volume (VECF) as a proportion of the total body water (VTBW) predicted technique survival (TS) in a prevalent patient cohort. METHODS: The VECF and VTBW were estimated by multiple-frequency bioelectric impedance in 59 prevalent peritoneal dialysis patients (median time on dialysis 14 months). Demographic, biochemical (albumin, C-reactive protein, and ferritin), and anthropometric data, forearm muscle strength, nutritional score by three-point Subjective Global Assessment, residual renal function, dialysate-to-plasma (D/P) creatinine ratio, total weekly Kt/V urea, total creatinine clearance, normalized protein equivalent of nitrogen appearance, and midarm muscle circumference were also assessed. Technique survival was determined at 3 years, and significant predictors of TS were sought. RESULTS: In patient groups defined by falling above or below the median value for each parameter, only residual renal function (p = 0.002), 24-hour ultrafiltrate volume (p = 0.02), and VECF/VTBW ratio (p = 0.05) were significant predictors of TS. Subjects with a higher than median VECF/VTBW ratio had a 3-year TS of 46%, compared to 78% in subjects with a lower than median value. In multivariate analysis, systolic blood pressure and VECF/VTBW ratio (both p < 0.05) were significant predictors of TS. C-reactive protein approached significance. CONCLUSION: Increased ratio of extracellular fluid volume to total body water is associated with decreased TS in peritoneal dialysis. PMID- 15335150 TI - The self-locating catheter: clinical experience and follow-up. AB - BACKGROUND: The self-locating catheter invented by Nicola Di Paolo has been used increasingly in Italy and elsewhere since 1994, with about a thousand patients currently implanted every year. Twelve grams of tungsten inserted into the tip of the conventional Tenckhoff catheter during extrusion does not significantly change its form, but suffices to keep the tip firmly in the Douglas cavity. OBJECTIVE: The aim of the present study was to confirm our preliminary results in a large population of peritoneal dialysis patients. SETTING: 16 Italian nephrology departments. RESULTS: In addition to confirming the validity of the new catheter, the present results show that patients with the new catheter have fewer episodes of peritonitis, tunnel infection, cuff extrusion, catheter malfunction, obstruction, and leakage. CONCLUSION: The present multicenter control study confirms preliminary results and demonstrates that complications of peritoneal dialysis, such as cuff extrusion, infection, peritonitis, early leakage, and obstruction, are statistically less frequent in patients with self locating catheters than in patients with classic Tenckhoff catheters. PMID- 15335151 TI - The influence of dialytic modality on arterial stiffness, pulse wave reflections, and vasomotor function. AB - BACKGROUND: Measurements of aortic stiffness [aortic pulse wave velocity (PWV) and augmentation index (Alx)] have been established as powerful predictors of survival on hemodialysis (HD). Abnormal endothelial-dependent and endothelial independent vascular reactivity and increased arterial stiffness are commonly described in HD patients. There is, however, a lack of information on the comparative impact of different renal replacement therapies (RRTs) on PWV and Alx, and how these different methods might influence endothelial-dependent abnormal vasodilatation. OBJECTIVE: To describe in a cross-sectional design arterial compliance and distensibility in continuous ambulatory peritoneal dialysis (CAPD) versus HD versus renal transplant (RTx) patients, compared with age- and blood pressure-matched essential hypertensive controls. The PWV and aortic Alx were determined from contour analysis of arterial waveforms recorded by applanation tonometry in 40 CAPD, 41 HD, 20 RTx patients (with normal serum creatinine), and 20 controls with essential hypertension (all normotensive under treatment). Endothelial-dependent and endothelial-independent vascular reactivities were assessed by changes in Alx following challenges with inhaled salbutamol and sublingual nitroglycerin respectively. RESULTS: CAPD patients had significantly stiffer arteries than all other categories. The PWV was 8.29 +/- 1.09 m/ second in CAPD patients, significantly higher (p < 0.05) compared to HD subjects (7.19 +/- 1.87 m/s). Both dialysis subgroups had significantly higher PWV values compared to RTx patients (6.59 +/- 1.62 m/s) and essential hypertensive controls (6.34 +/- 1.32 m/s), p < 0.05. The Alx had a profile similar to PWV in different RRTs. All groups with the exception of CAPD subjects had a significant decrease in Alx following salbutamol. Moreover, the vasodilatation induced by either nitroglycerin or salbutamol was significantly blunted compared to HD. Overall, both dialysis categories had more abnormal responses compared to RTx patients and essential hypertensive controls. CONCLUSION: CAPD is associated with stiffer arteries and more profoundly abnormal endothelial-dependent vasomotor function, compared to matched HD subjects. These differences in arterial physical properties might explain differences seen in cardiac structure and function between the RRTs. PMID- 15335152 TI - Effective homocysteine-lowering vitamin B treatment in peritoneal dialysis patients. AB - BACKGROUND: Hyperhomocysteinemia, a risk factor for atherosclerosis, is frequently detected in patients with renal failure. Vitamin B supplementation reduces but rarely normalizes homocysteine (Hcy) levels in hemodialysis patients. There are no data about the effects of vitamin B therapy on Hcy levels in patients on peritoneal dialysis (PD). AIMS: We performed this trial both to observe baseline plasma Hcy levels in PD patients and to assess the effects of vitamin B therapy on Hcy levels in continuous ambulatory PD patients. METHODS: We conducted a 6-month prospective study of the effects of vitamin B therapy on plasma Hcy levels. Biochemical analyses were obtained at baseline and after every phase of treatment with folic acid, folic acid plus vitamin B12, and folic acid plus vitamin B12 plus vitamin B6. Eighteen of the 25 enrolled patients finished the study. RESULTS: Hyperhomocysteinemia was present in 83% of PD patients. We detected a trend toward a significant inverse relationship between baseline Hcy and folate levels. There were no significant correlations between baseline Hcy and vitamin B12, peritoneal membrane permeability, dialytic efficiency, or computed peritoneal Hcy clearance. We obtained an effective decrease in mean Hcy concentration from 20 to 14.8 micromol/L after folic acid and vitamin B12 treatment. We observed a further reduction in mean Hcy level to 12.8 micromol/L using the triple therapy; 72% of patients normalized their Hcy value. CONCLUSIONS: High doses of folic acid, vitamin B6, and vitamin B12 normalize Hcy values in the majority of PD patients. This treatment may be important in reducing cardiovascular morbidity and mortality. PMID- 15335153 TI - Amino acid-based peritoneal dialysis solution stimulates mesothelial nitric oxide production. AB - OBJECTIVE: Ultrafiltration failure is a common problem in continuous ambulatory peritoneal dialysis. Recent work has indicated a role of enhanced expression of nitric oxide synthase (NOS) in ultrafiltration failure. However, the conditions predisposing to increased generation of NO by the peritoneum have not been studied in detail and the cell types potentially involved have not been tested individually. DESIGN: We performed experiments in human peritoneal mesothelial cells (HPMC) in culture. Amino acid-based dialysis solution (Nutrineal; Baxter Deutschland GmbH, Munchen, Germany), L-arginine, and glucose-containing control solutions were used and we observed the effects on the HPMC. We reasoned that amino acid-based dialysis solutions containing L-arginine, the substrate of NOS, might influence mesothelial NO generation. Nitric oxide production was measured in the supernatant using the Griess reaction. We studied the effect of the combined NOS inhibitor L-NMMA and specified the isoform of NOS involved. RESULTS: In serum-free control medium, the cells exhibited baseline generation of nitrite at a rate of 5.4 +/- 0.5 micromol/g protein. Addition of 6 mmol/L L-arginine to the control medium increased nitrite significantly (11.8 +/- 0.66 micromol/g protein, p < 0.002), as did amino acid-based dialysis solution (15.7 +/- 1.3 micromol/g protein, p < 0.002); L-NMMA caused a significant reduction of this nitrite. HPMC expressed eNOS (NOSIII) when grown in L-arginine-supplemented medium, shown by immunocytochemistry and by reverse transcriptase-polymer chain reaction. Biochemical exposure to a calcium ionophore in 1 micromol/L concentration approximately doubled the nitrite production by L-arginine incubated cells. CONCLUSION: Peritoneal mesothelial cells generate NO in vitro. Generation of NO increased further in response to L-arginine supplementation of the culture medium and to amino acid-containing dialysis solution. Mesothelial cells express eNOS, which was likely involved in the observed peritoneal NO generation. PMID- 15335154 TI - Temperature: the single most important factor for degradation of glucose fluids during storage. AB - OBJECTIVE: Bioincompatible glucose degradation products (GDPs) develop during heat sterilization of peritoneal dialysis (PD) fluids. However, degradation may also take place during storage. Consequently, storage may add to the bioincompatibility caused by heat sterilization. The aim of the present study was to investigate how different factors such as the sterilization procedure, pH, glucose concentration, and temperature influence GDP production during storage. DESIGN: Degradation in glucose solutions was followed by pH and UV absorbance at 228 nm and 284 nm over 2 years of storage. Different sterilization times, storage temperatures, pH, and glucose concentrations were included in the study. Peritoneal dialysis fluids were also used in the experiment. Bioincompatibility was estimated through inhibition of cell growth in L-929 fibroblasts, and GDPs through UV absorption and liquid chromatography. RESULTS: The most important factor determining the rate of GDP production during storage was temperature. The GDPs created by heat sterilization promoted further degradation of glucose during subsequent storage. A pH of around 3.2 protected glucose from degradation during both heat sterilization and storage. At a storage temperature of 20 degrees C and a pH of 3.2, degradation was almost negligible. Heat sterilization produced considerable amounts of GDPs absorbing at 228 nm. During initial storage, these 228 nm-absorbing GDPs almost disappeared. After reaching a nadir, absorbance at 228 nm again started to increase. Contrary to this, absorbance at 284 nm [caused mainly by 5-hydroxymethyl-2-furaldehyde (5-HMF)] increased during the whole storage period. After 2 years at 40 degrees C, the concentrations of GDPs produced during storage were of the same magnitude as those caused by heat sterilization. Inhibition of cell growth of L-929 fibroblasts correlated well with the part of the absorbance at 228 nm not caused by 5-HMF in glucose solutions that were heat sterilized under a wide range of conditions. This part of 228 nm absorbance (denoted 228corr) was caused almost entirely by 3,4 dideoxyglucosone-3-ene (3,4-DGE). CONCLUSIONS: Temperature is the single most important factor for glucose degradation during storage. The concentrations of bioincompatible GDPs produced may, under improper conditions, be as high as those produced during sterilization. High concentrations of glucose and low pH protect glucose from being degraded during both sterilization and storage. A good estimate of 3,4-DGE concentration in the fluids can be obtained correcting the UV absorbance at 228 nm for the influence from 5-HMF (and, when appropriate, for lactate). The 228corr may thus be used as a simple quality control for the fluids. PMID- 15335155 TI - PD fluids contain high concentrations of cytotoxic GDPs directly after sterilization. AB - OBJECTIVE: Glucose degradation products (GDPs) in peritoneal dialysis (PD) fluids are cytotoxic and affect the survival of the peritoneal membrane. One of the most reactive GDPs in PD fluids is 3,4-dideoxyglucosone-3-ene (3,4-DGE). 3,4-DGE has been reported as an intermediate between 3-deoxyglucosone (3-DG) and 5 hydroxymethyl furaldehyde (5-HMF) during degradation of glucose. In PD fluids, 3,4-DGE exists in a temperature-dependent equilibrium with a pool of unidentified substances. The aim of this study was to explore this equilibrium and its temperature dependence during the first months of storage after the sterilization procedure. METHODS: GDPs and inhibition of cell growth (ICG) were measured directly after sterilization of the PD fluid and during storage at different temperatures for 60 days. The following GDPs were analyzed: 3-DG, 3,4-DGE, 5-HMF, formaldehyde, acetaldehyde, glyoxal, and methylglyoxal. RESULTS: Immediately after sterilization, the concentration of 3,4-DGE was 125 micromol/L. During the first weeks of storage, it decreased by about 80%. At the same time, the 3-DG concentration increased. None of the other GDPs were significantly affected. Cytotoxicity correlated well with the concentration of 3,4-DGE. When pure 3,4-DGE was substituted for the lost amount of 3,4-DGE after 30 days of storage, the initial ICG was almost completely regained. CONCLUSIONS: Heat sterilization of PD fluids promotes the formation of large quantities of 3,4-DGE, rendering the fluid highly cytotoxic. During storage, the main part of 3,4-DGE is reversibly converted in a temperature-dependent manner to a less cytotoxic pool, consisting mainly of 3-DG. Cytotoxicity seems to be dependent exclusively on 3,4-DGE. In order to avoid higher levels of 3,4-DGE concentrations, PD fluids should not be used too soon after sterilization and should not be stored at temperatures above room temperature. PMID- 15335156 TI - Stercoral perforation of the sigmoid colon in a patient undergoing CAPD: case report. PMID- 15335157 TI - Stenotrophomonas maltophilia peritonitis in CAPD patients: susceptibility to antibiotics and treatment outcome: a report of five cases. PMID- 15335158 TI - An unusual cause of peritoneal dialysate drainage failure--inadvertent placement of Tenckhoff peritoneal dialysis catheter into an occult inguinal hernia. PMID- 15335159 TI - [The regulatory role of oxygen radicals in myocardial cells]. AB - The paper reviews participation of primary active forms of oxygen in the processes of regulation of intracellular metabolism. Action of the primary forms of oxygen upon the myocardium contractile function depends on their concentration: their moderate increase enhances the rate and strength of contractions whereas higher concentrations diminish the contraction strength by inhibiting Ca2+ inflow from without. For NO the latter effect might be compensated by absence of coronary vessels dilatation. The level of the active forms of oxygen in the cells is subsidiary to the antioxidant system, and loss of the latter's components might trigger the apoptosis process and, consequently, some diseases. PMID- 15335160 TI - [Role of mechanosensitivity of the endothelium in weakening of the vascular bed vasoconstriction]. AB - The effect of control of arterial diameter by the shear stress at the endothelium on noradrenaline-induced constriction of femoral vascular bed was investigated in anaesthetised cats. We compared noradrenaline-induced responses during the perfusion of the hindlimb at a constant blood flow and at a constant pressure as vasoconstriction is accompanied by an increase in wall shear stress only in the former case. We found that the same concentration of noradrenaline at a constant flow caused an augmentation of vascular resistance that was considerably smaller than at a constant pressure perfusion. This difference was almost eliminated after either removal of the endothelium or selective impairment of the endothelial sensitivity to the shear stress. These findings demonstrate that the control of arterial smooth muscle tone at a constant blood flow by shear stress at the endothelium does weaken noradrenaline-induced vasoconstriction. PMID- 15335161 TI - [Signaling regulatory mechanisms of the contractile activity of smooth muscle]. AB - A comparative model been designed to study a contribution of proteinkinase C (PKC)-activated intracellular signaling pathways in generation of different contractile responses of vascular (tonic) and visceral (phasic) smooth muscles. We have determined that, in tonic smooth muscle, PKC mediates activation of MAP kinases that phosphorylate key regulatory proteins of the contractile system, myosin light chain kinase and caldesmon, leading to upregulation of actomyosine motor activity. In contrast, the MAP-kinase activation is uncoupled from the contractile machinery in phasic smooth muscles, which also reveal high levels of myosin light chain kinase-related protein KRP that contributes to relaxation. Phosphorylation of KRP following activation of PKC or cyclic nucleotide-dependent protein kinases enhances the KRP activity and further contributes to relaxion in phasic smooth muscle. A possibility is discussed for exploitation of the comparative model described herein for investigation of specific role of other regulatory intracellular pathways in generation of vascular tonic contraction. PMID- 15335162 TI - [Hemodynamics mechanisms of changes in the right atrial pressure following intravenous injection of the depressor drugs]. AB - Changes of the right atrial pressure, superior and inferior vena cava flows, right ventricular myocardial contractility (first derivate of right ventricular pressure, dP/dt max) following i.v. injection of acetylcholine, histamine and isoproterenol, were studied in acute experiments on anaesthetized mongrel cats with artificial lung ventilation and opened chest. The right atrial pressure in those cases could be increased (I group of animals) or decreased (II group). In maximal shifts of right atrial pressure following acetylcholine injection, the superior vena cava flow increased but the inferior vena cava flow decreased in equal proportion. When the right ventricular myocardial contractility decreased more than the right atrial pressure was augmented, and when the cardiac negative inotropic effect was weak, the right atrial pressure was reduced. After histamine injection in both groups of animals, right ventricular myocardial contractility was increased on the same level, and changes of the inferior vena cava flow were insignificant. The right atrial pressure was elevated following greater increase of superior vena cava flow. Isoproterenol caused the positive cardiac inotropic effect and augmenting of the superior vena cava flow in both groups of animals. The right atrial pressure was elevated if the inferior vena cava flow increased and, on the other hand, when the inferior vena cava flow decreased the right atrial pressure was reduced. Thus different maximal changes of the right atrial pressure following i.v. injection of acetylcholine, histamine and isoproterenol could be explained by different hemodynamic mechanisms of the interaction between superior and inferior vena cava flow shifts and changes of the right ventricular myocardial contractility. PMID- 15335163 TI - [Participation of the cava vein blood flow in forming the total venous return under influence of different modality stimuli on the circulation system]. AB - Participation of the anterior and posterior veins cava in forming the total venous return under pressor and depressor effects, stimulation of depressing foci of the medulla's ventral part, enhancement of pulmonary ventilation, hypoxia, hypothermia, administration of acetylcholine, histamine, corinfar, was shown to depend on the blood flow shift direction in each of the veins cava, dynamics of shifts' development in time, and intensity of the stimulus. In systemic responses, the blood flow shifts in the vena cava anterior much contribute to the total venous return at the maximum of the systemic arterial pressure rise (r = 0.87) whereas contribution of the vena cava posterior is the greatest during a later occurring increase in the venous return (r = 0.84). Along with increase in the stimulus intensity the vena cava anterior's part in forming the venous return becomes more limited whereas that of the vena cava posterior is enhanced. PMID- 15335164 TI - [The dynamics of leukocyte adhesion to the endothelium pial venules under massive blood loss]. AB - The dynamics of leukocytes adhesion to brain microvenules endothelium of Wistar rats were studied under acute massive blood loss. Using the method of vital videomicroscope study of the influence of the influence of the blood volume reduction on the leukocyte-endothelium interaction is investigated. We studied vessels of the diameter under 30 mcm. It is shown that under progressing reduction of the system arterial pressure under 60 mm Hg, the number of leukocytes adhesion to venules' wall reached 20 +/- 3 on 100 mcm, but in terminal stage such acts are the greatest. It is supposed that as a result massive blood loss leukocytes turn out to be the direct reason of the deep pathological disturbances of microcirculation in the brain. PMID- 15335165 TI - [Circulation changes depending on manifestations in soft tissue tensioning in the leg lengthening process]. AB - Surgical intervention was found to intensify circulation in the limb and redistribute the blood flow. Leg lengthening led to arterial inflow limitation due to the magisterial artery strain. The changes were accompanied both by increase of functioning capillaries in number and increase of venous outflow dependence on blood inflow. The decreased after surgery oxygen tension in m. gastrocnemius did not change throughout distraction. The increase of functioning capillaries in number in resting contributed to maintenance of tissue oxygenation in the initial period of distraction, and in case of maximal tissue tensioning hydrostatic pressure increased in the capillaries due to arterial pressure rise. Restoration of the circulation parameters in the fixation period started with an increase of circulation volumetric rate in vessels with preservation of the rest mechanisms of the tissue oxygenation maintenance. PMID- 15335166 TI - [Features of the spontaneous unit activity in the human thalamic parafascicular (CM-Pf) complex and its modification in functional cerebral changes]. AB - New data on neural organization of the human thalamic parafascicular (CM-Pf) complex were revealed with microelectrode technique during 11 stereotaxic operations in alert diskinetic patients suffering from the tonic forms of spasmodic torticollis. The data were obtained as follows: the functional heterogeneity in cellular organization of the human CM-Pf thalamic nuclei and the existence of three (A, B and C) different types of neurons in these thalamic nuclei: with the irregular discharges (A-type, 18%); with short (10-20 ms) bursts characterised by unstable rhythmic 2-5 Hz pattern and by the low threshold Ca2+ dependent K+ conductance (B-type, 77%); with long-lasting (0.1-2.0 s) bursts of high-frequency trains and constant interburst intervals (C-type, 5%). The functional cerebral changes after motor test performances were shown to be accompanied by appearance of transient modifications of the background unit activity pattern and by tendency towards an increase of neural activity local synchronisation with some stabilising of oscillatory rhythm of discharging B-type neurons. For the first time, a direct relationship between functional characteristics of the human thalamic CM-Pf units and the motor deviations was found in spasmodic torticollis patients. PMID- 15335167 TI - [Psychosomatic factors of resistance to hypoxia]. AB - During formation of human long-term adaptation to cold-hypoxic influence, the some dynamics of psychological mechanisms contribution was revealed. On the first step of adaptation process, emotional-volitional personal peculiarities are significantly contributing; on further steps, communicability and general learning abilities are gaining importance. Anxiety, tenseness, diffidence and too strong self-control limit the hypoxic resistance far before restriction of physiological markers of the organism's functional abilities. Adaptation to cold hypoxic effect contributing to organism stress-limiting system is believed to help experiencing extreme somatic and psychic loads by the people from psychological risk group. PMID- 15335168 TI - [The role of glucocorticoid hormones in interleukin-1 signal transduction via the sphingomyelin pathway]. AB - Interleukin-1 and glucocorticoid hormones are the key transmitters of interaction between the neuroendocrine and immune systems. To study the molecular mechanisms of immunomodulatory effects of Interleukin-1 and glucocorticoid hormones, a search for changes in activity of neutral sphingomyelinase: the main marker of initiation of Interleukin-1beta signal transduction via the sphingomyelin pathway in target cells, was accomplished. The Interleukin-1beta was found to activate neutral sphingomyelinase both in P2 fraction of murine brain cortex and membranes of immune-competent cells. Experimental modifications of endogenous glucocorticoid level in the mouse blood were for the first time shown to induce changes in neutral sphingomyelinase activity in membranes of the cells of the immune and nervous systems. It appears that the sphingomyelinase pathway of Interleukin-1beta signaling might be a possible target for glucocorticoid hormones' immune-modulating effects. PMID- 15335169 TI - [Heterogeneity of molecular mechanisms the olfactor reception]. AB - In experiments on the frog isolated olfactory epithelium by using vital fluorescent microscope, odorants with fruit, rank, flower and camphor smell were shown to involve intracellular signaling systems in olfactory transduction. The odorants with different qualitative smells have different messenger and activity mechanisms. Intracellular messengers do not participate in reception of odorants with piquant and rotten smells. Thus the perception of different odour substances is maintained by physical and chemical processes. Hence, not only taste, carotid, medullar, but olfactory reception as well are characterised by heterogeneity of biophysical mechanisms. PMID- 15335170 TI - [The modulating action of amino acids upon development in an organotypic culture]. AB - The effect of L-amino acids in concentration 0.05 ng/ml was investigated in organotypic tissue culture of spleen, liver and brain cortex at the age of 1 and 21 days. The high molecular and low hydrophobic amino acids: asparagin, lysine, arginine, and glutamine, are active inducing both a less intensive growth zone in explants of all three tissue types in 1-day old rats and more intensive growth zone in spleen, liver explants in 21-day rats as compared to control. Another group of low molecular and high hydrophobic amino acids is active inducing a more intensive growth zone in brain cortex explant in 21-day rats, as compared to control. The data obtained suggest a modulating role of amino acids in the tissues at different stages of maturation. PMID- 15335171 TI - [The effect of chronic experience of victories or defeats in social conflicts upon sexual motivation in male mice]. AB - The relevance of using the social contact test "Peregorodka" for investigation into sexual motivation in male mice, was checked and confirmed. A positive correlation was found between behavioural patterns manifested by the males near a transparent septum with holes separating them from receptive females and the males' subsequent sexual behaviour. Experience of victories as well as defeats in social conflicts was found not to interfere with occurrence of sexual motivation of the same intensity as in control animals. In the latter, however, sexual motivation was stable in time whereas in the model males its gradual decrease occurred within the 30-min test period. The findings suggest fast exhaustion of the sexual motivation in mice under conditions of prolonged social conflicts irrespective of either victories or defeats. PMID- 15335172 TI - Obstructive sleep apnea-hypopnea syndrome and cardiovascular diseases. PMID- 15335173 TI - Under-use of warfarin for patients with non-valvular atrial fibrillation in Japan. PMID- 15335174 TI - Double therapy-related leukemia: coincidental occurrence or susceptibility for leukemia? PMID- 15335175 TI - Photosensitive drug eruption induced by efavirenz in a patient with HIV infection. PMID- 15335176 TI - The role of bacterial infection in the pathogenesis of inflammatory bowel disease. AB - In the last decade, the dogma that no bacteria could grow in the acid milieu of the stomach has been destroyed by evidence that the infective agent, H. pylori, is responsible for gastric and duodenal disease. Studies on H. pylori infection suggest that some strains of intestinal bacteria may be responsible for intestinal ulceration and inflammation concomitant with inflammatory bowel diseases (IBD), i.e., ulcerative colitis and Crohn's disease. Evidence for pathophysiological roles for certain strains of luminal bacteria result from a number of IBD animal models. Recent studies on innate immunity, including toll like receptors and NOD isoforms, suggest that bacterial infections may contribute to intestinal inflammation in genetically susceptible hosts. This brief review focuses on the bacterial pathogenesis and the role of innate immunity in the etiology of IBD's. PMID- 15335177 TI - Myelofibrosis with myeloid metaplasia: new developments in pathogenesis and treatment. AB - Myeloid metaplasia with myelofibrosis (MMM) is a chronic myeloproliferative disorder (CMPD) characterized by progressive anemia, massive splenomegaly, both hepatosplenic and non-hepatosplenic extramedullary hematopoiesis (EMH), a leukoerythroblastic blood smear, circulating progenitor cells, and marked bone marrow stromal reaction including collagen fibrosis, osteosclerosis and angiogenesis. The overall median survival is 5 years although it might range from 2 to 15 years depending on the presence or absence of clinically defined prognostic factors. Death is often due to leukemic transformation, portal hypertension or infection. In addition to shortened survival, quality of life is often affected by frequent red blood cell transfusions, profound constitutional symptoms, and cachexia. Drug therapy and autologous hematopoietic stem cell transplantation (HSCT) are of only palliative value and have not been shown to improve survival. The role of allogeneic HSCT, both myeloablative and non myeloablative, is actively being investigated. Both splenectomy and radiation therapy have defined therapeutic roles to control EMH-associated symptoms. Analysis of the molecular biology of the disease is underway with the aid of animal models leading to the identification of novel therapeutic targets. Among the novel agents tested, thalidomide seems the most promising although newer agents are on the horizon. PMID- 15335178 TI - Comparison of serum IgA and IgG antibodies for detecting Helicobacter pylori infection. AB - OBJECTIVE: Although the diagnostic utility of serum IgG antibodies to Helicobacter pylori (H. pylori) is well established, the usefulness of IgA-based tests is less well documented. The aim of this study was to evaluate two commercially available ELISAs, both for IgG and IgA. PATIENTS AND METHODS: Rapid urease test and histology analysis were performed in 183 patients. A patient was considered to be H. pylori-positive when either biopsy test was positive, and considered to be noninfected when both tests were negative. Intestinal metaplasia was determined by dye endoscopy with methylene blue. ELISA testing was performed using the EPI HM-CAP IgG and PP-CAP IgA assays and EIAgen IgG and IgA assays. RESULTS: Sensitivity was 94.7, 93.9, 94.8, and 97.0% for HM-CAP IgG, PP-CAP IgA, EIAgen IgG, and EIAgen IgA, respectively. Although sensitivity was excellent for both IgG and IgA antibodies, specificity of both IgA EIAs was low (PP-CAP 72.6%, EIAgen H. pylori IgA 59.2%). Three of 101 H. pylori-infected patients were PP-CAP positive and HM-CAP negative and four were EIAgen H. pylori IgA positive and EIAgen IgG negative. Of eight noninfected patients in whom intestinal metaplasia was found, PP-CAP IgA results were positive in three of five patients with a HM CAP IgG negative result and EIAgen IgA was detected in one of four patients with an EIAgen IgG negative result. CONCLUSIONS: Since some patients have IgA positive but IgG negative results, great care should be taken not to underestimate the prevalence of H. pylori infection from the results of IgG serology. PMID- 15335180 TI - Effects of the long-acting calcium channel blockers, amlodipine, manidipine and cilnidipine on steroid hormones and insulin resistance in hypertensive obese patients. AB - OBJECTIVE: To demonstrate that calcium channel blockers can improve insulin resistance clinically, we investigated the effects of the calcium channel blockers, amlodipine, manidipine and cilnidipine on serum levels of steroid hormones and insulin. SUBJECTS AND METHODS: Thirty hypertensive obese patients [15 men and 15 women; mean age 55.9 years, mean body mass index (BMI) 27.6] were divided into three groups and treated with either 5 mg of amlodipine, 20 mg of manidipine or 10 mg of cilnidipine. Blood pressure (BP), fasting plasma glucose (FPG), HbA1c, fasting serum immunoreactive insulin (F-IRI), insulin resistance index [as assessed by the homeostasis model assessment (HOMA-R)], serum DHEA, serum DHEA-S, plasma ACTH, serum cortisol, plasma renin activity (PRA), and serum aldosterone, were measured before and after 1, 2, 3 and 6 months of treatment. RESULTS: In all three groups, BP decreased significantly after 1 month and F-IRI and HOMA-R decreased significantly after 2-3 months. A concurrent rise in serum DHEA and DHEA-S levels was also observed, however, the differences were not significant. No changes in FPG, HbA1c, ACTH, cortisol, PRA or aldosterone levels were observed during treatment. CONCLUSIONS: We conclude that amlodipine, manidipine and cilnidipine all improve insulin resistance and consequently increase serum levels of DHEA and DHEA-S. PMID- 15335179 TI - Physicians' attitudes toward anticoagulant therapy in patients with chronic atrial fibrillation. AB - INTRODUCTION: Although many clinical trials have demonstrated that anticoagulant therapy substantially reduces the risk of ischemic stroke in patients with atrial fibrillation (AF), some physicians are reluctant to use anticoagulants. We investigated attitudes of physicians in Japan toward anticoagulant therapy in chronic AF patients. METHODS: We conducted a survey at the annual meeting of the Japanese Society of General Medicine. We presented subject physicians with 8 vignettes of chronic AF patients and requested that they indicate their most favored choice of therapy from among 6 strategies including warfarin and aspirin. RESULTS: We distributed 209 questionnaires and received 139 replies (67% response rate). For all 8 vignettes presented, only 26% of the respondents preferred to use anticoagulant therapy in AF patients. Longer clinical experiences and responsibility at a teaching hospital were associated with negative attitude toward anticoagulant therapy, while experience of preventive therapy in patients with thromboembolism due to AF and strong influence of clinical trials of anticoagulant prophylaxis on their practice were associated with positive attitude toward the therapy. Among patient characteristics in the vignettes, a risk of thromboembolism was positively associated with preference for anticoagulant therapy, but an advanced age and a risk of bleeding complications were negatively associated with the preference for the therapy. CONCLUSIONS: The physicians in Japan in this survey, especially those with longer clinical experiences or responsibility at a teaching hospital, have a negative attitude toward anticoagulant therapy in chronic AF patients. An advanced age and a risk of bleeding complications of patients are deterrent factors to the use of anticoagulant therapy. PMID- 15335181 TI - Whipple's disease: the first Japanese case diagnosed by electron microscopy and polymerase chain reaction. AB - A 52-year-old man presented with diarrhea and 20 kg weight loss in one year. Enteroscopy showed diffuse yellow-white shaggy mucosa in the duodenum and jejunum. Biopsies of the duodenal mucosa revealed massive infiltration within the lamina propria by foamy macrophages strongly positive for periodic acid-Schiff stain. Electron microscopy demonstrated numerous bacilli within macrophages of the lamina propria. Tropheryma whipplei DNA was detected by polymerase chain reaction. The definitive diagnosis of Whipple's disease was made. Antibiotic therapy dramatically improved his clinical picture. This is the first Japanese case with Whipple's disease diagnosed by electron microscopy and polymerase chain reaction. PMID- 15335182 TI - Adult-onset idiopathic hypogonadotropic hypogonadism due to isolated pituitary gonadotropin deficiency. AB - A 25-year-old Japanese man with adult-onset idiopathic hypogonadotropic hypogonadism is reported. He had been delivered normally, had normal puberty, and experienced erectile dysfunction at age 24 years. Brain MRI revealed no abnormal findings and endocrinological data supported the diagnosis of isolated gonadotropin deficiency. Although most patients with idiopathic hypogonadotropic hypogonadism have a hypothalamic dysfunction, the lesion in this case may be considered to be in the pituitary since repetitive GnRH loading failed to increase serum LH and FSH. PMID- 15335183 TI - Transient appearance of lactate dehydrogenase (LDH)-linked immunoglobulin and thyroid dysfunction at the postpartum period. AB - Here, we report a 28-year-old woman who transiently showed lactate dehydrogenase (LDH)-linked immunoglobulin during postpartum thyroiditis. She demonstrated high levels of serum LDH (794 IU/l) and thyroid hormones 7 months after delivery. Electrophoretic isoenzyme analysis of LDH showed an abnormal broadband caused by LDH-linked immunoglobulin (IgG-kappa). Transient thyrotoxicosis due to postpartum thyroiditis improved without any specific treatment, and elevated serum concentration of LDH decreased to the normal level (395 IU/l) with disappearance of LDH-linked IgG. LDH-linked immunoglobulin may also appear at the postpartum period. PMID- 15335184 TI - Peripheral primitive neuroectodermal tumor of the chest wall of a 69-year-old man. AB - We report a case of peripheral primitive neuroectodermal tumor (pPNET), which belongs to the pPNET/Ewing's sarcoma family, arising in the chest wall of a 69 year-old man. He had high levels of serum neuron-specific enolase and pro-gastrin releasing peptide, which are believed to be useful diagnostic blood markers for small cell lung carcinoma (SCLC). Microscopically, the tumor was composed of solid nests and sheets of monotous, primitive, small round cells with a few rosettes, making it difficult to distinguish from SCLC. Immunohistochemically, the tumor cells showed intense cell membranous immunoreactivity for MIC2 protein (CD99). EWS/FLI-1 chimeric mRNA that originated from the characteristic t(11;22)(q24;q12) chromosomal translocation was detected by RT-PCR and nucleotide sequence analysis. These results confirmed the diagnostic validity of the present tumor being a pPNET, thus raising the possibility that in the past, pPNETs which have arisen in the chest have been mistakenly diagnosed as SCLC. PMID- 15335185 TI - Therapy-related myelodysplastic syndrome with trisomy 1q due to der(1;7) and megakaryoblastic proliferation developing during complete remission of therapy related acute myeloid leukemia with t(8;21). AB - Therapy-related acute myeloid leukemia (t-AML) with t(8;21) and therapy-related myelodysplastic syndrome (t-MDS) with trisomy 1q due to der(1;7) developed in the same patient with T-cell lymphoma at intervals of six years. After the development of t-MDS with trisomy 1q, during complete remission of t-AML, the number of megakaryoblasts increased to maximally 74% of leukocytes in the blood. This is a very rare case of two separate therapy-related myeloid malignancies (early t-AML and late t-MDS) and is also a notable case of t-MDS with trisomy 1q due to der(1;7) accompanied by megakaryoblastic proliferation. PMID- 15335186 TI - Heparin-induced thrombocytopenia and thrombosis in a patient with polycythemia vera. AB - A 75-year-old Japanese woman with polycythemia vera was admitted to our hospital in January 2003 with suspected pulmonary thromboembolism. After administration of heparin, platelet count decreased from 1,694 x 10(9)/l on admission to 60 x 10(9)/l on hospital day 14. The patient developed acute limb embolism and transient cerebral ischemic attack on days 17 and 25, respectively. Signs and symptoms mimicked those of disseminated intravascular coagulation. Antibodies against heparin and platelet factor 4 complexes were detected in serum, and a diagnosis of heparin-induced thrombocytopenia and thrombosis was made. Argatroban treatment improved thrombocytopenia and hypercoagulable state. PMID- 15335187 TI - Non-secretory multiple myeloma with azurophilic granules and vacuoles: an immunological and ultrastructural study. AB - Cytoplasmic vacuolation was seen in patients with a variety of plasma cell dyscrasia. We report here a case of leukemic non-secretory multiple myeloma with many azurophilic granules. By electron microscopy, the myeloma cells were found to have a well-developed rough endoplasmic reticulum and a clear Golgi apparatus, and azurophilic granules were identified as phagocytic vacuoles. In addition to myeloma markers, the cells were positive for B cell-associated, myeloid and stem cell markers. The diagnosis is difficult because of its misleading morphology and unusual surface markers. We consider that electron microscopy is useful for the identification of cell lineage in this disorder. PMID- 15335188 TI - Multiple myeloma complicated by autoimmune hemolytic anemia. AB - A 57-year-old man was admitted with severe anemia and hypergamma globulinemia. After a diagnosis of multiple myeloma and autoimmune hemolytic anemia was made, chemotherapy rapidly decreased the M-protein level and improved his anemia with normalization of the direct Coombs test. The immunoglobulin binding to the patient's red cells was immunoglobulin G kappa chain like the myeloma M-protein. However, monoclonal immunoglobulin G derived from short-term culture of the patient's bone marrow mononuclear cells did not bind to a panel of red cells. Therefore, the relationship between the M protein produced by his myeloma cells and hemolysis remained unclear. PMID- 15335189 TI - Focal myositis in monozygotic twins. AB - Focal myositis is a rare disease with unknown etiology and a broad spectrum. Here, we present two cases in monozygotic twins who complained of recurrent pain of their calves and showed histological signs of inflammation and MRI image compatible with the diagnosis of focal myositis. The occurrence of twin cases not living in the same household suggests a genetic susceptibility to the disease. PMID- 15335190 TI - Paraneoplastic cerebellar degeneration (PCD) associated with squamous cell carcinoma of the lung. AB - We report a case of a 71-year-old man who presented with cerebellar dysfunction. He was diagnosed as having squamous cell carcinoma of the lung (T2N3M0, Stage IIIB). No anti-onconeural antibodies were found in his serum. Cerebral spinal fluid (CSF) examination showed mild mononuclear pleocytosis alone. Magnetic resonance imaging (MRI) of the brain and spinal cord revealed no abnormalities. At autopsy, there was complete disappearance of Purkinje cells with reactive astrocytosis. These findings are compatible with paraneoplastic cerebellar degeneration (PCD). To our knowledge, no case of PCD has been reported previously in patients with squamous cell carcinoma of the lung. PMID- 15335191 TI - Deterioration of vascular dementia caused by recurrent multiple small emboli from thoracic aortic atheroma. AB - We report a case of a 77-year-old man with deteriorating dementia caused by repeated multiple small cerebral embolisms from a thoracic aortic atheroma. Multiple small embolisms were confirmed by diffusion-weighted magnetic resonance imaging (DWI). The patient ultimately died due to aortic dissection. Pathological examinations revealed that no causative embolic source for multiple embolisms could be detected other than severe atheromatous ulcer in thoracic aorta. This case demonstrates that severe aortic atheroma has the potential to precipitate deterioration of vascular dementia. PMID- 15335192 TI - The efficacy of interferon-alpha in a patient with resistant familial Mediterranean fever complicated by polyarteritis nodosa. AB - Familial Mediterranean fever (FMF) is a recurrent self-limiting polyserositis. Polyarteritis nodosa (PAN) complicating FMF is very rare. Here, we present a 17 year-old male patient with FMF who subsequently developed PAN 2 weeks after hepatitis A infection. This case was also complicated with perirenal haematoma, and right nephrectomy was performed. The clinical condition of the patient was improved after therapy with intravenous and oral corticosteroid and intravenous cyclophosphamide. However, the FMF attacks and vasculitic skin lesions again occurred while he was using colchicine plus immunosuppressive agents a few months later. Interferon-alpha therapy was administered and the attacks were resolved within 3 months. He has not experienced any other symptom during the follow-up period of 28 months. PMID- 15335193 TI - Pyothorax caused by Nocardia otitidiscaviarum in a patient with rheumatoid vasculitis. AB - We report a case of pyothorax caused by Nocardia (N.) otitidiscaviarum infection in a 69-year-old man with rheumatoid vasculitis, who was regularly treated with prednisolone in our hospital. Initially, the patient responded poorly to intravenous imipenem/cilastatin (IPM/CS), minocyclin (MINO), and oral trimethoprim-sulfamethoxazole (TMP-SMX), but later improved after treatment with levofloxacin (LVFX) and gentamicin sulfate (GM) according to in vitro susceptibility tests. To our knowledge, this is the first description of pyothorax caused by N. otitidiscaviarum infection. It is a rare disease, but recognition of the disease in immunocompromised patients and the prompt initiation of appropriate treatments based on isolation of the pathogen and susceptibility testing can lead to a successful outcome. PMID- 15335194 TI - Hemolytic uremic syndrome associated with Shiga toxin producing Escherichia coli infection in a healthy adult woman. AB - A 49-year-old healthy Japanese woman presented with hemorrhagic diarrhea because of Shiga toxin producing Escherichia coli infection, and then hemolytic uremic syndrome (HUS) developed in the patient. She was successfully treated with continuous hemodiafiltration, plasma exchange, and endotoxin adsorption therapy. An analysis of previous case reports suggests that females aged between 16 and 65 years are at an increased risk of HUS resulting from hemorrhagic colitis. We propose that adult female patients with hemorrhagic colitis should be carefully monitored regardless of their medical history, physical presentation, or laboratory data. PMID- 15335195 TI - Quinine-resistant severe falciparum malaria effectively treated with atovaquone and proguanil hydrochloride combination therapy. AB - A 22-year-old Japanese man noticed pyrexia and diarrhea after travel to Guinea. Notable physical findings included hepatosplenomegaly. Treatment with oral quinine and minocycline was started after definitive diagnosis of falciparum malaria by blood smear. Initially, parasitemia and body temperature decreased but by the third night of therapy his temperature increased to 40 degrees C with a slight increase of parasite count. When quinine treatment was changed to atovaquone/proguanil, his temperature dropped immediately and complete plasmodial elimination was confirmed on microscopic examination. Subsequent recrudescence of the disease was not observed. It was concluded that the antimalarial treatment with atovaquone/proguanil might become invaluable in Japan. PMID- 15335196 TI - Plasma adrenomedullin concentration patients with Graves' disease. PMID- 15335197 TI - The first case of efavirenz-induced photosensitivity in a Japanese patient with HIV infection. PMID- 15335198 TI - Fitz-Hugh-Curtis syndrome: hepatic capsular enhancement and diffuse gallbladder wall thickening on contrast-enhanced CT. PMID- 15335199 TI - Multidetector-row CT imaging of aneurysm rupture of the aortic arch. PMID- 15335200 TI - Ectopic calcinosis of the mitral valve in a patient undergoing long-term hemodialysis. PMID- 15335201 TI - Flaming cells in IgG myeloma. PMID- 15335202 TI - Diffusion MRI in acute carbon monoxide poisoning. PMID- 15335203 TI - Acute gout attack in the wrist joint. PMID- 15335204 TI - Using ANOVA to analyze microarray data. AB - ANOVA provides a general approach to the analysis of single and multiple factor experiments on both one- and two-color microarray platforms. Mixed model ANOVA is important because in many microarray experiments there are multiple sources of variation that must be taken into consideration when constructing tests for differential expression of a gene. The genome is large, and the signals of expression change can be small, so we must rely on rigorous statistical methods to distinguish signal from noise. We apply statistical tests to ensure that we are not just making up stories based on seeing patterns where there may be none. PMID- 15335205 TI - AFLP analysis using GeneMapper software and an Excel macro that aligns and converts output to binary. PMID- 15335206 TI - Molecular cytogenetic characterization by combined COBRA-FISH and fiber-FISH: application to PER.C6 cells. PMID- 15335207 TI - Identifying adaptor contamination when mining DNA sequence data. PMID- 15335208 TI - Liquid gel amplification of complex plasmid libraries. PMID- 15335209 TI - Coupling homologous recombination with growth selection in yeast: a tool for construction of random DNA sequence libraries. PMID- 15335210 TI - Bioluminescence-based ATP assays using a charge-coupled device imaging system. PMID- 15335211 TI - Infrared video thermography: a technique for assessing cold adaptation in insects. AB - Insects can survive subzero temperatures by two main strategies: freeze tolerance and freeze avoidance. An array of techniques have been used to investigate the physiological limits of insects to low temperatures, such as differential scanning calorimetry, temperature-controlled cooling apparatus, thermocouples, and computer-controlled chart recording equipment. However, these techniques require animals to be stationary, precluding behavioral data. We used infrared video thermography to investigate cold adaptation in an alpine insect, expanding such investigations to include behavioral response as an indicator of physiological stress. This technique is noninvasive and provides a large amount of physiological information, such as supercooling points, lower lethal temperatures, and hemolymph melting points. Insect supercooling points in response to a constant cooling rate were variable; however, temperatures at the initiation of behavioral stress response were less variable. Assessments of supercooling points and lower lethal temperatures obtained in this way are more biologically meaningful because allowing unhindered movement of insects more closely resembles natural environments. PMID- 15335212 TI - PathoGene: a pathogen coding sequence discovery and analysis resource. AB - PathoGene is a web-based resource that streamlines the process of predicting genes in microorganisms and designs PCR primers for amplification to facilitate sequence analysis and experimentation. PathoGene currently supports primer design for every complete microbial, viral, and fungal genome as cataloged in GenBank by the National Center for Biotechnology Information (NCBI; http://www.ncbi.nlm.nih.gov/). The resulting primers can then be subjected to a stand-alone Basic Local Alignment Search Tool (BLAST) system called PathoBLAST in which the predicted PCR product and/or primers can be compared against the genome of interest or a similar genome to find related genes or estimate primer quality. PMID- 15335213 TI - Data processing and exchange tools to facilitate chemical genetic screening processes. AB - Due to the development of chemical genomics, the screening of chemical libraries is used more and more by research laboratories to identify small molecule inhibitors or activators of cell functions. To facilitate the treatment and archiving of screening data, we developed a multiuser web application called Elisa Data Exchanger (EDE). The program is able to automatically identify which chemical compounds were tested. Several data exchange formats can be generated for visualization, printing, charting, or exporting to chemical analysis software. These data exchange functions allow for a comparison of results obtained from screening several targets in order to select the most specific compounds. EDE is freely available online at https://ibph.pharma.univ montpl.fr/ede/ (login: evalede, password: loginede). PMID- 15335214 TI - AutoDimer: a screening tool for primer-dimer and hairpin structures. AB - The ability to select short DNA oligonucleotide sequences capable of binding solely to their intended target is of great importance in developing nucleic acid based detection technologies. Applications such as multiplex PCR rely on primers binding to unique regions in a genome. Competing side reactions with other primer pairs or template DNA decrease PCR efficiency: Freely available primer design software such as Primer3 screens for potential hairpin and primer-dimer interactions while selecting a single primer pair. The development of multiplex PCR assays (in the range of 5 to 20 loci) requires the screening of all primer pairs for potential cross-reactivity. However, a logistical problem results due to the number of total number of comparisons required. Comparing the primer set for a 10-plex assay (20 total primer sequences) results in 210 primer-primer combinations that must be screened. The ability to screen sets of candidate oligomers rapidly for potential cross-reactivity reduces overall assay devlelopment time. Here we report the application of a familiar sliding algorithm for comparing two strands of DNA in an overlapping fashion. The algorithm has been employed in a software package wherein the user can compare multiple sequences in a single computational run. After the screening is completed, a score is assigned to potential duplex interactions exceeding a user-defined threshold. Additional criteria of predicted melting temperature (Tm) and free energy of melting (deltaG) are included for further ranking. Sodium counterion and total stand concentrations can be adjusted for the Tm and deltaG calculations. The predicted interactions are saved in a text file for further evaluation. PMID- 15335215 TI - Parameters affecting substance P measurement in heart, lung, and skin. AB - Substance P (SP), a neuropeptide that is widely distributed both peripherally and centrally, mediates several pathophysiological processes. Among current assays for SP, enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) have been most widely used. Several previous studies, mostly performed with nerve extracts or organ perfusates, determined that acidity of the extraction buffer as well as the number extractions performed constitute factors influencing accurate measurements. We used an ELISA protocol in this study to analyze methodological aspects of SP measurement in extracts from heart, skin, and lung. The extraction procedure had two steps, an acid extraction followed by a column extraction. We could effectively measure SP with extract from as little as 10 mg of tissue. For each tissue examined, different variables influenced the SP measured. For all tissues, the weight of tissue extracted was critical; the more tissue extracted, the lower the sensitivity of the assay. This problem could be overcome in skin by omitting the column extraction. When mechanical loses were considered (e.g., loss during extraction and SP retained by the column after elution), column extraction improved SP measurements only with lung tissue. The amount of SP remaining in the sample after the first extraction also varied among tissues. The first acid extraction effectively isolated 80% of total SP from skin. In contrast, the first extraction with lung tissue recovered only 58%. Because both acid and heat effectively release SP from nerve endings, this could reflect the presence of non neuronal SP, especially in lung. High-dose capsaicin treatment, which depletes SP in nerve endings, caused 42% loss of SP in skin independent of amount of tissue extracted Our results suggest that a second acid extraction of tissue should be performed and that column extraction is clearly detrimental with skin samples. PMID- 15335216 TI - Potential influence of the first PCR cycles in real-time comparative gene quantifications. AB - There is an underlying assumption in real-time PCR that the amplification efficiency is equal from the first cycles until a signal can be detected. In this study, we evaluated this assumption by analyzing genes with known gene copy number using real-time PCR comparative gene quantifications. Listeria monocytogenes has six 23S rRNA gene copies and one copy of the hlyA gene. We determined 23S rRNA gene copy numbers between 0.9 and 1.6 relative to hlyA when applying the comparative gene quantification approach. This paper focuses on the first cycles of PCR to explain the difference between known and determined gene copy numbers. Both theoretical and experimental evaluations were done. There are three different products (types 1-3) dominating in the first cycles. Type 1 is the original target, type 2 are undefined long products, while type 3 are products that accumulate during PCR. We evaluated the effects of type 1 and 2 products during the first cycles by cutting the target DNA with a restriction enzyme that cuts outside the boundaries of the PCR products. The digestion resulted in a presumed increased amplification efficiency for type 1 and 2 products. Differences in the amplification efficiencies between type 1, 2, and 3 products may explain part of the error in the gene copy number determinations using real-time PCR comparative gene quantifications. Future applications of real time PCR quantifications should account for the effect of the first few PCR cycles on the conclusions drawn. PMID- 15335217 TI - Phage matrix for isolation of glioma cell membrane proteins. AB - Cell-binding ligands for RG2 rat glioma were identified in our recent study from a library of peptides that are displayed as fusion molecules on phage particles. Here, one of the phage clones was used to affinity purify those cell membrane components to which the displayed peptides bind. This phage clone, displaying the ELRGDSLP peptide, was shown to recognize glioma cells specifically in comparison to control phage-expressing peptides of either similar or irrelevant sequences. Blocking experiments with synthetic RGDS peptide demonstrated that the phage glioma cell recognition occurs via the RGD motif known to be present in many integrin-binding proteins. To form an affinity matrix that would bind to glioma cell membrane molecules, ELRGDSLP phage particles were cross-linked using dextran polymer. Whole cell lysate from RG2 rat glioma cells was passed through the matrix, resulting in the isolation of cell membrane components having strong affinity to the peptides on phage and molecules associated with those components. One of the isolated proteins was found to be CD44s, a cell surface adhesion molecule involved in glioma cell invasion and migration, which likely formed a complex with an RGD-binding integrin. Cell membrane proteins isolated with this innovative approach could be used for the design of cell-specific anticancer treatments. PMID- 15335218 TI - Fresh and cultured buccal cells as a source of mRNA and protein for molecular analysis. AB - We developed a method for obtaining viable buccal cells from mouthwash samples for use as a source of mRNA and protein. Immunofluorescent analysis showed that most cells were derived from nonkeratinized parabasal epithelia, with a minor proportion of proliferative cells. Gene expression was detected in buccal cells using reverse transcription PCR, Western blot analysis, and immunofluorescence. Using a keratinocyte-specific medium, buccal cells could be cultured on Matrigel coated permeable filters for up to 2 weeks while maintaining the expression of some epithelial-specific markers, including cytokeratin 13, cytokeratin 10, transferrin receptor, and beta-integrin. The basal marker cytokeratin 14 and Ki67, an indicator of cellular proliferation, were detected in a few cells. We show that buccal cells can be obtained from a noninvasive procedure for use as a source of material for biochemical analyses. A population of the buccal cells can be maintained in culture for up to 2 weeks using keratinocyte-specific medium in combination with extracellular matrix. PMID- 15335219 TI - Transduction of multiple cell types using improved conditions for gene delivery and expression of SV40 pseudovirions packaged in vitro. AB - This comprehensive study demonstrates highly efficient transduction of a wide variety of human, murine, and monkey cell lines, using a procedure for in vitro packaging of plasmid DNA in recombinant simian virus 40 (SV40) capsid proteins to form pseudovirions. The pseudovirions are encapsidated by the VP1 major capsid protein, with no SV40 sequence requirement, and are able to carry up to 17.7 kb of supercoiled plasmid DNA. We developed a procedure to scale-up production of SV40 pseudovirions, as well as an efficient protocol to concentrate the virions with no loss of activity. We also developed a method that allows transduction of 10 times more cells than the original protocol. This protocol was tested using supercoiled in vitro-packaged plasmid carrying the human multidrug-resistance gene (MDR1 encoding P-glycoprotein; P-gp), or the enhanced green fluorescent protein reporter gene (EGFP) in .45 human lymphoblastoid cells and in K562 human erythroleukemia cells. Multiple transductions at 24-h intervals were shown to increase expression using the EGFP reporter gene. The protocols developed in this study establish in vitro-packaged SV40 pseudovirions as one of the most efficient gene delivery systems. PMID- 15335220 TI - Production of stably transfected cell lines using immunoporation. AB - Previous work from this laboratory has shown that immunoporation has the potential for the selective transfection of a range of different animal cells based on their immunological identity. The unique ability of immunoporation to target cells for transfection combined with the high efficiency of transfection and the high viability of cells make this method extremely promising for scientific and medical research. The experiments reported here show that not only can immunoporation produce transient transfection but also stably transfected cells are produced and such types of cells will be essential for the use of this method for gene therapy. PMID- 15335221 TI - Creation of a minimal tiling path of genomic clones for Drosophila: provision of a common resource. AB - On the basis of shotgun subclone libraries used in the sequencing of the Drosophila melanogaster genome, a minimal tiling path of subclones across much of the genome was determined. About 320,000 shotgun clones for chromosomes X(12-20), 2R, 2L, 3R, and 4 were available from the Berkeley Drosophila Genome Project. The clone inserts have an average length of 3.4 kb and are amenable to standard PCR amplification. The resulting tiling path covers 86.2% of chromosome X(12-20), 86.2% of chromosomal arm 2R, 79.0% of 2L, 89.6% of 3R, and 80.5% of chromosome 4. In total, the 25,135 clones represent 76.7 Mb--equivalent to about 67% of the genome--and would be suitable for producing a microarray on a single slide. PMID- 15335222 TI - Characterization of an inexpensive, nontoxic, and highly sensitive microarray substrate. AB - An agarose film has been proposed as an efficient substrate for producing microarrays. The original film preparation procedure was simplified significantly by grafting the agarose layer directly onto unmodified microscope glass slides instead of aminated glass slides, and the blocking procedure was replaced with a wash in 0.1x standard saline citrate (SSC) and 0.5% sodium dodecyl sulfate (SDS) without decreasing the performance of the produced microarrays. Characterization of the grafted agarose film using atomic force microscopy (AFM) and scanning electron microscopy (SEM) showed that the agarose film had a 10-fold increase in surface roughness compared to glass and that the interior of the agarose film was porous, with pore sizes between 100-500 nm. A comparison of hybridization on aldehyde-activated agarose-coated microarray slides and commercial amino-reactive microarray slides showed that aldehyde-activated agarose-coated slides had the highest signal-to-noise ratio of 850, suggesting that the aldehyde-activated agarose microarray slides are suitable in applications where analytes have a wide concentration range. By immobilizing the DNA probes using ultraviolet (UV) light, the signal-to-noise ratio was further increased to 3000 on the agarose microarray slides. The specificity of the UV cross-linked DNA probes was demonstrated using 21 and 25 bp long capture probes, enabling discrimination of target molecules differing in only one base. PMID- 15335223 TI - Elimination of background in film-based applications. AB - Erase-It Background Eliminator is a solution used directly on processed film to remove background or improve data resolution. Traditional methods, such as optimization of the scientific protocol or better estimation of exposure time, are tedious and uncertain. Nevertheless, autoradiography continues to be a simple, effective method to visualize data. Therefore, we evaluated the ability of Erase-It Working Solution to help solve background and resolution issues. To demonstrate the efficiency of the Background Eliminator, we analyzed the product's ability to remove signal evenly, performance on several brands of film, and usefulness with various detection methods. Even reduction of signal was demonstrated by performing densitometric analysis on film generated from a dot blot with serial dilutions of analyte. In addition, overexposed films from various suppliers were effectively treated to remove background and visualize data. Autoradiographs, generated with 32P-labeled probes, and chemiluminescent substrate were also processed resulting in clearer images. Our results demonstrate that film data can be treated quickly and conveniently without fear of artificial enhancement. We show the Background Eliminator to be a universal and timesaving tool to visualize results that otherwise may be difficult to interpret. PMID- 15335224 TI - New treatments for agitation. AB - Acute agitation is a frequent reason for emergency psychiatric intervention. It is important to intervene early to avoid escalation of agitation to aggression. Reducing risk by using effective treatments will result in fewer instances of seclusion and restraint, and fewer injuries to staff and patients. This paper will first review the epidemiology of aggressive behavior and mental disorders, followed by a discussion of assessment and diagnostic considerations. The pathophysiology of safety risk is discussed within the context of the model of the "triune brain." Pharmacological treatment strategies for acute episodes of agitated behavior will be discussed in detail. This includes newer formulations of novel antipsychotics such as liquids and rapidly disintegrating tablets, as well as intramuscular preparations. PMID- 15335225 TI - Is schizoaffective disorder a stable diagnostic category: a retrospective examination. AB - Debate continues about whether clear nosologic boundaries can be drawn between schizoaffective disorder (SA), schizophrenia (SP), and bipolar disorder (BPD). This study attempted to clarify these boundaries. A retrospective review of the records of adult psychiatric inpatients with DSM-IV diagnoses of SA (n = 96), SP (n = 245), and BPD (n = 203) was conducted. Patients were assessed at admission and discharge using standardized rating scales (completed by physicians and nurses) and self-report inventories. Differential improvement over time also was examined. Significant differences were found for gender, legal status at admission, age, LOS, episode number, and ethnicity. Overall, SA was rated by clinicians as intermediate between SP and BPD, although SA rated themselves as the most severe. SA was similar to SP on positive symptoms, intermediate on negative symptoms, and similar to BPD on mood- and distress-related symptoms. Independent of diagnosis, differences in change scores from admission to discharge were related to severity level at admission. Although several differences were found in symptom severity across domains, no syndrome was identifiable associated with the diagnosis of SA and the diagnosis was unstable over time, thereby bringing into question the validity of SA as a diagnostic entity. PMID- 15335226 TI - Special section: the therapeutic community in the 21st century. Introduction. PMID- 15335227 TI - The evolution of the therapeutic community. AB - The creation of Therapeutic Communities in the Military Hospitals of the UK for the treatment of psychological casualties of war during the Second World War is described. The personnel and the methods utilised are discussed and the lessons learned are summarised. It is argued that this was not an entirely new phenomenon. The author describes previous usage of similar methods in residential communities for disadvantaged or delinquent youth in the period between the two world wars and, prior to this, the eighteenth century "era of moral treatment" in mental hospitals in the UK and USA. PMID- 15335228 TI - The art and challenges of long-term and short-term democratic therapeutic communities. AB - This paper, cowritten by Kingsley Norton, since 1989 Director of Henderson Hospital (a therapeutic community founded by Maxwell Jones in 1947 in the United Kingdom), and Sandra Bloom, Founder of the Sanctuary Model in the United States, compares and contrasts the practice of the democratic therapeutic community (TC) as applied to the notion of long-term care (up to twelve months), to that of the democratic therapeutic milieu (TM) as applied to short-term care (up to one month). PMID- 15335229 TI - Democratic development of standards: the community of communities--a quality network of therapeutic communities. AB - As the inevitability of regulation and accountability dawned on the British Therapeutic Community movement at the end of the 1990s, a polarised debate took place. The product of that debate is now an action research based system of audit, with its principles and methods based on therapeutic community practice. This paper is written four years after the discussions started, and describes how the "Community of Communities" was conceived, what its methods are, some of the results from it first year of operation, and reflection about the nature of the process itself PMID- 15335230 TI - A culture of enquiry: research evidence and the therapeutic community. AB - This paper presents data from a systematic review and meta-analysis of 29 published studies of therapeutic community effectiveness using controls, including 8 randomised control trials. Meta-regressions suggest that the two types of therapeutic community, democratic and concept-based, and the age of the study, are the key sources of heterogeneity in the collection of studies analysed. Otherwise, heterogeneity is low and the meta-analysis confirms the effectiveness of therapeutic community treatment with overall summary log odds ratio for the 29 studies of 0.512 (95% ci -0.598 to -0.426). PMID- 15335231 TI - The therapeutic community as an adaptable treatment modality across different settings. AB - Simple core statements of the therapeutic community as a treatment modality are given, including a "living-learning situation" and "culture of enquiry." Applications are described in work with children and adolescents, chronic and acute psychoses, offenders, and learning disabilities. In each area the evolution of different therapeutic community models is outlined. In work with young people the work of Homer Lane and David Wills is highlighted. For long term psychosis services, the early influence of "moral treatment" is linked to the revitalisation of asylums and the creation of community based facilities; acute psychosis services have been have been run as therapeutic communities in both hospital wards and as alternatives to hospitalisation. Applications in prison are illustrated through an account of Grendon prison. The paper also outlines the geographical spread of therapeutic communities across many countries. PMID- 15335232 TI - [Mental health and psychiatric nursing. Various contributions to its history in Portugal]. PMID- 15335233 TI - [Aging and quality of life. Comparative study of elderly people living in the community and elderly people institutionalized in the region of Lisbon]. PMID- 15335234 TI - [Sexual education in adolescence (emotions and feelings)-- role of the nurse]. PMID- 15335235 TI - [Euthanasia-- live until the end. The Netherlands: a clarifying experience-- the failure of "controlled" euthanasia]. PMID- 15335236 TI - [Breastfeeding in the light of health psychology]. PMID- 15335237 TI - [Death in the hallways of the emergency department]. PMID- 15335238 TI - [Surviving the hospital-home transition]. PMID- 15335239 TI - [Diet and drug treatment of acute infant diarrhea]. PMID- 15335240 TI - Protective eyewear promotion: applying principles of behaviour change in the design of a squash injury prevention programme. AB - Eye injuries in squash have the potential to be severe. Although these injuries can be prevented through the use of protective eyewear, few players wear such eyewear. The aim of this paper is to outline the behavioural principles guiding the design of a squash eyewear promotion initiative, the Protective Eyewear Promotion (PEP). Ecological principles of behaviour change were used to provide a comprehensive perspective on intrapersonal factors, policies and physical environmental influences of protective eyewear use. Results of baseline player surveys and venue manager interviews were used to provide relevant and specific intervention content. At baseline, protective eyewear was not found to be readily available, and players' behaviours, knowledge and attitudes did not favour its use. The main components of PEP involved informing and educating both players and squash venue operators of the risk of eye injury and of appropriate protective eyewear, as well as assisting with the availability of the eyewear and offering incentives for players to use it. A structural strength of PEP was the strong collaborative links with the researchers of different disciplines, the squash governing body, eyewear manufacturers, squash venue personnel, as well as players. Attempts were made within the project structure to make provision for the future dissemination and sustainability of more widespread eye injury prevention measures in the sport of squash. PMID- 15335241 TI - Physical activity for people with a disability: a conceptual model. AB - The promotion of a physically active lifestyle has become an important issue in health policy in first-world countries. A physically active lifestyle is accompanied by several fitness and health benefits. Individuals with a disability can particularly benefit from an active lifestyle: not only does it reduce the risk for secondary health problems, but all levels of functioning can be influenced positively. The objective of this article is to propose a conceptual model that describes the relationships between physical activity behaviour, its determinants and functioning of people with a disability. The literature was systematically searched for articles considering physical activity and disability, and models relating both topics were looked for in particular. No models were found relating physical activity behaviour, its determinants and functioning in people with a disability. Consequently, a new model, the Physical Activity for people with a Disability (PAD) model, was constructed based on existing models of disability and models of determinants of physical activity behaviour. The starting point was the new WHO Model of Functioning and Disability, part of the International Classification of Functioning, Disability and Health (ICF), which describes the multidimensional aspects of functioning and disability. Physical activity behaviour and its determinants were integrated into the ICF model. The factors determining physical activity were based mainly on those used in the Attitude, Social influence and self-Efficacy (ASE) model. The proposed model can be used as a theoretical framework for future interventions and research on physical activity promotion in the population of people with a disability. The model currently forms the theoretical basis for a large physical activity promotion trial in ten Dutch rehabilitation centres. PMID- 15335242 TI - The dancer as a performing athlete: physiological considerations. AB - The physical demands placed on dancers from current choreography and performance schedules make their physiology and fitness just as important as skill development. However, even at the height of their professional careers, dancers' aerobic power, muscular strength, muscular balance, bone and joint integrity are the 'Achilles heels' of the dance-only selection and training system. This partly reflects the unfounded view, shared by sections of the dance world, that any exercise training that is not directly related to dance would diminish dancers' aesthetic appearances. Given that performing dance itself elicits only limited stimuli for positive fitness adaptations, it is not surprising that professional dancers often demonstrate values similar to those obtained from healthy sedentary individuals of comparable age in key fitness-related parameters. In contrast, recent data on male and female dancers revealed that supplementary exercise training can lead to improvements of such fitness parameters and reduce incidents of dance injuries, without interfering with key artistic and aesthetic requirements. It seems, however, that strict selection and training regimens have succeeded in transforming dance to an activity practised by individuals who have selectively developed different flexibility characteristics compared with athletes. Bodyweight targets are normally met by low energy intakes, with female dance students and professional ballerinas reported to consume below 70% and 80% of the recommended daily allowance of energy intake, respectively, while the female athlete 'triad' of disordered eating, amenorrhoea and osteoporosis is now well recognised and is seen just as commonly in dancers. An awareness of these factors will assist dancers and their teachers to improve training techniques, to employ effective injury prevention strategies and to determine better physical conditioning. However, any change in the traditional training regimes must be approached cautiously to ensure that the aesthetic content of the dance is not affected by new training techniques. Since physiological aspects of performing dance have been viewed primarily in the context of ballet, further scientific research on all forms of dance is required. PMID- 15335243 TI - The role of resistance exercise intensity on muscle fibre adaptations. AB - Although many training variables contribute to the performance, cellular and molecular adaptations to resistance exercise, relative intensity (% 1 repetition maximum [%1RM]) appears to be an important factor. This review summarises and analyses data from numerous resistance exercise training studies that have monitored percentage fibre type, fibre type cross-sectional areas, percentage cross-sectional areas, and myosin heavy chain (MHC) isoform expression. In general, relative intensity appears to account for 18-35% of the variance for the hypertrophy response to resistance exercise. On the other hand, fibre type and MHC transitions were not related to the relative intensity used for training. When competitive lifters were compared, those typically utilising the heaviest loads (> or =90% 1RM), that is weightlifters and powerlifters, exhibited a preferential hypertrophy of type II fibres when compared with body builders who appear to equally hypertrophy both type I and type II fibres. These data suggest that maximal hypertrophy occurs with loads from 80-95% 1RM. PMID- 15335245 TI - Pathologic quiz case. A 34-year-old woman with abdominal pain and a large right multicystic ovarian mass. Mullerian adenosarcoma arising in an endometriotic cyst. PMID- 15335244 TI - Factors associated with recurrent hamstring injuries. AB - A history of muscle injury represents a predominant risk factor for future insult in that muscle group. The high frequency of re-injury and persistent complaints after a hamstring strain comprise major difficulties for the athlete on return to athletic activities. Some of the risk factors associated with the possible recurrence of the injury are, in all probability, already implicated in the initial injury. One can distinguish between those events peculiar to the sport activity modalities (extrinsic factors) and other contributing factors based on the athletes individual features (intrinsic factors). For both categories, the persistence of mistakes or abnormalities in action represent an irrefutable component contributing to the re-injury cycle. Additional factors leading to chronicity can come from the first injury per se through modifications in the muscle tissue and possible adaptive changes in biomechanics and motor patterns of sporting movements. We emphasise the role of questionable approaches to the diagnosis process, drug treatment or rehabilitation design. To date, the risk factors examined in the literature have either been scientifically associated with injury and/or speculated to be associated with injury. In this context, quantifying the real role of each factor remains hypothetical, the most likely ones corresponding to inadequate warm-up, invalid structure and the content of training, muscle tightness and/or weakness, agonist/antagonist imbalances, underestimation of an extensive injury, use of inappropriate drugs, presence of an extensive scar tissue and, above all, incomplete or aggressive rehabilitation. Such a list highlights the unavoidable necessity of developing valid assessment methods, the use of specific measurement tools and more rigorous guidelines in the treatment and rehabilitation. This also implies a scientific understanding as well as specifically qualified medical doctors, physiotherapists and trainers acting in partnership. PMID- 15335246 TI - Pathologic quiz case. A 73-year-old man with a bladder mass. Combined primary small cell neuroendocrine carcinoma and high-grade papillary urothelial carcinoma of the urinary bladder. PMID- 15335247 TI - Nonproliferative fibrocystic changes and the Easter bunny. PMID- 15335248 TI - Epidermoid metaplasia of the cervix. PMID- 15335249 TI - Tracheal stenosis due to cytomegalovirus. PMID- 15335250 TI - One patient, two lymphomas. Simultaneous primary gastric marginal zone lymphoma and primary duodenal follicular lymphoma. AB - We report the case of a 73-year-old man who presented with a 2- to 3-month history of epigastric discomfort and guaiac-positive stool. An upper gastrointestinal endoscopy revealed a diffuse erythematous nodular mucosa and submucosal thickening in the stomach. Diffuse mucosal nodularity was also found in the second portion of the duodenum. A complete workup with histologic, immunohistochemical, and molecular studies revealed 2 distinct, apparently unrelated lymphomas, namely, a gastric marginal zone B-cell lymphoma (mucosa associated lymphoid tissue type) in a background of Helicobacter pylori gastritis and a grade 1/3 duodenal follicular lymphoma. The patient was then treated with an H pylori eradication regimen. No therapy was given for his duodenal follicular lymphoma because his symptoms were thought to be due to the gastric disease and because the duodenal lesion was small. A 6-month follow-up with upper gastrointestinal endoscopy revealed only focal biopsy scarring in the stomach and an apparently normal duodenum. The follow-up biopsies revealed significant regression of his mucosa-associated lymphoid tissue lymphoma, but persistence of his duodenal follicular lymphoma. The combination of these 2 lymphomas in the same patient and the different clinical responses to antibiotic treatment make this case unique. PMID- 15335251 TI - Nodular histiocytic hyperplasia of the endometrium. AB - A case of nodular histiocytic hyperplasia of the endometrium is described. A 45 year-old Japanese woman was found to have an enlarged uterus during her annual checkup. Hysterectomy and bilateral salpingo-oophorectomy specimens revealed uterine leiomyomas, adenomyosis, and acute salpingitis. A 5-mm, well-demarcated, elevated endometrial nodule (an incidental finding) was present and consisted of round or polygonal histiocytic cells with eccentric nuclei and pale or granular cytoplasm. The nuclei were ovoid, reniform, or crescent-shaped and had fine chromatin and inconspicuous nucleoli, and the cytoplasm contained single or multiple vacuoles. Immunohistochemically, the histiocytic cells were positive for vimentin, CD68, and lysozyme and were negative for cytokeratin, S100 protein, estrogen and progesterone receptors, and CD10. Nodular histiocytic hyperplasia in the endometrium is considered to be a reactive process. Differentiation from neoplasms, including signet-ring cell carcinoma, in curettage specimens is critical to avoid unnecessary surgical resection. PMID- 15335252 TI - Diagnosing allergic diseases in children. Practical recommendations for consulting pathologists. AB - In the environment of managed care, children with allergic diseases are increasingly likely to be evaluated by nonallergist physicians. While the presumptive diagnosis of an allergic disease can often be suspected on clinical grounds, signs and symptoms are not reliable for establishing a definitive diagnosis or for deciding on management options. This article discusses the use of immunoglobulin E antibody tests to classify children with allergic symptoms. The information is intended for pathologists who offer consultative laboratory services to nonallergist physicians. Emphasis is placed on defining the minimum number of tests necessary to identify allergic children at different ages. PMID- 15335253 TI - Pneumocystis carinii infection. Update and review. AB - OBJECTIVE: To review and update the literature on current trends with regard to Pneumocystis carinii (jiroveci ) diagnosis, treatment modalities, and its role in human disease processes. DATA SOURCES: Bibliographic databases (PubMed and Ovid) were searched for material and data between 1980 and September 2003 relevant to the review. Indexing terms used were "Pneumocystis carinii pneumonia," and "Pneumocystis jiroveci," with the English language as a constraint. Other sources were the PhD thesis of one of the authors (J.F.W., London University, 1993) and the library at the Arabian Gulf University in the Kingdom of Bahrain. STUDY SELECTION: Acquired immunodeficiency syndrome and organ transplant cases with Pneumocystis carinii pneumonia. DATA EXTRACTION: Independent extraction by 2 observers. DATA SYNTHESIS: We reviewed the major characteristics of P carinii (jiroveci ) with special emphasis on the more recently acquired data including the presence of a round pore in the cyst wall, which appears to be used for the release of sporozoites, supporting the hypothesis of sexual reproduction in P carinii (jiroveci ). CONCLUSIONS: Opportunistic infection with P carinii (jiroveci ) remains a significant cause of morbidity and mortality in human immunodeficiency virus and non-human immunodeficiency virus-associated immunosuppressed patients. Diagnosis may be achieved in the majority of cases by routine cytochemical stains and specialized techniques such as immunocytochemistry and polymerase chain reaction. The incidence of P carinii pneumonia can significantly be reduced with effective use of prophylaxis and early detection of cases at high risk. Immunization for P carinii pneumonia is in the early stages and presents a challenging area for research. PMID- 15335254 TI - Applications of flow cytometry and immunohistochemistry to diagnostic hematopathology. AB - OBJECTIVE: Diagnostic hematopathology depends on the applications of flow cytometric immunophenotyping and immunohistochemical immunophenotyping combined with the cytomorphology and histologic features of each case. Select cases may require additional ancillary cytogenetic and molecular studies for diagnosis. The purpose of this review is to focus on the applications of flow cytometric and immunohistochemical immunophenotyping of paraffin-embedded tissue to diagnostic hematopathology. Advantages and disadvantages of these techniques are examined. DATA SOURCES: The literature is extensively reviewed (PubMed 1985-2003) with an emphasis on the most recent applications and those that are most useful clinically, both diagnostically and prognostically. Study Selection.-Studies were selected based on statistically significant results in large studies with reported adequate clinical follow-up. DATA EXTRACTION: The methodology was reviewed in the selected studies to ensure reliable comparison of reported data. DATA SYNTHESIS: Flow cytometric immunophenotyping offers the sensitive detection of antigens for which antibodies may not be available for paraffin immunohistochemical immunophenotyping. However, paraffin immunohistochemical immunophenotyping offers preservation of architecture and evaluation of expression of some proteins, which may not be available by flow cytometric immunophenotyping. These techniques should be used as complimentary tools in diagnostic hematopathology. CONCLUSIONS: There are extensive applications of flow cytometric and immunohistochemical immunophenotyping to diagnostic hematopathology. As cytogenetic and molecular findings evolve in diagnostic hematopathology, there may be additional applications of flow cytometric and immunohistochemical immunophenotyping to this field of pathology. PMID- 15335255 TI - Routine syndecan-1 immunohistochemistry aids in the diagnosis of chronic endometritis. AB - CONTEXT: Chronic endometritis is reportedly observed in 3% to 10% of women undergoing endometrial biopsy for abnormal uterine bleeding. The diagnosis of chronic endometritis rests on the identification of the plasma cells. Their identification may be obscured by a mononuclear cell infiltrate, plasmacytoid stromal cells, abundant stromal mitoses, a pronounced predecidual reaction in late secretory endometrium, menstrual features, or secondary changes due to exogenous progesterone treatment prior to the biopsy. Syndecan-1 is a proteoglycan that is found on the cell surface of plasma cells and keratinocytes. Immunohistochemistry stains for this antibody may facilitate diagnosis of chronic endometritis. OBJECTIVE: To determine whether or not routine syndecan-1 immunohistochemistry will aid in the diagnosis of chronic endometritis. DESIGN: Immunohistochemistry stains for syndecan-1 were performed on 3 levels of 47 endometrial biopsies from patients with abnormal uterine bleeding. None of the patients had endometrial hyperplasia or an underlying malignancy. Clinical correlation and follow-up was attempted in 20 cases that showed evidence of plasma cells by syndecan-1 by immunohistochemistry. RESULTS: Plasma cells were identified in 20 cases, 7 of which were initially diagnosed as chronic endometritis. The remaining 13 positive cases were diagnosed as tubal metaplasia (1), secretory endometrium (4), proliferative endometrium (4), menstrual endometrium (1), endometrial polyp (1), secretory endometrium with endometrial polyp (1), and endometrial polyp with exogenous hormone effect (1) based on the original hematoxylin-eosin section. CONCLUSIONS: Syndecan-1 may be a useful adjunct in the diagnosis of chronic endometritis. Approximately half of the cases of chronic endometritis responded to an antibiotic regime; thus, this diagnosis is important and may potentially obviate the need for surgical intervention. PMID- 15335256 TI - President's address 2004: "wearing two hats": morphed perspectives on family quality of life. PMID- 15335257 TI - Believing is seeing: visual conventions in Barr's classification of the "feeble minded". AB - The eugenics era (c. 1900-1930) produced a strong desire among mental retardation professionals to recognize and control "the feeble-minded." Some eugenicists believed it was possible to classify individuals visually by learning to recognize what they believed to be observable characteristics of idiocy and imbecility. In this paper I used qualitative methods to examine the visual classification scheme developed by Martin Barr and conclude that most of this classification scheme is a social construction that was reliant on modes of presentation, such as clothing and photographic style, which had very little to do with any inherent feeble-mindedness. PMID- 15335258 TI - State of the States in developmental disabilities: 2004. AB - Public spending for MR/DD services grew rapidly during FYs 2000-2002. This rapid growth was followed by reductions in spending for MR/DD services as the nation's economy declined during 2002-2004. However, convergent factors stimulating future expansion of funding and services for persons with MR/DD include rapidly expanding cohorts of aging caregivers in the states, increasing longevity of persons with MR/DD, and extensive litigation in the states promoting access to services. PMID- 15335259 TI - Consumer-based quality of life assessment: the Maryland ask me! Project. AB - The concept of quality of life currently impacts program development, service delivery, management strategies, and outcome evaluation in the area of intellectual disabilities. Maryland uses peer interviewers to assess consumer perceived quality of life among adult recipients of MR/DD services and supports. In this article we describe the survey instrument and procedures and discuss assessment issues of quality of responses, acquiescence, and proxy respondents. We present the psychometric properties for eight core quality of life domains among 923 people assessed in FY 2001. Results are summarized and development of a model for enhancing social inclusion, personal development, and self determination was described. Service and personal characteristics relating to quality of life as well as some ways the results can be used for program enhancement are discussed. PMID- 15335260 TI - Training needs of direct support staff. AB - Training needs and current status of training for direct support staff were identified in one southeastern state. Data sources were (a) direct support staff, (b) administrators of direct support staff, and (c) consumers with developmental disabilities. Stratified sampling was used to gather data from 108 administrators and 170 direct support staff utilizing mail surveys and from 179 focus group participants (56 administrators, 53 direct support staff, and 70 consumers). Findings suggest that direct support staff recognized the needs (a) for additional training on Community Support Skill Standards; (b) to develop a comprehensive, cost-efficient statewide system for training direct support staff; and (c) to develop the role of direct support staff into a valued profession. PMID- 15335261 TI - Pathologic quiz case. Progressive headaches in a 50-year-old woman. Chordoid meningioma. PMID- 15335262 TI - Pathologic quiz case. A symptomatic renal tumor. Juxtaglomerular cell tumor. PMID- 15335263 TI - Pathologic quiz case. Progressive fatal encephalopathy in an immunosuppressed patient with a history of discoid lupus erythematosus. Subacute granulomatous meningoencephalitis (Acanthamoeba culbertsoni). PMID- 15335264 TI - Minimal (< or =0.1 cm) invasive carcinoma in breast core needle biopsies. Incidence, sampling, associated findings, and follow-up. AB - CONTEXT: Although minimally invasive (microinvasive) carcinoma (< or =0.1 cm) of the breast is a well-known and well-characterized entity in excision specimens, the significance of small foci of invasion in breast core needle biopsies has not been well described. OBJECTIVE: To define the significance of minimally invasive carcinoma in breast core needle biopsies. DESIGN: Review of a large series of core needle biopsies for invasive carcinomas measuring 0.1 cm or less and correlation of the results with those of subsequent excision. SETTING: Large community hospital. RESULTS: From approximately 8500 biopsies, a total of 18 cases of minimally invasive carcinoma from 16 women aged 42 to 80 years were identified. All were present on only 1 of 8 slides made from the block. Overall, the incidence was approximately 0.1% of all biopsies and 1% of all invasive carcinomas. Six cases were invasive lobular carcinomas, 1 was tubulolobular carcinoma, 3 were tubular carcinomas, and the remaining 8 were ductal carcinomas. Eight cases were associated with high-grade comedo ductal carcinomas, 2 with low grade ductal carcinoma in situ, 3 with atypical ductal hyperplasia, 3 with atypical ductal hyperplasia and lobular carcinoma in situ, and 2 with no other lesion. From a total of 8 sections done entirely through the block, the lesion was present on the first level in 4 cases and the fifth level in 5 cases. No cases were identified in the last 3 levels. Subsequent pathology was available for 16 of the 18 cases. Invasive carcinomas measuring more than 1 cm were present in 9 cases (64%; along with 2 positive lymph nodes), invasive carcinomas less than 1 cm in 2 cases (14%), ductal carcinoma alone in 4 cases (29%), and no carcinoma in 1 case (7%). No pathologic or radiologic features were associated with the finding of invasive carcinoma at excision. CONCLUSION: Invasive carcinoma measuring 0.1 cm or less is a rare finding in breast core needle biopsies, is commonly associated with in situ carcinomas and atypical hyperplasias, and is often associated with larger invasive foci at excision. However, invasive carcinomas smaller than 0.1 cm can occur without any other significant findings and may require relatively extensive sampling to identify. PMID- 15335265 TI - Febrile nonhemolytic transfusion reactions. Management by premedication and cost implications in adult patients. AB - CONTEXT: Febrile nonhemolytic transfusion reactions (FNHTRs) cause unwelcome interruptions during the course of blood product transfusions and necessitate measures to verify the nature of the reaction and to exclude certain dangerous reactions, such as hemolytic and septic phenomena. OBJECTIVE: To examine transfusion medicine data to determine the clinical implications of the routine administration of antipyretic medication to adult patients before transfusion for the prevention of FNHTRs. DESIGN: A retrospective review was conducted of FNHTR data during 5 years (1998-2002), and a determination was made of the cost of a transfusion complicated by an FNHTR. In addition, a comparative cost analysis was performed using our data and published data on the incidence of FNHTRs. The clinical implications of medication with respect to possible drug-induced adverse effects were assessed, as well as the potential interference with diagnosing other forms of transfusion reactions and the mitigation of the clinical effect of an FNHTR. RESULTS: For nearly 120,000 U of transfused blood components, approximately 80% of which were preceded by antipyretic medication during the study period, the overall incidence of FNHTR was found to be 0.09%. Furthermore, there was no evidence of antipyretic-associated complications, nor any evidence that antipyretics prevented the recognition of other more dangerous complications of transfusions. CONCLUSION: Our findings indicate that this practice provides significant advantages to the recipient of a transfusion, but does not appear to yield significant cost benefits for the health care provider. PMID- 15335266 TI - Chondroid syringoma. Cytokeratin 20 immunolocalization of Merkel cells and reappraisal of apocrine folliculo-sebaceous differentiation. AB - CONTEXT: Chondroid syringoma (CS) is a benign cutaneous adnexal tumor with epithelial and stromal components. Epithelial components derived from folliculo sebaceous-apocrine germ are evident in apocrine but not in eccrine CS. OBJECTIVES: To further characterize pilosebaceous differentiation and to identify the presence of Merkel cells in the areas of follicular differentiation. DESIGN: Histologic type, folliculo-sebaceous differentiation, character of stroma, and presence or absence of Merkel cells by cytokeratin (CK) 20 immunoreactivity were evaluated in 25 CSs (22 apocrine and 3 eccrine) from the surgical pathology files of Henry Ford Hospital (Detroit, Mich). RESULTS: Most CSs occurred in the head and neck region of patients aged 40 years or older. We found no significant difference in sex, age, or location between apocrine and eccrine types. The stroma varied from myxoid (100%) to chondroid (59%), with various amounts of fat (59%) and ossification identified in 2 cases (9%) of apocrine type, but was homogeneously myxoid in the eccrine type. Follicular and sebaceous differentiation was found in 64% and 32% of apocrine CSs, respectively. Only 2 (14%) apocrine CSs with follicular differentiation were positive for CK20 (a few scattered cells in one case and numerous grouped cells in the other in association with follicular epithelium). No correlation was found between type of stroma and the presence of Merkel cells. Scattered Merkel cells were identified in 83% of normal hair follicles and in 33.3% of normal epidermis. CONCLUSION: A high proportion of apocrine CSs show folliculo-sebaceous differentiation. The presence of Merkel cells in foci of follicular differentiation of CS supports the hypothesis that Merkel cells may be an integral constituent of follicles. To our knowledge, the presence of Merkel cells in CS, particularly in proliferative form, has not been described previously in the literature. PMID- 15335267 TI - Epidermal growth factor receptor status in breast cancer metastases to the central nervous system. Comparison with HER-2/neu status. AB - CONTEXT: The development of drug therapies (ZD1839) targeting epidermal growth factor receptor (EGFR) offers a pragmatic reason for exploring expression of EGFR in breast cancer, particularly metastatic breast cancer. There is a reported synergistic relationship between trastuzumab and ZD1839 therapy in patients with breast cancer. Although EGFR is the preferred dimerization partner for HER-2, it is unclear whether expression of these 2 interrelated receptors in a given patient with breast cancer would be parallel or mutually exclusive. OBJECTIVES: To assess EGFR status in primary breast carcinoma versus metastatic central nervous system (CNS) sites and to compare results with HER-2/neu status in the same tumor. DESIGN: Central nervous system metastases (n = 51) from 33 patients and corresponding primary breast cancer specimens, when available (n = 11), were immunohistochemically stained for EGFR using a monoclonal mouse anti-EGFR antibody (clone 31G7) that recognizes both the wild-type form and the 145-kd variant III form of EGFR. The sections were evaluated by visual and image analysis techniques, and results were compared to previously assessed HER-2/neu status. RESULTS: Epidermal growth factor receptor expression was found in CNS metastases from 39% of patients, with 82% concordance between the EGFR status of the primary breast and metastatic sites, and 92% concordance between the EGFR status among multiple CNS metastases in a given patient. Epidermal growth factor receptor and HER-2/neu status were concordant at the primary site in only 45% of patients. Additionally, EGFR and HER-2/neu status were concordant among multiple CNS metastases per individual case in only 45% of patients. CONCLUSION: Thirty nine percent of patients with metastatic breast cancer express EGFR, with parallel expression between metastatic sites and the primary neoplasm in 82% of the cases. The discordance in 18% of the cases, however, suggests that anti-EGFR agents might not show equal efficacy against metastatic tumor deposits and the primary tumor within a given patient. An additional corollary for pathologists based on this nonhomogeneity of receptor expression is that both the primary breast and multiple metastatic tumor deposits may need to be individually assessed for EGFR status. In our study, most metastatic tumor deposits showed expression for either EGFR or HER-2/neu, and less often for both, implying that drug therapies could be individualized for patients based on test results for both receptors. PMID- 15335270 TI - College of American Pathologists Laboratory Accreditation checklist item TRM.44955. Phase I requirement on bacterial detection in platelets. PMID- 15335268 TI - Distinct secreted Frizzled receptor protein 1 staining pattern in patients with hyperplastic polyposis coli syndrome. AB - CONTEXT: Patients with hyperplastic polyposis coli syndrome are thought to harbor precursor lesions of a proposed hyperplasia-carcinoma pathway in colorectal cancer, but morphologic recognition of such lesions remains difficult. Hypermethylation of the secreted Frizzled receptor protein 1 gene on chromosome 8p12 is one of the earliest molecular alterations in colorectal carcinogenesis, potentially disrupting the Wnt signaling cascade of cellular growth control. OBJECTIVE: To determine if hyperplastic polyps from patients with hyperplastic polyposis coli syndrome show a distinct immunohistochemical expression pattern for mismatch repair proteins and secreted Frizzled receptor protein 1 compared to their sporadic counterparts. DESIGN: Immunohistochemical studies (secreted Frizzled receptor protein 1, 3 mismatch repair proteins, and p53) were performed on 23 hyperplastic polyps, 6 synchronous colon cancers, and normal colonic mucosa from 6 patients with hyperplastic polyposis coli syndrome and were compared with studies of sporadic hyperplastic polyps obtained from 13 matched control subjects. RESULTS: The staining pattern for the mismatch repair proteins MLH-1, MSH-2, and MSH-6 did not differ between sporadic and syndromic hyperplastic polyps. In contrast, 52% of syndromic hyperplastic polyps showed a reproducible and distinct staining pattern for secreted Frizzled receptor protein 1 that was not seen in control specimens and that was associated with larger polyp size (P =.002) and location in the proximal colon (P =.01). CONCLUSIONS: Some hyperplastic polyps from patients with hyperplastic polyposis coli syndrome show a secreted Frizzled receptor protein 1 immunophenotype that could indicate alterations of cellular growth control. These findings may help identify precursor lesions in the proposed hyperplasia-carcinoma pathway of colorectal carcinogenesis. PMID- 15335272 TI - Pathologic quiz case. A 52-year-old woman with a pleural mass. Metastatic hemangiopericytoma. PMID- 15335273 TI - Tattoo pigments: popular fashion accessories or health hazards? PMID- 15335275 TI - UV-A fingerprint mutations in human skin cancer. AB - This review of our work, presented at the Photocarcinogenesis Symposium of the 14th International Congress on Photobiology, shows that UV-A causes a similar number of gene mutations as UV-B in human skin cancer. Areas of about 20 keratinocytes from solar keratoses and squamous cell carcinomas, which are benign and malignant skin cancers, respectively, were sampled by laser capture microdissection. Automated sequencing of the p53 gene was used to detect mutations in these tumor areas, and the cause of the mutations was attributed on the basis of previously published studies. UV-A and UV-B caused similar numbers of p53 gene mutations in both benign and malignant human skin tumors, with UV-B induced mutations being restricted to the upper areas of the tumors and UV-A induced mutations predominating at the basal layer. Furthermore, each microdissected region within a tumor had distinct mutations showing that the skin tumors consisted of different clones of cells. This is not consistent with how human skin carcinogenesis is currently understood, and hypotheses to explain our data are presented. We propose that the UV-A waveband of sunlight is as important as UV-B in causing skin cancer in humans. PMID- 15335279 TI - Psychotherapy for persons with mental retardation. PMID- 15335280 TI - The changing field of vaccine development in the genomics era. AB - An improved understanding of the human immune system and the genetic make-up of pathogens, together with advances in instrumentation and bioinformatics, have provided new insights into the variation of immune responses to vaccines within the human population. Pathogen variation and the diversity of the immune system components within the human population make the design of universal vaccines difficult. New subunit vaccines that target immunologically similar subgroups of the human population and representative pathogen variants are emerging from research that combines immunomics, pathogen genomics, and high-throughput instrumentation. PMID- 15335282 TI - Aligning the stakeholders. PMID- 15335284 TI - Using microarrays to predict resistance to chemotherapy in cancer patients. AB - Chemotherapy resistance remains a major obstacle to successful treatment and better outcome in cancer patients. The advent of whole genome experimental strategies, such as DNA microarrays, has transformed the way researchers approach cancer research. There is considerable hope that microarray technology will lead to the identification of new targets for therapeutic intervention, a better understanding of the disease process, and, ultimately, to higher survival rates and more personalized medicine. The question at hand is what is the best approach to apply these new technologies to the study of anticancer drug resistance, and how can the results obtained in the laboratory be quickly moved to a clinical setting? This review offers an overview of the microarray technology, including its recently associated strategies, such as array comparative genomic hybridization and promoter arrays. It also highlights some recent examples of microarray studies, which represent a first step toward a better understanding of drug resistance in cancer and, ultimately, personalized medicine. PMID- 15335285 TI - Pharmacogenomics and stomach cancer. AB - In subgroups of gastric cancer patients, chemotherapy treatments carry a high risk of toxicity without any clear evidence of antitumor activity. Individualization of therapy is required to treat each patient with the optimal drug and dose. Genetic polymorphisms are the hereditary determinants for interindividual variations of drug effect and the genetic approach represents a new tool to design a tailored therapy. This review focuses on the relevance of the host polymorphisms involved in metabolism, cellular transport and interaction with molecular targets of the drugs used in gastric cancer in conventional or innovative chemotherapy regimens. Pharmacogenetic studies based on a single gene or multi-gene approach (pharmacogenomics) are promising to identify gastric cancer patients at risk for adverse toxicity, but larger and controlled studies are needed to justify changes in the chemotherapeutic strategies. PMID- 15335286 TI - Pharmacogenetics of antiretroviral therapy: genetic variation of response and toxicity. AB - The application of a pharmacogenetic approach to antiretroviral drug therapy represents a significant challenge, as treatment involves multiple drugs and drug classes with the potential for significant variability in drug-host, as well as drug-drug, interactions. However, despite this inherent complexity, considerable gains have been made in understanding how genetic factors influence the efficacy and toxicity of HIV therapy. In this review the available evidence regarding genetic variation in drug disposition will be examined, including the potential for relatively polymorphic drug-metabolizing enzymes (e.g., cytochrome P450 isoforms) and drug transporters (e.g., P-glycoprotein) to influence the disposition of HIV protease inhibitor and non-nucleoside reverse transcriptase inhibitor drugs. In addition, the role of genetic variation in determining the immune response to drug-specific antigens will be considered as a potentially significant determinant of susceptibility to idiosyncratic drug reactions (e.g., major histocompatibility complex alleles associated with abacavir hypersensitivity). The current and potential clinical utility of pharmacogenetic testing in HIV management will also be emphasized. PMID- 15335287 TI - Postgenomic characterization of G-protein-coupled receptors. AB - G-protein-coupled receptors (GPCRs) constitute one of the largest families of membrane-spanning proteins. Their importance in drug development has been proven over and over again. Therefore, they remain one of the most significant groups of molecules to be characterized. In the postgenomic era, the methods used for the characterization of GPCRs have dramatically changed: the predicted orphan receptors are now often used to ascertain the ligands (reverse pharmacology), whereas, in the past, the bioactive ligand was used to identify the receptor (classic approach). In this review, we will give an overview of the recent postgenomic functional assays that are frequently used to link the orphan GPCR of both vertebrate and invertebrate organisms with their ligands. PMID- 15335288 TI - Toward individualized pharmaceutical care of East Asians: the value of genetic testing for polymorphisms in drug-metabolizing genes. AB - Research into the relationship between genetics and drug response has focused on polymorphisms in genes that encode drug-metabolizing enzymes, particularly the genes of cytochrome P450 superfamily 2, which affect the clearance of the anticoagulant warfarin, proton pump inhibitors, tricyclic antidepressants, and many other clinically relevant drugs. Much of this work has targeted East Asians, a genetically distinguishable and populous group. Researchers have identified polymorphisms that inactivate gene function, compared polymorphism frequencies in East-Asian and Caucasian populations, and determined the effects on the pharmacokinetic parameters of drugs. Detection in an individual of polymorphisms known to inactivate a drug-metabolizing enzyme is predictive of poor metabolism of drugs processed by that pathway, which itself may be predictive of an atypical drug response. Genetic tests can be used to screen for individuals with poor metabolizer phenotypes, with the ultimate goal of predicting the clinical effects of drugs. PMID- 15335289 TI - Pharmacogenetics of dopamine receptors and response to antipsychotic drugs in schizophrenia--an update. AB - A considerable number of pharmacogenetic studies have been performed in recent years to define the association of antipsychotic medication response with dopamine receptor polymorphisms and, despite contradictory results, decisive trends have emerged. For the dopamine D2 receptor (DRD2), a trend toward an association with favorable response seems to emerge for the -141C Ins allele of the DRD2 -141C Ins/Del polymorphism and the A1 allele of the Taq1A polymorphism. In the case of the D3 receptor, the Ser9Gly polymorphism has been extensively investigated and a pattern of association is seen between the Ser9 allele and a response to typical antipsychotics, and between the Gly9 allele and a response to atypical antipsychotics. For the D4 receptor, no convincing association results have been reported to date. These trends are discussed with regard to methodological directives and functional implications. PMID- 15335290 TI - DNA methylation biomarkers of cancer: moving toward clinical application. AB - While different markers for cancer diagnosis have been known for at least a decade, the systematic search for biomarkers emerged only several years ago. In this article, I will concentrate on DNA methylation as a dynamic and robust platform for the development of cancer-specific biomarkers. Simultaneous analysis of a growing number of independent methylation events can create increasingly more precise and individualized diagnostics. The differential detection of methylated and unmethylated DNA can be accomplished through either chemical modification or digestion with methylation-sensitive restriction enzyme(s). The benefits and potential pitfalls of both these approaches for clinical sample analysis will be addressed. PMID- 15335291 TI - Research issues and strategies for genomic and proteomic biomarker discovery and validation: a statistical perspective. AB - The development and validation of clinically useful biomarkers from high dimensional genomic and proteomic information pose great research challenges. Present bottlenecks include: that few of the biomarkers showing promise in initial discovery were found to warrant subsequent validation; and biomarker validation is expensive and time consuming. Biomarker evaluation should proceed in an orderly fashion to enhance rigor and efficiency. A molecular profiling approach, although promising, has a high chance of yielding biased results and overfitted models. Specimens from cohorts or intervention trials are essential to eliminate biases. The high cost for biomarker validation motivates some novel study design features, including sequential filtering and DNA pooling. For data analysis, logistic regression (in particular, boosting logistic regression) has features of robustness against model misspecification, and has resistance to model overfitting. Model assessment and cross-validation are critical components of data analysis. Having an independent test set is a vital feature of study design. PMID- 15335292 TI - Functional informatics: convergence and integration of automation and bioinformatics. AB - The biopharmaceutical industry is currently being presented with opportunities to improve research and business efficiency via automation and the integration of various systems. In the examples discussed, industrial high-throughput screening systems are integrated with functional tools and bioinformatics to facilitate target and biomarker identification and validation. These integrative functional approaches generate value-added opportunities by leveraging available automation and information technologies into new applications that are broadly applicable to different types of projects, and by improving the overall research and development and business efficiency via the integration of various systems. PMID- 15335293 TI - Report on the joint EFPIA, DIA and EMEA pharmacogenetics workshop: moving toward clinical application. PMID- 15335294 TI - Iconix Pharmaceuticals, Inc.--removing barriers to efficient drug discovery through chemogenomics. AB - Iconix Pharmaceuticals, Inc. is pioneering the new field of chemogenomics--the integration of chemistry and genomics to profile, optimize and monitor clinical candidates and drug treatment. Iconix's chemogenomic capabilities enable pharmaceutical companies to increase the odds of advancing the right compounds to the clinic and managing the development of those compounds in human testing, thus reducing attrition rates and the costs of drug discovery. PMID- 15335295 TI - Considerations with the use of biological therapy in the treatment of rheumatoid arthritis. AB - The treatment of rheumatoid arthritis (RA) has changed dramatically over the past 15 years with the realisation that earlier, aggressive therapy limits progression. There is evidence that biological response modifiers (BRMs), which target specific cytokines such as TNF-alpha and IL-1, are more effective than traditional disease-modifying antirheumatic drugs (DMARDs), especially in combination with methotrexate. Four therapies are approved for use in RA; three target TNF-alpha (etanercept [Enbrel, Amgen Inc.], infliximab [Remicade, Centocor Inc.], and adalimumab [Humira, Abbott]), and one targets IL-1 (anakinra [Kineret, Amgen Inc.]). It is clear from both the clinical trials and postmarketing reports that all four agents have a different safety profile compared with traditional DMARDs. There are several areas of concern with the use of the BRMs, which include serious and opportunistic infections, malignancy/lymphoma, congestive heart failure, demyelination, injection/infusion reactions, development of autoantibodies and lupus-like disease. It is important to be fully aware of the safety profile and differences between BRMs in order to use them appropriately. PMID- 15335296 TI - The safety and efficacy of linezolid in orthopaedic practice for the treatment of infection due to antibiotic-resistant organisms. AB - Linezolid is the first of a new class of antibacterial agents, the oxazolidinones. It is particularly effective against Gram-positive infections and little resistance has been reported, even amongst methicillin- and vancomycin resistant bacteria. The compound's excellent oral bioavailability and reasonable safety profile, along with the increasing incidence of resistant infections, means that linezolid offers a valuable alternative to more traditional therapies such as vancomycin. Although no large randomised trials have been carried out in patients with orthopaedic infections such as osteomyelitis and septic arthritis, early results are encouraging. However, the apparent increase in observed adverse events, particularly bone marrow suppression, seen with prolonged administration, means that treatment of such patients must be undertaken with careful surveillance, at least until these complications are better understood. PMID- 15335297 TI - Levetiracetam safety profiles and tolerability in epilepsy patients. AB - This review discusses the safety and tolerability of levetiracetam, as presented by the available literature, with attention paid to special populations. In Phase II/III trials, the adverse effects occurring more commonly in the treatment groups versus the placebo group were; somnolence (14.8 versus 8.4%), asthenia (14.7 versus 9.1%), infection (primarily common cold) (13.4 versus 7.5%), and dizziness (8.8 versus 4.1%). Adverse events usually appear within the first month after treatment initiation, are not dose-dependent, are mostly mild-to-moderate, generally resolve without medication withdrawal, and are transient when the medication is stopped. No significant changes in haematology and chemistry profiles or weight occurred. Hypersensitivity reactions were rare and no idiosyncratic event has been reported. Open-label studies have added patient data with other epileptic syndromes and from a wider patient pool, such as children and patients with prior psychiatric history. These studies have supported initial safety findings, but have reported increased behavioural adverse events in children and patients with a history of prior behavioural problems. Levetiracetam is proving to be safe and well-tolerated. So far, it appears to have a favourable safety profile in special populations, such as children, the elderly, and patients with hepatic dysfunction. Preliminary data in pregnancy are promising, but more data are needed on the impact of levetiracetam on the developing fetus and pharmacokinetic alterations caused in pregnancy. Adjustments in dosing are required for decreases in renal clearance. PMID- 15335298 TI - The benefits and risks associated with cholinesterase inhibitor therapy in Alzheimer's disease. AB - The 'second-generation' cholinesterase inhibitors (ChEIs), donepezil, galantamine and rivastigmine, are a class of medications that are currently approved for the treatment of mild-to-moderate Alzheimer's disease (AD). These medications have proven efficacy in improving cognition, behaviour, activities of daily living, and global functioning in mild-to-moderate AD. They have also been shown to reduce caregiver stress and to delay time to nursing home placement. Two separate meta-analyses have indicated that ChEIs confer a modest but significant therapeutic benefit in the treatment of AD, despite higher rates of treatment discontinuation and side effects than placebo. There is growing evidence to support their efficacy in treating moderate-to-severe AD. ChEIs are generally well-tolerated, with side effects that tend to be dose-related and are most problematic during dose titration. The most common adverse effects, related to cholinergic stimulation in the brain and peripheral tissues, include gastrointestinal, cardiorespiratory, extrapyramidal, genitourinary, and musculoskeletal symptoms, as well as sleep disturbances. Few clinically significant drug-drug interactions with ChEIs have been identified. Three head-to head trials of ChEIs in the treatment of AD have been published to date, but are limited due to their open-label design, rates of titration, and the drug dosage levels utilised. Further study is needed to examine other indications for ChEIs, as well as their combination with newer treatments, such as memantine. PMID- 15335299 TI - Cocaine-induced renal disease. AB - Cocaine has anaesthetic, vasoconstrictive and CNS stimulatory effects. Presently, it is used clinically as a local anaesthetic and abused as a recreational drug. It has been implicated in both acute and chronic renal failure and has been reported to affect every aspect of the nephron. This article will review the spectrum of cocaine-induced kidney disease and attempt to give insight into the pathophysiological mechanisms involved. PMID- 15335300 TI - Information technology for detecting medication errors and adverse drug events. AB - It is estimated that over three-quarters of a million people are injured or die in hospitals each year from adverse drug events (ADEs). The majority of medical errors result from poorly designed healthcare systems rather than from negligence on the part of healthcare providers. In general, healthcare systems rely on voluntary reporting, which seriously underestimates the number of medication errors and ADEs by as much as 90%. This paper reviews the causes and impact of medication errors and ADEs. It also reports studies that have used information technology (IT) to detect and prevent medication errors and ADEs. Significant reduction of medication errors and ADEs requires systemic implementation of IT, improvements in the reporting of errors, and integration of the components of the healthcare systems' information systems. At the present time, most healthcare systems should be able to use IT to detect and prevent ADEs. PMID- 15335301 TI - Sexual dysfunction associated with antidepressant drugs. AB - The term 'sexual dysfunction' describes a disturbance in sexual desire and the psychophysiological changes that characterise the normal sexual response cycle, and cause marked personal distress and interpersonal difficulty. Epidemiological studies indicate that sexual dysfunction is common in the general population, but more common in depressed individuals in community settings and clinical samples. Most antidepressant drugs have adverse effects on sexual function, but accurate identification of the incidence of treatment-emergent dysfunction has proved troublesome. Futhermore, investigations of sexual dysfunction associated with antidepressants have one or more methodological flaws. There may be some advantages for bupropion, moclobemide, nefazodone and reboxetine over other antidepressants. Many approaches have been adopted for management of patients with sexual dysfunction associated with antidepressant treatment, including waiting for the problem to resolve, behavioural strategies to modify sexual technique, individual and couple psychotherapy, delaying the intake of antidepressants until after sexual activity, reduction in daily dosage, 'drug holidays', use of adjuvant treatments, and switching to a different antidepressant. PMID- 15335302 TI - Drug-induced urticaria. AB - Urticaria is characterised by transient swellings of the skin, which fluctuate over hours. Deeper swellings of the subcutaneous and submucosal tissue are known as angio-oedema. Drug-induced urticaria has been reported with a wide range of drugs and vaccines. NSAIDs and antibiotics are the drugs most commonly associated with urticaria, although reliable data from prospectively controlled studies is scarce. Spontaneous reports of drug-induced urticaria to the Committee on Safety of Medicines, UK, over a 40-year period also implicate bupropion, selective serotonin re-uptake inhibitor antidepressants, angiotensin-converting enzyme inhibitors (ACEI), H2 and H1 antihistamines, and systemic antifungals. New evidence suggests that selective COX-2 inhibitors may be tolerated in patients with aspirin-sensitive urticaria. The safety of angiotensin II receptor antagonists in patients with angio-oedema induced by ACEI has not yet been established. PMID- 15335303 TI - Safety of benzodiazepines in the geriatric population. AB - Benzodiazepines are the most frequently prescribed antianxiety drugs in the elderly. Despite their usefulness and safety in the younger population, there is concern about the rationale for their use in the elderly. Studies of the therapeutic use of benzodiazepines in the elderly are rare. Elderly females with co-morbid medical and psychiatric conditions and who are taking multiple medications form the group most frequently prescribed benzodiazepines among the elderly, and the group most likely to experience side effects. Age-related pharmacokinetic and pharmacodynamic changes increase the potential for certain side effects in the elderly. Significant adverse effects that may be associated with benzodiazepine use in the elderly include falls, cognitive impairment, sedation, and impairment of driving skills, all of which are particularly related to the long half-life of benzodiazepines. Long-term use of benzodiazepines should be discouraged because of the risk of dependence, which is a serious problem in the elderly. Unrecognised and untreated benzodiazepine dependence can lead to serious medical complications. PMID- 15335304 TI - Safety and side effect profile of fluoxetine. AB - Fluoxetine was the first selective serotonin re-uptake inhibitor to be widely available for treatment of depression and numerous other neuropsychiatric disorders. Its attributes have been described in numerous scientific papers, and it has been the subject of a considerable volume of lay press. Fluoxetine is generally safe and well-tolerated. Common adverse events reported with the recommended dose of 20 mg/day are referable to the gastrointestinal system and the nervous system. The approved dose range is up to 80 mg/day, and when higher doses are used, adverse events are more common. The long half-life of fluoxetine and its active metabolite essentially preclude a withdrawal phenomenon. It is an inhibitor of cytochrome P450 (CYP) 2D6 and other CYP enzymes, which increases the potential for drug interactions. However, most of these are not clinically important. The purpose of this review is to provide an overview of some of the most important information related to safety and side effects of this drug. PMID- 15335305 TI - Gene transfer in tissue repair: status, challenges and future directions. AB - Wound repair involves a complex interaction of various cell types, extracellular matrix molecules and soluble mediators. Details on signals controlling wound cell activities are beginning to emerge. In recent years this knowledge has been applied to a number of therapeutic strategies in soft tissue repair. Key challenges include re-adjusting the adult repair process in order to augment diseased healing processes, and providing the basis for a regenerative rather than a reparative wound environment. In particular, the local delivery of pluripotent growth factor molecules to the injured tissue has been intensively investigated over the past decade. Limited success of clinical trials indicates that an important aspect of the growth factor wound-healing paradigm is the effective delivery of these polypeptides to the wound site. A molecular genetic approach in which genetically modified cells synthesise and deliver the desired growth factor in a time-regulated manner is a powerful means to overcome the limitations associated with the (topical) application of recombinant growth factor proteins. This article summarises repair mechanisms and their failure, and gives an overview of techniques and studies applied to gene transfer in tissue repair. It also provides perspectives on potential targets for gene transfer technology. PMID- 15335306 TI - Immunomodulation to combat atherosclerosis: the potential role of immune regulatory cells. AB - Atherosclerosis is an inflammatory disease of the arterial wall that carries an important socio-economic burden. The available data strongly suggest that both innate and adaptive immuno-inflammatory mechanisms are the major determinants of plaque complications (called instability). Therefore, most of the important advances in the comprehension of the mechanisms of atherosclerosis came from studies that aimed at the elucidation of the critical components involved in the modulation of the immuno-inflammatory response in experimental models of atherosclerosis. As the T helper (Th)1-driven immune response has been consistently shown to promote atherosclerosis, the general belief is that immunomodulation through Th2 may be suitable to halt the disease process. Here, a novel view of the immuno-inflammatory response in atherosclerosis is presented, in which the natural and/or adaptive regulatory responses modulate both Th1 and Th2 responses, and play a central role in counteracting disease initiation and progression. PMID- 15335307 TI - Ocular neovascularisation and excessive vascular permeability. AB - Diseases complicated by vascular leakage and/or neovascularisation in the eye are responsible for the vast majority of visual morbidity and blindness in developed countries. The molecular signals that control vascular permeability and neovascularisation in the eye are being defined. Members of the vascular endothelial growth factor (VEGF) family are key stimulators that interact with two tyrosine kinase receptors, VEGF receptor (VEGFR)1 and 2; binding to two other receptors that lack tyrosine kinase activity, the neuropilins, is also important. Signalling through the VEGF pathway is modulated by the Tie2 receptor and its binding partners, the angiopoietins. Each of the participants in these two signalling pathways provides a potential target for intervention. Several proteins with antiangiogenic activity balance the stimulators and the outcome is determined by the net balance. Endostatin suppresses vascular permeability as well as ocular neovascularisation, suggesting that vascular leakage may also be regulated by counteracting proteins. Gene transfer provides a useful way to influence these balances. Clinical trials are underway to test whether these mechanisms can be translated into new therapies. PMID- 15335308 TI - Light-directed gene delivery by photochemical internalisation. AB - This article reviews a novel technology, named photochemical internalisation (PCI), for light-directed delivery of transgenes. Most gene therapy vectors are taken into the cell by endocytosis and, hence, are located in the endocytic vesicles. Although viral vectors have developed the means to escape from these vesicles, poor endosomal release is one of the major obstacles for non-viral vectors. PCI is a technology that allows liberation of the entrapped vectors carrying a gene in response to illumination. The method is based on chemical compounds (photosensitisers) that localise specifically in the membranes of endocytic vesicles and, following activation by light, induce the rupture of the vesicular membranes. The released transgenes can further be transferred to the nucleus, transcribed and translated. As gene liberation depends on light, enhancement of gene expression is achieved only at illuminated regions. PCI substantially improves gene transfer in vitro not only with non-viral gene vectors, but, surprisingly, also with adenoviruses and adeno-associated viruses. This article will review the background for the PCI technology and its role for gene delivery using both non-viral and viral vectors. Some aspects of the potential of PCI for site-specific gene delivery in therapeutic situations will also be discussed. PMID- 15335309 TI - Gene delivery approaches to heart failure treatment. AB - Heart failure remains a leading cause of worldwide morbidity and mortality. Despite recent advances in treatment and our increasing knowledge of pathophysiology and the molecular derangements involved in the failing heart, our ability to affect the underlying cardiac disease processes is limited. In recent years, there has been considerable interest in myocardial gene transfer as both an investigational and potential therapeutic modality. Ultimately, the goal of any such strategy is to reprogramme failing cardiac myocytes and correct the aberrant molecular events causing heart failure. So far, viral vectors have been utilised with success more frequently than any other method of gene delivery in animal models. Studies in animal models and in failing human cardiomyocytes in culture targeting specific molecular pathways, including the beta-adrenergic receptor cascade and the myocyte intracellular calcium handling system, have shown encouraging results and offer hope that gene manipulation may provide novel adjunctive therapeutic modalities for human heart failure. PMID- 15335310 TI - Gene therapy for chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is a congenital immune deficiency that is a promising therapeutic target for gene replacement into haematopoietic stem cells (HSCs). CGD results from mutations in any one of four genes encoding subunits of the superoxide-generating NADPH oxidase of phagocytes. Life-threatening, recurrent bacterial and fungal infections, as well as inflammatory granulomas, are the hallmarks of the disease. NADPH oxidase activity can be reconstituted by retroviral- or lentiviral-mediated gene transfer to human CGD marrow in vitro and in xenograft transplant models. Gene transfer studies in knockout mouse models that resemble the human disease suggest that correction of oxidase activity in a minority of phagocytes will be of clinical benefit. Phase I clinical studies in unconditioned CGD patients showed transient expression of small numbers of gene corrected neutrophils. Areas of research at present include efforts to enhance gene transfer rates into repopulating HSCs using vectors that transduce quiescent cells, and to increase the engraftment of genetically corrected HSCs using non myeloablative conditioning and drug resistance genes for selection. PMID- 15335311 TI - Gene therapy of epidermolysis bullosa. AB - Easy access to the organ and identification of underlying mutations in epidermolysis bullosa (EB) facilitated the first cutaneous gene therapy experiments in vitro in the mid-1990s. The leading technology was transduction of the respective cDNA carried by a retroviral vector. Using this approach, the genotypic and phenotypic hallmark features of the recessive forms of junctional EB, which are caused by loss of function of the structural proteins laminin-5 or bullous pemphigoid antigen 2/type XVII collagen of the dermo-epidermal basement membrane zone, have been corrected in vitro and in vivo using xenograft mouse models. Recently, this approach has also been shown to be feasible for the large COL7A1 gene (mutated in dystrophic EB), applying PhiC31 integrase or lentiviral vectors. Neither of these approaches has made it into a successful Phase I study on EB patients. Therefore, alternative approaches to gene correction, including modulation of splicing, are being investigated for gene therapy in EB. PMID- 15335312 TI - Tumour-targeted chemotherapy with immunoconjugates of calicheamicin. AB - Antibody-targeted chemotherapy is a therapeutic strategy in cancer therapy that involves a monoclonal antibody specific for a tumour-associated antigen, covalently linked via a suitable linker to a potent cytotoxic agent. Tumour targeted delivery of a cytotoxic agent in the form of an immunoconjugate is expected to improve its antitumour activity and safety. Calicheamicin is a cytotoxic natural product isolated from Micromonospora echinospora that is at least 1000-fold more potent than conventional cytotoxic chemotherapeutics. Calicheamicin binds DNA in the minor groove and causes double-strand DNA breaks, leading to cell death. Immunoconjugates of calicheamicin targeted against tumour associated antigens exhibit tumour-specific cytotoxic effects and cause regression of established human tumour xenografts in nude mice. Gemtuzumab ozogamicin is the first clinically validated cytotoxic immunoconjugate in which a humanised anti-CD33 antibody is linked to a derivative of calicheamicin. Gemtuzumab ozogamicin is indicated for the treatment of elderly patients with relapsed acute myeloid leukaemia. A similar conjugate, inotuzumab ozogamicin, is being evaluated at present in Phase I clinical trials in patients with non Hodgkin's lymphoma. A number of tumour-targeted immunoconjugates of calicheamicin are being explored preclinically at present for their therapeutic applications. PMID- 15335313 TI - Monoclonal antibodies for brain tumour treatment. AB - Conventional treatment of brain tumours includes surgical, radiotherapeutic and chemotherapeutic modalities. Nonetheless, the outcome of patients with brain tumours, in particular glioblastoma, remains poor. Immunotherapy with armed or unarmed monoclonal antibodies targeting tumour-specific antigens has emerged in the last two decades as a novel potential adjuvant treatment for all types of neoplasia. Many challenges to its implementation as a safe and viable therapy for brain tumours still need to be addressed; nevertheless, results from ongoing Phase I/II clinical trials are encouraging, as disease stabilisation and patient survival prolongation have been observed. Advances in preclinical and clinical research indicate that treatment of brain tumours with monoclonal antibodies can be increasingly adjusted to the characteristics of the targeted tumour and its environment. This aspect relies on the careful selection of the target antigen and corresponding specific monoclonal antibody, and antibody format (size, class, affinity), conjugation to the appropriate toxin or radioactive isotope (half life, range), and proper compartmental administration. PMID- 15335314 TI - Allergy vaccines--new approaches to an old concept. AB - Allergy vaccination (AV) consists of injecting increasing amounts of offending allergens into sensitive patients with the intention of reducing their level of sensitivity to allergens. This form of therapy was first used over one hundred years ago and until recently had not changed in principle. The vaccines themselves are now far better characterised and standardised, according to new regulatory requirements. The therapy is believed to exert its effects by a combination of means: by the induction of blocking antibodies; a switch from a T helper (Th)2 to a more Th1 allergen-specific immune response; and induction of anergy, probably via the development of allergen-specific regulatory T cells. New allergen forms and formulations are being designed with these targets in mind. Allergoids (allergens chemically modified to reduce allergenicity, but to retain immunogenicity) are becoming employed more frequently. More modern depot forms, such as those containing tyrosine or calcium phosphate, are replacing aqueous extracts and older depot adjuvants such as alum. T cell-reactive peptides and recombinant allergens or their muteins are also being studied as replacements for whole extracts and have shown some potential. Immunomodulators, such as monophosphoryl lipid A (MPL), designed with defined targets in mind are now included in some vaccines and help to accelerate the process. All these measures have led to a reduction in the need for the traditional long injection schedules. The authors are very familiar in particular with the background to the use of MPL as an adjuvant. They have been personally involved in the development of this approach, which has led to a product being available for use on a regular basis in some European countries. Hence, this work is reported in considerable detail. Other similar immunomodulators, such as CpG motifs, are in development, while new targets, such as the Notch protein/receptor interaction, are exciting new developments that may eventually bear fruit. The excellent safety profile of the sublingual route of administration of allergy vaccines could lead to the wider use of AV, and locally active immunomodulators could make AV a therapy of choice for many more patients than at present. PMID- 15335315 TI - HIV-1 vaccines and co-infection. AB - Vaccines are an economically efficient means of controlling viral infections, and it is likely that a vaccine against HIV-1 will be the most effective way of controlling the global AIDS crisis. However, an effective vaccine has not yet been attainable and in developing countries co-infection with protozoa and other chronic diseases adds another level of complexity to the design of an HIV-1 vaccine. Helminthic and protozoan infections can result in a constant state of immune activation that is characterised by a dominant T helper (Th)2 type of cytokine profile. Such an immune profile is likely to have an adverse impact on the efficacy of an HIV-1 vaccine CD8 cellular immune response and the corresponding Th1 cytokines that are most likely to be important for clearing viral infections. PMID- 15335316 TI - Tuberculosis vaccine development: research, regulatory and clinical strategies. AB - In the past decade, while the global tuberculosis (TB) epidemic has continued to devastate mankind, considerable progress has nevertheless been made in the development of new and improved vaccines for this ancient disease. Recombinant bacillus Calmette-Guerin strains, DNA-based vaccines, live attenuated Mycobacterium tuberculosis vaccines and subunit vaccines formulated with novel adjuvants have shown promise in preclinical animal challenge models. Three of these vaccines are being evaluated at present in human clinical studies, and several other vaccine preparations are being targeted for clinical trials in the near future. Although the preclinical characterisation and testing of new TB vaccines has clearly led to exciting new findings, complex regulatory and clinical trial design issues remain as a challenge to TB vaccine development. This report reviews some of the exciting advances in TB research that have led to the development of new TB vaccines, and addresses the unique regulatory and clinical issues associated with the testing of novel anti-TB preparations in human populations. PMID- 15335317 TI - Recent advances in vaccines for leishmaniasis. AB - The observation that recovery from infection with Leishmania confers immunity to reinfection suggests that control of leishmaniasis by vaccination may be possible. However, there are no vaccines available at present to control any form of leishmaniasis, despite considerable efforts. Studies of the immunopathogenesis and mechanisms of protective immunity, mainly derived from animal models of experimental leishmaniasis, have defined a number of features that should be met by an effective vaccine. In addition, several antigens have been identified that may be potential vaccine candidates, and molecular biological techniques have made them available as recombinant proteins for second-generation vaccines. Furthermore, molecules present in the saliva of Leishmania-transmitting vectors have been demonstrated as valuable candidates for the development of anti Leishmania vaccines. This review concentrates on the most promising vaccine candidates and highlights new approaches for the development of vaccines. Finally, based on present knowledge, the future prospects for developing an effective vaccine against the different clinical forms of leishmaniasis are discussed. PMID- 15335318 TI - Treatment and prevention of enterococcal infections--alternative and experimental approaches. AB - Enterococci are one of the leading types of organisms isolated from infections of hospitalised patients and the third most common cause of nosocomial bloodstream infections. They contribute significantly to patient mortality and morbidity, as well as healthcare costs. The emergence of resistance against virtually all clinically available antibiotics and the ability to transfer these resistance determinants to other pathogens demonstrates the urgency for an improved understanding of enterococcal virulence mechanisms, and the development of alternative treatment and prevention options. This article reviews new antimicrobials, vaccine targets, bacteriophage therapy, as well as treatments targeting virulence factors and biofilm, for their potential to treat and/or prevent enterococcal infections. Although clinical isolates often cause serious infections, so-called 'non-pathogenic' strains are used as therapeutics in the form of probiotics. Understanding the differences between true pathogens and beneficial commensals may help to evaluate future treatment and prophylactic options. PMID- 15335319 TI - CpG DNA: immunomodulation and remodelling of the asthmatic airway. AB - Asthma is a disorder of increasing severity and prevalence. Present understanding of the pathogenesis of asthma emphasises its inflammatory nature. Unbridled inflammation appears to induce irreversible changes, collectively known as airway remodelling. CpG oligonucleotides are a class of compounds that have been developed from studies of prokaryotic DNA. Bacterial DNA, for example, contains sequence motifs based on the dinucleotides cytosine-guanine (CpG); these motifs are suppressed in mammalian DNA and induce (as part of the innate immune system) inflammation, characterised by the induction of T helper type 1 and regulatory responses. The pattern of inflammation promoted by CpG DNA tends to suppress the cytokine and cellular responses characteristic of asthma and atopic disorders. This has led to the investigation and development of CpG DNA as a novel therapeutic approach for the treatment and prevention of these disorders. In addition to suppressing acute and persistent airway inflammation, CpG DNA also reduces the development of subepithelial collagen deposition, goblet cell hyperplasia/metaplasia, and other aspects of airway remodelling. In this paper, the effects of CpG DNA on asthma inflammation and remodelling are reviewed. PMID- 15335320 TI - First International Symposium on Melanoma and Other Cutaneous Malignancies. AB - The First International Symposium on Melanoma and Other Cutaneous Malignancies, held in New York City on 23-25 April 2004, brought together researchers and clinicians from all over the world to discuss recent advances in the prevention, treatment and diagnosis of melanoma and other cutaneous malignancies. Discussion topics included primary and secondary prevention; advances in surgical therapy, including sentinel and elective lymph node dissection; the biology and pathogenesis of melanoma, including pathways of drug resistance; genomic analysis of melanoma, serum and tumour cell markers; with point and counterpoint sessions debating therapeutic controversies. The role of vaccines in the management of melanoma was discussed, including cell vaccines, dendritic cell-based vaccination and present research to improve the generation of melanoma vaccine-specific immunity. Adjuvant immunotherapy with high-dose IFN-alpha and an ongoing trial with biochemotherapy were debated. In addition, the role of chemotherapy and novel targeted agents in metastatic melanoma were discussed. Among the emerging agents and therapeutic targets presented were Bcl-2 antisense therapy, RAF kinases, heat-shock proteins, thalidomide and newer immunomodulatory drugs, cytotoxic T lymphocyte antigen-4 antibody and topical imiquimod. The symposium also provided an overview of existing and emerging agents and modalities in the management of patients with cutaneous T cell lymphomas, ocular melanoma and melanoma involving brain metastases. Sessions also included case-based learning and devoted ample time to providing 'how to' information for practising physicians. PMID- 15335330 TI - 4th European Stanley Conference on Bipolar Disorders, 23-25 September 2004, Aarhus, Denmark. Abstracts. PMID- 15335331 TI - Personality, identity styles, and religiosity: an integrative study among late adolescents in Flanders (Belgium). AB - The relationship between the two main dimensions of the religiosity domain (Exclusion vs. Inclusion of Transcendence and Literal vs. Symbolic) and both the Five-Factor Model of personality and Berzonsky's (1990) identity styles was investigated in a Flemish sample of late adolescents (N = 335). The results show that, whereas Exclusion vs. Inclusion is unrelated to any of the personality dimensions, Literal vs. Symbolic was strongly related to Openness to Experience and moderately to Agreeableness. Further, it was shown that Exclusion vs. Inclusion was positively related to the normative identity style, and that Literal vs. Symbolic correlated positively with the informational identity style and negatively with the diffuse/avoidant identity style. As expected, the relation between Openness to Experience and Literal vs. Symbolic was fully mediated by the informational identity style. Once Openness to Experience was taken into account, Agreeableness was no longer an important determinant of Literal vs. Symbolic. PMID- 15335332 TI - The stories we keep: autobiographical memory in American and Chinese middle-aged adults. AB - One hundred and eight European American and Chinese adults, aged between 38 and 60, participated in this questionnaire study. They each recalled 20 memories from any period of their lives. Memory content was analyzed as a function of culture (U.S. and China), life period (childhood, youth, early midlife, and peak midlife), and gender (female and male). Across the four life periods, Americans provided more memories of individual experiences and unique, one-time events and focused on their own roles and emotions. In contrast, Chinese were more inclined to recall memories of social and historical events and placed a great emphasis on social interactions and significant others in their memory narratives. Chinese also more frequently drew upon past events to convey moral messages than did Americans. In addition, memory content evidenced age-related increases in both autonomous and social orientations. Findings are discussed in light of the self definitional and directive functions of Autobiographical memory in the context of culture. PMID- 15335333 TI - Defensive copers show a deficit in passive avoidance learning on Newman's go/no go task: implications for self-deception and socialization. AB - The present study investigated whether passive avoidance learning was retarded by defensive coping strategies designed to minimize exposure to negatively valenced stimuli. High-anxious individuals, low-anxious individuals, and defensive copers completed a computerized go/no-go task, in which they learned when to press or not to press a button, in response to contingent positive and negative feedback. The duration that feedback remained onscreen was self-regulated. Defensive copers showed preferential reflection away from negative feedback, committed more passive-avoidance errors, and were characterized by impaired learning, overall. Further, the ratio of reflection on negative feedback to reflection on positive feedback directly mediated both passive-avoidance errors and overall learning. Defensive coping strategies, therefore, appear to interfere with passive avoidance learning, thereby fostering perseverative, dysfunctional action patterns by reducing knowledge gained from previous mistakes. Implications for the learning of effective socialization strategies, and for psychopathy-which is commonly characterized by similar passive-avoidance deficits-are subsequently considered. PMID- 15335334 TI - Gay and straight possible selves: goals, identity, subjective well-being, and personality development. AB - This study examined the relations of the salience and elaboration of gay and straight possible selves to subjective well-being (SWB) and ego development (ED) in a sample of 107 gay men and lesbians, who wrote narrative descriptions of their straight and gay best possible selves and rated the salience of these narratives (i.e., the clarity of the mental image, how easy it was to imagine). Independent raters coded the narratives for amount of elaboration (i.e., vivid detail). The salience of the gay possible self was positively related to SWB and outness, while the salience of the straight possible self was negatively related to SWB and being out. Furthermore, the possible self variables mediated the relationship of outness to well-being. Straight self-elaboration predicted personality development concurrently and prospectively, over 2 years. Results are interpreted as indicating that, while happiness may require us to focus upon our current life goals, personality development might require an exploration of the potential sacrifices of identity. PMID- 15335335 TI - The impact of personality on cognitive, behavioral, and affective political processes: the effects of need to evaluate. AB - Need to evaluate (NE) is a personality trait that reflects a person's proclivity to create and hold attitudes; people high in NE are especially likely to form attitudes toward all sorts of objects. Using data from the 1998 National Election Survey Pilot and the 2000 National Election Survey, NE was shown to predict a variety of important attitude-relevant cognitive, behavioral, and affective political processes beyond simply holding attitudes: NE predicted how many evaluative beliefs about candidates a person held, the likelihood that a person would use party identification and issue stances to determine candidate preferences, the extent to which a person engaged in political activism, the likelihood that a person voted or intended to vote, the extent to which a person used the news media for gathering information, and the intensity of emotional reactions a person felt toward political candidates. Thus, NE appears to play a powerful role in shaping important political behavior, emotion, and cognition. PMID- 15335336 TI - Match makers and deal breakers: analyses of assortative mating in newlywed couples. AB - We conducted a comprehensive analysis of assortative mating (i.e., the similarity between wives and husbands on a given characteristic) in a newlywed sample. These newlyweds showed (a) strong similarity in age, religiousness, and political orientation; (b) moderate similarity in education and verbal intelligence; (c) modest similarity in values; and (d) little similarity in matrix reasoning, self- and spouse-rated personality, emotional experience and expression, and attachment. Further analyses established that similarity was not simply due to background variables such as age and education and reflected initial assortment (i.e., similarity at the time of marriage) rather than convergence (i.e., increasing similarity with time). Finally, marital satisfaction primarily was a function of the rater's own traits and showed little relation to spousal similarity. PMID- 15335337 TI - Personality, threat, and cognitive and emotional reactions to stressful intercultural situations. AB - The present study examined individual differences in appraisal of and affective reactions to intercultural situations. A sample of 160 students filled out the Multicultural Personality Questionnaire (MPQ) and participated in an experiment in which they received a description of an intercultural situation that was either high or low in potential stressfulness. Individuals with high scores on the intercultural dimensions appraised the potentially stressful situation more positively and showed more positive and less negative reactions to the situation than did individuals with low scores on the MPQ. Interestingly, following a Terror Management Intervention (TMI), individual differences in emotional reactions to intercultural situations disappeared. The results could only partially be replicated using a general personality questionnaire, suggesting that these findings have at least some specificity to intercultural personality dimensions. PMID- 15335338 TI - Editorial: introduction to special section on face processing. PMID- 15335339 TI - Annotation: Development of facial expression recognition from childhood to adolescence: behavioural and neurological perspectives. AB - BACKGROUND: Intact emotion processing is critical for normal emotional development. Recent advances in neuroimaging have facilitated the examination of brain development, and have allowed for the exploration of the relationships between the development of emotion processing abilities, and that of associated neural systems. METHODS: A literature review was performed of published studies examining the development of emotion expression recognition in normal children and psychiatric populations, and of the development of neural systems important for emotion processing. RESULTS: Few studies have explored the development of emotion expression recognition throughout childhood and adolescence. Behavioural studies suggest continued development throughout childhood and adolescence (reflected by accuracy scores and speed of processing), which varies according to the category of emotion displayed. Factors such as sex, socio-economic status, and verbal ability may also affect this development. Functional neuroimaging studies in adults highlight the role of the amygdala in emotion processing. Results of the few neuroimaging studies in children have focused on the role of the amygdala in the recognition of fearful expressions. Although results are inconsistent, they provide evidence throughout childhood and adolescence for the continued development of and sex differences in amygdalar function in response to fearful expressions. Studies exploring emotion expression recognition in psychiatric populations of children and adolescents suggest deficits that are specific to the type of disorder and to the emotion displayed. CONCLUSIONS: Results from behavioural and neuroimaging studies indicate continued development of emotion expression recognition and neural regions important for this process throughout childhood and adolescence. Methodological inconsistencies and disparate findings make any conclusion difficult, however. Further studies are required examining the relationship between the development of emotion expression recognition and that of underlying neural systems, in particular subcortical and prefrontal cortical structures. These will inform understanding of the neural bases of normal and abnormal emotional development, and aid the development of earlier interventions for children and adolescents with psychiatric disorders. PMID- 15335340 TI - Face-memory and emotion: associations with major depression in children and adolescents. AB - BACKGROUND: Studies in adults with major depressive disorder (MDD) document abnormalities in both memory and face-emotion processing. The current study used a novel face-memory task to test the hypothesis that adolescent MDD is associated with a deficit in memory for face-emotions. The study also examines the relationship between parental MDD and memory performance in offspring. METHODS: Subjects were 152 offspring (ages 9-19) of adults with either MDD, anxiety disorders, both MDD and anxiety, or no disorder. Parents and offspring were assessed for mental disorders. Collection of face-memory data was blind to offspring and parent diagnosis. A computerized task was developed that required rating of facial photographs depicting 'happy,"fearful,' or 'angry' emotions followed by a memory recall test. Recall accuracy was examined as a function of face-emotion type. RESULTS: Age and gender independently predicted memory, with better recall in older and female subjects. Controlling for age and gender, offspring with a history of MDD (n = 19) demonstrated significant deficits in memory selectively for fearful faces, but not happy or angry faces. Parental MDD was not associated with face-memory accuracy. DISCUSSION: This study found an association between MDD in childhood or adolescence and perturbed encoding of fearful faces. MDD in young individuals may predispose to subtle anomalies in a neural circuit encompassing the amygdala, a brain region implicated in the processing of fearful facial expressions. These findings suggest that brain imaging studies using similar face-emotion paradigms should test whether deficits in processing of fearful faces relate to amygdala dysfunction in children and adolescents with MDD. PMID- 15335341 TI - Maternal personality and infants' neural and visual responsivity to facial expressions of emotion. AB - BACKGROUND: Recent investigations suggest that experience plays an important role in the development of face processing. The aim of this study was to investigate the potential role of experience in the development of the ability to process facial expressions of emotion. METHOD: We examined the potential role of experience indirectly by investigating the relationship between the emotional environment provided by mothers (as indexed by affective measures of their personality) and 7-month-olds' processing of emotional expressions (as indexed by visual attention and event-related potentials [ERPs]). RESULTS: For positive emotion, infants with highly positive mothers looked longer at fearful than happy expressions, and a subset of these infants who themselves also scored highly on positive temperament showed a larger negative central (Nc) component in the ERP to fearful than happy faces. For negative emotion, there were no detectable influences of maternal personality, although very fearful infants showed a larger Nc to fearful than happy expressions over the right hemisphere. CONCLUSION: To the extent that these variations in maternal disposition reflect variations in their expression of positive facial expressions, these results suggest that the emotional environment experienced by infants contributes to the development of their responses to facial expressions. PMID- 15335342 TI - Recognition of own-race and other-race faces by three-month-old infants. AB - BACKGROUND: People are better at recognizing faces of their own race than faces of another race. Such race specificity may be due to differential expertise in the two races. METHOD: In order to find out whether this other-race effect develops as early as face-recognition skills or whether it is a long-term effect of acquired expertise, we tested face recognition in 3-month-old Caucasian infants by conducting two experiments using Caucasian and Asiatic faces and a visual pair-comparison task. We hypothesized that if the other race effect develops together with face processing skills during the first months of life, the ability to recognize own-race faces will be greater than the ability to recognize other-race faces: 3-month-old Caucasian infants should be better at recognizing Caucasian faces than Asiatic faces. If, on the contrary, the other race effect is the long-term result of acquired expertise, no difference between recognizing own- and other-race faces will be observed at that age. RESULTS: In Experiment 1, Caucasian infants were habituated to a single face. Recognition was assessed by a novelty preference paradigm. The infants' recognition performance was better for Caucasian than for Asiatic faces. In Experiment 2, Caucasian infants were familiarized with three individual faces. Recognition was demonstrated with both Caucasian and Asiatic faces. CONCLUSIONS: These results suggest that (i) the representation of face information by 3-month-olds may be race-experience-dependent (Experiment 1), and (ii) short-term familiarization with exemplars of another race group is sufficient to reduce the other-race effect and to extend the power of face processing (Experiment 2). PMID- 15335343 TI - Face-sensitive cortical processing in early infancy. AB - BACKGROUND: Debates about the developmental origins of adult face processing could be directly addressed if a clear infant neural marker could be identified. Previous research with infants remains open to criticism regarding the control stimuli employed. METHODS: We recorded ERPs from adults and 3-month-old infants while they watched faces and matched visual noise stimuli. RESULTS: We observed similar amplitude enhancement for faces in the infant N290 and adult N170. In contrast, the infant P400 showed only a latency effect, making it unlikely to be the main precursor of the adult N170. CONCLUSIONS: We conclude that there is some degree of specificity of cortical processing of faces as early as 3 months of age. PMID- 15335344 TI - Event-related brain potentials reveal anomalies in temporal processing of faces in autism spectrum disorder. AB - BACKGROUND: Individuals with autism exhibit impairments in face recognition, and neuroimaging studies have shown that individuals with autism exhibit abnormal patterns of brain activity during face processing. The current study examined the temporal characteristics of face processing in autism and their relation to behavior. METHOD: High-density event-related brain potentials (ERPs) were recorded to images of faces, inverted faces, and objects from 9 individuals with autism spectrum disorder (15-42 years old) and 14 typical individuals (16-37 years old). RESULTS: With respect to a face-sensitive ERP component (N170), individuals with autism exhibited longer N170 latencies to faces than typical individuals but comparable latencies to objects. Typical individuals exhibited longer N170 latencies to inverted as compared to upright faces, whereas individuals with autism did not show differences in N170 latency to upright versus inverted faces. Neural speed of face processing, as reflected in N170 latency, correlated with performance on a face recognition task for individuals with autism. CONCLUSIONS: These data provide evidence for slowed neural speed of face processing in autism and highlight the role of speed of processing in face processing impairments in autism. PMID- 15335345 TI - Do children with autism perceive second-order relational features? The case of the Thatcher illusion. AB - BACKGROUND: This study presents two experiments that investigated whether children with autism were susceptible to the Thatcher illusion. Perception of the Thatcher illusion requires being able to compute second-order configural relations for facial stimuli. METHOD: In both experiments children with autism were matched for non-verbal and verbal ability with a group of children with moderate (non-specific) mental retardation (MLD) and a group of typically developing children respectively. Participants were asked to detect the 'unusual' face in a two-alternative-forced-choice version of the Margaret Thatcher illusion with grey-scale (Experiment 1) and monochrome 'Mooney' face images (Experiment 2). In Experiment 1 participants also performed a control task where buildings had been doctored in the same way as the facial stimuli. RESULTS: Children with autism were as susceptible to the Thatcher illusion as both control groups, in terms of accuracy and reaction time to make decisions about which face was unusual. Children with autism performed more accurately than children with MLD in the buildings task. CONCLUSION: Children with autism are able to compute second order configural features in faces and exhibit no difference in face processing, relative to appropriate control groups. PMID- 15335346 TI - Exploring the Williams syndrome face-processing debate: the importance of building developmental trajectories. AB - BACKGROUND: Face processing in Williams syndrome (WS) has been a topic of heated debate over the past decade. Initial claims about a normally developing ('intact') face-processing module were challenged by data suggesting that individuals with WS used a different balance of cognitive processes from controls, even when their behavioural scores fell within the normal range. Measurement of evoked brain potentials also point to atypical processes. However, two recent studies have claimed that people with WS process faces exactly like normal controls. METHOD: In this paper, we examine the details of this continuing debate on the basis of three new face-processing experiments. In particular, for two of our experiments we built task-specific full developmental trajectories from childhood to adolescence/adulthood and plotted the WS data on these trajectories. RESULTS: The first experiment used photos of real faces. While it revealed broadly equivalent accuracy across groups, the WS participants were worse at configural processing when faces were upright and less sensitive than controls to face inversion. In Experiment 2, measuring face processing in a storybook context, the face inversion effect emerged clearly in controls but only weakly in the WS developmental trajectory. Unlike the controls, the Benton Face Recognition Test and the Pattern Construction results were not correlated in WS, highlighting the different developmental patterns in the two groups. Again in contrast to the controls, Experiment 3 with schematic faces and non-face stimuli revealed a configural-processing deficit in WS both with respect to their chronological age (CA) and to their level of performance on the Benton. CONCLUSION: These findings point to both delay and deviance in WS face processing and illustrate how vital it is to build developmental trajectories for each specific task. PMID- 15335347 TI - The reliability and validity of tasks measuring perception of rapid sequences in children with dyslexia. AB - BACKGROUND: Claims that children with reading and oral language deficits have impaired perception of sequential sounds are usually based on psychophysical measures of auditory temporal processing (ATP) designed to characterise group performance. If we are to use these measures (e.g., the Tallal, 1980, Repetition Test) as the basis for intervention in language and literacy deficits, we need to demonstrate that they can effectively quantify individual differences. Therefore, questions of standardisation, reliability and construct validity can no longer be ignored. METHOD: We explored these issues in three studies: (i) 52 Dyslexics and Good Readers aged 8 to 11 years performed a task requiring perception of rapid sequences (PRS) based on the Tallal Repetition Test; (ii) a subgroup of the initial sample was retested on the task three to four months later, and after extended practice; (iii) a further subgroup then completed a rate of auditory processing task using a backward recognition masking paradigm. RESULTS: With a standardised methodology, we were able to replicate previous results with the PRS task, and demonstrate moderate reliability of measurement across time and practice. However, there were large effects of exposure and practice, and the task did not seem useful for identifying absolute and continuing deficits in given individuals. CONCLUSIONS: Our results call into question the use of this type of task as an individual measure of ATP. Neither is it certain that it is capturing what is currently understood as ATP. PMID- 15335348 TI - Parents and partners in crime: a six-year longitudinal study on changes in supportive relationships and delinquency in adolescence and young adulthood. AB - BACKGROUND: This study sought the answer to three questions: 1. Is having an intimate partner associated with the level of delinquency? 2. Does the quality of the relationship with an intimate partner, operationalised as partner support, predict the level of delinquency? 3. Does a relationship with an intimate partner or age moderate the association between parental support and delinquency? METHOD: Data from a three-wave, six-year longitudinal study of 1302 adolescents and young adults, aged 12-23 at wave 1, were used. RESULTS: 1. Univariate latent growth curve analysis showed that, as predicted, having an intimate partner does not lead to less criminality among young adults over the age of 20. We found no support for the hypothesis among the group of 12- to 20-year-old adolescents, since the group of mid-adolescents who consistently have a partner is more criminal than the other groups. 2. Our findings show that partner support is negatively related to criminality in both 12- to 20-year-olds and 21- to 23-year olds. The longitudinal effect of partner support is also uni-directional: partner support T1 certainly has an impact on criminality T3, but the reverse is not true. In both groups the influence of partner support is also greater than that of parental support. 3. Having an intimate partner moderates the association between parental support and delinquency, but in an age-specific manner. Parental support has no bearing on criminality when adolescents and young adults continuously have an intimate partner. Parental support does, however, cause a reduction in the level of criminality in adolescents and young adults who have never had a partner and adolescents who only have a partner at time 3. CONCLUSIONS: We interpret our results in terms of shifts in the relational system: if an adolescent finds a partner, that partner takes over the role of the parents in reducing criminality; if not, the parents remain important in doing so. PMID- 15335349 TI - Screening for DSM-IV externalizing disorders with the Child Behavior Checklist: a receiver-operating characteristic analysis. AB - BACKGROUND: This study examines the diagnostic accuracy of the CBCL syndrome AS scales for predicting DSM-IV Attention Deficit-Hyperactivity Disorder (ADHD) and Oppositional Defiant Disorder with or without Conduct Disorder (ODD/CD). METHODS: The sample included 370 children (187 probands and 183 siblings) participating in a family genetic study of attention and aggressive behavior problems. Univariate and stepwise logistic regression analyses were used to derive models for predicting two diagnostic conditions: ADHD and ODD/CD. RESULTS: The Attention Problems syndrome significantly predicted ADHD, and ODD/CD was significantly predicted by the Aggressive Behavior syndrome. Both scales demonstrated good diagnostic accuracy, as assessed through receiver operating characteristics analyses. Cut-point analyses confirmed the utility of low T-scores, 55 on the respective syndromes, for efficiently discriminating cases from noncases. CONCLUSIONS: CBCL syndromes display good diagnostic efficiency for assessing common externalizing disorders in children. PMID- 15335350 TI - Online aggressor/targets, aggressors, and targets: a comparison of associated youth characteristics. AB - BACKGROUND: While most youth report positive experiences and activities online, little is known about experiences of Internet victimization and associated correlates of youth, specifically in regards to Internet harassment. METHODS: The Youth Internet Safety Survey is a cross-sectional, nationally representative telephone survey of young regular Internet users in the United States. Interviews were conducted between the fall of 1999 and the spring of 2000 and examined characteristics of Internet harassment, unwanted exposure to sexual material, and sexual solicitation that had occurred on the Internet in the previous year. One thousand, five hundred and one regular Internet users between the ages of 10 and 17 years were interviewed, along with one parent or guardian. To assess the characteristics surrounding Internet harassment, four groups of youth were compared: 1) targets of aggression (having been threatened or embarrassed by someone; or feeling worried or threatened by someone's actions); 2) online aggressors (making rude or nasty comments; or harassing or embarrassing someone with whom the youth was mad at); 3) aggressor/targets (youth who report both being an aggressor as well as a target of Internet harassment); and 4) non harassment involved youth (being neither a target nor an aggressor online). RESULTS: Of the 19% of young regular Internet users involved in online aggression, 3% were aggressor/targets, 4% reported being targets only, and 12% reported being online aggressors only. Youth aggressor/targets reported characteristics similar to conventional bully/victim youth, including many commonalities with aggressor-only youth, and significant psychosocial challenge. CONCLUSIONS: Youth aggressor/targets are intense users of the Internet who view themselves as capable web users. Beyond this, however, these youth report significant psychosocial challenge, including depressive symptomatology, problem behavior, and targeting of traditional bullying. Implications for intervention are discussed. PMID- 15335351 TI - Current conceptualisation of body image dissatisfaction: have we got it wrong? AB - BACKGROUND: This study addresses limitations of previous research by examining the prevalence of body image dissatisfaction within two developmental periods: childhood and adolescence. METHODS: A sample of 448 boys and 508 girls completed self-report measures of global body image dissatisfaction. Weight and height of all participants were also determined. RESULTS: Our results indicated that body image dissatisfaction differs significantly depending upon sex and body mass. Importantly, the results revealed a multi-modal distribution in the data, particularly for boys. That is, for females there was a significant increase in body dissatisfaction across bodyweight, reflecting a predominant desire to be thinner. In contrast, for boys, there was a differential pattern; with those who were overweight wanting to be thinner, but those who were underweight wanting to be larger. Moreover, whilst for both sexes, body image dissatisfaction was found to be evident in childhood as well as adolescence, during the former developmental period, it appeared to be less pronounced. CONCLUSIONS: These findings indicate that, contrary to previous propositions, body image dissatisfaction is of concern for males as well as females, although the distribution of such is strikingly different. The outcomes suggest that it is time to reframe our conceptualisation of this construct. Specifically, the identified non-linear distribution of means indicates that inferential analysis of body image dissatisfaction data needs to be conducted independently for each sex, as well as each body mass grouping. PMID- 15335352 TI - Characterization of Ets-binding sequence of human transglutaminase 3 gene promoter. AB - The Ets- and Sp1-transcription factors have been implicated in the epithelial specific expression of transglutaminase 3 gene (TGM3) in vitro. By electrophoretic mobility shift assay (EMSA), the core motif of Ets-binding sequence of TGM3 was determined as ACAGGAAT (-118 to -111 bp from transcriptional start site). However, a sequence extending from -120 to +10 of TGM3 proximal promoter region failed to induce the expression of CAT reporter in transfected normal human epidermal keratinocytes (NHEKs). In contrast, a construct extending from -126 to +10 bp showed the highest expression of CAT gene, indicating the presence of an important element(s) between -126 and -120 bp that affects TGM3 expression in NHEKs. To find the critical sequences in this promoter region, we performed EMSA analysis with competitor oligonucleotides, in which the upstream nucleotides of Ets core motif were mutated. The results showed that the mutation of A at -125 dramatically reduced the binding of Ets to its recognition sequence. Transient transfection assay revealed that the mutation in this nucleotide greatly diminished the expression of CAT reporter gene in NHEKs. Together, these results suggest that the upstream sequence of Ets core motif is critical for the expression of TGM3 in NHEKs cultured in vitro. PMID- 15335353 TI - Differential expression of mast cell characteristics in human myeloid cell lines. AB - In order to better understand the mechanisms governing display of mast cell characteristics in human myeloid cells, we have studied the mast cell phenotype in human promyelocytic (HL-60) and myelocytic (U-937, TPH-1) vs. basophilic (KU 812) and mast cell (HMC-1) lines, in part also in skin mast cells and blood monocytes, at mRNA and protein level before and after stimulation with mast cell growth factors. In unstimulated cells, mRNA for the stem cell factor (SCF) receptor c-kit and the gamma chain of the high-affinity IgE receptor (FcepsilonRI) was noted in all cells studied. Like mast and basophilic cells, THP 1 cells expressed the FcepsilonRIalpha and beta chains and weakly histidine decarboxylase (HDC), but they lacked mRNA for mast cell-specific proteases [tryptase, chymase, carboxypeptidase A (CPA)]. In contrast, HL-60 and U-937 cells lacked FcepsilonRIalpha, but expressed tryptase and chymase, HL-60 cells also CPA. KU-812 cells failed to express the basophil-specific marker 2D7. After a 10 day culture with SCF or fibroblast supernatants, baseline mRNA expression of most mast cell characteristics was upregulated, whereas c-kit mRNA expression decreased in all but THP-1 cells. Differential mRNA expression of FcepsilonRI vs. protease (tryptase) was confirmed at protein level by immunocytochemistry and enzymatic activity. KU-812 cells are thus closest to skin mast cells in that they express all molecules studied, except for chymase, followed by THP-1 cells that lack all mast cell proteases. In contrast, HL-60 and U-937 cells fail to express the FcepsilonRIalpha and beta chains but express most mast cell proteases. The selective and differential expression of mast cell characteristics in human myeloid cell lines suggests that induction of the mast cell phenotype is regulated by several independent genes and that mast cells and basophils branch off at early and distinct points of myeloid development. PMID- 15335354 TI - Expression of psoriasis-associated fatty acid-binding protein in senescent human dermal microvascular endothelial cells. AB - Aging is associated with the progressive pathophysiologic modification of endothelial cells. In vitro endothelial cell senescence is accompanied by proliferative activity failure and by perturbations in gene and protein expressions. Moreover, this cellular senescence in culture has been proposed to reflect processes that occur in aging organisms. In order to observe the changing patterns of protein expression in senescent human dermal microvascular endothelial cells (HDMECs), proteins obtained from both early- and late-passaged HDMECs were separated by two-dimensional electrophoresis, visualized by silver staining, and quantified by image processing. Proteins of interest were extracted by in-gel digestion with trypsin and quantified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), by searching the National Center for Biotechnology Information protein-sequence database. More than 2000 spots were detected by 2D electrophoresis within a linear pH range of 3-10. Twenty-two major differentially expressed spots were observed in serially passaged HDMECs and identified with high confidence by MALDI TOF-MS. One of these spots was found to be a 14-15 kDa psoriasis-associated fatty acid-binding protein (PA-FABP) with high affinity for long-chain fatty acids. The expression of PA-FABP was confirmed to be elevated in senescent HDMECs (passage 20) by fluorescence-activated cell sorting (FACS), confocal laser microscopy, and by immunohistochemistry in aged human skin tissue. Our results suggest that the overexpression of FABP in cultured senescent HDMECs is closely related to skin aging. PMID- 15335355 TI - TARC augments TNF-alpha-induced CTACK production in keratinocytes. AB - Thymus- and activation-regulated chemokine (TARC)/CCL17 and cutaneous T cell attracting chemokine (CTACK)/CCL27 are both pivotal mediators of the inflammatory reaction of atopic dermatitis (AD). TARC attracts CCR4 positive T cells known to be mainly of Th2 subtype whereas CTACK attracts skin-homing T cells of both Th1 and Th2 subtype that express CCR10. We found that CTACK can be induced in cultured human keratinocytes by tumor necrosis factor-alpha (TNF-alpha), but not by TARC alone. However, if the keratinocytes were preincubated with TNF-a for 6 h, TARC was able to augment the CTACK-inducing effect of TNF-a. Performing immunohistochemical stainings, reverse-transcription polymerase chain reaction (RT-PCR), and Western blotting, we found that TNF-a-induced CCR4 mRNA production, but that stimulated as well as non-stimulated keratinocytes expressed CCR4. In order to see if these results had any clinical relevance, we investigated the plasma concentrations of TARC and CTACK from 48 patients suffering from AD. This revealed that TARC and CTACK concentrations in plasma correlate with each other. Surprisingly, p-CTACK correlated inversely with SCORAD scores of the patients, which most likely is due to the treatment the patients received. Our results suggest that the primary Th2-dominated inflammatory reaction in AD induced by TARC leads to an augmented skin-specific inflammatory reaction through CTACK. PMID- 15335356 TI - Topical beta-carotene is converted to retinyl esters in human skin ex vivo and mouse skin in vivo. AB - Human epidermis contains endogenous retinoids (retinol and retinyl esters) and carotenoids (mostly beta-carotene). Previous studies have shown that the enzymes involved in retinoid metabolism are present in human epidermis. There is still a controversy about the presence in the skin of the enzymes able to convert beta carotene into vitamin A (retinol), although a recent study demonstrated the conversion of beta-carotene into retinol in human cultured epidermal cells. In this study, we addressed the question of the possible bioconversion of topical beta-carotene into vitamin A or derivatives by human and mouse skin. Surgically excised human abdominal skin was mounted on Franz perfusion chambers to assess the cutaneous penetration of topical beta-carotene as well as its metabolism, after a 24-h incubation period, whereas hairless mice received topical beta carotene 24 h before assaying epidermal beta-carotene and retinoid concentrations. Epidermal retinoid and beta-carotene concentrations were determined by high-pressure liquid chromatography. Topical beta-carotene penetrated well into human and mouse epidermis and induced a 10-fold (human) and a threefold (mouse) increase of epidermal retinyl esters, which demonstrates that topical beta-carotene is converted into retinyl esters by human and mouse epidermis and thus appears as a precursor of epidermal vitamin A. PMID- 15335357 TI - Selective cultivation of normal human sebocytes in vitro; a simple modified technique for a better cell yield. AB - Selective cultivation of normal human sebocytes is essential for better understanding of drug pharmacokinetics and diseases of the pilosebaceous apparatus. In the present study, sebocytes are selectively cultivated in vitro using modified MCDB 153 medium to which cholera toxin (1 x 10(-9) M), crude bovine pituitary extract (70 micro g/ml), epidermal growth factor (10 ng/ml), basic fibroblast growth factor (2 ng/ml), hydrocortisone (1.4 x 10(-6) M), insulin (10 micro g/ml), fetal bovine serum (10%), and antibiotics were added. To maintain contact of the floating sebaceous lobules with culture plate, two methods have been adopted. (i) Using little amount of culture medium that barely covers the gland lobules with frequent dropping of the medium to replace the loss by evaporation (almost every 2 h). (ii) Placing a sterile glass slide cover over the gland lobules in the presence of enough culture medium. Both methods are performed for the first 72 h of inoculation, when cells are seen outgrowing from sebaceous lobules. Both populations show the characteristic morphology of sebocytes in culture, namely polygonal shape with abundant cytoplasm resembling basal keratinocytes. As the culture grows older, a vacuolated refractile cytoplasm becomes evident. A comparison of both methods revealed a significant percentage increase of sebocytes obtained from covered lobules being 144% (P = 0.02) on day 4, 162% (P = 0.009) on day 8, and 173% (P < 0.001) on day 12 of incubation. No further proliferation is measured thereafter. Cells obtained from both methods also showed no difference in lipogenesis (Oil-Red stain) or in the expression of the specific epidermal membrane antigen as shown by monoclonal antibody labeling (alkaline phosphatase anti-alkaline phosphatase-technique). To conclude, covering the sebaceous glands during the first 72 h of primary culture provides an excellent contact with the culture plate and hence a significant better yield of sebocytes more suitable for large experimental work. PMID- 15335359 TI - Abstracts of the 8th Congress of the European Federation of Neurological Societies. Paris, France, 4-7 September 2004. PMID- 15335361 TI - Fibrin glue in the management of anal fistulae. AB - OBJECTIVE: Fibrin glue has been used as a sphincter sparing approach for the treatment of anal fistulae for two decades. However, there is uncertainty about its short and long-term efficacy. The objective of this review was to ascertain the role of fibrin glue in the management of anal fistulae, including assessment of recurrence rates, continence disturbance and other complications. METHODS: We searched Medline (January 1966 to February 2004), the Cochrane database, and EMBASE using the terms anal fistulae, fistula-in-ano, and fibrin glue. Relevant papers from the reference lists of these articles and from the authors' personal collections were also reviewed. A systematic review of all articles relating to the use of fibrin glue in the treatment of anal fistulae was performed. This included 19 studies. Reviewers performed data extraction independently. Outcomes evaluated included recurrence rates, continence disturbance, septic complications, adverse drug reactions, and duration of follow-up. Heterogeneity of the clinical trials made direct comparisons difficult and meta-analysis impossible. RESULTS: The success rates reported in published studies range from 0% to 100%. Differences in patient selection (including fistula aetiology and type), treatment protocols, and follow-up duration may contribute to such diverse results. CONCLUSIONS: Fibrin glue is simple to use, has a minimal morbidity and should not affect later treatment options in the event of its failure. It is therefore theoretically attractive as a first line treatment in the management of those types of anal fistula in which it has been shown to work. However, further research into 'biological' glues is merited and these subject to randomised controlled study. PMID- 15335362 TI - In vitro biopsy of colorectal carcinomas. AB - OBJECTIVE: To determine the best part of an ulcerated colorectal neoplasm from which to take a biopsy in order to prove that the lesion is invasive. PATIENTS AND METHODS: In this experimental study, biopsies were obtained from 50 ulcerated colorectal tumours using colonoscopic biopsy forceps on the resected specimen after major colorectal resections. Four biopsy sites were chosen, the junction between the normal mucosa and the rolled edge, the top of the rolled edge, the junction between the rolled edge and the ulcer base, and the centre of the ulcer. A code was used so that the pathologist was not aware of the site of each biopsy. RESULTS: Biopsies from the very edge of the lesion were positive for carcinoma in 16% of cases, from the top of the roll in 64%, from the inner aspect of the roll in 88% and from the centre of the ulcer in 90%. CONCLUSION: Biopsies from ulcerated colorectal carcinomas are more likely to be positive if taken from the centre of the lesion, rather than the very edge. PMID- 15335363 TI - The quality of patient-orientated internet information on colorectal cancer. AB - OBJECTIVE: The Internet provides an enormous amount of patient-orientated information on colorectal cancer. This study examined its accessibility and quality. METHODS: Sites were identified using two search engines (Google and Hotbot) and the search terms 'bowel cancer' and 'colon and rectal cancer'. The first 100 sites in each search were visited and classified. Sites that contained patient information were assessed using the 'Discern' instrument for reliability and quality of information on treatment choices. RESULTS: Google identified 55,700 sites for the search term 'colorectal cancer' and 214,000 for 'bowel cancer'. Hotbot produced 27,700 and 190,000, respectively. Four hundred sites were studied. Only 25 (6%) sites were duplicated in the searches. One hundred and eighteen (30%) provided information, 70 (18%) were lists of links, 27 (7%) were adverts, 22 (6%) promoted medical centres, 51 (13%) were dead links, 15 (4%) were message boards. Of the 118 that provided information 73 (62%) advised on treatment and 73 (62%) were designed for patients. The sources of information were clear in 55 (47%) and the date when this information was reported was given in only 63 (53%). By adapting the Discern instrument, sites were classified as excellent 18 (15.3%), very good 19 (16.1%), good 28 (23.7%), fair 8 (6.8%) and poor 45 (38.1%). CONCLUSIONS: The Internet provides a wealth of information on colorectal cancer but the best sites are difficult for patients to distinguish from the thousands of sites returned by search engines. Clinicians should guide patients to quality sites to avoid confusion and misinformation. PMID- 15335364 TI - Reliability of radiographic measurement of small intestinal length. AB - OBJECTIVE: The capacity for absorption of nutrients after small intestinal resection is related to the remaining length of intestine, and short bowel syndrome has major implications. Accurate pre-operative determination of bowel length may be useful in conditions where further resection of small bowel is contemplated. The aim of this study is to evaluate the accuracy of small bowel length measured by barium follow-through (BaFT) examination. PATIENTS AND METHODS: Twenty-two patients underwent both intra-operative measurement of small bowel length, and BaFT examination where length was measured using an opisometer. RESULTS: The difference between operative and radiological measurement (mean +/- standard deviation: 15.7 +/- 16 9%) was least in those with the shorter small intestine (overall variation: r = 0.703, P < 0.01, regression coefficient: 0.118, intercept: 15.1). There was no difference in whether BaFT was performed pre or postoperatively. CONCLUSION: Assessment of small bowel length by BaFT radiography is accurate, particularly in those with shorter small bowel lengths. PMID- 15335365 TI - Nicorandil associated anal ulceration. AB - OBJECTIVE: Nicorandil is a vasodilator used to control angina. It has been associated with oral and anal ulceration that resolves upon withdrawal of the drug. PATIENTS AND METHODS: We report a series of 13 patients with nonspecific anal ulceration all of whom were receiving nicorandil for control of symptoms of ischaemic heart disease. RESULTS: All the ulcers had similar clinical and histological appearances. All the ulcers healed on withdrawal of the drug. CONCLUSION: Nicorandil-associated anal ulceration should be considered in the differential diagnoses of nonhealing anal ulcers. PMID- 15335366 TI - Benefits of 'clean sweep' in Peutz-Jeghers patients. AB - OBJECTIVE: Laparotomy is the treatment of choice in Peutz-Jeghers Syndrome (PJS) patients for endoscopically irretrievable symptomatic polyps and polyp-related complications. During the last decade, we have operated on majority of the PJS patients with the purpose of removing all the gastrointestinal polyps (clean sweep), when an operation was indicated. The aim of this study is to evaluate the effect of clean sweep technique on the need for repeated surgery compared to a problem focused approach. PATIENTS AND METHODS: All patients with PJS treated in our institution since 1964 were studied. They were placed into two groups; those who had a problem-focused operation and those who were operated with the purpose of removing all small and large intestinal polyps. Demographics, presentation, follow-up period and the need for recurrent surgery were compared. RESULTS: We identified 11 patients (4 males, 7 females). Eight patients (5 females; median age 18.5) had problem-focused surgery for bleeding-anaemia (n = 3) or obstruction intussusception (n = 5). These patients required 23 further operations within 87 patient-follow-up-years (2.64 operations per 10 years). Three patients (2 females; median age 6) were operated for bleeding-anaemia (n = 1) or obstruction intussusception (n = 2) using the 'clean sweep' approach. These patients did not require any further surgery within 21 patient-follow-up-years. The gender, presentation and follow-up periods were similar between the groups. However, the 'clean sweep' technique appears to have reduced the need for further operations when it is compared with problem-focused approach (P = 0.01). CONCLUSION: To reduce the need for abdominal surgery and consequent problems in PJS patients, an attempt to remove all detected polyps (clean sweep technique) may be beneficial in these patients. PMID- 15335367 TI - Transanal endoscopic microsurgery: clinical and functional results. AB - OBJECTIVE: Transanal endoscopic microsurgery (TEM) has become increasingly common in the management of rectal adenomas and also in selected cases of rectal carcinomas. The aim of this study was to assess the results in a consecutive series of patients after introducing the TEM technique. PATIENTS AND METHODS: All 58 patients operated with TEM from January 1996 to January 1999 were evaluated in a retrospective review. Forty-eight patients answered a clinically validated questionnaire a median of 22 months after TEM. Eighty patients who had undergone transanal excision and 12 who had undergone York Mason's procedure served as a reference group with respect to recurrence rates. RESULTS: The complication rate was 5% (immediate) and 14% (long-term). The overall 30-day mortality rate was zero. An impairment of continence was seen in 18 (37%) patients. Of these, all 18 experienced varying degree of incontinence to liquid stool, 14 also to flatus and 5 of them even to solid stool. The recurrence rate was 11% in adenomas and 14% in cancers; T1, 1 (10%) recurrence and T2, 1 (50%) recurrence. There was a correlation between operating time and impairment of continence as well as recurrence rate. CONCLUSION: TEM is a safe procedure, having a low recurrence rate and an acceptable functional outcome. PMID- 15335368 TI - Anorectal physiology in relation to clinical subgroups of patients with severe constipation. AB - OBJECTIVE: The aim of this study was to evaluate anorectal physiology in relation to clinically defined subgroups of patients with idiopathic constipation and to analyse relationships between anorectal physiology and rectal evacuation. SUBJECTS AND METHODS: One hundred consecutive patients with idiopathic constipation were clinically categorized as slow transit (n=19), outlet obstruction (n=52) and a group with mixed symptoms (n=29). They were examined by recording anal pressures and also rectal volumes in response to stepwise increases in rectal pressure (5-60 cm H2O). The manovolumetric results were compared with 28 sex and aged matched controls. Rectal evacuation was measured by computer-based image analysis of rectal emptying rate in defaecography. RESULTS: The rectal pressure thresholds for filling, urge and pain did not differ between the groups but there were proportionally more patients in the slow transit and mixed group with thresholds for filling exceeding 25 cm H2O (P=0.04). In total, 18% of patients had impaired sensitivity which was associated with long duration of symptoms (P < 0.05). Patients with grossly impaired rectal sensitivity (filling threshold > 40 cm H2O) had impaired rectal evacuation (P < 0.05). The rectal compliance was increased in the slow transit and mixed group (P < 0.01 0.05) in the pressure interval 5-15 cm H2O. Anal resting and squeeze pressures did not differ between the groups although 7/19 in the slow transit group had values around the lower limit of controls. Slow wave frequency was lower in all patient groups (P < 0.001 vs. controls). Rectal evacuation was not related to sphincter function or to rectal compliance. CONCLUSIONS: Clinical categorization of constipated patients defines groups where altered anorectal physiology is not uncommon. Constipation with symptoms of infrequent defaecation may be associated with impaired rectal sensitivity and increased rectal compliance whereas outlet obstruction symptoms are not clearly related to changes in anorectal physiology. PMID- 15335369 TI - Transanal endoscopic microsurgery: indications and results after 100 cases. AB - OBJECTIVE: Transanal endoscopic microsurgery (TEM), a minimally invasive technique has been employed in the excision of benign and well-selected malignant rectal tumours since June 1998. We present a prospective descriptive study and analyse the currently accepted indications. PATIENTS AND METHODS: Over a 4-year period 100 patients underwent TEM for treatment of rectal tumours located between 4 and 18 cm from the anal verge. RESULTS: TEM was performed in 71 cases for adenomas, 20 potentially curative excisions for pre-operative staged low-grade carcinoma, 3 palliative procedures for advance carcinoma, 5 carcinoids and 1 solitary ulcer. The local complication rate included wound breakdown in 7 patients, three of them requiring ileostomy. Conversion to laparotomy was performed in two patients. Five adenomas recurred and were successfully treated by TEM. Of the cancers, four patients required immediate salvage therapy by means of total mesorectal excision. Three patients underwent palliative TEM procedures combined with radiotherapy. A single cancer recurrence was treated by means of abdomino-perineal resection after radiotherapy. CONCLUSIONS: TEM appears to be an effective method of excising benign tumours and selected T1 carcinomas of the rectum. The superior exposure of tumours higher in the rectum combined with the greater precision of excision make this minimally invasive technique an attractive surgical approach. PMID- 15335370 TI - What factors affect lymph node yield in surgery for rectal cancer? AB - OBJECTIVE: The detection of lymph node metastases is of vital importance in patients undergoing excisional surgery for rectal cancer as it provides important prognostic information and facilitates decision-making with regards to adjuvant therapy. It has been suggested that patients in whom only a small number of nodes are present in the excised specimen have a worse prognosis, presumably due to inadequate lymphadenectomy and consequent understaging of the disease. The aim of this study was to determine which factors affect the yield of lymph nodes. METHODS: This was a retrospective study of patients who had undergone a resection for histologically proven adenocarcinoma of the rectum. The total number of lymph nodes identified in the excised specimen was recorded in each case. A multivariate analysis was performed to ascertain whether this number was significantly influenced by any of several variables. RESULTS: A total of 167 patients were studied (M:F ratio 107 : 60, median age 70 years). The median number of lymph nodes contained within the resected specimen was 16 (interquartile range 10-21). On univariate analysis a significantly higher yield of lymph nodes was obtained with tumours in the middle third of the rectum (P=0.007), larger tumours (P < 0.001), more locally advanced tumours according to both pT staging (P=0.001) and Dukes' staging (P=0.020), an increased number of involved nodes (P=0.003) and examination by a specialist histopathologist (P=0.003). On multivariate analysis the only significant variables were tumour size (P=0.021), number of positive nodes (P=0.007) and histopathologist (P=0.021). CONCLUSIONS: The number of lymph nodes identified within the excised specimen in patients undergoing resection of a rectal cancer positively correlates with the size of the tumour and is also dependent on the examining histopathologist. In addition, in node-positive patients the number of involved nodes increases with increasing lymph node yield. PMID- 15335371 TI - Long-term results of the antegrade continent enema procedure for constipation in adults. AB - OBJECTIVE: The aim of this study was to evaluate the long-term results of the Antegrade Continent Enema (ACE) procedure for treating severe constipation in adults. METHODS: Over 10 years 37 ACE conduits were created in 32 patients (median age 35 years, 26 women) with constipation caused by slow transit, obstructed defaecation or both. Conduits were created from the appendix (n = 20, 54%), ileum (n = 10, 27%), neoappendix caecostomy (n = 5, 14%) or colon (n = 2, 5%). Clinical records were retrospectively reviewed to determine outcome. RESULTS: After a median follow up of 36 (range 13-140) months, 28 (88%) required at least one further procedure on a primary conduit, including reversal in 19 (59%). Five patients had a second conduit fashioned, two successfully. Conduit type and constipation cause did not significantly influence the rates of ACE reversal or major revision. Ileal conduits were associated with fewer minor revision procedures for stenosis (1 in 7 patients) than appendix conduits (21 in 20 patients). There was one (3%) serious complication. Satisfactory ACE function was ultimately achieved in 47% of patients, at last follow up. After ACE reversal, 9 (28%) patients underwent formation of an end stoma and 3 patients had a colectomy. CONCLUSIONS: Revision procedures are common, but approximately half of patients can expect satisfactory long-term ACE function. ACE conduit reversal does not preclude subsequent alternative surgical strategies to treat this difficult condition. PMID- 15335372 TI - Endoscopic morphological anticipation of submucosal invasion in flat and depressed colorectal lesions: clinical implications and subtype analysis of the kudo type V pit pattern using high-magnification-chromoscopic colonoscopy. AB - OBJECTIVE: Focal submucosal invasive colorectal cancers (submucosa-sm1) can be managed by endoscopic mucosal resection (EMR) as local lymph node metastasis (LNM) are rare. Lesions are usually flat, depressed or mixed. In deeper vertical submucosal invasion (sm2-3) LNM rates exceed 10-15%. EMR within this group can be complicated by perforation, noncurative resection and may leave LNM untreated. It is therefore essential to differentiate accurately focal sm1 disease from submucosal sm2/3 disease. The aim of this study was to evaluate the relationship between the invasive type V pit pattern using high-magnification-chromoscopic colonoscopy (HMCC) and submucosal invasive depth for flat and depressed colorectal lesions. METHODS: Total colonoscopy was performed by a highly selected single endoscopist using the Olympus C240Z on 850 patients between January 2001 and July 2003. Kudo type V pits were identified using 0.05% crystal violet (CV) applied directly to the lesion using a steel tipped catheter. Type V pits were graded into class V(n)A-C as described by Nagata. Morphology was documented using the Japanese Research Society classification (JRSC). Histological sections, with reference to mucosal invasive characteristics, acquired using EMR or surgical excision were then compared with the pit pattern. RESULTS: Fifty-one lesions showed a type V pit pattern. The kappa coefficient of agreement between pit the type V pit pattern and histologically confirmed submucosal invasion was 0.51 (95% CI). Following resection, 97% of lesions were correctly anticipated to have sm2 + invasion using pit type Vn(B) and Vn(C) as clinical indicators of invasive disease. Specificity was low at 50% with an accuracy of 78%. CONCLUSIONS: The type V pit pattern is useful for the in vivo staging of submucosal invasive depth in flat and depressed colorectal lesions and is as sensitive as conventional 7.5 MHz EUS. There was a tendency to over-stage lesions and hence the technique is limited by its low overall specificity. PMID- 15335373 TI - 'An investigation of the nervous control of defecation' by Denny-Brown and Robertson: a classic paper revisited. 1935. AB - In 1935 two young neurologists, Derek Denny-Brown and E. Graeme Robertson, published an article explaining the mechanisms underlying human defaecation based on a manometric study in patients with sacral root and spinal cord lesions, and normal subjects. This article is still routinely cited in studies of rectal and sphincter ani function. Unfortunately, however, the article itself is not written well, being composed of long convoluted sentences and containing 79 often indecipherable figures. Difficult-to-understand articles were common to the publications of Denny-Brown, who became one of the most prominent neurologists of the twentieth century. In accord with our prior work explaining Denny-Brown and Robertson's earlier paper on micturition, we provide here what we hope is a clear explanation of the methods and results in their study on defaecation. PMID- 15335376 TI - Coloproctology in the Czech Republic. PMID- 15335377 TI - Coloproctology in Central Europe. PMID- 15335378 TI - Coloproctology in Hungary. PMID- 15335379 TI - Coloproctology in Lithuania. PMID- 15335380 TI - Coloproctology in Poland. PMID- 15335381 TI - Coloproctology in Serbia and Montenegro. PMID- 15335385 TI - Techniques for the removal of self-expanding metallic colorectal stents. PMID- 15335386 TI - Colonic pouchography is not routinely required prior to stoma closure. PMID- 15335387 TI - Poor clinical results after surgery for occult rectal prolapse. PMID- 15335390 TI - Review article: definition and diagnosis in portal hypertension--continued problems with the Baveno consensus? AB - The goals of the Baveno workshops were to develop consensus definitions of key events related to portal hypertension and variceal bleeding, and to produce guidelines to facilitate the conduct and reporting of clinical trials. The consensus definitions concern the diagnosis of active bleeding, failure to control bleeding, the criteria to distinguish continuing bleeding from rebleeding, and the means of assessing failure to prevent rebleeding. The guidelines concern the timing of diagnostic endoscopy, the policy for blood volume restitution, the measures to prevent infection and encephalopathy, and the treatment options for acute bleeding, as well as primary and secondary prophylaxis. The intention of the experts who developed the guidelines was that, as feedback from their practical application develops, they should be adapted to better fit the practical needs. The applicability of the Baveno definitions has been evaluated in a study where the definitions of clinically significant bleeding, failure to control bleeding, the time frame for the acute bleeding episode and the definition of rebleeding were tested. The main criticism raised in this study was that tachycardia, one of the criteria that define failure to control bleeding, was misleading in 15% of patients who had the symptom but were not bleeding. PMID- 15335392 TI - Review article: the relevance of portal pressure and other risk factors in acute gastro-oesophageal variceal bleeding. AB - Gastro-oesophageal variceal bleeding is the last step in a chain of events that starts with an increased portal pressure, and is followed by the formation and progressive dilatation of gastro-oesophageal varices. When the tension of the thin wall of the varices exceeds its elastic limit, the varices rupture and bleed. Wall tension is directly proportional to variceal pressure (which is a function of portal pressure) and variceal radius, and inversely related to the thickness of the variceal wall. The above facts explain why a high portal pressure (usually determined by the hepatic venous pressure gradient, or HVPG) and the presence at endoscopy of large varices with red wheals, red spots or diffuse redness on the varices (signalling a reduced wall thickness) correlate with the risk of bleeding. PMID- 15335394 TI - Review article: a critical comparison of drug therapies in currently used therapeutic strategies for variceal haemorrhage. AB - Vasoactive drugs are safe and easy to administer, and universal treatment is the first-line approach for all patients with suspected variceal bleeding. There are strong arguments that the combination of vasoactive drugs, started as soon as possible, and endotherapy later on is the best therapeutic option, particularly in cases of ongoing bleeding at the time of endoscopy. The main action of vasoactive drugs is to reduce variceal pressure. This can be achieved by diminishing the variceal blood flow and/or by increasing resistance to variceal blood flow inside the varices. Changes in variceal pressure parallel changes in portal pressure. Drugs for the treatment of variceal bleeding can therefore be assessed by measuring the changes in portal pressure, azygos blood flow and variceal pressure. Vasoactive drugs can be divided into two categories: terlipressin (Glypressin), and somatostatin and its analogues, especially octreotide. Terlipressin significantly reduces portal and variceal pressure and azygos flow, is superior to placebo in the control of variceal haemorrhage and improves mortality. It is beneficial when combined with sclerotherapy. It also has the advantage that it might preserve renal function, one of the most important factors affecting the outcome of cirrhosis. As such, terlipressin is the most potent of the various vasoactive drugs. Somatostatin significantly reduces portal and variceal pressure and azygos flow, is superior to placebo in controlling variceal haemorrhage, and improves the success of sclerotherapy. The effect of octreotide is well established for preventing the increase in portal pressure after a meal (similar to blood in the intestines) though the effect of ocreotide on variceal pressure is controversial. PMID- 15335396 TI - Review article: hepatorenal syndrome--definitions and diagnosis. AB - Hepatorenal syndrome (HRS) is a severe complication of cirrhosis that develops in the final phase of the disease. Two types of HRS exist. Type 1 is defined by a rapid reduction of renal function and in type 2 HRS the reduction of renal function is slowly progressive. Type 1 HRS is diagnosed when the serum creatinine level increases by more than 50% of the baseline value to above 133 micromol/L. According to the International Ascites Club, HRS is defined by the presence of five criteria: (1) severe cirrhosis; (2) glomerular hypofiltration; (3) no other functional or organic causes; (4) failure of plasma volume expansion; (5) no proteinuria. Additional diagnostic criteria may be present. The diagnosis of HRS may be difficult in patients with severe cirrhosis. Other types of acute renal failure may occur. For example, ischaemic or toxic tubular necrosis or sepsis may cause renal failure in these patients. Furthermore, uncontrolled HRS may lead to ischaemic tubular necrosis; thus, these patients must be managed as soon as possible in an intensive care unit. PMID- 15335398 TI - Review article: pathogenesis and pathophysiology of hepatorenal syndrome--is there scope for prevention? AB - The hepatorenal syndrome (HRS) is a functional impairment of the kidneys in chronic liver disease caused by a circulatory failure. The prognosis is poor, particularly with type 1 HRS, but also type 2, and only liver transplantation is of lasting benefit. However, recent research into the pathophysiology of ascites and HRS has stimulated new enthusiasm in their prevention and treatment. Patients with HRS have hyperdynamic circulatory dysfunction with reduced arterial blood pressure and reduced central blood volume, owing to preferential splanchnic arterial vasodilatation. Activation of potent vasoconstricting systems, including the sympathetic nervous and renin-angiotensin-aldosterone systems, counteracts the arterial vasodilatation and leads to a pronounced renal vasoconstriction with renal hypoperfusion, a reduced glomerular filtration rate, and intense sodium water retention. Thus prevention of HRS should seek to improve liver function, limit arterial hypotension and central hypovolaemia, and reduce renal vasoconstriction and the renal and interstitial pressures. Portal pressure can be reduced with beta-adrenergic blockers and transjugular intrahepatic portosystemic shunt (TIPS). Precipitating events, like infections, bleeding, and postparacentesis circulatory syndrome, should be treated to avoid further circulatory failure. Improvement in arterial blood pressure and central hypovolaemia can be achieved with vasoconstrictors, such as terlipressin (Glypressin), and plasma expanders such as human albumin. In the future endothelins, adenosine antagonists, long-acting vasoconstrictors, and antileukotriene drugs may play a role in preventing and treating HRS. PMID- 15335400 TI - Review article: prognosis of hepatorenal syndrome--has it changed with current practice? AB - The Consensus Conference on Hepatorenal Syndrome (HRS) organized by the International Ascites Club in 1994 redefined HRS, introduced new diagnostic criteria that are now widely accepted, and proposed the distinction between two types of HRS: type 1 and type 2. Before the introduction of the new therapeutic options, the median survival of patients with type 1 HRS was only 1.7 weeks, and 6-12 months in patients with type 2 HRS. Liver transplantation (LT) was the first therapeutic option to change the prognosis of cirrhotic patients with HRS and 5 year survival after LT in patients with HRS is only slightly less than that of transplanted patients without HRS and markedly increased when compared to survival in nontransplanted patients with HRS. Nevertheless, a large proportion of patients die before LT is possible because of the poor prognosis of HRS and the prolonged waiting times in most transplant centres. Other therapeutic approaches were therefore developed to increase survival in patients with HRS. Vasoconstrictors and transjugular intrahepatic portosystemic shunt (TIPS) are the most promising. The administration of vasoconstrictors together with albumin has been shown to reverse type 1 HRS and even to completely normalize renal function in 60-70% of treated patients. To date, four studies assessing TIPS in the management of type 1 HRS have been reported and TIPS insertion was technically successful in all of them. Given the shortage of donors for LT, vasoconstrictor therapy and TIPS strategies may be considered as a bridge to LT in patients with type 1 HRS. PMID- 15335402 TI - Review article: hepatorenal syndrome--how to assess response to treatment and nonpharmacological therapy. AB - Hepatorenal syndrome (HRS) is a complex syndrome. In addition to severe reduction of renal function due to renal vasoconstriction, there is impairment in systemic haemodynamics, activation of the renin-angiotensin and sympathetic nervous systems and antidiuretic hormone, vasoconstriction of the brain, muscle and skin, and dilutional hyponatraemia. Treatment in patients with type 2 HRS, the most frequent form of HRS, is directed towards managing refractory ascites. Paracentesis is the treatment of choice. TIPS is also effective but is more expensive, is associated with higher incidence of hepatic encephalopathy, and does not increase survival. Although a rapidly progressive renal failure is the most characteristic manifestation of type 1 HRS, there is failure in other organs such as the liver and the brain. A decrease in cardiac output develops in these patients, associated with a decrease in cardiopulmonary pressures. Since type 1 HRS mainly occurs in patients with spontaneous bacterial peritonitis and massive release of cytokines within the peritoneal cavity, it may be considered as a special form of multiorgan failure of circulatory origin. Not surprisingly, the treatment of choice in type 1 HRS is the combination of vasoconstrictors to reduce arterial vasodilation and plasma volume expansion with albumin to increase cardiac preload. TIPS is also effective in these patients and the combination of pharmacological treatment followed by TIPS may be the most effective approach. PMID- 15335404 TI - Review article: pharmacological treatment of hepatorenal syndrome. AB - Hepatorenal syndrome (HRS) is a common complication of advanced cirrhosis characterized not only by renal failure but also by marked alterations in systemic haemodynamics and activity of endogenous vasoactive systems. Renal failure is due to a severe vasoconstriction of the renal circulation. The pathogenesis of HRS is not completely understood but it is probably the result of extreme underfilling of the arterial circulation secondary to arterial vasodilation located in the splanchnic circulation. As well as the renal circulation, all other extrasplanchnic vascular beds appear to be vasoconstricted. The diagnosis of HRS is currently based on the exclusion of nonfunctional causes of renal failure; prognosis of patients with HRS is very poor. Liver transplantation is the best option in selected patients, but it is not always applicable as survival expectancy is short. Vasoconstrictor drugs with preferential effect on the splanchnic circulation (vasopressin analogues with a predominant V1 receptor effect, such as terlipressin--Glypressin) are very effective in improving renal function, with reversal of HRS being achieved in approximately two-thirds of patients. There is no agreement as to the terlipressin treatment regimen that is associated with a greater efficacy and lower incidence of side-effects. It appears that the administration of albumin together with terlipressin improves the therapeutic response rate. The impact of treatment on the natural course of HRS remains to be assessed in prospective investigations, but it seems that the reversal of HRS is associated with improved survival. Finally, treatment of patients with HRS with terlipressin before transplantation seems to improve post-transplantation outcome. PMID- 15335406 TI - Review article: future indications for terlipressin therapy. AB - Vasoconstrictor agents such as terlipressin (Glypressin) have been shown to have beneficial effects in the treatment of hepatorenal syndrome (HRS), in terms of improving renal function and subsequent survival rates. Patients with HRS have also been shown to have improved survival after liver transplantation if they receive terlipressin treatment prior to transplantation. In addition, studies show that terlipressin may have beneficial effects in treating other indications, including paracentesis-induced circulatory dysfunction and endotoxic shock. A positive effect has also been demonstrated with vasopressin in cardiopulmonary resuscitation. PMID- 15335408 TI - New pathophysiological mechanisms in irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is a functional, multifactorial disease characterized by abdominal pain and erratic bowel habit. Changes in gastrointestinal motor function, enhanced perception of stimuli arising from the gut wall and psychosocial factors are thought to be major contributors for symptom generation. In recent years, several additional factors have been identified and postulated to interact with these classical mechanisms. Reduced ability to expel intestinal gas with consequent gas trapping and bowel distension may contribute to abdominal discomfort/pain and bloating. Abnormal activation of certain brain regions following painful stimulation of the rectum suggests altered processing of afferent signals. An acute gastrointestinal infection is now a recognized aetiological factor for symptom development in a subset of IBS patients (i.e. post-infectious IBS), who are probably unable to down-regulate the initial inflammatory stimulus efficiently. Furthermore, low-grade inflammatory infiltration and activation of mast cells in proximity to nerves in the colonic mucosa may also participate in the frequency and severity of perceived abdominal pain in post-infectious and non-specific IBS. Initial evidence suggests the existence of changes in gut microflora, serotonin metabolism and a genetic contribution in IBS pathophysiology. These novel mechanisms may aid a better understanding of the complex pathophysiology of IBS and to develop new therapies. PMID- 15335409 TI - Diagnosis and therapy of irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is one of the most common gut functional diseases, affecting 10-20% of people worldwide. Although most patients do not seek medical help, the disease accounts for huge costs for both patients and health-care systems and worsens significantly patients' quality of life. Diagnosis is based on the identification of symptoms according to Manning, Rome I and Rome II criteria and exclusion of alarm indicators. IBS symptoms overlap with those of coeliac disease, lactose intolerance, food allergies and bile salt malabsorption. The treatment of IBS is centred on an excellent doctor-patient relationship along with drugs targeting the predominant symptom, especially during exacerbations. Current pharmacological remedies are unsatisfactory due to the high number of patients complaining of lack of response and/or symptom recurrence. Although useful in some IBS patients, the validity of psychotherapy deserves further investigation. A wide array of potentially useful drugs are currently under consideration in pre-clinical trials. A better understanding of the pathogenetic mechanisms underlying IBS may help to develop more effective drugs for this disease. PMID- 15335410 TI - What the physician needs to know for correct management of gastro-oesophageal reflux disease and dyspepsia. AB - Upper gastrointestinal symptoms are highly prevalent; usually those consulting have multiple symptoms, confounding management. Here, common clinically relevant management issues are considered based on the best available evidence. Regardless of the presenting symptoms, determine if there are any alarm features; these have a low positive predictive value for malignancy but all patients with them should be referred for prompt upper gastrointestinal endoscopy. Ask about medications; of most importance are the non-steroidal anti-inflammatory drugs (NSAIDs), both non-selective and COX-2 selective. Try to ascertain if the symptom pattern suggests gastro-oesophageal reflux disease (GERD) or not. Dominant heartburn, however, may be of limited value; if the background prevalence of GERD is 25% and the patient complains of dominant heartburn, then the likelihood that such a patient has GERD as identified by 24-h oesophageal pH testing is only just over 50%. If reflux disease is strongly suspected and there are no alarm features, give an empirical trial of a proton pump inhibitor (PPI). Symptoms cannot separate adequately functional from organic dyspepsia. Endoscopy in dyspepsia with no alarm features is more costly than an empirical management approach. H. pylori testing and treatment remains in most settings the preferable initial choice for managing dyspepsia without obvious GERD. However, a PPI trial may offer a similar outcome and may be preferable in low H. pylori prevalence areas; head-to-head management trials in primary care are lacking. PMID- 15335411 TI - Helicobacter pylori infection: anything new should we know? AB - Over the past year, 2003-4, there have been a number of studies consolidating previous work in relation to pathogenesis of disease, diagnosis and management of Helicobacter pylori. Studies into the pathogenesis of disease have identified the main adhesin of H. pylori as an important virulence marker and as a potential target for therapy. Molecular investigations of both the strain and host variations have identified the action of several of the virulence factors, e.g. cagA, vacA, on disrupting host cell signalling and the consequences in respect of the release of chemokines from the damaged gastric epithelium and the effect on apoptosis. Over the past year, there have been further diagnostic kits developed based on modifications of current technology. Two promising areas of research for diagnosis are the use of host/strain genome polymorphisms as a means of identifying high-risk patients who may develop severe disease and the use of proteomics to identify potential antigens of diagnostic (or therapeutic) use. The three main antibiotics that are used in first-line eradication regimens are clarithromycin, metronidazole and amoxycillin. Of these, metronidazole has the highest prevalence of resistance, followed by clarithromycin; amoxycillin resistance is only rarely reported. The decreasing success of current first-line therapy is the driving force for the development of new antibiotic combinations and a search for novel sources for chemotherapeutic agents and novel therapeutic targets. PMID- 15335412 TI - Screening for gastric cancer: does it make sense? AB - Gastric cancer remains the second most frequent and lethal malignancy worldwide, with a wide range of incidence rates in different populations. Reducing its incidence is a priority in many high-risk countries, and screening programmes are often advocated as an option. An effective screening programme should have the following characteristics: (i) the disease should be common in the population, otherwise the individual benefit will not offset the risk, cost and inconvenience of screening of the rest of the population; (ii) the diagnostic test(s) used should be safe, simple, inexpensive and reliable; and (iii) effective treatment should be available. With respect to gastric cancer, these conditions are not fulfilled completely in any population and therefore population-based screening does not appear to be a viable approach to the prevention of this neoplasia. In contrast, gastroscopic screening in selected high-risk subjects would seem appropriate. A more effective strategy, particularly in high-incidence populations, could be the widespread screening and treatment for Helicobacter pylori infection. This could result in the virtually complete elimination of chronic gastritis and its associated conditions, including most peptic ulcers, primary gastric B-cell lymphomas and a major portion of gastric epithelial tumours. PMID- 15335413 TI - Gastrointestinal side-effects of traditional non-steroidal anti-inflammatory drugs and new formulations. AB - Although adverse effects of non-steroidal anti-inflammatory drugs (NSAIDs) occur in only a small proportion of users, the widespread use of these drugs has resulted in a substantial overall number of affected persons who experience serious gastrointestinal complications. Dyspeptic symptoms are estimated to occur in 10-60% of NSAID users and lead to discontinuation of treatment in 5-15% of rheumatoid arthritis patients taking NSAIDs. It is now well established that the point prevalence of peptic ulcer disease in patients receiving conventional NSAID therapy ranges between 10 and 30%, representing a 10-30-fold increase over that found in the general population. One of 175 users of conventional NSAIDs in the USA will be hospitalized each year for NSAID-induced gastrointestinal damage. The mortality of hospitalized patients remains about 5-10%, with an expected annual death rate of 0.08%. The selective COX-II inhibitors (rofecoxib, celecoxib, parecoxib, etoricoxib, valdecoxib, lumiracoxib) show consistently comparable efficacy to that of conventional non-steroidal anti-inflammatory drugs (NSAIDs) in patients with rheumatoid arthritis and osteoarthritis, but have a significantly reduced propensity to cause gastrointestinal toxicity. In many cases, the gastric effects of therapeutically active doses of COX-II inhibitors are indistinguishable from placebo. The safety benefits of COX-2 inhibitors given alone appear similar to combined therapy with conventional NSAIDs and gastroprotective agents. These findings warrant the consideration of COX-II inhibitors as first-line therapy in patients requiring long-term pain control. PMID- 15335414 TI - Non-steroidal anti-inflammatory drugs: who should receive prophylaxis? AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are the major recognized cause of iatrogenic disease, and may cause 100 000 deaths per annum through peptic ulcer complications. A number of risk factors can be identified that indicate patients at high risk. These patients can be managed by substitution of a COX-2 inhibitor or by prophylaxis with a proton pump inhibitor (PPI). Because risk factors that render patients at high risk of ulcer complications also act in the absence of NSAID use, PPI prophylaxis (or Helicobacter pylori eradication where H. pylori is the risk factor) have much to offer and controlled studies show that the incidence of recurrent peptic ulcer bleeding can be reduced substantially by PPI co-administration. Substitution of COX-2 inhibitors also has much to offer, arguably most in those without risk factors (although regulatory authorities do not accept this argument). Recent data show that PPI and COX-2 inhibitors can play complementary roles in the management of patients with moderate to severe dyspepsia and at high risk of ulcer complications. PMID- 15335415 TI - Should we eradicate Helicobacter pylori in non-steroidal anti-inflammatory drug users? AB - The interaction between Helicobacter pylori and non-steroidal anti-inflammatory drugs (NSAIDs) in ulcerogenesis has been visited by many studies. Apparently these studies yielded conflicting results. This is a result of a wide diversity of methodology, selection of patient groups and definitions of outcome used by different investigators. This review attempts to analyse separately studies dealing with new or chronic NSAID users, primary or secondary prophylaxis, complicated or uncomplicated ulcers in NSAID or aspirin users. Evidence suggests that eradication of Helicobacter pylori infection may reduce the risk of ulcer and ulcer complications in patients requiring NSAIDs and aspirin. Whether or not one should test-and-treat H. pylori before prescribing NSAIDs is a complicated issue. Factors such as the ulcer risk of patients, previous history of NSAID usage and the use of aspirin or NSAIDs would guide the strategy. PMID- 15335417 TI - Experimental radiobiological investigations into radiosurgery: present understanding and future directions. AB - LARS LEKSELL BEGAN radiobiological investigations to study the effect of high dose focused radiation on the central nervous system more than 5 decades ago. Although the effects of radiosurgery on the brain tumor microenvironment are still under investigation, radiosurgery has become a preferred management modality for many intracranial tumors and vascular malformations. The effects and the pathogenesis of biological effects after radiosurgery may be unique. The need for basic research concerning the radiobiological effects of high-dose, single fraction, ionizing radiation on nervous system tissue is crucial. Information from those studies would be useful in devising strategies to avoid, prevent, or ameliorate damage to normal tissue without compromising treatment efficacy. The development of future applications of radiosurgery will depend on an increase in our understanding of the radiobiology of radiosurgery, which in turn will affect the efficacy of treatment. This article analyzes the current state of radiosurgery research with regard to the nature of central nervous system effects, the techniques developed to increase therapeutic efficacy, investigations into the use of radiosurgery for functional disorders, radiosurgery as a tool for investigations into basic central nervous system biology, and the additional areas that require further investigation. PMID- 15335418 TI - Changing neurosurgical workload in the United States, 1988-2001: craniotomy other than trauma in adults. AB - OBJECTIVE: Changes in neurosurgical workload can justify requests for hospital resources and guide planning by neurosurgical training programs. Most previous studies have used non-population-based data sources, such as surveys of professional society members, to explore the neurosurgical workload in the United States. METHODS: This is a retrospective cohort study of patients in Diagnosis Related Group (DRG) 1 ("Craniotomy other than trauma, age > 17") using the Nationwide Inpatient Sample. Statistical methods were adjusted for complex survey methodology to generate total United States caseload estimates. RESULTS: The total United States DRG 1 caseload increased from 70,800 admissions in 1988 to 105,300 admissions in 2001, a 50% relative increase (P < 0.001). For most diagnostic categories, the relative caseload increase was similar to that for the whole group. Patient age and sex distributions remained stable over time. Medical comorbidities, such as hypertension, chronic pulmonary disease, diabetes, and obesity, became more frequent. Elective admissions increased and in-hospital mortality rates decreased. Length of hospital stay decreased during the first half of the study period and then stabilized. Combined with increasing caseload, this caused total annual inpatient DRG 1 days to increase progressively after 1996. The number of United States hospitals with DRG 1 admissions decreased over time. Per-hospital annual DRG 1 caseloads increased, especially at high-volume centers. For the largest 100 hospitals by DRG 1 caseload, total admissions increased from 8.5% of all United States admissions (1988) to 9.4% (2001), whereas DRG 1 caseload increased disproportionately, from 27% to 38% of the United States aggregate caseload. This is evidence that progressive centralization of DRG 1 admissions took place during the study period. CONCLUSION: We documented an increase in total caseload and centralization of care for DRG 1 in the United States during the period 1988 to 2001. Defining the reasons for the changes in neurosurgical workload we observed will require further research. PMID- 15335419 TI - Fractionated stereotactic radiotherapy for the treatment of large arteriovenous malformations with or without previous partial embolization. AB - OBJECTIVE: Despite the success of stereotactic radiosurgery, large inoperable arteriovenous malformations (AVMs) of 14 cm(3) or more have remained largely refractory to stereotactic radiosurgery, with much lower obliteration rates. We review treatment of large AVMs either previously untreated or partially obliterated by embolization with fractionated stereotactic radiotherapy (FSR) regimens using a dedicated linear accelerator (LINAC). METHODS: Before treatment, all patients were discussed at a multidisciplinary radiosurgery board and found to be suitable for FSR. All patients were evaluated for pre-embolization. Those who had feeding pedicles amenable to glue embolization were treated. LINAC technique involved acquisition of a stereotactic angiogram in a relocatable frame that was also used for head localization during treatment. The FSR technique involved the use of six 7-Gy fractions delivered on alternate days over a 2-week period, and this was subsequently dropped to 5-Gy fractions after late complications in one of seven patients treated with 7-Gy fractions. Treatments were based exclusively on digitized biplanar stereotactic angiographic data. We used a Varian 600SR LINAC (Varian Medical Systems, Inc., Palo Alto, CA) and XKnife treatment planning software (Radionics, Inc., Burlington, MA). In most cases, one isocenter was used, and conformality was established by non-coplanar arc beam shaping and differential beam weighting. RESULTS: Thirty patients with large AVMs were treated between January 1995 and August 1998. Seven patients were treated with 42-Gy/7-Gy fractions, with one patient lost to follow-up and the remaining six with previous partial embolization. Twenty-three patients were treated with 30-Gy/5-Gy fractions, with two patients lost to follow-up and three who died as a result of unrelated causes. Of 18 evaluable patients, 8 had previous partial embolization. Mean AVM volumes at FSR treatment were 23.8 and 14.5 cm(3), respectively, for the 42-Gy/7-Gy fraction and 30-Gy/5-Gy fraction groups. After embolization, 18 patients still had AVM niduses of 14 cm(3) or more: 6 in the 7-Gy cohort and 12 in the 5-Gy cohort. For patients with at least 5-year follow-up, angiographically documented AVM obliteration rates were 83% for the 42-Gy/7-Gy fraction group, with a mean latency of 108 weeks (5 of 6 evaluable patients), and 22% for the 30-Gy/5-Gy fraction group, with an average latency of 191 weeks (4 of 18 evaluable patients) (P = 0.018). For AVMs that remained at 14 cm(3) or more after embolization (5 of 6 patients), the obliteration rate remained 80% (4 of 5 patients) for the 7-Gy cohort and dropped to 9% for the 5-Gy cohort. A cumulative hazard plot revealed a 7.2-fold greater likelihood of obliteration with the 42-Gy/7-Gy fraction protocol (P = 0.0001), which increased to a 17-fold greater likelihood for postembolization AVMs of 14 cm(3) or more (P = 0.003). CONCLUSION: FSR achieves obliteration for AVMs at a threshold dose, including large residual niduses after embolization. With significant treatment related morbidities, further investigation warrants a need for better three dimensional target definition with higher dose conformality. PMID- 15335420 TI - Cohort study of intraventricular thrombolysis with recombinant tissue plasminogen activator for aneurysmal intraventricular hemorrhage. AB - OBJECTIVE: Thrombolytic agents have been administered through external ventricular drains to treat intraventricular hemorrhage, the goals being to accelerate clot clearance, prevent catheter obstruction, and help control intracranial pressure. We compared these variables in a group of aneurysm patients treated by one surgeon who routinely used intraventricular recombinant tissue plasminogen activator (rt-PA) for obstructive hematocephalus with those in a group of similar patients treated by other surgeons who did not. METHODS: Patients included in this analysis were those with repaired cerebral aneurysms causing hemorrhage into at least three ventricles with ventriculomegaly requiring external ventricular drainage. The ventricular system was considered "opened" when all ventricles were patent and reduced in size on computed tomographic scans. Those treated with rt-PA received 4 mg/d through a ventricular drain until ventricular opening. RESULTS: The mean number of days to ventricular opening was 3.9 (standard deviation [SD], 1.0) for the 21 patients treated with rt-PA and 7.1 (SD, 3.7) for the 9 who were not (P = 0.001), and the mean intracranial pressure (mm Hg) 24 hours after treatment with rt-PA was 10.4 (SD, 6.1) compared with 14.1 (SD, 5.9) during the same interval for the group that did not receive rt-PA (P = 0.13). Ventricular catheter replacement was required in 1 rt-PA patient (for a misplaced catheter, before rt-PA treatment) and 3 patients who did not receive rt PA (all for catheter obstructions with blood clot) (P = 0.07), and ventriculoperitoneal shunts were placed in 4 rt-PA patients and 3 patients who did not receive rt-PA (P = 0.4). CONCLUSION: Intraventricular thrombolysis with rt-PA seems to assist in the acute management of patients with large aneurysmal intraventricular hemorrhages, speeding clearance of aneurysmal intraventricular hemorrhage, normalizing intracranial pressure, and reducing ventricular catheter obstruction. A randomized trial is needed to confirm these findings, establish treatment safety, and determine whether treatment affects outcome. PMID- 15335421 TI - Variations on the standard transsphenoidal approach to the sellar region, with emphasis on the extended approaches and parasellar approaches: surgical experience in 105 cases. AB - OBJECTIVE: The traditional boundaries of the transsphenoidal approach may be expanded to include the region from the cribriform plate of the anterior cranial base to the inferior clivus in the anteroposterior plane, and laterally to expose the cavernous cranial nerves and the optic canal. We review our combined experience with these variations on the transsphenoidal approach to various lesions of the sellar and parasellar region. METHODS: From 1982 to 2003, we used the extended and parasellar transsphenoidal approaches in 105 patients presenting with a variety of lesions of the parasellar region. This study specifically reviews the breadth of pathological lesions operated and the complications associated with the approaches. RESULTS: Variations of the standard transsphenoidal approach have been used in the following series: 30 cases of pituitary adenomas extending laterally to involve the cavernous sinus, 27 craniopharyngiomas, 11 tuberculum/diaphragma sellae meningiomas, 10 sphenoid sinus mucoceles, 18 clivus chordomas, 4 cases of carcinoma of the sphenoid sinus, 2 cases of breast carcinoma metastatic to the sella, and 3 cases of monostotic fibrous dysplasia involving the clivus. There was no mortality in the series. Permanent neurological complications included one case of monocular blindness, one case of permanent diabetes insipidus, and two permanent cavernous cranial neuropathies. There were four cases of internal carotid artery hemorrhage, one of which required ligation of the cervical internal carotid artery and resulted in hemiparesis. The incidence of postoperative cerebrospinal fluid fistulae was 6% (6 of 105 cases). CONCLUSION: These modifications of the standard transsphenoidal approach are useful for lesions within the boundaries noted above, they offer excellent alternatives to transcranial approaches for these lesions, and they avoid prolonged exposure time and brain retraction. Technical details are discussed and illustrative cases presented. PMID- 15335422 TI - On-line flow cytometry for real-time surgical guidance. AB - OBJECTIVE: This study tests the feasibility of using on-line analysis of tissue during surgical resection of brain tumors to provide biologically relevant information in a clinically relevant time frame to augment surgical decision making. For the purposes of establishing feasibility, we used measurement of deoxyribonucleic acid (DNA) content as the end point for analysis. METHODS: We investigated the feasibility of interfacing an ultrasonic aspiration (USA) system with a flow cytometer (FC) capable of analyzing DNA content (DNA-FC). The sampling system design, tissue preparation requirements, and time requirements for each step of the on-line analysis system were determined using fresh beef brain tissue samples. We also compared DNA-FC measurements in 28 nonneoplastic human brain samples with DNA-FC measurements in specimens of 11 glioma patients obtained from central tumor regions and surgical margins after macroscopically gross total tumor removal to estimate the potential for analysis of a biological marker to influence surgical decision making. RESULTS: With minimal modification, modern FC systems are fully capable of real-time, intraoperative analysis of USA specimens. The total time required for on-line analysis of USA specimens varies between 36 and 63 seconds; this time includes delivery from the tip of the USA to complete analysis of the specimen. Approximately 60% of this time is required for equilibration of the DNA stain. When compared with values for nonneoplastic human brain samples, 50% of samples (10 of 20) from macroscopically normal glioma surgical margins contained DNA-FC abnormalities potentially indicating residual tumor. CONCLUSION: With an interface of existing technologies, DNA content of brain tissue samples can be analyzed in a meaningful time frame that has the potential to provide real-time information for surgical guidance. The identification of DNA content abnormalities in macroscopically normal tumor resection margins by DNA-FC supports the practical potential for on-line analysis of a tumor marker to guide surgical resections. The development of such a device would provide neurosurgeons with an objective method for intraoperative analysis of a clinically relevant biological parameter that can be measured in real time. PMID- 15335423 TI - Angiogenic factors in the cerebrospinal fluid of patients with astrocytic brain tumors. AB - OBJECTIVE: Gliomas account for most primary brain tumors in adults, and survival correlates with the grade and vascularity of the tumor. The degree of tumor related angiogenesis seems to be a significant predictor of tumor progression, recurrence, and metastatic spread in a variety of malignant diseases, including brain tumors. Our study's objective was to quantify the levels of two angiogenic factors, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), in the cerebrospinal fluid (CSF) and serum of patients with gliomas and to correlate these levels with tumor grade, vascularity, and overall survival. METHODS: Twenty-six patients with the diagnosis of cerebral glioma (19 high-grade, 7 low-grade) comprised the study group. Ten patients with communicating hydrocephalus served as controls. Levels of VEGF and bFGF in the CSF and serum were determined using enzyme-linked immunosorbent assay analysis. Tumor vascularity was graded qualitatively using immunohistochemical staining for CD34. Nonparametric statistical techniques were used for data analysis. RESULTS: Median levels of bFGF and VEGF in the CSF were significantly higher in patients with high-grade glioma as compared with patients with low-grade glioma or hydrocephalus (bFGF levels, 52, 26, and 24 ng/ml, respectively, P < 0.0001; VEGF levels, 17.6, 7.2, and 8.3 ng/ml, respectively, P < 0.005). A significant correlation was found comparing CSF levels of bFGF with levels of VEGF (P < 0.001). The levels of the angiogenic factors in the CSF correlated with the degree of tumor vascularity and were adversely associated with patient survival. Serum levels of the angiogenic factors showed no correlation to tumor grade, vascularity, or survival. CONCLUSION: Our data suggest that CSF levels of bFGF and VEGF may serve as an additional marker for tumor grading and vascularity and may help predict survival. PMID- 15335424 TI - Lesion-induced pseudo-dominance at functional magnetic resonance imaging: implications for preoperative assessments. AB - OBJECTIVE: To illustrate how lesion-induced neurovascular uncoupling at functional magnetic resonance imaging (fMRI) can mimic hemispheric dominance opposite the side of a lesion preoperatively. METHODS: We retrospectively reviewed preoperative fMRI mapping data from 50 patients with focal brain abnormalities to establish patterns of hemispheric dominance of language, speech, visual, or motor system functions. Abnormalities included gliomas (31 patients), arteriovenous malformations (AVMs) (11 patients), other congenital lesions (4 patients), encephalomalacia (3 patients), and tumefactive encephalitis (1 patient). A laterality ratio of fMRI hemispheric dominance was compared with actual hemispheric dominance as verified by electrocortical stimulation, Wada testing, postoperative and posttreatment deficits, and/or lesion-induced deficits. fMRI activation maps were generated with cross-correlation (P < 0.001) or t test (P < 0.001) analysis. RESULTS: In 50 patients, a total of 85 functional areas were within 5 mm of the edge of a potentially resectable lesion. In 23 of these areas (27%), reduced fMRI signal in perilesional eloquent cortex in conjunction with preserved or increased signal in homologous contralateral brain areas revealed functional dominance opposite the side of the lesion. This suggested possible lesion-induced transhemispheric cortical reorganization to homologous brain regions (homotopic reorganization). In seven patients, however, the fMRI data were inconsistent with other methods of functional localization. In two patients with left inferior frontal gyrus gliomas and in one patient with focal tumefactive meningoencephalitis, fMRI incorrectly suggested strong right hemispheric speech dominance. In two patients with lateral precentral gyrus region gliomas and one patient with a left central sulcus AVM, the fMRI pattern incorrectly suggested primary corticobulbar motor dominance contralateral to the side of the lesion. In a patient with a right superior frontal gyrus AVM, fMRI revealed pronounced left dominant supplementary motor area activity in response to a bilateral complex motor task, but right superior frontal gyrus perilesional hemorrhage and edema subsequently caused left upper-extremity plegia. Pathophysiological factors that might have caused neurovascular uncoupling and facilitated pseudo-dominance at fMRI in these patients included direct tumor infiltration, neovascularity, cerebrovascular inflammation, and AVM-induced hemodynamic effects. Sixteen patients had proven (1 patient), probable (2 patients), or possible (13 patients) but unproven lesion-induced homotopic cortical reorganization. CONCLUSION: Lesion-induced neurovascular uncoupling causing reduced fMRI signal in perilesional eloquent cortex, in conjunction with normal or increased activity in homologous brain regions, may simulate hemispheric dominance and lesion-induced homotopic cortical reorganization. PMID- 15335425 TI - Decreases in ventricular volume correlate with decreases in ventricular pressure in idiopathic normal pressure hydrocephalus patients who experienced clinical improvement after implantation with adjustable valve shunts. AB - OBJECTIVE: This retrospective study examined whether changes in ventricular volume correspond with changes in adjustable valve pressure settings in a cohort of patients who received shunts to treat idiopathic normal pressure hydrocephalus. We also examined whether these pressure-volume curves and other patient variables would co-occur with a positive clinical response to shunting. METHODS: We selected 51 patients diagnosed with idiopathic normal pressure hydrocephalus who had undergone implantation of a Codman Hakim programmable valve (Medos S.A., Le Locle, Switzerland). Clinical data were gathered from the patients' records and clinical notes by an investigator blinded to patients' ventricular volumes. Ventricular volume was measured using 3D Slicer, an image analysis and interactive visualization software package developed and maintained at the Surgical Planning Laboratory at Brigham and Women's Hospital. RESULTS: Eighty-six percent of patients with gait disturbance at presentation showed improvement of this symptom, 70% experienced improvement in incontinence, and 69% experienced improvement in dementia. For the group showing 100% clinical improvement, the correlation coefficient of average changes in valve pressure over time (delta P/delta T) and average changes in ventricular volume over time (delta V/delta T) were high at 0.843 (P < 0.05). For the group experiencing no or only partial improvement, the correlation coefficient was 0.257 (P = 0.32), indicating no correlation between average delta V/delta T and average delta P/delta T for each patient. CONCLUSION: This was a carefully analyzed modeling study of idiopathic normal pressure hydrocephalus treatment made possible only by adjustable valve technology. With careful volumetric analysis, we found that changes in ventricular volume correlated with adjustments in valve pressure settings for those patients who improved clinically after shunting. This suggests that positive clinical responders retained parenchymal elasticity, emphasizing the importance of dynamic changes in this cohort. PMID- 15335426 TI - Risk of infection with prolonged ventricular catheterization. AB - OBJECTIVE: The relationship between extended ventricular catheterization and infection remains controversial. Although studies have substantiated an increasing infection rate with prolonged catheterization, there has been less agreement on whether this trend continues beyond 10 days. Our study reviews the daily infection rate of 595 patients, 213 of whom underwent more than 10 days of catheterization. METHODS: All patients who underwent ventricular monitoring in the neurological intensive care unit from 1995 to 2003 at the University of Michigan Health System were reviewed retrospectively. Infection was defined as a positive cerebrospinal fluid culture. Life-table analysis was used to calculate daily hazard (infection) rates. Patient age, sex, diagnosis, catheter exchanges, location of patient during catheter insertion, and cerebrospinal fluid leak were evaluated as risk factors for infection. RESULTS: The average patient age was 51.3 years, and 51.3% were male. Duration of catheterization averaged 8.6 days. The overall infection rate was 8.6%. Daily infection rates increased from the onset of catheter insertion but reached a plateau after Day 4, with subsequent rates ranging predominantly between 1 and 2%, even with extended catheterization beyond 10 days. Only ventricular catheters that had been placed at other institutions significantly affected the infection rate. CONCLUSION: A relationship between duration of catheterization and infection seems to be present. However, this relationship is not linear. There is an extremely low daily infection rate that rises over the initial 4 days but then remains relatively constant even with prolonged catheter use. Clinical decisions to continue ventricular catheterization should reflect this low daily risk of infection, which does not seem to increase with extended catheter use. PMID- 15335427 TI - Craniocerebral aspergillosis of sinonasal origin in immunocompetent patients: clinical spectrum and outcome in 25 cases. AB - OBJECTIVE: Craniocerebral aspergillosis of sinonasal origin has been reported mainly in immunocompromised patients with high mortality, and it has been described very infrequently in immunocompetent hosts. This retrospective study focuses on clinical outcome in relation to anatomic locations of invasive aspergillosis of sinonasal origin in immunocompetent patients with emphasis on our preliminary experience with use of preoperative orally administered itraconazole. METHODS: Medical records of patients treated in two tertiary care hospitals from 1991 to 2003 were reviewed retrospectively. All patients had radiological evidence of disease in the paranasal sinuses with or without intracranial extension. The study cohort was divided into three types on the basis of area of involvement revealed by computed tomographic or magnetic resonance imaging scans of brain. All patients underwent surgical intervention and treatment with antifungal therapy. Preoperative orally administered itraconazole therapy was used in four patients on the basis of neuroradiological features. Clinical outcome was assessed with the Glasgow Outcome Scale, and univariate analysis of prognostic factors was performed with 95% confidence interval (P = 0.05). RESULTS: Mean patient age was 36.5 years (range, 14-74 yr) with a male preponderance (male-to-female ratio, 23:2). Nasal stuffiness (n = 13), headaches (n = 10), proptosis (n = 9), and nasal discharges (n = 7) were major presenting clinical features. Radiological data were obtained by computed tomographic (n = 25) and magnetic resonance imaging (n = 20) scans of the brain, and diagnoses were established by histopathological analysis (n = 20) or/and fungal cultures (n = 15). Preoperative orally administered itraconazole was given in four patients with intracerebral aspergillosis. Overall mortality was 28% and was highest in patients with Type 1 aspergillosis (66.7%). Type 3 aspergillosis and use of preoperative itraconazole remained statistically significant prognostic factors. CONCLUSION: Craniocerebral aspergillosis in immunocompetent hosts has three patterns of presentation that seem to correlate with clinical outcomes. Intracerebral aspergillosis (Type 1) is associated with the worst clinical outcome. Patients with orbital and cranial base aspergillosis (Type 3) had good recovery. Intracranial extradural aspergillosis (Type 2) remained intermediate on the Glasgow Outcome Scale. Preoperative orally administered itraconazole therapy may improve clinical outcome in patients with intracerebral aspergillosis. Prospective clinical studies are required to make firm clinical therapeutic recommendations. PMID- 15335428 TI - Atlantoaxial rotatory fixation: Part 1--Biomechanics of normal rotation at the atlantoaxial joint in children. AB - OBJECTIVE: Atlantoaxial rotatory fixation (AARF) remains a recondite entity loosely included under the panoply of cervical trauma. The difficulty in finding a precise definition and reliable diagnostic criteria for AARF has been chiefly because of a lack of normative biomechanical data for C1-C2 rotation. As Part 1 and foundation of a comprehensive undertaking to define the biomechanics, mechanism, diagnosis, classification, and management of AARF, the present study focuses on the dynamic behavior of C1 and C2 during normal voluntary head rotation in children. METHODS: Twenty-one normal children 3 to 11.5 years old underwent computed tomographic examinations from the lower clivus to the base of C3 in various head positions during axial rotation. The angles made by C1, C2, and the occiput with the vertical 0 degrees were recorded, and from these, the separation angles between C1 and C2 (C1-C2 degrees) were calculated for each head position (represented by the C1 angle) studied. In 18 children, the range of rotation was between 90 and -90 degrees, i.e., with the head making a full 180 degree turn from one side to the other. In 3 children, the head was first turned from 0 to 90 degrees and then back from 90 to 0 degrees, making only a half turn. All separation angles (C1-C2 degrees) were then plotted against the corresponding C1 angle to create a motion curve, which, in essence, describes the interaction between C1 and C2 through the full range of head positions. In the 18 children with full turns, both individual motion curves and a composite motion curve comprising all data were constructed. RESULTS: There is a high degree of concordance for rotational behavior of C1 and C2 in the 18 subjects undergoing full turn. C1 always crosses C2 at or near 0 degrees, the null point of full rotation. The predictable relationship between C1 and C2 is depicted by three distinct regions on the composite motion curve: when C1 rotates from 0 to 23 degrees, it moves alone, with C2 remaining stationary at approximately 0 degrees (the single-motion phase). When C1 rotates from 24 to 65 degrees, C1 and C2 move together, but C1 always moves at a faster rate (the double-motion phase), C2 being pulled by yoking ligaments. From 65 degrees onward, C1 and C2 move in exact unison (the unison-motion phase) with a fixed, maximum separation angle of approximately 43 degrees, head rotation being carried exclusively by the subaxial segments. In the 3 children with half turn, the forward rotation curve and the reverse rotation curve are almost superimposable, suggesting that the "yoking" between C1 and C2 is a result of more than just tensing and relaxing of ligaments but probably also to a mutual dragging by irregular bony surfaces between the two bones. CONCLUSION: C1 and C2 in children move in a predictable manner during axial head rotation, with a high degree of concordance among subjects and a relatively narrow variance from the mean. The composite motion curve can thus be used as a touchstone against which may be judged all manners of pathological interlock or "stickiness" between C1 and C2 in rotation that could be defined as AARF. PMID- 15335429 TI - Effect of vagus nerve stimulator magnet on programmable shunt settings. AB - OBJECTIVE: Vagus nerve stimulators and programmable shunt valves are used in the operative care of epilepsy and hydrocephalus, respectively. Both devices use magnetic fields to activate and program their various settings and functions. The authors conducted several ex vivo trials to better elucidate any interplay between the two systems. METHODS: A pulse generator controller (Cyberonics Corp., Houston, TX) was brought to within 4 cm of Strata programmable shunt valves (Medtronic Neurosurgery, Goleta, CA). Each of five valves was preset to either a low- or high-pressure setting and then challenged with the vagus nerve stimulator generator. Each valve was challenged 20 times, for a total of 100 trials. RESULTS: In 100 trials, 78 inadvertent pressure setting adjustments were recorded. In 46 attempts, the valve pressure was increased, and in 34 attempts, the pressure was decreased. CONCLUSION: This study provides some support to the anecdotal reports of inadvertent adjustments of programmable shunt valves by the external magnetic field created by vagus nerve stimulator pulse generator controllers. Further trials and a double-blind study are necessary to illustrate more clearly the possible relationship of these magnetically controlled neurosurgical devices. PMID- 15335430 TI - Treatment of extracranial and intracranial aneurysms and arteriovenous fistulae using stent grafts. AB - OBJECTIVE: Treatment of 11 patients with aneurysms or arteriovenous fistulae of the craniocervical arteries with stent grafts. METHODS: Peripheral stent grafts were deployed in two extracranial internal carotid arteries. Coronary stent grafts were used to treat two giant aneurysms, five direct carotid-cavernous fistulae, one vertebrojugular fistula, and two dissecting aneurysms of the vertebral artery (V2 and V4). RESULTS: Stent grafts were used successfully in two extracranial internal carotid and two extracranial vertebral arteries, one dissecting aneurysm of the intracranial vertebral artery, one giant aneurysm and one pseudoaneurysm of the cavernous internal carotid artery, and five direct carotid-cavernous sinus fistulae. Angiographic follow-up examinations (available in nine patients; obtained at 3 mo to 5 yr; average, 24 mo) revealed normal vessel caliber, and the stent grafts in all 9 of 11 initial patients were patent. There was a recurrent saccular aneurysm adjacent to the stent graft in the patient with the intracranial vertebral artery aneurysm. The following five complications were encountered: transient hemiparesis (n = 2), increased hemiparesis, postprocedural management-related fatality, and ICA dissection. In six patients, stent graft deployment was accomplished without any technical or clinical complication. There were no permanent neurological deficits consequent to stent graft placement. CONCLUSION: Stent grafts are a useful tool for the endovascular treatment of head and neck aneurysms and direct arteriovenous fistulae in selected patients. The major disadvantage of the currently available stent grafts is their lack of mechanical flexibility. Maneuvering stent grafts in the intracranial arteries carries the risk of iatrogenic vessel dissection and may require supportive measures and protection of the target site by conventional stents. PMID- 15335431 TI - Regeneration of peripheral nerve gaps with a polyglycolic acid-collagen tube. AB - OBJECTIVE: The aim of this study was to report by means of objective methods on the effectiveness of a nerve reconstruction procedure using a bioresorbable tube in two patients. Our previous successes in regenerating canine peripheral nerves across long distances (80-mm gaps) using a bioabsorbable tube have led us to investigate the value of such a tube for the treatment of human patients with chronic nerve injuries. METHODS: The device was made from a cylindrically woven polyglycolic acid tube filled with a collagen sponge. It was designed to be resorbed after nerve regeneration. Peripheral sensory nerve defects in two patients with neuroma and pain were reconstructed using this tube. Patient 1 (a 62-year-old man) had a 20-mm defect of the proper digital nerve, and Patient 2 (a 56-year-old woman) had a 65-mm defect of the superficial peroneal nerve. RESULTS: After surgery, both patients recovered from the unpleasant sensations and intolerable pain. In Patient 1, functional recovery was objectively identified at 2 months, and conduction velocity of the nerve recovered to 49.1 m/s. In Patient 2, conduction velocity of the nerve was determined to be 16.9 m/s at 5 months. Current perception threshold testing indicated that sensory nerve function had been recovered by 65 days after surgery. CONCLUSION: This work represents the first precise clinical evaluation, performed under objective evaluation criteria, of sensory recovery achieved using a nerve tube, suggesting that the use of a polyglycolic acid-collagen tube has the potential to become a viable alternative to conventional autografting for the repair of peripheral nerve defects. PMID- 15335432 TI - Analysis and evolution of head injury in football. AB - OBJECTIVE: To review head injury in football through historical, anatomic, and physiological analysis. METHODS: We obtained data from a thorough review of the literature. RESULTS: The reported incidence of concussion among high school football players dropped from 19% in 1983 to 4% in 1999. During the 1997 Canadian Football League season, players with a previous loss of consciousness in football were 6.15 times more likely to experience a concussion than players without a previous loss of consciousness (P < 0.05). Players with a previous concussion in football were 5.10 times more likely to experience a concussion than players without a previous concussion (P = 0.0001). With the implementation of National Operating Committee on Standards for Athletic Equipment standards, fatalities decreased by 74% and serious head injuries decreased from 4.25 per 100,000 to 0.68 per 100,000. CONCLUSION: Significant declines in both the incidence and severity of head injury have been observed. The enhanced safety records in football can be attributed to the application of more stringent tackling regulations as well as the evolving football helmet. The role of a neurosurgeon is critical in further head injury prevention and guidelines in sport. PMID- 15335433 TI - Birth and evolution of the football helmet. AB - OBJECTIVE: To review the advent and evolution of the football helmet through historical, physiological, and biomechanical analysis. METHODS: We obtained data from a thorough review of the literature. RESULTS: Significant correlation exists between head injuries and the advent of the football helmet in 1896, through its evolution in the early to mid-1900s, and regulatory standards for both helmet use and design and tackling rules and regulations. With the implementation of National Operating Committee on Standards for Athletic Equipment standards, fatalities decreased by 74% and serious head injuries decreased from 4.25 per 100,000 to 0.68 per 100,000. Not only is the material used important, but the protective design also proves essential in head injury prevention. Competition among leading helmet manufacturers has benefited the ultimate goal of injury prevention. However, just as significant in decreasing the incidence and severity of head injury is the implementation of newer rules and regulations in teaching, coaching, and governing tackling techniques. CONCLUSION: Helmet use in conjunction with more stringent head injury guidelines and rules has had a tremendous impact in decreasing head injury severity in football. Modifications of current testing models may further improve helmet design and hence further decrease the incidence and severity of head injury sustained while playing football. PMID- 15335434 TI - Hypoxia-inducible factor and vascular endothelial growth factor are expressed more frequently in embolized than in nonembolized cerebral arteriovenous malformations. AB - OBJECTIVE: In previous studies, we documented a marked neoangiogenesis and endothelial proliferation in cerebral arteriovenous malformations (AVMs) that were embolized before surgery compared with those that were not embolized. We hypothesized that embolization caused a local hypoxia that promotes neoangiogenesis as a possible pathomechanism. To support this hypothesis, we now examined the angiogenesis-related proteins in a larger cohort of patients. In addition, we investigated hypoxia-inducible factor-1 alpha as a possible protein operative during neoangiogenesis of cerebral AVMs. METHODS: Paraffin-embedded specimens of 56 AVMs obtained from surgical resection and 14 brain tissue controls were immunohistochemically stained with antibodies to proliferating cell nuclear antigen, MIB-1, vascular endothelial growth factor, Flk1, and hypoxia inducible factor-1 alpha by standard protocols. RESULTS: In AVMs treated with embolization before surgery (n = 35, 63%), the expression of hypoxia-inducible factor-1 alpha (P = 0.0101) and vascular endothelial growth factor (P = 0.0007) was significantly higher (Fisher's exact test) than in patients who did not have previous endovascular treatment. Differences in the expression of Flk-1 (P = 0.0798) and proliferating cell nuclear antigen (P = 0.0423) were in the same direction but were not significant when corrected for multiple testing. CONCLUSION: Our results provide circumstantial evidence that a partial occlusion of cerebral AVMs might induce local hypoxia-related neoangiogenesis. To support these data, future animal studies should be performed. PMID- 15335435 TI - Nitric oxide synthase in acute alteration of nitric oxide levels after subarachnoid hemorrhage. AB - OBJECTIVE: Subarachnoid hemorrhage (SAH) is associated with acute decreases and subsequent recovery of cerebral nitric oxide (NO) levels, but the mechanisms of these alterations are not known. In this study, we measured NO synthase (NOS) protein and kinetics to determine its involvement in the alterations of cerebral NO levels after SAH. METHODS: The endovascular rat model of SAH was used. The number of NOS-1 (neuronal) and NOS-2 (inducible)-positive cells (0-96 h) was determined by counting immunoreactive cells in 8-microm cryostat sections. The tissue content of active NOS and its kinetic parameters were studied with an enzymatic l-citrulline assay. RESULTS: The number of NOS-1-positive cells increased between 1 and 3 hours after SAH, decreased to and below control values at 6 and 72 hours after SAH, and increased to control values 96 hours after SAH. The number of NOS-2-positive cells increased 1 hour after SAH, decreased to control values at 24 hours, and increased above control values 96 hours after SAH. The Michaelis-Menten kinetic parameters (V(max), K(m), slope) of NOS remained unchanged at 10 and 90 minutes after SAH. CONCLUSION: NOS-1 and -2 proteins undergo a triphasic alteration after SAH, whereas the amount of active NOS and its kinetic parameters remain unchanged during the first 90 minutes after SAH. Depletion of NOS is not involved in the acute alterations of cerebral NO levels after SAH. PMID- 15335436 TI - Hypertonic fluid resuscitation from subarachnoid hemorrhage in rats. AB - OBJECTIVE: Increased intracranial pressure (ICP) and decreased cerebral blood flow leading to global cerebral ischemia are the primary causes of death after severe subarachnoid hemorrhage (SAH). Hypertonic saline has been demonstrated to exert neuroprotective properties after traumatic brain injury by osmotic mobilization of parenchymal water and improvement of microcirculation. We used a rat model to investigate the effects of hypertonic fluid resuscitation after SAH on ICP, cerebral blood flow, body weight, neurological recovery, and morphological damage. METHODS: Sixty rats were subjected to SAH induced by an endovascular filament. ICP and local cerebral blood flow were recorded continuously. Animals were assigned to three groups: 1) NaCl 0.9%; 2) NaCl 7.5% (4 ml/kg); and 3) NaCl 7.5% plus 6% dextran 70 (4 ml/kg) given 30 minutes after SAH. Body weight and neurological deficits were assessed daily. Morphological damage was evaluated on Day 7. RESULTS: SAH resulted in an immediate increase of ICP to approximately 60 mm Hg initially, and then to approximately 30 mm Hg for the next 90 minutes. Although NaCl 7.5% alone and in combination with dextran led to an immediate, significant, and lasting decrease of ICP to 15 to 20 mm Hg, only the combined therapy significantly increased body weight and improved neurological recovery. Furthermore, the group that received combined therapy exhibited significantly more surviving neurons in hippocampus, cortex, caudoputamen, and cerebellum. Mortality was reduced nonsignificantly, from approximately 65% in groups I and II to 35% in Group III. CONCLUSION: Treatment with NaCl 7.5% plus 6% dextran 70 is significantly effective for reducing the initial harmful sequelae of SAH. The regimen resulted in lowered ICP, improved neurological recovery, and less morphological damage after SAH in the rat. PMID- 15335437 TI - The history of neurosurgery at Temple University. AB - TEMPLE UNIVERSITY'S NEUROSURGERY program has had a colorful and distinguished history since its creation in 1929. It has always functioned under challenging circumstances with limited resources but with a strong sense of mission. It was one of the 20 neurosurgical training programs in existence when the American Board of Neurosurgery was founded in 1940. Over the past 64 years, the program has trained approximately the same number of neurosurgeons, many of whom have contributed significantly to our specialty. Some of the advances pioneered in part at Temple include clinical hypothermia (Fay), the biplanar stereoscopic angiographic unit (Chamberlain), human stereotactic surgery (Spiegel and Wycis), lumboperitoneal shunts (Scott), posterior lumbar interbody fusion (Lin), microsurgery for acoustic tumors (Buchheit), and new pharmacological approaches to neuroprotectors (Strauss and Narayan). The Temple neurosurgery program has survived many challenges in the past and will no doubt weather the current financial and medicolegal challenges that confront the neurosurgical community in Philadelphia. It remains a strong clinical program that serves an otherwise underserved community and attracts patients beyond its geographic area because of its strong clinical reputation and the excellence of its clinical faculty and residents. PMID- 15335438 TI - Surgery of involuntary movements, particularly stereotactic surgery: reminiscences. AB - WHEN WE STARTED using stereotactic surgery in 1953, the existing stereotactic instruments required general anesthesia for fixation of the head. We designed a stereotactic instrument with target screens that could be fixed to the patient's head under local anesthesia. The results of stereotactic surgery for spasmodic torticollis were inconsistent. The active muscles were identified by electromyography and blocked with 1% lidocaine, resulting in marked but temporary improvement. It was decided to use selective peripheral denervation in these cases with very satisfactory results, and it became the only procedure we used for the relief of spasmodic torticollis. PMID- 15335439 TI - Lumbar spine fracture in a 34,100-year-old skeleton: the oldest known prehistoric spine fracture. AB - WE EXAMINED SPECIAL pathological changes of the lumbar spine from skeleton remains referred to as "Stetten 1," one of the earliest specimens of modern humans dating from the Early Stone Age (Upper Paleolithic). The skeleton was discovered during archaeological excavations under Riek in 1931 in the Vogelherd Cave near Stetten, close to the Lone Valley of southwestern Germany. The archaeological context is the so-called Aurignacian period (40,000-30,000 yr ago), representing the earliest cultural step of modern humans in Europe. Accelerator mass spectrometry with (14)C measurement yields a calibrated age of these remains averaging 34,100 years before the present. The L3-L4 vertebrae exhibit marginal bone fusion on the right side with a smooth surface. They show 20- to 30-degree kyphosis secondary to wedge impaction of the L4 vertebral body. The facet joints and vertebral bodies reveal small marginal osteophytes with even joint surfaces, indicating low degenerative changes. Stetten 1 is characterized by the presence of a healed lumbar spine fracture. It documents the earliest known case thus far of spine fracture among modern humans. PMID- 15335440 TI - Frontotemporal dementia: genetics and genetic counseling dilemmas. AB - BACKGROUND: Frontotemporal dementia (FTD) is a neurodegenerative disease with early symptoms of personality change and/or language disorder. Approximately 40% of individuals with FTD have a family history of dementia; however, in our experience, less than 10% have clear autosomal dominant inheritance. Mutations in the microtubule-associated protein tau (MAPT) gene have been reported in up to 50% of hereditary cases, but are unusual except in families with more than 3 individuals with FTD. The genetics of FTD is complicated by clinical heterogeneity, variable expression, phenocopies, misdiagnoses, and lost family histories. The objective of this paper is to enable physicians to recognize hereditary patterns and genetic concerns of FTD families and to understand genetic counseling strategies. REVIEW SUMMARY: The complexity of FTD genetics and genetic counseling are illustrated using 4 case histories. Case 1 demonstrates the difficulty obtaining a reliable FTD family history. Case 2 illustrates how psychiatric phenocopies can make family linkage studies difficult. The lack of genotype and phenotype correlation and issues of predictive genetic testing within FTD families are the subject of case 3, and case 4 shows how normal aging language difficulties and cognitive changes can be misinterpreted when a family history of dementia is present. CONCLUSIONS: Physicians seeing patients with possible FTD should be aware of the risk of a genetic etiology. A 3-generation family history should be obtained with attention to neurologic, psychiatric, and behavioral symptoms. Variable expression and phenocopies are confounding factors when assessing a possible genetic etiology. Referral of the patient and family for genetic counseling is recommended. PMID- 15335441 TI - The motor cortex and facial expression: new insights from neuroscience. AB - BACKGROUND: For more than a century, unusual and complex deficits in facial expression have been known to occur following localized brain damage. Some brain injuries leave the face with pronounced alterations in affect whereas others result in movement disorders such as blepharospasm and Meige syndrome. There is also a historic trail of clinical observations that document deficits in either voluntary or emotional control of the facial muscles following central nervous system damage. REVIEW SUMMARY: Recent studies in the nonhuman primate cerebral cortex reveal the existence of multiple cortical facial representations in the frontal lobe and adjacent anterior cingulate cortex. These comprise the facial representation of the primary motor cortex (M1), ventral lateral premotor cortex (LPMCv), supplementary motor cortex (M2), rostral cingulate motor cortex (M3), and caudal cingulate motor cortex (M4). Homologous facial representations reside in the human brain based on observations following cortical stimulation, functional neuroimaging, and localized surgical resection. In the nonhuman primate, all these facial representations have been found to be directly interconnected through topographically organized corticocortical connections, and each facial area has also been found to send direct corticobulbar projections to the facial motor nucleus. The facial representations of M2 and M3 are both located on the medial wall of the hemisphere, in the vascular territory of the anterior cerebral artery. Both preferentially give rise to bilateral projections to parts of the facial nucleus that innervate the upper facial musculature as demonstrated in the monkey. The facial representation of M1, LPMCv, and M4 preferentially give rise to contralateral axonal projections ending in parts of the facial nucleus that innervate the lower facial musculature. The facial representation of M1 and LPMCv both reside in the vascular territory of the middle cerebral artery (MCA). The classic clinical presentation of paralysis in the contralateral lower facial musculature and intact bilateral upper facial musculature following typical MCA in infarction in the human parallels this mapping pattern of corticobulbar connections found in the nonhuman primate. CONCLUSIONS: Facial movements are undoubtedly under the powerful influence of the cerebral cortex and are essential for the appropriate execution of many important functions such as mastication, swallowing, and social interaction, including speech and nonverbal communication. This information provides a theoretic template for interpreting the clinical effects of neuropathologic disease and localized cortical trauma on facial movements. PMID- 15335442 TI - Essential tremor. Epidemiology, diagnosis, and treatment. AB - BACKGROUND: Essential tremor (ET) is the most common adult tremor disorder and is characterized by postural and kinetic tremor. Symptoms are typically progressive and potentially disabling, often forcing patients to change jobs or seek early retirement. Proper treatment is contingent on a correct diagnosis, and other possible causes of tremor must be excluded. REVIEW SUMMARY: Although primidone and propranolol have been regarded as the mainstays of pharmacologic therapy for ET, additional agents may be useful in reducing tremor. Surgical procedures are available that effectively ameliorate tremor that is refractory to medical management. This article reviews the epidemiology, pathophysiology, and treatment options for ET. CONCLUSIONS: Despite a range of treatment options currently available, further research is necessary to manage this syndrome most effectively. Double-blind, controlled trials are needed to determine whether primidone, propranolol, or a combination of these medications is superior in the initial management of ET. Other pharmacologic agents have shown potential to reduce tremor and should be investigated further. Additional studies are also needed to determine the best treatment of head and voice tremor with pharmacologic and surgical interventions. With proper treatment, tremor is sufficiently reduced in the majority of patients. PMID- 15335443 TI - Failed back syndrome. AB - BACKGROUND: Failed back syndrome (FBS) is a well-recognized complication of surgery of the lumbar spine. It can result in chronic pain and disability, often with disastrous emotional and financial consequences to the patient. Many patients have traditionally been classified as "spinal cripples" and are consigned to a life of long-term narcotic treatment with little chance of recovery. Recent progress in our understanding of this complex condition, as well as advances in surgical and medical treatment, have offered hope for improved outcomes and quality of life for these patients. REVIEW SUMMARY: This study explores the complex and multifactorial etiology of FBS, including reasons for surgical failure, indications for surgical revision, and adjunctive treatments such as pain management procedures. CONCLUSIONS: Despite extensive work in recent years, FBS remains a challenging and costly disorder. Recent advances in surgical reconstruction, rehabilitation, and pain management technique offer hope for patients with this painful and disabling condition. PMID- 15335444 TI - Anticipating smallpox and monkeypox outbreaks: complications of the smallpox vaccine. AB - BACKGROUND: The recent outbreak in the Midwest of monkeypox, as well as the continued fears of a terrorist-induced epidemic of smallpox, prompted the authors' review of the literature regarding past and current experiences with smallpox vaccination. The smallpox vaccine, which is highly effective in preventing the spread of both these orthopoxvirus infectious illnesses, might be administered to numerous health care workers and, in the event of a smallpox attack, millions of other citizens. However, vaccinees would be at risk for several vaccine-related neurologic complications. REVIEW SUMMARY: According to prior reports, neurologic complications have occurred in 2.5 per million US individuals, with the most common being postvaccinal encephalomyelitis (PVEM). In older children and adults, PVEM causes stupor and coma, seizures, paraparesis, and other neurologic and mental abnormalities, and, in 16% of cases, permanent neurologic sequelae. The overall mortality rate of neurologic complications is approximately 1.5 per million vaccinees. Risk factors for PVEM were age younger than 1 year and no previous smallpox vaccination, but not a prior episode of PVEM or other preexisting neurologic illnesses. Neither the current smallpox vaccination campaigns in Israel nor the one in the United States has had comparable complications, but the US campaign has been associated with myocarditis and myopericarditis. CONCLUSION: Although the potential neurologic complications of the smallpox vaccine must be weighed against the threat of monkeypox and smallpox, current experience with vaccination suggests it carries a very low risk of neurologic complications and does not lead to exacerbations of chronic neurologic illnesses. PMID- 15335445 TI - Symptomatic and disease-modifying therapy for the progressive ataxias. AB - BACKGROUND: The progressive ataxias are a diverse group of neurologic diseases that share features of degeneration of the cerebellum and its inflow/outflow pathways but differ in etiology, course, and associated noncerebellar system involvement. Some will have treatable causes, but for most, the pathophysiology is incompletely known. REVIEW SUMMARY: Treatment strategies will include (1) definitive therapy when available, (2) symptomatic treatment and prevention of complications, and (3) rehabilitation and support resources. The physician will have to decide whether to introduce or approve the use of therapies based on as yet-unproven mechanisms or the use of complementary medicine approaches. CONCLUSIONS: There are as yet no drugs that have been approved by the Food and Drug Administration for the treatment of the progressive ataxias and relatively few disease-modifying therapies, but symptomatic and rehabilitation interventions can greatly improve the quality of life of individuals with these disabling neurodegenerative disorders. PMID- 15335446 TI - A mnemonic for Parkinson disease patients considering DBS: a tool to improve perceived outcome of surgery. AB - Patients considering deep brain stimulation (DBS) for Parkinson disease (PD) may be exposed to videotapes, media coverage, or literature which show dramatic improvements in PD symptoms after surgical intervention. Based on this information, patients may seek a medical center with expertise in DBS for an evaluation and assessment of their candidacy for surgery. If patients receive a device, they may be disappointed or despondent following surgery because of a failure to achieve a preconceived and unrealistic outcome. In order to address the important issue of patient misconception of potential outcome, we have introduced a simple mnemonic device. The device may be taught and then reviewed with patients and families both before and after surgery. Use of this mnemonic device may allow the patient and family the time necessary to alter the perception of perceived benefit. This education can help to ensure that outcome meets or exceeds expectation, and as a result they become a more satisfied and easy-to-manage DBS patient. PMID- 15335447 TI - On not being the "hostess with the mostest". PMID- 15335450 TI - Adjuvant therapy in elderly patients with breast cancer. AB - The elderly population has been neglected by the traditional approach to clinical breast cancer research. Elderly women have been underrepresented in breast cancer clinical trials, with the majority of studies being restricted to patients aged < 70 years. Elderly patients frequently have comorbidities and/or impaired organ function. These facts may often lead to death from causes other than cancer, thus nullifying any possible benefit of adjuvant treatment; furthermore, they render extrapolation of standard treatment recommendations to the elderly potentially hazardous, particularly with respect to chemotherapy. Therefore, specific clinical trials are needed to investigate adjuvant treatments tailored for the heterogeneous older population. PMID- 15335452 TI - Preoperative chemotherapy and endocrine therapy in patients with breast cancer. AB - This review focuses on the aims, results, advantages, and possible disadvantages of preoperative chemotherapy and endocrine therapy. We present the recent improvements in terms of pathologic response rates that have resulted from new combinations of drugs. The change of established prognostic factors during neoadjuvant treatment, the need for new markers, and the consequences in terms of clinical decision-making are demonstrated. We discuss the risk of local relapse after breast-conserving surgery, which was made feasible by preoperative chemotherapy. A short overview of current neoadjuvant cytostatic, endocrine, and immunotherapy trials is provided. Future opportunities for tailoring therapy to each individual patient based on early information from the primary tumor are discussed. Important considerations and results of recent endocrine trials that analyzed possible tamoxifen-resistance in subgroups are reported. New opportunities exist to evaluate the efficacy of new cancer drugs more rapidly in the neoadjuvant setting than in the metastatic and adjuvant setting. This approach offers the possibility of monitoring prognostic markers in the primary tumor before, during, and after treatment with specific chemotherapeutic agents. With respect to recent findings of gene-array techniques, it is likely that the advances in this technology will lead to improved prognostic statements. It will show the influence of therapy on gene expression profiles in the course of treatment and might enable us to identify chemoresistance of specific tumors rather early. This could potentially lead to a new direction of cancer therapy. PMID- 15335453 TI - Phase II trial of a doxorubicin/docetaxel doublet for locally advanced and metastatic breast cancer: results from national surgical adjuvant breast and bowel project trial BP-57. AB - A phase II trial at 12 institutions using AT (doxorubicin 60 mg/m2 plus docetaxel 60 mg/m2) given every 21 days was conducted. Eighty-nine patients were entered who ranged in age from 25 to 75 years, 41.6% of whom had stage IIIB disease and 58.4% of whom had stage IV disease. Among the patients with stage IV disease, 32.7% had received prior adjuvant chemotherapy. Premedication with dexamethasone (8 mg orally twice per day for 3 days) and prophylactic ciprofloxacin (500 mg orally twice per day on days 5-15) was used. Colony-stimulating growth factors were reserved for secondary prophylaxis after prolonged or febrile neutropenia (FN) or documented severe infection in an earlier cycle. After a cumulative dose of doxorubicin of 480 mg/m2, patients could continue to receive docetaxel (100 mg/m2) alone. Median time on study as of July 6, 2003, was 54 months. Febrile neutropenia occurred in 36 patients (41.9%): 23 developed FN in the absence of previous prophylactic growth factor support and 13 developed it despite previous growth factor support. One patient died from sepsis. Other grade 3/4 adverse events included nausea in 3.5%, vomiting in 4.7%, stomatitis in 8.1%, diarrhea in 5.8%, arthralgia/myalgia in 2.3%, fluid retention in 1.2%, pulmonary embolism in 1.2%, rest dyspnea in 1.2%, neuromotory toxicity in 1.2%, and neurosensory toxicity in 1.2%. Clinical congestive heart failure was seen in 2 patients (2.3%). Sixty-seven patients were evaluable for best response with 6 cycles of therapy. Fourteen patients (20.9%) had a complete response and 30 (44.8%) had a partial response, for an overall response rate of 65.7% in evaluable patients. The median response duration was 25.9 months, and the median time from entry to progression or death was 27.5 months. The median survival time for the 86 patients with endpoint information was 31.1 months. The administration of AT with primary ciprofloxacin and secondary colony-stimulating factor prophylaxis is feasible, and the combination is active. Its value in the adjuvant setting is currently under investigation. PMID- 15335454 TI - Quality of life in women with advanced breast cancer treated with docetaxel. AB - A survey was conducted to address the question of whether overall quality of life (QOL) in women with advanced breast cancer is significantly impaired by treatment with docetaxel in routine clinical practice. Health-related QOL (HRQOL) measured by the European Organisation for Research and Treatment of Cancer QOL Questionnaire C30, as well as performance status (PS) and tumor response, were assessed at baseline, then after every 2 cycles of treatment. The 209 evaluable patients received a median of 5 treatment cycles (range, 2-9), with a response rate of 35% (95% CI, 28.5%-41.4%). There were significant adverse changes in fatigue (P = 0.002) and diarrhea (P = 0.0002). However, HRQOL scores for emotional function (P < 0.0001), pain (P = 0.0001), constipation (P = 0.01), and nausea and vomiting (P = 0.03) all showed significant beneficial changes. Several QOL parameters improved during docetaxel therapy even in patients who did not have an objective tumor response. Overall, PS was maintained during docetaxel treatment. Following the encouraging QOL data from clinical trials of docetaxel in breast cancer, the authors conclude that, in actual clinical practice, docetaxel does not have an overall detrimental effect on QOL. PMID- 15335456 TI - Parity confers better prognosis in older women with early-stage breast cancer treated with breast-conserving therapy. AB - A comparison was made of pretreatment characteristics and outcomes of patients with stage I/II breast cancer treated with breast-conserving therapy who had a history of parity with those who were nulliparous. From 1979 to 1996, 1358 women with stage I/II (T1/2 N0/1 M0) breast cancer underwent lumpectomy, axillary dissection, and radiation therapy with or without systemic therapy. Of the total population, 1162 patients (86%) were parous and 196 patients (14%) were nulliparous. The median follow-up was 87 months. The 2 groups were compared for clinical, pathologic, and treatment-related factors. Multivariate analysis was used to determine independent predictors of outcome. Outcome was also evaluated for patterns of failure including distant metastases (DM), cause-specific survival (CSS), and overall survival (OS). Significant differences between the 2 groups were observed for age > 60 years and median age. Multivariate analysis demonstrated that nulliparous status was an independent predictor of DM, CSS, and OS after adjusting for age. Multivariate analysis for DM, CSS, and OS for patients > 60 years of age demonstrated that parity was the most highly significant independent predictor of decreased DM and improved OS. Parity can be considered a prognostic factor in elderly patients with early-stage breast cancer, and therefore may be used as a tool for identifying patients who may benefit from a more aggressive treatment approach. PMID- 15335457 TI - HER2 and central nervous system metastasis in patients with breast cancer. AB - We report a case of a patient who developed a central nervous system (CNS) metastasis 17 months after surgery and adjuvant therapy for stage II breast cancer. The HER2 status of the primary tumor as determined by immunohistochemistry (IHC) was 1+, and fluorescence in situ hybridization (FISH) performed retrospectively showed a borderline-negative result (ratio, 1:9), although HER2 amplification was present in a minority of breast cancer cells. By contrast, the HER2 status of the CNS metastasis was positive by IHC (3+) and FISH (ratio, 3:7). This report discusses the possibility that, for metastatic invasion into the CNS, clonal selection may occur, favoring HER2-positive cell clones. PMID- 15335458 TI - Pathologic changes in breast cancer following neoadjuvant chemotherapy: implications for the assessment of response. AB - Neoadjuvant chemotherapy (also known as preoperative or primary chemotherapy) is the treatment of choice for patients with locally advanced breast cancer. One of the main advantages of neoadjuvant chemotherapy is that it allows for assessment of pathologic response to treatment. Clinical and radiologic evaluations of response to neoadjuvant chemotherapy are based on change in tumor size, and the correlation with pathologic response is often inaccurate. Pathologic evaluation of tumor size remains the gold standard for evaluation of residual tumor after chemotherapy. Chemotherapy-induced histomorphologic change is commonly observed in posttreatment resection specimens and can contribute to the less-than-perfect correlation between the clinical assessment of tumor size and the pathologic measurement. Therefore, accurate histologic mapping to the macroscopic and radiologic appearance of the tumor bed is necessary. Cytopathologic changes are also common in residual cancer cells after neoadjuvant chemotherapy and have uncertain clinical relevance. There is a role for the development of new histologic approaches to augment the pathologic and clinical assessment and provide information on the differential response, particularly for tumors in which less than pathologic complete response is achieved. PMID- 15335459 TI - Why learning and development can lead to poorer recognition memory. AB - Current models of inductive reasoning in children and adults assume a central role for categorical knowledge. A recent paper by Sloutsky and Fisher challenges this assumption, showing that children are more likely than adults to rely on perceptual similarity as a basis for induction, and introduces a more direct method for examining the representations activated during induction. This method has the potential to constrain models of induction in novel ways, although there are still important challenges. PMID- 15335460 TI - Understanding culture across species. AB - Recent claims of culture in great apes have provoked fervent argument about the 'true' definition of culture, most of which has been unhelpful. Instead, a range of definitions should be used to explore different aspects of the cognitive processes that together result in human culture, many of which can be productively studied in non-humans. A richer cognitive account of the contents of culture needs to be developed and used to compare animal and human cultures, instead of sterile searching for a cognitive Rubicon between them. Exploring six views of culture, this article highlights the fundamental contrast of whether culture evolves as a by-product of cumulative change in cognitive mechanisms, or whether it is actively selected for its advantages. PMID- 15335461 TI - Cognitive functions of gamma-band activity: memory match and utilization. AB - Oscillatory neural activity in the gamma frequency range (>30Hz) has been shown to accompany a wide variety of cognitive processes. So far, there has been limited success in assigning a unitary basic function to these oscillations, and critics have raised the argument that they could just be an epiphenomenon of neural processing. We propose a new framework that relates gamma oscillations observed in human, as well as in animal, experiments to two underlying processes: the comparison of memory contents with stimulus-related information and the utilization of signals derived from this comparison. This model attempts to explain early gamma-band responses in terms of the match between bottom-up and top-down information. Furthermore, it assumes that late gamma-band activity reflects the readout and utilization of the information resulting from this match. PMID- 15335462 TI - Vitalistic causality in young children's naive biology. AB - One of the key issues in conceptual development research concerns what kinds of causal devices young children use to understand the biological world. We review evidence that children predict and interpret biological phenomena, especially human bodily processes, on the basis of 'vitalistic causality'. That is, they assume that vital power or life force taken from food and water makes humans active, prevents them from being taken ill, and enables them to grow. These relationships are also extended readily to other animals and even to plants. Recent experimental results show that a majority of preschoolers tend to choose vitalistic explanations as most plausible. Vitalism, together with other forms of intermediate causality, constitute unique causal devices for naive biology as a core domain of thought. PMID- 15335463 TI - How parallel is visual processing in the ventral pathway? AB - Visual object perception is usually studied by presenting one object at a time at the fovea. However, the world around us is composed of multiple objects. The way our visual system deals with this complexity has remained controversial in the literature. Some models claim that the ventral pathway, a set of visual cortical areas responsible for object recognition, can process only one or very few objects at a time without ambiguity. Other models argue in favor of a massively parallel processing of objects in a scene. Recent experiments in monkeys have provided important data about this issue. The ventral pathway seems to be able to perform complex analyses on several objects simultaneously, but only during a short time period. Subsequently only one or very few objects are explicitly selected and consciously perceived. Here, we survey the implications of these new findings for our understanding of object processing. PMID- 15335464 TI - Mechanisms of theory formation in young children. AB - Research suggests that by the age of five, children have extensive causal knowledge, in the form of intuitive theories. The crucial question for developmental cognitive science is how young children are able to learn causal structure from evidence. Recently, researchers in computer science and statistics have developed representations (causal Bayes nets) and learning algorithms to infer causal structure from evidence. Here we explore evidence suggesting that infants and children have the prerequisites for making causal inferences consistent with causal Bayes net learning algorithms. Specifically, we look at infants and children's ability to learn from evidence in the form of conditional probabilities, interventions and combinations of the two. PMID- 15335465 TI - Computational approaches to the development of perceptual expertise. AB - Dog experts, ornithologists, radiologists and other specialists are noted for their remarkable abilities at categorizing, identifying and recognizing objects within their domain of expertise. A complete understanding of the development of perceptual expertise requires a combination of thorough empirical research and carefully articulated computational theories that formalize specific hypotheses about the acquisition of expertise. A comprehensive computational theory of the development of perceptual expertise remains elusive, but we can look to existing computational models from the object-recognition, perceptual-categorization, automaticity and related literatures for possible starting points. Arguably, hypotheses about the development of perceptual expertise should first be explored within the context of existing computational models of visual object understanding before considering the creation of highly modularized adaptations for particular domains of perceptual expertise. PMID- 15335466 TI - Fitness for interview: current trends, views and an approach to the assessment procedure. AB - The medical assessment of a person's fitness to be interviewed by police is undertaken to determine the detainee's competence at interview. A review of this procedure in Victoria was conducted on all cases during the period 1 January 30 June 1997. One hundred and fifty-one cases were assessed. The majority of cases were in the 20-39 years age range with 81.5% of all cases being male. The principal reasons for assessment were concerns about the effects of drugs excluding alcohol (21.9%), and psychiatric issues (25.8%). Following assessment, 47% were considered to be fit for interview but in 14% of these cases an independent third person was recommended. Thirty-five per cent were assessed as being unfit for interview and 58.6% of these were referred for further assessment. Rarely (4.6%) in these cases and those referred for treatment elsewhere were reviews arranged prior to the interview. Despite the significant dependence of the prosecution on confessional evidence, in only 4% of cases was the matter eventually raised in court. The findings indicate that there is a need to ensure that practitioners are appropriately trained in the assessment of drug affected and psychiatric patients. New guidelines for the assessment are presented which may assist in reducing the degree of subjectivity and opinion variability. PMID- 15335467 TI - Cost of drug use and criminal involvement before and during methadone treatment. AB - AIMS: To examine criminal behaviour in methadone clients as a function of expenditure on illicit drugs. To quantify the benefit of treatment for this group and to identify individuals who do not reduce criminal behaviour or substance use as a consequence of treatment. DESIGN: Cross-sectional examination of key methadone-treatment variables (age, methadone dose and length of time in treatment), and self-reported levels of spending on illicit drugs before and during methadone treatment. SETTING: All interviews were conducted in the community-based treatment centre of the Maudsley's addiction treatment services in South London. PARTICIPANTS: One hundred consecutive opiate-dependent attenders at a community treatment clinic. MEASUREMENTS: Data were collected using a self report questionnaire and treatment characteristic data were collected from clinical case notes. FINDINGS: There was a major overall reduction in spending on illicit drugs from before treatment to during treatment, consistent with previous findings; however, 15% of the sample either showed no change or actually increased their spending after entry to treatment. This group did not differ in terms of key treatment characteristics (age, methadone dose and length of time in treatment) and there was no significant correlation between levels of spending before and during treatment. CONCLUSIONS: Despite substantial overall benefit, there is a subgroup of individuals for whom long-term treatment, even on high range doses of methadone, has no beneficial effect on criminal activity or drug use. Further research is required to establish the dynamics of this indirect treatment-benefit, and to explore the apparent resistance to treatment of the criminal behaviour of a small sub-group. PMID- 15335468 TI - Acute myocardial ischaemia presenting as sudden death in the Lothian and Borders regions of Scotland, 1987 and 1997. AB - OBJECTIVES: To profile deaths from acute myocardial ischaemia presenting as sudden unexpected death in the years 1987 and 1997, and to review the diagnosis of those patients who sought medical attention. DESIGN: A retrospective study using data obtained from autopsy and police reports held in the Forensic Medicine Unit of the University of Edinburgh. RESULTS: Cardiovascular pathology was present in a significant proportion of cases. Clinically 'silent' events presenting as a sudden, unexpected death were common, comprising more than one half of the events observed in 1987 and one-third of those in 1997. For symptomatic cases, respiratory and upper gastrointestinal symptoms were the most common presentation. The majority of patients seeking medical advice were misdiagnosed as either having a chest infection or indigestion. CONCLUSIONS: Doctors in the primary care setting should include acute myocardial ischaemia as a possible differential diagnosis of a number of symptoms which initially do not suggest a myocardial origin. PMID- 15335469 TI - Incidence trends in house fire fatalities in eastern Scotland. AB - Fatal house fires occur more frequently in Scotland than elsewhere in Great Britain.(1,2) Over the last few years several educational campaigns were initiated to instruct the public in the risk factors associated with such fire fatalities and to promote the installation of functional domestic smoke detectors. An epidemiological survey of house fire deaths in the Lothian and Borders region of Scotland over the first half of the present decade, was carried out to evaluate any changing incidence trends by comparison with a survey of the fatalities from the first half of the previous decade. The impact of such campaigns in reducing house fires appears to be limited. PMID- 15335470 TI - Investigation of clandestine graves resulting from human rights abuses. AB - The role of the forensic scientist in the investigation of clandestine graves resulting from human rights abuses is discussed. Individuals buried in such circumstances have perished following abuse by persons in authority or by other political factions. Such persons have been classed by the abusers as dissidents or political opponents, or have been killed simply for belonging to a particular racial or religious grouping. Guidelines for investigation and the general approach by the scientist are suggested and examples of exhumations are cited in which the author has had personal involvement. These include exhumations carried out in the former Yugoslavia, Rwanda and Chile. PMID- 15335471 TI - Delayed rupture of spleen in a case of spouse abuse. AB - This case deals with assault of a housewife by her husband. Although there had been previous episodes of abuse, the gravity of the present injury, which required hospitalization, resulted in the fact being made known to others. Insistence on a wife's right to protection and shelter from a husband who assaults her has grown significantly in the wake of the women's liberation movement. However, it remains the case that few women in Sri Lanka would take action and risk disrupting their families. PMID- 15335472 TI - Male genital mutilation: 'whodunnit'? AB - A 26-year-old man was brought to hospital having sustained amputation of his penis, scrotum and testes. Although he claimed that his 'master' did it, the 'master' denied the allegation. The police subsequently retrieved the two testes but the amputated penile shaft was never found. The remaining scrotum and short penile stump were repaired to enable him pass urine. The evidence of the patient was not corroborated. No motive for the amputation was established. The patient refused to appear in court as prosecution witness against the accused, his 'master'. Furthermore, although he has not withdrawn his accusation, he has paid friendly visits to the master subsequently. There is, therefore, the suspicion that the patient was himself responsible for the amputation. This patient will require long-term psychological, endocrinological and urological follow-up. PMID- 15335473 TI - Human rights and the rule of law. AB - This personal view was first delivered to the 5th Conference of the World Police Medical Officers in Clinical Forensic Medicine, in Vancouver, Canada, in August 1999 by the Right Honourable Sir Robert Carswell, Lord Chief Justice of Northern Ireland. PMID- 15335474 TI - Forensic web watch. AB - Now that one has logged onto the world wide web (WWW) and utilized one or more of the home pages listed previously (or used another equally good home page) to seek out basic information available to forensic practitioners, the question now arises of how to go about making the most of the information available. One feature consistent to most home pages is links to the home pages of Associations and Societies, one or more of which most practitioners will be members of. With access to the WWW not only have you access to your own association/society, but you can also keep up to date with all the others to which you have not paid subscriptions. Although an internet search using a WWW search engine or the 'top 6' home pages may assist in identifying a large number of association and society sites, one of the most useful places to start is the home page of the Indian Academy of Forensic Medicine (IAFM). This, to date, lists a total of 139 such sites. To access all the home pages listed may take in excess of 6 h so the following review looks at the range of sites available and recommends some places the author considers many people may wish to know and visit. Again, this is inevitably a personal choice and it is recognized that those sites not listed may, in fact, be the preferred choice for other users of the forensic WWW. PMID- 15335475 TI - Massive project. PMID- 15335476 TI - Examination for the late physical after effects of torture. PMID- 15335479 TI - Continuing Education Programme - Victorian Institute of Forensic Medicine and Department of Forensic Medicine, Monash University. AB - Given the increasing requirement of the courts for forensic experts to engage in ongoing education, a continuing education programme was developed in forensic medicine at the Victorian Institute of Forensic Medicine to cater for both clinicians and pathologists. The programme consists of a series of cases which are circulated several times per year. All are actual cases and are reflective of the types of presentations experienced in forensic medicine. Each case includes relevant and appropriate details/findings and may include photographs. A series of questions follow which are answered usually in short-answer format. The answers are returned and correlated by a review panel, and a commentary with the response outcome is distributed to all those involved. The cases presented here are selected from this programme. PMID- 15335480 TI - Occupational fatalities in Israel. AB - Occupational fatal injuries are a worldwide problem. Certain occupations pose a greater risk than others. The purpose of this study was to estimate the death rates from occupational injuries in Israel over a period of 30 months, and to examine the trends in the light of the large numbers of foreign workers who have been brought into the country in the last decade. Two-thirds of the occupational fatalities were in the construction business, mostly owing to falls from a height resulting in death from multiple trauma. About one-third of these victims were foreign workers, even though they comprise only 20% of the work force in this field. Negligible amounts of alcohol were detected in a number of cases. The construction industry is recognized worlwide as a high-risk area claiming many lives each year. The findings in this study suggest that too little attention is given to safety regulations in general, and particularly concerning foreign workers in Israel. PMID- 15335481 TI - Drugs and driving: a retrospective study of the analyses of blood and urine specimens submitted to the Lothian and Borders Police Forensic Laboratory. AB - A comprehensive review of 75 samples of both blood and urine from 72 different drivers suspected of being impaired by drugs in the Lothian and Borders Police Force, Scotland area of jurisdiction, between 1st January 1995 and 2nd May 1997, was undertaken. The police reports, analytical results and criminal conviction records relating to each of the drivers were examined. This provided useful information concerning differences in laboratory procedures and produced a profile of the average drugged driver. The average age of the drivers was 23 years. Only two females were within the sample. Drugs were found in 65 cases (86.7%). Polydrug use was found in seven cases (9.3%). The drugs found, in order of frequency, were benzodiazepines (40%), cannabinoids (24%), alcohol (16%), methadone (12%), dihydrocodeine (9.3%), ecstacy (5.3%), amphetamine (2.7%), volatiles (1.3%) and morphine (1.3%). 90.3% of the drivers had previous convictions for criminal offences and 47.2% had convictions for drugs-related offences. Recommendations concerning police and medical training are discussed with particular reference to the Drug Recognition Expert program. PMID- 15335482 TI - Reliability of clinical decisions regarding alcohol influence. AB - In Jordan, pursuing legal procedures related to alcohol intake usually depends on the physicians' clinical judgment. A study was carried out on 825 medico-legal cases over a 2 year period to evaluate the extent of physicians' clinical decisions on patients suspected to be under the influence of alcohol. A significant number of cases (12.6%, n = 104) were found to be positive for alcohol. The study showed that the clinical examination failed to diagnose a large number of cases (78.9%, n = 82). Also, the majority of diagnosed cases (50%, n = 11) had a blood alcohol concentration (BAC) range of 50-100 mg/dl. 32% (n = 33) of the cases with positive BAC were admitted to hospital. A high admission rate was observed when BAC was <50 mg/dl and >150 mg/dl, which constituted 39.3% (n = 13) and 48.5% (n = 16) of the admitted cases respectively. A large number of the admitted cases had either simple or no trauma (48.4%, n = 16) as compared with the control group of cases (21.8%, n = 10). The admission rate of positive BAC cases in traffic accidents was double that of control group. These findings may reflect the unreliability of the clinical decision in positive BAC. Implementing a certain BAC level locally for both clinical and a jurisdiction purposes is strongly recommended. PMID- 15335483 TI - Clinical findings in the medico-legal investigation of arsonists. AB - Criminal and pathological arson is often committed with the help of fire accelerants. The formation of an explosive gas/air mixture may lead to an explosion whose extent and fierceness is often underestimated by the offenders. This explosion causes typical heat changes on exposed persons: * Singeing of eyelashes, eyebrows and hair growing near the frontal hairline * Singeing of the beard * Singeing of body hair in regions not covered by clothing (e.g. back of the hand) * In severe cases, skin burns of varying degree (back of hand, face). Such heat-related changes can sometimes be detected weeks after the offence. PMID- 15335484 TI - Genital warts in children: problems of management. AB - Anogential warts are much less common in children than in adults. In children the presence of anogenital warts have been suggested as an indicator for sexual abuse because of the association of genital warts with sexual activity. Difficulties of establishing mode of transmission arise because of the long latency of human papillomavirus and possibility of vertical and non-sexual transmission. A multidisciplinary approach is advised for the proper care of children with anogenital warts. PMID- 15335485 TI - Contact gunshot wound of the head: diagnosis after surgical debridement of the wound. AB - A case of homicidal contact gunshot wound of the head is described. The victim survived approximately 1 day and one-half following the shooting, during which time the bullet was recovered during surgery, the injuries were 'well documented', and surgical debridement of the wound destroyed any visible evidence of gunshot residue on the scalp or underlying bone. Autopsy revealed a skull fragment with soot deposition within the depths of the injured brain, allowing an accurate determination of range-of-fire. The case reiterates the importance of performing autopsies on all homicides. PMID- 15335486 TI - Toxic confusional state presenting as mental illness. AB - A 44-year-old man was arrested under section 136 Mental Health Act 1983 after behaving strangely outside a club. When assessed he appeared to be suffering from a toxic confusional state. On admission to hospital his salicylate levels were found to be high. The confusion proved to be caused by an aspirin overdose. PMID- 15335487 TI - Audit Commission: The Doctor's Bill Seminars - June/July 1998. PMID- 15335488 TI - Forensic web watch. AB - Since the development of the Internet (Net) by the Americans in the 1970s as a potential means of communication following a future world war, it has evolved over the decades into the graphical format of the world-wide web (WWW, Web) that we know today. Anyone accessing the Net, be it from home or work, has access to information and resources on almost anything but with this comes the time consuming and potentially expensive task of identifying those sites of use in one's professional work from those of purely interest or fun. These reviews will try and direct those working within forensic practice in all professions towards sites considered by the author to be practical resources. They are, by their very nature, a personal opinion and the author recognizes that the review of a site by one person may differ considerably from that of another user or the site designer. The articles will assume a basic knowledge of how to access and use the WWW using either Netscape or Microsoft Explorer. They will address groups of sites related to common areas of practice listing key web addresses to allow those interested to access sites reviewed. Because of the time between writing and publishing some sites may no longer be valid PMID- 15335489 TI - Driving whilst unfit through drugs. PMID- 15335491 TI - Re: The rate of elimination of alcohol. PMID- 15335490 TI - 'Criminalising' medical conditions. PMID- 15335492 TI - Continuing education programme - Victorian Institute of Forensic Medicine and Department of Forensic Medicine, Monash University. AB - Given the increasing requirement of the courts for forensic experts to engage in ongoing education, a continuing education programme was developed in forensic medicine at the Victorian Institute of Forensic Medicine to cater for both clinicians and pathologists. The programme consists of a series of cases which are circulated several times per year. All are actual cases and are reflective of the types of presentations experienced in forensic medicine. Each case includes relevant and appropriate details/findings and may include photographs. A series of questions follow which are answered usually in short-answer format. The answers are returned and correlated by a review panel, and a commentary with the response outcome is distributed to all those involved. The cases presented here are selected from this programme. PMID- 15335493 TI - An ophthalmological view of retinal haemorrhages in shaken babies. PMID- 15335494 TI - Under-reporting of suicide in South Yorkshire (West): a retrospective study of suicide and open verdicts returned by HM Coroner, 1992-1997. AB - A retrospective study was performed using information obtained from HM Coroner for South Yorkshire (West) over the period 1992-1997. All inquests which received a suicide or open verdict were reviewed to investigate the regional suicide statistics and the actual number of cases. Particular attention was paid to the age, sex, mental health and mode of death. There were 295 deaths believed to be suicidal in nature. Of these only 79% had suicide verdicts given and these comprise the official statistics. Of the open verdicts, many were suggestive of suicide but the circumstances were insufficient for the Coroner to be satisfied beyond reasonable doubt of the victims intention to kill themselves. Males and females had the same proportion of verdicts (80 and 76% respectively). Hanging was the commonest mode of suicide, (sex ratio 4.06 male: 1 female) and received a high percentage of suicide verdicts (81.5%). Self-poisoning was second but received suicide verdicts in only 66.3% of cases, whilst carbon-monoxide poisoning had a 98.1% rate of suicide verdict. Of the 295 cases, 202 decedents (88.5%) had a history of mental illness. Depression accounted for 153 cases (75.7%). Twelve decedents had psychotic illness (4%), one-half dying by jumping from a height. Official national data may under-report the annual suicide rate by over 20%. Deaths from poisoning by solid or liquid and jumping from a height appear particularly difficult to establish as suicide in coronial practice, with one-third of cases declared an open verdict. PMID- 15335495 TI - An analysis and report of custodial deaths in Nebraska, USA. AB - All medico-legal cases from the Lancaster County Coroner's Office in Lincoln, Nebraska USA, between 1991 and 1996, were reviewed for deaths which occurred in custody. The leading causes of death, in ranked order include: natural 45.1% (23/51), suicide 33.3% (17/51), homicide 11.8% (6/51), accident 7.8% (4/51), and undetermined 2% (1/51). Each cause of death was explored using the following variables: cause of death, age, sex, and race of the decedents. The findings of this investigation are compared with previous studies in other geographic regions of the country. Many similarities and differences in the frequencies of the modes of death are discussed. In addition, six unusual case studies including one accidental death, one homicide, and four suicides are presented and may serve as a reference for pathologists, forensic physicians and other forensic scientists confronted with aberrant cases. PMID- 15335496 TI - Age determination by teeth examination: a comparison between different morphologic and quantitative analyses. AB - Age determination by teeth examination is one of the main means of determining personal identification. Current studies have suggested different techniques for determining the age of a subject by means of the analysis of microscopic and macroscopic structural modifications of the tooth with ageing. The histological approach is useful among the various methodologies utilized for this purpose. It is still unclear as to what is the best technique, as almost all the authors suggest the use of the approach they themselves have tested. In the present study, age determination by means of microscopic techniques has been based on the quantitative analysis of three parameters, all well recognized in specialized literature: 1. dentinal tubules density/sclerosis 2. tooth translucency 3. analysis of the cementum thickness. After a description of the three methodologies (with automatic image processing of the dentinal sclerosis utilizing an appropriate computer program developed by the authors) the results obtained on cases using the three different approaches are presented, and the merits and failings of each technique are identified with the intention of identifying the one offering the least degree of error in age determination. PMID- 15335497 TI - Rape homicide involving children. AB - Three rape homicide cases and two cases of attempted rape homicide involving children investigated in the Bonn University Institute of Forensic Medicine from 1992 to 1996 are described in order to assess whether characteristic features were present. The victims were girls aged from 3 to 12 years old. At autopsy haematomas, contusions and/or lacerations of the external/internal genitalia or anus confirmed sexual assault. Extragenital injuries found in every case included bruises, haematomas, abrasions and incised wounds caused by resisting, to struggling and manual pressure or gripping. Each death was caused by manual or ligature strangulation. The evidential value of the anogenital injuries encountered at autopsy is discussed and compared with the physical signs found at clinical examination. PMID- 15335498 TI - Surfing the web or drowning in data: a subjective analysis of site and information availability on forensic-related subjects on the internet. AB - A subjective analysis of information available on the Internet to those working in forensic medicine medicine was performed. Internet web sites addressing forensic pathology, forensic science, anthropology, entomology, law enforcement, societies and journals were assessed. Ease of access, presentation, structure, specialization, links and popularity were reviewed. Each individual site assessed was graded out of 100 and ranked, by subject, to provide the reader with a generalized guide to what the authors considered was useful and/or interesting forensic sites on the internet. The need for a regulatory body for forensic Internet resources is discussed. PMID- 15335499 TI - Elevated blood ethanol levels caused by 'non-alcoholic' beer. AB - With the availability of non-alcoholic beer, many patients are substituting non alcoholic beer for regular beer. Moreover, several clinicians advocate the use of non-alcoholic beer by alcoholics when there is a possibility of relapse. However, patients with alcoholic end-stage liver disease can accumulate substantial circulating ethanol levels even when they consume non-alcoholic beer. We present a case of a patient with end-stage liver disease who achieved a blood alcohol level of 57 mg/dL after drinking non-alcoholic beer. PMID- 15335500 TI - Morphine overdose in a newborn and the search for the third order 'why?'. AB - A premature infant with pneumothorax-complicating respiratory distress syndrome died following a massive (100 fold) morphine overdose prior to insertion of an intercostal drain. The subsequent inquest considered the circumstances in which this error occurred and an open verdict was recorded. Adverse drug events remain an important cause of morbidity and mortality in hospital. A systems analysis approach to such incidents has been advocated as the most likely way to help minimize the risk of drug errors. PMID- 15335501 TI - Continuing education programme - Victorian Institute of Forensic Medicine and Department of Forensic Medicine, Monash University. PMID- 15335502 TI - Drug administration in custody. AB - This paper describes the system for drug administration to prisoners in custody which has been developed at U Division, Kilmarnock in Strathelyde. The protocols agreed by the police surgeons in the area are described. The system is relevant to all police surgeons in their daily work, particularly with the advent of general practice cooperatives and the decline in personal carriage of all but the most acutely required drugs. PMID- 15335503 TI - Examination for the late physical after effects of torture. AB - This paper describes the experience of doctors at the Medical Foundation for the Care of Victims of Torture, London, in documenting torture as recounted by survivors, mostly refugees from one of 63 countries, and in writing medico-legal reports to support their asylum claims. More than 2000 new patients are seen each year and 500 reports written. Few patients are seen in the acute phase; many not until 10 years following their torture. Patterns of torture practised in different countries are described, with the resulting physical signs. Methods of history-taking and examination are discussed, with emphasis on the detection of subtle forms of torture, recognition of deception and encouragement of full disclosure. PMID- 15335504 TI - Dental maturity of children in Perth, Western Australia, and its application in forensic age estimation. AB - The dental maturity of 1450 healthy children from Perth, Western Australia, was determined in a cross-sectional study. Orthopantomographs of 690 males and 760 females aged 4-16 years old, were randomly selected from various dental practices and hospitals in the Perth metropolitan area. Dental maturity was determined by the method of Demirjian and Goldstein based on the development of four permanent mandibular teeth. Curves were constructed for the Perth children and then compared to that of Demirjian and other studies using the same method. Girls were more advanced dentally than boys. Perth boys showed more advanced dental maturation than French-Canadian boys for ages 6-10 years. Perth girls showed advancement over French-Canadian girls for ages 5-11 years. The same pattern was seen with Hong Kong Chinese children. London and Finnish children, however, were more advanced for the older age groups, 10-16 years, compared to Perth children. Dental age as determined from maturity scores using this method revealed a highly significant correlation with chronological age. From our results we conclude that the method devised by Demirjian and Goldstein is accurate and reliable for forensic age determination within the Perth population, but variations do exist between different population groups. PMID- 15335505 TI - Does ethanol enhance cocaine toxicity? AB - OBJECTIVES: Results of in vitro and animal studies suggest that ethanol enhances cocaine toxicity. If so, then that this implies that in ethanol users, postmortem blood cocaine concentrations should be lower, or anatomic evidence demonstrable. METHODS: Drug concentrations and autopsy findings were compared in a sample of 72 accidental deaths, where only cocaine, cocaine metabolites, and ethanol were detected. Findings in ethanol positive (E+) and negative (E-) deaths were compared using multiple Student's t-tests and chi2 testing for categorical variables. RESULTS: There were 47 E-decedents and 24E+. Mean ages were similar (40.2 +/- 8.7 and 37.5 +/- 11.1 years respectively). Mean E was 0.113 (range 0.030-0.350 g/dL), and less than 0.080 g/dL in 50% of the cases. Concentrations of C and BE were not significantly different in E- and E+ groups (C = 1.40 +/- 3.6 mg/L, and 0.76 +/- 1.93 mg/L respectively, BE = 3.04 +/- 5.36 mg/L and BE 2.09 +/- 3.77 mg/L, P = 0.4621 and 0.4520). Organ weights were pathologically increased in both groups, but not significantly different. Body Mass Index (BMI) was less (23.9 vs 25.5), and heart weight was greater (449 vs 407 g) than predicted. Over half the decedents had demonstrable heart disease, and 11% died of brain haemorrhage, though the rate for both disorders was similar in each group. CONCLUSIONS: In two-thirds of the cocaine-related deaths studied, no ethanol was detected. When ethanol was present, no differences between the two groups were identified. The findings suggest that acute cocaine toxicity is not enhanced by ethanol cocaine interactions. However, ethanol concentrations were generally low, and it is possible that increased toxicity is apparent when much larger quantities of alcohol have been consumed. PMID- 15335506 TI - The pattern of homicidal slash/chop injuries: a 10 year retrospective study in University Hospital Kuala Lumpur. AB - Homicides as a result of slash/chop injuries are not commonly reported in forensic literature. A 10 year retrospective study from 1987 to 1996 was performed on the pattern of homicidal slash/chop injuries in University Hospital, Kuala Lumpur. A total of 37 cases were analysed. The ages ranged from 17 to 85 years. The victims consisted of Indonesian (37.8%), Chinese (27.0%), Indian (24.3%) and Malay (8.1%) ethnic groups; 2.7% of the cases were not identified. Most of the cases were due to intentional violence (n = 27), while the rest consisted of domestic violence (n = 2), robbery (n = 2), psychiatric homicide (n = 1), accident (n = 2) and unclassified (n = 3). In the intentional violence group, the majority of the victims (n = 16) had more than five wounds. In contrast, the victims in other categories had less than five wounds each, with the exception of a single case in the psychiatric-homicide group. In homicide victims with a single wound, the most common site of injury was the neck. In those with multiple wounds, the common sites were the head and neck. Sixteen cases showed defence injuries, all of them belonging to the intentional-violence group. The reasons for the high incidence of homicidal slash/chop wounds are discussed, as well as the difficulties associated with interpretation of such wounds. PMID- 15335507 TI - Trends in fatal substance overdose in eastern Saudi Arabia. AB - Two hundred and forty-nine deaths owing to substance overdose were reported to the Medico-Legal Centre, Dammam, Saudi Arabia in the 8 year period from 1990 to 1997. All cases were subject to a full forensic autopsy with toxicological and histological studies. The overall male:female ratio was 7.9:1 and males predominated in all age groups. Saudis outnumbered other nationalities and accounted for 191 (77%) of the cases. Four main categories of overdose fatalities were identified: opiates, prescribed medication, alcohol and carbon monoxide. Opiates accounted for 116 (47%) of all overdose deaths, although such fatal overdoses have been decreasing since 1993. Medically-prescribed drugs accounted for 50 (20%) of the fatalities (21 deaths from antidepressants, 19 from tranquillisers, five from hypnotics, three from paracetamol and two from amphetamines). Ethyl alcohol accounted for 27 deaths (11%) and carbon monoxide for 26 deaths (10.5%). This study shows a sharp fall in overdose fatalities in eastern Saudi Arabia during the last 5 years, which probably reflects the influence of a combined policy of strict legislations against drug trafficking and tight control over prescription of drugs. PMID- 15335508 TI - Pitfalls in the diagnosis of child sexual abuse. AB - Case 1: nurses in charge of a 6-year-old girl in a holiday camp noticed some blood spots on the girl's underwear. The possibility of sexual abuse was considered and the girl alleged that her father was responsible. The father was arrested. A surgeon was asked to examine the girl and planned to do it under general anesthaesia. Meanwhile, the girl was brought to the Forensic Medicine Unit. We found a normal hymen and no detectable anal lesions. The girl complained of dysuria and pollakiuria. Urinalysis revealed the presence of blood, leukocytes, and nitrite. Antibiotic treatment for lower urinary tract infection was initiated and all symptoms improved rapidly. The father was released. Case 2: a 7-year-old boy complained of having been sexually assaulted 24 h before. The boy did not report any pain or bleeding during or after the assault. Examination of the perianal region and of the anal sphineter were normal. Proctoscopy did not show any evidence of trauma to the anal canal. Tests to detect spermatozoa in the rectum were positive. The assailant was arrested. The present cases illustrate that: (i) psychological and social consequences of the biased interpretation of common symptoms may be dramatic in the case of child sexual abuse; (ii) both questioning and examining a child may be difficult for non-specialized practitioners; and (iii) tests to detect spermatozoa should be systematically performed in the case of a suspected or alleged recent assault, even in the absence of any clinical lesions. PMID- 15335509 TI - Suicide notes in the courtroom. AB - The suicide note is of particular interest and importance to the forensic scientist/clinician. In those cases dealing with wrongful death and suicide, the note can shed significant light on other forensic data, just as the other forensic data can shed significant light on the note. This paper presents two forensic cases to illustrate the prima facie (nomothetic) relevance of the suicide note in the courtroom and to address some of its limitations (idiograpic). PMID- 15335510 TI - Trauma and arteriovenous malformation of the brain - a problem in medico-legal practice. AB - Legal difficulties can always arise when a death, owing to a pre-existing natural disease, follows trauma. Arteriovenous malformations of the brain are well known to rupture spontaneously, causing severe and often fatal cerebral haemorrhage. When such rupture is preceded by head injury, it is necessary to consider whether the rupture was coincidental or a direct result of the physical injury sustained. The standard of evidence required for criminal conviction in such cases must be 'beyond any reasonable doubt'. The present article discusses the relationship between physical trauma and cerebral arteriovenous malformation in an adult male who died following an alteration with a colleague that resulted in his being punched in the face, falling down and hitting his head. PMID- 15335511 TI - Drug-mediated sexual assault. AB - Psychoactive substances may be given to a potential victim to alter their behaviour or conscious state. This problem is not new; alcohol has been used alone or in combination with other substances to the same effect. Doctors involved in the examination of complainants of sexual assault need to be aware of the effects of the drugs that may be used and ensure that a detailed history, examination and the appropriate specimens are taken. This report describes a series of incidents involving drug-mediated sexual assault and outlines the dilemmas that accompany such incidents. PMID- 15335512 TI - Recent developments regarding the treatment of persons arrested under the Road Traffic Act. PMID- 15335513 TI - Reproaching television for violence in society: passing the buck? Interim results from a naturalistic study in St Helena, South Atlantic. AB - This article refers to a naturalistic study of broadcast television's effects upon the social behaviour of young pupils in St Helena, South Atlantic. Interim findings showed that 2 years after the availability of television, the good behaviour noted before television arrived had been maintained. Explanations for the continuance of good behaviour focuses upon the attributes of watchfulness and care which Islanders exercise over others. PMID- 15335514 TI - Drug courts and drug treatment. AB - This paper examines the way in which American Drug Courts operate, and the changes required were they to be introduced into the UK. PMID- 15335515 TI - Child sexual abuse: a preliminary study. AB - In 1997, 52 child victims of sexual abuse were examined at Oporto Institute of Forensic Medicine. The authors analysed the data concerning these cases in an attempt to contribute to the elucidation of child sexual abuse, particularly in the north of Portugal. PMID- 15335516 TI - Death as a result of heat stroke in a vehicle: an adult case in winter confirmed with reconstruction and animal experiments. AB - A 54-year-old man was found dead in the driver's seat of his vehicle on a winter's day. Investigations of the vehicle revealed that the engine was running, and the car heater was left on with the maximum temperature and velocity. The body was found excessively sweating. Rectal temperature of the body was 43 degrees C at 10 h post mortem. In autopsy, several superficial skin burns were observed on the face, the shoulders and the legs. The lungs were heavily congested and hemorrhagic. The liver showed typical alcohol-induced micronodular cirrhosis. The alcohol concentrations were 0.17% in the blood of both the left and the right heart, 0.17% in the femoral-vein blood, 0.21% in the bladder urine and 0.34% in the gastric contents. A reconstruction experiment demonstrated that the temperature inside the vehicle rose rapidly and reached 50-58 degrees C in 3 h. Animal experiments showed that the temperature threshold for rats to succumb to heat was between 40 and 45 degrees C. This case shows that heat stroke in a vehicle can occur in adults with chronic diseases or alcoholism, such as in this particular case, even in the winter. PMID- 15335517 TI - Postgraduate education in clinical forensic medicine: a graduate diploma. AB - The absence of a formal postgraduate course has had a significant impact on the development of clinical forensic medicine. Increasing scrutiny by the criminal justice system and recognition of the specialized nature of the work have led to the need for an academic basis for the discipline. Monash University in Victoria, has developed a course leading to a Graduate Diploma in Forensic Medicine. Combining principles and practice taught largely by distance education methods, the programme will enhance practitioners' knowledge and skills in clinical forensic medicine. PMID- 15335518 TI - Homosexual homicides. AB - This report describes four cases of male homosexual homicide committed in Split County, Croatia, in 1991 and 1995. The homosexual orientation of the victims was determined by police investigation. Three middle-aged to elderly person were killed by an unknown offender or offenders, in very similar ways within a few months of end other in 1991. The bodies were found in their houses with multiple blunt force injuries to the head. One victim also had marks of manual strangulation, and was choked with a handkerchief which had been put in his mouth. The last victim was a 45-year-old man, found in the bathroom with 33 stab wounds, mostly in the neck and back, and with two shots in the head from an underwater gun. He was killed by two adolescent males at the beginning of sexual intercourse. The presented cases show a high level of violence with multiple injuries may be associated with male homosexual homicide. PMID- 15335519 TI - False allegations of rape in adults. AB - The term 'false allegation', when applied to rape and assault, is a difficult concept for forensic medical examiners and the police to comprehend. Reliable evidence about how prevalent these allegations are is hard to obtain. This paper is a transcript of the oral presentation given at the 6th Cross-Channel Conference in Clinical Forensic Medicine held in Southampton, May 14-16th, 1998. PMID- 15335520 TI - Silence in interview: psychiatry and the Argent conditions. AB - A man charged with serious sexual offences was advised by his solicitor not to answer the questions which were put to him during the police interview and in the absence of an appropriate adult whose assistance he had declined. At his trial, counsel for the prosecution wanted the jury to be allowed to draw an adverse inference from his silence. Psychiatric evidence was admitted to the effect that he had a paranoid personality disorder. On the basis of the ruling in R v. Argent 1997, in which grounds were set out for not allowing a jury to draw an adverse inference, it was submitted that the jury should not be allowed to draw an adverse influence. These grounds include mental capacity, state of of health, tiredness and personality. The trial judge accepted the submission. PMID- 15335521 TI - Vasovagal death from screwdriver stabbing of the neck. AB - A 40-year-old male victim of an assault with a screwdriver died from vasovagal inhibition as a result of stabbing to the neck. The rare mechanism of death was established after an autopsy. The need to avoid contaminating and adversely modifying clinical forensic evidence is emphasized. A stab wound, by definition, has a smaller skin surface wound but a greater subcutaneous depth of its track. It can pose a clinical challenge for the trauma surgeon in assessing cryptogenic serious internal damage beneath an apparently innocent external wound. In the case reported, severe internal haemorrhage was clinically suspected as there was no significant external bleed from blood vessels in the neck. An autopsy explained the rare mechanism of death. The stab wound in the neck was not explored surgically, but it is pertinent to give a reminder of the need to preserve forensic evidence. PMID- 15335522 TI - Immunodeficiency induced by child abuse. AB - We report the case of a 9-year-old girl who died from sepsis from cellulitis of the neck caused by a right ear injury. The autopsy findings showed severe involution of the thymus and atrophy of lymphoid tissues. The impairment of T- and B-cell functions was demonstrated both histologically and immunohistologically. Thymic involution caused by child abuse might lead to secondary immunodeficiency. PMID- 15335523 TI - Hypoglycaemia: a hidden danger. PMID- 15335524 TI - Re: Robinson SP. Road traffic Act: expert's dilemmas. Journal of clinical forensic medicine 1998; 5(2): 104-106. PMID- 15335525 TI - Re: Robinson SP. Road traffic act: expert's dilemmas. Journal of clinical forensic medicine 1998; 5(2): 104-106. PMID- 15335526 TI - Re: Forensic exhibits-precautions to avoid contamination. PMID- 15335527 TI - Re: Walter H, Samtani A. Misuse of Zoplicone. Journal of clinical forensic medicine 1998; 5(2): 95. PMID- 15335528 TI - Continuing education programme - Victorian Institute of Forensic Medicine and Department of Forensic Medicine, Monash University. AB - Given the increasing requirement of the courts for forensic experts to engage in ongoing education, a continuing education programme was developed in forensic medicine at the Victorian Institute of Forensic Medicine to cater for both clinicians and pathologists. The programme consists of a series of cases which are circulated several times per year. All are actual cases and are reflective of the types of presentations experienced in forensic medicine. Each case includes relevant and appropriate details/findings and may include photographs. A series of questions follow which are answered usually in short-answer format. The answers are returned and correlated by a review panel, and a commentary with the response outcome is distributed to all those involved. The cases presented here are selected from this programme. PMID- 15335529 TI - Judicial applications of hair testing for addicts in Crete: sectional hair analysis of heavy heroin abusers. AB - Laboratory examination of hair from drug users has been employed for the last 5 years in Crete, in addition to the psychiatric or other forensic clinical examinations necessary for the confirmation of a person's use of drugs. The present study reports results of total and sectional head, axillary and pubic hair analysis of imprisoned abusers under interrogation and awaiting presentation at the Crime Court. Morphine levels in total head hair samples 1, 2 and 4 months after preliminary imprisonment ranged from 1.2 to 38.2, 0.5 to 22.5 and 0.1 to 4.9 ng/mg of hair respectively, while the maximum morphine levels amongst all studied groups of those arrested were found in the sections of axillary and pubic hair. Notably high morphine levels in head, axillary and pubic hair sections (13.7, 8.4 and 18.1) ng/mg respectively) were measured, even in abusers who had been imprisoned between 2 and 4 months. Hair tests may considerably assist to evaluate the systematic present and past abuse of heroin and other drugs. Consequently, it may be used as valuable expertise evidence during questioning and in court. PMID- 15335530 TI - The effect of blood in the oral cavity on breath alcohol analysis. AB - The theory that blood (containing alcohol) present in the oral cavity may falsely increase breath analysis recently led to a successful appeal against a drink driving conviction. Subjects who had previously consumed vodka (37.2% alc/vol), at 30 ml/10 kg and reached a BAC (blood alcohol concentration) of between 0.05 and 0.10% were then given four oral solutions consisting of a control (distilled water), and 0.05, 0.10 and 0.15% aqueous alcohol (ethanol) solutions, administered in coded form. A four-way cross-over, blind, randomized assay was conducted with the solutions, with breath analyses conducted in the presence or absence of solution in the mouth. The first trial group (n = 18) received 2 ml of solution, and we found that the simulated 0.15, 0.10 and 0.05% alcohol solutions in the mouth produced BAC reading increases of 0.0088 +/- 0.0014, 0.0062 +/- 0.0008 and 0.0055 +/- 0.0010% respectively (p < 0.001). The second trial group (n = 20) received 1 ml of solution and produced BAC reading increases of 0.0047 +/- 0.0011 (p < 0.001), 0.0023 +/- 0.0008 (p < 0.01) and 0.0020 +/- 0.0006% (p < 0.05) respectively. In conclusion, these studies indicate that small volumes of blood (containing alcohol) in the mouth would not have a practical effect on breath analysis readings. PMID- 15335531 TI - Medico-legal death investigations and autopsies in Istanbul, Turkey. AB - There are two different organizations in forensic medicine in Turkey: the universities and the Council of Forensic Medicine (CFM). The CFM is an official body of the Ministry of Justice and serves as an expert witness on technical and medical subjects. The basic duty of the universities is education in the medical schools with the cooperation of the CFM. This study examines 3064 medico-legal death investigations and 2548 autopsies performed in the CFM, Ministry of Justice, Istanbul and in the eleven units of CFM in Istanbul during 1996. The age and sex distributions, the incidence and causes of deaths were reviewed. PMID- 15335532 TI - Rape in Hong Kong: an overview of current knowledge. AB - This paper summarises all published and unpublished material available on the topic of rape in Hong Kong. Data including epidemiology, victim profile, assault characteristics, are presented. Most of the data shown are from the 15-year period between 1981 and 1995. Gaps in knowledge are highlighted so as to direct the attention of local researchers and overseas research collaborators. PMID- 15335533 TI - Forensic pathological observations on fatal injuries to the brain stem and/or upper cervical spinal cord in traffic accidents. AB - A retrospective study was performed on 149 forensic autopsy cases (12 in Kanazawa and 137 in Munich) of brain stem and/or upper cervical spinal cord (UCSC) injuries as a result of traffic accidents. Pedestrians were the most common type of victims, followed by car drivers. Eleven of the 12 Kanazawa victims were pedestrians. The total ratio of pedestrian victims gradually increased with age. The ponto-medullary junction was the most frequently injured site. A total of 146 cases demonstrated basal skull fractures and/or dislocation of the upper cervical joints: atlanto-occipital joint, atlanto-axial joint, or the joint between the 2nd and 3rd cervical vertebrae. However, in three cases, hyperextension of the neck unexpectedly caused brain stem injury neither associated with basal skull fracture nor with cervical dislocation. Intraventricular haemorrhage was found in 96 cases, suggesting a common feature associated with brain stem and/or UCSC injury. A tear of the basilar artery and that of the carotid artery was found in 17 and 20 cases respectively. A total of 39 victims demonstrated a blood alcohol concentration of over 0.5 mg/ml, and hyperextension of the neck occurred more frequently in these cases. In spite of the marked predominance of immediate death (138 cases), 11 cases (nine of brain stem injury and two of UCSC injury) had unexpected survival times of from 45 min to 12 h. PMID- 15335534 TI - Increasing rate of sexual assaults by adults appearing before the Assize Court of Rennes, France, during 1987-1997. AB - In this paper, we analyse all the sentences handed out by the Assize Court of Rennes in France, in the last decade. A recent increase in the number of sexual offences has been observed. An increase in the number of cases of incest has also been noted. This paper analyses the sex ratio, the age of the victim, the association with other violence and the relationship between the victim and his or her aggressor. The outcomes of the trials are discussed. PMID- 15335535 TI - Death from diesel fumes. AB - A rare fatality from inhalation of diesel motor exhaust fumes is reported and the toxicity of diesel fumes is discussed briefly. PMID- 15335536 TI - Cardiotoxic mechanisms and interrelationships of cocaine: including a single case depicting several of these mechanisms. AB - The underlying mechanisms of myocardial infarction as a result of cocaine abuse appear to be multifactorial. The various cardiotoxic mechanisms and interrelationships of cocaine are fully reviewed, and a chart has been reconstructed to give the reader a clearer understanding of them. Moreover, an unusual case of a 29-year-old male cocaine abuser is presented because it illustrates many of the reported cardiotoxic effects, all of which are present in the same individual. PMID- 15335537 TI - Think drink, think drugs, think overdoses. AB - The police are frequently called upon to deal with people whose behaviour is disturbing the peace of other citizens. A large proportion of such people will have consumed alcohol, some will have consumed drugs and others will be suffering from mental illness. The role of the FME is to determine whether medical or psychiatric intervention is necessary for that individual. Because of the difficulty in assessing a person's mental state when they are believed to be under the influence of substances, it is commonplace for the person to be reassessed some hours after the initial assessment to enable the effect of any consumed substances to abate. However, if the toxic effects of consumed medication are not appreciated, such a delay in receiving medical care may have serious consequences. We report this case as a reminder to all forensic physicians of the clinical effects and possible grave complications associated with amitriptyline overdose. Doctors who work in custodial medicine should be alert to the possibility of an overdose of a drug with anti-muscarinic effects when confronted with a detained person who is acutely confused with clinical features suggestive of stimulant use. PMID- 15335538 TI - Continuing education programme - Victorian Institute of Forensic Medicine. AB - With the increasing requirement of the courts for forensic experts to engage in ongoing education, a continuing education programme (CEP) was developed in the field of clinical forensic medicine at the Victorian Institute of Forensic Medicine in 1996. This programme has been described and was initially established to provide a means of education for the contracted forensic medical officers who provide forensic services to the police via the Institute throughout the State of Victoria. Owing to the sparsity of the population and the considerable distances between forensic practitioners, the CEP was designed to cater for individuals who are working alone: in effect, a distance education programme. Forensic pathologists expressed interest in the programme and it was subsequently modified to include forensic pathology cases. Currently, the programme caters for both clinicians and pathologists, and takes the form of four to five cases with related questions which are circulated several times per year. The cases include a mixture of both challenging and ordinary procedural types that may present to practitioners working in either clinical forensic medicine or forensic pathology, or both. The areas covered include: * injury interpretation * procedural matters in relation to adult and child sexual and physical assault * pharmacology/toxicology interpretation of findings * medico-legal issues (e.g. confidentiality, consent, etc.) * issues relating to alcohol and drugs * traffic medicine * clinical and legal aspects of sudden natural death * suspicious deaths * suicide * interpretation of findings at autopsy * fitness for interview * fitness to plead * psychiatric issues * general clinical medical issues. The presentation of each case includes relevant and appropriate details/findings and may include photographs. A series of questions follow which are answered in either short answer or multiple-choice format. The answers are returned and are correlated by a review panel of forensic physicians and pathologists, and a commentary with the response outcomes is distributed to all those involved. This also includes pertinent references. The cases presented here and in subsequent issues are selected from this programme. PMID- 15335539 TI - Homicide followed by suicide: remorse or revenge? AB - Homicide is followed by the suicide of the assailant in around 4% of homicide suicide episodes in England and Wales. The assailant is invariably a man who most commonly kills his spouse and/or children. Shooting is the most common method of suicide and homicide in these cases. It has been asserted that the low rate of homicide and relatively high rate of suicide in killers is a result of English killers internalizing their culture's abhorrence of killing. However, examination of homicide-suicide episodes indicate that in most episodes the decision to commit suicide has been taken before the decision to kill and that only a minority of suicides in assailants are out of remorse. Homicide followed by suicide is a distinct category of homicide which has features that differ from other forms of killing. These episodes are complex and do not reflect simple remorse following the killing. Homicide-suicide episodes in England are similar to those in countries with higher homicide rates. PMID- 15335540 TI - Who takes alcohol prior to suicide? AB - Alcohol abuse is strongly associated with suicide. Alcoholics are at a high risk of suicide, and studies of case series of suicide show that alcoholics account for between 20% and 40% of all suicides. What is less clear is the role of alcohol in the events leading up to the suicide. This study reviews the characteristics of individuals who consumed alcohol prior to suicide. All cases of suicide assessed by the Department of Forensic Medicine, Dundee University between 1988 and 1995 were reviewed. Data were obtained on blood-alcohol levels of 349 cases, together with the method and circumstances of the suicide, demographic variables and reports of past psychiatric history. Forty-five per cent of suicide cases had consumed alcohol and 19% were drunk (BAC > 150 mg/dl) at the time of the suicide. Consumption of alcohol was not associated with a particular method of suicide, nor with social factors such as employment status, marital status or social class. However, alcohol use was more common among those with no previous psychiatric history. This study confirms that alcohol consumption is a common precursor to suicide. It suggests that alcohol may play a more important role in the events leading to suicide amongst individuals with no previous psychiatric history. PMID- 15335541 TI - Forensic medicine training in Tunisia. AB - Forensic medicine is a specialty new to Tunisia since local teams started working in this field only 3 decades ago. Forensic medicine departments are now established in university hospitals, associated with the four national faculties of medicine. Medical undergraduate education includes forensic medicine as part of the curriculum because of the current lack of forensic physicians and the need for practitioners with some training in this field. Specialization is achieved by a 4-year full-training residency, including both forensic pathology and medicine. The undergraduate and postgraduate education programmes are described. PMID- 15335542 TI - Child death related to insulin omission by mother. AB - A 4-year-old child was found dead in her house. She had been suffering from insulin-dependent diabetes mellitus (IDDM) for 3 years. She had been admitted to hospital three times, and attended the hospital once a month. Her glycaemic control remained poor. Autopsy findings revealed a remarkably fatty liver and lack of beta cells in the pancreatic islets of Langerhans. The laboratory reported ketoacidosis, ketonuria, glycosuria, and high levels of vitreous glucose and ketones. Her father had been absent for the 3 days before her death, and the mother should have fed her. Her mother was found drunk on the floor. No detailed history was available because of the mother's death, but the probability of the mother's omission of the insulin injection was suggested. Poor control of IDDM might have also been closely connected to general neglect by the mother. This case was considered child abuse. PMID- 15335544 TI - Science and politics of cutting and stabbing injuries in the USA. PMID- 15335543 TI - Fatal air embolism occurring during consensual intercourse in a non-pregnant female. AB - Death from air embolism during pregnancy has been reported following sexual activity, particularly vaginal insufflation. We report a death from air embolism in a non-pregnant woman during consensual penile intercourse, in a position with the pelvis elevated above heart level. Air is thought to have entered the veins via a vaginal laceration, which occurred during digital foreplay. PMID- 15335545 TI - Genital pathology: its impact on the interpretation of signs of genital trauma (SEAL development training). AB - A seminar was held in October 1997 at Wood Street Police Station by courtesy of the City of London Police to explore the diagnostic difficulties posed by pathological conditions affecting the vulva and penis, which may cause problems when assessing the nature and degree of genital injury in allegations of sexual assault. While a lot of the presentations were visual, in the form of slides of the discussed conditions, a synopsis of each speaker's contribution is given below. PMID- 15335546 TI - The Audit Commission's report. PMID- 15335547 TI - re: Misuse of zopiclone. PMID- 15335548 TI - Most grateful for your advice. PMID- 15335550 TI - Road traffic act: expert's dilemmas. PMID- 15335551 TI - Medico-legal aspects of doping. AB - Abuse of anabolic steroids is an increasing problem not only among athletes but also body-builders and teenagers. A fast-developing black market has been established since the opening of the borders to eastern Europe. Medico-legal aspects of doping are addressed with particular reference to toxicology and pathology. Constituents of anabolic steroids bought on the black market were identified using gas chromatography/mass spectrometry; the products did not contain the expected ingredients in 35% of cases. Long-term effects and fatalities because of anabolic steroid abuse are reported here based on our own case material and a literature review. In our own cases, severe cardiovascular side-effects developed after long-term abuse of Dianabol (methandrostenolone) and Oral-Turinabol (chlordehydromethyltestosterone), i.e. myocardial infarction, stroke, organomegaly and/or severe atherosclerosis. The pathogenesis of cardiovascular complications (cardiotoxic effect, risk of atherosclerosis, thrombogenic risk) is discussed based on the available literature reports following fatal outcome after the abuse of anabolic steroids. PMID- 15335552 TI - Confirmation of identity by a metallic knee prosthesis in a severely burnt body. AB - Confirmation of identification of the deceased may be obtained in several ways. In this case, identification was confirmed by matching the serial numbers on a prosthetic knee joint and co-existing anatomical abnormalities with the joint serial number and X-ray reports in the hospital notes of the suspected victim. PMID- 15335553 TI - Drug-induced anisocoria in two detainees. AB - Two suspects held by the police were examined for fitness to be interviewed. It was noticed that both had a persistent unilateral dilated pupil in the absence of a history of pre-existing disease or acute head injury. They subsequently admitted to misuse of eye drops. A brief review of the significance of anisocoria (unequal pupils) and drugs that may cause altered pupillary size follows. PMID- 15335554 TI - Self-injurious behaviour in police custody. AB - A Royal Commission into Aboriginal Deaths in Custody in Australia, reported in April 1991, focused attention on the problem of self-injurious behaviour among incarcerated individuals. This paper examines the specific issue of the police custodial setting. A review of the relevant literature has been undertaken to identify risk factors and determine profiles of persons at risk. Preventive approaches are examined and formats for screening questionnaires are advanced. PMID- 15335555 TI - Examination and significance of 'tied up' dead bodies. AB - Examination of dead bodies found with parts of the body tied or bound can pose a real problem for the investigators in that it may not be possible to differentiate between suicide, homicide or accidental death. This article reports five such cases. PMID- 15335556 TI - Summer sandal genital trauma. AB - In this paper four cases are reported of girls presenting with vulval injuries as result of heavy impact against the edge of redundant soles. All four were referred for suspicion of sexual abuse to the Lindisfarne Forensic Paediatric Centre, Newcastle because no obvious mechanism for injury was available from the initial history, and because of apparent delay in reporting of symptoms. In each case, the lack of injury to deeper structures including the hymen, was a strong indicator of their accidental origin. PMID- 15335557 TI - Medical complications associated with the use of rigid handcuffs: a pilot study. AB - The provision of Police Officers with an updated form of rigid handcuffs in 1992 resulted in adverse publicity with regard to the purported 'serious injuries' that these new handcuffs produced and there was a suggestion that they 'may have to be withdrawn'. However, the police appeared to appreciate the ease of application and control of the detained person that these rigid handcuffs afforded. This pilot study documents injuries to the wrist and other sites on 41 detained persons who had been placed in rigid handcuffs in one Metropolitan Police station during a 7-month period. Nearly all of the wrist injuries that were detected were minor and required no specific treatment. Nonetheless, there is a need for further research comparing the medical complications of rigid handcuffs and other handcuffs currently in use. PMID- 15335558 TI - The epidemiology of violence in Denmark. AB - A 1-year prospective study was carried out to describe the epidemiological changes in interpersonal violence in Aarhus (Denmark) over a 12-year period. Data was collected from the Accident & Emergency department, the Department of Forensic Medicine, the local police and compared to previous studies carried out in 1981-1982 and 1987-1988. The incidence of violence was found to have declined significantly over the study period, but severity of violence remained unchanged. The composition of pubs, discotheques and cafes might be an important factor in reducing inner city violence. Police statistics alone may give a false picture of the assault rate. All of these factors are discussed. PMID- 15335559 TI - Breath testing in patients with respiratory disability. AB - The presence of chronic respiratory disease is sometimes claimed as a defence by drivers who refuse or fail a breath test. Trials have been conducted with two breath analysis instruments used in the state of Victoria, Australia in order to determine the respiratory parameters compatible with a satisfactory test. Patients with restrictive and obstructive conditions participated in the trial that was conducted in the respiratory unit of a large teaching hospital. The results showed that the evidential breath analyzer currently used in Victoria was able to accept breath samples from subjects at the limit of respiratory function likely to be encountered in drivers. It was also found that a subject capable of providing a breath sample for the current screening device should also be able to provide an evidential sample. The trials provided useful information in evaluating defences of inability to provide a breath sample due to respiratory incapacity. PMID- 15335560 TI - Deaths in police custody during 1994: a retrospective analysis. AB - This paper presents the results of a retrospective analysis of deaths in police custody from 1 January to 31 December 1994. During that year, there were 32 such deaths in the UK. For purposes of analysis, the deaths were grouped into three principal categories based on the frequency of their occurrence: deaths by hanging (12); deaths among those arrested for drunkenness (11) and other deaths (9). By considering the results of this study together with a comprehensive literature review, several strategies for reducing the frequency of deaths in police custody are proposed. PMID- 15335561 TI - The changing status of forensic medicine in Greece. AB - The practice of forensic medicine in Greece is similar to that in other European countries. That is Greece follows the continental model of sudden death investigations, whose norms govern the exercise of forensic medicine through Greek criminal procedure law. In Greece, a university and Government system of forensic medicine exist and occasionally work side-by-side. University departments are strategically placed in all districts of the country. State forensic medicine services are located in Athens, Piraeus and Crete and function in parallel with the local University departments of forensic medicine. Forensic examinations on the living are also exclusively performed by the University and State Legal Medicine services. The cases are simply divided according to the countries of these areas. A recent regulation by the Supreme Court eliminated the overlapping of jurisdictions emphasizing the role of the University departments, and finally establishing a new University-oriented medico-legal system throughout Greece. PMID- 15335562 TI - The influence of the media on the incidence of violence. AB - There has been much debate as to whether the media have a direct influence on the incidence of interpersonal violence and violent behaviour. The producers of cinema and television programmers and newspaper editors deny this influence and maintain that they merely reflect the current existence of violence in society. PMID- 15335563 TI - Stalkers and other obsessional types: a review and forensic psychological typology of those who stalk. AB - Stalking, an age-old phenomenon, was discussed in the ancient writings of Hippocrates, researched extensively in the early 20th century by French psychiatrist G.G. de Clerambault, given notoriety with the tragic death of former Beatle John Lennon, and viewed as 'Fatal Attraction' in local theaters across the world. Stalking is unique as the repetitive, harassing, long-term course of conduct and behaviour reflects the internal dynamics of the stalker's motivating mental disorder. Furthermore, the investigation of stalking is difficult. The numerous and progressive acts, comprising the intentional course of conduct, require a continuity of purpose: harassment designed to control the victim. A multi-disciplinary approach, utilizing law enforcement, legal, medical and mental health professionals, is necessary to effectively eradicate this encroaching behaviour. In 1990, the first Anti-Stalking legislation was passed in California, USA. In just 6 years, all 50 states have followed suit. Public awareness has encouraged federal legislation to stop stalking behaviour as a course of conduct crime and to give victims a measure of protection. This research paper overviews periodicals, law review articles, studies and projects of law enforcement, legal counsel, medical professionals, forensic mental health professionals, forensic examiners and criminologists since 1992. The study of stalkers and stalking behaviour is a work in progress. As questions are answered, statutes are amended and diagnoses are distinguished, it becomes a stepping stone to the future understanding of the pervasive stalking phenomenon facing our culture and society today. PMID- 15335564 TI - The Cinderella section: room for improvement in the documentation and implementation of Section 136 of the Mental Health Act 1983. AB - A questionnaire sent to all 43 police forces in England and Wales confirmed that there was a lack of understanding of the documentation issued by central government detailing section 136 of the Mental Health Act 1983. A survey of forensic physicians in England and Wales reveal that doctors providing medical care in police stations had insufficient knowledge of the Act. PMID- 15335565 TI - Fatal compressional injuries from a burst tyre: an industrial death. AB - A young maintenance man was killed while fixing a heavy duty tyre to a fork lift. The tyre exploded, sprang off the front axle and threw the victim against a wall 7.5 m away, fatally crushing him. Autopsy revealed multiple crush injuries and, cannabinoids were detected in the victim's urine. The mechanism of the fatal pneumatically-induced compressional injuries sustained and the significance of the cannabinoids in the body fluids of the deceased are discussed in this paper. Proposals are given on how to prevent similar accidents in the workplace. PMID- 15335566 TI - Health care and the prisoner: a human rights perspective. PMID- 15335567 TI - Drugs and testamentary capacity. AB - Drugs, or withdrawal from them, can cause changes in consciousness, reasoning ability, perception of reality, and memory, all of which could affect testamentary capacity. When asked whether a person is, or was, capable of making a will, one should take into account the possible effects of drugs or drug withdrawal on the person's ability to comprehend the nature of a will, the assets to be disposed of, and the potential beneficiaries. PMID- 15335568 TI - Satanic abuse, with focus on the situation in Finland. AB - This paper outlines Satanism and devil worship as practised in the Western countries and reviews the occurrence of Satanism in Finland. Two principal groups can be distinguished: the Satanists, mainly adults embracing the philosophical aspects of Satanism with no interest in hurting others, and the devil worshippers of Satanic cults, who accept teenagers into their group and whose activity may take violent forms. The main Satanic cult activity is vandalism, but other activities are now becoming more aggressive: causing bodily and mental harm to members and victims and luring young people into criminal activity. The views of the police and the medical community are discussed in this paper and current intervention is examined. PMID- 15335569 TI - Sudden unexpected deaths from ruptured abdominal aortic aneurysms. PMID- 15335570 TI - Continuing education in forensic medicine: an exercise in distance learning. AB - Distance education and its application to postgraduate clinical medicine and continuing education in medicine is a recent development. Given the increasing requirement of the courts for forensic experts to engage in ongoing education, a continuing education programme (CEP) was developed in the field of clinical forensic medicine. This programme is the first programme available in clinical forensic medicine. Due to the interest expressed by forensic pathologists, the CEP has been modified to include cases of forensic pathology interest and the programme is now subscribed to by both clinicians and pathologists. The programme is reflective of the types of cases experienced in forensic medicine and is designed for individual practitioners working alone. PMID- 15335571 TI - Lesions that simulate gunshot wounds. AB - A variety of postmortem and antemortem defects can be misinterpreted as true antemortem injuries. In this paper, we present a collection of six cases with defects which simulate gunshot wounds. We briefly discuss each case and emphasize the importance of scene investigation, clinical correlation and careful examination in such cases. PMID- 15335572 TI - Eight cases of suicide by self-cutting or -stabbing: consideration from medico legal viewpoints of differentiation between suicide and homicide. AB - The present article describes eight cases of suicide by self-cutting or stabbing. Seven of the deceased had a history of psychiatric problems. In every case, sharp weapon injuries were observed at typical sites of self-inflicted injury. The self-inflicted injuries varied in severity from a single stab wound to multiple cut wounds. In four cases, hesitation marks were found. Among the remaining four cases without hesitation marks, only one had clothing injuries. Based on the scene investigation by police, it was deduced that each body had been found at or very near the place where the fatal injuries had been sustained. In two cases, toxicological analysis played an important role in discriminating suicide from homicide. In one case, a 52-year-old male stabbed himself fatally after potassium hydroxide ingestion, and in another, a 71-year-old male committed suicide by self-cutting and -stabbing after drinking an alkaline detergent solution. In five of the eight cases, unusual findings such as a single stab wound, multiple cut injuries, no hesitation mark or presence of clothing injuries were found. In these five cases, the psychiatric history of the deceased contributed to furthering our comprehension of these unusual findings. We believe that thorough toxicological analysis and scene investigation as well as autopsy are essential in discriminating between homicide and suicide. Furthermore, forensic pathologists and forensic physicians at the scene should obtain as much information as possible on the victim's previous character including previous psychiatric history in order to elucidate his or her manner of death. PMID- 15335573 TI - Fatal and non-fatal outcome by accidental intoxication with paint thinner. AB - Although poisonings (fatal and non-fatal) due to intentional or accidental acute exposure to toluene or toluene mixture fumes have been previously reported in the literature, several issues concerning lethal doses or lasting post-exposure neuropathological impairments still remain unclear. Two male painters (18 and 30 years old) were accidentally exposed to toxic concentrations of paint diluent fumes containing toluene (TL), acetone (ACT) and methyl ethyl ketone (MEK) (60:15:15 w/w/w respectively) during their work in an underground reservoir. Both workers were found unresponsive by colleagues and were immediately transferred to the regional hospital. On admission, the younger man was pronounced dead, while the other remained in the intensive care unit for 3 days and then 4 days in the internal medicine ward. TL, ACT and MEK concentrations in blood samples taken from the survivor on admission were 6.3, 30.6 and 40.5 microg/mL. Postmortem toxicology of the dead worker revealed TL, ACT and MEK blood levels of 12.4, 90.8 and 80.4 microg/mL respectively. The solvent levels in the liver, kidney, lung, brain, testis and gland were also quantified and showed a somewhat similar distribution of the chemicals among these tissues with the highest levels found in the brain and the liver. The fatal and the non-fatal outcome that resulted despite similar intoxication conditions, most probably demonstrates the interindividual tolerance among the painters who also had similar body weights. The surviving painter did not develop any neuropsychological impairment in post exposure time. The reported case strongly emphasizes the necessity to take precautions when using paint diluents in enclosed spaces. PMID- 15335574 TI - Management of potentially violent detainees. AB - Most doctors and other health care workers have at some time received threats of violence, but the assessment of police detainees can be especially challenging and in a recent study of police surgeons, 18% had experienced physical violence. Aggression towards carers rarely erupts without warning and there is much that the examining doctor can do to minimize personal risk. This review offers a brief overview of current understanding about the causes of violence and offers guidance on the systematic assessment of risk and how to manage this within the setting of a clinical assessment of a detained individual. PMID- 15335575 TI - Future of education in clinical forensic medicine: summary of multidisciplinary forum. PMID- 15335576 TI - Some police surgeons around the UK permit the administration of methadone intravenously under their supervision in custody centres. PMID- 15335577 TI - Not licensed for a particular route of administration. PMID- 15335578 TI - Methadone is not licensed for intravenous use. PMID- 15335579 TI - Little I can add to the comprehensive coverage of this subject. PMID- 15335580 TI - Domestic violence: do forensic physicians have a role? AB - The role of the forensic physician in domestic violence incidents in South London was investigated over a 6-month period. The aim of the study was to identify the nature of domestic violence, to determine whether the individuals involved had previously been in contact with the primary health care team or other agencies and whether direct communication between the examining forensic physician and the patient's general practitioner was of assistance to the latter. We concluded that there is a definite role for the forensic physician in documenting injuries in victims and assailants (although most were minor injuries), providing appropriate medical treatment, supplying information regarding support agencies and passing the relevant information, with the patient's consent, to the primary health care team. PMID- 15335581 TI - Medico-legal considerations of deaths from watersports among Caribbean tourists. AB - Watersports are the dominant recreational sporting activity in the Caribbean islands. The economy of many of these islands is tourist-oriented and millions of tourists take part in such sports annually with some fatalities. A total of 13 watersport-related deaths observed in the Cayman Islands over a 2-year period are presented. Complete autopsies with toxicological and histological studies were carried out in each case. Preexisting cardiovascular disease such as hypertension, coronary artery disease (or relative insufficiency) and cardiomyopathies were observed to be significant risk factors and the mean age at death was 53.7 years. Sudden cardiac deaths predominate. One fatal jet skiing accident also occurred. Tourists are advised to undergo complete medical investigations (including ECG and stress tests) prior to engaging in watersports. Drugs of abuse are to be avoided for medical reasons and insurance considerations. Watersports operators must ensure that their clients complete a medical checklist. All equipment and accessories must be functioning optimally to avoid accidents contributed to by such negligence. Consideration should be given to introducing medical tests prior to signing on their clients and there must be adequate supervision. Some form of governmental regulation and enforceable supervision is necessary. PMID- 15335582 TI - Child abuse examination enquiry. AB - OBJECTIVE: To determine how police surgeons in the United Kingdom performed examinations of child abuse victims because wide variations in protocols and procedures have been reported. METHOD: Practising members of the Association of Police Surgeons were requested to complete an enquiry form. Replies were anonymous and subjects covered included gender, work area, training, procedures and protocols for joint examinations with paediatricians and the use of digital or instrument examinations in alleged penetrative sexual abuse. RESULTS: Of all those eligible, 47.7% had responded. Males made up 74.7% of respondents. Published guidelines were followed by 85.3% and 69.8% used purpose-built examination suites while 8.8% still used police station facilities. Procedures for joint examinations were working satisfactorily, according to 79.2% of respondents who also considered that the forensic physician should take the lead role in examinations. Clinical findings were recorded on a prepared aide memoire by 35.2%. Colposcopes were used by 3.4% while 23.3% of respondents considered that a colposcope should be used but cost was an inhibiting factor. CONCLUSIONS: Reported variations in procedures and protocols in examining child abuse victims were confirmed. More discussion and liaison between agencies concerned is required and a firm commitment to the paramountcy of the child's interests is needed. PMID- 15335583 TI - A study on patients presenting with injuries to the eye. AB - Ocular injuries are a common occurrence, as the eye is very susceptible to trauma. This study was carried out to determine the main causes of injuries to the eye and to assess the degree of impairment of vision caused by such injuries. A total of 191 victims of ocular injuries who presented for medico-legal examination during a 1-year period were analysed in respect of their sex, age, type of injury and degree of visual impairment. The majority of victims in this study group had sustained injuries following assault. There did not appear to be any correlation between use of a weapon and the gravity of the eye injury. Victims with acid burns although fewer in number, had a higher proportion with a more severe degree of visual impairment. PMID- 15335584 TI - DNA typing from stained sperm-positive vaginal smears: four rape cases. AB - DNA typing from stained sperm-positive vaginal smears was performed in four cases of sexual assault. In Cases 1 and 2, a 6-month-old and a 6-week-old, Papanicolaou stained vaginal smear with 99% propanol fixation underwent DNA analysis respectively. In Case 1, the HLA-DQalpha type of the sperm fraction of the smear was able to be determined following PCR, and the possibility of the suspect being the rapist could not be excluded. In Case 2, the possibility that the suspect was the rapist in the crime was excluded based on the results of DNA typing by means of two different single-locus probes. In the remaining two cases, H-E-stained smears after air drying for 4 or 5 months were the material for DNA analysis, and DNA typing was attempted by PCR-based STR system. In Case 3, there was no genetic information from sperm fraction of the smear due to the small number of spermatozoa on the glass slide. Therefore, it was impossible to determine whether the suspect was the rapist or not. In Case 4, sperm DNA from the smear was amplified and typed, and the suspect could be excluded. These four cases demonstrate that stained vaginal smears containing many spermatozoa can be used for DNA analysis in sexual assault cases. PMID- 15335585 TI - Myxoedematous madness and grievous bodily harm. AB - Psychosis is a rare but recognized complication of hypothyroidism (otherwise known as 'myxoedematous madness'). This is the report of a man charged with attempted murder who was found to be suffering from myxoedematous madness. The case illustrates several problems in the law relating to mental disorder and criminal responsibility. He was interviewed in the absence of an appropriate adult and this led to the inadmissibility of the police interviews. However, to have laid the basis for a defence of insanity the ideal evidence would have been what he told the investigating police officers about his state of mind at the material time, under caution and possibly with the assistance of an appropriate adult. As it was, although he was clearly mentally ill at the material time, the insanity defence was not open to him as he did not satisfy the MacNaughton Rules and his mental state did not fulfill the criteria for automatism. By the time he stood trial he was fully recovered from his psychosis and this faced the judge with a difficulty in sentencing him. The case is used to discuss proposed changes to the law concerning insanity. PMID- 15335586 TI - Suicide by alcohol overdose. AB - Alcohol is the most often detected substance in the body of a person who has committed suicide. It may be used to reduce the last instinctive hestitations to taking one's own life. Suicide is common in cancer patients. People with cancer related depression often use alcohol as a means of coping but they very rarely use it in order to kill themselves through acute intoxication. However, a case of a cancer patient who committed suicide consuming two bottles of spirit was recently investigated and the conclusions are presented. The post-mortem cardiac blood and vitreous humor alcohol level was found to be 9.0 and 6.2 mg/ml respectively. PMID- 15335587 TI - Balancing the specialist forensic medical witness. AB - This paper examines the role of forensic medical examiners as expert witnesses and advances the argument for them to incorporate more defence perspectives in their work, so that they can give a better balance of responses. Three case studies are presented and these illustrate defence aspects and enable defence orientated references to be cited. The paper concludes with comment on the strength of the evidence that a forensic medical examiner can give when acting for the defence. PMID- 15335588 TI - Re: Dowry Cannibalism. Husain M, Rizvi S J, Usmani J A. Journal of Clinical Forensic Medicine. 1996; 3: 179-181. PMID- 15335590 TI - The human body as an electric circuit. AB - This is a review of the effects of electricity on the human body with an emphasis on the clinical effects which may be of forensic interest, both in the living and after death. The review will explore the interaction between electricity and the human body in its functional and pathological aspects. Electricity interacts with the body in many ways ranging from a manifestation of normal physiological activity to a cause of severe tissue injury and death. Its mode of action and the pathogenesis of injuries caused by electricity are sometimes difficult to understand because it is invisible and intangible. PMID- 15335591 TI - Identification of human remains: forensic radiology. AB - The identification of human remains is of paramount importance in medicolegal investigations. The comparison of antemortem and postmortem radiographic records is one of the main techniques used to achieve a positive identification. The purpose of this paper is to present a comprehensive review of the use of roentgenography in forensic medicine with special emphasis on the nature of the radiographic markers often utilized for identification. Clinical radiologists should be aware of the importance of storing radiographs over prolonged periods of time and of efficient record keeping methods to enable prompt retrieval of X ray films in mass disaster situations. Furthermore, because of his skills in radiographic evaluation, the radiologist's expertise might prove invaluable in forensic consultations. PMID- 15335592 TI - Alcohol misuse in clinical forensic medicine. PMID- 15335593 TI - Volatile substances and their potential to interfere with breath alcohol reading instruments. AB - Two cases are presented which highlight the potential for abused volatile substances to interfere with the results of evidential breath testing equipment. Home Office guidance, issued in 1983, advises that breath testing devices should not be used when a driver is suspected of glue-sniffling, but the advice does not appear to be widely known. The implications for both police officers and police surgeons are discussed. New evidential breath testing devices, which are planned to be introduced into the UK in the first half of 1997, are designed to recognize interfering substances such as solvents. The potential problems these new devices may pose are considered. PMID- 15335594 TI - Medical evidence concerning the suitability to succeed to the tenancy of a farm: the case of 'toad of toad hall'. AB - A hill farmer applied to succeed to the tenancy of his farm on the death of his mother who had leased the farm from his maternal aunt. As landlord, she opposed the application which was heard at an Agricultural Land Tribunal (ALT) convened under the provisions of the Agricultural Holdings Act, 1986. The ALT is a court of first instance. The applicant had to prove his eligibility and his suitability. His eligibility was not disputed. Both parties called medical evidence having a bearing on his physical health which was a matter relevant to the issue of suitability. It was in question because there were allegations of excessive drinking and alcohol-related offending. Finding no evidence that the applicant's drinking affected his farming ability, the Tribunal found in his favour and his application to succeed to the tenancy was granted. PMID- 15335595 TI - Beware of charm needles! AB - This report describes the case of a man accused of murder who had the prosecution case dismissed after he complained that he had been coerced (following assault with a needle) into providing a statement. Subsequent events determined that a more complete examination following his complaint may not have resulted in the case being (wrongfully) dismissed. PMID- 15335596 TI - The use of CS spray in the mentally ill. AB - CS sprays are now being widely used by police in the UK. Concerns are being expressed regarding the toxicity of this substance and some of the situations in which it is being used. This is the first reported case in the UK of CS spray being used to restrain a mentally ill person in police custody. It raises important issues regarding the welfare of mentally ill people who happen to find themselves in contact with the police. There is a need for open debate, specific guidelines and a system of monitoring the use of CS in such situations. PMID- 15335597 TI - Undetected ligature self-strangulation: disadvantage at the scene of a decomposed body. AB - A case of ligature self-suspension which remained undetected until the body was properly examined in the mortuary is reported. It emphasizes the need for the police surgeon attending the scene to undertake careful observation at the scene of death, particularly when postmortem decomposition is present. PMID- 15335599 TI - re: Airomir Inhaler, produced by 3M Healthcare Ltd - possible influence on breath alcohol testing for road traffic law enforcement purposes. PMID- 15335598 TI - re: Dutch and Scottish clinical forensic medicine: a comparison. Turnbull J A, Hannaford P F, Busuttil A. 3: 15-19. PMID- 15335601 TI - Is the nasopharynx warmer in children than in adults? AB - Recent studies on the aetiology of the Sudden Infant Death Syndrome (SIDS) have suggested that some of these deaths are the consequence of an overwhelming inflammatory response to the production of pyrogenic toxins from bacteria colonizing the upper respiratory tract, particularly the nasopharynx. The pyrogenic toxins of Staphlococcus aureus, one of the likelier bacterial candidates, are only produced in temperatures of over 37 degrees C. This study examined nasopharyngeal temperatures in children. It is a preliminary study to develop an accurate means to measure how close to 37 degrees C the nasopharyngeal temperature lies in infants at the age when SIDS deaths occur. Following a pilot study and power calculation, measurements of nasopharyngeal temperature were made on 30 apyrexial children aged 4-10 years and 30 adults with no nasal pathology, undergoing surgery under general anaesthesia, using an accurately sited thermocouple probe. The mean temperature in children (35.64 degrees C) was significantly higher than in adults (34.01 degrees C). Comparable measurements attempted with the same subjects awake gave similar results. PMID- 15335602 TI - Murder: the importance of the structural and cultural conditions of society. AB - This paper argues that a crucial feature in studying murder is to recognize that the structural and cultural conditions of a society may provide the context in which psychopathologies are created and emerge to such devastating effect. The paper focuses on some recent contributions in the development of a widening academic interest in the phenomenon of serial killing, in particular, and then considers murder in general. Issues of time and place are paramount and lead to two types of question: Firstly, is serial killing quintessentially a modern phenomenon? Secondly, why does it occur in some countries and not others? The discussion then considers why there is traditionally a low murder rate in England and Wales. The recent works of Bernard Capp, Maria Tatar and Elliott Leyton provide the framework for trying to answer these questions. PMID- 15335603 TI - Substance-misuse: the front line at the police station - an opportunity for intervention. AB - The Brighton project, 'Get It While You Can', has two ex-drug addict project workers visiting prisoners in police custody cells. They provide information about harm reduction and service provision within the Brighton area. They are also able to provide on-going support to the detainees after they have left the police station and to help them successfully access agencies in the town and further afield. PMID- 15335604 TI - Blood or needle phobia as a defence under the Road Traffic Act 1988. AB - A man was charged with driving over the prescribed alcohol limit. Both specimens of breath contained less than 50 micrograms of alcohol per 100 ml and he refused to submit to a blood test. His defence was that he had a phobia of blood and that he should have been allowed to claim to replace the breath specimens with a specimen of urine. Evidence that he had a phobia was accepted by the prosecution. A woman was arrested on suspicion of driving whilst unfit through drink or drugs. She was charged with refusing to provide a specimen of blood. It was her defence that she had a phobia of needles. Evidence that she had a phobia was not accepted by the prosecution. In both cases the court was asked to decide whether or not the forensic medical examiner had been seriously wrong in deciding that there was no medical reason for refusing a specimen of blood. The man was acquitted and the woman was found guilty. These cases are used to describe the law relating to blood or needle phobias and to suggest how such cases should be approached by the police and forensic medical examiners. PMID- 15335605 TI - Dowry cannibalism. AB - The term dowry cannibalism is used in this article to describe a unique case where the motive for cannibalism was the non-fulfilment of a husband's dowry demands from his wife. The husband was accused by the wife of regularly using a syringe to withdraw blood from her, with the intent of causing her death, and drinking it. Intriguing as this is, it is also medicolegally important: this method of killing might leave no trace suggesting murder. It is postulated that this form of abuse may also significantly alter the psychological state of the victim and hasten death. The case is not yet concluded, and may never be, thus details to allow identification have been omitted from the report. PMID- 15335606 TI - Fatal methamphetamine poisoning in police custody. AB - A fatal case of methamphetamine poisoning whilst in police custody is described. Drug identification and quantification in postmortem blood, urine, brain and gastric contents was achieved using gas chromatography-mass spectrometry. The blood concentration of methamphetamine was 4.92 micromol/100 g, which is in the lethal range. It is necessary for police officers to have a good knowledge of the toxic effects of abused drugs. PMID- 15335607 TI - Hazardous infant and early childhood sleeping environments and death scene examination. AB - Infants and young children are at risk of accidental asphyxia from unsafe sleeping environments Specific dangers include wedging/entrapment and positional asphyxia when infants slide into gaps between mattresses and cot sides or between beds and walls. Faulty cot construction may result in defects through which infants may fall and become suspended, or in spaces which entrap the head. Projections on the inside of cots may ensnare clothing causing hanging, as may nearby curtain cords and restraining harnesses. The mouth and nose may be covered by plastic sheeting or by the mattress in partially filled waterbeds. Rocking cradles with excessive angles of tilt may also place infants' heads into positions where breathing is compromised. Finally, external pressure by adults in shared sleeping situations may result in fatal chest compression or airway occlusion. In many of these situations the autopsy findings are entirely nonspecific and do not help in establishing a cause of death. Thus, careful death scene examination with later formal review of findings may be the only way to accurately determine the manner of death. PMID- 15335608 TI - Traumatic rupture of the aorta. AB - The aim of this study was to determine the incidence and mortality from traumatic ruptures of thoracic aortas (TAR) in a well defined population over a 7 year period. The study provides a retrospective analysis of case files of all deaths from road traffic accidents and falls from heights, the usual causes of TAR, filed in the Forensic Medicine Unit of the Pathology Department of the University of Edinburgh, for Lothian and Borders Region. Of the 276 fatal road traffic collisions and falls from heights identified in archival material, 66 traumatic aortic ruptures were found in the period between 1988 and 1994. Only 11% of these patients were alive on arrival to a hospital or an emergency department. To decrease the high mortality rate the following are required: an earlier, more sustained and vigorous, on site resuscitation; efficient screening methods for the diagnosis of TAR in multi-traumatised patients; identification of the specific characteristics of those traumatic episodes which are likely to be associated with a high risk of rupture, and an increase in the number of surgical interventions for ruptured aortas. PMID- 15335609 TI - Genital trauma in young boys. AB - The issues surrounding genital trauma to young boys has received little attention in recent literature. Five cases of genital trauma are described. The cases demonstrate a number of similar features in their injury pattern. Analysis of the wound pattern will usually assist in defining the injurious force. The circumstances of causation, in particular the non-sexual nature of the genital assault, warrant careful investigation and assessment. PMID- 15335610 TI - Asthma management in police study. AB - Asthma is a common condition and, therefore, may be seen by the clinical forensic physician and police in widely different circumstances ranging from the autopsy room to roadside breath alcohol testing. This paper concentrates on the assessment and treatment of asthmatics detained in custody and does not refer to the Road Traffic Act provisions. A careful history and objective recording of simple severity markers and peak expiratory flow rate should identify patients who may require hospitalization or urgent treatment. Asthma mortality may be reduced by better understanding of the disease and its treatment and by greater use of prophylactic rather than reliance on relief therapy for bronchospasm. PMID- 15335611 TI - Homicide with a screwdriver and simulation of a similar offence by self infliction of injuries. AB - In an underground garage of a department store, a 52-year-old woman was attacked and fatally wounded by several stabs with a screwdriver. The offence happened as the woman was getting into her car. The homicide itself and the subsequent trial were extensively covered in the local media. A few days before the judgement was pronounced, another attack in an underground garage was reported to the police of the town, where the homicide had happened. A 52-year-old woman claimed to have been hurt by an assailant. The wound pattern (several equally shallow cuts arranged in groups), however, suggested self-infliction, which was finally admitted by the informant. The 'victim' used a newly bought kitchen knife to cut herself and left it at the place of the alleged assault. The knife blade was soiled with blood. Referring to the pertinent literature both cases are compared with regard to the injury characteristics and to the damage to the clothing. The morphological features of screwdriver injuries, the criteria differentiating self infliction and assault as well as the manifestation of copycat offences are discussed. PMID- 15335612 TI - The use of hair analysis for the detection of drugs: on overview. PMID- 15335613 TI - Recovered memories of childhood sexual abuse: a concise social history of the phenomenon, and the key psychological concepts relevant to understanding the disputes concerning such claims. AB - The alleged and disputed repression of serious, traumatic and repeated childhood sexual abuse memories, and their recovery, usually with psychotherapy and 'memory recovery techniques', have become a major social phenomenon in the last 15 years. This review picks out the key historical, theoretical, research and legal milestones during that period. Key concepts and a review of the inconsistent usage of language and definitions are highlighted to assist the reader to understand the nature of the dispute. A comparison is drawn between this phenomenon and the parallel social history of claims of alien abductions in the US for the purpose of hybrid breeding. A comparison is also made between the boundaries of natural memory functioning as currently understood, and that imputed to questionable recovered memories of childhood sexual assault. PMID- 15335614 TI - Suicide by fire in Scotland: 1980-1990. AB - Suicide by fire is relatively rare in the Western world, accounting for approximately 1% of completed suicides by all methods. This survey identified suicidal deaths among fatalities known to the Scottish Fire Brigades as having died in a fire incident during 1980 to 1990. It is accepted that this is just one possible definition of what constitutes a suicide by fire. Forty cases were identified as suicides from the circumstances of the death as narrated in the police and Fire Brigade reports of the incident. Autopsy and toxicological reports were examined for all cases and the characteristics of the victim and the incident are described. Most cases occurred in the home of the victim and three quarters of the victims were pronounced dead at the scene or when first seen by a doctor. Smoke inhalation was the most frequently certified cause of death. Alcohol was present in less than half the cases and toxicological analysis for drugs was performed in the minority of cases. One-quarter of victims were reported to have made at least one suicide attempt previously. A range of apparently precipitating factors was observed: the number of cases which appeared to be impulsive responses to life events suggests that prevention is difficult. PMID- 15335615 TI - The Singapore naltrexone community-based project for heroin addicts compared with drugfree community-based program: the first cohort. AB - The cornerstone of heroin rehabilitation in Singapore from 1991 to 1993 was the Three Phase Program (3PP). This was a multimodal approach embracing job placement, individual and family counselling, home placement and restriction of movement by electronic monitoring. Opioid addicts in Singapore are detained by executive order. Inmates volunteer to be on the community program. The completion rate (i.e. the number of inmates presumptively drug-free after 1 year) was 24%. In August 1993, naltrexone, an opiate antagonist without euphorigenic properties was added to the 3PP creating the Naltrexone Pilot Project (NPP). Sufficient similarity existed between the 3PP and NPP for some comparison between the two. The missing link was the chemical blockade offered by naltrexone to provide a deconditioning experience for the decay of learned responses to heroin. The subject can confidently be exposed to drug cues without fear of relapse. Subjects on naltrexone for the first year had a completion rate of 76.3%. In the second year without naltrexone, electronic tagging and counselling, 32.2% were still drug-free. The subject type who was successful was married, older and better educated with late onset in heroin abuse. PMID- 15335616 TI - A new hypothesis for sudden infant death syndrome: the occlusion of vertebral arteries as a major cause. AB - Many factors, such as prematurity, low weight and minor infection, may predispose to Sudden Infant Death Syndrome (SIDS) but not cause it. It is suggested that the ultimate act of dying is due to the occlusion of one or both vertebral arteries triggered by the position of the head and cervical spine. When the head is turned to one side, the ipsilateral vertebral artery may be occluded at mid-cervical level along its length, or the contra lateral artery at atlanto-axial level or on both sides with the head in the initial position. Extension of cervical spine, with or without rotation, may produce obstruction of one or both vertebral arteries. If the degree, the levels and duration of occlusion are exceeded, particularly if there is a degree of failure of the collateral circulation (including one from carotid arteries via posterior communicating arteries), this can lead to prolonged apnoea, a 'near-miss' or cot death. The factors which may predispose to SIDS among others are hypoplasia or 'unexercised' vertebral arteries (in utero) and/or hypermobility of the immature cervical spine with lax joint capsules and ligaments. These factors allow the undue stretch or kinking of the vertebral arteries resulting in occlusion when the head or cervical spine are turned. The hypothesis explains the reduction of mortality of SIDS in infants in sleeping supine position to those in the prone position. In prone position, the infant's head is rotated 90 degrees which may put stress on atlas and axis thus occlude vertebral arteries by stretching and kinking. Infants under 6 months are unable to change position unaided so duration of obstruction is an aggravating element. These factors are about 50% less in the supine position. PMID- 15335617 TI - Passive inhalation of cannabis smoke: a novel defence strategy? AB - Defence lawyers sometimes argue that the presence of cannabinoid metabolites in the defendant's blood or urine resulted from passive unintentional inhalation of environmental cannabis smoke. It would be useful to be able to differentiate passive inhalation from active use so as to discourage the potential abuse of this phenomenon by the defence. Four cases from two jurisdictions in which passive cannabis smoking was used as a defence are presented to illustrate this dilemma. It remains impossible to define objectively an upper limit for blood and urine levels in cases of passive inhalation of cannabis from the environment. However, a claim of passive inhalation, or indeed 'deliberate passive exposure' could be discouraged by making it an offence to place oneself in a position of being 'concerned' in the use of the drug. The onus should be on the defendant to prove that he had not attempted to extricate himself from the situation, being aware of the smoking of cannabis in his immediate vicinity; ignorance would not be an excuse. PMID- 15335618 TI - A case of deferred death from drowning of a man found dead in bed at home. AB - An autopsy case of deferred death from drowning is reported. A 63-year-old man was found dead wearing wet clothing in his bed at home. At a medico-legal autopsy, mud was observed in the air passages as well as in the stomach, and a total of 210 g pleural effusion was also noted in the thoracic cavity (left 70 g, right 140 g). The left and right lungs were expanded and weighed 800 g and 950 g, respectively. Histological examination revealed the pathological finding of pneumonia with the exudation of numerous leucocytes into the alveoli as well as some postmortem changes. Diatoms were detected in the liver and both lungs by the disorganization method. In addition, scene investigation disclosed that he had fallen into a ditch in front of his house and had inhaled muddy water in the ditch, and had then managed to reach his bed and survive for a while, but dying eventually from aspiration pneumonia as a result of inhalation of exogenous water. The cause of his death was, therefore, given as deferred death from drowning rather than aspiration pneumonia. PMID- 15335619 TI - The Policy Advisory Board for Forensic Pathology. PMID- 15335620 TI - Current perspectives on young offenders: aliens or alienated? PMID- 15335621 TI - Fitness to be interviewed and the appropriate adult scheme: a survey of police surgeons' attitudes. AB - Police surgeons are frequently asked by the police to assess whether suspects held in police custody are fit to be interviewed. However, there are no established medical criteria for making these assessments and very little has been published on how police surgeons actually approach the question of assessing a suspect's fitness for interview. PMID- 15335622 TI - Dutch and Scottish clinical forensic medicine: a comparison. AB - In the light of moves towards European union, we have greater opportunities to benefit from the experiences of our European colleagues and to formulate a 'best practice' policy as a result of the sharing of ideas. This paper compares the work of clinical forensic examiners in Scotland and in the Netherlands. It highlights the lessons learnt from observing Dutch practice and concludes by identifying the ones that could usefully be adopted in Scotland. PMID- 15335623 TI - Wound patterns: detection, documentation and analysis. AB - Bruises and other injuries located in skin can play a valuable role in the forensic investigation of a crime. This paper present details outlining how forensic odontology can be useful in detecting, documenting and analyzing such patterned injuries. In addition, four illustrated case histories are included. PMID- 15335624 TI - The role of a forensic clinical nurse specialist in a sexual assault treatment program. AB - The role of a Forensic Clinical Nurse Specialist has been developed within our Regional Sexual Assault Treatment Centre in London, Ontario, to assist in meeting the short-term and long-term physical and psychological needs of victims of acute sexual assault. We report on the development and application of this most valuable role within our medical and nursing health care team. PMID- 15335625 TI - Management of diabetes in clinical forensic practice. PMID- 15335626 TI - Assessment and management of individuals under the influence of alcohol in police custody. AB - Individuals taken into police custody are commonly under the influence of alcohol. Some of these individuals are dependent on alcohol and suffer from withdrawal symptoms. Morbidity and mortality in this group is high, and deaths have been recorded in police custody. There are no specific guidelines on the assessment and management of individuals under the influence of alcohol taken into police custody. This article outlines the clinical features, assessment, investigations and management of these individuals based on a review of the literature. PMID- 15335627 TI - Inhalation of steaming seafood aroma: sudden death in an asthmatic. AB - A 29-year-old asthmatic with a family history of atopy inhaled a large dose of fumes from sizzling, steaming seafood being served to another client in a restaurant. She immediately developed respiratory distress, loss of consciousness and cyanosis; she also vomited profusely, aspirated and died with 58 min of the onset of symptoms. Autopsy revealed epiglottitis and tracheobronchitis associated with an infiltration by mast cells. The blood IgE was marginally elevated. Death was attributed to upper respiratory airway obstruction due to aspiration of gastric contents, and to bronchospasm following the inhalation of steaming vapour from the seafood dish. The practice of transporting a platter of sizzling, steaming seafood across dining rooms and restaurants with its aroma permeating the entire premises could prove fatal for atopic customers. PMID- 15335628 TI - Drink-drive: broken jaw, lager and khat ingestion. AB - The report describes an examination of a man concerning an offence under The Road Traffic Act 1988. This was undertaken for two reasons, firstly to assess whether the suspect's alleged facial and jaw injuries prevented him from using the breathalyser and secondly to determine whether the man's ability to drive a motor vehicle was impaired. He admitted to drinking lager and chewing khat. The outcome in the magistrate's court is described. The use and effects of khat is reviewed. PMID- 15335629 TI - Suicide by proxy: a case report of juvenile autoerotic sexual asphyxia disguised as suicide. A common occurrence? AB - The confusion and embarrassment felt by a person discovering a body who has died through sexual asphyxia is likely to be considerable. Attempts may be made to disguise the nature of the death to medical attendants and to investigating police officers. A high level of suspicion and a knowledge of the characteristic findings seen in sexual asphyxia are important for the correct determination of the cause of death. This report describes the death of a 12-year-old boy in the Midlands, UK, in 1977. An open verdict had been recorded by the coroner. I Further evidence coming to light 17 years later appears to suggest that death was in fact due to autoerotic sexual asphyxia. PMID- 15335630 TI - The use of an alcometer in clinical forensic practice. AB - Forensic physicians are at times required to provide a professional opinion on a person's degree of intoxication through alcohol, particularly with regard to fitness for interview by the police. Inherent to these assessments is an estimation of the blood alcohol concentration (BAC). Most forensic physicians rely on taking a history regarding recent alcohol consumption and a clinical assessment to estimate the BAC. We report a study in which the BAC of 118 detainees was estimated and compared with the BAC measured by an alcometer. Our estimations were accurate in 66% of cases. Most errors resulted from underestimating the BAC. Additionally, we attempted to determine fitness for interview using a single measurement of the BAC without a clinical assessment. 19% of those who were assessed as unfit for interview had a BAC of less than the UK legal limit for driving (currently 80 mg%) and 37% considered fit, had a BAC above that level. We conclude that the routine use of alcometers would enhance the practice of forensic medicine but could not replace its role in the assessment of levels of alcohol intoxication. PMID- 15335631 TI - Compulsory random drug testing of prisoners in England and Wales: design flaws in the system. AB - The Home Office has recently introduced compulsory testing of prisoners in England and Wales for drug abuse. From 1996 all prisons in the UK will be involved. Urine samples from approximately 10% of the prison population will be collected each month. The method of drug analysis selected by the Home Office is fast and economical but readily prone to interference from common substances giving false results. An elaborate procedure has therefore been evolved including a rigorous personal search of the prisoner to prevent sample adulteration. The definitive test gas chromotography - mass spectrometry (GC-MS), is more expensive but is resistant to sample adulteration, and currently all positive samples are confirmed by this method. In view of the proportion of samples that have tested positive, the extent of the unknown number of false negatives, and the possible rejection of the collection protocol by prisoners, savings could be made if the method of analysis employed in the first instance was GC-MS. This paper illustrates the inaccuracies produced in an assay technique similar to that used by the Home Office when urine is contaminated by simple, commonly available substances. PMID- 15335632 TI - Fabricated homicides in Israel in light of the Intifada. AB - Fabrication of circumstances surrounding fatal events is not uncommon in forensic medicine. Reasons for this may include financial gain, shame and guilt. The use of a specific sociopolitical phenomenon as a fabricated scenario for lucrative purposes is reported. The importance of thorough scene of crime investigation and documentation, as well as a meticulous autopsy in order to determine the manner of death is emphasized. PMID- 15335633 TI - Fracture of the hyoid bone in survivors of attempted manual strangulation. AB - Fractures of the hyoid bone are known to occur in 17-71% of fatalities following manual strangulation, but are infrequently recognised in survivors. Two cases are presented of attempted manual strangulation in young women (aged 30 and 35 years) in which lateral neck X-rays disclosed an isolated fracture of the greater horn of the hyoid bone. In both cases the X-rays were performed because the victim complained of symptoms: one victim complained of pain on swallowing and on neck movement. The other complained of pain on speaking and swallowing. Radiological demonstration of an isolated hyoid fracture is of great evidential value in criminal proceedings in cases of attempted manual strangulation, especially when presentation to the police authorities is delayed and external evidence of neck injury may be minimal. Consideration should be given to obtaining X-rays in those cases where symptoms persist. PMID- 15335634 TI - Assessing cardiac risks in police officers. AB - It is necessary to assess the cardiovascular risks of police officers in order to address the issue of police officer safety, as well as the issue of public safety. In the past there has been a tendency to determine these risks according to the presence or absence of demonstrable ischaemia. Increased understanding of pathophysiology of coronary vascular disease leads to a different approach to this problem. This modified approach is discussed. PMID- 15335635 TI - Determining the true cause of death in a dermatological disaster. AB - Adverse drug reactions that result in patient death warrant special consideration to determine if the outcome was preventable. We report the results of an investigation into the cause of death of a 63 year old male who was thought to have phenytoin-induced toxic epidermal necrolysis (TEN). Most dermatological reactions from drugs are minor and resolve without sequelae once the drug is discontinued. In some cases, reactions are severe and can be life threatening. The patient arrived at the emergency department with extensive exfolative dermatitis. The differential diagnosis included phenytoin-induced TEN, scalded skin syndrome, and phenytoin hypersensitivity. After he was admitted his clinical status deteriorated and he died 13 days after admission. Autopsy findings were significant for necrotizing dermatitis, necrotizing pneumonia, multiple herpetic ulcerations, multisystem organ failure and blood cultures grew Staphylococcus aureus (TSS toxin positive). Findings that indicate a cause of death other than a drug-induced dermatological reaction are presented as well as an overview of the patients' medical history. The differential diagnosis of drug induced skin lesions are also discussed. PMID- 15335636 TI - Fatal firearm injuries caused by handmade weapons. AB - Six cases of accidental death caused by a shot from handmade or improvised firearms are described. Data were collected on incident, victim and special autopsy findings. In three cases the weapon used was a so-called 'pen gun' of small calibre (5.6 mm) and in two cases a home-made shotgun/muzzle loader with shotgun cartridge (containing 16 mm pellets) was used. In one case, the weapon was a shop-made gun with home-made ammunition. It is perhaps surprising that in the present war situation in Croatia, when all kinds of firearms are easily available, people continue to produce handmade weapons. PMID- 15335638 TI - Guidelines on the use of photographs and other materials obtained during pathological examinations. PMID- 15335637 TI - Defenestration and dehiscence: suicide or homicide? AB - Wound rupture or dehiscence is usually encountered clinically following failure of healing after surgery or less often after major trauma. This article describes a case of wound dehiscence resulting from a fall from a height. In this incident there was initial uncertainty concerning the manner of death. In view of a gaping abdominal wound, the police officers first on the scene were suspicious that the deceased had been the victim of an assault. PMID- 15335639 TI - The short report on 'Salivary alcohol measurement: use in clinical forensic medical practice'. PMID- 15335641 TI - Sudden infant death syndrome - a 'diagnosis' in search of a disease. AB - Although Sudden Infant Death Syndrome (SIDS) is the major cause of death in infants aged between 1 week and 1 year in Western countries, it is one of the most enigmatic conditions encountered in paediatric forensic practice. SIDS has been recognized since Biblical times, and yet the definition continues to be debated and the aetiology remains obscure. In addition, there are no accepted pathognomonic features at post-mortem and the diagnosis is still one of exclusion. Emery once asked whether the term 'SIDS' is in reality more of a 'diagnostic dustbin' into which are placed a variety of unrelated entities.(1) To a certain extent this is true, as it is now recognized that a range of disorders can result in the sudden and unexpected death of an infant in a cot. It is also likely that the aetiology of SIDS is heterogeneous and that the term SIDS is not so much a diagnosis but a term covering a variety of mechanisms which result in a common lethal outcome. There are a number of controversies in the SIDS field which complicate the use of the term and which confound the assessment of causes and mechanisms of sudden infant death. PMID- 15335642 TI - Excoriations and contusions of the skin as artefacts in fictitious sexual offences. AB - Some women reporting a sexual offence to the police inflict injuries on themselves, usually with the help of pointed and/or cutting tools, in order to make fictitious assaults more credible. If there is a classical pattern of the findings, such fictitious assaults are easier to identify. However, it is more difficult to distinguish genuine from fictitious injuries, if blunt force is used. This paper reports on 4 such cases, and the characteristics of self inflicted excoriations and skin contusions are discussed. In the cases described, the excoriations were produced by scratching with the person's own fingernails or by rubbing the skin against a rough surface; contusion bleeding was caused by pinching (lifting and compressing skin folds with the fingers). The pattern of the injuries showed striking parallels to classical patterns: multiple lesions in grouped, parallel and symmetrical arrangement and of evenly minor intensity; preference of body regions within easy reach; no damage of the clothing, or if any, damage inconsistent with the injuries as described. PMID- 15335643 TI - Computer simulation analysis of blood alcohol. AB - A mathematical model using an iterative algorithm based on the Widmark formula is used to simulate a continuous spectrum of the expected blood alcohol concentration from start of alcohol dosage to time of specific event in numerical and graphic form. Absorption is based on flexible first order rates with provision for an optional good-fit model of the delaying effect of food. Elimination is based initially on selectable zero order rates followed by first order kinetics at low blood alcohol concentrations. A correlation coefficient of 0.94 was obtained in comparisons of projected to observed blood alcohol concentrations. A measurement of the area under the curve is provided. The software program utilizing the algorithm is suitable for forensic and clinical applications. PMID- 15335644 TI - Deaths from methadone use. AB - All deaths from methadone poisoning occurring between 1991 and 1994 in Sheffield, UK, were studied. There were 18 deaths: 17 adults (14 men, 3 women), age range of 17-45 years (mean 29), and a 3-year-old female child. Blood methadone concentrations ranged from 200 microg/l to 1863 microg/l (mean 560 microg/l). Multiple drug use was common. Ten people had been prescribed methadone, whilst eight had obtained their methadone illegally. Only 3 people were long-term methadone users. The others were probably first time users or recently new to methadone programmes. Two, including the child, were apparently forced to drink methadone. Methadone is a potent drug with serious side-effects. Naive users are particularly at risk of death. Toxic effects may be delayed, allowing large amounts to be ingested, giving both user and medical practitioner a false sense of security. Caution should be exercised when prescribing to new patients in methadone programmes. Controlled administration of methadone may prevent overdosage and ingestion by those to whom it is not prescribed. PMID- 15335645 TI - Forensic dentistry in the identification of victims and assailants. AB - The teeth are the least destructible part of the body and may remain more or less intact for many years beyond death. In addition, because of their stability in a biological sense, and because during their formation disease may affect the hard tissues of the teeth themselves, they contain information about the physiological and pathological events in the life of the individual which remain as markers within the hard tissues of the teeth. Therapeutic activity by a dentist in the form of restorations and prostheses may modify an individual's dentition in a more or less unique manner. It is the role of the forensic dentist to assess this biological and chemical information, and use it in the identification of an unknown body. The teeth may also be used to inflict injury on a victim in the form of a bite. The resulting bite mark may not only indicate the degree of aggression during the assault but if sufficient details are available within it, may enable the forensic dentist to determine the identity of the assailant. This important aspect of forensic dentistry is commonly required in non-accidental injuries to children and in instances of grievous bodily harm, rape and murder of child or adult. PMID- 15335646 TI - Penetrating spinal injury inflicted by screwdriver: unusual morphological findings. AB - A 50-year-old female victim died of recurrent pulmonary embolism, 3 months after having received multiple screwdriver stab wounds in the neck and upper spine. Autopsy showed that one stab had penetrated the middle part of the cervical spinal cord and thus caused an incomplete tetraplegia. In the region of the healed spinal cord and the affected meninges, a considerable amount of hair and textile fibres surrounded by foreign body giant cells and elastic fibres of spinal ligament were found, all of which had been carried into the stab canal by the blunt tip of the screwdriver. In addition to the primary clinical findings, this unusual transportation of matter into the depth of the stab canal allowed identification of the murder weapon. PMID- 15335647 TI - Physical ability, fitness and police work. AB - The objective of this article is to provide an overview of physical ability, fitness and police work. The literature will be reviewed, and the method we have used to develop a tool that measures the physical abilities required for police work will be presented. The importance of linking this standard with a programme for health promotion will be stressed. The reasons why this standard is occupation specific and non-discriminatory will be explained. PMID- 15335648 TI - Ethical duty to report abuse of prisoners. PMID- 15335649 TI - Introduction to the Fifth Cross Channel Conference needs to be rectified. PMID- 15335650 TI - The use of force, chemical agents and restraint in a prison psychiatric setting. AB - An attempt is made to delineate some parameters surrounding the use of force within a psychiatric service to a correctional system. Descriptive study in using case material and incident statistics are followed by discussion. Reference is made to some recent articles on ethics in the medical and psychiatric literature. This article is offered in recognition that examining these troublesome issues can be very difficult for the clinicians involved, and not in the expectation that any ready solutions are to be found. PMID- 15335651 TI - Screening of volatile substances and determination of toluene (a thinner component) in the blood and urine in emergency medical care and autopsy cases by the pulse heating method. AB - Screening of volatile substances was performed by pulse heating gas chromatography - mass spectrometry (GC-MS) using a GS-Q column in 211 emergency medical care and 342 autopsy cases. At least 36 standard substances could be separately detected. Six kinds of volatile substances were screened in a total of 553 cases. Toluene and/or hippuric acid were detected in the blood and/or urine in respectively, 4 emergency medical care and 8 autopsy cases. There were 11 abusers (9 males and 2 females) in these 12 positive cases. The ages of the abusers ranged from 13-26 years. There was no particular pattern to the monthly frequency distribution of identification of thinner (toluene) abuse cases, which occurred throughout the year. It is believed that these data at least partly reflect the present status of thinner/glue abuse in Japan. We conclude that pulse heating GC-MS is useful in the screening and quantitative determination of volatile substances including toluene and other thinner/glue components. PMID- 15335652 TI - The role of an operating microscope in medico-legal practice. AB - This article describes our experience in the use of a binocular surgical microscope at autopsy for the in situ forensic examination of various types of mechanical skin wounds. The microscopic image was visualised on a closed-circuit television monitor, and colour photographs and video recordings were simultaneously obtained from the monitor. With abrasions, small epidermal tags, which play a key role in the determination of external force direction, were clearly observed. With lacerations, crushing of the epidermis was observed at the wound margin, and continuous bridges of connective tissue and/or blood vessels were also found within the wound cavity. With incised wounds, the epidermis was intact up to the wound margin, and there was no continuous bridge within the wound cavity. Using these differences, it was, therefore, simple to distinguish lacerations from incised wounds. Ligature marks due to hanging were clearly observed, with petechial haemorrhages and minute abrasions readily seen in the lesion. For stab wounds caused by a single-edge knife, the morphological difference between the two ends of the wound caused by the sharp blade and the blunt side could be distinguished. It was also possible to confirm the identification of unknown cadavers by observing old wound scars using the operating microscope. Abnormal lesions of the blood vessels, such as a mechanical tear of the vertebral artery (the origin of a subarachnoid haemorrhage) and the atherosclerotic stenosis of the coronary artery, were detectable. In addition, the microscope was useful as a teaching aid to colleagues and medical students to study the nature of the wounds during or after the autopsy. We believe that this system presents advantages in terms of education and training of forensic pathologists and physicians as well as crime investigation officers. PMID- 15335653 TI - The role of a community psychiatric nurse in prison healthcare. AB - It is now 2 years since a trained and experienced psychiatric nurse became attached to our Health Care Centre to assist in counselling and treating mentally disordered offenders. The attachment represents innovation within the Prison Service and the nurse has developed into a most valuable asset within the health care team. We report on the attachment, the workload and the role of the nurse. PMID- 15335654 TI - Forensic physicians and medical confidentiality: a brief summary. PMID- 15335655 TI - Perinatal death as a consequence of fetal stabbing: was it murder? AB - At 22 weeks gestation, a fetus received a stab wound to the abdomen when the mother was subjected to a knife attack. Premature labour followed 2.5 weeks later. The baby received full intensive care, but died at 4.5 months of age from complications directly related to the premature birth. The person who committed the assault was indicted with murder of the infant according to the precept of transferred malice. In the absence of any clear legal precedent, the judge ruled that the indictment was not consistent with the principles of criminal law, since the malice directed towards the mother could not be regarded as being transferred to the fetus. This was because the wounding of a non-viable fetus with death following 4 months later did not amount to a deliberate killing, and in addition, it was not the intention of the Defendant at the time of the stabbing to kill the infant. PMID- 15335656 TI - Taking samples under the Police and Criminal Evidence Act 1984. AB - This article reviews the changes that have been made in England and Wales by Part IV of the Criminal Justice and Public Order Act 1994 to the powers conferred on the police by the Police and Criminal Evidence Act 1984 to take and to retain for statistical purposes intimate and non-intimate samples from suspects. These changes follow recommendations made by the Royal Commission on Criminal Justice which reported in 1993. They signal a significant increase both in police powers and in the future work of the police surgeon. PMID- 15335658 TI - Occupational medicine for policing. AB - A wide variety of medical conditions may be associated with police work. Unlike other occupations where a specific link can be traced between an exposure or an action and a specific pathology, the link between police work and disease is more problematic. The medical conditions which seem to be associated with police work are all conditions for which numerous other risk factors are identifiable. These risk factors include physical inactivity, poor nutritional practices, cigarette smoking and alcohol overuse. While it is undoubtedly desirable to minimise these risk factors in any patient population it is imperative, given the increased risks among police personnel, that aggressive attempts he undertaken to reduce cumulative risks. The physician plays an important part in this process by screening for specific conditions associated with police work, by educating the police officer about increased risks and by encouraging lifestyle choices that will reduce risk. It is also important that the physician encourages the use of personal protective equipment where appropriate. Periodic health assessment of police officers by a physician knowledgeable about police work should include education about the risks associated with the occupation and about methods to reduce risk. The physician should also enquire about exposures to violent or dangerous occurrences and should include assessment for possible emotional sequelae of such exposure. Operational procedures designed to reduce risk of violence and to improve police officer safety and survival are appropriate but are not in the normal realm of the physician. The question of whether a medical condition may be attributable to the occupation of policing is liable to produce strong emotions. When a police officer becomes ill there is an understandable desire on the part of other officers and, often, on the part of the public, to attempt to demonstrate a connection between the occupation and the illness. In line with this tendency several US states have policies in place to unquestioningly accept atherosclerotic heart disease among police officers as 'occupationally induced' for pension purposes. This leads to situations where the scientific evidence may be at odds with the political agenda of individuals and groups participating in the determination. This review will examine the evidence for a number of medical conditions. Specific occupational exposures, such as lead exposure among ballistics specialists, or chemical exposure among forensic laboratory workers are addressed elsewhere. Biohazard risks will be addressed in a later review. PMID- 15335659 TI - Risk to police officers from biohazards encountered in police work. AB - The risk of contracting infectious disease in the course of police work is reviewed. The specific focus is on Acquired Immune Deficiency Syndrome (AIDS), hepatitis and tuberculosis. A review of the relevant literature and an assessment of the biohazard risk specific to police work is provided. The risk of AIDS and hepatitis C is seen to be less than the risk of hepatitis B. For these blood borne diseases, universal precautions are recommended. The application of such precautions to policing is outlined. Immunisation against hepatitis B is recommended. Tuberculosis is seen as a possibly increasing risk. A programme of surveillance is recommended. A review of post-exposure management is provided. PMID- 15335660 TI - Elder abuse in the United States. AB - Elder abuse in the US became a public issue in 1978, due to investigation by the US House of Representatives Subcommittee on Family Violence. Defining abuse has proved difficult, but five primary categories have been developed: physical, psychological, financial/material abuse, violation of personal rights, and neglect. Abuse of some type occurs in 4-10% of Americans over 65 years of age. There are presently laws in all 50 states dealing with elder mistreatment. Mandatory reporting has been enacted in most states; however, most laws provide no funds for meaningful intervention. Recommendations for preventing, diagnosing, and reporting elder mistreatment are reviewed. Intervention requires a multidisciplinary effort by professionals from the medical, social service, mental health and legal professions. PMID- 15335662 TI - Clinical forensic medical services: London and Melbourne contrasted. AB - A personal comparison and contrast of the differences between two geographical areas following a unique opportunity to experience working in clinical forensic medicine in both London and Melbourne. PMID- 15335661 TI - Clozapine treatment of schizophrenia: implications for forensic psychiatry. AB - Clozapine, a new antipsychotic medication, is now the first-line treatment for neuroleptic refractory schizophrenia. Preliminary observations on its efficacy in treating schizophrenic patients with co-morbid substance abuse and, particularly, schizophrenic patients with persistent aggressive behaviour suggests that clozapine may provide a useful treatment option in forensic patient populations. Potential applications of clozapine and management considerations that are relevant to the forensic setting are discussed. PMID- 15335663 TI - Screening and determination of methamphetamine and amphetamine in the blood, urine and stomach contents in emergency medical care and autopsy cases. AB - Methamphetamine (MA) and amphetamine (AMP) were screened and their levels were determined using the Toxi-Lab thin-layer chromatography system and gas chromatography-mass spectrometry, respectively, in the blood, urine and stomach contents from 211 emergency medical care and 417 autopsy cases. MA and AMP were detected in 5 emergency medical cases, and the blood MA and AMP concentrations ranged from 0.697-0.041 micromol/100 g and from 0.0944-0.0003 micromol/100 g, respectively. MA and AMP were detected in 19 autopsy cases, in which blood MA and AMP concentration ranged from 14.3-0.123 micromol/100 g and from 0.256-0.0017 micromol/100 g, respectively. The autopsy cases included 5 cases of sudden death with blood MA concentration of less than 3 micromol/100 g. MA and AMP screening and determination in emergency medical care and autopsy cases provide useful information and are indispenable in clarifying the dimensions of MA abuse in Japan. PMID- 15335664 TI - Impact of antiseizure medication level on the incidence of sudden unexplained death from epilepsy in patients from an inner city emergency department. AB - Sudden unexplained death from seizures (SUDS) accounts for death in approximately 10% of the epileptic population. SUDS usually occurs in young males with a history of seizure disorders who are in otherwise good health. No definitive anatomical lesions are found at autopsy that explain death. There is however, a correlation between SUDS and subtherapeutic levels of antiseizure medications. The purpose of this study was to retrospectively review and compare drug levels from a seizure patient that presented to an inner city emergency department to those from the medical examiners office. This study was prompted by a wrongful death claim for substandard care in a known seizure disorder patient. The claim alleged that the death was directly attributed to subtherapeutic seizure medication levels. We report the results from 150 seizure patients that presented to the emergency department with a chief complaint of a recent seizure and of 163 patients that were examined post mortem. 58% of the emergency department patients who were taking phenytoin, 79% taking carbamazapine and 82% of the patients taking phenobarbital had subtherapeutic levels. No patient in this population died and only 13% required hospital admission. These levels were comparable to the post mortem population. PMID- 15335665 TI - Salivary alcohol measurement: use in clinical forensic medical practice. AB - This report describes the use of a salivary alcohol measurement device in clinical forensic medicine, and concludes that it may be of use in assisting the diagnosis or the assessment of patients in the custodial setting. PMID- 15335666 TI - Acupuncture and its therapeutic use with criminal justice populations. PMID- 15335667 TI - The European council of legal medicine. PMID- 15335668 TI - Ethical duty to report abuse. PMID- 15335669 TI - Ecological separation and genetic isolation of Neisseria gonorrhoeae and Neisseria meningitidis. AB - BACKGROUND: Classifying bacteria into species is problematic. Most microbiologists consider species to be groups of isolates that share some arbitrary degree of relatedness of biochemical or molecular (such as DNA sequence) features and that, ideally, are clearly delineated from all other groups of isolates. The main problem in applying to bacteria a biological concept of species based on the ability or inability of their genes to recombine, is that recombination appears to be rare in bacteria in nature, as indicated by the strong linkage disequilibrium between alleles found in most bacterial populations. However, there are some naturally transformable bacteria in which assortative recombination appears to be so frequent that alleles are in, or close to, linkage equilibrium. For these recombining populations a biological concept of species might be applicable. RESULTS: Populations of Neisseria gonorrhoeae and Neisseria meningitidis from Spain were analysed by multilocus enzyme electrophoresis. The data indicate that assortative recombination occurs frequently within populations, but not between populations. Similarly, the sequences of two house-keeping genes show no evidence of intragenic recombination between N. gonorrhoeae and N. meningitidis. CONCLUSIONS: N. gonorrhoeae and N. meningitidis represent extremely closely related 'sexual' populations that appear to be genetically isolated in nature, and thus conform to the biological concept of species. The extreme uniformity of N. gonorrhoeae house-keeping genes suggests that this species may have arisen recently as a clone of N. meningitidis that could colonize the genital tract. Ecological isolation - of populations that can colonize the genital tract from those that can colonize the nasopharynx - may have been an important component in speciation, leading to a lower frequency of recombination between species than within species. PMID- 15335670 TI - Hematopoietin sub-family classification based on size, gene organization and sequence homology. AB - The hematopoietins constitute a family of structurally related proteins that includes many cytokines, growth factors and hormones. We propose a classification for hematopoietins based on their size, gene organization and sequences; in addition, we further suggest that three cytokines, IL-11, IL-12 alpha chain and IL-13, are hematopoietins, and we place them in specific subgroups. PMID- 15335671 TI - Dorso-ventral patterning in the neural tube. PMID- 15335672 TI - Seeing double. PMID- 15335673 TI - The structure of a molecular motor. PMID- 15335674 TI - Righting reading. PMID- 15335675 TI - Integrins as virus receptors. PMID- 15335676 TI - Do trade-offs hold the key? PMID- 15335677 TI - Emerging families of cytokines and receptors. PMID- 15335678 TI - Knock-out but not knocked out. PMID- 15335679 TI - A new DNA quadruplex. PMID- 15335681 TI - Hox genes and the hindbrain. PMID- 15335680 TI - Excitement over inhibition. PMID- 15335682 TI - A kinase that responds to stress. PMID- 15335683 TI - The virtue of complexity. PMID- 15335684 TI - What is evolution playing at? PMID- 15335685 TI - When everyone's map is different. PMID- 15335686 TI - Does oligodendrocyte survival depend on axons? AB - BACKGROUND: We have shown previously that oligodendrocytes and their precursors require signals from other cells in order to survive in culture. In addition, we have shown that about 50% of the oligodendrocytes produced in the developing rat optic nerve normally die, apparently in a competition for the limiting amounts of survival factors. We have hypothesized that axons may control the levels of such oligodendrocyte survival factors and that the competition-dependent death of oligodendrocytes serves to match their numbers to the number of axons that they myelinate. Here we test one prediction of this hypothesis - that the survival of developing oligodendrocytes depends on axons. RESULTS: We show that oligodendrocyte death occurs selectively in transected nerves in which the axons degenerate. This cell death is prevented by the delivery of exogenous ciliary neurotrophic factor (CNTF) or insulin-like growth factor I (IGF-1), both of which have been shown to promote oligodendrocyte survival in vitro. We also show that purified neurons promote the survival of purified oligodendrocytes in vitro. CONCLUSION: These results strongly suggest that oligodendrocyte survival depends upon the presence of axons; they also support the hypothesis that a competition for axon-dependent survival signals normally helps adjust the number of oligodendrocytes to the number of axons that require myelination. The identities of these signals remain to be determined. PMID- 15335687 TI - One-step site-directed mutagenesis of the Kex2 protease oxyanion hole. AB - BACKGROUND: Members of the subtilisin family of serine proteases usually have a conserved asparagine residue that stabilizes the oxyanion transition state of peptide-bond hydrolysis. Yeast Kex2 protease is a member of the subtilisin family that differs from the degradative subtilisin proteases in its high substrate specificity, it processes pro-alpha-factor, the precursor of the alpha-factor mating pheromone of yeast, and also removes the pro-peptide from its own precursor by an intramolecular cleavage reaction. Curiously, the mammalian protease PC2, a Kex2 homolog that is likely to be required for pro-insulin processing, has an aspartate in place of asparagine at the 'oxyanion hole'. RESULTS: We have tested the effect of making substitutions of the conserved oxyanion-hole asparagine (Asn 314) of the Kex2 protease. To do this, we have developed a rapid method of site-directed mutagenesis, involving homologous recombination of a polymerase chain reaction product in yeast. Using this method, we have substituted alanine or aspartate for Asn 314 in a form of Kex2 engineered for secretion. Transformants expressing the two mutant enzymes could be identified by failure either to produce mature alpha-factor or to mate. The Ala 314 enzyme was unstable but the Asp 314 enzyme accumulated to a high level, so that it could be purified and its activity towards various substrates tested in vitro. We found that, with three peptides that are good substrates of wild-type Kex2, the k(cal) of the Asp 314 enzyme was reduced approximately 4500-fold and its K(M) approximately 4-fold, relative to the wild-type enzyme. For the peptide substrate corresponding to the cleavage site of pro-alpha-factor, however, k(cat) of the Asp 314 enzyme was reduced only 125-fold, while the K(m) was increased 3 fold. Despite its reduced catalytic activity, however, processing of the mutant enzyme in vivo - by the intramolecular cleavage that removes its amino-terminal pro-domain - occurs at an unchanged rate. CONCLUSIONS: The effects of the Asn 314 Asp substitution reveal contributions to the reaction specificity of the Kex2 protease of substrate residues amino-terminal to the pair of basic residues at the cleavage site. Aspartate at the oxyanion hole appears to confer k(caf) discrimination between substrates by raising the energy barrier for productive substrate binding: this may have implications for pro-insulin processing by the PC2 protease, which has an aspartate at the equivalent position. The rate of intramolecular cleavage of pro-Kex2 may be limited by a step other than catalysis, presumably protein folding. PMID- 15335689 TI - How many neurons does it take to see? PMID- 15335688 TI - Agrobacteria mate in opine dens. PMID- 15335690 TI - Networking with protein kinases. PMID- 15335691 TI - Fidelity in mRNA splicing. PMID- 15335692 TI - More to colour than meets the eye. PMID- 15335693 TI - Mapping the brain with MSI. PMID- 15335694 TI - Not in the thymus. PMID- 15335695 TI - Flexible friends. PMID- 15335696 TI - Dodging the genes. PMID- 15335697 TI - Clues from clueless. PMID- 15335698 TI - Will the real IP4 receptor please stand up? PMID- 15335699 TI - Clear as crystal? PMID- 15335700 TI - Understanding erythroid differentiation. PMID- 15335701 TI - Reworking an old brain. PMID- 15335702 TI - Transcriptional complexity. PMID- 15335703 TI - Stimulating the lymphocytes. PMID- 15335704 TI - Second generation enhancer traps. PMID- 15335705 TI - Verbal and visual subsystems of working memory. PMID- 15335706 TI - The viral canyon. PMID- 15335707 TI - Generation of normal lymphocyte populations by Rb-deficient embryonic stem cells. AB - BACKGROUND: Mice homozygous for a loss-of-function mutation of the recombination activating gene-2 (RAG 2), which is required for the rearrangement of antigen receptor genes, do not produce mature B and T lymphocytes. But chimeric mice that result from injection of normal embryonic stem (ES) cells into blastocysts from RAG2-deficient mice develop normal mature lymphocyte populations, all of which are derived from the injected ES cells; we have called this process RAG2 deficient blastocyst complementation. Using ES cells with homozygous mutations, RAG-2-deficient blastocyst complementation could provide a physiological assay with which to determine the potential role of almost any gene in the development and/or function of lymphocytes. To test the general utility of this system, we have used it to test the differentiation-potential of ES cells that harbor homozygous loss-of function mutations of their retinoblastoma susceptibility (Rb) gene loci. We chose Rb for this analysis because of its widespread function in the control of the cell cycle and cell differentiation, the adverse effect of homozygous germline mutations of Rb on hematopoiesis in fetal liver, and the embryonic lethality that results when the homozygous Rb mutation is introduced into the germline. RESULTS: Homozygous Rb mutant ES cells can develop into phenotypically normal, mature B and T lymphocytes in the RAG-2-deficient background. Strikingly, Rb-deficient B and T cells do not have major defects in either activation or function. CONCLUSION: We have demonstrated the efficacy of the RAG-2-deficient blastocyst complementation system for evaluating the role of critical genes in lymphocyte development. Our results indicate that Rb expression is not intrinsically required for B-cell or T-cell function, despite the normally high levels of Rb expressed in lymphoid cells. PMID- 15335708 TI - A method for predicting protein structure from sequence. AB - BACKGROUND: The ability to predict the native conformation of a globular protein from its amino-acid sequence is an important unsolved problem of molecular biology. We have previously reported a method in which reduced representations of proteins are folded on a lattice by Monte Carlo simulation, using statistically derived potentials. When applied to sequences designed to fold into four-helix bundles, this method generated predicted conformations closely resembling the real ones. RESULTS: We now report a hierarchical approach to protein-structure prediction, in which two cycles of the above-mentioned lattice method (the second on a finer lattice) are followed by a full-atom molecular dynamics simulation. The end product of the simulations is thus a full-atom representation of the predicted structure. The application of this procedure to the 60 residue, B domain of staphylococcal protein A predicts a three-helix bundle with a backbone root mean square (rms) deviation of 2.25-3 A from the experimentally determined structure. Further application to a designed, 120 residue monomeric protein, mROP, based on the dimeric ROP protein of Escherichia coli, predicts a left turning, four-helix bundle native state. Although the ultimate assessment of the quality of this prediction awaits the experimental determination of the mROP structure, a comparison of this structure with the set of equivalent residues in the ROP dime- crystal structure indicates that they have a rms deviation of approximately 3.6-4.2 A. CONCLUSION: Thus, for a set of helical proteins that have simple native topologies, the native folds of the proteins can be predicted with reasonable accuracy from their sequences alone. Our approach suggest a direction for future work addressing the protein-folding problem. PMID- 15335710 TI - SH2 and SH3 domains. PMID- 15335709 TI - Simple and efficient generation of marked clones in Drosophila. AB - BACKGROUND: Cell lineage analysis and mosaic analysis of mutations are important techniques that are used to study the development of many organisms. Unfortunately, the methods employed for such analyses are usually inefficient, technically demanding or labor intensive. In Drosophila, the most common methodology used for the generation of mosaic animals is mitotic recombination, which is induced by X-rays. Although this technique is simple, it has the undesirable characteristics of a low efficiency and a high rate of cell death. Furthermore, although a large number of marker systems has been employed to detect mitotic recombinants, none allows easy identification of clones for all cell types. RESULTS: A system is described here that allows a highly efficient generation of clones with the concomitant expression of an easily detectable cellular marker. This method can be applied to cell lineage and mosaic analysis in Drosophila. The site-specific yeast FLP recombinase, under the control of a heat shock-inducible promoter, efficiently catalyses mitotic recombination specifically at the site of a FLP recombination target (FRT). In this system, recombination fuses the alpha-tubulin promoter to the lacZ gene, allowing transcription of the marker. Recombinant cells and their progeny can, therefore, be detected by standard assays for beta-galactosidase. Of particular importance is the fact that only the cells of interest stain, thus allowing their simple detection in any tissue. CONCLUSIONS: We demonstrate that, by intermolecular recombination, we can use FLIP recombinase to generate marked clones efficiently in embryonic, larval and adult tissues. This simple and efficient technique is well suited to cell-lineage analysis and can be easily extended to the generation and detection of mutant clones in mosaic animals. PMID- 15335711 TI - Patterning the Arabidopsis embryo. PMID- 15335712 TI - From outer ear to virtual space. PMID- 15335713 TI - A kinase with Ku-dos. PMID- 15335714 TI - Wizardry at the cell cortex. PMID- 15335715 TI - An expanding enigma. PMID- 15335716 TI - A second chance for bad B cells. PMID- 15335717 TI - Stroke recovery. PMID- 15335718 TI - More mutations in males? PMID- 15335719 TI - New primers for an old enzyme. PMID- 15335720 TI - Fixed or fluid? PMID- 15335721 TI - Much ado about docking. PMID- 15335723 TI - A tale of two toxins. PMID- 15335722 TI - Ret oncogene responsible for MEN2A. PMID- 15335724 TI - Constrained V gene choice. PMID- 15335725 TI - Rapid identification of proteins by peptide-mass fingerprinting. AB - BACKGROUND: Developments in 'soft' ionisation techniques have revolutionized mass spectro-metric approaches for the analysis of protein structure. For more than a decade, such techniques have been used, in conjuction with digestion b specific proteases, to produce accurate peptide molecular weight 'fingerprints' of proteins. These fingerprints have commonly been used to screen known proteins, in order to detect errors of translation, to characterize post-translational modifications and to assign diulphide bonds. However, the extent to which peptide mass information can be used alone to identify unknown sample proteins, independent of other analytical methods such as protein sequence analysis, has remained largely unexplored. RESULTS: We report here on the development of the molecular weight search (MOWSE) peptide-mass database at the SERC Daresbury Laboratory. Practical experience has shown that sample proteins can be uniquely identified from a few as three or four experimentally determined peptide masses when these are screened against a fragment database that is derived from over 50 000 proteins. Experimental errors of a few Daltons are tolerated by the scoring algorithms, thus permitting the use of inexpensive time-of-flight mass spectrometers. As with other types of physical data, such as amino-acid composition or linear sequence, peptide masses provide a set of determinants that are sufficiently discriminating to identify or match unknown sample proteins. CONCLUSION: Peptide-mass fingerprints can prove as discriminating as linear peptide sequences, but can be obtained in a fraction of the time using less protein. In many cases, this allows for a rapid identification of a sample protein before committing it to protein sequence analysis. Fragment masses also provide information, at the protein level, that is complementary to the information provided by large-scale DNA sequencing or mapping projects. PMID- 15335726 TI - Molecular mimicry of the antigen receptor signalling motif by transmembrane proteins of the Epstein-Barr virus and the bovine leukaemia virus. AB - BACKGROUND: Many transmembrane proteins of eukaryotic cells have only a short cytoplasmic tail of 10 - 100 amino acids, which has no obvious catalytic function. These tails are thought to be involved either in signal transduction or in the association of transmembrane proteins with the cytoskeleton. We have previously identified, in the cytoplasmic tails of components of B and T lymphocyte antigen receptors, an amino-acid motif that is required for signalling. The same motif is also found in the cytoplasmic tails of two viral proteins: the latent membrane protein, LMP2A, of Epstein Barr virus and the envelope protein, gp30, of bovine leukaemia virus. Interestingly, both viruses can activate infected B lymphocytes to proliferate, as does signalling by the B cell receptor. RESULTS: In this study, we show that the cytoplasmic tails of the two viral proteins, and the cytoplasmic tail of the B-cell receptor immunoglobulin-alpha chain, when linked to CD8 in chimeric transmembrane proteins, can transduce signals in B cells. Cross-linking of these chimeric receptors activates B-cell protein tyrosine kinases and results in calcium mobilization. Furthermore, these cytoplasmic sequences are also protein tyrosine kinase substrates and may interact with cytosolic proteins carrying SH2 protein protein interaction domains. CONCLUSION: Our findings suggest that viral transmembrane proteins can mimic the antigen-induced stimulation of the B-cell antigen receptor and thus can influence the activation and/or survival of infected B lymphocytes. PMID- 15335727 TI - The consequences of demographic reduction and genetic depletion in the endangered Florida panther. AB - The Florida panther has recently suffered severe range and demographic contraction, leaving a remarkably low level of genetic diversity. This exerts a severe fitness cost, manifested by spermatozoal defects, cryptorchidism, cardiac abnormalities and infectious diseases that threaten the survival of the subspecies. PMID- 15335728 TI - Gene regulation: coming to grips with cactus. PMID- 15335729 TI - Plant cell expansion: loosening the ties. PMID- 15335730 TI - Homologous recombination: a universal recombination filament. PMID- 15335732 TI - Zebrafish development: fluorescent markers shed light on cell lineage. PMID- 15335731 TI - Neural development: new ligands for Neu? PMID- 15335733 TI - Chemokines: combinatorial mediators of inflammation? PMID- 15335734 TI - Plant hormones: what a gas! PMID- 15335735 TI - Gene organization: nested genes take flight. PMID- 15335736 TI - RNA ligands: out of shape but fir for recognition. PMID- 15335737 TI - Touch: the code for roughness. PMID- 15335738 TI - Antigen presentation: untapped peptides. PMID- 15335739 TI - Cytosine methylation: the pros and cons of DNA methylation. PMID- 15335740 TI - Cytoskeleton: watching the tube. PMID- 15335741 TI - Apoptosis: the cell cycle and cell death. PMID- 15335742 TI - Organelle communication: intracellular dialogue. PMID- 15335743 TI - Visual perception: solid shape and the natural world. PMID- 15335744 TI - Polar zippers. AB - BACKGROUND: Certain proteins are known to form leucine zippers - alpha-helical coiled-coils in which the non-polar side chains of two leucine-rich helices intermesh. We recently presented the first evidence for a polar zipper, formed by the carboxy-terminal peptides of the eight subunits of Ascaris haemoglobin. The evidence was based on the presence of pairs of acidic residues alternating with pairs of basic residues ( + + - - ) in an amino-acid sequence that has since been shown to be incomplete. The complete sequence, derived from the haemoglobin's cDNA, now shows a self-complementary polar sequence extending along the entire length of its 24-residue carboxy-terminal peptide. RESULTS: From the complete sequence, it is clear that the eight identical subunits of the haemoglobin could be held together by an eight-stranded antiparallel beta barrel made up of the carboxy-terminal 24 residues of each of the subunits, such that each strand forms 10 salt bridges with each of its neighbours. A computer search of the protein database revealed similar, but shorter, + + - - repeats in several other proteins. It also revealed long repeats of alternating arginine and aspartate residues, and long stretches of only glutamines, or only serines, suggestive of several other kinds of polar zippers. CONCLUSION: Several proteins have amino acid sequences that suggest the formation of polar zippers made of beta strands. These could form antiparallel pleated sheets linked together by hydrogen bonds between polar side chains both above and below the plane of the sheets. Polar zippers may be important in welding together oligomeric proteins which have subunits lacking the extensive complementary surfaces necessary for stability, or in promoting the association of functionally complementary proteins. PMID- 15335745 TI - The origin of chromosome rearrangements at early stages of AMPD2 gene amplification in Chinese hamster cells. AB - BACKGROUND: Gene amplification and chromosomal rearrangements are frequent properties of cancer cells, provoking considerable interest in the mechanism of gene amplification and its consequences - particularly its relationship to chromosomal rearrangements. We recently studied the amplification of the gene for adenylate deaminase 2 (AMPD2) in Chinese hamster cells. Using fluorescent in situ hybridization (FISH), we found that early amplification of the AMPD2 gene is based on unequal gene segregation at mitosis, rather than local over-replication. We observed large inverted repeats of the amplified sequences, consistent with an amplification mechanism involving cycles of chromatid breakage, followed by fusion after replication and, in mitosis, the formation of bridges between the fused sister chromatids that leads to further breaks - a process we refer to as chromatid breakage-fusion-bridge (BFB) cycles. Our previous work left open the question of how this mechanism of gene amplification is related, if at all, to the chromosomal rearrangements that generate the dicentric, ring and double minute (DM) chromosomes observed in some AMPD2-amplified metaphase cells, which are not predicted intermediates of chromatid BFB cycles, although they could be generated by related chromosome BFB cycles. RESULTS: We have addressed this question using FISH with probes for the AMPD2 gene and other markers on the same chromosome. Our results are not consistent with the chromosome BFB cycle mechanism, in which two chromatids break simultaneously and fuse to generate, after replication, a dicentric chromosome. Rather, they suggest that dicentric chromosomes are generated by secondary events that occur during chromatid BFB cycles. Our results also suggest that DM chromosomes are generated by the 'looping-out' of a chromosomal region, generating a circular DNA molecule lacking a centromere; in this case, gene amplification would result from the unequal segregation of DM chromosomes at mitosis. CONCLUSION: We conclude that, at early stages of AMPD2 gene amplification, chromatid BFB cycles are a major source of both 'intrachromosomal' gene amplification and genomic rearrangement, which are first limited to a single chromosome but which can then potentially spread to any additional chromosome. It also seems that, occasionally, a DNA sequence including the AMPD2 gene can be excised, generating a DM chromosome and thus initiating an independent process of 'extrachromosomal' amplification. PMID- 15335746 TI - ATP induces large quaternary rearrangements in a cage-like chaperonin structure. AB - BACKGROUND: The chaperonins, a family of molecular chaperones, are large oligomeric proteins that bind nonnative intermediates of protein folding. They couple the release and correct folding of their ligands to the binding and hydrolysis of ATP. Chaperonin 60 (cpn60) is a decatetramer (14-mer) of 60 kD subunits. Folding of some ligands also requires the cooperation of cpn10, a heptamer of 10 kD subunits. RESULTS: We have determined the three-dimensional arrangements of subunits in Rhodobacter sphaeroides cpn60 in the nucleotide-free and ATP-bound forms. Negative stain electron microscopy and tilt reconstruction show the cylindrical structure of the decatetramer comprising two rings of seven subunits. The decatetramer consists of two cages joined base-to-base without a continuous central channel. These cages appear to contain bound polypeptide with an asymmetric distribution between the two rings. The two major domains of each subunit are connected on the exterior of the cylinder by a narrower bridge of density that could be a hinge region. Binding of ATP to cpn60 causes a major rearrangement of the protein density, which is reversed upon the hydrolysis of the ATP. Cpn10 binds to only one end of the cpn60 structure and is visible as an additional layer of density forming a cap on one end of the cpn60 cylinder. CONCLUSIONS: The observed rearrangement is consistent with an inward 5-10 degrees rotation of subunits, pivoting about the subunit contacts between the two heptamers, and thus bringing cpn60 domains towards the position occupied by the bound polypeptide. This change could explain the stimulation of ATPase activity by ligands, and the effects of ATP on lowering the affinity of cpn60 for ligands and on triggering the release of folding polypeptides. PMID- 15335747 TI - Genomic imprinting: imprints on islands. PMID- 15335748 TI - Virus-ligand structures: the viral canyon. PMID- 15335749 TI - Co-receptor selection: chance or necessity? PMID- 15335751 TI - Ecosystem dynamics: nature's short, sharp shocks. PMID- 15335750 TI - Neurodegeneration: great minds think alike. PMID- 15335752 TI - Cell-cycle control: turning on mitosis. PMID- 15335753 TI - Non-classical MHC molecules: the first line of defence? PMID- 15335754 TI - Nuclear pore structure: fingers in the pore. PMID- 15335755 TI - Cognition: dyslexic neural networks. PMID- 15335756 TI - Protein phosphorylation: phosphoprotein talk. PMID- 15335757 TI - Vertebrate development: factoring in the limb. PMID- 15335759 TI - X-linked immunodeficiency: B-cell diseases. PMID- 15335758 TI - Optical tweezers: Glasperlenspiel II. PMID- 15335760 TI - Receptor diversity: more serotonin receptors? PMID- 15335761 TI - Gene regulation: able assistants for homeodomain proteins. PMID- 15335762 TI - Cytoskeleton: dynamic actin. PMID- 15335763 TI - Flying kiwis and pattern information in biogeographic hypotheses. PMID- 15335765 TI - The connectional organization of neural systems in the cat cerebral cortex. AB - BACKGROUND: The mammalian brain consists of the cerebral cortical sheet, which is composed of many distinct areas, the cerebellar cortex, and many non-cortical nuclei. Powerful neuroanatomical techniques have revealed a large number of connections between these structures. The large number of brain structures and the very many connections between them form a strikingly complex network. The complexity of this network has made it difficult to understand how the central nervous system is organized. Recently, however, optimization analysis of an important subset of central nervous connections that occur between the different areas of the cerebral cortex has produced understandable and quantitative representations of the organization of cortical systems of the primate brain. RESULTS: Here we briefly report the extension of this approach to the cortical systems of the cat. There were four connectional clusters of cortical areas in the cat. These clusters of areas corresponded to the visual, auditory, and somato motor systems, and to the frontal and limbic areas, which we call the fronto limbic complex. All the major sensory systems were hierarchically organized, and their 'higher' stations were more closely associated with the fronto-limbic complex than were their 'lower' stations. CONCLUSIONS: Features of the organization of the cat brain, together with earlier primate results, suggest that there may be a common cortical plan in mammals. We suggest that this common plan may involve relatively discrete, hierarchically organized, cortical sensory systems and a topologically central fronto-limbic complex. Specific variations on this wiring plan may relate to evolutionary history and selection for particular ecological niches. PMID- 15335766 TI - Neural development: crest destiny. PMID- 15335767 TI - Enzyme catalysis: now you see it, now you don't. PMID- 15335769 TI - Opioid receptors: pipe dreams realized. PMID- 15335768 TI - Microtubule dynamics: severing microtubules in mitosis. PMID- 15335770 TI - Molecular evolution: slippery introns and globin gene evolution. PMID- 15335771 TI - Gene expression: chromatin and transcription factors: who's on first? PMID- 15335772 TI - Disease resistance: good news from Gambia. PMID- 15335773 TI - Colour vision: human V4? PMID- 15335774 TI - Vesicle traffic: G whizz. PMID- 15335775 TI - Protein folding: junior chaperones. PMID- 15335776 TI - Vision: insights into blindsight? PMID- 15335777 TI - Acetylcholine receptors: peering down a pore. PMID- 15335778 TI - X-inactivation: controlling the X chromosome. PMID- 15335779 TI - Myosin superfamily: a multitude of myosins. PMID- 15335780 TI - Recurring structural motifs in proteins with different functions. AB - BACKGROUND: Structures that have diverged from a common ancestor often retain functional and sequence similarity, although the latter may be very reduced. Even so, the overall fold of the structure is generally highly conserved. Now however, several have been identified of proteins that have been identified that have different functions but which have converged to a similar fold. These proteins will also have low sequence identities. RESULTS: By comparing the complete structure databank against itself, using sequence and structure alignment techniques, we have been able to identify six new examples of structurally related folds that have no apparent sequence or functional similarity. These related proteins include a family of crambin-like folds and a family of ferredoxin II folds. We found that all the similarities between structures are present in small proteins and occur as motifs within the core of a larger protein. CONCLUSION: The low sequence similarity and the lack of any obvious functional relationship between proteins with similar structures suggest that the proteins have diverged from independent ancestors. The similarities may therefore be of interest for understanding the various stereochemical and physical criteria that operate to generate a favourable fold. PMID- 15335782 TI - Viewpoint: microsatellites and the new genetic maps. PMID- 15335781 TI - Structural similarity of DNA-binding domains of bacteriophage repressors and the globin core. AB - BACKGROUND: In recent years, the determination of large numbers of protein structures has created a need for automatic and objective methods for the comparison of structures or conformations. Many protein structures show similarities of conformation that are undetectable by comparing their sequences. Comparison of structures can reveal similarities between proteins thought to be unrelated, providing new insight into the interrelationships of sequence, structure and function. RESULTS: Using a new tool that we have developed to perform rapid structural alignment, we present the highlights of an exhaustive comparison of all pairs of protein structures in the Brookhaven protein database. Notably, we find that the DNA-binding domain of the bacteriophage repressor family is almost completely embedded in the larger eight-helix fold of the globin family of proteins. The significant match of specific residues is correlated with functional, structural and evolutionary information. CONCLUSION: Our method can help to identify structurally similar folds rapidly and with high-sensitivity, providing a powerful tool for analyzing the ever-increasing number of protein structures being elucidated. PMID- 15335783 TI - Cell polarity: barriers to diffusion. PMID- 15335784 TI - Mating systems: save the male. PMID- 15335785 TI - Protein modules: structural convergence and horizontal transfer? PMID- 15335786 TI - Vision: new perspectives on binocular stereopsis? PMID- 15335787 TI - Cell cycle: bring on the phosphatases. PMID- 15335788 TI - Cytotoxic mechanisms: cytolytic T cells keep their secrets. PMID- 15335789 TI - Gonad development: inhibiting inhibin. PMID- 15335790 TI - Iron-sulphur proteins: crosslinked by a cluster. PMID- 15335791 TI - Protein translocation: through the mitochondrial inner membrane. PMID- 15335792 TI - Evolutionary genetics: signs of selection in silent substitutions. PMID- 15335793 TI - Cortical circuitry: filling in the gaps. PMID- 15335794 TI - Protein-DNA interactions: how GCN4 binds DNA. PMID- 15335795 TI - Differentiation therapy: retinoic acid-induced remission. PMID- 15335796 TI - Extracellular matrix: the thrombospondin family. PMID- 15335797 TI - HOM/Hox genes of Artemia: implications for the origin of insect and crustacean body plans. AB - BACKGROUND: Insects and crustaceans are generally assumed to derive from a segmented common ancestor that had a distinct head but uniform, undifferentiated trunk segments. The subdivision of the body into functionally distinct regions (e.g. thorax and abdomen) is thought to have evolved independently in these two lineages. In insects, the differences between segments in the trunk are controlled by the Antennapedia-like genes of the homeotic gene clusters. Study of these genes in crustaceans should provide a basis for comparing body plans and assessing their evolutionary origin. RESULTS: Using a polymerase chain reaction (PCR) / inverse PCR strategy, we have isolated six genes of the HOM/Hox family from the crustacean Artemia franciscana. Five of these are clearly identifiable as specific homologues of the insect homeotic genes Dfd, Scr, Antp, Ubx and abdA. The sixth appears to have no close counterpart in insects. CONCLUSION: All the homeotic genes that specify middle body regions in insects originated before the divergence of the insect and crustacean lineages, probably not later than the Cambrian (about 500 million years ago). A commonly derived groundplan may underlie segment diversity in these two groups. PMID- 15335798 TI - Why physicists like models and why biologists should. PMID- 15335799 TI - Colour vision: mixing genes and mixing colours. PMID- 15335800 TI - Prokaryotic secretion: a signal recognition particle in Escherichia coli? PMID- 15335801 TI - Flower development: manipulating floral organ identity. PMID- 15335802 TI - DNA topology: untangling the steps in chromosome segregation. PMID- 15335804 TI - Human brain: visualizing the motion area. PMID- 15335803 TI - T-cell activation: ZAPping the T-cell receptor. PMID- 15335805 TI - Evolutionary ecology: the ecology of evolutionary succession. PMID- 15335806 TI - Muscular dystrophy: dystrophin and partners at the cell surface. PMID- 15335807 TI - DNA-binding proteins: saddling up the TATA box. PMID- 15335808 TI - Helper T cells: polarized patterns of cytokine secretion. PMID- 15335809 TI - Cystic fibrosis: ion channels and transmembrane transporters. PMID- 15335811 TI - Cardiovascular disease: shuffling the cards. PMID- 15335810 TI - Evolution: a lac of interest in evolution. PMID- 15335812 TI - Protein turnover: proteasome location. PMID- 15335813 TI - Association of pRas and pRaf-1 in a complex correlates with activation of a signal transduction pathway. AB - BACKGROUND: A key pathway for transduction of proliferative, developmental and oncogenic stimuli from receptors at the cell surface to transcription factors located in the nucleus involves the activation of pRas and pRaf-1. Recent publications have described a physical interaction between pRas and pRaf-1, either as ectopic proteins in yeast or as recombinant proteins added to cellular extracts. Until now, however, physical complexes that include pRas and pRaf-1 have not been identified as native structures in mammalian cells. RESULTS: We have directly identified a pRas-pRaf-1 complex in extracts of mammalian cells. Formation of the complex is augmented in neoplastically transformed cells expressing constitutively activated pRas. Moreover, the complexes form in concert with the activation of pRas during intracellular signalling through the T-cell receptor in T-leukemia cells. CONCLUSIONS: We propose that, pRas signals to pRaf 1 in vivo by means of a direct physical interaction that results in activation of the pRaf-1 protein kinase. PMID- 15335814 TI - Characterization of PKC2, a gene encoding a second protein kinase C isotype of Saccharomyces cerevisiae. AB - BACKGROUND: Protein kinase C (PKC) has attracted considerable attention over the past decade, primarily because of its presumed role in cellular growth control and tumourigenesis. Mammalian cells express at least 10 different isozymes of PKC; it is this complexity that has made elucidating the precise functions of PKC: so difficult. The identification of PKC homologues in organisms such as Drosophila, Xenopus, Dictyostelium, Aplysia and Caenorhabditis indicates that the enzyme is evolutionarily conserved, and this has stimulated our search for counterparts in the yeast Saccharomyces cerevisiae, in which powerful genetic analyses can be used. To date, only one PKC homologue, PKC1, has been identified in yeast and no biochemical activity has been definitively ascribed to the encoded protein. This, and the inability to identify other PKC homologues in yeast by DNA hybridization, has led to doubts about the existence of PKC isozymes in yeast. We have taken the approach of screening yeast expression libraries with anti-PKC antibodies in an attempt to identify further homologues. RESULTS: We have identified a novel PKC isozyme, Pkc2p, encoded by the gene PKC2. We report here the sequence of PKC2 and a comparison showing its similarity to other PKCs. Phylogenetic analysis suggests that all known PKC genes, including PKC2, originated from a common ancestor. Disruption of the PKC2 protein-coding region, deleting the entire catalytic domain of the encoded enzyme, is not lethal to yeast growing on rich media. However, the pkc2 mutant, unlike wild-type strains, fails to grow on minimal media containing limited concentrations of amino acids. This implicates Pkc2p in the response of yeast cells to amino-acid starvation. CONCLUSION: We have shown that yeast cells do express more than one PKC isozyme, by identifying and characterizing a novel PKC gene PKC2, the product of which may be involved in the cellular response to amino-acid starvation. PMID- 15335815 TI - Transcriptional interactions of transforming growth-factor-beta with pro inflammatory cytokines. AB - BACKGROUND: Inflammation and tissue injury are characterized by a massive infiltration of mononuclear cells. These pro-inflammatory cells, which are the precursors of an inflammatory response by the immune system, secrete a variety of cytokines and growth factors that alter the biosynthetic repertoire of the resident connective tissue cells. Specifically, expression of connective tissue matrix metalloproteinases, such as stromelysin and interstitial collagenase, is enhanced, together with the expression of chemoattractants for leukocytes, such as interleukin-8 (IL-8). These events lead to increased connective tissue degradation. We have examined the growth factor regulation of expression in cultured fibroblasts of the prototypic pro-inflammatory factors interstitial collagenase and IL-8. RESULTS: We demonstrate that transforming growth factor beta (TGF-beta) does not interfere with cytokine-induced IL-8 gene expression, nor does it affect the activity of NF-kappaB-driven promoters. In contrast, TGF beta down-regulates collagenase gene expression through the induction of the jun B proto-oncogene. Jun-B is a negative regulator of c-jun, which mediates cytokine activation of collagenase gene expression through its action as a component of the AP-1 transcription factor. CONCLUSION: Our data suggest that TGF-beta may attenuate the deleterious events that occur in inflammation by preventing cytokine-induced extracellular matrix degradation, although it does not affect cytokine-induced recruitment of pro-inflammatory cells. Furthermore, our data suggest a potential therapeutic use for jun-B, which may be a candidate for gene therapy in disease states that are characterized by excessive connective tissue degradation. PMID- 15335816 TI - A common rule governs the synaptic locus of both short-term and long-term potentiation. AB - BACKGROUND: At synapses between neurons in the brain, transmitter molecules are released from presynaptic terminals in multi-molecular packets called quanta. Excitatory synapses in the CA1 region of the hippocampus show a long-lasting increase in strength known as long-term potentiation (LTP), which may be important for some kinds of learning and memory. LTP can involve an increase in the number of quanta released, or in the size of the response each quantum produces in the postsynaptic cell, or both, depending on the initial condition of the synapse. These synapses also show two forms of brief potentiation: post tetanic potentiation (PTP), which lasts for a minute or less and involves only modifications at the presynaptic terminal, and short-term potentiation (STP), which lasts rather longer. The significance of STP, the mechanisms whereby it is produced and its relationship to other forms of potentiation are poorly understood. We have studied STP electrophysiologically using slices of the rat hippocampus maintained in vitro. RESULTS: We found that STP, like LTP, can involve increases in either the number of quanta released, or their postsynaptic effect, or both. The rule governing the relative contribution from these two mechanisms appears to be the same as operates during LTP. Both the presynaptic and postsynaptic changes can develop equally rapidly and so must involve fast acting messenger systems. CONCLUSIONS: STP seems to be a separate phenomenon from PTP, but appears closely related to LTP. The rapidity of its onset may require a reappraisal of current understanding of the messenger systems involved in bringing about changes in synaptic strength. PMID- 15335817 TI - A new method for manipulating transgenes: engineering heat tolerance in a complex, multicellular organism. AB - BACKGROUND: Heat-shock proteins (hsps) are thought to protect cells against stresses, especially due to elevated temperatures. But while genetic manipulation of hsp gene expression can protect microorganisms and cultured metazoan cells against lethal stress, this has so far not been demonstrated in multicellular organisms. Testing whether expression of an hsp transgene contributes to increased stress tolerance is complicated by a general problem of transgene analysis: if the transgene cannot be targeted to a precise site in the genome, newly observed phenotypes may be due to either the action of the transgene or mutations caused by the transgene insertion. RESULTS: To study the relationship between heat tolerance and hsp expression in Drosophila melanogaster, we have developed a novel method for transgene analysis, based upon the site-specific FLP recombinase. The method employs site-specific sister chromatid exchange to create an allelic series of transgene insertions that share the same integration site, but differ in transgene copy number. Phenotypic differences between members of this series can be confidently attributed to the transgenes. Using such an allelic series and a novel thermotolerance assay for Drosophila embryos, we investigated the role of the 70 kD heat-shock protein, Hsp 70, in thermotolerance. At early embryonic stages, Hsp70 accumulation was rate-limiting for thermotolerance, and elevated Hsp70 expression increased survival at extreme temperatures. CONCLUSION: Our results provide an improved method for analyzing transgenes and demonstrate that, in Drosophila, Hsp70 is a critical thermotolerance factor. They show, moreover, that manipulating the expression of a single hsp can be sufficient to improve the stress tolerance of a complex multicellular organism. PMID- 15335818 TI - Peptide selection by class I molecules of the major histocompatibility complex. AB - Class I molecules of the major histocompatibility complex (MHC) bind peptides derived from cytoplasmic proteins. Comparison of over 100 such peptides reveals the importance of the carboxy-terminal residue in selective binding. Recent evidence implicates the proteases and transporters of the processing pathway in providing peptides with the correct residues at the carboxyl terminus. PMID- 15335819 TI - Patterning the Arabidopsis root. PMID- 15335820 TI - Insights from inducible membranes. PMID- 15335821 TI - From EF-Tu to p21ras and back again. PMID- 15335822 TI - A boom time for necrobiology. PMID- 15335823 TI - Promoting motor neuron survival. PMID- 15335824 TI - Hanging nests on a phylogenetic tree. PMID- 15335825 TI - A chaperone with a sweet tooth. PMID- 15335827 TI - Going down the way you came up. PMID- 15335826 TI - The secrets of severing? PMID- 15335828 TI - Conducting the G-quartet. PMID- 15335829 TI - Jamming for sex. PMID- 15335830 TI - Spurring on transcription? PMID- 15335831 TI - How does the immune system remember? PMID- 15335832 TI - Sleeping neuroblasts. PMID- 15335833 TI - Two steps easier than one? PMID- 15335834 TI - Keeping the stores full. PMID- 15335835 TI - Riding the evanescent wave. PMID- 15335836 TI - Monocyte haptotaxis induced by the RANTES chemokine. AB - BACKGROUND: Soluble mediators and inducible cell-surface molecules coordinate the ordered cascade of events giving rise to inflammation. The specific mechanisms underlying the attraction of antigen-specific cells into a site of inflammation remain sketchy, however. In particular, it is unclear how chemoattractants cause rapidly moving immune cells to adhere to the blood vessel wall and to enter inflamed tissues. RESULTS: Here we show that RANTES, a potent chemo-attractant for monocytes and T lymphocytes, is inducibly expressed within an inflamed organ, binds to endothelial cells, and promotes haptotaxis, the migration of cells induced by surface-bound gradients. CONCLUSION: These findings lead us to propose a model for the role of RANTES in the migration of antigen-specific immune cells into an inflammatory site. PMID- 15335837 TI - Automatic and accurate method for analysis of proteins that undergo hinge mediated domain and loop movements. AB - BACKGROUND: The structures of proteins that undergo significant main-chain conformational change are reported with increasing frequency. Three-dimensional atomic models are often available for two alternative conformational states of the same molecule. Inspection has shown these states to be varied in nature, arising by mechanisms that include hinge-facilitated closure between domains and smaller-scale loop motions within domains; these movements are often induced by metal ion binding or ligand binding. Polypeptides that display flexible segments are also observed in different crystal conformations or as alternatively packed subunits. Although subjective visual inspection has been previously used to compare structures and analyze conformational changes, there is a need for an objective method. RESULTS: We have developed a straightforward, robust, and objective algorithm that can locate the residues that mediate and participate in the changes between the two conformational states. Our method does not require initial superpositioning. We illustrate the method by considering several test cases. The first example is maltose binding protein, a polypeptide that exhibits rigid-body domain closure involving multiple hinges. The second is lactate dehydrogenase, which undergoes both loop and subdomain movement; we accurately describe the location and relative magnitude of these deformations. Finally, in the example of aspartate transcarbamoylase, both hinge-mediated domain movement and functionally relevant loop rearrangements are described. In the instances in which domain closure occurs, the residues that serve as hinges between the domains involved are accurately predicted. In addition, our technique successfully identifies the exact residues that undergo intra-domain loop movements, even for movements that are accompanied by larger scale inter-domain rearrangements. CONCLUSIONS: Our algorithm is successful in its comprehensive analysis and description of complex hinge-mediated domain motion for all structures displaying rigid-body movement and is accurate in identifying the location of any independent intra-domain rearrangements. PMID- 15335838 TI - Neurotransmitter release regulated by nitric oxide in PC-12 cells and brain synaptosomes. AB - BACKGROUND: Nitric oxide is a messenger molecule of the nervous system, which is produced by the enzyme nitric oxide synthase, which may regulate cyclic guanosine monophosphate levels and which has been implicated in the control of neurotransmitter release. PC-12 pheochromocytoma cells differentiate to form neuronal cells in culture when they are exposed to nerve growth factor. The levels of cyclic guanosine monophosphate in the cells and their ability to release acetylcholine in response to K(+)-depolarization are both maximal after eight days of treatment with nerve growth factor. We set out to assess a possible role for nitric oxide in the processes that occur in differentiating PC-12 cells. RESULTS: Nitric oxide synthase is first evident in differentiating PC-12 cells eight days after beginning treatment with nerve growth factor, coinciding with the marked increase in K(+)-depolarization-induced release of acetylcholine. The release of both acetylcholine and dopamine in response to K(+)-depolarization is blocked by inhibitors of nitric oxide synthase and by hemoglobin, which binds nitric oxide. Providing l-arginine, a precursor required for nitric oxide synthesis, reverses the effects of the inhibitors. In synaptosomal preparations from the corpus striatum, inhibitors of nitric oxide synthase prevent the release of glutamate in response to the glutamate derivative N-methyl-d-aspartate but not in response to K(+)-depolarization. CONCLUSION: Nitric oxide may mediate the release of acetylcholine and dopamine in response to K(+)-depolarization in PC-12 cells and the release of glutamate in response to N-methyl-d-aspartate in striatal synaptosomes. Nitric oxide synthase expression is induced after eight days of treating PC-12 cells with nerve growth factor, coinciding with a marked enhancement of the release of neurotransmitters in response to K(+) depolarization. PMID- 15335839 TI - Distortion in those good vibrations. PMID- 15335840 TI - Keep the noise down. PMID- 15335841 TI - Falling off the knife edge. PMID- 15335842 TI - The whole body view. PMID- 15335843 TI - Anti-Ras drugs come of age. PMID- 15335844 TI - Dynamin, SH3 domains and endocytosis. PMID- 15335845 TI - Fitting the bill. PMID- 15335846 TI - Seeing the light through the blobs. PMID- 15335847 TI - T-cell differentiation under control. PMID- 15335848 TI - An expanding story. PMID- 15335849 TI - Segmentation moves to the fore. PMID- 15335850 TI - A model catalyst of protein disulphide bond formation. PMID- 15335852 TI - Why species and subspecies? PMID- 15335851 TI - Inositol lipid signalling exacts a transfer fee. PMID- 15335853 TI - Linear and non-linear mechanisms in pattern vision. PMID- 15335854 TI - Transcytosis of the polymeric immunoglobulin receptor in cultured hippocampal neurons. AB - BACKGROUND: A wide variety of proteins are transported across epithelial cells by vesicular carriers. This process, transcytosis, is used to generate cell surface polarity and to transport macromolecules between the luminal and serosal sides of the epithelial layer. The polymeric immunoglobulin receptor is a well characterized transcytotic molecule in epithelia. It binds to its ligand, polymeric immunoglobulin, at the basolateral surface, and the receptor-ligand complex is transcytosed to the apical surface, where the ligand is released. Our previous studies have shown that hippocampal neurons may employ mechanisms similar to those of epithelial cells to sort proteins to two plasma membrane domains. The machinery used for axonal delivery recognizes proteins that are targeted apically in epithelia, whereas basolaterally destined proteins are delivered to the dendrites. It has not been clear, however, whether transcytosis occurs in neurons. RESULTS: We report expression of the polymeric immunoglobulin receptor in cultured hippocampal neurons, using a Semliki Forest Virus expression system, and show by immunofluorescence microscopy that the newly synthesized receptor is targeted from the Golgi complex predominantly to the dendrites - only about 20% of the infected neurons display axonal immunofluorescence. Addition of ligand leads to significant redistribution of the receptor to the axons, shown by an approximately three-fold increase in axonal immunoreactivity with the anti receptor antibodies. CONCLUSIONS: Our results suggest that a transcytotic route, analogous to that in epithelia, exists in neurons, where it transports proteins from the somatodendritic to the axonal domain. Cultured neurons expressing the polymeric immunoglobulin receptor offer an experimental system that should be useful for further characterization of this novel neuronal pathway at the molecular and functional level. PMID- 15335855 TI - B-cell antigen receptor motifs have redundant signalling capabilities and bind the tyrosine kinases PTK72, Lyn and Fyn. AB - BACKGROUND: The 13 cell antigen receptor (BCR) is a multimeric protein complex consisting of an antigen recognition structure (membrane immunoglobulin) and two associated proteins, lg-alpha and Ig-beta It has been proposed that signalling through the BCR involves Ig-alpha and Ig-beta. Both of these proteins contain within their cytoplasmic domains an amino-acid motif that is present in a number of immune recognition receptors, including the BCR, T-cell antigen receptor and Fc receptor complexes. This motif, termed the antigen-receptor homology motif (ARH1), appears to have signal transduction ability. RESULTS: We now show that the presence of cytoplasmic regions containing the ARM motif from either Ig-alpha or Ig-beta is sufficient to confer signalling capability on an otherwise non functional fusion protein. Both Ig-alpha- and Ig-beta-containing chimeras induced, in an apparently redundant fashion, signalling events seen upon membrane immunoglobulin crosslinking, including tyrosine phosphorylation of particular proteins, phosphoinositicle breakdown and calcium mobilization. Furthermore, crosslinking of the chimeras resulted in tyrosine phosphorylation of the Ig-alpha and Tg-beta tails and their association with the tyrosine kinases PTK72, p53/56(lyn) and p59(fyn). CONCLUSIONS: These observations indicate that Ig-alpha and Ig-beta are responsible for coupling membrane immunoglobulin to intracellular signalling components. Moreover, they demonstrate that a number of tyrosine kinases associate directly with the cytoplasmic domains of both Ig-alpha and Ig beta. Stimulation of the chimeras, which results in tyrosine phosphorylation of the ig-alpha and Ig-beta tails, is a prerequisite for some of these associations. The implications of these findings for the mechanism by which the BCR initiates the signalling reactions are discussed. PMID- 15335856 TI - Peptide 'Velcro': design of a heterodimeric coiled coil. AB - BACKGROUND: The leucine zipper is a protein structural motif involved in the dimerization of a number of transcription factors. We have previously shown that peptides corresponding to the leucine-zipper region of the Fos and Jun oncoproteins preferentially form heterodimeric coiled coils, and that simple principles involving electrostatic interactions are likely to determine the pairing specificity of coiled coils. A critical test of these principles is to use them as guidelines to design peptides with desired properties. RESULTS: Based on studies of the Fos, Jun and GCN4 leucine zippers, we have designed two peptides that are predominantly unfolded in isolation but which, when mixed, associate preferentially to form a stable, parallel, coiled-coil heterodimer. To favor heterodimer formation, we chose peptide sequences that would be predicted to give destabilizing electrostatic interactions in the homodimers that would be relieved in the heterodimer. The peptides have at least a 10(5)-fold preference for heterodimer formation, and the dissociation constant of the heterodimer in phosphate-buffered saline is approximately 30 nM at pH 7 and 20 degrees C. Studies of the pH and ionic strength dependence of stability confirm that heterodimer formation is favored largely as a result of electrostatic destabilization of the homodimers. CONCLUSIONS: Our successful design strategy supports previous conclusions about the mechanism of interaction between the Fos and Jun oncoproteins. These results have implications for protein design, as they show that it is possible to design peptides with simple sequences that have a very high preference to pair with one another. Finally, these sequences with 'Velcro'-like properties may have practical applications, including use as an affinity reagent, in lieu of an epitope tag, or as a way of bringing together two molecules in a cell. PMID- 15335857 TI - Olfactory receptors. PMID- 15335858 TI - Architectural elements in nucleoprotein complexes. PMID- 15335859 TI - Shedding light on the translocation pore. PMID- 15335860 TI - A 'roll' in acute inflammation. PMID- 15335861 TI - Arresting G-protein coupled receptor activity. PMID- 15335862 TI - Here today, bone tomorrow. PMID- 15335863 TI - Turn-of-the-century electron microscopy. PMID- 15335864 TI - Stop that cell cycle. PMID- 15335865 TI - Scrapie agent reflication without the prion protein? PMID- 15335866 TI - VEGF/VPF: the angiogenesis factor found? PMID- 15335867 TI - The problems of building a head. PMID- 15335868 TI - Trans-plication factors? PMID- 15335869 TI - Signaling the insulin receptor way. PMID- 15335870 TI - Does degradation lead to segregation? PMID- 15335871 TI - RNA zipcodes for cytoplasmic addresses. PMID- 15335872 TI - A community effect in muscle development. AB - BACKGROUND: Most vertebrate tissues arise by embryonic induction, as a result of which new cell layers are formed. These are subsequently subdivided into discrete groups of homogeneous cell populations, each containing different cell-types with specific gene expression. There is preliminary evidence from previous work that the mesoderm-forming induction in amphibian development may be followed by a further interaction among some of the induced mesoderm cells, and that this could be required for muscle gene activation in uniform cell populations. RESULTS: We have established the existence, time and place of this further cell interaction by transplanting muscle progenitor cells from Xenopus mid-gastrulae into ectoderm sandwiches, and then culturing these constructs until the time of muscle gene activation. We find that cells implanted as reaggregates, but not those implanted as single cells, activate early myogenic genes and later muscle-specific genes. More than 100 cells must be near each other for muscle gene activation. These cells can induce non-muscle mesoderm cells to express muscle genes by emitting a signal that differs from the preceding mesoderm induction signal. Muscle gene activation under these conditions does not require gap junction communication. CONCLUSION: Cells within the muscle progenitor region of a Xenopus embryo need to interact with each other in order to activate muscle genes in homogeneous cell groups. This exemplifies the 'community effect', which may be a widespread developmental mechanism used to increase the homogeneity within, and demarkation between, embryonic tissues. PMID- 15335874 TI - Identifying the three signals. PMID- 15335873 TI - Mitotic regulation of protein phosphatases by the fission yeast sds22 protein. AB - BACKGROUND: Cell cycle progression requires the activity of protein kinases and phosphatases at critical points in the cell cycle in all eukaryotes. We have previously reported that the dis2(+) and sds2(+) genes of fission yeast encode redundant catalytic subunits of a type 1-like protein phosphatase. The sds22(+) gene was shown to be essential for cell viability and to interact genetically with dis2(+) and sds21(+). RESULTS: Here we show by immunoprecipitation that the sds22 protein physically interacts with the dis2 and sds21 proteins, and that sds22-associated phosphatase activity has altered substrate specificity, The loss of sds22 function by a temperature sensitive mutation leads to cell cycle arrest at mid-mitosis, at which point cdc2-dependent histone Hl kinase activity is high while sds22-dependent H1 phosphatase activity is low. To examine the unusual properties of sds22 protein structure, we analyzed a collection of sds22 deletion and point mutants by a variety of functional criteria. CONCLUSION: We propose that sds22 is a regulatory subunit of the dis2/sds21 phosphatase catalytic subunits and that sds22-bound phosphatase carries a key phosphatase activity essential for the progression from metaphase to anaphase. Mutational analysis indicates that dis2/sds21 interacts with the central repetitive domain of sds22, while the C-terminal and central regions of sds22 may be involved in subcellular targeting and the N-terminus is important for stability. PMID- 15335875 TI - Taking the Rabs off endocytosis. PMID- 15335876 TI - Two fingers for membrane traffic. PMID- 15335877 TI - Which were the real New Zealanders? PMID- 15335878 TI - The ear's fast cellular motor. PMID- 15335879 TI - A new use for an old enzyme. PMID- 15335880 TI - Modules for pattern recognition. PMID- 15335882 TI - Ants with a taste for millipedes. PMID- 15335881 TI - Immortal germ cells? PMID- 15335883 TI - A new break in V(D)J recombination. PMID- 15335884 TI - Long lost cousins of actin. PMID- 15335886 TI - Loading synaptic vesicles with neurotransmitter. PMID- 15335885 TI - Haldane's rule and the hazards of heterogamety. PMID- 15335887 TI - A glimpse into the spliceosome. PMID- 15335888 TI - GTPase enters the ring. PMID- 15335890 TI - Cortical connections in cocultures. PMID- 15335889 TI - Stranded in an active gene. PMID- 15335891 TI - Unfaithful and asymmetric sex. PMID- 15335892 TI - Marked by a mucin. PMID- 15335893 TI - The highs and lows of an NGF receptor. PMID- 15335894 TI - Crossing the threshold. PMID- 15335895 TI - Spindle positioning and cell polarity. PMID- 15335896 TI - Hyperacuity of those in the know. PMID- 15335897 TI - Homologue sweet homologue. PMID- 15335899 TI - It's deja vu all over again. PMID- 15335898 TI - TCP1 - molecular chaperonin of the cytoplasm? PMID- 15335900 TI - Essential tools for viral transmission? PMID- 15335901 TI - No pain, no gain. PMID- 15335902 TI - Wither the beauty's transient flower. PMID- 15335903 TI - Soluble factors and solid phases. PMID- 15335904 TI - Fabulous Fabs from phage. PMID- 15335905 TI - More than one way to slice an egg. PMID- 15335906 TI - Potentially right on both sides. PMID- 15335907 TI - Fishing for partners. PMID- 15335908 TI - When is a species not a species? PMID- 15335909 TI - Frozen stiff. PMID- 15335911 TI - Safe havens for self-reactive cells. PMID- 15335910 TI - Transporters of delight. PMID- 15335912 TI - A switch-hitter at the Start of the cell cycle. PMID- 15335913 TI - Guide RNA tails of the unexpected. PMID- 15335915 TI - Not so groovy after all? PMID- 15335914 TI - Elastic nets, travelling salesmen and cortical maps. PMID- 15335916 TI - New ribozymes for old reactions. PMID- 15335917 TI - A new signalling role for salicylic acid. PMID- 15335918 TI - A function for ribophorins. PMID- 15335919 TI - A gene for language? PMID- 15335920 TI - Homing in on an endosymbiotic endonuclease. PMID- 15335921 TI - Resting places on folding pathways. PMID- 15335922 TI - Protein folding by stages. PMID- 15335923 TI - Quest's end and questions' beginning. PMID- 15335924 TI - Down the slippery slope. PMID- 15335925 TI - Finding the stepping stones downstream of Ras. PMID- 15335926 TI - The case of the missing CD4s. PMID- 15335927 TI - Picking the needle from the haystack. PMID- 15335928 TI - Quantized secretion of hormones. PMID- 15335929 TI - On course for collision? PMID- 15335931 TI - Distinguishing self from non-self. PMID- 15335930 TI - Testing time for muscle. PMID- 15335932 TI - Keeping the viral coat on. PMID- 15335933 TI - When apoptosis fails. PMID- 15335934 TI - More than one way to bug a plant. PMID- 15335935 TI - Multifarious IP3 receptors. PMID- 15335936 TI - Altogether now - sequencing the yeast genome. PMID- 15335937 TI - Classical sex education. PMID- 15335938 TI - No common motif. PMID- 15335940 TI - Holding on and letting go. PMID- 15335939 TI - Turning vision into action. PMID- 15335941 TI - A worm's eye view of cell motility. PMID- 15335942 TI - How mycoplasma ring the changes. PMID- 15335944 TI - Three routes for receptor-mediated Ca2+ entry. PMID- 15335943 TI - Stutter, stumble, or fall of a kinase? PMID- 15335945 TI - A terminal case of modulation. PMID- 15335946 TI - Green ancestry of malarial parasites? PMID- 15335947 TI - A switch in time. PMID- 15335948 TI - Moving off the beaten track. PMID- 15335949 TI - Ras regulation: putting back the GTP. PMID- 15335950 TI - Why does the cortex oscillate? PMID- 15335952 TI - Bridging the gaps. PMID- 15335951 TI - Latest episodes in the Golgi serial. PMID- 15335953 TI - The lessons from the nematode. PMID- 15335954 TI - Down the tube. PMID- 15335955 TI - Coming in from the twilight zone. PMID- 15335956 TI - Repopulating the mortal brain with immortal cells. PMID- 15335957 TI - Distinctly different...or really much the same? PMID- 15335958 TI - Puzzles of chloroplast ancestry. PMID- 15335959 TI - Double or nothing. PMID- 15335960 TI - Fruitful outcomes of intracellular cross-talk. PMID- 15335961 TI - Phage power. PMID- 15335962 TI - Twisting into shape. PMID- 15335963 TI - Removing constraints on neural sprouting. PMID- 15335965 TI - Mouse map fills the gap. PMID- 15335964 TI - What does the locus control region control? PMID- 15335966 TI - A plant oncogene uncovered. PMID- 15335967 TI - Vagaries of vav. PMID- 15335969 TI - Virus structure puzzle solved. PMID- 15335968 TI - Dicing with darwin. PMID- 15335970 TI - Excitatory amino acids provide the stimulus. PMID- 15335971 TI - RAG knockouts deliver a one/two punch. PMID- 15335973 TI - Will the real splicing sites please light up? PMID- 15335972 TI - Angling for recognition. PMID- 15335974 TI - Microtubules on the move? PMID- 15335975 TI - Visual deficits in dyslexia? PMID- 15335977 TI - Adaptive chaos and AIDS. PMID- 15335976 TI - Nuclear PIPs. PMID- 15335978 TI - HMGs everywhere. PMID- 15335979 TI - Peptides tailored to perfection? PMID- 15335980 TI - Down, but not out for the count. PMID- 15335981 TI - Autoregulation through translation. PMID- 15335982 TI - Variations in retinal degenerations. PMID- 15335983 TI - A sense of smell. PMID- 15335984 TI - Won't you change partners and dance? PMID- 15335985 TI - Unzipping the secrets of coiled-coils. PMID- 15335986 TI - Telomerase and chromosome healing. PMID- 15335987 TI - nef-naf nexus? PMID- 15335988 TI - Flight of fancy or a major new school? PMID- 15335989 TI - Bones need SOFAs. PMID- 15335990 TI - Patterning the developing cerebral cortex. PMID- 15335991 TI - Antigens coming to a sticky end. PMID- 15335992 TI - Phage introns on the hop: an alternative view. PMID- 15335993 TI - The Yin and the Yang of mammalian transcription. PMID- 15335994 TI - From G minor to G major. PMID- 15335995 TI - Inching towards a vaccine for AIDS. PMID- 15335996 TI - Modes of transduction. PMID- 15335997 TI - Actin filaments on the move. PMID- 15335998 TI - Telomere chromatin and gene expression. PMID- 15335999 TI - NGF takes shape. PMID- 15336000 TI - Modelling chronic myelogenous leukemia. PMID- 15336001 TI - Molecular rulers in muscle? PMID- 15336003 TI - Body building exercises. PMID- 15336002 TI - Defining a new GAP family. PMID- 15336004 TI - Eyes do more than see. PMID- 15336005 TI - Sticking together for a difficult passage. PMID- 15336006 TI - bcl-2 to the rescue. PMID- 15336007 TI - Janus-faced proteins. PMID- 15336008 TI - Sex, nose and genotype. PMID- 15336009 TI - Kalinin, epiligrin and GB3 antigen: kalinepiligrinin-3? PMID- 15336011 TI - The platonic neuron gets the hots. PMID- 15336010 TI - Just say NO. PMID- 15336012 TI - Common denominators for sex. PMID- 15336013 TI - Signal transduction meets the secretory pathway. PMID- 15336014 TI - Crystallography fixes it for nitrogenase. PMID- 15336016 TI - A role for glia in LHRH release. PMID- 15336015 TI - Transforming the Hox code. PMID- 15336017 TI - Bacterial pathogens caught in the actin. PMID- 15336018 TI - Enigma variations. PMID- 15336019 TI - Switching partners? PMID- 15336020 TI - DNA between the sheets. PMID- 15336021 TI - Sight unseen? PMID- 15336022 TI - Rac and Rho in control. PMID- 15336023 TI - Activators, deactivators and deactivated activators. PMID- 15336024 TI - Sex in the soil. PMID- 15336025 TI - Anchored fore and aft. PMID- 15336026 TI - Unifying forces for chromosomes in mitosis. PMID- 15336027 TI - Hunting for the pore of voltage-gated channels. PMID- 15336028 TI - A division too far? PMID- 15336029 TI - Worrying about p53. PMID- 15336030 TI - Superperfect enzymes. PMID- 15336031 TI - Kinase cascade conserved. PMID- 15336032 TI - High fives or hand clasps? PMID- 15336033 TI - Life without water. PMID- 15336034 TI - Motion may be seen but not used. PMID- 15336035 TI - Missing links between receptors and Ras. PMID- 15336036 TI - The high genomic mutation rate. PMID- 15336037 TI - A better grip on calmodulin. PMID- 15336038 TI - An unexpected petition for pardon. PMID- 15336039 TI - More than just a membrane anchor. PMID- 15336040 TI - Two routes to neurotoxicity? PMID- 15336041 TI - The Pierian Spring. PMID- 15336042 TI - Why have Y chromosomes? PMID- 15336044 TI - A handy enzyme for transcription. PMID- 15336043 TI - Twists and turns on G-protein signalling pathways. PMID- 15336045 TI - Mapping the brain with MRI. PMID- 15336046 TI - Evolutionary junk or first class tag-along? PMID- 15336047 TI - Re-emergence of the captain of the men of death. PMID- 15336049 TI - Cloners quick on the uptake. PMID- 15336048 TI - Signals from focal adhesions. PMID- 15336050 TI - Molecular matchmakers. PMID- 15336051 TI - Opening a window on immunosuppression. PMID- 15336052 TI - Can genes be truly redundant? PMID- 15336053 TI - Parallel pathways converge. PMID- 15336054 TI - Light chain surrogacy. PMID- 15336055 TI - Fishing deep beneath the shallow water. PMID- 15336057 TI - The worm turns to vinculin function. PMID- 15336056 TI - Complexes with a common core. PMID- 15336058 TI - jockey leads by a short head. PMID- 15336059 TI - Alzheimer's disease - reassessing the data. PMID- 15336060 TI - How to zap a kappa. PMID- 15336061 TI - On the evolutionary origin of right handedness. PMID- 15336062 TI - Immunosuppressants hit the target. PMID- 15336063 TI - Fine focus on glutamate receptors. PMID- 15336064 TI - It is easier for a camel to pass the needle's eye. PMID- 15336065 TI - Having a blast with gene transfer. PMID- 15336067 TI - II for T, and T4 too. PMID- 15336066 TI - MyoD and the paradoxes of myogenesis. PMID- 15336068 TI - Finding filament function. PMID- 15336069 TI - Gene translocation between organelles. PMID- 15336071 TI - Kinked, curved or bent but certainly not going straight. PMID- 15336070 TI - Tip links and hair cells. PMID- 15336072 TI - Elementary science education in the United States: how scientists can help. PMID- 15336073 TI - Pax in our time. PMID- 15336075 TI - A giant step for myosin. PMID- 15336074 TI - Breaking the topological dogma. PMID- 15336076 TI - Mixing and matching calcium channels. PMID- 15336077 TI - Plugging the GAPs. PMID- 15336078 TI - Of kit and mouse and man. PMID- 15336079 TI - Heat shock proteins and autoimmunity: facts or fiction? PMID- 15336080 TI - Population explosion in the cyclin family. PMID- 15336081 TI - Activating chromatin. PMID- 15336082 TI - Bristling with teeth. PMID- 15336083 TI - Number 1 and counting. PMID- 15336084 TI - Prohormone convertases revealed at last. PMID- 15336085 TI - First, trap your enhancer. PMID- 15336086 TI - A nod of recognition. PMID- 15336088 TI - Death from natural and unnatural causes. PMID- 15336087 TI - Second class distinction. PMID- 15336089 TI - Motion perception and the mind-body problem. PMID- 15336090 TI - How do integrins integrate? PMID- 15336091 TI - Turns in the tail. PMID- 15336092 TI - Colon cancer culprit cloned. PMID- 15336093 TI - Two lobes and a cleft. PMID- 15336094 TI - Current biology. PMID- 15336095 TI - Sweeping visions. PMID- 15336096 TI - Who's zooming who? PMID- 15336098 TI - Threads between useful and useless. PMID- 15336097 TI - Cycles within cycles. PMID- 15336099 TI - Viruses getting their heads together. PMID- 15336100 TI - CD5 B cells: a separate lineage at last? PMID- 15336102 TI - Calcium waves and spirals. PMID- 15336101 TI - Fragmenting the fragile X. PMID- 15336103 TI - Is movement perception color blind? PMID- 15336104 TI - Scratching the surface. PMID- 15336105 TI - Hox genes coming to a head. PMID- 15336107 TI - Bind or bleed. PMID- 15336106 TI - Is thymosin-beta4 the missing link? PMID- 15336108 TI - Familial Alzheimer's disease. PMID- 15336109 TI - B-cell superantigens? PMID- 15336110 TI - Saps, hydrophobins and aerial growth. PMID- 15336111 TI - Reactin' with activin. PMID- 15336112 TI - The destiny of reverse transcriptase anatomy. PMID- 15336113 TI - Plateaus in time. PMID- 15336114 TI - Peptides feeling groovy. PMID- 15336116 TI - MAP kinase activation. PMID- 15336115 TI - Introns in pieces. PMID- 15336117 TI - Distinguishing features. PMID- 15336118 TI - Shared partners in receptors. PMID- 15336119 TI - Making scents of olfactory transduction. PMID- 15336120 TI - Deconstructing protein structure. PMID- 15336121 TI - Ghetto legacy. PMID- 15336122 TI - Shuttling proteins go both ways. PMID- 15336124 TI - Characterizing protein folding intermediates. PMID- 15336123 TI - Decoding the retinal colour signal. PMID- 15336125 TI - Vaccination and the immune response. PMID- 15336126 TI - Complex inhibitions. PMID- 15336128 TI - Recombination of motor pattern generators. PMID- 15336127 TI - Auxin receptors take shape. PMID- 15336129 TI - Exploring the catalytic activity of immunophilins. PMID- 15336130 TI - Adherin' with cadherin. PMID- 15336132 TI - Killing comes naturally. PMID- 15336131 TI - To thine own self be true... PMID- 15336133 TI - RNA bent for recognition. PMID- 15336134 TI - Lonesome receptors find their mates. PMID- 15336136 TI - At the heart of the organizing center. PMID- 15336135 TI - Seeing the whole picture. PMID- 15336137 TI - Scratching the back of the cortex. PMID- 15336138 TI - Plague, pox and tyrosine dephosphorylation. PMID- 15336139 TI - To T or not to T: is it selection? PMID- 15336141 TI - Putting the finger on DNA. PMID- 15336140 TI - Rounding up acetylcholine receptors. PMID- 15336142 TI - Membrane glycoprotein choreography. PMID- 15336143 TI - Making science on a volcano. PMID- 15336144 TI - Sevenless and its bride: a marriage of molecules? PMID- 15336145 TI - Cofactor vanishing act. PMID- 15336146 TI - Seeing texture. PMID- 15336147 TI - Death by a thousand cuts. PMID- 15336149 TI - The origins of introns. PMID- 15336148 TI - Demystification of the alloresponse. PMID- 15336150 TI - New pieces in the U5 puzzle. PMID- 15336151 TI - Bad channel genes and weak muscles. PMID- 15336153 TI - Development of left and right - the worm turns. PMID- 15336152 TI - Balancing effector outputs. PMID- 15336154 TI - PDGF in neurons. PMID- 15336155 TI - The wound response of plants. PMID- 15336157 TI - To bend or...? PMID- 15336156 TI - X marks the spot. PMID- 15336158 TI - Computational representations of sonar images in bats. PMID- 15336159 TI - Chaperoning protein repair. PMID- 15336161 TI - New herpesviruses and their relevance for man. PMID- 15336160 TI - Can clonal anergy resist an isotype switch? PMID- 15336162 TI - The center cannot hold. PMID- 15336163 TI - Acid connections. PMID- 15336165 TI - Understanding MCAD deficiency: one cause of cot death. PMID- 15336164 TI - Jumping genes. PMID- 15336166 TI - Regulation of B-cell development by pre-B-cell receptors. PMID- 15336167 TI - Games that herpes viruses play. PMID- 15336168 TI - Protein carpentry. PMID- 15336169 TI - Infectious origins of superantigens. PMID- 15336170 TI - Motor vesicles. PMID- 15336171 TI - The molecular pathology of cystic fibrosis. PMID- 15336172 TI - How complex is the Golgi? PMID- 15336173 TI - Immunoinhibitory genes. PMID- 15336174 TI - Cycling transcripts and the circadian clock. PMID- 15336175 TI - Putting the G in development. PMID- 15336176 TI - Oncogenes, mitochondria and immortality. PMID- 15336177 TI - Neural coding of local and global motion. PMID- 15336178 TI - Understanding how transcriptional enhancers work. PMID- 15336179 TI - Dangerous liaisons. PMID- 15336181 TI - Circuit dynamics and the strength of synapses. PMID- 15336180 TI - Powering the cell cycle. PMID- 15336182 TI - Pas de deux. PMID- 15336183 TI - Protein prenylation: a modification that sticks. PMID- 15336185 TI - Many cell types specified by Notch function. PMID- 15336184 TI - A helping hand from proteoglycans. PMID- 15336186 TI - Teaching an old oncogene new mycs. PMID- 15336187 TI - Sweetening the selectin pot. PMID- 15336188 TI - Current biology. PMID- 15336189 TI - DNA recombination in the brain? PMID- 15336190 TI - More than just scaffolding... PMID- 15336191 TI - Characterizing intermediates in protein folding. PMID- 15336192 TI - Imprinting and the very young. PMID- 15336193 TI - Unravelling immunoglobulin expression. PMID- 15336194 TI - Protein-serine kinase receptors? PMID- 15336196 TI - New recruit to the magnificent seven. PMID- 15336195 TI - Direct interaction or remote control? PMID- 15336197 TI - Cyclins and cell cycle control. PMID- 15336198 TI - Capturing antigen receptor components. PMID- 15336199 TI - Separating the wheat from the chaff. PMID- 15336200 TI - Self-control of self-assembly. PMID- 15336201 TI - From flies to mice and back again. PMID- 15336202 TI - Probing cytoskeletal compartments. PMID- 15336203 TI - Synaptic mechanisms for long-term depression. PMID- 15336205 TI - A protein translocation machine in yeast. PMID- 15336204 TI - Dropping anchor with the lipophosphoglycans. PMID- 15336206 TI - Miocene DNA sequences - a dream come true? PMID- 15336207 TI - Winning by knock-out or just on points? PMID- 15336208 TI - RNA 'tetraloops': living in syn. PMID- 15336209 TI - In vitro replication of plant viral RNA. PMID- 15336210 TI - Bridging the gaps. PMID- 15336211 TI - Pumping peptides. PMID- 15336213 TI - DAM fine progress but a long way to go. PMID- 15336212 TI - Ante-antibody immunity. PMID- 15336214 TI - If the cap fits... PMID- 15336215 TI - Designing the enzyme of the nineties. PMID- 15336216 TI - Where has all the science gone? PMID- 15336217 TI - Leptospirosis. PMID- 15336219 TI - Therapeutic use of bacteriophages. PMID- 15336221 TI - West Nile virus: where are we now? AB - Since the publication of a comprehensive review on West Nile virus (WNV) in 2002, there has been substantial progress in understanding of transmission, epidemiology, and geographic distribution of the virus and manifestations of disease produced by the infection. There have also been advances in development of diagnostic and therapeutic agents and vaccines. Nevertheless, many questions about the epidemic remain unanswered, and several new issues have arisen--for example: whether the epidemic will increase as the virus spreads to the Pacific coast of North America; whether arthropods other than mosquitoes will act as vectors for the infection; whether WNV will spread to South America and cause an epidemic there; whether the distribution of WNV in Asia and Europe will increase; and whether adaptation of WNV to new ecosystems will produce viruses with altered genetic and phenotypic properties. This review aims to provide an update on knowledge of WNV biology that can be used to highlight the advances in the field during the past 2 years and help to define the questions that academic, industrial, and public-health communities must address in development of measures to control WNV disease. PMID- 15336222 TI - A young man with rectal polyps. PMID- 15336223 TI - Mycobacterium avium complex in patients with HIV infection in the era of highly active antiretroviral therapy. AB - Disseminated Mycobacterium avium complex (MAC) infection is a common complication of late-stage HIV-1 infection. Since the advent of highly active antiretroviral therapy (HAART), the rate of MAC infection has declined substantially, but patients with low CD4 cell counts remain at risk. Among patients in the Johns Hopkins cohort with advanced HIV disease, the proportion developing MAC has fallen from 16% before 1996 to 4% after 1996, with a current rate of less than 1% per year. Factors associated with developing MAC include younger age, no use of HAART, and enrollment before 1996. Prophylaxis with azithromycin or clarithromycin is recommended for all patients with CD4 counts less than 50 cells/mL. Optimum treatment for disseminated MAC includes clarithromycin and ethambutol, and another investigation suggests that the addition of rifabutin might reduce mortality. Both prophylaxis and treatment of disseminated MAC can be discontinued in patients who have responded to HAART, and specific guidelines for withdrawing treatment have been published. Although HAART has altered the frequency and outcome of MAC infection, it remains an important complication of AIDS. PMID- 15336224 TI - Mycetoma caused by Madurella mycetomatis: a neglected infectious burden. AB - Tropical eumycetoma is frequently caused by the fungus Madurella mycetomatis. The disease is characterised by extensive subcutaneous masses, usually with sinuses draining pus, blood, and fungal grains. The disease affects individuals of all ages, although disability is most severe in adults who work outdoors. Compared with major diseases such as tuberculosis, malaria, and HIV, disease from M mycetomatis is underestimated but socioeconomically important. Many scientific case reports on mycetoma exist, but fundamental research was lacking until recently. We present a review on developments in the clinical, epidemiological, and diagnostic management of M mycetomatis eumycetoma. We describe newly developed molecular diagnostic and gene typing procedures, and their application for management of patients and environmental research. Fungal susceptibility tests have been developed as well as a mouse model of infection. These advances should greatly further our understanding of the molecular basis of eumycetoma. PMID- 15336225 TI - Molecular survival strategies of the Lyme disease spirochete Borrelia burgdorferi. AB - Lyme disease is a tick-transmitted disease caused by the spirochete Borrelia burgdorferi. The bacterium adopts different strategies for its survival inside the immunocompetent host from the time of infection until dissemination in different parts of body tissues. The success of this spirochete depends on its ability to colonise the host tissues and counteract the host's defence mechanisms. During this process borrelia seems to maintain its vitality to ensure long-term survival in the host. Borrelia's proteins are encoded by plasmid and chromosomal genes. These genes are differentially regulated and expressed by different environmental factors in ticks as well as in the mammalian host during infection. In addition, antigenic diversity enables the spirochete to escape host defence mechanisms and maintain infection. In this review we focus on the differential expression of proteins and genes, and further molecular mechanisms used by borrelia to maintain its survival in the host. In light of these pathogenetic mechanisms, further studies on spirochete host interaction are needed to understand the complex interplay that finally lead to host autoimmunity. PMID- 15336226 TI - Tuberculosis: from prehistory to Robert Koch, as revealed by ancient DNA. AB - During the past 10 years palaeomicrobiology, a new scientific discipline, has developed. The study of ancient pathogens by direct detection of their DNA has answered several historical questions and shown changes to pathogens over time. However, ancient DNA (aDNA) continues to be controversial and great care is needed to provide valid data. Here we review the most successful application of the technology, which is the study of tuberculosis. This has provided direct support for the current theory of Mycobacterium tuberculosis evolution, and suggests areas of investigation for the interaction of M tuberculosis with its host. PMID- 15336227 TI - Pushing Bordeaux mixture. PMID- 15336229 TI - Free insulin-like growth factors -- measurements and relationships to growth hormone secretion and glucose homeostasis. AB - IGF-I is a multipotent growth factor with important actions on normal tissue growth and regeneration. In addition, IGF-I has been suggested to have beneficial effects on glucose homeostasis due to its glucose lowering and insulin sensitizing actions. However, not all effects of IGF-I are considered to be favorable; thus, epidemiological studies suggest that IGF-I is also involved in the development of common cancers, atherosclerosis and type 2 diabetes. The biological actions of IGF-I are modulated by at least six IGF-binding proteins, which bind approximately 99% of the circulating IGF-I pool. So far, most in vivo studies have used serum or plasma total (extractable IGF-I) as an estimate of the bioactivity of IGF-I in vivo. However, within the last decade, validated assays for measurement of free IGF-I have been described. This review aims to discuss the current assays for free IGF-I and their advances in relation to the traditional measurement of total IGF-I. The literature overview will focus on the role of circulating free versus total IGF-I in the feedback regulation of GH release, and the possible involvement of the circulating IGF-system in glucose homeostasis. PMID- 15336228 TI - What's new in the IGF-binding proteins? AB - Since their initial discovery over 25 years ago as IGF carrier proteins, the insulin-like growth factor binding protein (IGFBP) family has grown to six members, ranging in size from 216 to 289 amino acids. The assumption over the years has been that this family of proteins, having higher affinities for IGF-I and IGF-II than does the IGF-IR, serves to block access of these ligands to the receptor. Although the need for such regulatory proteins is consistent with the constitutive secretion of IGFs from many cell types, it is not surprising that additional functions have begun to be uncovered for these proteins. This review will examine new and old actions of the IGFBPs from a biochemical and cell biological perspective. PMID- 15336230 TI - The effect of insulin-like growth factor-I on production of vascular endothelial growth factor by amnion-derived (WISH) cells. AB - OBJECTIVE: Our objective was to clarify the modulation of vascular endothelial growth factor (VEGF) by amniotic cells. DESIGN: Amnion-derived (WISH) cells were cultured, and the effect of insulin-like growth factor (IGF), mitogen-activated protein (MAP) kinase kinase and/or extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitors (U0126), and phosphatidylinositol (PI) 3-kinase inhibitors (wortmannin) on the production of VEGF was examined. VEGF was assayed using ELISA. The activations of MAP kinase and akt, which is phosphorylated by PI 3-kinase, were detected by Western blot analysis using anti-phosphorylated MAP kinase antibody and anti-phosphorylated akt antibody. RESULTS: In the time course of VEGF production following IGF-I treatment, VEGF production showed significant increases only at 16 and 32 h (p < 0.01). Also, IGF-I increased the production of VEGF by WISH cells in a dose-dependent manner. The MAP kinase and akt activities were recognized by treatment with IGF-I and suppressed by U0126 and wortmannin, respectively. When WISH cells were pretreated for 2 h with U0126 and wortmannin and then treated with IGF-I for 16 h, the production of VEGF was significantly decreased in a dose-dependent manner (p < 0.01, p < 0.01, respectively). CONCLUSIONS: WISH cells appeared to produce VEGF via a mechanism involving tyrosine kinase interaction with IGF-I receptor, resulting in MAP kinase and PI 3 kinase activation. It is suggested that VEGF may contribute to the neovascularization and proliferation of the placenta and gestational tissue, and IGF-I may play an important role in the modulation of VEGF production in the placenta. PMID- 15336231 TI - Cortistatin-17 and -14 exert the same endocrine activities as somatostatin in humans. AB - Cortistatin (CST) is a neuropeptide, which binds with high affinity all somatostatin (SS) receptor subtypes and shows high structural homology with SS itself. A receptor specific for CST only, i.e., not recognized by SS, has been recently described in agreement with data reporting that not all CST actions are shared by SS. Interestingly, CST but not SS also binds ghrelin receptor (GHS-R1a) in vitro, suggesting a potential interplay between CST and ghrelin system. The aim of this study was to investigate in humans the endocrine and metabolic activities of human CST-17 in comparison with rat CST-14 that has previously been shown to exert the same endocrine actions of SS in healthy volunteers. To this aim, in six healthy male volunteers (age [median, 3rd-97th centiles]: 28.5; 23.6 34.3 years; Body Mass Index: 23.5; 21.0-25.1 kg/m(2)), we studied the effects of human CST-17 (2.0 microg/kg/h iv over 120 min), rat CST-14 (2.0 microg/kg/h iv over 120 min) and SS-14 (2.0 microg/kg/h iv over 120 min) on: (a) spontaneous GH, ACTH, PRL, cortisol, insulin and glucose levels; (b) the GH responses to GHRH (1.0 microg/kg iv at 0 min); (c) the GH, PRL, ACTH, cortisol, insulin and glucose responses to ghrelin (1.0 microg/kg iv at 0 min). CST-17 inhibited (p < 0.01) basal GH secretion to the same extent of CST-14 and SS-14. Spontaneous PRL, ACTH and cortisol secretion were not significantly modified by CST-17, CST-14 or SS 14. CST-17 as well as CST-14 and SS-14 also inhibited (p < 0.05) spontaneous insulin secretion to a similar extent. None of these peptides modified glucose levels. The GH response to GHRH was inhibited to the same extent by CST-17 (p < 0.01), CST-14 (p < 0.01) and SS-14 (p < 0.05 ). The ghrelin-induced GH response was higher than that elicited by GHRH (p < 0.01) and inhibited by CST-17 (p < 0.05) as well as by CST-14 (p < 0.05) and SS-14 (p < 0.01). The PRL, ACTH and cortisol responses to ghrelin were unaffected by CST-17, CST-14 or SS-14. On the other hand, the inhibitory effect of ghrelin on insulin levels was abolished by CST-17, CST-14 or SS-14 (p < 0.05) that, in turn, did not modify the ghrelin induced increase in glucose levels. In conclusion, this study demonstrates that human CST-17 and rat CST-14 exert the same endocrine activities of SS in humans. The endocrine actions of human and rat CST therefore are likely to reflect activation of classical SS receptors. PMID- 15336232 TI - Ovariectomy and gonadal hormone treatment: effects on insulin-like growth factor 1 receptors in the rat brain. AB - Various studies demonstrate that estradiol regulates structure and function of adult neurons. Long-term effect of estradiol in terms of neuroprotection is less documented compared to short-term one. It is well documented that estradiol interacts with insulin-like growth factor-1 (IGF-I) in the brain. The present study examines the effect of ovariectomy and two doses of ovarian hormone treatment on IGF-I receptor density in the adult rat by receptor autoradiography using (125)I-IGF-I as a ligand. Our result showed that ovariectomy decreased IGF I receptor density in hippocampus, hypothalamus and parietal cortex compared to that of the sham-operated group. Treatment with low or high dose estrogen restored IGF-I receptor density to the control levels in nearly all areas studied in this investigation. It seems that low dose estrogen has more pronounced effect than the high dose in restoring IGF-I receptor density. On the other hand, progesterone treatment in high but not in low dose restored IGF-I receptor density to that of the control. These results demonstrate that both estrogen and progesterone significantly affects IGF-I receptor density in different areas of the brain. These effects indicate a dose-dependent modulator effect of ovarian hormones on IGF-I activity in the brain. PMID- 15336233 TI - A nonsense mutation (E72X) in growth hormone releasing hormone receptor (GHRHR) gene is the major cause of familial isolated growth hormone deficiency in Western region of India: founder effect suggested by analysis of dinucleotide repeat polymorphism close to GHRHR gene. AB - An identical nonsense mutation (E72X) in growth hormone releasing hormone receptor (GHRHR) gene was identified in 17 patients with isolated GH deficiency belonging to one Muslim and four Hindu families residing in the Western part of India. Analysis of two dinucleotide repeat polymorphism, one at 6 kb downstream and the other at 13 kb downstream of GHRHR gene, revealed that all the patients shared the same homozygotic alleles at both loci. These results strongly indicate that the nonsense mutation occurred in a single ancestor and was subsequently transmitted to the descendants. This GHRHR mutation may be an important cause of familial IGHD in Western India and Sindh area of Pakistan as previous studies have also reported the same mutation. PMID- 15336234 TI - An update on the pharmacotherapy of excessive daytime sleepiness and cataplexy. PMID- 15336235 TI - Pharmacotherapy for excessive daytime sleepiness. AB - Excessive daytime sleepiness (EDS) has recognized detrimental consequences such as road traffic accidents, impaired psychological functioning and reduced work performance. EDS can result from multiple causes such as sleep deprivation, sleep fragmentation, neurological, psychiatric and circadian rhythm disorders. Treating the underlying cause of EDS remains the mainstay of therapy but in those who continue to be excessively sleepy, further treatment may be warranted. Traditionally, the amphetamine derivatives, methylphenidate and pemoline (collectively sympathomimetic) psychostimulants were the commonest form of therapy for EDS, particularly in conditions such as narcolepsy. More recently, the advent of modafinil has broadened the range of therapeutic options. Modafinil has a safer side-effect profile and as a result, interest in this drug for the management of EDS in other disorders, as well as narcolepsy, has increased considerably. There is a growing school of thought that modafinil may have a role to play in other indications such as obstructive sleep apnea/hypopnea syndrome already treated by nasal continuous positive airway pressure but persisting EDS, shift work sleep disorders, neurological causes of sleepiness, and healthy adults performing sustained operations, particularly those in the military. However, until adequately powered randomised-controlled trials confirm long-term efficacy and safety, the recommendation of wakefulness promoters in healthy adults cannot be justified. PMID- 15336236 TI - Pharmacotherapy for cataplexy. AB - A variety of medications representing several major drug classes improve cataplexy in patients with narcolepsy. These include aminergic reuptake inhibitors such as venlafaxine and clomipramine as well as sodium oxybate. This review is intended to familiarize readers with the safety and efficacy of these medications, thus enabling clinicians to optimize their management of cataplexy. PMID- 15336237 TI - GHRH and sleep. AB - A significant portion of the total daily growth hormone (GH) secretion is associated with deep non-REM sleep (NREMS). GH secretion is stimulated by the hypothalamic neurohormone, GH-releasing hormone (GHRH). Exogenous GHRH promotes NREMS in various species. Suppression of endogenous GHRH (competitive antagonist, antibodies, somatostatinergic stimulation, high doses of GH or insulin-like growth factor) results in simultaneous inhibition of NREMS. Mutant and transgenic animals with a defect in GHRHergic activity display permanently reduced NREMS which cannot be reversed by means of GH supplementation. GHRH contents and mRNA levels in the hypothalamus correlate with sleep-wake activity during the diurnal cycle and sleep deprivation and recovery sleep. Stimulation of NREMS by GHRH is a hypothalamic action. GABAergic neurons in the anterior hypothalamus/preoptic region are candidates for mediating promotion of NREMS by GHRH. In contrast to NREMS, stimulation of REMS by GHRH is mediated by GH. Simultaneous stimulation of NREMS and GH secretion by GHRH may promote adjustment of tissue anabolism to sleep. PMID- 15336238 TI - Sleep in children with attention-deficit hyperactivity disorder (ADHD): a review of naturalistic and stimulant intervention studies. AB - Attention-Deficit Hyperactivity Disorder (ADHD) is the most common behavioral disorder of childhood. Multiple clinical and research reports suggest extensive sleep disturbances in children with ADHD, however, current data is contradictory. This paper reviewed 47 research studies (13 stimulant intervention and 34 naturalistic) on ADHD that were published since 1980. The main objectives of this review were to provide pediatric clinicians and researchers a clear and concise summary of published sleep data in children with ADHD, to provide a more accurate description of the current knowledge of the relationship between sleep and ADHD, and to provide current information on the effect of stimulant medication on sleep. Twenty-five of the reviewed studies used subjective reports of sleep, six were actigraphic studies, and 16 were overnight polysomnographic sleep studies (two of which also included Multiple Sleep Latency Tests). All participants were between the age of 3 and 19, and 60% were male. The results indicate high rates of parental reports of sleep disturbances in medicated and unmedicated children with ADHD, however, the majority of these findings have not been confirmed by objective sleep data. Although, agreement among objective studies is not absolute, the data suggest increased nighttime activity, reduced rapid eye movement sleep, and significant daytime somnolence in unmedicated children with ADHD when compared to controls. Data also suggest a possible increased prevalence of periodic limb movements in sleep in children with ADHD, however, little differences in sleep-disordered breathing. The limited number of studies, small and heterogeneous samples, and other methodological limitations make definite results difficult to determine. Future research will need to further clarify the relationship between sleep and ADHD and the effects of stimulants on sleep of children with ADHD. PMID- 15336239 TI - Chronobiological features of dream production. AB - A review of the scientific literature clarifies several chronobiological features of dreaming. The literature supports the conclusions that dreaming 'intensity' and, to a lesser extent dream-like quality, is modulated by (1) a sinusoidal, 90 min ultradian oscillation, (2) a 'switch-like' circadian oscillation, (3) a 12-h circasemidian rhythm, and (4) a 28-day circatrigintan rhythm (for women). Further, access to dream memory sources appears to be modulated by (5) a 7-day circaseptan rhythm. Further study of these rhythmic influences on dreaming may help to explain diverse and often contradictory findings in the dream research literature, to clarify relationships between dreaming and waking cognitive processes, to explain relationships between disturbed phase relationships and dream disturbances and to shed new light on the problems of dreaming's functions and biological markers. Further chronobiological studies of dreaming will likely enable the development of theoretical models that explain how interactions between and within major levels of oscillation determine the variable characteristics of dreaming. PMID- 15336240 TI - Contingency awareness in a symptom learning paradigm: necessary but not sufficient? AB - In previous studies, we found that bodily symptoms can be learned in a differential conditioning paradigm, using odors as conditioned stimuli (CSs) and CO2-enriched air as unconditioned stimulus (US). However, this only occurred when the odor CS had a negative valence (a selective conditioning effect), and tended to be more pronounced in persons scoring high for Negative Affectivity (NA). This paper considers the necessity and/or sufficiency of awareness of the CS-US contingency in three studies using this paradigm. The relation between contingency awareness and the selective conditioning effect, and between contingency awareness and NA was also considered. Both self reported symptoms and respiratory physiology served as dependent variables. A learning effect on symptoms was found only for participants aware of the CS-US contingency, but not all participants reporting contingency awareness showed a learning effect. No conditioning effects appeared on the physiological measures. Also contingency awareness did not account for the selective conditioning effect, and did not interact with NA. Overall, the necessity but insufficiency hypotheses can only be withhold for group data and not for individual data. PMID- 15336241 TI - Does dietary learning occur outside awareness? AB - Several forms of dietary learning have been identified in humans. These include flavor-flavor learning, flavor-postingestive learning (including flavor-caffeine learning), and learned satiety. Generally, learning is thought to occur in the absence of contingency (CS-US) or demand awareness. However, a review of the literature suggests that this conclusion may be premature because measures of awareness lack the rigor that is found in studies of other kinds of human learning. If associations do configure outside awareness then this should be regarded as a rare instance of automatic learning. Conversely, if awareness is important, then successful learning may be governed by an individual's beliefs and predilection to attend to stimulus relationships. For researchers of dietary learning this could be critical because it might explain why learning paradigms have a reputation for being unreliable. Since most food preferences are learned, asking questions about awareness can also tell us something fundamental about everyday dietary control. PMID- 15336242 TI - Implicit contributions of context to recognition. AB - Studies of the impact of context on remembering have not focused on the influence of contextual contingency on subsequent recognition in the condition in which the contingency cannot be verbalized. In two experiments, we analyzed the effect of an implicitly encoded position contingency involving location and semantic category on both hit and false alarm recognition judgments after 1 day and 1 week delays. We vigorously probed for what participants could say about the contingency. We found context effects for both hits and false alarms, whether or not participants could verbalize any knowledge they might have of contingency. These results suggest that people may use contextual information when making a recognition judgment even if they are not aware of this information. PMID- 15336243 TI - Poor recall of eye-movement signals from Stage 2 compared to REM sleep: implications for models of dreaming. AB - An ongoing assumption made by sleep researchers is that since dreams are more often recalled on awakening from rapid eye movement (REM) sleep, dreams must occur more often during this stage of sleep. An alternative hypothesis is that cognition occurs throughout sleep, but the recall of this mentation differs on awakening. When a dream is not reported on awakening, there is no method of establishing whether it did not happen or was forgotten. The aim of the present study was to investigate this issue using an eye movement (EM) signal verification technique. Participants were instructed to produce an EM signal whenever they heard a tone. Tones were presented at increasing volume during Stage 2 and REM sleep until EM signal verification was observed. Ninety seconds after signal verification, participants were awakened and asked if they remembered hearing the tone or responding with the EM signal. Such recollection of signal verified tone (SVT) presentations was significantly less after Stage 2 sleep (65%) compared to REM sleep (100%) presentations. Furthermore, SVT recall was significantly correlated with reported dream recall frequency, suggesting the same processes involved in recalling SVTs might also underlie dream recall. PMID- 15336244 TI - Motivation and affect in REM sleep and the mentation reporting process. AB - Although the emotional and motivational characteristics of dreaming have figured prominently in folk and psychoanalytic conceptions of dream production, emotions have rarely been systematically studied, and motivation, never. Because emotions during sleep lack the somatic components of waking emotions, and they change as the sleeper awakens, their properties are difficult to assess. Recent evidence of limbic system activation during REM sleep suggests a basis in brain architecture for the interaction of motivational and cognitive properties in dreaming. Motivational and emotional content in REM and NREM laboratory mentation reports from 25 participants were compared. Motivational and emotional content was significantly greater in REM than NREM sleep, even after controlling for the greater word count of REM reports. PMID- 15336245 TI - Is unconscious identity priming lexical or sublexical? AB - We examined unconscious priming in a stem-completion task with both identity and form-related primes. Participants were given exclusion instructions to avoid completing a stem (e.g., ca---) with a briefly flashed masked word (e.g., candy). In Experiment 1, priming of around 7% occurred for both identity (e.g., candy) and form-based (e.g., windy) primes at a 33 ms exposure duration. When examining only trials in which the participants failed to identify the prime, this effect increased to 12% for identity primes, but remained the same for form-based primes. In Experiment 2, priming without prime identification was 9% for identity primes, 4% for homophone primes, and 3% for orthographic control primes. Although identity priming was greater than form priming in both experiments, regression analyses revealed that orthographic and phonological overlap alone between the flashed primes and targets could completely account for unconscious identity priming. Hence, we conclude that masked words may only activate their sublexical orthographic and phonological representations and not their lexical representations. PMID- 15336246 TI - Extending the global workspace theory to emotion: phenomenality without access. AB - Recent accounts on the global workspace theory suggest that consciousness involves transient formations of functional connections in thalamo-cortico cortical networks. The level of connectivity in these networks is argued to determine the state of consciousness. Emotions are suggested to play a role in shaping consciousness, but their involvement in the global workspace theory remains elusive. In the present study, the role of emotion in the neural workspace theory of consciousness was scrutinized by investigating, whether unconscious (masked) and conscious (unmasked) display of emotional compared to neutral facial expressions would differentially modulate EEG coherence. EEG coherence was measured by means of computing an average EEG coherence value between the frontal, parietal, and midline scalp sites. Objective awareness checks evidenced that conscious identification of the masked facial expressions was precluded. Analyses revealed reductions in EEG coherence in the lower frequency range for the masked as compared to unmasked neutral facial expressions. Crucially, a decline in EEG coherence was not observed for the emotional facial expressions. In other words, the level of EEG coherence did apparently vary as a function of awareness, but not when emotion was involved. The current finding suggests that EEG coherence is modulated by unconscious emotional processes, which extends common views on the global workspace architecture of consciousness. PMID- 15336247 TI - Hemisphere differences in conscious and unconscious word reading. AB - Hemisphere differences in word reading were examined using explicit and implicit processing measures. In an inclusion task, which indexes both conscious (explicit) and unconscious (implicit) word reading processes, participants were briefly presented with a word in either the right or the left visual field and were asked to use this word to complete a three-letter word stem. In an exclusion task, which estimates unconscious word reading, participants completed the word stem with any word other than the prime word. Experiment 1 showed that words presented to either visual field were processed in very similar ways in both tasks, with the exception that words in the right visual field (left hemisphere) were more readily accessible for conscious report. Experiment 2 indicated that unconsciously processed words are shared between the hemispheres, as similar results were obtained when either the same or the opposite visual field received the word stem. Experiment 3 demonstrated that this sharing between hemispheres is cortically mediated by testing a split-brain patient. These results suggest that the left hemisphere advantage for word reading holds only for explicit measures; unconscious word reading is much more balanced between the hemispheres. PMID- 15336248 TI - Opposition logic and neural network models in artificial grammar learning. AB - Following neural network simulations of the two experiments of, argued that the opposition logic advocated by was incapable of distinguishing between single and multiple influences on performance of artificial grammar learning and more generally. We show that their simulations do not support their conclusions. We also provide different neural network simulations that do simulate the essential results of Higham et al. (2000). PMID- 15336249 TI - A neuroanatomical model of passivity phenomena. AB - Any attempt to elucidate the nature and mechanism of passivity phenomena, i.e., experiences that one's conscious actions or thoughts have not been 'willed' by oneself, requires an integrative philosophical-neurobiological approach. The model proposed here adopts some fundamental positions that have long been advocated by philosophers and theoretical psychologists and have now found support from functional neuroanatomy. First, we experience our actions not from the standpoint of the executive but through the perception of its effects. Second, the 'self' is not an agent of behaviour. Third, behaviour is energized and integrated by basic drives (instincts). Fourth, the view that the experience of an acting self is related to drive reduction associated with voluntary actions is perhaps less well developed. The model thus proposes that passivity phenomena are actions that are induced by the perception of salient events but that are not integrated with or conducive to the overall motivational state of the organism. It has been suggested that, following the perception of salient events, competition arises between automatic response tendencies seeking expression. The prefrontal cortex appears to play an important role not only in determining which events are to be perceived but also which of the corresponding response dispositions is to be selected and actualized in overt behaviour. Thus, action selection is the outcome of competition between response tendencies in the context of prefrontal biasing signals that represent drives and strivings for goals. Action selection may be uncoupled from drives and strivings as a result of a lowering of the threshold for action selection--as is suggested to be the case in schizophrenic passivity phenomena--or due to disconnection from prefrontal regions--as may be the case in the alien limb syndrome. PMID- 15336250 TI - Cross-modal facilitation is not specific to self-face recognition. PMID- 15336251 TI - Subliminality, consciousness, and temporal shifts in awareness: implications within and beyond the laboratory. AB - In his analysis of subliminal perception research, documented two important (but neglected) phenomena: subchance perception and temporal variability in stimulus availability and accessibility. This Commentary addresses three issues raised by Erdelyi's review: (1) the importance of distinguishing "micro" from "macro" temporal shifts; (2) the need to analyze perception without awareness data at the level of the individual as well as the group; and (3) parallels between the dissociations associated with neuroclinical phenomena (e.g., hemispheric isolation) and those observed in patients with certain forms of personality pathology. Continued integration of laboratory findings with in vivo observations of clinical syndromes will yield a more cohesive and heuristic approach to the study of implicit mental states. PMID- 15336252 TI - Hypnosis and the death of "subjective backwards referral". PMID- 15336253 TI - Brain stimulation and conscious experience. AB - Libet discovered that a substantial duration (> 0.5-1.0 s) of direct electrical stimulation of the surface of the somatosensory cortex at threshold currents is required before human subjects can report that a conscious somatosensory experience had occurred. Using a reaction time method we confirm that a similarly long stimulation duration at threshold currents is required for activation of elementary visual experiences (phosphenes) in human subjects following stimulation of the surface of the striate cortex. However, the reaction times for the subject to respond to the cessation of the visual experience after the end of electrical stimulation could be as brief as 225-242 ms. We also carried out extensive studies in cats under a variety of anesthetic conditions using the same electrodes and parameters of stimulation employed in the human studies to study the patterns of neuronal activity beneath the stimulating surface electrode. Whereas sufficiently strong currents can activate neurons within milliseconds, stimulating currents close to threshold activate sustained neural activity only after at least 350-500 ms. When currents are close to threshold, some neurons are inhibited for several hundreds of millisecond before the balance between inhibition and excitation shifts towards excitation. These results suggest that the prolonged latencies, i.e., latencies beyond 200-250 ms, for the emergence of conscious experience following direct cortical stimulation result from a delay in the sustained activation of underlying cortical neurons at threshold currents rather than being due to any unusually long duration in central processing time. Intracellular records from cortical neurological cells during repetitive electrical stimulation of the surface of the feline striate cortex demonstrate that such stimulation induces a profound depolarizing shift in membrane potential that may persist after each stimulus train. Such a depolarization is evidence that extracellular K+ concentrations have increased during electrical stimulation. Such an increase in extracellular K+ progressively increases cortical excitability until the threshold for sustained activation of cortical neurons is reached and then exceeded. Consequently, the long latency for threshold activation of cortical neurons depends upon a dynamically increasing cortical facilatory process that begins hundreds of milliseconds before there is sustained activation of such neurons. In some cases, this facilatory process must overcome an initial stimulus-induced inhibition before neuronal firing commences. PMID- 15336254 TI - Anomalous control: when 'free-will' is not conscious. AB - The conscious feeling of exercising 'free-will' is fundamental to our sense of self. However, in some psychopathological conditions actions may be experienced as involuntary or unwilled. We have used suggestion in hypnosis to create the experience of involuntariness (anomalous control) in normal participants. We compared a voluntary finger movement, a passive movement and a voluntary movement suggested by hypnosis to be 'involuntary.' Hypnosis itself had no effect on the subjective experience of voluntariness associated with willed movements and passive movements or on time estimations of their occurrence. However, subjective time estimates of a hypnotically-suggested, 'involuntary' finger movement were more similar to those for passive movements than for voluntary movements. The experience of anomalous control is qualitatively and quantitatively different from the normal conscious experience of a similar act produced intentionally. The experience of anomalous control may be produced either by pathology, or, in our case, by suggestion. PMID- 15336255 TI - Synthesis of ethyleneoxide modified 3-carboranyl thymidine analogues and evaluation of their biochemical, physicochemical, and structural properties. AB - Eleven 3-carboranyl thymidine analogues (3CTAs) containing highly hydrophilic and flexible ethyleneoxide moieties were synthesized as potential agents for boron neutron capture therapy (BNCT) and their biochemical and physicochemical properties were evaluated. Based on specific structural features, this library of 3CTAs was divided into three subgroups. The first group contained 3CTAs with 1-4 ethyleneoxide units between the thymidine (Thd) scaffold and a carborane cluster. The second group of 3CTAs contained a pentylene spacer between Thd and the carborane and 2-4 ethyleneoxide units additionally attached to the carborane cluster. The third group contained three 3CTAs all with pentylene spacers and four ethylene units but with different carborane cages. The ethyleneoxide modified 3CTAs were good substrates of thymidine kinase 1 (TK1) and poor substrates of human mitochondrial thymidine kinase 2 (TK2) as determined in phosphoryl transfer assays. In the first group of 3CTAs, all the compounds were efficiently phosphorylated regardless of varying spacer lengths (37-42% of the activity of Thd). The second group of 3CTAs was less effectively phosphorylated (17-26% of the activity of Thd) probably due to a less favorable sterical orientation of Thd within the active site of TK1 and/or an increased lipophilicity compared with the first group. In the third group of structural isomers, no significant differences in phosphorylation rates were observed (17 25%). A structure-function hypothesis explaining these results is presented. PMID- 15336256 TI - Systematic synthesis of four epicatechin series procyanidin trimers and their inhibitory activity on the Maillard reaction and antioxidant activity. AB - A systematic synthesis of four natural epicatechin series procyanidin trimers [[4,8:4",8"]-2,3-cis-3,4-trans: 2",3"-cis-3",4"-trans: 2,3-trans-(-)-epi-catechin (-)-epicatechin-(+)-catechin, [4,8:4",8"]-2,3-cis-3,4-trans: 2",3"-cis-3",4" trans: 2,3-cis-tri-(-)-epicatechin: procyanidin C1, [4,8:4",8"]-2,3-cis-3,4 trans: 2",3"-trans-3",4"-trans: 2,3-trans-(-)-epicatechin-(+)-catechin-(+) catechin: procyanidin C4, and [4,8:4",8"]-2,3-cis-3,4-trans: 2",3"-trans-3",4" trans: 2,3-cis-(-)-epicatechin-(+)-catechin-(-)-epicatechin] is described. Condensation of (2R,3R,4S)-5,7,3'4'-tetra-O-benzyl-4-(2"-ethoxyethyloxy)flavan derived from (-)-epicatechin as an electrophile with the dimeric nucleophiles in the presence of TMSOTf followed by deprotection yielded trimers. Inhibitory activities on the Maillard reaction and antioxidant activity on lipid peroxide of the synthesized oligomers were also investigated. PMID- 15336257 TI - A highly bioactive lignophenol derivative from bamboo lignin exhibits a potent activity to suppress apoptosis induced by oxidative stress in human neuroblastoma SH-SY5Y cells. AB - Approaches to protection against neurodegenerative diseases, in which oxidative stress and inflammation are implicated, should be based on the current concept on the etiology of these diseases. Recently, a new therapeutic strategy has been proposed to protect neurons from cell death by attenuating the apoptotic signal transduction. Lignin, a durable aromatic network polymer second to cellulose in abundance, was able to be converted into highly active lignophenol derivatives with antioxidant activity by using our newly developed phase-separation technique. These lignophenol derivatives were found to show the potent neuroprotective activity against oxidative stress. Among the compounds examined, a lignocresol derivative from bamboo (lig-8) exhibited the most potent neuroprotective activity against hydrogen peroxide (H(2)O(2))-induced apoptosis in human neuroblastoma cell line SH-SY5Y by preventing the caspase-3 activation via either caspase-8 or caspase-9. Furthermore, it was found that lig-8 exerted the antiapoptotic effect by inhibiting dissipation of the mitochondrial membrane permeability transition induced by H(2)O(2) or by the peripheral benzodiazepin receptor ligand PK11195. Lig-8 was also shown to be potent in the antioxidant activity in the cells exposed to H(2)O(2), as assessed by flow cytometry using 5 (and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate and in vitro reactive oxygen species-scavenging potency. These data suggest that lig-8 is a promising neuroprotector, which affects the signaling pathway of neuronal cell death and that it would be of benefit to delay the progress of neurodegenerative diseases. PMID- 15336258 TI - Synthesis, stereochemical determination and biochemical characterization of the enantiomeric phosphate esters of the novel immunosuppressive agent FTY720. AB - The novel immunosuppressive agent FTY720 (1) is phosphorylated in vivo in a variety of species yielding an active metabolite that is an agonist of four of the five known G-protein-coupled sphingosine-1-phosphate (S1P) receptors. A synthesis amenable to producing gram quantities of the stereoisomeric phosphate esters, a determination of their absolute stereochemistry via an enantioselective synthesis and their characterization as S1P receptor agonists and antagonists is reported. PMID- 15336259 TI - Quinones as antimycobacterial agents. AB - Mycobacterium tuberculosis is a serious worldwide health threat, killing almost 3 million people per year. Other mycobacterial species, especially Mycobacterium avium, are emerging pathogens in the immunocompromised population, most notably AIDS patients. These nontuberculous mycobacteria (NTM) are ubiquitous in the environment, and naturally resistant to many disinfection procedures. Treatment options are limited, and no new antibiotics have been developed against mycobacteria since the 1970s. There is a desperate need for new biocides and antibiotics to prevent and treat mycobacterial infections. A small aromatic compound library has been screened for effectiveness in growth inhibition or killing of mycobacteria. Four species, representing the M. tuberculosis complex, the slow-growing NTM, and the rapid-growing NTM were used. Active compounds had minimal inhibitory concentrations as low as 12.5 microg/mL, with the active component being a quinone. The primarily bactericidal activity observed represents a unique mechanism of action. A fluorescent assay involving M. smegmatis expressing gfp was analyzed as a rapid assay for predicting inhibitory activity, but failed to predict activity well. Our compounds may have significant utility as soluble biocides against mycobacteria and other hardy nosocomial pathogens. PMID- 15336260 TI - Stochastic-based descriptors studying peptides biological properties: modeling the bitter tasting threshold of dipeptides. AB - MARCH-INSIDE methodology was applied to the prediction of the bitter tasting threshold of 48 dipeptides by means of pattern recognition techniques, in this case linear discriminant analysis (LDA), and regression methods. The LDA models yielded a percentage of good classification higher than 80% with the two main families of descriptor generated by this methodology (95.8% for self return probability and 83.3% using electronic delocalization entropy). The regression models can explain more than 80% of the experimental variance of the independent variable. Two regression models were obtained with R(2) values of 0.82 and 0.88 for the whole data and the data without two outliers, respectively; having a standard deviation of 0.27 and 0.23. The predictive power of the obtained equations was assessed by the Leave-One-Out cross validation procedures, giving the same percentages of good classification as in the training set, in the LDA models, and yielding values of q(2) of 0.78 and 0.86 in the regression model, respectively. The validation of this methodology was also carried out by comparison with previous reports modeling this data with other well-known methodologies, even 3-D molecular descriptors. PMID- 15336261 TI - New 3-piperonylcoumarins as inhibitors of glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) from Trypanosoma cruzi. AB - This article describes the synthesis and inhibitory activities of a series of new 3-piperonylcoumarins, designed as inhibitors of glycosomal glyceraldehyde-3 phosphate dehydrogenase (gGAPDH) from Trypanosoma cruzi. The design was based on the structures of previously identified natural products hits. The most active synthesized derivatives contain heterocyclic rings at position 6. SAR studies, performed by electronic indices methodology (EIM), clustered the molecules in different groups due to the chemical substitutions regarding the biological activity. Molecular modeling studies by docking suggested a different binding mode for the most active derivatives, when compared to natural hit chalepin. Moreover, the coumarin ring seems to act only as a spacer group. PMID- 15336262 TI - Synthesis of chroman analogues of lipoic acid and evaluation of their activity against reperfusion arrhythmias. AB - Novel hybrids of lipoic acid and trolox connected through triamine spacers as well as analogues in which the lipoic acid was attached at different positions of the chroman moiety of vitamin E through an amide bond, were synthesized and exhibited strong inhibition of the microsomal lipid peroxidation. Moreover, the new molecules, at 1 microM concentration, reduced reperfusion arrhythmias and MDA content on isolated rat heart preparations, with the 2- and 5-subtituted chromans possessing the better cardioprotective activity. PMID- 15336263 TI - Dual 5-HT1A agonists and 5-HT re-uptake inhibitors by combination of indole-butyl amine and chromenonyl-piperazine structural elements in a single molecular entity. AB - The dual serotonin (5-HT) re-uptake inhibitor and 5-HT(1A) receptor agonist vilazodone was found to increase central serotonin levels in rat brain. In the course of structural modifications of vilazodone 3-[4-[4-(2-oxo-2H-1-benzopyran-6 yl)-1-piperazinyl]-butyl]-1H-indole-5-carbonitrile 8i and its fluorine analogue 6 [4-[4-(5-fluor-3-indolyl)-butyl]-1-piperazinyl]-2H-1-benzopyran-2-one have been identified. These unsubstituted chromenones are equally potent at the 5-HT(1A) receptor and 5-HT transporter. The implementation of nitrogen functionalities in position 3 of the chromenones resulted in compounds acting as agonists at the 5 HT(1A) receptor and as 5-HT re-uptake inhibitors like vilazodone. Ex vivo 5-HT re uptake inhibition and in vitro 5-HT agonism were determined in the PCA- and GTPgammaS-assay, respectively. The potential of these chromenones to increase central 5-HT levels was measured in microdialysis studies and especially the derivatives 3-[4-[4-(3-amino-2-oxo-2H-chromen-6-yl)-piperazin-1-yl]-butyl]-1H indole-5-carbonitrile 8f, ethyl (6-[4-[4-(5-cyano-1H-indol-3-yl)-butyl]-piperazin 1-yl]-2-oxo-2H-chromen-3-yl)-carbamate 8h and N-(6-[4-[4-(5-cyano-1H-indol-3-yl) butyl]-piperazin-1-yl]-2-oxo-2H-chromen-3-yl)-acetamide 8k give rise to rapid development of increased serotonin levels in rat brain cortex, lasting longer than 3h. PMID- 15336264 TI - Photodynamic effects of porphyrin and chlorin photosensitizers in human colon adenocarcinoma cells. AB - Photodynamic therapy (PDT) is a cancer treatment involving systemic administration of a tumor-localizing photosensitizer; this, when activated by the appropriate wavelength of light, interacts with molecular oxygen to form a toxic, short-lived species known as singlet oxygen, which is thought to mediate cellular death. Photofrin, a complex mixture of porphyrin oligomers has recently received FDA approval for the photodynamic treatment of esophageal and endobronchial carcinoma, but its photodynamic and toxicity profiles are far from ideal. In the present study we evaluated a series of porphyrin-based PSs, some of which newly synthesized by our group, with the aim to identify agents with more favorable characteristics. For the most effective compounds in the porphyrin series, chlorin analogs were also synthesized; for comparison, the screening also included Photofrin. Cytotoxicity studies were performed by the MTT assay on a cultured human colon adenocarcinoma cell line (HCT116); the results indicate that the 3,4,5-trimethoxyphenyl, 3OH- and 4OH-phenyl, and the sulfonamidophenyl derivatives are significantly more potent than Photofrin. Flow cytometric studies and fluorescence microscopy indicate that in PDT-treated HCT116 cells death occurs mainly by apoptosis. In summary, novel PSs described in the present study, belonging both to the porphyrin and chlorin series, have proven more effective than Photofrin in killing colon cancer cells in vitro; extending these observation to in vivo models, particularly regarding the deeper reaching chlorin derivatives, might lead to significant advances in the development of tumor PDT. PMID- 15336265 TI - Synthesis of N-(beta-D-glucopyranosyl)- and N-(2-acetamido-2-deoxy-beta-D glucopyranosyl) amides as inhibitors of glycogen phosphorylase. AB - 2,3,4,6-Tetra-O-acetyl-beta-D-glucopyranosyl- and 2-acetamido-3,4,6-tri-O-acetyl 2-deoxy-beta-D-glucopyranosyl azides were transformed into the corresponding per O-acetylated N-(beta-D-glycopyranosyl) amides via a PMe(3) mediated Staudinger protocol (generation of N-(beta-D-glycopyranosyl)imino-trimethylphosphoranes followed by acylation with carboxylic acids, acid chlorides or anhydrides). The deprotected compounds obtained by Zemplen deacetylation were evaluated as inhibitors of rabbit muscle glycogen phosphorylase b. The best inhibitor of this series has been N-(beta-D-glucopyranosyl) 3-(2-naphthyl)-propenoic amide (K(i)=3.5microM). PMID- 15336266 TI - Anti-allergic activity of stilbenes from Korean rhubarb (Rheum undulatum L.): structure requirements for inhibition of antigen-induced degranulation and their effects on the release of TNF-alpha and IL-4 in RBL-2H3 cells. AB - Stilbenes isolated from the rhizomes of Rheum undulatum (Korean rhubarb) and the related compounds were investigated on their anti-allergic activities. The results revealed that 3,5,4'-trimethylpiceatannol exhibited the most potent inhibition against beta-hexosaminidase release as a marker of degranulation in RBL-2H3 cells with IC(50) of 2.1 microM, followed by trimethylresveratrol (IC(50)=5.1 microM). Structural requirements of stilbenes for the activity are as follows: (1) The oxygen functions (-OCH(3), -OH), especially methoxyl groups, are essential and their positions on aromatic rings are important for the activity; (2) the alpha-beta double bond increased the activity; (3) the glycoside moiety dramatically decreased the activity; and (4) the substitution group at the 3' position in trimethylresveratrol (3,5,4'-trimethoxystilbene) was preferably OH>H>OCH(3) for the activity. Several active stilbenes (piceatannol, 3,5,4' trimethylpiceatannol, resveratrol, trimethylresveratrol) also inhibited ionomycin induced beta-hexosaminidase release, suggesting that inhibition of Ca(2+) influx or degranulation mechanisms after Ca(2+) influx is important for their activities. Piceatannol, 3,5,4'-trimethylpiceatannol, resveratrol, and trimethylresveratrol also significantly inhibited antigen-induced release of TNF alpha and IL-4 in RBL-2H3 cells. PMID- 15336267 TI - New potent and selective A1 adenosine receptor agonists. AB - Thiirane analogs of ENAdo have been synthesised and found to be extremely potent and selective A(1) adenosine receptor agonists. PMID- 15336268 TI - In vitro activity and mechanism of action against the protozoan parasite Trypanosoma cruzi of 5-nitrofuryl containing thiosemicarbazones. AB - The in vitro growth inhibition activity of new thiosemicarbazone derivatives against the protozoan parasite Trypanosoma cruzi, the causative agent of American trypanosomiasis, are described. The designed compounds combine in the same molecule the thiosemicarbazone function, recently described as a potent cruzain inhibitor moiety, and the recognised 5-nitrofuryl group, an oxidative stress promoter. Some of the derivatives were found to be very active against the cultured (epimastigote) form of the parasite, being 1.5-1.7-fold more active than the reference compound, Nifurtimox. Free radicals production was detected when the compounds were incubated in presence of mammalian-liver microsomes. The thiosemicarbazones' capacity to act as pharmacophore in the cruzain inhibition process was theoretically analysed. Frontier molecular orbital HOMO was found as an adequate descriptor in this process. Acute in vivo toxicity of two of the more active derivatives was evaluated. The results showed that these compounds are among the most potent 5-nitrofuryl derivatives tested against this parasite thus support further in vivo studies of some of these thiosemicarbazones. PMID- 15336269 TI - Tricyclic oxazolo[2,3-f]purinediones: potency as adenosine receptor ligands and anticonvulsants. AB - Synthesis and physicochemical properties of 7-mono- and 6,7-disubstituted dihydrooxazolo-[3,2-f]purinediones are described. Oxazolo[2,3-f]purinediones were synthesized by cyclization of 8-bromotheophylline with oxiranes. The obtained compounds (1-22) were evaluated for their affinity at adenosine A(1) and A(2A) receptors. They showed mainly adenosine A(2A) receptor affinity at low micromolar concentrations and A(2A) selectivity, for example, compound 9 with an octyl substituent at the oxazole ring displayed adenosine A(2A) receptor affinity (K(i)=0.998 microM) and at least 25-fold A(2A) versus A(1) selectivity. This compound was less selective (5-fold) towards human recombinant A(2B) and A(3) adenosine receptors. In this group of compounds active adenosine A(1) receptor antagonists were also identified. Oxazolopurinediones were evaluated in vivo as anticonvulsants in MES and ScMet tests and examined for neurotoxicity in mice (ip). Compounds with long alkyl chains showed anticonvulsant activity in both tests (in 100 and 300 mg/kg doses), accompanied by significant neurotoxicity. The anticonvulsant activity in rats (po) was higher and without signs of neurotoxicity. SAR and QSAR studies stressed the importance of lipophilic 7 substituents for both types of pharmacological activity. The volume of the substituent is, however, limited at the A(2A) AR, an n-octyl group being optimal. PMID- 15336270 TI - Astersedifolioside A-C, three new oleane-type saponins with antiproliferative activity. AB - A phytochemical analysis of Aster sedifolius has led to the isolation of three novel triterpenoid saponins, based on an oleane-type skeleton and named astersedifolioside A (1), B (2) and C (3). On the basis of chemical, and 2D NMR and mass spectrometry data, the structures of the new compounds were elucidated as 3-O-[alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl] echinocystic acid 28-[O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranoside] (1), 3-O-[alpha L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl] echinocystic acid 28-[O-beta-D xylopyranosyl (1-->4)-O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L arabinopyranoside] (2) and 3-O-[alpha-L-rhamnopyranosyl (1-->2)-beta-D glucopyranosyl (1-->2)-beta-D-glucopyranosyl] echinocystic acid 28-[O-beta-D xylopyranosyl (1-->4)-O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L arabinopyranoside] (3). The isolated compounds showed antiproliferative effect in KiMol, a transformed thyroid cell line. PMID- 15336271 TI - Angiogenesis inhibitor TX-1898: syntheses of the enantiomers of sterically diverse haloacetylcarbamoyl-2-nitroimidazole hypoxic cell radiosensitizers. AB - (R)- and (S)-Epichlorohydrins were used to prepare the enantiomers of sterically diverse haloacetylcarbamoyl-2-nitroimidazoles that function as hypoxic cell radiosensitizers. The synthetic design allowed for introduction of a side chain of varying bulk that permitted an examination of the steric effects on enantio discrimination in biological assay systems. The single stereocenter also connected the two pharmacophores--a 2-nitroimidazole moiety critical to hypoxic cell radiosensitization, and a haloacetylcarbamoyl group to function as an anti angiogenesis pharmacophore. In the chick embryo chorioallantoic membrane (CAM) assay, the R-enantiomers possessing the bulky p-tert-butylphenyl group showed higher anti-angiogenic activity than the corresponding S-enantiomers, while there were no differences in the activity between the enantiomers containing the less bulky methyl and tert-butyl groups. Among the compounds we report, R-p-tert butylphenyl-bromoacetylcarbamoyl-2-nitroimidazole, TX-1898, was found to be the most promising candidate for further development of as anti-angiogenic hypoxic cell radiosensitizer. PMID- 15336272 TI - Synthesis and structure-activity relationships of the halovirs, antiviral natural products from a marine-derived fungus. AB - The halovirs are linear, lipophilic peptides produced by a marine-derived fungus of the genus Scytalidium. We recently reported that these molecules possess potent in vitro activity against the herpes simplex viruses 1 and 2. Here we present structure-activity relationships defining key structural elements for optimal viral inhibition. Results demonstrate that an N(alpha)-acyl chain of at least 14 carbons and an Aib-Pro dipeptide are critical for maintaining the antiviral activity. PMID- 15336273 TI - Structure-affinity studies for a novel series of homochiral naphtho and tetrahydronaphtho analogues of alpha 1 antagonist WB-4101. AB - A number of enantiomeric pairs of naphthodioxane, tetrahydronaphthodioxane and naphthoxy analogues of WB-4101 (1) were designed and synthesized in order to improve the selectivity profile of the parent compound, hopefully in favour of the alpha(1a)-AR with respect to the other two alpha(1) subtypes and the 5-HT(1A) receptor. The new compounds 2-8 and, in addition, the two enantiomers of 1 were tested in binding assays on the alpha(1a)-AR, alpha(1b)-AR, alpha(1d)-AR, and the 5-HT(1A) receptor. Two of them, namely the naphtho- and tetrahydronaphthodioxane derivatives (S)-2 and (S)-3, showed lower, but significantly more specific alpha(1a) affinity than (S)-1, while the two enantiomers of the 2-methoxy-1 naphthoxy analogue 6 maintained most of the very high alpha(1a) affinity of (S)-1 and its alpha(1a) versus alpha(1b) selectivity slightly increasing the alpha(1a)/alpha(1d) and alpha(1a)/5HT(1A) affinity ratios. The SAR data were evaluated in the light of known alpha(1) subtype pharmacophores and of the alpha(1a)-AR binding mode of WB-4101 resultant from literature mutagenesis studies disclosing some interesting consonances with these models. PMID- 15336274 TI - Synthesis and evaluation of phenylcarbamate derivatives as ligands for nicotinic acetylcholine receptors. AB - Phenylcarbamate derivatives were synthesized and evaluated in radioligand binding assays for different nicotinic acetylcholine receptor (nAChR) subtypes. Carbamate derivatives bearing a pyrrolidine or piperidine moiety 8-20 exhibited much lower affinity for alpha7* nAChR than the analogues in the quinuclidine series 21-25, although the same structural elements are present. Furthermore, in contrast to the quinuclidine analogues 21-25, all (S)-pyrrolidine derivatives 8-12 and the piperidine analogues 15 and 16 exhibited higher affinities for alpha4beta2* nAChR. PMID- 15336275 TI - Multidrug-resistant cancer cell susceptibility to cytotoxic quassinoids, and cancer chemopreventive effects of quassinoids and canthin alkaloids. AB - Twenty-three quassinoids (1-23), which were isolated previously from Simaroubaceous plants, were evaluated for cytotoxicity against three multidrug resistant cancer cell lines, KB-VIN, KB-7d, and KB-CPT. Nine compounds (2-7 and 9 11) showed significant cytotoxicity in all three cell lines. Compounds 1, 12-14, 17, and 20 demonstrated significant activity against the KB-7d and KB-CPT cell lines, and compounds 18, 19, and 23 revealed notable activity only against KB-7d cells. Structure-activity relationships were drawn based on these data. In addition, six quassinoid derivatives (24-29) and four canthin alkaloids (30-33), which were isolated from Brucea antidysenterica, were examined for their inhibitory effects on 12-O-tetradecanoylphorbol-13-acetate (TPA) induced Epstein Barr virus early antigen (EBV-EA) activation as cancer chemopreventive agents. All of these compounds demonstrated significant inhibitory effects against EBV-EA activation. PMID- 15336276 TI - Probing for a hydrophobic a binding register in prostate-specific membrane antigen with phenylalkylphosphonamidates. AB - To explore for the existence of an auxiliary hydrophobic binding register remote from the active site of PSMA a series of phenylalkylphosphonamidate derivatives of glutamic acid were synthesized and evaluated for their inhibitory potencies against PSMA. Both the phenyl- and benzylphosphonamidates (1a and 1b) exhibited only modest inhibitory potency against. The phenethyl analog 1c was intermediate in inhibitory potency while inhibitors possessing a longer alkyl tether from the phenyl ring, resulted in markedly improved K(i) values. The greatest inhibitory potency was obtained for the inhibitors in which the phenyl ring was extended furthest from the central phosphorus (1f, n=5 and 1g, n=6). The slightly serrated pattern that emerged as the alkyl tether increased from three to six methylene units suggests that inhibitory potency is not simply correlated to increased hydrophobicity imparted by the phenylalkyl chain, but rather that one or more hydrophobic binding registers may exist remote from the substrate recognition architecture in the active site of PSMA. PMID- 15336277 TI - Antibody-catalyzed oxidative degradation of nicotine using riboflavin. AB - Tobacco abuse remains a major cause of death worldwide despite ample evidence linking nicotine to various disease states. Consequently, immunopharmacotherapeutic approaches for the treatment of nicotine abuse have received increasing attention. Although a number of nicotine-binding antibodies have been disclosed, no antibody catalysts exist which efficiently degrade nicotine into pharmacologically inactive substances. Herein, we report the first catalytic antibodies which can oxidatively degrade nicotine. These biocatalysts use the micronutrient riboflavin and visible light as a source of singlet oxygen for the production of reactive oxygen species. Along with various known nicotine metabolites, antibody-catalyzed nicotine oxidations produce two novel nicotine oxidation products that were also detected in control ozonation reactions of nicotine. The reaction is efficient, with multiple turnovers of catalyst observed and total consumption of nicotine attained. These results demonstrate the potential of harnessing riboflavin as an endogenous sensitizer for antibody catalyzed oxidations and demonstrate a new approach for the development of an active vaccine for the treatment of nicotine addiction using in vivo catalytically active antibodies. PMID- 15336278 TI - The synthesis, distribution, and anti-hepatic cancer activity of YSL. AB - YSL was prepared stepwise from C terminal to N terminal with the side chain un protective amino acids, Boc-Leu-OMe, Boc-Ser-OH, and Boc-Tyr-OH, as the starting materials in 39.5% total yield (31.2g/per batch). With the side chain un protective Boc-(3,5-dibromo)-Tyr-OH and HCl.Ser-Leu-OMe as the starting materials (3,5-(3)H-Tyr)-Ser-Leu-OH was obtained in 29% yield. The determination of radioactive quantity in the urine and feces indicated that even after the administration for 130 h only 8.4% (5.35% in urine and 3.05% in feces) of total radioactive quantity from the metabolite of [3,5-(3)H-Tyr]-Ser-Leu-OH were monitored. The distribution study revealed the relative accumulation level of the individual tissue was arranged in the sequence of spleen>liver>kidney>lung>heart>muscle>brain. Selecting hepatic cancer as the target YSL significantly increased the survival time of H22 tumor cells implanted mice. PMID- 15336279 TI - Design, synthesis, and evaluation of aza inhibitors of chorismate mutase. AB - A series of aza inhibitors (4-9) of chorismate mutase (E.C. 5.4.99.5) was designed, prepared, and evaluated against the enzyme by monitoring the direct inhibition of the chorismate, 1, to prephenate, 2, conversion. None of these aza inhibitors displayed tighter binding to the enzyme than the native substrate chorismate or greater inhibitory action than the previously reported ether analogue, 3. Furthermore, no time-dependent loss of enzyme activity was observed in the presence of the two potentially reactive aza inhibitors (7 and 9). These results in conjunction with inhibition data from a broader series of chorismate mutase inhibitors allowed a novel proposal for the mechanistic role of chorismate mutase to be developed. This proposed mechanism was computationally verified and correlated with crystallographic studies of various chorismate mutases. PMID- 15336280 TI - Ambulatory system for the quantitative and qualitative analysis of gait and posture in chronic pain patients treated with spinal cord stimulation. AB - The physical activity in normal daily life is determined to a large extent by the functional ability of a subject. As a result, the measurement of the physical activity that a subject performs spontaneously could be a useful and objective measurement of disability, particularly in patients with disease-related functional impairment. The aim of this study is to provide an accurate method for the measurement and analysis of the physical activity under normal life conditions. Using three kinematical sensors strapped to the body, both the posture and the gait parameters can be assessed qualitatively and quantitatively. A detailed description of the algorithms used to analyse both the posture and the gait are presented in this paper. Two methods, based on different sensor configurations and signal processing, are proposed for the detection of sitting and standing postures (Methods P1 and P2). Two other methods are used for the quantitative assessment of walking (Methods W1 and W2). The performance of the algorithms (expressed in terms of sensitivity, specificity and error) is based on the comparison of data recorded simultaneously by a non-interfering observer (reference data) with the data provided by the recording system (21 patients, 61 h). Sensitivity and specificity are respectively 98.2% and 98.8% (P1), 97.8% and 98.1% (P2) for sitting; 98.0% and 98.5% (P1), 97.4% and 97.8% (P2) for standing; 97.1% and 97.9% (W1), 92.4% and 94.9% (W2) for walking; and finally, 99.2% and 98.6% for lying. Overall detection errors (as a percent of range) are as follows: 1.15% (P1) and 1.20% (P2) for sitting, 1.36% (P1) and 1.40% (P2) for standing, 1.20% (W1) and 1.60% (W2) for walking and 0.40% for lying. The error for the estimated walking distance and the speed is 6.8% and 9.6%, respectively. We conclude that both methods can be used for the accurate measurement of the basic physical activity in normal daily life. Measurements performed before and after the delivery of a treatment can therefore provide information of unprecedented accuracy and objectivity on the ability of a procedure, in this case spinal cord stimulation, to restore functional capabilities. PMID- 15336281 TI - Timing of muscle activity during reaching while standing: systematic changes with target distance. AB - We examined the effects of changing target distance from within arm's length (AL) to beyond arm's length on the onsets of electromyographic (EMG) activity of non focal muscles for a reaching task performed while standing. Two questions were addressed. First, do changes in target distance result in consistent changes in the onsets of non-focal anticipatory muscle activity of the trunk and legs in healthy subjects? Second, do changes in onsets of all non-focal muscles vary in a similar fashion in response to varying target distance? Thirteen young, healthy adults performed rapid, bilateral reaching movements to targets placed at shoulder height at four distances while electromyographic activity was recorded from muscles of the arm, trunk and legs. Ground reaction forces and arm kinematics were also recorded. The onsets of most non-focal muscles occurred prior to the onset of arm movement, and occurred progressively earlier as target distance was increased. An exception to this trend was the onset of the erector spinae muscle, which occurred progressively later as target distance was increased. These data support the notion that reaches to targets beyond arm's length involve anticipatory non-focal muscle activity that acts to transport the arm to the target rather than simply to resist the perturbation caused by the arm movement. The consistent patterns of anticipatory muscle activity observed in healthy subjects provide a template against which to compare activity patterns of non-focal muscles for individuals with potential deficits in the control of standing balance. PMID- 15336282 TI - The effect of velocity on the strategies used during gait termination. AB - The purpose of this study was to examine the changes that occur in gait termination (GT) as cadence increases. Five healthy volunteers walked at three cadences in time to a metronome under the following conditions: normal walking, planned stopping and unplanned stopping. Electromyography and force plate data were collected at 1000 Hz. Peak posterior ground reaction force (deceleration force) was greatest for the lead limb in unplanned stopping and, in general, deceleration force increased as cadence increased. Duration of muscle activity relative to the time to peak loading was unchanged by cadence and was always longest in the lead limb during planned stopping. As cadence increases, the signal to stop must occur earlier during stance phase if a person is to stop in the subsequent step. PMID- 15336283 TI - Magnitude and pattern of 3D kinematic and kinetic gait profiles in persons with stroke: relationship to walking speed. AB - The purpose of this study was to identify 3D kinematic and kinetic gait profiles in individuals with chronic stroke and to determine whether the magnitude or pattern (shape and direction of curve) of these profiles relate to gait performance (as measured by self-selected gait speed). More than one type of kinematic and kinetic pattern was identified in all three planes in 20 individuals with stroke (age: 61.2+/-8.4 years). Persons in the "fast" speed group did not necessarily exhibit the gait patterns closest to the ones reported for healthy adults. For example, in the frontal plane, a variation from the typical pattern (i.e., a hip abductor pattern in swing) was more common among the "fast" group. Correlations revealed that in addition to the sagittal profiles, the magnitudes of the frontal and transverse profiles are also related to speed, particularly the frontal hip powers. The results support the importance of hip abductors, in addition to the sagittal plane muscle groups, for both the paretic and non-paretic limbs. Furthermore, profiles which resemble gait patterns of neurologically healthy adults do not necessarily result in the faster gait speeds for individuals with chronic stroke. PMID- 15336284 TI - Relative motions of the tibia, talus, and calcaneus during the stance phase of gait: a cadaver study. AB - The motions of the tibia, talus, and calcaneus during walking were analyzed three dimensionally using a dynamic cadaver model that recreates the stance phase of walking. Rigid marker clusters were attached to each of the three bones, and the rotations of the talus and calcaneus with respect to the tibia and the calcaneus with respect to the talus were analyzed for eight right cadaver feet. The talus rotated primarily in plantarflexion/dorsiflexion about the talocrural joint, with an average range of 18 degrees +/- 4.7 degrees. The calcaneus began in inversion and internal rotation with respect to the tibia, moved into the neutral position at 28% of the stance phase and rotated primarily in plantarflexion from that point onward. Rotation of the calcaneus with respect to the talus at the subtalar joint occurred about all three axes, with approximately 5 degrees of relative dorsiflexion and 7 degrees of relative internal rotation. After 25% of stance, the talus and calcaneus moved together as one body into plantarflexion, providing a rigid lever as toe-off was approached. PMID- 15336285 TI - Diminished stepping responses lead to a fall following a novel slip induced during a sit-to-stand. AB - While neuromotor responses associated with successfully restoring balance have been widely characterized, little is known of how responses employed to regain balance fail. We identified biomechanical factors that characterized the unsuccessful recovery responses to a novel perturbation. An unexpected forward slip was induced, using a low-friction platform, just after seat-off of a sit-to stand in 41 young subjects. Kinematic variables describing the recovery responses were compared between fallers and those who recovered; "falls" into the fall arrest system were identified based on center of mass (COM) descent. Twelve "single-step" falls, four "multi-step" falls, and 11 recoveries resulted. Single step fallers exhibited a more posterior COM at slip onset, support (nonstepping) limb collapse resulting in hip vertical descent, and a shortened protective step backward. A factor analysis indicated that six common factors explained 88% of the variance in 22 kinematic variables describing the slip and recovery response. Single-step falls were associated with two factors, one related to initial or imposed instability and one to the reactive response. The former factor indicated that step length and COM position at step touchdown were related to COM position at slip onset. Support limb collapse comprised the latter factor. Multi-step fallers, by appearance, exhibited a shortened initial backward step that was ineffective at enabling balance recovery. As a whole, the results indicated that unsuccessful recovery was associated with a diminished stepping response. Fallers may have inadequately scaled an appropriate reflexive stepping response or might, inappropriately, have reflexively attempted to recover without stepping. PMID- 15336286 TI - Intra-abdominal pressure response to multidirectional support-surface translation. AB - A complex response of the trunk muscles occurs to restore equilibrium in response to movement of the support surface. Intra-abdominal pressure (IAP) is considered to contribute to control of the trunk. This study investigated the contribution of IAP to the postural response to multidirection support-surface translation. IAP was recorded with a thin-film pressure transducer inserted via the nose into the stomach and trunk motion was recorded with an optoelectronic system with markers over the spinous process of L1. A pattern of trunk movement was recorded in response to the support-surface translations that was consistent with a 'hip' strategy of postural control. The trunk moved in a manner appropriate to move the centre of gravity over the new base of support. IAP was increased with movement in each direction, but varied in timing and amplitude between translation directions. In general, the IAP was greater with translations in the sagittal plane compared to the frontal plane and was initiated earlier for translations in the backward direction. These data indicate that IAP contributes to the postural response associated with support-surface translation and suggest that this is consistent with stiffening the spine. PMID- 15336287 TI - Lower limb extensor moments in children with spastic diplegic cerebral palsy. AB - In this retrospective study, we quantified the mean extensor moment at the ankle, knee and hip over the stance period in a group of independently ambulant children with spastic diplegia (n = 90; 167 limbs) and in a group of normally-developing (ND) children (n = 22; 22 limbs). The mean knee extensor moment and the mean support moment demonstrated greater variance in children with diplegia than in normally-developing children (P < 0.0001 and P < 0.001). This was explained by a strong relationship between the mean knee extensor moment and minimum knee flexion in stance (r2 = 0.615; P < 0.0001) in the affected group with a positive mean knee extensor moment for all those children who walked in greater than 20 degrees of knee flexion. We also found a linear relationship between the support moment and knee flexion (r2 = 0.805; P < 0.0001). Our data supported the biomechanical analysis of Hof [Gait Posture, 12 (2000) 196] who suggested that his modified support moment should be a linear function with eccentricity at the knee. Extensor moments at the ankle (r2 = 0.001376; P = 0.641) and hip (r2 = 0.0860; P = 0.000168) bore weak relationships with increasing knee flexion even though there was a strong positive relationship between minimum knee flexion and minimum hip flexion (r2 = 0.316; P < 0.0001). We conclude that children with spastic diplegic cerebral palsy (SDCP) who walk with a crouch gait rely on their knee extensors to prevent collapse of the lower limbs. Intervention directed at redistributing extensor moments between the joints of the lower limbs may slow the increase in knee flexion and prolong reasonable walking function in this group. PMID- 15336288 TI - Three-dimensional realisation of muscle morphology and architecture using ultrasound. AB - Two-dimensional B-mode ultrasound imaging and motion tracking were combined to generate three-dimensional reconstructions of the medial gastrocnemius. Architectural and morphological features of this muscle could be visualised. The length of the gastrocnemius belly was measured in normally (ND) developing children and in children with spastic diplegic cerebral palsy (SDCP) who had plantarflexion contractures. Using a random effects linear model we demonstrated that the gastrocnemius muscle bellies of children with SDCP were shorter than those of ND children (P = 0.001) even when corrected for ankle position. The technique described could be used to evaluate muscular deformity before and after an intervention. PMID- 15336289 TI - A stair-climbing test for ambulatory assessment of children with cerebral palsy. AB - The purpose of this study was to develop a stair-climbing test to measure energy cost (EC) and mechanical efficiency (ME) in children with cerebral palsy (CP) to evaluate ambulation-related motor function and its changes after intervention or maturation. Five normally developed (ND) and 10 children with CP were tested. The gross ME (MEg) was calculated from the work done (W) and the total energy cost (oxygen consumption) measured while repeatedly ascending and descending four steps for approximately 5 min without subtracting the resting metabolic rate. The MEg was significantly lower in CP than ND (3% versus 20%, P < 0.001). The test was repeated in the 10 children with CP after a 4-month therapy recess. The MEg values correlated with the initial tests, with a small, significant increase of 2%. When calculating net ME (MEn) from W and the energy cost above resting, the correlation of MEn values before and after therapy was inferior to that using MEg values. Similarly, individual ME values obtained by estimating energy cost from the increase in heart rate (HR) during stair-climbing also correlated poorly, with large variability. These results show that MEg may be used to evaluate changes in motor function resulting from age-related development or therapy. MEg is as good or superior to MEn; the extra time required to obtain resting energy cost and heart rate values is not necessary when measurements are desired within the same individual. PMID- 15336290 TI - Postural reaction during maximum grasping maneuvers using a hand dynamometer in healthy subjects. AB - The purpose of this study was to quantitatively evaluate spontaneous body sway during maximum grasping maneuver with a hand dynamometer, which we sometimes encounter in clinical practice. The postural reaction of the whole body during maximum grasping maneuver was analyzed in 26 right-handed healthy male volunteers using a three-dimensional motion analysis system (VICON system, Oxford Metrics, UK) and force plates. We found that the gravity center of the body moved toward the grasping side. Also, voluntary rotation of the trunk and flexion of the neck was observed during grasping on each side. PMID- 15336291 TI - Measurement and management of errors in quantitative gait data. AB - Gait analysis is a valuable tool in the evaluation of children and adults with movement disorders. The data produced from gait analysis, however, is not necessarily free of errors. The purpose of this study was two-fold: (i) to estimate the errors associated with quantitative gait data; and (ii) to propose a method for incorporating the knowledge of these errors into the clinical interpretation process. An experimental protocol was designed that allowed within subject, within-observer and between-observer errors to be computed at each point in the gait cycle. The estimates were then used in a practical scheme for detecting significant deviations in joint angles. The results of this study provide a means for managing error, while simultaneously improving the rigor and objectivity of clinical interpretations. PMID- 15336292 TI - Statistical tools for clinical gait analysis. AB - Gait analysis studies involve continuous curves of data measured over a gait cycle. Curve analysis and interpretation require adequate statistical methods. Three principal problems may be encountered in clinical practice: (i) the reliability of gait curves for a given patient, (ii) classifying a new subject as belonging to a given population or not and (iii) comparison of two populations (independent or paired). This paper presents three statistical tools for solving these problems: (i) intra-class correlation coefficients, (ii) confidence bands for a population (iii) a combination of analysis of variance and confidence bands for the difference between the means and shows how they can be used in clinical practice. PMID- 15336293 TI - Repeatability of an optimised lower body model. AB - The optimisation technique, optimised lower-limb gait analysis (OLGA), is described together with a preliminary study of repeatability compared to an implementation of the Newington-Helen Hayes gait model. The study of repeatability used a single healthy subject, three physiotherapists as observers and provided approximately 100 gait cycles. Improvement in intra- and inter observer repeatability of the lower limb model was found for OLGA, indicated by significantly lower standard deviations (S.D.s) in local marker co-ordinate (a measure of rigidity of the marker attachment), together with reduced S.D. in the estimated length of the bone segments. The S.D. in the inter-hip distance measured by OLGA (N = 25) was found to be only 2.4 mm. The repeatability of clinically significant output variables (joint angles, forces and moments) was also improved, with the inter-observer variations for joint angles and forces being significantly lower for OLGA. Euler angle component cross-talk effects frequently reported at the hip, knee and ankle were also successfully reduced by OLGA, this being the chief cause of the improvement in inter-observer repeatability. PMID- 15336294 TI - A novel method for displaying gait and clinical movement analysis data. AB - Plotting kinematic and kinetic data of a patient's movement patterns relative to normative values (i.e., mean and +/-1 S.D.) is a common method used by clinicians to visually assess deviations and interpret the patient's gait analysis data. This method of data interpretation is often time consuming and complex, especially when the process requires the inspection of a plethora of line graphs for numerous variables that are displayed across several report pages. In this paper we propose an alternate method for displaying movement pattern deviations relative to normative data by color-coding the magnitude and the direction of the deviation. An advantage of this approach is that a single page summary of all the deviation magnitudes can be displayed simultaneously, in a manner that is concise, visually effective and reduces complexity. The purpose of this paper is to describe the algorithmic development of the color-coding method. PMID- 15336295 TI - Time-dependent changes in alcohol-seeking behaviour during abstinence. AB - Exposure of alcohol addicts to alcohol-related environmental cues may elicit alcohol-seeking behaviour even after protracted abstinence. The purpose of the present study was to assess time-dependent changes in alcohol-seeking behaviour in rats trained to respond for alcohol. The rats were re-exposed to alcohol associated stimuli after 1, 28 or 56 days of withdrawal. During the re-exposure session, the rats were first allowed to respond in extinction. Then, reinstatement of alcohol-seeking behaviour was evoked by a complex of discrete alcohol-associated cues (auditory and light cues combined with taste and smell of alcohol). Extinction behaviour depended on abstinence duration with maximal responding after 28-day abstinence. Reinstatement of alcohol-seeking behaviour evoked by the discrete cues was highest after 56-day abstinence. No correlations were found between individual alcohol intakes, extinction behaviour and cue induced reinstatement. These results suggest that: (i) alcohol-seeking behaviour may become more intense after long-term imposed abstinence; (ii) alcohol self administration, extinction behaviour, and reinstatement of alcohol-seeking behaviour may be regulated by separate neural mechanisms. PMID- 15336296 TI - Norharman and alcohol-dependency in male Wistar rats. AB - We examined the effects of ethanol ingestion to rats on levels of the beta carboline norharman in plasma, brain and liver at the end of ethanol ingestion and 10 h after withdrawal. We also investigated the effect of exogenously administered norharman on the behavioural signs of alcohol withdrawal. Ethanol was given by a liquid diet for 21 days. Norharman plasma levels in alcohol fed rats were significantly elevated compared to both control rats and to rats 10 h after withdrawal. Norharman levels in brains and livers showed a similar pattern. The capacity of the livers of both alcohol-dependent and withdrawal rats to catabolise norharman was significantly reduced compared to control rats. Norharman injected intraperitoneally (6.3 mg/kg) attenuated the behavioural signs of alcohol withdrawal significantly. The mechanism behind the increased norharman levels in alcohol-dependent rats may be inhibition of the synthesis and/or activity of liver enzyme(s) responsible for the breakdown of norharman. PMID- 15336297 TI - Using sleep to evaluate comparative serotonergic effects of paroxetine and citalopram. AB - BACKGROUND: SSRIs suppress rapid eye movement (REM) sleep, probably by increasing serotonin in the brainstem, and also increase sleep fragmentation. Although in the UK, paroxetine (PAR) and citalopram (CIT) have recommended doses of 20 mg/day for the treatment of depression, the recommended dose of CIT in USA is higher (40 mg). If similar doses of PAR and CIT have similar effects on central serotonin then they should have similar effects on sleep measures in volunteers. METHOD: This was a randomised, double blind placebo controlled crossover study in 12 healthy volunteers. Subjects took PAR 20 mg mane, CIT 20 mg mane or placebo mane for 3 days and sleep was recorded overnight at home on the third night. Standard measures of sleep were derived. RESULTS: REM sleep was significantly suppressed and sleep fragmentation increased by both drugs. Measures of REM sleep and sleep continuity previously found to be altered by SSRIs were considered together and compared with placebo as a 'serotonin response'; this was significantly greater in the PAR group. CONCLUSIONS: Sleep effects typical of SSRIs were greater with PAR 20 mg/day than CIT 20 mg/day, suggesting greater effects on 5HT uptake blockade. PMID- 15336298 TI - Prepulse inhibition in fawn-hooded rats: increased sensitivity to 5-HT1A receptor stimulation. AB - Prepulse inhibition of startle is a model of sensorimotor gating, which is disrupted in alcoholism, as well as mental illnesses such as schizophrenia. The fawn-hooded (FH) rat strain has been used as an animal model of alcoholism. FH rats showed significantly lower startle amplitude than Wistar-Kyoto (WKY) rats and Sprague-Dawley (SD) rats. Increasing doses of the 5-HT(1A) receptor agonist 8 OH-DPAT caused disruption of PPI, with the effect being significantly greater in FH rats compared to WKY rats. In all strains, treatment with 0.5 mg/kg of 8-OH DPAT significantly reduced PPI. In contrast, 0.1 mg/kg of 8-OH-DPAT caused disruption only in the FH strain. Treatment with amphetamine, apomorphine and MK 801 also significantly reduced PPI, however, there was no difference between the strains. This study shows increased sensitivity of FH rats to the disruption of PPI caused by 5-HT(1A) receptor stimulation, suggesting a link between 5-HT(1A) receptors, sensorimotor gating and aspects of the FH rat phenotype. PMID- 15336299 TI - Dieting and weight loss do not affect on the platelet serotonin 5-HT2A receptor. AB - Alterations related to the serotonin 5-HT(2A) receptor have been reported in various psychiatric disorders, and the 5-HT(2A) receptor is also one of the receptors mediating the effects of serotonin on feeding and satiety. The present study was carried out in order to investigate the association between the serotonin 5-HT(2A) receptor and weight loss during dieting in overweight subjects. In nine women studied before, during and after a 6-month period of dieting, body weight loss was not found to affect the platelet 5-HT(2A) receptor status. This finding implies that although body weight decrease is a common feature in many psychiatric disorders, the reported alterations in serotonin 5 HT(2A) receptor status in these disorders do not seem to be caused by the weight loss per se. PMID- 15336300 TI - Effects of the classical antipsychotic haloperidol and atypical antipsychotic risperidone on weight gain, the oestrous cycle and uterine weight in female rats. AB - Antipsychotic drug-induced side effects of weight gain and sexual dysfunction have clinical significance adversely affecting both compliance and morbidity. This study evaluated the effects of haloperidol and the atypical antipsychotic risperidone (0.1-1.0 mg/kg) on weight gain, food and water intake, the oestrous cycle and uterine weight in female hooded Lister rats. Haloperidol and risperidone treated rats displayed marked weight gain, although only risperidone induced significant increases in food consumption over the 21-day period. Neither haloperidol nor risperidone influenced water consumption. Marked disruption of the oestrous cycle was observed in risperidone- and haloperidol-treated animals (0.5 and 1.0 mg/kg), which was supported by significantly reduced uterine weights. The findings presented here suggest that the weight gain and sexual dysfunction induced by antipsychotics may be modelled in rodents. This model may offer insight into the mechanisms involved in mediation of such side effects. PMID- 15336301 TI - Stress and hippocampal abnormalities in psychiatric disorders. AB - The hippocampus plays a main role in regulating stress response in humans, but is itself highly sensitive to neurotoxic effects of repeated stressful episodes. Hippocampal atrophy related to experimental stress has been reported in laboratory studies in animals. Several controlled brain imaging studies have also shown hippocampal abnormalities in psychiatric disorders, including posttraumatic stress disorder (PTSD), major depressive disorder (MDD), and borderline personality disorder (BPD). This paper reviews the physiological role of the hippocampus in stress circuitry and the effects of stress on cognitive functions mediated by the hippocampus. We also review brain imaging studies investigating hippocampus in PTSD, MDD, and BPD. This literature suggests that individuals with PTSD, MDD, and BPD may suffer hippocampal atrophy as a result of stressors associated with these disorders. Prospective, longitudinal studies will be needed in high-risk offspring and first-episode subjects to explore the relationship between stress and hippocampal atrophy in these neuropsychiatric illnesses. PMID- 15336302 TI - Influence of the combined treatment of LY 300164 (an AMPA/kainate receptor antagonist) with adenosine receptor agonists on the electroconvulsive threshold in mice. AB - 7-Acetyl-5-(4-aminophenyl)-8,9-dihydro-8-methyl-7H-1,3-dioxolo-4,5H-2,3 benzodiazepine hydrochloride (LY 300164; a selective noncompetitive AMPA/kainate antagonist; 2 mg/kg) and N(6)-2-(4-aminophenyl)ethyl-adenosine (APNEA; a nonselective adenosine A(1)/A(3) receptor agonist; 2 and 3 mg/kg) significantly raised the threshold for electroconvulsions in mice. In contrast, 5'-N ethylcarboxamidoadenosine (NECA; a nonselective adenosine A(1)/A(2) receptor agonist; up to 1 mg/kg) did not affect the electroconvulsive threshold. The combined treatment of LY 300164 with NECA or APNEA was superior to single-drug medication as regards their protective action in this seizure model. Moreover, the combinations of LY 300164 with either NECA or APNEA were devoid of motor impairment, although they produced a significant long-term memory deficit. Measurement of the plasma levels of LY 300164 alone and in combination with APNEA or NECA did not suggest pharmacokinetic phenomena as an explanation for the interaction between these drugs. APNEA did not influence the plasma concentration of LY 300164. Moreover, NECA even significantly decreased the plasma levels of the AMPA/kainate antagonist. The present study clearly indicates a strong positive interaction in terms of anticonvulsant activity between LY 300164 and the drugs acting via adenosine receptors. PMID- 15336303 TI - Behavioural effects of rapid intravenous administration of meta chlorophenylpiperazine in patients with panic disorder and controls. AB - Oral and intravenous challenge paradigms with the direct 5-HT agonist meta chlorophenylpiperazine (m-CPP) in panic disorder (PD) have shown only moderate sensitivity or selectivity of the panicogenic effects in PD. However, the results of a study examining the effects of rapid intravenous administration of 0.1 mg/kg of m-CPP in healthy volunteers suggested that this approach may be a more selective and sensitive panicogenic paradigm in PD. We therefore compared the behavioural, neuroendocrine and physiological effects of rapid intravenous administration of 0.1 mg/kg of m-CPP in 10 patients with PD and 10 healthy controls. Panic attacks were significantly more provoked in patients with PD (90%) compared to healthy controls (0%). Effects on the behavioural, but not on the neuroendocrine and physiological parameters, were significantly greater in patients. Our data suggests that the behavioural effects of rapid intravenous administration of 0.1 mg/kg of m-CPP in patients with PD indeed show a unique combination of high sensitivity and selectivity. PMID- 15336304 TI - Intracerebroventricular injection of trazodone produces 5-HT receptor subtype mediated anti-nociception at the supraspinal and spinal levels. AB - Serotonin (5-HT) mediated anti-nociceptive effects induced by an anti-depressant, trazodone, are related to 5-HT(1A) receptor activities at the supraspinal level. 5-HT(3) receptor activation via the descending anti-nociceptive pathways may contribute to the trazodone mediated anti-nociception at the spinal level. Intracerebroventricular (i.c.v.) injection of trazodone dose-dependently impaired nociceptive responses in the formalin test in mice. Six and 15 microg of trazodone inhibited the early (P<0.05 or 0.01) and the late phases of the formalin test (P<0.05 or 0.01), while 3 microg had no effect. We examined the effects of a selective 5-HT(1A) receptor antagonist, WAY-100635, a single injection of which induced hyperalgesia (P<0.05), and blocked the anti nociceptive effects of trazodone (P<0.01) when the two were simultaneously injected i.c.v. Intrathecal (i.t.) injection of a selective 5-HT(3) receptor antagonist, 3-tropanylindole-3-carboxylate hydrochloride, blocked the anti nociceptive effects of i.c.v. trazodone (P<0.01), while WAY-100635 (i.t.) did not impair trazodone mediated anti-nociception. Trazodone mediated anti-nocicepton is related to serotonergic activity at both the supraspinal and the spinal level. PMID- 15336305 TI - ECNP Consensus Meeting, March 2003. Guidelines for the investigation of efficacy in social anxiety disorder. PMID- 15336307 TI - Signaling pathways controlling the production of inflammatory mediators in response to crystalline silica exposure: role of reactive oxygen/nitrogen species. AB - Occupational exposure to crystalline silica has been linked to pulmonary fibrosis and lung cancer. Surface properties of crystalline silica are critical to the production of oxidant species, chemokines, inflammatory cytokines, and proliferative factors involved in the initiation and progression of silica induced damage, inflammation, alveolar type II cell hyperplasia, fibroblast activation, and disease. The transcription factors nuclear factor kappaB (NF kappaB) and activator protein 1 (AP-1) have been shown to play key roles in gene promotion for inflammatory mediators, oncogenes, and growth factors. This review summarizes evidence that in vitro and in vivo exposure to crystalline silica results in activation of NF-kappaB and AP-1. Signaling pathways for activation of these transcription factors are described. In addition, the role of silica induced reactive oxygen species and nitric oxide in the activation of these signaling events is presented. Last, the generalizability of mechanisms regulating silica-induced pulmonary responses to pulmonary reactions to other occupational particles is discussed. PMID- 15336308 TI - Nitric oxide-releasing hydrophobic polymers: preparation, characterization, and potential biomedical applications. AB - The synthetic methods used recently in this laboratory to prepare a variety of novel nitric oxide (NO)-releasing hydrophobic polymers are reviewed. Nitric oxide is a well known inhibitor of platelet adhesion and activation. Thus, such NO release polymers have potential applications as thromboresistant coatings for a large number of blood-contacting biomedical devices (e.g., in vivo sensors, arteriovenous grafts, stents, catheters, extracorporeal circuits). The approaches taken to prepare NO releasing poly(vinyl chloride) (PVC), silicone rubber (SR), polymethacrylate (PM), and polyurethane (PU) materials are grouped into three categories: (1) dispersion/doping of discrete diazeniumdiolated molecules within the polymeric films; (2) chemical derivatization of polymeric filler microparticles (e.g., silicon dioxide, titanium dioxide) to possess NO release chemistry and then their dispersion within the hydrophobic polymers; and (3) covalent attachment of NO release moieties to polymer backbones. Specific chemical examples of each of these approaches are summarized and the advantages and disadvantages of each are discussed. Other related work in the field of NO release polymers is also cited. It is further shown that several of the NO releasing polymeric materials already prepared exhibit the expected improved thromboresistivity when tested in vivo using appropriate animal models. PMID- 15336309 TI - Reactions of 4-hydroxynonenal with proteins and cellular targets. AB - Peroxidative degradation of lipids yields the aldehyde 4-hydroxy-2-nonenal (4HNE) as a major product. The lipid aldehyde is an electrophile, and reactivity of 4HNE toward protein nucleophiles (i.e., Cys, His, and Lys) has been characterized. Through the use of purified enzymes and isolated cells, various pathways for biotransformation of the lipid aldehyde have been identified and include enzyme mediated oxidation, reduction, and glutathione conjugation. Uncontrolled oxidative stress can yield excessive lipid peroxidation and 4HNE generation, however, and overwhelm these cellular defenses. Indeed, in vitro and in vivo production of 4HNE in response to pro-oxidant exposure has been demonstrated using antibodies to protein adducts of the lipid aldehyde. Recent evidence suggests a role for protein modification by 4HNE in the pathogenesis of several diseases (e.g., alcohol-induced liver disease); however, the precise mechanism(s) is currently unknown but likely results from adduction of proteins involved in cellular homeostasis or biological signaling. PMID- 15336310 TI - The impact of carboxy nitroxide antioxidants on irradiated ataxia telangiectasia cells. AB - Three water-soluble carboxy nitroxide antioxidants, 5-carboxy-1,1,3,3 tetramethylisoindolin-2-yloxyl, 4-carboxy-2,2,6,6-tetramethylpiperidin-1-yloxyl, and 3-carboxy-2,2,5,5-tetramethylpyrrolidin-1-yloxyl, show significant impact on the postirradiation survival rates of ataxia telangiectasia (A-T) cells compared to normal cells, an assay which represents a model for understanding the impact of ROS damage on the A-T phenotype. The effects of these antioxidants are much more significant than those of vitamin E or Trolox (a water-soluble vitamin E analog), studied using the same cell survival model. PMID- 15336311 TI - Progressive iron accumulation induces a biphasic change in the glutathione content of neuroblastoma cells. AB - Glutathione (GSH) constitutes the single most important antioxidant in neurons, whereas iron causes oxidative stress that leads to cell damage and death. Although GSH and iron produce opposite effects on redox cell status, no mechanistic relationships between iron and GSH metabolism are known. In this work, we evaluated in SH-SY5Y neuroblastoma cells the effects of iron accumulation on intracellular GSH metabolism. After 2 d exposure to increasing concentrations of iron, cells underwent concentration-dependent iron accumulation and a biphasic change in intracellular GSH levels. Increasing iron from 1 to 5 microM resulted in a marked increase in intracellular oxidative stress and increased GSH levels. Increased GSH levels were due to increased synthesis. Further increases in iron concentration led to significant reduction in both reduced (GSH) and total (GSH + (2 x GSSG)) glutathione. Cell exposure to high iron concentrations (20-80 microM) was associated with a marked decrease in the GSH/GSSG molar ratio and the GSH half-cell reduction potential. Moreover, increasing iron from 40 to 80 microM resulted in loss of cell viability. Iron loading did not change GSH reductase activity but induced significant increases in GSH peroxidase and GSH transferase activities. The changes in GSH homeostasis reported here recapitulate several of those observed in Parkinson's disease substantia nigra. These results support a model by which progressive iron accumulation leads to a progressive decrease in GSH content and cell reduction potential, which finally results in impaired cell integrity. PMID- 15336312 TI - Noninvasive monitoring of glutathione turnover in perfused MCF-7 cells. AB - Glutathione metabolism was monitored in proliferating intact, perfused MCF-7 breast cancer cells by (13)C NMR spectroscopy. Label incorporation from [3,3' (13)C(2)]cystine in the perfusate into intracellular glutathione was monitored in native wild-type MCF-7 (MCF-7wt) cells and sublines resistant to doxorubicin (MCF 7dox) and 4-hydroperoxycyclophosphamide (MCF-7hc). Pulse-chase studies showed non linear rates of isotope label uptake and washout. Fitting these data to an exponential model of glutathione metabolism allowed calculation of rate constants for glutathione metabolism in these cell lines. Comparison of these rate constants showed glutathione metabolism was increased in both drug-resistant lines. No significant difference was observed between these results for cells growing in three dimensions and results for cells cultured in monolayer. PMID- 15336313 TI - Apomorphine prevents myocardial ischemia/reperfusion-induced oxidative stress in the rat heart. AB - This study examined the hypothesis that low-concentration apomorphine improves postischemic hemodynamic and mitochondrial function in the isolated rat heart model by attenuating oxidation of myocardial proteins. Control and apomorphine treated hearts were subjected to 35 min of perfusion, 25 min of normothermic global ischemia, and 60 min of reperfusion. Apomorphine (2 microM) was introduced into the perfusate for 20 min starting from the onset of reperfusion. Apomorphine significantly (p <.05) improved postischemic hemodynamic function: work index of the heart (product of LVDP and heart rate) was twice as high in apomorphine treated hearts compared to controls at the end of reperfusion (p <.01). After isolation of cardiac mitochondria, the respiratory control ratio (RCR) was calculated from the oxygen consumption rate of State 3 and State 4 respiration. Apomorphine significantly improved postischemic RCR (87% of preischemic value vs. 39% in control, p <.05). Using an immunoblot technique, carbonyl content of multiple unidentified myocardial proteins (mitochondrial and nonmitochondrial) was observed to be elevated after global ischemia and reperfusion. Apomorphine significantly attenuated the increased protein oxidation at the end of reperfusion. These results support the conclusion that apomorphine is capable of preventing ischemia/reperfusion-induced oxidative stress and thereby attenuating myocardial protein oxidation and preserving mitochondrial respiration function. PMID- 15336314 TI - Impaired superoxide radical production by bronchoalveolar lavage cells from NO(2) exposed rats. AB - Production of superoxide radicals is a central property of professional phagocytes used to combat invading microorganisms. Even though the number of macrophages and neutrophils is often increased in the lungs of patients with chronic lung diseases, these patients frequently suffer from bacterially induced exacerbations. To understand the underlying mechanisms, we investigated the production of superoxide radicals by bronchoalveolar lavage (BAL) cells in a rat NO(2) exposure model (10 ppm NO(2) for 1, 3, or 20 days). We showed that cells from NO(2)-exposed animals display a significantly impaired superoxide radical release after zymosan stimulation. The use of specific inhibitors (antimycin or diphenyleneiodonium [DPI]) revealed that the major enzyme systems, NADPH oxidase and complex III of the respiratory chain, are affected. In addition, we investigated gene expression and enzyme activities of antioxidant enzymes. mRNA expression was significantly enhanced for glutathione peroxidase (GPx)-3 and CuZn superoxide dismutase (SOD) in BAL cells from animals exposed 3 and 20 days, and GPx and SOD enzyme activities were increased in BAL cells from rats exposed 20 days. In conclusion, concomitant occurrence of reduced production and increased scavenging of superoxide radicals resulted in the drastically impaired release of these radicals from BAL cells of NO(2)-exposed rats. PMID- 15336315 TI - Superoxide dismutase and catalase are required to detect (.-)NO from both coupled and uncoupled neuronal no synthase. AB - Despite numerous approaches to measuring nitric oxide ((.-)NO) formation from purified NO synthase (NOS), it is still not clear whether (.-)NO is a direct or indirect product of the NO synthase reaction. The direct detection of catalytically formed (.-)NO is complicated by side reactions with reactive oxide species like H(2)O(2) and superoxide. The aim of the present study was therefore to reinvestigate these reactions both electrochemically and by chemiluminescence detection with particular emphasis on the requirement for cofactors and their interference with (.-)NO detection. Flavins were found to generate large amounts of H(2)O(2) and were therefore excluded from subsequent incubations. Under conditions of both coupled and uncoupled catalysis, SOD was absolutely required to detect (.-)NO from NOS. H(2)O(2) formation took place also in the presence of SOD and gave a smaller yet significant interfering signal. Similar data were obtained when the proposed intermediate N(omega)-hydroxy-l-arginine was utilized as substrate. In conclusion, standard Clark-type ()NO electrodes are cross sensitive to H(2)O(2) and therefore both SOD and catalase are absolutely required to specifically detect (.-)NO from NOS. PMID- 15336316 TI - Enhancement of cytotoxicity of artemisinins toward cancer cells by ferrous iron. AB - Iron(II) heme-mediated activation of the peroxide bond of artemisinins is thought to generate the radical oxygen species responsible for their antimalarial activity. We analyzed the role of ferrous iron in the cytotoxicity of artemisinins toward tumor cells. Iron(II)-glycine sulfate (Ferrosanol) and transferrin increased the cytotoxicity of free artesunate, artesunate microencapsulated in maltosyl-beta-cyclodextrin, and artemisinin toward CCRF-CEM leukemia and U373 astrocytoma cells 1.5- to 10.3-fold compared with that of artemisinins applied without iron. Growth inhibition by artesunate and ferrous iron correlated with induction of apoptosis. Cell cycle perturbations by artesunate and ferrous iron were not observed. Treatment of p53 wild-type TK6 and p53 mutated WTK1 lymphoblastic cells showed that mutational status of the tumor suppressor p53 did not influence sensitivity to artesunate. The effect of ferrous iron and transferrin was reversed by monoclonal antibody RVS10 against the transferrin receptor (TfR), which competes with transferrin for binding to TfR. CCRF-CEM and U373 cells expressed TfR in 95 and 48% of the cell population, respectively, whereas TfR expression in peripheral mononuclear blood cells of four healthy donors was confined to 0.4-1.3%. This indicates that artemisinins plus ferrous iron may affect tumor cells more than normal cells. The IC(50) values for a series of eight different artemisinin derivatives in 60 cell lines of the U.S. National Cancer Institute were correlated with the microarray mRNA expression of 12 genes involved in iron uptake and metabolism by Kendall's tau test to identify iron-responsive cellular factors enhancing the activity of artemisinins. This pointed to mitochondrial aconitase and ceruloplasmin (ferroxidase). PMID- 15336317 TI - Nitric oxide and pulmonary arterial pressures in pulmonary hypertension. AB - Decreased production of vasodilator substances such as nitric oxide (NO) has been proposed as important in development of pulmonary arterial hypertension (PAH). We hypothesize that NO measured over time serves as a non invasive marker of severity of PAH and response to therapy. We prospectively and serially measured exhaled NO and carbon monoxide (CO), a vasodilator and anti-inflammatory product of heme oxygenases, in 17 PAH patients in conjunction with hemodynamic parameters over 2 years. Although pulmonary artery pressures and NO were similar in all patients at entry to the study, NO increased in the 12 individuals who survived to complete the study, and correlated with change in pulmonary artery pressures. In contrast, CO did not change or correlate with hemodynamic parameters. Investigation of NO-oxidant reaction products in PAH in comparison to controls suggests that NO synthesis is impaired in the lung and that reactive oxygen species may be involved in the pathophysiology of pulmonary hypertension. Endogenous NO is inversely related to pulmonary artery pressure in PAH, with successful therapy of PAH associated with increase in NO. PMID- 15336318 TI - Is serum gamma-glutamyltransferase inversely associated with serum antioxidants as a marker of oxidative stress? AB - A series of studies in black and white women and men have suggested that serum gamma-glutamyltransferase (GGT) within its normal range might be an early marker of oxidative stress. If serum GGT is a marker of oxidative stress, it might have important implications both clinically and epidemiologically because measurement of serum GGT is easy, reliable, and not expensive. We examined the cross sectional association between deciles of serum GGT and concentrations of serum antioxidants among 9083 adult participants in the third U.S. National Health and Nutrition Examination Survey. After adjustment for race, sex, age, and total cholesterol, serum concentration of GGT across all deciles was inversely associated with serum concentrations of alpha-carotene, beta-carotene, beta cryptoxanthin, zeaxanthin/lutein, lycopene, and vitamin C (p for trend <.01, respectively). Vitamin E was not associated with serum GGT. All these associations were not materially different after additional adjustment for total energy intake, body mass index, smoking status, smoking amount, alcohol intake, and exercise. These associations were similarly observed among most subgroups. In conclusion, the current and previous studies strongly suggest that serum GGT level within its normal range may be an early marker of oxidative stress. PMID- 15336319 TI - Nuclear factor kappaB and activating protein 1 are involved in differentiation related resistance to oxidative stress in skeletal muscle cells. AB - Skeletal muscle cells are continuously exposed to oxidative stress. Thus, they compensate environmental challenges by increasing adaptive responses, characterized by activating protein 1 (AP-1)- and nuclear factor kappaB (NF kappaB)-mediated transcriptional upregulation of endogenous enzymatic and nonenzymatic antioxidants. We investigated the crosstalk of molecules involved in redox signaling in muscle cells, by using the rat L6C5 and mouse C2C12 cell lines, which represent a useful experimental model for studying muscle metabolism. We analyzed specific antioxidant systems, including glutathione, thioredoxin reductase, and antioxidant enzymes, and the redox-sensitive transcription factors AP-1 and NF-kappaB, in both myoblasts and myotubes. We found that the high levels of NF-kappaB DNA binding activity and thioredoxin reductase, together with inhibitory AP-1 complexes, allowed increased expression of antioxidant enzymes and survival of C2C12 cells after oxidant exposure. On the contrary, L6C5 myoblasts had a sensitive phenotype, correlated with lower levels of thioredoxin reductase, catalase, and NF-kappaB activity and higher levels of GSSG and activating AP-1 complexes. Interestingly, this cell line acquired an apoptosis-resistant phenotype, accompanied by drastic changes in the oxidant/antioxidant balance, when induced to differentiate. In conclusion, the two cell lines, although similar in terms of growth and differentiation, displayed significant heterogeneity in terms of redox homeostasis. PMID- 15336320 TI - The effects of meso-2,3-dimercaptosuccinic acid and oligomeric procyanidins on acute lead neurotoxicity in rat hippocampus. AB - Oxidative stress is considered to be a mechanism involved in lead neurotoxicity. Apoptosis is also thought to relate to lead neurotoxicity. The present study, focused on the hippocampus, was designed to investigate the two possible mechanisms involved in lead neurotoxicity and the potential protective effects of 2,3-dimercaptosuccinic acid (DMSA) and oligomeric procyanidins (OPC). It was proved that reactive oxygen species and oxidative damage were implicated in the induction of apoptosis induced by lead in the hippocampus. Administration of DMSA attenuated the oxidative stress and apoptosis in addition to having strong chelating and lead-removing capacity. OPC alone had antioxidant protective effects in the hippocampus but no removing capacity for lead in vivo despite showing higher affinity and stronger chelating ability for Pb(2+) than DMSA in vitro. It is suggested that OPC chelates Pb(2+) but does not discharge it from the body and even accumulates Pb(2+) in some organs. At the same time, a reasonable deduction can also be made that the complex of OPC-Pb(2+) prevents or at least weakens the neurotoxicity of Pb(2+). Whether this complex displays toxicity over a long time span should be studied further. PMID- 15336321 TI - Redox state alteration modulates astrocyte glucuronidation. AB - We have investigated the effects of mild oxidative conditions on drug metabolizing enzyme activity in rat cultured astrocytes. These experimental conditions promoting an oxidative environment were obtained by short exposure to a low concentration of menadione (5 microM) for a short duration (15 min). This resulted in the rapid and transient production of reactive oxygen species (+130%), associated with a decrease in GSH cellular content (-24%), and an increase in total protein oxidation (+26%), but promoted neither PGE(2) nor NO production. This treatment induced a rapid and persistent decrease in astrocyte glucuronidation activities, which was totally prevented by N-acetyl-l-cysteine. These oxidative conditions also affected the specific UGT1A6 activity measured in transfected V79-1A6 cells. Finally, the subsequent recovery of astrocyte glucuronidation activity may result from upregulation of UGT1A6 expression (+62%) as shown by RT-PCR and gene reporter assay. These results show that the catalytic properties and expression of cerebral UGT1A6 are highly sensitive to the redox environment. The protective effect of N-acetyl-l-cysteine suggests both a direct action of reactive oxygen species on the protein and a more delayed action on the transcriptional regulation of UGT1A6. These results suggest that cerebral metabolism can be altered by physiological or pathological redox modifications. PMID- 15336322 TI - Release of reactive oxygen and nitrogen species from contracting skeletal muscle cells. AB - A number of studies have indicated that exercise is associated with an increased oxidative stress in skeletal muscle tissue, but the nature of the increased oxidants and sites of their generation have not been clarified. The generation of extracellular reactive oxygen and nitrogen species has been studied in myotubes derived from an immortalized muscle cell line (H-2k(b) cells) that were stimulated to contract by electrical stimulation in culture. Cells were stimulated to contract with differing frequencies of electrical stimulation. Both induced release of superoxide anion and nitric oxide into the extracellular medium and caused an increase in extracellular hydroxyl radical activity. Increasing frequency of stimulation increased the nitric oxide generation and hydroxyl radical activity, but had no significant effect on the superoxide released. Additions of inhibitors of putative generating pathways indicated that contraction-induced NO release was primarily from neuronal NO synthase enzymes and that the superoxide released is likely to be generated by a plasma membrane located, flavoprotein oxidoreductase system. The data also indicate that peroxynitrite is generated in the extracellular fluid of muscle during contractile activity. PMID- 15336323 TI - Menadione induces a low conductance state of the mitochondrial inner membrane sensitive to bongkrekic acid. AB - When rat liver mitochondria are allowed to cycle Ca(2+) and are incubated in the presence of the pro-oxidant menadione, they undergo swelling, membrane potential (DeltaPsi) collapse, and ion release. These effects, which are inhibited by cyclosporin A (CsA), are fully consistent with the opening of the so-called permeability transition pore. However, when Ca(2+) cycling is abolished by EGTA, the mitochondria remain energized (DeltaPsi collapse and swelling are avoided), but Ca(2+) efflux, promoted by the chelating agent, is stimulated by menadione. This stimulation goes together with the release of Mg(2+), K(+), and adenine nucleotides (AdN) and is inhibited by bongkrekic acid (BKA). The effect of menadione is also characterized by biphasic NAD(P)H oxidation which becomes monophasic in the presence of BKA, CsA, or EGTA and by the oxidation of thiol groups not restrained by the above-mentioned inhibitors. These results suggest that BKA acts indirectly by preserving in the matrix a critical amount of AdN without modifying the monophasic oxidation of pyridine nucleotides by menadione. A critical number of thiol groups also seems to be involved in the phenomenon. Their oxidation most probably causes a conformational change on adenine nucleotide translocase with the opening of the "low-conductance state" of the mitochondrial permeability transition, resulting in ion permeability without DeltaPsi disruption and mitochondrial swelling. PMID- 15336324 TI - Reactive oxygen species in choline deficiency-induced apoptosis in rat hepatocytes. AB - Choline deficiency (CD) is involved in hepatocellular carcinoma and CD-induced apoptosis may be implicated in cellular malignant transformation. In this report, we studied the effects of choline deficiency on generation of reactive oxygen species (ROS) using the fluorescent probe dichlorodihydrofluorescein diacetate and the possible role of ROS on CD-induced apoptosis in cultured CWSV-1 cells, an immortalized rat hepatocyte. This cell line is reported to become tumorigenic by step-wise culturing in lower levels of choline. Our data demonstrate that CD induces a time- and dose-dependent increase in ROS in CWSV-1 cells. The increase in ROS production may be related to dysfunction of the mitochondrial respiratory chain. Our data also demonstrated that ROS generation occurred before CD-induced apoptosis, suggesting ROS may play a key role in signaling CD-induced apoptosis in CWSV-1 cells. PMID- 15336326 TI - Re: systematic review of long-acting oral opioids. PMID- 15336328 TI - Target therapies in palliative care: from a clinical to a biological approach. PMID- 15336330 TI - Re: anemia at the end of life. PMID- 15336332 TI - Medicare cost in matched hospice and non-hospice cohorts. AB - Hospice care is perceived as enhancing life quality for patients with advanced, incurable illness, but cost comparisons to non-hospice patients are difficult to make. The very large Medicare expenditures for care given during the end of life, combined with the pressure on Medicare spending, make this information important. We sought to identify cost differences between patients who do and do not elect to receive Medicare-paid hospice benefits. We introduce an innovative prospective/retrospective case-control method that we used to study 8,700 patients from a sample of 5% of the entire Medicare beneficiary population for 1999-2000 associated with 16 narrowly defined indicative markers. For the majority of cohorts, mean and median Medicare costs were lower for patients enrolled in hospice care. The lower costs were not associated with shorter duration until death. For important terminal medical conditions, including non cancers, costs are lower for patients receiving hospice care. The lower cost is not associated with shorter time until death, and appears to be associated with longer mean time until death. PMID- 15336333 TI - Causal attributions for fatigue among late middle-aged and older adults with HIV infection. AB - Fatigue, a symptom with many possible causes, is highly prevalent among persons with HIV/AIDS. The causal attributions HIV+ individuals make for their fatigue are important because they largely guide the coping responses they use to manage it. In a study to better understand the symptom appraisal process among 100 HIV+ older adults (50+), 49 participants discussed in depth their experience with fatigue. Many participants felt confused, at least initially, about the source of their fatigue, although most offered their HIV disease as a possible cause. However, many who did so also believed it was caused in part by other, generally less threatening causes, such as age, overexertion, or excess weight. HIV medications and other symptoms or comorbidities were also frequently cited as causes of fatigue. Health care professionals and HIV+ peers often helped to shape or reinforce participants' attributions for their fatigue. PMID- 15336334 TI - No evidence for sex differences in the severity and treatment of cancer pain. AB - While chronic pain is experienced by approximately 50-90% of patients with metastatic cancer, little is known about sex differences in chronic cancer pain. Therefore, the purposes of this study, in a sample of oncology outpatients (n=187) who were experiencing pain from bone metastasis, were: 1) to determine if there were sex differences in various pain characteristics, including pain intensity, and 2) to determine if there were sex differences in the prescription and consumption of analgesic medications. No significant sex differences were found in any of the baseline pain characteristics. In addition, no significant sex differences were found in analgesic prescriptions or intake of analgesic medications. Of note, men reported significantly higher pain interference scores for sexual activity than women. The study findings are important because they suggest that, unlike in acute pain, sex may not influence patients' perceptions of and responses to chronic cancer pain. PMID- 15336335 TI - Palliative care units in The Netherlands: changes in patients' functional status and symptoms. AB - Although efforts have been made to define optimal terminal care in palliative care units (PCUs), comprehensive longitudinal evaluations of care outcomes in PCUs at the end of life are scarce. In this study, changes in functional status (assistance needed for walking, and toilet use) and symptoms (pain, nausea, shortness of breath, depression, and anxiety) were assessed in all patients (n=355) admitted to 10 PCUs in Dutch nursing homes. Outcomes were measured at 24 hours, 48 hours, one week, and two weeks before death, and at PCU admission. Results show that functional status deteriorated from admission to one week before death, but most symptoms did not worsen in the last three weeks before death. Decreases in pain, anxiety, and nausea were observed. The results suggest that the care provided in the PCUs stabilized the symptom levels. Patients who die between two and four weeks appeared to have more favorable symptom change patterns than patients who die within two weeks, which supports the recommendation to admit eligible patients in earlier phases of their disease. Limitations include the use of proxy measures and some forms of selection bias, which may lead to underestimation of symptom levels. PMID- 15336336 TI - Massage therapy for symptom control: outcome study at a major cancer center. AB - Massage is increasingly applied to relieve symptoms in patients with cancer. This practice is supported by evidence from small randomized trials. No study has examined massage therapy outcome in a large group of patients. At Memorial Sloan Kettering Cancer Center, patients report symptom severity pre- and post-massage therapy using 0-10 rating scales of pain, fatigue, stress/anxiety, nausea, depression and "other." Changes in symptom scores and the modifying effects of patient status (in- or outpatient) and type of massage were analyzed. Over a three-year period, 1,290 patients were treated. Symptom scores were reduced by approximately 50%, even for patients reporting high baseline scores. Outpatients improved about 10% more than inpatients. Benefits persisted, with outpatients experiencing no return toward baseline scores throughout the duration of 48-hour follow-up. These data indicate that massage therapy is associated with substantive improvement in cancer patients' symptom scores. PMID- 15336337 TI - Predicting aberrant drug behavior in patients treated for chronic pain: importance of abuse history. AB - Physicians can encounter problems in prescribing opioids for some patients with chronic pain such as multiple unsanctioned dose escalations, episodes of lost or stolen prescriptions, and positive urine drug screenings for illicit substances. This study explored the usefulness of questions on abuse history in predicting problems with prescribing opioids for patients at a hospital-based pain management program. One hundred forty-five (145) patients who were taking long- and short-acting opioids for their pain were classified as high or low risk on the basis of their responses to interview questions about 1) substance abuse history in their family, 2) past problems with drug or alcohol abuse, and 3) history of legal problems. The treating physicians completed a questionnaire about problems that they had encountered with their patients. Problem behaviors were verified through chart review. No differences in demographic characteristics were found between those classified as high and low risk. Patients who admitted to a family history of substance abuse, a history of legal problems, and drug or alcohol abuse were prone to more aberrant drug-related behaviors, including a higher incidence of lost or stolen prescriptions and the presence of illicit substances in their urine (P < 0.05). Patients classified as high risk also had a significantly higher frequency of reported mental health problems and motor vehicle accidents. More of these patients smoked cigarettes, tended to need a cigarette within the first hour of the day, took higher doses of opioids, and reported fewer adverse effects from the medications than did those without such a history (P < 0.05). This study demonstrates that questions about abuse history and legal problems can be useful in predicting aberrant drug-related behavior with opioid use in persons with chronic noncancer pain. PMID- 15336338 TI - Electronic pain diary: a randomized crossover study. AB - Electronic pain diaries and palm-top computers have become increasingly important in clinical research and practice. In a randomized crossover trial, 24 patients suffering from chronic cancer and non-cancer pain completed both the electronic and the paper version of a pain diary based on the Minimal Documentation System (MIDOS) for pain and symptom assessment. This includes daily assessment of pain on an 11-point numeric rating scale and weekly documentation of a short quality of-life questionnaire. Of 52 patients seen during the baseline phase of this study, 28 could be enrolled and only 24 patients completed both diary versions. The other patients were either physically or intellectually unable to use a palm top computer or unwilling to participate in this study. After a total of four weeks, patient satisfaction was remarkably higher for the electronic palm-top version, even though a high number of patients were lacking experience in the use of computers. Obvious differences were observed between the versions. There were higher numbers of missing values in the electronic data, and patients tended to retrospectively fabricate information in the paper version. No significant difference between the electronic and paper diary could be found assessing the documented pain and symptom intensity. The electronic diary was used more frequently and patients said its use supported a more regular pharmacotherapy. We conclude that the use of electronic pain diaries is a valid and feasible method for documenting patients' pain perception, though some patients may not be able to operate such a diary version. Electronic palm-top pain diaries provide a high degree of patient satisfaction and can ease data collection for clinical research and practice. PMID- 15336339 TI - Drug stability testing to support clinical feasibility investigations for intrathecal baclofen-clonidine admixture. PMID- 15336340 TI - Massive hydromorphone dose delivered subcutaneously instead of intrathecally: guidelines for prevention and management of opioid, local anesthetic, and clonidine overdose. AB - Increasing numbers of patients with refractory pain are receiving intrathecal drug delivery systems (IDDS). We describe a case to illustrate the clinical manifestations and management implications of inadvertent overdose with drugs used in IDDS, including opioids, clonidine, baclofen, and local anesthetics. An IDDS patient received a bimonthly dose of intrathecal hydromorphone subcutaneously. The patient received a total of 540 mg hydromorphone into the subcutaneous pocket around the intrathecal pump. She was treated with naloxone intravenously over 12 hours, and had no major adverse sequelae. Such occurrences may happen more frequently with the expanded use of IDDS and clinicians should be prepared to take quick action. Counteracting an opioid with naloxone until the opioid is metabolized and excreted can be done safely. Inadvertent subcutaneous administration of other types of drugs could be more difficult to manage. PMID- 15336341 TI - The effect of transdermal fentanyl treatment on serum cortisol concentrations in patients with non-cancer pain. AB - We treated 50 patients with chronic nonmalignant pain using transdermal fentanyl (TDF) 25 microg/hr and concurrently measured pain using a visual analog scale (VAS) and serum cortisol concentration. We determined these outcomes at baseline and on days 30, 60, and 90 of the therapy. The patients also were asked to document any adverse effects. We found that mean cortisol concentrations on days 30, 60, and 90 of therapy were significantly (P < 0.0001) lower than the basal mean cortisol level, and mean VAS scores at days 30, 60, and 90 of therapy were also significantly better than the initial mean value (P < 0.0001). Fourteen patients experienced severe adverse events. These observations suggest that serum cortisol concentrations may be elevated in chronic non-cancer pain states and that TDF therapy can reduce cortisol levels in parallel with reduction in pain. PMID- 15336342 TI - The management of malignant psoas syndrome: case reports and literature review. AB - Malignant psoas syndrome (MPS) was first described in 1990, and is characterized by proximal lumbosacral plexopathy, painful fixed flexion of the ipsilateral hip, and radiological or pathological evidence of ipsilateral psoas major muscle malignant involvement. There have been 23 case reports of MPS in medical journals. Despite being associated with a severe and difficult pain, there is no definitive approach to management presented in the palliative care literature. We review the relevant clinical features and the subsequent multidisciplinary pain management in relation to four new cases of malignant involvement of the psoas muscle, and the 23 case reports in the literature. We propose that MPS comprises a continuum of symptoms and signs related to the degree of anatomical destruction with associated inflammatory reaction and muscle spasm, and also the degree of lumbosacral plexopathy causing neuropathic pain. A protocol is presented for the management of the complex pain issues of MPS directed at likely mechanisms. The treatment options include opioids, agents for neuropathic pain, muscle relaxants to manage psoas muscle spasm, and anti-inflammatory agents to reduce peritumoral edema. Direct anti-tumor measures also need to be considered. Further prospective study is needed to validate the proposed methods of assessment and treatment. PMID- 15336343 TI - Improving referral of patients to hospice through community physician outreach. PMID- 15336344 TI - ESR investigation of gamma irradiated sulbactam sodium. AB - In the present work, radiolysis of gamma irradiated sulbactam sodium (SS) was investigated through detailed ESR studies performed at low and at high temperatures on the radiolytic products induced in this drug after gamma irradiation in the dose range of 3-15 kGy. While unirradiated SS presented no ESR signal, irradiated SS exhibited an ESR spectrum with many resonance peaks. Variations of the spectrum pattern and of the intensities of resonance peaks with microwave power, applied radiation dose, storage time and temperature were followed. A radical with unpaired electron localised on the carbon atom of five membered ring directly attached to two CH(3) groups and SO(2)(-) ionic radical called as species A and B, respectively, were found to be produced after gamma radiolysis of SS. The characteristic spectral and kinetic features of these species were determined by spectrum simulation calculation and by curve fitting techniques using experimental signal intensity data as input. Although, species A was fairly unstable at room and especially at high temperatures, species B were observed to be relatively stable even at high temperatures having activation energies of E(A) (+/-4)kJ/mol and E(B) (+/-3)kJ/mol, respectively, at room temperature. Four different mathematical functions were tried to explore the dosimetric features of SS and a linear function of applied dose described best the dose-response data obtained for stable species, that is, radical B, in the dose range of 0-10 kGy. An accuracy of 4% in the dose measurements was concluded to be achievable through back-projection dose calculations if SS were used as a dosimetric material. PMID- 15336345 TI - Development of a robust procedure for assessing powder flow using a commercial avalanche testing instrument. AB - The objectives of this work were to develop a robust procedure for assessing powder flow using a commercial avalanche testing instrument and to define the limits of its performance. To achieve this a series of powdered pharmaceutical excipients with a wide range of flow properties was characterized using such an instrument (Aeroflow, TSI Inc., St. Paul, MN, USA). The experimental conditions (e.g., sample size, rotation speed) were rationally selected and systematically evaluated so that an optimal standard-operating-procedure could be identified. To evaluate the inherent variability of the proposed methodology samples were tested at multiple sites, using different instruments and operators. The ranking of the flow properties of the powders obtained was also compared with that obtained using a conventional shear-cell test. As a result of these experiments a quick, simple, and rugged procedure for determining the flow properties of pharmaceutical powders in their dilated state was developed. This procedure gave comparable results when performed at four different testing sites and was able to reproducibly rank the flow properties of a series of common pharmaceutical excipient powders. The limits of the test method to discriminate between different powder samples were determined, and a positive correlation with the results of a benchmark method (the simplified shear cell) was obtained. PMID- 15336346 TI - Measurement of triclabendazole and its metabolites in liver flukes: method development and full validation. AB - The trematode parasite Fasciola hepatica is still the cause of considerable loss in livestock production all over the world. The knowledge of the pharmacological properties of the available flukicidal drugs is critical to control this parasite. Triclabendazole is an halogenated benzimidazole anthelmintic with potent flukicidal activity. A simple reversed-phase high-performance liquid chromatographic analytical method has been developed, validated and applied for the quantitative determination of the flukicidal drug triclabendazole and its sulpho- and hydroxy-metabolites in the liver fluke, F. hepatica. Triclabendazole, triclabendazole sulphoxide, triclabendazole sulphone, hydroxy-triclabendazole, hydroxy-triclabendazole sulphoxide and hydroxy-triclabendazole sulphone were measured simultaneously in this trematode parasite. Linearity, resolution, precision, accuracy, recovery, limits of detection and quantification of the method were determined. Drug extraction from the parasite's tissue homogenate was effectively performed using liquid extraction (acetonitrile), avoiding clean up by solid phase extraction, prior to analysis by reversed-phase high-performance liquid chromatography. The resolution of all the halogenated benzimidazole thiol molecules assayed was obtained on a C(18) reversed-phase (5 microm, 250 mm x 4.6 mm) column using acetonitrile and ammonium acetate as the mobile phase and ultraviolet detection at 300 nm. Regression analyses were linear over the concentration range examined (from 0.272 to 16.331 nmol/100 mg trematode protein) and the correlation coefficients of the calibration curves ranged between 0.996 and 1.000. The calculated limits of detection of the proposed method for the parent drug and its metabolites ranged between 0.007 and 0.079 nmol/100 mg trematode protein. The extraction efficiency for the different analytes from the parasite material was greater than 71%. The results obtained indicated that the developed chromatographic method was selective, accurate and easy to reproduce. The developed procedure was successfully applied to quantify triclabendazole/metabolites in F. hepatica incubated under ex vivo conditions, demonstrating to be efficient for the determination of the most extensively used flukicidal drug available for veterinary medicine, and its metabolites. The analytical method described here is an useful tool for the measurement of this flukicidal compound in different studies addresses to evaluate drug influx/eflux and metabolism in its main target parasite. PMID- 15336347 TI - Electrochemical determination of Cephalothin antibiotic by adsorptive stripping voltammetric technique. AB - A sensitive and reliable stripping voltammetric method was developed to determine Cephalothin antibiotic drug. This method is based on the adsorptive accumulation of the drug at a hanging mercury drop electrode and then a negative sweep was initiated, which yield a well defined cathodic peak at -625 mV versus Ag/AgCl reference electrode. To achieve high sensitivity, various experimental and instrumental variables were investigated such as supporting electrolyte, pH, accumulation time and potential, drug concentration, scan rate, convection rate and working electrode area. The monitored adsorptive current was directly proportional to the concentration of Cephalothin and it shows a linear response in the range from 4 x 10(-7) to 1.2 x 10(-6) mol l(-1) (correlation coefficient=0.9995) and the detection limit (S/N=3) is 3.3 x 10(-9) mol l(-1) at an accumulation time of 3 min. The developed AdSV procedure shows a good reproducibility, the relative standard deviation R.S.D.% (n=10) at a concentration level of 5 x 10(-7) mol l(-1) was 0.94%. Possible interferences by other pharmaceutical drugs and surfactants have been also evaluated. The applicability of this approach was illustrated by the determination of Cephalothin in pharmaceutical preparation and biological fluids such as serum and urine. PMID- 15336348 TI - Development of methodologies based on HPLC and Raman spectroscopy for monitoring the stability of lovastatin in solid state in the presence of gallic acid. AB - Methodologies based on FT-Raman spectroscopy and HPLC were developed for monitoring the stability of lovastatin in the solid state in the presence of gallic acid, a natural antioxidant. A Raman calibration curve was constructed using the area of the strong but overlapping vibration mode of lovastatin at 1645 cm(-1) and of the gallic acid at 1595 cm(-1). Mixtures of the active ingredient with the antioxidant were heated in the presence of atmospheric air up to 120 degrees C. The molar ratios of lovastatin and gallic acid in the artificially oxidized mixtures were determined from their Raman spectra using the calibration curve. The results were compared to those obtained from the application of the HPLC methodology and found to match satisfactorily. The HPLC analysis was based on a reserved-phase Zorbax C(g), 10 microm (4.6mm x 25 cm i.d.) column, using a gradient elution program by varying the proportion of solvent A acetonitrile 100% to solvent B 0.1% v/v phosphoric acid, and a programmable diode array detection at 225 nm. The Raman methodology was simpler and non-destructive for the sample but yielded only molar ratios as opposed to the HPLC technique where the moles of the both ingredients were determined. PMID- 15336349 TI - A validated chiral LC method for the enantioselective analysis of Levetiracetam and its enantiomer R-alpha-ethyl-2-oxo-pyrrolidine acetamide on amylose-based stationary phase. AB - A new, simple chiral HPLC method was developed for the enantiomeric separation of Levetiracetam, [(S)-alpha-ethyl-2-oxo-pyrrolidine acetamide], an antiepileptic drug in pharmaceutical formulations and in bulk materials. Enantiomeric separation was achieved on a chiralpak AD-H column using a mobile phase consisting of hexane and isopropanol in the ratio (90:10, v/v) at a flow rate of 1.0 ml/min. The resolution between the enantiomers was found to be not less than 7 in the optimized method. Interestingly, unwanted enantiomer, namely R-alpha ethyl-2-oxo-pyrrolidine acetamide ((R)-enantiomer), was eluted prior to its mirror image in the developed method. The developed method was found to be selective in the presence of related impurities of Levetiracetam, namely N-(1 carbamoyl-propyl)-4-chloro-butyramide (Imp-1) and 1-ethyl-2-oxo-1-pyrrolidine acetic acid (Imp-2), and also under exposed conditions of UV light and 60 degrees C. The limit of detection (LOD) and limit of quantification (LOQ) of (R) enantiomer were found to be 900 and 2250 ng/ml, respectively, for 10 microl injection volume. The method precision for (R)-enantiomer at limit of quantification level was within 8% R.S.D. Calibration curve for (R)-enantiomer was linear over the studied ranges (2250-9000 ng) with correlation coefficient greater than 0998. The active pharmaceutical ingredient was extracted from its finished dosage form (tablet) using isopropanol. The percentage recoveries of (R) enantiomer were ranged from 94.2 to 102.6 and from 93.5 to 104.1 in spiked bulk and formulation samples of Levetiracetam, respectively. Levetiracetam sample solution and mobile phase are found to be stable for at least 48 h. The developed method was found to be rugged and robust. The proposed method was found to be suitable and accurate for the quantitative determination of (R)-enantiomer in bulk drugs and commercial formulations. Chiralcel OD-H column can also be used as an alternative column for the above purpose. PMID- 15336350 TI - Bead injection spectroscopy-flow injection analysis (BIS-FIA): an interesting tool applicable to pharmaceutical analysis. Determination of promethazine and trifluoperazine. AB - A bead injection spectroscopy-flow injection analysis (BIS-FIA) system for the spectrophotometric detection of promethazine and trifluoperazine is developed. The sensor is based in the oxidation of the phenothiazines by Fe(III) which is later determined by formation of the complex between Fe(II) and Ferrozine, [FeFz(3)](4-). Immediately, this complex is retained on a homogeneous bead suspension of Sephadex QAE A-25 resin (500 microl) which has been injected previously in the system to fill a commercial flow-cell (Hellma 138-OS). The use of BI with respect to the use of a reusable flow-through sensor is justified because the complex is so strongly retained on the beads that the regeneration of the solid support becomes extraordinarily difficult in the proposed method. At the end of the analysis, beads are automatically discarded from the flow-cell, by reversing the flow, and transported out of the system. The analytical signals are measured at a wavelength of 567 nm, corresponding to the absorbance of the complex. Using a sample volume of 600 microl, the analytical signal showed a very good linearity in the range 0.5-8.0 microgml(-1) and 0.5-10.0 microgml(-1), with detection limits of 0.09 and 0.14 microgml(-1) for promethazine and trifluoperazine, respectively. R.S.D.s (%) lower than 2% were obtained for both analytes. The proposed method is highly selective in the presence of other species that are normally encountered with these analytes. The sensor was satisfactorily applied to pharmaceutical preparations. PMID- 15336351 TI - LC/MS for the degradation profiling of cough-cold products under forced conditions. AB - Heat, acid, base, UV radiation and oxidation stress methods were applied to study the stability of cough-cold products containing acetaminophen, phenylephrine or phenylpropanolamine hydrochloride and chlorpheniramine maleate. Liquid chromatography coupled with mass spectrometry was used to analyze the degraded samples and obtain molecular weights information. Different volatile buffers (ammonium bicarbonate and ammonium acetate) were assayed in LC/MS methods and retention times of the analytes were compared with those obtained in HPLC with UV detection employing a conventional sodium phosphate buffer to establish the possibility of results transference between the two systems. PMID- 15336352 TI - Identification and characterization of potential impurities of donepezil. AB - Five unknown impurities ranging from 0.05 to 0.2% in donepezil were detected by a simple isocratic reversed-phase high performance liquid chromatography (HPLC). These impurities were isolated from crude sample of donepezil using isocratic reversed-phase preparative high performance liquid chromatography. Based on the spectral data (IR, NMR and MS), the structures of these impurities were characterised as 5,6-dimethoxy-2-(4-pyridylmethyl)-1-indanone (impurity I), 4 (5,6-dimethoxy-2,3-dihydro-1H-2-indenylmethyl) piperidine (impurity II), 2-(1 benzyl-4-piperdylmethyl)-5,6-dimethoxy-1-indanol (impurity III) 1-benzyl-4(5,6 dimethoxy-2,3-dihydro-1H-2-indenylmethyl) piperidine (impurity IV) and 1,1 dibenzyl-4(5,6-dimethoxy-1-oxo-2,3-dihydro-2H-2-indenylmethyl)hexahydropyridinium bromide (impurity V). The synthesis of these impurities and their formation was discussed. PMID- 15336353 TI - Analysis of low-molar-mass materials in commercial rubber samples by Soxhlet and headspace extractions followed by GC-MS analysis. AB - Very tight regulations apply to materials used for pharmaceutical packaging and for administering drugs. In this paper, we describe a simple and reliable procedure involving both gas- and liquid-phase extraction steps followed by an analysis step to identify the low-molar-mass materials in commercial-rubber samples. Representative commercial rubbers, that could be used for pharmaceutical packaging, have been selected and cryogenically powdered. Headspace and Soxhlet extractions have been carried out and the key parameters are discussed. The obtained extracts have been analyzed by gas chromatography (GC)-mass spectrometry (MS). More than 100 compounds have been detected and identified. Headspace allowed to extract the more-volatile compounds, whereas Soxhlet extraction recovered less-volatile compounds, but induced a loss of the volatile ones. Thus, both extraction techniques are required to fully characterize the low-molar-mass compounds present in rubber. PMID- 15336354 TI - Polarographic behavior of cephalexin and its determination in pharmaceuticals and human serum. AB - Cephalexin gives a reduction wave in 0.03 mol/l HCl medium at ca. -1.24 V. With cephalexin concentration higher than 2.5 x 10(-5) mol/l, another reduction wave is observed at ca. -0.90 V. These reduction waves are attributed to the reduction of ethylenic bond of a six-membered dihydrothiazine ring. When H2O2 is present, the reduction wave at ca. -0.90 V is catalyzed by H2O2 and its reduction intermediate hydroxyl radical *OH, producing a catalytic wave. However, the reduction wave at ca. -1.24 V remains nearly unchanged. A sensitive polarographic method for the determination of cephalexin is proposed based on the reduction wave of cephalexin. The second-order derivative peak current of the wave at ca. 1.24 V is rectilinear to the cephalexin concentration in the range 1.0 x 10(-7) to 2.5 x 10(-5) mol/l, and the detection limit is 5.0 x 10(-8) mol/l. The proposed method is applied to the individual tablet dosage form and human serum. PMID- 15336355 TI - Ruggedness/robustness evaluation and system suitability test on United States Pharmacopoeia XXVI assay ginsenosides in Asian and American ginseng by high performance liquid chromatography. AB - The work of the ruggedness/robustness evaluation and system suitability tests was oriented to profound understand the practicability of using assay methods issued by United States Pharmacopoeia (USP XXVI and XXVII) for ginsenosides in Asian ginseng and American ginseng. The items chosen for the method validation included quantitative related items such as recovery of Rg(1) and Rb(1), respectively, and qualitative related items such as resolution, theoretical plate number, relative retention time of two critical-band-pairs, Rg(1)/Re and Rb(1) with its neighboring peak, respectively. Totally, 16 column types were used for comparison of different vendors, different packing materials, different size, etc. and five sets of LC systems and two laboratories were involved in comparing the data of both quantitative and qualitative items. The results showed that different packing materials of columns used might significantly alters separation. The column packing material Hypersil afforded the preferable separating for the ginsenosides. No significant difference was observed from the different instrumentations and inter-laboratories. Our results suggest a modification of the system suitability test as given in USP26-NF21 and the latest version of USP27-NF22, which was not suitable for most systems. Using resolutions of Rg(1)/Re and Rb(1) with its neighboring peak as critical parameters for the ginsenosides assay and omitting the relative retention time of both Rg(1)/Re and Rb(1) with its neighboring peak is our suggestion for a more reasonable, yet practicable system suitability. Six typical chromatograms gain from different columns were figured out as well. PMID- 15336356 TI - Microdialysis for evaluating the entrapment and release of a lipophilic drug from nanoparticles. AB - The aim of the study was to evaluate the microdialysis (MD) as a tool to determine entrapment efficiency and drug release of a lipophilic drug model, diclofenac (DIC), from nanocapsules, nanospheres, and nanoemulsions. Factors that could interfere with the MD probe recovery were investigated: perfusion fluid composition, concentration and form of the drug in the perfusate, and recovery method. DIC entrapment efficiency to nanoparticles and the drug release in phosphate buffer pH 7.4 after different dilutions were evaluated by MD and ultrafiltration/centrifugation (UC). DIC recovery for the 5 microL/min flux was concentration and pH dependent. DIC sodium was used for the recoveries determination since it did not differ from the DIC acid recovery for the same media. DIC entrapment efficiency determined applying both techniques were equivalent and close to 100% for all nanoparticles. In pH 7.4 DIC release from the nanoparticles was partial for the dilution rate 1:1 (v/v), around 50-60%. A complete release was observed from 1:10 (v/v) dilution. Only nanocapsules presented a incomplete release for 1:5 (v/v) dilution, around 86%. MD and UC techniques were equivalent for the evaluation of DIC entrapment efficiency and drug release from the nanoparticles. PMID- 15336357 TI - Identification and chemical profiling of monacolins in red yeast rice using high performance liquid chromatography with photodiode array detector and mass spectrometry. AB - Monascus purpureus-fermented rice (red yeast rice) was one of the food supplements that had the ability of lowering the blood-lipid levels, and monacolins have been proved to be main active constituents. In total 14 monacolin compounds such as monacolin K (mevinolin), J, L, M, X, and their hydroxy acid form, as well as dehydromonacolin K, dihydromonacolin L, compactin, 3alpha hydroxy-3,5-dihydromonacolin L, etc. were identified in red yeast rice, using high-performance liquid chromatography with photodiode array detector and tandem mass spectrometry. A chemical fingerprint profiling method to display bioactive monacolins in red yeast rice was established and could be used for the quality control of the target material and its related products. Ten finish products labeled as red yeast rice from different manufacturers in marketing were traced using the chromatographic chemical profiling method, and the results show that only two of them were similar while the other eight were significantly different from the reference red yeast rice. All of these materials including raw material powder and finished products available were quantified and the contents of monacolins were calculated with reference of monacolin K (mevinolin) as the standard. PMID- 15336358 TI - Degradation of camptothecin-20(S)-glycinate ester prodrug under physiological conditions. AB - We have compared the strikingly different decomposition pathways for camptothecin 20(S)-acetate -acetate and camptothecin-20(S)-glycinate in phosphate buffered saline, human plasma and blood. The aliphatic ester analog camptothecin-20(S) acetate demonstrated excellent stability in the above fluids for many hours with minimal hydrolysis, while the camptothecin-20(S)-glycinate analog (differing solely by the presence of an amino group) underwent rapid and essentially complete decomposition. Reversed-phase high performance liquid chromatography (RP HPLC) with electrospray ionization-mass spectral (ESI-MS) detection was then used to correlate structural information for camptothecin-20(S)-glycinate decomposition products. ESI-MS detection indicated the ring-opened carboxylate form of camptothecin and the ring-opened degradation product co-elute near the solvent front, while the latest eluting decomposition product was the closed-ring lactone form of camptothecin. A novel decomposition product with intermediate retention time displayed an identical mass-to-charge ratio as camptothecin-20(S) glycinate ester but a strikingly different fragmentation pattern. The LC-ESI-MS evidence of a novel camptothecin prodrug degradation pathway is provided in this report. PMID- 15336359 TI - Quantitation of poloxamers in pharmaceutical formulations using size exclusion chromatography and colorimetric methods. AB - Poloxamers have been used as functional excipients in pharmaceutical products. They function as surfactants, emulsifying agents, solubilizing agents, dispersing agents, and in vivo absorbance enhancer. Despite their wide range of applications, limited analytical techniques have been reported in literature for characterizing poloxamers and few are targeted to quantify poloxamer contents in formulations with desired sensitivity and accuracy. In this paper, two distinct analytical methods for quantifying low levels of poloxamers in pharmaceutical formulations have been developed and optimized: a colorimetric method and a size exclusion chromatography method. The colorimetric method is based on the formation of a colored complex between poloxamers and cobalt(II) thiocyanate in aqueous medium, which has a maximum UV absorbance at 624 nm. The feasibility of this method is product specific. In this report, adequate specificity and sensitivity was demonstrated for only one of the several products tested. The size exclusion chromatography (SEC) method utilizes size exclusion columns with THF as mobile phase and refractive index detection. The SEC method provides a limit of quantitation (LOQ) of 0.005 mg/mL (0.0005%, w/w) and at least three orders of magnitudes of linear range. We applied the SEC method to pharmaceutical products containing 0.3-10% poloxamer 188 or poloxamer 407, such as Avapro, Neurontin, Sudafed and other developmental formulations. The results obtained with the SEC method agreed very well with literature and theoretical values with 97-102% recovery. The SEC method was proven to be widely applicable, accurate, precise and simple to use. PMID- 15336360 TI - High-performance liquid chromatographic analysis of a selective cyclooxygenase-1 inhibitor SC-560 in rat serum: application to pharmacokinetic studies. AB - A method of analysis of SC-560 (5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3 (trifluoromethyl)-1H-pyrazole) in biological fluids is necessary to study the kinetics of in vitro and in vivo metabolism. A simple high-performance liquid chromatographic method was developed for simultaneous determination of SC-560 and other products of metabolism in rat serum. Serum (0.1 ml) was precipitated with acetonitrile after addition of the internal standard, testosterone 17-propionate. Separation was achieved on a C(8) column with UV-detection at 240 nm. The calibration curve was linear ranging from 0.02 to 100 microg/ml. The mean recovery was >86.7%. Precision of the assay was <10% (R.S.D.%), and was within 15% at the limit of quantitation (20 ng/ml). Bias of the assay was lower than 15.5%. The limit of detection was 10 ng/ml for a 0.1 ml sample. The assay was applied successfully to the in vivo kinetic study of SC-560 in rats. PMID- 15336361 TI - A study of matrix effects on an LC/MS/MS assay for olanzapine and desmethyl olanzapine. AB - The purpose of this research project was to investigate potential matrix effects of anticoagulant and lipemia on the response of olanzapine, desmethyl olanzapine, olanzapine-D(3) and desmethyl olanzapine-D(8) in an LC/MS/MS assay. Blank human serum and sodium heparin, sodium citrate, and K(3)EDTA plasma with various degrees of lipemia were fortified with olanzapine, desmethyl olanzapine, olanzapine-D(3) and desmethyl olanzapine-D(8). Six replicates of each sample were extracted using Waters Oasis MCX cartridges and analyzed using electrospray LC/MS/MS. The analytes were separated on a Phenomenex LUNA phenyl hexyl, 2 mm x 50 mm, 5 microm, analytical column and a gradient rising from 2 to 85% mobile phase B. Mobile phase A consisted of acetonitrile-ammonium acetate (20 mM) (52:48 v/v) and mobile phase B was formic acid-acetonitrile (0.1:100 v/v). Ion suppression was investigated through post column infusion experiments. The degree of lipemia of each sample, indicated by turbidity, was ranked into categories from least to greatest and used for statistical analyses. The results from analysis of variance testing indicated that lipemia, anticoagulant and their interaction significantly influenced mass spectral matrix effects and extraction matrix effects. Differential behavior between the analytes and labeled internal standards contributed to variability. The most significant source of variability however, was ion suppression due to co-eluting matrix components. PMID- 15336362 TI - Evaluation of immunoassays for the measurement of erythropoietin (EPO) as an indirect biomarker of recombinant human EPO misuse in sport. AB - The measurement of serum erythropoietin (EPO) has been proposed as one of the indirect biomarkers for the detection of recombinant human EPO misuse in sport. An extended inter-laboratory validation of two commercial immunoassays for EPO measurement is described. A chemiluminescent immunoassay kit (CHEM) and an enzyme linked immunosorbent assay kit (ELISA) were evaluated. The CHEM assay showed intra-laboratory precision better than 6% and correct accuracy values for all quality control samples tested. Precisions and accuracies better than 7 and 13%, respectively, were obtained for the ELISA assay for most of the quality control samples. The limit of quantification estimated for CHEM assay was lower than for the ELISA assay. Inter-laboratory concordance was good for both the assays, with lower dispersion shown by the CHEM assay. Results obtained with the ELISA assay were always lower than those of the CHEM assay. However, a good inter-technique correlation was obtained ([ELISA]=0.76 [CHEM]+0.06, r2=0.92). Quality control samples had a good stability after one and two freeze/thaw cycles and in simulated transportation conditions. In conclusion, CHEM and ELISA assays showed similar characteristics regarding intra-laboratory validation. Better inter laboratory results were obtained with the CHEM assay and, hence, it is considered the recommended assay for anti-doping control analysis. PMID- 15336363 TI - Development of a chromatographic bioreactor based on immobilized beta glucuronidase on monolithic support for the determination of dextromethorphan and dextrorphan in human urine. AB - We here reported the development and application of an immobilized enzyme reactor (IMER) based on beta-glucuronidase to the on-line determination of urinary molar ratios of dextromethorphan (DOMe)/dextrorphan (DOH) for the assessment of the metabolic activity of CYP2D6, a genetically variable isoform of cytochrome P-450 (CYP). beta-Glucuronidase was immobilized on an HPLC monolithic aminopropyl silica support. Catalytic activity and stability of the chromatographic reactor were evaluated using 8-hydroxyquinoline glucuronide (8-HQG) as substrate. The IMER was coupled through a switching valve to a reversed-phase column (C8) for the simultaneous determination of dextromethorphan and its main metabolite dextrorphan. On purpose a selective reversed-phase ion pair HPLC method coupled with fluorescence detection was developed. Urine samples were first centrifuged to remove insoluble materials and then aliquots of the supernatants were injected into the coupled-column analyser. Linearity, precision and accuracy of the method were established. The method reliability was verified by comparing our data with previous data of a phenotyping study carried out by the Poison Control Centre of Pavia-Clinical Toxicology Division. PMID- 15336364 TI - Quantitative determination of albendazole metabolites in sheep spermatozoa and seminal plasma by liquid chromatographic analysis with fluorescence detection. AB - A new analytical method for the simultaneous quantitative determination of albendazole metabolites in sheep spermatozoa and seminal plasma at levels down to 46.5 ng/mL for albendazole sulphoxide (ABZ-SO), 7.5 ng/mL for albendazole sulphone (ABZ-SO2) and 12 ng/mL for albendazole 2-aminosulphone (ABZ-SO2NH2) has been developed. Analytes were extracted from alkalinized samples with ethyl acetate. Separation was carried out on a C18 column in the presence of tetra-n butylammonium (TBA) hydrogen sulphate and octanesulphonate sodium (OCT), as ion pair agents. Fluorometric detection was performed with excitation and emission wavelengths set at 290 and 320 nm, respectively. Accuracy data showed overall recoveries (+/-S.E.M.) of 83.1+/-1.2% for ABZ-SO, 98.8+/-0.6% for ABZ-SO2 and 85.3+/-0.7% for ABZ-SO2NH2, in spermatozoa. Respective values in seminal plasma were 88.0+/-0.9%, 97.7+/-0.5% and 93.2+/-1.7%. Precision data suggested coefficient of variation (CV%) values lower than 5.9% for spermatozoa and 3.8% for seminal plasma. The method was successfully applied for the determination of the three albendazole metabolites in semen samples collected from rams that had been orally administered albendazole. PMID- 15336365 TI - A simplified analytical method for a phenotyping cocktail of major CYP450 biotransformation routes. AB - An efficient, fast and reliable analytical method was developed for the simultaneous evaluation of the activities of five major human drug metabolising cytochrome P450 (1A2, 2C9, 2C19, 2D6 and 3A4) with a cocktail approach including five probe substances, namely caffeine, flurbiprofen, omeprazole, dextromethorphan and midazolam. All substances were administered simultaneously and a single plasma sample was obtained 2h after the administration. Plasma samples were handled by liquid-liquid extraction and analysed by gradient high performance liquid chromatography (HPLC) coupled to UV and fluorescence detectors. The chromatographic separation was achieved using a Discovery semi micro HS C18 HPLC column (5 microm particle size, 150 mm x 2.1 mm i.d.) protected by a guard column (5 microm particle size, 20 mm x 2.1 mm i.d.) The mobile phase was constituted of a methanol, acetonitrile and 20mM ammonium acetate (pH 4.5) with 0.1% triethylamine mixture and was delivered at a flow rate of 0.3 mL min( 1). All substances were separated simultaneously in a single run lasting less than 22 min. The HPLC method was formally validated and showed good performances in terms of linearity, sensitivity, precision and accuracy. Finally, the method was found suitable for the screening of these compounds in plasma samples. PMID- 15336366 TI - Simultaneous determination of Aprepitant and two metabolites in human plasma by high-performance liquid chromatography with tandem mass spectrometric detection. AB - A method for the simultaneous determination of Aprepitant, I (5-[[2(R)-[1(R)-(3,5 bistrifluoromethylphenyl)ethoxy]-3(S)-(4-fluorophenyl) morpholin-4-yl]methyl]-2,4 dihydro-[1,2,4]triazol-3-one) and two active metabolites (II and III) in human plasma has been developed. The method was based on high-performance liquid chromatography (HPLC) with atmospheric pressure chemical ionization tandem mass spectrometric (APCI-MS-MS) detection in positive ionization mode using a heated nebulizer interface. The analytes and internal standard (IV) (Fig. 1) were isolated from basified plasma using liquid-liquid extraction. The organic extracts were dried, reconstituted in mobile phase and injected into the HPLC MS/MS system. The analytes were chromatographed on a narrow bore (50 mm x 2.0 mm, 3 microm) Keystone Scientific's Prism R.P. analytical column, with mobile phase consisting of acetonitrile (ACN):water containing trifluoroacetic acid with pH adjusted to 3 (40:60, v/v) pumped at a flow rate of 0.5 ml/min. The MS-MS detection was performed on a Sciex API 3000 tandem mass spectrometer operated in selected reaction monitoring mode. The precursor-->product ion combinations of m/z 535-->277, 438-->180, 452-->223 and 503-->259 were used to quantify I, II, III, and IV, respectively, after chromatographic separation of the analytes. The assay was validated in the concentration range of 10-5000 ng/ml for I and II and 25-5000 ng/ml for III when 1 ml of plasma was processed. The precision of the assay (expressed as coefficient of variation, CV) was less than 10% at all concentrations within the standard curve range, with adequate assay accuracy. Matrix effect experiments were performed to demonstrate the absence of any significant change in ionization of the analytes when comparing neat standards to analytes in the presence of plasma matrix. This assay was utilized to support a clinical study where multiple oral doses of I were administered to healthy subjects to investigate the pharmacokinetics, safety, and tolerability of Aprepitant. Concentrations of the two most active metabolites, which if present in high concentrations would increase the neurokinin-1 (NK1) receptor occupancy level and therefore potentially contribute to the antiemetic action of Aprepitant, were determined. PMID- 15336367 TI - Comparison of extraction of a beta-blocker from plasma onto a molecularly imprinted polymer with liquid-liquid extraction and solid phase extraction methods. AB - An optimised solid phase extraction (SPE) method developed for the extraction of a structural analogue of the beta-blocking drug propranolol from plasma utilising a molecularly imprinted polymer (MIP) has been compared with methods based on conventional liquid-liquid extraction (LLE), and SPE using C18-bonded and immobilised phenyl boronic acid (PBA). All four methods could be used for the extraction of the analyte with acceptable accuracy and precision. The MIP-based method, unlike the other methods required a protein precipitation step prior to extraction to eliminate the effects of co-extracted protein. The best performance was seen with the LLE method followed by SPE on the C18 phase. The MIP-based method represented no advantage over the comparator methods for this analyte. Indeed the performance of the MIP-based method was marginally worse as leaching of low level template impurities prevented detection of the target analyte at low concentrations (5 ngmL(-1)). This relatively poorer performance was evident as worse accuracy at low concentrations with a consequent higher limit of quantification than the conventional methods. PMID- 15336368 TI - Conformational analysis: a tool for the elucidation of the antioxidant properties of ferulic acid derivatives in membrane models. AB - With the aim to search and design more effective and safe antioxidant molecules to be used as functional ingredients in cosmetic formulations for UV protection, we evaluated the antioxidant/radical scavenging activities of ferulic acid and of some alkyl ferulates in both acellular and cellular systems. Ferulic acid esters, equipotent as antioxidant in homogeneous phase, showed when tested in membranous systems (rat liver microsomes, rat erythrocytes) marked differences in antioxidant potency. The n-C(12) derivative was the most potent, followed by n C(8), n-C(16) and branched C(8), and then by ferulic acid. A conformational study carried out by NMR and modelling, indicates that the different antioxidant activity of ferulates in membrane models is due to the different spatial conformation and arrangement of the side chain of the molecule, which governs the access and binding to the phospholipid bilayer, the modality of orientation of the scavenging/quenching nucleus (phenol moiety), and hence the overall antioxidant potency of the derivative. These results emphasize the need of analytical studies (NMR and molecular modelling) addressed to the knowledge of the conformational parameters in combination with conventional antioxidant testings for understanding the antioxidant behaviour of a molecule in a biological membrane/system. PMID- 15336369 TI - Shelf lives of aseptically prepared medicines--stability of netilmicin injection in polypropylene syringes. AB - There is no published information on the stability of netilmicin solutions in prefilled syringes. The purpose of this study was to evaluate the stability of netilmicin in polypropylene syringes and to determine the optimum validated shelf life so that they may be prepared in bulk in appropriately licensed facilities. The syringes containing netilmicin 10 or 100mg/ml were stored at 7 degrees C, room temperature in the light (RTL) and 25 degrees C/60% relative humidity for up to 300 days. Netilmicin concentration was determined by reversed phase high performance liquid chromatography (RP-HPLC) of the isoindole derivative formed with o-phthalaldehyde (OPA). The shelf lives were calculated using the maximum rate method applied to the netilmicin analytical data. At 7 degrees C 10 and 100mg/ml solutions were stable for 90 days falling to 30 days at 25 degrees C and 60% RH. At RTL the 10mg/ml solution was stable for 9 days. PMID- 15336370 TI - Determination of the ionization constants of 4-iodo-2,6 dimethylphenylcarbamoylmethyl iminodiacetic acid. AB - In order to develop a radiopharmaceutical for hepatobiliary scintigraphy with better hepatobiliary properties new ligand for complexation of 99mTc, 4-iodo-2,6 dimethylphenylcarbamoylmethyl iminodiacetic acid (METHYLIODIDA), was synthesized. Acid-base equilibria of METHYLIODIDA were studied potentiometrically, because these data are important for determination of complex formation conditions. It was established that METHYLIODIDA undergoes a complex acid-base equilibrium due to its zwitterionic nature and four proton-binding sites. The stoichiometric ionization constants were determined at 25 degrees C and constant ionic strength 0.1M (NaClO(4)): pK1=1.7+/-0.1; pK2=2.44+/-0.07; pK3=6.29+/-0.02 and pK4=10.91+/ 0.06, respectively. PMID- 15336371 TI - LC-MS/MS and FT-IR analyses of stones from a patient with Crohn's disease: a case report. AB - This report describes the unusual case of a patient affected by Crohn's disease suffering from intestinal obstruction with recurrent occlusive symptoms not due to the intestinal disease, but to the presence of two calcified foreign bodies in the pelvis. The stones were surgically removed and analysed by reverse-phase liquid chromatography coupled to UV diode array detection and mass spectrometry (LC-UV-DAD-MS/MS), Chromatoprobe-MS/MS and by Fourier-transform-infrared spectroscopy (FT-IR) techniques. The combined mass spectrometric approaches allowed unequivocally to identify 5-aminosalicylic acid (5-ASA) in stone 1, and to demonstrate that its formation was due to an unmodified 5-ASA tablet, a formulation that must undergo complete dissolution in the small bowel. The second stone was constituted by a solid layer (no solvent-extractable material) identified by FT-IR as a polystyrene fragment. This indicates that accidental ingestion of a plastic material, followed by its calcification, was responsible for its formation. PMID- 15336372 TI - The influence of pH, temperature and buffers on the degradation kinetics of cefetamet pivoxil hydrochloride in aqueous solutions. AB - The first-order hydrolysis kinetics of cefetamet pivoxil (CP) were investigated as a function of pH, temperature and buffers. The degradation was followed by HPLC. Buffer catalysis was observed in acetate and phosphate buffers. The pH-rate profiles for hydrolysis of cefetamet pivoxil were obtained at 333, 343, 353 and 363K. The pH-rate expression was k(pH)=kH+aH+ + kH2OkOH-aOH-, where kH+ and kOH- are the second-order rate constants (mol(-1)ls(-1)) for hydrogen ion activity and for hydroxyl ion activity respectively, and kH2O is the pseudo-first-order rate constant (s(-1)) for spontaneous reaction under the influence of water. The pH rate profile was characteristically U-shaped. Maximum stability was observed in the pH region from 3 to 5. PMID- 15336373 TI - A validated chiral HPLC method for the enantiomeric separation of tolterodine tartarate. AB - An isocratic chiral HPLC method was developed for the separation of tolterodine tartarate enantiomers. The mobile phase consists of n-hexane and isopropyl alcohol in the ratio of 980:20 (v/v) with 1 ml diethylamine and 0.6 ml trifluoroacetic acid. Chiralcel OD-H (250 mm x 4.6mm) column was used at constant room temperature. Flow rate was kept at 0.5 ml/min. This method is capable of detecting the S-isomer up to 0.1 microg/ml. The method was validated in terms of linearity, precision, limit of detection (LOD) and limit of quantification (LOQ). PMID- 15336374 TI - Simultaneous determination of roxithromycin and ambroxol hydrochloride in a new tablet formulation by liquid chromatography. AB - A rapid and accurate liquid chromatographic method is described for the simultaneous determination of roxithromycin and ambroxol hydrochloride in a new tablet formulation. Chromatographic separation of the two drugs was achieved on a Diamonsil C(18) column (200 mm x 4.6 mm, 5 microm). The mobile phase consisting of a mixture of acetonitrile, methanol and 0.5% ammonium acetate (39:11:50 (v/v), pH 5.5) was delivered at a flow rate of 1.0 ml/min. Detection was performed at 220 nm. Linearity, accuracy and precision were found to be acceptable over the concentration range of 201.2-2012.0 microg/ml for roxithromycin and 42.7-427.0 microg/ml for ambroxol hydrochloride, respectively. Separation was complete in less than 10 min. The proposed method can be used for the quality control of formulation products. PMID- 15336375 TI - Binding study of drug with bovine serum album using a combined technique of microdialysis with flow-injection chemiluminescent detection. AB - Microdialysis coupled with flow-injection chemiluminescence (FI-CL) has been developed to determine the binding parameters of a drug binding to protein by using antibiotic tetracycline hydrochloride binding to bovine serum albumin as a model system. The drug and protein were mixed in different molar ratios in 0.067 M phosphate buffer, pH 7.4, and incubated at 37 degrees C in a water bath. The microdialysis probe was utilized to sample the mixed solution at a perfusion rate of 5 microL/min. The concentration of unbound tetracycline hydrochloride in the microdialysate was determined by FIA-CL. In vitro recovery of tetracycline hydrochloride under experimental conditions was 30.0%. The data obtained by the present microdialysis-FI-CL system was analyzed using the Scatchard analysis and Klotz plot. The results show that the Scatchard plot and Klotz plot are linear with good correlation coefficient, indicating a good agreement of the experimental data and to the theoretical equation. The FIA chemiluminescence system combined with microdialysis developed in this work demonstrated its use for determination of interaction between drug and protein by using relatively simple instrument. PMID- 15336376 TI - Enzyme-amplified lanthanide luminescence based on complexation reaction--a new technique for the determination of doxycycline. AB - A new spectrofluorimetric method is described for the determination of doxycycline, based on modified enzyme-amplified lanthanide luminescence. Under the optimum conditions, Eu(3+)-doxycycline forms a ternary complex with lysozyme in close proximity and lysozyme can remarkably enhance the characteristic fluorescence intensity of Eu(3+) at 612nm in doxycycline-Eu(3+) binary complex. The enhanced fluorescence intensity is in proportion to the concentration of doxycycline. The limit of detection is 1.28 x 10(-8) moll(-1), with a linear range from 1.7 x 10(-7) to 1.7 x 10(-6) moll(-1). Interferences of other coexisting substances were studied. The developed method was successfully applied to the determination of doxycycline in serum, urine and real samples. The mechanism of fluorescence enhancement was also studied. PMID- 15336377 TI - Simultaneous determination of serum flecainide and its metabolites by using high performance liquid chromatography. AB - Simultaneous determination of serum flecainide and its oxidative metabolites was carried out by using high performance liquid chromatography (HPLC) equipped with conventional octadecylsilyl silica (ODS) column and fluorescence detector. Flecainide and its metabolites, m-O-dealkylated flecainide (MODF) and m-O dealkylated lactam of flecainide (MODLF) in serum were extracted with ethyl acetate. The recoveries of flecainide, MODF and MODLF were greater than 92, 93, and 60% with the coefficient of variations (CVs) less than 3.2, 5.8, and 5.3%, respectively. The calibration curves were linear at the concentration range of 50 1500 ng/mL for flecainide and 10-500 ng/mL for MODF and MODLF (r > 0.999). The CVs for intra-day assay were 2.7-5.3% for flecainide, 3.0-4.2% for MODF, and 3.7 4.3% for MODLF, respectively. The CVs for inter-day assay were 7.0-8.4% for flecainide, 3.3-6.7% for MODF, and 4.4-7.7% for MODLF, respectively. This assay method can be used for assessing the metabolic ability of flecainide in the patients with tachyarrhythmia. PMID- 15336378 TI - Polymer microspheres for controlled drug release. AB - Polymer microspheres can be employed to deliver medication in a rate-controlled and sometimes targeted manner. Medication is released from a microsphere by drug leaching from the polymer or by degradation of the polymer matrix. Since the rate of drug release is controlled by these two factors, it is important to understand the physical and chemical properties of the releasing medium. This review presents the methods used in the preparation of microspheres from monomers or from linear polymers and discusses the physio-chemical properties that affect the formation, structure, and morphology of the spheres. Topics including the effects of molecular weight, blended spheres, crystallinity, drug distribution, porosity, and sphere size are discussed in relation to the characteristics of the release process. Added control over release profiles can be obtained by the employment of core-shell systems and pH-sensitive spheres; the enhancements presented by such systems are discussed through literature examples. PMID- 15336379 TI - The effects of molecular weight and porosity on the degradation and drug release from polyglycolide. AB - This paper explores the influence of initial molecular weight and porosity on the release of the drug, theophylline, from polyglycolide (PGA). PGA was made by a variety of processes to vary the molecular weight and was blended with NaCl with different crystal sizes and in different proportions to vary the pore size and volume. Overall, results showed that decreasing the molecular weight and increasing the pore size and volume increased the rate of drug release. The exact variation of these trends agreed well with the previously established four-stage degradation mechanism for PGA, but was more complex than a simple linear behaviour. Because both the molecular weight and the porosity of PGA have a substantial influence on the polymer degradation, and can be varied in a controlled manner, these parameters can play an important role in developing PGA as a controlled drug delivery material with tailored drug release. PMID- 15336380 TI - Control, communication and monitoring of intravaginal drug delivery in dairy cows. AB - We present the design of an electronically controlled drug delivery system. The intravaginally located device is a low-invasive platform that can measure and react inside the cow vagina while providing external control and monitoring ability. The electronics manufactured from off the shelf components occupies 16 mL of a Theratron syringe. A microcontroller reads and logs sensor data and controls a gascell. The generated gas pressure propels the syringe piston and releases the formulation. A two way radio link allows communication between other devices or a base station. Proof of principle experiments confirm variable-rate, arbitrary profile drug delivery qualified by internal sensors. A total volume of 30 mL was dispensed over a 7-day-period with a volume error of +/- 1 mL or +/- 7% for larger volumes. Delivery was controlled or overridden via the wireless link, and proximity to other devices was detected and recorded. The results suggest that temperature and activity sensing or social grouping determined via proximity can be used to detect oestrus and trigger appropriate responses. PMID- 15336381 TI - Bioadhesive properties and rheology of polyether-modified poly(acrylic acid) hydrogels. AB - Transient rheological properties and mucoadhesion of hydrogels composed of poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) (PEO-PPO-PEO, or Pluronic) block copolymers and poly(acrylic acid) were explored. Nine Pluronic copolymers ranging in nominal molecular weight and PPO/PEO content were grafted to PAA through C-C bonds, with or without the use of divinyl cross-linker, ethylene glycol dimethacrylate (EGDMA). The hydrogel elasticity increased with the PPO content in the copolymers, as well as in the presence of EGDMA. Tensile tests were conducted to measure the fracture strength and the work of adhesion between the hydrogels and rat intestinal tissue. The fracture strength was proportional to the gel pseudoequilibrium modulus and depended on the nominal length of the PPO segments in the parent Pluronic copolymer. The work of mucoadhesion and gel cohesion declined with the loss angle measured in oscillatory shear experiments. The length of the PEO segments in Pluronic affected the work of adhesion. Applications of the Pluronic-PAA gels as vehicles in oral drug delivery are discussed. The longest Pluronic copolymers bonded to PAA resulted in copolymeric gels with strongest mucoadhesive properties. PMID- 15336382 TI - Paclitaxel-loaded poly(L-lactic acid) microspheres 3: blending low and high molecular weight polymers to control morphology and drug release. AB - Microspheres were prepared from paclitaxel and binary polymer blends incorporating 1, 3, 40k and 100k g/mol PLLA. Thermal analysis was performed by DSC and in vitro paclitaxel release profiles were determined at 37 degrees C in phosphate buffer using an HPLC assay. In microspheres made with 3k/40k PLLA blends, the glass transition (Tg), crystallinity and melting temperature (Tm) all decreased with an increasing proportion of low molecular weight polymer in the blend. Similar trends were observed for 1k/100k blends. Tm values ranged from 175 to 110 degrees C and Tg values between 66 and 37 degrees C. However, for 1k/100k blends, melting point depression was linearly dependent on blend composition when plotted as 1/Tm = 0.000109 x (%1k in blend) + 0.0223, R2 = 0.97. A similar plot with data from the 3k/40k system yielded a non-linear relationship. Furthermore, the decrease in Tg for both 1k/100k and 3k/40k blends followed the Fox equation, although experimental values were consistently 1-2 degrees C above predicted values. Paclitaxel release from microspheres made with a 1k/100k blend occurred in four distinct phases: a burst phase (day 0), a slower phase, a second burst (day 35) and a second slower phase (until day 70). The second burst coincided with visible degradation of the microspheres. Blends of low and high molecular weight PLLA display thermal properties indicating that 1k g/mol PLLA behaves as a diluent when blended with 100k g/mol PLLA, being excluded from the crystalline domains in the polymer matrix. In contrast, 3k g/mol PLLA is incorporated in both amorphous and crystalline regions of the polymer blend. Paclitaxel release profiles from 1k/100k PLLA microspheres demonstrate a multiphase profile due to the effects of both diffusion and degradation controlled release mechanisms. PMID- 15336383 TI - Solubility, dissolution rate and phase transition studies of ranitidine hydrochloride tautomeric forms. AB - Understanding the polymorphic behavior of pharmaceutical solids during the crystallization process and further in post-processing units is crucial to meet medical and legal requirements. In this study, an analytical technique was developed for determining the composition of two solid forms of ranitidine hydrochloride using two peaks of Fourier transform infrared (FTIR) spectra without the need to grind the samples. Solubility studies of ranitidine hydrochloride showed that Form 2 has a higher solubility than Form 1. Solution mediated transformation is very slow and occurs from Form 2 to Form 1 and not the reverse. No solid-solid transformation was observed due to grinding or compressing the pure samples of either forms and of a 50/50 wt.% mixture. Grinding was found to be a proper technique for increasing the bulk solid density of the ranitidine hydrochloride without the risk of solid-solid transformation. Dissolution rate found to be equally fast for both forms. The solubility data were modeled using the group contribution parameters and UNIversal QUAsi-Chemical (UNIQUAC) theory. There was a good agreement between the experimental solubility data of ranitidine hydrochloride and the results of UNIQUAC equation. PMID- 15336384 TI - Pharmacokinetics of a cephalone (CQ-M-EPCA) in rats after oral, intraduodenal and intravenous administration. AB - As part of the development of a new series of antibacterial agents derived from coupling a beta-lactamic precursor with a fluoroquinolone and named cephalones, the pharmacokinetics of one derivate: CQ-M-EPCA in rats after intravenous, intragastric and intraduodenal routes, was carried out. After the IV injection of 20 mg/kg or 40 mg/kg of this cephalone, plasma concentrations at the time zero (Cp0) were 3.1 and 11.26 microg/ml, respectively. Plasma concentrations decreased rapidly to almost disappear in both instances. Forty-five minutes later, a surge in concentrations, in the 40 mg/kg group, with a maximal plasma concentration (Cpmax) of 2.97 microg/ml was observed. An elimination half-life (T1/2el) of 2.36 +/- 0.33 h. was calculated. The drug was undetected by the ninth hour. Intragastric administration of the drug resulted in Cpmax of 3.78 +/- 0.26 microg/ml with a time to reach Cpmax (Tmax) of 25 min and T1/2el = 3.22 h. Same variables after intraduodenal administration were Cpmax 4.71 microg/ml; Tmax 1h, and T1/2el 3.41 h. Outstandingly high bioavailabilities after intragastric and intraduodenal administration (169 and 246%, respectively), together with the shape of the concentration versus time profiles after IV administration suggest that the drug undergoes a complex redistribution phenomenon, while showing high tissue diffusion with an apparent volume of distribution of 3.33 l/kg. PMID- 15336385 TI - Influence of human nail etching for the assessment of topical onychomycosis therapies. AB - The purpose of this investigation was to study the physico-chemical properties of hot-melt extruded films containing ketoconazole and to determine the influence of 'nail etching' on film bioadhesion and drug permeability for the assessment of topical onychomycosis therapies. Hot-melt extrusion (HME) was used to prepare films containing 20% w/w ketoconazole. Ketoconazole 0.125% gel was also prepared using Carbopol 974P NF. Films were processed at a temperature range of 115-120 degrees C utilizing a Killion extruder (KLB-100), and were evaluated for post extrusion drug content, content uniformity, bioadhesion, thermal behavior and nail drug permeation. The extruded films demonstrated excellent content uniformity and post-processing drug content. Tensile and peel tests were recorded to determine the bioadhesive profiles. In this study, work of adhesion and peak adhesive force determinations using the peel tests provided more sensitive results for evaluating the bioadhesivity of the HME films than the tensile tests. The in vitro permeability profiles have demonstrated, that nail samples treated with an 'etchant' demonstrated a significant increase in drug permeability compared to control. Differential scanning calorimetry (DSC) thermograms indicated that ketoconazole was in solid solution within the HME films. These findings are encouraging for the future design and formulation of novel drug delivery systems for the topical treatment of onychomycosis. PMID- 15336386 TI - Effect of particle morphology on the triboelectrification in dry powder inhalers. AB - Electrostatic charge of lactoses of different particle morphology and amorphous contents were measured during actuation from two different dry powder inhalers (DPIs). Triboelectric studies may give important information when new inhaler devices, materials and formulations are designed in order to improve the drug deposition. Two inhalers, Taifun (Focus Inhalation Oy, Finland) and Clickhaler (Innovata Biomed Ltd., UK) were filled with lactose powders which were spray dried from different solutions or suspensions of lactose, ethanol and water. Differences in the amorphous contents were determined with isothermal microcalorimetry (IMC) and X-ray diffraction (XRD) and the particle morphology was examined with laser diffraction and electron microscopy (SEM). Samples were actuated from the inhalers at given intervals into the Faraday pail and the generated charges were recorded. Increase in the water concentration of the feed suspension had negligible effect on the charging until it exceeded 70%. Reproducibility of the measurement was found to be better with samples of homogenous particles and higher crystallinity while more amorphous samples with different morphology and wide particle size distribution showed change in the sign of the charge in addition to higher variations of the magnitude. In this study we show that different inhalers, as well as the morphology of the lactose powder, has noticeable effects on the generated charge which has previously been shown to affect the deposition of the drug and the function of an inhaler. PMID- 15336387 TI - Rheological, mucoadhesive and release properties of Carbopol gels in hydrophilic cosolvents. AB - Carbopol is one of the most common thickening agent for water phases. It is used after neutralisation and its rheological properties in the aqueous medium are well known. The aim of this work was to investigate the gelation properties of Carbopol 971 e 974 polymeric systems in water-miscible cosolvents such as glycerine and PEG 400. Since in these cosolvents, carboxypolymethylene precipitates after neutralisation with a base, then the attention was pointed out of the gelation properties of the different systems at increasing temperature, in order to obtain Carbopols gels avoiding neutralisation and, at the same time, making possible the dissolution in these gels of insoluble or poorly soluble water drugs. Rheological properties of PEG 400 and glycerine samples were compared with similar systems in water by performing oscillatory analyses and measuring the main rheological parameters, G', G" and delta. The results obtained showed that Carbopol 971 and 974 in PEG 400 gave rise after heating to gels that show a satisfactory rheological behaviour. The elastic modulus is greater than the viscous one showing a remarkable elastic character of these samples and the performed frequency sweeps show a typical spectrum of a "gel-like" structure. Being Carbopols well-known mucoadhesive polymers, gels adhesive properties were studied using the ex vivo method. Then, the possible cutaneous irritation were also tested using the in vivo method (Draize test). No signs of cutaneous irritation and good mucoadhesive properties were obtained for the PEG 400 and water gels of Carbopol 974 prepared by heating. After rheological and mucoadhesive properties were set, paracetamol as a model drug was then inserted in the composition of the gels and the release characteristics were defined. Dissolution tests pointed out the greater release control properties of PEG 400 Carbopol 971 samples. These studies showed PEG 400-Carbopol systems as a first rate alternative to traditional water gels. PMID- 15336388 TI - In vivo evaluation of a reverse water-in-fluorocarbon emulsion stabilized with a semifluorinated amphiphile as a drug delivery system through the pulmonary route. AB - The potential of a reverse water-in-fluorocarbon (w-in-FC) emulsion stabilized with a semifluorinated amphiphile, namely C8F17(CH2)11OP(O)[N(CH2CH2)2O]2 (F8H11DMP) for drug delivery through intrapulmonary administration was investigated in the mouse. This study involved assessment of the effect of single or repeated intranasal instillations of a plain emulsion on lung tissue integrity, and evaluation of blood glucose levels in mice treated with an insulin loaded emulsion. When instilled intranasally to mice, the plain emulsion did not alter lung tissue integrity, as demonstrated by histological staining, and did not induce any airway inflammatory reaction. Treated mice exhibited decreased body weight within the 3-4 days that followed the first emulsion administration, but this decrease was reversible within few days. Mice instilled intranasally with the insulin-loaded emulsion displayed decreased blood glucose levels within the 20 min that followed the administration, thus demonstrating the potential of the reverse w-in-FC emulsion stabilized with F8H11DMP to systemically deliver drugs, including peptides, upon lung administration. PMID- 15336389 TI - Control of pulmonary absorption of water-soluble compounds by various viscous vehicles. AB - Effects of various viscous vehicles on the pulmonary absorption of water-soluble drugs were examined by an in situ pulmonary absorption experiment. Gelatin, polyvinylacohol (PVA), hydroxypropylcellose (HPC), chondroitin sulfate A sodium salt (CS), polyacrylic acid (PAA), methylcellulose #400 (MC400) and hyaluronic acid sodium salt (HA) were used as models of viscous vehicles. 5(6) Carboxyfluorescein (CF) and fluorescein isothiocayanate-labeled dextran with an average molecular weight of 4000 (FD4) were used as water-soluble drugs. The plasma concentration of CF was controlled and regulated in the presence of these viscous vehicles, especially gelatin (1-5%) and polyvinyl alcohol (PVA) 1%. In the pharmacokinetic analysis, the Cmax values of CF significantly decreased, and its Tmax values increased in the presence of these viscous vehicles compared with the control. The MRT and MAT values of CF with these vehicles were significantly higher than those without these vehicles. Therefore, these findings indicated that the viscous vehicles were effective to regulate the absorption rate of CF. On the other hand, the pulmonary absorption of FD4 was not so much affected even in the presence of gelatin and PVA, although PVA slightly decreased MRT value, and significantly decreased Tmax value. Furthermore, we examined the release rate of CF from the cellulose tube containing various concentrations of gelatin. The release rate of CF from the cellulose tube with gelatin was inversely related to the viscosity of gelatin. In addition, the release rate of CF was inversely related to DeltaMAT (DeltaMAT = MATgel(MAT with gelatin)-MATsol(MAT without gelatin)) in the presence of varying concentrations of gelatin. These findings indicated that these viscous vehicles were effective to control the pulmonary absorption of CF, a water-soluble drug with low molecular weight and they might be useful to increase the local concentration of drugs in the lung. PMID- 15336390 TI - Stability study of amorphous valdecoxib. AB - Formulation of poorly water-soluble drugs in the most stable dosage form for oral delivery perhaps presents the greatest challenge to pharmaceutical industry. Physical transformation of drug substance into its more soluble but metastable amorphous form is one of the approaches for improving dissolution rate of such drugs. The present study utilizes technique of spray drying for preparation of solid dispersions (SDs) and includes stability study of the same. Valdecoxib (VLD), a prototype of poorly water-soluble drugs, has been the drug of choice. The hydrophilic carriers selected were polyvinylpyrrolidone K30 (PVP) and hydroxypropylcellulose (HPC). SDs and pure VLD in the form of spray dried powder (SDVLD) in comparison with pure drug and corresponding physical mixtures (PMs) were initially characterized and then subjected to stability testing at ambient temperature and relative humidity up to 3 months. During initial characterization, increase in saturation solubility and dissolution rate was observed in all samples. DSC and XRPD studies of SDVLD and SDs suggested generation of amorphous form of drug. IR spectroscopy revealed presence of hydrogen bonding in SDs. During stability testing, there was gradual decrease in saturation solubility and dissolution rate of SDs, over the period of 3 months. While, saturation solubility of SDVLD dropped drastically within 15 days and was almost comparable with pure VLD. SD PVP retained the amorphous form of drug throughout stability period, whereas SD HPC and SDVLD presented incidence of crystallinity after 1 month and 15 days, respectively. This was justified by enthalpy relaxation studies in which, amorphous VLD showed considerable relaxation of enthalpy at Tg, while it was totally suppressed in SD PVP and partly in SD HPC. The study thus definitely reveals tremendous potential of solid dispersions of valdecoxib with PVP, from stability point of view. PMID- 15336391 TI - Flow microcalorimetric studies of phenol and its chlorinated derivatives and a theoretical evaluation of their possible inhibition mode on Chromobacterium violaceum respiration. AB - The general belief that chemical structure determines the biological effect of drugs has led to several techniques to establish structure-activity relationships (SAR) that is useful in the development of more active compounds. Predicting toxic effects based on SAR, one can obtain toxicological data with a low cost benefit ratio. Chlorophenols that represent a class of toxic agents frequently used in industrial processes are not satisfactorily described in the literature in relation to their toxicity. The main objective of this work is to relate the microbial activities of phenol, anisole and their chlorinated derivatives on Chromobacterium violaceum respiration with their physicochemical properties. Anisole and its chlorinated derivatives were used to evaluate the influence of phenol acidity on biological activity. The calculations were carried out at the semi-empirical AM1 and ab initio DFT levels employing the basis sets CEP-31G, CEP 31+Ge CEP-31G** that were parameterized using the continuum-solvation model COSMO for solvent contribution. Both empirical and theoretical properties were evaluated by chemometric analyses (hierarchical cluster analysis (HCA) and principal component analysis (PCA)), to correlate the physicochemical properties of the phenol, anisole and their chlorinated derivatives with their biological activities. The results obtained for the current work indicate that the biological activities of these compounds increase as the n-octanol/water (logP) partition coefficients, ionization energies (IP), melting points (mp) and dissociation constants increase and the solvent effects (SE), enthalpies of formation (DeltafH degrees ) and proton affinities (PA) decrease. PMID- 15336392 TI - Effects of intrinsic variables on release of sodium dodecyl sulfate from a female controlled drug delivery system. AB - The release profile of sodium dodecyl sulfate (SDS), a potent microbicide, from a female controlled drug delivery system (FcDDS) made of Carbopol 934P and hydroxypropyl methylcellulose (HPMC) was evaluated using a newly developed in vitro Simulant Vaginal System (SVS). The major parameters involved in the release profiles of SDS were categorized as: (1) formulation variables (total loading weight of intravaginal delivery systems, SDS loading doses in intravaginal delivery systems); (2) intrinsic variables (vaginal fluid secretion rate, vaginal fluid pH); and (3) extrinsic variables (inserting position). In most conditions, about 70% of the loading dose of SDS was released from FcDDS within 6h of application. The release profile showed that concentrations needed for complete human papilloma virus (HPV) inactivation could be obtained within 10 min after the application. It was demonstrated that intrinsic variables (i.e., the rate and pH of vaginal fluid) played an integral role in determining the release profile of SDS, while loading dose of SDS in FcDDS did not significantly affect the percentage of the total amount of SDS released. It can be concluded that FcDDS can be exploited as a controlled delivery device for prevention against sexually transmitted diseases. PMID- 15336393 TI - A flow injection analysis/mass spectrometry method for the quantification of polyethylene glycol 300 in drug formulations. AB - A direct flow injection analysis/mass spectrometry (FIA/MS) method was developed for the quantification of polyethylene glycol. The method was used for the evaluation of distribution uniformity and mixing homogeneity of polyethylene glycol 300 (PEG 300) as a component in drug formulation mixtures. In the method, five of the most intense ions of the PEG 300 oligomer were chosen for selected ion monitoring (SIM) by mass spectrometry. Standard calibration curves were established, using either single channel SIM or the summed intensity of all five SIM channels plotting against the standard concentrations. Both calibration approaches produced comparable results on quantification. The feasibility of the method was demonstrated using both atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI). The method provided fast and sensitive quantification of PEG 300 without tedious chromatographic separation or sample preparation. The method has been successfully adopted for the evaluation of the mixing process in drug formulations. PMID- 15336394 TI - Dynamic contact angle measurement on materials with an unknown wet perimeter. AB - Whilst contact angle measurements obtained using the Wilhelmy balance technique are accurate and reproducible for planar surfaces, their use for characterizing particulate materials is highly dependent upon accurate knowledge of the wet perimeter. This communication suggests that the approach of Pepin et al. [Int. J. Pharm. 152 (1997) 1] for wet perimeter determination using non-polar liquids may lead to erroneous conclusions. Alternative approaches for wet perimeter determination are suggested. PMID- 15336395 TI - Insertional mutagenesis and characterization of a polyketide synthase gene (PKS1) required for melanin biosynthesis in Bipolaris oryzae. AB - A polyketide synthase gene named PKS1, involved in the melanin biosynthesis pathway of the phytopathogenic fungus Bipolaris oryzae, was isolated using restriction enzyme-mediated integration. Sequence analysis showed that the PKS1 encodes a putative protein that has 2155 amino acids and significant similarity to other fungal polyketide synthases. Targeted disruption of the PKS1 gene showed that it is necessary for melanin biosynthesis in B. oryzae. Northern blot analysis showed that PKS1 transcripts were specifically enhanced by near ultraviolet radiation (300-400 nm) and that its temporal transcriptional patterns were similar to those of THR1 and SCD1 genes involved in the melanin biosynthesis pathway of B. oryzae. PMID- 15336396 TI - Identification, characterisation and specificity of a cell wall lytic enzyme from Lactobacillus fermentum BR11. AB - Screening of a genomic library with an antiserum raised against whole Lactobacillus fermentum BR11 cells identified a clone expressing an immunoreactive 37-kDa protein. Analysis of the 3010-bp DNA insert contained within the clone revealed four open reading frames (ORFs). One ORF encodes LysA, a 303 amino acid protein which has up to 35% identity with putative endolysins from prophages Lj928 and Lj965 from Lactobacillus johnsonii and Lp1 and Lp2 from Lactobacillus plantarum as well as with the endolysin of Lactobacillus gasseri bacteriophage Phiadh. The immunoreactive protein was shown to be encoded by a truncated ORF downstream of lysA which has similarity to glutamyl-tRNA synthetases. The N-terminus of LysA has sequence similarity with N acetylmuramidase catalytic domains while the C-terminus has sequence similarity with putative cell envelope binding bacterial SH3b domains. C-terminal bacterial SH3b domains were identified in the majority of Lactobacillus bacteriophage endolysins. LysA was expressed in Escherichia coli and unusually was found to have a broad bacteriolytic activity range with activity against a number of different Lactobacillus species and against Lactococcus lactis, streptococci and Staphylococcus aureus. It was found that LysA is 2 and 8000 times more active against L. fermentum than L. lactis and Streptococcus pyogenes, respectively. PMID- 15336397 TI - The interaction of Vatairea macrocarca and Rhizobium tropici: net H+ efflux stimulus and alteration of extracellular Na+ concentration. AB - We studied the effect of a lectin isolated from seeds of the legume Vatairea macrocarpa on net H+ efflux in Rhizobium tropici, a bacterium capable of nodulating legume Phaseolus vulgaris. V. macrocarpa lectin (VML) was observed to temporarily stimulate the specific net H(+) efflux in R. tropici. When VML was present at 32 microg ml(-1), with or without 2 microM galactose (Gal), a specific net efflux >2.4 pM H+(min)(-1) mg dry biomass(-1) was induced. There was no detectable net H+ efflux when bovine serum albumin (16 microg ml(-1)) was tested. Addition of 16 microgVMLml(-1) resulted in a 700% increase of the extracellular Na+ concentration. The soluble proteins in the supernatant containing VML extract indicate a maximum immobilization of +/-10 microgVMLml(-1), with a minimum of 36,600 dimers or 8500 larger aggregates of VML binding in each bacterium. Our data suggest that VML activates Rhizobium as a bioenergetic substrate molecule, resulting in potential alterations of the external bacterial membrane. PMID- 15336398 TI - MexZ-mediated regulation of mexXY multidrug efflux pump expression in Pseudomonas aeruginosa by binding on the mexZ-mexX intergenic DNA. AB - MexZ is a transcriptional regulator of the mexXY multidrug transporter operon, which confers aminoglycoside resistance on Pseudomonas aeruginosa. Highly purified MexZ showed direct binding with a specific site of the mexZ-mexX intergenic DNA when probed by a gel retardation assay. Both in vitro chemical cross-linking experiments and an in vivo two-hybrid expression system showed that the active form of MexZ, which is capable of binding the intergenic DNA, appeared to be a dimer. These results explain the mechanism by which MexZ represses transcription of the mexXY operon, but do not explain the substrate-induced hyperproduction of MexXY. The presence of inducer antibiotic in the gel retardation assay mixture failed to detect altered MexZ-probe DNA interaction suggesting the possible involvement of an additional regulator. PMID- 15336399 TI - Modeling the synergistic effect of high pressure and heat on inactivation kinetics of Listeria innocua: a preliminary study. AB - The survival curves of Listeria innocua CDW47 by high hydrostatic pressure were obtained at four pressure levels (138, 207, 276, 345 MPa) and four temperatures (25, 35, 45, 50 degrees C) in peptone solution. Tailing was observed in the survival curves. Elevated temperatures and pressures substantially promoted the inactivation of L. innocua. A linear and two non-linear (Weibull and log logistic) models were fitted to these data and the goodness of fit of these models were compared. Regression coefficients (R2), root mean square (RMSE), accuracy factor (Af) values and residual plots suggested that linear model, although it produced good fits for some pressure-temperature combinations, was not as appropriate as non-linear models to represent the data. The residual and correlation plots strongly suggested that among the non linear models studied the log-logistic model produced better fit to the data than the Weibull model. Such pressure-temperature inactivation models form the engineering basis for design, evaluation and optimization of high hydrostatic pressure processes as a new preservation technique. PMID- 15336400 TI - Involvement of the mpo operon in resistance to class IIa bacteriocins in Listeria monocytogenes. AB - High resistance to class IIa bacteriocins in Listeria monocytogenes has been clearly linked to lack of expression of the mptACD operon, encoding the EIIt Man mannose PTS permease. Also, intermediate resistance has been associated with membrane phospholipid modifications in the spontaneous mutants L. monocytogenes B73-V1 and B73-V2. We constructed a new mutant of L. monocytogenes that was interrupted in mpoA, and which also exhibited an intermediate resistance phenotype. The mpoABCD operon putatively encodes a PTS permease of the mannose family similar to that encoded by the mpt operon. In silico analysis indicated that mpo transcription might be dependent on sigma54. Our study demonstrated that the three intermediate resistant mutants have a slight decrease in mptACD expression, showing that the level of sensitivity is correlated to the level of mpt expression. We show a cross-regulation between mpo and mpt. In particular, the mpo mutant has a defect in mpt expression that possibly could explain its intermediate resistance phenotype. PMID- 15336401 TI - Rapid identification of pickle yeasts by fluorescent PCR and microtemperature gradient gel electrophoresis. AB - Monitoring the yeast populations within pickle soaking fluid is imperative for ensuring optimum taste, but these analyses have proven time-consuming and expensive, limiting their industrial application. Here, yeasts were identified in the soaking fluid from Japanese radish pickles using fluorescent PCR amplification of the variable D1/D2 region of the 26S rDNA, followed by analysis with microtemperature-gradient gel electrophoresis (micro-TGGE). This smaller version of the normal TGGE apparatus is capable of analyzing samples 10- to 20 fold faster without sacrificing data quality. Each primer set was labeled with a different fluorescent dye, allowing easy isolation of the various PCR products and identification of the bands corresponding to the various yeasts. The results indicate that fluorescent PCR and micro-TGGE may be a useful new method for rapid, easy monitoring of yeast flora in various food industries. This new method can be used on a daily basis to provide overviews of yeast flora during pickle production, allowing producers to quickly grasp pickle readiness at a single glance. PMID- 15336402 TI - The interaction of Pseudomonas aeruginosa PAK with human and animal respiratory tract cell lines. AB - A major virulence factor of a common human pathogen Pseudomonas aeruginosa, was investigated to determine if it dominated attachment interactions in a variety of in vitro cell culture systems. It was found that Type-IV pilus-type mechanisms, which mediated the attachment of P. aeruginosa to three human respiratory tract cell lines (A549, BEAS-2B and RPMI 2650) also mediated attachment to two respiratory tract cell lines from mouse (C57) and rat (L-2) to a similar degree. Significant differences were found in the number of P. aeruginosa associated with the human, rat and mouse cell lines. Additionally, differences were also found between A547, C57 and L-2 cells with respect to the moieties that P. aeruginosa interacted with at the level of the cell surface, suggesting that asialo-GM1 ligands were not the only structure that this bacterium could interact with in order to associate with host cells. PMID- 15336403 TI - Identification of a promoter motif regulating the major DNA damage response mechanism of Mycobacterium tuberculosis. AB - The principal response of many bacteria to DNA damage is mediated by a mechanism dependent on the LexA and RecA proteins. However, Mycobacterium tuberculosis was recently reported to regulate a majority of DNA repair genes independently of RecA and LexA, suggesting that an unknown RecA/LexA-independent mechanism controls the major DNA damage response pathway in this organism. Here we have identified a motif tTGTCRgtg-8nt-TAnnnT that defines a novel RecA/LexA independent promoter (RecA-NDp) of M. tuberculosis. Furthermore, we show that the RecA-NDp type of promoter precedes DNA repair genes in other Actinomycetales. PMID- 15336404 TI - Construction of bacterial artificial chromosome library from electrochemical microorganisms. AB - A microbial fuel cell is a device that directly converts metabolic energy into electricity, using electrochemical technology. The analysis of large genome fragments recovered directly from microbial communities represents one promising approach to characterizing uncultivated electrochemical microorganisms. To further assess the utility of this approach, we constructed large-insert (140 kb) bacterial artificial chromosome (BAC) libraries from the genomic DNA of a microbial fuel cell, which had been operated for three weeks using acetate media. We screened the expression of several ferric reductase activities in the Escherichia coli host, in order to determine the extent of heterologous expression of metal-ion-reducing enzymes in the library. Phylogenetic analysis of 16S rRNA gene sequences recovered from the BAC libraries indicates that they contain DNA from a wide diversity of microbial organisms. The constructed bacterial library proved a powerful tool for exploring metal-ion reductase activities, providing information on the electron transport pathway of electrochemical microbial (ECM) organisms. PMID- 15336405 TI - The N-terminal family 22 carbohydrate-binding module of xylanase 10B of Clostridium themocellum is not a thermostabilizing domain. AB - Xylanase Xyn10B from Clostridium thermocellum is a modular enzyme that contains two family 22 carbohydrate binding modules N- (CBM22-1) and C- (CBM22-2) terminal of the family 10 glycoside hydrolase catalytic domain (GH10). In a previous study, we showed that removal of CBM22-1 reduces the resistance to thermoinactivation of the enzyme suggesting that this module is a thermostabilizing domain. Here, we show that it is the module border on the N terminal side of GH10 that confers resistance to thermoinactivation and to proteolysis. Therefore, CBM22-1 does not function as a thermostabilizing domain and the role for this apparently non-functional CBM remains elusive. PMID- 15336406 TI - Elucidation of polysaccharide origin in Ramalina peruviana symbiosis. AB - A structural elucidation of polysaccharides extracted from the aposymbiotically cultured mycobiont of the lichen Ramalina peruviana was carried out in order to determine whether the polysaccharides found previously in the symbiotic thalli are produced by the mycobiont or photobiont or both. The mycobiont isolate was cultivated on a solid malt-yeast extract-medium and the freeze-dried colonies were defatted and the polysaccharides extracted successively with hot water and aq. 2% KOH, each at 100 degrees C. The alkaline extract was obtained in much higher yield (31.5%) and submitted to a freeze-thawing treatment, giving rise to a precipitate (PK2) of a mixture of (1-->3),(1-->4)-alpha-glucan (1.2:1 ratio, nigeran) and a (1-->3)-beta-glucan (laminaran). The mother liquor was treated with Fehling solution to give a precipitate (galactomannan). This had a (1-->6) linked alpha-d-mannopyranosyl main chain, substituted at O-4 and in small proportion at O-2,4 by beta-Galp units. All three polysaccharides have previously been found in the symbiotic thalli of R. peruviana, showing that these are produced by the fungus, without the participation of the Trebouxia photobiont. Surprisingly, isolichenan, a cold-water soluble (1-->3),(1-->4)-alpha-linked glucan (3:1 ratio) was not found in the isolated mycobiont, despite being the main polysaccharide found in the thalli. PMID- 15336407 TI - Continuous culture of Methanococcus maripaludis under defined nutrient conditions. AB - To study global regulation in the methanogenic archaeon Methanococcus maripaludis, we devised a system for steady-state growth in chemostats. New Brunswick Bioflo 110 bioreactors were equipped with controlled delivery of hydrogen, nitrogen, carbon dioxide, hydrogen sulfide, and anaerobic medium. We determined conditions and media compositions for growth with three different limiting nutrients, hydrogen, phosphate, and leucine. To investigate leucine limitation we constructed and characterized a mutant in the leuA gene for 2 isopropylmalate synthase, demonstrating for the first time the function of this gene in the Archaea. Steady state specific growth rates in these studies ranged from 0.042 to 0.24 h(-1). Plots of culture density vs. growth rate for each condition showed the behavior predicted by growth modeling. The results show that growth behavior is normal and reproducible and validate the use of the chemostat system for metabolic and global regulation studies in M. maripaludis. PMID- 15336408 TI - The cme gene of Clostridium difficile confers multidrug resistance in Enterococcus faecalis. AB - Antibiotic resistance in C. difficile by efflux has been previously suggested. The genome of C. difficile 630 was screened for sequences encoding putative proteins homologous to NorA from Staphylococcus aureus. Four ORFs homologous to efflux genes were cloned into the pAT79 shuttle vector under the control of transcription and translation signals of Gram-positive bacteria and expressed in Enterococcus faecalis JH2-2 and S. aureus RN4220. One of these sequences, designated cme conferred resistance to ethidium bromide, safranin O, and erythromycin in E. faecalis. The three other ORFs did not confer detectable resistance in both bacteria. PMID- 15336409 TI - A thermostable shikimate 5-dehydrogenase from the archaeon Archaeoglobus fulgidus. AB - Shikimate 5-dehydrogenase (SKDH; EC 1.1.1.25) catalyzes the reversible reduction of 3-dehydroshikimate to shikimate and is a key enzyme in the aromatic amino acid biosynthesis pathway. The shikimate 5-dehydrogenase gene, aroE, from Archaeoglobus fulgidus was cloned and overexpressed in Escherichia coli. The recombinant enzyme purified as a homodimer and yielded a maximum specific activity of 732 U/mg at 87 degrees C (with NADP+ as coenzyme). Apparent Km values for shikimate, NADP+, and NAD+ were estimated at 0.17+/-0.03 mM, 0.19+/-0.01 mM, and 11.4+/-0.4 mM, respectively. The half-life of the A. fulgidus SKDH is 2 h at the assay temperature (87 degrees C) and 17 days at 60 degrees C. Addition of 1 M NaCl or KCl stabilized the enzyme's half-life to approximately 70 h at 87 degrees C and approximately 50 days at 60 degrees C. This work presents the first kinetic analysis of an archaeal SKDH. PMID- 15336410 TI - Colony immunoblot assay for the detection of hemolysin BL enterotoxin producing Bacillus cereus. AB - Bacillus cereus strains involved in food poisoning cases of the diarrheal type may produce two different enterotoxin complexes. To facilitate the identification of hemolysin BL-enterotoxin complex (HBL) and/or the nonhemolytic enterotoxin (NHE) producing colonies a colony immunoblot procedure was developed, which allows a fast and easy identification of the respective colonies from blood agar plates. The enterotoxins were transferred from the blood agar medium to a nitrocellulose membrane and the immobilized toxins were probed with monoclonal antibodies. The antibodies 2A3 and 1A8 allowed the specific detection of the B component of HBL and the nheA component of NHE. The assay enabled the reliable identification of HBL expressing colonies and differentiation from NHE producing but HBL negative colonies. PMID- 15336411 TI - Diversity within Borrelia burgdorferi sensu lato genospecies in Switzerland by recA gene sequence. AB - A total of 874 Ixodes ricinus ticks were collected in Switzerland to investigate the genetic diversity of the Borrelia population. We integrated to the RT-PCR method the DNA sequence analysis of a 162-bp fragment of the recA gene. Five genospecies were detected: Borrelia afzelii, Borrelia burgdorferi s.s., Borrelia garinii, Borrelia valaisiana, and Borrelia lusitaniae. A heterogeneous distribution was observed within the B. burgdorferi s.l. genospecies. The most prevalent and diverse genospecies found in Switzerland was Borrelia afzelii, which might suggest a rapid evolution of this genospecies. PMID- 15336412 TI - Diversity of 16S rDNA sequences of Rhizobium spp. implications for species determinations. AB - Comparative analysis of 70 16S rDNA sequences representing 20 Rhizobium species (including pathogenic Agrobacterium spp.) was conducted using Maximum Likelihood to establish relationships of species using multiple sequences. There is no significant internal division of the Rhizobium clade to suggest that it represents more than one genus. Plant pathogenic (Agrobacterium) species are distributed within the genus. The analysis supported the synonymy of some species (Rhizobium gallicum and Rhizobium mongolense) and the need for comparative investigations of the tumorigenic and nodulating properties of Rhizobium tropici and Rhizobium rhizogenes. Misidentification of some sequences may conceal one or more putative novel species. Some sequences appear to be misidentified because of faulty sequencing or incomplete or inadequate analysis. PMID- 15336413 TI - Influence of ethyl acetate production and ploidy on the anti-mould activity of Pichia anomala. AB - A diploid and a haploid strain of Pichia anomala were tested for their biocontrol ability against the spoilage mould Penicillium roqueforti in glass tubes filled with grain at two water activities (aw). At aw 0.98, the two yeast strains grew and inhibited mould growth equally well and showed similar patterns of ethyl acetate production, reaching maximum values of 10-14 microg ml(-1) headspace. At aw 0.95, both growth and biocontrol performance of the haploid strain were reduced. Ethyl acetate formation was also substantially reduced, with maximum headspace concentrations of 4 microg ml(-1). We conclude that ethyl acetate is a major component of the anti-mould activity. The inhibitory effect of ethyl acetate was confirmed in a bioassay where the pure compound reduced biomass production of P. roqueforti. PMID- 15336414 TI - Salicylate decreases production of AmpC type beta-lactamases and increases susceptibility to beta-lactams in a Morganella morganii clinical isolate. AB - The effect of salicylate, a marRAB inducer, on the resistance to beta-lactams was characterized in an AmpC beta-lactamase hyperproducer Morganella morganii clinical isolate (the M1 strain). Results were compared with those of the effect of salicylate in a wild-type M. morganii strain. Salicylate induced a decreased susceptibility to nalidixic acid, norfloxacin and tetracycline and simultaneously increased the susceptibility to beta-lactams apparently due to the repression of AmpC beta-lactamase synthesis in the M1 strain. Likewise, salicylate only repressed 46 kDa outer membrane protein expression in the wild-type strain, since the clinical isolate M1 did not express it. PMID- 15336415 TI - A Myxococcus xanthus rppA-mmrA double mutant exhibits reduced uptake of amino acids and tolerance of some antimicrobials. AB - Myxococcus xanthus RppA and MmrA are homologous to methyl-accepting chemotaxis proteins (MCPs) and to multidrug transporters, respectively. We reported previously that rppA-mmrA double mutant exhibited reduced colony expansion, agglutination, and polysaccharide levels. We have demonstrated here that the rppA mmrA mutant also exhibited reduced amino acid uptake. Furthermore, the double mutant appeared to be more susceptible to some antimicrobial agents, such as streptomycin, ethidium bromide and norfloxacin, than the wild-type. These phenotypes were not shown in the rppA or mmrA single mutant. These results indicate that M. xanthus RppA and MmrA are also involved in the uptake of amino acids and efflux of some antimicrobial agents. PMID- 15336416 TI - Isolation of two aspartyl proteases from Trichoderma asperellum expressed during colonization of cucumber roots. AB - Trichoderma asperellum and cucumber seedlings were used as a model to study the modulation of Trichoderma gene expression during plant root colonization. Seedlings were grown in an aseptic hydroponics medium and inoculated with Trichoderma spore suspension. Proteins differentially secreted into the medium were isolated. Three major proteins of fungal origin were identified: two arabinofuranosidases (Abf1 and Abf2) and an aspartyl protease. Differential mRNA display was conducted on Trichoderma mycelia interacting and non-interacting, with the plant roots. Among the differentially regulated clones another aspartyl protease was identified. Sequencing of the genes revealed that the first aspartyl protease is a close homologue of PapA from T. harzianum and the other, of AP1 from Botryotinia fuckeliana. RT-PCR analysis confirms that the proteases are induced in response to plant roots attachment and are expressed in planta. papA, but not papB, is also induced in plate confrontation assays with the plant pathogen Rhizoctonia solani. These data suggest that the identified proteases play a role in Trichoderma both as a mycoparasite and as a plant opportunistic symbiont. PMID- 15336417 TI - Genetic diversity (RAPD-PCR) of lactobacilli isolated from "Almagro" eggplant fermentations from two seasons. AB - Genetic diversity of 323 strains of lactobacilli isolated from an Almagro eggplant manufacturing plant was analyzed by using random amplified polymorphic DNA (RAPD). Thirty-four distinct RAPD patterns were obtained with 95% of isolates grouped into 18 main clusters. Genetic diversity was higher in brines from season II, Lactobacillus plantarum and Lactobacillus fermentum/cellobiosus being the species with the greatest number of genotypes. A single L. fermentum/cellobiosus genotype comprised isolates from both seasons and could be considered endemic to that factory. PMID- 15336418 TI - ComX activity of Streptococcus mutans growing in biofilms. AB - In many streptococci, including Streptococcus mutans, genetic competence is regulated by a quorum sensing system mediated by a competence stimulating peptide (CSP) pheromone, encoded by the comC gene. In Streptococcus pneumoniae, a central component of this system is ComX, which acts as an alternative sigma factor to activate competence genes involved in DNA uptake and processing. The quorum sensing system responsible for genetic competence induction in S. mutans has been linked to biofilm formation and the acid tolerance response. To examine the response of comX to CSP in S. mutans, a transcriptional fusion of the comX promoter (pcomX) with lacZ was constructed to generate reporter vector pcomx::pALH122 (replicative vector) and transformed into S. mutans UA159 comC-, which is unable to produce endogenous CSP. CSP was added and pcomX::lacZ relative expression index (REI) examined, revealing a 2-fold increase in maximal beta-gal activity 5 and 10 min after CSP addition. The effect of endogenous CSP on pcomX::lacZ expression was also examined by measuring REI in cells grown as a biofilm; peak pcomX activity was observed at 3 h. To determine the temporal pattern of transformation frequency, pMA2, a Spr shuttle vector, was transformed into biofilm-grown cells, with maximal transformation frequency observed at 3 h. Confocal microscopy was performed to examine pcomX activity using a similarly constructed green fluorescent protein reporter vector, pcomX::gfp, in a 4-h biofilm, revealing active pcomX activity in high cell density areas within the biofilm population. These results demonstrated a positive correlation between pcomX activity, natural transformation and competence development in biofilms. PMID- 15336419 TI - Suppressive subtractive hybridization and differential screening identified genes differentially expressed in yeast and mycelial forms of Ophiostoma piceae. AB - Ophiostoma piceae is a sap-staining fungus that colonizes and discolours wood. It has been established that the mycelium form but not the yeast form becomes pigmented in vitro. Suppressive subtractive hybridization PCR was used to isolate transcripts specifically upregulated in either the yeast or mycelial forms of O. piceae. Subtracted cDNAs were cloned and transformed into Escherichia coli. The yeast and mycelium specific clones were then screened by differential screening to reduce the possibility of isolating false positive clones and the differential expression of the two sets of cDNAs was confirmed by reverse Northern hybridization. Numerous genes appear to be specifically expressed in either the yeast or mycelial forms. Sequence analysis identified several cDNAs similar to known genes. However, a few cDNAs showed no similarity to sequences in the public databases. PMID- 15336420 TI - Characterization of SCAR markers of Eimeria spp. of domestic fowl and construction of a public relational database (The Eimeria SCARdb). AB - This study reports the development and characterization of 151 sequence characterized amplified region (SCAR) markers for the seven Eimeria species that infect the domestic fowl. From this set, 84 markers are species-specific and 67 present partial specificity. The complete nucleotide sequence was derived for all markers, revealing the presence of micro- and minisatellite repetitive units in 22 SCARs, with up to five distinct repeat units being observed per marker. Only 15 markers showed significant hits in similarity searches against public sequence databases, thus confirming their anonymous and non-coding character. Finally, a relational database of the markers (the Eimeria SCARdb) was developed and made available on the Internet, providing a valuable resource of SCAR markers that can be useful for molecular diagnosis, and also for epizootiological, genetic variability and genome mapping studies. PMID- 15336421 TI - High numbers of Shiga toxin-producing Escherichia coli found in bovine faeces collected at slaughter in Japan. AB - The prevalence and concentration of Shiga toxin-producing Escherichia coli (STEC) in cattle faeces (n=605) at the time of slaughter was studied in Shimane Prefecture, Japan on a monthly basis between April 2000 and March 2001. Screening with stx-PCR determined a prevalence of 37.5%. After analysis of spread faeces and enriched samples on cefixime, tellurite and sorbitol-MacConkey agar using HCl treatment, 114 STEC strains were singly or concomitantly isolated from 97 cattle (15.9%). Of the 605 cattle, 31 (5.1%) harbored O26:H11, O111:H-, O121:H19 or O157:H7, which had the stx1 and/or stx2 and eae and hlyA genes, and 7 (23%) of these 31 cattle were high level carriers that contained these typical STEC at concentrations of 10(5)-10(8) CFU/g of faeces. The predominant serotype was O26:H11 (20 strains) and the second most frequent was O157:H7 (9 strains). Of the 605 cattle, 68 (11.2%) harboured 36 other serotypes and 6 (5.9%) of the 67 cattle were high level carriers. As a comparison between the prevalence of STEC and the faecal pH, it was demonstrated that STEC can be isolated from cattle with a wide range of faecal pH values. The presence of a high-carriage animal at the abattoir increases the potential risk of meat contamination during the slaughtering process, regardless of faecal pH. PMID- 15336422 TI - Characterization of the Azotobacter vinelandii algC gene involved in alginate and lipopolysaccharide production. AB - Azotobacter vinelandii is a soil gamma-proteobacteria that fixes nitrogen and forms desiccation-resistant cysts. The exopolysaccharide alginate is an integral part of the layers surrounding the cysts. Here, we reported the cloning of A. vinelandii algC, encoding the enzyme catalyzing the second step of alginate pathway. We showed that AlgC is involved not only in alginate production, but also in lipopolysaccharide (LPS) synthesis and that it seems to have both phosphomannomutase and phosphoglucomutase activities. The transcriptional analysis of the A. vinelandii algC gene showed that it contained two start sites, one of which was dependent on the alternative sigma factor AlgU/AlgT. This finding explains why alginate biosynthesis is dependent on AlgU activity, since all other alginate biosynthetic genes have been characterized previously and algC is the only alginate structural gene that is directly transcribed by this sigma factor. PMID- 15336423 TI - A mannose binding protein is involved in the adherence of Acanthamoeba species to inert surfaces. AB - Some carbohydrates are known to decrease the attachment of Acanthamoeba sp. to biological surfaces. By a method based on the reduction of a tetrazolium salt (XTT) by the mitochondrial dehydrogenases of the parasites, d-mannose and alpha-d mannopyranoside have been shown to reduce Acanthamoeba attachment to inert surfaces, indicating that the mannose binding protein of Acanthamoeba trophozoites is involved in adherence to inert surfaces. The reduction in attachment is dose dependant and is not linked with a potential toxicity of the carbohydrates. All the species of Acanthamoeba tested were concerned by this mannose binding protein, but the adhesion of A. culbertsoni was also reduced by the presence of glucose. PMID- 15336424 TI - Characterisation of a new Leishmania META gene and genomic analysis of the META cluster. AB - The META1 gene of Leishmania is upregulated in metacyclic promastigotes and encodes a 12 kDa virulence-related protein, conserved in all Leishmania species analysed. In this study, the genomic region adjacent to the Leishmania amazonensis META1 gene was characterised and compared to the Leishmania major META1 locus as well as to syntenic loci identified in Trypanosoma brucei and Trypanosoma cruzi. Three new genes expressed with increased abundance of steady state mRNA in L. amazonensis promastigotes were identified, two of which are upregulated in stationary phase promastigotes, sharing the pattern of expression previously described for the META1 mRNA. One of these new genes, named META2, encodes a polypeptide of 444 amino acid residues with a repetitive structure showing three repeats of the META domain (defined as a small domain family found in the Leishmania META1 protein and in bacterial proteins hypothetically secreted and/or implicated in motility) and a carboxyl-terminal region similar to several putative calpain-like proteins of Trypanosoma and Leishmania. PMID- 15336425 TI - Variation in antimicrobial activity of lactoferricin-derived peptides explained by structure modelling. AB - Antimicrobial peptides bovine lactoferricin (LfcinB) and human lactoferricin (LfcinH) are produced from the respective lactoferrin, but are more active than their precursors. Despite sequence homology, the bovine peptide and its derivatives are more active than their human homologs. Such differences between not only the peptides and their precursor but also between the bovine and the human peptides could relate to structural differences. Upon sequence alignment of both peptides with their parental proteins, the structural differences observed between the bovine lactoferrin (BLf) and LfcinB were also found between the human lactoferrin (HLf) and the LfcinH. The helical structures in HLf are replaced by beta-strands separated by a strong turn in LfcinH suggesting an antiparallel beta sheet structure similar to LfcinB. MIC assays with HLP-2 and BLP-2, 11-residue peptides derived from the active core of both Lfcins, against Escherichia coli, showed that the bovine derivative, BLP-2, is more active than its human homolog HLP-2. Both 3D models for HLP-2 and BLP-2 showed that the beta-strand is centred between the aromatic residues giving both side chains the same orientations. The displacement towards the N-terminus observed for the beta-strand in HLP-2, compared with its central location in BLP-2, could be less favourable to membrane interaction and therefore responsible for the decrease in activity. Such a model suggests for LfcinH a mechanism similar to the one observed for LfcinB, where the absence of long-range interaction, present in lactoferrin, destabilises the first alpha helix, as observed in solution and, upon interaction with the membrane, could result in the formation of a beta-strand, as observed in the presence of LPS. The location of the beta-strand in relation to the positive charges, seems to define the efficiency of the activity of the peptide and may explain the difference in activity obtained between HLP-2 and BLP-2. PMID- 15336426 TI - Identification of nucleotides critical for activity of the sigmaE-dependent rpoEp3 promoter in Salmonella enterica serovar Typhimurium. AB - We previously described a two-plasmid system for the identification of promoters recognized by Salmonella enterica serovar Typhimurium (S. Typhimurium) sigmaE. The S. Typhimurium sigmaE-dependent rpoEp3 promoter was active in the E. coli two plasmid system only after arabinose-induced expression of S. Typhimurium rpoE. In the present study, we have exploited this two-plasmid system for the identification of nucleotides critical for activity of the rpoEp3 promoter. A library of randomly mutated DNA fragments containing the rpoEp3 promoter was cloned upstream of a lacZalpha reporter gene and screened for activity in the presence of S. Typhimurium sigmaE. The clones exhibiting reduced LacZ activity were sequenced to identify the mutations. The activity of the mutated rpoEp3 promoters were studied further using a luciferase-based promoter-probe plasmid. All of the important nucleotides of the rpoEp3 promoter (in capital) were located in the -35 (ggAActt) and -10 (TctaA) regions. The critical nucleotides were also the most conserved in known sigmaE-dependent promoters. The study also revealed the importance of the 16-bp spacing between -10 and -35 region, as reducing the spacing to 15-bp greatly reduced activity of the promoter. This method should be generally applicable for the identification of important nucleotides in the cognate promoters of other sigma factors. PMID- 15336427 TI - Pichia anomala and Kluyveromyces wickerhamii killer toxins as new tools against Dekkera/Brettanomyces spoilage yeasts. AB - Two yeast killer toxins active on spoilage yeasts belonging to the genus Dekkera/Brettanomyces are here described for the first time. The two toxins produced by Pichia anomala (DBVPG 3003) and Kluyveromyces wickerhamii (DBVPG 6077), and named Pikt and Kwkt, respectively, differ for molecular weight and biochemical properties. Interestingly, the fungicidal effect exerted by Pikt and Kwkt against Dekkera bruxellensis is stable for at least 10 days in wine. Thus, a potential application for the two toxins as antimicrobial agents active on Dekkera/Brettanomyces during wine ageing and storage can be hypothesised. PMID- 15336428 TI - Low concentrations of the non-ionic detergent Nonidet P-40 interfere with sterol biogenesis and viability of the yeast Saccharomyces cerevisiae. AB - Mild non-ionic detergents are used for solubilization of hydrophobic substrates in yeast growth media at concentrations 0.1-1%. Our data show that low concentrations of Nonidet P-40 may significantly affect lipid biogenesis in the yeast Saccharomyces cerevisiae. The uptake and esterification of external [4-14C] cholesterol is strongly reduced in hem1 mutants treated with low concentrations of Nonidet P-40. Significant inhibitory effect of NP-40 on sterol uptake and esterification was evident both in non-growing and growing cells supplemented with external cholesterol. Increased levels of sterol precursors (squalene, lanosterol) in hem1 cells grown in complex medium with cholesterol indicated general interference of NP-40 with sterol biosynthesis. NP-40 in the growth medium affected also cell viability estimated as the colony forming ability. More attention should be therefore paid to possible effects of mild detergents at low concentrations generally considered to be harmless, especially in cells with disturbed lipid biogenesis. PMID- 15336429 TI - A rubrerythrin-like oxidative stress protein of Clostridium acetobutylicum is encoded by a duplicated gene and identical to the heat shock protein Hsp21. AB - Comparison of the N-terminus of the heat shock protein Hsp21 of Clostridium acetobutylicum with proteins predicted to be encoded by the genome of this bacterium revealed that this stress protein is encoded by two almost identical open reading frames CAC3597 and CAC3598. These genes encode a rubrerythrin-like protein with the rubredoxin-like FeS4 domain at the N-terminus and the ferritin like diiron domain (rubrerythrin domain) at the C-terminus. Thus, the order of the two putative functional domains is reversed compared to "normal" rubrerythrins. This protein is proposed to be involved in the oxidative stress response of strict anaerobic bacteria. Northern blot analysis indicated that hsp21 is induced by heat and oxidative stress (air, H2O2). Hsp21 of C. acetobutylicum can be considered as a "reverse" rubrerythrin and a role of this stress protein, which is conserved among clostridia and other strict anaerobic bacteria, in the heat and oxidative stress response is proposed. PMID- 15336430 TI - Characterisation of complex formation between members of the Mycobacterium tuberculosis complex CFP-10/ESAT-6 protein family: towards an understanding of the rules governing complex formation and thereby functional flexibility. AB - We have previously shown that the secreted M. tuberculosis complex proteins CFP 10 and ESAT-6 form a tight, 1:1 complex, which may represent their functional form. In the work reported here a combination of yeast two-hybrid and biochemical analysis has been used to characterise complex formation between two other pairs of CFP-10/ESAT-6 family proteins (Rv0287/Rv0288 and Rv3019c/Rv3020c) and to determine whether complexes can be formed between non-genome paired members of the family. The results clearly demonstrate that Rv0287/Rv0288 and Rv3019c/3020c form tight complexes, as initially observed for CFP-10/ESAT-6. The closely related Rv0287/Rv0288 and Rv3019c/Rv3020c proteins are also able to form non genome paired complexes (Rv0287/Rv3019c and Rv0288/Rv3020c), but are not capable of binding to the more distantly related CFP-10/ESAT-6 proteins. PMID- 15336431 TI - Comparison of three microscopic techniques for diagnosis of Cyclospora cayetanensis. AB - Cyclospora cayetanensis oocysts in the feces of humans from Kathmandu, Nepal were identified on the basis of their size and other morphological characteristics. We compared the detection of C. cayetanensis oocysts in the feces using three microscopic techniques such as formalin-ether sedimentation, sucrose centrifugal floatation, and direct smear. Standard procedures were used for the formalin ether sedimentation and the sucrose centrifugal floatation techniques using 0.5 g of feces, however, the direct smear technique was performed using 10 microl of fecal suspension (0.005 g of feces) and observed under the fluorescent microscope. Of the 403 samples examined, 21 samples were positive for oocysts by all three techniques. Therefore, in these 21 samples, the number of oocysts recovered by the three techniques were compared. The highest number of oocyst was obtained by the sucrose centrifugal floatation technique. In contrast, the formalin-ether sedimentation technique was found to be the least reliable concentration technique for the detection of Cyclospora in human feces. Surprisingly, the direct smear technique was found to be an effective and rapid technique for diagnosis of C. cayetanensis making it a technique of choice for routine epidemiological investigation of the prevalence of this infection in human populations. PMID- 15336432 TI - Role of an acrR mutation in multidrug resistance of in vitro-selected fluoroquinolone-resistant mutants of Salmonella enterica serovar Typhimurium. AB - Quinolone resistance in Salmonella spp. is usually attributed to both active efflux and mutations leading to modification of the target enzymes DNA gyrase and topoisomerase IV. Here, we investigated the presence of mutations in the efflux regulatory genes of fluoroquinolone- and multidrug-resistant mutants of Salmonella enterica serovar Typhimurium (S. Typhimurium) selected in vitro with enrofloxacin that both carried a mutation in the target gene gyrA and overproduced the AcrAB efflux pump. No mutations were detected in the global regulatory loci marRAB and soxRS for the four strains studied. A mutation in acrR, the local repressor of acrAB, was found for two ciprofloxacin-resistant selected-mutants, leading to duplication of amino acids Ile75 and Glu76. Complementation experiments with wild-type acrR showed that the mutation identified in acrR partially contributed to the increase in resistance levels to several unrelated antibiotics. The acrR mutation also contributed to acrAB overexpression as shown by RT-PCR. Thus, this study underlines the role of an acrR mutation, in addition to the mutation in gyrA, in the fluoroquinolone and multidrug resistance phenotype of S. Typhimurium mutants, through overexpression of acrAB. PMID- 15336433 TI - [Lipidomies is energing: 1st and 2nd lipidomics meeting]. PMID- 15336434 TI - [Macrophage activation syndrome, hemophagocytic syndrome]. AB - Macrophage activation syndrome MAS describes the clinical, biological and histological symptoms related to a probably T lymphocytes/NK cell driven stimulation of macrophages with the consequence of a hemophagocytosis involving numerous organs, preferentially bone marrow, explaining the other term of "hemophagocytic syndrome". Clinical symptoms include cytopenia, multiple organ dysfunction, fever unresponsive to antibiotics, fatigue and rash. Infections (bacteria, virus or parasites), lymphoproliferative disorders, cancers, systemic diseases are the most prevalent triggers or etiologies of M.A.S. Evidence of haemaphagocytosis is obtained in the majority of cases with bone marrow specimens. In some cases haemophagocytosis can spare the bone marrow with involvement confined to other tissues such as liver and spleen. Very high levels of ferritine seem to correlate well with the presence of haemophagocytosis and is a possible marker for an early diagnosis. Early treatment initiation is mandatory. Corticosteroids, cytostatic drugs such as etoposide, cyclosporine A, plasmapherese, intravenous immunoglobulins and anti TNFalpha are proposed but no randomized trials were published. PMID- 15336435 TI - [Pathophysiological role of endotoxins, a common denominator to various diseases]. AB - A growing number of investigations point to endotoxin or lipopolysaccharide as a central player in many pathophysiological states and diseases. Endotoxins are one of the most toxic biological contaminants continuously shed by both dead and live Gram negative bacteria. Endotoxins induce the primitive form of defense called innate immunity. Endotoxins have been related to inflammatory reactions observed in patients suffering from respiratory distress syndrome, multiorgan failure and septic shock, hepatic diseases, or in subjects affected by graft versus host disease after allogeneic transplantation. As our understanding of the molecular mechanisms underlying pathologies progresses, more diseases involving endotoxins emerge. Although these illnesses are multifactorial, the objective of this article is to review some of the common and distinct processes involving endotoxins in various disease states. PMID- 15336437 TI - Mechanisms of hepatitis B virus graft reinfection and graft damage after liver transplantation. PMID- 15336438 TI - The effects of oxidative stress on the liver sieve. AB - BACKGROUND/AIMS: Oxidative stress is implicated in the pathogenesis of age related and disease-associated changes in the hepatic sinusoid. We studied the effects of oxidative stress on the morphology of the liver, focusing specifically on the hepatic sinusoidal endothelium (the 'liver sieve'). METHODS: The effects of tert-butyl hydroperoxide on the intact liver and isolated sinusoidal endothelial cells were assessed by electron microscopy, immunohistochemistry and biochemical analysis. RESULTS: Immunohistochemistry revealed a dose-dependent increase in peri-sinusoidal 3-nitrotyrosine staining, particularly in the regions adjacent to the portal triads. Electron microscopy showed dose-dependent formation of large intracellular gaps in the sinusoidal endothelium with reduction in the diameter of the remaining endothelial fenestrations. Activated Kupffer cells extending processes through the fenestrations to contact hepatocytes were noted. Biochemical analysis of total liver tissue showed no significant changes in malondialdehyde content but a decrease in the ratio of GSH to GSSG. tert-Butyl hydroperoxide administered directly onto isolated liver sinusoidal endothelial cells was associated with similar gap formation, indicating a direct effect on the endothelial cells by tert-butyl hydroperoxide. CONCLUSIONS: Oxidative stress selectively damages hepatic sinusoidal endothelial cells. This has implications for those processes associated with changes in the sinusoidal endothelium such as ageing, cirrhosis and exposure to hepatotoxins. PMID- 15336439 TI - High-fat enteral nutrition reduces endotoxin, tumor necrosis factor-alpha and gut permeability in bile duct-ligated rats subjected to hemorrhagic shock. AB - BACKGROUND/AIMS: Cholestatic patients are prone to septic complications after major surgery due to an increased susceptibility to endotoxin and hypotension. High-fat enteral nutrition reduces endotoxin after hemorrhagic shock. However, it is unknown whether this nutritional intervention is protective in biliary obstruction. We investigated the effect of high-fat enteral nutrition on endotoxin, tumor necrosis factor-alpha (TNF-alpha) and intestinal permeability in cholestatic rats subjected to hemorrhagic shock. METHODS: Bile duct-ligated (BDL) rats were fasted or fed with low-fat or high-fat enteral nutrition before hemorrhagic shock. Blood and tissue samples were taken after 90 min. RESULTS: Plasma endotoxin decreased after hemorrhagic shock in BDL-rats fed with high-fat nutrition compared to fasted (P<0.01) and low-fat treated rats (P<0.05). Additionally, circulating TNF-alpha was reduced in BDL-rats pretreated with high fat nutrition compared to fasted rats (P<0.01). The increased intestinal permeability to macromolecules was reduced by high-fat enteral nutrition, whereas bacterial translocation did not significantly change. Simultaneously, tight junction distribution in ileum and colon was disrupted in non-treated BDL-rats but remained unchanged in high-fat pretreated BDL-rats. CONCLUSIONS: High-fat enteral nutrition protects against endotoxin-mediated complications independently of intraluminal bile. These results provide a potential new strategy to prevent endotoxin-mediated complications in cholestatic patients undergoing major surgery. PMID- 15336440 TI - Effect of intravenous albumin on systemic and hepatic hemodynamics and vasoactive neurohormonal systems in patients with cirrhosis and spontaneous bacterial peritonitis. AB - BACKGROUND/AIMS: Albumin administration prevents renal failure and improves survival in spontaneous bacterial peritonitis. This study characterizes the mechanisms of action of albumin in this condition. METHODS: Systemic and splanchnic hemodynamics, plasma renin activity and plasma concentration of interleukin-6, serum concentration of nitric oxide and ascitic fluid levels of nitric oxide and interleukin-6 were assessed at diagnosis and resolution of infection in 12 patients with spontaneous bacterial peritonitis treated with ceftriaxone plus albumin. At infection resolution there was a significant improvement in circulatory function, as indicated by a significant increase in mean arterial pressure (+8%, P=0.02), a fall in heart rate (-10%, P=0.01), a suppression of plasma renin activity (-67%, P=0.002) and a decrease in creatinine levels. These changes were related to both an increase in cardiac work (stroke work index: +18%, P=0.005) and in peripheral vascular resistance (+14%, P=0.05). The improvement in cardiac function was due to an increase in filling. No significant changes were observed in portal pressure or hepatic blood flow. CONCLUSIONS: These results indicate that the beneficial effects of albumin administration on systemic hemodynamics and renal function in spontaneous bacterial peritonitis are related to both an improvement in cardiac function and a decrease in the degree of arterial vasodilation. PMID- 15336441 TI - A novel aminosterol reverses diabetes and fatty liver disease in obese mice. AB - BACKGROUND/AIMS: Non-alcoholic fatty liver disease (NAFLD) is common in obesity. However, weight reduction alone does not prevent the development or progression of NAFLD. Since NAFLD is associated with insulin resistance and diabetes, we hypothesized that improvement of these factors would reverse obesity-related NAFLD. METHODS: We examined the effects of an aminosterol, 1436, on glucose, lipids and liver metabolism in Lep(ob/ob) mice, a model of obesity, severe insulin resistance, diabetes, hyperlipidemia and hepatic steatosis. RESULTS: 1436 decreased body weight, specifically fat content, by inhibiting food intake and increasing energy expenditure. In contrast to weight loss from food restriction, this aminosterol specifically lowered circulating lipids, reversed hepatic steatosis and normalized alanine aminotransferase level. 1436 decreased glucose, increased adiponectin and enhanced insulin action in liver. These changes culminated in inhibition of hepatic triglyceride synthesis and increased fatty acid oxidation. Gene expression studies confirmed a reduction in lipogenic enzymes in liver, and elevation of enzymes involved in lipid catabolism. CONCLUSIONS: These results demonstrate that 1436 is an effective treatment for insulin resistance and hepatic steatosis in Lep(ob/ob) mice, by decreasing hepatic lipid synthesis and stimulating lipolysis. In contrast, weight loss from food restriction has no substantial effect on insulin resistance, lipids and hepatic steatosis. PMID- 15336442 TI - A gene-expression signature can quantify the degree of hepatic fibrosis in the rat. AB - BACKGROUNDS/AIMS: A more accurate and objective quantification of hepatic fibrosis would provide clinically useful information for the monitoring of chronic liver disease progression and therapy recommendation. METHODS: Using a cDNA microarray of 14,814 clones, we analyzed the gene-expression profiles of fibrotic livers in a rat model. RESULTS: We identified 750 up- and 345 down regulated genes by combining a signal-to-noise score and a random permutation test (P<0.01). The functions of these genes provided insight into the underlying molecular mechanisms of both structural remodeling and functional deficits in cirrhosis. To quantify the extent of liver fibrosis, we have generated for the first time a 'genetic fibrosis index' based on gene-expression profiling of 95 genes by combining a Pearson correlation coefficient and a 'leave-one-out' cross validation procedure. This technique based on a supervised learning analysis correctly quantified the various degrees of fibrosis in both 20 training samples (R(2)=0.829, P<0.001) and 6 test samples (R(2)=0.822, P<0.05). CONCLUSIONS: Our method will assist researchers in identifying rational targets for intervention and might help clinicians to objectively monitor the severity of liver fibrosis. PMID- 15336443 TI - Molecular mechanisms regulating the antifibrogenic protein heme-oxygenase-1 in human hepatic myofibroblasts. AB - BACKGROUND/AIMS: Hepatic myofibroblasts are central in liver fibrogenesis associated with chronic liver diseases. We previously showed that heme-oxygenase 1 (HO-1) displays antifibrogenic properties in human hepatic myofibroblasts. Here, we further investigated the mechanisms regulating HO-1 expression. METHODS: Expression of HO-1 was assayed in cultured human hepatic myofibroblasts by Northern and Western blot. Functional studies were also performed in cultured human hepatic myofibroblasts. RESULTS: 15-Deoxy-Delta(12,14)-prostaglandin J2 (15 d-PGJ2) elicited inhibition of proliferation and of alpha1(I) collagen mRNA expression. These effects were reproduced by the glutathione depletor diethyl maleate and blunted by the glutathione precursor N-acetyl cysteine, indicating the involvement of oxidative stress. Two consecutive events mediated inhibition of proliferation and of alpha1(I) collagen mRNA expression by 15-d-PGJ2: (i) mild oxidative stress characterized by a transient GSH decrease and (ii) activation of p38 MAPK, resulting in increased HO-1 mRNA stability. CONCLUSIONS: Our results provide new insights into the regulatory mechanisms governing HO-1 expression in human hepatic myofibroblasts and identify mild oxidative stress and p38 MAPK as two consecutive early signals promoting HO-1 induction that are crucial for its antifibrogenic properties, namely inhibition of growth and extracellular matrix gene expression. PMID- 15336444 TI - Protein kinase A dependent signalling mediates anti-apoptotic effects of the atrial natriuretic peptide in ischemic livers. AB - BACKGROUND/AIMS: Preconditioning of livers with atrial natriuretic peptide (ANP) attenuates ischemia-reperfusion injury (IRI) via the particulate guanylate cyclase. Recently, we have shown that ANP affects the p38 MAPK signalling cascade in the liver. Thus, aim of the present study was to elucidate the role of cGMP- and p38 MAPK-dependent signalling pathways in ANP-mediated anti-apoptotic effects. METHODS: Rat livers were perfused with KH-buffer with or without ANP, 8 Br-cGMP (+/-kinase inhibitors) and kept in UW solution (4 degrees C, 24h). Caspase-3-like activity was measured by a fluorometric assay. Expression of cGMP dependent protein kinases (PKG) in liver tissue was determined by RT-PCR, BAD phosphorylation by Western blot, and cAMP-dependent protein kinase (protein kinase A, PKA) activity by in vitro phosphorylation. RESULTS: Compared to control organs, ANP-preconditioning reduced post-ischemic caspase-3-like activity. Neither perfusion with a p38 MAPK inhibitor nor with a PKG inhibitor abolished the ANP-mediated anti-apoptotic action. The two PKG isoforms were demonstrated not to be expressed in the liver. In contrast, liver perfusion with a selective PKA inhibitor abrogated the anti-apoptotic effect of ANP. Phosphorylation of pro apoptotic BAD by ANP-activated PKA might inhibit liver cell apoptosis. CONCLUSIONS: ANP mediates its anti-apoptotic action during ischemic injury via a crosstalk with the PKA pathway. PMID- 15336445 TI - Usefulness of contrast-enhanced perfusional sonography in the assessment of hepatocellular carcinoma hypervascular at spiral computed tomography. AB - BACKGROUND/AIMS: Diagnosis of hepatocellular carcinoma (HCC) relies strongly on the detection of hypervascularity in the arterial phase and, in this setting, spiral computed tomography (CT) is the most widely used method. This prospective study aimed to investigate the usefulness of low mechanical index harmonic ultrasound (US), using a second generation contrast-enhanced technique, in the assessment of vascular pattern of HCC shown to be hypervascular at spiral CT. METHODS: A total of 79 cirrhotic patients with 103 nodules (mean+/-SD 28+/-13 mm) with arterial hypervascularity at spiral CT were studied. US examination was performed by perfusional sonography, using a new dedicated technology (CnTI Esaote trade mark ), operating at low mechanical index, after injection of a second generation contrast agent (SonoVue trade mark -Bracco), allowing detection of tumoral flow during arterial phase. RESULTS: Selective arterial enhancement on perfusional sonography was observed in 94 /103 nodules (91.3%), with a sensitivity of 66.6, 87.5, 91.7, and 97.3% in nodules 123 cm respectively. CONCLUSIONS: Perfusional sonography shows good diagnostic agreement with spiral CT in hypervascular HCC and may be proposed for the immediate vascular characterization of nodules detected at US and used as second imaging technique to confirm hypervascularity in cirrhotic nodules. PMID- 15336446 TI - Transarterial chemo-lipiodolization can reactivate hepatitis B virus replication in patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: Reactivation of hepatitis B virus (HBV) replication is a well known complication in cancer patients receiving chemotherapy. The aims of this study were to determine the incidence of HBV reactivation in hepatocellular carcinoma (HCC) patients undergoing transarterial chemo-lipiodolization, and to clarify factors contributing to HBV reactivation. METHODS: From April 2001 to September 2002, 146 HBsAg positive patients newly diagnosed as HCC were enrolled in the study. Among these, 83 patients underwent transarterial chemo lipiodolization using epirubicin and/or cisplatin, and 63 received other treatments. RESULTS: In total, HBV reactivation occurred in 30 (20.5%) patients (28 with chemo-lipiodolization and 2 with other treatments), and of the 30 patients, 19 (13.0%) (18 with chemo-lipiodolization and 1 with other treatments) developed hepatitis. Chemo-lipiodolization was significantly correlated with a higher incidence of hepatitis attributed to HBV reactivation than other treatments (21.7% vs. 1.6%, P<0.001), irrespective of HBeAg or HBV DNA. Among 83 patients undergoing chemo-lipiodolization, HBV reactivation occurred in 28 (33.7%) patients, and HBeAg seropositivity was the only independent predictor of HBV reactivation (P=0.013). Three (10.7%) of them died of hepatic decompensation resulting from HBV reactivation. CONCLUSIONS: Transarterial chemo-lipiodolization can reactivate HBV, and HBeAg-positive HCC patients receiving chemo lipiodolization should be closely monitored for HBV reactivation. PMID- 15336447 TI - Identification of genes up-regulated by histone deacetylase inhibition with cDNA microarray and exploration of epigenetic alterations on hepatoma cells. AB - BACKGROUND/AIMS: Epigenetics is the key factor in the regulation of gene expression. We conducted cDNA microarray analysis to screen for genes induced by histone deacetylase (HDAC) inhibition and examined epigenetic alterations. METHODS: Microarray analysis was performed in six hepatoma cell lines and primary hepatocytes treated with trichostatin A (TSA). mRNA expression of several genes was examined by reverse transcription-polymerase chain reaction in TSA-treated cells and hepatoma samples. Acetylated histones and methylation status in 5'CpG islands was assessed by chromatin immunoprecipitation (ChIP) assay and bisulfite genomic sequencing, respectively. RESULTS: Fifty-seven genes showed greater than 2-fold change after TSA treatment in multiple cell lines. Among them, four genes including p21(WAF1) exhibited substantial induction (greater than 5-fold changes). Decreased mRNA levels of these genes in hepatoma tissues were observed in more than half of patients. ChIP assay, in general, demonstrated a good correlation between mRNA expression and histone acetylation, but only a limited correlation with the methylated DNA in the promoter region. CONCLUSIONS: We identified 57 up-regulated genes by TSA treatment in hepatoma cells and some of them appeared to be cancer-related genes in hepatomas. The alterations in acetylated histones are likely closely associated with gene expression. PMID- 15336448 TI - Donor age influences 10-year liver graft histology independently of hepatitis C virus infection. AB - BACKGROUND/AIMS: Factors influencing the long-term histological outcome of liver graft are not known. We conducted a prospective study based on a 10-year liver biopsy in order to identify the main factors influencing long-term graft histology. METHODS: 270 of 423 patients who still had their first functional graft 10 years after liver transplantation accepted to undergo routine liver biopsy. All slides were blindly reviewed by two pathologists. RESULTS: Main histological findings were fibrosis in 143 patients (54%) and ductopenia in 76 patients (29%). Ductopenia was independently related to higher donor age (32+/-12 vs 28+/-13 years; P<0.02). Severity of fibrosis was influenced by hepatitis C virus (HCV) infection (P<0.001), hepatitis B virus (HBV) recurrence (P=0.001) and higher donor age (P=0.03). Eighty biopsies (30%) showed minimal-change lesions which were associated with the absence of HCV infection (24/80 vs 99/185; P<0.001) or of HBV infection (1/80 vs 15/185; P=0.03) and lower donor age (25+/ 11 vs 31+/-13 years; P<0.001). CONCLUSIONS: Post-transplant infection by HCV or HBV are main factors influencing the histological course of liver graft. Donor age was also a strong factor in HCV infected patients as well as in HCV-negative patients. This variable should be taken into account, particularly for candidate recipients with long life expectancy. PMID- 15336449 TI - Comparison of clinical outcome between patients continuing and discontinuing lamivudine therapy after biochemical breakthrough of YMDD mutants. AB - BACKGROUND/AIMS: The aim of this study was to compare the clinical outcome between patients continuing and discontinuing lamivudine therapy after the biochemical breakthrough of hepatitis B virus tyrosine-methionine-aspartate aspartate (YMDD) mutant. METHODS: YMDD mutants were detected in 51 chronic hepatitis B patients who experienced a flare-up of alanine aminotransferase (ALT) during lamivudine treatment. Twenty-seven of them discontinued lamivudine therapy (group A), and 24 continued therapy (group B) after biochemical breakthrough. The follow-up period was 12 months in both the groups. RESULTS: There was no significant difference between groups A and B in the incidence and severity of ALT peaks and hepatic decompensation within the first 3 months after biochemical breakthrough. After the fourth month of biochemical breakthrough, however, group A experienced acute exacerbation more frequently [20/26 (77%) vs. 7/23 (30%); P=0.002] and higher ALT peaks than group B. The same result was found when the patients were divided into naive and retreated or cirrhotic and non-cirrhotic groups. Hepatic decompensation at the onset of biochemical breakthrough was associated with higher mortality (OR=70, 95% CI=6.06-807.75). CONCLUSIONS: Patients who discontinued lamivudine therapy increased the frequency of flare-ups and higher ALT peaks than those who continued therapy after 4 months post breakthrough. PMID- 15336450 TI - Evaluation of amantadine in chronic hepatitis C: a meta-analysis. AB - BACKGROUND/AIMS: The benefit of amantadine combination therapy, either with interferon (IFN) alone (double therapy) or with ribavirin and IFN (triple therapy) is unknown. METHODS: We analyzed the effect of amantadine on the end-of treatment virological response and the sustained response using meta-analysis of 31 randomized controlled trials. RESULTS: Overall analysis revealed a significant effect of amantadine. Triple therapy was the best regimen for improving the sustained response (mean difference: 8.4%, 95% CI: 2.4-13.8%, P=0.002). In subgroup analysis, amantadine did not have a significant effect upon naive patients or relapsers. In non-responders, combination therapy with amantadine was associated with a significant effect on the sustained response (mean difference: 8.3%, 95% CI: 1.9-14.6%, P=0.01). In sensitivity analysis, double therapy did not improve virological responses. Conversely, triple therapy tended to improve the end-of-treatment virological response and was associated with a significant effect upon the sustained response (mean difference: 12.7%, 95% CI: 3.8-21.6%, P=0.005). CONCLUSIONS: Combination therapy with amantadine is of no effect upon naive patients or relapsers. In non-responders, triple therapy with amantadine improved the sustained response. New randomized controlled trials are required to confirm this meta-analysis. PMID- 15336451 TI - Peginterferon alfa-2b plus ribavirin for naive patients with genotype 1 chronic hepatitis C: a randomized controlled trial. AB - BACKGROUND/AIMS: We assessed the effectiveness and safety of an induction dose of peginterferon alfa-2b (PEG-IFN) plus ribavirin for initial treatment of patients with genotype 1 chronic HCV infection in a randomized, controlled, multicenter trial. METHODS: Three hundred and eleven naive patients infected with genotype 1 and chronic hepatitis were randomly assigned to 48-week treatment with PEG-IFN once weekly (80-100 micrograms depending on body weight for 8 weeks, followed by 50 micrograms for the next 40 weeks), or standard interferon alfa-2b (IFN) 6 million units on alternate days, both in combination with ribavirin (1000-1200 mg/day). RESULTS: PEG-IFN plus ribavirin significantly increased sustained virological response (SVR) compared with IFN plus ribavirin (41.1 vs. 29.3% respectively, P=0.030). Less patients discontinued PEG-IFN than IFN (19 vs. 31%, P=0.010). By logistic regression, SVR in the PEG-IFN group was independently associated with age <50 years, and mild fibrosis at liver biopsy. CONCLUSIONS: Combination therapy with an induction dose of PEG-IFN was a more effective and better tolerated treatment for naive patients with genotype 1 than combination therapy with high dose standard IFN. In patients aged less than 50 years with mild fibrosis this schedule achieves a very high rate of SVR. PMID- 15336452 TI - Out of control: hepatitis B reactivation by transarterial treatment of hepatocellular carcinoma. PMID- 15336453 TI - Older livers--should they be used for transplantation? PMID- 15336454 TI - Induction therapy in chronic hepatitis C: deja-vu with pegylated interferons? PMID- 15336455 TI - Telomeres and telomerase: new targets for the treatment of liver cirrhosis and hepatocellular carcinoma. PMID- 15336456 TI - Liver inflammatory pseudotumor due to an intrahepatic wooden toothpick. PMID- 15336457 TI - Liver nodules ressembling focal nodular hyperplasia after portal vein thrombosis. PMID- 15336458 TI - Prompt relapse of viremia after lamivudine discontinuation in e-minus chronic hepatitis B patients completely responders during 5 years of therapy. PMID- 15336459 TI - Harmonization of liver enzyme results: calibration for aminotransferases and gamma glutamyltransferase. PMID- 15336460 TI - Coarse vs. fine needle aspiration biopsy. PMID- 15336463 TI - Safety profile of oral and intravenous moxifloxacin: cumulative data from clinical trials and postmarketing studies. AB - BACKGROUND: The established safety profile of the fluoroquinolones has been disrupted in the past decade by the detection of low-frequency but potentially serious adverse events that have led to the license suspension, voluntary withdrawal, or restricted use of specific members of the class. Moxifloxacin is a broad-spectrum, advanced-generation fluoroquinolone that has potent activity against respiratory tract infections in adults in both oral and IV formulations. OBJECTIVE: The goal of this article was to provide an overview of the cumulative safety data on both oral and IV moxifloxacin, including data from the most recent clinical trials and postmarketing studies. METHODS: Data from clinical trials of moxifloxacin were captured from an electronic database maintained by the manufacturer. Safety data for oral moxifloxacin were obtained from 30 Phase II/III comparator studies (n = 7,368 moxifloxacin, n = 5,687 comparators), 1 Phase IV study (n = 18,374), and 4 postmarketing observational studies (n = 27,756). Safety data for IV moxifloxacin were obtained from 2 Phase III comparator studies (n = 550 maxifloxacin, n = 579 comparators). In addition, pharmacokinetic data were reviewed. RESULTS: In Phase II/III comparator studies, gastrointestinal complaints were the most common adverse drug reactions (ADRs) associated with both formulations of moxifloxacin, with nausea occurring in 7.1% and 3.1% of patients receiving oral and IV moxifloxacin, respectively, and diarrhea occurring in 5.2% and 6.2% of patients. Discontinuation rates due to ADRs with oral and IV moxifloxacin were 2.7% and 6.0%, and mortality rates were 0.3% and 4.0%. Similar rates of withdrawal and mortality were observed in the comparator groups. There was no evidence that moxifloxacin caused disturbances in glucose metabolism in patients with or without diabetes mellitus, and there was no evidence of an increased risk for cardiovascular adverse events. Pharmacokinetic analyses indicated that dose adjustment of moxifloxacin does not appear to be necessary in elderly patients, those with renal dysfunction, or those with mild to moderate hepatic impairment. The pharmacokinetics of moxifloxacin have not been studied in patients with severe hepatic insufficiency. Moxifloxacin does not interact with a number of commonly prescribed drugs, although its absorption is decreased by concomitant administration of iron and cationic antacids. CONCLUSIONS: Based on evidence from >7000 patients in clinical trials and >46,000 patients in postmarketing studies, moxifloxacin is generally well tolerated. Its lack of significant drug interactions in target groups makes it an option in diabetic patients or the elderly, as well as in those with renal impairment. PMID- 15336464 TI - A treatment algorithm for neuropathic pain. AB - BACKGROUND: Neuropathic pain is a chronic pain syndrome caused by drug-, disease , or injury-induced damage or destruction of sensory neurons within the dorsal root ganglia of the peripheral nervous system. Characteristic clinical symptoms include the feeling of pins and needles; burning, shooting, and/or stabbing pain with or without throbbing; and numbness. Neuronal hyperexcitability represents the hallmark cellular mechanism involved in the underlying pathophysiology of neuropathic pain. Although the primary goal is to alleviate pain, clinicians recognize that even the most appropriate treatment strategy may be, at best, only able to reduce pain to a more tolerable level. OBJECTIVE: The purpose of this review is to propose a treatment algorithm for neuropathic pain that health care professionals can logically follow and adapt to the specific needs of each patient. The algorithm is intended to serve as a general guide to assist clinicians in optimizing available therapeutic options. METHODS: A comprehensive review of the literature using the PubMed, MEDLINE, Cochrane, and Toxnet databases was conducted to design and develop a novel treatment algorithm for neuropathic pain that encompasses agents from several drug classes, including antidepressants, antiepileptic drugs, topical antineuralgic agents, narcotics, and analgesics, as well as various treatment options for refractory cases. RESULTS: Any of the agents in the first-line drug classes (tricyclic antidepressants, antiepileptic drugs, topical antineuralgics, analgesics) may be used as a starting point in the treatment of neuropathic pain. If a patient does not respond to treatment with at least 3 different agents within a drug class, agents from a second drug class may be tried. When all first-line options have been exhausted, narcotic analgesics or refractory treatment options may provide some benefit. Patients who do not respond to monotherapy with any of the first- or second-line agents may respond to combination therapy or may be candidates for referral to a pain clinic. Because the techniques used at pain clinics tend to be invasive, referrals to these clinics should be reserved for patients who are truly refractory to all forms of pharmacotherapy. CONCLUSIONS: Neuropathic pain continues to be one of the most difficult pain conditions to treat. With the proposed algorithm, clinicians will have a framework from which to design a pain treatment protocol appropriate for each patient. The algorithm will also help streamline referrals to specialized pain clinics, thereby reducing waiting list times for patients who are truly refractory to traditional pharmacotherapy. PMID- 15336465 TI - Effects of rivastigmine on behavioral and psychological symptoms of dementia in Alzheimer's disease. AB - BACKGROUND: The presence of certain behavioral and psychological symptoms (eg, paranoia, hallucinations, aggression, activity disturbances) in Alzheimer's disease (AD) may predict faster cognitive and functional decline; therefore, such symptoms represent an important treatment target. Behavioral and psychological symptoms of dementia (BPSD) may be caused at least in part by cholinergic deficits. Regulatory studies of rivastigmine in AD were not designed to evaluate effects on BPSD, but further investigation of rivastigmine in AD was prompted by later studies demonstrating behavioral benefits in other types of dementia. OBJECTIVE: The primary aim of this article was to review available data on the behavioral benefits of rivastigmine in patients with AD. METHODS: Relevant data were identified through a MEDLINE search for studies published in peer-reviewed journals through January 2004. The search terms were Alzheimer, behavior, psychosis, and rivastigmine. Data presented at international scientific congresses were also reviewed to ensure that the most recent data were considered. RESULTS: A meta-analysis of three 6-month, placebo-controlled trials of rivastigmine in mild to moderate AD indicated that rivastigmine 6 to 12 mg/d may improve or prevent disruptive BPSD (P < 0.05 vs placebo). In patients with more advanced AD, 2 open-label studies of up to 12 months' duration found that improvements in BPSD were accompanied by a decrease in the use of psychotropic medications. Rivastigmine demonstrated behavioral benefits in patients with dementia with Lewy bodies (DLB) in a double-blind, placebo-controlled study (P < 0.05). In open-label extension studies, rivastigmine provided sustained effects (up to 2 years) in patients with mild to moderate AD or DLB. CONCLUSIONS: The available dats suggest that rivastigmine may be a well-tolerated treatment option for improving or preventing psychotic and nonpsychotic symptoms associated with AD. Prospective, double-blind studies are needed to evaluate these preliminary findings. PMID- 15336466 TI - Peginterferon alfa-2a: a review of approved and investigational uses. AB - BACKGROUND: In October 2002, the US Food and Drug Administration approved peginterferon alfa-2a for the management of chronic hepatitis C virus (HCV) infection. The addition of polyethylene glycol (PEG) moiety to the interferon (IFN) molecule results in a product with altered pharmacokinetic properties. OBJECTIVE: The aim of this article is to review the pharmacology, medications interactions, adverse events (AEs), and approved or investigational uses of PEG IFN alfa-2a for viral hepatitis and oncologic conditions. METHODS: Relevant articles were identified through searches of MEDLINE (1980-July 2003) and EMBASE (1980-July 2003). Search terms included, but were not limited to, peginterferon alfa-2a, pharmacokinetics, pharmacology, pharmacodynamics, and therapeutic use, as well as terms for specific disease states and AEs. Further publications were identified from citations of resulting papers. RESULTS: Pegylation of IFN alfa-2a results in major changes in the pharmacokinetics of the product. Absorption is prolonged and serum concentrations are sustained over the dosing regimen. PEG-IFN alfa-2a has been shown to be more effective with or without ribavirin (RBV), in the management of treatment-naive patients with chronic HCV infection, than unmodified IFN alfa-2a with or without RBV. Results in other disease states are still preliminary. AEs are similar, in incidence and severity, to those occurring with unmodified IFN. They include earlier hematologic symptoms and fewer influenza-like symptoms. Drug-drug interactions are the same as those occurring with the unmodified IFN product. CONCLUSIONS: The pharmacokinetic profile of IFN alfa-2a is improved by pegylation, which enables less frequent administration and results in improved efficacy with a similar side-effect profile. Combination of PEG-IFN alfa-2a with RBV is associated with a greater chance of achieving a sustained virologic response in treatment-naive patients with chronic HCV, compared with unmodified IFN alfa-2a/RBV combinations. Documentation of efficacy in other conditions awaits results of controlled clinical trials. PMID- 15336467 TI - A randomized, double-blind, parallel-group comparison of controlled- and immediate-release oxybutynin chloride in urge urinary incontinence. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of a new PO controlled-release (CR) QD oxybutynin tablet relative to PO immediate-release (IR) TID oxybutynin in patients with urge urinary incontinence (UI). METHODS: In this multicenter, double-blind trial, patients with UI (> or = 7 episode/wk) and frequency (> or = 8 micturitions/d) were randomized to CR or IR oxybutynin for 6 weeks. Patients initiated treatment at 15 mg/d and the dose was adjusted (in 5 mg/d increments) over 2 weeks according to tolerability. Efficacy (UI episodes, voids, absorbent pads used, urgency, and volume voided per micturition) was assessed during the final 2 weeks of treatment. Tolerability was assessed by evaluating adverse events and treatment withdrawals. RESULTS: Of the 125 patients randomized, 94 (75%) were evaluable for efficacy; tolerability was assessed in all patients. In the CR group, 48 patients (91%) were women and 5 (9%) were men; the mean (SD) age was 58.0 (12.4) years (range, 26-78 years). In the IR group, 37 patients (90%) were women and 4 (10%) were men; the mean (SD) age was 60.6 (14.8) years (range, 26-83 years). Both CR and IR oxybutynin significantly reduced the mean number of total UI episodes per week (both P < 0.001 vs baseline). Both treatments produced equivalent reductions in mean voiding frequency and urinary urgency (all P < 0.001 vs. baseline). Significantly more patients rated CR oxybutynin tolerable on the initial dose of 15 mg/d (P = 0.020) and completed the study at a dose of > or = 15 mg/d (P = 0.018). Dry mouth was the most common adverse event, reported by 68% and 72% of patients in the CR and IR oxybutynin groups, respectively. CONCLUSIONS: Among the patients with urge UI included in this study, CR oxybutynin was as effective as IR oxybutynin for improving primary symptoms, with the additional benefit of QD administration. PMID- 15336468 TI - Onset of analgesia and analgesic efficacy of tramadol/acetaminophen and codeine/acetaminophen/ibuprofen in acute postoperative pain: a single-center, single-dose, randomized, active-controlled, parallel-group study in a dental surgery pain model. AB - BACKGROUND: The combination of tramadol and acetaminophen has demonstrated good efficacy in various clinical pain models. However, there is a need for comparisons of the onset of analgesia and other measures of analgesic efficacy with this combination and other strong combination analgesics for the management of acute pain. OBJECTIVE: The goal of this study was to compare the time to onset of analgesia and other measures of analgesic efficacy with tramadol/acetaminophen 75/650 mg (Tr/Ac) and codeine/acetaminophen/ibuprofen 20/500/400 mg (Co/Ac/Ib) in the management of acute pain after oral surgery. METHODS: This was a single center, single-dose, randomized, active-controlled, parallel-group study in healthy subjects who had undergone surgical extraction of > or =1 impacted third molar requiring bone removal. When patients reported at least moderate pain after dental surgery (score > or =5 on a 10-point scale), they were randomized to 1 of 2 treatment groups. The time to onset of analgesia was measured using a 2 stopwatch technique. The time to the onset of perceptible and meaningful pain relief, pain intensity, pain relief, patient's overall assessment, and adverse events were recorded for 6 hours after dosing. RESULTS: One hundred twenty-eight subjects participated in the study, 64 in each treatment group. The 2 groups were similar in terms of baseline pain severity and demographic characteristics (mean age, 23.7 and 23.4 years in the Tr/Ac and Co/Ac/Ib groups, respectively; mean body weight, 58.5 and 60.3 kg). The median times to the onset of perceptible pain relief were a respective 21.0 and 24.4 minutes, and the median times to the onset of meaningful pain relief were 56.4 and 57.3 minutes. Mean total pain relief and the sum of pain intensity difference were also similar in the early period after dosing (0-4 hours). However, between 4 and 6 hours, Co/Ac/Ib was associated with significant differences in both variables compared with Tr/Ac (P < 0.05). Although similar through the 4-hour assessment, mean pain intensity difference was significantly greater with Co/Ac/Ib at 5 to 6 hours. The proportion of patients assessing their assigned treatment as good or better was significantly greater with Co/Ac/Ib compared with Tr/Ac (P < 0.05). The safety profile of Tr/Ac was comparable to that of Co/Ac/Ib. CONCLUSIONS: In this small and selected group of subjects, the onset of analgesia and analgesic efficacy of Tr/Ac was comparable to that of Co/Ac/Ib. Tr/Ac provided rapid and effective analgesia for acute postoperative dental pain in this population. PMID- 15336469 TI - Efficacy of ofloxacin otic solution once daily for 7 days in the treatment of otitis externa: a multicenter, open-label, phase III trial. AB - BACKGROUND: Otitis externa (OE) is an infection of the external auditory canal that is typically treated with topically applied broad-spectrum antibiotics. Twice-daily topical treatment with ofloxacin otic 0.3% solution for 10 days has been reported to be as effective and well tolerated as the standard of care, neomycin sulfate/polymyxin B sulfate/hydrocortisone solution administered 4 times daily for 10 days. OBJECTIVE: This study evaluated the efficacy and safety profile of 7 days of a once-daily regimen of ofloxacin otic 0.3% solution in the treatment of OE. METHODS: This multicenter, open-label, Phase III study was conducted from June 12, 2002, to October 14, 2002. Eligible patients were aged > or = 6 months and had OE of <2 weeks' duration with moderate to severe edema and tenderness involving 1 or both ears and sufficient exudate for microbiologic culture. Ofloxacin otic solution was instilled once daily for 7 days (5 drops for children aged 6 months to <13 years, 10 drops for adolescents/adults aged > or = 13 years). Assessments were conducted at the end-of-treatment visit and 7 to 10 days later (the test-of-cure visit). Medication was supplied free of charge to study participants who incurred no costs for physician visits. RESULTS: Of 489 patients enrolled at 58 sites in 3 countries, 439 were clinically evaluable (173 children, 266 adolescents/adults; 52 % males, 48% females; 47% Hispanic, 45% white; 5% black, and 3% other). The cure rate among clinically evaluable patients was 91% (95% of children, 88% of adolescents/adults); 68% of patients were cured within 7 days. Forty-three potentially pathogenic strains were isolated from 253 microbiologically evaluable patients. Pseudomonas aeruginosa was isolated from 158 (62%) microbiologically evaluable patients and Staphylococcus aureus from 32 (13%). Eradication rates were 96% overall. No serious adverse events were observed. Minor adverse events were experienced by 15 (3%) of 489 patients included in the safety population. The most common adverse events were pruritus (5 patients), increased earache (4 patients), and application-site reactions (3 patients). Overall mean (SD) adherence to therapy was 98% (11.9). CONCLUSIONS: Ofloxacin otic 0.3% solution administered once daily for 7 days was well tolerated and effective in achieving clinical and microbiologic cure of OE. The compliance rates in this study suggests that this regimen may be better accepted by patients than longer, more repetitive regimens. PMID- 15336470 TI - Effects of granisetron in the treatment of nausea and vomiting after laparoscopic cholecystectomy: a dose-ranging study. AB - BACKGROUND: Patients undergoing general anesthesia for laparoscopic cholecystectomy are at high risk for postoperative emetic symptoms (nausea, vomiting, and retching). Antihistamines, butyrophenones, dopamine receptor antagonists, and selective serotonin receptor antagonists (SSRAs) have been investigated for the prevention and treatment of emetic symptoms. However, these drugs are associated with undesirable adverse effects (AEs), such as excessive sedation, hypotension, dry mouth, dysphoria, hallucinations, and extrapyramidal signs. Granisetron hydrochloride is a newer SSRA developed for the prevention and treatment of cytotoxic drug-induced emetic symptoms, but its effects in postoperative laparoscopic cholecystectomy have not been studied. OBJECTIVE: The aims of this study were to assess the efficacy and tolerability of 4 doses of granisetron, and to determine the minimum effective dose, for the control of established emetic symptoms in patients undergoing general anesthesia for laparoscopic cholecystectomy. METHODS: This prospective, randomized, double blind, placebo-controlled, dose-ranging study was conducted in the Departments of Anesthesiology and Surgery, Toride Kyodo General Hospital (Toride, Japan). Male and female patients aged 23 to 68 years with American Society of Anesthesiologists physical status I (no organic, physiologic, biochemical, or psychiatric disturbance) who were experiencing nausea lasting >10 minutes or retching or vomiting within 3 hours after recovery from general anesthesia for laparoscopic cholecystectomy were enrolled. Patients were randomized to receive a single IV dose of placebo or granisetron at 1 of 4 doses (10, 20, 40, or 80 microg/kg). Patients were monitored for 24 hours after study drug administration. RESULTS: One hundred patients (60 women, 40 men; mean [SD] age, 48 [10] years [range, 23-68 years]; mean [SD] height, 158 [7] cm [range, 145-177 cm]; mean [SD] body weight, 56 [8] kg [range, 43-75 kg]) were enrolled. No significant differences in baseline demographic or clinical characteristics were found between the study groups. The proportions of patients who were free of emetic symptoms were significantly higher with granisetron 20, 40, and 80 microg/kg than with placebo (P = 0.02, 0.007, and 0.007, respectively). The difference between the granisetron 10-microg/kg group and the placebo group was not significant. No clinically significant AEs were reported in any group. CONCLUSIONS: Granisetron 20 microg/kg was the minimum effective dose for the treatment of established postoperative emetic symptoms in patients undergoing laparoscopic cholecystectomy. Increasing the dose to 80 microg/kg provided no further benefit. PMID- 15336471 TI - Comparison of cizolirtine citrate and metamizol sodium in the treatment of adult acute renal colic: a randomized, double-blind, clinical pilot study. AB - BACKGROUND: Renal colic causes excruciating pain that provides a good clinical model of acute pain for the development of new analgesics. OBJECTIVE: The purpose of the study was to compare the analgesic efficacy and tolerability of cizolirtine citrate and metamizol sodium in adult acute renal colic. METHODS: This Phase II, randomized, double-blind, clinical pilot study was conducted in the emergency departments of 6 general hospitals in the Czech Republic between October 2000 and February 2001. Male and female patients aged 18 to 65 years and presenting with hematuria and moderate to severe pain due to suspected renal colic starting within the 24 hours before presentation were eligible. Patients were randomized to receive a single IV dose of cizolirtine 350 mg or metamizol 2,500 mg, administered by slow infusion over 15 minutes. Both doses were maximal for the respective drugs to attain adequate analgesia. Use of rescue medication with butorphanol was allowed 30 minutes after study drug administration. Pain intensity was assessed at various time points during the following 360 minutes using a 100-mm visual analog scale (VAS) and a verbal categoric scale. In addition, a specific verbal categoric scale was used to rank pain relief. Physical examinations, laboratory tests, and questioning for adverse events addressed drug tolerability. RESULTS: Sixty-four patients (50 men, 14 women; mean [SD] age, 44.21 [12.29] years; mean [SD] body mass index, 25.97 [3.38] kg/m(2)) were enrolled. Physical examination findings and mean VAS pain intensity scores at baseline (mean [SD], 79.42 [7.89] mm and 82.59 [10.50] mm in the cizolirtine and metamizol groups, respectively) were similar in both groups. After 30 minutes, the mean (SD) scores were 33.84 (25.15) mm and 25.41 (24.51) mm, respectively. This difference was not statistically significant, and the noninferiority of cizolirtine with respect to the comparator could not be established. However, the proportion of patients that showed satisfactory pain relief (ie, decrease of > or = 50% in VAS pain intensity score compared with baseline) at 30 minutes in the cizolirtine group was fairly high (64.5%), which means relevant analgesic activity. Both treatments were well tolerated; 6 adverse events were reported in 5 patients (7.8%), and all were considered as not treatment related. CONCLUSIONS: Although this limited pilot study did not include an internal measure of sensitivity, relevant pain reduction was shown in the population of patients with suspected renal colic undergoing treatment with cizolirtine, suggesting the presence of analgesic activity. However, the efficacy of cizolirtine was found to be similar to that of metamizol. Treatments were well tolerated. PMID- 15336472 TI - Retrospective longitudinal cohort study comparing the effects of angiotensin converting enzyme inhibitors and long-acting calcium channel blockers on total and cardiovascular mortality in patients with hypertension. AB - BACKGROUND: There is controversy regarding the impact that different antihypertensive regimens, including modern combination therapy, have on the incidence of myocardial infarction, other cardiovascular events, and mortality. OBJECTIVE: The objective of this study was to determine and compare the effects of treatment strategies based on angiotensin-converting enzyme (ACE) inhibitors and long-acting calcium channel blockers (CCBs) on total and cardiovascular mortality in hypertensive patients in a usual-care setting. METHODS: This retrospective, longitudinal cohort study used integrated medical and pharmacy claims data from a geographically diverse administrative database of >8 million persons in the United States. Patients aged > or = 18 years with hypertension were eligible if they had filled a prescription for either an ACE inhibitor or a long-acting CCB between January 1, 1995, and June 30, 1999 (the index prescription). Patients who had a prescription for any antihypertensive agents before the index prescription were excluded, as were eligible ACE inhibitor treated patients who used CCBs or CCB-treated patients who used ACE inhibitors during the follow-up period. Use of all other antihypertensive medications was permitted. Patients were matched using a propensity score generated from a logistic regression model. A survival-analysis approach was used to compare mortality between groups. The final cohorts were assessed through June 30, 2002. RESULTS: A total of 18,199 patients met the study inclusion criteria; 12,608 (69.3%) used an ACE inhibitor and 5,591 (30.7%) used a CCB. The mean follow-up was approximately 4.4 years. After cohort matching using the propensity score, the study population consisted of 10,926 patients, 5,463 matched patients in each group. The adjusted hazard ratios (95% CIs) for all-cause, cardiovascular, congestive heart disease, and congestive heart failure mortality in the ACE inhibitor group compared with the CCB group for the entire follow-up period were 0.70 (0.63-0.79), 0.65 (0.53-0.80), 0.47 (0.32-0.70), and 0.74 (0.49-1.12), respectively. CONCLUSION: Analysis of a large medical and pharmacy database suggests that an ACE inhibitor-based treatment strategy is associated with reduced mortality compared with a CCB-based strategy in patients with hypertension in a managed care setting. PMID- 15336473 TI - A randomized, open-label study of the immunogenicity and reactogenicity of three lots of a combined typhoid fever/hepatitis A vaccine in healthy adults. AB - BACKGROUND: Travelers are often advised to receive both the typhoid fever and hepatitis A virus (HAV) vaccines, particularly when going to areas where the 2 diseases are endemic. Thus, combined administration of these vaccines could make immunization more acceptable by reducing the number of injections needed. OBJECTIVE: This study compared the safety profiles and immunogenicity of 3 batches of a combined typhoid fever/HAV vaccine administered using a dual-chamber bypass syringe. METHODS: This randomized, open-label study was conducted at 2 university-based travel clinics in Germany and Austria. Subjects received a single IM injection from 1 of 3 batches of the combined vaccine. Blood samples were drawn immediately before and 28 days after vaccination to evaluate the response to the 2 antigens by assessing geometric mean titers (GMTs) and rates of seroconversion and seroprotection. Subjects recorded all adverse events (AEs) occurring during the study period in a diary. RESULTS: Six hundred ten healthy adults were enrolled in the study. Twenty-eight days after vaccination, 90.6% of the study population had protective typhoid Vi antibody titers (> or = 1 microg/mL) and 100% had protective HAV antibody titers (> or = 20 mIU/mL). Seroconversion rates and GMTs were not significantly different between the 3 batches. There were no differences with regard to local or systemic AEs between the 3 batches of vaccine. There were no immediate adverse reactions (within 30 minutes of vaccination) and no serious AEs related to vaccination. Of 609 evaluable subjects (1 was lost to follow-up after the first visit), 555 (91.1%) experienced > or = 1 local reaction within the first 7 days after vaccination, mainly pain at the injection site (550 [90.3%]), but only 26 (4.3%) described this pain as severe. Vaccine-related headache and mild to moderate asthenia were each reported by 54 subjects (8.9%). Symptoms resolved spontaneously in all cases. CONCLUSIONS: The 3 batches of the combined typhoid fever/HAV vaccine administered by dual-chamber bypass syringe were equally well tolerated and effective in healthy adults, and did not differ significantly in terms of GMTs or seroconversion rates. PMID- 15336474 TI - Association of asthma therapy and Churg-Strauss syndrome: an analysis of postmarketing surveillance data. AB - BACKGROUND: Churg-Strauss syndrome (CSS), also known as allergic granulomatous angiitis (AGA), is a rare vasculitis that occurs in patients with bronchial asthma. The nature of the association of CSS with various asthma therapies is unclear. OBJECTIVE: This study investigated the associations of different multidrug asthma therapy regimens and the reporting of AGA (the preferred code for CSS in the coding dictionary for the Adverse Event Reporting System [AERS]) by applying an iterative method of disproportionally analysis to th AERS database maintained by the US Food and Drug Administration. METHODS: The public-release version of the AERS database was used to identify reports of AGA in patients receiving asthma therapy. Reporting of AGA was examined using iterative disproportionality methods in patients receiving > or =1 of the following drug classes: inhaled corticosteroid (ICS), leukotriene receptor antagonist (LTRA), short-acting beta(2)-agonist (SABA), or long-acting beta(2)-agonist (LABA). The Bayesian data-mining algorithm known as the multi-item gamma poisson shrinker was used to determine the relative reporting rates by calculation of the empirical Bayes geometric mean (EBGM) and its 90% CI (EB05 = lower limit and EB95 = upper limit) for each drug. Subset analyses were performed for each drug with different medication combinations to differentiate the relative reporting of AGA for each. RESULTS: A strong association was found between LTRA use and AGA (EBGM = 104.0, EB05 = 95.0, EB95 = 113.8) that persisted with all combinations of therapy studied. AGA was also associated with the ICS, SABA and LABA classes (EBGM values of 27.8, 14.6 and 40.4, respectively). However, the latter associations were mostly dependent on the presence of concurrent LTRA and, to a lesser extemt, oral corticosteroid therapy and became negligible (ie, EB05 < 2) for patients who were not receiving these concurrent treatments. CONCLUSIONS: Differences based on relative reporting were observed in the patterns of association of AGA with LTRA, ICS, and beta(2)-agonist therapies. A strong association between LTRA use and AGA was present regardless of the use of other asthma drugs. PMID- 15336475 TI - Neuroleptic malignant syndrome associated with ziprasidone in an adolescent. AB - BACKGROUND: Neuroleptic malignant syndrome (NMS) is a rare but potentially fatal disorder characterized by fever, muscular rigidity, delirium, and autonomic instability. Although the classic presentation of NMS has been most commonly associated with the typical neuroleptic medications, sporadic cases in association with atypical neuroleptic medications have been reported. OBJECTIVE: We describe a case report of a pediatric patient with NMS associated with the use of the atypical antipsychotic medication ziprasidone hydrochloride. METHODS: After a MEDLINE search of relevant literature (key terms: atypical antipsychotic, ziprasidone, neuroleptic malignant syndrome, and NMS; years: 1995-2004), no reports of NMS in association with ziprasidone in the pediatric population were identified. RESULTS: The patient was a 15-year-old male adolescent with a history of schizoaffective disorder treated with ziprasidone capsules, 80 mg QD for 8 weeks prior to presentation. He was brought to the emergency department because the family noted that the child had a tactile fever; was rigid, diaphoretic, tremulous, and difficult to arouse; and had persistent urinary incontinence. The patient was admitted to the pediatric intensive care unit, where he remained rigid and unresponsive except for incoherent speech. He was treated for a presumptive diagnosis of NMS with IV dantrolene sodium (2 mg/kg q6h) to reduce the sequele of NMS; urinary alkalinization with sodium bicarbonate to maintain a urinary pH of 6.5 to 7.0; cardiac, pulse oximetry, and vital sign monitoring; and supportive care, including IV saline hydration. CONCLUSION: We present this case to alert physicians of the possibility of NMS in adolescent patients treated with ziprasidone. PMID- 15336476 TI - Evaluation of a low-dose lidocaine iontophoresis system for topical anesthesia in adults and children: a randomized, controlled trial. AB - BACKGROUND: Commonly used classes of topical anesthetics require 30 to 60 minutes to provide effective anesthesia. A new low-dose lidocaine iontophoresis system (LDLIS) may provide topical anesthesia in 10 minutes at a lower dose than previous systems, thereby limiting adverse events. METHODS: This was a prospective, randomized, multicenter, double-blind, placebo-controlled, clinical trial. Adults and children aged 5 to 17 years (inclusive) received a 10-minute ionntophoretic treatment with either lidocaine or a saline placebo before venipuncture or venous cannulation. Intensity of pain associated with venipuncture or venous cannulation was assessed using either a 10-cm Visual Analog Scale (VAS) for adults and children aged 12 to 17 years or the Facial Affective Scale (FAS) for all children enrolled. RESULTS: Five hundred forty eight patients (276 adults, 272 children) participated. Mean (SD) VAS pain scores were lower in adults who received iontophoresis with lidocaine rather than with placebo (0.77 [1.49] vs 2.52 [2.30], P < 0.001) and in children aged 12 to 17 years (1.50 [1.87] vs 2.58 [2.26], P = 0.001). FAS pain scores were lower among children who received iontophoresis with lidocaine rather than with placebo (0.36 [0.26] vs 0.51 [0.27], P < 0.001). Similar results were found for children stratified by age group (5-7 years: 0.40 [0.30] vs 0.60 [0.31], P = 0.011; 8-11 years: 0.35 [0.27] vs 0.48 [0.27], P = 0.021; 12-17 years: 0.33 [0.21] vs 0.48 [0.24], P = 0.001). Mean (SD) parental ratings of pain on the FAS for children aged 5 to 11 years were also lower for the lidocaine group (0.45 [0.28] vs 0.55 [0.25], P = 0.018). Adverse events were similar between groups and included skin erythema and edema. One patient in the study experienced a partial-thickness burn. CONCLUSION: In this study of adults and children, the LDLIS provided effective topical anesthesia for venipuncture and venous cannulation within 10 minutes. PMID- 15336477 TI - Pharmaceutical economics and health policy research using administrative data. PMID- 15336478 TI - Assessing satisfaction with pain medication in primary care patients: development and psychometric validation of a new measure. AB - BACKGROUND: The measurement of patient satisfaction with pain medication (SPM) is a potentially useful aid for health care decision-making, but no validated measures for SPM are known. OBJECTIVE: This study aimed to develop an instrument to assess this patient-reported outcome in primary care and to evaluate whether it satisfactorily fulfilled the required psychometric properties (ie, reliability, validity, and sensitivity to change). METHODS: The measure's content was obtained from literature reviews, focus groups, and expert opinion. A preliminary version of 14 self-administered items was obtained and tested in a prospective study in patients receiving pain medication. Item-total statistics and factor analysis were performed to obtain the final version. The final version was psychometrically validated by assessing feasibility, internal consistency (Cronbach's alpha), convergent validity (multivariant methods), discriminant validity in patients presenting pain relief (receiver operating characteristic curves), test-retest reliability (using the intraclass correlation coefficient [ICC] in patients maintaining medication), and sensitivity to change (in patients changing medication). RESULTS: A total of 1119 patients were recruited (626 women [55.9%]; mean [SD] age, 47.6 [15.36] years; primarily suffering musculoskeletal injury [50.7%]). Four items were deleted from the preliminary version owing to low item-scale correlation and/or factor loadings. The final factor analysis confirmed a 4-factor solution, which explained 81.4% of the variance in questionnaire scores. Adverse events, speed/duration of effect, functional benefit, and overall satisfaction dimensions were identified. Cronbach's alpha and ICC for the 10-item final version were >0.80 for the summary score and all dimensions. Pain-related characteristics (intensity, frequency, and degree of pain relief), but not patient-related characteristics, were independently associated with the summary score. The area under the curve was 0.78 for the summary score. Effect sizes and standardized response mean were > or = 0.84 for the summary score and all dimensions. CONCLUSION: The SPM questionnaire appears to have good acceptability as well as satisfactory psychometric properties, based on these analyses. PMID- 15336479 TI - Are English- and Chinese-language versions of the SF-6D equivalent? A comparison from a population-based study. AB - OBJECTIVE: The goal of this study was to assess the equivalence of English- and Chinese-language versions of the SF-6D (a 6-dimensional health classification system based on the 36-Item Short Form Health Survey) using a model of equivalence proposed previously. METHODS: We analyzed data from a previously published, cross-sectional, population-based survey of ethnic Chinese in Singapore, using linear regression models to adjust for the influence of potential confounding variables. Based on equivalence clinical trial methods, measurement (ie, scale) and item equivalence were assessed by comparing 90% CIs of differences in scores due to language with predefined equivalence margins, that corresponded to the minimum clinically important difference for SF-6D utility and item scores. RESULTS: Data from 2,558 respondents (aged 21-65 years; 48.8% completed the English-language version) were analyzed. The utility scores of respondents using the English- or Chinese-language versions of the SF-6D had similar distribution patterns, with a mean (SD) utility score of 0.8 (0.12). Adjusted 90% CIs for differences in utility and item scores due to language fell within predefined equivalence margins, suggesting measurement and item level equivalence. The 90% CI for scale scores was -0.0089 to 0.0065 (range, 0.0154; equivalence margin, 0.033); the 90% CI for item scores varied from -0.0046 to 0.0020 (range, 0.0026; equivalence margin, 0.0036) for vitality to -0.0024 to 0.0037 (range, 0.0061; equivalence margin, 0.0088) for social functioning. Functional equivalence was suggested because the various aspects of equivalence proposed previously were demonstrated in this study. CONCLUSIONS: English- and Chinese-language versions of the SF-6D demonstrated item, measurement, and functional equivalence in this population-based study comparing 2 widely used languages with very different linguistic structures. This suggests that English and Chinese SF-6D scores can be pooled, thus increasing the representativeness and power of studies using the SF-6D, and providing a basis for studies to value health by obtaining SF-6D utility scores in Asian populations. PMID- 15336480 TI - Management patterns and outcomes of patients with venous thromboembolism in the usual community practice setting. AB - OBJECTIVE: The objectives of this study were to observe a commercially insured sample diagnosed with a venous thromboembolism (VTE) event and treated postevent with warfarin and to detail the thromboembolic and bleeding outcomes in the time periods during warfarin therapy and after discontinuation of such therapy. METHODS: This retrospective, observational cohort study used medical, pharmacy, and eligibility data from 2 US health plans. Study inclusion required an inpatient diagnosis of deep venous thrombosis (DVT) or pulmonary embolism (PE) between January 1, 1998, and December 31, 2000; warfarin, heparin, or low molecular-weight heparin within 30 days after diagnosis; no VTE diagnosis; and no anticoagulant use for 3 months preceding diagnosis. A random sample of medical charts was abstracted to validate VTE events and collect prothrombin time/international normalized ratio (INR) result data. Recurrent VTE events, bleeding events, and proportion of time within INR range were captured in the postindex VTE event time period. Univariate and multivariate statistical techniques were used to assess outcomes. RESULTS: A total of 2,090 patients were identified with a newly diagnosed VTE event (DVT only, 1450; PE with or without DVT, 640). Mean (SD) age was 61.7 (16) years; mean (SD) follow-up time after the index diagnosis was 21.3 (10) months. Overall mean (SD) length of warfarin therapy was 6.6 (6) months. During the follow-up period, 224 patients (10.7%) experienced a recurrent VTE event and 122 patients (5.8%) experienced a bleeding event requiring hospitalization. The cumulative incidence of recurrent VTE events over 3 and 6 months was 9.0% and 10.9%, respectively. Using the chart abstraction subset, patients were within the appropriate INR range 37.7% of the time while receiving warfarin. CONCLUSIONS: Negative outcomes associated with warfarin therapy-recurrent VTE events and bleeding requiring hospitalization-were experienced by 10.7% and 5.8% of patients, respectively. These data suggest that negative outcomes may be more prevalent in usual community medical practice compared with rates observed in the controlled environment of the clinical trial or specialized anticoagulation clinic. PMID- 15336481 TI - Incremental cost-effectiveness of cyclooxygenase 2-selective versus nonselective nonsteroidal anti-inflammatory drugs in a cohort of coumarin users: a pharmacoeconomic analysis linked to a case-control study. AB - BACKGROUND: A previous case-control study involving concomitant users of coumarin and nonsteroidal anti-inflammatory drugs (NSAIDs) found that cyclooxygenase 2 (COX-2)-selective NSAIDs were associated with fewer bleeding complications than nonselective NSAIDs. OBJECTIVE: The goal of this study was to determine the incremental cost-effectiveness of COX-2-selective versus nonselective NSAIDs in relation to the occurrence of bleeding complications in a cohort of concomitant coumarin users. METHODS: The pharmacoeconomic evaluation was linked to a case control analysis (patients with and without bleeding complications) based on data from the earlier study in users of concomitant coumarin and NSAIDs. Medical costs associated with NSAID use and bleeding complications were estimated according to Dutch guidelines for pharmacoeconomic analyses, based on Dutch drug prices and national averages for health care costs. Rofecoxib, meloxicam, and nabumetone were considered COX-2 selective. Total costs were calculated and compared for 2 hypothetical scenarios in which patients used either COX-2-selective or nonselective NSAIDs. Sensitivity analyses were performed in which both the odds ratios (ORs) and the costs of NSAIDs and bleeding episodes were varied. RESULTS: A total of 1,491 bleeding complications occurred in 4400 coumarin users: among the 221 (15%) NSAID users with a bleeding episode, 96% used a nonselective NSAID and 4% used a COX-2-selective NSAID. The adjusted OR of a bleeding episode for nonselective compared with COX-2-selective NSAIDs was 3.07 (95% CI, 1.18-8.03). The estimated mean cost of a bleeding episode was 478 per patient. Factoring in the excess cost of a COX-2-selective NSAID compared with a nonselective NSAID, as well as the cost savings in averted bleeding episodes, it was determined that there would be net medical cost savings of 53,800 and 162 averted bleeding episodes if the entire patient group received COX-2-selective NSAIDs rather than nonselective NSAIDs. The sensitivity analysis showed these results to be robust. CONCLUSION: In this study population of concomitant coumarin and NSAID users, the reduction in bleeding complications with the use of more expensive COX-2 selective inhibitors was associated with net medical cost savings compared with nonselective NSAIDs. PMID- 15336482 TI - A cost threshold analysis of ciprofloxacin-dexamethasone versus ofloxacin for acute otitis media in pediatric patients with tympanostomy tubes. AB - OBJECTIVE: The aim of this study was to determine the cost-effectiveness hreshold of a ciprofloxacin 0.3% and dexamethasone 0.1% (CD) otic suspension relative to olfloxacin otic solution (OFX) for the treatment of acute otitis media in pediatric patients with tympanostomy tubes (AOMT). METHODS: This study used a decision-analytic model to simulate the costs and consequences of the ototopical treatment of AOMT. The AOMT model consisted of 3 tiers of antimicrobial therapy. Each successive tier represented the repeat treatment of clinical failures from the preceding tier. Patients were modeled for treatment until cured or until third-tier therapy was complete, at which time patients were considered cured. First-tier therapy modeled a comparison of CD and OFX using efficacy rates taken from a randomized clinical trial with a population of 599 patients. Second-tier therapy modeled the use of amoxicillin and clavulanic acid using an efficacy rate taken from the medical literature. Third-tier therapy was modeled as being pathogen specific and could follow 1 of 3 possible clinical pathways: (1) PO fluconazole, (2) IM ceftriaxone, or (3) IV antibiotics administered in a hospital setting. Third-tier therapeutic pathway probabilities were based on the microbiologic spectrum of the treatment failures from the clinical trial. Cost information (in year-2003 US dollars) was taken from accepted cost reference sources and presented from the perspective of a third-party payer. The economic outcome of interest was the cost-effectiveness threshold of CD relative to OFX. RESULTS: Given the model parameters, CD had a cost-effectiveness threshold value of 4.5 times the wholesale acquisition cost of OFX. Based on actual cost, first tier CD therapy was more cost-effective than OFX up to a threshold price of US 152.64 dollars. CONCLUSION: In this decision-analytic model, CD was more cost effective than OFX for AOMT therapy in pediatric patients up to a threshold price of 4.5 times the price of OFX. PMID- 15336483 TI - Insulin glargine clinical trials. PMID- 15336485 TI - Twenty years of the pPS10 replicon: insights on the molecular mechanism for the activation of DNA replication in iteron-containing bacterial plasmids. AB - This review focuses on the contributions of the Pseudomonas replicon pPS10 to understanding the initiation of DNA replication in iteron-containing plasmids from Gram-negative bacteria. Dimers of the pPS10 initiator protein (RepA) repress repA transcription by binding to the two halves of an inverted repeat operator. RepA monomers are the active initiator species that bind to four directly repeated sequences (iterons). pPS10 initiator was the first Rep protein whose domains were defined (two "winged-helix," WH modules) and their binding sites were identified at each half of the iteron repeat. This was confirmed by the crystal structure of the monomer of a homologous initiator (RepE from F plasmid) bound to iteron DNA. The recently solved structure of the dimeric N-terminal domain (WH1) of pPS10 RepA, when compared to the RepE monomer, shows that upon dimer dissociation an alpha-helix at WH1 C-terminus becomes part of an interdomain beta-sheet. In solution, the iteron sequence, by itself, can induce the same kind of structural transformation in RepA. This seems to alter the package of both WH domains to adapt their DNA reading heads (HTH motifs) to the distinct spacing between half repeats in iterons and operator. Based on biochemical and spectroscopic work, structural and functional similarities were proposed between RepA and archaeal/eukaryal initiators. This was independently confirmed by the crystal structure of the archaeal initiator Cdc6. Characterization of mutants, either in pPS10 or in the Escherichia coli chromosome, has provided some evidence on a WH1-mediated interaction between RepA and the chromosomal initiator DnaA that results in a broadened-host range. PMID- 15336486 TI - Characterization of pC7 from Lactobacillus paraplantarum C7 derived from Kimchi and development of lactic acid bacteria--Escherichia coli shuttle vector. AB - A cryptic plasmid pC7 was characterized, which was isolated from Lactobacillus paraplantarum C7 derived from Kimchi, a traditional korean vegetable food. The plasmid pC7 is a circular molecule of 2,134 base-pairs in length with a G + C content of 38.5%. The nucleotide sequence analysis revealed the presence of an open reading frame encoding a putative 317 amino acids protein homologous to replication proteins RepA. Furthermore, a putative double and a single-strand origin were identified. Together with features of nucleotide sequences, the detection of single-stranded intermediate DNA in Lb. paraplantarum demonstrated that pC7 replicates via a rolling circle mechanism. A cloning vector for lactic acid bacteria was developed on the basis of the pC7 replicon, into which were inserted an erythromycin resistance gene as a marker, multiple cloning sites, and Escherichia coli ColE1 replication origin. E. coli and several species of Lactobacillus and Leuconostoc can be transformed with the resultant vector. Therefore, pC7 derivatives may be useful LAB-E. coli shuttle vectors, which are essential in engineering important strains in food fermentation. PMID- 15336487 TI - Molecular analysis of oriT and MobA protein in the 7.4 kb mobilizable beta lactamase plasmid pSJ7.4 from Neisseria gonorrhoeae. AB - The mobilization region of the 7.4 kb beta-lactamase plasmid pSJ7.4 from Neisseria gonorrhoeae was characterized. The 3.2 kb HindIII-BamHI fragment of pSJ7.4 was mobilized between Escherichia coli strains by conjugative plasmid RK2. Selected restriction enzyme-generated deletions of this fragment were subcloned in pACYC177 to obtain constructs that were suitable for analysis of the mobilization region. Mating experiments showed that a highly conserved 1.9 kb DNA region within coordinates 4096-5997 is required for mobilization of pSJ7.4. This region contains two genes encoding the mobilization protein MobA, another protein, MobC, putatively involved in mobilization from some hosts, and an intergenic oriT. The 168 bp intergenic sequence also contains the promoters for mobA and mobC in an arrangement that suggests divergent transcription and autoregulation from oriT. The 56 kDa MobA was expressed in E. coli as a (6x)His Tag fusion protein. Purified MobA specifically induced plasmid relaxation by nicking at the oriT. MobA is exceptional because the N-terminal region alone can mobilize pSJ7.4, albeit at a lower frequency than the full-length protein, even in the absence of MobC. The carboxyl terminal region of MobA did not share homology with other mobilization proteins, but may be involved in promoting efficient transfer of pSJ7.4. PMID- 15336488 TI - Comparative genomics identified two conserved DNA modules in a corynebacterial plasmid family present in clinical isolates of the opportunistic human pathogen Corynebacterium jeikeium. AB - Investigation of 62 clinical isolates of the opportunistic human pathogen Corynebacterium jeikeium revealed that 17 possessed plasmids ranging in size from 7.6 to 14.9 kb. The plasmids formed four groups on DNA restriction analysis. The complete nucleotide sequence of a representative from each group (pK43, pK64, pCJ84, and pB85766) was subsequently determined. Additionally, two plasmids (pCo455 and pCo420) were shown to be derivatives of pK43 and pK64 carrying insertion sequences of the IS3 family. Comparative genomics identified a conserved plasmid backbone consisting of two distinct DNA modules. Conserved motifs in the parAB-repA module indicated that the sequenced plasmids from C. jeikeium are new members of the pNG2 family. Recombinant derivatives of pK43 were shown to replicate in the soil bacterium Corynebacterium glutamicum and in the human pathogen Corynebacterium diphtheriae. The second plasmid module most likely encodes a novel type of DNA invertase. The respective gene is flanked by highly conserved 112-bp inverted repeats. All plasmids are 'loaded' with a characteristic set of genes encoding products of unknown function. Plasmids indistinguishable from pK43 by DNA restriction analysis were identified in different C. jeikeium strains, which revealed 16S-23S rDNA spacer length polymorphisms and specific antibiotic susceptibility profiles, implying a wide dissemination of the plasmid in clinical isolates of C. jeikeium. PMID- 15336489 TI - Factors required in vitro for excision of the Bacteroides conjugative transposon, CTnDOT. AB - Four genes have been found to be essential for excision of the Bacteroides conjugative transposon CTnDOT in vivo: intDOT, orf2c, orf2d, and exc. The intDOT gene encodes an integrase that is essential for integration and excision. The function of the other genes is still uncertain. Previously, we developed an in vitro system for the integration reaction. We have now developed an in vitro system for excision. In this system, the left and right junctions of CTnDOT, attL, and attR, are provided on separate plasmids. The excision reaction produced a cointegrate which contained the attDOT (the joined ends of CTnDOT) and attB (the chromosomal target site). Cointegrate formation was observed after electroporation of Escherichia coli with the assay mixture and was also detected directly in the assay mixture by Southern hybridization. The highest reaction frequencies (10(-3)) were obtained with a mixture that contained purified IntDOT and a cell extract from Bacteroides thetaiotaomicron 4001, which contained the excision region of CTnDOT carried on a plasmid. An unexpected finding was that the addition of purified Exc, which is essential for excision in vivo, was not required for excision in vitro, nor did it increase the frequency of cointegrate formation. PMID- 15336490 TI - Cloning, sequencing, and sequence analysis of two novel plasmids from the thermophilic anaerobic bacterium Anaerocellum thermophilum. AB - The nucleotide sequence of two novel plasmids isolated from the extreme thermophilic anaerobic bacterium Anaerocellum thermophilum DSM6725 (A. thermophilum), growing optimally at 70 degrees C, has been determined. pBAS2 was found to be a 3653 bp plasmid with a GC content of 43%, and the sequence revealed 10 open reading frames (ORFs). The two largest of these, namely Orf21 and Orf41, showed similarity to a Bacillus plasmid recombinase and a Pseudoalteromonas plasmid replication protein, respectively. A sequence with homology to double stranded replication origins from rolling circle plasmids was found, but no single stranded intermediates, characteristic of rolling circle replication, were found on Southern blots. The larger plasmid, pBAL, was found to be a 8294 bp plasmid with a GC content of 39%. It revealed 17 ORFs, of which three showed similarity at the amino acid (aa) level to known proteins. Orf22 showed the strongest similarity (33% aa) to replication proteins from large multiresistance Staphylococcal and Lactococcal plasmids, all of which are believed to replicate via a theta-like replication mechanism. Orf32 showed similarity to both DNA repair proteins and DNA polymerases with highest similarity to DNA repair protein from Campylobacter jejuni (25% aa). Orf34 showed similarity to sigma factors with highest similarity (28% aa) to the sporulation specific Sigma factor, Sigma 28(K) from Bacillus thuringiensis. PMID- 15336491 TI - A rat model of radiofrequency ablation of trigeminal innervation via a ventral approach with stereotaxic surgery. AB - Neurotrophic keratopathy (NK), a consequence of sensory denervation of the cornea, must be better understood in order to develop new approaches to therapy. The purpose of this study was to create a rat model for neurotrophic keratopathy by denervating the trigeminal nerve through a ventral approach with stereotaxic surgery. Stereotaxic coordinates were measured in 46 male Sprague Dawley rat cadavers for localization of V1. After further refining the coordinates in nine live animals, radiofrequency ablation was chosen as an effective method of disrupting the innervation to the cornea. Fifty-two live rats were treated with radiofrequency ablation to define the anatomical localization of the lesion by utilizing gross and histopathological studies. A gross lesion of the trigeminal nerve and/or ganglion was observed in 47 (90%) of the 52 animals. Histopathological studies revealed that all 52 animals had anatomical damage of the trigeminal innervation to the eye. Low mortality and little morbidity were observed in these animals. We have developed a rat model for neurotrophic keratopathy that is simple to produce, accurate in creating a lesion by utilizing stereotaxic techniques combined with radiofrequency ablation, and successful in decreasing morbidity and mortality. PMID- 15336492 TI - Expression and activation of the epidermal growth factor receptor in differentiating cells of the developing and post-hatching chicken lens. AB - The epidermal growth factor receptor is hypothesized to play an important role in the post-natal growth and differentiation of the ocular lens. Immunohistochemistry and western blotting were utilized to examine the distribution and activation of the epidermal growth factor receptor in embryonic and post-hatching chicken lenses. Although present at constant levels within epithelial cells throughout embryonic development, the receptor becomes increasingly activated on a highly conserved tyrosine residue necessary for intracellular signal transduction as hatching approaches. After hatching, activated receptors are found in epithelial cells committed to fiber cell formation and in fiber cells undergoing initial stages of terminal differentiation. Activated receptors could not be identified in central epithelial cells or nuclear fiber cells. This pattern persists until at least one year post-hatching. These data indicate that the epidermal growth factor receptor is positioned to influence not only post-natal patterns of lenticular gene expression but also the greatest amount of lens growth and development. PMID- 15336493 TI - A mechanistic study of the photooxidation of A2E, a component of human retinal lipofuscin. AB - A major constituent of human retinal lipofuscin is A2E (2-[2,6-dimethyl-8-(2,6,6 trimethyl-1-cyclohexen-1-yl)-1E,3E,5E,7E-octatetraenyl]-1-(2-hydroxyethyl)-4-[4 methyl-6(2,6,6-trimethyl-1-cyclohexen-1-yl)-1E,3E,5E,7E-hexatrienyl]-pyridinium). Light transmitted by the lens is absorbed by A2E and the processes initiated by this absorption has been implicated in several maculopothies. The purpose of this study was to evaluate the dominant photochemical mechanisms involved in these reactions, whether through free radical or singlet oxygen intermediacy. The photodestruction of A2E occurs faster in water vs. chloroform and hydrogenated vs. perdeuterated methanol. Both results suggest a free radical mechanism. Product distributions indicate sequential oxygen addition rather than the addition of two oxygen atoms which would be expected if singlet oxygen was an intermediate. Finally, EPR trapping studies lead to the detection of superoxide as the primary intermediate in the photochemical reactions. It is concluded that if singlet oxygen is involved in these photochemical processes it is of minor importance. PMID- 15336494 TI - Localization and regulation of glucagon receptors in the chick eye and preproglucagon and glucagon receptor expression in the mouse eye. AB - Myopia is a condition in which the eye is too long for the focal length of cornea and lens. Analysis of the messengers that are released by the retina to control axial eye growth in the animal model of the chicken revealed that glucagon immunoreactive amacrine cells are involved in the retinal image processing that controls the growth of the sclera. It was found that the amount of retinal glucagon mRNA increased during treatment with positive lenses and pharmacological studies supported the idea that glucagon may act as a stop signal for eye growth. Glucagon exerts its regulatory effects by binding to a single type of glucagon receptor. In this study, we have sequenced the chicken glucagon receptor and compared its DNA and amino acid sequence with the human and mouse homologues. After sequencing about 80% of the receptor, we found a homology between 79.4 and 75.6% on cDNA level. At the protein level, about 73% of the amino acids were identical. Moreover, the cellular localization and regulation of the glucagon receptor in the chick retina was studied. In situ hybridization studies showed that many cells in the ganglion cell layer and inner nuclear layer, and some cells in the outer nuclear layer, express the receptor mRNA. Injection of the glucagon agonist Lys17,18,Glu21-glucagon induced a down-regulation of glucagon receptor mRNA content. Since the mouse would be an attractive mammalian model to study the biochemical and genetic basis of myopia, and because recent studies have demonstrated that form deprivation myopia can be induced, the expression of preproglucagon and glucagon receptor genes were also studied in the mouse retina and were found to be expressed. PMID- 15336495 TI - Restoration of visual responses following transplantation of intact retinal sheets in rd mice. AB - PURPOSE: To correlate the functional outcomes with histologic findings following transplantation of fetal retinal sheets in rd mice, and to investigate the mechanisms of visual function restoration. METHODS: Twenty-one postnatal day 31 38 rd/rd (C3H/HeJ) mice were transplanted in one eye with retinal sheets (1.0 x 0.4 mm) obtained from embryonic day (E) 17 enhanced-green-fluorescent protein (eGFP) mice. Five mice underwent sham surgery without insertion of tissue. Four to five weeks after transplantation, visual responses to a light flash were recorded across the superior colliculus (SC) in seven eyes of seven transplanted mice that had clear corneas and lenses, and in all five sham surgery mice. Following the SC recording, the eyes were enucleated and processed for immunohistochemistry and examined using confocal microscopy. RESULTS: In three out of the seven eyes (43%), positive responses were recorded in the SC in an area topographically corresponding to the placement of the transplant in the host retina. No responses were recorded in the untreated eyes of 5-week-old and 9-week old rd/rd mice, and in the 9-week-old sham surgery mice. In contrast, visual responses were recorded over the entire SC in normal eyes. The response onset latencies of the 3 transplanted mice with responses were similar to those of normal control mice. The organization of the graft did not appear to correlate as expected with the electrophysiology results, as eyes with well-organized, laminated grafts showed no response whereas the three light-responsive eyes had rosetted or disorganized grafts. All three light-responsive eyes demonstrated much higher levels of recoverin immunoreactivity in the host retina overlying the graft compared with untreated age-matched rd/rd mice. CONCLUSION: Restoration of the SC visual response does not appear to depend on a well-organized transplant in the rd mouse. Increased recoverin-staining in the host retina in light responsive animals suggested that host cone rescue was the likely mechanism of vision restoration in this transplant model. PMID- 15336496 TI - Tensile mechanical and creep properties of Descemet's membrane and lens capsule. AB - Descemet's membrane (DM) and the lens capsule (LC) are two ocular basement membrane structures which in comparison with other basement membranes have exceptional thicknesses which increase with age. Both membranes are supposed to contain networks of type IV collagen and laminin linked together with nidogen/entactin and containing other glycoproteins and proteoglycans. DM is a unique basement membrane which in addition contains fine filaments of type VIII collagen arranged in a hexagonal lattice. The mechanical functions of the LC are in lens suspension and accommodation, and its mechanical properties, previously investigated, are of great interest from a surgical point of view. DM serves as an endothelial basement membrane. Otherwise, its physiological function is unknown but may be one of mechanical support, filtration, or fluid barrier. Data on the mechanical properties of DM or the supramolecular assembly of type VIII collagen are very scarce or absent. The aim of this study was to determine and compare the mechanical properties of the two ocular membranes in order to elucidate the properties of DM in the light of those of LC. The human eyes were from testamentary donors and rat, cow, and sow eyes were obtained from normal animals. The tensile mechanical properties were determined by a volume-strain procedure and creep properties by subjecting the membranes from the latter three species to a constant axial stress. In rat, cow, and sow, DM was less strained to obtain a fixed moderate stress value (0.5 MPa) and showed to be 3.4- to 5.2-fold stiffer and to attain 2.7- to 4.6-fold higher stress at a strain value of 0.10 when compared with LC. The maximal strain, stiffness and stress were found to be less than those of the LC. In humans, DM and LC showed very similar mechanical properties. The instantaneous creep of DM was found to be less than that of LC indicating a higher stiffness of DM in the axial direction. In conclusion, depending on the species, DM showed to possess from one-fourth to similar mechanical strength as that of LC, and, in rat, cow, and sow, DM appeared to be stiffer in both lateral and axial directions at moderate strain values when compared with LC. PMID- 15336497 TI - Developmental changes of aggrecan, versican and neurocan in the retina and optic nerve. AB - We have used a monoclonal antibody to neurocan and specific polyclonal antibodies to the non-homologous glycosaminoglycan attachment regions of aggrecan and mRNA splice variants of versican to compare the localization and developmental changes of these structurally related hyaluronan-binding chondroitin sulfate proteoglycans in the rat retina and optic nerve. Staining for aggrecan and versican was first seen at embryonic day 16 in the optic nerve and retina, whereas neurocan was not detected in the embryonic eye. At postnatal day 0 (P0), beta-versican staining is largely confined to the inner plexiform layer whereas alpha-versican is also apparent in the neuroblastic layer. Both aggrecan and, much more weakly, neurocan immunoreactivity is present throughout the neonatal retina. At P9, aggrecan and versican immunoreactivity is most intense in the inner and outer plexiform and ganglion cell layers, accompanied by diffuse staining in the inner and outer nuclear layers. Aggrecan and alpha-versican are also present throughout the optic nerve and disk, whereas beta-versican and neurocan are confined to the laminar beams of the optic nerve. Between P0 and P9 there is a marked increase in beta-versican expression in the inner and outer nuclear layers and in the outer plexiform layer, whereas there is only weak staining of neurocan in the inner plexiform and ganglion cell layers of P9 retina. By 1 month postnatal the staining pattern of the fully differentiated retinal layers is essentially identical to that seen in the adult, where there is strong aggrecan and alpha-versican immunoreactivity in the retina and optic nerve, whereas beta-versican has essentially disappeared from the adult retina and, similarly to neurocan, is present only in the laminar beams of the optic nerve. The marked decrease of beta-versican in the retina is consistent with >90% decrease in its concentration in brain during postnatal development, suggesting that the developmental time-course for these proteoglycans in retina parallels that seen in other areas of the central nervous system. PMID- 15336498 TI - Etanercept treatment in the endotoxin-induced uveitis of rats. AB - This study was conducted to investigate therapeutic value of a soluble tumor necrosis factor-alpha (TNF-alpha) receptor, etanercept, in a rat model of endotoxin-induced uveitis (EIU). Forty-two inbred male Lewis rats were divided into seven equal groups. 200 microg of Escherichia coli 055:B55 lipopolysaccharide (LPS) was injected in one hind footpad of the Groups 2, 3, 4, 5, 6, and 7 rats. Group 5, 6, and 7 rats also received subcutaneous etanercept 24 hr prior to LPS injection at a dose of 0.4 mg kg(-1). Group 1 rats were used as controls. Eight, 24, and 48 hr after treatment clinical uveitis scores (miosis, iris hyperemia, and hypopyon) were assessed by a masked observer and the rats were euthanized. Neutrophil leukocytes, CD8+, CD4+, and CD45RO+ cells in the anterior uveal tissue were counted either after hematoxylin-eosin or monoclonal antibody staining. TNF-alpha levels were also measured in the aqueous humor samples by an ELISA method. Etanercept treatment significantly improved clinical uveitis scores at all examination points compared to the LPS injected animals. The improvement was almost complete expect for the miosis score, since no significant difference was detected between the controls and LPS + Etanercept treated animals at all examination points. Cell counts were also at significantly lower levels in LPS + Etanercept treated animals at all examination points, except for CD8+ and CD45RO+ cell counts at 24 hr examination point. There was no significant difference between the controls and LPS + Etanercept treated animals at all examination points as with CD4+ and CD45RO+ cell counts at 48 hr. Our data showed that etanercept had a definite effect on the treatment of EIU. Further studies should clarify its efficacy on clinical uveitis conditions. PMID- 15336499 TI - Damaged DNA-binding protein 2 accelerates UV-damaged DNA repair in human corneal endothelium. AB - PURPOSE: To determine damaged DNA-binding protein 2-gene expression levels in vitro and ex vivo, and the degree of DNA repair in damaged DNA-binding protein 2 overexpressing cultured human corneal endothelium after ultraviolet irradiation. METHODS: Constitutive damaged DNA-binding protein 2-gene expression levels in various human tissues were determined by semi-quantitative reverse transcription polymerase chain reactions. The dynamics of nucleotide excision repair-related gene expression in cultured human corneal endothelium were investigated in a ribonuclease protection assay after ultraviolet-irradiation. The effect of damaged DNA-binding protein 2 on DNA repair was studied after ultraviolet irradiation in cultured human corneal endothelium infected with adenovirus carrying damaged DNA-binding protein 2. RESULTS: Human corneal endothelium and epithelium in the donor cornea had the highest constitutive damaged DNA-binding protein 2-gene expression of the various human tissues studied. Gene expression level dynamics associated with nucleotide excision repair factors after ultraviolet-irradiation showed that the increase in the rate of damaged DNA binding protein 2-gene expression in cultured human corneal endothelium was highest of the nucleotide excision repair-related genes studied. An in vivo DNA repair assay showed that DNA repair efficiency in damaged DNA-binding protein 2 overexpressing cultured human corneal endothelium after ultraviolet-irradiation was significantly improved as compared with that in the control human corneal endothelium. CONCLUSION: The human corneal endothelium abundantly expresses the damaged DNA-binding protein 2-gene that is produced efficiently on ultraviolet exposure. This overexpressed damaged DNA-binding protein 2 in the human corneal endothelium contributes to the protection system against DNA damage after ultraviolet-irradiation. Our findings show a critical role for damaged DNA binding protein 2 in DNA repair to maintain the human corneal endothelium function. PMID- 15336500 TI - Beta-crystallin association. AB - Beta-crystallins are major protein constituents of the mammalian lens, where their stability and association into higher order complexes are critical for lens clarity and refraction. Dimerization is an initial step in formation of beta crystallin complexes. Beta-crystallin association into dimers is energetically highly favoured, but rapidly reversible under physiological conditions. Beta crystallin dimers can exchange monomers, probably through a transient and energetically unfavoured monomer intermediate state. As predicted by molecular modelling, the fraction of beta-crystallin present as dimers increases with increasing temperature, implying that beta-crystallin association is entropically driven. PMID- 15336501 TI - Identification of protein phosphatase 2C and confirmation of other protein phosphatases in the ocular lenses. AB - The reversible phosphorylation of proteins plays essential roles in regulating various cellular events, and is regulated by the opposing actions of protein kinases and protein phosphatases. Protein kinases in the lens system have been well studied, but very little is known about lens protein phosphatases. Protein phosphatases can be divided several families, such as protein phosphatase types 1, 2A, 2B and 2C (PP1, PP2A, PP2B and PP2C) and protein tyrosine phosphatases (PTP). In this study we evaluated what kinds of protein phosphatases are present in the lens by using various specific substrates and inhibitors. Samples were prepared from lenses of 17-day-old chick embryos, and fractionated by high resolution gel permeation column chromatography, then the fractions were assayed for phosphatase activities. The results with 32P-labeled glycogen phosphorylase A, okadaic acid and inhibitor-1, which are a specific substrate and inhibitors of PP1 and/or PP2A, showed that PP1activities were present in the 500-, 115- and 45 kDa fractions of the lens protein. The 115-kDa fraction also contained PP2A activity. By using a phosphothreonine-containing peptide as a substrate, three peaks of phosphatase activities were found at around 115, 55 and 35 kDa. Based on their response to various phosphatase inhibitors and their metal dependency, the fractions of 115 and 35 kDa were concluded to contain PP2A, while the 55-kDa fraction contained PP2C. Immunoblot using specific antibodies against PP1, PP2A and PP2C confirmed that each fraction above contained corresponding protein phosphatases as proteins. When a phosphotyrosine-containing peptide substrate was examined at pH 7.4, we observed a major peak at 500 kDa, which was presumed to contain receptor-like PTP(s). On the other hand, at pH 5.5, we observed a peak of 18 kDa, which was confirmed to contain a low-molecular-weight PTP. These protein phosphatases have recently been suggested to be involved in stress response and apoptosis. Their physiological roles in the lens are of much interest. PMID- 15336502 TI - Human alphaA- and alphaB-crystallins prevent UVA-induced apoptosis through regulation of PKCalpha, RAF/MEK/ERK and AKT signaling pathways. AB - AlphaA- and alphaB-crystallins are distinct antiapoptotic regulators. Regarding the antiapoptotic mechanisms, we have previously demonstrated that under staurosporine treatment, HalphaA- and HalphaB-crystallins can interact with Bax and Bcl-XS, proapoptotic members of the Bcl-2 family, to sequester their translocation into mitochondria, and thus prevent the staurosporine-induced apoptosis. In the present study, we further compared the anti-apoptotic mechanisms of HalphaA- and HalphaB-crystallin in preventing human lens epithelial cells from UVA-induced apoptosis. UVA-irradiation of human lens epithelial cells turned on the apoptotic death program. Moreover, associated with the activation of the death program, UVA also activated the RAF/MEK/ERK signaling pathway. In contrast, p38 kinase and JNK1/2 signaling pathways were not activated. Inhibition of the RAF/MEK/ERK pathway by a dominant negative mutant RAF1 greatly attenuated UVA-induced apoptosis. Expression of the exogenous human alphaB-crystallin prevented UVA-induced activation of RAF/MEK/ERK pathway and thus substantially abrogated UVA-induced apoptosis. In contrast, expression of the exogenous human alphaA-crystallin did not prevent UVA-induced activation of RAF/MEK/ERK pathway. Instead, it activated AKT kinase pathway to promote survival and thus counteracted the UVA-induced apoptosis. Together, our results for the first time reveal that by regulating multiple signaling pathways the two alpha-crystallins can prevent stress-induced apoptosis through different mechanisms. PMID- 15336503 TI - Effects of matrix metalloproteinase-3 gene transfer by electroporation in glaucoma filter surgery. AB - To develop gene therapy that can be applied to glaucoma-filtering surgery, we studied effects of transfection of matrix metalloproteinase-3 (MMP-3) cDNA into rabbit conjunctiva by electroporation (EP) on changes of intraocular pressure (IOP) and bleb formation after glaucoma filtering surgery. pTracer-CMV2 vector containing MMP-3 cDNA was transfected into rabbit conjunctiva by EP and MMP-3 expression was studied by reverse transcription (RT)-PCR, zymography and western blot analysis. Three days after the EP transfection of MMP-3 cDNA or vector alone into rabbit conjunctiva, trabeculectomy was performed at the place of transfection in the presence or absence of 0.04% mitomycin C (MMC). Then changes in IOPs and bleb formation were compared with each other. Expression of MMP-3 was detected in conjunctiva until 30 days after transfection by EP. Trabeculectomy following MMP-3 transfection caused significantly longer survival of filtering bleb and decreased levels of IOP in comparison with controls (trabeculectomy alone or trabeculectomy following vector transfection), and these levels were almost identical to those of trabeculectomy with MMC. The present study indicates that EP is effective to transfect some genes that promote the filtering bleb formation in glaucoma surgery, such as MMP-3 gene, and this may be potentially applicable to glaucoma-filtering surgery in glaucoma patients. PMID- 15336504 TI - Comparison of characteristics of peroxide-conditioned immortal human lens epithelial cell lines with their murine counterparts. AB - Previously, this laboratory has reported the characteristics of murine immortal lens-epithelial cells (alphaTN4-1) conditioned to survive either H2O2 or tertiary butyl hydroperoxide (TBOOH) stress. This communication now describes similar observations upon human HLE-B3 cells. It was found that the human cells are more sensitive to peroxides than their murine counterpart. Similar to the murine cells, conditioning to TBOOH endows the HLE-B3 cells with resistance to H2O2 but unlike the murine cells, conditioning to H2O2 gives the human cells resistance to TBOOH. Furthermore, while withdrawal of TBOOH stress from TBOOH-conditioned alphaTN4-1 cells causes a loss of resistance to this peroxide but not H2O2, with human cells resistance to both peroxides is retained. Examination of the antioxidative defense (AOD) enzyme activities show an extraordinary increase in catalase activity and significant augmentation of most other enzymes assayed in all conditioned human cell lines. In contrast, it was previously found that only catalase and glutathione-S-transferase have considerable increases in activity in the murine lines. However, in most cases, the AOD enzyme activity in murine control cells is about 2-fold higher than in human control cells. The gene expression of human TBOOH-conditioned (Thum) and control (Chum) lines were also examined utilizing microarray analysis. Surprisingly, no significant change in gene expression was found for any of the prominent AOD enzymes. Such results differ from the response of murine cells where many AOD enzymes have increased expression. These observations suggest while the same AOD enzymes may be utilized in both murine and human lens-epithelial cells, the levels at which they are maintained and the manner in which they are recruited in response to stress may differ. PMID- 15336505 TI - Support for a proposed retinoid-processing protein complex in apical retinal pigment epithelium. AB - The interaction of cellular retinaldehyde-binding protein (CRALBP) with ERM (ezrin, radixin, moesin)-binding phosphoprotein 50 (EBP50) in retinal pigment epithelium (RPE) microsomes has led to the hypothesis that a retinoid-processing protein complex exists in apical RPE. Mouse RPE apical processes were isolated on wheat germ agglutinin-coated agarose beads. Proteomic analyses of the isolated apical RPE demonstrated the presence of CRALBP, EBP50, 11-cis-retinol dehydrogenase, cellular retinol-binding protein 1, and interphotoreceptor retinoid-binding protein. The results support the hypothesis that a visual cycle protein complex may serve in the localization and release of 11-cis-retinoid in the apical RPE. PMID- 15336506 TI - Expression of glial fibrillary acidic protein in primary cultures of human Muller cells. AB - Glial fibrillary acidic protein (GFAP) is an intermediate filament protein which is primarily found in astrocytes. However, in eye diseases or when eye injuries occur, GFAP is expressed in large quantities in retinal Muller cells. The mechanism for this altered expression is unknown, but presumably involves injury dependent signaling. The purpose of this study was to investigate regulation of GFAP gene expression in human Muller cells in vitro. Immunofluorescence, western blot, RT-PCR and Northern blot analyses were used to demonstrate the expression of GFAP in cultured Muller cells. Plasmids bearing various segments of the human GFAP promoter fused to a CAT reporter gene were used to transfect primary cultures of human Muller cells as well as the non-glial cell lines 293T and HeLa. Transcription directed by the GFAP promoter was found to be more than 50-times stronger in the Muller cells than in either of the non-glial cell lines, consistent with the data for endogenous GFAP expression. To our knowledge, this is the first study demonstrating GFAP gene regulation in human Muller cells. By examining the transcriptional activity of various segments from the human GFAP promoter, it can be concluded that the GFAP gene is differently regulated in Muller cells compared to glioma cell lines from the central nervous system (CNS). PMID- 15336507 TI - Human, but not bovine, photoreceptor outer segments prime human retinal pigment epithelial cells for metabolic activation and massive oxidant release in response to lipopolysaccharide and interferon-gamma. AB - Reactive oxygen metabolites (ROMs) may contribute to several eye diseases, such as age-related macular degeneration, although the underlying mechanisms are unclear. The present study shows that human photoreceptor outer segments (POS) prime human retinal pigment epithelial (RPE) cells for massive ROM release in response to lipopolysaccharide (LPS) and interferon-gamma. However, no ROM priming of human RPE cells is observed for bovine POS. ROM production appears to be linked with underlying metabolic oscillations involving the hexose monophosphate shunt. PMID- 15336508 TI - TGFbeta2 influences alpha5-beta1 integrin distribution in human lens cells. AB - TGFbeta plays a central role in posterior capsule opacification, in which cell proliferation and matrix deposition, accompanied by capsular wrinkling, are largely responsible for the increased light scatter involved. Human FHL124 cells were plated onto uncoated glass coverslips to form circular patches so that the central cells reached confluency while the peripheral cells grew outwards. Cell patches were exposed to serum free (SF) EMEM (control) or TGFbeta supplemented (10 ng ml(-1)) EMEM. Fibronectin (Fn), alpha5beta1 integrin and F-actin were localized by immunofluorescence techniques and analysed by confocal microscopy. In the confluent, central cells in SF medium alpha5beta1 showed a punctate distribution while Fn was present in strongly staining fibres. TGFbeta had no effect on integrin or Fn distribution in confluent cells. In the peripheral, motile cells of the patches in SF conditions alpha5beta1 was localized in well defined focal adhesion plaques at the ends of actin stress fibres, while Fn was distributed in a punctate perinuclear pattern. TGFbeta had a profound dispersing effect on the integrin causing a widespread distribution of alpha5beta1 in the membrane with no apparent association with the actin filaments. The cells had a more fibroblastic morphology with increased deposition of Fn near the nucleus. All the TGFbeta-induced changes were inhibited by the TGFbeta antibody CAT152 (Cambridge Antibody Technology). Culture with a function-blocking alpha5 antibody or Fn antibody resulted in detachment of the peripheral cells from the patches, but the central cells remained intact. The patch culture method therefore provides a convenient means of investigating the differences between confluent and growing lens cells both in terms of the patterns of alpha5beta1 integrin and Fn and also in the response of the molecular arrangements of both to TGFbeta2. PMID- 15336509 TI - Comment on 'The photosensitiser xanthurenic acid is not present in normal human lenses' by P.G. Hains et al. [Exp. Eye Res. 77 (2003) 547-553]. PMID- 15336511 TI - Approaching a consensus cognitive battery for clinical trials in schizophrenia: the NIMH-MATRICS conference to select cognitive domains and test criteria. AB - To stimulate the development of new drugs for the cognitive deficits of schizophrenia, the National Institute of Mental Health (NIMH) established the Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS) initiative. This article presents an overview of decisions from the first MATRICS consensus conference. The goals of the meeting were to 1) identify the cognitive domains that should be represented in a consensus cognitive battery and 2) prioritize key criteria for selection of tests for the battery. Seven cognitive domains were selected based on a review of the literature and input from experts: working memory, attention/vigilance, verbal learning and memory, visual learning and memory, reasoning and problem solving, speed of processing, and social cognition. Based on discussions at this meeting, five criteria were considered essential for test selection: good test-retest reliability, high utility as a repeated measure, relationship to functional outcome, potential response to pharmacologic agents, and practicality/tolerability. The results from this meeting constitute the initial steps for reaching a consensus cognitive battery for clinical trials in schizophrenia. PMID- 15336512 TI - Animal modeling dual diagnosis schizophrenia: sensitization to cocaine in rats with neonatal ventral hippocampal lesions. AB - BACKGROUND: Increased substance disorder comorbidity in schizophrenia may reflect greater vulnerability to addictive processes because of inherent neurocircuit dysfunction in the schizophrenic brain. METHODS: To further explore this hypothesis, we used neonatal ventral hippocampal lesions (NVHL) as a rat model of schizophrenia and assessed locomotor sensitization to cocaine (15 mg/kg) in adulthood. RESULTS: The NVHL animals showed greater activity in response to an initial cocaine injection compared with sham and saline-treated groups. With daily cocaine injections over 7 days, NVHL rats showed elevated locomotor sensitization curves with greater fluctuations in the intersession changes in activity between days 4 and 7. In a single session 4 weeks later, NVHL compared with SHAM rats showed maintenance of cocaine-associated hyperactivity, as if superimposed on long-term sensitization effects present in both groups. CONCLUSIONS: In a neurodevelopmental model of schizophrenia, the locomotor effects of cocaine were augmented on initial and repeated doses, with emergence of irregularity in sensitization-related changes in activity in the short term and perseverance of augmented effects in the long term. Altered patterns of behavioral sensitization, as a possible correlate of greater addiction vulnerability, can occur as a by-product of neural systems dysfunction responsible for major psychiatric syndromes. PMID- 15336513 TI - Increased neurogenesis in a rat ketamine model of schizophrenia. AB - BACKGROUND: Growing evidence implicates abnormal neurodevelopment in schizophrenia, which manifests itself, for example, in reduced volume and cellular disarray of the hippocampus. This prompted us to investigate if there are indications of an altered neurodevelopment in this brain region. While neuron birth is largely completed by the end of gestation, granule neurons of the dentate gyrus are generated throughout life, thus offering an opportunity to investigate neurogenesis postnatally. METHODS: We investigated whether repeated application of subanesthetic doses of the noncompetitive N-methyl-D-aspartate receptor antagonist ketamine, which has been shown to mimic model aspects of schizophrenia in animals, affects the hippocampal neurogenesis detected by bromodeoxyuridine incorporation. Cells were identified by immunocytochemistry. RESULTS: Subanesthetic doses of ketamine applied subchronically enhance neurogenesis in the hippocampal subgranular zone. CONCLUSIONS: In our animal model of schizophrenia, ketamine may evoke its stimulating effect on neurogenesis via a block of the N-methyl-D-aspartate receptor directly by reducing the c-Fos/c Jun expression, resulting in a depression of the AP1 transcription factor complex and/or by a reduced nitric oxide production or an enhanced serotonergic activity. The newly formed neurons are not able to overcome the schizophrenia-related loss of parvalbumin expressing neurons and the behavioral abnormalities indicating that their functional integration is crucial. PMID- 15336514 TI - Abnormal activity patterns in premotor cortex during sequence learning in autistic patients. AB - BACKGROUND: Evidence for frontal abnormality in autism has accumulated in recent years. Our own studies have shown abnormal activation in prefrontal cortex during finger tapping and visuomotor coordination. Studies in healthy adults suggest reduced premotor and increased prefrontal activity during advanced learning stages. We examined hemodynamic changes during visuomotor learning in autistic patients. METHODS: We studied eight high-functioning autistic patients and eight control subjects during learning of an 8-digit sequence over a period of 8 min, using functional magnetic resonance imaging. RESULTS: Autistic patients showed overall less prefrontal activation during late visuomotor learning; however, the main finding was a complementary one of enhanced activation in right pericentral and premotor cortex. In the autism group, Brodmann areas 3, 4, and 6 of the right hemisphere became more involved during late learning stages (trials 25-48), compared with early stages (trials 1-24). This effect was not seen in the control group. CONCLUSIONS: Our findings suggests that in autistic patients 1) primary sensorimotor and premotor cortex, which is normally predominant in early stages of visuomotor learning, plays an atypical role in later stages, even when learning is evident; and 2) handedness and side of execution interact with asymmetry of visuomotor learning activations, contrary to what is seen in normal adults. PMID- 15336515 TI - Inhibitory deficits in ocular motor behavior in adults with attention deficit/hyperactivity disorder. AB - BACKGROUND: Many of the symptoms of attention-deficit/hyperactivity disorder (ADHD) have been attributed to deficits in behavioral inhibition mediated by the frontostriatal system. The ability to suppress unwanted saccadic eye movements is mediated by prefrontal cortex-basal ganglia circuitry and thus constitutes a useful measure of inhibitory ability. METHODS: To evaluate the functional integrity of this circuitry in ADHD, adult ADHD subjects unmedicated for at least 48 hours and normal comparison adults were studied by means of a comprehensive battery of ocular motor paradigms. RESULTS: On a prosaccade task, in which subjects were required to generate saccades toward a peripheral visual target after a short stimulus-free interval, ADHD subjects generated significantly more of anticipatory (premature) saccades (reaction time <90 msec) and of saccades toward the target on catch trials, in which they were supposed to inhibit eye movements. On the antisaccade task, in which they were required to inhibit gazing toward the target while moving their eyes in the opposite direction, ADHD subjects made significantly more directional errors than normal adults. The performance of ADHD adults was consistent with deficits in saccadic inhibition. CONCLUSIONS: Given the recent evidence for the interdependence between the brain systems mediating visual attention and ocular motor behavior, these findings support the notion that deficits in inhibitory mechanisms might underlie the inattention characteristic of ADHD. These results also implicate abnormalities in prefrontal cortex-basal ganglia circuitry in ADHD. PMID- 15336516 TI - Lithium and valproic acid treatment effects on brain chemistry in bipolar disorder. AB - BACKGROUND: Prior work reported elevated gray matter (GM) lactate and Glx (glutamate + glutamine + GABA) concentrations in unmedicated patients with bipolar disorder (BP) compared with healthy controls (HC). This study examined whether lithium (Li) and valproic acid (VPA) treatment modulated these chemicals. METHODS: A subset of previously reported BP patients were treated with Li (n = 12, 3.6 +/- 1.9 months) or VPA (n = 9, 1.4 +/- 1.7 months) and compared untreated HC subjects (n = 12, 2.9 +/- 2.4 months) using proton echo-planar spectroscopic imaging. Regression analyses (voxel gray/white composition by chemistry) were performed at each time point, and change scores computed. Metabolite relaxation and regions of interest (ROI) were also examined. RESULTS: Across treatment, Li treated BP subjects demonstrated GM Glx decreases (Li-HC, p =.08; Li-VPA p =.04) and GM myo-inositol increases (Li-HC p =.07; Li-VPA p =.12). Other measures were not significant. Serum Li levels were positively correlated with Glx decreases at the trend level. CONCLUSIONS: Li treatment of BP was associated with specific GM Glx decreases and myo-inositol increases. Findings are discussed in the context of cellular mechanisms postulated to underlie Li and VPA therapeutic efficacy. PMID- 15336517 TI - Neural mechanisms of working memory in ecstasy (MDMA) users who continue or discontinue ecstasy and amphetamine use: evidence from an 18-month longitudinal functional magnetic resonance imaging study. AB - BACKGROUND: Working memory processing in ecstasy (3,4 methylenedioxymethamphetamine) users is associated with neural alterations as measured by functional magnetic resonance imaging. Here, we examined whether cortical activation patterns change after prolonged periods of continued use or abstinence from ecstasy and amphetamine. METHODS: We used an n-back task and functional magnetic resonance imaging in 17 ecstasy users at baseline (t(1)) and after 18 months (t(2)). Based on the reported drug use at t(2) we separated subjects with continued ecstasy and amphetamine use from subjects reporting abstinence during the follow-up period (n = 9 and n = 8, respectively). RESULTS: At baseline both groups had similar task performance and similar cortical activation patterns. Task performance remained unchanged in both groups. Furthermore, there were no detectable functional magnetic resonance imaging signal changes from t(1) to t(2) in the follow-up abstinent group. However, the continuing users showed a dose-dependent increased parietal activation for the 2 back task after the follow-up period. CONCLUSIONS: Our data suggest that ecstasy use, particularly in high doses, is associated with greater parietal activation during working memory performance. An altered activation pattern might appear before changes in cognitive performance become apparent and, hence, may reflect an early stage of neuronal injury from the neurotoxic drug ecstasy. PMID- 15336518 TI - Smaller hippocampal volume in Dutch police officers with posttraumatic stress disorder. AB - BACKGROUND: Previous magnetic resonance imaging studies of posttraumatic stress disorder (PTSD) have reported smaller hippocampal volume, especially in war and sexual abuse victims. Our aim was to assess hippocampal volume in traumatized police officers with and without PTSD in the absence of alcohol abuse and moderate to severe major depression. METHODS: In a case-matched control study, 14 police officers with current PTSD and 14 traumatized police officers without lifetime PTSD were examined using magnetic resonance imaging. Three temporal lobe areas were manually segmented: hippocampus, amygdala, and parahippocampal gyrus. Volumetric analysis was used to measure gray matter, white matter, and cerebrospinal fluid. RESULTS: After controlling for total brain volume, the hippocampal volume in the PTSD group was significantly smaller in comparison with the traumatized control group (total 10.6%; left 12.6%). Volumes of amygdala, parahippocampal gyrus, gray matter, white matter, and cerebrospinal fluid were not significantly altered. A significant negative correlation was found between reexperiencing symptoms and hippocampal volume in the PTSD group. CONCLUSIONS: We confirmed previous findings of smaller hippocampal volume in PTSD in a new population made up of police officers, excluding comorbidity as a confounder. The finding of smaller hippocampal volume was specific to PTSD. PMID- 15336519 TI - The neural correlates of mate competition in dominant male rhesus macaques. AB - BACKGROUND: Male sexual jealousy provoked by threatened exclusive access to a female mate is a frequently reported motive in cases involving spousal abuse. Dominant male rhesus macaques also respond aggressively to threats to mating exclusivity. METHODS: Nine dominant male monkeys were injected with [(18)F] fluorodeoxyglucose ([(18)F]-FDG) and then exposed to one of two conditions: a "challenge" condition in which they witnessed a potential sexual interaction between their female consort and a rival male, and a control condition in which the consort was present without the rival male. After the brain uptake period for [(18)F]-FDG, dominant males were sedated, blood samples were drawn, and regional cerebral glucose metabolism was measured with positron emission tomographic imaging. RESULTS: Males that showed larger increases in plasma testosterone in the challenge condition showed more aggression and greater activation in the central gray matter of the midbrain, a brain area rich in androgen receptors. The challenge condition was associated with activation in both right superior temporal sulcus and right amygdala, which might relate to increased social vigilance and anxiety, respectively. CONCLUSIONS: Sexual jealousy in male humans is also often accompanied by vigilance behavior and anxiety and might recruit a similar neural network to that described here. PMID- 15336520 TI - No association between the Val66Met polymorphism of the brain-derived neurotrophic factor gene and bipolar disorder in a Japanese population: a multicenter study. AB - BACKGROUND: Two previous studies reported a significant association between a missense polymorphism (Val66Met) in the brain-derived neurotrophic factor (BDNF) gene and bipolar disorder; however, contradictory negative results have also been reported, necessitating further investigation. METHODS: We organized a multicenter study of a relatively large sample of 519 patients with bipolar disorder (according to DSM-IV criteria) and 588 control subjects matched for gender, age, and ethnicity (Japanese). Genotyping was done by polymerase chain reaction-based restriction fragment length polymorphism or direct sequencing. RESULTS: The genotype distributions and allele frequencies were similar among the patients and control subjects. Even if the possible relationships of the polymorphism with several clinical variables (i.e., bipolar I or II, presence of psychotic features, family history, and age of onset) were examined, no variable was related to the polymorphism. CONCLUSIONS: The Val66Met polymorphism of the BDNF gene is unrelated to the development or clinical features of bipolar disorder, at least in a Japanese population. PMID- 15336521 TI - Detecting DNA-binding of proteins in vivo by UV-crosslinking and immunoprecipitation. AB - The temporal and spatial binding of proteins on DNA is important to the regulation of genome expression and maintenance. However, examining how the protein-DNA complexes assemble in living cells is challenging. The development of UV-crosslinking/immunoprecipitation (UV-X-ChIP) technique and the progress of its applications show the powerful potential of this method in detecting such binding behavior in vivo. UV light is a zero length crosslinker and is believed to produce less perturbation of the complex than chemical crosslinker. The use of UV laser as UV light source allows the number of photons required for crosslinking to be delivered in nano- or pico- or femtosecond intervals, extremely shortening the irradiation time and achieving higher crosslinking efficiency than conventional UV lamp, thus being well suitable for kinetic studies. UV-X-ChIP technique has been successfully applied on the study of DNA replication, transcription, chromatin structure, and genome-wide location of DNA-binding proteins. PMID- 15336522 TI - Molecular identification and characterization of an alginate-binding protein on the cell surface of Sphingomonas sp. A1. AB - Cells of Sphingomonas sp. A1 (strain A1) directly incorporate a macromolecule, alginate, into cytoplasm through a biosystem, or "super-channel," consisting of a pit on the cell surface, alginate-binding proteins in periplasm, and an ABC transporter in the inner membrane. The pit functions as a concentrator for extracellular alginate. Through differential display analysis, a protein (p8) with a molecular mass of 20kDa and a pI of 7.4 was found to be inducibly expressed in the outer membrane of alginate-grown cells. The gene coding for p8 was identified in the genome of strain A1 and shown to be similar to that for the polyhydroxyalkanoate granule-associated protein of Ralstonia eutropha. The disruptant of p8 gene showed significant growth retardation in the alginate medium. An overexpression system for p8 was constructed in Escherichia coli, and the protein was purified and characterized. Surface plasmon resonance biosensor analysis indicated that p8 is able to bind alginate most efficiently at pH 4.0. The above results indicate that p8 is a cell surface protein able to bind alginate and facilitates the concentration of alginate in the pit on the cell surface of strain A1. PMID- 15336523 TI - Endotoxin unmasks the role of gap junctions in the liver. AB - Gap junctions are thought to be necessary for proper tissue function. However, no clear hepatic phenotype has been described in patients lacking connexin 32 (Cx32), the principal gap junction in liver. To determine the physiological role of Cx32 in liver, we compared the response of wild type and Cx32-deficient mice to endotoxin, since this stress increases serum levels of hormones that bind to receptors that are asymmetrically distributed across the hepatic lobule. In hepatocyte couplets isolated from wild type mice, most hepatocytes could transfer microinjected dye to their neighbor even after treatment with endotoxin. Dye transfer was not observed in Cx32-deficient couplets. Treatment of hepatocyte couplets from wild type mice with vasopressin induced calcium (Ca(2+)) waves that crossed the couplets in a concentration-dependent fashion, but the delay in transmission was markedly prolonged at all concentrations in Cx32-deficient couplets. Expression of the vasopressin receptor and the inositol 1,4,5 trisphosphate receptor was not decreased by endotoxin or in Cx32-deficient couplets. Finally, endotoxin caused transient hypoglycemia and cholestasis in wild type animals, but hypoglycemia was slightly prolonged and cholestasis was much worse in Cx32-deficient mice treated with endotoxin. The hepatic response to endotoxin is markedly impaired in the absence of Cx32. Thus, an important role of gap junctions in the liver is to assure integrated and uniform tissue response in times of stress. PMID- 15336524 TI - Activation of NF-kappaB is involved in 6-hydroxydopamine-but not MPP+ -induced dopaminergic neuronal cell death: its potential role as a survival determinant. AB - The nuclear factor-kappaB (NF-kappaB) family plays an important role in the control of the apoptotic response. Its activation has been demonstrated in both neurons and glial cells in many neurological disorders. In the present study, we specifically examined whether and to what extent NF-kappaB activation is involved in culture models of Parkinson's disease following exposure of MN9D dopaminergic neuronal cells to 6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenyl-4 phenylpyridinium ion (MPP(+)). Both analysis by immunocytochemistry and of immunoblots revealed that NF-kappaB-p65 was translocated into the nuclei following 6-OHDA but not MPP(+)-treatment. A time-dependent activation of NF kappaB induced by 6-OHDA but not MPP(+) was also demonstrated by an electrophoretic mobility shift assay. A competition assay indicated that not only NF-kappaB-p65 but also -p50 is involved in 6-OHDA-induced NF-kappaB activity. Co treatment with an antioxidant, N-acetyl-l-cysteine, blocked 6-OHDA-induced activation of NF-kappaB signaling. In the presence of an NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), 6-OHDA-induced cell death was accelerated while PDTC did not affect MPP(+)-induced cell death. Our data may point to a drug specific activation of NF-kappaB as a survival determinant for dopaminergic neurons. PMID- 15336525 TI - Characterization of the Arabidopsis thaliana Arath;CDC25 dual-specificity tyrosine phosphatase. AB - CDC25 enzymes are dual-specificity phosphatases involved in the regulation of the cell cycle. No CDC25 enzymes have been described in higher plant organisms. We report here the characterization of an Arabidopsis thaliana CDC25 enzyme, constituted by a sole catalytic domain and devoid of the N-terminal regulatory region found in the human CDC25. We describe the recombinant expression in Escherichia coli of the Arath;CDC25 and its purification for activity assay and structure determination by NMR. The recombinant enzyme has a tyrosine phosphatase activity towards an artificial substrate, a NMR characterization equally concludes to its correct folding. The secondary structure of the protein was predicted on the basis of the assigned chemical shift of (1)H, (15)N, and (13)C backbone atoms of the protein. The presence of a metal ion in the C-terminus of this new protein points to a zinc finger, and sequence homology indicates that this new structural element might be conserved in related plant homologs. PMID- 15336526 TI - Water channel aquaporin-2 directly binds to actin. AB - Water channel aquaporin-2 (AQP2) strictly regulates body water homeostasis in mammals. Trafficking of AQP2 to the apical membrane is critical to the reabsorption of water in renal collecting ducts. Controlled apical positioning of AQP2 suggests the interaction of AQP2 with other proteins. To isolate AQP2 binding proteins, immunoaffinity chromatography of extracts from rat kidney papilla was performed using a column covalently coupled with anti-AQP2 antibody. Using this method 42-kDa protein was purified and subsequently identified as beta and gamma-isoforms of actin by two-dimensional gel analysis and matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. AQP2 was indeed coimmunoprecipitated with actin from cell lysates of rat kidney papilla. In addition, surface plasmon resonance analyses showed that the C-terminal fragment of AQP2 strongly bound to actin and the K(D) value was 3.18x10(-8)M. In this experiment we have elucidated the direct binding of channel protein AQP2 to cytoskeletal protein actin, providing a novel mechanism for trafficking of not only AQP2 but also recycling channel proteins. PMID- 15336527 TI - Permeability of lipid membranes to dioxygen. AB - It is commonly supposed that dioxygen (O(2)) transport through biomembranes is ensured by the high permeability of a lipid bilayer in which O(2) diffusion mobility is close to that in water. However, the fact that microviscosity of lipid membranes is higher than that of water by two to three orders of magnitude speaks against this concept. Therefore, in this work we investigated the influence of surface lipid monolayers on oxygen diffusion flow directed from air to aqueous phase. We show that for lipid monolayers, the O(2) permeability coefficients are within the range of 10(-4) to 10(-5)m/s. These values are three to four orders of magnitude lower than has been previously thought, indicating that lipid membranes constitute a considerable barrier to O(2) diffusion. From this, we suggest that membranes of aerobic organisms contain O(2) channels to ensure the high-volume transmembrane O(2) flows. PMID- 15336528 TI - Action of solamargine on TNFs and cisplatin-resistant human lung cancer cells. AB - A loss of TNF receptors expression has been found in advanced lung cancers, and human A549 lung adenocarcinoma cells are resistant to the cytotoxic effects of TNF-alpha and cisplatin. Here, the mechanisms of the drug resistance of A549 were extensively studied by gene modulation of the cells by solamargine (SM) which was isolated from Solanum incanum herb. SM induced morphological changes of chromatin condensation, DNA fragmentation, and sub-G(1) peak in a DNA histogram of A549 cells, indicating cell death by apoptosis. SM elevated the expressions of TNF-R1 and -R2 and overcame the resistance of A549 cells to TNF-alpha and -beta. The recruitment of TRADD, FADD, and activation of caspase-8 and -3 in SM-treated A549 cells evidenced the activation of TNFRs signal transduction. In addition, release of cytochrome c from mitochondria, down-expression of Bcl-2 and Bcl-x(L), up regulation of Bax, and caspase-9 activities were observed in SM-treated A549 cells. Combinational treatment of SM and cisplatin synergistically enhanced caspase-8, -9, and -3 activities in A549 cells. Thus, SM sensitizes A549 cells through TNFRs and mitochondria-mediated pathways and may have anticancer potential against TNFs- and cisplatin-resistance lung cancer cells. PMID- 15336529 TI - Expression of metalloproteinases and inhibitors in the differentiation of P19CL6 cells into cardiac myocytes. AB - P19CL6 are a clonal derivative of P19 embryonal carcinoma cells, a euploid, multipotent mouse cell line, that differentiate efficiently into cardiac myocytes, with spontaneous beating evident within 10 days, following DMSO treatment. Using real-time quantitative RT-PCR we have profiled the expression of the complete matrix metalloproteinase and tissue inhibitor of metalloproteinase gene families during P19CL6 differentiation to cardiac myocytes. The genes subdivide into eight groups based upon their expression profile. Their expression was both qualitatively and quantitatively highly homologous to that seen during mouse heart development. PMID- 15336530 TI - The solubility and stability of recombinant proteins are increased by their fusion to NusA. AB - The new bacterial vector pETM60 enables the expression of His-tagged recombinant proteins fused to the C-terminus of NusA through a TEV protease recognition sequence. Three sequences coding for two protein domains (Xklp3A and Tep3Ag) and one membrane-bound viral protein (E8R) could not be expressed in a soluble form in bacteria. Their GST-fusions were mostly soluble but quickly degraded during purification. The same sequences cloned in pETM60 were efficiently purified by metal affinity and recovered soluble after the removal of the fusion partner. The NusA-fused constructs enabled to yield 13-20mg of fusion protein per litre of culture and 2.5-5mg of pure protein per litre of culture. Structural analysis indicated that the purified proteins were monodispersed and correctly folded. NusA has been used to raise antibodies that have been successfully used for Western blot and immunoprecipitation of NusA fusion proteins. PMID- 15336531 TI - Treatment with leucine stimulates the production of hepatocyte growth factor in vivo. AB - Hepatocyte growth factor (HGF) has pleiotropic effects. Up-regulation of HGF activity in vivo may be beneficial. Branched-chain amino acids (BCAAs) are known to modulate various cellular functions. When starved rats received intraperitoneal injections of valine, leucine or isoleucine, only leucine treatment increased both hepatic and circulating levels of HGF in a dose dependent manner, up to 1.5 and 2.3 times higher, respectively, than in controls. When young growing rats with free access to food were injected with leucine once a day for a week, HGF levels and liver weights were significantly higher than those of control rats. Furthermore, 1 week of leucine treatment of adult rats resulted in elevated serum albumin levels with an increase in HGF levels. Taken together with our previous report showing that leucine stimulates HGF production by hepatic stellate cells in culture, leucine, among BCAAs, may induce an increase in HGF production by the liver in vivo. PMID- 15336532 TI - Celastrol inhibits pro-inflammatory cytokine secretion in Crohn's disease biopsies. AB - Crohn's disease is a chronic intestinal inflammatory process. In modern therapy, TNF-alpha inhibition is the main goal. The aim here is to characterize the effects of Celastrol, a pentacyclic-triterpene, on the secretion of inflammatory cytokines by LPS-activated human cells. Celastrol dose-dependently inhibited the secretion of all tested pro-inflammatory cytokines with IC(50) in the nanomolar range. Effect not related to glucocorticoid receptor activity is shown by competition experiments with the steroid antagonist RU486. Celastrol inhibited the pro-inflammatory cytokine secretion from mucosal inflammatory biopsies from Crohn's disease patients. Cytometry emphasized that for all tested pro inflammatory cytokines, CD33(+) cells are the most sensitive. Quantitative-PCR and confocal analysis on a human monocytic cell line indicated that Celastrol acts at the transcriptional level by inhibiting LPS-induced NF-kappaB translocation. Celastrol might be a putative anti-inflammatory drug in the treatment of inflammatory diseases, given its inhibition of cytokine production by intestinal biopsies from Crohn's disease patients. PMID- 15336533 TI - A genome-wide and nonredundant mouse transcription factor database. AB - Here we describe the development of a genome-wide and nonredundant mouse transcription factor database and its viewer (http://genome.gsc.riken.gp/TFdb/). We systematically selected transcription factors with DNA-binding properties and their regulators on the basis of their LocusLink and Gene Ontology annotations. We also incorporated into our database information regarding the corresponding available cDNA clones and their structural properties. Because of these features, our database is unique and may provide useful information for systematic genome wide studies of transcriptional regulation. PMID- 15336534 TI - Is mRNA and protein level of CD46 altered in measles virus vaccine strain S191 infected cells? AB - Previous research showed that the expression of measles virus receptor CD46 was downregulated after expression of measles virus hemagglutinin protein on the surface of the virus infected cell or triggered by infected cell-to-cell contact. We reported here that the mRNA level of CD46 in MV infected cells was not changed which was tested by real-time quantitative PCR. To further analyse the surface expression alteration of CD46 after MV infection, flow cytometric analysis and indirect immunofluorescence were used to detect the protein level of CD46. Altogether, our results provided a demonstration that the expression of CD46 was not downregulated by the infection of MV strain S191 both on mRNA level and cellular surface protein level. Previous results reported that the "downregulation" of CD46 expression on the cell surface may take place because H protein masks the antibody recognition site on CD46 which results in "downregulation" of the expression of CD46. PMID- 15336535 TI - The 7472insC mtDNA mutation impairs 5' and 3' processing of tRNA(Ser(UCN)). AB - The deafness-associated 7472insC mtDNA mutation was previously shown to decrease the steady-state level of tRNA(Ser(UCN)) post-transcriptionally. To identify the affected tRNA maturation step(s) we analysed the effects of the mutation on processing in vivo and in vitro. tRNA(Ser(UCN)) from cybrid cells homoplasmic for 7472insC contained a high frequency (>11%) of molecules misprocessed at one or both termini. In vitro assays using partially purified HeLa cell RNase P and mitochondrial tRNA 3' processing endonuclease (tRNase Z) confirmed that the efficiency of both 5' and 3' processing was impaired. A mutant precursor not already processed at the 5' end was poorly processed in vitro by tRNase Z. Misprocessing at the 3' end further impaired the efficiency and accuracy of 5' processing of the mutant substrate. The mutation thus appears to affect several distinct, but interdependent, RNA processing steps, with the predicted outcome dependent on the exact processing pathway operating in vivo. PMID- 15336536 TI - Baculovirus-mediated expression and isolation of human ribosomal phosphoprotein P0 carrying a GST-tag in a functional state. AB - We constructed an overexpression system for human ribosomal phosphoprotein P0, together with P1 and P2, which is crucially important for translation. Genes for these proteins, fused with the glutathione S-transferase (GST)-tag at the N terminus, were inserted into baculovirus and introduced to insect cells. The fusion proteins, but not the proteins without the tag, were efficiently expressed into cells as soluble forms. The fusion protein GST.P0 as well as GST.P1/GST.P2 was phosphorylated in cells as detected by incorporation of (32)P and reactivity with monoclonal anti-phosphoserine antibody. GST.P0 expressed in insect cells, but not the protein obtained in Escherichia coli, had the ability to form a complex with P1 and P2 proteins and to bind to 28S rRNA. Moreover, the GST.P0-P1 P2 complex participated in high eEF-2-dependent GTPase activity. Baculovirus expression systems appear to provide recombinant human P0 samples that can be used for studies on the structure and function. PMID- 15336537 TI - Determinants of apical loop-internal loop RNA-RNA interactions involving the HCV IRES. AB - Domain II of the hepatitis C virus internal ribosome entry site is a major RNA structure involved in the viral mRNA translation. It comprises four different structural domains. We performed in vitro selection against the apical loop of the domain II and we identified RNA aptamers folding as an imperfect hairpin with an internal loop of interacting with the apical loop of the domain II. This RNA RNA interaction creates apical loop-internal loop complex. The aptamer binds the target with an apparent K(d) of 35nM. In this study, the main structural elements of the target and the aptamer involved in the formation of the complex are characterized by mutation, deletion, and RNase probing analysis. We demonstrate that a complementary loop flanked by G,C rich upper and lower stems are crucial for such RNA-RNA interactions. PMID- 15336538 TI - cDNA microarray analysis of early gene expression profiles associated with hepatitis B virus X protein-mediated hepatocarcinogenesis. AB - Chronic hepatitis B virus (HBV) infection is one of the major causes of hepatocellular carcinoma. HBV encodes an oncogenic hepatitis B virus X protein (HBx), which can transactivate host cell transcriptional machinery and mediate cellular transformation. To disclose the early genetic response in HBx-mediated transformation process, we constructed a conditional HBx-expressing hepatocyte cell line, which allows us to compare the gene expression profiles under controllable HBx induction. A cDNA microarray containing more than 8700 mouse genes and ESTs was utilized to examine the gene expression profiles. We identified 260 candidate genes and 259 ESTs which have shown aberrant expression under HBx induction. Most of them are involved in signal transduction pathway, cell cycle control, metastasis, transcriptional regulation, immune response, and metabolism. These results provide additional insight into early cellular targets of HBx, which could give us a better understanding of the function of HBx and their progressive changes during HBx-mediated hepatocarcinogenesis. PMID- 15336539 TI - Drug uptake and pharmacological modulation of drug sensitivity in leukemia by AQP9. AB - Leukemia is the most common childhood cancer. Trisenox, the active ingredient of which is trivalent arsenic, is the first line of treatment for acute promyelocytic leukemia. Since drug action usually requires uptake of the drug, it is of importance to determine the transport system responsible for Trisenox uptake. Recently, human aquaglyceroporin 9 (AQP9) has been shown to transport As(III) in Xenopus oocytes. In this study we report to show that AQP9 expression modulates the drug sensitivity of leukemic cells. AQP9 was transfected into the chronic myelogenous leukemia cell line K562. The transfectants became hypersensitive to Trisenox and Sb(III). The promyelocytic leukemia cell line HL60 treated with vitamin D showed higher expression of AQP9 and hypersensitivity to Trisenox and Sb(III). This sensitivity was due to higher rates of uptake of the trivalent metalloids by the cell lines overexpressing AQP9. Trisenox hypersensitivity results from increased expression of AQP9 drug uptake system. The possibility of using pharmacological agents to increase expression of AQP9 gene delivers the promise of new therapies for the treatment of leukemia. Thus, drug hypersensitivity can be correlated with increased expression of the drug uptake system. This is the first demonstration that AQP9 can modulate drug sensitivity in cancer. PMID- 15336540 TI - Hepatic stellate cells lack AP-1 responsiveness to electrophiles and phorbol 12 myristate-13-acetate. AB - Stellate cell profibrotic gene induction and transdifferentiation are central events in liver fibrosis. Oxidative stress has been implicated as an activator of the transcription factors Nrf2 and AP-1 through shared kinase signaling pathways that also purportedly contribute to stellate cell activation. The present study examined the role of oxidative stress in ARE- and TRE-regulated gene induction in isolated hepatic stellate cells. Using a portion of the human Nqo1 promoter consisting of an ARE imbedded TRE, it was demonstrated that while the ARE was responsible for mediating inducible gene expression in response to the electrophiles 4-HNE and tBHQ, the TRE was refractory to induction by either electrophiles or PMA. It was demonstrated that stellate cells possess nuclear TRE binding proteins that were identified as JunB, JunD, Fra1, and Fra2, which were unaffected by either electrophiles or PMA treatment. This report demonstrates that, in contrast to the ARE, the TRE and its binding cognate AP-1 did not mediate independent gene induction in hepatic stellate cells. This observation is significant given the presumed importance attributed to AP-1 in mediating profibrogenic gene expression. PMID- 15336541 TI - Targeting of Bacillus anthracis interaction factors for human macrophages using two-dimensional gel electrophoresis. AB - Bacillus anthracis, a gram-positive, endospore-forming, aerobic rod-shaped bacterium, interacts with macrophages at various stages of the disease. Spore germination and the outgrowth of vegetative bacilli are crucial steps enabling the bacteria to proliferate actively and to synthesize the virulence factors leading to a massive septicemia. In this study, we performed a proteomic analysis and MALDI-TOF/MS were carried out to identify proteins using human macrophages infected with the spores of B. anthracis live-Sterne or inactivated-Sterne. We identified 21 proteins which are related to the infection of B. anthracis spores on human macrophages at the early stage events. These proteins function in processes such as cytoskeleton regulation, apoptosis, cell division, and protein degradation. Proteins such as PAK 2 revealed a relationship to apoptosis in human macrophages. These proteins play an important role in the macrophage survival and death on human macrophages with infected B. anthracis spores. PMID- 15336542 TI - Analysis of the type IV secretion system-dependent cell motility of Helicobacter pylori-infected epithelial cells. AB - The pathogenesis of Helicobacter pylori-associated disorders is strongly dependent on a specialized type IV secretion system (T4SS) encoded by the cag pathogenicity island (PAI). Cytotoxin-associated gene A (CagA) is the only known H. pylori protein translocated into the host cell followed by tyrosine phosphorylation through host protein kinases. H. pylori induces cellular processes which are either PAI- or CagA-dependent (e.g., cell motility), PAI dependent, but CagA-independent (e.g., interleukin-8 release), or PAI- and CagA independent (e.g., cyclooxygenase-2 release). Here, we investigated H. pylori strains mutated in single PAI genes of the wild type strain Hp26695 and their effects on cell motility. We found 17 gene products out of 27 PAI genes playing a superordinated role and five PAI-encoded proteins exhibiting a clearly critical role in motogenic host cell responses, whereas the remaining five PAI gene products had no significant influence on the motogenic response in reaction to H. pylori infection. This study clearly demonstrated that H. pylori-induced cell motility and invasive growth involve type IV secretion system-dependent signalling as well as translocated and phosphorylated CagA. These findings reveal a deeper insight in to the meaning of the T4SS of H. pylori for host cell motility. PMID- 15336543 TI - Characterization of the synthetic compatible solute homoectoine as a potent PCR enhancer. AB - Different substances such as dimethyl sulfoxide, tetramethylene sulfoxide, 2 pyrollidone, and the naturally occurring compatible solute betaine enhance PCR amplification of GC-rich DNA templates with high melting temperatures. In particular, cyclic compatible solutes outperform traditional PCR enhancers. We therefore investigated the effects that cyclic naturally occurring ectoine-type compatible solutes and their synthetic derivatives have on melting temperature of double-stranded DNA (dsDNA) and on PCR amplification of different templates. L ectoine, betaine, and derivatives of L-ectoine decreased, whereas beta hydroxyectoine increased, the melting temperature of dsDNA. The ability to decrease the melting temperature was greatest for homoectoine, a new synthetic derivative of l-ectoine. Furthermore, compatible solutes, especially homoectoine, enhanced PCR amplification of GC-rich DNA (72.6% GC content; effective range: 0.1 0.5M). PMID- 15336544 TI - Nickel, lead, and cadmium induce differential cellular responses in sea urchin embryos by activating the synthesis of different HSP70s. AB - Treatment with heavy metals, such as nickel, lead or cadmium, elicits different cellular stress responses according to the metal used and the length of treatment. In Paracentrotus lividus embryos the inducible forms of HSP70 (HSP70/72) are different in molecular mass from the constitutively expressed HSP75, and they can be used as markers of cellular stress. Even a short treatment with each metal induces the synthesis of HSP70/72 which remain stable for at least 20h and differ little in their isoelectric points. Continuous treatment from fertilization with nickel or lead produces late irregular pluteus embryos, with peak HSP70/72 synthesis at blastula followed by the arrest of synthesis by pluteus. On the contrary, the same treatment with cadmium induces continuous HSP70/72 synthesis and produces irregular gastrula embryos which then degenerate. Moreover, a long treatment induces over control embryos a slight increase in the amount of constitutive HSP75 during development while lead treatment depresses constitutive HSP75 at early stages and doubles its quantity at late stages. PMID- 15336545 TI - Adrenomedullin inhibits prostate cancer cell proliferation through a cAMP independent autocrine mechanism. AB - Adrenomedullin (AM) is a multifunctional peptide expressed in the normal and malignant prostate, and in prostate cancer cells. To elucidate the potential role of AM in prostate cancer, we have transfected the human AM gene into PC-3, DU 145, and LNCaP prostate cancer cells. Northern blot, Western blot, and radioimmunoassay techniques confirmed an increase in the synthesis and secretion of the 6kDa mature peptide, in the AM-transfected clones. Proliferation and cell cycle assays demonstrated that AM overexpression inhibited cell proliferation in PC-3 and LNCaP cells through a G0/G1 cell cycle arrest, but not in DU 145 cells. In vivo growth assays also confirmed that, at least in PC-3, AM produced a very significant reduction of tumor volume. In addition, the three cell lines expressed the CL/RCP/RAMP-2 receptor complex by RT-PCR, which suggests that AM peptide acts through an autocrine loop in prostate cancer cells. Although cAMP elevation is the most common pathway involved in AM signalling, stimulation of PC 3, DU 145, and LNCaP with synthetic AM did not increase intracellular cAMP. However, short-term stimulation of PC-3 cells with synthetic AM increased ERK1/2 activation. On the contrary, long-term stimulation, or AM overexpression, caused a reduction in the basal activation of ERK1/2. In summary, our results demonstrate that AM (either overexpressed or exogenously added) causes an inhibition of prostate cancer cell growth. This inhibition does not depend on changes in intracellular cAMP levels, but may be related to ERK1/2 activation. PMID- 15336546 TI - Fgf18 is required for embryonic lung alveolar development. AB - Fgf18 is abundantly expressed in mouse embryonic lungs. To elucidate the roles of Fgf18 in mouse embryonic lung development, we examined the Fgf18-/- embryonic lungs. Although the sizes of the Fgf18-/- lungs were a little smaller in appearance than those of wild-type lungs, neither proximal nor distal airway branching in the Fgf18-/- lungs was impaired. However, the Fgf18-/- lungs at E18.5 had reduced alveolar space, thicker interstitial mesenchymal compartments, and many embedded capillaries. Cell proliferation in the Fgf18-/- lungs was also transiently reduced around E17.5, although the expression of marker genes for lung epithelial cells in the Fgf18-/- lungs was not impaired. The present findings indicate that the Fgf18 plays roles in lung alveolar development during late embryonic lung development stages. The cell proliferation during the terminal saccular stage stimulated by Fgf18 might play roles in the remodeling of the distal lung. PMID- 15336547 TI - A role for hippocampal Rho-ROCK pathway in long-term spatial memory. AB - Morphological changes, including changes in size, shape, and number of synapses, in neurons have been observed in many species and are thought to be critical for long-term memory storage. Actin filaments are intimately involved in neuronal morphology and regulation of their dynamics can influence memory. Rho GTPase plays a prominent role in this process and has been implicated in both pre- and post-synaptic morphological changes. Therefore, we examined the effect of hippocampal manipulation of Rho and ROCK activity on performance in a spatial memory task. Post-training intrahippocampal infusion of an inhibitor of the downstream effector kinase p160ROCK impaired long-term memory. Furthermore, post training activation of Rho using lysophosphatidic acid (LPA) enhanced long-term spatial memory. This memory enhancing effect of LPA was not mediated via the Erk cascade, as no change in Erk phosphorylation was observed as a result of its administration. Our results demonstrate a role for the Rho-ROCK pathway in hippocampus-dependent spatial memory. PMID- 15336548 TI - Toxoplasma gondii-derived heat shock protein 70 stimulates the maturation of human monocyte-derived dendritic cells. AB - We investigated the role of Toxoplasma gondii-derived heat shock protein 70 (TgHSP70) as a dendritic cell (DC) maturation-inducing molecule. TgHSP70 induced the maturation of human monocyte-derived dendritic cells as determined by increased levels of surface markers, namely, CD40, CD80, CD86, and HLA-DR. Moreover, TgHSP70 also reduced phagocytic activity and increased the allostimulatory capacity of DCs, suggesting the functional maturation of DCs by TgHSP70. Maturation of DCs by TgHSP70 also elicited a significant increase in IL 12 production in a polymyxin B-insensitive manner. TgHSP70 also stimulated extracellular signal-regulated kinase and p38 kinase pathways in DCs, and TgHSP70 induced IL-12 production was inhibited by SB203580 but not by PD98059, thus indicating the role of p38 kinase in the maturation of DCs by TgHSP70. This study demonstrates the role of TgHSP70 in the functional maturation of DCs and suggests TgHSP70 as a useful molecule for the development of a vaccine against T. gondii. PMID- 15336549 TI - Catalytic mechanism of type 2 isopentenyl diphosphate:dimethylallyl diphosphate isomerase: verification of a redox role of the flavin cofactor in a reaction with no net redox change. AB - Type 2 isopentenyl diphosphate:dimethylallyl diphosphate isomerase requires redox co-enzymes, i.e., flavin mononucleotide (FMN) and NAD(P)H, for activity, although it catalyzes a non-redox reaction. Spectrometric studies and enzyme assays under anaerobic conditions indicate that FMN is reduced through the reaction and is sufficient for activity. The sole function of NAD(P)H appears to be the reduction of FMN since it could be replaced by an alternate reducing agent. When the enzyme was reconstructed with a flavin analogue, no activity was detected, suggesting that the isomerase reaction proceeds via a radical transfer mechanism. PMID- 15336550 TI - Microarray evaluation of EP4 receptor-mediated prostaglandin E2 suppression of 3T3-L1 adipocyte differentiation. AB - Prostaglandin E(2) (PGE(2)) has been shown to negatively regulate adipogenesis. To explore to what extent PGE(2) inhibits the differentiation of cells to adipocytes and to examine whether its effect could be due to EP4 receptor signaling, we used microarrays to analyze the gene expression profiles of 3T3-L1 cells exposed to a differentiation cocktail supplemented with PGE(2), AE1-329 (an EP4 agonist), or vehicle. The differentiation-associated responses in genes such as adipocytokines and enzymes related to lipid metabolism were largely weakened upon PGE(2) treatment. In particular, the expression of peroxisome proliferator activated receptor-gamma and CCAAT/enhancer binding protein-alpha, genes playing a central role in adipogenesis, was greatly suppressed. PGE(2) appears to be ineffective to a subclass of insulin target genes such as hexokinase 2 and phosphofructokinase. Similar responses were produced in the differentiation associated genes upon AE1-329 treatment. These results suggest that PGE(2) inhibits a crucial step of the adipocyte differentiation process by acting on the EP4 receptor in 3T3-L1 cells. PMID- 15336551 TI - Neurospheres induced from bone marrow stromal cells are multipotent for differentiation into neuron, astrocyte, and oligodendrocyte phenotypes. AB - Bone marrow stromal cells (MSCs) can be expanded rapidly in vitro and have the potential to be differentiated into neuronal, glial and endodermal cell types. However, induction for differentiation does not always have stable result. We present a new method for efficient induction and acquisition of neural progenitors, neuronal- and glial-like cells from MSCs. We demonstrate that rat MSCs can be induced to neurospheres and most cells are positive for nestin, which is an early marker of neuronal progenitors. In addition, we had success in proliferation of these neurospheres with undifferentiated characteristics and finally we could obtain large numbers of neuronal and glial phenotypes. Many of the cells expressed beta-tubulin III when they were cultivated with our method. MSCs can become a valuable cell source as an autograft for clinical application involving regeneration of the central nervous system. PMID- 15336552 TI - Inhibition of mitochondrial respiration by the anticancer agent 2 methoxyestradiol. AB - 2-Methoxyestradiol (2ME2), a naturally occurring metabolite of estradiol, is known to have antiproliferative, antiangiogenic, and proapoptotic activity. Mechanistically, 2ME2 has been shown to downregulate hypoxia-inducible factor 1alpha (HIF1alpha) and to induce apoptosis in tumour cells by generating reactive oxygen species (ROS). In this study we report that 2ME2 inhibits mitochondrial respiration in both intact cells and submitochondrial particles, and that this effect is due to inhibition of complex I of the mitochondrial electron transport chain (ETC). The prevention by 2ME2 of hypoxia-induced stabilisation of HIF1alpha in HEK293 cells was found not to be due to an effect on HIF1alpha synthesis but rather to an effect on protein degradation. This is in agreement with our recent observation using other inhibitors of mitochondrial respiration which bring about rapid degradation of HIF1alpha in hypoxia due to increased availability of oxygen and reactivation of prolyl hydroxylases. The concentrations of 2ME2 that inhibited complex I also induced the generation of ROS. 2ME2 did not, however, cause generation of ROS in 143B rho(-) cells, which lack a functional mitochondrial ETC. We conclude that inhibition of mitochondrial respiration explains, at least in part, the effect of 2ME2 on hypoxia-dependent HIF1alpha stabilisation and cellular ROS production. Since these actions of 2ME2 occur at higher concentrations than those known to inhibit cell proliferation, it remains to be established whether they contribute to its therapeutic effect. PMID- 15336553 TI - Restoration of protein synthesis in pancreatic cancer cells by trans-splicing ribozymes. AB - This report describes the use of trans-splicing ribozymes to restore p16 protein synthesis in pancreatic cancer cells. A group I intron ribozyme was designed to trans-splice the 2 base-deleted p16 transcripts with the wild-type sequence in a pancreatic cancer cell line, which originally produced no p16. Following transfection of the ribozyme construct in AsPC-1 cells, mutant p16 mRNA molecules were repaired and p16 protein synthesis restored. Moreover, these cells exhibited a reduced ability to grow, compared to the untransfected cells. The technology of ribozymes offers an advantage over gene replacement therapy because it maintains the cellular regulation of gene expression. These results indicate that group I intron ribozymes might prove useful towards the therapy of pancreatic cancer and in conjunction with the advancement of powerful delivery systems this technology will play a major role in the therapy of many diseases. PMID- 15336554 TI - Tenascin-C deposition requires beta3 integrin and Src. AB - In this study we now show that deposition of the mesenchymal matrix marker, tenascin-C (TN-C), is mediated through beta3 expression and activation of Src. There was a striking upregulation of TN-C matrix organization in cell lines expressing beta3 and activated Src when compared to cell lines with neither of these attributes. When beta3 function was suppressed so was the deposition of TN C. The same was true for function and activation of Src. When Src was inactive, the deposition of TN-C was low. We also determined that one of the downstream effectors of Src, MAPK, was also required to promote TN-C deposition. When MAPK activation was inhibited, TN-C deposition was also decreased. MMP activation is also implicated in TN-C deposition. The broad spectrum MMP inhibitor, GM6001, suppressed TN-C organization. These results indicate that beta3 integrin ligand binding and the activation of the Src/MAPK/MMP pathway modulate deposition of TN C. PMID- 15336555 TI - Pancreatic tumor cells influence the composition of the extracellular matrix. AB - The malignant behavior of cancers depends on the microenvironmental context. We investigated compositional alterations of the extracellular matrix (ECM) in pancreatic cancer, with special emphasis on the proteoglycans decorin, lumican, and versican. Compared with normal controls (n=18), marked overexpression of these proteoglycans was observed in pancreatic cancer tissues (n=30) by quantitative RT-PCR (p<0.0001). Immunohistochemistry revealed abundance of proteoglycans in the ECM of pancreatic cancer specimens, whereas tumor cells themselves were devoid of either decorin, lumican or versican. RT-PCR confirmed pancreatic stellate cells (PSCs) as the major source of these proteins. Interestingly, TGFbeta1 and conditioned medium derived from pancreatic cancer cell lines synergistically suppressed the expression of known anti-tumor factors decorin and lumican, but stimulated the expression of pro-metastatic factor versican in cultured PSCs. These findings indicate that malignant cells can actively influence the composition of the ECM through TGFbeta1 and other soluble factors, altering their microenvironment in a tumor-favorable way. PMID- 15336556 TI - Activation of alpha(M)beta(2)-mediated phagocytosis by HF3, a P-III class metalloproteinase isolated from the venom of Bothrops jararaca. AB - The integrin alpha(M)beta(2) regulates important cell functions in inflammation being the primary phagocytic receptor on macrophages. HF3, a metalloproteinase isolated from Bothrops jararaca venom, is a potent hemorrhagic toxin. A cDNA encoding HF3 indicated that it is a multidomain molecule composed of a pro domain, a catalytic domain with a zinc binding sequence, followed by disintegrin like and cysteine-rich domains. It is known that metalloproteinases play a relevant role in the pathogenesis of venom-induced local tissue damage including inflammation. In this study we evaluated the effects of native HF3 and its recombinant disintegrin-like/cysteine-rich domains (DC-HF3) on alpha(M)beta(2) mediated phagocytosis of opsonized-zymosan particles by macrophages. HF3 and DC HF3 significantly increased phagocytosis and this activity was inhibited by anti alpha(M) and anti-beta(2) antibodies. The data show the ability of P-III metalloproteinases to activate macrophages for phagocytosis through integrin alpha(M)beta(2) and suggest that the disintegrin-like/cysteine-rich domains are important for this effect. This is the first report on the activation of phagocytosis via alpha(M)beta(2) integrin by a metalloproteinase containing disintegrin-like/cysteine-rich domains. PMID- 15336557 TI - ADRP is dissociated from lipid droplets by ARF1-dependent mechanism. AB - Adipocyte differentiation-related protein (ADRP) is a member of PAT proteins existing in lipid droplets (LDs). By yeast two-hybrid screening, we identified ADP-ribosylation factor 1 (ARF1) as a binding partner of ADRP. The interaction of ADRP and ARF1 was verified by GST pull-down and co-immunoprecipitation experiments. Interestingly, ADRP precipitated the GDP-bound ARF1 preferentially to the GTP-bound ARF1. Consistent with this, either brefeldin A (BFA), a fungal metabolite to inhibit ARF-GEF, or a dominant-negative mutant of ARF1 caused dissociation of ADRP from LD. On the other hand, overexpression of wild-type ARF1 did not promote the ADRP dissociation or new LD formation. By using deletion mutants, a central domain of ADRP, which is dispensable for LD binding, was shown to bind to ARF1. The present study showed that the GDP-bound ARF1 induces dissociation of ADRP from the LD surface, and that LD is a target of BFA action. PMID- 15336558 TI - Relation between tRNase activity and the structure of colicin D according to X ray crystallography. AB - Colicin D is a plasmid-encoded proteinaceous toxin which kills sensitive Escherichia coli. Toxicity stems from ribonuclease activity that targets exclusively four isoacceptors of tRNA(Arg) with a cleavage position between 38 and 39 of the corresponding anticodons. Since no other tRNAs with the same sequences at 38 and 39 as tRNA(Arg)s are cleaved, colicin D should be capable of recognizing some higher order structure of tRNAs. We report here two crystal structures of catalytic domains of colicin D which have different N-terminal lengths, both complexed with its cognate inhibitor protein, ImmD. A row of positive charge patches is found on the surface of the catalytic domain, suggestive of the binding site of the tRNAs. This finding, together with our refined tRNase activity experiments, indicates that the catalytic domain starting at position 595 has activity almost equivalent to that of colicin D. PMID- 15336559 TI - Acetylcholine esterase protects LDL against oxidation. AB - Acetylcholine esterase (AChE) and paraoxonase 1 (PON1) are both serum ester hydrolases, which are associated with the prevalence of myocardial infarction. Both genes are located in close proximity on chromosome 7q21-22. As PON1 was suggested to protect against cardiovascular diseases secondary to its ability to break down oxidized lipids and to inhibit LDL oxidation, we examined AChE capacity to protect LDL against oxidation. Preincubation of LDL with AChE retarded the onset of copper ion-induced LDL oxidation in a concentration dependent manner. AChE significantly reduced the formation of lipid peroxides and TBARS during the course of LDL oxidation, by up to 45%. This effect was associated with AChE-mediated hydrolysis of lipid peroxides, which accounts for the inhibition in the onset of LDL oxidation, the oxidative propagation phase, and aldehyde formation. We conclude that AChE, similar to PON1, can hydrolyze lipid peroxides and thus may prevent the accumulation of oxidized LDL and attenuate atherosclerosis development. PMID- 15336560 TI - Down-regulation of beta2-adrenergic receptor expression by exercise training increases IL-12 production by macrophages following LPS stimulation. AB - Three-week exercise training decreased the steady state level of beta(2) adrenergic receptor (beta(2)AR) mRNA in peritoneal macrophages from BALB/c mice. When peritoneal macrophages from both exercise-trained and sedentary control mice were stimulated with lipopolysaccharide (LPS), interleukin (IL)-12 mRNA and protein expression was markedly higher in trained mice than in control mice. To determine whether enhanced production of IL-12 was associated with decreased expression of beta(2)AR, we transfected the macrophage cell line, RAW264, with a eukaryotic expression vector containing beta(2)ar cDNA, establishing a cell line overexpressing beta(2)AR (RAWar). Following LPS stimulation, IL-12 mRNA and protein expression was significantly lower in RAWar cells than in RAW264 cells transfected with vector alone (RAWvec). Furthermore, when the expression of transfected beta(2)AR in RAWar cells was down-regulated by a tetracycline repressor-regulated mammalian expression system, expression of IL-12 mRNA and protein following LPS stimulation tended to return to the levels in RAWvec cells. These findings indicate that macrophage production of IL-12 following LPS stimulation is regulated by the expression level of beta(2)AR, suggesting that the down-regulation of beta(2)AR expression associated with exercise training improves IL-12-induced type 1 helper T cell-mediated immune responses. PMID- 15336561 TI - TcRRMs and Tcp28 genes are intercalated and differentially expressed in Trypanosoma cruzi life cycle. AB - The identification and characterization of RNA binding proteins in Trypanosoma cruzi are particularly relevant as they play key roles in the regulatory mechanisms of gene expression. In this work, we have identified coding sequences for the proteins, named TcRRM1 and TcRRM2, in the EST database generated by the T. cruzi genomic initiative. TcRRM1 and TcRRM2 contain two RNA binding domains (RRM) and are very similar to two Trypanosoma brucei RNA binding proteins previously reported, Tbp34 and Tbp37, and to a not yet annotated ORF in Leishmania major genome project. The T. cruzi RRM genes are organized in tandem, alternating with copies of Tcp28, a gene of unknown function. However, TcRRM transcript accumulation is higher in the spheromastigote stage, while Tcp28 transcripts accumulate more in the trypomastigote stage suggesting developmental regulation. PMID- 15336562 TI - Measurement of DNA helical change for the binding of cyclic AMP receptor protein to lac DNA. AB - Cyclic AMP receptor protein (CRP) binds to DNA and induces DNA bending. DNA bending plays an important role in transcription activation of many genes in Escherichia coli. To know DNA helical change by CRP, we used molar cyclization factor (j(M)), which is sensitive to helical twist or torsional alignment. Six consecutive DNA fragments were constructed that contained phasing sequence at left arm and CRP binding site in center. j(M) values measured in this study were approximately 10 times increased by CRP relative to that measured for DNA only. The analysis of set of j(M) values indicated that CRP induced DNA bending without the helical change like helical twisting or torsional alignment change. PMID- 15336563 TI - Proteins are polyisoprenylated in Arabidopsis thaliana. AB - Isoprenoid lipids were found to be covalently linked to proteins of Arabidopsis thaliana. Their identity (polyprenols: Prenol-9-11 with Pren-10 dominating and dolichols: Dol-15-17 with Dol-16 dominating) was confirmed by means of HPLC/ESI MS with application of the multiple reaction monitoring technique as well as metabolic labeling of Arabidopsis plants with [(3)H]mevalonate and other precursors. The occurrence of typical farnesol-, geranylgeraniol-, and phytol modified proteins was also noted. Radioisotopic labeling allowed detection of several proteins that were covalently bound to mevalonate-derived isoprenoid alcohols. A significant portion of polyisoprenylated proteins was recovered in the cytosolic/light vesicular fraction of Arabidopsis cells upon subfractionation. Taken together our data prove that a subset of plant proteins is polyisoprenylated. PMID- 15336564 TI - A STAT3 dimer formed by inter-chain disulphide bridging during oxidative stress. AB - Signal transducer and activator of transcription (STAT) proteins are activated by cytokines and growth factors to play distinct roles in immune responses and developmental processes. STATs were thought to exist as latent, cytoplasmic monomers and activation to require dimer formation was mediated exclusively by reciprocal phospho-tyrosine/SH2-domain interactions, but recent evidence of cytoplasmic STAT complexes, including dimers, and unphosphorylated STATs in the nucleus has challenged these notions. STAT complexes detected by conventional SDS PAGE, including a STAT3 dimer, have been reported. We show that such complexes can form during cell lysis and be disrupted with DTT, suggesting inter-chain disulphide bridging. STAT3 also forms a related complex in cells upon oxidative stress. We map the interaction to the amino-terminal domain of STAT3 and use mass spectrometry to implicate cysteine 259 as the reactive residue. The redox sensitivity of STAT3 may be significant, given its activation in cells in response to reactive oxygen species. PMID- 15336565 TI - Expression of methionine aminopeptidase 2, N-myristoyltransferase, and N myristoyltransferase inhibitor protein 71 in HT29. AB - Protein myristoylation is a co-translational process, catalyzed by N myristoyltransferase (NMT) that occurs after the initiating methionine is removed by methionine aminopeptidase (MetAP). The enzymes NMT and MetAP play a major role in the process of myristoylation of oncoproteins including the c-src family. In this study, we examined the levels of expression of MetAP2, NMT, and NMT inhibitor protein 71 (NIP71) in human colon cancer cell lines (HCCLs). We examined the influence of cell density on the expression of the above proteins in HT29 cells. Western blot analysis of MetAP2 and NMT demonstrated higher levels of protein expression in low density of HT29 while low expression in high density was observed. In addition, we observed that NIP71 and pp60(c-src) expressions were dependent on the cell density of HT29. This is the first study demonstrating the expression of MetAP2, NMT, pp60(c-src), and NIP71 in HCCLs. PMID- 15336566 TI - Formation of radical cations in a model for the metabolism of aromatic hydrocarbons. AB - To test the hypothesis that electrophilic radical cations are the major ultimate electrophilic and carcinogenic forms of benz[a]anthracene (BA), dibenz[a,h]anthracene (DBA), and benzo[a]pyrene (BP), we have focused on a chemical model of metabolism which parallels and duplicates known or potential metabolites of some polycyclic hydrocarbons formed in cells. Studies of this model system show that radical cations are hardly formed, if at all, in the case of BA or DBA but are definitely formed in the cases of the carcinogen BP as well as the non-carcinogenic hydrocarbons, pyrene and perylene. We conclude that the carcinogenicities of BA, DBA, BP, pyrene, and perylene are independent of one electron oxidation to radical cation intermediates. PMID- 15336567 TI - A novel TNFalpha antagonizing peptide-Fc fusion protein designed based on CDRs of TNFalpha neutralizing monoclonal antibody. AB - The variable regions of antibody molecules bind antigens with high affinity and specificity. The binding sites are imparted largely to the hypervariable portions (i.e., CDRs) of the variable region. Peptides derived from CDRs can bind antigen with similar specificity acting as mimic of antibody and become drug-designing core, although with markedly lower affinity. In order to increase the affinity and bioactivity, in this study, a novel peptide (PT) designed on CDRs of a TNFalpha neutralizing monoclonal antibody Z12 was linked with Fc fragment of human IgG1. The interaction mode of PT-linker-Fc (PLF) with TNFalpha was analyzed with computer-guided molecular modeling method. After expression in Escherichia coli and purification, recombinant PT-linker-Fc could bind directly with the TNFalpha coated on the ELISA plates. Furthermore, PLF could competitively inhibit the binding of Z12 to TNFalpha and also inhibit the TNFalpha-induced cytotoxicity on L929 cells. The TNFalpha antagonizing activity of PLF was significantly higher than that of the free peptide. This study highlights the potential of human Fc to enhance the potency of peptides designed on the CDRs of antibodies and could be useful in developing new TNFalpha antagonists. PMID- 15336568 TI - Thermal inactivation of protective antigen of Bacillus anthracis and its prevention by polyol osmolytes. AB - Protective antigen (PA) of Bacillus anthracis is the main immunogen of all anthrax vaccines. It is a highly thermolabile molecule and loses its activity rapidly when exposed to higher temperatures. Earlier some cosolvents had been used to stabilize PA with variable success but no study has been done to find out the primary cause of PA thermal inactivation. This study aims at elucidating the predominant cause of thermal inactivation of PA in order to develop more effective strategies for its thermostabilization. The prime cause for the loss of biological activity of PA at high temperature was its aggregation and an inverse correlation between PA activity and its aggregation on heating was observed. Inactivation of the protein by autolysis did not occur. This paper reports the use of a series of polyol osmolytes to stabilize PA. Different polyols stabilized PA to a different extent against thermal inactivation in a concentration dependent manner, with glycerol stabilizing to the maximum extent. Addition of NaCl to glycerol solution further enhanced the thermal stability of PA. An increase in the T(1/2) value, the temperature at which 50% of the activity is retained during short-term incubation, of more than 20 degrees C was observed. The half-life (t(1/2)) of PA thermal inactivation at 40 degrees C increased by more than 6 times in the presence of the mixture of glycerol and NaCl as compared to control. This study demonstrates for the first time that aggregation of the PA molecule is the predominant cause of its thermal inactivation, and can be very effectively prevented by the use of glycerol and other polyols to increase the shelf life of the recombinant vaccine against anthrax. PMID- 15336569 TI - Whole genome analysis reveals a high incidence of non-optimal codons in secretory signal sequences of Escherichia coli. AB - Translational pausing may occur due to a number of mechanisms, including the presence of non-optimal codons, and it is thought to play a role in the folding of specific polypeptide domains during translation and in the facilitation of signal peptide recognition during sec-dependent protein targeting. In this whole genome analysis of Escherichia coli we have found that non-optimal codons in the signal peptide-encoding sequences of secretory genes are overrepresented relative to the "mature" portions of these genes; this is in addition to their overrepresentation in the 5'-regions of genes encoding non-secretory proteins. We also find increased non-optimal codon usage at the 3' ends of most E. coli genes, in both non-secretory and secretory sequences. Whereas presumptive translational pausing at the 5' and 3' ends of E. coli messenger RNAs may clearly have a general role in translation, we suggest that it also has a specific role in sec dependent protein export, possibly in facilitating signal peptide recognition. This finding may have important implications for our understanding of how the majority of non-cytoplasmic proteins are targeted, a process that is essential to all biological cells. PMID- 15336570 TI - OGT functions as a catalytic chaperone under heat stress response: a unique defense role of OGT in hyperthermia. AB - Protein O-GlcNAcylation is proceeded by O-linked GlcNAc transferase (OGT) in nucleocytoplasm and is involved in many biological processes although its physiological role is not clearly defined. To identify the functional significance of O-GlcNAcylation, we investigated heat stress effects on protein O GlcNAcylation. Here, we found that protein O-GlcNAcylation was significantly increased in vivo during acute heat stress in mammalian cells and simultaneously, the enhanced protein O-GlcNAcylation was closely associated with cell survival in hyperthermia. Our results demonstrate that hyperthermal cytotoxicity may considerably be facilitated under the condition of insufficient level of protein O-GlcNAcylation inside cells. Furthermore, OGT reaction might be crucial for triggering thermotolerance to recover hyperthermal sensitivity without particular induction of heat shock proteins (hsps). Thus, we propose that OGT can respond rapidly to heat stress through the enhancement of nucleocytoplasmic protein O GlcNAcylation for a rescue from the early phase of hyperthermal cytotoxicity. PMID- 15336571 TI - Chemokine CCL20 enhances the growth of HuH7 cells via phosphorylation of p44/42 MAPK in vitro. AB - CCR6 is the receptor of chemokine CCL20. In the present study, we demonstrated that the surface expression of CCR6 was enhanced on the human HCC cell lines (HuH7, PLC/PRF/5, and HepG2) especially on HuH7 cells, but not on HLE or HLF cells. These HCC cell lines (HuH7, PLC/PRF/5, and HepG2) especially the HuH7 cells secreted a significant amount of CCL20 spontaneously, whereas HLE or HLF did not. Stimulation by CCL20 up-regulated the mRNA expression of CCR6 in HuH7 cells and significantly enhanced the growth of HuH7 cells. CCL20-stimulated growth of HuH7 cells was abrogated by the inhibition of downstream signal transduction pathway mediated by p44/42 MAPK, but not by p38 MAPK or SAPK/JNK. CCR6 expression in human HCC tissues was confirmed by RT-PCR. These results indicate that the growth of a proportion of human HCC cells may be mediated by CCL20-CCR6 axis, like HuH7 cells, in an autocrine or paracrine manner. PMID- 15336572 TI - Functional evidence for a supramolecular structure for the Streptomyces lividans potassium channel KcsA. AB - Here we present functional evidence for involvement of poly-(R)-3-hydroxybutyrate (PHB) and inorganic polyphosphate (polyP) in ion conduction and selection at the intracellular side of the Streptomyces lividans potassium channel, KcsA. At < or = 25 degrees C, KcsA forms channels in planar bilayers that display signal characteristics of PHB/polyP channels at the intracellular side; i.e., a preference for divalent Mg(2+) cations at pH 7.2, and a preference for monovalent K+ cations at pH 6.8. Between 25 and 26 degrees C, KcsA undergoes a transition to a new conformation in which the channel exhibits high selectivity for K+, regardless of solution pH. This suggests that basic residues of the C-terminal polypeptides have moved closer to the polyP end unit, reducing its negative charge. The data support a supramolecular structure for KcsA in which influx of ions is prevented by the selectivity pore, whereas efflux of K+ is governed by a conductive core of PHB/polyP in partnership with the C-terminal polypeptide strands. PMID- 15336573 TI - Prostanoid EP4 receptor is involved in suppression of 3T3-L1 adipocyte differentiation. AB - Prostaglandins (PGs) have been shown to play various roles in adipogenesis. In this study, we investigated on which PGE receptor subtypes are involved in the inhibition of 3T3-L1 preadipocyte differentiation. The triglyceride content of cells, used as an index of differentiation, was decreased when PGE(2), the FP agonist fluprostenol or dibutyryl cAMP, was exogenously added to differentiation cocktails. 3T3-L1 preadipocyte cells express mRNAs for the prostanoid EP4, FP, and IP receptors. PGE(2) and the EP4 agonist AE1-329 increased cAMP levels in preadipocytes in a dose-dependent manner. AE1-329 suppressed the expression induction of differentiation marker genes such as resistin and peroxisome proliferator-activated receptor-gamma. The inhibitory effect of PGE(2) but not that of fluprostenol was reversed by the addition of the EP4 antagonist AE3-208. AE3-208 mimicked the differentiation-promoting effects of indomethacin. These results suggest that the EP4 receptor mediates the suppressive action of PGE(2) in 3T3-L1 adipocyte differentiation. PMID- 15336574 TI - MGluRs regulate the expression of neuronal calcium sensor proteins NCS-1 and VILIP-1 and the immediate early gene arg3.1/arc in the hippocampus in vivo. AB - The metabotropic glutamate receptor (mGluR) agonist (R,S)-3,5 dihydroxyphenylglycine (DHPG) is involved in several forms of hippocampal synaptic plasticity. DHPG application can induce slow-onset potentiation, a form of long-term potentiation (LTP), in the dentate gyrus and in the CA1 region in vivo. The induction of LTP correlates with increased expression levels of neuronal calcium sensor (NCS), considered as key elements for plasticity. In this study we investigated mGluR- and time-dependent changes in the expression of two different NCS proteins. Following DHPG application in vivo NCS-1 and VILIP-1 expression increased, with significant levels reached after 8 and 24h. The effect was attenuated by treatment with the group I mGluR specific antagonist S-4 carboxyphenylglycine. The immediate early gene (IEG) arg3.1/arc showed highest expression levels 2h after DHPG-treatment. Therefore, mGluRs at concentrations which induce synaptic plasticity regulate the expression of IEGs and NCS proteins in different time frames and thus contribute to late phases of synaptic plasticity. PMID- 15336575 TI - Leptin and insulin down-regulate angiopoietin-like protein 3, a plasma triglyceride-increasing factor. AB - We reported previously that angiopoietin-like protein3 (ANGPTL3), a liver specific secretory factor, increased plasma triglyceride (TG) via inhibition of lipoprotein lipase and free fatty acid (FFA) by activating adipose-lipolysis. The current study examined the regulation of Angptl3 by leptin and insulin, both of which are key players in the metabolic syndrome. Angptl3 expression and plasma ANGPTL3 levels were increased in leptin-resistant C57BL/6J(db/db) and -deficient C57BL/6J(ob/ob) mice, relative to the control. Leptin supplements decreased Angptl3 gene expression and plasma ANGPTL3 in C57BL/6J(ob/ob) mice. The changes of Angptl3 were associated with alterations of plasma TG and FFA levels. Leptin treatment directly suppressed Angptl3 gene expression in hepatocytes. Angptl3 gene expression and plasma protein levels were also increased in insulin deficient streptozotocin-treated mice. Insulin treatment of hepatocytes decreased Angptl3 gene expression and protein secretion. Our results suggest that elevated ANGPTL3 by leptin- or insulin-resistance is attributed to increased plasma TG and FFA concentrations in obesity. PMID- 15336576 TI - Leishmania donovani activates nuclear transcription factor-kappaB in macrophages through reactive oxygen intermediates. AB - Interaction of Leishmania donovani with macrophages antagonizes host defense mechanisms by interfering with a cascade of cell signaling processes in the macrophages. An early intracellular signaling event that follows receptor engagement is the activation of transcription factor NF-kappaB. It has been reported earlier that NF-kappaB-dependent signaling pathway regulates proinflammatory cytokine release. We therefore investigated the effect of L. donovani infectivity on this nuclear transcription factor in macrophage cell line J774A.1. Both L. donovani and its surface molecule lipophosphoglycan (LPG) resulted in a dose- and time-dependent activation of NF-kappaB-DNA binding activity in an electrophoretic mobility shift assay. We also report the involvement of IkappaB-alpha and IkappaB-beta in the persistent activation of NF kappaB by L. donovani. We demonstrate that the NF-kappaB activation was independent of viability of the parasite. Electrophoretic mobility supershift assay indicated that the NF-kappaB complex consists of p65 and c-rel subunits. The interaction of parasite with the macrophages and not the cellular uptake was important for NF-kappaB activation. Both p38 and ERK mitogen activated protein kinase (MAP) activation appears to be necessary for NF-kappaB activation by LPG. Preincubation of cells with antioxidants resulted in inhibition of L. donovani induced NF-kappaB activation, thereby suggesting a potential role of reactive oxygen species in L. donovani induced intracellular signaling. The present data indicate that antioxidants could play an important role in working out various therapeutic modalities to control leishmaniasis. PMID- 15336577 TI - Efficacy of subcutaneous insulin lispro versus continuous intravenous regular insulin for the treatment of patients with diabetic ketoacidosis. AB - PURPOSE: To compare the efficacy and safety of subcutaneous insulin lispro with that of a standard low-dose intravenous infusion protocol of regular insulin in patients with uncomplicated diabetic ketoacidosis. METHODS: In this prospective, randomized open trial, 20 patients treated with subcutaneous insulin lispro were managed in regular medicine wards (n=10) or an intermediate care unit (n=10), while 20 patients treated with the intravenous protocol were managed in the intensive care unit. Patients treated with subcutaneous lispro received an initial injection of 0.3 unit/kg followed by 0.1 unit/kg/h until correction of hyperglycemia (blood glucose levels <250 mg/dL), followed by 0.05 to 0.1 unit/kg/h until resolution of diabetic ketoacidosis (pH > or =7.3, bicarbonate > or =18 mEq/L). Patients treated with intravenous regular insulin received an initial bolus of 0.1 unit/kg, followed by an infusion of 0.1 unit/kg/h until correction of hyperglycemia, then 0.05 to 0.1 unit/kg/h until resolution of diabetic ketoacidosis. RESULTS: Mean (+/- SD) admission biochemical parameters in patients treated with subcutaneous lispro (glucose: 674 +/- 154 mg/dL; bicarbonate: 9.2 +/- 4 mEq/L; pH: 7.17 +/- 0.10) were similar to values in patients treated with intravenous insulin (glucose: 611 +/- 264 mg/dL; bicarbonate: 10.6 +/- 4 mEq/L; pH: 7.19 +/- 0.08). The duration of treatment until correction of hyperglycemia (7 +/- 3 hours vs. 7 +/- 2 hours) and resolution of ketoacidosis (10 +/- 3 hours vs. 11 +/- 4 hours) in patients treated with subcutaneous lispro was not different than in patients treated with intravenous regular insulin. There were no deaths in either group, and there were no differences in the length of hospital stay, amount of insulin until resolution of diabetic ketoacidosis, or in the rate of hypoglycemia between treatment groups. Treatment of diabetic ketoacidosis in the intensive care unit was associated with 39% higher hospitalization charges than was treatment with subcutaneous lispro in a non-intensive care setting ($14,429 +/- $5243 vs. $8801 +/- $5549, P <0.01). CONCLUSION: Treatment of adult patients who have uncomplicated diabetic ketoacidosis with subcutaneous lispro every hour in a non intensive care setting may be safe and more cost-effective than treatment with intravenous regular insulin in the intensive care unit. PMID- 15336578 TI - The association between quality improvement activities performed by managed care organizations and quality of care. AB - PURPOSE: Little data are available to assess the efforts of managed care organizations to improve quality of care. This analysis assessed differences in performance rates between organizations with and without quality improvement activities. METHODS: We reviewed 399 self-reported quality improvement activities submitted by organizations seeking accreditation by the National Committee for Quality Assurance. Processes or outcomes assessed in quality improvement activities were linked to corresponding measures in the effectiveness-of-care database of the Health Plan Employer Data and Information Set (HEDIS). Performance rates for managed care organizations with and without quality improvement activities were then compared. RESULTS: The cross-sectional analysis included 79 quality improvement activities from 50 organizations, covering 12 effectiveness-of-care categories. Each activity had a matching performance score in the database. Financial incentives for providers were associated with substantially higher performance rates in organizations employing this type of intervention. Eight effectiveness-of-care categories had at least four organizations reporting specific quality improvement activities for the care category of interest; statistically significant improvements were observed for follow-up visits for patients after hospitalization for mental illness, checkups after delivery, and screening for cervical cancer. CONCLUSION: Based on objective and audited information, the estimated effects of self-reported quality improvement activities were often small and inconsistent. In some instances, the observed effect was contrary to the expected direction. Limitations of the available dataset and the caveats of a cross-sectional study design precluded a number of analytical options. Longer-term, prospective studies are needed to explore further the relation between quality improvement activities and objective measures of clinical performance. PMID- 15336579 TI - Patients admitted to hospital with suspected pneumonia and normal chest radiographs: epidemiology, microbiology, and outcomes. AB - PURPOSE: To describe the prevalence of patients admitted to hospital with a diagnosis of community-acquired pneumonia who have normal chest radiographs; the extent to which patients actually had pneumonia on radiographs; and to compare presentation and outcomes in patients with a lower respiratory tract infection and those whose clinical diagnosis of pneumonia was confirmed by radiography. METHODS: We studied a population-based cohort of 2706 adults who were admitted with suspected pneumonia and managed using a clinical pathway. We stratified patients by presence or absence of radiograph-confirmed pneumonia, and compared their characteristics and in-hospital mortality. We also performed an independent review of a 10% sample of "normal" chest radiographs and classified them according to the presence or absence of pneumonia. RESULTS: One third (n=911) of patients admitted with pneumonia had their initial radiograph reported as "no pneumonia." Independent review found that only 7% (6/92) of radiographs developed an opacity that confirmed pneumonia. Characteristics were similar among admitted patients irrespective of radiographic findings, although patients without pneumonia on radiograph were older (mean [+/- SD] age, 73 +/- 15 years vs. 68 +/- 19 years, P <0.001) and had greater pneumonia-specific severity-of-illness scores (104 +/- 32 vs. 99 +/- 37, P=0.004). Patients without radiographic confirmation of pneumonia had similar rates of positive sputum cultures (32% [87/271] vs. 30% [208/706], P=0.42) and blood cultures (6% [35/576] vs. 8% [100/1241], P=0.13), but microbiology results differed, with a shift away from Streptococcus pneumoniae towards other streptococci species and gram-negative aerobic bacilli. In-hospital mortality was similar for both groups of patients (8% [64/911] in the unconfirmed pneumonia group vs. 10% [165/1795] in the confirmed group, adjusted P=0.09). CONCLUSION: One third of patients suspected of having pneumonia and admitted to hospital did not have pneumonia, but had serious lower respiratory tract infections with substantial rates of bacteremia and mortality. The absence of radiographic findings should not supercede clinical judgment and empiric treatments in these patients. PMID- 15336580 TI - Insulin-like growth factor 1 as a predictor of ischemic stroke outcome in the elderly. AB - PURPOSE: To examine whether serum insulin-like growth factor 1 (IGF-1) and IGF binding protein 3 (IGFBP-3) concentrations, determined early after the onset of stroke, are predictive of clinical outcome in elderly patients. METHODS: The sample comprised 85 patients (mean [+/- SD] age, 83 +/- 7.4 years; range, 67 to 99 years; 34% male) who were admitted with acute stroke to a geriatric ward between January 1998 and June 2000, and 88 control patients who were similar in age and sex. Clinical and laboratory assessments, computed tomographic scan of the head, carotid ultrasonography, and electrocardiography were employed to define the clinical and etiologic stroke subtype. Fasting blood samples were collected within 24 hours of admission for IGF-I and IGFBP-3 measurement. Univariate and multiple logistic regression analyses, with adjustment for other related clinical covariates, were used to assess the relation of IGF-I and IGFBP 3 to poor outcome, defined as severe disability (Barthel index <60/100) or death, at 1 month (or at discharge), 3 months, and 6 months. RESULTS: Mean (+/- SD) IGF 1 levels were lower in patients with stroke than in controls (69 +/- 45 ng/mL vs. 102 +/- 67 ng/mL, P adjusted for age = 0.001). The mean IGF-1/IGFBP-3 molar ratio was also lower in stroke patients (0.12 +/- 0.07 vs. 0.19 +/- 0.09, P adjusted for age <0.0001). However, there was no relation of hormone levels to either the clinical subtype of stroke or the extent of neurologic impairment. IGF-1 levels were inversely related to poor outcome (mainly death) at 3 and 6 months, independent of other clinical covariates that were highly predictive of outcome, such as age and stroke scale score on admission (hazard ratio for death at 6 months for each 20-ng/mL increase = 0.7; 95% confidence interval: 0.5 to 0.9). An independent association of the molar ratio with death at 3 and 6 months was also found. CONCLUSION: Low levels of circulating IGF-1 may predict the clinical outcome of stroke in elderly patients. PMID- 15336581 TI - Improving completion of advance directives in the primary care setting: a randomized controlled trial. AB - BACKGROUND: Since 1991, hospitals have asked patients whether they have advance directives, but few patients complete these documents. We assessed two simple interventions to improve completion of advance directives among elderly or chronically ill outpatients. METHODS: We conducted a cluster randomized controlled trial involving 1079 patients from five general medicine clinics that were affiliated with an academic medical center. Patients were either > or =70 years of age or > or =50 years old with a chronic illness. The study comprised three arms: physician reminders recommending documentation of advance directives, physician reminders plus mailing advance directives to patients together with educational literature, or neither intervention (control). The main outcome measure was completion of an advance directive. RESULTS: After 28 weeks, 1.5% (5/332) of patients in the physician reminder group, 14% (38/277) in the physician reminder plus patient mailing group, and 1.8% (5/286) in the control group had completed advance directives. In multivariate analyses, patients in the physician reminder plus patient mailing group were much more likely than controls to have completed advance directives (odds ratio [OR] = 5.9; 95% confidence interval [CI]: 1.5 to 22), whereas patients in the physician reminder-only group were no more likely than controls to have completed advance directives (OR = 0.88; 95% CI: 0.21 to 3.7). CONCLUSION: Mailing health care proxy and living will forms and literature to patients before an appointment at which their physicians received a reminder about advance directives yielded a small but significant improvement in completion of these documents. A physician reminder alone did not have an effect. PMID- 15336582 TI - Magnesium prophylaxis for arrhythmias after cardiac surgery: a meta-analysis of randomized controlled trials. AB - BACKGROUND: Magnesium supplementation may reduce the incidence of arrhythmias, which often occur after cardiac surgery; however, recent findings of the effectiveness of magnesium prophylaxis have yielded discrepant results. METHODS: We searched electronic databases for randomized controlled trials of magnesium for the prevention of arrhythmias after cardiac surgery. The primary outcomes comprised the incidence of supraventricular and ventricular arrhythmias, and the secondary outcomes comprised serum magnesium concentration, length of hospital stay, myocardial infarction, and mortality. Effect sizes were estimated using a random-effects model. RESULTS: Seventeen trials (n=2069 patients) met the inclusion criteria. Pooled serum magnesium concentration at 24 hours after surgery in the treatment group was significantly higher than that in the control group (weighted mean difference=0.45 mmol/L [1.1 mg/dL]; 95% confidence interval [CI]: 0.30 to 0.59 mmol/L [0.7 to 1.4 mg/dL]; P <0.001). Magnesium supplementation reduced the risk of supraventricular arrhythmias (relative risk [RR]=0.77; 95% CI: 0.63 to 0.93; P=0.002) and ventricular arrhythmias (RR = 0.52; 95% CI: 0.31 to 0.87; P <0.0001), but had no effect on the length of hospital stay (weighted mean difference=-0.28 days; 95% CI: -0.70 to 1.27 days; P=0.48), the incidence of perioperative myocardial infarction (RR=1.03; 95% CI: 0.52 to 2.05; P = 0.99), or mortality (RR=0.97; 95% CI: 0.43 to 2.20; P=0.94). CONCLUSION: Administration of prophylactic magnesium reduced the risk of supraventricular arrhythmias after cardiac surgery by 23% (atrial fibrillation by 29%) and of ventricular arrhythmias by 48%. Supplementation had no notable benefit with respect to length of hospitalization, incidence of myocardial infarction, or mortality. PMID- 15336583 TI - Bedside diagnosis of coronary artery disease: a systematic review. AB - PURPOSE: To assess the accuracy of bedside findings for diagnosing coronary artery disease and acute myocardial infarction. METHODS: A MEDLINE search was performed to retrieve articles published from January 1966 to January 2003 that were relevant to the bedside diagnosis of coronary disease in adults. RESULTS: In patients with stable, intermittent chest pain, the most useful bedside predictors for a diagnosis of coronary disease were found to be the presence of typical angina (likelihood ratio [LR]=5.8; 95% confidence interval [CI]: 4.2 to 7.8), serum cholesterol level >300 mg/dL (LR=4.0; 95% CI: 2.5 to 6.3), history of prior myocardial infarction (LR=3.8; 95% CI: 2.1 to 6.8), and age >70 years (LR=2.6; 95% CI: 1.8 to 4.0). Nonanginal chest pain (LR=0.1; 95% CI: 0.1 to 0.2), pain duration >30 minutes (LR=0.1; 95% CI: 0.0 to 0.9), and intermittent dysphagia (LR=0.2; 95% CI: 0.1 to 0.8) argued against a diagnosis of coronary disease. In patients with acute chest pain, the most important bedside predictors for a diagnosis of myocardial infarction were new ST elevation (LR=22; 95% CI: 16 to 30), new Q waves (LR=22; 95% CI: 7.6 to 62), and new ST depression (LR=4.5; 95% CI: 3.6 to 5.6). A normal electrocardiogram (LR=0.2; 95% CI: 0.1 to 0.3), chest wall tenderness (LR=0.3; 95% CI: 0.2 to 0.4), and pain that was pleuritic (LR=0.2; 95% CI: 0.2 to 0.3), sharp (LR=0.3; 95% CI: 0.2 to 0.5), or positional (LR=0.3; 95% CI: 0.2 to 0.5) argued against the diagnosis of myocardial infarction. CONCLUSION: The accuracy of bedside predictors depends on the clinical setting. In the evaluation of stable, intermittent chest pain, a patient's description of pain was found to be the most important predictor of underlying coronary disease. In the evaluation of acute chest pain, the electrocardiogram was the most useful bedside predictor for a diagnosis of myocardial infarction. Aside from the extremes in cholesterol values, the analysis of traditional risk factors changed the probability of coronary disease or myocardial infarction very little or not at all. PMID- 15336584 TI - Hepatitis C treatment update. AB - Hepatitis C is a leading cause of chronic liver disease in the United States, and the prevalence of hepatitis C-associated complications is increasing. Therapy with pegylated interferon and ribavirin has become the standard of care for chronic hepatitis C; the sustained response rate for treatment-naive patients is about 55%. If certain patients fail to achieve a 12-week treatment milestone, an early virologic response, they may be taken off treatment early, potentially sparing them from unnecessary medication. Adherence is critical for treatment success. Although side effects continue to be a hindrance to the success of therapy, agents such as growth factors and antidepressants may help patients to maintain medication dosing and complete treatment. Therapy is generally recommended for those in whom the infection is most likely to progress to cirrhosis; however, there is continued debate about the suitability of certain patients for treatment, including those with persistently normal aminotransferase levels or acute hepatitis C and nonresponders to conventional treatment. Four broad groups of investigational therapeutic agents appear promising for future therapy: modified interferons and ribavirins, immunomodulators, viral life-cycle targets, and antifibrotic agents. PMID- 15336585 TI - Cases from the Osler Medical Service at Johns Hopkins University. Diagnosis: P. carinii pneumonia and primary pulmonary sporotrichosis. AB - PRESENTING FEATURES: A 53-year-old man who had human immunodeficiency virus (HIV) presented to the Johns Hopkins Hospital with a 3-month history of increasing dysphagia, cough, dyspnea, chest pain, and an episode of syncope. His past medical history was notable for oral and presumptive esophageal candidiasis that was treated with fluconazole 6 months prior to presentation. Three months prior to presentation, he discontinued his medications, and his symptoms of dysphagia recurred. During that time he developed intermittent fevers and chills, progressively worsening dyspnea on exertion, and a cough productive of white sputum. He also reported a 40-lb weight loss over the past 3 months. On the day prior to presentation, he had chest pain and shortness of breath followed by weakness, dizziness, and a brief syncopal episode. He denied orthopnea, paroxysmal nocturnal dyspnea, lower extremity edema, jaundice, hemoptysis, hematemesis, melena, hematochezia, or diarrhea. There was no history of alcohol use, and he stopped smoking tobacco approximately 1 month previously. He smoked cocaine but denied injection drug use. The patient had never been on antiretroviral therapy and had never had his CD4 count or viral load measured. On physical examination, the patient was a thin, cachectic man who appeared older than his stated age. His vital signs were notable for blood pressure of 102/69 mm Hg, resting tachycardia of 102 beats per minute, resting oxygen saturation of 92% on room air, normal resting respiratory rate, and a temperature of 38.1 degrees C. His oropharynx was clear, with no signs of thrush or mucosal ulcers. His pulmonary examination was notable for diminished breath sounds in the lower lung fields bilaterally. Cardiac, abdominal, and neurologic examinations were normal. His skin was intact, with no visible petechiae, rashes, nodules, or ulcers. Laboratory studies showed a total white blood cell count of 3.2 x 10(3)/microL, with a total lymphocyte count of 330/microL, hematocrit of 30.2%, a serum sodium level of 129 mEq/L, and a serum lactate dehydrogenase level of 219 IU/L. The patient had an absolute CD4 count of 8 cells/mm3 and a HIV viral load of 86,457 copies/mL. His arterial blood gas on room air had a pH of 7.51, a PCO2 of 33 mm Hg, and a PO2 of 55 mm Hg. Electrocardiogram and serial serum cardiac enzymes were normal. A chest radiograph showed bilateral upper lobe patchy infiltrates with left upper lobe consolidation. Computed tomographic (CT) scan of the chest with contrast showed bilateral ground glass infiltrates with focal consolidation (Figure 1) and no evidence of pulmonary embolism. Induced sputum was negative for Pneumocystis carinii, fungi, or acid-fast bacilli. A bronchoalveolar lavage was performed. What is the diagnosis? PMID- 15336586 TI - Treatment of diabetic ketoacidosis: should mode of insulin administration dictate use of intensive care facilities? PMID- 15336587 TI - From motives to results: improving the effectiveness of quality improvement. PMID- 15336588 TI - Proposed guidelines for clenbuterol food poisoning. PMID- 15336589 TI - A new lease on life for patients with Duchenne muscular dystrophy in Japan. PMID- 15336590 TI - A hydatid cyst presenting as an axillary mass. PMID- 15336591 TI - The case for invigorating internal medicine. PMID- 15336592 TI - The biological role of strontium. AB - This review summarises old and more recent literature on the biological role of the bone-seeking trace metal strontium (Sr). It covers areas of chemistry, nutrition, toxicity, transport across biological membranes, homeostasis, general physiology, calcium-strontium interactions, and particularly the role of strontium in bone. The promoting action of strontium on calcium uptake into bone at moderate strontium supplementation, and the rachitogenic action of strontium at higher dietary strontium levels are emphasised. The literature is summarised of the novel antiosteoporotic drug strontium ranelate, which appears to act by a combination of reduced bone resorption and increased uptake of calcium into bone. PMID- 15336593 TI - Leucine7 to proline7 polymorphism in prepro-NPY gene and femoral neck bone mineral density in postmenopausal women. AB - Neuropeptide Y (NPY) is a versatile neurotransmitter that has recently been shown to regulate bone metabolism in animal and in vitro studies. We studied the influence of leucine7-to-proline7 (Leu7/Pro7) polymorphism of the NPY signal peptide gene on bone mineral density (BMD) before and after a 5-year hormone replacement therapy (HRT) in 316 early postmenopausal women participating in a randomized controlled trial nested in the population-based Kuopio Osteoporosis Risk Factor and Prevention (OSTPRE) study. The participants were randomized into two treatment groups: the HRT group (n = 146) received a sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate and calcium lactate, 500 mg/day (equal to 93 mg Ca2+) alone or in combination with vitamin D3, 100-300 IU/day. The non-HRT group (n = 170) received calcium lactate, 500 mg alone or in combination with vitamin D3, 100-300 IU/day. BMDs of the lumbar spine (L2-4) and proximal femur were measured by using dual X-ray absorptiometry (DXA). The frequency of Leu7/Pro7 polymorphism was 15.2%. At baseline, there were no significant differences in the lumbar or femoral neck BMD between the subjects who had Leu7Pro7 polymorphism and the normal subjects. After 5 years, the BMD of the femoral neck remained unaltered and that of the lumbar spine increased by 1.7% in the HRT group, whereas both BMDs were decreased by 4-5% in the non-HRT group. After 5 years, the femoral neck BMD was significantly lower in those with the wild-type NPY polymorphism than in those with Leu7/Pro7 polymorphism (P = 0.040) in the non-HRT group. In the HRT group, the changes in BMD were quite modest and not significantly modified by Leu7/Pro7 genotype. We conclude that the Leu7/Pro7 polymorphism in NPY signal gene may favorably affect femoral neck BMD in postmenopausal women. PMID- 15336594 TI - Severe osteopenia in CFTR-null mice. AB - Osteoporosis is a common complication in cystic fibrosis (CF) patients. In this study, we performed a histomorphometric analysis of the bones of a mouse genetic model of human CF in which both copies of the cystic fibrosis transmembrane conductance regulator (CFTR) gene are inactivated. We find that, even in the absence of obvious nutritional and therapeutic differences, the CFTR mutation is associated with severe osteopenia. Bone mineral density (BMD) of total body and of individual bones is significantly diminished. CFTR mutants display a striking significant (50%) reduction of cortical bone width and thinner trabeculae. Analysis of dynamic parameters indicates a significant reduction of bone formation and a concomitant strong increase in bone resorption. Active osteoclasts where found mostly associated with cortical bone. Our data support the concept that CF-associated osteoporosis is part of the syndromic symptoms associated with the CFTR mutation. PMID- 15336595 TI - Alteration of noncollagenous bone matrix proteins in distal renal tubular acidosis. AB - Our previous report on bone histomorphometry in patients with distal renal tubular acidosis (dRTA) revealed decreased bone formation rate (BFR) when compared to healthy subjects. The abnormality improved significantly after alkaline therapy. The modest increase in osteoblastic surface, after correction of metabolic acidosis, could not explain the striking improvement in bone formation, suggesting additional influence of metabolic acidosis on osteoblast function and/or bone matrix mineralization. Osteoblasts and, to a lesser extent, osteoclasts synthesize and secrete bone matrix including type I collagen and various noncollagenous proteins (NCPs). Substantial evidence suggested diverse functions of NCPs related to bone formation, resorption, and mineralization. Metabolic acidosis, through its effect on bone cells, may result in an alteration in the production of NCPs. Our study examined bone histomorphometry with detailed analysis on the mineralization parameters and NCPs expression within the bone matrix of patients with dRTA before and after treatment with alkaline. Seven dRTA patients underwent bone biopsy at their initial diagnosis and again 12 months after alkaline therapy. Bone mineral density (BMD) and bone histomorphometry were obtained at baseline and after the treatment. The expression of NCPs was examined by immunohistochemistry, quantitated by digital image analysis, and reported as a percentage of area of positive staining or mineralized trabecular bone area. Alkaline therapy normalized the low serum phosphate and PTH during acidosis. The reduction in BMD at baseline improved significantly by the treatment. Bone histomorphometry demonstrated the increase in osteoid surface and volume without significant alteration after acidosis correction. In comparison to the normal subjects, osteoid thickness was slightly but insignificantly elevated. Osteoblast and osteoclast populations and their activities were suppressed. The reduction in mineral apposition rate and adjusted apposition rate were observed in conjunction with the prolongation of mineralization lag time. Alkaline therapy improved the mineralization parameters considerably. In addition to the increase in BFR relative osteoblast number after acidosis correction, osteocalcin expression in the bone matrix increased significantly from 16.7% to 22.3%. Six of seven patients had decreased osteopontin expression. In conclusion, the abnormal bone remodeling in dRTA is characterized by low turnover bone disease with some degree of defective mineralization. Alteration of NCPs expression suggested the effect of metabolic acidosis on bone cells. Alkaline therapy increased bone mass through the restoration of bone mineral balance and, perhaps, improved osteoblast function. PMID- 15336596 TI - Effects of activated T cells on osteoclastogenesis depend on how they are activated. AB - INTRODUCTION: Activated T cells are emerging as important regulators of osteoclast function in inflammatory diseases. Both pro- and anti-resorptive properties have been described. We reasoned that this reported variability of the effects of T cells on osteoclast formation depends on how T cells are activated in vitro. METHODS: We harvested T lymphocytes from 5-week-old C57BL/6 mouse spleens. Activation was performed with anti-CD3epsilon and -CD28 Ab (Abs), concanavalin A (Con A), phytohemagglutinin (PHA), or the superantigen Staphylococcal enterotoxin A (SEA). Osteoclastogenesis was induced by receptor activator of NF-kappaB ligand (RANKL) in the mouse macrophage cell line RAW 264.7 cells, or primary macrophage CD11b+ cells from mouse spleen. Cells were cultured with T cells or with their conditioned medium. RESULTS: Co-culture of activated T lymphocytes with RAW 264.7 cells inhibited osteoclastogenesis but only when activated by Abs. This effect was CD4+ -dependent. Conditioned medium from activated T lymphocytes with Abs consistently blocked osteoclastogenesis in RAW 264.7 and CD11b+ cells. T cells activated with SEA, Con A, and PHA had inconsistent effects on osteoclastogenesis. We then tested the role of interferon (IFN)-gamma, a known inhibitor of osteoclastogenesis, in the effects of T cells on osteoclast formation. IFN-gamma neutralizing antibody blocked the inhibitory effect of T-cell conditioned medium on osteoclastogenesis. Osteoclast precursors from IFN-gamma receptor-null mice treated with 0.1% medium from activated T cells formed osteoclasts. However, higher doses of medium inhibited osteoclastogenesis, so that we cannot exclude that other factors besides IFN-gamma may be involved. CONCLUSIONS: Available methods to activate T lymphocytes result in variable effects on osteoclastogenesis. IFN-gamma is the main factor responsible for the inhibitory effects of activated T cells on osteoclast formation. PMID- 15336597 TI - Insulin-like growth factor II induces apoptosis in osteoblasts. AB - The ability of insulin-like growth factor II (IGF-II) to modulate apoptosis was studied in murine osteoblasts. At 72 h of culture, 0.01, 0.1, and 1.0 nM IGF-II produced a dose-dependent increase in apoptosis assayed by TdT-mediated dUTP biotin nick end labeling (TUNEL) and confirmed with acridine orange-ethidium bromide staining. A maximal increase of 5.0-fold above control was found with 1 nM IGF-II. A time course of treatment with 0.1 nM IGF-II demonstrated a significant increase in apoptosis compared to vehicle-treated cells by 48 h. IGF II-induced apoptosis could not be inhibited by a blocking antibody to the IGF-I receptor. Human osteoblast cultures demonstrated a similar dose-dependent increase in apoptosis with IGF-II. No significant effect of IGF-II was found on proliferation in murine osteoblast cultures. Western blot analysis demonstrated that IGF-II decreased Bcl-2 protein levels, but not Bax, resulting in a significant reduction in the Bcl-2/Bax ratio. To determine if overexpression of Bcl-2 could block IGF-II-induced apoptosis, osteoblasts were isolated from a transgenic mouse that overexpresses human Bcl-2 in bone through a construct utilizing the 2.3 kb promoter region of the Type I collagen gene linked to a 1.8 kb region of human Bcl-2 (Col2.3Bcl-2). At 72 h, IGF-II significantly increased apoptosis in a dose-dependent manner in osteoblast cultures from the control littermates. In osteoblasts from Col2.3Bcl-2 mice, no significant effect on apoptosis was found with 0.01, 0.1, or 1.0 nM IGF-II. Western blot analysis of Bcl-2 and Bax levels demonstrated a transient decrease in the Bcl-2/Bax ratio at 24 h with no decrease in the ratio at 48 or 72 h. Thus, IGF-II appears to promote osteoblast apoptosis, and overexpression of Bcl-2 is able to block IGF-II-induced apoptosis. PMID- 15336598 TI - Lipopolysaccharide stimulates expression of osteoprotegerin and receptor activator of NF-kappa B ligand in periodontal ligament fibroblasts through the induction of interleukin-1 beta and tumor necrosis factor-alpha. AB - Our recent work showed that human periodontal ligament fibroblasts (HPLF) secrete bioactive osteoprotegerin (OPG), which inhibits osteoclastic differentiation and activity. However, it is unknown how HPLF regulate bone metabolism in the presence of lipopolysaccharide (LPS), which is a cell component of gram-negative bacteria and a pathogen in inflammatory bone diseases such as periodontitis. The present study examined the effects of Escherichia coli LPS on the gene expression of interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), OPG, and receptor activator of NF-kappa B ligand (RANKL) in HPLF using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis. In HPLF cultured with LPS for 48 h, expression of both OPG and RANKL mRNA was up-regulated, whereas for up to 24 h of stimulation, such up-regulation was not observed. However, LPS increased expression of IL-1 beta and TNF-alpha mRNA within 6 h of treatment. Moreover, in HPLF cultured with IL-1 beta or TNF alpha, OPG and RANKL expression was induced within 12 h of culture. The administration of neutralizing antibodies against human IL-1 beta or TNF-alpha to LPS-treated cultures of HPLF inhibited the induction of OPG and RANKL expression. These suggest that LPS stimulates both OPG and RANKL expression in HPLF by up regulating IL-1 beta and TNF-alpha. In addition, administration of conditioned medium (CM) from HPLF (HPLF-CM) stimulated with LPS for 48 h to mouse bone marrow culture failed to induce osteoclast-like cell (OCL) formation. When mouse spleen cells were cocultured with HPLF in the presence of LPS, OCL formation was completely blocked. Taken together, our results indicate that human periodontal ligament cells stimulated with LPS inhibit osteoclastogenesis by producing more effective OPG than RANKL via the induction of IL-1 beta and TNF-alpha. PMID- 15336599 TI - The flurbiprofen derivatives HCT1026 and HCT1027 inhibit bone resorption by a mechanism independent of COX inhibition and nitric oxide production. AB - Prostaglandins and nitric oxide both modulate bone resorption and bone formation. We previously reported that a nitrosylated derivative of flurbiprofen, termed HCT1026, exerted inhibitory effects on osteoclastic bone resorption, which could not be reproduced by combining the parent compound with nitric oxide (NO) donors. The aim of this study was to investigate the mechanism by which HCT1026 inhibits bone resorption. We compared the effects of flurbiprofen and HCT1026 on osteoclast and osteoblast activity with those of HCT1027--an analogue of HCT1026, which lacks an NO-donating moiety. We found that HCT1026 and HCT1027 inhibited bone resorption in interleukin (IL)-1-stimulated murine osteoblast-bone marrow cocultures, with half-maximal effects (IC50) at 20 +/- 5 microM for HCT1026 and 25 +/- 6 microM for HCT1027 compared with 399 +/- 25 microM for flurbiprofen (P < 0.0001). These differences were unrelated to cyclooxygenase (COX) inhibition since HCT1026 and HCT1027 were about seven to eight times less potent than flurbiprofen at inhibiting COX-1 activity and half as potent at inhibiting COX-2 activity. Further studies showed that HCT1026 and HCT1027 activated caspase-3 in rabbit osteoclasts and promoted osteoclast apoptosis, as assessed by nuclear morphology and TUNEL assays. We conclude that HCT1026 and HCT1027 inhibit osteoclast formation and activity by a mechanism that is independent of NO production and COX inhibition. This raises the possibility that both compounds interact with a novel molecular target expressed on osteoclasts to promote apoptosis and inhibit bone resorption. This demonstrates that HCT1026 and derivatives could represent a novel class of antiresorptive drugs with therapeutic value in the treatment of bone diseases associated with accelerated bone loss due to osteoclast activation. PMID- 15336600 TI - Intensity-related differences in collagen post-translational modification in MC3T3-E1 osteoblasts after exposure to low- and high-intensity pulsed ultrasound. AB - Low-intensity pulsed ultrasound (LIPUS) has distinct effects on biologic mineralization at intensities of <100 mW/cm2. Intensity-dependent differences in the pattern of accelerated mineralization may be due to different alterations in regulation of collagenous matrix formation. However, little is known about the influence of LIPUS on collagen metabolism in the context of mineralization processes. Therefore, we attempted to evaluate differential effects of two intensities of pulsed ultrasound (30 vs. 120 mW/cm2) on collagen post translational modification and mineralization in osteoblastic MC3T3-E1 cells. Murine osteoblastic MC3T3-E1 cells were exposed to pulsed ultrasound (1.5-MHz, 200-ms burst sine wave at 1.0-kHz frequency, either 30 or 120 mW/cm2 SATA, for 20 min/day from Day 14 to Day 35 postconfluence). Expression patterns of lysyl oxidase (LO), procollagen-lysine, 2-oxyglutarate, 5-dioxigenase 1 (PLOD1, LH1), and 2 (PLOD2, LH2) was examined using quantitative PCR. Quantitative analysis of reducible immature cross-links (dihydroxylysinonorleucine, hydroxylysinonorleucine, and lysinonorleucine) and nonreducible mature cross links (pyridinoline and deoxypyridinoline) as well as analysis of the maturation of immature to mature cross-links were performed. Exposure to 30 mW/cm2 LIPUS upregulated LH2 mRNA expression and enzyme activity compared to controls. It was associated with increased relative amounts of telopeptidyl hydroxylysine (Hyl) derived cross-links beginning on Day 14, upregulated LO mRNA expression, increased total reducible and nonreducible cross-links, and increased ratios of newly formed nonreducible to reducible cross-links. Similarities in the pattern of cross-link formation and calcium deposition in matrices between 30 mW/cm2 LIPUS-treated MC3T3-E1 cultures and bone suggest that 30 mW/cm2 LIPUS may promote the maturation of collagenous matrix as a scaffold for calcification. In contrast, exposure to 120 mW/cm2 ultrasound increased calcium accumulation compared to control at Day 35, but increases were delayed until Day 25. No differences in the extent and pattern of cross-links were observed compared to controls. These results suggest that the promotion of mineralization induced by 120 mW/cm2 may be attributed to other factors involved in mineralization process rather than cross-link pattern. Our results demonstrated the existence of differential effects of lower versus higher intensities of ultrasound on mineralization processes in vitro. PMID- 15336601 TI - Annexin V disruption impairs mechanically induced calcium signaling in osteoblastic cells. AB - The mechanical environment of the skeleton plays an important role in the establishment and maintenance of structurally competent bone. Biophysical signals induced by mechanical loading elicit a variety of cellular responses in bone cells, however, little is known about the underlying mechanotransduction mechanism. We hypothesized that bone cells detect and transduce biophysical signals into biological responses via a mechanism requiring annexin V (AnxV). AnxV, a calcium-dependent phospholipid binding protein, has several attributes, which suggest it is ideally suited for a role as a mechanosensor, possibly a mechanosensitive ion channel. These include the ability to function as a Ca2+ selective ion channel, and the ability to interact with both extracellular matrix proteins and cytoskeletal elements. To test the hypothesis that AnxV has a role in mechanosensing, we studied the response of osteoblastic cells to oscillating fluid flow, a physiologically relevant physical signal in bone, in the presence and absence of AnxV inhibitors. In addition, we investigated the effects of oscillating flow on the cellular location of AnxV. Oscillating fluid flow increased both [Ca2+]i levels and c-fos protein levels in osteoblasts. Disruption of AnxV with blocking antibodies or a pharmacological inhibitor, K201 (JTV-519), significantly inhibited both responses. Additionally, our data show that the cellular location of AnxV was modulated by oscillating fluid flow. Exposure to oscillating fluid flow resulted in a significant increase in AnxV at both the cell and nuclear membranes. In summary, our data suggest that AnxV mediates flow induced Ca2+ signaling in osteoblastic cells. These data support the idea of AnxV as a Ca2+ channel, or a component of the signaling pathway, in the mechanism by which mechanical signals are transduced into cellular responses in the osteoblast. Furthermore, the presence of a highly mobile pool of AnxV may provide cells with a powerful mechanism by which cellular responses to mechanical loading might be amplified and regulated. PMID- 15336602 TI - Calcium-sensing receptor activation stimulates parathyroid hormone-related protein secretion in prostate cancer cells: role of epidermal growth factor receptor transactivation. AB - We have previously reported that high extracellular Ca2+ stimulates parathyroid hormone-related protein (PTHrP) release from human prostate and breast cancer cell lines as well as from H-500 rat Leydig cancer cells, an action mediated by the calcium-sensing receptor (CaR). Activating the CaR leads to phosphorylation of mitogen-activated protein kinases (MAPKs) that participate in PTHrP synthesis and secretion. Because the CaR is a G protein-coupled receptor (GPCR), it is likely to transactivate the epidermal growth factor receptor (EGFR) or the platelet-derived growth factor receptor (PDGFR). In this study, we hypothesized that activation of the CaR transactivates the EGFR or PDGFR, and examined whether transactivation affects PTHrP secretion in PC-3 human prostate cancer cells. Using Western analysis, we observed that an increase in extracellular Ca2+ resulted in delayed activation of extracellular signal-regulated kinase (ERK) in PC-3 cells. Pre-incubation with AG1478 (an EGFR kinase inhibitor) or an EGFR neutralizing antibody inhibited the high Ca2+ -induced phosphorylation of ERK1/2. GM6001, a pan matrix metalloproteinase (MMP) inhibitor, also partially suppressed the ERK activation, but AG1296 (a PDGFR kinase inhibitor) did not. High extracellular Ca2+ stimulates PTHrP release during a 6-h incubation (1.5- to 2.5- and 3- to 4-fold increases in 3.0 and 7.5 mM Ca2+, respectively). When cells were preincubated with AG1478, GM6001, or an antihuman heparin-binding EGF (HB-EGF) antibody, PTHrP secretion was significantly inhibited under basal as well as high Ca2+ conditions, while AG1296 had no effect on PTHrP secretion. Taken together, these findings indicate that activation of the CaR transactivates the EGFR, but not the PDGFR, leading to phosphorylation of ERK1/2 and resultant PTHrP secretion, although CaR-EGFR-ERK might not be the only signaling pathway for PTHrP secretion. This transactivation is most likely mediated by activation of MMP and cleavage of proheparin-binding EGF (proHB-EGF) to HB-EGF. PMID- 15336603 TI - Glucocorticoid regulation of human BMP-6 transcription. AB - Addition of dexamethasone (Dex) to human mesenchymal stem cells (hMSCs) resulted in a 16-fold increase in human bone morphogenetic protein-6 (hBMP-6) mRNA levels 24 h after treatment. Evaluation of luciferase expression after transfection of HeLa cells with hBMP-6 promoter/luciferase reporter constructs indicated that the hBMP-6 promoter activity was contained in a 268-bp region (-1051 to -784 where +1 is the translation start site) over 600 bases 5' to that previously published. It further showed that the promoter activity is regulated by glucocorticoid treatment. Analysis of RNA from hMSCs and HeLa cells by primer extension, RNase protection, and 5' RACE further narrowed the location of the transcription start site to an 84-bp region (-940 to -857). To determine whether this start site was regulated in hMSCs, hBMP-6 mRNA levels in control and Dex-treated cells were quantitated by RT-PCR using one primer set in the translated region of the gene and one located just 3' of the 84-bp region. Both primer sets showed hBMP-6 mRNA levels approximately 16- to 22-fold higher in the Dex-treated cells, demonstrating that hBMP-6 transcription is being regulated by glucocorticoids in the pluripotent hMSCs at the upstream transcription start site. PMID- 15336604 TI - The oxidative metabolism of estrogen modulates response to ERT/HRT in postmenopausal women. AB - We have previously demonstrated that estrogen metabolism is one of the determinants of bone density after menopause. Increased hydroxylation to relatively nonestrogenic metabolites 2-hydroxyestrone (2OHE1) and 2 methoxyestrone (2MeOE1) was associated with low bone mineral density (BMD), while increased hydroxylation to the potent 16alpha-hydroxyestrone (16alphaOHE1) and weakly estrogenic estriol (E3) was associated with higher BMD. In this study, we tested the hypothesis that response to estrogen-hormone replacement therapy (ERT/HRT) is also related to individual differences in estrogen metabolism. Urinary estrogen metabolites were measured in 310 postmenopausal women using ESTRAMET enzyme immunoassay kit. Of these, 163 were on HRT with conjugated equine estrogen (CEE) and medroxyprogesterone acetate (MPA, Premarin and Provera) or ERT with conjugated equine estrogen alone (Premarin), and 147 women not on ERT/HRT acted as comparison. Annual rates of BMD changes were calculated on a subset of 81 women on ERT/HRT who had more than one previous BMD measured by dual-energy X ray absorptiometry (DEXA). Controlling for age, years since menopause (YSM), body mass index (BMI), waist to hip ratio, and smoking, we found that urinary estrogen metabolite levels were significantly higher in ERT/HRT-treated women compared to those not on ERT/HRT. Furthermore, women in the higher 2 tertiles of 2OHE1 and 2OHE1/16aOHE1 ratio had positive increments in BMD compared to those in the lowest tertile who lost bone while on ERT/HRT. Thus, women with estrogen metabolism favoring the 2-hydroxylation pathway respond favorably to ERT/HRT. PMID- 15336605 TI - Family 2 cystatins inhibit osteoclast-mediated bone resorption in calvarial bone explants. AB - Osteoclastic bone resorption depends on the activity of various proteolytic enzymes, in particular those belonging to the group of cysteine proteinases. Biochemical studies have shown that cystatins, naturally occurring inhibitors of these enzymes, inhibit bone matrix degradation. Since the mechanism by which cystatins exert this inhibitory effect is not completely resolved yet, we studied the effect of cystatins on bone resorption microscopically and by Ca-release measurements. Calvarial bone explants were cultured in the presence or absence of family 2 cystatins and processed for light and electron microscopic analysis, and the culture media were analyzed for calcium release. Both egg white cystatin and human cystatin C decreased calcium release into the medium significantly. Microscopic analyses of the bone explants demonstrated that in the presence of either inhibitor, a high percentage of osteoclasts was associated with demineralized non-degraded bone matrix. Following a 24-h incubation in the presence of cystatin C, 41% of the cells were adjacent to areas of demineralized non-degraded bone matrix, whereas in controls, this was only 6%. If bone explants were cultured with both PTH and cystatin C, 60% of the osteoclasts were associated with demineralized non-degraded bone matrix, compared to 27% for bones treated with PTH only (P < 0.01). Our study provides evidence that cystatins, the naturally occurring inhibitors of cysteine proteinases, reversibly inhibit bone matrix degradation in the resorption lacunae adjacent to osteoclasts. These findings suggest the involvement of cystatins in the modulation of osteoclastic bone degradation. PMID- 15336606 TI - Administration of estrogen shortly after ovariectomy mimics the anti-nociceptive action and change in 5-HT1A-like receptor expression induced by calcitonin in ovariectomized rats. AB - Although many clinical reviews are consistent with the view that hormone replacement therapy should be recommended for increasing bone mass of osteoporotic patients, calcitonin administration is preferable to hormone replacement therapy for the alleviation of pain accompanying osteoporosis, despite the fact that osteoporosis and the accompanying pain are accelerated by the reduction in estrogen levels. Distinct from the clinical view, animal studies have shown that estrogen treatment reduces ovariectomy-induced hyperalgesia, although the mechanism of this phenomenon is unknown. The discrepancy in clinical and animal study outcomes may be due to the timing of administration of estrogen after depletion of the hormone. To address this possibility, the anti-nociceptive effect of estrogen was compared with calcitonin using the tail withdrawal test in rats injected with estrogen or calcitonin at 3 weeks (short term) or 15 weeks (long term) after ovariectomy. Furthermore, we analyzed the change in [3H]8-OH DPAT binding in the spinal cord, addressing whether estrogen exerts its anti nociceptive effect by the expression of 5-HT receptors attributable to calcitonin induced analgesia, as has been reported in our previous animal studies. The present study demonstrates that the administration of estrogen injected in the short term, but not long term, after ovariectomy mimicked the effects of calcitonin-induced anti-nociception and prevention of ovariectomy-induced decrease in 5-HT receptor expression in the spinal cord, although the effects of calcitonin were observed regardless of the timing of calcitonin injection. These results suggest that the estrogen receptor is downregulated gradually after ovariectomy. Disappearance of the estrogen receptor may be one of the reasons that estrogen is not recommended for the treatment for chronic pain associated with osteoporosis. PMID- 15336607 TI - 1 Alpha-hydroxyvitamin D2 and 1 alpha-hydroxyvitamin D3 have anabolic effects on cortical bone, but induce intracortical remodeling at toxic doses in ovariectomized rats. AB - It is well established that vitamin D metabolites have anabolic properties on cancellous bone in rats. However, few data are available on cortical bone effects of vitamin D metabolites. In this study, we examined the effects of the synthetic vitamin D analogs 1alpha-hydroxyvitamin D2 (1alpha(OH)D2) and 1alpha hydroxyvitamin D3 (1alpha(OH)D3) on cortical bone of the tibial shaft in ovariectomized (OVX) rats using bone histomorphometry. Six-month-old Fischer 344 rats were either OVX or sham-operated (SHAM). OVX rats received vehicle, 1alpha(OH)D2 or 1alpha(OH)D3 orally via the diet in a dose range from 0.025 to 0.2 microg/kg/day. All animals were killed 3 months postsurgery after in vivo fluorochrome labeling. Relative to SHAM rats, vehicle-treated OVX rats showed a reduction in cortical bone area (%) due to expansion of the marrow cavity. Treatment of OVX rats with either 1alpha(OH)D2 or 1alpha(OH)D3 dose-dependently decreased marrow area, and increased cortical area, periosteal perimeter, and periosteal and endocortical bone formation rate compared with OVX vehicle controls. Interestingly, OVX animals receiving the highest doses showed intracortical resorption cavities, a phenomenon only exceptionally observed in rats. The intracortical hole area was significantly lower in 1alpha(OH)D2-treated compared with 1alpha(OH)D3-treated rats. We conclude that 1alpha(OH)D2 and 1alpha(OH)D3 prevent cortical bone loss in OVX rats and have anabolic effects on cortical bone at higher doses. However, very high, toxic doses of both vitamin D analogs induce intracortical remodeling as an untoward side effect. PMID- 15336608 TI - In vivo and in vitro evidence that the high osteoblastic activity in C3H/HeJ mice compared to C57BL/6J mice is intrinsic to bone cells. AB - Two inbred mouse strains, C3H/HeJ (C3H) and C57BL/6J (B6), displayed a profound difference in femoral peak bone density. We have previously shown that the difference could be attributed to a greater bone formation rate (BFR) that was due to a higher osteoblastic activity [measured by a mineral apposition rate (MAR)] in the C3H (high density) than B6 (low density) mice. The present study sought to determine (1) whether the BFR/MAR differences between the two mouse strains present in weight-loaded endochondral bones are also seen in less weight loaded membranous bones and (2) whether the difference in osteoblastic activity was seen in vitro in the absence of systemic factors. To address the first objective, we performed histomorphometric measurements on the weakly loaded membranous bones (i.e., parietal bones of the calvaria) to determine if there were similar differences in MAR and BFR of membranous bones as those of highly loaded, endochondral bones. The parietal bones of adult C3H mice showed similar increases in MAR and BFR as the endochondral bones, compared to B6 mice of same age, suggesting that the differences in the MAR and BFR in the two mouse strains are probably not related to differences in mechanical strain. These findings also suggest that the gene(s) responsible for the difference in MAR between strains may not be a mechanical response gene. With respect to the second objective, we isolated osteoblasts from the parietal bones and determined their differentiation status (i.e., ALP-specific activity) and bone-forming ability (i.e., mineralized nodule formation) in vitro. Consistent with the premise that C3H osteoblasts have an intrinsic, higher differentiation status and bone-forming ability than B6 osteoblasts, osteoblasts isolated from C3H mice as compared with those from B6 mice had a significantly greater ALP-specific activity and a greater ability to form mineralized nodules in vitro in the absence of systemic factors. Because differences in ALP activity, bone-forming ability, cortical bone width, and osteoblastic activity were detected at birth, the different MAR/BFR phenotypes develop at very early life and even perhaps during embryogenesis. In conclusion, we have for the first time provided evidence that the genetic differences responsible for the observed MAR/BFR phenotype in the C3H-B6 strains are intrinsic to osteoblasts and might not depend on responses to mechanical loading and/or alterations in systemic factors. PMID- 15336609 TI - Spinal maturation affects vertebral compressive mechanics and vBMD with sex dependence. AB - The effects of natural aging on the mechanics of the spine are far better understood for the mature adult spine than for the developing (immature) spine. Throughout its chondrification and ossification, the vertebra, which is the primary structural unit of the spine, undergoes enormous cellular, biochemical, and structural changes that should strongly influence its biomechanical response to external forces. Unfortunately, very little data exist for the mechanics of immature vertebrae. Vertebral maturation was therefore investigated in 22 baboon thoracic specimens to elucidate its relationship with biomechanics and volumetric bone mineral density (vBMD). Cadaveric baboon vertebrae were used due to the limited availability of human tissues in the pediatric age range. The specimen ages ranged between 1 and 30 human-equivalent years based on skeletal maturity. Isolated ninth thoracic vertebrae (T9) were subjected to compressive loading to document their compressive mechanical properties (yield load, stiffness, yield strength, and elastic modulus) and ashed to determine their volumetric bone mineral density. Spinal maturation was discovered to significantly increase vBMD (P < 0.0001) and compressive mechanics (stiffness, bulk elastic modulus, failure load, and bulk strength, P < 0.001) in a sex-dependent manner. Vertebral stiffness increased from 1218 N/mm at 1 year to 3534 N/mm at 30 years with a second order polynomial "maturation" relationship. Volumetric bone mineral density and vertebral cross-sectional area together described the developmental patterns of stiffness and yield load of isolated vertebrae. Sex differences were observed throughout development, demonstrating differing growth patterns to accommodate mechanical loading whereby males develop larger size vertebrae and females achieve their mechanical stiffness and strength through greater bone mineral density. PMID- 15336610 TI - Determinants of ovine compact bone viscoelastic properties: effects of architecture, mineralization, and remodeling. AB - Significant decreases in ovine compact bone viscoelastic properties (specifically, stress-rate sensitivity, and damping efficiency) are associated with three years of ovariectomy and are particularly evident at higher frequencies [Proc. Orthop. Res. Soc. 27 (2002) 89]. It is unclear what materials or architectural features of bone are responsible for either the viscoelastic properties themselves, or for the changes in those properties that were observed with estrogen depletion. In this study, we examined the relationship between these viscoelastic mechanical properties and features involving bone architecture (BV/TV), materials parameters (ash density, %mineralization), and histologic evidence of remodeling (%remodeled, cement line interface). The extent of mineralization was inversely proportional to the material's efficiency in damping stress oscillations. The damping characteristics of bone material from ovariectomized animals were significantly more sensitive to variation in mineralization than was bone from control animals. At low frequencies (6 Hz or less), increased histologic evidence of remodeling was positively correlated with increased damping efficiency. However, the dramatic decreases in stress-rate sensitivity that accompanied 3-year ovariectomy were seen throughout the bone structure and occurred even in areas with little or no secondary Haversian remodeling as well as in areas of complete remodeling. Taken together, these data suggest that, while the mineral component may modify the viscoelastic behavior of bone, the basic mechanism underlying bone viscoelastic behavior, and of the changes in that behavior with estrogen depletion, reside in a non-mineral component of the bone that can be significantly altered in the absence of secondary remodeling. PMID- 15336611 TI - Effects of acute 5-fluorouracil chemotherapy and insulin-like growth factor-I pretreatment on growth plate cartilage and metaphyseal bone in rats. AB - With the intensified use of chemotherapy and improved survival rates for childhood malignancies, it has become increasingly apparent that some children or adult survivors show poor bone growth and develop osteoporosis. As a step to investigate underlying mechanisms, this project examined short-term effects in rats of chemotherapy agent 5-fluorouracil (5-FU) on cell proliferation, apoptosis, and bone formation at tibial growth plate cartilage and its adjacent bone-forming region metaphysis. In addition, since insulin-like growth factor (IGF-I) is important for bone growth, we examined whether IGF-I pretreatment would potentially protect growth plate cartilage and bone cells from chemotherapy damage. Two days after a single high dose of 5-FU injection, proliferation of growth plate chondrocytes and metaphyseal osteoblasts/preosteoblasts was dramatically suppressed, and apoptosis was induced among osteoblasts and preosteoblasts. As a result, there was a reduction in the chondrocyte number and zonal height at the proliferative zone and a decline in the number of osteoblasts and preosteoblasts on the metaphyseal trabecular bone surface. At day 2, no obvious deleterious effects were observed on the height of the growth plate hypertrophic zone and the bone volume fraction of the metaphyseal primary spongiosa trabeculae. At day 10, while cell proliferation and growth plate structure returned to normal, there were slight decreases in trabecular bone volume, body length increase, and tibial length. While pretreatment with 1-week IGF-I systemic infusion did not attenuate the suppressive effect of 5-FU on proliferation in both growth plate and metaphysis, it significantly diminished apoptotic induction in metaphysis. These results indicate that growth plate cartilage chondrocytes and metaphyseal bone cells are sensitive to chemotherapy drug 5-FU and that IGF-I pretreatment has some anti-apoptotic protective effects on metaphyseal bone osteoblasts and preosteoblasts. PMID- 15336612 TI - Femoral bone structural geometry adapts to mechanical loading and is influenced by sex steroids: the Penn State Young Women's Health Study. AB - We used 10 years of longitudinal data from Penn State Young Women's Health Study to explore predictors of adult bone structural geometry and strength. One hundred twelve participants were enrolled in the study at age 12. We report findings on the 76 participants who remained in the study for 10 years. Measurements were recorded biannually for the first 4 years and annually thereafter. Proximal femur DXA scans (Hologic QDR 2000) were taken from 17-22 years and analyzed using a hip structure analysis program to assess areal bone mineral density (BMD, g/cm2), subperiosteal width, cortical thickness, bone cross-sectional area (CSA), and section modulus (Z) at the narrow neck and femoral shaft. Total body lean mass (g) was measured with DXA total body scans. Nutrition, anthropometry, and sex steroids [testosterone (T) and estradiol (E2)] were measured from ages 12-22 years. Multiple regression models were used to assess predictors of change in bone variables (17-22 years) and absolute bone values (average of age 21 and 22 years, n = 79). Neck Z (+3.1%) and width (+1.3%), but not BMD (-0.8%), increased significantly from age 17 to 22 years. At the shaft, all variables increased (+1.0-4.0%, P < 0.01). After controlling for baseline (age 17) height, weight and bone measurement, weight change (neck) or lean mass (shaft), and age of menarche were the primary predictors of change in bone strength. After controlling for height and weight, only lean mass predicted absolute young adult Z at both the neck (r2 = 0.48, P < 0.01) and the shaft (r2 = 0.67, P < 0.01). When lean mass was removed from the model, sports exercise score replaced lean mass as a predictor of Z at both neck (r2 = 0.40, P < 0.01) and shaft (r2 = 0.60, P < 0.01) sites. For neck and shaft cortical thickness and BMD, both estradiol and sports score/lean mass were positive predictors (r2 = 0.15-0.40, P < 0.01). For neck bone width, testosterone levels (negative) and lean mass (positive) were significant (r2 = 0.48). Results were similar for each geometric variable at the shaft site. These data suggest that bone adapts its bending strength primarily to mechanical loading (represented by lean mass and sports exercise score) and that sex steroids are associated with bone geometric structure. PMID- 15336613 TI - Association of plasma folate, plasma total homocysteine, but not methylenetetrahydrofolate reductase C667T polymorphism, with bone mineral density in postmenopausal Iranian women: a cross-sectional study. AB - Polymorphisms of methylenetetrahydrofolate reductase (MTHFR) have been well documented to cause hyperhomocysteinemia, and recent studies suggest an association of C677T mutation of methylenetetrahydrofolate reductase with low bone mineral density (BMD). In this study, the association of plasma total homocysteine (Hcy), plasma folate, and vitamin B12 as well as methylenetetrahydrofolate reductase C667T polymorphism with bone mineral density at neck of femur and lumbar spine in 271 postmenopausal Iranian women was investigated. Bone mineral density was measured by dual-energy X-ray absorptiometry. Restriction fragment length polymorphism was used for genotyping the methylenetetrahydrofolate reductase polymorphism. Plasma total homocysteine, plasma folate, and vitamin B12 were also determined. The bone mineral densities at the neck of femur and lumbar spine together with other clinical characteristics among methylenetetrahydrofolate reductase genotypes (CC, CT, and TT) were examined. Bone mineral densities at both neck of femur (r = -0.18, P = 0.003) and lumbar spine (r = -0.16, P = 0.01) were significantly and negatively correlated with the logarithm of plasma total homocysteine. Bone mineral density at the lumbar spine was also significantly and positively associated with plasma folate (r = 0.14, P = 0.02). However, no correlation between methylenetetrahydrofolate reductase polymorphism with bone mineral density at neck of femur (r = -0.01, P = 0.81) and lumbar spine (r = -0.04, P = 0.51) was observed. The negative association of plasma total homocysteine with bone mineral density was no longer significant when adjusted for folate and vitamin B12. Plasma folate and age were the main predictors of plasma total homocysteine explaining 15.3% and 5.2% of the variance of plasma total homocysteine, respectively. Methylenetetrahydrofolate reductase polymorphism, however, was not associated with plasma folate (r = 0.086, P = 0.17) or vitamin B12 (r = 0.05, P = 0.4). Plasma folate was one of the main predictors explaining 3.0% and 1.7% of variance of the bone mineral density at femoral neck and lumbar spine, respectively. Results from this study suggest hyperhomocysteinemia as a result of folate deficiency, but not methylenetetrahydrofolate reductase polymorphism, is independently associated with low bone mineral density and may contribute to the pathogenicity of osteoporosis in postmenopausal Iranian women. PMID- 15336614 TI - The effect of atorvastatin on markers of bone turnover in patients with type 2 diabetes. AB - Retrospective and uncontrolled studies suggest that the lipid-lowering statin class of drugs has either no or beneficial effects on bone density and may reduce fracture risk. We have examined the effects of atorvastatin on serum and plasma markers of bone turnover in 25 patients (age 56 +/- 8 years) with type 2 diabetes (duration: 4.7 +/- 5.0 years, 16 female, 2 insulin-treated, 4 diet alone, and 19 on oral hypoglycemic agents) and baseline hypercholesterolemia (cholesterol 6.6 +/- 0.8 mmol/l) in a double-blind, placebo-controlled, crossover study of 12 weeks of placebo/40 mg of atorvastatin with an 8-week wash-out period. Atorvastatin resulted in a fall in total cholesterol of 2.3 +/- 0.9 mmol/l. There were no effects of active or placebo therapy on total alkaline phosphatase, bone specific alkaline phosphatase, osteocalcin, or beta C-telopeptide of type 1 collagen (beta-CTX). We conclude that atorvastatin (40 mg/day) has no significant effect on bone turnover in man. PMID- 15336615 TI - Prevalence and correlates of vertebral fractures in adults with cystic fibrosis. AB - INTRODUCTION: Osteoporosis associated with cystic fibrosis (CF) is becoming increasingly important as the life expectancy of patients continues to increase. MATERIALS AND METHODS: We studied 191 adults with CF (18-50 years old; 100 men, 91 women). Total body, lumbar spine, and total proximal femur bone mineral density (BMD) were measured by dual-energy X-ray absorptiometry, and lateral spinal radiographs were taken for assessment of vertebral fractures. A range of anthropometric, clinical and biochemical variables were evaluated as potential correlates. RESULTS: BMD T score values at the lumbar spine lower than -2.5 SD were observed in 27.3% and 11.2% of male and female patients, respectively. These proportions fell to 14% and 9.9% for total hip and 10.4% and 12.1% for total body, in men and women, respectively. Vertebral deformities were identified in 26.7% of the patients with a slightly higher prevalence in males (32%) than in females (21%, P = 0.058). Multiple vertebral deformities were observed in 12% and 7.7% of men and women, respectively. BMD values were significantly related to body weight, FEV1, age of puberty and occasionally to cumulative steroid dose in both genders. BMD values were also significantly related with serum albumin, IgG and cholinesterase. Serum estradiol levels were found below the normal range in 23% of the women and 27% of the men, and was significantly related to femur BMD values in both women and men. Significantly lower serum estradiol and free testosterone levels were observed in men with vertebral fractures. Serum osteocalcin was below the normal range in 36% and urinary deoxypyridinoline above the normal range in 51% of the patients. CONCLUSIONS: This study indicates that osteoporosis is a common complication of CF, being related to disease progression and apparently due to both excess bone resorption and inadequate bone formation. Estradiol deficiency may have a significant role in the pathogenesis in both genders. Vertebral fracture prevalence is high and greater than expected from prevalent BMD values. PMID- 15336617 TI - High serum osteoprotegerin levels in patients with hyperthyroidism: effect of medical treatment. AB - This study was aimed at evaluating serum osteoprotegerin (OPG) concentrations in a cohort of patients with hyperthyroidism before and after methimazole (MMI) treatment. One hundred fourteen hyperthyroid patients [93 with Graves disease (GD) and 21 with toxic nodular goitre (TNG)] and 68 matched for sex and age healthy subjects were evaluated for serum free-thyroxine (FT4), free triiodiothyronine (FT3), thyrotropin (TSH), TSH receptor antibodies (TRAb), bone alkaline phosphatase (BALP), C-telopeptides of type-1 collagen (CrossLaps), OPG levels, and bone mineral density (BMD). In hyperthyroid patients, the biochemical evaluations were performed before and after 6 and 12 months of MMI treatment, whereas BMD was measured at baseline and after 12 months of treatment. Hyperthyroidism was more severe in GD than TNG patients. Serum OPG levels were found to be significantly higher in hyperthyroid patients than in the healthy subjects (4.3 pmol/l, range: 1.6-12.0, vs. 2.2 pmol/l, range: 1.4-6.0; P < 0.001), the values being higher in GD patients than TNG. A significant correlation between serum OPG levels and age was found in the healthy subjects (r: 0.48; P < 0.001) but not in hyperthyroid patients (r: -0.03; P = 0.8). In the healthy subjects, serum OPG levels were also positively correlated with both serum FT4 (r: 0.23; P = 0.03) and FT3 (r: 0.24; P = 0.04) levels. In hyperthyroid patients, however, serum OPG was still correlated with FT3 levels (r: 0.38; P < 0.001), whereas the correlation with serum FT4 was lost (r: 0.19; P = 0.06). In hyperthyroid patients, but not in the healthy subjects, serum OPG levels were correlated positively with CrossLaps (r: 0.20; P = 0.03) and negatively with BALP (r: -0.24; P = 0.01) and BMD (r: -0.33; P = 0.01). After 6 months of MMI treatment, serum OPG concentrations decreased significantly in TNG patients (from 3.5 pmol/l, range: 1.6-8.0, to 2.3 pmol/l, range: 1.0-4.3; P < 0.001), whereas a not significant change in OPG levels occurred in GD patients (from 4.8 pmol/l, range: 1.8-12.0, to 4.2 pmol/l, range: 1.0-14.0; P = 0.7). At Month 12 of treatment, serum OPG concentrations were significantly lower than those measured at baseline in both TNG (2.5 pmol/l, range: 1.0-3.1, vs. 3.5 pmol/l, range: 1.6 8.0; P < 0.001) and GD (2.1 pmol/l, range: 1.0-8.6, vs. 4.8 pmol/l, range: 1.8 12.0; P < 0.001). At this time, no significant differences in serum OPG, CrossLaps, and BALP values were found between patients and control subjects. At the end of follow-up, BMD was higher than those measured at baseline but still significantly lower than those measured in the control subjects. This study shows that hyperthyroid patients have serum OPG concentrations significantly higher in comparison with euthyroid subjects, in relation to thyroid hormone excess and high bone turnover. Medical treatment of hyperthyroidism normalizes serum OPG levels in temporal relationship with the normalization of bone metabolism markers, even in presence of persistent abnormal bone structure as determined by ultrasonography. PMID- 15336616 TI - A follow-up linkage study for bone size variation in an extended sample. AB - Bone size, which has strong genetic determination, is an important determinant of bone strength and a risk factor of osteoporotic fractures. We previously reported an approximately 10-cm genome-wide linkage scan in 630 subjects from 53 US Caucasian pedigrees. The strongest evidence of linkage was obtained on chromosome 17q22 near the marker D17S787, with a two-point LOD score of 3.98 and a multipoint maximum LOD score (MLS) of 3.01. Additionally, suggestive linkages (1.54 < MLS < 2.83) were found at the other four chromosomal regions. In the present study, with an attempt to further examine our previous findings, we perform a follow-up linkage analysis in an expanded sample of 79 pedigrees with 1816 subjects. The total sample contains >80,000 informative relative pairs for linkage analyses, including 3846 sib pairs. Fifteen markers covering the above five promising regions are genotyped, narrowing the average genomic distance from approximately 10 to 5 cm. In the total 79 pedigrees, support of linkage was achieved for the wrist bone size at 17q22 with a two-point LOD score of 2.27 (P = 0.0006) and MLS of 1.78 (P = 0.002). The genomic region 17q22 includes COL1A1, a strong candidate gene that is significantly associated with osteoporotic fracture risk. Our data suggest that this region is promising for further exploratory studies. PMID- 15336618 TI - Body composition changes with age have gender-specific impacts on bone mineral density. AB - Body weight, smoking, alcohol, physical activity, and diet have been proven to affect bone mineral density (BMD) directly or indirectly. Of these, body weight is perhaps best known to affect BMD. However, there is some debate as to whether lean body mass (LBM) or fat mass (FM), the two components of body weight, most determines BMD. Recently, newer peripheral densitometry devices have been developed, which have the advantages of low cost and portability, and this has made field epidemiologic study of osteoporosis possible. As the number of studies that have focused on the contribution made by body composition to BMD is limited, we investigated the relative contribution of LBM and FM to BMD in healthy Korean subjects. 402 age- and weight-matched subjects over 45 years old were selected from a population-based cohort. The mean ages of men and women were 64.1 +/- 8.7 (mean +/- SD) and 64.2 +/- 12.7 years, and mean weights were 63.0 +/- 8.2 and 63.1 +/- 8.2 kg, respectively. BMD was measured by peripheral dual-energy X-ray absorptiometry (DEXA) and body composition by bioelectrical impedance. Sociodemographic characteristics and physical activities were investigated using a standard questionnaire delivered by face-to-face interview. BMDs were 0.48 +/- 0.01 and 0.37 +/- 0.11 g/cm2 in men and women, respectively. In men, age, weight, body mass index (BMI), LBM, FM, physical activity, smoking, alcohol, and education were significantly correlated with BMD. In women, age, weight, BMI, LBM, FM, education, years since menopause, number of deliveries, and number of children breast-fed were significantly correlated with BMD. By multiple regression, LBM, education, smoking, and alcohol in men, and age, LBM, FM, smoking, and number of delivery in women were independent determinants of BMD. LBM was an important contributor for BMD in men, but both LBM and FM were equally important contributors in female to BMD. This stems from the fact that body composition changes with age differ in men and women. Thus, the augmentation of muscle mass in men and the maintenance of an optimal weight in women act to prevent osteoporosis. PMID- 15336619 TI - Effect of discontinuation of alendronate treatment and exercise on bone mass and physical fitness: 15-month follow-up of a randomized, controlled trial. AB - The purpose of this study was to evaluate the remaining effects of 12-month intervention of alendronate and exercise on selected risk factors of fragility fractures in 15-month follow-up after withdrawal of intervention among early postmenopausal women. The trial consisted four experimental groups: (1) 5 mg of alendronate daily + exercise (Al+Ex+), (2) 5 mg alendronate daily (Al+Ex-), (3) placebo + exercise (Al-Ex+), and (4) placebo (Al-Ex-). At the follow-up measurements, bone mass and physical fitness of 102 women (mean age 53.5 +/- 2.5 years) out of initial 150 subjects could be evaluated. Alendronate increased bone mass significantly [mean; 95% confidence interval (CI)] during the intervention at the lumbar spine (3.9%; 2.2% to 5.7%) and femoral neck (2.1%; 0.9% to 3.4%). After withdrawal of alendronate, bone loss resumed to the rate comparable to that evident in the placebo group. Despite the declining bone mass, the between-group mean difference (3.2%; 1.0% to 5.4%) remained at the lumbar spine. However, the benefits at the femoral neck had disappeared 15 months after the withdrawal of alendronate. The 12-month exercise training resulted in significant increases in muscle power, dynamic balance, and aerobic capacity with no benefits on bone mass. Fifteen months later, these performance variables had declined among both the exercisers and nonexercisers. Although the between-group differences were no longer statistically significant, muscle power, dynamic balance, and aerobic capacity of those who exercised still remained above the pretraining levels. In conclusion, 12-month treatment with alendronate prevented postmenopausal bone loss, and residual effect was seen 15 months after withdrawal of the drug at the lumbar spine. Similarly, exercise improved muscle power, agility, and aerobic capacity during the intervention, but the improvement was lost after the cessation of the exercise program. Based on these results, it was evident that to maintain the benefits of alendronate or exercise, therapy should be continued. PMID- 15336620 TI - Stress fracture injury in young military men and women. AB - Approximately 5% of all military recruits incur stress fracture injuries during intense physical training, predominately in the lower extremity. We compared young men and women with stress fracture injury (subjects) to a matched group of uninjured volunteers (controls) during a summer training program at the United States Naval Academy to identify possible risk factors for stress fracture injury. The subject group was composed of 13 female and 18 male plebes with training-induced stress fracture injury verified by plain radiographs and/or nuclear bone scan. The control group was composed of 13 female and 18 male plebes who remained without injury during plebe summer training but who were matched with the 31 injured plebes for the Initial Strength Test (1-mi run time, means: women, 7.9 min; men, 6.4 min) and body mass index (means: women, 23.4; men, 23.8). We found that the subjects lost significant body weight (mean, 2.63 +/- 0.54 kg) between Day 1 and the date of their diagnosis of a stress fracture (mean, Day 35) and that they continued to lose weight until the date of their DEXA scan (mean, Day 49). Among female plebes, there was no evidence of the female athlete triad (eating disorders, menstrual dysfunction, or low bone density). Thigh girth was significantly smaller in female subjects than in female controls and trended to be lower in male subjects than in male controls. Total body bone mineral content was significantly lower in the male subjects than in male controls. Bone mineral density of the distal tibia and femoral neck were not significantly different between the groups. DEXA-derived structural geometric properties were not different between subjects and controls. Because, on average, tibias were significantly longer in male subjects than in male controls, the mean bone strength index in male subjects was significantly lower than that of male controls. We conclude that significant, acute weight loss combined with regular daily physical training among young military recruits may be a significant contributing risk factor for stress fracture injuries in young military men and women. PMID- 15336621 TI - "Recombomice": the past, present, and future of recombination-detection in mice. AB - Homology directed repair (HDR) provides an efficient strategy for repairing and tolerating many types of DNA lesions, such as strand breaks, base damage, and crosslinks. Recombinational repair and lesion avoidance pathways that involve homology searching are integral to normal DNA replication. Indeed, it is estimated that at least ten HDR events take place each time a mammalian cell divides. HDR is associated with the transfer and exchange of DNA sequences. Usually, homologous sequences are aligned perfectly and flanking sequences are not exchanged. However, those sequence misalignments and exchanges that do occur can lead to rearrangements that contribute to cancer (e.g. deletions, inversions, translocations or loss of heterozygosity (LOH)). In order to reveal genetic and environmental factors that modulate HDR in mammals, several approaches have been used to detect recombination events in vivo. Here, we briefly review three methods for detecting homologous recombination in mice, namely: sister chromatid exchange (SCE), LOH, and recombination at tandem repeats. We conclude with a more detailed description of the recently developed "Fluorescent Yellow Direct Repeat" (FYDR) mouse model, which exploits enhanced yellow fluorescent protein (EYFP) for detecting mitotic homologous recombination in vivo. Applications of the FYDR mice are described, as well as the broader potential for using fluorescent proteins to detect recombination in various tissues/cell types in vivo. PMID- 15336622 TI - Iron chelators increase the resistance of Ataxia telangeictasia cells to oxidative stress. AB - Ataxia telangeictasia (A-T) is an autosomal recessive disorder characterized by immune dysfunction, genomic instability, chronic oxidative damage, and increased cancer incidence. Previously, desferal was found to increase the resistance of A T, but not normal cells to exogenous oxidative stress in the colony forming efficiency assay, suggesting that iron metabolism is dysregulated in A-T. Since desferal both chelates iron and modulates gene expression, we tested the effects of apoferritin and the iron chelating flavonoid quercetin on A-T cell colony forming ability. We demonstrate that apoferritin and quercetin increase the ability of A-T cells to form colonies. We also show that labile iron levels are significantly elevated in Atm-deficient mouse sera compared to syngeniec wild type mice. Our findings support a role for labile iron acting as a Fenton catalyst in A-T, contributing to the chronic oxidative stress seen in this disease. Our findings further suggest that iron chelators might promote the survival of A-T cells and hence, individuals with A-T. PMID- 15336623 TI - Contribution of a conserved phenylalanine residue to the activity of Escherichia coli uracil DNA glycosylase. AB - Uracil DNA glycosylase (UDG) excises uracil from DNA to initiate repair of this lesion. This important DNA repair enzyme is conserved in viruses, bacteria, and eukaryotes. One residue that is conserved among all the members of the UDG family is a phenylalanine that stacks with uracil when it is flipped out of the DNA helix into the enzyme active site. To determine what contribution this conserved Phe residue makes to the activity of UDG, Phe-77 in the Escherichia coli enzyme was mutated to three different amino acid residues, alanine (UDG-F77A), asparagine (UDG-F77N), and tyrosine (UDG-F77Y). The effects of these mutations were measured on the steady-state and pre-steady-state kinetics of uracil excision in addition to enzyme.DNA binding kinetics. The overall excision activity of each of the mutants was reduced relative to the wild-type enzyme; however, each mutation gave rise to a different kinetic phenotype with different effects on substrate binding and catalysis. The excision activity of UDG-F77N was the most severely compromised, but this enzyme still bound to uracil-containing DNA at about the same rate as wild-type UDG. In contrast, the decrease in the excision activity of UDG-F77A is likely to reflect a greater reduction in uracil DNA binding than in the catalytic step. Overall, the effects of the mutations on catalysis are best correlated with the polarity of the substituted residue such that an increase in polarity decreases the efficiency of uracil excision. PMID- 15336624 TI - Relative levels of the two mammalian Rad23 homologs determine composition and stability of the xeroderma pigmentosum group C protein complex. AB - Mammalian cells express two Rad23 homologs, HR23A and HR23B, which have been implicated in regulation of proteolysis via the ubiquitin/proteasome pathway. Recently, the proteins have been shown to stabilize xeroderma pigmentosum group C (XPC) protein that is involved in DNA damage recognition for nucleotide excision repair (NER). Because the vast majority of XPC forms a complex with HR23B rather than HR23A, we investigated possible differences between the two Rad23 homologs in terms of their effects on the XPC protein. In wild-type mouse embryonic fibroblasts (MEFs), endogenous XPC was found to be relatively stable, while its steady-state level and stability appeared significantly reduced by targeted disruption of the mHR23B gene, but not by that of mHR23A. Loss of both mHR23 genes caused a strong further reduction of the XPC protein level. Quantification of the two mHR23 proteins revealed that in normal cells mHR23B is actually approximately 10 times more abundant than mHR23A. In addition, overexpression of mHR23A in the mHR23A/B double knock out cells restored not only the steady-state level and stability of the XPC protein, but also cellular NER activity to near wild-type levels. These results indicate that the two Rad23 homologs are largely functionally equivalent in NER, and that the difference in expression levels explains for a major part the difference in complex formation with as well as stabilization effects on XPC. PMID- 15336625 TI - Nonhomologous end joining of complementary and noncomplementary DNA termini in mouse testicular extracts. AB - Mammalian somatic cells are known to repair DNA double-strand breaks (DSBs) by nonhomologous end joining (NHEJ) and homologous recombination (HR); however, how male germ cells repair DSBs is not yet characterized. We have previously reported the highly efficient and mostly precise DSB joining ability of mouse testicular germ cell extracts for cohesive and blunt ends, with only a minor fraction undergoing terminal deletion [Mutat. Res. 433 (1999) 1]; however, the precise mechanism of joining was not established. In the present study, we therefore tested the ability of testicular extracts to join noncomplementary ends; we have also sequenced the junctions of both complementary and noncomplementary termini and established the joining mechanisms. While a major proportion of complementary and blunt ends were joined by simple ligation, the small fraction having noncleavable junctions predominantly utilized short stretches of direct repeat homology with limited end processing. For noncomplementary ends, the major mechanism was "blunt-end ligation" subsequent to "fill-in" or "blunting", with no insertions or large deletions; the microhomology-dependent joining with end deletion was less frequent. This is the first functional study of the NHEJ mechanism in mammalian male germ cell extracts. Our results demonstrate that testicular germ cell extracts promote predominantly accurate NHEJ for cohesive ends and very efficient blunt-end ligation, perhaps to preserve the genomic sequence with minimum possible alteration. Further, we demonstrate the ability of the extracts to catalyze in vitro plasmid homologous recombination, which suggests the existence of both NHEJ and HR pathways in germ cells. PMID- 15336626 TI - Chloroethylnitrosourea-derived ethano cytosine and adenine adducts are substrates for Escherichia coli glycosylases excising analogous etheno adducts. AB - Exocyclic ethano DNA adducts are saturated etheno ring derivatives formed mainly by therapeutic chloroethylnitrosoureas (CNUs), which are also mutagenic and carcinogenic. In this work, we report that two of the ethano adducts, 3,N4 ethanocytosine (EC) and 1,N6-ethanoadenine (EA), are novel substrates for the Escherichia coli mismatch-specific uracil-DNA glycosylase (Mug) and 3 methyladenine DNA glycosylase II (AlkA), respectively. It has been shown previously that Mug excises 3,N4-ethenocytosine (epsilonC) and AlkA releases 1,N6 ethenoadenine (epsilonA). Using synthetic oligonucleotides containing a single ethano or etheno adduct, we found that both glycosylases had a approximately 20 fold lower excision activity toward EC or EA than that toward their structurally analogous epsilonC or epsilonA adduct. Both enzymes were capable of excising the ethano base paired with any of the four natural bases, but with varying efficiencies. The Mug activity toward EC could be stimulated by E. coli endonuclease IV and, more efficiently, by exonuclease III. Molecular dynamics (MD) simulations showed similar structural features of the etheno and ethano derivatives when present in DNA duplexes. However, also as shown by MD, the stacking interaction between the EC base and Phe 30 in the Mug active site is reduced as compared to the epsilonC base, which could account for the lower EC activity observed in this study. PMID- 15336627 TI - Attempted base excision repair of ionizing radiation damage in human lymphoblastoid cells produces lethal and mutagenic double strand breaks. AB - A significant proportion of cellular DNA damages induced by ionizing radiation are produced in clusters, also called multiply damaged sites. It has been demonstrated by in vitro studies and in bacteria that clustered damage sites can be converted to lethal double strand breaks by oxidative DNA glycosylases during attempted base excision repair. To determine whether DNA glycosylases could produce double strand breaks at radiation-induced clustered damages in human cells, stably transformed human lymphoblastoid TK6 cells that inducibly overexpress the oxidative DNA glycosylases/AP lyases, hNTH1 and hOGG1, were assessed for their radiation responses, including survival, mutation induction and the enzymatic production of double strand breaks post-irradiation. We found that additional double strand breaks were generated during post-irradiation incubation in uninduced TK6 control cells. Moreover, overproduction of either DNA glycosylase resulted in significantly increased double strand break formation, which correlated with an elevated sensitivity to the cytotoxic and mutagenic effects of ionizing radiation. These data show that attempted repair of radiation damage, presumably at clustered damage sites, by the oxidative DNA glycosylases can lead to the formation of potentially lethal and mutagenic double strand breaks in human cells. PMID- 15336628 TI - Spontaneous homologous recombination is decreased in Rad51C-deficient hamster cells. AB - The Chinese hamster cell mutant, CL-V4B that is mutated in the Rad51 paralog gene, Rad51C (RAD51L2), has been described to exhibit increased sensitivity to DNA cross-linking agents, genomic instability, and an impaired Rad51 foci formation in response to DNA damage. To directly examine an effect of the Rad51C protein on homologous recombination (HR) in mammalian cells, we compared the frequencies and rates of spontaneous HR in CL-V4B cells and in parental wildtype V79B cells, using a recombination reporter plasmid in host cell reactivation assays. Our results demonstrate that HR is reduced but not abolished in the CL V4B mutant. We thus, provide direct evidence for a role of mammalian Rad51C in HR processes. The reduced HR events described here help to explain the deficient phenotypes observed in Rad51C mutants and support an accessory role of Rad51C in Rad51-mediated recombination. PMID- 15336629 TI - The rate of extrachromosomal homologous recombination within a novel reporter plasmid is elevated in cells lacking functional ATM protein. AB - Homologous recombination between identical stretches of DNA depends on the coordinated action of many tightly regulated proteins. Cellular defects in homologous recombination are strongly associated with increased genomic instability and tumorigenesis. In cells of the cancer-prone syndrome ataxia telangiectasia (A-T), increased intrachromosomal recombination has been demonstrated, while extrachromosomal recombination has been discussed controversially. We constructed a novel, episomally replicating pGrec recombination vector containing two mutated alleles of the enhanced green fluorescent protein (eGFP) gene. Homologous recombination can reconstitute functional wildtype eGFP, thus allowing detection of recombination events based on cellular eGFP fluorescence. Using an isogenic cell pair of A-T fibroblasts and derivatives complemented by an ATM expression vector, we were able to demonstrate in A-T cells high extrachromosomal recombination rates, which are suppressed upon ectopic ATM expression. We thus found that ATM deficiency increases spontaneous recombination not only in intrachromosomal but also in extrachromosomal substrates, suggesting that lack of ATM increases homologous recombination independent of the chromatin structure. PMID- 15336630 TI - The hyper unequal sister chromatid recombination in an sgs1 mutant of budding yeast requires MSH2. AB - Budding yeast SGS1 and the human Bloom's syndrome (BS) gene, BLM, are homologues of the Escherichia coli recQ. Cells derived from BS patients are characterized by a dramatic increase in sister chromatid exchange (SCE). We previously reported that budding yeast cells deficient in SGS1 showed an increase in the frequency of recombination between unequal sister chromatids recombination (USCR). In this study, we examined the factors influencing the elevated SCR frequency in sgs1 disruptants. The increase in SCR frequency in sgs1 mutants was greatly reduced by disrupting the RAD52 or MSH2 gene, which is involved in mismatch repair. However, a plasmid carrying MSH2, having a missense mutation defective in mismatch repair complemented the reduced USCR in msh2 sgs1 mutants, suggesting that the function of Msh2 in mismatch repair is dispensable for USCR. PMID- 15336631 TI - Identification and characterization of the rlp1+, the novel Rad51 paralog in the fission yeast Schizosaccharomyces pombe. AB - A new DNA repair gene from fission yeast Schizosaccharomyces pombe rlp1+ (RecA like protein) has been identified. Rlp1 shows homology to RecA-like proteins, and is the third S. pombe Rad51 paralog besides Rhp55 and Rhp57. The new gene encodes a 363 aa protein with predicted Mr of 41,700 and has NTP-binding motif. The rlp1Delta mutant is sensitive to methyl methanesulfonate (MMS), ionizing radiation (IR), and camptothecin (CPT), although to a lesser extent than the deletion mutants of rhp55+ and rhp51+ genes. In contrast to other recombinational repair mutants, the rlp1Delta mutant does not exhibit sensitivity to UV light and mitomycin C (MMC). Mitotic recombination is moderately reduced in rlp1 mutant. Epistatic analysis of MMS and IR-sensitivity of rlp1Delta mutant indicates that rlp1+ acts in the recombinational pathway of double-strand break (DSB) repair together with rhp51+, rhp55+, and rad22+ genes. Yeast two-hybrid analysis suggests that Rlp1 may interact with Rhp57 protein. We propose that Rlp1 have an accessory role in repair of a subset of DNA damage induced by MMS and IR, and is required for the full extent of DNA recombination and cell survival under condition of a replication fork collapse. PMID- 15336632 TI - A DNA repair gene of Caenorhabditis elegans: a homolog of human XPF. AB - The xeroderma pigmentosum complementation group F (XPF) protein is a structure specific endonuclease in a complex with ERCC1 and is essential for nucleotide excision repair (NER). We report a single cDNA of Caenorhabditis elegans (C. elegans) encoding highly similar protein to human XPF and other XPF members. We propose to name the corresponding C. elegans gene xpf. Messenger RNA for C. elegans xpf is 5'-tagged with a SL2 splice leader, suggesting an operon-like expression for xpf. Using RNAi, we showed that loss of C. elegans xpf function caused hypersensitivity to ultra-violet (UV) irradiation, as observed in enhanced germ cell apoptosis and increased embryonic lethality. This study suggests that C. elegans xpf is conserved in evolution and plays a role in the repair of UV damaged DNA in C. elegans. PMID- 15336633 TI - Are environmental electromagnetic fields genotoxic? AB - Long-term exposure to extremely-low-frequency electromagnetic fields (ELF EMFs) greater than 0.4 microT has been linked, by epidemiological studies, to a small elevated risk of childhood leukaemia. Laboratory-based experiments have been claimed to show that ELF EMFs induce a variety of biological responses, although these claims are controversial. Recent experiments by Ivancsits et al. [Mutat. Res. 519 (2002) 1; Int. Arch. Occup. Environ. Health 76 (2003) 431; Mech. Age. Dev. 124 (2003) 847; H.W. Rudiger, S. Ivancsits, E. Diem, O. Jahn, Genotoxic effects of ELF-EMF on human cells in vitro, Bioelectromagnetics Society 25th Annual Meeting, Maui, USA, 2003] suggest that ELF EMFs are genotoxic, on the basis of observations that intermittent exposures induce single-strand breaks (SSB) and double-strand DNA breaks (DSB) in the DNA of cultured human fibroblasts. The implications of these findings are discussed. PMID- 15336634 TI - Acupoint stimulation with diluted bee venom (apipuncture) alleviates thermal hyperalgesia in a rodent neuropathic pain model: involvement of spinal alpha 2 adrenoceptors. AB - Chemical acupuncture with diluted bee venom (DBV), termed apipuncture, has been traditionally used in oriental medicine to treat several inflammatory diseases and chronic pain conditions. In the present study we investigated the potential antihyperalgesic and antiallodynic effects of apipuncture in a rat neuropathic pain model. DBV (0.25 mg/kg, subcutaneous) was injected into the Zusanli acupoint 2 weeks after chronic constrictive injury (CCI) of the sciatic nerve. Between 5 and 45 minutes after DBV injection, we observed a significant reduction in the thermal hyperalgesia induced by CCI, but apipuncture failed to reduce CCI-induced mechanical allodynia. We subsequently examined whether this antihyperalgesic effect of apipuncture was related to the activation of spinal opioid receptors and/or alpha2-adrenoceptors. Intrathecal pretreatment with naloxone (10 microg/rat), an opioid receptor antagonist, did not reverse the antihyperalgesic effect of apipuncture, whereas pretreatment with idazoxan (40 microg/rat), an alpha2-adrenoceptor antagonist, completely blocked the effect of apipuncture. These results indicate that DBV-induced apipuncture significantly reduces the thermal hyperalgesia generated by CCI and also suggest that this antihyperalgesic effect is dependent on the activation of alpha2-adrenoceptors, but not opioid receptors, in the spinal cord. PERSPECTIVE: The antinociceptive effect of apipuncture was evaluated in a rodent neuropathic pain model. The relieving effect of apipuncture on thermal hyperalgesia was found to be mediated by spinal alpha2-adrenoceptors, but not opioid receptors. These data suggest that apipuncture might be an effective alternative therapy for patients with painful peripheral neuropathy, especially for those who are poorly responsive to opioid analgesics. PMID- 15336635 TI - Ethnic differences in pain coping: factor structure of the coping strategies questionnaire and coping strategies questionnaire-revised. AB - Coping has been examined extensively in the pain literature, although coping instruments have been typically validated in clinical populations with little ethnic diversity. This study examined the factor structure of the Coping Strategies Questionnaire (CSQ) and the CSQ-Revised (CSQ-R) in 650 healthy male and female African American (44%) and white (56%) subjects and explored associations of coping to health and pain-related measures. Factor analyses revealed 6 components for each ethnic group, accounting for comparable amounts of variance and resembling previously reported CSQ subscales. Internal consistency for both ethnic groups was acceptable (0.72-0.91). There were significant main effects for ethnicity on 4 of the CSQ-R scales (P < .05). No ethnic differences in pain or health variables emerged, although when split into high-pain versus minimal-pain groups, differences were revealed on catastrophizing. Results indicate that the factor structure of the CSQ-R in healthy adults is similar to clinical populations and is comparable across African American and white subjects. Group differences on CSQ-R scales suggest potentially important ethnic influences on pain coping. These findings support the use of the CSQ-R to assess coping in African Americans and in healthy young adults. Additional clinical research is needed to determine the practical importance of group differences in pain coping. PERSPECTIVE: Coping has been examined extensively in the pain literature, although coping instruments typically have been validated in clinical populations with little ethnic diversity. This study examines the factor structure of the CSQ-Revised in an ethnically diverse population and supports the use of the CSQ-R to assess coping in African Americans and in healthy young adults. PMID- 15336636 TI - Population-based survey of pain in the United States: differences among white, African American, and Hispanic subjects. AB - A cross-sectional telephone survey was conducted in nationally representative probability sample of non-Hispanic white subjects, non-Hispanic African American subjects, and Hispanic subjects of any race to explore relationships between chronic pain and race or ethnicity. Approximately one third in each group reported "frequent or persistent pain" for 3 months or longer during the past year, and approximately one third of the 454 white subjects, 447 African American subjects, and 434 Hispanic subjects in the final sample experienced "disabling pain" (defined as both high severity and high functional interference). White subjects had pain longer but with lesser intensity than the other groups, and pain-related life interference did not vary. Significantly fewer Hispanic subjects (68%) than white subjects (82%) or African American subjects (85%) had visited a physician for pain, and African American subjects (81%) were more likely than white subjects (75%) or Hispanic subjects (63%) to have used prescription medications. Disabling pain was positively associated with female sex (odds ratio [OR], 1.45), income of $25,000 or less (OR, 1.71), less than a high school education (OR, 1.72), and divorce (OR, 1.69) and was negatively associated with younger age (18-34 years; OR, 0.68), income between $25,000 and $74,999 (OR, 0.64) or $75,000 or more (OR, 0.37), being employed (OR, 0.48), suburban residence (OR, 0.64), and having a college (OR, 0.51) or graduate (OR, 0.32) degree. Multivariate logistic regression found that income of $25,000 or less (OR, 2.54), less than a high school education (OR, 1.59), and being unemployed (OR, 1.50) remained significant when other factors were controlled. Neither race nor ethnicity predicted disabling pain, but the minorities had more characteristics identified as predictors. The data suggest that race and ethnicity contribute to clinical diversity, but socioeconomic disadvantage is the more important predictor of disabling pain. PERSPECTIVE: Race and ethnicity influence the presentation and treatment of chronic pain. This study evaluated community-dwelling white, African American, and Hispanic subjects by using a sophisticated telephone survey methodology. Pain was highly prevalent across groups, and there were racial and ethnic differences in pain experience and treatment preferences. Race and ethnicity were not independently associated with severe pain, but both minorities were more likely to possess the socioeconomic and educational characteristics that were associated. PMID- 15336637 TI - Modulation of presynaptic activity by phosphorylation in cultured rat spinal dorsal horn neurons. AB - Phosphorylation, in particular by protein kinase C (PKC), modulates spinal sensory transmission and nociceptive behaviors. Whereas PKC's postsynaptic actions are well established, its presynaptic effects in spinal sensory neurons are mostly inferred from postsynaptic recordings. Here we first show that the amphipathic styryl dye FM 1-43 can be used to image presynaptic activity in cultured spinal dorsal horn cultures and then test whether PKC modulates presynaptic activity in cultured spinal dorsal horn neurons. Pretreatment with the broad-spectrum kinase inhibitor staurosporine (2 micromol/L) inhibited dye release. Bisindolylmaleimide I, a PKC inhibitor, potentiated dye release at low doses (200 nmol/L and 1 micromol/L), while inhibiting it at a higher dose (5 micromol/L). Activating PKC with phorbol dibutyrate (0.5 micromol/L) induced an increase in exocytosis, which is partially blocked by bisindolylmaleimide I. These results indicate that styryl dyes can be used to observe presynaptic regulation of spinal dorsal horn neurons, and that PKC acts presynaptically to modulate spinal sensory transmission. PERSPECTIVE: With dye imaging technique, we demonstrate here that PKC presynaptically regulates sensory transmission in spinal dorsal horn neurons. In combination with conventional whole-cell patch clamp recording technique, the present study provides a new methodology for studying spinal sensory transmission and modulation and facilitates our understanding of pain mechanism. PMID- 15336638 TI - Body pain area and pain-related negative affect predict clinical pain intensity in patients with fibromyalgia. AB - Patients with fibromyalgia (FM) report widespread chronic musculoskeletal pain. Palpation of 9 paired tender points (TPs) is commonly used for the diagnosis of FM according to criteria specified by the American College of Rheumatology. Although TP palpation can be used to assess deep tissue hypersensitivity, it has failed as a reliable indicator of clinical pain intensity in FM. The sum of local areas of pain (SLAP) obtained from a body pain diagram represents a relevant measure of the spatial extent of clinical pain, a feature most likely important for FM pain. Because spatial summation of pain can be an important determinant of clinical pain intensity, we hypothesized that this measure would predict clinical pain intensity in FM patients. Because pain is strongly associated with negative emotions, we evaluated the relationship of pain-related negative affect (PRNA) with clinical pain intensity in FM. The independent contributions of SLAP, PRNA, and TP count to the variance of clinical pain intensity were assessed in 280 FM patients. Clinical pain intensity of 280 FM patients was measured by using a visual analogue scale. FM patients shaded all painful body areas on body pain diagrams. Dolorimetry was used for TP evaluations. PRNA was assessed with the Medical College of Virginia Pain Questionnaire. Hierarchical linear regression was used to test the association of SLAP, TPs, and PRNA with clinical pain intensity. FM patients' mean visual analogue scale rating (0 to 100) of usual clinical pain was 50.1. Mean SLAP, TP count, and PRNA were 11.4, 16.0, and 44.3, respectively. Hierarchical linear regression analysis identified SLAP, TP count, and PRNA as independent predictors of clinical pain that accounted for 45% of the variance in clinical pain intensity ratings in FM patients. Consistent with the literature, TP count predicted only a small part (4%) of this variance. Our statistical model of body pain areas and negative affect predicts a large portion of the variance of pain intensity in FM. This result suggests that the extent of pain areas and negative emotions are uniquely associated with clinical pain intensity in FM. PERSPECTIVE: The number of painful body areas obtained by body pain diagrams is a better predictor of clinical pain intensity than TPs in FM patients. The combination of painful body areas, TP counts, and PRNA predicts 45% of the clinical pain intensity of FM patients. This finding might be useful for clinical evaluations of FM patients. PMID- 15336639 TI - Development of a measure of the burden of pain due to herpes zoster and postherpetic neuralgia for prevention trials: adaptation of the brief pain inventory. AB - In preparation for clinical trials of a vaccine against herpes zoster (HZ), we conducted a prospective, observational study to evaluate (1) the Zoster Brief Pain Inventory (ZBPI), an HZ-specific questionnaire to quantify HZ pain and discomfort, (2) an operational definition of postherpetic neuralgia (PHN), and (3) a severity-duration measure of the burden of illness caused by HZ. HZ patients aged 60 years or older (n = 121) were enrolled within 14 days of rash onset and completed ZBPI, McGill Pain Questionnaire Present Pain Intensity (PPI), quality of life (QoL), and activities of daily living (ADL) questionnaires on a predetermined schedule. Reliability, measured by intraclass correlation coefficients within 14 days of rash onset, ranged between 0.63 and 0.78. ZBPI pain scores were strongly correlated with other pain measures, interference with ADL, and worsening QoL. The operational definition of PHN, a ZBPI pain score of 3 or greater occurring 90 or more days after rash onset, had high agreement with pain worse than mild on the PPI (kappa = 0.72). The ZBPI pain severity-duration measure had high correlations with severity-duration measures of ADL interference, worsening QoL, and other pain scales. These findings support the validity and utility of the ZBPI, the definition of PHN, and the severity duration measure of the burden of HZ illness. PERSPECTIVE: Herpes zoster pain, as measured by the ZBPI severity-duration measure, is associated with impairment in daily living activities and quality of life. The ZBPI measure appears useful for quantifying herpes zoster pain, postherpetic neuralgia, and impairment in daily living activities for clinical trials of herpes zoster prevention. PMID- 15336640 TI - Doping, gene transfer and sport. PMID- 15336641 TI - "Genetic Doping" with erythropoietin cDNA in primate muscle is detectable. PMID- 15336642 TI - Therapeutic antibody gene transfer: an active approach to passive immunity. AB - Advances in gene transfer approaches are enabling the possibility of applying therapeutic antibodies using DNA. In particular gene transfer in combination with electroporation is promising and can result in generating in vivo antibody concentrations in the low therapeutic range. However, several important problems need to be dealt with before antibody gene transfer can become a valuable supplement to the current therapies. As antibody production following gene transfer is difficult to control, the danger of inducing autoimmune conditions or uncontrollable side effects occurs in cases in which autologous antigens are targeted. It is suggested that the most promising area of application therefore appears to be infectious disease in which heterologous antigens are targeted and concerns for long-term antibody exposure are minimal. Finally, genes encoding fully human antibodies will enhance long-term expression and decrease problems linked to immunogenicity. PMID- 15336643 TI - Stem cell gene transfer--efficacy and safety in large animal studies. PMID- 15336644 TI - Maximizing antigen targeting to the proteasome for gene-based vaccines. AB - Wild-type or immunoevasive antigens can drive weak CD8+T-cell responses against both dominant and subdominant epitopes during gene-based vaccination. For many antigens, fusion to ubiquitin (Ub) to target them to the proteasome circumvents this problem. Although this procedure works in most cases, for one subset of antigens, Ub fusion does not improve immune responses. To determine why these failures occur, we have evaluated in detail the 'rules' for proteasome targeting that have been applied in mammalian vaccine studies, but that were actually defined in yeast systems. To do this, we fused a series of engineered Ub genes to green fluorescent protein (GFP) and tested their ability to target GFP to the proteasome for enhanced antigen processing and CD8+ T-cell responses. Here we demonstrate that Ub fusion mediates enhanced CD8+ responses by proteasome targeting rather than by enhancing protein translation. We also show that several of the yeast-defined Ub constructs failed to target the proteasome in mammalian cells and likewise failed to enhance transgene-specific CD8+ T-cell responses in mice. In contrast, when mammalian-optimized constructs were applied to target the influenza virus nucleoprotein, CD8+ responses were enhanced against its refractory subdominant epitope in mice. This work demonstrates that Ub fusion has efficacy to enhance CD8+ responses, especially against subdominant antigen epitopes, provided constructs are optimized for mammalian use. PMID- 15336645 TI - Factors influencing the efficacy, longevity, and safety of electroporation assisted plasmid-based gene transfer into mouse muscles. AB - Intramuscular injection of plasmid is a potential alternative to viral vectors for the transfer of therapeutic genes into skeletal muscle fibers. The low efficiency of plasmid-based gene transfer can be enhanced by electroporation (EP) coupled with the intramuscular application of hyaluronidase. We have investigated several factors that can influence the efficiency of plasmid-based gene transfer. These factors include electrical parameters of EP, optimal use of hyaluronidase, age and strain of the host, and plasmid size. Muscles of very young and mature normal, mdx, and immunodeficient mice were injected with plasmids expressing beta galactosidase, microdystrophin, full-length dystrophin, or full-length utrophin. Transfection efficiency, muscle fiber damage, and duration of transgene expression were analyzed. The best transfection level with the least collateral damage was attained at 175-200 V/cm. Pretreatment with hyaluronidase markedly increased transfection, which was also influenced by the plasmid size and the strain and the age of the mice. Even in immunodeficient mice, there was a significant late decline in transgene expression and plasmid DNA copies, although both still remained relatively high after 1 year. Thus, properly optimized EP assisted plasmid-based gene transfer is a feasible, efficient, and safe method of gene replacement therapy for dystrophin deficiency of muscle but readministration may be necessary. PMID- 15336646 TI - In vivo selection of genetically modified erythroid cells using a jak2-based cell growth switch. AB - Cell-based therapies have potential widespread applications in clinical medicine, and methods for controlling the fate of transplanted cells are needed. We have previously described a means for directing the growth of genetically modified cells in vivo using a derivative of the thrombopoietin receptor, mpl, that is reversibly activated by a drug called a chemical inducer of dimerization (CID). Since Jak2 participates in signaling from a number of different cytokine receptors (including mpl), we tested whether direct activation of the JH1 domain of Jak2 would broaden the repertoire of hematopoietic lineages responsive to the CID. While the engineered Jak2 induced a significant rise in genetically modified red cells, as we have observed previously with mpl, it lacked mpl's ability to expand genetically modified platelets and failed to expand genetically modified granulocytes, B cells, or T cells. These findings identify a signaling molecule other than mpl that can function as a cell growth switch in vivo and demonstrate that signaling molecules used for in vivo selection need not be confined to receptors. The erythroid-restricted growth response suggests that CID-activated Jak2 may be well suited to gene therapy applications in sickle cell anemia or beta-thalassemia. PMID- 15336647 TI - High-level in vivo gene marking after gene-modified autologous hematopoietic stem cell transplantation without marrow conditioning in nonhuman primates. AB - The successful engraftment of genetically modified hematopoietic stem cells (HSCs) without toxic conditioning is a desired goal for HSC gene therapy. To this end, we have examined the combination of intrabone marrow transplantation (iBMT) and in vivo expansion by a selective amplifier gene (SAG) in a nonhuman primate model. The SAG is a chimeric gene consisting of the erythropoietin (EPO) receptor gene (as a molecular switch) and c-Mpl gene (as a signal generator). Cynomolgus CD34+ cells were retrovirally transduced with or without SAG and returned into the femur and humerus following irrigation with saline without prior conditioning. After iBMT without SAG, 2-30% of colony-forming cells were gene marked over 1 year. The marking levels in the peripheral blood, however, remained low (<0.1%). These results indicate that transplanted cells can engraft without conditioning after iBMT, but in vivo expansion is limited. On the other hand, after iBMT with SAG, the peripheral marking levels increased more than 20-fold (up to 8-9%) in response to EPO even at 1 year posttransplant. The increase was EPO-dependent, multilineage, polyclonal, and repeatable. Our results suggest that the combination of iBMT and SAG allows efficient in vivo gene transduction without marrow conditioning. PMID- 15336648 TI - Widespread correction of lysosomal storage following intrahepatic injection of a recombinant adeno-associated virus in the adult MPS VII mouse. AB - Mucopolysaccharidosis type VII is a lysosomal storage disease caused by deficiency of the acid hydrolase beta-glucuronidase. MPS VII mice develop progressive lysosomal accumulation of glycosaminoglycans within multiple organs, including the brain. Using this animal model, we investigated whether gene transfer mediated by a recombinant adeno-associated virus (rAAV) type 2 vector is capable of reversing the progression of storage in adult mice. We engineered an rAAV2 vector to carry the murine beta-glucuronidase cDNA under the transcriptional direction of the human elongation factor-1alpha promoter. Intrahepatic administration of this vector in adult MPS VII mice resulted in stable hepatic beta-glucuronidase expression (473 +/- 254% of that found in wild type mouse liver) for at least 1 year postinjection. There was widespread distribution of vector genomes and beta-glucuronidase within extrahepatic organs. The level of enzyme activity was sufficient to reduce lysosomal storage within the liver, spleen, kidney, heart, lung, and brain. Within selected regions of the brain, neuronal, glial, and perivascular cells had histopathologic evidence of reduced storage. Also, brain alpha-galactosidase and beta-hexosaminidase enzyme levels, secondarily elevated by the storage abnormality, were normalized. These data demonstrate that peripheral administration of an rAAV2 vector in adult MPS VII mice can lead to transgene expression levels sufficient for improvements in both the peripheral and the central manifestations of this disease. PMID- 15336649 TI - Long-term correction of ornithine transcarbamylase deficiency by WPRE-mediated overexpression using a helper-dependent adenovirus. AB - The urea cycle disorders (UCDs) are important models for developing gene replacement therapy for liver diseases. Long-term correction of the most common UCD, ornithine transcarbamylase (OTC) deficiency, has yet to be achieved in clinical or preclinical settings. The single human clinical trial using early generation adenovirus (Ad) failed to show any biochemical correction. In adult OTC-deficient mice, an E1/E2-deleted Ad vector expressing the mouse OTC gene, but not the human, was only transiently therapeutic. By using post-transcriptional overexpression in the context of the less immunogenic helper-dependent adenoviral vector, we achieved metabolic correction of adult OTC-deficient mice for >6 months. Demonstrating this result were normalized orotic aciduria, normal hepatic enzyme activity, and elevated OTC RNA and protein levels in the absence of chronic hepatotoxicity. Overexpressing the human protein may have overcome two potential mechanisms accounting for poor cross-species complementation: a kinetic block at the level of mitochondrial import or a dominant negative effect by the mutant polypeptide. These data represent an important approach for treating human inborn errors of hepatocyte metabolism like the UCDs that require high-level transduction and gene expression for clinical correction. PMID- 15336650 TI - Adenovirus-mediated erythropoietin production by airway epithelia is enhanced by apical localization of the coxsackie-adenovirus receptor in vivo. AB - In well-differentiated human airway epithelia, the coxsackie B and adenovirus types 2 and 5 receptor (CAR) resides on the basolateral membrane. Replacing the transmembrane and cytoplasmic tail of CAR with a glycosyl-phosphatidylinositol anchor (GPI-CAR) allows apical localization of GPI-CAR, where it can bind adenovirus and enhance gene transfer in vitro. To test this hypothesis further and to investigate requirements and barriers we developed an in vivo model that quantitatively assesses gene transfer of erythropoietin (EPO) to mouse airway epithelia. Our data suggest that erythropoietin is secreted basolaterally, allowing possible access to the bloodstream. The data also suggest that basolateral adenovirus-mediated airway epithelia EPO secretion persists for long periods and could be used to study persistence in vivo. Additionally, the increase in hematocrit in response to the increased serum EPO could be used for therapeutic purposes. Finally, we tested the ability of apically localized CAR to enhance the infection of AdEPO in mouse airway epithelia in vivo. The data suggest that apical receptors in airway epithelia may be sufficient to improve adenovirus infection of airway epithelia in vivo. PMID- 15336652 TI - Intravesical Ad-IFNalpha causes marked regression of human bladder cancer growing orthotopically in nude mice and overcomes resistance to IFN-alpha protein. AB - We have produced prolonged, high local concentrations of interferon in vivo by intravesical instillation of adenoviruses encoding interferon-alpha (Ad-IFNalpha) together with the gene transfer-enhancing agent Syn3. We found sustained interferon protein levels for days, both in normal mouse urothelium and in human bladder cancer cells growing as superficial bladder tumors in nude mice using an orthotopic bladder model developed by us. Tumor burden in the bladder was determined utilizing cancer cells containing the green fluorescent protein. Marked tumor regression was observed following two 1-h exposures of Ad IFNalpha/Syn3 and little or no cytotoxicity was detected in normal cells. Similar intravesical instillation of clinically relevant concentrations of IFN protein alone or Ad-IFNalpha without Syn3 was ineffective. Surprisingly, in vitro, Ad IFNalpha also caused caspase-dependent death of bladder cancer cell lines that were resistant to high concentrations of IFN-alpha protein, including the cell line used in vivo. These findings demonstrate that Ad-IFNalpha can overcome resistance to IFN-alpha protein both in vitro and in vivo and support evaluation of intravesical Ad-IFNalpha/Syn3 for the treatment of superficial bladder cancer. PMID- 15336651 TI - Differentiation and transcription factor gene therapy in experimental parkinson's disease: sonic hedgehog and Gli-1, but not Nurr-1, protect nigrostriatal cell bodies from 6-OHDA-induced neurodegeneration. AB - We tested the activity of the dopaminergic neuron differentiation factor sonic hedgehog, its downstream transcription factor target Gli-1, and an orphan nuclear receptor, Nurr-1, necessary for the induction of the dopaminergic phenotype of nigrostriatal neurons, in an in vivo model of nigrostriatal neurodegeneration. Our preliminary experiments demonstrated that all three constructs expressed the proper molecules and that these had the predicted biological activities in vitro. We expressed the N-terminal of sonic hedgehog (ShhN) and the Gli-1 and Nurr-1 entire coding regions from highly purified, and quality controlled, replication defective adenoviral vectors injected into the brains of rats and used the dopaminergic growth factor GDNF as a positive control. The neurotoxin 6 hydroxydopamine was used to lesion the nigrostriatal dopaminergic innervation; RAd-ShhN and RAd-Gli-1 protected dopaminergic neuronal cell bodies in the substantia nigra, but not axonal terminals in the striatum, from 6-OHDA-induced cell death, while RAd-Nurr-1 was ineffective in protecting either cell bodies or axons. RAd-GDNF was able to protect both the dopaminergic cell bodies and the striatal axon terminals. Our results establish for the first time, to the best of our knowledge, that gene transfer of ShhN and one of its target transcription factors can selectively protect dopaminergic nigrostriatal neuronal cell bodies from a specific neurotoxic insult. Selective protection of nigrostriatal dopaminergic cell bodies by the differentiation factor ShhN and the transcription factor Gli-1 was achieved in a neurotoxic model that eliminates more than 70% of the nigral neurons under consideration. Differentiation and transcription factors can thus be used for the treatment of neurodegeneration by gene therapy. PMID- 15336653 TI - Potential utility of BimS as a novel apoptotic therapeutic molecule. AB - We have previously demonstrated a 1000-fold induction of gene expression exclusive to Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC) cells using an adenoviral vector (ad5.oriP). This platform allows us to explore tumor-specific gene therapy with BimS (ad5.oriP.BimS), a potent proapoptotic Bcl 2 family member. Ad5.oriP.BimS (25 infectious units (ifu)/cell) reduced C666-1 viability in a time- and dose-dependent manner to 15% survival. The effect was enhanced with radiation (6 Gy). Three days after infection, the proportion of apoptotic cells increased from 3.5% (control) to 47.5% (25 ifu/cell). Confocal microscopy demonstrated Bim colocalization to the mitochondria within 18 h of ad5.oriP.BimS infection. Ad5.oriP.BimS induced a 2.8-, 2.1-, and 1.85-fold increase in caspase-3, -8, and -9 activities, respectively. When C666-1 cells were infected with ad5.oriP.BimS (20 ifu/cell), no tumors formed in 7/9 mice followed for 100 days. Six intratumoral injections of ad5.oriP.BimS (1.5 x 10(9) ifu/dose) in combination with radiation were sufficient to cause almost complete disappearance of established C666-1 or C15 xenograft tumors. Intravenous injections of ad5.oriP.BimS (10(9) ifu) induced mild perturbation in liver function tests, associated with hepatocyte apoptoses and mitoses. This vector appears to be safe and effectively cytotoxic to EBV-positive NPC cells both in vitro and in vivo, mediated primarily through the induction of apoptosis. PMID- 15336654 TI - Noninvasive imaging of enhanced prostate-specific gene expression using a two step transcriptional amplification-based lentivirus vector. AB - Noninvasive evaluation of gene transfer to specific cells or tissues will allow for long-term, repetitive monitoring of transgene expression. Tissue-specific promoters that restrict the expression of a transgene to tumor cells play a vital role in cancer gene therapy imaging. In this study, we have developed a third generation HIV-1-based lentivirus vector carrying a prostate-specific promoter to monitor the long-term, sustained expression of the firefly luciferase (fl) reporter gene in living mice. The fl gene in the transcriptionally targeted vector is driven by an enhanced prostate-specific antigen promoter in a two-step transcriptional amplification (TSTA) system. The efficiency of the lentivirus (LV TSTA)-mediated gene delivery, cell-type specificity, and persistence of gene expression were evaluated in cell culture and in living mice carrying prostate tumor xenografts. In vivo bioluminescence imaging with a cooled charge-coupled device camera revealed significantly high levels of fl expression in prostate tumors. Injection of LV-TSTA directly into the prostate of male nude mice revealed efficient and long-term fl gene expression in the prostate tissue for up to 3 months. These studies demonstrate the significant potential of TSTA-based lentivirus vectors to confer high levels of tissue-specific gene expression from a weak promoter, while preserving cell-type specificity and the ability to image noninvasively the sustained, long-term expression of reporter genes in living animals. PMID- 15336655 TI - Oncolytic vaccinia virus expressing the human somatostatin receptor SSTR2: molecular imaging after systemic delivery using 111In-pentetreotide. AB - Oncolytic vaccinia viruses (VV) have demonstrated tumor specificity, high levels of transgene expression, and anti-tumor effects. The ability to visualize vector biodistribution noninvasively will be necessary as gene therapy vectors come to clinical trials, and the creation of a VV that can both treat tumors and permit noninvasive imaging after systemic delivery is therefore an exciting concept. To facilitate imaging, a VV expressing the human somatostatin receptor type 2 (SSTR2) was created. Cells infected with the SSTR2-expressing VV or controls were incubated with the somatostatin analog 111In-pentetreotide with or without an excess of nonradiolabeled pentetreotide. The SSTR2-infected cells bound 111In pentetreotide sixfold more efficiently than control virus-infected cells and this binding was specifically blocked by nonradiolabeled pentetreotide. Nude mice bearing subcutaneous murine colon CA xenografts were injected intraperitoneally with the SSTR2-expressing VV or control VV. After 6 days, mice were injected with 111In-pentetreotide and imaged. Mice were sacrificed and organs collected and counted in a gamma counter. The uptake of radioactivity in tumors and normal tissues (percentage injected dose per gram) and tumor-to-normal tissue ratios were determined. Tumors infected with the SSTR2-expressing VV accumulated significantly higher concentrations of radioactivity compared to tumors in animals receiving the control virus. SSTR2-infected tumors were visible on imaging 6 days after VV injection and could be visualized for up to 3 weeks post viral injection using repeat injections of 111In-pentetreotide. This reporter gene imaging strategy could be a very effective method to visualize vector distribution, expression, and persistence over time and enhances the potential of VV as a novel anti-cancer therapeutic. PMID- 15336656 TI - Effects of CFTR, interleukin-10, and Pseudomonas aeruginosa on gene expression profiles in a CF bronchial epithelial cell Line. AB - Mutations in CFTR lead to a complex phenotype that includes increased susceptibility to Pseudomonas infections, a functional deficiency of IL-10, and an exaggerated proinflammatory cytokine response. We examined the effects of CFTR gene correction on the gene expression profile of a CF bronchial epithelial cell line (IB3-1) and determined which CF-related gene expression changes could be reversed by IL-10 expression. We performed microarray experiments to monitor the gene expression profile of three cell lines over a time course of exposure to Pseudomonas. At baseline, we identified 843 genes with statistically different levels of expression in CFTR-corrected (S9) cells compared to the IB3-1 line or the IL-10-expressing line. K-means clustering and functional group analysis revealed a primary up-regulation of ubiquitination enzymes and TNF pathway components and a primary down-regulation of protease inhibitors and protein glycosylation enzymes in CF. Key gene expression changes were confirmed by real time RT-PCR. Massive reprogramming of gene expression occurred 3 h after Pseudomonas exposure. Changes specific to CF included exaggerated activation of cytokines, blunted activation of anti-proteases, and repression of protein glycosylation enzymes. In conclusion, the CFTR genotype changes the expression of multiple genes at baseline and in response to bacterial challenge, and only a subset of these changes is secondary to IL-10 deficiency. PMID- 15336657 TI - Methotrexate and cytarabine inhibit progression of human lymphoma in NOD/SCID mice carrying a mutant dihydrofolate reductase and cytidine deaminase fusion gene. AB - An SFG-based retroviral bicistronic vector containing a double-mutant dihydrofolate reductase-cytidine deaminase fusion cDNA (F/S DHFR-CD) with IRES eGFP confers resistance to both methotrexate (MTX) and cytarabine (ara-C). Two weeks after transplantation with marrow transduced with either a fusion or a control gene (eGFP-IRES-NeoR), human lymphoma (SKI-DLCL-1) cells were injected sc into the flanks of nonobese diabetic/severe combined immune deficiency mice. In mock-transplanted mice, maximal tolerated dose (MTD) of posttransplant MTX/ara-C (15/10 mg/kg/day, x3) was unable to control tumor growth. Transfer of the fusion gene allowed doses of MTX/ara-C (25/15 mg/kg/day, x4) twofold higher than the MTD to be tolerated. The tumor burden defined the efficiency of posttransplant chemotherapy; early treatment, 48 h after tumor inoculation, provided tumor-free survival, while starting treatment after having palpable tumor growth (7 days) delayed tumor growth a median time of 28 days. In addition, the early treated group had higher gene expression in peripheral blood and marrow cells than the late treated group (P < 0.05), suggesting that early treatment allowed for enrichment of transduced marrow progenitors. These results encourage clinical studies using this retroviral fusion gene construct. PMID- 15336658 TI - Transactivator and structurally optimized inducible lentiviral vectors. AB - Lentiviral vectors offer well-recognized advantages as a gene delivery system both for the analysis of gene function and as a vehicle for gene therapy. In the present study optimized HIV-1-based vector systems that display efficient doxycycline (Dox)-dependent transgene expression in vitro and in vivo have been developed through the modification of factors that contribute to basal activity levels. Dissection of HIV-1 vectors harboring a tTA-dependent transgene expression cassette revealed several mechanisms that account for Dox-independent transgene expression, including those mediated by an internal CMV promoter, as well as a potential contribution from fusion proteins generated by translational readthrough. A precipitous reduction in basal activity levels was accomplished by separating the transactivator and the transgene cassettes into a binary vector system and by relocating the inducible promoter to the U3 region of the LTR. In addition, substituting the VP16 portion of tTA with the human p65 transactivating domain improved Dox-dependent transgene expression in a number of cell types. Optimizing HIV-1-based vectors culminated in a "toolbox" of vectors suitable for transgene delivery in vitro and in vivo, as conveyed by our ability to control the Dox-dependent differentiation of embryonic fibroblasts into muscle cells in vitro and transgene expression in rat brains. PMID- 15336659 TI - Rapid assessment of anti-HIV siRNA efficacy using PCR-derived Pol III shRNA cassettes. AB - Identification of sequences within a target mRNA that are susceptible to potent siRNA knockdown often requires testing several independent siRNAs or shRNA expression cassettes. Using RNAi against HIV RNAs is further complicated by the length of the viral genome, the complexity of splicing patterns, and the propensity for genetic heterogeneity; consequently, it is most important to identify a number of siRNA targets that potently block viral replication. We previously described a facile PCR-based strategy for rapid synthesis of si/shRNA expression units and their testing in mammalian cells. Using this approach, which is rapid and inexpensive, it is possible to screen a number of potential RNAi targets in HIV to identify those that are most susceptible to RNAi. We report that shRNA expression cassettes constructed by PCR and cotransfected directly into mammalian cells with HIV proviral DNA express shRNAs that are inhibitory to HIV-1 replication. Our results also demonstrate that there is a wide range of efficacies among shRNAs targeting different sites throughout the HIV genome. By screening several different targets we were able to identify a sequence in a common tat/rev exon that is exquisitely sensitive to RNAi. Furthermore we relate the efficacies of our PCR product expressed shRNAs to the relative stabilities of the siRNA duplexes and the accessibilities of the target sites to antisense base pairing in cell extracts. PMID- 15336660 TI - Differential myocardial gene delivery by recombinant serotype-specific adeno associated viral vectors. AB - Recombinant cross-packaging of adeno-associated virus (AAV) genome of one serotype into other AAV serotypes has the potential to optimize tissue-specific gene transduction and expression in the heart. To evaluate the role of AAV1 to 5 virion shells on AAV2 transgene transduction, we constructed hybrid vectors in which each serotype capsid coding domain was cloned into a common vector backbone containing AAV2 replication genes. Constructs were tested for expression in: (1) adult murine heart in vivo using direct injection of virus, (2) neonatal and adult murine ventricular cardiomyocytes in vitro, and (3) adult human ventricular cardiomyocytes in vitro, using green fluorescent protein (GFP) as the measurable transgene. Serotype 1 virus demonstrated the highest transduction efficiency in adult murine cardiomyocytes both in vitro and in vivo, while serotype 2 virus had the greater transduction efficiency in neonatal cardiomyocytes in vitro. Prolonged in vivo myocardial GFP expression was observed for up to 12 months using serotype 1 and 2 vectors only. In human cardiomyocytes, serotype 1 vector was superior in transduction efficiency, followed by types 2, 5, 4, and 3. These data establish a hierarchy for efficient serotype-specific vector transduction in myocardial tissue. AAV1 serotype packaging results in more efficient transduction of genes in the murine and human adult heart, compared to other AAV serotypes. Our results suggest that adult human cardiac gene therapy may be enhanced by the use of serotype 1-specific AAV vectors. PMID- 15336661 TI - Molecular identification of mycorrhizal fungi in Neuwiedia veratrifolia (Orchidaceae). AB - We here apply a previously described method for identification of single peloton orchid mycorrhiza to a key orchid group and extend the usefulness in the heterobasidiomycetes of an existing fungal database for identification of mycorrhizal fungi. We amplified and sequenced mitochondrial ribosomal large subunit DNA from fungi in roots of Neuwiedia veratrifolia (Orchidaceae), a member of the small subfamily Apostasioideae that is sister to the remainder of Orchidaceae, and used the extended database to identify the mycorrhizal fungi. Sequences from fungi cultured from Neuwiedia roots and from direct peloton amplifications were analyzed cladistically with sequences determined from reference fungal collections and published sequences. The fungi from Neuwiedia are referred to the heterobasidiomycetous orders Tulasnellales and Ceratobasidiales, indicating that apostasioids utilize the same fungi as other photosynthetic orchids. The majority of Neuwiedia mycobionts came together in a clade with Tulasnella species, but some were most closely related to Thanatephorus. In some cases members of these two clades were isolated from the same orchid plant, providing another example of multiple mycobionts occurring in a single plant. PMID- 15336662 TI - Rapid radiation and cryptic speciation in squat lobsters of the genus Munida (Crustacea, Decapoda) and related genera in the South West Pacific: molecular and morphological evidence. AB - Squat lobsters (genus Munida and related genera) are among the most diverse taxa of western Pacific crustaceans, though several features of their biology and phylogenetic relationships are unknown. This paper reports an extensive phylogenetic analysis based on mitochondrial DNA sequences (cytochrome c oxidase subunit I and 16S rRNA) and the morphology of 72 species of 12 genera of western Pacific squat lobsters. Our phylogenetic reconstruction using molecular data supports the recent taxonomic splitting of the genus Munida into several genera. Excluding one species (M. callista), the monophyly of the genus Munida was supported by Bayesian analysis of the molecular data. Three moderately diverse genera (Onconida, Paramunida, and Raymunida) also appeared monophyletic, both according to morphological and molecular data, always with high support. However, other genera (Crosnierita and Agononida) seem to be para- or polyphyletic. Three new cryptic species were identified in the course of this study. It would appear that the evolution of this group was marked by rapid speciation and stasis, or certain constraints, in its morphological evolution. PMID- 15336663 TI - Phylogenetic position of Platanista gangetica: insights from the mitochondrial cytochrome b and nuclear interphotoreceptor retinoid-binding protein gene sequences. AB - The evolutionary relationship of peculiar and poorly known Ganges River dolphin with extinct and extant cetaceans has been in the state of confusion for more than a century. The close resemblance of platanistidae with some of the extinct taxon viz., Dalpiaziniidae and Waipatiidae and their sister group relationship with many of the extant lineages of cetaceans has been reported but none of the alternative hypotheses provide an unambiguous placement for this species. The present study provides insights into the molecular relationships of Platanista with other cetaceans based on comprehensive analyses of the mitochondrial cytochrome b and nuclear interphotoreceptor retinoid-binding protein gene sequences, obtained from 15 specimens of Ganges dolphin from India and Bangladesh. The mean substitution distance analysis of phylogenetically informative characters in the cytochrome b sequences suggested that Platanista gangetica is significantly closer (P<0.001) to Mysticeti than to any other group of toothed whales. However, the conventional methods of phylogenetic reconstruction supported this finding with low to moderate (41-69%) bootstrap values. PMID- 15336664 TI - Phylogenetic analysis of Hordeum (Poaceae) as inferred by nuclear rDNA ITS sequences. AB - Hordeum (Poaceae, Triticeae) occurs with 31 species worldwide in temperate regions, with the exception of Australasia. About 50% of the species are polyploids (4x, 6x) or occur as di- and polyploid forms. To analyze the phylogenetic relationships among diploid and polyploid taxa of the genus the nuclear rDNA internal transcribed spacer region (ITS) was analyzed for 91 accessions, representing all Hordeum species, together with 10 outgroup species. PCR products were either directly sequenced (outgroups) or cloned and eight clones per individual were analyzed. Phylogenetic analysis revealed four major clades that concur with the four genome groups in Hordeum (H, I, Xa, and Xu). Allopolyploids, putative autopolyploids, and species groups within the closely related H-genome clade could be identified. The ITS data indicate times of independent evolution of paralogous rDNA clusters on different chromosomes intermitted by sweeps of homogenization among these clusters and bi-directional homogenization of the clusters in diploids. Penalized likelihood analysis revealed an age of about 12 million years (my) for the genus and indicated the start of a rapid radiation in the H-genome group about 2.5 my ago in South America and Asia. PMID- 15336665 TI - Defining relationships between the known members of the cytochrome P450 3A subfamily, including five putative chimpanzee members. AB - An analysis of the cytochrome P450 3A subfamily (CYP3A) was undertaken in order to define relationships across species among subfamily members. Some members were excluded due to incomplete sequences, while others were held in abeyance because of their almost complete homology. This is the first publication of five chimpanzee CYP3A genes-CYP3A4, CYP3A5, CYP3A7, CYP3A43, and CYP3A67. This project utilized two approaches for characterizing possible relationships-phylogenetic analysis and genomic structure. For the phylogenetic analysis, both nucleotide and amino acid sequences were aligned in silico using the CLUSTAL algorithm, and then visually inspected for accuracy. Three different computer software packages were utilized: MEGA 2.1, TREECON 1.3b, and PHYLIP 3.5. Multiple methods were used: neighbor-joining (NJ), minimum evolution (ME), maximum parsimony (MP), and maximum likelihood (ML). The resulting topologies were compared against each other to define the consensus topology. In addition, the chimpanzee, human, mouse, and rat genome databases were searched for intron/exon information pertaining to the included genes. Both methods suggest the same conclusion, defining orthologs is plausible between similar species (i.e., mouse and rat), but is less useful between species of different orders (i.e., primate and rodent) or classes (i.e., mammal and avian). PMID- 15336666 TI - Ascidian molecular phylogeny inferred from mtDNA data with emphasis on the Aplousobranchiata. AB - We explored the usefulness of mtDNA data in assessing phylogenetic relationships within the Ascidiacea. Although ascidians are a crucial group in studies of deuterostome evolution and the origin of chordates, little molecular work has been done to ascertain the evolutionary relationships within the class, and in the studies performed to date the key group Aplousobranchiata has not been adequately represented. We present a phylogenetic analysis based on mitochondrial cytochrome c oxidase subunit I (COI) sequences of 37 ascidian species, mainly Aplousobranchiata (26 species). Our data retrieve the main groups of ascidians, although Phlebobranchiata appeared paraphyletic in some analyses. Aplousobranch ascidians consistently appeared as a derived group, suggesting that their simple branchial structure is not a pleisiomorphic feature. Relationships between the main groups of ascidians were not conclusively determined, the sister group of Aplousobranchiata was the Stolidobranchiata or the Phlebobranchiata, depending on the analysis. Therefore, our data could not confirm an Enterogona clade (Aplousobranchiata+Phlebobranchiata). All of the tree topologies confirmed previous ideas, based on morphological and biochemical characters, suggesting that Cionidae and Diazonidae are members of the clade Aplousobranchiata, with Cionidae occupying a basal position within them in our analyses. Within the Aplousobranchiata, we found some stable clades that provide new data on the evolutionary relationships within this large group of ascidians, and that may prompt a re-evaluation of some morphological characters. PMID- 15336667 TI - Mitochondrial genome data support the basal position of Acoelomorpha and the polyphyly of the Platyhelminthes. AB - We determined 9.7, 5.2, and 6.8 kb, respectively, of the mitochondrial genomes of the acoel Paratomella rubra, the nemertodermatid Nemertoderma westbladi, and the free-living rhabditophoran platyhelminth Microstomum lineare. The identified gene arrangements are unique among metazoans, including each other, sharing no more than one or two single gene boundaries with a few distantly related taxa. Phylogenetic analysis of the amino acid sequences inferred from the sequenced genes confirms that the acoelomorph flatworms (acoels+nemertodermatids) do not belong to the Platyhelminthes, but are, instead, the most basal extant bilaterian group. Therefore, the Platyhelminthes, as traditionally constituted, is a polyphyletic phylum. PMID- 15336668 TI - Molecular phylogenetics, karyotypic diversity, and partition of the genus Myotis (Chiroptera: Vespertilionidae). AB - Nucleotide sequences of the mitochondrial cytochrome b gene are reported from bats of the genus Myotis including species of the endemic southern African subgenus Cistugo, Myotis (Cistugo) sebrai and Myotis (Cistugo) lesueuri. We also examined Myotis annectans from Southeast Asia, and Myotis macropus from Australia. The two species of Cistugo and Myotis annectans represent the only species of Myotis to differ in chromosome number from the common 2n=44 found in >40 species. Our results show that the two species of Cistugo are more divergent from the other species of Myotis than several other well-recognized genera and we recommend elevating Cistugo to full generic rank. Myotis annectans groups well within Myotis, clustering with other Southeast Asian and Japanese species, and thus represents the only species of Myotis known to have diverged from the common 2n=44 karyotype. Myotis macropus clusters within a clade that includes Southeast Asian species. PMID- 15336669 TI - Phylogenetic relationships of Australian and New Zealand feral pigs assessed by mitochondrial control region sequence and nuclear GPIP genotype. AB - Pigs were introduced into Australia and New Zealand in the 18th and 19th centuries, with some establishing feral populations. With few records of pig introductions into these two countries, molecular phylogenetic analysis was used to assess their origins. Mitochondrial (mt) control region sequence and nuclear glucosephosphate isomerase pseudogene (GPIP) restriction fragments were used, as distinct European and Asian domestic pig and Wild Boar control region clades and GPIP genotypes can be recognised. Feral pig control region sequences clustered with either European or Asian domestic pig sequences and both Asian and European GPIP alleles were segregating. It was not possible to distinguish direct importation of Asian domestic animals into Australia and New Zealand from indirect introgression of Asian domestic sequences via Europe. However, the clustering of three feral control region sequences of pigs from northern Australia with Asian Wild Boar implies unrecorded introduction of Wild Boar or crossbred animals into Australia. However, two of these feral pigs had European GPIP alleles. In combination, analyses of control region and GPIP markers suggest that both European and Asian pigs have contributed in similar frequencies to the origins of Australian feral pigs. PMID- 15336670 TI - Molecular phylogeny and biogeography of Oriental voles: genus Eothenomys (Muridae, Mammalia). AB - Oriental voles of the genus Eothenomys are predominantly distributed along the Southeastern shoulder of the Qinghai-Tibetan Plateau. Based on phylogenetic analyses of the mitochondrial cytochrome b gene (1143 bp) obtained from 23 specimens (eight species) of Oriental voles collected from this area, together with nucleotide sequences from six specimens (two species) of Japanese red-backed voles (Eothenomys andersoni and Eothenomys smithii) and five species of the closely related genus Clethrionomys, we revised the systematic status of Eothenomys. We also tested if vicariance could explain the observed high species diversity in this area by correlating estimated divergence times to species distribution patterns and corresponding paleo-geographic events. Our results suggest that: (1) the eight species of Oriental voles form a monophyletic group with two distinct clades, and that these two clades should be considered as valid subgenera--Eothenomys and Anteliomys; (2) Eothenomys eleusis and Eothenomys miletus are not independent species; (3) Japanese red-backed voles are more closely related to the genus Clethrionomys than to continental Asian Eothenomys taxa; and (4) the genus Clethrionomys, as presently defined, is paraphyletic. In addition, the process of speciation of Oriental voles appears to be related to the Trans-Himalayan formation via three recent uplift events of the Qinghai Tibetan Plateau within the last 3.6 million years, as well as to the effects of the mid-Quaternary ice age. PMID- 15336671 TI - A phylogeny of the extant Phocidae inferred from complete mitochondrial DNA coding regions. AB - Despite extensive interest in the systematics of Pinnipedia, questions remain concerning phylogenetic relationships within the Phocidae or "true" seals. Relationships within the phocids and their placement relative to the remaining pinnipeds and major lineages of arctoid carnivores were examined using a large molecular data set consisting of 12 mitochondrial protein coding genes. Phylogenetic analysis including 15 extant species of the Phocidae, and representatives of the Otariidae, Odobenidae, Ursidae, Mustelidae, Canidae, and Felidae confirmed the monophyletic origins of the Pinnipedia within the Arctoidea. Slightly more support was found for an ursid affinity of the pinnipeds, however, this relationship remains contentious. The Phocidae were placed as the sister group to a common odobenid-otariid clade. Within the family Phocidae, strong support for the traditionally accepted subfamilies Phocinae (northern seals), and Monachinae (southern seals plus monk seals) was found. In contrast to recent suggestions, a monophyletic Monachus was strongly supported and was placed in a deep branching position within the Monachinae. Evidence from sequence divergence under a maximum likelihood model illustrated that the rarely used tribal distinction within the Monachinae are comparable, in terms of evolutionary distance, to accepted tribal distinctions within the Phocinae. In addition, results suggest that Pagophilus should be accepted as a genus within the Phocini. Sequence divergence between Phoca, Pusa, and Halichoerus is minimal, supporting a taxonomic reclassification of the three genera into an emended genus Phoca, without subgeneric distinctions. PMID- 15336672 TI - Phylogenetic relationships among Opisthobranchia (Mollusca: Gastropoda) based on mitochondrial cox 1, trnV, and rrnL genes. AB - We reconstructed the phylogenetic relationships among 37 species representing seven main lineages within Opisthobranchia (Mollusca: Gastropoda) based on a mitochondrial fragment that included partial cox 1, complete trnV, and partial rrnL genes (about 2500 bp). Phylogenetic analyses confirmed tentatively that all studied main opisthobranch lineages conformed monophyletic groups except Nudibranchia. The sacoglossan Ascobulla was placed as the most basal lineage of opisthobranchs. The basommatophoran pulmonate Siphonaria was recovered within Opisthobranchia between Ascobulla and the remaining opisthobranchs. The latter were divided into two different lineages that await formal description: on one side, Cephalaspidea, Tylodinoidea, and Anaspidea (sharing features in the reproductive, digestive, and circulatory systems) were grouped together and, on the other Architectibranchia and Nudipleura (sharing similarities in the circulatory system) were recovered as sister group taxa. Two well-supported clades were recovered within Nudipleura: Pleuroanthobranchia (new taxon) and Cladobranchia. Pleuroanthobranchia (Pleurobranchoidea plus Anthobranchia) was defined by the presence of blood gland, the presence of calcareous spicules in the integument and the presence of a caecum with an opening directly into the stomach. The new molecular phylogeny provided a robust framework for comparative studies, and prompted a revision of the morphological synapomorphies diagnosing the main clades within opisthobranchs. PMID- 15336673 TI - Phylogeny and biogeography of Rhododendron subsection Pontica, a group with a tertiary relict distribution. AB - Rhododendron subgenus Hymenanthes subsection Pontica is exceptional among Tertiary relict groups in having a high proportion of species (4 of 11) native to SW Eurasia. A phylogeny based on cpDNA matK and trnL-F indicated that multiple Pontica lineages colonised each of SW Eurasia, SE North America, and NE Asia, with little or no speciation within regions thereafter. Therefore, multiple (3-4) Pontica lineages survived the Quaternary in SW Eurasia, in contrast to other Tertiary relict genera. Pontica comprises two major clades, one of which is wholly Eurasian, and paraphyletic with respect to at least some of the remaining 200 species of subgenus Hymenanthes, which are all distributed in SE Asia. The other clade has species from W and SE North America, SW Eurasia, and NE Asia. According to synonymous matK substitution data, the two clades diverged 9-6 million years ago (mya), whereas most divergence within them happened 5-3 mya. Although the phylogeny indicates probable trans-Atlantic migration for one of two America-Eurasia disjunctions in Pontica, the timing supports migration via Beringia for both. PMID- 15336674 TI - Phylogeny of Iris based on chloroplast matK gene and trnK intron sequence data. AB - Phylogenetic analyses of 46 species of Iris, representing all subgenera and all sections except Regelia, Brevituba, and Monolepis, utilized matK gene and trnK intron sequence data. Sequence data show that Iris is paraphyletic because Belamcanda chinensis is resolved within the genus. The two largest subgenera, Iris and Limniris, are both resolved as polyphyletic. With the removal of section Hexapogon, subgenus Iris is weakly supported as monophyletic. Relationships within subgenus Limniris are more complex with the subgenus as currently circumscribed representing eight independent origins among the species included in this study. Several potential monophyletic groups are identified including subgenus Scorpiris, series Spuria (subgenus Limniris section Limniris), and a clade of section Limniris species from North America and Asia. PMID- 15336675 TI - Phylogeny of caecilian amphibians (Gymnophiona) based on complete mitochondrial genomes and nuclear RAG1. AB - We determined the complete nucleotide sequence of the mitochondrial (mt) genome of five individual caecilians (Amphibia: Gymnophiona) representing five of the six recognized families: Rhinatrema bivittatum (Rhinatrematidae), Ichthyophis glutinosus (Ichthyophiidae), Uraeotyphlus cf. oxyurus (Uraeotyphlidae), Scolecomorphus vittatus (Scolecomorphidae), and Gegeneophis ramaswamii (Caeciliidae). The organization and size of these newly determined mitogenomes are similar to those previously reported for the caecilian Typhlonectes natans (Typhlonectidae), and for other vertebrates. Nucleotide sequences of the nuclear RAG1 gene were also determined for these six species of caecilians, and the salamander Mertensiella luschani atifi. RAG1 (both at the amino acid and nucleotide level) shows slower rates of evolution than almost all mt protein coding genes (at the amino acid level). The new mt and nuclear sequences were compared with data for other amphibians and subjected to separate and combined phylogenetic analyses (Maximum Parsimony, Minimum Evolution, Maximum Likelihood, and Bayesian Inference). All analyses strongly support the monophyly of the three amphibian Orders. The Batrachia hypothesis (Gymnophiona, (Anura, Caudata) receives moderate or good support depending on the method of analysis. Within Gymnophiona, the optimal tree (Rhinatrema, (Ichthyophis, Uraeotyphlus), (Scolecomorphus, (Gegeneophis Typhlonectes) agrees with the most recent morphological and molecular studies. The sister group relationship between Rhinatrematidae and all other caecilians, that between Ichthyophiidae and Uraeotyphlidae, and the monophyly of the higher caecilians Scolecomorphidae+Caeciliidae+Typhlonectidae, are strongly supported, whereas the relationships among the higher caecilians are less unambiguously resolved. Analysis of RAG1 is affected by a spurious local rooting problem and associated low support that is ameliorated when outgroups are excluded. Comparisons of trees using the non-parametric Templeton, Kishino-Hasegawa, Approximately Unbiased, and Shimodaira-Hasegawa tests suggest that the latter may be too conservative. PMID- 15336676 TI - Phylogenetic relationships of the African bush-shrikes and helmet-shrikes (Passeriformes: Malaconotidae). AB - The African bush-shrikes and helmet-shrikes (Malaconotidae sensu [A Complete Checklist of the Birds of the World, third ed., Helm Editions, London, 2003]) include 10 genera and 52 species of predatory passerine birds for which monophyly, sister-group, and inter-generic relationships are disputed. To resolve their relationships, we analyzed 2313 bp of sequence data obtained from two nuclear introns (myoglobin intron-2, beta-fibrinogen intron-5) and a mitochondrial protein-coding gene (ND2) using parsimony, maximum likelihood, and Bayesian inference. A strongly supported clade that included representatives of the Malaconotidae, Platysteiridae, and Vangidae was found in all analyses. Three main groups emerged within this clade but relationships between these three groups were always poorly supported. The first group included the helmet-shrikes (Prionops), flycatcher-shrikes (Bias and Megabyas), and vangas (Cyanolanius and Pseudobias), currently placed in the families Malaconotidae, Platysteiridae, and Vangidae, respectively. The second group consisted of four Platysteiridae genera (Lanioturdus, Batis, Platysteira, and Dyaphorophyia), with the remaining Malaconotidae genera ('core malaconotids') forming the last group. Two main clades emerged within the 'core malaconotids,' with the position of the genus Nilaus being variable. The first clade included Malaconotus, Dryoscopus, Bocagia, and Tchagra and the second Chlorophoneus, Laniarius, Rhodophoneus, and Telophorus. Monophyly of the genus Chlorophoneus was never recovered, a result that is consistent with morphological data. PMID- 15336677 TI - Should we be worried about long-branch attraction in real data sets? Investigations using metazoan 18S rDNA. AB - Although long-branch attraction (LBA) is frequently cited as the cause of anomalous phylogenetic groupings, few examples of LBA involving real sequence data are known. We have found several cases of probable LBA by analyzing subsamples from an alignment of 18S rDNA sequences for 133 metazoans. In one example, maximum parsimony analysis of sequences from two rotifers, a ctenophore, and a polychaete annelid resulted in strong support for a tree grouping two "long branch taxa" (a rotifer and the ctenophore). Maximum-likelihood analysis of the same sequences yielded strong support for a more biologically reasonable "rotifer monophyly" tree. Attempts to break up long branches for problematic subsamples through increased taxon sampling reduced, but did not eliminate, LBA problems. Exhaustive analyses of all quartets for a subset of 50 sequences were performed in order to compare the performance of maximum likelihood, equal-weights parsimony, and two additional variants of parsimony; these methods do differ substantially in their rates of failure to recover trees consistent with well established, but highly unresolved phylogenies. Power analyses using simulations suggest that some incorrect inferences by maximum parsimony are due to statistical inconsistency and that when estimates of central branch lengths for certain quartets are very low, maximum-likelihood analyses have difficulty recovering accepted phylogenies even with large amounts of data. These examples demonstrate that LBA problems can occur in real data sets, and they provide an opportunity to investigate causes of incorrect inferences. PMID- 15336678 TI - Evaluating alternative hypotheses for the origin of eusociality in corbiculate bees. AB - We use a likelihood-based statistical test to evaluate the extent to which the available molecular data sets can be used to falsify alternative phylogenetic hypotheses describing the inter-relationship among corbiculate bee tribes. Based on the results of this test, we explore three alternative models of behavioural character state evolution and evaluate the support each model has for single origin versus dual-origin hypotheses for 'highly' eusocial behaviour. We show that only one of four data sets could statistically reject any of the 15 possible outgroup-rooted phylogenetic hypotheses. However, a cytochrome b data set rejected all but three alternative topologies. Using this information, a simple model of behavioural character state evolution, in which transitions between solitary/communal, 'primitively' eusocial, and 'highly' eusocial are unconstrained, supports single-origin hypotheses for 'highly' eusocial behaviour, in spite of phylogenetic uncertainty. By contrast, an ordered model, in which 'highly' eusocial is constrained to be an evolutionarily terminal state, supports a dual-origins hypothesis. Our results show that the molecular phylogenetic evidence favouring a dual-origins hypothesis for 'highly' eusocial behaviour is, at present, conditional on information from one gene (cyt b) and on specific, though likely realistic, assumptions regarding the nature of eusocial evolution. PMID- 15336679 TI - Dracula ant phylogeny as inferred by nuclear 28S rDNA sequences and implications for ant systematics (Hymenoptera: Formicidae: Amblyoponinae). AB - Ants are one of the most ecologically and numerically dominant families of organisms in almost every terrestrial habitat throughout the world, though they include only about 1% of all described insect species. The development of eusociality is thought to have been a driving force in the striking diversification and dominance of this group, yet we know little about the evolution of the major lineages of ants and have been unable to clearly determine their primitive characteristics. Ants within the subfamily Amblyoponinae are specialized arthropod predators, possess many anatomically and behaviorally primitive characters and have been proposed as a possible basal lineage within the ants. We investigate the phylogenetic relationships among the members of the subfamily, using nuclear 28S rDNA sequence data. Outgroups for the analysis include members of the poneromorph and leptanillomorph (Apomyrma, Leptanilla) ant subfamilies, as well as three wasp families. Parsimony, maximum likelihood, and Bayesian analyses provide strong support for the monophyly of a clade containing the two genera Apomyrma+Mystrium (100% bpp; 97% ML bs; and 97% MP bs), and moderate support for the monophyly of the Amblyoponinae as long as Apomyrma (Apomyrminae) is included (87% bpp; 57% ML bs; and 76% MP bs). Analyses did not recover evidence of monophyly of the Amblyopone genus, while the monophyly of the other genera in the subfamily is supported. Based on these results we provide a morphological diagnosis of the Amblyoponinae that includes Apomyrma. Among the outgroup taxa, Typhlomyrmex grouped consistently with Ectatomma, supporting the recent placement of Typhlomyrmex in the Ectatomminae. The results of this present study place the included ant subfamilies into roughly two clades with the basal placement of Leptanilla unclear. One clade contains all the Amblyoponinae (including Apomyrma), Ponerinae, and Proceratiinae (Poneroid clade). The other clade contains members from subfamilies Cerapachyinae, Dolichoderinae, Ectatomminae, Formicinae, Myrmeciinae, and Myrmicinae (Formicoid clade). PMID- 15336680 TI - Phylogeography of surface and cave Astyanax (Teleostei) from Central and North America based on cytochrome b sequence data. AB - Astyanax fasciatus has become a model organism for the study of regressive and adaptive evolution in cave animals. To fully understand these processes, it is important to have background information on the systematics and phylogeography of surface and cave populations of this species. Here we investigate the phylogeography of A. fasciatus in North and Central America and also the historical biogeography of this region. Phylogenetic analysis of part of the mtDNA cytochrome b gene from 26 surface and nine cave A. fasciatus populations revealed seven major clades, which, in principle, represent geographical patterns of distribution. However, the four strongly eye and pigment reduced cave populations, Piedras, Sabinos, Tinaja, and Curva, form a separate cluster, which is not sister group to the surface populations from the same locality. Similarly the Belizean populations do not cluster with their geographic neighbors from the Yucatan. The analyses indicate that there have been recurrent invasions of surface Astyanax from the south, that were most likely influenced by major climate changes during the Pleistocene. During this period, ancestors of the strongly eye and pigment reduced cave populations were able to survive underground as thermophilic relics when the surface populations became extinct. The high level of genetic divergence among the different clades shows that differing haplotype lineages must have reinvaded the surface waters from the south and/or back-colonized them from residual habitats and also penetrated into the caves. Nested clade analyses show that recurrent gene flow as well as historic processes like past fragmentation and range expansion have influenced current populations of A. fasciatus in Central and North America. Different haplotype clades of the phylogeny are not compatible with the present taxonomy of Astyanax and, therefore, we propose the application of a single systematic unit, called A. fasciatus. PMID- 15336681 TI - Phylogeny and intraspecific variability of holoparasitic Orobanche (Orobanchaceae) inferred from plastid rbcL sequences. AB - The rbcL sequences of 106 specimens representing 28 species of the four recognized sections of Orobanche were analyzed and compared. Most sequences represent pseudogenes with premature stop codons. This study confirms that the American lineage (sects. Gymnocaulis and Myzorrhiza) contains potentially functional rbcL-copies with intact open reading frames and low rates of non synonymous substitutions. For the first time, this is also shown for a member of the Eurasian lineage, O. coerulescens of sect. Orobanche, while all other investigated species of sects. Orobanche and Trionychon contain pseudogenes with distorted reading frames and significantly higher rates of non-synonymous substitutions. Phylogenetic analyses of the rbcL sequences give equivocal results concerning the monophyly of Orobanche, and the American lineage might be more closely related to Boschniakia and Cistanche than to the other sections of Orobanche. Additionally, species of sect. Trionychon phylogenetically nest in sect. Orobanche. This is in concordance with results from other plastid markers (rps2 and matK), but in disagreement with other molecular (nuclear ITS), morphological, and karyological data. This might indicate that the ancestor of sect. Trionychon has captured the plastid genome, or parts of it, of a member of sect. Orobanche. Apart from the phylogenetically problematic position of sect. Trionychon, the phylogenetic relationships within sect. Orobanche are similar to those inferred from nuclear ITS data and are close to the traditional groupings traditionally recognized based on morphology. The intraspecific variation of rbcL is low and is neither correlated with intraspecific morphological variability nor with host range. Ancestral character reconstruction using parsimony suggests that the ancestor of O. sect. Orobanche had a narrow host range. PMID- 15336682 TI - Phylogenetic relationships of cyphelloid homobasidiomycetes. AB - The homobasidiomycetes includes the mushroom-forming fungi. Members of the homobasidiomycetes produce the largest, most complex fruiting bodies in the fungi, such as gilled mushrooms ("agarics"), boletes, polypores, and puffballs. The homobasidiomycetes also includes species that produce minute, cup- or tube shaped "cyphelloid" fruiting bodies, that rarely exceed 1-2 mm diameter. The goal of this study was to estimate the phylogenetic placements of cyphelloid fungi within the homobasidiomycetes. Sequences from the nuclear large subunit (nuc-lsu) ribosomal DNA (rDNA), 5.8S rDNA, and internal transcribed spacers (ITS) 1 and 2 were obtained for 31 samples of cyphelloid fungi and 16 samples of other homobasidiomycetes, and combined with published sequences. In total, 71 sequences of cyphelloid fungi were included, representing 16 genera. Preliminary phylogenetic analyses of a 1477-sequence data set and BLAST searches using sequences of cyphelloid forms as queries were used to identify taxa that could be close relatives of cyphelloid forms. Subsequent phylogenetic analyses of one data set with 209 samples represented by nuc-lsu rDNA sequences (analyzed with parsimony) and another with 38 samples represented by nuc-lsu and 5.8S rDNA sequences (analyzed with parsimony and maximum likelihood) indicated that cyphelloid forms represent a polyphyletic assemblage of reduced agarics (euagarics clade, Agaricales). Unconstrained tree topologies suggest that there have been about 10-12 origins of cyphelloid forms, but evaluation of constrained topologies with the Shimodaira-Hasegawa test suggests that somewhat more parsimonious scenarios cannot be rejected. Whatever their number, the multiple independent origins of cyphelloid forms represent striking cases of parallel evolutionary reduction of complex fungal morphology. PMID- 15336683 TI - A molecular phylogeny of Oxya (Orthoptera: Acridoidea) in China inferred from partial cytochrome b gene sequences. AB - The grasshoppers of the genus Oxya are well known to damage rice, sugar cane, and other crops, yet their phylogenetic relationships have not been examined with molecular data. In this study, we obtained the 432 bp DNA sequences of the mitochondrial cytochrome b gene from 91 individuals of nine Oxya species and two outgroups (Gesonula punctifrons and Acrida cinerea). Phylogenetic analyses for the molecular data set were then carried out using the maximum parsimony and neighbor-joining methods. The results showed that the nine Oxya species form four well-supported clades, which include (1) O. intricata and O. flavefemura; (2) O. japonica and O. bicingula; (3) O. agavisa; and (4) O. chinensis, O. brachyptera, O. adentata, and O. hainanensis, respectively. In particular, the monophyly of O. hainanensis and O. agavisa is strongly supported, respectively. However, O. flavefemura and O. intricata, O. bicingula, and O. japonica form paraphyletic groups, respectively, and O. chinensis, O. adentata, and O. brachyptera form a polyphyletic group, suggesting that they should be merged as few as three species. PMID- 15336684 TI - Therapeutic efforts in Duchenne muscular dystrophy; the need for a common language between basic scientists and clinicians. AB - Major advances in molecular genetics of Duchenne dystrophy over the past decade have generated a flurry of attempts at potential cell and gene therapy, mainly in the dystrophin-deficient mdx mouse. This has been accompanied by a fanfare of publicity, in both scientific and lay press, producing waves of hope followed by troughs of disappointment and frustration in both patients and their families and in the scientific community. It has also spawned an additional problem in the use of inappropriate terminology to describe clinical or pathological changes in experimental animal studies, which have been equated with the human disease. It seemed timely to address and hopefully redress the problem, and suggest some solutions, aimed at finding a common language for basic and clinical scientists in their therapeutic efforts in relation to Duchenne dystrophy. Core problems include equating the mdx mouse, with its very mild clinical phenotype, and Duchenne dystrophy; use of inappropriate and often emotive terminology to describe pathological changes, such as 'rescue', 'reversal', 'prevention', 'phenotype', instead of clear descriptive language; and use of the term therapy in place of experiment in both laboratory and clinical experiments targeting single muscles. A major missing link in these multidisciplinary efforts is the absence of mouse clinicians, who can define at a clinical level the motor, respiratory and cardiac deficits in the dystrophic animal, and bridge the huge gap between the mouse scientists doing experimental studies in the laboratory and the clinicians and veterinarians caring for humans and dogs with these disorders. PMID- 15336685 TI - Concerns about the design of clinical trials for spinal muscular atrophy. AB - The distinctive clinical course of SMA, characterized by slowing of the rate of degeneration with the passage of time, presents a special challenge to therapeutic clinical trial planning. Much of the actual functional decline may represent either an inevitable consequence of growth or the result of various secondary complications of weakness, making the study of agents intended to improve the course by increasing the level of SMN protein that much more difficult. Studies intended to demonstrate a slowing of the rate of degeneration, modeled upon clinical trials for ALS, are problematic. In contrast, short-term trials designed to demonstrate improved strength have substantial design advantages, but depend upon the demonstration of salutary effects of increased SMN that are plausible but at present only theoretical. This form of study thus has some potential for type II error, falsely rejecting a useful drug. Despite this limitation, logistic and statistical concerns suggest that the best strategy for evaluating any promising new therapy will be to use first a short-term study. PMID- 15336686 TI - Genotype-phenotype correlations in nemaline myopathy caused by mutations in the genes for nebulin and skeletal muscle alpha-actin. AB - We present comparisons of the clinical pictures in a series of 60 patients with nemaline myopathy in whom mutations had been identified in the genes for nebulin or skeletal muscle alpha-actin. In the patients with nebulin mutations, the typical form of nemaline myopathy predominated, while severe, mild or intermediate forms were less frequent. Autosomal recessive inheritance had been verified or appeared likely in all nebulin cases. In the patients with actin mutations, the severe form of nemaline myopathy was the most common, but some had the mild or typical form, and a few showed other associated features such as intranuclear rods or actin accumulation. Most cases were sporadic, but in addition there were instances of both autosomal dominant and autosomal recessive inheritance, while two families showed mosaicism for dominant mutations. Although no specific phenotype was found to be associated with mutations in either gene, clinical and histological features together with pedigree data may be used in guiding mutation detection. Finding the causative mutation(s) determines the mode of inheritance and permits prenatal diagnosis if requested, but will not as such permit prognostication. PMID- 15336687 TI - Follow-up of nemaline myopathy in two patients with novel mutations in the skeletal muscle alpha-actin gene (ACTA1). AB - Nemaline myopathy has been associated with mutations in five different genes, which all encode protein components of the sarcomeric thin filaments. We report follow-up studies in two children with mutations not previously described in skeletal muscle alpha-actin (ACTA1). Case 1 was a male patient who after birth suffered from pronounced muscle weakness and hypotonia. Muscle biopsy showed small fibers with numerous rods. He failed to achieve any motor milestones. At the age of 17 he required 24 h ventilator support. He could not lift his arms against gravity, but he could use his hands to control his electric wheelchair. The muscle biopsy showed marked replacement of muscle tissue by fat and connective tissue. Only few fibers showed nemaline rods. He had a de novo, heterozygous mutation, G268D in ACTA1. Case 2 was a female patient with feeding difficulties and mild hypotonia in the neonatal period. Muscle biopsy showed hypoplastic muscle fibers and numerous rods. At 11 years of age she walked and moved unhindered and could run fairly well. She had a de novo, heterozygous mutation, K373E, in ACTA1. These two patients illustrate the marked variability in the clinical features of nemaline myopathy in spite of similar muscle pathology in early childhood. The severe muscle atrophy with replacement of fat and connective tissue in case 1 demonstrates the progressive nature of nemaline myopathy in some cases. The described two mutations add to the previously reported mutations in ACTA1 associated with nemaline myopathy. PMID- 15336688 TI - Deflazacort in Duchenne muscular dystrophy: a comparison of two different protocols. AB - We compare the long-term benefits and side effects of deflazacort using two treatment protocols from Naples (N) and Toronto (T). Boys with Duchenne muscular dystrophy between the ages of 8 and 15 years and who had four or more years of deflazacort treatment were reviewed. Diagnostic criteria included males with proximal muscle weakness evident before 5 years, increased serum creatine kinase and genetic testing and/or a muscle biopsy consistent with Duchenne muscular dystrophy. Thirty-seven boys were treated with protocol-N using deflazacort at a dose of 0.6 mg/kg per day for the first 20 days of the month and no deflazacort for the remainder of the month. Boys with osteoporosis received daily vitamin D and calcium. Deflazacort treatment started between 4 and 8 years of age. Thirty two were treated with protocol-T using deflazacort at a dose of 0.9 mg/kg per day, plus daily vitamin D and calcium. Treatment started between 6 and 8 years of age. All boys were monitored every 4-6 months. The results were compared with age matched controls in the two groups (19 for protocol-N and 30 for protocol-T). For the boys treated with protocol-N, 97% were ambulatory at 9 years (control, 22%), 35% at 12 years (control, 0%), 25% at 15 years (control, 0%). For the 32 boys treated with protocol-T, 100% were ambulatory at 9 years (control, 48%), 83% at 12 years (control, 0%) and 77% at 15 years (control, 0%). No aids or leg braces were used for ambulation. In boys 13 years and older, a scoliosis of >20 degrees developed in 30% of the boys on protocol-N, 16% on protocol-T and 90% of controls. For protocol-N, no cataracts were observed while in protocol-T, 30% of boys had asymptomatic cataracts that required no treatment. Fractures occurred in 19% (control 16%) of boys on protocol-N and 16% (control, 20%) of boys on protocol-T. This report illustrates: (a) the importance of collaborative studies in developing treatment protocols in Duchenne muscular dystrophy and (b) the long term beneficial effects of deflazacort treatment in both protocols. However, the protocol-T seems to be more effective and frequently is associated with asymptomatic cataracts. PMID- 15336690 TI - Evolution of the mdx mouse cardiomyopathy: physiological and morphological findings. AB - Heart failure is a major cause of death in boys with Duchenne muscular dystrophy. In order to determine if the cardiac function of the mdx mouse is similarly disturbed, we performed murine echocardiograms and left heart catheterization studies, along with morphometric analysis of cardiac fibrosis. Serial echocardiograms in mdx mice revealed the evolution from normal cardiac function in young mice to a dilated cardiomyopathy in adult mice. Very old mdx mice exhibited a widespread but patchy increase in ventricular wall fibrosis. These results show that the mdx cardiac function is more impaired than was previously thought and shares important clinical features with the cardiomyopathy of Duchenne muscular dystrophy. PMID- 15336689 TI - Prednisolone decreases cellular adhesion molecules required for inflammatory cell infiltration in dystrophin-deficient skeletal muscle. AB - The mechanism of prednisolone's efficacy in the dystrophic pathology is unclear. Prednisolone's anti-inflammatory functions may be particularly important considering the significance of inflammatory cells in dystrophinopathy. In other pathologies, prednisolone's anti-inflammatory effects can be mediated by reducing cellular adhesion molecule (CAM) expression. The goal of this study was to examine the effects of prednisolone on inflammation and CAM expression in dystrophic muscle. Dystrophin-deficient, mdx mice were treated with 0.75 mg/kg prednisolone from 2 to 4 weeks of age. Prednisolone reduced macrophages (-59%, 57%), CD4(+) T-cells (-50%, -60%), CD8(+) T-cells (-58%, -48%), and eosinophils ( 36%, -25%) in quadriceps and soleus muscles, respectively. Prednisolone-treated mice also exhibited decreased vascular P-selectin (-82%) and ICAM-1 (-52%) expression and fewer L-selectin (-79%) and ICAM-1 (-57%) expressing mononuclear cells in quadriceps. Prednisolone reduced sarcolemmal damage and degeneration as well. Our data show that prednisolone is an effective anti-inflammatory in dystrophic muscle and may function by modulating CAM expression. PMID- 15336691 TI - In situ hybridization analysis of Dmpk mRNA in adult mouse tissues. AB - Myotonic dystrophy1 (DM1) is an autosomal dominant, multi-system disorder resulting from a CTG repeat expansion located in the 3' untranslated region of DMPK and immediately in the 5' of SIX5. Skeletal muscle, heart and smooth muscle are prominently affected in DM1. Endocrine abnormalities, gonadal atrophy, brain, skin, skeletal, immune and respiratory defects are also features of the disorder. Both DMPK and SIX5 levels are decreased in DM1 patients. Importantly, expression of mutant mRNAs encoding expanded CUG repeats has been shown to alter the activity of CUG repeat binding proteins in DM1. Mouse models have demonstrated that decreased levels of Dmpk, Six5 and the expression of expanded CUG repeats independently contribute to the development of DM1 pathology. However, an important gap in these studies is a lack of clear understanding of the expression pattern of Dmpk. We demonstrate that Dmpk mRNA is expressed in a range of adult mouse tissues that show pathology in DM1 including skeletal muscle, heart, smooth muscle, bone, testis, pituitary, brain, eye, skin, thymus and lung. Thus DMPK loss or CUG repeat expansion could contribute to DM1 pathology to these tissues. Dmpk mRNA is not detected in the ovary, pancreas or kidney. Significantly, Dmpk mRNA is expressed in the intestinal epithelium, cartilage and liver, which have not been reported to show consistent abnormalities in Dmpk(-/-) mice or in transgenic animals expressing CUG repeats. Taken together these data suggest that Dmpk loss or CUG repeat expression per se may not be sufficient to initiate pathology and are consistent with the hypothesis that coexpression of specific CUG repeat binding proteins with the mutant Dmpk mRNA or deregulation of genes such as Six5 that flank the CTG repeat tract may be necessary for DM1 to manifest. PMID- 15336693 TI - Inducing muscle hypertrophy as a therapeutic strategy for muscular dystrophies. 122nd ENMC International Workshop, Naarden, The Netherlands, 28-30 November 2003. PMID- 15336692 TI - Large-scale gene expression analysis of human skeletal myoblast differentiation. AB - To study pathways involved in human skeletal myogenesis, we profiled gene expression in human primary myoblast cells derived from three individuals using both oligonucleotide and cDNA microarrays. Following stringent statistical testing (false-positive rate 0.4%), we identified 146 genes differentially expressed over time. Interestingly, 86 of these genes have not been reported to be involved in myogenesis in mouse cell lines. This demonstrates the additional value of human primary cell cultures in the study of muscle differentiation. Many of the identified genes play a role in muscle regeneration, indicating the close relationship of this process with muscle development. In addition, we found overlap with expression profiling studies in muscle from Duchenne muscular dystrophy patients, confirming ongoing muscle regeneration in Duchenne muscular dystrophy. Further study of these genes can bring new insights into the process of muscle differentiation, and they are candidate genes for neuromuscular disorders with an as yet unidentified cause. PMID- 15336694 TI - Report on the 124th ENMC International Workshop. Treatment of Duchenne muscular dystrophy; defining the gold standards of management in the use of corticosteroids. 2-4 April 2004, Naarden, The Netherlands. PMID- 15336695 TI - 'Dropped head sign' in myasthenia gravis. PMID- 15336696 TI - Functional analyses of an LXXLL motif in nuclear receptor corepressor (N-CoR). AB - Transcriptional repression is a major regulatory mechanism in cell differentiation, organogenesis, and oncogenesis. Two repressors of ligand dependent transcription factors, nuclear receptor corepressor (N-CoR) and the related protein SMRT were identified as a silencing mediator for thyroid hormone receptor beta and as a silencing mediator for retinoic acid and thyroid hormone receptors, respectively. Nuclear receptor coactivators such as steroid receptor coactivator-1 (SRC-1) contain multiple LXXLL motifs, which are essential and sufficient for its ligand-dependent interaction with nuclear receptors. N-CoR also has an LXXLL motif, located between repressor domains 1 and 2, and conserved between mouse and man. In contrast, SMRT lacks this motif. This paper describes functional implications of the LXXLL motif in N-CoR. A 57-amino acid portion of N CoR containing the LDNLL sequence (N-CoR(LDNLL)) fused to GST interacted with retinoic acid receptor alpha (RARalpha) and thyroid hormone receptor beta (TRbeta) in vitro. Similarly, [(35)S-methionine]N-CoR(LDNLL) interacted with a RARalpha fusion protein. N-CoR(LDNLL) also bound to RARalpha in vivo as determined in mammalian one-hybrid system in transfected CV-1 cells and by two hybrid assays in bacteria. The interaction with RARalpha in vitro and in vivo was specific as determined by mutation of the sequence LDNLL to LDNAA. Our data suggest that the LDNLL motif in N-CoR has functional significance because it mediates interaction with nuclear receptors such as RARalpha and TRbeta. PMID- 15336697 TI - Modulation of transcriptional sensitivity of mineralocorticoid and estrogen receptors. AB - Recent reports describe the ability of factors to modulate the position of the dose-response curve of receptor-agonist complexes, and the amount of partial agonist activity of receptor-antagonist complexes, of androgen, glucocorticoid (GRs), and progesterone receptors (PRs). We now ask whether this modulation extends to the two remaining steroid receptors: mineralocorticoid (MRs) and estrogen receptors (ERs). These studies of MR were facilitated by our discovery that the antiglucocorticoid dexamethasone 21-mesylate (Dex-Mes) is a new antimineralocorticoid with significant amounts of partial agonist activity. Elevated levels of MR, the co-activators TIF2 and SRC-1, and the co-repressor SMRT do modulate the dose-response curve and partial agonist activity of MR complexes. Interestingly, the precise responses are indistinguishable from those seen with GRs in the same cells. Thus, the unequal transactivation of common genes by MRs versus GRs probably cannot be explained by differential responses to changing cellular concentrations of homologous receptor, co-activators, or co repressors. We also find that the dose-response curve of ER-estradiol complexes is left-shifted to lower steroid concentrations by higher amounts of exogenous ER. Therefore, the modulation of either the dose-response curve of agonists or the partial agonist activity of antisteroid, and in many cases the modulation of both properties, is a common phenomenon for all of the classical steroid receptors. PMID- 15336698 TI - Biosynthesis and catabolism of prostaglandin F2alpha (PGF2alpha) are controlled by progesterone in the rat uterus during pregnancy. AB - Myometrial quiescence is a key factor in all species to accomplish a successful gestation. PGs play a crucial role in mediating parturition events, and their synthesis and metabolism are regulated by cyclooxygenases (COXs) and NAD(+) dependent 15-hydroxy-PG dehydrogenase (PGDH), respectively. Progesterone (P(4)) is the hormone responsible for maintaining uterine smooth muscle quiescence during pregnancy. In this work, we have studied the effect of P(4) on the activity of COXs and PGDH, the uterine enzymes involved in the biosynthesis and metabolism of prostanoids in the rat. We found that during pregnancy PGF(2alpha) production and also protein levels of COX-1 and COX-2 were decreased. The exogenous administration of P(4) significantly inhibited the uterine production of PGF(2alpha) and also the protein level of COX-2. PGF(2alpha), metabolism was assessed by PGDH activity, which resulted high during pregnancy and increased as a result of P(4) administration. These results indicate that PGs levels were negatively modulated by P(4), which could be exerting its effect by increasing PGs metabolism through stimulation on PGDH activity and an inhibition on COX and that is a major mechanism for maintain uterine quiescence in pregnancy. PMID- 15336699 TI - The effects of betamethasone (BM) on endothelial nitric oxide synthase (eNOS) expression in adult baboon femoral arterial endothelial cells. AB - Glucocorticoids have significant effects on endothelium mediated vascular function throughout life. The baboon model has been used extensively to study cellular responses to glucocorticoids at several stages of the lifespan. Endothelial nitric oxide synthase (eNOS) is a major regulator of endothelium dependent arterial vasodilation. We have previously demonstrated that synthetic glucocorticoids down regulate eNOS in the baboon placenta. We have now conducted studies to determine whether glucocorticoids would alter eNOS expression in adult systemic vascular endothelial cells in this important animal model. We explored this potential mechanism in endothelial cells from femoral arteries of adult baboons at necropsy and cultured to the fourth passage. Endothelial cells were treated with 10-100nM betamethasone for 24h at 37 degrees C. Vascular endothelial growth factor (VEGF) was used as a positive control and medium as negative controls. The role of glucocorticoid receptor mediation in betamethasone-induced eNOS changes was investigated with the glucocorticoid receptor antagonist mifepristone. RNA (real-time quantitative RT-PCR) and protein (ELISA) were extracted and measured for eNOS. Expression and subcellular distribution of glucocorticoid receptor were detected with fluorescence labeled antibody microscopy. eNOS mRNA and protein in baboon endothelial cells were downregulated 25% by betamethasone treatment. This effect was attenuated by pre-incubation with mifepristone (P < 0.01). VEGF upregulated eNOS transcription and translation (P < 0.001), medium did not alter eNOS expression. We observed that mifepristone and VEGF increased glucocorticoid receptor cytoplasmic accumulation by fluorescence microscopy. We conclude that betamethasone can downregulate eNOS in cultured baboon endothelial cells via the glucocorticoid receptor pathway. PMID- 15336700 TI - Cell type-specific bidirectional regulation of the glucocorticoid-induced leucine zipper (GILZ) gene by estrogen. AB - Estrogen has numerous beneficial physiological actions; however, by acting as a mitogen, it plays a significant role in the induction and maintenance of breast cancer. Although the positive effects of estrogen on gene expression are well described, negative gene regulation is not. Using microarray analysis, we identified 27 genes that were up-regulated and 20 that were down-regulated by estrogen in MCF-7 human breast cancer cells. One gene encoding GILZ (glucocorticoid-induced leucine zipper protein), a putative apoptosis-regulating transcription factor, is rapidly down-regulated by estrogen in these cells. Estrogen antagonists block the down-regulation. The region of the GILZ promoter between nucleotides -104 and -69 mediates both basal activity and estrogen dependent down-regulation in MCF-7 cells. This region contains a functional Oct-1 binding site and a cyclic AMP response element binding protein (CREB) binding site. The same DNA region mediates up-regulation by estrogen in HeLa and HEK293 cells, indicating that cell-specific factors are involved in estrogen regulation of this gene. The estrogen receptor (ER) is present in GILZ promoter protein complexes, but it does not bind directly to the promoter itself, as the DNA binding domain of the estrogen receptor is not required for down-regulation. Elimination of the CREB binding site blocks both basal activity and estrogen regulation. Our results suggest that ER action at the CRE may mediate estrogen dependent, cell-specific regulation of this gene. PMID- 15336701 TI - Estrogenic activity of glabridin and glabrene from licorice roots on human osteoblasts and prepubertal rat skeletal tissues. AB - Data from both in vivo and in vitro experiments demonstrated that glabridin and glabrene are similar to estradiol-17beta in their stimulation of the specific activity of creatine kinase, although at higher concentrations, but differ in their extent of action and interaction with other drugs. In pre-menopausal human bone cells, the response to estradiol-17beta and glabridin (at higher concentration) was higher than in post-menopausal cells; whereas, glabrene (at higher concentration) was more effective in post-menopausal cells. At both ages, the response to estradiol-17beta and glabridin was enhanced by pretreatment with the less-calcemic Vitamin D analog CB 1093 (CB) and the demonstrably non-calcemic analog JK 1624 F(2)-2 (JKF). The response to glabrene was reduced by this pretreatment. Both glabridin and glabrene stimulated creatine kinase specific activity in diaphyseal bone and epiphyseal cartilage of prepubertal female rats. Daily feeding (3-14 days) of prepubertal female rats with glabridin, estradiol 17beta or their combination, also stimulated creatine kinase specific activity. Glabridine, similarly to estradiol-17beta, also stimulated creatine kinase specific activity in ovariectomized female rats. Raloxifene, in combination with glabridin or estradiol-17beta, demonstrated the phenomenon of mutual annihilation of stimulation of creatine kinase specific activity in both epiphysis and diaphysis. Glabrene activity was not inhibited by raloxifene. Therefore, glabridin shows greater similarity to estradiol-17beta and thus greater potential, with or without Vitamin D, to modulate bone disorders in post menopausal women. PMID- 15336702 TI - Partial agonist/antagonist properties of androstenedione and 4-androsten 3beta,17beta-diol. AB - Androgens play important endocrine roles in development and physiology. Here, we characterize activities of two "Andro" prohormones, androstenedione (A-dione) and 4-androsten-3beta,17beta-diol (A-diol) in MDA-MB-453 (MDA) and LNCaP cells. A dione and A-diol, like cyproterone acetate, were partial agonists of transfected mouse mammary tumor virus (MMTV) and endogenous prostate-specific antigen (PSA) promoters. Different from bicalutamide but similar to CPA, both are inducers of LNCaP cell proliferation with only mild suppression of 5alpha-dihydrotestosterone (DHT)-enhanced cell growth. Like bicalutamide and cyproterone acetate, A-dione and A-diol significantly antagonized DHT/R1881-induced PSA expression by up to 30% in LNCaP cells. Meanwhile, in MDA cells, EC(50)s for the MMTV promoter were between 10 and 100nM. Co-factor studies showed GRIP1 as most active for endogenous androgen receptor (AR), increasing MMTV transcription by up to five fold, without substantially altering EC(50)s of DHT, A-dione or A-diol. Consistent with their transcriptional activities, A-dione and A-diol bound full length endogenous AR from MDA or LNCaP cells with affinities of 30-70nM, although binding to expressed ligand-binding domain (LBD) was >20-fold weaker. In contrast, DHT, R1881, and bicalutamide bound similarly to LBD or aporeceptor. Together, these data suggest that A-dione and A-diol are ligands for AR with partial agonist/antagonist activities in cell-based transcription assays. Binding affinities for both are most accurately assessed by AR aporeceptor complex. In addition to being testosterone precursors in vivo, either may impart its own transcriptional regulation of AR. PMID- 15336703 TI - Phosphorylation and function of the hamster adrenal steroidogenic acute regulatory protein (StAR). AB - In order to study the effect of phosphorylation on the function of the steroidogenic acute regulatory protein (StAR), 10 putative phosphorylation sites were mutated in the hamster StAR. In pcDNA3.1-StAR transfected COS-1 cells, decreases in basal activity were found for the mutants S55A, S185A and S194A. Substitution of S185 by D or E to mimic phosphorylation resulted in decreased activity for all mutants; we concluded that S185 was not a phosphorylation site and we hypothesized that mutations on S185 created StAR conformational changes resulting in a decrease in its binding affinity for cholesterol. In contrast, the mutation S194D resulted in an increase in StAR activity. We have calculated the relative rate of pregnenolone formation (App. V(max)) in transfected COS-1 cells with wild type (WT) and mutant StAR-pcDNA3.1 under control and (Bu)(2)-cAMP stimulation. The App. V(max) values refer to the rate of cholesterol transported and metabolized by the cytochrome P450scc enzyme present in the inner mitochondrial membrane. The App. V(max) was 1.61 +/- 0.28 for control (Ctr) WT StAR and this value was significantly increased to 4.72 +/- 0.09 for (Bu)(2)-cAMP stimulated preparations. App. V(max) of 5.53 (Ctr) and 4.82 ((Bu)(2)-cAMP) found for S194D StAR preparations were similar to that of the WT StAR stimulated preparations. At equal StAR quantity, an anti-phospho-(S/T) PKA substrate antibody revealed four times more phospho-(S/T) in (Bu)(2)-cAMP than in control preparations. The intensity of phosphorylated bands was decreased for the S55A, S56A and S194A mutants and it was completely abolished for the S55A/S56A/S194A mutant. StAR activity of control and stimulated preparations were diminished by 73 and 72% for the mutant S194A compared to 77 and 83% for the mutant S55A/S56A/S194A. The remaining activity appears to be independent of phosphorylation at PKA sites and could be due to the intrinsic activity of non phosphorylated StAR or to an artefact due to the pharmacological quantity of StAR expressed in COS-1. In conclusion we have shown that (Bu)(2)-cAMP provokes an augmentation of both the quantity and activity of StAR, and that an enhancement in StAR phosphorylation increases its activity. The increased quantity of StAR upon (Bu)(2)-cAMP stimulation could be due to an augmentation of its mRNA or protein synthesis stability, or both; this is yet to be determined. PMID- 15336704 TI - Specific interactions of steroids, arylhydrocarbons and flavonoids with progesterone receptors from the cytosol of the fungus Rhizopus nigricans. AB - Rhizopus nigricans (R. nigricans) transforms fungitoxic progesterone into the less toxic 11alpha-hydroxyprogesterone which is then able to exit the mycelia into the surrounding water. Hydroxylation of progesterone is an inducible process in which cytosolic progesterone receptors could be involved. In the present study, we characterised receptors with respect to ligand specificity and to their involvement in progesterone induction of hydroxylase. EC(50) values of different ligands (steroids, xenobiotic arylhydrocarbons and natural flavonoids) were determined by competition studies using 40nM ((3)H)progesterone. C21 and C19 3 oxo-4-ene steroids were good competitors (EC(50) of progesterone 2.3 +/- 0.1 x 10(-7)M, EC(50) of androsten-3,17-dione 24 +/- 2 x 10(-7)M). The presence of hydroxyl groups in steroids significantly decreased the affinity for receptors. The arylhydrocarbons alpha-naphthoflavone and ketoconazole exhibited EC(50) values of 0.3 +/- 0.01 x 10(-7)M and 27 +/- 5 x 10(-7)M, respectively, whereas beta-naphthoflavone and benzo(a)pyrene were not able to displace labelled progesterone completely. The competition curves obtained by natural flavonoids also did not reach the bottom level of non-labelled progesterone, indicating the interaction at some allosteric binding site(s) of progesterone receptors. All ligands were examined for their involvement in progesterone-hydroxylase induction. Steroid agonists induced the enzyme in a dose-dependent manner in accordance with their affinity for receptors, whereas arylhydrocarbons and natural flavonoids did not induce the enzyme. The agonistic action of steroids, together with the antagonistic action of alpha-naphthoflavone, strongly suggests the involvement of progesterone receptors in progesterone signalling resulting in the induction of progesterone-hydroxylase. PMID- 15336705 TI - Expression and activation of Akt/protein kinase B in sexually immature and mature rat uterus. AB - This study investigated the expression and activation of Akt/PKB in developing and adult rat uterus. Expression of Akt was observed in uteri from adult ovariectomized and 7-35-day-old rats and no changes were observed in response to in vivo estradiol treatment (1-100 microg/100g b.w.). To examine the mechanisms of PKB/Akt activation, phosphorylation at Thr(308) and Ser(473) regulatory sites were studied in uteri. Akt was constitutively phosphorylated on Ser(473) residue in the untreated, control uteri, while phosphorylation of Thr(308) was observed only after estradiol 17beta (E2) treatment. The effects of E2 treatment were age dependent, no response was induced in 11-day-old uteri, while in 28 days and older rats the activation of Akt at both regulatory sites, Ser(473) and Thr(308), increased, the first response was detected 2h after treatment, reaching the highest rate at 6h. The rate of phosphorylation was stronger at Ser(473) residue. The results suggest that the regulation of Akt activation at two regulatory sites in rat uteri are different, phosphorylation of Thr(308) seems to be entirely estrogen dependent, while the phosphorylation of Ser(473) is regulated by other factors as well as estrogen. PMID- 15336706 TI - Computational systems biology. PMID- 15336707 TI - Levamisole (Decaris) treatment during pregnancy. PMID- 15336708 TI - Epidemiological evidence on reproductive effects of persistent organochlorines in humans. AB - Organochlorines are widespread pollutants in humans. Concern about adverse reproductive effects of these compounds arises from accidental exposure of humans and experimental studies. Recently, this issue has been addressed by a number of studies of exposed populations and hospital-based case-referent studies. These studies indicate that high concentrations of persistent organochlorines may adversely affect semen quality and cause testicular cancer in males, induce menstrual cycle abnormalities and spontaneous abortions in females, and cause prolonged waiting time pregnancy, reduced birth weight, skewed sex ratio, and altered age of sexual development. However, most effects have been demonstrated at exposure levels above the present day exposure level in European and North American populations. Due to inherent methodological problems in several of the available studies, additional research is needed to fully elucidate the possible adverse effects of organochlorines on human reproductive health. PMID- 15336709 TI - Plasticizers, infant nutrition and reproductive health. AB - Since the 1930s, plasticizers have been used to impart flexibility to an otherwise rigid polyvinylchloride (PVC). Di-(2-ethylhexyl)-phthalate (DEHP) is the most widely used plasticizer in PVC formulations. However, DEHP leaks out from PVC items with time and use and consequently it is an ubiquitous environmental contaminant. To date, very little is known about the actual extent of DEHP exposure in infants, who are believed to be the most sensitive population, as they may be exposed to several different sources (breast milk, infant formula, baby food, indoor air, and by dermal and oral exposure via indoor dust containing DEHP) since early in life and since differences in DEHP pharmacokinetics and metabolism between children and adults have been well documented. Little information exists on DEHP dietary exposure, believed to represent a major source and orally administered DEHP is converted more rapidly to MEHP than other routes of exposure. Although DEHP has been shown to induce toxicity in experimental animals, a limited but suggestive human exposure data causes a serious concern that an early in life DEHP exposure may adversely affect male reproductive tract development. Here, we report a review on dietary phthalate exposure in babies. PMID- 15336710 TI - Effects of low doses of dietary lead on puberty onset in female mice. AB - Female Swiss mice typically display signs of puberty at about 33-37 days of age. In the present investigation (96 female mice tested in 8 Pb exposure levels, n=12 per exposure level), the time to puberty onset was markedly influenced by exposure to dietary lead. While modest increases in blood lead concentrations from a normal background of 2-3 to 13.2 microg/dl delayed the onset of puberty by 15-20% to about 40-43 days, reducing blood lead from 2-3 to 0.7 microg/dl was associated with an acceleration of puberty to 21 days, an enhancement by over 30%. This dose-response relationship represents novel findings of possible ecological as well as public health significance and indicates that lead is able to induce biologically significant changes at blood lead levels previously thought to be without effect. PMID- 15336711 TI - Involvement of growth factors in induction of persistent proliferation of vaginal epithelium of mice exposed neonatally to diethylstilbestrol. AB - Neonatal treatment of female mice with natural and synthetic estrogens including diethylstilbestrol (DES) results in persistent proliferation and cornification of vaginal epithelium. In order to study the mechanism of persistent proliferation of vaginal epithelium, histological and biochemical changes were examined in the vagina of C57BL female mice exposed neonatally to 3 microg DES for 5 days. In intact control adult mice, ovariectomy induced apoptotic cell death in vaginal epithelial cells detected by in situ 3'-DNA nick end labeling method accompanied by low DNA synthesis detected by incorporation of bromodeoxyuridine. In neonatally DES-exposed adult mice, however, ovariectomy did not induce reduction of DNA synthesis and showed only a slight increase in apoptotic cells of vaginal epithelium. In neonatally DES-exposed mouse vagina, semi-quantitative reverse transcription polymerase chain reaction revealed a continuous higher expression of mRNAs encoding epidermal growth factor (EGF) and transforming growth factor alpha (TGF-alpha). These results indicate that neonatal DES exposure causes the increase in expression of EGF and TGF-alpha mRNA, possibly resulting in the induction of persistent proliferation and cornification of vaginal epithelium in mice. PMID- 15336712 TI - Neonatal exposure to diethylstilbestrol leads to impaired action of androgens in adult male hamsters. AB - Neonatal treatment with diethylstilbestrol (DES) leads to disruption of spermatogenesis in adult animals after apparently normal testicular development during puberty indicating aberrant androgen action in DES-exposed adult hamsters. The present study determined the effects of exogenous androgens in neonatally DES exposed hamsters. Exogenous androgens failed to reverse the disruption of spermatogenesis in DES-exposed animals. Neonatal DES exposure caused a significant decrease in seminal vesicle weight, and abnormal histology. While exogenous androgens caused a significant increase in seminal vesicle weight in control animals, they failed to restore the seminal vesicle weight and normal histology in DES-exposed animals. Northern blot and/or RT-PCR analysis revealed that (1) AR, ERalpha and ERbeta mRNA levels were unchanged in DES-exposed animals, and (2) mRNA levels for the AR-responsive genes calreticulin, SEC-23B, and ornithine decarboxylase were significantly decreased in DES-exposed animals. Our results suggest that neonatal DES exposure impairs the action of androgens on target organs in male hamsters. PMID- 15336713 TI - Evaluation of the maternal and neonatal effects of the oxytocin antagonist, atosiban, in a cross-fostering study in rats. AB - The purpose of this cross-fostering study was to investigate neonatal survival following exposure of pregnant rats to atosiban (1-deamino-2-D-Tyr(OEt)-4-Thr-8 Orn-oxytocin), an oxytocin antagonist. Atosiban was administered subcutaneously from days 15 to 20 of gestation at 300 mg/kg/day, and controls received vehicle alone. Parturition was observed at 30 min intervals throughout the period that births were occurring. There was no effect of treatment on number of pups born or neonatal viability. Within 1 h of birth, litter size was standardised to five males and five females, followed immediately by cross-fostering either between or within groups. Offspring from treated mothers reared by control mothers had normal survival and weight gain. There was poor survival and weight gain in offspring from control mothers reared by treated mothers. There was clear evidence that lactation was impaired in the treated females, leading to the conclusion that poor neonatal survival in offspring reared by treated mothers was attributable to a failure of milk let-down. PMID- 15336714 TI - Initiation of delayed ovotoxicity by in vitro and in vivo exposures of rat ovaries to 4-vinylcyclohexene diepoxide. AB - Repeated daily dosing of rats with the ovotoxic, occupational chemical 4 vinylcyclohexene diepoxide (VCD, 80 mg/kg, i.p.) selectively depletes primordial and primary ovarian follicles. This study was designed to investigate whether follicle loss can be achieved following a single, acute exposure to VCD. Ovaries removed from postnatal-day-4 female Fischer 344 rats were cultured in the absence or presence of VCD for 15 days. Continuous in vitro exposure to VCD (15 days) caused concentration-dependent loss of primordial and small primary follicles. A single exposure to VCD in vitro (30 microM, 24 h) also caused significant losses of primordial and primary follicles 14 days later. Additionally, 28-day-old female rats were given a single injection of VCD (40-320 mg/kg, i.p.). A single dose at 320 mg/kg resulted in substantial loss of all follicle stages beginning 6 days later. Overall, these results demonstrate that an acute exposure to high concentrations/doses of VCD is sufficient to cause subsequent delayed loss of follicles. PMID- 15336715 TI - Estrogenic and toxic effects of polychlorinated biphenyls on cultured ovarian germ cells of embryonic chickens. AB - Polychlorinated biphenyls (PCBs) are man-made ubiquitous pollutants that have detrimental effects on reproduction and endocrine functions in a variety of species. In the present study, estrogenic and toxic effects of PCBs on embryonic chicken ovarian development were evaluated by a germ-somatic cell co-culture system. Ovarian cells were cultured in serum-free medium and challenged with a mixture of PCBs (Aroclor 1254). Cell proliferation was determined by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) release. Results showed that lower PCBs (0.1-1 microg/ml) manifested mainly estrogenic effect to stimulate germ cell proliferation, while higher PCBs (10 microg/ml) imposed severe toxicity on germ and somatic cells. The toxic effect of PCBs could be attenuated by an antioxidant tocopherol. PCBs induced condensed nuclear chromosome in ovarian cells and caused cell exfoliation and breakdown within initial hours of treatment. After 24 h, the estrogenic effect of PCBs began to exhibit and the survived germ cells manifested proliferation. Inhibition of the estrogenic effect of PCBs by tamoxifen led to increased toxicity on germ cells and somatic cells. These results indicate that PCBs exposure may interfere with ovarian germ cell proliferation and cause reproductive disorder via both toxic and estrogenic actions in embryonic chickens. PMID- 15336716 TI - Effects of gestational and lactational exposure to low doses of PCBs 126 and 153 on anterior pituitary and gonadal hormones and on puberty in female goats. AB - The aim of the present study was to investigate if environmental doses of PCB 153 and PCB 126 could produce effects in a controlled animal model. Possible adverse effects on the hypothalamic-pitutitary-gonadal axis were examined by measuring gonadotrophins and gonadal steroid hormone concentrations in goat kids exposed during gestation and lactation. The concentrations of PCB 153 and PCB 126 in adipose tissue in the goat kids 9 months post-partum were 5800 ng/g (fat-weight, range; 2900-12700 ng/g) and 0.49 ng/g (fat-weight, range; 0.28-0.80 ng/g), respectively. The pre- and post-pubertal plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (Prl) and progesterone (P4) were analysed. LH, FSH, Prl, and P4 were also measured during an induced oestrus cycle. The prepubertal LH concentration was significantly lower, the puberty was delayed and the P4 level during the luteal phase of an estrous cycle was higher in the group exposed to PCB 153. No significant effect of PCB 153 exposure was found on Prl and FSH. PCB 126 did not produce any effects at the exposure level tested in this study. In conclusion, perinatal exposure to PCB 153 affected the reproductive function and the puberty maturation in goats. PMID- 15336717 TI - Differential susceptibility to chromatid breaks induced by bleomycin in sub fertile and fertile bovines. AB - The rate of chromatid breaks was studied in cows with a history of sub-fertility by means of a test based on measurement of the average of breaks induced in lymphocytes of peripheral blood cultures. Fourteen female specimens were divided into two groups: fertile and sub-fertile. Peripheral blood lymphocytes were cultured and prepared for cytogenetic analysis. Two types of culture were established for each animal to evaluate the response of peripheral blood lymphocyte cultures to the genotoxic effects of bleomycin. The first culture did not receive bleomycin treatment (spontaneous chromosome aberrations). Our results showed that median breaks per cell (b/c) (+/-semirange) for spontaneous culture of the fertile and sub-fertile animals and bleomycin sensitivity assay for fertile and sub-fertile animals were 0.00+/-0.06, 0.02+/-0.03, 0.08+/-0.05 and 0.22+/-0.09, respectively. There was no significant difference (P>0.05) in the chromosomal breakage in lymphocytes not exposed to bleomycin; however, in comparing the number of chromatid breaks per cell in cultures treated with bleomycin, the sub-fertile group showed a significantly higher (P<0.05) level than the fertile group. These findings have implications both for identifying cattle with less than optimum fertility as well as for providing potential avenues to study the origins of sub-fertility. PMID- 15336718 TI - Cellular anomalies underlying retinoid-induced phocomelia. AB - The question of how alterations in cell behavior produced by retinoic acid (RA) influenced the development of skeletogenic mesenchyme of the limb bud was examined in this study. Our established model was employed, which involves treatment of pregnant mice with a teratogenic dose of RA (100 mg/kg) on 11 days postcoitum (dpc) resulting in a severe truncation of all long bones of the forelimbs in virtually every exposed fetus. It is shown that RA, administered at a stage to induce phocomelia in virtually all exposed embryos, resulted in immediate appearance of enhanced cell death within the mesenchyme in the central core of the limb bud, an area destined for chondrogenesis. The central core mesenchyme, which in the untreated limb buds experiences a sharp decline in cell proliferation heralding the onset of chondrogenesis, demonstrated a reversal of the process; this mesenchyme maintained a higher rate of cell proliferation upon RA exposure. These events resulted in a truncation and disorganization of the chondrogenic anlage, more pronounced in zeugopodal mesenchyme than in the autopod. We conclude that an inhibition of chondrogenesis was secondary to a disruption in cellular behavior caused by RA, a likely consequence of misregulation in the growth factor signaling cascade. PMID- 15336719 TI - Developmental toxicity of mixtures: the water disinfection by-products dichloro-, dibromo- and bromochloro acetic acid in rat embryo culture. AB - The chlorination of drinking water results in production of numerous disinfection by-products (DBPs). One of the important classes of DBPs is the haloacetic acids. We have previously shown that the haloacetic acids (HAs), dichloro (DCA), dibromo (DBA) and bromochloro (BCA) acetic acid are developmentally toxic in mouse whole embryo culture. Human exposure to these contaminants in drinking water would involve simultaneous exposure to all three HAs. This study explores the question of developmental toxicity interactions between these compounds. Gestational day (GD) 9.5 rat embryos were exposed to various concentrations of the three HAs (singly or in combination) for 48 h and then evaluated for dysmorphology. The embryonic effects from exposure to the single compounds and mixtures were evaluated using developmental score (DEVSC) as the parameter of comparison. Concentrations of individual compounds and mixtures were chosen (based on a dose additivity model) which were predicted to produce scores 10 or 20% lower than control levels. Evaluations were performed on all possible combinations of the three HAs. The HAs were dysmorphogenic and resulted in primarily rotation and heart defects and to a lesser extent prosencephalic, visceral arch, and eye defects. The percent anomalies in the rat were comparable to those previously published for the mouse at comparable toxicant concentration. There was a low incidence of neural tube defects in the rat following exposure to the HAs. The rat neural tube appeared less sensitive to the HAs than did the mouse resulting in a higher rate of neural tube dysmorphology in the mouse. Following exposures to BCA and DBA, alone and in combination, there was a significant incidence of delayed embryonic caudal development with apparent normal development anterior to the second visceral arch. The developmental scores for embryos exposed to combinations of the three compounds, when compared to scores for embryos exposed to the single compounds, indicated that the dose-additivity model adequately predicted the observed toxicity and that the developmental toxicity of these water disinfection by-products appears to be additive in whole embryo culture (WEC). PMID- 15336720 TI - Effect of methylmercury on glutamate-cysteine ligase expression in the placenta and yolk sac during mouse development. AB - The placenta and the yolk sac play critical roles in fetal development, including protection from oxidative stress through the presence of detoxifying enzymes. Glutathione (GSH; gamma-glutamylcysteinylglycine), a crucial molecule in the maintenance of cellular redox status, plays a critical role in development, and it is also protective against methylmercury toxicity. Glutamate-cysteine ligase (GCL), the enzyme that catalyzes the rate-limiting step in GSH synthesis, is widely expressed in the mouse embryo and extraembryonic membranes throughout development. The aim of this study was to investigate the effect of low-level subchronic methylmercury exposure on GCL expression in the mouse placenta and yolk sac, after describing the basal developmental expression of the enzyme in these tissues. We found that basal mRNA expression levels increased dramatically in the placenta and the yolk sac at gd 18, whereas protein levels did not increase in parallel with the mRNA. We also found that methylmercury induced GCLc mRNA expression in the placenta at gd 18 in a dose-dependent manner, suggesting an important role for this enzyme in the response of the placenta to toxicants. These changes in expression may be useful as a biomarker of MeHg exposure during development. PMID- 15336721 TI - Embryotoxicity study of ginsenoside Rc and Re in in vitro rat whole embryo culture. AB - BACKGROUND: Pregnant women commonly consume ginseng. However, there is little data concerning the effects of ginseng on early pregnancy. METHODS: Rat embryos were exposed in vitro to different concentrations of Rc and Re from day 9.5 to day 11.5 after conception. Embryos were scored for growth and differentiation at the end of the culture period. RESULTS: Embryos exposed to 50.0 microg/ml Re had significantly lower median morphological score (29.0 versus 48.0), fewer number of somites (15.0 versus 21.0), and smaller yolk sac diameter (3.5 versus 4.1 mm) and crown-rump length (CRL) (2.9 versus 3.4 mm) compare to control embryos. There was no significant difference between embryos exposed to 5.0 microg/ml Re and control embryos. There was also no difference in the biometric and morphologic parameters among control and embryos exposed to 5.0 and 50.0 microg/ml Rc. CONCLUSION: There is a significant variability in embryotoxic effects of different ginsenosides. Further studies to evaluate the synergistic embryotoxic effects of ginsenosides are warranted. PMID- 15336722 TI - The effects of the reported active metabolite of methoxychlor, 2,2-bis(p hydroxyphenyl)-1,1,1-trichloroethane, on testosterone formation by cultured Leydig cells from young adult rats. AB - Methoxychlor (MC) is an insecticide that is currently used on a variety of agricultural crops, especially following the ban of 2,2-bis(p-chlorophenyl)-1,1,1 trichloroethane (DDT) use in the United States. Following in vivo administration, MC is converted to 2,2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane (HPTE), which is proposed to be the active agent. Both MC and HPTE have been demonstrated to exhibit weak estrogenic and antiandrogenic activities, and they are thought to exert their effects through estrogen or androgen receptors, respectively. A recent study reported that HPTE inhibited both basal and hCG-stimulated testosterone formation by immature and adult cultured rat Leydig cells and that this effect was mediated through the estrogen receptor. In the current studies, we examined the effects of HPTE on basal and hCG-stimulated testosterone formation by cultured Leydig cells from young adult rats. In addition, we evaluated whether the effects of HPTE on rat Leydig cell testosterone biosynthesis were mediated through the estrogen receptor as an estrogen agonist or the androgen receptor as an antiandrogen. The current studies demonstrated that HPTE inhibited both basal and hCG-stimulated testosterone formation in a dose-dependent manner with significant declines in testosterone being observed at approximately 100 nM. The effects of HPTE were localized to the cholesterol side chain cleavage step; however, these effects were not mediated through the classic estrogen receptor or by its acting as an antiandrogen, the currently recognized modes of action of MC and HPTE. PMID- 15336723 TI - Hydrosalpinx fluid induced embryotoxicity and lipid peroxidation. PMID- 15336724 TI - The clinical significance of axillary lymph node micrometastases in breast cancer. AB - OBJECTIVE: To evaluate the clinical significancer of axillary lymph-node micrometastases, in the era of sentinel lymph node (SLN) biopsy. DATA SOURCES: Searches of MEDLINE (1966-2003) and an extensive manual review of journals were performed using the key search terms breast cancer, axillary lymph-node micrometastases, micrometastatic disease, and SLN biopsy. STUDY SELECTION: All articles identified from the data sources were evaluated and all information deemed relevant was included for this review. CONCLUSIONS: Axillary lymph-node micrometastases can be detected by serial sectioning, immunohistochemistry, or reverse transcriptase-polymerase chain reaction (RT-PCR). The presence of axillary SLN micrometastases is generally associated with a worse prognosis and is an indication for axillary lymph node dissection (ALND) and adjuvant therapy. The clinical significance of micrometastases identified by RT-PCR remains unknown and further research with longer follow-up is needed to ascertain the clinical implications of a positive result. PMID- 15336725 TI - The surgical and oncological safety of immediate breast reconstruction. AB - AIM: The aim of our study was to (1) examine the incidence of surgical complications, (2) determine the incidence of loco-regional recurrences and (3) examine the safety of saving the nipple-areola-complex after immediate breast reconstructions in breast cancer. METHODS: Sixty-six immediate breast reconstructions were performed. Wide local excision (WLE), skin sparing mastectomy and subcutaneous mastectomy (SCM) were performed to 12, 20 and 34 patients, respectively. In all patients with WLE the reconstruction was performed with the latissimus dorsi (LD) miniflap. In other patients reconstructions were done with a free TRAM-flap (n=26) LD-flap (n=27) or with a prosthesis only (n=1). RESULTS: Major flap necrosis developed in four patients. Local recurrence rate was 8.3% in the group where nipple-areola-complex was removed and 7.1% in the group where nipple-areola-complex was saved. Metastases were found in 12.5 and 0%, respectively. CONCLUSION: SCM compared to skin sparing mastectomy may lead to an enhanced risk of immediate surgical complications, but does not threat the oncological safety. Saving the nipple-areola-complex in immediate breast reconstructions is possible in early breast cancer, if the tumour is not in the central part of the breast. PMID- 15336726 TI - Residual disease after excision of non-palpable breast tumours: analysis of tumour characteristics. AB - INTRODUCTION: A tumour-positive resection margin is a well-known prognostic factor for local recurrence. The aim of this study was to evaluate tumour characteristics that might be predictive for the presence of residual disease after excisional surgery. PATIENTS AND METHODS: Data of 295 patients, subjected to a wire-guided excisional breast biopsy were studied. Type and size of the primary tumour, the presence of DCIS and an extensive in situ component (EIC), multifocality of the tumour and nodal status were recorded. RESULTS: Residual disease was found in 51% of the patients undergoing a re-operation. 80% of the patients with positive margins were treated by mastectomy. Nodal status and the presence of an extensive in situ component were the only two variables that were statistically significant. CONCLUSION: In case of tumour positive margins axillary involvement and an extensive in situ component in the primary tumour were predictive for residual disease. No subgroups could be defined in whom additional surgery could be omitted. More 'aggressive' surgical therapy is justified in patients belonging to these risk groups. PMID- 15336727 TI - The oncological implications of immediate breast reconstruction. AB - AIMS: The timing of breast reconstruction following mastectomy has been an area of contention. The purpose of this study was to report the oncologic safety of mastectomy and immediate breast reconstruction, primarily with a tissue expander. METHODS: We offered 54 patients (58 reconstructions) an immediate breast reconstruction after a modified radical mastectomy was done for invasive breast cancer. RESULTS: Fifty-eight mastectomies were done for an invasive cancer. Thirteen out of 54 women suffered relapse of their cancer and nine patients died of their disease during the follow-up period. Only in one patient a local recurrence was detected at an early stage and treated without removal of the prosthesis. Disease free survival for all patients was 93% at 5 years. CONCLUSION: Immediate breast reconstruction after mastectomy is an oncologically safe approach and represents a clear improvement in the quality of life for patients with breast cancer. Thus this procedure can safely be offered to appropriately selected women. PMID- 15336728 TI - The potential for improved outcome in patients with hepatic metastases from colon cancer: a population-based study. AB - AIMS: Reports from specialized centres suggest that 20-25% of patients with hepatic metastases from colorectal cancer have resectable disease, with 5-year survival rates of 30-40%, and that an additional 13-38% may become resectable after chemotherapy. The purpose of this study was to assess the potential for improvement in outcome for patients with hepatic metastases from colon cancer in an unselected population. PATIENTS AND METHODS: All patients diagnosed with colon cancer in the Stockholm/Gotland region between 1 January 1996 and 31 December 1999 were identified and followed until 31 December 2002. Treatment and outcome in patients with hepatic metastases was analyzed and CT-scans and MR images of the liver were reviewed to re-evaluate resectability. RESULTS: In 2280 patients with colon cancer, hepatic metastases were diagnosed in 537 patients. Only 21 of these patients underwent a hepatic resection. Retrospective evaluation of liver images indicated that 10% of the patients had potentially resectable hepatic disease. CONCLUSION: The rate of potentially resectable liver metastases from colon cancer in a population is lower than suggested from hospital-based series. With structured management programs including follow-up routines and multidisciplinary treatment protocols the proportion of patients amenable for liver resection may be increased. In this study preoperative chemotherapy might have increased the resectability rate to at the most 17%. To significantly improve prognosis for patients with hepatic metastases from colon cancer more effective treatment modalities are needed. PMID- 15336729 TI - Analysis of prognostic factors after surgery for stage III and IV gallbladder cancer. AB - AIM: Survival and prognostic factors were analysed in patients who had undergone surgical resection with curative intention with the aim of identifying groups of patient with stage III and IV gallbladder cancer on the TNM classification who might benefit from surgery. METHODS: Thirty-seven patients with advanced gallbladder cancer were studied, the cumulative survival rate for each group was calculated for each pTNM factor. RESULTS: The 5-year survival rates in the stage III patients were 83.3%, while those for the stage IVA patients were 46.2%, and those for the stage IVB patients 16.7%. CONCLUSIONS: In patients with invasion of adjacent organs, including the liver and gastrointestinal tract, and rated as pT3 or pT4, extended surgery excising the invaded tissue may be justified. In patients with pN2 lymph-node metastasis, even without adjacent organ invasion, radical surgery may not achieve a good outcome. PMID- 15336730 TI - Analysis of KIT (CD117) expression in gallbladder carcinomas by tissue microarray. AB - AIMS: KIT (CD117) is a transmembrane tyrosinase-kinase receptor which has been related to cell proliferation, differentiation, adhesion and control of apoptosis. If present, KIT may provide a suitable target for tumour therapy. In this study, we report the presence of KIT in primary and metastatic gallbladder carcinomas. METHODS: Formalin-fixed and paraffin-embedded specimens of 57 primary gallbladder carcinomas and 18 corresponding metastases were stained using a tissue microarray technique and two different antibodies. RESULTS: Only three tumours stained for KIT. With a polyclonal antibody only one well differentiated papillary adenocarcinoma was immunoreactive. With a monoclonal antibody two additional poorly differentiated tubular adenocarcinoma showed weak and focal immunostaining. CONCLUSIONS: KIT immunoreactivity is infrequent in gallbladder carcinoma. Thus, routine screening of tumour tissues for KIT by immunohistochemistry appears to be cost-ineffective and cannot be recommended. Moreover, the lack of substantial KIT immunoreactivity in both primary and metastatic gallbladder carcinoma tissues does not provide a rationale to investigate imatinib mesylate therapy in clinical trials including patients with advanced disease. PMID- 15336731 TI - Sentinel-lymph-node biopsy (SLNB) for melanoma is not complication-free. AB - BACKGROUND AND AIM: Sentinel lymph node biopsy (SLNB) is reportedly associated with insignificant morbidity. The study aims at documenting SLNB-related complications and identifying possible risk factors. METHOD: Data of all melanoma patients who underwent SLNB in our medical center (1994-2002) were analysed. Procedure-related complications were recorded. RESULTS: Three hundred and nine lymphatic basins of 250 patients were explored for SLNB. Overall complication rate was 20%. Sensory morbidity was significantly associated with axillary SLNB (p=0.04) and was more prevalent in younger patients. The use of blue dye alone or combined with a hand-held gamma probe had no statistically significant impact on the identification rate. There were six false-negatives (2.3%), for an overall false-negative rate of 18%. A positive sentinel node was significantly associated with shortened overall survival (p=0.04). CONCLUSION: Wound complications are more frequent than usually reported. Sensory morbidity occurs mostly in the axilla. Neck SLNB is associated with the highest rate of identification failure. Patient age, basin location, and number of excised nodes may serve as prognostic factors of morbidity. PMID- 15336732 TI - Surgical treatment of locally recurrent rectal cancer. AB - BACKGROUND: The aim of our study was to analyse data of patients treated by salvage surgery for locally recurrent rectal cancer, with emphasis on the question whether salvage surgery is still worthwhile when adjuvant radiotherapy is no longer a treatment option. METHODS: Forty patients (19 males/21 females) treated by surgery with curative intent for locally recurrent rectal carcinoma were analysed. Local recurrence was defined as cancer recurrence within the lower pelvis. Salvage surgery included abdominoperineal resection, abdominosacral resection, exenteration (posterior or total) and local resection. Clinical and pathological factors were analysed to identify prognostic factors for survival. RESULTS: The median overall survival was 25 months (95% CI: 13-37 months) and 5 year survival was 28% (95% CI: 12-45%). The absence of symptoms at the time of recurrence, central localisation and the absence of microscopic involvement of surgical margins, but not additional radiotherapy, were found to be significant independent prognostic factors for better survival after salvage surgery. CONCLUSION: Salvage surgery, alone or in combination with radiotherapy, can achieve radical resection of locally recurrent rectal cancer and can result in long-term survival. PMID- 15336733 TI - Synchronous extraperitoneal and intraperitoneal dissemination of appendix cancer. AB - BACKGROUND: The aim of this study was to report the role of combined treatments, including cytoreductive surgery and perioperative regional chemotherapy, in patients with synchronous systemic and intraperitoneal dissemination of appendix cancer. METHODS: Patients with synchronous systemic and intraperitoneal dissemination of appendix cancer were treated with cytoreductive surgery and perioperative regional chemotherapy. Variables statistically analyzed with survival as an end point included demographic characteristics, prior surgical score (PSS), tumour marker levels, peritoneal cancer index (PCI), and completeness of cytoreduction (CC). RESULTS: Mean follow up was 42.6 months, median survival time (MST) for 15 patients was 28 months and 5-year survival rate was 29.4%. Female patients had a longer MST than male patients (p=0.0199). Survival was better in patients with PSS 0 and 1 (p=0.0277). Patients with elevated CEA and CA 19-9 levels had a shorter MST (p=0.0083 and p=0.0193, respectively). PCI and CC comparisons did not show significant differences. Morbidity (n=2) and mortality (n=2) rates were 13.3% respectively. CONCLUSION: Acceptable morbidity and mortality and a 29.4% 5-year survival rate allows cytoreductive surgery and regional chemotherapy to be considered as a treatment option for selected patients with synchronous systemic and intraperitoneal dissemination of appendix cancer. PMID- 15336734 TI - Surgery alone in the curative treatment of localised oesophageal carcinoma. AB - AIM: To document the results of surgery alone in patients with localised oesophageal carcinoma. METHODS: Between January 1982 and 2002, 179 consecutive patients who underwent curative oesophagectomy for stage 0, I or II oesophageal carcinoma, without neo-adjuvant treatment, were analysed retrospectively. RESULTS: Postoperative mortality and morbidity rates were 2.8 and 30.7%, respectively. The overall actuarial survival rate at 5 years was 59%. No patients with more than four invaded lymph nodes survived 5 years. A lymph node ratio more than 0.2, more than four invaded lymph nodes and an advanced pTNM stage were predictors of poor prognosis. CONCLUSION: Long-term survival after surgery alone for localised oesophageal carcinoma can be achieved in some 2/3rds of patients. These results should be considered before designing neo-adjuvant therapeutic regimen or embarking into exclusive treatment alternate to oesophagectomy. PMID- 15336735 TI - Primary malignant tumours of bone and soft tissue in the elderly. AB - AIMS: This study reports outcome, functional results and quality of life of 45 elderly patients with age over 70 after surgery for primary malignant bone and soft tissue tumours. METHODS: There were 24 primary malignant bone tumours and 21 soft tissue sarcomas. The most frequent diagnoses were: chondrosarcoma, malignant fibrous histiocytoma and liposarcoma. Local tumour resection with and without osteosynthesis, endoprostheses, and amputations had been performed for surgery. The patients were prospectively followed in a tumour register. RESULTS: Complication and revision rate, functional outcome using the Musculoskeletal Tumor Society score, 5-year survival rate, median survival time and quality of life according to the Life Satisfaction Index A and the global health and quality of-life scale of the QLQ-C30 revealed results that are only slightly inferior to those reported in younger tumour patients. CONCLUSIONS: The results of this study generally justify even extensive tumour surgery in the elderly patient over 70 although outcomes are not quite as good as those reported for younger adults. However, especially in the old patient indications like general condition and comorbidity should be given due consideration before any decision is made on whether surgery should be performed and if so what surgical technique should be applied. PMID- 15336736 TI - The AMORE protocol as salvage treatment for non-orbital head and neck rhabdomyosarcoma in children. AB - AIM: To investigate the feasibility and outcome of the AMORE protocol as salvage treatment in paediatric head and neck rhabdomyosarcoma (HNRMS). METHODS: The AMORE protocol is a local treatment regimen, consisting of Ablative surgery, Moulage technique brachytherapy and surgical Reconstruction, scheduled in 1 week. Patients with recurrent or residual non-orbital HNRMS were eligible for AMORE salvage treatment. RESULTS: The procedure was feasible in nine out of 11 eligible patients. Five patients were treated for recurrent or residual parameningeal RMS after prior chemoradiation. Local complete remission was achieved in all five patients and maintained in four. Three patients are without evidence of RMS with a follow-up duration of 4-10 years. Two patients developed a distant relapse, together with a local recurrence in one. Both patients died of their disease. Four patients were included for recurrent non-parameningeal HNRMS. Long-term local control at the site of recurrence was obtained in all four patients (follow up 5-10 years). CONCLUSIONS: The AMORE protocol is a feasible salvage strategy for non-orbital HNRMS even after external beam radiotherapy. The local salvage rate in this series is promising. PMID- 15336737 TI - Complications following operative treatment of primary malignant bone tumours in the pelvis. AB - AIM: To report the complication rates of limb-salvage reconstruction in the pelvis. Detailed analyses about the type, treatment and outcome of post-operative complications, various reconstruction options are presented. METHODS: Factors that might influence the occurrence of complications were evaluated of 50 consecutive surgically treated patients. RESULTS: The mean follow-up was 57 months. Limb-salvage procedures were used in 42/50 patients, amputations in 8/50 patients. After limb-salvage procedures complications occurred in 32/42 patients, after hemipelvectomy in 6/8 patients. The 1 and 5-year overall survival rate was 92 and 68%, respectively. CONCLUSION: There is a high complication rate in reconstructive techniques using hemipelvic autografts and/or allografts. These procedures are appropriate only in well selected patients. The complication rates following endoprosthetic reconstruction are comparably low. PMID- 15336738 TI - Prevention of radiation enteritis by intrapelvic breast prosthesis. AB - Pelvic malignancies frequently require post-operative radiation therapy that may induce small bowel damage at an incidence of 5-25%. Various surgical techniques have been reported to prevent acute and chronic radiation enteritis. This article describes the technical aspects of pelvic exclusion by an intrapelvic silicone breast prosthesis. PMID- 15336739 TI - Re: The continued utility of needle localised biopsy for non-palpable breast lesion, Hemmer et al. PMID- 15336740 TI - The value of needle localised biopsy for diagnosis and treatment of non-palpable screen detected breast cancer. PMID- 15336741 TI - The two-point discrimination test--time for a re-appraisal? AB - The two-point discrimination (2PD) test is the most frequently used test for the assessment of the sensory outcome after nerve repair. Here we focus on factors which explain the enormous and implausible variability in reported 2PD levels after nerve repair. We conclude that the 2PD testing technique is not at all standardized and that its use as the sole test for tactile gnosis recovery should be seriously questioned. Reports of 2PD results should always be accompanied by a detailed description of how the test was performed, especially with reference to the pressure applied and the testing protocol. PMID- 15336742 TI - Dupuytren's disease risk factors. AB - Dupuytren's is a common problem, but little is known about its aetiology. We have undertaken a large case-control study to assess and quantify the relative contributions of diabetes and epilepsy as risk factors for Dupuytren's in the community. Cases were patients with a diagnosis of Dupuytren's disease and, for each, two controls were individually matched by age, sex, and general practice. Our dataset included 821 cases and 1,642 controls. Five hundred and eighty-eight (72%) of the cases were men. The mean age at diagnosis was 62 (range 24-97) years. Diabetes was a significant risk factor for Dupuytren's disease (OR=1.75) and there was an increased risk for medicinally treated diabetes (metformin- OR=3.56; sulphonylureas--OR=1.75) and particularly insulin controlled (OR=4.38) rather than diet-controlled diabetes. Epilepsy (OR=1.12) and anti-epileptic medications were not associated with Dupuytren's disease. Ascertainment bias in previous studies may explain the reported association with epilepsy. PMID- 15336743 TI - An objective method to evaluate the risk of recurrence and extension of Dupuytren's disease. AB - This study was undertaken to assess the influence of the factors related to Dupuytren's diathesis on the rates of recurrence and extension of Dupuytren's disease after surgery. The records of 65 patients who underwent surgery for Dupuytren's disease were retrospectively studied and the presence of factors related to diathesis were recorded. The sensitivity and specificity of each factor for predicting recurrence and extension were calculated. Odds ratios and 95% confidence intervals were also calculated and a discriminant analysis was performed to explore correlations between recurrence and extension and the significant variables. Our results confirmed the prognostic value of diathesis. The results have been used to develop a new scoring system for evaluating the risk of recurrence and extension. PMID- 15336744 TI - Hand muscle pathology after long-term vibration exposure. AB - The morphology of the abductor pollicis brevis muscle was studied in 20 patients suffering from hand-arm vibration syndrome. The main morphological changes observed were centrally located myonuclei and fibre type grouping (found in all 20 muscle biopsies), angulated muscle fibres (found in 19 biopsies), ring fibres and regenerating fibres (found in 18 biopsies) and fibrosis (found in 17 biopsies). The observed abnormalities are believed to reflect damage to both the muscle fibres and the motor nerve. The changes were related to different vibration exposure parameters. The number of fibres demonstrating centrally located nuclei correlated significantly with the cumulative vibration exposure, while the number of angulated fibres correlated significantly with the total vibration exposure time. This indicates that the vibrating tools may cause direct damage to muscle fibres as well as nerves. PMID- 15336745 TI - Clinical experience with end-to-side nerve transfer. AB - A retrospective study was undertaken to determine the effectiveness of end-to side nerve transfer. Twenty patients with peripheral nerve lesions of varying aetiology underwent 23 end-to-side nerve transfers over a 15-month period. The mean patient age was 30 years and 18 were male. The mean delay in presentation was 2.4 months. All underwent end-to-side nerve repairs and were reviewed in outpatients at regular intervals. Ten patients were lost to follow-up before 12 months and were therefore excluded from the study. The remaining ten patients, who had undergone 13 end-to-side procedures, had a mean follow-up period of 16 months. None demonstrated objective evidence of motor recovery at the end of the study period. Four patients had modest recovery of deep protective sensation, and two patients suffered a subtle degree of "donor" nerve morbidity. We have abandoned this technique in our centre in preference for standard nerve grafting techniques. PMID- 15336746 TI - Gadolinium enhanced MR assessment of proximal fragment vascularity in nonunions after scaphoid fracture: does it predict the outcome of reconstructive surgery? AB - This study investigated whether the outcome of bone graft and internal fixation surgery for nonunion of scaphoid fractures could be predicted by gadolinium enhanced MR assessments of proximal fragment vascularity. Sixteen established scaphoid fracture nonunions underwent gadolinium-enhanced MR scanning before surgical treatment with bone grafting and internal fixation. No relationship was found between MR enhancement and the outcome of surgery. Union was achieved in eight of the 12 nonunions with more than 50% enhancement, and three of the four with less than 50% enhancement, of the proximal pole. Furthermore, union was achieved in both of the nonunions which had less than 25% enhancement of the proximal pole. We conclude that enhanced MR assessments of the vascularity of the proximal fragment of a scaphoid fracture nonunion do not accurately predict the outcome of reconstructive surgery. PMID- 15336747 TI - Arthrodesis of ring finger and little finger metacarpal bases for little finger carpometacarpal joint arthritis. AB - Five patients with localized little finger carpometacarpal arthritis were treated by excision of the little finger metacarpal base and arthrodesis of the little and ring metacarpals. A dorsal periosteal/capsular flap was used as an interposition graft. All patients achieved significant pain relief, good cosmesis and satisfactory grip strength. All returned to activities of daily living. This procedure, the Dubert procedure, is indicated for localized pathology of the hamate-little finger metacarpal joint. It has theoretical advantages over arthrodesis and resection or interposition arthroplasty as it preserves little finger length, rotation and alignment and maintains some mobility of the transverse carpal arch and the little finger ray. PMID- 15336748 TI - Pre-osteotomy plate application technique for ulnar shortening. AB - The self-compressing mode of the AO/ASIF LC-DC plate can be harnessed to close and compress modest osteotomy gaps that are created after provisional application of the plate with two screw holes on either side of the osteotomy. The oblique osteotomy cuts are made through 70% of the bone diameter and the actual osteotomy width is measured. After provisional plate application and removal, the cuts are completed and the plate is re-applied. Eccentric drilling of up to five holes of a 6-hole plate using the 3.5 mm universal drill guide allows closure and compression of osteotomy gaps of up to 4 mm. An interfragmentary screw is placed across the oblique osteotomy through the remaining hole. The technique is simple but requires careful planning and execution. PMID- 15336749 TI - A controlled clinical trial of postoperative hand elevation at home following day case surgery. AB - Although elevation of the upper limb is considered valuable for the prevention and of the reduction of swelling following major surgery or severe injuries to the hand, it is not clear how much elevation, if any, is required following minor surgery such as carpal tunnel decompression. We investigated this by randomizing patients undergoing carpal tunnel decompression into two groups - one having high elevation at home and one being treated with a simple sling. Volumetric analysis of the swelling of the hand 5 days postoperatively showed no significant difference between the two groups. In the trial group, the mean increase in volume of the operated hand was 11 ml (95% CI +4 to +17) or 2.7%. In the control group, the mean swelling was 13 ml (95% CI +4 to +21) or 3.6%. The findings of this study do not support the use of routine high arm elevation following day case surgery of the hand. PMID- 15336750 TI - The role of supinator in the pathogenesis of chronic lateral elbow pain: a biomechanical study. AB - The relative contributions of the forearm extensors to the tensile force at the lateral epicondyle were examined by implanting a force transducer in the common extensor tendon of four soft fixed cadaver elbows and sequentially stretching each muscle arising from the lateral epicondye. Extensor carpi radialis brevis and extensor digitorum communis produced the largest increases while the superficial head of supinator produced a moderate increase in tensile force in the common extensor tendon. Extensor carpi radialis longus and extensor carpi ulnaris had no significant effect. Radial tunnel pressure was measured using a balloon catheter in a separate study of five cadaver elbows. Radial tunnel pressure increased on moving the wrist from neutral to a flexion-pronation position. This positional rise in pressure was reduced by supinator musculotendinous lengthening (77%) while lengthening of the extensor carpi radialis brevis and extensor digitorum communis had no effect. This study demonstrates a biomechanical basis for the superficial head of supinator in the aetiology of both lateral epicondylitis and radial tunnel syndrome. PMID- 15336751 TI - CT angiography in complex upper extremity reconstruction. AB - Computed tomography angiography is a new technique that provides high-resolution, three-dimensional vascular imaging as well as excellent bone and soft tissue spatial relationships. The purpose of this study was to examine the use of computed tomography angiography in planning upper extremity reconstruction. Seventeen computed tomography angiograms were obtained in 14 patients over a 20 month period. All studies were obtained on an outpatient basis with contrast administered through a peripheral vein. All the studies demonstrated the pertinent anatomy and the intraoperative findings were as demonstrated in all cases. Information from two studies significantly altered pre-operative planning. The average charge for computed tomography angiography was 1,140 dollars, compared to 3,900 dollars for traditional angiography. PMID- 15336752 TI - Surgical treatment of major vascular malformations localized to the flexor digitorum superficialis muscle: report of 3 cases. AB - Three cases of congenital arteriovenous malformations localized to the flexor digitorum superficialis presented to a single surgeon. In all three cases, the tumours were resectable with negligible morbidity to the upper extremity. PMID- 15336753 TI - The distal radial fracture in elderly women and the bone mineral density of the lumbar spine and hip. AB - The incidence of distal radial fractures in elderly women is high and is associated with osteoporosis and hip fracture. Osteoporosis can be detected by measuring the bone mineral density (BMD) of the lumbar spine or hip with dual energy X-ray absorptiometry. Low BMD of the lumbar spine or hip is a strong predictor for future vertebral deformities and hip fractures. At present, elderly women with a distal radial fracture are not investigated for osteoporosis on a routine basis. The BMD of the lumbar spine and hip were assessed in 94 women (mean age, 69 years) with a distal radial fracture. A low BMD was found in 85% of the patients, and osteoporosis was diagnosed in 51%. The mean BMD decreased by 0.04 SD per year and there was a significant relationship between post-menopausal status and decreased BMD of the hip. The BMD in patients treated with bisphosphonate medication increased significantly in 1 year. As more than half of the elderly women with a distal radial fracture have osteoporotic BMD values for the lumbar spine or hip, it is our opinion that such patients should be screened for osteoporosis. PMID- 15336754 TI - The Patient Outcomes of Surgery-Hand/Arm (POS-Hand/Arm): a new patient-based outcome measure. AB - The purpose of this study was to develop and validate a new patient-based outcome measure for hand/arm disorders for use in audit, clinical trials and effectiveness studies. There were three stages. First, we carried out interviews with 40 patients with hand/arm disorders to develop and pilot questionnaire content. Second, in a postal survey with 165 pre- and 181 post-surgery patients, we reduced the number of items and identified scales. Third, in a postal survey with 132 pre- and 204 post-surgery patients we evaluated the psychometric properties of the measure. Findings confirmed the acceptability, reliability, validity and responsiveness to clinical change of the questionnaire. The Patient Outcomes of Surgery-Hand/Arm (POS-Hand/Arm) is a new surgical outcome measure that can be used before and after surgery (29 and 33 items, respectively) to evaluate and compare new techniques, surgical teams and units. PMID- 15336755 TI - Chondrocalcinosis of the wrist. AB - Calcium Pyrophosphate Dihydrate Deposition (CPDD) disease has characteristic radiographic features including soft tissue calcification, joint space narrowing, bone sclerosis, subchondral cyst formation without osteophyte formation, and large intraosseous geodes. Triangular fibrocartilage calcification is frequently found and isolated scapho-trapezio-trapezoid (STT) arthritis is specific for CPDD. Distal radio-ulnar (DRUJ), isolated midcarpal joint and piso-triquetral joint involvement also occur. 127 patients were reviewed. Seventy-eight had symptomatic STT joint arthritis, for which 36 underwent surgery. Twenty-two patients had a SLAC wrist deformity for which ten underwent surgery. Eight patients had isolated midcarpal arthritis for which three midcarpal arthrodeses, two four-bone arthrodeses and two carpal tunnel releases were performed. Nineteen patients had a generalized arthritis and seven of the patients underwent surgery: four-corner arthrodesis+scaphoidectomy (one case), carpal tunnel release (two cases) extensor synovectomy (two cases) and trigger finger release (two cases). PMID- 15336756 TI - Chronic finger joint instability reconstructed with bone-ligament-bone graft from the iliac crest. AB - Five patients with chronic instability of digital joints presented with instability and functional disability. Two patients had ulnar collateral ligament damage of the thumb metacarpophalangeal joint and another had chronic multidirectional instability due to radial collateral ligament, dorsal capsule and palmar plate laxity of the metacarpophalangeal joint of the thumb. The fourth patient had a lax radial collateral ligament and palmar plate of the proximal interphalangeal joint of the little finger and the fifth had chronic laxity of the ulnar collateral ligament of the interphalangeal joint of the thumb. All were reconstructed with bone-ligament-bone graft harvested from the iliac crest. The graft was fixed with screws and joint stability was achieved intra-operatively in all patients. All patients achieved a stable joint with improved functional performance at final follow-up. PMID- 15336757 TI - A1 pulley release of locked trigger digit by percutaneous technique. AB - We performed 118 percutaneous releases of the locked trigger digits in an office setting using a specially designed knife. Thirty-five digits were locked in flexion, 79 digits in extension and the remaining four were fixed in a semiflexed position. Successful percutaneous release was achieved for 107 digits (91%), with the remaining 11 digits requiring an open surgical procedure. Although there were no persistent triggering in 98 digits with a follow-up of at least 6 months, painful stiffness at the interphalangeal joints remained in ten digits despite of physical therapy. No neurovascular injury occurred. We suggest that a locked trigger digit can be successfully released with the percutaneous technique. PMID- 15336758 TI - Dental rolls: a suitable model for practising tendon repair techniques. AB - Surgical training has changed significantly in the United Kingdom in last decade. A simulator for practising various tendon repairing techniques is presented. It has been designed for the use of junior trainees in Hand surgery dealing with the routine hand trauma cases and can be assembled quickly and cheaply. PMID- 15336759 TI - Isolated ischemic contracture of the mobile wad: a report of two cases. AB - Two children with isolated ischaemic contractures of the mobile wad (extensor carpi radialis longus, extensor carpi radialis brevis and brachioradialis), which developed after treatment of a supracondylar fracture, are presented. Z lengthening of the extensor carpi radialis longus and brevis tendons improved wrist flexion. PMID- 15336760 TI - Ischaemic contracture in an infant's forearm--a case report. AB - We present a case of a gradually developing ischaemic contracture of the forearm muscles of an infant who developed without any trauma or acute gangrene at birth. Release of the middle and ring finger digitorum profundus muscles and pronator quadratus at 2 years of age corrected the deformity. Histopathology showed no evidence of fibromatosis or any other tumor. Although a dynamic splint was used to maintain the range of motion, the range of the middle finger motion deteriorated gradually 2 years after surgery. Though the pathogenesis of this problem was unclear, we assume that it was caused by fibrosis of muscles as a result of bleeding before or during delivery. PMID- 15336761 TI - End-to-side nerve suturing technique. PMID- 15336762 TI - Rational approach to diagnosis and treatment of ameloblastomas and odontogenic keratocysts. AB - We present decision trees on the treatment of cystic lesions of the jaws based on their location. We give special consideration to the treatment of potentially aggressive lesions such as odontogenic keratocysts and cystic ameloblastomas. The treatment plan is based on a retrospective study of 19 ameloblastomas and similar published studies combined with a prospective study of keratocysts. PMID- 15336763 TI - Relation between size of odontogenic jaw cysts and the pressure of fluid within. AB - Intracystic fluid pressure may have a crucial role in the growth of odontogenic jaw cysts. In this study, we investigated the relation between the size of the cyst and the pressure of the fluid within odontogenic keratocysts, dentigerous cysts, and radicular cysts. The radiolucent area of the cyst on a panoramic radiograph was linearly related to the volume in the cavity, and the correlation coefficient (gamma) was 0.70 (n = 25, P < 0.001). Intracystic fluid pressure correlated negatively with the radiolucent area in odontogenic keratocysts (gamma = -0.76, n = 9, P = 0.02), dentigerous cysts (gamma = -0.54, n = 16, P = 0.03), and radicular cysts (gamma = -0.69, n = 10, P = 0.03). The values of [(intracystic fluid pressure (mmHg)) x (radiolucent area (cm(2)))] did not differ significantly among the three types of cyst. Intracystic fluid pressure may therefore be negatively related to the size of all three types of cyst. PMID- 15336764 TI - Three-dimensional assessment of facial soft-tissue asymmetry before and after orthognathic surgery. AB - We developed a new three-dimensional (3D) method of analysis of facial asymmetry and applied it to 44 patients: 20 Class III cases treated by bimaxillary osteotomy; 12 Class III cases treated by maxillary advancement alone, and 12 Class II cases treated by bimaxillary operations. 3D images were taken within a week before operation (T1), and one (T2), three (T3), and six (T4) months after operation. Landmarks were digitised on each 3D model and facial asymmetry scores were calculated. In the bimaxillary osteotomy group, facial symmetry improved after operation. In the maxillary advancement group, there was no notable improvement in facial symmetry after operation. In the bimaxillary group, facial symmetry deteriorated after operation, particularly at the tip of the nose and prominence of the chin. At six months follow-up, the changes in facial asymmetry in the three groups were not significant. PMID- 15336765 TI - Intra-articular oxidative state correlated with the pathogenesis of disorders of the temporomandibular joint. AB - We investigated the redox state of albumin in the synovial fluid from patients with temporomandibular joint (TMJ) disorders (TMD) to evaluate the relation between the cause of the TMD and the number of types of oxygen in synovial fluid. The albumin was fractionated into three components, human mercaptalbumin (HMA, reduced form) and two types human non-mercaptalbumin (HNA, oxidized form), by high-performance liquid chromatography (HPLC). The 63 patients were divided into three groups radiologically, and the ratios of the redox state of the synovial fluid in each group were compared. The fraction of HNA was significantly higher in patients with advanced disease than in patients with early disease. This indicates that the TMJ is affected by intra-articlular oxidative stress, and the severity of TMD correlates closely with the number of oxidative factors. Oxidative stress was thought to be responsible for the genesis of TMD. PMID- 15336766 TI - Histological assessment of bioengineered new bone in repairing osteoperiosteal mandibular defects in sheep using recombinant human bone morphogenetic protein-7. AB - Numerous experimental studies have been published about osteoinductive bone morphogenetic proteins (BMPs). However, to our knowledge there has been no detailed histological study of a mandibular defect in a large mammal, reconstructed using BMPs. We describe here the histological features of rhBMP-7 induced bone in mandibular defects in sheep. METHODS: A 35 mm osteoperiosteal defect was created at the parasymphyseal region of the mandible in six adult sheep. The continuity of the mandible was maintained using a bony plate, and rhBMP-7 was applied on a type I collagen carrier. Bone labels were injected at selected time intervals during the follow-up period. The animals were killed after 3 months and bone samples were examined histologically, histomorphometrically, and by fluorescence microscopy. RESULTS AND CONCLUSIONS: We found a mixture of woven and lamellar bone that contained many cells with large nuclei. This had not reorganised to form cortical bone and the rhBMP-7 induced bone was more porous than the native bone. The newly-formed bone restored both endosteal and periosteal layers. rhBMP-7-induced bone was biocompatible and induced no ossification of soft tissue or abnormal growth of nearby vital structures. The mineral apposition rate was 1.98 microm/day (range 0.62-5.63 microm/day), a value close to that reported in humans. This suggests that BMPs have a limited effect in accelerating the rate of mineralisation, but promote the pre-mineralisation processes, and perhaps the formation of woven bone. PMID- 15336767 TI - Patients' experiences after undergoing orthognathic surgery at NHS hospitals in the south west of England. AB - Patients are being increasingly involved in assessing the quality of care that they receive in the NHS. Our aim was to undertake a retrospective postal survey to evaluate their perception of the delivery of orthognathic surgery in the south west of the United Kingdom (UK) using a patient-centred measure. A total of 327 patients (53% response rate) participated. Although most participants (n = 249, 76%) reported that they were well-informed about what to expect during treatment, many reported that the symptoms of pain, swelling, or difficulty in eating that they experienced immediately post-operatively were worse than expected. A third also reported that it took them longer to recover from the operation than they had anticipated. Patients undergoing orthognathic surgery in the south west of the UK need more specific information about what to expect both immediately post operatively and at home after discharge. PMID- 15336768 TI - Comparison of palatal sensitivity after treatment of cleft palate by a supraperiosteal or mucoperiosteal flap. AB - We compared palatal sensitivity after push-back palatoplasty in patients operated on with a conventional mucoperiosteal flap and with a supraperiosteal flap. We studied 37 patients, whose mean age was 18 years (range 11-28), with unilateral cleft lip, alveolus, and palate who required palatoplasty at our clinic; 18 patients had a supraperiosteal, and 19 a mucoperiosteal flap. Thresholds of palatal sensitivity were measured with Semmes-Weinstein monofilaments. The mean values of thresholds of palatal sensitivity in patients who had a push-back palatoplasty were higher than those in a normal control group. The mean values for patients who had a supraperiosteal flap were significantly lower than those who had a mucoperiosteal flap. PMID- 15336769 TI - Cartilaginous choristoma deep in the upper midline oral vestibule. AB - The choristoma is a tumour-like mass of normal cells in an abnormal location. Intraoral cartilaginous choristoma is a rare lesion. We report on a cartilaginous choristoma arising in the upper midline oral vestibule. PMID- 15336770 TI - Sustained release of doxycycline for the treatment of peri-implantitis: randomised controlled trial. AB - With the increased use of osseointegrated implants and with many implants functioning for a long time, the treatment of peri-implantitis has become important. Animal studies and clinical case reports have shown that the principle of guided bone regeneration can be applied to the surgical treatment of moderate to profound loss of bone around the implant, but we have found no published clinical studies. PATIENTS AND METHODS: Twenty-eight patients whose ages ranged from 25 to 78 years and who had a total of 48 peri-implant defects were examined at baseline (week 0) and after 18 weeks. This included the recording of bleeding on probing, pocket probing depths, and probing attachment levels at six sites for each tooth. For 2-18 weeks before week 0 all patients had been treated for peri implantitis, including motivation, instruction in oral hygiene, and implant scaling with a hand plastic instrument. They were then randomly allocated to continue with this treatment or to have in addition mechanical debridement and local application of Atridox trade mark which slowly release doxycycline. RESULTS: Patients treated with Atridox trade mark showed a significantly greater gain in mean (S.D.) probing attachment levels than those not treated with Atridox. Only subjects treated with Atridox had a significant gain in mean bleeding on probing (P = 0.001). Application of the biodegradable sustained release device after initial periodontal treatment resulted in a significant gain in mean probing attachment levels in the Atridox trade mark group and a significant reduction in pocket probing depths. There was also a significant difference in mean probing attachment levels (0.6mm). PMID- 15336771 TI - Pedicled sandwich plasty: a variation on alveolar distraction for vertical augmentation of the atrophic mandible. AB - We describe a method of vertical augmentation of an edentulous mandible that causes minimal weakening of bone and disturbance of sensation. PMID- 15336772 TI - Acquired syngnathia. AB - We report two cases of acquired syngnathia, in case 1 probably as a result of a fibrotic tubed pedicle and in case 2 from myositis ossificans. The treatment plan for syngnathia must include not only operation but also a rehabilitation programme (particularly muscle training) PMID- 15336773 TI - Piezosurgery: an ultrasound device for cutting bone and its use and limitations in maxillofacial surgery. AB - Piezosurgery uses modulated ultrasonic vibration to allow controlled cutting of bony structures. Delicate bony structures can be cut easily and with great precision, without destruction of soft tissue. We have found this device useful when exact cutting of thin bones is essential. However, it is of only limited use in cutting thick bones and in regions with limited access. PMID- 15336774 TI - Treatment of severe peri-implant bone loss using autogenous bone and a bioabsorbable polymer that delivered doxycycline (Atridox). AB - With an increase of the number of implants being inserted, it is inevitable that the number of cases of peri-implantitis with loss of bone will rise. We report a case in which an autogenous cancellous bone graft was placed into the peri implant bone defect and given protection with a bioabsorbable polymer barrier (Atridox) that released doxycycline slowly. PMID- 15336775 TI - Solitary mucosal neuroma of the hard palate: a case report. AB - Mucosal neuroma is rare neurogenic tumour that arises in the oral cavity as a component of multiple endocrine neoplasia type 2b (MEN 2b). We present the case of a histologically confirmed solitary mucosal neuroma of the hard palate in a 49 year-old woman with no other components of MEN 2b. PMID- 15336776 TI - Old habits die hard. PMID- 15336777 TI - Historical case: Wilhelm Fredrich Von Ludwig (1790-1865). PMID- 15336778 TI - HLA alleles, IFN-gamma responses to HPV-11 E6, and disease severity in patients with recurrent respiratory papillomatosis. AB - Recurrent respiratory papillomatosis (RRP) remains an immunologic enigma. Human papillomavirus (HPV) types 6 and 11 are the predominant HPV viruses that cause papilloma development. However, it is unclear why only a very small fraction of HPV-exposed individuals develop RRP. We performed high-resolution HLA class I and II genotyping on 70 randomly selected patients (56 Caucasians and 14 African Americans) with RRP. We report, for the first time, an increased frequency of HLA DRB1*0102 in Caucasian patients with RRP, suggesting that this allele predisposes individuals to RRP. Additionally, HLA-DRB1*0301, DQB1*0201, and DQB1*0202 alleles were selectively enriched in Caucasians with severe disease, suggesting that these alleles may regulate disease severity. In contrast, HLA-DQB1*0602 was more frequent in controls than in Caucasians with severe disease, suggesting a severity-sparing effect of this allele. Furthermore, both DQB1*0201 and DQB1*0202 were enriched, whereas DQB1*0602 was absent, in African-Americans. Interestingly, HLA-DRB1*0301 and DQB1*0201 correlated with reduced interferon-gamma expression in patients with RRP. Larger studies are needed to identify other class II major histocompatibility complex alleles that may influence disease predisposition, disease severity, or both, especially in African-American patients, to ultimately illuminate the regulatory effects of these alleles in the predisposition and severity of RRP. PMID- 15336779 TI - Is TAP2*0102 allele involved in insulin-dependent diabetes mellitus (type 1) protection? AB - In this study, we have investigated the frequencies of TAP1 and TAP2 alleles in a group of 226 persons, living in La Reunion Island, consisting of 70 patients with insulin-dependent diabetes mellitus (IDDM) and most of their first degree relatives (i.e., 156 parents and full sibling subjects) and previously HLA DQB1, DQA1, and DRB1 genotyped. The population of this island is constituted by a particular structure of highly crossbreeding people. Interestingly, the new TAP2*0104 allele, previously discovered by our team in Reunion Island, was found to be increased in the IDDM population and the calculated HRR was relatively high (HRR = 3.3). This result seems to be due to a positive linkage disequilibrium between TAP2*0104 allele and the highly diabetogenous DQB1* 0201-DQA1* 0501-DRB1 0301 haplotype (HRR = 9), which suggests that TAP2*0104 cannot be considered as an additional predispositional factor, but more as a genetic susceptibility marker of IDDM. In addition, we show that minor alleles (TAP2D, *0102, *0103, *0104) are associated with a restricted number of HLA DQ-DR haplotypes and each of them exhibits a preferential linkage with one particular haplotype. In contrast with other alleles, and despite a HRR value close to 1, we show that TAP2*0102 allele contributes significantly to a drastic reduction of the diabetogenic effect of DQB1*0201-DQA1*0301.1-DRB*0701 haplotype. Indeed, this haplotype, which is usually preferentially transmitted to affected children, is dominantly transmitted to healthy children when it is associated with TAP2*0102. Therefore, this allele seems to contribute to genetic protection to IDDM. PMID- 15336780 TI - Regulatory T-cell response and tumor vaccine-induced cytotoxic T lymphocytes in human melanoma. AB - A role of CD4(+) cells in the regulation of immune responses has steadily gained renewed recognition. The understanding of these T-regulatory (T-reg) cells in the generation of antitumor cytolytic T lymphocyte (CTL) response is therefore important. It has been shown that immunization with specific peptides, DNA, or tumor lysate-based vaccines can induce CTL responses in vivo. We have immunized melanoma patients with major histocompatibility complex (MHC) class I restricted peptide- or melanoma tumor lysate-loaded antigen-presenting cell (APC)-based vaccines and have monitored the generation of CTL responses and T-reg cell responses, if any. Using tetramer staining and limiting dilution analyses as monitors of CTL responses, we found significant increases in the number of antigen-specific CTL in circulation after vaccination with the MART-1(27-35) peptide (AAGIGILTV)-pulsed autologous APC, the MAGE-1(161-169) peptide (EADPTGHSY)-pulsed APC, or with autologous tumor lysate-pulsed APC. The antigen specific CTL reached the peak expansion by day 7 and then declined to the prevaccine levels by day 28. The decline in the CTL response was associated by a concomitant expansion of CD4(+) CD25(+)T cells. Analysis of postvaccine peripheral blood lymphocytes (PBL) from patients showed an increased amount of interleukin (IL)-10 secretion on in vitro stimulation with IL-2 after successive vaccination. Triple color flow cytometric analyses revealed cytoplasmic IL-10 in the CD4(+)CD25(+) T-cell fraction and the number of CD4(+)CD25(+) IL-10(+) T cells were found to increase significantly in postvaccine PBL. These observations have implications in tumor antigen and APC/dendritic cell (DC)-based cancer vaccine strategies. PMID- 15336781 TI - Pronase treatment facilitates alloantibody flow cytometric and cytotoxic crossmatching in the presence of rituximab. AB - Rituximab (RIT), a murine/human chimeric monoclonal antibody directed against human CD20 is under investigation for its role in transplantation. RIT causes B cell crossmatches to appear positive. Pronase, a proteolytic enzyme that targets F(c) receptors removes CD20 from B cells. After CD20 is removed, RIT should not bind, making it possible to detect class I or class II antibodies on treated B cells. In this study, we incubated RIT with normal human serum (NHS, negative control) or pooled sera from highly sensitized (>50% panel reactive antibody, HLA+) subjects awaiting renal transplantation (positive control) and then performed B-cell flow cytometric crossmatches using untreated or pronase treated B cells as targets. We observed that untreated B cells incubated with RIT-spiked NHS displayed a significant increase in surface fluorescence compared with NHS without RIT, similar to the fluorescence that occurs with a positive crossmatch. In contrast, when CD20 was cleaved from the B cells with pronase, B cells displayed a negative crossmatch with the RIT-spiked NHS. In addition, there was no change in the crossmatches of pooled high panel reactive antibody (PRA) sera after pronase treatment. RIT could be used without worry about losing the ability to perform transplant immunologic monitoring. PMID- 15336782 TI - Good kidney transplant outcome in recipients with presensitization against HLA class II but not HLA class I. AB - It is a matter of debate whether pretransplant anti-human leukocyte antigen (HLA) class II antibodies contribute to the increased graft rejection rate found in presensitized recipients. We investigated the influence of preformed anti-HLA class II antibodies on graft survival in 5949 cadaver kidney transplants. Pretransplant recipient sera were tested in enzyme-linked immunosorbent assay (ELISA) for immunoglobulin (Ig)G-anti-HLA class I and IgG-anti-HLA class II antibodies. A total of 672 recipients with antibodies against HLA class II but not against HLA class I had a 3-year graft survival rate of 80 +/- 2%, identical to the 80 +/- 1% rate in 4561 recipients who were negative for anti-HLA class I and II (p = NS). Graft survival was significantly lower in 365 recipients who were positive for both anti-HLA class I and II (65 +/- 3%, p < 0.0001). Compatibility for HLA-A+B+DR influenced graft survival significantly in anti-HLA class I- and II-positive recipients (p = 0.0016), whereas no significant HLA effect was found in patients with antibodies directed against only class I or II. Surprisingly, not even incompatibility for HLA class II antigens of the DR locus caused a significant impairment of graft survival in anti-class II-positive recipients. We conclude that the risk associated with sensitization against HLA class II in the absence of sensitization against HLA class I is negligible. PMID- 15336783 TI - Allogeneic core amino acids of an immunodominant allopeptide are important for MHC binding and TCR recognition. AB - The indirect alloimmune response seems to be restricted to a few dominant major histocompatibility complex (MHC)-derived peptides responsible for T-cell activation in allograft rejection. The molecular mechanisms of indirect T-cell activation have been studied using peptide analogues derived from the dominant allopeptide in vitro, whereas the in vivo effects of peptide analogues have not been well characterized yet. In the present study, we generated allochimeric peptide analogues by replacing the three allogeneic amino acids 5L, 9L, and 10T in the sequence of the dominant MHC class I allopeptide P1. These allochimeric peptide analogues were used to define the allogeneic amino acids critical for the MHC binding and TCR recognition. We found that position 5 (5L) of the dominant allopeptide acts as an MHC-binding residue, while the other two allogeneic positions, 9 and 10, are important for the T-cell receptor (TCR) recognition. A peptide containing the MHC-binding residue 5L, as the only different amino acid between donor (RT1.A(u)) and recipient (RT1.A(l)) sequences, did not induce proliferation of lymph node cells primed with the dominant peptide and prevented dominant peptide-induced acceleration of allograft rejection. Identification of MHC and TCR contact residues should facilitate the development of antigen specific therapies to inhibit or regulate the indirect alloimmune response. PMID- 15336784 TI - HLA-B27 genotyping by fluorescent resonance emission transfer (FRET) probes in real-time PCR. AB - Several polymerase chain reaction (PCR)-based human leukocyte antigen (HLA) genotyping methods are in use, but none is fully satisfactory. The introduction of real-time PCR (rt-PCR) with fluorescence resonance energy transfer (FRET) probes provides a powerful tool to overcome the drawbacks of current methods such as the long processing time and the requirement for post-PCR manual procedures. Here we present evidence that the FRET-fluorotyping principle may resolve HLA-B27 variants, providing a higher resolution in less time than the techniques currently in use. The protocol uses between one and three consecutive amplification reactions depending on the resolution required. The first reaction, aimed at detecting HLA-B27-positive samples, uses beta-globin coamplification as control. The second reaction, aimed at resolving most frequent B27 alleles, uses two hybridization probes whose melting temperatures curves allow the classification of HLA-B27 alleles into eight groups. By adding a third reaction, even the rarest alleles associated and not associated to ankylosing spondylitis (AS) may be discriminated. The technique was blindly tested on 60 samples from individuals previously typed and confirmed by standard PCR sequence-specific oligoprobes-PCR sequence and PCR-based typing PCR-SBT (30 B27+, 30 non-B27). There was a complete concordance rate, thus confirming the potential of this new technique for clinical HLA-B27 typing and for HLA typing in general. PMID- 15336785 TI - Association of a novel single nucleotide polymorphism in the human perforin gene with the outcome of HTLV-I infection in patients from northeast Iran (Mash-had). AB - Human T lymphotropic virus I (HTLV-I)-specific cytotoxic T lymphocytes (CTL) recognize the products of the HTLV-I Tax, in the context of HLA-A2 and kill their target through a perforin-dependent mechanism. The efficiency of the CTL response may lead HTLV-I-infected individuals to remain carriers or to the development of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Perforin is a cytolytic molecule that contributes to CTL-mediated killing of virus-infected cells. Thus polymorphism in the perforin gene may determine the efficiency of the CTL response in HTLV-I-infected individuals. In this study, we performed single stranded conformational polymorphism (SSCP) and DNA sequencing to analyze the promoter, 5' UTR and first intron of the perforin gene to identify novel polymorphisms. We detected a novel polymorphism in the first intron at position +418*C/T, relative to the transcription start site. Genotyping of patients with HAM/TSP, HTLV-I carriers, and healthy controls revealed that the frequency of the C allele was statistically significantly increased in HAM/TSP patients compared with healthy controls group (p = 0.005). The frequency of the C allele was higher, but not significantly so, in the HAM/TSP group compared with HTLV-I carriers (p = 0.09), whereas there was no difference between HTLV-I carriers and healthy controls. Our results suggest that the perforin +418*C/T polymorphism is associated with the outcome of HTLV-I infection. PMID- 15336786 TI - HLA DRB1* and DQB1* allelic diversity among Nadars: a primitive south Indian Dravidian caste group. AB - South India is one of the oldest geophysical regions occupied mainly by Dravidian language-speaking people. In the present study, 84 unrelated healthy Nadar individuals from Tamil Nadu State were analyzed for human leukocyte antigen (HLA) DRB1* and DQB1* allele frequencies and their haplotype frequencies were analyzed by high resolution polymerase chain reaction (PCR)-typing of genomic DNA. Results revealed that Nadars possessed >0.1 allele frequencies of HLA DRB1*15011, 03, 04, 1404, 0701, 1001, and DQB1*0201, 03, 0501, and 0503. The highest frequencies of DRB1*0317 (0.048), DRB1*1317 (0.012), and DRB1*1404 (0.107) in the world were found among the Nadar caste. The most frequent two-locus haplotypes identified among the Nadars were DRB1*0701-DQB1*0201 and DRB1*1000-DQB1*0501, and DRB1*1404 DQB1*0503 and DRB1*15011-DQB1*0601. When compared with world populations, HLA DRB1*1322, DRB1*1434, and DQB1*0614 were identified exclusively among Nadars. The heterozygosity and Watterson's homozygosity tests indicated that balancing selection still operates among the South Indian caste groups. The hierarchical cluster analysis for DRB1 data from the Indian caste and population groups showed that the Nadars cluster with Kashmir Brahmins and North Indians. The present study shows that the Nadar caste has several unique alleles and haplotypes at high frequency that are rare or absent in other populations of India and the world. PMID- 15336787 TI - Impact of transverse and longitudinal dispersion on first-order degradation rate constant estimation. AB - A two-dimensional analytical model is employed for estimating the first-order degradation rate constant of hydrophobic organic compounds (HOCs) in contaminated groundwater under steady-state conditions. The model may utilize all aqueous concentration data collected downgradient of a source area, but does not require that any data be collected along the plume centerline. Using a least squares fit of the model to aqueous concentrations measured in monitoring wells, degradation rate constants were estimated at a former manufactured gas plant (FMGP) site in the Midwest U.S. The estimated degradation rate constants are 0.0014, 0.0034, 0.0031, 0.0019, and 0.0053 day(-1) for acenaphthene, naphthalene, benzene, ethylbenzene, and toluene, respectively. These estimated rate constants were as low as one-half those estimated with the one-dimensional (centerline) approach of Buscheck and Alcantar [Buscheck, T.E., Alcantar, C.M., 1995. Regression techniques and analytical solutions to demonstrate intrinsic bioremediation. In: Hinchee, R.E., Wilson, J.T., Downey, D.C. (Eds.), Intrinsic Bioremediation, Battelle Press, Columbus, OH, pp. 109-116] which does not account for transverse dispersivity. Varying the transverse and longitudinal dispersivity values over one order of magnitude for toluene data obtained from the FMGP site resulted in nearly a threefold variation in the estimated degradation rate constant highlighting the importance of reliable estimates of the dispersion coefficients for obtaining reasonable estimates of the degradation rate constants. These results have significant implications for decision making and site management where overestimation of a degradation rate may result in remediation times and bioconversion factors that exceed expectations. For a complex source area or non steady-state plume, a superposition of analytical models that incorporate longitudinal and transverse dispersion and time may be used at sites where the centerline method would not be applicable. PMID- 15336788 TI - Magnetic resonance imaging of nonaqueous phase liquid during soil vapor extraction in heterogeneous porous media. AB - Soil vapor extraction (SVE) is commonly used to remediate nonaqueous phase liquids (NAPLs) from the vadose zone. This paper aims to determine the effect of grain size heterogeneity on the removal of NAPL in porous media during SVE. Magnetic resonance imaging (MRI) was used to observe and quantify the amount and location of NAPL in flow-through columns filled with silica gel grains. MRI is unique because it is nondestructive, allowing three-dimensional images to be taken of the phases as a function of space and time. Columns were packed with silica gel in three ways: coarse grains (250-550 microm) only, fine grains (32-63 microm) only, and a core of fine grains surrounded by a shell of coarse grains. Columns saturated with water were drained under a constant suction head, contaminated with decane, and then drained to different decane saturations. Each column was then continuously purged with water-saturated nitrogen gas and images were taken intermittently. Results showed that at residual saturation, a sharp volatilization front moved through the columns filled with either coarse-grain or fine-grain silica gel. In the heterogeneous columns, the volatilization front in the core lagged just behind the shell because gas flow was greater through the shell and decane in the core diffused outward to the shell. When decane saturation in the core was above residual saturation, decane volatilization occurred near the inlet, the relative decane saturation throughout the core dropped uniformly, and decane in the core flowed in the liquid phase to the shell to replenish volatilized decane. These results indicate that NAPL trapped in low permeability zones can flow to replenish areas where NAPL is lost due to SVE. However, when residual NAPL saturation is reached, NAPL flow no longer occurs and diffusion limits removal from low-permeability zones. PMID- 15336789 TI - Infiltration of PCE in a system containing spatial wettability variations. AB - A two-dimensional infiltration experiment was conducted to investigate and quantify the effect of spatial wettability variations on DNAPL migration and entrapment in saturated sands. Experimental observations of tetrachloroethylene (PCE) infiltration showed that organic-wet sand lenses acted as very effective capillary barriers, retaining PCE and inhibiting its downward migration. A multiphase numerical simulator was used to model this sand box experiment. The simulator incorporates wettability-modified van Genuchten and Brooks-Corey capillary pressure/saturation relationships as well as Burdine and Mualem relative permeability relationships. PCE mass distributions, estimated by image analysis of digital photographs taken during the infiltration event, were compared to simulation results. Although both relative permeability models were qualitatively able to predict the PCE retention in the organic-wet layers, simulations with the Mualem model failed to capture the observed rate of PCE migration. A traditional multiphase simulator, incorporating water-wet capillary retention relations, failed to predict both PCE pathways and retention behavior. This study illustrates the potential influence of subsurface wettability variations on DNAPL migration and entrapment and supports the use of modified capillary relations in conjunction with the Burdine model in multiphase flow simulators. PMID- 15336790 TI - Plant aided bioremediation in the vadose zone: model development and applications. AB - Phytoremediation has the potential to enhance clean up of land contaminated by various pollutants. A mathematical model that includes a two-fluid phase flow model of water flow as well as a two-region soil model of contaminant reactions was developed and applied to various bioremediation scenarios in the unsaturated zone, especially to plant-aided bioremediation. To investigate model behavior and determine the main parameters and mechanisms that affect bioremediation in unplanted and planted soils, numerical simulations of theoretical scenarios were conducted before applying the model to field data. It is observed from the results that parameters affecting the contaminant concentration in the water phase, such as aqueous solubility, the octanol-water partition coefficient, and organic carbon content of the soil controlled the contaminant fate in the vadose zone. Simulation using the developed model also characterized the fate and transport of the contaminants both in planted and unplanted soils satisfactorily for field applications. Although phytoremediation has the potential for remediation of contaminated soils, results from both modeling and field studies suggested that plants may not always enhance the remediation efficiency when the soil already has a high microbial concentration, when the contaminant bioavailability is low, or when the overall reaction is mass transfer-limited. Therefore, other steps to increase contaminant bioavailability are needed in phytoremediation applications; natural purification mechanisms such as aging, volatilization, and natural bioremediation should be considered to maximize the plant effect and minimize the cost. PMID- 15336791 TI - Mechanisms of electron acceptor utilization: implications for simulating anaerobic biodegradation. AB - Simulation of biodegradation reactions within a reactive transport framework requires information on mechanisms of terminal electron acceptor processes (TEAPs). In initial modeling efforts, TEAPs were approximated as occurring sequentially, with the highest energy-yielding electron acceptors (e.g. oxygen) consumed before those that yield less energy (e.g., sulfate). Within this framework in a steady state plume, sequential electron acceptor utilization would theoretically produce methane at an organic-rich source and Fe(II) further downgradient, resulting in a limited zone of Fe(II) and methane overlap. However, contaminant plumes often display much more extensive zones of overlapping Fe(II) and methane. The extensive overlap could be caused by several abiotic and biotic processes including vertical mixing of byproducts in long-screened monitoring wells, adsorption of Fe(II) onto aquifer solids, or microscale heterogeneity in Fe(III) concentrations. Alternatively, the overlap could be due to simultaneous utilization of terminal electron acceptors. Because biodegradation rates are controlled by TEAPs, evaluating the mechanisms of electron acceptor utilization is critical for improving prediction of contaminant mass losses due to biodegradation. Using BioRedox-MT3DMS, a three-dimensional, multi-species reactive transport code, we simulated the current configurations of a BTEX plume and TEAP zones at a petroleum-contaminated field site in Wisconsin. Simulation results suggest that BTEX mass loss due to biodegradation is greatest under oxygen-reducing conditions, with smaller but similar contributions to mass loss from biodegradation under Fe(III)-reducing, sulfate-reducing, and methanogenic conditions. Results of sensitivity calculations document that BTEX losses due to biodegradation are most sensitive to the age of the plume, while the shape of the BTEX plume is most sensitive to effective porosity and rate constants for biodegradation under Fe(III)-reducing and methanogenic conditions. Using this transport model, we had limited success in simulating overlap of redox products using reasonable ranges of parameters within a strictly sequential electron acceptor utilization framework. Simulation results indicate that overlap of redox products cannot be accurately simulated using the constructed model, suggesting either that Fe(III) reduction and methanogenesis are occurring simultaneously in the source area, or that heterogeneities in Fe(III) concentration and/or mineral type cause the observed overlap. Additional field, experimental, and modeling studies will be needed to address these questions. PMID- 15336792 TI - Geochemical stability of phosphorus solids below septic system infiltration beds. AB - Review of 10 mature septic system plumes in Ontario, revealed that phosphorus (P) attenuation commonly occurred close to the infiltration pipes, resulting in discrete narrow intervals enriched in P by a factor of 2-4 (. MSc thesis, Dept. Earth Sci., University of Waterloo, Waterloo, Ont.; Ground Water 36 (1995) 1000; J. Contam. Hydrol. 33 (1998) 405). Although these attenuation reactions appeared to be sustainable under present conditions, the potential for remobilization of this P mass, should geochemical conditions change, is unknown. To test the stability of these P solids, dynamic flow column tests were carried out using sediments from three of the previously studied sites (Cambridge, Langton and Muskoka) focusing on sediments from the 'High-P' and underlying (Below) zones. Tests were continued for 166-266 pore volumes (PVs), during which time varying degrees of water saturation were maintained. During saturated flow conditions, relatively high concentrations of PO4 were eluted from the Cambridge and Langton High-P zones (up to 4 and 9 mg/l P, respectively), accompanied by elevated concentrations of Fe (up to 1.4 mg/l) and Mn (up to 4 mg/l) and lower values of Eh (<150 mV). The Below zones from Cambridge and Langton, however, maintained lower concentrations of P (generally<2 mg/l), Fe (<0.2 mg/l) and Mn (<1 mg/l) and maintained higher Eh (>250 mV) during saturated flow conditions. During unsaturated flow, P and Fe declined dramatically in the High-P zones (P<1 mg/l, Fe<0.2 mg/l), whereas concentrations remained about the same during saturated and unsaturated flow in the Below zones. This behavior is at least partly attributed to the development of reducing conditions during saturated flow in the High-P zones, leading to reductive dissolution of Fe (III)-P solids present in the sediments. Reducing conditions did not develop in the Below zones apparently because of lower sediment organic carbon (OC) contents (0.03-0.04 wt.%) compared to the High-P zones (0.2-0.65 wt.%). At the Muskoka site, where the sediments were noncalcareous, low values of P (<0.2 mg/l) were maintained in both the High P and Below columns and reducing conditions did not develop. Results indicate the possibility of remobilizing P accumulated below septic system infiltration beds should conditions become more reducing. This could occur if sewage loading patterns change, for example when a seasonal use, lakeshore cottage is converted to a permanent dwelling. PMID- 15336793 TI - A triple-continuum approach for modeling flow and transport processes in fractured rock. AB - This paper presents a triple-continuum conceptual model for simulating flow and transport processes in fractured rock. Field data collected from the unsaturated zone of Yucca Mountain, a repository site of high-level nuclear waste, show a large number of small-scale fractures. The effect of these small fractures has not been considered in previous modeling investigations within the context of a continuum approach. A new triple-continuum model (consisting of matrix, small fracture, and large-fracture continua) has been developed to investigate the effect of these small fractures. This paper derives the model formulation and discusses the basic triple-continuum behavior of flow and transport processes under different conditions, using both analytical solutions and numerical approaches. The simulation results from the site-scale model of the unsaturated zone of Yucca Mountain indicate that these small fractures may have an important effect on radionuclide transport within the mountain. PMID- 15336794 TI - Biogeochemical processes at the fringe of a landfill leachate pollution plume: potential for dissolved organic carbon, Fe(II), Mn(II), NH4, and CH4 oxidation. AB - Various redox reactions may occur at the fringe of a landfill leachate plume, involving oxidation of dissolved organic carbon (DOC), CH4, Fe(II), Mn(II), and NH4 from leachate and reduction of O2, NO3 and SO4 from pristine groundwater. Knowledge on the relevance of these processes is essential for the simulation and evaluation of natural attenuation (NA) of pollution plumes. The occurrence of such biogeochemical processes was investigated at the top fringe of a landfill leachate plume (Banisveld, the Netherlands). Hydrochemical depth profiles of the top fringe were captured via installation of a series of multi-level samplers at 18, 39 and 58 m downstream from the landfill. Ten-centimeter vertical resolution was necessary to study NA within a fringe as thin as 0.5 m. Bromide appeared an equally well-conservative tracer as chloride to calculate dilution of landfill leachate, and its ratio to chloride was high compared to other possible sources of salt in groundwater. The plume fringe rose steadily from a depth of around 5 m towards the surface with a few meters in the period 1998-2003. The plume uplift may be caused by enhanced exfiltration to a brook downstream from the landfill, due to increased precipitation over this period and an artificial lowering of the water level of the brook. This rise invoked cation exchange including proton buffering, and triggered degassing of methane. The hydrochemical depth profile was simulated in a 1D vertical reactive transport model using PHREEQC-2. Optimization using the nonlinear optimization program PEST brought forward that solid organic carbon and not clay minerals controlled retardation of cations. Cation exchange resulted in spatial separation of Fe(II), Mn(II) and NH4 fronts from the fringe, and thereby prevented possible oxidation of these secondary redox species. Degradation of DOC may happen in the fringe zone. Re-dissolution of methane escaped from the plume and subsequent oxidation is an explanation for absence of previously present nitrate and anaerobic conditions in pristine groundwater above the plume. Stable carbon isotope (delta13C) values of methane confirm anaerobic methane oxidation immediately below the fringe zone, presumably coupled to reduction of sulfate, desorbed from iron oxide. Methane must be the principle reductant consuming soluble electron-acceptors in pristine groundwater, thereby limiting NA for other solutes including organic micro-pollutants at the fringe of this landfill leachate plume. PMID- 15336795 TI - Transport and reduction of nitrate in clayey till underneath forest and arable land. AB - Transport and reduction of nitrate in a typically macroporous clayey till were examined at variable flow rate and nitrate flux. The experiments were carried out using saturated, large diameter (0.5 m), undisturbed soil columns (LUC), from a forest and nearby agricultural sites. Transport of nitrate was controlled by flow along the macropores (fractures and biopores) in the columns. Nitrate reduction (denitrification) determined under active flow mainly followed first order reactions with half-lives (t(1/2)) increasing with depth (1.5-3.5 m) from 7 to 35 days at the forest site and 1-7 h at the agricultural site. Nitrate reduction was likely due to microbial degradation of accumulated organic matter coupled with successive consumption of O2 and NO3- in the macropore water followed by reductive dissolution of Fe and Mn from minerals along the macropores. Concentrations of total organic carbon measured in soil samples were near identical at the two study sites and consequently not useful as indicator for the observed differences in nitrate reduction. Instead the high reduction rates at the agricultural site were positively correlated with elevated concentration of water-soluble organic carbon and nitrate-removing bacteria relative to the forest site. After high concentrations of water-soluble organic carbon in the columns from the agricultural site were leached they lost their elevated reduction rates, which, however, was successfully re-established by infiltration of new reactive organics represented by pesticides. Simulations using a calibrated discrete fracture matrix diffusion (DFMD) model could reasonably reproduce the denitrification and resulting flux of nitrate observed during variable flow rate from the columns. PMID- 15336796 TI - Interpretation of injection-withdrawal tracer experiments conducted between two wells in a large single fracture. AB - Tracer experiments conducted using a flow field established by injecting water into one borehole and withdrawing water from another are often used to establish connections and investigate dispersion in fractured rock. As a result of uncertainty in the uniqueness of existing models used for interpretation, this method has not been widely used to investigate more general transport processes including matrix diffusion or advective solute exchange between mobile and immobile zones of fluid. To explore the utility of the injection-withdrawal method as a general investigative tool and with the intent to resolve the transport processes in a discrete fracture, two tracer experiments were conducted using the injection-withdrawal configuration. The experiments were conducted in a fracture which has a large aperture (>500 microm) and horizontally pervades a dolostone formation. One experiment was conducted in the direction of the hydraulic gradient and the other in the direction opposite to the natural gradient. Two tracers having significantly different values of the free-water diffusion coefficient were used. To interpret the experiments, a hybrid numerical analytical model was developed which accounts for the arcuate shape of the flow field, advection-dispersion in the fracture, diffusion into the matrix adjacent to the fracture, and the presence of natural flow in the fracture. The model was verified by comparison to a fully analytical solution and to a well-known finite element model. Interpretation of the tracer experiments showed that when only one tracer, advection-dispersion, and matrix diffusion are considered, non-unique results were obtained. However, by using multiple tracers and by accounting for the presence of natural flow in the fracture, unique interpretations were obtained in which a single value of matrix porosity was estimated from the results of both experiments. The estimate of porosity agrees well with independent measurements of porosity obtained from core samples. This suggests that: (i) the injection-withdrawal method is a viable tool for the investigation of general transport processes provided all relevant experimental conditions are considered and multiple conservative tracers are used; and (ii) for the conditions of the experiments conducted in this study, the dominant mechanism for exchange of solute between the fracture and surrounding medium is matrix diffusion. PMID- 15336797 TI - A PCE groundwater plume discharging to a river: influence of the streambed and near-river zone on contaminant distributions. AB - An investigation of a tetrachloroethene (PCE) groundwater plume originating at a dry cleaning facility on a sand aquifer and discharging to a river showed that the near-river zone strongly modified the distribution, concentration, and composition of the plume prior to discharging into the surface water. The plume, streambed concentration, and hydrogeology were extensively characterized using the Waterloo profiler, mini-profiler, conventional and driveable multilevel samplers (MLS), Ground Penetrating Radar (GPR) surveys, streambed temperature mapping (to identify discharge zones), drivepoint piezometers, and soil coring and testing. The plume observed in the shallow streambed deposits was significantly different from what would have been predicted based on the characteristics of the upgradient plume. Spatial and temporal variations in the plume entering the near-river zone contributed to the complex contaminant distribution observed in the streambed where concentrations varied by factors of 100 to 5000 over lateral distances of less than 1 to 3.5 m. Low hydraulic conductivity semi-confining deposits and geological heterogeneities at depth below the streambed controlled the pattern of groundwater discharge through the streambed and influenced where the plume discharged into the river (even causing the plume to spread out over the full width of the streambed at some locations). The most important effect of the near-river zone on the plume was the extensive anaerobic biodegradation that occurred in the top 2.5 m of the streambed, even though essentially no biodegradation of the PCE plume was observed in the upgradient aquifer. Approximately 54% of the area of the plume in the streambed consisted solely of PCE transformation products, primarily cis-1,2-dichloroethene (cDCE) and vinyl chloride (VC). High concentrations in the interstitial water of the streambed did not correspond to high groundwater-discharge zones, but instead occurred in low discharge zones and are likely sorbed or retarded remnants of past high-concentration plume discharges. The high-concentration areas (up to 5529 microg/l of total volatile organics) in the streambed are of ecological concern and represent potential adverse exposure locations for benthic and hyporheic zone aquatic life, but the effect of these exposures on the overall health of the river has yet to be determined. Even if the upgradient source of PCE is remediated and additional PCE is prevented from reaching the streambed, the high-concentration deposits in the streambed will likely take decades to hundreds of years to flush completely clean under natural conditions because these areas have low vertical groundwater flow velocities and high retardation factors. Despite high concentrations of contaminants in the streambed, PCE was detected in the surface water only rarely due to rapid dilution in the river and no cDCE or VC was detected. Neither the sampling of surface water nor the sampling of the groundwater from the aquifer immediately adjacent to the river gave an accurate indication of the high concentrations of PCE biodegradation products present in the streambed. Sampling of the interstitial water of the shallow streambed deposits is necessary to accurately characterize the nature of plumes discharging to rivers. PMID- 15336798 TI - History of the evolution of echocardiography. AB - Echocardiography provides information regarding cardiac morphology, function and hemodynamics non-invasively. It is the most frequently performed cardiovascular examination after electrocardiography and chest X-ray. In less than half a century, this technique has evolved to a mainstay of cardiovascular medicine. The historical evolution of echocardiography is succinctly described including that of M-mode, two-dimensional, Doppler, stress, transesophageal, intraoperative, contrast, digital, three-dimensional and intracardiac echocardiography. PMID- 15336799 TI - Acute ventricular rate control in atrial fibrillation and atrial flutter. AB - Atrioventricular node blocking agents including beta-adrenergic blockers, non dihydropyridine calcium channel blockers and digoxin are usually effective in controlling ventricular rate in atrial fibrillation and flutter. Intravenous beta blockers and non-dihydropyridine calcium channel blockers are equally effective in rapidly controlling the ventricular rate. The addition of digoxin to the regimen causes a favorable outcome but digoxin as a single agent is generally less effective in slowing the ventricular rate in acute setting. Clonidine, magnesium, and amiodarone have also been used for acute ventricular rate control in atrial fibrillation. Limited data suggest that combination regimens provide better ventricular rate control than any agent alone. The agent of first choice is usually individualized depending upon the clinical situation. Beta-blockers are preferable in patients with myocardial ischemia, myocardial infarction and hyperthyroidism and in post-operative state, but should be avoided in patients with bronchial asthma and chronic obstructive pulmonary disease where non dihydropyridine calcium channel blockers are preferred. Beta-blockers are preferred drugs used for acute ventricular rate control in atrial fibrillation during pregnancy. In atrial fibrillation with Wolff-Parkinson-White syndrome, beta-blockers, calcium channel blockers and digoxin should be avoided, as these drugs are selective atrioventricular node blockers without slowing conduction through the accessory pathway, which can lead to increased transmission of impulses preferentially through the accessory pathway and precipitate ventricular fibrillation. The drug of choice for atrial fibrillation in pre-excitation syndrome is procainamide but propafenone, flecainide and disopyramide have also been used. When clinical condition is unstable or patient is hemodynamically compromised, immediate electrical cardioversion is the treatment of choice, as the best measure to control ventricular rate is by conversion to sinus rhythm. Factors precipitating rapid ventricular rate should be treated as well. PMID- 15336800 TI - Wasting of the left ventricle in patients with cardiac cachexia: a cardiovascular magnetic resonance study. AB - BACKGROUND: The "cachectic heart" has been described as a pathologic decrease in the size and mass of the heart, but no in vivo studies have shown changes in cardiac dimensions or left ventricular (LV) mass over time in chronic heart failure (CHF) associated with body wasting (cardiac cachexia). Cardiovascular magnetic resonance (CMR) has high reproducibility and is more sensitive than other techniques. METHODS: CMR studies of LV volumes and mass were performed at baseline and a mean of 15 months later in nine CHF patients with cardiac cachexia and 28 matched CHF controls without cachexia (mass index 23 +/- 1 vs. 29 +/- 5 kg/m2, P=0.0005). RESULTS: At baseline, LV end-diastolic volume (197 +/- 78 vs. 203 +/- 65 ml), end-systolic volume (131 +/- 75 vs. 126 +/- 63 ml), LV mass (213 +/- 44 vs. 222 +/- 62 g), and LV ejection fraction (38 +/- 19% vs. 40 +/- 16%) did not differ between cachectic patients and controls (all P>0.10). During follow-up, there was a significant decrease in LV mass in patients with cachexia (-16 g, P<0.05) and a trend to increase in LV mass in patients without cachexia (+7 g, P=0.12, comparison between groups: P=0.010). CONCLUSIONS: The direction of changes over time in LV mass differs in CHF patients with cachexia as compared with non-cachectic controls. A significant decrease in LV mass occurs in patients with cardiac cachexia. This study documents in vivo the occurrence of wasting of the left ventricle in patients with CHF who demonstrate general body wasting. PMID- 15336801 TI - Should oral glucose tolerance test be a routine examination after a myocardial infarction? AB - BACKGROUND: Diabetes mellitus (DM) and impaired glucose tolerance (IGT) are important cardiovascular risk factors. The objective of this study was to assess the prevalence of DM and IGT in patients discharged from the CCU without known DM after a myocardial infarction. METHODS: One hundred and ten patients, men and women aged 31-80 years with a myocardial infarction 1-12 months before inclusion were examined with oral glucose tolerance test. Patients with known DM were excluded. A standard oral glucose test (OGTT) with 75 g of glucose was performed. RESULTS: IGT was observed in 29 (26%) patients and DM in 13 (12%) patients in the OGTT. If only fasting plasma glucose (FPG) was used alone five (38.5%) patients with diabetes subjects and three (10.3%) with IGT were identified. Thus, a FPG test alone identified only 19.0% of the patients with pathological OGTT. The prevalence of DM and IGT in patients discharged from the CCU after a myocardial infarction without known DM diagnosis was high (38%). A fasting glucose alone failed to identify more than 80% of the patients with pathological glucose tolerance in this study. CONCLUSION: Since pathological glucose tolerance is an important cardiovascular risk factor, oral glucose tolerance test should be considered as a routine test after a myocardial infarction in subjects without known DM. PMID- 15336802 TI - Long-term cocaine use is related to cardiac diastolic dysfunction in an African American population in Baltimore, Maryland. AB - BACKGROUND: Only limited studies have been done on the effects of long-term cocaine use on the human heart, and the results remain controversial. In this study, we examined and compared the diastolic function of non-cocaine users and chronic cocaine users to reveal the impact of long-term cocaine use on the human heart. METHODS: Two-dimensional echocardiogram and pulsed Doppler transmitral blood flow pattern were obtained from 138 recruited study participants with different cocaine histories. The indices of cardiac structure and function were measured from the echocardiogram of each participant. Student's t-test was used to compare the average echocardiographic measurements of the non-cocaine user group and the cocaine user group. Multivariate regression analysis was deployed to eliminate the effects of age, gender, blood pressure, and HIV infection on the functional measurements of the two groups. RESULTS: The cocaine user group had a significantly longer average deceleration time than did the non-cocaine user group (208.1 +/- 38.2 vs. 167.5 +/- 39.1 ms, P<0.001). A linear association existed between the deceleration time and the log-transformed duration of cocaine use (beta=0.00351, S.E.=0.00104, P=0.001). Cocaine users in this study were approximately five times more likely to have an elongated deceleration time (>200 ms) than were non-users (OR, 4.799; 95% CI, 1.000-23.044; P=0.05). No significant differences were observed in the other measured diastolic functional parameters, such as isovolumic ventricular relaxation time, E wave, A wave, and E/A ratio. CONCLUSIONS: Long-term cocaine use is linked to decline in diastolic function. PMID- 15336803 TI - Cardiac hypertrophy in hypertension: relation to 24-h blood pressure profile and arterial stiffness. AB - BACKGROUND: In subjects with essential hypertension peripheral blood pressure profile contributes to the pathogenesis of left ventricular hypertrophy. It is not known if central arterial pressure is superior to peripheral blood pressure profile for predicting left ventricular hypertrophy. In the present study 24-h blood pressure profile and central hemodynamics were examined to evaluate mechanical loading factors as determinants of cardiac hypertrophy in mild to moderate untreated essential hypertension. METHODS: Forty-eight untreated subjects with mild to moderate essential hypertension were examined by echocardiography for evaluation of left ventricular mass, 24-h ambulatory blood pressure monitoring (ABPM), and applanation tonometry of the radial artery with pulse wave analysis for evaluation of central hemodynamics. RESULTS: Left ventricular mass showed a statistically significant correlation with age, clinic systolic blood pressure, mean heart rate and heart rate variability during 24-h ABPM, augmentation pressure and index and central systolic blood pressure. In a multiple regression analysis including clinic systolic blood pressure, central systolic pressure, mean systolic pressure and pulse pressure during ambulatory monitoring as well as age, independent predictors of left ventricular mass were only age (P=0.006) and central systolic blood pressure (P=0.04). In conclusion, pulse wave analysis is a valuable method in predicting cardiac hypertrophy in untreated mild to moderate essential hypertension. Central systolic blood pressure should be taken into account for planning therapeutic strategies for prevention of left ventricular hypertrophy in hypertensive patients. PMID- 15336804 TI - Elevated plasma endothelin-1 levels in coronary sinus during rapid right atrial pacing in patients with slow coronary flow. AB - The aim of the study was to evaluate whether there was an imbalance between endothelin-1 (ET-1) and nitric oxide (NOx) release and diffuse atherosclerotic changes existed in patients with slow coronary flow (SCF). Baseline and post atrial pacing coronary sinus ET-1 and NOx levels were measured in 19 patients with SCF (11 female, 56 +/- 9 years) and in 14 control subjects (nine female, 54 +/- 7 years). All patients underwent subsequent intravascular ultrasound (IVUS) investigation at the same setting with right atrial pacing. Baseline arterial (12.4 +/- 9.9 vs. 6.3 +/- 5.1 pg/ml, P<0.005) and coronary sinus (12.2 +/- 11.1 vs. 6.4 +/- 6.9 pg/ml, P<0.005) ET-1 plasma levels were higher in patients than in controls. After atrial pacing, concentration of ET-1 level from coronary sinus (24.7 +/- 14.6) significantly increased as compared to baseline (12.4 +/- 9.9, P<0.0001) and control levels (5.3 +/- 6.3, P<0.0001). Additionally, coronary sinus ET-1 level increased significantly with atrial pacing compared to femoral artery ET-1 level (16.3 +/- 8.5, P<0.005) in patients with SCF. After atrial pacing, the femoral artery ET-1 level also increased in patients compared to control level (P<0.0001). No significant differences in arterial and coronary sinus NOx plasma levels were found between the two groups, both at baseline and after pacing. Upon IVUS investigation, the common finding was longitudinally extended massive calcification throughout the epicardial arteries in patients with SCF. Mean intimal thickness was 0.59 +/- 0.18 mm. The data of this study suggest that increased ET-1 levels and insufficient NOx response, as well as the pathological data of IVUS may be associated with coronary microvascular dysfunction and may be the manifestation of early diffuse epicardial atherosclerosis in these patients with SCF. PMID- 15336805 TI - Hypertrophic cardiomyopathy: electrical abnormalities detected by the extended length ECG and their relation to syncope. AB - BACKGROUND: Ventricular repolarization abnormalities can represent a trigger for lethal arrhythmias in hypertrophic cardiomyopathy (HCM). We sought to assess whether multiparametric computerized surface ECG analysis identifies repolarization abnormalities in HCM patients, and whether this approach allows identification of patients with syncope. METHODS: In 28 HCM patients and 102 healthy subjects (14 and 51 males, mean age 44 +/- 15 and 41 +/- 14 years, respectively), 8-lead ECG (I, II, V1-V6) was recorded for 5 min, acquired in digital format and analyzed. Heart-rate corrected QT (QTc) and T wave complexity index (TWCc), QT dispersion, activation-recovery interval (ARI) and its dispersion, signal duration in the terminal portion of the filtered QRS at 25 Hz (LAS(25 Hz)) were analyzed among other parameters. RESULTS: Compared to healthy subjects, HCM patients exhibited longer QRS, filtered QRS, QTc and QTd, greater TWCc, minor ARId and LA(25 Hz). QRS duration and maximal septum thickness were linearly correlated (r=0.231 p<0.001). ARId shortening depended on ARI shortening in lead V1 (241 +/- 51 vs. 287 +/- 45, HCM vs. healthy subjects, p<0.0001) and lengthening in V6 (257 +/- 42 vs. 209 +/- 34, HCM vs. healthy subjects, p<0.0001). Significant factors for syncope at Wilks' stepwise discriminant analysis were TWCc, QRSd and LAS(25 Hz) (F=14.394, 10.098 and 9.226, respectively) with 92.3% positive predictive accuracy. CONCLUSIONS: In HCM, longer QRS and QT intervals are consequences of increased left ventricular mass, while ARI seems to reflect myocardial activation rather than inhomogeneity of recovery. The simultaneous evaluation of TWC, QRSd and LAS(25 Hz), unable by itself to hold a predictive value, yielded high accuracy in predicting cardiogenic syncope. PMID- 15336806 TI - Telecardiology for patients with chronic heart failure: the 'SHL' experience in Israel. AB - BACKGROUND: Patients with chronic heart failure characteristically have multiple hospital admissions for symptom control, deleteriously affecting their quality of life and imposing a burden on national healthcare costs. We assessed the effect of a novel transtelephonic monitoring and follow-up program on the admission rate and length of hospital stay as well as changes in their subjectively rated quality of life of patients with chronic heart failure. METHODS: This prospective 1-year study was conducted on compliant subscribers to 'SHL', a telecardiological service with >60,000 subscribers, who were admitted > or = 2 times during the previous year for recurrent pulmonary edema or deterioration in heart failure. Their heart rate, blood pressure and body weight measurements were now automatically transmitted daily to 'SHL"s data bank and added to stored and updated medical records. A questionnaire survey acquired information on their quality of life. RESULTS: The study cohort included 118 patients, mean age 75 years (range 49-89 years), 65% males, a II-IV class functional capacity and a 25% (range 10-39%) mean ejection fraction. There was a 66% reduction in the total hospitalization days (from 1623 in the year preceding study entry to 558 during the study period, p<0.0001). Although only 38/118 patients were hospitalized, most participants reported a significant subjective improvement in their quality of life. CONCLUSIONS: Data are provided to demonstrate that a transtelephonic system allowing primary care at the patient's home can significantly reduce hospitalization rate and length of stay and significantly enhance the quality of life of patients with chronic heart failure. PMID- 15336807 TI - Abnormal ventricular repolarization in hypertensive patients: role of sympatho vagal imbalance and left ventricular hypertrophy. AB - BACKGROUND: An increased risk for life-threatening arrhythmias and sudden death has been observed in hypertensive patients, associated with either left ventricular hypertrophy (LVH) or prolonged QT interval. To investigate the influence of autonomic imbalance and LVH on QT interval in hypertensive patients, we compared two different models of LVH: hypertension and endurance physical training. METHODS: Forty-seven untreated subjects affected by essential hypertension and 35 endurance runners, with a similar degree of LVH, were enrolled into the study. All subjects underwent 24-h ambulatory ECG recording and morning blood sampling for catecholamines. Heart rate variability was evaluated by spectral analysis and a computerized algorithm was used to measure the QT interval; QTc was then computed by the Bazett's formula. Left ventricular mass index (LVMI) was assessed by echocardiogram. RESULTS: No difference in LVMI was found between hypertensive patients and athletes. Athletes showed lower heart rate (64 +/- 1 vs. 75 +/- 1 bpm, p<0.001, mean +/- S.E.M.) and shorter QTc (401 +/- 3 vs. 434 +/- 4 ms, p<0.001) than hypertensive patients throughout the 24-h period. Athletes showed a higher vagal drive compared to hypertensive patients as suggested by bradycardia and higher values of vagal indices, which negatively correlated with QTc. Plasma norepinephrine was significantly lower in athletes than in hypertensive patients (p<0.05) and positively correlated with QTc. CONCLUSION: Despite similar degrees of LVH, hypertensive patients show QTc lengthening, as compared to athletes. Heart rate variability and plasma norepinephrine levels suggest sympathetic predominance in hypertensive patients, which could contribute to abnormal ventricular repolarization, thus identifying patients with an increased arrhythmic risk. PMID- 15336808 TI - Effects of age, period and cohort on acute myocardial infarction mortality in Hong Kong. AB - BACKGROUND: Hong Kong has been experiencing a transition from a developing area to a developed area over the past few decades. The time trends of acute myocardial infarction (AMI) mortality would have implications for the future trends of AMI in Mainland China. METHODS: Age-adjusted mortality rates during the period 1976-1999 were calculated by direct standardization with the world population using local mortality and population data. Log-linear regressions were used to estimate the annual percentage changes in mortality over different periods. Poisson regression models were used to explore the effects of age, calendar period and birth cohort. RESULTS: A downward trend was observed for age adjusted AMI mortality rate in both sexes in the 1990s, after the increasing trends in the 1970s and 1980s. The AMI mortality rate among men and women dropped by 36.4% and 37.4%, respectively. A negative annual percentage change was observed across all age groups in both sexes. Both the calendar period and the birth cohort showed significant effects on the changing AMI mortality rate, with the influence of period being stronger than the cohort, especially among males. CONCLUSION: Age-adjusted AMI mortality started to decrease during the last decade. Changing life styles and a reduction of coronary risk factors could have contributed, but the improvements in medical care and effective treatment for patients suffering from AMI should have an important role. PMID- 15336809 TI - Bedside programmed ventricular stimulation for sudden death risk stratification. AB - BACKGROUND: Patients with myocardial infarction and left ventricular dysfunction are at risk for sudden death. This research was conducted to determine the applicability and safety of a bedside programmed stimulation protocol to determine the risk for sudden death in these patients. METHODS: Four hundred and twelve patients with acute myocardial infarction were studied. Left ventricular ejection fraction was evaluated by means of an echocardiogram. Ventricular arrhythmia, late potentials and heart rate variability were determined by means of Holter recordings. Fifty patients (60 +/- 14-year-old; 85% male) presented a left ventricular ejection fraction lower than 0.40 (0.36 +/- 0.10) associated with late potentials, low heart rate variability or ventricular arrhythmia greater than Lown I. After a central venous access was placed under fluoroscopy guidance and ECG monitoring, a quadripolar catheter was advanced to the right ventricular apex to perform programmed ventricular stimulation with up to three extrastimuli. The patients were followed-up to determine in-hospital morbidity and/or mortality. RESULTS: No patient suffered complications. Ventricular tachycardia or ventricular fibrillation was induced in six patients. All of them received amiodarone and in five an automatic cardioverter-defibrillator was implanted. After a 22 +/- 6 month follow-up, five patients had received appropriate discharges from the implanted device and none had suffered from arrhythmic sudden death. CONCLUSION: Bedside programmed stimulation is a safe and useful means for sudden death risk stratification in post myocardial infarction patients. It moreover presents the advantage of being cheaper than conventionally used procedures. PMID- 15336810 TI - Early and late clinical outcomes after primary stenting of the unprotected left main coronary artery stenosis in the setting of acute myocardial infarction. AB - BACKGROUND: Acute left main coronary artery occlusion is a dramatic condition with very high mortality. The study was aimed to evaluate the effect of primary stenting in patients with left main coronary artery (LMCA) disease in the setting of acute myocardial infarction (AMI). METHODS: Between June 1997 and April 2002, primary stenting for left main coronary artery disease was performed in 18 patients with acute myocardial infarction. We evaluated early and late clinical outcomes, and prognostic determinants in this clinical setting. RESULTS: Mean ages of patients were 59 +/- 12 years. Fourteen patients had cardiogenic shock on admission. Angiographic success (TIMI flow > or = 2 and diameter stenosis < 30% after stenting) was achieved in 17 patients (94%). In-hospital death occurred in eight patients (44%). Two patients (11%) received emergent bypass surgery because of hemodynamic instability after primary stenting. On univariate analysis, good pre-intervention TIMI flow (grade > or = 2) was identified as a good prognostic determinant of in-hospital survival. During mean follow-up of 39 +/- 22 months, there was no late death and one patient received bypass surgery. Probability of freedom from death at 3-year was 56 +/- 12%. CONCLUSION: Primary stenting is a valuable therapeutic strategy for left main coronary disease in the setting of acute myocardial infarction, and it might save the life especially in patients with good pre-intervention TIMI flow (grade > or = 2). Long-term clinical outcome of patients surviving to hospital discharge is favorable. PMID- 15336811 TI - Under-diagnosis of diabetes mellitus in patients with coronary artery disease. AB - BACKGROUND: The uptake of medical treatment in patients with coronary artery disease may be suboptimal. Our intention was to review the medical treatment of these patients in the light of current evidence. METHODS: One hundred ninety six consecutive patients with known or suspected coronary artery disease attending a tertiary centre for day case cardiac catheterisation were assessed. Fasting blood samples were sent for glucose and cholesterol. The results of coronary angiography and left ventriculography and any changes in medications were noted. RESULT: One hundred eighty two patients (93%) had fasting blood samples taken. The management of cholesterol in patients with coronary artery disease has improved since 1994. We have demonstrated the benefit of taking fasting blood samples in patients attending for day case angiography: 10% of non-diabetics actually had fasting blood glucose levels of greater than 7.0 mmol/l. All of these newly diagnosed diabetics had coronary artery disease. A further 9% of non diabetics had impaired fasting glycaemia; 69% had coronary artery disease and 8% had left ventricular dysfunction. Among patients with coronary artery disease, there was a statistically significant increase on most categories of medications on discharge compared to admission. For those with left ventricular dysfunction, there was a statistically significant increase in the use of Angiotensin Converting-Enzyme (ACE)-inhibitors. CONCLUSION: Diabetes mellitus is under diagnosed in patients with coronary heart disease. They are at higher risk of coronary artery disease and therefore need intensive management. Testing all patients attending for day case cardiac catheterisation for fasting glucose would enable diagnosis and initiation of treatment of a high-risk group. PMID- 15336812 TI - Wide QRS complex tachycardia. Rapid method of prognostic evaluation. AB - A wide QRS complex tachycardia suggests a ventricular tachycardia (VT); but supraventricular tachycardia (SVT) is also possible. Some authors reported on the electrocardiographic signs for the differential diagnosis of VT and SVT with aberrancy. Frequently these signs are debatable and the diagnosis is uncertain. The purpose of the study was to evaluate the interest of a non-invasive study by transesophageal route for the evaluation of the nature of a wide QRS complex tachycardia in which a reliable ECG algorithm does not permit to distinguish VT from SVT with aberrancy. METHODS: Esophageal electrophysiologic study (EPS) was performed in 53 patients, aged from 16 to 85 years without bundle branch block (BBB) in sinus rhythm, but with wide-QRS tachycardia. The protocol consisted of atrial pacing at progressively higher rates and then programmed stimulation with one and two extrastimuli in control state and after isoproterenol infusion. Intracardiac EPS was performed in 49 of them. RESULTS: (1) Study was negative in nine patients; intracardiac EPS remained negative in four of them, induced a VT in five; (2) clinical tachycardia was induced in 44 patients: (a) in 29 of them, atrial pacing induced a BBB similar to aberrancy noted in tachycardia and the diagnosis of SVT with aberrancy was made; (b) in 15 patients, QRS complex remained narrow during atrial pacing; the diagnosis of VT was made in presence of AV dissociation and confirmed by intracardiac study. VT was induced by atrial or ventricular stimulation or was spontaneous during isoproterenol infusion. VT mechanism were bundle branch reentry [Am. J. Cardiol. 65 (1990) 322], verapamilsensitive VT [Am. J. Cardiol. 65 (1990) 322], catecholamine-sensitive VT [J. Cardiovasc. Electrophysiol. 7 (1996) 2]. Two patients had tachycardias of both natures either supraventricular or ventricular. CONCLUSION: Esophageal EPS was a safe, rapid and economic means to evaluate the mechanism of wide QRS tachycardia in 84% of patients; atrial pacing at progressively higher rates is very simple to reproduce the aberrancy of similar morphology in those patients who had wide-QRS tachycardia related to a SVT with aberrancy. If atrial pacing did not exactly reproduce the aberrancy in tachycardia, a VT should be suspected. PMID- 15336813 TI - Left ventricular hypertrabeculation/noncompaction with and without neuromuscular disorders. AB - BACKGROUND: Left ventricular hypertrabeculation/noncompaction (LVHT) is frequently associated with neuromuscular disorders (NMD). It is unknown, whether LVHT patients differ according to the presence or absence of NMD. Aim of the study was to assess, if clinical, ECG or echocardiographic findings differ between LVHT patients with and without NMD. METHODS: Included were all patients, in whom LVHT was diagnosed between June 1995 and February 2003 in one echocardiographic laboratory. All patients underwent a cardiologic examination and were invited for a neurologic investigation. RESULTS: Of 77 patients with LVHT (19 female, mean age 52 years), 59 were investigated neurologically. Eleven were neurologically normal, 21 had a definite NMD (metabolic myopathy, n=15; Leber's hereditary optic neuropathy, n=3; myotonic dystrophy, n=2 and Becker muscular dystrophy, n=1). The remaining 27 had a NMD of unknown etiology. Neurologically normal patients had more often anginal chest pain than patients with definite NMD (64% vs. 14%, P=0.0042) or NMD of unknown etiology (64% vs. 26%, P=0.0157). Neurologically normal patients were more often in NYHA class 0 or I than patients with NMD of unknown etiology (64% vs. 26%, P=0.0289) and had a thinner interventricular septum than patients with NMD (10.6 mm vs. 12.8 mm, P=0.0253). CONCLUSIONS: Cardiac abnormalities are hardly different between patients with and without NMD. PMID- 15336814 TI - Efficiency of L-arginine enriched cardioplegia and non-cardioplegic reperfusion in ischemic hearts. AB - OBJECTIVE: Endothelial dysfunction with decreased nitric oxide (NO) levels has been implicated on reperfusion injury. Although L-arginine has been shown to diminish reperfusion injury in in vitro studies, clinical studies were very limited. METHODS: Forty patients with acute myocardial ischemia undergoing CABG were randomized to a study and a control group. L-Arginine was added to cardioplegia solutions in study group. A non-cardioplegic warm blood solution with 8 mmol/l L-arginine infused for controlled reperfusion. Control patients received same protocol without L-arginine. Myocardial O2, lactate, nitrite and malondialdehyde extractions were measured in addition to calculation of CK-MB/CPK ratio and hemodynamic data. RESULTS: While there was no mortality in study group, one patient in control group died. Overall and nitrite (P=0.01) and lactate extractions (P=0.04) was higher in study and control groups, respectively. Myocardial O2 uptake was higher and malondialdehyde extraction was lower in study group. CK-MB/CPK ratio at postoperative sixth hour was also significantly lower in study group. Ninety percent of the study group had spontaneous return of the sinus rhythm, while 80% of the control patients required defibrillation (P<0.0001). In addition to significantly better hemodynamics, perioperative myocardial infarction incidence was lower (P=0.037), the length of intensive care unit (P=0.009) and hospital (0.014) stays were shorter in study group. CONCLUSIONS: Use of L-arginine for protection of acutely ischemic myocardium appears to be a safe technique. L-Arginine supplementation increased NO levels and attenuated free O2 radical mediated myocardial injury. Controlled reperfusion with l-arginine enriched non-cardioplegic blood could be a new therapeutic entity to diminish ischemia/reperfusion injury. PMID- 15336815 TI - C-reactive protein concentrations are related to insulin resistance and metabolic syndrome as defined by the ATP III report. AB - BACKGROUND: C-reactive protein (CRP), very sensitive acute phase reactant, is an important marker of coronary artery disease. However, the relationship between insulin resistance and CRP has not been thoroughly studied. We observed the association between CRP, insulin resistance and metabolic syndrome as defined by the ATP III report, and thus identified the role of CRP in the relation to insulin resistance. METHODS: Seven hundred and sixty-seven subjects (436 men, 331 women) who underwent a medical check-up at health promotion center in a University Hospital during March 2002, aged 20-84 years, were included in this study. The components of metabolic syndrome as defined by the ATP III report and high sensitivity CRP levels were analyzed, and Homeostasis model assessment index (HOMA) and quantitative insulin sensitivity check index (QUICKI) were calculated. RESULTS: The mean concentrations of CRP in subjects according to the presence of 0, 1, 2, 3, 4, or 5 components of metabolic syndrome as defined by ATP III were 0.64, 0.95, 1.14, 1.19, 2.40, and 2.53 mg/l, respectively. The mean concentrations of CRP were significantly higher in subjects with a high insulin resistance (higher HOMA index and lower QUICKI) than in those with a low insulin resistance. Significant positive correlations were identified between CRP and BMI, waist circumference, triglyceride, blood pressure, glucose and HOMA index. A significant negative correlation was found between CRP and HDL cholesterol or QUICKI. CONCLUSION: These results suggest that metabolic syndrome and insulin resistance are associated with systemic inflammatory response, which plays an important pathogenic role in atherosclerosis. PMID- 15336817 TI - Effects of a weight reduction program with and without aerobic exercise in the metabolic syndrome. AB - BACKGROUND: In the setting of a rural, primary care practice, evidence for the efficacy of intentional weight loss with or without aerobic exercise training (AET) in hypertensive patients with the metabolic syndrome remains controversial. DESIGN OF STUDY, METHODS: We analysed data of 52 hypertensives with the metabolic syndrome, who attended a weight management program with or without AET (diet/exercise group, n = 18; diet group, n = 20) for 36 months. Patients with a similar risk profile, who declined antihypertensive therapy beyond ACE inhibition, served as controls (n = 14). RESULTS: Body mass index (BMI) was significantly reduced over time in the diet and diet/exercise group (DeltaBMI: 7.2 +/- 1.1 and -6.6 +/- 1.5 kg/m2) vs. the control group (DeltaBMI: +0.4 +/- 1.3 kg/m2; p < 0.001 at 36 m). While systolic and diastolic blood pressures (BPs) did not change over time in controls (175 +/- 5/89 +/- 7 mm Hg), BPs were significantly reduced in the diet group (151 +/- 8/75 +/- 10 mm Hg) and in the diet/exercise group (139 +/- 12/71 +/- 8 mm Hg; p < 0.001 vs. diet group). Metabolic abnormalities were significantly improved over time in the diet group vs. controls, while AET did not add further benefits. However, heart rate recovery after acute exercise was improved and body cell mass (BCM) was increased in the diet/exercise group vs. patients without AET (p < 0.001 vs. others). CONCLUSIONS: Weight management significantly improves the lipid and nonlipid abnormalities of the metabolic syndrome, which is associated with reduced blood pressure. Addition of AET does not add further benefits on metabolic parameters, but improves blood pressure regulation. Effective lifestyle modifications are feasible even in the setting of a rural practice. PMID- 15336816 TI - Randomized, controlled trial of secondary prevention of coronary sclerosis in normocholesterolemic patients using pravastatin: final 5-year angiographic follow up of the Prevention of Coronary Sclerosis (PCS) study. AB - BACKGROUND: The potential benefit of cholesterol-lowering therapy for normocholesterolemic patients with coronary artery disease (CAD) has not been clarified. The Prevention of Coronary Sclerosis (PCS) Study was designed to evaluate the effect of pravastatin on secondary prevention of progression of CAD in normocholesterolemic patients for a period of 5 years. METHODS: A total of 329 patients with CAD were enrolled. Normocholesterolemic patients were defined by a serum total cholesterol (TC) level of 180-219 mg/dl. Patients in this group were randomized into pravastatin and dietary control groups. Patients whose serum TC level fell outside the normal range were divided into a high-cholesterol reference group (TC > or = 220 mg/dl) and a low-cholesterol reference group (TC < 180 mg/dl). Patients in the pravastatin and high-cholesterol groups received pravastatin 10 mg/day. Coronary angiography was performed at baseline, 2 years, and 5 years and analyzed by quantitative coronary arteriography. Angiographic coronary progression was evaluated by minimum obstruction diameter (MOD) and mean segment diameter (MSD). RESULTS: At 5 years, change in MOD was significantly (change, P=0.033; change/years, P=0.032) less in the pravastatin group (-0.04 +/- 0.17 mm) than in the dietary control group (-0.16 +/- 0.27 mm). Although a similar trend was observed in the MSD results, the differences were not significant. CONCLUSION: Long-term angiographic data show that cholesterol lowering therapy by pravastatin prevents progression of coronary atherosclerosis in normocholesterolemic patients with CAD. PMID- 15336818 TI - Lack of regeneration of myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans with large anterior myocardial infarctions. AB - BACKGROUND: Experimental and preliminary clinical data suggest that transplantation of autologous bone marrow cells (BMC) may contribute to regeneration of the myocardium after acute myocardial infarction. This approach should be tested in patients with large infarctions in whom a positive effect would be most beneficial. METHODS AND RESULTS: After successful recanalization within 5.9 +/- 2.5 h and stent implantation in five patients with a large acute anterior myocardial infarction (AMI), the patients received autologous mononuclear BMCs via a balloon catheter placed into the left anterior descending artery 6.3 +/- 0.4 days after revascularization. At 3-month follow-up, no improvement was observed for left ventricular ejection fraction, regional wall motion in the infarcted zone, contractility index measured via dobutamine stress echocardiography, coronary blood flow reserve and maximal oxygen uptake, respectively. After further follow-up of 12 months, again no change of the left ventricular ejection fraction could be detected. CONCLUSIONS: Intracoronary transplantation of autologous mononuclear BMCs did not improve cardiac function in our patients with large anterior myocardial infarctions after 3 and 12 months. PMID- 15336819 TI - New insight into "heart-hand" syndromes: a newly discovered chromosomal abnormality in a family with "heart-hand" syndrome. AB - Heart-hand syndromes are a genetic heterogeneous family of disorders, which are characterised by congenital cardiac and forelimb anomalies. We describe a deletion on chromosome 6, which seems to be connected to the phenotypic expression of a heart-hand syndrome in many affected members of a family who, in addition to the characteristic hypoplastic, manifested hyperplastic skeletal abnormalities. This newly discovered chromosomal abnormality confirms once more the genetic heterogeneity of the syndrome. PMID- 15336820 TI - Recurrent unstable angina after revascularization in a case of dual left anterior descending coronary artery without risk factors: casualness or destiny? PMID- 15336821 TI - Transcatheter embolization of multiple bilateral pulmonary arteriovenous malformations in hereditary hemorrhagic telangiectasia. PMID- 15336822 TI - Pathogenesis of postinfarction papillary muscle rupture. PMID- 15336823 TI - Isolated true contractile left ventricular diverticulum in an adult patient. PMID- 15336824 TI - Ventricular fibrillation during sleep in an adolescent with hypertrophic cardiomyopathy: the difficulty of risk stratification and the power of the cardioverter-defibrillator. PMID- 15336825 TI - Coronary perforation during unprotected left main angioplasty. Management with conservative approach: a case report. PMID- 15336826 TI - Arrhythmogenic biventricular dysplasia/cardiomyopathy masquerading as dilated cardiomyopathy with typical electrocardiographic features. PMID- 15336827 TI - Myocardial infarction and HELLP: a case of heightened vasomotor reactivity. PMID- 15336828 TI - PCNA expression in ischemic cardiomyopathy: DNA repair, myocyte regeneration or just another type of myocyte death? PMID- 15336829 TI - Neutrophil count and infarct size in patients with acute myocardial infarction. PMID- 15336830 TI - Pseudoaneurysm complicating aortic coarctation in a pregnant woman. PMID- 15336831 TI - Successful treatment of four-valve native endocarditis caused by Streptococcus bovis. PMID- 15336832 TI - Emergency pacemaker implantation in acromegaly. PMID- 15336833 TI - Mixed apical left ventricular hypertrophic cardiomyopathy with surrenal adenoma. PMID- 15336834 TI - From myth to reality: lessons learned from the first experiments. PMID- 15336835 TI - Cardiac restoration by cell transplantation. PMID- 15336836 TI - Cell-based myocardial repair: how should we proceed? AB - Cell-based myocardial repair and regeneration heralds a new frontier in the treatment of cardiovascular disease. It provides an unprecedented opportunity to treat the underlying loss of cardiomyocytes that occurs after myocardial injury and that results in the cascade of events leading to heart failure. Yet, even as it progresses to the clinic, much remains to be understood about this technology. For example, controversies exist over the specific cells to be used, the cell dosages needed, how cells will impact the electrical activity of the myocardium, and even whether transplanted cells can actually improve myocardial function. We can perhaps answer these questions more quickly and more effectively - and thus benefit patients more rapidly - if we learn from the successes and failures of our gene therapy colleagues and take a prudent, step-wise approach from bench to bedside. To do so, we need only to promise what we can deliver, to do careful science, and then to deliver well on our promises. Although cellular cardiomyoplasty (cell transplantation for cardiac repair) shows great early clinical promise, its future as a new frontier in the treatment for cardiovascular disease will rest heavily on how we move forward in the next few years. Its success will heavily depend upon conducting carefully controlled, randomized double-blind clinical trials with appropriate endpoints, in the right patients. Choice of cell type, and mode of cell delivery, will also have to be considered, and may have to be matched to the patient. Irrespective of cell type, we can also be assured that cells offer both an opportunity for tissue repair and the potential for not yet understood outcomes. As with any frontier, there will be pitfalls and consequences to be considered that may surpass those of previous endeavors. But so too is the potential for previously unimagined success at treating the leading cause of death in the western world. In short, the promise for cardiovascular cell therapy is too great to be spoiled by ill-designed attempts that forget to account for both the natural propensities of cells and of the myocardium. PMID- 15336837 TI - Factors related to the efficacy of skeletal muscle cell transplantation and future approaches with control-released cell growth factors and minimally invasive surgery. PMID- 15336838 TI - The strength of plasticity: stem cells for cardiac repair. PMID- 15336839 TI - Human stem cells shape the future of cardiac regeneration research. PMID- 15336840 TI - Role of stem cell homing in myocardial regeneration. AB - Recent studies have demonstrated the potential for myocardial regeneration at the time of myocardial infarction. Our lab focuses on understanding how stem cells home to injured tissue. We believe that a detailed analysis of the biological responses to tissue injury not only gives us insight into how damage occurs, but can also yield insight into how the body attempts to repair itself. In the case of the heart, we have shown that these "healing pathways" are only expressed for a short time following injury, and this may be why myocardial damage is usually irreversible. Furthermore, work suggests that deciphering these "healing pathways" and re-establishing their expression at time remote from injury offers an avenue for developing novel therapeutics that ultimately may allow us to orchestrate and exaggerate the healing process at time remote from tissue injury. PMID- 15336841 TI - The regenerative potential of the human heart. PMID- 15336842 TI - Autologous human serum for cell culture avoids the implantation of cardioverter defibrillators in cellular cardiomyoplasty. AB - BACKGROUND: Current clinical experience with cellular cardiomyoplasty (using serum bovine-cultivated myoblasts) has demonstrated significant malignant ventricular arrhythmias and sudden deaths in patients. In some ongoing clinical trials the implantation of cardioverter-defibrillator is mandatory. We have hypothesized that contact of human cells with fetal bovine serum results after 3 week fixation of animal proteins on the cell surface, representing an antigenic substrate for immunological and inflammatory adverse events. METHODS AND RESULTS: Autologous myoblasts were transplanted into infarcted LV in 20 patients (90% males, mean age 62+/-8 years). Cells were cultivated in a complete human medium during 3 weeks, using the patients' own serum obtained from a blood sample or from plasmapheresis. Injections were performed during CABG (2.1 grafts/pt). All patients had an uneventful recovery. At a mean follow-up of 14 +/- 5 months without mortality, no malignant cardiac arrhythmias are reported. LV ejection fraction improved from 28 +/- 3% to 52 +/- 4.7% (p = 0.03), and regional wall motion score index (WMSI) from 3.1 to 1.4 (p = 0.04) in the cell-treated segments. Myocardial viability tests showed areas of regeneration. Patients moved from mean NYHA class 2.5 to class 1.2. CONCLUSIONS: A total autologous cell culture procedure was used in cellular cardiomyoplasty reducing the risk of arrhythmia. Human-autologous-serum cell expansion avoids the risk of prion, viral or zoonoses contamination. Since patients treated with noncultivated bone marrow cells are free of arrhythmia, the bovine-culture medium seems to be responsible for this complication. Cellular cardiomyoplasty may be efficient to avoid progression of ventricular remodeling and subsequent heart failure in ischemic heart disease. PMID- 15336843 TI - Endothelial progenitor cells: a potential versatile tool for the treatment of ischemic cardiomyopathies -- a clinician's point of view. PMID- 15336844 TI - Cell therapy: an evolutionary development of transfusion medicine. PMID- 15336845 TI - Xenogeneic cell transplant: fact or fancy? PMID- 15336846 TI - Transendocardial injection of autologous mononuclear bone marrow cells in end stage ischemic heart failure patients: one-year follow-up. PMID- 15336847 TI - Transvascular cellular cardiomyoplasty. PMID- 15336848 TI - Echocardiographic assessment in cell transplantation. PMID- 15336849 TI - Nuclear imaging after cell implantation. AB - Since no currently available method is able to determine engraftment into the myocardium or stem cell differentiation in a living patient, the evidence of stem cell therapy efficacy is in all cases indirect. PMID- 15336850 TI - A new approach to heart valve tissue engineering. PMID- 15336851 TI - Heart valve tissue engineering -- where do we stand? PMID- 15336852 TI - Cardiac tissue repair - cell transplant and growth factors. Proceedings of a meeting. June 4-5, 2004. Sirmione, Italy. PMID- 15336855 TI - Seven challenges for neurosurgeons and physicians in the next decade. PMID- 15336856 TI - Cytological and bacteriological studies of intraoperative autologous blood in neurosurgery. AB - BACKGROUND: To ensure the safety of salvaged blood in neurologic surgery, reinfused blood through the Cell Saver System (CSS) (Hemonetics) was investigated cytologically and bacteriologically. METHODS: Specimens of reinfused blood were cytologically examined with Papanicolaou or Giemsa stains. Reinfused blood and air in the operating theater were investigated by microbiologic techniques. The concentration of dust particles in the theater was determined. RESULTS: Tumor cells were positive in reinfused blood in 5 of 9 specimens with glioblastoma, in 2 of 8 with pituitary adenoma, and 1 of 13 with meningioma. The probability of migration of meningioma cells into reinfused blood was significantly low in comparison with that of glioma cells. Of the 30 specimens studied microbiologically, the bacterial growth was detected in salvaged blood of 14 specimens (46.7%) and in the air of the operating theater for 8 specimens (26.7%). In craniotomy, the contamination rate was 10 of 26 specimens of reinfused blood (38.5%). Most microorganisms were found to be staphylococci. No statistically significant correlation could be found between salvaged blood and air as to contamination or between reinfused blood and the concentration of dust particles in the theater as to bacteriologic results. No infectious complications were found after the operation, though salvaged blood through the CSS was reinfused in 37 patients without glioblastoma or transsphenoidal approach. CONCLUSIONS: The CSS cannot always entrap tumor cells. Salvaged blood should not be reinfused in the patients with glioblastoma or transsphenoidal surgery. None of the patients with reinfusion had any infectious complications. Reinfusion of salvaged blood seems to be safe in neurosurgery. PMID- 15336858 TI - Posttraumatic basilar artery vasospasm. AB - BACKGROUND: During the past decade, vasospasm following posttraumatic subarachnoid hemorrhage (tSAH) has drawn increasing attention. However, despite accumulating evidence linking this phenomenon with poorer outcome, the clinical significance of posttraumatic vasospasm is still debated and often disputed, so that no definite therapeutic attitude has yet been adopted. Recent attention has been drawn to basilar vasospasm, suggesting a negative influence on neurologic outcome. The aim of the present study was the evaluation of basilar vasospasm as an independent factor of secondary brain damage following posttraumatic hemorrhage. METHODS: Daily transcranial Doppler (TCD) evaluations were conducted in 93 consecutive patients with tSAH. Basilar artery (BA) vasospasm was defined by blood flow velocity (FV) higher than 85 cm/s for at least 2 consecutive days. RESULTS: Thirty-two patients (34.4%) had BA FVs higher than 85 cm/s. In those patients, the ratio between BA FV and mean vertebral artery FV was higher than 2.5 and below 2 and those without TCD signs of vasospasm. Using multivariate logistic regression analysis, BA vasospasm proved to independently influence neurologic outcome. CONCLUSIONS: BA vasospasm severe enough to compromise cerebral blood flow to the brainstem, although uncommon, may nevertheless have a potential deleterious effect on neural tissue sensitized by trauma. The present results suggest that specific imaging procedures and eventually therapeutic measures should be conducted in the presence of significant BA vasospasm. PMID- 15336860 TI - Survival trends in elderly patients with glioblastoma multiforme: resective surgery, radiation, and chemotherapy. AB - BACKGROUND: It is appropriate to investigate and to determine survival trends following glioblastoma multiforme treatment using resective surgery, radiation therapy, and/or chemotherapy in patients aged 59 years and higher. METHODS: We retrospectively reviewed 30 elderly patients (> or =59 years old) who were treated for histopathologically confirmed glioblastoma multiforme at our tertiary care institution from 1990 through 2002. All patients were treated with steroids. In addition, 22 patients underwent resective surgery (RS), 17 patients underwent radiation therapy (RT), and 10 patients underwent chemotherapy (C). Many patients underwent these treatments in various combinations: 6 underwent biopsy only, 7 RS only, 6 RS+RT only, and 9 RS+RT+C. For each case, pre-treatment Karnofsky performance scores (KPS), tumor location, presenting symptoms and signs, associated surgical morbidity, and pre-existing medical conditions were also recorded. Patients were categorized into one of four treatment subgroups: Biopsy only, RS only, RS+RT, and RT+RS+C. For each of these subgroups, pretreatment KPS and post-treatment survival were compared. RESULTS: Post-treatment survival following biopsy only was 3.2 +/- 0.8 months (mean +/- SE); RS 2.2 +/- 0.5; RS+RT 5.5 +/- 1.2; RS+RT+C 13.6 +/- 2.1. A longer survival trend was noted for the RS+RT versus RS group (two-tailed unpaired t test, p = 0.02;), as well as the RS+RT+C group, which showed consistently higher survival in comparison to most of the other groups (p = 0.0021, 0.00039, 0.013 vs. the biopsy only, RS only, and RS+RT groups, respectively). No significant difference was found in KPS, comparing all individual groups versus each other (p > or = 0.06). Remarkably, 6 patients survived over 14 months (range, 14.1-22.7 months), all of which received RS+RT+C. CONCLUSIONS: This study suggests a significant improvement in elderly patients treated with the combination of resective surgery, radiation therapy, and chemotherapy, rather than either treatment alone or other combination. This significant improvement does not appear to be biased by pretreatment KPS, as mean KPS values did not significantly differ between any of these groups. However, a greater number of patients in each group must be considered to achieve the power to make more definitive treatment guidelines. PMID- 15336862 TI - Location of the active contact within the subthalamic nucleus (STN) in the treatment of idiopathic Parkinson's disease. AB - BACKGROUND: Chronic electrical stimulation of the subthalamic nucleus (STN) has been shown to be safe and effective in the treatment of medically refractory idiopathic Parkinson's disease. The clinically most relevant location of stimulation within the physiologically defined STN has not been confirmed. We reviewed the locations of active electrical contacts in 33 patients who underwent simultaneous bilateral STN deep brain stimulator (DBS) implantation. METHODS: The location of the microelectrode-defined dorsal STN border was compared to the location of the center of the active contact(s) employed in achieving optimal clinical results 6 to 18 months postoperatively. Furthermore, the location of this optimal contact was determined with respect to each individual patient's midcommissural point. Bilateral monopolar stimulation was employed in 30 patients using quadripolar DBS electrodes. RESULTS: After a minimum follow-up period of 6 months, the motor subscores (UPDRS Part III) in the postoperative on stimulation/off-medication state were 64 +/- 18% (mean +/- SD) improved as compared to the preoperative off-medication state (p < 0.01). Additionally, an improvement of 53 +/- 38% was noted when comparing the postoperative on stimulation/on-medication state to the preoperative on-medication state (p < 0.01). On average, the center of the optimal contact was 13.3 mm lateral, 0.5 mm posterior, 0.1 mm inferior to the mid-commissural point and was 0.1 +/- 2.1 mm dorsal to the physiologically defined dorsal STN border. CONCLUSIONS: While the achieved clinical results are comparable to those published in the literature, it appears that monopolar electrical stimulation at the anterior dorsolateral border of the STN yields optimal clinical results. Further studies are crucial in determining the precise mechanism of various modes of DBS in an effort to maximize clinical outcome. PMID- 15336865 TI - Sellar repair with fibrin sealant and collagen fleece after endoscopic endonasal transsphenoidal surgery. AB - OBJECTIVE: To determine, in patients undergoing sellar repair after endoscopic endonasal transsphenoidal surgery, the clinical efficacy of a combination of fibrin sealant/collagen fleece compared to the use of fibrin sealant or collagen fleece alone, in preventing CSF-related (cerebrospinal fluid) postoperative complications. METHODS: From a retrospective analysis of our series of 242 consecutive endoscopic transsphenoidal procedures, in 56 out of the 90 cases in which the sella had been repaired, fibrin sealant and/or collagen fleece was employed, both in combination with one or multiple layers of other materials. The incidence of postoperative CSF leaks and the need for a postoperative lumbar drainage in the groups of fibrin sealant or collagen fleece treated patients were compared to the group of patients treated with the fibrin sealant/collagen fleece combination. RESULTS: In 2 out of 16 fibrin sealant treated patients a postoperative CSF leak presented, and in 6 out of these 16 subjects a postoperative lumbar drainage was necessary; patients who received a fibrin sealant/collagen fleece combination exhibited no detectable postoperative CSF leak, and no postoperative lumbar drainage was used. CONCLUSIONS: Closure of the sella turcica with fibrin sealant in combination with a collagen fleece is a safe and effective method to prevent CSF fistulas. When used in combination, the collagen fleece enhances the sealing and tissue regeneration properties of the fibrin sealant, thus reducing the incidence of postoperative CSF leaks, obviating the need for a lumbar drain placement. PMID- 15336867 TI - Diagnosis and treatment of Candida vertebral osteomyelitis: clinical experience with a short course therapy of amphotericin B lipid complex. AB - BACKGROUND: Musculoskeletal candidiasis occurs in some patients with candidemia resulting from organ infection, IV drug use, or indwelling central venous catheters. Diagnosis is often difficult because of vague symptomatology and a frequent afebrile course. CASE DESCRIPTION: Three patients with Candida vertebral osteomyelitis are presented. All followed the use of indwelling central venous access catheters and antimicrobial therapy between 6 months and 3 years earlier. In 2, fungemia with the same Candida spp. preceded the spondylodiskitis. These 3 patients bring to nearly 75 the number of reported individuals with what was once quite rare. Although IV amphotericin B doxycholate and fluconazole have usually been effective therapy over prolonged periods of time, we used liposomal amphotericin B to treat 2 of our 3 patients. Both received 5 mg/kg daily for 18 42 days that resulted in total disappearance of signs and symptoms. CONCLUSION: This relatively brief duration of therapy reduces treatment time and is cost effective. PMID- 15336869 TI - Attenuation of vascularity by preoperative radiosurgery facilitates total removal of a hypervascular hemangioblastoma at the cerebello-pontine angle: case report. AB - BACKGROUND: The surgical removal of solid, large, and deep-seated hemangioblastomas remains challenging because it is difficult to control bleeding during the procedure. We used preoperative radiosurgery in a solid, highly vascular hemangioblastoma at the left cerebello-pontine angle and present our angiographic, operative, and histologic findings. CASE DESCRIPTION: A 37-year-old paraplegic woman with multiple hemangioblastomas was re-admitted to our clinic with cerebellar ataxia 6 years after resection of a tumor at the fourth ventricle. A vertebral artery angiogram revealed that the 3.5 cm diameter hemangioblastoma at the left cerebello-pontine angle was highly vascular and fed by the left anterior inferior cerebellar artery and posterior inferior cerebellar artery. Nine months before surgical removal it was treated with stereotactic radiosurgery (gamma knife, margin dose 28 Gy) to inhibit tumor progression and to reduce its vascularity. The tumor was totally removed via the left lateral suboccipital approach; bleeding was well controlled and there were no complications. Pathologic examination of the content of the excised tumor revealed coagulation necrosis with hyaline degeneration of the tumor vessels, resulting in a marked decrease in its vascularity. CONCLUSION: Preoperative radiosurgery led to a marked reduction in the vascularity of this hypervascular hemangioblastoma and was useful for controlling bleeding from the tumor during resection. We succeeded to remove the vascular-rich hemangioblastma after the intentional preoperative radiosurgery. The pathologic changes induced by radiotherapy were confirmed by operative finding. PMID- 15336872 TI - Facial nerve schwannomas: different manifestations and outcomes. AB - BACKGROUND: The purpose of this study was to provide data on the different clinical presentations of facial nerve schwannoma, the appropriate planning for the management of schwannoma of various origins, and the predictive outcomes of surgical management. METHODS: A retrospective study was conducted in a tertiary referral hospital. We reviewed 8 consecutive cases of facial nerve schwannoma diagnosed and managed between 1993 and 2001. RESULTS: Facial nerve schwannomas originated in the internal auditory canal (IAC) (2 cases), parotid gland (2 cases), intratemporal portion (3 cases), and stylomastoid foramen (1 case). Tumor of the stylomastoid foramen presented as an intra- and extratemporal mass. The initial presenting symptom of the 8 patients was facial nerve paralysis in 4 patients, hearing loss in 2, facial numbness in 1, and an infra-auricular mass in 1. Facial palsy occurred in 7 patients during the course of the disease. One patient with a mass in the parotid gland did not show facial palsy up to 1 year after presentation of the initial symptom (facial numbness). Facial nerve paralysis was most severe in intratemporal tumors and less severe in parotid tumors. The patients with IAC suffered from hearing loss and intermittent vertigo and showed decreased vestibular function. The patients with intratemporal tumors also complained of hearing loss. The tumors were completely removed by superficial parotidectomy for parotid tumors; the translabyrinthine approach for 1 IAC tumor and 1 intratemporal tumor; the middle fossa approach for the other IAC tumor; the transmastoid approach for mastoid tumors; and the infratemporal fossa approach for intratemporal and extratemporal tumors. End-to-end cable grafts for the facial nerve were performed in 5 out of 8 cases. In 2 cases, the facial nerve was preserved after the resection of the mass. One case showed complete loss of the peripheral branch of the facial nerve. CONCLUSIONS: Facial nerve schwannoma can present in various ways. By examining the site of origin and the presenting symptoms and signs, we were able to diagnose facial nerve schwannoma preoperatively. According to the operative management of the facial nerve, the postoperative outcome of facial function could be estimated. Our finding could be pivotal in the management of the facial nerve schwannoma. PMID- 15336874 TI - Arteriovenous malformation of the scalp: case report and review of the literature. AB - BACKGROUND: Arteriovenous malformations (AVMs) of the scalp are relatively rare, and their precise natural course remains to be elucidated. We encountered a patient with a scalp AVM that progressively enlarged over the course of 3 years by capturing feeders from intracranial arteries. To our knowledge, ours is the first serial angiographic depiction of the growth of a scalp AVM and the development of a de novo aneurysm in the superior temporal artery (STA) that performed as a feeder. CASE DESCRIPTION: This 21-year-old female consulted us in 1998 complaining of right tinnitus and a pulsating mass in the retroauricular region. The initial angiogram revealed an AVM in the right temporo-parietal subcutaneous space with feeders from the STA, an occipital artery, a posterior auricular artery, and a middle meningeal artery (MMA). Three years later, she complained of enlargement of the lesion, increased tinnitus, and alopecia. Repeat angiographic study revealed the presence of a nidus and the appearance of new feeders from a contralateral MMA and an ipsilateral middle cerebral artery; there was a de novo saccular aneurysm in the right STA. On the day preceding surgery, the left MMA was embolized to control intraoperative bleeding. The AVM was removed totally without any dermal complications. CONCLUSION: This case suggests that scalp AVMs can become enlarged by capturing subcutaneous or intracranial feeders, and that the consequent hemodynamic stress may induce de novo aneurysms in scalp AVMs. Capillary endothelial cells were strongly immunostained for vascular endothelial growth factor. PMID- 15336875 TI - Fatal bihemispheric intracerebral hemorrhage after burrhole evacuation of chronic subdural hematoma in a bone marrow-transplanted patient: case report and review of the literature. AB - BACKGROUND: Intracranial hemorrhage seems to be a relatively frequent complication following bone marrow transplantation (BMT). The neurosurgical management of these patients can be problematic. CASE DESCRIPTION: We report about a 46-year-old patient, who had BMT because of a Pre-T-ALL and who developed symptomatic bihemispheric chronic subdural hematomas (cSDHs). Pre-T-ALL is a form of an acute lymphatic leukemia of T cell type characterized by certain immunologic markers with a worse prognosis. A few hours after evacuation of the cSDHs, the patient developed a fulminant intracerebral hemorrhage on each hemisphere and expired 2 days later. We reviewed the literature reporting similar cases to discuss the indication for a neurosurgical intervention. CONCLUSION: We presume a higher incidence of intracranial hemorrhage in BMT patients than reported. This is caused by the pathologically altered parenchyma related to the extensive therapy in these patients. BMT patients are typically at high risk for intracranial bleeding and are treated conservatively or surgically. We cannot generally recommend the conservative management for symptomatic subdural hematomas/hygromas. The risk for an acute worsening, secondary to repeated hemorrhage, is equivalent to the risk of surgical intervention. Although the outcome of our case is tragic, it should not deter surgical intervention. PMID- 15336877 TI - Inflammatory pseudotumor of the pituitary: case report. AB - BACKGROUND: Inflammatory pseudotumor of the pituitary is a very rare nonneoplastic lesion. We describe a case of a patient with past history of lymphocytic meningitis. CASE DESCRIPTION: A 32-year-old man presented with polyuria, polydipsia, anorexia, abdominal discomfort, and panhypopituitarism. He had 2 episodes of lymphocytic meningitis in the last two years. Magnetic resonance image (MRI) disclosed a sellar and suprasellar mass with extension to the pituitary stalk. The patient underwent transnasal-transsphenoidal surgery to remove the lesion. Histopathological findings revealed mixed inflammatory cells composed mainly of lymphocytes, macrophages and extensive fibrosis. CONCLUSIONS: Inflammatory pseudotumor of the pituitary, although rare, should be included in the differential diagnosis of a sellar and suprasellar mass. PMID- 15336879 TI - Transethmoidal approach to the optic canal: surgical and radiological microanatomy. AB - BACKGROUND: The purpose of the study is to describe anatomic topographic landmarks for transethmoidal approach to optic canal for optic nerve decompression. The study focuses on microsurgical/radiologic anatomies and their relationships in the region of the optic canal and orbit. METHODS: Human optic canal and related anatomic structures were studied in orbits of 6 formalin preserved adult human cadavers. In addition, anatomic measurements were made with digital vernier caliper on the orbits of 25 adult human skulls. The relation between ethmoidal and sinus was assessed with computed tomography (CT) scan in 25 living human heads needing cranial CT scan for any reason. RESULTS: The suture on the conjunction of frontal, maxilla, and lacrimal bones with a location in medial side of the orbit was accepted as a landmark. When the measurements were taken from this landmark, the distances to supraorbital margin were: right(R): 16.76 +/ 2.62 mm, left (L): 17.10 +/- 1.97 mm, and to infraorbital margin were R: 20.18 +/- 3.24 mm, L: 18.94 +/- 2.19 mm. The distances to the anterior ethmoidal foramen were R: 19.66 +/- 3.96 mm, L: 19.11 +/- 2.84 mm, and to the posterior ethmoidal foramen were R: 32.01 +/- 2.90 mm, L: 32.62 +/- 3.33 mm. Mean distance between the anterior and posterior ethmoidal foramen were R: 12.55 +/- 3.4 mm, L: 13.51 +/- 4.2 mm. The posterior ethmoidal foramen and optic ring were separated only by the mean distances of R: 5.34 +/- 2.81 mm, L: 4.9 +/- 3.35 mm. The distance from the suture to the distal (orbital) opening of the optic canal was R: 37.35 +/- 2.73 mm, L: 37.52 +/- 3.47 mm and to proximal (intracranial) opening of the canal were R: 49.52 +/- 2.62 mm, L: 50.94 +/- 3.35 mm. The average widths of proximal (intracranial) canal measured were R: 7.43 +/- 1.95 mm, L: 7.38 +/- 2.01 mm and those of distal canal (orbital) were R: 5.12 +/- 1.1 mm, L: 4.95 +/- 1.32 mm. The mean lengths of the optic canal were R: 11.19 +/- 2.68 mm, L: 12.42 +/- 3.38 mm. In radiologic examinations, the mean numbers of anterior group ethmoidal cells were R: 7, L: 6 and those of posterior group ethmoidal cells were R: 4, L: 3. The results of CT demonstrated 7 (14%) Onodi or sphenoethmoidal cells in 50 orbits of living humans. CONCLUSION: The examination of radiologic anatomy in addition to microanatomy can significantly contribute to preoperative and postoperative evaluation of the patients. PMID- 15336881 TI - Socialized medicine in Cuba part II: "Doctor diplomacy," sex tourism, and medical apartheid. PMID- 15336882 TI - Wrong disc space level surgery: medicolegal implications. PMID- 15336884 TI - Proximal nerve root spinal hemangioblastomas: presentation of three cases, MR appearance, and literature review. PMID- 15336886 TI - Heavy metals in recovered fines from construction and demolition debris recycling facilities in Florida. AB - A major product recovered from the processing and recycling of construction and demolition (C&D) debris is screened soil, also referred to as fines. A proposed reuse option for C&D debris fines is fill material, typically in construction projects as a substitute for natural soil. Waste material that is reused in a manner similar to soil must first be characterized to examine potential risks to human health and the environment. In Florida, samples of C&D debris fines from 13 C&D debris recycling facilities were characterized for 11 total and leachable heavy metal concentrations. Total metal concentration results (mg/kg) were compared to existing data on background Florida soil concentrations and to Florida's risk-based soil cleanup target levels (SCTLs). All of the detected metals were found to be elevated with respect to background. The 95% upper confidence level of arsenic from 99 samples was 3.2 mg/kg; arsenic presented the greatest limitation to reuse when compared to the SCTLs. Lead was not found to pose a major problem, likely because of the relatively new building infrastructure in Florida, which results in less demolition debris and less material impacted by lead-based paint. The results of batch leaching tests conducted using simulated rainwater (mg/l) were compared directly to risk-based groundwater levels for Florida and were found not to pose a risk using existing risk assessment policies. PMID- 15336887 TI - Setting action levels for drinking water: are we protecting our health or our economy (or our backs!)? AB - Clean and healthy drinking water is important for life. Drinking water can be drawn from streams, lakes and rivers, directly collected (and stored) from rain, acquired by desalination of ocean water and melting of ice or it can be extracted from groundwater resources. Groundwater may reach the earth's surface in the form of springs or can be extracted via dug or drilled wells; it also contributes significantly to river baseflow. Different water quality issues have to be faced when utilising these different water resources. Some of these are at present largely neglected in water quality regulations. This paper focuses on the inorganic chemical quality of natural groundwater. Possible health effects, the problems of setting meaningful action levels or maximum admissible concentrations (MAC-values) for drinking water, and potential shortcomings in current legislation are discussed. An approach to setting action levels based on transparency, toxicological risk assessment, completeness, and identifiable responsibility is suggested. PMID- 15336888 TI - Quantitative assessment of the effects of agricultural practices designed to reduce 137Cs and 90Sr soil-plant transfer in meadows. AB - Agricultural practices (ploughing and reseeding, addition of lime and fertiliser) were tested as a feasible remediation strategy to reduce 137Cs (RCs) and 90Sr (RSr) soil-plant transfer in natural meadows in areas affected by the Chernobyl fallout. Field experiments were carried out for 2 years at six sites, covering dry and wet meadows. Observed results at field scale showed that ploughing plus reseeding provoked the main reduction in RSr transfer, with no further reduction after liming, while ploughing + reseeding + K fertiliser led to the maximum decrease in RCs transfer at most sites. The direct effects of agricultural practices on the exchange complex and soil solution composition were quantified by subsequent soil analyses. At the doses applied, lime did not affect the Ca + Mg concentrations in the exchange complex and soil solution of the ploughed soils, thus suggesting that the decrease in RSr transfer on treated plots was mainly due to the changes in the plant species after reseeding. With respect to RCs, changes in the K+NH4+ concentrations in the exchange complex and soil solution were consistent with changes in soil-plant transfer. Finally, RSr and RCs soil-plant transfer in ploughed plots was well predicted from soil properties, such as the solid-liquid distribution coefficient, the ionic composition of the soil solution and the exchangeable cations, with Pearson correlation coefficients of 0.98 and 0.86, respectively, between calculated and experimental log transfer factors. PMID- 15336889 TI - Comparative studies of atmospheric trace elements: improvements in air quality near a copper smelter. AB - Measurements of trace element concentrations, including heavy metals in the atmosphere near a copper smelter in Port Kembla, New South Wales, have shown improvements in air quality, attributed mainly to modernisation of the plant, with more than 85% decrease (for each element) in airborne Cr, Zn, Se, Ag, Cd, Sb, Au and Pb, associated with 74% reduction in Cu concentration. This data was obtained by analysis of air particulate sampled in 2001-2002 for comparison with measurements at the same location in 1978. As expected, measurements of Na and Cl derived from the marine aerosol at this coastal location showed no corresponding change. The recent analysis of air particulate shows significant correlations between the monthly concentrations of Cu, Zn, Se, Ag and Pb, associated with industry, hence giving a high enrichment in airborne particulate by reference to the element/Sc ratios in 'average' soil. These enrichments, shown by heavy metals in particular, were two orders of magnitude greater than the values recorded in the atmosphere of rural areas; however, it is clear that air quality at Port Kembla is much improved and predominantly related to the modernisation of the smelter. PMID- 15336890 TI - Degradation of pentachlorophenol in cyclodextrin extraction effluent using a photocatalytic process. AB - This work evaluates a process for the elimination of pentachlorophenol (PCP) from effluents provided by a cyclodextrin-assisted flushing of contaminated soils. The effectiveness of photocatalytic degradation of PCP in several cyclodextrin (CD) solutions was evaluated using TiO2 as a photocatalyst. Effects of CD type on PCP degradation rate were studied at two pH values. A similar effect was observed for all CDs used on degradation rate of PCP and the decay of PCP was found to be less extensive at pH 11 than at pH 7. The kinetic orders of the photocatalytic reactions of PCP for all of the solutions have been determined. The first-order rate constants were found to be 0.0884, 0.0362, 0.0197 and 0.0053 min(-1) in CD solutions, respectively, at 0, 1, 2 and 5 mmol l(-1) of CD. Batch experiments were performed in order to study the CD extraction enhancement of PCP previously adsorbed on soil. The results show that the removal capacity of PCP from soil increases with CD concentration (from 0 to 5 mmol l(-1)). When the CD concentration was 5 mmol l(-1), an extraction of about 70% of PCP adsorbed on soil was observed, whereas only 37% was removed when water was used as the flushing solution. The optimal conditions for such a coupled method depend on the nature and concentration of the extracting agent and also on the photocatalytic experimental conditions. This work revealed that the coupling of cyclodextrin enhanced solubilization and photocatalytic treatment is a promising method for contaminated soil remediation. PMID- 15336891 TI - Absorption of foliar-applied arsenic by the arsenic hyperaccumulating fern (Pteris vittata L.). AB - The fact that heavy metals can enter various domains of the plant system through foliar pathways spurred us to explore if the fronds of the Chinese brake fern (Pteris vittata L.), a hyperaccumulator of arsenic, a carcinogenic metalloid, was proficient in absorbing arsenic in the form of sprays. The specific objective of this study was to investigate the impact of frond age, form of arsenic, and time of application on the absorption of foliar-applied arsenic by the brake fern; also examined were the effects of foliar sprays on surface ultrastructure and arsenic speciation in the frond following absorption. Foliar sprays of different arsenic concentrations (0, 50, 100, 200, and 400 ppm) were applied to young and fertile fronds. A positive linear relationship existed between arsenic concentration and absorption; the arsenic concentration of fronds increased from 50 to 200 ppm. Time-course analysis with excised pinnae indicated an initial linear increase followed by a plateau at 48 h. The young fronds with immature sori absorbed more arsenic (3100 ppm) than the fertile mature fronds (890 ppm). In the frond, the arsenic absorption was greatest in the lamina of the pinnae followed by the sori and the rachis. Applying arsenic during night (20:00-22:00 h) or afternoon (12:00-14:00 h) resulted in greater absorption of arsenic than the application in the morning (08:00-10:00 h). The arsenic absorption was greater through abaxial surfaces than through adaxial surfaces. The brake fern absorbed more arsenic when it was applied in the form of arsenite. Regardless of the form of arsenic and the surface it was applied to, arsenic occurred as arsenite, the reduced and the most toxic form of arsenic, after having been absorbed by the fronds. Scanning electron microscopy revealed no surface morphological alterations following all arsenic sprays. The study unequivocally illustrated that the Chinese brake fern absorbed foliar-applied arsenic with great efficiency. Consequently, the arsenic concentrations in the fronds transcended the levels of hyperaccumulation; such a characteristic could be exploited in the phytoremediation of groundwater contaminated with arsenic. PMID- 15336892 TI - Twenty-four-hour urinary excretion of ten pesticide metabolites in healthy adults in two different areas of Italy (Florence and Ragusa). AB - The determination of pesticide metabolites in human biological fluids represents an important biomarker of exposure in the general population and exposed workers. In the frame of a prospective study, we measured the 24-h urinary excretion of 10 pesticide metabolites to evaluate non-occupational exposure to pesticides in the general population in two different areas in Italy. We collected 24-h urine samples from 69 healthy adults residing in Florence (Central Italy, n = 51) and Ragusa (Southern Italy, n = 18). The volunteers (25 males, 44 females; mean age 56 years) did not report any occupational exposure. We measured: six alkylphosphates, aspecific metabolites of organophosphorus pesticides [dimethylphosfate (DMP), dimethylthiophosfate (DMTP), dimethyldithiophosfate (DMDTP), diethylphosfate (DEP), diethylthiophosfate (DEDP), and diethyldithiophosfate (DEDTP)]; 3,5,6-trichloro-2-pyridinol (TCP), the main metabolite of chlorpyrifos; 3-phenoxybenzoic acid (3-PBA), a metabolite of pyrethroid insecticides; ethylenethiourea (ETU) a metabolite of ethylenebisdithiocarbamates; methamidophos (METH), an organophosphorus insecticide. We also measured PABA excretion as compliance marker (mean recovery 95%). Dimethylphosphates were found in detectable concentrations in the majority of samples (89.9%, 82.6% and 60.9% for DMP, DMTP and DMDTP, respectively). Urinary diethylphosphates (DEP, DETP, DEDTP) concentrations were above the detection limit in 80.9%, 61.8% and 27.5% of samples, respectively. TCP, 3-PBA and ETU were detected in 78.3%, 53.6% and 21.7% of samples, respectively. Methamidophos was detected in two samples (2.8%). The median number of metabolites detected in the same urine sample was 6 (range 0-9). Excretion levels were highest for alkylphosphates, particularly for DMTP (median: 142.6 nmol/day). Multivariate analysis showed statistically significant differences between these two groups of adults, with higher mean values of urinary excretion of alkylphosphates, TCP and ETU in Florence in comparison to Ragusa. Overall, a very high percentage of 24-h urine samples positive for several pesticide metabolites emerged, with higher levels of urinary daily excretion in subjects residing in the more urbanised area. Our results suggest that food monitoring programs should be supported by general campaigns aimed to reduce the use of pesticides in agriculture. PMID- 15336893 TI - A simple approach to performing quantitative cancer risk assessment using published results from occupational epidemiology studies. AB - Quantitative risk assessment provides formalized scientific input for setting occupational and environmental standards for potentially toxic exposures. We describe the methods to perform quantitative cancer risk assessment based on published epidemiological literature using a hand calculator or spreadsheet. These methods are illustrated for some suspected or known human carcinogens, including perchloroethylene, formaldehyde, benzene and nickel. The approach to cancer risk assessment presented here may prove useful for those without access to original data from epidemiologic studies. The methods described should facilitate the use of published epidemiological data in the estimation of cancer risk in the general or working population. PMID- 15336894 TI - Concentrations of organotin compounds and imposex in the gastropod Hexaplex trunculus from the Lagoon of Venice. AB - In four stations located close to the channels connecting the Lagoon of Venice with the Adriatic Sea (two inside the lagoon and two outside it), individuals of Hexaplex trunculus were collected in order to assess their contamination. Concentrations of some organotin compounds, i.e. tributyltin and triphenyltin with their di- and mono- substituted metabolites, were measured and endocrine disruption such as imposex (superimposition of male sexual characteristics on females of gonochoristic gastropods) was observed. Vas deferens sequence indexes (VDSI) of 4.1-4.9 were found in organisms from stations inside the lagoon, and 3.6-4.2 in the sea stations. Organotin derivatives were measured in both the visceral coil and the rest of the soft body of the organisms. Total concentrations of butyltin compounds ranged from 102 +/- 17 to 432 +/- 27 ng Sn g(-1) d.w. in the visceral coil, and from 96 +/- 24 to 297 +/- 107 ng Sn g(-1) d.w. in the rest of the soft body. Phenyltins were found at far lower concentrations, ranging from 8 +/- 1 to 41 +/- 3 ng Sn g(-1) d.w. (visceral coil) and from 0.25 to 32 +/- 22 ng Sn g(-1) d.w. (rest of soft body). The degree of imposex in female gastropods, evaluated from VDSI and penis lengths, was related to organotin contents in the soft body. In particular, female penis lengths were significantly correlated (r = 0.917 and r = 0.982, P<0.05) to tributyltin (TBT) contents and the sum of organotin compounds in organisms. PMID- 15336895 TI - Prenatal and postnatal exposure to environmental pollutants in sewage sludge alters emotional reactivity and exploratory behaviour in sheep. AB - In order to find out whether exposure to environmental pollutants (EP) present in sewage sludge can change the behaviour of sheep, we compared the behaviour of two groups of 5-month old lambs (Ovis aries) with respect to their emotional reactivity and exploratory behaviour. One group (treated, T) comprised the offspring of ewes who had been kept throughout their lives on pastures with slightly elevated, environmental levels of pollutants, as a result of the application of sewage sludge at rates used in normal practice. The other group (control, C) were the offspring of ewes whose pastures had been treated with inorganic fertiliser. During a 1-min period in the weigh crate, T lambs were less reactive than C lambs, but vocalised more. Exploratory behaviour showed a clear sex difference, with female C lambs exploring more than male C lambs. However, this difference was not found in the T lambs, where both males and females showed high levels of exploration. This points to a demasculinising effect of exposure to higher pollutant concentrations with respect to exploratory behaviour. These observations demonstrate the need to take account of the effects of combinations of pollutants, even at very low, environmental concentrations, and further highlight the usefulness of ethotoxicology for the study of biological effects of environmental pollutants. PMID- 15336896 TI - Water quality characteristics of vegetated groundwater-fed ditches in a riparian peatland. AB - The Environmental Ministry of Schleswig-Holstein (Northern Germany) has implemented a novel peatland rehabilitation programme to utilise the high denitrification potential of degenerated, minerotrophic peatlands for the reduction of nitrate input into aquatic ecosystems, and to simultaneously improve the habitat conditions. Realisation of both goals requires changes to the water management adapted to the site-specific geohydrological conditions. The effect of raised groundwater levels and extensive land use on the water quality of heavily vegetated and groundwater-fed ditches was investigated in a riparian peatland located in the River Eider Valley (a nationally important wetland case study area). The water quality of the selected representative ditches was regularly assessed at different discharge levels during different flow obstruction cover periods. The hydraulic residence time was predominantly a function of the ditch geometry and the overall flow obstruction. A better understanding of the effect of ditch vegetation on the temporal flow patterns and the hydraulic residence times is of high environmental interest, especially for improving nutrient standards in lowland rivers such as the River Eider. Within-ditch vegetation and other hydraulic flow obstructions such as accumulated silt and organic debris increased the hydraulic residence time and led to an improvement of the water quality (e.g. reduction in nitrate content) along the ditch. While ortho phosphate and ammonia concentrations were acceptable to German water quality standards, nitrate-nitrogen concentrations were frequently elevated due to high discharges despite high flow obstruction cover. Further findings show that the lower stretches of the ditches were flooded by the River Eider due to the absence of a macrophyte-mowing scheme that led to increased water levels during late summer. PMID- 15336897 TI - The role of climate variability in the inter-annual variation of terrestrial net primary production (NPP). AB - Eleven years data set of global net primary production (NPP) and long-term climatic and land use data were used to explore the patterns of inter-annual variability of terrestrial NPP in relation to potential causal factors. Global anomalies in temperature, precipitation and cloud cover were found to significantly contribute in different ways and magnitudes to the variability of NPP of global ecosystems particularly forests and grasslands. El Nino/La Nina events represented an important factor affecting forests, woodlands and grasslands while deforestation was found to largely contribute to the NPP variability of tropical forests. Regionally, NPP variability is related to variation of precipitation in the tropics but is related to both variation and annual mean of temperature and cloud cover in the mid-northern latitudes. We hypothesized that the increase in variability of potential causal factor(s) will provoke more declines of NPP in the tropics but will yield more pulses or at least maintain a mean NPP in the mid-northern latitudes. PMID- 15336898 TI - Introduction to the DAPPLE Air Pollution Project. AB - The Dispersion of Air Pollution and its Penetration into the Local Environment (DAPPLE) project brings together a multidisciplinary research group that is undertaking field measurements, wind tunnel modelling and computer simulations in order to provide better understanding of the physical processes affecting street and neighbourhood-scale flow of air, traffic and people, and their corresponding interactions with the dispersion of pollutants at street canyon intersections. The street canyon intersection is of interest as it provides the basic case study to demonstrate most of the factors that will apply in a wide range of urban situations. The aims of this paper are to introduce the background of the DAPPLE project, the study design and methodology for data collection, some preliminary results from the first field campaign in central London (28 April-24 May 2003) and the future for this work. Updated information and contact details are available on the web site at http://www.dapple.org.uk. PMID- 15336899 TI - Organophosphorus flame retardants and plasticisers in surface waters. AB - Organophosphates used as flame retardants, plasticisers and lubricants such as tris-(2-chloro-, 1-methyl-ethyl)-phosphate (TCPP), tris-(2-chloroethyl)-phosphate (TCEP) or tris-(2-chloro-, 1-chloromethyl-ethyl)-phosphate (TDCP), tri-n butylphosphate (TnBP), tri-iso-butylphosphate (TiBP), triphenylphosphate (TPP) and tris-(butoxyethyl)-phosphate (TBEP) have been analysed in several rivers and sewage treatment plant (STP) effluents. The concentrations in the River Ruhr are 20-200 ng/l TCPP, 13-130 ng/l TCEP, about 50 ng/l TDCP, 10-200 ng/l TBEP and up to 40 ng/l TPP. The STP effluents exhibit concentrations up to 400 ng/l TCPP, 130 ng/l TCEP, about 120 ng/l TDCP and 500 ng/l TBEP, respectively. The main sources for the load of organophosphates are sewage treatment plants, but not all contribute equivalent to the amount of inhabitants they serve. PMID- 15336900 TI - Concentration and fate of trace metals in Mekong River delta. AB - In the Mekong River delta and its associated coastal zone trace elements concentrations (Cd, Cu, Ni and Pb) were measured in the dissolved phase (DP) during dry (March 1997) and wet (October 1997) seasons. As, Co, Cr, Ni, Pb, and Al were also measured in suspended matter (SM) and total and organic carbon, trace elements (Cd, Cr, Cu, Hg, Mn, Ni, Pb, Zn) and macro elements in superficial sediments (S). Trace metal concentrations in DP and SM during the contrasting hydrological conditions were generally found within the range observed for uncontaminated environment. The average DP concentrations (nM) in the river for March and October are: Cd 0.03 and 0.09, Cu 15 and 14, Ni 7.8 and 8.4, Pb 0.51 and 0.50, respectively. In general there is no significant difference between the concentrations observed during dry and wet season. The evolution of the DP trace metal concentration in the surface water within the salinity gradient suggests no noticeable exchange between the particulate and dissolved phase. This result is in good agreement with those observed in most plume structures studied so far. The average concentrations in the SM (microg/g) (March, October) at the river end member are: As (24; 11), Co (17; 9), Cr (49; 29), Ni (32; 18), Pb (42; 19) and Al (113,000; 67,000), respectively. All trace elements show higher concentrations in March than in October, with an average increase of two times. This is essentially related to grain size effect since smaller particles were supplied during dry season. These differences are not reflected in the mixing zone, which integrates the seasonal variations. The concentration of major elements (C total, C organic, Si, Al, Ca, K, Fe, Mg, Ti), trace elements (Pb, Zn, Cu, Ni, Mn, Cr, Cd, Hg) in superficial sediments, show similar values during the two seasons and does not show any important variation with depth, indicating either a very fast sedimentation rate and/or the absence of any significant contamination. PMID- 15336901 TI - Partitioning and desorption behavior of polycyclic aromatic hydrocarbons from disparate sources. AB - Contaminated sediments pose a unique challenge for risk assessment or remediation because the overlying water column may transport contaminants offsite or to ecological receptors. This research compares the behavior of polycyclic aromatic hydrocarbons (PAHs) on marine sediments from two sites. The first site was affected by shipping activities and the second was impacted by a creosote seep. Organic carbon:water partitioning coefficients (Koc values) were measured with three solutions. Desorption was measured using Tenax beads. PAHs from the ship channel had lower Koc values than those from the creosote facility. For example, the average logKoc value of ship channel pyrene was significantly lower than that of creosote facility pyrene (4.39 +/- 0.35 and 5.29 +/- 0.09, respectively, when tested in 5 mM calcium chloride). These results were consistent with the greater desorption of pyrene, phenanthrene and benzo(a)pyrene from the ship channel than from the creosote facility sediments. Organic compound desorption from sediments can be considered to be a two-stage process, with a labile fraction that desorbs quickly and a refractory fraction that desorbs much more slowly. In both sediments, more than 75% of the benzo(a)pyrene was found to have partitioned into the refractory phase. The amounts of phenanthrene and pyrene that partitioned into the refractory phase were lower. Linear correlations of logKoc with log(CR/CL) (where CR and CL are the fractions of the compound in the refractory and labile phases, respectively, at time zero) showed that partitioning measurements made with the US EPA's Toxicity Characteristic Leaching Procedure fluid (US EPA, 1996) most closely matched predictions of desorption behavior. The data imply that with a larger data set, it may be possible to relate simple partitioning measurements to desorption behavior. Partitioning measurements were used to predict water concentrations. Despite having higher concentrations of carcinogenic PAHs [cPAHs, the seven PAHs categorized by the US EPA (2004) as class B2 carcinogens], creosote facility sediments were predicted to produce lower aqueous concentrations of cPAHs. These results indicate that both sediment and contaminant characteristics will impact contaminant release from sediments. PMID- 15336902 TI - Washout of accumulated testosterone in a watershed. AB - Testosterone is constantly excreted into the environment by both human and animal sources but little is known about how it is transported in the environment. In this study, testosterone was measured in 15 sites in the Upper Jordan Valley after major rain events (238 samplings) for two consecutive rain seasons. The area consists of small farms, cattle pasture, fish ponds with some urban development. One liter samples were extracted on solid phase columns and the eluates measured using specific radio-immunoassay for testosterone and estrogen (estradiol + estrone). The first rain season was the first above average season after a 3-year period of well below average rainfall. It was found in the rain season of 2001/2002, that following a rain sequence of 131 mm/week there was an initial large increase in the concentration of testosterone (maximum 6 ng/l) accompanied by high estrogen (maximum 6 ng/l), which then gradually declined to non-detectable levels (< 0.3 ng/l) over a period of 3 months. These peaks originated from runoff from cattle pasture and fish pond effluent. Later peaks consisted only of testosterone that was moderately associated with sulfate (r2 = 0.53, P < 0.05) and somewhat associated with total phosphorus (r2 = 0.49, P < 0.1) indicating that the origin was leaching from the sulfurous peat soil. In the following rainy season, which had recorded rainfalls, no testosterone peaks above 1 ng/l were seen. We conclude that the testosterone accumulated in the Upper Jordan Valley was washed out in two stages, first as surface runoff from cattle pasture and then as discharge from the soil. PMID- 15336903 TI - Solid-solution partitioning of plutonium in surface waters at the Atomic Weapons Establishment Aldermaston (UK). AB - The Atomic Weapons Establishment (AWE) at Aldermaston (Berkshire, UK) has provided and maintained the warheads for the UK's nuclear deterrent for more than 50 years. Whilst the site is radiologically safe, in a few locations the soil contains specific activities of plutonium (Pu) above background arising from a legacy of historic operations. Run-off water (a mixture of rainwater and groundwater) from part of the site is routed into a water management system, and after analysis and radiological assessment, released into local streams. Water and sediment samples have been collected from a number of closely spaced locations within this system to assess the solid-solution partitioning of Pu. Survey work was complemented by batch type desorption experiments to assess redissolution from 'contaminated' sediment into 'uncontaminated' water. The survey data indicate that specific activities of both dissolved and particle bound 239 + 240Pu varied by roughly two orders of magnitude, ranging from approximately 0.7 microBq kg(-1) up to approximately 44 microBq kg(-1), and approximately 1.2 Bq kg(-1) up to approximately 400 Bq kg(-1), respectively, consistent with water originating from different parts of the site. Apparent Kd values varied by an order of magnitude (from 0.7-16 x 10(6)) with an average value of 4 x 10(6). Results from the desorption experiments indicated the extent of redissolution was very small and the derived Kd's corroborated values obtained from the survey work. Kd's given here are compared with other literature values, and are the greatest reported to date. Results are also provided describing the variation in water quality parameters in shallow groundwater samples. Alkalinity values ranged from 120 to 388 mg l(-1) CaCO3 with an average value of 195 mg l( 1) CaCO3. Corresponding values for pH were 6.6-8.3 with an average of 7.5. Over half of the samples were estimated to be supersaturated with respect to calcite. It is suggested that the state of calcite saturation may be responsible for the high Pu Kd. As a consequence of the high particle reactivity, migration of Pu contamination, both within and away from the AWE Aldermaston site, is likely to be very restricted. PMID- 15336904 TI - Spatial variation in nitrogen dioxide in three European areas. AB - In order to estimate the spatial variation within well-defined study areas, nitrogen dioxide was measured with diffusion samplers (Palmes tube) in 40-42 sites each in Germany (Munich), the Netherlands and Sweden (Stockholm County). Each site was measured over four 2-week periods during 1 year (spring 1999 to summer 2000). In each country, one reference site was measured during all periods and the results were used to adjust for seasonal variability, to improve the estimates of the annual average. Comparisons between the chemiluminescence method (European reference method) and Palmes tube measurement indicated a good agreement in Germany (with a ratio of 1.0 for Palmes tube/chemiluminescence) but underestimation for Palmes tube measurement in the Netherlands and Sweden (0.8 for both countries). The r2 values were between 0.86 and 0.90 for all three countries. The annual average values for NO2 for different sampling sites were between 15.9 and 50.6 (mean 28.8 microg/m3) in Germany, between 12.1 and 50.8 (mean 28.9 microg/m3) in the Netherlands and between 6.1 and 44.7 (mean 18.5 microg/m3) in Sweden. Comparing spatial variation between similar sites in the three countries, we did not find any significant differences between annual average levels for urban traffic sites. In Sweden, annual average levels in urban background and suburban backgrounds sites were about 8 microg/m3 lower than comparable sites in Germany and the Netherlands. Comparing site types within each country only urban traffic sites and suburban background sites differed in Germany. In the Netherlands and Sweden, the urban traffic sites differed from all other sites and in Sweden also the urban background sites differed from the other background sites. The observed contribution from local traffic was similar in the Netherlands and Sweden (10 and 8 microg/m3, corresponding to 26-27% of the NO2 concentration found in the urban traffic sites). In Germany, the contribution from local traffic was only 3 microg/m3, corresponding to 9% of the NO2 concentration found in the urban traffic sites. The spatial variation was substantially larger for NO2 than the variation for PM2.5 and similar to PM2.5 absorbance, measured in the same locations. PMID- 15336905 TI - Biogeochemistry of organotin compounds and tin in a forested catchment in Germany. AB - Organotin compounds (OTC) are highly toxic pollutants that have been shown to affect many aquatic ecosystems. Little is known about the input and fate of OTC in terrestrial ecosystems. Here, soil pools, concentrations and fluxes in bulk precipitation, throughfall, fog, litterfall and runoff of OTC and Sntotal were investigated in a forested ecosystem (Picea abies, Karst.) in NE Bavaria, Germany. The concentrations of OTC and Sntotal were generally in the order fog>throughfall>bulk precipitation. Average concentrations of OTCtotal ranged from 57 ng Sn l(-1) in fog to 5.8 ng Sn l(-1) in bulk precipitation. Concentrations of Sntotal were in the same order but between 490 ng Sn l(-1) in fog and 140 ng Sn l(-1) in bulk precipitation, on average. Average OTCtotal concentrations in litterfall were 12.9 ng Sn g(-1) and those of Sntotal in litterfall 38 ng Sn g(-1). All OTC concentrations in runoff were lower than in bulk precipitation, while those of Sntotal were similar to the concentrations in bulk precipitation. Monobutyltin was the dominating OTC in bulk precipitation, throughfall, fog and litterfall, but was seldom detected in the runoff. The annual total deposition of OTCtotal (calculated as throughfall + litterfall) was 172 mg Sn ha(-1) year(-1), with 45 mg Sn ha(-1) year(-1) represented by litterfall. The annual runoff from the catchment of OTCtotal amounted to 25 mg Sn ha(-1) year(-1). The total deposition of Sntotal was 4.9 g Sn ha(-1) year(-1), of which 0.2 g Sn ha(-1) year(-1) was litterfall. The annual runoff of Sntotal was 2.4 g Sn ha(-1) year(-1). The mass balance showed a high retention of OTC and Sntotal in the catchment. The forest soils act as a strong sink for OTC and Sntotal. Only small amounts of deposited OTC are released to runoff. The ratio of soil pools to annual accumulation for total OTC (46 years) indicates that OTC inputs have been occurring already for many decades or have been substantially higher in the past than today. PMID- 15336906 TI - Wet and dry deposition patterns of plutonium in Daejeon, Korea. AB - Plutonium in wet and dry deposition samples collected monthly at the Korea Institute of Nuclear Safety (KINS), Daejeon, Korea was determined during the period from January 2000 to August 2000. Monthly 239,240Pu deposition in Korea showed a maximum in the spring season (March to May), which was several times greater than that in Japan and corresponds to a seasonal cycle of soil dust fallout originating from the East Asian arid areas. The trajectory analysis of dust storms suggests that 239,240Pu deposition in Korea in spring is originating from plutonium-bearing surface soil particles from the East Asian arid areas. A significant part of the 239,240Pu deposition in spring in Korea is attributable to dry deposition. PMID- 15336907 TI - Determination of biodegradable dissolved organic carbon in waters: comparison of batch methods. AB - An effect of different types of bacterial inocula upon the final biodegradable dissolved organic carbon (BDOC) result was investigated in samples of both low and high BDOC concentrations. Stream water and leaf leachate samples were incubated either with free, suspended bacteria or with bacteria attached to the stream sediment particles or attached to artificial substrata. The time course of dissolved organic carbon (DOC) decomposition was observed using absorbance analysis of DOC. BDOC determination by means of commonly used suspended bacteria as the inoculum made for an underestimation of BDOC between 5% and 25%, compared with attached bacterial community (biofilm). The reason for these findings could be the higher microbial diversity, higher metabolic activity of attached bacteria and abiotic adsorption of organic molecules to inorganic support and biofilm matrix surfaces. Adsorbed DOC is easily hydrolyzed and utilized by biofilm bacteria. PMID- 15336908 TI - On the logarithmic relationship between NO2 concentration and the distance from a highroad. AB - Measurements of nitrogen dioxide (NO2) concentrations, performed with passive diffusion samplers in gradients from a highway in South-west Sweden, were used to test the assumption that the NO2 concentration contributed by the highway varies with the logarithm of the distance from the highway. The five data sets used corroborated the hypothesis, and it was shown that all data could be accommodated to a common relationship with high correlation (R2=0.95) using the concentration of 10 m away from the highroad as the reference. The data were also well in accordance with a recently published study from Canada, although the slope of the relationship between the NO2 concentration contributed by a highway and the logarithm of the distance was somewhat stronger for the Swedish data compared to the Canadian. The regression slope is likely to be sensitive to wind speed, atmospheric stability, surface roughness and the background ozone concentrations of the area. PMID- 15336909 TI - The Werner syndrome protein at the crossroads of DNA repair and apoptosis. AB - Werner syndrome (WS) is a premature aging disease characterized by genetic instability. WS is caused by mutations in a gene encoding for a 160 kDa nuclear protein, the Werner syndrome protein (WRN), which has exonuclease and helicase activities. The mechanism whereby WRN controls genome stability and life span is not known. Over the last few years, WRN has become the focus of intense investigation by a growing number of scientists. The studies carried out by many laboratories have provided a wealth of new information about the functional properties of WRN and its cellular partners. This review focuses on recent findings that demonstrate a functional interaction between WRN and two factors that bind to DNA breaks, Ku and poly(ADP-ribose) polymerase 1, and discuss how these interactions can influence fundamental cellular processes such as DNA repair, apoptosis and possibly regulate cell senescence and organismal aging. PMID- 15336910 TI - Effect of ageing and caloric restriction on specific markers of protein oxidative damage and membrane peroxidizability in rat liver mitochondria. AB - To gain insight into the anti-ageing mechanisms of caloric restriction (CR), liver mitochondria were isolated from male Brown-Norway rats of different ages (fully fed control and CR) and various specific markers of non-enzymatic protein modification (by oxidative, glyco- and lipoxidative-reactions) were measured by GC/MS and Western blotting. A membrane peroxidizability index (PI) was calculated from the fatty acid profiles. Between 6 and 18 months of age, there were significant decreases in the concentration of all markers of damage in mitochondria from both the fully fed and CR groups. In contrast, between the ages of 18 and 28 months, there were significant increases in the concentrations of all markers of damage. In mitochondria from both fully fed and CR groups, there were significant increases in N-epsilon (Nepsilon)-(carboxymethyl)lysine (CML) and N-epsilon-(malondialdehyde)lysine (MDAL) between 6 and 28 months of age. In general, damage tended to be lower in mitochondria from CR animals, but the effects were not significant, except for the concentration of N-epsilon (carboxymethyl)lysine at 28 months of age. PI increased steadily and significantly with age in fully fed animals, whilst CR induced a significant decrease in this index at 28 months of age. It is concluded that for male rats of the Brown-Norway strain, and mitochondria from liver (i) old (but not mature) age is associated with an increased membrane PI and protein oxidative damage and (ii) CR does not lead to a general reversion in age-related protein damage, but it does prevent the age-induced increase in PI very late in life. PMID- 15336911 TI - Role of defective ERK phosphorylation in the impaired GM-CSF-induced oxidative response of neutrophils in elderly humans. AB - GM-CSF-induced oxidative responses are defective in neutrophils of elderly humans. In the present study we evaluated whether this phenomenon might be related to alterations in cytokine-dependent MAPK signalling. Neutrophils obtained from elderly humans and stimulated with GM-CSF showed a significant reduction in phosphorylated ERK1/2 levels and an even higher decrease in ERK1/2 activation with respect to baseline. No changes in GM-CSF-induced p38 MAPK phosphorylation were observed. Cell pretreatment with the MEK inhibitor PD98059 determined a marked suppression of GM-CSF-induced O2- release. Interestingly, under the above experimental condition, there was no longer any difference in O2- production observed between elderly and young subjects. Furthermore, despite the fact that the p38 MAPK pathway was activated less strongly by GM-CSF, the p38 MAPK inhibitor SB203580 reduced GM-CSF-induced O2- production in the neutrophils of the elderly to levels similar to those obtained with PD98059. TNF-alpha triggered O2- production was not altered by ageing and in fact, a similar ERK1/2 or p38 MAPK activation was found in TNF-alpha-stimulated neutrophils from elderly and young individuals. In accordance with the different potency of TNF-alpha in activating ERK1/2 and p38 MAPK, the TNF-alpha-induced oxidative responses were more sensitive to the inhibitory effects of SB203580 than to those of PD98059 in young as well as elderly subjects. These results suggest that, along the GM-CSF dependent ERK signalling pathway, a step proximal to MEK1/2 but distal to the connection with the p38 MAPK module likely becomes defective as a feature of age. The consequent decline in ERK1/2 activation could potentially account for the GM CSF-dependent impairment of the neutrophil respiratory burst that occurs with ageing. PMID- 15336912 TI - Developments of geriatric autopsy database and Internet-based database of Japanese single nucleotide polymorphisms for geriatric research (JG-SNP). AB - To facilitate geriatric research on the roles of genetic polymorphisms of candidate genes, two databases were developed based on data obtained from autopsy examinations of elderly subjects: the geriatric autopsy database (GEAD) and the Japanese single nucleotide polymorphisms (SNP) database for geriatric research (JG-SNP) which is accessible on the Internet (http://www.tmgh.metro.tokyo.jp/jg snp/english/E_top.html). The data for the GEAD were derived from 1074 consecutive autopsy cases (565 male and 509 female cases) with an average age of 80 years. The GEAD was installed on a stand-alone Windows 2000 server using Oracle 8i as the database application. The GEAD contains clinical diagnoses of 26 geriatric diseases, histories of smoking and alcohol consumption, pathological findings (720 items), severity of atherosclerosis, genetic polymorphism data, etc. On the JG-SNP website, case distribution corresponding to a specified SNP or disease can be searched or downloaded. Although there are several Internet-based SNP databases such as dbSNP, no databases are available at present on the web that contain both SNP data and phenotypic data. As autopsy studies can provide large amounts of accurate medical information, including the presence of undiagnosed diseases such as latent cancers, the GEAD is a unique and excellent database for research on genetic polymorphisms. PMID- 15336913 TI - The decline of postnatal mortality from gastrointestinal anomalies with age. AB - It has been previously shown that mortality from congenital anomalies decreases with the first power of age with the exception of "gastrointestinal anomalies". On the other hand, mortality declines with the second power of age in the category "certain conditions originating in the perinatal period". According to the theory of congenital individual risks, the dying out of more impaired individuals causes these two types of mortality decline. Mortality decline with the first power of age is the most frequent type of mortality decline after birth. This evidence is explained as a consequence of the rule: the higher the individual risk of death, the less frequent the individual risk is at the moment of birth. If all defects (and all individual risks) appear with the same frequency and selection does not occur (e.g. this could be assumed for certain conditions originating in the perinatal period) then mortality declines with the square of age. Here, it is shown that mortality from "gastrointestinal anomalies" (ICD9 codes: 750 and 751) decreases with the second power of age in all five countries within the age interval 1-10 years. This is a consequence of evidence that these defects do not affect vitals important during pregnancy and selection is not significant. These findings demonstrate that the selection of higher individual risks proceeds during pregnancy rather than during previous generations. PMID- 15336914 TI - Human chromosomes with shorter telomeres and large heterochromatin regions have a higher frequency of acquired somatic cell aneuploidy. AB - Both telomere shortening and increases in aneuploidy frequencies have been associated with aging. To test if these chromosomal attributes are correlated, chromosome-specific telomere lengths and aneuploidy frequencies were estimated and compared. Aneuploidy frequencies were determined for 10 autosomes (1, 3, 5, 8, 9, 10, 13, 16, 17, 21) and the X chromosome in lymphocytes, and for chromosomes 17 and X in buccal mucosa cells. Overall, chromosomal loss was seen more often than gain in lymphocytes, with the highest loss rates being observed for chromosomes X (3.03%), 17 (2.00%), and the autosomes having large blocks of heterochromatin (1 [1.93%]; 16 [1.53%]; and 9 [1.05%]). The frequencies of loss were significantly lower in the buccal mucosa cells compared to lymphocytes for chromosomes 17 (P = 0.006) and X (P = 0.003). However, the chromosome 17 trisomy frequencies did not vary between tissues. Using a semi-quantitative FISH assay to estimate chromosome-specific telomere length, a significant negative correlation (r = -0.379; P = 0.007) was seen for chromosomal aneuploidy and telomere length, with chromosomes having higher loss rates being noted to have shorter telomeres. Collectively, these studies show that acquired, spontaneous chromosomal loss is associated with multiple factors including the amount of heterochromatin, the chromosome's telomere length, and tissue-specific factors. PMID- 15336915 TI - The concomitant presence of polymorphic alleles of interleukin-1beta, interleukin 6 and apolipoprotein E is associated with an increased risk of myocardial infarction in elderly men. Results from a pilot study. AB - Genetic background of inflammatory or anti-inflammatory molecules may be helpful in identifying subjects with increased or decrease risk of developing cardiovascular disease. Bi-allele polymorphism (C > T) in the promoter region ( 511) of the interleukin-1beta (IL-1beta) gene and the bi-allele polymorphism (G > C) in the promoter region (-174) of interleukin-6 (IL-6) gene were determined in elderly men patients with myocardial infarction (MI) and healthy controls. Each subject was also genotyped for the triallelic polymorphism of the apolipoprotein E epsilon gene. The IL-6C and APOE epsilon4 alleles were independently associated with a mild or moderate increased risk of MI, whilst the allele C of the IL-1beta was not independently linked to MI risk. However, the simultaneous presence of the allele C of IL-1beta, the allele C of IL-6 and epsilon4 allele of APOE was strongly associated with the disease. Data from this cross-sectional study suggest that the functional interaction of these three genes affects pathogenetic mechanisms of MI and an impaired regulation of immune responses plays a pivotal role in the disease. Furthermore, genetic background of inflammatory genes may influence longevity of human species by affecting inflammatory responses associated to cardiovascular diseases. The administration of anti-inflammatory compounds to middle age healthy subjects with increased genetic susceptibility of developing MI might decrease the incidence and prevalence of cardiovascular events in aging. PMID- 15336916 TI - Infants' sensitivity to correlations between static and dynamic features in a category context. AB - Four experiments with the habituation procedure investigated 14-22-month-olds' ability to attend to correlations between static and dynamic features embedded in a category context. In Experiment 1, infants were habituated to four objects that exhibited invariant relations between moving features and motion trajectory. Results revealed that 14-month-olds did not process any independent features, 18 month-olds processed individual features but not relations among features, and 22 month-olds processed relations among features. In Experiment 2, 14-month-olds differentiated all of the features in the events in a simpler discrimination task. In Experiments 3a and 3b, 22-month-olds failed to show sensitivity to correlations between dynamic and static features in a category context. In Experiment 4, 22-month-olds, but not 18-month-olds, generalized the learned feature-motion relation to a novel instance. The results are discussed in relation to infants' developing ability to attend to correlations, constraints on learning, category coherence, and the development of the animate-inanimate distinction. PMID- 15336917 TI - Inviting witnesses to speculate: effects of age and interaction on children's recall. AB - Inviting speculation has been found to increase children's false recall. In this study, kindergartners and third graders saw a clown perform actions alone or in interaction with a child. Two weeks later, the speculation group recalled all actions and was asked to speculate on half the actions. The control group recalled all actions without speculating. Four weeks after the show, all children recalled all actions again. The speculation group gave more false answers to the speculated items than the control group. Surprisingly, older children tended to report as many if not more false responses than younger children, regardless of speculation. In the speculation group, there were fewer false answers for interactions than actions, but false answers did not differ across observation types in the control group. Finally, speculation did not affect free and cued recall differentially. PMID- 15336918 TI - The effects of memory trace strength on eyewitness recall in children with and without intellectual disabilities. AB - Children with mild moderate intellectual disabilities (ID) were compared with typically developing peers of the same chronological age (CA) on an eyewitness memory task in which memory trace strength was manipulated to examine whether increased memory trace strength would benefit those with ID more than those without ID. No evidence was found for this claim or for the notion that different mechanisms are implicated in memory processes for children with ID versus CA controls. Fuzzy-trace theory was also used to contrast question types that probed verbatim memory versus gist memory. Manipulations of trace strength, when used with immediate recall (to reduce the impact of decay), were predicted to improve verbatim memory more than gist memory. The results broadly supported the predictions. Performance was not improved in the stronger trace strength condition on measures of recall that tapped gist memory (e.g., open-ended recall), whereas performance was significantly better in the stronger trace strength condition on two of the three measures of recall that tapped verbatim memory (i.e., closed misleading questions, open-ended specific questions). Differences in performance between the groups were quite marked on several question types, supporting previous findings that those with ID have certain vulnerabilities as potential witnesses compared with peers of the same CA. PMID- 15336919 TI - Selective search for conjunctively defined targets by children and young adults. AB - Two experiments were conducted to investigate age-related differences in visual search for targets defined by the conjunction of two features. In the experiments, 7- and 10-year-old children and young adults searched visual displays for a black circle among distractors consisting of gray circles and black squares. In Experiment 1 (N = 60), we compared performance in the standard search task (where an equal number of each type of distractor appeared across all display sizes) with performance in a modified search task (where the number of black squares was fixed at two and the number of gray circles increased as the display size increased). In Experiment 2 (N = 60), the ratio of black stimuli to gray stimuli was varied systematically as the display size increased. Results of both experiments indicated that all participants were able to restrict search to an appropriate subset of the display rather than conduct an exhaustive search. However, the young adults were more efficient in their ability to do so than were either the 7- or 10-year-old participants. The 10-year-olds were as efficient as the young adults when the number of black stimuli in the display was relatively small. However, these children became relatively less able to restrict search effectively as the number of black stimuli increased. Discussion focused on possible preattentive and attentive processes that may change systematically with age. PMID- 15336920 TI - The human ankle during walking: implications for design of biomimetic ankle prostheses. AB - The non-disabled human ankle joint was examined during walking in an attempt to determine overall system characteristics for use in the design of ankle prostheses. The hypothesis of the study was that the quasi-stiffness of the ankle changes when walking at different walking speeds. The hypothesis was examined using sagittal plane ankle moment versus ankle angle curves from 24 able-bodied subjects walking over a range of speeds. The slopes of the moment versus ankle angle curves (quasi-stiffness) during loading appeared to change as speed was increased and the relationship between the moment and angle during loading became increasingly non-linear. The loading and unloading portions of the moment versus angle curves showed clockwise loops (hysteresis) at self-selected slow speeds that reduced essentially to zero as the speed increased to self-selected normal speeds. Above self-selected normal speeds, the loops started to traverse a counter-clockwise path that increased in area as the speed was increased. These characteristics imply that the human ankle joint could be effectively replaced with a rotational spring and damper for slow to normal walking speeds. However, to mimic the characteristics of the human ankle during walking at fast speeds, an augmented system would be necessary. This notion is supported by the sign of the ankle power at the time of opposite heel contact, which was negative for slow speeds, was near zero at normal speeds, and was positive for fast walking speeds. PMID- 15336921 TI - Joint rotation between two attitudes in the spherical rotation coordinate system. AB - Three-dimensional joint rotation between two attitudes rather than starting from neutral position is the most common pattern of human movement. This paper presents a method for determining the joint rotations starting from a non-neutral attitude in the spherical rotation coordinate system based on a two-step rotation method. The method uses three spherical rotation angles that are consistent with clinical description of joint rotations and are mathematically proved to be sequence independent. The method can be easily understood by clinicians and applied in clinical practice. Two numerical examples of humeral rotations are presented to demonstrate the effectiveness of the method in dealing with different patterns of three-dimensional joint rotations between two attitudes. PMID- 15336922 TI - Adaptation to sudden unexpected loading of the low back--the effects of repeated trials. AB - The purpose of this study was to investigate short-term changes in reactions to sudden unexpected loading of the low back. The study utilized a set-up where a horizontal force of 58 N pointing forward suddenly was applied to the upper part of the subject's trunk. EMG activity from the erector spinae muscles and trunk movement data were recorded during 10 trials for 19 subjects. The analysis included EMG reaction time, mean rectified EMG amplitude during the period 50-250 ms after the sudden loading, and time elapsed until stopping of the forward movement of the trunk (stopping time). Reaction time means ranged from 66 to 97 ms (79+/-9 ms), and no difference was found between the trials. Conversely, the mean stopping time for the first trial (468 ms) was significantly higher than for trials 3-10 (359- 371 ms), and the average EMG amplitude during the period 50-250 ms after the sudden loading was lower for the first trial. This study showed that some subjects adapted to sudden unexpected loadings of the low back through a reduction in stopping time and a progression in EMG response during the first few trials. This possible adaptation to repeated trials have been overlooked in previous studies. PMID- 15336923 TI - Viscoelastic properties of skin in Mov-13 and Tsk mice. AB - Viscoelastic properties of skin samples were measured in three types of mice (tight skin, Tsk, control and Mov-13), that are known to differ with regard to content of type I collagen. The experimental design used uniaxial stretching and measured the creep response and the complex compliance. The creep response was measured directly. The complex compliance was determined using a Wiener-Volterra constitutive model for each sample. The models were calculated from data obtained by applying a stress input having a pseudo-Gaussian waveform and measuring the strain response. The storage compliance of Mov-13 and control skin were similar and were greater than Tsk (p<0.001). The loss compliance of each group was significantly different (p<0.001) from each other group; Tsk had the lowest and control had the highest loss compliance. The phase angle of the Mov-13 and Tsk were similar and were less than the controls (p<0.001). The creep response was fit with a linear viscoelastic model. None of the parameters in the creep model differed between groups. The results indicate that gene-targeted and mutant animals have soft tissue mechanical phenotypes that differ in complex ways. Caution should be exercised when using such animals as models to explore the role of specific constituents on tissue properties. PMID- 15336924 TI - In vivo tibiofemoral contact analysis using 3D MRI-based knee models. AB - This paper quantified the motion of the tibiofemoral contact points during in vivo weight bearing flexion using MRI- based 3D knee models and two orthogonal fluoroscopic images. The contact points on the medial and lateral tibial plateau were calculated by finding the centroid of the intersection of the tibial and femoral cartilage layers and by using the bony geometry alone. Our results indicate that the medial femoral condyle remains in the central portion of the tibial plateau and the lateral condyle translates posteriorly with increasing flexion. Using the bony contact model increased the total translation of the medial and lateral condyles by 250 and 55%, respectively, compared to the cartilage contact model. These results suggest that using the bony geometry alone may not accurately represent the articular surfaces of the knee. Articular cartilage geometry may have to be used to accurately quantify tibiofemoral contact. PMID- 15336925 TI - A fatigue damage model for the cement-bone interface. AB - Loss of fixation at the cement-bone interface can contribute to clinical loosening of cemented total hip replacements. In this study, the fatigue damage response was determined for cement-bone constructs subjected to shear fatigue loading. A typical three-phase fatigue response was observed with substantial early damage, followed by a long constant damage rate region and a final abrupt increase in damage to fracture. All of the damage resulted from creep (permanent) deformation during fatigue loading and there was no loss in cyclic stiffness. Using a Von Mises equivalent stress/strain concept, a general damage model was developed to describe the fatigue creep response of the cement-bone interface under either shear or tensile fatigue loading. Time to failure was highly correlated (r2=0.971) with equivalent creep strain rate and moderately related (r2=0.428) with equivalent initial strain for the two loading regimes. The equivalent creep strain at failure (0.052+/-0.018) was found to be independent of the applied equivalent stress. A combination of the creep damage model (to describe the damage process) with a constant final equivalent strain (as a failure criteria) could be used to assess the cement-bone failure response of cemented implant systems. PMID- 15336926 TI - Bending properties, porosity, and ash fraction of black bear (Ursus americanus) cortical bone are not compromised with aging despite annual periods of disuse. AB - In many species, including humans, disuse causes an imbalance in bone remodeling that leads to increased bone porosity as a result of increased bone resorption and decreased bone formation. However, black bears (Ursus americanus) may not develop disuse osteopenia, to the extent that other animals do, during long periods of disuse (i.e. hibernation) because they maintain osteoblastic bone formation during hibernation, even though bone resorption is increased during hibernation. Black bears may also have a mechanism to rapidly and completely recover the bone lost (by increased resorption during hibernation) during their remobilization period. Our findings suggest that cortical bone bending strength (211-328 MPa), bending modulus (16.0-29.5 MPa), fracture energy (0.0118-0.0205 J mm(-2)), porosity (2.3-7.1%), and ash fraction (0.638-0.672) are not compromised with age in black bears, despite annual periods of disuse. In fact, the ultimate strength (p=0.01), modulus (p=0.04), and ash fraction (p=0.03) of cortical bone were shown to significantly increase with age (2-14 yrs). Female bears give birth and nurse during hibernation; however, we found no significant (p>0.16) differences between male and female bone properties. Other animals require remobilization periods 2-3 times longer than the immobilization period to recover the bone lost during disuse. Our findings support the idea that black bears, which hibernate 5-7 months annually, have evolved a biological mechanism to mitigate the adverse effects of disuse on bone porosity and mechanical behavior. PMID- 15336927 TI - Design and validation of a bioreactor for engineering vocal fold tissues under combined tensile and vibrational stresses. AB - Criteria are outlined for the design of a bioreactor that can simulate the vibrational stresses in vocal fold movement during speech. Requirements are 0-1 mm amplitudes in the 20-200 Hz frequency range, a variable on-off stress regime, and maintenance of tissue viability over several days. The bioreactor uses dual drivers, one for low frequency (or static) strains, and another for high frequencies vibrational strains. Response is linear at the driving end for an input of 0-5 V. The amplitude decreases linearly with frequency at constant input voltage, and the phase changes by nearly 180 degrees over the 20-200 Hz range. Human vocal fold fibroblasts were cultured in a polymer substrate and subjected to static and vibrational forces. The results indicate that vibratory strain alters the expression levels of many extracellular matrix-related genes, as well as the spatial distribution of cells and matrix. PMID- 15336928 TI - Mechanical stimulation and mitogen-activated protein kinase signaling independently regulate osteogenic differentiation and mineralization by calcifying vascular cells. AB - Ectopic calcification of vascular tissue is associated with several cardiovascular pathologies and likely involves active regulation by vascular smooth muscle cells and osteoblast-like vascular cells. This process often occurs in sites with altered mechanical environments, suggesting a role for mechanical stimuli in calcification. In this study, we investigated the effect of mechanical stimulation on the proliferation, osteogenic differentiation, calcification, and mitogen-activated protein kinase (MAPK) signaling in calcifying vascular cells (CVCs), a subpopulation of aortic smooth muscle cells putatively involved in vascular calcification. Application of equibiaxial cyclic strain (7%, 0.25 Hz) to CVCs had no effect on cell proliferation, but accelerated alkaline phosphatase expression and significantly increased mineralization by 3.1-fold over unstrained cells. Fluid motion in the absence of strain also enhanced mineralization, but to a lesser degree. Because MAPK pathways mediate mechanically regulated osteoblast differentiation, we tested whether similar signaling was involved in mineralization by CVCs. In static cultures, pharmacological inhibition of the extracellular signal-regulated kinase (ERK1/2), p38 MAPK, and c-Jun N-terminal kinase pathways significantly attenuated mineral production by as much as -94%, compared with uninhibited CVCs. Strikingly, although mechanical stimulation activated each of the MAPK pathways, inhibition of these pathways had no effect on the mechanically induced enhancement of alkaline phosphatase activity or mineralization. These novel data indicate that mechanical signals regulate calcification by CVCs, and although MAPK signaling is critical to CVC osteogenic differentiation and mineralization, it is not involved directly in transduction of mechanical signals to regulate these processes under the conditions utilized in this study. PMID- 15336929 TI - Proliferation and collagen production of human patellar tendon fibroblasts in response to cyclic uniaxial stretching in serum-free conditions. AB - We studied the effect of cyclic mechanical stretching on the proliferation and collagen mRNA expression and protein production of human patellar tendon fibroblasts under serum-free conditions. The role of transforming growth factor beta1 (TGF-beta1) in collagen production by cyclically stretched tendon fibroblasts was also investigated. The tendon fibroblasts were grown in microgrooved silicone dishes, where the cells were highly elongated and aligned with the microgrooves. Cyclic uniaxial stretching with constant frequency and duration (0.5 Hz, 4 h) but varying magnitude of stretch (no stretch, 4%, and 8%) was applied to the silicone dishes. Following the period of stretching, the cells were rested for 20 h in stretching-conditioned medium to allow for cell proliferation. In separate experiments, the cells were stretched for 4h and then rested for another 4 h. Samples of the medium, total cellular RNA and protein were used for analysis of collagen and TGF-beta1 gene expression and production. It was found that there was a slight increase in fibroblast proliferation at 4% and 8% stretch, compared to that of non-stretched fibroblasts, where at 8% stretch the increase was significant. It was also found that the gene expression and protein production of collagen type I and TGF-beta1 increased in a stretching magnitude-dependent manner. And, levels of collagen type III were not changed, despite gene expression levels of the protein being slightly increased. Furthermore, the exogenous addition of anti-TGF-beta1 antibody eliminated the increase in collagen type I production under cyclic uniaxial stretching conditions. The results suggest that mechanical stretching can modulate proliferation of human tendon fibroblasts in the absence of serum and increase the cellular production of collagen type I, which is at least in part mediated by TGF-beta1. PMID- 15336930 TI - A new approach for surface fitting method of articular joint surfaces. AB - The application of joint contact mechanics requires a precise configuration of the joint surfaces. B-Spline, and NURBS have been widely used to model joint surfaces, but because these formulations use a structured data set provided by a rectangular net first, then a grid, there is a limit to the accuracy of the models they can produce. However new imaging systems such as 3D laser scanners can provide more realistic unstructured data sets. What is needed is a method to manipulate the unstructured data. We created a parametric polynomial function and applied it to unstructured data sets obtained by scanning joint surfaces. We applied our polynomial model to unstructured data sets from an artificial joint, and confirmed that our polynomial produced a smoother and more accurate model than the conventional B-spline method. Next, we applied it to a diarthrodial joint surface containing many ripples, and found that our function's noise filtering characteristics smoothed out existing ripples. Since no formulation was found to be optimal for all applications, we used two formulations to model surfaces with ripples. First, we used our polynomial to describe the global shape of the objective surface. Minute undulations were then specifically approximated with a Fourier series function. Finally, both approximated surfaces were superimposed to reproduce the original surface in a complete fashion. PMID- 15336931 TI - Testing the biomechanical optimality of the wall thickness of limb bones in the red fox (Vulpes vulpes). AB - The optimum for the ratio K of the internal to external diameter of a marrow filled tubular bone with minimum mass designed to withstand a given type of strength (yield/fatigue, stiffness, fracture or impact) depends on Q = rhom/rhob only, where rhom and rhob are the densities of marrow and bone. With computer assisted evaluation of radiographs of 62 femurs in the red fox (Vulpes vulpes) we measured the values of K. The mean and standard deviation of K are 0.68 and 0.036, and K changes in the rather wide range from 0.59 to 0.74. Accepting the assumption of earlier authors that Q = 0.50 or 0.44, our data would support the hypothesis that the fox femurs are optimized to withstand yield, fatigue or stiffness strengths. However, since the Q-values are unknown, the possibility cannot be excluded that any studied fox bone with an appropriately selected Q value is optimized for any strength type. Assuming Q = 0.50 or 0.44, the relative mass increments mu of the investigated fox bones are smaller than 5% under all four mechanical conditions. The evolutionary relevance of such tiny mu-values is questionable. PMID- 15336932 TI - Correlation between EMG and COP onset latency in response to a horizontal platform translation. AB - This study examined the relationship between onset latencies estimates from EMG and center of pressure (COP) in young (five female, five male; mean=24.2+/-2.3 years) and older (six female, four male; 78.4+/-2.3 years) subjects during anterior or posterior platform translations. The latencies to onset of activity were estimated for the tibialis anterior (TA; mean=119.8 ms across both age groups) and COP (mean=139.7 ms across both groups) for anterior translations, and the soleus (SOL; mean=122.4 ms across both groups), gastrocnemius (GAS; mean=126.0 ms for young, and 115.9 ms for old subjects) and COP (mean=160.0 ms across both groups) for posterior translations. Average within-subject correlations (r') among these measures showed a high correlation between TA and COP onset latency (r'=0.667, young; r'=0.482, old), and relatively low correlations between the plantar flexors (SOL and GAS) and COP onset latencies (SOL: r'=0.292 for young, r'=0.249 for old; GAS: r'=0.126 for young, r'=0.143 for old). The SOL and GAS onset latencies correlated well with each other, especially in the older subjects (r'=0.762), suggesting that the contribution of two muscles creates some variability in the relationship with COP onset latency. The strong correlation between TA and COP for anterior perturbations, coupled with the weaker correlations for the plantar flexors suggest that the COP method may be preferable for studies interested in determining timing of postural responses to multidirectional perturbations. PMID- 15336933 TI - Muscle activity in the leg is tuned in response to impact force characteristics. AB - Based on results from quasi-static experiments, it has been suggested that the lower extremity muscle activity is adjusted in reaction to impact forces with the goal of minimizing soft-tissue vibrations. It is not known whether a similar muscle tuning occurs during dynamic activities. Thus, the purpose of this study was to determine the effect of changes in the input signal on (a) vibrations of lower extremity soft-tissue packages and (b) EMG activity of related muscles during heel-toe running. Subjects performed heel-toe running in five different shoe conditions. Ground reaction forces were measured with a KISTLER force platform, soft-tissue vibrations were measured with tri-axial accelerometers and muscle activity was measured using surface EMG from the quadriceps, hamstrings, tibialis anterior and triceps surae groups from 10 subjects. By changing both the speed of running and the shoe midsole material the impact force characteristics were changed. There was no effect of changes in the input signal on the soft tissue peak acceleration following impact. A significant correlation (R2=0.819) between the EMG pre-activation intensity and the impact loading rate changes was found for the quadriceps. In addition, the input frequency was shown to approach the vibration frequency of the quadriceps. This evidence supports the proposed paradigm that muscle activity is tuned to impact force characteristics to control the soft-tissue vibrations. PMID- 15336934 TI - Finger joint impedance during tapping on a computer keyswitch. AB - We studied the dynamic behavior of finger joints during the contact period of tapping on a computer keyswitch, to characterize and parameterize joint function with a lumped-parameter impedance model. We tested the hypothesis that the metacarpophalangeal (MCP) and interphalangeal (IP) joints act similarly in terms of kinematics, torque, and energy production when tapping. Fifteen human subjects tapped with the index finger of the right hand on a computer keyswitch mounted on a two-axis force sensor, which measured forces in the vertical and sagittal planes. Miniature fiber-optic goniometers mounted across the dorsal side of each joint measured joint kinematics. Joint torques were calculated from endpoint forces and joint kinematics using an inverse dynamic algorithm. For each joint, a linear spring and damper model was fitted to joint torque, position, and velocity during the contact period of each tap (22 per subject on average). The spring damper model could account for over 90% of the variance in torque when loading and unloading portions of the contact were separated, with model parameters comparable to those previously measured during isometric loading of the finger. The finger joints functioned differently, as illustrated by energy production during the contact period. During the loading phase of contact the MCP joint flexed and produced energy, whereas the proximal and distal IP joints extended and absorbed energy. These results suggest that the MCP joint does work on the interphalangeal joints as well as on the keyswitch. PMID- 15336935 TI - Automatic generation of accurate subject-specific bone finite element models to be used in clinical studies. AB - Most of the finite element models of bones used in orthopaedic biomechanics research are based on generic anatomies. However, in many cases it would be useful to generate from CT data a separate finite element model for each subject of a study group. In a recent study a hexahedral mesh generator based on a grid projection algorithm was found very effective in terms of accuracy and automation. However, so far the use of this method has been documented only on data collected in vitro and only for long bones. The present study was aimed at verifying if this method represents a procedure for the generation of finite element models of human bones from data collected in vivo, robust, accurate, automatic and general enough to be used in clinical studies. Robustness, automation and numerical accuracy of the proposed method were assessed on five femoral CT data sets of patients affected by various pathologies. The generality of the method was verified by processing a femur, an ileum, a phalanx, a proximal femur reconstruction, and the micro-CT of a small sample of spongy bone. The method was found robust enough to cope with the variability of the five femurs, producing meshes with a numerical accuracy and a computational weight comparable to those found in vitro. Even when the method was used to process the other bones the levels of mesh conditioning remained within acceptable limits. Thus, it may be concluded that the method presents a generality sufficient to cope with almost any orthopaedic application. PMID- 15336936 TI - A new acceleration apparatus for the study of whiplash with human cadaveric cervical spine specimens. AB - The biomechanics of whiplash is often studied using cadaveric cervical spine specimens. One of the most important points in this kind of study is to create realistic loading conditions. The aim of the present project therefore was to develop an acceleration apparatus, which allows the study of whiplash with human cadaveric cervical spine specimens under as realistic loading conditions as possible. The new acceleration apparatus mainly consisted of a sled, a pneumatic acceleration unit and a railtrack and offered several unique features to create more realistic loading conditions. Among these features, the possibility to simulate the passive movements of the trunk is of capital importance. In this new apparatus, first, the general feasibility of whiplash experiments was studied, second, the reproducibility of the impacts was quantified and third, the effect of simulated movements of the trunk on accelerations and loads was examined. In the new acceleration apparatus various types of collisions could reproducibly be simulated. Simulated passive movements of the trunk strongly influenced the loading pattern of the neck. Without pivoting a steep increase of all loading parameters could be observed. This increase was less pronounced if pivoting was allowed. In conclusion, biomechanical aspects of whiplash could reproducibly be examined in the new acceleration apparatus. Due to its significant effects on the loading of the neck, pivoting of the trunk should always be taken into account in future experiments on the biomechanics of whiplash in which isolated cervical spine specimens are used. PMID- 15336937 TI - An ultrasound-based method for determining pulse wave velocity in superficial arteries. AB - In this paper, we present a method for estimating local pulse wave velocity (PWV) solely from ultrasound measurements: the area-flow (QA) method. With the QA method, PWV is estimated as the ratio between change in flow and change in cross sectional area (PWV = dQ/dA) during the reflection-free period of the cardiac cycle. In four anaesthetized dogs and 21 human subjects (age 23-74) we measured the carotid flow and cross-sectional area non-invasively by ultrasound. As a reference method we used the Bramwell-Hill (BH) equation which estimates PWV from pulse pressure and cross-sectional area. Additionally, we therefore measured brachial pulse pressure by oscillometry in the human subjects, and central aortic pulse pressure by micro-manometry in the dogs. As predicted by the pressure dependency of arterial stiffness, the estimated PWV decreased when the aortic pressure was lowered in two of the dogs. For the human subjects, the QA and BH estimates were correlated (R=0.43, p<0.05) and agreed on average (mean difference of -0.14 m/s). The PWV by the BH method increased with age (p<0.01) whereas the PWV by the QA method tended to increase with age (p<0.1). This corresponded to a larger residual variance (residual = deviation of the estimated PWV from the regression line) for the QA method than for the BH method, indicating different precisions for the two methods. This study illustrates that the simple equation PWV = dQ/dA gives estimates correlated to the PWV of the reference method. However, improvements in the basic measurements seem necessary to increase the precision of the method. PMID- 15336938 TI - Three-dimensional measurements of the pressure distribution in artificial joints with a capacitive sensor array. AB - A spherically folded capacitive pressure sensor array is introduced and characterized. By placing the sensor array between the ball and the cavity of artificial joints, the pressure distribution within the joint was recorded with spatial resolution for different size matching between the ball and the cavity, for different directions of loading and for joints with incomplete cavities. The performance of the sensor array is analyzed, possible fields of application as well as its limitations are discussed. PMID- 15336939 TI - Obesity and knee joint torque and power during level walking. PMID- 15336941 TI - alpha,beta-MeATP augments the UTP contraction of rabbit basilar artery. Yasushi Miyagi, Nitoshi Kimura, Robin C. Carpenter, Andrew D. Parent, John Zhang [Eur. J. Pharmacol. 488 (2004) 117-125]. PMID- 15336942 TI - Altered response to mirtazapine on gene expression profile of lymphocytes from Alzheimer's patients. AB - Antidepressants are widely used in the treatment of mood disorders associated with dementia, however little information is available on their effect at the molecular level. We have demonstrated that gene expression profiles of lymphocytes from patients with Alzheimer dementia differ from that seen with controls, with alpha(2)-adrenoceptor being the most highly repressed transcript. To address this issue in light of antidepressant treatment, we used lymphocytes derived from Alzheimer patients and control individuals to assess the impact of mirtazapine, the novel antidepressant with alpha(2)-adrenoceptor antagonistic activities, on gene expression using a cDNA microarray representing 3200 distinct human genes. Sequences that are differentially regulated after treatment with mirtazapine were identified and categorized based on similarities in biological functions. This analysis revealed that selected biological processes, including protein metabolism, cytoskeleton integrity, immune response, cellular plasticity, and neurotransmission, are involved in early phases of administration of this antidepressant. In addition, although it was possible to identify common targets, the expression profiles of Alzheimer lymphocytes differed mainly in their magnitude from those seen with controls. These results confirm the usefulness of the gene array approach for studying Alzheimer-specific changes in the periphery and suggest that the expression of genes of Alzheimer lymphocytes is modulated differently by mirtazapine, which correlates with the pathology. PMID- 15336943 TI - Effects of interleukin-1beta, interleukin-13 and transforming growth factor-beta on gene expression in human airway smooth muscle using gene microarrays. AB - Inflammatory gene expression in airway smooth muscle may be influenced by its inflammatory milieu. We analysed the gene expression profile of airway smooth muscle cells cultured from human airways exposed to a pro-inflammatory cytokine, interleukin-1beta, a T helper-2 cytokine, interleukin-13, and to a growth factor, transforming growth factor (TGF)beta1 (10 ng/ml each) after 4 and 24 h using the Affymetrix GeneChip 95A array which detects approximately 12,500 genes and expression sequence tags (ESTs). Airway smooth muscle cells were responsive to each cytokine with distinctive patterns of gene expression for cytokines, chemokines, adhesion and signalling proteins, and transcription factors. Interleukin-1beta induced the highest number of genes such as cytokines/chemokines including interleukin-8, growth-related oncogene (GRO) alpha, -beta and -gamma, epithelial neutrophil activating protein (ENA)-78, monocyte chemotactic protein (MCP)-1, -2 and -3 and eotaxin. Using quantitative real-time reverse transcription-polymerase chain reaction, the expression of GRO alpha, -beta and -gamma, interleukin-8 and eotaxin by interleukin-1beta was confirmed, with good correlation with microarray data. Transforming growth factor (TGF)beta1 induced other growth factors such as connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF), insulin growth factor (IGF) and many structural and extracellular matrix proteins. Interleukin-13 was the weakest inducer, with stimulation of eotaxin and genes of unknown function. While many genes were co-regulated at 4 and 24 h, there were also differences in expression patterns. Interleukin-1beta induces a predominantly pro-inflammatory profile while TGFbeta1 can be linked to proliferative and matrix changes. The rich profile of mediators, growth factors and signalling molecules released from airway smooth muscle depends on the inflammatory milieu. PMID- 15336944 TI - Anti-apoptotic action of (2S,3S,4R)-N"-cyano-N-(6-amino-3,4-dihydro-3-hydroxy-2 methyl-2-dimethoxymethyl-2H-benzopyran-4-yl)-N'-benzylguanidine (KR-31378) by suppression of the phosphatase and tensin homolog deleted from chromosome 10 phosphorylation and increased phosphorylation of casein kinase2/Akt/ cyclic AMP response element binding protein via maxi-K channel opening in neuronal cells. AB - This study shows the signaling pathway by which (2S,3S,4R)-N"-cyano-N-(6-amino 3,4-dihydro-3-hydroxy-2-methyl-2-dimethoxymethyl-2H-benzopyran-4-yl)-N' benzylguanidine (KR-31378) prevents tumor necrosis factor (TNF)-alpha-induced neuronal cell death. KR-31378 restored TNF-alpha-induced decreased cell viability of SK-N-SH. U87-MG cells (PTEN-null glioblastoma cell line) transfected with expression vectors for sense PTEN (phosphatase and tensin homolog deleted from chromosome 10) showed significantly decreased cell viability, which was restored by KR-31378. TNF-alpha-induced increased PTEN phosphorylation and decreased phosphorylation of Akt/cyclic AMP response element-binding protein (CREB) in SK-N SH cells were concentration-dependently reversed by KR-31378, those of which were antagonized by iberiotoxin, a maxi-K channel blocker. TNF-alpha and apigenin, a casein kinase2 (CK2) inhibitor, showed decreased CK2 phosphorylation and increased PTEN phosphorylation, which were reversed by KR-31378. KR-31378 increased K(+) currents by activating the maxi-K channels in SK-N-SH cells, with suppression of TNF-alpha-induced increase in cytosolic Ca(2+) and elevation of suppressed mitochondrial membrane potential, all of which were antagonized by iberiotoxin. It is suggested that increase in cell viability by KR-31378 is ascribed to the maxi-K channel opening-coupled upregulation of CK2/Akt/CREB phosphorylation and downregulation of PTEN phosphorylation in association with increased Bcl-2 and decreased Bax levels. PMID- 15336945 TI - Effects of (-)-linalool in the acute hyperalgesia induced by carrageenan, L glutamate and prostaglandin E2. AB - A series of studies performed in our laboratory have shown that (-)-linalool, the natural occurring enantiomer in essential oils, possesses anti-inflammatory and antinociceptive effects in different animal models. The antinociceptive effect of (-)-linalool has been ascribed to the stimulation of the cholinergic, opioidergic and dopaminergic systems, to its local anesthetic activity and to the blockade of N-methyl-D-aspartate (NMDA) receptors. In this study, we investigated the effect of systemic administration of (-)-linalool in the paw withdrawal test in rats, a model of thermal hyperalgesia induced by monolateral subplantar injection of carrageenan, L-glutamate or prostaglandin E(2). Carrageenan and L-glutamate induced a hyperalgesic effect on the injection side. In contrast, prostaglandin E(2) induced hyperalgesia in both the injection side and the contralateral side. Pretreatment with (-)-linalool (50-150 mg/kg) inhibited the development of acute hyperalgesia induced by carrageenan in the injected paw, with no effect on the contralateral paw. Furthermore, (-)-linalool at the highest dose used (200 mg/kg), reduced and reverted the decrease in paw withdrawal latencies induced by L-glutamate on the ipsilateral side, showing antihyperalgesic and antinociceptive effects. An antinociceptive effect was apparent also in the contralateral paw. Finally, (-)-linalool (200 mg/kg) increased paw withdrawal latency on the side contralateral to prostaglandin E(2) injection, but not on the side of the injection. The efficacy of (-)-linalool in decreasing the hyperalgesia induced by carrageenan, L-glutamate and prostaglandin E(2) suggests that this compound might be useful in pain conditions sustained by the development of neuronal sensitization. PMID- 15336946 TI - Fluoxetine attenuates thermal hyperalgesia through 5-HT1/2 receptors in streptozotocin-induced diabetic mice. AB - Diabetic neuropathic pain, an important microvascular complication in diabetes mellitus, is recognised as one of the most difficult types of pain to treat. A lack of understanding of its aetiology, inadequate relief, development of tolerance and potential toxicity of classical antinociceptives warrant the investigation of newer agents to relieve this pain. The aim of the present study was to explore the antinociceptive effect and possible mechanism of action of a serotonin reuptake inhibitor, fluoxetine, in streptozotocin-induced diabetic mice. Four weeks after a single intraperitoneal injection of streptozotocin (200 mg/kg), mice were tested in the tail-immersion and hot-plate assays. Diabetic mice exhibited significant hyperalgesia compared with control mice. Fluoxetine (10 and 20, but not 5 mg/kg, i.p.) injected into diabetic mice produced an antinociceptive effect in both the tail-immersion and hot-plate assays. The percentage maximum possible effect (% MPE) produced by fluoxetine (20 mg/kg, i.p.) was significantly lower in diabetic mice than in control mice. The antinociceptive effect of fluoxetine (20 mg/kg) in diabetic mice was dose dependently potentiated by pindolol (5 and 10 mg/kg, i.p., a selective 5 HT(1A/1B) receptor antagonist), attenuated by ritanserin (1 and 2 mg/kg, i.p., a selective 5-HT(2A/2C) receptor antagonist) and remained unaffected by ondansetron (1 and 2 mg/kg, i.p., a selective 5-HT(3) receptor antagonist) in both test systems. These results suggest that fluoxetine-induced antinociception primarily involves serotonin pathway modulation through 5-HT(1) and 5-HT(2) receptors, but not through 5-HT(3) receptors, in the chronic pain associated with streptozotocin induced diabetic neuropathy. Further, the potentiation of the antinociceptive effect of fluoxetine by pindolol indicates the usefulness of a combination of an antidepressant and a 5-HT(1A/1B) receptor antagonist in the treatment of diabetic neuropathic pain in humans. PMID- 15336947 TI - Sensory nitrergic meningeal vasodilatation and non-nitrergic plasma extravasation in anaesthesized rats. AB - The aim of the present study was to evaluate the role of nitric oxide (NO) of sensory neural origin in neurogenic inflammatory response in the trigeminovascular system. Antidromic vasodilatation and plasma extravasation in response to electrical stimulation (15 V, 5 Hz, 0.5 ms, 100 impulses) of the trigeminal ganglion were investigated in the dura mater and nasal mucosa/upper eyelid by laser Doppler flowmetry and [(125)I]-labelled bovine serum albumin, respectively. Electrical stimulation of the trigeminal ganglion of rats elicited a reproducible ipsilateral enhancement of both meningeal and nasal mucosal blood flow. N(omega)-nitro-L-arginine (L-NNA; 4, 8, and 16 mg/kg, i.v.), a nonselective inhibitor of nitric oxide synthase (NOS), inhibited antidromic vasodilatation both in the dura mater (15.86+/-2.05%, 22.82+/-2.51%, and 36.28+/-4.37%) and nasal mucosa (35.46+/-8.57%, 58.72+/-9.2%, and 89.99+/-8.94%) in a dose-dependent manner. Specific inhibitors of neuronal NOS, 7-nitroindazole (7-NI; 20 mg/kg, i.v.) and 3-bromo-7-nitroindazole (3Br-7NI; 10 mg/kg, i.v.) were administered to assess the possible role of NO released from the trigeminal sensory fibres. The meningeal vasodilatation was inhibited by both 3Br-7NI and 7-NI (63.36+/-7.7% and 49+/-6.5%, respectively). The nasal hyperaemic response was also reduced by 3Br 7NI (78.26+/-8.7%). Plasma extravasation in the dura mater and upper eyelid evoked by electrical stimulation of the trigeminal ganglion (25 V, 5 Hz, 0,5 ms, 5 min), expressed as extravasation ratios (ERs) of the stimulated vs. nonstimulated sides, was 1.80+/-0.8 and 4.63+/-1.24, respectively. This neurogenic oedema formation was not inhibited by neither L-NNA nor 3Br-7NI. It is concluded that neural nitrergic mechanisms are involved in the meningeal vasodilatation evoked by electrical stimulation of the trigeminal ganglion. PMID- 15336948 TI - [Ethyl-3H]RS-79948-197 alpha2-adrenoceptor autoradiography validation in alpha2 adrenoceptor knockout mice. AB - [Ethyl-(3)H][8aR,12aS,13aS]-5,8,8a,9,10,11,12,12a,13,13a-decahydro-3-methoxy-12 (ethylsulfonyl)-6H-isoquino[2,1-g][1,6]naphthyridine ([ethyl-(3)H]RS-79948-197) was evaluated for alpha(2)-adrenoceptor autoradiography in brain sections from wild-type mice and alpha(2A)- and alpha(2ABC)-adrenoceptor knockout mice. Receptor numbers were 83% lower in cortex and 28% lower in caudate putamen of alpha(2A)-knockout mice than in wild-type mice. No specific binding was seen in alpha(2ABC)-knockout mice. [Ethyl-(3)H]RS-79948-197 saturation binding parameters were compared to those of [(3)H]2-(2,3-dihydro-2-methoxy-1,4-benzodioxan-2-yl) 4,5-dihydro-1H-imidazoline ([(3)H]RX821002) and [methyl-(3)H]17alpha-hydroxy 20alpha-yohimban-16beta-carboxylic acid methyl ester ([methyl-(3)H]rauwolscine). [Ethyl-(3)H-]RS-79948-197 detected a larger number of both alpha(2A)- and alpha(2B/C)-adrenoceptors than [(3)H]RX821002, while [methyl-(3)H]rauwolscine only underestimated the number of alpha(2A)-adrenoceptors. Oxymetazoline and prazosin competed for [ethyl-(3)H]RS-79948-197 binding with the expected rank order of affinities. Higher than necessary [ethyl-(3)H]RS-79948-197 concentrations resulted in a rapid increase in non-specific binding. Slow dissociation kinetics, high specific radioactivity and high alpha(2)-adrenoceptor affinity (slightly lower for the alpha(2A)-adrenoceptor than for the other subtypes) confer [ethyl-(3)H]RS-79948-197 distinct advantages compared to [(3)H]RX821002 for detection of alpha(2)-adrenoceptor subtypes in a mixed alpha(2)-adrenoceptor population. PMID- 15336949 TI - Selective serotonin reuptake inhibitor reduces conditioned fear through its effect in the amygdala. AB - Selective serotonin reuptake inhibitors are first-line treatment for most anxiety disorders, but their mechanism of anxiolytic action has not been clarified. Selective serotonin reuptake inhibitors are anxiolytic in conditioned fear stress (re-exposure to an environment paired previously with inescapable electric footshocks). To clarify the brain regions where selective serotonin reuptake inhibitors act, we examined the effect of microinjection of the selective serotonin reuptake inhibitor, citalopram, into the amygdala, medial prefrontal cortex and mediodorsal nucleus of the thalamus on freezing behavior, an index of fear, induced by conditioned fear stress. Bilateral injection of citalopram into the amygdala before testing reduced freezing significantly, while bilateral injection into the medial prefrontal cortex or mediodorsal nucleus of the thalamus did not. These results suggest that the anxiolytic effect of a selective serotonin reuptake inhibitor in conditioned fear is mediated by its effect in the amygdala, and support the hypothesis of serotonin function in anxiety by which facilitation of serotonin neurotransmission decreases anxiety. PMID- 15336950 TI - Characterization of endothelium-dependent relaxation and modulation by treatment with pioglitazone in the hypercholesterolemic rabbit renal artery. AB - The present study was undertaken to investigate vascular function in hypercholesterolemic rabbits and also to characterize the effects of pioglitazone on it. Rabbits were fed normal, 0.5% cholesterol chow, or 0.5% cholesterol chow plus 300 ppm pioglitazone for 5 or 10 weeks. The tension of isolated renal artery rings was measured isometrically, and morphometric analysis was performed. The cholesterol chow diet administered for 5 weeks did not affect acetylcholine induced relaxation in the renal artery but that for 10 weeks decreased it. The N(G)-nitro-L-arginine (L-NOARG)- and indomethacin-resistant endothelium-dependent relaxation induced by acetylcholine in the renal artery was enhanced in rabbits receiving the cholesterol chow for 5 or 10 weeks, as compared to rabbits receiving the control diet, and the percentage of plaque area formation was increased in the renal artery by the cholesterol chow for 10 weeks. Pioglitazone normalized them without lowering serum lipid levels. The resistant parts of acetylcholine-induced relaxation was significantly inhibited when the renal artery was treated with charybdotoxin, an inhibitor of large and intermediate conductance Ca(2+)-activated K(+) channels, or N,N-diethylaminoethyl-2,2 diphenylvalerate hydrochloride (SKF 525a), a cytochrome P-450 monooxygenase inhibitor. Results indicate that hypercholesterolemia enhances endothelium derived hyperpolarizing factor (EDHF)-mediated relaxation in the rabbit renal artery and pioglitazon normalizes it without lowering serum lipid levels, and suggest that the maintenance of endothelial function by pioglitazon is related to the mechanisms for its anti-atheromatous activity. PMID- 15336951 TI - Effects of tamsulosin on hypogastric nerve stimulation-induced intraurethral pressure elevation in male and female dogs under anesthesia. AB - The aim of the present study was to investigate the effects of tamsulosin, an alpha(1)-adrenoceptor antagonist, on hypogastric nerve stimulation-induced intraurethral pressure elevation in anesthetized male and female dogs and to evaluate sex differences in these effects. Additionally, the effects of tamsulosin were also compared with those of other alpha(1)-adrenoceptor antagonists, namely prazosin, naftopidil and urapidil. Tamsulosin dose dependently inhibited hypogastric nerve stimulation-induced intraurethral pressure elevation, with doses required to induce 50% inhibition of the elevation (ED(50) values) of 0.72 and 0.74 microg/kg i.v. in anesthetized male and female dogs, respectively. Mean arterial blood pressure slightly decreased after administration of tamsulosin at a dose which inhibited intraurethral pressure elevation almost completely. Prazosin, naftopidil and urapidil also inhibited increases in intraurethral pressure in a dose-dependent fashion, but caused decreases in mean arterial blood pressure at the same doses. The estimated rank order of inhibitory potency for urethral response was tamsulosin>prazosin>naftopidil=urapidil. In conclusion, tamsulosin dose dependently inhibited increases in intraurethral pressure with little effect on mean arterial blood pressure in both male and female dogs, and these effects were almost equipotent. These results indicate that tamsulosin will be useful in the treatment of dysuria associated with lower urinary tract symptoms in women as well as men. PMID- 15336952 TI - Vitamin A reduces lung granulomatous inflammation with eosinophilic and neutrophilic infiltration in Sephadex-treated rats. AB - Vitamin A is known to suppress the activity of the transcription factors, nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1), as do glucocorticoids. The possibility that vitamin A exerts various anti-inflammatory effects therefore seems likely. Sephadex beads were administered intravenously to anesthesized rats pretreated with a subcutaneous injection of vitamin A (3000, 10,000, or 30,000 IU/kg) or vehicle once daily for 3 days. After 16 h, the leukocyte differential, tumor necrosis factor (TNF)-alpha and eotaxin, and the DNA-binding activity of NF kappaB were measured in bronchoalveolar lavage fluid (BALF). Additionally, lung histology was assessed using preparations stained with May-Giemsa stain. Sephadex beads caused histological granulomatous changes and eosinophilic and neutrophilic infiltration into the lung, and markedly increased cell counts of eosinophils and neutrophils, concentrations of TNF-alpha and eotaxin, and NF-kappaB binding to DNA in BALF. Vitamin A significantly inhibited all responses. Vitamin A may inhibit Sephadex-induced lung granulomatous formation, and eosinophilic and neutrophilic infiltration due to its suppression of TNF-alpha and eotaxin production, and NF-kappaB activation. PMID- 15336953 TI - The role of CCL3/macrophage inflammatory protein-1alpha in experimental colitis. AB - CCL3/macrophage inflammatory protein (MIP-1)-1alpha is elevated in the rectal biopsies of patients with active inflammatory bowel diseases, but its role remains undefined. The present study examined the role of CCL3/MIP-1alpha during trinitrobenzene sulfonic acid (TNBS)-induced colitis in the rat. Colonic CCL3/MIP 1alpha levels were elevated (>20-fold above control) within 24 h and remained elevated to day 7 of colitis induction by TNBS administration. In addition, significant increases in colonic neutrophil accumulation were observed within 24 h to day 7 of TNBS treatment. Pre-treatment of rats with a single dose of CCL3/MIP-1alpha antibody significantly reduced (47%) colonic neutrophil accumulation during the early (24 h) phase of TNBS-induced colitis. In contrast, chronic (repeated) administration of CCL3/MIP-1alpha antibody did not attenuate colonic neutrophil accumulation during the late phase (day 7) of TNBS-induced colitis. These results suggest a role for CCL3/MIP-1alpha in promoting colonic neutrophil accumulation during the early (24 h) phase of TNBS-induced colitis. PMID- 15336954 TI - Combination benefit of a pyrimidylpiperazine derivative (Y-40138) and methotrexate in arthritic rats. AB - Anti-tumor necrosis factor-alpha (TNFalpha) antibody in combination with methotrexate dramatically decreases joint destruction in rheumatoid arthritis. The aim of this study was to examine combined treatment with N-[1-(4-([4 (pyrimidin-2-yl)piperazin-1-yl]methyl)phenyl)cyclopropyl] acetamide HCl (Y-40138) and methotrexate in rat adjuvant-induced arthritis. The increase in hindpaw volume and joint destruction was suppressed by single therapeutic administration (days 15-20) of Y-40138 (30 mg/kg, p.o.), but not by prophylactic administration (days 1-9). However, arthritic progression was suppressed by single prophylactic administration of methotrexate (0.3 mg/kg, p.o.), but not by therapeutic administration. Combined administration (days 10-20) of Y-40138 (0.3-1 mg/kg) and methotrexate (0.03 mg/kg) synergistically suppressed the increase in hindpaw volume and joint destruction. We concluded that Y-40138 in combination with methotrexate synergistically suppressed arthritic progression. These data suggest that combined treatment with Y-40138 and methotrexate may increase efficacy of therapy for rheumatoid arthritis. PMID- 15336956 TI - Calpain-related diseases. AB - Calpains are calcium-modulated proteases which respond to Ca2+ signals by removing limited portions of protein substrates, thereby irreversibly modifying their function(s). Members of this protease family are present in a variety of organisms ranging from mammals to plants; some of them are ubiquitously expressed, while others are tissue specific. Although calpains are apparently involved in a multitude of physiological and pathological events, their functions are still poorly understood. In two cases, however, the alteration of a member of the calpain family has been clearly identified as being responsible for a human disease: the loss of function of calpain 3 causes limb girdle muscular dystrophy type 2A, and mutations in the gene coding for calpain 10 have been shown to correlate with non-insulin-dependent diabetes. PMID- 15336957 TI - Ca2+ binding protects against gelsolin amyloidosis. AB - Amyloid diseases occur when native or mutant polypeptides misfold and aggregate to form deposits in the extracellular space. There are at least 20 proteins associated with amyloid diseases, including the well-known amyloid-beta peptide that is the causative agent for Alzheimer's disease (AD). This review describes familial amyloidosis of Finnish type (FAF), an amyloid disease caused by mutations in plasma gelsolin, a secreted protein that contains multiple Ca2+ binding domains. The FAF mutations result in a loss of the Ca2+-binding site in domain 2 of plasma gelsolin. The resulting decreased stability gives rise to susceptibility to the protease furin in the Golgi. Furin cleavage generates a secreted fragment that undergoes a second proteolytic event in the extracellular matrix to produce a peptide that self-assembles into amyloid plaques. Thus, Ca2+ binding in native plasma gelsolin protects against amyloid disease. PMID- 15336958 TI - S100 proteins in mouse and man: from evolution to function and pathology (including an update of the nomenclature). AB - The S100 protein family is the largest subgroup within the superfamily of proteins carrying the Ca2+-binding EF-hand motif. Despite their small molecular size and their conserved functional domain of two distinct EF-hands, S100 proteins developed a plethora of tissue-specific intra- and extracellular functions. Accordingly, various diseases such as cardiomyopathies, neurodegenerative and inflammatory disorders, and cancer are associated with altered S100 protein levels. Here, we review the different S100 protein functions and related diseases from an evolutionary point of view. We analyzed the structural variations, which are the basis of functional diversification, as well as the genomic organization of the S100 family in human and compared it with the S100 repertoires in mouse and rat. S100 genes and proteins are highly conserved between the different mammalian species. Moreover, we identified evolutionary related subgroups of S100 proteins within the three species, which share functional similarity and form subclusters on the genomic level. The available S100-specific mouse models are summarized and the consequences of our results are discussed with regard to the use of genetically engineered mice as human disease models. An update of the S100 nomenclature is included, because some of the recently identified S100 genes and pseudogenes had to be renamed. PMID- 15336959 TI - Guanylate cyclase-activating proteins: structure, function, and diversity. AB - The guanylate cyclase-activating proteins (GCAPs), Ca2+-binding proteins of the calmodulin gene superfamily, function as regulators of photoreceptor guanylate cyclases. In contrast to calmodulin, which is active in the Ca2+-bound form, GCAPs stimulate GCs in the [Ca2+]-free form and inhibit GCs upon Ca2+ binding. In vertebrate retinas, at least two GCAP1 and two GCs are present, a third GCAP3 is expressed in humans and fish, and at least five additional GCAP4-8 genes have been identified or are predicted in zebrafish and pufferfish. Missense mutations in GCAP1 (Y99C, I143NT, E155G, and P50L) have been associated with autosomal dominant cone dystrophy. Absence of GCAP1/2 in mice delays recovery of the photoresponse, a phenotype consistent with delay in cGMP synthesis. In the absence of GCAP2, GCAP1 supports the generation of wild-type flash responses in both rod and cone cells. Recent progress revealed an unexpected complexity of the GC-GCAP system, pointing, out a number of unsolved questions. PMID- 15336960 TI - Hippocalcin in the olfactory epithelium: a mediator of second messenger signaling. AB - Intracellular Ca2+ plays an important role in a variety of second messenger cascades. The function of Ca2+ is mediated, in part, by Ca2+-binding proteins such as calmodulin, calretinin, calbindin, neurocalcin, recoverin, and visinin like proteins (VILIPs). These proteins are highly expressed in rat olfactory receptor neurons (ORNs) and are localized to distinct intracellular regions. In the present study, we have identified another Ca2+-binding protein, hippocalcin, in the rat olfactory epithelium (OE). Olfactory/brain hippocalcin shows high sequence homology with hippocalcins expressed in mice and humans. Hippocalcin was predominantly localized to the olfactory cilia, the site of the initial events of olfactory signal transduction, and was found to regulate the activity of ciliary adenylate cyclases (ACs) and particulate guanylyl cyclases (GCs) in a Ca2+ dependent manner. These data indicate that hippocalcin is expressed in rat ORNs, and is likely to regulate second messenger cascades in a Ca2+-dependent manner. PMID- 15336961 TI - A role for calsenilin and related proteins in multiple aspects of neuronal function. AB - The protein variously called calsenilin, DREAM, or KChIP3 has been independently discovered three times, accounting for the three different names currently in use. Calsenilin appears to have three, perhaps independent, roles. Calsenilin binds and modulates some of the effects of the Alzheimer disease-related protein presenilin, while presenilin can modulate some of the effects of calsenilin. Calsenilin binds the dynorphin response element and regulates dynorphin expression, hence regulating nociception. Calsenilin binds Kv channels and modulates potassium conductance, playing a role in long-term potentiation as well as in other important plastic pathways. Other members of the calsenilin family share at least some of the roles. For example, KChIP1, KChIP2, and CALP can all bind presenilins and can all modulate A-type potassium channels. Further functional dissection of this family of proteins will provide insight into numerous aspects of neuronal function and will illuminate the role of the calsenilin family of proteins in disease. PMID- 15336962 TI - Presenilin regulates capacitative calcium entry dependently and independently of gamma-secretase activity. AB - Mutations in presenilin-1 and 2 (PS) lead to increased intracellular calcium stores and an attenuation in the refilling mechanism known as capacitative calcium entry (CCE). Previous studies have shown that the mechanism by which PS modulates intracellular calcium signaling is dependent on gamma-secretase activity. Although the modulation of intracellular calcium signaling can lead to alterations in CCE, it is plausible that PS can also directly affect CCE independent of the effect it exerts on intracellular stores. To investigate this possibility, we studied the effects of the dominant negative variant of PS1 known as DeltaTM1-2, which lacks the first two transmembrane domains of PS1 and in which gamma-secretase activity is abrogated. We demonstrate that, like other dominant negative isoforms of PS1, DeltaTM1-2 expression leads to reduced intracellular calcium. However, unlike other dominant negative isoforms, DeltaTM1 2 leads to a deficit rather than a potentiation of CCE. These data suggest that changes in the structural components of presenilin can modulate CCE independent of its function in gamma-secretase activity and intracellular calcium stores. PMID- 15336963 TI - Cancer-related diseases of the eye: the role of calcium and calcium-binding proteins. AB - The eye provides unique opportunities to study complex biochemical pathways and to describe how components of these pathways contribute to the molecular basis of disease. In this article, the role of calcium-binding proteins in cancer-related diseases of the eye is reviewed. First, paraneoplastic syndromes, or so-called remote effects of cancer, arise from damage to tissues distant from any tumor or its metastases. Many of these syndromes are believed to be immune-mediated. Cancer-associated retinopathy (CAR), a blinding disease due to the degeneration of retinal photoreceptor cells, is one of the best characterized of the paraneoplastic syndromes. The CAR autoantigen has been identified as recoverin, a calcium-binding protein of the EF-hand superfamily. Its features as a calcium binding protein, along with its function in photoreceptor cells and its role as the CAR autoantigen, are discussed. Next, unlike visual symptoms instigated by a distant tumor, ocular melanoma is the primary malignancy originating in the eye. ALG-2 encodes a pro-apoptotic calcium-binding protein that is down-regulated in ocular melanoma, thus providing these tumor cells with a selective advantage. In addition to background discussion of ALG-2, data describing the expression, cellular localization, and dimerization characteristics of ALG-2 in melanoma cells are presented. Biochemical studies of ALG-2 and its interactions with its target Alix/AIP1 also are presented. Finally, the function of ALG-2 in calcium induced cell death is discussed. Additional calcium-binding proteins in retina and in ocular tumors are described in relation to different disease entities. Such proteins and their expression in the eye provide valuable examples bridging studies of protein chemistry, cellular function, and human disease. PMID- 15336964 TI - Annexins and disease. AB - The annexins are a family of closely related calcium- and membrane-binding proteins expressed in most eukaryotic cell types. Despite their structural and biochemical similarities annexins have diverse functions, in cellular activities that include vesicle trafficking, cell division, apoptosis, calcium signalling, and growth regulation. To date there is no evidence to suggest that any individual member of the annexin family is a disease-causing gene, i.e., a gene that through loss, mutation, translocation or amplification leads to a known human disease. However, there is good evidence that in certain clinical conditions, changes in annexin expression levels or localisation may contribute to the pathological consequences and sequelae of disease. In this way, annexins are indirectly linked to some of the most serious human disease classes including cardiovascular disease and cancer. In this review we consider the roles played by annexins in disease and examine the molecular basis for anomalous annexin behaviour that may contribute to disease pathophysiology. PMID- 15336965 TI - Annexin VI regulation of cardiac function. AB - Annexins are a family of membrane binding proteins that are characterized by a hypervariable amino terminus followed by a series of highly conserved Ca2+ phospholipid binding domains. Annexins function by binding to anionic phospholipid surfaces in a Ca2+-dependent manner. They self-associate to form trimers which further assemble into sheets that cover the membrane surface and alter properties such as fluidity and permeability. This submembranous skeleton alters integral protein functions such as ion transport properties and shields the surface from phospholipid binding proteins such as phospholipases and protein kinase C. Transgenic mouse hearts overexpressing wild type annexin VI (AnxVI673), a dominant-negative truncated annexin VI (residues 1-129, Anx129) and an annexin VI-null mouse (AnxVI-/-) have implicated the protein as a regulator of intracellular Ca2+ homeostasis which affects cardiac function. PMID- 15336966 TI - Calcium-calcineurin signaling in the regulation of cardiac hypertrophy. AB - Cardiac hypertrophy is a leading predicator of progressive heart disease that often leads to heart failure and a loss of cardiac contractile performance associated with profound alterations in intracellular calcium handling. Recent investigation has centered on identifying the molecular signaling pathways that regulate cardiac myocyte hypertrophy, as well as the mechanisms whereby alterations in calcium handling are associated with progressive heart failure. One potential focal regulator of cardiomyocyte hypertrophy that also responds to altered calcium handling is the calmodulin-activated serine/threonine protein phosphatase calcineurin (PP2B). Once activated by increases in calcium, calcineurin mediates the hypertrophic response through its downstream transcriptional effector nuclear factor of activated T cells (NFAT), which is directly dephosphorylated by calcineurin resulting in nuclear translocation. While previous studies have convincingly demonstrated the sufficiency of calcineurin to mediate cardiac hypertrophy and progressive heart failure, its necessity remains an area of ongoing investigation. Here we weigh an increasing body of literature that suggests a causal link between calcineurin signaling and the cardiac hypertrophic response and heart failure through the use of pharmacologic inhibitors (cyclosporine A and FK506) and genetic approaches. We will also discuss the manner in which calcineurin-NFAT signaling is negatively regulated in the heart through a diverse array of kinases and inhibitory proteins. Finally, we will discuss emerging theories as to the mechanisms whereby alterations in intracellular calcium handling might stimulate calcineurin within the context of a contractile cell continually experiencing calcium flux. PMID- 15336967 TI - Phenotypes of SERCA and PMCA knockout mice. AB - P-type Ca2+-ATPases of the sarco(endo)plasmic reticulum (SERCAs) and plasma membrane (PMCAs) are responsible for maintaining the Ca2+ gradients across cellular membranes that are required for regulation of Ca2+-mediated signaling and other biological processes. Gene-targeting studies of SERCA isoforms 1, 2, and 3 and PMCA isoforms 1, 2, and 4 have confirmed some of the general functions proposed for these pumps, such as a major role in excitation-contraction coupling for SERCA1 and SERCA2 and housekeeping functions for PMCA1 and SERCA2, but have also revealed some unexpected phenotypes. These include squamous cell cancer and plasticity in the regulation of Ca2+-mediated exocytosis in SERCA2 heterozygous mutant mice, modulation of Ca2+ signaling in SERCA3-deficient mice, deafness and balance disorders in PMCA2 null mice, and male infertility in PMCA4 null mice. These unique phenotypes provide new information about the cellular functions of these pumps, the requirement of their activities for higher order physiological processes, and the pathophysiological consequences of pump dysfunction. PMID- 15336968 TI - SPCA1 pumps and Hailey-Hailey disease. AB - Both the endoplasmic reticulum and the Golgi apparatus are agonist-sensitive intracellular Ca2+ stores. The Golgi apparatus has Ca2+-release channels and a Ca2+-uptake mechanism consisting of sarco(endo)plasmic-reticulum Ca2+-ATPases (SERCA) and secretory-pathway Ca2+-ATPases (SPCA). SPCA1 has been shown to transport both Ca2+ and Mn2+ in the Golgi lumen and therefore plays an important role in the cytosolic and intra-Golgi Ca2+ and Mn2+ homeostasis. Human genetic studies have provided new information on the physiological role of SPCA1. Loss of one functional copy of the SPCA1 (ATP2C1) gene causes Hailey-Hailey disease, a skin disorder arising in the adult age with recurrent vesicles and erosions in the flexural areas. Here, we review recent experimental evidence showing that the Golgi apparatus plays a much more important role in intracellular ion homeostasis than previously anticipated. PMID- 15336969 TI - Sarcoplasmic reticulum Ca-ATPase-phospholamban interactions and dilated cardiomyopathy. AB - Dilated cardiomyopathy is a disease of the heart muscle resulting from a diverse array of conditions that damages the heart and impairs myocardial function. Heart failure occurs when the heart is unable to pump blood at a rate which can accommodate the heart muscle's metabolic requirements. Several signaling pathways have been shown to be involved in the induction of cardiac disease and heart failure. Many of these pathways are linked to cardiac sarcoplasmic reticulum (SR) Ca cycling directly or indirectly. A large body of evidence points to the central role of abnormal Ca handling by SR proteins, Ca-ATPase pump (SERCA2a) and phospholamban (PLN), in pathophysiological heart conditions, compromising the contractile state of the cardiomyocytes. This review summarizes studies which highlight the key role of these two SR proteins in the regulation of cardiac function, the significance of SERCA2a-PLN interactions using transgenic approaches, and the recent discoveries of human PLN mutations leading to disease states. Finally, we will discuss extrapolation of experimental paradigms generated in animal models to the human condition. PMID- 15336970 TI - Endoplasmic reticulum calcium transport ATPase expression during differentiation of colon cancer and leukaemia cells. AB - The calcium homeostasis of the endoplasmic reticulum (ER) is connected to a multitude of cell functions involved in intracellular signal transduction, control of proliferation, programmed cell death, or the synthesis of mature proteins. Calcium is accumulated in the ER by various biochemically distinct sarco/endoplasmic reticulum calcium transport ATPase isoenzymes (SERCA isoforms). Experimental data indicate that the SERCA composition of some carcinoma and leukaemia cell types undergoes significant changes during differentiation, and that this is accompanied by modifications of SERCA-dependent calcium accumulation in the ER. Because ER calcium homeostasis can also influence cell differentiation, we propose that the modulation of the expression of various SERCA isoforms, and in particular, the induction of the expression of SERCA3-type proteins, is an integral part of the differentiation program of some cancer and leukaemia cell types. The SERCA content of the ER may constitute a new parameter by which the calcium homeostatic characteristics of the organelle are adjusted. The cross-talk between ER calcium homeostasis and cell differentiation may have some implications for the better understanding of the signalling defects involved in the acquisition and maintenance of the malignant phenotype. PMID- 15336971 TI - Darier's disease: from dyskeratosis to endoplasmic reticulum calcium ATPase deficiency. AB - The skin is the body's largest organ and has an essential barrier protective function against physical, chemical, and pathogen aggressions and prevents fluid loss. The outer layer of the skin, known as the epidermis, plays a key role in this protection, through a tightly regulated differentiation programme from basal keratinocytes to the stratum corneum at the skin surface. During this process, keratinocytes from the base of the epidermis undergo major morphological and functional changes during their migration through the spinous and granular layers, to become terminally differentiated corneocytes which will be shed from the skin's surface. The role of extracellular Ca2+ in cell-to-cell adhesion and in epidermal differentiation was known to be important, but the identification of the sarco/endoplasmic reticulum Ca2+ transport ATPase (ATP2A2) as the defective gene in a rare genetic skin disease known as Darier's disease, came as a surprise and shed light on the key role of Ca2+ signaling in the homeostasis of the epidermis. PMID- 15336972 TI - Ryanodine receptor defects in muscle genetic diseases. AB - Ryanodine receptor (RyR), a homotetrameric Ca2+ release channel, is one of the main actors in the generation of Ca2+ signals that trigger muscle contraction. Three genes encode three isoforms of RyRs, which have tissue-restricted distribution. RyR1 and RyR2 are typical of muscle cells, with RyR1 originally considered the skeletal muscle type and RyR2 the cardiac type. However, RyR1 and RyR2 have recently been found in numerous other cell types, including, for instance, peripheral B and T lymphocytes. In contrast, RyR3 is widely distributed among cells. RyR1 and RyR2 are localized in a specialized portion of the sarcoplasmic reticulum (SR), the terminal cisternae, which is the portion of the SR Ca2+ store that releases Ca2+ to control the process of muscle contraction. A specific role for RyR3 has not yet been established: probably, its co-expression with the other RyR isoforms contributes to qualitatively modulate Ca2+-dependent processes in muscle cells and in neurons. Several mutations in the genes encoding RyR1 and RyR2 have been identified in autosomal dominant diseases of skeletal and cardiac muscle, such as malignant hyperthermia (MH), central core disease (CCD), catecholaminergic polymorphic ventricular tachycardia (CPVT), and arrhythmogenic right ventricular dysplasia type 2 (ARVD2). More recently, CCD cases with recessive inheritance have also been described. MH is a pharmacogenetic disease, but the others manifest as congenital myopathies. Even if their clinical phenotypes are well established, particularly in skeletal muscle, the molecular mechanisms that generate the conditions are not clear. A number of studies on cellular models have attempted to elucidate the molecular defects associated with the different mutations, but the problem of understanding how mutations in the same gene generate such an array of diverse pathological traits and diseases of widely different degrees of severity is still open. This review will consider the molecular and cellular effects of RyR mutations, summarizing recent data in the literature on Ca2+ dysregulation, which may lead to a better understanding of the functioning of RyRs. PMID- 15336973 TI - Dynamic alterations in myoplasmic Ca2+ in malignant hyperthermia and central core disease. AB - Ca2+ ions play a pivotal role in a wide array of cellular processes ranging from fertilization to cell death. In skeletal muscle, a mechanical interaction between plasma membrane dihydropyridine receptors (DHPRs, L-type Ca2+ channels) and Ca2+ release channels (ryanodine receptors, RyR1s) of the sarcoplasmic reticulum orchestrates a complex, bi-directional Ca2+ signaling process that converts electrical impulses in the sarcolemma into myoplasmic Ca2+ transients during excitation-contraction coupling. Mutations in the genes that encode the two proteins that coordinate this electrochemical conversion process (the DHPR and RyR1) result in a variety of skeletal muscle disorders including malignant hyperthermia (MH), central core disease (CCD), multiminicore disease, nemaline rod myopathy, and hypokalemic periodic paralysis. Although RyR1 and DHPR disease mutations are thought to alter excitability and Ca2+ homeostasis in skeletal muscle, only recently has research begun to probe the molecular mechanisms by which these genetic defects lead to distinct clinical and histopathological manifestations. This review focuses on recent advances in determining the impact of MH and CCD mutations in RyR1 on muscle Ca2+ signaling and how these effects contribute to disease-specific aspects of these disorders. PMID- 15336974 TI - Calstabin deficiency, ryanodine receptors, and sudden cardiac death. AB - Altered cardiac ryanodine receptor (RyR2) function has an important role in heart failure and genetic forms of arrhythmias. RyR2 constitutes the major intracellular Ca2+ release channel in the cardiac sarcoplasmic reticulum (SR). The peptidyl-prolyl isomerase calstabin2 (FKBP12.6) is a component of the RyR2 macromolecular signaling complex. Calstabin2 binding to RyR2 is regulated by PKA phosphorylation of Ser2809 in RyR2. PKA phosphorylation of RyR2 decreases the binding affinity for calstabin2 and increases RyR2 open probability and sensitivity to Ca2+-dependent activation. In heart failure, a majority of studies have found that RyR2 becomes chronically PKA hyper-phosphorylated which depletes calstabin2 from the channel complex. Calstabin2 dissociation causes a diastolic SR Ca2+ leak contributing to depressed intracellular Ca2+ cycling and decreased cardiac contractility. Missense mutations linked to genetic forms of exercise induced arrhythmias and sudden cardiac death also cause decreased calstabin2 binding affinity and leaky RyR2 channels. We review the importance of calstabin2 for RyR2 function and excitation-contraction coupling, and discuss new observations that implicate dysregulation of calstabin2 binding as a central mechanism for abnormal calcium cycling in heart failure and triggered arrhythmias. PMID- 15336975 TI - Ryanodine receptor channelopathies. AB - Ryanodine receptors (RyR) are the Ca2+ release channels of sarcoplasmic reticulum that provide the majority of the [Ca2+] necessary to induce contraction of cardiac and skeletal muscle cells. In their cellular environment, RyRs are exquisitely regulated by a variety of cytosolic factors and accessory proteins so that their output signal (Ca2+) induces cell contraction without igniting signaling pathways that eventually lead to contractile dysfunction or pathological cellular remodeling. Here we review how dysfunction of RyRs, most commonly expressed as enhanced Ca2+ release at rest (skeletal muscle) or during diastole (cardiac muscle), appears to be the fundamental mechanism underlying several genetic or acquired syndromes. In skeletal muscle, malignant hyperthermia and central core disease result from point mutations in RYR1, the skeletal isoform of RyRs. In cardiac muscle, RYR2 mutations lead to catecholaminergic polymorphic ventricular tachycardia and other cardiac arrhythmias. Lastly, an altered phosphorylation of the RyR2 protein may be involved in some forms of congestive heart failure. PMID- 15336976 TI - Myocardial calcium signalling and arrhythmia pathogenesis. AB - Myocardial calcium signalling is a vital component of the normal physiological function of the heart. Key amongst the many roles calcium plays is its use as the primary signalling component of excitation-contraction coupling, the intracellular process that links cardiomyocyte depolarisation to contraction. Defective cellular calcium handling, due to abnormalities of the various components which mediate and control excitation-contraction coupling, is widely recognised as a significant patho-physiological event in the contractile dysfunction of the failing heart. In addition, similar defects also appear to be increasingly recognised as mediators of certain forms of cardiac arrhythmias. Such defects include single gene defects in excitation-contraction coupling components that lead to inherited sudden death arrhythmia syndromes. Alternatively, arrhythmogenesis occurring within the context of acquired cardiac disease, in particular heart failure, also appears to be highly dependent on abnormal calcium homeostasis. In this article we review the defects in cardiomyocyte calcium homeostasis that lead to particular pro-arrhythmogenic phenomena and discuss recent insights gained into a variety of inherited and acquired arrhythmia syndromes that appear to involve defective calcium signalling as a central component of their patho-physiology. Potential opportunities for new anti arrhythmic therapeutic strategies based on these recent insights are also discussed. PMID- 15336977 TI - Deranged neuronal calcium signaling and Huntington disease. AB - Huntington disease (HD) is an autosomal-dominant neurodegenerative disorder that primarily affects medium spiny striatal neurons (MSN). HD is caused by polyglutamine (polyQ) expansion (exp) in the amino-terminal region of a protein huntingtin (Htt). The connection between polyQ expansion in Httexp and MSN neurodegeneration remains elusive. Here we discuss recent data that link polyQ expansion in Httexp and deranged Ca2+ signaling in MSN neurons. Experimental evidence indicates that (1) Ca2+ homeostasis is abnormal in mitochondria isolated from lymphoblasts of HD patients and from brains of the YAC72 HD mouse model; (2) Httexp leads to potentiation of NR1/NR2B NMDA receptor activity in heterologous expression systems and in MSN from YAC72 HD mouse model; and (3) Httexp binds to the type 1 inositol 1,4,5-trisphosphate receptor (InsP3R1) carboxy-terminus and causes sensitization of InsP3R1 to activation by InsP3 in planar lipid bilayers and in MSN. Based on these results we propose that Httexp-induced cytosolic and mitochondrial Ca2+ overload of MSN plays an important role in the pathogenesis of HD and that Ca2+ signaling blockers may play a beneficial role in treatment of HD. PMID- 15336978 TI - The role of inositol 1,4,5-trisphosphate receptors in the regulation of bile secretion in health and disease. AB - Ca2+ signaling via the inositol 1,4,5-trisphosphate receptor (InsP3R) is a ubiquitous mechanism for regulation of cell function, yet very little is known about the role of the InsP3R in specific disease states. Converging lines of evidence suggest that the liver may provide a model for the role of the InsP3R in health and disease. Ca2+ signaling is mediated entirely by the InsP3R in hepatocytes and cholangiocytes, the two types of epithelia in the liver. Here we review the role of specific InsP3R isoforms and the physiological effects of InsP3R-mediated Ca2+ signals in both of these types of epithelia. In addition, we review evidence that the InsP3R is lost from cholangiocytes in cholestatic forms of liver disease, and discuss this as a possible final common pathway for cholestasis. PMID- 15336979 TI - Ca2+ homeostasis in apoptotic resistance of prostate cancer cells. AB - Ca2+ is a universal messenger regulating many physiological functions including such an important one, as the ability of the cell to undergo orderly self destruction upon completion of its mission, called apoptosis. If this function is compromised unwanted cells may eventually take over the tissue turning it into a cancer. Ca2+ dependency of apoptosis, when its all aspects are learned and understood and key molecular players identified, may provide a good opportunity for controlling tumor growth. In the present mini-review we describe the major molecular determinants of Ca2+ homeostasis in prostate cancer cells and establish their role in the transformation to apoptosis-resistant cell phenotypes typical of advanced androgen-independent prostate cancer. We show that the hallmark of such transformation is the inhibition of apoptosis pathway associated with endoplasmic reticulum Ca2+ store depletion. PMID- 15336980 TI - Genetic manipulation of cardiac Na+/Ca2+ exchange expression. AB - The Na+/Ca2+ exchanger (NCX) is the primary Ca2+ extrusion mechanism in cardiomyocytes. To further investigate the role of NCX in excitation-contraction coupling and Ca2+ homeostasis, we created murine models with altered expression levels of NCX. Homozygous overexpression of NCX resulted in mild cardiac hypertrophy. Decline of the Ca2+ transient and relaxation of contraction were increased and the reverse mode of NCX was augmented. Overexpression also led to a higher susceptibility to ischemia-reperfusion injury and to a greater ability of NCX to trigger Ca2+-induced Ca2+ release. Furthermore, an increase in peak L-type Ca2+ current was observed suggesting a direct influence of NCX on L-type Ca2+ current. Whereas global knockout of NCX led to prenatal death, a recently generated cardiac-specific NCX knockout mouse was viable with surprisingly normal contractile properties. Expression levels of other Ca2+-handling proteins were not altered. Ca2+ influx in these animals is limited by a decrease of peak L-type Ca2+ current. An alternative Ca2+ efflux mechanism, presumably the plasma membrane Ca2+-ATPase, is sufficient to maintain Ca2+-homeostasis in the NCX knockout mice. PMID- 15336981 TI - L-type Ca2+ channels in Ca2+ channelopathies. AB - Voltage-gated L-type Ca2+ channels (LTCCs) mediate depolarization-induced Ca2+ entry in electrically excitable cells, including muscle cells, neurons, and endocrine and sensory cells. In this review we summarize the role of LTCCs for human diseases caused by genetic Ca2+ channel defects (channelopathies). LTCC dysfunction can result from structural aberrations within pore-forming alpha1 subunits causing incomplete congenital stationary night blindness, malignant hyperthermia sensitivity or hypokalemic periodic paralysis. However, studies in mice revealed that LTCC dysfunction also contributes to neurological symptoms in Ca2+ channelopathies affecting non-LTCCs, such as Ca(v)2.1 alpha1 in tottering mice. Ca2+ channelopathies provide exciting molecular tools to elucidate the contribution of different LTCC isoforms to human diseases. PMID- 15336982 TI - Lambert-Eaton myasthenic syndrome as an autoimmune calcium channelopathy. AB - Lambert-Eaton myasthenic syndrome, often associated with small-cell lung carcinoma, is a disease of neuromuscular transmission in which antibodies directed against voltage-gated calcium channel (VGCC)(P/Q-type) in the motor nerve terminal play a crucial role in causing a deficient quantal release of acetylcholine. The motor nerve terminal and carcinoma cell may share a common antigen. The study using synthetic peptides and recombinant protein specified the extracellular S5-S6 linker regions in 3 of 4 domains as immunodominant sites in the molecular structure of P/Q-type VGCC alpha1 subunit. Also, the study by use of peptides and recombinant protein corresponding to synaptotagmin I suggested that in this functionally VGCC-associated presynaptic protein, the segment which exposes extracellularly during exocytosis can be immunogenic for the syndrome. PMID- 15336983 TI - Functional role of TRPC proteins in vivo: lessons from TRPC-deficient mouse models. AB - In order to elucidate the functional role of TRPC genes, in vivo, the targeted inactivation of these genes in mice is an invaluable technique. In this review, we summarize the currently available results on the phenotype of TRPC-deficient mouse lines. The analysis of mice with targeted deletion in three TRPC genes demonstrates that these proteins represent essential constituents of agonist activated and phospholipase C-dependent Ca2+ entry channels in primary cells. Furthermore, from the deficits observed in these TRPC-deficient mouse lines a striking number of biological functions could already be ascribed to TRPC2, TRPC4, and TRPC6, not only on the cellular level but also for complex organ functions and integrative physiology. Accordingly, TRPC2 proteins are critically involved in pheromone sensing by neurones of the vomeronasal organ and, thereby, in the regulation of sexual and social behavior of mice, TRPC4 proteins are essential determinants of endothelial-dependent regulation of vascular tone, endothelial permeability, and neurotransmitter release from thalamic interneurones, and TRPC6 proteins are supposed to have a fundamental role in the regulation of smooth muscle tone in blood vessels and lung. PMID- 15336984 TI - TRPV6 and prostate cancer: cancer growth beyond the prostate correlates with increased TRPV6 Ca2+ channel expression. AB - Life expectancy for patients suffering from prostate cancer is inversely correlated with the degree of extraprostatic metastasis. In order to find pharmacological tools to treat this aggressive growth it is important to define targets whose expression not only correlates with the malignancy of the cancerous cells, but that are also amenable to pharmacological intervention. In this review, we would like to focus on the potential role of a distinct class of ion channels that may be involved in this process. PMID- 15336985 TI - Calcium signaling and polycystin-2. AB - Polycystic kidney disease (PKD) is caused by mutations in two genes, PKD1 and PKD2, which encode for the proteins, polycystin-1 (PC1) and polycystin-2 (PC2), respectively. Although disease-associated mutations have been identified in these two proteins, the sequence of molecular events leading up to clinical symptoms is still unknown. PC1 resides in the plasma membrane and it is thought to function in cell-cell and cell-matrix interactions, whereas PC2 is a calcium (Ca2+) permeable cation channel concentrated in the endoplasmic reticulum. Both proteins localize to the primary cilia where they function as a mechanosensitive receptor complex allowing the entry of Ca2+ into the cell. The downstream signaling pathway involves activation of intracellular Ca2+ release channels, especially the ryanodine receptor (RyR), but subsequent steps are still to be identified. Elucidation of the signaling pathway involved in normal PC1/PC2 function, the functional consequences of PC1/PC2 mutation, and the role of Ca2+ signaling will all help to unravel the molecular mechanisms of cystogenesis in PKD. PMID- 15336986 TI - Polycystins, calcium signaling, and human diseases. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a major, inherited nephropathy affecting over 1:1000 of the worldwide population. It is a systemic condition with frequent hepatic and cardiovascular manifestations in addition to the progressive development of fluid-filled cysts from the tubules and collecting ducts of affected kidneys. The pathogenesis of cyst formation is currently thought to involve increased proliferation of epithelial cells, mild dedifferentiation, and fluid accumulation. In the past decade, study of ADPKD led to the discovery of a unique family of highly complex proteins, the polycystins. Loss-of-function mutations in either of two polycystin proteins, polycystin-1 or polycystin-2, give rise to ADPKD. These proteins are thought to function together as part of a multiprotein complex that may initiate Ca2+ signals, directing attention to the regulation of intracellular Ca2+ as a possible misstep that participates in cyst formation. Here we review what is known about the Ca2+ signaling functions of polycystin proteins and focus on findings that have significantly advanced our physiological insight. Special attention is paid to the recently discovered role of these proteins in the mechanotransduction of the renal primary cilium and the model it suggests. PMID- 15336987 TI - Functional links between mucolipin-1 and Ca2+-dependent membrane trafficking in mucolipidosis IV. AB - Most of the membrane trafficking phenomena including those involving the interactions between endosomes and lysosomes are regulated by changes in intracellular Ca2+ (Cai). These processes are disturbed in some types of mucolipidoses and other lysosomal storage disorders, such as mucolipidosis IV (MLIV), a neurological disorder that usually presents during the first year of life with blindness, cognitive impairment, and psychomotor delays. It is caused by mutations in MCOLN1, the gene encoding mucolipin-1 (MLN1), which we have recently established to represent a Ca2+-permeable cation channel that is transiently modulated by changes in Cai. The cells of MLIV patients contain enlarged lysosomes that are likely associated with abnormal sorting and trafficking of these and related organelles. We studied fibroblasts from MLIV patients and found disturbed Ca2+ signaling and large acidic organelles such as late endosomes and lysosomes (LEL) with altered cellular localization in these cells. The fusion between LEL vesicles in these cells was defective. This is a Ca2+-dependent process related to signaling pathways involved in regulation of Ca2+ homeostasis and trafficking. The MLN1 channels could play a key role in Ca2+ release from LEL vesicles, which triggers the fusion and trafficking of these organelles. The characterization of this MLN1-mediated Ca2+-dependent process should provide new insights into the pathophysiological mechanisms that lead to the development of MLIV and other mucolipidoses associated with similar disturbances in membrane trafficking. PMID- 15336988 TI - A one-year comparison of prophylactic donor tricuspid annuloplasty in heart transplantation. AB - BACKGROUND: The bicaval technique for orthotopic heart transplantation decreases the incidence of tricuspid valve regurgitation when compared with the standard biatrial technique. This study was designed to study the effects of prophylactic tricuspid valve annuloplasty during bicaval orthotopic heart transplantation on survival, renal function, and amount of tricuspid valve regurgitation. METHODS: Between April 1997 and March 1998, 60 patients (age 18 to 70 years, 22 women) randomly received either bicaval orthotopic heart transplantation (n = 30) or bicaval orthotopic heart transplantation with DeVega tricuspid valve annuloplasty (n = 30). Tricuspid valve annuloplasty was performed on the donor heart before implantation using pledgeted 2-0 polypropylene suture and sized to an annulus of 29 mm. Echocardiographic variables, laboratory values, and hemodynamics were obtained prospectively and reviewed by an independent data analyst. RESULTS: Intraoperatively, the group undergoing tricuspid valve annuloplasty had a shorter reperfusion time (46 +/- 29 minutes versus 65 +/- 48 minutes; p < 0.05) and higher mean pulmonary artery to central venous pressure difference (11.8 +/- 3.7 mm Hg versus 15.3 +/- 4.1 mm Hg; p = 0.001). Additional differences between the two groups included early mortality from donor dysfunction (4 of 30 patients versus 0 of 30 patients; p < 0.05), amount of tricuspid valve regurgitation at 1 year (1.3 +/- 1.0 versus 0.2 +/- 0.3; p < 0.05), and percentage of patients with 2+ or greater tricuspid valve regurgitation (34% versus 0%; p < 0.05). CONCLUSIONS: Tricuspid valve annuloplasty of the donor heart before bicaval orthotopic heart transplantation improves immediate donor heart function as demonstrated by better right ventricular performance, lower perioperative mortality, and shorter reperfusion times. At 1 year, there is less tricuspid valve regurgitation but no difference in renal function. Considering the ease and safety of tricuspid valve annuloplasty and its advantages, it should be performed as a routine adjunct with bicaval orthotopic heart transplantation. PMID- 15336989 TI - Results of aortic valve-sparing operations: experience with remodeling and reimplantation procedures in 65 patients. AB - BACKGROUND: Valve-sparing operations for aortic root aneurysms are increasing in frequency, but techniques and results are still in evolution. We reviewed our experience with 65 patients (adults and children) who had this operation at our institution to determine early and late outcomes. METHODS: A retrospective clinical review was undertaken using hospital records, clinical and echocardiographic, computed tomography, magnetic resonance imaging data, and telephone interviews with patients and their physicians. RESULTS: Between July 1994 and December 2002, 65 patients (46 adults and 19 children) underwent a valve sparing operation for aortic root aneurysm. Forty-four of the patients had the Marfan syndrome; the remaining 21 had either a nonspecific connective tissue disorder (14 patients) or a miscellaneous disease process such as Ehlers-Danlos syndrome (7 patients). Fifty-eight (89%) had a David II (remodeling) procedure and 7 had a David I (reimplantation) procedure. The DePaulis "Valsalva graft" was used in six of the David I patients. There were no operative or hospital deaths; only one late death occurred in an adult due to salmonella meningitis. Overall, survival was 100% at one year and 98% at 3 and 5 years. Ten patients (7 adults and 3 children) developed significant late aortic insufficiency (AI). Nine of these patients had a David II procedure and in 8 of these cases, AI was secondary to significant late annular dilatation. One of the 10 patients developed late AI 8.2 years after a David I procedure; his AI was secondary to aortic leaflet extension and prolapse. Six of the 10 patients who developed significant late AI required aortic valve replacement (4 adults and 2 children). Freedom from late aortic valve replacement (AVR) in this series of 65 patients was 91% at 3 and 84% at 5 years. At the close of this study, 58 patients were New York Heart Association (NYHA) class I and 6 were NYHA class II; no patients were class III or IV. There were no episodes of endocarditis or clinically significant thromboembolism. CONCLUSIONS: Valve-sparing operations provide satisfactory results for many patients with an aortic root aneurysm, but the David II remodeling procedure has a greater risk of late annular dilatation and AI. The David I reimplantation procedure utilizing the DePaulis Valsalva graft may obviate this problem. PMID- 15336990 TI - The Ross procedure: long-term clinical and echocardiographic follow-up. AB - BACKGROUND: Progressive dilatation of the pulmonary autograft is the principal cause for reoperation following the Ross procedure when the root replacement technique is used. We examined the relation between enlargement of the pulmonary autograft and the development and progression of neo-aortic valve regurgitation, and the long-term clinical follow-up, including the need for reoperation, in patients followed for up to 13 years postoperatively. METHODS: A Ross procedure was performed on 119 older children and young adults (mean age: 31 years old), using the root replacement technique, between June 1989 and January 2002. Serial echocardiography studies were obtained in 108 patients and analyzed blinded to clinical data. The following variables were measured: diameter of annulus, sinuses of Valsalva, and supravalvular ridge; presence and severity of aortic regurgitation; and valve thickening. RESULTS: The 30 day and late mortality rates were 1.7% and 1.7% (2 patients each). Forty-one patients were followed for more than 5 years, 19 for more than 7 years, and 9 for more than 10 years. There was one thrombotic and no endocarditis events. The 10-year actuarial survival was 96%. Reoperation on the pulmonary autograft or the pulmonary allograft was required in 12 patients. The principal indication for operation on the pulmonary autograft in 11 patients was neo-aortic valve regurgitation (7), aneurysm formation (3), and false aneurysm (1). At 10 years, actuarial freedom from reoperation on the pulmonary autograft was 75%. At last follow-up, 8 of 97 patients without reoperation on the autograft had moderate and none had severe regurgitation of the neo-aortic valve. Independent predictors of progression of neo-aortic valve regurgitation were time from operation, dilatation of the supravalvular ridge, and neo-aortic valve thickening (all p < 0.0002). Freedom from reoperation in the pulmonary allograft at 10 years was 86%. CONCLUSIONS: Long-term follow-up of patients with the Ross procedure using the root replacement technique indicates excellent survival and low thromboembolic and endocarditis risk. The main limitation is the need for reoperation. The prevalence of severe neo-aortic valve regurgitation is low, however there is a progressive increase in regurgitation and in aortic root diameters. Periodic follow-up with echocardiography is recommended because of the continuing risk of progressive regurgitation of the neo-aortic valve and aneurysm formation. PMID- 15336991 TI - Redo-aortic valve replacement after previous bilateral internal thoracic artery bypass grafting. AB - BACKGROUND: Aortic valve replacement (AVR) after coronary artery bypass using bilateral internal thoracic arteries (ITAs) is a challenge. Management of these patent grafts and myocardial protection are important issues. Moreover the risk and outcome of these complex operations have not been clearly defined. METHODS: Eighteen consecutive patients (all male) who exhibited previous bilateral ITA grafts underwent subsequent AVR surgery from 1990-2001 at the Cleveland Clinic Foundation. Their medical records were retrospectively analyzed. RESULTS: At the time of reoperation, the mean age of the patients was 67 +/- 6.4 years and 33 out of 36 (92%) ITAs were patent. The interval between previous coronary bypass and aortic valve surgery was 10.3 +/- 5.3 years. All patients underwent redo-median sternotomy with aortic cannulation in 12 patients (67%) and femoral or axillary artery cannulation in 6 patients (33%). The patent ITAs were clamped during aortic cross-clamping in 15 patients. In 3 patients the ITAs were not dissected. These 3 patients underwent deep hypothermic arrest for myocardial protection. Concomitant coronary revascularization was performed in 8 patients (44%). There were no hospital deaths. One stroke occurred but there were no other major complications. Average intubation time was 23.1 +/- 27.1 hours, intensive care unit stay was 2.3 +/- 3.1 days, and postoperative hospital stay was 10.3 +/- 7.6 days. CONCLUSIONS: Reoperative aortic valve surgery in the patients with patent bilateral ITA grafts can be performed safely. PMID- 15336992 TI - St. Jude Medical Toronto biologic aortic root prosthesis: early FDA phase II IDE study results. AB - BACKGROUND: Several biological aortic root replacement techniques have distinct advantages over mechanical composite root replacement including better valvular hemodynamic characteristics and the lack of need for anticoagulation. Current biological root replacement options lack proven long-term durability or are limited by technical or practical concerns. We report the early results from a phase II multicenter clinical trial of the porcine St. Jude Toronto Bioprosthesis with BiLinx (Toronto root). METHODS: 176 Toronto roots were implanted as total aortic root replacement from August 2001 through August 2003. Concomitant cardiac procedures including coronary artery bypass grafting (31%) and ascending aortic replacement (55%) were performed in 74%. Patients were followed clinically and were examined with an echocardiogram at discharge, 6 months, 12 months, and yearly thereafter. Root sizes implanted included 29 mm in 38%, 27 mm in 30%, 25 mm in 20%, 23 mm in 10%, and 21 mm in 2.2%. RESULTS: There are 205 patient years of follow-up through October 2003. Operative mortality was 3.9% (none were valve related) and late mortality was 4%. Operative stroke rate was 1.1% and late stroke rate was 0.6%. Endocarditis developed in 1 patient. Freedom from aortic regurgitation is to date 100% at discharge, 6 months, and 1 year postimplant. Reoperation of the aortic valve/root was not required in any patient. Six-month mean transvalvular gradients for 21-29 mm valves were 12.8, 8.8, 5.3, 4.9, and 4.7 mm Hg, respectively. CONCLUSIONS: Aortic root replacement with the Toronto root is safe and provides superb transvalvular hemodynamics with freedom from anticoagulation. The Toronto root seems widely applicable for all types of aortic root pathology and these early data offer very encouraging results. Long-term follow-up is required. PMID- 15336993 TI - Repair of ischemic mitral regurgitation does not increase mortality or improve long-term survival in patients undergoing coronary artery revascularization: a propensity analysis. AB - BACKGROUND: The purpose of this study was to compare operative mortality and midterm outcome of patients with ischemic mitral regurgitation (MR) undergoing either coronary artery bypass grafting (CABG) alone or CABG with mitral valve (MV) repair. METHODS: From 1996 to 2001, 51 consecutive patients underwent CABG with MV repair for ischemic MR. All patients in this group were matched to similar patients with ischemic MR undergoing CABG alone during the same 6-year period using propensity analysis (considering 24 covariates, including severity of MR and New York Heart Association [NYHA] class). RESULTS: Propensity score matching yielded 51 closely matched control patients. Preoperative MR severity was 3+ or 4+ in 94% of CABG with MV repair and 96% of CABG-alone patients, and 86% of patients in each group were NYHA class III or IV. Operative mortality was 3.9% +/- 2.8% in both groups. Survival was also similar between CABG with MV repair and CABG alone at 1 year (84% +/- 5% versus 82% +/- 5%) and 3 years (70% +/- 7% versus 71% +/- 7% (p = 0.43). Among survivors, NYHA class improved at follow-up (50 +/- 20 months) from 3.4 +/- 0.7 to 1.7 +/- 1.0 for CABG with MV repair (p < 0.001) and from 3.4 +/- 0.7 to 1.8 +/- 1.0 for CABG alone (p < 0.001). CONCLUSIONS: Operative mortality, midterm survival, and late functional class were similar between two well-matched groups of patients undergoing CABG for ischemic MR, differing only in the addition of MV repair. Whereas MV repair can be added safely to CABG in this group of high-risk patients without increasing mortality, its impact on late survival and functional class may be limited. PMID- 15336994 TI - Consequence of atrial fibrillation and the risk of embolism after percutaneous mitral commissurotomy: the necessity of the maze procedure. AB - BACKGROUND: We sought to investigate the incidence and risk of embolism after percutaneous balloon mitral commissurotomy (PMC) and delineated the necessity of the maze procedure in patients with atrial fibrillation (AF). METHODS: Clinical records of 326 patients, who underwent PMC between April 1988 and March 2003, and 162 patients, who underwent mitral valve surgery (MVS) combined with the maze procedure (MVS/maze) in the same period, were reviewed. We examined the consequence of cardiac rhythm and the incidence and risk of embolism after PMC. Next our current criteria for indication of the maze procedure, which included duration of AF less than 10 years, voltage of f wave on electrocardiography greater than 0.1 mV, cardiothoracic ratio less than 70%, and left atrial dimension less than 70 mm, were applied to all patients. These criteria were satisfied by 190 patients of PMC (PMC group) and 114 patients of MVS/maze (MVS/maze group) and the early and late results of the two groups were compared. RESULTS: Thirty-eight patients suffered from embolic complications after PMC. Of these 33 patients experienced AF before intervention. The actuarial embolism-free rate at 10 years was 81.9% in patients with AF before PMC and 92.9% in patients with sinus rhythm before PMC (p = 0.01). Univariate predictors of embolism after PMC included previous embolic history (p = 0.01), AF (p = 0.01), pressure gradient (p = 0.01), age (p = 0.03), and mitral valve area (p = 0.04). Multivariate analysis identified AF as the independent predictor of embolism in the late follow-up period (p = 0.03). In a comparative study of the selected patients the actuarial AF-free rates after MVS/maze were 89.9% at 1 year and 85.7% at 5 years and were significantly higher than those of 17.3% at 1 year and 4.2% at 5 years after PMC (p < 0.0001). The actuarial embolism-free rate in the MVS/maze group was 98.0% at 5 years and was significantly higher than that of 84.8% at 5 years and 82.4% at 10 years in the PMC group (p = 0.01). CONCLUSIONS: PMC alone is not sufficient with regard to sinus rhythm recovery and prevention of embolism. Surgical treatment concomitant with the maze procedure may be beneficial for patients with AF. PMID- 15336996 TI - Results of mitral surgery in octogenarians with isolated nonrheumatic mitral regurgitation. AB - BACKGROUND: Increasing numbers of elderly patients are now referred for mitral valve operations. It has been unclear whether the results offset the risk of intervention in this patient population. METHODS: We obtained clinical follow-up through May 2002 of 59 patients 80 years or older who underwent first-time isolated mitral valve repair (46 patients) or replacement (13 patients) for nonischemic, nonrheumatic mitral regurgitation from January 1990 to June 2000. The mean duration of follow-up was 68 +/- 33 months. Observed survival was compared with the expected survival of persons of the same age and gender in the general population. RESULTS: Preoperatively 79% of patients were in New York Heart Association (NYHA) class III-IV. Operative mortality was 1.7%. Overall 1- and 5-year survival was 89% and 61%. One- and 5-year freedom from thromboembolic complications in hospital survivors was 97% and 84%. One- and 5-year freedom from heart-related hospitalization in hospital survivors was 89% and 78%. There were no reoperations. Twenty-nine patients underwent an echocardiographic follow-up; 31% of them exhibited moderate or more regurgitation. Of 37 surviving patients at follow-up, 78% were in NYHA functional class I-II. No statistically significant difference was noted between the observed survival postoperatively and the expected survival of persons of the same age and gender in the general population. In a univariate analysis, only preoperative left ventricular ejection fraction greater than 40% was significantly associated with freedom from late heart-related mortality (95% confidence interval 62%-92%, p = 0.01) and with freedom from heart-related hospitalization (95% CI 68%-95%, p < 0.01). CONCLUSIONS: Native mitral valve surgery for isolated nonischemic, nonrheumatic disease in octogenarians resulted in a survival rate comparable with that of the general population. It also exhibited substantial improvement regarding the functional status of the patient. Reparative techniques did not result in a survival advantage compared with replacement but did prove to be a reliable approach. Surgery performed in an early stage, preceding the development of left ventricular dysfunction, was associated with an improved freedom from late cardiac complications. PMID- 15336998 TI - Extensive use of polytetrafluoroethylene artificial grafts for prolapse of posterior mitral leaflet. AB - BACKGROUND: There are an increasing number of reports concerning mitral valve repair by means of reconstruction of the chordae tendinae with expanded polytetrafluoroethylene (e-PTFE) sutures. However little information is available about extended application or results of this technique for extended prolapse of posterior mitral leaflets. METHODS: Between March 1994 and December 2000, 22 patients with moderate-to-severe mitral regurgitation (MR) as the result of a prolapse of posterior leaflets (age range, 39-73 years) underwent mitral valve repair by means of reconstruction of artificial chordae with 4-CV e-PTFE sutures without leaflet resection. Either Kay's suture or ring annuloplasty was also performed to correct annular dilatation in all patients. RESULTS: No operative death or late mortality was observed. Before discharge immediate postoperative echocardiography indicated less than moderate MR in 20 out of 22 patients. The follow-up was complete in all cases by clinical examination and serial echocardiograms and the median follow-up period was 87 months (range 24-108). There were two failures that required reoperation because of unsuccessful repair and worsening MR (elongation of the anchored side of the papillary muscle). When the reoperated patients were excluded from the follow-up data, the degree of MR, estimated by echocardiography that was performed at a recent follow-up period, was nonexistent in 6 patients, trivial in 10 patients, and mild in 4 patients. The systolic and diastolic dimensions of the left ventricle decreased significantly (p < 0.01). CONCLUSIONS: Replacement of the artificial chordae was not complicated and seemed to preserve favorable relationships among leaflet tissues, chordae, and papillary muscles. We therefore suggest that the extensive use of PTFE artificial chordae seems to be a promising procedure regarding the repair of many kinds of mitral lesions causing MR. PMID- 15336999 TI - Selecting patients with mitral regurgitation and left ventricular dysfunction for isolated mitral valve surgery. AB - BACKGROUND: American College of Cardiology/American Heart Association (ACC/AHA) Guidelines state that patients with an ejection fraction (EF) of 30% or less should not undergo mitral valve replacement for mitral regurgitation (MR). We sought to establish, using a national cardiac surgery database, whether patients with left ventricular dysfunction may safely undergo mitral valve surgery for MR, and if so, which ones. METHODS: We queried the Society of Thoracic Surgeons (STS) National Database to identify patients who had isolated mitral valve replacement or repair for MR between 1998 and 2001. Mortality and morbidity outcomes were compared by EF category (< or = 30% vs > 30%), and observed mortality compared by EF group, stratified by predicted risk for mortality. A classification and regression tree (CART) model was then used to determine which patient characteristics contributed most to designate the high-risk patient. RESULTS: Of the 14,582 patients who had mitral valve surgery, 727 had an EF of 30% or less and 13,855 had an EF of more than 30%. Observed mortality rates were higher for patients with an EF of 30% or less (5.4% vs 3.1%). However, for low-risk to medium-risk patients, mortality rates remained fairly constant across levels of EF. Mortality is notably increased in the high-risk patients (predicted risk > 10%). A classification tree identifies three key characteristics for high risk: age more than 75 years, renal failure, and emergent or salvage procedure. CONCLUSIONS: When the predicted mortality risk is less than 10%, EF has minimal impact on operative mortality for mitral regurgitation. In contrast to the ACC/AHA Guidelines, our data show that operative risk for mitral valve surgery is not prohibitive for most patients with ventricular dysfunction. PMID- 15337000 TI - Mitral and tricuspid valve repair in patients with previous mediastinal radiation therapy. AB - BACKGROUND: The purpose of this study was to evaluate outcomes of mitral and tricuspid valve repair after mediastinal radiation therapy. METHODS: From 1976 to 2001, 22 patients (mean age 61 +/- 14 years) underwent mitral (n = 14), tricuspid (n = 6), or both (n = 2) valve repairs 15 +/- 9 years after mediastinal radiation therapy. Concomitant procedures included coronary artery bypass graft, 11 patients; valve replacement, 6 patients (4 aortic, 3 mitral, 1 tricuspid, and 1 pulmonary); and pericardiectomy, 4 patients. RESULTS: Total follow-up was 82.5 patient-years (mean 3.7 +/- 3.3 years). Early mortality was 3 patients. There were 7 late deaths, 4 of which were of cardiovascular origin. Of the 19 early survivors, 2 required subsequent valve replacements, and 1 required cardiac transplantation 3.4 +/- 2.8 years after valve repair. One patient died after reoperation. In 4 patients who did not undergo reoperation, echocardiographic examinations showed progressive deterioration of their repaired valve function. Overall survival, freedom from cardiac death, and freedom from valve reoperation or cardiac transplantation at 5 years for early survivors was 66%, 85%, and 88%, respectively. New York Heart Association functional class at follow-up was I or II in 8 of the 12 late survivors. CONCLUSIONS: Functional status was good in two thirds of late survivors. However, severe dysfunction of the repaired valve developed in 32% of early survivors and 16% required further surgery. Valve repair is technically feasible in selected patients after mediastinal radiation therapy; however, the limited durability of repairs after mediastinal radiation in this series suggests that valve replacement might be preferable. PMID- 15337001 TI - Clinical outcomes with the Hancock II bioprosthetic valve. AB - BACKGROUND: The Hancock II bioprosthetic valve, which was first introduced to clinical use in 1978, differs from its predecessor in several ways. This study was designed to evaluate the durability and outcomes with this valve in patients who had isolated aortic or mitral valve replacements. METHODS: From 1991 to 1999, 459 patients underwent aortic valve replacement and 138 patients underwent mitral valve replacement with the Hancock II bioprosthesis (Medtronic Inc, Minneapolis, MN). The mean age was 73.2 +/- 0.4 and 72.6 +/- 0.8 years in the aortic and mitral groups, respectively. Most patients were in New York Heart Association Class III or IV (50% aortic group and 69% mitral group) and concomitant coronary artery bypass was performed in 49.4% and 52.8% of patients, respectively. Patients were assessed annually and follow-up was up to 129 months in the aortic group and 100 months in the mitral group. RESULTS: At 8 years, actuarial survival was 52% +/- 5% in the aortic group and 57% +/- 8% in the mitral group. Furthermore, the actuarial freedom from structural failure necessitating reoperation was 99% +/- 0.5% in the aortic group and 98% +/- 2% in the mitral group, and the actuarial freedom from repeat valve surgery due to all causes was 97% +/- 2% and 96% +/- 2%, respectively. Actuarial freedom from thromboembolic events was 89% +/- 2% in the aortic group and 90% +/- 5% in the mitral group. CONCLUSIONS: The Hancock II valve has excellent midterm durability and clinical performance in older patients. PMID- 15337002 TI - Determinants of early and late outcome for reoperations of the proximal aorta. AB - BACKGROUND: The purpose of this study was to investigate the cause of ascending aorta and aortic arch reoperations and to identify determinants of early and late outcome. METHODS: Between January 1991 and March 2003 we repaired aneurysms of the proximal aorta in 597 patients. Of these patients, 104 had reoperations for replacement of the ascending aorta, aortic root, or transverse aortic arch. Previous surgery was defined as any previous cardiac or proximal aortic repair. Median age was 60 years, and 29 of the patients (28%) were female. Indications for reoperation and replacement of the proximal aorta included acute type A dissection in 6 patients (5.8%), aneurysm with chronic dissection in 60 (57.7%), progression of aneurysm in 23 (22.1%), infection in 12 (1.5%), inflammatory disease in 2 (1.9%), and atheromatous disease in 1 (1.0%). Reoperations included aortic root replacement in 20 patients (19.2%), total arch replacement with elephant trunk in 28 (26.7%), ascending and proximal arch in 39 (37.5%), and ascending aorta in 27 (26.0%). The median interval between operations was 69 months. Retrograde cerebral perfusion was used in 80 (77%) cases. RESULTS: Chronic dissection was the most common indicator for reoperation in our population, followed by progression of aneurysm and infection. Thirty-day and in hospital mortality was 13.5% (14 of 104) and 15.4% (16 of 104), respectively. Chronic obstructive pulmonary disease, renal dysfunction, and increased pump time were risk factors for mortality. Median follow-up was 5.02 years. Eight patients died during that period. Estimated survival at 1, 5, and 10 years was 83%, 80%, and 62%, respectively. Freedom from second proximal reoperations was 97.1% (10 of 104). Freedom from subsequent distal thoracic aortic repair was 84.6% (8 of 104). CONCLUSIONS: Reoperations of the ascending aorta and aortic arch can be performed safely with good long-term results. Patients with previous proximal aortic dissection repair need long-term surveillance. Renal dysfunction and chronic obstructive pulmonary disease must be carefully considered before reoperations of the proximal aorta. PMID- 15337003 TI - Magnetic resonance angiographic localization of the artery of Adamkiewicz for spinal cord blood supply. AB - BACKGROUND: The purpose of this study was to determine whether the artery of Adamkiewicz (ARM) can be detected by magnetic resonance angiography and to determine the usefulness of preoperative magnetic resonance angiography evaluation of the ARM. METHODS: Between April 2000 and December 2003, 120 patients underwent magnetic resonance angiography for detection of the ARM. The morphology of the anterior spinal artery at the ARM junction, as revealed by magnetic resonance angiography, in 99 patients in whom ARM was preoperatively detected was classified into the following three types: noncontinuation of the anterior spinal artery above the ARM junction (type A), continuation of the anterior spinal artery above and below the ARM junction (type B), and noncontinuation of the anterior spinal artery below the ARM junction (type C). RESULTS: The ARMs were detected in 99 (83%) of 120 patients, and from a total of 110 ARMs 105 (95%) originated from intercostal arteries branching from the left side and 94 (86%) originated between Th9 and Th11. Two ARMs were found in 11 (11%) of 99 patients in whom ARMs were detected. In 107 patients, who underwent magnetic resonance angiography to reveal the morphology of the anterior spinal artery at the ARM junction, the patterns of the anterior spinal artery were type A in 59 patients (55%), type B in 21 patients (20%), type C in 3 patients (3%) and not classified in 24 patients (22%). No spinal cord injury occurred in patients in whom the ARM had been preoperatively detected. CONCLUSIONS: Preoperative detection of the ARM is possible by magnetic resonance angiography and is very useful for reducing the incidence of ischemic injury of the spinal cord. PMID- 15337005 TI - Type A acute aortic dissection: immediate and mid-term results of emergency aortic replacement with the aid of gelatin resorcin formalin glue. AB - BACKGROUND: The aim of this study was to assess the mid-term results of operation for type A acute aortic dissection with the aid of gelatin resorcin formalin glue. METHODS: Emergency operation was carried out in 84 patients during the last 8 years. Fifty-five patients (65.5%) had mild-to-moderate aortic regurgitation. Gelatin resorcin formalin glue was applied to both the proximal and distal aortic stumps. We evaluated the presence of aortic regurgitation and the patency of the distal false lumen at the time of this study. The survival and reoperation-free rates were also assessed. In case of late reoperation, aortic wall samples of the glued area were examined histologically. RESULTS: Ascending to hemiarch replacement were performed in 71 patients (84.5%). Total aortic arch and root replacement were required in 13 and 7 patients, respectively. Overall hospital mortality was 6.0% (5 patients). Late death was observed in 12 patients (14%). Reoperation for redissection in the aortic root, development of aortic regurgitation, and enlargement of the distal false lumen occurred in 1, 3, and 1 patient, respectively. Histologic examination showed no evidence of infiltration of inflammatory cells in the glued area. Computed tomography scan revealed a patent distal false lumen in 8 (14%) of 58 patients. Echocardiography detected moderate aortic regurgitation in 2 patients. The actuarial survival rate at 1, 5, and 8 years was 85.5%, 80%, and 60.0%, respectively. The reoperation-free rate at 8 years was 89%. CONCLUSIONS: The results of emergency aortic replacement with gelatin resorcin formalin glue have shown reasonable early and late mortality and reoperation rates. There was no histologic evidence of adverse tissue reactivity by gelatin resorcin formalin glue. PMID- 15337007 TI - Reduction aortoplasty and external wrapping for moderately sized tubular ascending aortic aneurysm with concomitant operations. AB - BACKGROUND: Replacement of the ascending aorta with a prosthetic graft is the preferred surgical procedure for an ascending aortic aneurysm. The choice of external wrapping of the aorta is a simple, fast, and effective method for moderately sized ascending aortic aneurysms with concomitant operations. In this study, we present the midterm results of 62 patients undergoing reduction aortoplasty with external wrapping and different cardiac procedures. METHODS: The study group consisted of 48 male and 14 female patients with a mean age of 59.3 +/- 6.0 years. Measurement of the ascending aorta diameters was obtained at three points: before surgery, during the early postoperative period, and during the follow-up. The mean preoperative aortic diameter was 52.7 +/- 0.5 mm. In all patients, the ascending aortic aneurysm was repaired by reduction aortoplasty with external wrapping. RESULTS: Mean follow-up time was 39.6 +/- 18.0 months. There was only one mortality (1.6%) as a result of septic multiple-organ failure and no major surgical complications in the 30-day postoperative period. Reduction aortoplasty of the ascending aorta with external wrapping resulted in a significant reduction of the ascending aorta in all patients (p = 0.000). There was an increase in the mean aortic diameter during the follow-up period (p = 0.000). Although this increase was statistically significant, all measurements of the follow-up period were still within normal range. CONCLUSIONS: External wrapping of the aorta offers excellent results with very low mortality and morbidity, and it can be regarded as a safe and effective method for the treatment of ascending aortic aneurysm in selected patients. However, the patients should be carefully monitored for redilatation after the procedure. PMID- 15337008 TI - Transesophageal echocardiography for detecting extrapulmonary thrombi during pulmonary embolectomy. AB - BACKGROUND: Pulmonary embolectomy is an evolving surgical procedure for the treatment of severe pulmonary embolism. In addition to removing pulmonary thromboemboli, the achievement of optimal results also requires identification and extraction of intrathoracic, extrapulmonary thromboemboli from the right atrium, right ventricle, and the superior or inferior vena cava. Otherwise, these thromboemboli may become the source of recurrent pulmonary embolism. Intraoperative transesophageal echocardiography is frequently used during pulmonary embolectomy as a guide for the surgeon and a monitor of cardiac performance. However, its utility for detecting concurrent intrathoracic, extrapulmonary thromboemboli has not been thoroughly investigated. METHODS: The study population included 50 consecutive patients undergoing emergent pulmonary embolectomy. Results from preoperative diagnostic studies including transthoracic echocardiography; ventilation/perfusion scan, pulmonary angiography, and computed tomography were reviewed, along with the intraoperative echocardiographic reports for the presence of intrathoracic extrapulmonary thromboemboli. In addition, the surgical operative notes were consulted to determine the impact of intraoperative transesophageal echocardiography on surgical decision making. RESULTS: Extrapulmonary intrathoracic thromboemboli were identified preoperatively in 8 patients (16%). Intraoperatively, thromboemboli were identified in 5 additional patients by transesophageal echocardiography, adding to a total of 13 patients with extrapulmonary thrombi (26%). Intraoperative transesophageal echocardiography provided the sole source of evidence that directed surgical management in 10% (5 of 50) of all patients undergoing pulmonary emboletcomy. CONCLUSIONS: Intraoperative transesophageal echocardiography identified intrathoracic, extrapulmonary thomboemboli in 26% of patients undergoing pulmonary embolectomy, resulting in an alteration of surgical management in 10% of patients. These findings support the critical role of intraoperative echocardiography during pulmonary embolectomy. PMID- 15337010 TI - Left anterior descending coronary endarterectomy: early and late results in 196 consecutive patients. AB - BACKGROUND: With advances in percutaneous coronary interventions, many patients now referred for coronary artery bypass grafting have diffuse coronary artery disease. We undertook this retrospective study to determine whether left anterior descending (LAD) coronary endarterectomy is a safe and effective long-term adjunct to coronary artery bypass grafting in patients who cannot otherwise be completely revascularized. METHODS: Between January 1992 and March 2000, 196 of 7,633 (2.5%) consecutive patients underwent LAD coronary endarterectomy with coronary artery bypass grafting. Median age was 67 years (range, 33 to 97 years), 101 patients (52%) had unstable angina, and 182 (93%) were in New York Heart Association class III or IV. Thirty-three patients (17%) had ongoing myocardial infarction; another 17 (9%) had myocardial infarction less than 1 month. Thirty patients (15%) required intraaortic balloon pump preoperatively and 19 (10%) were reoperations. RESULTS: All patients underwent LAD endarterectomy with coronary artery bypass grafting to the LAD. The left internal mammary artery was grafted to the LAD in 151 patients (77%), and 46 of 151 (30%) of these required an additional vein patch to the endarterectomized bed. Concomitant valve procedures were performed in 8 (4%) patients. Overall hospital mortality was 3% (6 of 196). Perioperative myocardial infarction in the LAD territory was 3%. One-year survival was 94% (95% confidence interval, 90% to 97%), whereas 5-year survival was 74% (95% confidence interval, 66% to 80%). Freedom from cardiac events (angina, myocardial infarction, congestive heart failure, percutaneous coronary interventions) was 90% (95% confidence interval, 84% to 94%) at 1 year and 84% (95% confidence interval, 75% to 90%) at 5 years. CONCLUSIONS: Despite the presence of diffuse coronary artery disease, coronary artery bypass grafting with LAD endarterectomy offers excellent results with very low hospital mortality and morbidity, and favorable long-term survival. PMID- 15337012 TI - Transmyocardial revascularization using an XeCl excimer laser: results of a randomized trial. AB - BACKGROUND: CO2 and holmium:yttrium aluminum garnet (YAG) transmyocardial laser revascularization (TMLR) are used to treat patients with refractory angina. A randomized trial to investigate the efficacy and safety of XeCl excimer TMLR was performed. METHODS: Thirty patients with refractory angina were randomized in pairs to excimer TMLR or maximal medication. We assessed angina, quality of life (QOL), exercise time, myocardial perfusion, and ventricular wall motion at base line and at 3, 6, and 12 months after TMLR. RESULTS: TMLR patients manifested a significantly better outcome with respect to angina class and quality of life. One TMLR patient died perioperatively versus none in the control group. After TMLR angina decreased from class 3.8 +/- 0.4 at base line to 1.9 +/- 0.9 at 12 months versus 3.9 +/- 0.3 to 3.7 +/- 0.6 in the control group, respectively (p = 0.000001). At 12 months a decrease of greater than or equal to two angina classes was indicated in 11 out of 14 TMLR patients versus none in the control group (p = 0.00001). Improved myocardial perfusion or exercise time was not indicated despite a small decrease in reversible wall motion abnormality score. CONCLUSIONS: Excimer TMLR significantly relieves angina and improves QOL without evidence of improved cardiac perfusion or function. PMID- 15337014 TI - Detection of anti-hsp70 immunoglobulin G antibodies indicates better outcome in coronary artery bypass grafting patients suffering from severe preoperative angina. AB - BACKGROUND: Recent findings indicate that molecular chaperones actively participate in myocardial cytoprotection. Moreover, ischemic tolerance can be induced in humans by brief ischemic events. Therefore, we investigated patients with severe angina attacks before coronary artery bypass grafting. We focused on appearance of anti-hsp70 antibodies as an immunologic response to heat shock protein induction by ischemia followed up by hemodynamic measurements perioperatively. We correlated these clinical findings with the presence of antibodies against hsp70 and the antioxidative capacity of patients' sera. METHODS: Thirty-five consecutive patients with coronary artery disease scheduled for coronary artery bypass grafting were included. Seventeen patients had severe angina, and 18 patients suffered from chronic stable angina preoperatively. In the patients' sera, antibodies against hsp70 were detected by enzyme-linked immunosorbent assay, and antioxidative capacity was detected using the chromogen assay. Cardiac output and pulmonary capillary wedge pressure were measured using a thermodilution catheter. We also evaluated C-reactive protein and creatine kinase MB isoenzyme, and performed a conventional leukocyte count. RESULTS: The sera of the 17 patients with severe angina attacks before surgery contained antibodies against hsp70 and a low antioxidative capacity. The interval between a severe angina attack and anti-hsp70 antibody titer are inversely correlated. These patients had better cardiac output and lower pulmonary capillary wedge pressure values after surgery. CONCLUSIONS: Severe angina before cardiac surgery coincided with an improved outcome as measured by hemodynamic variables as compared with chronic stable angina. This finding correlated significantly with a low antioxidative capacity and the presence of antibodies against hsp70. These pathophysiologic mechanisms might therefore play a role in myocardial protection. PMID- 15337016 TI - Effect of older donor age on risk for mortality after heart transplantation. AB - BACKGROUND: Despite the increasingly common use of donor hearts at least 50 years of age, controversy still remains regarding long-term outcome. Our goal was to determine if older donor age is associated with an increased risk of mortality and specifically if the use of donor hearts at least 50 years of age reduces survival. METHODS: We retrospectively studied records of all primary heart transplants performed between January 1990 and July 2002. Fifty-six patients who had received donor hearts at least 50 years of age were compared with 611 recipients of donor hearts less than 50 years of age. Clinicopathologic parameters were analyzed for their effect on mortality using the Cox proportional hazard model with calculation of hazard ratios (HR). Cut-point analysis of donor age was used to determine which donor age is associated with the greatest risk of mortality after transplant. RESULTS: Recipients of donor hearts at least 50 years of age were older (58.5 years +/- 7.0 vs 53.2 +/- 11.6; mean +/- standard deviation [SD]; p < 0.0001), suffered more often from ischemic cardiomyopathy (69% vs 50%, p = 0.01), and experienced a longer waiting time (192.2 days +/- 301.0 vs 138.6 +/- 190.8, p < 0.0001). Donor hearts at least 50 years of age (age 54.1 +/- 3.5 years) were more often female (50% vs 34%, p = 0.03), died less often of "head trauma" (9% vs 42%, p < 0.0001), and exhibited fewer cytomegalovirus (CMV) mismatches (29% vs 39%, p = 0.04) than donor hearts less than 50 years of age (age 26.8 +/- 12.3 years). Multivariate predictors of mortality were rejection index (HR 1.90 per unit [rejections/100 survival days], p < 0.0001), donor age (HR 1.16 per 10-year increment, p = 0.002), and recipient age (HR 1.24 per 10-year increment, p = 0.04). Recipients of donor hearts at least 50 years of age had reduced 1-year and 5-year survival ([65.7% vs 81.7%, p < 0.05] and [48.3% vs 68.4%, p < 0.05], respectively), as well as a higher proportion of deaths occurring within 1 month of transplant (41% of total deaths vs 23%, p = 0.06). Cut-point analysis indicated the characteristic of donor age of at least 40 years (categorical variable) to predict mortality with the same degree of fit as age used as a continuous variable. CONCLUSIONS: Although we observed a substantial reduction in survival among patients who were allocated donor hearts at least 50 years of age, this difference was not solely attributable to the categorical variable of donor age 50 in this group. Donor age as a continuous variable, however, was determined to be a notable predictor of survival and use of the donor age cut-point of 40 years (categorical variable) allowed risk stratification with similar accuracy. The use of a donor age cut point of 40 years may be a useful clinical criterion for graft-related risk assessment. PMID- 15337017 TI - Changes in right ventricular dimensions and performance after passive cardiac containment. AB - BACKGROUND: Previous studies have shown that the cardiac support device (CSD) improves left ventricular structure and function in patients with heart failure by preventing further cardiac enlargement. The aim of this study was to identify effects on the right ventricle (RV). METHODS: Ten male patients with idiopathic dilated cardiomyopathy underwent electron-beam computed tomographic (CT) examination within 1 month before, and 6 to 9 months after CSD implantation. The RV end-diastolic and end-systolic volumes (EDV, ESV) and diameters (EDD, ESD), stroke volume (SV), ejection fraction (EF), total and forward RV output (RVO, fRVO), and tricuspid regurgitation fraction (TRF) were calculated. RESULTS: The EDV measurements decreased from 182.1 +/- 49.6 to 137.5 +/- 37.0 mL, ESV from 114.8 +/- 47.0 to 68.3 +/- 23.8 mL, EDD from 48.2 +/- 6.6 to 41.6 +/- 7.1 mm, and ESD from 39.6 +/- 6.9 to 32.7 +/- 6.5 mm (p < 0.05 for each). Ejection fraction increased from 38.5 +/- 8.9 to 52.0% +/- 7.7% and fRVO from 4.0 +/- 0.8 to 4.6 +/ 1.1 L/min (each with p < 0.05). TRF decreased from 18.2 +/- 14.1 to 10.4% +/- 13.5%, whereas SV and RVO remained nearly unchanged. Postoperatively, RV volumes, EF, and fRVO were not different from 15 age- and gender-matched normal control patients. CONCLUSIONS: Implantation of a CSD leads to a decrease in RV size and improved RV performance. These data together with the results of previous studies demonstrating improved left ventricular structure and function confirm the biventricular nature of recovery with the CSD. PMID- 15337018 TI - Sleep deprivation does not affect operative results in cardiac surgery. AB - BACKGROUND: There has been an increasing trend towards the mandatory reduction in work hours for physicians because of the fear that sleep-deprived (SD) surgeons are more prone to make mistakes. We hypothesized that sleep deprivation would not be associated with increased morbidity or mortality in cardiac operations. METHODS: A retrospective review was done of all cases performed by all attending cardiac surgeons from January 1994 to April 2003. Complication rates of cases performed by SD surgeons were compared with cases done when the surgeons were not sleep-deprived (NSD). A surgeon was deemed sleep deprived if he or she performed a case the previous evening that started between 10:00 pm and 5:00 am, or ended between the hours of 11:00 pm and 7:30 am. RESULTS: A total of 6,751 cases were recorded in the Society of Thoracic Surgeons database over the 9-year period examined. Of these, 339 cases (5%) were performed by SD surgeons, and 6,412 (95%) cases were performed by NSD surgeons. Mortality rates for coronary artery bypass operations showed no significant differences (1.7% [SD = 4/223] vs 3.1% [NSD = 133/4206)] p = 0.34). Operative (p = 0.47), pulmonary (p = 0.60), renal (p = 0.93), neurologic (p = 0.11), and infectious (p = 0.87) complications of all cases also failed to show any statistically significant differences in any group. Perfusion times, cross-clamp times, and the use of blood products were also similar between groups. CONCLUSIONS: Sleep deprivation does not affect operative morbidity or mortality in cardiac surgical operations. These data do not support a need for work hour restrictions on surgeons. PMID- 15337019 TI - Postoperative sternal dehiscence in obese patients: incidence and prevention. AB - BACKGROUND: Obesity has been identified as the single most important risk factor for postoperative sternal infection in coronary bypass surgery patients. It is also a major risk factor for sternal dehiscence, with or without infection, for any type of cardiac operation. We assessed whether prophylactic measures could prevent this complication. METHODS: Two studies were conducted. In study A, 3,158 heart surgery patients were analyzed at 3 cardiac units. Obesity was defined as body mass index (BMI) more than 30. Group I (1,253 obese [39.7%]) was compared with group II (1,905 nonobese [60.3%]). Sternal closure was done at the surgeon's preference: (a) plain wires through and through the bone; (b) peristernal figure of-eight wires; or (c) peristernal method, using stainless-steel cables. In study B, 123 obese patients were prospectively divided into 2 subgroups. Group B-1 (54 patients) underwent lateral prophylactic sternal reinforcement before placement of peristernal wires. Group B-2 (69 patients) had standard sternal closure, as in study A. RESULTS: In study A, group I had 81 dehiscences (6.46%); 78 also suffered deep sternal infection and mediastinitis (96%). Despite treatment, dehiscence recurred in 13, and mortality was 38.4%. In group II nonobese patients, 31 dehisced (1.6%, p = 0.000), with no mortality. In study B, group B-1 (54) had 0% dehiscence versus group B-2 (69) with 6 dehiscences (8.7%). CONCLUSIONS: In our study, the rate of obesity is high ( approximately 40%). Sternal dehiscence is real when the BMI is more than 30 (6.46%), and has high morbidity and mortality. Prophylactic sternal reinforcement seems to prevent this complication. PMID- 15337020 TI - Ventricularization of the atrialized chamber: a concept of Ebstein's anomaly repair. AB - BACKGROUND: We report results of a technique of Ebstein's anomaly repair by creating a predominantly monocuspid valve with simultaneous ventricularization of the atrialized right ventricular (aRV) chamber. METHODS: Between March 1993 and April 2003, Ebstein's anomaly repair by valvuloplasty with combined ventricularization was performed in 23 patients aged 13.6 (4.1-52.6) years presenting with tricuspid valve regurgitation (TVR) (I degrees, n = 1; II degrees, n = 3; III degrees, n = 13; IV degrees, n = 6). Valvuloplasty consisted of creating a predominantly monocuspid valve at the level of the anatomical atrioventricular junction resulting in a ventricularization of the atrialized chamber. Postoperatively all survivors were examined regularly with an actual prospective evaluation. RESULTS: One early death (4.4%) occurred and was caused by right heart failure. Follow-up was 4.6 (0.5-10.9) years. Important recurrent atrioventricular valve regurgitation caused by rupture of fixation sutures occurred in 3 patients (13%), necessitating reintervention at 3 (0.03-4) months (revalvuloplasty, n = 2; TV replacement, n = 1). One patient presenting with hypoplastic right ventricle with consecutive right heart failure underwent creation of a total cavopulmonary connection at 10 months. At present all patients are doing well. Actual echocardiographic examination revealed significant improvement of right atrioventricular valve regurgitation (p < 0.0001) and favorable restoration of RV geometry and function. CONCLUSIONS: This technique of Ebstein's anomaly repair with ventricularization of the atrialized chamber provides excellent results regarding right atrioventricular valve function and leads to a favorable restoration of RV geometry and function. PMID- 15337022 TI - Anatomic reconstruction for recurrent aortic obstruction in infants and children. AB - BACKGROUND: Patients undergoing operative repair of aortic obstruction are at a lifelong risk of recurrent obstruction, and there is controversy regarding the optimal surgical technique. We have used an alternative strategy for recurrent aortic obstruction, typically involving anatomic reconstruction by means of a median sternotomy, and describe our techniques and results. METHODS: Twenty-one patients presented with recurrent aortic arch obstruction. Mean age and weight were 7.8 +/- 5.4 years (range, 0.21 to 15.2 years) and 30.6 +/- 21.8 kg (range, 3.6 to 90 kg), respectively. Recurrence involved the aortic arch to some degree in each case, as the mean preoperative transverse aortic arch z score was -2.9 +/ 1.6 (range, -7.0 to 0.1). Thoracotomy was possible in 2 patients, using re resection with end-to-end anastomosis (n = 1) and patch aortoplasty (n = 1). The remaining 19 patients required median sternotomy, cardiopulmonary bypass, and deep hypothermic circulatory arrest for complete relief of obstruction by aortic arch advancement (n = 10), patch aortoplasty (n = 8), or interposition grafting (n = 1). RESULTS: There was 1 hospital death. Invasive blood pressure monitoring revealed no residual arm-to-leg gradient in 19 patients and a 20-mm Hg gradient in 2 patients. There have been no late deaths. No patients have undergone subsequent aortic intervention, and all are asymptomatic up to 85 months postoperatively. Two patients are currently followed with a 10-mm Hg arm-to-leg blood pressure gradient. CONCLUSIONS: Anatomic reconstruction for recurrent aortic obstruction can be safely accomplished in the majority of patients. We favor median sternotomy because of the ability of establishing cardiopulmonary bypass, the facility of anatomic reconstruction techniques, and the ability to repair concomitant cardiovascular lesions. PMID- 15337023 TI - Hemodynamic status after the Norwood procedure: a comparison of right ventricle to-pulmonary artery connection versus modified Blalock-Taussig shunt. AB - BACKGROUND: The aim of this study is to compare hemodynamic status, in particular systemic oxygen delivery, in patients undergoing a Norwood procedure with a right ventricle-to-pulmonary artery (RV-PA) versus a modified Blalock-Taussig (mBT) shunt. METHODS: From June 2000 to November 2003, 44 consecutive neonates with hypoplastic left heart syndrome underwent a Norwood procedure. The first 25 patients received an mBT shunt; the subsequent 19 an RV-PA shunt. Hemodynamic data, including mixed venous oxygen saturation, was determined during the first 48 hours after surgery. RESULTS: The mBT and RV-PA shunt patients had no significant differences in systemic oxygen saturation, mixed venous oxygen saturation, arteriovenous oxygen saturation difference, or oxygen excess factor during the first 48 hours. Mixed venous saturation declined to a nadir in both groups at 6 to 12 hours. The RV-PA patients had significantly higher diastolic and mean blood pressures, and lower systolic blood pressure. Mean heart rate, common atrial pressure, and inotrope score did not differ between the two groups. The RV-PA patients received higher fraction of inspired oxygen and minute ventilation to achieve partial pressures of arterial oxygen and carbon dioxide, and pH, similar to mBT patients. Durations of mechanical ventilation, intensive care unit stay, and hospital stay did not differ between mBT and RV-PA patients. Operative survival in the mBT versus RV-PA group was 20 of 25 (80%) versus 17 of 19 (89%; p = 0.7). CONCLUSIONS: Indicators of postoperative systemic oxygen delivery are equivalent in neonates who have undergone a Norwood procedure with an mBT or RV-PA shunt. Both mBT and RV-PA patients undergo similar declines in hemodynamic status 6 to 12 hours after surgery. Any advantages of one approach over the other lie in areas other than systemic oxygen delivery, such as resistance to physiologic insults, or preservation of ventricular function. PMID- 15337024 TI - Increased vascular endothelial growth factor in patients with cyanotic congenital heart diseases may not be normalized after a Fontan type operation. AB - BACKGROUND: To determine the change of serum concentration of vascular endothelial growth factor (VEGF) in patients with cyanotic congenital heart disease (C-CHD). METHODS: Patients comprised four groups: group A, 19 patients without cyanosis; group B, 24 patients with C-CHD; group C, 17 patients who had C CHD and underwent biventricular repair; and group D, 15 patients who had single ventricle and underwent a Fontan type operation. Blood samples were obtained from upper arm veins and serum VEGF was determined. We determined correlation between serum VEGF and arterial oxygen saturation and compared levels of serum VEGF among groups. In addition, age and hemodynamic variables derived from cardiac catheterization were analyzed in terms of correlation with serum VEGF. RESULTS: Serum VEGF significantly negatively correlated with arterial oxygen saturation (r = -0.62, p < 0.0001). Serum VEGF in B and D were significantly higher than those in A and C (median = 443 pg/mL in B and 302 pg/mL in D vs 180 pg/mL in A and 216 pg/mL in C, respectively). Serum VEGF was significantly negatively correlated with patient's age (p = 0.54, p < 0.04) in D, but significantly positively correlated with systemic venous pressure (p = 0.45, p < 0.01) in pooled data. CONCLUSIONS: Patients with C-CHD have increased serum VEGF in parallel with the degree of cyanosis. With biventricular repair, cyanosis and serum VEGF are normalized. However, with a Fontan type operation, cyanosis disappears but serum VEGF may not be normalized because of elevated venous pressure in association with younger age. PMID- 15337026 TI - Apoptosis-related mitochondrial dysfunction in the early postoperative neonatal lamb heart. AB - BACKGROUND: In the early postoperative period, the neonatal myocardium undergoes sparse apoptotic cell loss ( approximately 1% of myocytes). Because apoptosis is preceded by events associated with mitochondrial dysfunction, the fraction of myocytes with preapoptotic mitochondrial changes has important clinical implications (eg, postoperative myocardial dysfunction). My colleagues and I therefore hypothesized that postoperative apoptotic myocytes represent a tip of the iceberg, with more myocytes upstream with apoptosis-related mitochondrial dysfunction (ARMD). METHODS: Neonatal lambs underwent cardiopulmonary bypass, 60 minutes of cardioplegic arrest, and 6 hours of recovery (cardiopulmonary bypass with cardioplegic arrest [CPB+CP]; n = 5) and were compared with nonbypass controls (non-CPB; n = 5). Myocardium (left ventricle [LV] and right ventricle [RV]) was examined by using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining, electron microscopy, immunohistochemistry, Western blot, and isolated mitochondrial oxygen consumption measurement. RESULTS: TUNEL-positive nuclei and electron microscopy confirmed mitochondrial structural changes were more common in CPB+CP than non CPB myocardium and were more common in the LV than RV (p = 0.0016). Bax (a proapoptotic mediator) translocated from the cytosol to the mitochondria (LV > RV; p < 0.05). Immunohistochemistry demonstrated diffuse mitochondrial loss of cytochrome c that was consistent with outer mitochondrial membrane permeabilization (LV > RV > non-CPB). Permeabilization was further demonstrated by augmentation of oxygen consumption in isolated mitochondria after administration of exogenous cytochrome c. The mitochondrial oxygen consumption boost was 57% for CPB+CP:LV; 23% for CPB+CP:RV; and 18% and 17% for non-CPB:LV and non-CPB:RV, respectively (p < 0.01, CPB+CP:LV vs other groups). CONCLUSIONS: ARMD is much greater than the prevalence of TUNEL-positive myocytes in postoperative neonatal myocardium. Greater LV vulnerability may represent a relationship between increased afterload and ARMD. These changes are consistent with the early postoperative myocardial dysfunction commonly reported after neonatal cardiac operations. PMID- 15337027 TI - Myocardial and pulmonary effects of aqueous oxygen with acute hypoxia. AB - BACKGROUND: The purpose of this paper was to evaluate myocardial and pulmonary effects of aqueous oxygen (AO) delivered directly into the pulmonary circulation in acute hypoxia. METHODS: Six calves (2 months old, 68.0 +/- 2.2 kg) after general anesthesia, mechanical ventilation, and median sternotomy underwent total right heart bypass using fixed flow with continuous pressure and blood gas measurements in carotid and femoral arteries, left atrium, the coronary sinus and PA. Measurements of systemic and PA pressures and O2 saturations; myocardial O2 atrioventricular (AV) differences; and O2 extraction were made. After base line measurements, hypoxic ventilation reducing the mean arterial PO2 from 277 +/- 102 mm Hg to 47 +/- 4 mm Hg (p < 0.0005) was maintained for 30 minutes. Without changes in the hypoxic ventilation (mean arterial PO2 = 49 +/- 11 mm Hg) 3 ml/min of hyperbaric aqueous oxygen (AO = oxygen diluted in saline solution) was administered into the PA for 30 minutes. Pulmonary blood flow was maintained during the entire experiment (3.7 +/- 0.3 L/min). RESULTS: Hypoxic ventilation significantly raised (p < 0.05) the systolic (30 +/- 7 vs 21 +/- 4 mm Hg), diastolic (20 +/- 6 vs 12 +/- 3 mm Hg), and mean (23 +/- 7 vs 15 +/- 3 mm Hg) PA pressure; PA/systemic pressure ratio for systolic (0.37 +/- 0.08 vs 0.25 +/- 0.06) and mean (0.56 +/- 0.19 vs 0.29 +/- 0.11) pressures; and pulmonary vascular resistance (PVR) (5.63 +/- 1.06 vs 3.53 +/- 0.75 U). Aqueous oxygen (AO) infusion significantly reduced (p < 0.05) the values obtained with hypoxic ventilation; systolic (23 +/- 5 vs 30 +/- 7 mm Hg), diastolic (11 +/- 4 vs 20 +/- 6 mm Hg), and mean (14 +/- 3 vs 23 +/- 7 mm Hg) PA pressure; PA/systemic pressure ratio for systolic (0.25 +/- 0.05 vs 0.37 +/- 0.08) and mean pressures (0.29 +/- 0.12 vs 0.56 +/- 0.19); and PVR (3.41 +/- 1.01 vs 5.63 +/- 1.06 U). AO infusion in the pulmonary circulation did not influence the myocardial O2 atrioventricular (AV) difference or the O2 extraction. CONCLUSIONS: Infusion of hyperbaric AO solution into the PA can completely reverse the negative effects of acute hypoxia on the pulmonary circulation without affecting the myocardial metabolism. PMID- 15337028 TI - Myocardial protection with postconditioning is not enhanced by ischemic preconditioning. AB - BACKGROUND: Ischemic preconditioning (IPC) has been used in off-pump coronary artery bypass surgery (OPCAB) to reduce potential injury secondary to ligation of the target vessel. Previous studies have shown that a brief period of repetitive coronary occlusion applied at the onset of reperfusion, postconditioning (postcon), attenuates myocardial injury. This study tested the hypothesis that coincident application of IPC and postcon would provide more cardioprotection than either intervention alone by inhibiting oxidant-mediated injury after ischemia and reperfusion. METHODS: Four groups of open-chest canines endured 60 minutes coronary occlusion followed by 3 hours reperfusion: control (n = 10), no intervention; IPC (n = 9), 5 minutes left anterior descending coronary artery occlusion preceded 10 minutes of reperfusion before prolonged occlusion; postcon (n = 10), 3 cycles of 30 seconds reperfusion-30 seconds reocclusion were imposed immediately upon reperfusion; IPC+postcon (n = 8), IPC and postcon algorithms were combined. RESULTS: Collateral blood flow during ischemia was similar in all groups. Compared to control (24% +/- 2%), infarct size was comparably reduced in IPC (13% +/- 2%* [* denotes p less than 0.05 compared with control]), and postcon (10% +/- 1%*), consistent with a reduction in plasma creative kinase activity in these groups; infarct size was not further reduced by IPC+postcon (12% +/- 3%*). Tissue water content in ischemic myocardium was comparably reduced in IPC, postcon, and IPC+postcon compared to control. Superoxide anion generation detected by dihydroethidium staining in area at risk myocardium was comparably reduced in all intervention groups relative to control. Plasma malondialdehyde (microM), a lipid peroxidation byproduct of oxidant injury, was less at 1 hour of reperfusion in IPC (2.2 +/- 0.2*), postcon (2.1 +/- 0.2*), and IPC+postcon (2.5 +/- 0.2*) relative to control (3.3 +/- 0.2). Ventricular fibrillation occurred less often in all intervention groups. CONCLUSIONS: No additive cardioprotective effects by IPC and postcon were observed in a canine model of regional ischemia and reperfusion. The potent attenuation of myocardial injury by postcon may suggest a clinically applicable strategy during some surgical revascularization procedures (ie, OPCAB). PMID- 15337030 TI - Pinacidil improves contractile function and intracellular calcium handling in isolated cardiac myocytes exposed to simulated cardioplegic arrest. AB - BACKGROUND: We examined the effects of pinacidil on contractile function and intracellular calcium in isolated rat cardiomyocytes exposed to cardioplegic solution. METHODS: Rat myocytes were incubated at 24 degrees C for 2 hours in cardioplegic solution with or without pinacidil (50 micromol/L), then they were perfused with Krebs-Henseleit solution with a gas phase of 95% O2/5% CO2 at the same temperature. Contraction and intracellular calcium transients were then measured by video tracking and spectrofluorometry. RESULTS: During 20 minutes of perfusion after 2 hours in cardioplegic solution with pinacidil, (1) the recovery of contractile function was significantly increased in terms of both amplitude of contraction (98.30% +/- 9.90% versus 81.00% +/- 11.25%; p < 0.05) and peak velocity of cell shortening (100.90% +/- 13.79% versus 76.89% +/- 18.14%; p < 0.01) when compared with myocytes in cardioplegic solution without pinacidil; (2) the amplitudes of the intracellular calcium transients evoked by electrical stimulation and caffeine (10 mmol/L) increased by 23.31% to approximately 40.72% and 61.73%, respectively, compared with those in cardioplegic solution without pinacidil; and (3) the decay time of the caffeine-induced intracellular calcium transient decreased by 36.64% +/- 15.10% relative to that measured in cardioplegic solution without pinacidil. The effects induced by supplementing the cardioplegic solution with pinacidil were diminished in the presence of glibenclamide (10 micromol/L). CONCLUSIONS: Addition of the adenosine triphosphate-sensitive potassium-channel opener, pinacidil, to a high potassium cardioplegic solution improves recovery of contractile properties and cytosolic calcium in isolated rat cardiac myocytes. PMID- 15337031 TI - Outcomes after resection of giant emphysematous bullae. AB - BACKGROUND: Giant emphysematous bullae represent a rare form of emphysematous lung destruction. Surgical resection has traditionally been indicated when there is hyperexpansion of the chest, compromised pulmonary function, and evidence of underlying, relatively normal compressed lung. We review our experience and intermediate-term follow-up after the resection of giant bullae. METHODS: Forty three patients underwent resection of giant emphysematous bullae at Barnes-Jewish Hospital between March 1994 and June 2002. All had limiting dyspnea and radiologic evidence of hyperinflated giant bullae compressing adjacent lung parenchyma. Forty-one patients underwent preoperative pulmonary rehabilitation. Twenty-two patients underwent a bilateral procedure and 21 underwent a unilateral procedure. Mean follow-up was 4.5 years. RESULTS: One early death occurred on postoperative day 20 from heparin-induced thrombocytopenia and pulmonary embolism. Complications included prolonged air leak of more than 7 days in 23 (53%), atrial fibrillation in 5 (12%), postoperative mechanical ventilation in 4 (9%), and pneumonia in 2 (5%). Kaplan-Meier survival at 1, 3, and 5 years was 98%, 92%, and 89%, respectively. Four late deaths occurred at 1.4, 2.8, 3.5, and 5.9 years. Functional measures preoperatively and at 6 months and 3 years postoperatively were a forced expiratory volume in 1 second L (% predicted) of 1.2 +/- 0.6 (34%), 1.9 +/- 0.9 (55%), and 1.5 +/- 0.8 (49%); residual volume L (% predicted) of 5.1 +/- 1.2 (262%), 3.6 +/- 1.2 (154%), and 4.1 +/- 2.2 (209%); 6 minutes walk (ft) of 1230 +/- 361, 1393 +/- 300, and 1271 +/- 423; supplemental O2 used continuously (% patients) of 42%, 9%, and 21%; and O2 used during exercise of 73%, 37%, and 42%, respectively. CONCLUSIONS: In a contemporary series, giant bullectomy is shown to produce significant immediate functional improvement. This benefit declines with time but persists at least 3 years. PMID- 15337032 TI - Sports-related spontaneous pneumomediastinum. AB - BACKGROUND: Spontaneous pneumomediastinum is a rare medical entity occurring almost exclusively in otherwise healthy young individuals without known predisposing factors. We reported our experience with patients presenting with spontaneous pneumomediastinum related to sports. METHODS: Between January 1991 and December 2002, 10 patients were admitted with spontaneous pneumomediastinum related to sporting activities. We retrospectively reviewed their medical records with regard to predisposing factors, clinical presentation, diagnostic evaluation, and outcome. RESULTS: The mean age of our patients was 18.9 years (range 15 to 25 years). Retrosternal chest pain was the most common symptom (90%), and subcutaneous emphysema the most common physical finding (90%). Hamman's sign was present in 9 patients. In all 10 patients, physical exertion during sports was the only implicating factor (scuba diving in 4 patients, basketball in 2, soccer in 3, and volleyball in 1). All patients were treated conservatively. Complete resorption of the pneumomediastinum occurred in 3 to 8 days. Hospital stay ranged from 2 to 6 days (mean 3.8 days). Follow-up was complete in 8 patients (80%), and ranged from 6 to 84 months (mean 35 months). Only 1 recurrence of pneumomediastinum was found that was treated similarly. CONCLUSIONS: Spontaneous pneumomediastinum after sporting activities is a benign disease that generally resolves without clinical sequelae. Restriction of physical and athletic activity after the first episode is unnecessary, the only exception being that of scuba diving. PMID- 15337033 TI - Video-assisted thoracoscopic surgery of mediastinal bronchogenic cysts in adults: a single-center experience. AB - BACKGROUND: Mediastinal bronchogenic cysts are rarely diagnosed in adults, hence surgical experience is limited particularly with regard to video-assisted thoracoscopic surgery. In support of the thoracoscopic approach we report our single-center experience in this rare entity. METHODS: Between June 1995 and December 2002, a nonselected series of 12 consecutive patients presenting with mediastinal bronchogenic cysts underwent video-assisted thoracoscopic surgery. Six cysts (50%) had been diagnosed 2 to 22 years prior, only three of which became symptomatic. In asymptomatic patients (n = 7) surgery was performed because of increasing cyst size (n = 3), patient's request (n = 3), or suspected metastasis (n = 1). RESULTS: Mediastinal bronchogenic cysts were correctly diagnosed by computed tomography in 83% (10/12) and by magnetic resonance imaging in 100% (9/9). Using a three-trocar technique thoracoscopic surgery was successfully performed in 11 of 12 cases (92%). We noted no signs of acute cyst infection. No serious postoperative complications were observed. In 1 patient conversion to open thoracotomy was necessary due to extensive pleural adhesions. In another case thoracoscopic excision of the cyst wall was incomplete. Patients with thoracoscopic excision were discharged after a median of 5.5 days (range 4 to 14 days). No recurrences or complications were observed during a mean follow up of 40.5 months. CONCLUSIONS: Considering the low conversion and complication rate in our series, video-assisted thoracoscopic surgery should be the primary therapeutic choice among adults with symptomatic mediastinal bronchogenic cysts. Surgical intervention in patients with asymptomatic and uncomplicated cysts appears optional. PMID- 15337034 TI - Prognosis of thymic epithelial tumors according to the new World Health Organization histologic classification. AB - BACKGROUND: The aim of this study was to document the prognosis of thymic epithelial tumors (TETs) according to new the World Health Organization (WHO) classification. METHODS: We retrospectively reviewed 150 patients with TETs that were confirmed pathologically during 11 years (from 1992 to 2002) in Severance Hospital, Seoul, Korea. RESULTS: TETs were classified as type A, AB, B1, B2, B3, or C, tumors and these represented 7 (4.7%), 26 (17.3%), 13 (8.7%), 45 (30.0%), 26 (17.3%), and 33 (22.0%) cases, and the 5-year survival rates were 100%, 93%, 89%, 82%, 71%, and 23%, respectively. Their Masaoka stages were I, II, III, IVa, and IVb, with 53 (35.3%), 39 (26.0%), 20 (13.3%), 22 (14.7%), and 16 (10.7%) cases. Tumor invasiveness, recurrence, completeness of resection, and tumor related death were more frequent in types AB, B2, B3, and C than in types A and B1. Multivariate analysis showed that Masaoka stage (p < 0.001) and the WHO classification (p = 0.019) were significant independent prognostic factors. CONCLUSIONS: The WHO classification is associated with tumor invasiveness, recurrence, completeness of resection, and tumor-related death, and has good correlation with Masaoka stage. The WHO histologic subtypes are an independently significant prognostic factor with respect to survival in our multivariate analysis. Types AB, B2, B3, and C showed invasive behaviors and R1 or R2 resections were frequently performed. Postoperative adjuvant radiotherapy was effective, but long-term follow-up is recommended because of decreased survival after 5 years following operation. The WHO classification may be helpful in clinical practice for the assessment and treatment of TET patients. PMID- 15337036 TI - Recent results of postoperative mortality for surgical resections in lung cancer. AB - BACKGROUND: Changes in the postoperative mortality rates and causes of death for lung cancer surgery at the specialized hospital for cancer in Tokyo, Japan during the last 16 years were investigated. METHODS: Data on 3,270 consecutive patients who underwent pulmonary resection for primary lung cancer between January 1987 and December 2002 at the National Cancer Center Hospital were retrospectively analyzed. The postoperative 30-day and in-hospital mortality rates and causes of death after pulmonary resection for lung cancer were investigated. Patients were divided into two period groups of almost equal number, the early (1,615 patients from 1987 to 1996) and the late (1,655 patients from 1997 to 2002) periods. RESULTS: Fifty-eight operative and postoperative deaths occurred during the last 16 years. Thirty-day and in-hospital mortality were 0.6% (21/3,270) and 1.6% (58/3,270), respectively. During the last 6-year period, 30-day and in-hospital mortality were 0.5% (8/1,655) and 0.8% (21/1,655), respectively. The difference was significant between the 30-day/in-hospital mortality for pneumonectomy (3.1%/5.9%) and lobectomy (0.3%/1.3%) (p < 0.0001/p < 0.0001). The difference in mortality between lobectomy and segmentectomy or a lesser resection was not significant. The 58 deaths were caused by pneumonia/acute respiratory distress syndrome (ARDS) (36%, n = 21), bronchopleural fistula (BPF)/empyema (33%, n = 19), cerebrovascular accident (10%, n = 6), cardiac-related event (7%, n = 4), and others (14%, n = 8). The most frequent cause of death in the early period was BPF/empyema (18/45, 40%), while that in the late period was pneumonia/ARDS (6/13, 46%). Among the pneumonia/ARDS deaths in the late period (n = 6), 5 (83%) were due to acute deterioration of interstitial lung disease after lobectomy. CONCLUSIONS: Recent postoperative mortality rates (30-day, 0.5%; in-hospital, 0.8%) in the treatment of lung cancer are quite acceptable. Special care must be taken for the patient after pneumonectomy, as reported by others. Furthermore, even after lobectomy, proper management of the patient with acute deterioration of interstitial lung disease will be required to improve the future outcome. PMID- 15337038 TI - Effect of histologic type and smoking status on interpretation of serum carcinoembryonic antigen value in non-small cell lung carcinoma. AB - BACKGROUND: Serum carcinoembryonic antigen (CEA) has all of the properties desired for a biologic measure to be used as a prognostic indicator in the clinical evaluation of lung cancer. Carcinoembryonic antigen value appears to be related to tumor histologic type and patients' smoking status, which has yet to be intensively analyzed as reports available thus far have consisted of a limited number of patients. This study was undertaken to determine whether the prognostic value of CEA differs according to histologic type in a large group of patients with clinical early-stage lung cancer, and how smoking influences its value. METHODS: Two series of 694 and 260 consecutive patients who underwent resection for clinical stage I lung adenocarcinoma and squamous cell carcinoma, respectively, were evaluated. We measured serum CEA before and after surgery, and analyzed its prognostic significance in relation to histologic type and its correlation with smoking status. RESULTS: We found significantly higher CEA levels in patients with adenocarcinomas than in those with squamous cell carcinomas (7.8 versus 5.5 ng/mL; p = 0.0018), but a higher percentage of CEA positive patients among those with squamous cell carcinoma (109 of 260, 41.9%) than those with adenocarcinoma (245 of 694, 35.3%). Clinical stage I patients with a high preoperative CEA level had a poor prognosis, and for pathologically confirmed stage I patients with a high postoperative CEA level the prognosis was worse. The prognostic value of serum CEA level was thus significantly greater for adenocarcinoma than for squamous cell carcinoma. This was probably because of a much higher proportion of smokers among patients with squamous cell carcinoma. In adenocarcinoma, the growth of which was generally less influenced by smoking, the proportion of CEA-positive smokers (49.3%, 170 of 345) was greater than that of CEA-positive nonsmokers (21.5%, 75 of 349, p < 0.0001). Additionally, in patients with adenocarcinoma, survival of nonsmokers was more greatly influenced by CEA level than that of smokers. CONCLUSIONS: Although serum CEA values measured before and after surgery are important in identifying patients at high risk of poor survival, its specificity is higher for adenocarcinoma than for squamous cell carcinoma. When serum CEA levels are checked, smoking status of patients, particularly of those with squamous cell carcinoma, should be taken into account. PMID- 15337040 TI - A clinicopathological study of resected adenocarcinoma 2 cm or less in diameter. AB - BACKGROUND: The biological behavior of small adenocarcinoma is different in each patient and these are especially enormous differences when evaluating solid tumors and nonsolid tumors. METHODS: A total of 159 adenocarcinomas 2 cm or less in diameter were studied. Several clinicopathological factors were retrospectively analyzed. RESULTS: The diameter of the primary tumors was less than 1 cm in 47 patients, 1-1.5 cm in 49 patients, and 1.5-2 cm in 63 patients, respectively. Almost all patients (147) were pathologic N0 and there were 12 node positive patients (7.5%). Lymph-node involvement was observed in 1 patient with a tumor diameter measuring less than 1 cm and in 11 patients with a tumor diameter measuring 1-2 cm. According to Noguchi' s classification, 33 patients belonged to class A or B, 71 patients belonged to class C, and 55 patients belonged to class D, E, or F. The ratio of ground-glass opacity (GGO) area in the main tumor in high resolution computed tomography was classified into two groups with a threshold of 50%. There were 44 patients with a GGO ratio of equal to or greater than 50%, none of which indicated lymph-node metastasis or tumor recurrence during follow-up (5-year survival = 100%). On the contrary among 115 patients with a GGO ratio less than 50%, lymph-node involvement was indicated in 12 patients (10.4%) and the 5-year survival rate was 83.9%. CONCLUSIONS: The biological malignancy of small adenocarcinomas might be accurately evaluated by the proportion of GGO area as well as the Noguchi classification. PMID- 15337041 TI - The accuracy of integrated PET-CT compared with dedicated PET alone for the staging of patients with nonsmall cell lung cancer. AB - BACKGROUND: The treatment of patients with nonsmall cell lung cancer (NSCLC) is determined by the stage. We evaluated the accuracy of staging using integrated positron emission tomography (PET) and computed tomography (CT) and compared it with dedicated PET visually correlated with CT scan. METHODS: A prospective blinded trial was performed on a consecutive series of patients with NSCLC. Patients underwent integrated PET-CT scanning with 2-[18F]-fluoro-2-deoxy-D glucose (FDG-18). A radiologist assigned the T, N and M status. No sooner than 2 weeks the same radiologist read the dedicated PET alone, without the integrated CT images and a T, N and M status was assigned again. The most recent CT scan was available and visually correlated with both studies. All patients underwent biopsies of suspicious N2 or N3 lymph node or distant metastases and if negative, pulmonary resection with lymphadenectomy was performed. RESULTS: There were 129 patients. Integrated PET-CT is a better predictor than PET for all stages of cancer and achieved statistical significance for stage I (52% versus 33%, p = 0.03) and for stage II (70% versus 36%, p = 0.04). It also is a better overall predictor for T status (70% versus 47%, p = 0.001) and the N status (78% versus 56%, p = 0.008). Nodal analysis shows that integrated PET-CT was more accurate for the total N2 nodes (96% versus 93%, p = 0.01) and for the total N1 nodes (90% versus 80%, p = 0.001). It was also more sensitive, specific, and had a higher positive predictive value for both N2 and N1 nodes (p < 0.05 for all). Integrated PET-CT is significantly more sensitive at the 4R, 5, 7, 10 L and 11 stations and more accurate at the 7 and 11 lymph nodes stations than dedicated PET. CONCLUSIONS: Integrated PET-CT using FDG-18 better predicts stage I and II disease as well as the T and N status of patients with NSCLC when compared with dedicated PET alone. It is more accurate at some nodal stations but still only achieves an accuracy of 96% and 90% for the N2 and N1 nodes, respectively. PMID- 15337042 TI - Surgical management of thoracic malignancies invading the heart or great vessels. AB - BACKGROUND: Surgical resection of thoracic malignancies involving either the heart or great vessels is uncommonly performed because of the potential morbidity and mortality for an unknown probability of significant palliation or cure. We reviewed our experience of 10 patients treated surgically, either primarily or as a component of multimodality therapy, to assess feasibility and results. METHODS: A retrospective review of the results in 10 patients who underwent resection of thoracic malignancies that included either great vessel or the heart was conducted. RESULTS: Histologic diagnoses included soft tissue sarcoma (n = 7), squamous cell carcinoma (n = 1), malignant thymoma (n = 1), and mediastinal teratoma (n = 1). Three patients underwent induction chemotherapy. Cardiopulmonary bypass was used in 7 patients. Structures resected included superior vena cava (n = 5), left atrium (n = 4), right atrium (n = 2), descending aorta (n = 1), and main pulmonary artery (n = 1). Concomitant anatomic pulmonary resections were performed in 3 patients. Seven patients had an R0 or R1 resection. There were no perioperative deaths. All symptomatic patients had immediate and sustained palliation of their presenting symptoms. The median length of stay was 6 days (range, 4 to 43 days). Six patients underwent postoperative systemic therapy. The overall median survival was 21.7 months (range, 3.2 to 69 months) and was 33.3 months (range, 3.7 to 69 months) for patients who had an R0 or R1 resection. CONCLUSIONS: Resection of the heart and great vessels involved by thoracic malignancies can be performed with acceptable morbidity and mortality and results in significant palliation and, in some cases, prolonged survival. PMID- 15337043 TI - Thoracoscopy using a substernal handport for palpation. AB - BACKGROUND: A substernal handport allows palpation of the lung and thus circumvents one of the major limitations of thoracoscopy. METHODS: This approach has been used in 24 consecutive patients, primarily during planned metastasectomy or when palpation was needed for deeper or smaller lesions that were difficult to find. RESULTS: No long-term complications from this procedure were noted, and the 3 early complications were either minor or unrelated to the procedure. This approach allowed adequate resection to be accomplished by a less invasive approach in 67% of patients, although conversion to an open procedure was necessary in 33% of patients for anatomic and technical reasons. Among the 16 patients who underwent this procedure alone, the median length of stay in the hospital was 3 days. The rate of incomplete resection and of recurrence after metastasectomy was comparable to that for an open approach. CONCLUSIONS: Our experience documents that a substernal handport is safe, does not compromise the ability to perform an adequate metastasectomy, and allows biopsy of lesions that are otherwise not amenable to a minimally invasive approach. This technique should be included in the standard armamentarium of approaches for thoracic surgery. PMID- 15337044 TI - Incidence of anemia in patients receiving neoadjuvant chemotherapy for locally advanced esophagogastric cancer. AB - BACKGROUND: There is rising evidence that anemia and blood transfusion increase perioperative mortality in cancer patients. Patients who are treated with neoadjuvant chemotherapy with a curative intent are exposed to toxicity that may negatively affect their future outcome. METHODS: The charts of 29 patients (21 males; median age, 59.5 years; range, 37 to 73), receiving neoadjuvant chemotherapy for cT3 esophagogastric adenocarcinoma operated at a single university center in the year 2002, were retrospectively reviewed to assess the incidence of anemia and blood transfusions. RESULTS: Twenty-six patients received platinum-based chemotherapy over a period of 12 weeks and three patients more than 6 weeks. The median hemoglobin level (Hb level) before chemotherapy was 14.0 g/dL (range, 10.4 to 15.9 g/dL), the median decline of the Hb level was 2.9 g/dL (range, 0.3 to 6.3 g/dL); this drop was statistically significant (p < 0.001, 95% confidence interval). Patients who received preoperative blood transfusions (n = 8, 28%) had a significantly increased risk of developing postoperative complications (p = 0.028). CONCLUSIONS: Preoperative chemotherapy for locally advanced esophagogastric cancer induces anemia and therefore leads to preoperative blood supplementation in a considerable number of patients. Data indicate that this may counteract the beneficial effects of neoadjuvant treatment. PMID- 15337045 TI - Effect of inhibition of multiple steps of angiogenesis in syngeneic murine pleural mesothelioma. AB - BACKGROUND: Angiogenesis is a multistep process in which the endothelial cell plays a pivotal role. We hypothesized that the combination of two antiangiogenic agents with distinct mechanisms of action would more effectively inhibit tumor growth than either agent alone in a murine mesothelioma model. METHODS: A syngeneic murine mesothelioma flank tumor model (AB-12) was established in BALB/c mice. Separate adenovirus vectors expressing the cDNAs for human pigment epithelium-derived factor (AdPEDF) and a soluble form of the human vascular endothelial growth factor receptor-1 (Adsflt-1) were administered intratumorally. End points measured included tumor size, animal survival, and microvessel density using CD31 immunohistochemistry. An orthotopic model of mesothelioma was established by implanting AB-12 cells into the murine pleural cavity. Simultaneously, AdPEDF and Adsflt-1 were instilled intrapleurally and tumor burden and survival were recorded. The development of pulmonary emphysema was also assessed by calculating the mean linear intercept (a measure of interalveolar septal distance) in histologic lung sections from tumor-free mice after vector administration. RESULTS: In the flank tumor model, the combination of AdPEDF and Adsflt-1 inhibited tumor growth, prolonged survival, and decreased microvessel density more profoundly compared with either AdPEDF or Adsflt-1 alone. In the orthotopic model, the combination was also more effective in prolonging survival. Intrapleural AdPEDF or Adsflt-1 did not increase the mean linear intercept compared with controls in tumor-free mice. CONCLUSIONS: In this murine model, inhibiting multiple mechanisms of angiogenesis using two agents is a more effective antineoplastic strategy than using either agent alone. In addition, instillation of antiangiogenic gene transfer vectors into the pleural space does not result in histologic evidence of pulmonary emphysema. PMID- 15337046 TI - Selective division of T3 rami communicantes (T3 ramicotomy) in the treatment of palmar hyperhidrosis. AB - BACKGROUND: Compensatory sweating (CS) is the main cause of a patient's dissatisfaction after sympathetic surgery for palmar hyperhidrosis. Preservation of the sympathetic nerve trunk and limitations on the range of dissection are necessary to reduce CS. METHODS: We compared 64 patients (31 male, 33 female) (group 1) who underwent a T2 sympathicotomy between July 1998 and February 1999 and 83 patients (58 male, 25 female) (group 2) who underwent a T3 ramicotomy between August 2000 and December 2002. RESULTS: In group 1, 60 patients (93.8%) exhibited a decreased sweating on both hands, but 4 patients (6.2%) exhibited a persistent sweating on both hands. For group 2, 58 patients (69.9%) experienced a decreased sweating on both hands, 15 patients (18.1%) experienced a persistent sweating on both hands, and 10 patients (12.0%) experienced a persistent sweating on one hand. The grade of CS in group 2 was significantly lower than in group 1 (p < 0.001) and, notably, the rate of embarrassing and disabling CS in group 2 (15.5% [9 out of 58]) was significantly lower than in group 1 (43.3% [26 out of 60], p value < 0.001). The rate of satisfaction was 78.1% (50 out of 64) for group 1 and 68.6% (57 out of 83) for group 2 with no significant statistical difference indicated (p = 0.202). CONCLUSIONS: The incidence of sweating postoperatively was relatively high in the T3 ramicotomy group, although the T3 surgery did result in a lower incidence of CS when compared with a T2 sympathicotomy. PMID- 15337047 TI - Epicardial pulmonary vein isolation with a hook-shaped cryoprobe to treat atrial fibrillation. AB - PURPOSE: After it was shown that a rapidly firing focus in a pulmonary vein (PV) can cause atrial fibrillation, percutaneous endocardial PV isolation using radiofrequency began to be used as a method of treatment. However, this technique is time consuming. It requires fluoroscopy and contrast media to identify the PV, and cardiac tamponade and PV obstruction are major complications. To overcome these drawbacks, we developed a hook-shaped cryoprobe to enable circumferential ablation of PV orifices epicardially. The aim of this experimental study in dogs was to confirm the efficacy of this method electrophysiologically. DESCRIPTION: Five mongrel dogs (32 PVs) were used. Surrounding tissue was dissected to expose all PVs and their orifices into the left atrium. Each PV was stimulated with an electrode to measure the length of PV which has the same pacing threshold as the left atrium. EVALUATION: The mean distance from the PV orifice to the pacing boundary line was 8.9 +/- 1.3 mm. Encircling cryoablation was performed with a hook-shaped cryoprobe to circumferentially ablate each PV orifice epicardially. CONCLUSIONS: Cryoablation by this method created a bidirectional conduction block in all PVs. All PVs were electrically isolated, and the PV isolation was achieved epicardially without atriotomy. This method should enable less invasive treatment of AF clinically. PMID- 15337048 TI - A novel, form-stable, anatomically curved vascular prosthesis for replacement of the thoracic aorta. AB - PURPOSE: Current replacement of the thoracic aorta performed with straight vascular prostheses may cause kinking, potentially affecting hemodynamics and promoting vortices and thrombus formation. A novel vascular prosthesis, resistant to pressure-related shape deformation, was designed to imitate the curved anatomy of the thoracic aorta. DESCRIPTION: A woven velour prosthesis was trimmed with cross-sutures along a marked line, resulting in a curved-shaped anatomic form, and was compared with conventional straight and thermally fixed curved grafts. The vascular prostheses were fixed at both ends at various base distances (8, 10, 12, 14, and 16 cm) and pressurized. To imitate the neck vessels an abutment was fixed at the upper convexity of the grafts. Radius of curvature or depth of kinking was measured at different pressures (100, 125, and 150 mm Hg). Pressure gradients and flow profiles were further analyzed in an aortic arch glass model. EVALUATION: When pressurized the straight and the thermally fixed protheses showed double kinking before and behind the abutment at all pressures and distances. Kinking depth increased with increasing pressure and less base distance. Transkinking pressure gradients increased with the degree of kinking. In a glass model flow profiles showed postkinking turbulences and vortex formation. The newly designed vascular prosthesis showed no kinking and remained form stable at all test conditions. CONCLUSIONS: This novel curved vascular prosthesis for replacement of the thoracic aorta demonstrates form stability compared with conventional straight and thermally fixed vascular prostheses in an aortic arch model, with smaller pressure gradients and flow disturbances. PMID- 15337050 TI - Morphological and functional magnetic resonance imaging after heterotopic heart transplantation. AB - In patients with severe congestive heart failure, a marked elevation in pulmonary vascular resistance limits the success of orthotopic cardiac transplantation, providing the rationale for heterotopic transplantation. In the case reported, cardiac anatomy and function of two hearts in the same chest were imaged using magnetic resonance imaging (MRI). MRI offers a high-resolution, three dimensional, noninvasive technique to visualize the complex anatomy after heterotopic heart transplantation, providing information of morphologic and functional parameters at the same time. The challenge of sufficient electrocardiogram triggering, hindered by two hearts with electrophysiological activity in the same chest, can be overcome using new real-time techniques. PMID- 15337051 TI - Bidirectional cavopulmonary shunt for acute right ventricular failure in an adult patient. AB - Right ventricular failure due to right coronary occlusion is an uncommon but serious complication of acute aortic dissection. We report a patient with right coronary occlusion from acute type A aortic dissection who developed right ventricular failure that persisted after proximal aortic repair, rendering weaning from extracorporeal circulation unsuccessful. With a bidirectional cavopulmonary shunt and the use of an intraaortic balloon, the patient could be weaned successfully from cardiopulmonary bypass. This procedure may be an alternative to a right ventricular assist device in selected candidates. PMID- 15337052 TI - Pseudoaneurysm of homograft placed in right ventricular outflow tract. AB - Pseudoaneurysm of the right ventricular outflow tract after homograft placement is an infrequent complication after intracardiac repair for tetralogy of Fallot. We report two cases of pseudoaneurysm of right ventricular outflow tract after homograft placement for surgical repair of tetralogy of Fallot with pulmonary atresia. PMID- 15337053 TI - Diagnostic criteria for penetrating atheromatous ulcer of the thoracic aorta. AB - An autopsy case of penetrating atheromatous ulcer of the ascending thoracic aorta is presented. The triplet imaging findings of mediastinal dilatation, interstitial pulmonary hemorrhage and pericardial effusion were diagnostic for penetrating atheromatous ulcer at the posterior wall of the ascending thoracic aorta. Immediate surgical treatment is indicated when these three findings emerge during examination. PMID- 15337054 TI - Stent-graft placement combined with percutaneous fenestration for the treatment of aortic dissection in a patient at risk of developing renal ischemia. AB - A 51-year-old man, suffering from an aortic dissection with the entry tear in the descending thoracic aorta, underwent replacement of the ascending aorta and aortic arch with resection of the entry tear. Although the false lumen of the descending thoracic aorta was thrombosed after surgery, redissection developed in the descending aorta 2 months later. Percutaneous fenestration was performed before stent-grafting because it was judged that there was a significant risk of left renal ischemia. Entry closure with a stent-graft was performed successfully and no renal ischemia developed. The patient is doing well at 1 year after the procedure. PMID- 15337055 TI - Open aortic surgical repair for left hemi-arch stent-graft failure. AB - A surgical technique of endovascular graft explant through an open aortic approach for left hemi-arch stent-graft failure is described. Between January and April 2003, we surgically treated 3 patients previously submitted for stent grafts for isthmic aortic diseases. Two patients had atherosclerotic aneurysm and 1 had a false lumen reperfusion of subacute intramural hematoma. At 6 to 8 months computed tomographic scan follow-ups on all patients showed a rapid enlargement of aortic diameters due to type I endoleaks. The presence of an uncovered proximal stent in the parasubclavian aorta did not allow a simple aortic cross clamping; therefore we performed an open aortic procedure through a left posterolateral thoracotomy, using femoro-femoral bypass and mild hypothermic circulatory arrest. Selective antegrade cerebral perfusion was started within 3 to 5 minutes from aortotomy and graft removal. Left hemi-arch and descending thoracic aortic replacement was then performed with continuous cerebral perfusion. No surgical mortality was observed. Postoperative course was uneventful for neurologic, cardiac, respiratory, and renal complications. The 3 month follow-ups were event free. This approach, associated with rapid stent graft explant and selective cerebral antegrade perfusion, appears to be a safe and effective surgical strategy for treating this new aortic pathology. PMID- 15337056 TI - Coronary-coronary radial artery graft for single, distal LAD lesion. AB - Radial artery segment has been used as a coronary-coronary graft for single, distal lesion of the large left anterior descending coronary artery running well over the cardiac apex. In our opinion this technique may occasionally be an attractive approach for bypassing very distal lesions of large coronary arteries combined with regular arterial or venous grafting of the target artery if proximal stenosis is also present. The remnant of the radial artery can be used for grafting of another diseased artery (eg, the large first marginal branch of the circumflex artery in our case report). PMID- 15337057 TI - Application of Cabrol technique to off-pump coronary artery bypass grafting using radial artery. AB - We present the case of a Cabrol-type proximal anastomosis technique in off-pump coronary artery bypass. The patient was a 64-year-old man with significant stenoses on the left main, left anterior descending, and proximal right coronary artery. The obtuse marginal and right coronary arteries were anastomosed with both ends of a radial artery. For the proximal anastomosis during this procedure, we applied the Cabrol-type looping interposition technique. In selected patients, we suggest that this technique allows the effective use of graft length and can reduce a number of ascending aortic manipulations. PMID- 15337058 TI - Anomalous origin of the left coronary artery from the pulmonary artery in an adult with systemic collateral circulation to the left coronary artery. AB - A case of anomalous origin of the left coronary artery from the pulmonary artery in an adult with collateral circulation between the left coronary artery and systemic extracardiac vessels is reported. After evaluating the surgical options, my colleagues and I conclude that ligation of the left coronary artery and a left internal thoracic artery graft to the left anterior descending coronary artery is the preferable option for treating such patients. PMID- 15337059 TI - AIDS-related cardiac tamponade: is surgical drainage justified? AB - In order to evaluate the usefulness of surgical drainage in the treatment of patients with acquired immunodeficiency syndrome (AIDS)-related cardiac tamponade, we reviewed our experience with subxiphoid pericardiostomy on 5 consequent such patients. One patient died in the immediate postoperative period and the remaining 4 died within 21 weeks after the operation. Similar results have been reported by other authors who found that surgical drainage has no diagnostic or therapeutic benefit over pericardiocentesis in this particular group of patients. Based on our limited experience and the data of the literature, we feel that surgical drainage cannot be justified as the primary method of treatment of AIDS-related cardiac tamponade. PMID- 15337060 TI - Pericarditis as complication of appendicitis. AB - Pericarditis as a complication of appendicitis is a rare event. In a 25-year period we encountered two pediatric cases with this severe complication due to (a)typical presentation of appendicitis resulting in small bowel obstruction, intraabdominal abscesses, constrictive pericarditis, and purulent pericarditis. PMID- 15337061 TI - Partial hepatic vein diversion in pulmonary arteriovenous malformations in congenital heart disease. AB - The late appearance of pulmonary arteriovenous malformations after operations for complex congenital cardiac conditions may be explained in many ways. The necessity of hepatic blood flow to the lungs is now generally accepted for carrying some labile substance that is secreted by the liver and that has not yet been identified. Rerouting of hepatic blood to the lungs in cardiac patients can be accomplished with different methods, most of which are predisposed to thrombosis because of the slow flow in the tunnel. My colleagues and I describe a case of partial diversion of hepatic blood into the pulmonary circulation and suggest a strategy to decrease the likelihood of conduit thrombosis and to treat pulmonary arteriovenous malformations. PMID- 15337062 TI - Comparison of right ventricle to pulmonary artery conduit and modified Blalock Taussig shunt hemodynamics after the Norwood operation. AB - The Norwood procedure remains one of the highest-risk operations in congenital heart surgery. A significant contributor to this risk is thought to be the diastolic run-off into the modified Blalock-Taussig shunt (MBTS). In an effort to eliminate this risk, several groups have begun to utilize a right ventricle to pulmonary artery conduit (RVPAC), which decreases this diastolic "steal" of coronary blood flow. Whereas initial results with the RVPAC are encouraging, the postulated hemodynamic advantages are unproven. This case illustrates the positive hemodynamic changes by echocardiography after the replacement of a MBTS with a RVPAC in a patient after a Norwood procedure. PMID- 15337063 TI - Sleeve resection of a transcarinal bronchial carcinoid after laser debulking. AB - We report a case of a bronchial carcinoid tumor extending from the right upper lobe into the left mainstem bronchus in a 30-year-old woman. Diagnosis was established by preoperative bronchoscopy and biopsy. After extensive debulking with seven sessions of bronchoscopic neodymium:yttrium-aluminum-garnet laser therapy, the tumor was resected by right upper-lobe sleeve lobectomy. Final pathology revealed a typical carcinoid tumor with surgical margins and all lymph nodes free of tumor. PMID- 15337064 TI - Sudden cardiac herniation 6 months after right pneumonectomy. AB - Cardiac herniation is a rare complication of intrapericardial pneumonectomy and has a high mortality. The condition has been reported only within 24 hours after surgery. In this report, a case is described in which a total cardiac herniation took place 6 months after right intrapericardial pneumonectomy. The patient presented with an acute vena cava superior syndrome and underwent thoracotomy to reposition the heart into the pericardial sac and to close the pericardium with a patch. PMID- 15337065 TI - Endovascular stenting for aortic dissection with lower extremity malperfusion. PMID- 15337066 TI - Evaluation of a vertical vein using magnetic resonance imaging. PMID- 15337067 TI - Surgical anterior ventricular restoration in patients with aortic regurgitation. AB - My colleagues and I describe a novel method of perfusion during surgical anterior ventricular restoration in patients with aortic regurgitation. The modified technique was used in 2 patients with aortic incompetence who required the restoration operation. PMID- 15337068 TI - The mammary loop: how to do an adjustable "Y" graft with the left internal thoracic artery. AB - We present a technique that permits the grafting of two vessels with the left internal thoracic artery when a sequential graft cannot be performed. The left internal mammary artery is anastomosed to itself resulting in a loop that will be cut open at the time of the coronary anastomosis. PMID- 15337069 TI - A new method for graft tailoring in hemiarch replacement. AB - We describe a method of graft tailoring in hemiarch replacement that can accommodate long elliptical distal anastomosis and the right angle between the arch and ascending aorta without kinks, resulting in a more physiologic shape of the neoaorta. The distal end of the graft is beveled superiorly, to spare the origins of brachiocephalic vessels as in the standard bevel technique, but the incision line is curvilinear to form a concave cut end. Once the distal anastomosis is completed and the concavity of the cut end is straightened, the undersurface of the graft tongue forms a lesser curvature of the arch. PMID- 15337070 TI - A simple solution for management of the postpneumonectomy empyema cavity. AB - Postpneumonectomy bronchopleural fistula with empyema is a difficult problem. Once the fistula is healed, successful closure of the pleural space is associated with varying degrees of success, as well as the potential for major reconstructive surgery. My colleagues and I describe a simple approach for the definitive management of the open pleural space involving split-thickness skin grafts to marsupialize the pleural cavity. PMID- 15337071 TI - Apoptosis: pathophysiology and therapeutic implications for the cardiac surgeon. AB - Cardiomyocyte apoptosis has been associated with the pathogenesis of heart failure as well as ischemic and inflammatory myocardial conditions. The aim of this study is to give a critical synopsis of cardiomyocyte apoptosis and identify methods to prevent or attenuate apoptosis in patients undergoing cardiac surgery. Clinical conditions and agents associated with decreased apoptotic index are early repair or replacement of valvular pathology before deterioration of ventricular function, afterload reduction with medication or intraaortic balloon pulsation in patients with acute increase in afterload or in hemodynamically compromised patients, decreasing catecholamine-induced cardiotoxicity by using beta-blockers, phosphodiesterase inhibitors, or early insertion of intraaortic balloon pulsation or ventricular assist device. Prompt coronary revascularization, which reduces myocardial ischemia time, is the most effective antiapoptotic therapy. Reduction of myocardial apoptosis associated with cardiopulmonary bypass and aortic cross-clamping are other therapeutic targets. Some investigational therapies include ischemic preconditioning and use of antiapoptotic medication such as the caspase inhibitors, antioxidants, calcium channel blockers, the insulin-like growth factor-1, and the poly-adenosine diphosphate-ribose-synthetase inhibitors. Most of the therapeutic implications in reducing cardiomyocyte apoptosis are still in the experimental phase. Some options are already incorporated in the clinical practice of the cardiovascular surgeon. New therapeutic considerations include avoiding sustained and long-term use of catecholamines and reducing or avoiding cardiopulmonary bypass-when clinically feasible. Noncatecholamine inotropes should be preferred for patients undergoing heart failure surgery and for patients with low output syndrome after open-heart surgery. The lessons learned from apoptosis research reinforce more liberal and early insertion of intraaortic balloon pulsation or ventricular assist device in clinical low output states. PMID- 15337072 TI - Modification of bicaval anastomosis for orthotopic cardiac transplantation. PMID- 15337073 TI - Some technical considerations on the prevention of bleeding caused by sutures in the posterior pericardium in off-pump coronary artery bypass operations. PMID- 15337074 TI - Recommendations for the proper placement of the deep pericardial retraction suture in off-pump coronary artery bypass graft surgery. PMID- 15337075 TI - Aortic ball-valve prosthesis after 38 years. PMID- 15337076 TI - Benefits of off-pump bypass on neurologic function. PMID- 15337078 TI - Replacement of the trachea with an autologous aortic graft. PMID- 15337080 TI - The importance of obstructed and unobstructed segments to be resected in predicting postoperative FEV1. PMID- 15337081 TI - Pericardial synovial sarcoma: 14-year survival with multimodality therapy. AB - We report a case of recurrent primary synovial sarcoma of the pericardium. Reverse transcriptase-polymerase chain reaction analysis for t(X,18) demonstrated the presence of the chimeric transcript SYT/SSX. Because of the rarity of this entity, optimal therapy is unknown. The prognosis of this tumor is very poor in previous reports. In this report, we present a case with five recurrences treated by a combination of surgery, chemotherapy, and radiotherapy. The patient survives now for more than 14 years, the longest reported survival of a primary synovial sarcoma of the pericardium. PMID- 15337082 TI - Primary cardiac paraganglioma. AB - We report a case of a 29-year-old male, who during workup of hypertension was found to have a malignant primary paraganglioma of the heart. The tumor arose from the site of the aortopulmonary window and right ventricular outflow tract (RVOT) and was removed with the aid of cardiopulmonary bypass. Reconstruction of the RVOT and pulmonary valve was necessary because of involvement by the tumor. The surgical course was uncomplicated, with normalization of catecholamine secretion and blood pressure. PMID- 15337083 TI - Successful aortic root remodeling for repair of a dilated pulmonary autograft after a Ross-Konno procedure in early childhood. AB - We report a case of successful reoperation for pulmonary autograft root dilatation causing severe regurgitation after a Ross-Konno procedure in an infant. The procedure consisted of reduction of the circumference of the sinotubular junction and the autograft annulus. This technique is an effective alternative to prolong the transition period after a Ross procedure prior to other definitive surgeries, such as valve-sparing aortic root replacement or prosthetic valve replacement, which are preferably avoided at this young age. PMID- 15337084 TI - Aerosolized iloprost as a bridge to lung transplantation in a patient with cystic fibrosis and pulmonary hypertension. AB - We describe a patient with cystic fibrosis, end-stage lung disease, and secondary pulmonary hypertension in whom aerosolized iloprost was effective in lowering pulmonary artery pressure and improving functional status, thus proving successful as a bridge to lung transplantation. Inhaled iloprost may be an efficient and selective approach to treat pulmonary hypertension related to end stage obstructive pulmonary disease. PMID- 15337085 TI - Lymphangioma of the esophagus: surgical treatment. AB - Lymphangioma of the esophagus is a very rare submucosal tumor, but it can usually be removed without complication by endoscopic resection because of its small size and benign nature. My colleagues and I report a case of an uncommonly large lymphangioma (5.1 x 2.3 x 1.7 cm) in the distal esophagus treated by distal esophagectomy and esophagogastrostomy for complete resection without narrowing the esophagus. The patient's immediate postoperative course was uneventful, and he remains well 6 months after the operation. PMID- 15337086 TI - Compression of the left main bronchus and pulmonary artery after entire aortic replacement with Dacron arterial grafts. PMID- 15337087 TI - Persistent gastric stump-pleural fistula with circumscribed pleural empyema: a single approach for a twofold problem. PMID- 15337088 TI - Median sternotomy for reoperation of the distal aortic arch in postcoarctectomy patients. AB - We report the usefulness of a median sternotomy approach for the surgical treatment of residual aortic gradients or recoarctation and aortic aneurysms. This series confirms that excellent surgical results can be obtained in this technically challenging group of patients needing postcoarctation repair. PMID- 15337090 TI - WRKY transcription factors: from DNA binding towards biological function. AB - WRKY proteins comprise a large family of transcription factors. Despite their dramatic diversification in plants, WRKY genes seem to have originated in early eukaryotes. The cognate DNA-binding site of WRKY factors is well defined, but determining the roles of individual family members in regulating specific transcriptional programs during development or in response to environmental signals remains daunting. This review summarises the recent advances made in starting to unravel the various functions controlled by WRKY proteins. PMID- 15337091 TI - Plant response regulators implicated in signal transduction and circadian rhythm. AB - The so-called 'response regulators' were originally discovered as common components of the widespread histidine (His)-->aspartate (Asp) phosphorelay signal transduction system in prokaryotes. Through the course of evolution, higher plants have also come to employ such prokaryotic response regulators (RRs) for their own signal transduction, such as the elicitation of plant hormone (e.g. cytokinin) responses. Furthermore, plants have evolved their own atypical variants of response regulators, pseudo response regulators (PRRs), which are used to modulate sophisticated biological processes, including circadian rhythms and other light-signal responses. Recent studies using the model plant Arabidopsis thaliana have begun to shed light on the interesting functions of these plant response regulators. PMID- 15337092 TI - A global view of defense gene expression regulation--a highly interconnected signaling network. AB - Results from global mRNA expression profiling have revealed that plants express similar sets of defense mechanisms in response to different pathogens. Resistance is regulated by shifts in the balance among defense mechanisms, and by quantitative and/or kinetic enhancements that make the defense response more effective. The signaling mechanism that controls the activation of defenses consists of a highly interconnected network. Careful combination of recently developed technologies promises to deepen our understanding of the behavior of this complex signaling network. PMID- 15337093 TI - MicroRNA regulation of gene expression in plants. AB - It has only been a few years since we began to appreciate that microRNAs provide an unanticipated level of gene regulation in both plants and metazoans. The high level of complementarity between plant microRNAs and their target mRNAs has allowed rapid progress towards the elucidation of their varied biological functions. MicroRNAs have been shown to regulate diverse developmental processes, including organ separation, polarity, and identity, and to modulate their own biogenesis and function. Recently, they have also been implicated in some processes outside of plant development. PMID- 15337094 TI - Chromatin-based silencing mechanisms. AB - Eukaryotic genomes are organized into regions of transcriptionally active euchromatin and transcriptionally inactive heterochromatin. In plant genomes, heterochromatin is marked by methylation of cytosine and methylation of histone H3 at lysine 9. Heterochromatin formation is targeted to transposons as a means of defending the host genome against the deleterious effects of these sequences. Heterochromatin is directed to transposon sequences by transposon-derived aberrant RNA species and functions to prevent unwanted transcription and movement. Formation of heterochromatin at rRNA-encoding genes and centromere associated repeats might also involve an RNA-based mechanism that is designed to stabilize these potentially labile structures. PMID- 15337095 TI - ROP/RAC GTPase: an old new master regulator for plant signaling. AB - The ROP family of small GTPases has emerged as a versatile and pivotal regulator in plant signal transduction. Recent studies have implicated ROP signaling in diverse processes ranging from cytoskeletal organization to hormone and stress responses. Acting as a switch early in signaling cascades, ROPs are also capable of orchestrating several downstream pathways to amplify a specific signal. PMID- 15337096 TI - Guard cells: a dynamic signaling model. AB - The year 2003 has provided a continuing accretion of knowledge concerning the diverse ways in which guard cells sense and respond to abscisic acid. A deeper understanding of the biochemical mechanisms governing the response of guard cells to blue light has been gained, and new insights have been garnered regarding roles of the extracellular matrix in stomatal regulation. PMID- 15337097 TI - NPR1, all things considered. AB - Recent work has shown that the Arabidopsis NPR1 protein not only plays an essential role in salicylic acid (SA)-mediated systemic acquired resistance and rhizobacterium-triggered induced systemic resistance, but also is involved in crosstalk inhibition of jasmonic acid (JA)-mediated defense responses. Molecular characterization has revealed that activation of NPR1 and certain TGA transcription factors occurs under the reducing conditions that follow an initial oxidative burst after the induction of defense responses. In addition to NPR1 and TGA, the single-stranded DNA-binding transcription factor AtWhy1 and the WRKY70 transcription factor were recently found to be involved in SA-mediated defense and SA-JA crosstalk, respectively. PMID- 15337098 TI - Auxin signals--turning genes on and turning cells around. AB - The extremely wide spectrum of the plant processes that are influenced by auxin raises the question of how signals conveyed by a single molecule can trigger such a variety of responses. Although many aspects of auxin function remain elusive, others have become genetically tractable. The identification of crucial genes in auxin signal transduction and auxin transport in the past few years has led to molecularly testable concepts of how auxin signals regulate gene activities in individual cells, and how the polar transport of auxin could impact on patterning processes throughout the plant. PMID- 15337099 TI - Phytochrome-mediated light signalling in Arabidopsis. AB - The phytochrome photoreceptors regulate all major transitions during the life cycle of plants. The role of each member of the phytochrome family in Arabidopsis is starting to be understood, and a molecular description of phytochrome regulated flowering time and shade avoidance is emerging. Recent publications have challenged some areas of well-accepted models concerning phytochrome signalling. Moreover, the importance of proteolysis during phytochrome signalling is becoming very apparent. PMID- 15337100 TI - The autonomous pathway: epigenetic and post-transcriptional gene regulation in the control of Arabidopsis flowering time. AB - Mechanisms that mediate the control of flowering time have been accessed through a molecular genetic approach in Arabidopsis. Flowering is regulated by different pathways and, in the past year, all of the known components of the so-called autonomous pathway have been identified. The autonomous pathway comprises a combination of factors involved in RNA processing and epigenetic regulation that downregulate the floral repressor, FLOWERING LOCUS C (FLC). However, components of the autonomous pathway are more widely conserved in plant species other than Arabidopsis than is FLC. Therefore, the broadest lessons we learn from dissecting the function of the autonomous pathway may be in revealing how precision in regulated gene expression is delivered. PMID- 15337101 TI - PINning down the connections: transcription factors and hormones in leaf morphogenesis. AB - The past year has seen major advances in delineating the regulatory circuits that control leaf morphogenesis. Regulation of auxin transporters is pivotal in generating auxin gradients that define the sites of leaf inception. In addition, auxin acts via the ARGOS protein to regulate leaf size. HD-ZIPIII transcription factors controlled by micro-RNAs are required to specify adaxial identity in the leaf and vasculature. Elaboration of final leaf shape also depends on the regulated arrest of cell division, and the TCP and JAGGED transcription factors are key regulators of this process. Furthermore, the juxtaposition of abaxial and adaxial identities regulates leaflet placement in dissected leaf species. PMID- 15337102 TI - Network building: transcriptional circuits in the root. AB - Genes that control cell fate in all major root tissues have been described in Arabidopsis and new candidates that pattern the root apical meristem have been identified. Several phytohormones have long been known to affect root growth; it now appears that they have at least one common entry point into the transcriptional networks that regulate root growth. A map of gene expression in the Arabidopsis root has identified many new candidate genes that feed into and out of these transcriptional controls. PMID- 15337104 TI - Cytokinesis: lines of division taking shape. AB - Plant cytokinesis requires an orchestrated interplay of membrane and cytoskeleton dynamics, which results in the formation of the membrane that partitions the cytoplasm of the dividing cell. Until recently, phragmoplast-assisted cytokinesis of somatic cells was regarded as mechanistically different from 'non conventional' modes of cytokinesis, such as endosperm cellularisation or male meiotic cytokinesis. However, features that are similar among these diverse modes of cytokinesis have now been revealed by electron tomography, suggesting common underlying mechanisms that are also supported by genetic and molecular studies. Further insight into the complex process of cytokinesis has been gained from the identification of new components and from the analysis of known components. PMID- 15337103 TI - Signalling by tips. AB - New molecules, including protein kinases, lipids and molecules that have neurotransmitter activities in animals have emerged as important players in tip growing cells. Transcriptomics analysis reveals that the largest single class of genes expressed in pollen tubes encode signal transducers, reflecting the necessity to decode complex and diverse pathways that are associated with tip growth. Many of these pathways may use common intracellular second messengers, with ions and reactive oxygen species emerging as two major common denominators in many of the processes involved in tip growth. These second messengers might influence the actin cytoskeleton through known interactions with actin-binding proteins. In turn, changes in the dynamic properties of the cytoskeleton would define the basic polarity events needed to shape and modify tip-growing cells. PMID- 15337105 TI - Systems approaches to understanding cell signaling and gene regulation. AB - The age of 'omics' is upon us, and scientific papers that reflect this are starting to appear at an ever-increasing rate. The amount of information generated in any 'omics' program is daunting and often overwhelms plant scientists whose main interests relate to cell or developmental biology. For this revolution in data generation to have any impact in plant signaling studies, we must have great confidence in both the quality of the data and our ability to represent it in ways that are meaningful to general plant biologists. Systems biology has begun to address these issues and to provide examples in which the analysis of large data sets has led to biological insights into cell signaling and gene regulation. PMID- 15337106 TI - Nasal anatomy and physiology. AB - The anatomy and physiology of the internal nose is not always given appropriate consideration when rhinoplastic surgery is performed. Knowledge of the basic physiologic mechanisms of normal nasal function should be a prerequisite to operating on the external nasal structures because of the close relationship between form and function. Although the goals of rhinoplastic surgery are largely to enhance the cosmetic appearance of the visible portion of the nose, it is incumbent upon the surgeon to respect and preserve the critical roles of those portions that remain unseen. PMID- 15337107 TI - Acoustic rhinometry and its uses in rhinology and diagnosis of nasal obstruction. AB - Acoustic rhinometry is a tool that can aid in the assessment of nasal obstruction. The test is noninvasive, reliable, convenient, and easy to perform. Common clinical and practical uses of acoustic rhinometry for the rhinologic surgeon include assessment of"mixed" nasal blockage, documentation of nasal alar collapse, and preoperative planning for reduction rhinoplasty. Acoustic rhinometry can also be used to document the positive effect of surgery on nasal airway obstruction. PMID- 15337108 TI - Inflammatory sinonasal disease. AB - Inflammation of the nose and sinuses is a clinical problem frequently encountered in the practice of otolaryngology. The most common of these inflammatory illnesses are allergic rhinitis and acute and chronic rhinosinusitis. The recognition of inflammatory sinonasal diseases and an understanding of their pathophysiology are essential to successful management of these common illnesses. PMID- 15337109 TI - Rhinologic pharmacotherapy in rhinoplasty. AB - Rhinoplasty surgeons inevitably encounter therapeutic considerations in managing their patients, and a thorough understanding of nasal function, as well as of disorders of the nose and sinuses, is requisite for positive clinical outcomes. Patients suffering from allergic rhinitis are not precluded from undergoing rhinoplasty, whereas other disease processes may warrant an otolaryngologic evaluation before proceeding with surgery. A thorough medical history and examination elucidate sinonasal disease conditions that may not improve after septorhinoplasty but potentially could respond to pharmacologic therapy. PMID- 15337110 TI - Concurrent functional endoscopic sinus surgery and rhinoplasty: pros. AB - Despite historical concerns about the spread of infection from the sinuses to the nasal tissues,concurrent septorhinoplasty and endoscopic sinus surgery may be performed safely in most patients who meet the criteria for sinus surgery. However, otolaryngologists should use good medical judgment in selecting patients appropriate for the combined procedures.Patients with extensive sinus pathology or systemic illness are not the ideal candidates for concurrent surgery. It is recommended to perform the septal and sinus surgery first, so that the surgeon may postpone the elective rhinoplasty procedure if unfavorable intraoperative circumstances develop. Overall, when performed in carefully selected patients, contemporaneous rhinoplasty and endoscopic sinus surgery is safe and effective and offers many advantages for the patient. PMID- 15337111 TI - Considerations against concurrent functional endoscopic sinus surgery and rhinoplasty. AB - Over the last decade, functional endoscopic sinus surgery (FESS) has become the surgical modality of choice in the treatment of inflammatory sinus disease refractory to medical therapy. During the same interval, interest in cosmetic surgery procedures, including rhinoplasty, has exploded. Some of the published literature endorses the performance of concurrent FESS and rhinoplasty. This article addresses some of the concerns regarding the combined approach and its limitations. It offers the reader a framework for surgical risk assessment when concurrent procedures are considered. PMID- 15337112 TI - Considerations in the etiology, treatment, and repair of septal perforations. AB - Nasal septal perforations present a distinct challenge to the facial plastic surgeon. A clearcut causal factor must be established from a long, diverse list of potential causes. Surgical repair presents a complex technical challenge, because a septal perforation is a hole in three distinct contiguous layers composed of both right and left septal mucoperichondral flaps and the intervening cartilage, all three of which must be separated from each other and repaired individually. This article describes presenting symptoms and findings for septal perforations, the history and physical examination, causes, helpful hints for prevention of perforations, and surgical and nonsurgical treatment options and outcomes. PMID- 15337113 TI - The assessment and treatment of nasal obstruction after rhinoplasty. AB - Complaints of nasal obstruction after rhinoplasty and nasal surgery are disheartening for both the patient and the surgeon. When a patient presents with complaints of nasal obstruction after rhinoplasty and nasal surgery, the surgeon must reassess the history of symptoms and physical attributes contributing to nasal airway narrowing. The patient's expectations from surgery must also be discussed. The goal is total management of mucosal and anatomic contributors to nasal obstruction. This article describes various causes of nasal obstruction so as to provide rhinoplasty surgeons with the background needed to enhance their preoperative evaluations and avoid future surgical complications. PMID- 15337114 TI - Smell and taste disorders. AB - Disorders of taste and smell can present a challenge to the facial plastic surgeon. Obtaining a detailed history and examination is the key to the diagnosis and work-up of olfactory and gustatory dysfunction. Easy-to-administer tests are available for olfactory evaluation(University of Pennsylvania Smell Identification Test) and gustatory (taste sticks, tasting tablets) evaluation. The prognosis and management of olfactory and gustatory disease depend on its etiology. Despite ongoing research, the treatment of the disorders of smell and taste is limited. PMID- 15337116 TI - Mechanism of cell-cycle control: ligating the ligase. AB - The F-box protein SKP2 promotes the G1-S transition by targeting key regulators for proteasomal degradation via its capacity to function as the specificity factor for the SKP1 Cullin F-box SCF(SKP2) ubiquitin ligase. SKP2 is a labile protein, the levels of which oscillate in a cell cycle-dependent manner. SKP2 accumulation is often deregulated in cancer, which indicates that temporal control of SKP2 is essential for normal cell proliferation. Two new studies now suggest that SKP2 accumulation is determined by a second ubiquitin ligase, the anaphase-promoting complex or cyclosome, APC/C(CDH1). These studies highlight a novel mechanism wherein mitotic machinery communicates with proteins that regulate G1 phase progression. PMID- 15337117 TI - Coiled coils meet the chaperone world. AB - Coiled coils are versatile structural modules that engage in a variety of cellular activities. Recent studies illuminate their role as substrate-binding elements in the chaperone cofactor prefoldin and in the AAA+ ATPases involved in protein (un)folding processes. The use of coiled coils to mediate the binding of non-native proteins represents a novel strategy in chaperone design and a new function for coiled coils. PMID- 15337118 TI - NCD3G: a novel nine-cysteine domain in family 3 GPCRs. AB - The NCD3G [for nine-cysteine domain of family 3 G-protein-coupled receptors (GPCRs)] domain is a novel protein domain that is conserved in family 3 GPCRs, including metabotropic glutamate receptors, calcium-sensing receptors, pheromone receptors and taste receptors, with the exception of GABA(B) receptors. The NCD3G domain contains nine highly conserved cysteine residues. Structural predictions suggest that NCD3G might possess four beta strands and three disulfide bridges. The structural model of NCD3G highlights the conserved residues co-segregated with certain familial diseases. PMID- 15337119 TI - Common logic of transcription factor and microRNA action. AB - Over the past few years, microRNAs (miRNAs) have emerged as abundant regulators of gene expression. Like many transcription factors (TFs), miRNAs are important determinants of cellular fate specification. Here I provide a conceptual framework for miRNA action in the context of creating cellular diversity in a developing organism, and emphasize the conceptual similarity of TF- and miRNA mediated control of gene expression. Both TFs and miRNAs are trans-acting factors that exert their activity through composite cis-regulatory elements that are 'hard-wired' into DNA or RNA. TFs and miRNAs act in a largely combinatorial manner - that is, many different TFs or miRNAs control one gene - and they act cooperatively on their targets - that is, there are several cis-regulatory elements for a single TF or miRNA species in a target gene. Just as the set of TFs in a given cell type has been proposed to constitute a 'code' that specifies cellular differentiation, so 'miRNA codes' are likely to have conceptually similar roles in the specification of cell types. PMID- 15337120 TI - Ancestral lipid biosynthesis and early membrane evolution. AB - Archaea possess unique membrane phospholipids that generally comprise isoprenoid ethers built on sn-glycerol-1-phosphate (G1P). By contrast, bacterial and eukaryal membrane phospholipids are fatty acid esters linked to sn-glycerol-3 phosphate (G3P). The two key dehydrogenase enzymes that produce G1P and G3P, G1PDH and G3PDH, respectively, are not homologous. Various models propose that these enzymes originated during the speciation of the two prokaryotic domains, and the nature (and even the very existence) of lipid membranes in the last universal common ancestor (cenancestor) is subject to debate. G1PDH and G3PDH belong to two separate superfamilies that are universally distributed, suggesting that members of both superfamilies existed in the cenancestor. Furthermore, archaea possess homologues to known bacterial genes involved in fatty acid metabolism and synthesize fatty acid phospholipids. The cenancestor seems likely to have been endowed with membrane lipids whose synthesis was enzymatic but probably non-stereospecific. PMID- 15337121 TI - Division of labor in polycomb group repression. AB - The epigenetic maintenance of gene expression patterns is essential for developing and maintaining the diverse types of cell that cooperate to form the larger organism. Recent data suggest that proteins of the Polycomb group (PcG) use a combination of posttranslational modifications and structural changes to the underlying chromatin structure to maintain silenced epigenetic states. We are now beginning to understand the mechanisms by which the PcG proteins are able to silence genes and to maintain this silencing over many cell divisions. PMID- 15337122 TI - Caspases, IAPs and Smac/DIABLO: mechanisms from structural biology. AB - Caspases are the central component of the apoptotic machinery that irreversibly commits a cell to die. Whereas all caspases are structurally similar, those involved in apoptosis can be categorized functionally as either initiator or effector caspases, which are activated by distinct mechanisms. The activated caspases are subject to inhibition by the inhibitor of apoptosis family of proteins. This inhibition can be removed by Smac/DIABLO during apoptosis. The underlying molecular mechanisms of caspase regulation are discussed in this article. PMID- 15337123 TI - Phosphoryl group transfer: evolution of a catalytic scaffold. AB - It is proposed that enzymic phosphoryl-transfer reactions occur by concerted, step-wise, associative (phosphorane-intermediate) or dissociative (metaphosphate intermediate) mechanisms, as dictated by the catalytic scaffold and the reactants. During the evolution of a phosphotransferase family, the mechanism of the phosphoryl-transfer reaction is in constant flux, potentially changing with each adaptation of the catalytic scaffold to a new phosphoryl-donor-acceptor pair. Phosphotransferases of the recently discovered haloacid dehalogenase superfamily of enzymes, one of the largest and most ubiquitous of the phosphotransferase families characterized to date, are described in the context of the co-evolution of the catalytic scaffold and mechanism. PMID- 15337124 TI - P marks the spot: site-specific integrin phosphorylation regulates molecular interactions. AB - Integrins are heterodimeric adhesion receptors at the cell membrane that function as two-way signaling devices. The short intracellular tails of integrins are devoid of catalytic activity, but are nevertheless important for adhesion and signaling, presumably, through interactions with cytoplasmic molecules. Recently, the structure of the intracellular tails has been investigated using NMR, giving important new insight into how integrins might be regulated, but many questions remain unanswered. Signaling by many cell-surface receptors involves protein phosphorylation; over the past few years, phosphorylation of the integrin tails at specific sites has started to emerge as a dynamic mechanism that regulates molecular interactions between integrins and cytoplasmic molecules. This phosphorylation might give rise to signaling specificity and fine-tuning of the integrin-mediated responses. PMID- 15337125 TI - The pathogenesis of primary progressive multiple sclerosis: antibody-mediated attack and no repair? AB - Primary progressive multiple sclerosis (MS) differs from the more common form of MS which has an initial relapsing-remitting course in a number of ways, including pathological features, clinical course, differential diagnosis and response to treatment. The lesions in primary progressive MS tend to be more diffuse, less inflammatory and less likely to remyelinate than those occurring in relapsing remitting MS and secondary progressive MS; there are also fewer focal lesions in the brain in primary progressive MS. Recent evidence suggests that antibodies to central nervous system (CNS) antigens have an important role in disease progression. Such antibodies could cause demyelination, inhibit remyelination and cause axonal destruction. Ongoing immune attack by autoantibody and lack of CNS repair could be responsible for the gradually increasing disability in primary progressive MS. Further research on the B-cell and autoantibody response in primary progressive MS might lead to advances in diagnosis and treatment. Inhibition of autoantibody production by inducing B-cell apoptosis with rituximab is a potential new therapy for primary progressive MS. PMID- 15337126 TI - Anaesthesia for neurosurgery in the sitting position. AB - The sitting position offers many advantages in terms of surgical access for posterior fossa and posterior cervical spine surgery. However, these advantages must be balanced against the risks which include venous and paradoxical arterial air embolism, cerebral and myocardial ischaemia secondary to hypotension, and complications of the positioning itself. These are largely in the domain of the neuroanaesthetist. In this paper, therefore, we will review the advantages, disadvantages and management of complications of the sitting position, from the neuroanaesthetist's perspective. PMID- 15337127 TI - The antiepileptic Materia Medica of Pediacus Dioscorides. AB - Since it was written about the middle of the 1st Century AD, and up to comparatively recent times, the great Herbal, or Materia Medica, of Dioscorides provided medicine with its chief source of information about what were then considered therapeutic substances. The work contained data on various materials of botanical, biological and mineral origin which were claimed to provide benefit to sufferers from epilepsy, though often with no clear underlying rationale for their use. Some of these materials continued to be used as antiepileptic remedies over many centuries till they were finally recognised to be without useful effect in the disorder. The longest survivor amongst the Dioscoridean antiepileptic remedies was a rather esoteric one, viz. two stones taken from the belly of a young swallow during the rising phase of the moon and also whilst the swallow's parent birds were absent from the nest. The stones, or one of them, were worn against the skin of the seizure sufferer. The use of the swallow stones for epilepsy was recommended as late as in the writings of Thomas Willis (1675). PMID- 15337128 TI - Significance of chronic epilepsy in glial tumors and correlation with surgical strategies. AB - PURPOSE: This study was designed to compare the frequency of postoperative epilepsy in patients with acute and chronic pre-operative epilepsy and with frontal or temporal lobe glial tumors based on the hypothesis that patients with chronic epilepsy do worse. METHODS: We compared the clinical and diagnostic characteristics of the patients (n = 73) who had seizures preoperatively with those of the patients (n = 153) who did not. Among those who have had seizures preoperatively, we compared those (n = 32, chronic seizure group) who had seizures a year or more prior to surgery with those (n = 41, acute seizure group) who had seizures less than a year prior to surgery. RESULTS: Among the various factors, the frequency of benign pathology and favorable neurological state were higher in the seizure group than in the non-seizure group (p < 0.05). Complex partial seizures and low-grade tumors were frequent in the chronic seizure group, whereas simple partial seizures and high-grade tumors were frequent in the acute seizure group. Seizure-free rate was significantly higher in the acute seizure group than in the chronic group (p < 0.05). Also, the difference of seizure control rate between surgical strategies was statistically significant (p < 0.05). CONCLUSION: This study indicates that preoperative seizure duration and frequency have a close relationship with the frequency of postoperative epilepsy in patients with glial tumors. A longer duration may allow the formation of epileptogenic foci, leading to chronic epilepsy, and eventually have a negative effect on the prognosis of the patients. Factors including histopathological characteristics of the tumor, its location, seizure duration/frequency, and symptomatology should be taken into account when deciding on surgical strategies. PMID- 15337129 TI - Early post-operative seizures after burr-hole drainage for chronic subdural hematoma: correlation with brain CT findings. AB - The incidence of seizures in patients undergoing burr-hole crainiostomy with closed-system drainage for chronic subdural hematoma (CSDH) is low. The post operative use of anticonvulsants is, thus, controversial. In this study, we tried to correlate pre-operative computed tomographic (CT) appearance of the CSDH with the need for post-operative seizure prophylaxis. From April 1998 to November 2001, 128 cases of CSDH surgically treated at our hospital were studied. All patients underwent burr-hole craniostomy with closed system drainage. All CSDHs were classified as low-density, isodense, and mixed-density lesions according to CT findings. The incidence of early post-operative seizures (within 3 weeks of surgery) among all patients was 5.4% (7/128). In the subgroups by lesion density, the incidences were 6.2% (1/16) in the low-density group, 2.4% (2/83) in the isodense group, and 13.7% (4/29) in the mixed-density group (all p < 0.05). The mean age among the seven patients (five males and two females) who had seizures was 71 years. The locations of the CSDHs among the 128 patients were the left side of the brain in 53 (41.4%) patients, right side in 45 (35.2%), and bilateral in 30 (23.4%) patients. Among the seven patients who suffered from post-operative seizures, five (71.4%) had left side CSDHs, one (14.2%) had a right side CSDH, and one (14.2%) had bilateral CSDHs. We concluded that the post-operative seizure rate appeared high in the group with mixed-density type lesions on CT, and in those with left unilateral CSDH. We suggest the use of prophylactic anticonvulsants for patients with mixed-density lesions on pre-operative CT. PMID- 15337130 TI - Coagulopathy as a parameter to predict the outcome in head injury patients- analysis of 61 cases. AB - The correlation of coagulopathy and pupillary light reflex, the degree of midline shift in brain computer tomography and Glasgow outcome scale (GOS) after head injury were prospectively evaluated. From September 2002 to March 2003, 61 patients (45 males and 16 females; mean age: 41.9 years) after head injury were enrolled in the study. A modified coagulopathy score (CS) defined by prothrombin time, partial thromboplastin time, platelet count, D-dimer and fibrinogen was calculated for each patient within 24 h after injury. The CS was 2.3+/-2.7 (mean+/-SD). The incidence of abnormal coagulation following head injury in non survival cases was 100% and in survival cases 66%. The mortality rate was significantly increased to 75% in CS above 4 and 100% if CS was 6 or greater. The increase of D-dimer concentration appears to be common yet abnormal platelet counts are relatively uncommon among head trauma patients. Within 4 h after head injury, there is an initial hypercoagulable stage followed by hypocoagulable stage 6 h after head injury. Our results showed pupillary light reflex has the most significant correlation to GOS (rho = 0.727, p < 0.0001). It also reveals that coagulopathy score > or 4 (positive predictive value 90%) may have higher degree of accuracy to predict mortality comparing to both pupils being fixed or brain CT midline shift > or = 15 mm. We conclude that: (1) Coagulation state in head injury patients within 24 h after injury is of value in determining the outcome. (2) Coagulopathy score > or = 4 is a good predictor to evaluate mortality rate of head injury patients. PMID- 15337131 TI - Adjuvant gamma knife radiosurgery after meningioma resection. AB - We evaluated the usefulness of adjuvant treatment with gamma knife radiosurgery following meningioma surgery. During the past 8 years, we operated on 78 patients with meningiomas. Among these, 28 patients (36%) received gamma knife radiosurgery postoperatively. The indications for radiosurgery were as follows: residual tumour after surgery in 13 patients (46.4%), regrowth of residual tumour during the follow-up period in 7 patients (25%), recurrence after total removal (Simpson grade 1 or 2) in 7 patients (25%), and another intracranial meningioma in one patient (3.6%). The tumour diameter at the time of radiosurgery ranged from 5.2 to 48.1 mm (median 21.6 mm). Larger tumours with a mean diameter above 40 mm in two patients were treated with two-staged radiosurgery. The tumor marginal dose ranged from 8 to 23 Gy (median 12 Gy). The follow-up period was 3 84 months (median 30 months) after radiosurgery. The tumour size decreased in 17 patients (60.7%), remained unchanged in 9 patients (32.1%), and increased in 2 patients (7.2%). No radiation injury was experienced. Adjuvant therapy using gamma knife radiosurgery for meningiomas can achieve control of tumour growth and may improve patient outcomes. Careful surgical planning and follow-up are required to understand the usefulness and limitations of radiosurgery in this setting. PMID- 15337132 TI - Upper thoracic sympathectomy for axillary osmidrosis or bromidrosis. AB - The difference between axillary osmidrosis (AO) and axillary bromidrosis (AB) is the degree of odor and quantity of sweat, which is associated with selection of therapeutic modality theoretically. Upper thoracic sympathectomy has been used for both diseases but its effect needs to be further evaluated with more clinical data. We collected 108 patients with AO or AB treated by upper thoracic sympathectomy from July 1995 to July 2002. Of these patients, 42 suffered AO alone, 17 had AB (AO with axillary hyperhidrosis [AH]), and 49 had AO with palmar hyperhidrosis (PH). Ninety-two patients (183 sides) received anterior subaxillary transthoracic endoscopic sympathectomy (TES) and 17 patients (33 sides) received posterior percutaneous thoracic phenol sympathicolysis (PTPS). The levels of sympathectomy or sympathicolysis were T3-4 for AO and AB, and T2-4 for AO with PH. Mean follow-up period was 45.2 months (13-97 months). The satisfaction rates of patients were 52.4%, 70.6% and 61.2% for AO, AB and AO with PH, respectively. The rates of patients with improvement and satisfaction were 78.6%, 88.2% and 85.7% for AO, AB, and AO with PH, respectively. These results suggest that upper thoracic sympathectomy may be an acceptable treatment for AB or AO with PH rather than AO only. PMID- 15337133 TI - Muscular dystrophies and related skeletal muscle disorders in an Indian population--a prospective correlative study. AB - Since most centers in developing countries have limited facilities for investigation of patients with muscular dystrophies and similar disorders, this study was conducted with the aim of assessing the correlation between clinical, electromyographic (EMG) and histopathological findings in this group. We included 100 patients with muscular dystrophy and clinically similar disorders and subjected them to detailed clinical, electrophysiological and histopathological as well as immunohistochemical evaluation. Sensitivity, specificity, positive and negative predictive values and concordance rates for clinical and EMG diagnosis compared to diagnosis after histopathological examination were analyzed. With histopathology as standard, clinical diagnosis and a concordant EMG have very high sensitivity and negative predictive value (100%), but low specificity (33.3%). We conclude that detailed histopathological evaluation with immunohistochemical analysis is essential for the work-up of patients with suspected muscular dystrophies, since occasionally treatable muscle disorders like inflammatory myopathies can be detected when not suspected clinically. Muscle biopsies should only be conducted at major centers where full histochemical facilities are available. PMID- 15337134 TI - A pilot study of compassionate use of Levetiracetam in patients with generalised epilepsy. AB - Levetiracetam (LEV) has proven effective for partial seizures, suggesting the need to trial it in generalised epilepsy. Ten patients with generalised epilepsy were given compassionate use of LEV as a pilot study, attending 7 visits with seizure count (using diary) and compliance checked (pill count) with option for long term use. Seizure frequency was compared to baseline mean of the last 2 months and mean of follow-up. Patients were commenced on 500 mg I b.d, and titrated to a maximum of 3 g/day. There were 10 patients (7 females), aged 28-48, of whom 6 had primary generalised epilepsy (PGE) and 4 Lennox-Gastaut syndrome (LGS). At 7 month evaluation: 1 was seizure-free, 1 was 70% reduced, 3 were > or = 50% reduced, 2 were 30-35% reduced; 1 had no change; 1 was 10% increased and 1 was excluded because confounding pseudo seizures. Follow-up was 8-17 months (mean 13.8). The seizure-free patient became pregnant and had 2 seizures, but has been seizure-free for 2 months, at time of submission. A 16 months are three months seizure-free. One was 50% reduced at months 6 and 7, was 2 months seizure-free but then reverted to 50% per baseline. With respect to LGS, 1 withdrew due to aggression, 2 had 40% and 35% reduction at 13 and 15 months respectively and 1 had 25% increase (10% at 7 months). All patients were compliant. These data suggest that LEV may be effective for generalised epilepsy with a need for a larger clinical trial. PMID- 15337135 TI - Externalising deep brain electrodes: an increased risk of infection? AB - It is the practice in many centres to externalise deep brain electrodes in functional neurosurgery to confirm efficacy of therapy prior to full implantation of the pacemaker. It has been a concern that such practice might lead to an increased rate of infection. We report a retrospective study of the rates of infection in two major centres where all electrodes are externalised in one centre and directly implanted in the other. We have not found an increased rate of infection as a result of externalisation and feel, particularly in pain patients, that doing so can lead to significant cost savings by avoiding ineffective implantations. PMID- 15337136 TI - Screw implantation in spinous process for occipitoaxial fixation. AB - The feasibility of direct implantation of screws into the spinous process of the axis for fixation of plates in occipitoaxial stabilization is discussed. The method was used in 11 cases of congenital craniovertebral anomalies. The relatively strong and stubby spinous process of the axis in cases with occipitalization of the atlas facilitated screw implantation. In three cases two screws were implanted into the spinous process. The follow-up ranged from 3 to 43 months, the average being 26 months. During the period of follow-up one screw backed out from the spinous process. In the rest of the cases the screw in the spinous process satisfactorily fixed the axial end of the plate and stabilized the region. PMID- 15337137 TI - Operative technique: the anterior transcallosal transseptal interforniceal approach to the third ventricle and resection of hypothalamic hamartomas. AB - Background. We have previously described the resection of hypothalamic hamartomas (HH) using a transcallosal approach [Transcallosal resection of hypothalamic hamartomas, with control of seizures, in children with gelastic epilepsy, Neurosurgery, 2001]. Since then, we have refined the technique and now describe in detail an anterior transcallosal transseptal interforniceal approach to the third ventricle as a variation of the standard transcallosal interforniceal approach. The results of this series are presented to demonstrate the safety and efficacy of this approach. Method. HH were resected via an anterior transcallosal, transseptal, interforniceal approach to the third ventricle. This is a more anterior approach to the third ventricle with a more acute trajectory than has been described previously. Results. This approach provided excellent access to the floor of the third ventricle with minimal forniceal retraction and avoidance of dissection of the deep venous structures. Transcallosal resection of HH was performed in 45 patients aged 2.9-33 years (mean 11.3 years). Morbidity was minimal, including transient hemiparesis in 3, ongoing diabetes insipidus in 2, early short-term memory impairment in 16 (persistent in 6) and one patient developed pneumonia postoperatively but recovered. Conclusion. The anterior transcallosal transseptal interforniceal technique is an effective and relatively safe technique when used for the resection of HH. This operative approach is applicable to other pathology in the third ventricle or hypothalamic region and has advantages compared with the standard transcallosal approach to the third ventricle. PMID- 15337138 TI - Glioneuronal tumours in neurofibromatosis type 1: MRI-pathological study. AB - Neurofibromatosis type 1 (NF1) is an inherited disorder in which affected individuals develop both benign and malignant tumours at an increased frequency. Glioneuronal tumours, such as ganglioglioma and dysembryoplastic neuroepithelial tumour, have been previously reported in patients with NF1. We describe two patients with glioneuronal tumours and typical clinical features of NF1. Molecular analysis of these tumours did not demonstrate loss of the NF1 gene by fluorescence in situ hybridization (FISH) or immunohistochemistry analysis, suggesting they might not be causally associated with gross defects in NF1 expression. Because of the excellent prognosis following the resection of these tumours, it is important to distinguish them from other NF1-associated tumours. PMID- 15337139 TI - Images in neurosciences. PMID- 15337140 TI - Surface stimulation of the brain with a prototype array for a visual cortex prosthesis. AB - We are developing a neural prosthesis to electrically stimulate the visual cortex to restore basic visual perceptions to blind patients. The effects on cortical excitation of different stimulus configurations using a prototype electrode array are presented. Cats underwent a bilateral craniotomy to expose the cortex. An array for brain stimulation was placed on the surface of the right hemisphere. Cortical stimulation was undertaken in a variety of configurations while measuring the evoked responses that propagated through transcallosal pathways, at a homologous region on the contralateral hemisphere. Cortical excitation elicited by stimulation with a particular paradigm could be assessed by measuring the spatial spread and amplitudes of evoked responses in the contralateral hemisphere. Results from this transcallosal model have allowed us to examine the spatial and amplitude effects of cortical stimulation with our prototype electrode array and will aid in developing a neuroprosthesis for blind patients. PMID- 15337141 TI - Recurrent medulloblastoma--violation of Collin's law by 14 years. AB - We present a case of recurrent medulloblastoma, which violates Collin's law by 14 years. This is one of the longest exceptions to this rule published to date and serves as a reminder of the need for continued vigilance for many years after the original diagnosis. PMID- 15337142 TI - Glossopharyngeal neuralgia due to an epidermoid tumour in the cerebellopontine angle. AB - A 42-year-old female presented with typical glossopharyngeal neuralgia. Magnetic resonance imaging demonstrated an epidermoid tumour in the left cerebellopontine angle. Surgery showed that the tumour was compressing the glossopharyngeal nerve but no vessel was related to the nerve. The tumour was subtotally removed and after surgery the patient showed a complete relief of pain. This is a very rare case of glossopharyngeal neuralgia alone due to an epidermoid tumour. PMID- 15337143 TI - Refractory status epilepticus following self-poisoning with the organochlorine pesticide endosulfan. AB - We describe a case of refractory status epilepticus presenting to a rural general hospital in Sri Lanka. This patient's condition was precipitated by intentional self-poisoning with the organochlorine insecticide endosulfan. Although rarely seen in developed countries, pesticide poisoning particularly with endosulfan is an important cause of difficult-to-manage seizures in Asian countries. In this case report, we discuss the management of status epilepticus and refractory status epilepticus. Further, we specifically discuss the clinical pharmacology and toxicology of endosulfan. PMID- 15337144 TI - Superatentorial intracerebral hemorrhage following infratentorial surgery. AB - Supratentorial hematoma following infratentorial surgery is rare. We present two such patients with remote site supratentorial hematoma after posterior fossa surgery. In one patient, a supratentorial hematoma developed following surgery for an acoustic tumor. The supratentorial hematoma was located near where a supratentorial meningioma was excised five days before. No hematoma was seen on the immediate postoperative CT scan. In another patient there were two tumors, one in the pons and the other in the basal ganglia. This patient developed a basal ganglia hematoma following brain stem surgery. In both the patients, hematological profile revealed a coagulation abnormality following the posterior fossa surgery. Our first case stabilized conservative management, whereas the second required surgical evacuation of the hematoma. The differential diagnosis of declining level of consciousness after posterior fossa surgery must include supratentorial intracerebral hemorrhage and CT scan of the head is the diagnostic test of choice. PMID- 15337145 TI - Intracranial and dermatological cryptococcal infection in an immunocompetent man. AB - We report the case of a 72-year-old man with a history of Parkinson's Disease who presented with a history of falls, cognitive impairment and depressed mood. Neurological examination revealed moderate rigidity and bradykinesia. Alterations to his anti-Parkinsonian medication resulted in improvements in his mobility but no change in mental state. He was also noted to have developed multiple raised skin lesions. One of these lesions was biopsied and grew Cryptococcus neoformans. He subsequently had a cerebral MRI scan which showed enhancing lesions, in the left parietal lobe and right superior cerebellar peduncle, consistent with intracranial cryptococcal infection. Images of one of the skin lesions and the cerebral MRI scan are shown. PMID- 15337146 TI - Traumatic unilateral deep cerebral venous infarction. AB - A case of unilateral deep cerebral venous infarction in a young man associated with mild head injury is presented. The diagnosis is made on the basis of the clinical, radiological, operative and histopathological findings. We propose that a thrombosis or obstruction of the anterior segment of the basal vein of Rosenthal (BVR) was the primary pathology. This is discussed in respect to current knowledge of deep cerebral venous anatomy and embryology. PMID- 15337147 TI - Mycotic intracavernous carotid aneurysm. AB - Intracavernous carotid mycotic aneurysms are rare and management is dictated by clinical presentation. This case involved a patient presenting with a symptomatic expanding proximal internal carotid artery aneurysm treated with antibiotics and balloon occlusion but with thromboembolic complications resulting in a fatal outcome. Points of discussion include difficulties faced in reaching a diagnosis, management options for mycotic aneurysms and the rationale in this case for choosing endovascular rather than surgical treatment. The use and limitations of trial balloon occlusion are discussed as well as complications of vessel occlusion, in particular thromboembolism. Also discussed is the importance of surveillance imaging and the impact of sepsis on overall management. PMID- 15337148 TI - Unusual neuro-ophthalmic presentation of anterior communicating artery aneurysm with third nerve paresis. AB - Paresis of the oculomotor nerve associated with subarachnoid haemorrhage is considered a hallmark of aneurysms located at the junction of the internal carotid artery and posterior communicating artery. Third nerve palsy can also be caused by those aneurysms located in the intracavernous part of the internal carotid artery, basilar artery, posterior cerebral artery and superior cerebellar artery. However, oculomotor nerve paresis caused by an anterior communicating artery aneurysm is a very uncommon occurrence. We report a case of an elderly female with sudden severe headache who developed an acute third nerve paresis. Angiography revealed an anterior communicating artery aneurysm. Management and the pertinent literature are reviewed along with the mechanism of third nerve compression. PMID- 15337149 TI - Acute carotid arterial occlusion after burr hole surgery for chronic subdural haematoma in moyamoya disease. AB - Ischaemic strokes and neurological deterioration have been described after revascularisation surgery in patients with moyamoya disease, but accelerated acute occlusion of the internal carotid artery after burr hole surgery has not been reported in this setting. A 66-year-old woman with known moyamoya disease who presented with right motor weakness underwent burr hole surgery for a bilateral chronic subdural haematoma. Postoperatively, the patient had a crescendo transient ischaemic attack and then deteriorated. Angiography showed complete occlusion of the left internal carotid artery. Burr hole surgery may cause postoperative acute occlusion of a preexisting stenotic artery in patients with moyamoya disease. PMID- 15337150 TI - Dural metastases mimicking meningioma. Case report and review of the literature. AB - The typical appearance of meningioma on CT and MRI is well known. Particularly in the elderly, the imaging appearance is sometimes considered diagnostic of these benign tumours without histopathological confirmation. However, other more aggressive neoplasms can present with a classical CT and MRI appearance of meningioma, indicating the need for histopathological confirmation wherever possible. We report a case of dural metastases which, on both pre-operative CT and MRI and at surgery, had the typical appearance of a falcine meningioma. Histopathology and immunohistochemistry revealed adenocarcinoma of renal cell origin, and the renal primary was identified on subsequent abdominal investigation. The literature regarding dural metastases is reviewed. To our knowledge, this is the first reported case of a renal carcinoma metastasizing directly to the dura. Although rare, dural metastases can mimic meningioma, and this needs to be considered if conservative therapy or radiosurgery are to be offered to a patient with radiological diagnosis of meningioma. PMID- 15337151 TI - Intermittent claudication -- atypical presentation, diagnosis and treatment. AB - A unique case of a man with Fabry's disease and associated small vessel vasculopathy manifesting as recurrent episodes of intermittent claudication is described. The case highlights the concept that microvascular involvement due to local accumulation of glycosphingolipid in the smooth muscle fibres of vessel walls may be responsible for claudicant symptoms in such patients rather than the more classical macrovascular insufficiency. PMID- 15337152 TI - Normal pressure hydrocephalus after gamma knife radiosurgery for cerebellopontine angle meningioma. AB - There are no reports of hydrocephalus following radiosurgery for a meningioma. We report on a case where gamma knife therapy for a 4 cm diameter right cerebellopontine meningioma accelerated hydrocephalus three months post treatment. Examination of the cerebrospinal fluid (CSF) revealed a high protein level and thus, CSF malabsorption and CSF obstruction might have occurred after the radio surgery. It is important to consider this pathology, and the need for long term follow up. PMID- 15337153 TI - Intraventricular meningioma with drop metastases and subgaleal metastatic nodule. AB - We report a case of malignant intraventricular meningioma with CSF drop metastases and an implantation metastatic subgaleal nodule in a 53-year-old woman. Malignant intraventricular meningiomas are rare with only seven cases being reported in the literature. These tumours can be very aggressive and one should consider immediate postoperative radiotherapy. PMID- 15337154 TI - Non-traumatic spontaneous acute epidural haematoma -- report of two cases and review of the literature. AB - Epidural haematomas are usually associated with preceding head trauma. The entity of non-traumatic spontaneous acute epidural haematoma is rare and most commonly occurs in the presence of infectious disease. It can also occur in the presence of coagulopathy, vascular malformations of the dura mater and haemorrhagic tumours. Sickle cell disease, systemic lupus erythematosus, open heart surgery and haemodialysis have also been implicated as causative factors. The authors report two cases of spontaneous epidural haematomas (one of unknown aetiology and one from a coagulation disorder) and discuss the aetiological agents involved in this rarely described condition. PMID- 15337155 TI - Spontaneous intracerebral haemorrhage following evacuation of chronic subdural hematomas. AB - Spontaneous intracerebral haematoma (ICH) is an extremely unusual complication following the evacuation of a chronic subdural haematoma (CSDH). Good outcome is expected after the drainage of the CSDH and neurological deterioration is a cause for serious concern. Authors report three cases of spontaneous ICH away from the site of surgery following evacuation of a CSDH with a review of literature. Changes in cortical blood flow following decompression of a long standing CSDH may be responsible for the ICH. PMID- 15337156 TI - Dermoid of the conus medullaris. AB - Dermoids are rare midline tumours. When involving the spinal cord they usually present early, in the first decade of life. We present a case of an elderly male with paraparesis. His MRI showed cord expansion at the tip of the conus medullaris; contrast enhancement with Gadolinium revealed a cystic lesion and histopathology confirmed the diagnosis of a dermoid cyst. PMID- 15337157 TI - Letter to the editor. PMID- 15337158 TI - Multiple origins of replication in archaea. AB - Until recently, the only archaeon for which a bona fide origin of replication was reported was Pyrococcus abyssi, where a single origin was identified. Although several in silico analyses have suggested that some archaeal species might contain more than one origin, this has only been demonstrated recently. Two studies have shown that multiple origins of replication function in two archaeal species. One study identified two origins of replication in the archaeon Sulfolobus solfataricus, whereas a second study used a different technique to show that both S. solfataricus and Sulfolobus acidocaldarius have three functional origins. These are the first reports of archaea having multiple origins. This finding has implications for research on the mechanisms of DNA replication and evolution. PMID- 15337159 TI - Antibiotic-induced lateral transfer of antibiotic resistance. AB - As do many temperate bacteriophages, integrating conjugative elements (ICEs) recruit the SOS DNA damage response to mobilize themselves from the bacterial chromosome and infect other cells. This transfers resistance to multiple antibiotics. Several commonly used antibiotics induce the SOS response, potentially hastening genetic change and the evolution to resistance of pathogenic populations. The use of such antibiotics should be reconsidered. PMID- 15337160 TI - Exploring the boundaries of life. PMID- 15337161 TI - Phylogenetic reconstruction and lateral gene transfer. AB - Lateral gene transfer (LGT) is often seen as a form of noise, obscuring the phylogenetic signal with which we might hope to reconstruct the evolution of a group of organisms, or indeed the history of all life (the Tree of Life). Such reconstruction might still be possible if the subset of genes conserved among all genomes in a group (or common to all genomes) comprise a core that is relatively refractory to LGT. Several papers designed to test this notion have recently appeared, and here we re-analyze one, which claims that the core of single-copy orthologs shared by all sequenced genomes of the gammaproteobacteria is essentially free of LGT. This conclusion is unfortunately premature, and it is very hard to determine what fraction of this core has been affected by LGT. We discuss other difficulties with the core concept and suggest that, although the core idea must remain part of our understanding of phylogenetic relationships, it should not be the sole basis for defining such relationships, because these are not exclusively tree-like. We suggest instead a more complex but more natural framework for classification, which we call the Synthesis of Life. PMID- 15337162 TI - Human intestinal bacteria as reservoirs for antibiotic resistance genes. AB - Human intestinal bacteria have many roles in human health, most of which are beneficial or neutral for the host. In this review, we explore a more sinister side of intestinal bacteria; their role as traffickers in antibiotic resistance genes. Evidence is accumulating to support the hypothesis that intestinal bacteria not only exchange resistance genes among themselves but might also interact with bacteria that are passing through the colon, causing these bacteria to acquire and transmit antibiotic resistance genes. PMID- 15337163 TI - Host nutritional status: the neglected virulence factor. AB - The emergence of new infectious diseases and old diseases with new pathogenic properties is a burgeoning worldwide problem. Severe acute respiratory syndrome (SARS) and acquired immune deficiency syndrome (AIDS) are just two of the most widely reported recent emerging infectious diseases. What are the factors that contribute to the rapid evolution of viral species? Various hypotheses have been proposed, all involving opportunities for virus spread (for example, agricultural practices, climate changes, rainforest clearing or air travel). However, the nutritional status of the host, until recently, has not been considered a contributing factor to the emergence of infectious disease. In this review, we show that host nutritional status can influence not only the host response to the pathogen, but can also influence the genetic make-up of the viral genome. This latter finding markedly changes our concept of host-pathogen interactions and creates a new paradigm for the study of such phenomena. PMID- 15337164 TI - Intracellular bacterial communities of uropathogenic Escherichia coli in urinary tract pathogenesis. AB - Urinary tract infections in young, healthy women frequently recur, despite their traditional classification as acute infections. Conventional wisdom dictates that uropathogens causing recurrent infections in such individuals come from the fecal or vaginal flora, in the same manner as the initial infection. However, recent studies of uropathogenic Escherichia coli have found that it can carry out a complex developmental program within the superficial epithelial cells of the mouse bladder, forming intracellular bacterial communities with many biofilm-like properties. These intracellular biofilms allow the bacteria to outlast a strong host immune response to establish a dormant reservoir of pathogens inside the bladder cells. Re-emergence of bacteria from this reservoir might be the source of recurrent infection. PMID- 15337165 TI - hhLIM protein is involved in cardiac hypertrophy. AB - Proteins of the LIM family are critical regulators of development and differentiation in various cell types. Here we examined the roles of one new member of LIM family, hhLIM, in cardiac hypertrophic growth and cardiac muscle specific gene expression. To model the increase in endogenous hhLIM transcriptional activity that occurs in response to hypertrophic stimulation, hhLIM was overexpressed using a recombinant plasmid for hhLIM. The results showed that overexpression of hhLIM resulted in increased cell volume in both C2C12 muscle cells (>1.5-fold) and cardiac myocytes (>2.49-fold), a phenotype commonly associated with cardiac hypertrophy. RT-PCR and Western blot showed that transfection of hhLIM into C2C12 muscle cells and cardiomyocytes increased skeletal alpha-actin levels and triggered the expression of the embryonic-related gene BNP, which is associated with cardiac hypertrophy. Inhibition of hhLIM expression by antisense transcripts blocked the induction of skeletal alpha-actin and BNP expression by endothelin-1. These data indicated that hhLIM played a role in regulation of cardiomyocyte growth and cell size in response to hypertrophic stimuli through its modulation of skeletal alpha-actin and BNP expression. We also determined by confocal laser scanning microscopy and immunoprecipitation that hhLIM was associated with alpha-actin and localized in the cytoplasm in unstimulated cells, and was relocalized from the cytoplasm to the nucleus upon hypertrophic stimulation. These studies suggest that hhLIM protein is involved in cardiac hypertrophy. PMID- 15337166 TI - Dysregulation of apoptosis by benzene metabolites and their relationships with carcinogenesis. AB - Benzene is a widely recognized human carcinogen, the effect of which is attributed to the production of reactive oxygen species (ROS) from its metabolites. Although there have been many reports on the relationship between DNA damage induced by benzene metabolites and carcinogenesis, only a report approached the subject by examining the benzene-induced dysregulation of apoptosis. Inhibition of apoptosis, aberrantly prolonging cell survival, may contribute to cancer by facilitating the insurgence of mutations and by creating a permissive environment for genetic instability. In this study, we examined the mechanism of antiapoptotic effects by benzene metabolites, p-benzoquinone (BQ) and hydroquinone (HQ), and their relationships with carcinogenesis. BQ and HQ inhibited the apoptotic death of NIH3T3 cells induced by both serum starvation and lack of an extracellular matrix (ECM). An antioxidant agent, N acetylcysteine, significantly inhibited the antiapoptotic effects induced by benzene metabolites, indicating that the effects were mainly due to the production of ROS. Furthermore, BQ and HQ inhibited the in vitro caspase-3 activation, suggesting that the inhibition of caspase-3 activation due to ROS produced by BQ- and HQ-treatment was related to the suppression of apoptosis. The cells that escaped apoptosis could survive with the addition of serum and attachment to the ECM. Levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine were higher in the cells which survived after BQ- and HQ-treatment than in the normal cells. Furthermore, the cells treated with BQ and HQ showed greater proliferation than normal cells under low-serum conditions and anchorage-independent growth in soft agar. These findings suggested that benzene metabolites induced dysregulation of apoptosis due to caspase-3 inhibition, which contributes to carcinogenesis. PMID- 15337167 TI - Thermal unfolding of medium-chain acyl-CoA dehydrogenase and iso(3)valeryl-CoA dehydrogenase: study of the effect of genetic defects on enzyme stability. AB - Genetic defects affecting acyl-CoA dehydrogenases (ACAD)-key enzymes in the degradation of fatty acids and branched chain amino acids-are increasingly recognized as being more widespread than originally thought. For the medium-chain acyl-CoA dehydrogenase (MCAD), the K304E mutation is the most common genetic defect among Caucasian populations. The effect of substrate or substrate analog binding on the stability of wild-type MCAD and isovaleryl-CoA dehydrogenase (i3VD) and their genetic mutants (K304E- and T168A-MCAD and A282V-i3VD) is examined. Binding to the mutant ACADs is generally approximately 10-fold weaker compared to wild-type proteins. Thermal stability of wt-MCAD (melting point approximately 53.6 degrees C) is significantly higher compared to wt-i3VD ( approximately 49.3 degrees C). With the exception of the A282V-i3VD mutant, a high degree of stabilization (5-11 degrees C) is induced by conversion into the reduced enzyme form complexed with product. The results are discussed based on the 3D-structures of the enzymes, and it is concluded that in the case of K304E MCAD thermal stability as such is not a major contribution to the clinical phenotype. With the T168A-MCAD and A282V-i3VD mutants, however, the diminished thermal stability and minor stabilization by ligands must be regarded as an important factor contributing to the manifestation of the disease. PMID- 15337168 TI - Ultrastructural organization of ex vivo amyloid fibrils formed by the apolipoprotein A-I Leu174Ser variant: an atomic force microscopy study. AB - Atomic force microscopy was employed to study ex vivo amyloid material isolated from the transplanted hearts of two patients affected by systemic amyloidosis caused by the Leu174Ser apolipoprotein A-I variant. The purified material consists of fibrils and globular aggregates. For both patients the same morphological patterns are observed; in addition, fibril diameters obtained for the two patients turn out to be compatible, both in air (2.00+/-0.02 and 2.04+/ 0.04 nm) and under liquid (10.7+/-0.4 and 11.3+/-0.5 nm). Fibrils display heterogeneous morphologies, occasionally showing a left-handed twist. Inspection of fibril ends, the study of fibril contour shape and the analysis of partially unfolded fibrils yield independent evidences suggesting that most twisted fibrils are composed of three protofilaments. The size of globular aggregates is the same for both patients (4.4+/-0.4 and 5.1+/-0.5 nm, measured under liquid) and is compatible with the protofilament expected diameter, suggesting that globules may represent protofilament precursors. PMID- 15337169 TI - Left ventricular mitogen activated protein kinase signaling following polymicrobial sepsis during streptozotocin-induced hyperglycemia. AB - We hypothesized that sepsis during hyperglycemia would activate left ventricular (LV) mitogen activated protein kinase (MAPK) signaling mechanisms and modulate generation of endothelin-1 (ET-1) and nitric oxide (NO) that can contribute to the progression of LV dysfunction. A single injection of streptozotocin (STZ, 60 mg/kg, via tail vein) was used to produce type 2 diabetes in male SD rats. Polymicrobial sepsis and sham-sepsis were induced using single i.p. injection of cecal inoculum and sterile 5% dextrose water, respectively, on the 13th and 27th day following STZ injection. Both 2-week (2-wk) and 4-wk diabetes groups were associated with hyperglycemia and weight loss. LV end diastolic pressure (LVEDP) was significantly increased in 4-wk diabetes but not in 2-wk diabetes group. Plasma concentration of tumor necrosis factor-alpha (TNF-alpha) was significantly increased in 4-wk diabetes+sepsis group as compared to sham, 2-wk diabetes+sepsis and sepsis groups. Elevated plasma and LV ET-1 and NO byproducts (NOx) along with LV preproET-1 and inducible nitric oxide synthase (iNOS) protein expression were observed in 4-wk but not in 2-wk diabetes group. Sepsis further elevated LV iNOS and preproET-1 in 4-wk diabetes group. Up-regulated phosphorylation of LV p38 MAPK, extracellular signal-regulated kinase 1/2 (ERK1/2) and heat shock protein 27 (Hsp27) was observed in 4-wk diabetes group. Sepsis caused a factorial increase in LV p38-MAPK and Hsp27 phosphorylation and iNOS up-regulation but not ERK1/2 following progression from 2-wk to 4-wk diabetes. The study provides evidence that sepsis up-regulated LV iNOS, p38-MAPK phosphorylation and elevated LVEDP during 4-wk diabetes. We concluded that sepsis contributes in the development of LVEDP dysfunction and alteration in signaling mechanisms depending upon the progression from 2-wk to 4-wk diabetes in the rat. PMID- 15337170 TI - Extracellular matrix changes in early osteochondrotic defects in foals: a key role for collagen? AB - Osteochondrosis (OC) is the most important developmental orthopaedic disease in the horse. Despite some decades of research, much of the pathogenesis of the disorder remains obscure. Increasing knowledge of articular cartilage development in juvenile animals led to the presumption that the role of collagen in OC might be more important than previously thought. To study collagen characteristics of both cartilage and subchondral bone in young (5 and 11 months of age) horses, samples were taken of subchondral bone and articular cartilage from a group of 43 Dutch Warmblood foals and yearlings that suffered from varying degrees of OC. Based on a histological classification, lesions were graded as early, middle and end stage. Collagen content and some posttranslational modifications (lysyl hydroxylation, hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) cross links) were determined, as was proteoglycan content. Data were compensated for site effects and analysed for differences due to the stage of the lesion. In early lesions total collagen was significantly decreased in both cartilage and subchondral bone of 5- and 11-month-old foals. Also in cartilage, HP cross linking was reduced in the early lesions of 5- and 11-month-old foals, while LP cross-linking was decreased in subchondral bone of the end-stage lesions of both 5- and 11-month-old foals. Hydroxylysine content was unaffected. Collagen content remained reduced in cartilage from middle- and end-stage lesions, but returned to normal in subchondral bone. In cartilage there was a decrease in proteoglycan content in the end-stage lesions of both age groups. Thus, alterations of the collagen component, but not of the proteoglycan component, of the extracellular matrix might play a role in early OC. More severe lesions show a more general picture of an unspecific repair reaction. Biomarkers of collagen metabolism can be expected to be good candidates for early detection of OC. PMID- 15337171 TI - Transport of N-acetylaspartate via murine sodium/dicarboxylate cotransporter NaDC3 and expression of this transporter and aspartoacylase II in ocular tissues in mouse. AB - Canavan disease is a genetic disorder associated with optic neuropathy and the metabolism of N-acetylaspartate is defective in this disorder due to mutations in the gene coding for the enzyme aspartoacylase II. Here we show that the plasma membrane transporter NaDC3, a Na+-coupled transporter for dicarboxylates, is able to transport N-acetylaspartate, suggesting that the transporter may function in concert with aspartoacylase II in the metabolism of N-acetylaspartate. Since Canavan disease is associated with ocular complications, we investigated the expression pattern of NaDC3 and aspartoacylase II in ocular tissues in mouse by in situ hybridization. These studies show that NaDC3 mRNA is expressed in the optic nerve, most layers of the retina, retinal pigment epithelium, ciliary body, iris, and lens. Aspartoacylase II mRNA is coexpressed in most of these cell types. We conclude that transport of N-acetylaspartate into ocular tissues via NaDC3 and its subsequent hydrolysis by aspartoacylase II play an essential role in the maintenance of visual function. PMID- 15337172 TI - Role of nitric oxide increase on induced programmed cell death during early stages of rat liver regeneration. AB - We analysed the possible cellular mechanism involved in the NO action in the balance between apoptosis and cell proliferation in liver regeneration process. We determined p53, proapoptotic protein Bax, antiapoptotic Bcl-xL, proliferating cell nuclear antigen (PCNA) and apoptotic index at the early stages of regenerative process after NO increase by lipopolysaccharide-induction (LPS) of inducible-type nitric oxide synthase (iNOS) and by direct NO donor (sodium nitroprusside, SNP). Male Wistar rats were randomised in four experimental groups: sham operated control (Sh), partial hepatectomised control (PH-C), partial hepatectomised pretreated with LPS (2 mg/kg body weight, i.p.) (PH-LPS), and partial hepatectomised pretreated with SNP (2.5 mg/kg body weight, i.v. at a rate of 1 ml/h) (PH-SNP). Animals were killed 5 h post-surgery. Hepatic cytosolic iNOS showed an increase of 34% in PH-C animals with respect to Sh, and LPS treatment increased iNOS protein levels 30% compared with PH-C. Bax and p53 protein levels showed significant increases in LPS- and SNP-treated hepatectomised rats with respect to PH-C. The apoptotic indexes were increased 75% in both, PH-LPS and PH-SNP rats versus PH-C. The increase of NO did not show any change in the proliferation process. These results suggest that NO is involved in apoptosis via p53 and Bax proteins after PH, showing a tightly regulated growth process in liver regeneration. PMID- 15337173 TI - Electrochemical investigation into the redox activity of Fe(II)/Fe(III) in the presence of nicotine and possible relations to neurodegenerative diseases. AB - The biological relevance of Fe(II)/Fe(III) is becoming evermore apparent, especially in relation to its potential role in the progression of neurodegenerative diseases such as Parkinson's and Alzheimer's disease. The reported relationship between smoking and a reduced incidence of neurodegenerative disorders prompted this work. In order to investigate whether nicotine can interact with iron, we have studied the electrochemical behaviour of a Fe(II)/Fe(III) redox couple in the presence of nicotine. Solubility issues and lack of available nonreacting salts of nicotine necessitated studies being conducted at low pH values. Cyclic voltammetry experiments revealed a definite alteration in the electrochemical behaviour of the Fe(II)/Fe(III) redox couple suggesting the capability of nicotine to complex with free iron and, hence, reduce its reactivity. This is evident from a slower rate of heterogeneous electron transfer, ks, and a shift from reversible to quasi-reversible behaviour, as characterised from the diffusion coefficient (D), the full width half maximum (FWHM), DeltaEp and Ef. Additional complexation titrations, pH ranging from 1 to 7, confirm a weak complexation reaction occurring between Fe(III) and nicotine. PMID- 15337174 TI - The reduced insulin-mediated glucose oxidation in skeletal muscle from type 2 diabetic subjects may be of genetic origin--evidence from cultured myotubes. AB - Several defects in response to insulin have been described in vivo and in vitro in type 2 diabetes: a decreased glucose transport, defective glucose oxidation and altered glycogen synthesis. At present, it is unknown whether glucose oxidation is primarily affected or secondarily affected by, e.g. increased free fatty acids (FFA). The aim of this study was to evaluate whether myotubes established from type 2 diabetic subjects express a primarily or a FFA-induced reduced insulin-mediated glucose oxidation. We have therefore investigated glucose oxidation under basal, physiological conditions and during acute insulin stimulation with/without FFA. We found that myotubes established from type 2 diabetic subjects express a reduced insulin-stimulated increase in glucose oxidation. Moreover, an acute exposure to FFA reduces insulin-mediated glucose oxidation without alterations in glucose uptake and glycogen synthesis. Thus, we conclude that the diminished increase in insulin-stimulated glucose oxidation seen in type 2 diabetic subjects in vivo may be of genetic origin. Moreover, the glucose-fatty acid cycle seems not to be crucial for the pathophysiology of insulin resistance. PMID- 15337175 TI - Biological and biophysical principles in extracorporal bone tissue engineering. Part III. AB - Over the last decade extracorporal bone tissue engineering has moved from laboratory to clinical application. The restoration of maxillofacial bones from cell harvesting through product manufacture and end-use has benefited from innovations in the fields of biomechanical engineering, product marketing and transplant research. Cell/scaffold bone substitutes face a variety of unique clinical challenges which must be addressed. This overview summarises the recent state of the art and future anticipations in the transplantation of extracorporally fabricated bone tissues. PMID- 15337176 TI - Automated laser registration in image-guided surgery: evaluation of the correlation between laser scan resolution and navigation accuracy. AB - Markerless patient registration based on the facial skin surface makes logistics prior to image-guided surgery much easier, as it is not necessary to place and measure registration markers. A laser scan registration of the surgical site takes the place of conventional marker-based registration. In a clinical study, the stability and accuracy of markerless patient registration was evaluated in 12 patients. Intraoral titanium markers served as targets for the infrared-pointer of the navigation system in order to check the accuracy of the markerless registration process. The correlation between laser scan resolution and navigation accuracy was checked using seven different laser scan resolutions (a cloud of 300,000 laser scan points down to 3750 laser scan points of the surgical site). The markerless patient registration was successful as long as high laser scan resolution was used (30,000 laser scan points and more): the titanium markers were detected with a mean deviation of 1.1 +/- 0.2 mm. Low resolution laser scans (6000 laser scan points of the surgical site and less) revealed inaccuracies up to 6 mm. PMID- 15337177 TI - Stabilisation of sagittal split advancement osteotomies with miniplates: a prospective, multicentre study with two-year follow-up. Part III--condylar remodelling and resorption. AB - This prospective study implied a two-year follow-up in a group of patients that underwent a Bilateral Sagittal Split Osteotomy (BSSO) for advancement of the mandible that were treated in seven institutions following the same treatment protocol (using two miniplates). The aim of Part III of this study was to define a Condylar Morphology Scale (CMS) and to analyse radiological changes in the TMJ after BSSO in relation to postoperative relapse and to determine the incidence of morphologic changes and its risk factors. It was concluded that the used 3-point CMS served its purpose well. In eight patients (4%) resorption of the condyle developed postoperatively. The value of preoperative cephalograms to predict condylar alterations appeared to be limited (12% explained variance). Patients treated at a relative low age (< or = 14 years) appeared to be at risk for the occurrence of condylar alterations including resorption. A steep mandibular plane angle and the low facial height ratio (post:ant) were also significantly related to the occurrence of condylar alteration, but the multi variance regression showed that these parameters had only limited value. The occurrence of pain and TMJ sounds in the first few months postoperatively are highly suspicious for condylar changes to occur in the next months. PMID- 15337178 TI - Skeletal stability following maxillary impaction and mandibular advancement. AB - The purpose of this study was to retrospectively evaluate the stability of combined Le Fort I maxillary impaction and mandibular advancement performed for the correction of skeletal Class II malocclusion. Twenty-nine patients, mean age 22.6 years, underwent bimaxillary surgery with rigid internal fixation. Standardised cephalometric analyses were performed using serial lateral cephalometric radiographs. The post-surgical follow-up was a minimum of 12 months, with a mean of 25.2 months. The maxilla was impacted by a mean of 4.3 +/- 3.3 mm, and horizontally advanced by a mean of 2.6 +/- 2.3 mm. The results demonstrated that the maxilla tended to move anteriorly and inferiorly but this was not significant in either horizontal or vertical planes (P > 0.05). The mean advancement of the mandible, at menton, was 10.7 +/- 5.6 mm, and in 14 cases (48.2%) menton was advanced greater than 10 mm. In 34.7% of the patients the mandible underwent posterior movement between 2 and 4 mm. In the vertical plane, gonion moved superiorly by a mean of 2.7 +/- 3.6 mm which was significant. Significant mandibular relapse was found to have occurred in five female patients, with high mandibular plane angles who had undergone large advancements of greater than 10 mm. In conclusion, the majority of patients undergoing bimaxillary surgery for the correction of skeletal Class II malocclusions maintained a stable result. However, a small number of patients, exhibiting similar characteristics, suffered significant skeletal relapse in the mandible secondary to condylar remodelling and/or resorption. PMID- 15337179 TI - A retrospective evaluation of rigid fixation in orthognathic surgery using a biodegradable self-reinforced (70L:30DL) polylactide. AB - OBJECTIVE: This study was designed to evaluate clinical and radiological evidence of osteotomy site healing in orthognathic surgery after rigid fixation using a biodegradable plating system. STUDY DESIGN: A follow up of 30 patients who underwent orthognathic surgery using a biodegradable self-reinforced (70L:30DL) polylactide plating system was presented. The follow-up schedule for all patients consisted of regular appointments at 1-180 days after surgery. Clinical evaluation involved notation of any abnormal swelling, infection, discoloration, or discharge at the osteotomy sites. Stability was evaluated by manual palpation. For radiographic evaluation, panoramic radiographs were taken immediately after surgery, and again at 7-180 days. The radiographs were analyzed for any visual changes in osteotomy fragments, resorptive changes in osteotomy fragments, callus formation, and union of the osteotomy segments. RESULTS: No clinical complications and no radiological changes in the osteotomy sites were observed. Regarding the clinical usefulness of the biodegradable fixation system, fixation at the time of operation was considered as excellent in all 30 cases. CONCLUSIONS: The conclusions of this study were that self-reinforced (70L:30DL) polylactide was considered to be comparable to other forms of rigid internal fixation for orthognathic surgery. PMID- 15337180 TI - Intraoperative real-time genetic diagnosis for sentinel node navigation surgery. AB - Sentinel node navigation surgery (SNNS) has received considerable attention for its role in deciding whether to perform neck dissection in patients with early oral cancer. However, diagnostic accuracy and its intraoperative availability of results remain important concerns. First, we shortened the examination time required for genetic diagnosis. Second, we assessed the quality of the extracted mRNA. Third, 10 patients with early N0 oral cancer underwent SNNS, using our new technique for genetic diagnosis to determine whether neck dissection was required. The examination time of our one-step reverse-transcriptase polymerase chain reaction method using a minicolumn and LightCycler was successfully shortened to 2 h, permitting intraoperative genetic diagnosis. The extracted mRNA was of high quality. Six sentinel nodes in four patients were diagnosed to be metastatic on genetic diagnosis; these patients underwent neck dissection. The other six patients avoided unnecessary surgery. We conclude that intraoperative genetic diagnosis of micrometastasis holds promise of being a sensitive method that can be used to support SNNS. PMID- 15337181 TI - Changes in facial form relative to progressive atrophy of the edentulous jaws. AB - The surgical and prosthodontic rehabilitation of the edentulous patient aims to restore oral function and facial form. Planning treatment requires an understanding of the effect of progressive jaw atrophy, and the concomitant effect on the soft tissues of the face. This study examined 179 Caucasians at different stages of jaw atrophy according to the Cawood and Howell classification; various standard anthropological measurements of the face, according to Farkas, were also taken. We have demonstrated that changes in the soft tissues are related to the degree of underlying jaw atrophy. This has important implications when planning surgical and prosthodontic rehabilitation of the edentulous patient. Early stages of jaw atrophy (Class II, III & IV) result in the collapse of the circumoral musculature causing a narrowing of the mouth, loss of lip support, inversion of the lips and contraction of the cheeks. Late changes of jaw atrophy (Class V & VI) result in changes in vertical facial proportion causing a decrease in lower facial height and an increase in chin prominence. These late skeletal changes accentuate the earlier soft tissue facial effects. PMID- 15337182 TI - Intra-oral removal of stones from the hilum of the submandibular gland: report of technique and morbidity. AB - There is increasing evidence to show that the submandibular gland regains function after stone removal and sialoadenectomy may not be the treatment of choice for proximal calculi. A technique of hilar stone removal is described with results and morbidity reported in a series of 55 patients. Stones were retrieved in 54 patients (98%) but four glands (8%) were subsequently removed due to recurrent obstruction. There were no complications related to the procedure (nerve injury, bleeding, infection or scarring) and the morbidity was less than reported for sialoadenectomy. PMID- 15337183 TI - Immunohistochemical localization of cyclooxygenase-1 and -2 in synovial tissues from patients with internal derangement or osteoarthritis of the temporomandibular joint. AB - This study examined the immunohistochemical expression and localization of cyclooxygenase-1 and -2 (COX-1 and COX-2) in synovial tissues from patients with internal derangement (ID) or osteoarthritis (OA) of the temporomandibular joint (TMJ). Synovial tissues from patients with condylar fractures of the mandible were studied as control. Synovial tissues from 13 TMJs of 10 patients with ID or OA and from 5 TMJs of 4 patients with fractures were examined for COX-1 and COX-2 expression by immunohistochemical staining using two monoclonal antibodies. In addition, whether the COX-2 expression grade correlated with the synovitis score and clinical findings was assessed. COX-2 was expressed in the synovial lining, infiltrating mononuclear cells, fibroblast-like cells, and blood vessels, including CD31-positive endothelial cells, in the synovium of patients with ID or OA. Expression levels of COX-1 in synovial lining cells and endothelial cells were similar in the specimens obtained from the patients with ID or OA and those obtained from the controls. The expression of COX-2 positively correlated with arthroscopic findings of synovitis (p = 0.55, P = 0.023) and with joint pain (p = 0.56, P = 0.021). These results suggest that up-regulation of COX-2 in synovium may play a part in the pathogenesis of synovitis in patients with ID or OA of the TMJ. PMID- 15337184 TI - Clinical significance and usefulness of quantification of telomerase activity in oral malignant and nonmalignant lesions. AB - We quantified telomerase activity (TA) in patients with oral and maxillofacial malignant and nonmalignant lesions, and compared it with their clinical status and grade of malignancy. Fifty-two malignant and 52 nonmalignant lesions were analyzed. All malignant lesions were pathologically diagnosed as oral squamous cell carcinoma (OSCC). Normal gingival tissue served as a control. These specimens were obtained by biopsy or surgical resection, and stored at -80 degrees C until use. TA was quantified by a fluorescence-based TRAP method. TA levels ranged from 0.00 to 95.24 (average 33.24)U/microgP in 52 malignant lesions, and from 0.00 to 79.35 (average 11.91)U/microgP in 52 nonmalignant lesions (P < 0.0001). TA was detected in 96.2% of malignant and 65.4% of nonmalignant lesions. There was no relationship between TA levels and clinical stages or YK classification. However, under WHO classification, there were significant differences (P < 0.05) between Grades I and III or II + III. Among nonmalignant lesions, epithelial dysplasia showed a significantly higher TA level than that of oral lichen planus (P < 0.05) and other benign lesions (P < 0.0001). Oral lichen planus also significantly differed from other benign lesions (P < 0.05). These results suggest that TA is related to the histological grade of malignancy, and is also useful as a prognostic predictor for precancerous lesions and conditions. PMID- 15337185 TI - Service provision by patient and visit characteristics in Australian oral and maxillofacial surgery: 1990 to 2000. AB - This study was set-up to describe main areas of service by patient and visit characteristics and compare trends in services between 1990 and 2000. All registered oral and maxillofacial surgeons in Australia were surveyed in 1990 and 2000 using mailed self-complete questionnaires. Service provision data were collected from a one-week log. Data were available from 79 surgeons from 1990 (response = 73.8%) and 116 surgeons from 2000 (response = 65.1%). Service distributions were dominated by dentoalveolar surgery in 1990 (66.6%) and 2000 (63.5%). Multivariate analysis showed: patient age, location of visit (office/theatre/inpatient facility) and referral source (general/specialist and dental/medical) were associated with all five main areas of service; type of visit (consult/operation/review) was associated with four main areas; patient sex and place of visit (private/public) was associated with three main areas; the only significant change over time was an increased percentage of orthognathic surgery, odds ratio = 1.4 (95% CI: 1.1-1.7) times higher in 2000 compared to 1990. Main areas of service were associated with a range of explanatory variables such as age and sex of patients, and place, location and type of visit, and referral source. However, the distribution of services remained relatively stable over time. PMID- 15337186 TI - The Elastic Internal Traction (EIT): an effective method to reduce the displaced facial fractures. AB - Open reduction and rigid fixation are commonly used to treat displaced fractures of the facial skeleton. Manual reduction can be performed by means of forceps or transosseus wires to close the bone fragments. In order to reduce facial fractures, we used a technique, called Elastic Internal Traction (EIT), based on the elastic action of rubber bands stretched between screws placed on both sides of the fracture line. We have used EIT in 104 patients suffering from mandibular fractures, and 40 cases of patients with orbito-maxillary complex fractures out of the 707 patients treated for trauma between July 2000 and August 2002 at our hospital. In our opinion, this technique provides an effective reduction and a stable primary fixation of the bone before the final fixation. It also has the advantage that the surgical field is clear of the assistant's hands and surgical instruments that are usually used to lock the reduction. The resulting operative time is shortened, and the plating of the bone is simplified. PMID- 15337187 TI - Metastatic adenocarcinoma of the temporomandibular joint from the cardia of the stomach: a case report. AB - A case is presented of a 67-year-old man with adenocarcinoma of the gastric cardia that metastasised to the temporomandibular joint (TMJ). A carcinoma of the cardia metastatic to the TMJ has not previously been reported. The case is unusual in many respects. The condyle and disc were anteriorly displaced by the tumour, resulting in progressive mandibular deviation and crossbite. Radiographically, however, no destructive changes of the joint were observable. Tumour staging showed this lesion to be the only distant metastasis. PMID- 15337188 TI - Multiple sclerosis and the visual system. AB - Visual symptoms frequently complicate the course of multiple sclerosis (MS). The ophthalmologist must be familiar with the neuro-ophthalmologic manifestations of MS to facilitate the initial evaluation and treatment of patients. Magnetic Resonance Imaging has become an important tool to confirm the diagnosis of MS or to assess the risk of MS in patients with clinically isolated demyelinating syndromes. This article reviews the neuro-ophthalmologic manifestations of MS and the management issues that arise in early MS. PMID- 15337189 TI - Ocular myasthenia gravis. AB - Myasthenia gravis may be limited solely to the eyes (ocular myasthenia) or may have systemic manifestations (generalized myasthenia). Most patients display blepharoptosis or ophthalmoparesis, resulting in diplopia, but the ocular motility disorder may take many forms. The diagnosis is often elucidated by variable muscle weakness with fatigability noted in the history and on examination. The ice test has supplanted the need for the edrophonium (Tensilon) test in many cases of ocular myasthenia. Therapy may be challenging, because many ocular myasthenics do not achieve significant clinical improvement with acetylcholinesterase inhibitors alone. Immunomodulatory treatment,such as systemic corticosteroids, is frequently effective, and emerging data suggest that this type of therapy may reduce the likelihood of ocular myasthenia becoming generalized. PMID- 15337190 TI - Neuro-ophthalmic manifestations of neurodegenerative disease. AB - Patients who have neurodegenerative disease frequently present to the ophthalmologist with visual complaints. The causes of the symptoms are varied and include visual sensory and ocular motor dysfunction. This article provides an overview of the neuroophthalmic manifestations of the most common neurodegenerative diseases presenting with visual dysfunction, including Alzheimer's disease, Parkinson's disease, progressive supranuclear palsy, and others. Evaluation of visual function in patients who have neurodegenerative disorder can be challenging, but familiarity with the type of visual dysfunction most commonly associated with each disorder will allow for a focused assessment and treatment when possible. PMID- 15337191 TI - Functional visual loss. AB - Functional visual loss is a common problem encountered in practice. It must be recognized that this problem occurs in patients who have organic illness. Manual perimetry is the most effective method for evaluating functional visual loss, and the presence of a central scotoma in a functional visual field strongly suggests that organic pathology is present. Neuropsychological and psychiatric intervention can be useful in treating these patients. PMID- 15337192 TI - Cerebrovascular disease and the visual system. AB - Vascular diseases of the brain affect vision owing to a variety of mechanisms and cause a range of signs and symptoms. These conditions may result from diminished bloodflow, from leakage of blood through vessel walls, or from the compression of adjacent structures by enlarged vessels. Vascular disease may affect the arterial or venous systems and may be caused by abnormal communications between arteries and veins. In some cases, these visual manifestations have a significant impact on quality of life; in other cases, they may be warning signs of serious underlying neurologic or systemic disease. This article emphasizes the conditions that present most commonly to ophthalmologists. PMID- 15337193 TI - The eye and headache. AB - Ophthalmologists are often the first physicians to evaluate patients with headaches, eye pain, and headache-associated visual disturbances. Although ophthalmic causes are sometimes diagnosed, most eye pain and many types of visual disturbances are neurologic in origin. Afferent and efferent symptoms and signs are associated with headache disorders. This article reviews the primary headache disorders and focuses on their ophthalmic manifestations. The major divisions are migraine and the trigeminal autonomic cephalgias. PMID- 15337194 TI - Neuro-ophthalmic manifestations of systemic disease--hematology. AB - The hematopoietic system is a complex mixture of circulating blood cells, plasma, and plasma proteins. Disorders of the hematologic system may result from excess or impaired production of blood cells, destruction of blood cells, or abnormal function of existing blood cells. The afferent and efferent visual systems may both be affected by blood cell disorders, and the specific manifestation is dependent upon which cell line is involved, the type of dysfunction, and the characteristics of the target tissue. This article reviews the ocular and neuro ophthalmic manifestation of hematologic disorders. PMID- 15337195 TI - Neuro-ophthalmologic manifestations of systemic disease: rheumatologic/Inflammatory. AB - Rheumatologic and inflammatory systemic diseases often cause similar neurologic and neuro-ophthalmologic manifestations. A wide variety of conditions can be included in these categories. This article summarizes the most common of these conditions, including sarcoidosis, idiopathic orbital inflammation, inflammatory bowel disease, Wegener's granulomatosis, polyarteritis nodosa, Churg-Strauss syndrome, Behcet's disease, systemic lupus erythematosis, scleroderma, Sjogren's syndrome, and rheumatoid arthritis. PMID- 15337196 TI - Systemic infections of neuro-ophthalmic significance. AB - Systemic infectious disease may present with or include neuro-ophthalmic findings. Many of these signs and symptoms are related to topographical location of the infectious process,but some entities have distinctive and characteristic features that allow specific diagnosis. This article updates and reviews the neuro-ophthalmic features of infectious disease. PMID- 15337197 TI - Endocrine and metabolic deficiency. AB - Endocrine and metabolic disorders that cause neuro-ophthalmologic manifestations are frequently encountered by ophthalmologists in clinical practice. This review focuses on three of the most common entities for which neuro-ophthalmologic signs and symptoms are a prominent feature. These entities include pituitary disorders (adenoma, apoplexy, and hypophysitis), thyroid-associated ophthalmopathy, and neuro-ophthalmologic complications of diabetes mellitus, such as ischemic ocular motor mononeuropathies and diabetic papillopathy. PMID- 15337198 TI - Genetic disorders and the optic nerve: a clinical survey. AB - Genetic disorders can cause a broad range of optic nerve pathology. Clinical symptoms and examination findings of optic nerve dysfunction may provide early clues to the presence of inherited genetic disease. For many disorders, molecular genetic testing is available for the diagnosis of affected individuals and the identification of unaffected carriers. This article surveys a broad array of systemic genetic disorders affecting the optic nerve, reviewing their ocular manifestations, systemic findings, and clinical genetics. PMID- 15337199 TI - Paraneoplastic disorders of ophthalmic interest. AB - Although paraneoplastic syndromes are not common, their recognition is important. Diagnosis of a paraneoplastic disorder can lead to the diagnosis of an underlying tumor or avoid the continued search for metastasis in a patient who has a known cancer. In addition, appropriate diagnosis leads to providing appropriate treatment and prognostic information. PMID- 15337200 TI - Neuro-ophthalmologic diagnosis and therapy of central nervous system trauma. AB - Although direct injury to the orbit and globes can easily result in ophthalmologic disorders, it is less appreciated that distant head injury can similarly result in injury to the retrobulbar afferent visual pathways or cranial nerves. The physician must be suspicious of latent and manifest injuries to the neuro-ophthalmologic system after head trauma and maintain an awareness of the need for timely intervention in a subset of these disorders. Even patients with injuries removed from the eyes should be checked for visual problems, and if any are detected, a careful neuro-ophthalmologic examination should be performed. In most cases, examination and judicious use of neuroimaging will help in choosing the appropriate management for injuries to the afferent and efferent visual systems. PMID- 15337201 TI - Imaging the neurovisual system. AB - This article reviews the neuroimaging modalities commonly used in neuro ophthalmology. These modalities are illustrated with several cases in an atlas format to highlight the differences between each technique. The focus is a review of MRI, MR angiography, CT,CT angiography, and digital subtraction angiography. PMID- 15337202 TI - Optic neuropathies caused by toxins and adverse drug reactions. AB - Toxic optic neuropathy refers to the ingestion of a toxin or an adverse drug reaction that results in vision loss from optic nerve damage. Patients may present with bilateral sudden loss of vision in the setting of an acute intoxication or an insidious asymmetric loss of vision from an adverse drug reaction. Toxins and drugs associated with a toxic optic neuropathy may directly harm the optic nerve; however, some drugs are associated with an ischemic optic neuropathy or optic neuritis, in which optic nerve damage is an indirect effect of the causative agent. The most important aspect of treatment is recognition and drug withdrawal. Patients need to be appropriately counseled, particularly in the setting of amiodarone-associated optic neuropathy. PMID- 15337204 TI - Anemia as a risk factor and therapeutic target in heart failure. AB - Anemia has recently been recognized as an important comorbid condition and potentially novel therapeutic target in patients with heart failure (HF). Anemia is common in HF patients, with a prevalence ranging from 4% to 55% depending on the population studied. Multiple potential mechanisms of interaction exist between anemia and the clinical syndrome of HF, including hemodilution, inflammatory activation, renal insufficiency, and malnutrition. A growing body of literature from observational databases and clinical trials suggests that anemia is an independent risk factor for adverse outcomes in patients with HF. Although preliminary data suggest that treatment of anemia may result in significant symptomatic improvement in HF, aggressive treatment of anemia may also be associated with increased risk of hypertension or thrombosis. Multiple ongoing studies will provide definitive data on the balance of risks and benefits of anemia treatment in chronic HF. PMID- 15337205 TI - Clinical and angiographic outcome after sirolimus-eluting stent implantation in aorto-ostial lesions. AB - OBJECTIVES: This observational study evaluated the clinical and angiographic outcomes of patients with aorto-ostial coronary artery disease treated with sirolimus-eluting stents (SESs) or with bare metal stents (BMSs). BACKGROUND: The safety and effectiveness of SESs for the treatment of aorto-ostial lesions have not been demonstrated. METHODS: We identified 82 consecutive patients who underwent percutaneous coronary interventions in 82 aorto-ostial lesions using the SES (32 patients) or BMS (50 patients) and compared the two groups of patients. The incidence of major adverse cardiac events (MACE), including death or Q-wave myocardial infarction (MI), target lesion revascularization (TLR), and target vessel revascularization (TVR), were recorded in-hospital and at a 10 month follow-up. RESULTS: All stents were implanted successfully. There were no statistically significant differences regarding major in-hospital complications between the two groups. At 10-month follow-up, two (6.3%) patients in the SES group and 14 (28%) patients in the BMS group underwent TLR (p = 0.01); MACE were less frequent in the SES group compared to the BMS group (19% vs. 44%, p = 0.02). Angiographic follow-up showed lower binary restenosis rates (11% vs. 51%, p = 0.001) and smaller late loss (0.21 +/- 0.31 mm vs. 2.06 +/- 1.37 mm, p < 0.0001) in the SES group. CONCLUSIONS: The main finding of our study is that, compared to the BMS, implantation of the SES in aorto-ostial lesions appears safe and effective, with no increase in major in-hospital complications and a significant improvement in restenosis and late event rates at 10-month follow-up. PMID- 15337206 TI - Focal and multi-focal plaque macrophage distributions in patients with acute and stable presentations of coronary artery disease. AB - OBJECTIVES: This study was designed to utilize optical coherence tomography (OCT) images of coronary atherosclerotic plaque macrophages to investigate the relationship between macrophage distributions and clinical syndrome. BACKGROUND: The relative significance of focal macrophage infiltration and generalized coronary inflammation for predicting acute coronary events is a currently a source of considerable controversy in cardiology. Lack of a high-resolution cross sectional imaging modality has limited macrophage evaluation in vivo. METHODS: Intracoronary OCT imaging was performed at culprit and non-culprit plaques in patients presenting with stable angina pectoris, unstable angina pectoris,and ST segment elevation myocardial infarction. Macrophage densities were quantified from these images and analyzed with respect to the clinical presentations of the patients under investigation. RESULTS: A significantly greater macrophage density was found in unstable patients, both for fibrous and lipid-rich plaques (p = 0.025 and p = 0.002, respectively). Within each patient, the macrophage densities at culprit and non-culprit lesions correlated significantly (r = 0.66, y = 0.88x + 0.43, p = 0.01). Sites of plaque rupture demonstrated a greater macrophage density than non-ruptured sites (6.95 +/- 1.60%, 5.29 +/- 1.17%; p = 0.002). Surface macrophage infiltration was a stronger predictor of unstable clinical presentation than subsurface infiltration for culprit lesions (p = 0.035) but not for remote lesions (p = 0.80). CONCLUSIONS: Our results demonstrate that increases in both multi-focal and focal macrophage densities are highly correlated with symptom severity. By providing a means of detecting increases in plaque macrophage content before an acute event, this technique may aid in determining prognosis and guiding preventive therapy. PMID- 15337207 TI - Association of duration of symptoms at presentation with angiographic and clinical outcomes after fibrinolytic therapy in patients with ST-segment elevation myocardial infarction. AB - OBJECTIVES: We sought to determine if an underlying mechanism of the association between prolonged symptom-to-treatment times and adverse outcomes may be an association of symptom-to-treatment times with impaired Thrombolysis In Myocardial Infarction myocardial perfusion grades (TMPGs). BACKGROUND: Prolonged symptom duration among ST-segment elevation myocardial infarction (STEMI) patients undergoing fibrinolytic therapy is associated with adverse outcomes. METHODS: Angiography was performed 60 min after fibrinolytic administration in 3,845 Thrombolysis In Myocardial Infarction (TIMI) trial patients. RESULTS: The median time from symptom onset to treatment was longer among patients with impaired myocardial perfusion (3.0 h for TMPG 0/1 vs. 2.7 h for TMPG 2/3; p = 0.001). In a multivariate model, impaired tissue perfusion (TMPG 0/1) remained associated with increased time to treatment (odds ratio 1.14 per hour of delay; p = 0.007) even after adjusting for Thrombolysis In Myocardial Infarction flow grade (TFG) 3, left anterior descending infarct location, and baseline clinical characteristics. Impaired myocardial perfusion after rescue/adjunctive percutaneous coronary intervention (PCI) was associated with longer median times to treatment (3.0 h for TMPG 2/3 vs. 2.7 h for TMPG 0/1; p = 0.017), as was abnormal epicardial flow after rescue/adjunctive PCI (3.3 h for TFG 0/1/2 vs. 2.8 h for TFG 3; p = 0.005). Thirty-day mortality was associated with longer time from onset of symptoms to treatment (6.6% mortality for time to treatment >4 h vs. 3.3%; p < 0.001), even among patients undergoing rescue PCI. CONCLUSIONS: A prolonged symptom to treatment time among STEMI patients is associated with impaired myocardial perfusion independent of epicardial flow both immediately after fibrinolytic administration and after rescue/adjunctive PCI. These data provide a pathophysiologic link between prolonged symptoms due to vessel occlusion, impaired myocardial perfusion, and poor clinical outcomes. PMID- 15337208 TI - Cardiac rehabilitation after myocardial infarction in the community. AB - OBJECTIVES: The aim of this study was to examine participation in cardiac rehabilitation after myocardial infarction (MI) by age and gender and the association of participation with survival. BACKGROUND: Lesser participation in cardiac rehabilitation has been reported for women and the elderly. METHODS: All incident MIs in Olmsted County were validated. Baseline characteristics and outcomes were ascertained from the medical record. Logistic regression examined the association between participation, age, and gender. Propensity scores were used to examine the association between participation and outcome. RESULTS: Among 1,821 persons with incident MI (58% men, 46% age >70 years), 55% participated in cardiac rehabilitation. Participants were more likely to be men, younger, and have fewer comorbidities (p < 0.01 for all comparisons). After adjustment, women were 55% less likely to participate than men (odds ratio [OR] 0.45, 95% confidence interval [CI] 0.34 to 0.60), and persons 70 years or older were 77% less likely to participate than persons younger than 60 (OR 0.23, 95% CI 0.16 to 0.33). Participants had a lower risk of death and recurrent MI at three years (p < 0.001 and p = 0.049, respectively). The survival benefit associated with participation was stronger in more recent years (relative risk [RR] for 1998 vs. 1982 0.28, 95% CI 0.18 to 0.43; RR for 1990 vs. 1982 0.41, 95% CI 0.33 to 0.52). CONCLUSIONS: Approximately half of the patients participated in cardiac rehabilitation after MI. Participation did not increase over time. Women and elderly persons were less likely to participate, independently of other characteristics. Participation in rehabilitation was independently associated with decreased mortality and recurrent MI, and its protective effect was stronger in more recent years. PMID- 15337209 TI - Type D personality predicts death or myocardial infarction after bare metal stent or sirolimus-eluting stent implantation: a Rapamycin-Eluting Stent Evaluated at Rotterdam Cardiology Hospital (RESEARCH) registry substudy. AB - OBJECTIVES: We investigated the effect of Type D personality on the occurrence of adverse events at nine months in patients with ischemic heart disease (IHD) after percutaneous coronary intervention (PCI) with sirolimus-eluting stents (SESs) or bare stents. Type D patients experience increased negative emotions and tend not to express these emotions in social interactions. BACKGROUND: The SES is a new advent in interventional cardiology that reduces the restenosis rate and the risk of a major adverse cardiac event, but the SES has not been shown to confer any benefits on death or myocardial infarction (MI). METHODS: Consecutive patients with IHD (n = 875) enrolled in the Rapamycin-Eluting Stent Evaluated At Rotterdam Cardiology Hospital (RESEARCH) registry completed the Type D Personality Scale (DS14) six months after PCI. The end point was a composite of death and MI. Events occurring before administration of the DS14 were excluded from analyses. RESULTS: At nine months' follow-up, there were 20 events. Type D patients were at a cumulative increased risk of adverse outcome compared with non-Type D patients: 5.6% versus 1.3% (p < 0.002). Type D personality (odds ratio [OR] 5.31; 95% confidence interval [CI] 2.06 to 13.66) remained an independent predictor of adverse outcome adjusting for all other variables, including SES versus bare stent implantation. CONCLUSIONS: Type D personality was an independent predictor of adverse events in patients optimally treated with the latest advent in interventional cardiology. The DS14 could be used as a screening instrument in routine clinical practice to optimize risk stratification in IHD patients. PMID- 15337210 TI - Low cholesterol, mortality, and quality of life in old age during a 39-year follow-up. AB - OBJECTIVES: We assessed the impact of serum cholesterol level in early midlife on total mortality during up to 39 years of follow-up and on the quality of life (QoL) in old age. BACKGROUND: Total effects of low serum cholesterol on health have been in dispute, especially in elderly persons, and there are few data on the long-term effects of low cholesterol on QoL. METHODS: The cohort consisted of 3,277 healthy businessmen age 30 to 45 years at baseline (1960s). In addition to baseline, serum cholesterol values were available for part of the cohort in 1974, 1986, and 2000. The QoL was assessed in 80.9% of survivors (n = 1,820, mean age 73 years) with a RAND-36 (SF-36) QoL questionnaire in 2000. Mortality up to 2002 (n = 1,173) was retrieved from national registers. RESULTS: Cholesterol was clearly reduced in survivors during follow-up, except in the lowest baseline serum cholesterol group. Baseline cholesterol predicted 39-year total mortality in a graded manner (p < 0.0001), and a value < or =5.0 mmol/l was associated with a 25% reduction in total mortality. In old age, the physical component summary score of RAND-36 was significantly (p = 0.02) higher (better) in the lowest baseline cholesterol group; no difference was found in the mental component summary score (p = 0.51). CONCLUSIONS: Low serum cholesterol level in midlife predicted not only better survival but also better physical function and QoL in old age, without adversely affecting mental QoL. PMID- 15337211 TI - Low and lowered cholesterol and total mortality. PMID- 15337212 TI - African Americans and Caucasians have a similar prevalence of coronary calcium in the Dallas Heart Study. AB - OBJECTIVES: We sought to compare the prevalence of coronary atherosclerosis in a cohort of middle-age African American (black) and non-Hispanic Caucasian (white) men and women from a population-based probability sample. BACKGROUND: Blacks have a higher mortality from coronary heart disease (CHD) than whites, particularly among younger individuals, and yet several studies have reported that coronary atherosclerosis is less prevalent in blacks than in whites. Data from population based samples comparing coronary atherosclerotic burden between blacks and whites are limited. METHODS: The prevalence of coronary atherosclerosis in middle-aged blacks and whites was determined using coronary calcium measured by electron beam computed tomography in 1,289 men and women from a population-based probability sample from Dallas, Texas. RESULTS: The population estimates of the frequency of a positive scan for coronary artery calcium were not statistically different between black and white men (37% vs. 41%, p = 0.36) or between black and white women (29% vs. 23%, p = 0.21). Although the prevalence of most of the coronary risk factors varied significantly between blacks and whites, mean Framingham coronary risk factor scores were identical in black and white men (10 +/- 4) but significantly higher in black women (13 +/- 4) than in white women (12 +/- 4). CONCLUSIONS: Blacks in the general population have a prevalence of coronary atherosclerosis similar to whites. Factors other than coronary atherosclerotic burden, which are not reflected in the Framingham risk score, contribute significantly to the higher CHD mortality rate in blacks. PMID- 15337213 TI - Atherosclerosis of the aorta: risk factor, risk marker, or innocent bystander? A prospective population-based transesophageal echocardiography study. AB - OBJECTIVES: The goal of this study was to investigate whether complex aortic atherosclerosis is associated with increased risk of vascular events in a non selected population. BACKGROUND: In selected high-risk patients, aortic atherosclerosis is associated with increased risk of vascular events. METHODS: We describe the relationship between simple versus complex (>4-mm thick or mobile debris) aortic atherosclerotic plaques and vascular events during follow-up in a random sample of 585 persons (age > or =45 years) using 1993 to 2000 data from the Stroke Prevention: Assessment of Risk in a Community (SPARC), a prospective population-based longitudinal study. RESULTS: At five-year median follow-up (range, 0.5 to 6.5 years), cardiac events (death, non-fatal myocardial infarction, coronary revascularization, heart failure associated with coronary artery disease) and cerebrovascular events (ischemic fatal and non-fatal strokes, transient ischemic attacks) had occurred in 95 subjects and 41 subjects, respectively. Age, male gender, prior coronary artery disease, higher pulse pressure, and diabetes were significant cardiovascular predictors. Age, prior myocardial infarction, and a history of atrial fibrillation were significant cerebrovascular predictors. Simple aortic plaques (253 persons) were not independently associated with either cardiac or cerebrovascular events. Complex plaques (44 persons) were marginally associated with cardiac events, adjusting for age and gender (hazard ratio [HR], 2.28; 95% confidence interval [CI], 1.11 to 4.68; p = 0.053 for two degrees of freedom [complex and simple plaques vs. no plaques]) but not after adjusting for additional clinical risk factors (HR, 1.22; 95% CI, 0.57 to 2.62; p = 0.64). Complex plaques were associated with cerebrovascular events only univariately. CONCLUSIONS: Aortic atherosclerotic plaques are not associated with future cardiac or cerebrovascular events. Aortic atherosclerosis may not be an independent risk factor for vascular events in the general population. PMID- 15337214 TI - Predictors of mortality in patients with heart failure and preserved systolic function in the Digitalis Investigation Group trial. AB - OBJECTIVES: We identified predictors of mortality in patients with preserved ejection fraction (EF) and clinical heart failure (HF). BACKGROUND: Although diastolic HF is common, the factors that predict mortality have not been clearly defined. METHODS: We studied 988 patients with HF and preserved EF enrolled in the Digitalis Investigation Group (DIG) trial. Survival analyses were employed to identify variables associated with mortality. RESULTS: During 3.1 years of follow up, 231 (23%) patients died. Among 18 variables considered, the strongest independent predictors of death were glomerular filtration rate (adjusted hazard ratio for one standard deviation decrease 1.50, 95% confidence interval [CI] 1.35 to 1.67, p < 0.0001), New York Heart Association functional class III or IV (adjusted hazard ratio 1.64, 95% CI 1.20 to 2.18, p = 0.0011), male gender (adjusted hazard ratio 1.71, 95% CI 1.26 to 2.32, p = 0.0005), and older age (adjusted hazard ratio for one standard deviation increase of age2 1.28, 95% CI 1.08 to 1.50, p = 0.0019). A risk score was developed to estimate long-term mortality. CONCLUSIONS: Diastolic HF is associated with a high death rate. Important predictors of death include impaired renal function, worse functional class, male gender, and older age. PMID- 15337215 TI - Accurate and reproducible measurement of left ventricular volume and ejection fraction by contrast echocardiography: a comparison with magnetic resonance imaging. AB - OBJECTIVES: We evaluated the accuracy and reproducibility of contrast echocardiography versus tissue harmonic imaging for measurements of left ventricular (LV) volumes and ejection fraction (EF) compared to magnetic resonance imaging (MRI). METHODS: Digital echo recordings of apical LV views before and after intravenous contrast were collected from 110 consecutive patients. Magnetic resonance imaging of multiple short-axis LV sections was performed with a 1.5-T scanner. Left ventricular volumes and EF were calculated offline by method of discs. Thirty randomly selected patients were reanalyzed for intraobserver and interobserver variability. RESULTS: Compared with baseline, contrast echo increased feasibility for single-plane and biplane volume analysis from 87% to 100% and from 79% to 95%, respectively. The Bland-Altman analysis demonstrated volume underestimation by echo, but much less pronounced with contrast. Limits of agreement between echo and MRI narrowed significantly with contrast: from -18.1% to 8.3% to -7.7% to 4.1% (EF), from -98.2 to -11.7 ml to 59.0 to 10.7 ml (end-diastolic volume), and from -58.8 to 21.8 ml to -38.6 to 23.9 ml (end-systolic volume). Ejection fraction from precontrast echo and MRI differed by > or =10% (EF units) in 23 patients versus 0 after contrast (p < 0.001). At intraobserver and interobserver analysis, limits of agreement for EF narrowed significantly with contrast. CONCLUSIONS: The two-dimensional echocardiographic evaluation of LV volumes and EF in non-selected cardiac patients was found to be more accurate and reproducible when adding an intravenous contrast agent. PMID- 15337216 TI - Detection of retained microbubbles in carotid arteries with real-time low mechanical index imaging in the setting of endothelial dysfunction. AB - OBJECTIVES: We sought to determine if intravenously injected microbubbles would be retained by the carotid arteries (CAs) in the setting of endothelial dysfunction (ED) using a linear transducer equipped with a low mechanical index pulse sequence scheme (PSS). BACKGROUND: Microbubbles normally pass freely through large and small vessels but are retained in regions with ED. New high frequency low mechanical index PSS can potentially be utilized to image these retained microbubbles. METHODS: Intravenous albumin- and lipid-encapsulated microbubbles were administered in seven pigs while imaging the CAs before and after a 20% intralipid infusion to induce hypertriglyceridemia. The degree of microbubble retention was quantified by measuring endothelial acoustic intensity (AI) after clearance of free-flowing microbubbles. Microbubble adherence was also evaluated after selective balloon injury of the CAs. The CA diameter responses to acetylcholine were quantified. RESULTS: After induction of hypertriglyceridemia, adherence of albumin-encapsulated microbubbles was visually evident in all CAs, and endothelial AI increased significantly (p < 0.001 compared with baseline). The CA responses to acetylcholine went from vasodilation at baseline to vasoconstriction during hypertriglyceridemia. Endothelial AI also increased in the balloon-stretched vessels (p < 0.01 compared with uninjured vessels) after albumin-encapsulated microbubble injection, with a ring of microbubbles selectively adhering to the injured segment. This retention was not observed with lipid-encapsulated microbubbles. Scanning electron microscopy confirmed that albumin-coated microbubbles adhered to endothelial cells. CONCLUSIONS: Retention of intravenously injected albumin microbubbles occurs in the setting of both global and regional ED in large vessels and can be noninvasively imaged with high frequency low mechanical index PSS. PMID- 15337217 TI - Prognostic role of B-type natriuretic peptide levels in patients with type 2 diabetes mellitus. AB - OBJECTIVES: We hypothesized that B-type natriuretic peptide (BNP) levels can predict cardiac mortality in diabetic patients. BACKGROUND: Detection of cardiovascular disease in diabetics can be difficult until overt events occur. METHODS: A total of 482 diabetics (majority male with type 2 diabetes) at the Veterans Affairs Medical Center San Diego were divided into two groups: 1) referred patients for echocardiogram on the basis of clinical suspicion of cardiac dysfunction (referred [R], n = 180); 2) patients randomly selected from the diabetic clinic without any suspicion of cardiac dysfunction (not referred [N R], n = 302). We examined cardiac events and all-cause mortality in relation to initial BNP levels during the follow-up. RESULTS: A total of 71 (14.7%) patients died during this period: 52 of 180 (29%) in the R group (30 of 52 [58%] cardiac, 10 of 52 [19%] non-cardiac, 2 of 52 [4%] renal, 10 of 52 [19%] unknown cause) and 19 of 302 (6%) in N-R group (6 of 19 [32%] cardiac). The median BNP level in the R and N-R groups who died of cardiac, non-cardiac, and unknown cause was 537 and 87, 80 and 53, and 343 and 38 pg/ml, respectively. The receiver-operating characteristic (ROC) values for mortality in two groups in relation to BNP revealed the area under the curve to be 0.720 and 0.691, respectively (p < 0.01 in both). Among commonly used prognostic indicators in diabetics, only the ROC for triglycerides was significant. The most accurate cut-point in both the N-R group (87%) and R group (61%) was 120 pg/ml of BNP. Cox regression analysis showed BNP to be the most significant predictor of all-cause mortality in the R group. There was a marked decrease in survival in the patient group with BNP >120 pg/ml. CONCLUSIONS: B-type natriuretic peptide appears to be a reliable predictor of future cardiac and all-cause mortality in diabetic patients. PMID- 15337218 TI - Impact of physical deconditioning on ventricular tachyarrhythmias in trained athletes. AB - OBJECTIVES: The purpose of this research was to evaluate the impact of athletic training and, in particular, physical deconditioning, on frequent and/or complex ventricular tachyarrhythmias assessed by 24-h ambulatory (Holter) electrocardiogram (ECG). BACKGROUND: Sudden deaths in athletes are usually mediated by ventricular tachyarrhythmias. METHODS: Twenty-four hour ambulatory ECGs were recorded at peak training and after a deconditioning period of 19 +/- 6 weeks (range, 12 to 24 weeks) in a population of 70 trained athletes selected on the basis of frequent and/or complex ventricular tachyarrhythmias (i.e., > or =2,000 premature ventricular depolarization [PVD] and/or > or =1 burst of non sustained ventricular tachycardia [NSVT]/24 h). RESULTS: A significant decrease in the frequency and complexity of ventricular arrhythmias was evident after deconditioning: PVDs/24 h: 10,611 +/- 10,078 to 2,165 +/- 4,877 (80% reduction; p < 0.001) and NSVT/24 h: 6 +/- 22 to 0.5 +/- 2, (90% reduction; p = 0.04). In 50 of the 70 athletes (71%), ventricular arrhythmias decreased substantially after detraining (to <500 PVDs/24 h and no NSVT). Most of these athletes with reduced arrhythmias did not have structural cardiovascular abnormalities (37 of 50; 74%). Over the 8 +/- 4-year follow-up period, each of the 70 athletes survived without cardiac symptoms. CONCLUSIONS: Frequent and/or complex ventricular tachyarrhythmias in trained athletes (with and without cardiovascular abnormalities) are sensitive to brief periods of deconditioning. In athletes with heart disease, the resolution of such arrhythmias with detraining may represent a mechanism by which risk for sudden death is reduced. Conversely, in athletes without cardiovascular abnormalities, reduction in frequency of ventricular tachyarrhythmias and the absence of cardiac events in the follow-up support the benign clinical nature of these rhythm disturbances as another expression of athlete's heart. PMID- 15337219 TI - Athens, athletes, and arrhythmias: the cardiologist's dilemma. PMID- 15337220 TI - Prognostic implications of atrial fibrillation in patients undergoing myocardial perfusion single-photon emission computed tomography. AB - OBJECTIVES: The aim of this research was to determine whether presence of atrial fibrillation (AF) provides incremental prognostic information relative to myocardial perfusion single-photon emission computed tomography (MPS) with respect to risk of cardiac death (CD). BACKGROUND: The prognostic significance of AF in patients undergoing MPS is not known. METHODS: A total of 16,048 consecutive patients undergoing MPS were followed-up for a mean of 2.21 +/- 1.15 years for the development of CD. Of those, 384 patients (2.4%) had AF. Cox proportional hazards method was used to compare clinical and perfusion data for the prediction of CD in patients with and without AF. RESULTS: Atrial fibrillation was a significant predictor of CD in patients with normal (1.6% per year vs. 0.4% per year in non-AF patients), mildly abnormal (6.3% per year vs. 1.2% per year), and severely abnormal MPS (6.4% per year vs. 3.7% per year) (p < 0.001 for all). By multivariable analysis, AF patients had worse survival (p = 0.001) even after adjustment for the variables most predictive of CD: age, diabetes, shortness of breath, use of vasodilator stress, rest heart rate, and the nuclear variables. In the 4,239 patients with left ventricular ejection fraction evaluated by gated MPS, AF demonstrated incremental prognostic value not only over clinical and nuclear variables, but also over left ventricular ejection in predicting CD (p = 0.014). CONCLUSIONS: The presence of AF independently increases the risk of cardiac events over perfusion and function variables in patients undergoing MPS. Patients with AF have a high risk of CD, even when MPS is only mildly abnormal. Whether patients with AF and mildly abnormal MPS constitute a group more deserving of early referral to cardiac catheterization is a question warranting further study. PMID- 15337221 TI - Left atrial tachycardia after circumferential pulmonary vein ablation for atrial fibrillation: electroanatomic characterization and treatment. AB - OBJECTIVES: The purpose of this study was to evaluate the electroanatomic characteristics of left atrial tachycardia (AT) in a series of patients who underwent circumferential pulmonary vein ablation (CPVA) and to describe the ablation strategy and clinical outcome. BACKGROUND: Circumferential pulmonary vein ablation is an effective treatment for atrial fibrillation. A potential midterm complication is the development of left AT. There are only isolated reports describing mapping and ablation of such arrhythmias. METHODS: Thirteen patients (age 57.4 +/- 8.9 years, five female) underwent mapping and ablation of 14 left ATs via an electroanatomic mapping system a mean of 2.6 +/- 1.6 months after CPVA. RESULTS: Three patients were characterized as having focal AT (cycle length: 266 +/- 35.9 ms). Of 11 macro-re-entrant tachycardias studied in the remaining 10 patients (cycle length: 275 +/- 75 ms), 5 showed single-loop and 6 dual-loop circuits. Re-entrant circuits used the mitral isthmus, the posterior wall, or gaps on previous encircling lines. Such gaps and all three foci occurred anterior to the left superior pulmonary vein or at the septal aspect of the right pulmonary veins. Thirteen of 14 tachycardias (93%) were successfully ablated. CONCLUSIONS: Left AT after CPVA can be due to a macro-re-entrant or focal mechanism. Re-entry occurs most commonly across the mitral isthmus, the posterior wall, or gaps on previous ablation lines. Such gaps and foci occur most commonly at the anterior aspect of the left superior pulmonary vein and at the septal aspect of the right pulmonary veins. These arrhythmias can be successfully mapped and ablated with an electroanatomic mapping system. PMID- 15337222 TI - Non-contact mapping to guide radiofrequency ablation of atypical right atrial flutter. AB - OBJECTIVES: This study was aimed at evaluating the efficacy of non-contact mapping and ablation of non-incisional atypical right atrial (RA) flutters. BACKGROUND: The majority of atypical RA flutters were reported in patients after surgical incision of the RA. METHODS: The study group consisted of 15 patients (61 +/- 13 years, 8 males) with atypical atrial flutter (AFL). The RA activation during AFL was delineated using a non-contact mapping system (EnSite 3000 with Precision Software, Endocardial Solutions, St. Paul, Minnesota). The narrowest part of each reentrant circuit was targeted using radiofrequency energy. RESULTS: In all 15 patients, non-contact mapping showed AFLs confined to the RA with RA activation time accounting for 100% of the cycle length (210 +/- 19 ms). During single-loop re-entry in seven patients, the activation wave front circulated around the central obstacle (CO) in the anterolateral wall with conduction through the channel between the CO and the crista terminalis (CT). During figure of-eight re-entry in eight patients, simultaneous upper and lower loop re-entry through the conduction gap in the CT was found in four patients, and simultaneous upper loop and free-wall single-loop re-entry was observed in four patients. Radiofrequency ablation of the free-wall channel and/or CT gap was effective in eliminating these AFLs in 13 patients. During a follow-up of 16.8 +/- 3.8 months, two patients had recurrence of left AFL, and one had recurrence of atrial fibrillation. CONCLUSIONS: Atypical RA flutters could arise from single-loop or double-loop figure-of-eight re-entry. Radiofrequency ablation of the free-wall channel and/or the CT gap was effective in eliminating these arrhythmias. PMID- 15337223 TI - Doppler-guided regulation of a telemetrically operated adjustable pulmonary banding system. AB - OBJECTIVES: We report on the Doppler-assessed regulation of an adjustable pulmonary artery band (PAB) in an animal model and in our first group of patients. BACKGROUND: Indications for pulmonary artery banding have expanded to include patients requiring a late arterial switch. A telemetry-operated, fully implantable, adjustable PAB system (FloWatch- PAB, Endoart SA, Lausanne, Switzerland) has been developed to facilitate these operations. METHODS: The device was implanted in 13 minipigs (age one to five months, weights 3.2 to 12.0 kg). The main study was performed on nine minipigs with adjustments of the PAB at implantation and at 1, 3, 5, 8, and 12 weeks after, assessed by Doppler pressure gradients. Explanation was performed 12 weeks after surgery. A long-term histology study (6 months and 14 months after surgery) was done on the other four minipigs. After approval by the ethics committee, the device was implanted in eight patients with weights between 2.8 and 9 kg to decrease pulmonary blood flow and pressure and to retrain the left ventricle before arterial switch. The device was progressively tightened, with increasing transband Doppler gradients. Follow up was one to three months. RESULTS: An excellent correlation between transbanding systolic pressure gradient and degree of PAB constriction was encountered in the minipig study as well as in the human setting. No early or late deaths or reoperations occurred. Malfunction of the device was noted in three of 21 implanted devices. Two were related to surgically inflicted damage at implantation and one to an electronic problem that was fixed by resetting the control device. CONCLUSIONS: The device offers a Doppler-controllable adjustment of pulmonary blood flow. It permits controlled tightening and release of the band, which improves perioperative and postoperative courses and decreases surgical interventions to adjust tightness of the band. It allows a protracted occlusion protocol, which may provide the best effect on retraining the left ventricle. PMID- 15337224 TI - Predictors of sudden cardiac death after Mustard or Senning repair for transposition of the great arteries. AB - OBJECTIVES: The goal of this research was to identify predictors for sudden death (SD) in patients with transposition of the great arteries (TGA) who have undergone atrial inflow repair. BACKGROUND: Sudden death is the most common cause of late death after atrial inflow repair of TGA. Little is known about the predictors of SD. METHODS: This was a retrospective, multicenter, case-controlled study. We identified 47 patients after Mustard's or Senning's operation who experienced an SD event (34 SD, 13 near-miss SD). Each patient was matched with two controls with the same operation, but without an SD event. Information on numerous variables before the event was obtained and compared with controls at the same time frame. RESULTS: Presence of symptoms of arrhythmia or heart failure at most recent follow-up and history of documented arrhythmia (atrial flutter [AFL]/atrial fibrillation [AF]) were found to increase the risk of SD. Electrocardiogram (ECG), chest X-ray, and Holter ECG findings were not predictive of SD. Neither medication nor pacing was found to be protective. Most SD events (81%) occurred during exercise. Ventricular tachycardia/ventricular fibrillation were the recorded rhythm during SD in 21 of 47 patients. CONCLUSIONS: Presence of symptoms and documented AFL/AF are the best predictors of SD in TGA patients. Patients with these findings should be further evaluated for risk of SD. PMID- 15337225 TI - Multiple, brief coronary occlusions during early reperfusion protect rabbit hearts by targeting cell signaling pathways. AB - OBJECTIVES: An in situ model was used to test whether and how multiple occlusions at reperfusion can protect rabbit myocardium. BACKGROUND: Recently it was demonstrated that postconditioning in dogs salvaged ischemic myocardium. METHODS: Control hearts underwent 30-min regional ischemia/3-h reperfusion, whereas in experimental hearts four postconditioning cycles of 30-s occlusion/30-s reperfusion starting 30 s after release of the index coronary occlusion were added in the presence or absence of various cell signaling antagonists. RESULTS: Postconditioning decreased infarction from 35.4 +/- 2.7% of the risk zone in control hearts to 19.8 +/- 1.8% (p < 0.05). Six cycles did not result in greater protection. If postconditioning cycles were begun after 10 min of reperfusion, protection was no longer evident. Either the non-selective K(ATP) channel closer glibenclamide or the putatively selective mitochondrial K(ATP) channel antagonist 5-hydroxydecanoate administered 5 min before reperfusion blocked the protection afforded by postconditioning, indicating involvement of the mitochondrial K(ATP) channel. PD98059, a mitogen-activated protein/extracellular-signal regulated kinase (MEK) 1/2 and therefore extracellular-signal regulated kinase (ERK) inhibitor, and N(omega)-nitro-L-arginine methyl ester, an antagonist of nitric oxide synthase, infused shortly before reperfusion also aborted the protection afforded by postconditioning. Combined ischemic postconditioning and preconditioning resulted in significantly greater protection than either alone. CONCLUSIONS: Multiple, short, regional coronary occlusions immediately after prolonged myocardial ischemia are an effective cardioprotective intervention in the rabbit, and the mechanism of protection involves activation of ERK, production of nitric oxide, and opening of mitochondrial K(ATP) channels. These observations suggest that a similar approach could be applied in the cardiac catheterization laboratory to protect reperfused myocardium after primary angioplasty in patients with acute myocardial infarction. PMID- 15337226 TI - Postconditioning: old wine in a new bottle? PMID- 15337227 TI - Fibroblast growth factor-1 improves cardiac functional recovery and enhances cell survival after ischemia and reperfusion: a fibroblast growth factor receptor, protein kinase C, and tyrosine kinase-dependent mechanism. AB - OBJECTIVES: We sought to investigate the role of fibroblast growth factor (FGF)-1 during acute myocardial ischemia and reperfusion. BACKGROUND: The FGFs display cardioprotective effects during ischemia and reperfusion. METHODS: We investigated FGF-1-induced cardioprotection during ischemia and reperfusion and the intracellular signaling pathways responsible for these effects in an ex vivo murine setup of myocardial ischemia and reperfusion. RESULTS: Cardiac-specific human FGF-1 overexpression was associated with enhanced post-ischemic hemodynamic recovery and decreased lactate dehydrogenase release during reperfusion. Inhibition of the FGF receptor, protein kinase C (PKC), and tyrosine kinase (TK) resulted in blockade of FGF-1-induced protective effects on cardiac functional recovery and cell death. CONCLUSIONS: The overexpression of FGF-1 induces cardioprotection through a pathway that involves the FGF receptor, PKC, and TK. PMID- 15337228 TI - Myocardial gene transfer by selective pressure-regulated retroinfusion of coronary veins: comparison with surgical and percutaneous intramyocardial gene delivery. AB - OBJECTIVES: We sought to study adenoviral gene delivery using percutaneous selective pressure-regulated retroinfusion and to compare it directly with surgical and percutaneous intramyocardial delivery (PIMD) for the first time. BACKGROUND: Intramyocardial delivery (IMD) has been recommended to be the preferred gene delivery strategy so far. However, surgical and percutaneous intramyocardial injection lead to incomplete retention of the injected viral vectors and to limited spatial myocardial distribution. Percutaneous selective pressure-regulated retroinfusion of the coronary veins was developed recently to provide an effective and more homogenous regional myocardial gene transfer. METHODS: In 15 pigs, adenoviral vectors (Ad2-CMV beta-galactosidase [beta-gal] 5 x 10(9) pfu) were applied via surgical IMD (n = 5), PIMD (n = 5), and selective pressure-regulated retroinfusion (n = 5). Seven days after gene transfer, myocardial beta-gal expression was measured by ELISA. RESULTS: Selective retroinfusion into the anterior cardiac vein substantially increased reporter gene expression (1,039 +/- 79 pg beta-gal/mg protein) in the targeted left anterior descending coronary artery territory when compared with surgical (448 +/ 127, p < 0.05) and PIMD (842 +/- 145, p < 0.05). Both IMD approaches showed an inhomogenous beta-gal expression, particularly along the injection sites, while retroinfusion resulted in a more homogenous transmural gene expression. CONCLUSIONS: Percutaneous selective pressure-regulated retroinfusion compares favorably with surgical and percutaneous intramyocardial injection techniques by providing a more homogenous and even more efficient adenoviral gene delivery. PMID- 15337229 TI - The year in health care delivery and outcomes research. PMID- 15337230 TI - President's page: Cardiology without borders: embracing a healthy global association. PMID- 15337232 TI - Do cypher gene mutations cause left ventricular noncompaction with subclinical myopathy? PMID- 15337233 TI - Severe ostial saphenous vein graft disease leading to acute coronary syndromes after proximal aorto-saphenous anastomoses with the symmetry bypass connector device. PMID- 15337235 TI - Optimal target international normalized ratio for patients with mechanical heart valves. PMID- 15337236 TI - The optimal intensity of vitamin K antagonists in patients with mechanical heart valves: a meta-analysis. PMID- 15337237 TI - Anticoagulation management of patients with prosthetic valves. PMID- 15337240 TI - Tibolone: how does its mechanism of action translate into clinical effects. PMID- 15337239 TI - ACC/AHA 2004 guideline update for coronary artery bypass graft surgery: summary article. A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee to Update the 1999 Guidelines for Coronary Artery Bypass Graft Surgery). PMID- 15337241 TI - Tibolone: a selective tissue estrogenic activity regulator (STEAR). PMID- 15337242 TI - The molecular determinants of estrogen receptor pharmacology. AB - The classical models of steroid receptor pharmacology held that agonists functioned by binding to their cognate receptors, facilitating their conversion from an inactive form to one that was capable of activating transcription. By extrapolation, it was believed that antagonists functioned by competitively inhibiting agonist binding, freezing the receptor in an inactive state. However, as early as 1967 when the biologic actions of the "antiestrogen" tamoxifen were first described, it was clear that this simple model did not adequately describe estrogen receptor (ER) pharmacology. Since these initial observations, significant progress has been made in defining the molecular mechanism(s) by which cells distinguish between different ER ligands. The most important of these are: (i) differences in the relative expression level of the two ER subtypes, alpha and beta, in target tissues; (ii) the impact, which the bound ligand has on ER structure; and (iii) the complement of coactivators and corepressors in a target cell, which can interact with the activated receptor. This review describes how these advances have impacted our understanding of the pharmacologic activities of currently available ER ligands. PMID- 15337243 TI - Cellular homeostasis and the breast. AB - Increasing our knowledge regarding the control of cellular homeostasis in the normal human breast is important in understanding how breast tumours arise and whether compounds used for hormone replacement therapy are able to promote tumour formation In our studies on tissue from pre-menopausal, non-pregnant, non lactating women, oestradiol is the main ovarian steroid mitogen for the breast epithelium whereas progesterone has little or no effect. Oestradiol appears to influence proliferative activity indirectly via oestrogen-receptor positive cells which control the activity of adjacent division competent cells by means of paracrine or juxtacrine growth factors. After the menopause, however, a mitogenic effect of progesterone becomes apparent which may be due to the reduction in endogenous oestradiol levels or, possibly, to tissue specific alterations in steroid sensitivity. Whatever the mechanism, the proliferative effects of progesterone on the post-menopausal breast have implications for the use of HRT. PMID- 15337244 TI - Role of enzymes and tissue-specific actions of steroids. AB - Enzymes, such as steroid sulphatase, sulphotransferases and 17 beta hydroxydehydrogenases, are important in regulating the tissue-specific actions of steroids. These enzymes can exist in different isoforms in different tissues and can be regulated by tissue-specific promoters and factors such as cytokines, prostaglandins and growth factors. The activity of these enzymes and the mechanisms by which they are regulated may help to account for some of the tissue specific effects of the hormone replacement therapy drug tibolone, which has oestrogenic effects on the bone but not on the breast or endometrium. PMID- 15337245 TI - Multisystem evaluations of the long-term effects of tibolone on postmenopausal monkeys. AB - This long-term study (2 years) was designed to compare the effects of tibolone (LoTib at 0.05 mg/kg and HiTib at 0.2 mg/kg) with those of conjugated equine oestrogens (CEE) alone (0.042 mg/kg) and CEE continuously combined with medroxyprogesterone acetate (MPA) (0.167 mg/kg) on coronary artery atherosclerosis, bone, mammary gland and uterus in ovariectomised cynomolgus monkeys fed a moderately atherogenic diet. Despite reductions in plasma concentrations of high density lipoprotein cholesterol in tibolone-treated monkeys, there was no exacerbation of coronary artery atherosclerosis. Tibolone was equivalent to, or slightly better than, CEE and CEE + MPA in protecting against postmenopausal bone loss and loss of bone strength. Tibolone also resulted in less stimulation of breast and endometrial tissue compared with CEE and CEE + MPA. In conclusion, the results suggest that tibolone is a cardiovascular-safe treatment that is effective for the prevention of osteoporosis and that may have advantages over CEE or CEE + MPA with regard to endometrial and breast safety. PMID- 15337246 TI - Tissue-selectivity: the mechanism of action of tibolone. AB - Tibolone is effective in preventing bone loss and treating climacteric symptoms, without stimulating the endometrium. The effects on bone, brain and vagina can be accurately explained by the oestrogenic activity of tibolone, but oestrogenic activity is not expressed in the endometrium. Tibolone behaves differently from oestrogen plus progestogen combinations on the breast. Therefore, tibolone can be characterised as a selective oestrogen activity regulator. The objective of this review is to characterise the typical properties of tibolone in order to explain its tissue-selective action. Tibolone is rapidly converted into three major metabolites: 3 alpha- and 3 beta-hydroxy-tibolone, which have oestrogenic effects, and the Delta(4)-isomer, which has progestogenic and androgenic effects. The 3-hydroxy metabolites are present in the circulation, predominantly in their inactive sulphated form. The tissue-selective effects of tibolone are the result of metabolism, enzyme regulation and receptor activation that vary in different tissues. The bone preserving effects are the result of oestradiol receptor activation, whilst other steroid receptors, notably the progesterone and androgen receptor, are not involved. Breast tissue of monkeys is not stimulated, as occurs with oestrogen plus progestogen, because tibolone and its metabolites inhibit sulphatase and 17 beta-hydroxysteroid dehydrogenase (HSD) type I and stimulate sulphotransferase and 17 beta-HSD type II, the combined effects of which prevent conversion to active oestrogens. In addition, tibolone affects cellular homeostasis in the breast by inhibiting proliferation and stimulating apoptosis. Tibolone does not stimulate the endometrium because of the action of the highly stable progestogenic metabolite (Delta(4)-isomer) in combination with an effect on the sulphatase (inhibition)-sulphotransferase (stimulation) system. The oestrogenic metabolites of tibolone have direct favourable effects on the cardiovascular system and, in in vivo models, tibolone has shown no adverse consequences. In conclusion, tibolone shows oestrogenic effects in brain, vagina and bone and has direct oestrogenic effects on the cardiovascular system. In the endometrium, the progestogenic activity of the Delta(4)-metabolite and the effect on oestrogen-inactivating enzymes prevent oestrogenic stimulation. The mammary gland is not stimulated in currently used animal models. Tibolone appears to regulate estrogenic activity in the various tissues by influencing the availability of estrogenic compounds for the estradiol receptor in a tissue selective manner. PMID- 15337247 TI - New findings on X-linked Adrenoleukodystrophy: 5alpha-reductase isoform 2 relative gene expression is modified in affected fibroblasts. AB - X-linked Adrenoleukodystrophy (X-ALD) is a neurodegenerative disease with an endocrinological component since, in addition to the nervous system, the adrenal cortex and the testis are mainly affected, with corresponding clinical signs. 5Alpha-reductase, a key enzyme in steroid hormone metabolism, catalyzes the conversion of testosterone into the potent androgen dihydrotestosterone and other metabolic steps in steroidogenesis. It is present in two isoforms, 5alpha reductase isoform 1 and 2, that are encoded by different genes. The isoforms are differently expressed in the tissues, where they have distinct physiological relevance. Our study shows that the expression of isoform 2, evaluated by Real Time PCR, is significantly altered in fibroblasts from patients affected by X-ALD with respect to controls, whereas isoform 1 is not affected. This is the first demonstration of an alteration of 5alpha-reductase isoform 2 gene expression in X ALD, that may be related to the steroidogenesis impairment and to the specific organ malfunction. PMID- 15337248 TI - Localization of DJ-1 mRNA in the mouse brain. AB - DJ-1 is mutated in autosomal recessive, early onset Parkinson's disease but the exact localization of the DJ-1 gene product in the mammalian brain is largely unknown. We aimed to evaluate the DJ-1 mRNA expression pattern in the mouse brain. Serial coronal sections of brains of five male and five female adult mice were investigated by using in situ hybridization with a DJ-1 specific 35S-labeled oligonucleotide probe. Hybridized sections were analyzed after exposure to autoradiography films and after coating with a photographic emulsion. DJ-1 was heterogeneously expressed throughout the mouse central nervous system. A high expression of DJ-1 mRNA was detected in neuronal and non-neuronal populations of several structures of the motor system such as the substantia nigra, the red nucleus, the caudate putamen, the globus pallidus, and the deep nuclei of the cerebellum. Furthermore, DJ-1 mRNA was also highly expressed in non-motor structures including the hippocampus, the olfactory bulb, the reticular nucleus of the thalamus, and the piriform cortex. The high expression of DJ-1 mRNA in brain regions involved in motor control is compatible with the occurrence of parkinsonian symptoms after DJ-1 mutations. However, expression in other regions indicates that a dysfunction of DJ-1 may contribute to additional clinical features in patients with a DJ-1 mutation. PMID- 15337249 TI - Lesions of the periaqueductal gray block the medial preoptic area-induced activation of the urethrogenital reflex in male rats. AB - The medial preoptic area (MPOA) is important for male sexual behavior, including erections and ejaculation. Stimulation of the MPOA evokes urethrogenital reflex like responses. However, the descending pathways mediating this response are unknown. We examined the effect of bilateral lesions of the periaqueductal gray (PAG) on the MPOA-induced response. Electrical stimulation of the MPOA was used to induce rhythmic motor patterns of the bulbospongiosus muscle, discrete regions of the PAG were lesioned and the response to MPOA stimulation re-examined. These studies demonstrate that the descending pathway mediating the MPOA-induced activation of the urethrogenital reflex-like responses travel through and may relay in the PAG. PMID- 15337250 TI - Possible involvement of the facial nucleus in regulation of respiration in rats. AB - The facial nucleus (FN) has been known as a motor nucleus to control the activity of the facial skeletal muscles by its efferent somatic motoneurons. Much less, however, is known about the non-motor control functions of its interneurons. The present study was designed to investigate if the interneurons of the FN participate in controlling rhythmic respiration in the sodium thiopental anesthetized and vagotomized Sprague-Dawley rats with facial motoneurons retrogradely degenerated with techniques of electrical and chemical stimulation of the FN and extracellular recording of discharge of neurons in the FN. Single pulse stimulation (75-100 microA, 0.2 ms) of the FN during inspiration caused a transient restrain in phrenic discharge. Short train stimulation (75-100 microA, 0.2 ms, 100 Hz, 3-5 pulses) delivered during the early- or mid-term of inspiration augmented the inspiratory duration, but switched the inspiration off when delivered during the later stage of inspiration. Short train stimulation delivered during expiration prolonged the expiratory duration. Continuous stimulation could inhibit the inspiration. Microinjection of kainic acid into the FN caused an augmentation in inspiratory duration and amplitude and in expiratory duration. These data indicate that the interneurons of the FN might participate in the modulation of respiration. Different discharge patterns of interneurons in the FN, interestingly some respiratory related patterns, were observed, which provide a possible structural basis for the role of the FN in respiratory regulation. PMID- 15337251 TI - Experimental herpes simplex virus encephalitis: a long-term study of interleukin 6 expression in mouse brain tissue. AB - This study aimed to investigate the expression of interleukin-6 (IL-6) in acute and chronic herpes simplex virus encephalitis. In the brain of 15 SJL mice infected with herpes simplex virus type 1, strain F, and 14 control animals we performed a sequential quantitative analysis of expression of IL-6 mRNA with reverse transcription real-time polymerase chain reaction. The viral burden peaked in the acute disease, and then returned to a low baseline value. At day 7 following infection, IL-6 expression was significantly (2.05-fold) increased as compared with the baseline expression in uninfected animals. Twenty-one days after infection the mRNA expression still was significantly (1.78-fold) upregulated. No significant differences of IL-6 mRNA expression between infected and control mice were found after 2 and 6 months. We observed a 2.5-fold increase of IL-6 mRNA expression in control mice with increasing age of animals. We have additionally studied the clinical evolution of HSVE in IL-6 deficient mice. In experimental herpes simplex virus encephalitis IL-6, as a potent mediator of neuronal injury, is upregulated in the acute but not in the chronic disease. IL-6 deficient mice presented early and severe clinical signs of HSVE as compared to the wild-type C57/bl6 mice. PMID- 15337252 TI - Selective reduction of chromogranin A-like immunoreactivities in the prefrontal cortex of schizophrenic subjects: a postmortem study. AB - It is suggested that secretogranins/chromogranins play a role in regulating secretion of various proteins and amines, including neurotransmitters from secretory granules. Several studies have implicated the importance of altered synaptic connectivity in schizophrenia. We employed immunohistochemical techniques to determine if the level of chromogranin A (CgA)-immunoreactivity (IR) was altered in the subjects with schizophrenia. Nine subjects with schizophrenia and nine age- and sex-matched control subjects were selected for this study. Immunohistochemistry using specific antibody against CgA was performed on sections of prefrontal cortex and hippocampus. Images of CgA-IR were analyzed by computer-based image analyzing software. CgA-IR was significantly decreased in layers III-V of the prefrontal cortex in schizophrenic subjects compared with control subjects. In the hippocampus, no significant difference was observed between two groups. The results indicate that there may be a decrease in the number of CgA positive large dense-core vesicles per terminal, and/or in the number of CgA positive terminals, suggesting possible functional impairment of prefrontal synaptic contact in schizophrenia. PMID- 15337253 TI - Impairment of a cortical event-related desynchronisation during a bimanual load lifting task in children with autistic disorder. AB - In autism, the abilities of communication are affected, associated with abnormalities of cognitive, sensorial and motor development. In a previous study based on a load-lifting task, we showed impairment of anticipation in children with autism as evidenced by kinematics and eletromyographic recordings [Neurosci. Lett. 348 (2003) 17]. In the present study, we assessed the cortical counterparts of the use of anticipatory postural adjustments in a group of control children and in a group of children with autism. The tasks required maintaining a stable forearm position despite imposed or voluntary lifting of an object placed either on the controlateral forearm or on a support. We investigated the differences between the two groups of children on the Event-Related Desynchronisation (ERD) which precedes movement onset in adults [Electroencephalogr. Clin. Neurophysiol. 46 (1979) 138]. Electroencephalogram (EEG) power evolution of a 6-8-Hz frequency band was averaged before and after imposed or voluntary movement onset. EEG reactivity of control and autistic children did not differ during the imposed unloading condition, but significant differences appeared in the voluntary unloading situations. Before lifting the object, control children showed an ERD above the left motor areas. An ERD also occurred above the right motor areas when the object was placed on their forearm. This indicates that the ERD can also translate the use of anticipatory postural adjustments. By contrast, children with autism did not show an ERD in the two voluntary situations. This suggests a central deficit of anticipation in both postural and motor control in children with autism. PMID- 15337254 TI - The impact of adenosine on the release of acetylcholine, dopamine, and norepinephrine from the cat carotid body. AB - Exogenously administered adenosine provokes an increase in respiration in both animal models and in man. Administered near the carotid body adenosine increases neural output from the carotid body in rats and cats. Hypoxia has the same effect. Hypoxia also provokes a release of acetylcholine (ACh), dopamine (DA), and norepinephrine (NE) from the carotid body. The present study aimed to determine the effect of exogenous adenosine on the release of ACh, DA, and NE from the carotid bodies of cats. After a recovery period (from surgery) carotid bodies were first incubated for 10 (DA, NE) or 15 (ACh) min in Eppendorf tubes containing 85 microL of a physiological salt solution equilibrated with 40% O2/5% CO2 at 37 degrees C (hyperoxia). At the end of the incubation period the medium was drawn off, and measured for ACh, DA, and NE using HPLC-ECD methods. Next 85 microL of the medium and the tubes were equilibrated with a hypoxic gas mixture (4% O2/5% CO2) and the carotid bodies were incubated for 10 (DA, NE) or 15 (ACh) min, at the end of which the medium was drawn off and measured for ACh, DA, and NE. In the ACh studies there followed a post-hypoxic hyperoxic exposure (40% O2/5% CO2). ACh tubes were then made 100 microM with respect to adenosine, and the hyperoxic, hypoxic, and post-hypoxic hyperoxic challenges were repeated. One of the two DA, NE tubes had the 100 microM adenosine from the start. Adenosine significantly increased the release of ACh, but significantly decreased the hypoxia-induced release of DA. Potential mechanisms for these changes are reviewed. PMID- 15337255 TI - Sociability trait and serotonin metabolism in the rat social interaction test. AB - Social behaviour is the basis of one of the most generally accepted independent dimensions of personality. The purpose of the present study was to find out whether the social activity of individual rats, expressed in the social interaction test of anxiety, is consistent, and associated with monoamine levels. Four social interaction tests with 10 days intervals were carried out in 20 rats, and the animals were decapitated 4 days after the last test. There was no consistent correlation between performances in single tests, but the social interaction time in each test correlated strongly with the mean values of social activity in all or the other three tests. Social interaction time of rats correlated moderately but significantly with their partner's social activity in the test. The average social interaction time correlated strongly with 5-HIAA levels in the frontal cortex (r = -0.67, P < 0.01). Neither exposure of rats singly to the social interaction test box nor the test procedure had any effect on monoamine levels. When animals were decapitated immediately after a single social interaction test, there was a negative correlation between the social interaction time and 5-HIAA and 5-HT levels in the septum, but not in the frontal cortex or hippocampus. Thus, social behaviour is a stable trait, expression of which depends in part upon the partner's social behaviour. This trait is negatively associated with 5-HT metabolism in the frontal cortex. Social activity of rats in a particular test situation may rather be related to 5-HT metabolism in the septum. PMID- 15337256 TI - Distribution of 5-HT7 receptors in the human brain: a preliminary autoradiographic study using [3H]SB-269970. AB - The distribution of the 5-HT7 receptor was analyzed using the selective 5-HT7 receptor antagonist radioligand [3H]SB-269970 and human brain whole hemisphere autoradiography. The results indicated that 5-HT7 receptors are most abundantly localized in the anterior thalamus and in the dentate gyrus. Other regions containing intermediate levels of 5-HT7 receptors included the hypothalamus, anterior cingulate gyrus, hippocampus, amygdala and certain brainstem nuclei. The distribution pattern obtained in this study supports the involvement of this receptor subtype in affective behavior and cognition. PMID- 15337257 TI - Systematic scaling of target width: dynamics, planning, and feedback. AB - The target width of a single target in a two-target reciprocal aiming task was scaled from small (ID = 5.85) to large (ID = 2.85) and large-to-small within individual trials with movement amplitude fixed. Scaling target width produced a transition in the end-effector's dynamics and based on a measure of movement harmonicity, the transition was sensitive to the initial conditions but not to the direction of target width scaling. Hysteresis emerged in a variety of kinematic measures suggesting that the interdependency of planning and feedback control processes was sensitive to initial conditions as well as the direction of target width scaling. Practice increased the efficiency of the reciprocal movements and produced changes in movement time and the measure of harmonic motion that revealed a tuning of the end-effector's dynamics to cyclical motion over as large of range of IDs as possible. The tuning occurred through the modulation of time spent accelerating and decelerating the end-effector for IDs outside the range of 3.85-4.26. The results are discussed with reference to a critical ID boundary that separates regions of parameter space wherein the end effector's dynamics are more cyclical (limit-cycle) or discrete (fixed-point) in nature. PMID- 15337258 TI - Effect of Vim thalamic DBS in Parkinson's disease on F wave duration. AB - F waves were recorded from abductor hallucis muscle in eight Parkinsonian patients with deep brain stimulation (DBS) electrodes surgically implanted in their Vim thalamic nucleus in two conditions of DBS ON and OFF. Patients with relatively anteriorly located electrodes exhibited a significant reduction in F wave duration and also in the UPDRS rigidity score of the corresponding foot when the DBS was ON. In contrast, patients with relatively posteriorly located electrodes exhibited no significant difference in F wave duration in the two DBS ON and OFF conditions. The rigidity UPDRS score in the corresponding foot diminished very little in the latter group. Both groups had great improvement in their tremor at rest UPDRS score in that foot when the DBS was ON. Vim surgery is generally accepted to affect tremor mechanisms. However, surgical intervention in anterior parts of Vim has been reported to affect rigidity mechanisms. This correspondence of these two symptoms of rigidity and tremor with the two locations of anterior and relatively posterior Vim may indicate the contribution of mechanisms of rigidity, but not tremor, in enhancement of F wave duration and hyper excitability of spinal motoneuron. PMID- 15337259 TI - Treatment with subthreshold doses of caffeine plus trihexyphenidyl fully restores locomotion and exploratory activity in reserpinized rats. AB - Trihexyphenidyl (THP) is a drug commonly used to reduce parkinsonian symptoms. An important side effect of this agent is memory impairment. Since caffeine enhances the potency of THP to inhibit haloperidol-induced catalepsy, caffeine may be used as an adjuvant of lower doses of THP, in order to improve its antiparkinsonian effects without causing memory disruption. To further assess the synergism between caffeine and THP, both drugs were tested in reserpinized rats, another preclinical model of Parkinson's disease. Four groups of rats (n = 7) were treated with reserpine (5 mg/kg, i.p.). A control group (n = 7) was treated only with the vehicle for reserpine (dimethylsulphoxide). The spontaneous locomotor behavior was tested 24 h later in a box with infrared sensors, 30 min after receiving one of the following treatments: distilled water (1 ml/kg), caffeine (1 mg/kg), THP (0.1 mg/kg) or caffeine plus THP. The levels of horizontal locomotion (14 +/- 5%) and vertical exploration (15 +/- 10%) were significantly lower in reserpinized rats treated with distilled water, compared with the mean activity values (100%) recorded in animals pretreated only with the vehicle for reserpine. The reserpine-induced hypokinesia was neither reversed by caffeine alone nor by THP alone. However, the combination of caffeine plus THP restored locomotion (141 +/- 19%) and vertical exploration (82 +/- 17%) to levels not significantly different to those of non-reserpinized rats. Moreover, the time-course of locomotion and exploration displayed the characteristic habituation over time, in which short-term memory processes are involved. Also, the thigmotaxis index indicated that the combined treatment did not induce anxiety-like behavior. Hence, these results support the proposal that low, subthreshold doses of caffeine plus THP have the potential to alleviate the motor disabilities in parkinsonian patients, with a low risk of causing anxiety or memory impairment. PMID- 15337260 TI - Recording of ventral posterior lateral thalamus neuron response to contact heat evoked potential in patient with neurogenic pain. AB - Microrecording of single unit response to contact heat-evoked potential (CHEP) were performed in right ventral posterior lateral (VPL) thalamus during deep brain stimulation (DBS) surgery in a patient with chronic neurogenic pain. In our patient, neurons (n = 10) recorded in the ventral thalamus fired at a higher rate of 40 Hz compared to neurons recorded in Parkinsonian patients (24 Hz). Contact heat was applied by a fast heating and cooling probe of 5 cm2 area on the dermatome C6 territory of the left hand. One out of four thalamic cells located in the VPL responded repetitively 325 ms after the peak temperature was reached with a burst of action potential, suggesting A-delta fibre activation. This observation supports the use of CHEP for mapping nociceptive neurons location during DBS surgery for intractable pain. PMID- 15337261 TI - Interaction between matrix metalloproteinase 3 and the epsilon4 allele of apolipoprotein E increases the risk of Alzheimer's disease in Finns. AB - Polymorphisms affecting the expression of matrix metalloproteinases (MMPs), i.e. proteolytic enzymes that degrade intercellular material, have been found at position -1607 (1G/2G) in MMP1 and at -1171 (5A/6A) in MMP3. Interestingly, elevated levels of MMP1 and MMP3 have been observed in the brains of Alzheimer's disease (AD) patients and those of tissue inhibitors of MMPs in the cerebrospinal fluid of AD and Parkinson's disease (PD) patients, suggesting a role for MMPs in these disorders. The aim was to investigate a possible association between the afore-mentioned MMP1 and MMP3 polymorphisms and the risk of developing AD or PD. The polymorphisms were genotyped in 97 AD, 52 PD and 101 control patients. We found an interaction between MMP3*5A and APOE 4 alleles (P < 0.0001) which increases the risk of AD (OR: 23.7, 95% CI: 5.8-144.9, P < 0.0001) compared to those who possess neither MMP3*5A nor APOE 4. In conclusion, our finding suggests that the MMP3 gene, especially together with APOE 4, may contribute to the development of AD. PMID- 15337262 TI - Inflammatory glial activation in the brain of a patient with hereditary sensory neuropathy type 1 with deafness and dementia. AB - The brain of a patient with hereditary sensory neuropathy type 1 (HSN-1) associated with sensorineural deafness and early-onset dementia was neuropathologically investigated. Widespread neuronal degeneration in cerebral neocortex, hippocampus and basal ganglia was revealed, accounting for the clinical features. Loss of neurons with ballooning of residual neurons was remarkable in the hippocampus and frontal, parietal, and occipital lobes. Neuronal degeneration in these regions was accompanied by axonal dystrophy and glial reactions such as microgliosis and astrocytosis, however, only glial responses were prominent in the basal ganglia, brain-stem and cerebellum with mild neuronal loss. These results indicate that the widespread neuronal degeneration may be accelerated by inflammatory processes including glial activation in the brain of a patient with HSN-1 associated with deafness and dementia. PMID- 15337263 TI - Effects of nitric oxide on dentate gyrus cell proliferation after seizures induced by pentylenetrazol in the adult rat brain. AB - Epileptic seizures have been shown to increase the proliferation of granule cell precursors in the adult brain, but the underlying mechanisms remain largely unknown. This study examined the effect of nitric oxide (NO) on the proliferation of granule cell precursors in adult rats after pentylenetrazol (PTZ)-induced generalized clonic seizures. Using systemic bromodeoxyuridine (BrdU) to label dividing cells, we found that injection of the neuronal nitric oxide synthase (nNOS) inhibitor 7-nitroindazole (50 mg/kg i.p.) 10 min before PTZ significantly reduced the number of BrdU labeled cells in the dentate gyrus 3, 7, and 14 days after seizures (P < 0.05). Administration of the inducible NOS (iNOS) inhibitor aminoguanidine (100 mg/kg i.p.) also significantly inhibited the proliferation rate of neural precursor cells in the dentate gyrus at various time points after PTZ-induced seizures. Our findings suggest that epileptic seizures lead to increased cell proliferation in the adult rat dentate gyrus through NO-dependent mechanisms. Both the NO originating from nNOS and iNOS may be involved in brain repair after seizures. PMID- 15337264 TI - Microglial activation is a pharmacologically specific marker for the neurotoxic amphetamines. AB - Neurotoxic amphetamines cause damage to monoamine nerve terminals of the striatum by unknown mechanisms. Microglial activation contributes to the neuronal damage that accompanies injury, disease, and inflammation, but a role for these cells in amphetamine-induced neurotoxicity has received little attention. We show presently that D-methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), D amphetamine, and p-chloroamphetamine, each of which has been linked to dopamine (DA) or serotonin nerve terminal damage, result in microglial activation in the striatum. The non-neurotoxic amphetamines l-methamphetamine, fenfluramine, and DOI do not have this effect. All drugs that cause microglial activation also increase expression of glial fibrillary acidic protein (GFAP). At a minimum, microglial activation serves as a pharmacologically specific marker for striatal nerve terminal damage resulting only from those amphetamines that exert neurotoxicity. Because microglia are known to produce many of the reactive species (e.g., nitric oxide, superoxide, cytokines) that mediate the neurotoxicity of the amphetamine-class of drugs, their activation could represent an early and essential event in the neurotoxic cascade associated with high-dose amphetamine intoxication. PMID- 15337265 TI - Indirubin-3'-oxime inhibits c-Jun NH2-terminal kinase: anti-apoptotic effect in cerebellar granule neurons. AB - Previous studies have demonstrated that c-Jun NH2-terminal protein kinase (JNK) plays a crucial role in neuronal apoptosis. Here, we report that indirubin-3' oxime, a known effective inhibitor of cyclin-dependent kinases (CDKs) and glycogen synthase kinase 3-beta (GSK-3beta), has a significant inhibitory effect on JNK. Kinase assay showed that indirubin-3'-oxime directly inhibited the activity of all three isoforms of JNK (JNK1, and JNK3) in vitro, with half inhibition dose (IC50) of 0.8 microM, 1.4 microM, and 1.0 microM, respectively. In cerebellar granule neurons (CGNs), indirubin-3'-oxime blocked c-Jun phosphorylation induced by potassium withdrawal and prevented CGNs from apoptosis in a dose dependent manner. However, inhibitors of CDKs and GSK-3beta were ineffective in reducing c-Jun phosphorylation both in vitro and in vivo, suggesting that indirubin-3'-oxime prevents c-Jun phosphorylation independent of its inhibition on CDKs and GSK-3beta. Our studies give further supports for JNK targeting strategy in preventing neuronal apoptosis. PMID- 15337266 TI - New mutation (T1232P) of the ATP-7B gene associated with neurologic and neuropsychiatric dominance onset of Wilson's disease in three unrelated Colombian kindred. AB - Wilson's disease is an autosomal recessive disorder of hepatic copper metabolism caused by mutations in a gene encoding a copper-transporting P-type ATPase. We report the clinical and molecular characterization of six members from three unrelated Colombian kindred. Completed sequence DNA analysis linked to the gene ATP-7B from patient wd-1 revealed a novel A to C transversion in exon 17 at position 3856 (A3856C) of the ATP-7B mRNA resulting in a threonine for proline substitution at position 1232 of the ATP-7B protein (T1232P). Additionally, two novel polymorphisms were detected (2785G:Gly875 in exon 11; and intron at +38 a > c:tgcgcccga in exon 19). All affected individuals were homozygous for the T1232P mutation and displayed neurologic and neuropsychiatric dominant onset. This work expands the knowledge about the number, type, and implication of mutations in WD. PMID- 15337267 TI - Time window and pharmacological characterisation of kainate-mediated preconditioning in organotypic rat hippocampal slice cultures. AB - Tolerance to normally neurotoxic insults can be induced by prior a preconditioning exposure to a sublethal insult. Kainate toxicity can be attenuated by prior exposure to very low concentrations of kainate both in vivo and in vitro. Using organotypic hippocampal slice cultures from rats we have shown that 5 microM kainate induces a selective lesion in the CA3 region and this can be significantly attenuated by 1 microM kainate administered 1-5 days earlier. The time window for this effect was affected by the length of time in culture, and preconditioning was blocked by NBQX but not the selective AMPA receptor antagonist GYKI53655. These data demonstrate a role for kainate receptors in preconditioning for the first time and show that organotypic cultures can be used as a model to investigate long-term preconditioning mechanisms. PMID- 15337268 TI - Effect of voluntary contraction intensity on the H-reflex and V-wave responses. AB - This study examined the evolution of H-reflex and V-wave responses of soleus muscle during maximal voluntary plantar-flexor contraction. We also investigated the relationship between the V response and force level and between V-wave during maximal voluntary contraction (MVC) and the maximal H reflex at rest. The H reflex and the V-wave responses are measures of motoneuron excitability and also reflect the magnitude of presynaptic inhibition on Ia afferents and the magnitude of descending motor drive. Both may be influenced by postsynaptic inhibition. Twenty male subjects participated in the study and were assigned to one of two groups. The maximal M wave (Mmax) was evoked at rest in the 20 subjects, who then performed 10 maximal voluntary contraction. During MCV performance, a stimulus was delivered at supra-maximal intensity, which allowed us to record the superimposed M wave (Msup) and V wave of the soleus muscle. These parameters were also recorded during sub-maximal contractions (20, 40, 60, 80% of one MVC) in 10 subjects. The maximal H reflex (Hmax), was evoked at rest in the other 10 subjects. These subjects then performed 10 MVC and the Hsup (superimposed H, evoked by means of stimulus at Hmax intensity) was recorded. The results show that the amplitude of maximal M wave increased during MVC (gain 44.52 +/- 10.71%). No significant difference between Hmax/Mmax at rest and the Hsup/Msup ratios during MVC was observed, while an effect of force level on the V/Msup ratio was found. V/Msup and Hmax/Mmax were linearly correlated (r2 = 0.81), but V/Msup was significantly lower (P < 0.01) than Hmax/Mmax. In conclusion, the present study shows that maximal voluntary contractions potentiate some reflex responses. The V wave, which reflects motoneuron excitability presynaptic inhibition of Ia afferents and the magnitude of descending central motor drive to spinal motoneurons, may be a relatively simple method to analyse the modulation adaptive neural alterations at spinal and supraspinal level during voluntary contractions. PMID- 15337269 TI - The intensity dependence of the auditory evoked N1 component as a predictor of response to Citalopram treatment in patients with major depression. AB - The intensity dependence of the auditory evoked N1 ERP component (IDAP) has been suggested as an indicator of central serotonergic neurotransmission with relevance to pharmacological treatment. We report the results of a study evaluating the IDAP in 16 in-patients fulfilling DSM-IV criteria for major depressive episode in the course of treatment with the SSRI Citalopram. Our data revealed a significant correlation between the intensity slopes of the N1 amplitude prior to Citalopram treatment and treatment response: patients with higher intensity slopes of N1 amplitude showed a significantly stronger decrease of HDRS-Score after Citalopram treatment than patients within the lower intensity slope ranges. Our results indicate an association of N1 amplitude intensity dependence with response to antidepressant treatment with Citalopram. PMID- 15337270 TI - Brain-derived neurotrophic factor, apolipoprotein E genetic variants and cognitive performance in Alzheimer's disease. AB - Since greater attention has been paid to the direct link of genetic variation to cognition and memory performance, apolipoprotein E (ApoE) and brain-derived neurotrophic factor (BDNF) have been the two most frequently studied genes. To investigate the effect of BDNF and ApoE polymorphisms on the cognitive profile of mild-moderate Alzheimer's disease (AD) cases, AD patients, genotyped for ApoE and BDNF polymorphisms, underwent extensive neuropsychological investigation. The effect of either ApoE epsilon4 allele and BDNF genetic variant on the neuropsychological pattern of mental impairment was examined both in terms of group differences in performance on the neuropsychological tests between carriers and non-carriers of each variant and by selecting the best predictor of cognitive performance among demographic and genetic factors by means of a multiple regression analysis. Our data confirm a specific effect caused by the presence and amount of ApoE epsilon4 allele, while they suggest that BDNF genetic variants are not a susceptibility factor to AD. PMID- 15337271 TI - The cytoarchitecture of sulcal folding in Heschl's sulcus and the temporal cortex in the normal brain and schizophrenia: lamina thickness and cell density. AB - Developmental and psychiatric disorders, including schizophrenia, may be associated with altered cortical thickness and folding. Two studies were performed: (1) to assess cortical layering around a sulcus; cortical thickness, relative thickness of the supragranular (I-III):infragranular (IV-VI) layers, and cell density were assessed at anatomically defined points around Heschl's sulcus in tissue from 10 controls and 10 schizophrenia patients. (2) To sample sulci of contrasting prominence; sulcal depth, width, lamina thickness, and cell density from laminae II-VI were taken from various sulci within the temporal lobes from another group of 6 controls and 10 patients. Reduced cell density was found in the fundi of sulci in schizophrenia. Independent of diagnosis; increased sulcal prominence in temporal cortex accompanies reduced lamina thickness (particularly layers V and VI), deep layers show negative relationships between cell density and layer thickness, and total cortex width in Heschl's sulcus reduces by half at the bottom compared to the top. Furthermore, compared to the supragranular layers, the infragranular division is relatively thicker at the top of a gyrus, equal in the wall of the sulcus and relatively thinner at the bottom. Many effects of sulcal folding on laminar proportions in controls are similar in schizophrenia. However, cell density is less at the bottom of some sulci in the temporal lobe in schizophrenia. Sampling methods should consider that cortical folding affects cell and lamina distribution in the sampled region in a highly localised manner. PMID- 15337272 TI - Cyclosporin A and deltamethrin block the downregulation of Nav1.8 sodium channels expressed in Xenopus oocytes. AB - The Nav1.8 sodium channel isoform, expressed in sensory neurons and implicated in pain responses, is known to be upregulated in Xenopus oocytes by agents that activate protein kinase A. In the absence of exogenous modulators, Nav1.8 channels expressed in oocytes exhibited spontaneous downregulation, so that the amplitudes of peak sodium currents at the end of a 30-min recording period were reduced to 58% of those at the outset of recording with no change in the properties of the expressed channels. Perfusion of oocytes with either cyclosporin A or deltamethrin, considered to be diagnostic inhibitors of the protein phosphatase calcineurin, at 10 microM blocked spontaneous downregulation. These results identify endogenous calcineurin as the mediator of Nav1.8 sodium channel downregulation in oocytes. The use of a calcineurin inhibitor such as cyclosporin A provides an effective means of stabilizing the expression of Nav1.8 sodium channels in oocytes for functional and pharmacological studies. PMID- 15337273 TI - Thalamocortical projection from the ventral posteromedial nucleus sends its collaterals to layer I of the primary somatosensory cortex in rat. AB - Here we examined quantitatively axonal projections originating from the ventral posteromedial thalamic nucleus (VPM) to layer I of the primary somatosensory cortex (SI) by extracellular and intracellular injections of biocytin as an anterograde tracer. Following the extracellular injections, two types of VPM afferents with different arborization patterns in SI were observed. The type I extended vertically, forming dense plexus in layers IV and VI, and projected collaterals to layer I. The type II rarely branched in SI, converged in the plexus formed by the type I, and projected no collaterals to the supragranular layers. The labeled fibers in layer I derived from the first type ran parallel to the brain surface, and their mean length was 339.7 +/- 87.5 microm. Intracellular injection into VPM neurons bearing both types of afferent demonstrated the full axonal arborization in both the reticular thalamic nucleus (Rt) and SI. The total length of the axon of a neuron bearing the type I was 86,968.8 microm, and the length of axonal collaterals in layer I of SI was 433.1 microm. The total axonal length of a neuron bearing the type II was very small. The present study is the first to demonstrate substantial projections from VPM to layer I of SI, and provide quantitative data on the entire extent of the axonal arborization of thalamocortical projections from single VPM neurons. PMID- 15337274 TI - Subcellular localization of WD40 repeat 1 protein in PC12 rat pheochromocytoma cells. AB - The dynamics of actin filament protein is crucial for various physiological processes of the cells. Among the proteins correlating with actin dynamics, a novel 67-kDa WD40 repeat protein 1 (WDR1) was the vertebrate homologue of actin interacting protein 1 (Aip1). Even though previous studies have provided the clues on the function of WDR1 in specific organs under pathological conditions, the exact subcellular localization of WDR1 is not known. Therefore, in the present study, we undertook to determine the distribution of WDR1 within PC12 pheochromocytoma cells (PC12 cells) using light and electron microscopic techniques. Double immunocytochemistry clearly showed that WDR1 immunoreactivities (IRs) were co-localized with anti-actin antibody, suggesting the involvement of WDR1 in actin dynamics. WDR1 immunoreactivities (IRs) in PC12 cells showed different distribution patterns as nerve growth factor (NGF) concentrations varied. During active proliferation, the distribution of WDR1 IRs seemed to be similar to those found in cortical actin patches, whereas WDR1 IR was observed in cytoplasmic actin cables after PC12 cells were induced to differentiate by treating with NGF. Though further studies are necessary to determine the function of WDR1, the current data represents a first step towards the in vitro study of WDR1 protein. PMID- 15337276 TI - Politics, normal birth and midwifery. PMID- 15337275 TI - Myosin Va is proteolysed in rat cerebellar granule neurons after excitotoxic injury. AB - Cerebellar granule neurons when exposed to glutamate die through an excitotoxic mechanism induced by overactivation of glutamate receptors. This kind of cell death is mediated by an overload of intracellular calcium involving calpain activation, a Ca2+ -dependent intracellular cysteine protease, among other intracellular responses. On the other hand, class V myosins are proteins that move cargo along actin filaments and one of its members, myosin Va, is involved in vesicles transport. Here we studied the effect of excitotoxicity on myosin Va in cultured cerebellar granule neurons. Western blot analysis of control cultures shows a band corresponding to myosin Va as well as an 80 kDa band corresponding to its proteolytic product by calpain. When cells are exposed to glutamate (500 microM), kainate (100 microM) or NMDA (150 microM) during 3-24 h, the proteolytic processing of myosin Va is markedly increased. This proteolysis is inhibited by leupeptin (100 microM) and calpain inhibitor I (50 microM). These inhibitors also significantly improve the morphological appearance of the neurons possibly through the preservation of the cytoskeleton integrity. Our results suggest that myosin Va is a target for calpain I during an excitotoxic injury and could lead to a new area of research to address the participation of molecular motors in neurotoxicity. PMID- 15337277 TI - A report of a midwifery model for training traditional midwives in Guatemala. AB - OBJECTIVE: to describe the specific characteristics of one model of training traditional birth attendants (TBAs) in Guatemala. DESIGN: participant observation, unstructured and semi-structured interviews undertaken between 1997 and 2003 to gather the data to report on the characteristics of this training programme as it is evolving in the field. SETTING: the birth centre site of Ixmucane in Antigua, Guatemala, as well as community sites in the Departments of Saquetepequez, Chimaltenango, and Quetzaltenango in the western highlands of Guatemala. PARTICIPANTS: traditional midwives, certified nurse-midwives and certified professional midwives, as well as many allied health professionals and volunteers. INTERVENTION: training philosophy, participant selection, curriculum content, intensity, and planned follow-up are the key components of the training programme described. MEASUREMENT AND FINDINGS: 93 TBAs have received training through the development of a 150 hrs contact course for self-selected TBAs in the Midwives for Midwives Program. Formal evaluation of this training is underway but results are not yet available. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: the value of incorporating midwifery philosophy and praxis in TBA training has received scant attention in the TBA literature. This report suggests that TBA training programme characteristics are important considerations in any evaluation of training efficacy of TBAs to improve maternal-child health. PMID- 15337278 TI - Characteristics of Swedish women who do not attend childbirth and parenthood education classes during pregnancy. AB - OBJECTIVE: to investigate the attendance rate at childbirth and parenthood education classes during pregnancy in a national Swedish sample and describe the characteristics of women who did not attend. DESIGN: a cohort study utilising a postal questionnaire in early pregnancy and at 2 months after birth. SETTING: women were recruited from 97% of all antenatal clinics in Sweden at their first 'booking' visit during three different weeks spread over 1 year in 1999-2000. PARTICIPANTS: 2546 women, who were 77% of those who consented to participate in the study and 55% of all women eligible for the study. MEASUREMENT AND FINDINGS: most primiparous women (93%) attended classes and the majority of the multiparae (81%) did not. Having a native language other than Swedish was associated with non-attendance in both primiparae and multiparae (OR 2.7, 95% CI 1.3-5.4; OR 2.1, 95% CI 1.4-3.1). In addition, the following factors were associated with non attendance in the primiparae: unemployment (OR 2.0, 95% CI 1.1-3.8), smoking during pregnancy (OR 2.7, 95% CI 1.2-5.8), having considered abortion (OR 4.3, 95% CI 1.2-16.1), and having had few antenatal check-ups (OR 2.0, 95% CI 1.1 3.7). The following factors were associated with non-attendance in the multiparae: age older than 35 years (OR 1.6, 95% CI 1.1-2.3), low level of education (OR 3.6, 95% CI 2.3-5.7), and pregnancy unplanned but welcome (OR 1.5, 95% CI 1.1-2.0), having had counselling because of fear of childbirth (OR 1.6, 95% CI 1.1-2.4), and expressing a need of such counselling (OR 1.9, 95% CI 1.1 3.1). KEY CONCLUSIONS: the childbirth and parenthood education programme reached the majority of pregnant women, and that non-attendees were more disadvantaged in terms of socio-demographic background and feelings about the approaching birth. These women should be given special attention during the antenatal check-ups so that childbirth and parenthood education could be adapted to their specific needs. PMID- 15337279 TI - Multiple logistic regression models--what are they? PMID- 15337280 TI - A 'give it a go' breast-feeding culture and early cessation among low-income mothers. AB - OBJECTIVE: to examine cultural expectations and experiences of breast feeding amongst first time mothers from low-income areas, in order to improve understanding of why many cease breast feeding in the early days of their babies' lives. DESIGN: qualitative interviews were carried out with 16 women, who expressed an intention to breast feed, at 37 weeks in their pregnancy and again at 3-9 weeks postnatally. SETTING: women were interviewed in their own homes in low-income areas of North Tyneside, north-east England. FINDINGS: decisions about breast-feeding cessation were usually made within the first few days as women negotiated the pathways of informal cultures of feeding babies and the availability and quality of formal care. A 'give it a go' breast-feeding culture is identified, where women who intended to breast feed had a strong expectation of difficulties and even failure. Expertise and confidence with bottle feeding were more widespread among family and friends. The many influences on the mothers' decision-making were interconnected and contingent upon each other: if one aspect of breast feeding 'goes wrong', other reasons were often brought into play and the underlying pessimism that was felt antenatally was borne out. CONCLUSIONS: positive experiences of formal support could make a crucial difference in the early days of breast feeding. However non-breast-feeding cultures permeated and found expression in negative discourses. Support needs to take account of the cultural contexts in which mothers make decisions and the fact that breast feeding is affected by a multitude of factors simultaneously. Access to advice at the right time is a key issue for some low-income women. PMID- 15337281 TI - Breast-feeding difficulties experienced by women taking part in a qualitative interview study of postnatal depression. AB - OBJECTIVE: to explore how women experience breast-feeding difficulties. This theme emerged unexpectedly during a study of women's experiences of screening with the Edinburgh postnatal depression scale (EPDS) and subsequent care from primary health-care professionals. DESIGN: qualitative in-depth interview study. SETTING: postnatal women of 22 general practices within Oxford City Primary Care Group area. PARTICIPANTS: 39 postnatal women from a purposeful sample were interviewed at an average of 15 months postnatal. They were chosen from different general practices and with a range of emotional difficulties after birth, judged using EPDS results at eight weeks and eight months postnatal, and whether they received 'listening visits' from health visitors. MEASUREMENTS AND FINDINGS: a qualitative thematic analysis was used, including searches for anticipated and emergent themes. Fifteen women had breast-feeding difficulties. Five themes emerged which explore the difficulties. Firstly, commitment to breast feeding and high expectations of success; secondly, unexpected difficulties; thirdly, seeking professional support for difficulties; fourthly, finding a way to cope; and fifthly, guilt. KEY CONCLUSIONS: in this study breast-feeding difficulties were common, caused emotional distress and interactions with professionals could be difficult. Current breast-feeding policy, such as the 'Baby Friendly Initiative', may be a contributing factor. This needs to be explored in a further study. PMID- 15337282 TI - Conflicting ideologies as a source of emotion work in midwifery. AB - OBJECTIVE: to explore how a range of midwives experienced and managed emotion in their work. DESIGN: a qualitative study using an ethnographic approach. Data were collected in three phases using focus groups, observations and interviews. SETTING: South Wales, UK. PARTICIPANTS: Phase One: self-selected convenience sample of 27 student midwives in first and final years of 18-month (postnursing qualification) and 3-year (direct entry) programmes. Phase Two: opportunistic sample of 11 qualified midwives representing a range of clinical locations and clinical grades. Phase Three: purposive sample of 29 midwives working within one NHS Trust, representing a range of clinical locations, length of clinical experience and clinical grades. FINDINGS: community and hospital environments presented midwives with fundamentally different work settings that had diverse values and perspectives. The result was two primary occupational identities and ideologies that were in conflict. Hospital midwifery was dominated by meeting service needs, via a universalistic and medicalised approach to care; the ideology was, by necessity, 'with institution'. Community-based midwifery was more able to support an individualised, natural model of childbirth reflecting a 'with woman' ideology. This ideology was officially supported, both professionally and academically. When midwives were able to work according to the 'with woman' ideal, they experienced their work as emotionally rewarding. Conversely, when this was not possible, they experienced work as emotionally difficult and requiring regulation of emotion, i.e. 'emotion work'. KEY CONCLUSIONS: unlike findings from other studies, that have located emotion work primarily within worker/client relationships, the key source of emotion work for participants was conflicting ideologies of midwifery practice. These conflicts were particularly evident in the accounts of novice midwives (i.e. students and those who had been qualified for less than 1 year) and integrated team midwives. Both groups held a strong commitment to a 'with woman' ideology. IMPLICATIONS FOR PRACTICE: understanding the dilemmas created by conflicting occupational ideologies is important in order to improve the quality of midwives' working lives and hence the care they give to women and families. In the short term, strategies involving education and supervision may be of assistance in enabling midwives to reconcile these conflicting perspectives. However, in the long term more radical solutions may be required to address the underpinning contradictions. PMID- 15337283 TI - Midwife interactions with birth support people in Melbourne, Australia. AB - OBJECTIVE: to investigate midwife experience of the presence of birth support people in the delivery suite. PARTICIPANTS: eighteen midwives working in three hospitals in Melbourne, Australia, who were interviewed over a 3 month period in 2001. METHODS: informant views were gathered via semi-structured interviews. Interviews were transcribed and analysed for key concepts and central themes that were then compared to existing literature. FINDINGS: all midwives were supportive of the presence of birth support people to assist birthing women. The midwives were of the view that birth support people were not well prepared for the birth process, however, and did not generally have clear ideas about what support was necessary or required. Midwives indicated that the presence of birth support people in the birth room added to their workload and sometimes increased the difficulty of providing care to the birthing woman. KEY CONCLUSIONS: while the presence of birth support people is now commonplace in most Western medical systems, the presence of these birth support people may result in additional workload and stress for midwives. This aspect of the work of midwives requires greater investigation. PMID- 15337284 TI - The impact of pregnancy weight and glucose on the metabolic health of mother and child in the south west of the UK. AB - OBJECTIVE: to explore relationships between maternal pre-pregnancy weight, third trimester glucose, baby birth weight, weight and metabolic health of the mother and child 5 years after birth. DESIGN: an observational study set within a non intervention, longitudinal cohort study looking at insulin resistance in children. SETTING: a teaching hospital in the south west of the United Kingdom. PARTICIPANTS: 300 mothers and their five-year-old children from randomly selected Plymouth schools, stratified according to socioeconomic status. MEASUREMENTS: were obtained from obstetric records maternal pre-pregnant weight, random and fasting third trimester blood glucose, baby birth weight. Five years later the following measurements were made of the mother and child: height, weight, glucose and insulin resistance. FINDINGS: five years after the pregnancy, 33% of the mothers were overweight, with an additional 19% obese. In the children 13% of boys were overweight (4% obese), and in the girls, 26% were overweight (5% obese). In the five-year-old children, weight (r=0.28, p<0.001) but not birth weight (r=0.03, p=0.573), correlated with insulin resistance. Maternal pre pregnant weight was related to both random and fasting third trimester glucose, and to insulin resistance 5 years later. Third trimester fasting glucose, even within a normal range, was a better predictor than random glucose of the baby's birth weight (r=0.39, p=0.044) and the mother's future insulin resistance (r=0.67, p<0.001). No maternal measures predicted insulin resistance in the child at 5 years. CONCLUSIONS: maternal weight had an important influence on the gestational environment, and predicted insulin resistance 5 years later. Fasting glucose, even within the reference range, was a better predictor than random glucose of the baby's birth weight and the mother's future insulin resistance. IMPLICATIONS FOR PRACTICE: these concern the importance of pre-conception weight management, and support replacement of routine random glucose sampling during the third trimester with an earlier, fasting measurement. PMID- 15337285 TI - The International Confederation of Midwives Americas Regional Conference Port of Spain, Trinidad, April 2004. PMID- 15337286 TI - Arylsulfonyl acridinyl derivatives acting on Plasmodium falciparum. AB - Several arylacridinyl sulfones have been synthesized and their antimalarial action was tested on Plasmodium falciparum. PABA (para-aminobenzoic acid) has no antagonistic effect with these compounds as opposed to the observed effect with dapsone and sulfonamides previously studied. A possible relationship between the ability of cleavage of the S-9C acridinic bond and activity is suggested. PMID- 15337287 TI - 3D QSAR studies on antimalarial alkoxylated and hydroxylated chalcones by CoMFA and CoMSIA. AB - The 3D QSAR analyses of antimalarial alkoxylated and hydroxylated chalcones were first conducted by Comparative molecular field analysis (CoMFA) and Comparative similarity indices analysis (CoMSIA) to determine the factors required for the activity of these compounds. Satisfactory results were obtained after performing a leave-one-out (LOO) cross-validation study with cross-validation q(2) and conventional r(2) values of 0.740 and 0.972 by the CoMFA model, 0.714 and 0.976 by the CoMSIA model, respectively. The results provided the tools for predicting the affinity of related compounds, and for guiding the design and synthesis of novel and more potent antimalarial agents. PMID- 15337288 TI - Synthesis and cytotoxicity study of alkannin derivatives. AB - Alkannin derivatives (3-19) were prepared through the reaction of beta,beta dimethylacrylalkannin (1), the most abundant isohexenylnaphthazarin isolated from the roots of Arnebia euchroma, with different types of nucleophiles such as amines and thiols in the absence or presence of a reducing agent. The cytotoxicities of 1-8, 10-14 and 19 against four human carcinoma cell line (GLC 82, CNE2, Bel-7402, K-562) were found to be markedly higher than that of the naturally occurring beta,beta-dimethylacrylalkannin (1) and acetylalkannin (2). This study also shed light on the understanding of the biological activities in terms of the chemical reactivity of alkannins. PMID- 15337289 TI - Cytotoxic activity of new neodymium (III) complexes of bis-coumarins. AB - Complexes of neodymium (III) with bis-coumarins: 3,3'-benzylidene-bis(4-hydroxy 2H-1-benzopyran-2-one); bis(4-hydroxy-2-oxo-2H-chromen-3-yl)-piridin-2-yl methane; bis(4-hydroxy-2-oxo-2H-chromen-3-yl)-piridin-4-yl-methane; bis(4-hydroxy 2-oxo-2H-chromen-3-yl)-(1H-pyrazol-3-yl)-methane were synthesized by reaction of neodymium (III) salt and the ligands, in amounts equal to metal:ligand molar ratio of 1:2. The complexes were prepared by adding an aqueous solution of neodymium (III) salt to an aqueous solution of the ligand subsequently raising the pH of the mixture gradually to ca. 5.0 by adding dilute solution of sodium hydroxide. The neodymium (III) complexes with bis-coumarins were characterized by different physicochemical methods-elemental analysis, IR-, (1)H- and (13)C-NMR spectroscopies and mass-spectral data. The spectral data of neodymium (III) complexes were interpreted on the basis of comparison with the spectra of the free ligands. This analysis showed that in the Nd (III) complexes the ligands coordinated to the metal ion through both deprotonated hydroxyl groups. On the basis of the nu(C=O) red shift observed, participation of the carbonyl groups in the coordination to the metal ion was also suggested. Cytotoxic screening by MTT assay was carried out. The complexes were tested on HL-60, HL-60/Dox and SKW-3 cell lines. The overall results from the preliminary screening program revealed that all of the new Nd (III) complexes reach 50% inhibition of the malignant cells proliferation and thus could be considered as biologically active. On the basis of the IC(50) values obtained compounds Nd(L(1))(OH).H(2)O and Nd(L(3))(OH).2H(2)O were found to exert superior activity in comparison to the remaining complexes. PMID- 15337290 TI - One pot synthesis of thiazolodihydropyrimidinones and evaluation of their anticancer activity. AB - 2-(5-Arylfurfurylidene/5-nitrofurfurylidene)-5-aryl-7-(2,4-dichloro-5 fluorophenyl)-5H-thiazolo[2,3-b]-pyrimidin-2(1H)-ones 5 and 6 are synthesized by a novel three component reaction of 4,6-diarylpyrimidino-2(1H)-thiones 4, monochloroacetic acid, arylfurfuraldehydes and 5-nitro-2-furfuraldenediacetate, respectively. The newly synthesized compounds are characterized by elemental analysis, IR, (1)H NMR and mass spectral studies. These compounds exhibited in vitro antitumour activity with moderate to excellent growth inhibition against a panel of 60 cell lines of leukemia, non-small cell lung cancer melanoma, ovarian cancer, prostate cancer and breast cancer. PMID- 15337291 TI - N-Benzyl-2-chloroindole-3-carboxylic acids as potential anti-inflammatory agents. Synthesis and screening for the effects on human neutrophil functions and on COX1/COX2 activity. AB - The synthesis of N-benzyl-2-chloroindole-3-carboxylic acids related to indomethacin is reported. These compounds were tested on in vitro human neutrophil activation. Some of them, more soluble in water, were tested to define the influence on prostaglandin biosynthesis via inhibition of cyclooxygenases (COX1 and COX2) by a chemiluminescent method suitable for fast screening. Several derivatives showed inhibitory effects and in some cases were more active than the parent compound. PMID- 15337292 TI - Synthesis and antimicrobial activities of some new 1-(5-phenylamino [1,3,4]thiadiazol-2-yl)methyl-5-oxo-[1,2,4]triazole and 1-(4-phenyl-5-thioxo [1,2,4]triazol-3-yl)methyl-5-oxo- [1,2,4]triazole derivatives. AB - Acetic acid ethyl esters containing 5-oxo-[1,2,4]triazole ring (2) were synthesized by the condensation of compounds 1a-f with ethyl bromoacetate in basic media. The reaction of compounds 2a-f with hydrazine hydrate led to the formation of acid hydrazides (3a-f). The treatment of compounds 3 with two divers aromatic aldehydes resulted in the formation of arylidene hydrazides as cis-trans conformers (4a,c,e,f, 5a,e,f). The thiosemicarbazide derivatives (6a,c,d,f) were afforded by the reaction of corresponding compounds 3 with phenylisothiocyanate. The treatment of compounds 6a,c,d,f with sulfuric acidic caused the conversion of side-chain of compounds 6a,c,d,f into 1,3,4-thiadiazol ring; thus, compounds 7a,c,d,f were obtained. On the other hand, the cyclization of compounds 6a,c,d,f in the presence of 2 N NaOH resulted in the formation of compounds 8a,c,d,f containing two [1,2,4]triazole rings which are linked to each other via a methylene bridge. Compounds 4a, f, 5a, 7a, d, f, 8a and d have shown antimicrobial activity against one or more microorganism, but no antifungal activity has been observed against yeast like fungi. Also inhibitory effect on mycelial growth by compounds 4e, 7d and 8f has been observed. Compounds 4c and 5f were found to possess antitumor active towards breast cancer. PMID- 15337293 TI - Ring-substituted imidazoles as a new class of anti-tuberculosis agents. AB - We describe in vitro anti-Mycobacterium tuberculosis activities of ring substituted-1H-imidazole-4-carboxylic acid derivatives (1-6), and 3-(2-alkyl-1H imidazol-4-yl)-propionic acid derivatives (7-13) against drug-sensitive and drug resistant M. tuberculosis H37Rv strains. The most effective analogues, 2f (R=R(1)=c-C(5)H(9)), and 2h (R=R(1)=c-C(6)H(11)) have produced >90% inhibition at a concentration of <6.25 microg/ml in the drug-sensitive screen. Upon further evaluation against drug-resistant strains, both analogues 2f and 2h produced an MIC value of 25.0 microg/ml. The observation of significant anti-tuberculosis activity in some of these analogues describes the discovery of novel ring substituted-1H-imidazole-4-carboxylic acid ethyl esters as a new class of anti tuberculosis agents. PMID- 15337294 TI - Oxidative stress and inhibition of oxidative phosphorylation induced by peroxynitrite and nitrite in rat brain subcellular fractions. AB - Nitrite and nitrate, two endogenous oxides of nitrogen, are toxic in vivo. Furthermore, the reaction of superoxide (produced by all aerobic cells) with nitric oxide (NO) generates peroxynitrite, a potent oxidizing agent, that can cause biological oxidative stress. Using subcellular fractions from rat brain hemispheres we studied oxidative stress induced by these nitrogen compounds with special emphasis on nitrite. The consumption of Vitamin C (ascorbate) and Vitamin E (alpha tocopherol), two of the important nutritional antioxidants, was followed in synaptosomes (nerve-ending particles) and mitochondria along with changes in parameters of mitochondrial oxidative phosphorylation. Nitrite, but not nitrate, oxidized ascorbate without oxidizing alpha tocopherol in both synaptosomes and mitochondria whereas peroxynitrite oxidized both ascorbate and alpha tocopherol. Functionally, both nitrite and peroxynitrite inhibited mitochondrial oxidative phosphorylation. Nitrite was less potent than peroxynitrite when the effects of equal concentrations of the two were compared. However, since nitrite is much more stable than peroxynitrite the impact of nitrite as an oxidant in vivo could be as much or even more significant than peroxynitrite. Nitrate would not have similar action unless it is reduced to nitrite. It is possible that nitrite may impair oxidative phosphorylation through modulating levels of nitric oxide, changing the activity of heme proteins or a mild uncoupling of mitochondria. PMID- 15337295 TI - Ethanol acutely decreases astroglial gap junction permeability in primary cultures from defined brain regions. AB - The acute effect of hyperosmotic ethanol on gap junction permeability was examined in astroglial cells in primary culture from five different brain regions. Gap junction permeability was analyzed by measuring dye spreading from cell to cell with the low molecular weight dye Lucifer Yellow. Ethanol concentrations 25-300 mM significantly decreased dye spreading in cultures from the cerebral cortex in a dose-dependent but time-independent manner for up to 60 min. Besides cerebral cortex, exposure to 150 mM ethanol decreased dye spreading in astroglial cultures from the hippocampus and from the brain stem, while cultures from the olfactory bulb and from the hypothalamus were not significantly affected. The ethanol-induced decrease in dye spreading in cultures from the cerebral cortex was not mediated through changes in cell volume, osmolarity, protein kinase C (PKC) phosphorylation, intracellular pH, or intracellular calcium concentration ([Ca(2+)](i)). The decrease in dye spreading was abolished upon incubation in sodium-reduced buffer, and after blockage of the Na(+)/K(+)/2Cl(-) cotransporter with furosemide. The results presented here indicate that ethanol-mediated decrease in dye spreading is directly or indirectly dependent on sodium. PMID- 15337296 TI - Central and peripheral changes in catecholamine biosynthesis and turnover in rats after a short period of ozone exposure. AB - We investigated in rat the effects of ozone exposure (0.7 ppm) for 5 h on the catecholamine biosynthesis and turnover in sympathetic efferents and various brain areas. For this purpose, the activity of tyrosine hydroxylase, the rate limiting enzyme in catecholamine biosynthesis, was assessed in superior cervical ganglia and in two major noradrenergic cell groups, A2 and A6 (locus coeruleus). Tyrosine hydroxylase activity was estimated in vivo by measuring the accumulation of l-dihydroxyphenylalanine after pharmacological blockade of L-aromatic acid decarboxylases by NSD-1015 (100 mg/kg i.p.). The catecholamine turnover rate was measured after inhibition of tyrosine hydroxylase by alpha-methyl-para-tyrosine (AMPT, 250 mg/kg, i.p., 2.5 h) in peripheral sympathetic target organ (heart and lungs) as well as in some brain catecholamine terminal areas (cerebral cortex, hypothalamus and striatum). Ozone caused differential effects according to the structure. Catecholamine biosynthesis was stimulated in superior cervical ganglia (+44%, P < 0.05) and caudal A2 subset (+126%, P < 0.01), whereas catecholamine turnover was increased in heart (+183%, P < 0.01) and cortex (+22%, P < 0.05). On the other hand, catecholamine turnover was inhibited in lungs (-53%, P < 0.05) and striatum (-24%, P < 0.05). A brief exposure to ozone, at a concentration chosen to mimic pollution level encountered in urban areas, can modulate catecholamine biosynthesis and utilization rate in the sympathetic and central neurones. PMID- 15337297 TI - Botulinum toxin A inhibits ATP release from bladder urothelium after chronic spinal cord injury. AB - The effects of mechanoreceptor stimulation and subsequent ATP release in spinal cord injured and normal bladders was examined to demonstrate if spinal cord injury (SCI) modulates the basal or evoked release of ATP from bladder urothelium and whether intravesical botulinum toxin A (BTX-A) administration inhibits urothelial ATP release, a measure of sensory nerve activation. A Ussing chamber was used to isolate and separately measure resting and mechanoreceptor evoked (e.g. hypoosmotic stimulation) ATP release from urothelial and serosal sides of the bladder. Following spinal cord injury, resting urothelial release of ATP was ninefold higher than that of normal rats. Botulinum toxin A instillation did not significantly affect the resting release of ATP after spinal cord injury. Evoked ATP release following hypoosmotic stimulation was significantly higher in chronic spinal cord injured compared to normal rat bladders. However, botulinum toxin A treatment markedly reduced ATP release in spinal cord injured bladders by 53% suggesting that ATP release by mechanoreceptor stimulation, as opposed to basal release, occurs by exocytotic mechanisms. In contrast, there was no significant difference in basal or evoked ATP release from bladder serosa following spinal cord injury. Moreover, intravesical instillation of botulinum toxin A did not affect ATP release from the serosal side after spinal cord injury, suggesting that its effects were confined to the urothelial side of the bladder preparation. In summary: (1) increased release of ATP from the urothelium of spinal cord injured bladders may contribute to the development of bladder hyperactivity and, (2) mechanoreceptor stimulated vesicular ATP release, as opposed to basal non vesicular release of ATP, is significantly inhibited in spinal cord injured bladders by intravesical instillation of botulinum toxin A. These results may have important relevance in our understanding of the mechanisms underlying plasticity of bladder afferent pathways following SCI. PMID- 15337298 TI - Partial bilateral mesencephalic lesions affect D1 but not D2 binding in both the striatum and cortex. AB - The substantia nigra pars compacta (SNc) and the ventral tegmental area (VTA) are the two major mesencephalic dopaminergic systems. Mesencephalic dopamine denervation is followed by long-term modifications in striatum and cortex that preserve dopamine functions. Here, we have studied the impact of isolated bilateral 6-hydroxydopamine lesioning of the SNc or the VTA on D(1) and D(2) dopamine receptor binding in striatal and cortical areas of rat. Neither SNc nor VTA bilateral partial lesioning changed D(2) binding at the striatal or cortical level. Intriguingly, only VTA lesioning increased D(1) binding in the cortex, whereas both bilateral partial lesioning of the SNc or the VTA increased striatal D(1) binding. This suggests that increased cortical D(1) binding could be an indicator of VTA lesioning. Further behavioural experiments may explain the pathophysiological meaning of increased cortical D(1) binding, and determine whether this observation is involved in compensatory mechanisms. PMID- 15337299 TI - Expression of vanilloid VR1 receptor in PC12 cells. AB - Capsaicin, a pungent ingredient of chili pepper, activates vanilloid receptor subtype 1 (VR1), which is a nonselective cation channel with high Ca(2+) permeability. Although VR1 and its splice variant are highly expressed in sensory neurons, they are expressed in neuronal cells in brain and peripheral non neuronal cells. In this study, we investigated whether VR1 is expressed in PC12 cells, rat pheochromocytoma. Capsaicin at concentrations above 100 microM induced an increase in intracellular free Ca(2+) concentrations by influx from extracellular spaces, and the effect was blocked by capsazepine, a selective antagonist of VR1. VR1 transcript and protein were detected by reverse transcription-polymerase chain reaction and Western blotting analysis, respectively. Immunocytochemical analysis revealed that VR1 protein was expressed in the cytosol and the plasma membrane of PC12 cells, and treatment with the antisense oligonucleotide for VR1 decreased the expression. VR1 in PC12 cells showed different characters from that in sensory neurons; capsaicin concentration dependency and heat- and nerve growth factor-sensitivities. These results suggested that VR1 was functionally expressed in PC12 cells. The usefulness of PC12 cell line for studying functions and/or expression of VR1 is discussed. PMID- 15337300 TI - GABAA receptor beta subunit mRNA expression in the human alcoholic brain. AB - A competitive RT-PCR assay was used to quantify the expression of the GABA(A) receptor beta(1), beta(2) and beta(3) isoform mRNA transcripts in the superior frontal cortex and motor cortex of 21 control and 22 alcoholic cases. A single set of primers was designed that permitted amplification of all three transcripts and the internal standard simultaneously; differentiation of the individual transcripts was achieved by restriction enzyme digestion. Construction of a standard curve, using the internal standard and a concentration range of beta(2) cRNA-enabled quantitation of mRNA expression levels. No significant difference in mRNA expression was found between the control and alcoholic case groups in either the superior frontal or motor cortex for the beta(2) or beta(3) isoforms. A significant interaction was found between isoform and area, although, the two case groups did not partition on this measure. The interaction was due to a significant difference between superior frontal and motor cortex for the beta(3) isoform; this regional comparison was not significant for beta(2) mRNA. Age at death and post-mortem delay (PMD) had no significant effect on beta mRNA expression in either case group in either region. A beta(1) signal could not be detected in the RT-PCR assay. PMID- 15337301 TI - Tricarboxylic acid cycle enzymes following thiamine deficiency. AB - Thiamine (Vitamin B1) deficiency (TD) leads to memory deficits and neurological disease in animals and humans. The thiamine-dependent enzymes of the tricarboxylic acid (TCA) cycle are reduced following TD and in the brains of patients that died from multiple neurodegenerative diseases. Whether reductions in thiamine or thiamine-dependent enzymes leads to changes in all TCA cycle enzymes has never been tested. In the current studies, the pyruvate dehydrogenase complex (PDHC) and all of enzymes of the TCA cycle were measured in the brains of TD mice. Non-thiamine-dependent enzymes such as succinate dehydrogenase (SDH), succinate thiokinase (STH) and malate dehydrogenase (MDH) were altered as much or more than thiamine-dependent enzymes such as the alpha-ketoglutarate dehydrogenase complex (KGDHC) (-21.5%) and PDHC (-10.5%). Succinate dehydrogenase (SDH) activity decreased by 27% and succinate thiokinase (STH) decreased by 24%. The reductions in these other enzymes may result from oxidative stress because of TD or because these other enzymes of the TCA cycle are part of a metabolon that respond as a group of enzymes. The results suggest that other TCA cycle enzymes should be measured in brains from patients that died from neurological disease in which thiamine-dependent enzymes are known to be reduced. The diminished activities of multiple TCA cycle enzymes may be important in our understanding of how metabolic lesions alter brain function in neurodegenerative disorders. PMID- 15337302 TI - Changes in extracellular dopamine in the rat globus pallidus induced by typical and atypical antipsychotic drugs. AB - Typical antipsychotic drugs with a high extrapyramidal motor side-effects liability markedly increase extracellular dopamine in the caudate-putamen, while atypical antipsychotic drugs with a low incidence of extrapyramidal motor side effects have less pronounced stimulating actions on striatal dopamine. Therefore, it has been suggested that the extrapyramidal motor side-effects liability of antipsychotic drugs (APD) is correlated with their ability to increase extracellular dopamine in the caudate-putamen. The globus pallidus (GP) is another basal ganglia structure probably mediating extrapyramidal motor side effects of typical antipsychotic drugs. Therefore, the present study sought to determine whether extracellular dopamine in the globus pallidus might be a further indicator to differentiate neurochemical actions of typical and atypical antipsychotic drugs. Using in vivo microdialysis we compared effects on pallidal dopamine induced by typical and atypical antipsychotic drugs in rats. Experiment I demonstrated that systemic administration of haloperidol (1 mg/kg; i.p.) and clozapine (20 mg/kg; i.p.) induced a significant pallidal dopamine release to about 160 and 180% of baseline, respectively. Experiment II revealed that reverse microdialysis of raclopride and clozapine using a cumulative dosing regimen did not stimulate extracellular dopamine in the globus pallidus if low (1microM) or intermediate (10 and 100 microM) concentrations were used. Only at a high concentration (1,000 microM), raclopride and clozapine induced a significant pallidal dopamine release to about 130 and 300% of baseline values, respectively. Thus, effects of typical and atypical antipsychotic drugs on pallidal dopamine were similar and thus, may not be related to their differential extrapyramidal motor side-effects liability. Furthermore, the finding that reverse microdialysis of raclopride over a wide range of concentrations did not stimulate pallidal dopamine concentrations tentatively suggests that pallidal dopamine release under basal conditions is not regulated by D2 autoreceptors. PMID- 15337303 TI - Heterogeneous competition of Kv1 channel toxins with kaliotoxin for binding in rat brain: autoradiographic analysis. AB - The alpha-subunits of Kv1 channels display characteristic distributions and restricted co-assembly in mammalian brain. The heterogeneous composition of Kv1 channels has made it difficult to use specific toxins to label brain structures. We used autoradiography to analyse the competitive behaviour of three Kv1 channel toxins--alpha-dendrotoxin, kaliotoxin, and mast cell degranulating peptide--for binding to kaliotoxin binding sites in various brain structures. IC(50) varied considerably between brain regions (by up to three orders of magnitude) for each ligand. alpha-dendrotoxin and kaliotoxin competed equally in some regions and to different extents in others, identifying two types of structure. Mast cell degranulating peptide competed with (125)I-kaliotoxin less efficiently than alpha dendrotoxin and kaliotoxin, in all regions. Thus, differences in the capacity of these three toxins to bind to kaliotoxin binding sites provide evidence of major differences in the composition of the Kv1 channels constituting the kaliotoxin binding sites. PMID- 15337304 TI - Modulation of dopamine release by the nicotinic agonist epibatidine in the frontal cortex and the nucleus accumbens of naive and chronic nicotine treated rats. AB - The effect of the nicotinic acetylcholine receptors (nAChRs) agonist (+/ )epibatidine on the modulation of dopamine (DA) release was investigated by microdialysis in vivo in the frontal cortex and the nucleus accumbens of naive and chronic nicotine-treated awake rats. (+/-)Epibatidine (2.5 microg/kg, s.c.), contrary to (-)nicotine (0.5 mg/kg, s.c.), decreased the extracellular concentrations of DA in the brain of naive rats. Subchronic nicotine treatment (0.45 mg/kg, s.c., twice daily for 7 days) attenuated the (+/-)epibatidine induced decrease in the DA level. The extracellular concentrations of the DA metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were elevated by (+/-)epibatidine administration in both naive and subchronic treated rats. The findings suggest that the decrease in DA extracellular concentrations induced by the high affinity nAChRs agonist (+/-)epibatidine might be due to inactivation of nAChRs, which can be overcome by subchronic treatment with nicotine. Different mechanisms in modulation of DA release appears to be involved in the rat brain by (+/-)epibatidine compare to (-)nicotine. PMID- 15337305 TI - GABA receptor modulation of 5-HT neuronal firing: characterization and effect of moderate in vivo variations in glucocorticoid levels. AB - Evidence from electrophysiological studies suggests that 5-HT neuronal firing in the dorsal raphe nucleus (DRN) may be regulated by both GABA(A) and GABA(B) receptors. Here, we addressed the question of whether the activity of individual 5-HT neurons is regulated by both GABA(A) and GABA(B) receptors. In addition, we examined the concentration-response relationships of GABA(A) and GABA(B) receptor activation and determined if GABA receptor regulation of 5-HT neuronal firing is altered by moderate alterations in circulating corticosterone. The activity of 5 HT neurons in the DRN of the rat was examined using in vitro extracellular electrophysiology. The firing of all individual neurons tested was inhibited by both the GABA(A) receptor agonist 4,5,6,7-tetrahydroisoxazolo-[5,4-c]-pyridin-3 ol hydrochloride (THIP) (25 microM) and the GABA(B) receptor agonist baclofen (1 microM). Responses to THIP (5, 10, 25 microM) and baclofen (1, 3, 10 microM) were concentration dependent and attenuated by the GABA(A) and GABA(B) receptor antagonists, bicuculline (50 microM) and phaclofen (200 microM), respectively. To examine the effects of corticosterone on the sensitivity of 5-HT neurons to GABA receptor activation, experiments were conducted on adrenalectomized animals with corticosterone maintained for two weeks at either a low or moderate level within the normal diurnal range. These changes in corticosterone levels had no significant effects on the 5-HT neuronal response to either GABA(A) or GABA(B) receptor activation. The data indicate that the control of 5-HT neuronal activity by GABA is mediated by both GABA(A) and GABA(B) receptors and that this control is insensitive to moderate changes in circulating glucocorticoid levels. PMID- 15337306 TI - Presynaptic dopaminergic properties of differentiated mouse embryonic stem cells. AB - This study characterized the presynaptic dopaminergic properties of neuronally differentiated mouse embryonic stem (ES) cells. Approximately 30% of the ES cells expressed tyrosine hydroxylase (TH) immunoreactivity when co-cultured with PA6 cells. These cultures expressed high affinity, sodium-dependent dopamine uptake as well as depolarization-induced and calcium-dependent dopamine release of this transmitter. These and other important dopaminergic genes found expressed in these cultures by RT-PCR included Nurr1, vesicular monoamine transporter 2 (VMAT2), TH, dopamine transporter (DAT), and glial cell line-derived neurotrophic factor (GDNF) receptors c-Ret and GFRalpha1. These results demonstrate that differentiated ES cells have the presynaptic functions for maintaining dopaminergic homeostasis, which may be essential for their long-term use in restoring CNS levels of this transmitter. PMID- 15337307 TI - Glutaric acid stimulates glutamate binding and astrocytic uptake and inhibits vesicular glutamate uptake in forebrain from young rats. AB - Glutaric acidemia type I (GA I) is an inherited neurometabolic disorder caused by glutaryl-CoA dehydrogenase deficiency, which leads to accumulation in body fluids and in brain of predominantly glutaric acid (GA), and to a lesser extent of 3 hydroxyglutaric and glutaconic acids. Neurological presentation is common in patients with GA I. Although the mechanisms underlying brain damage in this disorder are not yet well established, there is growing evidence that excitotoxicity may play a central role in the neuropathogenesis of this disease. In the present study, preparations of synaptosomes, synaptic plasma membranes and synaptic vesicles, as well as cultured astrocytes from rat forebrain were exposed to various concentrations of GA for the determination of the basal and potassium induced release of [(3)H]glutamate by synaptosomes, Na(+)-independent glutamate binding to synaptic membranes and vesicular glutamate uptake and Na(+)-dependent glutamate uptake into astrocytes, respectively. GA (1-100 nM) significantly stimulated [(3)H]glutamate binding to brain plasma membranes (40-70%) in the absence of extracellular Na(+) concentrations, reflecting glutamate binding to receptors. Furthermore, this stimulatory effect was totally abolished by the metabotropic glutamate ligands DHPG, DCG-IV and l-AP4, attenuated by the ionotropic non-NMDA glutamate receptor agonist AMPA and had no interference of the NMDA receptor antagonist MK-801. Moreover, [(3)H]glutamate uptake into synaptic vesicles was inhibited by approximately 50% by 10 and 100 nM GA and Na(+)-dependent [(3)H]glutamate uptake by astrocytes was significantly increased (up to 50%) in a dose-dependent manner (maximal stimulation at 100 microM GA). In contrast, synaptosomal glutamate release was not affected by the acid at concentrations as high as 1 mM. These results indicate that the inhibition of glutamate uptake into synaptic vesicles by low concentrations GA may result in elevated concentrations of the excitatory neurotransmitter in the cytosol and the stimulatory effect of this organic acid on glutamate binding may potentially cause excitotoxicity to neural cells. Finally, taken together these results and previous findings showing that GA markedly decreases synaptosomal glutamate uptake, it is possible that the stimulatory effect of GA on astrocyte glutamate uptake might indicate that astrocytes may protect neurons from excitotoxic damage caused by GA by increasing glutamate uptake and therefore reducing the concentration of this excitatory neurotransmitter in the synaptic cleft. PMID- 15337308 TI - Evidence that 3-hydroxyglutaric acid interacts with NMDA receptors in synaptic plasma membranes from cerebral cortex of young rats. AB - Neurological symptoms are common in patients with glutaric acidemia type I (GA I). Although the pathophysiology of this disorder is not yet fully established, 3 hydroxyglutaric acid (3-HGA), which accumulates in affected patients, has recently been demonstrated to be excitotoxic to embryonic chick and neonatal rat neurons probably via NMDA glutamate receptors. In the present study, we investigated the in vitro effects of 3-HGA on the [(3)H]glutamate and [(3)H]MK 801 (dizocilpine) binding to rat synaptic plasma membranes from cerebral cortex of young rats in order to elucidate the interactions of 3-HGA with glutamate receptors and its possible contribution to the in vitro excitotoxic properties of 3-HGA. 3-HGA (10-100 microM) significantly decreased Na(+)-dependent (up to 62%) and Na(+)-independent (up to 30%) [(3)H]glutamate binding to synaptic membranes, reflecting a possible competition between glutamate and 3-HGA for the glutamate transporter and receptor sites, respectively. Since a decrease in Na(+) independent glutamate binding might represent an interaction of 3-HGA with glutamate receptors, we next investigated whether 3-HGA interacts with NMDA receptors by adding NMDA alone or combined with 3-HGA and measuring Na(+) independent [(3)H]glutamate binding to synaptic membranes (binding to receptors). We verified that 3-HGA and NMDA, at 10 and 100 microM concentrations, decreased glutamate binding by up to 20 and 45%, respectively, and that the simultaneous addition of both substances did not provoke an additive effect, implying that they bind to NMDA receptors at the same site. Furthermore, the binding of the NMDA-channel blocker [(3)H ]MK-801 was significantly increased (approximately 32 40%) by 10 and 100 microM 3-HGA, implying that 3-HGA was able to open the NMDA channel allowing MK-801 binding, which is a characteristic of NMDA agonists. On the other hand, glutamate had a much higher stimulatory effect on this binding (180% increase), reflecting its strong NMDA agonist property. Furthermore, the simultaneous addition of 3-HGA and glutamate provoked an additive stimulatory effect on [(3)H]MK-801 binding to the NMDA receptor. These data indicate that, relatively to glutamate, 3-HGA is a weak agonist of NMDA receptors. Finally, we demonstrated that 3-HGA provoked a significant increase of extracellular calcium uptake by cerebral cortex slices, strengthening therefore, the view that 3-HGA activates NMDA receptors. The present study therefore, demonstrates at the molecular level that 3-HGA modulates glutamatergic neurotransmission and may explain previous findings relating the neurotoxic actions of this organic acid with excitotoxicity. PMID- 15337309 TI - A new GLT1 splice variant: cloning and immunolocalization of GLT1c in the mammalian retina and brain. AB - We have identified a novel carboxyl-terminal splice-variant of the glutamate transporter GLT1, which we denote as GLT1c. Within the rat brain only low levels of protein and message were detected, protein expression being restricted to end feet of astrocytes apposed to blood vessels or some astrocytes adjacent to the ventricles. Conversely, within the retina, this variant was selectively and heavily expressed in the synaptic terminals of both rod- and cone-photoreceptors in both humans and rats. Double-immunolabelling with antibodies to the carboxyl region of GLT1b/GLT1v, which is strongly expressed in apical dendrites of bipolar cells and in cone photoreceptors revealed that in the rat GLT1c was co-localised with GLT1b/GLT1v in cone photoreceptors but not with GLT1b/GLT1v in bipolar cells. GLT1c expression was developmentally regulated, only appearing at around postnatal day 7 in the rat retina, when photoreceptors first exhibit a dark current. Since the glutamate transporter EAAT5 is also expressed in terminals of rod photoreceptor terminals these data indicate that rod photoreceptors express two glutamate transporters with distinct properties. Similarly, cone photoreceptors express two glutamate transporters. We suggest that differential usage of these transporters by rod and cone photoreceptors may influence the kinetics of glutamate transmission by these neurons. PMID- 15337310 TI - Degradation of survival motor neuron (SMN) protein is mediated via the ubiquitin/proteasome pathway. AB - Homozygous deletion or mutation in the survival motor neuron (SMN)1 gene causes proximal spinal muscular atrophy (SMA), whereas SMN2 acts as a modifying gene that can influence the severity of SMA. It has been suggested that restoration of the SMN protein level in neuronal cells may prevent cell loss and may be helpful for treatment of SMA. Recent studies indicate that the ubiquitin/proteasome pathway is a major system for proteolysis of intracellular proteins. In this study, we investigate whether SMN protein is degraded via the ubiquitin/proteasome pathway. Primary fibroblasts were established from the skin biopsies of SMA patients and the effect of a proteasome inhibitor MG132 and lysosome inhibitor NH(4)Cl on SMN protein level was examined. We found that MG132, but not NH(4)Cl, significantly increased the amount and nuclear accumulation of SMN protein in SMA patient's fibroblasts. Immunoprecipitation/western blot analysis indicated that SMN protein was ubiquitinated in cells. In vitro protein ubiquitination assay also demonstrated that SMN protein could be conjugated with ubiquitin. Taken together, we have provided clear evidences that degradation of SMN protein is mediated via the ubiquitin/proteasome pathway and suggest that proteasome inhibitors may up regulate SMN protein level and may be useful for the treatment of SMA. PMID- 15337311 TI - Astrocytes repress the neuronal expression of GLAST and GLT glutamate transporters in cultured hippocampal neurons from embryonic rats. AB - Glutamate extracellular levels are regulated by specific transporters. Five subtypes have been identified. The two major ones, GLAST and GLT (glutamate transporters 1 and 2, respectively), are localized in astroglia in normal mature brain. However, in neuron-enriched hippocampal cultures, these proteins are expressed in neurons during the early in vitro development (Plachez et al., 2000). Here, we show that, in these cultures, GLAST and GLT neuronal expression is transient and no longer observed after 7 days in vitro, a stage at which the few astrocytes present in the culture are maturing. Moreover, we demonstrate that these few astrocytes are responsible for the repression of this neuronal expression. Indeed, addition of conditioned medium prepared from primary cultures of hippocampal astrocytes, to cultured hippocampal neurons, rapidly leads to the suppression of neuronal GLAST expression, without affecting neuronal GLT expression. However, when neurons are seeded and co-cultured on a layer of hippocampal astrocytes, they do not develop any immunoreactivity towards GLAST or GLT antibodies. Altogether, these results indicate that glia modulate the expression of GLAST and GLT glutamate transporters in neurons, via at least two distinct mechanisms. Neuronal GLAST expression is likely repressed via the release or the uptake of soluble factors by glia. The repression of neuronal GLT expression probably results from glia-neuron interactions. This further reinforces the fundamental role of direct or indirect neuron-glia interactions in the development of the central nervous system. PMID- 15337312 TI - Deficiency of the Mre11 DNA repair complex in Alzheimer's disease brains. AB - The hallmarks of Alzheimer's disease (AD) brains are deposition of Abeta plaques, appearance of neurofibrillary tangles, and extensive loss of neuronal cells. While Abeta plaques and neurofibrillary tangles play a significant role in the pathogenic dysfunction of neurons, factors that accelerate the neurodegenerating process remain to be defined. One set of factors examined in this study is the Mre11 protein complex, composed of Rad50, Mre11 and Nbs1. This protein complex plays an essential role in cellular responses to DNA damage, such as initiating cell cycle checkpoints and repairing damaged DNA. A defect in any component of this protein complex is detrimental to cells. Recently, several groups have observed abnormal cell cycle regulation and/or accumulated DNA damage in AD neurons. These pathological alterations could conceivably be caused or exacerbated by any compromise in the Mre11 complex. In this study, we compared the levels of the Mre11 complex proteins in brain samples from AD and age-matched non-dementia controls. We show for the first time that the Mre11 complex proteins are present in neurons of the adult human cortex and cerebellum. These proteins were found substantially reduced in the neurons of AD cortex. Our finding suggests that the loss of the Mre11 complex may be associated with the pathogenesis of AD. PMID- 15337313 TI - Correlation between synaptogenesis and the PTEN phosphatase expression in dendrites during postnatal brain development. AB - The PTEN (phosphatase and tensin homolog deleted on chromosome 10) tumor suppressor gene codifies a lipid inositol 3'-phosphatase that negatively regulates cell survival mediated by the phosphatidyl inositol 3' kinase (PIP3 kinase)--protein kinase B/Akt signaling pathway. Recently, PIP3-kinase was involved in axon polarization, but PTEN functions in dendrites are uncertain. Using amino-terminal antibodies against the catalytic domain, we found a 34 kDa fragment of PTEN protein detected only in mouse brain tissue, present in neuron dendrites and spines of cerebral cortex, cerebellum, hippocampus and olfactory bulb. The PTEN-fragment reaches the synaptic fraction with a positive temporal correlation with synaptic stabilization in postnatal cerebellum and brain. In the weaver mutant mice, PTEN was absent only in the Purkinje cells dendrites that cannot receive the granule cells synaptic input. Furthermore, the activated p Akt/PKB was present in axons but not in dendrites of mature neuron cells. P-Akt was also altered by the weaver mutation maintaining the inverse correlation with the PTEN-fragment in Purkinje cell dendrites. In contrast, the expression of this fragment was not affected by the staggerer mutation. Together, these results suggest that synaptogenesis is a necessary process for polarization in PIP3 pathway mediated by the PTEN catalytic-fragment into dendrites of CNS neurons. PMID- 15337314 TI - Angiotensin II attenuates NMDA receptor-mediated neuronal cell death and prevents the associated reduction in Bcl-2 expression. AB - While angiotensin II (Ang II) plays a major role in the regulation of blood pressure, fluid homeostasis and neuroendocrine function, recent studies have also implicated the peptide hormone in cell growth, differentiation and apoptosis. In support of this, we have previously demonstrated that Ang II attenuates N-methyl D-aspartate (NMDA) receptor signaling [Molec. Brain Res. 48 (1997) 197]. To further examine the modulatory role of Ang II on NMDA receptor function, we investigated the effect of angiotensin receptor (AT) activation on NMDA-mediated cell death and the accompanying decrease in Bcl-2 expression. The viability of differentiated N1E-115 and NG108-15 neuronal cell lines was reduced following exposure to NMDA in a dose-dependent manner. MTT analysis (mitochondrial integrity) revealed a decrease in cell survival of 49.4+/-12.3% in NG108 cells and 79.9+/-6.8% in N1E cells following treatment with 10 mM NMDA for 20 h. Cytotoxicity in N1E cells was inhibited by the noncompetitive NMDA receptor antagonist, MK-801. Further, NMDA receptor-mediated cell death in NG108 cells was attenuated by treatment with Ang II. The Ang II effect was inhibited by both AT1 and AT2 receptor antagonists, losartan and PD123319, respectively, suggesting that both receptor subtypes may play a role in the survival effect of Ang II. Since it has been shown that activation of NMDA receptors alters the expression of Bcl-2 family proteins, Western blot analysis was performed in N1E cells to determine whether Ang II alters the NMDA-induced changes in Bcl-2 expression. A concentration-dependent decrease of intracellular Bcl-2 protein levels was observed following treatment with NMDA, and this reduction was inhibited by MK801. Addition of Ang II suppressed the NMDA receptor-mediated reduction in Bcl 2. The Ang II effect on NMDA-mediated changes in Bcl-2 levels was blocked by PD123319, but was not significantly changed by losartan, suggesting AT2 receptor specificity. Taken together, these results suggest that Ang II attenuates NMDA receptor-mediated neurotoxicity and that this effect may be due, in part, to an alteration in Bcl-2 expression. PMID- 15337315 TI - Neuroprotective effect of tacrolimus (FK506) on ischemic brain damage following permanent focal cerebral ischemia in the rat. AB - We investigated the neuroprotective effect of tacrolimus (FK506) on the ischemic cell death with respect to cytochrome c translocation and DNA fragmentation, which are pivotal events in the necrotic and apoptotic signaling pathway, using permanent focal cerebral ischemia in rats. Immunohistochemically, cytochrome c was observed in the cytoplasm as early as 1 h after middle cerebral artery (MCA) occlusion in the infarcted hemisphere. Cytosolic release of cytochrome c after MCA occlusion was also confirmed by Western blot analysis and enzyme immunoassay. Terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL) showed DNA fragmentation evolving in the ipsilateral cortex and the caudate putamen after 3 and 6 h, respectively, following MCA occlusion. Tacrolimus (1 mg/kg, i.v.), administered immediately after MCA occlusion, significantly attenuated the release of cytochrome c in the ischemic region, the number of TUNEL-positive cells in the ischemic penumbra zone, and the size of cortical ischemic lesions. This study demonstrated that tacrolimus ameliorated the accumulation of cytochrome c in the cytosol and the increase of TUNEL-positive cells induced by cerebral ischemia, indicating that the neuroprotective action of tacrolimus on ischemic brain injury caused by permanent focal cerebral ischemia could partially be attributed to the attenuation of the activation of the apoptotic execution machinery. PMID- 15337316 TI - Differential expression of Huntington's disease gene (IT15) mRNA in developing rat brain. AB - Huntington's disease (HD) is an autosomal dominant inheritance neurological disorder associated with CAG repeats expansions within a widely distributed gene that causes selective neuronal death. The gene is essential for normal development and has been suggested for its role in the development of basal ganglia. To understand its normal function during growth and development, we determined the expression patterns for the gene responsible for HD (IT15) mRNA by Northern blot analysis in the developing and adult rat brain. In adult rat brains, IT15 transcripts exist as two isoforms of 10 and 12.5 kb each, which display distinct expression patterns. IT15 mRNA expression is already highly expressed within 1 day of birth. Expression signals for IT15 were first detected at embryonic day 16 or 17 during prenatal development. IT15 transcript peaks leveled around day 20 postnatally and thereafter decreased to levels typically found in adults. In the adult rat brain, mRNA expression was highest in the cerebellum followed by the cortex, striatum, hippocampus and olfactory lobe. In the medulla and the spinal cord, IT15 was weakly expressed in comparison to the other regions studied. Thus, the distinct expression patterns provide a basis for its functional significance during development. These results also suggest that the degree of IT15 mRNA expression is related to the neuronal population in different brain regions. PMID- 15337317 TI - Free radical formation in cerebral cortical astrocytes in culture induced by methylmercury. AB - Oxidative stress has been implicated in neurotoxic damage associated with various metals, including methylmercury (MeHg). Although the mechanism(s) of MeHg-induced neurotoxicity remains unclear, evidence supports a mediatory role for astrocytes, a cell type that preferentially accumulates MeHg. Using scanning confocal microscopy (LSCM), the present study was undertaken to examine the role of astrocytes as the site of reactive oxygen species (ROS). Three redox-sensitive fluorescent probes were used for ROS analysis, (a) CM-H2DCFDA (chloromethyl derivative of dichlorodihydrofluorescein diacetate), a probe for intracellular hydrogen peroxide (H2O2); (b) hydroethidine (HETH), a probe for superoxide anion (*O2-), and (c) CM-H2XRos (chloromethyl derivative of dihydro X-rosamine), and a probe that is selective for mitochondrial reactive oxygen intermediates. Astrocytes were treated with 10 microM MeHg for 30 min, following which the various fluorescent probes were added; 20 min later LSCM images were collected. Astrocytes loaded with CM-H2DCFDA and HE demonstrated a significant MeHg-induced increase in fluorescence intensity indicative of increased intracellular H2O2 and *O2-, respectively. Similar results were obtained with the mitotracker dye, CM H2XRos. Additionally, exposure of astrocytes for 24 h to 100 microM buthionine-L sulfoxane (BSO), a glutathione (GSH) synthesis inhibitor, caused a significant increase in ROS formation. Furthermore, BSO pretreatment significantly enhanced the MeHg-induced formation of *O2-, indicating an important role for GSH in the maintenance of optimal cellular redox status. Time-course experiments performed in the simultaneous presence of CM-H2XRos and CM-H2DCFDA demonstrated that the MeHg-induced CM-H2XRos fluorescence changes preceded those of CM-H2DCFDA, suggesting that the mitochondria represent an early primary site for ROS formation. Taken together, these studies illustrate that MeHg induces the generation of astrocyte-derived ROS and support a role for astrocytic ROS in MeHg associated neurotoxic damage. PMID- 15337318 TI - A novel basic helix-loop-helix (bHLH) transcriptional repressor, NeuroAB, expressed in bipolar and amacrine cells in the chick retina. AB - Basic helix-loop-helix (bHLH) transcription factors are implicated in cell fate determination and differentiation in neurogenesis. We identified a novel chick bHLH transcription factor, NeuroAB. A phylogenetic tree prepared from bHLH sequences suggested that NeuroAB belongs to the BETA3 group in the Atonal-related protein family (ARPs). In situ hybridization and immunostaining indicated that NeuroAB is expressed predominantly in postmitotic bipolar cells and GABAergic amacrine cells in the retina. Reporter and DNA pull down assays indicated that NeuroAB functions as a transcriptional repressor by binding to the E-box sequence, and its activity is modulated by phosphorylation at a specific serine residue that fits the consensus phosphorylation site for glycogen synthase kinase 3beta (GSK3beta). Since members of the BETA3 group possess this consensus site, it is suggested that their activities are commonly regulated by GSK3beta or other kinases bearing the same substrate specificity. We found that the expression of GSK3beta is spatially and temporally regulated in the developing retina; its strong expression was observed in ganglion cells from E8 and a subset of amacrine cells from E12. These findings suggest that NeuroAB is involved in the maturation and maintenance of bipolar cells and GABAergic amacrine cells and regulation by GSK3beta plays an important role in retinogenesis. PMID- 15337319 TI - Neuronal cell lines transfected with the dopamine D2 receptor gene promoter as a model for studying the effects of antidepressant drugs. AB - The present study investigated the effect of three antidepressant drugs (ADs), desipramine (DMI, a noradrenaline reuptake inhibitor), citalopram (CIT, a selective serotonin reuptake inhibitor) and mianserin (MIA, thought to act as an antagonist of pre-synaptic alpha2 adrenoceptor) on the transcriptional activity of the dopamine D2 receptor gene promoter. The fragment of dopamine D2 receptor gene promoter (-850 to +133) was subcloned into pGL3 vector (Promega), which has an insert coding for luciferase used as a reporter gene. Such construct (pGL3 D2R) was used to transiently transfect the neuroblastoma cell lines, Neuro 2a, SH SY5Y and NB41A3, which endogenously express the dopamine D2 receptor protein. The obtained results indicate that transcriptional activity of dopamine D2 receptor gene promoter was dose-dependently increased by retinoic acid, forskolin, rolipram and phorbol 12 myristate 13-acetate, as well as by DMI, CIT and MIA. In the Neuro 2a cells, the most significant increase was observed after the ADs were present in the incubation medium at a doses of 0.1-1 microM for 72 h. In the SH SY5Y cells, the significant increase in the transcriptional activity of D2 receptor gene promoter was observed already after 24-h exposure to DMI. Incubation of the Neuro 2a cells in the presence of forskolin (1 microM) or rolipram (50 microM) (but not phorbol 12-myristate 13-acetate at 0.1 microM) in combination with DMI resulted in the further increase in transcriptional activity of the studied promoter, indicating the involvement of protein kinase A pathway in these effects. PMID- 15337320 TI - Changes in the gene and protein expression of K(ATP) channel subunits in the hippocampus of rats subjected to picrotoxin-induced kindling. AB - ATP-sensitive K+ (KATP) channels couple the intracellular metabolic state to electrical activity, which is important in the control of neuronal excitability and seizure propagation. In this study, we investigated the changes in the gene and protein expression of KATP channel subunits in the brain of picrotoxin (PTX) kindled rats, which were daily administered with a subconvulsant dose of PTX for 20 days. At 14 days after the last administration of PTX, kindled rats were retreated with PTX and killed by decapitation at 12 h, 1 and 3 days, as well as retreated with vehicle and killed at 0 h after starting the retreatment. The abundance of Kir6.1, Kir6.2, SUR1 and SUR2A/B mRNAs was evaluated by reverse transcription polymerase chain reaction (RT-PCR) using endogenous gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal control, and the level of Kir6.2 and SUR1 proteins was measured by Western blot. At 0 h, the brain showed decreasing expression of various subunit mRNAs, with the exception of the SUR2A mRNA. In contrast, from 12 h to 3 days, the amount of various subunit mRNAs was up-regulated dynamically, but SUR2A of which was not changed significantly both from cortical and hippocampal samples. Furthermore, we demonstrated that the levels of Kir6.1, Kir6.2, SUR1 and SUR2B mRNAs at 12 h and 3 days (excepting SUR1 at 3 days) from hippocampal samples, as well as Kir6.1 at 1 day and SUR1 at 3 days from cortical samples, were significantly higher than that detected at 0 h. In addition, low levels of Kir6.2 and SUR1 proteins were observed both from cortical and hippocampal samples at 0 h and also, from 12 h to 3 days, a marked up-modulation of Kir6.2 and SUR1 protein expressions especially from hippocampal samples was found. These results suggest that the PTX-induced changes in the KATP channels may play a key role in the induction and maintenance of kindling and the PTX-induced seizures might be important for the acute changes of KATP channels observed in kindled rat brain. PMID- 15337321 TI - Distribution and MK-801-induced expression of serine racemase mRNA in rat brain by real-time quantitative PCR. AB - We have used real-time quantitative PCR methods to evaluate the effect of the systemic administration of (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten 5,10-imine (MK-801; 0.4 mg/kg) on the gene expression of serine racemase in several brain areas of the rat. The levels of serine racemase mRNA in all the brain areas transiently increased after the administration. The present findings suggest that there is a link between the expression of serine racemase mRNA and the activity of the NMDA receptors. PMID- 15337322 TI - Expression of proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1beta in the brain during experimental group B streptococcal meningitis. AB - We performed mRNA in situ hybridization for TNF-alpha and IL-1beta from infant rats with group B streptococcal meningitis. Induction of both cytokines was seen in the ependyma and the meninges at 4 h. Both cytokines were expressed in the brain parenchyma at 12 h. Induction of IL-1beta mRNA was seen in vessels within the brain cortex. Neutrophilic infiltrate at all time points examined was minimal and could not account for the observed cytokine expression. PMID- 15337323 TI - Dysthymia in later life: a study in the community. AB - BACKGROUND: Dysthymia (DD) may be thought of as depression associated with personality disorder, a phase in the pleomorphic natural history of unipolar depression or a result of exposure to chronic physical illness. Prevalence, clinical features, risk factors and prognosis may change with AGE. METHOD: Large (n=3056) representative sample of elderly (55-85) in the Netherlands. Two-stage screen procedure to identify elderly with DD. The Center for Epidemiologic Studies Depression scale (CES-D) was used as a screen and the Diagnostic Interview Schedule (DIS) to diagnose DD. Data on 277 depressed elderly were available to assess the 6-year prognosis of DD. RESULTS: The prevalence of DD (4.61%) was higher in women and declined with age. The symptom profiles of DD and MDD were very similar. Those with DD were very likely to have had MDD earlier in life (44% in pure DD and 80% in those with double depression). The average age at onset (31 years) was earlier than in MDD (53 years). Environmental and personal vulnerability dominated the risk-factors. The prognosis was unfavourable in most cases. LIMITATIONS: Considerable attrition and retrospective data on age at onset and previous histories of depression. CONCLUSION: Although the prevalence declines with age, DD remains common in later life. Many cases arise later than is often thought and clinical features intertwine with those of MDD in the course of life. Given the unfavourable prognosis, provision of effective treatment is warranted. PMID- 15337324 TI - Immune parameters in patients with anxiety or depression during psychotherapy. AB - BACKGROUND: Numerous studies have described distinctive immunological findings in patients with depression. In contrast, only very little is known about the possible influence of anxiety disorders on the immune system. It is also unknown whether treatment with psychotherapy alone has any influence on immunological variations in patients with psychiatric disorders. METHODS: We measured immunological and psychological parameters in patients with minor depression (N=10) or anxiety disorder (N=13) over an 8-week course of inpatient psychotherapy. Data for patients and a group of healthy controls (N=11) were recorded three times in 4-week intervals. A FACS analysis revealed the composition of lymphocyte subpopulations. The production of reactive oxygen species (ROS) by phagocytes was analyzed using lucigenin-enhanced chemiluminescence. RESULTS: On admission, patients with anxiety disorder showed a markedly elevated ratio of CD4(+) (T helper) versus CD8(+) (T suppressor/cytotoxic) lymphocytes compared to healthy controls (P<0.001) and minor depressives (P<0.01). The increased ratio in patients with anxiety disorder could mainly be attributed to a reduced count in CD8(+) T cells compared to healthy controls (P<0.01) and depressives (P<0.05). There were no differences between patients with depression and healthy controls with respect to the CD4(+)/CD8(+) ratio. We did not observe any differences in the production of ROS by phagocytes in patients compared to healthy controls. The CD4(+)/CD8(+) ratio remained elevated in patients with anxiety disorders during the following 8 weeks. There were no significant changes in this parameter over the course of the inpatient treatment. LIMITATIONS: As a pilot study on the immune status in patients with anxiety disorders, the study's main limitation is the relatively low number of patients observed. CONCLUSIONS: In this study we demonstrated for the first time marked immunological changes in patients with anxiety disorders. In addition, our results provide preliminary evidence that these immunological variations are not reversible by an 8-week course of inpatient psychotherapy alone. PMID- 15337325 TI - Family caregiving in depression: impact on caregivers' daily life, distress, and help seeking. AB - BACKGROUND: Attention to caregiving consequences has been mainly restricted to schizophrenia, although studies suggest that relatives of depressed patients also experience considerable distress. These studies, however, were conducted on small samples or with nonvalidated instruments. In our study, the caregiving consequences of 260 spouses and relatives of depressed patients were assessed with a well-validated 31-item questionnaire, the Involvement Evaluation Questionnaire (IEQ). METHODS: The IEQ was mailed to spouses and relatives of patients with major depression, dysthymic disorder, or other depressive disorders. Other instruments used were the Ways of Coping Checklist (WCC), a Dutch Social Support Questionnaire, and the Zung Self-rating Depression Scale (SDS). RESULTS: About 25-50% of caregivers worried about the patient's general health, treatment, safety, and future. They had to urge the patient to undertake activities, or took over tasks. There were relational strains, and they felt burdened, especially when the patient was in an acute phase. Caregivers often felt distressed and had to visit a (mental) health practitioner. Also, children were affected; caregivers reported high levels of difficult behavior, loss of appetite, sleeplessness, less playing, and less attention at school. Caregiving consequences occur less often than in schizophrenia, but the patterns are quite comparable. CONCLUSIONS: Caregiving consequences in depression occur frequently and cause distress in caregivers and patient's children. Attention should be paid to support relatives and spouses of depressed patients. Special attention should be paid to patient's children. PMID- 15337326 TI - Quality of life assessment in patients with bipolar disorder treated with olanzapine added to lithium or valproic acid. AB - BACKGROUND: The objective of this study was to determine the clinical and quality of life outcomes associated with adjunctive treatment of olanzapine added to either lithium or valproic acid/divalproex sodium in patients with bipolar disorder. METHODS: Patients with bipolar I disorder, were randomized to receive either olanzapine (5-20 mg) added to mood stabilizer therapy (n=224), or placebo added to mood stabilizer therapy (n=112) for 6 weeks. Changes in clinical outcomes over 6 weeks were measured by the Young Mania Rating Scale (Y-MRS) and the Hamilton Rating Scale for Depression (HAM-D). Quality of life was measured by the Lehman Brief Quality of Life Interview (QLI). RESULTS: Patients treated with olanzapine added to mood stabilizers, experienced significantly greater mean clinical improvements from baseline on both the Y-MRS and the HAM-D compared to those treated with placebo added to mood stabilizers. Over 6 weeks, patients treated with olanzapine added to mood stabilizers had significantly greater mean improvements from baseline on five of the nine subjective scales on the QLI, compared to patients treated with placebo added to mood stabilizers. Changes in scores on the subjective scales of the QLI were more strongly correlated to changes in depressive symptomatology measured by the HAM-D, than to changes in symptoms of mania measured by the Y-MRS. CONCLUSION: The results of this study demonstrate that patients receiving adjunctive treatment have significantly greater improvements in both clinical and quality of life outcomes compared to monotherapy with mood stabilizers. PMID- 15337327 TI - Determinants of persistence of major depressive episodes in the general population. Results from the Netherlands Mental Health Survey and Incidence Study (NEMESIS). AB - BACKGROUND: Data on determinants of persistence of major depressive episodes (MDE) are inconsistent due to methodological shortcomings of the studies involved. AIMS: To examine determinants of persistence of MDE in subjects from the general population (N=250) with new episodes of DSM-III-R major depression. METHOD: The Netherlands Mental Health Survey and Incidence Study is a prospective epidemiologic survey in the adult population (N=7076), using the Composite International Diagnostic Interview (CIDI). A broad range of potential determinants was assessed. RESULTS: Determinants of persistence were severity of the index episode, longer duration of previous episodes, (chronic) physical illness and lack of social support. A recurrent episode predicted shorter duration. LIMITATIONS: Follow-up was only 2 years and duration of episode was retrospectively assessed. CONCLUSIONS: Just as in clinical populations, illness related factors seem to be the strongest predictors of persistence of MDE. A thorough assessment of each depressed patient on the predictors of persistence is advisable. PMID- 15337328 TI - Selecting key variables for depression prevention: results from a population based prospective epidemiological study. AB - BACKGROUND: Given the existing economic constraints, prevention of depression has to be carefully targeted, and applied where it is likely to yield the highest possible health benefits at the lowest possible cost. AIM: To identify those risk factors of depression that have the greatest use potential from the perspective of prevention. METHOD: Data were derived from a population-based prospective cohort of 4664 adults who had never experienced a depression. Their health status was re-examined after 1 year. Incidence rate ratios (IRR), population attributable risks (PAR) and numbers needed to be treated (NNT) were calculated to create a hierarchy of risk factors. RESULTS: Selecting high-risk groups with migraine, abdominal and respiratory complaints and markers of vulnerability or childhood trauma appears to be an indicated strategy. CONCLUSIONS: This study illustrates how epidemiology can contribute towards setting a Research and Development agenda for primary prevention of depression. PMID- 15337329 TI - A comparison of seasonal variation between suicide deaths and attempts in Hong Kong SAR. AB - BACKGROUND: Seasonal variations in suicide deaths and attempted suicides have been reported for many countries. Recent research has suggested that seasonal patterns of suicide deaths have diminished considerably. The seasonality pattern between attempts and completed ones are not the same. This paper is the first one to examine seasonal variation between suicide deaths and attempts simultaneously. METHOD: Census and Statistics Department and the Hospital Authority provided mortality and morbidity data on suicides for the period 1997-2001 in Hong Kong SAR, respectively. Seasonal patterns of suicide deaths and attempts were examined by a harmonic analysis and a non-parametric chi-square test. RESULTS: A significant seasonal variation was detected in suicide attempts with a markedly bi-seasonal pattern was found amongst females with a peak in May and October; only a cyclic pattern was observed for males with a peak in summer. The female attempters who used non-violent suicide methods contributed to the second peak in October. On the other hand, there was no evidence of significant differences in the seasonal distribution among the suicide deaths for males and females. Only a mild pattern was found amongst females involving in violent suicide deaths, while others can be treated as a random event and no significantly pattern was observed in our study. Overall, a significant difference in seasonal variation existed between suicide deaths and attempts (p-value<0.01). CONCLUSIONS: This study reinforces the findings that seasonal variation in suicide appears to be diminishing. Also, suicide attempters and completed suicides in Hong Kong seem to arise from two non-identical but overlapping groups in the community. PMID- 15337330 TI - Schneiderian first rank symptoms predict poor outcome within first episode manic psychosis. AB - BACKGROUND: The validity of a sub-classification of affective psychosis according to the mood congruence of psychotic features has been questioned in the literature. While some authors have found a correlation between such symptoms and outcome, their predictive value was rather limited in these studies. METHOD: Prospective study of 108 subjects presenting with a first DSM-III-R manic episode with psychotic features to determine the frequency of different types of psychotic symptoms and to measure the predictive utility of mood incongruent psychotic symptoms (MIPS) and first-rank Schneiderian symptoms (FRSS) during the first episode for a 12-month outcome. Outcome was measured by the level of positive, negative, depressive symptoms, and psychosocial functioning. Duration of affective and psychotic symptoms was also assessed. RESULTS: Patients presented with a wide variety of psychotic symptoms. The presence of MIPS at baseline was significantly correlated with a longer persistence of psychotic symptoms, but not with poorer outcome at 12 months. By contrast, the presence of FRSS at baseline was significantly associated with earlier onset of psychosis as well as increased severity of negative symptoms and poorer psychosocial functioning after 12 months. CONCLUSION: The presence of FRSS during a first manic episode with psychotic features identifies a sub-group of patients with more severe presentation and poorer short-term outcome. These results question the prognostic utility of MIPS. LIMITATIONS: Despite the relatively large number of subjects compared with other studies, the statistical power to detect all but large effect sizes is limited by the sample size. PMID- 15337331 TI - No effect of vitamin B-12 treatment on cognitive function and depression: a randomized placebo controlled study. AB - BACKGROUND: Associations between vitamin B-12 deficiency and impaired cognitive function and depression have been reported. METHODS: A randomized placebo controlled study including 140 individuals with an increased plasma methylmalonic acid (0.40-2.00 micromol/l) not previously treated with vitamin B-12. Cognitive function was assessed by the Cambridge Cognitive Examination (CAMCOG), Mini Mental State Examination (MMSE), and a 12-words learning test. Symptoms of depression were evaluated by the Major Depression Inventory. The main outcome measure was change in cognitive function and depression score from baseline to follow-up 3 months later. RESULTS: At baseline 78 (56%) individuals had cognitive impairment judged from the CAMCOG score and 40 (29%) according to the MMSE; 18 (13%) individuals had symptoms of depression. No improvement was found in cognitive function comparing the treatment and placebo group (total CAMCOG score: P = 0.43), nor among individuals with only slightly impaired cognitive function (n = 44, total CAMCOG score: P = 0.42). The treatment group did not improve in depression score as compared to the placebo group (P = 0.18). LIMITATIONS: The duration of impaired cognitive function was unknown. CONCLUSIONS: A high proportion of individuals with an increased plasma methylmalonic acid had impaired cognitive function, and a rather high prevalence of depression was observed. However, vitamin B-12 treatment did not improve cognitive function or symptoms of depression within the 3-months study period. PMID- 15337332 TI - Failure to replicate the effects of gender and season on the length of hospitalisation in unipolar depressives. AB - BACKGROUND: Kecskes et al. [J. Affect. Dis. 73 (2003) 279] reported a possible effect of season and gender on the length of hospital stay among inpatients with a unipolar depression. METHODS: The effects of season and gender upon the length of stay and clinical outcomes among 1283 inpatient admissions with unipolar depression diagnosed according to ICD-10 were examined. RESULTS: Males had shorter length of stay but no better outcomes. There was no evidence of shorter lengths of stay for either gender in summer. CONCLUSIONS: The present study failed to replicate the effect of season and gender on the length of stay in hospital among inpatients with unipolar major depression. LIMITATIONS: The clinical outcomes were based upon a single item from the Health of the Nation Outcome Scales (HoNOS). PMID- 15337333 TI - Subtypes of manic episodes according to ICD-10-prediction of time to remission and risk of relapse. AB - BACKGROUND: The long-term predictive ability of the ICD-10 subtypes of hypomania, mania without psychosis and mania with psychosis has not been investigated. METHODS: All patients who got a diagnosis of a single hypomanic episode, single manic episode without psychosis or single manic episode with psychosis in a period from 1994 to 1999 at the first discharge ever in Denmark were identified. The time to discharge from first admission and the risk of relapse leading to readmission were compared for the three groups of patients. RESULTS: Totally, 41 patients with a hypomanic episode, 149 patients with a manic episode without psychotic symptoms and 202 patients with a manic episode with psychotic symptoms at first discharge ever were identified. Patients with mania and psychotic symptoms were admitted longer than patients with mania without psychosis, and patients with mania without psychosis were admitted longer than patients with hypomania. No differences were found in the risk of relapse leading to readmission between the three groups of patients. LIMITATIONS: The results apply to hospitalised patients only. CONCLUSIONS: The ICD-10 subclassification of manic episodes does only partially predict long-term outcome. PMID- 15337334 TI - Concordance therapy: an intervention to help older people take antidepressants. AB - BACKGROUND: Older people often do not adhere fully to antidepressant medication. Compliance Therapy improves adherence with antipsychotic medication. OBJECTIVE: To adapt Compliance Therapy for use in older depressed patients, to pilot this 'Concordance Therapy' for feasibility and acceptability and to gain preliminary indications of its efficacy. METHODS: Randomised controlled trial (RCT). SETTING: Psychiatric services for older people in North London and Essex. PARTICIPANTS: A total of 19 older depressed patients. INTERVENTION: 10 patients received Concordance Therapy over 3-4 sessions. CONTROL: 9 patients received treatment as usual. MAIN OUTCOME MEASURE: medication adherence at 1 month. SECONDARY OUTCOME MEASURES: medication adherence at 3 months; depression severity, beliefs about medication, quality of life at 1 and 3 months; patient feedback about the therapy. RESULTS: The therapy was acceptable to patients. INTERVENTION patients were more likely to take antidepressants, had a higher quality of life, had less depressive symptomatology and were less likely to be cases of depression at 1 month. Beliefs around antidepressants at 1 month were more positive in the intervention group but this was not the case for medication in general. LIMITATIONS: As a pilot, patient numbers were small and the findings did not reach statistical significance. Three patients (1 intervention, 2 control) were in hospital and therefore offered medication at follow-up. CONCLUSION: Concordance Therapy for older people prescribed antidepressants is acceptable and feasible and shows sufficient promise of efficacy to justify an adequately powered RCT. PMID- 15337335 TI - Significant influence of smoking on the mood in volunteers who had the common cold. PMID- 15337336 TI - A review of licensed viral vaccines, some of their safety concerns, and the advances in the development of investigational viral vaccines. AB - Viral vaccines could be considered among the most important medical achievements of the 20th century. They have prevented much suffering and saved many lives. Although some curative antiviral drugs exist, we desperately depend on efforts by academic, governmental and industrial scientists in the advancement of viral vaccines in the prevention and control of infectious diseases. In the next decade, we hope to see advancement in the development of current and investigational viral vaccines against childhood and adult infections. In this article, we will review the licensed viral vaccines, some of their safety concerns, and the advances in the development of investigational viral vaccines. PMID- 15337337 TI - Heroes of SARS: professional roles and ethics of health care workers. AB - OBJECTIVES: To examine the professional moral duty of health care workers (HCWs) in the outbreak of severe acute respiratory syndrome (SARS) in 2003. METHODS: Descriptive discussion of media reports, analysis of ethical principles and political decisions discussed in the outbreak, with particular emphasis on the events in mainland China and Taiwan. RESULTS: There were differences in the way that Taiwan and mainland China responded to the SARS epidemic, however, both employed techniques of hospital quarantine. After early policy mistakes in both countries HCWs were called heroes. The label 'hero' may not be appropriate for the average HCW when faced with the SARS epidemic, although a number of self-less acts can be found. The label was also politically convenient. CONCLUSIONS: A middle ground for reasonable expectations from HCW when treating diseases that have serious risk of infection should be expected. While all should act according to the ethic of beneficence not all persons should be expected to be martyrs for society. PMID- 15337338 TI - Apoptosis of naive CD4+ T-cells from HIV-infected patients with poor immune response to HAART is enhanced in vitro by steroid. AB - OBJECTIVE: Because the absence of immune restoration in HIV-infected patients efficiently treated by highly active antiretroviral therapy (HAART) may be due to excessive immune activation, we prospectively studied the effect of hydrocortisone on T-cell apoptosis in a cohort of patients with satisfactory virologic response. METHODS: Apoptosis of T-cell subsets including naive CD45RA(+)CD4+ T-cells was determined at baseline and at months 1 and 3 after initiation of HAART. A satisfactory immune response was defined as an increase >100/microL CD4+ T-cells at month 3 compared to baseline. RESULTS: Twenty out of 63 patients showed undetectable viral load at month 3, among whom eight exhibited a satisfactory immune response. Down-regulation spontaneous CD4+T-cell apoptosis was significant in the group of patients with a satisfactory immune response compared to the other patients. However, hydrocortisone up-regulated apoptosis of naive CD4+ CD45RA+ T-cells, specifically in group of patients with poor immune response, whatever the time point considered: percentage of apoptotic CD4 T-cells was 16+/-16% without hydrocortisone and 22+/-22% with hydrocortisone at month 1, and respectively, 10+/-9 and 17+/-15% at month 3 (P < 0.05) Hydrocortisone had no impact on CD8+ T-cell apoptosis, whatever the considered group. CONCLUSION: Our results suggest to not use steroid therapy as adjuvant immunotherapy in patients with less than optimal immunologic response to HAART. PMID- 15337339 TI - Respiratory viruses seasonality in children under five years of age in Buenos Aires, Argentina: a five-year analysis. AB - OBJECTIVES: During the winter months there is a remarkable increase in paediatric hospitalisations due to viral acute lower respiratory infections (ALRI). We aimed to perform a five-year retrospective analysis (1998-2002) of ALRI viral aetiology in children under 5 years of age admitted to public hospitals in Buenos Aires city to evaluate its seasonality. MATERIALS AND METHODS: Nasopharyngeal aspirates (NPA) were analysed by indirect immunofluorescence to determine viral aetiology. A Spearman's rank correlation test between meteorological parameters and viral frequencies was performed. RESULTS: Viruses were identified in 6083 (32.8%) of 18,561 NPA tested. Among the former 4796 (78.8%) were RSV, 508 (8.3%) IA, 473 (7.8%) AV, 293 (4.8%) PIV and 13 (0.2%) IB. RSV and IA peaked during the coldest and dampest months, whereas PIV did so in early spring and AV lasted throughout the year. For RSV and IA an inverse correlation with mean monthly temperature (r = -0.9 and r = -0.87, respectively, p<0.0001) and solar UVB radiation (r = -0.92 and r = -0.80, respectively, p<0.0001) was detected, while it was positive when relative humidity was considered (r = 0.6, p<0.0001 and r = 0.47, p=0.0068, respectively). CONCLUSIONS: This study highlights the seasonal variation of ALRI and allows the implementation of adequate healthcare strategies and practice guidelines. PMID- 15337340 TI - Hepatocyte growth factor (HGF) in patients with hepatitis B and meningitis. AB - OBJECTIVES: The present study investigates serum hepatocyte growth factor (HGF) levels in patients with acute and chronic hepatitis B and the relation between these levels and intrahepatic inflammatory markers of the liver and fibrosis, as well as the cerebrospinal fluid (CSF) HGF levels in patients with meningitis and the relation between these levels and CSF findings. To our knowledge this is the first study regarding CSF HGF levels in tuberculous meningitis. PATIENTS AND METHODS: The study consisted of 35 patients with chronic hepatitis B (HbeAg and HBV-DNA positive), 20 with acute hepatitis B, 20 with acute bacterial meningitis and 15 having tuberculous meningitis. HGF levels in the serum and CSF samples were measured by using the ELISA method. RESULTS: The mean serum HGF levels in acute hepatitis B group were found statistically significantly higher than those in the control group and chronic hepatitis B group (p<0.0001). It was established that serum HGF levels in patients with chronic hepatitis B were significantly correlated with serum alanine aminotransferase (ALT) and HBV-DNA levels (r: 0.816, 0.951; p<0.05, respectively). Similarly, serum HGF levels of patients with chronic hepatitis B were correlated with fibrosis score and hepatic activity index of the liver histopathology (r: 0.750, 0.459; p<0.05, respectively). The mean CSF HGF levels of patients with acute bacterial meningitis and tuberculous meningitis were higher than those in the control group (p<0.05). In addition, it was observed that mean CSF HGF levels in patients suffered from tuberculous meningitis were statistically significantly higher than those in acute bacterial meningitis (p<0.05). CONCLUSIONS: We suggest that serum HGF level in patients with chronic hepatitis B might reflect viral load, necro-inflammatory activity in the liver and the degree of structural progression. Our findings have demonstrated that tuberculous meningitis cause increased HGF concentrations in CSF. It is, therefore, suggested that examination of HGF levels in CSF may provide additional information in the differential diagnosis. PMID- 15337341 TI - Cryoglobulinemia and progression of fibrosis in chronic HCV infection: cause or effect? AB - OBJECTIVES: Mixed cryoglobulinemia (MC) is the most common extrahepatic manifestation of HCV infection. The aim of this study is to determine the prevalence of MC in HCV infected Greek patients and to identify if it is associated with liver histology or the mode of HCV transmission. METHODS: One hundred and twenty-six patients with chronic HCV infection were evaluated for the presence of serum cryoglobulins, autoantibodies and viral markers. One hundred and eighteen of them underwent liver biopsy and each specimen was evaluated according to the grading and staging system described by Ishak et al. RESULTS: Cryoglobulins were detected in 37/126 (29.4%) HCV patients and cryocrit values ranged between 0.5 and 6.5%. Only two patients presented clear clinical manifestations of MC. In patients with MC, a higher grading (6.40+/-2.06 vs. 5.27+/-2.55, p=0.013) and staging score (3.71+/-1.45 vs. 2.83+/-1.84, p=0.007) was noted in liver biopsy compared to those without MC. Logistic regression analysis identified staging score (OR, 1.33; CI, 1.06-1.66, p=0.015) as the only independent variable associated with cryoglobulinemia. Correlation between the presence of cryoglobulins and the mode of HCV transmission was not found. CONCLUSIONS: Greek patients with chronic HCV infection have high prevalence of cryoglobulinemia. A clear association between the presence of serum cryoglobulins and staging score of chronic hepatitis was found, with no difference in patients' age or the duration of infection. It is possible that cryoglobulinemia results in more rapid hepatic fibrosis in HCV infected patients. PMID- 15337342 TI - Incidence of pertussis in persons < or =15 years of age in Valencia, Spain: seroprevalence of antibodies to pertussis toxin (PT) in children, adolescents and adults. AB - PURPOSE: To determine the incidence of pertussis in persons < or =15 years in age in Valencia, Spain. To assess the prevalence of IgG antibodies to pertussis toxin (PT) in children, adolescents and adults. METHODS: Prospective study conducted at paediatric primary care centres. All persons < or =15 years in age presented with persistent cough were enrolled. Parents completed a brief questionnaire and immunization history was obtained from paediatrician records. A blood sample was obtained, for determination of IgG antibodies to Bordetella pertussis toxin (PT) by an ELISA method. A study confirmed-case was the presence of two conditions: (1) cough illness of > or =14 days duration; and (2) ELISA absorbance value of IgG to PT > or =2. Two subjects per clinical-case (same centre and range of age) and parents were asked to participate in the prevalence study. RESULTS: Sixty-one children < or =15 years in age presented with symptoms leading to a clinical diagnosis of pertussis were detected. Serological evidence of recent pertussis was found in five of these patients (incidence of 46.0/100,000 persons < or =15 years in age). Prevalence of antibodies to B. pertussis (> or =0.3) in children < or =15 years in age and adults was 39 and 33%, respectively. Only a minority of children, adolescents and adults had absorbance values indicative of immunity (> or =1). CONCLUSIONS: These incidence and seroprevalence results show that despite high immunization rates in infancy, B. pertussis is circulating in Spain. PMID- 15337343 TI - The effect of current management on morbidity and mortality in hospitalised adults with funguria. AB - BACKGROUND: To compare morbidity and mortality in inpatients with asymptomatic funguria between those treated and those observed for funguria. METHODS: Retrospective analyses were performed in 149 consecutive adult tertiary care inpatients with asymptomatic funguria. The primary endpoints were death, length of hospitalisation and progression to invasive fungal infection (IFI). RESULTS: Of the 149 subjects, 70% were female, 55% were >65 years, recent antibiotic and urinary catheter use occurred in >70%, diabetes in 32%, recent ICU admission in 29%, and concomitant bacteriuria in 28%. Forty-seven percent did not receive active intervention. Of the remainder, 46% were managed by controlling or eliminating risk factors for funguria or progression to IFI; fluconazole or amphotericin B were used to treat the other 54%. Fourteen percent died and 2.7 % progressed to IFI, with no significant difference between the treated versus observed groups for either endpoint (p>0.2). Median length of hospitalisation was significantly greater in the treated group (p<0.01); multivariate analysis demonstrated an exclusive relationship to the greater number of risk factors present in the treated group. CONCLUSION: Asymptomatic funguric patients who were managed with risk reduction and/or antifungal therapy were older, had more risk factors for funguria and subsequent progression to IFI, and had a longer hospital admission than those managed with observation. Treatment of asymptomatic funguria with risk reduction and/or antifungal therapy did not impact adult inpatient morbidity or mortality in this review; rather, the presence of multiple risk factors for funguria or IFI appeared to serve as a 'sickness indicator'. SUMMARY: In this study, we found that treatment for asymptomatic funguria in hospitalised adults did not impact morbidity or mortality. Rather, the presence of multiple risk factors for funguria or IFI correlated with a longer duration of hospitalisation, suggesting that funguria may be a 'sickness indicator', similar to bacteriuria in the elderly. PMID- 15337344 TI - Cryptococcus neoformans var. gattii meningitis in an HIV-positive patient: first observation in the United Kingdom. AB - We present a case of meningitis due to Cryptococcus neoformans var. gattii in an HIV-infected man recently returned from South Africa. We believe this is the first imported case of meningitis secondary to Cryptococcus neoformans var. gattii in an HIV-infected patient to be reported in the UK. PMID- 15337345 TI - Evaluation of the pollution of the surface waters of Greece from the priority compounds of list II, 76/464/EEC directive, and other toxic compounds. AB - The pollution of the surface waters of Greece from the priority compounds of 76/464/EEC Directive was evaluated. The occurrence of 92 toxic compounds, 64 of which belong to priority compounds of List II, candidates for List I, of 76/464/EEC Directive, was studied in surface waters and wastewater through the developed network of 62 sampling stations, which covers the whole Greek territory. The analytical determination was performed by Purge and Trap-Gas chromatography-Mass spectrometry for volatile and semivolatile organic compounds (VOCs), Gas Chromatography-Electron Capture Detection for organochlorine insecticides, Gas Chromatography-Nitrogen Phosphorous Detection for organophosphorous insecticides, High Performance Liquid Chromatography-Photodiode Array Detection for herbicides, and Electrothermal Atomic Absorption Spectrophotometry and Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES) for metals and the toluene extractable organotin compounds. The concentrations of VOCs and insecticides detected in the surface waters of Greece were very low, whereas the concentrations of herbicides and metals ranged generally at moderate levels. VOCs were detected almost exclusively in the rivers and very rarely in the lakes, while the frequency of occurrence of insecticides, herbicides and metals was similar for rivers and lakes. Water quality objectives (WQO) and emission limit values (ELV) have been laid down in national legal framework for a number of compounds detected in the samples, in order to safeguard the quality of surface waters from any future deterioration. PMID- 15337346 TI - Characterization of heavy metal particles embedded in tire dust. AB - Tire dust is a significant pollutant, especially as a source of zinc in the urban environment. This study characterizes the morphology and chemical composition of heavy metal particles embedded in tire dust and traffic-related materials (brake dust, yellow paint, and tire tread) as measured by a field emission scanning electron microscope equipped with an energy dispersive X-ray spectrometer (FESEM/EDX). In 60 samples of tire dust, we detected 2288 heavy metal particles, which we classified into four groups using cluster analysis according to the following typical elements: cluster 1: Fe, cluster 2: Cr/Pb, cluster 3: multiple elements (Ti, Cr, Fe, Cu, Zn, Sr, Y, Zr, Sn, Sb, Ba, La, Ce, Pb), cluster 4: ZnO. According to their morphologies and chemical compositions, the possible sources of each cluster were as follows: (1) brake dust (particles rich in Fe and with trace Cu, Sb, and Ba), (2) yellow paint (CrPbO(4) particles), (3) brake dust (particulate Ti, Fe, Cu, Sb, Zr, and Ba) and heavy minerals (Y, Zr, La, and Ce), (4) tire tread (zinc oxide). When the chemical composition of tire dust was compared to that of tire tread, the tire dust was found to have greater concentrations of heavy metal elements as well as mineral or asphalt pavement material characterized by Al, Si, and Ca. We conclude that tire dust consists not only of the debris from tire wear but also of assimilated heavy metal particles emitted from road traffic materials such as brake lining and road paint. PMID- 15337347 TI - Metal and metallothionein level in the heat-treated cytosol of gills of transplanted mussels Mytilus galloprovincialis Lmk. AB - A stock of mussels Mytilus galloprovincialis was transplanted over 1 year to four sites in a semi-enclosed bay in Croatia which is under the influence of various sources of pollution. The positive correlation of metal (Cd, Zn, Cu, Mn, Fe) and metallothionein (MT) tissue contents based on the analysis of heat-treated cytosol of gills with shell mass, as an indicator of mussel age, indicated to accumulation of metals, as well as the increase of MTs with mussel age. The principal component analysis (PCA) revealed that 74% of total variance of obtained results could be explained through two principal components. The first principal component was highly correlated with MT, Cd and Zn indices (metal or MT content/shell mass), and the second one with Cu, Mn, and Fe indices, as well as the gill index. High correlation of MTs with Zn and Cd is consistent with their affinity for binding to MTs. PMID- 15337348 TI - Health risk assessment of PCDD/F emissions from a hazardous and medical waste incinerator in Turkey. AB - A multimedia risk assessment procedure was conducted to determine the fate and transport of polychlorinated dibenzo-p-dioxin and polychlorinated dibenzofuran (PCDD/F) emissions from the Izmit Hazardous and Medical Waste Incinerator (IZAYDAS) in Turkey and their potential health risks. Congener concentrations in ambient air and their deposition rates were determined by an air dispersion model (ISCST3). Their transfer to some plant groups and animal tissues was predicted by food chain modeling. Exposure scenarios were produced based on three receptor groups (urban, semiurban, and rural) and five subgroups determined according to ages. Daily intakes of PCDD/Fs via exposure pathways were determined based on three different emission scenarios for each group and subgroup. Estimated incremental PCDD/F doses caused by the incinerator emissions in central-tendency scenario were computed as between 2.31 x 10(-6) and 0.008 pg TEQ kg(-1) bodyweight (bw) day(-1) on average for all the receptors other than infants in all the settings, while the range was 3.01 x 10(-5)-0.081 pg TEQ kg(-1) bw day( 1) for infants. Sensitivity analysis showed that the consumption of vegetal products and their locally grown fractions are the most significant parameters in the exposure to PCDD/Fs in the area. PMID- 15337349 TI - Statistical comparison and correlation of zinc and lead in estuarine sediments along the western coast of Mauritius. AB - Cr, Zn and Pb were quantified using atomic absorption spectrometry (AAS) from urban and rural estuarine sediments collected along the western coast of Mauritius during the period July 2002-January 2003. The mean concentration of Cr (225.4 mg kg(-1)), Zn (107.0 mg kg(-1)) and Pb (27.0 mg kg(-1)) in sediments along the six estuaries can be considered below those from contaminated estuarine sediments. Zn and Pb were significantly higher in urban sediments downstream St. Louis River compared to a rural estuary at Tamarin at 5% significance level. Zn was also significantly lower in Tamarin compared to Petite Riviere Noire estuary. Cr was however not significantly different in sediments from the six estuaries and could be considered of basaltic origin and were correlated to neither lead nor zinc. The concentration of Zn and Pb were greatest in the more urbanised upper and lower reaches of the St. Louis estuary during the period of study and were also significantly positively correlated in the estuarine sediments indicating that the cycling of Pb and Zn were linked indicating a common source for Pb and Zn. The potential sources of Pb and Zn in the urban estuarine sediments were considered to arise from road runoff causing significant quantities to be trapped. At present, compared to contaminant levels found in the UK and elsewhere, Mauritius still looks relatively uncontaminated but there is growing concern about heavy metal contamination in urban estuaries. There is therefore a need for long-term studies in estuarine management to involve integrated strategies in which water-sediment interactions in estuaries need to be considered and international standards for sediment quality developed. Such studies undertaken in the future would further the understanding of the biogeochemical processes of estuarine systems in Mauritius which could then be used in environmental development schemes and effective coastal zone management. PMID- 15337350 TI - Predicting CO2 and SO2 emissions in the Baltic States through reorganization of energy infrastructure. AB - The paper deals with predicting carbon dioxide and sulphur dioxide emissions generated by power production sector in the Baltic States in period up to year 2020. The economies of Lithuania, Latvia and Estonia are rapidly growing therefore forecast of emissions related with this occurrence becomes very important. The Ignalina Nuclear Power Plant (INPP), one of the largest in the world, is situated in the region. Two power production scenarios are modelled to investigate changes in power sector's emissions expected as the consequences of the coming closure of Ignalina NPP. Power market was assumed to be common for all three Baltic countries and was modelled by applying the Balmorel model. The planned closure of Ignalina NPP will bring restructuring of Lithuania power production sector and will change also power transmission between countries. Predictive identified the potential of investments for new modern power generation technologies. At the same time, modelling results show in both scenarios that CO(2) and SO(2) emissions from power production in the Baltic region will increase. The increment of emissions is discussed in the context of meeting requirements of UNFCCC Kyoto protocol and EC Directives. Despite of CO(2) emissions increase the Kyoto protocol's requirements may be expected. At the same time, SO(2) formation in Lithuania power sector may exceed the limits of the EU Council Directive 2001/80/EB therefore the additional measures to control SO(2) emissions have to be investigated. PMID- 15337351 TI - Evaluation of mixture effects in a crude extract of compost using the CALUX bioassay and HPLC fractionation. AB - Potential synergistic interactions between polycyclic aromatic hydrocarbons in a household sewage sludge compost extract were investigated using the Dioxin Responsive Chemical-Activated Luciferase gene eXpression (DR-CALUX) assay and reverse-phase high-performance liquid chromatography (RP-HPLC) fractionation. The biological activity of the crude extract was measured in vitro using the CALUX assay. The CALUX activity of the extract was as potent as 360-pg CALUX-TEQ (2,3,7,8-TCDD equivalent value) per g sample, this was 70 times above the WHO-TEQ value which was derived from chemical analyses of dioxins/furans and dioxin-like PCBs of the mixture. The CALUX activity pattern of the crude extract and the retention times of 26 polycyclic aromatic compounds (PACs), as determined by RP HPLC on an octadecylsilica column, suggested that the dioxin-like compounds with the log K(OW) (n-octanol/water partition coefficient) values corresponding to 6.0 7.0 contributed highly to the whole activity. The CALUX activity of the crude extract was three times the sum of the CALUX activities of the RP-HPLC separated fractions. Mixture effects were assessed by co-exposure of each HPLC fraction and 2,3,7,8-TCDD to the cells. The four concentration levels of added 2,3,7,8-TCDD corresponded to the TEQ value in the original compost sample. The experimental CALUX activity was higher than the predicted CALUX activity for some fractions. It was demonstrated that some compounds in the compost sample interacted synergistically with 2,3,7,8-TCDD in terms of dioxin-like activity. This finding points out the necessity for detailed investigation of synergistic effects in environmental samples. PMID- 15337352 TI - Relationship between fluorine in drinking water and dental health of residents in some large cities in China. AB - In this project, the relationship between fluorine content in drinking water and dental health of residents in some large cities in China was evaluated. The concentration of fluorine in tap water and in urine of local subjects of 28 cities and 4 high fluorine villages in China shows a strong positive correlation (r(2)=0.96, S.E.=0.9881). Our studies indicate that drinking water is the most important source of fluorine intake for Chinese people, and in more than 90% of urban cities, fluorine concentrations in drinking water are below levels recommended by the WHO (approximately 0.5-1.0 mg/l). A 1995 investigation by The National Committee on Oral Health of China (NCOH) shows the relationship between average number of decayed, missing and filled teeth (DMFT) of urban residents and fluorine concentration in drinking water to be negatively correlated but not forming a good linear relationship. Our results, together with the previous study, suggest that: (1) dental caries of the study population can be reduced by drinking water fluoridation and that (2) other factors such as economic level, weather, lifestyle, food habits, living condition, etc., of a city can also affect the incidence of dental caries that cannot be predicted by fluoridation alone. Research on the relation between index of fluorosis (IF) and the fluorine concentration in drinking water for the four high fluorine villages showed that the recommended concentration of fluorine in drinking water can protect from dental fluorosis. PMID- 15337353 TI - Emission modelling and validation of VOCs' source strengths in air-conditioned office premises. AB - An emission model for indoor volatile organic compounds (VOCs) based on mass balance considerations has been presented and validated under steady state conditions. Comparison were made for the measured and predicted concentrations of 37 selected VOCs and TVOC through a case intervention study on the filters of the ventilation system in a new commercial air-conditioned office building. The intervention involved replacing media filters with electronic and carbon filtration. TVOC and 37 compounds selected for their health and comfort impact, representation of major chemical classes that occur in indoor air and their utility as markers of pollution sources were studied. The concentration levels predicted by the model were compared with actual measurements. Twenty-five target compounds and the TVOC were adequately described by the model where the measured concentrations were in agreement with the predicted concentrations. Modeling of the remaining 12 compounds was found to be affected by the emission rates that were occupant related. PMID- 15337354 TI - Levels of elements in the surficial estuarine sediments of the Hugli River, northeast India and their environmental implications. AB - The present paper is the first document of a detailed geochemical and mineralogical study of muddy to sandy mud estuarine sediments of the Hugli River collected from five different sites along its course in the coastal areas of West Bengal, northeast India. The present work attempts to establish the status of distribution and environmental implications of 52 elements in the surficial estuarine sediments and their possible sources of derivation. The level of both metallic and non-metallic elements shows a wide range of variation all along the course of the estuary and can be attributed to their differential derivation from the source rocks and differential discharge of untreated effluents originating from industrial, agricultural, aquacultural as well as domestic sewage. The element contents, particularly the heavy metal content in the sediments, are the lowest in the upstream part of the estuary at Diamond Harbor, whereas, these are slightly higher in the intermediate stretch of the estuary at Haldia to highest in the mouth of estuary at Gangasagar. These changes indicate that the metals that are carried from upstream find their ultimate depositional sink at the delta mouth near Gangasagar, where almost all the elements showed elevated values. The majority of the elements have their highest concentrations at Canning, a site within the tidal channel network of the Hugli-Matla drainage basin away from the direct influence of the Hugli River. This site is severely contaminated with huge organic load from domestic sewage, aquaculture, intensive trawling activities and agricultural runoff. Moreover, the site suffers from heavy siltation load causing an almost moribund condition of Matla River at this point. Equi-dimensional quartzo-feldspathic mineral grains are consistent components in the siliciclastic composition of the sediments and their sizes at each station are controlled by respective hydrodynamic conditions. Various flaky minerals (mica, chlorite, hornblende) are also present but show inconsistent patterns of distribution. These flaky minerals remain in suspension for a long time and do not follow the usual law of settling of particles depending on the hydrodynamic regime prevailing in the area. Presented data will serve as a baseline against which future anthropogenic effects may be assessed. A comprehensive account of heavy metal content in sediments from different coastal regions of peninsular India has also been presented for purpose of comparison. PMID- 15337355 TI - Different methodologies to quantify uncertainties of air emissions. AB - Characterization of the uncertainty associated with air emission estimates is of critical importance especially in the compilation of air emission inventories. In this paper, two different theories are discussed and applied to evaluate air emissions uncertainty. In addition to numerical analysis, which is also recommended in the framework of the United Nation Convention on Climate Change guidelines with reference to Monte Carlo and Bootstrap simulation models, fuzzy analysis is also proposed. The methodologies are discussed and applied to an Italian example case study. Air concentration values are measured from two electric power plants: a coal plant, consisting of two boilers and a fuel oil plant, of four boilers; the pollutants considered are sulphur dioxide (SO(2)), nitrogen oxides (NO(X)), carbon monoxide (CO) and particulate matter (PM). Monte Carlo, Bootstrap and fuzzy methods have been applied to estimate uncertainty of these data. Regarding Monte Carlo, the most accurate results apply to Gaussian distributions; a good approximation is also observed for other distributions with almost regular features either positive asymmetrical or negative asymmetrical. Bootstrap, on the other hand, gives a good uncertainty estimation for irregular and asymmetrical distributions. The logic of fuzzy analysis, where data are represented as vague and indefinite in opposition to the traditional conception of neatness, certain classification and exactness of the data, follows a different description. In addition to randomness (stochastic variability) only, fuzzy theory deals with imprecision (vagueness) of data. Fuzzy variance of the data set was calculated; the results cannot be directly compared with empirical data but the overall performance of the theory is analysed. Fuzzy theory may appear more suitable for qualitative reasoning than for a quantitative estimation of uncertainty, but it suits well when little information and few measurements are available and when distributions of data are not properly known. PMID- 15337356 TI - Air pollution and the demand for hospital services: a review. AB - Time-series studies published since 1993 on the association between short-term changes in air quality and use of hospital services, including both inpatient and emergency room use, are reviewed. The use of nonparametric analysis, often incorporating generalized additive models (GAMs), has increased greatly since the early 1990s. There have also been three major multi-city studies, which together analyzed data from well over 100 cities in Europe and North America. Various air pollutants, especially ozone (O(3)), particulate matter (PM), nitrogen dioxide (NO(2)) and sulfur dioxide (SO(2)), were generally found to be significantly associated with increased use of hospital services. Ozone tends to have stronger effects in the summer during periods of higher concentrations. Several studies revealed synergistic effects between pollutants such as PM and SO(2). Overall, short-term exposure to air pollutants is found to be an important predictor of increased hospital and emergency room use around the world. PMID- 15337357 TI - The ecological effects of trichloroacetic acid in the environment. AB - Trichloroacetic acid (TCAA) is a member of the family of compounds known as chloroacetic acids, which includes mono-, di- and trichloroacetic acid. The significant property these compounds share is that they are all phytotoxic. TCAA once was widely used as a potent herbicide. However, long after TCAA's use as a herbicide was discontinued, its presence is still detected in the environment in various compartments. Methods for quantifying TCAA in aqueous and solid samples are summarized. Concentrations in various environmental compartments are presented, with a discussion of the possible formation of TCAA through natural processes. Concentrations of TCAA found to be toxic to aquatic and terrestrial organisms in laboratory and field studies were compiled and used to estimate risk quotients for soil and surface waters. TCAA levels in most water bodies not directly affected by point sources appear to be well below toxicity levels for the most sensitive aquatic organisms. Given the phytotoxicity of TCAA, aquatic plants and phytoplankton would be the aquatic species to monitor for potential effects. Given the concentrations of TCAA measured in various soils, there appears to be a risk to terrestrial organisms. Soil uptake of TCAA by plants has been shown to be rapid. Also, combined uptake of TCAA from soil and directly from the atmosphere has been shown. Therefore, risk quotients derived from soil exposure may underestimate the risk TCAA poses to plants. Moreover, TCE and TCA have been shown to be taken up by plants and converted to TCAA, thus leading to an additional exposure route. Mono- and di-chloroacetic acids can co-occur with TCAA in the atmosphere and soil and are more phytotoxic than TCAA. The cumulative effects of TCAA and compounds with similar toxic effects found in air and soil must be considered in subsequent terrestrial ecosystem risk assessments. PMID- 15337358 TI - Assessing wastewater toxicity to activated sludge: recent research and developments. AB - Toxicants in municipal sewage treatment plant (STP) influent wastewater may inhibit the biological activity of the activated sludge and cause treatment plant process upsets. Such process upsets may be avoided if influent wastewater is monitored for toxicity and protective actions are taken when toxicity is detected. A comprehensive review of the methods that can be used for assessing wastewater toxicity to biological treatment systems was conducted several years ago and the resultant report was published in 2000 by Water Environment Research Foundation (WERF). The WERF report also specified the criteria for influent wastewater toxicity monitoring methods and suggested research needs to be addressed. A significant amount of effort was made since the publication of the WERF report to develop new assays or devices and to improve existing ones. In this manuscript, recent research and developments in methods for assessing wastewater toxicity to activated sludge were reviewed. The literature indicates that bioluminescence- and respirometry-based methods received much attention in recent research. A comparison of the new/improved methods with the criteria described in the WERF report reveals that none of these methods has been shown to meet all the specified criteria. The present review also indicates that research efforts since 2000 have not fully taken into account the criteria for influent wastewater toxicity monitoring methods and have not addressed the research needs proposed in the WERF report. PMID- 15337359 TI - Delusion's odyssey: charting the course of Victorian forensic psychiatry. PMID- 15337360 TI - Failed agency and the insanity defence. PMID- 15337361 TI - Mental impairment, moral understanding and criminal responsibility: psychopathy and the purposes of punishment. PMID- 15337362 TI - Criminal responsibility, "fleeting" states of mental impairment, and the power of self-control. PMID- 15337363 TI - A will of one's own: consciousness, control, and character. PMID- 15337364 TI - Automatism and dissociation: disturbances of consciousness and volition from a psychological perspective. PMID- 15337365 TI - Conceptual issues in assessing responsibility for actions symptomatic of mental illness. PMID- 15337366 TI - Introduction to the reviews on neuropeptide Y. AB - Neuropeptide Y (NPY) was first reported as an abundant peptide in brain tissue in 1982. Shortly thereafter, NPY was found to be a member of a peptide family consisting of the endocrine peptides pancreatic polypeptide (PP) and peptide YY (PYY). These peptides exert most of their biological effects through five G protein coupled receptors termed Y1, Y2, Y4, Y5 and y6 that mediate either inhibition adenylate cyclase or increases in intracellular calcium. Since the discovery of NPY, a robust a body of literature has developed around the potential functions of this peptide. While initial findings identified NPY is an important contributor to the regulation of feeding, body weight and blood pressure, more recent work as revealed more subtle functions of this peptide and its potential role in affective disorders, bone formation and cravings. The accompanying twelve reviews detail important developments in our understanding of the functional role of NPY. PMID- 15337367 TI - Molecular evolution of NPY receptor subtypes. AB - The neuropeptide Y (NPY) system consists in mammals of three peptides and 4-5 G protein-coupled receptors called Y receptors that are involved in a variety of physiological functions such as appetite regulation, circadian rhythm and anxiety. Both the receptor family and the peptide family display unexpected evolutionary complexity and flexibility as shown by information from different classes of vertebrates. The vertebrate ancestor most likely had a single peptide gene and three Y receptor genes, the progenitors of the Y1, Y2 and Y5 subfamilies. The receptor genes were probably located in the same chromosomal segment. Additional gene copies arose through the chromosome quadruplication that took place before the emergence of jawed vertebrates (gnathostomes) whereupon differential losses of the gene copies ensued. The inferred ancestral gnathostome gene repertoire most likely consisted of two peptide genes, NPY and PYY, and no less than seven Y receptor genes: four Y1-like (Y1, Y4/a, Y6, and Yb), two Y2 like (Y2 and Y7), and a single Y5 gene. Whereas additional peptide genes have arisen in various lineages, the most common trend among the Y receptor genes has been further losses. Mammals have lost Yb and Y7 (the latter still exists in frogs) and Y6 is a pseudogene in several mammalian species but appears to be still functional in some. One challenge is to find out if mammals have been deprived of any functions through these gene losses. Teleost fishes like zebrafish and pufferfish, on the other hand, have lost the two major appetite stimulating receptors Y1 and Y5. Nevertheless, teleost fishes seem to respond to NPY with increased feeding why some other subtype probably mediates this effect. Another challenge is to deduce how Y2 and Y4 came to evolve an inhibitory effect on appetite. Changes in anatomical distribution of receptor expression may have played an important part in such functional switching along with changes in receptor structures and ligand preferences. PMID- 15337368 TI - Novel modified and radiolabelled neuropeptide Y analogues to study Y-receptor subtypes. AB - To better characterize neuropeptide Y (NPY) receptor subtypes, several novel selective analogues of the NPY sequence have been developed in the last few years. These analogues show high selectivity towards single NPY receptors and facilitate insights into the localisation and physiological role of each receptor subtype in health and disease. Besides the important role of NPY in the regulation of food intake and other physiological functions, recently NPY receptors have been identified to play an important role in several types of cancer. To further enlighten all these numerous functions the radioactive labelling of peptides is still an indispensable and versatile tool. By using the appropriate radioisotope and labelling strategy several different radioisotopes can be incorporated into the sequence of subtype selective NPY analogues. Depending on the radioisotope the resulting radiolabelled NPY analogues can be used for diagnostic or therapeutic applications and enable us to better investigate single receptor subtypes in vitro and in vivo. PMID- 15337369 TI - Characterization of a new neuropeptide Y Y5 agonist radioligand: [125I][cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP. AB - In order to optimally characterize a class of neuropeptide Y (NPY) receptors expressed in a tissue enriched with multiple subtypes (Y1, Y2, Y4 and Y5) and to establish its detailed distribution, it is critical to use highly selective and specific probes that possess very low non-specific binding. In that context, we recently reported on the development of [125I][hPP(1-17), Ala31, Aib32]NPY as Y5 receptor radioligand. However, the non-specific binding obtained with this radioligand was too high to allow for detailed receptor autoradiography studies [Br. J. Pharmacol. 139 (2003) 1360]. Iodinated [cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP may represent a better Y5 radioligand in that regard. Accordingly, [125I][cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP binding was investigated in rat brain membrane homogenates and its specificity and selectivity established in rat Y1, Y2, Y4 and Y5 transfected HEK293 cells. No specific binding was detected in HEK293 cells transfected with the rat Y1, Y2 or Y4 receptors, while saturable binding was observed in cells transfected with the rat Y5 receptor cDNA and in rat brain membrane homogenates (KD of 0.5-0.7 nM). Competition binding experiments performed in rat brain membrane homogenates demonstrated that specific [125I][cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP binding was competed with nanomolar affinities by Y5 agonists and antagonists such as [Leu31,Pro34]PYY, PYY(3-36), [cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP, [Ala31, Aib32]NPY, [hPP(1-17), Ala31, Aib32]NPY, CGP71683A and JCF109, but not by Y1 (BIBP3226 and BIBO3304), Y2 (BIIE0246) and Y4 (GR231118) ligands. Non-specific binding was also lower than that reported for [125I][hPP(1-17), Ala31, Aib32]NPY. Interestingly, detailed analysis of competition binding curves obtained with [Leu31, Pro34]PYY, hPP, PYY(3-36) and [cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP against specific [125I][cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP sites were best fitted to a two-site model. Additionally, receptor autoradiography studies revealed the presence of specific [125I][cPP(1-7), NPY(19 23), Ala31, Aib32, Gln34]hPP binding sites in the lateral septum and area postrema while other brain regions contained much lower levels of specific binding. Taken together, these data suggest that [125I][cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP represents a useful tool to study the unique feature of the Y5 receptor subtype. PMID- 15337370 TI - Distribution of NPY receptors in the hypothalamus. AB - Neuropeptide Y (NPY) neurons abundantly innervate the hypothalamus, where NPY is involved in the regulation and integration of a broad range of homeostatic functions. In order to understand NPY-mediated behavioral, autonomic and neuroendocrine effects, it is important to characterize in detail the distribution of the hypothalamic NPY receptors. In this review, we briefly summarize the origin of NPY and its two related peptides, peptide YY and pancreatic polypeptide in the hypothalamus. Moreover, based on the results obtained with histological techniques such as in situ hybridization, immunohistochemistry and ligand binding, we summarize data on the hypothalamic distribution of the known NPY receptors, the Y1 Y2, Y4 and Y5 receptors as best characterized to date. These NPY receptors are found with individual distribution patterns in many hypothalamic neurons including neuroendocrine motoneurons, magnocellular neurosecretory neurons and numerous neurons connecting the hypothalamus with the limbic and the autonomic nervous systems. The histochemical analyses allow characterization of coexisting molecules and in this way definition of the neurochemistry of NPY circuitries. By showing coexistence of various NPY receptors they provide a morphological basis for in vitro studies showing heterodimerization of NPY receptors. The NPY neurons and their circuitries underlie the integrative role of NPY as a pleiotropic neuropeptide in the regulation of homeostasis. PMID- 15337371 TI - NPY and Y receptors: lessons from transgenic and knockout models. AB - Neuropeptide Y (NPY) in the central nervous system is a major regulator of food consumption and energy homeostasis. It also regulates blood pressure, induces anxiolysis, enhances memory retention, affects circadian rhythms and modulates hormone release. Five Y receptors (Y1, Y2, Y4, Y5 and Y6) are known to mediate the action of NPY and its two other family members, peptide YY (PYY) and pancreatic polypeptide (PP). Increased NPY signaling due to elevated NPY expression in the hypothalamus leads to the development of obesity and its related phenotypes, Type II diabetes and cardiovascular disease. Dysregulation in NPY signaling also causes alterations in bone formation, alcohol consumption and seizure susceptibility. The large number of Y receptors has made it difficult to delineate their individual contributions to these physiological processes. However, recent studies analysing NPY and Y receptor overexpressing and knockout models have started to unravel some of the different functions of these Y receptors. Particularly, the use of conditional knockout models has made it possible to pinpoint a specific function to an individual Y receptor in a particular location. PMID- 15337372 TI - NPY and cohorts in regulating appetite, obesity and metabolic syndrome: beneficial effects of gene therapy. AB - Neuropeptide Y is the most potent physiological appetite transducer known. The NPY network is the conductor of the hypothalamic appetite regulating orchestra in the arcuate nucleus-paraventricular nucleus (ARC-PVN) of the hypothalamus. NPY and cohorts, AgrP, GABA and adrenergic transmitters, initiate appetitive drive directly through Y1, Y5, GABAA and alpha1 receptors, co-expressed in the magnocellular PVN (mPVN) and ARC neurons and by simultaneously repressing anorexigenic melanocortin signaling in the ARC-PVN axis. The circadian and ultradian rhythmicities in NPY secretion imprint the daily circadian and episodic feeding patterns. Although a number of afferent hormonal signals from the periphery can directly modulate NPYergic signaling, the reciprocal circadian and ultradian rhythmicities of anorexigenic leptin from adipocytes and orexigenic ghrelin from stomach, encode a corresponding pattern of NPY discharge for daily meal patterning. Subtle and progressive derangements produced by environmental and genetic factors in this exquisitely intricate temporal relationship between the two opposing humoral signals and the NPY network promote hyperphagia and abnormal rate of weight gain culminating in obesity and attendant metabolic disorders. Newer insights at cellular and molecular levels demonstrate that a breakdown of the integrated circuit due both to high and low abundance of NPY at target sites, underlies hyperphagia and increased adiposity. Consequently, interruption of NPYergic signaling at a single locus with NPY receptor antagonists may not be the most efficacious therapy to suppress hyperphagia and obesity. Central leptin gene therapy in rodents has been shown to subjugate, i.e. bring under homeostatic control, NPYergic signaling and suppress the age-related and dietary obesity for extended periods and thus shows promise as a newer treatment modality to curb the pandemic of obesity and metabolic syndrome. PMID- 15337373 TI - The NPY system in stress, anxiety and depression. AB - NPY antagonizes behavioral consequences of stress through actions within the brain. Behavioral anti-stress actions of NPY are noteworthy in that (1) their magnitude surpasses that of other endogenous compounds; (2) they are produced across a wide range of animal models, normally thought to reflect different aspects of emotionality. This suggests that NPY acts with a high potency on a common core mechanism of emotionality and behavioral stress responses. Behavioral studies in genetically modified animals support this hypothesis. Increased emotionality is seen upon inactivation of NPY transmission, while the opposite is found when NPY signalling is made overactive. Several brain structures are involved in mediating anti-stress actions of NPY, with the most extensive evidence available for amygdala and hippocampus, and some evidence for regions within the septum, and locus coeruleus. Antistress actions of NPY are mimicked by Y1-receptor agonists, and blocked by Y1 antagonists, although Y5 receptors may substitute for Y1 actions in some cases. Blockade of Y2 receptors produces anti stress effects indistinguishable from those produced by Y1 agonism, presumably through potentiation of presynaptic release of endogenous NPY. Together, available data point to the potential of the NPY system as a target for novel pharmacological treatments of stress-related disorders, including anxiety and depression. Development of Y2 antagonists presently appears to offer the most promising strategy for developing these clinical treatments. PMID- 15337374 TI - Interactions between NPY and CRF in the amygdala to regulate emotionality. AB - Neuropeptide Y (NPY) is one of the most abundant peptides in the central nervous system and currently there are four known receptor subtypes Y1, Y2, Y4 and Y5. Central NPY and its receptors have been implicated in a variety of physiological processes such as epilepsy, sleep, obesity, learning and memory, gastrointestinal regulation, alcoholism, depression and anxiety. The localization of these receptors within the brain is consistent with the roles mentioned, as they are found in varying density within the limbic structures, such as the hippocampal formation, amygdala, hypothalamus and septum. It is well understood that NPY produces anxiolytic responses following central administration under stressful or anxiety-provoking situations. In contrast, central administration of the neuropeptide corticotropin-releasing factor (CRF) produces anxiogenic behaviors. It has been proposed that NPY counteracts the effects of CRF to maintain no net change in emotional state, e.g., emotional homeostasis. In this article, we review the scientific literature describing the NPY-CRF relationship, specifically as it relates to the modulation of the CRF-mediated stress responses via the amygdala, a key forebrain structure involved in the regulation of emotional states. PMID- 15337375 TI - A role for neuropeptide Y in neurobiological responses to ethanol and drugs of abuse. AB - In recent years, evidence has emerged suggesting that neuropeptide Y (NPY) is involved with neurobiological responses to ethanol and other drugs of abuse. Here, we provide an overview of physiological, pharmacological, and genetic research showing that: (A) administration of ethanol, as well as ethanol withdrawal, alter central NPY expression, (B) NPY modulates ethanol consumption under certain conditions, and (C) NPY signaling modulates the sedative effects of several drugs, including ethanol, sodium pentobarbital, and ketamine. Evidence suggesting possible mechanism(s) by which NPY signaling modulates ethanol consumption are considered. It is suggested that NPY may influence ethanol consumption by regulating basal levels of anxiety, by modulating the sedative effects of ethanol, and/or by modulating ethanol's rewarding properties. PMID- 15337376 TI - Overexpression of NPY and Y2 receptors in epileptic brain tissue: an endogenous neuroprotective mechanism in temporal lobe epilepsy? AB - Recurrent epileptic seizures in the rat enhance the expression of neuropeptide Y (NPY) and its mRNA in various brain areas including the hippocampus, cerebral cortex and the amygdala. In the hippocampus, the most prominent expression of NPY is observed in mossy fibers and in GABAergic interneurons. At the same time, expression of Y2 receptors is also increased whereas Y1 receptors are reduced. Similar changes in Y1 and Y2 receptors were observed in the hippocampus of patients with temporal lobe epilepsy (TLE). In contrast to the rat, NPY expression is not enhanced in mossy fibers in TLE. In the same tissue, surviving NPY interneurons show marked axonal sprouting into areas innervated by mossy fibers (dentate hilus, stratum lucidum, inner molecular layer of the dentate gyrus). Stimulation of presynaptic Y2 receptors inhibits glutamate release, and exert an anticonvulsant action in experimental models. Y1 receptors mediate a weak excitatory component of NPY action. These findings suggest that changes in the NPY system induced by seizures represent an endogenous adaptive mechanism aimed at counteracting hyperexcitability underlying epileptic activity. This concept is strongly supported by evidence that genetically modified rats overexpressing the NPY gene are less susceptible to seizures while deletion of NPY or Y2 receptor genes results in increased susceptibility to seizures. PMID- 15337377 TI - Neuropeptide Y and seizures: effects of exogenously applied ligands. AB - The endogenous NPY system in the brain is centrally involved in seizure regulation. The present paper reviews the evidence that exogenously applied NPY receptor ligands can inhibit epileptic seizures in various rodent in vitro and in vivo models. Agonists at Y2 and/or Y5 receptors and antagonists at Y1 receptors appear to inhibit seizures, depending on the seizure model studied. Although progress has been made, further studies are needed using transgenic animals as well as novel selective agonists and antagonists to firmly identify the NPY receptors mediating antiepileptic effects. This may lead to the development of future antiepileptic drug treatments targeting the NPY system. PMID- 15337378 TI - Neuropeptide Y and epilepsy: recent progress, prospects and controversies. AB - Neuropeptide Y (NPY), a 36 amino-acid member of the pancreatic polypeptide family, has received considerable attention in recent years as an endogenous modulator of epileptic activity. Prominently expressed in brain regions involved in seizure generation and propagation, NPY can exert powerful effects on synaptic transmission. Here, we discuss the anti-epileptic actions of NPY and receptor subtypes responsible. PMID- 15337379 TI - Importance of NPY Y1 receptor-mediated pathways: assessment using NPY Y1 receptor knockouts. AB - The peptidic neurotransmitter neuropeptide Y (NPY) has been functionally implicated in feeding behavior, cardiovascular regulation, control of neuroendocrine axes, affective disorders, seizures, and memory retention. At least five different receptors mediate NPY actions. In particular, the Y1 receptor appears to be involved in a variety of NPY-induced pathways. This review summarizes the main findings resulting from the use of mice lacking NPY Y1 receptor expression. Interestingly, the overall phenotype of Y1 knockouts mimics metabolic syndrome, which is characterized by obesity, a prediabetic state, and a susceptibility to develop hypertension. PMID- 15337380 TI - The World Bank is finally embracing science. PMID- 15337381 TI - Paediatric research should take centre stage. PMID- 15337382 TI - Oseltamivir-resistant influenza? PMID- 15337383 TI - Reconstruction of human mandible by tissue engineering. PMID- 15337384 TI - Extending the use of the glycaemic index: beyond diabetes? PMID- 15337385 TI - Asking parents unaskable questions. PMID- 15337386 TI - Appropriate use of opioids for persistent non-cancer pain. PMID- 15337387 TI - Kaposi's sarcoma. PMID- 15337388 TI - Faecal occult-blood screening in Burgundy. PMID- 15337389 TI - Mobilising support to sustain political will is the key to progress in reproductive health. PMID- 15337390 TI - As we said... PMID- 15337391 TI - Wounded by words. PMID- 15337392 TI - Observational versus randomised trial evidence. PMID- 15337394 TI - Observational versus randomised trial evidence. PMID- 15337395 TI - Observational versus randomised trial evidence. PMID- 15337397 TI - Observational versus randomised trial evidence. PMID- 15337398 TI - HIV/HCV coinfection, HAART, and liver-related mortality. PMID- 15337400 TI - The right answer. PMID- 15337401 TI - Resistant influenza A viruses in children treated with oseltamivir: descriptive study. AB - BACKGROUND: Oseltamivir is an effective inhibitor of influenza virus neuraminidase. Although viruses resistant to oseltamivir emerge less frequently than those resistant to amantadine or rimantadine, information on oseltamivir resistant viruses arising during clinical use of the drug in children is limited. Our aim was to investigate oseltamivir resistance in a group of children treated for influenza. METHODS: We analysed influenza A viruses (H3N2) collected from 50 children before and during treatment with oseltamivir. We sequenced the genes for neuraminidase and haemagglutinin and studied the mutant neuraminidases for their sensitivity to oseltamivir carboxylate. FINDINGS: We found neuraminidase mutations in viruses from nine patients (18%), six of whom had mutations at position 292 (Arg292Lys) and two at position 119 (Glu119Val), which are known to confer resistance to neuraminidase inhibitors. We also identified another mutation (Asn294Ser) in one patient. Sensitivity testing to oseltamivir carboxylate revealed that the neuraminidases of viruses that have an Arg292Lys, Glu119Val, or Asn294Ser mutation were about 10(4)-10(5)-fold, 500-fold, or 300 fold more resistant than their pretreatment neuraminidases, respectively. Oseltamivir-resistant viruses were first detected at day 4 of treatment and on each successive day of the study. More than 10(3) infectious units per mL of virus were detected in some of the patients who did not shed drug-resistant viruses, even after 5 days of treatment. INTERPRETATION: Oseltamivir-resistant mutants in children being treated for influenza with oseltamivir arise more frequently than previously reported. Furthermore, children can be a source of viral transmission, even after 5 days of treatment with oseltamivir. PMID- 15337402 TI - Growth and transplantation of a custom vascularised bone graft in a man. AB - BACKGROUND: A major goal of research in bone transplantation is the ability to avoid creation of secondary bone defects. We aimed to repair an extended mandibular discontinuity defect by growth of a custom bone transplant inside the latissimus dorsi muscle of an adult male patient. METHODS: Three-dimensional computed tomography (CT) scanning and computer-aided design techniques were used to produce an ideal virtual replacement for the mandibular defect. These data were used to create a titanium mesh cage that was filled with bone mineral blocks and infiltrated with 7 mg recombinant human bone morphogenetic protein 7 and 20 mL of the patient's bone marrow. Thus prepared, the transplant was implanted into the latissimus dorsi muscle and 7 weeks later transplanted as a free bone-muscle flap to repair the mandibular defect. FINDINGS: In-vivo skeletal scintigraphy showed bone remodelling and mineralisation inside the mandibular transplant both before and after transplantation. CT provided radiological evidence of new bone formation. Postoperatively, the patient had an improved degree of mastication and was satisfied with the aesthetic outcome of the procedure. INTERPRETATION: Heterotopic bone induction to form a mandibular replacement inside the latissimus dorsi muscle in a human being is possible. This technique allows for a lower operative burden compared with conventional techniques by avoiding creation of a secondary bone defect. It also provides a good three-dimensional outcome. PMID- 15337403 TI - Mortality and incidence of cancer during 10-year follow-up of the Scandinavian Simvastatin Survival Study (4S). AB - BACKGROUND: The effects of cholesterol-lowering treatment with statins on mortality and risk of cancer beyond the usual 5-6-year trial periods are unknown. We extended post-trial follow-up of participants in the Scandinavian Simvastatin Survival Study (4S) to investigate cause-specific mortality and incidence of cancer 5 years after closure of the trial. METHODS: 4S was a randomised double blind trial of simvastatin or placebo in patients with coronary heart disease, serum total cholesterol 5.5-8.0 mmol/L, and serum triglycerides 2.5 mmol/L or lower. The double-blind period lasted for a median of 5.4 years (range for survivors 4.9-6.3) and ended in 1994. After the trial, most patients in both groups received open-label lipid-lowering treatment. National registers were used to assess mortality and causes of death and cancer incidence in the original treatment groups for a median total follow-up time of 10.4 years (range for survivors 9.9-11.3). Analysis was by intention to treat. FINDINGS: 414 patients originally allocated simvastatin and 468 assigned placebo died during the 10.4 year follow-up (relative risk 0.85 [95% CI 0.74-0.97], p=0.02), a difference largely attributable to lower coronary mortality in the simvastatin group (238 vs 300 deaths; 0.76 [0.64-0.90], p=0.0018). 85 cancer deaths arose in the simvastatin group versus 100 in the placebo group (0.81 [0.60-1.08], p=0.14), and 227 incident cancers were reported in the simvastin group versus 248 in the placebo group (0.88 [0.73-1.05], p=0.15). Incidence of any specific type of cancer did not rise in the simvastatin group. INTERPRETATION: Simvastatin treatment for 5 years in a placebo-controlled trial, followed by open-label statin therapy, was associated with survival benefit over 10 years of follow-up compared with open-label statin therapy for the past 5 years only. No difference was noted in mortality from and incidence of cancer between the original simvastatin group and placebo group. PMID- 15337404 TI - Effects of dietary glycaemic index on adiposity, glucose homoeostasis, and plasma lipids in animals. AB - BACKGROUND: Clinical studies suggest a role for dietary glycaemic index (GI) in bodyweight regulation and diabetes risk. However, partly because manipulation of GI can produce changes in potentially confounding dietary factors such as fibre content, palatability, and energy density, its relevance to human health remains controversial. This study examined the independent effects of GI in animals. METHODS: Partially pancreatectomised male Sprague-Dawley rats were given diets with identical nutrients, except for the type of starch: high-GI (n=11) or low-GI (n=10). The animals were fed in a controlled way to maintain the same mean bodyweight in the two groups for 18 weeks. Further experiments examined the effects of GI in rats in a cross-over design and C57BL/6J mice in a parallel design. FINDINGS: Despite having similar mean bodyweight (547.9 [SE 13.4] vs 549.2 [15.2] g), rats given high-GI food had more body fat (97.8 [13.6] vs 57.3 [7.2] g; p=0.0152) and less lean body mass (450.1 [9.6] vs 491.9 [11.7] g; p=0.0120) than those given low-GI food. The high-GI group also had greater increases over time in the areas under the curve for blood glucose and plasma insulin after oral glucose, lower plasma adiponectin concentrations, higher plasma triglyceride concentrations, and severe disruption of islet-cell architecture. Mice on the high-GI diet had almost twice the body fat of those on the low-GI diet after 9 weeks. INTERPRETATION: These findings provide a mechanistic basis for interpretation of studies of GI in human beings. RELEVANCE TO PRACTICE: The term GI describes how a food, meal, or diet affects blood sugar during the postprandial period. GI as an independent factor can cause obesity and increase risks of diabetes and heart disease in animals. Use of low-GI diets in prevention and treatment of human disease merits thorough examination. PMID- 15337405 TI - Neuronal lipofuscinosis. PMID- 15337406 TI - A population-based nationwide study of parents' perceptions of a questionnaire on their child's death due to cancer. AB - A proposed nationwide postal questionnaire to Swedish parents who had lost a child due to cancer between 1992 and 1997 was denied approval by the local ethics committee. However, a pilot study to assess the harm and benefit of the questionnaire was approved. 95% of parents found the pilot study valuable; thus, we were allowed to proceed with the main study, which consisted of 129 questions about the child's care and death and five about the parents' perceptions of the study. 423 (99%) parents found the investigation valuable, 285 (68%) were positively affected, and 123 (28%) were negatively affected (10 [2%] of whom, very much). Although the numerical data cannot be directly translated to ethical conclusions, they can provide guidance for future ethical decisions. PMID- 15337407 TI - Efficacy of 10-day melarsoprol schedule 2 years after treatment for late-stage gambiense sleeping sickness. AB - In 2000, we reported that a new short treatment schedule of melarsoprol was not worse than the longer and demanding standard treatment for late-stage human African trypanosomiasis. This alternative schedule was assessed in an open, randomised clinical equivalence trial of 500 patients in Angola. 24 h after treatment, all patients were parasite free. Of 442 patients, 12 (3%) had relapsed after 1 year, of whom seven (3%) had had standard treatment and five (2%) the alternative treatment. After 2 years, 23 (5%) relapsing patients were reported, 11 (5%) in the standard treatment group and 12 (6%) in the new group. The results at the 2-year follow-up support and strengthen our previous findings. PMID- 15337408 TI - Treatment of stable chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a readily diagnosable disorder that responds to treatment. Smoking cessation can reduce symptoms and prevent progression of disease. Bronchodilator therapy is key in improvement of lung function. Three classes of bronchodilators-beta agonists, anticholinergics, and theophylline-are available and can be used individually or in combination. Inhaled glucocorticoids can also improve airflow and can be combined with bronchodilators. Inhaled glucocorticoids, in addition, might reduce exacerbation frequency and severity as might some bronchodilators. Effective use of pharmacotherapy in COPD needs integration with a rehabilitation programme and successful treatment of co-morbidities, including depression and anxiety. Treatment for stable COPD can improve the function and quality of life of many patients, could reduce admissions to hospital, and has been suggested to improve survival. PMID- 15337409 TI - Clinical trials in children. AB - The imperative to undertake randomised trials in children arises from extraordinary advances in basic biomedical sciences, needing a matching commitment to translational research if child health is to reap the benefits from this new knowledge. Unfortunately, many prescribed treatments for children have not been adequately tested in children, sometimes resulting in harmful treatments being given and beneficial treatments being withheld. Government, industry, funding agencies, and clinicians are responsible for research priorities being adult-focused because of the greater burden of disease in adults, coupled with financial and marketing considerations. This bias has meant that the equal rights of children to participate in trials has not always been recognised. This is changing, however, as the need for clinical trials in children has been increasingly recognised by the scientific community and broader public, leading to new legislation in some countries making trials of interventions mandatory in children as well as adults before drug approval is given. Trials in children are more challenging than those in adults. The pool of eligible children entering trials is often small because many conditions are uncommon in children, and the threshold for gaining consent is often higher and more complex because parents have to make decisions about trial participation on behalf of their child. Uncertain about what is best, despite supporting the notion of trials in principle, parents and paediatricians generally opt for the new intervention or for standard care rather than trial participation. In this review, we explore issues relating to trial participation for children and suggest some strategies for improving the conduct of clinical trials involving children. PMID- 15337410 TI - Improving surgical consent. PMID- 15337411 TI - A Kendo player with haemoptysis. PMID- 15337412 TI - Can low-magnification stereomicroscopy reveal diet? AB - A new method of scoring dental microscopic use wear, initially developed for and applied to extant and extinct ungulates, is here applied to primates, and the efficacy of the method as a tool for diagnosing diet in both ungulates and primates is established. The method employs standard refractive light microscopy instead of scanning electron microscopy (SEM), and all use-wear features are counted or scored under low magnification (35 x). We use measurement systems analysis (variance components analysis of sources of measurement error) to evaluate the consistency and reproducibility of measurements using this method. The method is shown to have low intra- and inter-observer measurement error, and to effectively distinguish among graminivores, folivores, and frugivores. It can also be used to identify seed predators and to diagnose hard-object feeding. The method is also shown to be robust to the selection of measurement site; it works equally well when applied to upper or to lower molars. Finally, we use analysis of variance to examine the consistency of the signals across mammalian orders, and discriminant function analysis to develop dietary diagnostic tools for a set of "classified" primates with known diets. We test the success of these tools not merely by examining their a posteriori classification "success," but by using them to construct predicted dietary profiles for a sample of unclassified extant primate species, again with known diets. PMID- 15337413 TI - Dental use wear in extinct lemurs: evidence of diet and niche differentiation. AB - A new technique for molar use-wear analysis is applied to samples of all 16 species of extinct lemurs with known dentitions, as well as to a large comparative sample of extant primates. This technique, which relies on the light refractive properties of wear pits and scratches as seen under a standard stereoscopic microscope, has shown itself to be effective in distinguishing the diets of ungulates and extant primates. We draw dietary inferences for each of the 16 extinct lemur species in our database. There is a strong phylogenetic signal, with the Palaeopropithecidae showing use-wear signatures similar to those of the Indriidae; extinct lemurids (Pachylemur spp.) showing striking similarities to extant lemurids (except Hapalemur spp.); and Megaladapis showing similarities to Lepilemur spp. Only the Archaeolemuridae have dietary signatures unlike those of any extant lemurs, with the partial exception of Daubentonia. We conclude that the Archaeolemuridae were hard-object feeders; the Palaeopropithecidae were seed predators, consuming a mixed diet of foliage and fruit to varying degrees; Pachylemur was a fruit-dominated mixed feeder, but not a seed predator; and all Megaladapis were leaf browsers. There is no molar use wear evidence that any of the extinct lemurs relied on terrestrial foods (C4 grasses, tubers, rhizomes). This has possible implications for the role of the disappearance of wooded habitats in the extinction of lemurs. PMID- 15337414 TI - New hominin first metatarsal (SK 1813) from Swartkrans. AB - A recently recognized hominin hallucal metatarsal, SK 1813, from Swartkrans bears a suite of primitive and derived traits. Comparisons with extant apes, modern humans, SKX 5017, and Stw 562 reveals similar morphology in all three fossils and that these early hominins, while bipedal, possessed a unique toe-off mechanism. The implications of this are that both primitive and derived traits must be used to establish the total biomechanical pattern. PMID- 15337415 TI - A morphometric analysis of maxillary molar crowns of Middle-Late Pleistocene hominins. AB - This study explores the significance of shape differences in the maxillary first molar crowns of Neandertals and anatomically modern humans. It uses morphometric analysis to quantify these differences and to investigate how the orientation of major cusps, relative cusp base areas and occlusal polygon area influence crown shape. The aims of this study were to 1) quantify these data to test whether the tooth shapes of Neandertals and anatomically modern humans differ significantly and 2) to explore if either of the shapes is derived relative to earlier fossil hominins. Data were collected from digital occlusal photographs using image processing software. Cusp angles, relative cusp base areas and occlusal polygon areas were measured on Neandertals (n=15), contemporary modern humans (n=62), Upper Paleolithic humans (n=6), early anatomically modern humans (n=3) and Homo erectus (n=3). Univariate and multivariate statistical tests were used to evaluate the differences between contemporary modern humans and Neandertals, while the much sparser data sets from the other fossil samples were included primarily for comparison. Statistically significant differences reflecting overall crown shape and internal placement of the crown apices were found. Neandertals are distinguished from contemporary humans by possessing maxillary first molars that 1) are markedly skewed; 2) possess a narrower distal segment of the occlusal polygon compared to the mesial segment; 3) possess a significantly smaller metacone and a significantly larger hypocone; and 4) possess a significantly smaller relative occlusal polygon area reflecting internally placed cusps. Differences in relative cusp base areas of the hypocone and metacone may contribute to the shape differences observed in Neandertals. However, early anatomically modern humans possessing a pattern of relative cusp base areas similar to Neandertals lack their unusual shape. That the morphology observed in non-Neandertal fossil hominins is more anatomically modern human-like than Neandertal-like, suggests that this distinctive morphology may be derived in Neandertals. PMID- 15337416 TI - The randomized trials of breast cancer screening: what have we learned? AB - Eight randomized controlled trials of mammography screening have been conducted to date. In addition to evaluating the efficacy of screening with an experimental design, the trials provided investigators with access to information about breast cancers much earlier in their development than had previously been available. The trials of mammographic screening provide conclusive evidence that the policy of offering screening is associated with a significant and substantial reduction in breast cancer mortality. PMID- 15337417 TI - Adverse effects of screening mammography. AB - The main risks and other adverse consequences from screening mammography include discomfort from breast compression, patient recall for additional imaging, and false positive biopsies. Although these risks affect a larger number of women than those who benefit from screening, the risks are less consequential than the life-sparing benefits from early detection. Radiation risk, even for multiple screenings, is negligible at current mammography doses. Anxiety before screening or resulting from supplementary imaging work-up, short-term follow-up, cyst aspiration, and biopsy has not dampened the enthusiasm of most women for the value of early detection. PMID- 15337418 TI - Pathology of high-risk breast lesions and ductal carcinoma in situ. AB - This article illustrates and provides an update on lesions considered to be precursors of mammary carcinoma or indicators of elevated risk for subsequent development of carcinoma.A review of usual ductal hyperplasia and ductal carcinoma in situ is given to provide a background reference for comparison with the high-risk lesions. Lesions illustrated and described as high-risk include atypical ductal hyperplasia, atypical columnar cell hyperplasia, lobular carcinoma in situ, atypical lobular hyperplasia, and proliferative radial scar lesions. PMID- 15337419 TI - Screening mammography practice essentials. AB - A successful screening mammography practice has three directives. The first directive is quality mammography interpretation, which results in detection of a high percentage of early stage breast cancers, an acceptable recall rate, and an acceptable biopsy rate and yield. The second directive is providing a cost efficient service. The third directive is access for as many eligible women as possible. Strategies that have helped improve screening mammography access for underserved women are discussed in this article. PMID- 15337420 TI - Supplemental screening sonography in dense breasts. AB - In single-center trials across 42,838 examinations, 150 (0.35%) cancers were identified only sonographically in average-risk women. Over 90% of the 126 women with sonographically depicted cancers had dense or heterogeneously dense parenchyma. Of the 150 cancers, 141 (94%) were invasive, with a mean size of 9 to 11 mm across the series. Over 90% were node-negative. A3-year multicenter trial of screening sonography in high-risk women, blinded to the results of mammography, opened for enrollment April 2004,funded by the Avon Foundation and National Cancer Institute through the American College of Radiology Imaging Network (ACRIN Protocol 6666). If the trial is successful,the results will provide a rational basis for supplemental screening sonography in women with dense breasts. PMID- 15337421 TI - Diagnostic mammography. AB - When evaluating mammograms, one looks for masses, areas of asymmetry or architectural distortion, and microcalcifications. This article discusses the imaging evaluation and the management of lesions found on screening and diagnostic mammography, with the focus on commonly encountered questions and dilemmas. PMID- 15337422 TI - Digital mammography. AB - Despite its technical advantages, early clinical trials comparing digital mammography with film mammography for screening have been somewhat disappointing. Digital mammography,however, is in its infancy and can be expected to improve more rapidly than film mammography. Some areas of improvement being observed now include the development of new detector technologies; more powerful and better designed interpretation workstations; and novel advanced applications, such as tomosynthesis and contrast-enhanced mammography, which are not possible with standard film mammography. PMID- 15337423 TI - Molecular imaging in breast cancer. AB - Since the 1960s, bone scanning has played a major role in the management of breast cancer. In the last decade, however, the role of radionuclide molecular imaging has expanded significantly in the clinical management of breast cancer because of fluorodeoxyglucose positron emission tomography, mammoscintigraphy, and sentinel lymph node techniques. Molecular imaging also is instrumental in drug development,gene therapy, and in basic science research of breast cancer. This article provides a comprehensive review of the role of molecular imaging of breast cancer in clinical practice and reports on the current state of research in this field. PMID- 15337424 TI - Problem-solving ultrasound. AB - This article addresses the accepted indications for using ultrasound alone or in conjunction with other imaging techniques in solving problems of breast diagnosis and guiding patient management. The indications include evaluation of palpable and mammographic masses; guidance of interventional procedures; evaluating problems with implants; radiation treatment planning; and as the initial imaging technique in young,pregnant, and lactating patients. Evolving indications and controversial applications for breast ultrasound are also discussed. PMID- 15337425 TI - Problem solving MR imaging of the breast. AB - MR imaging is emerging as a valuable adjunct to mammography and ultrasound for the evaluation of the breast. Breast MR imaging has high sensitivity for the detection of breast cancer but suffers from a relatively low specificity. There is growing experience with this imaging modality as applied to breast diseases and more standardization in obtaining and interpreting these examinations is occurring, including the introduction ofa reporting lexicon similar to that for mammography. Breast MR imaging can be used for problem solving in determination of the local extent of disease in women with newly diagnosed breast cancer and for the evaluation of cases that remain inconclusive despite thorough mammographic and sonographic evaluation. PMID- 15337426 TI - Image-guided breast biopsy and management of high-risk lesions. AB - Across several series, the sensitivity of sonographically guided 14-gauge core biopsy is 95%, and the repeat biopsy rate averages 11%. Success of stereotactic biopsy requires obtaining larger volumes of tissue when sampling calcifications; use of vacuum-assisted devices results in comparable sensitivities. For MR imaging-guided percutaneous biopsy,success rates of 95% to 99% have been achieved. Independent of guidance method or the amounts of tissue acquired, the following diagnoses on percutaneous biopsy should generally prompt excision: atypical ductal hyperplasia, lobular neoplasia, radial sclerosing lesions, benign and atypical papillary lesions, and possible phyllodes tumor. Mucocele-like lesions may merit excision. Columnar alteration without atypia probably does not require excision, although further study is needed. PMID- 15337427 TI - MR imaging--guided breast ablative therapy. AB - The integration of imaging and thermal therapy can provide a minimally invasive or even noninvasive alternative to breast surgery for small tumors. Ongoing trials seek to show safety and efficacy for laser, radiofrequency, microwave, cryoablation, and focused ultrasound surgery. To be successful, these therapies must achieve equivalent or even greater efficacy as surgical outcomes and must demonstrate total ablation of the dominant lesion with negative margins, while sparing normal tissue beyond the target tissue. Procedures have been validated by histopathology subsequent to resection. PMID- 15337428 TI - Breast imaging, standard of care, and the expert. AB - Many radiologists have the opportunity to serve as experts in determining whether a colleague's actions are within standard of care. This is an important responsibility because, in large part, the fairness and success of the judicial system determining whether medical negligence has occurred is dependent on the opinion of the medical expert. Especially in the field of breast imaging, the physician acting as an expert often has to base opinions on judgments rather than incontestable facts. To make judgments of the highest quality, the expert must possess a clear understanding of the concept of standard of care. This article focuses on the role of experts in malpractice cases dealing with mammography and the delay of diagnosis of breast cancer. PMID- 15337429 TI - Mammography: will adequate manpower exist? AB - Data exist that clearly demonstrate the dominant role of screening mammography in breast cancer mortality reduction with an ever-increasing population of women who are eligible to receive the benefits of screening. The critical question now concerns the current and future availability of those who perform and interpret these life-saving examinations. PMID- 15337431 TI - 1-Phenyl-3-toluyl-4-[ortho-1'-(N-ethyl-2'-methylpropylamine)]phenylpyrazole, synthesis and evaluation of the in vitro antifungal activity against Botrytis cinerea and Fusarium oxysporum. AB - A novel antifungal pyrazole derivative was synthesized. Designated 1-phenyl-3 toluyl-4-[ortho-1'-(N-ethyl-2'-methylpropylamine)]phenylpyrazole, the compound exerted an antifungal effect toward Botrytis cinerea and Fusarium oxysporum. In fact, our results clearly show that mycelial growth and conidial germination of both fungi were blocked by the compound. Indeed, a 96-well microbioassay procedure was used for fast and easy evaluation of minimal inhibitory concentration (MIC). The MIC values for B. cinerea and F. oxysporum were 25 and 36 microg/ml, respectively. PMID- 15337432 TI - Acetylsalicylic acid as a potential regulator of prolidase-convertible pro-drugs in control and neoplastic cells. AB - Proline analogue of melphalan (Mel-pro) is one of the pro-drugs activated by prolidase, cytoplasmic imidodipeptidase highly expressed in some neoplastic tissues. In order to limit the action of prolidase on the pro-drug in normal cells, prolidase inhibitor, acetylsalicylic acid (ASA), was tested in fibroblasts (showing average prolidase activity for normal cells) and in MDA-MB 231 breast cancer cells (showing elevated activity of the enzyme). The effect of Mel-pro in the presence and absence of ASA on prolidase activity (colorimetric assay), DNA biosynthesis (3H-thymidine incorporation assay), cytotoxicity (tetrazoline assay) and ability to penetrate cell membrane (thin layer chromatography) in both type of cells was measured. It has been found that 5 mM ASA significantly decreased conversion of Mel-pro to Mel in cultured fibroblasts as well as it decreased cytotoxicity and the effect of this drug on DNA synthesis. In contrast, 5 mM ASA had relatively lower effect on the conversion of Mel-pro into Mel in MDA-MB 231 cells as well it had little effect on Mel-pro-induced inhibition of DNA synthesis and cell death. It suggests that ASA may serve as an inhibitor of prolidase convertible pro-drugs in normal cells. PMID- 15337433 TI - Synthesis and antibacterial evaluation of oxazolidin-2-ones structurally related to linezolid. AB - Compounds structurally related to the known antimicrobial drug linezolid were selected in order to evaluate the influence of electron-withdrawing properties and altered geometric features as a result of the N-substituent modification. After a preliminary study of molecular modeling, cinnamoyl-, pyridin- and pyrimidinoxazolidin-2-ones were synthesized. None of the new compounds showed antibacterial activity. PMID- 15337434 TI - Synthesis and antimicrobial activity of new 7 beta (benzo[a]dihydrocarbazolyloxyacetyl)-substituted cephalosporins. AB - Selected 7beta-(benzo[a]dihydrocarbazolyloxyacetyl)-substituted cephalosporins (1a-e) were synthesised and tested for their antimicrobial activity against Gram positive and Gram-negative clinical pathogens. All compounds synthesised (1a-e) showed an in vitro antimicrobial activity similar to that of ceftriaxone and cefpirome against the Gram-positive bacteria, and superior to that of penicillin and cefaclor against pen-R Staphylococcus aureus species. Like all beta-lactam agents, compounds 1a-e were in an inactive Minimum Inhibitory Concentration (MIC > 32 microg/ml) against methicillin-resistant S. aureus species. Furthermore, as expected, no cross-resistance was observed against the erythromycin-resistant Staphylococcus pyogenes strain. Finally, it is worth underlining that compounds 1a and 1e showed a similar activity to that of ceftriaxone and superior to cefaclor against penicillin-resistant Streptococcus pneumoniae isolates, a key respiratory tract infection (RTI) causing pathogen difficult to treat with currently marketed antibiotics. PMID- 15337435 TI - Effect of cytochrome P450 inhibitors (diethyl dithiocarbamate, ketoconazole and grapefruit juice) on the pharmacokinetics of all-trans-retinoic acid. AB - Diethyl dithiocarbamate (DEDTC) has been reported to be a more powerful inhibitor of all-trans-retinoic acid (ATRA) in vitro metabolism than the well-established cytochrome P450 (CYP) inhibitor ketoconazole (KC). In recent years grapefruit juice (GJ) has been shown to be able to increase the oral bioavailability of several drugs by inhibiting intestinal CYP. This study investigated the in vivo effect of these CYP inhibitors on the pharmacokinetics of ATRA. The latter was administered to rats as a constant-rate intravenous (i.v.) infusion (0.48 mg h( 1) kg(-1)) during 10 h and orally (1.6 mg kg(-1)). DEDTC (320 mg kg(-1) x 2 i.v., 6.4 and 32 mg kg(-1) per os (p.o.)) did not change the ATRA concentration-time profiles, whereas KC (320 and 32 mg kg(-1) p.o.)--with i.v. infused or orally dosed ATRA--increased the mean concentration-time curve value by 160% and 78%, respectively. A high dose of DEDTC (320 mg kg(-1) p.o.) caused a marked decrease in plasma levels of ATRA. GJ (6.4 ml kg(-1) p.o.) did not affect the plasma levels of ATRA. It is concluded that the in vivo effect of CYP inhibitors (DEDTC and KC) on the elimination rate of ATRA is qualitatively different from that expected from in vitro studies. PMID- 15337436 TI - Determination of certain drugs in binary mixtures formulations by second derivative ratio spectrophotometry and LC. AB - Spectrophotometric and high-performance liquid chromatographic (HPLC) methods are developed for simultaneous determination of three binary mixtures with overlapping spectra. The spectrophotometric method is based on the use of second derivative of the ratio spectra (2DD) for resolution of three binary mixtures of indapamide with captopril (mixture 1), cinnarizine with heptaminol acefylline (mixture 2) and amoxycillin trihydrate with flucloxacillin sodium (mixture 3). The HPLC method depends on the separation of components of binary mixtures using ODS column with mobile phase consisting of acetonitrile and 5 mM aqueous heptane sulphonic acid sodium salt in ratios of (60:40, v/v, pH 5.5) for mixture 1, (50:50, v/v, pH 3.0) for mixture 2 and (35:65, v/v, pH 4.2) for mixture 3. The proposed methods are accurate, non-destructive and successfully applied for the determination of the three binary combinations in synthetic mixtures and commercial pharmaceutical products. PMID- 15337437 TI - Spectrophotometric and LC determination of two binary mixtures containing antihistamins. AB - Several methods are developed for the determination of two binary mixtures containing cyclizine hydrochloride with pyridoxine HCl (mixture (mix.) 1); and cinnarizine with piracetam (mix. 2). The resolution of the two binary mixtures has been accomplished by using numerical spectrophotometric methods as partial least squares (PLS-1) and principal component regression applied to UV spectra of the mixture and graphical spectrophotometric method as second derivative of the ratio spectra (2DD). In addition, HPLC methods were developed depending on using RP18 column with mobile phase consisting of acetonitrile/0.05 M KH2PO4 (50:50, v/v, pH 4.0) with UV detection at 239 nm for mix. 1, and mobile phase consisting of acetonitrile/0.05 M KH2PO4/triethylamine (50:50:0.2, v/v/v, pH 3.0) with UV detection at 227 nm for mix. 2. The proposed methods were successfully applied for the determination of the two binary combinations in synthetic mixtures and commercial tablets. PMID- 15337438 TI - Comparison of two derivative spectrophotometric methods for the determination of alpha-tocopherol in pharmaceutical preparations. AB - Simple, sensitive and reliable derivative spectrophotometric methods were developed and validated for determination of alpha-tocopherol in pharmaceutical preparations. The solutions of standard and the sample were prepared in absolute ethanol. The quantitative determination of the drug was carried out using the first derivative values measured at 284, 304 nm and the second derivative values measured at 288, 296 nm. Calibration graphs constructed at their wavelengths of determination were linear in the concentration range of alpha-tocopherol using peak to zero 10-250 microg ml(-1) for first and second derivative spectrophotometric methods. Developed spectrophotometric methods in this study are accurate, sensitive, precise, reproducible, and can be directly and easily applied to Evon dragee form as pharmaceutical preparation. Statistical analysis (Student's t-test) of the obtained results showed no significant difference between the proposed two methods. PMID- 15337439 TI - The ICH guidance in practice: stress degradation studies on indinavir sulphate and development of a validated specific stability-indicating HPTLC assay method. AB - A sensitive, selective, precise and stability-indicating high-performance thin layer chromatography (HPTLC) method for analysis of indinavir sulphate both as a bulk drug and in formulations was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of carbon tetrachloride/chloroform/methanol/10% v/v ammonia (4:4.5:1.5:0.05, v/v/v/v). Densitometric analysis of indinavir sulphate was carried out in the absorbance mode at 260 nm. This system was found to give compact spots for indinavir sulphate (Rf value of 0.43 +/- 0.02, for six replicates). Indinavir sulphate was subjected to acid and alkali hydrolysis, oxidation, dry and wet heat treatment, and photo degradation. The drug undergoes degradation under acidic and basic conditions, oxidation, dry and wet heat treatment, and photo degradation. Also the degraded products were well resolved from the pure drug with significantly different Rf values. The method was validated for linearity, precision, robustness, limit of detection (LOD), limit of quantitation (LOQ), specificity and accuracy. Linearity was found to be in the range of 100-6000 ng/spot with significantly high value of correlation coefficient r2 = 0.997 +/- 0.64. The linear regression analysis data for the calibration plots showed good linear relationship with r2 = 0.999 +/- 0.002 in the working concentration range of 1000-6000 ng/spot. The LOD and LOQ were 40 and 120 ng/spot, respectively. Statistical analysis proves that the method is repeatable and specific for the estimation of the said drug. As the method could effectively separate the drug from its degradation products, it can be employed as a stability-indicating one. Moreover, the proposed HPTLC method was utilized to investigate the kinetics of acid degradation process. Arrhenius plot was constructed and activation energy was calculated. PMID- 15337440 TI - Simple high-performance liquid chromatographic determination of buspirone in human plasma. AB - A simple, rapid and sensitive high-performance liquid chromatographic method for the determination of buspirone in plasma was developed. Buspirone was isolated from plasma using protein precipitation by acetonitrile and the recovery was complete. Citalopram was used as internal standard. The chromatographic conditions were as follows: analytical 125 x 4 mm, i.d. Nucleosil C18 column (5 microm particle size), mobile phase consisting of sodium dihydrogen phosphate buffer/acetonitrile (60:40, v/v) adjusted to pH 5.5 at a flow rate of 1.0 ml min( 1), UV detection at 235 nm. The quantification limit for buspirone in plasma was 0.5 ng ml(-1). The calibration curve was linear over the concentration range 0.5 10 ng ml(-1). The inter- and intra-day assay coefficients of variation were found to be less than 8%. The present validated method was successfully used for bioequivalence studies of buspirone in human subjects. PMID- 15337441 TI - Enantiomeric separation of cizolirtine and metabolites on amylose tris (3,5 dimethylphenyl carbamate) chiral stationary phase. AB - The enantiomeric resolution of (+/-)-cizolirtine, (+/-)-cizolirtine-N-oxide, (+/ )-N-desmethylcizolirtine and (+/-)-5(alpha-hydroxybenzyl)-1-methylpyrazole was achieved on amylose tris (3,5-dimethylphenyl carbamate) chiral stationary phase known as Chiralpak AD using hexane/2-propanol/triethylamine (80:20:0.05, v/v/v) as the mobile phase. The flow rate of the mobile phase used was 1.0 ml/min with UV detection at 230 nm. The values of R(s) of the resolved enantiomers of (+/-) cizolirtine, (+/-)-cizolirtine-N-oxide, (+/-)-N-desmethylcizolirtine and (+/-) 5(alpha-hydroxybenzyl)-1-methyl pyrazole were 1.20, 0.60, 1.16 and 1.15, respectively. PMID- 15337442 TI - Particle interaction of lubricated or unlubricated binary mixtures according to their particle size and densification mechanism. AB - The aim of this study is to assess an experimental approach for technological development of a direct compression formulation. A simple formula was considered composed by an active ingredient, a diluent and a lubricant. The active ingredient and diluent were selected as an example according to their typical densification mechanism: the nitrofurantoine, a fragmenting material, and the cellulose microcrystalline (Vivapur), which is a typical visco-elastic material, equally displaying good bind and disintegrant properties. For each ingredient, samples of different particle size distribution were selected. Initially, tabletability of pure materials was studied by a rotary press without magnesium stearate. Vivapur tabletability decreases with increase in particle size. The addition of magnesium stearate as lubricant decreases tabletability of Vivapur of greater particle size, while it kept unmodified that of Vivapur of lower particle size. Differences in tabletability can be related to differences in particle particle interactions; for Vivapur of higher particle size (Vivapur 200, 102 and 101), the lower surface area develops lower surface available for bonds, while for Vivapur of lower particle size (99 and 105) the greater surface area allows high particle proximity favouring particle cohesivity. Nitrofurantoine shows great differences in compression behaviour according to its particle size distribution. Large crystals show poorer tabletability than fine crystals, further decreased by lubricant addition. The large crystals poor tabletability is due to their poor compactibility, in spite of high compressibility and plastic intrinsic deformability; in fact, in spite of the high densification tendency, the nature of the involved bonds is very weak. Nitrofurantoine samples were then mixed with Vivapurs in different proportions. Compression behaviour of binary mixes (tabletability and compressibility) was then evaluated according to diluents proportion in the mixes. The mix of either nitrofurantoine large crystals or fine crystals with cellulose microcrystalline showed a negative interaction in all proportions, whatever particle sizes. The lubricant addition induced a positive interaction with Vivapur of greater particle size distribution (200, 102 and 101) favouring higher particle adhesivity, while it maintained unaltered that of Vivapurs of lower particle size (105 and 99). Definitely, when cohesive forces are predominant (Vivapur 105 and 99), the establishment of adhesive bonds between nitrofurantoine and Vivapur remain unnoticed; on the contrary, when cohesion bonds between microcrystalline cellulose particles are weakened by the presence of magnesium stearate, the existence of adhesion bonds between particles of different nature is in evidence, leading to a positive interaction. PMID- 15337443 TI - Synthesis of oligogalacturonates conjugated to BSA. AB - The synthesis of three oligogalacturonates with an aldehyde spacer attached at the reducing end is described. Trigalacturonates alpha-d-GalpA-(1-->4)-alpha-d GalpA-(1-->4)-alpha-d-GalpA-(1-->O(CH(2))(7)CHO and alpha-d-GalpA(Me)-(1-->4) alpha-d-GalpA(Me)-(1-->4)-alpha-d-GalpA(Me)-(1-->O(CH(2))(7)CHO as well as hexagalacturonate alpha-d-GalpA-(1-->4)-[alpha-d-GalpA-(1-->4)](4)-alpha-d-GalpA (1-->O(CH(2))(7)CHO are prepared by stepwise coupling of galactose units followed by oxidation of the 6-positions. The alpha-linkages are formed by employing n pentenyl galactosides as glycosyl donors and N-iodosuccinimide/triethylsilyl triflate as the promoter. Deprotection furnishes the three target oligogalacturonates, which are subsequently linked to bovine serum albumin by reductive amination. These neoglycoproteins will serve as immunogens for generation of new antibodies that can be used for localization and characterization of pectin in plants. PMID- 15337444 TI - Formation of 3-hexuloses in aldol reactions, analysis of the products as their O isopropylidene derivatives by GC-MS. AB - A method for analysis of mixtures of 3-hexuloses by gas chromatography mass spectrometry of their di-O-isopropylidene derivatives has been elaborated. The origin of characteristic fragment ions in the mass spectra is suggested on the basis of the spectra of d(12) analogues, obtained by acetonation with acetone d(6) and on MS/MS investigations. The method has been applied to product mixtures from aldol reactions between glycero-tetrulose and glycolaldehyde and between 2 pentuloses and formaldehyde. An interesting result is the formation of ribo-3 hexulose with a high degree of stereoselectivity in alkali catalysed reaction between erythro-2-pentulose and formaldehyde. PMID- 15337445 TI - Novel perfluoroalkylated derivatives of D-galactopyranose and xylitol for biomedical uses. Hemocompatibility and effect on perfluorocarbon emulsions. AB - 6-O-(4,4,5,5,6,6,7,7,7-Nonafluoro-2-hydroxyheptyl)-, 6-O (4,4,5,5,6,6,7,7,8,8,9,9,9-tridecafluoro-2-hydroxynonyl)-, and 6-O (4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoro-2-hydroxyundecyl)-d galactopyranose (9, 10, and 11, resp.) were prepared by a two-step synthesis including the reaction of 1,2:3,4-di-O-isopropylidene-alpha-d-galactopyranose with 2-[(perfluoroalkyl)methyl]oxiranes under catalysis with BF(3).Et(2)O. Similarly, 1-O-(4,4,5,5,6,6,7,7,7-nonafluoro-2-hydroxyheptyl)-, 1-O (4,4,5,5,6,6,7,7,8,8,9,9,9-tridecafluoro-2-hydroxynonyl)-, 1-O (4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoro-2-hydroxyundecyl)-dl xylitol (18, 19, and 20, resp.) were prepared by a two-step synthesis from the corresponding 1,2:3,4-di-O-isopropylidene-dl-xylitol. Most of the both types of fluoroalkylated carbohydrate derivatives 9-11 and 18-20 generally displayed very low level of hemolytic activity and excellent co-emulsifying properties on testing on perfluorodecalin-Pluronic F-68 microemulsions. PMID- 15337446 TI - Synthesis and structure determination of some nonanomerically C-C-linked serine glycoconjugates structurally related to mannojirimycin. AB - The Bucherer-Bergs reaction of methyl 2,3-O-isopropylidene-alpha-d-lyxo hexofuranosid-5-ulose gave (4'S)-4'-carbamoyl-4'-[methyl (4R)-2,3-O isopropylidene-beta-l-erythrofuranosid-4-C-yl]-oxazolidin-2'-one instead of expected hydantoins. A mixture of hydantoins--(5'R)-triphenylmethoxymethyl-5' [methyl (4R)-2,3-O-isopropylidene-beta-l-erythrofuranosid-4-C-yl]-imidazolidin 2',4'-dione and (5'S)-triphenylmethoxymethyl-5'-[methyl (4R)-2,3-O-isopropylidene beta-l-erythrofuranosid-4-C-yl]-imidazolidin-2',4'-dione was obtained from the 5 ulose having protected primary OH group at C-6. The 4'-S configuration of 2 as well as 5'-S configuration of (5'S)-hydroxymethyl-5'-[methyl (4R)-2,3-O isopropylidene-beta-l-erythrofuranosid-4-C-yl]-imidazolidin-2',4'-dione (9) was confirmed by X-ray crystallography. Corresponding alpha-amino acid--methyl (5S)-5 amino-5-C-carboxy-5-deoxy-alpha-d-lyxo-hexofuranoside (alternative name: 2 [methyl (4R)-beta-l-erythrofuranosid-4-C-yl]-l-serine) (11) was obtained from the hydantoin 9 by acid hydrolysis of the isopropylidene and trityl groups followed by basic hydrolysis of the hydantoin ring. Analogous derivatives with 5-R configuration, formed in a minority, were also isolated and characterised. PMID- 15337447 TI - Catalytic ceric ammonium nitrate mediated synthesis of 2-deoxy-1-thioglycosides. AB - Synthesis of 2-deoxy-1-thioglycosides from glycals, mediated by catalytic amounts of ceric ammonium nitrate is reported. Apart from the 2-deoxy-1-thioglycosides, formation of the 2,3-unsaturated enose, corresponding to the Ferrier product, is also observed, especially for the glucal substrates. A radical oxocarbenium ion and a thiolate intermediates are most likely to mediate the reaction. Upon synthesis of 2-deoxy-1-thioglycosides, few representative glycosylation reactions with both aglycosyl and glycosyl acceptors were performed and alpha-anomeric 2 deoxy glycosides were obtained exclusively. PMID- 15337448 TI - Synthesis of novel ammonium and selenonium ions and their evaluation as inhibitors of UDP-galactopyranose mutase. AB - The syntheses of two ammonium salts of 1,4-dideoxy-1,4-imino-d-galactitol containing erythritol sulfate side chains are described. The parent compound is a known inhibitor of the enzyme UDP-galactopyranose mutase (UDP-galactopyranose furanomutase, E.C. 5.4.99.9), which is responsible for the conversion of UDP galactopyranose into UDP-galactofuranose and is presumably protonated in its active form. The side chain was chosen because it is present in a known sulfonium ion alpha-glucosidase inhibitor, salacinol. The syntheses of the selenonium analogues derived from 1,4-dideoxy-1,4-seleno-d-galactitol are also described. The synthetic strategy in the syntheses of all four salts involved the nucleophilic attack of a protected derivative of the alditol at the least hindered carbon of 2,4-O-benzylidene d- or l-erythritol-1,3-cyclic sulfate to give adducts that were subsequently deprotected. The importance of different protecting groups used in the various syntheses is also highlighted. Enzyme inhibition assays carried out on these compounds, and the corresponding sulfonium ions synthesized previously, show that they are poor inhibitors of UDP galactopyranose mutase. PMID- 15337449 TI - Structural characterization of a 2-O-acetylglucomannan from Dendrobium officinale stem. AB - A heteropolysaccharide obtained from an aqueous extract of dried stem of Dendrobium officinale Kimura and Migo by anion-exchange chromatography and gel permeation chromatography, was investigated by chemical techniques and NMR spectroscopy, and is demonstrated to be a 2-O-acetylglucomannan, composed of mannose, glucose, and arabinose in 40.2:8.4:1 molar ratios. It has a backbone of (1-->4)-linked beta-d-mannopyranosyl residues and beta-d-glucopyranosyl residues, with branches at O-6 consisting of terminal and (1-->3)-linked Manp, (1-->3) linked Glcp, and a small proportion of arabinofuranosyl residues at the terminal position. The acetyl groups are substituted at O-2 of (1-->4)-linked Manp and Glcp. The main repeating unit of the polysaccharides is reported. PMID- 15337450 TI - Multinuclear NMR study of the complexes of 6-phospho-D-gluconic acid with W(VI) and Mo(VI). AB - Multinuclear ((1)H, (13)C, (17)O, (31)P, (95)Mo, (183)W) magnetic resonance spectroscopy (1D and 2D) has been used to show that 6-phospho-d-gluconic acid forms three complexes with tungsten(VI) and six complexes with molybdenum(VI) in aqueous solution, depending on pH and concentration. Two isomeric 1:2 (metal ligand) complexes are detected both with tungstate(VI) and molybdate(VI), having MO(2)(2+) centres and involving the carboxylate and the adjacent OH groups in addition to one 2:1 (metal-ligand) complex possessing a M(2)O(5)(2+) centre, with the ligand being coordinated by the carboxylate group and the three consecutive OH groups in positions 2, 3 and 4. Molybdate(VI) forms three additional species, which are not detected with tungstate. One of them is a 2:1 complex with a Mo(2)O(5)(2+) centre, with the ligand being tetradentate via O-3, O-4, O-5 and the phosphate group. The other two are 12:4 species, which can be seen as two 1:2 complexes bound together in a ring through two diphosphomolybdate moieties each derived from heptamolybdate by inclusion of two phosphate groups from the ligands. PMID- 15337451 TI - Synergistic interactions between the genetically modified bacterial polysaccharide P2 and carob or konjac mannan. AB - Rheological studies have confirmed that the bacterial polysaccharide P2, a genetically modified variant of the Acetobacter xylinum polysaccharide acetan, undergoes synergistic gelation with either of the plant polysaccharides carob or konjac mannan. X-ray fibre diffraction data shows that P2 can form a 5-fold helical structure of pitch 4.7nm and an axial rise per disaccharide repeat of 0.92nm. Optical rotation data demonstrate that P2 undergoes a coil-helix transition in solution and that deacylation enhances the stability of the helical structure in solution. Studies made on mixtures prepared at different temperatures and ionic strengths suggest that denaturation of the P2 helix favours interaction and gelation. Deacetylation of P2 enhances gelation. X-ray diffraction data for oriented fibres prepared from deacetylated P2-konjac mannan mixed films reveal a 6-fold helical structure of pitch 5.54nm with an axial rise per disaccharide repeat also of 0.92nm. This mixed helix provides direct evidence for binding between the two polysaccharides. P2 contains two sites of acetylation: one on the backbone and one on the sidechain. The former site of acetylation inhibits helix formation for P2. It is suggested that this site of acetylation also inhibits formation of the mixed helix, explaining the enhanced gelation of mixtures on deacetylation. PMID- 15337452 TI - The structure of the phosphorylated carbohydrate backbone of the lipopolysaccharide of the phytopathogen bacterium Pseudomonas tolaasii. AB - A novel core-lipid A backbone oligosaccharide was isolated and identified from the lipopolysaccharide fraction of the mushrooms pathogen bacterium Pseudomonas tolaasii. The oligosaccharide was obtained by alkaline treatment of the lipopolysaccharide fraction. Since the repeating unit of the O-antigen contained one residue of -->4)-alpha-l-GulpNAcAN, the hydrolysis was accompanied by beta elimination on this residue and following depolymerization, producing a mixture of oligosaccharides. The complete structural elucidation showed the presence of a single core glycoform and was achieved by chemical analysis and by (1)H, (31)P, and (13)C NMR spectroscopy applying various 1D and 2D experiments. [structure: see text]. All sugars are alpha-d-pyranoses, if not stated otherwise. Hep is l glycero-d-manno-heptose, Kdo is 3-deoxy-d-manno-oct-2-ulosonic acid, P is phosphate. QuiN and DeltaGulNA are present in nonstoichiometric amount. PMID- 15337453 TI - Structural investigation of a water-soluble glucan from an edible mushroom, Astraeus hygrometricus. AB - A water-soluble glucan, Fraction I, was isolated from the aqueous extract of the fruit bodies of the mushroom Astraeus hygrometricus. On the basis of total hydrolysis, methylation analysis, periodate oxidation, and NMR studies ((1)H, (13)C, 2D-COSY, TOCSY, NOESY, and HSQC), the structure of the repeating unit of the glucan is determined as:This glucan shows strong splenocyte activation. PMID- 15337454 TI - Refinement of the structures of cell-wall glucans of Schizosaccharomyces pombe by chemical modification and NMR spectroscopy. AB - Alkali extraction and methylation analyses in the 1970s revealed that the cell walls of the yeast Schizosaccharomyces pombe contain a (1-->3)-alpha-d-glucan, a (1-->3)-beta-d-glucan, a (1-->6)-beta-d-glucan, and a alpha-galactomannan. To refine the structures of these polysaccharides, cell-wall glucans of S. pombe were extracted, fractionated, and analyzed by NMR spectroscopy. S. pombe cells were treated with 3% NaOH, and alkali-soluble and insoluble fractions were prepared. The alkali-insoluble fraction was treated with 0.5M acetic acid or Zymolyase 100T to yield an alkali-insoluble, acetic acid-insoluble fraction, an alkali-insoluble, Zymolyase-insoluble fraction, and an alkali-insoluble, Zymolyase-soluble fraction. (13)C NMR and 2D-NMR spectra disclosed that the cell wall of S. pombe is composed of three types of glucans, specifically, a (1-->3) alpha-d-glucan, a (1-->3)-beta-d-glucan, which may either be linear or slightly branched, and a highly branched (1-->6)-beta-d-glucan, in addition to alpha galactomannan. The highly branched (1-->6)-beta-d-glucan was identified by selective periodate degradation of side-chain glucose as a highly (1-->3)-beta branched (1-->6)-beta-d-glucan with more branches than that of Saccharomyces cerevisiae. Flexibility of these polysaccharides in the cell wall was analyzed by (13)C NMR spectra in D(2)O. The data collectively indicate that (1-->3)-alpha- and (1-->3)-beta-d-glucans are rigid and contribute to the cell shape, while the highly branched (1-->6)-beta-d-glucan and alpha-galactomannan are flexible. PMID- 15337455 TI - Melting behaviour of D-sucrose, D-glucose and D-fructose. AB - The melting behaviour of d-sucrose, d-glucose and d-fructose was studied. The melting peaks were determined with DSC and the start of decomposition was studied with TG at different rates of heating. In addition, melting points were determined with a melting point apparatus. The samples were identified as d sucrose, alpha-d-glucopyranose and beta-d-fructopyranose by powder diffraction measurements. There were differences in melting between the different samples of the same sugar and the rate of heating had a remarkable effect on the melting behaviour. For example, T(o), DeltaH(f) and T(i) (initial temperature of decomposition) at a 1 degrees Cmin(-1) rate of heating were 184.5 degrees C, 126.6Jg(-1) and 171.3 degrees C for d-sucrose, 146.5 degrees C, 185.4Jg(-1) and 152.0 degrees C for d-glucose and 112.7 degrees C, 154.1Jg(-1) and 113.9 degrees C for d-fructose. The same parameters at 10 degrees Cmin(-1) rate of heating were 188.9 degrees C, 134.4Jg(-1) and 189.2 degrees C for d-sucrose, 155.2 degrees C, 194.3Jg(-1) and 170.3 degrees C for d-glucose and 125.7 degrees C, 176.7Jg(-1) and 136.8 degrees C d-fructose. At slow rates of heating, there were substantial differences between the different samples of the same sugar. The melting point determination is a sensitive method for the characterization of crystal quality but it cannot be used alone for the identification of sugar samples in all cases. Therefore, the melting point method should be validated for different sugars. PMID- 15337456 TI - Role of CH/pi interactions in substrate binding by Escherichia coli beta galactosidase. AB - Interactions between carbohydrates and aromatic amino-acid residues are often observed in structures of carbohydrate-protein complexes. They are characterized by an orientation of the pyranose or furanose ring parallel with the aromatic ring of amino-acid residues. An important role in the formation of these complexes is supposed to be played by CH/pi interactions. This paper presents an ab initio quantum chemistry study of CH/pi interactions between beta galactosidase from E. coli and its substrates and products. The energy stabilizing the interaction between Trp999 residue and substrate bound in the shallow binding mode was calculated at the MP2/6-31+G(d) level as 5.2kcalmol(-1) for the glucose moiety of allolactose, 2.4kcalmol(-1) for the galactose moiety of allolactose and 5.0kcalmol(-1) for the glucose moiety of lactose. The energy stabilizing the interaction between Trp568 residue and galactose in the deep binding mode was calculated as 2.7kcalmol(-1). Interaction energies at the HF/6 31+G(d) and B3LYP/6-31+G(d) levels were small or repulsive; therefore, highly correlated ab initio methods were necessary to study these interactions. These unexpectedly strong interactions give a rationale for allolactose formation and illustrate the role of the Trp999 residue. In addition, this illustrates the importance of CH/pi interactions for the function of carbohydrate-binding proteins and carbohydrate-processing enzymes. PMID- 15337457 TI - Pectin methylesterases: sequence-structural features and phylogenetic relationships. AB - Pectin methylesterases (PMEs) are enzymes produced by bacteria, fungi and higher plants. They belong to the carbohydrate esterase family CE-8. This study deals with comparison of 127 amino acid sequences of this family containing the five characteristic sequence segments: 44_GxYxE, 113_QAVAL, 135_QDTL, 157_DFIFG, 223_LGRPW (Daucus carota numbering). Six strictly conserved residues (Gly44, Gly154, Asp157, Gly161, Arg225 and Trp227) and six conservative ones (Ile39, Ser86, Ser137, Ile152, Ile159 and Leu223) were identified. A set of 70 representative PMEs was created. The sequences were aligned and the evolutionary tree based on the alignment was calculated. The tree reflected the taxonomy: the fungal and bacterial PMEs formed their own clusters and the plant enzymes were grouped into eight separate clades. The plant PME from Vitis riparia was placed in a common clade with fungi. Three plant clades (Plant 1, 2 and 3) were relatively homogenous reflecting high degree of mutual sequence identity. The clade Plant 4 contained PMEs from flower parts (mostly form pollen) and was heterogenous, like the clades Plant 1a and 2a, which moreover exhibit an intermediate character. The clades Plant X1 and X2 were situated in the tree close to microbial clades and represented atypical plant PMEs. Taking into account the remaining plant PMEs, an expanded plant alignment and tree (with most Arabidopsis thaliana and Oryza sativa enzymes), were prepared. An exclusive Arabidopsis alignment and tree indicated the existence of a new plant clade X3. In the pre pro region of most plant enzymes a longer conserved segment containing basic dipeptide, R(K)/R(K), that precedes the N-terminal end of PME was revealed. This was not observed in the clade Plant X1 and majority of the clade Plant X2. This study brings further the description of occurrence of potential glycosylation sites in pre pro sequences and in mature enzymes as well as important amino acid residues, such as aspartates, cysteines, histidines and other aromatic residues (Tyr, Phe and Trp), with discussion of their possible function in the activity of PMEs. PMID- 15337458 TI - Evaluation of sulfated fungal beta-glucans from the sclerotium of Pleurotus tuber regium as a potential water-soluble anti-viral agent. AB - Six water-insoluble fractions of fungal beta-glucans extracted by hot alkali (TM8 1 to TM8-6) from the sclerotia of Pleurotus tuber-regium (PTR) having different molecular weights (M(w)) were sulfated to give their corresponding water-soluble derivatives (S-TM8-1 to S-TM8-6) with the degree of sulfation (DS) ranging from 1.14 to 1.74. The in vitro anti-viral activities of the native beta-glucans (TM8s) and their sulfated derivatives (S-TM8s) were evaluated by the cytopathic effect assay (CPE) and the plaque reduction assay (PRA) against four kinds of viruses, including herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), respiratory syncytial virus (RSV), and influenza A virus (Flu A). Although TM8s were inactive in inhibiting the viral replication in cell cultures, the S-TM8 fractions with the defined M(w) range had potent anti-viral activity against HSV-1 and HSV-2 as shown by the CPE assay. The PRA results suggested that S-TM8 fractions seemed to exert their anti-viral effect by binding to the viral particles, preventing the latter from infecting the host cells. It was plausible that the negative charges on the polymer chain of S-TM8 could interact with the positively charged glycoproteins on the surface of HSV, minimizing the interaction between the HSV and the negatively charged host cells. The anti-viral activity of the S-TM8s might also be explained by their more extended chain conformation in solution due to an increase in one of their molecular parameter, persistence length (q), as compared to the native TM8s. The potential use of S TM8s as a water-soluble anti-HSV agent is discussed. PMID- 15337459 TI - Synthesis of fagopyritols A1 and B1 from D-chiro-inositol. AB - Fagopyritol A1 (3-O-alpha-d-galactopyranosyl-d-chiro-inositol) and fagopyritol B1 (2-O-alpha-d-galactopyranosyl-d-chiro-inositol) have been synthesized by glycosylation of the diequatorial diol 1,4,5,6-tetra-O-benzoyl-d-chiro-inositol, readily obtained from d-chiro-inositol, with 2,3,4,6-tetra-O-benzyl-d galactopyranosyl trichloroacetimidate. PMID- 15337460 TI - The revised NMR chemical shift data of carrageenans. AB - A new set of (13)C and (1)H NMR chemical shifts of most common carrageenan types is given relative to DSS as the internal standard according to the IUPAC recommendations. Moreover, the chemical shifts of characteristic signals for pyruvate acetal and floridean starch are reported. Additionally, chemical shifts of common internal standards, such as methanol, DMSO and acetone, were measured at different temperatures and pH values. PMID- 15337461 TI - Isolation of diferulic bridges ester-linked to arabinan in sugar beet cell walls. AB - After degradation of sugar beet cell walls with Driselase and fractionation of the solubilised products by hydrophobic interaction chromatography, a dehydrodiferuloylated oligoarabinan was isolated. Its structure was assigned to two dimers of (1-->5)-linked arabinose units esterified by a central 8-O-4' ferulic dimer. These results provide the first direct evidence that pectic arabinans in sugar beet cell walls may be covalently cross-linked through dehydrodiferulates. PMID- 15337462 TI - Influence of alkali-freezing treatment on the solid state structure of chitin. AB - Chitin was soaked in frozen sodium hydroxide for further modification. The solid state structural changes of the resulting chitin powders were studied by X-ray diffraction (XRD) and infrared spectroscopy measurements. During the alkali freezing process, the crystal space parameters of chitin changed, and the order of the hydrogen bonds of chitin was modified. The study explains how the treatment is beneficial for further modification of chitin. The crystallinity of chitin was assessed according to XRD results. It decreased by half after the alkali-freezing treatment for the first 3 days, and then a few changes happened on the following days while the crystallinity of chitin was kept in the scale of 13.5-18%. That means that the alkali-freezing treatment of chitin for several days is suitable and sufficient. PMID- 15337463 TI - Enzymatic liberation of lycotetraose from the Solanum glycoalkaloid alpha tomatine. AB - The branched tetrasaccharide, O-beta-d-glucopyranosyl-(1-->2)-O-[beta-d xylopyranosyl-(1-->3)]-O-beta-d-glucopyranosyl-(1-->4)-d-galactose (lycotetraose) is a key constituent of many biologically interesting natural products. Described herein is a convenient enzymatic preparation of lycotetraose from the readily available Solanum glycoalkaloid alpha-tomatine. The preparation makes use of the recombinant endo-glycosidase, tomatinase, from the plant pathogen Fusarium oxysporum f. sp. lycopersici. PMID- 15337464 TI - Human serum IgA1 is substituted with up to six O-glycans as shown by matrix assisted laser desorption ionisation time-of-flight mass spectrometry. AB - The micro-heterogeneity of human serum IgA1 results from variable O-glycan substitutions in the 'hinge region' of the molecule and this O-glycosylation may be altered in a number of medical conditions. This micro-heterogeneity has been monitored by analysis of IgA1-derived tryptic O-glycopeptides using matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-ToF MS) analysis. With ammonium citrate-trihydroxyacetophenone matrix, individual compositional glycoforms have been baseline resolved in more than 70 samples and these spectra revealed for the first time that, in addition to expected substitution with 3,4 and 5 GalNAcs, a sixth GalNAc substitution was also present in the hinge region of the molecule. The spectra obtained from subsequent exoglycosidase-treated samples confirmed hexa-O-substitution. Following endoprotease digestions of the exoglycosidase treated samples, possible locations for the sixth GalNAc were indicated from further MALDI-ToF-MS analysis. Hexa substitution accounts for around 5-10% the glycoforms. This is, we believe, the first report of hexa-O-substitution with GalNAc of human serum IgA1. PMID- 15337465 TI - G-protein beta3 subunit gene C825T polymorphism in patients with vesico-ureteric reflux. AB - The C825T polymorphism in the GNB3 gene encoding a beta3 subunit from heterotrimeric G-proteins correlates strongly with the variation in activity of the G-proteins. It has so far been associated with a variety of medical conditions, but has not been tested for association with vesico-ureteric reflux (VUR). Primary VUR is a condition of genetic origin that appears to be inherited in an autosomal dominant mode, but with reduced penetrance. The constitutional change in G-protein-mediated cell signaling associated with the C825T polymorphism might be one of the factors that participate in the development of VUR by modifying the effect of still unknown mutated gene(s). A significant difference in genotype frequencies (chi(2) = 7.38, P = 0.025, df = 2) was observed between patients with primary VUR (33 CC homozygotes, 40 CT heterozygotes, 12 TT homozygotes) and healthy controls with no medical record of reflux (114 CC homozygotes, 88 CT heterozygotes, 18 TT homozygotes). This result suggests that the C825T polymorphism of the GNB3 gene might be associated with the development of VUR. PMID- 15337466 TI - Androgen receptor gene CAG repeats length in fertile and infertile Tunisian men. AB - Several reports implicated a relation between the trinucleotide (CAG) repeat length in the androgen receptor (AR) gene and male infertility. But such result was not reproduced in others. To test this hypothesis, we investigated the number of (CAG) repeats in the AR gene among two groups of infertile (n = 129) and fertile Tunisian men (n = 98), using polymerase chain reaction (PCR) targeting the AR CAG repeat tract, followed by electrophoresis on polyacrylamide gel (6%). For statistical analysis we used Student, Kolmogorov-Smirnov (KS) and chi(2) tests. Significance was reached when P < 0.05. No statistically significant difference in the mean length of the CAG repeat was found between infertile and control groups (P = 0.47). Moreover, using KS test, we have not found a difference in the distribution of allele frequencies between infertile and controls (D(obs) = 0.046 < D(crit) = 0.180). We also did not found a statistically significant relationship between the size of the CAG repeat and impaired sperm production in Tunisian population. Our results may be attributed to the high probability that infertile males may represent a heterogeneous group with respect to the causes of defective spermatogenesis. PMID- 15337467 TI - Auricular mild errors of morphogenesis: epidemiological analysis, local correlations and clinical significance. AB - BACKGROUND: The mild errors or morphogenesis (MEMs) are well known and accepted markers of alterations in embryonic development with predictive value in identification of major malformations, specific genetic syndromes, metabolic and psychiatric disease and childhood malignancy. OBJECTIVE: The goal of this study was to assess the contribution of auricular MEMs as part of total MEMs in an effort to study the factors influencing the different potential informative value of different types of MEMs and their variability with perinatal factors. METHOD: Three thousand one hundred and seven consecutive born neonates were screened for auricular and non-auricular MEMs, inregistered concomitantly with major malformations and postural defects. The study was accomplished by our specially designed computerized program in a relatively large nonhomogeneous ethnic population, in the metropolitan area of Haifa, Israel. RESULTS: The general prevalence of auricular MEMs was 43.1%; the most frequent pathogenetic type was the postural one. Significantly higher rates of auricular MEMs were associated with male sex, small- and large-for-gestational age, IVF pregnancy, triplet pregnancy, maternal diabetes and parental consanguinity. CONCLUSION: We conclude that the presence, number, and association or concomitance of auricular MEMs in the same neonate may have important clinical, diagnostic, pathogenetic, screening, and therapeutic value. PMID- 15337468 TI - Allelic variations at the haploid TBX1 locus do not influence the cardiac phenotype in cases of 22q11 microdeletion. AB - Microdeletion at the 22q11 locus is characterised by a high clinical variability. Congenital heart defects (CHD) are the most life-threatening manifestations of the syndrome and affect approximately 50% of patients carrying the deleted chromosome 22. The causes of this phenotype variability remain unknown although several hypotheses have been raised. It has been suggested that allelic variations at the haploid locus could modify the phenotypic expression. Regarding this hypothesis, TBX1 was thought to be a major candidate to the cardiac phenotype or its severity in patients carrying the 22q11 microdeletion. A mutational screening was performed in this gene, in a series of 39 deleted patients, with and without CHD. The results indicate that mutations in TBX1 are not likely to be involved in the cardiac phenotype observed in del22q11 patients. PMID- 15337469 TI - Clinical findings and cytogenetic analysis of small supernumerary ring chromosomes 7: report of two new cases. AB - Two new patients, mosaic for a small supernumerary ring chromosome 7 are described. There are only seven published reported concerning supernumerary ring chromosome 7 and we reviewed the previously reported cases in an attempt to establish genotype-phenotype correlations, which are particularly important for genetic counselling and clinical genetics. Our first case was a 20 months old girl who was referred for a mild motor developmental delay, an asymmetric facial appearance, a plagiocephaly and a short nose with anteverted nostrils. Our second case was a 9 years old boy who was referred for a IQ at the lower end of the normal range (? 80), obesity, hyperactivity and some dysmorphic features including hypertelorism and down slanting palpebral fissures. In both cases, chromosome analysis after G and R banding and FISH showed a small ring chromosome 7 in respectively 76% and 50% of consecutively scored metaphases. Both ring chromosomes were labelled by FISH using the Williams Syndrome locus probe (Elastin Gene D7S486). Comparison between these two cases and previously published cases allowed to delineate frequent clinical findings. A mild mental retardation was found in the majority of patients. which is an important data for genetic counselling. PMID- 15337470 TI - Maternal uniparental disomy 14 dissection of the phenotype with respect to rare autosomal recessively inherited traits, trisomy mosaicism, and genomic imprinting. AB - The phenotype of maternal uniparental disomy of chromosome 14 (upd(14)mat) is characterized by pre and postnatal growth retardation, early onset of puberty, joint laxity, motor delay, and minor dysmorphic features of the face, hands, and feet. Based on a clinical analysis of 24 cases extracted from the literature the phenotype of upd(14)mat was dissected with respect to each symptom's most likely primary causative: trisomy mosaicism, rare autosomal recessively inherited traits, and the impact of known imprinted genes located on chromosome 14q32. As a result, primary factors are confined placental mosaicism for prenatal growth retardation and one or more imprinted genes, which contribute to the reduced final height by accelerated skeletal maturation. As a secondary effect the latter might also cause early onset of puberty. Other secondary effects might be postnatal adaptation problems associated with neurological deficits such as muscular hypotonia due to premature delivery and reduced birthweight and most dysmorphic features as a consequence of subtle skeletal abnormalities and muscular hypotonia. Considering the rarity of traits such as cleft palate, trisomy mosaicism in the fetus is more likely causative than homozygosity of autosomal recessively inherited mutations. Totally, the variable phenotype of upd(14)mat is mainly the consequence of trisomy mosaicism and genomic imprinting. Rare traits might be due to homozygosity of autosomal recessively inherited mutations. PMID- 15337471 TI - A case of macrocephaly-cutis marmorata telangiectatica congenita and review of neuroradiologic features. AB - Macrocephaly-cutis marmorata telangiectatica congenita (M-CMTC) is characterized by macrocephaly, cutis marmorata, capillary malformations, toe syndactily, joint laxity and pre-natal overgrowth. Cerebral abnormalities might also be seen. We reported cerebral magnetic resonance imaging (MRI) findings of a case with M CMTC, who had giant atrial septal aneurysm and atrial septal defect. Cerebral alterations determined by MRI were bilateral prominent lateral ventricles, bilateral cortical dysplasia, cavum septi pellucidum cyst and calvarial hemangioma. At 17th day of his life he suddenly developed cardiorespiratory arrest and died. PMID- 15337472 TI - Prader-Willi syndrome with an unusually large 15q deletion due to an unbalanced translocation t(4;15). AB - Prader-Willi syndrome (PWS) is a neurobehavioral disorder caused by deletions in the 15q11-q13 region, by maternal uniparental disomy of chromosome 15 or by imprinting defects. Structural rearrangements of chromosome 15 have been described in about 5% of the patients with typical or atypical PWS phenotype. An 8-year-old boy with a clinical diagnosis of PWS, severe neurodevelopmental delay, absence of speech and mental retardation was studied by cytogenetic and molecular techniques, and an unbalanced de novo karyotype 45,XY,der(4)t(4;15)(q35;q14),-15 was detected after GTG-banding. The patient was diagnosed by SNURF-SNRPN exon 1 methylation assay, and the extent of the deletions on chromosomes 4 and 15 was investigated by microsatellite analysis of markers located in 4qter and 15q13-q14 regions. The deletion of chromosome 4q was distal to D4S1652, and that of chromosome 15 was located between D15S1043 and D15S1010. Our patient's severely affected phenotype could be due to the extent of the deletion, larger than usually seen in PWS patients, although the unbalance of the derivative chromosome 4 cannot be ruled out as another possible cause. The breakpoint was located in the subtelomeric region, very close to the telomere, a region that has been described as having the lowest gene concentrations in the human genome. PMID- 15337473 TI - A familial complex chromosome translocation resulting in duplication of 6p25. AB - We report on a girl with psychomotor retardation, severe speech developmental delay and mild dysmorphic features. Molecular cytogenetic analysis showed that the patient was carrier of an insertion (6)(p22.5-->22.4) in chromosome 12. Analysis of the chromosomes of the mother revealed the presence of a complex chromosomal rearrangement. In addition to the insertion (6)(p22.5-->22.4) in chromosome 12 and a pericentric inversion in chromosome 12, the 6p subtelomeric region was absent in the mother. This is, to our knowledge, the smallest pure duplication of chromosome 6p as well as the smallest cryptic subtelomeric 6pter deletion thus far reported. PMID- 15337474 TI - Chromosome 10p deletion in a patient with hypoparathyroidism, severe mental retardation, autism and basal ganglia calcifications. AB - Chromosome 10p terminal deletions have been associated with a DiGeorge like phenotype. Haploinsufficiency of the region 10p14-pter, results in hypoparathyroidism, sensorineural deafness, renal anomaly, that is the triad that features the HDR syndrome. Van Esch (2000) identified in a HDR patient, within a 200 kb critical region, the GATA3 gene, a transcription factor involved in the embryonic development of the parathyroids, auditory system and kidneys. We describe a new male patient, 33-year-old, with 10p partial deletion affected by hypocalcemia, basal ganglia calcifications and a severe autistic syndrome associated with mental retardation. Neurologically he presented severe impairment of language, hypotonia, clumsiness and a postural dystonic attitude. A peripheral involvement of auditory pathways was documented by auditory evoked potentials alterations. CT scan documented basal ganglia calcifications. Hyperintensity of the lentiform nuclei was evident at the MRI examination. Renal ultrasound scan was normal. Haploinsufficiency for GATA3 gene was documented with FISH analysis using cosmid clone 1.2. Phenotypic spectrum observed in del (10p) is more severe than the classical DGS spectrum. GATA3 has been found to regulate the development of serotoninergic neurons. A serotoninergic dysfunction may be linked with autism in this patient. PMID- 15337475 TI - Inherited ring chromosome 8 without loss of subtelomeric sequences. AB - We report the first case of inherited ring chromosome 8 syndrome without loss of subtelomeric sequences. The proband is a 6 1/2-year-old boy with short stature, microcephaly, mild mental retardation, and behavioral problems including hyperactivity and attention deficit. His mother presented the same physical features but intelligence was normal. Family history also revealed an uncle and a grandmother, with short stature and microcephaly. Moderate mental retardation was reported in the uncle. Karyotypes and fluorescence in situ hybridization (FISH) analyses were performed on peripheral blood lymphocytes for both child and mother. The child's karyotype was reported as 46,XY,r(8)(p23q24.3)[24]/45,XY, 8[2] and the mother's karyotype 46,XX,r(8)(p23q24.3)[22]/45,XX, 8[2]/47,XX,r(8)(p23q24.3), +r(8)(p23q24.3)[1]. FISH studies showed no deletion of subtelomeric sequences for both child and mother indicating that no or little chromosomal euchromatic material has been deleted. These findings indicate that ring chromosome 8 without loss of subtelomeric sequences can be inherited and that carriers in a same family present with cognitive function ranging from mild mental retardation to normal intelligence. PMID- 15337476 TI - Recurrent proximal 18p monosomy and 18q trisomy in a family with a maternal pericentric inversion of chromosome 18. AB - We report a recurrent partial monosomy of 18p10-->11.2 and proximal partial trisomy of 18q10-->21.3 caused by a maternal pericentric inversion of chromosome 18, involving breakpoints p11.2 and q21q21.3 Based on cytogenetics and FISH analysis, we speculate that the recurrent chromosome abnormality in the proband and in the fetus was the result of a translocation, possibly in a germ cell or germ cell precursor, between the maternal normal 18 and her inverted 18, resulting in maternal germinal mosaicism, i.e. 46,XX,inv(18)/46,XX,t[18;inv(18)][q10;q10]. The unbalanced karyotype of the proband and the fetus is 46,XY,+18,der[18;inv(18)][q10;q10]. To the best of our knowledge, there are no reports of this combination of proximal 18p monosomy and proximal 18q trisomy. The other interesting observation was association of Hirschsprung's disease in the proband. PMID- 15337477 TI - Turner syndrome female with a small ring X chromosome lacking the XIST, an unexpectedly mild phenotype and an atypical association with alopecia universalis. AB - Rearranged X chromosome in Turner syndrome (TS) are generally well tolerated but in cases of ring X chromosomes and of X/autosome translocations the incidence of mental retardation and other congenital abnormalities can be significantly higher. These abnormal phenotypes can be ascribed to failed or partial X inactivation. Here, we report a 10-year-old female who was referred for a cytogenetic analysis because she developed an alopecia universalis. The patient, of normal intelligence, had been found to have traits of TS, especially short stature. A first cytogenetic analysis showed a no mosaic 45,X karyotype. Since, the risk of developing gonadoblastoma in TS patients with mosaicism for a Y derivative chromosome and because association of alopecia universalis and TS is uncommon, fluorescence in situ hybridization (FISH) was performed to search for a second cell population. Our patient was found to have a mosaic 45,X/46,X,+r. FISH analysis using sex chromosome probes permitted us to identify the very small marker as a ring X chromosome, detected in 90% of cells. The ring appeared to be formed almost totally of alphoid sequences with breakpoints in the juxtacentromeric region. The r(X) does not include the XIST locus and may, therefore, not be subject to X-inactivation. Unexpectedly mild phenotype in our patient and its association with alopecia universalis will be discussed. PMID- 15337478 TI - Balanced complex chromosomal rearrangements (BCCR) with at least three chromosomes and three or more breakpoints: report of three new cases. AB - Balanced complex chromosomal rearrangements (BCCR) encompass a heterogeneous group of rare chromosomal aberrations. In this paper, we report three cases of BCCRs. In two the probands were referred for either genetic counseling or prenatal management. One case was ascertained after chromosome analysis performed because of psychiatric manifestations; this was an isolated finding. We also outline the molecular cytogenetic techniques, which were essential in confirming and precisely delineating the BCCRs identified in these patients. In addition the various aspects of genetic counseling for this type of chromosomal rearrangement, highlighting the details particular to each individual case are discussed. We discuss the classification for this type of chromosomal mutation. PMID- 15337479 TI - A sporting chance. PMID- 15337480 TI - Optimum cancer care--an unaffordable goal? PMID- 15337481 TI - Optimum cancer care--an unaffordable goal? PMID- 15337482 TI - Positron-emission tomography in prognostic and therapeutic assessment of lung cancer: systematic review. AB - Positron-emission tomography (PET) with 18-fluorodeoxyglucose has a role in the diagnosis and staging of lung cancer, but is also appealing for assessment of prognosis and treatment. A systematic search of the published work shows good evidence that [(18)F]FDG uptake on PET has independent prognostic value in newly diagnosed non-small-cell lung cancer. PET is a sensitive method of measuring the biological effects of anticancer therapy, but until better standardisation and large-scale experience is available, it should only be used for additional assessments of early response in clinical trials. Further studies are needed to define the role of [(18)F]FDG-PET in restaging after induction therapy in multimodality approaches for locally advanced lung cancer. The assessment of prognosis by [(18)F]FDG-PET is less substantiated in treated lung cancer than in newly diagnosed patients. Good prospective evidence documents the effectiveness of [(18)F]FDG-PET over CT in the correct identification of recurrent lung cancer. PMID- 15337483 TI - Molecular therapy in pancreatic adenocarcinoma. AB - Pancreatic cancer is one of the commonest causes of death from cancer. Despite therapy with surgery, conventional chemotherapy, and radiation, 5-year survival for patients with this diagnosis remains poor. However, advances in the molecular understanding of this malignant disease over the past 5 years might lead to new treatment strategies. Strategies of gene therapy, antiangiogenic treatments, immunotherapy, and signal-transduction inhibition are in preclinical development. This review presents an overview of molecular therapy in pancreatic cancer. PMID- 15337484 TI - Digital gangrene and Raynaud's phenomenon as complications of lung adenocarcinoma. PMID- 15337485 TI - Radiofrequency ablation of liver tumours: systematic review. AB - Thermal ablation by use of radiofrequency energy can be used to achieve necrosis of liver tumours, and increased availability of this technique is leading to more widespread use. Much of the impetus for the use of radiofrequency ablation has come from cohort series that have provided an evidence base for this technique. Here, we give an overview of the current status of radiofrequency ablation for liver tumours, including its physical properties, to assess the characteristics that make this technique applicable in clinical practice. We review the technical development of probe design and summarise current indications and outcomes of reported clinical use. We also provide a profile of side-effects and information on the integration of this technique into the general management of patients with liver tumours. Current evidence suggests that radiofrequency ablation can be done with few side effects; however, although this technique seems to ablate tumours effectively, it should form part of multidisciplinary care for liver cancer. Crucially, the role of radiofrequency ablation in lengthening the survival of patients with liver tumours remains to be assessed. PMID- 15337486 TI - Allogeneic stem-cell transplantation for renal-cell cancer. AB - Metastatic renal-cell carcinoma (RCC) is resistant to chemotherapy, and patients with this disease have a poor outlook. Immunotherapy by use of cytokines and vaccines against tumour antigens has shown encouraging results in a small group of patients. Advances in the understanding of the graft-versus-tumour effect in haematological malignant disorders have led to the use of stem-cell transplantation for treatment of solid-organ malignant diseases such as RCC. Techniques of bone-marrow ablation have been superseded by safer conditioning regimens, with occasional complete remission and partial remission in some patients. Graft-versus-host disease, engraftment failure, and disease progression remain important obstacles to the widespread use of new techniques for metastatic RCC. Here, we summarise important issues surrounding immunotherapy for RCC, the problems encountered with use of immunotherapy, and the present use of non myeloablative techniques for treatment of this disease. PMID- 15337487 TI - The economics of cancer care in the UK. AB - Cancer care accounts for an increasing proportion of global spending on healthcare, driven by an increased incidence caused by ageing populations, greater frequency from better treatments, and changes in care that have made cancer a chronic, controllable illness. The cost of cancer care has three components: direct and easily determined clinical costs (ie, medical costs); extra financial requirement of living with disease for the patient and their family (ie, morbidity costs); and loss of income from the premature death (ie, mortality costs). Effective planning of cancer services needs detailed consideration of the economics of care delivery-an area of research that has so far been lacking outside the USA. Here, we review the literature and attempt to answer key questions on the economics of cancer care, including probable changes in disease burden over the next 20 years, changes in the way costs will be distributed between carers and delivery services, changing patterns of service delivery, and the direct contribution patients will make to treatment costs in terms of co-payments and escalating costs of comorbidity in elderly populations. PMID- 15337488 TI - Clinical and pathological absence of cardiotoxicity after liposomal doxorubicin. PMID- 15337489 TI - . . . and covering up. PMID- 15337490 TI - Arsenic in rice--understanding a new disaster for South-East Asia. PMID- 15337491 TI - Potential of metabolomics as a functional genomics tool. PMID- 15337492 TI - Role of crop physiology in predicting gene-to-phenotype relationships. PMID- 15337493 TI - Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites. PMID- 15337494 TI - One potato, two potato: haplotype association mapping in autotetraploids. PMID- 15337495 TI - Plant responses to hypoxia--is survival a balancing act? PMID- 15337496 TI - All-native DNA transformation: a new approach to plant genetic engineering. PMID- 15337497 TI - The effect of short- versus long-term administration of alpha tocopherol on the survival of random flaps in experimental diabetes mellitus. AB - The effects of short- versus long-term alpha tocopherol administration on oxidative stress and survival of dorsal random flaps were studied in diabetic rats. Seven groups, with 20 rats in each, were constructed: (1) control, (2) noncontrolled diabetes, (3) noncontrolled diabetes+short-term alpha tocopherol, (4) noncontrolled diabetes+long-term alpha tocopherol, (5) insulin treatment, (6) insulin+short-term alpha tocopherol, and (7) insulin+long-term alpha tocopherol. After 3 months of diabetes, dorsal McFarlane flaps were raised. Flap viability and free-radical measurements with histopathological examination were investigated. Mean flap survival in Groups I to VII were 84.0+/-2.2%, 55.0+/ 2.4%, 57.0+/-2.5%, 57.8+/-3.7%, 64.1+/-4.1%, 70.0+/-4.9%, and 77.0+/-6.6%, respectively. Free-radical concentration, as assessed with luminol- and lucigenin enhanced chemiluminiscence, was inversely correlated with flap survival. The results for viability and free-radical concentrations were significant between Groups 1, 2, 5, 6, and 7. Random flaps in diabetic animals showed significantly greater necrosis compared with controls. Among the diabetic animals, group receiving combination of insulin and long-term alpha tocopherol treatment had the greatest flap viability and least tissue free-radical concentration. Histopathological studies showed a hyalinization of arterioles in diabetics with long-term alpha tocopherol treatment protecting the vessel wall. In conclusion, random flaps in experimental diabetes mellitus show greater tissue oxidative stress and necrosis, which is only partially corrected with insulin treatment. Long-term antioxidant supplementation as an adjunct to insulin further lowers the oxidative stress, protects vessel structure and function, and therefore increases flap survival. PMID- 15337498 TI - Lipids and lipoprotein(a) concentrations in Tunisian type 2 diabetic patients; Relationship to glycemic control and coronary heart disease. AB - The aim of this study was to evaluate plasma lipoprotein(a) [Lp(a)] concentrations in Tunisian patients with type 2 diabetes mellitus (DM), to correlate the values with other lipid parameters, and to examine the relationship to glycemic control and coronary heart disease (CHD). Diabetic patients with and without CHD (n=200) had significantly higher levels of Lp(a) (327.94+/-239.93 mg/l) and a greater proportion of elevated (>300 mg/l) Lp(a) concentrations (46%) compared with 100 healthy nondiabetic controls (269.83+/-225.6 mg/l, P<.01, and 26%, P<.01), while there were no statistically significant difference between diabetics without CHD (n=100) and controls. No significant association of Lp(a) with glycemic control (HbAlc or fasting blood glucose) was noted in diabetic patients. Positive correlations were observed between Lp(a) levels and total cholesterol and LDL-C in all diabetic patients and particularly in diabetic men. Male patients with CHD showed significantly higher plasma Lp(a) levels than those without CHD (P=.023), and 57.3% of patients with CHD showed increase (>300 mg/l) Lp(a) compared with 33.3% of patients without CHD. Elevated levels of Lp (a) and abnormal lipid profile in diabetic men suggest their involvement in atherogenesis and subsequent development of CHD. PMID- 15337499 TI - Prevalence of diabetic complications in fibrocalculous pancreatic diabetic patients and type 2 diabetic patients: a cross-sectional comparative study. AB - OBJECTIVE: To determine the prevalence of diabetes-related complications in subjects with fibrocalculous pancreatic diabetes (FCPD) and compare them with subjects with type 2 diabetes mellitus matched for age, sex, and duration of diabetes. METHODS: The study group comprised of 277 FCPD patients and 277 age, sex, and duration of diabetes-matched type 2 diabetic patients. All the study subjects underwent a detailed clinical examination, and fasting blood samples were obtained for biochemical studies. Peripheral Doppler was used for diagnosis of peripheral vascular disease (PVD). Vibratory perception threshold (VPT) was determined using biothesiometry for diagnosis of neuropathy. Diagnosis of coronary artery disease (CAD) was based on medical history and 12-lead resting ECG. Retinal photographs were used for diagnosis of retinopathy using a modified version of Early Treatment Diabetic Retinopathy Study (ETDRS) grading system. RESULTS: FCPD patients had lower body mass index (BMI) (P<.001), systolic blood pressure (P<.0001), diastolic blood pressure (P<.001), serum cholesterol (P<.001), serum triglyceride (P<.001), and serum creatinine (P<.01) but higher glycosylated hemoglobin (P<.001) levels compared to patients with type 2 diabetes. Prevalence of CAD was significantly higher among type 2 diabetic patients (11.9%) compared to FCPD patients (5.1%), P<.003. There was no significant difference in the prevalence of other diabetic complications between the two study groups (type 2 diabetes vs. FCPD: retinopathy-37.2% vs. 30.1%, PVD 4.3% vs. 4.7%, Neuropathy-25.3% vs. 20.9%, Nephropathy-15.0% vs. 10.1%). Multiple logistic regression analysis revealed the following risk factors for diabetes complications among type 2 diabetic subjects-retinopathy: BMI (P=.028), duration of diabetes (P<.001), and glycosylated hemoglobin (P=.026); nephropathy: diastolic blood pressure (P=.016) and glycosylated hemoglobin (P=.040); neuropathy: age (P<.001), duration of diabetes (P=.003), and glycosylated hemoglobin (P=.001). Among subjects with FCPD, systolic blood pressure (P=.013), glycosylated hemoglobin (P=.021), and duration of diabetes (P<.001) were associated with retinopathy; BMI (P=.057), glycosylated hemoglobin (P=.010), and duration of diabetes (P=.024) with nephropathy and age (P=.011) and BMI (P=.010) with neuropathy. CONCLUSION: The prevalence of retinopathy, nephropathy, neuropathy, and PVD was similar among FCPD patients and type 2 diabetic patients, but the prevalence of CAD was lower among FCPD patients. PMID- 15337500 TI - Carnitine deficiency in children and adolescents with type 1 diabetes. AB - Carnitine is essential for the lipid and carbohydrate metabolism, and proper metabolic control in type 1 diabetes has potential impact on long-term complications. The plasma total, free, and acylcarnitine levels in 47 children and adolescents with type 1 diabetes were determined by a radioisotopic assay and compared to the values of a series of anthropometric measurements and metabolic parameters, including blood glycosylated hemoglobin Alc, serum cholesterol and triglycerides, and urine microalbumin levels. Plasma values for total, free, and acylcarnitine were 30.1+/-7.26, 20.0+/-4.50, and 10.2+/-6.47 micromol/l, respectively. Acyl/free carnitine ratio was 0.544+/-0.369. Individuals with type 1 diabetes had significantly lower total and free carnitine levels and significantly higher acyl/free carnitine ratios than controls (P<.001). Plasma total and free carnitine levels were inversely correlated to the duration of diabetes (P=.036 and P=.071, respectively). No statistical relationship was documented between carnitine levels and the remaining anthropometric and metabolic variables. In conclusion, total and free carnitine levels are decreased in children and adolescents with type 1 diabetes. This reduction is time related and may have potential interactions with the long-term complications of type 1 diabetes. Larger studies are required for final conclusions to be drawn on the precise role of carnitine and the possible benefit, if any, of carnitine supplementation in diabetic patients. PMID- 15337501 TI - Sex difference and possible relationship to microvascular complications of serum prorenin levels in type 2 diabetic patients, measured by a novel antibody activating direct enzyme kinetic assay. AB - Prorenin, a precursor of renin, was measured in sera from normal subjects and type 2 diabetic patients, using a newly developed antibody-activating direct enzyme kinetic assay. Levels of prorenin were examined in relationship to diabetic microangiopathies. The levels of antibody-activating direct prorenin (AAD-PR) were approximately 1.5-fold higher than levels of prorenin measured by a conventional enzyme-activating method. AAD-PR levels were significantly higher in males than in females and in diabetic patients than in normal subjects. Moreover, AAD-PR levels were higher in diabetic patients with microalbuminuria and even higher in those with macroalbuminuria. In normoalbuminuric diabetic patients, AAD PR levels were higher in those with retinopathy. Furthermore, a significant positive correlation was seen between the AAD-PR levels and HbA(lc) in normoalbuminuric diabetic subjects without retinopathy. Thus, the determination of circulating serum prorenin measured as AAD-PR is related to glycemia and in type 2 diabetic patients may be a risk marker of diabetic microangiopathy. More studies are necessary to determine whether AAD-PR may actually predict the development of microangiopathy. PMID- 15337502 TI - Reversal of hyperglycemia and diabetic nephropathy: effect of reinstitution of good metabolic control on oxidative stress in the kidney of diabetic rats. AB - Studies have shown that good metabolic control (GC) is beneficial in slowing the progression of nephropathy in diabetes, and if the duration of poor metabolic control (PC) is prolonged before reinstitution of GC, nephropathy is not easily reversed. This study is to identify the biochemical abnormalities that could contribute to the resistance of nephropathy to reverse after establishment of GC in rats. The effect of reinstitution of GC and its duration is evaluated on oxidative stress and nitric oxide (NO) levels in the renal cortex and urine of diabetic rats. The rats were maintained in GC (5% glycated hemoglobin, GHb) soon after or 6 months after induction of hyperglycemia, and were sacrificed 13 months after induction of diabetes. For rats in which GC was initiated soon after induction of diabetes, oxidative stress [as measured by the levels of lipid peroxides (LPOs), 8-hydroxy-2'-deoxyguanosine (8-OHdG), and reduced glutathione (GSH)] and NO in urine and renal cortex were not different from that observed in normal control rats, but when reinstitution of GC was delayed for 6 months after induction of diabetes, oxidative stress and NO remain elevated in both urine and renal cortex. This suggests that hyperglycemia-induced oxidative stress and NO can be prevented if GC is initiated very early, but are not easily reversed if PC is maintained for longer durations. Understanding the mechanisms responsible for this phenomenon could reveal novel means to reverse nephropathy in diabetic patients. PMID- 15337503 TI - Aspirin rectifies calcium homeostasis, decreases reactive oxygen species, and increases NO production in high glucose-exposed human endothelial cells. AB - Aspirin's pharmacological action is mainly related to its property to inhibit prostaglandin synthesis; apart from this, aspirin has some beneficial side effects that are not completely understood, yet. Since aspirin possesses antioxidant properties and antioxidants prevent high d-glucose enhanced endothelial [Ca(2+)](i), we questioned whether aspirin also has an effect on this process as well as on high-glucose-impaired nitric oxide (NO) production. For these purposes, human endothelial cells (HECs) were cultured in normal concentration (5 mM) glucose (NG) or high concentration (33 mM) glucose (HG) and after confluence, exposed for 48 h to HG in the absence or presence of 1 mM aspirin. Then, the [Ca(2+)](i) was measured fluorimetrically using fura-2, NO production was determined by Griess reaction, superoxide anions (O(2)) was evaluated by ferricytochrome c reduction, the intracellular reactive oxygen species (ROS) were evaluated by fluorimetry, and the levels of protein kinase C (PKC) by Western blot. The results showed that HECs exposed to HG displayed: (i) increased [Ca(2+)](i); (ii) enhanced O(2) release; (iii) augmented level of intracellular ROS; and (iv) PKC translocation to the membrane fraction. By comparison, exposure to cells grown in HG to 1 mM aspirin resulted in: (i) a reduction of histamine stimulated [Ca(2+)](i) release to control level and of [Ca(2+)](i) entry by 30%; (ii) a twofold increase in NO production; (iii) a decrease of O(2)(-) accumulation in both culture medium and cell homogenate (by 60.4% and 70%, respectively); (iv) a decline of ROS to the control levels; and (v) a reduction of PKC translocation to the control levels. These data indicate that aspirin corrects the high-glucose-induced changes in cellular Ca(2+) homeostasis and NO production, via a mechanism involving the reduction of the O(2)(-) levels possible by acting on PKC-induced NADPH activity. PMID- 15337504 TI - TGFbeta1 is involved in high glucose-induced accumulation of pericellular chondroitin sulphate in human endothelial cells. AB - High glucose-induced endothelial cell dysfunction is considered to be the main cause of the development of vascular diabetes complications. Cultured endothelial cells exposed to high glucose in vitro demonstrate a variety of alterations, including extracellular matrix (ECM) deposition, growth inhibition, and changes in cell motility. Some of these effects were shown to be mediated by the up regulation of endothelial transforming growth factor-beta1 (TGFbeta1) secretion and activation. We investigated the influence of high glucose on human immortalized endothelial cell line ECV304. According to our data, confluent cells exposed to 30 mM glucose for 48 h secrete the increased amount of total and active TGFbeta1 ( approximately 1.4-fold), and accumulate more chondroitin sulphate (CS) in their conditioned medium, pericellular matrix, and cell layer ( approximately 1.6- to 2.0-fold). By blocking the coupling of CS chains to the core protein with p-nitrophenyl-beta-D-xyloside and by chondroitinase ABC treatment, we demonstrated that the increased accumulation of pericellular CS is accompanied by increased cell attachment to immobilized hyaluronic acid (HA), while the expression of cell surface CD44 remains unaltered. Since the exogenous TGFbeta1 affects ECV304 cells in a similar manner, and anti-TGFbeta1-neutralizing antibody cancels the effect of high glucose, we suggest the involvement of TGFbeta1 in the development of endothelial cell response to high glucose in terms of CS accumulation and cell binding to HA. PMID- 15337505 TI - Tranilast reduces mesenteric vascular collagen deposition and chymase-positive mast cells in experimental diabetes. AB - The mast cell has a central role in the pathogenesis of fibrosis a common feature of diabetic microvascular complications. Increased mast cell numbers have been demonstrated in diabetic nephropathy in association with renal fibrosis, and diabetes acutely increases mast cell infiltration in the mesentery. Antimast cell agents such as tranilast may ameliorate the acute vascular changes in diabetes due to stabilisation of mast cells and/or reduction in mast cell numbers. After 3 weeks of streptozotocin diabetes, light microscopy techniques were used to estimate mesenteric vessel fibrosis and mast cell infiltration. Mast cells were identified by toluidine blue staining and tryptase, chymase and TGF-beta immunohistochemistry in three study groups of rats: control, diabetic and plus tranilast. Diabetes was associated with an increase in both mesenteric vessel fibrosis and mast cell numbers. Administration of tranilast to diabetic rats reduced mesenteric vessel fibrosis and this was associated with a reduction in chymase-positive mast cells. These changes were independent of mast cell TGF-beta and were not associated with a reduction in tryptase-positive mast cells. The amelioration of diabetes-induced vessel fibrosis may be due to a reduction in the liberation of angiotensin II by inhibiting mast cell chymase. PMID- 15337506 TI - Half protons or doubly charged protons? The history of metastable ions. AB - The history of metastable ions is intertwined with that of ions formed by collision-induced dissociation (CID), and frequently the genesis of the two ion types cannot be strictly separated. Originally, metastable ions were considered a curiosity or even a nuisance, being responsible for "humps" in the base line of the recorded mass spectra. In their heyday metastable ions were recognized as having importance for establishing fragmentation sequences and for distinguishing between isomeric ion structures. Today, in many respects the utility of metastable ions has been superseded by a systematic application of CID techniques; yet the evaluation of their shape is still of importance for questions of reaction energies and ion thermochemistry. PMID- 15337507 TI - Long-lived excited electronic states of the dichloroethylene cation isomers probed by charge exchange ionization. AB - Charge exchange technique has been used to detect the presence of long-lived excited electronic states of trans-, cis-, and 1,1-C(2)H(2)Cl(2)(+.). The B states of the three cations which are formed by removal of an electron from an in plane chlorine nonbonding orbital of the corresponding neutrals have been found to have long lifetimes (tens of microseconds or longer). Whether the A states formed by removal of an electron from the other in-plane chlorine nonbonding orbitals are long-lived also can not be determined by the present experiments. Cations in the excited electronic states above the B? states were not detected because of their prompt dissociation following intramolecular relaxation or radiative decay. PMID- 15337508 TI - Delineating mechanisms of dissociation for isomeric heparin disaccharides using isotope labeling and ion trap tandem mass spectrometry. AB - Heparin and heparan sulfate (HS) glycosaminoglycans have been identified as important players in many physiological as well as pathophysiological settings. A better understanding of the biosynthesis and structure of these molecules is critical for further elucidation of their biological function. We have demonstrated the successful use of negative electrospray ionization tandem mass spectrometry in the differentiation of all twelve standard heparin-building blocks, including the potentially important N-unsubstituted disaccharides. Collision induced dissociation of each of the isomeric disaccharides provided unique product ion spectra, useful for identification and quantification of the relative amounts of each isomer present. In the research presented herein, isotopic labeling studies using (18)O and (2)H were used to determine the origins of each of the neutral losses observed in the product ion spectra, and mechanisms of dissociation consistent with the observed data were postulated. The general mechanisms postulated were for the generation of B, Y, and Z ions formed from glycosidic cleavages, as well as A and X ions formed from cross-ring cleavages. The eight isomeric heparin disaccharides all underwent cross-ring cleavage to form (0,2)X(1) and (0,2)A(2) ions, and further experiments suggest that the mechanisms of formation of these ions are through a charge-remote process. The tandem mass spectrometry data presented herein also provide a foundation for further developments towards a practical analysis tool for the structural elucidation of larger, biologically important heparin/HS oligosaccharides by using mass spectrometry. PMID- 15337509 TI - Determination of the glycosylation site of flavonoid monoglucosides by metal complexation and tandem mass spectrometry. AB - Metal complexation and tandem mass spectrometry were used to differentiate C- and O-bonded flavonoid monoglucoside isomers. Electrospray ionization of solutions containing a flavonoid glycoside and a metal salt led to the generation of the key [M(II) (L) (L-H)](+) complexes, where M is the metal ion and L is the flavonoid glycoside. Thirteen flavonoid monoglucosides were examined in combination with Ca(II), Mg(II), Co(II), Ni(II), and Cu(II). Collisional activated dissociation (CAD) of the [M(II) (L) (L-H)](+) complexes resulted in diagnostic mass spectra, in contrast to the CAD mass spectra of the protonated, deprotonated, and sodium-cationized flavonoid glucosides. Five common sites of glycosylation could be predicted based on the fragmentation patterns of the flavonoid glucoside/magnesium complexes, while flavonoid glucoside/calcium complexes also were effective for location of the glycosylation site when MS(3) was employed. Cobalt, nickel and copper complexation had only limited success in this application. The metal complexation methods were also applied for characterization of a flavonoid rhamnoside, and the dissociation pathways of the metal complexes indicate that flavonoid rhamnosides have distinctive dissociation features from flavonoid glucosides. PMID- 15337510 TI - Mass spectral fragmentation reactions of angiotensin-converting enzyme (ACE) inhibitors. AB - A variety of mass spectrometric techniques have been employed in the study of a series of structurally similar compounds used in the treatment of hypertension. The compounds, known collectively as angiotensin-converting enzyme (ACE) inhibitors, all share the amino acid residue proline or some variant thereof, as a common structural element. The gas phase fragmentation behavior of these compounds has been explored systematically using various instruments and techniques. An interesting dissociation process (rearrangement) unique to one of the compounds, lisinopril, has been investigated using isotopic labeling experiments and exact mass measurements. The general nature of the process has been probed through both the positive and negative ion analyses of fourteen related compounds exhibiting structural homology. PMID- 15337511 TI - Temperature-programmed pyrolysis hyphenated with metastable atom bombardment ionization mass spectrometry (TPPy/MAB-MS) for the identification of additives in polymers. AB - Thermoanalytical techniques are currently used for the analysis of additives contained in polymers that cannot be easily dissolved, extracted, or hydrolyzed. With these techniques, the polymers are heated to liberate the additives trapped in the polymer matrix. If the polymer is heated slowly, up to its thermal degradation, the technique is called temperature-programmed pyrolysis (TPPy). For TPPy experiments, mass spectrometry is generally used as the detection method. The ionization sources commonly used in mass spectrometry, such as CI and EI, can cause fragmentation during the ionization process. Fragmentation decreases the sensitivity of the molecular ions and increases the risks of interferences with the compounds coming from the matrix. An energy-tunable ionization technique, called metastable atom bombardment (MAB), is proposed for TPPy/MS experiments. With this ionization source, the energy of ionization depends on the metastable gas used. With low-energy metastable gases such as Xe or N(2), fragmentation is reduced compared to CI, whereas with medium-energy metastable gases such as Ar or Kr, the fragmentation is similar to that observed with CI. TPPy/MAB-MS was performed on an unknown polyurethane-based car paint. The detection of molecular ions and characteristic fragments with MAB(N(2)) led to the identification of two light stabilizers: Bis(1,2,2,6,6-pentamethyl-4-piperidinyl)sebacate (BPPS) and 2 (2H-benzotriazol-2-yl)-4,6- di-tert-pentylphenol (PTPP). Using MAB(Ar) to simulate CI, the molecular ion and one of the two characteristic fragments of BPPS were not detected, thus confirming the advantage of using MAB(N(2)) ionization for TPPy/MS experiments. PMID- 15337512 TI - Resolution equations for high-field ion mobility. AB - An extension of current mobility resolution equations as they apply to high-field ion mobility spectrometry is presented. The new resolution expression is applied to arrival time distributions for ions having a large range of ion mobilities and mass-to-charge ratios (m/z). The results indicate that the new equation can be utilized to predict the mobility resolution over a broader range of applied electric fields than previous ion mobility resolution expressions. PMID- 15337513 TI - Interpretation of alkyl diselenide and selenosulfenate mass spectra. AB - The mass spectral fragmentation of aliphatic diselenides and selenosulfenates is analyzed to gain a better understanding of the behavior of these species. The main fragmentation pathways of these species include the fragmentation along the Se?C bond, fragmentation along the Se?Se or Se?S bonds and intra-molecular rearrangements. In general, negative ionization favors the fragmentation along the Se?Se or Se?S bonds while positive ionization leads to stable molecular ions. Density functional theory calculations of bond dissociation energies and molecular orbital analysis was undertaken to explain the observed trends in molecular fragmentation. Besides the analysis of molecular fragmentation, a phenomenon of molecular association in negative electron impact and positive chemical ionization conditions was observed and investigated using a high resolution time-of-flight mass spectrometer. Molecular association that occurs during the ionization of species includes the formation of symmetrical diselenides from asymmetrical selenosulfenates and formation of alkylseleno adducts from the corresponding diselenides. For species which is hard to resolve by mass analysis, such as isobars of CHSe, CH(2)Se, and CH(3)Se, the isotope pattern superimposition procedure was applied to define the overlapping clusters. PMID- 15337514 TI - Disposable chromatography for a high-throughput nano-ESI/MS and nano-ESI/MS-MS platform. AB - High-throughput proteomics has typically relied on protein identification based on MALDI-MS peptide maps of proteolytic digests of 2D-gel-separated proteins. This technique, however, requires significant sequence coverage in order to achieve a high level of confidence in the identification. Tandem MS data have the advantage of requiring fewer peptides (2) for high confidence identification, assuming adequate MS/MS sequence coverage. MALDI-MS/MS techniques are becoming available, but can still be problematic because of the difficulty of inducing fragment ions of a singly charged parent ion. Electrospray ionization, however, has the advantage of generating multiply charged species that are more readily fragmented during MS/MS analysis. Two electrospray/tandem mass spectrometry-based approaches, nanovial-ESI-MS/MS and LC-MS/MS, are used for high throughput proteomics, but much less often than MALDI-MS and peptide mass fingerprinting. Nanovial introduction entails extensive manual manipulation and often shows significant chemical background from the in-gel digest. LC-MS has the advantages that the chemical background can be removed prior to analysis and the analytes are concentrated during the separation, resulting in more abundant analyte signals. On the other hand, LC-MS can often be time intensive. Here, we report the incorporation of on-line sample clean-up and analyte concentration with a high-throughput, chip-based, robotic nano-ESI-MS platform for proteomics studies. PMID- 15337515 TI - Nanoflow LC/IMS-MS and LC/IMS-CID/MS of protein mixtures. AB - A simple ion trap/ion mobility/time-of-flight (TOF) mass spectrometer has been coupled with nanoflow liquid chromatography to examine the feasibility of analyzing mixtures of intact proteins. In this approach proteins are separated using reversed-phase chromatography. As components elute from the column, they are electrosprayed into the gas phase and separated again in a drift tube prior to being dispersed and analyzed in a TOF mass spectrometer. The mobilities of ions through a buffer gas depend upon their collision cross sections and charge states; separation based on these gas-phase parameters provides a new means of simplifying mass spectra and characterizing mixtures. Additionally it is possible to induce dissociation at the exit of the drift tube and examine the fragmentation patterns of specific protein ion charge states and conformations. The approach is demonstrated by examining a simple three-component mixture containing ubiquitin, cytochrome c, and myoglobin and several larger prepared protein mixtures. The potential of this approach for use in proteomic applications is considered. PMID- 15337516 TI - Comparative stability determination of oligonucleotide duplexes in gas and solution phase. AB - The relative stability of RNA duplexes were determined in both solution and gas phase. Solution stability as determined by a spectrophotometric method indicated that the Watson-Crick duplexes were more stable than duplexes containing one GA mismatch or two tandem GA mismatches. Gas phase stability was determined using ESI-MS through variation of the collision energy in an ion trap. Stability curves similar to the melting curves obtained in solution were observed. The relative stability in gas phase differed, however, from that in solution. The duplexes with two tandem GA mismatches were found to be more stable than the Watson-Crick and single GA mismatch duplexes. The different trends observed in solution versus gas phase can be attributed to the primary means of interaction. In solution, stacking is expected to be the dominant interaction mode. In the gas phase, hydrogen bonding is expected to be the dominant interaction mode. Duplexes with tandem GA mismatches have the potential to undergo additional hydrogen bonding relative to the other duplexes which could account for their increased stability in the gas phase. PMID- 15337517 TI - Chiral enrichment of serine via formation, dissociation, and soft-landing of octameric cluster ions. AB - Chiral enrichment of serine is achieved in experiments that involve formation of serine octamers starting from non-racemic serine solutions. Serine octamers were generated by means of electrospray and sonic spray ionization of aqueous solutions of d(3)-L-serine (108 Da) and D-serine (105 Da) having different molar ratios of enantiomers. A cyclic process involving the formation of chirally enriched octameric cluster ions and their dissociation, viz. Ser(1) --> Ser(8) - > Ser(1), allows serine monomers to be regenerated with increased enantiomeric excess as shown in two types of experiments: (1) Chiral enrichment in serine was observed in MS/MS/MS experiments in a quadrupole ion trap in which the entire distribution of serine octamers formed from non-racemic solutions was isolated, collisionally activated, and fragmented. Monomeric serine was regenerated with increased enantiomeric excess upon dissociation of octamers when compared with the enantiomeric composition of the original solution. (2) Chiral enrichment was observed in the products of soft-landing of mass-selected protonated serine octamers. These ions were generated by means of electrospray or sonic spray ionization, mass selected, and collected on a gold surface using ion soft landing. Chiral enrichment of the soft-landed serine was established by redissolving the recovered material and comparing the intensities of protonated molecular ions of d(3)-L-serine and D-serine after APCI-MS analysis. Both of these experiments showed comparable results, suggesting that formation of serine octamers depends only on the enantiomeric composition of the serine solution and that the magnitude of the chiral preference is intrinsic to octamers formed from solutions of given chiral composition. PMID- 15337518 TI - Regiospecificity in deuterium labeling determined by mass spectrometry. AB - Several mass spectrometry methods were explored to determine the regiospecificity of deuterium substitutions in hydrocarbon mixtures. The case investigated in this work was that of ethane mixtures obtained by catalytic HD exchange between either C(2)H(6) and D(2) or C(2)D(6) and H(2) over platinum surfaces. A total of ten isotopologs are possible, and were indeed detected in all cases. Deconvolution of low-resolution mass spectra was found sufficient to determine the composition of the gas mixtures in terms of the total number of deuterium substitutions, but not to identify symmetric versus asymmetric substitutions in the C(2)D(2)H(4), C(2)D(3)H(3), and C(2)D(4)H(2) products. High-resolution mass spectrometry allowed the separation of the intensities due to C(2)X(4)(+) fragments from those from molecular C(2)X(6)(+) signals (X = H or D), and with that for a more accurate determination of the composition of the mixtures. Relative probabilities were determined for the symmetric versus asymmetric removal of X(2) from C(2)X(6)(+) ions and for isotope scrambling in the mass spectrometer, and with that information fairly good cracking patterns were then calculated for the C(2)X(4)(+) fragments produced by each individual pure C(2)X(6) isotopologue. However, total deconvolution of all ten components in the ethane mixtures obtained by HD exchange catalysis was beyond the experimental accuracy of the measurements. Tandem mass spectrometry/collision-induced decomposition mass spectrometry (MS/CID-MS) proved more useful for this task. In particular, it was possible to determine the proportion of symmetric versus asymmetric double HD exchange in samples for which the total ethane-d(2) (in the case of C(2)H(6) + D(2)) or ethane-d(4) (with C(2)D(6) + H(2)) amounted to only approximately 3% on the ethane mix. A comparison with other analytical methods, NMR in particular, is provided. PMID- 15337519 TI - Results from a bench marking survey on cassette dosing practices in the pharmaceutical industry. PMID- 15337520 TI - Chemokine receptor CCR5: insights into structure, function, and regulation. AB - CC chemokine receptor 5 (CCR5) is a seven-transmembrane, G protein-coupled receptor (GPCR) which regulates trafficking and effector functions of memory/effector T-lymphocytes, macrophages, and immature dendritic cells. It also serves as the main coreceptor for the entry of R5 strains of human immunodeficiency virus (HIV-1, HIV-2). Chemokine binding to CCR5 leads to cellular activation through pertussis toxin-sensitive heterotrimeric G proteins as well as G protein-independent signalling pathways. Like many other GPCR, CCR5 is regulated by agonist-dependent processes which involve G protein coupled receptor kinase (GRK)-dependent phosphorylation, beta-arrestin-mediated desensitization and internalization. This review discusses recent advances in the elucidation of the structure and function of CCR5, as well as the complex mechanisms that regulate CCR5 signalling and cell surface expression. PMID- 15337521 TI - What turns CREB on? AB - The transactivation domain of the cAMP response element-binding protein (CREB) consists of two major domains. The glutamine-rich Q2 domain, which interacts with the general transcription factor TAFII130/135, is sufficient for the recruitment of a functional RNA polymerase II complex and allows basal transcriptional activity. The kinase-inducible domain, however, mediates signal-induced activation of CREB-mediated transcription. It is generally believed that recruitment of the coactivators CREB-binding protein (CBP) and p300 after signal induced phosphorylation of this domain at serine-133 strongly enhances CREB dependent transcription. Transcriptional activity of CREB can also be potentiated by phosphoserine-133-independent mechanisms, and not all stimuli that provoke phosphorylation of serine-133 stimulate CREB-dependent transcription. This review presents an overview of the diversity of stimuli that induce CREB phosphorylation at Ser-133, focuses on phosphoserine-133-dependent and -independent mechanisms that affect CREB-mediated transcription, and discusses different models that may explain the discrepancy between CREB Ser-133 phosphorylation and activation of CREB-mediated transcription. PMID- 15337522 TI - Adenylate cyclase regulation via proteasome-mediated modulation of Galphas levels. AB - The adenylate cyclase (AC)/cyclic AMP (cAMP)/cAMP-dependent protein kinase pathway controls many biological phenomena. The ubiquitin/proteasome system, controlling the levels of many proteins, modulates important cellular processes such as cell cycle and cell growth. Here we describe a novel mechanism for AC regulation by proteasome pathway. Pharmacological inhibition of proteasome function in human osteosarcoma U2OS cells results in up-regulation of AC activity, increase of levels of alpha subunit of heterotrimeric stimulatory GTP binding proteins (alphas) and, remarkably, also in preventing of beta-adrenergic receptor-mediated down-regulation of alphas protein levels. Accumulation of alphas protein is also accompanied by the appearance of polyubiquitinated alphas species. Our results: (1) identify alphas protein as a novel proteasome substrate in mammalian cells; (2) indicate that proteasome might play a physiological role in controlling AC/cAMP mediated pathways by modulating the levels of Galphas protein; (3) suggest a role for the proteasome also in controlling alphas mediated signaling pathways other than those affecting AC complex. PMID- 15337523 TI - Interleukin-2 (IL-2) receptor-betagamma signalling is activated by c-Kit in the absence of IL-2, or by exogenous IL-2 via JAK3/STAT5 in human papillomavirus associated cervical cancer. AB - Activation of the interleukin-2 receptor (IL-2R) induces signalling cascades promoting T cell proliferation. However, signal transduction pathways triggered in IL-2R-expressing solid tumours are unknown. This report shows that human papillomavirus (HPV)-associated cervical cancer cells express an IL-2R composed of beta and gamma chains (IL-2Rbetagamma), and that IL-2-mediated activation increases the phosphorylation of JAK3 and STAT5, stimulating cell proliferation. Interestingly, endogenous IL-2 is not produced by these cells, suggesting the activation of IL-2Rbetagamma by an alternative mechanism. Accordingly, we found that Stem Cell Factor (SCF)-activated c-Kit induces phosphorylation of the IL 2Rbeta chain in the absence of IL-2. Moreover, inhibition of IL-2Rbeta phosphorylation by blocking c-Kit tyrosine kinase activity abolishes both, IL-2 and SCF-mediated proliferation. Thus, these results demonstrate that IL-2 triggers a JAK3/STAT5 cascade in HPV-associated cervical cancer cells expressing IL-2Rbetagamma, and that this receptor can be alternatively activated by SCF activated c-Kit in the absence of IL-2. PMID- 15337524 TI - Protein tyrosine kinase Syk modulates EGFR signalling in human mammary epithelial cells. AB - Signalling through protein tyrosine kinases (PTKs) is critical in the regulation of important cellular processes and its deregulation is associated with pathophysiological disorders such as cancer. We investigated the function of the PTK spleen tyrosine kinase (Syk) in the regulation of growth factor signalling pathways in human mammary epithelial cells. Our results show that downregulation of endogenous Syk expression enhances the ligand-induced activity of the epidermal growth factor receptor (EGFR) but not that of the closely related human epidermal growth factor receptor 2 (HER2) and human epidermal growth factor receptor 3 (HER3) receptors. Moreover, Syk function interfered with EGFR-mediated cell responses such as proliferation and survival of mammary epithelial cells. A mechanistic link between Syk and EGFR is further supported by the colocalisation of the two PTKs in membrane fractions as well as the regulatory feedback effects of the EGFR kinase on Syk activity. Our findings demonstrate that Syk acts a negative control element of EGFR signalling. PMID- 15337525 TI - Nuclear diacylglycerol kinase-theta is activated in response to nerve growth factor stimulation of PC12 cells. AB - Previous evidence from independent laboratories has shown that the nucleus contains diacylglycerol kinase (DGK) isoforms, i.e., the enzymes, which yield phosphatidic acid from diacylglycerol, thus terminating protein kinase C-mediated signaling events. A DGK isoform, which resides in the nucleus of PC12 cells, is DGK-theta. Here, we show that nerve growth factor (NGF) treatment of serum starved PC12 cells results in the stimulation of both a cytoplasmic and a nuclear DGK activity. However, time course analysis shows that cytoplasmic DGK activity peaked earlier than its nuclear counterpart. While nuclear DGK activity was dramatically down-regulated by a monoclonal antibody known for selectively inhibiting DGK-theta, cytoplasmic DGK activity was not. Moreover, nuclear DGK activity was stimulated by phosphatidylserine, an anionic phospholipid that had no effect on cytoplasmic DGK activity. Upon NGF stimulation, the amount and the activity of DGK-theta, which was bound to the insoluble nuclear matrix fraction, substantially increased. Epidermal growth factor up-regulated a nuclear DGK activity insensitive to anti-DGK-theta monoclonal antibody. Overall, our findings identify nuclear DGK-theta as a down-stream target of NGF signaling in PC12 cells. PMID- 15337526 TI - Role of focal adhesion formation in migration and morphogenesis of endothelial cells. AB - Cell motility and morphogenesis are regulated by a balance between formation and disassembly of stress fibers and focal adhesions. To understand the mechanisms underlying these cellular responses in angiogenesis, we studied the Rho family protein-driven pathways in FGF-2-induced chemotaxis and capillary morphogenesis of murine brain capillary endothelial cell line, IBE cells. Cells seeded onto fibronectin-coated surface migrated toward FGF-2. Expression of dominant negative Rho A (DNRho) or kinase-dead p21-activated kinase 1 (KDPAK1), or treatment with Y27632 inhibited chemotaxis in association with the lack of FGF-2-induced decrease in focal adhesions. On Matrigel, DNRho and Y27632 induced FGF-2 independent capillary morphogenesis despite loss of stress fiber formation. KDPAK1 cells formed stress fibers and showed capillary morphogenesis in response to FGF-2. Increase in focal adhesions was closely associated with capillary morphogenesis. Our results suggest that formation or disassembly of focal adhesions seems to determine the motility or morphogenesis of endothelial cells. PMID- 15337527 TI - Ca2+ release via ryanodine receptors and Ca2+ entry: major mechanisms in NAADP mediated Ca2+ signaling in T-lymphocytes. AB - Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca2+ mobilizing nucleotide essentially involved in T cell activation. Using combined microinjection and single cell calcium imaging, we demonstrate that co-injection of NAADP and the D-myo-inositol 1,4,5-trisphosphate antagonist heparin did not inhibit Ca2+ mobilization. In contrast, co-injection of the ryanodine receptor antagonist ruthenium red efficiently blocked NAADP induced Ca2+ signalling. This pharmacological approach was confirmed using T cell clones stably transfected with plasmids expressing antisense mRNA targeted specifically against ryanodine receptors. NAADP induced Ca2+ signaling was strongly reduced in these clones. In addition, inhibition of Ca2+ entry by SK&F 96365 resulted in a dramatically decreased Ca2+ signal upon NAADP injection. Gd3+, a known blocker of Ca2+ release activated Ca2+ entry, only partially inhibited NAADP mediated Ca2+ signaling. These data indicate that in T cells (i) ryanodine receptor are the major intracellular Ca2+ release channels involved in NAADP induced Ca2+ signals, and that (ii) such Ca2+ release events are largely amplified by Ca2+ entry. PMID- 15337528 TI - c-Cbl binds to tyrosine-phosphorylated neurotrophin receptor p75 and induces its ubiquitination. AB - The p75 neurotrophin receptor (p75NTR) has dual functions in cell survival and cell death but its intracellular signalling pathways are not understood. Here we describe that in rat brain and in pervanadate-stimulated PCNA and HEK293 cells p75NTR is phosphorylated at a single tyrosine residue within the cytosolic C terminus. Phosphorylated tyrosine 308 constitutes a binding site for the ubiquitin ligase c-Cbl. This interaction is a prerequisite for ubiquitination of p75NTR. Our data suggest a c-Cbl-dependent ubiquitination of p75NTR involved in the regulation of p75NTR signalling. PMID- 15337529 TI - Src tyrosine kinase regulates insulin-induced activation of protein kinase C (PKC) delta in skeletal muscle. AB - Insulin stimulation of skeletal muscle results in rapid activation of protein kinase Cdelta (PKCdelta), which is associated with its tyrosine phosphorylation and physical association with insulin receptor (IR). The mechanisms underlying tyrosine phosphorylation of PKCdelta have not been determined. In this study, we investigated the possibility that the Src family of nonreceptor tyrosine kinases may be involved upstream insulin signaling. Studies were done on differentiated rat skeletal myotubes in primary culture. Insulin caused an immediate stimulation of Src and induced its physical association with both IR and PKCdelta. Inhibition of Src by treatment with the Src family inhibitor PP2 reduced insulin-stimulated Src-PKCdelta association, PKCdelta tyrosine phosphorylation and PKCdelta activation. PP2 inhibition of Src also decreased insulin-induced IR tyrosine phosphorylation, IR-PKCdelta association and association of Src with both PKCdelta and IR. Finally, inhibition of Src decreased insulin-induced glucose uptake. We conclude that insulin activates Src tyrosine kinase, which regulates PKCdelta activity. Thus, Src tyrosine kinase may play an important role in insulin-induced tyrosine phosphorylation of both IR and PKCdelta. Moreover, both Src and PKCdelta appear to be involved in IR activation and subsequent downstream signaling. PMID- 15337530 TI - Drosophila PI3 kinase and Akt involved in insulin-stimulated proliferation and ERK pathway activation in Schneider cells. AB - We have characterized the role of Drosophila PI3K and AKT in ERK pathway activation involving insulin-induced proliferation using Drosophila Schneider cells. After insulin treatment, dPI3K and dAKT activities were both increased along with activation of the dERK pathway components dMEK and dERK. The insulin induced activations of dERK and dAKT were blocked by LY294002, dPTEN, and by an AKT inhibitor, indicating involvement of dPI3K and dAKT in the insulin-induced dERK and dAKT activations. Proliferation and the G1 to S phase cell cycle progression due to insulin were also blocked by PI3K and AKT inhibitors, indicating that the Drosophila PI3K-AKT pathway involves insulin-mediated cell proliferation. The insulin-stimulated size increase was blocked by both LY294002 and AKT inhibitor, not by U0126, indicating that insulin-mediated size control by dPI3K and dAKT occurs independently of the ERK pathway. This study indicates that dPI3K and dAKT are involved in insulin-induced ERK pathway activation leading to proliferation in Drosophila Schneider cells. PMID- 15337531 TI - Ras inhibition leads to transcriptional activation of p53 and down-regulation of Mdm2: two mechanisms that cooperatively increase p53 function in colon cancer cells. AB - Activated Ras, operating through the Raf/MEK/ERK pathway, is known to regulate transcription of both Mdm2 and its inhibitor p19ARF, resulting in opposing effects on the tumor suppressor protein p53. We show here that a decrease in Ras in SW480 cells induced either by the Ras inhibitor farnesylthiosalicylic acid (FTS) or by K-Ras antisense oligonucleotides, resulted in a similar increase in p53 protein. The increase in p53 was accompanied by an increase in p21(waf1/cip1) mRNA transcripts and protein. Consistent with the Ras/Raf/MEK/ERK-mediated control of Mdm2, treatment of SW480 cells with the Ras inhibitor FTS caused a marked (80%) decrease in Mdm2, which itself would account for the increase in p53. However, FTS also caused a 1.6-fold increase in p53 mRNA, indicative of a Ras-dependent mechanism that regulates p53 transcription. Thus, oncogenic Ras appears to attenuate p53 in SW480 cells by two independent regulatory mechanisms, the one leading to increased Mdm2-dependent p53 degradation and the other leading to a decrease in p53 transcription. PMID- 15337532 TI - Are radiation oncologists serious about systemic radiation therapy and if not, should we be? PMID- 15337533 TI - High-tech will improve radiotherapy of NSCLC: a hypothesis waiting to be validated. PMID- 15337534 TI - A historical perspective of the radiation oncology workforce and ongoing initiatives to impact recruitment and retention. AB - The field of radiation oncology has evolved into an exceptionally technologically driven and multi-disciplinary discipline over the last two decades. This specialty of medicine is one that requires not only the command of highly complex modalities but also the assembly of a competent and expertly skilled team of medical professionals. Although the profession has grown tremendously in the past years, the workforce has not been able to meet the demands of the practice. A significant shortage of radiation therapists, dosimetrists, and oncology nurses exists in the United States today and will almost certainly increase in severity over the next several years. A similar crisis has been seen in several other countries-most notably Canada and Australia and has contributed to prolonged delays in cancer treatment for many patients. PMID- 15337535 TI - Report of a multicenter Canadian phase III randomized trial of 3 months vs. 8 months neoadjuvant androgen deprivation before standard-dose radiotherapy for clinically localized prostate cancer. AB - PURPOSE: To evaluate the effect of 3 months vs. 8 months of neoadjuvant hormonal therapy before conventional dose radiotherapy (RT) on disease-free survival using prostate-specific antigen PSA and biopsies as end points for clinically localized prostate cancer. METHODS AND MATERIALS: Between February 1995 and June 2001, 378 men were randomized to either 3 or 8 months of flutamide and goserelin before conventional-dose RT (66 Gy) at four participating centers. The median patient age was 72 years (range, 50-84 years). The stage distribution was 17% T1c, 35% T2a, 34% T2b-T2c, 13% T3-T4. The Gleason score (GS) was < or =6 in 51%, 7 in 38%, and 8-10 in 11%. The median baseline PSA level was 9.7 ng/mL (range, 1.3-189 ng/mL). Of the 378 men, 26% were low risk (Stage T1c-T2a, GS < or =6, PSA <10 ng/mL), 43% were intermediate risk (Stage T2b or GS 7 or PSA 10-20 ng/mL), and 31% were high risk (Stage T3 or GS 8-10 or PSA >20 ng/mL). The two arms were balanced in terms of age, GS, T stage, risk group, and presenting PSA level. The median follow-up was 44 months (range, 10-84 months), and 361 patients were available for evaluation. RESULTS: The 8-month arm achieved a lower PSA level before starting RT (0.37 vs. 0.74 ng/mL, p < or =0.001) and had a greater downsizing of the prostate (mean volume 26.6 cm(3) vs. 30.5 cm(3), p < or =0.001). However, the actuarial freedom from failure rate (biochemical by American Society for Therapeutic Radiology and Oncology definition, local or distant) for the 3-month vs. 8-month arms at 3 years was 66% vs. 68% and by 5 years was 61% vs. 62%, respectively (p = 0.36). No statistically significant difference was noted in the types of failure between the two arms (crude final status): biochemical, 22.2% vs. 22.3%; local, 10.2% vs. 6.5%; and distant, 3.4% vs. 4.4% (p = 0.61). Two-year post-RT biopsies were done in 57% (n = 205). Negative biopsies were obtained in 68% of the 3-month and 77% of the 8-month patients; 18% and 14% had indeterminate biopsies and 14% and 9% were positive for residual cancer (p = 0.34) in the two arms, respectively. The median PSA level for nonfailing patients was 0.50 ng/mL in both the 3-months and 8-month arms. A suggestion of improvement was found in the 8-month arm for disease-free survival at 5 years for high-risk patients (39% vs. 52%) but did not achieve statistical significance. CONCLUSION: A longer period of neoadjuvant hormonal therapy before standard-dose RT does not appear to confer a benefit in terms of disease-free survival or to alter failure patterns. Failure was delayed in the 8-month arm, but this advantage was lost by 5 years of follow-up. A suggestion of benefit was noted with a longer period of hormonal therapy for high-risk patients. PMID- 15337536 TI - Association of percent positive prostate biopsies and perineural invasion with biochemical outcome after external beam radiotherapy for localized prostate cancer. AB - PURPOSE: Few studies have evaluated the significance of the percentage of positive biopsies (PPB) and perineural invasion (PNI) for patients treated with external beam radiotherapy (EBRT) for localized prostate cancer. Our goal was to investigate the value of these factors in predicting biochemical control (bNED) after EBRT. METHODS AND MATERIALS: The study cohort consisted of 331 patients who received EBRT between 1993 and 1999 for clinically localized prostate cancer. The median follow-up was 4.4 years (range, 3 months to 9.6 years). The distribution by clinical T stage was as follows: T1 in 55 (17%), T2a in 94 (28%), T2b in 76 (23%), T2c in 74 (22%), T3a in 27 (8%), and T3b in 5 (2%). The pretreatment prostate-specific antigen (iPSA) level was < or =10 ng/mL in 224 patients, 10.1 20 ng/mL in 72 patients, and >20 ng/mL in 35 patients. The biopsy Gleason score was < or =6 in 216 patients and > or =7 in 115 patients. On the basis of the pathology report, the PPB was calculated for 239 patients and was < or =33% in 109, 34-66% in 72, and > or =67% in 58 patients. PNI was present in 30 patients. The median dose of EBRT was 68.4 Gy (range, 64-71 Gy). Patients were categorized into three risk groups: 142 patients were low risk (T1-T2, iPSA < or =10 ng/mL, and Gleason score < or =6), 137 were intermediate risk (increase in the value of one of the risk factors); and 52 patients were high risk (increase in value of two or more of the risk factors). Biochemical failure was defined as three consecutive rises in the PSA level. RESULTS: The 5-year bNED rate for the entire cohort was 62%. The 5-year bNED rate for the low-, intermediate, and high-risk group was 79%, 51%, and 47%, respectively (p <0.0001). On univariate analysis (log-rank test), clinical stage (p = 0.0073), grade (p <0.0001), iPSA (p = 0.0043), risk group (p <0.0001), PPB (p = 0.0193), and presence of PNI (p = 0.0137) correlated with bNED. For T1-T2a, T2b-T2c, and T3 patients, the 5-year bNED rate was 71%, 59%, and 40%, respectively. The 5-year bNED rate was 68% for those with an iPSA level of < or =10 ng/mL and 49% for those with an iPSA level of >10 ng/mL. For patients with PPB < or =33%, 34-66%, and > or =67%, the 5-year bNED rate was 75%, 67%, and 51%, respectively. Within the intermediate-risk group, the PPB was significantly associated with the bNED rate: 67%, 52%, and 30% for those with PPB < or =33%, 34-66%, and > or =67%, respectively (p = 0.0046). This association was not seen in the low- or high-risk group. The 5-year bNED rate was 64% for patients without PNI and 48% for those with PNI. On multivariate analysis (Cox proportional hazards model), the statistically significant predictive factors for bNED were risk group (p = 0.0032) and PPB (p = 0.044). Using the chi-square test, statistically significant associations between T stage, PSA level, Gleason score, and risk group with PPB were found; PNI was significantly associated with T stage and PSA level only. CONCLUSION: Our results showed that PPB and PNI have a statistically significant impact on the bNED rate in patients treated with conventional dose of EBRT (< or =71 Gy). Within the intermediate-risk group, the PPB was predictive of bNED, suggesting that patients with < or =33% PPB had a statistically significant better treatment outcome compared with those with a greater PPB. PNI was not significant for bNED in multivariate analysis. The effects of these two prognostic factors in patients who have been treated with higher doses of RT (> or =75.6 Gy) should be studied. PMID- 15337537 TI - Measurements of intrafraction motion and interfraction and intrafraction rotation of prostate by three-dimensional analysis of daily portal imaging with radiopaque markers. AB - PURPOSE: To measure the interfraction and intrafraction motion of the prostate during the course of external beam radiotherapy using a video electronic portal imaging device and three-dimensional analysis. METHODS AND MATERIALS: Eighteen patients underwent implantation with two or three gold markers in the prostate before five-angle/11-field conformal radiotherapy. Using CT data as the positional reference, multiple daily sets of portal images, and a three dimensional reconstruction algorithm, intrafraction translations, as well as interfraction and intrafraction rotations, were analyzed along the three principal axes (left-right [LR], superoinferior [SI], and AP). The overall mean values and standard deviations (SDs), along with random and systematic SDs, were computed for these translations and rotations. RESULTS: For 282 intrafraction translational displacements, the random SD was 0.8 mm (systematic SD, 0.2) in the LR, 1.0 mm (systematic SD, 0.4) in the SI, and 1.4 mm (systematic SD, 0.7) in the AP axes. The analysis of 348 interfraction rotations revealed random SDs of 6.1 degrees (systematic SD, 5.6 degrees ) around the LR axis, 2.8 degrees (systematic SD, 2.4 degrees ) around the SI axis, and 2.0 degrees (systematic SD, 2.2 degrees ) around the AP axis. The intrafraction rotational motion observed during 44 fractions had a random SD of 1.8 degrees (systematic SD, 1.0 degrees ) around the LR, 1.1 degrees (systematic SD, 0.8 degrees ) around the SI, and 0.6 degrees (systematic SD, 0.3 degrees ) around the AP axis. CONCLUSION: The interfraction rotations observed were more important than those reported in previous studies. Intrafraction motion was generally smaller in magnitude than interfraction motion. However, the intrafraction rotations and translations of the prostate should be taken into account when designing planning target volume margins because their magnitudes are not negligible. PMID- 15337538 TI - 125I reimplantation in patients with poor initial dosimetry after prostate brachytherapy. AB - PURPOSE: In this case report, we describe 7 patients with suboptimal dosimetry after (125)I prostate brachytherapy who underwent a second implant procedure to improve the dosimetric coverage of the prostate. METHODS AND MATERIALS: Seven patients underwent second (125)I implants for suboptimal dosimetry after their initial implant for prostate cancer. The pretreatment characteristics (clinical stage, Gleason score, initial prostate-specific antigen level, location of positive cores, International Prostate Symptom Score, potency, use of androgen suppression, initial implant planning characteristics) were noted. The "Day 30" CT-based dosimetry parameters after the first implant and "Day 0" CT-based dosimetry after the reimplant were recorded (volumes of prostate and rectum covered by 100% and 150% of the dose and dose covering 90% of the prostate volume). The toxicity of the second procedure, International Prostate Symptom Scores before and after reimplantation, the clinical course, and prostate specific antigen outcomes after reimplant were examined. We described our reimplant planning and intraoperative procedure. RESULTS: In all 7 patients, we were able to achieve very favorable dosimetry after the second procedure. The acute toxicity of the reimplant procedure was reasonably low, and the short-term prostate-specific antigen outcome has been favorable. CONCLUSION: It is possible to add more seeds safely to the dosimetrically cool area after the initial brachytherapy procedure and achieve excellent postimplant dosimetry with acceptable acute toxicity. The ultimate benefits and long-term toxicity of reimplantation are unknown. PMID- 15337539 TI - Health-related quality of life in men after treatment of localized prostate cancer with external beam radiotherapy combined with (192)ir brachytherapy: a prospective study of 93 cases using the EORTC questionnaires QLQ-C30 and QLQ PR25. AB - PURPOSE: To describe prospectively the long-term health-related quality of life (HRQOL) and treatment-related symptoms in patients with localized prostate cancer treated with neoadjuvant androgen deprivation therapy and radical radiotherapy (RT), including external beam RT and iridium high-dose-rate brachytherapy, and to compare them with age-matched normative data. METHODS AND MATERIALS: A total of 93 patients with T1-T3a tumors consecutively treated with definitive RT at our institution completed the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (QLQ)-C30 and QLQ-prostate specific 25-item (PR25) module twice at an 18-month interval 0-18 months after RT. Subgroups were analyzed regarding acute and late effects on symptoms and quality of life. RESULTS: The main analysis included 80 patients who were disease free at the final assessment. The levels of HRQOL were generally high, did not change over time, and were comparable to the normative data. Symptom development (urinary, bowel, and sexual) correlated well with the known acute and late effects of radical RT and neoadjuvant androgen deprivation therapy. CONCLUSION: Combining external beam RT and HDR brachytherapy when treating prostate cancer did not appear to impair HRQOL and was comparable to that of other brachytherapy methods. The negative contribution from late neoadjuvant androgen deprivation therapy on symptom development seemed to be substantial but mostly transitory. Additional research is needed to determine the long-term HRQOL (3-5 years), and interventional randomized studies are suggested. PMID- 15337540 TI - Clinical radiobiology of stage T2-T3 bladder cancer. AB - PURPOSE: To evaluate the relationship between total radiation dose and overall treatment time (OTT) with the treatment outcome, with adjustment for selected clinical factors, in patients with Stage T2-T3 bladder cancer treated with curative radiotherapy (RT). METHODS AND MATERIALS: The analysis was based on 480 patients with Stage T2-T3 bladder cancer who were treated at the Center of Oncology in Gliwice between 1975 and 1995. The mean total radiation dose was 65.5 Gy, and the mean OTT was 51 days. In 261 patients (54%), planned and unplanned gaps occurred during RT. Four fractionation schedules were used: (1) conventional fractionation (once daily, 1.8-2.5 Gy/fraction); (2) protracted fractionation (pelvic RT, once daily, 1.6-1.7 Gy/fraction, boost RT, once daily, 2.0 Gy/fraction); (3) accelerated hyperfractionated boost (pelvic RT, once daily, 2.0 Gy/fraction; boost RT, twice daily, 1.3-1.4 Gy/fraction); and (4) accelerated hyperfractionation (pelvic and boost RT, twice daily, 1.2-1.5 Gy/fraction). In all fractionation schedules, the total radiation dose was similar (average 65.5 Gy), but the OTT was different (mean 53 days for conventional fractionation, 62 days for protracted fractionation, 45 days for accelerated hyperfractionated boost, and 41 days for accelerated hyperfractionation). A Cox proportional hazard model and maximum likelihood logistic model were used to evaluate the relationship between the treatment-related parameters (total radiation dose, dose per fraction, and OTT) and clinical factors (clinical T stage, hemoglobin level and bladder capacity before RT) and treatment outcome. RESULTS: With a median follow-up of 76 months, the actuarial 5-year local control rate was 47%, and the overall survival rate was 40%. The logistic analysis, which included the total dose, OTT, and T stage, revealed that all of these factors were significantly related to tumor control probability (p = 0.021 for total radiation dose, p = 0.038 for OTT, and p = 0.00068 for T stage). A multivariate Cox model, which included the treatment-related parameters and other clinical factors, revealed that the hemoglobin level and bladder capacity before RT and T-stage were statistically significant factors determining local control and overall survival. The total radiation dose was of borderline statistical significance for overall survival (p = 0.087), and OTT did not reach statistical significance. CONCLUSION: The results of our study showed that the treatment outcome after RT for bladder cancer depends mainly on clinical factors: hemoglobin level and bladder capacity before RT, and clinical T stage. An increase in the total radiation dose seemed to be associated with a better treatment outcome. The effect of the OTT was difficult to define, because it was influenced by other prognostic factors. PMID- 15337541 TI - Breast-conserving therapy for stage I-II breast cancer in elderly women. AB - PURPOSE: To assess breast-conserving therapy results in elderly patients with early-stage breast cancer (clinical Stage I-II). METHODS AND MATERIALS: Between 1979 and 1998, 196 women (200 treated breasts) aged > or =70 years (median age, 72.5 years) were treated with breast-conserving therapy (lumpectomy or quadrantectomy with axillary lymph node dissection and radiotherapy). Pathologic axillary node involvement was found in 51 patients (28%). Two-thirds of patients received tamoxifen, and 16% received chemotherapy. RESULTS: At a median follow-up of 59 months, 3 patients (1.5%) had developed local recurrence and 20 (10.2%) distant metastases. The overall survival rate was 81% and 62% at 5 and 10 years, respectively. The corresponding disease-specific survival rates were 92% and 88%. Axillary nodal involvement was the only statistically significant risk factor for the development of metastases (p = 0.0035). Arm mobility impairment and arm lymphedema each occurred in 5 patients. In another 5 patients, a thromboembolic event occurred during tamoxifen treatment. CONCLUSION: Elderly women tolerate breast-conserving therapy, including radiotherapy, well and have excellent rates of locoregional control and disease-specific survival. PMID- 15337542 TI - Status of postmastectomy radiotherapy in the United States: a patterns of care study. AB - PURPOSE: The Patterns of Care Study performed this first known practice survey to establish a national profile of the delivery of postmastectomy radiotherapy (RT) in operable breast cancer. METHODS AND MATERIALS: A Patterns of Care Study research associate collected data from 55 randomly selected institutions. The survey data included 132 items describing the patient, pathologic features, and treatment course for patients with clinical Stage I, II, and IIIA breast cancer undergoing postmastectomy RT in 1998 and 1999. A multivariate analysis was performed to determine the impact of tumor factors and type of treatment facility on the radiation fields used. RESULTS: A weighted sample size of 13,720 was obtained from a sampling of 405 patient records. The mean tumor size was 3.5 cm, and the mean number of axillary nodal metastases was 4.55. Lymphatic vascular invasion was noted in 34%, microscopic skin or dermal lymphatic invasion in 16%, positive or close margins in 36%, and extracapsular nodal extension in 23%. Radiotherapy included the chest wall in all cases and the regional nodes in 78%. When nodal RT was delivered, it included a supraclavicular field, supplemental axillary field, and/or an internal mammary field in 98%, 46%, and 23% of cases, respectively. Chest wall and supraclavicular RT was delivered in >90% of instances with 6-MV photons to doses between 45 and 50 Gy. More variation was seen in the delivery of the axillary and internal mammary RT. On multivariate analysis, the presence of four or more positive nodes and treatment at a large volume facility were the factors most frequently associated with the use of regional radiation fields. CONCLUSION: This Patterns of Care Study survey has demonstrated that breast cancer patients undergoing postmastectomy RT in 1998 and 1999 had a high proportion of factors associated with an increased risk of locoregional failure. The practice patterns established in this study provide a baseline for comparison with future survey results. PMID- 15337543 TI - Use of pathologic factors to assist in establishing adequacy of excision before radiotherapy in patients treated with breast-conserving therapy. AB - BACKGROUND: We reviewed our experience in patients with early-stage breast carcinoma treated with breast-conserving therapy (BCT) to identify pathologic factors useful in defining the adequacy of excision before radiotherapy (RT). METHODS AND MATERIALS: All slides from 607 consecutively treated cases of Stage I II breast carcinoma were reviewed by one pathologist. Numerous pathologic factors were evaluated for their association with ipsilateral breast failure (IBF). Margin distance was classified as negative, near (<0.50 low-power field), or positive. The amount of carcinoma near the final margin was measured as the width of invasive carcinoma and number of ducts with carcinoma in situ near the margin and divided into three groups: near-least, near-intermediate, and near-greatest amount. The median follow-up was 8.5 years. RESULTS: Patients with negative, near least, near-intermediate, and near-greatest amount of carcinoma near the margin, and positive final margins had a 12-year IBF rate of 9%, 6%, 18%, 24%, and 30%, respectively. On multivariate analysis, only the amount of carcinoma near the margin was independently associated with IBF (p <0.001). To help explain these observations, 441 initial and reexcision specimens were examined. The amount of carcinoma near the initial margin and the invasive carcinoma/specimen maximal dimension ratio were significantly associated with greater amounts of residual carcinoma in adjacent breast parenchyma. CONCLUSION: The amount of carcinoma near the margin, in addition to margin status, appears to be directly related to an increased risk of IBF in patients treated with BCT. Pathologic factors that incorporate the amount of excised breast parenchyma and amount of carcinoma near the margin may be useful to clinicians in deciding whether a patient has undergone adequate excision for BCT. PMID- 15337544 TI - Dosimetric analysis of a simplified intensity modulation technique for prone breast radiotherapy. AB - PURPOSE: Prone-position breast radiotherapy (RT) has been described as an alternative technique to improve dose homogeneity for women with large, pendulous breasts. We report the feasibility and dosimetric analysis of a simplified intensity-modulated RT (IMRT) technique, previously reported for women in the supine treatment position, to plan prone-position RT to the intact breast. METHODS AND MATERIALS: Twenty patients with clinical Stage TisN0-T1bN1 breast cancer undergoing breast-conserving therapy underwent whole breast RT using a prone position technique. The treatment plans were developed using both conventional tangents and a simplified intensity-modulated tangential beam technique based on optimization of the intensity distributions across the breast. The plans were compared with regard to the dose-volume parameters. RESULTS: Dose heterogeneity within the breast planning target volume was significantly greater for the conventional tangent plans. Of 20 patients, 16 (80%) received maximal doses of > or =110% using the conventional tangents vs. only 1 (5%) using the IMRT plan. The isodose level encompassing 5% of the planning target volume was reduced from an average of 110% with conventional tangents to 105% with IMRT. The maximal dose within the planning target volume was reduced from an average of 114% with conventional tangents to 107% with IMRT. The greatest improvement was seen in the patients with the most pendulous breasts. CONCLUSION: An IMRT planning approach is feasible for prone-position breast RT and improves dose homogeneity, particularly in women with larger, pendulous breasts. Additional follow-up is necessary to determine whether the improvements in dose homogeneity impact acute toxicity and cosmetic outcome in this cohort of women who have historically suffered from poor cosmesis after breast-conserving therapy. PMID- 15337545 TI - Regional radiotherapy to axilla and supraclavicular fossa for adjuvant breast treatment: a comparison of four techniques. AB - PURPOSE: The three techniques commonly used to treat the axilla and supraclavicular nodes in adjuvant radiotherapy all have significant disadvantages, including underdosing the deeper nodes, excessively irradiating normal tissues, or producing undesirable hot spots. We assessed whether an anterior field with posterior boost field to the axilla with customized compensation of the anterior beam (APcomp-PAboost) would minimize these drawbacks. METHODS AND MATERIALS: The axillary and supraclavicular nodal volumes, planning target volume (PTV), irradiated volume, and brachial plexus were contoured for 10 patients. The plans for each technique-single anterior field (AP); anterior to posterior parallel pair (AP-PA); anterior field with posterior boost (AP-PAboost); and APcomp-PAboost-were then generated for each patient using CadPlan and compared. RESULTS: The AP plan gave poor PTV coverage in 60% of cases. The AP-PA provided good PTV coverage and minimal hot spots, but resulted in consistent unnecessary RT to the medial posterior neck. The skin and tissue of the medial posterior neck and chest wall (i.e., the tissue overlying the posterior half of the ribs and posterior to the latissimus dorsi muscle, which forms the posterior wall of the axilla) was incidentally included in the radiation fields of the AP-PA and the exit of the AP beam. No nodal tissue is present in this region, and, therefore, this tissue was unnecessarily irradiated to higher doses with the AP-PA technique. The AP-PAboost provided adequate PTV coverage and a limited dose to the medial posterior neck, but produced hot spots in excess of 120% in 90% of cases. The APcomp-PAboost provided good PTV coverage, a limited dose to the medial posterior neck, and hot spots to <120% in all cases. CONCLUSION: In most cases, the APcomp-PAboost technique offered the best compromise, but the AP-PA technique may be preferred if a less intense hot spot is sought. PMID- 15337546 TI - Twice-daily radiotherapy as concurrent boost technique during two chemotherapy cycles in neoadjuvant chemoradiotherapy for resectable esophageal carcinoma: mature results of phase II study. AB - PURPOSE: To determine the toxicities of neoadjuvant chemoradiotherapy using a three-drug regimen (cisplatin, 5-fluorouracil, and paclitaxel) and a conventional radiotherapy (RT) schedule combined with a concurrent boost technique during chemotherapy cycles, and to determine the rate of tumor response, overall survival, and impact of pathologic tumor response on survival. METHODS AND MATERIALS: The eligibility criteria included resectable adenocarcinoma or squamous cell carcinoma (T2-T3N0-N1M0), performance score < or =2, and no significant comorbidities for trimodality therapy. Chemotherapy consisted of two cycles of cisplatin, 5-fluorouracil, and paclitaxel. A concurrent boost technique was used in RT for 2 levels of radiation doses: 58.5 Gy in 34 fractions within 5 weeks to the gross tumor volume and 45 Gy in 25 fractions within 5 weeks to the clinical target volume by administering a boost dose of 13.5 Gy in 9 fractions, 1.5 Gy/fraction, as a second daily fraction for 9 days on Days 1-5 and 29-32 of the chemotherapy cycles. RESULTS: We enrolled 46 patients in the study. The paclitaxel dose was started at 75 mg/m(2) (n = 7) and escalated to 125 mg/m(2) (n = 5), at which point, dose-limiting toxicities occurred. Thereafter, paclitaxel at 100 mg/m(2) was used for an additional 34 patients. Toxicities included Grade 4 neutropenia (22%), febrile neutropenia requiring hospital admission (20%), Grade 3 (48%) and Grade 4 (7%) acute esophagitis, and paclitaxel-associated anaphylaxis (4%). Of the 46 patients, 3 (6.5%) died of treatment-related complications, 1 of pneumonia during induction therapy and 2 of postoperative complications (5% of the 40 patients who underwent resection). The histopathologic tumor response was a pathologic complete response (pT0N0) in 18 (45%) of 40 patients who underwent resection and 18 (39%) of all 46 registered patients. Minimal residual disease (pT1N0) at the primary site was present in 5 (11%) and residual disease in 23 (50%) of all 46 patients. The minimal follow-up for all long-term survivors (n = 16) was 5.5 years. The median survival time was 34 months, and the overall survival rate was 57%, 50%, and 37% at 2, 3, and 5 years, respectively. The 5-year overall survival (56% vs. 24%, p = 0.0214) and disease-free survival (48% vs. 6%) were significantly better statistically for patients with a pathologic complete response and minimal residual disease than for those with residual disease. All long-term survivors beyond 5.5 years without recurrence accrued from patient cohorts with a pathologic complete response or minimal residual disease. CONCLUSION: An incorporation of twice-daily RT as a concurrent boost to the conventional daily RT schedule during chemotherapy cycles is feasible and warrants additional study for radiation dose intensification. Such research would be prudent for both improved long-term survival and organ preservation in esophageal carcinoma. PMID- 15337547 TI - Late-course accelerated hyperfractionated radiotherapy for localized esophageal carcinoma. AB - PURPOSE: To evaluate the long-term survival results and patterns of failure for localized carcinoma of the esophagus receiving late-course accelerated hyperfractionated (LCAF) radiotherapy (RT). METHODS AND MATERIALS: We studied 201 patients with histologically confirmed squamous cell carcinoma of the esophagus who were treated with LCAF RT between August 1994 and January 2000. The design of the radiation fields was based on the diagnosis by computed tomography and barium examination. All patients received conventionally fractionated RT at 1.8 Gy/d, five fractions weekly for the first two-thirds of treatment to a dose of about 41.4 Gy in 23 fractions within 4-5 weeks. This was followed by LCAF RT using reduced fields, 1.5 Gy/fraction twice daily with a 6-h interval between fractions, to a dose of about 27 Gy within 9 days. Thus, the total dose was 68.4 Gy in 41 fractions within 44 days. RESULTS: The incidence of Grade 3-5 acute radiation-induced bronchitis was 4.0% (8 cases), 3.0% (6 cases), and 0%, respectively. The incidence of Grade 3-5 acute radiation-induced esophagitis was 14.9% (30 cases), 0.5% (1 case), and 0%. Ten patients (5%) died of late complications. The 1-year, 3-year, and 5-year overall survival rate was 73%, 34%, and 26%, respectively. The 1-year, 3-year, and 5-year local control rate was 77%, 58%, and 56%, respectively. The main site of first failure was locoregional failure and distant metastasis (including lymph node metastasis from regional recurrence). Of 201 patients, 77 (38.4%) had local disease alone or with distant metastasis as the first failure, and 70 patients (34.9%) had distant metastasis and/or lymph node metastasis alone or with local failure as the first failure. CONCLUSION: The LCAF regimen offers similar local control and survival to standard chemotherapy plus RT, such as was delivered in the Radiation Therapy Oncology Group studies 85-01 and 94-05. PMID- 15337548 TI - Chemoradiation with raltitrexed (Tomudex) in preoperative treatment of stage II III resectable rectal cancer: a phase II study. AB - PURPOSE: To evaluate the impact of preoperative chemoradiation with raltitrexed (Tomudex(1)) on tumor response, sphincter preservation, and toxicity in patients with locally advanced rectal cancer. METHODS AND MATERIALS: Between 1998 and 2002, 54 consecutive patients with Stage T3 or T2N+ resectable rectal carcinoma were treated with preoperative chemoradiation, i.v. bolus of raltitrexed on Days 1, 19, and 38 and concurrent 50 Gy external beam radiotherapy. Surgery was performed 6-8 weeks after the end of chemoradiation. RESULTS: No patients had Grade 4 acute toxicity. Grade 3 acute toxicity occurred in 16.6% of cases and was hematologic in 6 patients and GI in 2. The overall clinical response rate was 88.8%, with a complete response in 5.5%, partial response in 83.3%, and no change in 9.2%. No patient showed disease progression. All patients underwent surgery. Sphincter saving was obtained in 83.3% of patients. No perioperative mortality occurred, and the perioperative morbidity rate was 5.5%. Of 20 resected patients (37%) who were candidates for abdominoperineal resection at diagnosis (anorectal ring distance < or =30 mm), 13 (65%) underwent a sphincter-saving procedure. At pathologic examination, 13 (24%) of 54 patients had a complete pathologic response (pT0) and 10 (18.5%) had rare isolated residual cancer cells (pT, microscopic foci). Overall, 42.5% had major downstaging. The tumor regression grade (TRG), using Mandard's score system, was also applied and was TRG1 in 13 patients, TRG2 in 11, TRG3 in 20, and TRG4 in 10 patients; no patient had TRG5. CONCLUSION: The use of raltitrexed in a neoadjuvant chemoradiation schedule promoted high pathologic tumor downstaging and use of a sphincter-saving procedure. The low toxicity profile supports the rationale to explore raltitrexed combined with other drugs with different biologic targets. PMID- 15337549 TI - Chemoradiation with raltitrexed and oxaliplatin in preoperative treatment of stage II-III resectable rectal cancer: Phase I and II studies. AB - PURPOSE: Two separate studies were conducted, the first to evaluate the maximal tolerated dose and the second the efficacy of raltitrexed plus oxaliplatin in conjunction with preoperative chemoradiation in patients with resectable T3 rectal carcinoma. METHODS AND MATERIALS: A total of 48 patients received radiotherapy (50 Gy) administered to the posterior pelvis 5 d/wk for 5 weeks. Combination raltitrexed (3 mg/m(2)) and oxaliplatin (60 to 130 mg/m(2)) was administered on Days 1, 19, and 38. RESULTS: The recommended dose of oxaliplatin is 130 mg/m(2) (maximal tolerated dose not reached). No patients developed Grade 4 acute toxicity. Grade 3 acute toxicity occurred in 9 patients (18.7%). It was hematologic in 1 patient and GI in 1 patient; 7 patients had an asymptomatic increase of transaminase. Surgery was performed in 47 (98%) of 48 patients. Of the 47 patients, 42 underwent sphincter-saving surgery; in 19, the tumor at diagnosis was located <30 mm from the anorectal ring. Chemoradiation in combination with raltitrexed and oxaliplatin produced high rates of tumor response. The overall tumor downstaging rate was 73% for T and N stages. A complete pathologic tumor response (pT0) or microscopic tumor foci (pTmic) was observed in 28 patients. The tumor regression grade (TRG), using the Mandard scoring system, was TRG1 in 16 patients (43.2%), TRG2 in 12 (32.4%), TRG3 in 12 (32.4%), TRG4 in 6 (16.2%), and TRG5 in 1 patient (2.7%). CONCLUSION: Raltitrexed plus oxaliplatin combined with pelvic radiotherapy was effective and well tolerated in patients with resectable T3 rectal carcinoma. PMID- 15337550 TI - Survivin expression is an independent prognostic factor in rectal cancer patients with and without preoperative radiotherapy. AB - PURPOSE: Survivin, as an inhibitor of apoptosis, is undetectable in normal tissues but expressed in tumors. Survivin expression in rectal cancer patients who have undergone preoperative radiotherapy (RT) alone has not been studied. We analyzed the relationships of survivin expression to RT, clinicopathologic variables, apoptosis, and p53 expression in rectal cancer patients who participated in a trial of preoperative RT. METHODS AND MATERIALS: Survivin was immunohistochemically examined in 98 rectal tumors (74 had adjacent normal mucosa). Of 98 patients, 57 underwent surgery alone and 41 underwent RT before surgery. RESULTS: Survivin positivity was related to worse survival, independent of Dukes' stage, local and distant recurrence, differentiation, gender, age, apoptosis, and p53 expression (p = 0.02). Survivin was not associated with survival in the patients without (p = 0.08) or with (p = 0.19) RT. After RT, survivin tended to be increased in adjacent normal mucosa (p = 0.057) but not in tumors (p = 0.71). CONCLUSION: Survivin was independently related to survival in rectal cancer patients who participated in a trial of preoperative RT, but not in either treatment group (surgery alone or surgery plus RT). Whether the effect of survivin on tumors is associated with RT and further related to patient survival needs to be investigated in a larger number of patients. PMID- 15337551 TI - Another way to estimate outcome of advanced nasopharyngeal carcinoma--is concurrent chemoradiotherapy adequate? AB - PURPOSE: To evaluate a simple risk grouping system and determine whether concurrent chemoradiotherapy (CCRT) is adequate for patients with advanced nasopharyngeal carcinoma (NPC). METHODS AND MATERIALS: A total of 284 patients with 1992 American Joint Committee on Cancer (AJCC) Stage III to IV (M0) NPC were analyzed retrospectively. They were treated by either radiotherapy (RT) alone or CCRT. We divided patients into high-risk and low-risk subgroups according to our experience. High-risk patients met at least one of the following criteria: (1) nodal size >6 cm, (2) supraclavicular node metastases, (3) 1992 AJCC stage T4N2, (4) multiple neck node metastases with 1 node >4 cm. The disease extent of each patient was stratified by our risk grouping system, AJCC 1992 and 1997 staging systems. Survival analyses-including nasopharynx disease free (TS), neck disease free (NS), distant metastasis disease free (MS), overall survival (OS), and progression-free (PFS) survival curves-were compared between these three different classifications. RESULTS: According to the 1992 AJCC staging system, 80.3% (228/284) of NPC patients are Stage IV, whereas only 19.7% are Stage III. Most patients are downstaged by the 1997 AJCC staging system with 28.5% (81/284) Stage IV and 71.5% (203/284) Stage III/II. Our risk criteria stratify more even patient distribution, because 119 patients (41.9%) are assigned to the high-risk group and 165 patients (58.1%) to the low-risk group. Log-rank test of Kaplan Meier survival curves, multivariate comparison of the Cox proportional hazards model, and 3 goodness-of-fit indices validated that our risk grouping system seemed to be at least as efficacious as, or slightly superior to, the 1992 and 1997 AJCC systems. The 5-year TS (95.1% vs. 76.8%, p = 0.0012), NS (100% vs. 95.7%, p = 0.0974), MS (90.5% vs. 78.1%, p = 0.0282), OS (83.2% vs. 59.7%, p = 0.0041), and PFS (87.3% vs. 61.5%, p = 0.0003) were significantly better in patients receiving CCRT than RT alone for the low-risk group. However, the corresponding survival rates between CCRT and RT for high-risk patients were 74.9% vs. 67.6% (p = 0.2545) for TS, 92.1% vs. 86.8% (p = 0.4744) for NS, 59.7% vs. 60.0% (p = 0.5537) for MS, 55.8% vs. 46.3% (p = 0.1761) for OS, and 44.5% vs. 43.1% (p = 0.3911) for PFS, respectively. CONCLUSIONS: Concurrent chemoradiotherapy is superior to RT alone for low-risk patients but inadequate for high-risk patients. Adding neoadjuvant and/or adjuvant chemotherapy would be a reasonable approach for high-risk patients. Our risk grouping criteria are a simple and useful guide that will have important implications in the design of future therapeutic trials. PMID- 15337552 TI - Three-dimensional conformal radiotherapy versus intracavitary brachytherapy for salvage treatment of locally persistent nasopharyngeal carcinoma. AB - PURPOSE: To compare the outcomes of three-dimensional conformal radiotherapy (3D CRT) and intracavitary brachytherapy (ICBT) as salvage treatment for locally persistent nasopharyngeal carcinoma. METHODS AND MATERIALS: Between March 1994 and November 2001, a total of 117 patients with locally persistent nasopharyngeal carcinoma received salvage treatment for 2-8 weeks (median, 4 weeks) after a full course of conventional external beam RT. Of the 117 patients, 54 were salvaged with 3D-CRT (3D group) and 63 with ICBT (BT group). No statistically significant differences were found in the patient characteristics between the two groups (p >0.05). In the 3D group, the planning target volume for 3D-CRT was defined as the persistent disease plus a 5-mm margin; three to seven static conformal coplanar or noncoplanar portals were delivered for each fraction. The median salvage dose was 24 Gy (range, 18-38 Gy), with fraction size of 2.0 Gy/d. In the BT group, a median salvage dose of 20 Gy (range, 15-30 Gy) was delivered with a (192)Ir source, at 5 Gy/fraction, twice weekly. The brachytherapy dose was prescribed at a distance of 1 cm from the center of the surface as defined by the sources, irrespective of the extent of persistent disease. The actuarial rates of survival were estimated using the Kaplan-Meier method. Potential differences in the actuarial outcomes between groups were evaluated using the Mantel log-rank test. Multivariate analyses were performed with the Cox regression proportional hazards model. RESULTS: The 5-year actuarial rates of overall survival, disease-specific survival, and local failure-free survival for the 3D group and BT group were 64.50% vs. 55.78% (p = 0.33), 70.03% vs. 59.56% (p = 0.11), and 88.93% vs. 76.28% (p = 0.07), respectively. Subgroup analysis showed that the 5-year actuarial local failure-free survival rate of patients with initially diagnosed T3-T4 disease for the 3D group and BT group was 84.01% vs. 60.50% (p = 0.03). The incidence of Grade 3-4 late complications was comparable between the two groups. Multivariate analyses performed in the whole group showed that T stage at initial diagnosis and the salvage technique (3D-CRT or ICBT) were the statistically significant, independent prognostic factors for local failure-free survival (p = 0.00 and p = 0.02, respectively). CONCLUSION: 3D-CRT seemed to provide better local control than ICBT as a salvage treatment for locally persistent nasopharyngeal carcinoma, especially in patients with initially diagnosed T3-T4 disease. CT/MRI evaluation of the extent of persistent disease is recommended for technique selection of salvage RT. Patients should be cautioned about the potentially increased complications. The optional time for salvage treatment remains controversial. PMID- 15337553 TI - Phase II double-blind randomized study comparing oral aloe vera versus placebo to prevent radiation-related mucositis in patients with head-and-neck neoplasms. AB - PURPOSE: In a single-institution, double-blind, prospective, randomized trial, we determined whether oral aloe vera gel can reduce radiation-induced mucositis in head-and-neck cancer patients. METHODS AND MATERIALS: We randomized 58 head-and neck cancer patients between oral aloe vera and placebo. To be included in this Phase II protocol, patients had to be treated with radiotherapy with curative intent at Stanford University between February 1999 and March 2002. We examined patients biweekly for mucositis at 15 head-and-neck subsites and administered quality-of-life questionnaires. RESULTS: Patients in the aloe and placebo groups were statistically identical in baseline characteristics. By the end of treatment, the two groups were also statistically identical in maximal grade of toxicity, duration of Grade 2 or worse mucositis, quality-of-life scores, percentage of weight loss, use of pain medications, hydration requirement, oral infections, and prolonged radiation breaks. CONCLUSION: In our randomized study, oral aloe vera was not a beneficial adjunct to head-and-neck radiotherapy. The mean quality-of-life scores were greater in the aloe vera group, but the differences were not statistically significant. Oral aloe vera did not improve tolerance to head-and-neck radiotherapy, decrease mucositis, reduce soreness, or otherwise improve patient well-being. PMID- 15337554 TI - Phenomenologic model describing flow reduction for parotid gland irradiation with intensity-modulated radiotherapy: evidence of significant recovery effect. AB - OBJECTIVE: To develop a model describing the relationship between the parotid gland radiation dose and salivary flow reduction. Salivary function was described by the "relative flow reduction" (RFR)-a continuous variable in contrast to the traditional binary response used in normal tissue complication probability estimations. METHODS AND MATERIALS: Twenty-three patients with squamous cell carcinoma of the head and neck who were treated with intensity-modulated radiotherapy (RT) were the subject of this study. Of these patients, 19 had sufficiently long follow-up to be eligible for analysis. All were treated with curative intent, most (14 of 19) in the postoperative setting. The planning objectives were to deliver a mean dose of 50, 60, or 70 Gy, respectively, to low risk microscopic, high-risk microscopic, and gross disease areas, while maintaining a mean dose of < or =20 Gy to the spared portion of one or both parotid glands. The mean dose to all parotid glands (right and left) was 30.2 Gy. All submandibular glands received >50 Gy when not surgically removed. Whole-mouth saliva collections, including both stimulated and unstimulated saliva flow, were obtained before treatment and at regular intervals after RT. These measurements were converted to the RFR by comparing the posttreatment and pretreatment flow rates. Any follow-up flow rates greater than baseline were scored as 0 relative reduction. We used Lyman's model to relate the equivalent uniform dose to RFR at various points for each patient. The equivalent uniform dose was calculated using the linear quadratic model, with an assumed alpha/beta ratio of 3 Gy for the parotid gland. Measurements were modeled 1-3 months after RT (early) and >6 months after RT (late), and using the best and worst measurements, regardless of when measured. RESULTS: Fitting the Lyman model to RFR data of unstimulated flow revealed a statistically significant dose-complication relationship. We observed a stepwise reduction in flow, with the threshold dose D(50) at 2 Gy per fraction (D(50)) increasing from 12.4 Gy (early) to 43.9 Gy (late). For the worst and best flow measurements, the corresponding D(50) (2 Gy/fr) was 13.0 Gy and 40.1 Gy, respectively. For most stimulated flow measurements, a weak relationship was found between the RFR and equivalent uniform dose. In those cases, the model did not yield a statistically significant description of the data. However, in the case of late measurements, the relationship was statistically significant and similar to that seen in the unstimulated cases, with a D(50) (2 Gy/fr) of 47.5 Gy. CONCLUSION: We observed a strong relationship between the generalized mean parotid gland dose and RFR. The threshold dose increased markedly between the early and late measurements, indicating a statistically significant recovery effect in this tissue. Compared with unstimulated flow, the RFR for stimulated flow was not described as well by the model, because the effect of the stimulant was not included in the model. PMID- 15337555 TI - Stereotactic radiotherapy for primary lung cancer and pulmonary metastases: a noninvasive treatment approach in medically inoperable patients. AB - PURPOSE: The clinical results of dose escalation using stereotactic radiotherapy to increase local tumor control in medically inoperable patients with Stage I-II non-small-cell lung cancer or pulmonary metastases were evaluated. METHODS AND MATERIALS: Twenty patients with Stage I-II non-small-cell lung cancer and 41 patients with 51 pulmonary metastases not amenable to surgery were treated with stereotactic radiotherapy at 3 x 10 Gy (n = 19), 3 x 12-12.5 Gy to the planning target volume enclosing 100%-isodose, with normalization to 150% at the isocenter; n = 26) or 1 x 26 Gy to the planning target volume enclosing 80% isodose (n = 26). The median follow-up was 11 months (range, 2-61 months) for primary lung cancer patients and 9 months (range, 2-37 months) for patients with metastases. RESULTS: The actuarial local control rate was 92% for lung cancer patients and 80% for metastasis patients > or =1 year after treatment and was significantly improved by increasing the dose from 3 x 10 Gy to 3 x 12-12.5 Gy or 1 x 26 Gy (p = 0.038). The overall survival rate after 1 and 2 years was 52% and 32%, respectively, for lung cancer patients and 85% and 33%, respectively, for metastasis patients, impaired because of systemic disease progression. After 12 months, 60% of patients with primary lung cancer and 35% of patients with pulmonary metastases were without systemic progression. No severe acute or late toxicity was observed, and only 2 patients (3%) developed symptomatic Grade 2 pneumonitis, which was successfully treated with oral steroids. CONCLUSION: Stereotactic radiotherapy for lung tumors offers a very effective treatment option locally without significant complications in medically impaired patients who are not amenable to surgery. Patient selection is important, because those with a low risk of systemic progression are more likely to benefit from this approach. PMID- 15337556 TI - Increased cure rate of glioblastoma using concurrent therapy with radiotherapy and arsenic trioxide. AB - PURPOSE: Patients with glioblastoma multiforme (GBM) do extremely poorly despite aggressive therapy with surgery, radiotherapy (RT), and chemotherapy. In an effort to increase the efficacy of therapy for GBM, we studied the efficacy of arsenic trioxide (ATO) combined with high-dose RT in GBM cells in vitro and GBM xenograft tumors in nude mice. METHODS AND MATERIALS: Human glioblastoma cell line SNB75 cells were irradiated in vitro with doses of 0-15 Gy with or without ATO. Clonogenic assays were used to generate radiation survival curves. Intracellular reactive oxygen species and apoptosis induced by ATO and RT were measured. The therapeutic efficacy of ATO alone, local tumor RT alone, and the combined therapy was tested in nude mice bearing established s.c. SNB75 tumors. A single RT dose of 20 Gy was administered locally to tumors. ATO at 10 mg/kg was injected i.p. 10 min after RT for the in vivo experiments. RESULTS: Radiation survival curves of GBM SNB75 cells demonstrated that a dose of 0.2 microM ATO increased radiation-induced cell killing by 2 logs at 10 Gy. ATO at 1 microM decreased survival from 4 x 10(-2) after 7 Gy of RT alone to 4 x 10(-5). A time course experiment demonstrated that the greatest level of cell killing occurred when ATO was administered immediately before or within 2 hours after RT. To test the therapeutic efficacy of this combined treatment regimen in vivo, nude mice with established SNB75 GBM tumors were treated with a single local tumor dose of 20 Gy of RT with or without ATO (10 mg/kg x two doses) administered weekly. Appropriate control groups were included as well. ATO alone did not inhibit tumor growth. RT at 20 Gy alone inhibited tumor growth by 45 days, with regrowth of tumors thereafter. The combination of RT and ATO resulted in complete regression of the tumors in 4 of 5 mice without tumor regrowth for up to 4 months. The fifth mouse in the combined treatment group had a 90% reduction in tumor size without progression during the 4-month follow-up period. Furthermore, ATO alone and in combination with RT did not produce any obvious signs of toxicity. CONCLUSION: These results have demonstrated that ATO increases intracellular levels of reactive oxygen species, induces apoptosis, and enhances the radiation cell killing of GBM cells. RT combined with ATO was an effective treatment for GBM tumors in this preclinical model. These preclinical results are encouraging and provide a rationale for further study of ATO combined with RT for the treatment of GBM and other histologic types of brain cancer using a variety of RT schemes. PMID- 15337557 TI - Patterns of failure in supratentorial primitive neuroectodermal tumors treated in Children's Cancer Group Study 921, a phase III combined modality study. AB - PURPOSE: To analyze the patterns of failure in patients with supratentorial primitive neuroectodermal tumors (ST-PNETs) treated with combined modality therapy in a large, randomized, multi-institutional study. METHODS AND MATERIALS: A total of 44 prospectively staged patients with ST-PNET confirmed by central pathology review were treated in the Children's Cancer Group Study 921, which compared two chemoradiotherapy regimens. The patterns of initial sites of failure were analyzed. These were compared with the failure patterns of 188 children with posterior fossa (PF) PNETs treated in the same protocol. RESULTS: The major determinant for progression-free survival was the initial metastatic stage. The 3 year progression-free survival for M0 patients was 53% +/- 8.5% compared with 14% +/- 9.4% for M+ patients. The cumulative 5-year relapse incidence was 71.4% +/- 21% for M+ patients compared with 47.5% +/- 8.6% for M0 patients. The overall failure rate for both M0 and M+ ST-PNETs was greater than that for PF-PNETs (47.5% +/- 8.6% vs. 29.3% +/- 4.7% for M0 and 71.4% +/- 21% vs. 48.4% +/- 5.5% for M+). Failure at the primary site, either as the sole site or as a component of initial failure, was also seen more frequently in ST-PNETs than in PF-PNETs. For M0 patients, the 5-year local failure rate as a component of initial failure was 42.0% +/- 8.5% for ST-PNETs compared with 17.7% +/- 3.9% for PF-PNETs. For patients with primary tumors either in the ST or PF, the 5-year spinal axis failure rate as a component of initial failure was not significantly different statistically when compared by M stage. For M+ patients, the 5-year spinal axis failure rate as a component of initial failure was 42.9% +/- 22.8% for ST-PNETs and 34.6% +/- 5.2% for PF-PNETs. CONCLUSION: Despite aggressive combined modality therapy, ST-PNETs had high rates of failure, with M+ patients faring especially poorly. Both local and spinal failure rates remained high, indicating the need to maximize both local and regional/systemic therapies. Overall, these patients fared worse than those with high-risk PF-PNETs in terms of progression-free survival and failure rates. PMID- 15337558 TI - Impact of margin for target volume in low-dose involved field radiotherapy after induction chemotherapy for intracranial germinoma. AB - PURPOSE: We previously published a report stating that germinomas with elevated serum beta human chorionic gonadotropin (HCG-beta) had a poor relapse rate, but these findings have not been supported by a multi-institutional trial. The margin for initial gross tumor volume (GTV) before surgery and chemotherapy of the same materials was investigated by retrospective review. METHODS AND MATERIAL: The 27 patients reported on in the previous paper were analyzed. The two-dimensional margin from the initial GTV to 90% of the prescribed dose of 24 Gy was 2.0 cm for a solitary lesion in the protocol. This margin was measured retrospectively without knowledge of the serum HCG-beta level. The whole ventricle field was used for patients with multifocal disease and whole central nervous system field was used for disseminated disease, respectively. RESULTS: Six relapses were seen in 18 patients with solitary tumors, and were treated with the minimum margin of 1.5 cm or less to the initial GTV. Five of the 6 had initially elevated serum HCG beta at the median of 7.4 mIU/mL, ranging from 0.7-233 mIU/mL. No relapses were seen in the 9 patients who were treated with whole ventricle or whole central nervous system field. CONCLUSIONS: An inadequate margin and elevated serum HCG beta were equally determined to be candidates that caused the poor local control. The whole ventricle is recommended as the smallest target volume for germinoma with or without elevated HCG-beta after induction chemotherapy. PMID- 15337559 TI - Radiosurgery for treatment of brain metastases: estimation of patient eligibility using three stratification systems. AB - PURPOSE: To compare three patient stratification systems predicting survival: recursive partitioning analysis (RPA), score index for radiosurgery in brain metastases (SIR), and a proposed basic score for brain metastases (BS-BM). METHODS AND MATERIALS: We analyzed the outcome of 110 patients treated with Leksell Gamma Knife radiosurgery between December 1999 and January 2003. The BS BM was calculated by evaluating three main prognostic factors: Karnofsky performance status, primary tumor control, and presence of extracranial metastases. RESULTS: The median survival was 27.6 months for RPA Class I, 10.7 months for RPA Class II, and 2.8 months for RPA Class III (p <0.0001). Using the SIR, the median survival was 27.7, 10.8, 4.6, and 2.4 months for a score of 8-10, 5-7, 4, and 0-3, respectively (p <0.0001). The median survival was undefined in patients with a BS-BM of 3 (55% at 32 months) and was 13.1 months for a BS-BM of 2, 3.3 months for a BS-BM of 1, and 1.9 months for a BS-BM of 0 (p <0.0001). The backward elimination model in multivariate Cox analysis identified SIR and BS-BM as the only two variables significantly associated with survival (p = 0.031 and p = 0.043, respectively). CONCLUSION: SIR and BS-BM were the most accurate for estimating survival. They were specific enough to identify patients with short survival (SIR 0-3 and BS-BM 0). Because of it simplicity, BS-BM is easier to use. PMID- 15337560 TI - Acoustic neuroma radiosurgery with marginal tumor doses of 12 to 13 Gy. AB - PURPOSE: To define tumor control and clinical outcomes of radiosurgery to marginal tumor doses of 12-13 Gy for unilateral acoustic neuroma patients. METHODS AND MATERIALS: Three hundred thirteen patients with previously untreated unilateral acoustic neuromas (vestibular schwannomas) underwent gamma knife radiosurgery between February 1991 and February 2001 with marginal tumor doses of 12-13 Gy (median, 13 Gy). Median follow-up was 24 months (maximum, 115 months; 36 patients with > or =60 months). Maximum doses were 20-26 Gy (median, 26 Gy), and treatment volumes were 0.04-21.4 mL (median, 1.1 mL). RESULTS: The actuarial 6 year clinical tumor control rate (no requirement for surgical intervention) for the entire series was 98.6 +/- 1.1%. Two patients required tumor resection; one had a complete resection for solid tumor growth and one required partial resection for an enlarging adjacent subarachnoid cyst. Six-year actuarial rates for preservation of facial nerve function, normal trigeminal nerve function, unchanged hearing level, and useful hearing were 100%, 95.6 +/- 1.8%, 70.3 +/- 5.8%, and 78.6 +/- 5.1%, respectively. The risk of developing trigeminal neuropathy correlated with increasing tumor volume (p = 0.038). CONCLUSIONS: Acoustic neuroma radiosurgery with doses of 12-13 Gy provides high rates of tumor control and cranial nerve preservation. PMID- 15337561 TI - Immunohistochemical study of cell cycle-associated proteins in adenocarcinoma of the uterine cervix treated with radiotherapy alone: P53 status has a strong impact on prognosis. AB - PURPOSE: Because the incidence of adenocarcinoma of the uterine cervix has recently risen, the evaluation of radiotherapy (RT) for this disease has become an increasingly urgent matter. We analyzed the expression of the cell cycle associated proteins p53, p27, p21/waf1/cip1, and cyclin D1 in cervical adenocarcinomas in correlation with the prognostic significance in tumors treated with RT alone. METHODS AND MATERIALS: The expression of p53, p27, p21/waf1/cip1, and cyclin D1 was studied using an immunohistochemical method in 53 cases of cervical adenocarcinoma treated only with RT. Patients received RT alone between 1965 and 1994. The mean patient age was 61.8 +/- 12.6 years (range, 36-82 years). The number of patients with Stage I, II, III, and IVA disease was 6, 16, 28, and 3, respectively. RESULTS: The number of patients with p53, p27, p21/waf1/cip1, and cyclin D1 positive tumors was 24, 18, 22, and 8, respectively; no statistically significant correlation was noted. The 5-year disease-free survival rate of p53-positive patients was 30%, significantly lower than the 62% for the p53-negative patients (p = 0.02); no statistically significant correlation was noted between disease-free survival and p27, p21/waf1/cip1, and cyclin D1 expression. No statistically significant correlation was observed between local control and expression of any of the proteins. CONCLUSION: Expression of p53 protein has a statistically significant impact on disease-free survival in adenocarcinoma of the uterine cervix treated with RT alone. However, the clinical significance of p27, p21/waf1/cip1, and cyclin D1 protein expression was not obvious. PMID- 15337562 TI - Prediction of outcome of advanced cervical cancer to thermoradiotherapy according to expression profiles of 35 genes selected by cDNA microarray analysis. AB - PURPOSE: To identify a set of genes related to thermoradiosensitivity of cervical carcinoma and to establish a predictive method. METHODS AND MATERIALS: A total of 19 patients with cervical cancer (1 with Stage IIIA, 11 with Stage IIIB, 5 with Stage IVA, and 2 with Stage IVB) who underwent definitive thermoradiotherapy between May 1995 and August 2001 were included in this study. We compared the expression profiles of 8 thermoradiosensitive and 11 thermoradioresistant tumors obtained by punch biopsy before treatment using a cDNA microarray consisting of 23,040 human genes. RESULTS: We selected 35 genes on the basis of a clustering analysis and confirmed the validity of these genes with a cross-validation test. Some of these genes were already known to be associated with apoptosis (BIK, TEGT, SSI-3), hypoxia-inducible genes (HIF1A, CA12), and tumor cell invasion and metastasis (CTSL, CTSB, PLAU, CD44). We developed a "predictive score" system that could clearly separate the thermoradiosensitive group from the thermoradioresistant group. CONCLUSION: These results from the treatment program between May 1995 and August 2001 showed that by using gene-expression profiles we can predict the outcome of thermoradiotherapy for advanced cervical carcinoma. A "predictive score" system was developed that could clearly separate the thermoradiosensitive group from the thermoradioresistant group. These results may eventually lead to the achievement of "personalized therapy" for this disease. PMID- 15337563 TI - Recurrent squamous cell carcinoma of cervix after definitive radiotherapy. AB - PURPOSE: To study retrospectively the characteristics and survival of patients with recurrent squamous cell carcinoma (SCC) of the cervix after definitive radiotherapy (RT) and to identify subsets of patients who might benefit from aggressive salvage treatment. METHODS AND MATERIALS: Between 1990 and 1999, 1292 patients with Stage I-IVA SCC of the cervix underwent full-course RT. Of the 1292 patients, 375 (29%) had either local or distant failure and were included in this analysis. The 35 patients (2.7%) with both pelvic and distant relapse were excluded. In the 162 patients with local failure, 71 (44%) had persistent disease and 91 (56%) had a relapse after complete tumor regression. Of these 162 patients, 47 (29%) received salvage surgery. In the 213 patients with distant failure, 46 (22%) had isolated para-aortic lymph node (PALN) metastasis, and 35 (76%) of them were treated with concurrent chemoradiotherapy/RT. Patients with supraclavicular lymph node (SCLN) relapse usually underwent concurrent chemoradiotherapy. Palliative chemotherapy and/or RT were given by decision of the responsible attending physician. RESULTS: The independent prognostic factors for local failure were advanced stage and young age (<45 years) and, for distant failure, were advanced stage, positive pelvic lymph nodes, and high serum SCC antigen levels. The 5-year overall survival rate was 10% and 11%, respectively, for patients with local or distant failure. For local relapse, the 5-year overall survival rate was 29% vs. 3% (p = 0.0001) for patients with vs. without salvage surgery and 22% vs. 9% vs. 4% for patients with tumors confined to the cervix, tumors extending but not beyond the cervix and adjacent tissues, and tumor extending beyond adjacent tissues but contained within the pelvis (p = 0.005). The survival rates, either with or without salvage surgery, were nearly identical between patients with persistent disease and those with relapse after complete regression. The 3-year overall survival rate was 34%, 28%, and 5% (p = 0.001), respectively, for patients with PALN relapse alone, SCLN relapse with or without PALN relapse, and relapse other than PALN and SCLN. Of the patients with PALN relapse alone, 27% survived >5 years. CONCLUSION: For recurrent SCC of the cervix after RT, patients with isolated PALN relapse salvaged by RT or combined chemoradiotherapy or those with cervical relapse salvaged by surgery can achieve long-term survival. Early detection of relapse with aggressive salvage treatment is important for achieving better outcome. Patients with persistent disease or relapse after complete remission had similar outcomes. Patients with SCLN relapse had a longer survival time than those with other metastases (except PALN), and palliative RT might be beneficial. PMID- 15337564 TI - Use of the small pelvic field instead of the classic whole pelvic field in postoperative radiotherapy for cervical cancer: reduction of adverse events. AB - PURPOSE: We examined whether use of small pelvic (SP) field encompassing only the pericervical regions and upper stream lymphatic will reduce the adverse events that occur with classic whole pelvic (WP) field, in postoperative radiotherapy (RT) for cervical cancer. METHODS AND MATERIAL: This retrospective study included 72 patients treated with SP field (SP group) used specifically for node-negative status and 46 patients treated with WP field (WP group) used conventionally for node-positive status. Total dose was 50.0 or 50.4 Gy at 2.0 or 1.8 Gy per fraction. Acute adverse events (nausea, diarrhea, cystitis, and leukopenia) and late adverse events (lymphedema, cystitis, ileus, and diarrhea) were graded according to the Common Toxicity Criteria and compared between groups. RESULTS: Diarrhea (Grades 2-3) and leukopenia (Grades 1-3) occurred significantly more often in WP group (32.4% and 80.5%, respectively) than in SP group (9.2% and 52.2%, respectively). Among the late events, lymphedema occurred most often overall (5-year rate: SP, 47.0%; WP, 49.1%). Only ileus occurred at a significantly higher rate in The WP group than in SP group (5-year rate: 16.2% vs. 3.2%). CONCLUSIONS: Use of the SP field tailored for node-negative status was suggested to reduce adverse events involving the intestine and hemopoietic system. PMID- 15337565 TI - Late effects of radiotherapy for pediatric extremity sarcomas. AB - PURPOSE: To determine the long-term effects of radiotherapy (RT) in children treated for extremity sarcoma. PATIENTS AND METHODS: Between 1964 and 1997, 15 of 33 children treated with RT for extremity sarcomas at the University of Iowa have survived with a median follow-up was 20 years (range, 6-36 years). There were 10 boys and 5 girls with a median age of 13 years (range, 3.5-20 years) at the time of irradiation. The diagnosis was Ewing's sarcoma in 8 (53%), synovial sarcoma in 4 (27%), alveolar rhabdomyosarcoma in 2, and fibrosarcoma in 1. Location of primary tumor was lower extremity in 10 (67%) and upper extremity in 5 (33%). RT was given as the definitive therapy for 9 children (median dose, 55.8 Gy; range, 45-66 Gy) and as an adjuvant postoperative treatment in 6 (median, 63 Gy; range, 41.4-66.4 Gy). (60)Co was used in 6 (40%), 4 mV in 4, 6 mV in 2, and 250 kV photons in 2 patients; 1 child was treated with a combination of 12 and 15 MeV electrons for a Ewing's sarcoma of the distal femur. Another child had a 25 Gy intraoperative RT boost after 41.4 Gy conventional RT. Late effects to the muscle, soft tissue, and growing bone were assessed using the objective portion of the LENT-SOMA scale proposed by the Late Effects Consensus Conference. RESULTS: Late effects were seen in all patients and included atrophy in 12 (80%), fibrosis in 12 (80%), bone growth abnormalities in 10 (67%), impairment of mobility and extremity function in 6 (40%), edema in 3 (20%), and peripheral nerve injury in 2 (13%). Ten of 15 (67%) children had Grade 1 or 2 growing bone, muscle, soft tissue, or peripheral nerve complications. Two patients (13%) had a Grade 3 mobility and extremity function score and had moderate to severe limitation of movement. Two children (13%) required epiphysiodesis because of a shorter treated leg. The patient who received an intraoperative RT boost of 25 Gy developed sensory dysfunction of the ulnar nerve 11 years after RT. Another developed radial nerve palsy 3 years after marginal resection and postoperative RT and required tendon transfer repair. One patient had radiation-induced vasculitis with popliteal artery thrombosis 23 years after RT. Five (33%) developed a fracture of the irradiated bone at a median time of 8 years after RT (range: 9 months to 22.2 years); all had Ewing's sarcoma, and 3 of these patients were subsequently found to have a secondary bone cancer (osteosarcoma 2, malignant fibrous histiocytoma 1) in the RT field. One of these patients also developed breast cancer 26 years after lung RT for metastatic Ewing's sarcoma. Overall, 11 surgical procedures in 8 children were performed to correct a limb preservation treatment toxicity. CONCLUSIONS: Although most children treated with RT for a pediatric extremity sarcoma have minimal late toxicity by LENT-SOMA scale, approximately half required a surgical procedure to correct a late effect. A fracture in the irradiated bone may be the presenting sign or may precede a radiation-induced bone malignancy, as seen in 3 of the patients in this study. PMID- 15337566 TI - Plasma citrulline concentration: a surrogate end point for radiation-induced mucosal atrophy of the small bowel. A feasibility study in 23 patients. AB - PURPOSE: Plasma citrulline, a nitrogen end product of glutamine metabolism in small-bowel enterocytes, was suggested as a marker of radiation-induced small bowel epithelial cell loss in mice after single-dose whole-body irradiation. Our objective was to evaluate the feasibility of citrulline as a marker for radiation induced small-intestinal mucosal atrophy in patients during and after abdominal fractionated radiotherapy. METHODS AND MATERIALS: Twenty-three patients were studied weekly during treatment and at intervals of 2 weeks and 3 and 6 months after treatment by postabsorptive plasma citrulline concentration and clinical toxicity grading. The interrelationship between these variables and the correlation with small-bowel dose and volume parameters were investigated. RESULTS: During fractionated radiotherapy, citrulline concentration significantly decreased as a function of the radiation dose (p < 0.001) and the volume of small bowel treated (p = 0.001). The plasma citrulline concentration correlated with clinical toxicity during the last 3 weeks of treatment. As a whole, citrulline concentration correlated better with radiation dose and volume parameters than clinical toxicity grading. CONCLUSIONS: In patients treated with fractionated radiation therapy for abdominal or pelvic cancer sites, plasma citrulline concentration may be a simple objective marker for monitoring epithelial cell loss, a major event in acute radiation-induced small-bowel toxicity. PMID- 15337567 TI - Evaluation of clinical utility of BTC-2000 for measuring soft tissue fibrosis. AB - PURPOSE: To evaluate whether mechanical tissue parameters, specifically laxity (in millimeters) and energy absorption (millimeters of mercury multiplied by millimeters) as measured by the BTC-2000, could discriminate levels of fibrosis severity among patients treated for extremity soft tissue sarcoma by surgery alone; preoperative radiotherapy (RT) and surgery; and surgery followed by postoperative RT. METHODS AND MATERIALS: A total of 41 patients were treated for extremity soft tissue sarcoma by surgery alone (n = 11); preoperative RT (50 Gy in 2-Gy daily fractions) and surgery (n = 15); and surgery followed by postoperative RT (66 Gy in 2-Gy daily fractions; n = 15). Serial fibrosis measurements were evaluated at equal intervals from the midpoint of the surgical incision along the length of the incision. On the basis of the average of these measurements, differences among the three groups were analyzed using analysis of variance. RESULTS: Pair-wise statistically significant differences were found among the three treatment groups for both laxity and energy absorption as determined by the average of all measurements. The treatment difference remained statistically significant even after adjusting for differences based on the untreated contralateral limb and anatomic site (p <0.001 and p = 0.002 for laxity and energy absorption, respectively). CONCLUSION: The biomechanical tissue parameters of laxity and energy absorption discriminated fibrosis severity in patients treated with different RT doses. The BTC-2000 may provide a useful quantitative measure of soft tissue fibrosis. PMID- 15337568 TI - Radiation-induced otitis media--study of a new test, vestibular-evoked myogenic potential. AB - PURPOSE: Excluding the radiation history, current physical examination and audiovestibular function tests fail to differentiate radiation-induced otitis media (ROM) from chronic otitis media (COM). This study applied the newly developed vestibular-evoked myogenic potential (VEMP) test to investigate whether the VEMP test can be of help in differentiating between them. METHODS AND MATERIALS: Fourteen irradiated nasopharyngeal carcinoma (NPC) patients with ROM (18 ears) and 14 age-matched, and gender-matched patients with COM (18 ears) were enrolled. Each patient underwent stimulation with a short tone burst initially, and then by tapping the forehead using a tendon hammer. To define the effect of the neck soft tissue on the VEMP response further, an additional two groups (10 non-NPC patients with ROM but no neck fibrosis/edema and 10 non-NPC patients without ROM but with neck fibrosis/edema) were included for comparison. RESULTS: The occurrence of normal VEMPs in the ROM ears (33%) did not differ significantly from that in the COM ears (56%) by tone-burst stimulation. However, using tapping evocation, most (89%) COM ears revealed normal VEMPs, and most (61%) ROM ears demonstrated delayed VEMPs. Furthermore, most (90%) non-NPC patients with ROM but no neck fibrosis/edema revealed delayed or absent VEMPs. In contrast, all non-NPC patients with neck fibrosis/edema but no ROM demonstrated normal VEMPs bilaterally, implying that the effect of neck soft tissue on the VEMP response is less. CONCLUSION: Through tapping evocation, most COM ears revealed normal VEMPs, and most ROM ears demonstrated delayed VEMPs, indicating that ROM is different from COM because of the larger affected areas such as retrolabyrinthine or brainstem involvement. This result can explain why grommet insertion or tympanoplasty is not beneficial to postirradiated ears, possibly because both operations spare the inflammation outside the middle ear cavity. PMID- 15337569 TI - Assessment and management of cutaneous reactions with amifostine administration: findings of the ethyol (amifostine) cutaneous treatment advisory panel (ECTAP). AB - PURPOSE: To review reports of severe skin reactions during amifostine treatment. METHODS AND MATERIAL: The expert panel reviewed postmarketing reports of skin reactions and discussed strategies for evaluation and management. RESULTS: Between 1994 and April 2002, 35 events were classified as severe skin reactions worldwide: erythema multiforme (8), Stevens-Johnson syndrome (10), toxic epidermal necrolysis (11), toxicoderma (3), and bullae (3). Unadjusted incidences were 6-9 per 10,000 radiotherapy patients and 0.8-1 per 10,000 chemotherapy patients. In 10 patients (29%) amifostine was continued after cutaneous signs and symptoms appeared. CONCLUSIONS: Practical recommendations for practicing clinicians were developed. Cutaneous evaluation for rash, ulceration, or lesions particularly on lips/mucosa, palmar/plantar surfaces, and the trunk-should be performed before amifostine administration. Reactions can be classified as local injection site/radiation port reactions or non-injection site reactions; and non injection site reactions with associated fever or constitutional symptoms must be differentiated from radiation-induced dermatitis or cutaneous reaction with another etiology. Amifostine should be permanently discontinued for severe skin reactions or reactions associated with constitutional symptoms not known to be due to any other etiology. Increased physician awareness, proper patient management, monitoring before administration, and early intervention/discontinuation for non-injection site reactions may reduce the incidence of cutaneous reactions and enhance amifostine safety. PMID- 15337570 TI - Effects of nicotinamide and carbogen in different murine colon carcinomas: immunohistochemical analysis of vascular architecture and microenvironmental parameters. AB - PURPOSE: To investigate oxygenation, perfusion, and cell proliferation in two murine colon carcinoma lines with known differences in chemotherapy sensitivity and analyze the effect of nicotinamide and carbogen on these tumor characteristics. METHODS AND MATERIALS: Mice with s.c. transplanted C38 and C26a murine colon tumors were treated with nicotinamide and carbogen and compared with control tumors. Two markers of hypoxia, CCI-103F and pimonidazole, were injected before and after treatment with nicotinamide/carbogen, respectively, allowing each tumor to serve as its own control. Hoechst33342 was used as a perfusion marker and bromodeoxyuridine (BrdUrd) as a proliferation marker. Frozen tumors were cut for multistep immunostaining and computer-controlled microscope scanning for hypoxic fractions (HF), perfused fractions (PF), vascular density, and BrdUrd labeling index (LI). RESULTS: Microscopic observation of C38 and C26a tumors showed extensive differences in vascular architecture, distribution patterns of hypoxia, and BrdUrd-labeling. Quantitative analysis of C38 and C26a tumors showed a decrease in HF in response to all treatment modalities. For C38 tumors, the average decrease in HF in response to carbogen containing treatments was larger than to nicotinamide alone. In C26a tumors, no difference in average decrease in HF was observed between the treatments. The PF of C38 and C26a did not change in response to treatment. The LI of C38 and C26a decreased upon all treatments, which was statistically significant in the combination treatment of C38. CONCLUSIONS: The mechanism that can simultaneously explain all the observed changes in response to treatment may be the conversion of metabolism from less respiration toward more glycolysis due to increased glucose levels (Crabtree effect), although other mechanisms of actions cannot be excluded. PMID- 15337571 TI - The importance of postoperative CT image fusion verification of stereotactic interstitial irradiation for brain tumors. AB - PURPOSE: The aim of our study was verification of the position of implanted catheters with (125)I seeds after the catheter implantation for the brachytherapy of brain tumors. METHODS AND MATERIALS: The fusion of the CT image used at planning and after the implantation of the catheters enabled us to verify the position of the catheters containing the isotopes. After this, the tumor volume covered by the prescribed dose (TV(PD)) and the normal tissue volume covered by the prescribed dose (NTV(PD)) were compared between the plan and the actual result. The image fusion was performed by the BrainLab-Target 1.19 software on an Alfa 430 (Digital) workstation. RESULTS: The position of the catheters was adjusted in 14 (20%) of the 70 image fusion cases being studied. The position of 16 of the 116 catheters (13.8%) required adjustment after the fusion of control images in the 70 cases studied. The Student t probe revealed a significant difference between the TV(PD) values of the reality and the plans (75.8% vs. 92.4%, p < 0.0001). There was a significant difference between values of the real performances and planning for NTV(PD) (86.8% vs. 76%, p = 0.001) and for the conformity index (0.37 vs. 0.54, p = 0.0001), too. CONCLUSION: The application of the interstitial irradiation with CT control allows us to identify and correct possible inaccuracies in catheter positioning during the operation. The procedure then becomes far more accurate and reliable, and as such, the irradiation becomes more conformal than without catheter adjustment. PMID- 15337572 TI - On-line aSi portal imaging of implanted fiducial markers for the reduction of interfraction error during conformal radiotherapy of prostate carcinoma. AB - PURPOSE: An on-line system to ensure accuracy of daily setup and therapy of the prostate has been implemented with no equipment modification required. We report results and accuracy of patient setup using this system. METHODS AND MATERIALS: Radiopaque fiducial markers were implanted into the prostate before radiation therapy. Lateral digitally reconstructed radiographs (DRRs) were obtained from planning CT data. Before each treatment fraction, a lateral amorphous silicon (aSi) portal image was acquired and the position of the fiducial markers was compared to the DRRs using chamfer matching. Couch translation only was used to account for marker position displacements, followed by a second lateral portal image to verify isocenter position. Residual displacement data for the aSi and previous portal film systems were compared. RESULTS: This analysis includes a total of 239 portal images during treatment in 17 patients. Initial prostate center of mass (COM) displacements in the superior, inferior, anterior, and posterior directions were a maximum of 7 mm, 9 mm, 10 mm and 11 mm respectively. After identification and correction, prostate COM displacements were <3 mm in all directions. The therapists found it simple to match markers 88% of the time using this system. Treatment delivery times were in the order of 9 min for patients requiring isocenter adjustment and 6 min for those who did not. CONCLUSIONS: This system is technically possible to implement and use as part of an on-line correction protocol and does not require a longer than standard daily appointment time at our center with the current action limit of 3 mm. The system is commercially available and is more efficient and user-friendly than portal film analysis. It provides the opportunity to identify and accommodate interfraction organ motion and may also permit the use of smaller margins during conformal prostate radiotherapy. Further integration of the system such as remote table control would improve efficiency. PMID- 15337573 TI - Feasibility of synchronization of real-time tumor-tracking radiotherapy and intensity-modulated radiotherapy from viewpoint of excessive dose from fluoroscopy. AB - PURPOSE: Synchronization of the techniques in real-time tumor-tracking radiotherapy (RTRT) and intensity-modulated RT (IMRT) is expected to be useful for the treatment of tumors in motion. Our goal was to estimate the feasibility of the synchronization from the viewpoint of excessive dose resulting from the use of fluoroscopy. METHODS AND MATERIALS: Using an ionization chamber for diagnostic X-rays, we measured the air kerma rate, surface dose with backscatter, and dose distribution in depth in a solid phantom from a fluoroscopic RTRT system. A nominal 50-120 kilovoltage peak (kVp) of X-ray energy and a nominal 1-4 ms of pulse width were used in the measurements. RESULTS: The mean +/- SD air kerma rate from one fluoroscope was 238.8 +/- 0.54 mGy/h for a nominal pulse width of 2.0 ms and nominal 100 kVp of X-ray energy at the isocenter of the linear accelerator. The air kerma rate increased steeply with the increase in the X-ray beam energy. The surface dose was 28-980 mGy/h. The absorbed dose at a 5.0 cm depth in the phantom was 37-58% of the peak dose. The estimated skin surface dose from one fluoroscope in RTRT was 29-1182 mGy/h and was strongly dependent on the kilovoltage peak and pulse width of the fluoroscope and slightly dependent on the distance between the skin and isocenter. CONCLUSION: The skin surface dose and absorbed depth dose resulting from fluoroscopy during RTRT can be significant if RTRT is synchronized with IMRT using a multileaf collimator. Precise estimation of the absorbed dose from fluoroscopy during RT and approaches to reduce the amount of exposure are mandatory. PMID- 15337574 TI - What can we conclude from the results of an out-of-date breast brachytherapy study? In regard to Poti Z, Nemeskeri C, Fekeshazy A, et al. (Int J Radiat Oncol Biol Phys 2004;58:1022-1033). PMID- 15337576 TI - In regard to Poti et al.: Partial breast irradiation with interstitial (60)co brachytherapy results in frequent grade 3 or 4 toxicity: Evidence based on a 12 year follow-up of 70 patients (Int J Radiat Oncol Biol Phys 2004;58:1022-1033). PMID- 15337578 TI - Effects of organotin compounds on pubertal male rats. AB - Tributyltin (TBT) and triphenyltin (TPT) induce effects in male and female reproductive organs of rodents. They also cause tumors in these organs and it is theorized that they result from endocrine disruption. We studied the effects of 40 mg methyltestosterone (MTT), 0.5 or 15 mg TBT and 2, 6 or 12 mg TPT/kg bw on the male sexual development using a modification of the Rodent 20-Day Thyroid/Pubertal Male Assay. Male Wistar rats were treated per gavage for 30 days beginning at 23 days of age. A delay in the completion of preputial separation was observed after administration of MTT and 15 mg/kg TBT. Changes in weights of one or more reproductive organs were observed in all treatment groups. Testosterone concentration was decreased in the MTT, the 15 mg TBT as well as in the 6 and 12 mg TPT groups. A decrease in luteinizing hormone (LH) concentration was observed in the MTT and 15 mg TBT groups while an increase was seen after exposure to 6 mg TPT/kg bw. We conclude that peripubertal exposure to 15 mg TBT and 6 mg TPT/kg bw clearly affected male sexual development. PMID- 15337579 TI - Effects of a novel organophosphorus pesticide (RPR-V) on extra hepatic detoxifying enzymes after repeated oral doses in rats. AB - The effects of a novel organophosphorous pesticide, 2-butenoic acid-3-(diethoxy phosphinothionyl) ethyl ester (RPR-V) on glutathione S-transferases (GST), UDP glucuronyl transferases (UDPGT) and the level of glutathione (GSH) were evaluated in rats after repeated oral administration at 33 microg kg(-1)day(-1) (low), 66 microg kg(-1)day(-1) (mid) and 99 microg kg(-1)day(-1) (high) for 90 days and at 28 days (withdrawal) post-treatment. GSH level and GST in kidney; GSH level in brain decreased significantly at mid and high doses on 45th and 90th day (P < 0.05). However, UDPGT activity in brain and kidney increased significantly at mid and high doses on 45th and 90th day (P < 0.05). Interestingly, the withdrawal study revealed that the effect was reversible 28 days after the treatment when all the enzymes reverted to levels close to those of controls. These results suggest that RPR-V has the potential to modulate the extrahepatic detoxifying enzymes and thereby interact with other physiological processes in the exposed organisms. PMID- 15337580 TI - Intermediates of Krebs cycle correct the depression of the whole body oxygen consumption and lethal cooling in barbiturate poisoning in rat. AB - Rats poisoned with one LD50 of thiopental or amytal are shown to increase oxygen consumption when intraperitoneally given sucinate, malate, citrate, alpha ketoglutarate, dimethylsuccinate or glutamate (the Krebs cycle intermediates or their precursors) but not when given glucose, pyruvate, acetate, benzoate or nicotinate (energy substrates of other metabolic stages etc). Survival was increased with succinate or malate from control groups, which ranged from 30-83% to 87-100%. These effects were unrelated to respiratory depression or hypoxia as judged by little or no effect of succinate on ventilation indices and by the lack of effect of oxygen administration. Body cooling of comatose rats at ambient temperature approximately 19 degrees C became slower with succinate, the rate of cooling correlated well with oxygen consumption decrease. Succinate had no potency to modify oxygen consumption and body temperature in intact rats. A condition for antidote effect of the Krebs intermediate was sufficiently high dosage (5 mmol/kg), further dose increase made no odds. Repeated dosing of succinate had more marked protective effect, than a single one, to oxygen consumption and tended to promote the attenuation of lethal effect of barbiturates. These data suggest that suppression of whole body oxygen consumption with barbiturate overdose could be an important contributor to both body cooling and mortality. Intermediates of Krebs cycle, not only succinate, may have a pronounced therapeutic effect under the proper treatment regimen. Availability of Krebs cycle intermediates may be a limiting factor for the whole body oxygen consumption in barbiturate coma, its role in brain needs further elucidation. PMID- 15337581 TI - Fumonisin B1-induced toxicity and oxidative damage in U-118MG glioblastoma cells. AB - The mycotoxin fumonisin B1 (FB1) is produced by Fusarium verticillioides, which commonly infects corn and other agricultural products. Fusarium species are also a frequent finding in moisture-damaged buildings, causing possible human exposure to FB1. FB1 is neurotoxic and carcinogenic in a number of animal species. In this study, we have investigated the effects of FB1 on human U-118MG glioblastoma cells. The production of reactive oxygen species (ROS), lipid peroxidation, intracellular reduced glutathione (GSH) levels, cell viability, caspase-3-like protease activity and DNA fragmentation were studied in cells exposed to 0.01-100 microM FB1 for 0.5-144 h. FB1 increased lipid peroxidation and the production of ROS in U-118MG cells, showing significant effects after culture times from 48 to 144 h at dose levels of 10 or 100 microM FB1. These effects were accompanied by changes in the GSH levels and cell viability, which decreased significantly after incubating the cells for 48-144 h with the toxin. Signs of apoptosis were indicated by increased caspase-3-like protease activity and internucleosomal DNA fragmentation. Thus, oxidative stress and apoptosis may be involved in the neurotoxicity induced by FB1. PMID- 15337582 TI - In utero exposure to low-dose 2,3',4,4',5-pentachlorobiphenyl (PCB 118) impairs male fertility and alters neurobehavior in rat offspring. AB - Neurobehavior (motor activity and developmental reflexes) and male reproductive parameters were evaluated in rat offspring after in utero exposure to a low dose of PCB 118 comparable to human exposure levels. Sprague-Dawley dams were treated on gestation day 6 by gavage with a single dose of 375 microg PCB 118/kg body weight or peanut oil (control). The dose was calculated to be approximately 100 fold higher than that found in human breast milk. Postnatal reflexes, motor activity and male reproductive performance were evaluated in rat offspring after exposure to PCB 118. Evaluation of locomotor activity for five consecutive days during puberty (PND 70-74) revealed hyperactivity in offspring from PCB 118 exposed dams. In adult males (PND 170), clear effects on reproductive organs were observed in PCB-exposed animals which had smaller testes, epididymides and seminal vesicles (absolute and relative weights). Decreases in sperm and spermatid numbers and impairment of daily sperm production were also observed. Our results clearly demonstrate that low-dose exposure to PCB 118 alters neurobehavior and impairs adult male fertility in offspring. This is in contrast to the reported increases in sperm production and testis weight in rat after high dose PCB exposures. PCBs appear to possess variable dose-related effects and therefore low-dose studies are important to obtain a complete picture for human risk assessment. PMID- 15337583 TI - Cytotoxicity of occupationally and environmentally relevant mycotoxins. AB - Mycotoxins can cause various toxic effects in humans. Acute and chronic respiratory diseases were reported after inhalation of organic dust containing toxigenic moulds and mycotoxins, respectively. To gain first insights into health effects from airborne exposure to these compounds, five toxigenic airborne moulds of the genera Aspergillus and Penicillium collected at composting plants and eight reference mycotoxins were tested for cytotoxicity in four established cell lines as a surrogate of tissues known or suspected to be targets of toxic effects of mycotoxins. The known mycotoxins sterigmatocystin, fumagillin, verruculogen, penitrem A, and roquefortine C were detected in extracts of the moulds. All five extracts caused serious toxic effects in the cell lines. Sterigmatocystin caused a 80-fold higher toxicity in the A-549 lung cell line compared to Hep-G2 liver cells indicating a specific susceptibility of A-549 to this agent. Since only a minor part of the toxic effects of the extracts in A-549 cells and--to a lesser extent--in the other cell lines could be explained by contents of the identified mycotoxins, the presence of additional mycotoxins or other toxic principles is assumed in the mould extracts. However, the detected mycotoxins in the mould extracts and their distinctive cytotoxicity support the hypothesis that mycotoxins may be involved in the aetiology of lung diseases due to the inhalation of organic dust. PMID- 15337584 TI - Epigallocatechin gallate prevents oxidative-stress-induced death of mutant Cu/Zn superoxide dismutase (G93A) motoneuron cells by alteration of cell survival and death signals. AB - This study was undertaken to evaluate the effect of the G93A mutation in the human Cu/Zn-superoxide dismutase gene (hSOD1) on the phosphatidylinositol-3 kinase (PI3K)/Akt and glycogen synthase kinase-3 (GSK-3) pathway in motoneuron, and to determine the role of epigallocatechin gallate (EGCG) on oxidative stress injured motoneurons. The viability of G93A mutant cells was less than that of wild-type cells, and the activation of PI3K and the phosphorylation of Akt and GSK-3 in G93A mutant cells decreased compared with wild-type hSOD1 4.1 cells. In the experiment to evaluate the effect of oxidative stress and/or EGCG on these motoneurons, after exposure to 400 microM H2O2, the MTT assay revealed greatly reduced viability of G93A mutant cells compared with wild-type cells, and pre treatment of these cells with EGCG before H2O2 exposure increased the viability of both cell lines. Western blot analysis showed that the G93A mutation and oxidative stress decreased survival signals including PI3K/Akt but increased death signals including GSK-3; however, pre-treatment with EGCG increased survival signals but decreased death signals. These results suggest that PI3K/Akt and GSK-3 activities are altered in G93A mutant cells and EGCG-induced activation of PI3K/Akt and inhibition of GSK-3 could be a new potential therapeutic strategy for ALS associated with oxidative injury. PMID- 15337585 TI - Endosulfan-induced cardiotoxicity and free radical metabolism in rats: the protective effect of vitamin E. AB - Endosulfan is widely used in insect control and it is absorbed by both humans and animals through ingestion, inhalation and percutaneously. The aim of this work was to study antioxidant enzyme system which include superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) and malondialdehyde (MDA), the end product of lipid peroxidation and ultrastructural changes that might occur in the heart tissue of adult male Wistar rats as a result of endosulfan intoxication. Vitamin E (200 mg/kg, twice a week), endosulfan (2 mg/kg, per day, once a day in corn oil) and vitamin E (200 mg/kg, twice a week)+endosulfan (2 mg/kg, per day, once a day in corn oil) combination were given to rats (n = 10/group) orally via gavage for 6 weeks. SOD, GPx, CAT activities and MDA level increased in the endosulfan-treated group heart tissue compared to control group (P < 0.01, P < 0.01, P < 0.05 and P < 0.01, respectively). SOD, GPx activities and MDA level decreased in the vitamin E + endosulfan-treated group compared to endosulfan-treated group (P < 0.05, P < 0.05 and P < 0.05, respectively). Decrease of CAT activity was not significant statistically in the vitamin E + endosulfan-treated group compared to endosulfan-treated group. CAT activity increased in the vitamin E + endosulfan treated group compared to control group (P < 0.05). Increase of SOD, GPx activities and MDA levels were not significant statistically in the vitamin E + endosulfan-treated group compared to control group. In electron microscopic investigations while cytoplasmic edema and swelling and vacuolization of mitochondria of myocardial cells in endosulfan treated group was observing, only a weak swelling of mitochondria of myocardial cells in vitamin E + endosulfan-treated group was observed. We conclude that vitamin E significantly reduce endosulfan-induced cardiotoxicity in rats. PMID- 15337586 TI - Degradation of nonylphenol polyethoxylates by ultraviolet B irradiation and effects of their products on mammalian cultured cells. AB - Nonylphenol polyethoxylates (NPEOs) are widely used as non-ionic surfactants and their biodegradation products such as 4-n-nonylphenol are stable and have been demonstrated to be cytotoxic. In the aquatic environment, these compounds are usually exposed to sunlight, and while the correlation between the biodegradation of NPEOs and changes in cytotoxicity has been reported, the relationship between the photodegradation of NPEOs and cytotoxicity has not. In this study, we investigated the degradation of NPEO by ultraviolet (UV) irradiation, especially UVB irradiation, and the effects on mammalian cell lines. NPEO with a smaller number of ethylene oxide (EO) units showed greater cytotoxicity. Although NPEO (10) completely inhibited the proliferation of the cells, NPEO (70) showed no toxicity. UVB irradiation significantly induced a shortening of the side chain, which was due to the production of ROS. The EO side chain of NPEO (10), was gradually degradated, but that of NPEO (70) was degradated near the benzene ring. Furthermore, the degradation of the benzene ring was more effective in NPEO (70) than NPEO (10). The toxicity of NPEO (10) in cultured cells decreased following UVB irradiation, whereas that of NPEO (70) was induced after UVB irradiation at 500 J/cm2 and disappeared at 1000 J/cm2. This might be due to the production of NPEO with a short side chain and 4-n-nonylphenol by the degradation of EO and due to the degradation of the benzene ring at higher doses of UVB irradiation. This study shows the significance of UV exposure to the degradation of alkylphenol polyethoxylates in the environment. PMID- 15337587 TI - Modulation of aryl hydrocarbon receptor-regulated gene expression by arsenite, cadmium, and chromium. AB - Aryl hydrocarbon receptor (AhR) ligands and heavy metals are environmental co contaminants and their molecular interaction may disrupt the coordinated regulation of AhR-dependent phase I and II drug metabolizing enzymes. To determine the effect of heavy metals on the AhR-regulated genes: cytochrome P4501A1 (Cyp1a1), NAD(P)H: quinone oxidoreductase (QOR) and glutathione S transferase Ya (GST Ya), murine hepatoma Hepa 1c1c7 cells were treated with increasing concentrations of As3+ (1-10 microM), Cd2+ (1-25 microM) and Cr6+ (1 25 microM) with or without the AhR ligands: 2,3,7,8-tetrachlorodibenzo-p-dioxin (0.1 nM), 3-methylcholanthrene (0.25 microM), beta-naphthoflavone (10 uM), or benzo[a]pyrene (1 microM). Our results show that AhR ligands alone and As3+ or Cd2+ alone increased the catalytic activities and mRNA levels of all AhR regulated genes. When metals were co-administered with an AhR ligand, all three metals inhibited the induction of Cyp1a1 activity by the AhR ligands but potentiated its mRNA and protein expression. In addition, all metals enhanced QOR and GST Ya at the activity and mRNA levels but modulated their induction by AhR ligands in a concentration, metal, and AhR ligand-dependent manner. Generally, Cr6+ inhibited while As3+ and Cd2+ potentiated the induction of QOR and GST Ya activities and mRNA levels. The three metals enhanced the expression of heme oxygenase-1, which coincided with the changes in the phase I and phase II enzyme activities. These results show that the ability of metals to alter the capacity of AhR ligands to induce the bioactivating phase I and the detoxifying phase II enzymes will influence the carcinogenicity and mutagenicity of the AhR ligands. PMID- 15337589 TI - The European Sporotrichiella project: a polyphasic approach to the biology of a new Fusarium species. PMID- 15337590 TI - Fusarium langsethiae sp. nov. on cereals in Europe. AB - A new species of Fusarium, Fusarium langsethiae, is described, illustrated and discussed. This species is isolated from kernels of oats, wheat and barley in several European countries. Morphologically, the species resembles Fusarium poae. It is differentiated from F. poae by slower growth, less aerial mycelium and absence of odour; its napiform or globose conidia are borne in the aerial mycelium on the agar surface on often bent phialides which exhibit sometimes more than one opening, whereas those of F. poae are produced on straight monophialides mostly in the aerial mycelium. No sporodochial conidia are formed by F. langsethiae even under near-UV light (nUV). Based on morphological characters, the species is placed in the section Sporotrichiella. PMID- 15337591 TI - Diversity in metabolite production by Fusarium langsethiae, Fusarium poae, and Fusarium sporotrichioides. AB - The production of mycotoxins and other metabolites by 109 strains of Fusarium langsethiae, Fusarium poae, Fusarium sporotrichioides, and F. kyushuense was investigated independently in four laboratories by liquid or gas chromatography analyses of cultural extracts with UV diode array, electron capture, or mass spectrometric detection systems. From the compiled results, it was found that F. langsethiae consistently produced the trichothecenes diacetoxyscirpenol (DAS), T 2 toxin (T-2), HT-2 toxin (HT-2), and neosolaniol (NEO) and, to a lesser extent, some additional trichothecene derivatives. F. langsethiae also produced culmorins, chrysogine (CHRYS), aurofusarin (AUF), and enniatin (EN). F. sporotrichioides showed a metabolite profile similar to that of F. langsethiae, while F. poae had a different profile as 41 of 49 strains produced nivalenol (NIV) and other 8-keto trichothecenes, in addition to DAS and derivatives of this metabolite. Only a trace amount of NIV was detected from one strain of F. kyushuense. In summary, all the three core taxa of this joint study were found to produce trichothecenes. Fusarin C (F-C) was not detected from F. langsethiae, but it was produced by F. poae and F. sporotrichioides. Aurofusarin was only detected from a few strains of F. langsethiae, while nearly all strains of F. poae and F. sporotrichioides produced this compound. In contrast, chrysogine was not detected from F. poae, but was produced by the other two taxa. Production of enniatins was scattered among the three main taxa of this study, whereas beauvericin (BEA) was produced by many strains of F. poae and F. sporotrichioides. Only one odd strain of F. langsethiae (IBT 9959) produced beauvericin. However, the status of this strain is uncertain. By a polyphasic approach using species-specific metabolite profiles, the fruity odour of F. poae, and morphological observations, it was concluded that F. langsethiae, F. poae, and F. sporotrichioides should be regarded as three significant taxa at a species level. PMID- 15337592 TI - Phylogenetic relationship of Fusarium langsethiae to Fusarium poae and Fusarium sporotrichioides as inferred by IGS, ITS, beta-tubulin sequences and UP-PCR hybridization analysis. AB - Fusarium langsethiae was recently described to accommodate "powdery" isolates of Fusarium poae, which morphologically resemble F. poae, but whose metabolite profile is similar to that of Fusarium sporotrichioides. In order to investigate the phylogenetic relationship of F. langsethiae to closely related species, we sequenced the internal transcribed spacer (ITS) regions 1 and 2 and part of the intergenic spacer (IGS) region of the rDNA cluster and part of the beta-tubulin gene from 109 strains of F. poae, F. sporotrichioides, F. langsethiae and Fusarium kyushuense from different geographic origin. Sequence analysis of ITS1 and 2 was unable to separate all F. sporotrichioides strains from F. langsethiae strains. Sequence analysis of beta-tubulin distinguished all four species, but it did not resolve the phylogenetic relationship between these two species. Sequence analysis of the IGS region distinguished the four species and led to a higher number of subgroups of the individual species, of which that of F. sporotrichioides var. minus isolates was even better supported than that of F. poae and F. langsethiae. Neighbor-joining and POY analyses of all combined sequences reliably separated all species studied, including F. langsethiae, clearly from F. sporotrichioides. The high intraspecific variability of the IGS sequences were found useful to group isolates according to their geographic origin. These results are in accordance with the results of the UP-PCR hybridization analysis. In summary, our data offer molecular support for the description of F. langsethiae as a new species in section Sporotrichiella. PMID- 15337593 TI - Phylogenetic analyses of the Fusarium poae, Fusarium sporotrichioides and Fusarium langsethiae species complex based on partial sequences of the translation elongation factor-1 alpha gene. AB - Phylogenetic relationships between four Fusarium species were studied using parts of the nuclear translation elongation factor-1 alpha (EF-1alpha) gene as a phylogenetic marker. Sequences from 12 isolates of Fusarium poae, 10 isolates of Fusarium sporotrichioides and 12 isolates of Fusarium langsethiae yielded 4, 5 and 5 haplotypes, respectively. In addition, we included one isolate of Fusarium kyushuense. The aligned sequences were subjected to neighbor-joining (NJ), maximum parsimony and maximum likelihood (ML) analyses. The results from the different analyses were highly concordant. The EF-1alpha-based phylogenies support the classification of F. langsethiae as a separate taxon in the section Sporotrichiella of Fusarium, as the closest sister taxon to F. sporotrichioides, while F. kyushuense is the sister taxon to F. poae. This corresponds well with the ability of F. langsethiae and F. sporotrichioides to produce T-2 and HT-2 toxins. In contrast, morphological characters indicate a closer relationship between F. langsethiae and F. poae on the one hand, and between F. sporotrichioides and F. kyushuense on the other hand. PMID- 15337594 TI - AFLP analysis of Fusarium species in the section Sporotrichiella-evidence for Fusarium langsethiae as a new species. AB - Amplified fragment length polymorphism (AFLP) was applied to compare 100 isolates of Fusarium poae, Fusarium sporotrichioides, and Fusarium langsethiae. Comparison of fingerprints revealed several markers specific for each group of isolates. Cluster analysis showed a clear separation of F. poae from F. sporotrichioides and F. langsethiae. F. langsethiae isolates formed a rather homogeneous group separated from F. sporotrichioides. The AFLP data obtained support F. langsethiae as a new Fusarium species, for the time being in the section Sporotrichiella. PMID- 15337595 TI - The use of tri5 gene sequences for PCR detection and taxonomy of trichothecene producing species in the Fusarium section Sporotrichiella. AB - Purified DNA from isolates of Fusarium poae, Fusarium sporotrichioides, Fusarium kyushuense and Fusarium langsethiae was used as a template to amplify a 658-bp fragment from the trichodiene synthase (tri5) gene of these fungi with the gene specific PCR primer pair Tox5-1/Tox5-2. Fragments obtained were isolated and sequenced. DNA sequence alignments revealed high similarity between the sequences derived from F. sporotrichioides and F. langsethiae (98.7%) and less similarity between the latter species and F. poae (90.9%). Phylogenetic analysis of the aligned sequences using the tri5 sequence of Fusarium pseudograminearum as an outgroup revealed clear separation between one group consisting of F. poae and F. kyushuense and another consisting of F. sporotrichioides and F. langsethiae. The two latter species could not be distinguished phylogenetically on the basis of their tri5 sequences. Taxon-specific reverse primers were designed from the aligned sequences and combined with the tri5 gene-specific forward primer Tox5-1. The new reverse primers enabled specific amplification of a fragment of approximately 400 bp from DNA isolated from F. sporotrichioides, F. poae, F. langsethiae and F. kyushuense, respectively. All primers were tested for cross reactivity with DNA from 26 fungal species potentially capable of producing trichothecenes. Only the primer designed for F. langsethiae cross-reacted with F. sporotrichioides. PCR assays were applied in analysis of artificially and naturally infected samples of barley and oats. On artificially infected barley, species were selectively detected by the corresponding primers. In naturally infected oats, F. langsethiae was identified by the combination of two PCR assays designed for detection of F. sporotrichioides and F. langsethiae, respectively. PMID- 15337596 TI - IGS-RFLP analysis and development of molecular markers for identification of Fusarium poae, Fusarium langsethiae, Fusarium sporotrichioides and Fusarium kyushuense. AB - The intergenic spacer (IGS) regions of the rDNA of several Fusarium spp. strains obtained from the collaborative researchers (Int. J. Food Microbiol. (2003)) were amplified by polymerase chain reaction (PCR), and an IGS-RFLP analysis was performed. Restriction digestion with AluI, MspI and PstI allowed differentiation between the related Fusarium poae and Fusarium kyushuense species. Fusarium langsethiae was also separated from Fusarium sporotrichioides (including var. minus) on the basis of the banding patterns after MspI digestion, while specific XhoI, AluI and MspI restriction patterns were found in the IGS amplicons of F. sporotrichioides var. minus. According to the phylogenetic analysis of IGS-RFLP patterns, F. langsethiae (except for one strain), F. sporotrichioides, F. poae and F. kyushuense strains formed four well-supported clades with high-bootstrap values. Based on the sequence differences in the IGS region, species-specific primers were designed for the F. langsethiae/F. sporotrichioides group and for F. poae. The specificity and sensitivity of the primers were tested on various Fusarium species and isolates, and on several other important fungal genera associated with cereals. The F. poae-specific primers, designed in this study, showed the same specificity as primers Fp82f/Fp82r developed previously. The two phylogenetic subgroups of F. langsethiae, found by IGS sequencing analysis, were separated on the basis of size differences of the amplification products with primers CNL12/PulvIGSr specific for the F. langsethiae/F. sporotrichioides group. RFLP analysis of the amplified IGS region is a useful molecular assay for characterisation and a phylogenetic study of several related Fusarium species-F. langsethiae, F. sporotrichioides, F. sporotrichioides var. minus, F. poae and F. kyushuense. The primers designed in this study were highly specific and allowed identification of F. poae and the F. langsethiae/F. sporotrichioides group. PMID- 15337597 TI - Specific detection of Fusarium langsethiae and related species by DGGE and ARMS PCR of a beta-tubulin (tub1) gene fragment. AB - Fusarium langsethiae was recently described as a new toxigenic Fusarium species, which morphologically resembles Fusarium poae, but exhibits a mycotoxin pattern related to Fusarium sporotrichioides. To develop tools for early and specific detection of F. langsethiae and distinguishing it from related species of section Sporotrichiella and Discolor (F. poae, F. sporotrichioides, Fusarium kyushuense, Fusarium robustum, Fusarium sambucinum and Fusarium tumidum) sequence variations in their beta-tubulin-encoding (tub1) gene were employed to design two PCR-based methods, denaturing gradient gel electrophoresis (DGGE) and amplification refractory mutation system (ARMS)-PCR. DGGE reliably separated all these strains, even from mixtures and in the presence of DNA from their natural hosts Zea mais, Triticum aestivum and Avena sativa. In addition, a tetraprimer ARMS-PCR, which employs two primer pairs to amplify, respectively, two different fragments of tub1 in a single PCR reaction resulted in rapid differentiation between F. langsethiae, F. sporotrichioides and F. poae according to the number of amplicons (four, two and one, respectively). These two methods will thus be worthwhile tools in the specific detection of F. langsethiae in infected crops. PMID- 15337598 TI - An integrated taxonomic study of Fusarium langsethiae, Fusarium poae and Fusarium sporotrichioides based on the use of composite datasets. AB - An integrated systematic study was carried out to clarify the taxonomical position and relationship of Fusarium langsethiae to other taxa within the Fusarium section Sporotrichiella. Strains of this species were compared with strains of the closely related species Fusarium poae and Fusarium sporotrichioides using a composite dataset. This set consisted of DNA sequences derived from the ribosomal internal transcribed spacer (ITS) regions, partial sequences of the ribosomal intergenic spacer (IGS) region, the beta-tubulin and translation elongation factor-1 alpha (EF-1alpha) genes, AFLP fingerprints, chromatographic data on secondary metabolites and morphological data and growth characteristics. From these combined data, a consensus matrix was calculated by taking the mean of all pairwise distances between single isolates over all separate datasets. The consensus matrix was used as the basis for the construction of a UPGMA dendrogram and a multidimensional scaling, both of which revealed a clear separation of the three taxa. Partial IGS, EF-1alpha and beta tubulin sequence-as well as chromatography-and AFLP-derived similarities turned out to be comparably consistent, while ITS sequence- and morphology-derived similarity matrices were rather divergent. PMID- 15337599 TI - The coordination of imidazole and substituted pyridines by the hemeoctapeptide N acetyl-ferromicroperoxidase-8 (FeIINAcMP8). AB - The N-terminus acetylated ferric hemeoctapeptide from cytochrome c, N acetylmicroperoxidase-8 (Fe(III)-NAcMP8) can be reduced by dithionite in aqueous solution to produce Fe(II)-NAcMP8. The UV-Vis spectrum has a broad Soret band and relatively poorly defined Q bands which is consistent with a mixture of a five coordinate high spin species with His as the axial ligand and a six-coordinate, predominantly high spin species with His/H(2)O as axial ligands. There are two spectroscopically observable pK(a)s at 8.7+/-0.1 and 10.9+/-0.2 which are attributed to ionization of a heme propionic acid group and coordinated H(2)O, respectively; a pK(a) > or = 14 is due to ionization of the proximal His ligand. Equilibrium constants were determined by UV-Vis spectrophotometry at 25.0+/-0.2 degrees C and 0.5 M ionic strength (NaClO(4)) for the coordination of imidazole and a number of substituted pyridines, and complement available data for the ferric hemepeptide, allowing a comparison to be made of the affinity of an iron porphyrin with Fe in the +2 and +3 oxidation states towards these ligands. Imidazole is coordinated more strongly by the ferric porphyrin (log K=4.08) than by the ferrous porphyrin (log K=3.40). The equilibrium constants for coordination of pyridines by the ferric and ferrous porphyrins increase and decrease, respectively, with increasing ligand basicity. Values determined by cyclic voltammetry show the same dependence on the identity of the ligand. In the ferric porphyrin, the stability of the complex increases with the basicity of the ligand and hence its ability to donate electron density onto the metal. In the case of the more electron rich ferrous porphyrin, greater stability occurs with pyridine ligands that have an electron withdrawing group and hence can accept electron density from the metal. This is consistent with the midpoint reduction potentials E(1/2) of the pyridine complexes determined by cyclic voltammetry; E(1/2) is linearly dependent on, and becomes more negative with an increase in, ligand basicity. Log K for coordination of pyridines by the ferrous hemepeptide correlates well with the energy of the ligand frontier orbital with pi symmetry, suggesting that pi-bonding effects are significant in determining the strength of binding of pyridines by a ferrous porphyrin. PMID- 15337600 TI - Copper(I) interaction with model peptides of WD6 and TM6 domains of Wilson ATPase: regulatory and mechanistic implications. AB - With the aim to investigate the mechanism of Cu(I) transport by Wilson ATPase (ATP7B), we have studied the interaction of the peptides 2K10p (CH(3)CO-Lys-Gly Met-Thr-Cys-Ala-Ser-Cys-Val-His-Asn-Lys-CONH(2)), and 2K8p (CH(3)CO-Lys-Leu-Cys Ile-Ala-Cys-Pro-Cys-Ser-Lys-CONH(2)), part of the sixth metal binding domain (WD6) and the sixth transmembrane segment (TM6) of Wilson ATPase, respectively, by means of CD, NMR spectroscopy and homology modeling. In addition, the interaction of Cu(I) with the 2K8p mutants 1s (CH(3)CO-Lys-Leu-Ser-Ile-Ala-Cys Pro-Cys-Ser-Lys-CONH(2)), 2s (CH(3)CO-Lys-Leu-Cys-Ile-Ala-Ser-Pro-Cys-Ser-Lys CONH(2)) and 3s (CH(3)CO-Lys-Leu-Cys-Ile-Ala-Cys-Pro-Ser-Ser-Lys-CONH(2)), containing two cysteines in various positions, have been studied with the same methods, in order to understand the role of each cysteine in copper binding. Our studies show that the three cysteine thiolates present in the 2K8p peptide sequence act mainly as bridging ligands for Cu(I) binding, and dithiothreitol acts as an important ligand in Cu(I) ligation by 2K10p and the 2K8p mutants. Formation of oligomeric species has been evidenced for all peptides except 2s. Shift of the equilibrium between the various oligomeric species has been accomplished by reducing the Cu(I):peptide ratio. Significant shifts of proline protons upon interaction with Cu(I) have been observed for all proline containing peptides implying a possible role of proline in facilitating Cu(I) binding. These findings have been further discussed with respect to the molecular basis of copper trafficking and intermolecular interactions. PMID- 15337601 TI - Interaction of copper with cysteine: stability of cuprous complexes and catalytic role of cupric ions in anaerobic thiol oxidation. AB - Copper complexes with cysteine have been investigated by optical spectroscopy, NMR and ESR. Cuprous ions strongly bind to the thiol group of Cys forming polymeric species with bridging thiolate sulfur according to a stoichiometry of about 1:1.2 and stability constant of the order of 10(10) M(-1). Cupric ions in the presence of cysteine, up to a ratio 0.45:1, are reduced to Cu(I) with stoichiometric production of cystine. The Cu(I) produced by this reaction is complexed by the excess of Cys. Trace amounts of Cu(II) exceeding the ratio 0.45:1 induce fast and complete oxidation of the Cys-Cu(I) complex to cystine with concomitant production of Cu(0) which precipitates. The experimental data are consistent with a mechanism by which Cu(II) oxidizes the complex Cys-Cu(I) to cystine producing aqueous Cu(I) which undergoes dismutation regenerating Cu(II). According to this mechanism the uncomplexed Cu(II) plays a catalytic oxidative role in the absence of molecular oxygen. The biological significance of these reactions is discussed. PMID- 15337602 TI - Resonance Raman study of deoxy and ligated (O2 and CO) mesoheme IX-reconstituted myoglobin, hemoglobin and its alpha and beta subunits. AB - In this work, we corrected the resonance Raman (RR) results presented earlier for deoxy mesoheme IX-reconstituted hemoglobin (mesoHb) alpha and beta subunits implied that mesohemes in these subunits undergo substantial structural changes upon formation of a hemoglobin tetramer (Biochemistry 29 (1990) 5087). We show that these data were probably due to the improper handling of the deoxy mesoheme subunit preparation. Additionally, we discuss the RR spectra of deoxy, oxy, and CO species of mesoheme IX-reconstituted myoglobin (mesoMb) and alpha and beta deoxy meso hemoglobin subunits, including their analogues with deuterium substituted mesoheme IX in all methyl groups (d(12)). Based on the obtained data, we propose a complete RR band assignment for all of the investigated molecules. The most pronounced changes are observed for the gamma(7) mode (out-of-plane movement of methane carbon atoms) associated with the interaction of the ethyl groups with the globin. We also show that in mesoheme IX-reconstituted proteins, the O(2) molecule binds stronger than in the case of native species. This is manifested by the up-shift of nu(Fe-O(2)). PMID- 15337603 TI - Identification of putative transcription factor binding sites in rodent selenoprotein W promoter. AB - To understand transcriptional regulation of the selenoprotein W (SeW) gene, we used in vitro binding assays to identify transcription factors that may be involved in the transcriptional regulation of the SeW gene. Using protein from rat C6 (glial) cell nuclear extracts, oligonucleotides containing putative regulatory elements in the SeW promoter and antibodies, we observed that specificity protein 1(Sp1) transcription factor binds to the Sp1 consensus sequence in the SeW promoter as well as to the metal response element (MRE). Although competition analysis showed specific binding at the TFII-1 site, super shift analysis using anti-TFII-1 antibody did not yield any super-shifted band. Therefore, the SeW gene may be a target for Sp1 whose binding to various regulatory sequences of the SeW promoter may activate or repress the transcription of SeW. The MRE, GRE, AP-1 and LF-A1 sites were also tested but no evidence was obtained for specific binding as indicated by lack of competition with unlabeled probes. PMID- 15337604 TI - Comparison of the predicted in vivo behaviour of the Sn(II)-APDDMP complex and the results as studied in a rodent model. AB - In a quest for more effective radiopharmaceuticals for pain palliation of metastatic bone cancer, this paper relates results obtained with ((117m)Sn labelled) Sn(II) complexed to the bone seeking bisphosphonate, N,N dimethylenephosphonate-1-hydroxy-3-aminopropylidenediphosphonate (APDDMP). APDDMP is synthesised from the known bone cancer pain palliation agent 1-hydroxy-3 aminopropylidenediphosphonate (APD, Pamindronate). This work is performed to utilise the idea that the low bone marrow radio toxicity of (117m)Sn could afford a highly effective radiopharmaceutical in pain palliation but also in the curative treatment of bone metastasis. Complex-formation constants of APDDMP with the important blood plasma metal-ions, Ca(2+), Mg(2+), Zn(2+) as well as the added metal ion, Sn(2+) were measured by glass electrode potentiometry at 25 degrees C and I = 150 mM. Blood plasma models were constructed using the computer code ECCLES and the results compared with those gathered from tests on a rodent model. The ((117m)Sn-labelled) Sn(II)-APDDMP complex was found to have only some liver and bone uptake although a high trabecular to normal bone ratio was recorded. From the blood plasma model this was shown to be primarily due to the high affinity of APDDMP for Ca(II) causing some of the Sn(II)-APDDMP complex to dissociate. High kidney uptake and excretion as well as high bladder uptake was recorded which was shown to be due to the dissociation of the Sn(II)-APDDMP complex in blood plasma. Animal model observations could be explained by the blood plasma modelling. PMID- 15337605 TI - Intercalation of Zn(II) and Cu(II) complexes of the cyclic polyamine Neotrien into DNA: equilibria and kinetics. AB - The equilibria and kinetics of the interaction of the Zn(II) and Cu(II) complexes of the macrocyclic polyamine 2,5,8,11-tetraaza[12]-[12](2,9)[1,10] phenanthrolinophane (Neotrien) with calf thymus DNA have been investigated at pH=7.0 and T=25 degrees C by spectrophotometry, spectrofluorimetry and stopped flow method. At low dye/polymer ratios both complexes bind to DNA according to the excluded site model. At high dye/polymer ratios the binding displays cooperative features. The logarithm of the binding constant depends linearly on log[NaCl]. The kinetic results suggest the D + S <==> D, S <==> DS mechanism where the metal complexes (D) react with the DNA sites (S) leading to fast formation of an externally bound form (D,S) which, in turn, is converted into internally bound complex (DS) by intercalation. The binding constants, evaluated as ratios of rate constants, agree with those obtained from equilibrium binding experiments, thus confirming the validity of the proposed model. Fluorescence titrations, where the metal-Neotrien complexes were added to DNA previously saturated with ethidium bromide (EB), show that both complexes displace EB from the DNA cavities. The reverse process, i.e. the addition of excess ethidium to the DNA/metal Neotrien systems, leads to fluorescence recovery for DNA/ZnNeotrien but not for DNA/CuNeotrien. This observation suggests that the binding of CuNeotrien induces deep alterations in the DNA structure. Experiments with Poly(dA-dT)*Poly(dA-dT) and Poly(dG-dC)*Poly(dG-dC) reveal that CuNeotrien mainly affects the structure of the latter polynucleotide. PMID- 15337606 TI - Protonation status of metal-bound ligands can be determined by quantum refinement. AB - The protonation status of key residues and bound ligands are often important for the function of a protein. Unfortunately, protons are not discerned in normal protein crystal structures, so their positions have to be determined by more indirect methods. We show that the recently developed quantum refinement method can be used to determine the position of protons in crystal structures. By replacing the molecular-mechanics potential, normally used in crystallographic refinement, by more accurate quantum chemical calculations, we get information about the ideal structure of a certain protonation state. By comparing the refined structures of different protonation states, the one that fits the crystallographic raw data best can be decided using four criteria: the R factors, electron density maps, strain energy, and divergence from the unrestrained quantum chemical structure. We test this method on alcohol dehydrogenase, for which the pK(a) of the zinc-bound solvent molecule is experimentally known. We show that we can predict the correct protonation state for both a deprotonated alcohol and a neutral water molecule. PMID- 15337607 TI - Cobaltocene-mediated catalytic monooxygenation using holo and heme domain cytochrome P450 BM3. AB - The feasibility of replacing NADPH with 1,1'-dicarboxycobaltocene in the catalytic cycle of cytochrome P450 BM3 has been explored. Using the holoprotein, the surrogate mediator was observed to reduce both the FAD and FMN in the reductase domain, as well as the iron in the heme domain. In an electrochemical system, the mediator was able to support lauric acid hydroxylation at a rate of 16.5 nmol product/nmol enzyme/minute. Similar electron transfer and catalysis were observed for the heme domain alone in the presence of the metallocene; the turnover rate in this case was 1.8 nmol product/nmol enzyme/minute. Parallel studies under the same conditions using a previously reported cobalt sepulchrate mediator showed that the two systems give similar results for both the holoenzyme and the heme domain. PMID- 15337608 TI - Neurologic disease, falls and fractures. PMID- 15337609 TI - Carotid intima-media thickness and cognitive decline: what does it mean for prevention of dementia? PMID- 15337610 TI - Risk factors for hip fracture among elderly patients with Alzheimer's disease. AB - Incidence of hip fracture among patients with Alzheimer's disease (AD), especially in elderly patients, is high. To analyze risk factors of hip fracture, we prospectively studied a cohort of elderly female patients with AD. Subjects studied were 225 female patients with AD, and the average age was 76 years old. At baseline, we recorded body mass index (BMI), a score of Mini-Mental State Examination (MMSE) and bone mineral density (BMD), and measured serum concentrations of ionized calcium, intact parathyroid hormone (PTH), pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP), intact bone Gla protein (BGP), 25-hydroxyvitamin (25-OHD) and 1, 25-dihydroxyvitamin D (1, 25 [OH]2D). The patients were followed for 2 years. During the 2-year study, hip fractures occurred in 29 patients. We compared baseline variables between the 29 patients with and 176 patients without hip fracture. AD patients with lower BMD, low concentrations of serum ionized calcium and 25-OHD (mean 3.0 ng/ml) with compensatory hyperparathyroidism were found to have an increased risk of hip fracture. Also, concentrations of serum ICTP and BGP were higher in the fracture group than in the nonfracture group. Elderly female AD patients with low BMD and serum 25-OHD concentrations <5 ng/ml with secondary hyperparathyroidism have a high risk of hip fracture, and the risk may be reduced by vitamin D supplementation. PMID- 15337611 TI - Expression of the interferon-alpha/beta-inducible MxA protein in brain lesions of subacute sclerosing panencephalitis. AB - The type-I interferon (IFN) inducible human MxA protein exhibits antiviral activity against a variety of RNA viruses including the measles virus (MV). In this study, we investigated the association between the expression of MV antigens and MxA in subacute sclerosing panencephalitis (SSPE) brains. We analyzed the MxA expression in and around lesions in brains of three SSPE patients and compared it with normal brains. Double staining with antibodies against MxA and the MV nucleocapsid revealed that MxA was highly expressed in a belt surrounding MV antigen-positive lesions in SSPE brains. In normal appearing regions distant from a lesion in SSPE brains and in normal brains, MxA was not detected. Furthermore, MxA was often less or not expressed in the center of lesions expressing high amounts of MV antigens. Such a pattern of MxA expression in SSPE brains clearly indicates that newly infected cells release type I IFN and will become demarcated by a protecting barrier of MxA expressing cells. Double staining with antibodies against MxA and glial fibrillary acidic protein (GFAP) showed that the MxA protein was expressed mainly in the cytoplasm of astrocytes. MxA expression did not correlate with the presence of cellular infiltrates of inflammatory cells, although some lymphoid cells were also positive for MxA. Since MxA inhibits the replication of MV, these findings suggest that the IFN-induced MxA protein plays an important role in slowing down the viral spread in SSPE brains and by doing so may contribute to the persistence of the MV-infection. PMID- 15337612 TI - Electromyogram-triggered neuromuscular stimulation and stroke motor recovery of arm/hand functions: a meta-analysis. AB - Debate persists about the effectiveness of poststroke behavioral interventions for progress toward motor recovery. The current meta-analysis assessed the effect of electromyogram (EMG)-triggered neuromuscular stimulation on arm and hand functions. Computer searches of PubMed and Cochran databases, as well as hand searches of reference lists identified seven EMG-triggered neuromuscular stimulation studies. Outcome measures focused on arm and hand motor capability functions. In addition, the quality of each study was rated on three guidelines: randomization, double blind, and dropouts. After adjusting data for consistency in the arm/hand outcome measures and to avoid bias, five active stimulation studies were included in the analysis. Rehabilitation treatment in each study focused on wrist extension. The total number of individuals in the treatment groups was 47 whereas the control groups had 39 subjects. The meta-analysis revealed a significant overall mean effect size (delta=0.82, S.D.=0.59). A homogeneity test indicated that the pooled standardized effect sizes estimated the same treatment effect. A fail-safe test for null effect findings revealed that 15 studies were required to reduce the large effect (0.82) to a small effect (0.20). These improved wrist extension motor capabilities findings support EMG triggered neuromuscular stimulation as an effective poststroke protocol. PMID- 15337613 TI - Common carotid artery intima media thickness and post-stroke cognitive impairment. AB - BACKGROUND AND PURPOSE: Acute stroke and other forms of cerebrovascular disease are well-recognized causes of cognitive impairment. Common carotid artery intima media thickness (CCA-IMT) has been associated with certain forms of cerebrovascular disease, but its association with cognitive impairment of vascular origin has not been elucidated. The purpose of this study was to investigate whether CCA-IMT is associated with cognitive impairment 1 year after an acute ischemic stroke. METHODS: A total of 171 consecutive patients with the first ever stroke (mean age 66+/-11.5, 41% female) underwent carotid ultrasonography during hospitalization. Demographic data, vascular risk factors and presenting stroke features were also recorded. One year later, patients' cognitive performance and depression were assessed using the Mini-Mental State Examination (MMSE), and the Montgomery Asberg Depression Rating Scale (MADRS). RESULTS: Cognitive impairment (MMSE score<24) was found in 67 (39%) of the 171 patients. CCA-IMT was significantly associated with cognitive impairment, and this association remained unchanged (OR 1.94; 95% CI 1.19-3.18) after adjustment for demographic data, vascular risk factors, stroke features, other carotid ultrasonography measurements and depression. Older age, low education level, large hemispheric lesions, hyperdense carotid plaques and depression were also independently associated with post-stroke cognitive impairment. CONCLUSIONS: In this study, CCA-IMT was independently associated with cognitive impairment 1 year after an acute ischemic stroke, and thus, it might help with the screening of stroke patients at risk of cognitive impairment. PMID- 15337614 TI - Multiparametric ISODATA analysis of embolic stroke and rt-PA intervention in rat. AB - To increase the sensitivity of MRI parameters to detect tissue damage of ischemic stroke, an unsupervised analysis method, Iterative Self-Organizing Data Analysis Technique Algorithm (ISODATA), was applied to analyze the temporal evolution of ischemic damage in a focal embolic cerebral ischemia model in rat with and without recombinant tissue plasminogen activator (rt-PA) treatment. Male Wistar rats subjected to embolic stroke were investigated using a 7-T MRI system. Rats were randomized into control (n=9) and treated (n=9) groups. The treated rats received rt-PA via a femoral vein at 4 h after onset of embolic ischemia. ISODATA analysis employed parametric maps or weighted images (T1, T2, and diffusion). ISODATA results with parametric maps are superior to ISODATA with weighted images, and both of them were highly correlated with the infarction size measured from the corresponding histological section. At 24 h after embolic stroke, the average map ISODATA lesion sizes were 37.7+/-7.0 and 39.2+/-5.6 mm2 for the treated and the control group, respectively. Average histological infarction areas were 37.9+/-7.4 mm2 for treated rats and 39.4+/-6.1 mm2 for controls. The R2 values of the linear correlation between map ISODATA and histological data were 0.98 and 0.96 for treated and control rats, respectively. Both histological and map ISODATA data suggest that there is no significant difference in infarction area between non-treated and rt-PA-treated rats when treatment was administered 4 h after the onset of embolic stroke. The ISODATA lesion size analysis was also sensitive to changes of lesion size during acute and subacute stages of stroke. Our data demonstrate that the multiparameter map ISODATA approach provides a more sensitive quantitation of the ischemic lesion at all time points than image ISODATA and single MRI parametric analysis using T1, T2 or ADCw. PMID- 15337615 TI - Decrease in GTP-sensitive high affinity agonist binding of muscarinic acetylcholine receptors in autopsied brains of dementia with Lewy bodies and Alzheimer's disease. AB - To determine changes in signal transduction from the muscarinic acetylcholine receptor (mAChR) to G protein in brain tissue of dementia with Lewy bodies (DLB) and Alzheimer's disease (AD), we investigated GTP-sensitive agonist high affinity binding, which is considered an index of the formation of the mAChR-G protein complex. Brain tissue was obtained at necropsy from eight patients with DLB, nine patients with Alzheimer's disease and seven patients as controls. Membrane fractions were prepared from frontal and temporal cerebral tissues. Displacement curves of [(3)H]l-quinuclidinyl benzilate (QNB) binding by carbamylcholine were analyzed by the nonlinear least-squares methods. The proportion of and affinity for the agonist in GTP-sensitive agonist high affinity binding were estimated. The percentages GTP-sensitive agonist high affinity bindings were significantly decreased in DLB (P<0.01) and Alzheimer's disease (P<0.05) only in the frontal lobe. There were no significant differences in the temporal lobe. The ratio of agonist affinity (Kd value of low affinity component/Kd value of high affinity component) did not significantly differ among groups in either the frontal lobe or temporal lobe. The concentration of mAChR-G protein complex is considered reduced in the frontal lobe of brains with DLB and Alzheimer's disease. Therefore, signal transduction from mAChR to G protein was disturbed in the frontal lobe in these diseases. PMID- 15337616 TI - Segregation pattern and biochemical effect of the G3460A mtDNA mutation in 27 members of LHON family. AB - Inheritance and expression of mitochondrial DNA (mtDNA) mutations are crucial for the pathogenesis of Leber hereditary optic neuropathy (LHON). We have investigated the segregation and functional consequences of G3460A mtDNA mutation in 27 members of a three-generation family with LHON syndrome. Specific activity of respiratory chain complex I in platelets was reduced in average to 56%, but no direct correlation between the mutation load and its biochemical expression was found. Heteroplasmy in blood, platelets and hair follicles varied from 7% to 100%. Segregation pattern exhibited tissue specificity and influence of different nuclear backgrounds in four branches of the pedigree. Longitudinal analysis revealed a significant (p=0.02) decrease in blood mutation load. Although enzyme assay showed reduction of complex I activity, our results give additional support to the hypothesis that expression of LHON mutation depends on complex nuclear mitochondrial interaction. PMID- 15337617 TI - Clinical characterization of idiopathic intracranial hypertension at the Detroit Medical Center. AB - BACKGROUND: Idiopathic intracranial hypertension (IIH), also known as pseudotumor cerebri (PTC), is a condition characterized by increased intracranial pressure without clinical, laboratory or radiologic evidence of an intracranial space occupying lesion, meningeal inflammation or venous outflow obstruction. Previous population-based studies evaluated the presenting clinical features of IIH in North-Eastern Libya, Iowa, Louisiana, Minnesota, Northern Ireland, Israel and Western Turkey. However, we report the demographic patterns of a metropolitan North American city. METHODS: We performed a retrospective study of all patients with an initial IIH diagnosis and follow-up at the Detroit Medical Center (DMC), Michigan from 1985 to 2002. Medical records of 174 patients were reviewed, with 77 patients fitting the diagnostic criteria. RESULTS: Seventy-seven patients, 71 females (92.2%) and 6 males (7.8%) were identified. Age ranged from 10 to 50 years, with a mean of 34 years. Fifty patients (65%) were African-American and 24 patients (31%) were Caucasian. The most common presentation was isolated headache (28.6%). Nineteen patients (24.7%) were asymptomatic. Obesity was described in 65 of 74 patients (87.8%) evaluated for body dimensions. CONCLUSIONS: We report an increased number of asymptomatic patients after routine ophthalmic examination. Obese females in metropolitan Detroit would benefit from routine eye evaluations to avoid potential visual loss related to papilledema. PMID- 15337618 TI - Redox status of plasma coenzyme Q10 indicates elevated systemic oxidative stress in Parkinson's disease. AB - Oxidative stress is suggested to play an important role in the pathogenesis of Parkinson's disease (PD). However, no elevation of plasma oxidative stress marker has been reported. We measured percent content of the oxidized form of coenzyme Q10 in total coenzyme Q10 (%CoQ-10) because %CoQ-10 has been shown to be a sensitive marker of oxidative stress. A slight but significant elevation in %CoQ 10 was observed in PD patients when compared with age/gender-matched normal subjects, suggesting elevated systemic oxidative stress in PD patients. PMID- 15337619 TI - Leptomeningeal metastasis: survival and prognostic factors in 155 patients. AB - In this single-center retrospective study, 155 consecutive patients with leptomeningeal metastasis (LM) were analyzed for the prognostic role of patient- and therapy-related variables. Ten percent of the patients received radiotherapy alone, 32% had chemotherapy alone, 31% received radiochemotherapy, 17% had supportive therapy only, and 10% were not evaluable for therapy. Chemotherapy was systemic (17%), combined systemic and intrathecal (10%), or intrathecal only (35%). Clinical improvement was noted in 41% of the patients. Overall median survival time (MST) was 4.8 months. Survival varied considerably depending on the type of primary tumor in this largest published cohort of LM patients. Univariate Cox regression analysis revealed that age >60 and elevated cerebrospinal fluid (CSF) albumin or lactate levels were therapy-independent predictors of poor survival in the entire cohort as well as in the subgroup of patients with systemic primary tumors (n=105). The assessment of three therapy-independent parameters allows to group LM patients into groups of low, intermediate, and high risk of poor survival. Moreover, the application of systemic chemotherapy was a positive prognostic factor in patients with subarachnoid lesions detected by neuroimaging (RR 1.94, p=0.001) or with extra-CNS tumor deposits (RR 1.52, p=0.05). The results of this study suggest that systemic chemotherapy alone or in combination with other therapeutic modalities may improve outcome in patients with subarachnoid tumor cell deposits detectable by neuroimaging. PMID- 15337620 TI - Dipyridamole and headache--a pilot study of initial dose titration. AB - Headache is reported by about one third of patients using dipyridamole (DP) after stroke or TIA. No study has systematically examined if initial dipyridamole titration may affect this headache. We therefore randomised patients to (1) standard aspirin and dipyridamole treatment b.i.d. for 2 weeks or (2) titration with aspirin only in the morning and aspirin and dipyridamole in the evening for 5 days, followed by 9 days of standard aspirin and dipyridamole treatment. Among 57 patients included for analysis, moderate to severe headache was reported by 28% in the standard treatment group and 25% in the titration group (n.s.). Headache for more than two consecutive days occurred in 24% and 11%, respectively. Rescue medication because of headache for more than 2 days was used by 14% and 0% in the respective groups. The cumulative number of days with headache was more than twice as high in the standard treatment group. The total numbers of other side effects were 25 and 11 in the two groups. The observed differences in this pilot study were not statistically significant, but nevertheless suggest that titration with an initially lower dose of dipyridamole may be considered to reduce headache and thereby increase compliance. A larger study is needed to clarify this with sufficient statistical power. PMID- 15337621 TI - Short-term brain atrophy changes in relapsing-remitting multiple sclerosis. AB - The objective of this study was to establish whether the time interval of 3 months is sufficient to detect whole-brain atrophy changes in patients with relapsing-remitting (RR) multiple sclerosis (MS). Another aim was to assess the value of monthly gadolinium (Gd)-enhanced magnetic resonance imaging (MRI) and of different Gd-enhancement patterns as predictors of brain atrophy. Thirty patients with RRMS (mean disease duration 4.9 years, mean age 34.4 years and mean Expanded Disability Status Scale [EDSS] 1.4) were assessed at baseline and monthly for a period of 3 months with clinical and MRI examinations. Calculations of baseline and monthly absolute and percent changes of MRI measures have been obtained using two semiautomated (Buffalo and Trieste) and one automated (SPM99) segmentation method. Changes of brain parenchymal fraction (BPF) were investigated according to Gd-enhancement patterns. Mean absolute and percent changes of BPF did not significantly differ at any time point in the study for any of the three methods. There was slight but not significant decrease of BPF from baseline to month 3: 0.0004 (0.05%), p=0.093 for Trieste; -0.0006 (0.07%), p=0.078 for Buffalo; and 0.0006 (0.08%), p=0.081 for SPM99 method. In ring-enhancement positive patients, there was a significant difference between baseline and month 3 changes of BPF, EDSS, and number of relapses. Over the study period, we did not demonstrate differences between changes of BPF according to the presence of Gd enhancement. Longitudinally, multiple regression analysis demonstrated that the only clinical or MRI parameter that predicted BPF decrease was the mean absolute change of ring enhancing lesion load (R=0.62, p=0.003). The noteworthy findings of this study are (1) the observation that a significant brain atrophy progression cannot be detected over a 3-month period in RRMS; (2) the demonstration that the ring enhancement pattern may contribute to more severe brain tissue loss in the short term; and (3) the lack of relationship between the presence and duration of Gd enhancement activity and brain volume changes in the short term. PMID- 15337622 TI - Spontaneous regression of cerebral AVM due to thrombosis of draining vein- angiographic and MRI demonstration. AB - Spontaneous regression of cerebral arteriovenous malformation (AVM) is rare and poorly understood phenomenon. We present a case of spontaneous complete and asymptomatic occlusion of the AVM associated with thrombosis of the draining cortical vein. The thrombosed draining cortical vein is beautifully demonstrated on magnetic resonance imaging (MRI). PMID- 15337623 TI - Posterior antebrachial cutaneous nerve conduction study in radial neuropathy. AB - Radial neuropathy most commonly occurs as a result of external compression at the spiral groove region. The posterior antebrachial cutaneous nerve (PACN) conduction study was performed in 15 consecutive patients with radial palsy. Unilateral PACN abnormalities were present in 11 patients. A normal PACN study was correlated with clinical improvement at 3 months. Conversely, PACN abnormality was correlated with radial motor axon loss and a poorer prognosis. The PACN study is a simple adjunct which provides additional information relating to the diagnosis and prognosis of radial lesions. PMID- 15337624 TI - Five young patients with cryptogenic stroke who used an ephedra-containing compound in a time window prior to suffering a stroke. PMID- 15337626 TI - Platelets, a typical source of error in real-time PCR quantification of mitochondrial DNA content in human peripheral blood cells. AB - Nucleoside analogues can induce toxic effects on mitochondria by inhibiting the human DNA polymerase-gamma. The clinically observed toxicities can range from slightly increased serum lactate levels to potentially severe and fatal lactic acidosis. A growing interest exists for detection of changes in mitochondrial (mt) DNA content in patients receiving antiretroviral therapy (HAART). Most studies use peripheral blood mononuclear cell (PBMC) fractions to investigate mt DNA content via Real-Time PCR in patients, not accounting platelets falsifying the mitochondrial (mt)DNA:nuclear (n)DNA-ratio. In this study we suggest a procedure to eliminate disturbing platelets totally. 8 healthy controls (G1), 6 therapy-naive HIV-infected patients (G2) and 9 HIV-infected patients under HAART (G3) were examined for mtDNA:nDNA-ratio using Real-Time PCR technology. Different blood collection and/or PBMC isolation strategies were analysed for variances of outcome at examinations of the same blood donor. Using DNA prepared of whole blood specimens, mtDNA:nDNA-ratios showed no differences in all investigated groups (G1, G2, G3). Comparing mtDNA:nDNA-ratios of platelet-depleted PBMC fractions of G1 with G2 revealed a reduction of 22% (p = 0.128) and a steeper reduction of 40% (p = 0.0036) comparing specimens of G1 with G3. Scrutinising differently processed specimens within the groups themselves, in G2 whole blood versus platelet-containing PBMC specimens showed a difference in mtDNA:nDNA ratios of 26% (p = 0.0406), whereas a comparison of whole blood versus platelet free PBMC specimens led to a comparatively more distinct reduction of 35% (p = 0.0089). The same effect was seen in G3, where whole blood versus platelet containing PBMC specimens revealed a reduction of 32% (p = 0.01) and whole blood versus platelet-free PBMC specimens showed a 42% (p = 0.0011) decrease. Furthermore analysing each single patient in relation to the different methods, a minor fluctuation margin could be found using platelet-free PBMC specimens for Real-Time PCR. Using platelet-free PBMCs for mt DNA content detection, a correlation of low mtDNA:nDNA-ratios to clinical signs, like elevated lactate levels or lipodystrophy, could be observed. Light-microscopic evaluation for platelets, comparing platelet-containing PBMC fractions versus platelet-depleted PBMC fractions reinforced the Real-Time PCR results. Our data demonstrate that the first step of the blood sample collection/preparation is critical for valid illustration of mt DNA content in HIV-infected patients using ultra-sensitive Real-Time PCR technology. The use of serum tubes for blood collection is an easy and low-cost alternative to expensive cell sorting for elimination of disturbing platelets. Using platelet-free PBMC fractions for measurement mt DNA content could be a surrogate marker for clinical signs mediated by HAART. PMID- 15337627 TI - n6/n3 hypothesis and allergies: biologically plausible, but not confirmed. AB - The dietary intake of certain fatty acids might contribute to the development of atopic diseases like allergic rhinitis and asthma. We investigated the association between the ratio of n-6 and n-3 fatty acids in serum phospholipids and hay fever or allergic sensitisation in adults. Data from a population based cross-sectional study on respiratory health including measurement of fatty acids in serum phospholipids of 740 adults between 20 and 64 years of age were analysed. We could not find any significant association between n6/n3-ratio of fatty acids in serum phospholipids and hay fever or allergic sensitisation neither in the total population nor in the sub-population stratified by sex. Since no previous study on fatty acid intake confirmed the n6/n3-ratio hypothesis and this study did not find any association between the n6/n3-ratio in serum phospholipids and hay fever or allergic sensitization, we conclude that the n6/n3 ratio hypothesis is not confirmed although biological plausible. PMID- 15337628 TI - Comparison of in vitro-activity of commonly used topical glucocorticoids on cytokine- and phospholipase inhibition. AB - BACKGROUND AND OBJECTIVES: Topical glucocorticoids (GCs) are potent inhibitors of cellular inflammatory mediator production. Differences in receptor binding activities are believed to correlate with inhibition of mediator release and anti inflammatory efficacy in vivo. To further assess this hypothesis we compared in cultured human monocytes the inhibitory activity of classic synthetic GCs on leukotriene B4 (LTB4), prostaglandin E2 (PGE2), interleukin 1 beta (IL-1beta) and c-phospholipase A 2 activity (cPLA2). METHODS: Normal human monocytes (10(5) /ml) were tested for 20 hrs with increasing concentrations (range 10(-12) -10(-5) M) of triamcinolone acetonide (TAA) compared to beclomethasone dipropionate (BDP), budesonide (BUD), dexamethasone (DEX), or the ethanol diluent together with 10 microg/ml of lipopolysaccharide (LPS). Mediator production and spontaneous cPLA subset 2-activity was determined by direct enzyme immunoassay methods. RESULTS: TAA at therapeutically relevant concentration (10(-8) M) inhibited significantly (p<0.01, n = 9) mediator production of TNF-alpha > IL-1beta > TxB2 > LTB subset 4 in a dose dependent manner by 75%, 65%, 41%, and 33%. IL-1beta inhibition at 10( 8) M by TAA (65%)> BDP (52%)> BUD (47%) was not different (ANOVA, p>0.2). Also spontaneous cPLA2-activity at 10(-8) M was inhibited to a similar degree (ANOVA, p> 0.6) by BUD (17.3%) > TAA (11.4%) > BDP (8.6%). In the same culture conditions spontaneous PGE2-secretion was inhibited by BDP (28.8%) > BUD (24.2%) > TAA (11.4%) with no significant effect for TAA. CONCLUSION: Clinically well established GCs have a similar inhibitory capacity on monocyte cytokine production and surprisingly only weak effects on AA-metabolism. Small receptor binding activity may account for the lack of cytokine inhibition by subtherapeutic (<10(-8) M) airway concentrations of TAA and BDP. Partial mediator inhibition by GCs at therapeutically known airway concentrations may be relevant to control bursts of airway inflammation during acute exacerbation but unfavourable to effectively delay progression of chronic airway inflammation. PMID- 15337629 TI - Improvement of the quality of diabetes control and decrease in the concentrations of AGE-products in patients with type 1 and insulin-treated type 2 diabetes mellitus: results from a 10 year-prospective, population-based survey on the quality of diabetes care in Germany (JEVIN). AB - BACKGROUND: Advanced glycation end (AGE)-products are a complex group of compounds that have been implicated in diabetes related long-term complications. Up to the present only few data exist about serum levels of the AGE-proteins N epsilon-Carboxymethyllysine (CML) and pentosidine in patients with diabetes mellitus. PATIENTS AND METHODS: In the present 10-year, population-based trial of a selection-free cohort of patients with insulin-treated diabetes mellitus, serum CML and pentosidine levels were examined in correlation to the patients' quality of diabetes control and the prevalence of diabetes related long-term complications. RESULTS: Following the reunification of Germany in 1989 the health care system was decentralised. Up to 1994/95 the relative HbA1c (HbA1c/mean normal) of patients with type 1 diabetes increased (1.65 +/- 0.35 versus 1.52 +/- 0.31, p = 0.002). For patients with type 2 diabetes it remained constant (1.75 +/ 0.4 versus 1.78 +/- 0.31, p = 0.669). During the following period (from 1994/95 to 1999/2000) specialised diabetes care, structured treatment and teaching programmes (TTP), intensified insulin therapy and blood glucose self-monitoring for all patients were broadly implemented. This was accompanied by a substantial improvement in the relative HbA1c of both, patients with type 1 (1.48 +/- 0.3, p<0.0001), and insulin-treated type 2 diabetes mellitus (1.47 +/- 0.25, p<0.0001). During the same period the mean concentration of the AGE-product CML in the sera of patients with type 1 and insulin-treated type 2 diabetes decreased (type 1: 1994/95: 1158.1 +/- 410.0 ng/ml versus 1999/2000: 938.5 +/- 422.4 ng/ml, p<0.0001, type 2: 1994/94: 1244.7 +/- 1231.3 ng/ml versus 1999/2000: 970.9 +/- 458.6 ng/ml, p = 0.007). For pentosidine the same tendency was found for patients with type 1 diabetes (1994/95: 253.6 +/- 280.7 pmol/ml versus 1999/2000: 148.2 +/ 91.4 pmol/ml, p<0.0001). For patients with type 1 diabetes there was a positive correlation between the relative HbA1c-value calculated over the total follow-up period of 10 years and the CML-concentration in 1999/2000 (r = 0.405, p = 0.017). In 1999/2000 a reduced creatinine clearance ( 0.05). Protein synthesis increase of the epithelial tumor cells (0.581 +/- 0.013 mg/ml, control) was observed with the conventional gutta-percha points (0.688 +/- 0.078 mg/ml) and with both gutta percha points containing different calcium hydroxide-based formulations (0.776 +/ 0.115 and 0.7 +/- 0.047 mg/ ml). CONCLUSIONS: Under the conditions of this study, chlorhexidine containing gutta-percha points showed the highest effect on cell growth inhibition. No significant differences were observed between the tested material and the two different cell culture types. PMID- 15337636 TI - Anthroposophic therapies in chronic disease: the Anthroposophic Medicine Outcomes Study (AMOS). AB - CONTEXT: Anthroposophic medicine (AM) is used worldwide for chronic diseases. OBJECTIVE: To study clinical outcomes and costs in patients treated with AM therapies for chronic conditions. DESIGN: Prospective cohort study. SETTING: 141 medical practices in Germany providing AM treatment. PARTICIPANTS AND INTERVENTIONS: 898 outpatients aged 1-75 years referred to AM therapies (art, eurythmy or rhythmical massage, n = 665) or starting AM medical treatment (counselling, medicines, n = 233). MAIN OUTCOME MEASURES: Disease severity assessed independently by physician (Disease Score) and patient (Symptom Score), and health-related quality of life (SF-36, KINDL, KITA) after 3, 6, 12, 18, and 24 months; health costs in pre-study year and first study year. RESULTS: Most common indications were mental disorders (32.0%), and musculoskeletal disorders (18.9%). Disease duration at baseline was median 3.0 years (interquartile range = i.q.r. 1.0-8.5, mean 6.5 +/- 8.4 years). Median number of AM therapy sessions was 12 (i.q.r. 10-20), median therapy duration was 120 days (i.q.r. 81-195). From baseline to 6-month follow-up, Disease Score (0-10) improved from 6.40 +/- 1.76 to 3.43 +/- 2.23 (p < 0.001), Symptom Score (0-10) improved from 5.89 +/- 1.75 to 3.35 +/- 2.09 (p < 0.001). In adults, SF-36 Physical Component Summary improved from mean 43.34 +/- 10.58 at baseline to 47.44 +/- 10.32 after 6 months (p < 0.001), SF-36 Mental Component Summary improved from 38.83 +/- 12.45 to 44.93 +/- 10.92 (p < 0.001). Similar HRQoL improvements were observed in children (KINDL, KITA). All improvements remained stable until 24-month follow-up. Adverse effects from AM therapies occurred in 2.7% (19/712) of patients. Three (0.5%) patients stopped therapy due to adverse effects. Health costs were 3,637 Euro per patient in the pre-study year and 3,484 Euro in the first study year, a decrease of 152 Euro (4.2%) per patient. CONCLUSION: Anthroposophic therapies were associated with long-term reduction of chronic disease symptoms, improvement of health related quality of life, and health cost reduction. PMID- 15337637 TI - Genotype-phenotype analysis in early-onset Alzheimer's disease due to presenilin 1 mutations at codon 139. AB - Mutations in the presenilin-1 (PS-1) gene are the main cause of autosomal dominant early onset Alzheimer's disease (EOAD) and show a high penetrance of symptoms. There are more than 100 mutations in the PS-1 gene. Among them are at present four different missense mutations known at position 139 on exon 5. Lack of genotyping in other family members may lead to the suggestion of sporadic cases. We present the case of a 46-year old German female with EOAD. Cognitive decline started at the age of 32, while myoclonic and tonic-clonic jerks occurred later. Disease symptoms were present in three generations of her family. Genetic analysis revealed the M139V mutation on exon 5 of the PS-1 gene. We compared the clinical data of this family with seven previously reported families and two sporadic cases with mutations at the codon 139. The genotype-phenotype analysis showed marked intrafamilial homogeneity, but interfamilial heterogeneity in relation to the onset, duration, and progression of the disease. Onset and duration were not correlated to the amino acid exchanged. Another modifying genetic or environmental factor is probable. PMID- 15337638 TI - Different mechanisms of action of beta2-adrenergic receptor agonists: a comparison of reproterol, fenoterol and salbutamol on monocyte cyclic-AMP and leukotriene B4 production in vitro. AB - BACKGROUND: Beta2-adrenergic receptor agonists have several effects on airway function, most of which are mediated in a variety of cell types resulting in increased c-AMP-production and inhibition of inflammatory mediator production. However, their stimulating effects on cAMP-production became known to be inversed by increasing phosphodiesterase (PDE) activity and degradation of cAMP. Therefore, in this study we have evaluated the efficacy of reproterol, a dual acting beta2-adrenoceptor agonist and PDE-inhibitor, as compared to salbutamol and fenoterol with respect to production of cAMP and LTB4 in cultured monocytes. METHODS: Isolated human monocytes (10(5)/ml) were incubated (n = 9) in suspension with beta2-adrenoceptor agonists (10(-10) -10(-4) M) for 30 minutes with and without IBMX. Then, cAMP production was determined following treatment with Triton-X100. Production of LTB4 was measured following incubation of beta2 adrenoceptor agonists for 4 hrs in the presence of LPS (10 mg/ml). cAMP and LTB subset 4 were measured in culture supernatants by enzyme immunoassay. RESULTS: At 10(-5) M, production of cAMP was significantly stimulated by reproterol > fenoterol > salbutamol in a dose-dependent manner to an extent of *128%, *65%, 13% (*p<0.04) respectively. In contrast, LTB4-production was inhibited significantly to a similar degree by salbutamol and reproterol in a dose dependent manner by 59% and 49% (10(-5) M, p<0.03), respectively, with decreasing inhibition (15%) after fenoterol. Following co-incubation with IBMX, cAMP production only increased significantly (p<0.002) after fenoterol (+110%) compared to salbutamol (+29%) and reproterol (+50%) (ANOVA, p<0.001). CONCLUSION: These data suggest effects of the theophylline constituent of reproterol to inhibit adenylyl cyclase induced phosphodiesterase activity. The advantageous synergistic effects of reproterol on cAMP-production need to be further explored in trials. PMID- 15337639 TI - Schizophrenia genetics and dysbindin: a corner turned? PMID- 15337640 TI - Antidepressants for bipolar depression: a systematic review of randomized, controlled trials. AB - OBJECTIVE: This study reviewed the evidence from randomized, controlled trials on the efficacy and safety of antidepressants in the short-term treatment of bipolar depression. METHOD: The authors performed a systematic review and meta-analysis of randomized, controlled trials. They searched the Cochrane Collaboration Depression, Anxiety, and Neurosis Controlled Trials Register, incorporating results of searches of MEDLINE, EMBASE, CINAHL, PsycLIT, PSYNDEX, and LILACS. The main outcome measures were the proportion of patients who clinically responded to treatment and the rate of switching to mania. RESULTS: Twelve randomized trials were included, with a total of 1,088 randomly assigned patients. Five trials compared one or more antidepressants with placebo: 75% of these patients were receiving a concurrent mood stabilizer or an atypical antipsychotic. Antidepressants were more effective than placebo. Antidepressants did not induce more switching to mania (the event rate for antidepressants was 3.8% and for placebo, it was 4.7%). Six trials allowed comparison between two antidepressants. The rate of switching for tricyclic antidepressants was 10%, and for all other antidepressants combined, it was 3.2%. CONCLUSIONS: Antidepressants are effective in the short-term treatment of bipolar depression. The trial data do not suggest that switching is a common early complication of treatment with antidepressants. It may be prudent to use a selective serotonin reuptake inhibitor or a monoamine oxidase inhibitor rather than a tricyclic antidepressant as first-line treatment. Given the limited evidence, there is a compelling need for further studies with longer follow-up periods and careful definition and follow-up of emerging mania and partial remission. PMID- 15337641 TI - Maternal infanticide associated with mental illness: prevention and the promise of saved lives. AB - OBJECTIVE: Although maternal infanticide is a rare event, a high proportion of cases occurs in the context of postpartum mental illness. The author reviews historical, legislative, and contemporary psychiatric perspectives on infanticide and discusses ways in which the psychiatric community can improve prevention of infanticide and promote appropriate treatment of mentally ill women who commit infanticide. METHOD: The case of Texas v. Andrea Yates, involving a mother with mental illness who drowned her five children, is used to illustrate society's complicated reactions to infanticide in the context of postpartum mental illness. RESULTS: In the United States, the complexity of the response to infanticide is demonstrated by the judicial system's reaction to such cases. Whereas England's Infanticide Law provides probation and mandates psychiatric treatment for mothers with mental illness who commit infanticide, "killer mothers" may face the death penalty in the United States. Contemporary neuroscientific findings support the position that a woman with postpartum psychosis who commits infanticide needs treatment rather than punishment and that appropriate treatment will deter her from killing again. Psychiatrists have a vital role in recognizing the signs and symptoms of peripartum psychiatric disorders, particularly postpartum psychosis, and in early identification of and intervention with at-risk mothers. CONCLUSIONS: The absence of formal DSM-IV diagnostic criteria for postpartum psychiatric disorders promotes disparate treatment under the law. The psychiatric community should develop guidelines for the treatment of postpartum disorders, foster sharing of knowledge between psychiatry and the law, and do more to enlighten society about the effects of mental illness on thought and behavior so that decisions about the treatment and punishment of mentally ill persons will not be left exclusively in the hands of the judicial system. PMID- 15337642 TI - Protection of human subjects in intervention research for suicidal behavior. AB - OBJECTIVE: Three domains of ethical conduct outlined in the U.S. Food and Drug Administration's 1998 Belmont Report on protection of human research subjects respect for persons, beneficence, and justice-have posed specific dilemmas in the design of intervention studies for suicidal behavior. These issues include questions about suicidal patients' capacity to provide informed consent, the risk of some lethal outcomes, the possibility of imminent suicide risk associated with patients' right to discontinue the study treatment, and the need for a higher level of monitoring of suicidal patients. The authors examine these and other issues and discuss ways they can be addressed in research design. METHOD: To illustrate solutions to these bioethical tensions, the authors describe the design of a randomized, controlled trial of pharmacotherapy for suicidal behavior in bipolar disorder. RESULTS: Using surrogate outcome measures, allowing prescription of rescue medications, integrating psychosocial interventions, and providing close clinical monitoring enable researchers to conduct research on suicidal behavior while maximizing adherence to the ethical recommendations outlined in the Belmont Report. Alternative study designs, such as add-on trials, in which the study treatment or placebo is added on to known effective treatment, may also be used in research on suicidal behavior. CONCLUSIONS: It is possible to design a randomized, controlled trial that minimizes the risk of morbidity and mortality for suicidal patients with bipolar disorder, but deliberation is required to address the bioethical tensions that arise. PMID- 15337643 TI - Increased cortical neuronal density in schizophrenia. PMID- 15337644 TI - Comparing cognitive behavior therapy, interpersonal psychotherapy, and psychodynamic psychotherapy. PMID- 15337645 TI - Adolf Wolfli, 1864-1930. PMID- 15337646 TI - Prediction of antidepressant response to milnacipran by norepinephrine transporter gene polymorphisms. AB - OBJECTIVE: With a multitude of antidepressants available, predictors of response to different classes of antidepressants are of considerable interest. The purpose of the present study was to determine whether norepinephrine transporter gene (NET) and serotonin transporter gene (5-HTT) polymorphisms are associated with the antidepressant response to milnacipran, a dual serotonin/norepinephrine reuptake inhibitor. METHOD: Ninety-six Japanese patients with major depressive disorder were treated with milnacipran, 50-100 mg/day, for 6 weeks. Severity of depression was assessed with the Montgomery-Asberg Depression Rating Scale. Assessments were carried out at baseline and at 1, 2, 4, and 6 weeks of treatment. The method of polymerase chain reaction was used to determine allelic variants. RESULTS: Eighty patients completed the study. The presence of the T allele of the NET T-182C polymorphism was associated with a superior antidepressant response, whereas the A/A genotype of the NET G1287A polymorphism was associated with a slower onset of therapeutic response. In contrast, no influence of 5-HTT polymorphisms on the antidepressant response to milnacipran was detected. CONCLUSIONS: The results suggest that NET but not 5-HTT polymorphisms in part determine the antidepressant response to milnacipran. PMID- 15337647 TI - Genetic and environmental sources of covariation between generalized anxiety disorder and neuroticism. AB - OBJECTIVE: The authors examined the sources of covariation between generalized anxiety disorder and the personality trait of neuroticism. Because women have higher levels of neuroticism and twice the risk of lifetime generalized anxiety disorder of men, gender-specific effects were also explored. METHOD: Lifetime generalized anxiety disorder and neuroticism were assessed in more than 8,000 twins from male-male, female-female, and opposite-sex pairs through structured diagnostic interviews. Sex-limited Cholesky structural equation models were used to decompose the correlations between generalized anxiety disorder and neuroticism into genetic and environmental components, including sex-specific factors. RESULTS: Genetic correlations between generalized anxiety disorder and neuroticism were high and differed (nonsignificantly) between men and women (1.00 and 0.58, respectively). When nonsignificant gender differences were removed from the models, correlations between generalized anxiety disorder and neuroticism were estimated at 0.80 (95% confidence interval=0.52-1.00). The individual specific environmental correlation between generalized anxiety disorder and neuroticism was estimated at 0.20 for both genders. CONCLUSIONS: There is substantial overlap between the genetic factors that influence individual variation in neuroticism and those that increase liability for generalized anxiety disorder, irrespective of gender. The life experiences that increase vulnerability to generalized anxiety disorder, however, have only modest overlap with those that contribute to an individual's level of neuroticism. PMID- 15337648 TI - Psychiatric disorders among offspring of depressed mothers: associations with paternal psychopathology. AB - OBJECTIVE: The association between maternal depression and offspring dysfunction is well documented; however, little attention has been paid to psychopathology in the partners of these depressed mothers or to how paternal psychopathology might influence the relationship between maternal depression and offspring dysfunction. The purpose of this study was to explore whether major depression and/or antisocial behavior tended to occur more frequently among partners of depressed mothers (compared to partners of nondepressed mothers) and to examine how these paternal disorders related to offspring psychopathology. METHOD: Participants were drawn from the Minnesota Twin Family Study, a community-based study of twins and their parents. Depressed and nondepressed mothers, their partners (the biological fathers of the twins), and their 17-year-old offspring were included. Structured interviews were used to assess participants for the presence of major depression, conduct disorder, and adult antisocial behavior. RESULTS: Depressed mothers tended to partner with antisocial fathers. Depression in mothers and antisocial behavior in fathers were both significantly and independently associated with offspring depression and conduct disorder. No interactions of the parental diagnoses with each other or with the gender of the offspring were found. CONCLUSIONS: Many offspring of depressed mothers experience the additional risk of having an antisocial father. The implications of these findings for risk among the offspring of depressed mothers are discussed. PMID- 15337649 TI - Sex differences in functional connectivity in first-episode and chronic schizophrenia patients. AB - OBJECTIVE: There has been consistent evidence for a lower incidence and milder course of schizophrenia in women, yet there have been very few investigations of sex differences in brain function in this disorder. This study used a new high temporal-resolution measure of functional brain connectivity to test the prediction that female patients would show relatively greater inter- and intrahemispheric connectivity than male patients, particularly in the early stage of schizophrenia. METHOD: Forty patients with chronic schizophrenia (20 women and 20 men) and 24 patients with first-episode schizophrenia (12 women and 12 men) and their respective matched comparison groups completed a conventional auditory oddball task. Phase synchronous gamma (40 Hz) activity was extracted from EEG recording during the task and time-locked to the oddball (target) stimuli. RESULTS: Chronic schizophrenia subjects showed a reduction in global functional connectivity (lower gamma phase synchrony) relative to their matched healthy subjects. Unexpectedly, this reduction was most apparent in female patients. By contrast, while first-episode patients showed a general reduction in the speed of frontal connectivity, the speed of global connectivity was relatively faster in female patients. CONCLUSIONS: This is the first study to investigate sex differences in schizophrenia that used the functional connectivity measure of gamma phase synchrony. The results suggest that in female patients with schizophrenia, additional breakdowns in brain network connectivity may develop with illness chronicity. PMID- 15337650 TI - Middle and inferior temporal gyrus gray matter volume abnormalities in chronic schizophrenia: an MRI study. AB - OBJECTIVE: The middle temporal gyrus and inferior temporal gyrus subserve language and semantic memory processing, visual perception, and multimodal sensory integration. Functional deficits in these cognitive processes have been well documented in patients with schizophrenia. However, there have been few in vivo structural magnetic resonance imaging (MRI) studies of the middle temporal gyrus and inferior temporal gyrus in schizophrenia. METHOD: Middle temporal gyrus and inferior temporal gyrus gray matter volumes were measured in 23 male patients diagnosed with chronic schizophrenia and 28 healthy male subjects by using high spatial-resolution MRI. For comparison, superior temporal gyrus and fusiform gyrus gray matter volumes were also measured. Correlations between these four regions and clinical symptoms were also investigated. RESULTS: Relative to healthy subjects, the patients with chronic schizophrenia showed gray matter volume reductions in the left middle temporal gyrus (13% difference) and bilateral inferior temporal gyrus (10% difference in both hemispheres). In addition, the patients showed gray matter volume reductions in the left superior temporal gyrus (13% difference) and bilateral fusiform gyrus (10% difference in both hemispheres). More severe hallucinations were significantly correlated with smaller left hemisphere volumes in the superior temporal gyrus and middle temporal gyrus. CONCLUSIONS: These results suggest that patients with schizophrenia evince reduced gray matter volume in the left middle temporal gyrus and bilateral reductions in the inferior temporal gyrus. In conjunction with findings of left superior temporal gyrus reduction and bilateral fusiform gyrus reductions, these data suggest that schizophrenia may be characterized by left hemisphere-selective dorsal pathophysiology and bilateral ventral pathophysiology in temporal lobe gray matter. PMID- 15337651 TI - The entorhinal cortex in first-episode psychotic disorders: a structural magnetic resonance imaging study. AB - OBJECTIVE: Neuropathological findings regarding the entorhinal cortex in schizophrenia are conflicting. The authors used structural magnetic resonance imaging to examine the entorhinal cortex volumes of healthy subjects and medication-naive patients experiencing their first episode of psychotic illness. METHOD: The study included 33 patients with schizophrenia and related disorders, 11 patients with nonschizophrenic disorders, and 43 matched healthy subjects. All subjects were rated on the Scale for the Assessment of Positive Symptoms and the Scale for the Assessment of Negative Symptoms, and volumetric measurements of the entorhinal cortex were obtained for all subjects. The authors examined differences across the groups as well as clinical correlations of entorhinal cortex volumes adjusted for intracranial volume. RESULTS: A significant diagnosis effect was seen in the left entorhinal cortex: patients with schizophrenia and related disorders and patients with nonschizophrenic psychotic disorders had smaller left entorhinal cortex volumes than healthy subjects. The mean entorhinal cortex volume of patients with schizophrenic disorders did not differ from that of patients with nonschizophrenic psychotic disorders. In patients with schizophrenic disorders, the entorhinal cortex volume positively correlated with severity of delusions. The mean entorhinal cortex volume of patients with nondelusional psychotic disorders was significantly smaller than that of patients with delusional psychotic disorders and healthy subjects. CONCLUSIONS: Smaller entorhinal cortex volume in first-episode, neuroleptic-naive psychotic disorders may not be a confound of the effects of illness chronicity or antipsychotic treatment. Entorhinal cortex pathology appears to have a significant association with positive symptoms, specifically delusions. The impairment of functions in which the entorhinal cortex participates-such as novelty detection, associative learning, and processing episodic, recognition, and autobiographical memory-could be responsible for its association with psychotic disorders and delusions. PMID- 15337652 TI - Equivalent occupancy of dopamine D1 and D2 receptors with clozapine: differentiation from other atypical antipsychotics. AB - OBJECTIVE: Clozapine, the prototype of atypical antipsychotics, remains unique in its efficacy in the treatment of refractory schizophrenia. Its affinity for dopamine D(4) receptors, serotonin 5-HT(2A) receptor antagonism, effects on the noradrenergic system, and its relatively moderate occupancy of D(2) receptors are unlikely to be the critical mechanism underlying its efficacy. In an attempt to elucidate the molecular/synaptic mechanism underlying clozapine's distinctiveness in refractory schizophrenia, the authors studied the in vivo D(1) and D(2) receptor profile of clozapine compared with other atypical antipsychotics. METHOD: Positron emission tomography with the radioligands [(11)C]SCH23390 and [(11)C]raclopride was used to investigate D(1) and D(2) receptor occupancy in vivo in 25 schizophrenia patients receiving atypical antipsychotic treatment with clozapine, olanzapine, quetiapine, or risperidone. RESULTS: Mean striatal D(1) occupancies ranged from 55% with clozapine to 12% with quetiapine (rank order: clozapine > olanzapine > risperidone > quetiapine). The striatal D(2) occupancy ranged from 81% with risperidone to 30% with quetiapine (rank order: risperidone > olanzapine > clozapine > quetiapine). The ratio of striatal D(1)/D(2) occupancy was significantly higher for clozapine (0.88) relative to olanzapine (0.54), quetiapine (0.41), or risperidone (0.31). CONCLUSIONS: Among the atypical antipsychotics, clozapine appears to have a simultaneous and equivalent occupancy of dopamine D(1) and D(2) receptors. Whether its effect on D(1) receptors represents agonism or antagonism is not yet clear, as this issue is still unresolved in the preclinical arena. This distinctive effect on D(1)/D(2) receptors may be responsible for clozapine's unique effectiveness in patients with schizophrenia refractory to other typical and atypical antipsychotics. PMID- 15337653 TI - Prevalence and predictors of depression treatment in an international primary care study. AB - OBJECTIVE: The purpose of the study was to evaluate the prevalence and predictors of depression treatment in a diverse cross-national sample of primary care patients. METHOD: At primary care facilities in six countries (Spain, Israel, Australia, Brazil, Russia, and the United States), a two-stage screening process was used to identify 1,117 patients with current depressive disorder. At baseline, all patients completed a structured diagnostic interview as well as measures of anxiety symptoms, alcohol use, chronic comorbid physical conditions, and perceived barriers to treatment. Primary care physicians were advised if the research interview indicated a probable depressive disorder in their patients. Three and 9 months later, participants reported all health services (including specialty mental health care and antidepressant medication) used in the preceding 3 months. RESULTS: Across the six sites, the proportion of patients receiving any antidepressant pharmacotherapy ranged from a high of 38% in Seattle to a low of 0% in St. Petersburg; the proportion receiving any specialty mental health care varied from a high of 29% in Melbourne to a low of 3% in St. Petersburg. Patient characteristics were not consistently associated with receipt of either pharmacotherapy or specialty mental health care. Out-of-pocket cost was the most commonly reported barrier to treatment for depression; the percentage of patients who reported this barrier ranged from 24% in Barcelona to 75% in St. Petersburg. CONCLUSIONS: Depression screening and physician notification are not sufficient to prompt adequate treatment for depression. The probability of treatment may be more influenced by characteristics of health care systems than by the clinical characteristics of individual patients. Financial barriers may be more important than stigma as impediments to appropriate care. PMID- 15337654 TI - Racial disparity in the use of ECT for affective disorders. AB - OBJECTIVE: Published reports indicate that African Americans are underrepresented among patients treated with ECT. The reason for this disparity in practice has not been determined. This study addressed this question by using existing data on a large series of patients treated with ECT at a single academic medical center. METHOD: The hospital's administrative databases were used to select Caucasian and African American patients with a diagnosis of major affective disorder treated over the period from November 1993 to March 2002. Independent variables were age, sex, treatment unit, readmission within 30 days, type of insurance, and geographic zone of residence. The dependent variable was likelihood of being treated with ECT, computed for each race group. RESULTS: Caucasians were more likely than African Americans to be treated with ECT (odds ratio=4.71; 95% confidence interval [CI]=3.77-5.90). None of the variables examined provided an explanation for this disparity. When all of the variables were controlled simultaneously, the likelihood of being treated with ECT remained significantly higher for Caucasians than for African Americans (odds ratio=2.48; 95% CI=1.89 3.25). CONCLUSIONS: The racial disparity in the use of ECT cannot be explained on the basis of the variables studied. It is not an artifact of the age of the patient population, nor can it be explained on the basis of insurance coverage, the social class of the patients, or their illnesses' being more treatment resistant. The authors propose several other hypotheses, including explanations relating to clinical presentation, differential response to other treatments, differences in patients' willingness to consent, and physicians' behavior, that could be explored by using other methods. PMID- 15337655 TI - Efficacy of sertraline in a 12-week trial for generalized anxiety disorder. AB - OBJECTIVE: Sertraline's efficacy and tolerability in treating generalized anxiety disorder were evaluated. METHOD: Adult outpatients with DSM-IV generalized anxiety disorder and a total score of 18 or higher on the Hamilton Anxiety Rating Scale were eligible. After a 1-week single-blind placebo lead-in, patients were randomly assigned to 12 weeks of double-blind treatment with placebo (N=188, mean baseline anxiety score=25) or flexible doses (50-150 mg/day) of sertraline (N=182, mean anxiety score=25). The primary outcome measure was baseline-to endpoint change in the Hamilton anxiety scale total score. A secondary efficacy measure was the Clinical Global Impression (CGI) improvement score; response was defined as a score of 2 or less. RESULTS: Sertraline patients had significantly greater improvement than placebo patients on all efficacy measures at week 4. Analysis of covariance of the intent-to-treat group at endpoint (with the last observation carried forward) showed a significant difference in the decrease from baseline of the least-square mean total score on the Hamilton anxiety scale between sertraline (mean=11.7) and placebo (mean=8.0). Significantly greater endpoint improvement with sertraline than placebo was obtained for mean scores on the Hamilton anxiety scale psychic factor (6.7 versus 4.1) and somatic factor (5.0 versus 3.9). The rate of responders, based on CGI improvement and last observation carried forward, was significantly higher for sertraline (63%) than placebo (37%). Sertraline was well tolerated; 8% of patients versus 10% for placebo dropped out because of adverse events. CONCLUSIONS: Sertraline appears to be efficacious and well tolerated in the treatment of generalized anxiety disorder. PMID- 15337656 TI - Response inhibition in adolescents diagnosed with attention deficit hyperactivity disorder during childhood: an event-related FMRI study. AB - OBJECTIVE: Frontostriatal neural abnormalities have been implicated in the response inhibition impairments that are characteristic of attention deficit hyperactivity disorder (ADHD). However, reports of such abnormalities in adolescents are inconsistent. The present study used behavioral and functional neuroimaging techniques to examine inhibitory control processes in adolescents who had been diagnosed with ADHD during childhood. METHOD: The authors used functional magnetic resonance imaging (fMRI) during performance of a Go/No-Go task to scan 10 male adolescents who were diagnosed with DSM-III-R ADHD when they were 7 to 11 years old and nine age-, sex-, and IQ-matched comparison subjects with no history of ADHD. Response inhibition was tested by contrasting neural activation during No-Go trials with that during Go trials. RESULTS: The inhibition of a prepotent tendency to respond produced markedly greater activation of the left anterior cingulate gyrus, bilateral frontopolar regions, bilateral ventrolateral prefrontal cortex, and left medial frontal gyrus in the adolescents with childhood ADHD than in the adolescents with no history of ADHD. Activity in the first two regions was inversely related to task performance across the study group. CONCLUSIONS: Compared with adolescents who had no history of ADHD, adolescents who were diagnosed with ADHD during childhood exhibited enhanced responses during inhibition in ventrolateral prefrontal cortical areas that subserve response inhibition, as well as in anterior cingulate and frontopolar regions implicated in other executive functions. PMID- 15337657 TI - Testing psychodynamic psychotherapy skills among psychiatric residents: the psychodynamic psychotherapy competency test. AB - OBJECTIVE: Training in psychodynamic psychotherapy remains a core requirement of psychiatric residency training programs, yet no standard measures of competency exist to document residents' knowledge and skills in this area. To address this issue, the authors developed a written test of applied knowledge of psychodynamic psychotherapy technique and theory, the Psychodynamic Psychotherapy Competency Test. Their goal in this article was to evaluate the validity of this test. METHOD: The test was given to a group of 36 psychoanalytic experts and 206 residents in their second, third, and fourth psychiatric postgraduate years from 10 programs located in different parts of the United States. Program directors provided information on the number of hours of psychodynamic didactic teaching, supervision, and resident-conducted psychodynamic psychotherapy and rated the psychodynamic psychotherapy skills of residents in their fourth postgraduate year on the basis of cumulative supervisor reports. RESULTS: There were significant differences in test performance between residents and faculty experts and between residents in their second and fourth postgraduate years: more advanced residents and experts had progressively better scores. The mean scores of fourth-year residents in different programs differed significantly, but the scores of second year residents did not. Higher test scores were positively associated with both number of hours of resident-conducted psychotherapy and number of hours of supervision. Among fourth-year residents, test scores correlated significantly with program director evaluations. CONCLUSIONS: This initial study supports the validity of the Psychodynamic Psychotherapy Competency Test as well as the feasibility of testing psychotherapy skills in a standardized fashion. PMID- 15337658 TI - Long-term changes in defense styles with psychodynamic psychotherapy for depressive, anxiety, and personality disorders. AB - OBJECTIVE: This study examined 1) whether patients with chronic and recurrent anxiety and depressive disorders and/or personality disorders demonstrate improvement in their defense styles with long-term dynamic psychotherapy and 2) what the relationship is between defense style change and symptomatic change. METHOD: Measures of defense (Defense Style Questionnaire) and symptoms and functioning were administered at regular intervals over the course of 3-5 years to adults who entered a naturalistic study of long-term psychodynamic psychotherapy. With hierarchical linear regression, the relative contributions of change in variables on the Defense Style Questionnaire to change in other outcome variables were calculated. RESULTS: Those with high initial scores on the maladaptive and self-sacrificing defense styles improved, with effect sizes of 0.80 and 0.67, while overall defensive functioning improved, with an effect size of 0.43. The effect size of the change in score on the Global Assessment of Functioning scale was 0.82. Depressed subjects improved their scores significantly on the Hamilton Depression Rating Scale, and there was a significant improvement in distress, as measured by the SCL-90-R. Changes in score on the Defense Style Questionnaire added substantially to the prediction of variance in these three outcomes above their initial levels. A higher level of defensive functioning also predicted a better self-reported therapeutic alliance. CONCLUSIONS: Defense styles became more adaptive and symptoms improved over time in patients who started with scores in the clinical range. Change in defense style predicts symptomatic change, but causation has not been established. PMID- 15337659 TI - Health correlates of recreational gambling in older adults. AB - OBJECTIVE: Prior studies have found high rates of alcohol use and abuse/dependence, depression, bankruptcy, and incarceration associated with recreational gambling. Despite growing rates of recreational gambling in older adults, little is known regarding its health correlates in this age group. The objective of this study was to identify health and well-being correlates of past year recreational gambling in adults age 65 years and older, compared to adults age 18-64 years. METHOD: The Gambling Impact and Behavior Study surveyed by telephone a nationally representative sample of 2,417 adults. Multivariate analyses were used to compare past-year recreational gamblers and nongamblers in the older and younger age groups on measures of alcohol use and abuse/dependence, substance abuse/dependence, depression, mental health treatment, subjective general health, incarceration, and bankruptcy. Additional analyses compared the gambling patterns in older and younger adult past-year recreational gamblers. RESULTS: After the effects of sociodemographic factors were controlled, older adult past-year recreational gamblers were more likely to report past-year alcohol use and better health than were older nongamblers. Multivariate analyses investigating interactions of gambling and age found that higher rates of good to excellent subjective general health in recreational gamblers were mainly attributable to the older age group. Older adult gamblers were more likely than younger adult gamblers to begin gambling after age 18 years, to gamble more frequently, and to report a larger maximum win. CONCLUSIONS: Recreational gambling patterns of older adults differ from those of younger adults. In contrast to findings in younger adults, recreational gambling in older adults is not associated with negative measures of health and well-being. PMID- 15337660 TI - A new, empirically established hypochondriasis diagnosis. AB - OBJECTIVE: The narrow ICD-10 and DSM-IV definition of hypochondriasis makes it rarely used yet does not prevent extensive diagnosis overlap. This study identified a distinct hypochondriasis symptom cluster and defined diagnostic criteria. METHOD: Consecutive patients (N=1,785) consulting primary care physicians for new illness were screened for somatization, anxiety, depression, and alcohol abuse. A stratified subgroup of 701 patients were interviewed with the Schedules for Clinical Assessment in Neuropsychiatry and questions addressing common hypochondriasis symptoms. Symptom patterns were analyzed by latent class analysis. RESULTS: Patients fell into three classes based on six symptoms: preoccupation with the idea of harboring an illness or with bodily function, rumination about illness, suggestibility, unrealistic fear of infection, fascination with medical information, and fear of prescribed medication. All symptoms, particularly rumination, were frequent in one of the classes. Classification allowed definition of new diagnostic criteria for hypochondriasis and division of the cases into "mild" and "severe." The weighted prevalence of severe cases was 9.5% versus 5.8% for DSM-IV hypochondriasis. Compared with DSM IV hypochondriasis, this approach produced less overlap with other somatoform disorders, similar overlap with nonsomatoform psychiatric disorders, and similar assessments by primary care physicians. Severe cases of the new hypochondriasis lasted 2 or more years in 54.3% of the subjects and 1 month or less in 27.2%. CONCLUSIONS: These results suggest that rumination about illness plus at least one of five other symptoms form a distinct diagnostic entity performing better than the current DSM-IV hypochondriasis diagnosis. However, these criteria are preliminary, awaiting cross-validation in other subject groups. PMID- 15337661 TI - Safety reporting in randomized trials of mental health interventions. AB - OBJECTIVE: The authors aimed to evaluate the adequacy of the reporting of safety information in publications of randomized trials of mental-health-related interventions. METHOD: The authors randomly selected 200 entries from the PsiTri registry of mental-health-related controlled trials. This yielded 142 randomized trials that were analyzed for adequacy and relative emphasis of their content on safety issues. They examined drug trials as well as trials of other types of interventions. RESULTS: Across the 142 eligible trials, 103 involved drugs. Twenty-five of the 142 trials had at least 100 randomly chosen subjects and at least 50 subjects in a study arm. Among drug trials, only 21.4% had adequate reporting of clinical adverse events, and only 16.5% had adequate reporting of laboratory-determined toxicity, while 32.0% reported both the numbers and the reasons for withdrawals due to toxicity in each arm. On average, drug trials devoted 1/10 of a page in their results sections to safety, and 58.3% devoted more space to the names and affiliations of authors than to safety. None of the trials of nondrug interventions had adequate or even partially adequate reporting of either clinical adverse events or laboratory-determined toxicity. In multivariate modeling, long-term trials and trials conducted in the United States devoted even less space to safety, while schizophrenia trials devoted more space to safety than did trials in other areas. CONCLUSIONS: Safety reporting is largely neglected across trials of mental-health-related interventions, thus hindering the assessment of risk-benefit ratios for rational decision making in mental health care. PMID- 15337662 TI - Linkage disequilibrium of the brain-derived neurotrophic factor Val66Met polymorphism in children with a prepubertal and early adolescent bipolar disorder phenotype. AB - OBJECTIVE: Transmission of the brain-derived neurotrophic factor (BDNF) Val66 allele in children with a prepubertal and early adolescent bipolar disorder phenotype was examined. METHOD: The prepubertal and early adolescent bipolar disorder phenotype was defined as current DSM-IV bipolar I disorder (manic or mixed phase) with at least one cardinal mania criterion (i.e., euphoria and/or grandiosity) to ensure differentiation from attention deficit hyperactivity disorder. Probands (mean age=10.7 years, SD=2.7) were obtained by consecutive new case ascertainment from designated pediatric and psychiatric venues. Parents and probands were interviewed separately by research nurses who were blind to the probands' diagnoses. Genotyping was done with TaqMan Assay-on-Demand. Analysis was done with the Family Based Association Test program. RESULTS: There were 53 complete, independent trios. The BDNF Val66 allele was preferentially transmitted (Family Based Association Test: chi(2)=6.0, df=1, p=0.014). CONCLUSIONS: This finding in child bipolar disorder is consistent with data for adults with bipolar disorder that show preferential transmission of the Val66 allele. PMID- 15337663 TI - Effects of a functional COMT polymorphism on prefrontal cognitive function in patients with 22q11.2 deletion syndrome. AB - OBJECTIVE: The 22q11.2 deletion syndrome (DiGeorge/velocardiofacial syndrome) is associated with attentional problems and executive dysfunction, and is one of the highest known risk factors for schizophrenia. These behavioral manifestations of 22q11.2 deletion syndrome could result from haploinsufficiency of the catechol O methyltransferase (COMT) gene, located within the 22q11 region. The goal of the present study was to examine COMT genotype as a predictor of prefrontal cognitive function in patients with 22q11.2 deletion syndrome. METHOD: Patients with confirmed 22q11.2 deletions (N=44) underwent neurocognitive testing following Val(158)Met genotyping (Met hemizygous: N=16; Val hemizygous: N=28). RESULTS: Analyses of covariance revealed that Met-hemizygous patients performed significantly better on a composite measure of executive function (comprising set shifting, verbal fluency, attention, and working memory) than did Val-hemizygous patients. CONCLUSIONS: These data are consistent with those of previous studies in normal individuals, suggesting that a functional genetic polymorphism in the 22q11 region may influence prefrontal cognition in individuals with COMT haploinsufficiency. PMID- 15337664 TI - Association between catechol-O-methyltransferase and phobic anxiety. AB - OBJECTIVE: The authors assessed the association between the catechol-O methyltransferase (COMT) Val158Met polymorphism and scores on the phobic anxiety scale of the Crown-Crisp Experimental Index. METHOD: A total of 1,234 women completed the Crown-Crisp Experimental Index phobic anxiety scale and were genotyped for the COMT polymorphism. The authors used unconditional logistic regression to compute odds ratios and 95% confidence intervals (CIs) to evaluate the association between the COMT genotype and phobic anxiety. RESULTS: The mean scores for the three genotypes were statistically significantly different. Compared to the COMT Met/Met genotype, the age-adjusted odds ratio for scoring >/=6 compared to scoring 0 or 1 were 1.15 (95% CI=0.71-1.85) and 1.99 (95% CI=1.17-3.40) for the COMT Val/Met and COMT Val/Val genotypes, respectively; a significant gene dosage effect was observed. CONCLUSIONS: Our results suggest that the functional COMT polymorphism is associated with the development of phobic anxiety. PMID- 15337665 TI - Folate, homocysteine, and negative symptoms in schizophrenia. AB - OBJECTIVE: Because glutamate carboxypeptidase II (GCPII) regulates both folate absorption and activation of N-methyl-d-aspartic acid receptors, the authors examined relationships between serum folate concentrations and clinical symptoms in schizophrenia patients. METHOD: For 91 outpatients with schizophrenia, clinical assessments were performed and serum folate, homocysteine, B(12), glycine, and serine concentrations were measured. RESULTS: Serum folate concentrations were significantly lower than in a representative sample from the Framingham Offspring Study. Folate concentration correlated inversely with the Scale for Assessment of Negative Symptoms total score and was lower in patients with the deficit syndrome than in nondeficit patients. Homocysteine concentration correlated with the severity of extrapyramidal symptoms. CONCLUSIONS: These findings could reflect several possible mechanisms, including low dietary intake of folate, low GCPII activity, cigarette smoking, and the involvement of folate in the synthesis of neurotransmitters. Additional studies are needed to clarify these findings. PMID- 15337666 TI - Incidence of newly diagnosed diabetes attributable to atypical antipsychotic medications. AB - OBJECTIVE: The purpose of the study was to determine the proportion of patients with schizophrenia with a stable regimen of antipsychotic monotherapy who developed diabetes or were hospitalized for ketoacidosis. METHOD: Patients with schizophrenia for whom a stable regimen of antipsychotic monotherapy was consistently prescribed during any 3-month period between June 1999 and September 2000 and who had no diabetes were followed through September 2001 by using administrative data from the Department of Veterans Affairs. Cox proportional hazards models were developed to identify the characteristics associated with newly diagnosed diabetes and ketoacidosis. RESULTS: Of the 56,849 patients identified, 4,132 (7.3%) developed diabetes and 88 (0.2%) were hospitalized for ketoacidosis. Diabetes risk was highest for clozapine (hazard ratio=1.57) and olanzapine (hazard ratio=1.15); the diabetes risks for quetiapine (hazard ratio=1.20) and risperidone (hazard ratio=1.01) were not significantly different from that for conventional antipsychotics. The attributable risks of diabetes mellitus associated with atypical antipsychotics were small, ranging from 0.05% (risperidone) to 2.03% (clozapine). CONCLUSIONS: Although clozapine and olanzapine have greater diabetes risk, the attributable risk of diabetes mellitus with atypical antipsychotics is small. PMID- 15337667 TI - Insight and its relationship to violent behavior in patients with schizophrenia. AB - OBJECTIVE: Lack of insight affects the management of schizophrenia. The interrelationship between lack of insight and illness attributes in patients with schizophrenia who commit violent acts is important and underresearched. METHOD: One hundred fifteen violent patients with schizophrenia in a jail or court psychiatric clinic were evaluated on measures of symptoms, illness severity, insight into illness, and the legal consequences of their illness ("forensic insight"). A sample of nonviolent patients served as a comparison group. RESULTS: Compared with the nonviolent cohort, violent patients were more symptomatic, had poorer functioning, and had a more prominent lack of insight. Deficits of insight into illness coexisted with a lack of forensic insight, which was also associated with psychosis. CONCLUSIONS: Patients with schizophrenia who commit violent acts have insight deficits, including lack of awareness of the legal implications of their behavior. Targeted interventions to improve insight and treatment compliance in this population are warranted. PMID- 15337668 TI - ECT in delusional depression with multiple sclerosis. PMID- 15337669 TI - Excretion of quetiapine in breast milk. PMID- 15337670 TI - Modafinil augmentation of phenelzine for residual fatigue in dysthymia. PMID- 15337671 TI - Neuroacanthocytosis. PMID- 15337672 TI - Practice guidelines and combining atypical antipsychotics. PMID- 15337673 TI - Intranasal quetiapine abuse. PMID- 15337674 TI - Atomoxetine and nonresponders to stimulants. PMID- 15337675 TI - Autism, movement, and facial processing. PMID- 15337676 TI - Religion, spirituality, and mysticism. PMID- 15337677 TI - Conflict-of-interest charge. PMID- 15337678 TI - Potential for misuse of sedatives. PMID- 15337679 TI - Heteromodal association cortex in schizophrenia. PMID- 15337680 TI - Psychodynamic therapy and the journal. PMID- 15337681 TI - Importance of learning in the response of ewes to male odor. AB - Exposure of anestrous ewes to a ram or its odor results in the activation of the luteinizing hormone (LH) secretion leading to reinstatement of cyclicity in most females. Sexual experience and learning have been suggested as important factors to explain the variability of the female responses. In experiment 1, we compared the behavioral and endocrine responses of four groups of anestrous females that differed in age (young or adult) and previous exposure to males [naive (no exposure) or experienced (courtship behavior for young and numerous mating for adults)]. Age did not seem to affect the LH response to males or their odor. In contrast, sexual experience was a critical factor: the proportion of females exhibiting an LH response to male odor was significantly higher in experienced than in naive ewes. Sexual experience affected the response to male odor, but did not have an effect on responses to the male himself. A second experiment investigated whether the LH response to male odor could result from an associative learning process. Accordingly, we tested the effectiveness of a conditioned stimulus (lavender odor) previously associated with the male, in inducing the endocrine response. The results indicate that the odor of lavender activated LH secretion only in ewes that have been previously exposed to scented males. This demonstrates that ewes are able to learn the association between a neutral odor and their sexual partner. PMID- 15337682 TI - Usefulness of the odor stick identification test for Japanese patients with olfactory dysfunction. AB - The odor stick identification test (OSIT) is a new test of olfactory function recently developed for Japanese people. The purpose of the present study was to evaluate this test in relation to T&T olfactometry and the cross-cultural smell identification test (CC-SIT) by applying to 110 Japanese patients with olfactory disturbance. The averaged recognition thresholds for five odorants in T&T olfactometry, the number of correct answers in the CC-SIT and the rates of identification of 13 odorants in the OSIT were compared. The visual analogue scale (VAS) was also used to evaluate symptoms. The rate of identification of OSIT showed high and significant correlation coefficients with the averaged recognition thresholds of T&T olfactometry (-0.766, P < 0.001), with the number of correct answers in CC-SIT (0.754, P < 0.001) and with the VAS score (0.591, P < 0.001). In addition, on the identification performance measured by OSIT, we found significant differences between all pairs of four degrees of olfactory dysfunction except for one pair. Thus, we conclude that OSIT is useful for evaluating olfactory dysfunction in Japanese people. PMID- 15337683 TI - Diverse bitter stimuli elicit highly similar patterns of Fos-like immunoreactivity in the nucleus of the solitary tract. AB - Previous studies have demonstrated that oral stimulation with quinine elicits Fos like immunoreactivity in the first-order gustatory nucleus, the NST, with a different topographic distribution than sucrose or citric acid. However, it is unknown whether the quinine pattern is unique to this alkaloid or common across bitter stimuli with different chemical structures. Indeed, recent physiological experiments suggest that taste receptor cells and primary afferent neurons may exhibit selectivity for various bitter tastants. The present investigation compared the distribution of FLI in NST following stimulation with three bitter chemicals: QHCl, denatonium and propylthiouracil, stimuli that evoked Ca(2+) currents in almost entirely different sets of receptor cells. The results demonstrate that the quinine pattern is not idiosyncratic but instead generalizes to the other two tastants. Although it remains possible that intermingled but different NST neurons are activated by these stimuli, these data suggest that a specialized region in the NST is preferentially involved in processing a common aspect of bitter tastants. In contrast to citric acid, quinine, denatonium and propylthiouracil all elicited vigorous oromotor rejection responses, consistent with our earlier hypothesis that the medial third of the NST may be an afferent trigger zone for oromotor rejection. PMID- 15337684 TI - Identification of ligands for two human bitter T2R receptors. AB - Earlier, a family of G protein-coupled receptors, termed T2Rs, was identified in the rodent and human genomes through data mining. It was suggested that these receptors mediate bitter taste perception. Analysis of the human genome revealed that the hT2R family is composed of 25 members. However, bitter ligands have been identified for only three human receptors so far. Here we report identification of two novel ligand-receptor pairs. hT2R61 is activated by 6-nitrosaccharin, a bitter derivative of saccharin. hT2R44 is activated by denatonium and 6 nitrosaccharin. Activation profiles for these receptors correlate with psychophysical data determined for the bitter compounds in human studies. Functional analysis of hT2R chimeras allowed us to identify residues in extracellular loops critical for receptor activation by ligands. The discovery of two novel bitter ligand-receptor pairs provides additional support for the hypothesis that hT2Rs mediate a bitter taste response in humans. PMID- 15337685 TI - Predicted 3-D structures for mouse I7 and rat I7 olfactory receptors and comparison of predicted odor recognition profiles with experiment. AB - The first step in the perception of an odor is the activation of one or more olfactory receptors (ORs) following binding of the odorant molecule to the OR. In order to initiate the process of determining how the molecular level receptor odorant interactions are related to odor perception, we used the MembStruk computational method to predict the three-dimensional (3-D) structure of the I7 OR for both mouse and rat. We then used the HierDock ligand docking computational method to predict the binding site and binding energy for the library of 56 odorants to these receptors for which experiment response data are now available. We find that the predicted 3-D structures of the mouse and rat I7 OR lead to predictions of odorant binding that are in good agreement with the experimental results, thus validating the accuracy of both the 3-D structure and the predicted binding site. In particular we predict that heptanal and octanal both bind strongly to both mouse and rat I7 ORs, which conflicts with the older literature but agrees with recent experiments. To provide the basis of additional validations of our 3-D structures, we also report the odorant binding site for a new odorant (8-hydroxy-octanal) with a novel functionality designed to bind strongly to mouse I7. Such validated computational methods should be very useful in predicting the structure and function of many other ORs. PMID- 15337686 TI - 'Thermal taste' predicts higher responsiveness to chemical taste and flavor. AB - Individual differences in taste perception have been explained in part by variations in peripheral innervation associated with the genetic ability to taste the bitter substances PTC and PROP. In the present study we report evidence of another source of individual differences that is independent of taste stimulus, taste quality, or gustatory nerve. Individuals who perceived taste from thermal stimulation alone (thermal taste) gave significantly higher taste ratings to chemical stimuli--often by a factor of >2:1--than did individuals who perceived no taste from thermal stimulation. This was true for all taste stimuli tested (sucrose, saccharin, sodium chloride, citric acid, quinine sulfate, MSG and PROP), for all three gustatory areas of the mouth (anterior tongue, posterior tongue and soft palate) and for whole-mouth stimulation. Moreover, the same individuals reported stronger sensations from the olfactory stimulus vanillin, particularly when it was sensed retronasally. The generality of the thermal taster advantage and its extension to an olfactory stimulus suggests that it arises from individual differences in CNS processes that are involved in perception of both taste and flavor. PMID- 15337687 TI - Gender differences in chemosensory perception and event-related potentials. AB - The present study investigated chemosensory gender differences by means of ratings of total nasal chemosensory intensity, unpleasantness and sensory irritation and simultaneous recordings of chemosensory event-related potentials (CSERPs) for three concentrations of the olfactory/trigeminal stimulus pyridine in 19 women and 17 men, all young adults. Results show that, compared to men, women gave higher intensity and unpleasantness ratings, in particular for the highest stimulus concentration. The gender differences in perceived intensity are reflected in the signal-to-noise ratio of the individual CSERP averages, revealing more identifiable early components (P1, N1) in women than in men. The late positive component, labeled P2/P3, displayed larger amplitudes at all electrode sites and shorter latencies at Cz, in women compared to men. The effects of increased pyridine concentration on perception (larger in women) and CSERPs (similar across gender) imply that the two measures involve partially different neural processing. CSERP component identifiability is proposed here as a general means of assessing signal-to-noise ratio of the CSERPs. PMID- 15337688 TI - Stimulus processing of glycine is dissociable from that of sucrose and glucose based on behaviorally measured taste signal detection in Sac 'taster' and 'non taster' mice. AB - Mouse strains have been divided into 'tasters' and 'non-tasters' based on their relatively high and low preference, respectively, for low concentrations of sucrose and saccharin. These phenotypic differences appear to be due to a polymorphism in the gene at the Sac locus encoding for the T1R3 taste receptor selectively affecting the functionality of the T1R2+3 heterodimer. To psychophysically examine whether these phenotypes are due to sensory sensitivity as opposed to hedonic responsiveness, we measured taste signal detection of sucrose, glucose, and glycine by Sac taster (C57BL/6J and SWR/J) and non-taster (129P3/J and DBA/2J) strains in an operant conditioning paradigm using a gustometer. The taster mice had lower detection thresholds for sucrose and glucose compared with the non-taster mice. The detection thresholds corresponded well with reported responsiveness to low concentrations of these sugars in two bottle intake tests suggesting that the Sac taster phenotype has a sensory basis and is not simply a matter of strain differences in the hedonic evaluation of weak intensities of the stimuli. Taster status did not entirely account for the strain differences in detection thresholds for glycine, a 'sweet' tasting amino acid. Collapsed across strains, detection thresholds for sucrose and glucose were highly correlated with each other (r = 0.81), but only modestly correlated with those for glycine (r < or = 0.43). This suggests that stimulus processing of glycine in the perithreshold intensity domain can be dissociated from that of sucrose and glucose. The mechanism underlying this difference may be related to the ability of glycine to bind with the T1R1+3 heterodimer. PMID- 15337689 TI - Regulated on activation, normal T cell expressed and secreted (RANTES) antagonist (Met-RANTES) controls the early phase of Trypanosoma cruzi-elicited myocarditis. AB - BACKGROUND: Comprehension of the pathogenesis of Trypanosoma cruzi-elicited myocarditis is crucial to delineate strategies aimed at ameliorating the inflammation associated with heart dysfunction. The augmented expression of CC chemokines, especially CCL5/RANTES and CCL3/MIP-1alpha, in the hearts of infected mice suggests a role for CC chemokines and their receptors in the pathogenesis of T cruzi-elicited myocarditis. METHODS AND RESULTS: We report that during the early phase of infection in C3H/HeJ mice infected with 100 blood trypomastigotes of T cruzi, most of the inflammatory cells invading the heart tissue were CD8+ cells and expressed CCR5, a CCL5/RANTES, and CCL3/MIP1-alpha receptor. Furthermore, peripheral blood CD8+ T lymphocytes displayed increased expression of CCR5. These findings led us to use Met-RANTES, a selective CCR1 and CCR5 antagonist, to modulate the acute T cruzi-elicited myocarditis. Met-RANTES treatment did not interfere with parasitism but significantly decreased the numbers of CD4+ and CD8+ T cells, CCR5+, and interleukin-4+ cells invading the heart, paralleling the diminished deposition of fibronectin. Moreover, Met-RANTES treatment resulted in increased survival of infected animals, compared with saline treatment. CONCLUSIONS: These results indicate that the massive influx of CCR5+ cells into cardiac tissue is not crucial for cell-mediated anti-T cruzi immunity but appears to be critical for pathogenesis of T cruzi-elicited myocarditis. Thus, CC chemokine receptors might become an attractive therapeutic target for further evaluation during T cruzi infection. PMID- 15337690 TI - Explaining the increase in coronary heart disease mortality in Beijing between 1984 and 1999. AB - BACKGROUND: Coronary heart disease (CHD) mortality is rising in many developing countries. We examined how much of the increase in CHD mortality in Beijing, China, between 1984 and 1999 could be attributed to changes in major cardiovascular risk factors and assessed the impact of medical and surgical treatments. METHODS AND RESULTS: A validated, cell-based mortality model synthesized data on (1) patient numbers, (2) uptake of specific medical and surgical treatments, (3) treatment effectiveness, and (4) population trends in major cardiovascular risk factors (smoking, total cholesterol, blood pressure, obesity, and diabetes). Main data sources were the WHO MONICA and Sino-MONICA studies, the Chinese Multi-provincial Cohort Study, routine hospital statistics, and published meta-analyses. Age-adjusted CHD mortality rates increased by approximately 50% in men and 27% in women (1608 more deaths in 1999 than expected by application of 1984 rates). Most of this increase ( approximately 77%, or 1397 additional deaths) was attributable to substantial rises in total cholesterol levels (more than 1 mmol/L), plus increases in diabetes and obesity. Blood pressure decreased slightly, whereas smoking prevalence increased in men but decreased substantially in women. In 1999, medical and surgical treatments in patients together prevented or postponed approximately 642 deaths, mainly from initial treatments for acute myocardial infarction ( approximately 41%), hypertension (24%), angina (15%), secondary prevention (11%), and heart failure (10%). Multiway sensitivity analyses did not greatly influence the results. CONCLUSIONS: Much of the dramatic CHD mortality increases in Beijing can be explained by rises in total cholesterol, reflecting an increasingly "Western" diet. Without cardiological treatments, increases would have been even greater. PMID- 15337691 TI - Long-term outcome in elderly patients with chronic angina managed invasively versus by optimized medical therapy: four-year follow-up of the randomized Trial of Invasive versus Medical therapy in Elderly patients (TIME). AB - BACKGROUND: There are no prospective trial data on long-term outcomes in 80-year old patients with chronic angina with regard to antiischemic therapy. METHODS AND RESULTS: To assess long-term survival and quality of life (QoL) in patients from the Trial of Invasive versus Medical Therapy in the Elderly (TIME), all 276 1 year survivors (of a total 301 patients) were contacted after a median of 3.1 years (range, 1.1 to 5.9 years). At baseline, patients were 80+/-4 years old, 42% were women, and they were designated as being in angina class 3.2+/-0.7, despite their taking 2.5+/-0.7 antiischemic drugs. Patients were randomized to an invasive (n=153) or an optimized medical (n=148) strategy. Survival of invasive strategy versus medical-strategy patients was 91.5% versus 95.9% after 6 months, 89.5% versus 93.9% after 1 year, and 70.6% versus 73.0% after 4.1 years (P=NS). Mortality was independently increased in patients >or=80 years of age, with prior heart failure, ejection fraction or=2 comorbidities, and without revascularization within the first year. Revascularization within the first year improved survival in invasive-strategy (P=0.07) and medical-strategy (P<0.001) patients. The early benefit of both treatments in angina relief and QoL was maintained long term, but freedom from major events remained higher in invasive strategy versus medical-strategy patients (39% versus 20%, P<0.0001). CONCLUSIONS: Long-term survival was similar for patients assigned to invasive and medical treatment. The benefits of both treatments in angina relief and improvement in QoL were maintained, but nonfatal events occurred more frequently in patients assigned to medical treatment. Irrespective of whether patients were catheterized initially or only after drug therapy failure, their survival rates were better if they were revascularized within the first year. PMID- 15337692 TI - Simvastatin induces heme oxygenase-1: a novel mechanism of vessel protection. AB - BACKGROUND: Evidence from experimental and clinical studies indicates that statins can protect the vessel wall through cholesterol-independent mechanisms. The "pleiotropic" effects include the prevention of inflammation and proliferation of vascular cells. Here, we studied whether heme oxygenase-1 (HO 1), an important cytoprotective molecule, is induced by simvastatin and the role of HO-1 in the pleiotropic effects of simvastatin. METHODS AND RESULTS: Human and rat aortic smooth muscle cells treated with simvastatin showed an elevated level of HO-1 for up to 24 hours. The induction of HO-1 by simvastatin was not found in cultured endothelial cells and macrophages. Injecting C57BL/6J mice intraperitoneally with simvastatin increased the level of HO-1 in vascular SMCs (VSMCs) in the tunica media. Treating VSMCs with zinc protoporphyrin, an HO-1 inhibitor, or HO-1 small interfering RNA (siRNA) blocked the antiinflammatory effect of simvastatin, including the inhibition of nuclear factor-kappaB activation and nitric oxide production. Blockade of HO-1 also abolished the simvastatin-induced p21(Waf1) and the associated antiproliferative effect. Simvastatin activated p38 and Akt in VSMCs, and the respective inhibitors of p38 and phosphoinositide 3-kinase (PI3K) greatly reduced the level of simvastatin induced HO-1, which suggests the involvement of p38 and the PI3K-Akt pathway in HO-1 induction. CONCLUSIONS: Simvastatin activates HO-1 in VSMCs in vitro and in vivo. The antiinflammatory and antiproliferative effects of simvastatin occur largely through the induced HO-1. PMID- 15337693 TI - Beyond restenosis: five-year clinical outcomes from second-generation coronary stent trials. AB - BACKGROUND: In the first year after coronary stent implantation, clinical failures are driven mainly by procedural complications and restenosis, but the subsequent relative contributions of restenosis and disease progression to late failures are less clear. METHODS AND RESULTS: We observed 1228 patients for 5 years after the implantation of stents as part of pivotal second-generation coronary stent trials. Clinical events of death, myocardial infarction, repeat revascularization, and repeat hospitalization for acute coronary syndrome or congestive heart failure were attributed to the index stented (target) lesion or other distinct sites (either in the target or other coronary vessels) and further classified as procedural, restenosis, or nonrestenosis. During the first year the hazard rate was 18.3% for target-lesion events and 12.4% for events unrelated to the target lesion. After the first year the average annual hazard rate was 1.7% for target-lesion events and 6.3% for nontarget-lesion events. By the fifth year, restenosis events occurred in 20.3% of patients, whereas 30-day procedural complications or later nonrestenosis events occurred in 37.9%, including 11.4% who also experienced a restenosis event, for a combined cumulative event rate of 46.4%. Diabetes mellitus and multivessel disease were independently associated with increased risk for both restenosis and nonrestenosis events. CONCLUSIONS: In a low-risk clinical trial population, the clinical outcome beyond 1 year after stenting is determined by a high rate of events related to disease progression in segments other than the stented lesion, which itself remains relatively stable. PMID- 15337694 TI - Plasma levels of cholesteryl ester transfer protein and the risk of future coronary artery disease in apparently healthy men and women: the prospective EPIC (European Prospective Investigation into Cancer and nutrition)-Norfolk population study. AB - BACKGROUND: Low plasma levels of cholesteryl ester transfer protein (CETP) are associated with elevated levels of HDL cholesterol (HDL-C), but it remains unclear whether this translates into a concomitant reduction in the risk of coronary artery disease (CAD). Evidence exists that the effect of CETP depends on metabolic context, in particular on triglyceride levels. METHODS AND RESULTS: A nested case-control study was performed in the prospective EPIC-Norfolk cohort study. Cases were apparently healthy men and women aged 45 to 79 years who developed fatal or nonfatal CAD during follow-up. Control subjects were matched by age, sex, and enrollment time. CETP levels were not significantly different between cases and controls (4.0+/-2.2 versus 3.8+/-2.1 mg/L, P=0.07). CETP levels were significantly related to plasma levels of total cholesterol, LDL cholesterol, and HDL-C. The risk of CAD increased with increasing CETP quintiles (P for linearity=0.02), such that subjects in the highest quintile had an adjusted OR of 1.43 (95% CI 1.03 to 1.99, P=0.03) versus those in the lowest. Among individuals with triglyceride levels below the median (1.7 mmol/L), no relationship between CETP levels and CAD risk was observed (P for linearity=0.5), but this relationship was strong among those with high triglyceride levels (P for linearity=0.02), such that those in the highest CETP quintile had an OR of 1.87 (95% CI 1.06 to 3.30, P=0.02). CONCLUSIONS: Elevated CETP levels are associated with an increasing risk of future CAD in apparently healthy individuals, but only in those with high triglyceride levels. PMID- 15337695 TI - Nesiritide does not improve renal function in patients with chronic heart failure and worsening serum creatinine. AB - BACKGROUND: Nesiritide (synthetic human brain natriuretic peptide) is approved for the treatment of symptomatic heart failure. However, studies of brain natriuretic peptide in patients with heart failure have come to conflicting conclusions about effects on glomerular filtration rate (GFR), effective renal plasma flow, natriuresis, and diuresis. METHODS AND RESULTS: To identify a population at high risk of renal dysfunction with conventional treatment, we selected patients with a creatinine level increased from baseline (within 6 months). We examined the effects of nesiritide on GFR (measured by iothalamate clearance), renal plasma flow (measured by para-amino hippurate clearance), urinary sodium excretion, and urine output in a double-blind, placebo-controlled, crossover study. Patients received nesiritide (2 microg/kg IV bolus followed by an infusion of 0.01 microg/kg per minute) or placebo for 24 hours on consecutive days. Nesiritide and placebo data were compared by repeated-measures analysis and Student t test. We studied 15 patients with a recent mean baseline creatinine of 1.5+/-0.4 mg/dL and serum creatinine of 1.8+/-0.8 mg/dL on admission to the study. There were no differences in GFR, effective renal plasma flow, urine output, or sodium excretion for any time interval or for the entire 24-hour period between the nesiritide and placebo study days. For 24 hours, urine output was 113+/-51 mL/h with placebo and 110+/-56 mL/h with nesiritide. GFR during placebo was 40.9+/-25.9 mL/min and with nesiritide was 40.9+/-25.8. CONCLUSIONS: Nesiritide did not improve renal function in patients with decompensated heart failure, mild chronic renal insufficiency, and renal function that had worsened compared with baseline. The lack of effect may be related to renal insufficiency, hemodynamic alterations, sodium balance, severity of heart failure, or drug dose. Understanding the importance of these issues will permit effective and appropriate use of nesiritide. PMID- 15337696 TI - Norepinephrine transporter-deficient mice exhibit excessive tachycardia and elevated blood pressure with wakefulness and activity. AB - BACKGROUND: Norepinephrine (NE) is a primary neurotransmitter of central autonomic regulation and sympathetic nerve conduction, and the norepinephrine transporter (NET) is crucial in limiting catecholaminergic signaling. NET is sensitive to antidepressants, cocaine, and amphetamine. NET blockade often is associated with cardiovascular side effects, and NET deficiency is linked to tachycardia in familial orthostatic intolerance. METHODS AND RESULTS: We telemetrically monitored NET-deficient (NET(-/-)) mice to determine the cardiovascular effects of reduced NE reuptake. Mean arterial pressure was elevated in resting NET(-/-) mice compared with NET(+/+) controls (103+/-0.6 versus 99+/-0.4 mm Hg; P<0.01), and corresponding pressures increased to 122+/ 0.3 and 116+/-0.3 mm Hg (P<0.0001) with activity. Heart rate was also greater in resting NET(-/-) mice (565+/-5 versus 551+/-3 bpm; P<0.05), and genotypic differences were highly significant during the active phase (640+/-5 versus 607+/ 3 bpm; P<0.0001). Conversely, the respiratory rate of resting NET(-/-) mice was dramatically reduced, whereas increases after the day/night shift surpassed those of controls. Plasma catecholamines in NET(-/-) and NET(+/+) mice were as follows: NE, 69+/-8 and 32+/-7; dihydroxyphenylglycol, 2+0.4 and 17+/-3; epinephrine, 15+/ 3 and 4+/-0.6; and dopamine, 13+/-4 and 4+/-1 pmol/mL. Catechols in urine, brain, and heart also were determined. CONCLUSIONS: Resting mean arterial pressure and heart rate are maintained at nearly normal levels in NET-deficient mice, most likely as a result of increased central sympathoinhibition. However, sympathetic activation with wakefulness and activity apparently overwhelms central modulation, amplifying peripheral catecholaminergic signaling, particularly in the heart. PMID- 15337697 TI - Thioredoxin-1 ameliorates myosin-induced autoimmune myocarditis by suppressing chemokine expressions and leukocyte chemotaxis in mice. AB - BACKGROUND: Cardiac myosin-induced myocarditis is an experimental autoimmune myocarditis (EAM) model used to investigate autoimmunological mechanisms in inflammatory heart diseases and resembles fulminant myocarditis in humans. We investigated the therapeutic role of thioredoxin-1 (TRX-1), a redox-regulatory protein with antioxidant and antiinflammatory effects, in murine EAM. METHODS AND RESULTS: EAM was generated in 5-week-old male BALB/c mice by immunization with porcine cardiac myosin at days 0 and 7. Recombinant human TRX-1 (rhTRX-1), C32S/C35S mutant rhTRX-1, or saline was administered intraperitoneally every second day from day 0 to 20. In addition, rabbit anti-mouse TRX-1 serum or normal rabbit serum was administered intraperitoneally on days -1, 2, and 6. Animals were euthanized on day 21. Histological analysis of the heart showed that TRX-1 significantly reduced the severity of EAM, whereas mutant TRX-1 failed to have such an effect, and anti-TRX-1 antibody enhanced the disease markedly. Immunohistochemical analysis showed that TRX-1 significantly suppressed cardiac macrophage inflammatory protein (MIP)-1alpha, MIP-2, and 8-hydroxydeoxyguanosine expression and macrophage infiltration into the heart in EAM. Although serum levels of MIP-1alpha were not suppressed by TRX-1 until day 21, both an in vitro chemotaxis chamber assay and an in vivo air pouch model showed that TRX-1 significantly suppressed MIP-1alpha- or MIP-2-induced leukocyte chemotaxis. However, real-time reverse transcription-polymerase chain reaction showed that TRX-1 failed to decrease chemokine receptor expression increased in the bone marrow cells of EAM mice. CONCLUSIONS: TRX-1 attenuates EAM by suppressing chemokine expressions and leukocyte chemotaxis in mice. PMID- 15337698 TI - Natriuretic peptide receptor-C regulates coronary blood flow and prevents myocardial ischemia/reperfusion injury: novel cardioprotective role for endothelium-derived C-type natriuretic peptide. AB - BACKGROUND: Ischemia/reperfusion (I/R) injury complicates myocardial infarction and stroke by exacerbating tissue damage and increasing risk of mortality. We have recently identified C-type natriuretic peptide (CNP) as an endothelium derived hyperpolarizing factor in the mesenteric resistance vasculature and described a novel signaling pathway involving activation of natriuretic peptide receptor C (NPR-C), which plays a pivotal role in the regulation of local blood flow. We tested the hypothesis that CNP/NPR-C signaling is a novel regulatory pathway governing coronary blood flow and protecting against I/R injury. METHODS AND RESULTS: CNP and (Cys18)-atrial natriuretic factor (4-23) amide (cANF(4-23)) elicited dose-dependent decreases in coronary perfusion pressure (CPP) that were blocked by Ba(2+) and ouabain in the isolated Langendorff rat heart. The endothelium-dependent vasodilator acetylcholine elicited the release of CNP from the coronary endothelium. CNP and cANF(4-23) reduced infarct size after 25 minutes of global ischemia and 120 minutes of reperfusion, maintaining CPP and left ventricular pressure at preischemic values. The vasorelaxant and protective activity of CNP and cANF(4-23) were enhanced in the absence of endothelium derived nitric oxide. CONCLUSIONS: Endothelium-derived CNP is involved in the regulation of the coronary circulation, and NPR-C activation underlies the vasorelaxant activity of this peptide. Moreover, this newly defined pathway represents a protective mechanism against I/R injury and a novel target for therapeutic intervention in ischemic cardiovascular disorders. PMID- 15337699 TI - Synergistic effect of bone marrow mobilization and vascular endothelial growth factor-2 gene therapy in myocardial ischemia. AB - BACKGROUND: We performed a series of investigations to test the hypothesis that combining angiogenic gene therapy and cytokine (CK)-induced endothelial progenitor cell mobilization would be superior to either strategy alone for treatment of chronic myocardial ischemia. METHODS AND RESULTS: A swine model of chronic myocardial ischemia and a murine model of acute myocardial infarction were used in this study. In both models, animals were randomly assigned to 1 of 4 treatment groups: Combo group, intramyocardial vascular endothelial growth factor (VEGF)-2 gene transfer plus subcutaneous injection of CKs; VEGF-2, VEGF-2 gene transfer plus saline subcutaneously injected; CK, empty vector transfer plus CKs; and control, empty vector plus subcutaneous saline. Acute myocardial infarction was also induced in wild-type mice 4 weeks after bone marrow transplantation from enhanced green fluorescent protein transgenic mice to permit observation of bone marrow-derived cells in the myocardium after acute myocardial infarction. In chronic myocardial ischemia, combination therapy resulted in superior improvement in all indexes of perfusion and function compared with all other treatment groups. In the bone marrow transplant mice, double immunofluorescent staining revealed that the combination of CK-induced mobilization and local VEGF-2 gene transfer resulted in a significant increase in the number of bone marrow-derived cells incorporating into the neovasculature, indicating that recruitment and/or retention of bone marrow-derived progenitors was enhanced by mobilization and that local VEGF-2 gene transfer can provide signals for recruitment or incorporation of circulating progenitor cells. CONCLUSIONS: Mobilization of endothelial progenitor cells with cytokines potentiates VEGF-2 gene therapy for myocardial ischemia and enhances bone marrow cell incorporation into ischemic myocardium. PMID- 15337700 TI - Effect of baseline or changes in adrenergic activity on clinical outcomes in the beta-blocker evaluation of survival trial. AB - BACKGROUND: Adrenergic activation is thought to be an important determinant of outcome in subjects with chronic heart failure (CHF), but baseline or serial changes in adrenergic activity have not been previously investigated in a large patient sample treated with a powerful antiadrenergic agent. METHODS AND RESULTS: Systemic venous norepinephrine was measured at baseline, 3 months, and 12 months in the beta-Blocker Evaluation of Survival Trial (BEST), which compared placebo treatment with the beta-blocker/sympatholytic agent bucindolol. Baseline norepinephrine level was associated with a progressive increase in rates of death or death plus CHF hospitalization that was independent of treatment group. On multivariate analysis, baseline norepinephrine was also a highly significant (P<0.001) independent predictor of death. In contrast, the relation of the change in norepinephrine at 3 months to subsequent clinical outcomes was complex and treatment group-dependent. In the placebo-treated group but not in the bucindolol treated group, marked norepinephrine increase at 3 months was associated with increased subsequent risks of death or death plus CHF hospitalization. In the bucindolol-treated group but not in the placebo-treated group, the 1st quartile of marked norepinephrine reduction was associated with an increased mortality risk. A likelihood-based method indicated that 18% of the bucindolol group but only 1% of the placebo group were at an increased risk for death related to marked reduction in norepinephrine at 3 months. CONCLUSIONS: In BEST, a subset of patients treated with bucindolol had an increased risk of death as the result of sympatholysis, which compromised the efficacy of this third-generation beta blocker. PMID- 15337701 TI - Homozygous deficiency of heparin cofactor II: relevance of P17 glutamate residue in serpins, relationship with conformational diseases, and role in thrombosis. AB - BACKGROUND: Heparin cofactor II (HCII) is a hepatic serpin with significant antithrombin activity that has been implicated in coagulation, inflammation, atherosclerosis, and wound repair. Recent data obtained in mice lacking HCII suggest that this serpin might inhibit thrombosis in the arterial circulation. However, the clinical relevance and molecular mechanisms associated with deficiency of HCII in humans are unclear. METHODS AND RESULTS: We studied the first family with homozygous HCII deficiency, identifying a Glu428Lys mutation affecting a conserved glutamate at the hinge (P17) of the reactive loop. No carrier reported arterial thrombosis, and only 1 homozygous HCII-deficient patient developed severe deep venous thrombosis, but she also had a de novo Glu100Stop nonsense truncation in the antithrombin gene. CONCLUSIONS: Our results confirm the key structural role of the P17 glutamate in serpins. The same mutation causes conformational instability and polymerization in 3 serpins: Drosophila necrotic, human alpha1-antitrypsin, and human HCII, which explains their plasma deficiency. In the family under study here, however, plasma HCII deficiency was not associated with a significant clinical phenotype. PMID- 15337702 TI - Subxiphoid surgical approach for epicardial catheter-based mapping and ablation in patients with prior cardiac surgery or difficult pericardial access. AB - BACKGROUND: Percutaneous epicardial mapping and ablation are successful in some patients with ventricular epicardial reentry circuits but may be impossible when pericardial adhesions are present, such as from prior cardiac surgery. The purpose of this study was to evaluate the feasibility of direct surgical exposure of the pericardial space to allow catheter epicardial mapping and ablation in the electrophysiology laboratory when percutaneous access is not feasible. METHODS AND RESULTS: In 6 patients with prior cardiac surgery or failed percutaneous pericardial access, a subxiphoid pericardial window was attempted. In all 6 patients, manual lysis of adhesions exposed the epicardial surface of the heart through a small subxiphoid incision and allowed placement of an 8F sheath into the pericardial space under direct vision. Access to the diaphragmatic surface of the heart with ablation catheters was achieved in all patients, and catheter manipulation to the lateral and anterior walls was possible in 4 patients. Three dimensional electroanatomic voltage maps revealed low-amplitude regions in the inferior or posterior left ventricular epicardium. A total of 16 ventricular tachycardias were induced, and 14 were abolished by radiofrequency ablation. Ablation was limited by intrapericardial defibrillator patches adherent to the likely target region in 2 patients. All patients had chest pain consistent with pericarditis early after the procedure that resolved within a few days. There were no other complications. CONCLUSIONS: A direct surgical subxiphoid epicardial approach in the electrophysiology laboratory is feasible for patients with difficult pericardial access who require ablation of epicardial arrhythmia foci. PMID- 15337703 TI - Increased sympathetic nerve activity in pulmonary artery hypertension. AB - BACKGROUND: This study tested the hypothesis that sympathetic nerve activity is increased in pulmonary artery hypertension (PAH), a rare disease of poor prognosis and incompletely understood pathophysiology. We subsequently explored whether chemoreflex activation contributes to sympathoexcitation in PAH. METHODS AND RESULTS: We measured muscle sympathetic nerve activity (MSNA) by microneurography, heart rate (HR), and arterial oxygen saturation (Sao(2)) in 17 patients with PAH and 12 control subjects. The patients also underwent cardiac echography, right heart catheterization, and a 6-minute walk test with dyspnea scoring. Circulating catecholamines were determined in 8 of the patients. Chemoreflex deactivation by 100% O(2) was assessed in 14 patients with the use of a randomized, double-blind, placebo-controlled, crossover study design. Compared with the controls, the PAH patients had increased MSNA (67+/-4 versus 40+/-3 bursts per minute; P<0.0001) and HR (82+/-4 versus 68+/-3 bpm; P=0.02). MSNA in the PAH patients was correlated with HR (r=0.64, P=0.006), Sao(2) (r=-0.53, P=0.03), the presence of pericardial effusion (r=0.51, P=0.046), and NYHA class (r=0.52, P=0.033). The PAH patients treated with prostacyclin derivatives had higher MSNA (P=0.009), lower Sao(2) (P=0.01), faster HR (P=0.003), and worse NYHA class (P=0.04). Plasma catecholamines were normal. Peripheral chemoreflex deactivation with hyperoxia increased Sao(2) (91.7+/-1% to 98.4+/-0.2%; P<0.0001) and decreased MSNA (67+/-5 to 60+/-4 bursts per minute; P=0.0015), thereby correcting approximately one fourth of the difference between PAH patients and controls. CONCLUSIONS: We report for the first time direct evidence of increased sympathetic nerve traffic in advanced PAH. Sympathetic hyperactivity in PAH is partially chemoreflex mediated and may be related to disease severity. PMID- 15337704 TI - Statins but not angiotensin-converting enzyme inhibitors delay progression of aortic stenosis. AB - BACKGROUND: Recently, statins and angiotensin-converting enzyme inhibitors (ACEIs) have been shown to slow aortic valve calcium accumulation. Although several studies also suggest that statins may reduce the hemodynamic progression of aortic stenosis (AS), no data are available for ACEIs or the combination of both. METHODS AND RESULTS: A total of 211 consecutive patients (aged 70+/-10 years, 104 females) with native AS, defined by a peak velocity >2.5 m/s (valve area 0.84+/-0.23 cm(2), mean gradient 42+/-19 mm Hg), with normal left ventricular function and no other significant valvular lesion who were examined between 2000 and 2002 and who had 2 echocardiograms separated by at least 6 months were included. Of these, 102 patients were treated with ACEIs, 50 patients received statins, and 32 patients received both. Hemodynamic progression of AS was assessed and related to medical treatment. Annualized increase in peak aortic jet velocity for the entire study group was 0.32+/-0.44 m x s(-1) x y(-1). Progression was significantly lower in patients treated with statins (0.10+/-0.41 m x s(-1) x y(-1)) than in those who were not (0.39+/-0.42 m x s(-1) x y(-1); P<0.0001). This effect was observed both in mild-to-moderate and severe AS. ACEI use, however, did not significantly affect hemodynamic progression (P=0.29). Furthermore, ACEIs had no additional effect on AS progression when given in combination with statins (0.11+/-0.42 versus 0.08+/-0.43 m x s(-1) x y(-1) for combination versus statin only; P=0.81). Cholesterol levels did not correlate with hemodynamic progression either in the group receiving statins or in the group that did not. CONCLUSIONS: ACEIs do not appear to slow AS progression. However, statins significantly reduce the hemodynamic progression of both mild-to moderate and severe AS, an effect that may not be related to cholesterol lowering. PMID- 15337705 TI - Modulating angiogenesis: the yin and the yang in ginseng. AB - BACKGROUND: Ginseng is a commonly used nutraceutical. Intriguingly, existing literature reports both wound-healing and antitumor effects of ginseng extract through opposing activities on the vascular system. To elucidate this perplexity, we merged a chemical fingerprinting approach with a deconstructional study of the effects of pure molecules from ginseng extract on angiogenesis. METHODS AND RESULTS: A mass spectrometric compositional analysis of American, Chinese and Korean, and Sanqi ginseng revealed distinct "sterol ginsenoside" fingerprints, especially in the ratio between a triol, Rg1, and a diol, Rb1, the 2 most prevalent constituents. Using a Matrigel implant model and reconstituting the extracts using distinct ratios of the 2 ginsenosides, we demonstrate that the dominance of Rg1 leads to angiogenesis, whereas Rb1 exerts an opposing effect. Rg1 also promoted functional neovascularization into a polymer scaffold in vivo and the proliferation of, chemoinvasion of, and tubulogenesis by endothelial cells in vitro, an effect mediated through the expression of nitric oxide synthase and the phosphatidylinositol-3 kinase-->Akt pathway. In contrast, Rb1 inhibited the earliest step in angiogenesis, the chemoinvasion of endothelial cells. CONCLUSIONS: The present study explains, for the first time, the ambiguity about the effects of ginseng in vascular pathophysiology based on the existence of opposing active principles in the extract. We also unraveled a speciogeographic variation impinging on the compositional fingerprint that may modulate the final phenotype. This emphasizes the need for regulations standardizing herbal therapy, currently under the Dietary Supplement and Health Education Act. Furthermore, we propose that Rg1 could be a prototype for a novel group of nonpeptide molecules that can induce therapeutic angiogenesis, such as in wound healing. PMID- 15337706 TI - Nosocomial infections: what needs to be done? PMID- 15337707 TI - Lead poisoning in children. PMID- 15337708 TI - Lead poisoning in children. PMID- 15337710 TI - Possible causes of cognitive decline. PMID- 15337713 TI - Credentialing body needed for foreign-trained doctors. PMID- 15337712 TI - Genetics and insurance. PMID- 15337714 TI - C. difficile hits Sherbrooke, Que., hospital: 100 deaths. PMID- 15337715 TI - Eleven million adults co-infected with AIDS, TB. PMID- 15337717 TI - Cross-border malpractice coverage cancelled. PMID- 15337719 TI - Whistleblower protection needed in Canada: advocates. PMID- 15337720 TI - Stem cell therapy for myocardial infarction. PMID- 15337721 TI - Use of telemedicine for an emergency craniotomy in a pediatric trauma. PMID- 15337722 TI - Bacterial vaginosis: more questions than answers. PMID- 15337723 TI - Several Chinese herbal products may contain toxic aristolochic acid. PMID- 15337724 TI - Her tongue tipped us off. PMID- 15337725 TI - Variations in rates of appropriate and inappropriate carotid endarterectomy for stroke prevention in 4 Canadian provinces. AB - BACKGROUND: Carotid endarterectomy (CE), when performed on appropriate patients, reduces the incidence of stroke, yet there are marked variations in rates of this procedure. We sought to determine reasons for the variation in CE rates in 4 Canadian provinces. METHODS: We identified all CEs performed in 4 Canadian provinces between January 2000 and December 2001, inclusive. From chart review and expert assessment, we determined the proportion of these procedures that were appropriate, inappropriate or of uncertain appropriateness, using the RAND/UCLA Appropriateness Method. We sought to determine the variation in rates by province and whether the variation was due to differences in type of hospital, surgical specialty or surgical volume. RESULTS: Overall, 1656 (52.3%) of the 3167 CEs studied were performed for appropriate indications. The proportions of appropriate procedures were 78.2% (176/225) in Saskatchewan, 58.7% (481/819) in Alberta, 49.1% (350/713) in Manitoba and 46.0% (649/1410) in British Columbia (p < 0.001 across provinces). Rates of appropriate procedures per 100 000 population ranged from 44.3 in Manitoba to 16.2 in Saskatchewan (p < 0.001 across provinces). CEs were more likely to be appropriate when performed by a neurosurgeon compared with all other surgeons (74.4% v. 49.4% were appropriate; p < 0.001), when performed by surgeons doing fewer than 31 procedures over 2 years compared with surgeons doing more than 31 (70.1% v. 49.5% were appropriate; p < 0.001) and when performed in hospitals doing fewer than 135 procedures per year compared with hospitals doing more than 135 (63.4% v. 49.1% were appropriate; p < 0.001). Overall, 10.3% of procedures were done for inappropriate reasons. INTERPRETATION: Our findings suggest some overuse (for inappropriate or uncertain indications) but also some underuse (low population rates in some regions). High rates of CE are associated with lower rates of appropriateness for both surgeons and hospitals. That 1 in 10 CEs is done inappropriately suggests the need for preoperative assessment of appropriateness. PMID- 15337726 TI - Association of socioeconomic status and receipt of colorectal cancer investigations: a population-based retrospective cohort study. AB - BACKGROUND: Although the Canadian health care system was designed to ensure equal access, inequities persist. It is not known if inequities exist for receipt of investigations used to screen for colorectal cancer (CRC). We examined the association between socioeconomic status and receipt of colorectal investigation in Ontario. METHODS: People aged 50 to 70 years living in Ontario on Jan. 1, 1997, who did not have a history of CRC, inflammatory bowel disease or colorectal investigation within the previous 5 years were followed until death or Dec. 31, 2001. Receipt of any colorectal investigation between 1997 and 2001 inclusive was determined by means of linked administrative databases. Income was imputed as the mean household income of the person's census enumeration area. Multivariate analysis was performed to evaluate the relationship between the receipt of any colorectal investigation and income. RESULTS: Of the study cohort of 1,664,188 people, 21.2% received a colorectal investigation in 1997-2001. Multivariate analysis demonstrated a significant association between receipt of any colorectal investigation and income (p < 0.001); people in the highest-income quintile had higher odds of receiving any colorectal investigation (adjusted odds ratio [OR] 1.38; 95% confidence interval [CI] 1.36-1.40) and of receiving colonoscopy (adjusted OR 1.50; 95% CI 1.48-1.53). INTERPRETATION: Socioeconomic status is associated with receipt of colorectal investigations in Ontario. Only one-fifth of people in the screening-eligible age group received any colorectal investigation. Further work is needed to determine the reason for this low rate and to explore whether it affects CRC mortality. PMID- 15337727 TI - Clostridium difficile-associated diarrhea in a region of Quebec from 1991 to 2003: a changing pattern of disease severity. AB - BACKGROUND: Recent reports suggest that Clostridium difficile colitis may be evolving into a more severe disease. During the second half of 2002 we noted an increase in the number of patients with severe C. difficile-associated diarrhea (CDAD) in our institution. We describe cases of CDAD at our institution over a 13 year period and investigate changes in illness severity. METHODS: We undertook a retrospective chart review of all cases of CDAD diagnosed at the Centre hospitalier universitaire de Sherbrooke from Jan. 1, 1991, to Dec. 31, 2003. Because the hospital serves a well-defined population of Quebec, we were also able to calculate population-based incidence during this period. We abstracted data on individual patients from patient charts and from hospital and pharmacy computer databases. We defined cases of CDAD as having a positive C. difficile cytotoxicity assay result, or endoscopic or histopathological evidence of pseudomembranous colitis. A case was considered complicated if one or more of the following was observed: megacolon, perforation, colectomy, shock requiring vasopressor therapy, or death within 30 days after diagnosis. RESULTS: A total of 1721 cases of CDAD were diagnosed during the study period. The incidence increased from 35.6 per 100,000 population in 1991 to 156.3 per 100,000 in 2003; among patients aged 65 years or more, it increased from 102.0 to 866.5 per 100,000. The proportion of cases that were complicated increased from 7.1% (12/169) in 1991-1992 to 18.2% (71/390) in 2003 (p < 0.001), and the proportion of patients who died within 30 days after diagnosis increased from 4.7% (8/169) in 1991-1992 to 13.8% (54/390) in 2003 (p < 0.001). A high leukocyte count (20.0 small ha, Cyrillic 10(9)/L or greater) and an elevated creatinine level (200 micromol/L or greater) were strongly associated with adverse outcomes: in 2003, 45 (40.9%) of 110 patients with a high leukocyte count or creatinine level, or both, had complicated CDAD and 28 (25.5%) died within 30 days after diagnosis. After adjustment for age and other confounding factors, patients initially given oral vancomycin therapy had a risk of progression to complicated CDAD that was 79% lower than the risk among patients initially treated with metronidazole (adjusted odds ratio 0.2, 95% confidence interval 0.06-0.8, p = 0.02). INTERPRETATION: An epidemic of CDAD with an increased case-fatality rate has had important consequences on the elderly population of our region. Our observational data suggest that the equivalence of vancomycin and metronidazole in the treatment of CDAD needs to be questioned. PMID- 15337728 TI - The inappropriate use of carotid endarterectomy. PMID- 15337729 TI - Editors' declaration of their own conflicts of interest. PMID- 15337730 TI - Complicated and fatal Strongyloides infection in Canadians: risk factors, diagnosis and management. AB - Strongyloidiasis, which is caused by the nematode Strongyloides stercoralis, is a common and persistent infection, particularly in developing countries. In the setting of compromised cellular immunity, it can result in fulminant dissemination with case-fatality rates of over 70%. The majority of new Canadian immigrants come from countries where Strongyloides is highly endemic; therefore, the burden of Strongyloides may be underappreciated in Canada. Because early diagnosis and therapy can have a marked impact on disease outcome, screening for this infection should be considered mandatory for patients who have a history of travel or residence in a disease-endemic area and risk factors for disseminated disease (e.g., corticosteroid use and human T-lymphotropic virus type I infection). PMID- 15337731 TI - High-dose statins in acute coronary syndromes: not just lipid levels. PMID- 15337732 TI - Early intensive vs a delayed conservative simvastatin strategy in patients with acute coronary syndromes: phase Z of the A to Z trial. AB - CONTEXT: Limited data are available evaluating how the timing and intensity of statin therapy following an acute coronary syndrome (ACS) event affect clinical outcome. OBJECTIVE: To compare early initiation of an intensive statin regimen with delayed initiation of a less intensive regimen in patients with ACS. DESIGN, SETTING, AND PARTICIPANTS: International, randomized, double-blind trial of patients with ACS receiving 40 mg/d of simvastatin for 1 month followed by 80 mg/d thereafter (n = 2265) compared with ACS patients receiving placebo for 4 months followed by 20 mg/d of simvastatin (n = 2232), who were enrolled in phase Z of the A to Z trial between December 29, 1999, and January 6, 2003. MAIN OUTCOME MEASURE: The primary end point was a composite of cardiovascular death, nonfatal myocardial infarction, readmission for ACS, and stroke. Follow-up was for at least 6 months and up to 24 months. RESULTS: Among the patients in the placebo plus simvastatin group, the median low-density lipoprotein (LDL) cholesterol level achieved while taking placebo was 122 mg/dL (3.16 mmol/L) at 1 month and was 77 mg/dL (1.99 mmol/L) at 8 months while taking 20 mg/d of simvastatin. Among the patients in the simvastatin only group, the median LDL cholesterol level achieved at 1 month while taking 40 mg/d of simvastatin was 68 mg/dL (1.76 mmol/L) and was 63 mg/dL (1.63 mmol/L) at 8 months while taking 80 mg/d of simvastatin. A total of 343 patients (16.7%) in the placebo plus simvastatin group experienced the primary end point compared with 309 (14.4%) in the simvastatin only group (40 mg/80 mg) (hazard ratio [HR], 0.89; 95% confidence interval [CI] 0.76-1.04; P =.14). Cardiovascular death occurred in 109 (5.4%) and 83 (4.1%) patients in the 2 groups (HR, 0.75; 95% CI, 0.57-1.00; P =.05) but no differences were observed in other individual components of the primary end point. No difference was evident during the first 4 months between the groups for the primary end point (HR, 1.01; 95% CI, 0.83-1.25; P =.89), but from 4 months through the end of the study the primary end point was significantly reduced in the simvastatin only group (HR, 0.75; 95% CI, 0.60-0.95; P =.02). Myopathy (creatine kinase >10 times the upper limit of normal associated with muscle symptoms) occurred in 9 patients (0.4%) receiving simvastatin 80 mg/d, in no patients receiving lower doses of simvastatin, and in 1 patient receiving placebo (P =.02). CONCLUSIONS: The trial did not achieve the prespecified end point. However, among patients with ACS, the early initiation of an aggressive simvastatin regimen resulted in a favorable trend toward reduction of major cardiovascular events. PMID- 15337733 TI - Vascular response to angiotensin II in upper body obesity. AB - Upper body obesity is associated with insulin resistance, hypertension, and endothelial dysfunction. We examined forearm vascular function in response to vasodilator (endothelium-dependent and endothelium-independent) and vasoconstrictor stimuli in 8 normotensive, upper body/viscerally obese men with a positive family history of hypertension and 8 age-matched nonobese men. We also measured body composition and insulin regulation of free fatty acid (FFA) and glucose metabolism. Forearm blood flow was measured before and during brachial artery infusions of acetylcholine (Ach), sodium nitroprusside (NTP), and angiotensin II (+/-nitric oxide synthase [NO]) synthase blockade with N(G) monomethyl L-arginine [L-NMMA]). On a separate day, baseline and insulin regulated glucose ([3-3H]glucose) and FFA ([9,10-3H]palmitate) turnover were measured. The vasoconstrictor response to angiotensin II was greater (P<0.05) in obese men than in nonobese men, whereas endothelium-dependent vasodilation was similar. The slope of the angiotensin II dose-response curve correlated significantly with the basal plasma palmitate concentration. Basal and insulin mediated glucose disposal was significantly reduced and FFA turnover significantly increased in viscerally obese men. No differences in endothelium independent vasodilation or relationships between vascular responsivity and palmitate and glucose kinetics or body composition were found. Angiotensin II stimulated forearm vasoconstriction is increased in viscerally obese normotensive men. PMID- 15337734 TI - Modulation of oxidant and antioxidant enzyme expression and function in vascular cells. AB - Pathological conditions that predispose to cardiovascular events, such as hypertension, hypercholesterolemia, and diabetes, are associated with oxidative stress. These observations and further data derived from a plethora of investigations provided accumulating evidence that oxidative stress is decisively involved in the pathogenesis of endothelial dysfunction and atherosclerosis. Several enzymes expressed in vascular tissue contribute to production and efficient degradation of reactive oxygen species, and enhanced activity of oxidant enzymes and/or reduced activity of antioxidant enzymes may cause oxidative stress. Various agonists, pathological conditions, and therapeutic interventions lead to modulated expression and function of oxidant and antioxidant enzymes, including NAD(P)H oxidase, endothelial nitric oxide synthase, xanthine oxidase, myeloperoxidase, superoxide dismutases, catalase, thioredoxin reductase, and glutathione peroxidase. Data from numerous studies underline the importance of dysregulated oxidant and antioxidant enzymes for the development and progression of atherosclerotic disease in animal models and humans. Specific pharmacological modulation of key enzymes involved in the propagation of oxidative stress rather than using direct antioxidants may be an approach to reduce oxygen radical load in the vasculature and subsequent disease progression in humans. This review focuses on the modulation of expression and activity of major antioxidant and oxidant enzymes expressed in vascular cells. PMID- 15337735 TI - Downregulation of matrix metalloproteinases and collagens and suppression of cardiac fibrosis by inhibition of the proteasome. AB - Myocardial remodeling is an adaptive response of the myocardium to several forms of stress culminating in cardiac fibrosis, left ventricular dilation, and loss of contractility. The remodeling processes of the extracellular matrix are controlled by matrix metalloproteinases, which are in turn regulated by growth factors and inflammatory cytokines. The inflammatory transcription factor nuclear factor kappaB has been implicated in the transcriptional regulation of several matrix metalloproteinases. Because activation of nuclear factor kappaB in turn is essentially controlled by the ubiquitin-proteasome system, we investigated the hypothesis that inhibition of the proteasome may prevent activation of matrix metalloproteinases. We demonstrate here that inhibition of the proteasome in rat cardiac fibroblasts suppressed not only expression of matrix metalloproteinases 2 and 9, but also expression of collagen Ialpha1, Ialpha2, and IIIalpha1 as determined by in-gel zymography and real-time reverse transcription-polymerase chain reaction. Moreover, myocardial expression of matrix metalloproteinases and collagens was effectively suppressed by systemic treatment of spontaneously hypertensive rats over 12 weeks with the proteasome inhibitor MG132, which resulted in a marked reduction of cardiac fibrosis (-38%) compared with control animals. We conclude that inhibition of the ubiquitin-proteasome system may provide a new and attractive tool to interfere with collagen and matrix metalloproteinase expression, and therefore might be of possible use in the therapy of myocardial remodeling. PMID- 15337736 TI - Ovariectomy augments hypertension in aging female Dahl salt-sensitive rats. AB - The ovariectomized (OVX) Dahl salt-sensitive (DS) rat fed a low-salt diet is a model of postmenopausal hypertension. In addition to estrogen loss, aging can also contribute to postmenopausal hypertension. We hypothesized that: (1) female DS rats on a low-salt diet become hypertensive with age; (2) ovariectomy accelerates age-dependent hypertension in the DS rat caused by estrogen depletion; and (3) this hypertension correlates with increased type 1 angiotensin receptor (AT1R) number (Bmax). Blood pressure was monitored by telemetry from 3 to 12 months and AT1R Bmax was determined by Scatchard analysis in glomeruli and adrenal cortex. Three groups of DS rats were studied: intact, OVX, and 17beta estradiol-replaced OVX (OVX+E). In intact rats, aging to 12 months resulted in hypertension (159+/-6 mm Hg) and an 82% decrease in estrogen. Blood pressure in OVX was significantly higher than OVX+E through 12 months of age (173+/-4 versus 150+/-8 mm Hg). At 4 months, OVX increased AT1R Bmax compared with intact and OVX+E in both glomeruli and adrenal cortex. Aging also increased AT1R Bmax in these tissues in intact rats. In summary, female DS rats fed a low-salt diet have hypertension develop with age, that is accelerated by OVX and attenuated by estrogen replacement. Concurrently, AT1Rs are upregulated by age and OVX, which is prevented by estrogen replacement. This study suggests that an increased activity of the renin angiotensin system contributes to the development of hypertension, and estrogen protects against this process. PMID- 15337737 TI - Targeting of the renin-Angiotensin system as an adjunct to estrogen replacement therapy. PMID- 15337738 TI - Enhanced rate of cleavage at Arg-306 and Arg-506 in coagulation factor Va by Gla domain-mutated human-activated protein C. AB - A Gla domain-mutated protein C variant, QGNSEDY, modified at positions 10-12, 23, 32-33, and 44, having enhanced affinity for negatively charged phospholipid and increased anticoagulant potential, was used to elucidate the importance of the interaction between the Gla domain and the phospholipid for the ability of activated protein C (APC) to inactivate factor Va (FVa). FVa degradation by wild type (WT)-APC and QGNSEDY-APC yielded similar fragments on Western blotting; QGNSEDY-APC was, however, considerably more efficient. The kinetic parameters for individual APC-mediated cleavages in FVa, i.e. at Arg-306 and Arg-506, were investigated at high and low phospholipid concentrations in the presence and absence of protein S. FVa variants 306Q679Q and 506Q679Q, which can only be cleaved at Arg-506 and Arg-306, respectively, were used. In the absence of protein S, QGNSEDY-APC was 17.8- and 4-fold more efficient than WT-APC in cleaving at Arg-306 and Arg-506, respectively, at high phospholipid. Similar values were obtained at low phospholipid. In the presence of protein S, QGNSEDYAPC was 6.8- and 3.2-fold more active than WT-APC in cleaving at Arg-306 and Arg-506, respectively, at high phospholipid. At low phospholipid, the corresponding values were 14- and 6.5-fold. In conclusion, the modification of the Gla domain in QGNSEDY-APC yielded increased rates of cleavage at both sites in FVa, the increase being particularly pronounced for the Arg-306 site in the absence of protein S. The results obtained with QGNSEDY-APC provide insights into the importance of the APC-phospholipid interaction for the APC-mediated cleavages at Arg-306 and Arg-506 in FVa. PMID- 15337739 TI - The RGS14 GoLoco domain discriminates among Galphai isoforms. AB - Regulators of G protein signaling (RGS) modulate G protein activity by functioning as GTPase-activating proteins (GAPs) for alpha-subunits of heterotrimeric G proteins. RGS14 regulates G protein nucleotide exchange and hydrolysis by acting as a GAP through its RGS domain and as a guanine nucleotide dissociation inhibitor (GDI) through its GoLoco motif. RGS14 exerts GDI activity on Galphai1, but not Galphao. Selective interactions are mediated by contacts between the alphaA and alphaB helices of the Galphai1 helical domain and the GoLoco C terminus (Kimple, R. J., Kimple, M. E., Betts, L., Sondek, J., and Siderovski, D. P. (2002) Nature 416, 878-881). Three isoforms of Galphai exist in mammalian cells. In this study, we tested whether all three isoforms were subject to RGS14 GDI activity. We found that RGS14 inhibits guanine nucleotide exchange on Galphai1 and Galphai3 could, but not Galphai2. Galphai2 be rendered sensitive to RGS14 GDI activity by replacement of residues within the alpha-helical domain. In addition to the contact residues in the alphaA and alphaB helices previously identified, we found that the alphaA/alphaB and alphaB/alphaC loops are important determinants of Galphai selectivity. The striking selectivity observed for RGS14 GDI activity in vitro points to Galphai1 and Galphai3 as the likely targets of RGS14-GoLoco regulation in vivo. PMID- 15337740 TI - Statin induction of liver fatty acid-binding protein (L-FABP) gene expression is peroxisome proliferator-activated receptor-alpha-dependent. AB - Statins are drugs widely used in humans to treat hypercholesterolemia. Statins act by inhibiting cholesterol synthesis resulting in the activation of the transcription factor sterol-responsive element-binding protein-2 that controls the expression of genes involved in cholesterol homeostasis. Statin therapy also decreases plasma triglyceride and non-esterified fatty acid levels, but the mechanism behind this effect remains more elusive. Liver fatty acid-binding protein (L-FABP) plays a role in the influx of long-chain fatty acids into hepatocytes. Here we show that L-FABP is a target for statins. In rat hepatocytes, simvastatin treatment induced L-FABP mRNA levels in a dose-dependent manner. Moreover, L-FABP promoter activity was induced by statin treatment. Progressive 5'-deletion analysis revealed that the peroxisome proliferator activated receptor (PPAR)-responsive element located at position -67/-55 was responsible for the statin-mediated transactivation of the rat L-FABP promoter. Moreover, treatment with simvastatin and the PPARalpha agonist Wy14,649 resulted in a synergistic induction of L-FABP expression (mRNA and protein) in rat Fao hepatoma cells. This effect was also observed in vivo in wild-type mice but not in PPARalpha-null animals demonstrating the direct implication of PPARalpha in L FABP regulation by statin treatment. Statin treatment resulted in a rise in PPARalpha mRNA levels both in vitro and in vivo and activated the mouse PPARalpha promoter in a reporter assay. Altogether, these data demonstrate that L-FABP expression is up-regulated by statins through a mechanism involving PPARalpha. Moreover, PPARalpha might be a statin target gene. These effects might contribute to the triglyceride/non-esterified fatty acid-lowering properties of statins. PMID- 15337741 TI - Monooxygenase X, a member of the copper-dependent monooxygenase family localized to the endoplasmic reticulum. AB - Based on sequence comparisons, MOX (monooxygenase X), is a member of the copper monooxygenase family that includes dopamine beta-monooxygenase (DBM) and peptidylglycine alpha-hydroxylating monooxygenase (PHM). MOX has all of the residues expected to be critical for copper binding, and its cysteine residues can yield the intramolecular disulfide bond pattern observed in DBM. Although DBM and PHM function within the lumen of the secretory pathway, the published sequence for human MOX lacks a signal sequence, suggesting that it does not enter this compartment. We identified an upstream exon that encodes the signal sequence of human MOX. A retained intron yields minor amounts of transcript encoding MOX without a signal sequence. MOX transcripts are widely expressed, with the highest levels in the salivary gland and ovary and moderate levels in brain, pituitary, and heart. Despite the presence of a signal sequence, exogenous MOX is not secreted, and it localizes throughout the endoplasmic reticulum in both endocrine or nonendocrine cells. Neither appending green fluorescent protein to its C terminus nor deleting the hydrophobic domain near its C terminus facilitates secretion of MOX. MOX is N-glycosylated, is tightly membrane-associated, and forms oligomers that are not disulfide-linked. Based on its sequence and localization, MOX is predicted to hydroxylate a hydrophobic substrate in the endoplasmic reticulum. PMID- 15337742 TI - SUMO-1 modification activated GATA4-dependent cardiogenic gene activity. AB - Sumoylation, an important posttranslational modification process, is involved in regulating bioactivities of diverse proteins. GATA4, a cardiac-enriched dual zinc finger transcription factor, plays a critical role in regulating cardiac specified genes. Here, we demonstrated that GATA4 is sumoylated by small ubiquitin-like modifier-1 (SUMO-1), which resulted in enhanced GATA4 transcriptional activity. We further revealed that lysine 366 of GATA4 constituted a major sumyolation site. Conversion of lysine 366 to arginine 366 resulted in reduced GATA4 nuclear occupation, suggesting that SUMO modification may also modulate GATA4 nuclear localization. In GATA4 sumoylation, PIAS1 served as an E3 ligase and positively modulated GATA4 transactivation via its RING finger domain. In the presence of SUMO-1 and/or PIAS1, GATA4 triggered the activation of cardiogenic genes in pluripotent 10T1/2 fibroblast. GATA4 was identified as a SUMO-1-targeted transcription factor and together with PIAS1 was shown to be a potent regulator of cardiac gene activity. PMID- 15337743 TI - Latency and substrate binding globally reduce solvent accessibility of plasminogen activator inhibitor type 1 (PAI-1). An adaptation of PAI-1 conformer crystal structures by hydrogen-deuterium exchange. AB - Plasminogen activator inhibitor type 1 (PAI-1) plays key regulatory roles in fibrinolysis, cell migration, and tissue remodeling. A regulatory protein without known catalytic activity, PAI-1 modulates plasminogen activators through protein protein interactions. Although global conformational alterations that occur in PAI-1 determine its regulatory activity, comprehensive assessments of concurrent dynamic, structural, and functional alterations of this critical regulatory protein have not yet been clearly defined. X-ray crystallographic studies have described four distinct PAI-1 conformational states: active, latent, reactive center loop peptide-annealed (RCL-PA), and cleaved mutant. In this study, backbone amide hydrogen-deuterium exchange detected by mass spectrometry was used to characterize dynamic and structural alterations of human PAI-1 (hPAI-1) in relation to its function. hPAI-1 conformers were defined by surface mapping the solvent-accessible sites for strategic secondary structural components of the protein. We observed a global protection from solvent for a majority of peptides in the latent conformer relative to the active conformer. Significant differences were observed in the RCL, helix A, helix D, and sheet 1C, and these regions were markedly more dynamic or solvent-exposed in the active conformation. The RCL-PA form adopts an intermediate conformational state between the active and the latent conformers. Our results demonstrate that the most dynamic regions of PAI-1 (the RCL, helices D and A, and sheet 5A) are flexible in the transition toward latency. They also show that the dynamic surface structures of the active, latent, and peptide-annealed conformers of PAI-1 are underestimated by theoretical solvent accessibility calculations derived from crystallographic data. PMID- 15337744 TI - Evidence for ligand-independent transcriptional activation of the human estrogen related receptor alpha (ERRalpha): crystal structure of ERRalpha ligand binding domain in complex with peroxisome proliferator-activated receptor coactivator 1alpha. AB - The crystal structure of the ligand binding domain (LBD) of the estrogen-related receptor alpha (ERRalpha, NR3B1) complexed with a coactivator peptide from peroxisome proliferator-activated receptor coactivator-1alpha (PGC-1alpha) reveals a transcriptionally active conformation in the absence of a ligand. This is the first x-ray structure of ERRalpha LBD, solved to a resolution of 2.5 A, and the first structure of a PGC-1alpha complex. The putative ligand binding pocket (LBP) of ERRalpha is almost completely occupied by side chains, in particular with the bulky side chain of Phe328 (corresponding to Ala272 in ERRgamma and Ala350 in estrogen receptor alpha). Therefore, a ligand of a size equivalent to more than approximately 4 carbon atoms could only bind in the LBP, if ERRalpha would undergo a major conformational change (in particular the ligand would displace H12 from its agonist position). The x-ray structure thus provides strong evidence for ligand-independent transcriptional activation by ERRalpha. The interactions of PGC-1alpha with ERRalpha also reveal for the first time the atomic details of how a coactivator peptide containing an inverted LXXLL motif (namely a LLXYL motif) binds to a LBD. In addition, we show that a PGC-1alpha peptide containing this nuclear box motif from the L3 site binds ERRalpha LBD with a higher affinity than a peptide containing a steroid receptor coactivator-1 motif and that the affinity is further enhanced when all three leucine-rich regions of PGC-1alpha are present. PMID- 15337745 TI - Genetic dissection of the phospholipid hydroperoxidase activity of yeast gpx3 reveals its functional importance. AB - Saccharomyces cerevisiae expresses multiple phospholipid hydroperoxide glutathione peroxidase (PHGPx)-like proteins in the absence of a classical glutathione peroxidase (cGPx), providing a unique system for dissecting the roles of these enzymes in vivo. The Gpx3 (Orp1/PHGpx3) protein transduces the hydroperoxide signal to the transcription factor Yap1, a function that could account for most GPX-dependent phenotypes. To test this hypothesis and ascertain what functions of Gpx3 can be shared by cGPx-like enzymes, we constructed a novel cGPx-like yeast enzyme, cGpx3. We confirmed that the "gap" sequences conserved among cGPxs but absent from aligned PHGPx sequences are the principal cause of the structural and functional differences of these enzymes. Peroxidase activity against a cGPx substrate was high in the cGpx3 construct, which was multimeric and had a peroxidase catalytic mechanism distinct from Gpx3; but cGpx3 was defective for phospholipid hydroperoxidase and signaling activities. cGpx3 did not complement the sensitivity to lipid peroxidation of a gpxDelta mutant, and the resistance to lipid peroxidation conferred by Gpx3 was independent of Yap1, establishing a functional role for Gpx3 phospholipid hydroperoxidase activity. Using the comparison between cGpx3 and Gpx3 in conjunction with other constructs to probe lipid peroxidation as a toxicity mechanism, we also ascertained that lipid peroxidation-dependent processes are a principal cause of cellular cadmium toxicity. The results demonstrate that phospholipid hydroperoxidase and Yap1 mediated signaling activities of Gpx3 have independent functional roles, although both functions depend on the absence of cGPx-like subunit interaction sites, and the results resolve more clearly the potential drivers of the differential selective evolution of GPx-like enzymes. PMID- 15337746 TI - Rhodopsin signaling and organization in heterozygote rhodopsin knockout mice. AB - Rhodopsin (Rho) resides within internal membrane structures called disc membranes that are found in the rod outer segments (ROS) of photoreceptors in the retina. Rho expression is essential for formation of ROS, which are absent in knockout Rho-/- mice. ROS of mice heterozygous for the Rho gene deletion (Rho+/-) may have a lower Rho density than wild type (WT) membranes, or the ROS structure may be reduced in size due to lower Rho expression. Here, we present evidence that the smaller volume of ROS from heterozygous mice is most likely responsible for observed electrophysiological response differences. In Rho+/- mice as compared with age-matched WT mice, the length of ROS was shorter by 30-40%, and the average diameter of ROS was reduced by approximately 20%, as demonstrated by transmission and scanning electron microscopy. Together, the reduction of the volume of ROS was approximately 60% in Rho+/- mice. Rho content in the eyes was reduced by approximately 43% and 11-cis-retinal content in the eye was reduced by approximately 38%, as determined by UV-visible spectroscopy and retinoid analysis, respectively. Transmission electron microscopy of negatively stained disc membranes from Rho+/- mice indicated a typical morphology apart from the reduced size of disc diameter. Power spectra calculated from disc membrane regions on such electron micrographs displayed a diffuse ring at approximately 4.5 nm(-1), indicating paracrystallinity of Rho. Atomic force microscopy of WT and Rho+/- disc membranes revealed, in both cases, Rho organized in paracrystalline and raftlike structures. From these data, we conclude that the differences in physiological responses measured in WT and Rho+/- mice are due to structural changes of the whole ROS and not due to a lower density of Rho. PMID- 15337747 TI - Regulation of cGMP-dependent protein kinase expression by soluble guanylyl cyclase in vascular smooth muscle cells. AB - Vascular smooth muscle cells (VSMC) undergo many phenotypic changes when placed in culture. Several studies have shown that the levels of expression of soluble guanylyl cyclase (sGC) or cGMP-dependent protein kinase (PKG) are altered in cultured VSMC. In this study the mechanisms involved in the coordinated expression of sGC and PKG were examined. Pro-inflammatory cytokines that increase the expression of type II NO synthase (inducible NO synthase, or iNOS) decreased PKG expression in freshly isolated, non-passaged bovine aortic SMC. However, in several passaged VSMC lines (i.e. bovine aortic SMC, human aortic SMC, and A7r5 cells), PKG protein expression was not suppressed by cytokines or NO. sGC was highly expressed in non-passaged bovine aortic SMC but not in passaged cell lines. Restoration of expression of sGC to passaged bovine SMC using adenovirus encoding the alpha1 and beta1 subunits of sGC restored the capacity of the cells to increase cGMP in response to NO. Furthermore, treatment of these sGC transduced cells with NO donors for 48 h resulted in decreased PKG protein expression. In contrast, passaged rat aortic SMC expressed high levels of NO responsive sGC but demonstrated reduced expression of PKG. Adenovirus-mediated expression of the PKG catalytically active domain in rat aortic SMC caused a reduction in the expression of sGC in these cells. These results suggest that there is a mechanism for the coordinated expression of sGC and PKG in VSMC and that prolonged activation of sGC down-regulates PKG expression. Likewise, the loss of PKG expression appears to increase sGC expression. These effects may be an adaptive mechanism allowing growth and survival of VSMC in vitro. PMID- 15337748 TI - Identification and characterization of the C3 binding domain of the Staphylococcus aureus extracellular fibrinogen-binding protein (Efb). AB - The secreted Staphylococcus aureus extracellular fibrinogen-binding protein (Efb) is a virulence factor that binds to both the complement component C3b and fibrinogen. Our laboratory previously reported that by binding to C3b, Efb inhibited complement activation and blocked opsonophagocytosis. We have now located the Efb binding domain in C3b to the C3d fragment and determined a disassociation constant (Kd) of 0.24 microM for the Efb-C3d binding using intrinsic fluorescence quenching assays. Using truncated, recombinant forms of Efb, we also demonstrate that the C3b binding region of Efb is located within the C terminus, in contrast to the fibrinogen binding domains that are located at the N-terminal end of the protein. Enzyme-linked immunosorbent assay-type binding assays demonstrated that recombinant Efb could bind to both C3b and fibrinogen simultaneously, forming a trimolecular complex and that the C-terminal region of Efb could inhibit complement activity in vitro. In addition, secondary structure analysis using circular dichroism spectroscopy revealed that the C-terminal, C3b binding region of Efb is composed primarily of alpha-helices, suggesting that this domain of Efb represents a novel type of C3b-binding protein. PMID- 15337749 TI - Restoration of transport activity by co-expression of human reduced folate carrier half-molecules in transport-impaired K562 cells: localization of a substrate binding domain to transmembrane domains 7-12. AB - Reduced folates such as 5-methyl tetrahydrofolate and classical antifolates such as methotrexate are actively transported into mammalian cells by the reduced folate carrier (RFC). RFC is characterized by 12 stretches of mostly hydrophobic, alpha-helix-promoting amino acids, internally oriented N and C termini, and a large central linker connecting transmembrane domains (TMDs) 1-6 and 7-12. Previous studies showed that deletion of the majority of the central loop domain between TMDs 6 and 7 abolished transport, but this segment could be replaced with mostly non-homologous sequence from the SLC19A2 thiamine transporter to restore transport function. In this report, we expressed RFC from separate TMD1-6 and TMD7-12 RFC half-molecule constructs, each with a unique epitope tag, in RFC-null K562 cells to restore transport activity. Restored transport exhibited characteristic transport kinetics for methotrexate, a capacity for trans stimulation by pretreatment with leucovorin, and inhibition by N hydroxysuccinimide methotrexate, a documented affinity inhibitor of RFC. The TMD1 6 half-molecule migrated on SDS gels as a 38-58 kDa glycosylated species and was converted to 27 kDa by N-glycosidase F or tunicamycin treatments. The 40 kDa TMD7 12 half-molecule was unaffected by these treatments. Using transfected cells expressing both TMDs 1-6 and TMDs 7-12 as separate polypeptides, the TMD7-12 half molecule was covalently radiolabeled with N-hydroxysuccinimide [(3)H]methotrexate. No radioactivity was incorporated into the TMD1-6 half molecule. Digestion with endoproteinase GluC decreased the size of the radiolabeled 40 kDa TMD7-12 polypeptide to approximately 20 kDa. Our results demonstrate that a functional RFC can be reconstituted with RFC half-molecules and localize a critical substrate binding domain to within TMDs 7-12. PMID- 15337750 TI - The amino-terminal region of toll-like receptor 4 is essential for binding to MD 2 and receptor translocation to the cell surface. AB - Toll-like receptor 4 (TLR4) and MD-2 are pivotal components that elicit inflammatory responses to lipopolysaccharide (LPS). They have been shown to form a physical complex on the cell surface that responds directly to LPS. However, the functional region of TLR4 required for association with MD-2 and LPS responsiveness is poorly understood. To identify the region of TLR4, we created a series of mutants with deletions in the extracellular domain and examined their activities in human embryonic kidney 293 cells. A mutant with a 317-amino acid deletion from the membrane proximal region of TLR4 was capable of associating with MD-2, while only a 9-amino acid truncation of the N terminus severely impaired the interaction. The association between the two molecules was well correlated with TLR4 maturation into an endoglycosidase H-resistant form and the cell surface expression. Mouse MD-2 bound to human TLR4, but its activity to facilitate the cell surface expression of TLR4 and confer LPS responsiveness was much weaker than that of human MD-2, indicating species specificity. A chimeric receptor composed of the N-terminal region of human TLR4 and the adjacent region of mouse TLR4 showed preference for human MD-2 in its transport to the cell surface and responsiveness to LPS. Taken together, the N-terminal region of TLR4 is essential for association with MD-2, which is required for the cell surface expression and hence the responsiveness to LPS. PMID- 15337751 TI - Impact of engagement of FcepsilonRI and CC chemokine receptor 1 on mast cell activation and motility. AB - CC chemokines participate in the recruitment and activation of immune cells through CC chemokine receptors (CCRs). Here, we report that cross-talk between CCR1-mediated signaling pathway and FcepsilonRI-mediated signaling pathway affects degranulation positively but affects chemotaxis of mast cells adversely. Costimulation via FcepsilonRI engagement with IgE/antigen and CCR1 engagement with recombinant human CCL3 synergistically enhanced degranulation in rat basophilic leukemia-2H3 cells expressing human CCR1 (RBL-CCR1). Interestingly, FcepsilonRI engagement inhibited CCL3-mediated chemotaxis and membrane ruffling of RBL-CCR1 cells. Small GTP-binding proteins of the Rho family, Rac, Cdc42, and Rho control chemotaxis by mediating the reorganization of the actin cytoskeleton. Both a Rho inhibitor C3 exoenzyme and a Rho kinase (ROCK) inhibitor Y-27632 inhibited chemotaxis of RBL-CCR1 cells toward CCL3, indicating that activation of the Rho/ROCK signaling pathway is required for the CCL3-mediated chemotaxis of the cells. Costimulation with IgE/antigen and CCL3 enhanced Rac and Cdc42 activation but decreased ROCK activation in RBL-CCR1 cells compared with that in the cells stimulated with CCL3 alone. These results suggest that costimulation via FcepsilonRI and CCR1 engagements induced 1) inhibition of membrane ruffling, 2) decreased ROCK activation, and 3) reciprocal imbalance between Small GTP binding proteins of the Rho family, which result in the inhibition of chemotaxis of RBL-CCR1 cells. The cross-talk between FcepsilonRI-mediated signaling pathway and CCR-mediated signaling pathway would induce optimal activation and arrested chemotaxis of mast cells, thus contributing to allergic inflammation. PMID- 15337752 TI - Initial bubble collapse plays a key role in the transition to elongation in T7 RNA polymerase. AB - RNA polymerases bind to specific sequences in DNA, melt open duplex DNA around the start site, and start transcription within the initially melted bubble. The initially transcribing complex is relatively unstable, releasing short abortive products. After synthesis of a minimal length of RNA (approximately 10-12 bases in the T7 system), RNA polymerases complete the transition to a processive (highly stable) elongation phase and lose the initial promoter contacts. The current study strongly supports a model for T7 RNA polymerase in which initial bubble collapse from position -4 to position +3 is responsible for initiating RNA displacement in the transition process. More specifically, collapse of the bubble from position -4 to position -1 indirectly and energetically facilitates the direct strand invasion offered by collapse at positions +1 to +3. Parallel work shows that promoter release, another key event occurring during this stage of transcription, begins after translocation to position +8 and is largely complete upon translocation to about position +12. The timing of promoter release agrees with the timing of initial bubble collapse determined by our previous fluorescence studies, suggesting that these two events are closely related. PMID- 15337753 TI - Proteomic analysis of proteins associated with lipid droplets of basal and lipolytically stimulated 3T3-L1 adipocytes. AB - Adipocytes hold the body's major energy reserve as triacylglycerols packaged in large lipid droplets. Perilipins, the most abundant proteins on these lipid droplets, play a critical role in facilitating both triacylglycerol storage and hydrolysis. The stimulation of lipolysis by beta-adrenergic agonists triggers rapid phosphorylation of perilipin and translocation of hormone-sensitive lipase to the surfaces of lipid droplets and more gradual fragmentation and dispersion of micro-lipid droplets. Because few lipid droplet-associated proteins have been identified in adipocytes, we isolated lipid droplets from basal and lipolytically stimulated 3T3-L1 adipocytes and identified the component proteins by mass spectrometry. Structural proteins identified in both preparations include perilipin, S3-12, vimentin, and TIP47; in contrast, adipophilin, caveolin-1, and tubulin selectively localized to droplets in lipolytically stimulated cells. Lipid metabolic enzymes identified in both preparations include hormone-sensitive lipase, lanosterol synthase, NAD(P)-dependent steroid dehydrogenase-like protein, acyl-CoA synthetase, long chain family member (ACSL) 1, and CGI-58. 17-beta Hydroxysteroid dehydrogenase, type 7, was identified only in basal preparations, whereas ACSL3 and 4 and two short-chain reductase/dehydrogenases were identified on droplets from lipolytically stimulated cells. Additionally, both preparations contained FSP27, ribophorin I, EHD2, diaphorase I, and ancient ubiquitous protein. Basal preparations contained CGI-49, whereas lipid droplets from lipolytically stimulated cells contained several Rab GTPases and tumor protein D54. A close association of mitochondria with lipid droplets was suggested by the identification of pyruvate carboxylase, prohibitin, and a subunit of ATP synthase in the preparations. Thus, adipocyte lipid droplets contain specific structural proteins as well as lipid metabolic enzymes; the structural reorganization of lipid droplets in response to the hormonal stimulation of lipolysis is accompanied by increases in the relative mass of several proteins and the recruitment of additional proteins. PMID- 15337754 TI - C-reactive Protein. AB - C-reactive protein (CRP) is a phylogenetically highly conserved plasma protein, with homologs in vertebrates and many invertebrates, that participates in the systemic response to inflammation. Its plasma concentration increases during inflammatory states, a characteristic that has long been employed for clinical purposes. CRP is a pattern recognition molecule, binding to specific molecular configurations that are typically exposed during cell death or found on the surfaces of pathogens. Its rapid increase in synthesis within hours after tissue injury or infection suggests that it contributes to host defense and that it is part of the innate immune response. Recently, an association between minor CRP elevation and future major cardiovascular events has been recognized, leading to the recommendation by the Centers for Disease Control and the American Heart Association that patients at intermediate risk of coronary heart disease might benefit from measurement of CRP. This review will largely focus on our current understanding of the structure of CRP, its ligands, the effector molecules with which it interacts, and its apparent functions. PMID- 15337755 TI - Bacterial acetone carboxylase is a manganese-dependent metalloenzyme. AB - Bacterial acetone carboxylase catalyzes the ATP-dependent carboxylation of acetone to acetoacetate with the concomitant production of AMP and two inorganic phosphates. The importance of manganese in Rhodobacter capsulatus acetone carboxylase has been established through a combination of physiological, biochemical, and spectroscopic studies. Depletion of manganese from the R. capsulatus growth medium resulted in inhibition of acetone-dependent but not malate-dependent cell growth. Under normal growth conditions (0.5 microm Mn2+ in medium), growth with acetone as the carbon source resulted in a 4-fold increase in intracellular protein-bound manganese over malate-grown cells and the appearance of a Mn2+ EPR signal centered at g = 2 that was absent in malate-grown cells. Acetone carboxylase purified from cells grown with 50 microm Mn2+ had a 1.6-fold higher specific activity and 1.9-fold higher manganese content than cells grown with 0.5 microm Mn2+, consistently yielding a stoichiometry of 1.9 manganese/alpha2beta2gamma2 multimer, or 0.95 manganese/alphabetagamma protomer. Manganese in acetone carboxylase was tightly bound and not removed upon dialysis against various metal ion chelators. The addition of acetone to malate-grown cells grown in medium depleted of manganese resulted in the high level synthesis of acetone carboxylase (15-20% soluble protein), which, upon purification, exhibited 7% of the activity and 6% of the manganese content of the enzyme purified from acetone-grown cells. EPR analysis of purified acetone carboxylase indicates the presence of a mononuclear Mn2+ center, with possible spin coupling of two mononuclear sites. The addition of Mg.ATP or Mg.AMP resulted in EPR spectral changes, whereas the addition of acetone, CO2, inorganic phosphate, and acetoacetate did not perturb the EPR. These studies demonstrate that manganese is essential for acetone carboxylation and suggest a role for manganese in nucleotide binding and activation. PMID- 15337756 TI - Distinct ADAM metalloproteinases regulate G protein-coupled receptor-induced cell proliferation and survival. AB - Cross-talk between G protein-coupled receptor (GPCR) and epidermal growth factor receptor (EGFR) signaling systems is widely established in a variety of normal and transformed cell types. Here, we demonstrate that the EGFR transactivation signal requires metalloproteinase cleavage of epidermal growth factor-like growth factor precursors in fibroblasts, ACHN kidney, and TccSup bladder carcinoma cells. Furthermore, we present evidence that blockade of the metalloproteinase disintegrin tumor necrosis factor-alpha-converting enzyme (TACE/ADAM17) by a dominant negative ADAM17 mutant prevents angiotensin II-stimulated pro-HB-EGF cleavage, EGFR activation, and cell proliferation in ACHN tumor cells. Moreover, we found that in TccSup cancer cells, the lysophosphatidic acid-induced transactivation signal is mediated by ADAM15, demonstrating that distinct combinations of growth factor precursors and ADAMs (a disintegrin and metalloproteinases) regulate GPCR-EGFR cross-talk pathways in cell lines derived from urogenital cancer. Our data show further that activation of ADAMs results in discrete cellular responses; whereas GPCR agonists promote activation of the Ras/MAPK pathway and cell proliferation via the EGFR in fibroblasts and ACHN cells, EGFR transactivation pathways regulate activation of the survival mediator Akt/protein kinase B and the susceptibility of fibroblasts and TccSup bladder carcinoma cells to proapoptotic signals such as serum deprivation, death receptor stimulation, and the chemotherapeutic drug doxorubicin. Thus, ADAM15 and -17 function as effectors of GPCR-mediated signaling and define critical characteristics of cancer cells. PMID- 15337757 TI - Murine Wnt-1 with an internal c-myc tag recombinantly produced in Escherichia coli can induce intracellular signaling of the canonical Wnt pathway in eukaryotic cells. AB - Wnt-1 belongs to the Wnt family of secreted glycoproteins inducing an intracellular signaling pathway involved in cell proliferation, differentiation, and pattern formation. The canonical branch is one of three known branches. This is also valid in vitro, and Wnts can be considered beneficial for culturing primary cells from organs, provided Wnts are available and applicable even with cells of different species. It was shown here that internally c-myc-tagged murine Wnt-1 produced in the heterologous host Escherichia coli was appropriate for inducing intracellular signaling of the canonical Wnt pathway in eukaryotic cells via stabilization of cytosolic beta-catenin. The pioneering injection of the protein into the blastocoels of Xenopus laevis embryos led to axis duplication and suppression of head formation. Applying the recombinant murine Wnt-1 to metanephric mesenchyme activated the tubulogenic program. The signal-inducing activity of the recombinant protein was also positively demonstrated in the TOP flash reporter assay. Although Wnts were purified recently from the growth media of stably transfected eukaryotic cell lines, the production of active Wnt proteins in pro- or eukaryotic microorganisms reportedly has never been successful. Here soluble production in E. coli and translocation into the oxidizing environment of the periplasm were achieved. The protein was purified using the internal c-myc tag. The effect on the eukaryotic cells implies that activity was retained. Thus, this approach could make recombinant murine Wnt-1 available as a good starting point for other Wnts needed, for example, for maintaining and differentiating stem cells, organ restoration therapy, and tissue engineering. PMID- 15337758 TI - Fas ligand induces cell-autonomous NF-kappaB activation and interleukin-8 production by a mechanism distinct from that of tumor necrosis factor-alpha. AB - Fas ligand (FasL) has been well characterized as a death factor. However, recent studies revealed that FasL possesses inflammatory activity. Here we found that FasL induces production of the inflammatory chemokine IL-8 without inducing apoptosis in HEK293 cells. Reporter gene assays involving wild-type and mutated IL-8 promoters and NF-kappaB- and AP-1 reporter constructs indicated that an FasL induced NF-kappaB and AP-1 activity are required for maximal promoter activity. FasL induced NF-kappaB activation with slower kinetics than did TNF-alpha, yet this response was cell autonomous and not mediated by secondary paracrine factors. The death domain of Fas, FADD, and caspase-8 were required for NF-kappaB activation by FasL. A dominant-negative mutant of IKKgamma inhibited the FasL induced NF-kappaB activation. However, TRADD and RIP, which are essential for the TNF-alpha-induced NF-kappaB activation, were not involved in the FasL-induced NF kappaB activation. Moreover, CLARP/FLIP inhibited the FasL- but not the TNF-alpha induced NF-kappaB activation. These results show that FasL induces NF-kappaB activation and IL-8 production by a novel mechanism, distinct from that of TNF alpha. In addition, we found that mouse FADD had a dominant-negative effect on the FasL-induced NF-kappaB activation in HEK293 cells, which may indicate a species difference between human and mouse in the FasL-induced NF-kappaB activation. PMID- 15337759 TI - Desnutrin, an adipocyte gene encoding a novel patatin domain-containing protein, is induced by fasting and glucocorticoids: ectopic expression of desnutrin increases triglyceride hydrolysis. AB - We have used rat cDNA microarrays to identify adipocyte-specific genes that could play an important role in adipocyte differentiation or function. Here, we report the cloning and identification of a 2.0-kb mRNA coding for a putative protein that we have designated as desnutrin. The novel gene is expressed predominantly in adipose tissue, and its expression is induced early during 3T3-L1 adipocyte differentiation. Desnutrin mRNA levels were regulated by the nutritional status of animals, being transiently induced during fasting. In vitro desnutrin gene expression was up-regulated by dexamethasone in a dose-dependent manner but not by cAMP, suggesting that glucocorticoids could mediate the increase in desnutrin mRNA levels observed during fasting. Desnutrin mRNA codes for a 486-amino acid putative protein containing a patatin-like domain, characteristic of many plant acyl hydrolases belonging to the patatin family. Confocal microscopy of enhanced green fluorescent protein-tagged desnutrin protein-transfected cells showed that the fusion protein localized in the cytoplasm. Moreover, cells overexpressing desnutrin by transfection showed an increase in triglyceride hydrolysis. Interestingly, we also found that the desnutrin gene expression level was lower in ob/ob and db/db obese mouse models. Overall, our data suggest that the newly identified desnutrin gene codes for an adipocyte protein that may function as a lipase and play a role in the adaptive response to a low energy state, such as fasting, by providing fatty acids to other tissues for oxidation. In addition, decreased expression of desnutrin in obesity models suggests its possible contribution to the pathophysiology of obesity. PMID- 15337760 TI - Vascular endothelial growth factor transcriptional activation is mediated by hypoxia-inducible factor 1alpha, HDM2, and p70S6K1 in response to phosphatidylinositol 3-kinase/AKT signaling. AB - Vascular endothelial growth factor (VEGF) expression is elevated in ovarian and other cancer cells. However, the mechanism that causes the increase in VEGF expression still remains to be elucidated. In this study, we demonstrated that activation of PI3K signaling mediated VEGF protein expression at the transcriptional level through hypoxia-inducible factor 1alpha (HIF-1alpha) expression in human ovarian cancer cells. We found that inhibition of PI3K activity by LY294002 decreased VEGF transcriptional activation and that forced expression of AKT completely reversed the inhibitory effect. HDM2 and p70S6K1 are two downstream targets of AKT that mediate growth factor-induced VEGF transcriptional activation and HIF-1alpha expression. The inhibition of PI3K by LY294002 inhibited p70S6K1 and HDM2 activity in the cells. Forced expression of p70S6K1 or HDM2 reversed LY294002-inhibited VEGF transcriptional activation and HIF-1alpha expression. This study identifies a potential novel mechanism responsible for increased VEGF expression in ovarian cancer cells. It also indicates the important role of VEGF and HIF-1 in ovarian tumorigenesis and angiogenesis, which is mediated by the PI3K/AKT/HDM2 and AKT/p70S6K1 pathways in ovarian cancer cells. PMID- 15337761 TI - Bile acid reduces the secretion of very low density lipoprotein by repressing microsomal triglyceride transfer protein gene expression mediated by hepatocyte nuclear factor-4. AB - Microsomal triglyceride transfer protein (MTP) is involved in the transfer of triglycerides, cholesterol esters, and phospholipids to newly synthesized apolipoprotein (apo) B. It is therefore essential for lipoprotein synthesis and secretion in the liver and the small intestine. Although several recent experiments have revealed the transcriptional regulation of the MTP gene, little has been revealed to date about hepatocyte nuclear factor-4 (HNF-4)-dependent regulation. We here report that the human MTP gene promoter contains a pair of functional responsive elements for HNF-4 and HNF-1, the latter of which is another target gene of HNF-4. Chromatin immunoprecipitation assays provide evidence that endogenous HNF-4 and HNF-1 can bind these elements in chromatin. In Hep G2 cells overexpression of either a dominant negative form of HNF-4 or small interfering RNAs (siRNAs) against HNF-4 dramatically reduces the activities of both the wild type and the HNF-4 site mutant MTP promoter. This suggests that HNF 4 regulates MTP gene expression either directly or indirectly through elevated HNF-1 levels. When Hep G2 cells were cultured with chenodeoxycholic acid (CDCA), a ligand for the farnesoid X receptor (FXR), mRNA levels for MTP and apo B were reduced because of increased expression of the factor small heterodimer partner (SHP), which factor suppresses HNF-4 activities. Chenodeoxycholic acid, but not a synthetic FXR ligand, attenuated expression of HNF-4, bringing about a further suppression of MTP gene expression. Over time the intracellular MTP protein levels and apo B secretion in the culture medium significantly declined. These results indicate that two nuclear receptors, HNF-4 and FXR, are closely involved in MTP gene expression, and the results provide evidence for a novel interaction between bile acids and lipoprotein metabolism. PMID- 15337762 TI - 1,25-dihydroxyvitamin D3 down-regulation of PHEX gene expression is mediated by apparent repression of a 110 kDa transfactor that binds to a polyadenine element in the promoter. AB - The PHEX gene encodes an endopeptidase expressed in osteoblasts that inactivates an uncharacterized peptide hormone, phosphatonin, which suppresses bone mineralization as well as renal phosphate reabsorption and vitamin D bioactivation. We demonstrate that 1alpha-25-dihydroxyvitamin D (1,25(OH)2D3), the, active renal vitamin D metabolite, decreases PHEX mRNA in the rat osteoblastic cell line, UMR-106, as well as in mouse calvaria. Promoter/reporter construct analysis of the murine PHEX gene in transfected UMR-106 cells localized the repressive effect of 1,25(OH)2D3 to the -133 to -74 bp region, and gel mobility shift experiments revealed that 1,25(OH)2D3 treatment of the cells diminished the binding of a nuclear protein(s) to a stretch of 17 adenines from bp -116 to -100 in the proximal PHEX promoter. Either overexpression of a dominant-negative vitamin D receptor (VDR) or deletion of this sequence of 17 A-T base pairs abolished the repressive effect of 1,25(OH)2D3 by attenuating basal promoter activity, indicating that this region mediates the 1,25(OH)2D3 response and is involved in basal transcription. South-western blot analysis and DNA affinity purification show that an unidentified 110 kDa nuclear protein binds to the poly(A) element. Because 1,25(OH)2D3-liganded VDR neither binds to the polyadenine region of the PHEX promoter nor directly influences the association of the 110 kDa transfactor, we conclude that 1,25(OH)2D3 indirectly decreases PHEX expression via VDR-mediated repression (or modification) of this novel transactivator. Thus, we have identified a cis-element required for PHEX gene transcription that participates in negative feedback control of PHEX expression and thereby modulates the actions of phosphatonin. PMID- 15337763 TI - Mitochondrial protein import. Requirement of presequence elements and tom components for precursor binding to the TOM complex. AB - Protein translocation across the outer mitochondrial membrane is mediated by the translocator called the TOM (translocase of the outer mitochondrial membrane) complex. The TOM complex possesses two presequence binding sites on the cytosolic side (the cis site) and on the intermembrane space side (the trans site). Here we analyzed the requirement of presequence elements and subunits of the TOM complex for presequence binding to the cis and trans sites of the TOM complex. The N terminal 14 residues of the presequence of subunit 9 of F(0)-ATPase are required for binding to the trans site. The interaction between the presequence and the cis site is not sufficient to anchor the precursor protein to the TOM complex. Tom7 constitutes or is close to the trans site and has overlapping functions with the C-terminal intermembrane space domain of Tom22 in the mitochondrial protein import. PMID- 15337764 TI - Cellular, biochemical, and genetic analysis of mechanism of small molecule IAP inhibitors. AB - XIAP is member of the IAP family of anti-apoptotic proteins and is known for its ability to bind and suppress caspase family cell death proteases. A phenylurea series of chemical inhibitors of XIAP was recently generated by our laboratories (Schimmer, A. D., Welsh, K., Pinilla, C., Bonneau, M., Wang, Z., Pedersen, I. M., Scott, F. L., Glinsky, G. V., Scudiero, D. A., Sausville, E., Salvesen, G., Nefzi, A., Ostresh, J. M., Houghten, R. A., and Reed, J. C. (2004) Cancer Cell 5, 25-35). We examined the mechanisms of action of these chemical compounds using biochemical, molecular biological, and genetic methods. Active phenylurea-based compounds dissociated effector protease caspase-3 but not initiator protease caspase-9 from XIAP in vitro and restored caspase-3 but not caspase-9 enzymatic activity. When applied to tumor cell lines in culture, active phenylurea-based compounds induced apoptosis in a rapid, concentration-dependent manner, associated with activation of cellular caspases. Apoptosis induced by active phenylurea-based compounds was blocked by chemical inhibitors of caspases, with inhibitors of downstream effector caspases displaying more effective suppression than inhibitors of upstream initiator caspases. Phenylurea-based XIAP antagonists induced apoptosis (defined by annexin V staining) prior to mitochondrial membrane depolarization, in contrast to cytotoxic anticancer drugs. Consistent with these findings, apoptosis induced by phenylurea-based compounds was not altered by genetic alterations in the expression of Bcl-2 family proteins that control mitochondria-dependent cell death pathways, including over-expression of anti apoptotic proteins Bcl-2 or Bcl-X(L) and genetic ablation of pro-apoptotic proteins Bax and Bak. Conversely, conditional over-expression of an active fragment of XIAP or genetic ablation of XIAP expression altered the apoptosis dose-response of the compounds. Altogether, these findings indicate that phenylurea-based XIAP antagonists block interaction of downstream effector caspases with XIAP, thus inducing apoptosis of tumor cell lines through a caspase dependent, Bcl-2/Bax-independent mechanism. PMID- 15337765 TI - The GTP-binding release factor eRF3 as a key mediator coupling translation termination to mRNA decay. AB - GTP is essential for eukaryotic translation termination, where the release factor 3 (eRF3) complexed with eRF1 is involved as the guanine nucleotide-binding protein. In addition, eRF3 regulates the termination-coupled events, eRF3 interacts with poly(A)-binding protein (Pab1) and the surveillance factor Upf1 to mediate normal and nonsense-mediated mRNA decay. However, the roles of GTP binding to eRF3 in these processes remain largely unknown. Here, we showed in yeast that GTP is essentially required for the association of eRF3 with eRF1, but not with Pab1 and Upf1. A mutation in the GTP-binding motifs of eRF3 impairs the eRF1-binding ability without altering the Pab1- or Upf1-binding activity. Interestingly, the mutation causes not only a defect in translation termination but also delay of normal and nonsense-mediated mRNA decay, suggesting that GTP/eRF3-dependent termination exerts its influence on the subsequent mRNA degradation. The termination reaction itself is not sufficient, but eRF3 is essential for triggering mRNA decay. Thus, eRF3 is a key mediator that transduces termination signal to mRNA decay. PMID- 15337766 TI - Leukemia/lymphoma-related factor, a POZ domain-containing transcriptional repressor, interacts with histone deacetylase-1 and inhibits cartilage oligomeric matrix protein gene expression and chondrogenesis. AB - Mutations in the human cartilage oligomeric matrix protein (COMP) gene have been linked to the development of pseudoachondroplasia and multiple epiphyseal dysplasia. We previously cloned the promoter region of the COMP gene and delineated a minimal negative regulatory element (NRE) that is both necessary and sufficient to repress its promoter (Issack, P. S., Fang, C. H., Leslie, M. P., and Di Cesare, P. E. (2000) J. Orthop. Res. 18, 345-350; Issack, P. S., Liu, C. J., Prazak, L., and Di Cesare, P. E. (2004) J. Orthop. Res. 22, 751-758). In this study, a yeast one-hybrid screen for proteins that associate with the NRE led to the identification of the leukemia/lymphoma-related factor (LRF), a transcriptional repressor that contains a POZ (poxvirus zinc finger) domain, as an NRE-binding protein. LRF bound directly to the NRE both in vitro and in living cells. Nine nucleotides (GAGGGTCCC) in the 30-bp NRE are essential for binding to LRF. LRF showed dose-dependent inhibition of COMP-specific reporter gene activity, and exogenous overexpression of LRF repressed COMP gene expression in both rat chondrosarcoma cells and bone morphogenetic protein-2-treated C3H10T1/2 progenitor cells. In addition, LRF also inhibited bone morphogenetic protein-2 induced chondrogenesis in high density micromass cultures of C3H10T1/2 cells, as evidenced by lack of expression of other chondrocytic markers, such as aggrecan and collagen types II, IX, X, and XI, and by Alcian blue staining. LRF associated with histone deacetylase-1 (HDAC1), and experiments utilizing the HDAC inhibitor trichostatin A revealed that LRF-mediated repression requires deacetylase activity. LRF is the first transcription factor found to bind directly to the COMP gene promoter, to recruit HDAC1, and to regulate both COMP gene expression and chondrogenic differentiation. PMID- 15337767 TI - The N-terminal interferon-binding domain (IBiD) homology domain of p300 binds to peptides with homology to the p53 transactivation domain. AB - Two high affinity Ser-20-phospho-LXXLL p53-binding domains of p300 map to the C terminal interferon-binding domain (IBiD) and N-terminal IBiD homology domain (IHD) regions. Purified fractions of a recombinant IHD miniprotein are active in a set of in vitro assays highlighting its affinity to the N-terminal LXXLL domain of p53 including (i) dose-dependent binding to Ser-20-phosphorylated p53 tetramers; (ii) DNA-stimulated binding to p53 tetramers; and (iii) inhibition of MDM2-mediated p53 ubiquitination. The active component of the IHD miniprotein was localized to a 75-amino-acid fragment corresponding to amino acids 401-475 on human p300. This minimal IHD miniprotein can function in vivo as a p53-binding polypeptide in assays including: (i) complex formation with VP16-LXXLL peptide motifs in the two-hybrid assay; (ii) action as a dominant negative inhibitor of p53 from p21 luciferase templates; and (iii) attenuation of endogenous p21 protein levels. Further, we show here that the IRF-1-dependent stabilization and reactivation of p53DeltaPRO protein (LXXLL+/PXXP-) can be neutralized by the minimal IHD miniprotein, suggesting that IHD can bind to the p53 LXXLL domain in vivo. Phage-peptide display to the IHD miniprotein gave rise to an LSQXTFSXLXXLL consensus binding site that displays significant homology to the LXXLL transactivation domain of p53. These data validate the IHD scaffold as an independent LXXLL peptide-binding domain within the p300 protein, complementing the known peptide-binding domains including IBiD, C/H1, and C/H3. PMID- 15337768 TI - The human FA2H gene encodes a fatty acid 2-hydroxylase. AB - 2-Hydroxysphingolipids are a subset of sphingolipids containing 2-hydroxy fatty acids. The 2-hydroxylation occurs during de novo ceramide synthesis and is catalyzed by fatty acid 2-hydroxylase (also known as fatty acid alpha hydroxylase). In mammals, 2-hydroxysphingolipids are present abundantly in brain because the major myelin lipids galactosylceramides and sulfatides contain 2 hydroxy fatty acids. Here we report identification and characterization of a human gene that encodes a fatty acid 2-hydroxylase. Data base searches revealed a human homologue of the yeast ceramide 2-hydroxylase gene (FAH1), which we named FA2H. The FA2H gene encodes a 372-amino acid protein with 36% identity and 46% similarity to yeast Fah1p. The amino acid sequence indicates that FA2H protein contains an N-terminal cytochrome b5 domain and four potential transmembrane domains. FA2H also contains the iron-binding histidine motif conserved among membrane-bound desaturases/hydroxylases. COS7 cells expressing human FA2H contained 3-20-fold higher levels of 2-hydroxyceramides (C16, C18, C24, and C24:1) and 2-hydroxy fatty acids compared with control cells. Microsomal fractions prepared from transfected COS7 cells showed tetracosanoic acid 2 hydroxylase activities in an NADPH- and NADPH: cytochrome P-450 reductase dependent manner. FA2H lacking the N-terminal cytochrome b5 domain had little activity, indicating that this domain is a functional component of this enzyme. Northern blot analysis showed that the FA2H gene is highly expressed in brain and colon tissues. These results demonstrate that the human FA2H gene encodes a fatty acid 2-hydroxylase. FA2H is likely involved in the formation of myelin 2-hydroxy galactosylceramides and -sulfatides. PMID- 15337769 TI - Induction of stem cell factor/c-Kit/slug signal transduction in multidrug resistant malignant mesothelioma cells. AB - Malignant mesothelioma (MM) is strongly resistant to conventional chemotherapy by unclear mechanisms. We and others have previously reported that cytokine- and growth factor-mediated signal transduction is involved in the growth and progression of MM. Here, we identified a pathway that involves stem cell factor (SCF)/c-Kit/Slug in mediating multidrug resistance of MM cells. When we compared gene expression profiles between five MM cells and their multidrug-resistant (MM DX) sublines, we found that MM DX cells expressed both SCF and c-Kit and had higher mRNA levels of Slug. Knockdown of c-Kit or Slug expression with their respective small interfering RNA sensitized MM DX cells to the induction of apoptosis by different chemotherapeutic agents, including doxorubicin, paclitaxel, and vincristine. Transfection of c-Kit in parental MM cells in the presence of SCF up-regulated Slug and increased resistance to the chemotherapeutic agents. Moreover, MM cells expressing Slug showed a similar increased resistance to the chemotherapeutic agents. These results indicate that induction of Slug by autocrine production of SCF and c-Kit activation plays a key role in conferring a broad spectrum chemoresistance on MM cells and reveal a novel signal transduction pathway for pharmacological or genetic intervention of MM patients. PMID- 15337770 TI - Phosphorylation and specific ubiquitin acceptor sites are required for ubiquitination and degradation of the IFNAR1 subunit of type I interferon receptor. AB - Ubiquitination, endocytosis, and lysosomal degradation of the IFNAR1 (interferon alpha receptor 1) subunit of the type I interferon (IFN) receptor is mediated by the SCFbeta-Trcp (Skp1-Cullin1-F-box protein beta transducin repeat-containing protein) E3 ubiquitin ligase in a phosphorylation-dependent manner. In addition, stability of IFNAR1 is regulated by its binding to Tyk2 kinase. Here we characterize the determinants of IFNAR1 ubiquitination and degradation. We found that the integrity of two Ser residues at positions 535 and 539 within the specific destruction motif present in the cytoplasmic tail of IFNAR1 is essential for the ability of IFNAR1 to recruit beta-Trcp as well as to undergo efficient ubiquitination and degradation. Using an antibody that specifically recognizes IFNAR1 phosphorylated on Ser535 we found that IFNAR1 is phosphorylated on this residue in cells. This phosphorylation is promoted by treatment of cells with IFNalpha. Although the cytoplasmic tail of IFNAR1 contains seven Lys residues that could function as potential ubiquitin acceptor sites, we found that only three (Lys501, Lys525, and Lys526), all located proximal to the destruction motif, are essential for ubiquitination and degradation of IFNAR1. Expression of Tyk2 stabilized IFNAR1 in a manner that was dependent neither on its binding to beta-Trcp nor IFNAR1 ubiquitination. We discuss the complexities and specifics of the ubiquitination and degradation of IFNAR1, which is a beta-Trcp substrate that undergoes degradation via a lysosomal pathway. PMID- 15337771 TI - The JEM and the JCB: a synergistic partnership. PMID- 15337772 TI - Yeast actin patches are networks of branched actin filaments. AB - Cortical actin patches are the most prominent actin structure in budding and fission yeast. Patches assemble, move, and disassemble rapidly. We investigated the mechanisms underlying patch actin assembly and motility by studying actin filament ultrastructure within a patch. Actin patches were partially purified from Saccharomyces cerevisiae and examined by negative-stain electron microscopy (EM). To identify patches in the EM, we correlated fluorescence and EM images of GFP-labeled patches. Patches contained a network of actin filaments with branches characteristic of Arp2/3 complex. An average patch contained 85 filaments. The average filament was only 50-nm (20 actin subunits) long, and the filament to branch ratio was 3:1. Patches lacking Sac6/fimbrin were unstable, and patches lacking capping protein were relatively normal. Our results are consistent with Arp2/3 complex-mediated actin polymerization driving yeast actin patch assembly and motility, as described by a variation of the dendritic nucleation model. PMID- 15337773 TI - Pericentrin forms a complex with intraflagellar transport proteins and polycystin 2 and is required for primary cilia assembly. AB - Primary cilia are nonmotile microtubule structures that assemble from basal bodies by a process called intraflagellar transport (IFT) and are associated with several human diseases. Here, we show that the centrosome protein pericentrin (Pcnt) colocalizes with IFT proteins to the base of primary and motile cilia. Immunogold electron microscopy demonstrates that Pcnt is on or near basal bodies at the base of cilia. Pcnt depletion by RNA interference disrupts basal body localization of IFT proteins and the cation channel polycystin-2 (PC2), and inhibits primary cilia assembly in human epithelial cells. Conversely, silencing of IFT20 mislocalizes Pcnt from basal bodies and inhibits primary cilia assembly. Pcnt is found in spermatocyte IFT fractions, and IFT proteins are found in isolated centrosome fractions. Pcnt antibodies coimmunoprecipitate IFT proteins and PC2 from several cell lines and tissues. We conclude that Pcnt, IFTs, and PC2 form a complex in vertebrate cells that is required for assembly of primary cilia and possibly motile cilia and flagella. PMID- 15337774 TI - Membrane insertion of anthrax protective antigen and cytoplasmic delivery of lethal factor occur at different stages of the endocytic pathway. AB - The protective antigen (PA) of anthrax toxin binds to a cell surface receptor, undergoes heptamerization, and binds the enzymatic subunits, the lethal factor (LF) and the edema factor (EF). The resulting complex is then endocytosed. Via mechanisms that depend on the vacuolar ATPase and require membrane insertion of PA, LF and EF are ultimately delivered to the cytoplasm where their targets reside. Here, we show that membrane insertion of PA already occurs in early endosomes, possibly only in the multivesicular regions, but that subsequent delivery of LF to the cytoplasm occurs preferentially later in the endocytic pathway and relies on the dynamics of internal vesicles of multivesicular late endosomes. PMID- 15337775 TI - Kazrin, a novel periplakin-interacting protein associated with desmosomes and the keratinocyte plasma membrane. AB - Periplakin forms part of the scaffold onto which the epidermal cornified envelope is assembled. The NH2-terminal 133 amino acids mediate association with the plasma membrane and bind a novel protein, kazrin. Kazrin is highly conserved and lacks homology to any known protein. There are four alternatively spliced transcripts, encoding three proteins with different NH2 termini. Kazrin is expressed in all layers of stratified squamous epithelia; it becomes membrane associated in the suprabasal layers, coincident with up-regulation of periplakin, and is incorporated into the cornified envelope of cultured keratinocytes. Kazrin colocalizes with periplakin and desmoplakin at desmosomes and with periplakin at the interdesmosomal plasma membrane, but its subcellular distribution is independent of periplakin. On transfection, all three kazrin isoforms have similar subcellular distributions. We conclude that kazrin is a novel component of desmosomes that associates with periplakin. PMID- 15337776 TI - Sonic hedgehog regulates the proliferation, differentiation, and migration of enteric neural crest cells in gut. AB - Enteric neural crest cells (NCCs) migrate and colonize the entire gut and proliferate and differentiate into neurons and glia of the enteric nervous system in vertebrate embryos. We have investigated the mitogenic and morphogenic functions of Sonic hedgehog (Shh) on enteric NCCs in cell and organ culture. Enteric NCCs expressed Shh receptor Patched and transcripts encoding the Shh signal transducer (Gli1). Shh promoted the proliferation and inhibited the differentiation of NCCs. The pro-neurogenic effect of glial cell line--derived neurotrophic factor (GDNF) on NCCs was abolished by Shh. In gut explants, NCCs migrated from the explants onto the adjacent substratum if GDNF was added, whereas addition of Shh abolished this migration. Neuronal differentiation and coalescence of neural crest--derived cells into myenteric plexuses in explants was repressed by the addition of Shh. Our data suggest that Shh controls the proliferation and differentiation of NCCs and modulates the responsiveness of NCCs toward GDNF inductions. PMID- 15337777 TI - Sorting of a nonmuscle tropomyosin to a novel cytoskeletal compartment in skeletal muscle results in muscular dystrophy. AB - Tropomyosin (Tm) is a key component of the actin cytoskeleton and >40 isoforms have been described in mammals. In addition to the isoforms in the sarcomere, we now report the existence of two nonsarcomeric (NS) isoforms in skeletal muscle. These isoforms are excluded from the thin filament of the sarcomere and are localized to a novel Z-line adjacent structure. Immunostained cross sections indicate that one Tm defines a Z-line adjacent structure common to all myofibers, whereas the second Tm defines a spatially distinct structure unique to muscles that undergo chronic or repetitive contractions. When a Tm (Tm3) that is normally absent from muscle was expressed in mice it became associated with the Z-line adjacent structure. These mice display a muscular dystrophy and ragged-red fiber phenotype, suggestive of disruption of the membrane-associated cytoskeletal network. Our findings raise the possibility that mutations in these tropomyosin and these structures may underpin these types of myopathies. PMID- 15337778 TI - Phospholipase C and cofilin are required for carcinoma cell directionality in response to EGF stimulation. AB - The epidermal growth factor (EGF)-induced increase in free barbed ends, resulting in actin polymerization at the leading edge of the lamellipodium in carcinoma cells, occurs as two transients: an early one at 1 min and a late one at 3 min. Our results reveal that phospholipase (PLC) is required for triggering the early barbed end transient. Phosphoinositide-3 kinase selectively regulates the late barbed end transient. Inhibition of PLC inhibits cofilin activity in cells during the early transient, delays the initiation of protrusions, and inhibits the ability of cells to sense a gradient of EGF. Suppression of cofilin, using either small interfering RNA silencing or function-blocking antibodies, selectively inhibits the early transient. Therefore, our results demonstrate that the early PLC and cofilin-dependent barbed end transient is required for the initiation of protrusions and is involved in setting the direction of cell movement in response to EGF. PMID- 15337779 TI - Bend propagation drives central pair rotation in Chlamydomonas reinhardtii flagella. AB - Regulation of motile 9+2 cilia and flagella depends on interactions between radial spokes and a central pair apparatus. Although the central pair rotates during bend propagation in flagella of many organisms and rotation correlates with a twisted central pair structure, propulsive forces for central pair rotation and twist are unknown. Here we compared central pair conformation in straight, quiescent flagella to that in actively beating flagella using wild-type Chlamydomonas reinhardtii and mutants that lack radial spoke heads. Twists occur in quiescent flagella in both the presence and absence of spoke heads, indicating that spoke--central pair interactions are not needed to generate torque for twisting. Central pair orientation in propagating bends was also similar in wild type and spoke head mutant strains, thus orientation is a passive response to bend formation. These results indicate that bend propagation drives central pair rotation and suggest that dynein regulation by central pair--radial spoke interactions involves passive central pair reorientation to changes in bend plane. PMID- 15337780 TI - The raft-associated protein MAL is required for maintenance of proper axon--glia interactions in the central nervous system. AB - The myelin and lymphocyte protein (MAL) is a tetraspan raft-associated proteolipid predominantly expressed by oligodendrocytes and Schwann cells. We show that genetic ablation of mal resulted in cytoplasmic inclusions within compact myelin, paranodal loops that are everted away from the axon, and disorganized transverse bands at the paranode--axon interface in the adult central nervous system. These structural changes were accompanied by a marked reduction of contactin-associated protein/paranodin, neurofascin 155 (NF155), and the potassium channel Kv1.2, whereas nodal clusters of sodium channels were unaltered. Initial formation of paranodal regions appeared normal, but abnormalities became detectable when MAL started to be expressed. Biochemical analysis revealed reduced myelin-associated glycoprotein, myelin basic protein, and NF155 protein levels in myelin and myelin-derived rafts. Our results demonstrate a critical role for MAL in the maintenance of central nervous system paranodes, likely by controlling the trafficking and/or sorting of NF155 and other membrane components in oligodendrocytes. PMID- 15337781 TI - Role of lipid rafts in E-cadherin-- and HGF-R/Met--mediated entry of Listeria monocytogenes into host cells. AB - Listeria monocytogenes uptake by nonphagocytic cells is promoted by the bacterial invasion proteins internalin and InlB, which bind to their host receptors E cadherin and hepatocyte growth factor receptor (HGF-R)/Met, respectively. Here, we present evidence that plasma membrane organization in lipid domains is critical for Listeria uptake. Cholesterol depletion by methyl-beta-cyclodextrin reversibly inhibited Listeria entry. Lipid raft markers, such as glycosylphosphatidylinositol-linked proteins, a myristoylated and palmitoylated peptide and the ganglioside GM1 were recruited at the bacterial entry site. We analyzed which molecular events require membrane cholesterol and found that the presence of E-cadherin in lipid domains was necessary for initial interaction with internalin to promote bacterial entry. In contrast, the initial interaction of InlB with HGF-R did not require membrane cholesterol, whereas downstream signaling leading to F-actin polymerization was cholesterol dependent. Our work, in addition to documenting for the first time the role of lipid rafts in Listeria entry, provides the first evidence that E-cadherin and HGF-R require lipid domain integrity for their full activity. PMID- 15337788 TI - Inhibition of T cell receptor signal transduction by tyrosine kinase-interacting protein of Herpesvirus saimiri. AB - T cells play a central role in orchestrating immunity against pathogens, particularly viruses. Thus, impairing T cell activation is an important strategy employed by viruses to escape host immune control. The tyrosine kinase interacting protein (Tip) of the T lymphotropic Herpesvirus saimiri (HVS) is constitutively present in lipid rafts and interacts with cellular Lck tyrosine kinase and p80 endosomal protein. Here we demonstrate that, due to the sequestration of Lck by HVS Tip, T cell receptor (TCR) stimulation fails to activate ZAP70 tyrosine kinase and to initiate downstream signaling events. TCR zeta chains in Tip-expressing T cells were initially phosphorylated to recruit ZAP70 molecule upon TCR stimulation, but the recruited ZAP70 kinase was not subsequently phosphorylated, resulting in TCR complexes that were stably associated with inactive ZAP70 kinase. Consequently, Tip expression not only markedly inhibited TCR-mediated intracellular signal transduction but also blocked TCR engagement with major histocompatibility complexes on the antigen presenting cells and immunological synapse formation. These results demonstrate that a lymphotropic herpesvirus has evolved a novel mechanism to deregulate T cell activation to disarm host immune surveillance. This process contributes to the establishment and maintenance of viral latency. PMID- 15337789 TI - The same IkappaBalpha mutation in two related individuals leads to completely different clinical syndromes. AB - Both innate and adaptive immune responses are dependent on activation of nuclear factor kappaB (NF-kappaB), induced upon binding of pathogen-associated molecular patterns to Toll-like receptors (TLRs). In murine models, defects in NF-kappaB pathway are often lethal and viable knockout mice have severe immune defects. Similarly, defects in the human NF-kappaB pathway described to date lead to severe clinical disease. Here, we describe a patient with a hyper immunoglobulin M-like immunodeficiency syndrome and ectodermal dysplasia. Monocytes did not produce interleukin 12p40 upon stimulation with various TLR stimuli and nuclear translocation of NF-kappaB was impaired. T cell receptor-mediated proliferation was also impaired. A heterozygous mutation was found at serine 32 in IkappaBalpha. Interestingly, his father has the same mutation but displays complex mosaicism. He does not display features of ectodermal dysplasia and did not suffer from serious infections with the exception of a relapsing Salmonella typhimurium infection. His monocyte function was impaired, whereas T cell function was relatively normal. Consistent with this, his T cells almost exclusively displayed the wild-type allele, whereas both alleles were present in his monocytes. We propose that the T and B cell compartment of the mosaic father arose as a result of selection of wild-type cells and that this underlies the widely different clinical phenotype. PMID- 15337790 TI - Lnk inhibits Tpo-mpl signaling and Tpo-mediated megakaryocytopoiesis. AB - Thrombopoietin (Tpo) is the primary cytokine regulating megakaryocyte development and platelet production. Tpo signaling through its receptor, c-mpl, activates multiple pathways including signal transducer and activator of transcription (STAT)3, STAT5, phosphoinositide 3-kinase-Akt, and p42/44 mitogen-activated protein kinase (MAPK). The adaptor protein Lnk is implicated in cytokine receptor and immunoreceptor signaling. Here, we show that Lnk overexpression negatively regulates Tpo-mediated cell proliferation and endomitosis in hematopoietic cell lines and primary hematopoietic cells. Lnk attenuates Tpo-induced S-phase progression in 32D cells expressing mpl, and Lnk decreases Tpo-dependent megakaryocyte growth in bone marrow (BM)-derived megakaryocyte culture. Consistent with this result, we found that in both BM and spleen, Lnk-deficient mice exhibited increased numbers of megakaryocytes with increased ploidy compared with wild-type mice. In addition, Lnk-deficient megakaryocytes derived from BM and spleen showed enhanced sensitivity to Tpo during culture. The absence of Lnk caused enhanced and prolonged Tpo induction of STAT3, STAT5, Akt, and MAPK signaling pathways in CD41+ megakaryocytes. Furthermore, the Src homology 2 domain of Lnk is essential for Lnk's inhibitory function. In contrast, the conserved tyrosine near the COOH terminus is dispensable and the pleckstrin homology domain of Lnk contributes to, but is not essential for, inhibiting Tpo dependent 32D cell growth or megakaryocyte development. Thus, Lnk negatively modulates mpl signaling pathways and is important for Tpo-mediated megakaryocytopoiesis in vivo. PMID- 15337791 TI - Amphiphysin IIm is required for survival of Chlamydia pneumoniae in macrophages. AB - Macrophages play a critical role in both innate and acquired immunity because of their unique ability to internalize, kill, and degrade bacterial pathogens through the process of phagocytosis. The adaptor protein, amphiphysin IIm, participates in phagocytosis and is transiently associated with early phagosomes. Certain pathogens, including Chlamydia pneumoniae, have evolved mechanisms to subvert macrophage phagosome maturation and, thus, are able to survive within these cells. We report here that, although amphiphysin IIm is usually only transiently associated with the phagosome, it is indefinitely retained on vacuoles containing C. pneumoniae. Under these wild-type conditions, C. pneumoniae do not elicit significant nitric oxide (NO) production and are not killed. Abrogation of amphiphysin IIm function results in C. pneumoniae-induced NO production and in the sterilization of the vacuole. The data suggest that C. pneumoniae retains amphiphysin IIm on the vacuole to survive within the macrophage. PMID- 15337792 TI - Theophylline restores histone deacetylase activity and steroid responses in COPD macrophages. AB - Chronic obstructive pulmonary disease (COPD) is a common chronic inflammatory disease of the lungs with little or no response to glucocorticoids and a high level of oxidative stress. Histone deacetylase (HDAC) activity is reduced in cells of cigarette smokers, and low concentrations of theophylline can increase HDAC activity. We measured the effect of theophylline on HDAC activity and inflammatory gene expression in alveolar macrophages (AM) from patients with COPD. AM from normal smokers showed a decrease in HDAC activity compared with normal control subjects, and this was further reduced in COPD patients (51% decrease, P < 0.01). COPD AMs also showed increased basal release of IL-8 and TNF alpha, which was poorly suppressed by dexamethasone. Theophylline induced a sixfold increase in HDAC activity in COPD AM lysates and significantly enhanced dexamethasone suppression of induced IL-8 release, an effect that was blocked by the HDAC inhibitor trichostatin A. Therefore, theophylline might restore steroid responsiveness in COPD patients. PMID- 15337794 TI - Neoadjuvant chemotherapy followed by extrapleural pneumonectomy in malignant pleural mesothelioma. AB - PURPOSE: To investigate neoadjuvant chemotherapy with cisplatin and gemcitabine followed by extrapleural pneumonectomy with or without radiation therapy in patients with potentially resectable malignant pleural mesothelioma (MPM). PATIENTS AND METHODS: Eligible patients had MPM with clinical stage T1-3, N0-2, M0 disease considered to be completely resectable and a WHO performance status of 0 to 2. Neoadjuvant chemotherapy consisted of three cycles of cisplatin 80 mg/m2 on day 1 and gemcitabine 1,000 mg/m2 on days 1, 8, and 15, given every 28 days. Surgery had to consist of a complete extrapleural pneumonectomy, including resection of pericardium and diaphragm. Postoperative radiotherapy was to be considered for all patients. RESULTS: Nineteen patients with MPM were included in this pilot study. According to the European Organization for Research and Treatment of Cancer prognostic score, two patients were in the good prognosis group, and 17 patients were in the poor prognosis group. The response rate to neoadjuvant chemotherapy was 32%. The major toxicity was thrombocytopenia. Extrapleural pneumonectomy was performed in 16 patients with no perioperative mortality. Major surgical complications occurred in six patients, and all were treated successfully. Thirteen patients received postoperative radiotherapy. The median survival time was 23 months. Two patients remain alive and free of disease 41 and 38 months after initiation of therapy. CONCLUSION: For patients with potentially operable MPM, the availability of active and well-tolerated chemotherapy regimens, the fact that extrapleural pneumonectomy can be safely performed after neoadjuvant chemotherapy in an experienced center, and the promising results regarding survival in our pilot study warrant further investigation of the role of neoadjuvant chemotherapy in a multimodality strategy. PMID- 15337793 TI - Activated polymorphonuclear leukocytes rapidly synthesize retinoic acid receptor alpha: a mechanism for translational control of transcriptional events. AB - In addition to releasing preformed granular proteins, polymorphonuclear leukocytes (PMNs) synthesize chemokines and other factors under transcriptional control. Here we demonstrate that PMNs express an inducible transcriptional modulator by signal-dependent activation of specialized mechanisms that regulate messenger RNA (mRNA) translation. HL-60 myelocytic cells differentiated to surrogate PMNs respond to activation by platelet activating factor by initiating translation and with appearance of specific mRNA transcripts in polyribosomes. cDNA array analysis of the polyribosome fraction demonstrated that retinoic acid receptor (RAR)-alpha, a transcription factor that controls the expression of multiple genes, is one of the polyribosome-associated transcripts. Quiescent surrogate HL60 PMNs and primary human PMNs contain constitutive message for RAR alpha but little or no protein. RAR-alpha protein is rapidly synthesized in response to platelet activating factor under the control of a specialized translational regulator, mammalian target of rapamycin, and is blocked by the therapeutic macrolide rapamycin, events consistent with features of the 5' untranslated region of the transcript. Newly synthesized RAR-alpha modulates production of interleukin-8. Rapid expression of a transcription factor under translational control is a previously unrecognized mechanism in human PMNs that indicates unexpected diversity in gene regulation in this critical innate immune effector cell. PMID- 15337795 TI - Use of surgery among elderly patients with stage IV colorectal cancer. AB - PURPOSE: The role of surgery to remove the primary tumor among patients with stage IV colorectal cancer (CRC) is controversial. The purpose of this study was to evaluate surgical practice patterns for patients > or = 65 years of age with stage IV CRC in a US population-based cohort. PATIENTS AND METHODS: We used the Surveillance, Epidemiology, and End Results-Medicare-linked database to evaluate the patterns of cancer treatment for 9,011 Medicare beneficiaries presenting with stage IV CRC from 1991 to 1999. Patients were categorized according to whether they had primary-cancer-directed surgery (CDS) or no CDS within 4 months of diagnosis. The use of other treatment modalities, including metastasectomy, chemotherapy, and radiation, was evaluated in relationship to whether patients belonged to the CDS or no CDS group. RESULTS: Seventy-two percent (6,469 of 9,011) of patients received CDS, and their 30-day postoperative mortality was 10%. Patients with left-sided or rectal lesions, patients older than age 75 years, blacks, and those of lower socioeconomic status were less likely to undergo CDS; but even among those older than age 75, the CDS rate was 69% (3,378 of 4,909). In contrast, chemotherapy use was less common (47% for patients who had CDS and 31% for those who did not). Metastasectomy was rare; only 3.9% of patients underwent these operations at any point from diagnosis to death. CONCLUSION: Palliative resection of the primary tumor is often performed for elderly US patients with stage IV colorectal cancer. This practice pattern merits re-evaluation, given the improvement in the efficacy of systemic chemotherapy. PMID- 15337796 TI - How accurate is clinician reporting of chemotherapy adverse effects? A comparison with patient-reported symptoms from the Quality-of-Life Questionnaire C30. AB - PURPOSE: Adverse events in chemotherapy clinical trials are assessed and reported by clinicians, yet clinician accuracy in assessing symptoms has been questioned. We compared patient reporting of eight symptoms using a validated instrument, the European Organization for the Research and Treatment of Cancer Quality-of-Life Questionnaire C30 (QLQ-C30 or QLQ) with physicians' reporting of the same symptoms in the study's adverse events log. PATIENTS AND METHODS: Thirty-seven men with metastatic, androgen-independent prostate cancer enrolled onto a phase II trial of weekly calcitriol and docetaxel completed the QLQ every 4 weeks for up to 28 weeks. A patient-reported symptom was defined as an increase in a QLQ symptom score by at least 10 points (0 to 100 scale), sustained for at least 4 weeks. A physician-reported symptom was considered present if it was ever documented in the adverse event log. RESULTS: Forty-nine (new or worsened) symptoms were detected by both physician and QLQ, 48 symptoms were detected by the physician alone, and 55 symptoms were detected by the QLQ alone. They agreed on the absence of a symptom in 102 instances of 254 possible opportunities. Their uncorrected agreement was 59.4%, but Cohen's kappa, a coefficient of agreement that corrects for chance, was 0.15, indicating only slight agreement. Using the QLQ as the standard, overall physician sensitivity and specificity was 47% and 68%, respectively, although it varied considerably among symptoms. CONCLUSION: Even in a tightly controlled clinical trial, physician reporting was neither sensitive nor specific in detecting common chemotherapy adverse effects. Tools for collecting patient-reported adverse event data in chemotherapy clinical trials should be developed. PMID- 15337797 TI - Nuclear expression of BCL10 or nuclear factor kappa B predicts Helicobacter pylori-independent status of early-stage, high-grade gastric mucosa-associated lymphoid tissue lymphomas. AB - PURPOSE: A high percentage of early-stage, high-grade gastric mucosa-associated lymphoid tissue (MALT) lymphomas remain Helicobacter pylori dependent. t(11;18)(q21;q21), a genetic aberration highly predictive of H. pylori independent status in low-grade gastric MALT lymphoma, is rarely detected in its high-grade counterpart. This study examined whether nuclear expression of BCL10 or nuclear factor kappa B (NF-kappaB) is useful in predicting H. pylori independent status in patients with stage IE high-grade gastric MALT lymphomas. PATIENTS AND METHODS: Twenty-two patients who had participated in a prospective study of H. pylori eradication for stage IE high-grade gastric MALT lymphomas were studied. The expression of BCL10 and NF-kappaB in pretreatment paraffin embedded lymphoma tissues was evaluated by immunohistochemistry and confocal immunofluorescence microscopy. The presence of t(11;18)(q21;q21) was identified by a multiplex reverse transcriptase polymerase chain reaction of the API2-MALT1 chimeric transcript. RESULTS: Aberrant nuclear expression of BCL10 was detected in seven (87.5%) of eight H. pylori-independent and in none of 14 H. pylori dependent high-grade gastric MALT lymphomas (P <.001). All seven patients with nuclear BCL10 expression had nuclear expression of NF-kappaB, compared with only two of 15 patients without nuclear BCL10 expression (P =.002). As a single variable, the frequency of nuclear expression of NF-kappaB was also significantly higher in H. pylori-independent tumors than in H. pylori-dependent tumors (seven of eight [87.5%] v two of 15 [12.3%]; P =.002). The API2-MALT1 fusion transcript was detected in only one (12.5%) of eight H. pylori-independent lymphomas. CONCLUSION: Nuclear expression of BCL10 or NF-kappaB is highly predictive of H. pylori-independent status in high-grade gastric MALT lymphoma, and coexpression of these two markers in the nuclei is frequent. PMID- 15337798 TI - Clinicopathologic significance and prognostic value of chromosomal imbalances in diffuse large B-cell lymphomas. AB - PURPOSE: To determine the clinicopathologic significance and prognostic value of chromosomal imbalances in diffuse large B-cell lymphomas (DLBCL). PATIENTS AND METHODS: We have examined 64 tumors at diagnosis using comparative genomic hybridization and real-time quantitative polymerase chain reaction (PCR), single stranded conformational polymorphism, and DNA sequencing for the analysis of several potential target genes. RESULTS: The most recurrent alterations were gains of 18q (20%), Xq (15%), 2p, 7q, and 12p (14%), and losses of 6q and 17p (14%). Frequent high-level DNA amplifications were detected at 2p13-p16 and 18q21 loci. Real-time quantitative PCR detected REL and BCL11A gene amplifications in the nine patients with gains at 2p13-p16 and only in one additional patient with normal chromosome 2. Similarly, the BCL-2 gene was amplified in the 12 tumors with gains of 18q21 but in none of 39 patients with normal 18q profile. p53 gene inactivation was detected in nine of 58 (16%) tumors and was commonly associated with 17p losses. Tumors with 18q gains were significantly associated with a high number of chromosomal imbalances, primary nodal presentation, high serum lactate dehydrogenase levels, high International Prognostic Index, shorter cause-specific survival, and a high risk of relapse. Losses of 17p and p53 gene alterations were associated with an absence of complete response achievement. CONCLUSION: These results suggest that DLBCLs have a characteristic pattern of genomic alterations; 18q gains or amplifications and 17p losses are associated with particular clinicopathological features and aggressive clinical behavior. Additional studies are needed to confirm these observations in larger series of patients. PMID- 15337799 TI - Randomized, placebo-controlled study of oregovomab for consolidation of clinical remission in patients with advanced ovarian cancer. AB - PURPOSE: To assess oregovomab as consolidation treatment of advanced ovarian cancer and refine the immunotherapeutic strategy for subsequent study. PATIENTS AND METHODS: Patients with stage III/IV ovarian cancer who had a complete clinical response to primary treatment were randomly assigned to oregovomab or placebo administered at weeks 0, 4, and 8, and every 12 weeks up to 2 years or until recurrence. The primary end-point was time to relapse (TTR). RESULTS: One hundred forty-five patients were treated with oregovomab (n = 73) or placebo (n = 72). For the population overall, median TTR was not different between treatments at 13.3 months for oregovomab and 10.3 months for placebo (P =.71). Immune responses were induced in most actively treated patients. This was associated with prolonged TTR. Quality of life was not adversely impacted by treatment. Adverse events were reported with similar frequency in oregovomab and placebo groups, indicating a benign safety profile. A long-term survival follow-up is ongoing. Cox analysis of relapse data identified significant factors: performance status, CA-125 before third cycle, and baseline CA-125. Further evaluation identified a subpopulation with favorable prognostic indicators designated as the successful front-line therapy (SFLT) population. For the SFLT population, TTR was 24.0 months in the oregovomab group compared with 10.8 months for placebo (unadjusted hazard ratio of 0.543 [95% CI, 0.287 to 1.025]), a hypothesis generating observation. CONCLUSION: Consolidation therapy with oregovomab did not significantly improve TTR overall. A set of confirmatory phase III studies has been initiated to determine whether the SFLT population derives benefit from oregovomab treatment. PMID- 15337800 TI - The activity of taxanes in the treatment of sex cord-stromal ovarian tumors. AB - PURPOSE: To determine the efficacy and side effects of taxanes, with or without platinum, for the treatment of sex cord-stromal tumors of the ovary. PATIENTS AND METHODS: We conducted a retrospective review of all patients seen from 1985 to 2002 at The University of Texas M.D. Anderson Cancer Center with ovarian sex cord stromal tumors. Eligible patients underwent pathology confirmation and clinical evaluation at M.D. Anderson and received a taxane for initial or recurrent disease. RESULTS: Of 222 patients identified, 44 were eligible for analysis. For nine patients treated in the first-line adjuvant setting, median progression-free survival (PFS) was not reached at 51 months. Of two patients treated for measurable disease in the first-line setting, one had a complete response. Median PFS was 34.3 months; median overall survival (OS) was not reached. Median follow up was 90.3 months (range, 39.4 to 140.5 months). Response rate for 30 patients treated with a taxane +/- platinum for recurrent, measurable disease was 42%. Median PFS was 19.6 months; median OS was not reached. Median follow-up was 100.7 months (range, 8.1 to 361.3 months). The presence of platinum correlated with response in the recurrent, measurable disease setting. The number of patients was insufficient to detect relative efficacy of paclitaxel and docetaxel. Adverse effects of paclitaxel included neutropenia (n = 6), anemia (n = 1), thrombocytopenia (n = 1), myelodysplasia (n = 1), and hypersensitivity (n = 1). CONCLUSION: Taxanes seem to be active agents in the treatment of patients with sex cord-stromal tumors of the ovary. The combination of taxanes with platinum in the treatment of this disease deserves additional investigation. PMID- 15337801 TI - Estimating the cost of cancer: results on the basis of claims data analyses for cancer patients diagnosed with seven types of cancer during 1999 to 2000. AB - PURPOSE: Cancer accounts for 60.9 billion dollars in direct medical costs and 15.5 billion dollars for indirect morbidity costs. These estimates are derived primarily from national surveys or Federal databases. We derive estimates of the costs of cancer using administrative databases, which include claims and employment-related information on individuals insured by private or Medicare supplemental health plans. METHODS: A retrospective matched-cohort control analysis was performed using 1998 to 2000 databases with information on insurance claims, benefits, and health productivity for 3 million privately insured employees, their dependents, and early retirees. Study patients had new diagnoses of one of seven types of cancer (n = 12,709). Controls without cancer were matched at a 3:1 ratio by demographics. A variable follow-up length was used (maximum of 2 years). Direct costs included health care costs for patients and deductibles and copayments for caregivers. Indirect costs of work absence and short-term disability (STD) were calculated for a subgroup of cancer patients and caregivers. RESULTS: Mean monthly health care costs ranged from 2,187 dollars for prostate cancer to 7,616 dollars for pancreatic cancer, most often driven by hospitalization. Costs for controls were 329 dollars per month. Indirect morbidity costs to employees with cancer averaged 945 dollars, a result of a mean monthly loss of 2.0 workdays and 5.0 STD days. CONCLUSION: The economic burden of cancer is substantial. It is feasible to derive tumor-specific estimates of direct and indirect costs for large numbers of cancer patients using administrative databases. Policy makers charged with providing annual cost-of cancer estimates should incorporate data obtained from a broad range of sources. PMID- 15337802 TI - Papillary and follicular thyroid carcinomas show distinctly different microarray expression profiles and can be distinguished by a minimum of five genes. AB - PURPOSE: We have previously conducted independent microarray expression analyses of the two most common types of nonmedullary thyroid carcinoma, namely papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC). In this study, we sought to combine our data sets to shed light on the similarities and differences between these tumor types. MATERIALS AND METHODS: Microarray data from six PTCs, nine FTCs, and 13 normal thyroid samples were normalized to remove interlaboratory variability and then analyzed by unsupervised clustering, t test, and by comparison of absolute and change calls. Expression changes in four genes not previously implicated in thyroid carcinogenesis were verified by reverse transcriptase polymerase chain reaction on these same samples, together with eight additional FTC tumors. RESULTS: PTCs showed two distinct groups of genes that were either over- or underexpressed compared with normal thyroid, whereas the predominant changes in FTCs were of decreased expression. Five genes could collectively distinguish the two tumor types. PTCs showed overexpression of CITED1, claudin-10 (CLDN10), and insulin-like growth factor binding protein 6 (IGFBP6) but showed no change in expression of caveolin-1 (CAV1) or -2 (CAV2); conversely, FTCs did not express CLDN10 and had decreased expression of IGFBP6 and/or CAV1 and CAV2. CONCLUSION: PTC and FTC show distinctive microarray expression profiles, suggesting that either they have different molecular origins or they diverge distinctly from a common origin. Furthermore, if verified in a larger series of tumors, these genes could, in combination with known tumor specific chromosome translocations, form the basis of a valuable diagnostic tool. PMID- 15337803 TI - Radiation therapy with or without concurrent low-dose daily chemotherapy in locally advanced, nonmetastatic squamous cell carcinoma of the head and neck. AB - PURPOSE: To retrospectively investigate the difference between conventionally fractionated (CF) and hyperfractionated (Hfx) radiation therapy (RT), with and without either daily cisplatin (CDDP) or carboplatin (CBDCA), in patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN) enrolled onto two consecutive prospective randomized studies. PATIENTS AND METHODS: Treatment consisted of CF RT (70 Gy, group 1), CF RT and either daily CDDP (6 mg/m2) or daily CBDCA (25 mg/m2; group 2), Hfx RT (77 Gy, 1.1 Gy bid; group 3), or Hfx RT and daily CDDP (group 4). RESULTS: Hfx RT plus CDDP achieved better overall survival (OS) and local recurrence-free survival (LRFS) than any other group. There was an insignificant difference favoring Hfx RT over CF RT, either alone or in combination with CDDP or CBDCA, regarding both OS (P =.058 and P =.051, respectively) and LRFS (P =.088 and P =.091, respectively). No difference was seen between CF RT plus chemotherapy (CHT) and Hfx RT alone regarding either OS (P =.32) or LRFS (P =.48). Regional recurrence-free survival was similar in the four treatment groups. CF RT plus CHT and Hfx RT plus CDDP achieved better distant metastasis-free survival than CF RT and Hfx RT. High-grade toxicity was significantly more frequent in Hfx RT plus CDDP than in any other group, except in the Hfx RT group. Hfx RT led to significantly more acute toxicity and xerostomia than CF RT plus CHT. Hfx RT was more toxic than CF RT, either alone or with concurrent CHT. CONCLUSION: Results of this study show that there may be a therapeutic benefit for CF RT plus CHT over Hfx RT plus CDDP in patients with SCCHN, but this cannot be firmly established without a larger and well-planned controlled trial. PMID- 15337804 TI - Consolidation treatment with chimeric anti-GD2-antibody ch14.18 in children older than 1 year with metastatic neuroblastoma. AB - PURPOSE: Antibody treatment is considered tolerable and potentially effective in the therapy of neuroblastoma. We have analyzed stage 4 neuroblastoma patients older than 1 year who underwent consolidation treatment with the chimeric monoclonal anti-GD2-antibody ch14.18. PATIENTS AND METHODS: Stage 4 patients older than 1 year who completed initial treatment without event were eligible. ch14.18 was scheduled in a dose of 20 mg/m2/d during 5 days in six cycles every 2 months. Patients who did not receive ch14.18 served as controls. RESULTS: Of 334 assessable patients, 166 received ch14.18, 99 received a 12-month low-dose maintenance chemotherapy (MT) instead, and 69 had no additional treatment. During 695 ch14.18 cycles, fever (55% of cycles), abnormal C-reactive protein without infection (35%), cough (24%), rash (22%), and pain (16%) were the main side effects. Univariate analysis found similar event-free survival (EFS) for the three groups (3-year EFS, 46.5% +/- 4.1%, 44.4% +/- 4.9%, 37.1% +/- 5.9% for patients treated with antibody ch14.18, MT, and no additional therapy, respectively; log-rank test, P =.314). For overall survival (OS), ch14.18 treatment (3-year OS, 68.5% +/- 3.9%) was superior to MT (3-year OS, 56.6% +/- 5.0%) or no additional therapy (3-year OS, 46.8% +/- 6.2%; log-rank test, P =.018). Separate univariate analysis of patients with autologous stem-cell transplantation revealed no difference between patients with ch14.18 treatment and no additional consolidation. Multivariate analysis failed to demonstrate an advantage of antibody treatment for EFS and OS. CONCLUSION: Consolidation treatment of stage 4 neuroblastoma with ch14.18 was associated with considerable but manageable side effects. Compared with oral maintenance chemotherapy and no consolidation treatment, ch14.18 had no clear impact on the outcome of patients. PMID- 15337805 TI - Genetic variation in the leptin receptor gene and obesity in survivors of childhood acute lymphoblastic leukemia: a report from the Childhood Cancer Survivor Study. AB - PURPOSE: Overweight (body mass index [BMI] 25 to 29 kg/m2) and obesity (BMI > or = 30 kg/m2) frequently follow treatment for childhood acute lymphoblastic leukemia (ALL). Recent studies suggest that risk is most apparent in females treated with cranial radiation at a younger age. Because radiation at a young age may affect the hypothalamus causing leptin receptor insensitivity, we hypothesized that a polymorphism in the leptin receptor (LEPR) gene, Gln223Arg, might influence susceptibility to obesity in survivors of childhood ALL. PATIENTS AND METHODS: We genotyped 600 non-Hispanic white adult ALL survivors enrolled onto the Childhood Cancer Survivor Study. BMI was compared between those with two copies of the Arg allele to those who had at least one copy of the Gln allele. RESULTS: Female survivors with BMI > or = 25 kg/m2 were more likely Arg homozygous than those with BMI less than 25 kg/m2 (24% v 12%; P =.007). This difference was not observed in males. Moreover, among females treated with > or = 20 Gy cranial radiation, Arg/Arg individuals had six times higher odds of having BMI > or = 25 kg/m2 (95% CI, 2.1 to 22.0) than those with a Gln allele (P =.04 for interaction). CONCLUSION LEPR polymorphism may influence obesity in female survivors of childhood ALL, particularly those exposed to cranial radiation. Because obesity is associated with increased morbidity and mortality in later life, identification of children at high risk might allow for early targeted interventions. PMID- 15337806 TI - Treatment of children and adolescents with stage II testicular and stages I and II ovarian malignant germ cell tumors: A Pediatric Intergroup Study--Pediatric Oncology Group 9048 and Children's Cancer Group 8891. AB - PURPOSE: To determine whether children with localized gonadal malignant germ cell tumors (MGCT) stage II testicular and stages I and II ovarian treated with four cycles of standard-dose cisplatin combined with etoposide and low-dose bleomycin (PEB) have an event-free survival (EFS) of at least 85% without significant toxicity. PATIENTS AND METHODS: Between May 1990 and July 1995, eligible pediatric patients with stage II or recurrent from stage I (as a stage II) testicular MGCT and stages I and II ovarian MGCT were enrolled onto this Pediatric Oncology Group and Children's Cancer Group study. PEB chemotherapy consisted of bleomycin 15 U/m2 on day 1, cisplatin 20 mg/m2/d on days 1 to 5, and etoposide 100 mg/m2/d on days 1 to 5. Patients received four cycles of therapy at 21-day intervals. RESULTS: Seventy-four patients with a median age of 10.5 years (range, 8.7 months to 16.7 years) were enrolled. Primary sites included: stage II testicular (n = 17), stage I ovarian (n = 41), and stage II ovarian MGCT (n = 16). Treatment with standard PEB resulted in 6-year EFS of 95% and overall survival (OS) of 95.7%. EFS and OS by primary site were as follows: stage II testicular, 100% and 100%; stage I ovarian, 95.1% and 95.1%; and stage II ovarian, 87.5% and 93.8%, respectively. Two patients died from recurrent disease, and one patient died of secondary acute myelocytic leukemia. Infrequent grade 3 to 4 hematologic toxicity was reported. No grade 3 to 4 renal, pulmonary, or ototoxicity was observed. CONCLUSION: Combination chemotherapy with PEB results in excellent EFS and OS with minimal toxicity in children and adolescents with localized gonadal MGCT. PMID- 15337807 TI - Psychological, behavioral, and immune changes after a psychological intervention: a clinical trial. AB - PURPOSE: This randomized clinical trial tests the hypothesis that a psychological intervention can reduce emotional distress, improve health behaviors and dose intensity, and enhance immune responses. PATIENTS AND METHODS: We studied 227 women who were surgically treated for regional breast cancer. Before adjuvant therapy, women completed interviews and questionnaires assessing emotional distress, social adjustment, and health behaviors. A 60-mL blood sample was drawn for immune assays. Patients were randomly assigned to either the intervention group or assessment only group. The intervention was conducted in small patient groups, with one session per week for 4 months. The sessions included strategies to reduce stress, improve mood, alter health behaviors, and maintain adherence to cancer treatment and care. Reassessment occurred after completion of the intervention. RESULTS: As predicted, patients receiving the intervention showed significant lowering of anxiety, improvements in perceived social support, improved dietary habits, and reduction in smoking (all P <.05). Analyses of adjuvant chemotherapy dose-intensity revealed significantly more variability (ie, more dispersion in the dose-intensity values) for the assessment arm (P <.05). Immune responses for the intervention patients paralleled their psychological and behavioral improvements. T-cell proliferation in response to phytohemagglutinin and concanavalin A remained stable or increased for the Intervention patients, whereas both responses declined for Assessment patients; this effect was replicated across three concentrations for each assay (all P <.01). CONCLUSION: These data show a convergence of significant psychological, health behavior, and biologic effects after a psychological intervention for cancer patients. PMID- 15337808 TI - Survey on use of palliative radiotherapy in hospice care. AB - PURPOSE: Radiation oncologists and hospice professionals both provide end-of-life care for oncology patients, and little has been written about the interface between these two groups of specialists. Hospice professionals were surveyed to assess the perceived need for palliative radiotherapy in the hospice setting, to investigate factors that limit the access of hospice patients to radiotherapy, and to suggest areas of future collaboration on education, research, and patient advocacy. PATIENTS AND METHODS: Members of the National Hospice and Palliative Care Organization (NHPCO) and American Society for Therapeutic Radiology and Oncology jointly authored a questionnaire to investigate the beliefs of hospice professionals toward the use of radiotherapy for oncology patients in hospice. The questionnaire was distributed to all NHPCO member institutions, and the results were compiled and statistically analyzed. RESULTS: Four hundred eighty of more than 1,800 surveyed facilities responded to the questionnaire. The findings suggest that the majority of hospice professionals feel that radiotherapy is important in palliative oncology and that radiotherapy is widely available in the United States. Yet less than 3% on average of hospice patients served by hospices responding to the survey actually received radiotherapy in 2002. The most common barriers to radiotherapy in hospice care include radiotherapy expense, transportation difficulties, short life expectancy, and educational deficiencies between the specialties. CONCLUSION: Multiple barriers act to limit the use of palliative radiotherapy in hospice care. Finding ways to surmount these obstacles will provide opportunity for improvement in the end-of-life care of cancer patients. PMID- 15337809 TI - Efficacy and safety of ibandronate in the treatment of opioid-resistant bone pain associated with metastatic bone disease: a pilot study. AB - PURPOSE: Bone metastases are associated with severe and sometimes intractable pain, compromising patient quality of life (QOL). This open-label pilot study investigated the effects of short-term intensive treatment with intravenous (i.v.) ibandronate on opioid-resistant bone pain in patients with skeletal metastases. PATIENTS AND METHODS: Eighteen patients with advanced tumors and metastatic bone disease received nonstandard treatment with 4 mg of ibandronate administered i.v. (2-hour infusion) for 4 consecutive days (16-mg total dose). Baseline opioid analgesic use was equivalent to 400 mg/d of morphine. Patients were assessed for 6 weeks or until death. Changes from baseline were determined for bone pain, opioid consumption, patient functioning, QOL, performance status, and biochemical markers of calcium metabolism and bone turnover. Renal function was assessed by serum urea and creatinine measurement. RESULTS: Short-term, intensive ibandronate treatment significantly reduced bone pain scores within 7 days (P <.001). Pain reductions were sustained over the study period. Ibandronate significantly improved QOL, patient functioning, and performance status (P <.05). Mean values of the urinary cross-links pyridinoline and deoxypyridinoline tended to increase after day 21, returning close to baseline values by day 42. There was no correlation between the change in crosslinks values and the change in pain scores after ibandronate treatment. Ibandronate was well tolerated, with no evidence of renal toxicity. CONCLUSION: Nonstandard, intensive treatment with i.v. ibandronate seems to have a marked analgesic effect in patients with opioid resistant bone pain from metastatic bone disease. Further investigation is warranted. PMID- 15337810 TI - Receptor imaging in oncology by means of nuclear medicine: current status. AB - To date, our understanding of the role of receptors and their cognate ligands in cancer is being successfully translated into the design and development of an arsenal of new, less toxic, and more specific anticancer drugs. Because most of these novel drugs are cytostatic, objective response as measured by morphologic imaging modalities (eg, computed tomography or magnetic resonance imaging) cannot be used as a surrogate marker for drug development or for clinical decision making. Positron emission tomography (PET) can be used to image and quantify the in vivo distribution of positron-emitting radioisotopes such as oxygen-15, carbon 11, and fluorine-18 that can be substituted or added into biologically relevant and specific receptor radioligands. Similarly, single-photon emission computed tomography (SPECT) can be used to image and quantify the in vivo distribution of receptor targeting compounds labeled with indium-111, technetium-99m, and iodine 123. By virtue of their whole-body imaging capacity and the absence of errors of sampling and tissue manipulation as well as preparation, both techniques have the potential to address locoregional receptor status noninvasively and repetitively. This article reviews available data on the in vivo evaluation of receptor systems by means of PET or SPECT for identifying and monitoring patients with sufficient receptor overexpression for tailored therapeutic interventions, and also for depicting tumor tissue and determining the currently largely unknown heterogeneity in receptor expression among different tumor lesions within and between patients. PMID- 15337811 TI - CNS metastases in breast cancer. AB - As systemic therapy of metastatic breast cancer improves, CNS involvement is becoming a more widespread problem. This article summarizes the current knowledge regarding the incidence, clinical presentation, diagnosis, prognosis, and treatment of CNS metastases in patients with breast cancer. When available, studies specific to breast cancer are presented; in studies in which many solid tumors were evaluated together, the proportion of patients with breast cancer is noted. On the basis of data from randomized trials and retrospective series, neurosurgery and stereotactic radiosurgery (SRS) may prolong survival in patients with single brain metastases. The treatment of multiple metastases remains controversial, as does the routine use of whole-brain radiotherapy (WBRT) after either surgery or SRS. Although it is widely assumed that chemotherapy is of limited benefit, data from case series and case reports suggest otherwise. WBRT, neurosurgery, SRS, and medical therapy each have a role in the treatment of CNS metastases; however, neurologic symptoms frequently are not fully reversible, even with appropriate therapy. Studies specifically targeted toward this group of patients are needed. PMID- 15337812 TI - Challenging manifestations of malignancies. Case 1. Polycythemia and high serum erythropoietin level as a result of hemangioblastoma. PMID- 15337813 TI - Challenging manifestations of malignancies. Case 2. Langerhans cell histiocytosis presenting with a skin rash. PMID- 15337814 TI - Challenging manifestations of malignancies. Case 3. Fibrinous tracheitis: an uncommon manifestation of Hodgkin's lymphoma. PMID- 15337815 TI - Pulling the trigger. PMID- 15337816 TI - Testing for epidermal growth factor receptor in lung cancer: have we learned anything from HER-2 testing? PMID- 15337818 TI - Ion channel structure and the promise of bacteria: cyclic nucleotide-gated channels in the queue. PMID- 15337819 TI - A cyclic nucleotide modulated prokaryotic K+ channel. AB - A search of prokaryotic genomes uncovered a gene from Mesorhizobium loti homologous to eukaryotic K(+) channels of the S4 superfamily that also carry a cyclic nucleotide binding domain at the COOH terminus. The gene was cloned from genomic DNA, and the protein, denoted MloK1, was overexpressed in Escherichia coli and purified. Gel filtration analysis revealed a heterogeneous distribution of protein sizes which, upon inclusion of cyclic nucleotide, coalesces into a homogeneous population, eluting at the size expected for a homotetramer. As followed by a radioactive (86)Rb(+) flux assay, the putative channel protein catalyzes ionic flux with a selectivity expected for a K(+) channel. Ion transport is stimulated by cAMP and cGMP at submicromolar concentrations. Since this bacterial homologue does not have the "C-linker" sequence found in all eukaryotic S4-type cyclic nucleotide-modulated ion channels, these results show that this four-helix structure is not a general requirement for transducing the cyclic nucleotide-binding signal to channel opening. PMID- 15337820 TI - How source content determines intracellular Ca2+ release kinetics. Simultaneous measurement of [Ca2+] transients and [H+] displacement in skeletal muscle. AB - In skeletal muscle, the waveform of Ca(2+) release under clamp depolarization exhibits an early peak. Its decay reflects an inactivation, which locally corresponds to the termination of Ca(2+) sparks, and is crucial for rapid control. In cardiac muscle, both the frequency of spontaneous sparks (i.e., their activation) and their termination appear to be strongly dependent on the Ca(2+) content in the sarcoplasmic reticulum (SR). In skeletal muscle, no such role is established. Seeking a robust measurement of Ca(2+) release and a way to reliably modify the SR content, we combined in the same cells the "EGTA/phenol red" method (Pape et al., 1995) to evaluate Ca(2+) release, with the "removal" method (Melzer et al., 1987) to evaluate release flux. The cytosol of voltage-clamped frog fibers was equilibrated with EGTA (36 mM), antipyrylazo III, and phenol red, and absorbance changes were monitored simultaneously at three wavelengths, affording largely independent evaluations of Delta[H(+)] and Delta[Ca(2+)] from which the amount of released Ca(2+) and the release flux were independently derived. Both methods yielded mutually consistent evaluations of flux. While the removal method gave a better kinetic picture of the release waveform, EGTA/phenol red provided continuous reproducible measures of calcium in the SR (Ca(SR)). Steady release permeability (P), reached at the end of a 120-ms pulse, increased as Ca(SR) was progressively reduced by a prior conditioning pulse, reaching 2.34-fold at 25% of resting Ca(SR) (four cells). Peak P, reached early during a pulse, increased proportionally much less with SR depletion, decreasing at very low Ca(SR). The increase in steady P upon depletion was associated with a slowing of the rate of decay of P after the peak (i.e., a slower inactivation of Ca(2+) release). These results are consistent with a major inhibitory effect of cytosolic (rather than intra-SR) Ca(2+) on the activity of Ca(2+) release channels. PMID- 15337821 TI - Using total fluorescence increase (signal mass) to determine the Ca2+ current underlying localized Ca2+ events. AB - The feasibility of determining localized Ca(2+) influx using only wide-field fluorescence images was explored by imaging (using fluo-3) single channel Ca(2+) fluorescence transients (SCCaFTs), due to Ca(2+) entry through single openings of Ca(2+)-permeable ion channels, while recording unitary channel currents. Since the image obtained with wide-field optics is an integration of both in-focus and out-of-focus light, the total fluorescence increase (DeltaF(total) or "signal mass") associated with a SCCaFT can be measured directly from the image by adding together the fluorescence increase due to Ca(2+) influx in all of the pixels. The assumptions necessary for obtaining the signal mass from confocal linescan images are not required. Two- and three-dimensional imaging was used to show that DeltaF(total) is essentially independent of the position of the channel with respect to the focal plane of the microscope. The relationship between Ca(2+) influx and DeltaF(total) was obtained using SCCaFTs from plasma membrane caffeine activated cation channels when Ca(2+) was the only charge carrier of the inward current. This relationship was found to be linear, with the value of the slope (or converting factor) affected by the particular imaging system set-up, the experimental conditions, and the properties of the fluorescent indicator, including its binding capacity with respect to other cellular buffers. The converting factor was used to estimate the Ca(2+) current passing through caffeine-activated channels in near physiological saline and to estimate the endogenous buffer binding capacity. In addition, it allowed a more accurate estimate of the Ca(2+) current underlying Ca(2+) sparks resulting from Ca(2+) release from intracellular stores via ryanodine receptors in the same preparation. PMID- 15337822 TI - Angiotensin II (AT1) receptors and NADPH oxidase regulate Cl- current elicited by beta1 integrin stretch in rabbit ventricular myocytes. AB - Direct stretch of beta1 integrin activates an outwardly rectifying, tamoxifen sensitive Cl(-) current (Cl(-) SAC) via focal adhesion kinase (FAK) and/or Src. The characteristics of Cl(-) SAC resemble those of the volume-sensitive Cl(-) current, I(Cl,swell). Because myocyte stretch releases angiotensin II (AngII), which binds AT1 receptors (AT1R) and stimulates FAK and Src in an autocrine paracrine loop, we tested whether AT1R and their downstream signaling cascade participate in mechanotransduction. Paramagnetic beads coated with mAb for beta1 integrin were applied to myocytes and pulled upward with an electromagnet while recording whole-cell anion current. Losartan (5 microM), an AT1R competitive antagonist, blocked Cl(-) SAC but did not significantly alter the background Cl( ) current in the absence of integrin stretch. AT1R signaling is mediated largely by H(2)O(2) produced from superoxide generated by sarcolemmal NADPH oxidase. Diphenyleneiodonium (DPI, 60 microM), a potent NADPH oxidase inhibitor, rapidly and completely blocked both Cl(-) SAC elicited by stretch and the background Cl( ) current. A structurally unrelated NADPH oxidase inhibitor, 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF, 0.5 and 2 mM), also rapidly and completely blocked Cl(-) SAC as well as a large fraction of the background Cl(-) current. With continuing integrin stretch, Cl(-) SAC recovered upon washout of AEBSF (2 mM). In the absence of stretch, exogenous AngII (5 nM) activated an outwardly rectifying Cl(-) current that was rapidly and completely blocked by DPI (60 microM). Moreover, exogenous H(2)O(2) (10, 100, and 500 microM), the eventual product of NADPH oxidase activity, also activated Cl(-) SAC in the absence of stretch, whereas catalase (1,000 U/ml), an H(2)O(2) scavenger, attenuated the response to stretch. Application of H(2)O(2) during NADPH oxidase inhibition by either DPI (60 microM) or AEBSF (0.5 mM) did not fully reactivate Cl(-) SAC, however. These results suggest that stretch of beta1-integrin in cardiac myocytes elicits Cl(-) SAC by activating AT1R and NADPH oxidase and, thereby, producing reactive oxygen species. In addition, NADPH oxidase may be intimately coupled to the channel responsible for Cl(-) SAC, providing a second regulatory pathway. PMID- 15337823 TI - Complementary and alternative medicine: opportunities for education and research. PMID- 15337824 TI - Citrus aurantium, an ingredient of dietary supplements marketed for weight loss: current status of clinical and basic research. AB - Seville orange (Citrus aurantium) extracts are being marketed as a safe alternative to ephedra in herbal weight-loss products, but C. aurantium may also have the potential to cause adverse health effects. C. aurantium contains synephrine (oxedrine), which is structurally similar to epinephrine. Although no adverse events have been associated with ingestion of C. aurantium products thus far, synephrine increases blood pressure in humans and other species, and has the potential to increase cardiovascular events. Additionally, C. aurantium contains 6',7'-dihydroxybergamottin and bergapten, both of which inhibit cytochrome P450 3A, and would be expected to increase serum levels of many drugs. There is little evidence that products containing C. aurantium are an effective aid to weight loss. Synephrine has lipolytic effects in human fat cells only at high doses, and octopamine does not have lipolytic effects in human adipocytes. PMID- 15337825 TI - Genomic approaches that aid in the identification of transcription factor target genes. AB - It is well-established that deregulation of the transcriptional activity of many different genes has been causatively linked to human diseases. In cancer, altered patterns of gene expression are often the result of the inappropriate expression of a specific transcriptional activator or repressor. Functional studies of cancer-specific transcription factors have relied upon the study of candidate target genes. More recently, gene expression profiling using DNA microarrays that contain tens of thousands of cDNAs corresponding to human mRNAs has allowed for a large-scale identification of genes that respond to increased or decreased levels of a particular transcription factor. However, such experiments do not distinguish direct versus indirect target genes. Coupling chromatin immunoprecipitation to micro-arrays that contain genomic regions (ChIP-chip) has provided investigators with the ability to identify, in a high-throughput manner, promoters directly bound by specific transcription factors. Clearly, knowledge gained from both types of arrays provides complementary information, allowing greater confidence that a transcription factor regulates a particular gene. In this review, we focus on Polycomb group (PcG) complexes as an example of transcriptional regulators that are implicated in various cellular processes but about which very little is known concerning their target gene specificity. We provide examples of how both expression arrays and ChIP-chip microarray-based assays can be used to identify target genes of a particular PcG complex and suggest improvements in the application of array technology for faster and more comprehensive identification of directly regulated target genes. PMID- 15337826 TI - The consequences of exhaustive antiestrogen therapy in breast cancer: estrogen induced tumor cell death. AB - Forty years ago, the endocrine treatment for breast cancer was a last resort at palliation before the disease overwhelmed the patient (1). Ovarian ablation was the treatment of choice for the premenopausal patient, whereas either adrenalectomy or, paradoxically, high-dose synthetic estrogen therapy were used for treatment in postmenopausal patients. A reduction or an excess of estrogen provoked objective responses in one out of three women. Unfortunately, there was no way of predicting who would respond to endocrine ablation, and because so few patients responded there was no enthusiasm for developing new endocrine agents. All hopes for a cure for breast cancer turned to appropriate combinations of cytotoxic chemotherapy. Today tamoxifen, a nonsteroidal antiestrogen (2), has proven to be effective in all stages of premenopausal and postmenopausal breast cancer, and several new endocrine strategies, including aromatase inhibitors, luteinizing-hormone releasing hormone (LHRH) superagonists, and a pure antiestrogen (fulvestrant), are now available for breast cancer treatment. Additionally, tamoxifen and raloxifene, a related compound, are used to reduce the risk of breast cancer and osteoporosis, respectively, in high-risk groups (3). Hormonal modulation and strategies to prevent the actions of estrogen in the breast are ubiquitous. However, with successful changes in treatment strategies comes the consequence of change. This minireview will describe the current strategies for the treatment and prevention of breast cancer and present emerging new concepts about the consequences of exhaustive antiestrogen treatment on therapeutic resistance. PMID- 15337827 TI - Structures of integrin domains and concerted conformational changes in the bidirectional signaling mechanism of alphaIIbbeta3. AB - Integrins are heterodimeric type I transmembrane cell-adhesive receptors whose affinity for ligands is regulated by tertiary and quaternary conformational changes that are transmitted from the cytoplasmic tails to the extracellular ectodomains during the transition from the inactive to the active state. Receptor occupancy initiates further structural alterations that transduce signals across the plasma membrane and result in receptor clustering and recruitment of signaling molecules and cytoskeletal rearrangements at the integrin's cytoplasmic domains. The large distance between the intracellular cytoplasmic domains and the ligand-binding site, which in an extended conformation spans more that 200 A, imposes a complex mechanism of interdomain communication for the bidirectional information flow across the plasma membrane. Significant progress has recently been made in elucidating the crystal and electron microscopy structures of integrin ectodomains in its unliganded and liganded states, and the nuclear magnetic resonance solution structures of stalk domains and the cytoplasmic tails. These structures revealed the location of sites that are functionally important and provided the basis for defining new models of integrin activation and signaling through bidirectional conformational changes, and for understanding the structural basis of the cation-dependent ligand-binding specificity of integrins. Platelet integrin alphaIIbbeta3 has served as a paradigm for many aspects of the structure and function of integrins The aim of this minireview is to combine recent structural and biochemical studies on integrin receptors that converge into a model of the tertiary and quaternary conformational changes in alphaIIbbeta3 and other homologous integrins that propagate inside-out and outside-in signals. PMID- 15337828 TI - Tocotrienol-induced caspase-8 activation is unrelated to death receptor apoptotic signaling in neoplastic mammary epithelial cells. AB - Tocotrienols, a subclass in the vitamin E family of compounds, have been shown to induce apoptosis by activating caspase-8 and caspase-3 in neoplastic mammary epithelial cells. Since caspase-8 activation is associated with death receptor apoptotic signaling, studies were conducted to determine the exact death receptor/ligand involved in tocotrienol-induced apoptosis. Highly malignant +SA mouse mammary epithelial cells were grown in culture and maintained in serum-free media. Treatment with 20 microM gamma-tocotrienol decreased+SA cell viability by inducing apoptosis, as determined by positive terminal dUTP nick end labeling (TUNEL) immunocytochemical staining. Western blot analysis showed that gamma tocotrienol treatment increased the levels of cleaved (active) caspase-8 and caspase-3. Combined treatment with caspase inhibitors completely blocked tocotrienol-induced apoptosis. Additional studies showed that treatment with 100 ng/ml tumor necrosis factor-alpha (TNF-alpha), 100 ng/ml FasL, 100 ng/ml TNF related apoptosis-inducing ligand (TRAIL), or 1 microg/ml apoptosis-inducing Fas antibody failed to induce death in +SA cells, indicating that this mammary tumor cell line is resistant to death receptor-induced apoptosis. Furthermore, treatment with 20 microM gamma-tocotrienol had no effect on total, membrane, or cytosolic levels of Fas, Fas ligand (FasL), or Fas-associated via death domain (FADD) and did not induce translocation of Fas, FasL, or FADD from the cytosolic to the membrane fraction, providing additional evidence that tocotrienol-induced caspase-8 activation is not associated with death receptor apoptotic signaling. Other studies showed that treatment with 20 microM gamma-tocotrienol induced a large decrease in the relative intracellular levels of phospho phosphatidylinositol 3-kinase (PI3K)-dependent kinase 1 (phospho-PDK-1 active), phospho-Akt (active), and phospho-glycogen synthase kinase3, as well as decreasing intracellular levels of FLICE-inhibitory protein (FLIP), an antiapoptotic protein that inhibits caspase-8 activation, in these cells. Since stimulation of the PI3K/PDK/Akt mitogenic pathway is associated with increased FLIP expression, enhanced cellular proliferation, and survival, these results indicate that tocotrienol-induced caspase-8 activation and apoptosis in malignant +SA mammary epithelial cells is associated with a suppression in PI3K/PDK-1/Akt mitogenic signaling and subsequent reduction in intracellular FLIP levels. PMID- 15337829 TI - Cu,Zn-superoxide dismutase is lower and copper chaperone CCS is higher in erythrocytes of copper-deficient rats and mice. AB - Discovery of a sensitive blood biochemical marker of copper status would be valuable for assessing marginal copper intakes. Rodent models were used to investigate whether erythrocyte concentrations of copper,zinc-superoxide dismutase (SOD), and the copper metallochaperone for SOD (CCS) were sensitive to dietary copper changes. Several models of copper deficiency were studied in postweanling male Holtzman rats, male Swiss Webster mice offspring, and both rat and mouse dams. Treatment resulted in variable but significantly altered copper status as evaluated by the presence of anemia, and lower liver copper and higher liver iron concentrations in copper-deficient compared with copper-adequate animals. Associated with this copper deficiency were consistent reductions in immunoreactive SOD and robust enhancements in CCS. In most cases, the ratio of CCS:SOD was several-fold higher in red blood cell extracts from copper-deficient compared with copper-adequate rodents. Determination of red cell CCS:SOD may be useful for assessing copper status of humans. PMID- 15337830 TI - Cocoa procyanidins inhibit proliferation and angiogenic signals in human dermal microvascular endothelial cells following stimulation by low-level H2O2. AB - Procyanidins extracted from cocoa play a role in the defense against oxidative stress, as well as in vascular and immune functions. We previously reported that pentameric procyanidins isolated from cocoa inhibit the expression of the tyrosine kinase ErbB2 gene, thus slowing the growth of cultured human aortic endothelial cells. We herein investigate the further consequences of such inhibition by cocoa procyanidins, particularly regarding the protein level in phosphorylation patterns and the effects on the proliferation of human dermal microvascular endothelial cells (HDMECs) following angiogenic stimulation with low-level H2O2. We report herein that both the pentameric and octameric procyanidin fractions of cocoa inhibit the proliferation of HDMECs, whereas the pentameric fraction modulates the activity of several crucial proteins in angiogenic signaling by altering their tyrosine phosphorylation. Similar to aortic endothelial cells, the pentameric procyanidin fraction down-regulates the expression of ErbB2 tyrosine kinase in HDMECs. In conclusion, we report evidence suggesting that polyphenols may influence endothelial growth signaling, thus affecting angiogenesis in vitro. If these observations are applicable in vivo, they suggest a beneficial effect for cells overexpressing ErbB2, such as in specific neoplasias PMID- 15337831 TI - Differential increases in syntheses of newly identified trypsinogen 2 isoforms by dietary protein in rat pancreas. AB - We have found that dietary protein markedly induced pancreatic serine protease activity via a mechanism independent of luminal trypsin activity in pancreaticobiliary-diverted (PBD) rats. The aim of this study was to examine the effects of dietary protein on the synthesis of trypsinogen isoforms by comparing in vivo incorporation of [35S] L-methionine into isoform proteins in PBD and sham operated rats. A small duodenal segment including the ampulla of Vater was sectioned and transposed to the upper ileum with end-to-side anastomosis (PBD) or duodenal transection was followed by reanastomosis (sham) in male Sprague-Dawley rats. After recovery, PBD and sham rats were fed a 25% or 60% casein-sucrose based diet (NC or HC) for 14 days. Rats were then intravenously injected with [35S] L-methionine (15 MBq/kg body weight) 30 mins before being sacrificed for analysis of pancreatic enzymes by two-dimensional SDS-polyacrylamide gel electrophoresis. By using electrophoresis with narrow range of isoelectric focusing (pI 4.5-5.5), five trypsinogen 2 (2-x) isoform spots were identified using both [35S] incorporation and Coomassie brilliant blue (CBB) staining in PBD rats, but not in sham rats. N-terminal sequences of these trypsinogen 2-x spots were identical to known rat trypsinogen 2 with the exception that the third valine was changed to isoleucine in one isoform. In PBD rats, feeding of HC specifically increased the [35S] and CBB intensities of these trypsinogen 2-x isoforms and trypsinogen 3. The degree of induction of the five trypsinogen 2-x molecules by HC varied greatly. Trypsinogen 1 and 4, which are the major trypsinogens in normal rats, showed no changes. We conclude that increases in synthesis of a few newly identified trypsinogen 2-x isoforms mainly contribute to the induction of trypsin activity in the pancreas by HC in PBD rats. PMID- 15337832 TI - Differential translational effects of myristic acid and eicosapentaenoic acid on 3-hydroxy-3-methylglutaryl-CoA reductase from Reuber H35 hepatoma cells. AB - The mechanisms by which saturated and polyunsaturated fatty acids may exert their effects on levels of blood cholesterol and human atherosclerosis have not been fully established. In this work, we studied the translational effects of myristic (14:0) and eicosapentaenoic (20:5) acids on 3-hydroxy-3-methylglutaryl-CoA (HMG CoA) reductase from Reuber H35 hepatoma cells. This enzyme is an intrinsic membrane, 96-kDa protein whose proteolysis releases an enzymatically active, 52- to 56-kDa, soluble fragment. We optimized an immunoblot procedure for quantifying small amounts of both the native and the soluble forms of HMG-CoA reductase from Reuber H35 hepatoma cells. We demonstrated that the upregulation of HMG-CoA reductase by a acid is due to an increase of the HMG-CoA reductase protein; therefore, protein synthesis would be required for the increase of HMG-CoA reductase activity caused by this fatty acid. In contrast, the downregulation of HMG-CoA reductase caused by eicosapentaenoic acid is not due to decreased protein synthesis, since similar levels of protein were found in the presence and absence of this fatty acid. Results obtained with cycloheximide as a protein-synthesis inhibitor confirm these findings. PMID- 15337833 TI - Resistant starch fraction prepared from kintoki bean affects gene expression of genes associated with cholesterol metabolism in rats. AB - Feeding rats beans with resistant starch reduces the serum cholesterol concentration; however, the mechanism is not fully understood. We examined the effects of resistant starch of kintoki (Phaseolus vulgaris, variety) bean on serum cholesterol and hepatic mRNAs in rats. Male F344/Du Crj rats were fed a cholesterol-free diet either with 5 g of cellulose powder (control)/100 g or 5 g of pancreatin-resistant fraction prepared from kintoki bean (kintoki)/100 g diet for 4 weeks. There were no differences in the body weight gain, food intake, liver weight, and mass of cecum contents between the groups. Serum total cholesterol, very low density lipoprotein (VLDL) + intermediate density lipoprotein (IDL) + low density lipoprotein (LDL)-cholesterol, and high density lipoprotein (HDL)-cholesterol levels in the kintoki group were significantly (at least P < 0.05) lower than in the control group throughout the feeding period. There was no difference in the serum triglyceride concentration between two groups throughout the feeding period. Total hepatic cholesterol in the control group was significantly (P < 0.01) lower than in the kintoki groups. Fecal bile acid, cecal acetate, propionate and n-butyrate concentrations in the kintoki group all were significantly (P < 0.05) higher than in the control group. Likewise, hepatic cholesterol 7alpha-hydroxylase, LDL receptor, and SR-B1 mRNA levels in the kintoki group were significantly (P < 0.05) higher than in the control group. The results suggest that resistant starch of kintoki bean reduces serum cholesterol level by increasing hepatic LDL receptor, SR-B1, and cholesterol 7alpha-hydroxylase mRNAs. PMID- 15337834 TI - Involvement of the peripheral cholinergic muscarinic system in the compensatory ovarian hypertrophy in the rat. AB - In the present experiments, unilateral ovariectomy (ULO) induced compensatory hypertrophy (COH) of the remaining rat ovary (60%-85% increase in ovarian weight, total proteins, and total RNA and DNA). An increased thymidine uptake preceded the organ enlargement. COH was inhibited by i.p.-administered muscarinic antagonist propantheline (dose-dependently) or botulinum toxin delivered locally to the ovary. The effects were reversed by bethanecol i.p. (a muscarinic agonist). In sham ULO animals, [3H]-scopolamine binding to ovarian membranes indicated the existence of muscarinic receptors (Kd 2.5 nM, Bmax 12 fmol/mg proteins, Hill 1.0). The ovarian 1,2-diacylglycerol (DAG) was 120-150 pmol/mg tissue and did not react to carbachol in vitro (50 microM). At 15 minutes after ULO, the [3H]-scopolamine binding was unchanged (Kd 2.6 nM, Bmax 12.6 fmol/mg tissue, Hill 1.0), but the ovarian DAG was increased (280-350 pmol/mg tissue) and increased further in response to carbachol (460-550 pmol/mg tissue). After ULO, ovarian DAG remained continuously responsive to carbachol. The ULO-induced DAG increase and enhanced susceptibility to carbachol were inhibited by the botulinum toxin or atropine pretreatments. Abdominal vagotomy done immediately before ULO also inhibited the ULO-induced DAG increase and DAG responsiveness to carbachol. However, when the vagotomy was performed 10 mins after ULO, the ovarian DAG remained responsive to carbachol in vitro. The data suggest that the peripheral cholinergic system, including the ovarian muscarinic receptors, stimulates COH. This is apparently associated with the ULO-induced coupling of the ovarian muscarinic receptors to phosphoinositide (PI) breakdown. Vagus plays a role in the occurrence of the changed muscarinic receptor-PI breakdown relationship in the remaining ovary. PMID- 15337835 TI - Intermittent hypoxic training protects canine myocardium from infarction. AB - This investigation examined cardiac protective effects of normobaric intermittent hypoxia training. Six dogs underwent intermittent hypoxic training for 20 consecutive days in a normobaric chamber ventilated intermittently with N2 to reduce fraction of inspired oxygen (FiO2) to 9.5%-10%. Hypoxic periods, initially 5 mins and increasing to 10 mins, were followed by 4-min normoxic periods. This hypoxia-normoxia protocol was repeated, initially 5 times and increasing to 8 times. The dogs showed no discomfort during intermittent hypoxic training. After 20 days of hypoxic training, the resistance of ventricular myocardium to infarction was assessed in an acute experiment. The left anterior descending (LAD) coronary artery was occluded for 60 mins and then reperfused for 5 hrs. At 30 mins of LAD occlusion, radioactive microspheres were injected through a left atrial catheter to assess coronary collateral blood flow into the ischemic region. After 5 hrs reperfusion, the heart was dyed to delineate the area at risk (AAR) of infarction and stained with triphenyl tetrazolium chloride to identify infarcted myocardium. During LAD occlusion and reperfusion, systemic hemodynamics and global left ventricular function were stable. Infarction was not detected in 4 hearts and was 1.6% of AAR in the other 2 hearts. In contrast, 6 dogs sham trained in a chamber ventilated with compressed air and 5 untrained dogs subjected to the same LAD occlusion/reperfusion protocol had infarcts of 36.8% +/ 5.8% and 35.2% +/- 9.5% of the AAR, respectively. The reduction in infarct size of four of the six hypoxia-trained dogs could not be explained by enhanced collateral blood flow to the AAR. Hypoxia-trained dogs had no ventricular tachycardia or ventricular fibrillation. Three sham-trained dogs had ventricular tachycardia and two had ventricular fibrillation. Three untrained dogs had ventricular fibrillation. In conclusion, intermittent hypoxic training protects canine myocardium from infarction and life-threatening arrhythmias during coronary artery occlusion and reperfusion. The mechanism responsible for this potent cardioprotection merits further study. PMID- 15337836 TI - Effects of cardiac hormones on arterial pressure and sodium excretion in NPRA knockout mice. AB - These studies were designed to determine if the atria contains natriuretic substances that act through a non-natriuretic peptide type A (NPRA) receptor mechanism. C57BL/6 mice, either wild-type NPRA++ (WT) or NPRA-- knockout (KO), were anesthetized with pentobarbital. Catheters were placed in the trachea, carotid artery, jugular vein, and bladder. Urine was collected for six 30-min periods. Both groups received an iv injection of 100 ng of rat atrial natriuretic peptide (rANP) in 200 microl of saline after the first period (30 mins) and 200 microl of rat atrial extract after the fourth period (120 mins). ANP injection increased urine flow (UF) to 2.7 +/- 0.5 microl/min in the WT versus 1.9 +/- 0.2 in KO. Extract increased UF to 7.9 +/- 1.5 microl/min in WT versus 2.7 +/- 0.4 in KO (P < 0.01). ANP increased sodium excretion (ENa) to 0.47 +/- 0.10 micromoles/min in WT versus 0.27 +/- 0.04 in KO (P < 0.05). Extract increased ENa to 1.44 +/- 0.47 micromoles/min in WT versus 0.26 +/- 0.06 in KO (P < 0.05). Extract decreased mean arterial pressure (MAP) to 62 +/- 3 mm Hg in the WT versus 81 +/- 5 in KO (P < 0.01). ENa and MAP responses to extract in KO were not different from responses to 200 microl of saline. A constant 150-min infusion of rat atrial extract increased urine flow by 3-fold and ENa by 5-fold (both P < 0.05) in the WT mice but had no significant effect in the KO mice. Thus, acute renal and MAP responses to atrial extracts require the NPRA receptor. PMID- 15337837 TI - Chronic prenatal exposure to cocaine alters cerebrovascular responses in newborn pigs. AB - Maternal cocaine abuse may increase the incidence of perinatal asphyxia. In nonexposed asphyxiated neonates, decreased cerebrospinal fluid (CSF) cAMP concentrations are associated with poor neurological outcome. On the other hand, cocaine increases central nervous system (CNS) cAMP. Therefore, we hypothesized that in utero cocaine exposure may increase brain cAMP and thereby preserve cerebrovascular responses to cAMP-dependent stimuli following asphyxia. Pregnant pigs received either cocaine (1 mg/kg, i.v.) twice weekly during the last trimester or normal saline vehicle (sham-control) and were allowed to deliver vaginally at term. Cranial windows were implanted in the newborn pigs within the first week of life and used to collect CSF for cAMP determinations and to assess changes in pial arteriolar diameters (PAD). In the first part of the study, pial arteriolar responses to different vasodilator and vasoconstrictor stimuli were evaluated in piglets prior to asphyxia (n = 20). In newborn pigs exposed to cocaine, cerebrovascular responses to hypercapnia and norepinephrine were significantly exaggerated compared to controls. Then, piglets were randomly selected for the second part of the study that involved prolonged asphyxia (n = 12). In cocaine-exposed but not sham-control piglets, CSF cAMP increased markedly during asphyxia. In the sham piglets, but not the cocaine-exposed piglets, CSF cAMP fell progressively below the baseline during recovery. Cerebrovascular reactivity to cAMP-dependent stimuli (hypercapnia and isoproterenol) was preserved during recovery from asphyxia in the cocaine-exposed piglets but significantly attenuated in the sham controls. We conclude that piglets with chronic prenatal exposure to cocaine show exaggerated cerebrovascular responses to vasogenic stimuli and preserved cAMP-dependent cerebral vasoreactivity following asphyxia. PMID- 15337838 TI - Sudden onset of colitis after ablation of secretin-expressing lymphocytes in transgenic mice. AB - Though secretin mRNA was demonstrated in mouse lymphoid organs, its role in the immune system is unknown. Here, secretin gene-expressing cells were ablated by ganciclovir infusion in mice transgenic for the rat secretin promoter (Sec) directing the expression of herpesvirus thymidine kinase (Sec-HSVTK). Thymus, spleen, blood, and colon were investigated by histology. Lymphoid cells were extracted and quantified, and CD19+ B-cells and CD3+, CD103+, CD4+, and CD8+ T cells were analyzed by flow cytometry. Protein extracts from spleen and thymus were assayed for secretin by Western blotting, and isolated lymphocytes were investigated for HSVTK, secretin, and secretin receptor (Sec-R) mRNA by reverse transcription-polymerase chain reaction (RT-PCR). Ablation of secretin-expressing cells produced severe colitis with morphological features similar to those observed in graft-versus-host (GVH) disease. Profound lymphoid depletion was observed in spleen, thymus, and peripheral blood. The relative percentage of B- and T-cell subsets were unaffected. Analysis of colonic lymphocytes revealed a marked depletion of CD4+ T lymphocytes. Colitis and lymphoid depletion were not reversed by secretin cotreatment. Immunoblot analysis of protein extracts from spleen and thymus identified secretin-like immmunoreactant. RT-PCR of lymphocyte mRNA from spleen and thymus identified secretin and secretin receptor transcripts. We conclude that GVH-like colitis in ganciclovir-treated Sec-HSVTK mice arises from depletion of secretin gene-expressing lymphoid cells and not from the failure of secretin production. PMID- 15337839 TI - Dietary organic isothiocyanates are cytotoxic in human breast cancer MCF-7 and mammary epithelial MCF-12A cell lines. AB - Organic isothiocyanates (ITCs) are dietary components present in cruciferous vegetables. The purpose of this investigation was to examine the cytotoxicity of 1-naphthyl isothiocyanate (NITC), benzyl isothiocyanate (BITC), beta-phenethyl isothiocyanate (PEITC), and sulforaphane in human breast cancer MCF-7 and human mammary epithelium MCF-12A cell lines, as well as in a second human epithelial cell line, human kidney HK-2 cells. The cytotoxicity of NITC, BITC, PEITC, and sulforaphane, as well as the cytotoxicity of the chemotherapeutic agents daunomycin (DNM) and vinblastine (VBL), were examined in MCF-7/sensitive (wt), MCF-7/Adr (which overexpresses P-glycoprotein), MCF-12A, and HK-2 cells. Cell growth was determined by a sulforhodamine B assay. The IC50 values for DNM and VBL in MCF-7/Adr cells were 7.12 +/- 0.42 microM and 0.106 +/- 0.004 microM (mean +/- SE) following a 48-hr exposure; IC50 values for BITC, PEITC, NITC, and sulforaphane were 5.95 +/- 0.10, 7.32 +/- 0.25, 77.9 +/- 8.03, and 13.7 +/- 0.82 microM, respectively, with similar values obtained in MCF-7/wt cells. Corresponding values for BITC, PEITC, NITC, and sulforaphane in MCF-12A cells were 8.07 +/- 0.29, 7.71 +/- 0.07, 33.6 +/- 1.69, and 40.5 +/- 1.25 microM, respectively. BITC and PEITC can inhibit the growth of human breast cancer cells as well as human mammary epithelium cells at concentrations similar to those of the chemotherapeutic drug DNM. Sulforaphane and NITC exhibited higher IC50 values. The effect of these ITCs on cell growth may contribute to the cancer chemopreventive properties of ITCs by suppressing the growth of preclinical tumors, and may indicate a potential use of these compounds as chemotherapeutic agents in cancer treatment. PMID- 15337840 TI - Nutrition- and virus-induced stress represses the expression of manganese superoxide dismutase in vitro. AB - The relationship between oxidative stress and neuronal cell death has been suggested for many years. To understand the influence of oxidative stress on neuronal cell death, we investigated the influence of oxidative stress on DEV cells, a human glial cell line. Using enterovirus infection and/or malnutrition to induce oxidative stress, our results demonstrate that those stressors severely influence the antioxidant defense system in DEV cells. Although the expression of mitochondrial manganese superoxide dismutase (MnSOD) in DEV cells was significantly increased in acute infection with viral and nutritional stress, in persistent infection and nutritional stress, the expression of the MnSOD was drastically downregulated. We believe that this downregulation of MnSOD expression in the chronic stress model is due to repression of antioxidant defense. The downregulation of the MnSOD expression may lead to an increase of free-radical production and thus explain why the cells in the chronic stress model were more vulnerable to other oxidative stress influences. The vulnerability of DEV cells to additional stress factors resulted in progressive cell death, which may be analogous to the cell death in neurodegenerative diseases. PMID- 15337841 TI - Protein kinase B, P34cdc2 kinase, and p21 ras GTP-binding in kidneys of aging rats. AB - Renal nephropathy present in male Wistar rats more than 13 months of age was reported as an indication that the rats were in renal failure. In this study, the renal tissue damage at 14 months of age in male Munich Wistar rats was similar to that reported for Wistar rats, indicating that Munich Wistar rats could be another model for study of kidney function in the aging rat. The usual renal response to injury involves increased cell division and/or reparative processes that involve tyrosine kinase activity (TyrK) and/or guanosine triphosphate binding (G) protein signal trans-duction pathways. This study reveals the presence of renal tissue damage coinciding with significantly reduced activity of Ras, Akt, and p34cdc2 kinase, the signaling proteins that regulate cell division and/or growth, in renal cortical tissues of aging rats compared to young rats (P < 0.005, P < 0.005, and P< 0.001, respectively). These results suggest that proteins involved in signal transduction pathways associated with cell replication are downregulated in the aging kidney cortex at a time when renal cellular damage is also present. PMID- 15337842 TI - Tetrathiomolybdate therapy protects against concanavalin a and carbon tetrachloride hepatic damage in mice. AB - Tetrathiomolybdate, an anticopper drug, has been shown to protect mice against pulmonary fibrosis from bleomycin. Our hypothesis is that it does so by inhibiting fibrosis-inducing cytokines. Indeed, we have good evidence, not yet published, that tetrathiomolybdate inhibits pulmonary levels of transforming growth factor-beta and tumor necrosis factor-alpha expression in these bleomycin experiments. Herein, we evaluate tetrathiomolybdate's effectiveness in mitigating hepatitis and fibrosis in mice from the hepatotoxins, concanavalin A and carbon tetrachloride, and its inhibition of cytokines as a possible mechanism. In short term experiments, concanavalin A elevated serum amino leucine transferase levels several fold, and tetrathiomolybdate completely prevented this increase. In additional experiments, tetrathiomolybdate therapy reversed the elevated serum transaminase levels despite continued concanavalin A injections, with nearly significant serum interleukin-1beta inhibition. Concanavalin A given for 12 weeks produced mild fibrosis, whereas concomitant tetrathiomolybdate treatment resulted in normal histology. Carbon tetrachloride given for 12 weeks resulted in very high serum amino leucine transferase levels, high serum transforming growth factor-beta levels, cirrhosis as seen histologically, and increase in liver hydroxyproline, a measure of fibrosis. Concomitant tetrathiomolybdate partially and significantly protected against increases in amino leucine transferase and transforming growth factor-beta, fully protected against the increase in hydroxyproline, and resulted in normal histology. In conclusion, tetrathiomolybdate protects against the hepatitis and fibrosis produced by these hepatotoxins, probably by inhibiting the excessive increase in inflammatory and fibrotic cytokines. PMID- 15337843 TI - In search of RNase P RNA from microbial genomes. AB - A simple procedure has been developed to quickly retrieve and validate the DNA sequence encoding the RNA subunit of ribonuclease P (RNase P RNA) from microbial genomes. RNase P RNA sequences were identified from 94% of bacterial and archaeal complete genomes where previously no RNase P RNA was annotated. A sequence was found in camelpox virus, highly conserved in all orthopoxviruses (including smallpox virus), which could fold into a putative RNase P RNA in terms of conserved primary features and secondary structure. New structure features of RNase P RNA that enable one to distinguish bacteria from archaea and eukarya were found. This RNA is yet another RNA that can be a molecular criterion to divide the living world into three domains (bacteria, archaea, and eukarya). The catalytic center of this RNA, and its detection from some environmental whole genome shotgun sequences, is also discussed. PMID- 15337844 TI - Photolabile anticodon stem-loop analogs of tRNAPhe as probes of ribosomal structure and structural fluctuation at the decoding center. AB - With the recent availability of high-resolution structures of bacterial ribosomes, studies of ribosome-catalyzed protein biosynthesis are now focusing on the nature of conformational changes that occur as the ribosome exerts its complex catalytic function. Photocrosslinking can be relevant for this purpose by providing clues to ribosomal structural fluctuations and dynamics. Here we describe crosslinking experiments on 70S ribosomes using two photolabile anticodon stem-loop derivatives of Escherichia coli tRNAPhe carrying a 4 thiouridine in either position 33 or 37 and denoted Ph-ASLs. One or both of these Ph-ASLs bind to the tRNA A-, P-, and E-sites on the ribosome, with both binding to and photocrosslinking from the E-site showing strong dependence on the presence of a tRNA in the P-site. Both Ph-ASLs crosslink to the extreme 3'-end of 16S rRNA from both the P- and E-sites, providing direct confirmatory evidence in solution for the folding back of the 3'-end toward the decoding region. This suggests that the 3'-end of 16S rRNA may act as a switch in controlling mRNA access to the decoding center, a phenomenon of potential relevance for the translation of leaderless mRNA. E-site bound Ph-ASLs also form photocrosslinks to nucleotides 1395-1398, 1399-1400, and 1491-1494 at the top of helix 44 of 16S rRNA, indicating movement of the decoding center from a position between the A- and P-sites seen in the crystal structure to one neighboring the E-site. PMID- 15337845 TI - Intracellular folding of the Tetrahymena group I intron depends on exon sequence and promoter choice. AB - The Tetrahymena group I intron splices 20 to 50 times faster in Tetrahymena than in vitro, implying that the intron rapidly adopts its active conformation in the cell. The importance of cotranscriptional folding and the contribution of the rRNA exons to the stability of the active pre-RNA structure were investigated by comparing the activity of minimal pre-RNAs expressed in Escherichia coli. Pre RNAs containing exons derived from E. coli 23 S rRNA were three to four times more active than the wild-type Tetrahymena pre-RNA. E. coli transcripts of the chimeric E. coli pre-RNA were two to eight times more active than were T7 transcripts. However, the effect of cotranscriptional folding depends on exon sequences. Unexpectedly, the unspliced pre-RNA decays more slowly than predicted from the rate of splicing. This observation is best explained by partitioning of transcripts into active and inactive pools. We propose that the active pool splices within a few seconds, whereas the inactive pool is degraded without appreciable splicing. PMID- 15337846 TI - A cotranscriptional model for 3'-end processing of the Saccharomyces cerevisiae pre-ribosomal RNA precursor. AB - Cleavage of the Saccharomyces cerevisiae primary ribosomal RNA (rRNA) transcript in the 3' external transcribed spacer (ETS) by Rnt1p generates the 35S pre-rRNA, the earliest detectable species in the pre-rRNA processing pathway. In this study we show that Rnt1p is concentrated in a subnucleolar dot-shaped territory distinct from the nucleolar body. The 35S pre-rRNA is localized at the periphery of the Rnt1p dot, in a pattern that suggests a diffusion of the 35S pre-rRNA from the site of Rnt1p processing. When plasmid-borne versions of the rDNA are used to express rRNAs, the Rnt1p territory reorganizes around these plasmids, suggesting a close association between Rnt1p and the plasmid-borne rDNA units. Rnt1p was found associated with the endogenous rDNA by chromatin immunoprecipitation. Deletion of functionally important Rnt1p domains result in a loss of the dot shaped territory, showing that this subnucleolar territory corresponds to a functional site of processing. These results show that a large fraction of Rnt1p is localized at the site of transcription of the rDNA, suggesting that the cleavage of the primary pre-rRNA transcript to generate the 35S pre-rRNA is a cotranscriptional event. PMID- 15337847 TI - Ionic interactions between PRNA and P protein in Bacillus subtilis RNase P characterized using a magnetocapture-based assay. AB - Ribonuclease P (RNase P) is a ribonucleoprotein complex that catalyzes the cleavage of the 5' end of precursor tRNA. To characterize the interface between the Bacillus subtilis RNA (PRNA) and protein (P protein) components, the intraholoenzyme KD is determined as a function of ionic strength using a magnetocapture-based assay. Three distinct phases are evident. At low ionic strength, the affinity of PRNA for P protein is enhanced as the ionic strength increases mainly due to stabilization of the PRNA structure by cations. Lithium substitution in lieu of potassium enhances the affinity at low ionic strength, whereas the addition of ATP, known to stabilize the structure of P protein, does not affect the affinity. At high ionic strength, the observed affinity decreases as the ionic strength increases, consistent with disruption of ionic interactions. These data indicate that three to four ions are released on formation of holoenzyme, reflecting the number of ion pairs that occur between the P protein and PRNA. At moderate ionic strength, the two effects balance so that the apparent KD is not dependent on the ionic strength. The KD between the catalytic domain (C domain) and P protein has a similar triphasic dependence on ionic strength. Furthermore, the intraholoenzyme KD is identical to or tighter than that of full-length PRNA, demonstrating that the P protein binds solely to the C domain. Finally, pre-tRNAasp (but not tRNAasp) stabilizes the PRNA*P protein complex, as predicted by the direct interaction between the P protein and pre-tRNA leader. PMID- 15337848 TI - Recruitment of the Puf3 protein to its mRNA target for regulation of mRNA decay in yeast. AB - The Puf family of RNA-binding proteins regulates mRNA translation and decay via interactions with 3' untranslated regions (3' UTRs) of target mRNAs. In yeast, Puf3p binds the 3' UTR of COX17 mRNA and promotes rapid deadenylation and decay. We have investigated the sequences required for Puf3p recruitment to this 3' UTR and have identified two separate binding sites. These sites are specific for Puf3p, as they cannot bind another Puf protein, Puf5p. Both sites use a conserved UGUANAUA sequence, whereas one site contains additional sequences that enhance binding affinity. In vivo, presence of either site partially stimulates COX17 mRNA decay, but full decay regulation requires the presence of both sites. No other sequences outside the 3' UTR are required to mediate this decay regulation. The Puf repeat domain of Puf3p is sufficient not only for in vitro binding to the 3' UTR, but also in vivo stimulation of COX17 mRNA decay. These experiments indicate that the essential residues involved in mRNA decay regulation are wholly contained within this RNA-binding domain. PMID- 15337849 TI - Tethering of human Ago proteins to mRNA mimics the miRNA-mediated repression of protein synthesis. AB - MicroRNAs (miRNAs) are approximately 21-nt-long RNAs involved in regulating development, differentiation, and other processes in eukaryotes. In metazoa, nearly all miRNAs control gene expression by imperfectly base-pairing with the 3' untranslated region (3'-UTR) of target mRNAs and repressing protein synthesis by an unknown mechanism. It is also unknown whether miRNA-mRNA duplexes containing mismatches and bulges provide specific features that are recognized by factors mediating the repression. miRNAs form part of ribonucleoprotein complexes, miRNPs, that contain Argonaute (Ago) and other proteins. Here we demonstrate that effects of miRNAs on translation can be mimicked in human HeLa cells by the miRNA independent tethering of Ago proteins to the 3'-UTR of a reporter mRNA. Inhibition of protein synthesis occurred without a change in the reporter mRNA level and was dependent on the number, but not the position, of the hairpins tethering hAgo2 to the 3'-UTR. These findings indicate that a primary function of miRNAs is to guide their associated proteins to the mRNA. PMID- 15337851 TI - Apoptosis and apoptosis-related proteins in inflammatory bowel disease. PMID- 15337850 TI - Trans-splicing and polyadenylation of let-7 microRNA primary transcripts. AB - Members of the microRNA (miRNA) class of 22-nucleotide RNAs regulate the expression of target genes that contain sequences of antisense complementarity. Maturation of miRNAs involves cleavage of longer primary transcripts, but little is yet understood about how miRNA genes are transcribed and enter the processing pathway. We find that relatively long, polyadenylated transcripts encoded by the Caenorhabditis elegans let-7 gene undergo trans-splicing to the spliced leader 1 (SL1) RNA. Deletions, including removal of the trans-splice site, upstream of mature let-7 sequence result in stable accumulation of primary transcripts and compromised production of mature let-7 RNA in vivo. Our data show that multiple steps of let-7 miRNA biogenesis can be uncoupled, allowing for complex regulation in the production of a functional miRNA. Finally, the observation that let-7 primary transcripts undergo splicing highlights the importance of identifying the sequence of endogenous pri-miRNA substrates recognized by the cellular processing machinery. PMID- 15337852 TI - Functional characterization of a Drosophila mitochondrial uncoupling protein. AB - Sequence alignment of conserved signature motifs predicts the existence of the uncoupling protein 5 (UCP5)/brain mitochondrial carrier protein (BMCP1) homologue in Drosophila melanogaster. Here we demonstrate the functional characterization of the Drosophila melanogaster UCP5 protein (DmUCP5) in the heterologous yeast system, the first insect UCP reported to date. We show that physiological levels of DmUCP5 expression are responsible for an increase in state 4 respiration rates and a decrease in mitochondrial membrane potential. Furthermore, similar to UCP1, UCP2, and UCP3, the uncoupling activity of DmUCP5 is augmented by fatty acids and inhibited by the purine nucleotide GDP. Thus, DmUCP5 shares the mechanisms known to regulate the UCPs characterized to date. A lack of growth inhibition observed in DmUCP5 expressing yeast is consistent with the notion that physiological uncoupling has a minimal effect on cell growth. Finally, semiquantitative RT-PCR analysis shows a distinctive pattern of DmUCP5 expression predominantly localized in the adult head, similar to the expression pattern of its mammalian homologues. The conserved regulation of the expression of this gene from mammals to fruit flies suggests a role for UCP5 in the brain. PMID- 15337853 TI - Upregulation of the mitochondrial phosphate carrier during freezing in the wood frog Rana sylvatica: potential roles of transporters in freeze tolerance. AB - Screening of a cDNA library prepared from liver of the freeze-tolerant wood frog (Rana sylvatica) identified a freeze-responsive clone containing a 1370-nt sequence with an open reading frame of 360 amino acids. Sequence analysis revealed 84-86% identity with the mammalian inorganic phosphate carrier (PiC) that spans the inner mitochondrial membrane. Northern blot analysis showed that pic transcript levels increased over a time course of freezing, reaching 60-fold upregulation after 24-h frozen. Transcript levels were also assessed under freezing-related stresses with results showing a strong increase in pic transcript levels in response to dehydration (elevated 9.0-fold in 40% dehydrated frogs) but not under anoxia. Western blotting revealed elevated PiC protein over a time course of freeze-thaw whereas other mitochondrial carriers (dicarboxylate carrier, oxoglutarate transporter) of the same family were not affected by freezing. This modulation of PiC protein levels may play a role in mitochondrial ionic and/or osmotic balance during freeze-induced cell volume reduction. PMID- 15337854 TI - Fatty acids induce chloride permeation in rat liver mitochondria by activation of the inner membrane anion channel (IMAC). AB - The inner membrane of freshly isolated mammalian mitochondria is poorly permeable to Cl(-). Low, nonlytic concentrations (< or =30 microM) of long-chain fatty acids or their branched-chain derivatives increase permeation of Cl(-) as indicated from rapid large-scale swelling of mitochondria suspended in slightly alkaline KCl medium (supplemented with valinomycin). Myristic, palmitic, or 5 doxylstearic acid are powerful inducers of Cl(-) permeation, whereas lauric, phytanic, stearic, or 16-doxylstearic acid stimulate Cl(-) permeation in a lesser extent. Fatty acid-induced Cl(-) permeation across the inner membrane correlates well with the property of nonesterified fatty acids to release endogenous Mg(2+) from mitochondria. Myristic acid stimulates anion permeation in a selective manner, similar as was described for A23187, an activator of the inner membrane anion channel (IMAC). Myristic acid-induced Cl(-) permeation is blocked by low concentrations of tributyltin chloride (IC(50) approximately 1.5 nmol/mg protein). Moreover, myristic acid activates a transmembrane ion current in patch clamped mitoplasts (mitochondria with the outer membrane removed) exposed to alkaline KCl medium. This current is best ascribed to the opening of an ion channel with a single-channel conductance of 108 pS. We propose that long-chain fatty acids can activate IMAC by withdrawal of Mg(2+) from intrinsic binding sites. PMID- 15337855 TI - Ligand-dependent structural changes in the V(1) ATPase from Manduca sexta. AB - The response of V(1) ATPase of the tobacco hornworm Manduca sexta to Mg(2+) and nucleotide binding in the presence of the enhancer methanol has been studied by CuCl(2)-induced disulfide formation, fluorescence spectroscopy, and small-angle X ray scattering. When the V(1) complex was supplemented with CuCl(2) nucleotide dependence of A-B-E and A-B-E-D cross-linking products was observed in absence of nucleotides and presence of MgADP+Pi but not when MgAMP.PNP or MgADP were added. A zero-length cross-linking product of subunits D and E was formed, supporting their close proximity in the V(1) complex. The catalytic subunit A was reacted with N-4[4-[7-(dimethylamino)-4-methyl]coumarin-3-yl]maleimide (CM) and spectral shifts and changes in fluorescence intensity were detected upon addition of MgAMP.PNP, -ATP, -ADP+Pi, or -ADP. Differences in the fluorescence emission of these nucleotide-binding states were monitored using the intrinsic tryptophan fluorescence. The structural composition of the V(1) ATPase from M. sexta and conformational alterations in this enzyme due to Mg(2+) and nucleotide binding are discussed on the basis of these and previous observations. PMID- 15337856 TI - Overexpression of the inhibitor protein IF(1) in AS-30D hepatoma produces a higher association with mitochondrial F(1)F(0) ATP synthase compared to normal rat liver: functional and cross-linking studies. AB - According to functional studies, the higher IF(1) content reported in mitochondria of cancerous cells is supposed to induce a higher association with the F(1)F(0) complex than in normal cells and therefore a better inhibition of its ATPase activity. The first structural evidence supporting this prediction is here presented. Densitometric analyses of Western blotting experiments indicated a 2-fold increase in IF(1) content of AS-30D submitochondrial particles compared to normal rat liver controls. The ratio of IF(1)/F(1) alpha subunit increased similarly as judged by Westernblot analyses. This IF(1) overexpression correlated with a slower rate of IF(1) release (F(1)F(0)-ATPase activation) from the F(1)F(0) complex in AS-30D than in normal rat liver submitochondrial particles. The IF(1)-IF(1), gamma-IF(1), and alpha-IF(1) cross-linkages previously formed with dithiobis(succinimidylpropionate) in bovine F(1)F(0)I and IF(1) complexes were reproduced in the F(1)F(0)I-ATP synthase of hepatoma AS-30D cells. However, a much lower yield of IF(1) cross-linkages was found in normal rat liver particles which made them almost undetectable in SMP as well as in the immunoprecipitated F(1)F(0)I complex. Modeling in vivo IF(1) overexpression of cancerous cells by in vitro reconstitution of excess recombinant IF(1) with rat liver submitochondrial particles devoid of IF(1) reproduced the same IF(1) cross linkages observed in AS-30D particles. PMID- 15337857 TI - Functional approach to the catalytic site of the sarcoplasmic reticulum Ca(2+) ATPase: binding and hydrolysis of ATP in the absence of Ca(2+). AB - Isolated sarcoplasmic reticulum vesicles in the presence of Mg(2+) and absence of Ca(2+) retain significant ATP hydrolytic activity that can be attributed to the Ca(2+)-ATPase protein. At neutral pH and the presence of 5 mM Mg(2+), the dependence of the hydrolysis rate on a linear ATP concentration scale can be fitted by a single hyperbolic function. MgATP hydrolysis is inhibited by either free Mg(2+) or free ATP. The rate of ATP hydrolysis is not perturbed by vanadate, whereas the rate of p-nitrophenyl phosphate hydrolysis is not altered by a nonhydrolyzable ATP analog. ATP binding affinity at neutral pH and in a Ca(2+) free medium is increased by Mg(2+) but decreased by vanadate when Mg(2+) is present. It is suggested that MgATP hydrolysis in the absence of Ca(2+) requires some optimal adjustment of the enzyme cytoplasmic domains. The Ca(2+)-independent activity is operative at basal levels of cytoplasmic Ca(2+) or when the Ca(2+) binding transition is impeded. PMID- 15337858 TI - Adolescent outcome of ADHD: impact of childhood conduct and anxiety disorders. AB - OBJECTIVE: This study examines the impact of comorbidity of attention deficit/hyperactivity disorder (ADHD) with disruptive and anxiety disorders in childhood on clinical course and outcome. We consider the relative contribution of each comorbid symptom constellation, and also their interaction, to assess the following questions: (1) Does early comorbidity with conduct disorder (CD) and anxiety disorders define specific developmental trajectories?; (2) Is comorbid anxiety disorders in childhood continuous with anxiety disorders in adolescence?; (3) Does comorbid anxiety disorders mitigate the negative behavioral outcome of youth with ADHD?; and (4) Is there an interaction between comorbid CD and anxiety disorders, when they occur simultaneously, that predicts a different outcome than either comorbid condition alone? METHOD: Thirty-two 15- to 18-year-old adolescent males, diagnosed with ADHD between 7 and 11 years of age, were re-evaluated for assessment of adolescent outcome 4.3-9.2 years later. Hierarchical regression analyses were run with each of the eight Child Behavior Checklist and Youth Self Report problem scales, and the four anxiety symptom subscales of the Multidimensional Anxiety Scale for Children serving as outcome variables. RESULTS: Findings indicate that comorbid CD at baseline predicted parent reports of behavior problems in adolescence, while comorbid anxiety disorders in childhood predicted youth reports of anxiety and social problems. Anxiety disorders without CD did not predict poor behavioral outcome. Children with both comorbid CD and anxiety disorder had the highest levels of parent-rated symptoms on follow up. In particular, adolescent social problems were best predicted by the combination of comorbid CD and anxiety disorder in childhood. CONCLUSION: These data provide evidence that children with ADHD plus anxiety disorder do in fact have anxiety disorders, and that the combination of anxiety disorder and CD predicts a more rather than less severe course. PMID- 15337859 TI - Discerning disorders and diseases. PMID- 15337860 TI - ADHD: current questions and research. PMID- 15337861 TI - Self-report and objective measures of ADHD-related behaviors in parents of preschool children at risk for ADHD. AB - OBJECTIVE: Few studies have used a combination of objective and self-report measures to examine neuropsychological and behavioral functioning in parents of children with attention-deficit/hyperactivity disorder (ADHD). This study examined attention and inhibitory control in the parents of preschool children who were rated as "at risk" for developing ADHD as compared with parents of controls. METHODS: Preschool children (N=53) were divided into at risk for ADHD and control groups based on parent and teacher ratings of Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition ADHD symptoms. One parent of each child was administered an identical pairs Continuous Performance Test (CPT-IP), a Go/No-Go task, and the Brown Attention-Deficit Disorder Scale for Adults. RESULTS: Parents of preschoolers at risk for ADHD showed a pattern of responding on measures of vigilance characterized by slower reaction times and increased commission errors as compared with parents of controls. There were no significant group differences on self-report measures on the Brown Attention Deficit Disorder Scale for Adults. CONCLUSION: Parents of preschoolers at risk for ADHD appear to exhibit cognitive processing deficits that may not be evident using self-report measures. Further research is needed to more clearly identify the specific nature of these neuropsychological deficits and to determine whether they have a negative impact on their children. PMID- 15337862 TI - The utility of self-report measures and the continuous performance test in the diagnosis of ADHD in adults. AB - BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) occurs in as many as 4 percent of adults yet it is often not recognized in clinical settings because the presenting symptoms may resemble those seen in other disorders or because symptoms may be masked by commonly comorbid conditions such as anxiety and depression. OBJECTIVE: The purpose of this study was to examine the diagnostic utility of instruments commonly used in the assessment of adults presenting with symptoms of ADHD. METHODS: We reviewed several widely used self-report and laboratory measures and empirically examined the utility of the Brown Attention Deficit Disorder Scale for Adults (Brown ADD Scale) and the Conners Continuous Performance Test (CPT) in differentially identifying adults with ADHD and those with other Axis I disorders. RESULTS: Ninety-three adults who self-referred to the ADHD program for adults at a university medical center participated in the study. Of these, 44 had ADHD combined subtype (ADHD-CB), and 26 had ADHD, predominantly inattentive subtype (ADHD-IA). Thirty-three non-ADHD adults diagnosed with Axis I mood or anxiety disorders comprised an "Other Psychiatric" group. Rates of comorbid disorders, including substance abuse, in the ADHD groups were typical of those reported in the adult ADHD literature. Data on the Brown ADD Scale and on the CPT were available for subsets of 61 and 46 participants, respectively. Analyses showed that the ADHD-CB, ADHD-IA, and Other Psychiatric groups all received mean scores in the clinical range on the Brown ADD Scale, with a trend toward even higher elevations in the two ADHD groups. Among 12 CPT variables assessed for the three groups, the mean scores on only two variables for the ADHD-IA group were clinically elevated. Neither the Brown ADD Scale nor CPT scores evinced sufficient sensitivity and specificity to qualify them to assist in differential diagnosis of ADHD vis-a-vis other, predominantly internalizing, psychiatric disorders. CONCLUSION: The results indicate a need for closer examination of executive and adaptive functioning in adults with ADHD compared with those with internalizing disorders in order to identify features that could assist in differential diagnosis. PMID- 15337863 TI - The effect of childhood conduct disorder and cognitive functioning on adolescent substance use. AB - OBJECTIVE: To assess the impact of childhood conduct disorder (CD) and intelligence quotient (IQ) on later substance use in adolescence. METHODS: Neuropsychological and structured diagnostic evaluations were initially administered to 32 children with disruptive behavior disorder when they were 7-11 years of age. They were then re-evaluated on average 6.7 years later using an array of interviews and rating scales with a focus on substance use. RESULTS: Early CD and IQ scores together accounted for a significant proportion of the variance in later substance use (R2=.248). In addition, there was a significant CD and Verbal IQ interaction (R2=.164) such that high Verbal IQ was linked to increased substance use in adolescents who had childhood CD. CONCLUSION: These data indicate that the presence of conduct disorder may interact with high Verbal IQ during childhood in such a way as to predict later adolescent substance use in disruptive behavior disorder populations. PMID- 15337864 TI - Freeze, flight, fight, fright, faint: adaptationist perspectives on the acute stress response spectrum. AB - This article reviews the existing evolutionary perspectives on the acute stress response habitual faintness and blood-injection-injury type-specific phobia (BIITS phobia). In this article, an alternative evolutionary perspective, based on recent advances in evolutionary psychology, is proposed. Specifically, that fear-induced faintness (eg, fainting following the sight of a syringe, blood, or following a trivial skin injury) is a distinct Homo sapiens-specific extreme stress survival response to an inescapable threat. The article suggests that faintness evolved in response to middle paleolithic intra-group and inter-group violence (of con-specifics) rather than as a pan-mammalian defense response, as is presently assumed. Based on recent literature, freeze, flight, fight, fright, faint provides a more complete description of the human acute stress response sequence than current descriptions. Faintness, one of three primary physiological reactions involved in BIITS phobia, is extremely rare in other phobias. Since heritability estimates are higher for faintness than for fears or phobias, the author suggests that trait-faintness may be a useful complement to trait-anxiety as an endophenotype in research on the human fear circuitry. Some implications for the forthcoming Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition as well as for clinical, health services, and transcriptomic research are briefly discussed. PMID- 15337865 TI - First experience in human beings with a permanently implantable intrasac pressure transducer for monitoring endovascular repair of abdominal aortic aneurysms. AB - OBJECTIVES: Endovascular stent graft repair of abdominal aortic aneurysms (AAAs) prevents rupture by excluding the aneurysm sac from systemic arterial pressure. Current surveillance protocols after endovascular aneurysm repair (EVAR) follow secondary markers of sac pressurization, namely, endoleak and sac enlargement. We report the first clinical experience with the use of a permanently implantable, ultrasound-activated remote pressure transducer to measure intrasac pressure after EVAR. METHODS: Over 7 months, 14 patients underwent EVAR of an infrarenal abdominal aortic aneurysm with implantation of an ultrasound-activated remote pressure transducer fixed to the outside of the stent graft and exposed to the excluded aortic sac. Twelve patients received modular bifurcated stent grafts, and 2 patients received aortouniiliac devices. Intrasac pressures were measured directly with an intravascular catheter and by the remote sensor at stent-graft deployment. Follow-up sac pressures were measured with a remote sensor and correlated with systemic arterial pressure at every follow-up visit. Mean follow up was 2.6 +/-1.9 months. RESULTS: Excellent concordance was found between catheter-derived and transducer-derived intrasac pressssure intraoperatively. Pulsatile waveforms were seen in all functioning transducers at each evaluation interval. One implant ceased to function at 2 months of follow-up. In 1 patient a type I endoleak was diagnosed on 1-month computed tomography (CT) scans; 3 type II endoleaks were observed. Those patients with complete exclusion of the aneurysm on CT scans had a significant difference in systemic and sac systolic pressures initially (P <.001) and at 1 month (P <.001). Initial sac diastolic pressures were higher than systemic diastolic pressures (P <.001). The ratio of systemic to sac systolic pressure increased over time in those patients with complete aneurysm exclusion ( P <.001). Four of 6 patients with no endoleak and greater than 1-month follow-up had diminution of sac systolic pressure to 40 mm Hg or less by 3 months. CONCLUSION: This is the first report of a totally implantable chronic pressure transducer to monitor the results of EVAR in human beings. Aneurysm exclusion leads to gradual diminution of sac pressure over several months. Additional clinical follow-up will be necessary to determine whether aneurysm sac pressure monitoring can replace CT in the long-term surveillance of patients after EVAR. PMID- 15337866 TI - Overt ischemic colitis after endovascular repair of aortoiliac aneurysms. AB - OBJECTIVE: Controversy exists as to the cause of ischemic colitis complicating endovascular aneurysm repair. Occlusion of the hypogastric arteries (HAs) during endovascular repair of aortoiliac aneurysms (AIAs) results in a significant incidence of buttock claudication, and has been suggested as a causative factor in the development of postprocedural colonic ischemia, in addition to factors such as systemic hypotension, embolization of atheromatous debris, and interruption of inferior mesenteric artery inflow. To analyze the relationship between perioperative HA occlusion and postoperative ischemic colitis, we reviewed our experience over 2 years with Food and Drug Administration-approved endovascular graft devices for treatment of AIAs. METHODS: Elective repair of AIAs with bifurcated endovascular grafts was performed in 233 patients over a 2 year period. These included 184 AneuRx grafts, 17 Ancure grafts, and 32 Excluder grafts. During the experience, 44 patients (18.9%) underwent unilateral perioperative HA occlusion (28 right, 16 left) during the course of endovascular AIA repair, and 1 patient (0.4%) underwent bilateral HA occlusion. RESULTS: In 4 patients (1.7%) signs and symptoms of ischemic colitis developed 2.0 +/- 1.4 days postoperatively. In all patients the diagnosis was confirmed at sigmoidoscopy, and initial treatment included bowel rest, hydration, and intravenous antibiotic agents. Three patients with bilateral patent HAs required colonic resection 14.7 +/- 9.7 days after the initial diagnosis, and 2 of these 3 patients died in the postoperative period. Pathologic findings confirmed the presence of atheroemboli in the colonic vasculature in all 3 patients who underwent colonic resection. The fourth patient had undergone multiple manipulations of the left HA in an unsuccessful attempt to preserve patency of this vessel during AIA repair. This patient recovered completely with nonoperative management. Perioperative unilateral HA occlusion was not associated with a significantly higher incidence of postoperative ischemic colitis. CONCLUSION: Perioperative HA occlusion during aortoiliac open or endovascular surgery may contribute to development of the rare but potentially lethal complication of ischemic colitis. However, our extensive experience suggests that embolization of atheromatous debris to the HA tissue beds during endovascular manipulations, rather than proximal HA occlusion, is the primary cause of clinically significant ischemic colitis after endovascular aneurysm repair. PMID- 15337867 TI - Initial management and outcome of aortic endograft limb occlusion. AB - OBJECTIVE: The purpose of this study was to determine the differences in outcome related to initial management of aortic endograft limb occlusion (ELO). METHODS: During a 7-year period, 823 endovascular aneurysm repairs (EVARs) resulted in 25 ELOs in 22 patients. The initial management and outcome of these ELOs were reviewed. Median follow-up after ELO was 24.2 +/- 16.8 months. RESULTS: Initial EVARs included both unsupported unibody (n = 5) and supported modular (n = 17) devices. ELO was significantly more common in the unsupported unibody graft design (P <.024) and with extension of the graft limb to the external iliac artery (P <.001). ELO was managed with an endovascular approach (EVA), including some combination of mechanical thrombectomy (n = 8), angioplasty with or without stenting (n = 8), and thrombolysis (n = 2) in 12 patients and bypass procedures (femoral-femoral bypass, n = 11; axillofemoral bypass, n = 1; and aortofemoral bypass, n = 1) in 13. At 12-month follow-up, freedom from secondary procedures with EVA was 80.2 +/- 17.7% versus 53.2 +/-17.1% with extra-anatomic bypass (EB) (P = NS). Secondary patency was 100% with EVA and 80.6 +/- 14.4% with EB (P = NS). Of the 12 EVAs, there was 1 (8.3%) perioperative mortality with EVA and none with EB. EB failure was directly attributed to donor limb occlusion in 4 of 6 EVAs (67%), and when this occurred it resulted in bilateral lower extremity ischemia. Amputation was required in 2 of 12 (16.7%) EBs versus none of the 12 EVAs (P = NS). EVA never resulted in graft dislodgement or endoleak but did identify an underlying treatable cause in 8 of 12 (67%). CONCLUSION: Both EVA and EB are acceptable management strategies for ELO. The potential risk of graft dislodgement was not observed with an EVA. If EB is employed, assessment of the donor limb and treatment of any underlying lesions is advisable in an attempt to minimize future donor limb occlusion. PMID- 15337868 TI - The role of aortic stent grafting in the treatment of atheromatous embolization syndrome: results after a mean of 15 months follow-up. AB - BACKGROUND: Endovascular stent-graft (ESG) repair of abdominal aortic aneurysm (AAA) has emerged as an alternative to open surgery. The role of ESG in patients with challenging medical and anatomic problems remains an area of general debate. This study reviews an experience with stent grafts to treat patients with AAA and atheromatous embolization syndrome (AES) presenting with chronic distal embolization (CDE). METHODS: Over a 5-year period 660 patients with AAA were treated with aortic stent grafts. Patients with AAA and ischemic ulcerations or toe gangrene due to CDE despite palpable foot pulses were investigated for successful aneurysm exclusion, resolution of ischemic symptoms, complications and survival. Follow-up averaged 15.3 +/- 14.9 months (range, 1 to 60 months). RESULTS: Nineteen patients had AAA and manifestations of CDE. The population (16 males/3 females) had a mean age of 79 +/- 7 years and mean aneurysm diameter of 5.5 cm. Renal insufficiency was present in 5/19 (26 %). Ischemia presented as ischemic ulcers (16/19 [84.2%]) or toe gangrene (3/19 [15.8%]). Stent grafts included 6 aortouniiliac and 13 bifurcated devices. Exclusion was achieved in all but 2 patients who had type II lumbar endoleaks. At 30-day postoperative follow up, mortality was 0 % and resolution of CDE/ischemia was noted in 2 of 19 (10.5%) patients. Eight of 9 patients with follow-up of 1 year had complete resolution of their ischemic symptoms, with no recurrent manifestations of AES. Complications included progression of renal insufficiency over an 18-month period in 1 patient and an unstable expanding pararenal aortic neck in 1 patient. Foot ischemia persisted at 1 year in a patient with severe coexisting thoracic aortic disease despite successful AAA exclusion. Six (31.6%) patients died during a mean follow up of 15.3 months from causes unrelated to their AAA. CONCLUSION: On the basis of this experience, stent-graft repair of AAA and CDE may be an effective strategy to prevent future embolization. Recognition of coexisting thoracic aortic disease is essential. ESG does not address the extremely high morbidity and mortality from cardiovascular causes in this population. PMID- 15337869 TI - Effectiveness of coiling in the treatment of endoleaks after endovascular repair. AB - BACKGROUND: Persistent endoleaks are a common problem following endovascular repair of abdominal aortic aneurysms, and the best method of treatment has been an issue of debate. Some experimental evidence has suggested that coiling may not be an effective method because it allows transmission of pressure across the coils with continued expansion of the sac. We reviewed our experience with endoleak coiling to assess the degree of clinical success of this treatment. METHODS: A retrospective review of patients with type I or type II endoleaks treated solely by coiling over a 7-year period (1997-2003) was performed. All endoleaks had been observed for at least 6 months prior to intervention to detect spontaneous resolution. All coils were delivered by selective catheterization of the endoleaks. For Type II endoleaks, the branches were all coiled at their junction with the sac when feasible and the endoleak cavity was packed. Clinical success was defined as cessation of endoleak on follow-up computed tomography as well as no further aneurysmal growth (> or =5mm minor axis). RESULTS: Twenty eight patients had their endoleaks treated only with coils. There were 22 Ancure, 2 Excluder, 2 AneuRx, and 2 Lifepath endografts in this patient cohort. Procedural morbidity was 0%. Mean follow-up after coiling for all patients was 18 months (range, 1-60 months) while mean follow-up for patients with a type I endoleak was 24 months. Clinical success was achieved in 15 of 19 (79%) patients with type II endoleaks and 8 of 9 (89%) patients with type I. Three patients, all with type I endoleak, required more than 1 episode of coiling, while 2 others, both with type II lumbar endoleaks, required repeat angiography due to inability to access the leak during the first attempt. There were 2 proximal and 6 distal type I endoleaks (2 aortic, 6 iliac) successfully treated while the type II successes included 8 inferior mesenteric artery and 7 sole lumbar endoleaks. Five patients continued to show evidence of endoleak over time: 2 endoleaks were associated with aneurysm growth leading to conversion in 1 patient, 2 patients with type II endoleaks are stable, and the sole type I endoleak with continued perigraft flow has shown significant shrinkage of the sac and continues to be observed 18 months later. No ruptures were noted during follow-up. CONCLUSION: Coiling as the sole method of endoleak management may be a suitable treatment option in selected patients. Clinical success can be expected in over 80% of patients with type II and select type I endoleaks, with minimal morbidity. PMID- 15337870 TI - Midterm outcome of endovascular abdominal aortic aneurysm repair in octogenarians: a single institution's experience. AB - OBJECTIVES: We analyzed midterm durability of endovascular abdominal aortic aneurysm repair (EVAR) in octogenarians compared with younger patients. METHODS: Data for 182 consecutive patients who underwent elective EVAR between 1999 and 2003 were retrospectively reviewed. Forty-nine patients (27%) were 80 years or older (study group [SG]; mean age, 84 years; range, 80-89 years), and 133 patients (73%) were younger (control group [CG]; mean age, 72 years; range, 53-79 years). Chi2 analysis, Fisher exact test, Student t test, and Mann-Whitney U test were used as appropriate to test for intergroup differences. Kaplan-Meier curves, log-rank tests, and multivariate Cox models were used for time-to-event analysis, with P < or =.05 considered significant. RESULTS: Mean follow-up was 16 months (range, 1-43 months). Body weight was higher (P <.001), and elevated plasma lipid levels (59% vs 43%; P =.042) and use of nicotine (47% vs 29%; P =.015) more frequent in the octogenarians. Baseline aneurysm size, procedure-related data, and hospital stay were comparable between groups. Aneurysm-related mortality was 0% in the study group and 0.7% in the control group (P =.740). Systemic complications occurred in 22% (SG) versus 11% (CG) (P =.035), owing to a rise in serum creatinine concentration greater than 30% of baseline in 14% in the octogenarian group (vs 5% in the CG; P =.048). Groin lymphoceles developed in 12% (SG) versus 2% (CG; P =.013). Technical success was 96% (SG) versus 98% (CG; P =.408), and clinical success was 86% versus 90% (P =.269). No aneurysm rupture occurred during follow-up, and aneurysm-related adverse events were comparable between groups. The estimated risk for any type of endoleak (2.2; 95% confidence interval [CI], 1.1-4.2; P =.023) or type II endoleak (2.1; 95% CI, 1.0-4.3; P = 0.51) was higher in the study group versus the control group; however, this did not affect secondary procedure rates (SG 16% vs CG 12%; estimated risk, SG vs CG,: 1.5; 95% CI, 0.6-3.6; P = 0.420) or aneurysm remodeling (97.2% combined aneurysm sac stabilization or decrease in both groups; P =.592). Aneurysm enlargement occurred in 2.8% (SG 1 vs CG 4; P =.592). CONCLUSION: Elective EVAR in octogenarians appears safe and effective over midterm follow-up, with a temporary decrease in renal function (14%) and postoperative lymphoceles (12%) being the most common postoperative adverse events. Advanced chronologic age is not associated with diminished procedural outcome, clinical success, or postoperative survival, compared with younger age. Because of low perioperative mortality and high procedural success, EVAR may be the preferred approach to abdominal aortic aneurysm treatment in selected elderly patients. PMID- 15337871 TI - Is three-dimensional computed tomography reconstruction justified before endovascular aortic aneurysm repair? AB - OBJECTIVES: The endovascular management of abdominal aortic aneurysm (AAA) relies on accurate preoperative imaging for proper patient selection and operative planning. Three-dimensional (3-D) computed tomography (CT) with reformatted images perpendicular to blood flow has gained popularity as a method of AAA assessment and image-based planning before endovascular aneurysm repair (EVAR). The current study was undertaken to determine the interobserver agreement of AAA measurements obtained with axial CT and reformatted 3-D CT and to compare the consistency of the 2 methods in selecting patients for EVAR. METHODS: Eight observers assessed the axial CT and reformatted 3-D CT scans for 5 patients with AAAs to determine whether the patients were candidates for EVAR. 3-D CT with multiplanar reformatted images was performed by Medical Media Systems (MMS). Each observer measured the length and diameter of the proximal neck, maximal AAA, aortic bifurcation, common iliac diameter, and aortic angulation. The proximal neck and common iliac arteries were also assessed for thrombus, calcification, and tortuosity. Agreement of the measurements on axial CT scans was compared with those on MMS CT scans by calculating the kappa statistic. Complete agreement was defined as kappa = 1.0. The limits of agreement between observers were also calculated. RESULTS: The cumulative interobserver agreement of MMS CT scans (kappa =.81) was greater than for axial CT scans (kappa =.59). The kappa value for each of the diameter measurements was greater with the MMS CT scans. In 79% of cases the observers' measurements were less than 2 mm from the mean with MMS CT, compared with 59% for axial CT. The kappa value for deciding whether a patient was an endograft candidate on the basis of aortic neck was greater with the MMS CT (0.92 vs 0.63). The limits of agreement between observers were also better with the MMS CT. CONCLUSIONS: The interobserver agreement in planning EVAR is significantly better with MMS CT compared with traditional axial CT. The routine use of MMS CT appears justified before EVAR to improve the accuracy and consistency of patient selection. PMID- 15337872 TI - Total laparoscopic infrarenal aortic aneurysm repair: preliminary results. AB - OBJECTIVES: We describe our initial experience of total laparoscopic abdominal aortic aneurysm (AAA) repair. MATERIAL AND METHODS: Between February 2002 and September 2003, we performed 30 total laparoscopic AAA repairs in 27 men and 3 women. Median age was 71.5 years (range, 46-85 years). Median aneurysm size was 51.5 mm (range, 30-79 mm). American Society of Anesthesiologists class of patients was II, III and IV in 10, 19, and 1 cases, respectively. We performed total laparoscopic endoaneurysmorrhaphy and aneurysm exclusion in 27 and 3 patients, respectively. We used the laparoscopic transperitoneal left retrocolic approach in 27 patients. We operated on 2 patients via a tranperitoneal left retrorenal approach and 1 patient via a retroperitoneoscopic approach. RESULTS: We implanted tube grafts and bifurcated grafts in 11 and 19 patients, respectively. Two minilaparotomies were performed. In 1 case, exposure via a retroperitoneal approach was difficult and, in another case, distal aorta was extremely calcified. Median operative time was 290 minutes (range, 160-420 minutes). Median aortic clamping time was 78 minutes (range, 35-230 minutes). Median blood loss was 1680 cc (range, 300-6900 cc). In our early experience, 2 patients died of myocardial infarction. Ten major nonlethal postoperative complications were observed in 8 patients: 4 transcient renal insufficiencies, 2 cases of lung atelectasis, 1 bowel obstruction, 1 spleen rupture, 1 external iliac artery dissection, and 1 iliac hematoma. Others patients had an excellent recovery with rapid return to general diet and ambulation. Median hospital stay was 9 days (range, 8-37 days). With a median follow-up of 12 months (range, 0.5 20 months), patients had a complete recovery and all grafts were patent. CONCLUSION: These preliminary results show that total laparoscopic AAA repair is feasible and worthwhile for patients once the learning curve is overcome. However, prior training and experience in laparoscopic aortic surgery are needed to perform total laparoscopic AAA repair. Despite these encouraging results, a greater experience and further evaluation are required to ensure the real benefit of this technique compared with open AAA repair. PMID- 15337873 TI - Minimally invasive retroperitoneal approach for the treatment of infrarenal aortic disease. AB - PURPOSE: In order to decrease complications and improve postoperative recovery, we have developed a minimally invasive retroperitoneal approach (MIRPA) for the treatment of infrarenal aortic disease. This study was carried out to define the limitations and applicability of this technique in the treatment of aortoiliac occlusive disease (AIOD) and abdominal aortic aneurysms (AAAs). METHODS: From November 2000 to February 2004, 150 patients with AAA (n = 130) or AIOD (n = 20) were prospectively included in the study. The procedure consisted in a standard aneurysmorrhaphy or bypass procedure performed through a video assisted left minilombotomy.The main outcomes measured were mortality, complications, operative time, aortic cross-clamp time, time to solid diet, and length of intensive care unit and hospital stay. RESULTS: Operative mortality was 0.7 %. Nonfatal postoperative complications occurred in 12 patients (8%). Conversion to a standard procedure was necessary in 3 patients. Mean operative time was 207 +/- 57 minutes (AAA) and 224 +/- 55 minutes (AIOD). Mean aortic cross-clamp time was 76 +/- 26 minutes (AAA) and 48 +/- 21 minutes (AIOD). Median resumption of regular diet was 2 days. Median length of stay in the intensive care unit was 1 day and in the hospital 8 days. CONCLUSION: Our results suggest that MIRPA is a safe and effective minimally invasive procedure in the treatment of infrarenal aortic disease. PMID- 15337874 TI - Anti-platelet effect of aspirin is substantially reduced after administration of heparin during carotid endarterectomy. AB - OBJECTIVES: Aspirin therapy is usually continued throughout the perioperative period to reduce the risk for thromboembolic stroke and myocardial infarction after carotid endarterectomy (CEA). Aspirin irreversibly binds cyclooxygenase-1, thereby reducing platelet aggregation for the lifetime of each platelet. However, recent research from this unit has shown that aggregation in response to arachidonic acid increases significantly, but transiently, during CEA, which suggests that the anti-platelet effect of aspirin is temporarily reversed. The purpose of the current study was to determine when this phenomenon occurs and to identify the possible mechanisms involved. METHODS: Platelet aggregation was measured in platelet-rich plasma from 41 patients undergoing CEA who were stabilized with 150 mg of aspirin daily. Blood was taken at 8 time points: before anesthesia, after anesthesia, before heparinization, 3 minutes after heparinization, 3 minutes after shunt insertion, 10 minutes after flow restoration, 4 hours postoperatively, and 24 hours postoperatively. Platelet aggregation was also measured at similar times in a group of 18 patients undergoing peripheral angioplasty without general anesthesia. RESULTS: All patient platelets were effectively inhibited by aspirin at the start of the operation. There was a significant intraoperative increase in platelet response to arachidonic acid in both groups of patients, which occurred within 3 minutes of administration of unfractionated heparin. In the CEA group this resulted in a greater than 10-fold increase in mean aggregation, to 5 mmol/L of arachidonic acid (5 mmol/L), rising from 3.9% +/- 2.2% preoperatively to 45.1% +/- 29.3% after administration of heparin ( P <.0001). This increased aggregation persisted into the early postoperative period, but by 24 hours post operation aggregation had returned to near preoperative values. Aggregation in response to other platelet agonists (adenosine diphosphate, thrombin receptor agonist peptide) showed only a small increase at the same time, which could be accounted for by a parallel increase in the level of spontaneous aggregation. CONCLUSION: Administration of heparin significantly increases platelet aggregation in response to arachidonic acid, despite adequate inhibition by aspirin administered preoperatively. This apparent reversal in anti-platelet activity persisted into the immediate early postoperative period, and could explain why a small proportion of patients are at increased risk for acute cardiovascular events after major vascular surgery, despite aspirin therapy. PMID- 15337875 TI - Serum values of metalloproteinase-2 and metalloproteinase-9 as related to unstable plaque and inflammatory cells in patients with greater than 70% carotid artery stenosis. AB - OBJECTIVE: Unstable carotid plaques, characterized by increased levels of macrophages and T lymphocytes, have high emboligenic potential and carry a risk for producing cerebral ischemic events. It has been suggested that plaque instability may be mediated by the family of metalloproteinases (MMPs). The purpose of this study was to analyze the relationship between concentrations of MMP-2 and MMP-9 and unstable carotid plaques, presence of macrophages and T lymphocytes in the plaques, and neurologic symptoms, to establish additional risk markers in patients with greater than 70% carotid artery stenosis. This was a cross-sectional study carried out in a referral center and institutional practice in hospitalized patients. METHODS: The study included 40 patients with carotid artery stenosis treated with carotid endarterectomy. Of these patients, 67.5% had experienced a previous neurologic event and 32.5% exhibited no symptoms. MMP-2 and MMP-9 levels were determined with enzyme-linked immunosorbent assay 48 hours before surgery. Histopathologic analysis (stable or unstable) and immunohistochemistry (macrophage count, T lymphocytes, activated T lymphocytes) were carried out on the plaques. RESULTS: Mean MMP-2 and MMP-9 serum concentrations in the population studied were 1138.27 +/- 326.08 ng/mL and 1026.10 +/- 412.90 ng/mL, respectively. MMP-2 levels were significantly higher in patients with symptoms compared with patients without symptoms (1247.30 +/- 276.80 ng/mL vs 911.80 +/- 311.84 ng/mL; P =.001). MMP-9 was also significantly higher in the symptomatic group (1026.10 +/- 412.90 ng/mL vs 377.84 +/- 164.08 ng/mL; P =.001) and in patients with unstable plaques compared with those with stable plaques (1006.98 +/- 447.09 ng/mL vs 496.16 +/- 292.78 ng/mL; P =.001). In addition, we found a strong association between elevated MMP-9 concentration and presence of macrophages in plaque (Spearman rho, 0.45; P =.004). At logistic regression analysis, variables that best predicted the presence of unstable plaque were a previous neurologic event and MMP-9 level greater than 607 ng/mL (sensitivity, 96%; specificity, 92%; negative predictive value, 94.7%; positive predictive value, 93%). CONCLUSION: Elevated MMP-9 concentration is associated with carotid plaque instability and the presence of macrophages, factors that indicate increased risk for a neurological event. Determination of this gelatinase may enable identification of high-risk subgroups of patients with carotid artery stenosis. PMID- 15337876 TI - Transcervical carotid stenting with internal carotid artery flow reversal: feasibility and preliminary results. AB - OBJECTIVE: Transfemoral carotid artery stenting (CAS), with or without distal protection, is associated with risk for cerebral and peripheral embolism and access site complications. To establish cerebral protection before crossing the carotid lesion and to avert transfemoral access complications, the present study was undertaken to evaluate a transcervical approach for CAS with carotid flow reversal for cerebral protection. METHODS: Fifty patients underwent CAS through a transcervical approach. All patients with symptoms had greater than 60% internal carotid artery (ICA) stenosis, and all patients without symptoms had greater than 80% ICA stenosis. Twenty-one patients (42%) had symptomatic disease or ipsilateral stroke, and 8 patients (16%) had contralateral stroke. Four patients (8%) had recurrent stenosis, 7 patients (14%) had contralateral ICA occlusion, and 1 patient (2%) had undergone previous neck radiation. Twenty-seven procedures (54%) were performed with local anesthesia, and 23 (46%) with general anesthesia. Using a cervical cutdown, flow was reversed in the ICA by occluding the common carotid artery and establishing a carotid-jugular vein fistula. Pre-dilation was selective, and 8-mm to 10-mm self-expanding stents were deployed and post-dilated with 5-mm to 6-mm balloons in all cases. RESULTS: The procedure was technically successful in all patients, without significant residual stenoses. No strokes or deaths occurred. There was 1 wound complication (2%). All patients were discharged within 2 days of surgery. Mean flow reversal time was 21.4 minutes (range, 9-50 minutes). Carotid flow reversal was not tolerated in 2 patients (4%). Early in the experience, carotid flow reversal was not possible in 1 patient, and there were 1 major and 3 minor common carotid artery dissections, which resolved after stent placement. One intraoperative transient ischemic attack (2%) occurred in 1 patient in whom carotid flow was not reversed, and 1 patient with a contralateral ICA occlusion had a contralateral transient ischemic attack. At 1 to 12 months of follow-up, all patients remained asymptomatic, and all but 1 stent remained patent. CONCLUSION: Transcervical CAS with carotid flow reversal is feasible and safe. It can be done with the patient under local anesthesia, averts the complications of the transfemoral approach, and eliminates the increased complexity and cost of cerebral protection devices. Transcervical CAS is feasible when the transfemoral route is impossible or contraindicated, and may be the procedure of choice in a subset of patients in whom carotid stenting is indicated. PMID- 15337877 TI - Long-term results of covered stent repair of internal carotid artery dissections. AB - BACKGROUND: Traumatic and spontaneous dissections of internal carotid arteries (ICA) are rare conditions. So far, these pathologies are primarily treated conservatively, surgical revascularization being an option only after recurrent thromboembolic neurologic episodes or continuous aneurysm growth. Successful endovascular treatment strategies with covered stents have been reported in patients with ICA dissections. However, no long-term results are published so far. Herein, we report our experience of a combined conventional and endovascular repair of ICA dissections under reversed flow and their respective long-term results. METHODS: In a prospective evaluation of clinical and morphologic outcome of 6 patients with carotid artery dissections, 2 patients were treated for continuous aneurysm growth and 4 patients for high-grade ICA stenoses with recurrent thromboembolic episodes during a 6-month follow-up period. A 6-mm polytetrafluoroethylene Hemobahn endoprosthesis was inserted under reversed flow of the internal carotid artery. RESULTS: No perioperative strokes were observed; one TIA occurred, lasting less than 3 hours; no peripheral cranial nerve injuries or deaths were observed. No occlusions, hemodynamically significant stenosis, or recurrent neurologic symptoms were seen during follow-up, which ranged from 6 to 54 months (mean, 38.3 months). CONCLUSION: Open endovascular repair of the ICA of symptomatic patients with dissections with a 6-mm covered endoprosthesis is a safe alternative to conventional surgery, with excellent long-term patency. PMID- 15337878 TI - Impaired muscle oxygen use at onset of exercise in peripheral arterial disease. AB - OBJECTIVES: In patients with peripheral arterial disease (PAD), abnormal muscle metabolism and impaired oxygen delivery distal to the arterial occlusions may contribute to the exercise limitation observed in this population. Muscle tissue hemoglobin saturation (StO2), measured with near-infrared spectroscopy, reflects the relative contributions of oxygen delivery and oxygen use. Thus differences in the kinetics of StO2 in response to exercise may yield important insight into the potential mechanisms associated with the PAD exercise impairment. The purposes of this study were to characterize the muscle oxygenation responses in patients with PAD and in healthy control subjects at the onset of exercise, and to compare the kinetics of StO2 desaturation. We hypothesized that at the onset of exercise the kinetics of StO2 desaturation would be slowed in PAD compared with control responses. MATERIAL AND METHODS: Six patients with PAD and 6 healthy control subjects from a university center were examined in a prospective cross-sectional analysis that evaluated the desaturation kinetics of StO2 at the onset of walking exercise. On separate visits subjects performed graded treadmill exercise and 3 constant work rate treadmill tests equivalent to approximately 60% (low), approximately 80% (medium), and 100% (peak) of their peak exercise work rate. Gastrocnemious muscle StO2 response profiles (InSpectra tissue spectrometer) were measured at rest and across the rest to exercise transition. Muscle StO2 responses were characterized by an exponential mathematical model. The end point value was taken as the time constant of StO2 desaturation after onset of exercise (ie, equivalent to time to reach approximately 63% of StO2 decrease). RESULTS: The patients with PAD and the control subjects were of similar age and activity level. The qualitative patterns of StO2 responses at onset of exercise were also similar between patients and control subjects at all work rates. However, the kinetic time constants of StO2 desaturation were prolonged in patients with PAD versus control subjects (averaged time constant across all work rates, 21.9 +/- 9.4 seconds vs 4.9 +/- 2.2 seconds; P <.01). CONCLUSIONS: The slowed muscle StO2 kinetics in PAD are consistent with an impairment in muscle oxygen use at the onset of walking exercise. Impaired muscle metabolism may contribute to the altered physiologic responses to exercise and to exercise impairment in patients with PAD. PMID- 15337879 TI - Integrating surgery and radiology in one suite: a multicenter study. AB - PURPOSE: The study was performed to evaluate the performance of digital fixed mounted angiographic C-arm systems in the operating room as used by surgeons, cardiologists, and interventional radiologists. METHODS: An observational study in the operating room was performed, along with a structured questionnaire and semi-structured interviews. Twenty interventions were observed at 5 sites. Workflow was analyzed. RESULTS: Integration of high-end angiographic imaging equipment in the operating room enables image-guided surgery with high-quality images, on-table quality assessment of surgical procedures, and "one-stop shopping" procedures. Integrated suites were run by surgery as well as radiology departments, and are used for a variety of procedures, including vascular, cardiothoracic, open surgical, percutaneous, and combined procedures. Operation of the angiographic system and its user interface design were not considered ideal for operating room use. Limited patient accessibility was observed, sometimes leading to uncomfortable positions for the operating physicians. Certain procedures, such as tibial artery surgery, were difficult to perform, owing to lack of accessories. Patient transfer was considered inadequate. Cleaning of the system was rated as poor. Operating room use puts an even higher demand on reliability of the system. CONCLUSION: Integration of digital angiographic systems into operating rooms has produced opportunities for new treatments and offers a superior solution for interdisciplinary work among surgeons, cardiologists, and radiologists. However, the context of use differs radically from that in the traditional radiologic examination room; the environment, users, and procedures are all different. Integration of imaging methods into the operating room can be more successful if special operating room conditions are taken into account by medical systems manufacturers. PMID- 15337880 TI - Deep venous thrombosis after radiofrequency ablation of greater saphenous vein: a word of caution. AB - PURPOSE: Radiofrequency ablation (RFA) of the greater saphenous vein (GSV; "closure") is a relatively new option for treatment of venous reflux. However, our initial enthusiasm for this minimally invasive technique has been tempered by our preliminary experience with its potentially lethal complication, deep venous thrombosis (DVT). METHODS: Seventy-three lower extremities were treated in 66 patients with GSV reflux, between April 2003 and February 2004. There were 48 (73%) female patients and 18 (27%) male patients, with ages ranging from 26 to 88 years (mean, 62 +/- 14 years). RFA was combined with stab avulsion of varicosities in 55 (75%) patients and subfascial ligation of perforator veins in 6 (8%) patients. An ATL HDI 5000 scanner with linear 7-4 MHz probe and the SonoCT feature was used for GSV mapping and procedure guidance in all procedures. GSV diameter determined the size of the RFA catheter used. Veins less than 8 mm in diameter were treated with a 6F catheter (n = 54); an 8F catheter was used for veins greater than 8 mm in diameter (n = 19). The GSV was cannulated at the knee level. The tip of the catheter was positioned within 1 cm of the origin of the inferior epigastric vein (first GSV tributary). All procedures were carried out according to manufacturer guidelines. RESULTS: All patients underwent venous duplex ultrasound scanning 2 to 30 days (mean, 10 +/- 6 days) after the procedure. The duplex scans documented occlusion of the GSV in 70 limbs (96%). In addition, DVT was found in 12 limbs (16%). Eleven patients (92%) had an extension of the occlusive clot filling the treated proximal GSV segment, with a floating tail beyond the patent inferior epigastric vein into the common femoral vein. Another patient developed acute occlusive clots in the calf muscle (gastrocnemius) veins. Eight patients were readmitted and received anticoagulation therapy. Four patients were treated with enoxaparin on an ambulatory basis. None of these patients had pulmonary embolism. Initially 3 patients with floating common femoral vein clots underwent inferior vena cava filter placement. Of the 19 limbs treated with the 8F RFA catheter, GSV clot extension developed in 5 (26%), compared with 7 of 54 (13%) limbs treated with the 6F RFA catheter (P =.3). No difference was found between the occurrence of DVT in patients who underwent the combined procedure (RFA and varicose vein excision) compared with patients who underwent GSV RFA alone (P =.7). No statistically significant differences were found in age or gender of patients with or without postoperative DVT (P = NS). CONCLUSION: Patients who underwent combined GSV RFA and varicose vein excision did not demonstrate a higher occurrence of postoperative DVT compared with patients who underwent RFA alone. Early postoperative duplex scans are essential, and should be mandatory in all patients undergoing RFA of the GSV. PMID- 15337881 TI - Side-to-side sutureless vascular anastomosis with magnets. AB - OBJECTIVE: Abbe and Payr introduced vascular techniques and devices to facilitate vessel anastomosis over a century ago. Obora published the idea of a sutureless vascular anastomosis with use of magnetic rings in 1978. The purpose of this study was to assess the performance of a new magnetic device to perform a side-to side arteriovenous anastomosis in a dog model. MATERIAL AND METHODS: Male fox hounds (25 kg) were treated preoperatively and daily postoperatively with clopidogrel bisulfate (Plavix) and aspirin. The femoral artery and vein were exposed unilaterally in 3 dogs and bilaterally in 4 dogs (n = 11 anastomoses). A 4-mm arteriotomy was performed, and 1 oval magnet 0.5 mm thick was inserted into the lumen of the artery and a second magnet was applied external to the artery, compressing and stabilizing the arterial wall to create a magnetic port. An identical venous magnetic port was created with another pair of oval magnets. When the 2 ports were allowed to approach each other, they self-aligned and magnetically coupled to complete the arteriovenous anastomosis. Patency was assessed for the first hour with direct observation, again after 9 weeks with duplex ultrasound scanning, and at 10 weeks under direct open observation. The anastomoses were explanted after 10 weeks. Hydrodynamic resistance was measured ex vivo on the final 8 anastomoses by measuring the pressure drop across an anastomosis with a known flow rate. RESULTS: After implantation, very high flow created visible turbulence and palpable vibration. All 11 anastomoses were patent under direct observation and palpation. Ten of 11 anastomoses were clearly patent on duplex scans, and patency of 1 anastomosis was questionable. Hydrodynamic resistance averaged 0.73 +/- 0.33 mm Hg min/mL (mean +/- SEM). CONCLUSIONS: Vascular anastomoses performed with magnets demonstrated feasibility; exhibited 100% patency after 10 weeks in a dog arteriovenous shunt model; lacked apparent aneurysm or other potentially catastrophic failure; demonstrated remodeling of the vessel wall after several weeks to incorporate the magnets, making the magnetic force unnecessary; and warrants further study in vessels with different sizes, flow rates, and locations. CLINICAL RELEVANCE: We present a magnet-based device used to perform side-to-side peripheral vascular anastomoses. Its advantages include the ability to anastomose vessels without requiring circumferential surgical exposure. Vascular anastomosis performed with these magnets demonstrated 100% patency in the dog, lacked apparent aneurysm or other potentially catastrophic failure, and demonstrated remodeling of the vessel wall after several weeks, to incorporate the magnets, making indefinite retention of field strength unnecessary. This technique could enable minimally invasive procedures, such as complex reconstructive and revascularizing surgery, and warrants further study in vessels with different sizes, flow rates, and locations. PMID- 15337882 TI - Aortic wall cell proliferation via basic fibroblast growth factor gene transfer limits progression of experimental abdominal aortic aneurysm. AB - OBJECTIVE: Our previous study demonstrated that high flow conditions stimulated cell proliferation in the aortic wall in a rat model of abdominal aortic aneurysm (AAA), and we speculated that there is a possible relation between medial cell density and aortic wall integrity. In the present study we delivered the basic fibroblast growth factor (bFGF) gene to the aortic wall of a rat AAA model and evaluated the effects of growth factor-enhanced smooth muscle cell (SMC) proliferation on aneurysm progression. METHODS: AAA was induced in rats by means of infusion of porcine pancreatic elastase. Immediately after elastase infusion the abdominal aorta was filled with an expression plasmid vector containing the bFGF gene (bFGF group) or LacZ gene (control group); then gene transfer to the aortic wall was carried out with an in vivo electroporation method. The animals were killed 7 days after treatment, and the aneurysm was measured. The numbers of SMCs, macrophages, and endothelial cells were counted with immunostaining, and cell replication was evaluated with bromodeoxyuridine (BrdU) staining. RESULTS: Aneurysm diameter in the bFGF group was significantly smaller than that in the control group (4.6 +/- 0.3 mm vs 6.5 +/- 1.4 mm; P <.01). The numbers of medial SMCs and BrdU-incorporated cells in the bFGF group were significantly greater than those in the control group (SMC, 101 +/- 34 per high-power field [hpf] vs 80 +/- 31/hpf; P <.05, BrdU, 107 +/- 63/hpf vs 50 +/- 33/hpf; P <.05), whereas no difference was detected in the numbers of macrophages and endothelial cells between the 2 groups. CONCLUSIONS: Delivery of bFGF to the aortic wall induced significant enhancement of medial SMC proliferation, without an increase in inflammatory infiltration, then successfully limited aneurysm enlargement. These findings suggest that increased medial cellularity inhibits aneurysm formation, which possibly offers a clue for developing a new strategy for treatment of AAAs. PMID- 15337883 TI - Regulator of G protein signaling 5 marks peripheral arterial smooth muscle cells and is downregulated in atherosclerotic plaque. AB - OBJECTIVE: Regulator of G protein signaling 5 (RGS5), an inhibitor of Galpha(q) and Galpha(i) activation, was recently identified among genes highly expressed in smooth muscle cells (SMCs) of aorta but not vena cava. This finding prompted the hypothesis that RGS5 provides long-term G protein inhibition specific to normal arterial SMC populations and that loss of expression may in turn contribute to arterial disease. METHODS: To test this hypothesis we characterized RGS5 gene expression throughout the vasculature of nonhuman primates to determine whether RGS5 was restricted to arteries in other vascular beds and whether expression was altered in arterial disease. RESULTS: In situ hybridization localized RGS5 message to medial SMCs of peripheral arteries, including carotid, iliac, mammary, and renal arteries, but not accompanying veins. SMCs of many small arteries and arterioles also expressed RGS5, including glomerular afferent arterioles critical to blood pressure regulation. Differential expression persisted in culture, inasmuch as RGS5 message was significantly higher in SMCs derived from arteries than from veins at real-time polymerase chain reaction. It was remarkable that the only major arterial bed lacking RGS5 was the coronary circulation. In atherosclerotic peripheral arteries RGS5 was expressed in medial SMCs, but was sharply downregulated in plaque SMCs. CONCLUSION: These data identify RGS5 as a new member of a short list of genes uniquely expressed in peripheral arteries but not coronary arteries. Persistence of an arterial pattern of RGS5 expression in culture and lack of expression in coronary arteries support a unique SMC phenotype fixed by distinct lineage or differentiation pathways. The association between loss of expression and arterial wall disease has prompted the new hypothesis that prolonged inhibition by RGS5 of vasoactive or trophic G protein signaling is critical to normal peripheral artery function. PMID- 15337884 TI - Adventitial vasa vasorum heterogeneity among different vascular beds. AB - INTRODUCTION: Different vascular beds show substantial variation in their susceptibilities for development of vascular disease like atherosclerosis, and thereby exhibit a variety of different clinical presentations. Yet, the underlying mechanism of this heterogeneity is not well defined. Recent evidence suggests a role for the vasa vasorum (VV) in vascular disease. We hypothesized that there is a differential distribution structure of adventitial VV in different vascular beds. Hence, the current study was designed to characterize and compare the structure of the adventitial VV in the coronary and the peripheral circulation. METHODS: Samples of vessels from different vascular beds were obtained from 6 female crossbred domestic pigs. The samples were scanned using micro-computed tomography, and the images reconstructed and analyzed to characterize VV architecture, including vessel wall area, VV count, VV density, intravessel spatial distribution, mean diameter of first- and second-order VVs and the ratio of second- to first-order VVs. RESULTS: There were significant differences in VV density among different vascular beds. Density was highest in coronary arteries (2.91 +/- 0.26 vessels/mm2, P <.05, vs renal, carotid, and femoral arteries), intermediate in renal arteries (1.45+/- 0.22 vessels/mm2, P <.05, vs femoral artery) and carotid arteries (0.64 +/- 0.08 vessels/mm2, P <.05, vs femoral artery), and lowest in femoral arteries (0.23 +/- 0.05 vessels/mm2 ). A similar pattern for the ratio of second- to first-order VV was also observed. Random intravessel spatial distribution of VVs was seen in all vascular beds. CONCLUSION: The current study demonstrates a differential structure of the adventitial VV in different vascular beds. This intra- and intervessel heterogeneity in VV anatomy is a phenotypic variability that might determine a differential local response to systemic risk factors and, thereby, variable propensity for vascular disease among different vascular beds. PMID- 15337885 TI - Deep vein thrombosis resolution is not accelerated with increased neovascularization. AB - INTRODUCTION: Deep venous thrombosis (DVT) resolution involves fibrinolysis, neovascularization, and fibrosis. We hypothesized that promoting neovascularization would accelerate DVT resolution. METHODS: A rat model of stasis DVT was produced with proximal ligation of the inferior vena cava (IVC) and all visible tributaries. One microg of interferon inducible protein (IP-10; angiostatic chemokine), basic fibroblast growth factor (bFGF; pro-angiogenic cytokine), epithelial neutrophil activating protein (ENA-78; pro-angiogenic chemokine), or saline solution control was injected into the IVC after ligation, and then via tail vein injection daily until sacrifice at either 4 or 8 days. Peripheral blood counts were measured, and thrombus weight was recorded at sacrifice. Laser Doppler in vivo imaging was used to estimate post-thrombotic IVC blood flow. Immunohistologic assessment of the thrombosed IVC for polymorphonuclear neutrophils (PMNs), monocytes (ED-1), and laminin (neovascular channels) was performed or the thrombus was separated from the IVC and assayed for keratinocyte cytokine (KC), monocyte chemotactic protein-1 (MCP-1), bFGF with enzyme-linked immunosorbent assay (ELISA), and total collagen with a direct colorimetric assay. RESULTS: Peripheral blood and intrathrombus PMNs and monocytes were not significantly different in the treated or control rats. There were no differences in any measure at 4 days. At 8 days, thrombus neovascularity, but not weight or collagen content, was increased in rats treated with bFGF or ENA-78 compared with control rats (17.6 +/- 0.93, 16.2 +/- 0.97 vs 13.2 +/- 0.79; channels/5 high-power fields (hpf; n = 6-10; P <.05). Post DVT IVC blood flow was significantly increased in bFGF-treated rats but not in rats treated with IP-10 or ENA-78, as compared with control rats. Rats treated with ENA-78 had increased intrathrombus bFGF compared with control rats (85 +/- 27 pg/mg protein vs 20 +/- 6 pg/mg protein; n = 6; P <.05), but other mediators were not significantly different in treated rats compared with control rats. CONCLUSION: Pro-angiogenic compounds increase thrombus neovascularization, but this does not correlate with smaller or less fibrotic DVT. Mechanisms other than neovascularization may be more important to hasten DVT dissolution. Clinical relevance Improved therapy for deep venous thrombosis (DVT) will ideally increase the rate of thrombus dissolution and eliminate the bleeding risks of anticoagulation. This study evaluated promoting DVT neovascularization with angiogenic chemokines, and, while successful by experimental measures, this did not translate into smaller DVT. Solely promoting thrombus neovascularization will not likely speed resolution. PMID- 15337886 TI - Abdominal aortic aneurysm causing duodenal obstruction: two case reports and review of the literature. AB - Obstruction of the upper gastrointestinal tract caused by an abdominal aortic aneurysm (aortoduodenal syndrome) has been rarely reported. The typical presentation includes protracted emesis in a patient with a pulsatile abdominal mass. Clinical features of weight loss, abdominal pain, and distention are present less frequently. The diagnosis is suggested by findings on computed tomography scans, and may be confirmed with upper gastrointestinal contrast material-enhanced studies or upper endoscopy. Aortic aneurysmorrhaphy is curative, and should be undertaken after gastrointestinal decompression and correction of fluid and electrolyte disturbances. We report 2 cases of abdominal aortic aneurysm producing upper gastrointestinal obstruction, and provide a review of the literature relevant to this clinical syndrome. PMID- 15337887 TI - Intimal angiosarcoma of the aorta: report of a case and review of the literature. AB - Although extremely rare, the group of primary malignant tumors of the aorta (PMTA) exhibits enormous histologic heterogeneity. In most cases, diagnosis is established late in the course of the disease; the median survival time is only a few months. We present the case of a 75-year-old patient with an intimal angiosarcoma of the infrarenal abdominal aorta and discuss the clinical presentation, histopathologic diagnosis, and classification of primary aortic sarcomas. A critical review of the diagnostic and therapeutic management in this case revealed that the atypical aortic thrombus should have prompted a comprehensive preoperative diagnostic work-up, specifically with magnetic resonance tomography of the aorta and bone scintigraphy. PMID- 15337888 TI - Resection of an intravenous leiomyoma with intracardiac extension with use of endovascular techniques. AB - Intravenous leiomyoma with intracardiac extension is a rare entity that necessitates surgical excision to alleviate symptoms and prevent life-threatening complications. These procedures are generally performed under cardiopulmonary bypass, and review of the literature reveals an evolution in management of this disease from 2-stage to single-stage procedures. We report the case of a 45-year old woman with an intravenous leiomyoma with intracardiac extension after presenting with syncopal symptoms. The intravenous tumor and its supradiaphragmatic extension were ultimately resected via laparotomy without the need for cardiopulmonary bypass. Distal control was achieved by passing an occluding balloon catheter above the tumor through a venotomy with fluoroscopic guidance and echocardiography, thus enabling protected tumor extraction through the intra-abdominal inferior vena cava. In selected patients, we believe use of endovascular techniques combined with proper preoperative imaging can safely permit single-stage resection of these tumors via laparotomy without cardiopulmonary bypass. PMID- 15337889 TI - Use of superficial femoral artery to treat an infected great vessel prosthetic graft. AB - We report a patient treated for infection of an ascending aorta to bilateral common carotid artery bypass graft. The superficial femoral arteries were used for the reconstruction after local treatment failed. The patient is free from infection and the grafts are patent 4 years after operation. We believe this is the only report in the literature in which the superficial femoral arteries were used to reconstruct an infected great vessel graft. PMID- 15337891 TI - Chronic posttraumatic aneurysm of descending aorta with fistulous communication into pulmonary artery. AB - We report an unusual case of presumed posttraumatic chronic proximal descending aneurysm associated with a fistula into the pulmonary artery. Review of the literature showed that a fistula of the descending aorta into the pulmonary artery is uncommon and that only a few patients have survived. This rare entity is best treated with open repair; however, in patients at high risk an endovascular stent graft is an option. PMID- 15337892 TI - Repair of mycotic aortic pseudoaneurysm with a stent graft using transesophageal echocardiography. AB - A 54-year-old man who underwent uneventful orthotopic heart transplantation 1 year previously had low-grade fever and dyspnea. Imaging studies revealed an ascending aortic pseudoaneurysm (AAP), which was repaired with a 5-mm polyester patch, with circulatory arrest and cardiopulmonary bypass. Intraoperative cultures of the AAP grew methicillin-resistant Staphylococcus aureus, and the pseudoaneurysm recurred after 6 weeks despite intravenously administered antibiotic therapy. A 28.5-mm x 3.3-cm Gore Excluder aortic cuff was deployed in the ascending aorta through a left axillary artery cutdown with use of combined transesophageal echocardiography and fluoroscopy. In addition, controlled hypotension and asystole were established with administration of adenosine to facilitate precise device deployment. Postoperative imaging with transesophageal echocardiography and magnetic resonance angiography revealed complete resolution of the AAP, and the patient had done well at 7-month follow-up. Treatment of a mycotic aortic pseudoaneurysm with an endoprosthesis in a patient without other treatment alternatives can be performed safely, with acceptable short-term results. PMID- 15337893 TI - Embolus of surgical adhesive to the extremities causing acute ischemia: report of two cases. AB - We present 2 cases in which a surgical adhesive embolized to the extremities. In the first case an adhesive was successfully used in the repair of a DeBakey type I aortic dissection. The patient was seen 2 months postoperatively with acute lower extremity ischemia, and a large piece of adhesive was extracted from the iliac and femoral arteries at embolectomy. In the second case the adhesive was used to seal a pericardial patch during repair of a ventricular septal defect. This patient was seen 1 day postoperatively with acute arm ischemia, and the adhesive particle was extracted from the brachial artery during embolectomy. PMID- 15337894 TI - Occlusion of a giant splenic artery pseudoaneurysm with percutaneous thrombin collagen injection. AB - Splenic artery pseudoaneurysms have been conventionally treated with surgical or endovascular techniques, including transcatheter embolization. This case report describes the use of procoagulant components thrombin and collagen in large volume to percutaneously embolize a giant splenic artery pseudoaneurysm. PMID- 15337895 TI - The Giacomini vein as an autologous conduit in infrainguinal arterial reconstruction. AB - The standard conduit in infrainguinal arterial bypass grafting, the great saphenous vein, is often unavailable. Arm and small saphenous veins are used as alternative conduits; yet both are deficient in length to accommodate femorocrural bypasses as a single conduit. In light of its high prevalence, the Giacomini vein harvested in continuity with the small saphenous vein may offer the latter extra length, promoting their combination into a single conduit able to meet the needs of infrainguinal reconstruction, particularly in lengthy infrainguinal bypass grafting. The Giacomini vein merits consideration when arterial reconstruction is performed in proximity to its anatomic course. PMID- 15337896 TI - Ruptured abdominal aortic aneurysm with aorto-caval fistula. PMID- 15337897 TI - A live dog is better than a dead lion. PMID- 15337898 TI - Stem cell research: obligations when religious values conflict with professional values. PMID- 15337899 TI - Misperceptions regarding the long-term safety of the AneuRx stent graft. PMID- 15337900 TI - Inappropriate industry influence on FDA processes. PMID- 15337901 TI - Regarding "Withdrawal of article by the FDA after objection from Medtronic". PMID- 15337902 TI - Regarding "Carotid endarterectomy in SAPPHIRE-eligible high-risk patients: implications for selecting patients for carotid angioplasty and stenting". PMID- 15337914 TI - Advocacy for children through activism. PMID- 15337915 TI - The limping child. AB - A child who limps often presents a diagnostic challenge. The differential diagnosis is extensive. Although the most common cause is trauma, awareness of other potential causes is important. The age of the child and the pattern of the gait help narrow the differential diagnosis. In most cases, a diagnosis can be made from the history and physical examination. If the diagnosis is not obvious after a careful clinical evaluation, plain radiographs provide an excellent means of screening for fracture, joint effusion, lytic lesions, periosteal reaction, and avascular necrosis. Other tests should only be ordered when indicated. PMID- 15337916 TI - Rate of palivizumab administration in accordance with current recommendations among hospitalized children. AB - INTRODUCTION: The American Academy of Pediatrics recommends palivizumab prophylaxis for children born premature or with chronic lung disease to reduce the severity of respiratory syncytial virus (RSV) infection. The purpose of the current study is to examine palivizumab administration among children hospitalized with RSV infection. METHOD: This is a retrospective medical record review at a tertiary care children's hospital. The study included children <2 years of age hospitalized between October 1, 2000, and April 1, 2001, with the diagnosis of RSV infection. RESULTS: The records of 264 children meeting inclusion criteria were reviewed. Forty children qualified for administration of palivizumab using American Academy of Pediatrics recommendations. Of these, 14 (35%) received palivizumab prior to admission. Palivizumab administration rate was not affected by age, race, or insurance coverage. DISCUSSION: Exact barriers to the administration of palivizumab remain unclear. The identification of high risk children, prevention of RSV by use of palivizumab, and collaboration between hospital and community health care providers will help increase the use of palivizumab and decrease the incidence of RSV. PMID- 15337917 TI - Parents' attitudes to children's tobacco smoke exposure and how the issue is handled in health care. AB - INTRODUCTION: The objective of the study was to understand the opinions and attitudes among parents of preschool children towards children's passive smoking, to show how attitudes influenced smoking and smoking behavior, and how the parents had experienced the handling of the tobacco issue in antenatal and child health care. METHOD: A subsample of smoking and nonsmoking parents (n = 300) with 4- to 6-year-old children participating in All Babies in Southeast Sweden (ABIS), a prospective study on environmental factors affecting development of immune mediated diseases, answered a questionnaire on their opinions and attitudes to children's passive smoking. RESULTS: Indoor smokers were more positive regarding smoking, less aware of the adverse health effects from passive smoking, and more negative regarding the handling of tobacco prevention in health care than both outdoor smokers and nonsmokers. Indoor smokers' idea of how children should be protected from tobacco smoke exposure was significantly different from the idea of nonsmokers and outdoor smokers. DISCUSSION: Results indicate that further intense efforts are needed to convince the remaining indoor smokers about the adverse health effects related to tobacco smoke exposure. Pediatric nurses meet these parents in their daily work and should be aware of the need to focus this group and their use of protective measures. PMID- 15337918 TI - Breaking the cycle of family violence: understanding the perceptions of battered women. AB - BACKGROUND: Domestic violence screening recommendations that guide pediatric care providers exist but screening remains low. Many providers do not understand the linkages between aversive childhood experiences and adult abuse experiences. PURPOSE: To describe the childhood abuse experiences in 40 battered women. Research questions How prevalent is childhood abuse in a sample of battered women? How do these women describe their childhood abuse experiences? METHOD: Women who had left or were currently in abusive relationships were recruited by posting advertisements in public areas. Each woman participated in a semi structured individual interview that included open-ended questions and administration of standardized measures. RESULTS: Most women described abuse during their childhood, reported high depressive symptoms, and indicated a high potential risk for child abuse. Content analysis of descriptions of childhood abuse experiences yielded five themes: socialization not to tell, normalcy of events, abandonment, escape, and shame. Women reported limited assistance during their childhood and adult abuse experiences from health care providers who lacked a family focus. DISCUSSION: Mothers' experiences with childhood and adult abuse challenge their ability to develop healthy family relationships. Pediatric health care providers must routinely screen for abuse in mothers and children within a family-centered approach. PMID- 15337919 TI - Father stress during a child's critical care hospitalization. AB - INTRODUCTION: There is a dearth of research on the experiences of fathers with critically ill children. The specific aim of this study was to examine stress and specific stress symptoms of fathers with a child in the pediatric intensive care unit (PICU) and compare their stress and stress symptoms with those of fathers with a child in a general care unit (GCU). METHOD: In this descriptive, comparative study, fathers were recruited from a large Midwestern children's hospital within 24 hours of their child's admission. The sample included 15 PICU and 10 GCU fathers who completed the Parental Stressor Scale: PICU and the Symptom Checklist-90-Revised. RESULTS: PICU fathers perceived significantly greater stress than did GCU fathers from the sights and sounds of the unit and by procedures their child underwent. PICU fathers also reported greater frequencies of stress symptoms, such as headache, low energy, annoyance, and worrying. DISCUSSION: Advanced practice and staff nurses should know how fathers of critically ill children feel and what stresses them so they can provide interventions specific for fathers during this stressful time. PMID- 15337920 TI - Contraception for adolescents: Part two. PMID- 15337921 TI - Jessica's summer growth spurt. PMID- 15337922 TI - Patient education. Using nonpharmacological treatments in conjunction with stimulant medications for children with ADHD. PMID- 15337925 TI - Use of health diaries in pediatric research. PMID- 15337926 TI - School backpacks: preventing injuries. PMID- 15337927 TI - Position statement: Reimbursement for nurse practitioner services. PMID- 15337929 TI - Wire strikes--the hidden menace. Part I. PMID- 15337930 TI - Exogenous surfactant administration during the transport of premature infants. PMID- 15337932 TI - Critical care education matrix revisited. PMID- 15337933 TI - Voices of our past: flight nurse training in World War II. PMID- 15337934 TI - University Air Care marks 2 decades in Cincinnati. PMID- 15337954 TI - A retrospective analysis of the intubations performed during 5 years of helicopter emergency medical service in Amsterdam. AB - INTRODUCTION: Different skilled personnel perform prehospital airway management, by far one of the most challenging skills with major consequences upon failure. SETTING: The setting for this study was the helicopter emergency medical service at the Vrije Universiteit Medical Center, Amsterdam, the Netherlands. METHODS: We conducted a retrospective analysis of all medical charts of intubated trauma patients in the period from May 1995 to May 2000. We focused on intubation reasons and conditions. RESULTS: In 43 of 653 patients (7%) the process of intubation was recorded as being difficult, leading to 5 failed intubations (11.6%). In 432 of 653 trauma victims (66%), general anaesthesia was required before intubation. Forty (9%) of these patients died, most soon after arrival in the hospital. The clinical condition of 221 (34%) patients was so poor that they did not require additional drugs for intubation; 73% of those patients died, with two-thirds dying at the accident site. CONCLUSION: The rate of difficult intubation in this analysis is low (7%). The overall airway failure (11.6%) is the same as seen in the literature when sedation and relaxation are used. An adult trauma victim with a Revised Trauma Score of 0 has a very poor prognosis of survival. PMID- 15337955 TI - EMS certification requirements for flight nurses. AB - INTRODUCTION: Emergency medical technician (EMT) or paramedic (EMTP) certification requirements for flight nurses (FNs) providing on-scene patient care vary. We surveyed those requirements and evaluated the relationships between flight team composition or program location and FN EMS certification. METHODS: Telephone survey of all 184 rotor-wing programs responding with a nurse to scenes RESULTS: The overall EMS training requirement for FNs was: none-57.6%, EMT-21.7%, EMTP-14.7%, local credential (not EMT or EMTP)-6.0%. Second team members were EMTP, RN, physician, or respiratory therapist (RRT). Overall, team configuration related significantly to FN EMS certification (P =.01). FN/EMTP and FN/RRT teams were individually significant (P <.01), with FN/EMTP teams tending not to require certification and all FN/RRT teams tending toward a certification requirement. Neither FN/FN nor FN/physician pairings related significantly with FN EMS certification requirements. Regional patterns emerged to both crew configuration and FN EMS certification requirements. CONCLUSION: Most flight programs do not require FN EMT/EMTP certification. Team configuration and geography are related to those requirements. PMID- 15337956 TI - Value of myocardial contrast echocardiography for predicting left ventricular remodeling and segmental functional recovery after anterior wall acute myocardial infarction. AB - OBJECTIVE: We sought to study the value of microvascular perfusion assessed by myocardial contrast echocardiography in predicting left ventricular remodeling after anterior wall acute myocardial infarction. METHODS: In 31 patients myocardial contrast echocardiography was performed up to 48 hours after acute myocardial infarction with determination of end-diastolic and end-systolic volumes, wall-motion score index, and myocardial perfusion score index (MPSI) at rest and under dobutamine stress at 6 months. Patients were classified into remodeling group (RG) (n = 19) and non-RG (n = 12), and, according to number of segments without opacification, reflow (< or =2 segments, n = 15) and no-reflow (>2 segments, n = 16) groups. RESULTS: Wall-motion score index (1.84 +/- 0.22 vs 1.64 +/- 0.3; P =.049), MPSI (1.53 +/- 0.25 vs 1.26 +/- 0.17; P =.006), and number of segments without contrast (3.11 +/- 2.23 vs 1.08 +/- 1.38; P =.018) were higher in RG than in non-RG. End-diastolic and end-systolic volumes, and wall-motion score index, increased significantly in RG at 6 months and decreased in non-RG. MPSI increased in RG (1.53 +/- 0.25-1.66 +/- 0.21; P =.011) and was the only independent predictor of left ventricular remodeling (odds ratio = 1.8; 95% confidence interval = 1.15-2.82; P =.010). No-reflow group presented 27.8 +/- 19.9% of segments with resting functional recovery or contractile reserve, and reflow group presented 69.9 +/- 31.2% (P <.001). CONCLUSION: MPSI obtained 48 hours after acute myocardial infarction is an independent predictor of left ventricular remodeling. Patients with two or fewer segments without opacification revealed a better prognosis of resting ventricular function and contractile reserve. PMID- 15337957 TI - Diagnosis of viable myocardium using velocity data of Doppler myocardial imaging: comparison with positron emission tomography. AB - To test whether velocity data of Doppler myocardial imaging (DMI) at rest is useful for diagnosis of myocardial viability, 25 consecutive patients (age 64 +/- 10 years) with regional wall-motion abnormalities at the left anterior descending coronary artery territory and left ventricular dysfunction (ejection fraction: 31 +/- 7%) underwent both DMI at rest and positron emission tomography. The peak systolic velocity (Vpeak) and postsystolic thickening (PST) velocity were measured in myocardial segments of left anterior descending coronary artery territory from apical views. A total of 71 segments were classified by positron emission tomography as normal or viable in 38 (group A) and nonviable in 33 (group B). Although Vpeak did not show any difference between groups (1.81 +/- 1.77 vs 1.29 +/- 0.94 cm/s, P =.107), PST velocity was significantly higher in group A (2.48 +/- 1.68 vs 0.89 +/- 0.72 cm/s, P <.001). The sensitivity and specificity of PST velocity > 2.0 cm/s for diagnosis of viability were 61% (23/38) and 97% (32/33), respectively. In segments with PST velocity was < or =2.0 cm/s, Vpeak > 1.8 cm/s could discriminate group A from B with a sensitivity of 67% (10/15) and a specificity of 91% (29/32). The algorithm using both PST velocity and Vpeak of DMI showed sensitivity and specificity of 87% and 88%, respectively, for diagnosis of myocardial viability. Velocity data of DMI at rest provides robust information regarding viability in selected patients, and an advantage of this technique is that no stress testing is needed. PMID- 15337958 TI - Influence of preload alterations on parameters of systolic left ventricular long axis function: a Doppler tissue study. AB - The study examined the influence of preload alterations on parameters of left ventricular (LV) long-axis function during systole. Doppler tissue echocardiography was performed in 32 healthy volunteers at rest who were placed in the Trendelenburg position after administration of 0.8 mg of nitroglycerin sublingually. In a 16-segment LV model, long-axis myocardial strain and systolic velocities were assessed. Isovolumic acceleration was assessed in the lateral mitral annulus. Mean LV strain and systolic velocity were unchanged during the different load conditions. In the mitral annulus, strain was also uninfluenced by preload alterations, whereas the peak systolic velocities seemed influenced by preload reduction. The isovolumic acceleration of the mitral annulus was found to be load dependent (enhanced preload 1.38 +/- 0.50 vs baseline 1.6 m/s2 +/- 0.60, P <.01; and preload reduction 2.18 +/- 0.65 m/s2, P <.01). In conclusion, LV long axis strain and systolic velocities were not significantly influenced by load alterations, whereas myocardial systolic velocities and isovolumic acceleration in the lateral mitral annulus were significantly load dependent. PMID- 15337959 TI - Relationship of systolic dysfunction to area at risk and infarction size after ischemia-reperfusion in mice. AB - Whereas the extent of wall-motion abnormality (WMA) correlates well with the area at risk (AAR) and infarction size in murine models of nonreperfused myocardial infarction, this relationship is less clear in the setting of ischemia reperfusion injury. Echocardiography was performed in mice at baseline, after left anterior descending coronary artery ligation (30 minutes) followed by reperfusion (24 hours), and after religation of the left anterior descending coronary artery. The extent of WMA before and after religation was compared with the initial infarction size measured by triphenyltetrazolium chloride and the AAR measured by fluorescent microspheres, respectively. Echocardiography showed left ventricular dilation and dysfunction after ischemia and reperfusion. WMA after religation correlated well with AAR (r2 = 0.70, P <.0001). The correlation between WMA and infarction size was incomplete (r2 = 0.59, P <.0002) in part because of underestimation of nontransmural infarcts. Echocardiography can reliably assess AAR after ischemia-reperfusion in mice; however, it does not allow for precise quantification of the small areas of necrosis that often occur in this setting. PMID- 15337960 TI - Transthoracic Doppler validation in mammary artery grafts after minimal invasive direct coronary artery bypass operation. AB - OBJECTIVE: This study was performed to validate noninvasive transthoracic Doppler ultrasound (TTD) with simultaneous invasive Doppler guidewire measurements in patients after minimal invasive direct coronary artery bypass operation. METHODS: A total of 14 patients were examined 3 to 8 days after minimal invasive direct coronary artery bypass operation. TTD was performed to measure systolic and diastolic peak velocities of the left internal mammary artery (LIMA) at rest and during adenosine-induced hyperemia. Simultaneous Doppler guidewire measurements were performed. RESULTS: LIMA flow was detected in 12 of 14 patients (86%). There was high agreement between TTD and Doppler guidewire measurements of LIMA flow velocities (systolic peak velocity: r = 0.86, y = 11.3 + 0.82x +/- 7.9; diastolic peak velocity: r = 0.95, y = 5.7 + 1.02x +/- 7.5; average peak velocity: r = 0.95, y = 5.2 + 0.94x +/- 5.4; and flow velocity reserve: r = 0.97, y = 5.2 + 0.99x +/- 4.5). CONCLUSION: TTD represents an accurate method to evaluate flow velocities and flow velocity reserve of LIMA bypass grafts even in the early phase after minimal invasive direct coronary artery bypass operation. PMID- 15337961 TI - Anatomic M-mode, a pertinent tool for the daily practice of transthoracic echocardiography. AB - OBJECTIVES: We sought to compare anatomic M-mode (AMM), a new echocardiographic postprocessing option, and conventional M-mode (CMM) using fundamental imaging and tissue harmonic imaging. METHODS: Transthoracic echocardiography was performed in 15 selected patients to analyze the reproducibility of AMM and in 47 patients to assess its clinical value versus CMM. Acquisitions were performed successively: CMM fundamental imaging; CMM tissue harmonic imaging; tissue harmonic imaging cineloops for AMM; and fundamental imaging cineloops for AMM. Quantitative analysis was performed offline. The angle alpha between the CMM line and the septal endocardial interface was calculated and the expected percentage of error in measuring left ventricular diameter was derived. RESULTS: AMM analysis was reproducible. Optimal AMM full echocardiographic definition was obtainable in 77% of the population, whereas CMM was optimal for 49% because of scan line misalignment, causing a measurement overestimation exceeding 5%. CONCLUSION: The ability with AMM to reduce the alpha angle to 0 degrees and, thus, avoid overestimation of left ventricular dimensions might improve follow-up in several pathologic conditions. PMID- 15337962 TI - Determinant of left atrial dilation in patients with hypertrophic cardiomyopathy: a real-time 3-dimensional echocardiographic study. AB - To identify the determinants of left atrial (LA) dilation for patients with hypertrophic obstructive cardiomyopathy (HOCM), first we validated LA volume determination by real-time 3-dimensional echocardiography using magnetic resonance imaging in patients. Subsequently, real-time 3-dimensional echocardiography and 2-dimensional Doppler echocardiography were performed in 60 patients with HOCM and in 17 age-matched control subjects. LA volumes and left ventricular (LV) filling pressures were higher for patients with HOCM than in control subjects. By stepwise multilinear regression analysis, LV end-diastolic pressure, resting LV outflow tract pressure gradient, and LV wall thickness were significant determinants of LA dilation. However, tau, -dP/dt, LV stiffness, provokable pressure gradient, and mitral regurgitation did not have any independent relationship with LA volumes. Therefore, it is concluded that LA volume can be accurately determined by real-time 3-dimensional echocardiography; and LA dilation in patients with HOCM is related to LV filling pressure, LV outflow tract pressure gradient, and LV wall thickness. PMID- 15337963 TI - Comparison of real-time 3-dimensional echocardiography with conventional 2 dimensional echocardiography in the assessment of structural heart disease. AB - We evaluated the diagnostic use of a real-time 3-dimensional (3D) echocardiographic system in 106 patients referred for echocardiography during a 4 month period. Real-time 3D echocardiography was performed and recorded in parallel with a routine, comprehensive 2-dimensional (2D) study. The diagnoses were exclusively on the basis of 2D findings. The 3D volumes were sliced offline in the 3 dimensions to selectively display specific cardiac structures and reviewed independent of the 2D findings. The 3D studies were graded as: A, new finding not on 2D studies; B, useful anatomic perspective; C, equivalent to 2D studies; or D, missed 2D findings. Compared with 2D echocardiography, 3D echocardiography was graded A in 7 (7%), B in 19 (18%), C in 65 (61%), and D in 15 (14%) cases. In the 26 grade-A and grade-B studies, mitral valve disease and congenital heart disease accounted for 16 (61%) cases. Suboptimal image quality was present in 7 (47%) of the 15 grade-D studies. Thus, real-time 3D echocardiography yields anatomic information comparable with conventional 2D echocardiography in the majority of patients. It can provide new and useful anatomic insight, particularly in patients with mitral valve disease and congenital heart disease. Suboptimal image quality remains a problem for real time 3D echocardiography in some patients. PMID- 15337964 TI - In vivo short-term Doppler hemodynamic profiles of 189 Carpentier-Edwards Perimount pericardial bioprosthetic valves in the mitral position. AB - We sought to determine the hemodynamic performance of the Carpentier-Edwards Perimount pericardial valve in the mitral position. We reviewed the Doppler echocardiographic data on 189 patients (110 women; 68 +/- 12 years of age) who were implanted with this valve (7.6 days +/- 13 postoperatively) at our institution between September 2000 and May 2002. The average ejection fraction was 47%. For all valves, the peak velocity was 1.9 +/- 0.3 m/s, peak gradient was 15 +/- 4.8 mm Hg, and mean gradient was 5.8 +/- 2 mm Hg. The pressure half-time was 93 +/- 24 milliseconds, with a calculated effective orifice area of 2.5 +/- 0.6 cm2. The average effective orifice area by continuity equation (83 valves) was 1.5 +/- 0.5 cm2. The mitral regurgitation was graded mild or less in 97.5% of all valves. This is the largest series establishing the favorable hemodynamic behavior of the different sizes of a new Perimount mitral valve, and the reported data could serve as a reference. PMID- 15337965 TI - Identifying patients with degenerative mitral regurgitation for mitral valve repair and replacement: a transesophageal echocardiographic study. AB - OBJECTIVE: We sought to preoperatively identify the suitability of patients with degenerative mitral valve (MV) regurgitation for MV repair (MVR) and MV replacement. BACKGROUND: MVR is the preferred method of treatment over MV replacement, if surgically feasible. MVR preserves left ventricular function and decreases risk of hemolysis, thromboembolism, and-in the absence of anticoagulation-hemorrhage. However, the ability to identify patients suitable for MVR preoperatively is somewhat limited. METHODS: In all, 76 patients underwent MV operation for severe symptomatic mitral regurgitation. The decision to operate was at the discretion of the referring physician in consultation with respective cardiothoracic surgeons at two separate, nonrelated institutions. All patients underwent preoperative and/or intraoperative transesophageal echocardiographic studies. RESULTS: In all, 35 patients (46%) underwent MVR and 41 (54%) underwent MV replacement. There was no difference in the percentage of MVRs between the two institutions: 17 cases (41%) at Hahnemann University Hospital, Philadelphia, Pa, versus 18 cases (53%) at Northwestern University Memorial Hospital, Chicago, Ill (P = not significant). Age was found to be a significant univariate predictor with older age favoring MV replacement. On average, patients who underwent MVR were 11 years younger then those who underwent MV replacement. Heart failure was also found to be a significant univariate predictor: as New York Heart Association functional class worsened, MV replacement was more likely. Echocardiographic variables favoring MVR included chordal length (>29 mm, P <.001), length of posterior mitral leaflet (>17 mm, P <.008), and length of anterior leaflet (>25 mm, P <.01). The only echocardiographic parameter favoring replacement was the presence of anterior mitral annular calcification. Using multivariate analysis, older age (>63 years) was again a significant predictor favoring MV replacement (P <.002; odds ratio [OR] 20). Longer chordal length (>29 mm) was the strongest predictor favoring MVR (P <.001; OR 11.2). Longer length of the posterior leaflet (>17 mm; OR 5.07) and mitral annulus size > 35 mm (OR 7.75) were also significant multivariate predictors favoring MVR. The presence of anterior mitral annular calcification favored MV replacement using multivariate analysis (OR 25). CONCLUSIONS: Patients suitable for MVR can be identified preoperatively using a combination of clinical and echocardiographic parameters. PMID- 15337966 TI - Infective endocarditis resulting in rupture of sinus of Valsalva with a rupture site communicating with both the right atrium and right ventricle. AB - This report describes a patient with a rupture of the noncoronary sinus of Valsalva caused by erosion of its wall by a large right atrial vegetation not attached to the tricuspid valve without aortic valve involvement. The rupture site was located right posterior to the tricuspid septal leaflet and communicated both with the right atrium and the right ventricular inflow tract, resulting in a significant left-to-right shunt leading to severe right heart failure and shock. The diagnosis was made by transthoracic and transesophageal echocardiography. To our knowledge no similar case has been reported in the literature. PMID- 15337967 TI - Effect of a kink in unilateral pulmonary artery anastomosis on velocities of blood flow through bilateral pulmonary vein anastomoses in living-donor lobar lung transplantation. AB - Intraoperative transesophageal echocardiography is generally performed to detect vascular complications during lung transplantation. We report a case with a kink in pulmonary artery (PA) anastomosis suggested by an abnormal flow profile of pulmonary vein (PV) anastomoses during living-donor lobar lung transplantation. During the transplantation, velocity of blood flow through the right PV anastomosis showed abnormal elevation. Then, the patient's PA pressure elevated abnormally and a kink in the left PA anastomosis was found. Careful monitoring of PV anastomoses may enable detection of not only an abnormality of PV anastomoses but also that of PA anastomoses, especially in living-donor lobar lung transplantation. PMID- 15337968 TI - Cardiac granulocytic sarcoma (chloroma): in vivo diagnosis with transesophageal echocardiography. AB - Granulocytic sarcoma, or chloroma, is an uncommon presentation of acute leukemia. Cardiac involvement is very rare and usually diagnosed at autopsy. We present a case that discloses the essential role of transesophageal echocardiography for its in vivo diagnosis. The principal features with this imaging technique are finely described. PMID- 15337969 TI - Differences between flow profiles of pulmonary vein anastomoses affected by peripheral atelectasis in cadaveric and bilateral living-donor lobar lung transplantations. AB - We report two cases of peripheral atelectasis during cadaveric and living-donor lobar lung transplantation, which had different effects on the flow profile of pulmonary vein (PV) anastomoses. In the patient who underwent living-donor lobar lung transplantation, we detected the increase in the velocity of blood flow through the left PV anastomosis by intraoperative transesophageal echocardiography. Then peripheral atelectasis occurred in the transplanted left lung lobe. On the other hand, in the patient who underwent cadaveric bilateral lung transplantation, peripheral atelectasis occurred, but no changes in velocities of blood flow through PV anastomoses were detected by intraoperative transesophageal echocardiography. This difference may have been caused by the difference in sizes of pulmonary beds of transplanted grafts. These findings indicate the necessity of careful monitoring of PV anastomoses, especially in cases of living-donor lobar lung transplantation. PMID- 15337970 TI - Diagnosis of pulmonary embolism by acute right heart morphologic and hemodynamic changes observed during exercise stress echocardiography. AB - In addition to the conventional means of diagnosing pulmonary embolism, resting echocardiography has sometimes been useful. We describe the case of a patient with a normal resting transthoracic echocardiogram, but with a markedly abnormal posttreadmill exercise echocardiogram revealing acute right ventricular dilatation, marked ventricular septal shift, and acute pulmonary hypertension. Pulmonary embolism was suspected and subsequently confirmed by chest computed tomographic angiography. PMID- 15337971 TI - Left ventricular hydatid cyst presenting with acute ischemic stroke: case report. AB - Cardiac hydatid cysts are rarely seen. The presentation of an acute stroke secondary to embolization from a cardiac hydatid cyst is also rare. We report a young boy with left ventricular hydatid cyst who presented with acute ischemic stroke. PMID- 15337972 TI - Strain rate and strain: a step-by-step approach to image and data acquisition. AB - The advent of strain and strain rate imaging in clinical echocardiography prompted an examination of proper techniques for image acquisition and analysis. For this promising and relatively new method of assessing myocardial performance to be successful in diagnostic echocardiography, close adherence to a standardized method is imperative. This article provides the echocardiographer with a step-by-step approach. PMID- 15337973 TI - Rosacea: I. Etiology, pathogenesis, and subtype classification. AB - Rosacea is one of the most common conditions dermatologists treat. Rosacea is most often characterized by transient or persistent central facial erythema, visible blood vessels, and often papules and pustules. Based on patterns of physical findings, rosacea can be classified into 4 broad subtypes: erythematotelangiectatic, papulopustular, phymatous, and ocular. The cause of rosacea remains somewhat of a mystery. Several hypotheses have been documented in the literature and include potential roles for vascular abnormalities, dermal matrix degeneration, environmental factors, and microorganisms such as Demodex folliculorum and Helicobacter pylori. This article reviews the current literature on rosacea with emphasis placed on the new classification system and the main pathogenic theories. Learning objective At the conclusion of this learning activity, participants should be acquainted with rosacea's defining characteristics, the new subtype classification system, and the main theories on pathogenesis. PMID- 15337974 TI - Local neural block at the wrist for treatment of palmar hyperhidrosis with botulinum toxin: technical improvements. AB - BACKGROUND: Wrist blockage of median and ulnar nerves before treatment of palmar hyperhidrosis with botulinum toxin (BTX-A) reduces discomfort and improves accuracy of BTX-A injections, but can be associated with mechanical/chemical injury. OBJECTIVES: We sought to compare locoregional anesthesia of median and ulnar nerves using conventional 25-G x 0.50 x 13 mm gauge needle with short 30-G x 0.40 x 6 mm gauge needle. METHODS: In all, 37 patients with idiopathic, recalcitrant palmar hyperhidrosis were treated with BTX-A after median and ulnar nerve blockage. In 18 patients, a conventional needle was used to achieve nerve blockage and in 19 the short needle was used. The 2 groups of patients were compared for analgesic effects and lag phase. RESULTS: No differences were found between groups for lag phase (P=.26) and discomfort of subsequent BTX-A treatment (P=1.0). CONCLUSION: The use of a short-gauge needle to block median and ulnar nerves is a suitable method to anesthetize the palm before treatment with BTX-A. PMID- 15337975 TI - Prevalence and relevance of contact dermatitis allergens: a meta-analysis of 15 years of published T.R.U.E. test data. AB - BACKGROUND: The patch test procedure is frequently employed to help determine or confirm the cause of allergic contact dermatitis (ACD). The T.R.U.E. Test has become a global standard and is the commercially available patch test system currently used within the United States. Although many studies report T.R.U.E. Test data, none has measured the overall prevalence and relevance of reactions to the allergens tested by the T.R.U.E. Test. Our objective is to describe the prevalence and relevance of contact dermatitis allergens as tested by the T.R.U.E. Test. METHODS: We conducted a search of the MEDLINE database from 1966 to June 2000 for all publications on the use of the T.R.U.E. Test in the clinical evaluation of ACD in human subjects. Inclusion and exclusion criteria were applied. For each study, we identified and recorded the number of subjects tested, the number of patients with positive reactions, and the number with relevant reactions. Data were analyzed using the SAS system (Cary, NC). RESULTS: Ours is the first study to compile the entire corpus of published T.R.U.E. Test data and to examine these data using meta-analytic techniques. The meta-analysis shows that nickel (14.7% of tested patients), thimerosal (5.0%), cobalt (4.8%), fragrance mix (3.4%), and balsam of Peru (3.0%) are the most prevalent allergens. The 5 least prevalent allergens are paraben mix (0.5%), black rubber mix (0.6%), quaternium-15 (0.6%), quinoline mix (0.7%), and caine mix (0.7%). By contrast, North American Contact Dermatitis Data Group (NACDG) data show that the 5 most prevalent allergens are nickel (14.3%), fragrance mix (14%), neomycin (11.6%), balsam of Peru (10.4%), and thimerosal (10.4%). NACDG data indicate that the prevalence of allergy to cobalt is 9.2%. In order to assess the clinical importance of T.R.U.E. Test allergens, we employ the Significance-Prevalence Index Number (SPIN). Based on SPIN, the most clinically important allergens tested by the T.R.U.E. Test are nickel (SPIN=894), cobalt (266), fragrance mix (158), colophony (141), and thiuram mix (138). CONCLUSIONS: Our results identify the prevalence of common contact dermatitis allergens as tested by the T.R.U.E. Test and are in general agreement with previously published reports using other patch test methods. Over 3700 allergens have been identified as causing ACD, of which the T.R.U.E. Test tests only 23. Thus, the T.R.U.E. Test is a screening test at best. Comparison with NACDG data suggests that clinically important allergens may be missed by the T.R.U.E. Test. PMID- 15337976 TI - Halo nevus, rather than vitiligo, is a typical dermatologic finding of turner's syndrome: clinical, genetic, and immunogenetic study in 72 patients. AB - Turner's syndrome (TS) is a genetic disorder caused by numeric and/or structural abnormalities of the X chromosome; it is characterized by short stature, gonadal dysgenesis, and frequently by webbed neck, cubitus valgus, and lymphedema at birth. TS has been associated with several cutaneous abnormalities including an increased frequency of pigmented nevi, but few reports consider nevi in detail. Halo nevus (HN) is clinically defined as a melanocytic nevus surrounded by a halo of depigmentation. Vitiligo, a dermatologic disorder characterized by the presence of depigmented patches on the skin, has been described in the list of cutaneous findings associated with TS. The aim of this study was to determine the prevalence of HN and vitiligo in TS and to evaluate if a correlation between major histocompatibility complex genes, karyotype, autoimmunity, therapies, and the presence of HN exists. Of the 72 patients with TS examined, 13 had HN, a prevalence of 18.05%, which was significantly higher than in our control group (1%; P=.000001). On the contrary, only 2 patients with TS (2.77%, P=not significant) had vitiligo. By comparing the distribution of HLA class I alleles between patients with TS who did (13 of 72) and did not (59 of 72) have HN, we observed a significantly higher frequency of HLA-Cw6 in patients with TS and HN than in those without HN (26.92% vs 6.78%, respectively; P=.0067; odds ratio=5.06). The study of HLA class II genomic polymorphisms showed that the DRB1(*)0701 and DQB1*02 alleles for patients with TS and HN were overrepresented when compared with those without HN (34.61% vs 11.86%, respectively, P=.0078, odds ratio=3.93; and 34.61% vs 19.49%, respectively, P=.1386, odds ratio=2.19). In conclusion, this study is the first to demonstrate an increased prevalence of HN for patients with TS. Furthermore, the data suggest that a HN putative susceptibility gene in TS is located close to the HLA-C locus. PMID- 15337977 TI - Medallion-like dermal dendrocyte hamartoma: a new clinically and histopathologically distinct lesion. AB - Dermal dendrocyte hamartomas are extremely rare; only two examples have been described with clinical features different from our cases and with incomplete immunohistochemical characterization. We report three female patients presenting a medallion-shaped, well-defined, slightly atrophic and asymptomatic congenital lesion. All 3 patients showed a fusiform-cell proliferation. Immunohistochemistry was positive for CD34, factor XIIIa, and fascin. Electron microscopy showed typical features of dermal dendrocytes. We believe that the lesions described represent a new, clinically and histopathologically distinct lesion originating in dermal dendrocytes. We propose to name it medallion-like dermal dendrocyte hamartoma. PMID- 15337978 TI - Oral lichen planus, hepatitis C virus, and HIV: no association in a cohort study from an area of high hepatitis C virus endemicity. AB - OBJECTIVES: We sought to assess the age-specific prevalence of oral lichen planus (OLP) in Mediterranean patients with chronic hepatitis C virus (HCV) and to evaluate the features of OLP in relation to sex, smoking, HCV genotype, HIV-1 coinfection, and antiviral treatments. METHODS: In all, 178 anti-HCV-positive adults (60 women and 118 men; age range 20-66 years) recruited from two cohorts (104 HIV-negative patients and 74 patients with HIV coinfection) underwent oral examination. RESULTS: Overall prevalence of OLP was 2.8% (5 of 178) (male/female ratio 0.2; odds ratio=0.119; 95% confidence interval=0.013-1.106), only among HIV negative participants, always in the reticular pattern, and in the adult age. HCV genotype, tobacco smoking, and treatment (interferon alfa/ribavirin) were not significantly associated to OLP. CONCLUSIONS: Among HCV-infected patients, we found a low prevalence of OLP. The absence of OLP in HIV-coinfected patients suggests immunosuppression secondary to defective CD4 functions. PMID- 15337979 TI - In vivo confocal scanning laser microscopy of pigmented Spitz nevi: comparison of in vivo confocal images with dermoscopy and routine histopathology. AB - BACKGROUND: Spitz nevus is a benign melanocytic lesion sometimes mistakenly diagnosed clinically as melanoma. OBJECTIVE: Our aim was to evaluate in vivo reflectance-mode confocal scanning laser microscopy (CSLM) aspects of globular Spitz nevi and to correlate them with those of surface microscopy and histopathology. METHODS: A total of 6 Spitz nevi, with globular aspects on epiluminescence observation, were imaged with CSLM and subsequently excised for histopathologic examination. RESULTS: A close correlation among CSLM, epiluminescence, and histopathologic aspects was observed. Individual cells, observed in high-resolution confocal images, were similar in shape and dimension to the histopathologic ones. Lesion architecture was described on reconstructed CSLM images. Melanocytic nests corresponded to globular cellular aggregates at confocal microscopy and to globules at epiluminescence observation. Melanophages were clearly identified in the papillary dermis both by confocal microscopy and histopathology. CONCLUSION: In vivo CSLM enabled the identification of characteristic cytologic and architectural aspects of Spitz nevi, correlated with histopathology and epiluminescence microscopy observation. PMID- 15337980 TI - A novel connexin 26 gene mutation associated with features of the keratitis ichthyosis-deafness syndrome and the follicular occlusion triad. AB - We report the case of a congenitally deaf white male with mild palmoplantar keratoderma, ichthyosiform scaling, follicular hyperkeratosis, and mild keratitis, features consistent with keratitis-ichthyosis-deafness syndrome. His major problem was severe, disfiguring, inflammatory dissecting folliculitis of the scalp, hidradenitis suppurativa, and cystic acne, features comprising the follicular occlusion triad. This unusual phenotype is associated with a novel heterozygous point mutation (C119T) in the gap junction beta2 gene that substitutes a valine for alanine at codon 40 (A40V) in the connexin 26 protein. Through Xenopus oocyte expression studies, this mutant protein was shown to significantly disrupt the function of the specialized gap junctions connecting the cytoplasm of adjacent cells critical for tissue homeostasis. Mutations within the connexin 26 protein are associated with syndromes involving both sensorineural deafness and hyperkeratotic skin disorders. This is the first report of an association between a connexin 26 protein mutation, follicular hyperkeratosis of keratitis-ichthyosis-deafness syndrome, and severe follicular occlusion triad. PMID- 15337981 TI - Characteristics of genetic epidemiology and genetic models for vitiligo. AB - BACKGROUND: Vitiligo occurs with a frequency of 0.1% to 2% in various populations and is classified into several subtypes by its clinical presentation. Although genetic factors are thought to be involved in the cause of vitiligo, the genetic models for different phenotypes of vitiligo are unknown. OBJECTIVE: Our purpose was to explore potential genetic models for different phenotypes of vitiligo and analyze genetic epidemiologic characteristics of vitiligo in a Chinese population. METHODS: Information from 2247 patients and members in their families was collected using a uniform questionnaire. Patients' clinical characteristics and their family history were analyzed using software. A complex segregation analysis was conducted to propose potential genetic models for vitiligo. RESULTS: Different subtypes of vitiligo had different ages of disease onset. In relatives of patients with vitiligo, the risk of developing vitiligo increased with increasing relatedness to the patients with vitiligo. A polygenic additive model was the best model for focal vitiligo, vitiligo vulgaris, acrofacial vitiligo, and segmental vitiligo with approximately 50% heritability in each. For universal vitiligo, the best model was an environmental model. CONCLUSION: This study indicated that different phenotypes of vitiligo had different pathogeneses and genetic backgrounds. Onset of vitiligo is possibly affected by both genetic backgrounds and common environmental factors. PMID- 15337982 TI - Klippel-Trenaunay syndrome: the importance of "geographic stains" in identifying lymphatic disease and risk of complications. AB - BACKGROUND: Klippel-Trenaunay syndrome (KTS) is a rare congenital anomaly classically defined as the triad of vascular stain, soft tissue and/or bony hypertrophy, and venous varicosities. OBJECTIVE: To determine whether the morphologic characteristics of the associated vascular stains in KTS are predictive of the presence of lymphatic involvement and/or complications. SETTING: Outpatient dermatology practice, tertiary care medical center. METHODS: We retrospectively reviewed all cases of KTS identified between January 1989 and September 2001 at the University of California San Francisco (UCSF) Department of Dermatology. Forty patients were identified. We further classified them by type of cutaneous vascular stain, either "geographic" or "blotchy/segmental." Patients were further classified as having definite, probable, possible, or no evidence of lymphatic disease. We also reviewed the charts for other possibly associated manifestations and complications of KTS. RESULTS: Of those with sharply demarcated geographic stains (n=22), 21 had definite or probable evidence of lymphatic disease. Of those with blotchy port-wine stains (n=17), 16 had possible or no evidence of lymphatic disease (P <.001). Determination of the type of stain had 95% sensitivity and 94% specificity in differentiating the definite or probable presence of definite or probable lymphatic disease from possible or no evidence of lymphatic disease. Complications occurred in 19 (86%) of 22 patients with a geographic stain vs 7 of 17 (41%) with a blotchy/segmental stain (P <.003). CONCLUSION: This study demonstrates that the presence of a geographic vascular stain is a predictor of the risk of both associated lymphatic malformation and complications in patients with KTS. Since these stains are present at birth, this clinical observation can help in identifying individuals with KTS at greatest risk for complications and in need of closer observation. PMID- 15337983 TI - Evaluation of staging chest radiographs and serum lactate dehydrogenase for localized melanoma. AB - BACKGROUND: Radiographic and laboratory evaluations are often routinely used in the initial work-up for melanoma. PURPOSE: To examine the yield of a chest radiograph and serum lactate dehydrogenase (LDH), in the work-up for newly diagnosed localized melanoma. METHODS: Patients with a new diagnosis of localized invasive melanoma were entered into a prospective database. The status of the chest radiograph, LDH, and sentinel lymph node (SLN) was assessed. RESULTS: Two hundred-twenty-four patients were entered into the study and 210 had chest radiograph data for analysis. The true positive chest radiograph rate, defined as the percent of chest radiographs interpreted as "positive or equivocal possibly melanoma related" with subsequent confirmed melanoma metastases, was 0%. The false positive chest radiograph rate, defined as the percent of chest radiographs interpreted as "positive or equivocal possibly melanoma related" with melanoma metastases excluded based on previous or subsequent studies or other known medical conditions, was 7%. Ninety-six patients (melanoma> or =1 mm) had LDH results for analysis. Elevations in LDH were found in 15% and did not lead to detection of occult disease in any patients. Seventy-seven patients underwent SLN biopsy. A positive SLN did not correlate with abnormal chest radiograph or LDH. CONCLUSION: Low yield, high rate of false-positive tests and lack of significant impact of early detection of metastases on survival argue that chest radiographs and serum lactate dehydrogenase should probably not be accepted into routine clinical practice in patients with clinically localized melanoma in the absence of data supporting their use. PMID- 15337984 TI - Pimecrolimus 1% cream for cutaneous lupus erythematosus. AB - Topical treatment of cutaneous lupus erythematosus usually includes potent glucocorticosteroids. However, prolonged use causes adverse side effects including skin atrophy as the foremost concern. In contrast to glucocorticosteroids, the anti-inflammatory and immunosuppressive macrolactam pimecrolimus has no atrophogenic potential. Affected areas of 11 patients with different forms of lupus erythematosus were treated with pimecrolimus 1% cream under semiocclusive conditions twice daily for 3 weeks. Skin involvement before and after therapy was assessed by means of a clinical score. In all patients, significant regression of skin lesions was observed after therapy (P <.001). This was an open and uncontrolled study on a limited number of cases. We suggest that pimecrolimus 1% cream could be an efficacious and safe treatment option for cutaneous lupus erythematosus. PMID- 15337985 TI - Multiple inverted follicular keratoses as a presenting sign of Cowden's syndrome: case report with human papillomavirus studies. AB - Inverted follicular keratosis is characterized by a squamous epithelial expansion of the infundibular portion of the hair follicle in an exophytic and endophytic pattern. The lesion is often associated with squamous eddies similar to an irritated keratosis and may have a superficial papillomatous architecture. The lesion most often arises as a solitary nodule on the face of middle age to elderly individuals. Inverted follicular keratosis has been described as a distinct entity by some while others believe that it is related to trichilemmomas and/or verruca vulgares. The cutaneous pathology of Cowden's syndrome is characterized by multiple trichilemmomas. We present a woman who fulfills the clinical criteria for Cowden's syndrome and who initially presented with multiple inverted follicular keratoses. Also, in situ hybridization studies performed on the patient's keratoses do not reveal evidence of human papillomavirus infection. PMID- 15337986 TI - An update on the diagnosis and treatment of leprosy. PMID- 15337987 TI - Collagen vascular diseases. PMID- 15337988 TI - Alopecia areata investigational assessment guidelines--Part II. National Alopecia Areata Foundation. PMID- 15337989 TI - Sarcoidosis. Part I: pulmonary manifestations. PMID- 15337990 TI - Threats of biological and chemical warfare on civilian populations. PMID- 15337991 TI - Interstate dermatopathology interpretations--50 separate licenses? AB - The increasing bureaucracy of health care has had a serious negative impact on physicians and patients for the last fifteen years. We report an example of this trend involving the interstate interpretation of dermatopathology specimens, which resulted in a significant interruption of diagnostic services to a large number of dermatologists. Temporary and long term solutions are discussed with the hope that dialogue and political consensus-building can bring resolution to the problem. PMID- 15337992 TI - Surgical pearl: Cryoblast, a modified cryosurgical technique for thick lesions. PMID- 15337993 TI - Conjugal transfer vaccinia. AB - Two cases of conjugal contact transfer vaccinia are described. Each patient had intimate contact after their respective partners, active-duty military personnel, received the smallpox vaccination. PMID- 15337994 TI - Treatment of glossodynia with olanzapine. AB - A 73-year-old white male with a 6-month history of glossodynia, unresponsive to clotrimazole troches, cevimeline, triamcinolone dental paste, paroxetine, and lorazepam presented to the dermatology clinic for consultation. Work-up revealed no oral abnormalities and no underlying systemic disorder. He denied symptoms consistent with a psychiatric disorder. A detailed free amnestic assessment by a board certified Geriatric Psychiatrist (John S. Kennedy, MD) found that the patient was oppressed by the pain. He did not meet the criteria for major depression nor did he have any anxiety disorder or delusions. Because of the presence of dysphoria and anticipatory anxiety secondary to glossodynia, the patient was started on olanzapine. Improvement of pain symptoms were noted within 3 days with full resolution of symptoms at 1- and 3-month follow-ups. Dysphoria and anticipatory anxiety remitted fully upon pain relief. PMID- 15337995 TI - Invasive superficial spreading melanomas arising from clinically normal skin. AB - We provide documentation of the onset of 3 invasive superficial spreading melanomas arising from clinically normal skin and correlation with histologic lack of nevus association. Clinical, dermoscopic, and pathologic features of these "de novo" melanomas, which became invasive while small in diameter and had few clinical criteria for diagnosis, are reported. PMID- 15337996 TI - Howard Fox, MD, founding father, American Academy of Dermatology: a historical tribute on the 50th anniversary of his death. PMID- 15337997 TI - Sweet's syndrome following abacavir therapy. PMID- 15337998 TI - Treatment of Hailey-Hailey disease with topical calcitriol. PMID- 15337999 TI - Cutaneous presentation of ectopic small bowel mucosa. PMID- 15338000 TI - Angioedema associated with imiquimod. PMID- 15338001 TI - Molluscum folliculitis after leg shaving. PMID- 15338002 TI - Comment on the psoriasis periodic synopsis. PMID- 15338003 TI - Thalidomide is ineffective in the treatment of lichen planopilaris. PMID- 15338004 TI - Methotrexate and reactivation tuberculosis. PMID- 15338005 TI - Optimizing bexarotene therapy for cutaneous T-cell lymphoma. PMID- 15338006 TI - Should multiple lymph node biopsies be taken? PMID- 15338008 TI - Heterozygous deletion of the linked genes ZIC1 and ZIC4 is involved in Dandy Walker malformation. AB - Dandy-Walker malformation (DWM; OMIM #220200) is a common but poorly understood congenital cerebellar malformation in humans. Through physical mapping of 3q2 interstitial deletions in several individuals with DWM, we defined the first critical region associated with DWM, encompassing two adjacent Zinc finger in cerebellum genes, ZIC1 and ZIC4. Mice with a heterozygous deletion of these two linked genes have a phenotype that closely resembles DWM, providing a mouse model for this malformation. PMID- 15338009 TI - The contribution of spike threshold to the dichotomy of cortical simple and complex cells. AB - The existence of two classes of cells, simple and complex, discovered by Hubel and Wiesel in 1962, is one of the fundamental features of cat primary visual cortex. A quantitative measure used to distinguish simple and complex cells is the ratio between modulated and unmodulated components of spike responses to drifting gratings, an index that forms a bimodal distribution. We have found that the modulation ratio, when derived from the subthreshold membrane potential instead of from spike rate, is unimodally distributed, but highly skewed. The distribution of the modulation ratio as derived from spike rate can, in turn, be predicted quantitatively by the nonlinear properties of spike threshold applied to the skewed distribution of the subthreshold modulation ratio. Threshold also increases the spatial segregation of ON and OFF regions of the receptive field, a defining attribute of simple cells. The distinction between simple and complex cells is therefore enhanced by threshold, much like the selectivity for stimulus features such as orientation and direction. In this case, however, a continuous distribution in the spatial organization of synaptic inputs is transformed into two distinct classes of cells. PMID- 15338010 TI - E1A-induced apoptosis does not prevent replication of adenoviruses with deletion of E1b in majority of infected cancer cells. AB - Apoptotic pathways are initiated as a cellular defense mechanism to eliminate adenovirus-infected cells. We have investigated how E1A-induced apoptosis interferes with viral replication in cancer cells. We found that E1B19K alone can efficiently suppress E1A-induced apoptosis in cancer cells. Viruses deleted for both E1B19K and E1B55K resulted in cellular DNA degradation. However, less than 20% of human lung cancer cells infected with a virus deleted for both E1B19K and E1B55 K had evidence of chromatin condensation and multiple-micronuclei formation (apoptotic hallmarks); these cells could not produce infectious viral particles. The majority of cancer cells infected with viruses deleted for the entire E1b gene did not undergo extended apoptosis and produced abundant viral progeny. Thus, only a fraction of cancer cells underwent apoptosis and did not allow E1b deleted viruses to replicate, while the majority of cancer cells were resistant to E1A-induced apoptosis and could support virus-selective replication. The results of this study imply that, in addition to inhibiting E1A-induced apoptosis, E1B proteins may contribute other important roles in the viral life cycle. Our results also suggest that combining virus-induced apoptosis and selective viral replication into one vector will be a novel approach to destroy cancer cells. PMID- 15338011 TI - Synergistic antitumor effect of antiangiogenic factor genes on colon 26 produced by low-voltage electroporation. AB - Antiangiogenic factors are potent endothelial cell growth inhibitors that have been shown to inhibit angiogenesis in vitro and tumor growth in mice. We have demonstrated the synergistic antitumor effect of antiangiogenic genes (mouse angiostatin: pBLAST-mAngio; and mouse endostatin: p-BLAST42-mEndo XV) delivered to tumors by low-voltage electroporation in mouse colon 26 models. A synergistic antitumor effect was strongly suggested by in vivo tumor growth kinetics, as well as in survival studies with the mice. RT-PCR confirmed that the fragments of each gene were transferred by low-voltage electroporation in the tumor. Decreased microvessel density measurements in tumors also confirmed the efficacy of the synergistic antitumor effect of both genes. Significant growth inhibition was observed in mice treated with a 1:1 proportion of angiostatin and endostatin genes, and the order of the both genes transferred (first the endostatin gene, followed 1 week later by the angiostatin gene) had a profound inhibitory effect on tumor growth. These data suggest that in vivo delivery of antiangiogenic genes with low-voltage electroporation could be a possible therapeutic strategy for established solid tumors when both genes were applied in combination. PMID- 15338012 TI - Genetic factors in hypertension. Angiotensin-converting enzyme polymorphism. AB - AIM: The key enzyme of the renin-angiotensin-aldosterone system angiotensin converting enzyme (ACE), shows the genetic polymorphism responsible for the varying activity of this enzyme. In a study of randomly chosen families we analyzed the relationship between insertion/deletion (I/D) polymorphism of the ACE and ambulatory blood pressure (ABP), left ventricular mass index (LVMI) and carotid intima-media thickness (IMT). METHODS: The study population consisted of 127 parents and 167 offspring. All subjects underwent 24 hr ABP monitoring using a SpaceLabs 90207 device. 2D and M-mode echocardiograms were also obtained. The carotid intima-media thickness (IMT) was assessed by ultrasound. The I/D polymorphism of the ACE gene was measured with the use of PCR method. For statistical analysis, co-variables and correlations between relatives were taken into account. RESULTS: The frequency of genotypes was: 27.2% for DD homozygotes, 49.7% for DI heterozygotes and 23.1% for II homozygotes, being in Hardy-Weinberg equilibrium (P=0.97). There was no relationship between the ACE gene polymorphism and ABP, LVMI or carotid IMT in parents nor their offspring. The transmission disequilibrium tests for quantitative traits showed only a slight tendency towards the influence of the polymorphism on LVMI (P=0.06). CONCLUSIONS: In the study group of nuclear families, I/D polymorphism of the ACE gene did not influence blood pressure, left ventricular mass or carotid intima-media thickness. PMID- 15338013 TI - Myocardial infarction and patent coronary arteries--an uneventful association? AB - BACKGROUND: Although many studies addressed the issue of the occurrence of acute myocardial infarction (MI) in patients with angiographically patent coronary arteries, controversies exist concerning the pathophysiology and clinical outcome in this syndrome. AIM: To evaluate the clinical course of patients with MI and patent coronary arteries as well as to assess the post-infarction short-term prognosis. METHODS: A retrospective analysis of patients with acute MI admitted to our institution over a period of 8 years (1995-2002) was performed. The study group consisted of 62 patients with MI and angiographically patent coronary arteries, examined within the first 30 days after the infarction, and the age and gender-matched control group of 62 patients with acute MI and significant coronary artery stenoses. RESULTS: Cardiovascular risk factors found in the study group versus the control group were: smoking 47 (66.1%) vs 54 (87.1%) patients (NS); dyslipidemia 18 (29.0%) vs 44 (74.2%) patients (p<0.005), and family history of coronary artery disease 16 (25.8%) vs 37 (61.3%) patients (p<0.05). The post-infarction complications were: ischaemic recurrences in 25 (40.3%) vs 46 (74.2%) patients (p<0.05); heart failure in 11 (17.7%) vs 26 (41.9%) patients (p<0.05); mechanical events in 12 (19.4%) vs 16 (25.8%) patients (NS); arrhythmias in 18 (29.0%) vs 12 (19.4%) patients (NS); peripheral thromboembolic complications in 5 (8.1%) vs 2 (3.2%) patients (NS), respectively. CONCLUSIONS: In the population with MI and patent coronary arteries, smoking was the only well represented risk factor. Although there was a significantly lower incidence of ischaemic events and heart failure after MI in patients with patent rather than stenotic coronary arteries, the mechanical, arrhythmic and thromboembolic complications occurred with the same frequency. PMID- 15338014 TI - Proinflammatory cytokine levels in patients with diastolic heart failure. AB - BACKGROUND: The role of cytokines in the pathogenesis of systolic heart failure (HF) has been well established whereas in diastolic HF it remains uncertain. AIM: To define levels of Tumor Necrosis Factor-a (TNFa) and Interleukin-6 (IL-6) in patients with diastolic HF and to reveal their association with functional class and types of left ventricular (LV) diastolic dysfunction. METHODS: We examined 26 patients with diastolic HF. The control group consisted of 10 healthy persons. Commonly used echocardiographic parameters of systolic and diastolic function of the LV, thickness of the interventricular septum (IVS), posterior LV wall thickness (PWLV), end-diastolic size of the LV (EDSLV) and left atrial (LA) volume were assessed. Serum levels of TNFa and IL-6 were measured with highly sensitive enzyme-linked immunosorbent assay. RESULTS: The TNFa and IL-6 levels were significantly higher in the group with diastolic HF than in the control group. TNFa and IL-6 levels in groups with impaired LV relaxation and restriction/pseudonormalisation were significantly higher than in the control group. TNFa level was significantly higher in the restriction/pseudonormalisation group than in the group with impaired relaxation, whereas the IL-6 level was similar. The TNFa level was significantly higher in the group with NYHA class III IV in comparison to the group with NYHA class II. The IL-6 level in these groups was similar. Of the echocardiographic parameters, only LA volume significantly correlated with the TNF+/- level. No relationship between the IL-6 level and echocardiographic parameters was found. CONCLUSIONS: In diastolic HF, serum levels of TNFa and IL-6 are elevated. The magnitude of TNF+/- elevation is associated with the severity of HF, assessed by NYHA classification, LV diastolic dysfunction level or LA volume. PMID- 15338015 TI - Heart rate variability in workers chronically exposed to lead. AB - BACKGROUND: Lead is a strong neurotoxin. The effects of lead on the activity of the autonomic nervous system, assessed by the use of heart rate variability (HRV) analysis, have not yet been established. AIM: To assess the effects of occupational chronic lead exposure on the autonomic nervous system activity. METHODS: The study group consisted of 22 copper foundry workers (mean age 41.8+/ 8.7 years) who had elevated parameters of lead overload and were admitted to the hospital for chelate therapy. The control group consisted of 13 age-matched healthy males. Lead concentration was measured with the use of atomic absorption spectrophotometry, and concentration of free protoporphyrins in erythrocytes (FEP) using a fluorometric method. Each patient underwent 24-hour ambulatory ECG monitoring, and standard short-term as well as long-term HRV parameters were obtained. RESULTS: There were no significant differences between patients and controls in HRV parameters. In the control group, HRV parameters correlated with age. In patients, a significant negative correlation between lead concentration and some short-term HRV parameters calculated during the night was found: SDNN (r=-0.48, p<0.05), TP (r=-0.48, p<0.01) and LF (r=-0.48, p<0.01). In patients, a negative correlation between lead concentration and HFnight/HFday index was found (r=-0.47 p<0.01), whereas in controls this correlation was positive (r=0.66 p<0.05). CONCLUSIONS: Overall HRV indices are similar in subjects exposed to lead and in healthy controls. A decrease in the physiological elevation of HF values during the night, together with an increase in lead blood concentration and lack of relationship between age and HRV parameters in workers chronically exposed to lead may suggest disturbances of the autonomic system. In subjects not exposed to lead a decrease in heart rate with an increase in FEP concentration was observed. PMID- 15338016 TI - Transcatheter closure of perimembranous ventricular septal defect using an Amplatzer occluder--early results. AB - BACKGROUND: Transcatheter closure of perimembranous ventricular septal defect with the use of an Amplatzer occluder was introduced in 1999, however, clinical experience with this method remains limited. AIM: To assess short-term safety and efficacy of this technique in children. METHODS: Since 2001, nine children underwent transcatheter closure of perimembranous ventricular septal defect in the cardiac centre in Bratislava (Slovak Republic) or in our centre. Patients were selected for the procedure based on the results of transesophageal echocardiography (TEE). Transcatheter closure using the Amplatzer occluder was guided by both TEE and angiography. The follow-up duration ranged from 3 to 22 months, mean 11.5 months. Control angiography was performed in order to detect any leakage through the aortic valve. The end-diastolic left ventricular diameter before and after the procedure as well as the function of the aortic, tricuspid and mitral valves were also assessed. RESULTS: The procedure was effective in all patients. No damage to atrio-ventricular nor aortic valves was observed. Tricuspid regurgitation, present prior to the procedure, markedly decreased following the closure of the defect. The left ventricular end-diastolic diameter significantly (p=0.001) decreased after the procedure. CONCLUSIONS: (1) Perimembranous ventricular septal defect can be safely and effectively closed using a transcatheter Amplatzer occluder. (2) Appropriate selection for the procedure as well as TEE and angiographic guidance during the procedure enable the avoidance of complications. (3) Tricuspid regurgitation is not an absolute contraindication to perform this procedure. PMID- 15338017 TI - [Cost-effectiveness of coronary angioplasty procedures in Poland]. AB - BACKGROUND: Restenosis following percutaneous coronary interventions (PCI) increases re-hospitalisation rate and may lead to new myocardial infarction (MI) or death. Besides medical aspects, it may also reduce cost-effectiveness of the procedure. AIM: To analyse the medical and economical outcome of patients treated with PCI during a one year period. METHODS: Medical outcome, cost of PCI and total cost of treatment during one year after PCI were assessed in 188 consecutive patients who underwent PCI during the first three months of 2002. Patients with acute MI treated with PCI were not included in the analysis. RESULTS: The rate of major adverse cardiac events (MACE) which included death, new MI or repeated revascularisation, was 1.6% during hospital stay and 14.4% during one-year follow-up. Re-hospitalisation rate was 28.2%. The mean number of outpatient visits during one year was 9.8. The costs of initial hospitalisation and procedures performed during this hospital stay were 7,839 Polish zlotys (PLN) per patient whereas the costs during one-year follow-up were 3,490 PLN (re hospitalisations and repeated procedures 3,091 PLN, outpatient visits 238 PLN, and pharmacotherapy costs 161 PLN). In the group of patients with MACE, the costs of treatment during one-year follow-up were 13,398 PLN whereas in patients without complications 1,349 PLN per patient. CONCLUSIONS: Patients who develop complications after PCI generate costs exceeding ten times that of patients with a favourable outcome. Thus, from the economical and medical point of view, there is a need to identify high-risk patients before the decision is made as to which type of treatment is used. Because the health service in Poland is under-funded, patients at risk should be treated with the most effective methods (antiproliferative stents or surgical revascularisation) as an initial treatment, which may decrease total costs during a long-term period. PMID- 15338018 TI - [Recurrent thrombosis of a mitral mechanical heart valve prosthesis during puerperium--a case report]. AB - A case of a 27-year-old female with prosthetic mitral valve is presented. Prolonged anticoagulation therapy was continued during pregnancy without complications. During puerperium, the dose of subcutaneous low molecular weight heparin was reduced due to subcutaneous blood effusions. Subsequently, the patient developed acute left ventricular heart failure due to prosthetic valve thrombosis. She underwent urgent surgery with new prosthetic valve implantation. Two weeks later she suffered another episode of acute mitral prosthetic valve thrombosis which was effectively treated with intravenous heparin. Difficulties concerning prolonged anticoagulation during pregnancy and puerperium in patients with prosthetic valves are discussed. PMID- 15338019 TI - [Plethysmographic assessment of pulse wave may be helpful in the detection of prosthetic valve dysfunction--a case report]. AB - A case of a 63-year-old female with prosthetic mitral and aortic valves is described. The long-term post-operative period was complicated by infective endocarditis, persistent atrial fibrillation requiring pacemaker implantation and total a-v node ablation as well as ischaemic stroke which occurred one year before present hospitalisation. This time the patient was admitted to the hospital due to progressive heart failure. Transesophageal echocardiography showed a cyclic intermittent opening of both prosthetic valves full opening was present during every second cardiac cycle. The same phenomenon was documented using plethysmographic recording of a pulse wave from a finger. The patient underwent prosthetic valve replacement. Intraoperatively, a fibrous tissue ingrowth was detected. PMID- 15338020 TI - [Congenitally corrected transposition of the great arteries in a 72 year old man- a case report]. AB - Congenitally corrected transposition of the great arteries (CCTGA) is a rare form of congenital heart disease characterised by atrioventricular as well as ventriculoarterial discordance. The life expectancy of individuals with CCTGA is limited by the onset of the systemic ventricular failure. There have been only a few patients with CCTGA and age >50 years reported in literature. We describe a 72-year-old man with CCTGA who was admitted to the hospital because of severe congestive heart failure. Our patient s survival is one of the longest ever described. PMID- 15338021 TI - [Implantation of an automatic cardioverter-defibrillator in small children--two case reports]. AB - Implantation of an automatic cardioverter-defibrillator (ICD) in children may be challenging due to the increased risk of periprocedural and long-term complications. ICD was implanted in two boys with hypertrophic cardiomyopathy, aged 6 and 9 years, with of a body weight of 20 and 25 kg, respectively. In one patient an ICD was implanted due to a history of ventricular fibrillation whereas the second patient underwent prophylactic ICD implantation due to a family history of sudden cardiac death. No short- or mid-term complications were recorded. Difficulties and risks of ICD implantation in children are discussed. PMID- 15338022 TI - [Cardiac amyloidosis--an update]. PMID- 15338024 TI - [Abnormal mass localised on an anterior mitral leaflet chordae tendinae--a potential cause of stroke in a 39 year old female]. PMID- 15338025 TI - [Electrocardiogram of the month]. PMID- 15338026 TI - [Angiogram of the month: angina pectoris]. PMID- 15338027 TI - [Prevention of cardiovascular diseases. Is red wine more effective than other alcoholic beverages?]. PMID- 15338030 TI - Intriguing olfactory proteins from the yellow fever mosquito, Aedes aegypti. AB - Four antennae-specific proteins (AaegOBP1, AaegOBP2, AaegOBP3, and AaegASP1) were isolated from the yellow fever mosquito, Aedes aegypti and their full-length cDNAs were cloned. RT-PCR indicated that they are expressed in female and, to a lesser extent, in male antennae, but not in control tissues (legs). AaegOBP1 and AaegOBP3 showed significant similarity to previously identified mosquito odorant binding proteins (OBPs) in cysteine spacing pattern and sequence. Two of the isolated proteins have a total of eight cysteine residues. The similarity of the spacing pattern of the cysteine residues and amino acid sequence to those of previously identified olfactory proteins suggests that one of the cysteine-rich proteins (AaegOBP2) is an OBP. The other (AaegASP1) did not belong to any group of known OBPs. Structural analyses indicate that six of the cysteine residues in AaegOBP2 are linked in a similar pattern to the previously known cysteine pairing in OBPs, i.e., Cys-24-Cys-55, Cys-51-Cys-104, Cys-95-Cys-113. The additional disulfide bridge, Cys-38-Cys-125, knits the extended C-terminal segment of the protein to a predicted alpha2-helix. As indicated by circular dichroism (CD) spectra, the extra rigidity seems to prevent the predicted formation of a C terminal alpha-helix at low pH. PMID- 15338031 TI - Month of birth and life expectancy: role of gender and age in a comparative approach. AB - The effects of month of birth (MOB) on life expectancy of a German subpopulation was investigated. Data from people who died in North Rhine Westphalia in the years 1984 ( n=188,515) and 1999 ( n=188,850) were analyzed. For comparative purposes, all deaths that occurred at an age of <50 years were excluded (1984: 8.4%; 1999: 6.2%). In general, individuals born in May through July had the lowest age at death (1984: 75.27+/-0.09 years; 1999: 77.58+/-0.09 years), while those born between October and December had the highest (1984: 75.98+/-0.08 years; 1999: 78.35+/-0.09 years), supporting earlier findings. The observed amplitudes (differences between highest and lowest values) were more pronounced in men than in women. When comparing these data of MOB effects on life expectancy with earlier findings in Australia, Austria, Denmark, Ukraine, and the USA, it is evident that a negative correlation exists between the average age at death and the MOB amplitudes. Separate analyses by gender, possible for the data from Germany, the Ukraine, and the USA, revealed a significant negative correlation for men, but not for women. A new hypothesis is therefore presented describing an influence of life quality, as reflected by average life expectancy, on the extent of MOB effects; for example, seasonally variable sensitivities during pregnancy/early childhood. PMID- 15338032 TI - Transgenic insect-resistant corn affects the fourth trophic level: effects of Bacillus thuringiensis-corn on the facultative hyperparasitoid Tetrastichus howardi. AB - As hyperparasitoids may have a considerable influence on the control of herbivorous arthropods, analyzing the host-mediated impact of Bacillus thuringiensis-plants ("Bt-plants") on hyperparasitoids is of interest. Laboratory scale experiments were conducted in order to assess the potential effect of Bt corn leaf material on the facultative hyperparasitoid Tetrastichus howardi (Olliff) (Hymenoptera: Eulophidae), mediated through the herbivore Chilo partellus Swinhoe (Lepidoptera: Crambidae) and its primary parasitoid Cotesia flavipes (Cameron) (Hymenoptera: Braconidae). In the Bt-group, significantly less C. flavipes larvae per host spun cocoons and pupated than in the control, and the mean fresh weight of a single C. flavipes cocoon was significantly reduced compared to the control. All C. flavipes cocoons of one host formed cocoon clusters. T. howardi females of the Bt-group parasitized significantly less cocoon clusters than in the control. Moreover, significantly fewer C. flavipes cocoons per cocoon cluster were successfully parasitized as compared to the control. As a consequence, T. howardi females of the control had more offspring than in the Bt-group. Adult female T. howardi offspring of the Bt-group weighed significantly less than in the control, but there was no significant weight difference between males of both groups. Our results suggest that transgenic insect-resistant plants could affect hyperparasitoids indirectly. However, it remains to be determined whether facultative hyperparasitoids prefer to develop as primary or secondary parasitoids under field conditions. PMID- 15338033 TI - [The minimally invasive stabilization of the dorsal pelvic ring with the transiliacal internal fixator (TIFI)--surgical technique and first clinical findings]. AB - The Trans Iliacal Internal Fixator (TIFI) is a minimally invasive technique for the stabilization of sacro-iliac joint ruptures and fractures lateral to the sacral ala or through the sacral foramen. In this study, 7.0 mm pedicle screws of the Universal-Spine-System (USS, Synthes) were inserted 1-2 cm on the cranial side of the posterior superior iliac spine and parallel to the superior gluteal line. The connecting bar was inserted subfascially and fixed with the locking head pedicle screws to form an fixed-angle construction. In a prospective study 31 patients with vertical shear injuries of the pelvis were treated with the TIFI. There were two wound infections and one loosening of a pedicle screw. None of the screws were incorrectly positioned and no neuro-vascular lesions were caused by the implant. 2 years postoperatively we found 50% good and excellent results for type C pelvic ring injuries. Early findings show that the TIFI is well suited to stabilization of sacro-iliac joint ruptures and fractures of the lateral sacrum. Closed reduction and minimally invasive insertion technique are possible. The implant leads to sufficient biomechanical stability but there is a very low intraoperative risk of neuro-vascular lesion. PMID- 15338034 TI - High variability of peptidylarginine deiminase 4 (PADI4) in a healthy white population: characterization of six new variants of PADI4 exons 2-4 by a novel haplotype-specific sequencing-based approach. AB - Seven single nucleotide polymorphisms (SNPs) of the peptidylarginine deiminase 4 (PADI4) gene have recently been reported to be strongly associated with rheumatoid arthritis in Japanese individuals. These SNPs are located in or close to exons 2-4 of PADI4 and are organized in at least four different haplotypes. However, a detailed sequencing-based characterization of the PADI4 gene in other populations is still lacking. We therefore analyzed exons 2-4 of the PADI4 gene in 102 healthy white Germans individuals by DNA sequencing and characterized new variants and haplotypes by a novel haplotype-specific sequencing-based approach. The haplotypes 2/3 (padi4_89*G, padi4_90*T, padi4_92*G, padi4_94*T, padi4_104*T, padi4_95*C, padi4_96*C), and haplotype 4 (padi4_89*G, padi4_90*T, padi4_92*G, padi4_94*T, padi4_104*C, padi4_95*G, padi4_96*T) conferring susceptibility to rheumatoid arthritis were detected at frequencies of 30.9% and 7.8%, respectively. In addition, three novel coding SNPs in exons 2, 3, and 4, and three SNPs in introns 2 and 3 located near the exon-intron boundaries were identified in 11 individuals (10.8%). The so-called nonsusceptibility haplotype 1 (padi4_89*A, padi4_90*C, padi4_92*C, padi4_94*C, padi4_104*C, padi4_95*G, padi4_96*T) occurred at a frequency of 58.3%. Additionally, we identified a closely related novel haplotype, haplotype 1B (2.9%), that differs from haplotype 1 only by padi4_92*G/padi4_96*C. This haplotype was not described in the Japanese population. Our results indicate that the PADI4 gene exhibits a remarkable variability and a rather complex haplotypic organization. Further studies on disease association of PADI4 should be performed by haplotype-specific sequencing based approaches to identify the exact genotype of the PADI4 fragment of interest. PMID- 15338035 TI - [Functional dynamics of the right ventricle and pulmonary circulation in obstructive sleep apnea. Therapeutic consequences]. AB - Obstructive sleep apnea (OSA) is common with an incidence of at least 500,000 patients in the German population. Typical symptoms are daytime sleepiness, headache in the morning, and snoring. Presumably obstructive sleep apnea via various mechanisms increases cardiovascular morbidity. Hypoxemia causes nocturnal hypertension in most of the patients. Nevertheless, about 20% of the patients develop daytime pulmonary hypertension and right heart dysfunction. Clinical and animal studies demonstrated right ventricular hypertrophy as a consequence of intermittent hypoxemia and pulmonary hypertension. Right ventricular hemodynamics differ essentially from left ventricular hemodynamics. Right ventricular function is substantially influenced by right ventricular afterload, which is mainly determined by pulmonary vascular resistance, and slightly influenced by preload. Application of continuous positive airway pressure (CPAP) via a nose mask normalizes nocturnal breathing disorders and reduces pre- and afterload, especially in patients with cardiomegaly. Therefore, CPAP generates positive effects on the myocardium. PMID- 15338036 TI - [Right ventricular arrhythmias]. AB - Right ventricular arrhythmias predominantly occur in young patients with rare cardiac diseases. Underlying cardiac conditions include idiopathic right ventricular outflow-tract tachycardia (RVOT-VT), arrhythmogenic right ventricular cardiomyopathy (ARVC), Brugada syndrome, and postoperative congenital heart disease (i.e. tetralogy of Fallot). According to the underlying cardiac disease, there are significant differences in the diagnostic and therapeutic management and prognosis which is mainly determined by life-threatening ventricular arrhythmia recurrences and sudden cardiac death. To provide optimal treatment for affected patients, a detailed diagnostic evaluation and risk stratification is mandatory. Tailored treatment strategies aim at the suppression or effective termination of recurrent ventricular tachyarrhythmias and prevention of sudden death by antiarrhythmic drug therapy, catheter ablation, and the implantation of cardioverter-defibrillators. This review summarizes the current knowledge on pathogenesis, diagnosis, treatment and prognosis of those conditions that are associated with arrhythmias originating from the right ventricle. PMID- 15338037 TI - [Fibrin glue for operative correction of septal deviations]. AB - The routine procedure after correction of septal deviations is the utilization of endonasal packing in order to avoid septal haematoma. However, the mechanical pressure of this packing damages the mucociliar activity of the mucosa and causes lymphoedema by blocking the lymphatic vessels. Besides it represents a foreign body within the nose causing pain and unpleasant feeling for the patient. In order to avoid these disadvantages, in 57 patients we used fibrin glue instead of nasal packing. After correction of the septal deviation the two mucoperichondrium blades where fixed together with Tissucol Duo Quick. This technique not only leads to haemostasis, but also provides fixation of the newly modeled septum. In only one patient we found a small unilateral septal haematoma, in all other cases the postoperative period showed no complications. Patients had a significant reduction of endonasal crusts and postoperative swelling. Patients comfort increased significant without the (standard) nasal packing. PMID- 15338038 TI - [Function tests for the Eustachian tube. Current knowledge]. AB - The Eustachian tube regulates the homeostasis of the middle ear. Problems with its function are predominantly found in childhood. As a consequence, otitis and hearing impairment occur. The most important muscle is the tensor tympani muscle. The complexity of the functional anatomy and physiology are reasons why no function test alone is capable of fulfilling all diagnostic needs. The predictive value of the various Eustachian tube function tests for the outcome of a tympanoplasty is not yet clear. An overview of clinically and scientifically relevant tests for the Eustachian tube function is provided. PMID- 15338040 TI - [Those said to be dead live longer...]. PMID- 15338041 TI - [In the mills of justice...suspension of a clinic director lifted--decision of the administrative tribunal Giessen, 6/21/2004]. PMID- 15338042 TI - Coordination of epigenetic events. AB - During the course of DNA damage a complex repertoire of molecular signals, chromatin determinants and specific transcription factors are set in motion for repair. In many instances, the response pathway can be characterized by profound changes in molecular remodeling and is intimately linked with DNA replication and gene transcription. Our understanding of the molecular pathways has come from scientific developments that represent many disparate disciplines, such as cancer (epi)genetics, chromatin modifications during cellular development and the emerging prominence of epigenetic events in human disease. These multidisciplinary areas reveal a functional relationship and suggest that repair and transcription must coincide in the context of chromatin. We have come to appreciate the repair process and the role of transcriptional components in a sophisticated program of epigenetic regulation, and we have learnt much since the first description of the nucleosome as a spheroid disklike unit. The coordinated and ordered response to DNA damage can specify structures that mobilize and remodel nucleosomes. Investigators will undoubtedly continue to explore the structural and functional states of DNA damage repair and continue to profile the sequence of events and scrutinize the molecular signatures that specify these changes in chromatin dynamics, genomic stability and transcriptional performance. In this special issue, authors have contributed reviews that discuss hypotheses and results regarding DNA damage repair and transcription. The topics covered range from DNA repair in a chromatin environment to the deadly double-strand break, histone modifications to ATP-dependent chromatin remodeling, gene silencing in cancer to apoptosis and regulation of chromatin dynamics by DNA methylation. The scene is set for a new view of damage detection and repair by the coordination of epigenetic states. PMID- 15338043 TI - Double-strand breaks: signaling pathways and repair mechanisms. AB - Double-strand breaks arise frequently in the course of endogenous - normal and pathological - cellular DNA metabolism or can result from exogenous agents such as ionizing radiation. It is generally accepted that these lesions represent one of the most severe types of DNA damage with respect to preservation of genomic integrity. Therefore, cells have evolved complex mechanisms that include cell cycle arrest, activation of various genes, including those associated with DNA repair, and in certain cases induction of the apoptotic pathway to respond to double-strand breaks. In this review we discuss recent progress in our understanding of cellular responses to DNA double-strand breaks. In addition to an analysis of the current paradigms of detection, signaling and repair, insights into the significance of chromatin remodeling in the double-strand break-response pathways are provided. PMID- 15338044 TI - DNA repair in a chromatin environment. AB - DNA mutations and aberrations are a problem for all forms of life. Eukaryotes specifically have developed ways of identifying and repairing various DNA mutations in a complex and refractory chromatin environment. The chromatin structure is much more than a packaging unit for DNA; it is dynamic. Cells utilize and manipulate chromatin for gene regulation, genome organization and maintenance of genome integrity. Once a DNA aberration has occurred, the various DNA repair machineries interact with chromatin proteins, such as the histone variant H2A.X, and chromatin remodeling machines of the SWI/SNF family to gain access and repair the lesion in a timely manner. Recent studies have thus begun to address the roles of chromatin proteins in DNA repair as well as to dissect the functions of DNA repair machinery in vitro on more physiological, nucleosomal templates. PMID- 15338045 TI - Chromatin modifiers in transcription and DNA repair. AB - In eukaryotes, the packaging of DNA into nucleosomes and the organization of chromatin fibres generate constraints for all nuclear processes involving DNA, including replication, repair, recombination and transcription. The three major processes that regulate chromatin structure and counterbalance its repressive effects are ATP-dependent chromatin remodeling, post-translational modification of histones and histone replacement. While many of these processes have been intensively studied with respect to their effects on transcription, there is also evidence that they affect other nuclear processes involving DNA. This review explores the functions of chromatin-remodeling factors and histone-modifying enzymes in gene regulation and summarizes recent findings which suggest a role for these chromatin modifiers in DNA repair. PMID- 15338046 TI - Chromosomal regulation by MeCP2: structural and enzymatic considerations. AB - The unique properties of eukaryotic DNA modified via methylation of cytosine residues are believed to result from the action of a conserved family of proteins, the MBD family. The prototype member of this family, MeCP2, was isolated independently in two laboratories. One group isolated MeCP2 as a methylated DNA-binding protein, the second as a sequence-specific DNA-binding protein. Multiple lines of evidence suggest that MeCP2 functions in assembly of specialized chromatin architecture. While initial findings pointed to an enzymatic mechanism involving histone modification for transcriptional repression mediated by MeCP2, emerging studies clearly provide exceptions to this model. In a recent study, highly compacted, unique chromatin structures were generated by stoichiometric binding of MeCP2 to model chromatin fibers. These findings support the likelihood that MeCP2 can utilize two independent, but not mutually exclusive, mechanisms to repress transcription: enzymatic and structural mechanisms. PMID- 15338047 TI - Gene silencing in DNA damage repair. AB - Silencing of DNA repair genes plays a critical role in the development of the cancer because these genes, functioning normally, would prevent the accumulation of mutations leading to carcinogenesis. Epigenetic gene silencing is an alternative mechanism to genetic gene aberration, inactivating those genes in cancer. DNA methylation and histone modification are the major factors for epigenetic regulation of gene expression. Here, we describe recent advances in understanding of epigenetic silencing of DNA repair genes and their epigenetic mechanisms involving DNA methylation and histone modification. PMID- 15338048 TI - Functional links between transcription, DNA repair and apoptosis. AB - DNA damage initiates damage response pathways, cell cycle arrest and apoptosis. These processes act in a concerted fashion and remain functionally linked through mechanisms not completely understood. Programmed cell death, referred to as apoptosis, is a tightly regulated phenomenon ensuring that cells that accumulate irreversible DNA damage do not replicate. Interestingly, hyperacetylation of histone proteins, which alters transcription patterns and appears linked to DNA repair, also induces apoptosis, suggesting that aspects of chromatin modification link these very distinct processes. Modulating chromatin structure in the absence of any DNA lesions also activates key DNA damage-signalling proteins, further supporting the role of higher-order chromatin structure in mediating stress responses. This review will present an overview of the epigenetic control of eukaryotic genomes by chromatin remodelling as it pertains to DNA damage and highlight the potential role of the ING PHD proteins in linking apoptosis and DNA repair to gene transcription. PMID- 15338049 TI - Focusing on phosphomonohydrolases: a timely choice. PMID- 15338050 TI - Lanthanide-binding peptides and the enzymes that Might Have Been. AB - The trivalent lanthanide ions are chemically similar to Ca(II) ions, making them useful Ca analogs for a multitude of applications. In addition, Ln(III) ions are efficient catalysts of hydrolysis due to their much stronger Lewis acidity relative to Ca(II) ions. Ln-binding peptides thus offer both the opportunity to study known Ca sites as well as to explore new biological functions with an entire family of spectroscopically rich and reactive ions. This review discusses Ln-binding peptides in three roles: (i) as models of Ca-protein structure and function, (ii) as spectroscopic tags for protein expression and characterization and (iii) as designed artificial endonucleases. The creation of hydrolytically active Ln peptides that can fold, bind, cleave and discriminate among substrates shows that the design of Ln enzymes can be accomplished, and they will serve as versatile biochemical tools to investigate protein folding, structure and nuclease function. PMID- 15338051 TI - Alternative splicing of Bcl-2-related genes: functional consequences and potential therapeutic applications. AB - Apoptosis is a morphologically distinct form of cell death. It is executed and regulated by several groups of proteins. Bcl-2 family proteins are the main regulators of the apoptotic process acting either to inhibit or promote it. More than 20 members of the family have been identified so far and most have two or more isoforms. Alternative splicing is one of the major mechanisms providing proteomic complexity and functional diversification of the Bcl-2 family proteins. Pro- and anti-apoptotic Bcl-2 family members should function in harmony for the regulation of the apoptosis machinery, and their relative levels are critical for cell fate. Any mechanism breaking down this harmony by changing the relative levels of these antagonistic proteins could contribute to many diseases, including cancer and neurodegenerative disorders. Recent studies have shown that manipulation of the alternative splicing mechanisms could provide an opportunity to restore the proper balance of these regulator proteins. This review summarises current knowledge on the alternative splicing products of Bcl-2-related genes and modulation of splicing mechanisms as a potential therapeutic approach. PMID- 15338052 TI - Resistance to beta-lactam antibiotics. AB - beta-lactams have a long history in the treatment of infectious diseases, though their use has been and continues to be confounded by the development of resistance in target organisms. beta-lactamases, particularly in Gram-negative pathogens, are a major determinant of this resistance, although alterations in the beta-lactam targets, the penicillin-binding proteins (PBPs), are also important, especially in Gram-positive pathogens. Mechanisms for the efflux and/or exclusion of these agents also contribute, though often in conjunction these other two. Approaches for overcoming these resistance mechanisms include the development of novel beta-lactamase-stable beta-lactams, beta-lactamase inhibitors to be employed with existing beta-lactams, beta-lactam compounds that bind strongly to low-affinity PBPs and agents that potentiate the activity of existing beta-lactams against low-affinity PBP-producing organisms. PMID- 15338053 TI - Angiogenesis and signal transduction in endothelial cells. AB - Endothelial cells receive multiple information from their environment that eventually leads them to progress along all the stages of the process of formation of new vessels. Angiogenic signals promote endothelial cell proliferation, increased resistance to apoptosis, changes in proteolytic balance, cytoskeletal reorganization, migration and, finally, differentiation and formation of a new vascular lumen. We aim to review herein the main signaling cascades that become activated in angiogenic endothelial cells as well as the opportunities of modulating angiogenesis through pharmacological interference with these signaling mechanisms. We will deal mainly with the mitogen-activated protein kinases pathway, which is very important in the transduction of proliferation signals; the phosphatidylinositol-3-kinase/protein kinase B signaling system, particularly essential for the survival of the angiogenic endothelium; the small GTPases involved in cytoskeletal reorganization and migration; and the kinases associated to focal adhesions which contribute to integrate the pathways from the two main sources of angiogenic signals, i.e. growth factors and the extracellular matrix. PMID- 15338054 TI - Role of nitric oxide in the functional response to ischemia-reperfusion of heart mitochondria from hyperthyroid rats. AB - We investigated the role of nitric oxide (NO) in the mitochondrial derangement associated with the functional response to ischemia-reperfusion of hyperthyroid rat hearts. Mitochondria were isolated at 3000 g from hearts subjected to ischemia-reperfusion, with or without N(omega)-nitro-L-arginine (L-NNA, an NO synthase inhibitor). During reperfusion, hyperthyroid hearts displayed tachycardia and low functional recovery. Their mitochondria exhibited O(2) consumption similar to euthyroid controls, while H(2)O(2) production, hydroperoxide, protein-bound carbonyl and nitrotyrosine levels, and susceptibility to swelling were higher. L-NNA blocked the reperfusion tachycardic response and increased inotropic recovery in hyperthyroid hearts. L-NNA decreased mitochondrial H(2)O(2) production and oxidative damage, and increased respiration and tolerance to swelling. Such effects were higher in hyperthyroid preparations. These results confirm the role of mitochondria in ischemia-reperfusion damage, and strongly suggest that NO overproduction is involved in the high mitochondrial dysfunction and the low recovery of hyperthyroid hearts from ischemia reperfusion. L-NNA also decreased protein content and cytochrome oxidase activity of a mitochondrial fraction isolated at 8000 g. This and previous results suggest that the above fraction contains, together with light mitochondria, damaged mitochondria coming from the heaviest fraction, which has the highest cytochrome oxidase activity and capacity to produce H(2)O(2). Therefore, we propose that the high mitochondrial susceptibility to swelling, favoring mitochondrial population purification from H(2)O(2)-overproducing mitochondria, limits hyperthyroid heart oxidative stress. PMID- 15338055 TI - Global gene expression of fission yeast in response to cisplatin. AB - The cellular response to the antitumor drug cisplatin is complex, and resistance is widespread. To gain insights into the global transcriptional response and mechanisms of resistance, we used microarrays to examine the fission yeast cell response to cisplatin. In two isogenic strains with differing drug sensitivity, cisplatin activated a stress response involving glutathione-S-transferase, heat shock, and recombinational repair genes. Genes required for proteasome-mediated protein degradation were up-regulated in the sensitive strain, whereas genes for DNA damage recognition/repair and for mitotic progression were induced in the resistant strain. The response to cisplatin overlaps in part with the responses to cadmium and the DNA-damaging agent methylmethane sulfonate. The different gene groups involved in the cellular response to cisplatin help the cells to tolerate and repair DNA damage and to overcome cell cycle blocks. These findings are discussed with respect to known cisplatin response pathways in human cells. PMID- 15338056 TI - Hcc-1 is a novel component of the nuclear matrix with growth inhibitory function. AB - Hcc-1 is a novel nuclear protein containing the SAF-box DNA-binding domain. It binds to both double-stranded and single-stranded DNA with higher affinity for the single-stranded form. In addition, it also binds specifically to scaffold/matrix attachment region DNA. These nucleic acid-binding characteristics suggest a potential function for Hcc-1 as a component of the heterogeneous ribonucleoprotein complex. Using a yeast two-hybrid screen, two DEAD-box RNA helicases, BAT1 and DDX39, were identified as proteins that interact with Hcc-1. Interactions with these RNA helicases suggested a role for Hcc-1 in nucleic acid biogenesis. Expression of Hcc-1 in the HEK293 cell line resulted in a slower growth rate compared to controls (p = 0.0173) and an accumulation of cells at the G2/M phase (p = 0.0276 compared to control HEK293 cells). Taken together, these results suggest a role for Hcc-1 in growth regulation and nucleic acids metabolism. PMID- 15338057 TI - Impact of neuropathic pain on the gene expression and activity of cytochrome P450side-chain-cleavage in sensory neural networks. AB - Development of efficient therapy against chronic and stubborn pains requires fundamental identification of adequate cellular and molecular targets. This study combined cellular, molecular and biochemical approaches to investigate the gene expression and enzymatic activity of cytochrome P450side-chain-cleavage (P450scc) in spinal neural networks under normal and neuropathic pain states. P450scc is the key onset enzyme for steroidogenesis in endocrine glands and for neurosteroid biosynthesis in nerve cells. The P450scc gene was over-expressed in spinal and supra-spinal networks during neuropathic pain provoked by sciatic nerve ligature. Plasticity was observed in P450scc cellular distribution in pain circuits and its activity also increased inducing in vivo, hyper-secretion of pregnenolone and allopregnanolone which strongly stimulates type A receptors for g-aminobutyric acid, a pivotal neurotransmitter involved in pain modulation. These results, by establishing a direct link between neuropathic pain and neuroactive steroid formation in the nervous system, open new perspectives for chronic-pain modulation by endogenous neurosteroids. PMID- 15338058 TI - A short history of theoretical surgery. PMID- 15338059 TI - Adverse reactions to drugs: in vitro studies with isolated cells. AB - OBJECTIVE AND DESIGN: Drug-induced adverse reactions can be allergic or pseudoallergic in nature, in this study the histamine releasing ability of 4 radiographic contrast media and 2 opioid analgesics was tested on a variety of mast cell containing cell suspensions. MATERIAL: Mast cell containing cell suspensions were obtained from porcine lung, liver, kidneys and heart, as well as rat lung and rat peritoneal lavage. TREATMENT: Cells were incubated for 10 min with Angiographin (amidotrizoate), Hexabrix (ioxaglate), Rayvist (ioglycate) or Telebrix (ioxithalamate) all 10-500 microl/ml or with levomethadone (0.1-5 mM) or pethidine (0-10 mM). METHODS: Histamine was measured using an automated fluorometric method and percentage histamine release calculated. RESULTS: All agents caused histamine release from porcine cells and rat lung cells. However, rat peritoneal mast cells were refractory to the action of pethidine. Cardiac cells were the most sensitive to the radiographic contrast media but the least sensitive to the opioid analgesics. CONCLUSIONS: The data indicate that mast cells isolated from animal models can provide an indication of those agents likely to induce an adverse pseudoallergic reaction. However, these data should be used together with those obtained from human mast cells, in vivo animal experiments as well as studies using human volunteers and clinical trials as advocated in the Marburg Model. PMID- 15338060 TI - Unexpected partial H1-receptor agonism of imidazole-type histamine H3-receptor antagonists lacking a basic side chain. AB - OBJECTIVE AND DESIGN: The putative partial H1-receptor agonism of some H3 receptor antagonists belonging to the proxifan series was characterized in a functional in-vitro assay using guinea-pig ileum. METHODS: Whole segments of guinea-pig ileum were mounted in Tyrode's solution under isotonic conditions in the presence of atropine (10(-7) M) and were cumulatively treated with histamine as an internal reference. After washout, the putative H1-receptor agonists were added cumulatively to determine agonist potency (pEC50) and intrinsic activity (Emax) relative to histamine. Maximal or supramaximal concentrations of partial agonists, or sufficient concentrations of H1-receptor antagonists were incubated for 3-15 min prior to construction of a second concentration-effect curve to histamine in order to calculate partial agonist or antagonist affinity for the H1 receptor (pKP or pA2 value, respectively). RESULTS: Several analogues of FUB 372 displayed low H1-receptor affinities (pA2 or pKP 4.2-5.5) except for a methyl benzoate derivative (pA2 = 6.81, Schild plot slope unity). FUB 372, four ortho substituted derivatives (R = F, CH3, OCH3, CF3), and ciproxifan were weak contractile agents (Emax 9-38%, pEC50 4.73-5.68, histamine: 6.70) susceptible to antagonism by the H1-antihistaminergic drug mepyramine (2.10(-9)-10(-7) M). Agonist potency and H1-receptor affinity of these compounds did not correlate with the data of a set of H1-histaminergic 2-phenylhistamines bearing the same substituents. CONCLUSIONS: A specific subset of proxifans related to FUB 372 and ciproxifan represent a unique type of H1-receptor agonists lacking a basic side chain. PMID- 15338061 TI - Antibiotic prophylaxis influences cardiovascular stability in complicated surgery. AB - OBJECTIVE: Antibiotic prophylaxis is used in many surgical procedures but there are frequent cardiovascular instabilities following antibiotics in perioperative period. A clinic modelling randomised trial (CMRT) in pigs was developed to compare the effects of 2 commonly used antibiotic combinations on cardiovascular stability during major surgery. MATERIALS AND METHODS: Thirty pigs (both sexes) were randomised into 3 groups, receiving either saline (placebo), co-amoxiclav or cefuroxime/metronidazole in clinically relevant doses as antibiotic prophylaxis. A laparotomy was performed and the abdomen remained open. Surgical complications were simulated by removing one third of the blood volume. For fluid resuscitation, 500 ml hetastarch (HAES(TM)) were infused rapidly (therapy of complication) and polymyxin B (15 mg/kg bodyweight) was applied for induction of histamine release reactions (complication of therapy). The main end points were histamine release reactions, these were classified by 2 blinded investigators. RESULTS: Neither cardiovascular changes nor histamine release reactions were detected immediately after the administration of antibiotics or placebo alone. Plasma histamine concentrations increased after bleeding in the co-amoxiclav group (p < 0.05). After fluid resuscitation and induction of anaphylactoid reactions, the median histamine release and cardiovascular changes were not significantly different between the groups. However, the incidence of typical histamine release related reactions differed significantly between the groups: 8/10 for the controls, 6/10 in the co-amoxiclav and 2/10 in the cefuroxime/metronidazole group (p < 0.05). CONCLUSIONS: The stability and reproducibility of this model clearly demonstrated the concept of a 'clinic modelling randomised trial' as a useful tool. Antibiotic prophylaxis influences the organism's capability to cope with intraoperative bleeding and fluid resuscitation problems. Indeed antibiotic prophylaxis may be beneficial. These effects of antibiotics could only be demonstrated in complex surgical models. Thus new antibiotics should be investigated in complex animal models prior to prospective randomised clinical trials or usage in clinical practice. PMID- 15338062 TI - Pre-treatment with ozonized oxygen (O3) aggravates inflammation in septic rats. AB - OBJECTIVE AND DESIGN: Ozone is produced by neutrophils during bacterial killing. Its application was found to be beneficial in peritonitis patients. Therefore, we measured survival and cytokines after ozone pre-treatment in septic rats. SUBJECTS AND TREATMENT: With approval, 40 male Wistar-rats were allocated to 1) ozone pre-treatment for five days before intra-abdominal sepsis, or 2) no pre treatment. METHODS: The primary endpoint was mortality at 120 h. Secondary endpoints were plasma cytokine levels. RESULTS: In the control group mortality was 50% (10/20 rats). After ozone pre-treatment, survival was only 35% (7/20 rats, Log-Rank test: P = 0.10). Ozone increased TNF-alpha and MIP-2 after infection: 127 +/- 23 pg/ml and 94 +/- 19 pg/ml (control group: 398 pg/ml and 369 pg/ml; P < 0.002 and P < 0.01). IL-6 levels were similar in both groups. CONCLUSIONS: Ozone pre-treatment was pro-inflammatory in sepsis with a trend to reduced survival. Therefore, its effects in sepsis should be further evaluated in animal trials. PMID- 15338063 TI - Effects of G-CSF and antibiotic prophylaxis in a 2 x 2 factorial design on outcome in septic rats. AB - OBJECTIVE: In a recently completed randomised clinical trial in patients with colorectal cancer resections the combination of the granulocyte-colony stimulating factor (G-CSF) + cefuroxime/ metronidazole (cef/met) was superior to ofloxacin/metronidazole (ofl/met). These combinations were used to confirm the clinical data and to validate the concept of clinic modelling randomised trials (CMRTs) in a rat model of intra-abdominal sepsis. SUBJECTS: 80 male Wistar rats were randomised in a 2 x 2 factorial study design. TREATMENT: All animals (n = 20/group) received anaesthesia, antihistamines, antibiotic prophylaxis, peritoneal contamination and infection. Groups were: 1) G-CSF + cef/met; 2) placebo + cef/ met; 3) G-CSF+ofl/met; 4) placebo + ofl/met. G-CSF (20 g/kg) prophylaxis was applied three times. METHODS: Survival at 120 h was analysed with the Kaplan Meier method. RESULTS: Survival rate was best in the G-CSF + cef/met group with 75% and was significantly improved compared to the cef/met placebo group, in which only 42% survived (P < 0.05). Survival rate between both G-CSF groups was similar being 75% in the cef/met and 72% in the ofl/met group. P = 0.10). Ozone increased TNF-alpha and MIP-2 after infection: 127 +/- 23 pg/ml and 94 +/- 19 pg/ml (control group: 398 pg/ml and 369 pg/ml; P < 0.002 and P < 0.01). IL-6 levels were similar in both groups. CONCLUSIONS: The results of this CMRT confirmed the result of our clinical G-CSF trial in that G-CSF prophylaxis was most efficacious in combination with cef/met to improve the outcome. PMID- 15338064 TI - Understanding quality of life in patients with colorectal cancer: comparison of data from a randomised controlled trial, a population based cohort study and the norm reference population. AB - BACKGROUND AND AIMS: Quality of life (QoL) is an important outcome measure in clinical studies. However, there is little experience with the interpretation of QoL results. METHODS: To guide interpretation of QoL results from a randomised controlled trial (RCT) targeting the effectiveness of the immune modulator G-CSF on postoperative recovery in high risk (ASA III/IV) colorectal cancer patients, we compared RCT data with data from a population based cohort study and norm reference data. QoL was assessed using the European Organisation for the Research and Treatment of Cancer (EORTC) QLQ-C30 and CR38 questionnaires. QoL results were analysed on discharge from hospital and six months postoperatively. RESULTS: Colorectal cancer patients (both from the RCT and the cohort study) showed the greatest differences in QoL scores compared to norm reference data at discharge from hospital. Six months postoperatively, global quality of life and pain approximated norm reference values indicating optimal recovery. However, deficits still appeared in scores for role functioning, physical functioning, social functioning and fatigue. The best improvements (discharge from hospital to six months postoperatively) were seen with respect to physical functioning, fatigue and pain. CONCLUSIONS: For further analysis of RCT data, physical functioning and fatigue scores may be more sensitive than global quality of life to detect differences in treatment effects. PMID- 15338065 TI - Survival chances and psychological aspects of quality of life in patients with localized early stage breast cancer. AB - BACKGROUND: The present analysis focuses on the long term psychological reactions to early stage breast cancer. Two hypotheses were formulated. The first hypothesis draws a direct link between tumour size/survival chances and Quality of life (QoL): The better the survival chances, the better QoL ('biological danger model'). The second hypothesis assumes that localized early breast cancer has excellent prognosis (> 90% five year survival rate), and that therefore QoL differences between various forms of early breast cancer should be minimal ('medico-pragmatic model'). PATIENTS AND METHODS: In a defined rural area with 252.000 inhabitants (small-area-analysis), a total of n = 389 patients with primary breast cancer were recruited. For the present analysis we selected a subgroup (n = 269) from the cohort by tumour size (pTis, pT1a,b, pT1c, and pT2). QoL scores for global quality of life, emotional functioning and future perspective were computed according to the EORTC manual and compared to age matched norm data of the German population. RESULTS: A total of 690 QoL questionnaires were obtained from n = 269 patients with comparable completion rates within the four subgroups (pTis, pT1a,b, pT1c, and pT2). For all four groups and in all scores there were improvements over time. Generally, pTis always scored highest, pT2 always lowest, the other two groups in between. After one year pTis patients had higher mean scores in global quality of life than the norm. In contrast, pT1a,b were considerably lower than the norm and the difference between these two was 17.2 score points. It seems that the small difference (3.5%) in five year survival chances between pTis and T1 a,b tumours transforms into marked differences regarding quality of life, thus supporting a biological danger model of the survival/QoL relationship. CONCLUSIONS: Our results show that physicians have to realise although their early breast cancer patients have excellent survival chances, psychological distress is present. From a clinical perspective we would recommend that early stage breast cancer patients, and especially patients with occult, pT1a,b tumours be informed about their excellent prognosis. In addition, cognitive therapy might help patients stop worrying about their cancer. PMID- 15338066 TI - Attitude towards clinical trials: results of a survey of persons interested in research. AB - OBJECTIVE AND DESIGN: Patient recruitment is a major problem in clinical trials. In a survey, the attitude of a public sample towards clinical trials was assessed. SUBJECTS: In a survey 225 visitors to the Heinrich-Heine University were interviewed with respect to their attitude towards clinical trials. METHODS: Visitors were interviewed with regard to sociodemographic variables, disease status, knowledge about and experience of trials and attitude towards clinical trials. Three scenarios for clinical trials were presented (dental trial, surgical trial, pharmaceutical trial). RESULTS: In general, a positive attitude was found. Clinical trials were judged important by 89.5% of the survey participants; however, only 25% would take part in clinical trials. The likelihood of potential participation was lower in the surgical trial than in the dental or pharmaceutical trial scenarios. The willingness to participate was significantly higher in persons, who considered trials to be important, with general knowledge about clinical trials and with previous trial participation. CONCLUSIONS: The willingness to participate in a clinical trial is still low in the public. More information and involvement in clinical trials may improve the general attitude. PMID- 15338067 TI - Influence of histamine release on postoperative vomiting (POV) following gynaecological laparoscopic surgery. AB - OBJECTIVE: The mechanisms leading to the high incidence of postoperative vomiting (POV) after gynaecological laparoscopic surgery are still unknown. The effectiveness of POV-prophylaxis using H1 + H2-receptor antagonists has been demonstrated, suggesting a role for histamine in the pathogenesis of POV. However, histamine levels were not measured in these studies. The aim of this study was to investigate the incidence of plasma histamine release and its association with POV after gynaecological laparoscopic surgery. MATERIAL OR SUBJECTS: Twenty-two female patients, aged 20-56 y, classified ASA physical status I or II, undergoing elective gynaecological laparoscopic surgery were enrolled in the study. Blood samples for plasma histamine measurements were drawn at defined time points perioperatively. Emetic symptoms were recorded within the first 24 h after operation. A standardized balanced anaesthesia without any prophylactic antiemetic medication was applied. Formal causality analysis for histamine as a determinant for POV was performed. RESULTS: The overall incidence of POV was 40.9% (9 out of 22 patients). Twelve out of 22 patients (54.5%) demonstrated a histamine release reaction during the whole observation period. Six out of 9 patients with POV (66.7%) had a histamine release. There was no difference in mean plasma histamine levels between POV-positive and POV-negative patients. The conditional probability for POV with histamine release was 6/12 = 0.5, in contrast to 3/10 = 0.3 for POV without histamine release. CONCLUSIONS: A high incidence of plasma histamine release was demonstrated in most but not all patients with POV. The probability of POV with histamine release (0.5) was higher than without histamine release (0.3), thus histamine release was shown to be one of the contributory determinants for POV in this clinical study. Thus, patients at risk for POV may benefit from a H1 + H2-receptor antagonists prophylaxis alone or in combination with other antiemetic strategies. PMID- 15338068 TI - Premedication with H1 and H2 blocking agents reduces the incidence of postoperative nausea and vomiting. AB - OBJECTIVE: Patients undergoing anaesthesia and surgery frequently complain about postoperative nausea and vomiting (PONV). Whether pretreatment with H1 and H2 blocking agents reduces the incidence of PONV remains controversial. To answer this question, we performed a randomised, prospective, placebo-controlled clinical study to evaluate the efficacy of a premedication with H1 and H2 receptor antagonists. MATERIAL AND SUBJECTS: 1149 patients (both sexes) undergoing surgery were randomly assigned to three treatment groups and one control group. Patients in the treatment groups were premedicated with the following H1 + H2 receptor antagonists:Group 1 (n = 335): 5 mg/kg cimetidine i.v. + 0.1 mg/kg dimetindene i.v. 20 min before induction of anaesthesiaGroup 2 (n = 337): 1.25 mg/kg ranitidine i.v. + 0.1 mg/kg dimetindene i.v. 20 min before induction of anaesthesiaGroup 3 (n = 316): 300 mg ranitidine p.o. + 0.1 mg/kg dimetindene i.v. 1 to 2 h before induction of anaesthesiaGroup 4 (n = 161): 20 ml saline solution i.v. 20 min before induction of anaesthesiaPatients from the treatment groups 1, 2 and 3 received regional or general anaesthesia depending on the clinical decision. All control patients received general anaesthesia consisting of fentanyl, a thiobarbiturate, enflurane, nitrous oxide, oxygen, and vecuronium. RESULTS: The incidence of nausea and vomiting was 8.5%, 6.8% and 5.4% in patients from the treatment groups (1, 2 and 3) who underwent general anaesthesia (n = 545), with no statistically significant differences between groups. The incidence of nausea and vomiting in the control group (n = 161) was 28.3% (nausea) and 27.5% (vomiting), respectively. In patients who underwent regional anaesthesia (n = 443), the incidence of nausea and vomiting was 2.5% and 1.1%, respectively. CONCLUSIONS: Premedication with H1 and H2 blocking agents significantly reduces the incidence of postoperative nausea and vomiting. PMID- 15338069 TI - The effect of Palamed G bone cement on early migration of tibial components in total knee arthroplasty. AB - OBJECTIVE: Migration of the tibial component in total knee arthroplasty (TKA) is subject of many studies using roentgen stereophotogrammetric analysis (RSA). In previous studies of cemented and uncemented tibial components, high migration values were found. Improvements in cementing technique, prosthetic design and pre coating techniques reduced these values as shown in more recent studies. MATERIAL AND SUBJECTS: A total of 35 patients were initially included in the study and operated on between 12/1999 and 10/2000. All patients received a NexGen TKA cemented into the proximal tibia using Palamed G bone cement. The implants and the tibial metaphysis were marked with standard tantalum markers. Radiostereometric analysis was performed post-operatively and after 3, 6 and 12 months using a standard digital radiostereometric analysis. Functional parameters were assessed using the Knee Society Score (KSS) clinical rating system. RESULTS: There were no complications and failures within the first year. After 1 year radiostereometric measurements of the translational parameters along and the rotational parameters around the x-, y- and z-axis revealed: X-Trans -0.19 mm, Y Trans +0.02 mm, Z-Trans +0.08 mm, X-Rot +0.26 degrees, Y-Rot -0.35 degrees, Z-Rot +0.09 degrees. The maximum total point motion was +0.96 mm and the mean maximum subsidence was -0.23 mm. Except for anterior-posterior, medio-lateral stability and extension leg all endpoints of the KSS clinical rating system showed a significant improvement. CONCLUSIONS: After 12 months, the use of Palamed G bone cement in total knee arthroplasty was demonstrated to be safe. Both the clinical and radiostereometric results were good and comparable to the results reported in other RSA studies in cemented total knee arthroplasty. PMID- 15338070 TI - Classification of perioperative histamine-related reactions. AB - OBJECTIVE: Histamine release may cause anaphylactoid reactions. However, during anaesthesia and surgery especially cardiovascular effects may not be regarded as histamine-related. Therefore, we adapted the classical concept of histamine release reactions to the perioperative situation and validated the new paradigm. METHODS: Elevated plasma histamine (diagnostic gold standard) was correlated to potentially related intraoperative signs and symptoms. The validity, repeatability and sensitivity of the 'gold standard' was tested by ROC analysis in volunteers, who also received H1-/H2-histamine antagonists. Additionally, a dose-response relationship was determined in dogs using the histamine releaser compound 48/80. RESULTS: The 'gold standard' had a sensitivity of 96% (90%-100%) and a specificity of 93% (85%-100%). The reproducibility was proven by repeated injections of histamine. Skin reactions, tachycardia and hypertension were identified as histamine-related diagnostic variables. A dose-response curve of plasma histamine release was created. CONCLUSIONS: The defined 'gold standard' is valid for the diagnosis of histamine-related reactions during anaesthesia and surgery. It may help to identify patients, who could benefit from pre-anaesthetic antihistamine prophylaxis. PMID- 15338071 TI - Limitations of score-based daily outcome predictions in the individual intensive care patient. An example of the RIAHDH algorithm. AB - OBJECTIVE: The nature of score-based predictions is probabilistic, and their accuracy depends on the reliability and validity of the applied system. As an example, the present study investigates the accuracy of the RIP-algorithm (RIP = Riyadh Intensive Care Program) based on daily APACHE II scores, and compares it with published results of that algorithm from other investigators. DESIGN: Prospective observational study and review of the literature. PATIENTS AND METHODS: 1,986 consecutive admissions of 1,808 patients to a surgical intensive care unit were documented. Daily changes of score values were used to derive a risk of death estimation. Sensitivity and the rate of false predictions were calculated for score-based predictions. Health status one year after discharge was assessed in survivors predicted to die. RESULTS: Daily application of the algorithm identified 109 situations leading to death predictions in 56 patients. Five of these patients were discharged alive from the hospital (positive predictive value 91%). One year later 3 of these patients were still alive. The algorithm identified 51 of the non-survivors (sensitivity 19%), 110 died in the ICU without prediction. Altogether 270 patients died during their hospital stay. Among the 6 independent validation studies, similar results were found, but differences occurred due to the problematic assessment of consciousness. CONCLUSIONS: Sequential assessment of scores in intensive care could identify high risk patients, but with some degree of uncertainty. Therefore, the scores should only be used by those familiar with their limitations and risks. PMID- 15338072 TI - Computerized visualization of quality of life data of individual cancer patients- the QoL-Profiler. AB - OBJECTIVE: Quality of life (QoL) is increasingly considered as an important endpoint in clinical studies but difficult to use in clinical practice. For daily clinical practice, we developed a computer program that is able to calculate and draw QoL profiles for individual cancer patients. METHODS: The computer program was developed in several steps during the course of studies with different patient populations (prospective cohort study, randomised surgical trial, breast cancer patients, all tumour patients of a clinic) and using different software packages. RESULTS: The current version is based on Microsoft ACCESS and combines QoL data and medical data. Automated QoL profile output comprises 10 scores that are of clinical relevance. Scores range from 0 (worst) to 100 (best), with 50 considered as the threshold for intervention. CONCLUSIONS: Practitioners found QoL-profiles comprehensible and clinically useful. QoL profiles are the crucial link between the QoL concept and QoL enhancing treatment decisions. PMID- 15338073 TI - A problem solving and decision making toolbox for approaching clinical problems and decisions. AB - In this paper, we begin by presenting three real patients and then review all the practical conceptual tools that have been suggested for systematically analyzing clinical problems. Each of these conceptual tools (e.g. Evidence-Based Medicine, Clinical Practice Guidelines, Decision Analysis) deals mainly with a different type or aspect of clinical problems. We suggest that all of these conceptual tools can be thought of as belonging in the clinician's toolbox for solving clinical problems and making clinical decisions. A heuristic for guiding the clinician in using the tools is proposed. The heuristic is then used to analyze management of the three patients presented at the outset. PMID- 15338074 TI - On the origins and development of evidence-based medicine and medical decision making. AB - The aims of this paper are to identify the issues and forces that were the impetus for two recent developments in academic medicine, evidence-based medicine (EBM) and medical decision making (MDM); to make explicit their underlying similarities and differences; and to relate them to the fates of these innovations. Both developments respond to concerns about practice variation; the rapid growth of medical technology, leading to a proliferation of diagnostic and treatment options; the patient empowerment movement; and psychological research that raised questions about the quality of human judgment and decision making. Their commonalities include: use of Bayesian principles in diagnostic reasoning, and the common structure embedded in an answerable clinical question and a decision tree. Major differences include: emphasis on knowledge or judgment as the fundamental problem; the status of formal models and utility assessment; and the spirit and tone of the innovation. These differences have led to broader acceptance of EBM within academic medicine, while decision analysis, the fundamental tool of MDM, has been less welcomed in clinical circles and has found its place in guideline development, cost-effectiveness analysis, and health policy. PMID- 15338075 TI - Wilfred Lorenz--the theoretical surgeon. How it started. PMID- 15338076 TI - Qualified practice guidelines--advantage or restriction of medical health care? PMID- 15338077 TI - Perspectives of surgical research. PMID- 15338078 TI - Purification and characterization of NADPH-dependent aldo-keto reductase specific for beta-keto esters from Penicillium citrinum, and production of methyl (S)-4 bromo-3-hydroxybutyrate. AB - A novel beta-keto ester reductase (KER) was purified to homogeneity from recombinant Escherichia coli (pTrcKER) cells, which efficiently expressed the ker gene cloned from Penicillium citrinum IFO4631. The enzyme was monomeric and had a molecular mass of 37 kDa. It catalyzed the reduction of some beta-keto esters, especially alkyl 4-halo-3-oxobutyrates. However, it did not catalyze the reverse reaction, the dehydrogenation of alkyl 4-halo-3-hydroxybutyrates and other alcohols. The enzyme required NADPH as a cofactor and showed no activity with NADH. Therefore, it was defined as a NADPH-dependent aldo-keto reductase (AKR3E1), belonging to the AKR superfamily. The enzyme stereospecifically produced methyl (S)-4-bromo-3-hydroxybutyrate from its keto derivative with high stereospecificity (97.9% enantiomer excess). E. coli cells expressing KER and glucose dehydrogenase in the water/butyl acetate two-phase system achieved a high productivity of (S)-4-bromo-3-hydroxybutyrate (277 mM, 54 mg/ml) in the organic solvent layer. PMID- 15338079 TI - Production of bacterial cellulose by Acetobacter xylinum BPR2001 using molasses medium in a jar fermentor. AB - Bacterial cellulose (BC) production by Acetobacter xylinum subsp. sucrofermentans BPR2001 using molasses medium was carried out in a jar fermentor. When molasses was subjected to H(2)SO(4)-heat treatment, the maximum BC concentration increased to 76% more than that achieved using untreated molasses, and the specific growth rate increased 2-fold. When the initial sugar concentrations in the H(2)SO(4) heat treated molasses were varied from 23 g/l to 72 g/l, BC concentration, production rate, and yield were maximum at sugar concentrations of 23 g/l and 37 g/l, and production of by-products, such as polysaccharides and CO(2), was lower than at sugar concentrations of 48 g/l and 72 g/l, indicating that maintaining a lower molasses concentration is essential for efficient BC production in jar fermentors, this being due mainly to the complex nature of molasses. Molasses has a clear advantage over pure sugars as a carbon source from an economic viewpoint. PMID- 15338080 TI - Robust NADH-regenerator: improved alpha-haloketone-resistant formate dehydrogenase. AB - Formate dehydrogenases (FDH) are useful for the regeneration of NADH, which is required for asymmetric reduction by several dehydrogenases and reductases. FDHs have relatively low activity and are labile, especially to alpha-haloketones, thus FDH cannot be applied to the industrial manufacture of optically active alpha-haloalcohols. To stabilize a FDH from Mycobacterium vaccae (McFDH) against the alpha-haloketone ethyl 4-chloroacetoacetate (ECAA), a set of cysteine-mutant enzymes was constructed. Sensitivity to ECAA of mutant C6S was similar to that of the wild-type enzyme, and mutants C249S and C355S showed little activity. In contrast, mutant C256S exhibited remarkable tolerance to ECAA. Surprisingly, mutant C146S was activated by several organic compounds such as ethyl acetate. An optimized mutant, C6A/C146S/C256V (McFDH-26), was obtained by combining several effective mutations. Ethyl (S)-4-chloro-3-hydroxybutanoate [(S)-ECHB] was synthesized from ECAA to 49.9 g/l with an optical purity of more than 99% e.e. using recombinant Escherichia coli cells coexpressing McFDH-26 and a carbonyl reductase (KaCR1) from Kluyveromyces aestuarii. PMID- 15338081 TI - Fugu immunoglobulin D: a highly unusual gene with unprecedented duplications in its constant region. AB - We have isolated and characterized the cDNA that encodes IgD of fugu (Takifugu rubripes). Though the splicing of micro1 with the delta1 domain was similar to those reported for teleost IgDs, highly unusual and unprecedented domain duplications were found in the constant region of the fugu IgD. The structure of the fugu IgD is like VDJ-micro1-(delta1-delta2-delta3-delta4-delta5-delta6)(2) delta7-deltam1-deltam2. Genomic sequence analysis of the fugu IgD gene supported the results of cDNA sequencing that the first six delta domains in the constant region are duplicated. Such a novel duplication pattern has not been reported in any other vertebrates. However, IgD secretory domains could not be identified in this study. The deduced amino acid sequence of the fugu IgD constant region showed high identity (35-55%) to the sequences of previously reported teleost IgDs. Gene expression analyses based on RT-PCR demonstrated that the IgD gene is preferentially expressed in presumptive lymphoid tissues; moreover, in situ hybridization showed that IgD-positive cells are distributed throughout the spleen and head kidney. The expression pattern is similar to that of IgM, corroborating the hypothesis that IgD plays an important role in the humoral immune system of this species. PMID- 15338082 TI - Transcranial Doppler imaging in children: sickle cell screening and beyond. AB - Transcranial Doppler (TCD) is widely accepted as the modality of choice for screening intracranial vessels in children with sickle cell disease. Its advantages are that it is noninvasive (no need for sedation, contrast material, or radiation), portable, easily repeated, and it provides information about the intracranial vessels that is otherwise unavailable. These positive attributes explain why in recent years the applications for TCD have grown beyond sickle cell screening to almost any disease process that involves the major intracranial vessels. The objective of this manuscript is to discuss key points on how to perform and interpret TCD, and discuss imaging features of various pathological processes such as sickle cell, asphyxia, brain trauma, brain death, hydrocephalus, enlarged subarachnoid spaces, vasospasm, vasculitis, venous sinus thrombosis, and vein of Galen malformation. PMID- 15338083 TI - Kniest dysplasia: MR correlation of histologic and radiographic peculiarities. AB - Unossified epiphyses of Kniest dysplasia patients histologically reveal a bizarre pattern of chondrocytes lying amid a highly vacuolated matrix giving rise to the name "Swiss cheese" cartilage. The ossified epiphyses also are unusual in showing clouds of dense punctate calcifications randomly distributed throughout. Both unossified and ossified epiphyses reveal on MR imaging a similar pattern of lakes of bright T2 signal against a relatively normal background, which represents an interesting analogue to the histologic and radiographic features. PMID- 15338085 TI - Abstracts of the 29th Annual Conference of the European Society of Neuroradiology and 13th Advanced Course. September 2004. Aachen, Germany. PMID- 15338084 TI - The effect of sustained and local administration of epidermal growth factor on improving bilateral testicular tissue after torsion. AB - Epidermal growth factor (EGF) modulates Leydig cell proliferation, steroidogenesis, spermiogenesis, and Sertoli cell activity. It plays an important role in repairing ischemia-reperfusion injury in different tissues. The aim of this study was to evaluate the effects of sustained and local administration of EGF on improving bilateral testicular tissue after torsion. A total of 57 Wistar albino rats were used. For the EGF transport system, 1x2 cm gelatin films containing 2 microg EGF were used. Torsion was created by rotating the right testis 720 degrees in a clockwise direction for 4 h in all groups except the control group. Then, in the torsion group, bilateral orchiectomy was performed. After returning the torsioned ipsilateral testes to their normal state, the bilateral testes were wrapped by 1x2 cm unloaded gelatin films in the gelatin (G7 and G21) groups and, by 2 microg EGF loaded gelatin films in the EGF 7 and EGF 21 groups. The testes were removed on the seventh and 21st days, respectively, for biochemical and histological examination. Histologically, Johnsen's spermatogenesis criteria and mean seminiferous tubule diameter (MSTD) measurements were used. The EGF7 group did not show significant loss of Sertoli cells, while in the G7 group the number of these cells decreased. The ipsilateral ischemic testis of the EGF21 group showed Leydig cell hyperplasia, and the contralateral non-ischemic testes in this group were similar to the control group. In the G21 group, the bilateral testes showed Sertoli cell only syndrome in some sections, and most of the cells were undergoing apoptosis. The mean spermatogenesis scores and MSTD in the EGF7 and EGF21 groups were higher than in the G7 and G21 groups ( P<0.05). Malondialdehyde levels were significantly lower in the EGF groups than in the G groups ( P<0.05). Glutathione peroxidase (GSH-Px) levels in the G21 group were significantly higher than in the EGF21 group. Our study shows that local and sustained EGF release after testicular torsion improves bilateral testicular injury. EGF administration may be a new treatment choice for bilaterally injured testis after detorsion without removing the twisted testis. PMID- 15338086 TI - Patterns and correlates of benzodiazepine use in the French general population. AB - OBJECTIVE: To investigate the prevalence of current use of benzodiazepines (BZDs) and related drugs in the French general population and factors associated with this use. METHODS: National cross-sectional telephone survey conducted between 25 April 2001 and 8 May 2001 in a representative sample of non-institutionalized adults of BZD use and duration, prescriber specialty, socio-demographic data and mood and anxiety disorders, using a structured diagnostic interview. RESULTS: The prevalence of current use of BZD was 7.5%. It was higher among women (9.7%) than men (5.2%). It increased with age and was higher in the jobless (10.9). Duration of BZD use was more than 6 months in 75.9% of users and increased with age. Of the 711 (17.7%) subjects with at least one mood or anxiety disorder, 122 (17%) used BZD compared with 180 (5.5%) of the 3296 subjects without mood or anxiety disorders. In multivariate analysis, factors associated with BZD use were age [odds ratio (OR): 3.6; 95% confidence interval (CI) 2.0-5.6], 6.5 (4.1-10.3) and 10.9 (6.9-17.1), respectively, for ages 35-44 years, 45-59 years and over 60 years compared with below 34 years, female gender (OR: 1.7; 95% CI 1.3-2.1), anxiety only (OR: 2.2; 95% CI 1.5-3.2), mood disorder only (OR: 4.4; 95% CI 2.7 7.1) or both mood and anxiety disorders (OR: 8.8; 95% CI 5.9-12.6). CONCLUSION: Despite precautions, warnings and attempts to limit use, there remains a high proportion of long-term BZD users in the general French population, especially in the elderly. Our findings add to the weight of opinion that messages concerning proper use of BZDs certainly need to be clarified and amplified. PMID- 15338087 TI - Closed head injury and perceptual processing in dual-task situations. AB - Using a classical psychological refractory period (PRP) paradigm we investigated whether increased interference between dual-task input processes is one possible source of dual-task deficits in patients with closed-head injury (CHI). Patients and age-matched controls were asked to give speeded motor reactions to an auditory and a visual stimulus. The perceptual difficulty of the visual stimulus was manipulated by varying its intensity. The results of Experiment 1 showed that CHI patients suffer from increased interference between dual-task input processes, which is related to the salience of the visual stimulus. A second experiment indicated that this input interference may be specific to brain damage following CHI. It is not evident in other groups of neurological patients like Parkinson's disease patients. We conclude that the non-interfering processing of input stages in dual-tasks requires cognitive control. A decline in the control of input processes should be considered as one source of dual-task deficits in CHI patients. PMID- 15338088 TI - A unified view of propagating and localized surface plasmon resonance biosensors. AB - The intense colors of noble metal nanoparticles have inspired artists and fascinated scientists for hundreds of years. In this review, we describe refractive index sensing platforms based on the tunability of the localized surface plasmon resonance (LSPR) of arrays of silver nanoparticles and of single nanoparticles. Specifically, the color associated with single nanoparticles and surface-confined nanoparticle arrays will be shown to be tunable and useful as platforms for chemical and biological sensing. Finally, the LSPR nanosensor will be compared to traditional, flat surface, propagating surface plasmon resonance sensors. PMID- 15338089 TI - Cantilever-based biosensors. AB - This review provides a general introduction into the theory of cantilever sensors, including practical design concepts, examples for liquid-phase sensing with particular emphasis on biological applications, and a discussion of future prospects. PMID- 15338090 TI - Biomolecular and amphiphilic films probed by surface sensitive X-ray and neutron scattering. AB - In this review article we discuss the thin film analytical techniques of interface sensitive X-ray and neutron scattering applied to aligned stacks of amphiphilic bilayers, in particular phospholipid membranes in the fluid L(alpha) phase. We briefly discuss how the structure, composition, fluctuations and interactions in lipid or synthetic membranes can be studied by modern surface sensitive scattering techniques, using X-rays or neutrons as a probe. These techniques offer an in-situ approach to study lipid bilayer systems in different environments over length scales extending from micrometer to nanometer, both with and without additional membrane-active molecules such as amphiphilic peptides or membrane proteins. PMID- 15338091 TI - Improvement of sensitivity in the determination of organochlorine pesticides using a PSS injector with GC-ECD. AB - This paper describes the optimisation of a PSS injector in a gas chromatograph with a programmed pneumatic control (PPC) for the determination of 21 organochlorine pesticides. The injection of high volumes of sample (20 microl) improves the detection limits and allows a reduction in the amount of sample processed. The injection conditions were selected by a Plackett-Burman design followed by a central composite design. The LODs obtained in the optimum conditions were compared with those obtained with splitless/ECD. Finally, the method was successfully applied to the analysis of a leachate and vegetable samples. PMID- 15338092 TI - Determination of diethanolamine or N-methyldiethanolamine in high ammonium concentration matrices by capillary electrophoresis with indirect UV detection: application to the analysis of refinery process waters. AB - Alkanolamines such as diethanolamine (DEA) and N-methyldiethanolamine (MDEA) are used in desulfurization processes in crude oil refineries. These compounds may be found in process waters following an accidental contamination. The analysis of alkanolamines in refinery process waters is very difficult due to the high ammonium concentration of the samples. This paper describes a method for the determination of DEA in high ammonium concentration refinery process waters by using capillary electrophoresis (CE) with indirect UV detection. The same method can be used for the determination of MDEA. Best results were achieved with a background electrolyte (BGE) comprising 10 mM histidine adjusted to pH 5.0 with acetic acid. The development of this electrolyte and the analytical performances are discussed. The quantification was performed by using internal standardization, by which triethanolamine (TEA) was used as internal standard. A matrix effect due to the high ammonium content has been highlighted and standard addition was therefore used. The developed method was characterized in terms of repeatability of migration times and corrected peak areas, linearity, and accuracy. Limits of detection (LODs) and quantification (LOQs) obtained were 0.2 and 0.7 ppm, respectively. The CE method was applied to the determination of DEA or MDEA in refinery process waters spiked with known amounts of analytes and it gave excellent results, since uncertainties obtained were 8 and 5%, respectively. PMID- 15338093 TI - Liquid chromatography coordination ion-spray mass spectrometry (LC-CIS-MS) of docosahexaenoate ester hydroperoxides. PMID- 15338094 TI - Preconcentration technique for nonylphenol using cellulose cotton with homogenous liquid-liquid extraction for liquid chromatographic analysis. AB - A powerful preconcentration method for nonylphenol (NP) has been developed for liquid-chromatography by combining the use of cellulose cotton (solid-phase extraction) with homogeneous liquid-liquid extraction. A 100 ml of sample solution was preconcentrated using cotton, and the eluate obtained (acetonitrile; 5 ml) was further preconcentrated to 50 microl within 10 min using a homogeneous liquid-liquid extraction method (volume ratio, 2,000-fold; 100 ml--> 50 microl). The sample concentration increases from preconcentration was 1,599-fold, and NP was extracted into the sedimented phase at 80%. The proposed method was applied to high performance liquid chromatography with fluorescence detection (FL/HPLC); the lowest determination limit obtained was 1.0 x 10(-9) mol l(-1). PMID- 15338095 TI - Development of a fluorescent microplate assay for determining cyanovirin-N levels in plasma. AB - A sensitive immunosorbent competition assay was developed for quantitation of the anti-HIV protein cyanovirin-N (CV-N) in plasma using a 96-well plate format and a fluorescent endpoint. The assay is based on the binding of CV-N in plasma to plate-bound anti-CV-N antibodies, followed by removal of the plasma and addition of europium-labeled CV-N (Eu3+ -CV-N) to compete for the remaining antibody sites. Detection by addition of a dissociative fluorescence enhancement solution and time-resolved fluorescence measurements allowed correlation to the concentration of the native CV-N in plasma. A linear detection range of 1-100 nM (r2>0.99) was obtained for CV-N in mouse plasma. This assay was then utilized for analysis of plasma levels of CV-N samples following subcutaneous injection of CV N into mice. The results of these studies confirmed the reliability and sensitivity of this assay and the feasibility of its use for pharmacokinetic studies in a variety of species. PMID- 15338096 TI - Rapid determination of mono and dinitrophenols by DPP, in the presence of lead and cadmium and using concentrated CaCl2 electrolyte. AB - The contamination of drinking water and industrial wastewaters is a critical environmental problem. The nitrophenol, dinitrophenol, cadmium, and lead contaminants are classified as hazardous compounds. Their rapid determination may be obtained using differential pulse polarography with concentrated electrolyte. CaCl2, which is very soluble to levels exceeding 5 mol l(-1), allows separation of coalescent peaks at 0.1 mol l(-1). A systematic study undertaken from 0.1 to 5 mol l(-1) shows good separation of lead and cadmium from the organic compounds, and optimization of the electrolyte concentration according to the objective is described. Preconcentration of real samples is necessary because pollution levels are usually very low. PMID- 15338099 TI - Studies on the role of dopamine D1 receptors in the development and expression of MDMA-induced behavioral sensitization in rats. AB - RATIONALE: There is a large body of evidence indicating that the mesoaccumbens dopamine pathway is critically involved in the expression of behavioral sensitization to amphetamine and cocaine, but its role in the development of sensitization to psychostimulants is not that sound. Very few studies, however, have examined the role of dopamine transmission in 3,4 methylenedioxymethamphetamine (MDMA)-induced sensitization. OBJECTIVES: The effects of the D1 receptor antagonist SCH 23390 on the development and expression of MDMA-induced behavioral sensitization were investigated in rats. METHODS: During the development phase of sensitization, SCH 23390 was administered 15 min before every administration of MDMA. After 12 days of withdrawal, a MDMA challenge dose was given and locomotor activity was measured. In separate experiments, 15 min before the challenge injection of MDMA, SCH 23390 was administered either systemically or directly into the core of the nucleus accumbens (NAc) of MDMA-pretreated rats. RESULTS: SCH 23390 did not prevent the development of MDMA-induced behavioral sensitization but completely blocked the expression when given before the challenge dose of MDMA. The same results were obtained when SCH 23390 was locally applied into the core of the NAc. CONCLUSIONS: The present data suggest that D1 receptor stimulation is not critical for the development of long-term MDMA sensitization, in agreement with what has been reported for cocaine. By contrast, expression of sensitization depends on the activation of D1 receptors located in the NAc core. PMID- 15338100 TI - Cortisol response to diazepam: its relationship to age, dose, duration of treatment, and presence of generalized anxiety disorder. AB - OBJECTIVE: Acute diazepam administration has been shown to decrease plasma cortisol levels consistent with decreased activity of the hypothalamic-pituitary adrenal axis, especially in individuals experiencing stress. However, the effects of chronic diazepam treatment on cortisol have been less studied, and the relationship to age, anxiety, duration of treatment, and dose are not well understood. METHOD: This double-blind placebo-controlled study examined acute and chronic effects of diazepam on plasma cortisol levels in young (19-35 years) and elderly (60-79 years) individuals with and without generalized anxiety disorder (GAD). Subjects received single oral challenges of placebo or diazepam (2.5 mg or 10 mg) in a placebo-controlled cross-over design, followed by 3 weeks of chronic daily treatment with 2.5 mg or 10 mg diazepam or placebo taken at 10 p.m., and then by a final acute challenge with a single oral dose of the same study medication received during chronic treatment. RESULTS: The elderly experienced significant reductions in plasma cortisol levels compared to placebo both in the initial challenge and during chronic treatment, but the young did not. However, cortisol response to drug was comparable in both groups. Final challenge did not produce any significant cortisol effects in either group and the cortisol response in the elderly was significantly reduced compared to the initial challenge. GAD status was not a factor in plasma cortisol responses to diazepam. CONCLUSIONS: Diazepam reduced cortisol both acutely and during chronic treatment, but not during final challenge, consistent with some tolerance development. This effect was most apparent in the elderly compared with the young adults and was not modulated by GAD status or dosage, and was not related to drug effects on performance and on self-ratings of sedation and tension. PMID- 15338101 TI - Effects of acute treatment with antidepressant drugs on sensorimotor gating deficits in rats. AB - RATIONALE: Schizophrenic patients have a deficit in prepulse inhibition (PPI) which can be modelled in rats by administration of direct or indirect dopamine (DA) receptor agonists and N-methyl-D-aspartate (NMDA) receptor antagonists. Moreover, antipsychotics reverse the disruptive effect of DA agonists and NMDA receptor antagonists in this rat model. Consequently, this model is considered as predictive of antipsychotic action in the clinic. However, the effect of compounds, such as antidepressants, used for other psychiatric disorders but also administered to patients with schizophrenia has not been well investigated in this model. Antidepressants have been suggested not to affect PPI in humans. Thus, antidepressants are not expected to antagonise PPI disruption in rats, and should normally be used as negative controls in this model. OBJECTIVES: To investigate the effects of three antidepressant compounds, a serotonin reuptake inhibitor, a dopamine reuptake inhibitor, and a noradrenaline reuptake inhibitor in the rat PPI model. METHODS: The effect of acute treatment with citalopram, bupropion and desipramine on d-amphetamine-disrupted and phencyclidine (PCP) disrupted PPI in rats was investigated. Ziprasidone was tested as a positive control. RESULTS: None of the antidepressants, in contrast to ziprasidone, reversed PCP-disrupted PPI in rats. Both desipramine and ziprasidone normalised d amphetamine-disrupted PPI, while citalopram and bupropion were inactive. CONCLUSIONS: PCP-disrupted PPI in rats was less sensitive to false positives than the d-amphetamine-disrupted PPI model, based on the antidepressants tested in this study. PMID- 15338102 TI - Escalation of intravenous cocaine self-administration, progressive-ratio performance, and reinstatement in rats selectively bred for high (HiS) and low (LoS) saccharin intake. AB - RATIONALE: Rats selectively bred for high saccharin (HiS) intake consume more alcohol and acquire intravenous (IV) cocaine self-administration more rapidly than their low saccharin (LoS) consuming counterparts. OBJECTIVES: The purpose of the present study was to determine whether HiS and LoS rats also differ in the escalation, maintenance, and reinstatement of IV cocaine self-administration. METHODS: LoS and HiS female rats were allowed to self-administer cocaine [0.4 mg/kg; fixed ratio (FR) 1] under short (ShA, 2 h per day) or long (LgA, 12 h per day) access conditions for 21 days. Session lengths were subsequently equated (2 h) and (1) FR1-maintained cocaine (0.4 mg/kg) self-administration, (2) progressive ratio (PR)-maintained cocaine (0.2-1.6 mg/kg) self-administration, and (3) saline-induced and cocaine (10 mg/kg, IP)-induced reinstatement of drug seeking behavior were examined. RESULTS: HiS LgA rats escalated their cocaine intake more rapidly and self-administered more cocaine (mg/kg) than LoS LgA rats; however, there was no LoS versus HiS phenotype difference in the number of infusions self-administered by Day 21. Post-escalation cocaine self administration under an FR1 schedule did not differ as a function of phenotype (LoS versus HiS) or access condition (ShA versus LgA); however, LoS rats responded more for cocaine under the PR schedule than HiS rats, and they showed a greater reinstatement of cocaine-seeking behavior than HiS rats. In contrast, ShA versus LgA did not affect PR or reinstatement performance in the LoS and HiS groups. CONCLUSIONS: These results suggest that LoS and HiS rats have distinct drug-seeking and drug-taking profiles that differ as a function of the experimental phase and access condition. The LoS and HiS rats differ along a wide range of behavioral dimensions and represent an important model to study the interactions of feeding, emotionality, and other factors related to vulnerability to drug abuse. PMID- 15338103 TI - Neurocognitive effects of methylphenidate in adult attention deficit/hyperactivity disorder. AB - RATIONALE: Features of childhood attention-deficit/hyperactivity disorder (ADHD) often persist into adulthood. It has been shown that adult ADHD is associated with various neurocognitive deficits, including impairments in spatial working memory (SWM) and attention. It is not known whether these deficits are ameliorated by methylphenidate in adult ADHD. OBJECTIVES: The aim of this study was to evaluate the neurocognitive effects of a single dose of methylphenidate on SWM, visual memory, spatial span and sustained attention in adult ADHD. METHODS: Twenty-four adult patients, recruited from a specialised clinic for the assessment of adult ADHD, were entered into a double-blind, randomised, placebo controlled crossover study using a single 30 mg dose of methylphenidate. RESULTS: Eighteen patients met DSM-IV criteria for adult ADHD. Methylphenidate resulted in an improvement in SWM performance and sustained attention, together with a speeding in response time, in these patients. Six patients with attentional difficulties, who did not meet a DSM-IV diagnosis of ADHD, showed a different pattern of response to methylphenidate compared to the ADHD group. For the combined group, moderate correlations were shown between childhood ratings of ADHD (both self-reported and informant ratings) and response to methylphenidate on the SWM task. CONCLUSIONS: Adults with ADHD had a similar neurocognitive response to methylphenidate to that previously reported for childhood ADHD. Our results provide further support for the validity of the ADHD syndrome as defined by DSM-IV and indicate possible neurocognitive substrates for clinical improvement with chronic methylphenidate. PMID- 15338104 TI - Impulsivity (delay discounting) as a predictor of acquisition of IV cocaine self administration in female rats. AB - RATIONALE: Previous research in humans suggests a relationship between drug abuse and impulsivity as shown by selection of a smaller immediate reward over a larger delayed reward. However, it is not clear whether impulsivity precedes drug abuse or drug abuse influences impulsivity. OBJECTIVE: The hypothesis of the present experiment was that rats selected for choosing smaller, immediate over larger, delayed food would acquire IV cocaine self-administration faster than those choosing larger, delayed food rewards. METHODS: Female rats were screened for locomotor activity and trained on a delay discounting procedure that allowed them access to two response levers and a food pellet dispenser. Under a fixed-ratio (FR) 1 schedule, responding on one lever resulted in immediate delivery of one 45 mg pellet, while responding on the other lever resulted in delivery of three 45 mg pellets after a variable delay that increased after responses on the delay lever and decreased after responses on the immediate lever. For each rat, a mean adjusted delay (MAD) was calculated for each daily session, and stability was defined as MADs varying less than 5 s across 5 days. Based on their average MADs, rats were separated into low impulsive (LoI) and high impulsive (HiI) groups, implanted with an indwelling IV catheter, and trained to lever press for cocaine (0.2 mg/kg) under an FR1 schedule. RESULTS: There were no differences in locomotor activity between the LoI and HiI groups; however, a greater percentage of the HiI group acquired cocaine self-administration, and they did so at a significantly faster rate than the LoI rats. CONCLUSIONS: Performance on the delay discounting model of impulsivity predicted vulnerability to subsequent acquisition of cocaine self-administration. PMID- 15338105 TI - Ethanol consumption reduces the adverse consequences of self-administered intravenous cocaine in rats. AB - RATIONALE: Human drug users report that the initial positive effects of cocaine are followed by a dysphoric state characterized by anxiety and drug-craving. As a means of presumably attenuating these negative aftereffects, 50-90% of cocaine users choose to co-administer ethanol during cocaine binges. This co administration reportedly prolongs the "high" and diminishes the "low" associated with cocaine use. OBJECTIVE: The current study was intended to assess whether this phenomenon could be modeled in the animal laboratory. We have previously shown that animals running a straight alley for an intravenous cocaine reward develop a unique approach-avoidance "conflict" behavior that is characterized by stop and retreat behaviors as the subjects approach the goal box. The retreats are thought to reflect the concurrent positive (reward) and negative (anxiety) associations with the goal box and can be dose-dependently reduced by pretreatment with diazepam, which presumably attenuates the anxiety stemming from the conflict. METHODS: To test the role of ethanol in reducing cocaine-induced anxiety, rats were trained to run a straight-arm alley for a single daily injection of cocaine (1.0 mg/kg IV). RESULTS: Rats that had the opportunity to then drink either an 8% or a 4% sucrose-ethanol solution immediately following their daily runway trial came to exhibit fewer retreats than rats that did not drink ethanol following their cocaine injection. CONCLUSIONS: These results suggest that ethanol effectively reduces the development of approach-avoidance conflict in animals running an alley for IV cocaine, a result that may account for the prevalence of cocaine-ethanol co-administration in humans. PMID- 15338106 TI - In the ventral tegmental area picrotoxin blocks FGIN 1-27-induced increases in sexual behavior of rats and hamsters. AB - RATIONALE AND OBJECTIVES: There are two types of benzodiazepine receptors, mitochondrial benzodiazepine receptors (MBRs), and gamma-aminobutyric acid (GABA(A))/benzodiazepine receptor complexes (GBRs). MBR activation increases neurosteroidogenesis. Ventral tegmental area (VTA) infusions of the MBR agonist, FGIN 1-27, increase midbrain levels of the progesterone metabolite 5alpha-pregnan 3alpha-ol-20-one (3alpha,5alpha-THP) and lordosis of rats and hamsters. Activation of GBRs leads to membrane hyperpolarization. In the VTA, infusions of GBR agonists enhance 3alpha,5alpha-THP-facilitated lordosis. Thus, if, in the VTA, MBR-mediated increases in 3alpha,5alpha-THP enhance sexual responses via actions at GBRs, then blocking GBRs with picrotoxin will reduce FGIN 1-27-induced increases in sexual behavior of female rodents. METHODS: Ovariectomized rats and hamsters, with unilateral guide cannula to the VTA, received estradiol benzoate (10 mug; EB) at hour 0. Hamsters also received progesterone (100 mug) at hour 44. At hour 47.5, all animals were infused first with 10 ng or 20 ng picrotoxin or saline, vehicle to the VTA and, 30 min later, with 5 mug/11.4 nM FGIN 1-27 or saline, vehicle. Ten minutes later, animals were tested for sex and motor behavior. RESULTS: Picrotoxin, but not vehicle, infusions blocked FGIN 1-27 mediated increases in lordosis of rats and hamsters, proceptivity of rats, and sexual responsiveness of hamsters. In addition, midbrain 3alpha,5alpha-THP levels were higher in animals that received VTA infusions of FGIN 1-27, compared to those infused with saline, vehicle. CONCLUSION: In the VTA, GBRs are required for MBR-enhanced sexual behavior of EB-primed rats and EB- and progesterone-primed hamsters. PMID- 15338107 TI - Deficits in a sustained attention task following nicotine withdrawal in rats. AB - OBJECTIVE: Behavioural consequences of spontaneous and antagonist-precipitated withdrawal from nicotine upon performance of rats were compared alongside non nicotinic antagonists in the 5-choice serial reaction time task (5-CSRTT). METHODS: Male hooded Lister rats were trained to detect and respond to brief flashes of light presented every 15 s in one of five holes until a stable level of performance was achieved. RESULTS: Surgical removal of osmotic minipumps from rats having received nicotine (3.16 mg/kg per day base SC) chronically for 7 days produced marked deficits in performance. Compared to saline-treated controls, deficits were apparent 10 h and 16 h following nicotine abstinence; the percentage of omission errors increased concomitantly with modest decreases in response accuracy. Tests conducted 34 h and 106 h post-withdrawal indicated a progressive and complete recovery in attention performance, respectively. In another experiment, following the exposure to the same nicotine regime, administration of the competitive nicotine receptor antagonist dihydro-beta erythroidine precipitated immediate deficits in performance that were greater than those observed in saline-treated subjects. Methyllycaconitine, an alpha(7) nicotinic receptor antagonist failed to precipitate attention deficits in nicotine-treated rats. Tests with SCH23390 and raclopride produced impairments that were similar in profile to nicotine withdrawal contrasting with non-specific effects of dizocilpine. CONCLUSIONS: These results provide evidence of a cognitive impairment resulting from nicotine deprivation in rodents. Specifically, blockade of D(1) receptors by SCH23390 produced decrements in performance that were qualitatively similar but greater in magnitude to the alterations observed following nicotine withdrawal. Overall, assessing nicotine withdrawal in the 5-CSRTT presents an animal model that exhibits robust construct and face validity. PMID- 15338108 TI - Effects of norfluoxetine on the action potential and transmembrane ion currents in canine ventricular cardiomyocytes. AB - Norfluoxetine is the most important active metabolite of the widely used antidepressant compound fluoxetine. Although the cellular electrophysiological actions of fluoxetine are well characterized in cardiac cells, little is known about the effects of its metabolite. In this study, therefore, the effects of norfluoxetine on action potential (AP) configuration and transmembrane ion currents were studied in isolated canine cardiomyocytes using the whole cell configuration of patch clamp techniques. Micromolar concentrations of norfluoxetine (1-10 microM) modified AP configuration: amplitude and duration of the AP and maximum velocity of depolarization were decreased in addition to depression of the plateau and elimination of the incisura of AP. Voltage clamp experiments revealed a concentration-dependent suppression of both L-type Ca(2+) current, I(Ca) (EC(50)=1.13+/-0.08 microM) and transient outward K(+) current, I(to) (EC(50)=1.19+/-0.17 microM) having Hill coefficients close to unity. The midpoint potential of the steady-state inactivation of I(Ca) was shifted from 20.9+/-0.75 mV to -27.7+/-1.35 mV by 3 microM norfluoxetine ( P<0.05, n=7). No such shift in the steady-state inactivation curve was observed in the case of I(to). Similarly, norfluoxetine caused no change in the steady-state current voltage relationship of the membrane or in the density of the inward rectifier K(+) current, I(K1). All these effects of norfluoxetine developed rapidly and were fully reversible. Comparing present results with those obtained previously with fluoxetine, it can be concluded that norfluoxetine displays stronger suppression of cardiac ion channels than fluoxetine. Consequently, the majority of the cardiac side effects observed during fluoxetine treatment are likely to be attributed to its metabolite norfluoxetine. PMID- 15338109 TI - The small GTPase Rac is involved in clustering of hippocampal neurons and fasciculation of their neurites. AB - In hippocampal neurons cultured from brains of newborn rats, the glutamate receptor agonist N-methyl-D-aspartate induced the clustering of neuronal perikarya and the fasciculation of neurites. In addition, N-methyl-D-aspartate activated the small GTPase Rac1. Other stimuli of Rac activity, such as the Rho kinase inhibitors Y-27632, H-1152, and H89, as well as the cytotoxic necrotizing factor-1 from Escherichia coli, also caused neuronal clustering and neurite bundling. In neurons transiently transfected with dominant negative Rac1N17 neither N-methyl-D-aspartate nor Y-27632 induced clustering and fasciculation. In addition, the PI3-kinase inhibitors wortmannin and LY-294002 prevented these effects, as did a dominant negative form of p110PI3-Kgamma. Time-lapse microscopy showed that lethal toxin from Clostridium sordellii, which inhibits Rac, and wortmannin blocked the neuronal migration induced by Y-27632. In contrast, only lethal toxin reversed the clustering and fasciculation induced by pre-treatment with Y-27632. This effect of the toxin may be due to inactivation of Ras, since FTI-277, which prevents the farnesylation of Ras and thereby inactivates the GTPase, also dissolved the preformed clusters. We suggest that active Rac and a PI3-kinase synergistically induce neuronal migration, whereas a Ras isoform is responsible for the lasting attachment of neurons necessary for clustering and neurite fasciculation. PMID- 15338110 TI - 3,4,5,6-Tetrahydroxyxanthone prevents vascular endothelial cell apoptosis induced by high glucose. AB - Apoptosis of endothelial cells may be an important risk factor contributing to the incidence of vascular complications in diabetes. In the present study, we tested the effect of 3,4,5,6-tetrahydroxyxanthone, a synthetic xanthone derivative, on apoptosis induced in human umbilical vein endothelial cells (HUVEC) by a high glucose concentration. Cell apoptosis was detected using DNA ladder formation and flow cytometric techniques. The expression of Bcl-2 protein was analysed using flow cytometric techniques. Lactate dehydrogenase (LDH) activity and malonyldialdehyde (MDA) content in the medium were measured. Cell viability was assayed by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) method. Exposure of HUVEC to a high glucose concentration (30 mM) for 48 h markedly increased LDH release and MDA content in the medium and induced apoptosis and Bcl-2 protein expression in HUVEC. Pretreatment with 3,4,5,6 tetrahydroxyxanthone (1, 3 or 10 microM) or probucol (10 microM) significantly decreased the level of LDH and MDA in the medium, reduced apoptosis and increased the expression of Bcl-2 protein in HUVEC. These results suggest that 3,4,5,6 tetrahydroxyxanthone inhibits high-glucose-induced endothelial cell apoptosis by increasing Bcl-2 protein expression in HUVEC. PMID- 15338111 TI - Chemolithoautotrophy in the marine, magnetotactic bacterial strains MV-1 and MV 2. AB - Magnetite-producing magnetotactic bacteria collected from the oxic-anoxic transition zone of chemically stratified marine environments characterized by O2/H2S inverse double gradients, contained internal S-rich inclusions resembling elemental S globules, suggesting they oxidize reduced S compounds that could support autotrophy. Two strains of marine magnetotactic bacteria, MV-1 and MV-2, isolated from such sites grew in O2-gradient media with H2S or thiosulfate (S2O3(2-)) as electron sources and O2 as electron acceptor or anaerobically with S2O3(2-) and N2O as electron acceptor, with bicarbonate (HCO3-)/CO2 as sole C source. Cells grown with H2S contained S-rich inclusions. Cells oxidized S2O3(2-) to sulfate (SO4(2-)). Both strains grew microaerobically with formate. Neither grew microaerobically with tetrathionate (S4O6(2-)), methanol, or Fe2+ as FeS, or siderite (FeCO3). Growth with S2O3(2-) and radiolabeled 14C-HCO3- showed that cell C was derived from HCO3-/CO2. Cell-free extracts showed ribulose 1,5 bisphosphate carboxylase/oxygenase (RubisCO) activity. Southern blot analyses indicated the presence of a form II RubisCO (cbbM) but no form I (cbbL) in both strains. cbbM and cbbQ, a putative post-translational activator of RubisCO, were identified in MV-1. MV-1 and MV-2 are thus chemolithoautotrophs that use the Calvin-Benson-Bassham pathway. cbbM was also identified in Magnetospirillum magnetotacticum. Thus, magnetotactic bacteria at the oxic-anoxic transition zone of chemically stratified aquatic environments are important in C cycling and primary productivity. PMID- 15338112 TI - Trabecular bone is more deteriorated in spinal cord injured versus estrogen-free postmenopausal women. AB - The prevalence of osteoporosis is high among postmenopausal women and individuals sustaining a spinal cord injury (SCI). We assessed the effects of estrogen loss and unloading on the trabecular bone of the knee in women. Pre- and postmenopausal ambulatory women (n=17) were compared to pre- and postmenopausal women with SCI (n=20). High-resolution magnetic resonance imaging was used to compare groups on apparent measures of trabecular bone volume, trabecular number, trabecular spacing, and trabecular thickness in the distal femur and proximal tibia, regions with a high proportion of trabecular bone and the most common fracture site for SCI patients. Trabecular bone was deteriorated in women with SCI compared to ambulatory women. SCI groups had fewer, (-19 and -26% less) and thinner trabeculae (-6%) that were spaced further apart (40% and 62% more space between structures) resulting in less trabecular bone volume (-22% and -33%) compared to the ambulatory groups (tibia and femur, respectively). Postmenopausal women with SCI also had 34% greater trabecular spacing in the tibia compared to the 40-year-old premenopausal women with SCI, showing an interaction between unloading and estrogen loss. Middle-aged postmenopausal, ambulatory women, not taking estrogen or medications that affect bone, did not show the deteriorated trabeculae that were evident in women with SCI, nor did they show differences in distal femur and proximal tibia trabeculae compared to a premenopausal group. We conclude that the effect of unloading on bone architecture is greater than that of estrogen loss in middle-aged women. PMID- 15338113 TI - Bone outcomes and technical measurement issues of bone health among children and adolescents: considerations for nutrition and physical activity intervention trials. AB - Weight-bearing physical activity and calcium intake are two important behavioral influences for bone health. Physical-activity and calcium-intake intervention trials with youth have been implemented to evaluate their efficacy and effectiveness to decrease the risk for subsequent osteoporosis and fractures. Technical aspects of bone measurement have not been routinely reported in published trials of youth, even though they can have an impact on study findings and interpretation. This paper provides an overview of the outcome variables reported, and technical issues--such as software and bone detection, growth, and movement--that affect bone mass measurements among children and adolescents. It describes the implications of these issues for the interpretation of intervention effects observed in intervention trials, and provides recommendations for future research. PMID- 15338114 TI - Effects of tamoxifen on Doppler velocimetry parameters of periurethral vessels in postmenopausal women. AB - The purpose of the present study was to assess the effect of tamoxifen on periurethral vessels by Doppler velocimetry examination. Increase in the number of these vessels as well as decrease in resistance and pulsatility indices by tamoxifen were observed. PMID- 15338115 TI - Transplant quality in Italy: analysis of the 1995-2000 period. AB - Evaluation of outcomes is a major step in quality assessment of any health process. In the transplant field, the evaluation of outcome is extremely important for both patients' growing demand for health and for the joint commitment the transplant process requires. In this study, the outcome of 12,647 transplants, carried out between 1995 and 2000 were analysed. Graft survival at 5 years was 79% for kidney, 67% for liver, 72% for heart and 38% for lung. Patient survival was 92% for kidney, 76% for liver, 72% for heart and 38% for lung. In comparison to other international case records [Collaborative Transplant Study (CTS) and The United Network for Organ Sharing (UNOS)], results are similar or even better for all transplant programmes. As a whole, survival after solid organ transplant in Italy ranks among the best for both donations and transplantation. The quality of transplants carried out is above European standards. Nevertheless, the growing health needs of patients require improvement in both the procurement process and in the use of available organs. PMID- 15338116 TI - Prevalence and correlates of depression symptoms at 10 years after heart transplantation: continuous attention required. AB - This study investigates the presence and correlates of symptoms of depression at 5 and 10 years after heart transplantation, with particular attention given to patients showing symptoms at both time points. Prevalence of depression symptoms were studied in 41 patients, prospectively, at 5 and 10 years after heart transplantation. We examined potential correlates of depression symptoms (i.e. worse functional capacity, inadequate coping mechanisms and lack of social contacts) 10 years after transplantation. The prevalence of depression symptoms was 30% at 5 years and 22% at 10 years. Of the 41 patients, 20% were depressed at both time points. Those patients had significantly higher scores on passive coping and had significantly lower club membership. They also tended to have more negative emotions (i.e. anger, hostility and irritability) and less engagement in sports activities. Functional capacity was not different. Depression symptoms were prevalent and persistent in the long-term after heart transplantation. This study opens perspectives for beyond-standard pharmacological and psychotherapeutic treatment for depression, i.e. training patients who are using passive coping to use problem-solving capacities instead and motivating them to engage in social life and sports activities. PMID- 15338117 TI - The relative importance of cyclosporine exposure in heart, kidney or liver transplant recipients on maintenance therapy. AB - We investigated the relationship between cyclosporine exposure and the presence of cyclosporine-related side effects and assessed the advantage of the cyclosporine concentration 2 h post-dose (C(2)) over pre-dose concentration (C(0)) monitoring. Cyclosporine area-under-the-concentration-time curves were measured during the absorption phase (AUC(0-4 h)) in 49 liver, 28 heart and 26 kidney transplant recipients (time since transplantation >6 years) with or without cyclosporine-related side effects on maintenance therapy. The cyclosporine C(0) correlated well with AUC(0-4) (r=0.77), whereas C(2) levels correlated strongly with AUC(0-4) (r=0.92). Although we observed a trend towards higher CsA concentrations in transplant recipients with side effects than in patients without CsA toxicity, the large majority of those differences were not statistically significant. Thus, as cyclosporine exposure was not clearly related to the presence of side effects, and C(0) correlated fairly with AUC(0-4), the advantage of monitoring cyclosporine treatment using C(2) rather than C(0), may be limited for patients on cyclosporine maintenance therapy. PMID- 15338118 TI - Liver transplantation using organs from donors older than 60 years. AB - At present, it is frequently accepted to expand the organ pool for liver transplantation (LTx) by including livers from critical donors. From 1990 to June 2002 a total of 1,208 LTx were performed. Of those, 67 livers from donors older than 60 years were transplanted to 66 patients, including re-LTx in eight patients. Fourteen patients had malignant diseases (21%). Ten patients had a high urgency status (15%). Median donor age was 65 years (range 61-80 years). Primary graft function was observed in 84%. Patient survival rate at 1 and 5 years was 79% and 62%, and graft survival was 68% and 53%, respectively. No difference was observed in LTx with livers from donors younger than 60 years. Fifteen graft losses occurred during the study. Surgical complications were observed in 23 patients (34%). The outcome of LTx with livers from donors older than 60 years is satisfactory and is comparable to results of LTx with livers from donors younger than 60 years. The frequency of vascular complications and cholestasis syndrome is not increased. PMID- 15338119 TI - Polyclonal anti-thymocyte globulins influence apoptosis in reperfused tissues after ischaemia in a non-human primate model. AB - Reperfusion triggers the expression of inflammatory cytokines and adhesion molecules that increase the rate of apoptosis in the reperfused tissues after ischaemia, thus worsening the outcome of the grafts. Polyclonal anti-thymocyte globulins (pATGs) are able to reduce the number of lymphocytes as well as block adhesion molecules and induce apoptosis in T-lymphocytes through Fas-ligand. The aims of this study were to investigate the influence of pATGs on the prevention of apoptosis of reperfused tissues after ischaemia and to monitor their capability to enhance lymphocyte apoptosis thus decreasing the deleterious effects of ischaemia/reperfusion injury (IRI). Extremities of cynomolgus monkeys ( n=8) were flushed via either the femoral or the brachial artery. After 60 min of ischaemia the limbs were reperfused with human blood. ATG was added to the blood in a therapeutic dose 20 min prior to reperfusion of the extremities. Surgically available limbs ( n=20) were assigned to the following groups: ATG group ( n=10) and control group (without ATG; n=10). DNA fragmentation analysis was performed in situ to detect apoptosis at the single-cell level. Our study shows an increased rate of muscle and connective tissue apoptosis in the control group compared with the ATG-treated group. Cells found in the vascular areas present different rates of apoptosis, with enhanced cellular death of endothelium and connective perivascular areas being observed in the control group. The group treated with ATG shows an increased rate of white blood cell (WBC) apoptosis in vascular and perivascular areas. Previous studies have shown that pATGs are able to induce apoptosis as well as complement-mediated cell death in peripheral T lymphocytes in vitro. Our results confirm that pATGs not only increase the rate of apoptosis of WBCs in vivo but also have a protective effect on the reperfused tissue. This may alleviate the damage after reperfusion of solid-organ transplantation. PMID- 15338120 TI - Improvement in long-term graft survival in cadaveric renal transplant recipients treated with mycophenolate mofetil. AB - Though mycophenolate mofetil has markedly reduced the incidence of acute rejection in renal transplantation, a significant improvement in graft survival has been more difficult to demonstrate. This retrospective study compares an historical control group of 210 consecutive renal transplant patients, who had received ATG induction associated with cyclosporin, prednisolone and azathioprine, with 187 patients receiving mycophenolate instead of azathioprine. The incidence of acute rejection was decreased with mycophenolate. In rejection free patients, the 3-year graft survival rates were equivalent. In contrast, graft survival at 3 years improved significantly for patients who experienced a rejection crisis and remained under the initial triple drug regimen with mycophenolate compared to the patients of the historical group who were kept on azathioprine after a rejection episode. In conclusion, mycophenolate mofetil is not only able to reduce the incidence of acute rejection but could also improve the prognostic significance of acute rejection crises. PMID- 15338121 TI - Effect of sirolimus on renal ischaemia/reperfusion injury in normotensive and hypertensive rats. AB - BACKGROUND: Renal ischaemia/reperfusion (I/R) injury and hypertension represent major alloantigen-independent factors contributing to the development of chronic allograft nephropathy of renal allografts. In the present study, we investigated the effect of the anti-proliferative immunosuppressant, sirolimus (SIR), in a model of accelerated renal injury in hypertensive transgenic rats (TGRs). Twenty anaesthetized uninephrectomized TGRs with renin overproduction [TGR(mREN2)27] and 20 normotensive Han SD (SD) rats as genetic controls had their renal pedicles clipped for 45 min and were subsequently treated with either SIR (0.5 mg/kg per day, orally) or placebo ( n=10 in each group) for 16 weeks, after which time the kidneys were harvested for morphological and immunohistochemical analysis. High renin hypertension aggravated the functional and structural changes induced by I/R in SD animals: both SIR-treated and untreated TGRs exhibited significantly greater proteinuria and suffered from more severe glomerulosclerosis ( P<0.01) and vasculopathy ( P<0.01), as well as compensatory renal hypertrophy ( P<0.01) and tissue TGF-beta1 expression, than both normotensive SD groups ( P<0.01). SIR treated SD rats showed reduced proteinuria ( P<0.01), glomerulosclerosis ( P<0.01), and TGF-beta1 expression in the glomerular epithelium and proximal tubuli ( P<0.05) compared with placebo-treated SD rats. SIR-treated TGRs had significantly lower proteinuria at week 4 after I/R ( P<0.01) than placebo treated TGRs, but there were no significant differences thereafter. Morphological patterns were similar in treated and untreated TGRs at week 16. High-renin induced hypertension aggravated the renal injury induced by I/R. Sirolimus treatment ameliorated some late functional and morphological changes induced by I/R injury in hypertensive TGRs but, particularly, in normotensive SD rats. PMID- 15338122 TI - Why do some diabetic patients on the kidney transplant waiting list not receive a transplant? AB - The waiting list (WL) history of 405 diabetic patients placed on the kidney transplantation WL for the years 1993-2000 was examined. By 31 December 2000, 295 (73 %) patients had received a transplant. Of the remaining 110 patients 53 (13 %) were still on the WL; 27 of these were temporarily withdrawn, i.e. non-active, 46 others (11 %) had died and 11 (3 %) had been permanently removed. Patient follow-up continued until the end of 2002. Although the mean total time on the WL of the non-transplanted was twice that of the transplanted patients there were no significant differences in the mean active times on the WL. The mean cumulative withdrawal time of the transplanted and those on the active WL was less than 10 % of their total time on the list, but for the patients who had died or were withdrawn on 31 December 2000 it exceeded 50 %, usually because of diabetic complications. The 5-year survival of the transplanted patients was greatly superior to that of the non-transplanted, as expected. However, the better survival of the transplanted patients is not necessarily proof of a better treatment modality but rather a consequence of the exclusion from transplantation of patients suffering from diabetic complications. It is not justified to compare the survival of transplantable and non-transplantable WL patients. PMID- 15338123 TI - Brain death. PMID- 15338124 TI - Dexmedetomidine infusion for more than 24 hours in critically ill patients: sedative and cardiovascular effects. AB - OBJECTIVE: To assess the potential of dexmedetomidine for targeted sedation in complex Intensive Care (ICU) patients for >24 h. DESIGN: Prospective, open label, clinical trial. SETTING: Tertiary general ICU. PATIENTS: Twenty critically ill patients, mean APACHE II 23(+/-9). INTERVENTIONS: A continuous infusion of dexmedetomidine, median infusion time 71.5 (35-168) h, starting at 0.4 microg.kg.h without a loading dose and adjusted (0.2-0.7 microg.kg.h) to a target Ramsay Sedation Score (RSS) of 2-4. Rescue midazolam and/or morphine/fentanyl were given as clinically indicated. MEASUREMENTS AND RESULTS: Haemodynamic parameters and RSSs were collected until 24 h after cessation. An RSS 2-5 was achieved in 1,147 (83%) of observations with a reduction in RSS of 6 from 13% in the first 6 h to 3% between 18 h and 24 h. Sixteen patients needed minimal or no additional midazolam, median 4 mg/day (0.5-10) and ten required minimal or no additional analgesia, median 2 mg/day (0.5-4.5), 55 microg/day (14-63) of morphine/fentanyl. RESULTS: A 16% reduction in mean systolic blood pressure (SBP) and 21% reduction in heart rate (HR) occurred over the first 4 h followed by minimal (+/- 10%) changes throughout the infusion. A rise in SBP was observed in two patients. After abrupt cessation, SBP and HR monitored for 24 h rose by 7% and 11%, respectively. CONCLUSIONS: Dexmedetomidine was an effective sedative and analgesic sparing drug in critically ill patients when used without a loading dose for longer than 24 h with predictable falls in blood pressure and HR. There was no evidence of cardiovascular rebound 24 h after abrupt cessation of infusion. PMID- 15338125 TI - Effects of thoracic epidural anaesthesia on intestinal microvascular perfusion in a rodent model of normotensive endotoxaemia. AB - OBJECTIVE: To investigate whether sympathetic blockade by means of thoracic epidural anaesthesia (TEA) increases intestinal perfusion during normotensive endotoxaemia. DESIGN: A prospective, randomised and controlled animal study. SETTING: Animal laboratory in a university hospital. SUBJECTS: Sprague-Dawley male rats. INTERVENTIONS: The rats were anaesthetised with urethane and ketamine, mechanically ventilated and haemodynamically monitored. Lidocaine or saline were infused continuously via thoracic epidural catheters followed by a continuous intravenous infusion of Escherichia coli lipopolysaccharide (1.5 mg/kg per h). Densities of perfused and non-perfused capillaries (i.e., with and without erythrocyte perfusion, respectively) as well as erythrocyte velocity in both the mucosa and the muscularis of the terminal ileum were determined using intravital microscopy. MEASUREMENTS AND RESULTS: Measurements were performed at baseline, after 30 min of epidural infusion as well as after 60 and 120 min of lipopolysaccharide infusion. In animals receiving TEA, mean arterial pressure and heart rate were significantly reduced throughout the experiment. In the muscularis the endotoxaemia-induced increase in non-perfused capillaries was absent with epidural lidocaine (0 [0/0] versus 39 [36/137] cm(-1), median [25(th)/75(th) percentile]), whereas in the mucosa perfused capillary density declined to a greater extent than in controls (-47 [-53/-23]%) versus -19 [ 34/+10]%, p<0.05). Erythrocyte velocity decreased with endotoxaemia and was not influenced by epidural lidocaine. CONCLUSIONS: Microvascular perfusion data during endotoxaemia show a redistribution of blood flow towards the mucosa. TEA seems to impede this redistribution resulting in improved muscularis and worsened mucosal microvascular perfusion. PMID- 15338127 TI - Glomerular structural and functional changes in a high-fat diet mouse model of early-stage Type 2 diabetes. AB - AIMS/HYPOTHESIS: Type 2 diabetes often results in diabetic nephropathy, which is preceded by an elevated glomerular filtration rate (GFR). This study was designed to develop a mouse model of Type 2 diabetes and to elucidate the glomerular events in the early stages of diabetic nephropathy. METHODS: Four-week-old mice were fed a normal or high-fat (42% of total calories from fat) diet, and body weight, blood glucose, insulin, leptin, lipids and GFR were monitored from 9 to 21 weeks or longer after the feeding programme. Mesangial cell dedifferentiation was accessed by alpha-smooth muscle actin staining. Glomerular hypertrophy was determined using image analysis with haematoxylin-eosin staining. Matrix deposition was determined by type IV collagen staining. RESULTS: After 9 weeks, mice fed a high-fat diet weighed more than mice fed a normal diet (30.5+/-1.2 vs 22.3+/-0.5 g, p<0.05), and mice fed a high-fat diet were hyperinsulinaemic (283.9+/-69.7 vs 102.9+/-36.4 pmol/l, p<0.05), hyperglycaemic (8.0+/-0.6 vs 6.5+/ 0.2 mmol/l, p<0.05) and their leptin levels were increased six-fold (1.48+/-0.45 vs 0.25+/-0.03 ng/ml, p<0.05). After 13 weeks, mice fed a high-fat diet showed hyperfiltration (GFR; 440+/-60 vs 210+/-10 microl/min, p<0.05). During the early stages of diabetic nephropathy, mesangial cell dedifferentiation was evident, shown by increased expression of alpha-smooth muscle actin in the glomeruli. After 9 weeks, mice fed a high-fat diet already demonstrated increased type IV collagen deposition. After 13 weeks, they developed enlarged glomerular tufts compared with those of their age-matched controls. CONCLUSIONS/INTERPRETATION: The results of this study suggest that collagen IV deposition precedes the hyperfiltration and enlargement of glomeruli in early-stage diabetic nephropathy. Dedifferentiation of mesangial cells may be associated with collagen IV deposition. PMID- 15338128 TI - Albuminuria is a marker of increasing intracranial and extracranial vascular involvement in Type 2 diabetic Chinese patients. AB - AIMS/HYPOTHESIS: Albuminuria has been reported to be a marker of cardiovascular risk factors and disease morbidity and mortality, but its relationship with intracerebral atherosclerotic disease is less clear. The aim of this study was to investigate the association between albuminuria and intracranial and extracranial vascular involvement in Chinese Type 2 diabetic patients. METHODS: The anthropometric and fasting biochemical measurements of 966 Type 2 diabetic patients with normoalbuminuria (55.6%), microalbuminuria (27.7%) or macroalbuminuria (16.7%) were compared. The prevalence of microvascular and macrovascular disease and middle cerebral artery (MCA) stenosis, measured by transcranial Doppler ultrasound, were also compared between the groups. RESULTS: Albuminuria was closely associated with a range of adverse parameters, including high BP, dyslipidaemia, smoking and adiposity (all p<0.01). The prevalence of microvascular disease (retinopathy p<0.001) and macrovascular disease (peripheral vascular disease p=0.012, myocardial infarction, p=0.004, MCA stenosis p<0.001) increased significantly with increasing levels of albuminuria. Albuminuria was also found to be an independent predictor of microvascular and macrovascular disease. CONCLUSIONS/INTERPRETATION: Albuminuria was an independent predictor of increasing levels of vascular risk factors and microvascular and macrovascular disease in this group of Type 2 diabetic patients, and a possible role for albuminuria as a marker of intracranial cerebrovascular disease should be further investigated. PMID- 15338129 TI - Asymptomatic cytomegalovirus infection is associated with increased risk of new onset diabetes mellitus and impaired insulin release after renal transplantation. AB - AIMS/HYPOTHESIS: The human cytomegalovirus (CMV) may increase the risk of diabetes mellitus, but the literature is scarce. The present study was designed to test the hypothesis that asymptomatic CMV infection is associated with increased risk of new-onset diabetes after renal transplantation, and to assess the impact of asymptomatic CMV infection on OGTT-derived estimates of insulin release and insulin action. METHODS: A total of 160 consecutive non-diabetic renal transplant recipients on cyclosporine (Sandimmun Neoral)-based immunosuppression were closely monitored for CMV infection during the first 3 months after transplantation. All patients underwent a 75-g OGTT at 10 weeks. Excluded from the analyses were 36 patients with symptomatic CMV infection (disease). RESULTS: The incidence of new-onset diabetes was 6% in a control group of recipients without CMV infection (4/63) and 26% in the group with asymptomatic CMV infection (16/61). Asymptomatic CMV infection was associated with a significantly increased risk of new-onset diabetes (adjusted odds ratio: 4.00; 95% CI: 1.19 to 13.43, p=0.025). The group of patients with CMV infection had a significantly lower median insulin release than controls. CONCLUSIONS/INTERPRETATION: Our findings support the hypothesis that asymptomatic CMV infection is associated with increased risk of new-onset post-transplant diabetes mellitus, and suggest that impaired insulin release may involve one pathogenetic mechanism. PMID- 15338130 TI - CMV prophylaxis: a useful step towards prevention of post-transplant diabetes? PMID- 15338131 TI - Self-incompatibility genotypes in almond re-evaluated by PCR, stylar ribonucleases, sequencing analysis and controlled pollinations. AB - As part of the almond breeding programme at IRTA, we investigated the S genotypes of several cultivars using a combination of RNase zymograms, testcrosses, pollen tube growth analysis and molecular identification by PCR analysis. For some of the cultivars examined, discrepancies appeared between their S alleles as reported in the literature and those found in this investigation, leading to a re evaluation of their S genotypes. Analysis of the stylar ribonucleases (RNases), which are known to correlate with S alleles, of cvs. Achaak, Ardechoise, Desmayo Largueta, Ferrastar, Gabaix, Garbi, Glorieta, Languedoc, Primorskiy and Texas revealed inconsistencies with respect to the S5 and S10 alleles. However, PCR with the conserved primer pair AS1II/AmyC5R failed to detect any of these inconsistencies. When the S alleles from Desmayo Largueta, Gabaix, Primorskiy and Texas were sequenced, Texas and Primorskiy were found to carry the reported S5 allele, while Desmayo Largueta and Gabaix carried a new allele, which has been tentatively denoted as S25 This new S allele, previously reported to be S10, was also identified in Achaak, Ardechoise and Ferrastar. The proposed new S genotypes are Achaak (S2S25), Ardechoise (S1S25), Desmayo Largueta (S1S25), Ferrastar (S2S25) and Gabaix (S10S25). The S alleles of Garbi, Glorieta, Languedoc, Texas and Primorskiy remain as reported in the literature. Testcrosses in the field and laboratory confirmed the new S genotypes. One cultivar (Gabaix) could be assigned to the existing cross-incompatibility group O of unique genotypes, and two new groups were established (XVI and XVII) consisting of two cultivars each. The clarification of these S alleles will be useful in almond breeding programmes and for planning new commercial orchards in the future. PMID- 15338132 TI - A cytogenetic method for stacking gene pairs in common wheat. AB - The potential for non-reciprocal Robertsonian translocations of wheat (Triticum aestivum L.) to assist in the stacking of genes was assessed from a study of their cytological and genetic behaviour. To obtain translocations, a double monosomic (3B+5A; 2n=40=19ii+2i) was crossed reciprocally with a contrasting disomic. Individuals inheriting a broken monosome were identified from the loss of one arm-specific DNA marker coupled with retention of a marker for the opposite arm. No double breaks (potential translocations) were found in 180 cross progeny recovered from pollen of the double monosomic but two instances (loss of 5AL plus 3BS; loss of 5AL plus 3BL) were found in 251 progeny recovered from ovules. Meiotic pairing and multi-color genome-specific fluorescence in situ hybridization (mcGISH) showed that each plant with a double break contained one translocated chromosome between the A and B genomes that had rejoined at the centromere and that formed a trivalent (19ii+ liii) in about 83% of PMC. Most trivalents (approximately 92%) aligned at metaphase in a 'V' configuration(alternate disjunction) while the rest aligned in linear 'I'(adjacent disjunction) or ambiguous 'L' configurations. Genetic analysis of a testcross of these 'fusion monosomics' showed that this preferential co orientation of the trivalent influenced the assortment of the chromosome arms involved. Loci that were located in the hemizygous ends of the 'V' trivalent showed strong quasi-linkage in that most ovules from the female testcross carried relevant DNA markers either from both standard chromosomes or from neither. This shows that, in most cases, the two standard chromosomes assorted to the same pole while the fused monosome segregated to the opposite pole. For heterozygous loci (present both on the fusion monosome and the standard chromosomes) assortment was either independent or showed partial linkage to the hemizygous arm depending on the reported recombination distance from centromere. Marker assortment was further distorted in male testcrosses and in doubled haploids (made from the fusion monosomics by the maize method) by the strong selective advantage of pollen or haploids that inherited the standard chromosomes rather than the deficiencies. This genetic data shows that under the combined influence of alternate disjunction and natural selection, progeny of fusion monosomics will revert to the standard disomic arrangement, fixing the gene content of both hemizygous arms in the process. Thus, any pair of genes could be targeted for joint fixation by isolating the fusion monosome that will link them temporarily in a segregating population. PMID- 15338133 TI - A novel major gene on chromosome 6H for resistance of barley against the barley yellow dwarf virus. AB - In a mapping population derived from the Ethiopian barley line L94 x Vada, natural infection by barley yellow dwarf virus (BYDV) occurred. While line L94 hardly showed symptoms, Vada was severely affected. The 103 recombinant inbred lines segregated bimodally. The major gene responsible for this resistance mapped to chromosome 6H. We propose to name the locus Ryd3. A subset of recombinant inbred lines, L94, and Vada were planted in a subsequent field test which confirmed the previous field observations. Double antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) indicated that the epidemic was due to a combination of the serotypes BYDV-PAV and BYDV-MAV. In the accessions with the least BYDV symptoms no virus was detected, justifying the consideration of the gene as conferring true resistance rather than tolerance to these viruses. In a laboratory/gauze house trial a near-isogenic line carrying the Vada chromosome 6H fragment in an L94 background was affected as much as Vada. The effect of Ryd3 was quantified, and compared with that of the only other known major gene for resistance to BYDV, Ryd2, which is also of Ethiopian origin and is located on chromosome 3H. Both genes seemed to reduce the chance of the viral isolate used in this study to establish infection. In plants in which it became established, the virus concentration reached a similar level as in susceptible accessions, but with less dramatic symptom development. Inoculated plants in which the virus failed to multiply tended to show an increase in the number of ears per plant, resulting in higher grain yield per plant. Ryd3 co-segregates with several PCR based molecular markers that may serve for marker assisted selection. PMID- 15338134 TI - Quantitative trait loci controlling phenotypes related to the perennial versus annual habit in wild relatives of maize. AB - We used quantitative trait locus/loci (QTL) mapping to study the inheritance of traits associated with perennialism in a cross between an annual (Zea mays ssp. parviglumis) and a perennial (Z. diploperennis) species of teosinte. The most striking difference between these species is that Z. diploperennis forms rhizomes, whereas Z. mays ssp. parviglumis lacks these over-wintering underground stems. An F2 population of 425 individuals was genotyped at 95 restriction fragment length polymorphism marker loci and the association between phenotype and genotype was analyzed by composite interval mapping. We detected a total of 38 QTL for eight traits. The number of QTL found for each trait ranged from two for rhizome formation to nine for tillering. QTL for six of the traits mapped near each other on chromosome 2, and QTL for four traits mapped near each other on chromosome 6, suggesting that these regions play an important role in the evolution of the perennial habit in teosinte. Most of the 38 QTL had small effects, and no single QTL showed a strikingly large effect. The map positions that we determined for rhizome formation and other traits in teosinte may help to locate corresponding QTL in pasture and turf grasses used as forage for cattle and for erosion control in agro-ecosystems. PMID- 15338135 TI - Identification of candidate genes for in vitro androgenesis induction in maize. AB - Extensive studies have been conducted to understand the genetic control of in vitro androgenesis, but little is know about the genes and the mechanisms involved in the switch that allows an immature pollen grain to develop as an embryo. We have developed two maize isogenic lines with high androgenetic aptitude, named AH5-44 and AH5-49, through backcross and selection from a high responsive DH229 line on the non-responding A188 line genetic background. The genomic structure of these two lines was precisely described with microsatellite markers. Five regions retained from the parent DH229 highly responsive to androgenesis were localised in both AH5-44 and AH5-49. Sequences expressed on microspores extracted from the four lines were amplified using a cDNA-AFLP protocol. For each line, eight culture conditions were compared: microspores extracted after tassel recovery, after 7 or 14 days in cold room and after 1-4 days of in vitro culture. This genetic and developmental screening allowed us to identify four sequences, including a new HSP70-like candidate gene. Possible implication of the identified sequences in androgenesis response is discussed. PMID- 15338136 TI - [Development of the Stettin municipal hospital. The beginnings of urology in Stettin (1.)]. AB - The growth of the city's population, rapid advances in medical science, the increasing understanding of the importance of hygiene and, finally, the demand for health care led to the erection of a modern 353-bed municipal hospital complex at Pommerensdorf, Apfelallee, which was officially opened in 1879. From the very beginning of its existence, the municipal hospital was constantly in the process of extension and rebuilding. A hospital base was set up and new departments and clinics were opened. In 1937, the number of the hospital beds had increased to 1,004. The official existence of an independent urology department with 30 beds, first headed by Dr. Felix Hagen (1880-1962), dates back to March 1919. The aim of this new department was to carry out examinations and differential diagnoses, together with the department of internal medicine, for those patients from the urological surgery sector including evaluation for surgery. Surgical urology was the responsibility of the surgery department. The remaining cases requiring urological experience, such as those involving transurethral intervention, diseases of the bladder and urinary tract infection, were treated in the new urology department. As an auxiliary department, de facto dependent on the departments of surgery and internal medicine, urology could not develop properly and survive. In October 1935, the position of the head of the urological department was terminated, and urology was incorporated into the department of surgery. During the Second World War, between 1939 and 1945, as during the World War 1, a field hospital was set up in the municipal hospital at Pommerensdorf in Stettin. PMID- 15338137 TI - [Diagnosis of lower urinary tract obstruction in man. Guidelines of the German Society for Urologists]. PMID- 15338138 TI - [Cardiovascular whole body MRI with parallel imaging]. AB - PURPOSE: To create a whole-body cardiovascular MRI protocol with parallel imaging (iPAT). MATERIAL AND METHODS: Twenty two persons participated in the whole body scan. Due to iPAT high resolution imaging of the heart could be performed. 3D contrast enhanced MR angiography (3D-CE-MRA) took only 62 s to cover the whole body at a spatial resolution of less than 1.4x1.0x1.5mm(3). Scan time for HASTE lung imaging could be reduced by the factor of 2 while maintaining the signal to noise ratio (SNR). Image quality was rated by 2 radiologists blinded to each other. RESULTS: Mean scan time was 104 min on the standard system and less than 80 min on the new system. 75% of all MRA segments were rated good in terms of vessel conspicuity, more than 80% had no venous overlay. One case of distal occlusion of the anterior tibial artery and two cases of myocardial infarction were detected. CONCLUSION: Parallel imaging offers the possibility of fast whole body imaging. A combination of morphologic and functional imaging can be performed within less than 80 min. PMID- 15338139 TI - [Autologous chondrocyte transplantation for the treatment of articular cartilage defects inf the knee joint. Techniques and results]. AB - BACKGROUND: Currently the use of autologous chondrocytes as a cartilage-repair procedure for the repair of injured articular cartilage of the knee joint, is recommended. METHODS: This review presents the technique of autologous chondrocyte transplantation (ACT) and their modifications as matrix-associated autologous chondrocyte transplantation (MACT). Beside the surgical procedure the experimental and clinical results are discussed. Furthermore the major complications and the indication guidelines are presented. RESULTS: Articular cartilage in adults has a poor ability to self-repair after a substantial injury. Surgical therapeutic efforts in treating cartilage defects have focused on bringing new cells capable of chondrogenesis into the lesions. With ACT good to excellent clinical results are seen in isolated posttraumatic lesions of the knee joint in the younger patient with the formation of hyaline-like repair tissue. The major complications are periosteal hypertrophy, delamination of the transplant, arthrofibrosis and transplant failure. The current limitations include osteoarthritic defects and higher patient age. CONCLUSION: With the right indication and operative technique ACT is an effective and save option for the treatment of large full thickness cartilage defect of the knee joint. PMID- 15338140 TI - [The so-called spontaneous low CSF pressure pressure syndrome. Case results indicating a disturbance in CSF/blood volume regulation]. AB - The new IHS classification describes under the paragraph 7.2.3 the headache attributed to spontaneous low CSF pressure. We report on four patients with such a headache and discuss the probable pathophysiology, including results published in the literature. It seems that not the low CSF pressure itself is the cause for the headache but the unphysiological, increased vasodilatation of intracranial and epidural veins. This dilatation of veins also shows up in the typical radiological findings with meningeal contrast enhancement and enlarged epidural veins. A trial with caffeine, theophylline, or indomethacin is recommended; otherwise, the most effective treatment option is an epidural blood patch, which is effective also in the absence of a documented CSF leak. PMID- 15338142 TI - [Doctor and patient: Paul Flechsig and Daniel Paul Schreber]. AB - During his three stays in psychiatric institutions between 1884 and 1911, Daniel Paul Schreber was also twice treated in the psychiatric and neurological hospital of Leipzig University headed by Paul Flechsig. In his "Memoirs of a Nervous Patient," Schreber also described the treatment he had received there, which in his perception seemed to be of crucial importance for his later fate. Research on the Schreber case in the last 25 years, based mainly on the interpretation of his own memoirs, supported the view that in fact Flechsig was to blame to a great extent for the outbreak of Schreber's paranoid schizophrenia in February 1894. This paper aims at forwarding exonerating arguments in favor of Flechsig, concluding that what he did did not differ distinctively from the standard tenets of organic psychiatry taught at that time. Of course this does not change the fact that Schreber did not receive the right kind of treatment. However, it puts Flechsig's approach into the right perspective. PMID- 15338141 TI - [Thrombophilias in patients with ischemic stroke. Indication and calculated costs for evidence-based diagnostics and treatment]. AB - Patients with ischemic stroke are sometimes found to have an underlying inherited (deficiency of protein C, protein S, antithrombin III, activated protein C resistance, prothrombin gene mutation, hyperhomocysteinemia) or acquired thrombophilia (lupus anticoagulant and anticardiolipin antibodies, hyperhomocysteinemia). Patient selection for thrombophilia screening is, therefore, a frequent question in managing patients with ischemic stroke. In this review we discuss patient selection and timing for laboratory tests for thrombophilia screening in stroke patients based on a literature review and we calculated overall costs per year in Germany for testing patients older than 18 years with an ischemic stroke of undetermined cause. As there is a lack of studies comparing anticoagulation with antiplatelet therapy in patients with diagnosed thrombophilia, laboratory screening for thrombophilia even in a selected group of patients with cryptogenic ischemic stroke remains of questionable value at present. An exception appears to be testing for lupus anticoagulant and anticardiolipin antibodies in younger patients with suspected antiphospholipid syndrome (two positive test results necessary), because anticoagulation seems to be superior to aspirin in patients with antiphospholipid syndrome. PMID- 15338143 TI - [Perspectives in cardiological research]. AB - German cardiological research is confronted with increasing difficulties. Clinical research is restricted by regulations, such as the working hours protecting law, the revised version of the legal articles against corruption and acceptance of advantage as well as by many parts of law for the general frame of the university structures. In addition more and more administrative duties are tranferred to doctors engaged in research. Furthermore cardiology is at a disadvantage as only part of the net profits for cardiological services are tranferred to the responsible clinic. Likewise the facilities for cooperation are increasingly restricted, as basic science institutions originally allocated to cardiological research, are now devoted to other subjects and as many pharmaceutical firms have left the country. Cardiology in our country is practically not supported by private research organizations. Research projects are, therefore, predominantly financed by grants from the Bundeministerium fur Bildung und Forschung and by the Deutsche Forschungsgemeinschaft. The financial resources for research in our country are declining and much smaller compared, e. g., to the USA. As a consequence of the shortage of resources not only are the weak projects turned down; it is feared that also the very innovative projects are likewise excluded for entering unknown territory. In periods of financial restrictions the central office and the experts evaluating the projects have a special responsibility, which cannot be met by technical objections, such as e. g., an "unsufficient impact factor". In order to improve the conditions for cardiological research the net profits for cardiological services should be transferred unrestricted to the responsible clinic. The acceptance rate of cardiologcal projects may be increased by more intensive cooperation. At the end, the principle of help by self-help also applies to cardiological research; the British Heart Foundation has developed into an impressive research-supporting organization in similarly difficult times. PMID- 15338144 TI - [Studies on morphogenesis and visualization of the early embryonic heart with regard to the development of conotruncal heart defects]. AB - Most congenital cardiovascular malformations have their origins during early morphogenesis, and some forms of adult-onset cardiovascular disease also arise during embryonic development. Conotruncal heart defects comprise a major category of congenital heart disease and are found in children with a relative high frequency. These defects are associated with a high mortality risk in utero, and after postnatal surgical repair; embryologically they are linked with dextroposed aorta, which is an anomaly of the ventricular outflow tract with malalignment of the great arteries. The etiology and pathogenesis of dextroposed aorta is not known but is thought to be due to abnormal looping and/or incorrect "wedging" of the outflow tract (i.e., wedged positioning of the aorta between the atrioventricular valves) during early heart development. We have studied the morphology and visual development of the embryonic heart in an animal model of dextropsed aorta in a series of experiments to determine possible mechanisms for dextropositioning of the aorta. At this, we have employed besides established methods for analysis of anatomy and pathology (morphological studies, cardiac morphometry, histology, scanning electron microscopy and immunhistochemistry) also new imaging techniques (videocinephotography and time-lapse studies with a digital high-speed video camera, confocal and scanning electron microscopy, optical coherence tomography (OCT) and magnetic resonance microscopy (MRM) for 3D reconstruction of the heart) to achieve a better visualization of normal and pathological changes during heart development. The paper at hand summarizes the results of these studies. PMID- 15338145 TI - [Prediction of outcome in ST elevation myocardial infarction by the extent of ST segment deviation recovery. Which method is best?]. AB - Simple and rapid measures are needed for timely assessment of the quality of reperfusion therapy early after fibrinolysis in acute STEMI. Sum ST segment elevation resolution (sum STR) categorized into the three groups of low risk (complete ST resolution), medium risk (partial ST resolution), and high risk (no ST resolution) has become an established method to predict infarct size, left ventricular function, epicardial vessel patency, and mortality. However, measurement of the sum of ST elevation from all leads of repeated ECG's is time consuming. For routine practice more simple measures are needed. This report summarizes recent findings on direct comparisons between different modes of evaluation of ST segment deviation recovery employed for risk stratification in large-scale mortality trials. With respect to predictive accuracy combined with simplicity, two methods were superior to the conventional model of sum STR: 1) ST segment deviation resolution in only the one ECG lead showing the maximal deviation (single lead STR), and 2) the existing ST segment deviation in the single ECG lead of maximum deviation present 90 or 180 min after start of fibrinolysis (max STE). In multivariate analyses the ST segment deviation recovery models including sum STR were significant independent predictors of short- and long-term mortality. In receiver-operating characteristic (ROC) curves for predicting mortality the analysis of single lead STR and max STE performed better than sum STR. After categorization into risk groups patients are best classified by max STE. With an ECG recorded at 90 min in 2719 patients, the proportion of patients of sum STR, single lead STR, and max STE were 40, 34, and 43% in the low risk groups, and 24, 31, and 25% in the high risk groups. Cardiac mortality rates at 30 days were 2.0, 1.2, and 1.0% in low risk versus 9.6, 10.3, and 12.8% in the high risk groups, respectively. Long-term mortality with a followup of 5 years was best predicted by max STE risk groups. CONCLUSION: Single lead STR and max STE are very simple, inexpensive, non-invasive, and highly reliable measures which provide very strong early prognostic information. The relationship between degree of ST segment deviation recovery and subsequent mortality is remarkably consistent. Both methods perform better than sum STR in predicting mortality. They can be used for very early risk stratification and can form a basis for an individual treatment of patients after fibrinolysis for STEMI within 6 hours of symptom onset. Of the two methods max STE is even simpler to use and has better accuracy in predicting outcome. PMID- 15338146 TI - [Polymorphonuclear neutrophils in myocardial ischemia and reperfusion injury. Influence of coronary intervention?]. AB - Circulating polymorphonuclear neutrophils (PMNs) are known to contribute to the pathophysiology of myocardial ischemia and reperfusion injury. The present study was designed to examine whether the duration of ischemia in myocardial infarction, the type of coronary intervention (angioplasty vs. angioplasty plus stenting) and duration of the procedure itself modulate the inflammatory responses of PMNs. METHODS AND RESULTS: In patients with acute myocardial infarction (AMI; n = 20) neutrophil beta-2-integrin Mac-1 (CD11b/CD18) and L selectin (CD62L) were determined at different time points before and after PTCA or PTCA plus stenting. After PTCA alone but not after PTCA plus stenting a significant rise in Mac-1 (136 +/- 43%) was demonstrated. This elevation decreased after 60 minutes if the duration of the recanalization procedure was shorter than 30 minutes but remained elevated after longer interventions. After delayed intervention a significant and more pronounced increase of Mac-1 (142 +/- 37%) was observed, while early intervention prevented this increase. After PTCA alone or delayed intervention a significant shedding of L-selectin (77 +/- 20%; 77 +/- 23%) was demonstrated. Early intervention or PTCA plus stenting caused no significant changes in L-selectin. CONCLUSIONS: It is concluded that PMN activation is attenuated by early and short intervention as well as by stenting. Induction of PMN activation might contribute to the superior outcome following stenting and early intervention compared to conventional PTCA in particular when performed delayed. PMID- 15338147 TI - [Off-pump versus on-pump coronary artery bypass surgery. Comparison of 270 case matched elderly patients]. AB - INTRODUCTION: Off-pump versus on-pump coronary artery bypass surgery: it still remains a matter of debate which method results in a lower incidence of perioperative morbidity and mortality. This case-matched study evaluates the outcome of elderly patients in both groups. METHODS: All patients aged 75 and older, who underwent CABG from 1998 to 2002, were examined retrospectively. They were matched according to Euroscore and the number of diseased vessels. The Student's t-test and chi-square test were used where appropriate. RESULTS: 270 CABG patients were considered: 135 off-pump and 135 on-pump patients. Mean age was 78.4 +/- 3.1 versus 77.5 +/- 2.9 years, respectively. EuroSCORE was 7.11 +/- 2.3 in both groups; number of distal anastomoses per patient 1.7 +/- 0.74 versus 2.6 +/- 0.63 (p < 0.001), operation time 138 versus 177 minutes (p < 0.001). There were no significant differences in postoperative complications including hospital mortality 3.0 versus 3.7%, renal failure 8.9 versus 12.1% (new onset), acute myocardial infarction 1.5 versus 4.4% and cerebral events 0 versus 1.5%, respectively. The number of transfused packed cells was 2.6 +/- 2.8 versus 4.6 +/ 5.3 (p < 0.001). Intubation time and ICU stay were similar in both groups. CONCLUSION: OPCAB is not associated with a reduction of perioperative mortality and morbidity in patients aged 75 and older. PMID- 15338148 TI - [Piercing and tattoos in patients with congenital heart disease -- is it a problem?]. AB - Piercing and tattooing enjoy widespread popularity in modern society. Patients with congenital heart disease are at elevated risk for infective endocarditis. However, it is not yet known whether piercing and tattooing are dangerous for these patients.A search of the literature provided 10 published cases of infective endocarditis after piercing or tattooing. Affected patients were adolescents or young adults ranging in age from 13 to 30 years (5 male, 5 female). Four of the patients had a known cardiac risk factor for endocarditis (bicuspid aortic valve, postoperative trans-position of the great arteries, postoperative coarctation, postoperative aortic valve stenosis). Piercing preceded endocarditis in 9 cases (4 times mouth, 2 ear, 1 nose, 1 breast, 1 navel), one tattoo. The following agents were isolated: S. aureus in 4 cases, 2 S. epidermidis, 1 Str. viridans, 1 Neisseria mucosa, 1 Haemophilus aphrophilus, 1 Haemophilus parainfluenzae. All patients were treated with antibiotics. Six patients underwent cardiac surgery (5 of them valve replacement). Patients with congenital heart disease constitute less than 1% of the population. Thus, they are clearly overrepresented in the published literature. Epidemiologic conclusions are not possible from these data. However, patients with congenital heart disease and their parents should be strongly advised against piercing and tattooing with regard to the risk of infective endocarditis. PMID- 15338149 TI - [Contrast echocardiography for detection of incomplete rupture of the left ventricle after acute myocardial infarction]. AB - Myocardial rupture is a major complication after acute myocardial infarction. With complete rupture of the free left ventricular wall cardiac tamponade occurs with fatal outcome in most cases. With partial rupture, however, hemorrhage is slower, allowing days or weeks for diagnosis. Survival of these patients strongly depends on early recognition of this complication followed by immediate surgical intervention. Echocardiography is the diagnostic tool of choice to detect myocardial rupture with consecutive hemopericardium but diagnosis remains difficult even if suspected. We describe the case of a patient with inferior infarction who presented with cardiogenic shock, echocardiographic signs of pericardial effusion and abnormal motion and myocardial irregularities of the inferior wall. With Doppler echocardiography no flow across the wall was detected. Left heart contrast echocardiography confirmed the diagnosis of suspected myocardial rupture by clear deliniation of the defect. Immediate surgical repair was successfully performed in this patient with favorable long term outcome. Thus, echocardiography early after acute myocardial infarction is useful in detecting subsequent complications and the use of contrast echocardiography should be considered in suspected myocardial rupture. PMID- 15338150 TI - [Reversible electrocardiographic changes in hypothermia]. AB - A homeless man with accidental hypothermia showed massive ECG changes on hospital admission. Including sinus bradycardia, AV-block 1 degree, widened QRS complex with Osborne waves and QT prolongation. These changes were slowly but completely reversible after surface rewarming. PMID- 15338151 TI - [Guidelines: acute coronary syndrome (ACS)]. PMID- 15338153 TI - [Health status and health resources]. PMID- 15338154 TI - [Influence of pain perception, morbidity and mood on functional impairment in elderly chronic pain patients]. AB - The purpose of the study was to address the impact of pain perception, morbidity and mood on functional impairment in elderly chronic pain patients. Multimorbid pain patients beyond the age of 65 in two geriatric hospitals (n = 84), a pain clinic (n = 60) and three general practices (n = 117) provided information about pain perception, comorbidity, additional symptoms and mood by means of the "Structured Pain Interview for Geriatric Patients", the "Cumulative Illness Rating Scale" and a list of symptoms. Data analysis relied on stepwise multiple regression with variables of pain perception entered in the first step, of morbidity entered in the second step and of mood entered in the third step. Although patients believe that pain is the main reason for their functional impairment (71.3%), the data do not support this assumption. Increasing morbidity and bad mood have more impact to reduced functional performance than the pain perception. Our results support the recommendation that a multimodal program should be offered to even multimorbid and older people with chronic pain in order to achieve a maximum of functional rehabilitation. PMID- 15338155 TI - [Psychotherapy with the elderly in outpatient settings: are elderly patients the easier ones?]. AB - Elderly patient's preconditions for outpatient psychotherapy are compared with regard to symptom levels, psychological and bodily functioning, and interpersonal problems with groups of younger patients. Based on the Age and Disorder-Specific Model of Psychotherapy (ADS-MP) we predicted that the group of older patients exhibits more favorable preconditions for psychotherapy as well as a shorter duration of treatments than younger groups. A total of 169 outpatients from an university clinic were divided into four groups and compared. Rates of mental disorders did not differ between the groups for all frequent disorders. The group of elderly patients did not show a difference in symptom levels although their psychological functioning was better than in the comparison groups. Two of eight interpersonal problems showed more favorable initial values for the elderly. The number of necessary therapy hours up to the treatment conclusion was significantly smaller in the older group (on average 21 hours) than with young to middle-aged patients groups. The results point to the fact that the psychological preconditions for psychotherapy in old age are good and indicate promising directions for the development of age-appropriate interventions. PMID- 15338156 TI - [Substance abuse in middle and old age -- general abuse]. AB - The problems of drug-addicted middle- and older-aged people have been disregarded for a long time. However, results show a risky consumption of illegal drugs in the 18- to 59-year-old German population. The following paper will analyse the substance abuse of alcohol, sedatives, cannabis, stimulants, cocaine, opiates and other drugs not described more closely (all of which lead to health problems) using the Interdisciplinary Longitudinal Study of Adulthood (ILSE). Data collection is based on two cohorts (1930-1932 and 1950-1952) at two measurement points (1. MMP: N = 1006, 2. MMP: N = 900) in centres of investigation in Heidelberg and Leipzig. The "Substance Abuse Section" of the Structured Clinical Interview for DSM-III-R (SCID) was used for the evaluation. The abuse of alcohol can be registered as the focal point. Men consume more alcohol, while women tend to take sedatives, stimulants, opiates and other substances more often. The East West comparison shows the importance of accessibility. At both measurement points, abuse of more substance groups was determined in the Heidelberg centre. As for as the abuse rates are concerned, a reduction of all the substance groups can be registered for the second measurement point. PMID- 15338157 TI - [Active ageing, decreasing capability, social disadvantage and age stereotypes: a contribution to resilience and vulnerability in older adulthood]. AB - The starting-point of this contribution is the concept of active ageing as it has been propagated by the World Health Organization since the 1990s. Relationships between subjective experience of the potentials and barriers involved in leading a productive and responsible life, decreases in capability, small financial resources, and social isolation are analysed in a total sample of 1,275 participants. Based on this analysis, two subsamples are selected: a first characterised by a high level of risk, i.e. low opportunities for active ageing, and a second characterised by a low level of risk, i.e. high opportunities for active ageing. Four age stereotypes are analysed as predictors of experienced potentials and barriers in the total sample and the two subsamples. Results suggest varying significance of age stereotypes for constellations of resilience and vulnerability. From an age-stratification perspective, it is argued that results reflect a reservation against old age in our culture. The specific contribution of age stereotypes accentuating gains and chances versus losses and risks to understanding resilience and vulnerability are discussed from a social constructionist perspective. PMID- 15338158 TI - [Somatoform complaints in the elderly. Prevalence and associations with personality variables, career, and family]. AB - The aim of this study is to determine the prevalence of clinically relevant somatoform complaints in a representative sample of the elderly. For the first time, the job-related development of the elderly is analysed in relation to somatoform complaints. Further, personality variables and family development are examined. The data basis of the evaluation is a partial sample (n = 1002) of the cohort study "Interdisziplinare Langsschnittstudie des Erwachsenenalters" (ILSE). The assessment included a half-structured interview on the development of work and family, a detailed clinical examination, a psychiatric screening (SCID) and the neuroticism and extraversion scales of the personality test NEO-FFI. A total of 29 participants (2.9%) with clinically relevant somatoform complaints were identified, 20 females and 9 males. They were significantly less satisfied and less involved in their careers than the rest of the sample. They evaluated their partnership as significantly worse, yet the amount of experienced distress did not differ. The index group scored much higher on scales for neuroticism than the other, whereas they scored significantly lower on scales for extraversion. PMID- 15338159 TI - [Frontotemporal dementia: specific problems for caregivers]. AB - Frontotemporal dementia is a relatively rare neurodegenerative disease. In the majority of cases the onset is at a presenile age, and changes of behavior as well as alterations of personality and social conduct are predominant symptoms whereas cognitive deficits are mild. Therefore interventions for caregivers including self-help groups and educational programs which are usually tailored to Alzheimer's disease often do not meet the needs of caregivers of patients with FTD. The present paper reviews the clinical features of FTD, outlines the specific problems and burdens that caregivers are faced with, and describes novel interventions for this group of family caregivers. PMID- 15338160 TI - [Gerontopsychiatry and nursing science. Similarities and differences]. AB - In the following article, two of the increasingly important public health scientific disciplines of care for the elderly are compared under systematic aspects - gerontopsychiatry and nursing science - with the aim of determining the similarities and differences of both disciplines. There follows a discussion of social challenges facing the area of caring for the elderly suffering from psychiatric illnesses and the responsibilities which both disciplines have to cope with in the future. The clearing of the numerous existent geronto psychiatric deficits in health care in Germany seems to be the main challenge. PMID- 15338161 TI - [The Barthel Index in geriatrics. A context analysis for the Hamburg Classification Manual]. AB - The Barthel Index (BI) is firmly established as an assessment instrument in geriatrics. It is a proven, clear and easy-to-use instrument for the recording of basic daily functions. However it is increasingly finding new applications beyond its original, clinically orientated use. It has been applied as a quality parameter, as an instrument for the management of service delivery provision and as an instrument to record treatment efforts relevant to care or costs. This study considers the basic suitability prerequisites of the Barthel Index for these applications.With the Hamburg Classification Manual for the BI, German geriatrics has made a contribution to the standardized operationalization of the items and to the standardized evaluation of aids. An evaluation was performed on a total of 5262 Barthel classifications at seven geriatric hospitals according to existing classification practice by comparison with 5483 classifications after introduction of the Hamburg Classification Manual. No significant differences were found in Barthel Index total scores either on admission or on discharge. With respect to further applications it is essential to be aware that the usability of the total score (including its changes over the course of time) is limited because of the ordinal scaling of the BI. Studies have been carried out which show how important this is, although they have so far received little attention. As a grading criterion the BI takes account not only of the "functional status" but also of the "extent of support effort". This can lead to positive changes in one focus -- especially with the use of aids -- without associated improvements in the other focus. Whether the BI in this form is meaningful for a specific application must be tested separately for each individual context. There is no justification for assuming that the BI has general validity irrespective of application. The results of a systematic literature survey on the testing quality criteria of the BI indicate an astonishingly generous approach to the question of the validity of the BI. Contrary to widespread opinion, cognitive-psychological components do influence the classification result of the BI. As an addition to the Hamburg Classification Manual we recommend that the extent of this influencing factor should also be more clearly operationalized and that "stimulation required" for the carrying out of an activity be included in the evaluation equivalent to the factor "supervision required" already introduced by Barthel and Mahoney. The BI has shown itself to be an efficient but nonetheless multidimensional global parameter in clinical practice, whose meaningfulness is on the level of the individual item and whose validity, particularly as total score and course parameter, needs to be proven for each new application. The Hamburg Classification Manual, as a standardized and consensus-based operationalization of the BI, provides an important basis for this. PMID- 15338166 TI - Ameliorative effect of IDS 30, a stinging nettle leaf extract, on chronic colitis. AB - BACKGROUND AND AIMS: Anti-TNF-alpha antibodies are very effective in the treatment of acute Crohn's disease, but are limited by the decline of their effectiveness after repeated applications. The stinging nettle leaf extract, IDS 30, is an adjuvant remedy in rheumatic diseases dependent on a cytokine suppressive effect. We investigated the effect of IDS 30 on disease activity of murine colitis in different models. METHODS: C3H.IL-10-/- and BALB/c mice with colitis induced by dextran sodium sulphate (DSS) were treated with either IDS 30 or water. Mice were monitored for clinical signs of colitis. Inflammation was scored histologically, and faecal IL-1beta and mucosal cytokines were measured by ELISA. Mononuclear cell proliferation of spleen and Peyer's patches were quantified by 3H-thymidine. RESULTS: Mice with chronic DSS colitis or IL-10-/- mice treated with IDS 30 clinically and histologically revealed significantly (p < 0.05) fewer signs of colitis than untreated animals. Furthermore, faecal IL 1beta and mucosal TNF-alpha concentrations were significantly lower (p < 0.05) in treated mice. Mononuclear cell proliferation after stimulation with lipopolysaccharide was significantly (p < 0.001) reduced in mice treated with IDS 30. CONCLUSIONS: The long-term use of IDS 30 is effective in the prevention of chronic murine colitis. This effect seems to be due to a decrease in the Th1 response and may be a new therapeutic option for prolonging remission in inflammatory bowel disease. PMID- 15338167 TI - A novel approach for perirectal tumours: the perianal intersphincteric excision. AB - INTRODUCTION: Several procedures have been described for the excision of perirectal tumours. Among them are the Kraske and York Mason operations, and the transanal approach, which may cause anal incontinence and rectal bleeding. A novel technique, i.e. the perianal intersphincteric excision (PIE), is reported here with the aim of minimising such postoperative complications, as it avoids both stretching the sphincters and endorectal wounding. CASE REPORT: The procedure has been carried out successfully in two patients: a woman with a low grade rectal leiomyosarcoma and a man with a perirectal schwannoma. No postoperative complications were observed, and no recurrence was detected after 36 and 8 months respectively. CONCLUSION: In conclusion, PIE seems to be a safe and effective procedure for the management of perirectal masses. PMID- 15338168 TI - Gigantic gastrointestinal stromal tumor in the pelvis. PMID- 15338169 TI - Congenital duodenal obstruction: does prenatal diagnosis improve the outcome? AB - Prenatal diagnosis of congenital duodenal obstruction (CDO) provides information about associated anomalies; helps plan the delivery, resuscitation, and neonatal surgery; and allows for appropriate family counseling. This report compares the outcomes of two groups of newborns: one with prenatal diagnosis of CDO (group I) and the other without (group II). Charts of the 23 newborns with CDO admitted to the Hospital of UNICAMP between 1993 and 2001 were retrospectively reviewed. Ten (44%) newborns had prenatal diagnosis of CDO. Among group I patients, the postnatal diagnosis was confirmed on the 1st day of life, whereas patients without prenatal diagnosis (group II) had the diagnosis of CDO confirmed at a mean age of 5.7 days ( p=0.004). The mean ages at surgery, at total oral feeding, and at hospital discharge were also statistically lower among infants with prenatal diagnosis, and more complications occurred in group II patients. The earlier care could explain the statistically lower morbidity for patients with prenatal diagnosis, since they were able to undergo further investigation and surgical repair before any impairment to their clinical status could take place. We believe that prenatal diagnosis of CDO, associated with earlier surgery and adequate postoperative support, can provide lower morbidity, decrease the hospitalization period, and, therefore, decrease its costs to the state and to society. PMID- 15338170 TI - Pseudoaneurysms of the brachial artery following venipuncture in infants. AB - Pediatric vascular injuries are increasing in frequency and represent a challenging problem in pediatric surgical practice. Increased survival of low birth weight infants and advances in invasive diagnostic procedures have resulted in a dramatic increase in the number of these injuries. Formation of pseudoaneurysm of the brachial artery in infants is a very rare complication of venipuncture, with only two cases reported in the literature. We report three cases of brachial artery pseudoaneurysm in infants following venipuncture who were operated upon in our institution, aged 43-64 days at the time of operation. The period from the injury to the operation ranged from 25 to 42 days. All three infants were referred from different institutions. In two infants, the pseudoaneurysms and the involved part of the artery were resected, and arterial continuity was restored with an end-to-end anastomosis; in the other infant, reconstruction was done using a venous interposition graft. All three infants were diagnosed with duplex ultrasonography, and the child requiring a more complex reconstructive procedure was also evaluated with helical contrast computed tomography. Brachial artery pseudoaneurysms are a rare but possible complication of multiple venipuncture in infants. Early diagnosis and microvascular reconstruction are key points in managing these injuries. PMID- 15338171 TI - Role of nitric oxide and cyclooxygenase pathway in lipopolysaccharide-induced intussusception. AB - The etiology of intussusception (IN) remains largely obscure. In lipopolysaccharide (LPS)-induced IN, an experimental model in mice, IN is considered to be the consequence of altered intestinal motility as a result of increased nitric oxide (NO) along various inflammatory mediators. These could be decreased via cyclooxygenase (COX) inhibition by indomethacin. N-omega-nitro- L arginine methyl ester ( L-NAME) inhibits nitric oxide synthase (NOS) and NO production. Indomethacin is known to prevent IN; however, the reason is unknown. In this study we aimed to determine the role of NO, the effects of inhibition of its production by L-NAME and indomethacin, and whether preventive effects of indomethacin on LPS-induced IN were related to NO inhibition. A total of 113 mice were divided into seven groups. In the control group ( n=6), no procedure was done. In the sham group ( n=6), 1 ml saline was given; in the indomethacin group ( n=6), 10 mg/kg of indomethacin was given; and in the LPS group ( n=30), 12 mg/kg of LPS was administered intraperitoneally (IP). In the LPS+indomethacin group ( n=32), 10 mg/kg of indomethacin was administered IP simultaneously with 12 mg/kg of LPS. In the L-NAME group ( n=6), 20 mg/kg of L-NAME was administered subcutaneously. In LPS+L-NAME group ( n=27), 20 mg/kg of L-NAME was administered subcutaneously with 12 mg/kg of LPS IP. All animals were laparotomized 6 h following injections. Existence of IN was noted and blood specimens were obtained. NO was quantified by measurements of nitrite and nitrate, obtaining a total of NO metabolites (NOx). The results were compared using the Mann-Whitney U test and Spearman correlation test. A value of p<0.05 was considered significant. A total of 17 mice (one in control, 10 in LPS, four in LPS+indomethacin, and two in LPS+L-NAME groups) were excluded from the study because of death or insufficient blood collection. LPS (12 mg/kg, IP) induced IN at a rate of 30% ( n=6) in the LPS group. Mean NOx levels were statistically higher in the LPS group (186.67+/-20.06) compared with other groups ( p<0.05). Mean NOx levels were significantly higher in the group of mice with IN than in those without in the LPS group of this study (295.46+/-16.42, 140.05+/-15.44, respectively, p<0.05). The mean NOx levels were statistically lower in the LPS+ L-NAME(23.94+/-3.39) group than the LPS+indomethacin (106.77+/-24.54) group, with no IN detected in neither of these two groups. Increased NOx levels induced by LPS correlated well with the occurrence of IN, and decreasing these levels via COX inhibition by indomethacin or NOS inhibition by L-NAME totally prevented IN from forming in this study. By these observations, it could be concluded that NO is probably involved in the pathophysiology of IN in this experimental model of LPS-induced IN. PMID- 15338172 TI - Esophageal atresia and other visceral anomalies in a modified Adriamycin rat model and their correlations with amniotic fluid volume variations. AB - The Adriamycin rat model (ARM) has been used to produce visceral malformations in fetuses to explain the mechanisms of foregut division. The models vary in the dosage of Adriamycin (ADR) and in the number of applications. Our study of a modified ARM using 2.2 mg/kg of ADR for 2 days only, intraperitoneally in pregnant rats, is presented. A total of 81 fetuses were obtained with this model from the ADR group, 74 (91%) alive. Uretero-hydronephrosis (UHN) was observed in 70 fetuses (95%), esophageal atresia (EA) in 68 (92%), duodenal atresia (DA) in 68 (92%), bladder hypoplasia (BH) in 67 (90%), plus other malformations. In evaluating amniotic fluid (AF) volume of the fetuses with EA with tracheo esophageal fistula (TEF) (group I) and EA without TEF (group II), both associated with bilateral UHN when compared with the control group (group III), groups I and II showed higher AF volume in groups I and II than the control group (group III) did ( p=0.0001). In conclusion, ARM was adequate to produce EA and other visceral malformations. The use of ADR in a higher dosage for a shorter period of time produced better results than those presented in previous literature. The increase of AF volume obtained in fetuses presenting EA plus bilateral UHN strongly suggests, despite ureteral dilatation (urinary obstruction), that a malformed communication may exist between the urinary system and the amniotic cavity, permitting the existence of polyhydramnios that is due to digestive obstruction such as EA and DA. PMID- 15338173 TI - The effect on the intestines of continuous release of methylene blue from a drug delivery system: an experimental study in a chick embryo gastroschisis model. AB - Increased small bowel nitric oxide synthase (NOS) activity has been suspected as a cause of postnatal intestinal dysmotility in gastroschisis. The effect of continuous delivery of methylene blue loaded polymer (MBLP) hydroxy-propyl methyl cellulose-ethyl cellulose (HPEC-MC) and daily injection of methylene blue (MB) on the intestinal damage (ID) was evaluated using a chick embryo gastroschisis model. Fourteen-day-old fertilized chick eggs were divided into five groups. In the control (C) group, no intervention was performed. In the sham (S) group, the allantoic and amniotic membranes were opened to create a common cavity that resembles the amniotic cavity in human. In the gastroschisis only (GO) group, a defect in the abdominal wall of the embryo was made, and intestinal loops were exteriorized following connection of amniotic and allantoic cavities. In the gastroschisis plus methylene blue (G+MB) group, gastroschisis was created and MB administered into the amnioallantoic cavity (AAC) by daily injections for 5 days. In the gastroschisis plus methylene blue loaded polymer (G+MBLP) group, MBLP was placed into AAC after gastroschisis was created. At the end of the 19th day of incubation, intestinal morphological changes were investigated macroscopically and microscopically. Although the survival rates were decreased in the chick embryos with creation of gastroschisis compared with C and S groups ( p<0.001), the survival rates were increased in G+MBLP group (76.92%) when compared with the GO group (41%) ( p<0.001). Because of multiple intervention of embryos, higher mortality was observed in the G-MB group (75.61%). Macroscopic and microscopic scores of ID and mean intestinal wall thickness were significantly higher in the GO group when compared with C, S, G+MB, and G+MBLP groups ( p<0.001). The mean score of intestinal ganglia morphology was significantly increased and the total number of ganglion cells was significantly decreased in the GO group when compared with C, S, G+MB, and G+MBLP groups ( p<0.001). It is possible to decrease intrauterine intestinal morphological changes in gastroschisis by inhibiting NOS. As a first preliminary study, we believe that use of MBLP may be an alternative for fetal treatment by eliminating the harmful effects of multiple interventions or amniotic fluid exchanges. PMID- 15338174 TI - The effects of orchidopexy and orchidectomy on testes of rats subjected to ipsilateral abdominal testis with vas deferens obstruction. AB - An experimental study was planned to evaluate and compare the effects of orchidopexy and orchidectomy on the testes of rats subjected to unilateral abdominal testis with vas deferens obstruction. Four groups were established. Rats in the control group underwent a sham procedure. While the testis was maintained in the abdomen with the vas deferens ligated for 8 weeks in group 2, rats in groups 3 and 4 underwent orchidopexy or orchidectomy after 4 weeks. Remaining testes were harvested at the end of the 8-week period. Testis and body weights were obtained during harvest. Samples were evaluated through DNA flow cytometry, and percentages of haploid cells were determined. Groups were compared through unpaired t-test, and p-values less than 0.05 were considered significant. All three treatments had decreased testis weight over body weight values of ipsilateral testes. Ipsilateral orchidectomy increased the value among contralateral testes. However, none of the groups had a contralateral testicular value less than the sham-operated group. All three treatments decreased the percentage of haploid cells among ipsilateral testes, but only an abdominal testis was associated with a decrease in the percentage of haploid cells among contralateral testes. Maintaining a testis with an obstructed vas deferens in the abdomen for 8 weeks damages both ipsilateral and contralateral testes. Orchidopexy, while showing amelioration of the ipsilateral testis, spares the contralateral testis as well as orchidectomy. Orchidopexy for an undescended testis with vas deferens obstruction is a rational approach. PMID- 15338175 TI - High scrotal (Bianchi) single-incision orchidopexy: a "tailored" approach to the palpable undescended testis. AB - Our aim was to evaluate the utility of the high scrotal orchidopexy (Bianchi) approach for palpable undescended testis (UDT) and to assess long-term follow-up. We reviewed the records of orchidopexies performed between 1999 and 2002. The patients were then categorized by intraoperative exam under anesthesia as to whether their testes were palpable or nonpalpable. All palpable UDT that were initially thought to be amenable to a single high scrotal approach (Bianchi) were then reviewed. These cases were then analyzed to assess the impact of patient age, initial location of the testis, and prior inguinal/scrotal surgery with respect to the necessity to convert to a standard two-incision technique, and to analyze success and complications at 6-12-week and 1-year follow-up. Two hundred and nineteen orchidopexies were performed on 204 patients over this 4-year period. There were 178 testes palpable, and the transscrotal approach was used in 85 patients (100 orchidopexies). The preoperative positions of the testes that were thought to be amenable to Bianchi technique included the following: gliding (19), secondary trapped (25), superficial inguinal pouch (42), and location within the inguinal canal (2), while the remaining 12 testes were ectopic. Six patients required conversion to a traditional inguinal approach because of insufficient cord length via the single incision to allow the testis to lie in the scrotum. All patent processes vaginalis were ligated via the scrotal incision, regardless of their size. All patients, except for one who had a testis in the superficial inguinal pouch, had palpable testes of stable size and in a dependent position at 6-12-week follow-up. Of the 62 children who returned for 1 year follow-up, all had findings identical to those at their initial 6-week visits, with no atrophy or secondary reascent. Postoperative complications included transient postoperative scrotal hematoma in a single patient. The single failure underwent a successful two-incision orchidopexy for secondary reascent and a resultant trapped testis. Children with primary palpable undescended, gliding, or trapped testes can be managed successfully through the transscrotal route in the majority of cases. With use of a tailored approach to the palpable UDT, an additional groin incision is necessary only for a minority of appropriately selected cases. PMID- 15338176 TI - Posterior urethral diverticulum after laparoscopic-assisted repair of high-type anorectal malformation in a male patient: surgical treatment and prevention. AB - Currently, laparoscopic-assisted colon pull-through (LACPT) is the treatment of choice for male patients with high-type imperforate anus and rectourethral fistula. Since laparoscopy was introduced for treating this condition, reports concerning post-LACPT complications are rare. Here we discuss the case of a boy, now 3.5 years old, born at 37 weeks' gestation weighing 2,300 g, who was diagnosed with rectobulbar urethral fistula (RUF) at birth. LACPT was performed when the boy was 11 months old and weighed 7.2 kg. No intraoperative complications occurred, and the initial post-LACPT course was uneventful. When he was 2 years old, he developed dysuria requiring urethral catheterization. Diagnostic radiology confirmed a large cystic mass behind the bladder, suggestive of a posterior urethral diverticulum (PUD). Histopathology of the excised mucosa of the cyst showed colonic mucosa, confirming that the cyst was indeed an enlarged residual RUF. We discuss our treatment and our approach to prevention. PMID- 15338177 TI - Development in harmony. PMID- 15338178 TI - The significance of skull fracture in mild head trauma differs between children and adults. AB - OBJECTIVE: The objective was to determine whether the age of patients with mild head injury and skull fracture influences the level of risk for acute intracranial injuries. METHOD: A study was conducted of 156 patients with skull fracture, 60 children (aged <14 years) and 96 adults, detected among 5,097 consecutive patients with mild head injury (Glasgow Coma Scale [GCS] score of 15 14 points) arriving at the Emergency Department of a Level I University Hospital Trauma Center during 1998. Acute intracranial injuries were defined as traumatic brain injuries identified by cranial computed tomography scan, excluding pneumocephalus. RESULTS: Compared with the children, this risk of intracranial injury was 13 times greater in the adults aged 14-54 years and 16 times greater in the over-54-year-olds. Besides age over 14 years (p<0.0001), compound skull fracture (p<0.001), and a GCS score of 14 (p<0.001) were factors significantly associated with intracranial injury in the logistic regression analysis. CONCLUSIONS: Skull fracture in mild head injury implies a greater risk of intracranial injury in adults than in children. PMID- 15338179 TI - Ultrastructure of the cerebrospinal fluid outflow along the optic nerve into the lymphatic system. AB - OBJECT: To explain the spontaneous CSF outflow into the orbit, the ultrastructure of the perineural meningeal layers at the distal and the proximal portions of the optic nerve were compared. METHODS: Ten cats were perfusion fixated and the orbital content removed for transmission and scanning electron microscopy. In five animals a 60-min cisternal infusion of contrast medium at low intracranial pressure was performed before perfusion fixation. RESULTS: In the contrast infused animals it was possible to demonstrate the leakage of contrast medium in the distal portion of the optic nerve sheath (ONS) from the subarachnoid space (SAS) into the orbit and find it in the conjunctival lymphatics. Electron microscopy revealed that in the distal portion of the ONS the neurothelial layers are significantly thinner, some consisting of only one layer. Pore-like openings in the neurothelial covering are seen in the distal portion. Excavations of the SAS are far more numerous in the distal portion of the ONS. The excavations reach the neurothelial layer. Intracellular and extracellular filaments are more numerous in the distal portion of the ONS. There is no significant difference in the dura mater between the distal and proximal ONS. The results show the existence of an arachnoid window area in the distal portion of the ONS. It is characterised by a continuous, but thinned neurothelial barrier layer, with few pore-like openings. CONCLUSIONS: The main differences between distal and proximal ONS are a thinned neurothelial barrier layer and an increased number of intercellular filaments and pore-like openings. The findings explain the lymphatic CSF outflow pathway along the optic nerve. PMID- 15338180 TI - The use of neoadjuvant chemotherapy to achieve complete surgical resection in recurring supratentorial anaplastic ependymoma. AB - INTRODUCTION: Ependymoma is a central nervous system neoplasm that is often managed with surgery and radiotherapy. The benefits of chemotherapy in treating this tumor remain poorly defined. CASE REPORT: The use of a platinum-based chemotherapy as a neoadjuvant treatment to permit complete surgical resection of a supratentorial anaplastic ependymoma recurring for the third time is described. DISCUSSION: This paper also discusses the possible use of chemotherapy as a strategy that may allow tumor shrinkage and ease tumor resection in giant recurring ependymomas. PMID- 15338181 TI - Entrainment of leech swimming activity by the ventral stretch receptor. AB - Rhythmic animal movements originate in CNS oscillator circuits; however, sensory inputs play an important role in shaping motor output. Our recent studies demonstrated that leeches with severed nerve cords swim with excellent coordination between the two ends, indicating that sensory inputs are sufficient for maintaining intersegmental coordination. In this study, we examined the neuronal substrates that underlie intersegmental coordination via sensory mechanisms. Among the identified sensory neurons in the leech, we found the ventral stretch receptor (VSR) to be the best candidate for our study because of its sensitivity to tension in longitudinal muscle. Our experiments demonstrate that (1) the membrane potential of the VSR is depolarized during swimming and oscillates with an amplitude of 1.5-5.0 mV, (2) rhythmic currents injected into the VSR can entrain ongoing swimming over a large frequency range (0.9-1.8 Hz), and (3) large current pulses injected into the VSR shift the phase of the swimming rhythm. These results suggest that VSRs play an important role in generating and modulating the swim rhythm. We propose that coordinated swimming in leech preparations with severed nerve cords results from mutual entrainment between the two ends of the leech mediated by stretch receptors. PMID- 15338182 TI - [Introduction to the topic of functional assessment of sporadic visual field defects]. AB - We present the new recommendation released by the Commission of the German Ophthalmologic Society on evaluation of sporadic visual field defects, which is based on the principle of the Estermann field test. The purpose of the recommendation is to ensure quality assurance and legal certainty in assessment of sporadic visual field defects by field evaluation with a point system. PMID- 15338183 TI - [Proposed scoring system for assessment of functional impairment due to visual field defects]. AB - INTRODUCTION: The purpose of the study is the development of a scoring system for the assessment of functional impairment due to visual field defects, based on the Esterman grid, but adapted to the existing system in the Federal Republic of Germany. METHODS: The score areas and the evaluation rules were altered iteratively, until the functional score by the new system matched as much as possible the score obtained from the existing system of geometrical defect patterns. RESULTS: The visual field is divided into 100 score areas, which are smaller in the visual field center and in its lower region, respectively. The areas inside the reference isopter are counted and taken as total function in percent. Areas with monocular perception are rated with 3/4 points. Functional impairment is defined as the difference of total function to 100%. For 9 of 20 defect patterns, the functional scores are equal. Another six patterns differ by 1-4%. The remaining 5 of 20 patterns differ by 6-8% in either direction. CONCLUSION: The new system is easier to apply, more consistent, and covers irregular defect patterns. The level of functional impairment is nearly identical with both systems. A transition to the scoring system would considerably increase the certitude and the quality of assessment especially in difficult cases. PMID- 15338184 TI - [Quality management in the ophthalmologist's practice]. AB - Since 1 January 2004, new health system rules have become law in Germany. All ophthalmologists who practice in the German national health system are obliged to implement an internal quality management system. However, for economic reasons it is necessary for every physician as the owner of a medium-sized business to introduce a quality management system to survive in the healthcare market. The rules in the German health system are changing rapidly and physicians have to react to these new situations. The competition on the healthcare market has begun and will become much harder in the future. Quality management is the tool to successfully handle this challenge. PMID- 15338185 TI - [Acute worsening of vision in an HIV-positive female patient with a helper cell level of over 200/microl]. PMID- 15338186 TI - [Unilateral, segmental corneal decompensation. Suspicion of keratitis herpetica]. PMID- 15338187 TI - Improvement of image quality of multislice spiral CT scans of the head and neck region using a raw data-based multidimensional adaptive filtering (MAF) technique. AB - The purpose was to evaluate the potential of the multidimensional adaptive filtering (MAF) technique by investigating its effects on image noise and image quality in multislice spiral CT (MSCT) examinations of the head and neck region. Fifty patients with head and neck tumors were examined using MSCT with a high resolution protocol. Reconstructions were performed using dedicated reconstruction software with a standard algorithm both without and with MAF using different modification. In all reconstructions, we measured the noise in seven different anatomical structures. The image quality and image noise were rated on a five-point scale. There was a significant (P<0.05) reduction in mean pixel noise in the reconstructions using MAF in comparison to the standard reconstructions, but there was no significant difference between the different modification fractions. With MAF the mean reduction in noise level was 60%, depending upon body shape and anatomical region. Independently from the used modification fraction, MAF led to a significant (P<0.05) improvement of image quality. In direct comparison of the different filter strength, the optimal image quality was achieved in the investigations with 15% MAF. The use of MAF facilitates the distinction of anatomical and pathological structures from artifacts in the supraclavicular fossae and the upper mediastinum, whereas the image quality of the upper portions of the neck remained unchanged. MAF improved image quality by reducing the noise level and removing noise structures without loss of image sharpness. This technique offers new perspectives to reduce the patient dose. PMID- 15338188 TI - Agrobacterium-mediated transformation of European chestnut embryogenic cultures. AB - An innovative and efficient genetic transformation protocol for European chestnut is described in which embryogenic cultures are used as the target material. When somatic embryos at the globular or early-torpedo stages were cocultured for 4 days with Agrobacterium tumefaciens strain EHA105 harbouring the pUbiGUSINT plasmid containing marker genes, a transformation efficiency of 25% was recorded. Murashige and Skoog culture medium containing 150 mg/l of kanamycin was used as the selection medium. The addition of acetosyringone was detrimental to the transformation efficiency. Transformation was confirmed by a histochemical beta glucuronidase (GUS ) assay, PCR and Southern blot analyses for the uidA (GUS) and nptII (neomycin phosphotransferase II) genes. At present, 93 GUS-positive chestnut embryogenic lines are being maintained in culture. Low germination rates (6.3%) were recorded for the transformed somatic embryos. The presence of the transferred genes in leaves and shoots derived from the germinated embryos was also verified by the GUS assay and PCR analysis. PMID- 15338189 TI - Antiphospholipid antibodies in children with systemic lupus erythematosus: a prospective study in northern India. AB - Twenty-seven children with systemic lupus erythematosus (SLE) were tested for antiphospholipid antibodies (APLA), i.e. lupus anticoagulant and immunoglobulin (Ig)G or IgM anticardiolipin antibodies (ACLA), with beta-2 glycoprotein I as cofactor, in a single-centre, prospective study over 2 years. Eighteen patients (67%) tested positively for one or the other APLA during the course of the study. Twelve children (44%) tested positively for IgG ACLA and ten (37%) for IgM ACLA, whereas eight (30%) were positive for lupus anticoagulant. In two patients with thrombosis, IgG anticardiolipin positivity was seen to be variable. Unlike the results of most other reports in the literature, lupus anticoagulant positivity was not consistently associated with thrombosis. A majority of the children (83%) tested positively for ACLA during disease activity. Immunoglobulin G and IgM ACLA positivity did not correlate significantly with disease status. The results of this prospective study would indicate that, though frequently present, APLA may be unable to be predictive of disease behaviour in children with SLE. PMID- 15338190 TI - Acute erythroleukemia in a rheumatoid arthritis patient during low-dose methotrexate therapy. AB - Acute leukemia is uncommonly seen with rheumatoid arthritis during or following treatment with low-dose methotrexate, a safe and effective treatment for the arthritic condition. We describe here a 68-year-old woman with rheumatoid arthritis who developed acute erythroleukemia during low-dose methotrexate therapy (total dose 1702.5 mg). This may be the first such case reported in the literature. PMID- 15338191 TI - Factors and signals that govern the migration of dendritic cells via lymphatics: recent advances. AB - Dendritic cell (DC) migration from peripheral organs to lymph nodes plays a key role in initiating immune responses, whether migratory DCs bring antigen in tow to lymph nodes or position themselves to capture antigen that drains into the lymph node. CCR7 prominently controls DC migration into afferent lymphatic vessels and the positioning of DCs within the lymph node. Expression of CCR7 is not sufficient for function, as its function is positively regulated by a variety of other extracellular triggers. At least one of these triggers, synthesis and secretion of PGE(2), is brought on by the activation of p38 MAP kinase. The MAP kinase pathway has been well studied in DCs and exhibits a complex regulatory role in which the activation of different MAP kinase members leads to biologically distinct outcomes that are dependent upon stage of differentiation at the time of activation as well as the duration of signaling. Almost all of our knowledge of how DCs mature and ultimately mobilize to lymph nodes comes from studies in which DC migration is probed in the context of immune activation and priming. A reasonable body of evidence has gathered to suggest that many molecular events important for DC migration in this context do not affect accumulation of DCs in lymph nodes in the steady state, but mediators that interface with the signaling adaptor DAP-12 may play key roles in the steady state. It may thus become possible to devise approaches to modulate DC mobilization in the context of inflammation without affecting the traffic of DCs during more quiescent conditions. Considering the finely tuned regulation of DC maturation, migration, and cytokine production, with the realization that these phenotypes can be mutually exclusive, manipulation of DC migration in the clinic will be a challenging, albeit feasible, task. PMID- 15338192 TI - Pharmacokinetics and pharmacodynamics of 17-demethoxy 17-[[(2 dimethylamino)ethyl]amino]geldanamycin (17DMAG, NSC 707545) in C.B-17 SCID mice bearing MDA-MB-231 human breast cancer xenografts. AB - PURPOSE: 17-demethoxy 17-[[(2-dimethylamino)ethyl]amino]geldanamycin (17DMAG, NSC 707545) is a water-soluble analogue of 17-(allylamino)-17-demethoxygeldanamycin (17AAG), a compound currently in clinical trials. These preclinical studies: (1) characterized 17DMAG concentrations in plasma, normal tissues, and tumor after i.v. delivery to mice; and (2) correlated tumor and normal tissue 17DMAG concentrations with alterations in heat shock protein 90 (HSP90) and selected HSP90-chaperoned proteins. METHODS: At specified times after i.v. administration of 75 mg/kg 17DMAG, SCID mice bearing s.c. MDA-MB-231 human breast xenografts were killed and plasma and tissues were retained. 17DMAG concentrations were determined by HPLC. Raf-1, heat shock protein 70 (HSP70), and HSP90 in tissues were determined by Western blotting. RESULTS: Peak plasma 17DMAG concentration was 15.4+/-1.4 microg/ml. The area under the plasma 17DMAG concentration versus time curve was 1072 microg/ml min, corresponding to a total body clearance of 70 ml/kg/min. Peak 17DMAG concentrations in liver (118.8+/-5.7 microg/g), kidney (122.9+/-10.6 microg/g), heart (81.3+/-8.1 microg/g), and lung (110.6+/-25.4 microg/g) occurred at 5-10 min, while peak concentrations in spleen (70.6+/-9.6 microg/g) and tumor (9.0+/-1.0 microg/g) occurred at 30-45 min. At 48 h, 17DMAG was detectable in tumor but not in any normal tissue. Raf-1 in tumors of 17DMAG treated mice killed at 4, 7, 24 and 48 h was about 20% lower than in tumors from vehicle-treated mice. HSP90 and HSP70 in tumors of 17DMAG-treated animals were significantly lower than in tumors of control animals at 4, 7, and 24 h. Hepatic Raf-1 was decreased by more than 60% at all times after 17DMAG treatment; however, hepatic HSP90 was not affected. HSP70 was undetectable in livers of vehicle-treated mice or mice killed at 2 or 4 h after 17DMAG treatment, but was detected in livers at 7, 24 and 48 h. 17DMAG did not affect renal Raf-1. In contrast, renal HSP70 and HSP90 were decreased by more than 50% at 2 and 4 h after 17DMAG treatment. Renal HSP70 increased approximately twofold above that in kidneys from vehicle-treated control mice at 7 and 24 h, while HSP90 relative protein concentration was no different from that in controls. CONCLUSIONS: Plasma pharmacokinetics of 17DMAG in tumor-bearing mice were similar to those previously reported in nontumor-bearing mice. 17DMAG was distributed widely to tissues but was retained for longer in tumors than normal tissues. Raf-1, HSP90, and HSP70 were altered to different degrees in tumors, livers, and kidneys of 17DMAG treated animals. These data illustrate the complex nature of the biological responses to 17DMAG. PMID- 15338193 TI - P-glycoprotein plays a role in the oral absorption of BMS-387032, a potent cyclin dependent kinase 2 inhibitor, in rats. AB - PURPOSE: BMS-387032, a novel cyclin-dependent kinase 2 inhibitor, is currently in phase I clinical trials for anticancer therapy. The oral bioavailability of BMS 387032 has been found to be about 31% in rats. Absorption and first-pass metabolism were evaluated as possible reasons for the incomplete oral bioavailability in rats. METHODS: Male Sprague-Dawley rats were given single doses of BMS-387032 intraarterially (9.1 mg/kg), orally (9.1 mg/kg), or intraportally (10 mg/kg). The routes of excretion of BMS-387032 after intravenous dosing were investigated in bile-duct-cannulated rats. The rate of metabolism of BMS-387032 was investigated in liver microsomes. The permeability of BMS-387032 was evaluated using Caco-2 cells, an in vitro model of the intestinal epithelium. To determine if BMS-387032 was a P-glycoprotein substrate, brain uptake studies were conducted in P-glycoprotein knockout versus wildtype mice. RESULTS: The exposure in rats after an intraportal dose was similar to that after an intraarterial dose, indicating that absorption may play a greater role than liver first-pass metabolism in the low oral bioavailability seen in rats. After an intravenous dose, the percent of dose excreted unchanged in the urine and bile over a 9-h period was 28% and 11%, respectively. In vitro studies in rat liver microsomes showed low rates of metabolism of BMS-387032. The Caco-2 cell permeability of BMS-387032 was <15 nm/s in the apical to basolateral direction, and 161 nm/s in the basolateral to apical direction, indicating that it may be a substrate for an intestinal efflux transporter. A P-glycoprotein binding assay showed that BMS-387032 might be a P-glycoprotein modulator. Brain penetration studies in mice showed brain levels of BMS-387032 about 3.5-fold higher in P glycoprotein knockout mice than in wildtype mice, providing evidence of BMS 387032 being a P-glycoprotein substrate. CONCLUSIONS: Poor absorption may be playing a greater role than extensive first-pass metabolism in the incomplete oral bioavailability of BMS-387032 seen in rats. The efflux transporter, P glycoprotein, may be responsible for limiting absorption, as BMS-387032 appears to be a substrate of P-glycoprotein. PMID- 15338194 TI - Delayed hepatitis B virus reactivation after cessation of preemptive lamivudine in lymphoma patients treated with rituximab plus CHOP. AB - Preemptive lamivudine in lymphoma patients undergoing intensive chemotherapy can effectively prevent chemotherapy-related HBV reactivation. Nevertheless, the safety profile after withdrawal of lamivudine and the impact of rituximab containing chemotherapy on HBV reactivation has not been defined. To illustrate the necessity of prolonged surveillance after cessation of preemptive lamivudine in lymphoma patients treated with rituximab and chemotherapy, four patients with B-cell NHL carrying HBV received rituximab plus CHOP. Preemptive lamivudine therapy was administered 1 week before chemotherapy until 4 weeks after completion of chemotherapy. Serial serum alanine aminotransferase (ALT), total bilirubin, and HBV-DNA levels were prospectively monitored in three patients. The fourth patient was closely monitored for ALT. The HBV DNA was checked after development of clinical overt hepatitis. The peripheral blood CD20+ B-lymphocyte counts were analyzed periodically in two patients. All of the three patients studied prospectively had virological relapses with surgence of HBV DNA 6-8 months after completion of rituximab-plus-CHOP (R+CHOP) therapy. Two of the three patients had biochemical relapses and one of them developed severe hepatitis. Sequencing for HBV polymerase gene in these patients failed to show evident emergence of lamivudine-resistant mutations. The fourth patient developed a hepatitis flare-up 6 months after completion of chemotherapy. The CD2+ lymphocytes were totally depleted when HBV DNA started to increase. Delayed HBV reactivation can occur in lymphoma patients receiving R+CHOP after withdrawal of preemptive lamivudine. More protracted lamivudine therapy may be an alternative to close monitoring following chemotherapy, and further studies are needed to define optimal duration of lamivudine therapy. PMID- 15338195 TI - Chronic idiopathic neutropenia preceding polymyalgia rheumatica and acute myeloid leukemia. PMID- 15338196 TI - Altered erythrocyte membrane characteristics during anemia in childhood acute lymphoblastic leukemia. AB - Anemia is a prominent feature in children with acute lymphoblastic leukemia (ALL). To investigate the erythrocyte features during anemia in these patients, we studied the altered characters of these cells and oxidative stress imposed in their serum. This investigation reveals that erythrocytes from ALL patients show (1) increased membrane fluidity detected by fluorescence anisotropy studies, increased osmotic fragility detected by hemolysis of erythrocytes in different saline concentrations, and increased hydrophobicity as measured by binding with 8 anilino-1-naphthalenesulfonic acid, (2) enhanced (approximately threefold) glycosylation and sialylation, monitored by digoxigenin enzyme assay, and (3) expression of disease-specific 210, 105, 83, 54, and 28 kDa 9-O-acetyl sialoglycoconjugates (9-O-AcSGs) demonstrated by Western blot analysis and fluorescence-activated cell sorter (FACS) analysis studies using Achatinin-H with specificity towards 9-O-AcSAalpha2-6GalNAc as the analytical probe. (4) In addition, induced oxidative stress was observed in the sera of these children as indicated by increased nitric oxide (approximately fourfold) and thiobarbituric acid (TBA) reactive species (twofold) as detected by Griess reaction and TBA assay, respectively. For all the experiments, erythrocytes from normal individuals served as controls. Thus, the altered membrane characteristics together with their exposure to induced oxidative stress in serum are found to be a few features restricted to diseased erythrocytes. Taken together, our results are suggestive of their interplay in the contribution to the observed anemia in these patients, which may be exploited for better management of the disease. PMID- 15338197 TI - Hemodynamic assessment of splenomegaly in beta-thalassemia patients undergoing splenectomy. AB - Splenomegaly is a common finding in beta-thalassemia; however, its hemodynamic features and its potential correlations with high output state and hepatic disorders, both also frequent in thalassemia, have not yet been assessed in these patients. Eight beta-thalassemia patients with the indication for splenectomy and no symptoms or signs of heart disease, aged 25.6+/-5.5 years, were studied. Preoperative assessment included hematological profile, liver biology, hepatitis virus serology, and echocardiography. During splenectomy, splenic artery blood flow and splenic vein pressure were directly measured and liver biopsies were taken. Preoperative echocardiographic data were compared with those of 34 healthy controls. The preoperative cardiac index was significantly elevated in patients (4.8+/-1.3 vs 3.4+/-1.1 l/min per m2 in controls, p<0.001). Splenic blood flow, although increased, was not particularly high, being 285+/-56 ml/min or 0.13+/ 0.04 ml/min per g of splenic mass, representing 4.1+/-0.9% of total cardiac output (CO). Splenic vein pressure was considerably elevated (29.7+/-5.5 cmH2O). Hepatic fibrosis, iron deposition, and extramedullary foci were found in all eight biopsies. Serology was positive in five of eight cases. beta-thalassemia patients with extensive splenomegaly requiring splenectomy are characterized by high output state, increased splenic blood flow, which probably makes a limited contribution to CO elevation, and portal hypertension, manifest by increased splenic vein pressure and hepatic histopathological abnormalities. PMID- 15338198 TI - Prognostic value of urinary pyridinium crosslinks and their derivatives in multiple myeloma. AB - Bone disease is a common feature of multiple myeloma (MM). The goal of this study was to assess the prognostic significance of urinary markers of bone metabolism in MM. Urinary levels of total pyridinoline (T-Pyd), deoxypyridinoline (T-Dpd), crosslinked N-telopeptide (Ntx), C-telopeptide (Ctx) of type I collagen and immunologic free deoxypyridinoline (f-Dpd) were assessed in 82 consecutive, previously untreated MM patients (aged 65-75 years) diagnosed between June 1995 and December 1998. A correlation between disease stage according to the Durie Salmon classification and T-Pyd (p=0.034) and T-Dpd (p=0.007) was observed, while T-Pyd (p=0.015) and to a lesser extent f-Dpd (p=0.081) were correlated to bone involvement measured by plain X-ray. None were correlated to M-component or magnetic resonance imaging (MRI). In univariate analysis high T-Pyd (p=0.007), T Dpd (p=0.027), f-Dpd (p<0.001), and Ctx (p=0.011) were associated with shorter overall survival, whereas only f-Dpd (p=0.0025) was associated with a shorter disease-free survival. In multivariate analysis, C-reactive protein and f-Dpd were independent prognostic factors for overall survival. This retrospective analysis defined new independent prognostic indicators of survival in patients with newly diagnosed myeloma. Indeed, urinary markers of bone resorption can easily be measured at diagnosis and have independent prognostic significance to refine the prognosis of MM patients. PMID- 15338199 TI - Is the antiplatelet effect of aspirin affected by systemic inflammation? PMID- 15338200 TI - Limb salvage in aggressive and malignant tumours of the fibula. AB - We treated 25 patients with aggressive and malignant fibular tumours between April 1989 and May 2001. There were 11 osteosarcomas, seven Ewing's sarcoma and five chondrosarcomas. The tumours predominantly involved the upper one third; 16 were of Stage IIB. Neo-adjuvant and adjuvant chemotherapy were given to all patients with osteosarcoma and Ewing's sarcoma. In 20 patients, surgical margins were wide and in three radical. Mean follow-up was 71 (26-168) months. Local recurrences were met with in three patients requiring amputation. Three patients died of disease and one was lost to follow-up. Functional outcome was excellent in 12 patients and good in seven. The 5- and 10-year survival rates of patient and limb are reported based on Kaplan-Meier survival analysis. PMID- 15338201 TI - Microbiology and management of myositis. AB - This review summarizes the microbiology, management, and prevention of myositis. Muscular infections frequently occur in areas of the body that have been compromised or injured by a foreign body, trauma, ischemia, injection of illicit drug, malignancy, or surgery. These infections can develop very rapidly to life threatening systemic illness. The predominant pathogens are Staphylococcus aureus, Group A streptococci (GAS), gram-negative aerobic and facultative bacilli, and the indigenous aerobic and anaerobic cutaneous and mucous membranes local microflora. Pyogenic myositis can be classified into either GAS necrotizing myositis, clostridial myonecrosis (gas gangrene), or nonclostridial (crepitant) myositis. Intensive surgical and medical therapy that includes the administration of intravenous fluids and antimicrobial therapy is an essential element in management of muscle infection. PMID- 15338203 TI - Proprioception and performance after anterior cruciate ligament rupture. AB - The aim of this study was to investigate the characteristics of proprioception in patients with an anaterior-cruciate-ligament (ACL)-injured knee and to determine whether there is a correlation between proprioception and performance. We studied 32 patients with unilateral isolated ACL ruptures. Proprioception of the knee was evaluated by examining the joint position sense. Functional performance was evaluated with the one-leg hop (OLH) and one-leg vertical jump (OLV) tests. The mean error angle of the joint position sense was 3.6+/-1.5 degrees on the intact side and 5.2+/-1.9 degrees on the injured side. The joint position sense was thus clearly reduced on the injured side (p<0.05). The distance of jumping in the OLH test and the height of jumping in the OLV test was also clearly reduced on the injured side compared with that on the intact side (p<0.01) both with and without visual control. Moreover, we found a significant correlation between proprioception and performance in the ACL-injured knees, and this correlation was more distinct with visual deprivation. In conclusion, decreased proprioception in patients with ACL deficiency reduced their functional ability. PMID- 15338205 TI - One-year follow-up of orthopaedic implants in HIV-positive patients. AB - We followed prospectively 38 orthopaedic implants in 36 HIV-positive patients. X rays and clinical examination were used to assess union, and observation was made for early and late wound sepsis for 12 months from the time of surgery. Two patients died of causes unrelated to the implantation, two patients had implants removed for reasons other than infection and eight cases were lost to follow-up. Of the 26 cases that were reviewed at 1 year, no late sepsis was identified. All of the fractures, non-unions, osteotomies and arthrodeses united. The literature indicates that late sepsis following arthroplasty occurs more frequently in haemophiliacs who are HIV positive than their HIV-negative counterparts. It is still not certain whether or not such a risk also applies to HIV-positive patients who are not haemophiliacs and have undergone internal fixation of fractures or non-unions. This study increases the confidence that fixation in immune-compromised patients with intact skin is safe, at least for the time period that the implant is required. Further studies are required to know whether or not fixation implants should be removed. PMID- 15338202 TI - Valgus impacted proximal humeral fractures and their blood supply after transosseous suturing. AB - We treated 16 patients (11 women and five men, average age 45 years), all having four-part valgus impacted fractures of the proximal humerus, with transosseous suturing. All had preoperative angiography performed 6-12 h after admission. The average impaction angle was 43 degrees , and the mean lateral displacement of the humeral head was 1.4 mm. Postoperative angiography was performed 8-10 weeks after the operation followed by digital image processing using the segmentation technique. No statistically important reduction in the length and area of large (>0.5 mm) vessels was seen. Union was confirmed by the reduction in the length and area of small vessels (<0.5 mm). At a mean follow-up of 40 months, avascular necrosis was only found in one patient. The average Constant-Murley score was 87 (67-100) points, whereas the functional score in comparison with the unaffected shoulder was 94% (89-100%). Despite the small number of patients, transosseous fixation seems to preserve the remaining blood supply of the humeral head. PMID- 15338206 TI - Effective immunotherapy against cancer: a question of overcoming immune suppression and immune escape? AB - During the last decade, the breakthroughs in understanding of the molecular mechanisms responsible for immune activation and the advent of recombinant DNA technologies have changed the view on immunotherapy from "a dream scenario" to becoming a clinical reality. It is now clear that both cellular immunity comprising T and NK cells, as well as strategies based on antibodies, can provide strong antitumoral effects, and evidence is emerging that these strategies may also cure patients with previously incurable cancers. However, there are still a number of issues that remain unresolved. Progress in immunotherapy against cancer requires a combination of new, improved clinical protocols and strategies for overcoming mechanisms of immune escape and tumor-induced immune suppression. This review discusses some of the salient issues that still need to be resolved, focusing on the role of oxidative stress and the use of antioxidants to alleviate the immune hyporesponsiveness induced by reactive oxygen species (ROS). PMID- 15338208 TI - Primary hyperparathyroidism: is there a role for imaging? (Against). PMID- 15338207 TI - Primary hyperparathyroidism: is there a role for imaging? (Pro). PMID- 15338209 TI - Fever of unknown origin: prospective comparison of diagnostic value of (18)F-FDG PET and (111)In-granulocyte scintigraphy. PMID- 15338212 TI - Forearm interosseous membrane imaging and anatomy. AB - OBJECTIVE: To determine the regional thickness variation of the interosseous membrane (IOM) along the forearm and validate magnetic resonance imaging of the IOM with laser micrometry. DESIGN AND PATIENTS: Axial thickness measurements of 12 cadaver forearms were obtained using magnetic resonance imaging (MRI) at radial, central, and ulnar locations. The specimens were dissected, and IOM thickness measured using a laser micrometer. MRI and laser measurements of the main and oblique IOM bundles were compared. An axial thickness profile was plotted versus forearm length, and radial, central, and ulnar positions were compared. RESULTS: The main bundle thickness was 2.18+/-0.20 mm using laser micrometry, which was not significantly different from MRI measurements (1.86+/ 0.25 mm, p=0.11, power = 0.84). The dorsal oblique bundle thickness was not significantly different between measurement methods (2.93+/-0.77 mm and 3.30+/ 1.64 mm using laser micrometry and MRI respectively, p=0.75, power = 0.04). Both methods demonstrated a progressive increase in thickness proximally within the forearm. MRI measurements demonstrated a significantly greater thickness increase in the radial location compared to the central location (slope = 2.26 and 1.05, r(2)=0.31 and 0.12 respectively, p<0.05). The ulnar slope was not significantly different from zero ( r(2)=0.02, p>0.05). CONCLUSION: Our findings describe the varying IOM anatomy using MRI, and determined the location of the clinically important IOM fiber bundles. This study confirms the accuracy of MR imaging of the IOM by comparison with a laser micrometer, and demonstrates the thickness variation along the forearm. This information may be used to identify changes in IOM anatomy with both acute IOM injury and chronic fiber attenuation. PMID- 15338213 TI - Multiple vertebral fluid-fluid levels. AB - We present a case of multiple vertebral metastases, with multiple fluid-fluid levels, from a moderately to poorly differentiated carcinoma of unknown origin. We suggest that fluid-fluid levels in multiple vertebral lesions are highly suggestive of bone metastases. PMID- 15338214 TI - Non-contrast MR imaging of the glenohumeral joint. Part I. Normal anatomy. AB - MR imaging of the shoulder without contrast is frequently used for evaluation of glenohumeral instability in spite of the popularity of MR arthrography. With proper imaging technique, familiarity with normal anatomy and variants as well as knowledge of the expected pathologic findings high diagnostic accuracy may be achieved. PMID- 15338215 TI - Balance control during an arm raising movement in bipedal stance: which biomechanical factor is controlled? AB - In order to obtain new insight into the control of balance during arm raising movements in bipedal stance, we performed a biomechanical analysis of kinematics and dynamical aspects of arm raising movements by combining experimental work, large-scale models of the body, and techniques simulating human behavior. A comparison between experimental and simulated joint kinematics showed that the minimum torque change model yielded realistic trajectories. We then performed an analysis based on computer simulations. Since keeping the center of pressure (CoP) and the projection of the center of mass (CoM) inside the support area is essential for equilibrium, we modeled an arm raising movement where displacement of one or the other variable is limited. For this optimization model, the effects of adding equilibrium constraints on movement trajectories were investigated. The results show that: (a) the choice of the regulated variable influences the strategy adopted by the system and (b) the system was not able to regulate the CoM for very fast movements without compromising its balance. Consequently, we suggest that the system is able to maintain balance while raising the arm by only controlling the CoP. This may be done mainly by using hip mechanisms and controlling net ankle torque. PMID- 15338216 TI - Modeling interactions between photic and nonphotic entrainment mechanisms in transmeridian flights. AB - In transmeridian flights, photic and nonphotic entrainment mechanisms are expected to interact dynamically in the human circadian system. In order to simulate the reentrainment process of the circadian rhythms, the photic entrainment mechanism was introduced to our previous model, which consisted of three coupled oscillators. Regardless of flight direction, a large time difference beyond 10 h tended to induce the antidromic reentrainment. The partition between the oscillators resulted for the eastward flight over a 10-h or longer time difference and the westward over 6 h or longer. The simulated reentrainment processes almost coincided with empirical knowledge. Simulated effects of physical exercise showed that some antidromic reentrainments were switched to the orthodromic ones for the eastward flight and most of the partitions between the oscillators were prevented in the westward flight. These results are due to an augmentation of the entrainment pressure of the rest activity cycle on the oscillators. The mechanisms underlying these various reentrainment patterns were explained based on the photic response, the interactions between the oscillators, and their adaptive modification. The simulation results suggest that an appropriate selection of departure time and physical exercise could ease the jet lag caused by transmeridian flight. PMID- 15338217 TI - Knee extensor and plantar flexor muscle size and function following 90 days of bed rest with or without resistance exercise. AB - Skeletal muscle atrophy and strength loss induced by short-term simulated spaceflight are offset or attenuated by resistance exercise (RE). This study compared the effects of plantar flexor and knee extensor RE on muscle size and function in 17 healthy men (aged 26-41years) subjected to 90 days 6 degrees head down-tilt bed rest with (BRE; n = 8) or without (BR; n = 9) RE. The RE program consisted of coupled maximal concentric and eccentric actions in the supine squat (4 sets of 7 repetitions) and calf press (4 x 14) every third day employing a gravity-independent flywheel ergometer (FW). Prior to, and following bed rest, muscle volume was assessed using magnetic resonance imaging. Similarly, muscle strength and power and surface electromyographic (EMG) activity were determined during maximal actions using FW or isokinetic dynamometry. In BR, knee extensor and plantar flexor muscle volume decreased (P < 0.05) 18% and 29%, respectively. Torque or force and power decreased (P < 0.05) 31 60% (knee extension) and 37-56% (plantar flexion) while knee extensor and plantar flexor EMG activity decreased 31-38% and 28-35%, respectively following BR. Muscle atrophy in BRE was prevented (P > 0.05; knee extensors) or attenuated (-15%; plantar flexors). BRE maintained task-specific force, power and EMG activity. The decrease in non-task-specific torque was less (P < 0.05) than in BR. The present data imply that the triceps surae and quadriceps muscles show different responsiveness to long-term bed rest with or without resistance exercise. The results also suggest that designing in flight resistance exercise protocols for space travellers is complex and must extend beyond preserving muscle only. PMID- 15338218 TI - The role of exercising muscle length in the protective adaptation to a single bout of eccentric exercise. AB - The purpose of this study was to determine if the protective adaptation to a single bout of eccentric exercise (repeated bout effect) is dependent on the muscle length at which the eccentric contractions are performed. Ten subjects (six men, four women) performed two bouts of 120 isokinetic eccentric contractions separated by 2 weeks (target intensity was 90% of maximum isometric strength at 70 degrees). In the initial bout one limb exercised from 30 degrees to 70 degrees of knee flexion (short initial bout; SIB) and the contralateral limb exercised from 70 degrees to 110 degrees (long initial bout; LIB). For the repeated bout 2 weeks later, the limb that initially exercised at a short length now exercised at a long length (long repeated bout; LRB) and the limb that initially exercised at a long length now exercised at a short length (short repeated bout; SRB). Isometric strength and pain (scale 0-10) were assessed immediately post exercise and on the next 3 days. Strength loss and pain were greater following LIB versus SIB (strength loss P < 0.01; pain P < 0.001) and following LRB versus SRB (strength loss P < 0.01; pain P < 0.001). Strength loss and pain were not different between LIB and LRB. Pain was significantly greater following SIB compared with SRB (P < 0.05). Strength loss was not different between SIB and SRB. These results confirm that the symptoms of muscle damage are highly dependent on exercising muscle length and also demonstrate that the repeated bout effect is dependent on muscle length. Performing an initial bout of eccentric exercise at a shortened muscle length did not protect against strength loss and pain following a repeated bout at a longer muscle length. Data are given as mean (SE) unless otherwise stated. PMID- 15338219 TI - Convective heat transfer area of the human body. AB - In order to clarify the heat transfer area involved in convective heat exchange for the human body, the total body surface area of six healthy subjects was measured, and the non-convective heat transfer area and floor and chair contact areas for the following nine common body positions were measured: standing, sitting on a chair, sitting in the seiza position, sitting cross-legged, sitting sideways, sitting with both knees erect, sitting with a leg out, and the lateral and supine positions. The main non-convective heat transfer areas were: the armpits (contact between the upper arm and trunk regions), contact between the two legs, contacts between the fingers and toes, and contact between the hands and the body surface. Also, when sitting on the floor with some degree of leg contact (sitting in the seiza position, cross-legged, or sideways), there was a large non-convective heat transfer area on the thighs and legs. Even when standing or sitting in a chair, about 6-8% of the body surface did not transfer heat by convection. The results showed that the effective thermal convective area factor for the naked whole body in the standing position was 0.942. While sitting in a chair this factor was 0.860, while sitting in a chair but excluding the chair contact area it was 0.918, when sitting in the seiza position 0.818, when sitting cross-legged 0.843, in the sideways sitting position 0.855, when sitting with both knees erect 0.887, in the leg-out sitting position 0.906, while in the lateral position it was 0.877 and the supine position 0.844. For all body positions, the effective thermal convective area factor was greater than the effective thermal radiation area factor, but smaller than the total body surface area. PMID- 15338220 TI - Heart rate variability and its relation to prefrontal cognitive function: the effects of training and detraining. AB - The aim of the present study was to investigate the relationship between physical fitness, heart rate variability (HRV) and cognitive function in 37 male sailors from the Royal Norwegian Navy. All subjects participated in an 8-week training program, after which the subjects completed the initial cognitive testing (pretest). The subjects were assigned into a detrained group (DG) and a trained group (TG) based on their application for further duty. The DG withdrew from the training program for 4 weeks after which all subjects then completed the cognitive testing again (post-test). Physical fitness, measured as maximum oxygen consumption (VO2(max)), resting HRV, and cognitive function, measured using a continuous performance task (CPT) and a working memory test (WMT), were recorded during the pre-test and the post-test, and the data presented as the means and standard deviations. The results showed no between-group differences in VO2(max) or HRV at the pre-test. The DG showed a significant decrease in VO2(max) from the pre- to the post-test and a lower resting HRV than the TG on the post-test. Whereas there were no between-group differences on the CPT or WMT at the pre test, the TG had faster reaction times and more true positive responses on tests of executive function at the post-test compared to the pre-test. The DG showed faster reaction times on non-executive tasks at the post-test compared to the pre test. The results are discussed within a neurovisceral integration framework linking parasympathetic outflow to the heart to prefrontal neural functions. PMID- 15338221 TI - Infrared thermography for examination of skin temperature in the dorsal hand of office workers. AB - Reduced blood flow may contribute to the pathophysiology of upper extremity musculoskeletal disorders (UEMSD), such as tendinitis and carpal tunnel syndrome. The study objective was to characterize potential differences in cutaneous temperature, among three groups of office workers assessed by dynamic thermography following a 9-min typing challenge: those with UEMSD, with ( n=6) or without ( n=10) cold hands exacerbated by keyboard use, and control subjects ( n=12). Temperature images of the metacarpal region of the dorsal hand were obtained 1 min before typing, and during three 2-min sample periods [0-2 min (early), 3-5 min (middle), and 8-10 min (late)] after typing. Mean temperature increased from baseline levels immediately after typing by a similar magnitude, 0.7 (0.3) degrees C in controls and 0.6 (0.2) degrees C in UEMSD cases without cold hands, but only by 0.1 (0.3) degrees C in those with cold hands. Using paired t-tests for within group comparisons of mean dorsal temperature between successive imaging periods, three patterns of temperature change were apparent during 10 min following typing. Controls further increased mean temperature by 0.1 degrees C ( t-test, P=0.001) at 3-5 min post-typing before a late temperature decline of -0.3 degrees C ( t-test, P=0.04), while cases without cold hands showed no change from initial post-typing mean temperature rise during middle or late periods. In contrast, subjects with keyboard-induced cold hands had no change from initial post-typing temperature until a decrease at the late period of -0.3 degrees C ( t-test, P=0.06). Infrared thermography appears to distinguish between the three groups of subjects, with keyboard-induced cold hand symptoms presumably due, at least partially, to reduced blood flow. PMID- 15338222 TI - Risk of health complaints and disabilities among Dutch firefighters. AB - OBJECTIVES: The purpose of the present study was threefold: (1) to compare the work demands on firefighters (FFs) and office workers (OWs), (2) to compare the prevalence of health complaints and disabilities in the work situation in these two groups, and (3) to explore the effect of work demands on the risk of health complaints. METHODS: Self-reported information was gathered from 1,624 FFs (55% response) and 630 OWs (80% response), at the same fire departments in different regions of the Netherlands, on work demands ('sitting', and biomechanically and energetically demanding activities and 24-h shifts), health complaints and disabilities. First, we compared the work demands and prevalence rates of health complaints and related disabilities in the two groups, then we explored the risk of health complaints in workers with high and low exposure to work demands. RESULTS: Compared with office workers, FFs reported: (1) less exposure to 'sitting' and more to biomechanically and energetically demanding activities, (2) more knee (OWs 14% vs FFs 20%) and ankle (3% vs 10%) complaints and disabilities resulting from back complaints (30% vs 47%), and (3) less hypertension (7% vs 5%), stomach (13% vs 7%), heart (6% vs 2%), neck (26% vs 16%), shoulder (16% vs 14%) and arm (14% vs 6%) complaints. A higher risk of subjective fatigue was found in workers highly exposed to 'energetically demanding activities', and of neck, shoulder and arm complaints in workers highly exposed to 'sitting'. CONCLUSIONS: Firefighters reported higher physical demands (with the exception of 'sitting') than office workers did. The prevalence rate of certain complaints or disabilities among FFs was higher (knee and ankle complaints and disabilities related to back complaints) or lower (hypertension, stomach, heart, neck, shoulder and arm complaints) than among OWs. The results suggest that exposure to highly biomechanically demanding activities might cause an increased risk of knee and ankle complaints and that exposure to highly energetically demanding activities might increase the risk of subjective fatigue. PMID- 15338223 TI - Relation between children's height and outdoor air pollution from coal-burning sources in the British 1946 birth cohort. AB - OBJECTIVE: Air pollution is associated with a number of health outcomes in childhood. In this study, we investigated whether air pollution is related to children's height. METHODS: The 1946 British birth cohort study recruited 5,362 children born in 1 week in March 1946. Height was measured when the children were aged 2, 4, 6, 7, 11 and 15 years. Data on socio-economic conditions and other characteristics were obtained in interviews. Areas of children's residence were categorised into four groups of air pollution on the basis of published coal consumption data. RESULTS: After controlling for socio-economic factors, we found that air pollution was associated with children's height at several ages. The association, adjusted for socio-economic factors, was strongest at the age of 7 years, when children in the most polluted areas were 1.2 (95% confidence interval 0.5-1.8) cm shorter than those in the least polluted areas. After the age of 7 years the effect of air pollution diminished and disappeared by the age of 15 years. Further adjustment for birth weight and respiratory illness in childhood did not change this pattern. CONCLUSIONS: Children's height was inversely associated with air pollution, but the magnitude of the effect depended on age. However, the biological mechanisms linking children's growth with air pollution are not evident, and it remains to be confirmed whether the relationship is genuine and causal. PMID- 15338224 TI - Ecological study on residences in the vicinity of AM radio broadcasting towers and cancer death: preliminary observations in Korea. AB - OBJECTIVES: Public health concern about the health effects of radio-frequency electromagnetic fields (RF-EMFs) has increased with the increase in public exposure. This study was to evaluate some health effect of RF exposure by the AM radio broadcasting towers in Korea. METHODS: We calculated cancer mortality rates using Korean death certificates over the period of 1994-1995 and population census data in ten RF-exposed areas, defined as regions that included AM radio broadcasting towers of over 100 kW, and in control areas, defined as regions without a radio broadcasting tower inside and at least 2 km away from the towers. RESULTS: All cancers-mortality was significantly higher in the exposed areas [direct standardized mortality rate ratio (MRR) = 1.29, 95%CI = 1.12-1.49]. When grouped by each exposed area and by electrical power, MRRs for two sites of 100 kW, one site of 250 kW and one site of 500 kW, for all subjects, and for one site of 100 kW and two sites of 250 kW, for male subjects, showed statistically significant increases without increasing trends according to the groups of electric power. Leukemia mortality was higher in exposed areas (MRR = 1.70, 95% CI = 0.84-3.45), especially among young adults aged under 30 years (0-14 years age group, MRR = 2.29, 95% CI = 1.05-5.98; 15-29 age group, MRR = 2.44, 95% CI = 1.07-5.24). CONCLUSIONS: We observed higher mortality rates for all cancers and leukemia in some age groups in the area near the AM radio broadcasting towers. Although these findings do not prove a causal link between cancer and RF exposure from AM radio broadcasting towers, it does suggest that further analytical studies on this topic are needed in Korea. PMID- 15338225 TI - Psychological well-being among hospital personnel: the role of family demands and psychosocial work environment. AB - BACKGROUND: This study investigates the effect of gender role and the psychosocial work environment on the psychological well-being of hospital staff in two general hospitals in the province of Valencia (Spain). METHOD: A cross sectional survey was carried out among 313 workers by means of a self-answered questionnaire. The outcome variable (psychological well-being) was evaluated with four dimensions of the "SF-36 Health Survey" (mental health, vitality, limitations in the emotional role and limitations in the social function). The explanatory variables were: characteristics related to gender role, professional characteristics and the psychosocial working environment evaluated according to Karasek and Johnson's demand-control-support model. The adjusted odds ratios (ORs) and their 95% confidence intervals were calculated by logistical regression. RESULTS: Those who have very good marital relationship have less risk of presenting bad mental health, OR 0.43 (0.24-0.78), and limitation in the social function, OR 0.43 (0.24-0.77), and in the emotional role, OR 0.35 (0.16 0.74). Those who dedicate more than 30 h a week to domestic chores have a higher risk of limitation of social function, OR 2.48 (1.16-5.31). Those exposed to high psychological demands present a higher probability of having bad mental health, OR 1.77 (1.04-3.00). Those exposed to low job social support have a higher risk of bad mental health, OR 1.86 (1.09-3.19), low vitality, OR 2.21 (1.30-3.77), and limitation in the social function, OR 1.88 (1.10-3.22). CONCLUSION: Gender role and psychosocial work environment have a negative influence on the psychological well-being of hospital staff. PMID- 15338226 TI - Nitric oxide insufficiency and atherothrombosis. AB - Nitric oxide (NO) is a structurally simple compound that participates in a wide range of biological reactions to maintain normal endothelial function and an antithrombotic intravascular milieu. Among its principal effects are the regulation of vascular tone, vascular smooth muscle cell proliferation, endothelial-leukocyte interactions, and the antiplatelet effects of the endothelium. Impaired NO bioavailability represents the central feature of endothelial dysfunction, the earliest stage in the atherosclerotic process, and also contributes to the pathogenesis of acute vascular syndromes by predisposing to intravascular thrombosis. The causes of NO insufficiency can be grouped into two fundamental mechanisms: inadequate synthesis and increased inactivation of NO. Polymorphisms in the endothelial NO synthase gene and decreased substrate or cofactor availability for this enzyme are the main mechanisms that compromise the synthesis of NO. Inactivation of NO occurs mainly through its interaction with reactive oxygen species and can be favored by a deficiency of antioxidant enzymes such as glutathione peroxidase. In this review, we present an overview of NO synthesis and biological chemistry, discuss the mechanisms of action of NO in regulating endothelial and platelet function, and explore the causes of NO insufficiency, as well as the evidence linking these causes to the pathophysiology of endothelial dysfunction and atherothrombosis. PMID- 15338227 TI - Immunocytochemical properties of stellate ganglion neurons during early postnatal development. AB - Neurotransmitter features in sympathetic neurons are subject to change during development. To better understand the neuroplasticity of sympathetic neurons during early postnatal ontogenesis, this study was set up to immunocytochemically investigate the development of the catecholaminergic, cholinergic, and peptidergic phenotypes in the stellate ganglion of mice and rats. The present study was performed on Wistar rats and Swiss mice of different ages (newborn, 10 day-old, 20-day-old, 30-day-old, and 60-day-old). To this end, double labeling for tyrosine hydroxylase (TH), choline acetyltransferase (ChAT), vasoactive intestinal (poly)peptide (VIP), neuropeptide Y (NPY), galanin (GAL), and somatostatin (SOM) was applied. The results obtained indicate that the majority of the neurons in the stellate ganglion of both species were TH-positive from birth onward and that a large part of these neurons also contained NPY. The percentage of neurons containing TH and NPY invariably increased with age up to 60 days postnatally. A smaller portion of the stellate ganglion neurons contained other types of neuropeptides and showed a distinct chronological pattern. The proportion of VIP- and ChAT-positive neurons was maximal in 10-day-old animals and then decreased up to 60 days of age, whereas the number of SOM-positive cells in rats significantly decreased from birth onward. In newborn rats, VIP-, ChAT- and SOM-positive neurons were largely TH-positive, while their proportions decreased in 10-day-old and older rats. Accordingly, the largest part of VIP positive neurons also expressed SOM immunoreactivity at birth, after which the number of neurons containing both peptides diminished. The VIP- and SOM-positive cells did not contain NPY in any of the age groups studied. In rats up to 10 days of life, GAL-immunoreactive (-IR) neurons were scarce, after which their number increased to reach a maximal value in 30-day-old animals and then declined again. The SOM-reactive cells had the smallest size in all rats, while the largest neurons were those containing ChAT. In the mouse stellate ganglion, VIP- and ChAT IR neurons were larger in comparison to NPY- and TH-IR cells. Our study further revealed some species differences: compared to mice the proportion of neurons containing TH and NPY was higher in rats at all ages under study. Furthermore, no GAL-immunostained neurons were found in mice and the number of SOM-positive cells in mice was limited compared to that observed in rats. In conclusion, the development of neurotransmitter composition is complete in rats and mice by their second month of life. At this age, the percentages of immunopositive cells have become similar to those reported in adult animals. PMID- 15338228 TI - Cultivation of human tenocytes in high-density culture. AB - Limited supplies of tendon tissue for use in reconstructive surgery require development of phenotypically stable tenocytes cultivated in vitro. Tenocytes in monolayer culture display an unstable phenotype and tend to dedifferentiate, but those in three-dimensional culture may remain phenotypically and functionally differentiated. In this study we established a three-dimensional high-density culture system for cultivation of human tenocytes for tissue engineering. Human tenocytes were expanded in monolayer culture before transfer to high-density culture. The synthesis of major extracellular matrix proteins and the ultrastructural morphology of the three-dimensional cultures were investigated for up to 2 weeks by electron microscopy, immunohistochemistry, immunoblotting and quantitative, real-time PCR. Differentiated tenocytes were able to survive over a period of 14 days in high-density culture. During the culture period tenocytes exhibited a typical tenocyte morphology embedded in an extensive extracellular matrix containing cross-striated collagen type I fibrils and proteoglycans. Moreover, expression of the tendon-specific marker scleraxis underlined the tenocytic identity of these cells. Taken together, we conclude that the three-dimensional high-density cultures may be useful as a new approach for obtaining differentiated tenocytes for autologous tenocyte transplantation to support tendon and ligament healing and to investigate the effect of tendon affecting agents on tendon in vitro. PMID- 15338230 TI - Localization of HB9 homeodomain protein and characterization of its nuclear localization signal during chick embryonic skin development. AB - We detected HB9 protein during tarsometatarsal scale skin and late feather development. Immunofluorescent analyses with N-terminal 14 amino acids antiserum revealed that HB9 was strongly expressed in epidermal basal cells of the outer scale face in tarsometatarsal scale skin. Specific expression was also detected in dermal cells at the root region of the feather and around the feather follicle. Furthermore, we observed precise distribution of HB9 protein by immunoelectron microscopy. We detected HB9 protein not only in the nucleus, but also in the cytoplasm in tarsometatarsal scale skin. However, in feather skin HB9 protein was found in the nucleus but not in the cytoplasm. Cytoplasmic localization of HB9 protein in tarsometatarsal scale skin was observed especially in the endoplasmic reticulum and the Golgi apparatus. To address the mechanism of nuclear-cytoplasmic translocation, we determined the nuclear localization signal (NLS) sequences by using eukaryotic green fluorescent protein fusion protein in primary keratinocyte culture. Chick HB9 homeoprotein has two types of the NLS sequences in its homeodomain. One of them is a bipartite type as representatively found in Xenopus nucleoplasmin. The other is very similar to hexapeptide NLS sequences identified in pancreatic duodenum homeobox 1 (PDX1). These sequences functioned not only in keratinocytes but also in dermal fibroblasts. They are conserved in Xenopus, mouse, and human HB9 ortholog. These results indicate that HB9 protein might be involved in chick tarsometatarsal scale and feather development and that nuclear import of HB9 protein might be regulated by these NLS sequences in the homeodomain. PMID- 15338232 TI - Low-dose anesthesia for corneal transplantation in mice. AB - BACKGROUND: Despite technical difficulties caused by the small dimensions, scientific and economic considerations stimulated activities in the murine keratoplasty model. Delicate surgery requires stable anesthesia with rapid postoperative waking. METHODS: For corneal transplantation, all animals were intraperitoneally injected with 0.08 ml of a 1:8 mixture of 2% xylazine and 2.5% esketamine hydrochloride. Thus they all received 0.18 mg of xylazine and 1.8 mg of ketamine. In other words, 4.5 mg/kg BW of xylazine and 45 mg/kg BW of ketamine were injected in a mouse weighing 25 g. The inhalation anesthetic isoflurane was used for short interventions like removing lid sutures or in the case of an animal awaking before the end of the operation. A small but effective set of apparatus comprising a glass bottle, a test tube and several interconnected syringes was developed to create a closed system and visualize the amount of isoflurane used in the animal. Air bubbles showed the amount of isoflurane escaping from the opened respiratory mask. RESULTS: The dose reduction of systemic anesthesia requires additional inhalation anesthesia with isoflurane in nearly all cases. The equipment developed is easy to handle and allows systemic anesthesia with a very low dose. CONCLUSION: This murine anesthesia model may be expected to help other surgeons performing corneal transplantations in mice. PMID- 15338229 TI - Reactive oxygen species in vascular biology: implications in hypertension. AB - Reactive oxygen species (ROS), including superoxide (*O2-), hydrogen peroxide (H2O2), and hydroxyl anion (OH-), and reactive nitrogen species, such as nitric oxide (NO) and peroxynitrite (ONOO-), are biologically important O2 derivatives that are increasingly recognized to be important in vascular biology through their oxidation/reduction (redox) potential. All vascular cell types (endothelial cells, vascular smooth muscle cells, and adventitial fibroblasts) produce ROS, primarily via cell membrane-associated NAD(P)H oxidase. Reactive oxygen species regulate vascular function by modulating cell growth, apoptosis/anoikis, migration, inflammation, secretion, and extracellular matrix protein production. An imbalance in redox state where pro-oxidants overwhelm anti-oxidant capacity results in oxidative stress. Oxidative stress and associated oxidative damage are mediators of vascular injury and inflammation in many cardiovascular diseases, including hypertension, hyperlipidemia, and diabetes. Increased generation of ROS has been demonstrated in experimental and human hypertension. Anti-oxidants and agents that interrupt NAD(P)H oxidase-driven *O2- production regress vascular remodeling, improve endothelial function, reduce inflammation, and decrease blood pressure in hypertensive models. This experimental evidence has evoked considerable interest because of the possibilities that therapies targeted against reactive oxygen intermediates, by decreasing generation of ROS and/or by increasing availability of antioxidants, may be useful in minimizing vascular injury and hypertensive end organ damage. The present chapter focuses on the importance of ROS in vascular biology and discusses the role of oxidative stress in vascular damage in hypertension. PMID- 15338233 TI - Autosomal control of the Y-chromosome kl-3 loop of Drosophila melanogaster. AB - The Y chromosome of Drosophila melanogaster carries a limited number of loci necessary for male fertility that possess a series of unconventional features that still hinder a definition of their biological role: they have extremely large sizes; accommodate huge amounts of repetitive DNA; and develop prominent, lampbrush-like loops that bind a number of non-Y-encoded proteins. To obtain insight into the functional role of the loop-forming fertility factors, we characterized four autosomal male-sterile mutations that identify two loci we named loop unfolding protein-1 (lup-1) and loop unfolding protein-2 (lup-2). Biochemical and ultrastructural analysis revealed that neither of them impairs the synthesis of the putative dynein subunit encoded by the ORF localized within the kl-3 fertility factor. However, the stability of four dynein heavy chains is simultaneously affected in each mutant, together with the regular assembly of the axonemal dynein arms that are either absent or strongly reduced. These results indicate that the synthesis of the kl-3-encoded dynein can be uncoupled from the formation of the corresponding loop and suggest that this structure does not simply represent the cytological counterpart of a huge transcription unit, but must be regarded as a complex organelle serving some additional function necessary for male fertility. PMID- 15338234 TI - The mammalian circadian timing system: from gene expression to physiology. AB - Many physiological processes in organisms from bacteria to man are rhythmic, and some of these are controlled by self-sustained oscillators that persist in the absence of external time cues. Circadian clocks are perhaps the best characterized biological oscillators and they exist in virtually all light sensitive organisms. In mammals, they influence nearly all aspects of physiology and behavior, including sleep-wake cycles, cardiovascular activity, endocrinology, body temperature, renal activity, physiology of the gastro intestinal tract, and hepatic metabolism. The master pacemaker is located in the suprachiasmatic nuclei, two small groups of neurons in the ventral part of the hypothalamus. However, most peripheral body cells contain self-sustained circadian oscillators with a molecular makeup similar to that of SCN (suprachiasmatic nucleus) neurons. This organization implies that the SCN must synchronize countless subsidiary oscillators in peripheral tissues, in order to coordinate cyclic physiology. In this review, we will discuss some recent studies on the structure and putative functions of the mammalian circadian timing system, but we will also point out some apparent inconsistencies in the currently publicized model for rhythm generation. PMID- 15338235 TI - Correlation of meiotic events in testis sections and microspreads of mouse spermatocytes relative to the mid-pachytene checkpoint. AB - In mouse, asynaptic meiotic mutants arrest at Testis Epithelial Stage IV. This arrest is 4.5 days after homologous chromosomes begin to synapse and approximately 2.5 days after synapsis is usually completed. To correlate cytological events with meiotic progression in testis and to determine which meiotic events are normally completed by Stage IV, we induced spermatogenic arrest by placing mice on a vitamin A deficient diet. Subsequent injection of retinoic acid and a return to a normal diet resulted in resumption of spermatogenesis with all spermatocytes proceeding through meiosis in a highly synchronous cohort. Between Days 11 and 16 post-injection we prepared one testis for immunocytological and the other for histological evaluation, then used antibodies to SCP3 and either RPA, or MLH1 to follow quantitative changes in synapsis and recombination. RPA was found at sites along the synaptonemal complex as soon as homologs synapsed, and most, but not all, RPA disappeared by Stage IV. MLH1 foci appeared between Stage II and IV and remained through Stage VII, the end point of the study. The data suggest that the earliest the mid-pachytene checkpoint can be activated is Epithelial Stage IV, but that activities monitored by the checkpoint may not be completed by this time. PMID- 15338236 TI - LINE-1 distribution in Afrotheria and Xenarthra: implications for understanding the evolution of LINE-1 in eutherian genomes. AB - Long interspersed nuclear elements (LINEs) comprise about 21% of the human genome (of which L1 is most abundant) and are preferentially accumulated in AT-rich regions, as well as the X and Y chromosomes. Most knowledge of L1 distribution in mammals is restricted to human and mouse. Here we report the first investigation of L1 distribution in the genomes of a wide variety of eutherian mammals, including species in the two basal clades, Afrotheria and Xenarthra. Our results show L1 accumulation on the X of all eutherian mammals, an observation consistent with an ancestral involvement of these elements in the X-inactivation process (the Lyon repeat hypothesis). Surprisingly, conspicuous accumulation of L1 in AT rich regions of the genome was not observed in any species outside of Euarchontoglires (represented by human, mouse and rabbit). Although several features were common to most species investigated, our comprehensive survey shows that the patterns observed in human and mouse are, in many aspects, far from typical for all mammals. We discuss these findings with reference to models that have previously been proposed to explain the AT distribution bias of L1 in human and mouse, and how this relates to the evolution of these elements in other eutherian genomes. PMID- 15338237 TI - Schizosaccharomyces pombe replication protein Cdc45/Sna41 requires Hsk1/Cdc7 and Rad4/Cut5 for chromatin binding. AB - Cdc45 is a conserved protein required for firing of replication origins and processive DNA replication. We used an in situ chromatin-binding assay to determine factors required for fission yeast Cdc45p chromatin binding. Assembly of the pre-replicative complex is essential for Cdc45p chromatin binding, but pre replicative complex assembly occurs independently of Cdc45p. Fission yeast Cdc45p associates with MCM proteins in asynchronously growing cells and cells arrested in S phase by hydroxyurea, but not in cells arrested at the G2/M transition. Both hsk1+ (the fission yeast CDC7 homologue) and rad4+/ cut5+ (the fission yeast DPB11 homologue) are required for Cdc45p chromatin binding. Cdc45p also remains chromatin-bound in mutants that fail to recover from replication arrest. In summary, Cdc45p chromatin binding requires an intact pre-replicative complex as well as signaling from both the Dbf4-dependent kinase and cyclin-dependent kinases. PMID- 15338238 TI - Mismatch repair gene promoter methylation and expression in hydatidiform moles. AB - METHODS: In this study, to investigate the significance of mismatch repair genes (MMR) promoter methylation and expression in the pathogenesis and malignant transformation of hydatidiform moles, we assayed promoter methylation and protein expression of the MMR genes hMLH1 and hMSH2 in gestational trophoblastic diseases (GTDs). DNA was extracted from normal placentas, partial hydatidiform moles, complete hydatidiform moles, and invasive moles, over-digested by methylation sensitive endonuclease Hpa II, and then the promoters were amplificated by polymerase chain reaction. The protein expression was detected by immunohistochemistry. RESULTS: In the normal placentas, neither hMLH1 nor hMSH2 promoter methylation was detected. Expression of hMLH1 and hMSH2 in cytotrophoblasts was strongly positive. In partial hydatidiform moles and complete hydatidiform moles, hMLH1 and hMSH2 promoter methylation rates were significantly higher than that of normal placentas (P = 0.000), and the protein expression in cytotrophoblasts was significantly lower (P = 0.000). In the invasive moles, hMLH1 and hMSH2 promoter methylation was not significantly different compared with the partial hydatidiform moles and complete hydatidiform moles (P > 0.05). Expression of hMLH1 in the invasive moles (54.5%, 6 out of 11) was not significantly different compared with the partial hydatidiform moles and complete hydatidiform moles (P > 0.05). But hMSH2 expression in the invasive moles (36.5%, 4 out of 11) was weaker than that in complete hydatidiform moles (P = 0.044). Promoter methylation and less expression of hMSH2 were correlated in complete hydatidiform moles (P = 0.001) and invasive moles (P = 0.039). CONCLUSIONS: These results indicated that strong expression of hMLH1 and hMSH2 in the cytotrophoblasts of normal placentas may maintain genome stability. Promoter methylation and down-regulation of the expression of hMLH1 and hMSH2 are probably involved in the pathogenesis of hydatidiform moles. PMID- 15338239 TI - Gene expression profiles of three different models of reconstructed human epidermis and classical cultures of keratinocytes using cDNA arrays. AB - The gene expression profiles of three different models of reconstructed human epidermis were analyzed in a comparative study using cDNA array technology. The study also included normal human subconfluent keratinocytes cultured on plastic. Arrays were custom-made and comprised 504 known genes related to cutaneous biology. The gene expression profiles of the three reconstructed epidermis models shared 86% similarity; only 22 of the 504 examined genes showed a different expression level. A comparison of the 3D models with keratinocyte cultures on plastic dishes revealed a set of six genes with a considerably higher expression in the 3D models. These genes were keratin 1, corneodesmosin, filaggrin, loricrin, calmodulin-like skin protein and caspase 14, all related to keratinocyte terminal differentiation. The reported data may contribute to a better understanding and characterization of reconstructed epidermal models and may also serve as established references for investigations related to epidermal differentiation and proliferation. PMID- 15338240 TI - Expression of epidermal N-methyl-D-aspartate receptors (NMDAR1) depends on formation of the granular layer--analysis in diseases with parakeratotic cornification. AB - Ionotrope glutamate receptors of the N-methyl-D-aspartate (NMDA) receptor type are expressed on keratinocytes and influence the intracellular calcium concentration. The importance of NMDA receptors in pathophysiological processes in the skin is, however, still unclear. Epidermal distribution patterns of NMDA receptors were investigated in dermatoses with parakeratotic cornification (psoriasis vulgaris and verrucae vulgares) and compared to the expression of filaggrin. The expression of NMDA receptors (R1 component) in paraffin-embedded normal epidermis (n = 22), psoriasis vulgaris (n = 21) and verrucae vulgares (n = 23) was examined and evaluated by means of digital image analysis. For quantitative characterization of the distribution patterns, a quotient was formed of the expression in the stratum granulosum and stratum basale ("NMDA ratio"). The distribution of NMDAR1 was compared to the immunohistochemical expression of filaggrin. Additionally the expression of filaggrin was investigated in HaCaT cells after treatment with the NMDA receptor antagonist MK-801. NMDA receptors were demonstrated in the epidermis of all preparations. In healthy skin, the highest receptor density was found in the stratum granulosum. This distribution pattern was basically also present in the dermatoses examined. Thus, the occurrence of parakeratosis in psoriasis vulgaris, but not in verrucae vulgares, was characterized by a significant reduction in the NMDA ratio (reduced expression of NMDAR1 in the upper epidermis). The immunohistochemical distribution of filaggrin was similar to that of NMDAR1. In HaCaT cells MK-801 suppressed the expression of filaggrin. NMDA receptors are expressed in human epidermis under physiological conditions especially in the stratum granulosum. Their reduced expression within parakeratotic epidermis in psoriasis vulgaris may be evidence of impaired intracellular calcium influx in this disease. PMID- 15338241 TI - Is screw augmentation necessary in ankle fractures? G. Andreassen et al. "The use of a synthetic bone void filler to augment screws in osteopenic ankle fracture fixation.". PMID- 15338242 TI - Recovery of shoulder joint function after humeral shaft fracture: a comparative study between antegrade intramedullary nailing and plate fixation. AB - INTRODUCTION: Antegrade intramedullary (IM) nailing of humeral shaft fractures is reported to cause shoulder joint impairment. This retrospective study compared shoulder joint symptoms, range of motion (ROM), and isometric strength after antegrade IM nailing and dynamic compression (DC) plating of humeral shaft fractures. MATERIALS AND METHODS: We compared 29 patients with DC plating and 44 with antegrade IM nailing of their humeral shaft fractures. Shoulder pain, L'Insalata and Constant scores, shoulder joint ROM and isometric shoulder strengths were measured after mean follow-up of 6.2 (1-15) years (DC plating) and 5.5 (2-10) years (IM nailing). RESULTS: Patients had nonsignificantly more shoulder pain after IM nailing than after DC plating. Shoulder scores and isometric strength measurements showed no difference between the groups. Flexion was significantly better after DC plating, but none of the other ROM parameters differed between the groups. The shoulder scores and all ROM and strength parameters of the injured side were significantly lower than on the uninjured side in both groups. CONCLUSIONS: Shoulder joint ROM and strength does not recover to normal after humeral shaft fracture. Antegrade IM nailing if performed properly is not responsible for shoulder joint impairment. PMID- 15338243 TI - Displaced supracondylar humeral fractures in children: open reduction vs. closed reduction and pinning. AB - INTRODUCTION: In the literature the best results for pediatric supracondylar humerus fractures have been achieved by closed reduction and wire fixation. However, in these reports the patient group of open reduction and pinning contained the patients who had had previous ineffective closed reduction trials. This retrospective study compared open and closed reduction with pinning, in which the first group of patients was all consecutively treated with open reduction. MATERIALS AND METHODS: The study included 99 children with displaced extension-type supracondylar fractures of humerus who had complete follow-up. Open reduction patients had not had a previous attempted closed reduction. Open reduction and pinning were performed through a posteromedial incision in the first 44 patients and closed reduction and pinning in the subsequent 55 patients. Mean duration surgery was 15 h with open reduction and 17 h with closed reduction. Mean follow up was 35 months with the open reduction and 21 months with closed reduction. Humeral-ulnar angle was compared to the contralateral elbow, clinical flexion deficiency and extension lag, and complications were evaluated. RESULTS: At the latest follow-up the open group had an average of 5.1 degrees valgus change and the closed group 3.6 degrees valgus change in humeral ulnar angle compared to their uninvolved elbow. Average flexion deficiency was 8.61 degrees in the open and 5.25 degrees in the closed group. Average extension lag was 6.23 degrees in the open and 0.6 degrees in the closed group. Functional results were satisfactory in 71% of patients in the open and 93% of those in the closed reduction group. Cosmetic results were satisfactory in 95% of both groups. CONCLUSIONS: Closed reduction and pinning is superior to open reduction and pinning for the treatment of pediatric supracondylar humerus fractures. In the case of technical insufficiencies open reduction and pinning through a posteromedial incision is an alternative treatment for decreasing the surgical time and complications. Complications was not caused in either group by the delayed surgical timing compared to reports in the literature. PMID- 15338244 TI - Pin site care in external fixation sodium chloride or chlorhexidine solution as a cleansing agent. AB - INTRODUCTION: Pin site infection is the most common complication using external fixators. This study investigated the differences in pin site infections, antibiotic use, pain, and complications using sodium chloride and chlorhexidine solution as cleansing agent in patients operated on by the hemicallotasis technique for knee deformities. MATERIALS AND METHODS: The prospective study included 49 consecutive patients: 2 mg/ml chlorhexidine solution was used as cleansing agent in 30 patients (120 pins) and 9 mg/ml sodium chloride in 19 patients (76 pins). We evaluated the status of the pin sites, pain (VAS), uses of antibiotic and analgesic agents, and any complications (infections were graded according to the Checketts-Otterburns classification). Bacterial cultures were performed from each pin site at 1, 6, and 10 weeks and from the pins at removal. RESULTS: Grade 1 infection was found in 14% of the sodium chloride group and in 8.5% of the chlorhexidine group, and grade 2 infection in and 3% and 0.5%, respectively. With sodium chloride there was a significantly higher relative risk for positive cultures (1.7) and for the presence of Staphylococcus aureus (3.3). The chlorhexidine group required significantly fewer antibotics reported significantly less pain at weeks 6 and 10. CONCLUSIONS: Chlorhexidine solution (2 mg/ml) as cleansing agent in pin site care is preferable to sodium chloride in patients operated on by the hemicallotasis technique. PMID- 15338245 TI - Role of force--frequency relation during AV-block, sinus node block and beta adrenoceptor block in conscious animals. AB - OBJECTIVE: Myocardial contractility is regulated by adrenergic stimulation, the strength-length relationship and the force-frequency relationship or Bowditch effect. The latter mechanism was clearly demonstrated in muscle strips, in the isolated heart as well as in in-vivo experiments. The aim of this study was to further investigate the role of the force-frequency effect on the contractile response to exercise or isoproterenol infusion in conditions of restricted increases in heart rate i.e., AV-block, sinus node block and beta-adrenoceptor block. METHODS: Nineteen dogs were instrumented with a left ventricular miniature pressure gauge, catheters in the aorta, pulmonary artery and left atrium and pacing leads on the left atrium and left ventricle. In order to control the chronotropic response during sympathetic stimulation, permanent AV-block was induced in nine dogs, sinus node block using UL-FS 49 and beta-adrenoceptor block using propranolol was studied in ten dogs. RESULTS: Adrenergic stimulation (isoproterenol 0.4 micro g/kg or exercise) after total AV-block failed to increase LVdP/dt. However, increasing LV pacing rate (from 50 up to 200 bpm) prior to adrenergic stimulation elicited a significant increase in LVdP/dt (4762 +/- 166 mmHg/s vs. 6485 +/- 381 mmHg/s, p < 0.05). In dogs in sinus rhythm, heart rate and LVdP/dt response to isoproterenol and exercise following pre-treatment with UL-FS 49 is significantly reduced, with heart rate increasing from 103 +/- 7 up to 154 +/- 5 bpm and LV dP/dt(max) from 2925 +/- 171 mmHg/s to 6249 +/- 400 mmHg/s compared to the response in control conditions (HR 220 +/- 3 bpm and LV dP/dt(max) 7473 +/- 616 mmHg/s) (p < 0.05). When heart rate is matched using atrial pacing, the LVdP/dt(max) response reached comparable values as observed in control conditions (7310 +/- 550 mmHg/s). Similar responses were obtained during exercise. Beta-adrenoceptor blockade attenuates considerably the heart rate and LVdP/dt response to sympathetic stimulation. Adjusting heart rate with atrial pacing restores only partially LVdP/dt(max). CONCLUSION: During sympathetic stimulation, the chronotropic response plays a major role for the concomitant full expression of the inotropic response. In conditions where increases in heart rate are absent or severely restricted such as in permanent AV-block, sinus node block and beta-adrenoceptor block, the inotropic response will also be impaired. PMID- 15338246 TI - Regional dietary habits of French women born between 1925 and 1950. AB - BACKGROUND: Diseases distributions are not the same all over France. As diet is an important determinant of health it is essential to determine whether there was still a diversity in food habits across French regions. AIM OF THE STUDY: We examined regional differences in dietary habits and nutrient intakes among French women born between 1925 and 1950 participants in the "Etude Epidemiologique aupres des femmes de l'Education Nationale" (E3N) cohort. METHODS: Data were extracted from self-administered dietary history questionnaires completed by 73024 highly educated, middle-aged women between 1993 and 1995. Canonical and cluster analyses were used to identify contiguous areas of homogeneous dietary habits spanning two or more of the 20 French regions. Dietary clusters were described relatively to the entire cohort mean. RESULTS: Eight dietary clusters were identified. The following food items were overconsumed: cooked vegetables in the Mediterranean, fish in the West, fruit in the South-East, and potatoes in the North. The following food items were under-consumed: fish in the East, fruit in the North, and potatoes in the South-East and Mediterranean cluster. Consumption of soup and fruit increased with age, while consumption of pork, horse meat and coffee fell with age. Ethanol intake was highest in the North and lowest in the South-East; the types of alcoholic beverages consumed also varied across clusters. Total energy intake, nutrient intakes, and the contribution of carbohydrates, fat and protein to total energy intake were similar across clusters. Intake of cholesterol and polyunsaturated fatty acids varied across clusters. CONCLUSION: Dietary habits and alcohol consumption show marked regional differences in this population of middle-aged, highly educated French women. Changes in dietary behaviour with age involved few food items and were similar across clusters, suggesting that regional differences in food and beverage consumption persist. PMID- 15338247 TI - Weight of evidence needed to substantiate a health effect for probiotics and prebiotics: regulatory considerations in Canada, E.U., and U.S. AB - Successful and responsible introduction of probiotic and prebiotic products into the worldwide marketplace requires labelling for health benefits that meets consumer needs, adheres to regulatory standards and does not overextend scientific evidence. Regulations differ among countries, but underlying all is an emphasis on scientific credibility of any statements of health benefits. This paper considers the value of different types of evidence offered in substantiation of efficacy and reviews different regulatory approaches to labelling for health claims. Limitations of in vitro, animal and different types of human studies used for efficacy substantiation for probiotics and prebiotics are discussed. PMID- 15338248 TI - Low bone mineral density and bone mineral content are associated with low cobalamin status in adolescents. AB - BACKGROUND: Cobalamin deficiency is prevalent in vegetarians and has been associated with increased risk of osteoporosis. AIM OF THE STUDY: To examine the association between cobalamin status and bone mineral density in adolescents formerly fed a macrobiotic diet and in their counterparts. METHODS: In this cross sectional study bone mineral density (BMD) and bone mineral content (BMC) were determined by DEXA in 73 adolescents (9-15 y) who were fed a macrobiotic diet up to the age of 6 years followed by a lacto-(-ovo-) vegetarian or omnivorous diet. Data from 94 adolescents having consumed an omnivorous diet throughout their lives were used as controls. Serum concentrations of cobalamin, methylmalonic acid (MMA) and homocysteine were measured and calcium intake was assessed by questionnaire. Analysis of covariance (MANCOVA) was performed to calculate adjusted means for vitamin B12 and MMA for low and normal BMC and BMD groups. RESULTS: Serum cobalamin concentrations were significantly lower (geometric mean (GM) 246 pmol/L vs. 469 pmol/L) and MMA concentrations were significantly higher (GM 0.27 micromol/L vs. 0.16 micromol/L) in the formerly macrobiotic-fed adolescents compared to their counterparts. In the total study population, after adjusting for height, weight, bone area, percent lean body mass, age, puberty and calcium intake, serum MMA was significantly higher in subjects with a low BMD (p = 0.0003) than in subjects with a normal BMD. Vitamin B12 was significantly lower in the group with low BMD (p = 0.0035) or BMC (p = 0.0038) than in the group with normal BMD or BMC. When analyses were restricted to the group of formerly macrobiotic-fed adolescents, MMA concentration remained higher in the low BMD group compared to the normal BMD group. CONCLUSIONS: In adolescents, signs of an impaired cobalamin status, as judged by elevated concentrations of methylmalonic acid, were associated with low BMD. This was especially true in adolescents fed a macrobiotic diet during the first years of life, where cobalamin deficiency was more prominent. PMID- 15338249 TI - Lyophilized carrot ingestion lowers lipemia and beneficially affects cholesterol metabolism in cholesterol-fed C57BL/6J mice. AB - BACKGROUND: Several lines of evidence indicate that diet rich in fruit and vegetable can protect against cardiovascular diseases by acting on cholesterol metabolism and on oxidative stress. AIM OF THE STUDY: The aim of this study was to assess whether daily carrot consumption (provided as lyophilized powder) could differentially influence the consequences of cholesterol supplementation on lipid metabolism and oxidative stress in C57BL/6J mice. METHODS: Fourteen mice were randomized in four groups. Mice were fed either control diets (without or with 0.25% cholesterol added) or lyophilized carrot enriched diets (20% wt/wt without or with 0.25 % cholesterol added) for 4 weeks. Cholesterol and triglycerides in plasma and in liver were measured at the end of the experimental period. Fecal excretion of sterols was evaluated. Vitamin E and carotenoid concentrations were also determined. Several biomarkers relative to oxidative stress such as FRAP (Ferric Reducing Ability of Plasma) and isoprostanes were investigated. RESULTS: Feeding the carrot diet resulted in a decrease of cholesterol (-41%) and triglycerides (-49 %) in plasma and in the liver (-41% and -39%, respectively) in animals fed cholesterol-supplemented diets. Carrot diet induced an increase of total neutral sterols fecal excretion, which inhibits digestive cholesterol absorption. Carrot diet increased antioxidant status in cholesterol-fed mice as related by the 16% higher FRAP values. Although vitamin E was not affected by carrot diet, vitamin E/TG ratio was significantly higher in animals fed carrot diets. The carrot diet induced an increase of vitamin E in the heart in both cholesterol-free and cholesterol-supplemented mice suggesting a higher protection of this tissue. CONCLUSION: This study shows that carrot ingestion decreases lipemia and improves antioxidant status in mice. Such results suggest that carrot intake may exert a protective impact against CVD linked to atherosclerosis. It is likely that these effects could be due to the synergistic effect of fiber and associated antioxidants. PMID- 15338252 TI - [Radiological diagnostics of cervical rheumatoid arthritis]. AB - Rheumatoid arthritis leads to characteristic findings at the synovial joints, the intervertebral discs and the processes of the cervical spine. Isolated findings are not specific for rheumatoid arthritis. In fact, due to common underlying pathophysiologic changes they also develop in other inflammatory diseases affecting the cervical spine. Therefore, each radiological examination is to be understood and used as a piece in the diagnostic puzzle. Only in conjunction with clinical information does it add to a conclusive diagnosis. Nevertheless certain patterns of findings help in narrowing the list of differential diagnosis. Besides their role in initial diagnosis, radiological examinations are crucial tools in the peri- and post-operative work-up and in the detection of typical complications of rheumatoid arthritis with cervical manifestations, i. e. various instabilities and their consequences, as those have an impact on the therapeutic approach and prognosis. PMID- 15338253 TI - [Therapy of cervical rheumatoid arthritis]. AB - The rheumatoid involvement of the cervical spine can be divided into three phases. In the early stage of the disease there is an isolated atlantoaxial subluxation (AAS), followed by vertical instability and subaxial instability. If patients show clear symptoms of cervical myelopathy, which can occur during any stage of the disease, the progression cannot be stopped by conservative treatment, which is of great importance at the beginning of the cervical manifestation. Patient education, physiotherapy and immobilization with a stiff collar can significantly reduce pain. Early and effective DMARD therapy can have a positive effect on the natural history of the disease. In case of progressive instability, cervical myelopathy or severe pain operative treatment is indicated. If there is an isolated AAS, fusion can be restricted to the C1/C2 segment. The Magerl transarticular screw fixation is the preferred technique for stabilization. If there is evidence for vertical instability or severe destruction of the C0/C1 joints, occipital cervical fusion has to be performed. Durin the preoperative planning it is necessary to look for signs of subaxial instability. If this is the case, fusion should include the entire cervical spine. Transoral decompression may be necessary when there is persistent anterior compression of the myelon, typically seen in fixed AAS. Non-ambulatory myelopathic patients are more likely to develop severe surgical complications. Therefore, it is important to avoid the development of severe cervical instability by early surgical intervention. The right timing for surgery is still a matter of controversy. Future prospective randomized trials should address this topic to improve the treatment concept for the rheumatoid patient. PMID- 15338251 TI - [Cervical myelopathy as a complication of rheumatoid arthritis]. AB - Arthritis of the cervical spine with instability of the atlantodental joint is a typical and frequent complicaiton in rheumatoid arthritis. Subsequent cervical myelopathy is rare but usually a severe complication. There is no stringent correlation between arthritis of cervical spine an occurrence of cervical myelopathy. Cervical myelopathy is often difficult to discern from parallel multilating peripheral joint damage, which makes the diagnosis difficult. The decision between conservative and operative therapeutic intervention has to be based on subtle clinical, neurophysiological and radiological assessment and-in particular-requires exact analysis of the course of the underlying rheumatoid arthritis and myelopathy. In this diagnostic process close collaboration of rheumatologist, surgeon and neurologist is essential. PMID- 15338254 TI - [Columnotomy in ankylosing spondylitis]. AB - Depending on its severity the fixed flexion deformity of the spine in patients with ankylosing spondylitis leads to significant disturbance in the activity of daily living. The surgical correction of such a deformity requires a special knowledge in the context of spinal surgery. Meticulous surgical procedures were especially developed for this kind of spinal deformity. Considering the rate of complications it is necessary to select carefully the patients undergoing these procedures. The surgeon must be familiar in handling these patients and these special surgical procedures. PMID- 15338255 TI - [Scleroderma and Paul Klee: Metamorphosis of life and art?]. AB - Paul Klee was born in 1879 in Munchenbuchsee, not far from Bern, and died at the age of 60 in Switzerland due to severe scleroderma, which was diagnosed in 1936. From this time on, this connective tissue disease had followed him continuously and had strongly influenced his enjoyment of life, personal development and creative power in art work, before finally causing his death in 1940. Based on diaries, letters, personal remarks and notes, his case history was reconstructed from the first symptoms to the date of diagnosis up until his death and an analysis was made of the influence his illness had on his art work in aura, color, figure and productivity. PMID- 15338256 TI - [Adaptive patterns of the rheumatoid wrist after radiolunate arthrodesis]. AB - OBJECTIVE: The aim of this study was the analysis of long-term carpal changes after radiolunate arthrodesis. METHODS: Pre- and postoperative X-rays of 91 wrists in 78 patients with rheumatoid arthritis, who were treated for carpal instability with a radiolunate arthrodesis, were examined concerning the midcarpal joint and the Larsen grade. The mean follow-up was 60 months. RESULTS: After radiolunate arthrodesis the midcarpal joint space remained unchanged in 28%. In 35% secondary arthritis and in 37% further arthritic destruction occurred. The mean Larsen grade increased from 3.2 to 3.8. Six wrists needed complete arthrodesis 25 to 87 months after the primary procedure and one was treated by total wrist replacement. Adaptive changes of the carpus during progressive disease and in secondary arthritis were recognized. Three types of joint lines could be identified: in 35% of the wrists a "perilunar", in 22% a "radio-midcarpal" and in 3% a purely "midcarpal" joint line. In 40% no specific joint line could be identified. CONCLUSION: After radiolunate arthrodesis the carpus remains unchanged in the long run in nearly a third of cases. But even if secondary arthritis or further arthritic destruction occurs as in the remaining cases, the carpus shows an amazing capacity for adaptation. A new intracarpal joint line may develop or the midcarpal joint re-establishes itself. PMID- 15338257 TI - [Oxidatively modified lipoproteins and their antibodies in patients with antiphospholipid syndromeand systemic lupus erythematosus]. AB - The antiphospholipid syndrome (APS) with its typical clinical manifestations of recurrent thrombosis and fetal loss is biochemically defined by the presence of circulating antiphospholipid antibodies (aPL). The disease pattern has raised special interest as a possible link between autoimmunity and atherosclerosis. aPL, oxidized low density lipoproteins (oxLDL), and antibodies to oxLDL (Anti oxLDL) are suggested to play an important role in atherogenesis. In the present study we compared the serum levels of oxLDL and Anti-oxLDL in APS patients (20 subjects with primary APS; 14 subjects with secondary APS) and nonAPS subjects (24 phenotypically healthy controls samples and 12 patients with systemic lupus erythematosus [SLE]) and investigated associations of the above mentioned parameters with the intima-media thickness (IMT), a clinical surrogate parameter of atherosclerosis.SLE patients with and without APS showed significantly increased levels of Anti-oxLDL as compared to the controls group (p = 0.038 and p = 0.007, respectively). In contrast, oxLDL levels were not significantly different between the controls group and patients. The Anti-oxLDL levels correlated significantly with anticardiolipin (p = 0.002) and beta(2) glycoprotein I antibodies (p < 0.048), both from IgG isotype. Only SLE patients without APS revealed a significantly elevated production of reactive oxygen species indicating an increased proatherogenic oxidative stress in the circulation (p < 0.002). In the patient groups, the circulating levels of oxLDL and Anti-oxLDL showed no association with atherosclerosis as estimated by IMT. In conclusion, our experimental data do not support the concept of oxidative stress induced accelerated atherosclerosis in APS patients. PMID- 15338258 TI - [Acute auricular chondritis in relapsing polychondritis]. PMID- 15338259 TI - [Churg-Strauss syndrome: outcome and long-term follow-up of 32 patients]. PMID- 15338260 TI - [Hans Reiter and the oculo-urethro-synovial syndrome. Epilogue and legacy]. PMID- 15338263 TI - Native root xylem embolism and stomatal closure in stands of Douglas-fir and ponderosa pine: mitigation by hydraulic redistribution. AB - Hydraulic redistribution (HR), the passive movement of water via roots from moist to drier portions of the soil, occurs in many ecosystems, influencing both plant and ecosystem-water use. We examined the effects of HR on root hydraulic functioning during drought in young and old-growth Douglas-fir [ Pseudotsuga menziesii (Mirb.) Franco] and ponderosa pine ( Pinus ponderosa Dougl. Ex Laws) trees growing in four sites. During the 2002 growing season, in situ xylem embolism, water deficit and xylem vulnerability to embolism were measured on medium roots (2-4-mm diameter) collected at 20-30 cm depth. Soil water content and water potentials were monitored concurrently to determine the extent of HR. Additionally, the water potential and stomatal conductance ( g(s)) of upper canopy leaves were measured throughout the growing season. In the site with young Douglas-fir trees, root embolism increased from 20 to 55 percent loss of conductivity (PLC) as the dry season progressed. In young ponderosa pine, root embolism increased from 45 to 75 PLC. In contrast, roots of old-growth Douglas fir and ponderosa pine trees never experienced more than 30 and 40 PLC, respectively. HR kept soil water potential at 20-30 cm depth above -0.5 MPa in the old-growth Douglas-fir site and -1.8 MPa in the old-growth ponderosa pine site, which significantly reduced loss of shallow root function. In the young ponderosa pine stand, where little HR occurred, the water potential in the upper soil layers fell to about -2.8 MPa, which severely impaired root functioning and limited recovery when the fall rains returned. In both species, daily maximum g(s) decreased linearly with increasing root PLC, suggesting that root xylem embolism acted in concert with stomata to limit water loss, thereby maintaining minimum leaf water potential above critical values. HR appears to be an important mechanism for maintaining shallow root function during drought and preventing total stomatal closure. PMID- 15338264 TI - Reduction in size and fecundity of the autumnal moth, Epirrita autumnata, in the increase phase of a population cycle. AB - Increasing fecundity with increasing density has been observed for many cyclic herbivore populations, including some forest Lepidoptera. We monitored population density, body size and reproductive capacity of the cyclic lepidopteran, the autumnal moth (Epirrita autumnata, Geometridae), from the early increase phase to the devastating outbreak density in northernmost Norway. Larval density of the species increased exponentially from 1998 to 2002 and remained at the outbreak level also in 2003. Within the same period, the body size and fecundity of individuals reduced as analysed from several parallel datasets on larvae, pupae and adults. In another study area in northernmost Finland, the density increase of the autumnal moth was moderate only, and true outbreak density was not attained during the study. Despite that, a reduction was again detected in the size and fecundity of individuals. Possible factors responsible for the reduced size and fecundity of individuals in the Norwegian population were quantitative shortage of foliage, rapid and delayed inducible resistances of the host, mountain birch (Betula pubescens ssp. czerepanovii), as well as crowding-induced responses of larvae. These factors likely acted in concert, although non-delayed responses to the density were emphasized. Our findings did not support the hypotheses of climatic release, inducible susceptibility of the host tree and mast depression (i.e. lowered chemical defence of the host tree after its mast seeding) as promoters of the fecundity-based density increase of the autumnal moth, since the reduced fecundity in relation to increased density was strongly against the predictions of these hypotheses. Therefore, we suggest that the density increase of autumnal moth populations is promoted by high survival rather than exceptionally high fecundity. PMID- 15338265 TI - Resolving within- and between-population variation in feeding ecology with a biomechanical model. AB - Studies of phenotypic plasticity have emphasized the effect of the environment on the phenotype, but plasticity can also be used as a tool to study the functional significance of key traits. By inducing variation in phenotypes and testing quantitative models that predict performance based on biological mechanisms, we can develop functionally general models of performance. Pumpkinseed sunfish from lakes with high snail availability have large levator posterior muscles (which are used to crush snail shells), whereas fish from lakes with few snails have relatively small muscles. Here we: (1) quantify differences in the feeding ability of an ontogenetic series of pumpkinseed from two populations; and (2) evaluate whether a biomechanical model can resolve the observed ontogenetic and between-population variation in feeding ecology. Mass, but not length, of the levator posterior muscle in fish from Three Lakes (a lake rich in snails) was greater than for comparably sized fish from Wintergreen Lake (a lake with few snails). Handling times were shorter, crushing strengths were 71% greater, and foraging rate (snail tissue mass consumed per time) and the fraction of thick shelled snails in the diet were approximately 100% greater for fish from Three Lakes compared to comparably sized fish from Wintergreen. These between-lake differences were not significant after adjusting for variation in pharyngeal morphology, suggesting that the biomechanical model of snail crushing resolved observed ontogenetic and population-level variation in the feeding ecology of pumpkinseed. PMID- 15338266 TI - Links between the structure of an Antarctic shallow-water community and ice-scour frequency. AB - Ice is a major structuring force in marine and freshwater environments at high latitudes. Although recovery from scouring has been quantified in time, the frequency of scouring in the Antarctic has not. We placed grids of markers at 9 17 m depth at two sites, to study ice-scouring over 2 years at Adelaide island (Antarctic Peninsula). We quantified the time scale of scour frequencies, and linked this to community mortality, age and diversity. Markers were hit from zero to at least three times in 2 years. At the least disturbed site (South Cove) 24% of markers were destroyed per year, whereas in North Cove 60% of markers were destroyed. There were significant differences in scouring frequency between our two sites: a given area in North Cove was on average hit twice as often as one in South Cove. Compared with near shore environments elsewhere, faunas of both sites were characteristic of high disturbance regimes, exhibiting low percent cover, diversity, ages and a high proportion of pioneers. Aspects of the encrusting communities studied reflected the differences between site disturbance regimes. North Cove was scoured twice as often, and bryozoan communities there had half the number of species, two-thirds the space occupation and twice the mortality level of those in South Cove. Maximum age in North Cove bryozoans was also half that in South Cove. Although there are natural disturbance events that rival ice scouring in either frequency or catastrophic power at lower latitudes, none do both nor across such a wide depth range. We suggest that ice scour effects on polar benthos are even more significant than the same magnitude of disturbance at lower latitudes as recovery rates of high latitude communities are very slow. Climate warming seems likely to increase iceloading of near shore polar waters, so that some of the world's most intensely disturbed faunas may soon suffer even more disturbance. PMID- 15338267 TI - Induced defences in an endangered amphibian in response to an introduced snake predator. AB - Introduced species have contributed significantly to the extinction of endemic species on islands. They also create new selection pressures on their prey that may result in modified life history strategies. Introduced viperine snakes ( Natrix maura) have been implicated in the decline of the endemic midwife toad of Mallorca ( Alytes muletensis). A comparison of A. muletensis tadpoles in natural pools with and without snakes showed that those populations subject to snake predation possessed longer tails with narrower tail fins but deeper tail muscles. Field and laboratory experiments showed that these changes in tail morphology could be induced by chemical and tactile cues from snakes. Populations of tadpoles that were subject to snake predation also displayed clear bimodal size frequency distributions, with intermediate-sized tadpoles missing from the pools completely. Tadpoles in pools frequented by snakes developed faster in relation to their body size than those in pools without snakes. Variation in morphology between toad populations may therefore be caused by a combination of size selective predation and tadpole plasticity. The results of this study indicate that the introduction of alien species can result in selection for induced defences, which may facilitate coexistence between predator and prey under certain conditions. PMID- 15338268 TI - Characterisation of neurons expressing calbindin immunoreactivity in the ileum of the unweaned and mature sheep. AB - We have identified the enteric neuron types expressing immunoreactivity for the calcium-binding protein calbindin D28k (CALB) in cryostat sections and whole mount preparations of myenteric (MP) and submucosal (SMP) plexuses of sheep ileum. We wished to determine whether CALB-IR in the sheep enteric nervous system was expressed in Dogiel type II cells, as in guinea-pig and rat ileum, and could therefore be used as a marker for intrinsic primary afferent neurons. The neurochemical coding of CALB-containing myenteric and submucosal neurons in ileum of unweaned lamb and mature sheep and its co-localisation with various neural markers was studied immunohistochemically. An antiserum against neuronal nuclear protein (NeuN) failed to detect the entire neuronal population; it was expressed only in 48% of neuron-specific enolase (NSE)-immunoreactive (NSE-IR) neurons. Human neuronal protein appeared to occur in the large majority or all neurons. Almost all CALB-IR neurons were: (1) radially multidendritic; (2) eccentric multidendritic; (3) Dogiel type II. CALB-IR occurred in 20-25% of myenteric and 65-75% of submucosal neurons in lamb and mature sheep, with higher values in mature sheep. Nearly all CALB-IR neurons were common choline acetyltransferase (cChAT)-IR, whereas only about 20% of cChAT-IR somata were CALB-IR. In lamb and mature sheep, 90% of MP CALB-IR neurons were peripheral choline acetyltransferase (pChAT)-IR. In lamb SMP, 80+/-13% of CALB-IR cells were also pChAT-IR, whereas all those in mature SMP were pChAT-IR. Fewer myenteric CALB-IR neurons exhibited tachykinin (TK) in mature sheep (49%) than in lamb (88%). This was also the case for submucosal ganglia (mature sheep, 63%; lamb, 89%). In lamb MP, 77+/-7% of CALB-IR cells were NeuN-positive. In mature sheep, 73+/-10% of CALB-IR somata were NeuN-IR, but NeuN failed to stain SMP neurons. In the MP of suckling and mature sheep, Dogiel type II CALB-IR neurons were calcitonin gene-related peptide (CGRP)-IR. In the SMP at both stages, Dogiel type II CALB-IR somata (about 50% of CALB-IR neurons) were also CGRP-IR. Only small proportions of CALB-IR neurons showed immunoreactivity for calretinin or nitric oxide synthase (NOS), although large populations of CALB and NOS neurons occurred in the ganglia. Thus, CALB is a marker of most Dogiel type II neurons in the sheep but is not confined to Dogiel II neurons. CGRP is a more selective marker of Dogiel type II neurons, being only found in this neuron type. PMID- 15338269 TI - Innervation of the substantia nigra. AB - This review describes inputs to neurons in the substantia nigra and contrasts them with the action of agonists for the putative receptors through which they act. Special emphasis is placed on gamma-aminobutyric acid (GABA) afferents. Dopamine released from the somato-dendritic compartment of dopamine neurons and endocannabinoids released from dopamine and GABA neurons serve as retrograde signals to modulate GABA release. The release may be fostered by Ca(2+) release from intracellular Ca(2+) stores, which in turn may be influenced by the inputs. PMID- 15338270 TI - Aquaporin expression is downregulated in a murine model of colitis and in patients with ulcerative colitis, Crohn's disease and infectious colitis. AB - Colitis is associated with alterations in electrolyte and water transport. These changes give rise to some of the symptoms experienced by patients with colitis. Alterations in fluid flux may also contribute to increased susceptibility to mucosal injury. Recently, endogenous water channel proteins (aquaporins; AQPs), have been identified in colonic tissue. The expression of AQP4, AQP7 and AQP8 was examined, via reverse transcription/polymerase chain reaction, Western blotting and immunohistochemistry, in a murine model of colitis and in patients with inflammatory bowel disease or infectious colitis. Colitis was induced in C57BL/6 mice by the addition of 2.5% dextran sodium sulphate (DSS) to their drinking water. AQP expression in these mice was assessed following 12 h to 7 days of DSS exposure and during the recovery phase from 1 to 15 days following cessation of DSS exposure. Colonic water transport was measured after 1 and 3 days of DSS and following 7 days of recovery. The expression of AQP4 and AQP8 mRNA was significantly decreased after 12-24 h of DSS exposure and remained depressed throughout the treatment period. Expression of AQP7 was more variable. Protein expression followed a similar pattern to that observed for AQP mRNA. Significant alteration in colonic fluid secretion was correlated with reduced expression of AQP isoforms. Significantly, patients with active ulcerative colonic, Crohn's colitis or infectious colitis had similar dramatic reductions in AQP expression that appeared to be correlated with disease activity. Thus, colonic injury in both mouse and man is associated with a downregulation in AQP expression. PMID- 15338271 TI - Cellular pathology of Parkinson's disease: astrocytes, microglia and inflammation. AB - Parkinson's disease (PD) is a frequent neurological disorder of the basal ganglia, which is characterized by the progressive loss of dopaminergic neurons mainly in the substantia nigra pars compacta (SNpc). Inflammatory processes have been shown to be associated with the pathogenesis of PD. Activated microglia, as well as to a lesser extent reactive astrocytes, are found in the area associated with cell loss, possibly contributing to the inflammatory process by the release of pro-inflammatory prostaglandins or cytokines. Further deleterious factors released by activated microglia or astrocytes are reactive oxygen species. On the other hand, they may mediate neuroprotective properties by the release of trophic factors or the uptake of glutamate. In this review, we will discuss the different aspects of activated glial cells and potential mechanisms that mediate or protect against cell loss in PD. PMID- 15338272 TI - Stages in the development of Parkinson's disease-related pathology. AB - The synucleinopathy, idiopathic Parkinson's disease, is a multisystem disorder that involves only a few predisposed nerve cell types in specific regions of the human nervous system. The intracerebral formation of abnormal proteinaceous Lewy bodies and Lewy neurites begins at defined induction sites and advances in a topographically predictable sequence. As the disease progresses, components of the autonomic, limbic, and somatomotor systems become particularly badly damaged. During presymptomatic stages 1-2, inclusion body pathology is confined to the medulla oblongata/pontine tegmentum and olfactory bulb/anterior olfactory nucleus. In stages 3-4, the substantia nigra and other nuclear grays of the midbrain and forebrain become the focus of initially slight and, then, severe pathological changes. At this point, most individuals probably cross the threshold to the symptomatic phase of the illness. In the end-stages 5-6, the process enters the mature neocortex, and the disease manifests itself in all of its clinical dimensions. PMID- 15338273 TI - First case of aplastic anemia in a Japanese child with a homozygous missense mutation in the NBS1 gene (I171V) associated with genomic instability. AB - The NBS1 gene is strongly linked to several factors involved in genome integrity. Functional disruption of NBS1 could therefore induce genomic instability and carcinogenesis. Four children with acute lymphoblastic leukemia have been reported to be heterozygous for a germline and/or somatic missense mutation in NBS1, leading to the I171V substitution. We screened healthy controls and pediatric patients with hematological malignancies and aplastic anemia (AA) for the presence of I171V. Of the 62 patients, one individual with AA was confirmed to harbor a homozygous I171V mutation. Genetic analysis of NBS1 in this patient and her healthy parents indicated that she inherited the germline I171V mutation from her father and the wild-type allele from her mother, and that the second I171V hit occurred on the wild-type allele early in embryonic development. Furthermore, cytogenetic analysis of lymphoblastic cell lines from the patient indicated a remarkable increase in numerical and structural chromosomal aberrations in the absence of clastogens, suggesting that she potentially carried genomic instability. This is the first report of AA with a homozygous I171V mutation. We hypothesize that NBS1 may play an important role in the pathogenesis of AA. PMID- 15338274 TI - Analysis of genes implicated in iron regulation in individuals presenting with primary iron overload. AB - Extensive investigation into the molecular basis of iron overload disorders has provided new insights into the complexity of iron metabolism and related cellular pathways. The possible involvement of genes affecting iron homeostasis, including HFE, SLC40A1, HAMP and CYBRD1, was investigated in individuals who were referred for confirmation or exclusion of a diagnosis of haemochromatosis, but who tested negative or were heterozygous for the causative HFE mutation, C282Y. Denaturing high performance liquid chromatography analysis of these genes revealed a unique spectrum of mutations in the South African study population, including 67 unrelated patients and 70 population-matched controls. Two novel CYBRD1 gene mutations, R226H and IVS1-4C-->G, were identified in 11% of South African Caucasian patient referrals. We identified a novel D270V mutation in the SLC40A1 gene in a Black South African female with iron overload. These mutations were absent in the control population. In Africans with iron overload not related to the HFE gene, the possible involvement of the SLC40A1 and CYBRD1 genes was demonstrated for the first time. This study confirms the genetic heterogeneity of haemochromatosis and highlights the significance of CYBRD1 mutations in relation to iron overload. PMID- 15338275 TI - Low prevalence of MYOC mutations in UK primary open-angle glaucoma patients limits the utility of genetic testing. AB - Primary open angle glaucoma (POAG) affects 1% of people over age 40. Early detection and treatment can prevent blindness, but the disease is often asymptomatic until a late stage. Positive family history is an important risk factor and previous studies indicate that approximately 5% of POAG results from mutations in the myocilin ( MYOC) gene, raising the possibility of identifying individuals genetically predisposed to glaucoma. We collected DNA samples from 426 unselected UK POAG patients and analyzed them for MYOC mutations. The Q368X mutation was found in six patients (1.4%). No other mutations were identified, suggesting that amongst patients unselected for family history, the prevalence of MYOC mutations in the UK is lower than in other populations. Genetic and glaucoma screening was offered to first-degree relatives of these six probands (group 1) and of age/sex-matched mutation-negative controls (group 2). Of 11 group-1 relatives, three carried Q368X, one of whom already had glaucoma. Notably, of the 13 relatives in both groups who were mutation negative, one was already being treated for ocular hypertension. We therefore caution against changing glaucoma surveillance regimens in such individuals and suggest that routine untargeted genetic testing for MYOC mutations in patients with POAG would be of limited value until additional significant genetic risk factors are identified. PMID- 15338276 TI - Functional analysis of two-amino acid substitutions in gp91 phox in a patient with X-linked flavocytochrome b558-positive chronic granulomatous disease by means of transgenic PLB-985 cells. AB - Chronic granulomatous disease (CGD) is a rare inherited disorder in which phagocytes lack NADPH oxidase activity. The most common form is caused by mutations in the CYBB gene encoding gp91 phox protein, the heavy chain of cytochrome b(558), which is the redox element of NADPH oxidase. In some rare cases, the mutated gp91 phox is normally expressed but no NADPH oxidase can be detected. This type of CGD is called X91(+) CGD. We have previously reported an X(+) CGD case with a double-missense mutation in gp91 phox. Transgenic PLB-985 cells have now been made to study the impact of each single mutation on oxidase activity and assembly to rule out a possible new polymorphism in the CYBB gene. The His303Asn/Pro304Arg gp91 phox transgenic PLB-985 cells exactly mimic the phenotype of the neutrophils of the X(+) CGD patient. The His303Asn mutation is sufficient to inhibit oxidase activity in intact cells and in a broken cell system, whereas in the Pro304Arg mutant, residual activity suggests that the Pro304Arg substitution is less devastating to oxidase activity than the His303Asn mutation. The study of NADPH oxidase assembly following the in vitro and in vivo translocation of cytosolic factors p47 phox and p67 phox has demonstrated that, in the double mutant and in the His303Asn mutant, NADPH oxidase assembly is abolished, although the translocation is only attenuated in Pro304Arg mutant cells. Thus, even though the His303Asn mutation has a more severe inhibitory effect on NADPH oxidase activity and assembly than the Pro304Arg mutation, neither mutation can be considered as a polymorphism. PMID- 15338277 TI - Functional disomy resulting from duplications of distal Xq in four unrelated patients. AB - Duplications involving the X chromosome, in which the duplicated region is not subject to inactivation, are rare. We describe four distal Xq duplications, in three males and one female, in which the duplicated X chromosomal material is active in all cells. The infantile phenotype bears some resemblance to that of the Prader-Willi syndrome, presenting with initial feeding difficulties, hypotonia and, sometimes, with cryptorchidism. However, the severity of the phenotype is not simply related to the size of the duplication and so variations in gene expression, gene disruption or position effects from breakpoints should be considered as explanations. We have compared the clinical, cytogenetic and molecular findings of our patients with those previously reported. This has enabled us to question the suggestion that duplication of the gene SOX3 is the cause of hypopituitarism and that duplication of Filamin A is the cause of bilateral periventricular nodular heterotopia/mental retardation syndrome (BPNH/MR). We have also narrowed the putative critical interval for X-linked spina bifida. PMID- 15338278 TI - A longitudinal study of X-inactivation ratio in human females. AB - We investigated the effect of aging on X chromosome inactivation by performing a longitudinal study in a population of 178 normal females. We examined X inactivation ratios (fraction of cells with the same X chromosome active) in two sets of peripheral blood DNA samples collected about two decades apart. We observed a strong correlation between the ratios of individual females at the two time points and found no significant difference between the two sets of measurements. These observations indicate that aging, per se (as opposed to being "aged"), has little effect on X-inactivation. However, we also found that several females who were older than 60 years of age at the time of the first measurement acquired significant changes in the X-inactivation ratio. We speculate that, if X inactivation skewing is a frequently acquired trait in older females, it is acquired as the result of a discontinuous or catastrophic process and is not the result of constant selection for or against hematopoietic stem cells with a particular X chromosome active. PMID- 15338279 TI - Identification of a 98-kb DNA segment containing the rice Eui gene controlling uppermost internode elongation, and construction of a TAC transgene sublibrary. AB - The recessive 'tall rice' phenotype associated with the mutation eui (elongated upper-most internode) is an important agronomic trait that has been introduced into hybrid rice to eliminate panicle enclosure in all types of male-sterile lines and produce good-quality seeds in high yield and at low cost. Based on our previous Eui mapping data, we conducted fine-structure mapping and positional cloning of the gene using an F2 population comprising more than 5000 individuals derived from a cross of the near-isogenic lines 307T (eui/eui) with the recurrent parent Zhenshan 97 (Eui/Eui). In total 45 CAPS (cleaved amplified polymorphic sequences) markers located within an interval of 14.5 cM were analyzed in the subpopulation of 1298 homozygous recessive plants. The resulting high-resolution map defined a 98-kb interval containing the Eui locus flanked by the markers M0387 and M01, and three markers were found to co-segregate with Eui. In order to facilitate the identification of the Eui gene, we used a transformation-competent artificial chromosome (TAC) vector to construct a set of contiguous TAC clones from the Nipponbare BACs (obtained from the Clemson University Genome Institute; CUGI) spanning this region. These clones can be used to streamline complementation testing. The markers tightly linked to the Eui locus can also be used in breeding male-sterile lines with the elongated uppermost internode. PMID- 15338281 TI - A survey of SL1-spliced transcripts from the root-lesion nematode Pratylenchus penetrans. AB - Plant-parasitic nematodes are important and cosmopolitan pathogens of crops. Here, we describe the generation and analysis of 1928 expressed sequence tags (ESTs) of a splice-leader 1 (SL1) library from mixed life stages of the root lesion nematode Pratylenchus penetrans. The ESTs were grouped into 420 clusters and classified by function using the Gene Ontology (GO) hierarchy and the Kyoto KEGG database. Approximately 80% of all translated clusters show homology to Caenorhabditis elegans proteins, and 37% of the C. elegans gene homologs had confirmed phenotypes as assessed by RNA interference tests. Use of an SL1-PCR approach, while ensuring the cloning of the 5' ends of mRNAs, has demonstrated bias toward short transcripts. Putative nematode-specific and Pratylenchus specific genes were identified, and their implications for nematode control strategies are discussed. PMID- 15338282 TI - An improved method for detection of Leishmania amastigotes by an antibody probe against the small subunit of leishmanial ribonucleotide reductase. AB - By taking advantage of an antibody raised against the small M2 subunit of ribonucleotide reductase of Leishmania that reacts with the enzyme in the nucleus of the parasite but does not cross-react with the same enzyme of the host macrophage, an improved fluorescence-staining method is developed for enumeration of leishmanial amastigotes inside the macrophage. The method offers an accurate and easy way of counting, compared with Giemsa staining. PMID- 15338283 TI - Cytokine and antibody production during the course of resolution in Plasmodium yoelii 17XL-infected BALB/c mice treated with febrifugine and isofebrifugine mixture from leaves of Hydrangea macrophylla var. Otaksa. AB - Cytokine and antibody production was investigated during the course of resolution of primary infection in Plasmodium yoelii 17XL-infected BALB/c mice treated with a mixture of febrifugine and isofebrifugine. The infected mice in an untreated control group showed a progressively increasing parasitemia, leading to mouse death. In contrast, infected mice given the mixture orally showed low parasitemia levels during administration. Following a transient increase in parasitemia in the bloodstream of the treated mice, no parasites could be detected by microscopic examination. Analysis of cytokines in plasma showed that the plasma IFN-gamma levels elevated significantly within the first week of infection in both groups. Furthermore, on day 20 the plasma IFN-gamma and IL-4 levels elevated significantly in the treated mice and the production of both cytokines was sustained until at least day 40. The production of both cytokines in the treated mice was coincident with a decrease in parasitemia. The production of parasite specific antibodies in the course of P. yoelii 17XL infection was also monitored. In the drug-treated mice, the titers of parasite-specific IgG1, IgG2a, IgG2b and IgG3 elevated significantly from day 20; and the production of parasite-specific antibodies was coincident with a decrease in parasite numbers in the bloodstream. PMID- 15338280 TI - Comparative analysis of Mutator -like transposases in sugarcane. AB - The maize Mutator ( Mu) system has been described as the most active and mutagenic plant transposon so far discovered. Mu -like elements (MULEs) are widespread among plants, and many and diverse variants can coexist in a particular genome. The autonomous regulatory element MuDR contains two genes: mudrA encodes the transposase, while the function of the mudrB gene product remains unknown. Although mudrA -like sequences are ubiquitous in plants, mudrB seems to be restricted to the genus Zea. In the SUCEST (the Brazilian Sugarcane EST Sequencing Project) database, several mudrA -like cDNAs have been identified, suggesting the presence of a transcriptionally active Mu system in sugarcane. Phylogenetic studies have revealed the presence in plants of four classes of mudrA -like sequences, which arose prior to the monocot/eudicot split. At least three of the four classes are also found in the progenitors of the sugarcane hybrid (Saccharum spp.), Saccharum officinarum and S. spontaneum. The frequency of putatively functional transposase ORFs varies among the classes, as revealed at both cDNA and genomic levels. The predicted products of some sugarcane mudrA like transcripts contain both a DNA-binding domain and a transposase catalytic site motif, supporting the idea that an active Mu system exists in this hybrid genome. PMID- 15338284 TI - Tissue stages of Hepatozoon canis in naturally infected dogs from Sao Paulo State, Brazil. AB - A total of 222 dogs were examined by blood smear examination and Hepatozoon canis infection was detected in 13 dogs (5.9%). Five H. canis-infected dogs were necropsied to observe tissue stages in the organs. Fragments of spleen, liver, lungs, heart, kidneys, lymph nodes, bone marrow and skeletal muscles were used to made touch-impression smears. No macroscopic lesions were found in the organs. Two dogs had gamonts within polymorphonuclear cells and schizonts in various stages of development within the spleen and the bone marrow. Nevertheless, no mature meronts were found. PMID- 15338285 TI - Fasciola hepatica: the developmental patterns of redial generations in naturally infected Galba truncatula. AB - A retrospective study on 1,211 snails naturally infected with Fasciola hepatica in central France was carried out to determine the numbers of full-grown sporocysts from which infections derived, to specify the developmental patterns of redial generations (normal or abnormal) and to count live and free rediae. In the department of Haute Vienne (siliceous subsoils), most snails showed single sporocyst infections, with normal (46.3%) or abnormal (33.9%) development of redial generations. Two-sporocyst infections were scarcer (a total of 10.6%), while snail co-infections with F. hepatica and Paramphistomum daubneyi (8.0%) were found since 1996. In the department of Indre (calcareous subsoils), single sporocyst infections showing normal development of rediae were the most numerous (58.0%), while the frequency of single-sporocyst infections with abnormal development was strongly decreased (4.3%). Two-sporocyst infections (with normal development of generations) and co-infections with F. hepatica and P. daubneyi slightly increased in frequency. The redial burdens found in snails collected from the department of Indre were significantly higher than those noted in snails originating from the department of Haute Vienne, whatever the type of snail infection; and these increases in numbers especially concerned the rediae of the second and subsequent generations. The results might be explained by the lower calcium ion content (7-23 mg/l) present in waters from the department of Haute Vienne which would induce a slower growth of infected snails (the shell height of adults scarcely reached 8 mm) and, consequently, would create barely favourable conditions for the development of the first rediae of the first generation within snails. However, the quality of the diet provided as food for snails and its influence on the development of redial generations cannot be excluded. PMID- 15338286 TI - Enhancement of oral moxidectin bioavailability in rabbits by lipid co administration. AB - Moxidectin is a member of the macrocyclic lactone family of drugs widely used for the control of internal and external parasites. Because moxidectin is highly lipophilic, we suspect that lymphatic transport influences the intestinal absorption of oral formulations of the drug. We studied the influence of lipid co administration on the pharmacokinetics of an oral formulation of moxidectin in rabbits. Ten rabbits were orally administered 0.3 mg kg(-1) moxidectin with or without sunflower oil. Moxidectin and triglyceride were analyzed in plasma over 23 days. Sunflower oil co-administration significantly increased the area under the plasma concentration-time curve of moxidectin (98%, P<0.05) and prolonged its mean residence time from 1.52 days to 2.12 days (P<0.04). Simultaneously, an increase in plasma triglyceride was observed in response to oil administration. It is suggested that lipid administration increases the systemic availability of oral moxidectin by enhancing the extent of intestinal lymphatic transport of the drug. Lipid-based formulations should improve the bioavailability of moxidectin in rabbits. PMID- 15338287 TI - Trypanosoma evansi in inbred and Swiss-Webster mice: distinct aspects of pathogenesis. AB - Trypanosoma evansi (Trypanosomatidae, Kinetoplastida) is a salivarian trypanosomatid that infects eight mammal orders spread over America, Europe and Asia. In Brazil, T. evansi is the etiological agent of "Mal de Cadeiras", a horse disease very often described in the region known as Pantanal do Mato Grosso. Few data concerning the genetic diversity and biology of subpopulations of T. evansi that circulate in Brazil are available. The factors that modulate the interaction of this parasite with its hosts also remain to be elucidated. Here we evaluated the course of experimental infection of six T. evansi isolates derived from domestic and wild animals in Swiss-Webster mice and three Mus musculus lineages. The follow-up included biological, immunological as well as biochemical and hematological parameters. The same isolates as well as three others were characterized by pulsed-field electrophoresis. Our results showed that T. evansi isolates displayed significant differences regarding behavior and morbidity patterns in the distinct mouse lineages. Nevertheless, these differences could not be correlated with pulsed-field electrophoresis profiles. Indeed, concerning this molecular marker, only microheterogeneity was observed. Moreover, we observed that the outcome of the infection is defined by both host genetic background and peculiarities (virulence factors) of the distinct T. evansi isolates. Anemia and hypoglycemia were the only features that could be observed in all mouse lineages, independently of the inoculated T. evansi subpopulation. In addition, our data also show that Mus musculus is a suitable model host for the study of the different pathogenetic features of T. evansi infection. PMID- 15338288 TI - On the substantial variation in serological responses in pigs to Sarcoptes scabiei var. suis using different commercially available indirect enzyme-linked immunosorbent assays. AB - Two trials were carried out to compare the use of different enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against Sarcoptes scabiei var. suis. In the first trial, we employed four tests and 70 selected sera from a closed pig farm with sarcoptic mange. The sera were taken from the breeding area as well as from the finishing unit and compared with skin scrapings. The SARCOPTES-ELISA 2001 yielded the most positive results (88.58%), followed by the ELISA of the National Veterinary Institute, Uppsala (70%), the Acar-Test P-ELISA (52.86%), skin scrapings (48.57%), and the CHEKIT Sarcoptest (30%). In the second trial, eight litters from infected sows were examined from weeks 1-12 of life using the CHEKIT Sarcoptest and the SARCOPTES-ELISA 2001. The presence of maternal antibodies was highest on day 7 in both ELISAs and could be detected until weeks 5-9 of life. Antibodies increased as a result of an active immune response between days 56 and 63 in the SARCOPTES-ELISA 2001 and between days 70 and 77 of life in the CHEKIT Sarcoptest. Significant differences between the first and second litters were observed. PMID- 15338289 TI - Immunohistochemical characterization of the initial stages of Naegleria fowleri meningoencephalitis in mice. AB - The initial stages of Naegleria fowleri meningoencephalitis in mice were immunohistochemically characterized following the first 8 h post-intranasal inoculation. The events found after 8 h were: (1) amebas in contact with the mucous layer of the olfactory epithelium, (2) numerous parasites eliminated by extensive shedding of the mucous layer, and (3) many organisms reaching the nasal epithelium. In contrast to other works, we observed that after 24 h, amebas invaded the epithelium, without evidence of the disruption of the nasal mucosa. In addition some trophozoites invading through the respiratory epithelium were observed, suggesting an additional invasion route. The inflammatory response detected was scarce until 30 h post-inoculation. After 96 h, the inflammatory response was severe in the olfactory bulb and brain, and the tissue damage great. Consequently, an inflammatory reaction may enhance tissue damage but apparently does not destroy amebas which seem to proliferate in the olfactory bulb. PMID- 15338290 TI - Balamuthia mandrillaris, an opportunistic agent of granulomatous amebic encephalitis, infects the brain via the olfactory nerve pathway. AB - Balamuthia mandrillaris is a free-living ameba and an opportunistic agent of lethal granulomatous amebic encephalitis (GAE) in humans and other mammals. Its supposed routes of infection have been largely assumed from what is known about Acanthamoeba spp. and Naegleria fowleri, other free-living amebae and opportunistic encephalitis agents. However, formal proof for any migratory pathway, from GAE patients or from animal models, has been lacking. Here, immunodeficient mice were infected with B. mandrillaris amebae by intranasal instillation, the most likely natural portal of entry. By means of classical and immunohistology, the amebae are shown to adhere to the nasal epithelium, progress along the olfactory nerves, traverse the cribriform plate of the ethmoid bone, and finally infect the brain. A similar invasion pathway has been described for N. fowleri. The data suggest that the olfactory nerve pathway is a likely route for natural infection of the brain by B. mandrillaris amebae. PMID- 15338291 TI - The involvement of an integrin-like protein and protein kinase C in amoebic adhesion to fibronectin and amoebic cytotoxicity. AB - Adherence of a pathogen to the host cell is one of the critical steps in microbial infections. Naegleria fowleri, a causative agent of primary amoebic meningoencephalitis in humans, is expected to interact with extracellular components of the host, such as fibronectin, in a receptor-mediated mode. In this study, we investigated the interaction between N. fowleri and fibronectin to understand its cytopathology. In binding assays using immobilized fibronectin, the number of amoebae bound to fibronectin was increased compared to the controls, and was dependent on the amount of coated fibronectin present. A fibronectin binding protein of 60 kDa was found in extracts of N. fowleri. Western blot and immunolocalization assays using integrin alpha(5)/FnR antibodies showed that a 60 kDa protein reacted with the antibodies in extracts of N. fowleri, which was localized on the surface of N. fowleri. Preincubation of N. fowleri with the integrin antibodies significantly inhibited amoebic binding to fibronectin and cytotoxicity to the CHO cells. Additionally, protein kinase C activity was detected in the extract of N. fowleri. When N. fowleri was pretreated with protein kinase C activator or inhibitor, the abilities of amoebic adhesion to fibronectin and cytotoxicity to the host cells were markedly affected compared to untreated amoebae. These results suggest that an amoebic integrin like receptor and protein kinase C play important roles in amoebic cellular processes in response to fibronectin. PMID- 15338292 TI - Human and animal fascioliasis in Mazandaran province, northern Iran. AB - The fascioliasis situation in humans and livestock of Iranian Mazandaran is analysed for the first time. Coprological studies showed 7.3% and 25.4% global prevalences in sheep and cattle, respectively. Studies in slaughterhouses indicate that sheep and cattle may be the main reservoir species, buffaloes may play local roles in the transmission, and goats and horses probably only participate sporadically. Morphometric studies by computer image analysis showed that forms intermediate between Fasciola hepatica and F. gigantica appear in addition to both species. A total of 107 infected humans were diagnosed during the 1999-2002 period, which suggests that fascioliasis may be widespread. The absence of differences in human fascioliasis among gender and age groups differs from other human endemic areas. Both human and animal infections show marked differences between western and eastern Mazandaran. Traditions in herbal condiments for human consumption, methods of animal husbandry and annual rainfall may explain the higher prevalences in western Mazandaran. PMID- 15338293 TI - Fasciola hepatica and Paramphistomum daubneyi: vertical distribution of metacercariae on plants under natural conditions. AB - Four experiments on the metacercariae of Fasciola hepatica and Paramphistomum daubneyi were carried out under natural conditions in order to study their vertical location on submerged plants and to determine whether simultaneous cercarial shedding of both digenea causes changes in the distributions of the metacercariae. These experiments were performed in experimental boxes, each containing six tufts of rushes. Most metacercariae (73.0%) of F. hepatica were found along the walls of boxes, while 81.5% of P. daubneyi metacercariae were found on rush stems. In the case of snails infected by either of the two digenea, 80.1% of F. hepatica cercariae encysted on submerged parts of rushes and of the box walls near the water surface (to a depth of 1 cm), whereas 73.0% of P. daubneyi metacercariae were found in the lower sections (from -4 to -7 cm). If snails dually infected with P. daubneyi and F. hepatica were used, the vertical distributions of the metacercariae were significantly different from those found for snails infected by either of the two digenea. If snails having 42-day old infections with F. hepatica and other snails with 70-day old infections with P. daubneyi were simultaneously introduced into the boxes, the frequency of F. hepatica cysts was significantly lower in the section located under the water surface (29% only), while the frequencies of metacercariae in the lower sections (from -1 to -5 cm) were increased. Some significant changes were also observed for the metacercariae of P. daubneyi. The disturbance noted in the vertical distributions of F. hepatica and P. daubneyi metacercariae suggest that the encystment of F. hepatica cercariae can be disturbed by the simultaneous encystment of P. daubneyi cercariae, or conversely. PMID- 15338294 TI - The influence of zinc in mice on infection with Angiostrongylus cantonensis. AB - BALB/cByJ mice were divided into zinc-deficient (ZD), high zinc-supplemented (ZH), adequately zinc-replete (ZA) and normal control groups by daily dietary control. The body weight in ZD and ZH mice became significantly less than that of normal control mice from 4 weeks (P < 0.002) until 7 weeks (P < 0.0001) after consecutive zinc control. The kinetics of change in body weight of ZD and ZH mice after infection with Angiostrongylus cantonensis were similar to that of uninfected groups mentioned above. The worms recovered at days 7, 14 and 21 after infection from ZD and ZH mice were significantly more in number than that in control mice (P < 0.01). However, the worms from day 21 after infection were shorter in size than those in control mice (P < 0.01). The production of interleukin (IL)-5 was significantly depressed in cultured spleen cells from uninfected and infected ZD and ZH mice, compared with that from respective control group (P < or = 0.02). Furthermore, significantly lower eosinophil counts were observed in the cerebral spinal fluid(CSF) of ZD and ZH mice 3 weeks after infection than in the CSF of control mice (P < 0.001). The levels of immunoglobulins IgG, IgM, IgA and IgE in the supernatant of cultivated spleen cells and serum from ZD and ZH mice were all lower than those of control mice. Thus, the level of zinc correlated with the defence against infection by A. cantonensis, due to the influence of zinc on the growth of mice and their production of IL-5, eosinophils and immunoglobulins. PMID- 15338296 TI - Thrombocytopenia associated with second-line carboplatin-based chemotherapy for ovarian, fallopian tube, and primary peritoneal cavity cancers. AB - PURPOSE: There are limited data available in the oncology literature regarding the risk of thrombocytopenia associated with carboplatin-based second-line treatment of ovarian cancer, outside the setting of patients participating in clinical trials. METHODS: To examine this clinically-relevant issue, we conducted a retrospective review of the medical records of women with ovarian cancer treated in the Gynecologic Cancer Program of the Cleveland Clinic from 1994 through November 2003, who received >2 second-line carboplatin-based regimens. RESULTS: A total of 176 second-line carboplatin-based programs were delivered to the 152 patients (median age 61 years; range 39-87 years) identified through this review. A total of ten (7%) patients experienced >grade 2 thrombocytopenia, with only four (3%) patients developing platelet count nadirs <50.0 x 10(9)/l. No patient required discontinuation of platinum-based therapy due to the development of thrombocytopenia. CONCLUSION: Second-line carboplatin-based chemotherapy of ovarian cancer is associated with a low incidence of serious thrombocytopenia, if a strategy of selecting modest initial dose levels, and instituting rapid reductions in dose with the development of significant bone marrow suppression, is employed. PMID- 15338297 TI - Further delineation of El-Shanti syndrome. PMID- 15338298 TI - Early onset Huntington disease: a neuronal degeneration syndrome. AB - Huntington disease (HD) is an autosomal dominant, lethal neurodegenerative disorder of the central nervous system, caused by an uncontrolled expansion of a CAG dynamic mutation in the coding region of the IT15gene. Although a majority of patients have a midlife onset of the disease, in a small number of patients the disease manifests before 20 years of age. In adults, HD is mainly characterised by involuntary movements, personality changes and dementia. By contrast, in children a dominant picture of bradykinesia, rigidity, dystonia and epileptic seizures is noticed. The earlier onset is often associated with a paternal transmission of the disease allele to the offspring. We report here a rather unusual infantile onset of the disease in a little girl who presented with a history of seizures and psychomotor regression starting at the age of 3 years. A progressive cortical-subcortical atrophy, progressive cerebellar atrophy and lesions in the basal ganglia were found on MRI. An important expansion, of 214 triplet numbers, of the CAG repeat size associated with HD, was observed. CONCLUSION: Juvenile Huntingdon disease should be considered in children suffering from a progressive neurodegenerative disease. PMID- 15338299 TI - Increased early local immune responses and altered worm development in high-dose infections of mice susceptible to the filaria Litomosoides sigmodontis. AB - The relationship between the number of larvae inoculated and filarial infection outcome is an important fundamental and epidemiological issue. Our study was carried out with BALB/c mice infected with the filaria Litomosoides sigmodontis. For the first time, an immunological analysis of infection with various doses was studied in parallel with parasitological data. Mice were inoculated with 200, 60 or 25 infective larvae (third stage larvae, L3), and monitored over 80 days. At 60 h post-inoculation the immune response was stronger in the 200 L3 group than the 25 L3 group. Cells from lymph nodes draining the site of inoculation proliferated intensely and produced large amounts of IL-5 and IL-4. In the pleural cavity, leukocyte populations accumulated earlier and in larger quantities. IgG1, IL-4 and IL-10 serum concentrations were transiently higher. During the first 10 days the worm recovery rates were identical in all groups, but decreased thereafter in the 200 L3 group. In this group, the development of the worms was altered, with reduced lengths, diminished intra-uterine production of microfilariae and abnormalities of male copulatory organs. Whereas mice inoculated with 25 L3 became microfilaraemic, only one third reached patency in the 200 L3 group. However, detrimental effects of high numbers of worms are not seen in studies using different inoculation protocols. This suggests that the very early events determine subsequent immune response and infection outcome rather than competitive interactions between the worms. PMID- 15338300 TI - Premature regression of the leg apical ectodermal ridge in the Japanese chick wingless mutant. AB - The autosomal dominant Japanese wingless mutant has varying degrees of wing and leg truncations. The wing defects range from complete loss to negligible defects, whereas leg abnormalities are usually restricted to loss of the phalanges. Further analyses of the mutant focusing on the leg, which has been relatively uncharacterized, were performed. The expression pattern of Fgf8, a marker gene for the apical ectodermal ridge (AER) that controls outgrowth of the limbs, revealed premature regression at stage 28. Electron microscopy study showed abnormalities in the basement membrane all through the AER in the same stage. In the mutant, cell death was observed in the mesenchyme underlying AER between stages 31 and 32, although in the wild-type leg, AER regression and cell death occurred almost simultaneously at stages 33-34. To know if the cell death and cessation of the outgrowth are common mechanisms of wild-type and the mutant, we removed the AER in wild-type embryos at stage 28 and followed the fate of the limb. This also resulted in premature cell death 48 h after AER removal (equivalent to stage 32) and limb truncations similar to those observed in mutant limbs. To confirm whether either AER or underlying mesenchyme is responsible for the truncation, transplantation of the AER between the wild-type and the mutant was performed. This revealed that AER is the defective tissue in this mutant. PMID- 15338301 TI - Pelvic development in the rabbit embryo: implications in the organogenesis of bladder exstrophy. AB - Current theses of the development of bladder exstrophy and its variants rely on defective evolution of the urinary tract and cloacal membrane. They do not account satisfactorily for the clinical features reported in children affected by exstrophy, especially the pelvic bone anomaly. We herein describe the normal development of the pelvic ring in the rabbit embryo and its chronological relationship with the lower urinary tract organogenesis. Our results suggest that these events are intricated and allow us to propose a novel mechanism to explain exstrophies. PMID- 15338302 TI - Intrinsic cartilage-forming potential of dermomyotomal cells requires ectodermal signals for the development of the scapula blade. AB - The avian scapula has a dual origin. The cranial part derives from the somatopleure of the forelimb field, while the caudal part, the scapula blade, originates from the dermomyotomes of the cervicothoracic transition zone. Thus, these dermomyotomes have, in addition to the well-known myogenic, angiogenic, and dermogenic potential, the ability to form cartilage. The scapula blade is therefore a derivative of dermomyotomal chondrogenesis. Although the mechanisms that direct the sclerotomal chondrogenesis are beginning to be understood, little is known about dermomyotomal chondrogenesis. Here, we address the mechanisms that control dermomyotomal cells to become chondrocytes. After heterotopic transplantation of dorsal epithelial somite halves from the scapula-forming level to the cervical level, the grafted tissue retains the capability to form cartilage, indicating that the dermomyotomal chondrogenic potential must be specified during anterior-to-posterior regionalization of the paraxial mesoderm. Furthermore, we show that signals from the ectoderm are required, allowing dermomyotome cells to express markers associated with the chondrogenic lineage. PMID- 15338303 TI - Formation and differentiation of the avian dermomyotome. AB - During somite maturation, the ventral half of the epithelial somite disintegrates into the mesenchymal sclerotome, whereas the dorsal half forms a transitory epithelial sheet, the dermomyotome, lying in between the sclerotome and the surface ectoderm. The dermomyotome is the source of most of the mesodermal tissues in the body, giving rise to cell types as different as muscle, connective tissue, endothelium, and cartilage. Thus, the dermomyotome is the most important turntable of mesodermal cell fate choice in the vertebrate embryo. Here, we discuss the current knowledge on the formation of the dermomyotome and the mechanisms leading to the development of the various dermomyotomal derivatives, with special emphasis on the development of musculature and dermis. PMID- 15338304 TI - Erdheim-Chester disease of the breast associated with Langerhans-cell histiocytosis of the hard palate. AB - We report a patient with Langerhans-cell histiocytosis (LCH) localized to the hard palate that was later proven to be associated with Erdheim-Chester disease (ECD), involving the right breast, skeleton, retroperitoneum and left orbit. The diagnosis was based on the symmetric osteosclerosis of the long bones diaphyses (tibias and fibulas), breast lump histopathological/immunohistochemical findings and retroperitoneum and left orbit images in magnetic resonance. Mammary involvement by ECD is an extremely rare condition, which should be differentiated from some benign and malignant mimickers, especially the histiocytoid type of breast carcinoma. Characteristic histological features plus clinical and radiographic information are needed to achieve a correct diagnosis. The ECD, its relation to the LCH and details of the breast lesion are discussed. PMID- 15338305 TI - Expression of annexin II in conventional renal cell carcinoma is correlated with Fuhrman grade and clinical outcome. AB - Conventional renal cell carcinomas (CRCCs) were investigated for the expression of annexin II (ANX II) to determine out whether this calcium-binding protein could serve as a useful prognostic marker. CRCCs and adjacent nonneoplastic tissue from 33 patients were investigated for ANX II by immunohistochemistry, RT PCR, and western blot analysis. ANX II expression was correlated with tumor differentiation (Fuhrman grade) and to clinical outcome. Tumors were composed of ANX II positive and negative cells. In grade I tumors only a weak membranous staining was seen in immunopositive cells. In grade II and III tumors, however, ANX II was seen in the cytoplasm and at the cell membranes of tumor cells. On serial sections membranous and cytoplasmic immunoreactivity for ANX II occurred predominantly in eosinophilic cells whereas clear cells were mostly immunonegative. The ANX II expression in CRCCs was correlated with clinical outcome and Fuhrman grade. Since ANX II expression is correlated with Fuhrman grade and clinical outcome it may be a useful marker for prognosis in CRCC. PMID- 15338306 TI - Analysis of ascidian Not genes highlights their evolutionarily conserved and derived features of structure and expression in development. AB - The ascidian larva is often regarded as an organism close to the ancestral form of chordates, while it is generally accepted that the Spemann's organizer is absent from ascidian embryos. Not is one of the genes expressed in the organizer to execute functions in vertebrate embryos. To address the extent of conservation of Not gene expression among ascidians and vertebrates, we examined the structure and developmental expression of Not of the two distantly related ascidian species, Halocynthia and Ciona. Putative ascidian Not proteins were noted by the absence of one of the two motifs conserved among Not proteins of sea urchin and vertebrates. Analysis by in situ hybridization revealed that Not gene expression of ascidians could be categorized into three types: expression likely to be conserved between ascidians and vertebrates, that probably unique to ascidians, and that specific to ascidian species. Expression of ascidian Not in the posterior end of the tail as well as the notochord and a small part of the anterior neural tube at the tailbud stage is reminiscent of the expression of the vertebrate counterparts in the tailbud, which is regarded as a continuation of the organizer and the pineal gland, respectively. The expression of Not in the epidermis precursors during cleavage stage may be unique to ascidians. In the light of the present findings, evolutionary aspects of Not genes are discussed. PMID- 15338307 TI - Studies on water transport through the sweet cherry fruit surface: IX. Comparing permeability in water uptake and transpiration. AB - Water uptake and transpiration were studied through the surface of intact sweet cherry (Prunus avium L.) fruit, exocarp segments (ES) and cuticular membranes (CM) excised from the cheek of sweet cherry fruit and astomatous CM isolated from Schefflera arboricola (Hayata) Hayata, Citrus aurantium L., and Stephanotis floribunda Brongn. leaves or from Lycopersicon esculentum Mill. and Capsicum annuum L. var. annuum Fasciculatum Group fruit. ES and CM were mounted in diffusion cells. Water (deionized) uptake into intact sweet cherry fruit, through ES or CM interfacing water as a donor and a polyethyleneglycol (PEG 6000, osmotic pressure 2.83 MPa)-containing receiver was determined gravimetrically. Transpiration was quantified by monitoring weight loss of a PEG 6000-containing donor (2.83 MPa) against dry silica as a receiver. The permeability coefficients for osmotic water uptake and transpiration were calculated from the amount of water taken up or transpired per unit surface area and time, and the driving force for transport. Permeability during osmotic water uptake was markedly higher than during transpiration in intact sweet cherry fruit (40.2-fold), excised ES of sweet cherry fruit (12.5- to 53.7-fold) and isolated astomatous fruit and leaf CM of a range of species (on average 23.0-fold). Partitioning water transport into stomatal and cuticular components revealed that permeability of the sweet cherry fruit cuticle for water uptake was 11.9-fold higher and that of stomata 56.8-fold higher than the respective permeability during transpiration. Increasing water vapor activity in the receiver from 0 to 1 increased permeability during transpiration across isolated sweet cherry fruit CM about 2.1-fold. Permeability for vapor uptake from saturated water vapor into a PEG 6000 receiver solution was markedly lower than from liquid water, but of similar magnitude to the permeability during self-diffusion of (3)H(2)O in the absence of osmotica. The energy of activation for self-diffusion of water across ES or CM was higher than for osmotic water uptake and decreased with increasing stomatal density. The data indicate that viscous flow along an aqueous continuum across the sweet cherry fruit exocarp and across the astomatous CM of selected species accounted for the higher permeability during water uptake as compared to self-diffusion or transpiration. PMID- 15338308 TI - Reversal or reduction of glutamate and GABA transport in CNS pathology and therapy. AB - A dysfunction of amino acid neurotransmitter transporters occurs in a number of central nervous system disorders, including stroke, epilepsy, cerebral palsy and amyotrophic lateral sclerosis. This dysfunction can comprise a reversal of transport direction, leading to the release of neurotransmitter into the extracellular space, or an alteration in transporter expression level. This review analyses the role of glutamate and GABA transporters in the pathogenesis and therapy of a number of acute and chronic neurological disorders. PMID- 15338309 TI - The steroid RU486 induces UCP1 expression in brown adipocytes. AB - RU486 (mifepristone), a potent antagonist at progesterone and glucocorticoid receptors (PR and GR), is well known for its use in the termination of unwanted pregnancies, the potential development of oral contraceptives, treatment of certain cancers and other activities. Potentially, it could also play a role in obesity control, although the few studies that have addressed this aspect have focused mainly on its central and anti-glucocorticoid effects. We have shown previously that it could have a direct effect on brown adipocytes in culture when administered together with progesterone. The aim of the present work was to analyse the effects of RU486 on the expression of uncoupling proteins (UCPs) in brown adipocytes. In culture-grown, differentiated brown adipocytes, placed in a serum-free medium to exclude the presence of progesterone or glucocorticoids, RU486 stimulated UCP1 expression at both the mRNA and protein levels. These effects could be mediated by PR, GR or other unknown mechanisms but do not seem to be due to its anti-progestin or anti-glucocorticoid actions. The results suggest that the steroid RU486 has a direct action on adipocytes which could be useful for stimulating non-shivering BAT thermogenesis and therefore is of interest in obesity studies. PMID- 15338310 TI - Safe closure technique for distal pancreatic resection. AB - BACKGROUND AND AIMS: Various methods had previously been employed to manage the proximal pancreas after distal resection (mattress sutures with duct ligation; pancreato-enterostomy or stapling with stainless steel staples, etc.), with postoperative complications in 13% (6%-30%) of the cases, on average. In our practice, to reduce these complications, we applied staples made from Polysorb (Auto Suture), an absorbable lactomer. PATIENTS/METHODS: In the past 10 years, distal pancreatic resection in 90 patients [62 men, 28 women, mean age 52 (24 72)] years) was followed by closure of the resection surfaces with absorbable lactomer clips. Indications for distal resection (with or without splenectomy) were: focal pancreatic necrosis, spontaneous pancreatic fistulas, abscess, pseudocyst, traumatic disruption, segmental chronic obstructive pancreatitis in the tail, and benign (cystadenoma, or insulinoma) or malignant tumours. RESULTS: The postoperative period was uneventful in all these patients, without any complications (pancreatic fistula, abscess or bleeding). No morbidity or mortality occurred in the follow-up period (6 or 12 months postoperatively) with the exception of one patient who suffered a pseudocyst 6 months after surgery and was treated by cysto-jejunostomy. CONCLUSIONS: The clinical results clearly demonstrated that the application of absorbable lactomer staples for closure of the transected margin of the pancreas is a safe alternative to the standard closure technique. These staples can be applied in all cases when distal pancreatic resection is indicated for benign or malignant disorders or a traumatically injured pancreatic gland. PMID- 15338312 TI - Salivary analyses and caries increment over 4 years: an approach by cluster analysis. AB - MAIN PROBLEM: Caries as a multifactorial process is influenced by salivary defense. Cluster analyses should give additional information on the role of salivary variables in relation to caries increment. METHODS: Samples of unstimulated and stimulated whole saliva from 28 young adults (mean age 23.5+/ 2.1 years) were analyzed for flow rate, pH and buffer variables, lysozyme, lactoferrin, peroxidase, thiocyanate, secretory immunoglobulin A, and total protein. The decayed, missing, and filled surfaces (DMFS) were recorded at baseline and after 4 years. Cluster analyses were executed on the basis of salivary data. RESULTS: The mean caries increment (DeltaDMFS) over 4 years was 6.7+/-4.0 (range 1-16). In two-cluster processing, three out of four volunteers with low caries increments were grouped into one cluster. Only a few variables proved to be important for cluster characteristics. CONCLUSIONS: The results suggest that over 4 years (1) the volunteers with very low caries increment (DeltaDMFS=1) are classified always together, (2) these volunteers do not form a separate cluster by themselves, (3) low caries increment was related to higher salivary flow rate and lower levels of lysozyme and lactoferrin for unstimulated saliva and (4) the partial pressure of CO(2) was of importance in stimulated saliva. PMID- 15338314 TI - 153Sm and 166Ho complexes with tetraaza macrocycles containing pyridine and methylcarboxylate or methylphosphonate pendant arms. AB - A set of tetraaza macrocycles containing pyridine and methylcarboxylate (ac(3)py14) or methylphosphonate (MeP(2)py14 and P(3)py14) pendant arms were prepared and their stability constants with La(3+), Sm(3+), Gd(3+) and Ho(3+) determined by potentiometry at 25 degrees C and 0.10 M ionic strength in NMe(4)NO(3). The metal:ligand ratio for (153)Sm and (166)Ho and for ac(3)py14, MeP(2)py14 and P(3)py14, as well as the pH of the reaction mixtures, were optimized to achieve a chelation efficiency higher than 98%. These radiocomplexes are hydrophilic and have a significant plasmatic protein binding. In vitro stability was studied in physiological solutions and in human serum. All complexes are stable in saline and PBS, but 20% of radiochemical impurities were detected after 24 h of incubation in serum. Biodistribution studies in mice indicated a slow rate of clearance from blood and muscle, a high and rapid liver uptake and a very slow rate of total radioactivity excretion. Some bone uptake was observed for complexes with MeP(2)py14 and P(3)py14, which was enhanced with time and the number of methylphosphonate groups. This biological profile supports the in vitro instability found in serum and is consistent with the thermodynamic stability constants found for these complexes. PMID- 15338315 TI - Relapse of depression during pregnancy following antidepressant discontinuation: a preliminary prospective study. AB - OBJECTIVE: Pregnancy has frequently been referred to as a time of emotional well being for patients. However, systematic data about the risk for relapse of depression during pregnancy are sparse. METHOD: We completed a longitudinal cohort study of thirty-two (N = 32) women with histories of depression who were euthymic at conception and who either discontinued or attempted to discontinue antidepressant therapy proximate to conception. Subjects were prospectively followed across pregnancy once per trimester using structured clinical interviews. Rates of relapse and time to relapse were examined. Factors distinguishing the population with respect to risk for relapse including demographic characteristics and illness history were also examined. RESULTS: Seventy-five percent (N = 24) of patients relapsed during pregnancy. The majority of relapses (79%, N = 19) occurred in the first trimester, and relapse was more prevalent in women with histories of more chronic depression. CONCLUSIONS: Pregnancy is not "protective" with respect to risk for relapse of depression. Careful treatment planning is necessary for those women on antidepressants who plan to conceive or who become pregnant. PMID- 15338316 TI - Routine psychosocial assessment of women in the antenatal period: frequency of risk factors and implications for clinical services. AB - Routine psychosocial assessment was introduced at an Australian public hospital's antenatal clinic in 2001. After modification, this assessment consists of 12 questions together with the Edinburgh Depression Scale (EDS). Data are reported for responses to these questions from over 2,000 English-speaking women presenting to the clinic in a 12-month period. These 12 questions and the EDS were categorised into seven risk domains, with 12% of the women (n = 260) having three or more of these risk domains. Referral information to one of our two clinical services shows that 6.7% of women assessed in the antenatal clinic become clients having face-face counselling, and a further 7.2% have just telephone contact with this specialist perinatal mental health service. This information should prove useful for services considering implementing routine psychosocial assessments (or "screening") in the antenatal period. PMID- 15338317 TI - Betaine or taurine administration prevents fibrosis and lipid peroxidation induced by rat liver by ethanol plus carbon tetrachloride intoxication. AB - The aim of this study was to investigate the effect of betaine or taurine on liver fibrogenesis and lipid peroxidation in rats. Fibrosis was induced by treatment of rats with drinking water containing 5% ethanol and CCl(4) (2 x weekly, 0.2 ml/kg, i.p.) for 4 weeks. Ethanol plus CCl(4) treatment caused increased lipid peroxidation and disturbed antioxidant system in the liver. Histopathological findings suggested that the development of liver fibrosis was prevented in rats treated with betaine or taurine (1% v/v in drinking water) together with ethanol plus CCl(4) for 4 weeks. When hepatic taurine content was depleted with beta-alanine (3% v/v in drinking water), portal-central fibrosis induced by ethanol + CCl(4) treatment was observed to proceed cirrhotic structure. Betaine or taurine was also found to decrease serum transaminase activities and hepatic lipid peroxidation without any change in hepatic antioxidant system in rats with hepatic fibrosis. In conclusion, the administration of betaine or taurine prevented the development of liver fibrosis probably associated with decreased oxidative stress. PMID- 15338318 TI - Interferon type I response in porcine reproductive and respiratory syndrome virus infected MARC-145 cells. AB - Infection by porcine reproductive and respiratory syndrome virus (PRRSV) results in a weak induction of the innate immune response. There are many genes that collectively comprise this response and the extent to which each gene responds to PRRSV infection is unclear and warrants further investigation. To this end, we have utilized real-time PCR using SYBR Green I dye-based detection to quantify transcript abundance of the type I interferons (IFN-alpha and -beta) and IFN-beta transcriptional enhanceasome genes. In MARC-145 cells, both IFN-alpha and -beta transcript abundance were unaffected by PRRSV infection. However, stimulation of MARC-145 cells by exogenous double-stranded RNA, resulted in significant increases in transcript abundance of both IFN-alpha and -beta as well as IFN-beta enhanceasome components, indicating that a type I IFN response could be induced in these cells. The double-stranded RNA induction of type I IFN transcription was significantly inhibited by dual-exposure with PRRSV. These results suggest that PRRSV infection directly interferes with type I IFN transcriptional activation early in its pathway, at the level of IFN-beta gene transcription. PMID- 15338319 TI - Passive protection to bovine rotavirus (BRV) infection induced by a BRV VP8* produced in plants using a TMV-based vector. AB - We have previously reported on the use of a tobacco mosaic virus (TMV) vector TMV 30B to express foreign viral antigens for use as experimental immunogens. Here we describe the development of an improved TMV-30B vector that adds a sequence of 7 histidine residues to the C-terminus of recombinant proteins expressed in the vector. We used this TMV-30B-HISc vector to express the VP8* fragment of the VP4 protein from bovine rotavirus (BRV) strain C-486 in plants. Recombinant VP8* protein was purified from N. benthamiana leaves at 7 days post-inoculation by immobilized metal affinity chromatography. The plant-produced VP8* was initially detected using anti-His tag mAb and its antigenic nature was confirmed using both monoclonal and polyclonal specific antisera directed against BRV. Adult female mice, inoculated by the intraperinoteal route with an immunogen containing 4 microg of recombinant VP8*, developed a specific and sustained response to the native VP8* from the homologous BRV. Eighty five percent of suckling mice from immunized dams that were challenged with the homologous virus at the fifth day of age were protected from virus as compared to 35% of the pups from mothers immunized with a control protein. These results demonstrate that the plant produced VP8* was able to induce passive protection in the new born through the immunization of dams. This suggests that the technology presented here provides a simple method for using plants as an inexpensive alternative source for production of recombinant anti-rotavirus antigens. PMID- 15338320 TI - LCMV-mediated hepatitis in rhesus macaques: WE but not ARM strain activates hepatocytes and induces liver regeneration. AB - Lymphocytic chorimeningitis virus (LCMV), the prototype arenavirus, and Lassa virus (LASV), causative agent of Lassa hemorrhagic fever (LHF), belong to the Old World group of the family Arenaviridae. Both viruses have extensive strain diversity and significant variations in lethality and pathogenicity for man and experimental animals. We have shown that the LHF-like infection of rhesus macaques with the WE strain of LCMV affects liver functions, induces hepatocyte proliferation, and causes a rise in IL-6 and soluble TNF receptors (sTNFR) concomitant with a rise in viremia. The levels of IL-6 and sTNFR can serve as an additional diagnostic tool for liver involvement in pathogenesis of arenavirus infection. Mucosal inoculation of rhesus macaques with LCMV-WE can result in attenuated infection with a transient viremia and liver enzyme abnormalities. The ARM strain of LCMV shares 88% amino acid homology with WE. In contrast to LCMV WE, ARM strain does not induce manifested disease in monkeys, does not affect liver functions, and does not induce hepatocyte proliferation. Previously we demonstrated that LCMV-ARM infection protected rhesus macaques challenged with LCMV-WE. Here we have shown that the protected animals have no signs of hepatitis and hepatocyte proliferation. PMID- 15338321 TI - Mutational analysis of the HIV-1 LTR as a promoter of negative sense transcription. AB - The HIV-1 gene promoter is a bi-directional promoter of transcription. We report the characterization of the negative sense promoter (NSP) by analysis of the effect on negative sense transcription of a series of LTR U3 region substitution mutants. Mutations in the region nt -58 to -183 (positive sense transcription initiation nt +1) reduced transcription to <15% of wild type NSP activity. This region, essential for NSP activity, was designated the core basal promoter. Over expression of NF-kappaB RelA(p65) and LEF-1 increased negative sense expression, as did over expression of H-ras oncogene, consistent with the presence of cognate sequence motifs for NF-kappaB, LEF-1 and RBF. We were also able to confirm that the NSP is a TATA-less promoter inhibited by HIV-1 Tat. Based on our findings, we propose a model for the interaction between the NSP and PSP, and the role of Tat in regulating the interaction. PMID- 15338322 TI - In vitro cytoskeleton changes of mouse neurons induced by purified HTLV-1, and PBMC from HAM/TSP patients and HTLV-1 carriers. AB - HTLV-1 is the causative agent of HAM/TSP. This neurological disease affects the CNS producing damage of the motor tracts at the spinal cord. The HAM/TSP pathogenesis remains undefined. It could include direct and indirect actions of HTLV-1. We studied the effect of purified HTLV-1 and the PBMC of 22 Chilean patients co-cultivated with fetal neurons of mouse (CNh cells): 8 HAM/TSP, 8 HTLV 1 carriers, and 6 non-infected controls. The viral antigens and provirus in CNh cells was evaluated with monoclonal and polyclonal antibodies reacting with HTLV 1 by immunofluorescence assay and PCR at 0, 7 and 15 days of co-cultures, respectively. Viral antigens were detected in 0.1-0.5%, and 0-0.3% of the neurons incubated with lymphocytes of HAM/TSP patients and HTLV-1 carriers, respectively. Neurons incubated with cells of 7 HAM/TSP patients, and 3 HTLV-1 carriers showed the presence of nucleotide sequences of tax gene. These results would be showing that CNh cells would express viral antigens and provirus. The HTLV-1 or their proteins were capable in vitro to produce structural and growth changes in the cytoskeleton of CNh neurons. In this series, the purified HTLV-1 was more effective in the neural changes than PBMC of HAM/TSP or HTLV-1 carriers. PMID- 15338323 TI - A derivative of bovine herpesvirus 1 (BoHV-1) UL3.5 lacking the last forty amino acids inhibits replication of BoHV-1. AB - Bovine herpesvirus 1 (BoHV-1) UL3.5 is a tegument protein that interacts with BoHV-1 alpha-transinducing factor alphaBTIF). In this report, we show that a truncated UL3.5 lacking the last 40 amino acids (C40UL3.5) inhibited replication of BoHV-1. Stable expression of C40UL3.5 in MDBK cells inhibited replication of BoHV-1 300- to 500-fold in plaque assays. This inhibitory effect was specific for BoHV-1 as cells expressing C40UL3.5 supported replication of pseudorabies virus (species Suid herpesvirus 1, SuHV-1) and herpes simplex virus (species Human herpesvirus 1, HHV-1) to normal levels. In contrast, a truncated UL3.5 which lacked the first 20 amino acids and did not interact with alphaBTIF did not inhibit BoHV-1 replication. In the C40UL3.5-expressing cells infected with BoHV 1, expression of the viral immediate early gene BICP4 and the early protein gB were reduced and delayed. C40UL3.5, when either transiently or stably expressed, inhibited alphaBTIF-mediated transactivation of a BoHV-1 immediate-early promoter. C40UL3.5 may be useful for constructing transgenic cattle resistant to infection by BoHV-1. PMID- 15338324 TI - Herpes simplex virus types 1 and 2 infect the mouse pituitary gland and induce apoptotic cell death. AB - Herpes simplex virus (HSV) infected the anterior lobe of the pituitary gland resulting in cytopathic changes following intravenous (i.v.) inoculation of male mice. Both HSV type 1 (HSV-1) and type 2 (HSV-2) were isolated from pituitary gland following i.v. infection, but not after intraperitoneal inoculation. HSV infected pituitary cells were microscopically visible beginning at 24 h or 48 h following i.v. inoculation and were localized in the anterior pituitary. In both HSV-1 and -2 infections the pituitary lesions were apoptotic, as determined by light and electron microscopy, TUNEL, and DNA gel electrophoresis. However, the pituitary infection does not appear to be life-threatening since pituitary lesions were also observed following i.v. infection with HSV-1 strain -GC which possesses low virulence. These results suggest that the pituitary gland is one of the target organs of HSV infection. PMID- 15338325 TI - Variability in the coat protein gene of Papaya ringspot virus isolates from multiple locations in India. AB - The coat protein (CP) sequences of eleven Papaya ringspot virus (PRSV) isolates originating from different locations in India were determined, analysed and compared with the sequences of other isolates of PRSV. The virus isolates from India exhibited considerable heterogeneity in the CP sequences. The CP-coding region varied in size from 840-858 nucleotides, encoding protein of 280-286 amino acids. Comparative sequence analysis revealed that the PRSV isolates originating from India were divergent up to 11%. Though the PRSV isolates were differentiated in to two clusters, yet the sequence variation could not be correlated with the geographical origin of the isolates. Implication of the sequence variation in the coat protein derived transgenic resistance in papaya is discussed. PMID- 15338326 TI - Epitope analysis of monoclonal antibody E5/G6, which binds to a linear epitope in the nucleocapsid protein of hantaviruses. AB - Monoclonal antibody E5/G6 recognized a linear epitope common to hantavirus nucleocapsid proteins. Using synthetic peptides, we identified epitope E5/G6 as the 9 mer YEDVNGIRK (NP 165-173), in which D167, G170, I171, and R172 are indispensable. Furthermore, all the peptides synthesized using various hantavirus sequences bound MAb E5/G6 consistently, despite the existence of several amino acid variations in this region. These results indicate that MAb E5/G6 is a useful tool for detecting hantavirus antigen in rodent or patient tissues using Western blotting or other immunohistochemical assays. PMID- 15338327 TI - Molecular variability analysis of five new complete cacao swollen shoot virus genomic sequences. AB - Cacao swollen shoot virus (CSSV), a member of the family Caulimovi-ridae, genus Badnavirus occurs in all the main cacao-growing areas of West Africa. We amplified, cloned and sequenced complete genomes of five new isolates, two originating from Togo and three originating from Ghana. The genome of these five newly sequenced isolates all contain the five putative open reading frames I, II, III, X and Y described for the first sequenced CSSV isolate, Agou1 originating from Togo. Their genomes have been aligned with the genome of Agou1. The nucleotide and amino acid sequence identities between isolates have been calculated and a phylogenetic analysis has been made including other pararetroviruses. Maximum nucleotide sequence variability between complete genomes of CSSV isolates was 29.4%. Geographical differentiation between isolates appears more important than differentiation between mild and severe isolates. ORF X differs greatly in size and sequence between the Togolese isolates Nyongbo2 and Agou1, and the four other isolates, its functional role is therefore clearly questionable. PMID- 15338328 TI - Pitavastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, reduces hippocampal damage after transient cerebral ischemia in gerbils. AB - Pitavastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, is a potent cholesterol-lowering drug that reduces the risk of myocardial infarction and stoke. In this study, we examined its neuroprotective effects against hippocampal CA1 neuronal damage following transient cerebral ischemia in gerbils. Forebrain ischemia was induced by occlusion of bilateral common carotid arteries for 5 min. Pitavastatin, at a dose of 3, 10 or 30 mg/kg, was administered orally twice a day for 5 consecutive days and transient cerebral ischemia was induced in mice 1 h after the last treatment with pitavastatin. Histopathological observations showed that neuronal damage to the hippocampal CA1 neurons, which was observed 5 days after ischemia in animals, was prevented by pitavastatin treatment. Immunohistochemical staining for copper/zinc superoxide dismutase (SOD) and manganese SOD decreased in the hippocampal CA1 sector of gerbils 2 days after ischemia when histological neuronal destruction was not yet found, but was clearly observed in pitavastatin-treated animals. These results indicate that pitavastatin can protect dose-dependently against ischemia-induced neuronal damage and that the mechanism of the neuroprotection may be related to the preservation of SODs, especially copper/zinc-SOD. This in part explains how pitavastatin therapy, which targets free radicals, has beneficial effects against disorders including ischemic stroke. PMID- 15338329 TI - Effects of piracetam alone and in combination with antiepileptic drugs in rodent seizure models. AB - The nootropic drug piracetam was investigated in various experimental models of epilepsy. Generally, piracetam exhibits no or only moderate anticonvulsant properties against generalized tonic or clonic seizures. However, in many cases it did increase the anticonvulsant effectiveness of conventional antiepileptics, as shown in the maximal electroshock seizure (MES) threshold test, the traditional MES test or in DBA/2 mice. A pharmacokinetic interaction does not seem to be responsible for this effect. In lethargic mice, a model of absence seizures, piracetam significantly decreased the incidence and duration of spike wave discharges. Furthermore, in the cobalt-induced focal epilepsy model piracetam reduced the number of spikes/min and in the hippocampal stimulation model it increased the anticonvulsant potency of phenobarbital and phenytoin after single and repeated administration. In conclusion, the well tolerated piracetam itself did not show marked anticonvulsant effects in most screening tests, however, its co-medication with antiepileptic drugs improved seizure protection in various models which may bear potential clinical significance. PMID- 15338330 TI - Aging and working memory: early deficits in EEG activation of posterior cortical areas. AB - Using the n-back task, we recently identified, in young subjects, a positive negative event related potential component (PN(wm)) in a time-range window between 140 and 280 ms after stimulus onset representing an electrophysiological correlate of working memory load. To evaluate age-related electrophysiological changes in working memory processing, we applied the same neuropsychological paradigm and compared densities of the PN(wm) component in 17 young (mean age: 26) and 17 healthy elderly individuals (mean age: 75). Both age groups displayed a PN(wm) component during the two working memory tasks. For the 1-back task, densities were similar in both young and elderly individuals. In contrast, PN(wm) densities increased with higher memory load (2-back>1-back) in the younger but not in the older group. This difference was mainly observed over parietal electrodes suggesting an impaired activation of neural generators within this brain region. The present results are consistent with the hypothesis of decreased brain reserve in the elderly and provide evidence for age-related deficits in the recruitment of posterior cortical neurons with increasing working memory load. PMID- 15338331 TI - Mitochondrial genotype and risk for Alzheimer's disease: cross-sectional data from the Vienna-Transdanube-Aging "VITA" study. AB - The Vienna Transdanube Aging (VITA) study searches for early markers of Alzheimer's disease (AD) by examining the mental status in a community-based cohort of 606, 75-years old volunteers that are then related to various clinical and genetic analyses. To determine whether mutations in mtDNA are involved in expression of AD, the mtDNA of 79 "control" participants is screened for alterations by sequencing of "hot-spot-regions". This study on mtDNA mutations has eliminated the influence of aging on the occurrence of mtDNA alterations by sequencing samples from persons at the age of exactly 75 years. Thus, our cohort reveals a snap-shot of mitochondrial sequences of elderly persons. So far, a high percentage (56%) of persons with known or unknown mutations in the fragments analyzed were found. These data will be compared in due time to a cohort of participants with proven late-onset AD. PMID- 15338332 TI - Impact of gender on upregulation of antioxidant defence mechanisms in Alzheimer's disease brain. AB - Since oxidative stress plays an important role in the pathogenesis of Alzheimer's disease (AD) and since the age-adjusted incidence of AD is higher in females than males, we examined a possible influence of gender on antioxidant metabolism in brains from male and female AD patients and age-matched controls. Activities of copper/zinc-dependent superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-disulfide reductase (GR) were elevated in AD samples compared to controls. Upon in vitro stimulation, levels of malondialdehyde formation were significantly lower in AD samples, probably due to the increased antioxidant capacity. Overall, our results indicate that antioxidant metabolism is functionally still intact but increased in AD implying that oxidative damage is caused rather by overproduction than by insufficient detoxification of ROS. Among AD patients, a gender-specific partial upregulation of antioxidant defence was present: activities of SOD and GPx were even further increased in female patients, and levels of 4-hydroxynonenal, a marker of oxidative damage, were higher than in male patients. Importantly, our results are in line with epidemiological studies indicating a higher risk for AD in females. Thus, gender differences in oxidative stress parameters might be related to the higher prevalence of AD in females. PMID- 15338333 TI - Interleukin-1 gene cluster polymorphisms and risk of Alzheimer's disease in Chinese Han population. AB - Interleukin-1 (IL-1) has been implicated as a key cytokine in Alzheimer's disease (AD) pathogenesis. IL-1 gene polymorphisms, especially IL-1A C((-)889)T polymorphism, have been suggested to be associated with AD risk and onset age. To determine if IL-1 polymorphisms are genetic risk factors for developing AD in Chinese Mainland population, we analyzed IL-1A ((-)889), IL-1B ((-)511) and IL 1RN variable number of tandem repeat (VNTR) polymorphisms in a sample of 145 sporadic AD patients and 181 healthy controls. Our data revealed that the three polymorphisms in IL-1 gene cluster might not play a key role in AD pathogenesis in Chinese Mainland Han population. PMID- 15338334 TI - Cu, Zn- and Mn-superoxide dismutase levels in brains of patients with schizophrenic psychosis. AB - Impaired oxidative stress defense has been reported in blood of both drug-naive and antipsychotic-treated patients suffering from schizophrenic psychosis, indicating the involvement of free radical metabolism in the pathogenetic processes of schizophrenia. In this study, the concentrations of two isoenzymes of superoxide dismutase (SOD), Cu, Zn- and MnSOD, were determined with ELISA in various cortical (frontal, parietal, temporal and occipital cortex) and subcortical areas (putamen, caudate nucleus, thalamus, and substantia innominata) of post-mortem brain tissue from patients diagnosed with a schizophrenia spectrum disorder and compared with those of controls. Post-mortem brain tissue from individuals without neuropsychiatric disorders served for control. Cu, Zn- and MnSOD levels were significantly increased in frontal cortex and substantia innominata of the index group, respectively. In all other areas both types of SOD remained virtually unchanged. Detection of SOD changes in the brain supports previous reports of alterations of antioxidant indices in blood cells of patients with schizophrenia and suggests a specific neuroanatomical distribution pattern of oxidative stress processes possibly related to the pathophysiology of schizophrenia. PMID- 15338335 TI - Bench test assessment of the new Raumedic Neurovent-P ICP sensor: a technical report by the BrainIT group. AB - BACKGROUND: In clinical practice, fiberberoptic and piezo-electric ICP probes are often used for measuring intracranial pressure (ICP). A number of similar technologies, although performing well in bench test studies, have been shown to exhibit unacceptable zero drift, fragility or both during trials conducted under clinical conditions. Recently, a new technology has become available, the Neurovent-P (Raumedic AG + CO, Raumedic, Germany). As a pre-requisite for a clinical trial, we have conducted and report on bench test studies to confirm the manufacturer's long term zero-drift performance for this technology. METHOD: In a test rig static tests (recording of 20 mmHg pressure) and dynamic tests, ranging from 5 to 50 mmHg have been performed. FINDINGS: 10 ICP probes have been tested for a total of 60 days. All the catheters, after the connection with the ICU monitor displayed a static pressure of 0 +/- 1 mmHg and did not required pre insertion alteration. At five days, mean zero drift was 0.6 +/- 0.9 mmHg. Overall, zero drift ranged from 0 to 2 mmHg. At a fixed static pressure of 20 mmHg, the mean recorded value was 20.6 +/- 0.8 mmHg, ranging from 19 to 23 mmHg. A regression analysis of the relationship between the applied pressure and the recorded pressure during the dynamic tests of the 10 catheters yielded a correlation coefficient R2 of 0.997. Applying the Altman and Bland method to assess the bias and confidence limits for the Raumedic catheter responses during the dynamic tests against the applied gold-standard hydrostatic column pressures, the average bias of -0.66 +/- 0.85 mmHg, with 95% CLs of -2 mmHg and 1 mmHg. CONCLUSIONS: Mean zero drift, after five days, was very small and long-term continuous recording of a stable pressure was very precise. The response at dynamic tests, i.e. the changes of pressure in a wide range, was excellent. The average bias of the Raumedic catheter compared with the hydrostatic column is very small. After this bench test, the next and most critical step will be to conduct a trial of this promising technology under more demanding clinical environment. PMID- 15338336 TI - Neurosurgical interventions during pregnancy and the puerperium: clinical considerations and management. AB - Neurosurgical interventions during pregnancy represent a special clinical challenge and require a meticulously selected treatment strategy. A thorough understanding of the underlying pathophysiological mechanisms involving mother and fetus is mandatory. We report our experience with neurosurgical procedures during pregnancy and the puerperium and summarize the difficulties encountered. The emerging diagnostic as well as therapeutic implications in these complex cases are also discussed. PMID- 15338337 TI - Spondylodiscitis caused by occult endocarditis. PMID- 15338338 TI - Successful resection of a left insular cavernous angioma using neuronavigation and intraoperative language mapping. AB - Despite recent literature advocating the surgical removal of symptomatic Cavernous Angiomas (CA), even in critical brain areas, very few observations of insular CA surgery have been described, particularly in the left hemisphere. We report the case of a successful resection of a CA located in the dominant insula, using both neuronavigation and intra-operative functional mapping. This 33-year old right-handed man harbored a left insular CA, revealed by generalized seizures following a bleed confirmed on MRI. The preoperative examination was normal. A stereotactic-guided surgery was performed under local anesthesia, with intra operative functional mapping using direct cortico-subcortical electrical stimulation in the awake patient--allowing the surgeon to achieve total resection of both CA and pericavernomatous gliosis, as shown on repeated postoperative MRIs. There was no postsurgical deficit, nor any seizure without treatment (follow-up: 4.5 years). The diagnosis of CA was confirmed by histological examination. Taking account of the risk of morbidity due to the natural history of CA, particularly in eloquent brain regions, we suggest to routinely consider the possibility of a surgical treatment in cases of symptomatic (left dominant) insular CA, using combined intra-operative anatomical and physiological localization methods. PMID- 15338339 TI - Resolution of spinal epidural haematoma without surgery in a haemophilic infant. AB - Non-traumatic spinal epidural haematoma is a rare complication of haemophilia. We report a seven-month-old boy who presented with symptomatic spinal epidural haematoma. He was found to have a hemophilia B trait, and after factor IX replacement, his neurological signs were stabilized and follow-up MRI demonstrated rapid resolution of the haematoma. This illustrative case suggests that surgical intervention can be deferred as the first step in treating spinal epidural haematoma in a haemophilic infant. PMID- 15338340 TI - Intramedullary spinal cord abscess in a 4-year old child. PMID- 15338341 TI - Cauda-filar paraganglioma with 'silk cocoon' appearance on spinal angiography. AB - 'Silk cocoon' appearance on spinal angiography is pathognomonic to differentiate paragangliomas from several vascular tumors and malformations of cauda-filar region. PMID- 15338342 TI - Recovery of the susceptibility of isolated bacterium achieved by giving long established antibiotics as prophylaxis against postoperative infections. AB - PURPOSE: To evaluate the effectiveness of operative antibiotic therapy we changed the antibiotics and examined the susceptibility of the postoperative infection. METHODS: We studied 4 593 patients who underwent gastrointestinal surgery during the last 12.5 years, and examined the changes in intraoperative antibiotics, the return of bacteria isolated from infectious sites, the trends in frequency of Methicillin-resistant Staphylococcus aureus (MRSA) isolation, and changes in the antibiotic susceptibility of Pseudomonas aeruginosa and Bacterioides fragilis. We changed the antibiotics to Cefazolin (CEZ) for upper gastrointestinal tract surgery and cholecystectomy, and to Cefotiam (CTM) for colonic, liver, and pancreatic surgery. We also reduced the period of drug administration. RESULTS: The rate of MRSA infections decreased, the rate of P. aeruginosa infections was always under 20% and the rate of B. fragilis infections increased. After the guidelines of drug susceptibility were prepared, the minimum inhibitory concentrations (MICs) of Cefsulodin (CFS) and Piperacillin (PIPC) for P. aeruginosa and of Latamoxef (LMOX) and PIPC for B. fragilis, decreased. CONCLUSION: The use of antibiotics considered comparatively old for the prophylaxis of postoperative infection prevented the emergence of MRSA-like resistant strains, influenced changes in Gram-negative bacteria drug susceptibility, and led to an overall reduction in multiple drug resistance. PMID- 15338343 TI - Surgical management of substernal goiters: clinical experience of 170 cases. AB - PURPOSE: To discuss the presentation, diagnosis, treatment, histopathological findings, and complications of patients who underwent thyroidectomy for substernal goiter in our surgical clinic. METHODS: We retrospectively analyzed 170 patients with substernal goiters among 2650 patients undergoing surgical treatment for various thyroid diseases between 1990 and 2003. We evaluated the clinical data, preoperative diagnostic findings, surgical treatments, histopathological results, and postoperative complications. RESULTS: The most common symptoms were a cervical mass (88%) and dyspnea (35%), but 26% of the patients were asymptomatic. Chest radiography provided the first evidence of a substernal goiter in 77% of the patients. We performed total or near total thyroidectomy and operated through a cervical incision in all but 12 of the patients. There was no operative mortality but 12 (7%) patients suffered temporary hypoparathyroidism and 4 suffered transient vocal cord paralysis (2%). Malignancy was diagnosed by histopathological examination in 22 (13%) patients. CONCLUSION: We think that the diagnosis of a substernal goiter is an indication for thyroidectomy, which is associated with very low postoperative morbidity. PMID- 15338344 TI - Minilaparotomy approach for the resection of laterally spreading tumors of the colon. AB - PURPOSE: We report our experience of using the minimally invasive minilaparotomy approach to resect colonic laterally spreading tumors (LSTs) that could not be removed by colonoscopic snare polypectomy. METHODS: We prospectively examined 17 patients who underwent a minilaparotomy, defined as an incision less than 7 cm long, between 1997 and 2001, for a collective 19 colonic LSTs. RESULTS: Complete en bloc resection of the LSTs was successfully performed in all 17 patients. The resections included colotomy and polypectomy in four patients (four LSTs), limited colectomy in seven patients (nine LSTs), and colectomy with regional lymph node dissection in six patients (six LSTs). There was no mortality or morbidity. The mean +/- standard deviation (SD) length of the minilaparotomy was 6.7 (+/-0.8) cm, and the mean (+/-SD) operating time and blood loss were 139 (+/ 39) min and 27 (+/-15) ml, respectively. Histology revealed 2 adenomas, 16 Tis carcinomas, and 1 T1 carcinoma. None of the patients had lymph node metastasis or positive resection margins. There have been no signs of tumor recurrence after a median follow-up period of 30.4 months. CONCLUSIONS: The minilaparotomy approach is appropriate for resecting LSTs that cannot be removed by colonoscopic snare polypectomy, and provides a minimally invasive alternative to conventional laparotomy. PMID- 15338345 TI - Outcome of laparoscopic rectopexy for complete rectal prolapse in patients older than 70 years versus younger patients. AB - PURPOSE: To evaluate the outcome of laparoscopic rectopexy for complete rectal prolapse in patients above 70 years of age, compared with that in younger patients. METHODS: Between October 1997 and September 2001, 14 consecutive patients with complete rectal prolapse underwent laparoscopic rectopexy. Nine patients were aged 70 years or older, and five were aged under 70 years. All of the patients were ambulant and well enough to tolerate surgery under general anesthesia. Each patient was monitored pre- and postoperatively, for fecal incontinence, constipation, recurrent prolapse, morbidity, and mortality. RESULTS: The median follow-up period was 34.5 (range 5-54) months. No significant differences were noted in the hospitalization, incidence of complications, recurrence rate, and functional outcome. CONCLUSION: The outcome of laparoscopic rectopexy in elderly patients is similar to that in younger patients. Therefore, advanced age alone should not be a contraindication to laparoscopic rectopexy. PMID- 15338346 TI - Relationships between tensile strength, ascorbic acid, hydroxyproline, and zinc levels of rabbit full-thickness incision wound healing. AB - PURPOSE: This work was carried out to follow up the healing of full-thickness incision wounds opened on the back skin of rabbits in order to gain insight into the periodical correlation among such factors as ascorbic acid, collagen (hydroxyproline), the zinc content, and tensile strength of wound tissue. The need to provide vitamin C or zinc supplements after such wound incisions is also discussed. METHODS: Full-thickness incision wounds and the ascorbic acid and hydroxyproline levels were measured in 24 rabbits by spectrophotometric methods on day 0 and on the 3rd, 5th, 7th, and 15th days after operation. The tensile strength was measured by a polygraph using a force displacement transducer. The zinc levels of the wounds were measured by atomic absorption spectrophotometry. RESULTS: The zinc and hydroxyproline levels reached the peak levels on the 5th day, but the tensile strength of wound increased sharply on the 7th day after wounding while the zinc levels did not change. These results indicated that in the first 7 days of wound healing, high levels of ascorbic acid, hydroxyproline, and zinc cumulation occurred in the wound tissue, and the tensile strength reached its highest level on the 15th day without any supplementation. CONCLUSION: The supplementation of zinc and/or ascorbic acid should therefore be given just at the beginning of the wounding period, especially if there is deficiency of these nutrients. PMID- 15338347 TI - Myxofibrosarcoma of the breast as an unusual variant of malignant fibrous histiocytoma: report of a case. AB - Soft tissue sarcomas of the breast account for less than 1% of all malignant breast tumors. The majority of these lesions have an epithelial component and are thus classified as "cystosarcoma phyllodes." All other types of sarcomas are categorized according to the existing histological soft tissue classification. It is difficult to determine the relative frequency of the different types of breast sarcoma because there are wide variations among the reported series. We report a case of myxofibrosarcoma, a variant of malignant fibrous histiocytoma, in a 58 year-old woman. PMID- 15338348 TI - Breast cancer with nephrotic syndrome: report of two cases. AB - We report two cases of women found to have breast cancers within a few months of being diagnosed with nephrotic syndrome. Case 1 was a 53-year-old Japanese woman in whom breast cancer was diagnosed 14 months after the onset of nephrotic syndrome. The histological diagnosis was invasive ductal carcinoma with no lymph node metastasis. We performed a modified radical mastectomy, after which the proteinuria and hypoproteinemia resolved almost completely, and the patient has been disease-free for 5 years since. Case 2 was a 61-year-old Japanese woman in whom breast cancer was diagnosed 2 months after the onset of membranous nephropathy. We performed a modified radical mastectomy and the histological diagnosis was invasive ductal carcinoma with marked lymphatic vessel permeation and involvement of five axillary lymph nodes. Proteinuria and hypoproteinemia did not resolve postoperatively and there is a high possibility of remnant or recurrent cancer. To our knowledge, there are only four other reported cases of paraneoplastic membranous nephropathy complicating breast cancer. However, we speculate that the postoperative resolution of nephrotic syndrome might be a measure of cancer control. PMID- 15338349 TI - Granular cell tumor of the breast preoperatively diagnosed by fine-needle aspiration cytology: report of a case. AB - Granular cell tumors (GCTs) have a characteristic cytological appearance, and fine-needle aspiration cytology (FNAC) has been suggested to be the diagnostic modality of choice. However, the differential diagnosis has not yet been well described. We herein describe a rare case of GCT of the breast and discuss the differential diagnosis. A 53-year-old woman presented to our hospital with a left breast mass. In clinical and radiological examinations, the mass was suspected to be malignant. Cytologically, the tumor was composed of cohesive groups of cells with a syncytial appearance, and the cells contained abundant, finely eosinophilic cytoplasmic granules and small round-to-slightly-oval nuclei, thus suggesting the presence of benign GCT. The results of immunohistochemical staining supported the proposed deviation from Schwann cells. This case emphasizes the fact that GCTs are a rare but important possibility in the differential diagnosis of breast tumors, and that FNAC may provide clinically useful information on the management of such lesions. PMID- 15338350 TI - Acinic cell carcinoma of the trachea: report of a case. AB - Acinic cell carcinoma (ACC) is a very rare primary tracheal carcinoma. We report a case of primary ACC of the trachea, which represents, to the best of our knowledge, only the fifth such case ever documented. Pharyngolaryngoesophagectomy was necessary for complete resection because the tumor invaded the trachea widely, up to the cricoid cartilage and involving the cervical esophagus. Although invasion of the upper aerodigestive tract by neoplasma is rare, to achieve complete resection and an acceptable prognosis, even a tumor with low malignancy requires wide resection. PMID- 15338351 TI - Neurilemoma originating from the left recurrent nerve in the superior mediastinum: report of a case. AB - We report a rare case of neurilemoma originating from the left recurrent nerve in the superior mediastinum. PMID- 15338352 TI - Posterior mediastinal localization of Castleman's disease: report of a case. AB - Castleman's disease, defined as angiofollicular lymphoid hyperplasia, is a rare lymphoproliferative disorder, which usually occurs in the chest. The tumor is often asymptomatic, but it can cause nonspecific thoracic symptoms such as cough and dyspnea. Surgical removal is curative and malignant transformation has not been described. We report an unusual case of Castleman's disease localized in the posterior mediastinum and bordering the chest wall, and review the relevant literature. PMID- 15338353 TI - Transaortic mitral valve repair combined with a modified Bentall procedure in a patient with Marfan's syndrome: report of a case. AB - We report a case of transaortic mitral valve repair combined with aortic root and arch replacement in a patient with Marfan's syndrome. Preoperative computed tomography and echocardiography showed acute aortic dissection (DeBakey type 1), severe aortic regurgitation, annuloaortic ectasia, and mild mitral regurgitation (MR). We performed artificial chordae implantation to the anterior mitral leaflet (AML) through the aortic root, followed by insertion of an aortic composite graft and replacement of the aortic arch. The patient is well 55 months after the operation, with minimal MR. We think that the transaortic approach is a good alternative for exposure and correction of the AML and its apparatus in special circumstances. PMID- 15338354 TI - Peptic ulcer-induced acute aortogastric fistula occurring 7 years after a pharyngogastrostomy following a resection for carcinoma of the esophagus: report of a case. AB - A 46-year-old woman underwent a pharyngogastrostomy, following a laryngoesophagectomy for esophageal carcinoma. Although she had been disease-free for 7 years, she subsequently was admitted to undergo a workup due to fever along with chest and back pain. A few days after admission, the patient suddenly vomited a large volume of blood and went into shock. Bleeding was stopped with a Sengstaken-Blakemore tube, and an emergency thoracotomy was performed. A fistula between the thoracic aorta and an ulcer of the gastric tube was identified. We decided to close the aortic lesion directly because the adhesions were extremely dense and her blood circulation was poor. One week later, we resected the thoracic part of the gastric tube, debrided the fistula, and wrapped the aortic lesion with a patch. However, on the 18th postoperative day, she developed massive hematemesis due to rupture of an infected pseudoaneurysm in the thoracic aorta and died. PMID- 15338355 TI - Primary malignant mesothelioma of the greater omentum: report of a case. AB - We report a rare case of primary malignant mesothelioma of the greater omentum. To our knowledge, only one other such case has been described in the English literature. The patient was a 61-year-old Taiwanese woman without any history of exposure to asbestos, who presented with lower back pain. Abdominal sonography and computed tomography showed a 12 x 9 x 9-cm(3) mass occupying the lower abdomen. Laparotomy revealed a tumor in the greater omentum, invading the posterior wall of the uterus, without diffuse mesenteric thickening or multiple small nodules in the peritoneum. We performed en bloc resection of the mass, which involved omentectomy, hysterectomy, and bilateral salpingo-oophorectomy. Microscopically, the tumor cells were arranged in a tubulopapillary pattern lined by a single layer of uniform, cuboidal cells. A pattern of sclerotic stroma with irregular glandular elements was also recognized. Immunohistochemically, the tumor cells showed strong positivity for calretinin. The final pathologic diagnosis was malignant mesothelioma. The patient did not receive chemotherapy or radiotherapy, and has remained in good health without any evidence of recurrence for almost 3 years since her operation. PMID- 15338356 TI - Long-term survival achieved by repeated resections of metachronous pulmonary and adrenal metastases of alpha-fetoprotein-producing gastric cancer: report of a case. AB - We report a case of long-term survival achieved by repeated resections of metastases from alpha-fetoprotein (AFP)-producing gastric cancer. A 59-year-old man initially underwent total gastrectomy with lymph node dissection and resection of the spleen and left adrenal gland, for an advanced AFP-producing gastric cancer. A solitary pulmonary metastasis was resected 2 years later, and then a right adrenal gland metastasis, an inferior vena cava thrombus, and another pulmonary metastasis were resected 6 months, 1 year, and 8 months after each other, respectively. Thus, the patient has survived for 7 years and no further evidence of disease was found at his last follow-up examination. To the best of our knowledge, this is the first case of AFP-producing gastric cancer resulting in metachronous solitary pulmonary and adrenal gland metastases, but not liver metastasis. We report this case to show that for selected cases, surgical resection of these metastases is feasible and may extend survival. PMID- 15338357 TI - Churg-Strauss syndrome manifesting as perforation of the small intestine: report of a case. AB - We report a case of Churg-Strauss syndrome (CSS) causing perforation of the small intestine. A 51-year-old woman was admitted with an asthma attack and paralysis of both legs. Intravenous predonisolone (40 mg) was given to relieve her asthma. Laboratory data on admission showed leukocytosis with hypereosinophilia and a high level of serum IgE. Neurological examination also revealed mononeurutis multiplex. Based on these findings, we diagnosed CSS, and oral corticosteroids were continued. On the 20th day after admission, she suffered sudden abdominal pain. Abdominal X-ray showed free air in the abdomen, suggesting perforation of the gastrointestinal tract. Emergency laparotomy revealed generalized peritonitis caused by a perforated ulcer of the ileum. The resected specimens contained a perforation and multiple nonperforated ulcers with an irregular shape on the mucosal surface. Histopathological examinations revealed angiitis of the small vessels surrounded by eosinophilic infiltration and granuloma, consistent with CSS. Considering the high risk of perforation of the gastrointestinal tract, including the small intestine, during corticosteroid treatment in patients with CSS, any abdominal pain or discomfort must be investigated carefully. PMID- 15338358 TI - Intestinal obstruction in the terminal ileum caused by an anomalous congenital vascular band between the mesoappendix and the mesentery: report of a case. AB - We report a case of intestinal obstruction caused by a congenital abnormal vascular band in a 17-year-old boy. The patient was admitted with acute colicky abdominal pain, and an emergency laparotomy revealed that the ileum was strangulated by a fibrous band with vessels about 2 mm in diameter and 7 cm in length, extending from the antemesenterium of the terminal ileum to the mesoappendix. The affected intestine was resected with the band and the appendix. Histologically, the fibrous band was composed of loose connective tissue containing arteries, veins, and nerve fibers, suggesting that it was congenital and originated from a remnant of the ventral mesentery in the embryonic period. There have been few reports of intestinal obstruction being caused by a congenital vascular band, especially in patients beyond the pediatric age group. PMID- 15338359 TI - Rectal duplication as an unusual cause of chronic perianal fistula in an adult: report of a case. AB - Duplication of the rectum is a rare embryologic event, but it should be considered as a possibility when perianal fistulas and abscesses remain resistant to conventional standard surgical treatment modalities over the long term. We report the case of a 57-year-old woman who underwent many operations over 30 years for persistent perianal fistulas. After radiological assay by computed tomography, fistulography, and barium enema studies, we performed surgery to remove a cystic mass in the retrorectal region, which was subsequently found to be a rectal duplication. The patient had an uneventful postoperative course and has been asymptomatic for 3 years. PMID- 15338360 TI - Localization of a colovesical fistula using a retrograde guide-wire: report of a case. AB - We describe an unconventional method of localizing a colovesical fistula by using a guide-wire, successfully carried out in a 45-year-old man with recurrent dysuria, pneumaturia, and suprapubic tenderness. First, we performed a cystoscopy to establish the fistulous tract in the bladder and passed the guide-wire through it. Next, we performed a colonoscopy, and the guide-wire was identified and brought out through the anus. This created a wire loop through the fistula. The transparietal catheter enabled us to detect the exact fistulous tract at laparotomy, making it possible to resect the inflamed colon and identify and resect the fistulous opening on the vesical wall. This technique allowed for a safer resection and a shorter operation time. PMID- 15338361 TI - Portal vein resection for a portal vein thrombus caused by nonfunctioning islet cell carcinoma: report of a case. AB - We report a case of nonfunctioning islet cell carcinoma of the pancreas causing a tumor thrombus in the portal vein. The patient was a 60-year-old woman whose presenting symptoms were abdominal pain, vomiting, and weight loss. We performed a subtotal pancreatectomy and splenectomy combined with partial resection of the portal vein. Histopathological studies confirmed the diagnosis of nonfunctioning islet cell carcinoma of the pancreas with a tumor thrombus in the portal vein. The patient's postoperative course was uneventful and she is doing well 25 months after the operation. PMID- 15338362 TI - Pregnancy-related pelvic girdle pain (PPP), I: Terminology, clinical presentation, and prevalence. AB - Pregnancy-related lumbopelvic pain has puzzled medicine for a long time. The present systematic review focuses on terminology, clinical presentation, and prevalence. Numerous terms are used, as if they indicated one and the same entity. We propose "pregnancy-related pelvic girdle pain (PPP)", and "pregnancy related low back pain (PLBP)", present evidence that the two add up to "lumbopelvic pain", and show that they are distinct entities (although underlying mechanisms may be similar). Average pain intensity during pregnancy is 50 mm on a visual analogue scale; postpartum, pain is less. During pregnancy, serious pain occurs in about 25%, and severe disability in about 8% of patients. After pregnancy, problems are serious in about 7%. The mechanisms behind disabilities remain unclear, and constitute an important research priority. Changes in muscle activity, unusual perceptions of the leg when moving it, and altered motor coordination were observed but remain poorly understood. Published prevalence for PPP and/or PLBP varies widely. Quantitative analysis was used to explain the differences. Overall, about 45% of all pregnant women and 25% of all women postpartum suffer from PPP and/or PLBP. These values decrease by about 20% if one excludes mild complaints. Strenuous work, previous low back pain, and previous PPP and/or PLBP are risk factors, and the inclusion/exclusion of high-risk subgroups influences prevalence. Of all patients, about one-half have PPP, one third PLBP, and one-sixth both conditions combined. Overall, the literature reveals that PPP deserves serious attention from the clinical and research communities, at all times and in all countries. PMID- 15338363 TI - Measuring immunity in viral hepatitis. PMID- 15338364 TI - Association of Lewis and Secretor gene polymorphisms and Helicobacter pylori seropositivity among Japanese-Brazilians. AB - BACKGROUND: Secretor ( Se) and Lewis ( Le) genes are involved in the synthesis of Lewis b (Le(b)) and type I antigens throughout the body, especially in the epithelial cells of gastric mucosa. Helicobacter pylori can attach to the gastric epithelial cells with the blood group antigen-binding adhesin, which binds to Le(b) or H type I carbohydrate structures. In a previous study, a marked association between H. pylori seropositivity and polymorphism of the Se and Le genes was observed among Japanese outpatients of a gastroenterology clinic. The present work aims to investigate the associations between Se and Le gene polymorphisms and H. pylori infection among Japanese-Brazilians. METHODS: The subjects consisted of 942 healthy volunteer Japanese-Brazilians, who were tested for the presence of anti- H. pylori IgG antibodies and genotyped for Se and Le polymorphisms. RESULTS: The sex-age-adjusted odds ratios (aORs) for H. pylori seropositivity were 0.99 for the Sese genotype relative to the SeSe genotype (95% confidence interval [CI], 0.73-1.33), and 1.03 for sese relative to SeSe (95% CI, 0.71-1.48). On the other hand, the aOR for the subjects with the le allele ( Lele or lele) relative to the LeLe genotype was 1.48 (95% CI, 1.07-1.79). When the Se and Le genotypes were analyzed in combination according to risk group, no statistically significant association was observed. CONCLUSIONS: These results are inconsistent with previous work and may have been modulated by an external factor or some other unidentified factor. Japanese-Brazilians are genotypically the same as Japanese, but their lifestyle is adapted to that of Brazil. Further investigations are necessary to clarify this influence on susceptibility to H. pylori infection. PMID- 15338365 TI - Expression of a 72-kDa heat shock protein, and its cytoprotective function, in gastric mucosa in cirrhotic rats. AB - BACKGROUND: Portal hypertensive gastropathy (PHG) is a clinical entity that is observed frequently in patients with liver cirrhosis. In PHG, gastric mucosa is highly susceptible to mucosal injury caused by noxious agents. Many studies, including ours, have reported that a 72-kDa heat shock protein (HSP72) has a crucial cytoprotective function in gastric mucosa. In this study, we investigated the expression and cytoprotective effect of HSP72 on gastric mucosa in portal hypertensive rats. METHODS: PHG was produced by bile duct ligation (BDL) or carbon tetrachloride administration in male Sprague-Dawley rats. The expression of HSP72 in the gastric mucosa was evaluated by Western blotting. Induction of gastric mucosal HSP72 by 6-h water-immersion stress was compared between cirrhotic and control rats. Also, mucosal protective abilities against hydrochloric acid (HCl; 0.6 N) following pretreatment with water-immersion stress to induce HSP72 were studied in both groups. RESULTS: Portal venous pressure was significantly higher in cirrhotic rats compared with control rats ( P < 0.05). Baseline expression (before water-immersion stress) of mucosal HSP72 was significantly lower in cirrhotic rats compared with control rats. HCl-induced gastric mucosal lesions were significantly suppressed in control rats compared with cirrhotic rats, especially when HSP72 was preinduced by water-immersion stress. CONCLUSIONS: These findings suggest that HSP72 in the gastric mucosa plays a crucial role with respect to cytoprotection; the induction of HSP72 may provide therapeutic strategies for protection against mucosal injury in PHG. PMID- 15338366 TI - The prevalence of Helicobacter pylori in Japanese children with gastritis or peptic ulcer disease. AB - BACKGROUND: Although Helicobacter pylori infection is typically acquired in childhood, the role of H. pylori infection in gastroduodenal diseases in childhood remains to be defined. The purpose of this study was to evaluate the prevalence of H. pylori infection in children with gastritis, duodenal ulcer, and gastric ulcer. METHODS: This was a retrospective analysis of 283 Japanese children (mean age, 11.5 years) with non-nodular gastritis ( n = 73), nodular gastritis ( n = 67), duodenal ulcer ( n = 100), and gastric ulcer ( n = 43). H. pylori status was based on biopsy tests. Clinical symptoms at the time of endoscopy were analyzed with regard to a possible association with the infection. RESULTS: The prevalence of H. pylori in non-nodular gastritis, nodular gastritis, duodenal ulcer, and gastric ulcer was 28.8%, 98.5%, 83.0%, and 44.2%, respectively. H. pylori was significantly linked to duodenal ulcer and gastric ulcers in the age group of 10-16 years, but not in the age group of 9 years and under. In children with H. pylori infection, nodular gastritis was observed in 26.3% of gastric ulcer patients and in 74.7% of duodenal ulcer patients ( P < 0.001). H. pylori infection was significantly associated with the prevalence of anemia ( P < 0.05). CONCLUSIONS: H. pylori is the most important causal factor for the development of duodenal ulcer in childhood. While H. pylori infection appears to be a risk factor in gastric ulcer, other causes are responsible for most cases. Nodular gastritis is the most common type of H. pylori gastritis in childhood. Chronic infection with H. pylori is associated with anemia. PMID- 15338367 TI - Characteristics of gastric B-cell lymphoma of mucosa-associated lymphoid tissue type involving multiple organs. AB - BACKGROUND: Mucosa-associated lymphoid tissue (MALT) lymphoma involving multiple organs is not well characterized. METHODS: To obtain a better understanding of gastric MALT lymphoma with multiple organ involvement, we compared Helicobacter pylori infection status; response to eradication therapy; prevalence of genetic abnormality, including t(11;18)(q21;q21); and clonality of tumor cells between patients with gastric MALT lymphoma with and without multiple organ involvement. RESULTS: Of 54 patients with only gastric involvement, 51 were positive for H. pylori (94.4%), whereas only 3 of 9 patients with multiple organ involvement had the infection (33.3%). Among those who received eradication therapy, the remission rate in patients with only gastric involvement was significantly higher than that in patients with multiple organ involvement (71.4% vs 33.3%; P < 0.05). The positive rate of API2-MALT1 fusion transcripts was significantly greater in patients with multiple lesions with either single or multiple organ involvement than in those with a single gastric lesion (71.4% vs 12.5%; P < 0.05), but 2 of 2 patients with multiple gastric lesions had the fusion transcripts. Of 5 patients with multiple organ involvement tested for clonality of the CDR3 region, distinct clones were detected in lymphoma lesions in different organs in 3 patients, whereas the identical clone was present in the different lesions in the remaining 2 patients. CONCLUSIONS: Gastric MALT lymphoma with multiple organ involvement is often associated with API2-MALT1 fusion transcripts. Moreover, H. pylori infection is rare in patients with multiple organ involvement as compared to those with only gastric involvement. Patients with gastric MALT lymphoma with multiple organ involvement who are positive for H. pylori are resistant to eradication therapy. PMID- 15338368 TI - Endoscopic ultrasound-guided fine-needle aspiration biopsy for the diagnosis of gastrointestinal stromal tumors in the stomach. AB - BACKGROUND: For the diagnosis of gastric submucosal tumors (SMTs), endoscopic ultrasound (EUS) alone does not reveal the complete pathology, such as the degree of malignancy, and EUS-guided fine-needle aspiration biopsy (EUS-FNAB) has been reported to be more useful. Recently, most cases initially diagnosed as leiomyosarcomas have received further study with immunohistochemical staining and have been given the new diagnosis of gastrointestinal stromal tumors (GISTs). The degree of malignancy of GISTs differs widely in clinical aspects. In this study, we examined whether EUS-FNAB was useful in diagnosing GISTs and differentiating their degrees of malignancy. METHODS: From January 1997 to March 2002, 21 cases of gastric GISTs were diagnosed from the immunohistochemical staining of specimens resected at Aichi Cancer Center Hospital. Of these 21 patients, 14 (5 with high-grade malignancy and 9 with low-grade malignancy) underwent EUS-FNAB preoperatively, and were examined further: their EUS-FNAB specimens were submitted for additional immunohistochemical testing. RESULTS: The EUS-FNAB specimens from all patients were positive for c-kit and CD34 immunohistochemical testing, coinciding with the staining results of the resected specimens. The MIB 1 labeling indices in specimens of high-grade malignancy were significantly higher than those of low-grade malignancy. If we assumed that a tumor with an MIB 1 labeling index of more than 5% was a high-grade malignancy, the diagnostic accuracy was 85.7%. CONCLUSIONS: The EUS-FNAB procedure is a useful tool for diagnosing GISTs of the stomach with immunohistochemical staining. When used with MIB-1 staining, the procedure may indicate GIST prognosis and influence decisions regarding therapeutic strategies. PMID- 15338369 TI - Infection and dysfunction of circulating blood dendritic cells and their subsets in chronic hepatitis C virus infection. AB - BACKGROUND: To understand interactions between dendritic cells (DCs) and viruses, in vitro-cultured monocyte-derived DCs are usually used for functional analyses. However, several recent studies indicate that circulating blood DCs are different from monocyte-derived DCs, both phenotypically and functionally. Indeed, circulating DCs act as functional antigen-presenting cells in vivo. This study was conducted to evaluate the function of circulating blood DCs in patients with chronic hepatitis C and to examine whether circulating DCs from these patients were infected by hepatitis C virus (HCV). METHODS: The phenotypes and biological functions of circulating DCs from patients with chronic hepatitis C ( n = 27), patients with non-HCV chronic liver disease ( n = 7), and normal volunteers ( n = 13) were analyzed. The presence of the HCV genome sequence in circulating blood DCs and in subsets of circulating DCs (myeloid DCs and plasmacytoid DCs) in patients with chronic hepatitis C was assessed. RESULTS: The stimulatory capacity of circulating DCs was significantly reduced in patients with chronic hepatitis C compared to patients with non-HCV chronic liver diseases and normal controls ( P < 0.01). HCV RNA was identified in the overall population of circulating DCs, and in myeloid DCs and plasmacytoid DCs. Nucleotide sequences of the 5' non-coding region of HCV RNA showed marked differences between paired samples of circulating DCs and sera from the same patients. CONCLUSIONS: Our results indicate the dysfunction and infection of circulatory blood DCs in chronic HCV infection. This may compromise the capacity of patients with hepatitis C to induce an effective antiviral immune response. PMID- 15338370 TI - Expression of gap junction protein connexin32 in chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. AB - BACKGROUND: The gap junction (GJ) plays important roles in the maintenance of tissue homeostasis, the control of cell growth and differentiation, and the prevention of experimental hepatocarcinogenesis. In this study, we examined the relationship between the expression of the GJ protein connexin (Cx) 32 in 24 human hepatocellular carcinomas (HCCs) and 29 non-carcinomatous liver specimens (NCLs) of 31 patients. METHODS: An immunohistochemical study of Cx32 was done in 24 HCCs and 29 NCLs from 31 patients who had undergone hepatic resection. RESULTS: The Cx32 expression decreased gradually as the disease progressed to cirrhosis and HCC. In all Cx32 positive HCCs, the expression was mostly recognized in cytoplasm, not only on the cell membrane. This internalization of Cx32 was also recognized in liver specimens showing hepatitis and cirrhosis, although it was less frequent than in the HCCs. CONCLUSIONS: These findings suggest the possibility that changes in both the amount and the distribution of Cx32 may be implicated in human hepatocarcinogenesis. PMID- 15338371 TI - Prognostic indicators of breakthrough hepatitis during lamivudine monotherapy for chronic hepatitis B virus infection. AB - BACKGROUND: Breakthrough hepatitis (BTH), defined as a flare of transaminases alanine aminotransferase [ALT]) can occur during lamivudine monotherapy for hepatitis B virus (HBV) infection. There have been many reports of lamivudine resistant mutations within the C domain of the viral reverse transcriptase; however, the appearance of these mutants is not necessarily correlated with BTH during lamivudine therapy. METHODS AND RESULTS: Entire serial HBV genomic sequences before and during lamivudine therapy for 4 patients with BTH and 1 patient without BTH were analyzed and showed changes in the pre-S region. These changes may be associated with ALT flares. Further investigation in a cohort of 36 patients with a median treatment period of 25 months showed that 21 patients had a rise in HBV-DNA titer, of whom 18 had BTH. Univariate statistical analyses showed that possible prognostic indicators for the occurrence of BTH were pre-S deletions ( P = 0.03) and L180M/M204L mutations ( P = 0.04). By multivariate Cox regression analyses, significant variables were pre-S deletions (hazard ratio, 0.17; 95% confidence internal (CI), 0.044-0.66) and precore mutations (hazard ratio, 5.70; 95% CI, 1.74-18.71) prior to the commencement of lamivudine monotherapy. Interestingly, BTH occurred after the selection of the wild-type species in the pre-S region during lamivudine monotherapy. CONCLUSIONS: These results suggest that patients with HBV pre-S deletion mutants should be monitored carefully during lamivudine therapy. PMID- 15338372 TI - Interferon monotherapy for patients with chronic hepatitis C and normal serum aminotransferase levels at commencement of treatment. AB - BACKGROUND: Approximately 30% of patients with chronic hepatitis C have normal serum alanine amino transferase (ALT) levels. While interferon (IFN) monotherapy is approved for patients with chronic hepatitis C infection, the effectiveness of such therapy for chronic hepatitis C patients with normal ALT levels at commencement of treatment remains poorly understood. METHODS: Ninety-four individuals (M/F, 54 : 40; median age, 46 years) with normal ALT levels (< 50 IU/l) at the commencement of treatment who were positive for both anti-hepatitis C virus (HCV) and serum HCV-RNA were studied. Among this group, 18 individuals (M/F, 9 : 9; median age, 50 years) had had persistently normal ALT levels for at least 3 months prior to treatment. All patients received their first course of IFN therapy in this study. RESULTS: Forty-three (45.7%) of 94 individuals had lost serum HCV-RNA at 6 months after cessation of therapy (complete response; CR). The proportion of patients with genotype 2a and HCV-RNA level over 1 Meq/ml who showed CR was significantly lower in those with normal ALT levels than in those with elevated ALT levels (23.8% vs 55.6%; P = 0.0189). Two patients who had persistently normal ALT levels and HCV-RNA level over 1 Meq/ml were nonresponders (NR) and had ALT flare-ups after IFN therapy. Patients with HCV-RNA levels of less than 1 Meq/ml did not show differential responses based on ALT levels. CONCLUSIONS: Our data suggest that IFN therapy is effective for patients with normal ALT levels and less than 1 Meq/ml HCV-RNA. Thus, such patients should be considered for curative IFN therapy. PMID- 15338373 TI - Genetic analysis of the glutathione s-transferase genes MGST1, GSTM3, GSTT1, and GSTM1 in patients with hereditary pancreatitis. AB - BACKGROUND: Specific mutations in the cationic trypsinogen gene ( PRSS1) are disease-causing in patients with hereditary pancreatitis, but the genetic background still remains mysterious in about 40% of patients with the disease. It has been suggested that oxidative stress contributes to pancreatic damage. The glutathione s-transferases (GSTs) represent major detoxification enzymes that protect cells from oxidative stress. METHODS: In the present study we tested whether mutations in the MGST1 and GSTM3 genes or common deletions in the GSTT1 and GSTM1 genes are associated with hereditary pancreatitis. We analyzed the entire coding region of MGST1 and GSTM3 in 30 patients that were tested negative for PRSS1 mutations, and we studied 55 controls. For GSTT1 and GSTM1, we investigated 75 hereditary pancreatitis patients who had been tested negative for PRSS1 mutations, 135 hereditary pancreatitis patients with a PRSS1 mutation, and 183 controls. Patients were further subclassified with regard to age of onset of disease as a marker of severity. RESULTS: No mutation was found in the MGST1 gene. In the GSTM3 gene, we detected a homozygous 670G > A polymorphism (V224I) with similar frequencies in patients and controls. We found no difference in the frequencies of the GSTT1 and GSTM1 null genotypes between patients and controls, and we detected no differences in age of onset in patients with or without GSTT1 and GSTM1 deletions. CONCLUSIONS: We conclude that genetic alterations in the MGST1, GSTM3, GSTT1, and GSTM1 genes do not play a dominant role in hereditary pancreatitis. PMID- 15338374 TI - Primary, solitary, adult T-cell leukemia / lymphoma of the descending colon. AB - We herein report a patient with adult T-cell leukemia/lymphoma (ATLL) of the descending colon. A 64-year-old man was admitted to our hospital complaining of left lower abdominal pain. Endoscopic examination revealed an ulcerative tumor in the descending colon that was diagnosed as T-cell lymphoma by biopsy. Neither distant organ metastasis nor lymph node swelling was observed by radiographic examinations. Curative excision with left hemicolectomy and regional lymph node dissection was performed. Surgical sections contained ulcerative and superficially elevated lesions; these were continuous with each other. Histological examination revealed diffuse proliferation of medium-sized abnormal lymphoid cells. Immunohistochemically, these lymphoid cells were positive for UCHL-1/CD45RO and CD3 and negative for CD79a, indicating that the tumor was a primary malignant T-cell lymphoma of the descending colon. Integration of the proviral DNA of human T-lymphotropic virus type 1 (HTLV-1) was confirmed by Southern blotting analysis. PMID- 15338375 TI - Interstitial pneumonia induced by combined intraarterial 5-fluorouracil and subcutaneous interferon-alpha therapy for advanced hepatocellular carcinoma. AB - Previously we reported combined chemo-immunotherapy, using interferon (IFN)-alpha and 5-fluorouracil (5-FU) for patients with advanced hepatocellular carcinoma (HCC), and this regimen improved the prognosis. Recently, we experienced an HCC patient who died of severe interstitial pneumonia during the combined IFN-alpha and 5-FU therapy. This is the first report of the occurrence of interstitial pneumonia during combined IFN-alpha and 5-FU treatment. A 60-year-old-man was admitted to Osaka University Hospital to receive systemic chemo-immunotherapy for recurrent HCC. In the second week of the chemo-immunotherapy, he showed a decreased level of consciousness, and respiratory insufficiency. Emergency roentgenogram revealed diffuse infiltration in both lungs. Respiratory dysfunction due to interstitial pneumonia was suspected, and steroid pulse therapy was started. However, the patient showed respiratory failure, and he died 32 days after the start of the therapy. Autopsy findings showed atelectasis in the bilateral lungs, which showed elastic hard solidity and a dark red color; esophageal varices were also shown, and there was cirrhosis with a large tumor in the liver. Microscopically, the alveolar wall showed marked fibrous thickness and moderate inflammatory change, which is consistent with acute interstitial pneumonia, and the acute pulmonary change was suspected to have been the cause of death. The association of IFN with the development of interstitial pneumonia has been reported. However, the prognosis of IFN-induced interstitial pneumonia has mostly been favorable when the medication was discontinued. It has been postulated that interstitial pneumonia induced by the combination of IFN and 5-FU may be therapy-resistant. The combination of IFN-alpha and 5-FU is a useful therapy for patients with advanced HCC, such as that with portal vein invasion or multiple metastatic foci. Thus, interstitial pneumonia in these patients should be carefully managed. PMID- 15338376 TI - Pancreatic metastasis of dermatofibrosarcoma protuberans. AB - Dermatofibrosarcoma protuberans (DFSP) is a relatively rare skin tumor that is considered to have intermediate malignancy; it demonstrates frequent local recurrence, but systemic metastasis is rare. We report a 49-year-old woman with pancreatic metastasis of DFSP who underwent total pancreatectomy with partial resection of the portal vein. Except for our patient, only two other cases of pancreatic metastasis of DFSP have been reported in the literature, to our knowledge. Radical resection may be considered for pancreatic metastasis of DFSP when there are no other metastatic lesions. PMID- 15338377 TI - Negative immunohistochemical staining of p53 protein does not always reflect wild type p53 gene in cancer cells. PMID- 15338378 TI - Video capsule endoscopy for evaluating obscure gastrointestinal bleeding and suspected small-bowel pathology. PMID- 15338379 TI - Fulminant hepatic failure caused by malignant melanoma of unknown primary origin. PMID- 15338380 TI - Why are the results inconsistent on relationships between Se and Le genotypes and continuous infection of H. pylori? PMID- 15338381 TI - Helicobacter pylori infection in childhood. PMID- 15338382 TI - Distinct susceptibility of dendritic cell subsets to hepatitis C virus infection: a plausible mechanism of dendritic cell dysfunction. PMID- 15338383 TI - Events occurring at the time of breakthrough hepatitis during lamivudine treatment for chronic hepatitis B. PMID- 15338384 TI - Use of complementary and alternative medicine by patients with breast cancer: observations from a health-care survey. AB - GOALS OF THE WORK: The aim of this study was to assess the period prevalence and identify predictors of the use of complementary and alternative medicine (CAM) among women with breast cancer. PATIENTS AND METHODS: In a cross-sectional study, 263 women participating in a quality of life survey reported on CAM use. Differences between CAM users and nonusers were evaluated using logistic regression. RESULTS: About 3 years after diagnosis the period prevalence of CAM use among women with breast cancer was 36%. Younger women reported usage of CAM more often than older women. Associations between remote disease, pain and psychosocial variables were found. Involvement in self-help groups and active participation in leisure activities were associated with CAM use; this may indicate differences in the patients' ability to cope with the disease. Most patients were informed of CAM by a physician. CONCLUSIONS: Involvement in self help groups and leisure activities were associated with CAM use, indicating differences in the patients' ability to cope. Further research is necessary to evaluate the safety of CAM and to determine the value of CAM in health care. PMID- 15338385 TI - Effect of aerobic exercise and relaxation training on fatigue and physical performance of cancer patients after surgery. A randomised controlled trial. AB - Fatigue is a frequent problem after surgical treatment of solid tumours. Aerobic exercise and psychosocial interventions have been shown to reduce the severity of this symptom in cancer patients. Therefore, we compared the effect of the two therapies on fatigue in a randomised controlled study. Seventy-two patients who underwent surgery for lung (n=27) or gastrointestinal tumours (n=42) were assigned to an aerobic exercise group (stationary biking 30 min five times weekly) or a progressive relaxation training group (45 min three times per week). Both interventions were carried out for 3 weeks. At the beginning and the end of the study, we evaluated physical, cognitive and emotional status and somatic complaints with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core Module (EORTC-QLQ-30) questionnaire, and maximal physical performance with an ergometric stress test. Physical performance of the training group improved significantly during the programme (9.4+/-20 watts, p=0.01) but remained unchanged in the relaxation group (1.5+/-14.8 watts, p=0.37). Fatigue and global health scores improved in both groups during the intervention (fatigue: training group 21%, relaxation group 19%; global health of both groups 19%, p for all < or =0.01); however, there was no significant difference between changes in the scores of both groups (p=0.67). We conclude that a structured aerobic training programme improves the physical performance of patients recovering from surgery for solid tumours. However, exercise is not better than progressive relaxation training for the treatment of fatigue in this setting. PMID- 15338386 TI - Prevalence of weather sensitivity in Germany and Canada. AB - Several studies have shown that atmospheric conditions can affect well-being or disease, and that some individuals seem to be more sensitive to weather than others. Since epidemiological data on the prevalence of weather-related health effects are lacking, two representative weather sensitivity (WS) surveys were conducted independently in Germany and Canada. The objectives of this paper are: (1) to identify the prevalence of WS in Germany and Canada, (2) to describe weather-related symptoms and the corresponding weather conditions, and (3) to compare the findings in the two countries. In Germany 1,064 citizens (age >16 years) were interviewed in January 2001, and in Canada 1,506 persons (age >18 years) were interviewed in January 1994. The results showed that 19.2% of the German population thought that weather affected their health "to a strong degree," 35.3% that weather had "some influence on their health" (sum of both = 54.5% weather sensitive), whereas the remaining 45.5% did not consider that weather had an effect on their health status. In Canada 61% of the respondents considered themselves to be sensitive to the weather. The highest prevalence of WS (high + some influence) in Germans was found in the age group older than 60 years (68%), which was almost identical in the Canadian population (69%). The highest frequencies of weather-related symptoms were reported in Germany for stormy weather (30%) and when it became colder (29%). In Canada mainly cold weather (46%), dampness (21%) and rain (20%) were considered to affect health more than other weather types. The most frequent symptoms reported in Germany were headache/migraine (61%), lethargy (47%), sleep disturbances (46%), fatigue (42%), joint pain (40%), irritation (31%), depression (27%), vertigo (26%), concentration problems (26%) and scar pain (23%). Canadian weather-sensitive persons reported colds (29%), psychological effects (28%) and painful joints, muscles or arthritis (10%). In Germany 32% of the weather-sensitive subjects reported themselves to be unable to do their regular work because of weather related symptoms at least once in the previous year, and 22% of them several times. Co-morbidity was significantly higher in weather-sensitive subjects both in Germany and Canada. These results clearly showed the important impact of WS on public health and the economy. These findings prompted us to start studies on the causal factors of weather-related health effects. PMID- 15338387 TI - Note of caution for the use of sodium nitroprusside in neonatal hypertension. PMID- 15338388 TI - Tumor necrosis factor-alpha blocking agent as a treatment for nephrotic syndrome. AB - We report a 13-year-old boy with refractory nephrotic syndrome (minimal change with mesangial proliferation) who failed the standard treatment protocols. There was some temporary response to large steroid doses, but even the Mendoza protocol could not induce remission. We show suppression of the proteinuria with Infliximab (Remicade) with tapering of steroids. Serial serum levels of tumor necrosis factor (TNF)-alpha are shown and discussed. We suggest studying the TNF alpha blocking agents as optional treatment for nephrotic syndrome. PMID- 15338390 TI - Bone disease in nephrotic syndrome--prevention is better than cure. PMID- 15338391 TI - Postnatal renal development of rats from mothers that received increased sodium intake. AB - The newborn rat kidney is not fully developed until approximately 12 days after birth. Several lines of evidence suggest that angiotensin II (AII) participates in the postnatal development of the kidney. The aim of the present study was to analyze proliferating cell nuclear antigen (PCNA), fibronectin, alpha-smooth muscle-actin (alpha-SM-actin), and AII expression in renal cortex during development in rats born to mothers that received a normal (control) or increased (experimental) sodium intake during pregnancy. Ninety Wistar rats aged 1, 7, 15, and 30 days from the control and experimental groups were killed and the kidneys removed for histological and immunohistochemical studies. The results showed higher fibronectin, alpha-SM-actin, PCNA, and AII expression in the glomerular and tubulointerstitial areas of the renal cortex of 1- and 7-day-old animals, which decreased with renal development. The animals from the experimental group showed at 1 day of age a decrease in alpha-SM-actin, fibronectin, PCNA, and AII expression compared with controls of the same age ( P<0.05). In conclusion, our data show that increased sodium intake during pregnancy induces a reduction of alpha-SM-actin, fibronectin, and PCNA expression in the renal cortex tubulointerstitium and glomeruli of neonatal rats. These alterations may be related to the decrease of AII expression also observed in the renal cortex from these animals. PMID- 15338392 TI - Renovascular hypertension due to antithrombin deficiency in childhood. AB - Inherited antithrombin deficiency generally causes a predisposition toward vascular thrombus above the age of 15 years. A 1-year-old boy developed renal hypertension caused by renal artery obstruction due to thrombus formation. This thrombus formation was attributed to antithrombin deficiency caused by a novel SERPINCI gene mutation (AT III Akita, M352R). This suggests that antithrombin deficiency can cause renal artery obstruction, inducing renal hypertension through vascular thrombosis even in children. PMID- 15338393 TI - Kimura disease with advanced renal damage with anti-tubular basement membrane antibody. AB - Kimura disease (KD) is an autoimmune eosinophilic granulomatous disorder with generalized lymphadenopathy. A handful of pediatric patients with renal disease have been described, none of whom have been African-American (AA). We present an AA boy with KD and nephrotic syndrome (NS). Two months after stopping steroids, fever, asthma, eczema, and proteinuria recurred. His NS did not relapse but his platelet count decreased to 51,000/microl (x10(6)/l). On restarting prednisone, his platelet count normalized. A kidney biopsy revealed 23 of 37 glomeruli obsolescent and advanced damage with over 50% of cortical tissue replaced by interstitial fibrosis and chronic inflammation. Glomerular immunofluorescence was largely negative; very intense linear anti-tubular basement membrane (TBM) deposits of IgA, IgG, C3, and C4 were noted. At present, 36 months from onset, serum creatinine is 1.2 mg/dl (106 micromol/l). We present a 4-year-old AA boy with KD, NS, relapsing thrombocytopenia, and renal damage with anti-TBM antibody. PMID- 15338394 TI - Creation of a novel hemodialysis bridge graft to extend the life of an adolescent. AB - All conventional dialysis access options were exhausted in a 19-year-old girl with end-stage renal disease (ESRD). Her course was medically and psychosocially complicated. To save her life, a subclavian artery to right atrium hemodialysis bridge graft was created so that she could be dialyzed successfully. Adolescents and young adults confronted with the exhaustion of dialysis access and transplant options face extremely difficult management decisions. Attention to the multi disciplinary needs of these patients and a combined effort by medical, surgical, psychological, and social work teams are necessary to address properly these dilemmas. PMID- 15338396 TI - Megalin and proximal renal tubular dysfunction in Dent disease. PMID- 15338397 TI - Cardiac arrhythmias due to severe hypokalemia in a patient with classic Bartter disease. AB - We report a young girl with classic Bartter disease (type III) with severe hypokalemia (< or = 2.0 mmol/l) who developed a prolonged heart rate-corrected QT interval of 510 ms (upper reference 430 ms) and ST segment depression in all leads. Holter electrocardiography was performed (with a plasma potassium level of 2.0 mmol/l) and it disclosed a stable sinus rhythm, a prolonged correct QT interval, more-evident ST segment depression during an increase in heart rate, a few single premature ventricular complexes, and nocturnal conduction abnormalities such as second-degree atrioventricular block 2:1. In the light of these results, the treatment was modified by increasing indomethacin from 1.5 to 3 mg/kg per day and adding spironolactone at a dose of 5 mg/kg per day. After 10 days, plasma potassium levels increased to 2.7 mmol/l and electrocardiographic abnormalities regressed. No other cardiac abnormalities were noted when the serum potassium was maintained > 2.5 mmol/l. In conclusion, this case report supports the link between arrhythmic events and chronic renal hypokalemic alkalosis in renal tubular disorders. We highlight the importance of standardizing the use of rest electrocardiography and 24-h Holter monitoring to diagnose arrhythmic events in children with severe hypokalemic renal disorders, especially in those with a plasma potassium < 2.5 mmol/l. The importance of beginning early medical treatment, to improve plasma potassium levels and reverse cardiac abnormalities, is emphasized. PMID- 15338398 TI - No evidence for genotype/phenotype correlation in NPHS1 and NPHS2 mutations. AB - Primary steroid-resistant nephrotic syndrome (SRNS) is characterized by childhood onset of proteinuria and progression to end-stage renal disease. In 26% of cases it is caused by recessive mutations in NPHS2 (podocin). Congenital nephrotic syndrome (CNS) is caused by mutations in NPHS1 (nephrin) or NPHS2. In three families mutations in NPHS1 and NPHS2 had been reported to occur together, and these tri-allelic mutations were implicated in genotype/phenotype correlations. To further test the hypothesis of tri-allelism, we examined a group of 62 unrelated patients for NPHS1 mutations, who were previously shown to have NPHS2 mutations; 15 of 62 patients had CNS. In addition, 12 CNS patients without NPHS2 mutation were examined for NPHS1 mutations. Mutational analysis yielded three different groups. (1) In 48 patients with two recessive NPHS2 mutations (11 with CNS), no NPHS1 mutation was detected, except for 1 patient, who had one NPHS1 mutation only. This patient was indistinguishable clinically and did not have CNS. (2) In 14 patients with one NPHS2 mutation only (4 with CNS), we detected two additional recessive NPHS1 mutations in the 4 patients with CNS. They all carried the R229Q variant of NPHS2. The CNS phenotype may be sufficiently explained by the presence of two NPHS1 mutations. (3) In 12 patients without NPHS2 mutation (all with CNS), we detected two recessive NPHS1 mutations in 11 patients, explaining their CNS phenotype. We report ten novel mutations in the nephrin gene. Our data do not suggest any genotype/phenotype correlation in the 5 patients with mutations in both the NPHS1 and the NPHS2 genes. PMID- 15338413 TI - Asymmetric larval interactions between introduced and indigenous ladybirds in North America. AB - Understanding the mechanisms that result in the success of introduced species will contribute to predicting future invasions and managing invaded systems. We examined interactions between larvae of two predatory ladybird species recently introduced to North America, Coccinella septempunctata (CS) and Harmonia axyridis (HA), and two indigenous ladybirds, Coccinella transversoguttata (CT) and Hippodamia convergens (HC). By pairing young and old larvae in the laboratory at low and high levels of aphid availability, we assessed the degree of asymmetry in intraguild predation (IGP), the strength of competitive effects on growth and development of larvae escaping predation, and the nature of attack and escape behavior among the species. Interactions were generally asymmetric, with larvae of introduced species acting most frequently as intraguild predators and larvae of indigenous species serving most frequently as intraguild prey (the two Coccinella spp., however, preyed on each other at similar rates). Because they were especially aggressive and because other larvae were least successful in escaping their attacks, larvae of HA had stronger negative effects on larvae of the two indigenous species than did larvae of CS. Such negative effects, expressed most strongly when aphid availability was low, were especially adverse for the smaller of the two indigenous species, HC. In general, older larvae interacted with each other more strongly than young larvae did, and older larvae had especially strong negative effects on young larvae when interactions occurred between age classes. Our results suggest that HA more than CS may represent a threat to indigenous ladybirds as an intraguild predator, and that IGP in turn may play a stronger role for HA than for C. septempunctata in promoting the successful invasion of North America. PMID- 15338414 TI - Precipitation pulses and carbon fluxes in semiarid and arid ecosystems. AB - In the arid and semiarid regions of North America, discrete precipitation pulses are important triggers for biological activity. The timing and magnitude of these pulses may differentially affect the activity of plants and microbes, combining to influence the C balance of desert ecosystems. Here, we evaluate how a "pulse" of water influences physiological activity in plants, soils and ecosystems, and how characteristics, such as precipitation pulse size and frequency are important controllers of biological and physical processes in arid land ecosystems. We show that pulse size regulates C balance by determining the temporal duration of activity for different components of the biota. Microbial respiration responds to very small events, but the relationship between pulse size and duration of activity likely saturates at moderate event sizes. Photosynthetic activity of vascular plants generally increases following relatively larger pulses or a series of small pulses. In this case, the duration of physiological activity is an increasing function of pulse size up to events that are infrequent in these hydroclimatological regions. This differential responsiveness of photosynthesis and respiration results in arid ecosystems acting as immediate C sources to the atmosphere following rainfall, with subsequent periods of C accumulation should pulse size be sufficient to initiate vascular plant activity. Using the average pulse size distributions in the North American deserts, a simple modeling exercise shows that net ecosystem exchange of CO2 is sensitive to changes in the event size distribution representative of wet and dry years. An important regulator of the pulse response is initial soil and canopy conditions and the physical structuring of bare soil and beneath canopy patches on the landscape. Initial condition influences responses to pulses of varying magnitude, while bare soil/beneath canopy patches interact to introduce nonlinearity in the relationship between pulse size and soil water response. Building on this conceptual framework and developing a greater understanding of the complexities of these eco-hydrologic systems may enhance our ability to describe the ecology of desert ecosystems and their sensitivity to global change. PMID- 15338415 TI - The importance of seed reserves for seedling performance: an integrated approach using morphological, physiological, and stable isotope techniques. AB - To investigate how seed reserves affect early seedling performance, we conducted a factorial greenhouse experiment using Lithocarpus densiflora (Tanoak). Seedlings were grown from large (5.8+/-0.7 g) and small (3.2+/-0.4 g) seeds and, following shoot emergence, seeds were either removed or left attached. Seedlings were harvested for quantification of biomass and delta13C at seven time periods following seed removal (2, 4, 8, 16, 32, 64, 128 days) and seedling photosynthesis was measured three separate time periods (2-4, 49-82, 95-128 days after seed removal). Biomass increased for all seedlings, but the increase was significantly larger for seedlings with attached seeds than with removed seeds. Seed removal just after shoot emergence significantly decreased seedling biomass, but seed removal 64 days after shoot emergence had no effect on seedling biomass. Seedling photosynthesis per unit leaf area varied by time and seed presence, but not by seed size. At the first period, seedlings with attached seeds had significantly higher photosynthetic rates than seedlings with removed seeds, at the second period there was no effect of seed removal, and at the third time period seedlings with attached seeds had significantly lower photosynthetic rates than seedlings with removed seeds. Despite temporal variation in photosynthesis per unit leaf area, seedlings with attached seeds always had significantly greater leaf area than seedlings with removed seeds, resulting in significantly higher total plant photosynthesis at all three time periods. The delta13C values of both the leaves and roots were more similar to that of the seed for seedlings with attached seeds than for seedlings with removed seeds, however, seed removal and seed size strongly affected root delta13C. This study demonstrates that seed reserves have important effects on the early growth, physiology, and delta13C of L. densiflora seedlings. PMID- 15338416 TI - Dead wood biomass and turnover time, measured by radiocarbon, along a subalpine elevation gradient. AB - Dead wood biomass can be a substantial fraction of stored carbon in forest ecosystems, and coarse woody debris (CWD) decay rates may be sensitive to climate warming. We used an elevation gradient in Colorado Rocky Mountain subalpine forest to examine climate and species effects on dead wood biomass, and on CWD decay rate. Using a new radiocarbon approach, we determined that the turnover time of lodgepole pine CWD (340+/-130 years) was roughly half as long in a site with 2.5-3 degrees C warmer air temperature, as that of pine (630+/-400 years) or Engelmann spruce CWD (800+/-960 and 650+/-410 years) in cooler sites. Across all sites and both species, CWD age ranged from 2 to 600 years, and turnover time was 580+/-180 years. Total standing and fallen dead wood biomass ranged from 4.7+/ 0.2 to 54+/-1 Mg ha(-1), and from 2.8 to 60% of aboveground live tree biomass. Dead wood biomass increased 75 kg ha(-1) per meter gain in elevation and decreased 13 Mg ha(-1) for every degree C increase in mean air temperature. Differences in biomass and decay rates along the elevation gradient suggest that climate warming will lead to a loss of dead wood carbon from subalpine forest. PMID- 15338417 TI - Pollinator community structure and sources of spatial variation in plant- pollinator interactions in Clarkia xantiana ssp. xantiana. AB - The structure of diverse floral visitor assemblages and the nature of spatial variation in plant-pollinator interactions have important consequences for floral evolution and reproductive interactions among pollinator-sharing plant species. In this study, I use surveys of floral visitor communities across the geographic range of Clarkia xantiana ssp. xantiana (hereafter C. x. xantiana) (Onagraceae) to examine the structure of visitor communities, the specificity of the pollination system, and the role of variation in the abiotic vs. biotic environment in contributing to spatial variation in pollinator abundance and community composition. Although the assemblage of bee visitors to C. x. xantiana is very diverse (49 species), few were regular visitors and likely to act as pollinators. Seventy-four percent of visitor species accounted for only 11% of total visitor abundance and 69% were collected in three or fewer plant populations (of ten). Of the few reliable visitors, Clarkia pollen specialist bees were the most frequent visitors, carried more Clarkia pollen compared to generalist foragers, and were less likely to harbor foreign pollen. Overall, the core group of pollinators was obscured by high numbers of incidental visitors that are unlikely to contribute to pollination. In a geographic context, the composition of specialist pollinator assemblages varied considerably along the abiotic gradient spanning the subspecies' range. However, the overall abundance of specialist pollinators in plant populations was not influenced by the broad scale abiotic gradient but strongly affected by local plant community associations. C. x. xantiana populations sympatric with pollinator-sharing congeners were visited twice as often by specialists compared to populations occurring alone. These positive indirect interactions among plant species may promote population persistence and species coexistence by enhancing individual reproductive success. PMID- 15338418 TI - Auto-SENSE view-sharing cine cardiac imaging. AB - Cardiac MR cine imaging during breath hold is a compromise between spatial and temporal resolution and duration of breath hold. Especially for sick patients who have problems holding their breath, a short acquisition time is mandatory for all sequences. A combination of Auto-SENSE parallel imaging and view-sharing was implemented for fast cine imaging of the human heart and applied to healthy volunteers. Compared to conventional Fourier imaging, data acquisition could be accelerated by a factor of 3.6. Neither a pre-scan nor additional lines in k space are required to generate the sensitivity maps in Auto-SENSE. PMID- 15338419 TI - Intestinal parasites and bacteria of mountain gorillas (Gorilla beringei beringei) in Bwindi Impenetrable National Park, Uganda. AB - A survey in 1994 examined intestinal helminths and bacterial flora of mountain gorillas (Gorilla beringei beringei) in Bwindi Impenetrable National Park, Uganda. Parasites and bacteria were identified to genus in the feces of two groups of tourist-habituated and one group of non-tourist-habituated mountain gorillas. Eggs were identified as those of an anoplocephalid cestode, and nematode eggs representative of the genera: Trichuris, Ascaris, Oesophagostomum, Strongyloides, and Trichostrongylus. This is the first report of Ascaris lumbricoides-like eggs in mountain gorillas. Fecal samples (n=76) from all groups contained helminth eggs, with strongyle eggs and anoplocephalid eggs being the most common. Salmonella and Campylobacter were found in both gorilla groups. Regular long-term non-invasive fecal monitoring of the populations of mountain gorillas is essential for the prevention and identification of potential health threats by intestinal parasites and bacteria in this highly endangered subspecies. PMID- 15338420 TI - Bacterial contaminants of fuel ethanol production. AB - Bacterial contamination is an ongoing problem for commercial fuel ethanol production facilities. Both chronic and acute infections are of concern, due to the fact that bacteria compete with the ethanol-producing yeast for sugar substrates and micronutrients. Lactic acid levels often rise during bouts of contamination, suggesting that the most common contaminants are lactic acid bacteria. However, quantitative surveys of commercial corn-based fuel ethanol facilities are lacking. For this study, samples were collected from one wet mill and two dry grind fuel ethanol facilities over a 9 month period at strategic time points and locations along the production lines, and bacterial contaminants were isolated and identified. Contamination in the wet mill facility consistently reached 10(6) bacteria/ml. Titers from dry grind facilities were more variable but often reached 10(8)/ml. Antibiotics were not used in the wet mill operation. One dry grind facility added antibiotic to the yeast propagation tank only, while the second facility dosed the fermentation with antibiotic every 4 h. Neither dosing procedure appeared to reliably reduce overall contamination, although the second facility showed less diversity among contaminants. Lactobacillus species were the most abundant isolates from all three plants, averaging 51, 38, and 77% of total isolates from the wet mill and the first and second dry grind facilities, respectively. Although populations varied over time, individual facilities tended to exhibit characteristic bacterial profiles, suggesting the occurrence of persistent endemic infections. PMID- 15338421 TI - Molecular chaperones facilitate the soluble expression of N-acyl-D-amino acid amidohydrolases in Escherichia coli. AB - The overproduction of D-aminoacylase ( D-ANase, 233.8 U/mg), N-acyl-D-glutamate amidohydrolase (D-AGase, 38.1 U/mg) or N-acyl-D-aspartate amidohydrolase (D AAase, 6.2 U/mg) in Escherichia coli is accompanied by aggregation of the overproduced protein. To facilitate the expression of active enzymes, the molecular chaperones GroEL-GroES (GroELS), DnaK-DnaJ-GrpE (DnaKJE), trigger factor (TF), GroELS and DnaKJE or GroELS and TF were coexpressed with the enzymes. D-ANase (313.3 U/mg) and D-AGase (95.8 U/mg) were overproduced in an active form at levels 1.3- and 1.8-fold higher, respectively, upon co-expression of GroELS and TF. An E. coli strain expressing the D-AAase gene simultaneously with the TF gene exhibited a 4.3-fold enhancement in d-AAase activity (32.0 U/mg) compared with control E. coli expressing the D-AAase gene alone. PMID- 15338422 TI - Adaptive response of yeasts to furfural and 5-hydroxymethylfurfural and new chemical evidence for HMF conversion to 2,5-bis-hydroxymethylfuran. AB - Renewable lignocellulosic materials are attractive low-cost feedstocks for bioethanol production. Furfural and 5-hydroxymethylfurfural (HMF) are among the most potent inhibitory compounds generated from acid hydrolysis of lignocelluloses to simple sugars for fermentation. In Saccharomyces cerevisiae ATCC 211239 and NRRL Y-12632 and Pichia stipitis NRRL Y-7124, furfural and HMF inhibition were determined to be dose-dependent at concentrations from 10 to 120 mM. The yeast strains were more sensitive to inhibition by furfural than HMF at the same concentration, while combined treatment of furfural and HMF synergistically suppressed cell growth. A metabolite transformed from HMF by strain NRRL Y-12632 was isolated from the culture supernatant, and conclusively identified as 2,5-bis-hydroxymethylfuran, a previously postulated HMF alcohol, with a composition of C6H8O3 and a molecular weight of 128. It is proposed that, in the presence of HMF, the yeast reduces the aldehyde group on the furan ring of HMF into an alcohol, in a similar manner as for furfural. The accumulation of this biotransformed metabolite may be less toxic to yeast cultures than HMF, as evidenced by the rapid yeast fermentation and growth rates associated with HMF conversion. The ability of yeasts to adapt to and transform furfural and HMF offers the potential for in situ detoxification of these inhibitors and suggests a genetic basis for further development of highly tolerant strains for biofuel production. PMID- 15338423 TI - Removal of carbonaceous and nitrogenous pollutants from a synthetic wastewater using a membrane-coupled bioreactor. AB - Two modified Ludzack-Ettinger (MLE)-type membrane-coupled bioreactors (MBRs) were investigated in this study for the purpose of removing both nitrogenous and carbonaceous pollutants from a synthetic wastewater. During the first MBR experiment, removal efficiencies were high (>90%) for chemical oxygen demand (COD) and ammonia, but total nitrogenous pollutant removal efficiency was poor (approximately 25%). Bacterial community analysis of ammonia oxidizing bacteria (AOB) by a nested PCR-DGGE approach detected two Nitrosomonas-like populations and one Nitrosospira-like population. During the initial portion of the second MBR experiment, COD and ammonia removal efficiencies were similar to the first MBR experiment until the COD of the influent wastewater was increased to provide additional electron donors to support denitrification. Total nitrogen removal efficiencies eventually exceeded 90%, with a hydraulic residence time (HRT) of 24 h and a recirculation ratio of 8. When the HRT of the MBR experiment was decreased to 12 h, however, ammonia removal efficiency was adversely affected. A subsequent increase in the HRT to 18 h helped improve removal efficiencies for both ammonia (>85%) and total nitrogenous compounds (approximately 70%). Our research demonstrates that MBRs can be effectively designed to remove both carbonaceous and nitrogenous pollutants. The ability of the microbial community to switch between anoxic (denitrifying) and oxic (nitrifying) conditions, however, represents a critical process constraint for the application of MLE-type MBR systems, such that little benefit is gained compared to conventional designs. PMID- 15338424 TI - A new bioassay method for quantitative analysis of tetracyclines. AB - A new method is described for quantitative analysis of tetracycline, based on the decrease in external pH of bacterial suspensions after the addition of a glucose pulse. The decrease in external pH of these suspensions was inversely proportional to the concentration of tetracycline. The correlation coefficient of standard response lines derived from the bioassay was 0.99. Tetracycline potency was determined in six tetracycline HCl samples by the sugar pulse bioassay and a turbidimetric method. The turbidimetric assay result varied from the glucose pulse data by no more than 7 and 3% at 3 and 7 min, respectively. The procedure is rapid, precise and quantitative, and requires minimal preparation and use of media, with savings in laboratory resources and time. PMID- 15338425 TI - Genetic diversity of fragmented populations of Polygala reinii (Polygalaceae), a perennial herb endemic to Japan. AB - Genetic variation at 10 allozyme loci was analyzed in 14 populations of Polygala reinii (Polygalaceae), a perennial herb endemic to central Honshu, Japan, with a fragmented geographical distribution. The levels of genetic variation within species ( P=80.0, A=3.10, H(E)=0.303) and within populations ( P=42.1, A=1.61, H(E)=0.163) were considerably higher than the mean for other endemic plants or short-lived perennial herbs. Genetic differentiation among populations was also high ( G(ST)=0.404). The genetic distance phenogram tended to show a clustering of the populations reflecting the fragmentation of the species range. A principal component analysis revealed the same tendency, as well as three groupings of populations in the Tokai district, on the Kii Peninsula and in the northern Kinki district. A negative correlation was obtained between the levels of gene flow and geographical distance among the populations ( r=-0.745, P<0.0001). These results indicated limited gene flow among populations in P. reinii, presumably due to the geographical isolation accompanying the fragmented distribution. On the other hand, the geographical differentiation between the Japan Sea and Pacific Ocean sides was found in P. reinii, suggesting the influence of postglacial migration on the establishment of the genetic structure of this species. PMID- 15338426 TI - gamma-Tubulin, microtubule arrays, and quadripolarity during sporogenesis in the hepatic Aneura pinguis (Metzgeriales). AB - This is the first report on the organization of a quadripolar microtubule system (QMS) in polyplastidic meiosis of a hepatic with polar organizers (POs). Unlike the monoplastidic sporocytes of mosses and hornworts, in which meiotic quadripolarity can be traced to plastid division and migration, sporocytes of Aneura pinguis are polyplastidic and tetrahedrally lobed before the QMS is organized. Whereas the QMS in mosses and hornworts is plastid-based, the QMS of A. pinguis is focused at four POs where gamma tubulin (gamma-tubulin) is concentrated. An aster of microtubules emanates from each PO centered in the four cytoplasmic lobes and the opposing radial microtubules interact to form the QMS that envelops the nucleus. A functionally bipolar spindle is gradually formed as the four poles converge in pairs on either side of opposite cleavage furrows. The resulting spindle remains quadripolar. Although gamma-tubulin is most concentrated in the deeply concave poles straddling cleavage furrows, it also extends into the spindle itself. Telophase groups of chromosomes curve around the polar cleavage furrows and a phragmoplast that originates in the interzonal region guides a cell plate that extends to the equatorial cleavage furrows. Discrete POs are reformed at opposite tips of the elongated dyad nuclei in prophase II and microtubules radiating from them give rise to the spindles of second meiosis. Spindles remain sharply focused and gamma-tubulin extends into distal portions of the spindle. Interzonal phragmoplasts that expand to join with pre-established cleavage furrows mediate cytokinesis resulting in a tetrad of spores. Each young tetrad member has a radial microtubule system emanating from the nucleus. PMID- 15338427 TI - A system in which anthocyanin synthesis is induced in regenerated torenia shoots. AB - A system in which anthocyanin synthesis can be induced under defined conditions was established in regenerated torenia shoots. Leaf discs prepared from torenia plantlets grown under sterile conditions were placed on solidified half-strength MS medium containing 3% sucrose and 4.4x10(-6) M benzyladenine (BA) and cultured under 16 h light/8 h dark (standard light) conditions for 10 days, then in the dark for a further 10 days. The discs were transferred to medium containing 7% sucrose without BA and cultured under standard light conditions. Six days after transfer, anthocyanin synthesis started in the regenerated shoots, and thereafter, anthocyanin accumulation increased while chlorophyll content decreased. Experiments in which either the timing of illumination was altered or shoots were retransferred to medium containing 1.5% sucrose or other sugars as well as sucrose indicated that both osmotic stress and light are required to induce anthocyanin synthesis. Once anthocyanin synthesis was induced in the torenia shoots 6 days after transfer, the shoots were fated to the synthesis of anthocyanins and the degradation of chlorophylls, and could not revert to the developmental pathway of shoot regeneration. This system may provide a good model for the investigation of the mechanisms underlying the induction of anthocyanin synthesis. PMID- 15338428 TI - Seed morphology and variation in the genus Pachycereus (Cactaceae). AB - Seeds of 13 Pachycereus species and two Stenocereus species that have been suggested as closely related were examined with the scanning electron microscope. Quantitative features were evaluated using multivariate analysis in order to identify characters that distinguish them. Several species groups were recognized on the basis of 16 qualitative characters. All species studied are keeled. Stenocereus aragonii and S. eichlamii share with most Pachycereus species large size, glossy appearance, and a flat relief on periclinal cells in the lateral region. Pachycereus gatesii and P. schottii are unique in having the smallest seeds and a deeply impressed hilum-micropylar region. P. hollianus does not exhibit micro-relief on periclinal walls in the lateral region, and P. fulviceps has no expanded testa border. Multivariate analysis showed that four characters, length, breadth, hilum-micropylar region length, and angle, made the greatest contribution to distinguishing among species groups. More than 80% of P. fulviceps, P. hollianus, P. tepamo, P. weberi, and S. eichlamii seeds could be classified correctly using four seed features and the percentage was even higher using just two or three features for P. gatesii, P. grandis, P. militaris, P. pringlei, and P. schottii. Testa appearance, testa cell-pattern, and position relative to the rim of the hilum-micropylar region were found to be potentially informative and should be combined with other sources of data in future phylogenetic analyses. PMID- 15338429 TI - Resurrection of an ancestral gene: functional and evolutionary analyses of the Ngrol genes transferred from Agrobacterium to Nicotiana. AB - The Ng rol genes, which have high similarity in sequence to the rol genes of Agrobacterium rhizogenes, are present in the genome of untransformed plants of Nicotiana glauca. It is thought that bacterial infection resulted in the transfer of the Ng rol genes to plants early in the evolution of the genus Nicotiana, since several species in this genus contain rol-like sequences but others do not. Plants transformed with the bacterial rol genes exhibit various developmental and morphological changes. The presence of rol-like sequences in plant genomes is therefore thought to have contributed to the evolution of Nicotiana species. This paper focuses on studies of the Ng rol genes in present-day plants and during the evolution of the genus Nicotiana. The functional sequences of several Ng rol genes may have been conserved after their ancient introduction from a bacterium to the plant. Resurrection of an ancestral function of one of the Ng rol genes, as examined by physiological and evolutionary analyses, is also described. The origin of the Ng rol genes is then considered, based on results of molecular phylogenetic analyses. The effects of the horizontal transfer of the Ng rol genes and mutations in the genes are discussed on the plants of the genus Nicotiana during evolution. PMID- 15338430 TI - Cytolethal distending toxins. AB - The cytolethal distending toxins (CDTs) constitute the most recently discovered family of bacterial protein toxins. CDTs are unique among bacterial toxins as they have the ability to induce DNA double strand breaks (DSBs) in both proliferating and nonproliferating cells, thereby causing irreversible cell cycle arrest or death of the target cells. CDTs are encoded by three linked genes ( cdtA, cdtB and cdtC) which have been identified among a variety of Gram-negative pathogenic bacteria. All three of these gene products are required to constitute the fully active holotoxin, and this is in agreement with the recently determined crystal structure of CDT. The CdtB component has functional homology with mammalian deoxyribonuclease I (DNase I). Mutation of the conserved sites necessary for this catalytic activity prevents the induction of DSBs as well as all subsequent intoxication responses of target cells. CDT is endocytosed via clathrin-coated pits and requires an intact Golgi complex to exert the cytotoxic activity. Several issues remain to be elucidated regarding CDT biology, such as the detailed function(s) of the CdtA and CdtC subunits, the identity of the cell surface receptor(s) for CDT, the final steps in the cellular internalization pathway, and a molecular understanding of how CDT interacts with DNA. Moreover, the role of CDTs in the pathogenesis of diseases still remains unclear. PMID- 15338431 TI - The foam/emulsion analogy in structure and drainage. AB - The often quoted analogy between foams and emulsions is experimentally tested by studying properties after settling and under forced drainage of oil-in-water emulsions of drop size similar as for bubbles generally used in foam experiments. Observations with regard to structure, water fraction and drainage wave properties confirm the expected similarity in the low flow rate range. However, while for foams a convective circulation on the scale of the container sets in for values of water fraction exceeding about 0.2, no such convection is found in emulsions. Here instabilities are only encountered at water fractions of about 0.4, close to the void fraction of random packings of spheres. These take on the form of descending pulses of increased water fraction and lead to the transition from a frozen to a locally agitated structure. PMID- 15338432 TI - Mechanical properties of mono-domain side chain nematic elastomers. AB - We investigate the behavior of the shear rigidity modulus G = G' + iG" of three mono-domain side chain liquid-crystalline elastomers composed of side chain polysiloxanes cross-linked by either flexible or rigid cross-linkers. The measurements were taken in a frequency domain ranging from approximately 0.02 Hz to approximately 10(4) Hz applying the shear in a plane perpendicular to or containing the director. The measurements as a function of temperature show an anisotropy of G' which appears around T(NI), when decreasing the temperature, and which is due to the expected lowering of G'(//) coming from the coupling between the shear and the director. The measurements as a function of frequency show that G has two components for both geometries, in both the isotropic phase and in the nematic phase around the phase transition. One reflects the network behavior in its hydrodynamic regime ( G' is constant and G'' is approximately f, where f is the frequency), the other which appears at higher frequencies is characterized by a scaling law behavior (G' is approximately G" is approximately f(0.5)) of the Rouse type. We discuss the results in the framework of available theories and show that the three elastomers present a non-soft behavior, even for the elastomer for which the contrary was claimed, and that there is no separation of time scales between the director and the network. We also present data on a poly domain sample and a non-mesomorphic one which complement these results. PMID- 15338433 TI - Transients in sheared granular matter. AB - As dense granular materials are sheared, a shear band and an anisotropic force network form. The approach to steady-state behavior depends on the history of the packing and the existing force and contact network. We present experiments on shearing of dense granular matter in a 2D Couette geometry in which we probe the history and evolution of shear bands by measuring particle trajectories and stresses during transients. We find that when shearing is stopped and restarted in the same direction, steady-state behavior is immediately reached, in agreement with the typical assumption that the system is quasistatic. Although some relaxation of the force network is observed when shearing is stopped, quasistatic behavior is maintained because the contact network remains essentially unchanged. When the direction of shear is reversed, a transient occurs in which stresses initially decrease, changes in the force network reach further into the bulk, and particles far from the wheel become more mobile. This occurs because the force network is fragile to changes transverse to the force network established under previous shear; particles must rearrange before becoming jammed again, thereby providing resistance to shear in the reversed direction. The strong force network is re-established after displacing the shearing surface approximately equal 3d, where d is the mean grain diameter. Steady-state velocity profiles are reached after a shear of < or approximately equal 30 d. Particles immediately outside of the shear band move on average less than 1 diameter before becoming jammed again. We also examine particle rotation during this transient and find that mean particle spin decreases during the transient, which is related to the fact that grains are not interlocked as strongly. PMID- 15338436 TI - Rheological properties of a highly confined film of a lyotropic lamellar phase. AB - We investigated the rheological properties of a thin film of a lyotropic lamellar phase with a Dynamic Surface Forces Apparatus. The minimum thickness of the film is varied between one to several tens of layers by confining the materials between solid surfaces. The rheometric properties are measured with the application of a small harmonic compression. These properties depend clearly on the smectic order of the material. Whole mechanical properties may be easily described by taking into account interactions between membranes and motion of the dislocation line loops. In particular, it is shown that at the dynamical frequencies investigated in this study, the solvent flows between membranes which remain undeformed. Consequences and perspectives of this study will be discussed. PMID- 15338437 TI - Structure of hydrophobically and hydrogen-bonded complexes between amphiphilic copolymer and polyacid in water. AB - We observed by SANS and NMR the structure of intermolecular complexes formed through hydrogen bonding and hydrophobic interactions between a polyacid and a neutral copolymer surfactant (PEO-PPO-PEO). The polyacid is perdeuterated and the contrast variation method enables us to measure separately the structure factor of each component in the complex and the cross structure factor as a function of the pH and the temperature. The evolution of NMR spectra and relaxation times with pH and temperature give a complementary insight into the inner structure of the aggregates. The interaction between the aggregates and the aggregation number of polyacid chains are governed by electrostatic effects; they depend on the polyacid charge and are insensitive to temperature. The number of copolymer chains which results from the cooperative action of hydrogen bonding and hydrophobic interactions is sensitive to both pH and temperature. The complexation preserves the micellar core-corona structure of the copolymer and shrinks the polyacid chains which adopt a compact structure. The non-dissociated polyacid sequences are bound to the PPO part of the copolymer forming the hydrophobic core of the complex, whereas the ionized polyacid sequences form with the PEO a stabilizing hydrated corona. PMID- 15338438 TI - Hemifusion and fusion of giant vesicles induced by reduction of inter-membrane distance. AB - Proteins involved in membrane fusion, such as SNARE or influenza virus hemagglutinin, share the common function of pulling together opposing membranes in closer contact. The reduction of inter-membrane distance can be sufficient to induce a lipid transition phase and thus fusion. We have used functionalized lipids bearing DNA bases as head groups incorporated into giant unilamellar vesicles in order to reproduce the reduction of distance between membranes and to trigger fusion in a model system. In our experiments, two vesicles were isolated and brought into adhesion by the mean of micromanipulation; their evolution was monitored by fluorescence microscopy. Actual fusion only occurred in about 5% of the experiments. In most cases, a state of "hemifusion" is observed and quantified. In this state, the outer leaflets of both vesicles' bilayers merged whereas the inner leaflets and the aqueous inner contents remained independent. The kinetics of the lipid probes redistribution is in good agreement with a diffusion model in which lipids freely diffuse at the circumference of the contact zone between the two vesicles. The minimal density of bridging structures, such as stalks, necessary to explain this redistribution kinetics can be estimated. PMID- 15338439 TI - On the polarization of chiral main-chain liquid-crystalline elastomers. AB - A mechanism of developing a polarization in a chiral main-chain liquid crystalline polymer by aligning the dipoles of the monomers is explored. It is shown that the polarization of a pure liquid crystal elastomer is zero in equilibrium due to rotation of the director in the elastomer. A constraint or specific non-ideality in the elastomer is required to prevent this relaxation of the director in order to realize a non-zero polarization. Three methods that circumvent this result are explored. We consider the effect of an oscillating shear, the pinning effect of the layers in a smectic-A composed of chiral smectogens and a binary mixture of chiral main chains and non-chiral side chains. Each of these methods is shown to produce a polarization, which is much larger than that produced in a piezoelectric alpha-quartz crystal per unit stress. PMID- 15338440 TI - Dynamics of concentrated colloidal suspensions during drying--aging, rejuvenation and overaging. AB - We report on the slow dynamics of concentrated colloidal suspensions during drying and rewetting under conditions of reversible concentration changes without coalescence or aggregation. We used multispeckle diffusing-wave spectroscopy to monitor the slow dynamics of particles. We found that the alpha relaxation of the suspensions exhibits successively slowing-down, acceleration and a stationary regime during drying at constant rates. Under rewetting conditions, we observed slowing-down and a stationary regime. The characteristic time of the stationary regime is inversely proportional to the rate of concentration change and identical for both drying and rewetting. We explain these regimes as aging (overaging), rejuvenation and plastic flow of the suspensions induced by a deviatoric stress (a combination of compressive and elongational stresses) which is induced by the uniaxial compressive strain generated by evaporation. PMID- 15338441 TI - Tilt modulus of a lipid monolayer. AB - In addition to the familiar bending and stretching deformations, lipid monolayers and bilayers in their disordered state are often subjected to tilt deformations, occurring for instance in structural rearrangements accompanying membrane fusion, or upon insertion of "oblique" hydrophobic proteins into lipid bilayers. We study the elastic response of a flat lipid monolayer to a tilt deformation, using the spatial and conformational average of the chain end-to-end vector from the membrane normal to define a macroscopic membrane tilt. The physical origin and magnitude of the corresponding tilt modulus kappat is analyzed using two complementary theoretical approaches. The first is a phenomenological model showing that the tilt and bending deformations are decoupled and the effects of inter-chain correlations on the tilt modulus is small. The second is based on a molecular-level mean-field theory of chain packing, enabling numerical evaluation of the tilt modulus for realistic, multi-conformation, chain models. Both approaches reveal that the tilt modulus involves two major contributions. The first is elastic in origin, arising from the stretching of the hydrocarbon chains upon a tilt deformation and reflecting the loss of chain conformational freedom associated with chain stretching. The second, purely entropic, contribution results from the constraints imposed by a tilt deformation on the fluctuations of chain director orientations. Using the chain-packing theory we compute the two contributions numerically as a function of the cross-sectional area per chain. The elastic and entropic terms are shown to dominate the value of kappat for small and large areas per chain, respectively. For typical cross-sectional areas of lipid chains in biological membranes they are of comparable magnitude, yielding kappat approximately 0.2kBT/A2. PMID- 15338442 TI - Visualization of large-scale correlations in gene expressions. AB - Large-scale expression data are today measured for several thousands of genes simultaneously. Furthermore, most genes are being categorized according to their properties. This development has been followed by an exploration of theoretical tools to integrate these diverse data types. A key problem is the large noise level in the data. Here, we investigate ways to extract the remaining signals within these noisy data sets. We find large-scale correlations within data from Saccharomyces cerevisiae with respect to properties of the encoded proteins. These correlations are visualized in a way that is robust to the underlying noise in the measurement of the individual gene expressions. In particular, for S. cerevisiae we observe that the proteins corresponding to the 400 highest expressed genes typically are localized to the cytoplasm. These most expressed genes are not essential for cell survival. PMID- 15338443 TI - Apoptosis of gastric cancer cell line MKN45 by photodynamic treatment with photofrin. AB - The aim of this study was to investigate the mechanism of cell death by photodynamic therapy (PDT) in the gastric cancer cell line MKN45 with focus on the mechanism of apoptosis. Gastric cancer cells (MKN45) were incubated with Photofrin for up to 24 h before exposure to He-Cd laser (441 nm, 1 J/cm2). Cell viability was assessed by the methyl-tetra-zolium assay after exposure to light. A 95% cell death (LD95) was measured with 10 microg/ml of Photofrin. DNA ladder formation and chromatin condensation were seen within 60 min. Caspase-3-like and caspase-9-like activities increased from 15 min after exposure to light. Reduction of rhodamine 123 uptake started at 30 min. Caspase-inhibitor VAD-fmk (10 mM) inhibited apoptosis, but did not influence cell viability. In conclusion, Photofrin-mediated PDT in the gastric cancer cell line MKN45 induces apoptosis within 60 min, and mitochondrial damage is likely as the first event of apoptosis. PMID- 15338444 TI - Operator error is the key factor contributing to medical laser accidents. AB - Useful lessons can often be learned from studying laser accidents. Although most countries require such accidents to be reported to the relevant authority, often such information is not accessible to laser users. Consequently, it was decided to collect detailed accounts of incidents from the British Medical Laser Association's Internet mailing list. Twelve incidents were reported by e-mail after a request for information sent to the BMLA mailing list in 2003. Although these are of a diverse nature, operator error accounted for 67% of the cases. This factor should be recognized when developing laser safety programs. PMID- 15338446 TI - Dogs (Canis lupus familiaris) fail to show understanding of means-end connections in a string-pulling task. AB - Domestic dogs (Canis lupus familiaris) were tested in four experiments for their understanding of means-end connections. In each of the experiments, the dogs attempted to retrieve a food treat that could be seen behind a barrier and which was connected, via string, to a within-reach wooden block. In the experiments, either one or two strings were present, but the treat was attached only to one string. Successful retrieval of the treat required the animals to pull the appropriate string (either by pawing or by grasping the wooden block in their jaws) until the treat emerged from under the barrier. The results showed that the dogs were successful if the treat was in a perpendicular line to the barrier, i.e. straight ahead, but not when the string was at an angle: in the latter condition, the typical response was a proximity error in that the dogs pawed or mouthed at a location closest in line to the treat. When two strings that crossed were present, the dogs tended to pull on the wrong string. The combined results from the experiments show that, although dogs can learn to pull on a string to obtain food, they do not spontaneously understand means-end connections involving strings. PMID- 15338445 TI - A case of cellulitis complicating Campylobacter jejuni subspecies jejuni bacteremia and review of the literature. AB - Infection with Campylobacter species is a predominant cause of food-borne gastroenteritis in the industrialized world. Bacteremia is detected in <1% of patients with diarrhea, mainly in immunocompromised hosts or those in the extremes of age. Reported here is the case of a 78-year-old, immunocompromised male patient with Campylobacter jejuni subsp. jejuni bacteremia complicated by cellulitis. The infection was characterized by a protracted course with several recurrences and refractoriness to multiple antibiotic regimens, responding only to a prolonged course of meropenem treatment. The frequency of cellulitis as reflected in previously reported series of Campylobacter bacteremia and the clinical characteristics of this difficult-to-treat infection are reviewed. PMID- 15338447 TI - Relationship between high-resolution computed tomography findings and the Stoke index in patients with rheumatoid arthritis. AB - The aim of this study was to evaluate the relationship between high-resolution computed tomography (HRCT) findings and the Stoke index (SI) in patients with rheumatoid arthritis (RA). Forty RA patients (31 women, 9 men) were evaluated. All patients fulfilled the criteria proposed by the American College of Rheumatology. Clinical evaluation, haematological data, chest radiography, pulmonary function tests (PFTs) and HRCT were obtained in all patients. The SI was used to assess disease activity. In 17 (42.5%) patients, there were no signs of pulmonary involvement on HRCT; 23 (57.5%) of 40 patients had abnormal HRCT findings. Of 23 patients with HRCT abnormalities, six (two male, four female) had respiratory symptoms, four (one male, three female) had abnormalities on chest radiography and five (all female) had abnormalities on PFTs. There was no relationship between pulmonary changes observed on HRCT, clinical and laboratory disease activity parameters, chest X-ray and PFTs. There was no difference in the mean SI between patients included in the HRCT (+) and those included in the HRCT (-) groups. No significant correlations between the HRCT and the SI were seen. The main findings of this study are that HRCT can give useful information on RA associated lung changes and that there was no relationship between the SI and the HRCT findings of patients with RA. PMID- 15338448 TI - Relationship between self-efficacy and exercise duration in patients with ankylosing spondylitis. PMID- 15338449 TI - A case of dermatomyositis associated with papillary cancer of the thyroid gland. PMID- 15338450 TI - Unusual association of systemic sclerosis and ankylosing spondylitis. PMID- 15338451 TI - Levels of soluble E-selectin in patients with active Behcet's disease. AB - Behcet's disease is a systemic vasculitis of unknown aetiology. Endothelial cell injury plays an important role in the pathogenesis and immunopathology of Behcet's disease. E-selectin is expressed by activated endothelial cells. Because the selectin adhesion molecules are shed from activated cells, soluble forms of these proteins can be used as activation markers of endothelium (E-selectin). The pathogenesis of Behcet's disease (BD) is closely related to endothelial cells, leucocyte functions and immunity. The aim of this study was to investigate circulating E-selectin adhesion molecules, which are known to play a significant part in the immune response especially by regulating interaction of the leucocytes with endothelium in BD. Plasma E-selectin concentrations were evaluated in 23 patients with BD and 20 healthy control subjects. The disease activity was evaluated by clinical manifestations (oral aphthous ulcer, genital ulceration, positive pathergy test, skin lesions, eye involvement, thrombophlebitis and arthritis) and by laboratory investigations [erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP)]. The patients were newly or previously diagnosed cases not taking any drug for BD. Levels of E-selectin were measured with commercially available sandwich enzyme-linked immunosorbent assay (ELISA) kits using human sE-selectin (cat. no: BMS 205). Plasma E-selectin concentrations of patients and controls were compared with the Mann-Whitney U test. Statistical significance was assigned to p values lower than 0.05. Serum levels (mean+/-SD) of soluble E-selectin (sE-selectin) were significantly higher in 23 patients with BD than in 20 healthy controls (53.2+/-18.2 ng/ml vs 33.8+/ 7.5 ng/ml, p<0.0001). A statistically significant positive correlation was observed between sE-selectin levels and CRP and ESR in patients with BD (r=0.78, p<0.001 and r=0.56, p<0.01, respectively). Increases in the E-selectin in BD may be a direct consequence of the leucocyte and endothelium activations observed during the disease process. The noninvasive investigations can be used as biochemical markers for inflammation. This may provide additional information regarding disease activity along with the traditional indices such as ESR and CRP. PMID- 15338452 TI - Acute respiratory distress syndrome due to systemic lupus erythematosus with hemophagocytic syndrome: an autopsy report. AB - This report concerns a patient with systemic lupus erythematosus (SLE) who died of acute respiratory distress syndrome (ARDS) 1 day after the onset of pulmonary symptoms. Autopsy demonstrated severe hemophagocytosis in the bone marrow and histopathology indicating a marked increase in vascular permeability in both lungs and kidneys. In this patient, active SLE and associated hemophagocytic syndrome may have induced an increase in the production of inflammatory cytokines, which immediately induced ARDS. Since fatal ARDS can occur as a life threatening complication of SLE, careful observation is necessary, particularly when there are clinical findings suggestive of associated hemophagocytic syndrome. PMID- 15338453 TI - Genetic analysis of the cardiac sodium channel gene SCN5A in Koreans with Brugada syndrome. AB - The SCN5A gene encodes the alpha subunit of the human cardiac voltage-gated sodium channel. Mutations in SCN5A are responsible for Brugada syndrome, an inherited cardiac disease that leads to idiopathic ventricular fibrillation (IVF) and sudden death. In this study, we screened nine individuals from a single family and 12 sporadic patients who were clinically diagnosed with Brugada syndrome. Using PCR-SSCP, DHPLC, and DNA sequencing analysis, we identified a novel single missense mutation associated with Brugada syndrome in the family and detected a C5607T polymorphism in Korean subjects. A single nucleotide substitution of G to A at nucleotide position 3934 changed the coding sense of exon 21 of the SCN5A from glycine to serine (G1262S) in segment 2 of domain III (DIII-S2). Four individuals in the family carried the identical mutation in the SCN5A gene, but none of the 12 sporadic patients did. This mutation was not found in 150 unrelated normal individuals. This finding is the first report of a novel mutation in SCN5A associated with Brugada syndrome in Koreans. PMID- 15338455 TI - Comparative analysis of trehalose production by Debaryomyces hansenii and Saccharomyces cerevisiae under saline stress. AB - The comparative analysis of growth, intracellular content of Na+ and K+, and the production of trehalose in the halophilic Debaryomyces hansenii and Saccharomyces cerevisiae were determined under saline stress. The yeast species were studied based on their ability to grow in the absence or presence of 0.6 or 1.0 M NaCl and KCl. D. hansenii strains grew better and accumulated more Na+ than S. cerevisiae under saline stress (0.6 and 1.0 M of NaCl), compared to S. cerevisiae strains under similar conditions. By two methods, we found that D. hansenii showed a higher production of trehalose, compared to S. cerevisiae; S. cerevisiae active dry yeast contained more trehalose than a regular commercial strain (S. cerevisiae La Azteca) under all conditions, except when the cells were grown in the presence of 1.0 M NaCl. In our experiments, it was found that D. hansenii accumulates more glycerol than trehalose under saline stress (2.0 and 3.0 M salts). However, under moderate NaCl stress, the cells accumulated more trehalose than glycerol. We suggest that the elevated production of trehalose in D. hansenii plays a role as reserve carbohydrate, as reported for other microorganisms. PMID- 15338456 TI - The G/G genotype of a resistin single-nucleotide polymorphism at -420 increases type 2 diabetes mellitus susceptibility by inducing promoter activity through specific binding of Sp1/3. AB - Insulin resistance is a major cause of type 2 diabetes mellitus (T2DM). Resistin, an adipocyte-secreted hormone, antagonizes insulin. Transgenic mice that overexpress the resistin gene (Retn) in adipose tissue are insulin-resistant, whereas Retn (-/-) mice show lower fasting blood glucose, suggesting that the altered Retn promoter function could cause diabetes. To determine the role of RETN in human T2DM, we analyzed polymorphisms in its 5' flanking region. We found that the -420G/G genotype was associated with T2DM (397 cases and 406 controls) (P=.008; adjusted odds ratio = 1.97 [by logistic regression analysis]) and could accelerate the onset of disease by 4.9 years (P=.006 [by multiple regression analysis]). Meta-analysis of 1,888 cases and 1,648 controls confirmed this association (P=.013). Linkage disequilibrium analysis revealed that the -420G/G genotype itself was a primary variant determining T2DM susceptibility. Functionally, Sp1 and Sp3 transcription factors bound specifically to the susceptible DNA element that included -420G. Overexpression of Sp1 or Sp3 enhanced RETN promoter activity with -420G in Drosophila Schneider line 2 cells that lacked endogenous Sp family members. Consistent with these findings, fasting serum resistin levels were higher in subjects with T2DM who carried the -420G/G genotype. Therefore, the specific recognition of -420G by Sp1/3 increases RETN promoter activity, leading to enhanced serum resistin levels, thereby inducing human T2DM. PMID- 15338457 TI - No "bias" toward the null hypothesis in most conventional multipoint nonparametric linkage analyses. PMID- 15338458 TI - Conventional multipoint nonparametric linkage analysis is not necessarily inherently biased. PMID- 15338459 TI - "Bias toward the null" means reduced power. PMID- 15338460 TI - No bias in linkage analysis. PMID- 15338462 TI - Germline PHOX2B mutation in hereditary neuroblastoma. PMID- 15338463 TI - Comparative frequency of fragile-X (FMR1) and creatine transporter (SLC6A8) mutations in X-linked mental retardation. PMID- 15338465 TI - Tartrate-resistant acid phosphatase: a potential target for therapeutic gold. AB - Gold compounds are disease-modifying agents for the treatment of rheumatoid arthritis. They act on the immune system but the mechanism is not fully understood. Gold has been shown to affect antigen processing by T-cells and also reduces expression of cytokines in macrophages. Tartrate-resistant acid phosphatase (TRAP), expressed by osteoclasts, macrophages and dendritic cells is an enzyme with roles in skeletal metabolism and the immune response. TRAP is able to degrade skeletal phosphoproteins including osteopontin, identical to the T cell cytokine, Eta-1; we thus propose that TRAP regulates the Eta-1 pathway common to the immune system and skeleton. We compared the distribution of osteopontin and TRAP in sections of 18-day-old embryonic mice by immunohistochemistry. Both proteins occurred in the same locations. To determine whether gold compounds exert their effects by modification of TRAP activity, we examined the action of gold chloride and the prodrugs, aurothioglucose and aurothiomalate on the dephosphorylation of osteopontin by TRAP. Aurothioglucose and aurothiomalate had little effect on phosphatase activity; gold chloride was a potent non-competitive inhibitor (Ki < 47 x 10(-9) M). These findings indicate a possible molecular mechanism for the action of therapeutic gold and further implicate TRAP in the control of immunity. PMID- 15338466 TI - Effects of therapeutic and toxic doses of levamisole on thyroid hormones and some biochemical parameters in sheep. AB - This study was carried out to establish the effects of therapeutic and toxic doses of levamisole on thyroid hormone levels and some biochemical parameters in sheep. Twelve Akkaraman ewes were used. Levamisole was given orally at doses of 7.5 mg kg(-1) (group 1) and 40 mg kg(-1) (group 2) to the animals. Blood samples were taken from the jugular vein at 2, 4, 8, 24, 48, 96 and 144 h after the administrations. Serum thyroid hormones and some biochemical parameters were determined on these samples. When compared with the control levels, no significant changes were observed in triiodothyronine (T3) and thyroxin (T4) levels in group 1. Although levamisole was found to increase the levels of total T3, it decreased the levels of total T4 in group 2. On the other hand, free T3 and free T4 levels were not changed in either group. While serum alkaline phosphatase (ALP) activities were decreased, aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and creatinine kinase (CK) activities were increased significantly by levamisole. However, it increased the serum albumin and cholesterol levels, but decreased the inorganic phosphate levels in groups 1 and 2. On the other hand, when compared with the control levels, no significant changes were detected in serum sodium, potassium and calcium levels. In conclusion, therapeutic and toxic doses of levamisole were determined to affect thyroid metabolism and some biochemical parameters in sheep. PMID- 15338467 TI - The effects of the caffeic acid phenethyl ester (CAPE) on erythrocyte membrane damage after hind limb ischaemia-reperfusion. AB - Reactive oxygen species have been implicated in pathogenesis injury after ischaemia-reperfusion (I/R). Caffeic Acid Phenethyl Ester (CAPE), an active component of honeybee propolis extract, exhibits antioxidant and anti inflammatory properties. The aim of this study was to investigate the effects of CAPE on erythrocyte membrane damage after hind limb I/R. Rats were divided into two groups: I/R and I/R with CAPE pre-treatment. They were anaesthetized with intramuscular ketamine 100 mg kg(-1). A 4-h I/R period was performed on the right hind limb of all animals. In the CAPE-treated group, animals received CAPE 10 microm by intraperitoneal injection 1 h before the reperfusion. At the end of the reperfusion period, a midsternotomy was performed. A 5-ml blood sample was withdrawn from the ascending aorta for biochemical assays. Serum and erythrocyte membrane MDA levels were significantly lower in the CAPE-treated group when compared to the I/R group ( p = 0.001 and p<0.001, respectively). Erythrocyte membrane Na(+)-K(+) ATPases activity in the CAPE-treated group was significantly higher than the I/R group ( p<0.001). In conclusion, CAPE seems to be effective in protecting against oxidative stress. Therefore, we suggest that in order to decrease I/R injury, pre-administration of CAPE may be a promising agent for a variety of conditions associated with I/R. PMID- 15338468 TI - Effects of protein kinase C on M-phase promoting factor in early development of fertilized mouse eggs. AB - The mechanism of development of mouse fertilized eggs from the one-cell stage to the two-cell stage remains unclear to date. In the present study, we have evaluated protein kinase C (PKC) and M-phase promoting factor (MPF) kinase activity in fertilized mouse eggs treated with a PKC modulator. PKC and MPF activity have similar activity. The two subunits of MPF, p34(cdc2) and cyclin B, were shown to be included in the substrates phosphorylated by PKC in fertilized mouse eggs, while PKC modulator affected the electrophoretic mobility shift of cdc2 and cdc25C by dephosphorylation and phosphorylation. These results clearly indicate that PKC may affect the progression of the cell cycle through post translational modification of MPF activity. PMID- 15338469 TI - Protective effects of intraperitoneal vitamin C, aprotinin and melatonin administration on retinal edema during experimental uveitis in the guinea pig. AB - A considerable amount of clinical and experimental evidence exists suggesting the involvement of reactive oxygen substances (ROS) in the aetiology of uveitis. The activated phagocytic system of polymorphonuclear leucocytes in uveitis is involved in the generation of ROS. In addition to their direct free radical scavenging action, aprotinin, melatonin and vitamin C are known to protect against oedema formation and can preserve plasma membrane fluidity and free radical production. Histological changes in the retina that occur during uveitis are not well explained. The purpose of this study was to determine whether vitamin C, aprotinin and melatonin can protect the retina from damage accompanying experimental uveitis (EU). Thirty adult male guinea pigs were divided into five groups of six animals each. The first group was used as control. The right eyes of groups 2, 3, 4 and 5 received an intravitreal injection of bovine serum albumin for induction of experimental uveitis. At the same time and also on the consecutive third day, groups 3, 4 and 5 received intraperitoneal injections of vitamin C (ascorbic acid, 100 mg kg(-1) body wt), aprotinin (20,000 kIU kg(-1) body wt) and melatonin (10 mg kg(-1) body wt), respectively. The animals were killed on the sixth day. The average thickness of the retina and inner plexiform layer for each eye was measured in sagittal section near the optic nerve and expressed in microns. The thickness of the retina and inner plexiform layer in the control group was significantly (p < 0.01) lower than in the group EU as compared with the group EU plus vitamin C, group EU plus aprotinin, group EU plus melatonin (p < 0.05). The thicknesses of the retina and inner plexiform layer in group EU plus vitamin C, group EU plus aprotinin and group EU plus melatonin were significantly (p < 0.01) lower than that in the group EU. The difference in thickness of the retina and inner plexiform layer among the groups 3, 4 and 5 was not significant (p > 0.05). In conclusion, this study demonstrated that oedematous effects of EU on the retina were reduced by the administration of intraperitoneal vitamin C, aprotinin and melatonin, i.e. these antioxidants had significant protective effects on the retina of guinea pigs against oedematous damage in EU. However, the reductive effect of vitamin C on EU was greater than that of aprotinin and melatonin. The intraperitoneal vitamin C, aprotinin and melatonin supplementations may strengthen the antioxidant defence system because of decreased ROS, and these agents may play a role in treating uveitis. PMID- 15338470 TI - Genetic instability of RBM5/LUCA-15/H37 in MCF-7 breast carcinoma sublines may affect susceptibility to apoptosis. AB - The MCF-7 human breast carcinoma cell line is widely used as a model system by breast cancer researchers and cell biologists investigating apoptosis. Since its establishment 30 years ago, from a patient with metastatic breast cancer, the original MCF-7 cell population has undergone genetic drift to such an extent that numerous genetically diverse sublines now exist. For instance, it has been reported that MCF-7 cells have lost the region 3p21.3, to which the apoptosis regulatory protein and putative tumour suppressor LUCA-15 (also called RBM5 and H37) maps; however, LUCA-15 has been cloned from MCF-7 cells, and LUCA-15 expression analyses have been conducted using MCF-7 cells. To address this discrepancy, we characterized three MCF-7 sublines by Western blot, RT-PCR and finally genomic PCR analysis, and determined that one of the three had lost the LUCA-15 gene. Interestingly, loss of LUCA-15 was positively correlated with decreased susceptibility to the death-inducing ligand TNF-alpha. Subsequent overexpression of exogenous LUCA-15 was shown to enhance TNF-alpha-mediated apoptosis, suggesting that LUCA-15 may play a role in regulating the susceptibility of breast cancer cells to drug-induced apoptosis. These results not only reinforce the necessity of MCF-7 subline characterization, but provide the first evidence of an apoptotic modulatory role for LUCA-15 in a non-T cell line. PMID- 15338471 TI - Protein kinase A activity is associated with metacyclogenesis in Leishmania amazonensis. AB - Because of the importance of cell signalling processes in proliferation and differentiation, the adenylate cyclase pathway was studied, specifically the protein kinase A (PKA) in Leishmania amazonensis. The PKAs of soluble (SF) and enriched membrane fractions (MF) from infective/non-infective promastigotes and axenic amastigotes were assayed. In order to purify the PKA molecule, fractions were chromatographed on DEAE-cellulose columns and the phosphorylative activity was evaluated using [gamma(32)P]-ATP as the phosphate source. These experiments were performed in the presence of cyclic adenosine monophosphate (cAMP) and an inhibitor of PKA. Our data demonstrated that the PKA activity was significantly higher (about two times) in SF from promastigotes with a high concentration of metacyclic forms, when compared with the non-infective promastigotes, suggesting an association of this activity and the metacyclogenesis process. A discrete phosphorylative activity in axenic amastigotes was observed. As the adenylate cyclase/cAMP pathway would be involved in the parasite-host interiorization, the PKA activity may constitute a good intracellular target for studies of leishmanicidal drugs. PMID- 15338472 TI - Glucose and glutamine utilization by rat lymphocytes, monocytes and neutrophils in culture: a comparative study. AB - Glucose and glutamine utilization and production of glutamate and lactate were determined for up to 48 h in lymphocytes, monocytes and neutrophils cultured in medium rich in metabolites and vitamins. Glucose was utilized by the three cell types in culture in the following order: neutrophils > monocytes > lymphocytes, whereas lactate was produced in the order: monocytes > neutrophils > lymphocytes. The consumption of glucose followed the activity of glucose-6-phosphate dehydrogenase but it was not related to hexokinase activity. Glutamine was consumed by the three leukocyte types in culture as follows: neutrophils > lymphocytes > or = monocytes. The consumption of glutamine was not fully related to the activity of phosphate-dependent glutaminase. The production of glutamate was not remarkably different among the three cell types. For comparison, glutamine and glucose utilization and glutamate and lactate production were also evaluated using 1-h incubated leukocytes. Under this condition, only glucose or glutamine was added to the medium. Glucose was utilized as follows: neutrophils > monocytes > lymphocytes, whereas lactate was produced in the following order: monocytes > or = neutrophils > lymphocytes. Glutamine was consumed as follows: neutrophils > lymphocytes > monocytes, whereas glutamate was produced as follows: neutrophils > or = monocytes = lymphocytes. The ratio of the amount of glucose/glutamine consumed by 1-h incubated cells was 0.5 for neutrophils, 1.5 for monocytes, and 0.3 for lymphocytes. However, the three cell types cultured for 48 h utilized glucose to a much higher degree than glutamine. The ratio of the amount of glucose/glutamine utilized by the cultured cells was 8.9 for neutrophils, 16.4 for monocytes, and 6.7 for lymphocytes. These observations support the proposition that glutamine is required in much higher amounts than glucose to accomplish the total metabolic requirement of leukocytes. Under conditions closer to physiological when the availability of a variety of metabolites and vitamins is not restricted, glucose is the preferred substrate for lymphocytes, monocytes and neutrophils. PMID- 15338473 TI - Kupffer cell-derived prostaglandin E2 is involved in regulation of lipid synthesis in rat liver tissue. AB - Our recent studies suggest that Kupffer cells play a role in the physiological regulation of lipid metabolism of the adjacent hepatocytes. In the present study, we have tested the hypothesis that inhibition of Kupffer cells decreases prostaglandin E(2) (PGE(2)) release inside liver tissue, a phenomenon contributing to lipid accumulation in hepatocytes. PGE(2) secretion as well as lipid synthesis were assessed in precision-cut liver slices (PCLS) from rats previously treated with Kupffer cell inhibitors (GdCl(3) 10 mg kg(-1) body wt, i.v. injection and glycine 5% in diet). In addition, lipid synthesis was assessed in primary rat hepatocytes cultured in the absence or presence of PGE(2) (0.01, 1 and 10 microM). Inhibition of Kupffer cell activity by GdCl(3) decreases PGE(2) secretion by PCLS and resulted in a higher lipid synthesis. Since incubation with PGE(2) over 48 h decreases lipid synthesis from acetate in cultured hepatocytes, we propose that the lower PGE(2) secretion linked to Kupffer cell inhibition, partly explains a higher rate of synthesis of lipids with a subsequent accumulation in liver tissue, as previously shown in fasted rats. PMID- 15338474 TI - Influence of diadenosine tetraphosphate (Ap4A) on lipid metabolism. AB - Diadenosine polyphosphates (Ap(x)A) are physiologically released and may be partly involved in the pathogenesis of diabetes mellitus. Ap(4)A (diadenosine tetraphosphate) leads to an increase in blood glucose while it decreases insulin levels in plasma. A possible link between Ap(x)A and diabetes mellitus-associated diseases such as insulin resistance and hyperlipidemia (plasma free fatty acids, cholesterol and its biosynthesis, triacylglycerols) has not been investigated yet. Parameters such as free fatty acid and cholesterol content in blood were determined enzymically. The biosynthesis of cholesterol and triacylglycerols was determined in HepG2 cells using the radioactive precursor [(14)C]-acetate and by using gas chromatography. Plasma free fatty acids were significantly decreased 5 and 10 min after an Ap(4)A bolus (0.75 mg kg(-1) b.w.) given to rats. Plasma cholesterol was reduced 5 and 60 min after Ap(4)A administration. LPDS (lipoprotein-deficient serum)-stimulated cholesterol biosynthesis in HepG2 cells was significantly reduced after 1 h incubation with Ap(4)A. Triacylglycerol (TAG) biosynthesis in HepG2 cells was not significantly influenced by Ap(4)A; there was just a tendency for a concentration-dependent decrease in TAG levels. In conclusion Ap(4)A as a diabetogenetic compound is not likely to be responsible for the development of insulin resistance or of hyperlipidemia. Parameters such as free fatty acids, cholesterol and triacylglycerols are not elevated by Ap(4)A, but are even decreased. Ap(4)A seems to be involved in the development of diabetes mellitus by increasing blood glucose and decreasing plasma insulin as shown earlier, but not in diabetes mellitus-associated diseases such as insulin resistance or hyperlipidemia. PMID- 15338475 TI - Nitric oxide levels and antioxidant enzyme activities in jaundices of premature infants. AB - Free radicals are effective in the genesis of several diseases in the neonatal period. This study aimed to show the relationship between serum bilirubin levels and plasma nitric oxide and the activity of enzymes in the erythrocyte such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in premature infants. In the study, 20 premature infants with newborn jaundice were included and the control group was formed by 15 premature infants without jaundice. Venous blood samples were taken from all neonates in the study and control groups on the first day of hospitalization. Plasma nitric oxide levels and activities of SOD, GSH-Px and CAT enzymes in the erythrocytes were investigated in these samples. Plasma nitric oxide and serum bilirubin levels were found to be significantly higher in the study group (47.4 +/- 7.25 micromol l(-1), 18.41 +/- 3.28 mg dl(-1), respectively) than those in the control group (33.46 +/- 6.43 micromol l(-1), 4.35 +/- 0.60 mg dl(-1), respectively; p < 0.001). In addition, erythrocyte SOD, GSH-Px and CAT enzyme activities (724 +/- 78.61, 673 +/- 90.5, 63 +/- 12.8 U g(-1) Hb, respectively) were found to be significantly lower in the study group than those in the control group (1208 +/- 129.04, 1097.6 +/- 75.8, 99.06 +/- 12.4 U g(-1) Hb, respectively, p < 0.001). It was concluded that in the aetiology of hyperbilirubinemia, neonatal erythrocytes and nitric oxide reactions are affected differently and that erythrocyte haemolysis caused as a result of these effects may play a role in the aetiopathogenesis of unconjugated hyperbilirubinemia. Haemolysis may also be seen because of the inadequacy of the protection by erythrocytes against the cytotoxic effects of free radicals resulting from the lack of antioxidant enzymes in these cells. PMID- 15338477 TI - Pollen allergies in patients with systemic lupus erythematosus. PMID- 15338479 TI - Synovial density of frizzled 5-positive cells does not differ between patients with RA and OA and is independent of inflammation. PMID- 15338480 TI - Septicemia with Staphylococcus aureus, beta-hemolytic streptococci group B and G, and Escherichia coli in a patient with rheumatoid arthritis treated with a recombinant human interleukin 1 receptor antagonist (Anakinra). PMID- 15338481 TI - The no man's land of undifferentiated inflammatory polyarthritis. PMID- 15338482 TI - Window of opportunity. PMID- 15338483 TI - Issues in investigating the genetic epidemiology of disease expression. PMID- 15338484 TI - The quest for explanations for race/ethnic disparity in rates of use of total joint arthroplasty. PMID- 15338485 TI - Dating the "window of therapeutic opportunity" in early rheumatoid arthritis: accuracy of patient recall of arthritis symptom onset. AB - OBJECTIVE: A "window of therapeutic opportunity" has been hypothesized to be present in early rheumatoid arthritis (RA). To determine the date of this window, we must know the symptom-onset date of the RA. Patients participating in an observational study of early aggressive rheumatoid factor (RF) positive RA were evaluated to assess the accuracy of their recall of symptom-onset date by comparing the onset date they reported at the first visit with that reported on subsequent 6-monthly questionnaires. METHODS: One hundred eighty-six patients with early RA (at entry: median disease duration 5.8 mo, mean RF 413.8 +/- 630.7 IU/ml, 20.6 +/- 10.9 swollen and 23.7 +/- 13.4 tender joints) completed a self reported mailed questionnaire every 6 months for up to 5 years. As a part of each questionnaire, patients were asked to recall their RA symptom-onset date. These dates were then compared to the dates reported on the initial questionnaire. RESULTS: Thirteen months after symptom onset (i.e., about 6 mo after study entry) 61% of patients recalled the symptom-onset date (within 1 mo) that they had reported at baseline; the proportion decreased to 39% at 31 months and 25% after 70 months. During this period, the proportion overestimating RA duration remained about 20%, but the proportion underestimating it increased from 23% at 13 months to 39% at 31 months, and to 50% after 56 months. Patients with longer disease duration, less disease activity, and higher pain levels tended to be less accurate. CONCLUSION: Accuracy of recall of RA symptom-onset date by patients tends to decline over a period of 5 years. This should be taken into consideration when enrolling patients, when interpreting the findings of early RA clinical trials, and when attempting to ascertain the presence of a window of therapeutic opportunity. PMID- 15338486 TI - Effect of methotrexate therapy on bone mineral density and body composition in rat adjuvant arthritis. AB - OBJECTIVE: To test whether methotrexate (MTX) therapy of rat adjuvant arthritis (AA) prevents loss of bone mineral density (BMD) and loss of adipose and lean body mass compared to pair-fed controls with untreated rat AA (positive controls) and rats without AA (negative controls). METHODS: AA was induced by a Mycobacterium butyricum injection at the base of the tail of 5-week-old female Lewis rats. The MTX-treated group was injected with adjuvant and then treated twice weekly with MTX (1.0 mg/kg/wk intraperitoneally). To control for the effects of AA on appetite and weight, food given to control animals and MTX treated rats with AA was limited to that consumed by rats with untreated AA. At 42 days post-adjuvant injection, the animals were sacrificed and tibial BMD was measured. Body composition was analyzed for percentage fat, protein, ash, and water. RESULTS: There was no difference in ankle edema score or ankle width between the negative controls and MTX-treated group at necropsy. BMD was significantly higher in the negative controls versus positive controls and MTX treated and in MTX-treated versus positive controls. There was significantly less body fat and protein and greater body water in the positive controls and MTX group compared to the negative controls. CONCLUSION: MTX prevents loss of BMD in the tibia in the rat AA model compared to positive controls. While MTX is effective in lowering inflammation in rat AA, there are still significant losses in BMD and body composition, which may have implications for rheumatoid arthritis. PMID- 15338487 TI - Connective tissue metabolism in patients with unclassified polyarthritis and early rheumatoid arthritis. Relationship to disease activity, bone mineral density, and radiographic outcome. AB - OBJECTIVE: To assess the applicability of serum concentrations of markers of synovial inflammation, cartilage, and bone metabolism in relation to conventional markers of disease activity, bone mineral density (BMD) of the hand, and radiographic outcome. METHODS: Biochemical markers of collagen tissue metabolism were measured in 72 patients with symmetrically swollen and tender second and third metacarpophalangeal or proximal interphalangeal joints for at least 4 weeks and less than 2 years. At 2 years, 51 patients fulfilled the American College Rheumatology criteria for rheumatoid arthritis (RA) and 21 patients had unclassified polyarthritis. Patients with RA were divided into groups according to the mean disease activity and to magnetic resonance imaging and radiographically detected bone erosions in the hands. RESULTS: Patients with RA had significantly higher serum concentrations of matrix metalloproteinase-3 (MMP 3) at baseline and higher mean concentrations of serum MMP-3 and pyridinoline (Pyd) during the first 6 and 12 months than patients with unclassified polyarthritis. RA patients with persistent disease activity and erosive disease had significantly higher concentrations of serum MMP-3 and Pyd than patients with no disease activity or nonerosive disease. Significant mutual correlations between serum MMP-3 and Pyd and C-reactive protein and erythrocyte sedimentation rate were observed. The mean values of MMP-3 and Pyd correlated significantly to the alpha coefficient of the digital x-ray radiogrammetry (DXR-BMD). CONCLUSION: Serum MMP-3 and Pyd varied according to disease activity, periarticular osteoporosis measured by DXR, and radiographic outcome, and thus appear to supplement the conventional markers of disease activity for monitoring patients with RA. PMID- 15338488 TI - Indirect and total costs of early rheumatoid arthritis: a randomized comparison of combined step-down prednisolone, methotrexate, and sulfasalazine with sulfasalazine alone. AB - OBJECTIVE: To describe the effect of indirect costs for patients with early rheumatoid arthritis (RA) within the COBRA trial (Combinatietherapie Bij Reumatoide Artritis) on the cost-effectiveness of both therapies. Analyses of the efficacy and direct costs of the treatments have already been reported. METHODS: Patients with early RA selected for the 56-week trial were randomly assigned to prednisolone, methotrexate, and sulfasalazine (the COBRA combination) (n = 76, tapered after 28 weeks) or to sulfasalazine (SSZ; n = 79, of which 78 patients were evaluable) alone. The main efficacy outcomes were a pooled index and radiographic damage score in hands and feet, and utilities. Direct and indirect costs were measured (from a societal perspective) by means of cost diaries and interviews completed by patients during the intervention phase and the followup phase, each lasting 28 weeks. Differences in mean costs between groups and cost utility ratios were evaluated by applying nonparametric bootstrapping techniques. RESULTS: In the first 28 weeks, indirect costs per patient totaled US $2,578 and US $3,638 for COBRA and SSZ therapy, respectively (p = 0.09). The total costs were $5,931 and $7,853, respectively (p < 0.05). These differences were lost in the second 28 weeks. For the total period the mean total costs per patient were $10,262 and $12,788, respectively (p = 0.11). Sensitivity analyses showed robustness of the data. The point estimate of the cost per quality-adjusted life year based on the rating scale was negative at $-385, suggesting dominance of COBRA (more effect at lower cost). CONCLUSION: COBRA therapy adds additional disease control (improvements in disease activity, physical function, and rate of damage progression) at lower or equal cost compared to SSZ in early RA. PMID- 15338489 TI - Quality of life and economic burden of illness in very early arthritis. A population based study in southern Sweden. AB - OBJECTIVE: To measure health related quality of life (HRQOL) in patients with very early arthritis in a population based study in southern Sweden, and to compare HRQOL at baseline between the different diagnostic groups. Further, we investigated whether HRQOL at baseline correlated with the costs the patients incurred during the study. METHODS: Seventy-one adult patients with arthritis of less than 3 months' duration were referred from primary health care centers to rheumatologists. HRQOL was measured with the Arthritis Impact Measurement Scales (AIMS) and EuroQol at baseline. A comparison of HRQOL measures at baseline and the costs the patients incurred during the study was conducted in 56 of the patients. RESULTS: Twenty-seven (38%) patients had reactive arthritis (ReA), 17 (24%) undifferentiated arthritis, 15 (21%) rheumatoid arthritis (RA), 4 (6%) psoriatic arthritis, and the rest (11%) other diagnoses. Statistically significant differences were found between the 4 patient groups concerning the AIMS subscales of dexterity, household activity, activities of daily living (ADL) and pain, the patients with RA being most severely affected. There were no statistically significant differences between the 4 diagnosis groups concerning the EuroQol utility and EuroQol visual analog scale (VAS) scores. Of the AIMS subscales, the mobility, physical activity, household activity, ADL, and pain subscales correlated significantly with the incurred costs. Also the EuroQol utility scores and EuroQol VAS scores correlated significantly with the costs. Only the AIMS household activity subscale predicted the costs in the regression analysis. CONCLUSION: Patients with RA had significantly worse scores in the AIMS dexterity, household activities, ADL, and pain subscales compared to patients with other arthritides very early in the disease. The EuroQol generic quality of life instrument was less sensitive in detecting differences between patients with early arthritis than the disease-specific AIMS instrument. There was a correlation between the costs and the EuroQol utility scores and EuroQol VAS scores during the very first months of the disease, as well as with costs and the AIMS subscales of mobility, physical activity, household activity, ADL, and pain. PMID- 15338490 TI - Morning stiffness in patients with early rheumatoid arthritis is associated more strongly with functional disability than with joint swelling and erythrocyte sedimentation rate. AB - OBJECTIVE: To compare the level of morning stiffness in a cohort of patients with early rheumatoid arthritis (RA), assessed on a self-report questionnaire, to levels of patient self-report scores and clinical and laboratory variables. METHODS: A total of 337 patients with recent onset RA since 1998 were assessed for tender and swollen joint counts, erythrocyte sedimentation rate (ESR), physician global assessment, and radiographs of the hands and feet, as well as Multidimensional Health Assessment Questionnaire (MDHAQ) scores for functional disability, pain, fatigue, global status, morning stiffness, and number of symptoms. Regression models were used to estimate possible associations between these variables and morning stiffness. RESULTS: At study entry, 70 patients (21%) reported no morning stiffness, 52 (15%) reported morning stiffness < 15 minutes, 52 (15%) for 16-59 minutes, and 163 (49%) for >/= 1 one hour. At baseline and in longitudinal analyses, morning stiffness was significantly associated with functional disability scores on the MDHAQ and with other patient self-report data, and was associated at lower levels with swollen and tender joint counts and erythrocyte sedimentation rate (ESR). CONCLUSION: The degree of morning stiffness appears to reflect functional disability and pain more than traditional markers of inflammation such as joint counts and ESR in patients with early RA. Inclusion of morning stiffness as a marker of inflammatory activity in classification criteria for RA, inclusion criteria for most clinical trials in RA, and RA remission criteria, may be open to reassessment. PMID- 15338491 TI - Risk factors for methotrexate-induced abnormal laboratory monitoring results in patients with rheumatoid arthritis. AB - OBJECTIVE: To determine risk factors for methotrexate (MTX)-induced hepatic and hematologic laboratory abnormalities in patients with rheumatoid arthritis (RA). METHODS: Measurements of aspartate aminotransferase (AST), white blood cell counts, and platelet counts were collected in a database of patients with RA receiving MTX from 1991 through 2002. Potential risk factors for toxicity were recorded on each patient. RESULTS: Four hundred and eighty-one patients were followed for 2,323 person-years of MTX exposure. MTX was discontinued permanently because of abnormal laboratory test results in 22 patients (4.6%), the majority of whom (17/22, 77%) had elevated AST values. The body mass index (BMI) was significantly higher in those patients where MTX was permanently discontinued than in those in whom it was not (p < 0.03). Independent predictors of a significantly higher percentage of abnormal AST values were lack of folate supplementation (p < 0.001) and untreated hyperlipidemia (p < 0.02). Of the 17 patients in whom MTX was discontinued permanently because of an elevated AST value, 11/17 (65%) had either lack of folate supplementation or untreated hyperlipidemia. Hypoalbuminemia correlated independently with an increased percentage of abnormal platelet counts (p < 0.03). CONCLUSION: Lack of folate supplementation, untreated hyperlipidemia, and elevated BMI identified patients receiving MTX at risk for transaminase elevation, and low serum albumin was a risk factor for thrombocytopenia. Nonalcoholic fatty liver disease could be the underlying risk factor for transaminase elevation in patients with hyperlipidemia and obesity. PMID- 15338492 TI - Short term whole body retention in relation to rate of bone resorption and cartilage degradation after intravenous bisphosphonate (pamidronate) in rheumatoid arthritis. AB - OBJECTIVE: Bisphosphonates (BP) inhibit osteoclast-mediated bone resorption, and have been reported to decrease the rate of cartilage degradation. The anti resorptive effect of BP is determined by the amount of BP retained by the skeleton. In rheumatoid arthritis (RA) the uptake is not confined only to the skeleton, but BP is also retained in joints, which could have implications for dose regimens. We investigated the whole body retention (WBR) of pamidronate and its relationship to bone resorption and cartilage degradation in patients with active RA. METHODS: Twenty-six patients received placebo, 45 mg, or 90 mg intravenous pamidronate. Serum and urine samples were collected before and for 12 days after drug administration. Rate of bone resorption was assessed by the biochemical markers: serum carboxy terminal cross-linked telopeptide of type I collagen, urinary carboxy terminal cross-linked telopeptide of type I collagen normalized to creatinine and urinary amino-terminal telopeptide of type I collagen normalized to creatinine; and rate of cartilage degradation by urinary carboxy terminal telopeptide of type II collagen normalized to creatinine. WBR was derived from urinary excretion of pamidronate data. RESULTS: Pamidronate induced a rapid and sustained decrease in the level of biochemical markers of bone resorption and cartilage degradation. The mean WBR of pamidronate was 69% of the administered dose, and showed a remarkably wide range (41-96%). The decrease in rate of bone resorption, but also rate of cartilage degradation appeared to be related to the WBR of pamidronate. CONCLUSION: This is the first study in which the effect of BP treatment has been studied in relation to the amount of BP retained by the body in patients with active RA. The total amount of BP retained by the body shows a remarkably wide range and is comparable with literature on patients with osteoporosis. The apparent relationships between the amount of BP retained by the body and the effect could have implications for therapeutic regimens in patients with RA. PMID- 15338493 TI - Partner participation in cognitive-behavioral self-management group treatment for patients with rheumatoid arthritis. AB - OBJECTIVE: To determine if participation of the spouse of patients with rheumatoid arthritis in cognitive-behavioral oriented self-management training aimed at improving disease related cognitions and coping with pain has additional benefits for the patients. METHODS: A total of 59 couples were randomly assigned to 2 conditions. In the experimental condition the couples participated in a group program aimed at restructuring disease related cognitions and decreasing passive coping. In the control condition, only the patient participated. Disease status, self-reported physical and psychological functioning, coping, disease related cognitions, and marriage characteristics were assessed prior to the intervention and 2 weeks and 6 months postintervention. A general linear model with repeated measures was used to test for differences between conditions. RESULTS: In both conditions, similar positive changes in disease activity, cognitions, coping, and physical and psychological functioning were observed. Patients reported a decrease in potential support. There were no differences between conditions. However, at the followup assessment patients in the experimental condition reported more improvement of disease related communication with their spouse. CONCLUSION: No evidence was found for additional beneficial effects of spouse participation in the cognitive-behavioral oriented self management group treatment. PMID- 15338494 TI - Dyslipoproteinemia in patients with active rheumatoid arthritis: effects of disease activity, sex, and menopausal status on lipid profiles. AB - OBJECTIVE: To investigate the lipid profiles in patients with active rheumatoid arthritis (RA) and to assess the relationship of inflammatory disease activity markers, sex, and menopausal status with lipid profiles. METHODS: Three groups of patients with active RA (n = 184) were studied: men (n = 61, mean age 50.8 +/- 4.81 yrs), premenopausal women (n = 58, mean age 39.2 +/- 2.44 yrs), and postmenopausal women (n = 65, mean age 60.4 +/- 2.14 yrs), and healthy controls (n = 161): men (n = 65, mean age 50.9 +/- 3.42 yrs), premenopausal women (n = 47, mean age 40.3 +/- 1.66 yrs), and postmenopausal women (n = 49, mean age 61.3 +/- 3.16 yrs). We measured fasting plasma levels of total cholesterol (TC), triglyceride (TG), HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), lipoprotein (a) [LP(a)], apolipoprotein A1 (apo A1), apolipoprotein B (apo B), and erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). RESULTS: Male RA patients had significantly higher apo B/apo A1 and LP(a) and lower HDL-C than male controls. Female RA patients had significantly higher TC, LDL-C, and LP(a) than female controls. Premenopausal RA patients had significantly higher LDL-C, TC/HDL-C, LDL-C/HDL-C, and apo B/apo A1 and lower TG and HDL-C than premenopausal controls. Postmenopausal RA women had significantly higher TG and LP(a) and lower TC than postmenopausal controls. Female RA patients had higher HDL-C, apo A1, and TC/HDL-C and lower apo B/apo A1 than male RA patients. Postmenopausal RA patients had significantly higher TC, TG, TC/HDL-C, apo B, LP(a), and LDL-C/HDL-C than premenopausal RA patients. CRP correlated positively with TC/HDL-C, LDL-C/HDL-C, and apo B/apo A1 and negatively with HDL-C in male RA patients. In female RA patients CRP had positive correlation with TC/HDL-C and LDL-C/HDL-C and negative correlation with HDL-C. CONCLUSION: These findings suggest that patients with active RA have altered lipid profiles and that disease activity, sex, and menopausal status affect lipid profiles, and these would be expected to change the pattern of atherosclerotic events in RA. PMID- 15338495 TI - Utility of red amsler grid screening in a rheumatology clinic. AB - OBJECTIVE: To ascertain the false positive rate of red Amsler grid screening in an outpatient rheumatologic setting. METHODS: Red Amsler grid screening was performed on 170 consecutive patients presenting with rheumatoid arthritis or systemic lupus erythematosus. Patients with abnormal screening results were referred for ophthalmologic evaluation and formal visual field testing. RESULTS: Nineteen patients identified scotomata on red Amsler grid. Despite numerous recall attempts, only 9 of the 19 patients with scotomata presented for ophthalmologic evaluation. The positive predictive value was 5-58%. Roughly 6-11% of all patients screened had a false positive screening test. CONCLUSION: While red Amsler grid screening may allow rheumatologists to identify patients who are more likely to have hydroxychloroquine ocular toxicity, an abnormal red Amsler grid screening is in itself an unreliable predictor of toxicity. PMID- 15338496 TI - Angiotensin-converting enzyme (insertion/deletion) and endothelial nitric oxide synthase polymorphisms in patients with systemic lupus erythematosus. AB - OBJECTIVE: Genetic polymorphisms in the angiotensin-converting enzyme (ACE) and endothelial nitric oxide synthase (eNOS) are linked with expression and/or progression of renal disease. We hypothesized that polymorphisms in the genes coding for ACE and eNOS may influence the development and/or progression of systemic lupus erythematosus (SLE) and lupus nephritis given their linkage with other renal diseases. METHODS: DNA from patients with SLE (n = 227) and their age and sex matched controls (n = 275) from the Carolina Lupus (CLU) Study cohort was assessed for ACE and eNOS polymorphisms. Seventy patients had biopsy-proven lupus nephritis. Two different eNOS polymorphisms (eNOS promoter T-786(R)C nucleotide substitution and eNOS 27 base pair tandem repeat in intron 4) and the ACE insertion/deletion (I/D) polymorphism in intron 16 were examined by restriction fragment length polymorphism-polymerase chain reaction. RESULTS: Allele frequency of the eNOS polymorphisms varied significantly between SLE patients and controls. There was no association of these polymorphisms with lupus within ethnic groups. We found no association of the polymorphism with the development of renal disease. No association was observed for the ACE I/D polymorphism with SLE or nephritis, or with ethnicity or sex. CONCLUSION: eNOS genetic polymorphisms differed significantly across ethnic groups. There was no significant increased risk of SLE and/or lupus nephritis associated with eNOS or ACE polymorphisms in either the African American or Caucasian groups compared to ethnically matched controls. These studies emphasize the need to control for ethnicity when investigating genetic polymorphisms and disease. PMID- 15338497 TI - Increased incidence of cervical intraepithelial neoplasia in women with systemic lupus erythematosus treated with intravenous cyclophosphamide. AB - OBJECTIVE: To determine if the incidence of cervical intraepithelial neoplasia (CIN) is increased in immunosuppressed women with systemic lupus erythematosus (SLE). METHODS: Women with SLE were consecutively recruited from University of Michigan outpatient rheumatology clinics. Women with abnormal cervical smears at screening were excluded. Cervical smears were obtained at baseline and at 3 and 7 years. Cervical biopsies confirmed cytologic abnormalities (CIN I-III), and were scored by pathologists in blinded fashion. Data were analyzed according to treatment group: (1) prednisone; (2) azathioprine (AZA); (3) intravenous cyclophosphamide (IVCYC); and (4) IVCYC + AZA + prednisone. RESULTS: Sixty-one of 89 women screened were eligible for enrollment. The overall 3-year incidence of CIN was 9.8%. Stratified by treatment group, the 3-year incidence of CIN was 0/23 (0%) in prednisone treated patients, 0/4 (0%) in AZA treated patients, 2/8 (25%) in IVCYC treated patients, and 4/26 (15%) in CYC + AZA + prednisone treated patients. A dose relationship was observed between cumulative IVCYC exposure and CIN; each increase of 1 g of IVCYC exposure corresponded to a 13% increased risk of CIN (p = 0.04). At 7 years, 45 patients remained under followup and 6 patients had died of unrelated causes. No cases of CIN were observed at 7 years, although there were 2 cases of atypical squamous cells of unknown significance and one case of condyloma. CONCLUSION: IVCYC + prednisone therapy for SLE is significantly associated with development of CIN. PMID- 15338498 TI - Immunogenetic features in 120 Japanese patients with idiopathic inflammatory myopathy. AB - OBJECTIVE: To examine the role of HLA-DRB1 and tumor necrosis factor (TNF) promoter genotypes in the development and the autoantibody profiles of idiopathic inflammatory myopathy (IIM) in Japanese patients. METHODS: HLA-DRB1 and TNF promoter genotypes were determined, and serum antinuclear autoantibodies were identified in 120 adult Japanese patients with IIM [72 with dermatomyositis (DM), 30 with polymyositis (PM), 18 with myositis overlapping with other collagen vascular diseases], as well as in 265 controls. RESULTS: Forty-two patients (35%) were positive for myositis-specific autoantibodies (MSA), including 37 (31%) for anti-aminoacyl-tRNA synthetase (ARS) autoantibodies. Allele carrier frequency of HLA-DRB1*0803 was increased in the patients with IIM [p = 0.02, corrected p (pc) NS, 23% vs 14%, odds ratio (OR) = 1.9 (95% confidence interval, CI = 1.1-3.2)], with PM [p = 0.006, pc NS, 33%, OR 3.1 (95% CI 1.3-7.1)], and with anti-ARS autoantibodies [27%, p = 0.04, OR 2.3 (95% CI 1.0-5.1)] compared with controls. DRB1*0405 was increased in patients with anti-ARS autoantibodies compared with controls [41% vs 25%, p = 0.04, pc NS, OR 2.1 (95% CI 1.0-4.3)]. TNF promoter genotype was associated with the presence of interstitial lung disease (ILD). The carriage of a TNF-a haplotype formed by -1031C, -863A, and -857C was increased in the patients with ILD versus those without ILD [33% vs 18%, p = 0.05, pc NS, OR 2.3 (95% CI 0.94-5.5)]. CONCLUSION: HLA-DRB1 alleles were associated with development of IIM and MSA in a Japanese population. PMID- 15338499 TI - Concordance of disease severity among family members with ankylosing spondylitis? AB - OBJECTIVE: The heritability of disease activity and function in ankylosing spondylitis (AS) have been estimated at 0.51 and 0.63 (i.e., 51% and 63%), respectively. We examined the concordance of disease severity among family members in terms of disease activity, function, radiological change, prevalence of iritis, and juvenile onset. METHODS: Disease activity and functional impairment due to AS were studied using the Bath AS Disease Activity Index (BASDAI) and Functional Index (BASFI) self-administered questionnaires; radiographic involvement was measured using the Bath AS Radiology Index (BASRI) scale. Familial correlation of BASDAI and BASFI was assessed in 406 families with 2 or more cases, using the program PAP. Parent-child and sibling-sibling concordance for iritis and juvenile AS were also studied in these families. Heritability of radiological disease severity based on the BASRI was assessed in 29 families containing 60 affected individuals using the program SOLAR. RESULTS: Correlations between parent-child pairs for disease activity and function were 0.07 for both. Correlations between sibling pairs for disease activity and function were 0.27 and 0.36, respectively. The children of AS parents with iritis were more likely to develop iritis [27/71 (38%)] than children of non-iritis AS parents [13/70 (19%)] (p = 0.01). Parents with JAS were more likely to have children with JAS [17/30 (57%) compared to non-JAS parents 34/111 (30%)] (p = 0.002). The heritability of radiological disease severity based on the BASRI was 0.62. CONCLUSION: While correlation in severity between parent and child is poor, siblings do resemble each other in terms of severity, supporting the findings of segregation studies indicating significant genetic dominance in the heritable component of disease activity. Significant parent-child concordance for iritis and juvenile disease onset suggest that there are genetic risk factors for these traits independent of those determining the risk of AS itself. The finding of significant heritability of radiological change (BASRI) provides support using an objective measure for the observed heritability of the questionnaire-assessed disease severity scores, BASDAI and BASFI. PMID- 15338500 TI - Measurement of spinal mobility in ankylosing spondylitis: comparison of occiput to-wall and tragus-to-wall distance. AB - OBJECTIVE: To investigate if the tragus-to-wall distance (TWD) is more reliable compared to the occiput-to-wall distance (OWD) as a measurement for thoracic spine extension in patients with ankylosing spondylitis (AS). METHODS: Data from the OASIS cohort, an international longitudinal observational study on outcome in AS, were used. Measurements of OWD and TWD were performed at baseline and at 6, 12, 18, and 24 months. Paired data of Tx and Tx+6 months were used to perform test-retest measurements (intraclass correlations, limits of agreement, and interperiod correlation matrix). Bland and Altman plots were constructed to investigate the agreement between both observations, assuming that there was no true change between 0 and 6 months. To investigate whether a change in disease activity would have influenced the results, limits of agreement were calculated in a subgroup of patients with a stable Bath Ankylosing Spondylitis Disease Activity Index (BASDAI; defined as a maximum BASDAI change of +/- 1) between T0 and T6 and compared with the results of the whole group. Limits of agreement were also calculated for kyphosed patients only. RESULTS: The test-retest intraclass correlations were between 0.94 and 0.96 for OWD and between 0.93 and 0.95 for TWD. The direct measurement-remeasurement correlation calculated by extrapolation of the interperiod correlation regression line was 0.92 for OWD and 0.90 for TWD. OWD and TWD showed comparable reliability on the entire value of scores. The lower 95% limit of agreement was between -3.4 cm and -2.5 cm for OWD and between 3.4 cm and -3.1 cm for TWD. The upper limit of agreement was between 3.1 cm and 4.2 cm for OWD and between 2.9 cm and 3.9 cm for TWD. In all patients as well as in kyphosed patients only, limits of agreement were comparable between OWD and TWD. The patterns of the scatterplots according to Bland and Altman were similar for OWD and TWD. Measurement error was more pronounced in kyphosed patients compared to patients with a normal thoracic extension. However, over the entire range of kyphosis, measurement error was similar. CONCLUSION: OWD and TWD are equally reliable in assessing thoracic spine extension. Although the TWD is in general easier to perform in AS patients compared to OWD, we recommend the OWD measurement over TWD: in OWD measurement a value of zero easily distinguishes patients with normal thoracic spine extension from kyphosed patients. PMID- 15338501 TI - Vascular endothelial growth factor gene polymorphisms in Behcet's disease. AB - OBJECTIVE: To evaluate potential associations of vascular endothelial growth factor (VEGF) gene polymorphisms with Behcet's disease (BD) and disease expression. METHODS: Case patients were 122 consecutive Italian patients with BD followed at the Rheumatology, Ophthalmology, and Neurology Units in Bologna, Ferrara, Milano, Palermo, Potenza, Prato, Reggio Emilia, and Trento over a 3-year period (1997-99) and who satisfied the International Study Group criteria for BD. Also selected as a control group were 200 healthy age and sex matched blood donors. All patients with BD and controls were genotyped by polymerase chain reaction and allele-specific oligonucleotide techniques for +936 C/T (rs3025039) and -634 C/G (rs2010963) mutations and for an 18 base pair (bp) insertion/deletion (I/D) polymorphism at -2549 of the the VEGF promoter region. In vitro release of VEGF by peripheral blood mononuclear cells (PBMC) was investigated by ELISA in healthy controls homozygous for the polymorphisms studied. RESULTS: The carriage rates of the alleles I and -634C were significantly more frequent in patients with BD than in healthy controls [p corr = 0.036, OR 1.8 (95% CI 1.1-2.9) and p corr = 0.05, OR 1.8 (95% CI 1.1-3.0), respectively]. While the distribution of allele +936T was similar in patients with BD and healthy controls, its frequency was significantly higher in BD patients with posterior uveitis/retinal vasculitis than in those without (p = 0.022, OR 2.4, 95% CI 1.1-5.0). Lipopolysaccharide-stimulated VEGF production from PBMC of healthy subjects was higher in II homozygous than in DD homozygous. CONCLUSION: Our data indicate that carriers of -634C and I alleles are associated with susceptibility to developing BD. PMID- 15338502 TI - Surgical approach for total hip arthroplasty: direct lateral or posterior? AB - OBJECTIVE: To determine the risks of prosthesis dislocation, postoperative Trendelenburg gait, and sciatic nerve palsy after a posterior approach compared to a direct lateral approach for adult patients undergoing total hip arthroplasty (THA) for primary osteoarthritis (OA). METHODS: Medline, Embase, CINHAL, and Cochrane databases were searched until August 2003. All published trials comparing posterior and direct lateral surgical approaches to THA in adults with a diagnosis of primary hip osteoarthritis were collected. Retrieved articles were assessed independently for their methodological quality. RESULTS: Four prospective cohort studies involving 241 participants met the inclusion criteria. Regarding dislocation rate, no significant difference between posterior and direct lateral surgical approach was found (relative risk 0.35). The presence of postoperative Trendelenburg gait was not significantly different between surgical approaches. The risk of nerve palsy or injury was significantly higher with the direct lateral approach (relative risk 0.16). However, there were no significant differences when comparing this risk nerve by nerve, in particular for the sciatic nerve. Of the other outcomes considered, only the average range of internal rotation in extension of the hip was significantly higher (weighted mean difference 16 degrees ) in the posterior approach group (mean 35 degrees, SD 13 degrees ) compared to the direct lateral approach (mean 19 degrees, SD 13 degrees ). CONCLUSION: The quality and quantity of information extracted from the trials performed to date are insufficient to make a firm conclusion on the optimum choice of surgical approach for adult patients undergoing primary THA for OA. PMID- 15338503 TI - Cost-effectiveness of treatment strategies for osteoarthritis of the knee in Taiwan. AB - OBJECTIVE: To evaluate the cost-effectiveness of 3 treatment strategies for osteoarthritis (OA) of the knee: naproxen, celecoxib, and hyaluronan. METHODS: We developed a decision model to estimate the costs and effectiveness of 3 treatment strategies: 250 mg naproxen 3 times daily for 26 weeks, 100 mg celecoxib twice daily for 26 weeks, and 25 mg hyaluronan by intraarticular injection once per week for 5 weeks followed by conventional treatment for 21 weeks. The probabilities and utility data were obtained by surveying the literature and consulting experts. Cost data were obtained from insurance reimbursement data of National Taiwan University Hospital and were converted to 2002 US dollars. The timeframe of the decision tree was 26 weeks. Outcomes were expressed in aggregated costs, quality-adjusted life-years (QALY), and the incremental cost effectiveness ratio (ICER). Sensitivity analyses were performed on most variables. RESULTS: The expected total costs for the naproxen, celecoxib, and hyaluronan strategies were US$498.98, US$547.80, and US$678.00, respectively. The ICER of the celecoxib strategy compared with the naproxen strategy was US$21,226 per QALY gained. The ICER of the hyaluronan strategy versus the celecoxib strategy was US$42,000 per QALY gained. The ICER of the hyaluronan strategy decreased to about US$25,000 per QALY gained if the weekly treatment cost of hyaluronan was decreased to US$31. CONCLUSION: Celecoxib treatment results in a reasonable cost-effectiveness ratio for patients with OA of the knee. Hyaluronan treatment, however, may not be an economically attractive choice under the current healthcare scenario in Taiwan. PMID- 15338504 TI - Gastroduodenal safety and tolerability of lumiracoxib compared with Ibuprofen and celecoxib in patients with osteoarthritis. AB - OBJECTIVE: To compare the incidence of gastroduodenal ulcers in patients with osteoarthritis (OA) treated with therapeutic doses of the novel COX-2 selective inhibitor, lumiracoxib (COX189, Prexige), and the standard nonsteroidal antiinflammatory drug (NSAID) ibuprofen. The COX-2 selective inhibitor celecoxib was included as an active control. METHODS: In this randomized, multicenter, double-blind, parallel-group study, eligible patients were randomized to receive lumiracoxib 200 mg (n = 264) or 400 mg (n = 260) once daily (qd), ibuprofen 800 mg (n = 260) 3 times daily (tid), or celecoxib 200 mg qd (n = 258) for 13 weeks. The incidence of gastroduodenal ulcers and erosions was determined by endoscopy prior to randomization, and after 4 weeks and 13 weeks of treatment (end of study). Frequencies of adverse events were also recorded. RESULTS: The cumulative incidence of gastroduodenal ulcers >/= 3 mm in diameter was significantly lower in the lumiracoxib groups (200 mg: 4.3%; 400 mg: 4.0%) than in the ibuprofen group (15.7%; p < 0.001) and similar to the celecoxib group (3.2%). In the ibuprofen group, a significantly greater number of patients (6.0%) had > 10 gastroduodenal erosions compared with lumiracoxib 200 mg (1.2%; p < 0.01), lumiracoxib 400 mg (1.6%; p < 0.05), and celecoxib (2.4%; p < 0.05). A greater number of patients in the ibuprofen group discontinued treatment due to an adverse event compared with both lumiracoxib groups and the celecoxib group. CONCLUSION: In patients with OA, lumiracoxib 200 mg or 400 mg qd was associated with a significantly lower risk of gastroduodenal ulceration than ibuprofen 800 mg tid, and was similar to celecoxib 200 mg qd. PMID- 15338505 TI - Ethnic differences in health preferences: analysis using willingness-to-pay. AB - OBJECTIVE: Racial and ethnic differences in health services utilization are well recognized, but the explicit contribution of access to care, physician bias, and patient preferences to these disparities remains unclear. We investigated whether preferences for improvements in health vary among ethnic groups. We chose to assess preferences for osteoarthritis (OA) of the knee because significant differences have been observed in the utilization of total knee arthroplasty among ethnic groups, and because it is an elective procedure, where individual preferences have a major role in decision-making. METHODS: A survey using willingness-to-pay (WTP) methodology was conducted to elicit preferences for improvement in severe and mild OA and for 5 non-health items; data were collected from 193 white, African American, and Hispanic individuals over the age of 20 years. Multivariate regression analyses were used to determine whether WTP varied across racial/ethnic groups. RESULTS: WTP as a percentage of income for each of the 3 scenarios was highest for whites, intermediate for Hispanics, and lowest for African Americans (e.g., 32.9%, 26.4%, and 16.7% for mild OA). Controlling for income, differences in log WTP between African Americans and whites were significant in multivariate regression analyses, whereas values for Hispanics and whites did not differ significantly. Race/ethnic group variables explained a relatively large (21-30%) part of the variation in log WTP. CONCLUSION: The findings suggest that ethnic differences in health valuation and preferences contribute to the observed disparities in health services utilization of elective procedures such as total knee arthroplasty. PMID- 15338506 TI - Assessment of disease severity (in terms of function) using the internet. AB - OBJECTIVE: To determine whether visual analog scales (VAS) can be used over the Internet to assess the patient with ankylosing spondylitis (AS) accurately or if the use of this different medium will affect the results. METHODS: Patients with AS (n = 50) attending a physiotherapy/educational course completed both an Internet based and a paper based version of the Bath Ankylosing Spondylitis Functional Index (BASFI) that uses VAS. The Internet version was completed twice to assess intrarespondent variation reliability and compared with the paper version to assess interrespondent variation reliability. Patients were also asked to assess ease of use and to suggest changes to the Internet version. RESULTS: The interclass coefficient of intra- and interrespondent reliability were 0.989 (p < 0.001) and 0.976 (p < 0.001), respectively. There was a 3% difference in assessments carried out over the Internet compared to those on paper and a 2% difference in repeatability of Internet assessed questionnaires. Bland and Altman plots showed a mean difference between paper compared to the Internet version was 0.0156 with 95% limits of agreement at -1.07 to 1.03. More than half the participants reported that the Internet version was easier to complete than the paper version (96% rated usability as 1 on a scale of 1-10, with 1 being extremely easy and 10 being impossible). CONCLUSION: Assessment of disease severity by VAS may be accurately carried out over the Internet. This means that the evaluation of disease status and the longterm followup of people in different countries and perhaps in different languages may now be possible, using the Internet. PMID- 15338507 TI - Deaths from arthritis and other rheumatic conditions, United States, 1979-1998. AB - OBJECTIVE: To analyze US trends in deaths from arthritis and other rheumatic conditions (AORC). METHODS: Multiple cause of death tapes from the National Center for Health Statistics from 1979 to 1998 were reviewed. Age, sex, and race specific death rates were calculated. RESULTS: During 1979-1998, the annual number of AORC deaths rose from 5537 to 9367. In 1979, the crude death rate from AORC was 2.46 per 100,000 population; by 1998, it was 3.48. Rates age standardized to the year 2000 population were 2.75 and 3.51, respectively. Annual crude and age-standardized death rates were higher among women than men and higher among blacks than whites and increased for all groups over the 20 years. Death rates were dramatically higher with increasing age. Three categories of AORC accounted for almost 80% of deaths: diffuse connective tissue diseases (34%), other specified rheumatic conditions (23%), and rheumatoid arthritis (22%). CONCLUSION: There are marked age, sex, and race-specific disparities in AORC death rates. AORC death rates may be underestimated because of (1) nonrecognition of inflammatory arthritis and (2) attribution of cause of death to conditions made more likely by arthritis, e.g., cardiovascular disease, or to complications from arthritis therapy. Further research into the causes of the disparities in death rates and the increase in death rates for men, women, blacks, and whites is necessary. PMID- 15338508 TI - Determination of anti-cyclic citrullinated peptide antibodies in the sera of patients with juvenile idiopathic arthritis. AB - OBJECTIVE: Anti-cyclic citrullinated peptide (anti-CCP) antibodies have been found in sera of 76% of patients with rheumatoid arthritis (RA), mainly in rheumatoid factor (RF) positive patients, with a specificity of 96%. We evaluated the presence of anti-CCP antibodies in patients with juvenile idiopathic arthritis (JIA) and assessed the possibility of synthetic citrullinated peptides as antigenic determinants in JIA. METHODS: The presence of anti-CCP antibodies was determined using 3 synthetic citrullinated peptide variants and 2 commercial kits (Inova Diagnostics and Axis-Shield Diagnostics) optimized for detecting JIA specific antibodies in serum by an ELISA based assay. We evaluated 66 patients with JIA (16 RF positive polyarthritis, 18 RF negative polyarthritis, 19 oligoarthritis, and 13 systemic arthritis). We also tested 9 adult RA patients, 34 patients with systemic lupus erythematosus (SLE), and 25 healthy persons as controls. RESULTS: Significant concentrations of anti-CCP antibodies were detected in the majority of RF positive JIA patients with polyarthritis. Using the 2 synthetic linear peptides, 12/16 (75%) were positive; 9/12 (75%) were positive with the Inova kit and 9/10 (90%) were positive with the Axis-Shield kit. However, utilizing the synthetic linear peptides, significant concentrations of anti-CCP antibodies were detected in 51/66 (77%) JIA patients, including 15/18 (83%) RF negative polyarthritis, 16/19 (84%) oligoarthritis, and 8/13 (62%) systemic arthritis patients. No healthy control showed elevated antibody levels. In contrast, 4/9 (44%) patients with adult RA and 2/6 (33%) with SLE had elevated anti-CCP levels. The synthetic cyclic variant cfc-1-cyc yielded significant anti CCP levels for 13/14 (93%) patients with RF negative polyarthritis, 6/10 (60%) with oligoarthritis, and 3/7 (43%) with systemic arthritis, and 8/9 (88%) RF positive patients. No healthy control had increased anti-CCP levels. However, 4/9 (44%) adult RA and 9/34 (26%) SLE patients were found to have elevated anti-CCP levels. Using the Inova and Axis-Shield kits, much smaller percentages were found in the RF negative patients, with only 4/16 (25%) in the oligoarthritis and RF negative polyarthritis patients with the Inova kits and 0/25 (0%) by the Axis Shield kits. The Inova kit revealed elevated anti-CCP antibodies in 5/9 (56%) adult RA patients and in 8/34 (24%) SLE patients. No healthy control had elevated anti-CCP antibodies. However, the Axis-Shield kits did not detect anti-CCP antibodies in adult RA (0/9) or SLE (0/34) patients. Moreover, 0/25 (0%) healthy individuals exhibited anti-CCP levels. The presence of anti-CCP antibodies correlated more frequently with the presence of RF. CONCLUSION: This study confirms the presence of anti-CCP antibodies in patients with JIA, especially those with RF positive polyarthritis, by all ELISA based methods. Use of synthetic peptides also revealed anti-CCP antibodies in a percentage of RF negative patients with polyarthritis, oligoarthritis, and systemic arthritis; there was a loss in specificity, but an increase in sensitivity. These results suggest that antibodies to these antigenic peptides may be markers for JIA, and indicate a possible role of citrulline-containing epitopes in the pathogenesis of JIA. PMID- 15338509 TI - Cardiopulmonary exercise testing in juvenile idiopathic arthritis. AB - OBJECTIVE: To assess aerobic fitness and exercise capacity in patients with juvenile idiopathic arthritis (JIA) and to determine subgroup differences. METHODS: Thirty-four patients diagnosed with JIA and 21 healthy sedentary volunteers were studied. Aerobic fitness was determined by measuring peak power and peak oxygen uptake (VO2peak) during an incremental cycling test. The patient group consisted of systemic JIA (n = 8), polyarticular JIA (n = 13), oligoarticular JIA (n = 7), and enthesitis-related arthritis (ERA, n = 6). Results from different subgroups of JIA were compared to determine subgroup differences. RESULTS: All subjects tolerated maximal exercise testing well. The JIA group had lower aerobic fitness than controls. In our comparison of JIA subgroups, we found no significant differences in cardiopulmonary measures. The ERA group had higher aerobic capacity than other subgroups. There was no difference in exercise capacity between patients with active disease (n = 10) and those in remission (n = 24). CONCLUSION: We suggest that heterogeneity in VO2peak levels among JIA patients is due to subgroup differences. Exercise programs for improvement of aerobic fitness should be individualized or at least be modified according to different subgroups. PMID- 15338510 TI - Juvenile idiopathic arthritis: parent-child discrepancy on reports of pain and disability. AB - OBJECTIVE: To examine the incidence and nature of disagreements about pain and functional disability between parents and their children with juvenile idiopathic arthritis (JIA) and to identify demographic and psychosocial predictors of parent child disagreement about pain and functional disability. METHODS: Participants comprised 63 children 8-16 years of age (mean 12.36 +/- 2.61) and their parents, followed as part of a longitudinal study of pain in children. During routine rheumatology clinic visits, children and their parents completed validated measures of pain, depressive symptoms, and functional disability. RESULTS: Parents and children often disagreed as to the frequency and intensity of pain and to the degree of disability caused by arthritis. Child depressive symptoms (p < 0.01) and parental perceptions of child limitations (p < 0.02) predicted parent child disagreement about the frequency of the child's pain. Parental perceptions of child limitations also predicted parent-child disagreement about the child's level of functional disability (p < 0.04). Those children who estimated their level of disability to be different than their parents' rating also were more depressed compared to children who agreed with their parents about their level of disability (p < 0.01). CONCLUSION: Discrepancy between parent and child reports of pain and disability in children with JIA is common. Findings suggest that such disagreements in reporting of pain and functional disability by parents and their children with JIA are associated with underlying depressive symptoms in children. PMID- 15338511 TI - HLA-DRB1*15021 is the predominant allele in Japanese patients with juvenile dermatomyositis. AB - OBJECTIVE: To investigate HLA molecules and genes in Japanese patients with juvenile dermatomyositis (JDM). METHODS: Twelve patients (8 girls and 4 boys) with ages of onset between 3 and 15 years were included. HLA class I antigen phenotypes were serologically typed by the Terasaki-NIH standard method. DNA was extracted from peripheral blood leukocytes using the phenol-chloroform extraction procedure, and stored at -70 degrees C until use. Genomic DNA for HLA-DRB1, HLA DQA1, and HLA-DQB1 alleles in JDM patients and controls was determined by the direct sequence method. RESULTS: HLA-A24 and B52 were each detected in 7 cases (OR = 0.86, 5.02, p = 0.930, 0.006, respectively). HLA-DRB1*15021 was observed in 7 patients. This was significantly more frequent than occurred in the controls (OR = 5.72, p = 0.002). Seven patients out of 12 (58%) had the combination HLA B52, DRB1*15021, DQA1*0103, and DQB1*0601. CONCLUSION: Our results suggest that the susceptibility gene for JDM either is HLA-DRB1*15021 or is present near the HLA-DRB1 locus. This differs from previous reports that describe the association with HLA-DQA1*0501 in Caucasian patients with JDM. The combination HLA-B52, DRB1*15021, DQA1*0103, and DQB1*0601 may contribute to the pathogenesis of JDM in Japanese patients. PMID- 15338512 TI - Systemic lupus erythematosus and calciphylaxis. AB - Calciphylaxis, a rare condition seen in association with endstage renal disease, is characterized by the appearance of painful, indurated plaques, ecchymosis, ulceration, and eschar formation. We describe a patient with systemic lupus erythematosus, endstage renal disease, and skin lesions first diagnosed as lupus profundus with vasculitis. Further investigation confirmed the diagnosis of extensive calciphylaxis. PMID- 15338513 TI - Paraneoplastic polyarthritis in an ovarian teratoma. AB - A 34-year-old woman presented to the rheumatology clinic with severe low back pain and arthralgia; later she developed bilateral knee pain and swelling, with limitation in ambulation and minimal improvement with nonsteroidal antiinflammatory drugs. Two weeks later she developed pain on the volar aspect of the right wrist and on the hypothenar region of the right hand. Examination showed swelling and tenderness of the right hypothenar region, tenderness and decreased flexion and extension of the right wrist, and bilateral knee effusions. The combination of arthritis and tenosynovitis raised the possibility of an ovarian tumor. A pelvic ultrasound revealed a complex, hyperechoic ovarian mass consistent with a cystic teratoma. Four weeks after removal of the teratoma, the polyarthritis and related symptoms resolved without therapy. PMID- 15338514 TI - Multiple spontaneous visceral hematomas revealing polyarteritis nodosa. AB - Polyarteritis nodosa is a rare life-threatening disease characterized by necrotizing vasculitis of small and median arteries. We describe the exceptional case of a 28-year-old man with successive spontaneous visceral hematomas of the kidney, bladder, and liver. Arteriography was performed for a recent spontaneous hepatic hematoma and a microaneurysm was detected, allowing the diagnosis of polyarteritis nodosa and prescription of appropriate treatment. PMID- 15338515 TI - Disseminated Prototheca wickerhamii infection with arthritis and tenosynovitis. AB - Achloric algae of the Prototheca species are a rare cause of infection in humans. These infections are usually localized to the skin, olecranon bursae, and tendon sheaths of the hands and wrists. Our patient with acquired immunodeficiency syndrome and a chronic Prototheca wickerhamii skin infection of the hand developed tenosynovitis and arthritis of his ankle in the setting of a documented algemia. This is the first reported case of protothecal arthritis and tenosynovitis resulting from hematogenous dissemination. The reported musculoskeletal manifestations of protothecal infections are reviewed. PMID- 15338516 TI - Tophaceous pseudotumors in Polynesian patients with gout. PMID- 15338517 TI - Maxillary actinomycosis in an immunocompromised patient with longstanding vasculitis treated with mycophenolate mofetil. PMID- 15338518 TI - Erosions rare without clinically detectable inflammation. PMID- 15338519 TI - Re: GAMES issue. PMID- 15338520 TI - New information for some old editorials. PMID- 15338521 TI - Developments in HIV therapeutics. PMID- 15338522 TI - What's next? Treatment options when the first antiretroviral regimen fails virologically or is not tolerated, Part 1. AB - There are many choices for the initial treatment of HIV infection. However, a change may be required if drug resistance or intolerance occurs. Thus, the selection of an initial regimen is important not only because of its immediate effects but also because of its potential influence on subsequent treatment options. HIV clinicians should be aware of the resistance and safety profiles of the regimens they prescribe as initial therapy and be ready with a strategy for subsequent treatment in the event of virologic failure or drug toxicity. This article suggests treatment algorithms for those experiencing adverse events or the emergence of drug resistance with commonly used antiretroviral regimens. PMID- 15338523 TI - Can antiretroviral therapy ever be stopped? An update. AB - Eradication of virus is not a reasonable goal of anti-HIV therapy because of the latent reservoir of HIV-1 in resting memory CD4+ T cells, which guarantees lifetime persistence of virus. Any drug-resistant viruses that arise and circulate for significant periods can be stored in the reservoir, limiting future treatment options, and mistakes in treatment can be "remembered" by the virus. Wild-type virus is also preserved in the reservoir and can reemerge if therapy is stopped. Since wild-type virus can be more virulent, suppression of this virus is one benefit of continuing therapy in patients who are experiencing treatment failure. If therapy is stopped, the reemergence of wild-type virus does not mean that drug-resistant virus is gone; it is still preserved in the latent reservoir. In patients who have viral suppression to below 50 copies/mL, there is still a low level of virus in the plasma, but it does not appear to be evolving. Thus, in principle, lifelong control of viral replication is possible if suppression of viremia to below 50 copies/mL is maintained. PMID- 15338524 TI - Editorial comment: stopping HAART--where we've been, where we are, where we want to be. PMID- 15338525 TI - Rapid reversal of renal failure after initiation of HAART: a case report. AB - HIV-associated nephropathy (HIVAN), characterized by proteinuria and progressive renal failure, is a well-known complication of HIV infection. Treatment of the condition has focused on the use of corticosteroids, angiotensin-converting enzyme inhibitors, and HAART, all of which can improve the prognosis. Although there are a few case reports of improvement of renal function after the initiation of HAART, this treatment has not been studied in a prospective fashion, and the timing of the improvement is not well documented. Presented here is a case of rapid reversal of end-stage renal failure after the initiation of HAART in a person with biopsy-proven HIVAN. PMID- 15338526 TI - Editorial comment: effect of HAART on HIVAN--what can the clinician expect? PMID- 15338527 TI - Computer assisted bypass management. 1980. PMID- 15338528 TI - A guide to diagnosis of Alzheimer's disease. AB - A major challenge in the management of Alzheimer's disease (AD) is accurate diagnosis of the condition. Aging itself is associated with cognitive change, and physicians must first distinguish between age-associated complaints and true memory impairment as in AD before optimal management can begin. This article reviews tools used to assess and diagnose AD patients in both the primary care and clinical trial settings. AD assessment instruments commonly used in research may be too cumbersome for use in primary care. Tools specifically designed to meet the needs of busy practitioners that can accurately assess function, cognition, and global change will be reviewed. The issues of cognitive screening in old age and age-associated memory impairment will be discussed. Similarly, the pitfalls and advantages of routine cognitive screening in daily practice will be addressed. PMID- 15338529 TI - Proceedings of the Symposium on Novel Therapeutic Opportunities for Neurodegenerative Disease. September 12-13, 2002, Aveiro, Portugal. PMID- 15338530 TI - [Home nursing in the Netherlands since 1950]. AB - This article describes home nursing in the Netherlands between 1950 and 2004. The developments in this period are described form the theoretical perspective on professions of Andrew Abbott: 'professions are exclusive occupational groups applying somewhat abstract knowledge to particular cases'. In 1950, home nursing was an all-round profession providing home nursing care and preventive care to all categories of patients, mainly in their own homes. It was - and still is - a profession situated in the 'periphery' of the health care system, where care and support to patients with pain, suffering and disabilities because of age or chronic illness are considered as belonging to a separate task domain, relatively independent of the mainly curative activities that are performed in the 'medical centre' of health care, especially in the academic hospitals. Typical compared to other countries is that an extensive network of private initiatives, in the form of Cross Organisations of different denominational signatures, existed in the Netherlands until 1990, covering the whole country with home nursing services. In that year, the provision of home nursing and home help were integrated and most home nursing organisations merged into large, regional home care organisations. In this article, six main social developments are described, that influenced the development of home nursing and resulted in the profession as it is now: a differentiated profession divided into different levels of care, working in an organisational, largely bureaucratic setting of home care organisations, where managers and external regional assessment organisations (RIO's) decide on the care to be provided. They now find themselves in a transmural setting, where boundaries between different forms of care no longer exist, and co-operation with other professionals, such as home helps, specialist nurses, GPs, and hospital physicians, is frequent. Currently, their professional autonomy and independent decision-making regarding diagnosis and therapy is affected, and elements of bureaucratisation and managementism (for example aspects such as time-writing) affect their daily work. However, home nursing can still be characterised as a relatively exclusive and independent profession, solving particular cases in the homes of patients by performing activities that are based on abstract, methodical knowledge. PMID- 15338531 TI - [The role of imagination in modern medicine]. AB - In Renaissance and early modern times, the concept of imagination (Latin imaginatio) was essential for the (natural) philosophical explanation of magic processes, especially in the anthropology of Paracelsus. He assumed that imagination was a natural vital power including cosmic, mental, phychical, and physical dimensions. The Paracelsians criticized traditional humor pathology ignoring their theory of' 'natural magic'. On the other hand, they were criticized by their adversaries as charlatans practicing 'black magic'. About 1800, in between enlightenment and romanticism, the healing concept of, animal magnetism' (Mesmerism) evoked an analogous debate, whether, magnetic' phenomena originated from a real (physical) power (so-called, fluidum') or were just due to fantasy or imagination (German Einbildungskraft). At the end of the 19th century, the French internist Hippolyte Bernheim created-against the background of medical hypnosis (hypnotism') as a consequence of Mesmerism - his theory of suggestion and autosuggestion: a new paradigm of psychological respectively psychosomatic medicine, which became the basis for the concept of, placebo' in modern biomedicine. From now on, all the effects of, alternative medicine' could easily be explained by the, placebo-effect', more or less founded - at least unconsciously - on fraud. PMID- 15338532 TI - ExTech 2003. Proceedings of the 5th International Symposium on Advances in Extraction Technologies. St. Pete Beach, Florida, USA, 5-7 March 2003. PMID- 15338533 TI - Transporting patient with suspected SARS. PMID- 15338534 TI - Keeping tabs. Benchmark data show overall department trends. PMID- 15338535 TI - [Semantics of learned quackery in the 17th and 18th centuries]. AB - In the 17th and 18th century republic of letters the problem of scientific fraud was met with a discourse of charlatanism. Departing from Johann Burchhard Menckes famous treatise on the Charlatanry of the learned the following essay traces how the accusations of academic and scientific misconduct put in terms of 'charlatanry' primarily helped to produce the new species of the erudite 'charlatan'. Facing a growing complexity of scientific culture this new frame of meaning, structured by numerous examples of scientific misconduct offered a new way of orientation in the world of learning. But besides its cognitive impacts the discourse of charlatanry allowed to create symbolic boundaries, which determined decisions upon the affiliation or non affiliation to the new forming scientific community by separating honourable from dishonourable scientific personae. Speaking of charlatanry therefore always implied a social distinction as much as a scientific. The discourses on charlatanry also mirror differentiations within the scientific field. At first dominated by a critique built on courteous or bourgeois values, the scientific field later on developed its own criteria of appraisal like authorship, originality, transparency etc. Attracting the attention of a further growing public sphere, the explicit verbalisation of claims not relating to the value system of a republic of letters primarily concerned with the production and distribution of knowledge finally led up to a more implicit moral economy of science. A change that at a large scale level can be described both as an internalisation of the values of scientific conduct and differentiation between justiciable and unjusticiable transgressions of the norms set up by the scientific community. PMID- 15338536 TI - Psychosocial impact of SARS. PMID- 15338537 TI - Highly pathogenic avian flu, Japan. PMID- 15338538 TI - Alexander the Great and West Nile virus encephalitis. PMID- 15338539 TI - Alexander the Great and West Nile virus encephalitis. PMID- 15338540 TI - Alexander the Great and West Nile virus encephalitis. PMID- 15338541 TI - Syndromic surveillance. PMID- 15338542 TI - Staphylococcus aureus and Escherichia hermanii in diabetes patient. PMID- 15338543 TI - What is the history of medieval optics really about? PMID- 15338544 TI - Paradoxes of free will and the limits of human reason. PMID- 15338545 TI - Multiple rpoB mutants of Mycobacterium tuberculosis and second-order selection. PMID- 15338546 TI - Human metapneumovirus and chronic obstructive pulmonary disease. PMID- 15338547 TI - Pathologic quiz case. Edema and diarrhea in a 2-year-old boy. Menetrier disease with cytomegalovirus gastritis. PMID- 15338548 TI - Integrons in Salmonella Keurmassar, Senegal. PMID- 15338549 TI - Hepatitis B infection, Eastern India. PMID- 15338550 TI - Cardiovascular medicine and surgery in Singapore: the next lap. PMID- 15338551 TI - Ehrlichia prevalence in Amblyomma americanum, Central Texas. PMID- 15338552 TI - Echinococcus multilocularis in Northern Hungary. PMID- 15338553 TI - Antimicrobial resistance in Campylobacter. PMID- 15338554 TI - Progression of chronic myeloid leukemia to blast crisis during treatment with imatinib mesylate. AB - CONTEXT: Previous investigators have reported discrepancies between hematologic, marrow morphologic, and cytogenetic responses to imatinib mesylate among patients with chronic myeloid leukemia (CML). In addition to disease refractoriness, rare instances of disease progression from chronic phase to blast crisis during imatinib therapy have recently been anecdotally reported. OBJECTIVES: To describe the clinicopathologic features of 3 patients with CML who rapidly progressed from chronic phase to blast crisis while taking imatinib and to perform a review of the literature. DESIGN: Morphologic, immunophenotypic, and cytogenetic analyses were performed on the 3 patients at the time of initial diagnosis, during imatinib therapy, and at blast crisis. RESULTS: The 3 patients were men, aged 39, 42, and 43 years. Two had been treated with hydroxyurea for 16 and 21 months before imatinib therapy, while 1 was started on a regimen of imatinib following diagnosis. Despite a hematologic response in all 3 patients, none of them achieved cytogenetic remission, and all progressed to blast crisis at 7 to 10 months of imatinib therapy. Blood findings during blast transformation were heterogeneous, including normal blood morphologic findings in 1 patient, leukocytosis with circulating blasts and basophilia in 1, and marked pancytopenia in 1. All 3 marrow specimens demonstrated moderate to marked diffuse reticulin fibrosis with more than 20% blasts. Clonal cytogenetic evolution was evident in 2 of the 3 patients and included an extra Philadelphia chromosome in both. All 3 patients underwent allogeneic bone marrow transplantation. One was alive with no evidence of disease at 14 month follow-up, while 2 had residual disease after bone marrow transplantation and died of complications at 4 and 5 months after transplantation. CONCLUSIONS: Blood data did not always reflect marrow status. Therefore, bone marrow follow-up is critical for monitoring of response. Our findings suggest that significant progression of marrow reticulin fibrosis during imatinib therapy can be an indicator for a return or progression of CML and, in some patients with CML, imatinib may promote cytogenetic clonal evolution, resulting in a poor response to treatment. PMID- 15338555 TI - [Consensus Development Conference on Thrombosis and Pregnancy. 14 March 2003, Paris, France. Proceedings]. PMID- 15338556 TI - Proceedings of the 55th International Symposium on Crop Protection. Gent, May 6, 2003. PMID- 15338557 TI - Medicaid program information. PMID- 15338558 TI - A hydatidiform mole in a postmenopausal woman. A case report and review of the literature. AB - Gestational trophoblastic disease occurs in less than 1 per 1200 pregnancies in the United States. The spectrum of this disease ranges from benign hydatidiform mole to trophoblastic malignancy (placental-site trophoblastic tumor and choriocarcinoma). Benign gestational trophoblastic disease generally occurs in women of reproductive age and is extremely rare in postmenopausal women. To our knowledge, our case represents only the third description in the world literature of a benign complete hydatidiform mole in a woman with a history of amenorrhea greater than 1 year. We describe the case of a 61-year-old postmenopausal woman who underwent an emergent total abdominal hysterectomy due to uncontrollable vaginal bleeding associated with an increased serum beta-human chorionic gonadotropin level. The resected uterus contained an endometrial, cystic, grapelike tumor. Microscopic examination demonstrated hydropic degenerated villi with a circumferential trophoblastic cell proliferation and moderate atypia, consistent with a complete hydatidiform mole. The morphologic and immunophenotypic characteristics are presented, as well as the results of a literature review. PMID- 15338559 TI - An isolated vertebral metastasis of urachal adenocarcinoma. AB - We describe a case of urachal adenocarcinoma arising in a 30-year-old woman. The patient underwent partial cystectomy. Three years later, she presented with an isolated metastasis to a thoracic vertebra, which was treated by vertebral corporectomy. Histologic findings are described, and the clinical findings, management, and metastatic patterns of this rare tumor, as well as the differential diagnosis on a biopsy specimen, are discussed. PMID- 15338560 TI - Sclerosing mucoepidermoid carcinoma of the parotid gland. AB - Although mucoepidermoid carcinoma is the most common primary malignancy of the salivary glands, the sclerosing morphologic variant of this tumor is extremely rare, with only 6 reported cases. As its name suggests, sclerosing mucoepidermoid carcinoma is characterized by an intense central sclerosis that occupies the entirety of an otherwise typical tumor, frequently with an inflammatory infiltrate of plasma cells, eosinophils, and/or lymphocytes at its peripheral regions. The sclerosis associated with these tumors may obscure their typical morphologic features and result in diagnostic difficulties. Tumor infarction and extravasation of mucin eventuating in reactive fibrosis are 2 mechanisms of formation that have been suggested as underlying this morphologic variant. We describe herein another case of sclerosing mucoepidermoid carcinoma that was diagnosed in a 44-year-old woman and review the relevant literature. Morphologic evidence in support of the mucin extravasation hypothesis was identified, as small pools of mucin were present throughout the tumor. However, there was no concentration of the mucin pools near the areas with the most viable tumor cells, which would have provided evidence for a temporal sequence that eventuates in lack of mucin in the most sclerotic regions. PMID- 15338561 TI - Pathologic quiz case. A 29-year-old man with a tongue lesion. Granular cell tumor. PMID- 15338562 TI - Erwin Chargaff. PMID- 15338563 TI - Gender differences in cortical complexity. AB - Cortical complexity, a measure that quantifies the spatial frequency of gyrification and fissuration of the brain surface, has not been thoroughly characterized with respect to gender differences in the human brain. Using a new three-dimensional (3D) analytic technique with magnetic resonance imaging, we found greater gyrification in women than men in frontal and parietal regions. Increased complexity implies more cortical surface area, which may offset gender differences in brain volume and account for behavioral gender differences. PMID- 15338564 TI - Max Perutz. PMID- 15338565 TI - Little by little. PMID- 15338566 TI - Profile: Sharon Mitchell, head of the Adult Industry Medical Clinic. PMID- 15338567 TI - Paul H Silverman. PMID- 15338568 TI - Evidence that xeroderma pigmentosum cells do not perform the first step in the repair of ultraviolet damage to their DNA. 1969. AB - Xeroderma pigmentosum (XP) is a recessively transmitted disorder of man characterized by increased sensitivity to ultraviolet light. Homozygous, affected individuals, upon exposure to sunlight, sustain severe damage to the skin; this damage is characteristically followed by multiple basal and squamous cell carcinomas and not uncommonly by other malignant neoplasia. A tissue culture cell line was derived from the skin of a man with XP. Our measurements of ultraviolet induced pyrimidine dimers in cellular DNA show that normal diploid human skin fibroblasts excise up to 70 per cent of the dimers 24 hours, but that fibroblasts derived from the individual with XP excise less than 20 per cent in 48 hours. Alkaline gradient sedimentation experiments show that during the 24 hours after irradiation of normal cells a large number of single-stranded breaks appear and then disappear. Such changes are not observed in XP cells. XP cells apparently fail to start, the excision process because they lack the required function of an ultraviolet-specific endonuclease. These findings, plus earlier ones of Cleaver on the lack of repair replication in XP cells, raise the possibility that unexcised pyrimidine dimers can be implicated in the oncogenicity of ultraviolet radiation. PMID- 15338569 TI - Emerging infections: what have we learned from SARS? PMID- 15338570 TI - Open access publishing. PMID- 15338571 TI - The chemical hunt for the identification of drugable targets. AB - Chemical biology has emerged as a new scientific discipline to change the way scientists approach and study the interface between chemistry, biology, and physics. By integrating the knowledge base of the human genome with the power of diverse and flexible chemical technology platforms, the ultimate goal is to define the 'rules of engagement' for small molecules and their use in basic biology and in drug discovery. Herein, we highlight the current counterpoles of the chemical biology philosophy in the framework between conformational diversity and informational complexity. Expanding the growing molecular recognition information matrix into classification of diseases and immediate mechanistic in vivo proof of concept models represent the next development phase in a field that, unlike any other due to its multidisciplinary nature, unifies basic scientists and drug discoverers. PMID- 15338572 TI - The seventh annual meeting of the American Society of Gene Therapy. PMID- 15338573 TI - [When the Egyptian mummies are speaking about the infections that have made them ill]. AB - The microbiological study of mummies has started in 1910 when Sir M.A. Ruffer first applied the histologic methods to the study of mummified tissues and found Schistosoma haematobium ovas dated from the XXth dynasty. Up to the 1990 years, morphological methods including radiology, computed tomography, endoscopy, history, electron microscopy, and serology have been the main tools used in Paleopathology. They led to identify schistosomiasis, dracunculiasis, trichinosis, ascariasis and bone tuberculosis as the most prevalent diseases of the ancient residents of Egypt. The recent introduction of molecular methods (PCR) allowed t confirm the high prevalence of helminth diseases and tuberculosis among these populations, but also added new data exemplified by the widespread distribution of Plasmodium falciparum malaria. In addition, cases of bacterial septicemias and diphteria possibily occurred. Thousands of human and animal mummies remain to be studied with the hope to discover another pathogens responsible for viral or zoonotic infections prevalent during the pharaons' times. PMID- 15338574 TI - THTA, a thermotolerance gene of Aspergillus fumigatus. AB - Aspergillus fumigatus grows optimally from 37 to 42 degrees C but can grow at temperatures up to 55 degrees C. To study the genetic basis of thermotolerance and its role in virulence of A. fumigatus, temperature sensitive mutants were isolated. One of the mutants that grew at 42 degrees C but not at 48 degrees C was complemented and the gene, THTA, was identified. Deletion of THTA showed the same temperature sensitivity as the original mutant. THTA encodes a putative protein of 141 kDa with unknown function and the HA-tagged ThtAp accumulated to similar levels in cultures grown at either 37 or 48 degrees C. Southern blot analysis and database searches revealed the presence of THTA-related sequences in several other ascomycetous fungi. No difference in virulence was observed between the deltathtA and wild-type strains. Thus, THTA is essential for growth of A. fumigatus at high temperatures but does not contribute to the pathogenicity of the species. PMID- 15338575 TI - Trends in Americans' access to needed medical care, 2001-2003. AB - Despite sluggish economic growth and rapidly rising health care costs, Americans' access to needed medical care improved between 2001 and 2003, especially among low-income children and adults, according to findings from the Center for Studying Health System Change's (HSC) nationally representative Community Tracking Study Household Survey. In particular, the proportion of low-income, uninsured Americans who reported going without needed medical care fell by 3.2 percentage points to 13.2 percent in 2003, and unmet medical needs for low-income children decreased to the point where income-related differences in access to care for children have disappeared. Nonetheless, about one in seven Americans reported difficulty obtaining needed care in 2003, and people reporting access problems increasingly cited cost as a barrier to care. PMID- 15338576 TI - Advanced glycation end products in urine: are some sugar-derived products better than others? PMID- 15338577 TI - Current awareness in human psychopharmacology. PMID- 15338578 TI - A weighty issue for children. PMID- 15338579 TI - [Nutritional status of Mexican women]. AB - The available epidemiological information of nutrition status of Mexican women at reproductive age is presented. The consulted sources were The National Nutrition Survey and The National Survey of Chronic Diseases and three local surveys performed in open populations (Urban Nutrition Survey), one performed in San Luis Potosi, one in Merida, Yucatan and one performed in a specific zone in Mexico City (delegacion Alvaro Obregon). All surveys included the evaluation of women at reproductive age and registered in a direct form weight and height to calculate the body mass index (BMI), however, they did not use the same criteria to define low weight, overweight and obesity. Even with these limitations, the different studies showed that while to some populations stratums, problems related to insufficient feeding persist (close to 29% with a BMI < 20.1), according to The National Nutrition Survey), for most of the women in reproductive age overweight and obesity are a serious health problem (40.7% of women between 20 to 49 years old have a BMI > or = 27.3), according to The National Survey of Chronic Diseases. Information of the risk increment to present hypertension and Diabetes Mellitus in overweight and obese women is shown. PMID- 15338580 TI - [Non-pregnant women's nutrition and its impact in life quality]. AB - Emphasis is made in the nutrition aspects related to women at reproductive age that are not pregnant or lactating and that includes the variations that happen throughout the menstrual cycle, fluctuations in energy expenditure, body composition and mood. Nutrition role in some premenstrual syndrome alterations as premenstrual stress (serotonin, magnesium, calcium and vitamin E), anemia, gynecological cancers (antioxidants, alcohol, folic acid, lipids, fiber and phytosterols) and osteoporosis (exercise and diet) are also described, as well as the impact on nutrition of the use of contraceptive methods (hormonal and intrauterine devices). Practical recommendations directed toward the evaluation and management of the main nutrition needs of adult women are included. PMID- 15338581 TI - [Maternal nutrition effect on fetus development and pregnant women's health]. AB - A review is made of the main prenatal growth stages and the way they are influenced by social and biological factors pointing out the role of maternal nutrition before and during pregnancy. It is discussed the available evidence on the relation of maternal nutrition and fetal growth through clinical and epidemiological studies performed in animal and humans, analyzing the causal mechanisms of this association, including the role of placenta. Some clinical useful indicators that allow to evaluate maternal nutrition throughout gestation are presented. Emphasis is made on the impact of protein-energy malnutrition as well as on some specific deficiencies, particularly iron and folic acid. At last, the synergic effect of infection and malnutrition on low birth weight and prematurity is analyzed. PMID- 15338582 TI - [Nutrition status evaluation of the women at reproductive age. Uses and limitations]. AB - Information is given about the main methods for nutrition status evaluation of the woman, this information is oriented to three stages of the woman's life cycle: adult stage, gestation stage and lactation stage. The usefulness as well as the limitations of the diet surveys are discussed. The main anthropometric indicators are presented, useful in the clinical practice to evaluate nutrition status of the non-pregnant woman (body mass index, waist/hip index and skinfolds), pregnant woman (percentage of expected weight for height and gestation age, weight gain during pregnancy, height of fundus of uterus, arm circumference and skinfolds) and lactating woman (weight loss and skinfolds). The most important nutrition status biochemical indicators are discussed. At last some special evaluation technics as measurements of body composition and balance methods are discussed. PMID- 15338583 TI - [Dietary recommendations of women in reproductive age]. AB - The characteristics of an adequate diet are described and defined. A simple method for the calculation of energy needs of adult, pregnant and lactating women is described. A food exchange list is presented, based on 8 food groups, including ration sizes of every one of them. Group I: cereals and tubercles; Group II: vegetables (according to their energy supply); Group III: fruits (according to their fiber content); Group IV: legumes; Group V: meats and egg (classify according to their lipid content); Group VI: dairy products (divided according to their lipid content); Group VII: lipids (fats and oils) and Group VIII: sugars. The main sources of critical nutrients for women at reproductive age are pointed out, as calcium, iron, folic acid, vitamin C, vitamin B12, vitamin A and carotenes. Some menus are described to illustrate the use of the food exchange list. PMID- 15338584 TI - Stress protein assay for the evaluation of cytotoxicity of dental amalgam. AB - To evaluate the cytotoxicity of mercury in dental amalgams, a stress protein assay was performed and the results were compared with the cytotoxicity evaluated by a neutral red uptake assay. The induction of a major stress protein, hsp70, was analyzed at levels of mRNA, synthesis and accumulation in human HeLa cells treated with extracts from amalgam, metal mercury and mercuric chloride. Mercuric chloride induced an increase in the synthesis of hsp70 at concentrations of mercury half those used for the neutral red uptake assay. The extracts from dental amalgam and metal mercury induced an increase in hsp70 mRNA at concentrations of mercury half those causing the inhibition of neutral red uptake into cells. Furthermore, the extracts from dental amalgam or metal mercury increased the synthesis of hsp70 and inhibited the uptake of dye at concentrations of mercury 1/10-1/50 lower than those at which mercuric chloride acted. These results suggest that the stress protein assay is more sensitive than the conventional neutral red assay for the evaluation of the cytotoxicity of mercury in dental amalgams and that the methods used in the preparation of metal solutions seem to be critical to the evaluation of cytotoxicity of dental materials. PMID- 15338585 TI - Flexural properties of glass fiber reinforced composite with multiphase biopolymer matrix. AB - The aim of this study was to evaluate flexural properties of glass fiber reinforced composites with a multiphase biopolymer matrix. Continuous unidirectional E-glass fibers were preimpregnated with a novel biopolymer of poly(hydroxyproline) amide and ester. The preimpregnated fibers were then further impregnated in a co-monomer system of Bis-GMA-TEGDMA, which formed semi interpenetrating polymer networks (semi-IPN) with the preimpregnated polymer. After light initiated polymerization of the monomer system, rectangular shaped bar specimens (n = 4) were tested by the three-point bending test. The control material was a fiber-reinforced composite with a Bis-GMA-TEDGMA-matrix only. The mean flexural strength of poly(hydroxyproline) amide preimpregnated fiber composite was higher than that of the control (FS = 888 vs. 805 MPa). The poly(hydroxyproline) ester preimpregnated fibers resulted in lower strength (FS = 541 MPa). The results of this study suggest that preimpregnation of glass fibers with poly(hydroxyproline) amide and the use of such fibers in fiber-reinforced composites with IPN polymer matrices, can reach relatively high mechanical properties. PMID- 15338586 TI - Basic properties of apatite cement containing spherical tetracalcium phosphate made with plasma melting method. AB - Apatite cement (AC) can be injected through syringe and forms apatite mass. Therefore, AC is very useful for a minimally invasive surgical operation aimed for the reconstruction of bone defects. However, injectability of current AC is not satisfactory for its clinical use. In this investigation, therefore, spherical tetracalcium phosphate (s-TTCP) was prepared with plasma melting method and its effect on injectability were evaluated as well as other basic properties of AC. We found much better handling property and injectability when we used s TTCP as a component of AC (s-AC). For example, cement spread area used as an index of consistency of the s-AC paste was 512 mm2 whereas that of ordinary AC with irregular TTCP (i-AC) was 158 mm2 when powder to liquid mixing ratio was 2.5. However, diametral tensile strength of set s-AC (1.4 MPa) was significantly lower than that of set i-AC (10.7 MPa) when the powder to liquid ratio was 4.0. X ray powder diffraction analysis revealed limited formation of apatite in the case of s-AC. Although there are some drawbacks, we feel the use of spherical particle is very useful to improve the injectability of AC. Therefore, it is important to find suitable method to prepare spherical powder as the component of AC. PMID- 15338587 TI - Nano-scaled hydroxyapatite/polymer composite I. Coating of sintered hydroxyapatite particles on poly(gamma-methacryloxypropyl trimethoxysilane)grafted silk fibroin fibers through chemical bonding. AB - The inorganic-organic composite consisting of nano-scaled hydroxyapatite (HAp) and silk fibroin (SF) fibers was prepared through covalent linkage to develop a novel biomaterial for a soft-tissue-compatible material. The preparation of the composite was conducted through the three-step procedure consisting of chemical modification using 2-methacryloxyethyl isocyanate (MOI) monomer to introduce vinyl groups on SF, poly(gamma-methacryloxypropyl trimethoxysilane) (MPTS) graft polymerization on SF, and coupling process between the surface of polyMPTS grafted SF and HAp nano-particles. The amount of the graft-polymerization of polyMPTS through vinyl groups was well controlled by the reaction time. The nano crystals were subsequently coated on the grafted fibers by heating at 120 degrees C for 2 h in a vacuum. The crystalline structure of the SF substrate did not change in the procedure. In the SEM observation of the composite surface, it was found that the bonded nano-crystals were separated and partially aggregated with several crystals attached on the SF fiber surface. The HAp particles adhered more strongly on the SF surface with separation or aggregation of several crystals than on the surface of the original SF after ultrasonic treatment. PMID- 15338588 TI - The influence of lithium fluoride on in vitro biocompatibility and bioactivity of calcium aluminate-pMMA composite cement. AB - The objective of this study is to assess the influence of lithium fluoride on in vitro biocompatibility and bioactivity of calcium aluminate (CA) polymethylmethacrylate (PMMA) composite cement exhibiting quick setting time ( < 15 min), low exothermic temperature (< 47 degrees C), and high compressive strength (> 100 MPa). The biocompatibility was measured by examining cytotoxicity tests such as the agar diffusion test with L929 cell line and the hemolysis test with fresh rabbit blood. To estimate the bioactivity of CA-PMMA composite cement, we determined hydroxyapatite (HAp) formation on the surface of composite cement in the simulated body (SBF) solution by using thin-film XRD, XPS, SEM, EPMA and ICP-AES. The results of biocompatibility tests indicated that all experimental compositions of this study had no cytotoxicity and no hemolysis so that there was no cytotoxicity with regard to non-reacted monomers (MMA and TEGDMA) and lithium fluoride. The results of bioactivity tests revealed that CA-PMMA composite cement without lithium fluoride did not form HAp on its surface after 60 days of soaking in the SBF. On the other hand, LiAl2(OH)7 . 2H2O and HAp were formed on the surface of CA-PMMA composite cement including 1.0% by weight of lithium fluoride after 7 and 15 days of soaking in the SBF, respectively. The 5 microm of LiAl2(OH)7 . 2H2O and HAp mixed layers were formed on the surface of specimen after 60 days of soaking in the SBF. PMID- 15338589 TI - Modification of human platelet adhesion on biomaterial surfaces by protein preadsorption under static and flow conditions. AB - Biomaterial-induced thrombosis remains one of the main complications of vascular implant devices. Preadsorbed proteins on the biomaterial/blood interface will modify the adhesion and activation of platelets (PTLs) during the initial contact phase. Our results clearly show that PTL-adherence on biomaterials is influenced not only by protein preadsorption, but also by flow conditions. The covalent coating of TCPS and glass by phosphorylcholine (PC) induces a significant decrease of PTL adhesion but leads to a slight, but nevertheless significant activation of PTL, which was detected by the induction of P-selectin expression using FACS analysis. Methodologically, the visualization of PTL adhesion gave more reliable results for measurement of PTL adhesion than the cell-enzyme immunoassay (EIA) for P-selectin. Human citrated plasma caused an inhibition of PTL. It is probable, that the contained sodium citrate may inhibit PTL adhesion by its calcium ion-binding capacity. The flow experiment as dynamic system is in our view absolutely essential for the evaluation of biomaterials for vascular prosthesis, and is in accordance with the international standards. The results of the experiments also suggest that investigations under static and flow conditions are needed to determine the influence of protein adsorption on mixed blood cell populations, for example, on PTL and PMN mixtures/co-cultures in order to achieve a better simulation of the in vivo situation. PMID- 15338590 TI - In vitro release dynamics of insulin from a loaded hydrophilic polymeric network. AB - A hydrophilic semi-interpenetrating polymer network of polyvinyl alcohol (PVA), poly(ethylene glycol) (PEG) and crosslinked polyacrylamide (PAM) chains has been synthesized and its potential for controlled release of macromolecular drugs has been assessed by taking insulin as a representative drug. The semi-IPN was characterized by IR studies and network parameters such as the average molecular weight between crosslinks (Mc), crosslink density (q), and number of elastically effective chains (Ve) were evaluated. The effect of chemical architecture of the IPN was investigated on the percent loading of insulin and its subsequent release from the loaded device. Other parameters such as the thickness of the gel, molecular weight of PEG and pH and temperature of the release medium were also studied for their possible impact on the release of insulin. The whole release data was analyzed by Ficks power law and the influence of various factors on the plausible mechanism of insulin release was investigated. PMID- 15338591 TI - Electrochemical impedance spectroscopy characterization of passive film formed on implant Ti-6Al-7Nb alloy in Hank's solution. AB - The influence of potential on electrochemical behavior of Ti-6Al-7Nb alloy under simulated physiological conditions was investigated by electrochemical impedance spectroscopy (EIS). The experimental results were compared with those obtained by potentiodynamic polarization curves. All measurements were carried out in Hank's aerated solution at 25 degrees C, at pH 7.8 and at different potentials (corrosion potential, 0 mV(SCE), 1000 mV(SCE), and 2000 mV(SCE)). The EIS spectra exhibited a two-step or a two-time constant system, suggesting the formation of a two-layer oxide film on the metal surface. The high corrosion resistance, displayed by this alloy in electrochemical polarization tests, is due to the dense inner layer, while its osseointegration ability can be ascribed to the presence of the outer porous layer. PMID- 15338592 TI - Fatigue characterization of a polymer foam to use as a cancellous bone analog material in the assessment of orthopaedic devices. AB - Analog materials are used as a substitute to cancellous bone for in vitro biomechanical tests due to their uniformity, consistency in properties and availability. To date, only the static material properties of these materials have been assessed, although they are often used in fatigue tests. Cancellous bone exhibits complex material behavior when subjected to fatigue loads, including modulus degradation, accumulation of permanent strain and increasing hysteresis. Analog materials should exhibit similar fatigue behavior to cancellous bone if they are to be used in cyclic loading tests. In our study, a polymer foam (commercial name HEREX C70.55) has been studied for its static and fatigue behavior and compared with that of cancellous bone. In compression, the foam exhibited qualitatively similar mechanical behavior, but the degree of modulus degradation and accumulation of permanent strain was lower than expected for cancellous bone. In general, the tensile properties of the foam were greater than found in compression, the opposite to the mechanical behavior of cancellous bone. The methodology employed here could form the basis of selecting suitable analog materials for cancellous bone in the future. PMID- 15338593 TI - In vitro cytotoxicity of E-glass fiber weave preimpregnated with novel biopolymer. AB - The aim of this study was to investigate cytotoxicity of composition of E-glass fibers and novel biopolymer of poly(hydroxyproline). Growth and proliferation of the human gingival fibroblast cells on the surface of the materials was evaluated. The number of cells grown and proliferated on cell culture plastic was used as a control. Bi-directional fiber weaves were preimpregnated with poly(hydroxyproline). Cytotoxicities of the preimpregnated and nonimpregnated materials were evaluated bythe release of lactate dehydrogenase from the cells during the culture period of 24 h. The values of the lactate dehydrogenase activity of the materials' extracts showed non-toxicity for poly(hydroxyproline) preimpregnated E-glass fiber weaves. The growth of fibroblasts on the surface of the materials appeared normal after 11 days culture period; they looked healthy and normal in size and shape. The results of this study suggest that based on its' non-cytotoxicity the composition of E-glass fibers and poly(hydroxyproline) can further be evaluated as a material that is suitable for biomedical use. PMID- 15338594 TI - Optimization of the formulation and mechanical properties of starch based partially degradable bone cements. AB - Previous studies have shown the possibility of developing in-situ polymerizable hydrogels that disclose a range of properties that might allow for their use as bone cements. Their main advantage is to be partially degradable, which is important to allow bone ingrowth (better fixation). In addition, their uptake of water makes them less agressive for the tissues, disclosing better fatigue properties and ideal for release of drugs when in service. This work reports a statistical study of the formulation of partially degradable acrylic bone cements that include on their composition corn starch/cellulose acetate blends (SCA). The aim was to optimize a set of properties (mechanical, swelling/degradation and curing) by changing the values of some parameters such as SCA amount and particle size and molar ratio of the acrylic monomers. Statistical tests demonstrated that the most important parameter was the molar ratio of monomers, with the SCA percentage also playing a role. It was possible to develop formulations with mechanical properties in the range of ASTM specifications and with polymerization temperatures lower than those of commercial acrylic cements. Some formulations were subsequently selected for tensile and dynamic mechanical thermal analysis (DMA) tests, under dry and wet conditions. PMID- 15338595 TI - Thermal characterization of PMMA-based bone cement curing. AB - In thermal characterization tests of polymethylmethacrylate bone cement performed according to the ASTM Standard Specification for Acrylic Bone Cement, time temperature profiles of bone cement were observed to be sensitive to the thickness of the cement patty and the mold material. Due to the heat transfer from cement to the surrounding mold, such tests might underestimate the exothermic temperature of bone cement. Developing test methods to better characterize cement thermal behavior is necessary for accurate cement curing simulations. In this paper, the effects of the mold material and geometry on experimental measurements of bone cement setting temperature and setting time were evaluated by conducting the polymerization in different test molds. Finite element (FE) numerical simulations were also performed to provide a further understanding of these effects. It was found that the mold material and geometry significantly influence the values of the parameters measured using the ASTM standard. Results showed that the setting temperature measured was about 50 degrees C lower in a polytetrafluoroethylene (PTFE) mold than in a polyurethane (PU) foam mold for the 6 mm thickness cement. The measured peak temperature using PTFE molds varied about 75 degrees C for different mold heights (6mm vs. 40 mm), but only by 28 degrees C with PU molds. The measured setting time with PTFE molds varied by about 740 s for different mold heights (6 mm vs. 40 mm), while only by about 130 s for PU molds. Using PU foam materials for the test mold decreases cement heat transfer effects due to the poor heat conductivity of PU foam and provides more consistent measured results. FE parametric studies also support these observations. Poor conductivity materials, like PU foam, make better molds for the characterization of bone cement thermal behavior. PMID- 15338596 TI - Carbon-carbon composite bearing materials in hip arthroplasty: analysis of wear and biological response to wear debris. AB - Ultra-high molecular weight polyethylene wear particles have been implicated as the major cause of osteolysis, implant loosening and late aseptic failure in total hip arthroplasties in vivo. This study initially screened 22 carbon-carbon composite materials as alternatives for UHMWPE in joint bearings. New bearing materials should satisfy certain criteria--they should have good wear properties that at least match UHMWPE, and produce wear particles with low levels of cytotoxic and osteolytic activity. Initial screening was based on wear resistance determined in short-term tribological pin-on-plate tests. Three materials (HMU PP(s), HMU-RC-P(s), and SMS-RC-P(s)) which had superior wear resistance were selected for long-term testing. All materials had very low wear factors and SMS RC-P(s), which had a wear factor of 0.08 +/- 0.56 x 10(-7) mm3/Nm, was selected for the subsequent biological testing and particle size analysis. SMS-RC-P(s) showed good biocompatibility in bulk material form and also the wear particles had low cytotoxicity for L929 fibroblasts in culture compared to metal wear particles. Wear debris size analysis by transmission electron microscopy showed that the particles were very small, with the vast majority being under 100 nm in size, similar to metal wear particles. The potential osteolytic effect of SMS-RC P(s) wear particles was investigated by culturing particles with human peripheral blood mononuclear cells and measuring TNFalpha production. SMS-RC-P(s) did not significantly stimulate TNFalpha production at a particle volume to cell number ratio of 80:1, indicating that the debris had a low osteolytic potential. The results of this study suggest that carbon-carbon composites, particularly those composed of PAN-based fibers may be important biomaterials in the development of next generation bearing surfaces for use in total joint replacements that have very low wear rates and reduced osteolytic and cytotoxic potential. PMID- 15338597 TI - The finished genome sequence of Homo sapiens. PMID- 15338598 TI - The human genome: genes, pseudogenes, and variation on chromosome 7. PMID- 15338599 TI - Mutational profiling in the human genome. PMID- 15338600 TI - Assessing the quality of finished genomic sequence. PMID- 15338601 TI - Human subtelomeric DNA. PMID- 15338602 TI - Genome research: the next generation. PMID- 15338603 TI - DNA sequence variation of Homo sapiens. PMID- 15338604 TI - SNP genotyping and molecular haplotyping of DNA pools. PMID- 15338605 TI - Highly parallel SNP genotyping. PMID- 15338606 TI - Structure of linkage disequilibrium in humans: genome factors and population stratification. PMID- 15338607 TI - Genome-wide association of haplotype markers to gene expression levels. PMID- 15338608 TI - Genetic variation and the control of transcription. PMID- 15338609 TI - Genome-wide detection and analysis of recent segmental duplications within mammalian organisms. PMID- 15338610 TI - The effects of evolutionary distance on TWINSCAN, an algorithm for pair-wise comparative gene prediction. PMID- 15338611 TI - Lineage-specific expansion of KRAB zinc-finger transcription factor genes: implications for the evolution of vertebrate regulatory networks. PMID- 15338612 TI - Sequence organization and functional annotation of human centromeres. PMID- 15338613 TI - Evolutionary strategies of human pathogens. PMID- 15338614 TI - Gene expression profiling of cells, tissues, and developmental stages of the nematode C. elegans. PMID- 15338615 TI - Building comparative maps using 1.5x sequence coverage: human chromosome 1p and the canine genome. PMID- 15338616 TI - Positional identification of structural and regulatory quantitative trait nucleotides in domestic animal species. PMID- 15338617 TI - Evolving methods for the assembly of large genomes. PMID- 15338618 TI - Mouse Genome Encyclopedia Project. PMID- 15338619 TI - DNA sequence assembly and multiple sequence alignment by an Eulerian path approach. PMID- 15338620 TI - Ensembl: a genome infrastructure. PMID- 15338621 TI - Prediction, annotation, and analysis of human promoters. PMID- 15338622 TI - Ontologies for biologists: a community model for the annotation of genomic data. PMID- 15338623 TI - The Genome Knowledgebase: a resource for biologists and bioinformaticists. PMID- 15338624 TI - The share of human genomic DNA under selection estimated from human-mouse genomic alignments. PMID- 15338625 TI - Detecting highly conserved regions of the human genome by multispecies sequence comparisons. PMID- 15338626 TI - Comparative analysis of human chromosome 22q11.1-q12.3 with syntenic regions in the chimpanzee, baboon, bovine, mouse, pufferfish, and zebrafish genomes. PMID- 15338627 TI - Genome-wide analyses based on comparative genomics. PMID- 15338628 TI - Comparative genomic tools for exploring the human genome. PMID- 15338629 TI - Evolution of eukaryotic gene repertoire and gene structure: discovering the unexpected dynamics of genome evolution. PMID- 15338630 TI - Human-mouse comparative genomics: successes and failures to reveal functional regions of the human genome. PMID- 15338631 TI - High-throughput mouse knockouts provide a functional analysis of the genome. PMID- 15338632 TI - Identification of novel functional elements in the human genome. PMID- 15338633 TI - High-resolution human genome scanning using whole-genome BAC arrays. PMID- 15338634 TI - Annotation of novel proteins utilizing a functional genome shotgun coupled with high-throughput protein interaction mapping. PMID- 15338635 TI - Global predictions and tests of erythroid regulatory regions. PMID- 15338636 TI - Systems approaches applied to the study of Saccharomyces cerevisiae and Halobacterium sp. PMID- 15338637 TI - Implications of genomics for public health: the role of genetic epidemiology. PMID- 15338638 TI - A model system for identifying genes underlying complex traits. PMID- 15338639 TI - Genomic variation in multigenic traits: Hirschsprung disease. PMID- 15338640 TI - Genetics of schizophrenia and bipolar affective disorder: strategies to identify candidate genes. PMID- 15338641 TI - The genetics of common diseases: 10 million times as hard. PMID- 15338642 TI - Genetics of quantitative variation in human gene expression. PMID- 15338643 TI - Regulation of alpha-synuclein expression: implications for Parkinson's disease. PMID- 15338644 TI - Genomic perspective and cancer. PMID- 15338645 TI - Chromosome 21 and Down syndrome: the post-sequence era. PMID- 15338646 TI - Harvesting the genome's bounty: integrative genomics. PMID- 15338647 TI - Genomic disorders: genome architecture results in susceptibility to DNA rearrangements causing common human traits. PMID- 15338648 TI - Human versus chimpanzee chromosome-wide sequence comparison and its evolutionary implication. PMID- 15338649 TI - Novel transcriptional units and unconventional gene pairs in the human genome: toward a sequence-level basis for primate-specific phenotypes? PMID- 15338650 TI - Positive selection in the human genome inferred from human-chimp-mouse orthologous gene alignments. PMID- 15338651 TI - mtDNA variation, climatic adaptation, degenerative diseases, and longevity. PMID- 15338652 TI - Inferring human history: clues from Y-chromosome haplotypes. PMID- 15338653 TI - The new quantitative biology. PMID- 15338654 TI - Study finds large cost variation for laparoscopic gastric bypass. PMID- 15338655 TI - What it takes to meet the goal of being nation's 'safest hospital'. PMID- 15338656 TI - Five keys to effective surgical services. PMID- 15338657 TI - What new Stark rules mean for ORs. PMID- 15338658 TI - Rewards and recognition for CS techs. PMID- 15338659 TI - Getting to the bottom of case cart errors. PMID- 15338660 TI - Are you up to speed on HIPAA security? PMID- 15338661 TI - Some liposuctions exceed guidelines. PMID- 15338662 TI - Multiple primary malignant neoplasms in urologic patients. AB - METHODS: Second primary cancers constitute approximately 9-10% of malignancies diagnosed in the United States. We aimed to show the risk and incidance of second primary tumor occuring associated to urologic tumors and show the distrubution of tumors in Tracia region. We retrospectively examined the patients' files with the diagnosis of malignant disease between the years 1985-2000. Hazard function analysis was performed to estimate the relative risk of secondary malignancy occuring. Age, sex and tumor number were examined to find out if they affect on mortality rate. RESULTS: A total number of 25 MPMNs were diagnosed. In 52 percent of the cases the second primary neoplasm developed within six months. The relative risk of development of a second neoplasm is found as increasing 1.111 times per month. The incidence of secondary malignancy occuring in the patients with one tumor was found as 6.31%. Age (p < 0.001) and tumor number (p < 0.001) are found as statistically effective predictor on mortality rate where the sex is not. CONCLUSIONS: In the patients with a primary tumor not only the metastasis must be investigated but also second primary tumors should be taken into consideration. PMID- 15338663 TI - Laparoscopic management of symptomatic simple renal cysts. AB - OBJECTIVE: To evaluate the efficacy of retroperitoneoscopy for the treatment of symptomatic renal cyst disease. PATIENTS AND METHODS: We evaluated the efficacy and morbidity of aspiration versus open and laparoscopic surgical techniques in a paired institution over a 20-year period. Prospectively recorded parameters for the 11 cases dealt with by primary retroperitoneoscopic techniques were compared with historical controls dealt with by open surgery. RESULTS: Small volume cysts were satisfactorily dealt with by aspiration alone. All patients undergoing laparoscopic cyst decortication for the larger volume cysts had a satisfactory outcome with improvement in their symptoms and objective cure on follow up imaging studies. CONCLUSION: Laparoscopic management for all cysts is a safe, effective and minimally invasive alternative to open surgery for symptomatic renal cysts. Simple drainage under radiological guidance is also highly effective and should probably be first line treatment for cysts up to 6 cms in diameter whereas primary laparoscopic treatment should be suggested for larger cysts. PMID- 15338664 TI - Bilateral collecting duct carcinoma presenting with tumour associated nephritis and end-stage renal failure. A case report and review of the literature. AB - We present a 55 year old male who was investigated for painless macroscopic haematuria and had essentially normal radiological and cystoscopic findings. He progressed rapidly and was eventually diagnosed with a Collecting Duct Carcinoma. This case is of interest as it is the first reported case of Collecting Duct Carcinoma occurring bilaterally. It is also the first case to cause end stage renal failure requiring dialysis due to extensive tubular involvement. Finally, it is the first time this malignancy has been found to cause a tumour associated nephritis. We describe the clinical course and present the various histological findings followed by a review of the literature. PMID- 15338665 TI - Spontaneous perirenal hemorrhage: a 10-year experience at our institution. AB - OBJECTIVE: To report our experience in patients with spontaneous perirenal hemorrhage (SPH) seen at our institution over a 10-year period. MATERIAL AND PATIENTS: Over the years from 1992 to 2002, 13 patients with SPH without a history of trauma, were treated at our hospital. There were 5 male and 8 female patients with a mean age of 55.7 years (range 36-79 years). The patients' records were reviewed retrospectively with respect to etiology, clinical presentation, radiologic findings and therapeutic management of SPH. RESULTS: All patients were presented with flank or abdominal pain. Radiological evaluation included ultrasonography (U/S) in 7 cases and computed tomography (CT) in 13 cases. An underlying renal mass was indentified employing U/S in 2 cases and using CT in 10 cases respectively. The etiology of SPH was determined in 12 cases. The most common causes were angiomyolipoma (5 patients) and renal cell carcinoma (4 patients). Out of the remaining 4 cases with SPH, one was associated with anticoagulant therapy; polyarteritis nodosa and Wegener angeitis were the underlying diseases in 2 cases respectively; finally, the etiology could not be determined in 1 case. All but two patients were managed surgically. Complete nephrectomy was performed in 6 cases, partial nephrectomy in 4 and simple evacuation of the haematoma was performed in 1 case. CONCLUSIONS: SPH presence should arouse suspicions concerning its etiology, since the most common cause is a renal tumor and approximately 50% of such tumors are malignant. CT scanning is a useful imaging modality for the initial evaluation of SPH, permitting identification of the underlying cause in most instances. PMID- 15338666 TI - Case report: Minimally invasive treatment of an unusual obstructive ectopic upper pole ureter. AB - An 18-year old man presented with left loin pain and recurrent urinary tract infections. Ultrasound and IVU revealed a duplex left kidney with hydronephrosis of the upper pole moiety. F-15 Mag-3 renography confirmed obstruction of the upper pole moiety. Cysto-urethroscopy identified the site of ectopic insertion to be adjacent to the verumontanum in the prostatic fossa. Retrograde ureterography showed the level of obstruction and dilation of the stricture was carried out. Post operatively, the patient became asymptomatic and remains so at 5-year follow up. This is an unusual case of obstructive upper pole ureter with an ectopic insertion into the prostatic fossa. We also demonstrated that dilatation of a stricture at this site can achieve good long term outcome. PMID- 15338667 TI - Recurrent flank pain caused by eosinophilic ureteritis mimicking urinary stone disease: a case report. AB - Flank pain is caused by a variety of pathologies of which urinary stone disease is the most frequent. Eosinophilic ureteritis is a rare stenosing condition of the ureter. Eosinophilic ureteritis can cause flank pain and/or unilateral hydronephrosis. On pathological examination it is characterised by a marked infiltration of the submucosal layers by eosinophils. A relationship of this condition with atopy, hypereosinophilic syndrome and prior ureteral trauma has been described. Surgical resection of the stenosing segment with end-to-end anastomosis is usually a successful treatment. In some cases of proximal disease total nephro-ureterectomy has been performed. One author describes remission of disease after a prolonged oral corticosteroid regimen. In this article we report another case of eosinophilic ureteritis and discuss different treatment strategies. PMID- 15338668 TI - The secret of the phantom stone: a case report. AB - A 50-year old man, who has a history of allergies to i.v. contrast and NSAID, was presented with right renal colic. He was treated with regular opiate injections. Plain abdominal X ray showed a 1 cm opacity at the level of L2 but spiral CT failed to locate the urinary calculus. Abdominal examination revealed a small piece of metal implanted in the subcutaneous anterior abdominal wall which was also subsequently identified on CT scanning. When the opiate treatment was withdrawn, the patient absconded. PMID- 15338669 TI - Vesical pacing in patients with overactive bladder: technique and results. AB - OBJECTIVES: A recent study has demonstrated that the electric activity of the overactive bladder (OAB) is 'dysrhythmic'. The cause was attributed to a disordered vesical pacemaker which discharges these waves. In a subsequent study, the dysrhythmic waves have been 'normalized' by vesical pacing and the optimal parameters which are required to achieve normalization have been defined. We investigated the hypothesis that vesical pacing of the OAB might improve not only the vesical electric activity but also the symptoms. METHODS: Vesical pacing was used in 9 patients (age 39.2 +/- 10.3; 5 women, 4 men) with OAB. Under anesthesia, the pacemaker was implanted in an inguinal subcutaneous pocket and connected to 2 pacing electrodes implanted into the vesical vault. The normalization of the waves was tested by 2 recording electrodes which were temporarily applied to the vesical wall and removed post-testing. The pacemaker was then programmed for home pacing to be activated at given times. RESULTS: Vesical pacing effected normalization of the dysrhythmic electric waves with disappearance of the OAB symptoms in 7 patients and failed in 2. Vesical pacing was abandoned in 3/7 patients after a few months following the spontaneous disappearance of the symptoms. CONCLUSIONS: Vesical pacing has normalized the dysrhythmic electric activity and suppressed the symptoms of the OAB in 77.7% of patients. The pacemaker was removed in 5 patients: 2 failures and 3 after spontaneous waves normalization. No complications were encountered. Vesical pacing is suggested as a treatment for OAB when commonly used therapeutic modalities have failed. PMID- 15338670 TI - The use of human dura mater as an interposition graft in the treatment of vesicovaginal fistula. AB - PURPOSE: In this prospective study we aimed to describe our new technique; dehydrated human dura mater as an interposition graft in patients undergoing benign vesicovaginal fistula repair (VVF). PATIENTS AND METHODS: Eleven patients with benign VVFs aged 38-73 years were operated with dura mater interposition technique at our institution between July 1996 and February 2002. In all patients fistula were secondary to abdominal hysterectomies with benign diseases and two had undergone previous transvaginal surgery. Patients were operated 8-15 (mean 12) weeks after previous surgery. A suprapubic cystotomy was performed and the fistula was not excised. The bladder wall is widely dissected from underlying vagina. Dura-mater was placed over the vaginal suture line to close the fistulous tract, cytostomy was inserted and the bladder was closed. An 18 F Foley catheter was left in the bladder for 5 days. A successful repair is defined as no leakage by cystogram at 14 days postoperatively and completely dry by patient report. RESULTS: An overall success rate of 100% was achieved in all patients. All patients remain dry at the follow-up which ranges from 7 to 60 (mean 26) mounts. Patients were discharged at 14 to 26 (mean 16) days postoperatively. No major complications or side effects were observed during the follow-up period. CONCLUSIONS: The use of human dura mater as an interposition graft in the treatment of vesicovaginal fistula is an alternative for VVF repairs because of its excellent tissue compability, stability, good elasticity and absorbability. PMID- 15338671 TI - T1G3 bladder cancer--indications for early cystectomy. AB - OBJECTIVES: To review our experience with early radical cystectomy in patients with T1G3 Transitional Cell Carcinoma of bladder (TCC). PATIENTS AND METHODS: Thirty patients, who underwent early radical cystectomy over a 10-year period for clinical stage T1G3 TCC bladder, were studied. Of these 21 (70%) had radical cystectomy without treatment with intravesical chemo/immunotherapy. The number of tumours, presence or absence of Carcinoma In-Situ (CIS) and the pathological stage of the cystectomy specimen were recorded in each patient. Disease specific survival was determined in the subgroups using Kaplan-Meier estimates. RESULTS: Seventeen patients underwent radical surgery for a single tumour without concomitant CIS (Group A). The other 13 had multiple tumours with or without concomitant CIS or a single tumour with CIS (Group B). The disease was upstaged after cystectomy in 1 (6%) patient in Group A compared to 7 (55%) in Group B, (p = 0.009). Nine (53%) had pT0 disease in Group A compared to 0% in Group B, (p = 0.0017). The 5-year cancer specific survival rates were 92% in Group A and 82% in Group B. CONCLUSIONS: In patients with multiple T1G3 tumours with or without associated CIS, or in those with single T1G3 tumour with associated CIS the incidence of the disease being already muscle invasive at the time of clinical diagnosis is 55%. Early radical cystectomy should be advocated in this group. Conversely, for a single T1G3 tumour without associated CIS, conservative bladder preserving strategy with immuno-chemotherapy and close surveillance is justified. PMID- 15338672 TI - Chondrosarcoma of the pubis imitating bladder tumor radiologically: a case report. AB - Chondrosarcoma is a rare tumor that more frequently involves the pelvic bones, the femur, and the humerus. The rareness of the tumor makes the diagnosis difficult. Malignant cartilage tumors typically have an unstable radiographic appearance. In this report we aimed to point out the difficulties of the radiological differentiation of the pelvic chondrosarcoma and bladder tumor. PMID- 15338673 TI - Are indications for prostatectomy in octogenarians the same as for younger men? AB - Prostatectomy for benign hypertrophy of the prostate is usually performed to alleviate lower urinary tract symptoms (LUTS). We assessed indications for and risks of prostatectomy in men 80 years of age and compared them to those for younger men in order to determine whether indications for prostatectomy in octogenarians are different than these for younger men. Medical records of 171 men comprised of 84 patients >80 years of age (mean 84.4) and 87 patients <65 years of age (mean 60.6) who underwent prostatectomy for benign prostate hypertrophy were reviewed. Data regarding indications for surgery, American Society of Anesthesiologists system grade, anesthesia and surgery performed, duration of hospitalization and intrahospital postoperative complications were obtained. The respective indications for surgery in the very elderly and younger patients were: urinary retention with indwelling catheter in 46 (55%) and 34 (39%) (p < 0.04), LUTS in 32 (38%) and 52 (59%) (p < 0.005), and gross hematuria in 6 (7%) and 1 (1.2%). Transurethral prostatectomy was performed in 47 elderly patients (56%) and in 30 young patients (34.5%). The other patients in each group underwent open (suprapubic prostatectomy) surgery. The overall complication rate was significantly higher in the elderly group (39% vs 22%, p < 0.05), with major complications occurring only in this group. Indications for surgery were different for octogenarians than for younger men. Morbidity and mortality rates were significantly higher among the elderly men. Age appears to be an independent risk factor for complications associated with prostatectomy. PMID- 15338674 TI - Adenoid-basal cell tumor of the prostate gland. A case report: histomorphologic and immunohistochemical features. AB - Lesions with basaloid features are very rare and controversial within the prostate gland. There seems to be a morphologic continuum ranging from focal basal cell hyperplasia (BCH) to florid adenoid basal cell tumor (ABCT). The clinicopathological features of a basaloid lesion which was associated with an acinar prostatic carcinoma of the prostate gland are described in a 74-year-old man, and discussed under the light of the literature. PMID- 15338675 TI - A case report: Leukaemic infiltration and hyperplasia of the prostate. AB - Leukaemic infiltration of prostate with hyperplasia is a rare manifestation and is usually found in known cases of leukaemia, but it may be the first sign of an undiagnosed leukaemia. In this report, a rare case of leukaemic infiltration of the prostate in a 64-year-old man is represented. PMID- 15338676 TI - The effect of prostate cancer and antiandrogenic therapy on lipid peroxidation and antioxidant systems. AB - In living organism, excessive free radicals or oxidative damage which occur as a result of deficient antioxidant defensive mechanisms by the effect of endogenous and exogenous factors, influences especially developmental steps of chemically induced cancers. In our study, plasma malondialdehyde level (MDA) as an indicator of lipid peroxidation, erythrocyte glutathione (GSH) level as an indicator of antioxidant state, glutathione reductase (GSH-Red), glutathione peroxidase (GSH Px), glutathione-S-transferase (GST) as an antioxidant enzymes and plasma vitamin E level were detected in patients with prostate cancer (21 males; age, 69.4 +/- 4.8 years) before and after three months of antiandrogenic therapy with goserelin acetate as luteinizing hormone releasing hormone (LHRH) analogue. Healthy people evaluated as a control group (20 males; age, 63.7 +/- 3.9). Erythrocyte GSH levels, the activities of GSH-Red and GSH-Px and plasma vitamin E levels were found significantly low in patients with prostate cancer when compared with the healthy subjects (p < 0.01, p < 0.05, p < or = 0.001 and p < or = 0.001 respectively). Plasma MDA level and erythrocyte GST activity of patient group were significantly higher than the levels of control group (p < or = 0.001 and p < or = 0.001 respectively). After antiandrogenic therapy erythrocyte GSH level, GSH-Red, GSH-Px activity and plasma vitamin E level were found unchanged. Significant decrease in plasma MDA level and significant increase in erythrocyte GST activity were detected in patient group (p < 0.05 and p < or = 0.01 respectively). The study has revealed the shift in the oxidant-antioxidant balance towards oxidative state in patients with metastatic prostate cancer. Our results showed that antiandrogenic therapy increased in GST activity, decreased in lipid peroxidation. PMID- 15338677 TI - A case report: Prostatic carcinoma with metastasis to the testicle. AB - Secondary testicular tumours are rare. We present a 54 years old patient with testicular metastasis from carcinoma of the prostate with a 7 years history of hormonotherapy and chemotherapy for advanced carcinoma. PMID- 15338678 TI - Case report: Nonobstructive giant urethral stone with two safety pins. AB - The urethra can be the site of various types of foreign bodies. A 61-year-old man having a giant urethral stone with two safety pins was evaluated according to the literature. To our knowledge, we report the first case of foreign bodies in urethra in which existing non-obstructive giant urethral stone with two safety pins. PMID- 15338679 TI - The potential role of inducible nitric oxide synthase (iNOS) activity in the testicular dysfunction associated with varicocele: an experimental study. AB - Nitric oxide (NO) has been reported to be increased in the spermatic veins of men affected by varicocele. The aim of the present study was to determine whether iNOS (inducible nitric oxide synthase) has a role in testicular dysfunction associated with varicocele, immunohistochemistry analyze was used to study iNOS activity in testis of adolescent rats with experimental left varicoceles. Rats were randomly divided into three groups. The first group consisted of rats undergoing partial ligation of left renal vein (n:12). The second group consisted of rats undergoing a sham operation (n:6) and, the third group referred to as control rats (n:7). Immunohistochemistry slides were evaluated by counting the number of positive cells and expressed as percents (% iNOS activity). We found that iNOS was predominantly expressed in the cytoplasm of Leydig cells in each group and only a small amount of iNOS was expressed in Sertoli cells. There were significant differences in % iNOS activity between both testes of varicocele group and both of testes control group (p < 0.01), but no significant differences were noted between other groups (p > 0.05). Because of iNOS activity was markedly increased in the Leydig cells of varicocele bearing rats, we suggest that iNOS activity may play a role in the testicular dysfunction associated with varicocele during adolescence. PMID- 15338680 TI - A case of testicular rhabdomyosarcoma. AB - Pure testicular rhabdomyosarcoma is a very rare tumor and 7 cases have been reported in literature. A 20-year-old male patient presented with a painless right testicular swelling who underwent inguinal orchiectomy with suspicion of testicular malignancy. The case was regarded as an embryonal rhabdomyosarcoma according to histopathologic and immunohistochemical findings. The case is presented and the relevant literature is reviewed and discussed. PMID- 15338681 TI - Primary adenocarcinoma of the epididymis: case report. AB - Primary epididymal malignancies are uncommon and usually benign. Benign paratesticular tumors are most commonly adenomatoid, while the most common malignant paratesticular tumors are rhabdomyosarcomas. Approximately 25% of all epididymal tumors are malignant, and of the benign tumors, 60% to 78% are adenomatoid tumors. According to a recent MEDLINE search using epididymis and adenocarcinoma as key words, reports of a primary epididymal adenocarcinoma are extremely rare with only 23 cases in the literature. We report a case of epididymal adenocarcinoma with clinical follow up and metastatic natural history of this rare malignancy. PMID- 15338682 TI - Expression of alpha5beta1 and alpha6beta1 integrins in IgA nephropathy (IgAN) with mild and severe proteinuria. An immunohistochemical study. AB - Immunoperoxidase staining was carried out using monoclonal antibodies against integrin alpha5beta1 and integrin alpha6beta1 on renal biopsy specimens from patients with IgA nephropathy with mild proteinuria (n = 15) and with severe proteinuria (n = 10). Our study revealed increase in glomerular immunoexpression of alpha5beta1 in renal biopsies in IgA nephropathy with severe proteinuria. There were no statistical differences between interstitial alpha5beta1 immunostaining, as well as tubular alpha6beta1 immunoexpression in renal tissue between patients with mild and severe proteinuria. The intensity of interstitial alpha5beta1 integrin immunoexpression was positively correlated with the degree of interstitial fibrosis in both studied groups, meanwhile the intensity of tubular alpha6beta1 integrin immunoexpression was not related to the degree of renal interstitial fibrosis in patients with mild and severe proteinuria. Our results suggest that elevated immunoexpression of alpha5beta1 integrin on endothelial glomerular cells in the renal tissue in patients with severe proteinuria may point to the role of this integrin in the mechanism of glomerular injury in these cases of IgA nephropathy. The positive association between the interstitial expression of alpha5beta1 integrin and the relative interstitial cortical volume in renal biopsies in patients with mild and severe proteinuria suggests that alpha5beta1 integrin may play a role in the pathogenesis of chronic progressive renal disease in both studied group. Lack of positive correlation between tubular alpha6beta1 integrin immunoexpression and the relative interstitial cortical volumen may indicate that this molecule play no role in the pathomechanism of interstitial fibrosis in IgA nephropathy with mild and severe proteinuria. PMID- 15338683 TI - Prevalence of malnutrition in Nigerians with chronic renal failure. AB - In developed countries, malnutrition is common in patients with chronic renal failure (CRF) and has adverse effects on patient morbidity and mortality. The prevalence of malnutrition before the initiation of dialysis is poorly characterized in CRF patients in developing countries. We studied the prevalence of malnutrition among Nigerians with CRF before commencement of dialysis. Body mass index (BMI) and serum protein levels were measured in 74 dialysis naive Nigerians with CRF and 48 controls. Patients with nephrotic syndrome, steroid use and failure of organs other than the kidneys were excluded. The mean BMI was significantly lower in the patients compared to the controls (22.4 +/- 14.9 kg/M2 Vs. 25.2 +/- 2.7 kg/M2; p = 0.0001). Low BMI (less than 20 Kg/M2) was present in 16 (21.6%) of the patients compared with one of the controls. The mean serum total protein and albumin were also significantly lower in the patients compared to controls (61.9 +/- 14.4 g/L Vs. 73.8 +/- 6.8 g/L; p < 0.0001, and 31.5 +/- 9.3 g/L Vs. 39.6 +/- 4.4 g/L; p < 0.0001 respectively). Protein malnutrition (serum albumin < 29 g/L) was present in 32 (43.2%) of patients with CRF and one (2.1%) of the control subjects. Malnutrition is common in Nigerian CRF patients before the commencement of dialysis. In these patients, emphasis should be placed on prevention and/or correction of malnutrition because of its documented adverse effects on the outcomes of maintenance dialysis. PMID- 15338684 TI - Bone mineral density in live related kidney transplant children and adolescents. AB - Successful kidney transplantation corrects many of the metabolic abnormalities associated with development of renal osteodystrophy, but despite a well functioning graft, osteopenia, remains prevalent in adult and pediatric kidney recipients. The factors that affect the bone mineral density (BMD) and the long term course of BMD after transplantation in children is still unknown. We performed a cross sectional study to determine BMD in 83 recipients who received living renal allotransplants in Mansoura Urology & Nephrology Center between 1981 and 2002 (mean age at transplantation 13.2 +/- 3.1 years) by dual energy x-ray absorptiometry at various time intervals up to 16 years after transplantation (mean duration after transplantation was 48 +/- 34 months, range 12-192 months). The mean +/- SD for BMD was -2.28 +/- 2.06 for lumbar 2-4 spine and -1.44 +/- 1.44 for the total body BMD as corrected for body surface area. Osteopenia/osteoporosis were present in about two thirds of our kidney transplant recipients. The significant risk factors for osteopenia/osteoprosis using univariate analysis were the cyclosporine based immunosuppressive regimen, cumulative dose of steroids/m2 surface area, graft dysfunction and the urinary deoxypyridinoline. Using logistic regression analysis the cumulative steroid dose/m2 surface area and the urinary deoxypyridinoline were the major significant predictors for bone loss. In conclusion, osteopenia and osteoprosis are common in pediatric and adolescent renal transplant patients. The cumulative steroid dose and the urinary deoxypyridinoline were the major predictors for bone loss. PMID- 15338685 TI - Sodium and volume overload in peritoneal dialysis: limitations of current treatment and possible solutions. AB - Cardiovascular disease is a leading cause of death in patients with chronic kidney disease. Recent evidence suggests that hypertension and subclinical volume expansion is common in patients on peritoneal dialysis. Moreover, recent studies pointed out that sodium removal is limited in patients on peritoneal dialysis and mortality has been shown to co-relate with fluid and sodium removal. Treatment of sodium and fluid removal includes dietary salt and fluid restriction, use of diuretics, icodextrin, strategies also considered helpful to control hypertension. Despite availability of these measures, prevalence of hypertension remains high in PD patients. Hence, innovative strategies are urgently required to address this common and difficult clinical problem. This article reviews limitations of available measures to manage sodium and fluid overload and hypertension and suggests possible role and place of low sodium dialysis solutions in PD patients. PMID- 15338686 TI - Reproducibility of the measurements of creatinine clearance in patients with a stable renal function. AB - In everyday practice the measurement of creatinine clearance (CCr) is used for the evaluation of the renal function. Since large body of evidence points to the inaccuracy of this test, we decided to check whether a repeated measurement of CCr might improve the assessment of the glomerular filtration rate. The study comprised 233 subjects, 105 females and 128 males, aged 22-80 years (mean age: 50.8 +/- 12.8 years) with the history of renal stones and a stable renal function, e.g. without changes in plasma creatinine. In every patient CCr was measured twice. Thereafter, all studied subjects were sorted by the increasing absolute values of the difference between two measurements of CCr e.g. as the difference between two measurements of CCr with omitted sign (absDCCr). Then, they were divided into 3 groups corresponding to the tertiles of absDCCr (tertile = 1/3). When the two values of CCr were compared among all patients, the mean difference between them (DCCr) was only 1.51 +/- 26.86 ml/min. The values of the individual DCCr varied, however, from -122.2. ml/min to 69.3 ml/min. As far as sex, age and height were concerned, there were no differences between the groups. However, weight was lower in group A (first tertile) vs. group B (second tertile) (p < 0.05). Also, BMI was lower in the group A vs. both group B and C (p < 0.05 and p < 0.01; respectively). In conclusion, our study points out to a poor reproducibility of the measurements of CCr in non-hospitalized subjects, especially in those with the high BMI. PMID- 15338687 TI - Beneficial role of intravenous calcitriol on bone mineral density in children with severe secondary hyperparathyroidism. AB - OBJECTIVES: In this prospective study, the effect of calcitriol therapy on bone mineral density and osteopenia in patients with severe secondary hyperparathyroidism has been investigated. MATERIALS AND METHODS: The study was carried out on 24 chronic dialysis patients consisting of 13 boys and 11 girls, aged between 8-18 years. Patients were divided into 3 groups according to the severity of hyperparathyroidism and therapy regimens. Group I consisted of 5 patients with normal parathormon levels who did not receive calcitriol therapy. In group II and III, there were patients with secondary hyperparathyroidism. Group II consisted of 10 patients receiving oral calcitriol therapy. Group III consisted of 9 patients receiving intravenous (i.v.) calcitriol. Bone mineral density was measured by dual energy x-ray absorptiometry. Osteopenia was defined as a Z-score worse than -2. Bone mineral density was assessed as baseline and at the end of one year. RESULTS: A significant improvement was observed in Z-score in the group III whereas the mean value of Z-score tended to be worse in group I and it was not significantly different in group II from the initial values. The better Z-score in group III was associated with more effective stabilization of alkaline phosphatase level and bone specific alkaline phosphatases (BAP) concentrations. CONCLUSION: Significant improvement of Z-score in group III suggested the beneficial role in i.v. administration of calcitriol in chronic dialysis patients. PMID- 15338688 TI - Nephropathology quiz page. Heterozygous Fabry's disease. PMID- 15338689 TI - Membrane bioreactor technology and sustainable water. PMID- 15338690 TI - Effects of nitrosation on the formation of cyanide in publicly owned treatment works secondary effluent. AB - Cyanide has been detected in the effluents of some publicly owned treatment works (POTWs) at levels exceeding the influent concentration. The presence of nitrite ion (NO2-) as a common constituent in domestic wastewater effluents may play an important role in the formation of cyanide through reaction with certain kinds of organic compounds, especially aromatic compounds. Laboratory studies with seven organic compounds (aniline. p-toluidine, phenol, 1,2,4-trihydroxybenzene, L serine, glycine, and benzoic acid) revealed that cyanide can be formed by reaction of nitrite with some of these compounds. The most substantial free cyanide (HCN. CN-) production observed at 25 degrees C was 0.15 mg/L from reaction of 0.01 mM 1.2.4-trihydroxybenze with 5 mg/L nitrite for 72 hours. Substantial free cyanide formation was also observed at pH 2-4 in experiments with POTW effluents when reactive organics and nitrite were both added to wastewater. Formation of cyanide through nitrosation was strongly pH dependent, being most significant at low pH (2 to 4) and negligible at neutral-to-high pH. This result points to nitrous acid (HNO2) as being more reactive than the dissociated NO2- ion. The reaction of these nitrite species with organics also occurs in conventional analyses for total cyanide which involve distillation under strongly acidic conditions. Sufficient sample pretreatment with sulfamic acid at the time of sampling, not at the time of analysis. is highly recommended to prevent biasing analytical measurement of total cyanide in POTW effluents. PMID- 15338691 TI - Effects of thiocyanate on the formation of free cyanide during chlorination and ultraviolet disinfection of publicly owned treatment works secondary effluent. AB - Cyanide has been detected in effluent of some publicly owned treatment works (POTWs) at levels exceeding influent concentration. Thiocyanate (SCN-) is a cyanide-related compound encountered in most POTW influents and may be decomposed to free cyanide (CN-) under some circumstances. Effects of SCN- on the formation of cyanide during chlorination and UV disinfection were studied through a laboratory study with synthetic solutions and POTW secondary effluent. Results indicated that CN- was detected in SCN- solutions after chlorination in which the chlorine dose or reaction time was not sufficient to destroy SCN-completely, thus ensuring no residual chlorine to destroy any CN-produced. It was also found that SCN can be broken down to yield cyanide by UV irradiation. In addition, SCN- was observed to cause significant positive error in the conventional total cyanide analysis technique involving acidic distillation and colorimetric measurement when nitrate was present in the sample. PMID- 15338692 TI - Activated sludge deflocculation in response to chlorine addition: the potassium connection. AB - Chlorination is often used to control filamentous bulking in activated sludge systems. Pure culture and mixed-liquor experiments showed that soluble potassium (K+) concentrations increased by 2.4 mg/L (80%) and 1.5 to 3.6 mg/L (11 to 30%) in the bulk liquid phase of pure and activated sludge cultures that were exposed to chlorine, relative to unchlorinated controls. Effluent turbidity and total suspended solids from settled mixed liquor increased significantly in both short term batch and sequencing batch reactor experiments when chlorine mass load increased above 6 milligrams of chlorine per gram mixed liquor volatile suspended solids (mg Cl2/g MLVSS) in a single dose, which correlated with a localized chlorine concentration at the dose point of 10 mg/L as Cl2 or greater. The results support the hypothesis that the glutathione-gated potassium efflux (GGKE) bacterial stress response may contribute to increased effluent turbidity associated with high doses of mixed-liquor chlorination. It is suggested that potassium is a useful parameter to monitor at full-scale facilities when determining chlorine mass doses that should be used to control filaments and minimize increases in effluent turbidity. PMID- 15338693 TI - Use of naturally growing aquatic plants for wastewater purification. AB - This paper examines potential uses of naturally growing aquatic plants for wastewater purification. These plants enhance the removal of pollutants by consuming part of them in the form of plant nutrients. This applies to urban and agricultural wastewater, in particular, where treatment units of different sizes can be applied at the pollution source. The effectiveness of wastewater purification by different plants was tested on laboratory and pilot scales. The growth rate of the plants was related to the wastewater content in the water. Batch and semicontinuous experiments verified that the plants are capable of decreasing all tested indicators for water quality to levels that permit the use of the purified water for irrigation. This applies to biochemical oxygen demand (BOD), chemical oxygen demand, total suspended solids. pH, and turbidity. In specific cases, the turbidity reached the level of drinking water. Comparison of BOD concentrations with typical levels in water treatment facilities across the country indicates the effectiveness of water purification with plants. A major effect of treatment with plants was elimination of the disturbing smell from the wastewater. It is shown that mixtures of wastewater and polluted water from the Kishon River are amenable in varying degrees to treatment by the plants. The higher the wastewater content in the mixture, the more effective the treatment by the plants. In this context, a scheme for rehabilitation and restoration of the Kishon River is presented and technical and economical aspects of the purification technology are considered. PMID- 15338694 TI - An experimental study of air entrainment and oxygen transfer at a water jet from a nozzle with air holes. AB - An adequate supply of dissolved oxygen is important in natural rivers and in some water treatment processes. The dissolved oxygen concentration can be enhanced by entraining air bubbles in a receiving pool. When a water jet impinges a receiving pool at rest, air bubbles may be entrained and carried away below the pool free surface. This process is called plunging water jet entrainment and aeration. This paper describes an experimental study of the air entrainment rate and oxygen transfer efficiency of circular nozzles with and without air holes. In particular, the effect of varying the number, positions, and open/close status of the air holes is investigated. A negative pressure occurred depending on the air holes opened on the circular nozzles. This phenomenon affected the water jet expansion, water jet shape, air entrainment, and bubble penetration depth and, hence, the oxygen transfer efficiency. It was demonstrated that the air entrainment rate and the oxygen transfer efficiency of the circular nozzles with air holes were better than those of the circular nozzles without air holes. Therefore, adding air holes to a simple, circular nozzle could lead to a significantly increased air entrainment rate and oxygen transfer efficiency. PMID- 15338695 TI - Analyzing the uncorrected error of dilution water demand for the dilution biochemical oxygen demand method. AB - Dilution water demand (DWD) can cause a positive error when the dilution biochemical oxygen demand (BOD) method is used. Dilution water demand may be attributed to oxidation of organic impurities in the dilution water and nitrification of ammonia added as a nutrient. To minimize the error associated with these sources, the standard BOD method requires that DWD be less than 0.2 mg/L in 5 days and does not allow correction for DWD when calculating test results. This study derives a set of theoretical equations to analyze the uncorrected errors with and without seeding. The authors concluded that DWD can be completely corrected if seeded dilution water is used for the sample dilution. When seeding individual bottles, the uncorrected error approaches 8.3 to approximately 8.8% at a 5-day depletion of 2 mg/L for a typical secondary effluent. Tests without seeding show an almost 1% higher uncorrected error than seeded tests. The analysis also suggests that these errors can be effectively reduced to less than 3% when the 5-day depletion approaches 6 mg/L. even for 5 day biochemical oxygen demand concentrations exceeding I x 10(4) mg/L. Further analysis indicates that, if not inhibited, the ammonium added to dilution water as a nutrient may contribute additional error due to nitrification. PMID- 15338696 TI - Comparison of E. coli, enterococci, and fecal coliform as indicators for brackish water quality assessment. AB - Escherichia coli (E. coli), enterococci, and fecal coliform data were collected and compared as potential indicators for swimmablility assessment of a brackish waterbody (Lake Pontchartrain, Louisiana). These indicators were measured during lake background conditions, in stormwater runoff (before dilution with lake water), and in the outfall plume within the lake following storm events. Microbial indicator titers associated with suspended particles and lake-bottom sediments were also investigated. Overall reduction rate constants for fecal coliform, E. coli, and enterococci in lake water and sediment were measured and reported. Attachment of microbial indicators to suspended matter and subsequent sedimentation appeared to be a significant fate mechanism. A slower reduction rate of indicator organisms in sediment further suggested that bottom sediment may act as a reservoir for prolonging indicator organism survival and added concern of recontamination of overlaying waters due to potential solids resuspension. Results indicated that enterococci might be a more stable indicator than E. coli and fecal coliform and, consequently, a more conservative indicator under brackish water conditions. PMID- 15338697 TI - Relative efficacy of intrinsic and extant parameters for modeling biodegradation of synthetic organic compounds in activated sludge: dynamic systems. AB - The utility of intrinsic and extant kinetic parameters for simulating the dynamic behavior of a biotreatment system coupled with a distributed, unstructured, balanced microbial growth model were evaluated against the observed response of test reactors to transient loads of synthetic organic compounds (SOCs). Biomass from a completely mixed activated-sludge (CMAS) system was tested in fed-batch reactors, while a sequencing batch reactor (SBR) was tested by measuring SOC concentrations during the fill and react period. Both the CMAS system and the SBR were acclimated to a feed containing biogenic substrates and several SOCs, and the transient loading tests were conducted with biogenic substrates along with one or more SOCs. Extant parameters more closely reflect the steady-state degradative capacity of activated-sludge biomass than intrinsic parameters and, hence, were expected to be better predictors of system performance. However, neither extant nor intrinsic parameters accurately predicted system response and neither parameter set was consistently superior to the other. Factors that may have contributed to the inability of the model to predict system response were identified and discussed. These factors included the role of abiotic processes in SOC removal, disparity in the bases used to evaluate parameter estimates (substrate mineralization) and reactor performance (substrate disappearance), inhibitory substrate interactions under the severe loading conditions of the SBR, changes in the physiological state of the biomass during the transient loading tests, and the presumed correlation between the competent biomass concentration and the influent SOC concentration. PMID- 15338698 TI - Can chemical oxidation improve the permeability of infiltration basins? AB - The Rapid Infiltration and Extraction (RIX) facility, a soil aquifer treatment system, began taking secondary effluent from the City of San Bernardino, California, in 1996. The gradual decrease in the hydraulic conductivity of the infiltration basins at RIX has been attributed to the accumulation of organic matter in the surface sand. Periodic tillage of the surface sand to restore the permeability has mixed this organic matter to a depth of nearly 50 cm. We hypothesized that in situ chemical oxidation of the surface sand might improve the infiltration rate and increase the time between filling and drying cycles. The effect of organic matter oxidation on sand permeability was tested in laboratory sand columns treated with sodium hypochlorite, calcium hypochorite, and ozone gas. All oxidants significantly decreased the hydraulic conductivity of the surface sand. The loss in permeability was attributed to an increase in dispersed clay plus silt-sized particles that were released as a result of oxidation. This study suggests that ex situ sand-washing operations, currently being used to clean the sand, could be improved by the addition of oxidants to the wash water. PMID- 15338699 TI - Behaviors of intercellular materials and nutrients in biological nutrient removal process supplied with domestic wastewater and food waste. AB - A four-stage biological nutrient removal (BNR) process was operated to investigate the effect of anaerobically fermented leachate of food waste (AFLFW) as an external carbon source on nutrient removal from domestic wastewater having a low carbon-to-nitrogen ratio. The BNR system that was supplemented with AFLFW showed a good performance at a sludge retention time (SRT) of 30 days, despite low temperature. With this wastewater, average removal efficiencies of soluble chemical oxygen demand (COD), total nitrogen (T-N), and total phosphorus (T-P) were 88 to 93%, 70 to 74%, and 63 to 68%, respectively. In this study, several kinds of poly-hydroxyalkanoates (PHAs) were observed in cells. These included 24% poly-3-hydroxybutyrate (PHB), 41% poly-3-hydroxyvalerate (PHV), 18% poly-3 hydroxyhexanoate (PHH), 10% poly-3-hydroxyoctanoate (PHO), 5% poly-3 hydroxydecanoate (PHD). and 2% poly-3-hydroxydodecanoate (PHDD), indicating that microorganisms could store various PHAs through the different metabolic pathways. However, breakdown of the enhanced biological phosphorus removal (EBPR) mechanism was observed when SRT increased from 30 to 50 days for the enhancement of nitrification. To study the effect of SRT on EBPR, a sequencing batch reactor (SBR) system that was supplied with glucose was operated at various SRTs of 5, 10, and 15 days. Nitrification and denitrification efficiencies increased as SRT increased. However, the content of intracellular materials such as PHAs, glycogen. and poly-P in cells decreased. From these results, it was concluded that SRT should be carefully controlled to increase nitrification activity and to maintain biological phosphorus removal activity in the BNR process. PMID- 15338700 TI - Of: Odor and volatile organic compound treatment by biotrickling filters: pilot scale studies at hyperion treatment plant, H. H. J. Cox, M. A. Deshusses, B. M. Converse, E. D. Schroeder, R. Iranpour 74, 557 (2002). PMID- 15338701 TI - Of: Factors affecting ballasted flocculation reactions, James C. Young and Findlay G. Edwards, 75, 263 (2003). PMID- 15338702 TI - Proposals to ban hunting: welfare implications for hunting dogs. PMID- 15338703 TI - Brucellosis outbreak: no evidence for further spread. PMID- 15338704 TI - Who will regulate the profession under a new Veterinary Surgeons Act? AB - A resolution by the Council of the Royal College of Veterinary Surgeons in March has set the College on the road towards achieving regulation for all groups of people providing veterinary services, not just veterinary surgeons. Here, Mr Roger Eddy, chairman of the RCVS working party on a new Veterinary Surgeons Act, discusses some of the options being considered by the College, and seeks views on an appropriate way forward PMID- 15338705 TI - Antibodies to selected pathogens in free-ranging terrestrial carnivores and marine mammals in Canada. AB - Antibody titres to selected pathogens (canine adenovirus [CAV-2], feline herpesvirus [FHV], phocine herpesvirus [PHV-1], canine distemper virus, dolphin morbillivirus [DMV], phocine distemper virus [PDV], parainfluenza virus type 3 [PI3], rabies virus, dolphin rhabdovirus [DRV], canine coronavirus, feline coronavirus, feline leukaemia virus, Borrelia burgdorferi and Toxoplasma gondii) were determined in whole blood or serum samples from selected free-ranging terrestrial carnivores and marine mammals, including cougars (Fellis concolor), lynxes (Fellis lynx), American badgers (Taxidea taxus), fishers (Martes pennanti), wolverines (Gulo gulo), wolves (Canis lupus), black bears (Ursus americanus), grizzly bears (Ursus arctos), polar bears (Ursus maritimus), walruses (Odobenus rosmarus) and belugas (Delphinapterus leucas), which had been collected at several locations in Canada between 1984 and 2001. Antibodies to a number of viruses were detected in species in which these infections have not been reported before, for example, antibodies to CAV-2 in walruses, to PDV in black bears, grizzly bears, polar bears, lynxes and wolves, to DMV in grizzly bears, polar bears, walruses and wolves, to PI3 in black bears and fishers, and to DRV in belugas and walruses. PMID- 15338706 TI - Associations between the prion protein genotype and performance traits of meat breeds of sheep. AB - The prion protein (PrP) genotypes of four German meat breeds of sheep were examined in relation to their scores for muscle mass, conformation, wool quality, daily liveweight gain and ultrasonic measurements of the depth of back muscle and back fat. The dataset included 912 genotyped German texel sheep among 10,383 recorded sheep, 474 genotyped Suffolk sheep among 4079 recorded sheep, 271 genotyped German white-headed mutton sheep among 3393 recorded sheep, and 99 genotyped German black-headed mutton sheep among 1642 recorded sheep. The model included the fixed effects of PrP genotype, herd, year and season, test day, sex, rearing method, the regression coefficient of average age, and the random additive genetic effect of the animal. In all four breeds there were no statistically significant associations between these performance traits and the occurrence of ARR alleles and the ARR/ARR genotype except for the depth of back muscle and the daily liveweight gain of German black-headed mutton sheep; however, in this breed, the interpretation could be affected by the small number of animals involved. All the genotyped sheep had significantly better performance traits than the non-genotyped sheep. PMID- 15338707 TI - Inheritance and determinants of pulmonary oedema in swedish hunting dogs. AB - By using information derived from questionnaires sent to registered owners of drever dams and sires in Sweden with offspring born in 1992 and/or 1994, two groups of offspring were identified: one with one parent said to have had breathing difficulties after hunting, and another with both parents unaffected. Questionnaires were sent to the owners of these offspring, and multiple logistic regression was used to analyse the offspring data, with the status of their sire and dam with respect to breathing difficulties after hunting included as covariates. For the outcome 'breathing difficulties after hunting' in the 266 offspring, the odds ratio (OR) was 4.4 (95 per cent confidence interval [CI] 1.8 to 10.8) if the dam was affected and 3-9 (95 per cent CI 1.2 to 11.1) if the sire was affected. The OR for male offspring was 2.4 (95 per cent CI 1.1 to 5.7). The heritability of the condition was estimated to be 0.34 from the dogs born in 1992, and 0.28 from the dogs born in 1994. PMID- 15338708 TI - Genetic similarity between porcine circovirus type 2 isolated from the first reported case of PMWS in South Africa and North American isolates. PMID- 15338709 TI - Metabolic homeostasis in postpartum dairy cows hampered by fatty livers. PMID- 15338710 TI - Fixation of femur, humerus and tibia fractures in cats using intramedullary threaded Steinmann pins. PMID- 15338711 TI - Slaughter without prestunning. PMID- 15338712 TI - Docking dogs' tails. PMID- 15338713 TI - Isolation of an avian paramyxovirus type 9 from migratory waterfowl in Italy. PMID- 15338714 TI - Possible risperidone poisoning in a dog. PMID- 15338715 TI - [Possibilities of complex etiological diagnosis of chronic viral hepatitides B and C]. AB - Etiological markers for HBV and HBC were studied in 26 cases of chronic hepatitis. In 21 cases coincidence of serological and immunohistochemical results was found, in 9 cases markers of other viruses were registered. Immunohistochemical markers were identified in 3 of 5 liver biopsies in patients with negative serological tests. Histological signs of HBV and HBC corresponded to those of serological and immunohistochemical ones in 17 cases. PMID- 15338716 TI - [Morphological features of chronic viral hepatitis in drug addicts]. AB - Liver pathology in parenteral drug abusers (PDA) was studied morphometrically. Chronic hepatitis in PDA is characterized by high incidence of lymphoid follicles formation in the portal tracts and intralobularly, relatively low activity of portal hepatitis in significant expression of the lobular component of the inflammatory process, marked fibrosis and early formation of micronodular liver cirrhosis, noticeable proliferation of narrow bile ducts, high content of neutrophils, eosinophils, siderophages and common macrophages in the infiltrate and their granuloma-like accumulations in the portal tracts. These criteria are recommended for diagnosis of drug abuse by morphological data. The use of a morphometrical method is proposed for studying morphological manifestations of chronic hepatitis, for perfection of their classification, in particular. PMID- 15338717 TI - [Ultrastructural changes of gastric superficial epithelium cells in chronic Helicobacter gastritis]. AB - Electron-microscopic investigations were carried out in 22 specimens of biopsy material from pyloric and fundal parts of the stomach in patients with chronic Helicobacter gastritis. In 6 cases of normal mucosa Helicobacter pylori (HP) was absent both in the antrum and fundus. In chronic antral gastritis HP was found in both regions. Variants of HP adhesion to the epithelium and its interactions were studied. It is demonstrated that HP may penetrate into intercellular spaces and cell cytoplasm. Cell reciprocal reaction to adhesion and invasion was accompanied by hyperplasia of the microvilli, clasmatosis and "selfsanation" phenomenon. Differences between ultrastructural changes in the antrum and fundus were considered. PMID- 15338718 TI - [Changes in the ultrastructure of the stomach mucous membrane parietal cells caused by inhibitors of hydrochloric acid secretion]. AB - The study of the action of phamotidine and omeprazol on the stomach parietal cells in patients with duodenal ulcer has shown that phamotidin results in changes of secretory membrane of the parietal cells increasing its secretory potential while omeprazol reduces energetic metabolism of the lining cell by the impact on its mitochondrial apparatus. Both in children and adults with duodenal ulcer more developed mitochondrial cell activity was found after omeprazol treatment. PMID- 15338719 TI - [Immunomorphological characteristics of the infiltrative growth in well differentiated thyroid carcinoma]. AB - An immunomorphological study of 52 samples of normal thyroid tissue, 10 adenomas, 42 well differentiated carcinomas and 2 metastases has been performed using markers of the epithelium, extracellular matrix and angiogenesis. Basal keratin N 17 of the compound epithelium occurred only in malignant tumors. The highest number of the keratin N 17-positive cells was registered in sclerosing A-cell cancer, medullary and mixed C-cell cancer which is very aggressive. These cells locate in the foci of proliferation of tumor structures and in the regions of infiltrative growth. Multiple defects of basal membranes and prevalence of capillaries marked by antibodies to CD-31 over those identified by the antibodies to factor VIII were found in the areas of strong expression of keratin 17. Thus, the above markers detect foci of aggressive growth even in well differentiated thyroid carcinomas. This fact may be of prognostic value in oncomorphological practice. PMID- 15338720 TI - [Expression of argyrophilic proteins from the nucleolar organizer regions as an index of maturity degree of benign and malignant adrenal tumors]. AB - Expression of APNOR was studied in 26 benign and malignant tumors of adrenal cortex and medulla. The histochemical method with silver nitrate was used. Expression of APNOR argyrophilic proteins in adrenocortical carcinomas was 4.59 times higher than in adrenocortical adenomas and 2.63 times higher in pheochromoblastomas than in pheochromocytomas. This index may be recommended as an additional method for differential diagnosis of benign and malignant adrenal tumors, particularly in difficult border line cases as well as determination of prognosis and metastatic potential of these tumors. Cytospecificity of APNOR expression in different types of adrenal cells was established. PMID- 15338721 TI - [Ploidometry in improving the quality of pathohistologic diagnosis]. AB - A computer image analyzer (imager-CG with a version of ploidometry program "Avtan San") was utilized for raising quality of differential diagnosis process in pathohistological examinations of biopsies. The results of ploidometry performed on histological slides stained by Felgen allowed to diagnose the degree of intraepithelial neoplasia and different stages of infiltrating carcinomas differentiation. The use of stereometric methods and immunocytohistometry for pathohistological diagnosis is described. PMID- 15338722 TI - [Dynamics of hemocirculatory bed changes in the wall of the dog small bowel after modeling acute strangulation ileum in experiment]. AB - It is shown that in acute experimental ileum microcirculatory disturbances arise earlier and disappear later than clinical manifestations. General chain of the strangulation ileum are microcirculatory changes in the intestinal wall. They are of one type, depend on the time of ileum and determine severity and reversibility of intestinal wall damage. PMID- 15338723 TI - Telomere biology and the molecular basis of aging. AB - Process of aging is regulated on the level of organism, clones and cellular level. Genes regulating aging on the individual (organism) level are detected in unicellular fungi. Mutations of such genes may lead to life lengthening by 60%. A similar gene is found in mice. Aging at the clonal level takes place by means of telomere shortening--that of DNA regions close to chromosome endings. Telomere shortening results in the chromosomes instability, their breaks and mutations, this apparently being a mechanism responsible for the increase of cancer incidence at advanced age. Telomerase is an enzyme responsible for the stability of the telomere length, supporting proliferation and by this that of life. This explains the increase of telomerase activity practically in all cases of malignant tumors. Aging is regulated by molecular components of cell cycle on cellular level. PMID- 15338724 TI - Synovial sarcoma (SS): new perspectives supported by modern technology. AB - Based upon the experience of 256 cases of synovial sarcoma (SS), the present review analyzes structural, biological and molecular pathology of this poorly known sarcoma. The histology displays a multiphenotype with two major components: biphasic and monophasic SS. In addition, a number of variants have been described: undifferentiated Ewing's like, myoxid and predominantly epithelial (monophasic epithelial sarcoma). Microcalcifications and squamous metaplasia are often seen in the tumor. Immunohistochemistry with EMA and cytokeratin in the epithelial or epithelioid component is diagnostic for SS together with vimentin positivity in the spindle cells. Several other epitopes are also expressed (CD99, CD56, C-MET, HGF/SF, CD44). The ultrastructure confirms the variegated pattern of the neoplasm demonstrating the epithelial component and the epithelioid or spindle cell type closely associated with each other. Transition of epithelial cells to epithelioid and spindle-like mesenchymal component is seen. Nude-mice xenografts and cell lines after in vitro culture confirm heterogeneity of this sarcoma. Molecular histology of the SS has provided high utility not only for their differential diagnosis due to a specific chromosomal translocation: t(X;18)(p11.2;q11.2) but also after cloning these breakpoints resulting in the fusion of two genes: SYT at 18q11 and SSX at Xp11. Further observations have lead to distinguish the existence of two related genes: SSX1 and SSX2, that provide a highly specific and sensitive diagnostic marker for SS. Moreover, clinical correlations have demonstrated that SYT-SSX1 leads to a poor clinical outcome while the fusion SYT-SSX2 provides survival advantages to the patients. PMID- 15338725 TI - The biological role and regulation of matrix metalloproteinases (MMP) in cancer. AB - Matrix metalloproteinases (MMP) are zinc-dependent proteolytic enzymes capable of breaking down basement membranes and most extracellular matrix (ECM) components. MMP expression and activation are carefully regulated in physiological conditions in order to prevent uncontrolled destruction of body tissues but this regulation is modified or disrupted in pathological processes, including cancer. This review presents regulatory mechanisms designed to control MMP action. These consist of direct activation and inhibition by tissue inhibitors of MMP (TIMP), signal transduction pathways mediated by adhesion molecules such as integrins and EMMPRIN, involved in activation of MMP synthesis and transcriptional control by the ETS family of transcription factors. PMID- 15338726 TI - [Serous papillary adenocarcinoma of the endometrium]. AB - Serous-papillary endometrial adenocarcinoma (SPEA) accounts for 10% of endometrial carcinoma. Histological similarity with ovarian carcinoma originating from the cylioepithelial cyst is explained by the same histogenesis from Muller epithelium analogs. The description is given of SPEA with essential content of psammotic bodies, embolism and development of intramural, cervical, parametral and lymphogenic metastases. A relatively benign clinical picture of SPEA psammotic variant does not correlate with aggressiveness of the tumor growth. PMID- 15338727 TI - A constrained variational principle for direct estimation and smoothing of the diffusion tensor field from complex DWI. AB - In this paper, we present a novel constrained variational principle for simultaneous smoothing and estimation of the diffusion tensor field from complex valued diffusion-weighted images (DWI). The constrained variational principle involves the minimization of a regularization term of L(P) norms, subject to a nonlinear inequality constraint on the data. The data term we employ is the original Stejskal-Tanner equation instead of the linearized version usually employed in literature. The complex valued nonlinear form leads to a more accurate (when compared to the linearized version) estimate of the tensor field. The inequality constraint requires that the nonlinear least squares data term be bounded from above by a known tolerance factor. Finally, in order to accommodate the positive definite constraint on the diffusion tensor, it is expressed in terms of Cholesky factors and estimated. The constrained variational principle is solved using the augmented Lagrangian technique in conjunction with the limited memory quasi-Newton method. Experiments with complex-valued synthetic and real data are shown to depict the performance of our tensor field estimation and smoothing algorithm. PMID- 15338728 TI - Neighbor-constrained segmentation with level set based 3-D deformable models. AB - A novel method for the segmentation of multiple objects from three-dimensional (3 D) medical images using interobject constraints is presented. Our method is motivated by the observation that neighboring structures have consistent locations and shapes that provide configurations and context that aid in segmentation. We define a maximum a posteriori (MAP) estimation framework using the constraining information provided by neighboring objects to segment several objects simultaneously. We introduce a representation for the joint density function of the neighbor objects, and define joint probability distributions over the variations of the neighboring shape and position relationships of a set of training images. In order to estimate the MAP shapes of the objects, we formulate the model in terms of level set functions, and compute the associated Euler Lagrange equations. The contours evolve both according to the neighbor prior information and the image gray level information. This method is useful in situations where there is limited interobject information as opposed to robust global atlases. In addition, we compare our level set representation of the object shape to the point distribution model. Results and validation from experiments on synthetic data and medical imagery in two-dimensional and 3-D are demonstrated. PMID- 15338729 TI - Genus zero surface conformal mapping and its application to brain surface mapping. AB - We developed a general method for global conformal parameterizations based on the structure of the cohomology group of holomorphic one-forms for surfaces with or without boundaries (Gu and Yau, 2002), (Gu and Yau, 2003). For genus zero surfaces, our algorithm can find a unique mapping between any two genus zero manifolds by minimizing the harmonic energy of the map. In this paper, we apply the algorithm to the cortical surface matching problem. We use a mesh structure to represent the brain surface. Further constraints are added to ensure that the conformal map is unique. Empirical tests on magnetic resonance imaging (MRI) data show that the mappings preserve angular relationships, are stable in MRIs acquired at different times, and are robust to differences in data triangulation, and resolution. Compared with other brain surface conformal mapping algorithms, our algorithm is more stable and has good extensibility. PMID- 15338730 TI - Estimation of the hemodynamic response in event-related functional MRI: Bayesian networks as a framework for efficient Bayesian modeling and inference. AB - A convenient way to analyze blood-oxygen-level-dependent functional magnetic resonance imaging data consists of modeling the whole brain as a stationary, linear system characterized by its transfer function: the hemodynamic response function (HRF). HRF estimation, though of the greatest interest, is still under investigation, for the problem is ill-conditioned. In this paper, we recall the most general Bayesian model for HRF estimation and show how it can beneficially be translated in terms of Bayesian graphical models, leading to 1) a clear and efficient representation of all structural and functional relationships entailed by the model, and 2) a straightforward numerical scheme to approximate the joint posterior distribution, allowing for estimation of the HRF, as well as all other model parameters. We finally apply this novel technique on both simulations and real data. PMID- 15338731 TI - Object-based morphometry of the cerebral cortex. AB - Most of the approaches dedicated to automatic morphometry rely on a point-by point strategy based on warping each brain toward a reference coordinate system. In this paper, we describe an alternative object-based strategy dedicated to the cortex. This strategy relies on an artificial neuroanatomist performing automatic recognition of the main cortical sulci and parcellation of the cortical surface into gyral patches. A set of shape descriptors, which can be compared across subjects, is then attached to the sulcus and gyrus related objects segmented by this process. The framework is used to perform a study of 142 brains of the International Consortium for Brain Mapping (ICBM) database. This study reveals some correlates of handedness on the size of the sulci located in motor areas, which was not detected previously using standard voxel based morphometry. PMID- 15338732 TI - Performance-based classifier combination in atlas-based image segmentation using expectation-maximization parameter estimation. AB - It is well known in the pattern recognition community that the accuracy of classifications obtained by combining decisions made by independent classifiers can be substantially higher than the accuracy of the individual classifiers. We have previously shown this to be true for atlas-based segmentation of biomedical images. The conventional method for combining individual classifiers weights each classifier equally (vote or sum rule fusion). In this paper, we propose two methods that estimate the performances of the individual classifiers and combine the individual classifiers by weighting them according to their estimated performance. The two methods are multiclass extensions of an expectation maximization (EM) algorithm for ground truth estimation of binary classification based on decisions of multiple experts (Warfield et al., 2004). The first method performs parameter estimation independently for each class with a subsequent integration step. The second method considers all classes simultaneously. We demonstrate the efficacy of these performance-based fusion methods by applying them to atlas-based segmentations of three-dimensional confocal microscopy images of bee brains. In atlas-based image segmentation, multiple classifiers arise naturally by applying different registration methods to the same atlas, or the same registration method to different atlases, or both. We perform a validation study designed to quantify the success of classifier combination methods in atlas based segmentation. By applying random deformations, a given ground truth atlas is transformed into multiple segmentations that could result from imperfect registrations of an image to multiple atlas images. In a second evaluation study, multiple actual atlas-based segmentations are combined and their accuracies computed by comparing them to a manual segmentation. We demonstrate in both evaluation studies that segmentations produced by combining multiple individual registration-based segmentations are more accurate for the two classifier fusion methods we propose, which weight the individual classifiers according to their EM based performance estimates, than for simple sum rule fusion, which weights each classifier equally. PMID- 15338733 TI - Principal geodesic analysis for the study of nonlinear statistics of shape. AB - A primary goal of statistical shape analysis is to describe the variability of a population of geometric objects. A standard technique for computing such descriptions is principal component analysis. However, principal component analysis is limited in that it only works for data lying in a Euclidean vector space. While this is certainly sufficient for geometric models that are parameterized by a set of landmarks or a dense collection of boundary points, it does not handle more complex representations of shape. We have been developing representations of geometry based on the medial axis description or m-rep. While the medial representation provides a rich language for variability in terms of bending, twisting, and widening, the medial parameters are not elements of a Euclidean vector space. They are in fact elements of a nonlinear Riemannian symmetric space. In this paper, we develop the method of principal geodesic analysis, a generalization of principal component analysis to the manifold setting. We demonstrate its use in describing the variability of medially-defined anatomical objects. Results of applying this framework on a population of hippocampi in a schizophrenia study are presented. PMID- 15338734 TI - Constructing diffeomorphic representations for the groupwise analysis of nonrigid registrations of medical images. AB - Groupwise nonrigid registrations of medical images define dense correspondences across a set of images, defined by a continuous deformation field that relates each target image in the group to some reference image. These registrations can be automatic, or based on the interpolation of a set of user-defined landmarks, but in both cases, quantifying the normal and abnormal structural variation across the group of imaged structures implies analysis of the set of deformation fields. We contend that the choice of representation of the deformation fields is an integral part of this analysis. This paper presents methods for constructing a general class of multi-dimensional diffeomorphic representations of deformations. We demonstrate, for the particular case of the polyharmonic clamped-plate splines, that these representations are suitable for the description of deformations of medical images in both two and three dimensions, using a set of two-dimensional annotated MRI brain slices and a set of three-dimensional segmented hippocampi with optimized correspondences. The class of diffeomorphic representations also defines a non-Euclidean metric on the space of patterns, and, for the case of compactly supported deformations, on the corresponding diffeomorphism group. In an experimental study, we show that this non-Euclidean metric is superior to the usual ad hoc Euclidean metrics in that it enables more accurate classification of legal and illegal variations. PMID- 15338735 TI - Tracheal and central bronchial aerodynamics using virtual bronchoscopy and computational fluid dynamics. AB - Virtual bronchoscopy reconstructions of the airway noninvasively provide useful morphologic information of structural abnormalities such as stenoses and masses. In this paper, we show how virtual bronchoscopy can be used to perform aerodynamic calculations in anatomically realistic models. Pressure and flow patterns in a human airway were computed noninvasively. These showed decreased pressure and increased shear stress in the region of a stenosis. PMID- 15338736 TI - Adaptive averaging for improved SNR in real-time coronary artery MRI. AB - A technique has been developed for combining a series of low signal-to-noise ratio (SNR) real-time magnetic resonance (MR) images to produce composite images with high SNR and minimal artifact in the presence of motion. The main challenge is identifying a set of real-time images with sufficiently small systematic differences to avoid introducing significant artifact into the composite image. To accomplish this task, one must: 1) identify images identical within the limits of noise; 2) detect systematic errors within such images with sufficient sensitivity. These steps are achieved by evaluating the correlation coefficient (CC) between regions in prospective images and a template containing the anatomy of interest. Images identical within noise are selected by comparing the measured CC values to the theoretical distribution expected due to noise. Sensitivity for systematic error depends on the SNR of the CC (=SNR(CCmax)), which in turn depends on the noise, and the template size and structure. By varying the template size, SNR(CCmax) may be altered. Experiments on phantoms and coronary artery images demonstrate that the SNR(CCmax) necessary to avoid introducing significant artifact varies with the target composite SNR. The future potential of this technique is demonstrated on high-resolution (approximately 0.9 mm), reduced field-of-view real-time coronary images. PMID- 15338737 TI - Respiratory motion of the heart from free breathing coronary angiograms. AB - Respiratory motion compensation for cardiac imaging requires knowledge of the heart's motion and deformation during breathing. This paper presents a method for measuring the natural tidal respiratory motion of the heart from free breathing coronary angiograms. A three-dimensional (3-D) deformation field describing the cardiac and respiratory motion of the coronary arteries is recovered from a biplane acquisition. A cardiac respiratory parametric model is formulated and used to decompose the deformation field into cardiac and respiratory components. Angiograms from ten patients were analyzed. A 3-D translation motion model was sufficient for describing the motion of the heart in only two patients. For all patients, the heart translated caudally (mean, 4.9+/-1.9 mm; range, 2.4 to 8.0 mm) and underwent a cranio-dorsal rotation (mean, 1.5 degrees+/-0.9 degrees; range, 0.2 degrees to 3.5 degrees) during inspiration. In eight patients, the heart also translated anteriorly (mean, 1.3+/-1.8 mm; range, -0.4 to 5.1 mm) and rotated in a caudo-dextral direction (mean, 1.2 degrees+/-1.3 degrees; range, 1.9 degrees to 3.2 degrees). PMID- 15338738 TI - Thymidylate synthase inhibitors. PMID- 15338739 TI - Anthracyclines. PMID- 15338740 TI - Tubulin-targeting agents. PMID- 15338741 TI - Topoisomerase I interactive agents. AB - Increased insight into the mechanism of interaction of topoisomerase I interactive agents will maximize the therapeutic index and enhance the development of additional agents. Preclinical studies designed to elucidate mechanisms by which the topoisomerase I interactive agents induce cell death will be essential. The role of ABC transporters in resistance to topoisomerase I interactive agents has been recently appreciated and future studies should be directed at circumventing this resistance. The results of preclinical studies must be translated into the design of clinical trials so that these agents can be used rationally. In this regard results of preclinical studies have clearly pointed to the enhanced antitumor activity from protracted dosing of topoisomerase I interactive agents and results of clinical trials are now supporting these preclinical findings. Finally, investigators are trying to understand better the mechanism(s) of the dose-limiting toxicities observed with the currently available topoisomerase I interactive agents in an effort to enable the optimal dosing of these agents. Even though the first priority must be to determine the therapeutic potential of the currently available agents, it is reassuring to know that other topoisomerase I interactive agents are currently under development. PMID- 15338742 TI - Topoisomerase II inhibitors. PMID- 15338743 TI - Farnesyltransferase inhibitors. PMID- 15338744 TI - Cyclin-dependent kinases as targets for cancer therapy. AB - Cell cycle perturbations are commonly observed in human malignancies. Exploiting this finding is the rationale for the development of CDK inhibitors as anti-tumor agents. Single-agent evaluation of several CDKIs has demonstrated limited clinical activity. The combination of CDKIs with standard cytotoxic agents is an emerging, alternative approach to anticancer therapy that also exploits the cell cycle perturbations of malignancy. Pre-clinical studies demonstrate the concept of cell cycle mediated drug resistance, and suggest that the combination of standard cytotoxic agents with CDKIs will require thoughtful sequencing and scheduling. With this in mind, there are presently several clinical investigations underway examining the combination of a standard cytotoxic with a novel CDKI, with particular attention to sequence and scheduling. Although phase II evaluation of these combination studies will provide initial evidence of anti tumor activity, definitive phase III studies will be needed to establish this class of agents in the care of patients with cancer. PMID- 15338745 TI - DNA-binding agents. PMID- 15338746 TI - Advances in endocrine therapy for the treatment and prevention of breast cancer. PMID- 15338747 TI - Monoclonal antibody therapy for breast cancer: herceptin. PMID- 15338748 TI - Rituximab. PMID- 15338749 TI - Mucins in gastrointestinal cancers. AB - The mucin family has been under study by molecular biologists, biochemists, pathologists and immunologists interested in cancer because of the role these molecules can play in the diagnosis and treatment of cancer. Immense knowledge has been accumulated, but the high speed of progress in the laboratory has not been matched by the progress towards applying this knowledge in the clinic. For example, specific knowledge of cancer-associated changes in the expression and glycosylation of various mucins, which can aid in the diagnosis as well as prognosis of GI cancers, has not yet led to the use of a panel of anti-mucin antibodies as a standard diagnostic tool. Similarly, many more opportunities exist for using mucin-based therapies than are currently being considered in the clinic. This chapter aimed to highlight some of these opportunities and to interest clinician scientists in exploring them in the near future. PMID- 15338750 TI - HER-2/neu vaccines. PMID- 15338751 TI - Colon cancer antigen and anti-idiotype vaccines. PMID- 15338752 TI - Anti-carcinoembryonic antigen immunity. PMID- 15338753 TI - Cancer gene therapy and immunotherapy. PMID- 15338754 TI - Recent areas of development for dendritic cell vaccines. PMID- 15338755 TI - Mechanisms of tumor evasion from the immune response. AB - The results from in vitro immunological experiments, murine tumor models and patients with cancer clearly demonstrate that tumors have multiple mechanisms to evade the immune response. During the early stages of tumor development malignant cells can be poor stimulators, present poor targets or become resistant to the innate immune response, while at later stages, progressively growing tumors impair the adaptive immune response by blocking the maturation and function of APCs and causing alterations in T-cell signal transduction and function. Preliminary results also suggest a correlation between some of these changes and an increased metastatic potential of the tumor cells, a diminished response to immunotherapy, and poor prognosis. Carefully coordinated basic research studies and clinical immunotherapy trials will be required to fully determine the impact of these mechanisms of tumor evasion on the outcome of the disease and the response to treatment. However, understanding the mechanisms used by tumor cells to evade the immune system could result in new therapeutic approaches for preventing and/or reversing these immune alterations and could have the potential of improving the current results of immunotherapy trials. PMID- 15338756 TI - Plasma cell myeloma and leukemia. PMID- 15338757 TI - Lymphomas. PMID- 15338758 TI - Head and neck cancers. PMID- 15338759 TI - Lung cancer. PMID- 15338760 TI - Upper gastrointestinal tumors. PMID- 15338761 TI - Cancers of the large bowel and hepatobiliary tract. PMID- 15338762 TI - Neuroendocrine tumors--somatostatin receptor expression and somatostatin analog treatment. PMID- 15338763 TI - Genitourinary malignancies. PMID- 15338764 TI - Systemic therapy for gynecological neoplasms: ovary, cervix and endometrium. AB - Early-stage endometrial carcinomas should be treated by total abdominal hysterectomy and bilateral salpingo-oophorectomy. In a small subset of patients, who wish to have children, conservative treatment (hormonal therapy) could be considered. The most effective agents for palliation of advanced disease are doxorubicin plus cisplatin. PMID- 15338765 TI - Breast cancer. PMID- 15338766 TI - Melanoma--new developments in diagnosis and therapy. PMID- 15338767 TI - Sarcoma. PMID- 15338768 TI - Brain tumors. PMID- 15338769 TI - Biology and therapy of malignant solid tumors in childhood. PMID- 15338770 TI - Supportive care. PMID- 15338771 TI - AIDS-associated malignancies. PMID- 15338772 TI - Risk assessment and screening for colorectal cancer. PMID- 15338773 TI - Non-steroidal anti-inflammatory drugs (NSAIDs) for colorectal cancer prevention. PMID- 15338774 TI - Molecular predictors of prognosis and response to therapy in colorectal cancer. PMID- 15338775 TI - Combined modality therapy for rectal cancer. PMID- 15338776 TI - New drugs for colorectal cancer. PMID- 15338777 TI - Liver directed therapies for colorectal cancer. PMID- 15338778 TI - Suicide in schizophrenic patients: a neglected issue. PMID- 15338779 TI - Bradley method offers option for natural childbirth. PMID- 15338780 TI - Emergency contraception: an ongoing debate. PMID- 15338781 TI - Allergen immunotherapy. AB - Allergen immunotherapy (also called allergy vaccine therapy) involves the administration of gradually increasing quantities of specific allergens to patients with IgE-mediated conditions until a dose is reached that is effective in reducing disease severity from natural exposure. The major objectives of allergen immunotherapy are to reduce responses to allergic triggers that precipitate symptoms in the short term and to decrease inflammatory response and prevent development of persistent disease in the long term. Allergen immunotherapy is safe and has been shown to be effective in the treatment of stinging-insect hypersensitivity, allergic rhinitis or conjunctivitis, and allergic asthma. Allergen immunotherapy is not effective in the treatment of atopic dermatitis, urticaria, or headaches and is potentially dangerous if used for food or antibiotic allergies. Safe administration of allergen immunotherapy requires the immediate availability of a health care professional capable of recognizing and treating anaphylaxis. An observation period of 20 to 30 minutes after injection is mandatory. Patients should not be taking beta-adrenergic blocking agents when receiving immunotherapy because these drugs may mask early signs and symptoms of anaphylaxis and make the treatment of anaphylaxis more difficult. Unlike antiallergic medication, allergen immunotherapy has the potential of altering the allergic disease course after three to five years of therapy. PMID- 15338782 TI - Information from your family doctor. Allergy shots-what you need to know. PMID- 15338783 TI - Emergency contraception. AB - Women can use emergency contraception to prevent pregnancy after known or suspected failure of birth control or after unprotected intercourse. Many patients do not ask for emergency contraception because they do not know of its availability. Emergency contraception has been an off-label use of oral contraceptive pills since the 1960s. Dedicated products, the Yuzpe regimen (Preven) and levonorgestrel (Plan B), were marketed in the United States after 1998 but had been available in Europe for years before that. A third approved method of emergency contraception is the insertion of an intrauterine device. Emergency contraception is about 75 to 85 percent effective. It is most effective when initiated within 72 hours after unprotected intercourse. The mechanism of action may vary, depending on the day of the menstrual cycle on which treatment is started. Despite the large number of women who have received emergency contraception, there have been no reports of major adverse outcomes. If a woman becomes pregnant after using emergency contraception, she may be reassured about the lack of negative effects emergency contraception has on fetal development. It may be beneficial for physicians to offer an advance prescription for emergency contraception at a patient's regular gynecologic visit to help reduce unwanted pregnancies. Advance provision of emergency contraception can increase its use significantly without adversely affecting the use of routine contraception. PMID- 15338784 TI - Information from your family doctor. Emergency birth control. PMID- 15338785 TI - Making decisions with families at the end of life. AB - Because advance directives are not yet the norm, end-of-life decisions for patients without medical decision-making capacity are made regularly within discussions between the patient's physician and family. Communication and decision making in these situations require a complex integration of relevant conceptual knowledge of ethical implications, the principle of surrogate decision making, and legal considerations; and communication skills that address the highly charged emotional issues under discussion. The most common pitfalls in establishing plans of care for patients who lack decision-making capacity include failure to reach a shared appreciation of the patient's condition and prognosis; failure to apply the principle of substituted judgment; offering the choice between care and no care, rather than offering the choice between prolonging life and quality of life; too literal an interpretation of an isolated, out-of context, patient statement made earlier in life; and failure to address the full range of end-of-life decisions from do-not-resuscitate orders to exclusive palliative care. PMID- 15338786 TI - Information from your family doctor. End-of-Life choices for families. PMID- 15338788 TI - Information from your family doctor. Colic: what you should know. PMID- 15338787 TI - Infantile colic. AB - Infantile colic can be distressing to parents whose infant is inconsolable during crying episodes. Colic is often defined by the "rule of three": crying for more than three hours per day, for more than three days per week, and for longer than three weeks in an infant who is well-fed and otherwise healthy. The physician's role is to ensure that there is no organic cause for the crying, offer balanced advice on treatments, and provide support to the family. Colic is a diagnosis of exclusion that is made after performing a careful history and physical examination to rule out less common organic causes. Treatment is limited. Feeding changes usually are not advised. Medications available in the United States have not been proved effective in the treatment of colic, and most behavior interventions have not been proved to be more effective than placebo. Families may turn to untested resources for help, and the physician should offer sound advice about these treatments. Above all, parents need reassurance that their baby is healthy and that colic is self-limited with no long-term adverse effects. Physicians should watch for signs of continuing distress in the child and family, particularly in families whose resources are strained already. PMID- 15338789 TI - Skin fragility of the hands. PMID- 15338790 TI - The efficacy of smoking cessation strategies in people with chronic obstructive pulmonary disease: results from a systematic review. AB - Smoking cessation is the most effective way to reduce the risk of developing chronic obstructive pulmonary disease (COPD). It prevents or delays the development of airflow limitation and also reduces its progression. The objective of this study was to systematically review the effects of interventions for smoking cessation in people with COPD. Comprehensive searches of electronic and internet databases were carried out from 1966 to March 2002, using the Cochrane Airways Group search strategy. The reference lists of all selected randomized trials and relevant reviews were inspected for additional published reports and citations of unpublished research. We evaluated the efficacy of behavioural interventions (e.g. counselling), pharmacotherapy (nicotine replacement therapy and non-nicotine therapy such as bupropion), and combinations of both. The main clinical outcome measure was prolonged abstinence after at least 6 months, confirmed by a biochemical test. Five trials comprising 6491 patients with COPD were included. Results of the Lung Health Study show that, by using an intensive behavioural (relapse prevention) programme combined with nicotine replacement therapy, prolonged abstinence rates are not only significantly higher compared with no intervention, but the difference in efficacy was sustained for over 5 years. A 12-week treatment course with bupropion sustained release combined with individual counselling, however, did not result in significantly higher prolonged abstinence rates after 12 months. Present evidence suggests that the most effective intervention for prolonged smoking cessation in patients with COPD is the combination of nicotine replacement therapy, coupled with an intensive, prolonged relapse prevention programme. PMID- 15338791 TI - Relief of dyspnoea by beta2-agonists after methacholine-induced bronchoconstriction. AB - Virtually all asthma patients use brorichodilators. Formoterol and salbutamol have a rapid onset of bronchodilating effect, whereas salmeterol acts slower. We studied the onset of improvement of dyspnoea sensation after inhalation with these bronchodilators and placebo to reverse a methacholine-induced bronchoconstriction as a model for an acute asthma attack. Seventeen patients with asthma completed this randomised, double-blind, crossover, double-dummy study. On 4 test days, forced expiratory volume in 1 s (FEV1) and Borg score were recorded and patients were challenged with methacholine until FEV1 fell with > or = 30% of baseline value. Thereafter, formoterol 12 microg via Turbuhaler, salbutamol 50 microg via Turbuhaler, salmeterol 50 microg via Diskhaler, or placebo was inhaled. FEV1 and Borg scores were assessed during the following 60 min. The first sensed improvement of Borg score was significantly (P<0.05) faster achieved with formoterol (geometric mean (Gmean) (range) 1.5 (1-40) min) and salbutamol 1.8 (1-10) min than with salmeterol 4.5 (1-30) min and placebo 3.4 (1 40) min. The Borg score returned significantly faster to the baseline value with formoterol, salbutamol, and salmeterol (Gmean time 13.8 (1-75), 13.4 (1-60), and 18.0 (1-75) min, respectively) than with placebo (33.6 (1-75 min). Formoterol and salbutamol act significantly faster than salmeterol in relieving dyspnoea induced by methacholine-induced bronchoconstriction, in patients with asthma. PMID- 15338792 TI - Diesel exhaust exposure enhances the expression of IL-13 in the bronchial epithelium of healthy subjects. AB - Epidemiological studies have demonstrated adverse health effects of environmental pollution. Diesel exhaust (DE) is an important contributor to ambient particulate matter pollution. DE exposure has been shown to induce a pronounced inflammatory response in the airways, with an enhanced epithelial expression of IL-8, and Gro alpha in healthy subjects. The present investigation was aimed to further characterise the epithelial response to DE in vivo, with particular reference to possible TH2 response, in non-atopic healthy subjects. To determine this response, 15 healthy, non-atopic non-smoking subjects with normal lung function were exposed to DE (PM10 300 microg/m3) and filtered air during 1 h on two separate randomised occasions. Bronchoscopy sampling of bronchial mucosal biopsies was performed 6 h after exposure. Immunohistochemical staining were performed using mAb for IL-10, IL-13 and IL-18 expression. DE exposure induced a significant increase in the expression of IL-13 in the bronchial epithelium cells, 2.1 (1.35-4.88) Md (Q1-Q3) vs. air 0.94 (0.53-1.23); P = 0.009. No significant changes were seen in IL-10 and IL-18 expression. This finding suggests an TH2-inflammatory response in the airways of non-atopic healthy individuals. PMID- 15338793 TI - Bronchial hyperreactivity and spirometric impairment in patients with seasonal allergic rhinitis. AB - We previously demonstrated in a group of patients with perennial allergic rhinitis alone, impairment of spirometric parameters and high percentage of bronchial hyperreactivity (BHR). Thus, the present study aimed at evaluating a group of subjects suffering from seasonal allergic rhinitis alone to investigate the presence of spirometric impairment and BHR both during and outside the pollen season. METHODS: One-hundred rhinitics sensitized to pollen allergens only were evaluated during and outside the pollen season. Spirometry and methacholine bronchial challenge were performed. RESULTS: Four rhinitics showed impaired values of FEV1 without referred symptoms of asthma during the pollen season. FEF 25-75 values were impaired in 17 rhinitics during the pollen season and in 11 rhinitics outside the pollen season (P<0.05). Fifty-four patients showed positive methacholine bronchial challenge both during and outside the pollen season. PD20/FEV1 methacholine was lower during the pollen season than outside (P<0.05). In BHR positive patients, reduced values of FVC (P<0.05), FEV1 (P<0.05), and FEF 25-75 (P<0.01) were significantly demonstrated in comparison with BHR negative rhinitics. There was a relationship between BHR degree and FEF 25-75 values only during the pollen season (P<0.001). CONCLUSIONS: This study evidences that an impairment of spirometric parameters may be observed also in patients with seasonal allergic rhinitis alone during the pollen season. A high percentage of these patients had BHR. A close relationship between upper and lower airways is confirmed also in the model of pollen allergy. Thus, a careful evaluation of lower airways should be performed also in those patients with seasonal allergic rhinitis alone. PMID- 15338794 TI - The dissemination of the British Guideline on the Management of Asthma 2003. AB - The BTS/SIGN British Guideline on the Management of Asthma was published in February 2003 (4). If health outcomes are to be influenced successfully, dissemination of the guideline and implementation of recommendations is essential. We report the dissemination activities undertaken during the 18 weeks following the guideline launch. To facilitate implementation a range of educational materials were produced reflecting the key messages from the guideline. In addition to postal mailing of the guideline to appropriate healthcare professionals, both educational materials and the guidelines were made freely available from the BTS and SIGN websites. In total, 135,710 copies of the guideline and 90,198 copies of the Quick Reference Guide were downloaded in the first 18 weeks, representing a considerable increase over the number of copies of the 1997 guidelines disseminated by mailing alone. Large quantities of educational materials were downloaded with many used for teaching purposes. An on line survey suggested that most respondents rated the materials as useful or very useful. Using websites to disseminate guidelines is a cost-effective method of informing health professionals of their content and is a more active process than the passive receipt of mailed copies. The availability of interactive educational materials for use in teaching appears to have been popular. PMID- 15338795 TI - The bronchodilatory effect of deep inspiration diminishes with aging. AB - Deep inspirations have the ability to dilate constricted airways. The impairment of this function has been associated with the occurrence of asthmatic symptoms. We evaluated whether the bronchodilatory effect of deep inspiration (DI) is affected by aging. We tested 25 healthy subjects (median age: 54 yrs, range: 25 83 yrs). Single dose methacholine (Mch) provocations were performed in the absence of DI, which induced at least 15% reduction in inspiratory vital capacity (IVC) from baseline. The post-Mch IVC measurement was followed by 4 DIs and by another IVC (post-DI IVC). The fractional difference between post-DI IVC and post Mch IVC represented the % bronchodilation by DI. The % bronchodilation significantly diminished with aging (r=0.65, P=0.0005). The bronchodilatory ability of DI was also positively associated with the degree of Mch-induced reduction in IVC (r=0.84, P<0.0001). In multiple regression analysis, where % bronchodilation was the dependent variable, both % reduction in IVC (P<0.0001) and age (P=0.02) entered the model. Our data raise the hypothesis that aging is associated with reduction in DI-induced bronchodilation. PMID- 15338796 TI - Tremor side effects of salbutamol, quantified by a laser pointer technique. AB - OBJECTIVE: To study tremor side effects of salbutamol an easily applicable, quick and low-priced method is needed. A new method using a commercially available, pen shaped laser pointer was developed. Aim of the study was to determine sensitivity, reproducibility, reference values and the agreement with a questionnaire. METHODS: Tremor was measured using a laser pointer technique. To determine sensitivity we assessed tremor in 44 patients with obstructive lung disease after administration of cumulative doses of salbutamol. Subjects were asked to aim at the centre of a target, subdivided in concentric circles, from 5 m distance. The circle in which the participant succeeded to aim was recorded in millimetres radius. In another series of measurements, reproducibility and reference values of the tremor was assessed in 65 healthy subjects in three sessions, at 9 a.m., 4 p.m. and 9 a.m., respectively, 1 week later. Postural tremor was measured with the arm horizontally outstretched rest tremor with the arm supported by an armrest and finally tremor was measured after holding a 2-kg weight until exhaustion. Inter-observer variability was measured in a series of 10 healthy subjects. Tremor was measured simultaneously by two independent observers. RESULTS: Salbutamol significantly increased tremor severity in patients in a dose-dependent way. Within healthy adults no age-dependency could be found (b = 0.262 mm/year; P = 0.72). There was no agreement between the questionnaire and tremor severity (r = 0.093; P = 0.53). Postural tremor showed no significant difference between the first and third session (P = 0.07). Support of the arm decreased tremor severity, exhaustion increased tremor severity significantly. A good agreement was found between two independent observers (interclass correlation coefficient 0.72). DISCUSSION: Quantifying tremor by using an inexpensive laser pointer is, with the exception of children (<12 years) a sensitive and reproducible method. PMID- 15338797 TI - Paradoxical results in the study of risk factors of chronic obstructive pulmonary disease (COPD) re-admission. AB - We have previously reported an apparently paradoxical association between medical care related factors and an increased risk of chronic obstructive pulmonary disease (COPD) re-hospitalisation, in a cohort of 346 COPD subjects from Barcelona, Spain. Confounding by severity or by indication is a plausible explanation. We tested the confounding effect of severity-related variables on these paradoxical associations. Forced expiratory volume in one second (FEV1), arterial oxygen pressure (PO2) and previous COPD admissions were associated with: (1) the presence of medical care related factors, and (2) re-admission during follow-up. Risks of readmission associated with most of the medical care related factors were reduced after adjustment for the severity variables. The risk associated with long-term oxygen therapy use changed from a crude OR of 2.36 (95% CI: 1.79-3.11) to an adjusted OR of 1.38 (0.95-2.00), while that associated with anticholinergics use varied from 3.52 (2.37-5.21) to 2.10 (1.32-3.36)). We concluded that the excess risk of COPD re-admission associated with medical care related factors might be partially due to confounding by indication. Residual confounding may still account for part of the remaining excess risk. True adverse effects of some pharmacological treatments cannot be excluded. PMID- 15338798 TI - Increased IgE-antibodies to Staphylococcus aureus enterotoxins in patients with COPD. AB - Recent evidence suggests that Staphylococcus aureus enterotoxins (SAEs) could modify airway disease by acting as superantigens, an immune response that can be monitored by detection of IgE antibodies to SAEs. We studied the expression of total IgE and specific IgE to SAEs using the Uni-CAP system in healthy controls, smokers without COPD and COPD patients. Only 1/10 controls (10%) and 1/16 smokers (6.3%) had IgE to SAEs compared to 7/18 patients with stable COPD (38.9%) and 21/54 patients with exacerbated COPD (38.9%). The IgE levels to SAEs of the patients with stable COPD (0.18 [0.05-26.2]kUA/l) and the patients with exacerbated COPD (0.09 [0.05-18.6]kUA/l) were significantly higher than those of smokers (n = 16; 0.05 [0.05-0.82]kUA/l) and controls (n = 11; 0.05 [0.05 0.9]kUA/l, P<0.05). IgE to SAEs decreased significantly in the exacerbated patients during hospitalization (0.13 [0.05-18.3] vs. 0.05 [0.05-11]kUA/l, P<0.001) going along with a significant increase in FEV1 (38.1 [16.9-79.5] vs. 51.6 [15-80]%predicted, P<0.001). Similarly to severe asthma, we found significantly elevated IgE to SAE in COPD patients. Our data for the first time suggest differences between healthy subjects, smokers and patients with established COPD regarding the role of bacterial products and point to a possible disease modifying role of SAEs. PMID- 15338799 TI - Pneumolysin potentiates oxidative inactivation of alpha-1-proteinase inhibitor by activated human neutrophils. AB - This study was designed to investigate the effects of the Streptococcus pneumoniae-derived, pro-inflammatory toxin, pneumolysin (8.37 and 41.75 ng/ml), on the oxidative inactivation of alpha-1-protease inhibitor (API) by chemoattractant-activated human neutrophils in vitro. The elastase inhibitory capacity (EIC) of API in supernatants from unstimulated neutrophils, neutrophils treated with pneumolysin only, or with the chemoattractant FMLP (1 microM) only, or the combination of the toxin with FMLP was measured by a colorimetric procedure based on the activity of added porcine elastase. The EIC of API was unaffected by exposure to pneumolysin only, unstimulated neutrophils, or neutrophils treated with pneumolysin only. However, exposure to FMLP-activated neutrophils resulted in a reduction of the EIC of API, which was significantly (P<0.05) augmented by pneumolysin (mean reductions of 16%, 43% and 83% for FMLP only and in combination with 8.37 and 41.75 ng/ml pneumolysin, respectively), and was attenuated by wortmannin (1 microM), an inhibitor of NADPH oxidase, the oxidant-scavenger methionine (100 microM), and depletion of Ca2+ from the cell suspending medium. These pro-proteolytic interactions of pneumolysin with chemoattractant-activated neutrophils may contribute to the invasiveness of the pneumococcus. PMID- 15338800 TI - Prospective evaluation of pneumonia severity index in hospitalised patients with community-acquired pneumonia. AB - The aim of the present study was to investigate whether the pneumonia severity index (PSI) could adequately predict the severity of community-acquired pneumonia (CAP) and could be used as a severity of illness classification system. Furthermore, reasons that may influence the decision to admit low risk patients were analysed. In a prospective study 260 patients with CAP were included. Stratification in five risk classes according to the PSI was compared with parameters that are closely related to severity of CAR A significant difference in severity parameters, such as length of stay (P < 0.001) and simplified acute physiologic score and acute physiologic and chronic health evaluation II score (P < 0.001) was found between the five risk classes. Furthermore, a positive British Thoracic Society (BTS) rule and modified BTS rule score was significantly more prevalent in the higher risk classes (P < 0.001). The patient population had an average 30-day mortality of 10% and a mean Intensive Care Unit (ICU) admission rate of 8%. The mortality rate and ICU admission rate significantly differed between the five risk classes (P < 0.001), in which the highest ICU admission rate (40.9%) and the highest mortality percentage (40.9%) were both found in risk class V. Several clinical factors (n = 64), such as an exacerbation of chronic obstructive pulmonary disease in 17 patients and clinical appearance of being ill in 16 patients, lack of improvement on outpatient antibiotic therapy (n = 15) and social circumstances (n = 3) were reasons that influenced the decision to hospitalise low risk patients (n = 82). The results show that the PSI adequately predicted the severity of CAP and can be used as a severity of illness classification in CAP. Clinical and social factors other than those mentioned in the PSI have to be considered when making the decision to hospitalise patients with CAP. PMID- 15338801 TI - Does respiratory syncytial virus subtype influences the severity of acute bronchiolitis in hospitalized infants? AB - Respiratory syncytial virus (RSV) subtypes A and B are present either simultaneously or alternate during yearly epidemics. It is still not clear whether clinical severity of acute bronchiolitis differs between the two subtypes. Reverse transcription polymerase chain reaction was used to subtype RSV in previously healthy infants hospitalized with RSV bronchiolitis during a winter epidemic. A severity index based on heart rate, respiratory rate, wheezing, difficulty in feeding and oxygen saturation was calculated upon admission. Infants infected with RSV subtype-A were found to have a significantly higher (more severe) clinical score than those infected with RSV-B. There was no statistically significant difference in duration of hospitalization or need of intensive care. Boys and infants younger than 3 months of age were also more severely affected than girls or older infants, respectively. These results support the notion that RSV-A-induced bronchiolitis is more severe than RSV-B induced one, in agreement with the majority of previously published studies. PMID- 15338802 TI - Resource use and risk factors in high-cost exacerbations of COPD. AB - Patients hospitalised for exacerbations contribute significantly to the total chronic obstructive pulmonary disease (COPD)-related healthcare costs. This study aimed to determine the resource use and costs of exacerbations by exacerbation severity and to identify risk factors for hospitalisation. Exacerbations and the details of all associated healthcare utilisation were recorded as part of a prospective cost-effectiveness analysis linked to two randomised controlled trials comparing tiotropium with ipratropium in 519 patients with stable COPD at study entry in the Netherlands and Belgium. Exacerbation-severity was rated by the physician. A Cox proportional hazards analysis was performed to identify independent risk factors of hospitalisation. Covariates that entered this analysis were smoking status, pack-years, body mass index, number of concomitant diseases, number of concomitant medications, use of inhaled steroids, physician visits prior to trial, FEV1% predicted, quality of life, baseline dyspnea index (BDI) and treatment arm. The mean number of exacerbations per patient was 0.70 (95%-CI:0.60, 0.81). About 10% of the exacerbations was severe, 47% moderate and 43% was mild. The mean costs of these exacerbations were Euro 4007 (95%-CI:2004, 6011), Euro 579 (390, 768) and Euro 86 (49, 124), respectively. In addition to treatment arm, a body mass index below 18.5 (RR:3.62), each additional concomitant diagnosis (RR:1.40) and a decrease of 1 point in the baseline dyspnea index (RR:1.18) were significant risk factors of hospitalisation. Exacerbations that were associated with a hospitalisation accounted for 90% of the total costs of exacerbations. Underweight, history of concomitant diseases and increased dyspnea (BDI score) are factors that are likely to identify patients who are at increased risk for generating high costs due to hospitalisation. PMID- 15338803 TI - Role of siroliumus, a novel immunosuppressive drug in heart and lung transplantation. AB - Lung and heart transplantation has become an accepted therapeutic option for patients with end-stage disease. However, the calcineurin-inhibitor-based immunosuppression often causes renal impairment. Therefore, sirolimus, a novel immunosuppressive agent, may serve as an alternative or complementary agent to calcineurin inhibitors. The aim of this review was to summarize the role of sirolimus in lung and heart transplantation. Although only a few, small studies have been conducted so far, the drug's mechanisms of action and low-toxicity profile make it a highly promising option. PMID- 15338804 TI - Variability of symptoms in mild persistent asthma: baseline data from the MIAMI study. AB - OBJECTIVE: To describe the variability of the asthma phenotype in patients with mild persistent asthma enrolled in the Mild Asthma Montelukast versus Inhaled Corticosteroid (MIAMI) study. METHODS: The variability of asthma rescue-free days, asthma symptoms, albuterol use, medical resource use, and exercise Limitations among patients with documented mild persistent asthma was compared between the month before study enrollment and the last 2 weeks of the run-in period. RESULTS: Patients eligible for randomization (n = 400), aged 15-85 years, exhibited symptoms (mean +/- SD) 3.6 +/- 1.3 days/week, beta-agonist use 3.5 +/- 1.3 days/week, and normal FEV1 (94.0 +/- 9.9% predicted) during the last 2 weeks of the run-in period. In the year before enrollment, medical intervention for asthma flares was common: 38.5% made office visits, 15.8% had oral corticosteroids, and 8.3% required emergency room or hospitalized care. In the month before enrollment, 11.8% experienced daily symptoms, and 28.3% had limitations of normal activity. Patients with daily symptoms in the month before study enrollment, compared with those having less-than-daily symptoms, experienced fewer rescue-free days (P = 0.024) and had more days per week with symptoms (P = 0.008) and requiring albuterol (P = 0.048) during the run-in; FEV1 was similar for both groups (93.1% vs. 94.2% predicted, respectively). CONCLUSION: Patients with mild persistent asthma reported a substantial disease burden in the year before enrollment. The asthma burden experienced by these patients both before and during the run-in period was of sufficient severity to support the recommendation that mild persistent asthma should be managed with daily controller therapy. PMID- 15338805 TI - Testosterone treatment improves body composition and sexual function in men with COPD, in a 6-month randomized controlled trial. AB - The aim of this study was to assess the effect of a low-dose testosterone on body composition and pulmonary function, as well as on quality of life, sexuality, and psychological symptoms in patients with chronic obstructive pulmonary disease (COPD). Twenty-nine men with moderate to severe COPD were allocated to receive either 250 mg of testosterone or placebo intra-muscularly, every fourth week, during the 26 weeks study period. Fat-free mass increased in the treatment group (P<0.05), and a significant difference between the treatment and the control group was seen after 26 weeks (P<0.05). Fat mass decreased in the treatment group (P<0.05), and there was a significant difference between the treatment and the control group after 12 weeks (P<0.01). A significantly better erectile function was reported in the treatment group at the final visit (P<0.05), and the overall sexual quality of life was significantly better in the treatment group after 12 weeks (P<0.05). No improvement in pulmonary function was found. In conclusion, administration of a low-dose testosterone to men with COPD for 26 weeks was associated with improvement of body composition, better erectile function and sexual quality of life. Furthermore, there were no clinical or biochemical side effects. PMID- 15338807 TI - [Dependence of elderly persons, long-term projections]. PMID- 15338811 TI - [Society facing advancing age]. PMID- 15338812 TI - [For a different social view on aging]. PMID- 15338813 TI - [Society's ambivalence towards elderly persons]. PMID- 15338814 TI - [Changing society's view of elderly persons]. PMID- 15338815 TI - [For a intergenerational society]. PMID- 15338816 TI - [Coping with incontinence and sexual disorders]. PMID- 15338817 TI - [Value of cognitive and psychosocial stimulation in the management of aged institutionalized patients with dementia]. PMID- 15338818 TI - [Nutritional assessment tools for malnutrition in the elderly]. PMID- 15338819 TI - [Occupational therapy between the city and the hospital]. PMID- 15338820 TI - [Art, culture and intergenerational relationships]. PMID- 15338821 TI - [Heat disorder protocol]. PMID- 15338822 TI - Quality managers: periodic performance review can be a powerful quality tool. Don't hesitate to put your processes under the microscope. PMID- 15338823 TI - Should you submit your PPR results to JCAHO? Take the opportunity to obtain valuable feedback. PMID- 15338824 TI - Key aspects of a recent Joint Commission survey. Surveyors focused on staff knowledge, patient care. PMID- 15338825 TI - Dramatically reduce your surgical infection rates. Set goals for 100% compliance. PMID- 15338826 TI - Worst practices used in conducting FMEA projects. Part 1 of a 2-part series. PMID- 15338827 TI - Other ways to check ESU analyzers. PMID- 15338828 TI - Research offers hope to treat apnea; and improve asset management. PMID- 15338829 TI - The pros and cons of outsourcing: which way should you go? PMID- 15338830 TI - From a clinical engineering perspective: understanding motivation and employee satisfaction. PMID- 15338831 TI - Possible causes of alarming... Line isolation monitors. PMID- 15338832 TI - Five tips to consider before buying refurbished equipment. PMID- 15338833 TI - Digital image storage. PMID- 15338834 TI - Smarter Web searching can yield better results. PMID- 15338835 TI - Calibration and maintenance: centralized system increases regulatory compliance. PMID- 15338836 TI - Attracting more women to the field... The few, the proud, the females. PMID- 15338837 TI - Safety in the health care facility. PMID- 15338838 TI - Altering the genetic code of management review. PMID- 15338839 TI - Use error: a nurse's perspective. PMID- 15338840 TI - Listserv use enhances quality and safety in multisite quality improvement efforts. AB - A listserv is an e-mail group to which people subscribe based upon common interests. We used a retrospective study to examine the relationship between listserv use and team success for health care quality improvement efforts. We hypothesized high listserv use would be associated with team success. Eighty seven Department of Veterans Affairs teams participated in facilitated quality improvement efforts to address three areas: improving safety in high-hazard areas, improving the disability evaluation process, and reducing falls and related injuries. We coded messages sent to the listserv according to sender (faculty or participant), team, and content. We correlated the volume of messages sent per team with team success and with team and facility characteristics. Teams with high listserv contributions were more likely to complete their first test of change, report facility use of nonpunitive methods of investigating medical incidents, and report their information systems were useful. We found a negative correlation between listserv contribution and the number of face-to-face meetings and a physician as an active team member, but we found no relationship between team success and listserv contribution. Team listserv contribution was not associated with team success in multisite quality improvement efforts. Successful teams may be accessing information on the listserv but not sending a message to indicate use. PMID- 15338841 TI - High-frequency chest compression: effect of the third generation compression waveform. AB - High-frequency chest compression (HFCC) therapy has become the prevailing form of airway clearance for patients with cystic fibrosis (CF) in the United States. The original square waveform was replaced in 1995 with a sine waveform without published evidence of an equality of effectiveness. The recent development of a triangle waveform for HFCC provided the opportunity to compare the functional and therapeutic effects of different waveforms. Clinical testing was done in patients at home with therapy times recorded with all sputum collected in preweighed sealable vials. The eight study patients with CF were regular users of a sine waveform device. They produced sputum consistently and were clinically stable. They used their optimum frequencies for therapy for each waveform and, for one week for each waveform, collected all sputum during their twice-daily timed HFCC therapies. After collection, these vials were reweighed, desiccated, and reweighed to calculate wet and dry weights of sputum per minute of therapy time. Frequency associated vest pressures transmitted to the mouth, and induced airflows at the mouth were measured in healthy volunteers. The pressure waveforms produced in the vest were, in shape, faithfully demonstrable at the mouth. In the healthy subject the transmission occurred in 2 ms and was attenuated to about 75% of the vest pressure for the triangle waveform and 60% for the sine waveform. All patients produced more sputum with the triangle waveform than with the sine waveform. The mean increase was 20%+ range of 4% to 41%. P value was <.001. Future studies of HFCC should investigate the other effects of the sine and triangle waveforms, as well as the neglected square waveform, on mucus clearance and determine the best frequencies for each waveform, disease, and patient. PMID- 15338842 TI - Planning for the future of technology management. PMID- 15338843 TI - Pharmacist-provided anticoagulation management in United States hospitals: death rates, length of stay, Medicare charges, bleeding complications, and transfusions. AB - We explored the associations between pharmacist-provided anticoagulation management in hospitalized Medicare patients and several major heath care outcomes: death rate, length of stay, Medicare charges, bleeding complications, and transfusions. Using the 1995 National Clinical Pharmacy Services database and the 1995 Medicare database for hospitals, data were retrieved for 717,396 Medicare patients treated in 955 hospitals for conditions requiring anticoagulant therapy. In hospitals without pharmacist-provided heparin management, death rates were 11.41% higher (chi2 (1) = 122.84, p<0.0001), length of stay was 10.05% higher (Mann-Whitney U test = 40039529342, p<0.0001), Medicare charges were 6.60% higher (U = 41004749266, p<0.0001), bleeding complications were 3.1% higher (chi2 (1) = 10.996, p=0.0009) and the transfusion rate for bleeding complications was 5.47% higher (chi2 (1) = 11.24, p=0.0008) than in hospitals with pharmacist provided heparin management. In hospitals without pharmacist-provided warfarin management, death rates were 6.20% higher (chi2 (1) = 19.20, p<0.0001), length of stay was 5.86% higher (U = 25730993838, p<0.0001), Medicare charges were 2.16% higher (U = 259955112970, p<0.0001), bleeding complications were 8.09% higher (chi2 (1) = 49.259, p<0.0001), and the transfusion rate for bleeding complications was 22.49% higher (chi2 (1) = 78.68, p<0.0001). Study hospitals without pharmacist-provided heparin management had 4664 more deaths, 494,855 more patient-days, 145 more patients with bleeding complications, and $651,274,844 more in patient charges; 9784 more units of whole blood were used in patients requiring transfusions for bleeding complications. Hospitals without pharmacist provided warfarin management had 2786 more deaths, 316,589 more patient-days, 429 more patients with bleeding complications, and $234,275,490 more in patient charges; 8991 more units of whole blood were used in patients requiring transfusions for bleeding complications. The implications of these findings are significant for the health care system, especially considering that the study population represents 28.25% of hospitalized Medicare patients who should receive anticoagulants, and that total Medicare admissions represent 35.02% of total admissions to United States hospitals. PMID- 15338844 TI - Pharmacoepidemiology I: a review of pharmacoepidemiologic study designs. AB - This article is the first of two parts on pharmacoepidemiology, a relatively new science that explores drug efficacy or toxicity using large observational study designs. The number of pharmacoepidemiologic studies published in medical journals has increased, as these studies have explored drug-related questions that at times cannot be answered in randomized clinical trials. Four pharmacoepidemiologic study designs will be discussed that explore the association between a specific pharmacologic agent and a disease of interest: cohort studies, case-control studies, case-crossover studies, and case-time control studies. PMID- 15338845 TI - Effect of tenofovir disoproxil fumarate on the pharmacokinetics and pharmacodynamics of total, R-, and S-methadone. AB - STUDY OBJECTIVE: To evaluate the potential effect of tenofovir disoproxil fumarate (DF) on the pharmacokinetics of methadone. DESIGN: Phase I, open-label, fixed-sequence, pharmacokinetic drug-drug interaction study. SETTING: Clinical research center. SUBJECTS: Fourteen volunteers receiving stable methadone maintenance therapy who were not infected with the human immunodeficiency virus. INTERVENTION: Tenofovir DF was added to the subjects' methadone regimens. MEASUREMENTS AND MAIN RESULTS: The pharmacokinetics of total, R-, and S-methadone were evaluated at baseline and after 2 weeks of daily tenofovir DF coadministration with a light meal. Steady-state tenofovir DF pharmacokinetics were evaluated at day 15. Bioequivalence testing was conducted of total, R-, and S-methadone area under the serum or plasma concentration-time curve during the 24 hour dosing interval at steady state (AUCss) and maximum concentration in serum or plasma (Cmax). Subjects were evaluated for changes in methadone pharmacodynamics by the Short Opiate Withdrawal Scale (SOWS) and pupillary diameter measurements at frequent intervals. Coadministration with tenofovir DF did not affect the pharmacokinetics of methadone. Geometric mean R-methadone systemic exposures, AUCss and Cmax, differed by 5% or less when methadone was dosed with tenofovir DF. Similar results were observed for S-methadone and for total methadone. Both AUCss and Cmax met the strict criteria for bioequivalence between the two study periods for total, R-, and S-methadone, indicating a lack of drug interaction when tenofovir DF was coadministered with methadone. No significant changes in SOWS scores or pupillary diameter measurements occurred, and no notable clinical adverse events were reported. CONCLUSION: Tenofovir DF pharmacokinetics were comparable to previously reported values of tenofovir DF in HIV-infected patients. Coadministration of methadone with tenofovir DF did not alter the pharmacokinetics or pharmacodynamics of total, R-, or S-methadone. Tenofovir DF may be given as part of a once-daily antiretroviral regimen in patients receiving methadone maintenance therapy. PMID- 15338846 TI - Impact of safety warnings on drug utilization: marketplace life span of cisapride and troglitazone. AB - STUDY OBJECTIVE: To evaluate the impact of safety alerts on the volume of cisapride and troglitazone usage. DESIGN: Retrospective database analysis. SETTING: University research center. MATERIAL: Idaho Medicaid claims data from January 1994--July 2000. MEASUREMENTS AND MAIN RESULTS: Monthly counts of total and new prescriptions filled for cisapride and troglitazone were analyzed graphically over time as a function of all prescriptions. New prescriptions were defined as those filled by patients who had not received the drug within the previous year. A binomial comparison of the 5 months before and after each safety alert was conducted by Poisson distribution. Overall and new cisapride usage increased after the first alert, which occurred in February 1995 (p<0.05). After the second alert, in September 1995, growth in new prescriptions ended but total prescriptions continued to grow (p<0.05). After the third alert, in June 1998, growth in total use ended and the number of new prescriptions declined (p<0.05). The final two alerts (June 1999 and January 2000) were met with significant declines (p<0.05 for both). Troglitazone was the subject of two alerts in October and December 1997. After these, overall usage increased (p<0.05), whereas the number o new prescriptions decreased (p<0.05). The third alert, in July 1998, caused no change as total prescription use continued to grow (p<0.05), whereas the number of new prescriptions decreased (p<0.05). A fourth alert, in June 1999, resulted in a decrease of overall usage and new prescriptions (p<0.05 for both). CONCLUSION: Numerous safety alerts were required for each drug before drug usage declined. The decline in overall use was slower than the decline in new prescriptions, possibly indicating a need for increased assessment of refilled prescriptions after the release of new safety data. PMID- 15338847 TI - Metformin monitoring and change in serum creatinine levels in patients undergoing radiologic procedures involving administration of intravenous contrast media. AB - STUDY OBJECTIVES: To evaluate the prevalence and magnitude of serum creatinine level elevations in patients receiving metformin who underwent radiologic procedures involving administration of intravenous contrast media, and to evaluate the efficacy of an electronic consultation in promoting timely evaluation of renal function after the procedure. DESIGN: Retrospective evaluation. SETTING: Veterans Affairs Medical Center. PATIENTS: Ninety-seven patients receiving metformin who underwent a radiologic procedure involving administration of intravenous contrast media over a 27-month period. MEASUREMENTS AND MAIN RESULTS: Ninety-seven patients underwent a total of 111 radiologic procedures with documented administration of intravenous contrast dye. Average time from procedure to laboratory follow-up, excluding one patient, was 2.62+/ 1.56 days. Average serum creatinine levels before and after the procedure were 1.10+/-0.19 and 1.13+/-0.23 mg/dl, respectively (p>0.05). Four patients developed contrast material-associated nephropathy. An additional four patients with borderline serum creatinine levels at baseline (1.4 mg/dl) had a serum creatinine level of 1.5 mg/dl or greater after the procedure. CONCLUSION: Our results indicated that electronic consultations result in timely evaluation of serum creatinine levels in patients receiving metformin who undergo a radiologic procedure involving intravenous contrast material. Also, the study suggests that nearly 4% of patients with diabetes mellitus and normal renal function may develop contrast material-associated nephropathy [corrected] with nonionic contrast material. In addition, about 8% of patients with diabetes treated with metformin (with baseline serum creatinine levels < 1.5 mg/dl) who undergo a procedure with nonionic intravenous contrast material acquire an increased risk (serum creatinine > or = 1.5 mg/dl) of lactic acidosis. These findings support the recommendations of the Food and Drug Administration regarding metformin monitoring in patients undergoing radiologic procedures involving administration of intravenous contrast media. PMID- 15338848 TI - Pharmacokinetics of modified oral calcitonin product in healthy volunteers. AB - STUDY OBJECTIVE: To assess the preliminary pharmacokinetics, pharmacodynamics, safety, and tolerability of single oral doses of a chemically modified salmon calcitonin product, CT-025, in healthy volunteers. DESIGN: Phase I, exploratory, five-way, open-label, nonrandomized, crossover study. SETTING: Clinical research center. SUBJECTS: Twelve healthy volunteers aged 22-44 years. INTERVENTION: A single oral dose of CT-025 was administered on 5 separate days with a 72-hour washout period between doses. Each dose consisted of CT-025 160 or 320 microg in varying relative concentrations of a mixture of excipients (formulations A, B, and C). MEASUREMENTS AND MAIN RESULTS: Serial blood samples were collected for determination of plasma CT-025, total serum calcium, and ionized serum calcium concentrations during the 4-hour period after dose administration. The data are the results from four of the five dosing days, when subjects received CT-025 in high and low concentrations of excipients. The data indicate that CT-025 was rapidly absorbed from the gastrointestinal tract. Mean peak plasma concentrations ranging from 51+/-51-110+/-59 pg/ml were observed 36-54 minutes after administration. Mean terminal elimination half-lives ranged from 54-76 minutes. There was a trend for the mean maximum concentration and area under the plasma concentration-time curve (AUC) from time zero to the time of the last quantifiable plasma concentration to increase with dose. Single oral doses of CT 025 160 and 320 microg were pharmacologically active, inducing a statistically significant decrease in total and ionized serum calcium concentrations. The rate of decrease in ionized serum calcium concentration was significantly related to the CT-025 dose. Single ora doses were well tolerated; some subjects experienced mild and transient nausea. CONCLUSION: Single doses of CT-025 were absorbed into the systemic circulation after oral administration and were well tolerated in healthy volunteers at doses up to 320 microg. These data suggest that oral delivery of salmon calcitonin becomes feasible with this product and support further clinical investigation of CT-025 as a treatment for osteoporosis and osteoporotic bone pain. PMID- 15338849 TI - Fixed-dose vasopressin compared with titrated dopamine and norepinephrine as initial vasopressor therapy for septic shock. AB - STUDY OBJECTIVE: To investigate the early blood pressure effects of vasopressin compared with titrated catecholamines as initial drug therapy in patients with septic shock. DESIGN: Retrospective cohort, single-center study. SETTING: Intensive care units at the Mayo Clinic, Rochester, Minnesota. PATIENTS: Fifty, 49, and 51 intensive care patients treated initially with vasopressin, norepinephrine, and dopamine, respectively. INTERVENTION: Patients received either intravenous infusion of fixed-dose vasopressin 0.04 U/minute or titrated infusions of norepinephrine or dopamine for low systemic arterial pressures. MEASUREMENTS AND MAIN RESULTS: Patients treated with vasopressin, norepinephrine, and dopamine were similar in all measured characteristics except for their score on the Acute Physiology and Chronic Health Evaluation (APACHE) III (dopamine > vasopressin, p=0.049), renal comorbidities (dopamine > vasopressin, p=0.03) and baseline mean arterial pressure (MAP) (norepinephrine < vasopressin, p=0.005 or dopamine < vasopressin, p=0.05). In all patients, MAP 1 hour before and 1 hour afte intervention, heart rate, and systolic blood pressure were obtained. No treatment differences were identified in achieving postvasopressin MAP after adjusting for APACHE III score, renal dysfunction, and baseline MAP. In patients receiving vasopressin, 28-day mortality was 52%, similar to those receiving norepinephrine (65%, p=0.28) and dopamine (60%, p=0.53). CONCLUSION: Initial, fixed-dose vasopressin infusions increased MAP to 70 mm Hg or greater at 1 hour in intensive care patients with septic shock, similar to titrated norepinephrine or dopamine. Fixed-dose vasopressin appears appropriate as an alternative agent for hemodynamic support in patients with septic shock. PMID- 15338850 TI - Economic analysis of intravenous plus oral amiodarone, atrial septal pacing, and both strategies to prevent atrial fibrillation after open heart surgery. AB - STUDY OBJECTIVES: To compare the cost-effectiveness of intravenous plus oral amiodarone, atrial septal pacing, and both strategies combined to prevent atrial fibrillation after open heart surgery. Secondary objectives were to compare the cost-effectiveness of amiodarone versus no amiodarone and of pacing versus no pacing, and to compare hospitalization costs of the various strategies. DESIGN: Piggyback cost analysis of a randomized, 2 x 2 factorial trial. SETTING: Urban academic hospital. PATIENTS: One hundred and sixty patients with coronary artery and/or valvular disease. INTERVENTION: Patients were randomized to receive amiodarone or matching placebo and then further randomized to receive atrial septal pacing or no pacing. MEASUREMENTS AND MAIN RESULTS: The economic analysis was conducted from a hospital perspective. Charges were converted to costs using cost:charge ratios. For the cost-effectiveness analysis, a joint distribution of costs and effectiveness was performed using the nonparametric bootstrap method. Amiodarone plus pacing significantly decreased the frequency of atrial fibrillation after open heart surgery, compared with amiodarone alone, pacing alone, and placebo. Total costs (mean+/-SD) were $27,026+/-30,226 for the placebo group, $22,725+/-17,661 for the amiodarone group, $33,868+/-60,309 for the pacing group, and $18,697+/-8174 for the amiodarone plus pacing group (p=0.27). In the joint distribution cost-effectiveness analysis, when compared with placebo, the probability of lower cost but higher effect (superiority) was 67% for amiodarone, 15% for pacing, and 97% for amiodarone plus pacing. In the multivariate analysis, preoperative beta-blockers and amiodarone were negatively associated with hospital costs (p<0.05). CONCLUSIONS: Data suggest that both amiodarone alone and the combination of amiodarone plus pacing are cost-effective compared with placebo. Additional comparative studies of these strategies are warranted to confirm these findings. PMID- 15338851 TI - Atomoxetine, a novel treatment for attention-deficit-hyperactivity disorder. AB - Atomoxetine is the first nonstimulant drug approved by the United States Food and Drug Administration (FDA) for the treatment of attention-deficit-hyperactivity disorder (ADHD), and the only agent approved by the FDA for the treatment of ADHD in adults. Atomoxetine is a norepinephrine transport inhibitor that acts almost exclusively on the noradrenergic pathway. Its mechanism of action in the control and maintenance of ADHD symptoms is thought to be through the highly specific presynaptic inhibition of norepinephrine. Clinical trials to evaluate the short term effects of atomoxetine in children and adults have shown that atomoxetine is effective in maintaining control of ADHD. Likewise, long-term trials have determined that atomoxetine is effective in preventing relapse of ADHD symptoms without an increase in adverse effects. A comparative trial of atomoxetine with methylphenidate in school-aged children indicated similar safety and efficacy without the abuse liability associated with some psychostimulants. The most commonly reported adverse effects in children and adolescents are dyspepsia, nausea, vomiting, decreased appetite, and weight loss. The rates of adverse events in the trials were similar for both the once- and twice-daily dosing regimens. The discontinuation rate was 3.5% in patients treated with atomoxetine versus 1.4% for placebo and appeared to be dose dependent, wit a higher percentage of discontinuation at dosages greater than 1.5 mg/kg/day. In clinical trials involving adults, the emergence of clinically significant or intolerable adverse events was low. The most common adverse events in adults were dry mouth, insomnia, nausea, decreased appetite, constipation, urinary retention or difficulties with micturition, erectile disturbance, dysmenorrhea, dizziness, and decreased libido. Sexual dysfunction occurred in approximately 2% of patients treated with atomoxetine. Atomoxetine should be used with caution in patients who have hypertension or any significant cardiovascular disorder. Overall, atomoxetine therapy in patient with ADHD appears to be effective in controlling symptoms and maintaining remission, with the advantages being comparable efficacy with that of methylphenidate, a favorable safety profile, and non-controlled substance status. Additional long-term studies are needed to determine its continued efficacy for those who require lifelong treatment, and comparative trials against other stimulant and nonstimulant agents. PMID- 15338852 TI - Role of clopidogrel in unstable angina and non-ST-segment elevation myocardial infarction: from literature and guidelines to practice. AB - Clinical trials are the backbone of treatment paradigm shifts and guideline development. In terms of acute coronary syndromes, the American College of Cardiology and the American Heart Association (ACC-AHA) have developed extensive guidelines to assist the practitioner in the appropriate use of drugs including antiischemic, anticoagulant, and antiplatelet agents. Clopidogrel, an adenosine 5'-diphosphate antagonist, is one such drug. Unfortunately, consensus guidelines are limited by the design of the clinical trials they reference. Clopidogrel trials have examined various outcomes in patients for a limited time frame, making longer term use of the drug difficult to justify. An ongoing study, estimated to be completed in 2005, is evaluating the long-term use of clopidogrel in high-risk patients. Aspirin, however, has become a lifelong therapy for many patients, based on clinical trials and medical experience. Patient-specific risk factors, the drugs' safety profiles, and costs, in addition to the ACC-AHA guidelines, must all be considered by clinicians when selecting the appropriate agent and its duration of use. PMID- 15338853 TI - The role of vasopressin in vasodilatory septic shock. AB - Septic shock that requires therapy with adrenergic agents is associated with high rates of mortality. Inappropriately normal or low serum concentrations of vasopressin contribute to the development of hypotension during sepsis. We critically evaluated the role of administering exogenous vasopressin to patients with septic shock. A computerized search of MEDLINE from January 1966--December 2003 and a manual search of relevant journals for abstracts were conducted. Eleven retrospective, six prospective cohort, and four prospective randomized studies were identified. Most studies evaluated short-term infusions of vasopressin at 0.08 U/minute or less as add-on therapy in patients requiring adrenergic agents. The results show that starting vasopressin in patients with septic shock increases systemic vascular resistance and arterial blood pressure, thus reducing the dosage requirements of adrenergic agents. These effects are rapid and sustained. Substantial enhancement of urine production, likely due to increased glomerular filtration rate, was shown in several studies. A few studies demonstrated clinically significant reduced cardiac output or cardiac index after vasopressin was begun, necessitating cautious use in patients with cardiac dysfunction. Vasopressin was associated with ischemia of the mesenteric mucosa, skin, and myocardium; elevated hepatic transaminase and bilirubin concentrations; hyponatremia; and thrombocytopenia. Limiting the dosage to 0.03 U/minut or less may minimize the development of these adverse effects. Vasopressin 0.03 U/minute or less should be considered if response to one or two adrenergic agents is inadequate or as a method to reduce the dosage of adrenergic agents. At present, vasopressin therapy should not be started as first-line therapy. Additional studies are needed to determine the optimum dosage, duration, and place in therapy of vasopressin relative to adrenergic agents. A multicenter, comparative study of vasopressin 0.03 U/minute as add-on therapy is under way and should provide mortality data. PMID- 15338854 TI - Angiotensin-converting enzyme inhibitors and stroke risk: benefit beyond blood pressure reduction? AB - Hypertension, a leading cause of morbidity and mortality, accounts for 25-49% of all strokes. Randomized placebo-controlled trials primarily with diuretics and beta-blockers administered in patients with hypertension have demonstrated a 38% reduction in primary stroke. Placebo-controlled trials with angiotensin converting enzyme (ACE) inhibitors have not been conducted in patients with hypertension. However, in a meta-analysis of four placebo-controlled trials of ACE inhibitors in patients with coronary heart disease and/or diabetes mellitus, the overall risk of primary stroke was significantly reduced. Results of the Heart Outcomes Prevention Evaluation trial, which produced a substantial reduction in stroke with an apparently small reduction in blood pressure, suggest that the benefit of ACE inhibitors may be related to their effects on the renin angiotensin-aldosterone system more than on blood pressure reduction. In active control comparisons in patients with hypertension, ACE inhibitors have demonstrated reductions in primary stroke risk similar to reductions with diuretics, beta-blockers, and calcium channel blockers. The data suggest that for primary prevention of stroke antihypertensive therapy should be individualized in patients. Relatively few data are available concerning the benefit of antihypertensive therapy in the secondary prevention of stroke. In patients who had experienced a stroke or transient ischemic attack, therapy with a diuretic or a combination of a diuretic plus an ACE inhibitor could be recommended based on available outcome studies conducted in this patient population. It is premature to conclude that the benefit of ACE inhibitor therapy in primary or secondary prevention of stroke is an effect independent of blood pressure reduction. Hypertension detection, treatment, and control in patients still must be the principal focus of clinicians for both primary and secondary prevention of stroke. PMID- 15338855 TI - Cost of a pharmacist-directed intervention to increase treatment of hypercholesterolemia. AB - STUDY OBJECTIVE: To evaluate the cost of a pharmacist-directed intervention that prompts physicians to treat hypercholesterolemia more aggressively in patients with coronary heart disease (CHD). METHODS: Health care resource use and CHD outcomes were evaluated for 612 patients with CHD followed for 2 years after an index hospitalization for an ischemic event. After discharge, the physicians of 309 patients who had been admitted from January 1--March 31, 1999, were contacted by telephone and mail concerning lipid profiles and statin therapy. These patients were the intervention group. Controls were 303 patients admitted from October 1--December 31, 1998; their physicians were not contacted. Costs of the physician-prompting intervention, clinic visits, laboratory tests, statin drugs, and CHD outcomes were compared between these two patient groups. RESULTS: The number of clinic visits, laboratory tests, and statins prescribed was significantly greater for the intervention group versus the controls. A significantly higher percentage of patients in the intervention group (55%) than in the control group (18%) achieved their National Cholesterol Education Program target low-density lipoprotein cholesterol level and had significantly better CHD outcomes. The cost of the physician-prompting intervention (pharmacist salaries, postage, telephone calls) was $102,941. For patients in the intervention and control groups, respectively, the cost of statin therapy was $352,365 and $200,087, the cost of clinic visits and laboratory tests $48,097 and $27,367, and the cost of coronary heart disease outcomes, such as myocardial infarction, coronary artery bypass graft, percutaneous transluminal and coronary angioplasty, $1,073,495 and $1,741,220. The total cost was $1,576,898 and $1,968,674, respectively, for patients in the intervention and control groups. Net savings was $1394/patient over the 2-year period. CONCLUSION: A relatively simple physician-prompting intervention involving patients with CHD significantly improved the use of lipid testing and statin therapy. Improved use of statins was associated with better CHD outcomes. As a result, the physician-prompting intervention was associated with cost savings. This intervention should be implemented for patients with CHD discharged after hospitalization for an ischemic event. PMID- 15338856 TI - Candida kefyr, an uncommon but emerging fungal pathogen: report of two cases. AB - Although Candida species are present as normal microflora of the human host, alterations in host defenses can lead to development of disease. Candida infections, ranging from urinary tract infections to bloodstream infections, are common in patients in the intensive care unit. Infections with non-albicans Candida sp are becoming more frequent, and resistance among these isolates is concerning. Candida kefyr is an uncommonly documented fungal pathogen. We report two cases of infection resulting from C. kefyr in our institution. The two patients had underlying disease states and drug therapies that increased the likelihood of developing an immunocompromised state. The C. kefyr isolates obtained from both patients were susceptible to amphotericin B, fluconazole, and itraconazole. Both patients had resolution of infection, one after receiving treatment with amphotericin B and the other with voriconazole. PMID- 15338857 TI - Fatal lactic acidosis associated with coadministration of didanosine and tenofovir disoproxil fumarate. AB - Lactic acidosis is an uncommon but potentially life-threatening adverse effect of didanosine. When given concomitantly with tenofovir disoproxil fumarate (DF), the area under the concentration-time curve of didanosine is increased by 48-60%. A 63-year-old man with human immunodeficiency virus (HIV) infection tolerated several didanosine-containing antiretroviral regimens. He developed generalized weakness, loss of appetite, weight loss, nausea, and vomiting 1.5 years after tenofovir DF was added to his didanosine-containing regimen. He was diagnosed with lactic acidosis and died after a 13-day hospital stay, when his lactate level increased to 189.7 mg/dl and his arterial blood gas pH value fell to 6.75. Health care providers should maintain a high index of suspicion for lactic acidosis in patients with HIV infection who receive didanosine and tenofovir DF concurrently. For patients receiving antiretroviral regimens containing this drug combination, it would be prudent to monitor lactate levels periodically. This is especially important when patients experience symptoms suggestive of lactic acidosis, such as weakness, abdominal pain, weight loss, nausea and vomiting, and shortness of breath. PMID- 15338858 TI - Elevated defibrillation threshold with venlafaxine therapy. AB - Depression is a common comorbidity in patients with heart failure or implantable cardioverter-defibrillators (ICDs). A 35-year-old woman with depression, nonischemic cardiomyopathy, and a history of cardiac arrest had an ICD implanted. During initial testing, the device failed to internally defibrillate the patient. Venlafaxine, an antidepressant with cardiac sodium channel blocking activity, was identified as a potential contributor to her elevated defibrillation threshold. After the venlafaxine was discontinued, the ICD was able to successfully internally defibrillate the patient. Clinicians should be aware of this potential adverse drug-device interaction. Further studies are needed to determine the clinical significance of venlafaxine therapy in patients with ICDs. PMID- 15338859 TI - [Microsurgery of temporal bone]. PMID- 15338860 TI - [Diagnosis and surgical management of petrous apex cholesteatoma]. AB - OBJECTIVE: To explore the etiology, diagnosis and surgical management of petrous apex cholesteatoma. METHODS: Twelve cases of petrous apex cholesteatoma (primary 3, secondary 9) were retrospectively studied. RESULTS: Primary petrous apex cholesteatoma was characterized by non otorrhea history, normal tympanic membrane appearance and the initially occurring symptoms of facial paralysis and hearing loss. In contrast, secondary petrous apex cholesteatoma was characterized by otitis media history, perforated drum, hearing loss and facial paralysis. The treatment principle for either primary or secondary cholesteatoma is to remove all the cholesteatoma. Based on the status of hearing and location and extent of cholesteatoma within temporal bone, 4 surgical approaches were taken in our study, which were translabyrinth, middle cranial fossa, combination of translabyrinth and middle cranial fossa, craniocervical combination approach (intralabyrinth approach). The complications of these surgical procedures and their management were discussed. According to result of 4 months to 15 years follow-up, there were no recurrence cases up to now, except one, which was operated 4 times as recurrence. Facial nerve anastomosis (3/12) or decompression (3/12) was performed simultaneously in the operations. The function of facial nerve partially recovered from V to IV of House and Brackmann grading in 2 out of 3 anastomosis cases and from IV to III in 2 out of 3 cases of decompression. CONCLUSION: Although there are some differences in symptoms and etiology between primary and secondary petrous apex cholesteatoma, complete surgical removal of lesion is necessary for both. Surgical approaches are decided according to location and extent of the lesion and hearing status. Our study indicated that open cavity operation had more advantage in terms of morbidity compared with the closed cavity operation, which closes the auditory canal as a blind sac. PMID- 15338861 TI - [Long-term results of small fenestra stapedectomy compared with large fenestra technique]. AB - OBJECTIVE: To evaluate the postoperative long-term hearing results of small fenestra stapedectomy versus large fenestra techniques. METHODS: A retrospective review of 102 ears of 94 patients who had stapedectomy (small fenestra technique) performed by the same surgeon between 1972-1990 was conducted. Patients undergoing stapedectomy and stapedotomy were followed for 6.4 +/- 2.2 and 6.6 +/- 3.0 month in initial postoperative and 14.5 +/- 3.0 and 7.4 +/- 2.2 years in late postoperative, respectively. RESULTS: There were no statistically significant differences in initial or late postoperative pure-tone (PTA), PTA air-bone gap, speech reception threshold (SRT) between the two groups. Ears treated by stapedotomy showed statistically better initial postoperative air-conduction threshold and initial 4-KHz air-bone gap closure, but the gap difference was not significant with late follow-up. The successful outcomes in both groups were stable over long-term follow-up. Results were the same when comparing the two procedures in the patients having undergone both. CONCLUSIONS: These results show that, in the operation of an experienced surgeon for otosclerosis, either large or small fenestra technique provides satisfactory and stable long-term results. PMID- 15338862 TI - [CT multiplan reconstruction images of disorder of stapes]. AB - OBJECTIVE: To evaluate the clinical application of multi-planar reformation (MPR) for the stapes with multi-slice spinal thin-section CT in the patients with disorder of stapes before prosthetic ossicular reconstruction and their impact on surgical decision. METHODS: Axial CT scanning of temporal bone was undergone in 50 volunteers. Multiplan reformatted images included coronal, sagittal and oblique MPR, were made. All the MPR images were compared with each other in order to show which one could reveal the whole structures of stapes better. The height of stapes was measured with the oblique MPR. CT findings were evaluated in 102 cases with disorder of stapes. Prosthetic ossicular reconstruction was made in 65 cases. RESULTS: The full stapes cannot be shown in axial, coronal and sagittal MPR images, however, it was shown in oblique MPR in all the cases. The highness of stapes was (3.3 +/- 0.4) mm in the abnormal group. There was chronic otitis media in 69 cases (90 ears) and congenital abnormalities of the stapes in 33 cases (47 ears) which included the dispart of incudostapedial joint in 5 cases (7 ears), defect of head and crus of stapes in 9 cases (13 ears), defect of one side of crus in 3 cases (4 ears) and absence of the oval window in 16 cases (23 ears). CONCLUSIONS: Coronal and oblique MPR images from Axial Spiral CT Data are essential for the pre-operative planning of prosthetic ossicular reconstruction. The ship, structure or defect of stapes can be shown in the oblique MPR images. It is important for the patient to choose the type of operation. PMID- 15338863 TI - [Round window area temporal bone section versus CT]. AB - OBJECTIVE: To locate round window area and its related structure on auris transection and CT for anatomical evidence of image diagnosis and clinical operation. METHODS: Fifteen normal head specimen fixed with 10% dehyde were scanned by high-resolution computed tomography on canthomeatal line. CT image (depth 1.00 mm, thick 1.00 mm) was obtained. Temporal bone-centered tissues were taken, decalcified, desiccated and socked with collodion, then embedded and made into sequential transactions (thick 1.00 mm). Lower surface of section was observed by both naked eyes and microscope, then scanned and photographed. Versus CT image, auditory ossicle, osseous semicircular canals, vestibule, round window, niche, cochlea, pyramidal eminence, internal acoustic meatus and cochlear aqueduct were identified respectively. RESULTS: There were 18-22 layers of temporal transection on CT image. Round window and round window niche always appeared on the 10th layer (R) and the 11th layer (L). The mean of the depth of anterior wall was 0.92 mm (R) and 0.90 mm (L), and depth 1.89 mm (R) and 2.04 mm (L). The average distance from niche to jugular fossa wall was 2.10 mm (R) and 2.39 mm (L). No significant difference among of thickness, depth and distance from niche to jugular fossa wall. CONCLUSIONS: Temporal bone transection specimen had a clear picture of anatomical position between round window area and its related structure. Versus CT, the result contributed to image diagnosis and operation on auris diseases. PMID- 15338864 TI - [Expression of aquaporin in rats' endolymphatic sac and kidney and the effect of anti-diuretic hormone on the expression of aquaporin]. AB - OBJECTIVE: To confirm the expression of aquaporin2,3,4 (AQP2,3,4, water channel protein) in rats' endolymphatic sac (ES) and kidney, and to investigate and compare the effects of antidiuretic hormone (AVP) and DDAVP [(deamino-Cys1,D Arg8)-Vasopressin, V2-receptor agonist] on the expression of AQP2 in rats' ES and kidney. METHODS: Thirty healthy Swards white rats were divided into the negative control, AVP group and dDAVP group, respectively, and were cardiaca perfused. The temporal bones and kidneys were taken out, then processed and sectioned by paraffin-embedded technique. The sections of ES were labeled with fluorescent antibody by immunohistochemical method, and kidney's with avidin-biotin peroxidase complex method (ABC). The expression of AQP-2,3,4 were confirmed in the ES of rats, and the different effects of the AVP and DDAVP onto the ES and kidney were observed. The slides used were analyzed by image-analyzer and the subsequent data were dealt with statistically. RESULTS: In the cytomembrane and cytoplasm of ES' epithelia, the constant and clear fluorescent reaction could be observed in normal control group with the first antibody of AQP2,3,4. Significant feeble fluorescent reaction of the first antibody of AQP-2 was revealed in AVP group and DDAVP group and showed much lower of gray (P < 0.01), less intensive of fluorescent (P < 0.01) under the fluorescence microscope. In the principal cell of renal collecting duct, it was on the contrary. Compared with the control group, significant stain was revealed in AVP group and DDAVP group and showed lower of gray (P < 0.05), greater density (P < 0.05), higher IOD (P < 0.05) and more stained area (P < 0.05). There is no different between the AVP group and DDAVP group in expression of AQP-2 in two sites. CONCLUSIONS: AQP-2,3,4 were expressed both in rats' epithelia of endolymphatic sac (ES) and principal cell of renal collecting duct. AVP promotes the expression of AQP2 in kidney but inhibits in ES. AVP maybe play important role to control the expression of AQP-2 in ES and kidney probably by the role of AVP-V2R-cAMP-AQP2. PMID- 15338865 TI - [Combination radiation and gene therapy for head and neck squamous cell carcinoma in the murine model]. AB - OBJECTIVE: To examine the antitumour efficacy and investigate immunological mechanism of combination therapy of IL-2 gene and IL-12 gene transfer with radiation in an immunocompetent murine model that parallel more closely the clinical therapy of head and neck squamous cell carcinoma (HNSCC). METHODS: Tumors were established in the floor of mouth in C3H/HeJ mice with SCCVII cell line. Lipid-DNA complexed (lipoplexes) by using polycationic liposome-Mediated transduction for HNSCC was transduced in tumor-bearing mouse by direct intratumoral gene transfer. The local tumor was radiated with a dose of 2 Gy in the second day. Tumor sizes were measured before and after the treatment as compared to the different single treatment groups and the controls. After tumors were subculture, the supernatants were collected for IL-2 and IL-12 expression by enzyme-linked immunosorbent assay (ELISA). Natural killer (NK) cell activity and cytotoxic T-lymphocyte (CTL) activity were also assayed by LDH method. CD4+ and CD8+ T-lymphocyte in tumor tissues were examined by immunohistochemistry. RESULTS: HNSCC tumor growth was significantly inhibited following combined IL-2 and IL-12 gene therapy with radiation as compared to IL-2 or IL-12 gene therapy with radiation, single IL-2 or IL-12 gene therapy, radiation alone and the controls. Increased secreted levels of IL-2 and IL-12 protein expression were found in combined and single IL-2 gene or IL-12 gene treated groups. The combination and single gene treated groups produced greater activation of CTL and NK than the controls of all concerned test. The significant CD4+ and CD8+ T lymphocyte infiltration was distributed and the numerous necrosis were seen in tumor tissues after combination therapy. CONCLUSIONS: Combined IL-2 gene and IL 12 gene therapy with radiation could significantly inhibited HNSCC tumor growth in the murine model and efficiently induced antitumor immunity of the host. PMID- 15338866 TI - [Reporter genes transfer into human squamous cell carcinoma of larynx factor receptor mediated non-viral GE7 polypeptide delivery system with a new epidermal growth]. AB - OBJECTIVE: To evaluate the efficiency of marker gene and reporter gene transfection into human squamous cell carcinoma of larynx implanted in nude mice with a new EGFR mediated non-viral polypeptide gene delivery system. METHODS: Labeled streptavidin biotin (LSAB) was applied to examine over-expression of epidermal growth factor receptor (EGFR) on the squamous cell carcinoma of larynx. The complex of marker gene or reporter gene and gene delivery system was prepared and subsequently inoculated with Hep 2 cells or injected into laryngeal carcinoma implanted on nude mice. RESULTS: Over-expression of EGFR was observed on inoculated Hep 2 cells and on 65% of laryngeal carcinoma specimen examined (15/23). Positive staining was located on the surface of the cells and some of them in the cytoplasma. Green fluorescence was observed on Hep 2 cells under fluorescent microscope 48 hrs after PEGFP-C1 gene transfection and became the strongest 72 hrs after transfection. The rate of transfection was 80% (78/97). Blue staining was noticed 12 hrs after beta-gal gene transfer and it became more remarkable 24 hrs after gene transfection with X-gal staining on the implanted human laryngeal carcinoma on nude mice. CONCLUSIONS: Squamous cell carcinoma of head and neck over-expressed EGFR and a non-viral GE7 polypeptide gene delivery system could transfer exogenous genes into laryngeal carcinoma cells and tissues with high efficiency and targeting both in vitro and in vivo suggesting that GE7 would be applicable as a new targeted, high efficient gene delivery system to the gene therapy of squamous cell carcinoma of head and neck. PMID- 15338867 TI - [Preliminary study on the functional localization of auditory cortex in the healthy young subjects using magnetic source imaging]. AB - OBJECTIVE: To evaluate the value of magnetic source imaging (MSI) in the functional localization of the primary auditory cortex. METHODS: The M100 waves of cortical auditory evoked magnetic fields (AEFs) evoked by 0.5, 1, 2, 4 and 8 kHz pure tones in 5 subjects and by 2 kHz pure tones in 25 healthy young subjects were measured respectively (16 males and 14 females, with the age from 20 to 32 years old) using a whole head 306 channel magnetoencephalography (MEG) system. The MSI obtained by superimposing functional MEG data on structural magnetic resonance image (MRI) was used to localize the M100 sources on the auditory cortex. RESULTS: The M100 waves of AEFs were clear and replicable in both hemispheres. The M100 sources were localized on the bilateral transverse temporal gyri in all 30 subjects. The localization of M100 on transverse temporal gyri varied with the changes of stimulus frequency. The localization of primary auditory cortex was asymmetrical between bilateral hemispheres, and the left hemisphere M100 dipoles were significantly posterior compared to the right M100 dipoles. The M100 responses appeared significantly earlier to the contralateral stimuli than that to the ipsilateral stimuli in both hemispheres. The dipole positions of M100 were independent of the side of the stimuli. CONCLUSIONS: The functional localization of the primary auditory cortex could be determined precisely by magnetic source imaging (MSI) with high spatiotemporal resolution. MSI would hold great promise as a noninvasive tool for the fundamental and clinical research in otology. PMID- 15338868 TI - [A study of the relationship between the expression of connexin43, E-cadherin and biological behaviors of human laryngeal cancer]. AB - OBJECTIVE: To investigate expression of connexin43 (Cx43) and E-cadherin (E-cad) and its relationship with the biological behavior in Laryngeal carcinomas. METHODS: Immunohistochemistry staining (PicTure two steps method) was used to detect connexin 43 and E-cadherin expression in 60 paraffin-embedded laryngeal carcinomas and 10 normal epithelia around tumors. All the patients were followed up more than five years. RESULTS: Cx43 and E-cad were expressed by normal epithelia with typical membranous staining 10 normal epithelia strongly expressed Cx43 and E-cad. The reduction rate of Cx43 expression in laryngeal carcinomas was 30.0%. The significant relationship was observed between low Cx43 expression and differentiation, lymph node metastasis and recurrence (chi2 = 15.09, 6.41, 3.86, P < 0.01, 0.05, 0.05), but there was no significance between low Cx43 expression and prognosis (chi2 = 2.65, P > 0.05). 41.7% laryngeal carcinoma showed reduced or no expression of E-cad. Tumor cell dedifferentiation correlated with reduced expression for E-cad (chi2 = 15.07, P < 0.01). Absent or low E-cad expression was observed more frequently in patients with local recurrence and lymph node metastasis and with less than 5-year survival period (chi2 = 5.35, 6.65, 5.14; P < 0.05, 0.01, 0.05). There was a positive relationship between the expression of Cx43 and E-cad in same sample (r = 0.63, P < 0.0001). CONCLUSIONS: Laryngeal cancer presented inactivation of Cx43 gene and E-cad gene and down regulation of Cx43 and E-cad proteins. The level of Cx43 and E-cad may be a sensitive predictor of differentiation, invasion, lymph node metastasis and recurrence. E-cad is also a prognostic factor for patients with laryngeal cancer. PMID- 15338869 TI - [Clinical evaluation of 18-fluorine fluorodeoxyglucose positron emission tomography-CT imaging in head and neck tumors]. AB - OBJECTIVE: To study the clinical value of 18-fluorine fluorodeoxyglucose (18 F FDG) positron emission tomography (PET)-CT imaging in diagnosis and detection of primary tumors and suspicious recurrence in head and neck cancers. METHODS: Twenty-seven patients with primary tumors or metastases in head and neck underwent 18F-FDG PET-CT imaging, and compared with clinical data. RESULTS: 1. In 4 NPC diagnosed pre-radiation patients the 18F-FDG PET-CT imaging in nasopharynx was all positive, 2 of them were detected with neck lymph node metastases, which were confirmed later histopathologically. In 12 NPC cases after radiation, 4 were not found any evidence of tumor with PET-CT imaging and clinically, 4 were detected neck recurrence and confirmed histopathologically, 2 had remote metastases confirmed by other imaging, 2 were detected and confirmed recurrence in primary sites and one of them in regional and remote site. 2. In 4 neck metastatic patients with unknown primary tumor, the primary tumors were all identified, 3 of them were confirmed histopathologically. 3. Seven patients with other head and neck tumors after therapy underwent 18F-FDG PET-CT imaging. Two patients had no evidence of recurrence. Five patients were detected recurrence or metastases, 4 of them were confirmed histopathologically, one of them refused biopsy. CONCLUSIONS: With metabolic image of PET and anatomic image of CT, 18F FDG PET-CT is a kind of very useful way in diagnosing primary tumor and detecting the recurrence and metastases in patients with head and neck tumor. PMID- 15338870 TI - [5-HT2C receptor subtype modulate production of secretory beta-amyloid protein precursor in incubated rat hippocampal slices]. AB - AIM: To determine whether serotonin, a major neurotransmitter in brain, can modulate the production of secretory beta-amyloid protein precursor (sAPP) by activation of serotonin 5-HT2C receptor. METHODS: The hippocampal slices of rats were incubated with various concentrations of serotonin, M-110, or L-107. sAPP released into the incubation medium were assayed by Western blot analysis assay with monoclonal antibody 22C11 for 2 h. RESULTS: Various concentrations of serotonin (1.0 x 10(-2) - 1.0 x 10(3) micromol x L(-1)), M-110, a serotonin 5 HT2C agonist (1.5 x 10(-6) - 1.5 x 10(3) micromol x L(-1)), showed positive effect on the production of sAPP while L-107, a serotonin 5-HT2C antagonist (1.0 x 10(-9) - 1.0 x 10(3) micromol x L(-1)), showed negative effect on the production of sAPP over controls. CONCLUSION: Serotonin modulates production of secretory amyloid beta-protein precursor through serotonin 5-HT2C receptor in incubated rat hippocampal slices. PMID- 15338871 TI - [Protective effect and mechanism of pharmacologic preconditioning induced by sodium ferulate on primary cultured myocardial cell injury by anoxia/reoxygenation]. AB - AIM: To study the preconditioning effects and mechanism of action of sodium ferulate (SF) on primary cultured myocardial cell injury induced by anoxia/reoxygenation. METHODS: Cultured myocardial cells of neonatal SD rats were randomly divided into ten groups: control group: without any treatment; anoxia/reoxygenation group (A/R), reoxygenation of 1 h following anoxia of 3 h; anoxia preconditioning group (AP), reoxygenation of 30 mins following anoxia of 30 mins, three times before the same procedure as group A/R; SF preconditioning groups, 20 mins of SF (1.68, 0.42, 0.105 mmol x L(-1) in final concentration) preconditioning followed by 10 mins wash out before A/R; K+ATP channel blocker group, NOS inhibitor group and PKC inhibitor group, adding gliberclamide, L-NAME, ploymyxin B at final concentration of 12 g x mL(-1), 50 micromol x L(-1), 50 micromol x L(-1), to culture medium respectively 10 min before the same procedures as SF preconditioning group (1.68 mmol x L(-1)). Myocardial cells pulse rate and rhythm, myocyte viability, the activity of LDH and CK in culture, the contents of intracelluar MDA, LD in myocardial cells, the activity of SOD and GSH-Px of the cultured myocardial cell were measured at the end of experiment. RESULTS: Compared with control group, anoxia/reoxygenation caused great increases of levels of LDH, CK, MDA and LD (P < 0.01), decreases of myocardial cells pulse rate, cell viability, SOD and GSH-Px (P < 0.01); SF preconditioning significantly attenualed these increases and decreases. Glib, L-NAME, and Ploy B partly abolished the effects of SF preconditioning. CONCLUSION: SF preconditioning is effective in protecting myocardial cells from anoxia injury. The cardioprotective effect of SF preconditioning is produced by multiple factors. PMID- 15338872 TI - [The ion targets of arrhythmias induced by ouabain and aconitine in guinea pig and rat ventricular myocytes]. AB - AIM: To observe the effects of ouabain and aconitine on APD and ion channels in isolated guinea pig and rat ventricular myocytes; to elucidate the action mechanisms of these two drugs and set up new arrhythmic models on cellular level. METHODS: In isolated ventricular myocytes of guinea pig and rat, the effects of ouabain and aconitine on APD, ICa-L, Ik, Ito and Ik1 were observed using the whole cell patch clamp technique. RESULTS: Ouabain (5 micromol x L(-1)) obviously prolonged the APD90, increased ICa-L, decreased Ik and Ik1 in guinea pig ventricular myocytes. Aconitine (1 micromol x L(-1)) lengthened the APD90, increased ICa-L, decreased Ito and increased Ik1 in rat ventricular myocytes. CONCLUSION: The targets on ouabain- and aconitine-induced arrhythmias included APD, ICa-L, Ik, Ito, and Ik1. APD, ICaL, Ik and Ito must be the powerful ones, both in arrhythmic and antiarrhythmic courses. The ouabain- and aconitine- induced arrhythmic models on cellular level were built to study the antiarrhythmic mechanisms of chemicals and evaluate new drugs. These two new-type models in vitro were stable, liable, repeatable and economic, which were superior to those typical models in vivo. PMID- 15338873 TI - [Reversal of multidrug resistance by lomerizine in K562/ADM cells]. AB - AIM: To study the effect of lomerizine (Lom) on the reversal of multidrug resistance (MDR) in K562/ADM cells and its mechanism. METHODS: MTT assay was used to determine the influence of Lom on the cytotoxicity of adriamycin (ADM). The effect of Lom on the apoptosis induced by ADM and vincristine (VCR) in K562/ADM cells was detected using flow cytometry. Intracellular accumulation of ADM was measured by fluorescence spectrophotometry. Flow cytometry was used to investigate the efflux of rhodamine 123 (Rh123) and the expression of P glycoprotein (P-gp) in K562/ADM cells. RESULTS: Lom increased the cytotoxicity of ADM and the apoptosis induced by ADM or VCR in K562/ADM cells. At the concentration of 3, 10 and 30 micromol x L(-1), Lom reduced the IC50 value of ADM from 79.03 micromol x L(-1) to 28.14, 8.16 and 3.16 micromol x L(-1), respectively. Lom increased the intracellular accumulation of ADM and inhibited the efflux of Rh123 in K562/ ADM cells. No change in P-gp expression was observed after the treatment of Lom for 72 h. CONCLUSION: Lom had strong reversal effect on MDR in K562/ADM cells by inhibiting P-gp function. PMID- 15338874 TI - [Relationship between M3 receptor and myocyte apoptosis induced by acute myocardial infarction]. AB - AIM: To explore the effects of M3 receptor on myocyte apoptosis induced by acute myocardial infarction in rats. METHODS: Rat model was induced by ligation of the anterior branch of the left coronary artery. All animals were divided into four groups: sham-operated group, occlusion group, choline group (10 mg x kg(-1), iv), and 4DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) group (0.12 mg x kg( 1), iv). The serum malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined. The infarct size areas on the myocardium were identified by TTC staining. The apoptosis in cardiomyocyte was detected by TUNEL assay and apoptosis-related proteins in Bcl-2 and Fas expression were measured by immunohistochemistry assay. RESULTS: M3 receptor agonist choline reduced serum MDA content and increased SOD activity. The myocardial expression of Bcl-2 was increased, whereas the expression of Fas was decreased by choline. However, blockade of M3 receptor by 4DAMP completely inhibited these effects of choline on cardiac myocytes. CONCLUSION: Activation of M3 receptor has protective effect on myocyte apoptosis induced by acute myocardial infarction in rat, and this effect might be related to modulating the expression of some immediateearly genes including Bcl-2 and Fas. PMID- 15338875 TI - [Synthesis and antitumour activities of some pteridine derivatives]. AB - AIM: To study the synthesis and antitumour activities of some aryl-substituted pteridines. METHODS: A series of aryl-substituted pteridines were synthesized from 4, 6-diamino-5-nitrosopyrimidines by cyclization with 4 aminophenylacetonitriles. The antitumour activities were tested by MTT method. RESULTS: Nine new compounds (I-III) were synthesized and their structures were characterized by EA, IR, 1HNMR and MS spectra. Compounds I-III showed antitumour activities in vitro. CONCLUSION: Compounds I-III showed remarkable antitumour activities in vitro. No interaction was determined between the title compounds and calf thymus DNA. It indicated that these compounds possibly inhibit dihydrofolate reductase (DHFR) or other enzymes on which folic acid depends. PMID- 15338876 TI - [A new steroidal alkaloid from the bulbus of Fritillaria wabuensia]. AB - AIM: To investigate the chemical constituents of the bulbs of Fritillaria wabuensia. METHODS: Chromatography techniques were used to isolate the chemical constituents. EI-MS, 1HNMR, 13 CNMR and DEPT were used to determine the structures of the isolated constituents. RESULTS: Three alkaloids were isolated from the bulbs of Fritillaria wabuensia, and were identified as imperialine (I), imperialine-beta-N-oxside (II), isoverticine-beta-N-oxide (III). CONCLUSION: Isoverticine-beta-N-oxide was isolated from the bulbs of Fritillaria wabuensia for the first time. Isoverticine-beta-N-oxide is a new alkaloid. PMID- 15338877 TI - [Water-soluble chemical constituents of Swertia davidi Franch]. AB - AIM: To study the active constituents of Swertia davidi Franch. METHODS: Column chromatographies on silica gel, Sephadex LH-20 and Diaion-201 et al. were used to isolate and purify the chemical components. Their structures were identified by UV, IR, MS, NMR and 2D-NMR. RESULTS: These compounds were elucidated as 8-O-beta D-glucopyranosyl-1, 3, 5-trihydroxyxanthone (I), 5-O-beta-D-glucopyranosyl-1, 8 dihydroxy-3-methoxyxanthone (II), 5-O-beta-D-glucopyranosyl-1, 3, 8 trihydroxyxanthone (III) and swertamarin (IV). CONCLUSION: Compound III is a new xanthone glucoside. The other compounds were isolated from this plant for the first time. PMID- 15338878 TI - Structures of four new triterpenoid saponins from the leaves of Oplopanax elatus Nakai. AB - AIM: Isolation and structural elucidation of the triterpenoid saponins of Oplopanax elatus Nakai. METHODS: Solvent extraction and column chromatography were used to isolate the triterpenoid saponins, physico-chemical constants and spectroscopic analysis were employed for structural elucidation. RESULTS: Four newtriterpenoid saponins named cirenshenoside S (1), cirenshenoside T (2), cirenshenoside U (3) and cirenshenoside V (4) were isolated, and their structures were elucidated to be 3-O-beta-D-glucopyranosyl 3beta,23-dihydroxylup-20 (29)-en 28-oic acid 28-O-alpha-L-rhamnopyranosyl (1 --> 4)-beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranoside (1), 3-O-beta-D-glucopyranosyl hederagenin 28-O-alpha-L rhamnopyranosyl (1 --> 4)-beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranoside (2), 3-O-beta-D-glucopyranosyl 3beta-hydroxyolean-9(11),12-dien-28-oic acid 28-O alpha-L-rhamnopyranosyl (1 --> 4)-beta-D-glucopyranosyl (1 --> 6)-beta-D glucopyranoside (3) and 3alpha-hydroxyolean-12-dien-23,28-dioic acid 28-O-alpha-L rhamnopyranosyl (1 --> 4 )-beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranoside (4), respectively. CONCLUSION: Compounds 1-4 are new triterpenoid saponins and isolated from the leaves of Oplopanax elatus Nakai for the first time. PMID- 15338879 TI - [Study on methods for quality control of recombinant human neu epitope peptide 12]. AB - AIM: To establish methods and requirements for the quality control of recombinant human neu epitope peptide 12. METHODS: Biological activity of recombinant human neu epitope peptide 12 was evaluated in FVB/N transgenic mice (TgN MMTV neu 202 Mul, Jackson Lab., USA) administered with samples. The percentage of antibody positive mice detected by ELISA was used in the biological activity evaluation. The peptide map was performed by peptic digestion. The antigen content was determined by SEC-HPLC. RESULTS AND CONCLUSION: The quality control methods, such as biological activity, peptide map, antigen content, and the requirements for the quality control of recombinant human neu epitope peptide 12 were established. The established methods and requirements were already used for the quality control of recombinant human neu epitope peptide 12 products. PMID- 15338880 TI - [Separation and identification of alkaloids in Stephania tetrandra by capillary electrophoresis electrospray-MS/MS]. AB - AIM: To establish a method for separation and identification of alkaloids in Stephania tetrandra S. Moore methanol extracts by using non-aqueous capillary electrophoresis interfaced with electrospray ionization ion trap mass spectrometry. METHODS: The molecular ions or adducts of alkaloids and fragments of specific parent ions were used for the identification. An uncoated capillary (86 cm x 75 microm ID, on-line UV detection occurred at 21 cm from the inlet of the capillary) was used. Ammonium acetate (50 mmol x L(-1)) containing 4% HAc in methanol was used as the running buffer; separation voltage was 25 kV. A coaxial sheath flow interface was used as the CE-MS interface; the electrospray voltage was 4.5 kV; the temperature of aluminium capillary was 170 degrees C; 60% isopropanol-39% water-1% HAc was used as the sheath liquid with the flow rate of 5 microL x min(-1); the collision energy of MS-MS was set at 30% and the least ion counts was 1 x 10(5). RESULTS AND CONCLUSION: The alkaloids in Stephania tetrandra S. Moore methanol extracts were separated and identified by CE-ESI MS/MS. The proposed method is of high accuracy and can be used for the investigation of traditional Chinese medicine. PMID- 15338881 TI - [Determination of clonidine in rabbit plasma by HPLC-MS]. AB - AIM: To determine clonidine in rabbit plasma by LC-MS. METHODS: The LC-MS system consisted of Waters Alliance 2790 HPLC and Micromass ZQ-4000 MS. The HPLC was performed by using XTerra C18 (150 mm x 2.1 mm ID, 5 microm). The mobile phase, consisting of acetonitrile/ammonium hydrogen carbonate solution, was maintained to a flow-rate of 0.2 mL x min(-1) and the linear gradient elution was adopted. Mass spectrum was obtained by using electrospray ionization interface and the m/z of SIM was 230. RESULTS: The average recovery was high and the method was reproducible. The calibration curve showed good linearity in the range of 1 - 80 microg x L(-1), the lowest limit of detection was 0.05 microg x L(-1). The Cmax, AUC0-t, and Tmax value of the pharmacokinetics parameter were (27 +/- 9) microg x L(-1), (5,352 +/- 1,121) microg x L(-1), (79 +/- 17) h. CONCLUSION: The results demonstrated that the method had high sensitivity, good selectivity, accuracy and precision. It is used to determine the clonidine concentration in plasma. The transdermal patch can deliver clonidine to the surface of rabbit skin stably for periods of up to 1 week after a single application. PMID- 15338882 TI - [Studies on chiral inversion of dextropantoprazole in human]. AB - AIM: To study the chiral inversion of dextropantoprazole in human. METHODS: Three healthy Chinese male volunteers after an oral dose of 40 mg dextropantoprazole. An HPLC method was developed and used to determine the total plasma concentrations of each enantiomer. The ratios of the enantiomers in plasma samples were measured on a Chiral-AGP column. The plasma concentration of each enantiomer was then calculated using the ratios of enantiomers and total concentrations of the two enantiomers previously measured. RESULTS: The AUC0-t of levopantoprazole was only 1.5% of the total AUC0-t of enantiomers. CONCLUSION: The chiral inversion from dextropantoprazole to levopantoprazole does not occur in the three healthy Chinese male volunteers after an oral dose of 40 mg dextropantoprazol. PMID- 15338883 TI - [Selective tumor-accumulation of HPMA copolymer-mitoxantrone conjugates]. AB - AIM: To increase the accumulation of mitoxantrone in solid tumor by synthesis and characterization of N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer mitoxantrone conjugate (p-DHAQ). METHODS: HPMA copolymer-mitoxantrone conjugate was prepared by free radical precipitation copolymerization method. The in vitro stability of conjugate was investigated under different conditions. Biodistribution was examined in mice bearing Ehrlich solid tumor. RESULTS: The p DHAQ conjugate was characterized by UV, HPLC and size exclusion chromatography. The conjugate was stable in buffers of different pH and in mice plasma while the rate of drug liberation was faster in tumor. It appeared that the circulation lifetime of HPMA copolymer-bound mitoxantrone were three times more than that of the drug in free form. The AUC of p-DHAQ was three times more than the AUC of free drug. The p-DHAQ level in heart was five times lower than free drug. This reduces the possibility of toxicity to the heart. CONCLUSION: HPMA copolymer mitoxantrone conjugate was successfully synthesized and characterized. The biodistribution results showed the possibility of targeting anticancer drug mitoxantrone with secondary amino residue to the tumor tissue by HPMA copolymer as carrier. PMID- 15338884 TI - [The enhancing effect of tomato lectin modified liposomes of insulin on oral absorption in mice]. AB - AIM: To investigate the enhancing effect on insulin absorption through GI tract in mice by using tomato lectin (TL) modified liposomes as the carrier. METHODS: TL-phosphatidylethanolamine (PE) conjugate (TL-PE) was synthesized by using carbodiimide cross-linking method, then the compound was incorporated into the conventional liposomes of insulin. The agglutination test was performed to examine TL biological activities after synthesis and incorporation. When TL modified liposomes were administrated orally to the normal mice or diabetic mice at insulin dose of 350 u x kg(-1), the hypoglycemic effect was determinated according to the blood glucose level. RESULTS: The blood glucose levels of the normal mice were reduced by modified liposomes. The glucose levels were (85 +/- 5)% at 4 h, (54 +/- 11)% at 8 h, (57 +/- 6)% at 12 h postdose compared with the glucose levels prior to oral administration respectively. However, the conventional liposomes and saline have no hypoglycemic effect. The blood glucose levels of the diabetic mice were obviously reduced by TL modified liposomes, the glucose levels were (38 +/- 13)% at 4 h, (50 +/- 15)% at 8 h, (50 +/- 16)% at 12 h respectively. CONCLUSION: TL modified liposomes promote the oral absorption of insulin due to the specific-site combination on GI cell membrane. PMID- 15338885 TI - [Research of ginsenosides in kou zi qi using HPLC-MS-MS]. AB - AIM: To analyze the ginsenosides in Kou zi qi (rhizomes of Panax japonicus C. A. Mey. var. major (Burkill) C. Y. Wu et K. M. Feng), and to supply evidences for chemotaxanology of Panax species and clinical uses of Kou zi qi. METHODS: The ginsenosides were isolated by HPLC, then the positive- and negative-ion API-MS/MS of constituents collected from HPLC were measured. RESULTS: Eight ginsenosides were identified as ginsenoside Re, ginsenoside Ro, chikuseksusaponins IV, IVa, notoginsenoside R2, ginsenosides Rb1, Rc and Rd, respectively, based on comparison of retention time with those of standards by HPLC, and analysis on their API-MS/MS data. Ginsenoside Ro and chikuseksusaponin IVa are the major components of Kou zi qi. CONCLUSION: This plant had a close relationship to P. stipuleanatus, P. zinginensis and P. japonicus var major; a relatively remote relationship to P. ginseng and P. quinquefolius, in a view of chemotaxanology. Ginsenoside Ro and chikuseksusaponin IVa might be the anti-inflammatory constituents of Kou zi qi. PMID- 15338886 TI - [Description of the release of sodium ferulate from hydroxypropyl methylcellulose based matrix tablets in vitro]. AB - AIM: To elucidate the mechanism and to present suitable models for the release of sodium ferulate (SF) from hydroxypropyl methylcellulose (HPMC) based matrix tablets. METHODS: The characteristics and mechanisms of drug release were analyzed from the point of thermodynamic, swelling and diffusion effect. Despite the classical equation of Higuchi, the semi-empirical power law and quadratic curve were also adopted to analogize the release of SF from HPMC based tablets in vitro. RESULTS: The first release stage of SF was mainly controlled by Fick diffusion, and the rest SF released mainly according to case II transport process caused by the swelling effect of HPMC. The decreased Tg of HPMC, resulted from the entered water, enhanced the release of SF. The power law was possible for the first released 60% SF, but unfit for the last 40% SF. The quadratic curve expressed equation can illuminate the release of SF. CONCLUSION: For the HPMC system, drug release undertakes the Fick diffusion process followed by the extend of polymer chain. The nonlinear quadratic equation might be valuable to explain the entire drug release from HPMC-based delivery system. PMID- 15338887 TI - [Transport of proteins and peptides across human cultured alveolar A549 cell monolayers]. AB - AIM: An in vitro cultured monolayer system of alveolar epithelial cells was used as a model to investigate the transport pathway peptides or proteins, salmon calcitonin (sCT), insulin (INS), recombinant hirudin (rHAV2), and recombinant human growth hormone (rhGH), in pulmonary epithelium in vivo. METHODS: Human lung adenocarcinoma A549 cells formed continuous monolayers with growing polycarbonate filters of Transwell plate. Transport studies of macromolecules in the monolayer system were carried out after 6 days in culture. The transport of peptides or proteins with MW 3,400 - 22,000 was studied in cultured human lung adenocarcinoma A549 cell monolayers at different conditions. RESULTS: The results showed that the apparent permeability coefficients (Papp) of these macromolecules across A549 cell monolayers ranged from 2 x 10(-6) to 5 x 10(-6) cm x s(-1) and exhibited good inverse correlation with molecule weight. No concentration, direction and temperature dependence were observed in the permeation of sCT, INS and rHAV2. While the Papp of rhGH in the BA direction (2.25 x 10(-6) cm x s(-1)) was significantly less than that in the reverse direction. The Papp values of rhGH were concentration and temperature independent in the AB direction. CONCLUSION: These findings suggest that the hydrophilic peptides and proteins, salmon calcitonin, insulin, recombinant hirudin, and recombinant human growth hormone used in this study, appeared to penetrate the A549 cell monolayers via a paracellular pathway by passive diffusion mechanism. PMID- 15338888 TI - [Homocamptothecins--novel promising anticancer drugs as inhibitors of topoisomerase I]. PMID- 15338889 TI - [Analyzing the epidemic situation due to tuberculosis and ways of optimization of its control in the penitentiary facilities of the Ulyanovsk Region]. AB - The dynamics of epidemic parameters and the specific features of development of tuberculosis were analyzed in prisoners in 1989-2001. It was ascertained that 98.7% of them were infected with tuberculosis and most of them belonged to a tuberculosis high-risk group. In 67.5% of those who had fallen ill, the process was detected within the first 3 years of their confinement. A scheme of interaction of main propulsive forces of an epidemic process in tuberculosis (exogenous and endogenous infections and risk factors) is presented. It has been found that surveys for tuberculosis are not a deterrent for a rise in the incidence rates due to endogenous infection. The use of factorial analysis and systems approach in the epidemiology of tuberculosis has explained some regularities in the development of an epidemic process, revealed reasons for the aggravated tuberculosis situation in penitentiary institutions, and considered prospects for its improvement. PMID- 15338890 TI - [Impact of patients with tuberculosis in the penitentiary system on the infection and morbidity rates of the population]. AB - The paper shows infection and morbidity rates in children in the flat and territorial foci of bacterial isolates released from confinement institutions and in the socially favorable foci, as well as infection rates in children in the areas when correctional facilities are located. Patients with tuberculosis who are in the correctional facilities have been found to have a negative impact on the infection rates in children living in the areas when correctional facilities are located. So have those released from penitentiary facilities on the infection and morbidity rates in children in the flat and territorial foci. PMID- 15338891 TI - [Sociomedical factors influencing mortality in patients with tuberculosis]. AB - The paper presents a clinical and social characterization of patients (n = 747) who died from tuberculosis in Volgograd during 2000-2002 on the basis of an analysis of all fatal cases. The sociomedical factors influencing the mortality of patients with tuberculosis have been determined and the sociomedical portrait of a patient with tuberculosis has been created. PMID- 15338892 TI - [Infection of children with tuberculosis and the nature of tuberculin sensitivity in the infected]. AB - In the Kharkov Region, the rate of tuberculosis infection in children was 28.8%; among them children with a curve of tuberculin reactions was 3.8%; most of them had a moderate sensitivity to tuberculin. The remaining 71.2% of the examinees were not infected with tuberculosis; more than 50% of them (41.1%) had postvaccinal allergy. PMID- 15338893 TI - [The new method "cascade" thoracoplasty in surgery of destructive pulmonary tuberculosis]. AB - A new method has been developed for two-stage thoracoplasty with pneumopexy for destructive pulmonary tuberculosis, which is called "cascade" thoracoplasty. The essence of the developed operation is thoracoplasty made in two steps; the objective of Stage 1 is to form limited pleurodesis below the level of lung destruction. At Stage 2, traditional upper posterior thoracoplasty is performed by the method developed by L. K. Bogush. The subjects of the study were 36 patients with destructive forms of pulmonary tuberculosis, including 11 (30.6%), 12 (33.3%), and 13 (36.1%) patients with fibrocavernous, cavernous, and disseminated tuberculosis, respectively. The efficiency of the new developed surgical technique was 83.3% whereas that of traditional thoracoplasty was 78.1% (p < 0.05). According to the clinical forms of pulmonary tuberculosis, the new developed thoracoplastic procedure was effective in 9 (81.8%), 11 (84.6%), and 10 (83.3%) patients with fibrocavernous, disseminated, and cavernous tuberculosis, respectively (p > 0.05). Collapse surgical intervention into the adhesion-free pleural cavity is a basically new technique. PMID- 15338894 TI - [Efficiency of an intensive stage of chemotherapy for tuberculosis in elderly and senile patients]. AB - A hundred new cases of active pulmonary tuberculosis whose age was 60 to 87 years were treated by the intensive chemotherapy regimen recommended by the WHO. Most (n = 78) patients satisfactorily tolerated the intensive stage, fully completed the regimen, and made up a study group (Group 1). Group 2 comprised the remaining 22 patients who had, due to its poor tolerance, to transfer to their individual regimen. The higher incidence of significant cardiovascular diseases in Group 2 patients is the only significant difference between these two identified groups. The advantages of the standard chemotherapy regimen are the more rapid elimination of symptoms of tuberculous intoxication; the earlier and more frequent disappearance of Mycobacterium tuberculosis from the sputum even with the primary drug resistance of the causative agent; as well as a more frequent closure of decay cavities. The findings make it possible to recommend a wider use of the standard intensive chemotherapy regimens for elderly and senile patients with tuberculosis. PMID- 15338895 TI - [Bactericidal therapy in patients with tuberculosis]. PMID- 15338896 TI - [Long-term results of treatment for drug-resistant pulmonary tuberculosis in socially adapted patients]. AB - The 5-year time course of changes in a tuberculous process was studied in 111 patients with drug-resistant pulmonary tuberculosis. 42% of the patients died from tuberculosis. The activity of tuberculosis was preserved in 23% of the patients. Cure occurred in 30%. Cure was achieved in 72% of the patients in case of Mycobacterium tuberculosis monoresistance and in 13.5% in case of multiple resistance to, on the average, 5 drugs, including isoniazid and rifampicin. PMID- 15338897 TI - [Plasmapheresis in the multomodality treatment of active respiratory sarcoidosis]. AB - Eighty-four patients with different stages of respiratory sarcoidosis (RS) were examined. Plasmapheresis was performed during pathogenetic and symptomatic therapy. A comparative analysis was made in the patients from a control group who had no plasmapheresis sessions. The findings have indicated that it is expedient to use plasmapheresis at all stages of active respiratory sarcoidosis. The best results were obtained in young patients with sarcoidosis of intrathoracic lymph nodes and the lung (Stage II). Plasmapheresis promoted a decreased process activity as improved health status in the patients, normalized laboratory parameters, a rapid involution of changes in the bronchopulmonary apparatus, which allowed the doses of corticosteroids to be substantially reduced in the basic therapy. PMID- 15338898 TI - [Experimental evidence for Mycobacterium tuberculosis persistence in M. tuberculosis-infected H37RV mice in the treatment with 3 first-line drugs (rifampicin, isoniazid, pyrazinamide)]. AB - To detect Mycobacterium tuberculosis (MBT) DNA in the blood and organs of tuberculosis-infected animals treated by the standard regimen including 3 first line drugs, to assess the cellular reactions of organs of MBT-infected mice, and to evaluate the effect of rifampicin supplemented to the Cornell treatment regimen on the occurrence of endogenous reactivation of a tuberculous process, 150 BALB/c mice were intravenously infected with the MTB H(3)7Rv strain in a dose of 2.5 x 10(4) CFU/mouse and given chemotherapy with 3 drugs: rifampicin, 12 mg/kg body weight, isoniazid, 5 mg/kg, and pyrazinamide, 35 mg/kg. On day 20 of infection 2, 4, and 6 months after treatment and 2 months after termination of 4- and 6-month courses of therapy, the authors made inoculation, impressionsmears from parenchymatous organs stained by the Romanovsky-Gimse method, and conducted polymerase chain reaction for MBT DNA in the blood and organs of mice. The results indicated that supplementation of rifampicin to the Cornell treatment regimen prevented endogenous reactivation of the process, as confirmed by negative inoculations of the organs of the mice left untreated for 2 months. However, detection of MBT DNA and the cytological picture characteristic of tuberculous inflammation in the blood and organs of mice from these groups suggested that an occult tuberculous process was preserved. PMID- 15338899 TI - [Genotypic characteristics of Mycobacterium tuberculosis strains from the Republic of Tyva]. AB - To determine the genetic heterogenicity of Mycobacterium tuberculosis isolated from patients with tuberculosis from different districts of the Republic of Tyva, 71 M. tuberculosis strains IS6110 according to the polymorphism of restriction fragment lengths were typed; of them 32 strains were spoligotyped. The study could genetically characterize the groups of M. tuberculosis strains circulating among patients with tuberculosis in the Republic of Tyva. There was a predominance of the W family of mycobacteria (60.56%). The spoligotyping permitted identification of M. tuberculosis with the spoligotype Beijing in the Republic of Tyva. The mycobacteria belonging to this spoligotype were prevalent and accounted for 50% of the strains. The high proportion of strains with the unique spoligotype (25%) that have not earlier been encountered in the international database is a specific feature of the Mycobacterium population from the Republic of Tyva. PMID- 15338900 TI - [Radiation damages to the human lung]. PMID- 15338901 TI - Can studies of feral horse behaviour be used for assessing domestic horse welfare? PMID- 15338902 TI - Abdominal adhesions--have we made any progress? PMID- 15338903 TI - Recombinant human erythropoietin and the anaemic horse: flogging a dead horse? PMID- 15338904 TI - Abdominal surgery in foals: a review of 119 cases (1977-1994). AB - The case records of 119 young horses (all less than age one year) that underwent an exploratory celiotomy during a 17 year period were examined to determine the surgical findings, short- and long-term outcome, and prevalence of small intestinal disease compared to previous reports in the mature horse. Physical and laboratory values were compared for long-term survivors vs. nonsurvivors and the frequency of post operative intra-abdominal adhesions was determined. The most common cause for exploratory celiotomy was small intestinal strangulation, followed by enteritis and uroperitoneum. Six horses died during surgery, 23 were subjected to euthanasia at the time of surgery due to a grave prognosis, and 17 horses died or were destroyed after surgery, prior to discharge from the hospital; the short-term survival was 61%. Nine horses were lost to follow-up. Forty-one horses survived long-term (at least 6 months after surgery), 15 died or were subjected to euthanasia after discharge for reasons related to the prior abdominal surgery, and 8 died or were destroyed after discharge due to unrelated reasons, making the long-term survival 45%. Fifty-three (45%) of the horses presented as neonates, and 66 (55%) presented age 3-12 months. Uroperitoneum and meconium impaction were the most common disease in the neonate. Intussusception and enteritis were the most common diseases in older foals. The overall prevalence of small intestinal disease was 44%. Significant elevations in packed cell volume, heart rate, nucleated cell counts and total protein in abdominal fluid and rectal temperature were observed in nonsurvivors compared to survivors. Nonsurvivors had significantly decreased serum bicarbonate, chloride, sodium, and venous pH values. There was no evidence that location of the lesion affected long term survival. Horses with a simple obstruction had a higher survival percentage than those with a strangulating obstruction, and horses that underwent an intestinal resection had a lower long-term survival than those horses undergoing only intestinal manipulation. Nineteen (33%) of the foals examined after the original surgery had evidence of intra-abdominal adhesions. Nine of these (16%) had adhesions that caused a clinical problem. PMID- 15338905 TI - Causes of natal dispersal and emigration and their effects on harem formation in Misaki feral horses. AB - Misaki feral horses were separated into 2 herds and the difference between dispersal from natal group (natal dispersal) and dispersal from natal area (natal emigration) was studied. The causes of dispersal and emigration and their effects on harem formation were studied 1979-1994. The number of horses ranged from 73 (mature males: 8, mature females: 26, young males: 8, young females: 3, colt foals: 6, filly foals: 10 and geldings: 12) in 1979 and 86 (mature males: 14, mature females: 37, young males: 12, young females: 7, colt foals: 5, filly foals: 7 and geldings: 4) in 1994 when the present study ended. All 29 males which survived to age 4 years and 58 females which survived to age 3 years left their natal or mother groups at age one to 3. Seventeen of 22 dispersing males and 29 of 39 dispersing females left their natal groups around the birth of their siblings and significant correlations were found between natal dispersal and birth of a sibling. The number of emigrating young males correlated negatively and significantly with the total number of young males in another herd and the number of emigrating young females correlated positively and significantly with the total number of young females in the natal herd. All 13 emigrating stallions which survived to age 5 years formed stable harem groups and a significant correlation was found between natal emigration and harem formation. Twenty-three of 35 resident mares formed stable consort relations with harem stallions and a significant correlation was found between residence and formation of stable consort relations. PMID- 15338906 TI - Cellular heterogeneity in cathepsin D distribution in equine articular cartilage. AB - The distribution of cathepsin D in normal equine growth cartilage has been examined immunocytochemically using an antiserum raised against human cathepsin D. The cross-reactivity and specificity of the antiserum for equine cathepsin D was confirmed, and its lysosomal localisation was demonstrated in horse skin fibroblasts by confocal scanning microscopy. Cultured horse chondrocytes were heterogenous in their expression of cathepsin D. Heterogeneity of distribution of the enzyme was also seen in chondrocytes in cartilage from different anatomical sites. A high level of cathepsin D was observed in the deep layer of cartilage from the lateral trochlear ridge of the distal femur. Cathepsin D was absent in the hypertrophic zone of the distal radial growth plate. PMID- 15338907 TI - Clinical evaluation of the serodiagnostic value of enzyme-linked immunosorbent assay for Rhodococcus equi infection in foals. AB - An enzyme-linked immunosorbent assay (ELISA) for detection of serum IgG antibodies against Tween 20-extracted antigen of strain ATCC 6939 was applied in Hidaka, Japan to a total of 752 sick foals showing a variety of signs of infectious disease. An optical density (OD) value of more than 0.3 was tentatively fixed to be positive on the basis of readings made of healthy horse sera in previous studies. During a 2 year study, 138 of the 752 sick foals showed an OD value of 0.3 or higher and were designated as 'suspected of R. equi infection'. Age distribution during the initial medical examination of the 138 seropositive foals was significant in that most (64%) foals were age 31-60 days, with a sharp decrease in subjects beyond that age. Of the 138 foals suspected of having R. equi infection, 34 foals (25%) showed OD values of over 0.9 at the initial medical examination, in addition to high blood leucocyte counts and serum fibrinogen and alpha-globulin values. The infectious foals had been treated with antibiotics just before and after serodiagnosis and 126 foals (91%) recovered from the disease. However, no clinical improvement was observed in 12 foals (9%). At necropsy, these foals revealed suppurative pneumonia and lymphadenitis of gut associated lymph nodes accompanied by abdominal abscesses. All isolates from the pulmonary and abdominal abscesses revealed R. equi. These results suggest that OD readings in the high range are associated with severe disseminated infection with R. equi. PMID- 15338908 TI - Clostridium difficile associated with acute colitis in mature horses treated with antibiotics. AB - Clostridium (C.) difficile, or its cytoxin, was demonstrated in faecal samples from 10 of 25 (40%) mature horses investigated with acute colitis treated primarily with antibiotics for disorders other than diarrhoea. C. difficile was not found in faecal samples from 140 horses without signs of enteric disorders, from 21 nondiarrhoeic horses treated with antibiotics, nor from 22 horses with colitis untreated with antibiotics. Except for C. difficile neither Salmonella nor any other investigated intestinal pathogen was isolated in any of the diarrhoeic horses. The findings strongly support some earlier reports that C. difficile is associated with acute colitis in mature horses treated with antibiotics. Of the 10 horses, 4 proved positive for C. difficile both in culture and in the cytotoxin test, 4 in culture only and 2 only in the cytotoxin test. Eight of 10 horses positive for C. difficile were or had recently been hospitalised, indicating that C. difficile may be a nosocomial infection in horses. All horses positive for C. difficile were treated with beta-lactam antibiotics. PMID- 15338909 TI - Prostaglandin F2alpha release associated with an embryo transfer procedure in the mare. AB - The pattern of the main metabolite of prostaglandin (PG) F2alpha was recorded following a nonsurgical embryo transfer technique in 9 mares under field conditions in Estonia. Three patterns were observed. Two of them were characterised by PG release, thereas the third was not. A tendency towards a shortened cycle was seen in 3 mares. Observations were made regarding the manipulation of the uterus as being normal or difficult to perform. In general, mares where the procedure was considered difficult were also found to have a PG release. PMID- 15338910 TI - A survey of reproductive performance in Thoroughbred mares and morbidity, mortality and athletic potential of their foals. AB - A survey was performed to evaluate the reproductive performance of Thoroughbred mares, estimate risks of dystocia and of morbidity and mortality in foals during the first year post partum and their physical acceptability at age one year. The study population consisted of registered Thoroughbred mares and their foals owned by residents of 4 Western Canadian provinces. Owners were identified using information obtained from the North American Jockey Club, and questionnaires were mailed regarding mares bred in 1988 and their foals born in 1989. Eighty-three per cent of mares were reported to be pregnant at some stage following breeding, and 80% of pregnant mares subsequently gave birth to live foals. Estimates of morbidity and mortality were greater than previously reported, 25% of foals had health problems and 5% died during the first 2 weeks postpartum. Twenty-seven per cent of foals surviving 2 weeks were reportedly affected by some health problem between age 15 days and one year, and 6% died during this period. The case fatality rates of horses with upper respiratory tract infections and diarrhoea were much lower than case fatality rates for infectious diseases occurring less frequently. The rate of death or euthanasia among horses with musculoskeletal problems was relatively high after age 2 weeks. Foals with health problems up to age 2 weeks, or between age 15 days and one year were 5 to 7 times more likely to be classified as physically unacceptable for athletic use. Angular limb deformity was the health problem most commonly reported in foals receiving unacceptable physical assessments, and assessments of longterm athletic potential were apparently not affected by the occurrence of infectious diseases. PMID- 15338911 TI - Pulmonary vascular pressures of strenuously exercising Thoroughbreds after administration of varying doses of frusemide. AB - The frusemide dose-response for attenuation of exercise-induced pulmonary capillary hypertension was studied in 7 healthy, exercise-conditioned Thoroughbred horses using previously described haemodynamic procedures. Four different doses of frusemide were tested: 250 mg regardless of bodyweight (amounting to 0.56 +/- 0.03 mg/kg bwt), 1.0 mg/kg bwt, 1.5 mg/kg bwt and 2.0 mg/kg bwt. Frusemide was administered i.v., 4 h before exercise. Haemodynamic data were obtained at rest and during treadmill exercise performed at 14.2 m/s on a 3.5% uphill grade; this workload elicited maximal heart rate of horses. Airway endoscopy was performed post exercise to detect exercise-induced pulmonary haemorrhage (EIPH). In standing horses, frusemide administration resulted in a significant (P<0.05) decrease in mean pulmonary arterial, pulmonary capillary and pulmonary artery wedge pressures, but significant differences among the various frusemide doses were not observed. In the control experiments, exercise caused significant increments in the right atrial as well as pulmonary arterial, wedge, and capillary pressures, and all horses experienced EIPH. Following frusemide administration, the exercise-induced rise in right atrial and pulmonary vascular pressures was significantly attenuated, but significant differences between the frusemide doses of 250 mg, 1.0 mg/kg, and 1.5 mg/kg were not discerned and all horses remained positive for EIPH. Although a further significant (P<0.05) attenuation of the exercise-induced rise in pulmonary capillary blood pressure occurred when frusemide dose increased from 250 mg to 2.0 mg/kg bwt, all horses still experienced EIPH. It is concluded that a linear response to increasing frusemide dosage in terms of attenuation of the pulmonary capillary hypertension does not exist in strenuously exercising Thoroughbred horses. PMID- 15338912 TI - Tachykinin receptors in the equine pelvic flexure. AB - Tachykinins, of which substance P (SP) is the prototype, are neuropeptides which are widely distributed in the nervous systems. In the equine gut, SP is present in enteric nerves and is a powerful constrictor of enteric muscle; in other species, SP is also known to have potent vasodilatory and pro-inflammatory effects. The specific effects of SP are determined by the subtype of receptor present in the target tissue. There are 3 known subtypes of tachykinin receptors, distinguished by their relative affinities for SP and other tachykinins. The distribution of SP binding sites in the equine pelvic flexure was determined using 125I-Bolton Hunter SP (I-BHSP) autoradiography. Most I-BHSP binding sites were determined to be saturable and specific, therefore presumably representing tachykinin receptors. The greatest degree of I-BHSP binding occurred over very small vessels, and over the muscularis mucosae; I-BHSP binding was also intense over the circular muscle of the muscularis externa and mucosa, and present, although less intense, over the longitudinal muscle of the muscularis externa. Competition of I-BHSP with specific receptor agonists for binding sites in the equine pelvic flexure were used to determine the subtypes of tachykinin receptors present. The neurokinin-1 receptor subtype predominated in the equine pelvic flexure, followed by the neurokinin-3 receptor subtype. PMID- 15338913 TI - The association of erythromycin ethylsuccinate with acute colitis in horses in Sweden. AB - In Sweden there are several reports of mares developing acute colitis while their foals were being treated orally for Rhodococcus equi pneumonia with the combination of erythromycin and rifampicin. In this study 6 adult horses were given low oral dosages of these antibiotics, singly or in combination. Within 3 days post administration of erythromycin, in one case in combination with rifampicin, 2 horses developed severe colitis (one fatal). Clostridium difficile was isolated from one of the horses, whereas no specific pathogens were isolated from the other. Both horses had typical changes in blood parameters seen in acute colitis. Clostridium difficile was also isolated from the faeces of a third horse given an even lower dosage of erythromycin in combination with rifampicin. This horse developed very mild clinical symptoms and recovered spontaneously. In the fourth horse given erythromycin only, very high numbers of Clostridium perfringens were isolated. The horses given rifampicin only did not develop any clinical symptoms and there were no major changes in their faecal flora. In conclusion, it has been demonstrated that low dosages of erythromycin ethylsuccinate can induce severe colitis in horses associated with major changes of the intestinal microflora. Clostridium difficile has been demonstrated as a potential aetiological agent in antibiotic-induced acute colitis. PMID- 15338914 TI - Epidemiology of aminoglycoside resistance in a large animal hospital. PMID- 15338915 TI - Eosinophilic enterocolitis associated with recurrent colonic impactions in a mare. PMID- 15338916 TI - Nonregenerative anaemia associated with administration of recombinant human erythropoietin to a Thoroughbred racehorse. PMID- 15338917 TI - Address of Mrs. Satish Chawla, President, TNAI delivered at the inaugural function of XX TNAI Biennial (69th) Conference, Jaipur, on November 24, 2003. PMID- 15338918 TI - A study of correlates of Acute Respiratory tract Infection (ARI) among infants in a selected area of Udupi District. PMID- 15338919 TI - Nursing care of patient with obstructive sleep apnoea. PMID- 15338920 TI - Nurse's responsibilities in phototherapy. PMID- 15338921 TI - Fast track to nursing. One university is about to introduce an innovative course offering registration to graduates within two years. PMID- 15338922 TI - Writer's block. Nurses are exasperated by the slow pace at which prescribing is expanding and the limitations on what they can prescribe. PMID- 15338923 TI - A release from suffering. Supporters of a bill being introduced to parliament hope to make assisted dying legal while still protecting the vulnerable. PMID- 15338924 TI - Safe as houses. Living on a hospital site may be convenient but there are potential security risks. PMID- 15338925 TI - When I'm sixty five. A public sector pension has always been an attractive benefit for nurses. But, as Sue Ward reports, a review of the NHS scheme could change all that. PMID- 15338926 TI - Nature's cure. PMID- 15338927 TI - Breathing difficulties. Vikki Robinson learned more about assessing breathless patients after reading a continuing professional development article. PMID- 15338928 TI - Listen carefully. We listen, but all too often we do not really hear. PMID- 15338929 TI - Narrative art inquiry and anorexia nervosa. AB - Narrative art inquiry is a new qualitative research methodology. It is different from the narrative approaches used by some nurse investigators because the study's findings are presented in the form of creative writing. In this article the author describes narrative art inquiry using material from research that used this approach to study anorexia nervosa. The author believes that narrative art inquiry has a wider application and could be used to study other types of psychological illness and physiological distress. PMID- 15338930 TI - Sexual health: meeting adolescents' needs. AB - BACKGROUND: The UK has the highest rate of sexually transmitted infections (STIs), especially among young people, in western Europe. Knowledge of what influences young people's sexual behaviour is limited which highlights a need to understand adolescent development and recognise that they are at increased risk of acquiring an STI compared with other groups. This article examines the risks of acquiring STIs in adolescence and discusses how nurses feel about treating young people with sexual health needs. CONCLUSION: Postgraduate training needs to incorporate specific modules relating to young people within sexual health. Sexual health providers go some way to providing an adolescent-friendly service, but more creative approaches to service delivery such as flexible opening hours and contraception outreach services would make them more accessible to young people. PMID- 15338931 TI - Diabetic emergencies in adults. AB - In this article the author discusses three diabetic emergencies: diabetic ketoacidosis, hyperosmolar non-ketotic syndrome and hypoglycaemia, which all require prompt recognition and appropriate intervention. PMID- 15338932 TI - Piecing together the perfect leader for England's nurses. Nurses expect a lot of the next chief nursing officer for England. PMID- 15338933 TI - Reducing junior doctors' hours will extend opportunities for nurses. PMID- 15338934 TI - New ethical recruiting code will not end exploitation. PMID- 15338935 TI - Every nurse should have leadership skills. PMID- 15338936 TI - Reduced doctors' hours. PMID- 15338937 TI - Home is where the nurse is. PMID- 15338938 TI - Healing the scars. PMID- 15338939 TI - Understanding the long-term challenges of cancer survivorship. AB - Survivorship in the context of cancer is an expanding domain. It is important not just to eradicate the cancer but also to understand the implications of diagnosis and treatment. This may also include a fear of secondary malignancies and life threatening complications. PMID- 15338940 TI - What you need to know about... Tinea pedis. PMID- 15338941 TI - Achieving best practice in routine observation of hospital patients. AB - This article reviews the evidence supporting best practice when undertaking patient observations in a district general hospital. Correct monitoring of the patients' progress is essential in the detection of adverse events and can potentially prevent admission to intensive care or high dependency units. PMID- 15338942 TI - Benchmarking patient privacy to improve essence of care. AB - Concerned about the privacy offered to patients at a large teaching hospital in the west Midlands, staff established a multidisciplinary group to benchmark privacy and dignity. They have ensured a high standard of care for patients through the benchmarking process. This article details the steps taken by the group and illustrates how Essence of core has successfully provided a framework to support quality improvements. PMID- 15338943 TI - Causes and treatment of neonatal respiratory distress syndrome. AB - Neonatal respiratory distress syndrome is a condition of advancing respiratory distress, commencing at or shortly after birth. The disease follows an acute course, with deterioration within 48 hours followed by stabilisation and improvement. Treatment is to support respiratory function and maintain good oxygenation. PMID- 15338944 TI - Understanding the impact of malignant mesothelioma. PMID- 15338945 TI - How to set up a learning network. The process of creating a nurse forum or network can be just as valuable to your continuing professional development as the meetings that follow. PMID- 15338946 TI - Is being a customer something to aspire to? PMID- 15338947 TI - Editorial overview: Transporter proteins in drug disposition and uptake. PMID- 15338948 TI - Virtual screening in lead discovery and optimization. AB - Virtual screening by molecular docking, using a protein with an experimentally determined structure as a target, has become an established method for lead discovery and for enhancing efficiency in lead optimization. Generalizations of the quantitative structure-activity relationship concept have led to approaches for virtual screening in the absence of a protein target structure, instead relying upon ligand-based models as surrogates of protein active sites. Recently reported methods for ligand-based virtual screening can achieve similar enrichment rates to those obtained using molecular docking. This review will discuss recent advances in both domains of virtual screening, including theoretical and practical advances and the implications for their application. PMID- 15338949 TI - Structure-guided fragment screening for lead discovery. AB - Fragment-based ligand screening can be a highly effective strategy for drug discovery. In general, fragment hits interact efficiently with the target, and although the potency of these small binders is often low, their optimization into potent leads is tractable. For a hit optimization phase to take full advantage of a good quality fragment binder, we believe it is essential to obtain reliable structural data for the hits. In this review, we describe the methods used for structure-based fragment screening and fragment-to-lead optimization and discuss a number of applications from the literature. PMID- 15338950 TI - Affinity-based screening techniques for enhancing lead discovery. AB - Contemporary, rational small-molecule lead discovery methods, comprising target identification, assay development, high-throughput screening (HTS), hit characterization and medicinal chemistry optimization, dominate early-stage drug discovery strategies in many pharmaceutical companies. There is a growing disparity between the increasing cost of funding these methods and the decreasing number of new drugs reaching the market. New strategies must be adopted to reverse this trend. The use of genomics- and proteomics-based target discovery efforts can aid the process by dramatically increasing the number of novel, more highly validated targets entering the discovery process, but HTS must meet this increased demand with faster, cheaper technologies. Although activity-based screening strategies are typically efficient, allowing one scientist to interrogate tens of thousands of compounds per day, affinity-based screening strategies can allow much greater efficiency in the overall process. Affinity based methods can play a role in both facilitating the screening of a greater number of targets and in efficiently characterizing the primary hits discovered. PMID- 15338951 TI - Microwave-enhanced medicinal chemistry: a high-speed opportunity for convenient preparation of protease inhibitors. AB - The unique properties of microwave in situ heating offer unparalleled opportunities for medicinal chemists to accelerate lead optimization processes in early drug discovery. The technology is ideal for palladium-catalyzed alteration chemistry as it allows for complete control over reactions, with the use of non inert conditions, providing high chemoselectivity and rapid feedback. To illustrate the advantages of this methodology, we describe our applications and approaches for the rapid synthesis of novel aspartyl protease inhibitors using dedicated microwave equipment. Biological results from chemical studies of the different side-chain positions of HIV-1 and malarial plasmepsin I and II protease inhibitors are summarized. PMID- 15338952 TI - The role of structure in kinase-targeted inhibitor design. AB - Protein kinases are attractive targets for rational drug design against a wide range of diseases. From detailed knowledge of the structure-function relationships underlying protein kinase activity and regulation, a number of methods for achieving kinase inhibition have been suggested and explored using structure-aided drug discovery. Attaining selective protein kinase inhibition in a cellular context, and converting the large number of known potent kinase inhibitors into effective drugs, are outstanding problems in this area and, from a structural perspective, the challenges presented by modulating pharmacokinetics and minimizing the incidence of resistant mutations in the target are of particular interest. PMID- 15338953 TI - Transforming the TGFbeta pathway: convergence of distinct lead generation strategies on a novel kinase pharmacophore for TbetaRI (ALK5). AB - The pathological activation of the transforming growth factor beta (TGFbeta) pathway plays a critical role in the progression of fibrotic diseases and also enhances tumor invasiveness and metastasis. Due to its central role in TGFbeta signaling, the TGFbeta type I receptor (TbetaRI) is emerging as an exciting target for blockade of the TGFbeta pathway. In this review we will discuss how three independent drug discovery strategies, ie, target-hopping, high-throughput screening and virtual screening, have converged in the identification of inhibitors of TalphaRI kinase. Structural studies have provided insight into the potency and selectivity of these inhibitors and form the basis for structure based design optimization strategies. These efforts have enabled the production of potent, selective inhibitors for dissecting the TGFalpha pathway and assessing the usefulness of TalphaRI blockade in the treatment of fibrotic diseases and cancer. PMID- 15338954 TI - Progress and development of small molecule HCV antivirals. AB - Hepatitis C virus (HCV) is a disease that has a growing impact worldwide. A combination therapy comprising interferon-alpha (IFNalpha) and ribavirin represents the current standard treatment for chronic HCV infection, although it has demonstrated limited success and causes some serious side effects. Promising alternative approaches toward the control of HCV infection, and the development of new antiviral agents, include the use of NS3/4A serine protease and NS5B polymerase inhibitors. Successful proof-of-concept clinical trials of the NS3/4A protease inhibitor BILN-2061 have confirmed the usefulness of a peptidomimetic product-based approach, providing impetus for the generation of improved molecules. Preclinical results from the development of HCV polymerase inhibitors, both nucleoside and non-nucleoside, are promising. This review provides an overview of recent progress in these areas, and discusses the potential of various approaches toward small molecule HCV antivirals. PMID- 15338955 TI - The race to an orally active Factor Xa inhibitor: recent advances. AB - Factor Xa (fXa) is a key enzyme in the coagulation cascade and an essential component of the prothrombinase complex, which activates prothrombin to thrombin, leading to fibrin clot formation. In the search for a more effective and safer orally active anticoagulant, fXa has emerged as a major target for potential therapeutic applications in the treatment and prevention of thrombosis. This review focuses on recent advances in the chemistry of drug design and lead optimization of orally bioavailable fXa inhibitors. Many of these orally active fXa inhibitors are currently in clinical trials and are anticipated to change the landscape of thrombosis therapy. PMID- 15338956 TI - Molecular properties that influence oral drug-like behavior. AB - Pharmaceutical companies are constantly racing to discover the next therapeutic blockbuster. The consensus in the industry is to focus on compounds that are by some measure drug-like, but in order to do this effectively a number of questions must be answered. For example, how should drug-like be defined and how might this definition be used to enhance drug discovery? Has the field moved beyond Lipinski's seminal 'rule-of-five' observations? This review offers a working definition of oral drug-likeness, describes various approaches used in its characterization and discusses its appropriate use. We will focus primarily on the use of computed molecular properties that attempt to predict the oral drug like behavior of compounds and propose guidelines for the use of observations and trends established in existing datasets to support drug discovery efforts. In particular, this review will demonstrate how trends in simple properties of the data can be used prospectively to compare and prioritize groups of compounds, chemical libraries and different chemical series with greater reliability than for predicting drug-likeness of single compounds. It is the authors' belief, however, that properties or descriptors that will completely separate drug-like space from non-drug-like space are unlikely to be found; the focus should instead lie on overall distributions of drug-like and non-drug-like compounds in the property space that tend to overlap significantly. PMID- 15338957 TI - Farnesyltransferase inhibitors as anticancer agents: critical crossroads. AB - Farnesyltransferase (FT) inhibitors were originally designed as anticancer agents, and were thought to act by inhibiting the farnesylation of mutant Ras proteins. However, these compounds were subsequently demonstrated to have antitumor effects even in the absence of Ras mutations and it has now become clear that other protein targets are involved. This article discusses the preclinical and clinical development of FT inhibitors. To date, tipifarnib (Zarnestra; Janssen Pharmaceutica NV) and lonafarnib (Sarasar; Schering-Plough Research Institute) are the only two FT inhibitors to have been evaluated in phase III clinical trials. The clinical results of these two compounds are presented below, with emphasis on ways of enhancing the possibility of a successful FT inhibitor anticancer drug. Details of new FT inhibitors disclosed since the beginning of 2003 are also included. PMID- 15338958 TI - Corticotropin-releasing factor antagonists: recent advances and exciting prospects for the treatment of human diseases. AB - Corticotropin-releasing factor (CRF) co-ordinates the neural, endocrine and immune responses of the body to stress. Several studies have implicated CRF in the etiology of anxiety, depression, substance abuse, stress-related gastrointestinal disorders and preterm labor, and intensive research into the design of safe and effective CRF antagonists is currently being pursued in several laboratories. Recently, improvements have been made not only in brain penetrance and in vivo activity in preclinical models for anxiety, depression and irritable bowel syndrome, but also in structural diversity for these compounds. Clinical data for R-121919 (NBI-30775; Neurocrine Biosciences Inc) raises the expectation that safe and potent CRF antagonists might be useful as drugs for the treatment of human diseases. PMID- 15338959 TI - Recent advances in CB1 cannabinoid receptor antagonists. AB - Cannabinoid CB1 receptor antagonists are currently the subject of intensive research due to their highly promising therapeutic prospects. Novel chemical entities having CB1 antagonistic properties have recently been disclosed by several pharmaceutical companies and some academic research groups, some of which are close structural analogs of the leading compound rimonabant (SR-141716A; Sanofi-Synthelabo). A considerable number of these CB1 antagonists are bioisosteres that are derived from rimonabant by the replacement of the pyrazole moiety with an alternative heterocycle. As well as these achiral compounds, Solvay Pharmaceuticals have disclosed a novel class of chiral pyrazolines that are potent and CB1/CB2 subtype-selective cannabinoid receptor antagonists, in which the interactions with the CB1 receptor are highly stereoselective. PMID- 15338960 TI - Predicting plasma protein binding of drugs--revisited. AB - Plasma protein binding of drugs has been studied for almost 100 years, but despite the accumulation of large amounts of data, the accurate prediction of this ADME parameter continues to be problematic. This review outlines recent efforts on the development of prediction tools for plasma protein binding of drugs, specifically human serum albumin, in the context of its relevance and its influencing factors. The issue of why it is difficult to achieve prediction of sufficient quality for a diverse dataset will also be considered. PMID- 15338961 TI - Recent advances in the design of matrix metalloprotease inhibitors. AB - Inhibition of matrix metalloproteases (MMPs) for the treatment of diseases, such as cancer, arthritis and other diseases associated with tissue remodeling, has become an area of intense interest in the pharmaceutical industry in recent years. Despite tremendous efforts over the last decade to explore individual members of this target family, along with multiple inhibitor classes, simple and effective drugs for inhibiting individual MMPs have not yet emerged. This review highlights the major developments in research into MMPs and their inhibitors, from the recent medicinal chemistry literature, with a focus on structure-based design, selectivity and pharmacokinetic (PK) properties. The increasing availability of high-resolution X-ray crystal structures for many members of this protein family makes MMPs ideally suited for structure-based design approaches, which are now routinely used in this area. The most challenging aspect of lead optimization for MMP inhibitors is in finding candidates having acceptable pharmacological, PK and selectivity profiles. Clinical trials in cancer giving disappointing results have led to discussions on how to gain adequate MMP selectivity in order to minimize side effects. Unfortunately, careful analysis of X-ray crystal structures has not suggested any simple solutions. These areas collectively constitute the main challenges in the current search for orally available MMP inhibitors, and will be discussed in this review. PMID- 15338962 TI - Design and development of BACE-1 inhibitors. AB - In early 1999, beta-amyloid cleaving enzyme-1 (BACE-1) was identified as the protease responsible for the critical first step in the processing of beta amyloid precursor protein that ultimately leads to the production of Abeta peptides in the brain. Accumulation of these peptides has been implicated in the pathology of Alzheimer's disease (AD). An inhibitor of BACE-1 would therefore have therapeutic potential to slow or halt the progression of this debilitating and ultimately fatal disease. This review provides a perspective on the recent developments in the design of BACE-1 inhibitors. An overview of early research is also included, with particular emphasis on a comprehensive survey of the patent literature. PMID- 15338963 TI - Factors responsible for needlestick injuries to nurses. PMID- 15338964 TI - Infection control nurses as the professional voice of infection control nursing. PMID- 15338965 TI - A study to assess the knowledge and attitude of unmarried young adults about desirable sex behaviour. PMID- 15338966 TI - How to choose the right banker... AB - Avenues for obtaining capital are numerous, affording you more choices for financing than ever before. Loaned dollars have become a commodity in the opinion of many, so the options for borrowers can be overwhelming. In all cases, I would submit to you, the right banker makes a difference. With that being said, what traits should you look for in selecting the right banker? The characteristics below will help guide you down that path. PMID- 15338967 TI - Best management practices for amalgam waste. PMID- 15338968 TI - Oral pathology. Lichen planus. PMID- 15338969 TI - [Treatment of multiple myeloma with intravenous pamidronate. Pain prevention and suppression of hypercalcemia risk]. AB - In a prospective clinical study, with the patient as its own control, we selected patients with stage III multiple myeloma. We treated them monthly with intravenous (i.v.) Disodic Pamidronate: 90 mg in 2 to 21 cycles. Four patients died during the study. The remaining 13 patients presented reduced bone resorption urinary markers (D-Pyr mainly) as well as urinary calcium (bone turnover reduction). Both effects (metabolic interchange reduction and calcium variation) did not show a direct relationship, being the 1st magnitude proportional to the baseline level and the 2nd independent from it. We noted pain reduction (VAS: visual analogs scale), low analgesic consumption, and the absence of future skeletal problems. The treatment tolerance was good. All these factors contribute to justify the welfare of the patient demonstrated by ECOG (Eastern Cooperative Oncology Group), not only improving the average results but also extending this improvement to future results. Our observation suggests that under strict procedures, this treatment could be very adequate in patients with advanced multiple myeloma independent of the state of the disease at the beginning of the study. PMID- 15338970 TI - [Comparison between electrocardiographic test with dobutamine and exercise stress testing on coronary risk stratification]. AB - The exercise testing is still the most common test used to stratify the patients's risk of new events following an acute myocardial infarction, but about 20 to 40% of the patients can not perform it appropriately. Since the electrocardiographic test with dobutamine has proved to be easy and safe, our aim was to evaluate its capacity to predict the results of the exercise testing on patients after an acute myocardial infarction. A total of 210 patients (average of age 60.5+/-11.7 years old and 23.3% females) recovering from a first uncomplicated myocardial infarction, were consecutively included. An electrocardiographic test with dobutamine was performed during the fifth day of admission, and an exercise test during the sixth. The development of chest pain and/or a descending ST segment > or = 1 mm during a test qualified it as positive for ischemia. The sensitivity, specificity, positive and negative predictive values of both tests were determinated, as well as the congruence on their results, a p-value < 0.05 was considered as statistically significative. Whereas 91 patients achieved a positive result on the electrocardiographic dobutamine tests, 54 patients were positive during exercise. On the other hand, both tests came up positive in 49 patients, and negative in 114 patients. The hemodynamic variables were similar during both tests, except in those patients with negative results with both methods, who developed a higher maximal arterial pressure and double product during exercise. Accordingly, the electrocardiographic dobutamine test showed a sensivity 90.17%, a specificity 73%, positive and negative predictive values 54% and 95.8%, respectively, to predict the results of the exercise testing, with a Kappa value of 0.52. In conclusion, the electrocardiographic dobutamine test showed not only to predict the results of the exercise testing adequately with a high negative predictive value, but also to be safe, a useful and simple diagnostic tool for the evaluation of patients after an acute myocardial infarction. PMID- 15338971 TI - [High doses of aspirin reduce natriuresis in hypertensive patients treated with enalapril]. AB - Angiotensin converting enzyme inhibitors have been shown to be useful in the treatment of essential hypertension while anti-platelet agents improve the overall cardiovascular risk profile in this population. Our aim was to assess the interaction of two different aspirin (ASA) doses--81 and 325 mg/day--with the antihypertensive effect of enalapril as well as their impact upon the urinary sodium excretion (Na(u)). A total of 22 patients between 35 and 65 years of age were included in a prospective double blind trial with a partial cross-over design. We excluded patients with secondary hypertension and recent use of anti inflammatory drugs. Patients were placed on enalapril and a low sodium diet--<6 g of NaCl/day--and, sequentially, on two different doses of aspirin separated by a 10 day wash out period. Blood pressure (BP) was measured at weekly visits. Systolic, diastolic and mean BP levels decreased significantly in enalapril treated patients (p<0.01) and no difference was detected between the two AAS dosages although a non-statistically significant difference towards better BP control was observed when 81 mg of ASA was used. Na(u) was higher at baseline when compared with the two periods under ASA (p<0.01) and Na(u) was higher with 81 mg than with 325 mg. These results suggest that in essential hypertensive individuals treated with enalapril and two ASA doses, low doses of ASA are associated with better blood pressure control and higher natriuresis. PMID- 15338972 TI - Evaluation of phenotypic and genotypic markers for clinical strains of Acinetobacter baumannii. AB - Acinetobacter baumannii is an important opportunistic pathogen that is rapidly evolving toward multidrug resistance and is involved in various nosocomial infections that are often severe. It strongly prompts the epidemiological study of A. baumannii infections. However, there is no a generally accepted typing scheme. Different genotypic and phenotypic procedures were evaluated for the characterization of clinical isolates of A. baumannii isolated from a community hospital of Rosario, Argentina (Hospital de Emergencias Clemente Alvarez, HECA), during a period of four years. These included PCR with degenerate oligonucleotide primers (DO-PCR), repetitive extragenic palindromic-PCR (REP-PCR), pulsed-field gel electrophoresis (PFGE), and antibiotyping. Amongst individual methods, DO-PCR and PFGE were found the most suitable methods to discriminate A. baumannii clinical isolates [discriminatory indexes (D) of 0.98 and 0.96, respectively]. On the other hand, both antibiotyping and REP-PCR were much less discriminatory (D: 0.86 and 0.77, respectively). The combination of antibiotyping with any of the above genotypic procedures was found to largely increase D. In particular, the combination of DO-PCR and antibiotyping provided the best discriminatory method for epidemiological studies of A. baumannii. Combination of the different genotypic and phenotypic procedures allowed the inference of genetic relationships and dissemination of multidrug-resistant A. baumannii clones in HECA in the period 1994-1999. One particular strain, which showed sensibility to carbapenems, was found widely distributed in this hospital during 1994-1996. A different strain, showing additional resistance to carbapenems, rapidly disseminated in HECA in coincidence with the introduction of imipenem therapy in 1997. PMID- 15338973 TI - [Usefulness of erythrocyte protoporphyrin test in the puerperium compared to the soluble transferrin receptor]. AB - Erythrocyte protoporphyrin (EP), serum ferritin (SF), soluble transferrin receptor (sTfR) and routine hematological laboratory tests were studied in 77 women 24 h post-partum, assisted at Paroissien Hospital (in Buenos Aires Province). Hematocrite (Hct), hemoglobin (Hb), red blood cells (RBC) and white blood cells (WBC) were determined using an electronic counter (Mega); EP by Piomelli's; SF by ELISA (IMx Ferritina, Abbott); sTfR by ELISA (Orion Diagnostica) and C-Reactive Protein (PCR-Latex, Wiener lab). All determinations were made in fasting blood samples. Statistical analysis (Receiver Operating Characteristics, ROC) were performed using Med Calc software. The soluble transferrin receptor (sTfR) was used as gold standard. RESULTS: (mean +/- SD): Hct (%) 35 +/- 5; Hb (g/l) 113 +/- 18; RBC x 103/mm3 3,893 +/- 489; MCV (fL) 90 +/- 6; WBC/mm3 9,543 +/- 2,669; EP (microg/dl RBC) 46 +/- 39; sTfR (mg/l) 4.7 +/- 2.8; SF (microg/l) 26 +/- 31; PCR (Pos/Neg) 72/5. EP did not correlate with SF but it did with sTfR (r=0.323, p=0.007). Sensitivity (Se) and specificity (Sp) of SF and EP were, respectively: 83% and 63% for a cut-off point of 25 microg SF/L; 38% and 90% for a cut-off point of 53 microg EPdl RBC. These results suggest that EP might be a useful and cheap indicator to assess maternal Fe nutritional status during the early perinatal period in hospital laboratories, allowing the detection of 16% of women presenting normal Hb values. PMID- 15338974 TI - [Renal injury in rats poisoned by venom of Bothrops neuwiedii diporus from Argentina]. AB - Acute renal failure is one of the systemic complications that can be found in bothropic accidents. In this study the effects on male Wistar rats induced by the venom of Bothrops neuwiedii diporus were evaluated. The histopathology revealed acute tubular necrosis, lesions firstly were observed 3 hours post inoculation of 700 microg of venom. Cortical kidney congestion and granulohialin degeneration of tubular epithelial cells were observed, these lesions achieved a maximum at 24 hours after inoculation. Tubular epithelial hidropic degeneration and dilatation of tubular lumen with hyalin casts were present inclusive up to 4 weeks after inoculation. Biochemical parameter values associated with kidney renal failure were increased 6 hours after venom inoculation (urea: 1.10+/-0.22 g/dl; creatinine: 19.60+/-1.51 mg/dl), but at the end of the first week they decreased till normal values. The urinary density was lower than normal value: 1.005+/ 0.001 (p<0.001) and at the end of the first month they oscillated between 1.005 and 1.060 (p<0.001). Renal injury induced by B. neuwiedii diporus could be better appreciated by histopathology than by the routine laboratory assays. PMID- 15338975 TI - [Osteonecrosis in HIV-infected patients]. AB - Osteonecrosis, also known as avascular necrosis, is chiefly characterized by death of bone caused by vascular compromise. The true incidence of osteonecrosis in HIV-infected patients is not well known and the pathogenesis remains undefined. Hypothetical risk factors peculiar to HIV-infected individuals that might play a role in the pathogenesis of osteonecrosis include the introduction of protease inhibitors and resulting hyperlipidemia, the presence of anticardiolipin antibodies in serum leading to a hypercoagulable state, immune recovery and vasculitis. Hereby we present a series of 13 HIV-infected patients with osteonecrosis. The most common symptom upon presentation was arthralgia. The majority of the patients had received steroids, 9 had developed hyperlipidemia after the introduction of HAART, 8 were smokers and 4 patients were alcoholics. In 2 patients, seric anticardiolipin antibodies were detected. Twelve patients had AIDS and were on HAART (11 were on protease inhibitors). We believe that osteonecrosis should be included as differential diagnosis of every HIV-infected patient who complains of pain of weight bearing joints. Likewise, it seems prudent to rule out HIV infection in subjects with osteonecrosis. PMID- 15338976 TI - Oleoyl-estrone metabolic effects in relation with caloric restriction in inbred Beta rats with spontaneous obesity and type 2 diabetes. AB - Spontaneously hypertriacylglycerolemic obese and diabetic inbred IIM Beta rats were treated with oleoylestrone for 10 days. Pair-feeding was performed to determine some oleoyl-estrone effects dependent on the caloric restriction it promotes. Twenty-five 200 day-old Beta males receiving a daily gavage of 0.2 ml sunflower oil were divided into the following groups: 1) daily dose of 10 nmol/g oleoyl-estrone; 2) pair-fed; 3) control. The variables measured were: whole body protein, water and lipid; retroperitoneal and epididymal fat depot weights; plasma urea, glucose, insulin, triacylglycerols and cholesterol. Biomass and food intake were assessed daily. Oleoyl-estrone and pair-fed groups expressed similar variations in body composition and significant body weight losses due to reduction in food intake. Oleoyl-estrone and pair-fed treatments significantly reduced retroperitoneal fat depot weights, but not epididymal ones. In oleoyl estrone and pair-fed groups hyperglycemia decreased and insulinemia lowered significantly. Plasma normal total cholesterolemia and hypertriacylglycerolemia values typical of Beta rats decreased strongly compared to controls, though attaining significantly different values between oleoyl-estrone and pair-fed groups. Plasma total cholesterol appeared as more sensitive to caloric restriction than triacylglycerols through a specific oleoyl-estrone-mediated effect. PMID- 15338977 TI - [Hypocalciuric hypercalcemia due to de novo mutation of the calcium sensing receptor]. AB - The aim of this paper is to refer the unusual case of a 34 years old woman who consulted because of asymptomatic hypercalcemia, detected in a biochemical routine examination. The elevated values of serum calcium without blunted parathyroid hormone secretion suggested a parathyroid pathology. The concomitance of hypocalciuria with hypercalcemia and a calcium clearance/creatinine clearance ratio less than 0.01 reverted the diagnosis of familial hypocalciuric hypercalcemia, the first option. The absence of familial background led to the molecular study of the patient and her family. The latter confirmed the diagnosis of a de novo inactivating mutation of the calcium sensing receptor. Details on the molecular study and a brief review of this subject are included. PMID- 15338978 TI - [Purulent pericarditis with pericardial tamponade caused by Streptococcus agalactiae and Salmonella enterica no typhi]. AB - Purulent pericarditis (PP) is an uncommon condition with high mortality. In the preantibiotic period, Staphyloccocus aureus and Streptococcus pneumoniae were the most common etiologic agents. We describe the case of a 75-year old man with septic shock, PP and cardiac tamponade caused by Streptococcus agalactiae and Salmonella enterica no-typhi. To our knowledge this association of pathogenic organisms has not been previously reported in the literature. The pathogenesis is here reviewed, and in our patient presumably, purulent pericarditis occurred via hematogeneus spread undergoing upper gastrointestinal endoscopy. The patient's course was complicated and he died on 34th hospital day. After this case report it is considered that differential etiologic diagnosis of PP should include these agents, especially in immunodepressed patients with predisposing factors. PMID- 15338979 TI - [Idiopathic retroperitoneal fibrosis]. PMID- 15338980 TI - [Pneumocystis jiroveci (carinii) pneumonia undergoing cavitation]. PMID- 15338981 TI - [Systemic vasculitis with lung damage and severe sepsis]. PMID- 15338982 TI - [Enterohemorrhagic Escherichia coli and hemolytic-uremic syndrome in Argentina]. AB - The hemolytic-uremic syndrome (HUS) is a multisystemic disorder that is characterized by the onset of acute renal failure, microangiopathic hemolytic anemia and thrombocytopenia. It is the most common cause of acute renal failure and the second cause of chronic renal failure and renal transplantation in children in Argentina. Our country has the highest incidence of HUS in the world, with approximately 420 new cases observed each year with an incidence of 12.2 cases per 100,000 children in the age group 0-5 years. Numerous etiologic factors have been associated with HUS but the infection with enterohemorrhagic Escherichia coli (EHEC) is considered the most common cause. The majority of outbreaks and sporadic cases in humans have been associated with serotype O157:H7, although other O:H serotypes have been isolated, and they are a subgroup of Verocytotoxin-producing Escherichia coli (VTEC). Cattle are the principal reservoir of VTEC. Infections in humans are a consequence of consumption of undercooked meat, raw milk and other contaminated food or water. Direct contact with animals or people infected is another source of infection. PMID- 15338983 TI - [Argentine Consensus of Respiratory Rehabilitation]. AB - A group of pulmonologists and physical therapists from the Asociacion Argentina de Medicina Respiratoria revised the scientific literature on Respiratory Rehabilitation (RR) to elaborate evidence-based national recommendations to promote its use. RR is a multidisciplinary program of care for patients with chronic respiratory impairment, individually tailored, designed to optimize physical and social performance and autonomy of patients. It is particularly indicated in patients with Chronic Obstructive Pulmonary Disease (COPD) with exercise intolerance. Inclusion and exclusion criteria, guidelines for initial evaluation and follow up have been defined. The resources needed were defined. It was recommended a hospital ambulatory program with domiciliary complement. A pulmonologist and physical therapist were required for the program as minimum. Aerobic training was recommended for lower limb (LL) (Evidence A) and upper limb (UL) (Evidence B), strength training for LL and UL (Evidence C), as well as respiratory muscles training by resistive inspiratory threshold load (Evidence D) and other physiotherapy techniques were recommended for specific patients. In addition recommendations have been made for educational objectives of the program, nutritional and psychological support. The positive impact of RR on health care was analyzed through the reduction in exacerbation of COPD, length of hospital stay and cost. RR is a key component in the treatment of COPD patients. This evidenced-based consensus statement was prepared to provide recommendations to be implemented nationally. PMID- 15338984 TI - [Clinical medicine and the laboratory. Another appreciation]. PMID- 15338985 TI - [The lynx, the microscope and the bees]. PMID- 15338986 TI - [Water channels and the Nobel Prize]. PMID- 15338987 TI - [Authorship of medical publications]. PMID- 15338988 TI - [Clinical medicine and the laboratory, before and now. An appreciation]. PMID- 15338989 TI - [Clinicopathological conferences]. PMID- 15338990 TI - IHF survey reveals progress on eHealth strategies. PMID- 15338991 TI - Recent reforms of the German health care system. PMID- 15338992 TI - Healing Europe's ailing health centres in an enlarged European Union. An interview with professor Per-Gunnar Svensson. Interview by Kellie Peakman. PMID- 15338993 TI - Market principles in health care and social security policy in Japan. AB - Although health care in Japan is under the management of an obligatory insurance system, it is within the framework of a capitalist economy, and has helped achieve longevity during the post-war period. However, average lifetime has been improving in western European and Asian countries that have developed later. It has also been said that higher longevity is not necessarily due only to health care but also to the enhancement of environmental health achieved by economic improvements. On the other hand, the so-called 'development' led by capitalism and the market economy, and the luxuries that sometimes can be construed as uncultured, have caused unnecessary environmental destruction and disparities in wealth. Is it too cynical to think that the extended lifespan of the advanced countries has been achieved at the expense of the epidemics, refugee problems and wars which have resulted in a reduced lifespan in developing countries? It is said that capitalism is an economic ideology that includes many contradictions and is following a path of destruction. In addition, under the name of globalization, capitalism has continuously and rapidly caused vicious cycles of corruption, which are features commonly seen in today's world. It is still common that financial failures and threats are indirectly solved by initiating wars--a method completely inimical to health care. Along with environmental factors and the logic of this market doctrine, we have been trying to reform our financially collapsed health care system. However, we cannot count on the 'durability' of any reform conducted without some awareness of our economic mistakes. It is often said that it is only because of the existence of the economy that we have health care. However, it is more realistic to say that stable health care and social security lead to a stable economy. Health care did not collapse. It was the market economy upon which health care depended that collapsed. Therefore, one must not consider that health care is subordinate to the economy. Without corrections in the failure of the market economy and in the 'trickle down' economic ideology, health care and social welfare cannot be established on a worldwide basis--the medical community should now take a leading position to emphasize this point. PMID- 15338994 TI - Challenges confronting the health workforce in sub-Saharan Africa. AB - Sub-Saharan Africa and the international health community face a daunting challenge to deal with an extraordinary disease burden and improve the health status of Africans. Despite decades of effort to provide effective, equitable and affordable health care services, the health indices of Africans have stagnated and in some instances have deteriorated. Africa is the only continent that has not fully benefited from recent advances in biomedical sciences that brought health tools and technologies to tackle most of the disease burden. The emergence of the HIV/AIDS epidemic has confounded the health scene and posed further challenges. Several factors are responsible for this state of affairs: macro factors, that represent the broader socio-cultural environment that impact on health, and micro factors, which are largely health sector specific. There is increasing recognition that the major limiting factor to improved health outcomes is not lack of financial resources or health technologies but the lack of implementation capacity which depends on the presence of a functional health system. The drivers and architects of this are health workers, 'the most important of the health system's input'. The Commission on Macroeconomics and Health advocates a greatly increased investment in health rising in low income countries to a per capita expenditure of US $34 per year and states that the problem in implementing this recommendation is not difficulty in raising funds but the capacity of the health sector itself to absorb the increased flow. Yet, until fairly recently sufficient attention has not been directed to the role of the health workforce. The failure to develop and deploy an appropriate and motivated health workforce, and the environment necessary for the workforce to perform optimally is clearly a critical determinant of the health status of Africans. This paper summarizes key issues facing the workforce and outlines a framework to develop strategies to address them. PMID- 15338995 TI - The future of hospital-based radiological imaging. AB - Radiologic imaging has made dramatic progress over the last decades. The authors analyse all areas of current radiological imaging including conventional radiology, ultrasonography, computed tomography, magnetic resonance imaging, interventional radiology and nuclear medicine. Future opportunities and challenges regarding the subsequent development of imaging in general and radiology in particular are highlighted. Thus, the authors seek answers to questions such as: Will there be a need for radiologists? Will imaging become too costly? Possible remedies are suggested. PMID- 15338996 TI - Making hospitals safer: the need for a global strategy for infection control in health care settings. AB - Healthcare Associated Infections (HAI) have a substantial impact on morbidity, mortality and health costs. At least 5 to 10% of patients admitted to acute care hospitals acquire an infection. Factors that promote HAI include underlying diseases and decreased patient immunity; invasive diagnostic and therapeutic techniques, the widespread antimicrobial resistance, lack of infection control measures and environmental hygiene. HAI are preventable and infection control programmes are cost-effective. Health care workers education, HAI surveillance, appropriate legislation and basic infection control measures are the essential elements of an infection control programme. A recent consultation on the 'Prevention and Control of HAI' identified the need for an international strategy that would establish standards, procedures and methods for HAI surveillance, prevention and control and promote their implementation at national level. PMID- 15338997 TI - A European social model: the way forward in health care. PMID- 15338998 TI - Male and female fertility and hatchability in chickens: a longitudinal mixed model approach. AB - The objective of the current study was to investigate different approaches for handling missing records and to develop and implement a multivariate longitudinal mixed model for the genetic evaluation of male and female fertility and hatchability in chickens. The traits recorded on a weekly basis were eggs set (E), percentage fertility (F), and percentage hatch of fertile eggs (H). Three approaches for handling missing records were investigated: 1) all records with zero weekly laid eggs were removed and remaining records with missing F and H were predicted (M1); 2) missing records, including zero weekly laid eggs, were assumed known and equal to zero (M2); and 3) zero weekly laid eggs were assumed as a valid record and missing F and H were predicted (M3). A longitudinal mixed model was used for the multiple trait analysis of E, F, and H. Fixed effects included week-flock, age of service sire (rooster in the pen at data collection), and age of hen. Unrelated service sire, additive hen, permanent environmental, and the residual term were included as random effects. Heritability estimates ranged from 0.06 to 0.13 for E, F, and H. Heritability estimates of F and H were overestimated using M2. Correlations among the 3 traits were highest using M2 and lowest using M1. Pearson correlations indicated that reranking of birds and service sires could occur when using M1. Therefore, M1 should not be used for the analysis of longitudinal data of fertility and hatchability in chickens. Furthermore, M3 seems to be the most ideal method for handling missing records. PMID- 15338999 TI - Components of feed efficiency in broiler breeding stock: influence of water intake and gastrointestinal contents. AB - An experiment was conducted with 3 lines of broilers to evaluate the potential roles of water intake and fill of the gastrointestinal (GI) tract on feed conversion ratio (FCR) as measured in a 1-wk FCR test. Birds were reared to juvenile selection age (41 d) and subsequently selected, with the best 18 to 29% of male chicks placed into cages for FCR testing. Feed and water intake were highly correlated in all lines, with each in turn highly correlated with weight gain on test. Although filling of the GI tract differed between the top and bottom halves of the FCR distribution, FCR as measured and corrected for weight of GI tract contents were highly correlated (r = 0.59 to 0.93). Differences between FCR classes were striking for weight gain, but differences between FCR class groups were subtle for starting weight and feed intake. These subtle differences are difficult to detect in a single generation, but may add up through generations of selection. Based on the results of the current and previous studies, it appears that selection for improved FCR works at multiple levels to improve efficiency of feed conversion in broiler breeding stock. PMID- 15339000 TI - Optimization of phase feeding of starter, grower, and finisher diets for male broilers by mixture experimental design: forty-eight-day production period. AB - A mixture experiment, a variant of response surface methodology, was designed to determine the proportion of time to feed broiler starter (23% protein), grower (20% protein), and finisher (18% protein) diets to optimize production and processing variables based on a total production time of 48 d. Mixture designs are useful for proportion problems where the components of the experiment (i.e., length of time the diets were fed) add up to a unity (48 d). The experiment was conducted with day-old male Ross x Ross broiler chicks. The birds were placed 50 birds per pen in each of 60 pens. The experimental design was a 10-point augmented simplex-centroid (ASC) design with 6 replicates of each point. Each design point represented the portion(s) of the 48 d that each of the diets was fed. Formulation of the diets was based on NRC standards. At 49 d, each pen of birds was evaluated for production data including BW, feed conversion, and cost of feed consumed. Then, 6 birds were randomly selected from each pen for processing data. Processing variables included live weight, hot carcass weight, dressing percentage, fat pad percentage, and breast yield (pectoralis major and pectoralis minor weights). Production and processing data were fit to simplex regression models. Model terms determined not to be significant (P > 0.05) were removed. The models were found to be statistically adequate for analysis of the response surfaces. A compromise solution was calculated based on optimal constraints designated for the production and processing data. The results indicated that broilers fed a starter and finisher diet for 30 and 18 d, respectively, would meet the production and processing constraints. Trace plots showed that the production and processing variables were not very sensitive to the grower diet. PMID- 15339001 TI - Comparison of zinc acetate and propionate addition on gastrointestinal tract fermentation and susceptibility of laying hens to Salmonella enteritidis during forced molt. AB - Feed deprivation is the most common method used to induce molting and stimulate multiple egg-laying cycles in laying hens for commercial egg production. Unfortunately, an increased risk of Salmonella enteritidis (SE) colonization may result from the use of this method. Methods to stimulate multiple egg-laying cycles without increasing the risk of SE are needed. In each of 3 experiments, hens over 50 wk of age were divided into groups of 12 and placed in individual laying cages. One week before dietary changes, hens were put on an 8L:16D photoperiod that continued for the 9-d experimental period. Hens in all treatments were challenged orally with 10(4) cfu of SE on the fourth day. Treatments were full fed hens (nonmolted, NM), nonfed hens (molted, M), a zinc acetate diet (ZAC), and a zinc propionate diet (ZPR). The zinc diets contained 10,000 mg of zinc per kilogram of diet. Body weight losses were significantly higher in the M, ZPR, and ZAC treatments than in the NM treatment. Crop lactic acid decreased more in M, ZPR, and ZAC treatments than in NM hens in trial 2. Crop pH was significantly (P < 0.05) lower in NM hens than in M, ZAC, and ZPR hens in trial 2. Although cecal individual or total volatile fatty acids (VFA), and lactic acid were not significantly (P > 0.05) different between NM hens and M, ZAC and ZPR hens in trial 1, lactic acid was significantly (P < 0.05) higher in NM hens than in M, ZAC and ZPR hens (trial 2), and cecal total VFA were lower in M hens than in NM, ZAC and ZPR hens (trial 3). Colonization of SE in the crop and ceca was higher in the M and ZPR hens (trials 1 and 2). Liver, spleen, or ovary invasion by SE was higher in the M and ZPR hens (trials 1 and 2) than in NM hens. At the zinc concentration used in these studies, the zinc dietary regimens may be effective for reducing the risk of SE during induced molt. PMID- 15339002 TI - Toxicity of maize containing known levels of fumonisin B1 during force-feeding of ducks. AB - The toxicity of maize containing known doses of fumonisin B1 (FB1) was investigated in mallard ducks during force-feeding. Seventy-five ducks at 12 wk of age were randomly divided into 3 groups of 25, and received control maize, naturally contaminated maize containing 20 mg/kg of FB1, or a mixture of control and contaminated maize (50/50, vol/vol). Force-feeding was performed during 12 d that correspond to a final average feed intake of approximately 10 kg of maize per duck. At the end of the study, 8% mortality was observed in ducks fed 20 mg of FB1/kg of feed, whereas no mortality occurred in the other groups. Liver weight, and plasma concentrations of protein, cholesterol, alanine aminotransferase (ALAT), and lactate dehydrogenase (LDH) were increased by force feeding, whereas feed conversion ratio appeared decreased by the toxin. Microscopic examination of the liver showed that steatosis was mostly macrovacuolar in control ducks, whereas it was microvacuolar in ducks fed 20 mg of FB1/kg of feed. Free sphingolipid concentrations were measured in liver and plasma. Sphinganine (Sa) and sphinganine to sphingosine (Sa/So) ratio were increased in all treatment groups. These parameters were not affected by force feeding and all individual values obtained in the treated ducks were higher than those obtained in control ducks. Our results suggest that free Sa level and Sa/So ratio can be used to reveal exposure of ducks to FB1 at doses of 10 mg/kg or greater in feed. PMID- 15339003 TI - On-farm euthanasia of broiler chickens: effects of different gas mixtures on behavior and brain activity. AB - The purpose of this study was to investigate the suitability of gas mixtures for euthanasia of groups of broilers in their housing by increasing the percentage of CO2. The suitability was assessed by the level of discomfort before loss of consciousness, and the killing rate. The gas mixtures injected into the housing were 1) 100% CO2, 2) 50% N2 + 50% CO2, and 3) 30% O2 + 40% CO2 + 30% N2, followed by 100% CO2. At 2 and 6 wk of age, groups of 20 broiler chickens per trial were exposed to increasing CO2 percentages due to the injection of these gas mixtures. Behavior and killing rate were examined. At the same time, 2 broilers per trial equipped with brain electrodes were observed for behavior and brain activity. Ten percent of the 2-wk-old broilers survived the increasing CO2 percentage due to the injection of 30% O2 + 40% CO2 + 30% N2 mixture, therefore this mixture was excluded for further testing at 6 wk of age. At 6 wk of age, 30% of the broilers survived in the 50% N2 + 50% CO2 group. The highest level of CO2 in the breathing air (42%) was reached by the injection of the 100% CO2 mixture, vs. 25% for the other 2 mixtures. In all 3 gas mixtures, head shaking, gasping, and convulsions were observed before loss of posture. Loss of posture and suppression of electrical activity of the brain (n = 7) occurred almost simultaneously. The results of this experiment indicate that euthanasia of groups of 2- and 6-wk-old broilers by gradually increasing the percentage of CO2 in the breathing air up to 40% is possible. PMID- 15339004 TI - Efficacy of Leuconostoc mesenteroides (ATCC 13146) isomaltooligosaccharides as a poultry prebiotic. AB - The complex dietary carbohydrates, called prebiotics, have been used to control Salmonella and improve intestinal bacterial balance in broilers. Leuconostoc mesenteroides (ATCC 13146) isomaltooligosaccharides (IMO) stimulate growth of Bifidobacterium and Lactobacillus and are not used by Salmonella or Escherichia coli. We tested the efficacy of these IMO as a prebiotic. IMO, compared with fructooligosaccharides (FOS) as sole carbon source, promoted growth of chicken cecal isolates and Bifidobacterium. Cecal isolates and Salmonella typhimurium grown in mixed culture on IMO reduced the Salmonella population. Cecal isolates grown on IMO showed higher viable counts and faster growth than Salmonella, indicating a potential value for these oligomers for poultry intestinal microflora modification. PMID- 15339005 TI - Responses of growing broilers to diets with increased sulfur amino acids to lysine ratios at two dietary protein levels. AB - An experiment with 1,440 male Cobb 500 and 1,440 male Ross 308 broilers (14 to 35 d of age) was conducted to investigate the effects of diets having 4 levels of digestible methionine plus cysteine (SAA) on various performance criteria at 2 dietary protein levels (20.5 and 26.0%). Two corn-soybean meal/poultry by-product basal diets were formulated to contain 3,060 kcal/kg MEn and either 20.5 or 26.0% balanced protein, and 1.12 and 1.46% digestible (according to table values) lysine, respectively. Except for SAA, the ratios between essential amino acids were kept identical in both diets according to the ideal protein concept. The ratio between digestible SAA and digestible Lys was 50%. All remaining nutrients met or exceeded NRC (1994) recommendations. Graded levels of SAA were supplemented to obtain digestible SAA to Lys ratios of 62, 69, and 77%, with 77% representing an optimized amino acid balance. Increasing the protein level clearly improved weight gain, feed conversion, breast meat yield, and abdominal fat content. Increasing SAA levels resulted in strong nonlinear or linear dose responses at both protein levels and for both strains. Regression analysis suggested that reducing digestible SAA in a balanced protein (diets with SAA:Lys of 77%) impairs performance, and that optimum SAA:Lys ratio for growing broilers might be higher than 77%, although ANOVA revealed no significant improvement with an SAA:Lys ratio higher than 69%. Responses provide evidence that optimum dietary SAA level depends on dietary protein level and should therefore be related to the protein content. PMID- 15339006 TI - Response to lysine intake in composition of body weight gain and efficiency of lysine utilization of growing male chickens from two genotypes. AB - Male chickens of a broiler (B) and a layer (L) genotype were grown in floor pens from d 8 to 21 posthatch in groups of 10. Three pens per genotype were allocated to each of 10 experimental diets. The diets were offered ad libitum and they differed in lysine concentration from 3.8 to 16.8 g/kg. The source of supplemental lysine was L-lysine x HCl. All birds were killed at the end of the experiment, and representative birds (3 groups of 10 per genotype) were killed at the start for baseline measurements. Accretions of protein, fat, energy, and amino acids were determined by comparative body analysis. Responses were described with sigmoidal and exponential functions. Additionally, the net disappearance rate (NDR) of amino acids from the small intestine was studied with the basal diet (3.8 g of lysine/kg) using 6 replicated pens of 15 birds per genotype. Titanium dioxide was the indigestible marker. Net disappearance rates were not significantly different between genotypes for CP or any amino acid. Responses to incremental lysine concentration were nonlinear for both genotypes but distinctly different in magnitude between genotypes. Estimated y(max) values for 14-d BW, protein gain, and gain/ feed ratio were 534 (B) and 153 (L) g, 87.1 (B) and 28.7 (L) g, and 0.82 (B) and 0.71 (L) g/g. Protein accretion approached 95% of the estimated y(max) with dietary lysine concentrations of 12.5 (B) and 10.4 (L) g/kg. The amino acid profile of accreted whole body protein was different between genotypes, and was affected by supplementary lysine. Lysine content in accreted whole body protein approached upper values of 7.4 (B) and 5.6 (L) g/16 g of N with increasing dietary lysine concentration. Marginal efficiency of lysine utilization, determined as delta lysine accretion/delta lysine intake, showed maxima of 99% (B) and 74% (L). These maxima were achieved at intakes which were much lower than those needed for high protein accretion. It was concluded that the efficiency of amino acid utilization may depend on genotype, perhaps due to differences in the relative proportion of different protein fractions to whole body protein and due to differences in the ratio of synthesis and degradation of body proteins. Nonlinear relationships and different amino acid pattern of accreted body protein should be implemented in future models of requirements. PMID- 15339007 TI - Evaluation of broiler performance when fed Roundup-Ready wheat (event MON 71800), control, and commercial wheat varieties. AB - We evaluated the nutritional value of broiler diets containing approximately 40% wheat grain from Roundup Ready wheat (MON 71800), its similar nontransgenic control (MON 71900), or reference commercial wheat varieties. The feeding trial lasted 40 d, and each treatment consisted of 10 replicates of 1-d-old Ross 308 broilers (5 pens of males and 5 pens of females). Each pen contained 12 birds, and at d 13 birds were randomly removed until 9 birds remained. Body weight and feed intake were measured on pen basis at 40 d. At d 41, four broilers per pen were slaughtered. The carcasses were dissected, and cut-up yields were determined. Dry matter, protein, and fat contents of breast meat were determined. The data were analyzed by an ANOVA procedure. The BW and feed conversion at d 40 averaged 2,450 g and 1.52, respectively. There were no significant treatment x sex interactions, except for evisceration yield with significant differences (P < 0.05) in yield between birds fed 2 commercial wheat varieties. Data for final BW, feed conversion, carcass yield, and breast meat were not statistically different (P < 0.05) between broilers fed MON 71800 or MON 71900 or the population of birds fed commercial wheat varieties, except a lower carcass yield at d 41 for birds fed the nontransgenic control wheat. Thus MON 71800 was nutritionally equivalent to nongenetically modified wheat varieties when fed to broilers. PMID- 15339008 TI - The effects of copper on the efficacy of phytase, growth, and phosphorus retention in broiler chicks. AB - Copper is often added to broiler diets at prophylactic concentrations as an antimicrobial despite purported chelation with and reduced utilization of phytin phosphorus. Therefore, male chicks were fed 0, 62.5, 125, 250, or 375 ppm Cu from Cu sulfate in combination with 600 phytase units (FTU)/kg phytase from 9 to 22 d of age (6 cages/diet, 8 birds/cage). Nonphytate phosphorus (NPP) and Ca were formulated to 0.2 and 0.7% of the diet, respectively. Three additional control diets were formulated to contain 0.27, 0.34, and 0.40% NPP, each with 0.7% Ca. Birds fed increasing concentrations of Cu with 600 FTU phytase/kg had linear reductions in performance characteristics (P < or = 0.05). Birds fed increasing concentrations of Cu with 600 FTU phytase/kg had linear increases in toe ash percentage (P < or = 0.027), but tibia ash percentage was not affected (P > 0.05). Birds fed increasing Cu concentrations with 600 FTU phytase/kg had linear reductions in apparent P retention as a percentage of total P (P < or = 0.0005). Supplementation with increasing concentrations of Cu to a diet containing 600 FTU phytase/kg resulted in decreases in 21 d BW, BW gain, feed consumption, feed conversion, tibia and toe ash weights, and apparent P retention as a percentage of total P. In this experiment, Cu supplementation did not reduce the efficacy of phytase (i.e., improvement in apparent P retention with phytase supplementation) but did decrease apparent P retention, BW gain, feed consumption, feed conversion, and tibia ash and toe ash weights. PMID- 15339009 TI - Expression of the chicken peroxisome proliferator-activated receptor-gamma gene is influenced by aging, nutrition, and agonist administration. AB - Peroxisome proliferatior-activated receptor-gamma (PPARgamma) is a transcription factor that modulates lipid and glucose metabolism in mammals. The aim of the present study was to investigate whether chicken PPARgamma is expressed in tissues in a similar manner to mammalian PPAR and whether it is involved in the regulation of lipid metabolism, particularly in the regulation of fat accumulation in adipose tissue and ovaries. In 30-wk-old chickens, PPARgamma mRNA was detected in most tissues that were examined. Of those tissues expressing chicken PPARgamma mRNA, the lowest expression levels were found in adipose tissue, the tissue that in mammals was shown to express the highest levels of PPARgamma mRNA. Chicken PPARgamma mRNA expression in abdominal adipose tissue tended to increase with age, as shown by higher expression levels at 6 wk than at 1 and 2 wk of age. With regard to nutritional modulation, PPARgamma mRNA levels in abdominal adipose tissue were significantly higher in broiler chickens fed for 7 d a diet containing 8% safflower oil (18:2-rich) or linseed oil (18:3-rich) compared with chickens fed a diet containing olive oil (18:1-rich). In contrast, feeding a 3% cholesterol-supplemented diet for 7 d resulted in no changes to adipose PPARgamma mRNA expression. In broiler chickens orally administered troglitazone, a PPARgamma ligand, abdominal fat pad weight and PPARgamma and lipoprotein lipase (LPL) mRNA levels were significantly increased relative to those of control chickens. Levels of PPARgamma mRNA in liver, skeletal muscle, and ovaries were increased with the onset of egg laying, whereas in adipose tissue the level of PPARgamma mRNA was decreased. These findings suggest that PPARgamma plays an important role in the regulation of fat deposition and egg production and the characteristic pattern of PPARgamma mRNA expression may be indicative of specific differences in the lipid and glucose metabolism of chickens compared with mammals. PMID- 15339010 TI - Validation of dual-energy x-ray absorptiometry for determining in vivo body composition of chickens. AB - The purpose of this study was to validate dual-energy x-ray absorptiometry (DXA) for measuring the body composition of chickens in vivo. Four trials were conducted with broiler chickens using a DXA instrument (Lunar, DPX-L) and small animal total body scan software (version 4.7a). In the first 2 trials, the effects of scan mode (high resolution or detail slow), scanning position (ventral or dorsal), and the use of attenuating materials (AM) (2-mm polyvinyl chloride or 4-mm polystyrene) on the precision and values of the DXA parameters body mass, lean tissue mass, fat tissue mass and percentage, bone mineral content (BMC), and bone mineral density (BMD) were evaluated. The precision was highest for body mass and lean tissue mass, followed by BMC and BMD, and was lowest for fat tissue mass and percentage. The precision of the measurements was not influenced by scan mode, position, or type of AM. In contrast, the values for all DXA parameters except body mass were significantly influenced by the scan mode but not by the position. The high resolution mode gave significantly higher estimates of fat mass and BMC but significantly lower measures of lean tissue mass and BMD compared with the detail slow mode. A significant difference between AM was only observed for the DXA estimates of fat tissue mass and fat percentage. In trial 3, the accuracy of the DXA measurements was tested by comparison with chemical body composition analysis. Linear regression equations between the respective DXA and chemical parameters were established. High correlations (r > 0.9; P < 0.0001) were obtained for all parameters, except for fat percentage (r = 0.593; P < 0.05). The purpose of the validation trial was to compare the predicted body composition based on the DXA measurements with established equations and the chemical body composition. There was extremely good agreement for body mass, lean tissue mass, and fat tissue mass and percentage, but not for ash weight. It is concluded that, after proper methodological standardization and application of specifically determined regression equations, DXA can be used for estimating the body composition of chickens in vivo. However, the regression equations are strictly limited to one particular instrument, software version, and applied methodology. PMID- 15339011 TI - Influence of dietary calcium and phytase on phytate phosphorus hydrolysis in broiler chickens. AB - The effect of Ca and phytase on phytate phosphorus (PP) hydrolysis was studied in vitro and in vivo. In vitro, PP hydrolysis by a 3-phytase and a 6-phytase was studied at pH 2.5 and 6.5 with Ca added at levels equivalent to 0, 0.1, 0.2, 0.4, 0.7, or 0.9% of the diet. Irrespective of enzyme, Ca at a level as low as 0.1% reduced (P < 0.05) PP hydrolysis at pH 6.5. To test these effects in vivo, 22-d old male broilers were fed 1 of 6 diets (10 replicate pens of 4 birds per diet) for 30 h. The experimental design was a 3 x 2 factorial arrangement of 3 phytase treatments (0, 500 U of phytase A/kg of diet, and 500 U of phytase B/kg of diet) and 2 added Ca levels (0 and 0.5% from CaCO3) to a corn-soy basal diet. Adding Ca to the diet resulted in a reduction (P < 0.05) in ileal PP disappearance from 69.2 to 25.4% when the 0 and 0.5% added Ca diets were fed, respectively, and in apparent ileal Ca and P absorption (46.3 to 33.6% and 67.9 to 29.4% when 0 and 0.5% Ca were added, respectively). Inclusion of a 3-phytase improved (P < 0.05) ileal PP disappearance from 25.4 to 58.9% in diets containing 0 and 0.5% added Ca, but the improvement was less pronounced with a 6-phytase. Apparent ileal Ca absorption was improved (P < 0.05) when Ca, phytase, or both were added to the diet. PMID- 15339012 TI - Threonine needs of broiler chickens with different growth rates. AB - The Thr needs in 3 commercial broiler strains (A, multipurpose; B, high yield; C, high yield) known to differ in terms of feed intake, growth rate, and breast yield were evaluated. Birds were randomized across 96 floor pens (12 birds/pen), received a common diet from d 1 to 20, and were fed graduations of Thr (0.52 to 0.87% total Thr in 0.07% increments) from d 21 to 42. Treatments (3 x 6 factorial) were replicated 5 or 6 times. The corn, soybean meal, and peanut meal test diet contained 0.43 and 0.96% digestible Thr and Lys, respectively. An additional group of strain C birds (6 pens) was maintained on a corn-soybean meal diet containing surfeit Thr (0.73% of diet). Birds fed the corn and soybean meal diet performed similarly (P < or = 0.05) to birds fed peanut meal diets. A feed conversion interaction (P < or = 0.05) occurred indicating that strain C was more sensitive to Thr deficiency than strains A and B. The abdominal fat interaction (P < or = 0.05) indicated that strain A had more relative abdominal fat than strains B and C. All strains differed (P < or = 0.05) in terms of BW gain (A, 78.2; B, 75.1; C, 72.9 g/d). Strain C had the lowest (P < or = 0.05) feed intake, which resulted in the lowest (P < 0.05) Thr intake, but it had the highest (P < or = 0.05) breast meat yield. Most parameters tested yielded quadratic (P < or = 0.05) models whereby Thr estimates could be predicted. Namely, BW gain and breast meat yield resulted in total Thr estimates (95% of maximum response) of 0.74 and 0.71%, respectively, which are in close agreement with the 1994 NRC (0.74%). The plasma Thr sigmoid response verified the former estimates. Analysis of strain intercepts and slopes as affected by Thr differed (P < or = 0.05) in terms of feed intake but not BW gain or breast meat yield. The 21 to 42 d Thr need across strains was estimated as 0.74% total or 0.65% digestible. Because dietary Lys was not in excess of the bird's needs, the former digestibility estimate equated to a Thr/Lys of 0.68. PMID- 15339013 TI - Tracing metabolic routes of feed ingredients in tissues of broiler chickens using stable isotopes. AB - The present study aimed to quantify the proportion of 13C from energy and protein feed ingredients that follow the metabolic routing of the liver and muscle in broiler chickens. A stable isotope of carbon technique was used that is based on the isotopic discrimination that occurs in the plants during the photosynthesis process. One-day-old male chicks were subjected to treatments based on free choice of energy and protein sources. Rice bran (R) and soybean meal (S), C3 plants, have higher isotopic ratios than corn (C), a C4 plant, and corn gluten meal (G). Choices were R+S, C+G, R+G, C+S, or R+C+G+S. A complete feed (CF) was a sixth treatment. Feed intake and BW were measured at 30 d of age, when liver and breast muscle were collected for isotopic analysis. Treatments affected the amount of feed intake and the choices of energy or protein sources. Complete feed had the largest intake, differing from the other treatments that had free-choice feeding. Final BW was a direct reflection of consumption by these birds in all treatments. The isotopic results indicated that the 13C/12C ratio was generally higher in breast muscle than in liver, probably because of higher protein content. Moreover, in the liver, the proportion of 13C retained from the energy ingredient was greater than the proportion from the protein ingredient. That is in contrast to muscle, where the proportion of 13C retained from the protein ingredient was greater than from the energy ingredient that was self-selected. PMID- 15339014 TI - Dietary glycine needs of broiler chicks. AB - Dietary Gly might become a limiting factor in all-vegetable diets fed to broiler chicks when low CP is formulated in combination with marginal levels of dietary Thr and Ser. A study was conducted to evaluate dietary Gly needs of broiler chicks. Day-old Ross 508 male chicks were placed in 32 floor pens (15 chicks/pen). Chicks were fed a common prestarter diet from 0 to 7 d of age and then fed a diet that contained progressive amounts of dietary Gly ranging from 0.62 to 1.22% from 7 to 20 d of age. Treatment effects were observed for weight gain and feed conversion. Chicks responded in a quadratic manner to supplementation with dietary Gly. The dietary Gly level necessary to support maximum growth and feed conversion for the chick from 7 to 20 d of age was estimated to be at 0.98 (1.76% Gly + Ser) and 1.02% (1.80% Gly + Ser), respectively. Plasma Thr and Ser were unaffected by Gly supplementation, but plasma free Gly increased linearly. Dietary Gly may need to be considered as a limiting nutrient in early nutrition, especially if CP is low, and only vegetable ingredients are being used. PMID- 15339016 TI - Determination of mitochondrial function and site-specific defects in electron transport in duodenal mitochondria in broilers with low and high feed efficiency. AB - Duodenal mitochondria were isolated from broiler breeder males with high (0.79+/ 0.01, n = 9) and low (0.63+/-0.02, n = 9) feed efficiency (FE) to assess relationships of FE with duodenal mitochondrial function and site-specific defects in electron transport. Sequential additions of adenosine diphosphate (ADP) resulted in 1) higher respiratory control ratio (RCR; an index of respiratory chain coupling) in high FE mitochondria provided succinate, and 2) higher ADP to oxygen ratio (ADP:O; an index of oxidative phosphorylation) in low FE mitochondria provided NADH-linked substrates (malate, pyruvate, or both). Basal electron leak, measured as H2O2 production, was greater in low FE mitochondria provided succinate (P = 0.08) or NADH-linked substrates. As H2O2 levels were elevated in low FE compared with high FE mitochondria by complex I (P+/-0.07) and complex II inhibition, the higher basal electron leak in low FE mitochondria was apparently due to site-specific defects in electron transport at complexes I and II. Elevations in H2O2 above basal levels indicated that high FE mitochondria may also exhibit electron transport defects at complexes I and III. Despite an ability to produce adenosine triphosphate (ATP) that was equal or superior to that demonstrated in high FE duodenal mitochondria, low FE mitochondria exhibited a greater inherent degree of electron leak. The results provide insight into the role that duodenal mitochondria play in the phenotypic expression of FE in broilers. PMID- 15339015 TI - Activity of phospholipases A and lysophospholipase in turkey semen and oviducal fluid. AB - Changes in lipid composition of turkey semen have previously been reported to occur during in vitro storage and may be mediated by endogenous hydrolysis of phospholipids. To investigate the presence of phospholipases able to initiate such degradation, phospholipaseA2 (PLA2), phospholipase A1 (PLA1), and lysophospholipase (LPLase) activities were measured in turkey spermatozoa and seminal plasma. These enzymes were also measured in the oviductal fluid because they may be involved in the process prior to fertilization in the female. In spermatozoa and seminal plasma, the major PLA2 was a calcium-dependent and sodium deoxycholate (DOC) stimulated enzyme. However, calcium-independent PLA2 activities were also detected with different characteristics in spermatozoa (DOC inhibited enzyme) and seminal plasma (DOC stimulated enzyme). Additionally, PLA1 activity and high LPLase activity were present in spermatozoa and seminal plasma. In vitro storage of semen for 48 h did not affect PLA2 and LPLase activities. By contrast, PLA1 was the major phospholipase activity detected in oviductal fluid. A PLA2 activity stimulated by calcium or DOC and LPLase activity were also detected, but both were low relative to PLA1. These results showed that turkey semen had several enzymatic activities able to hydrolyze phospholipids. In addition, the phospholipase activities described here in the oviductal fluid could be involved in membrane destabilization prior to fertilization. PMID- 15339017 TI - Comparison of chicken genotypes: myofiber number in pectoralis muscle and myostatin ontogeny. AB - This study was performed to evaluate breast muscle development in chicken genotypes divergently selected for muscularity. In the first experiment, 2 commercial broiler lines (a high breast yield, HBY, and a normal breast yield broiler strain-cross, NBY) and a Leghorn line were grown up to 35 d to evaluate BW, breast weight, and breast yield. At 7 and 21 d of age, pectoralis muscle was used to estimate myofiber density (MFD, number of myofibers per mm2) and total apparent myofiber number (MFN). In the second experiment, the ontogeny of myostatin was determined from broiler- and Leghorn-type chick embryos, at embryonic days 1 to 20 (E1 to E20), using reverse transcription (RT)-PCR. As expected, the Leghorn line had lower BW, breast weight, and breast yield than broiler lines. The HBY line showed higher breast yield at all ages evaluated, but lower BW at 21 and 35 d than the NBY line. The Leghorn line had 45% higher MFD than broilers, which indicates an increased cross-sectional area of the myofibers in broiler lines. No MFD difference was observed between the broiler strains (P > 0.05). The myofiber number of broilers was more than twice that of Leghorns and HBY had 10% higher MFN than the NBY line. Myofiber number was correlated to BW (r = 0.58), breast weight (r = 0.58), and breast yield (r = 0.69). Conversely, MFD showed negative correlation with BW, breast weight, and breast yield (r = -0.85, 0.83, and -0.88, respectively). No effect of genotype or interaction between genotype and embryonic age was observed for myostatin expression. This study showed that broilers have higher MFN in the breast muscles than Leghorn-type chickens, and that high breast yield of broiler strains may be due to increased MFN. Higher muscularity of broilers, as compared with Leghorns, was not attributed to lower expression of myostatin during embryonic development. PMID- 15339018 TI - Monochromatic light stimuli during embryogenesis enhance embryo development and posthatch growth. AB - Photostimulation with green light accelerated BW and muscle development of broilers. In experiment 1, temperature sensors were inserted into 50 broiler eggs. The eggs were placed under 5 green light-emitting diode (LED) lamps at an intensity of 0.1 W/m2 at eggshell level for 5, 10, 15, 20, and 25 min (n = 10). Egg temperatures were recorded continuously. A high correlation was found between lighting period and egg temperature elevation, and an intermittent light regimen of 15 min on and 15 min off was found to eliminate light-induced egg overheating. In experiment 2, the effect of in ovo green light photostimulation on embryonic development was studied. Five hundred fertile eggs were divided into 2 groups: the first was photostimulated with green light from 5 d of incubation until hatch (0.1 W/m2 intensity) and the second was incubated in the dark. In ovo green light photostimulation caused a significant elevation in BW and breast muscle weight during embryo development and posthatch until 6 d of age. In experiment 3, 240 fertile broiler eggs were divided into 2 groups as described in experiment 2. At hatch, chicks from each in ovo light treatment were divided into 2 subgroups: the first was reared under green light and the second under white light. In ovo photostimulation with green light enhanced BW and breast muscle weight. However, rearing under green light did not have any synergistic effect on BW. Collectively, the results suggest that stimulation with green light enhances development and growth in chicks and that the best effect is achieved when this stimulus is provided during incubation. PMID- 15339019 TI - Differential expression of mitochondrial electron transport chain proteins in cardiac tissues of broilers from pulmonary hypertension syndrome-resistant and susceptible lines. AB - Pulmonary hypertension syndrome (PHS) is a metabolic disease associated with the rapid growth rate of modern broilers. Broilers susceptible to PHS experience sustained elevation of pulmonary arterial pressure leading to right ventricular hypertrophy and ultimately heart failure. Previous studies have shown that mitochondrial function is defective in broilers with PHS; they use oxygen less efficiently than broilers without PHS. In this study mitochondrial electron transport chain (ETC) protein levels were compared in cardiac tissues from PHS resistant and susceptible line broilers using quantitative immunoblots. Seven of 9 anti-mammalian mitochondrial ETC protein antibodies tested exhibited cross species reactivity. Six ETC proteins were differentially expressed in the right ventricles of broilers raised under simulated high altitude conditions (2,900 m above sea level). Four ETC proteins were present at higher levels in resistant line birds without PHS than in resistant line birds with PHS or in susceptible line birds with or without PHS. One ETC protein was present at higher levels in broilers without PHS than in broilers with PHS in both lines, and one ETC protein was present at lower levels in susceptible line birds without PHS than in susceptible line birds with PHS or in resistant line birds with or without PHS. Interestingly, differential expression of mitochondrial ETC proteins was not observed in the right ventricles of broilers raised at local altitude (390 m above sea level) nor was it observed in the left ventricles of broilers exposed to simulated high altitude. These results suggest that higher levels of mitochondrial ETC proteins in right ventricle cardiac muscle may be correlated with resistance to PHS in broilers. PMID- 15339020 TI - Microbiological quality of broiler carcasses during processing in two slaughterhouses in Turkey. AB - The effect of processing procedures on the microbial quality and safety of broiler carcasses was investigated in 2 processing plants. Neck skin samples were taken from broilers at the main stages of processing and changes in total viable count (TVC) and in the counts of Coliforms, Enterobacteriaceae, and Staphylococci/Micrococci were monitored. Processing reduced TVC more than 2 log cycles for the 2 processing plants investigated. The counts of Coliform bacteria decreased from 5.35 to 3.99 log cfu/g, Enterobacteriaceae from 5.36 to 3.81 log cfu/g, and Staphylococci/Micrococci from 6.90 to 4.11 log cfu/g for the first processing plant. The counts of Coliforms, Enterobacteriaceae, and Staphylococci/Micrococci were reduced from 5.67 to 3.92 log cfu/g, from 5.75 to 3.91 log cfu/g, and from 6.85 to 3.94 log cfu/g, respectively, by processing in the second plant. Cross-contamination with Salmonella spp. during processing was observed and the incidences of Salmonella spp. on the carcasses increased from 40 to 60% and from 33.3 to 40% during processing at plants 1 and 2, respectively. The incidences of Salmonella spp. in all broilers (n = 90 carcasses for each plant) were 36.6 and 31.1% at plants 1 and 2, respectively. Although a high proportion of microorganisms were removed from carcasses during processing, dissemination of Salmonella spp. was found to be unavoidable. PMID- 15339021 TI - Refrigeration and freeze-thaw effects on broiler fillets having extreme L* values. AB - Eight hundred broiler males were grown to 56 d and processed under common terms while maintaining individual identity. Front halves were deboned 24 h postmortem (PM) to obtain breast fillets, and CIELAB light reflectance was immediately measured on the skin side of each fillet. Fillets were bagged and held at 4 degrees C for 24 h, and then 20 fillets exhibiting the darkest (47.3 to 57.5), lightest (71.1 to 76.4), and median (63.7 to 64.0) L* values were selected and trimmed to best define the pectoralis major. Remeasurement of light reflectance at 48 h PM revealed decreased L* values solely associated with fillets having the highest L* at 24 h. One-half of the fillets representing each category was frozen (4 d at -20 degrees C) and thawed (3 d at 4 degrees C). The L* value, after thawing, decreased from the 48 h PM value, which equivalently occurred in for all L* categories. Although 48 h PM fillets from each L* category were of similar weights, their lengths and widths increased with L* value. Exudate lost with thawing increased with L* value and paralleled decreases in length and width to equalize dimensions among sources. PMID- 15339022 TI - The use of halothane and succinylcholine to identify broilers prone to developing pale, soft, exudative meat. AB - Within the last several years, the poultry industry has seen a dramatic increase in the occurrence of pale, soft, and exudative (PSE) meat. This problem is known to be associated with a rapid decline in postmortem (PM) muscle pH, which results in inferior protein functionality similar to that found in PSE pork. Many factors such as seasonal changes have been known to influence the occurrence of PSE meat in poultry and swine. Halothane and succinylcholine have been used within the pork industry to identify animals susceptible to stress and prone to developing PSE meat. The mechanism for the triggering of the PSE gene in poultry has not been fully understood. Therefore, a study was conducted to determine the effectiveness of screening broilers with halothane to identify those prone to developing PSE meat. Succinylcholine was used before slaughter to serve as a triggering agent for the PSE condition. At 4 wk of age, broilers from 4 commercial strains (n = 1,000) were subjected to 3% halothane gas and classified as either halothane positive (HAL+) or negative (HAL-) based on muscle rigidity within the legs. Although halothane sensitivity varied slightly among the strains, approximately 14% of the birds overall were classified as HAL+. All HAL- birds (n = 163) and an equal number of HAL-birds (n = 163) in each strain were grown to market age (7 wk) and were commercially processed. At the time of processing, half of the HAL+ and HAL- birds were injected intravenously with succinylcholine and were slaughtered at 0.25 h postinjection. Pectoralis muscle samples were collected at 0.25, 2, 5, and 24 h PM for the evaluation of rigor development (muscle pH) and meat quality (L* value, moisture, drip loss, and cook loss). Halothane sensitivity had no effect on rigor development, muscle color, or water-holding capacity in the 4 broiler strains. Although birds exhibited reactions to the halothane gas, the halothane sensitivity, along with the use of succinylcholine, was not able to identify birds prone to developing PSE meat. PMID- 15339023 TI - Breed differences in the biochemical determinism of ultimate pH in breast muscles of broiler chickens--a key role of AMP deaminase? AB - The biochemical determinism of ultimate pH (pHu) was studied in the pectoralis muscle of broiler chickens. Thirty birds of 3 genetic types (a fast-growing standard (FG), a slow-growing French "Label Rouge" (SG), and a heavy line type (HL)) were kept under conventional breeding methods until the usual marketing age (6, 12, and 6 wk for FG, SG, and HL birds, respectively). The birds were divided into 3 different antemortem treatment groups: minimum stress, shackling for a longer time (2 min), and heat stress (exposure to 35 degrees C for 3.5 h and shackling for 2 min before stunning). The birds were slaughtered on the same day. The pHu differed (P < 0.001) among the 3 genetic types, ranking as follows: FG (5.95+/-0.01) > HL (5.85+/-0.02) > SG (5.73+/-0.02). In SG and HL birds, pHu was strongly correlated with muscle glycogen content at slaughter (r = -0.74 and 0.82; P < 0.01 respectively), whereas this correlation was weak in FG birds. Regardless of genetic type, neither buffering capacity nor lactate accumulation significantly contributed to pHu variations (P > 0.05). The activity of adenosine monophosphate deaminase (AMPd) was significantly higher in FG chickens (0.98+/ 0.31; P < 0.05) than in HL and SG birds (0.46+/-0.24 and 0.34+/-0.18, respectively). Significant correlations were found between AMPd activity, pHu, and glycolytic potential (GP) at slaughter (r = 0.34 and -.29; P < 0.01, respectively). Further research is needed to study in more detail the role of AMPd in the determinism of pHu, particularly in fast-growing broilers. PMID- 15339024 TI - Impact of feathers and feather follicles on broiler carcass bacteria. AB - Genetically featherless and feathered broiler siblings were used to test the contribution of feathers and feather follicles to the numbers of aerobic bacteria, Escherichia coli, and Campylobacter in whole-carcass rinse samples taken immediately after carcasses were defeathered for 30 or 60 s. Numbers of spoilage bacteria were counted after the same fully processed carcasses were stored for 1 wk at 2 degrees C. In each of 3 replications, twenty-eight 11-wk old, mixed-sex, genetically featherless or feathered broilers were processed in a laboratory processing facility. Immediately after individual defeathering in a mechanical picker, carcasses were sampled using a carcass rinse technique. Carcasses were eviscerated, immersion chilled at 2 degrees C for 30 min, individually bagged, and stored for 1 wk at 2 degrees C, after which all carcasses were rinsed again, and spoilage bacteria in the rinsate were enumerated. There were no significant differences (P < or = 0.05) between the featherless and feathered broilers in numbers of aerobic bacteria, E. coli, and Campylobacter in rinse samples taken immediately after defeathering and no differences between carcasses picked for 30 or 60 s. There were no differences in numbers of spoilage bacteria after 1 wk of refrigeration for any of the feather presence-picking length combinations. Although the defeathering step in poultry processing has been identified as an opportunity for bacterial contamination from the intestinal tract and cross-contamination between carcasses, the presence of feathers and feather follicles does not make a significant difference in carcass bacterial contamination immediately after defeathering or in spoilage bacteria after 1 wk of refrigeration. PMID- 15339025 TI - Thickness and compositional effects on surface heating rate of bologna during in package pasteurization. AB - The surface heating rate (gamma) and final surface temperature (alpha) during in package pasteurization were determined for different thickness levels of 2 types of bologna having different (13 and 18%) fat contents. Three thicknesses (4, 12, and 20 mm), corresponding to 1, 3, and 5 slices of bologna, were vacuum-packaged separately in a clear polymer pouch after placing a thermocouple on the surface. Refrigerated samples were immersed in a water bath set to 1 of 4 predetermined temperatures (60, 70, 80, and 90 degrees C), and time and temperature data were recorded for 10 min. Surface heating rate was fastest in the thinnest (4 mm) and slowest in the thickest (20 mm) samples for all 4 temperatures. Surface heating rate was slower in bologna with the higher fat content compared with the lower fat bologna. Final surface temperature attained after 3 min was lower with increased thickness levels for all temperatures. Thus, meat sample thickness and fat content significantly affect surface heating rate and final surface temperature during in-package pasteurization of bologna. PMID- 15339026 TI - Influence of irradiation and storage on the quality of ready-to-eat turkey breast rolls. AB - Influence of irradiation and storage on the quality of ready-to-eat (RTE) turkey breast rolls was investigated. Commercial oven roasted turkey breast rolls purchased from local stores were sliced and vacuum packaged. The sliced samples were randomly divided into 3 groups and irradiated at 0, 1.0, or 2.0 kGy using a linear accelerator. Color, 2-TBA-reactive substances (TBARS), sensory characteristics, and volatiles were evaluated at 0, 7, and 14 d of storage. Irradiation increased color a* value of turkey breast rolls. Irradiation and storage did not influence TBARS values. Sensory evaluation showed that irradiation significantly increased sulfury flavor. Because a dramatic increase in sulfur compounds was detected in irradiated samples, the sulfury flavor should be due to the sulfur compounds formed during irradiation. Irradiation also increased the amounts of acetylaldehyde, 2-methyl butanal, 3-methyl butanal, benzene, and toluene. It was concluded that irradiation significantly influenced the odor and flavor of RTE turkey breast rolls under vacuum packaging conditions. Therefore, strategies to prevent negative changes in the quality of irradiated RTE turkey breast rolls are needed. PMID- 15339027 TI - Prediction of physical, color, and sensory characteristics of broiler breasts by visible/near infrared reflectance spectroscopy. AB - The feasibility of predicting pH, color, shear force, and sensory characteristics of chicken breasts deboned at 2, 4, 6, and 24 h postmortem by visible/near infrared reflectance spectroscopy (NIRS) in the 400 to 1850 nm region was determined. Prediction of physical attributes of Commission Internationale de l'Eclairage (CIE) color values (L*, a*, and b*), pH, and shear force had better accuracies than those of individual sensory attributes. Calibration and validation statistics for shear force and sensory traits indicated that visible/near infrared models were not significantly improved for cooked muscles compared with predictions based on raw muscle characteristics. On the basis of predicted shear values from the partial least squares (PLS) model, breast samples were classified into "tender" and "tough" classes with a correct classification of 74.0% if the boundary was set at 7.5 kg. The model developed from measured shears using soft independent modeling of class analogy/principal components analysis (SIMCA/PCA) showed nearly the same classification success. PMID- 15339028 TI - Inhibitors of topoisomerases as anticancer drugs: problems and prospects. AB - DNA topoisomerases, which solve topological problems associated with various DNA transactions, are the targets of many therapeutic agents. Various topoisomerase inhibitors especially, topo-poisons, camptothecin (topo-I) and etoposide (topo II) are some of the drugs that are used in the current treatment protocols, particularly for the treatment of leukemia (AML, ALL etc). However, tumor resistance, normal and non-specific tissue cytotoxicity are the limitations for successful development of these drugs as one of the primary therapeutic agents for the treatment of tumors in vitro. This brief review presents the current understanding about cytotoxicity development and outlines various approaches to overcome the limitations for enhancing the efficacy of topo-poison based anticancer drugs. PMID- 15339029 TI - Anti-HIV and cytotoxic ruthenium(II) complexes containing flavones: biochemical evaluation in mice. AB - Ru(II) polypyridyl complexes containing 3-hydroxyflavone derivatives as coligands were screened for anti-HIV and cytotoxic activities against eleven tumor cell lines. In order to check the effect of flavones containing Ru(II) complexes in vivo on a mammal, a representative complex Ru(L)2(DMSO)2 x 5H2O (LH-3-Hdroxy-4' benzyloxyflavone; M5) was orally administered to adult male mice. Its effects on protein content and LDH were studied in different tissues of the animal. The compound got absorbed and retained in the blood between 1-3 hr after feeding. As compared to the normal and DMSO control sets, tissue specific significant reversible changes in the protein content as well as in LDH activity were observed between 1-4 hr of treatment. However, on polyacrylamide gel electrophoresis, except some tissue specific transitory alterations, expression patterns of five LDH isozymes were unchanged after feeding the compound. The present results suggested that in addition to its potent cytotoxic and anti-HIV effects on cell lines in vitro, M5 inhibited LDH activity, but reversibly with a little effect on biosynthetic status of the enzyme in mice. PMID- 15339030 TI - Effect of 5-lipoxygenase inhibition on events associated with inflammatory bowel disease in rats. AB - Leukotrienes play a part in inflammatory response. The unique role of the enzyme 5-lipoxygenase (5-LOX) in the production of leukotrienes makes it a likely therapeutic target for inflammatory conditions like asthma, rheumatoid arthritis, psoriasis, and inflammatory bowel disease (IBD). The aim of the present study was to evaluate the effect of zileuton, an orally active selective 5-LOX inhibitor against the events associated with dextran sodium sulphate-induced colitis in a rat model of IBD. The animals were administered simultaneously zileuton (100mg/kg) or sulphasalazine (100mg/kg) orally for 7 days. On day eight, rats were sacrificed, and distal colon isolated to determine myeloperoxidase activity, in vivo superoxide dismutase activity, prostaglandin E2 levels and histological examination. Both zileuton and sulphasalazine significantly prevented the development of inflammatory events associated with colitis. The effect of zileuton was more pronounced towards reducing myeloperoxidase activity and increasing PGE2 levels in distal colon. The results show that chemotactic leukotrienes are responsible for inflammatory surge in damaged colon and, zileuton, significantly improved healing by inhibition of neutrophil recruitment and indirectly through increase in prostaglandins at the site of inflammation. It is suggested that inhibitors of 5-LOX enzyme may have useful therapeutic role in the treatment of chronic intestinal inflammation. PMID- 15339031 TI - Anti-ulcer and antioxidant activity of Normacid, a herbomineral formulation. AB - Effect of various doses (125, 250, 500 and 1000 mg/kg, po) of Normacid was studied on gastric secretion and gastric ulcers in pylorus-ligation and on ethanol-induced gastric mucosal injury in rats. The reduction in ulcer index in both the models along with the reduction in total acidity and an increase in the pH of gastric fluid in pylorus-ligated rats proved the anti-ulcer activity of Normacid. The increase in the levels of superoxide dismutase, catalase, reduced glutathione and membrane bound enzymes like Ca2+ ATPase, Mg2+ ATPase and Na+K+ ATPase and decrease in lipid peroxidation in both the models showed the antioxidant activity of the formulation. Thus it can be concluded that the anti ulcer activity shown by Normacid may be due to the modulation of defensive factors by improvenent in gastric cytoprotection and partly due to antioxidant property. PMID- 15339032 TI - Effect of trace elements on surface hydrophobicity and adherence of Escherichia coli to uroepithelial cells. AB - Trace elements have significant effect on the physiology of bacteria. Variation in the concentration of trace elements may affect the expression of virulence by microorganisms. The effect of trace elements on hydrophobicity and adherence of E.coli to uroepithelial cells was studied. Increasing concentrations of Ca2+, Mg2+, Fe3+ and Zn2+ significantly decreased the surface hydrophobicity. Toxic trace elements like Co2+, Cu2+, Mn2+ and Ni2+ did not alter surface hydrophobicity. With regards to adherence of E.coli to uroepithelial cells, only Mg2+ had significant effect. Toxic trace elements decreased the rate of cell adherence. The pathogenic strains of E.coli showed higher surface hydrophobicity and better cell adherence compared to the nonpathogenic strains. There was good correlation between surface hydrophobicity and cell adherence at higher concentrations (0.1 to 0.2mM) of Fe2+ and Zn2+. The results indicated that trace elements can significantly affect surface hydrophobicity and adherence of E.coli to uroepithelial cells. Such effect may have a significant impact on the initial stages of bacterial infection. PMID- 15339033 TI - Effect of lupeol isolated from Crataeva nurvala Buch.-Ham. stem bark extract against free radical induced nephrotoxicity in rats. AB - Lupeol, isolated from Crataeva nurvala stem bark in doses 40 and 80 mg/kg body weight, po, for 10 days, decreased the concentration of blood urea nitrogen, creatinine and lipid peroxidation and increased glutathione and catalase activities in cisplatin (5 mg/kg body weight, ip) induced nephrotoxicity in rats. The increased glutathione and catalase activities are indicative of antioxidant properties of lupeol. PMID- 15339034 TI - Effect of dietary n-3 and n-6 fatty acids on tobramycin induced nephrotoxicity. AB - Post fish oil(n-3 fatty acids) treatment (5mg/kg/day for 12 days) was effective in bringing the reversal of tobramycin (160mg/kg/day,ip for 12 days) induced nephrotoxicity in albino rats as was evident by normal urea, creatinine, cholesterol and inorganic phosphate levels in the serum of the treatment group compared with group receiving tobramycin only. The return of normal levels of alkaline and acid phosphatase in kidney homogenates of post fish oil treatment group also indicated the beneficial effect of dietary n-3 fatty acids(fish oil) more than n-6 fatty acids(olive oil).The results suggest that oral supplements of dietary n-3 fatty acids (fish oil) for nearly two weeks after tobramycin exposure is more beneficial than n-6 fatty acids (olive oil) as it results in reversal of nephrotoxicity induced by tobramycin. PMID- 15339035 TI - Evaluation of protective potentials of a potentized homeopathic drug, Chelidonium majus, during azo dye induced hepatocarcinogenesis in mice. AB - Several cytogenetical and enzymatic protocols were used to test if two microdoses of Chelidonium majus, namely Chelidonium-30 (Ch-30) and Chelidonium-200 (Ch-200), used as homeopathic drugs, showed anti-tumor activity and also favorably modulated genotoxic damages produced by an azo dye in mice at several intervals of fixation. Different sets of healthy mice were fed: (i) hepatocarcinogen, p dimethylaminoazobenzene (p-DAB, initiator) + phenobarbital (PB, promoter), (ii) only p-DAB, (iii) only PB, and (iv) neither p-DAB nor PB (normal control). Mice fed with p-DAB + PB were divided into different sets that were also fed either Ch 30 (v) or Ch-200 (vi) or diluted alcohol (vii), the "vehicle" of the microdoses of Chelidonium. All mice of group (i), a few of group (ii) and group (vii) and none of groups (iii) and (iv) developed tumors in liver at the longer intervals of fixation. The frequencies of chromosome aberrations (CA), micronucleated erythrocytes (MN), mitotic index (MI) and sperm head abnormality (SHA) were much higher in groups (i) and (vii) mice than in groups (ii), (iii) and (iv) mice at all fixation intervals. However, in mice of both groups (v) and (vi), the frequencies of CA, MN, SHA were strikingly less than those of groups (i) and (vii), and moderately less than those of groups (ii) and (iii). Both Ch-30 and Ch 200 also modulated favourably some toxicity marker enzymes like acid and alkaline phosphatases, peroxidases, glutamate oxaloacetate and glutamate pyruvate transaminases in liver, kidney and spleen tissues of the carcinogen fed mice. The microdoses of Chelidonium having no visible ill effects of their own, may be strong candidates for use in delaying/protecting liver cancer. PMID- 15339036 TI - Isolation and identification of marine chitinolytic bacteria and their potential in antifungal biocontrol. AB - Chitinolytic marine bacterial strains (30) were isolated from the sea dumps at Bhavnagar, India. They were screened as chitinase producers on the basis of zone of clearance on chitin agar plates incorporated with calcofluor white M2R for the better resolution. Out of these, three strains namely, Pseudomonas sp., Pantoea dispersa and Enterobacter amnigenus showed high chitinase production. They were also found to produce proteases and therefore have a good potential for use as antifungal biocontrol agents for the control of fungal plant pathogens. These strains could degrade and utilize the mycelia of Macrophomina phaseoliena (Tassi) Goidanich and Fusarium sp. In vitro, these strains could inhibit the growth of Fusarium sp. and M. phaseolina. The culture filtrate inhibiting hyphal elongation was observed microscopically. PMID- 15339037 TI - Biochemical changes in Glomus fasciculatum colonized roots of Lycopersicon esculentum in presence of Meloidogyne incognita. AB - Glasshouse experiments were conducted to elicit biochemical substantiation for the observed difference in resistance to nematode infection in roots colonized by mycorrhiza, and susceptibility of the fresh flush of roots of the same plant that escaped mycorrhizal colonization. Tomato roots were assayed for their biochemical profiles with respect to total proteins, total phenols, indole acetic acid, activities of polyphenol oxidase, phenylalanine ammonia lyase and indole acetic acid oxidase. The roots of the same plant (one set) received Glomus fasciculatum and G. fasciculatum plus juveniles of Meloidogyne incognita separately; and half the roots of second set of plants received G. fasciculatum while the other half of roots did not receive any treatment. Roots colonized by G. fasciculatum recorded maximum contents of proteins and phenols followed by that of the roots that received G. fasciculatum plus M. incognita. However, IAA content was lowest in the roots that received mycorrhiza or mycorrhiza plus juveniles of root-knot nematode and correspondingly. Roots that received juveniles of root-knot nematode recorded maximum IAA content and per cent increase over healthy check and mycorrhiza-inoculated roots. The comparative assay on the activities of PPO, PAL and IAA oxidase enzymes in treated and healthy roots of tomato, indicated that PAL and IAA oxidase activities were maximum in G. fasciculatum colonized roots followed by the roots that received mycorrhiza plus juveniles of root-knot nematode, while the activity of PPO was minimum in these roots. The roots that received juveniles of root-knot nematode recorded minimum PAL and IAA oxidase activities and maximum PPO activity. Since the roots of same plant that received mycorrhiza and that did not receive mycorrhiza; and the plant that received nematode alone and mycorrhiza plus nematode recorded differential biochemical contents of proteins, total phenols and IAA, and differential activities of enzymes under study, it was evident that the biochemical defense response to mycorrhizal colonization against root-knot nematodes was localized and not systemic. This explained for the response of plant that differed in root galling due to nematode infection in presence of mycorrhizal colonization. The new or fresh roots which missed mycorrhizal colonization, got infected by nematodes and developed root galls. PMID- 15339038 TI - Plant defense induced in in vitro propagated banana (Musa paradisiaca) plantlets by Fusarium derived elicitors. AB - Perception of microbial signal molecules is part of the strategy evolved by plants to survive attacks by potential pathogens. To gain a more complete understanding of the early signaling events involved in these responses, we used fungal components of Fusarium under in vitro condition and checked the rise in signal molecule, salicylic acid (SA), and marker enzymes in defense reactions against the pathogen. SA level increased by 21 folds in elicitor treated plantlets as compared to that of control plantlets and there was marked increase in phenylalanine ammonia-lyase(PAL), peroxidase(POX), polyphenol oxidase(PPO) along with higher total phenolic content. Present results indicated that use of fungal components had successfully induced systemic resistance in in vitro cultured banana plantlets. PMID- 15339039 TI - Toxicological studies of pesticides on cytoplasmic streaming in Nitella. AB - In the present study, changes in velocity of cytoplasmic streaming in the giant internodal cells of Nitella for varying concentration of the pesticides, 2,4-D, dieldrin, malathion, methyl parathion and endosulfan, were measured. Marked decrease in the velocity of cytoplasmic streaming was found at the concentrations of 0.01, 0.1, 1, 10 and 100mM. Dieldrin was the most toxic to all the pesticides investigated, followed by methyl parathion, endosulfan, malathion and 2,4-D. Threshold values for dieldrin, methylparathion, endosulfan, malathion and 2,4-D as indicated by the onset of decrease in the normal cytoplasmic streaming velocity were less than 6.25 x 10(-6), 2.5 x 10(-5), 5 x 10(-5), 5 x 10(-5) and 1.25 x 10(-5)M respectively. Cessation of streaming was noticed above 1mM in dieldrin and above 10mM when exposed to methylparathion and endosulfan. Cessation of streaming was not seen up to 100mM concentration of 2,4-D and malathion. PMID- 15339040 TI - Protective effect of ginger, Zingiber officinale Rosc on experimental atherosclerosis in rabbits. AB - The effects of air dried ginger powder (0.1g/kg body weight, po, for 75 days) were studied on experimentally induced atherosclerosis in rabbits by cholesterol feeding (0.3g/kg body weight, po). Cholesterol feeding for 75 days lead to distinct development of atheroma in the aorta and coronary arteriesof the rabbits and this was significantly inhibited by about 50% following ginger administration. There was distinct decrease in lipid peroxidation and enhancement of fibrinolytic activity in ginger treated animals. However, ginger did not lower blood lipidsto any significant extent. This distinct protection from the development of atherosclerosis by ginger is probably because of its free radical scavanging, prostaglandin inhibitory and fibri properties. PMID- 15339041 TI - Immediate placement and loading of implants in extraction sites: procedures in the aesthetic zone. PMID- 15339043 TI - CXCR4-SDF-1 signalling, locomotion, chemotaxis and adhesion. AB - Chemokines, small pro-inflammatory chemoattractant cytokines, that bind to specific G-protein-coupled seven-span transmembrane receptors present on plasma membranes of target cells are the major regulators of cell trafficking. In addition some chemokines have been reported to modulate cell survival and growth. Moreover, compelling evidence is accumulating that cancer cells may employ several mechanisms involving chemokine-chemokine receptor axes during their metastasis that also regulate the trafficking of normal cells. Of all the chemokines, stromal-derived factor-1 (SDF-1), an alpha-chemokine that binds to G protein-coupled CXCR4, plays an important and unique role in the regulation of stem/progenitor cell trafficking. First, SDF-1 regulates the trafficking of CXCR4+ haemato/lymphopoietic cells, their homing/retention in major haemato/lymphopoietic organs and accumulation of CXCR4+ immune cells in tissues affected by inflammation. Second, CXCR4 plays an essential role in the trafficking of other tissue/organ specific stem/progenitor cells expressing CXCR4 on their surface, e.g., during embryo/organogenesis and tissue/organ regeneration. Third, since CXCR4 is expressed on several tumour cells, these CXCR4 positive tumour cells may metastasize to the organs that secrete/express SDF-1 (e.g., bones, lymph nodes, lung and liver). SDF-1 exerts pleiotropic effects regulating processes essential to tumour metastasis such as locomotion of malignant cells, their chemoattraction and adhesion, as well as plays an important role in tumour vascularization. This implies that new therapeutic strategies aimed at blocking the SDF-1-CXCR4 axis could have important applications in the clinic by modulating the trafficking of haemato/lymphopoietic cells and inhibiting the metastatic behaviour of tumour cells as well. In this review, we focus on a role of the SDF-1-CXCR4 axis in regulating the metastatic behaviour of tumour cells and discuss the molecular mechanisms that are essential to this process. PMID- 15339044 TI - IGF-I receptor, cell-cell adhesion, tumour development and progression. AB - The insulin-like growth factor I receptor (IGF-IR) has been implicated in the development and progression of many common cancers and other neoplastic diseases. The tumorigenic potential of IGF-IR relies on its antiapoptotic and transforming activities. The molecular mechanisms by which IGF-IR controls the proliferation and survival of tumour cells have been extensively studied and many pathways have been delineated. However, the role of IGF-IR in the regulation of non-mitogenic cell functions is less well understood. Here we focus on IGF-IR-dependent cell cell adhesion. Limited studies suggested that IGF-IR can regulate cell aggregation and intercellular adhesion mediated by cadherins and cadherin associated proteins. We review the mechanisms of this process and discuss the impact of IGF-IR-dependent cell-cell adhesion on the phenotype of tumour cells. PMID- 15339042 TI - CD44 in cancer progression: adhesion, migration and growth regulation. AB - It is well established that the large array of functions that a tumour cell has to fulfil to settle as a metastasis in a distant organ requires cooperative activities between the tumour and the surrounding tissue and that several classes of molecules are involved, such as cell-cell and cell-matrix adhesion molecules and matrix degrading enzymes, to name only a few. Furthermore, metastasis formation requires concerted activities between tumour cells and surrounding cells as well as matrix elements and possibly concerted activities between individual molecules of the tumour cell itself. Adhesion molecules have originally been thought to be essential for the formation of multicellular organisms and to tether cells to the extracellular matrix or to neighbouring cells. CD44 transmembrane glycoproteins belong to the families of adhesion molecules and have originally been described to mediate lymphocyte homing to peripheral lymphoid tissues. It was soon recognized that the molecules, under selective conditions, may suffice to initiate metastatic spread of tumour cells. The question remained as to how a single adhesion molecule can fulfil that task. This review outlines that adhesion is by no means a passive task. Rather, ligand binding, as exemplified for CD44 and other similar adhesion molecules, initiates a cascade of events that can be started by adherence to the extracellular matrix. This leads to activation of the molecule itself, binding to additional ligands, such as growth factors and matrix degrading enzymes, complex formation with additional transmembrane molecules and association with cytoskeletal elements and signal transducing molecules. Thus, through the interplay of CD44 with its ligands and associating molecules CD44 modulates adhesiveness, motility, matrix degradation, proliferation and cell survival, features that together may well allow a tumour cell to proceed through all steps of the metastatic cascade. PMID- 15339045 TI - Tumour biological aspects of CD24, a mucin-like adhesion molecule. AB - CD24 is a molecule that recently has raised considerable attention in tumour biology. It is involved in cell adhesion and metastatic tumour spread. It has also been described as a new diagnostic marker of tumours, of neuroendocrine differentiation and, possibly most intriguing of all, of patient prognosis. High rates of CD24 expression detected by immunohistochemistry have been found in epithelial ovarian cancer (83%), breast cancer (85%), non-small cell lung cancer (45%), prostate cancer (48%) and pancreatic cancer (72%). With the exception of pancreatic cancer, high rates of CD24 are significantly associated with a more aggressive course of the disease, a finding that remains significant in a multivariate analysis. The aim of this review is to summarize relevant work covering these aspects of CD24. PMID- 15339046 TI - Role of subtilisin-like convertases in cadherin processing or the conundrum to stall cadherin function by convertase inhibitors in cancer therapy. AB - Cadherins are a family of intercellular adhesion receptors. Produced as inactive precursors, they become functional adhesion molecules after proteolytic cleavage by subtilisin-like pro-protein convertases (PCs). Owing to their activation and assembly into multiprotein adhesion complexes at sites of cell contacts, adhesion competent cadherins are prerequisite for tissue integrity. In recent years evidence has accumulated that intercellular junctions not only provide mechanical linkage, but in addition are potent modulators of signalling cascades. This infers a biological role to intercellular adhesion complexes that is significantly more complex and powerful. Currently, the broad implications of disturbances in somatic tissue adhesion components are only just beginning to emerge. Prominent examples of adhesion defects include autoimmune diseases, or tumour invasion and metastasis and malignant transformation. This review reports on our current knowledge of cadherin function and their maturation by pro-protein convertases, and puts special emphasis on the consequences of pro-protein convertase inhibition for epithelial tissue homeostasis. PMID- 15339047 TI - Laminin-5 in epithelial tumour invasion. AB - Laminin-5 (LN-5), consisting of alpha3-, beta3-, and gamma2-chains, is a component of the cell adhesion complex containing hemidesmosomes and anchoring fibrils. This protein is a major constituent of the extracellular matrix and has recently proved to be an invasion marker for epithelial cells in many immunohistochemical surveys, indicating that it is frequently expressed in the invading edges of epithelial tumour cells. Additionally, intracellular accumulation of monomeric gamma2-chains has been widely observed in the invasive carcinoma cells, but its mechanism was not entirely understood. Epithelial carcinoma cells prefer to adhere onto the LN-5-rich basement membranes using the specific integrins as receptors. Induction of cell migration is an important function of LN-5 and the enhanced activity is observed in its truncated form after proteolytic shedding of the N-terminal fragments of gamma2-chains. This processing was demonstrated to be mediated mainly by several kinds of matrix metalloproteinases. The degraded fragments of gamma2-chains, released from invading carcinomas, can be immunodetected in biological fluids and potentially utilized in the clinical diagnosis of various epithelial cancers. Here, we summarize the previous clinical investigations of LN-5 in epithelial tumour progression, and also discuss what it can regulate in the cell physiological events. PMID- 15339048 TI - Cell-cell adhesion molecule CEACAM1 is expressed in normal breast and milk and associates with beta1 integrin in a 3D model of morphogenesis. AB - CEA cell adhesion molecule-1 (CEACAM1) is a cell-cell adhesion molecule that, paradoxically, is expressed in an apical location in normal breast epithelium. Strong lumenal membrane staining is observed in 100% of normal glands (11/11), low in atypical hyperplasia (2/6), high in cribiform ductal carcinoma in situ (DCIS) (8/8), but low in other types of DCIS (2/15). Although most invasive ductal carcinomas express CEACAM1 (21/26), the staining pattern tends to be weak and cytoplasmic in tumours with minimal lumena formation (grades 2-3), while there is membrane staining in well-differentiated tumours (grade 1). The 'normal' breast epithelial line MCF10F forms acini with lumena in Matrigel with apical membrane expression of CEACAM1. MCF7 cells that do not express CEACAM1 and fail to form lumena in Matrigel, revert to a lumen forming phenotype when transfected with the CEACAM1-4S but not the -4L isoform. CEACAM1 directly associates with and down-regulates the expression of beta1-integrin. Immuno-electron microscopy reveals numerous vesicles coated with CEACAM1 within the lumena, and as predicted by this finding, CEACAM1 is found in the lipid fraction of breast milk. Thus, CEACAM1 is a critical molecule in mammary morphogenesis and may play a role in the absorption of the lipid vesicles of milk in the infant intestinal tract. PMID- 15339049 TI - Curcumin, a multi-functional chemopreventive agent, blocks growth of colon cancer cells by targeting beta-catenin-mediated transactivation and cell-cell adhesion pathways. AB - Colorectal cancer, the second most frequent diagnosed cancer in the US, causes significant morbidity and mortality in humans. Over the past several years, the molecular and biochemical pathways that influence the development of colon cancer have been extensively characterized. Since the development of colon cancer involves multi-step events, the available drug therapies for colorectal cancer are largely ineffective. The radiotherapy, photodynamic therapy, and chemotherapy are associated with severe side effects and offer no firm expectation for a cure. Thus, there is a constant need for the investigation of other potentially useful options. One of the widely sought approaches is cancer chemoprevention that uses natural agents to reverse or inhibit the malignant transformation of colon cancer cells and to prevent invasion and metastasis. Curcumin (diferuloylmethane), a natural plant product, possesses such chemopreventive activity that targets multiple signalling pathways in the prevention of colon cancer development. PMID- 15339050 TI - The role of altered cell-cell communication in melanoma progression. AB - Under normal homeostasis, melanocyte growth and behaviour is tightly controlled by the surrounding keratinocytes. Keratinocytes regulate melanocyte behaviour through a complex system of paracrine growth factors and cell-cell adhesion molecules. Pathological changes, leading to development of malignant melanoma, upset this delicate homeostatic balance and can lead to altered expression of cell-cell adhesion and cell-cell communication molecules. In particular, there is a switch from the E-cadherin-mediated keratinocyte-melanocyte partnership to the N-cadherin-mediated melanoma-melanoma and melanoma-fibroblast interaction. Other changes include the alteration in the gap junctions formed between the melanocyte and keratinocyte. Changes in the connexin expression, in particular the loss of connexin 43, may result in a reduction or a loss of gap junctional activity, which is thought to contribute towards tumour progression. In the current review we describe the alterations in cell-cell adhesion and communication associated with melanoma development and progression, and discuss how a greater understanding of these processes may aid the future therapy of this disease. PMID- 15339051 TI - New insights into syndecan-2 expression and tumourigenic activity in colon carcinoma cells. AB - Adhesion receptors play crucial roles in the neoplastic transformation of normal cells through induction of cancer-specific cellular behaviour and morphology. This implies that cancer cells likely express and utilize a distinct set of adhesion receptors during carcinogenesis. Colon cancer is an excellent model system for the study of this process, since both molecular genetic and morphological changes have been well established for this disease. We recently reported increased expression of the cell surface adhesion receptor, syndecan-2, in several colon carcinoma cell lines. Indeed, increased syndecan-2 expression was necessary for tumourigenic activity, suggesting that syndecan-2 might have value as both a new diagnostic marker and a possible therapeutic target. Here, we review recent advances in understanding the role of syndecan-2 in the carcinogenesis of colon cells, and discuss a leading role for this molecule in a new era for colon cancer treatment. PMID- 15339053 TI - New investigative regimens and cytotoxic agents in thoracic cancers: gemcitabine and pemetrexed. AB - Several new antimetabolites, administered alone or in combination, are changing the therapeutic landscape for thoracic cancer. Two-drug combinations involving these newer drugs are becoming the standard of care for non-small-cell lung cancer (NSCLC), largely due to improvements in survival rates, time to disease progression, and response rates as well as an improved safety profile. Gemcitabine (Gemzar) has elicited considerable interest in this disease, as a combination partner in chemotherapeutic regimens. Another promising agent is pemetrexed (Alimta), a folate-based inhibitor of thymidylate synthase. In preclinical development, pemetrexed both alone and in combination with other cytotoxic agents has exhibited activity across a broad range of tumor models, including NSCLC and mesothelioma. In clinical trials of patients with NSCLC, pemetrexed has been an effective, well-tolerated agent that can be used as monotherapy or in combination with other agents at full dose. In clinical trials of patients with mesothelioma, the combination of pemetrexed and cisplatin demonstrated a significant improvement in survival, response, and patient quality of-life parameters. The principle toxicities of pemetrexed can be minimized by folate and vitamin B12 supplements. PMID- 15339054 TI - Current paradigms in first-line treatment of non-small-cell lung cancer. AB - Standard first-line chemotherapy for the majority of patients with advanced non small-cell lung cancer (NSCLC) consists of platinum-based combination regimens including one of the newer-generation agents, such as gemcitabine (Gemzar), a taxane, vinorelbine (Navelbine), or irinotecan (Camptosar). Several effective regimens are available, the choice of which will depend on treatment goals, individual patient or disease factors, as well as physician preferences. This paper surveys randomized trials of many of the newer-generation chemotherapy combinations in patients with advanced NSCLC to examine several issues, such as which new-generation regimen to use, whether a platinum agent is needed, the optimal number of drugs in the combination, and treatment duration. PMID- 15339052 TI - Integrin signalling and the cellular response to ionizing radiation. AB - Cell survival and cycling in mammalian cells are both greatly affected by ionizing radiation and are both strictly controlled by integrated integrin mediated adhesion to extracellular matrix (ECM) proteins and by binding of growth factors to their cognate receptors. Recent emerging findings show a diverse panel of integrin-dependent signals that are channelled into the regulation and modification of the cellular response to ionizing radiation. Cell adhesion mediated radioresistance and alteration of DNA damage-induced cell cycle arrest in cells attached to the ECM can be linked to focal adhesion protein signalling. This review summarizes the latest radiobiological and radiooncological findings about integrins and their signal transduction pathways. PMID- 15339055 TI - Carboplatin/gemcitabine combination in advanced NSCLC. AB - The treatment of advanced non-small-cell lung cancer (NSCLC) has evolved rapidly over the past few years. Systemic chemotherapy is associated with both quality of life and modest survival benefit for patients with advanced NSCLC. Platinum-based doublet combinations are the "standard of care." The US Food and Drug Administration (FDA) has approved gemcitabine (Gemzar), a pyrimidine analog, to be used in combination with cisplatin for the treatment of advanced NSCLC in the first-line setting. Randomized clinical trials have established comparable efficacy with improved therapeutic index for the carboplatin/gemcitabine regimen when compared with cisplatin/gemcitabine and other platinum doublets. Nonhematologic toxicities occur at a lower frequency with carboplatin/gemcitabine combinations compared with other "standard" platinum-based doublets, whereas dose limiting thrombocytopenia, the most common toxicity, rarely requires therapeutic intervention. Both the 3- and 4-week schedules of carboplatin/gemcitabine result in similar efficacy and toxicity profiles, but the 3-week regimen is preferred. The combination of carboplatin and gemcitabine is an effective regimen with an acceptable toxicity profile for the treatment of advanced NSCLC. This regimen can also be used as a foundation for the development of innovative combinations with molecularly targeted agents. PMID- 15339056 TI - Gemcitabine-containing regimens vs others in first-line treatment of NSCLC. AB - Standard first-line chemotherapy regimens in advanced non-small-cell lung cancer (NSCLC) include carboplatin (Paraplatin)/paclitaxel, cisplatin/docetaxel (Taxotere), cisplatin/gemcitabine (Gemzar), and cisplatin/vinorelbine (Navelbine). An informal meta-analysis of 13 randomized trials of these regimens in NSCLC indicates no marked differences in terms of response rates or survival, but toxicity advantages with cisplatin/gemcitabine and cisplatin/vinorelbine regimens. An informal meta-analysis to assess the feasibility of substituting carboplatin for cisplatin in combination with gemcitabine or docetaxel shows no marked differences in efficacy between cisplatin- and carboplatin-containing regimens, although a slight trend favoring carboplatin/gemcitabine treatment may be observed; comparison of toxicity profiles among carboplatin-based regimens suggests advantages for carboplatin/gemcitabine treatment. A formal meta-analysis of 13 trials comparing gemcitabine/platinum combinations with other platinum based regimens in NSCLC indicates significant improvements in progression-free survival and overall survival with gemcitabine/platinum treatment. On balance, available data suggest that carboplatin/gemcitabine may be the first-line option with the best therapeutic index. PMID- 15339057 TI - Induction therapy for early-stage non-small-cell lung cancer. AB - Data from adjuvant trials clearly indicate that one of the most important problems in patients with resected non-small-cell lung cancer (NSCLC) is compliance to chemotherapy. In the postoperative setting, significant comorbidities and incomplete recovery after surgery often make it difficult for patients to tolerate or comply with systemic therapy. Therefore, it may be preferable to deliver chemotherapy before surgery as "neoadjuvant" or "induction" chemotherapy. The rationale for using induction chemotherapy is based on evidence that chemotherapy might reduce tumor burden and possess activity against micrometastases, resulting in improved results by surgery, radiotherapy, or a combination. Moreover, induction therapy facilitates in vivo assessment of tumor response or resistance. Potential drawbacks include the risk of perioperative complications, and the possibility that the tumor mass may become unresectable due to disease progression. During the past decade, four phase III randomized trials evaluated the role of induction chemotherapy in stage IIIA NSCLC. The first three studies consistently showed that induction chemotherapy improves survival compared with surgery alone. More recently, a large phase III trial performed by French investigators suggested a survival benefit in stage I/II patients, but not stage IIIA. The high activity of new platinum-based chemotherapy--based on response rate and 1-year survival in advanced disease- reinforces the rationale for the use of these new combinations in early-stage NSCLC, especially for a subset of patients traditionally treated with surgery alone. Several phase III trials are currently evaluating the role of new doublets as induction chemotherapy; these are discussed in the article. The results of these ongoing phase III trials should help clarify the role of induction chemotherapy in early-stage disease. PMID- 15339058 TI - Chemoradiation in NSCLC: focus on the role of gemcitabine. AB - Research to identify the optimal drugs for use in chemoradiotherapy has led to the development of the potent radiosensitizing agent gemcitabine (Gemzar), which has exhibited excellent activity in non-small-cell cancer. When used in sequential chemoradiotherapy regimens, gemcitabine has been associated with response rates of 57% to 68%. A full dose of gemcitabine (1,000 mg/m2) can be safely used as induction therapy, and there is no definitive indication of enhancement of radiotoxicity. In addition, results from phase I/II trials support the efficacy of concurrent gemcitabine/radiation therapy in improving overall response rates and overall survival. Rates of 68%, 37%, and 28%, respectively, for 1-, 2-, and 3-year survival have been reported for gemcitabine/cisplatin chemotherapy administered concurrently with radiotherapy. Although the optimal dose has yet to be determined, a weekly dose of 300 mg/m2 appears to be effective with an acceptable toxicity level. Additional clinical trials are warranted to assess the long-term efficacy and safety of gemcitabine in combination with other chemotherapeutic agents and radiation therapy for treatment of non-small-cell lung cancer. PMID- 15339059 TI - Pemetrexed: its promise in treating non-small-cell lung cancer. AB - The use of chemotherapy in the treatment of early and advanced non-small-cell lung cancer (NSCLC) has increased during the past decade. One of the main reasons for the increased acceptance of chemotherapy is the development of several new cytotoxic agents with a unique mechanism(s) of action and high single-agent activity, combined with a favorable toxicity profile. Pemetrexed (Alimta) is a novel antifolate that inhibits several enzymes involved in DNA synthesis (thymidylate synthase [TS], dihydrofolate reductase [DHFR], and glycinamide ribonucleotideformyltransferase [GARFT]). Pemetrexed's toxicity is markedly reduced by folic acid and vitamin B12 supplementation. The compound has been studied extensively in various tumor types, including NSCLC. In NSCLC, pemetrexed at 500 mg/m2, every 3 weeks, given i.v. over 10 minutes, has shown promising activity, and can safely be administrated with vitamin supplementation. After registration, single-agent pemetrexed will certainly add to the chemotherapeutic options available for pretreated patients and will most likely change significantly chemotherapy prescriptions in second-line chemotherapy. In first line chemotherapy, the role of platinum-based and -free combination doublet chemotherapy with pemetrexed still needs to be defined. Phase II data indicate high efficacy combined with favorable toxicity for pemetrexed in combination with cisplatin, carboplatin (Paraplatin), oxaliplatin (Eloxatin), gemcitabine (Gemzar), and vinorelbine (Navelbine). This review summarizes the clinical experience obtained thus far during the early clinical development of pemetrexed in NSCLC. PMID- 15339060 TI - The emerging role of pemetrexed for the treatment of malignant mesothelioma. AB - Malignant mesothelioma is a devastating disease with an onset 20 to 60 years after exposure to asbestos. Although most cytotoxic agents have been evaluated for the treatment of mesothelioma, few single agents have consistently yielded response rates above 20%. Antimetabolites are the most active drugs against mesothelioma, and of these, the antifolate group is the most widely studied and effective. Pemetrexed (Alimta), a new antifolate, may be more active because of its different mechanism of action. Several clinical trials have evaluated pemetrexed alone and in combination with a platinum agent for patients with malignant mesothelioma. A pivotal phase III trial has demonstrated that combination chemotherapy with pemetrexed and cisplatin improves survival, response rate, pulmonary function, and quality of life compared with single-agent cisplatin. Additional trials are evaluating pemetrexed in the neoadjuvant setting and in combination with other cytotoxic and targeted agents. PMID- 15339061 TI - Pemetrexed in advanced NSCLC: a review of the clinical data. AB - The novel multitargeted antimetabolite pemetrexed (Alimta), recently approved by the US Food and Drug Administration for the treatment of mesothelioma when combined with cisplatin, is also active in first- and second-line non-small-cell lung cancer (NSCLC). In a phase III trial comparing single-agent pemetrexed vs docetaxel (Taxotere) as second-line therapy in advanced NSCLC, survival was shown to be comparable between these agents, but side effects were significantly less frequent and severe for patients who received pemetrexed. In the frontline setting, phase II studies have shown significant activity and a very favorable toxicity profile of the combination of pemetrexed with a platinum agent. Pemetrexed has been well tolerated at systemic doses as a radiosensitizer when given as concurrent chest radiation, and a phase I study is under way to assess its tolerability in combination with carboplatin (Paraplatin) in this setting. Pemetrexed is an important addition to the armamentarium of medicines used to treat thoracic malignancies, and merits study in combination with other drugs having novel mechanisms of action. PMID- 15339062 TI - Pemetrexed in previously treated non-small-cell lung cancer. AB - Several decades of chemotherapy trials in non-small-cell lung cancer (NSCLC) have clearly shown a survival benefit for chemotherapy over best supportive care. However, only short-lived responses are attained, with an average of four cycles of chemotherapy, before tumor progression is observed. Second-line chemotherapy has been demonstrated to improve outcome, with docetaxel (Taxotere) as the predominant cytotoxic drug. A recent randomized trial in second-line NSCLC indicated that the novel drug pemetrexed (Alimta) attained the same response, time to progression, and survival as docetaxel. This finding ushers in a new age in second-line treatment that can be further invigorated by the addition of targeted agents. Accumulated evidence indicates that overexpression of epidermal growth factor receptor and HER2/neu, which occurs frequently in NSCLC, leads to the deregulation of PI3K and MAPK, activating Akt and enhancing chemoresistance. Future clinical trials in NSCLC will include tailored and multitargeted therapy and pemetrexed represents a significant step forward in this direction. PMID- 15339063 TI - Going straight: the promise to end hidden waiting times. PMID- 15339064 TI - Long-term condition ideas. Power to the people. AB - We are on the brink of a revolution in patient involvement, but partnerships involving the NHS and support groups must keep pace with 'patient power' if people with chronic medical conditions are to get the best services PMID- 15339065 TI - Commissioning. Meet the new power in primary care. AB - Practices and PCTs will need to negotiate a variety of options when implementing practice-led commissioning. Available options include how much power is devolved to practices, what services they will be commissioning and the incentives on offer. Practices must be regulated by PCTs. Their decisions should also be made in line with the broad strategic aims of their PCTs. PMID- 15339066 TI - Finance. Noel Plumridge wonders whether PCTs need finance directors. PMID- 15339067 TI - HSJ people. Built to last. PMID- 15339068 TI - Towards optimal regimens for the UVB phototherapy of psoriasis: a mathematical model. AB - A mathematical model is described that predicts the response of psoriasis to a treatment course of UVB irradiation. The basis of the model is that UVB acts by a direct effect on keratinocytes and that cell cycle arrest is the major mode of action in the phototherapeutic response in psoriasis. Although it is unlikely that UVB causes resolution of psoriatic plaques through a single mechanism, the current model has been based on epidermal cell cycle arrest and entry into the terminal differentiation compartment because this is likely to be a significant rate-limiting factor in determining response to treatment. The model has been validated against results obtained from published clinical studies on narrowband (TL-01) UVB phototherapy for psoriasis. The principal outcomes of the model are that for a given erythemal response, the number of exposures required for clearance is almost independent of the frequency with which patients attend for treatment and that the higher the exposure dose per treatment, the more rapidly will clearance result. The model has been used to suggest optimal regimens for the treatment of outpatients and inpatients. PMID- 15339069 TI - Functional characterization of beta1-integrin-positive epidermal cell populations. AB - Epidermal keratinocytes are heterogeneous and can be divided into stem cells (strong beta1-integrin expression) with unlimited clonogenic potential, transient amplifying cells (weaker beta1-integrin expression) with restricted proliferative capacity and terminally differentiated cells (no beta1-integrin expression) that have lost the capacity to divide. We tested the hypothesis that cell kinetic characteristics of the epidermal subpopulations differ. Single cell suspensions from small human skin punch biopsies were sorted flow cytometrically into a beta1 integrin weakly positive (dim) and strongly positive (bright) subpopulation and the clonogenic potential was compared in cell culture experiments. Image analysis was used to determine growth characteristics of the colonies. We found that cell size in the beta1-integrin bright subpopulation increased when colonies aged, whereas this was constant in the dim subpopulation. The total number of colonies formed and the growth rate of the colonies were higher in the beta1-integrin dim cells than in the bright subpopulation. Experimental data from this study confirm the hypothesis that cell kinetic characteristics of beta1-integrin dim and bright cells are different. Combining flow cytometric sorting, cell culture and image analysis provides powerful means for phenotypical and functional characterization of epidermal subpopulations. PMID- 15339070 TI - Frequency and distribution pattern of melanocytic naevi in Swedish 8-9-year-old children. AB - The naevus profile was examined in a Swedish cohort of 8-9-year-old children; 524/545 individuals (96%) were examined (279 boys and 245 girls). There was a wide variation in the total number of naevi (0-79) and boys had more naevi than girls (median 9 and 7, respectively, p<0.01). No dysplastic naevi were found. Overall, 15/ 524 (3%) had at least one lesion clinically diagnosed as a congenital melanocytic naevus. Boys had more naevi on the face (median 1) and trunk (median 5) than girls (median 0 and 3, respectively, p<0.001). There was no difference in the number of naevi on the legs between the two sexes. The highest counts per unit surface area for both sexes were found on the back, chest and the lateral aspect of the arms, areas intermittently sun-exposed. Children with fair skin and light eye colours had significantly more naevi than those with darker colours but children with red hair had very few naevi. Children with one or more naevi on the buttocks (25%), dorsal surfaces of the feet (11%) or on the scalp (7%) had twice as many naevi in total compared with those without naevi in these regions. Children with naevi in all three regions (0.8%) had four times as many naevi in total. A relationship between total counts and counts on the back or lateral aspect of the arms was found (r2 = 0.59). Either of these two areas might be suitable for predicting total naevus counts. PMID- 15339071 TI - The vulvar skin microenvironment: influence of different panty liners on temperature, pH and microflora. AB - The aim of this study was to confirm findings that vapour-impermeable panty liners might impair skin climate, and to assess their impact on the skin microflora. Temperature, surface pH and aerobic microflora were measured on vulvar skin of 102 women. The mean skin temperature was 1.1 degrees C higher when using a vapour-impermeable panty liner compared with not using one. Use of panty liners with vapour-permeable back sheets and acidic cores resulted in skin temperature, pH and microflora levels that were very close to those observed in persons not using liners. The temperature, pH and total number of microorganisms were significantly lower for users of vapour-permeable panty liners than for users of vapour-impermeable ones (p <0.05, p<0.001 and p<0.001, respectively). The microorganism densities were usually higher when using the vapour-impermeable panty liner, but mean differences were minor. The use of panty liners seems not to imply a microbial risk for normal, healthy women. PMID- 15339072 TI - Seborrhoeic dermatitis in the elderly: inferences on the possible role of disability and loss of self-sufficiency. AB - The constant increase of life expectancy in countries with high standards of living raises growing concern about many aspects of skin morbidity in the elderly. As regards seborrhoeic dermatitis, literature yields prevalence estimates for elderly people that are consistently higher than those reported from the general population. This prompts speculations on whether factors inherently involved in senescence might have some role in the course of seborrhoeic dermatitis. In a series of 186 subjects, aged > or =65, strength of association of the skin disorder with age, sex, coexisting non-cutaneous conditions, and degree of loss of self-sufficiency in activities of daily living (ADL index) was evaluated by univariate and multivariate analysis. We found that age (OR: 1.14; p<0.001) and, more importantly, dependency in more than one ADL (OR: 30.2; p<0.0001) were independent explanatory variables of seborrhoeic dermatitis. These findings suggest that senescence per se might have some significance in the natural history of this type of eczema. PMID- 15339073 TI - Chronic idiopathic and chronic autoimmune urticaria: clinical and immunopathological features of 68 subjects. AB - Skin tests with autologous serum elicit an immediate wheal-and-flare response in about 30-50% of chronic idiopathic urticaria subjects, suggesting that an autoimmune mechanism might be involved in the pathogenesis of this disease. The aim of the present work, involving 68 subjects with chronic idiopathic urticaria, was to distinguish between the serum-positive and serum-negative cases and highlight the clinical differences between the two groups on the basis of the Breneman scale score. We also tried to correlate the finding of a positive response to the autologous serum skin test with other autoimmune diatheses or fully developed autoimmune disorders. Our results did not demonstrate any significant differences between the two groups with regard to mean age, sex distribution, angioedema and mucosal/cutaneous atopy. However, all subjects with positive autologous serum skin test presented more severe clinical features than serum-negative subjects. We found no differences between the two groups in the incidence of autoimmune disease. PMID- 15339074 TI - Are adverse skin reactions to cosmetics underestimated in the clinical assessment of contact dermatitis? A prospective study among 1075 patients attending Swedish patch test clinics. AB - It is known that cosmetics and skin care products can cause adverse skin reactions. However, the frequency of adverse reactions reported to the Medical Product Agency (MPA) in Sweden is low. The purpose of the present study was to evaluate the occurrence of adverse skin reactions to cosmetics among patients referred for standard patch testing owing to suspected contact dermatitis in general, most frequently hand eczema. Consecutive patients at four patch test clinics in Sweden were invited to participate; 1075 were included. Of these, 47.3% (54.2% women and 30.8% men) reported current or previous adverse skin reactions to cosmetics and skin care products. This group showed significantly more positive patch test reactions, a higher prevalence of atopic dermatitis and the dermatitis was more frequently located in the face and neck region. Our results show that patients referred for standard patch testing have--or have had- a large proportion of self-reported adverse reactions to cosmetics or skin care products. We conclude that among patients with suspected contact dermatitis, adverse reactions to cosmetics can be a more important aetiological and/or complicating factor than is commonly acknowledged and that the reporting of such reactions to the MPA probably can be improved. PMID- 15339075 TI - Clinical manifestations of cutaneous graft-versus-host disease after allogeneic haematopoietic cell transplantation: long-term follow-up results in a single Turkish centre. AB - The aim of this study was to evaluate the clinical manifestations of cutaneous graft-versus-host disease (GVHD) developed after allogeneic haematopoietic cell transplantation. In all, 67 patients were evaluated: 49 patients developed acute GVHD, 17 patients developed de novo chronic GVHD and 29 developed secondary chronic (15 limited, 14 progressive) GVHD following acute cutaneous GVHD. Of the 46 patients with chronic GVHD, lichenoid lesions were observed in 32 and sclerodermoid lesions were observed in 12. In four patients with sclerodermoid cutaneous GVHD, these lesions occurred after a lichenoid phase. Oral lesions were present in 61% of the patients and six of them had only oral mucosal involvement without any skin lesions. Nail lesions were observed in 31% of the patients. During the follow-up period 15 patients with GVHD died and in 7 of them the cause of death was related to chronic GVHD. In conclusion, GVHD has a wide spectrum of cutaneous manifestations, which can be used as an important tool for the early diagnosis of the disease. PMID- 15339076 TI - Treatment of chronic hand dermatoses with UVB-TL01. AB - UVB-TL01 is proven to be efficient in whole body treatment for several chronic inflammatory dermatoses. Chronic hand dermatoses of different and often mixed genesis represent a great problem in clinical practice. We have performed a prospective, open study with patients suffering from psoriasis, eczema or pustulosis of the hands treated with hand UVB-TL01. Thirty patients (23 women and 7 men) were included, mean age 43 years (range 25-68 years). The psoriatics were the best responders, as 9 of 11 were improved or much improved after 20-38 treatments, compared to 11 of 16 patients with eczema (11-31 treatments). One of three patients with palmar pustulosis experienced improvement after 23-27 treatments. UVB-TL01 seems to be helpful in the majority of patients with chronic inflammatory diseases of the hands. More extensive studies with regard to proper dosing and length of treatment period ought to be carried out. PMID- 15339077 TI - Failure to detect clonality in eosinophilic pustular folliculitis with follicular mucinosis. AB - Eosinophilic pustular folliculitis is characterized by an eosinophil-rich inflammatory follicular and perifollicular infiltrate primarily centred at the level of the follicular isthmus and sebaceous duct. Follicular mucinosis has been observed in lesions of eosinophilic pustular folliculitis. Clinical and histological features of eosinophilic pustular folliculitis with follicular mucinosis and alopecia mucinosa are very similar. Alopecia mucinosa may be a clonal T-cell dermatosis. A monoclonal re-arrangement of the T-cell receptor gene was detected in about half of the cases in alopecia mucinosa. To investigate T cell clonality in a series of eosinophilic pustular folliculitis with follicular mucinosis, we performed heteroduplex analysis of re-arranged T-cell receptor gamma gene in seven cases of eosinophilic pustular folliculitis with follicular mucinosis. All cases were negative for heteroduplex-PCR analysis. The failure to demonstrate clonality may be consistent with a reactive nature of eosinophilic pustular folliculitis with follicular mucinosis. PMID- 15339078 TI - Exacerbation of psoriasis after treatment with an EGFR tyrosine kinase inhibitor. PMID- 15339079 TI - A novel mutation (c. 1072_1074delGAG) in the alpha-galactosidase gene of a Taiwanese family with Fabry disease. PMID- 15339080 TI - A case of cutaneous pseudolymphoma associated with silicone injection. PMID- 15339081 TI - Drug-induced blaschkitis. PMID- 15339082 TI - Juvenile pemphigus vegetans of the glans penis. PMID- 15339083 TI - D-penicillamine-induced pemphigus vulgaris in a patient with scleroderma rheumatoid arthritis overlap syndrome. PMID- 15339084 TI - Cryptococcosis: long-lasting presence of fungi after successful treatment. PMID- 15339085 TI - Cutaneous lesions in Blastocystis hominis infection. PMID- 15339086 TI - Spitz naevus of the glans penis: an unusual location. PMID- 15339087 TI - Nodal peripheral T-cell lymphoma with secondary granulomatous cutaneous involvement. PMID- 15339088 TI - Non-Hodgkin's lymphoma following treatment of atopic eczema with cyclosporin A. PMID- 15339089 TI - 'Watering-can penis' in pseudoepitheliomatous, keratotic and micaceous balanitis. PMID- 15339090 TI - Orificial tuberculosis: presenting as a refractory perianal ulcer. PMID- 15339091 TI - Ringworm of the scalp among immigrants in Finland. PMID- 15339092 TI - Dapsone hypersensitivity or viral exanthem? PMID- 15339094 TI - [Project on the delinquency law, it's time once again to react!]. PMID- 15339097 TI - [Towards a complementary education of psychiatric nurses]. PMID- 15339099 TI - [Let's talk about love]. PMID- 15339100 TI - [The false love of transference, or the risky love in the analytic situation]. PMID- 15339101 TI - [The face of love in psychiatry]. PMID- 15339102 TI - [They love each other...they stay! I love him...I will leave]. PMID- 15339103 TI - [Erotomania]. PMID- 15339104 TI - [Management of sexual aggressors, when care is an obligation]. PMID- 15339105 TI - [When the interview is by a nurse]. PMID- 15339106 TI - [Observance, care contract and social contract]. PMID- 15339107 TI - [Nursing practices and theoretical models]. PMID- 15339108 TI - [1/5 Rogerian helping relationship]. PMID- 15339109 TI - The infertile couple: a cohort study in Duhok, Iraq. AB - From January 2000 to May 2001 causes of infertility were studied for 250 couples, 193 (77.2%) of whom had primary infertility and 57 (22.8%) had secondary infertility. Male infertility was found in 36.8% of cases. The most common cause of female infertility was ovulation disorders (41%) while tubal obstruction contributed to only 5% of cases. Only 15% of the couples had unexplained infertility. Galactorrhoea and/or increased serum prolactin was seen in 60% of cases. Ultrasonographic detection of mature follicles was consistent with the results of cervical mucus study and endometrial biopsy in 90% of cases. PMID- 15339110 TI - History of miscarriage as a risk factor for hepatitis C virus infection in pregnant Iraqi women. AB - Sera from 3491 pregnant women were screened for the presence of HCV antibodies (anti-HCV). HCV genotyping was also performed on the sera of 94 women. The overall anti-HCV seroprevalence was 3.21%. Anti-HCV seroprevalence was significantly positively correlated with the number of miscarriages. Miscarriage was a significant risk factor for the acquisition of HCV infection from the first miscarriage up to the fifth, the risk increasing with increasing number of miscarriages. A higher proportion of women with a history of miscarriage harboured HCV-1b compared to those with no miscarriage. PMID- 15339111 TI - Malaria in pregnancy in Hodiedah, Republic of Yemen. AB - In the Republic of Yemen, Plasmodium falciparum is the predominant causative agent of malaria and is associated with adverse consequences for pregnant women and their babies. The prevalence and clinical manifestations of malaria among 500 pregnant (260) and non-pregnant (240) women were compared. Clinical examinations, laboratory investigations and a structured questionnaire were used to collect data. The prevalence of malaria was higher among pregnant women (55%) than non pregnant women (20%). Anaemia was significantly more prevalent among pregnant woman than non-pregnant women and also more prevalent in pregnant women with malaria than non-pregnant women with malaria. PMID- 15339112 TI - In utero exposure to cadmium pollution in Cairo and Giza governorates of Egypt. AB - Serum cadmium levels at delivery were measured in a consecutive sample of 100 mother-infant pairs in Egypt using venous blood from mothers and umbilical cord blood from neonates. The serum cadmium levels of mothers ranged from 0.4 to 2.2 microg/L (mean 0.73 microg/L) and of infants from 0.2 to 1.5 microg/L (mean 0.66 microg/L). Infant cadmium levels were about 70% of maternal levels in most pairs. Serum cadmium was significantly higher in mothers and babies passively exposed to tobacco smoke. Five-minute Apgar scores were negatively correlated with cord blood cadmium levels. The cadmium levels did not differ between subjects from Cairo and Giza or according to urban, suburban or rural areas. Thus, in utero exposure to cadmium was evident and wider-scale studies on its long-term effects are recommended. PMID- 15339113 TI - Nutritional status of Libyan children in 2000 compared with 1979. AB - This cross-sectional study examined the growth and nutritional status of Libyan preschool children and the socioeconomic status of their parents in 2000 and compared the results with those of a similar study conducted in 1979. The investigation involved 1614 healthy children of both sexes under 5 years of age in two regions of the Libyan Arab Jamahiriya. Simple random sampling and cluster sampling techniques were used to enlist participants. Body weight, height, and head and arm circumference of the children were recorded and mothers were interviewed about their child's health and the socioeconomic status of the family. Significant improvements in nutritional status since 1979 were found that could be attributable to factors such as socioeconomic development, longer breastfeeding, greater availability of high energy foods, supplementation with semi-solid or solid foods and heavy government subsidy of basic foods. PMID- 15339114 TI - Nutritional stunting in Egypt: which nutrient is responsible? AB - We looked at the relationship between linear growth retardation and deficiencies of certain nutrients in Egyptian children. A group of 107 stunted children aged between 10 and 18 years were subjected to history-taking, physical examination and laboratory investigations. Selected cases were referred to radiology for assessment of bone age. Thirty-nine children were enrolled as controls. Serum haemoglobin, alpha-tocopherol, retinol and magnesium levels were significantly decreased in stunted children compared with the controls. Serum zinc levels were also lower in the stunted group but not significantly so. We conclude that several nutrient deficiencies occur simultaneously in stunted children and all of them may be responsible for stunting. PMID- 15339115 TI - Evaluation of erythropoiesis in protein energy malnutrition. AB - This study evaluated erythropoiesis in 50 infants hospitalized with protein energy malnutrition and in 50 control infants. The red cell count, mean corpuscular haemoglobin and reticulocyte index were significantly lower, while the white blood cell count, median corpuscular fragility and red cell distribution width were significantly higher on admission than in controls. Total serum protein, albumin, fasting blood glucose, and serum folate were significantly lower on admission than in controls. Serum ferritin was significantly higher and total iron-binding capacity was significantly lower on discharge compared to controls. The serum erythropoietin was significantly higher on admission and discharge than in controls. The anaemia of protein energy malnutrition is due to mixed deficiencies resulting in ineffective erythropoiesis despite an increased level of erythropoietin. PMID- 15339116 TI - Impact of nutritional rehabilitation on enzymatic antioxidant levels in protein energy malnutrition. AB - To assess the role of enzymatic antioxidants in the pathogenesis of protein energy malnutrition (PEM) and the effect of nutritional rehabilitation, we studied 30 infants with PEM (mean age 10.63 +/- 4.39 months: 10 marasmic; 8 with kwashiorkor; 12 with marasmic kwashiorkor) and 15 controls. All underwent clinical examination and laboratory investigations, including superoxide dismutase (SOD) and glutathione peroxidase (GPx) estimation before and after nutrition rehabilitation. SOD and GPx were significantly lower in all malnourished infants compared to controls, and significantly increased after nutritional rehabilitation. These significant correlations suggest that antioxidants could be introduced during PEM nutritional rehabilitation to decrease morbidity and mortality. PMID- 15339117 TI - Suitability of soluble transferrin receptor for the clinical diagnosis of different types of anaemia in children. AB - We evaluated the ability of serum transferrin receptor (sTFR) to identify different types of anaemia in children. Thus 150 Egyptian children suffering from anaemia (iron deficiency anaemia, anaemia of chronic disease and beta thalassaemia) were enrolled, together with 50 controls. There was a significant increase in the mean levels of sTFR in the groups with iron deficiency anaemia and thalassaemia, and a significant decrease in mean sTFR levels in the group with anaemia of chronic disease. Serum ferritin levels were significantly higher in all patient groups except the group with iron deficiency anaemia. There were also significant correlations between the sTFR and sTFR/log ferritin ratio (sTFR F index) and different indices of iron status and of erythropoiesis. The sTFR-F index could be used as a diagnostic or screening tool for iron deficiency anaemia, anaemia of chronic disease and thalassaemia. PMID- 15339118 TI - Epidemiology of children's cycling injuries in Ahwaz, Islamic Republic of Iran. AB - A one-year study was carried out in Ahwaz, Islamic Republic of Iran to investigate the epidemiology of children's cycling injuries. Children 15 years old or younger taken to the accident and emergency (A&E) departments of all urban hospitals due to cycling injury were investigated, and their parents interviewed by questionnaire. During the study, 1079 children presented to A&E departments with cycling injuries (78.8% boys and 21.2% girls), representing 4.9% of all children presenting with injuries. The most frequent injury sites were the street (51.9%) and the home (42.3%), and the head was the part of the body most frequently injured (55.0%). None of the injured children was using a cycling helmet at the time of injury. This study suggests mandatory helmet use, safety regulations and educational programmes for bicycle riders should be established in our community PMID- 15339119 TI - Atraumatic restorative treatment and glass ionomer sealants in Tunisian children: survival after 3 years. AB - We evaluated the survival rates of atraumatic restorative treatment restorations and of glass ionomer sealants after 3 years of usage in primary and permanent teeth in Tunisian children aged 3-15 years attending rural schools, and assessed the presence or absence of dental caries in the restored teeth. After an initial survey of 1949 children, 242 were selected and agreed to undergo treatment. The loss to follow-up after 3 years was about 40%. Of those evaluated, 45.73% of one surface ART restorations in permanent teeth had survived, 54.96% of one-surface sealants in permanent teeth had survived and 27.85% of one-surface ART restorations in primary teeth had survived. Caries was found in only 19 teeth after 3 years. PMID- 15339120 TI - Reasons for delay in the management of patients with pulmonary tuberculosis. AB - The study assessed reasons for delay between patient's first symptoms of tuberculosis and initiation of therapy. Fifty newly diagnosed cases of pulmonary tuberculosis admitted to the NRITLD in Teheran were studied. Mean patient delay before consulting a physician was 12.5 +/- 10 days, significantly higher among men than women. Mean delay until the physicians' diagnosis was 93 +/- 80 days, significantly higher for women than for men. Almost no treatment delay was observed (mean 4 +/- 4 days after diagnosis had been confirmed). The major delay was the time taken by physicians to diagnose tuberculosis in symptomatic patients. An active and effective national tuberculosis programme is needed in the Islamic Republic of Iran, with integration of the programme in medical school curricula and in continuing professional training. PMID- 15339121 TI - Tuberculous meningoencephalitis in Baghdad, 1993-99: a clinical study of 224 cases. AB - This study examined all cases of tuberculous meningoencephalitis admitted to Ibn El-Khateeb Hospital for Infectious Diseases in Baghdad from 1993 to 1999. The diagnosis was based on the patient's history, clinical findings, cerebrospinal fluid analysis, microbial isolation and response to antitubercular drug therapy. The findings for cases diagnosed with tuberculous meningoencephalitis were compared with other types of meningitis. There were 224 cases comprising 5% of the total number of meningitis cases admitted during this period. The male to female ratio was 1.4:1. The age range was from 6 months to 72 years and the overall case fatality rate was 21%. PMID- 15339122 TI - Characterization of Leishmania parasites isolated from provinces of the Islamic Republic of Iran. AB - Leishmania parasites isolated in the Islamic of Iran were studied by a random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). Of 82 isolates, 80 were from cutaneous lesions, 1 from a human throat lesion and 1 from a dog. Of these, 42 isolates were L. tropica, 36 were L. major and 2 were L. infantum. There were 2 unidentified isolates (from the throat lesion and a cutaneous lesion) and these demonstrated 52% and 48% similarity with L. tropica and L. infantum. Both L. tropica and L. major were isolated from four provinces indicating a recent change in the epidemiology of cutaneous leishmaniasis. L. tropica was isolated from three provinces; L. major from one province. L. infantum was isolated from a human cutaneous lesion and from a dog in Bushehr province. PMID- 15339123 TI - Intestinal parasites, including Cryptosporidium species, in Iraqi patients with sickle-cell anaemia. AB - Stool samples were obtained from individuals admitted to three hospitals in Basra during November 1997-May 1998. Of 40 patients with sickle-cell anaemia, 25 (62.5%) had parasitic infections. In the apparently healthy comparison group, 26 of 175 individuals (14.8%) had intestinal parasitic infections, a statistically significant difference. The most common intestinal parasites isolated in the sickle-cell patients were Blastocystis hominis (36%) and Giardia lamblia (28%). The isolation rate of Cryptosporidium species in sickle-cell patients (5%) was not significantly different from that in apparently healthy individuals (1.14%). We report for the first time the isolation of Isospora belli from a sickle-cell patient in Iraq and the Mediterranean region. PMID- 15339124 TI - Prevalence of trachoma in chronic conjunctivitis, Birjand, Islamic Republic of Iran. AB - Between January 1999 and December 2000, 80 consecutive patients with chronic conjunctivitis presenting in the eye clinic of Birjand University of Medical Sciences were assessed for trachoma and chlamydial infection. Direct fluorescent antibody test was used to assess trachoma and Giemsa staining to detect chlamydial infection. Trachoma was detected in 10% of the patients and chlamydial infection detected in 6.3%. The results were similar for both sexes. Although the findings are hospital-based, the prevalence was not so high as to suggest prophylactic use of azithromycin (World Health Organization protocol) for eradication of trachoma in the Birjand population. Further epidemiological studies are recommended. PMID- 15339125 TI - Impact of schistosomiasis on quality of life and productivity of workers. AB - The effect of schistosomiasis on quality of life (QOL) and productivity of workers was examined. In a textile factory in Alexandria, Egypt, personal, occupational and sociodemographic data were collected from 172 workers with schistosomiasis and 172 workers without schistosomiasis. Several indicators of productivity and the World Health Organization QOL brief were used to determine the impact of schistosomiasis. The disease affected the general, physical and independence, psychological and spiritual, and social domains of QOL. Although the productivity score of workers with schistosomiasis did not differ significantly from the control group, they had significantly lower additional hours of work and lower total incentives/month. A significant relationship was found between severity of schistosomiasis and QOL domains and productivity indicators. PMID- 15339126 TI - Skin diseases and enzymatic antioxidant activity among workers exposed to pesticides. AB - In this study, 150 workers exposed to pesticides and 50 healthy control subjects were given clinical and dermatological examinations, patch tests, tests of liver and renal function, complete blood count, blood sugar and urinalysis. Activity of the antioxidant enzymes superoxide dismutase, glutathione peroxidase and glutathione reductase was also evaluated. Dermatological findings were positive in 78%, 76% and 54% of workers exposed to organophosphates, pyrethroids and carbamate pesticides respectively. The patch test was positive in 70% of workers exposed to pyrethroids and 64% exposed to carbamate pesticides. Liver enzyme levels were generally increased in workers while antioxidant enzyme activity was significantly decreased in all workers compared with the controls. PMID- 15339127 TI - Prevalence of known diabetes and hypertension in the Republic of Yemen. AB - The present study was undertaken to determine the prevalence of known cases of diabetes and hypertension among adults in Sana'a city. Thus 1080 persons aged 20 85 years were selected for interview using a multistage random sampling technique. The crude prevalence of known diabetes was 6.57% (95% CI: 5.2-8.2) and of known hypertension 13.5% (95% CI: 11.5-15.6). The age-standardized prevalence for the age range 30-64 years was 9.75% (95% CI: 7.55-11.95) for diabetes and 17.1% (95% CI: 15.0-19.2) for hypertension. These results provide an estimate of the prevalence of known diabetes, hypertension and related vascular events in a well-defined urban community. PMID- 15339128 TI - Assessment of a breast cancer screening programme in Shiraz, Islamic Republic of Iran. AB - A breast cancer screening programme was evaluated for approximately 10,000 women aged 35 years and older. There were 67 cases of breast cancer. Highest rates of attendance were seen among younger women (35-44 years) and middle socioeconomic groups. Lowest rates were among those aged over 65 years and low socioeconomic groups. The rate of detection by self-examination was similar to that by health personnel examination. At all stages of screening, positive findings were most common among the high socioeconomic class. Attendance decreased steadily from first to last stages of serial screening. Although mammography is the most sensitive method of detection, because of its high cost we suggest establishing breast self-examination education programmes and encouraging women to self examine. PMID- 15339129 TI - Flow cytometric DNA analysis of oral squamous cell carcinoma in Iraqi patients. AB - We studied the DNA content, DNA index and cell cycle parameters that are reliable markers for assessing the proliferative activity and aggressiveness of malignancies. Cytometric DNA analysis was performed on formalin-fixed paraffin embedded sections from 36 Iraqi patients with oral squamous cell carcinoma. The results showed that 20 of 36 cases (55.5%) were diploid, while 15 cases (41.7%) were aneuploid. Significantly higher S-phase fractions and higher DNA indices characterized aneuploid tumours. Nuclear DNA analysis as part of the evaluation of oral squamous cell carcinoma will influence the selection of appropriate treatment strategies. PMID- 15339130 TI - Causes of visual impairment and blindness among the middle-aged and elderly in northern Jordan. AB - Causes of bilateral low vision (BLLV) and bilateral blindness (BLB) were investigated among 720 outpatients aged > 45 years in Irbid, Jordan, between July 1999 and October 2000. All received full ophthalmic examinations. BLLV was defined as visual acuity < 6/18 but > 3/60 in the better eye; BLB as acuity < 3/60 in the better eye. The leading cause of BLB and BLLV was cataract. Refractive error was the second major cause of BLLV. Other common causes were diabetic retinopathy and glaucoma. Age-related macular degeneration was the single cause of BLB and BLLV for only 1.6% and 2.8% respectively. A population based survey of BLB and BLLV in the elderly, yearly vision examinations and an ophthalmic education programme are recommended. PMID- 15339131 TI - Changing use of the emergency department by the elderly in the United Arab Emirates, 1989 and 1999. AB - Changes in use by the elderly of the emergency department of a major hospital in Al-Ain in the years 1989 and 1999 were examined. All patients aged 65 years or older who attended the emergency room were included in this cross-sectional survey. Patient numbers rose from 321 in 1989 to 1347 in 1999 with no significant change in the female to male ratio of 0.78 or the mean age of the elderly attendees 72.9 +/- 7.4 years. Attendance rose 5.4 fold while non-urgent attendance rose 14.7 fold, demonstrating rising use but falling illness severity over time. PMID- 15339132 TI - Survey on adoption of measures to prevent nosocomial infection by anaesthesia personnel. AB - To assess the knowledge, attitudes and practices of anaesthesia personnel regarding infection control in hospitals, a questionnaire was distributed to anaesthesiology personnel of different educational levels in the hospitals of southern Islamic Republic of Iran. Chi-squared significance, Fisher exact and Spearman rho correlation coefficient tests were used to analyse the responses. The results suggest that measures to prevent infection transmission during anaesthesia are inadequate in our hospitals. The implementation of adequate measures to control infection was significantly associated with respondents' beliefs as to whether anaesthesia can cause infection in anaesthesia personnel and/or patients. Increasing the knowledge base of anaesthesia personnel and raising their awareness as to the risk of infection are necessary to improve infection control procedures by anaesthesia personnel. PMID- 15339133 TI - In-home drug storage and utilization habits: a Sudanese study. AB - Community drug-use habits were studied in 469 household units in different areas of Sudan. About 97.7% of the investigated families had at least one drug product stored at home. The study revealed a high rate of self-medication (46.9%), repeated use of unfinished stored drugs (55.0%), a high rate of drug exchange among families (59.3%) and poor compliance (71.2%). In Sudan there is still a great need to educate and to motivate the general public regarding the principles of rational drug use in order to safeguard health and avoid economic losses. PMID- 15339134 TI - Depleted uranium: an overview of its properties and health effects. AB - There has been much debate about the use of depleted uranium in the Gulf War and its health effects on United States and European war veterans. However, studies on the impact of this radioactive substance on the residents of the surrounding Gulf region are far from adequate. Depleted uranium introduces large quantities of radioactive material that is hazardous to biological organisms, continues to decay for millennia and is able to travel tens of kilometres in air. If depleted uranium were used in the Gulf War, its impact on the health of people in the area would have been considerable. This review of depleted uranium--its origin, properties, uses and effects on the human environment and health--aims to trigger further research on this subject. PMID- 15339135 TI - [Osteoporosis]. AB - It is well known that people, especially white people, are getting osteoporosis more often than previously thought. Until now, no direct causative factor has been determined, but genetic factors are very likely to be involved. Usually, affected individuals are initially asymptomatic while the disease process is going on, and they come to the attention of the medical profession only late when their bones are fractured as a result of a simple trauma. Also it is vital to let people know that heavy sports, at times, can be harmful. PMID- 15339136 TI - [A single window for access to the Ministry of Health studies and research in Morocco]. AB - This article highlights that there is a need for an inventory of all the health system research and studies which have been conducted by the Moroccan Ministry of Health and its partners, and that this knowledge base should be made available to the decision-makers, the researchers and even the public. This could be done using the new information and communication technology which is available at the Ministry. This will gradually lead to establishing a single-window service for access to health knowledge and know-how in the near future. PMID- 15339137 TI - Gel permeation chromatography as a combinatorial screening method: identification of highly active heteroligated phenoxyimine polymerization catalysts. AB - A number of mixed-ligand bis(phenoxyimine) titanium catalysts were screened using a pooled combinatorial approach. Gel permeation chromatography (GPC) was used to analyze polypropylene mixtures, leading to the identification of highly active catalysts. Complexes with only one ortho-fluorinated ligand are significantly more active than homoligated complexes for the syndiospecific polymerization of propylene. PMID- 15339138 TI - 2,9-Di-(2'-pyridyl)-1,10-phenanthroline: a tetradentate ligand for Ru(II). AB - The tetradentate ligand 2,9-di-(2'-pyridyl)-1,10-phenanthroline is synthesized in 62% yield by the Stille coupling of 2,9-dichloro-1,10-phenanthroline and 2-(tri-n butylstannyl)pyridine. Treatment of this ligand with RuCl3.3H2O and a 4 substituted pyridine results in the formation of a complex in which the tetradentate ligand occupies the equatorial plane and two pyridines are bound axially. The interior N-Ru-N angles vary from 76.1 degrees to 125.6 degrees , showing considerable distortion from the 90 degrees ideal. The lowest energy electronic transition is sensitive to the electronegativity of the 4-substituent on the axial pyridines, varying from 516 nm for the CF3 group to 580 nm for the NMe2. The oxidation potentials mirror this trend, spanning a range of 1.36-1.03 V, while the reduction potentials show less variation (-0.97 to -1.08 V). The complexes are nonemissive, presumably due to competitive nonradiative processes caused by distortion of the system. PMID- 15339139 TI - Reactivity of amido ligands on a dinuclear Ru(II) center: formation of imido complexes and C-N coupling reaction with alkyne. AB - Treatment of a dinuclear Ru(II) amido complex [Cp*Ru(mu2-NHPh)]2 (Cp* = eta5 C5Me5) with small organic substrates including CO, tert-butyl isocyanide, a sulfur ylide Ph2S=CH2, and diphenylacetylene resulted in an unexpected disproportionation reaction of the bridging amido ligands to produce a free amine and a series of imido-bridged diruthenium complexes [(Cp*Ru)2(mu2-L)(mu2-NPh)] (L = CO, t-BuNC, CH2). In the case of diphenylacetylene, the bridging imido ligand underwent subsequent coupling reaction with the coordinated alkyne to form an iminoalkenyl complex [(Cp*Ru)2(mu2-PhNCPhCPh)]. PMID- 15339140 TI - Coacervate formation from natural glycolipid: one acetyl group on the headgroup triggers coacervate-to-vesicle transition. AB - Coacervate (L3 phase) formation of the single component "natural" glycoliped biosurfactant, MEL-A, was observed for the first time by using an optical microscope, a confocal laser scanning microscope (CLSM), and a freeze-fracture electron microscope (FF-TEM). It was also found that only a slight decrease in spontaneous curvature resulting from the absence of one acetyl group on the headgroup induced a drastic morphological change in the 3D self-assembled structure from coacervates (L3 phase) to ordered vesicles (Lalpha phase). PMID- 15339141 TI - Recognition and stabilization of peptide alpha-helices using templatable nanoparticle receptors. AB - alpha-Helices are important structural elements in proteins. To provide a scaffold for the facial recognition of peptides, we have explored the interaction of cationic mixed monolayer protected clusters (MMPCs) with a tetra-aspartate peptide in water. In these studies, substantial enhancement of peptide helicity was observed upon addition of the MMPC. Significantly, this stabilization increased with time, demonstrating templation of the monolayer to the peptide helix. PMID- 15339142 TI - Arrays for the combinatorial exploration of cell adhesion. AB - A new method for the fabrication of arrays of self-assembled monolayers (SAMs) of alkane thiols (ATs) on gold to combinatorially assay surfaces for cell adhesion is reported. A fluorous SAM, which is both cytophobic and solvophobic, was used as the background between the array features. The resulting solvophobic background permits the application of an assembly after conjugation strategy for fabrication. SAMs containing mixtures of ATs and peptide-terminated ATs were generated. Multiple cell types demonstrated differential and specific binding to these surfaces. Additionally, pluripotent human embryonic stem cells proliferated on surfaces generated by this method. PMID- 15339143 TI - Ultrafast aggregate-to-aggregate energy transfer within self-assembled light harvesting columns of zinc phthalocyanine tetrakis(perylenediimide). AB - Light harvesting in photosynthetic antenna proteins involves a series of highly efficient ultrafast energy transfers between spectroscopically different populations of chlorophylls. Several strategies have recently been employed to mimic this natural energy transfer process, including polymers, dendrimers, and oligomeric porphyrin arrays linked by covalent bonds or by self-assembly. In all of these systems, excitation energy transfer occurs from one molecule to another, while very few of them involve energy transfer from one very strongly interacting chromophore aggregate to another such aggregate. Here we report the synthesis and characterization of a covalent zinc phthalocyanine-2,3,9,10,16,17,23,24 octacarboxytetraimide in which all four imide nitrogen atoms are substituted with N-octyl-N'-(4-aminophenyl)-1,7(3',5'di-tert-butylphenoxy)perylene-3,4:9,10 bis(dicarboximide) (ZnPcIm4-PDI4). The individual molecules self-assemble into stacked heptamers in solution as evidenced by small-angle X-ray scattering and form long fibrous structures in the solid as evidenced by TEM. The ZnPcIm4 and PDI molecules both stack in register with the same components in an adjacent covalent building block. Ultrafast energy transfer occurs with tau = 1.3 ps from the aggregated peripheral PDI chromophores to the core ZnPcIm4 chromophore aggregate. Exciton hopping between the ZnPcIm4 chromophores occurs with tau = 160 fs. PMID- 15339144 TI - A site-selective dual anchoring strategy for artificial metalloprotein design. AB - Introducing nonnative metal ions or metal-containing prosthetic groups into a protein can dramatically expand the repertoire of its functionalities and thus its range of applications. Particularly challenging is the control of substrate binding and thus reaction selectivity such as enantioselectivity. To meet this challenge, both non-covalent and single-point attachments of metal complexes have been demonstrated previously. Since the protein template did not evolve to bind artificial metal complexes tightly in a single conformation, efforts to restrict conformational freedom by modifying the metal complexes and/or the protein are required to achieve high enantioselectivity using the above two strategies. Here we report a novel site-selective dual anchoring (two-point covalent attachment) strategy to introduce an achiral manganese salen complex (Mn(salen)), into apo sperm whale myoglobin (Mb) with bioconjugation yield close to 100%. The enantioselective excess increases from 0.3% for non-covalent, to 12.3% for single point, and to 51.3% for dual anchoring attachments. The dual anchoring method has the advantage of restricting the conformational freedom of the metal complex in the protein and can be generally applied to protein incorporation of other metal complexes with minimal structural modification to either the metal complex or the protein. PMID- 15339145 TI - DNA-PEG-DNA triblock macromolecules for reagentless DNA detection. AB - The sandwich assay is the most common design for electrochemical DNA sensors. This assay consists of three individual DNA components: an immobilized capture strand, a target strand, and a probe strand containing a redox-active reporter group. We report a simplified DNA assay where two strands of ssDNA, the capture and probe strands, are linked together via a flexible poly(ethylene glycol) (PEG) spacer forming an ABA triblock macromolecule. We have developed an electrochemical assay where the detection signal arises as a consequence of a large structural change induced upon hybridization with target DNA. In this system, the DNA-PEG-DNA macromolecule folds or wraps around the target DNA, bringing the ferrocene probe in close proximity to the electrode, affording an electrochemical response. PMID- 15339146 TI - Stereocontrol within confined spaces: enantioselective photooxidation of enecarbamates inside zeolite supercages. AB - Dye-exchanged Y zeolite is shown to be an effective medium to control the stereoselectivity in the photooxygenation of chiral oxazolidinone-functionalized Z/E-1 enecarbamates. An enantioselectivity (ee) as high as 80% was observed in the methyldesoxybenzoin (MDB) product, obtained in the methylene-blue-exchanged NaY zeolite at room temperature. The efficacy of the asymmetric induction in the MDB product depends on the Z/E geometry of the alkene, the Z-isomer being more effective than the corresponding E-isomer. The stereoselectivity is rationalized in terms of conformational effects through cationic interactions between the zeolite and the substrate. PMID- 15339147 TI - A 3.5-nm coordination nanotube. AB - A 3.5-nm coordination nanotube was self-assembled from 12 Pd(II) ions and four tape-shaped ligands with the aid of a 3.0-nm template. This tubular structure was unambiguously determined by the single-crystal X-ray analysis. Being cooperatively sustained by 24 Pd(II)-pyridine interactions, the framework was kinetically stable even when the template was removed. The empty tube was, of course, capable of binding other molecules in the cavity. PMID- 15339148 TI - Experimental and theoretical determination of nucleic acid magnetic susceptibility: importance for the study of dynamics by field-induced residual dipolar couplings. AB - Magnetic susceptibility anisotropies (Deltachi) are determined for the nucleic acid bases on the basis of a combined NMR spectroscopic and DFT study. Magnetic field-induced residual dipolar couplings (RDC) measured for the Dickerson dodecamer allow for the determination of a precise value of Deltachi for this DNA. A tensor summation employing the results of DFT calculations of the individual base magnetic susceptibility tensors and the known structure of the dodecamer provides a value of Deltachi which is in excellent agreement with experiment. The DFT results are shown to provide superior agreement with experiment for the dodecamer and a protein-DNA complex when compared with other sets of values in common use in the literature. The marked discrepancy between our Deltachi values and existing sets of values has a direct and significant impact on the interpretation of field-induced anisotropic interactions in terms of biomolecular structure and dynamics. PMID- 15339149 TI - Direct patterning of copper on polyimide using ion exchangeable surface templates generated by site-selective surface modification. AB - We demonstrate site-selective chemical surface modification by dispensing potassium hydroxide solution onto polyimide, which confines source metallic ions that can subsequently be used in resist- and mask-free fabrication of copper circuit patterns. Metallization can be achieved by a wet chemical method, providing control over metal/polymer interfacial structures. Because the approach is compatible with other existing printing technologies and much simpler than conventional lithography-based methods, we propose that the present surface template method may be of general application in fabrication of metallized polymers as well as in development of integrated circuits with a variety of electronic circuit elements. PMID- 15339150 TI - Crystallographic structures of two bisphosphonate:1-deoxyxylulose-5-phosphate reductoisomerase complexes. AB - We have obtained the single-crystal X-ray crystallographic structures of the bisphosphonates [(1-isoquinolinylamino)methylene]-1,1-bisphosphonate and [[(5 chloro-2-pyridinyl)amino]methylene]-1,1-bisphosphonate, bound to the enzyme 1 deoxyxylulose-5-phosphate reductoisomerase (DXR, EC 1.1.1.267, also known as 2-C methyl-d-erythritol-4-phosphate synthase), an important target for the development of antimalarial drugs. Our results indicate that both bisphosphonates bind into the fosmidomycin binding site. The aromatic groups are in a shallow hydrophobic pocket, and the phosphonate groups are involved in electrostatic interactions with Mg2+ or a cluster of carboxylic acid groups and lysine while the fosmidomycin phosphonate-binding site is occupied by a sulfate ion (as also observed in the DXR/NADP+ structure). The availability of these two new crystal structures opens up the possibility of the further development of bisphosphonates and related systems as DXR inhibitors and, potentially, as antiinfective agents. PMID- 15339151 TI - Dye-activated hybrid organic/inorganic mesostructured titania waveguides. AB - A room-temperature synthesis route for the fabrication of a new type of hybrid organic/inorganic mesostructured material based on titania instead of silica as the inorganic component has been developed. This approach enables facile processing of the titania/block copolymer surfactant precursor solution into optically activated, transparent, and crack-free fibers and planar waveguides with adjustable thickness. Stabilization of the structures occurs upon solvent evaporation by formation of a solid, glasslike material without heat treatment. These dye-activated, high refractive index, titania-based composites show efficient waveguiding and mirrorless lasing at low thresholds without the need of an ultralow refractive index support layer. PMID- 15339152 TI - Stabilization of yeast cytochrome C covalently immobilized on fused silica surfaces. AB - The surface-enhanced conformational stability of yeast cytochrome c (YCC) covalently immobilized on a fused silica prism with heterobifunctional cross linkers has been studied by attenuated total reflection absorption spectroscopy using the Soret band of the heme prosthetic group as a probe. Comparison of the results to those of horse cytochrome c physisorbed on the same substrate as well as to the corresponding proteins in solution indicates that the surface plays a significant role in stabilizing the native conformation of the surface-bound YCC. Unfolding to extended configurations was so hindered that the native conformation of the covalently immobilized protein is essentially unaffected by the presence of denaturants such as methanol and 1-propanol. PMID- 15339153 TI - Edge spreading lithography and its application to the fabrication of mesoscopic gold and silver rings. AB - A new form of edge lithography, edge spreading lithography (ESL), has been demonstrated and applied to the formation of coinage metal rings. In this process, alkanethiols are delivered from a flat PDMS stamp to the surface of a metal film through a two-dimensional array of spherical silica colloids. The thiols further spread on the metal surface, forming highly ordered SAMs in the form of a ring pattern. Following lift-off of beads, the pattern in the SAMs can be transferred into the metal film through wet chemical etching, with SAMs serving as the resist. The dimensions of the rings can be readily controlled by several parameters such as the beads diameter, the concentration of the thiol solution, and the contact time between the stamp and the silica beads. PMID- 15339154 TI - Discrete nanostructures of quantum dots/Au with DNA. AB - Nanostructures of colloidal CdSe/ZnS core/shell quantum dots (QDs) surrounded by a discrete number of Au nanoparticles were generated via DNA hybridization and purified by gel electrophoresis. Statistics from TEM analysis showed a high yield of designed structures. The distance between Au particles and QD, the number of Au around the central QD, and the size of Au and QD can be adjusted. Rationally designed structures such as these hold great promise for researching the physical interactions between semiconductor and Au nanoparticles and for developing more efficient nanoprobes. PMID- 15339155 TI - A synthetic DNA walker for molecular transport. AB - Inspired by kinesin movement along a microtubule, we demonstrate a processive bipedal DNA walker. Powered by externally controlled DNA fuel strands, the walker locomotes with a 5 nm stride by advancing the trailing foot to the lead at each step. Real-time monitoring of specific bidirectional walker movement is achieved via multiplexed fluorescence quenching. PMID- 15339156 TI - Electron transfer between cytochrome c and cytochome c peroxidase in single crystals. AB - Cytochrome c (Cc) and cytochrome c peroxidase (CcP) form an important redox pair for understanding interprotein electron transfer (ET). Measurements of ET rates from photoexcited CcP substituted with Zn porphyrin to either yeast Fe(III)Cc or horse Fe(III)Cc in crystals reveal that the molecular associations found in the respective crystal structures determine solution reactivity. Similar forward rates for yeast isozyme-1 Cc (yCc) and yCc homologue horse Cc (hCc), despite different orientations relative to CcP, suggest small-amplitude conformational gating of ET even in the crystalline state; faster back ET in the yCc compared to the hCc complex agrees with the relative coupling between redox sites predicted by the structures. PMID- 15339157 TI - Dramatic effects of boryl substituents on thermal ring-closing reaction of vinylallenes. AB - The unidirectional thermal ring-closing reaction of cis-4-phenyl-5-borylpenta 1,2,4-triene giving 4-boryl-3-methylene-1-phenylcyclobutene proceeded significantly faster than that of the trans-isomer. The large rate difference between the cis- and trans-stereoisomers is ascribed to electronic participation of the vacant boron p orbital in the transition state SHOMO. PMID- 15339158 TI - Formation of a hydrogen-bonded receptor assembly in niosomal membranes. AB - The formation of a hydrogen-bonded receptor in niosomal membranes is reported in this communication. The niosomes prepared from non-ionic surfactants C12EO3 and C14EO3 provide a stable system that allows the self-assembly of hydrogen-bonded receptors to occur in contact with aqueous environments. We demonstrate here by CD, TEM and Confocal Microscopy that the incorporation of the building blocks, calix[4]dimelamine 1 and PerBAR, within the niosomal membranes led to the formation of well-defined hydrogen-bonded assembly 13.(PerBAR)6 within these membranes. PMID- 15339159 TI - Nanosizing drug particles in supercritical fluid processing. AB - The supercritical fluid-processing technique, rapid expansion of a supercritical solution into a liquid solvent (RESOLV), was applied to the nanosizing of water insoluble drug particles. Selected for demonstration were antiinflammatory drugs Ibuprofen and Naproxen, for which CO2 and CO2-cosolvent systems were used. The RESOLV process produces exclusively nanoscale (less than 100 nm) Ibuprofen and Naproxen particles suspended in aqueous solutions, and the aqueous suspensions of the drug nanoparticles are protected from particle agglomeration and precipitation by using common polymeric and oligomeric stabilization agents. PMID- 15339160 TI - Dynamic nuclear polarization with biradicals. AB - Dynamic nuclear polarization (DNP) experiments in rotating solids have been performed for the first time using biradicals rather than monomeric paramagnetic centers as polarizing agents. Specifically, two TEMPO radicals were tethered with a poly(ethylene glycol) chain of variable length where the number of glycol units was 2, 3, or 4. NMR experiments show that the signal observed in DNP experiments is approximately inversely proportional to the length of the chain. Thus, the shorter chain with larger electron dipolar couplings yields larger enhancements. The size of the enhancement is a factor of 4 larger than obtained with the identical concentration of monomeric nitroxide radicals achieving a value of approximately 175 for the n = 2 chain. PMID- 15339161 TI - Complete control of the chemoselectivity in catalytic carbene transfer reactions from ethyl diazoacetate: an N-heterocyclic carbene-Cu system that suppresses diazo coupling. AB - The complex IPrCuCl (1) catalyzes the transfer of the :CHCO2Et group from ethyl diazoacetate (EDA) to unsaturated and saturated substrates (olefins, amine, alcohols) with very high yields. In the absence of substrate, the complex 1 does not react with EDA to give the diazo coupling products (fumarate and maleate), a rare example in the field of metal-catalyzed diazocompounds decomposition. PMID- 15339162 TI - Fast cleavage kinetics of a natural hammerhead ribozyme. AB - The hammerhead ribozyme is a small RNA motif that catalyzes the cleavage and ligation of RNA. The well-studied minimal hammerhead motif is inactive under physiological conditions and requires high Mg(2+) concentrations for efficient cleavage. In contrast, natural hammerheads are active under physiological conditions and contain motifs outside the catalytic core that lower the requirement for Mg(2+). Single-turnover kinetics were used here to characterize the Mg(2+) and pH dependence for cleavage of a trans-cleaving construct of the Schistosoma mansoni natural hammerhead ribozyme. Compared to the minimal hammerhead motif, the natural Schistosoma ribozyme requires 100-fold less Mg(2+) to achieve a cleavage rate of 1 min(-1). The improved catalysis results from tertiary interactions between loops in stems I and II and likely arises from increasing the population of the active conformation. Under optimum pH and Mg(2+) conditions this ribozyme cleaves at over 870 min(-1) at 25 degrees C, further demonstrating the impressive catalytic power of this ribozyme. PMID- 15339163 TI - Conical-nanotube ion-current rectifiers: the role of surface charge. AB - Ion channels are protein pores that span cell membranes and open and close in response to stimuli like changes in the transmembrane potential, binding of a ligand, or mechanical stress. When open, ions pass through the pore, and hence across the cell membrane, and when closed, ion-transport is precluded. Hence, these channels are nanodevices that have a current-rectification function. There is intense research effort aimed at understanding the molecular-level mechanism for this function. One approach for elucidating the mechanism is to construct a simple abiotic system that mimics this function and to use the mechanistic details of this mimic as a guide to understand the more complex biological channel. We describe here such an abiotic mimic: a synthetic membrane that contains a single conical gold nanotube. The advantage of this mimic is that the surface charge and chemistry of the nanotube wall can be varied, at will, by judicious choice of electrolyte or by thiol chemisorption. This has allowed us to make conical Au nanotubes that rectify the ion current and, just as importantly, to definitively elucidate the mechanism of this function. PMID- 15339164 TI - Low-temperature stability and high-temperature reactivity of iron-based core shell nanoparticles. AB - Iron-based core-shell nanoparticles have been prepared that exhibit low temperature stability and high-temperature reactivity toward oxygen in solution. The concentration of oxygen was determined from the fluorescence decay of pyrene. At low temperatures (<110 degrees C), the decays are short, indicating oxygen in solution. At higher temperatures (>110 degrees C), the decays become long and are consistent with no oxygen in solution. The change is abrupt, occurring over a narrow temperature range, and reproducible. PMID- 15339165 TI - Single-crystal nanowires of platinum can be synthesized by controlling the reaction rate of a polyol process. AB - Platinum nanowires of approximately 100 nm in length and approximately 5 nm in diameter have been synthesized by reducing H(2)PtCl(6) with ethylene glycol in the presence of poly(vinyl pyrrolidone) (PVP) and a trace amount of Fe(3+) or Fe(2+). The wires were generated at the final stage of the synthesis, which involved the formation of several intermediate species. The Fe(3+) or Fe(2+) ions had dual functions in the synthesis: they induced aggregation of Pt nanoparticles into larger structures that served as the nucleation sites, and they greatly reduced the reaction rate and supersaturation level to induce anisotropic growth. The reaction mechanism was studied by X-ray photoelectron spectroscopy (XPS) and UV-vis spectral analysis. The Pt nanowires could be readily separated from the surfaces of the agglomerates by sonication and obtained as pure samples by centrifugation. PMID- 15339166 TI - Transbilayer complementarity of phospholipids. A look beyond the fluid mosaic model. AB - Lipid-lipid interactions across a phospholipid bilayer were probed by measuring the nearest-neighbor preferences of exchangeable phospholipid monomers derived from 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE) and 1,2-distearoyl sn-glycero-3-phosphoethanolamine (DSPE) in the presence of nonexchangeable DMPE- or DSPE-based dimers. Each of these permanent dimers promoted homophospholipid association to the same extent, whereas the corresponding nonexchangeable monomers were without effect. These results support a model in which the longer phospholipids in one monolayer leaflet preferentially associate with shorter ones in the adjoining monolayer. Such transbilayer complementarity is likely to play an important role in stabilizing biological membranes and also in promoting a compositional interdependence of their two lipid leaflets. PMID- 15339167 TI - Catalytic isomerization of 1,5-enynes to bicyclo[3.1.0]hexenes. AB - The cycloisomerization of 1,5-enynes catalyzed by cationic triphenylphosphinegold(I) complexes produces bicyclo[3.1.0]hexenes. Substitution at all positions of the 1,5-enyne is tolerated, leading to a wide range of bicyclo[3.1.0]hexane structures, including those containing quaternary carbons. Substrates containing a 1,2-disubstituted olefin undergo stereospecific cycloisomerization (cis-olefin produces cis-cyclopropane, and trans-olefin gives trans-cyclopropane). Additionally, enantioenriched bicyclo[3.1.0]hexenes can be obtained from the gold(I)-catalyzed cycloisomerization of enantioenriched 1,5 enynes with excellent chirality transfer. The preparation of tricyclic systems is accomplished through a gold(I)-catalyzed tandem cycloisomerization-ring enlargement reaction. PMID- 15339168 TI - Catalytic coupling of terminal alkynes with isonitriles promoted by organoactinide complexes. AB - The coupling reaction of terminal alkynes and tert-butylisonitrile to yield substituted alpha,beta-acetylenic aldimines is catalyzed by the organoactinide neutral complexes Cp*2AnMe2 (Cp* = C5Me5, An = Th, U) and the cationic complex [(Et2N)3U][BPh4]. The reaction proceeds by a 1,1-insertion of the isonitrile into the metal-acetylide bond. Additional insertion products can be obtained by altering the catalyst and the reactant ratios. A plausible mechanism for the catalytic reaction is presented, in addition to the crystal structure of Cp*2UMe2 PMID- 15339169 TI - Highly diastereoselective and enantioselective C-H functionalization of 1,2 dihydronaphthalenes: a combined C-H activation/Cope rearrangement followed by a retro-Cope rearrangement. AB - The Rh2(S-DOSP)4-catalyzed reaction of vinyldiazoacetates with dihydronaphthalenes results in a highly enantioselective (91-99.6% ee) and diastereoselective (>98% de) C-H functionalization. The apparent intermolecular C H insertion was demonstrated to be a combined C-H activation/Cope rearrangement followed by a retro-Cope rearrangement. PMID- 15339170 TI - Atomic level control over surface species via a molecular precursor approach: isolated Cu(I) sites and Cu nanoparticles supported on mesoporous silica. AB - A nonaqueous molecular precursor grafting approach was employed for the generation of well-defined surface structures featuring Cu on a mesoporous silica support. X-ray absorption measurements (XANES and EXAFS) were used to determine that [CuOSi(OtBu)3]4 provided 100% isolated Cu(I) sites upon grafting (without thermal treatment), whereas [CuOtBu]4 gave isolated species with most of the original Cu-O-Cu linkages intact, but in a more relaxed straight chain form. Upon heating under inert conditions, the vast majority of Cu in the materials from [CuOSi(OtBu)3]4 remained as isolated Cu(I) sites (up to 88% isolation), with significant stabilization provided from the -OSi(OtBu)3 ligands. In stark contrast, approximately 100% of the Cu in the materials generated from [CuOtBu]4 was readily reduced upon heating, forming isolated Cu metal particles with an average diameter of 0.7 nm. PMID- 15339171 TI - Bismercaptoethanediazacyclooctane as a N2S2 chelating agent and Cys-X-Cys mimic for Fe(NO) and Fe(NO)2. AB - The N-protonated bismercaptoethanediazacyclooctane serves as a bidentate dithiolate ligand to oxidized Fe(NO)(2) of Enemark-Feltam notation, E-F [Fe(NO)(2)],(9) mimicking Cys-X-Cys binding of Fe(NO)(2) to proteins or thio biomolecules. The neutral compound is characterized by the well-known g = 2.03 EPR signal which is a hallmark of dinitrosyl iron complexes, DNIC's. The Fe(NO)(2) unit can be removed from the chelate by excess PhS(-), producing (PhS)(2)Fe(NO)(2)(-). Transfer of NO from Fe(H(+)bme-daco)(NO)(2) (nu(NO) = 1740, 1696 cm(-)(1)) to Fe(II) of [(bme-daco)Fe](2) yields the five-coordinate, square pyramidal N(2)S(2)Fe(NO) (nu(NO) = 1649 cm(-)(1)), where NO is in the apical position. Its isotropic EPR signal at g = 2.05 is consistent with E-F [Fe(NO)](7) formulation. In excess NO, Roussin's red ester-type molecules are formed as dinuclear or tetranuclear species, [(micro-SRS)[Fe(2)(NO)(4)]](n)() (n =1, 2). These well-characterized molecules furnish reference points for positions and patterns in nu(NO) vibrational spectroscopy expected to be useful for in vivo studies of NO degradation of iron-sulfur clusters in ferredoxins. PMID- 15339172 TI - Protein surface-assisted enhancement in the binding affinity of an inhibitor for recombinant human carbonic anhydrase-II. AB - We elaborate on a novel strategy for enhancing the binding affinity of an active site directed inhibitor by attaching a tether group, designed to interact with the surface-exposed histidine residue(s) of enzymes. In this approach, we have utilized the recombinant form of human carbonic anhydrase-II (hCA-II) as the enzyme source and benzenesulfonamide and its derivatives as inhibitors. The steady-state kinetic and the ligand binding data revealed that the attachment of iminodiacetate (IDA)-Cu(2+) to benzenesulfonamide (via a triethylene glycol spacer) enhanced its binding affinity for hCA-II by about 40-fold. No energetic contribution of either IDA or triethylene glycol spacer was found (at least in the ground state of the enzyme-inhibitor complex) when Cu(2+) was stripped off from the tether group-conjugated sulfonamide derivative. Arguments are presented that the overall strategy of enhancing the binding affinities of known inhibitors by attaching the IDA-Cu(2+) groups to interact with the surface-exposed histidine residues will find a general application in designing the isozyme-specific inhibitors as potential drugs. PMID- 15339173 TI - O-selectivity and utility of phosphorylation mediated by phosphite triester intermediates in the N-unprotected phosphoramidite method. AB - Previously, O-selective phosphorylation on polymer supports in the N-unprotected phosphoramidite method could not be carried out because the amino groups of dA and dC have high reactivity toward tervalent phosphorus(III)-type phosphitylating reagents. In this paper, we developed a new coupling strategy named the "activated phosphite method" in which the phosphitylation is mediated by phosphite triester intermediates 1. Application of 1-hydroxybenzotriazole as the promoter to the solid-phase synthesis resulted in excellent O-selectivity of more than 99.7%. This O-selectivity was explained by the frontier molecular orbital interactions between the reactive intermediates and the nucleophiles such as the amino or hydroxyl groups of nucleosides. Furthermore, longer oligonucleotides were synthesized not only by a manual operation but also by a DNA synthesizer. The utility of our new method was demonstrated by the successful synthesis of a base-labile modified oligodeoxyribonucleotide having 4-N-acetyldeoxycytidine residues. Finally, DNA 20-mers containing dA or dC could be synthesized in good yields by use of a combined reagent of 6-trifluoromethyl-1-hydroxybenzotriazole and benzimidazolium triflate. PMID- 15339174 TI - Mechanism and ligand-transfer selectivity of 1,2-addition of organozincate complexes to aldehyde. AB - Density functional theory (DFT) calculations have been performed to investigate the origin of the reactivity and ligand-transfer selectivity of organozincates in the 1,2-addition to carbonyl compounds. Examination of the addition of Me(3)ZnLi to formaldehyde as compared with that of Me(2)Zn showed that the addition reaction is facilitated by the push-pull synergy of the Lewis acidic Li atom and the negatively charged Me(3)Zn moiety. This analysis then provided an answer to the mechanistic question about the experimentally established ligand-transfer selectivity in the 1,2-addition of heteroleptic organozincate Me(2)Zn(X)Li (X = H , R(2)N-, and R(3)Si-). The addition of these heteroleptic zincate compounds results in selective transfer of H, R(2)N, and R(3)Si groups owing to the favorable orbital interaction between these groups and the carbonyl pi-system. The addition reaction of the zincate compounds conforms to the mechanistic framework of the conventional nucleophilic reaction, such as the addition reaction of MeLi dimer, and is different from the reaction of organocuprates, where oxidation/reduction of the copper atom is involved. PMID- 15339175 TI - X-ray crystal structures of a benzonorbornenyl cation and of a protonated benzonorbornenol. AB - The crystal structure of the 9-methylbenzonorbornenyl cation Me-1+ shows a relatively strong interaction between the sp(2)-hybridized carbon atom C9 and the aromatic ring (C4a-C9 identical with C8a-C9 = 1.897(10) A). The anion Sb(2)F(11)( ) is refined as rotationally disordered along the Sb...Sb axis. In sharp contrast to the findings about Me-1+, the protonated anti-benzonorbornenol 5+ is essentially an oxonium ion with only weak interaction between the C9 bridge and the aromatic ring despite the fact that it is already a positively charged ion, which upon loss of a water molecule is expected to give the parent cation H-1+. The hydrogen atoms on the oxonium O atom are involved in strong hydrogen bonds to chlorosulfonate anions and probably partially disordered despite the large estimated pK(a) differences between the corresponding acid-base pairs. The experimentally determined cation structures are compared with structures computed by DFT methods. Detailed experimental procedures are given. PMID- 15339176 TI - Discovery of (E)-9,10-dehydroepothilones through chemical synthesis: on the emergence of 26-trifluoro-(E)-9,10-dehydro-12,13-desoxyepothilone B as a promising anticancer drug candidate. AB - We provide a full account of the discovery of the (E)-9,10-dehydro derivatives of 12,13-desoxyepothilone B (dEpoB), a new class of antitumor agents with promising in vivo preclinical properties. The compounds, which are to date not available by modification of any of the naturally occurring epothilones, were discovered through total chemical synthesis. We describe how our investigations of ring closing metathesis reactions in epothilone settings led to the first and second generation syntheses of (E)-9,10-dehydro-12,13-desoxyepothilone congener 6. With further modifications, the synthesis was applied to reach a 26-trifluoro derivative compound (see compound 7). To conduct such studies and in anticipation of future development needs, the total synthesis which led to the initial discovery of compound 7 was simplified significantly. The total synthesis methodology used to reach compound 7 was then applied to reach more readily formulated compounds, bearing hydroxy and amino functionality on the 21-position (see compounds 45, 62, and 63). Following extensive in vitro evaluations of these new congeners, compound 7 was nominated for in vivo evaluations in xenograft models. The data provided herein demonstrate a promising therapeutic efficacy, activity against large tumors, nonrelapseability, and oral activity. These results have identified compound 7 as a particularly promising compound for clinical development. The excellent, totally synthetic, route to 7 makes such a program quite feasible. PMID- 15339177 TI - Metal-cation-mediated hydrolysis of phosphonoformate diesters: chemoselectivity and catalysis(1). AB - Hydrolyses in D(2)O (pD 1.7-3.1) of dimethyl (7), methyl phenyl (8), phenyl methyl (9), and diphenyl (10) phosphonoformate diesters are substantially accelerated by Ce(IV), Th(IV), Zr(IV), and Hf(IV) cations. Chemoselectivity is observed, whereby Zr(IV) and Hf(IV) principally direct P-OR hydrolysis, whereas Th(IV) and Ce(IV) mainly direct C-OR hydrolysis. Leaving group efficiency (OMe vs OPh) modulates the chemoselectivity. The metal cations also mediate further hydrolytic reactions of the initially produced phosphonoformate monoesters 14, 15, 18, and 20. The origins of the P-OR/C-OR selectivity are discussed in terms of the metal cation hydroxo species likely to be present in solution and the kinetics of the reactions. PMID- 15339178 TI - Investigation on the drying induced phase transformation of mesoporous silica; a comprehensive understanding toward mesophase determination. AB - Phase transformation of mesoporous silica during the drying process is investigated. As-synthesized hexagonal p6mm obtained under the conditions used in this study is transformed to cubic Ia3d as drying proceeds, even at room temperature. Prolonged synthesis results in the formation of a well-ordered hexagonal mesophase, with almost no phase transformation. Drying at a higher temperature promotes the phase transformation of not only hexagonal to cubic, but also cubic to lamellar mesophases. Release of water is detected during drying, which is followed by the phase transformation of the mesophases. The phase transformations observed here proceed against the direction estimated on the basis of the state-of-the-art understanding. Here, considering the degree of silicate condensation and the amount of residual water in the as-synthesized mesoporous silica, a comprehensive explanation of mesophase determination is proposed including thermodynamic and kinetic aspects to account for the results observed here and those in the literature. PMID- 15339179 TI - Supported chiral Mo-based complexes as efficient catalysts for enantioselective olefin metathesis. AB - Syntheses and catalytic activities of seven new polymer-supported chiral Mo-based complexes are disclosed. Four of the complexes are polystyrene-based, and three involve polynorbornene supports. Studies concerning the ability of the polymer bound chiral complexes to promote an assortment of asymmetric ring-closing (ARCM) and ring-opening (AROM) metathesis reactions are detailed. In many instances, levels of reactivity and enantioselectivity are competitive with those of the analogous homogeneous catalysts. The positive effect of lower cross-linking within the polymer backbone on reaction efficiency and asymmetric induction is detailed. The optically enriched products obtained through the use of the supported complexes, after simple filtration and removal of the supported Mo catalysts, contain significantly lower levels of metal impurities as compared to products synthesized with the corresponding homogeneous catalysts. PMID- 15339180 TI - Combined computational and experimental studies of the mechanism and scope of the retro-Nazarov reaction. AB - Density functional theory calculations (B3LYP/6-31+G) demonstrate that conjugating and electron-donating substituents at carbons three and four of a cyclopentenyl oxyallylic cation should have a rate-accelerating effect on the retro-Nazarov reactions of these species. The retro-Nazarov reaction of these intermediates is predicted to exhibit significant torquoselectivity when carbon three is substituted with a methoxy and a methyl group. Experimental studies show that oxyallylic cations can undergo effective retro-Nazarov reactions when two alkyl and one aryl/vinyl groups are on carbons three and four. An equal number of alkyl substituents or a single aryl substituent is not effective in promoting the reaction. Interestingly, a single alkoxy substituent at carbon three is sufficient for the retro-Nazarov reaction to occur. The methodology developed was used in a total synthesis of the natural product turmerone. PMID- 15339181 TI - Label-free colorimetric detection of specific sequences in genomic DNA amplified by the polymerase chain reaction. AB - We document the surprising result that single-stranded DNA adsorbs on negatively charged gold nanoparticles (Au-nps) with a rate that depends on sequence length and temperature. After ss-DNA adsorbs on Au-nps, we find that the particles are stabilized against salt-induced aggregation. These observations can be rationalized on the basis of electrostatics and form the basis for a colorimetric assay to identify specific sequences and single nucleotide polymorphisms on polymerase chain reaction (PCR)-amplified DNA. The assay is label-free, requires no covalent modification of the DNA or Au-np surfaces, and takes on the sensitivity of PCR. Most important, binding of target and probe takes place in solution where hybridization occurs in less than 1 min. As an example, we test PCR-amplified genomic DNA from clinical samples for single nucleotide polymorphisms (SNPs) associated with a fatal arrhythmia known as long QT syndrome. PMID- 15339182 TI - Driving forces for self-organized coadsorption: C6H6/2O and C6H6/2CO on Ni[111]. AB - The self-organized (2log3 x 2log3) coadsorbed phases of C(6)H(6) with O and with CO are investigated within first-principles density functional theory. The main driving force for formation of the C(6)H(6)/2O phase is found to be the reduction of O adatom repulsive interactions, while for the C(6)H(6)/2CO phase it is the interspecies attractive interactions and benzene-benzene repulsive interactions which are most important. PMID- 15339183 TI - Solid-state Ru-99 NMR spectroscopy: a useful tool for characterizing prototypal diamagnetic ruthenium compounds. AB - The feasibility of (99)Ru NMR spectroscopy as a tool to characterize solid compounds is demonstrated. Results of the first solid-state (99)Ru NMR investigation of diamagnetic compounds are presented for Ru(NH(3))(6)Cl(2), K(4)Ru(CN)(6). xH(2)O (x = 0, 3), LaKRu(CN)(6), and Ru(3)(CO)(12). The sensitivity of the ruthenium magnetic shielding tensor to subtle changes in the local structure about the ruthenium nucleus is highlighted by comparing the (99)Ru isotropic chemical shift of Ru(NH(3))(6)Cl(2) in aqueous solutions and in the solid state. The narrow isotropic (99)Ru NMR peak observed for solid Ru(NH(3))(6)Cl(2) indicates that this compound is an ideal secondary reference sample for solid-state (99)Ru NMR studies. The isotropic (99)Ru chemical shift, (99)Ru nuclear quadrupolar coupling constant, C(Q), and quadrupolar asymmetry parameter of K(4)Ru(CN)(6). xH(2)O (x = 0, 3) are shown to be sensitive to x. For Ru(3)(CO)(12), the magnetic shielding tensors of each of the three nonequivalent Ru nuclei have spans of 1300-1400 ppm, and the (99)Ru C(Q) values are also similar, 1.36-1.85 MHz, and are surprisingly small given that (99)Ru has a moderate nuclear quadrupole moment. Information about the relative orientation of the Ru magnetic shielding and electric field gradient tensors has been determined for Ru(3)(CO)(12) from experimental (99)Ru NMR spectra as well as quantum chemical calculations. PMID- 15339184 TI - Vibrational spectroscopy and density functional theory of transition-metal ion benzene and dibenzene complexes in the gas phase. AB - Metal-benzene complexes of the form M(benzene)(n) (M = Ti, V, Fe, Co, Ni) are produced in the gas-phase environment of a molecular beam by laser vaporization in a pulsed nozzle cluster source. These complexes are photoionized with an ArF excimer laser, producing the corresponding cations. The respective mono- and dibenzene complex ions are isolated in an ion-trap mass spectrometer and studied with infrared resonance enhanced multiple-photon dissociation (IR-REMPD) spectroscopy using a tunable free electron laser. Photodissociation of all complexes occurs by the elimination of intact neutral benzene molecules, and this process is enhanced on resonances in the vibrational spectrum, making it possible to measure vibrational spectroscopy for size-selected complexes. Vibrational bands in the 600-1700 cm(-1) region are characteristic of the benzene molecular moiety with systematic shifts caused by the metal bonding. The spectra in this solvent-free environment exhibit periodic trends in band shifts and intensities relative to the free benzene molecule that varies with the metal. Density functional theory calculations are employed to investigate the structures, energetics, and vibrational frequencies of these complexes. The comparison between experiment and theory provides fascinating new insight into the bonding in these prototypical organometallic complexes. PMID- 15339185 TI - Theoretical study of the stereoselective additions of chiral alcohols to ketenes. AB - In 1989, Larsen et al. at Merck discovered that the addition of chiral alcohols to ketenes provided enantiomerically enriched aryl propionic acids, compounds valued for their therapeutic antiinflammatory properties. The high 1,4-asymmetric induction observed (>99:1 dr in the addition, and up to 99% ee after hydrolysis to the acid) is rare. A quantitative model based on B3LYP density functional theory calculations accounts for the stereoselectivity in the addition of (S) methyl lactate, (S)-3-methyl-2-butanol, and (S)-pantolactone to methylphenylketene. The conformational processes of the intermediates can impact the stereoselectivity of the process, and either the addition step, or the protonation of the enolate intermediate, may be stereoselectivity determining. PMID- 15339186 TI - Formation of an interlocked quadruplex dimer by d(GGGT). AB - A tetranucleotide sequence d(GGGT) has been shown to self-assemble into an interlocking quadruplex dimer. UV-melting studies indicated the existence of two species that each showed distinct quadruplex melting transitions, a low-T(m) species, Q(l), and a high-T(m) species, Q(h). Conditions were controlled to favor the formation of either Q(l) or Q(h). Q(l) and Q(h) each showed circular dichroism spectra characteristic of parallel quadruplexes. Negative ion nano electrospray ionization mass spectrometry confirmed that Q(l) was a tetrameric complex, d(GGGT)(4), and Q(h) was an octameric complex, d(GGGT)(8). High resolution (1)H NMR spectroscopy evidenced that d(GGGT)(4) was a C(4)-symmetric parallel tetramolecular quadruplex. The (1)H NMR spectrum of d(GGGT)(8) was consistent with a structure formed by the dimerization of a parallel, "slipped" tetramolecular quadruplex that has its diagonal strands staggered by one base. This "slippage" results in two guanine bases at the 5' end of the quadruplex being presented diagonally that are not involved in tetrads. Two such "slipped" quadruplexes dimerize via these free G-bases at the 5' ends by forming an extra G tetrad. Each "slipped" quadruplex contributes two guanine bases to this extra G tetrad. The formation of a novel GTGT tetrad is also observed at both the 3' ends of the interlocked quadruplex dimer. PMID- 15339187 TI - Binding of distamycin A to UV-damaged DNA. AB - We have found that distamycin A can bind to DNA duplexes containing the (6-4) photoproduct, one of the major UV lesions in DNA, despite the changes, caused by photoproduct formation, in both the chemical structure of the base moiety and the local tertiary structure of the helix. A 20-mer duplex containing the target site, AATT.AATT, was designed, and then one of the TT sequences was changed to the (6-4) photoproduct. Distamycin binding to the photoproduct-containing duplex was detected by CD spectroscopy, whereas specific binding did not occur when the TT site was changed to a cyclobutane pyrimidine dimer, another type of UV lesion. Distamycin binding was analyzed in detail using 14-mer duplexes. Curve fitting of the CD titration data and induced CD difference spectra revealed that the binding stoichiometry changed from 1:1 to 2:1 with photoproduct formation. Melting curves of the drug-DNA complexes also supported this stoichiometry. PMID- 15339188 TI - A single hydroxyl group governs ligand site selectivity in human ileal bile acid binding protein. AB - The recognition between proteins and their native ligands is fundamental to biological function. In vivo, human ileal bile acid binding protein (I-BABP) encounters a range of bile salts that vary in the number and position of steroidal hydroxyl groups and the presence and type of side-chain conjugation. Therefore, it is necessary to understand how chemical variability in the ligand affects the energetic and structural aspects of its recognition. Here we report studies of the binding site selectivity of I-BABP for glycocholic (GCA) and glycochenodeoxycholic (GCDA) acids using isotope-enriched bile salts along with two-dimensional heteronuclear NMR methods. When I-BABP is presented with either GCA or GCDA alone, the ligands bind to both sites. However, when presented with an equimolar mixture of the two bile salts, GCDA binds exclusively to site 1 and GCA to site 2. This remarkable selectivity is governed by the presence or absence of a single hydroxyl group at the C-12 position of the steroid tetracycle. The basis for this site selectivity appears to be energetic rather then steric. PMID- 15339189 TI - Concerning the mechanism of the ring opening of propylene oxide in the copolymerization of propylene oxide and carbon dioxide to give poly(propylene carbonate). AB - The reactions between (TPP)AlX, where TPP = tetraphenylporphyrin and X = Cl, O(CH(2))(9)CH(3), and O(2)C(CH(2))(6)CH(3), and propylene oxide, PO, have been studied in CDCl(3) and have been shown to give (TPP)AlOCHMeCH(2)X and (TPP)AlOCH(2)CHMeX compounds. The relative rates of ring opening of PO follow the order Cl > OR > O(2)CR, but in the presence of added 4-(dimethylamino)pyridine, DMAP (1 equiv), the order is changed to O(2)CR > OR. From studies of kinetics, the ring opening of PO is shown to be first order in [Al]. Carbon dioxide inserts reversibly into the Al-OR bond to give the compound (TPP)AlO(2)COR, and this reaction is promoted by the addition of DMAP. The coordination of DMAP to (TPP)AlX is favored in the order O(2)C(CH(2))(6)CH(3) > O(2)CO(CH(2))(9)CH(3) >> O(CH(2))(9)CH(3). The microstructure of the poly(propylene carbonate), PPC, formed in the reactions between (TPP)AlCl/DMAP and (R,R-salen)CrCl and rac-PO/S PO/R-PO and CO(2), has been investigated by (13)C [(1)H] NMR spectroscopy. The ring opening of PO is shown to proceed via competitive attack on the methine and methylene carbon atoms, and furthermore attack at the methine carbon occurs with both retention and inversion of stereochemistry. On the basis of these results, the reaction pathway leading to ring opening of PO can be traced to an interchange associative mechanism, wherein coordination of PO to the electrophilic aluminum atom occurs within the vicinity of the Al-X bond (X = Cl, OR, O(2)CR, or O(2)COR). The role of DMAP is two-fold: (i) to labilize the trans Al-X bond toward heterolytic behavior, and (ii) to promote the insertion of CO(2) into the Al-OR bond. PMID- 15339190 TI - Hydrolytic reactions of guanosyl-(3',3')-uridine and guanosyl-(3',3')-(2',5'-di-O methyluridine). AB - Hydrolytic reactions of guanosyl-(3',3')-uridine and guanosyl-(3',3')-(2',5'-di-O methyluridine) have been followed by RP HPLC over a wide pH range at 363.2 K in order to elucidate the role of the 2'-hydroxyl group as a hydrogen-bond donor upon departure of the 3'-uridine moiety. Under neutral and basic conditions, guanosyl-(3',3')-uridine undergoes hydroxide ion-catalyzed cleavage (first order in [OH(-)]) of the P-O3' bonds, giving uridine and guanosine 2',3'-cyclic monophosphates, which are subsequently hydrolyzed to a mixture of 2'- and 3' monophosphates. This bond rupture is 23 times as fast as the corresponding cleavage of the P-O3' bond of guanosyl-(3',3')-(2',5'-di-O-methyluridine) to yield 2',5'-O-dimethyluridine and guanosine 2',3'-cyclic phosphate. Under acidic conditions, where the reactivity differences are smaller, depurination and isomerization compete with the cleavage. The effect of Zn(2+) on the cleavage of the P-O3' bonds of guanosyl-(3',3')-uridine is modest: about 6-fold acceleration was observed at [Zn(2+)] = 5 mmol L(-)(1) and pH 5.6. With guanosyl-(3',3') (2',5'-di-O-methyluridine) the rate-acceleration effect is greater: a 37-fold acceleration was observed. The mechanisms of the partial reactions, in particular the effects of the 2'-hydroxyl group on the departure of the 3'-linked nucleoside, are discussed. PMID- 15339191 TI - Exploring CH-activation pathways in bifunctional zirconocene/borane systems. AB - The dimethylsilanediyl-bridged ansa-zirconocene dichloride 1, that contains a pendent allyl substituent at a Cp-ring, adds HB(C(6)F(5))(2) to the vinyl group to yield the bifunctional zirconocene/borane complex 2. Substituted benzimidazoles were added to the strongly electrophilic borane moiety as protective groups, which allowed subsequent chloride versus -CH(2)SiMe(3) exchange at zirconium to take place by treatment with the respective alkyllithium reagent. Alternatively, the introduction of active sigma-ligands at zirconium is carried out first, followed by the hydroboration reaction. This route was followed for the synthesis of the diphenyl-ansa-zirconocene/borane complex 12. Complex 12 reacts slowly in solution by intramolecular electrophilic attack of the borane at its adjacent Cp-ring, followed by deprotonation using a [Zr]-Ph group to yield the zwitterionic complex 14 featuring a borata-tetrahydroindenyl moiety as part of the ansa-metallocene framework. Complex 14 was characterized by X-ray diffraction. It adds PMe(3) at zirconium to yield 15. Thermolysis of 12 with excess PMe(3) leads to the formation of the (aryne)zirconocene complex 18, which is stabilized by PMe(3) coordination to zirconium and PMe(3) addition to boron. N-Methylbenzimidazole adds to the -B(C(6)F(5))(2) unit of 12 to give the 1:1 adduct 19. Thermolysis of 19 at 80 degrees C in benzene solution in the presence of one additional equivalent of N-methylbenzimidazole results in deprotonation of the substrate to yield the sigma-N-methylbenzimidazolyl zirconium complex 20 (as a mixture of two diastereoisomers). An additional N methylbenzimidazole ligand is bonded to the B(C(6)F(5))(2) unit in this product. PMID- 15339192 TI - Catalytic intermolecular Pauson-Khand-type reaction: strong directing effect of pyridylsilyl and pyrimidylsilyl groups and isolation of Ru complexes relevant to catalytic reaction. AB - Some circumstantial evidence for the directing effect of the 2-pyridylsilyl group in the Ru-catalyzed intermolecular Pauson-Khand-type reaction (PKR) of alkenyl(2 pyridyl)silane, alkyne, and carbon monoxide has been provided. Most importantly, we have succeeded in isolating several monometallic Ru complexes relevant to the catalytic reaction: Ru(vinylsilane)(CO)(3) complexes and ruthenacyclopentene. While the stoichimetric reaction of the Ru(vinylsilane)(CO)(3) complex with an alkyne led to the formation of the corresponding cyclopentenone (PKR product) at 100 degrees C, the ruthenacyclopentene intermediate was quantitatively produced at 50 degrees C. This complex was also converted to a cyclopentenone upon heating at 100 degrees C. Moreover, it was also found that the Ru(vinylsilane)(CO)(3) complex and ruthenacyclopentene serve as catalysts in intermolecular PKR. PMID- 15339193 TI - On the reaction mechanism of adduct formation in LOV domains of the plant blue light receptor phototropin. AB - The blue-light sensitive photoreceptor, phototropin, is a flavoprotein which regulates the phototropism response of higher plants. The photoinduced triplet state and the photoreactivity of the flavin-mononucleotide (FMN) cofactor in two LOV domains of Avena sativa, Adiantum capillus-veneris, and Chlamydomonas reinhardtii phototropin have been studied by time-resolved electron paramagnetic resonance (EPR) and UV-vis spectroscopy at low temperatures (T < or = 80 K). Differences in the electronic structure of the FMN as reflected by altered zero field splitting parameters of the triplet state could be correlated with changes in the amino acid composition of the binding pocket in wild-type LOV1 and LOV2 as well as in mutant LOV domains. Even at cryogenic temperatures, time-resolved EPR experiments indicate photoreactivity of the wild-type LOV domains, which was further characterized by UV-vis spectroscopy. Wild-type LOV1 and LOV2 were found to form an adduct between the FMN cofactor and the functional cysteine with a yield of 22% and 68%, respectively. The absorption maximum of the low-temperature photoproduct of wild-type LOV2 is red-shifted by about 15 nm as compared with the FMN C(4a)-cysteinyl adduct formed at room temperature. In light of these observations, we discuss a radical-pair reaction mechanism for the primary photoreaction in LOV domains. PMID- 15339194 TI - "Through-space" nuclear spin-spin J(PP) coupling in tetraphosphine ferrocenyl derivatives: a (31)P NMR and X-ray structure correlation study for coordination complexes. AB - Herein, we report on (31)P(31)P solution-phase "through-space" nuclear spin-spin coupling constants (J(PP)) from a novel family of organometallic tetraphosphine nickel and palladium complexes. These J(PP) constants were accurately determined through NMR iterative simulation based on the second-order spectra obtained for the compounds. The corresponding solid-state X-ray structures of the complexes were determined, and the "through-space" P.P distances are reported. Due to the blocked conformation of the species in solution, a qualitative and semiquantitative experimental correlation is obtained, which links the geometric parameters and the intensity of the corresponding P.P coupling constant. The lone pair overlap theory developed for (19)F(19)F and (15)N(19)F "through-space" couplings in organic compounds [J. Am. Chem. Soc. 1973, 95, 7747-7752; 2000, 122, 4108-4116] appears to be a reliable foundation on which to account for our results. Based on the reported observations, the lone-pair overlap model is extended to "through-space" (31)P(31)P coupling, and the model is broadened to encompass metal orbital contributions for coordination complexes. Some of the predictions and consequences of the proposed theory are discussed. PMID- 15339195 TI - Magnetic field effects on cytochrome c-mediated bioelectrocatalytic transformations. AB - Constant magnetic fields affect many biological transformations, but we lack mechanistic understanding of the processes. The magnetohydrodynamic effect may account for the enhancement of bioelectrocatalytic transformations at interfaces. This is exemplified by the bioelectrocatalyzed cytochrome c-mediated reduction of oxygen and oxidation of lactate in the presence of cytochrome oxidase and lactate dehydrogenase, respectively. We observe significant magnetic field effects on the rates of bioelectrochemical transformations (ca. 3-fold increase) at the functionalized interfaces at field strengths, B, up to 1 T. We show that the limiting current is proportional to the B(1/3)C*(4/3), where C is the concentration of electroactive species. The results may have important implications on the understanding of the magnetic field effects on natural biocatalytic processes at membranes and on the enhancement of biotransformations in biotechnology. PMID- 15339196 TI - Nuclear magnetic shielding and quadrupole coupling tensors in liquid water: a combined molecular dynamics simulation and quantum chemical study. AB - Nuclear magnetic resonance (NMR) shielding tensors for the oxygen and hydrogen nuclei, as well as nuclear quadrupole coupling tensors for the oxygen and deuterium nuclei of water in the liquid and gaseous state, are calculated using Hartree-Fock and density functional theory methods, for snapshots sampled from Car-Parrinello molecular dynamics trajectories. Clusters representing local liquid structures and instantaneous configurations of a single molecule representing low-density gas are fed into a quantum chemical program for the calculation of the NMR tensors. The average isotropic and anisotropic tensorial properties of 400 samples in both states, averaged using a common Eckart coordinate frame, are calculated from the data. We report results for the gas-to liquid chemical shifts of (17)O and (1)H nuclei, as well as the corresponding change in the nuclear quadrupole couplings of (17)O and (2)H. Full thermally averaged shielding and quadrupole coupling tensors are reported for the gaseous and liquid-state water, for the first time in the case of liquid. Electron correlation effects, the difference of classical vs quantum mechanical rovibrational averaging, and different methods of averaging anisotropic properties are discussed. PMID- 15339197 TI - Direct immobilization of native yeast iso-1 cytochrome C on bare gold: fast electron relay to redox enzymes and zeptomole protein-film voltammetry. AB - Cyclic voltammetry shows that yeast iso-1-cytochrome c (YCC), chemisorbed on a bare gold electrode via Cys102, exhibits fast, reversible interfacial electron transfer (k(0) = 1.8 x 10(3) s(-1)) and retains its native functionality. Vectorially immobilized YCC relays electrons to yeast cytochrome c peroxidase, and to both cytochrome cd(1) nitrite reductase (NIR) and nitric oxide reductase from Paracoccus denitrificans, thereby revealing the mechanistic properties of these enzymes. On a microelectrode, we measured nitrite turnover by approximately 80 zmol (49 000 molecules) of NIR, coadsorbed on 0.65 amol (390 000 molecules) of YCC. PMID- 15339199 TI - Role of influenza vaccine for healthy children in the US. AB - Influenza infection is associated with significant morbidity and mortality in adults, but the highest attack rates for influenza regularly occur in children, particularly those in preschool and elementary school. The consequences of influenza in this younger population - increased rate of hospitalization in those younger than 2 years of age and serious associated morbidity - have been underestimated. Children are also the critical link for spreading influenza in the community. Recent data suggest that mass influenza vaccination of healthy children would not only protect recipients, but also may reduce the burden of influenza throughout the community. During the past 3 decades, efforts to control influenza have focused on the use of an injectable trivalent inactivated vaccine (TIV) in high-risk persons. The vaccine is 'safe' and effective, but its acceptance and uptake by patients and healthcare providers have been modest at best. A new intranasal, live-attenuated, trivalent cold-adapted influenza virus vaccine (CAIV-T) [FluMist] is 'safe', well tolerated, immunogenic, and efficacious in preventing influenza illness in healthy children. Compared with TIV, CAIV-T is easier to administer and should be more readily acceptable, particularly for mass immunization campaigns. CAIV-T also induces a broader immune response and has demonstrated protection against at least three different variant influenza strains. This vaccine is particularly well suited for routine immunization of children and thus offers the potential for greatly improved control of influenza. However, the acquisition cost per single dose of FluMist for the 2003-4 season ( approximate, equals 46 US dollars) significantly hampered its uptake both by practitioners and by managed care organizations, even despite a later approximate, equals 25 US dollars rebate offer. For the 2004-5 season, CAIV-T is likely to be only modestly more expensive (average wholesale price: 16.50 US dollars for non-returnable doses, 23 US dollars for returnable doses) than TIV. The practitioner must consider the benefits of FluMist compared with its likely higher vaccine cost and the issues of reimbursement among multiple insurers. PMID- 15339200 TI - Central precocious puberty: current treatment options. AB - Central precocious puberty (CPP) is characterized by early pubertal changes, acceleration of growth velocity, and rapid bone maturation that often result in reduced adult height. An onset of pubertal signs before the age of 8 years in girls and 9 years in boys should always be evaluated. A combination of clinical signs, bone age, pelvic echography in girls, and hormonal data are required to diagnose CPP and make a judgment concerning progression and prognosis. Not all children with apparently true CPP require medical intervention. The main reasons for treatment are to prevent compromised adult height and to avoid psychosocial or behavioral problems. The need for treatment for auxologic reasons is based on estimation of predicted adult height, with the finding of a reduced height potential, which may require a follow-up. Indication for treatment on the basis of psychologic and behavioral anomalies has to be determined on an individual basis. The main short-term aims of therapy are to stop the progression of secondary sex characteristics and menses (in girls) and to treat the underlying cause, when known. Long-term goals are to increase final adult height and to promote psychosocial well-being. Once it has been decided that treatment is appropriate, it should be initiated immediately with depot gonadotropin-releasing hormone (GnRH) agonists. The effective suppression of pituitary gonadal function is achieved with these compounds in practically all CPP patients. Long-term data are now available from 2 decades of GnRH agonist treatment for patients with CPP. Treatment preserves height potential in the majority of patients (especially in younger patients) and improves the final adult height of children with rapidly progressing CPP, with a complete recovery of the hypothalamic-pituitary-gonadal axis after treatment. GnRH agonist treatment using depot preparations is useful and has a good safety profile, with minimal adverse effects and no severe long term consequences. Although further data are need, there may be a role in the future for combining somatropin (growth hormone) and GnRH agonist treatment for some patients with significantly impaired growth velocity. The introduction of GnRH antagonists is likely to improve the treatment options for CPP. PMID- 15339203 TI - Chemical and structural changes in blood undergoing laser photocoagulation. AB - The treatment of cutaneous vascular lesions (port wine stains etc.) using lasers has been guided by theories based on the "cold" or room-temperature optical properties of the hemoglobin target chromophore. We have recently presented evidence showing that under the influence of laser irradiation, the optical properties of blood in vitro are time and temperature dependent. Such complications are not currently subsumed into the in vivo theory. Here, we study the time-domain optical properties of blood undergoing photocoagulation in vitro using two newly developed time-resolved techniques. We also study the asymptotic effect of laser photocoagulation on the chemical and structural properties of the components of the blood matrix. We present evidence showing that the photocoagulation process involves significant changes in the optical absorption and scattering properties of blood, coupled with photothermally induced chemical and structural changes. We demonstrate the first use of a laser to deliberately generate magnetic resonance imaging contrast in vitro. We show that this technique offers significant potential advantages to in vivo intravenous chemical contrast agent injection. PMID- 15339204 TI - Metronomic photodynamic therapy as a new paradigm for photodynamic therapy: rationale and preclinical evaluation of technical feasibility for treating malignant brain tumors. AB - The concept of metronomic photodynamic therapy (mPDT) is presented, in which both the photosensitizer and light are delivered continuously at low rates for extended periods of time to increase selective tumor cell kill through apoptosis. The focus of the present preclinical study is on mPDT treatment of malignant brain tumors, in which selectivity tumor cell killing versus damage to normal brain is critical. Previous studies have shown that low-dose PDT using 5 aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) can induce apoptosis in tumor cells without causing necrosis in either tumor or normal brain tissue or apoptosis in the latter. On the basis of the levels of apoptosis achieved and model calculations of brain tumor growth rates, metronomic delivery or multiple PDT treatments, such as hyperfractionation, are likely required to produce enough tumor cell kill to be an effective therapy. In vitro studies confirm that ALA mPDT induces a higher incidence of apoptotic (terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate, sodium salt nick-end labeling positive) cells as compared with an acute, high-dose regimen (ALA-aPDT). In vivo, mPDT poses two substantial technical challenges: extended delivery of ALA and implantation of devices for extended light delivery while allowing unencumbered movement. In rat models, ALA administration via the drinking water has been accomplished at very high doses (up to 10 times therapeutic dose) for up to 10 days, and ex vivo spectrofluorimetry of tumor (9L gliosarcoma) and normal brain demonstrates a 3-4 fold increase in the tumor-to-brain ratio of PpIX concentration, without evidence of toxicity. After mPDT treatment, histological staining reveals extensive apoptosis within the tumor periphery and surrounding microinvading colonies that is not evident in normal brain or tumor before treatment. Prototype light sources and delivery devices were found to be practical, either using a laser diode or light-emitting diode (LED) coupled to an implanted optical fiber in the rat model or a directly implanted LED using a rabbit model. The combined delivery of both drug and light during an extended period, without compromising survival of the animals, is demonstrated. Preliminary evidence of selective apoptosis of tumor under these conditions is presented. PMID- 15339201 TI - Allergic rhinitis in children : diagnosis and management strategies. AB - The incidence of allergic rhinitis has been increasing for the last few decades, in keeping with the rising incidence of atopy worldwide. Allergic rhinitis has a prevalence of up to 40% in children, although it frequently goes unrecognized and untreated. This can have enormous negative consequences, particularly in children, since it is associated with numerous complications and comorbidities that have a significant health impact on quality of life. In fact, allergic rhinitis is considered to be a risk factor for asthma. There are numerous signs of allergic rhinitis, particularly in children, that can alert an observant clinician to its presence. Children with severe allergic rhinitis often have facial manifestations of itching and obstructed breathing, including a gaping mouth, chapped lips, evidence of sleep deprivation, a long face, dental malloclusions, and the allergic shiner, allergic salute, or allergic crease. The medical history is extremely important as it can reveal information regarding a family history of atopy and the progression of atopy in the child. It is also important to identify the specific triggers of allergic rhinitis, because one of the keys to successful management is the avoidance of triggers. A tripartite treatment strategy that embraces environmental control, immunotherapy, and pharmacologic treatment is the most comprehensive approach. Immunotherapy has come to be viewed as potentially prophylactic, capable of altering the course of allergic rhinitis. The most recent guidelines for the management of allergic rhinitis issued by the WHO recommend a tiered approach that integrates diagnosis and treatment, in which allergic rhinitis is subclassified both by frequency, as either intermittent or persistent, and by severity, as either mild or moderate to severe. Oral or topical antihistamines and intranasal corticosteroids are the mainstay of pharmacologic therapy for allergic rhinitis, depending upon its severity, and several agents have been approved for use in children aged 5 years old and younger. PMID- 15339202 TI - Meningococcal vaccines. AB - Meningococcal disease is one of the most feared and serious infections in the young and its prevention by vaccination is an important goal. The high degree of antigenic variability of the organism makes the meningococcus a challenging target for vaccine prevention. Meningococcal polysaccharide vaccines against serogroup A and C are efficacious and have been widely used, often in combination with serogroup Y and W135 components. Their relative lack of immunogenicity in young children and infants can be overcome by conjugation to a protein carrier. The effectiveness of serogroup C glycoconjugate vaccines in children of all ages has been demonstrated and they have now been introduced into routine vaccination schedules. Conjugate vaccines against other serogroups, including A, Y, and W135 will soon be available and it is hoped they may emulate this success. Prevention of serogroup B disease has proven more elusive. Several serogroup B vaccines based on outer membrane vesicles have been shown to be immunogenic and reasonably effective in adults and older children, but the protection offered by them is chiefly strain-specific. Multivalent recombinant PorA vaccines have been developed to broaden the protective effect, but no efficacy data are available as yet. Intensive efforts have been directed at other outer membrane protein vaccine candidates and lipopolysaccharide, and some of these have been shown to offer protection in experimental animal models. Nonpathogenic Neisseriae spp. such as Neisseria lactamica are also possible vaccine candidates. Previously unknown proteins have been identified from in silico analysis of the meningococcal genome and their vaccine potential explored. However, none of these has yet been presented as the 'universal' protective antigen and work in this field continues to be held back by our limited knowledge concerning the mechanisms of natural protection against serogroup B meningococci. PMID- 15339205 TI - Rose Bengal-sensitized photooxidation of the dipeptides L-tryptophyl-L phenylalanine, L-tryptophyl-L-tyrosine and L-tryptophyl-L-tryptophan: kinetics, mechanism and photoproducts. AB - The Rose Bengal-sensitized photooxidations of the dipeptides l-tryptophyl-l phenylalanine (Trp-Phe), l-tryptophyl-l-tyrosine (Trp-Tyr) and l-tryptophyl-l tryptophan (Trp-Trp) have been studied in pH 7 water solution using static photolysis and time-resolved methods. Kinetic results indicate that the tryptophan (Trp) moiety interacts with singlet molecular oxygen (O(2)((1)Delta(g))) both through chemical reaction and through physical quenching, and that the photooxidations can be compared with those of equimolecular mixtures of the corresponding free amino acids, with minimum, if any, influence of the peptide bond on the chemical reaction. This is not a common behavior in other di- and polypeptides of photooxidizable amino acids. The ratio between chemical (k(r)) and overall (k(t)) rate constants for the interaction O(2)((1)Delta(g))-dipeptide indicates that Trp-Phe and Trp-Trp are good candidates to suffer photodynamic action, with k(r)/k(t) values of 0.72 and 0.60, respectively (0.65 for free Trp). In the case of Trp-Tyr, a lower k(r)/k(t) value (0.18) has been found, likely as a result of the high component of physical deactivation of O(2)((1)Delta(g)) by the tyrosine moiety. The analysis of the photooxidation products shows that the main target for O(2)((1)Delta(g)) attack is the Trp group and suggests a much lower accumulation of kynurenine-type products, as compared with free Trp. This is possibly because of the occurrence of another accepted alternative pathway of oxidation that gives rise to 3a oxidized hydrogenated pyrrolo[2,3-b]indoles. PMID- 15339206 TI - Spatial control of reactive oxygen species formation in fibroblasts using two photon excitation. AB - Two-photon excitation (2PE) provides a means of generating reactive oxygen species (ROS) in cells and tissues with a high degree of spatial specificity. In cultured monolayers of human fibroblasts and fibroblast-derived cells treated with the commonly used probe of ROS formation, 5-(and 6)-chloromethyl-2',7' dichlorodihydrofluorescein diacetate, acetyl ester (CM-H(2)DCFDA), cells irradiated through a microscope objective with 150 fs near-infrared laser pulses became highly fluorescent, reflecting intracellular ROS formation. The fluorescence intensity inside cells increased quadratically with the average power of radiation for pulsed excitation and was unchanged for continuous wave irradiation with the same average power. Single fibroblasts embedded within dermal equivalents were also targeted in this manner and formed ROS, whereas neighboring unirradiated cells were spared. These results demonstrate that ROS can be generated intracellularly in skin cells using 2PE of the metabolic or oxidative products of CM-H(2)DCFDA and that formation of ROS can be localized in both cell monolayers and in a tissue equivalent. This technique should be useful in understanding the response of whole tissues such as skin to local generation of ROS and may have applications in photodynamic therapy. PMID- 15339207 TI - Virus particles monitored by fluorescence spectroscopy: a potential detection assay for macromolecular assembly. AB - Native fluorescence spectroscopy was used for in situ investigations of two lipid containing bacteriophages from the cystovirus family as well as their Pseudomonad host cells. Both the viruses phi6 and phi12 and their bacterial host proteins contain the amino acid tryptophan (trp), which is the predominant fluorophore in UV. Within proteins, trp's structural environment differs, and the differences are reflected in their spectroscopic signatures. It was observed that the peak of the trp emission from both viruses was at 330 nm, a significantly shorter wavelength than trp in either the Pseudomonad host cells or the amino acid's chemical form. This allowed us to monitor the viral attachment process and subsequent lytic release of progeny virus particles by measurement of the trp emission spectra during the infection process. This work demonstrates that fluorescence may offer a novel tool to detect viruses and monitor viral infection of cells and may be part of a biodefense application. PMID- 15339208 TI - Identification of genes responsive to UV-A radiation in human lens epithelial cells using complementary DNA microarrays. AB - UV-A radiation produces cataract in animals, enhances photoaging of the lens and skin and increases the phototoxicity of drugs. However, the nature of genes that are activated or repressed after cellular exposure to UV-A radiation remains enigmatic. Because lens epithelial cells exposed to UV-A radiation undergo apoptosis 4 h after exposure to the stress, we sought to establish the change in gene expression in cells by UV-A radiation using gene expression profiling using complementary DNA microarrays containing about 12 000 genes. We identified 78 genes abnormally expressed in UV-A-irradiated cells (showing >2.5-fold change at P < 0.05). These genes are implicated in various biological processes, including signal transduction and nucleic acid binding, and genes encoding enzymes. A majority of the genes were downregulated. Our analysis revealed that the expression of genes for the transcription factors ATF-3 and Pilot increased four fold, whereas the gene for the apoptosis regulator NAPOR-1 decreased five-fold. These changes were confirmed by real-time quantitative reverse transcriptase polymerase chain reaction. The calpain large polypeptide 3 (CANP3) gene also increased nine-fold after UV-A radiation. In addition, peroxisomal biogenesis factor 7, glucocorticoid receptor-alpha and tumor-associated calcium signal transducer genes decreased three- to eight-fold. Western blot analysis further confirmed the increase in protein expression of ATF-3 and CANP3 and decreased expression of glucocorticoid receptor-alpha in the irradiated cells. Surprisingly, most of these genes had not been previously shown to be modulated by UV-A radiation. Our results show that human lens epithelial cells respond to a single dose of UV-A radiation by enhancing or suppressing functionally similar sets of genes, some of which have opposing functions, around the time at which apoptosis occurs. These studies support the intriguing concept that activation of competing pathways favoring either cell survival or death is a means to coordinate the response of cells to UV-A stress. PMID- 15339209 TI - Dermatological risk of indoor ultraviolet exposure from contemporary lighting sources. AB - Discussions of risks and implications of cutaneous exposure to indoor lighting, including hypothetical contribution to causality of melanoma, have mainly concentrated on ultraviolet (UV) A and B (UVA, UVB) spectral emissions from fluorescent bulbs. Only studies of quartz halogen lamps have suggested that users might sustain UVC-induced injury. Examination of light sources in the home and school of a child with xeroderma pigmentosum revealed that several different types emitted surprising levels of UV. Our purpose was to assess the extent of UV emissions from a variety of commonly used light sources to identify potential dermatological risks. UV and visible spectral emissions of commercially obtained lamps of several types were measured using a calibrated spectral radiometer traceable to the National Institute of Standards and Technology. Indoor light sources including fluorescent, quartz halogen and even tungsten filament incandescent lamps provided UVA, UVB and sometimes UVC emissions. Intensities of some emissions were of similar magnitude to those in sunlight. Chronic exposure to indoor lighting may deliver unexpected cumulative UV exposure to the skin and eyes. Patients with UV-exacerbated dermatoses should be cautioned about potential adverse reactions from indoor lighting. PMID- 15339210 TI - Energy dissipation processes of singlet-excited 1-hydroxyfluorenone and its hydrogen-bonded complex with N-methylimidazole. AB - Effects of solvent, pH and hydrogen bonding with N-methylimidazole (MIm) on the photophysical properties of 1-hydroxyfluorenone (1HOF) have been studied. Fluorescence lifetime, fluorescence quantum yield and triplet yield measurements demonstrated that intersystem crossing was the dominant process in apolar media and its rate constant significantly diminished with increasing solvent polarity. The acceleration of internal conversion in alcohols paralleled the strength of intermolecular hydrogen bonding. The faster energy dissipation from the singlet excited state in cyclohexane was attributed to intramolecular hydrogen bonding. The pK(a) of 1HOF decreased from 10.06 to 5.0 on light absorption, and H(3)O(+) quenched the singlet-excited molecules in a practically diffusion-controlled reaction. On addition of MIm in toluene, dual fluorescence was observed, which was attributed to reversible formation of excited hydrogen-bonded ion pair. Rate constants for the various deactivation pathways were derived from the combined analysis of the steady-state and the time-resolved fluorescence results. PMID- 15339211 TI - The spatial distribution of optical properties in the ultraviolet and visible in an aquatic ecosystem. AB - In aquatic ecosystems, the UV and visible radiation environment is strongly influenced by variation in the chemical and physical parameters of the ecosystems. In shallow lakes, highly heterogeneous water characteristics produce a wide variety of optical environments. Such ecosystems require analysis approaches that consider a potential variability. In this study, 77 stations were used to characterize the optical properties of a shallow lake (open water surface 54 km(2)). The vertical attenuation of solar radiation at 305, 313, 320 and 340 nm and at photosynthetically active radiation was measured during the seasonal cycle. Dissolved organic matter (DOM), turbidity, fluorescence, pH, temperature, conductance and dissolved oxygen were simultaneously measured. The spatial variation of the extinction spectra of the dissolved fraction at each sampling station was also measured and analyzed between 270 and 400 nm. The spatial heterogeneity of the lake was examined by determining the distributions of the attenuation coefficients and biooptical parameters at high spatial resolution and describing the distributions in a series of maps. The methodology permitted a quantitative description of the interaction between solar radiation and aquatic ecosystems as well as a spatial classification of the dominating processes within the lake. This included the determination of the role played by DOM loading and changing chemical properties within the lake optical environment. PMID- 15339212 TI - Suppression of different phases of systemic contact hypersensitivity by urocanic acid oxidation products. AB - On exposure to UV-B, the epidermal component trans-urocanic acid (UCA) is not only photoisomerized into cis-UCA but will also, at least in part, be photooxidized into UCA oxidation products (UOPs). We hypothesized that UOPs can mimic UV-induced systemic immunosuppression comparable to the suppressive properties already established for cis-UCA. A crude mixture of UOPs showed a significant suppression of the sensitization phase of the systemic contact hypersensitivity (CHS) response to picryl chloride (PCl). Three of the UOPs were selected for this study: imidazole-4-carboxylic acid (ImCOOH), imidazole-4 carboxaldehyde (ImCHO) and imidazole-4-acetic acid (ImAc). Effects on the sensitization, elicitation and postelicitation phases of CHS to PCl in BALB/c mice were studied and compared with the effects of cis-UCA. ImCHO was equally effective at suppressing the sensitization phase as cis-UCA. The triplet combination of the imidazoles (1:1:1) showed more pronounced suppression than that induced by cis-UCA. The most effective compounds for the suppression of the elicitation phase appeared to be ImAc and cis-UCA. Significant suppression of the postelicitation phase was only obtained with the triplet combination of ImCHO, ImCOOH and ImAc, the combination that appeared to be effective at all three tested phases. Because these three UOPs are present in UV-B-exposed human stratum corneum, these compounds may play a role in UV-B-induced immunosuppression. PMID- 15339213 TI - Characterization of the Fe(III)-binding site in Sepia eumelanin by resonance Raman confocal microspectroscopy. AB - The resonance Raman spectrum of Sepia eumelanin is discussed by analogy to model compounds containing catechol (CAT)-like structural units. These data are then compared with the analogous data on Fe(III)-enriched Sepia eumelanin. In contrast to the natural eumelanin, the Fe(III)-enriched samples exhibit absorption features in the visible and near-IR spectral regions, which are attributed to ligand-to-metal charge-transfer (LMCT) bands. Resonance Raman spectra collected by exciting these LMCT bands reveal bands at 580 and 1470 cm(-1); the intensity of these features increases with increasing Fe(III) content. The 580 and 1470 cm( 1) bands are assigned to Fe-OR stretching and ring deformation modes, respectively. These data further substantiate that the Fe(III)-melanin-binding site in melanin is composed of CAT-like structural units. PMID- 15339214 TI - Flexibility in proteins: tuning the sensitivity to O2 diffusion by varying the lifetime of a phosphorescent sensor in horseradish peroxidase. AB - The heme in horseradish peroxidase (HRP) was replaced by phosphorescent Pt mesoporphyrin IX (PtMP), which acted as a phosphorescent marker of oxygen quenching and allowed comparison with another probe, Pd-mesoporphyrin IX (Khajehpour et al. (2003) Proteins 53, 656-666). Benzohydroxamic acid (BHA), a competitive inhibitor of the enzyme, was also used to monitor its effects on phosphorescence quenching. With the addition of BHA, in the presence of oxygen, the phosphorescence intensity of the protein increased. In contrast, the addition of BHA, in the absence of oxygen, reduced the phosphorescence intensity of the protein. K(d) = 18 microM when BHA binds to PtMP-HRP. The effect of BHA can be explained by two factors: (1) BHA reduces the accessibility of O(2) to the protein interior and (2) BHA itself quenches the phosphorescence. Consistent with this, the oxygen quenching of the phosphorescence of PtMP-HRP gave a quenching constant of k(q) = 234 mm Hg(-1) s(-1) in the absence of BHA and k(q) = 28.7 mm Hg(-1) s(-1) in the presence of BHA. The quenching rate of BHA is 4000 s(-1). The relative quantum yield of the phosphorescence of the Pt derivative is about six times that of the Pd derivative, whereas the phosphorescence lifetime is approximately eight times shorter. The high quantum yield and suitable lifetime make Pt-porphyrins appropriate as sensors of O(2) diffusion and flexibility in heme proteins. PMID- 15339215 TI - Riboflavin and UV-light based pathogen reduction: extent and consequence of DNA damage at the molecular level. AB - We are developing a technology based on the combined application of riboflavin (RB) and light for inactivating pathogens in blood products while retaining the biological functions of the treated cells and proteins. Virus and bacteria reduction measured by tissue culture infectivity or colony formation with UV light alone and in combination with RB yield equivalent results. The effects of RB as a sensitizing agent on DNA in white cells, bacteria and viruses in combination with UV light exposure have been evaluated. UV-mediated DNA degradation in Jurkat T cells and leukocytes in plasma as measured by the FlowTACS assay was significantly increased in the presence of RB. Agarose gel electrophoretic analysis of DNA in Escherichia coli and leukocytes in plasma demonstrated enhanced DNA degradation in the presence of RB. UV light in combination with RB prevented the reactivation of lambda phage compared with samples irradiated in the absence of RB. UV-mediated oxidative damage in calf thymus DNA was also enhanced in the presence of RB. These observations clearly demonstrate that the presence of RB and UV light selectively enhances damage to the guanine bases in DNA. These data also suggest that the type and extent of damage to DNA for virus in the presence of RB and light make it less likely to be repaired by normal repair pathways available in host cells. PMID- 15339216 TI - Fluorescence lifetime spectroscopy of glioblastoma multiforme. AB - Fluorescence spectroscopy of the endogenous emission of brain tumors has been researched as a potentially important method for the intraoperative localization of brain tumor margins. We investigated the use of time-resolved, laser-induced fluorescence spectroscopy for demarcation of primary brain tumors by studying the time-resolved spectra of gliomas. The fluorescence of human brain samples (glioblastoma multiforme, cortex and white matter: six patients, 23 sites) was induced ex vivo with a pulsed nitrogen laser (337 nm, 3 ns). The time-resolved spectra were detected in a 360-550 nm wavelength range using a fast digitizer and gated detection. Parameters derived from both the spectral- (intensities from narrow spectral bands) and the time domain (average lifetime) measured at 390 and 460 nm were used for tissue characterization. We determined that high-grade gliomas are characterized by fluorescence lifetimes that varied with the emission wavelength (>3 ns at 390 nm, <1 ns at 460 nm) and their emission is overall longer than that of normal brain tissue. Our study demonstrates that the use of fluorescence lifetime not only improves the specificity of fluorescence measurements but also allows a more robust evaluation of data collected from brain tissue. Combined information from both the spectral- and the time domain can enhance the ability of fluorescence-based techniques to diagnose and detect brain tumor margins intraoperatively. PMID- 15339217 TI - A multicomponent model of chromophoric dissolved organic matter photobleaching. AB - Light absorption by chromophoric dissolved organic matter (CDOM) plays a number of roles in natural waters, including both control of the underwater light field and the initiation of many photochemical reactions. A multicomponent analysis was used to describe the effects of UV and visible radiation on the optical absorption spectra of two natural water samples, a Suwannee River fulvic acid standard (SRFA) and a Delaware Bay water sample. This analysis used a constrained minimization technique to fit independent spectral components to the observed bleaching behavior of the water samples under monochromatic irradiation. Spectra derived from these fits were used to predict the bleaching behavior of both samples under polychromatic irradiation (lambda > 320 nm). This approach reproduces the kinetics and spectral behavior of polychromatic photobleaching very well at times <48 h, but underpredicts the bleaching at longer time periods. PMID- 15339218 TI - Effects of UV-B irradiation on a marine microecosystem. AB - Purpose of this work was to study the effect of UV irradiation on a microecosystem consisting of several interacting species. The system chosen was of a hypersaline type, where all the species present live at high salt concentration; it comprises different bacteria; a producer, the photosynthetic green alga Dunaliella salina; and a consumer, the ciliated protozoan Fabrea salina, which form a complete food chain. We were able to establish the initial conditions that give rise to a self-sustaining microecosystem, stable for at least 3 weeks. We then determined the effect of UV irradiation on this microecosystem under laboratory-controlled conditions, in particular by measuring the critical UV exposure for the two main components of the microecosystem (algae and protozoa) under UV-B irradiances comparable to those of solar irradiation. In our experiments, we varied irradiance, total dose and spectral composition of the actinic light. The critical doses at irradiances of the order of 56 kJ/m(2) (typical average daily irradiance in a sunny summer day in Pisa), measured for each main component of the microecosystem (algae and ciliates), turned out to be around 70 kJ/m(2) for ciliates and 50 kJ/m(2) for D. salina. By exposing microecosystems to daily UV-B irradiances of the order of 8 kJ/m(2) (typical average daily irradiance in a sunny winter day in Pisa), we found no effect at total doses of the order of the critical doses at high irradiances, showing that the reciprocity law does not hold. We have also measured a preliminary spectral sensitive curve of the UV effects, which shows an exponential decay with wavelength. PMID- 15339220 TI - Effects of supplementary ultraviolet-B irradiance on maize yield and qualities: a field experiment. AB - Stratospheric ozone depletion has caused an increase in the amount of ultraviolet B (UV-B) radiation reaching the earth's surface. Numerous investigations have demonstrated that the effect of UV-B enhancements on plants includes reduction in grain yield, alteration in species competition, susceptibility to disease and changes in plant structure and pigmentation. Many experiments examining UV-B radiation effects on plants have been conducted in growth chambers or greenhouses. It has been questioned whether the effect of UV-B radiation on plants can be extrapolated to field responses from indoor studies because of the unnaturally high ratios of UV-B/ultraviolet-A radiation (320-400 nm) and UV B/photosynthetically active radiation (PAR) in many indoor studies. Field studies on UV-B radiation effect on plants have been recommended to use the UV and PAR irradiance provided by natural light. This study reports the growth and yield responses of a maize crop exposed to enhanced UV-B radiation and the UV-B effects on maize seed qualities under field conditions. Enhanced UV-B radiation caused a significant reduction in the dry matter accumulation and the maize yield in turn was affected. With increased UV-B radiation the flavonoid accumulation in maize leaves increased and the contents of chlorophyll a, b and (a + b) of maize leaves were reduced. The levels of protein, sugar and starch of maize seed decreased with enhanced UV-B radiation, whereas the level of lysine increased with enhanced UV-B radiation. PMID- 15339221 TI - Light modulates electric phenomena in hornet cuticle. AB - In the cuticle of live social hornets, such as Vespa orientalis (Hymenoptera, Vespinae), endogenous electric effects are encountered, i.e. voltages of 100-200 mV under illumination and currents amounting to several microamperes on its subjection to darkness-clearly a process of charging and discharging. Of the various wavelengths of sunlight, UV was found to be the most contributory to the active cuticular voltage generation. Throughout the warm season of the year--the active period in colonies of social hornets and wasps--colony members exit from the dark nest during the daytime and fly to the field under the hot sun for various foraging purposes, ultimately returning to the nest. Thus, each hornet, be it queen, worker or drone, probably undergoes daily cyclical process of electric charge and discharge in the exterior part of their integument, cuticle, which lasts up to 30-40 min. Such photoelectric phenomenon was detected in both live, ether-anaesthetized hornets and dead hornets, albeit in the latter the electric values recorded were lower. The present study addresses the possible impact of the phenomenon on vespan daily life and also compares it with a parallel occurrence in electric fish. PMID- 15339222 TI - Photophysical properties of coumarin-30 dye in aprotic and protic solvents of varying polarities. AB - Experimental results on various photophysical properties of coumarin-30 (C30) dye, namely, Stokes' shift (Deltanu), fluorescence quantum yield (Phi(f)), fluorescence lifetime (tau(f)), radiative rate constant (k(f)) and nonradiative rate constant (k(nr)), as obtained using absorption and fluorescence measurements have been reported. Though in most of the solvents the properties of C30 show more or less linear correlation with the solvent polarity function, Deltaf = [(epsilon - 1)/(2epsilon + 1) - (n(2) - 1)/(2n(2) + 1)], they show unusual deviations in nonpolar solvents at one end and in high-polarity protic solvents at the other end. From the solvent polarity and temperature effect on the photophysical properties of the dye, following inferences have been drawn: (1) in nonpolar solvents, the dye exists in a nonpolar structure, where its 7-NEt(2) substituent adopts a pyramidal configuration and the amino lone pair is out of resonance with the benzopyrone pi cloud; (2) in medium to higher polarity solvents, the dye exists in a polar intramolecular charge transfer structure, where the 7-NEt(2) group and the 1,2-benzopyrone moiety are in the same plane and the amino lone pair is in resonance with the benzopyrone pi cloud; (3) in protic solvents, the dye-solvent intermolecular hydrogen bonding influences the photophysical properties of the dye; and (4) in high-polarity protic solvents, the excited C30 undergoes a new activation-controlled nonradiative deexcitation process because of the involvement of a twisted intramolecular charge transfer (TICT) state. Contrary to most other TICT molecules, the activation barrier for this deexcitation process in C30 is observed to increase with solvent polarity. A rational for this unusual behavior has been given on the basis of the solvent polarity-dependent stabilization and crossing of relevant electronic states and the relative propensity of interconversion among these states. PMID- 15339223 TI - Tryptophan fluorescence in the Bacillus subtilis phototropin-related protein YtvA as a marker of interdomain interaction. AB - The Bacillus subtilis protein YtvA, related to plant phototropins (phot), binds flavin mononucleotide (FMN) within the N-terminal light, oxygen and voltage (LOV) domain. The blue light-triggered photocycle of YtvA and phot involves the reversible formation of a covalent photoadduct between FMN and a cysteine (cys) residue. YtvA contains a single tryptophan, W103, localized on the LOV domain and conserved in all phot-LOV domains. In this study, we show that the fluorescence parameters of W103 in YtvA-LOV are markedly different from those observed in the full-length YtvA. The fluorescence quantum yields are ca 0.03 and 0.08, respectively. In YtvA-LOV, the maximum is redshifted (ca 345 vs 335 nm) and the average fluorescence lifetime shorter (2.7 vs 4.7 ns). These data indicate that W103 is located in a site of tight contact between the two domains of YtvA. In the FMN-cys adduct, selective excitation of W103 at 295 nm results in minimal changes of the fluorescence parameters with respect to the dark state. On 280 nm excitation, however, there is a detectable decrease in the fluorescence emitted from tyrosines, with concomitant increase in W103 fluorescence. This effect is reversible in the dark and might arise from a light-regulated energy transfer process from a yet unidentified tyrosine to W103. PMID- 15339224 TI - Structural heterogeneity of cryotrapped intermediates in the bacterial blue light photoreceptor, photoactive yellow protein. AB - We investigate by X-ray crystallographic techniques the cryotrapped states that accumulate on controlled illumination of the blue light photoreceptor, photoactive yellow protein (PYP), at 110 K in both the wild-type species and its E46Q mutant. These states are related to those that occur during the chromophore isomerization process in the PYP photocycle at room temperature. The structures present in such states were determined at high resolution, 0.95-1.05A. In both wild type and mutant PYP, the cryotrapped state is not composed of a single, quasitransition state structure but rather of a heterogeneous mixture of three species in addition to the ground state structure. We identify and refine these three photoactivated species under the assumption that the structural changes are limited to simple isomerization events of the chromophore that otherwise retains chemical bonding similar to that in the ground state. The refined chromophore models are essentially identical in the wild type and the E46Q mutant, which implies that the early stages of their photocycle mechanisms are the same. PMID- 15339225 TI - The fluorescence properties of hypocrellin B and its amino-substituted derivative: photoinduced intramolecular proton transfer and photoinduced intramolecular electron transfer. AB - We report on the emission spectra and emission quantum yields of a newly synthesized hypocrellin dye, 2-demethoxy-2,3-ethylenediamino hypocrellin B (EDAHB), and its parent HB in different solvents of varying polarity. Our results demonstrate that EDAHB is one of the few dyes that exhibit highly solvent polarity-dependent fluorescence in the useful region (680-730 nm). Therefore, it offers some applications in the biomedical field as a fluorescent probe molecule. The solvatochromic effect of EDAHB is proposed to be due to a distinct change in the dipole moment of the dye on excitation. A photoinduced intramolecular proton transfer and a photoinduced intramolecular electron transfer process are considered relevant for the fluorescence properties of HB and EDAHB, respectively. PMID- 15339226 TI - Expressed sequence tag analysis of the dinoflagellate Lingulodinium polyedrum during dark phase. AB - To collect information on gene expression during the dark period in the luminous dinoflagellate Lingulodinium polyedrum, normalized complementary DNA (cDNA) libraries were constructed from cells collected during the first hour of night phase in a 12:12 h light-dark cycle. A total of 4324 5'-end sequence tags were isolated. The sequences were grouped into 2111 independent expressed sequence tags (EST) from which 433 groups were established by similarity searches of the public nonredundant protein database. Homology analysis of the total sequences indicated that the luminous dinoflagellate is more similar to land plants and animals (vertebrates and invertebrates) than to prokaryotes or algae. We also isolated three bioluminescence-related (luciferase and two luciferin-binding proteins [LBP]) and 37 photosynthesis-related genes. Interestingly, two kinds of LBP genes occur in multiple copies in the genome, in contrast to the single luciferase gene. These cDNA clones and EST sequence data should provide a powerful resource for future genome-wide functional analyses for uncharacterized genes. PMID- 15339228 TI - Velopharyngeal incompetence: the need for speech standards. AB - Speech-language pathologists are often involved in the diagnosis and treatment of velopharyngeal incompetence (VPI). Some speech-language pathologists have extensive educational and clinical experience with VPI; others have limited training and experience. Thus, the quality of speech-language services for people with VPI is heterogenous, and it ranges from poor to excellent. There are, as yet, no specific guidelines for speech-language pathologists providing services to people with VPI. Optimal services require specific guidelines for training and experience. The purpose of this article was to (a) review speech-language pathology standards and qualifications, (b) provide reasons for identifying qualified speech-language pathologists, and (c) identify strategies for reduction of risks involved in the delivery of speech-language services for people with VPI. PMID- 15339229 TI - Effect-size reporting practices in AJSLP and other ASHA journals, 1999-2003. AB - A census of effect-size practices in the past 5 volumes of American Speech Language-Hearing Association journals was accomplished. Inclusion of effect size in quantitative research reports increased from 5 reports with effect size in 1990 to 1994 to 120 reports in 1999 to 2003. Nonetheless, effect size was reported less than 30% of the time when inferential statistics were used, and only half of those reports included an interpretation of effect size. This article presents case exemplars to illustrate the use and value of effect size and includes suggestions for interpreting effect size. Researchers are encouraged to routinely report effect size and to interpret effect size in a way that facilitates the application of research to practice. PMID- 15339230 TI - The voice use reduction program. AB - The purpose of this article is to describe a structured behavior modification approach to the reduction of voice use by clients with voice disorders. The Voice Use Reduction (VUR) Program is conceptualized as part of a comprehensive approach to the treatment of voice. The VUR Program provides guidelines for the classification of voice use situations, the assignment of voice use units to different situations, and the calculation of the maximum number of units per day and per week in a severe, moderate, and low voice use reduction program. Two case examples are described to illustrate the application of the VUR Program. The results of an evaluation of the VUR Program by 10 female students who presented with vocal nodules and applied the program also are included. PMID- 15339231 TI - Self-rating of stuttering severity as a clinical tool. AB - Scaling is a convenient and equipment-free means for speech-language pathologists (SLPs) and clients to evaluate stuttering severity in everyday situations. This study investigated the extent to which the severity ratings of 10 adult stuttering speakers, made immediately after speaking and again from recordings 6 months later, agreed with ratings made by an SLP. For 9 of the 10 speakers, there was good agreement between their initial ratings and those of the SLP. For 8 of the 10 speakers, there was also good agreement between their initial ratings and those made from recordings 6 months later, indicating that the severity ratings made at the time of speaking were reliable. These findings support the use of the 9-point scale as a clinical measurement procedure. PMID- 15339232 TI - Gesture-speech combinations as a transition to multiword utterances. AB - This study was designed to determine if certain types of gesture-speech combinations act as transitional phenomena preceding production of 2-word utterances. Ten normally developing children with a mean age of 15 months at the beginning of the study participated in this research. The children were sampled longitudinally at monthly intervals as they approached the onset of early multiword utterances. Temporally synchronized gesture-speech combinations were analyzed over a 6-month period to describe whether they encoded 1 semantic element (pointing to a car and saying "car") or 2 semantic elements (pointing to a car and saying "big"). These gesture-speech combinations were examined in terms of their onset in relation to early multiword combinations. It was found that there was a significant increase in gesture-speech combinations encoding 2 semantic elements during the 6-month period and that the onset of these combinations preceded or co-occurred with the 1st productions of multiword utterances. This finding, coupled with prior studies on smaller numbers of participants, suggests that gesture-speech combinations encoding 2 elements may be a transitional element between single-word communication and the onset of multiword combinations. PMID- 15339233 TI - Semantic feature analysis treatment for anomia in two fluent aphasia syndromes. AB - The effect of semantic feature analysis (SFA) treatment on confrontation naming and discourse production was examined in 2 persons, 1 with anomic aphasia and 1 with Wernicke's aphasia. Results indicated that confrontation naming of treated nouns improved and generalized to untreated nouns for both participants, who appeared to have different lexical access impairments. Both participants demonstrated improvement in some aspects of discourse production associated with the confrontation naming SFA treatment. However, there was no change in most manifestations of lexical retrieval difficulty during discourse for either participant. These findings support previous work regarding improved and generalized naming associated with SFA treatment and indicate a need to examine effects of improved confrontation naming on more natural speaking situations. PMID- 15339234 TI - Effect of phonemic perception training on the speech production and phonological awareness skills of children with expressive phonological delay. AB - Children with expressive phonological delays often possess poor underlying perceptual knowledge of the sound system and show delayed development of segmental organization of that system. The purpose of this study was to investigate the benefits of a perceptual approach to the treatment of expressive phonological delay. Thirty-four preschoolers with moderate or severe expressive phonological delays received 16 treatment sessions in addition to their regular speech-language therapy. The experimental group received training in phonemic perception, letter recognition, letter-sound association, and onset-rime matching. The control group listened to computerized books. The experimental group showed greater improvements in phonemic perception and articulatory accuracy but not in phonological awareness in comparison with the control group. PMID- 15339235 TI - Evidence-based clinical practice: a Canadian perspective. PMID- 15339237 TI - Genetic defects in GH synthesis and secretion. AB - Growth hormone (GH) is a multifunctional hormone produced in the anterior pituitary that promotes postnatal growth of skeletal and soft tissues. GH secretion and release are complex phenomena depending on several intrinsic and extrinsic factors modulating the release of two hypothalamic hormones, GH releasing hormone and somatostatin. Any genetic or acquired disorder that impairs GH secretion or action causes a pathological phenotype characterized by harmonic short stature with isolated GH deficiency (IGHD) or combined pituitary hormone deficiency (CPHD). In this article we report current knowledge about the genetic basis of IGHD and CPHD. PMID- 15339238 TI - New molecular mechanisms of GH resistance. AB - Primary growth hormone (GH) resistance describes growth failure in the presence of normal, or even elevated, GH secretion. In its classic form, the phenotype is identical to that of GH deficiency, and was originally described in association with defects of the GH receptor. With increasing understanding of the GH-insulin like growth factor (IGF) axis, it has become apparent that GH resistance can result from either primary IGF deficiency (IGFD) or IGF resistance. Primary IGFD may be due to: (i) defects of the GH receptor, (ii) defects of post-GH receptor signaling or (iii) primary defects of IGF-I synthesis. IGF resistance may result from: (i) defects of the IGF receptor, (ii) defects of post-IGF receptor signaling, (iii) defects of IGF binding proteins or (iv) defects of the epiphyseal growth plate or of regulatory proteins involved in epiphyseal growth. PMID- 15339239 TI - Evidence for a link between IGF-I and cancer. AB - Cancer risk is determined by a combination of environmental factors and genetic predisposition. Recent evidence suggests that dietary and related factors such as physical activity and body size may influence cancer risk through their effects on the serum concentration of IGF-I and its binding proteins. The growth hormone (GH)/IGF-I axis is involved in both human development as well as the maintenance of normal function and homeostasis in most cells of the body. In addition to their classical role as endocrine hormones, its members regulate a wide range of biological functions such as cell proliferation, differentiation and apoptosis through paracrine and autocrine mechanisms. During cancer development this complex network regulating tissue homeostasis breaks down, with inappropriate expression of the GH/IGF-I axis making an important contribution. The increased understanding of the molecular mechanisms and signalling pathways regulated by the GH/IGF-I axis has started to provide significant insights into the aetiology, prevention and therapy for a number of common cancers. PMID- 15339240 TI - The GH/IGF-I axis and longevity. AB - Several converging lines of evidence obtained over the last years in a wide variety of experimental model organisms suggest that the ageing process is regulated by genes that encode proteins from the somatotroph axis: longevity genes like daf-2, which were identified using mutant Caenorhabditis elegans strains, turned out to be orthologues of the mammalian genes encoding insulin like signalling cascade proteins. Transgenic flies with mutations in the corresponding insect genes showed a similar pattern of increased lifespan. Finally, mice with spontaneous mutations leading to pituitary hormone deficiency significantly outlived controls. While these and other genetic models suggest that the downregulation of the somatotroph axis can slow the ageing process, other results from studies using pharmacological administration of growth hormone suggest that such stimulating treatment can restore some of the phenotypic traits associated with youth. To better understand the role of the insulin-like receptors in mammalian lifespan regulation and ageing, we explored the phenotype of heterozygous IGF-I receptor (IGF1R) knockout mice. Compared with control littermates these mutants live longer without any obvious impairment of their health and physiology, except a reduced glucose tolerance that we observed in males. These IGF1R(+/-) mutants were also more resistant to oxidative stress in vivo, and we identified a possible molecular pathway linking underphosphorylation of IGF-I receptors to the lack of activation of p66Shc, a protein capable of increasing resistance to oxidative stress through regulation of a set of downstream genes. These and other results suggest that in mammals too, lifespan can be increased by continuous, long-term downregulation of IGF signalling. Since growth hormone administration normally stimulates IGF production in tissues, the question arises whether the beneficial effects of GH, as reported by others, could be IGF independent. This hypothesis can be addressed, for example, by adequately combining existing transgenic mouse models. PMID- 15339241 TI - The psychological burden of short stature: evidence against. AB - Short stature, per se, is clearly not a disease, but is commonly perceived to be associated with social and psychological disadvantage. The assumption, widely held by pediatricians that short children are likely to be significantly affected by their stature, has been founded largely on older, poorly designed clinic-based studies and laboratory investigations of beliefs about the association between stature and individual characteristics. In contrast, data from more recent and better designed clinic- and community-based studies show that, in terms of psychosocial functioning, individuals with short stature are largely indistinguishable from their peers, whether in childhood, adolescence or adulthood. Parents and children alike should be reassured by these findings. In the absence of clear pathology, physical or psychological, GH therapy for the short but otherwise normal child raises ethical concerns about so-called 'cosmetic endocrinology'. PMID- 15339242 TI - Why do some children of short stature develop psychologically well while others have problems? AB - The present paper addresses the question: why do some children of short stature develop psychologically well while others have problems? Based on the work of Wallander and Varni, a model is presented to illustrate risk as well as resistance factors that are important for children of short stature. It is suggested that important risk factors for the psychological adjustment of children of short stature are the child's satisfaction with its height and the aetiology of the short stature. Another possible risk factor is the tendency for people in the child's environment to treat the child as if he or she were younger than is actually the case. The most important risk factor, however, seems to be the psychosocial stress related to being teased or bullied due to the short stature. Important resistance factors for children of short stature might be the child's temperament, familial support and coping strategies. It is concluded that an important aim for future research is, in a multi-disciplinary setting, to empirically test models of risk and resistance factors that are relevant for children of short stature. PMID- 15339243 TI - Benefit of postponing normal puberty for improving final height. AB - Experiments of nature and clinical observations have provided indications that postponing puberty may increase final height in short children. In children with central precocious puberty, a GnRH analog (GnRHa) alone is efficacious in increasing final height, but in other conditions a combination of growth hormone (GH) and GnRHa is needed. In GH-deficient children with early onset of puberty and poor height prediction, the combination of GH and GnRHa increases final height by 1.0-1.3 s.d. In children with idiopathic short stature and persistent short stature after intrauterine growth retardation, the combination also appears to be beneficial. Potential side effects include weight gain, a negative effect on bone mineralization, and psychosocial consequences. More data on long-term safety have to be collected before the combination of GH and GnRHa in children with idiopathic short stature should be considered for clinical use outside clinical trials. PMID- 15339244 TI - Bone development during GH and GnRH analog treatment. AB - Estrogens, GH and IGFs are essential in the development and growth of the skeleton and for the maintenance of bone mass and density. Treatment of precocious puberty with GnRH analogs (GnRHa), by reducing sex steroid levels, leads to a situation of hypoestrogenism that may theoretically have a detrimental effect on bone mass during pubertal development. A reduction in bone mineral density (BMD) during GnRHa treatment has been demonstrated, but GnRHa treatment in patients with central precocious puberty (CPP) does not seem to impair the achievement of normal peak bone mass (PBM) at final height. However, calcium supplementation is effective in improving bone densitometric levels and may promote better PBM achievement. In children and adolescents with GH deficiency (GHD), BMD assessed by dual-energy X-ray absorptiometry (DEXA) and bone turnover are significantly reduced, but they are stimulated by GH treatment. GH treatment leads to improved bone density, function of the dose and duration of treatment, and patients may require prolonged GH treatment beyond the time of growth to improve PBM. After the discontinuation of GH therapy, the more active population had higher bone mineral content (BMC) levels than patients with low physical activity. In our experience, the therapeutic association of GH and calcium also represents a valuable tool in pursuing a proper BMC in GHD patients. We concluded that nonhormonal factors, such as physical activity and nutritional factors, are important in determining bone metabolism and bone mass. PMID- 15339245 TI - Lessons from the national cooperative growth study. AB - OBJECTIVE: To review the National Cooperative Growth Study (NCGS), a national post-marketing surveillance program for children treated with biosynthetic growth hormone (GH) products from Genentech. METHODS: Representative data are presented to summarize the extensive experience of the NCGS. This study is a multicenter, observational surveillance registry begun in 1985 in coordination with the release of the first recombinant DNA biosynthetic GH. RESULTS: After almost 20 years, data from over 47 000 patients representing 165 000 patient years have been collected. There are over 12 000 active subjects (estimated to be approximately 75% of all current patients treated with a Genentech GH product) at 435 centers, providing extensive efficacy and safety data. The GrowTrak program is a secure, user-friendly database which encourages a high level of participation by the pediatric endocrine community in the USA. Efficacy has been shown in a variety of clinical diagnoses, including isolated (IGHD) and organic growth hormone deficiency (OGHD), idiopathic short stature (ISS), and Turner syndrome. Safety monitoring of this large population has provided reassuring evidence that leukemia (de novo or relapse), extracranial nonleukemic neoplasms and central nervous system (CNS) tumor recurrence are not associated with GH therapy. A small risk of intracranial hypertension and slipped capital femoral epiphyses has been suggested, especially in children with renal disease or Turner syndrome. NCGS substudies have also provided important insights into actual clinical practice. For example, screening for renal, cardiac, or auditory comorbidities in Turner syndrome is often done less than is recommended by national guidelines (NCGS 9). Furthermore, almost 65% of children referred for short stature may be lost to follow-up before an evaluation is completed (NCGS 8). CONCLUSIONS: The NCGS has proven to be a valuable method of monitoring the safety and efficacy of biosynthetic GH. The study has attracted wide physician participation due to the data collection software used and the extensive sharing of the analyses of that data with the providers. PMID- 15339246 TI - Transitional care of GH deficiency: when to stop GH therapy. AB - While the benefits of growth hormone (GH) therapy in adult hypopituitary patients with GH deficiency (GHD) are established, the role of continued GH therapy after final height in adolescent GH-deficient patients remains unclear. Preliminary data suggest that cessation of GH on completion of linear growth may be associated with impairment of somatic development and adverse changes in body composition. For the present time, the decision whether to continue GH treatment in adolescent patients with GHD is best made on an individual basis. For such patients, continuity of care is crucial. Children and adults with GHD are usually managed by physicians in separate departments, who may focus on different aspects of treatment and care. Close collaboration between paediatric and adult physicians is essential to ensure smooth transition and to minimize the drop-out rate from follow-up. Given the previous period of treatment during childhood, paediatric physicians should be best placed to discuss the potential benefits of continuing GH therapy and instigate retesting of GH secretion. Many children with isolated idiopathic GHD will produce normal GH responses if retested at adult height. Patients with multiple pituitary hormone deficits are more likely to have ongoing GHD, as are patients who have received CNS irradiation. Quality of life does not appear to be decreased in adolescents with GHD who stop treatment, so achievement of satisfactory bone mass is a major determinant of the decision whether to continue therapy. PMID- 15339247 TI - Ghrelin: update on a novel hormonal system. AB - Ghrelin is a peptide hormone secreted into the circulation from the stomach but also synthesised in a number of tissues, suggesting both endocrine and paracrine effects. These include: stimulation of GH, prolactin and ACTH secretion; an increase in appetite; a diabetogenic effect on carbohydrate metabolism; positive inotropic effects on heart; vasodilatation; and effects on cell proliferation. The possibility of chronic manipulation of the ghrelin system on body weight, growth and appetite remains an exciting new field of exploration. PMID- 15339248 TI - Novel analogs of ghrelin: physiological and clinical implications. AB - Ghrelin, the 28 amino acid peptide recently identified as the natural ligand for the growth hormone (GH) secretagogue (GHS) receptor, has multiple activities in addition to stimulation of GH secretion, including stimulation of feeding and weight gain. To utilize these actions for potential therapeutic benefit, we have produced analogs of human ghrelin with enhanced metabolic stability, affinity for the GHS receptor, and efficacy in stimulating weight gain. We have also discovered an analog of ghrelin, BIM-28163, that is an antagonist at the GHS receptor and that fully inhibits GHS receptor activation induced by native ghrelin. In vivo, BIM-28163 does not increase GH secretion but fully blocks ghrelin-induced GH secretion. In contrast, BIM-28163 acts as a full agonist with regard to the ghrelin actions of stimulating weight gain and food intake. These results suggest that a receptor other than the GHS receptor mediates the actions of ghrelin on feeding and weight gain. This concept is strengthened by our observation that at certain hypothalamic sites, BIM-28163 acts as an antagonist of ghrelin-induced neuronal activation, while at other sites, both ghrelin and BIM-28163 induce neuronal activation via the same receptor. Collectively, these results indicate the existence of a novel ghrelin receptor that may regulate the feeding activity of ghrelin. Using BIM-28163 as a tool to define the endogenous role of ghrelin in normal GH secretion, we have demonstrated that antagonism of the GHS receptor in normal rats does not impair the pulsatility of GH secretion but lowers the pulse amplitude and mean GH level. These results demonstrate that endogenous ghrelin acts to amplify the basic pattern of GH secretion established by the interplay of hypothalamic GH-releasing hormone and somatostatin. These studies demonstrate the feasibility of creating ghrelin analogs that are selective for specific activities, as well as their utility in dissecting the role of ghrelin in both normal physiology and specific pathologies. PMID- 15339249 TI - Cystic fibrosis -- therapeutic challenge in cystic fibrosis children. AB - Cystic fibrosis (CF) is the most common autosomal recessive disease with fatal outcome in Caucasians with a frequency of 1 in 2500 life births. It is caused by mutations in a single gene on the long arm of chromosome 7 encoding a protein called the cystic fibrosis transmembrane regulator (CFTR). The defect in CFTR leads to pathological changes in all organs with mucus-secretory glands, e.g. airways, pancreas, gut, biliary tract, vas deferens and sweat glands. Despite impressive advances in understanding the molecular basis of the disease, life expectancy is still limited in CF and chronic infection of the lung resulting in fibrosis and bronchiectasis followed by respiratory insufficiency is still the main factor in morbidity and the leading cause of death. Poor nutritional status is one of the major problems in the vicious cycle of chronic inflammation and lung destruction and its impact on outcome in lung function has been demonstrated. The possible role of growth hormone treatment in this context will be discussed. PMID- 15339250 TI - GH improves growth and clinical status in children with cystic fibrosis -- a review of published studies. AB - Children with cystic fibrosis (CF) have problems with poor linear growth and inadequate weight gain. Nutritional augmentation has been the mainstay of therapy for improving both weight and height in CF; however, inadequate growth continues to be a problem. Furthermore, protein catabolism has been documented even in non acutely ill adults and children with CF, and could adversely affect longitudinal growth. Human recombinant GH has positive effects on nitrogen balance, and multiple studies have demonstrated improved height and weight in children treated with GH. The purpose of this article is to summarize studies evaluating GH use in children with CF. All published studies of GH use in children with CF have demonstrated significant improvement in height velocity and height Z score. All studies but one, in which subjects were treated only three times per week with GH, have demonstrated improvement in weight as reported by weight velocity and/or weight Z score, and one trial has demonstrated a substantial improvement when GH was used to augment nutritional therapy. Several reports suggest that GH treatment results in improved forced vital capacity, and multiple studies have found improved clinical status as measured by decreased hospitalizations and courses of intravenous antibiotics. Furthermore studies to date also suggest that GH results in improvement in exercise tolerance and bone accumulation. To date significant side effects, including glucose intolerance, have not been reported. Thus mounting evidence suggests that human recombinant GH provides safe and effective therapy in children with CF. PMID- 15339251 TI - The bone mass concept: problems in short stature. AB - Bone densitometry is currently one of the mainstays in the evaluation of systemic bone diseases in adults and is also increasingly used to assess primary or secondary bone disorders in children and adolescents. The purpose of carrying out densitometric studies in such circumstances is to measure the densitometric indicators of bone stability. Following procedures which were established for diagnosing adult osteoporosis, a decrease in densitometric surrogates of bone stability is usually interpreted as indicating increased fracture risk. The most basic densitometric parameter is bone mineral content (BMC), which can be measured with most densitometric techniques. BMC is either defined as the mass of mineral contained in an entire bone or as the mass of mineral per unit bone length. While mineral mass can be expected to be a good surrogate for bone stability, BMC is obviously a size-dependent parameter, since small bones weigh less than big bones. This is a drawback in paediatric use, since many children and adolescents who are examined by densitometry suffer from chronic disorders and are small-for-age. Short children will have a lower BMC than their healthy age-matched peers, even if their (smaller) bones are otherwise completely normal. PMID- 15339252 TI - The heart: an end-organ of GH action. AB - Several experimental and clinical studies have indicate that the heart is an end organ of GH action. Patients with either childhood- or adulthood-onset GH deficiency (GHD) have abnormalities of cardiac structure and function, such as reduced cardiac mass, impaired diastolic filling and reduced left ventricular response at peak exercise. These cardiovascular abnormalities can be reversed, at least partially, after GH replacement therapy. On the other hand, the chronic overproduction of GH and IGF-I in acromegaly leads to the development of a specific cardiomyopathy. Concentric cardiac hypertrophy occurs in more than two thirds of patients at diagnosis and is commonly associated with diastolic dysfunction. In later stages, impaired systolic function ending in heart failure can occur if GH/IGF-I excess is not controlled. Additionally, acromegalic cardiomyopathy is complicated by abnormalities of cardiac rhythm and cardiac valves. Successful control of acromegaly is accompanied by a decrease of the left ventricular mass and improvement of cardiac function. These beneficial effects appear earlier in young patients with short disease duration than in elderly patients. In conclusion, GH and IGF-I play a main role in the regulation of cardiac development and performance. PMID- 15339253 TI - Cytochrome c oxidase subunit IV as a marker of protein kinase Cepsilon function in neonatal cardiac myocytes: implications for cytochrome c oxidase activity. AB - We have previously demonstrated that low concentrations of phorbol esters stimulate the selective translocation of protein kinase C (PKC) alpha and epsilon from the cell soluble to the particulate fraction in NCMs (neonatal rat cardiac myocytes). We therefore determined if the in vitro phosphorylation of substrates in these fractions could be used as assays of PKCalpha or epsilon activation. Intact cell phorbol ester treatment caused a decline in the in vitro (32)P incorporation into several proteins in the cell-soluble fraction. These declines occurred in the presence or absence of in vitro Ca(2+) and probably reflected the exit of PKC isoenzymes from the soluble fraction. In contrast, an approx. 18 kDa protein incorporated (32)P in particulate fractions isolated from 4beta-PMA treated cells in a Ca(2+)-independent manner. Proteomic and immunoprecipitation analyses indicated that the protein is subunit IV of the cytochrome c oxidase complex (COIV). In vitro phosphorylation of COIV was attenuated by PKC pseudosubstrate peptides. Introduction of an PKCepsilon-selective translocation inhibitor [Johnson, Gray, Chen and Mochly-Rosen (1996) J. Biol. Chem. 271, 24962 24966] into NCMs before 4beta-PMA treatments also attenuated the in vitro phosphorylation of COIV. In mitochondrial extracts from 4beta-PMA-treated NCMs, the PKCepsilon isoenzyme coimmunoprecipitated with COIV, and cytochrome c oxidase activity was enhanced 2-fold. The in vitro phosphorylation of COIV reflects a novel approach for monitoring PKCepsilon function in NCMs. Furthermore, PKCepsilon probably interacts with COIV in NCM mitochondria to enhance electron transport chain complex IV activity. PMID- 15339254 TI - Apolipoprotein CI overexpression is not a relevant strategy to block cholesteryl ester transfer protein (CETP) activity in CETP transgenic mice. AB - ApoCI (apolipoprotein CI) is a potent inhibitor of plasma CETP [CE (cholesteryl ester) transfer protein]. The relevance of apoCI overexpression as a method for CETP blockade in vivo was addressed in the present study in CETPTg/apoCITg mice (mice expressing both human CETP and apoCI). Despite a significant reduction in specific CETP activity in CETPTg/apoCITg mice compared with CETPTg mice [transgenic mouse to human CETP; 46.8+/-11.1 versus 101.8+/-25.7 pmol x h( 1).(mug of plasma CETP)(-1) respectively; P<0.05], apoCI overexpression increased both the CETP mass concentration (3-fold increase; P<0.05) and the hepatic CETP mRNA level (4-fold increase, P<0.005), leading to an increase in total plasma CE transfer activity (by 39%, P<0.05). The ratio of apoB-containing lipoprotein to HDL (high-density lipoprotein) CE was 10-fold higher in CETPTg/apoCITg mice than in apoCITg mice (P<0.0005). It is proposed that the increased CETP expression in CETPTg/apoCITg mice is a direct consequence of liver X receptor activation in response to the accumulation of cholesterol-rich apoB-containing lipoproteins. In support of the latter view, hepatic mRNA levels of other liver X receptor responsive genes [ABCG5 (ATP-binding cassette transporter GS) and SREBP-1c (sterol-regulatory-binding protein-1c)] were higher in CETPTg/apoCITg mice compared with CETPTg mice. In conclusion, overexpression of apoCI, while producing a significant inhibitory effect on specific CETP activity, does not represent a suitable method for decreasing total CE transfer activity in CETPTg/apoCITg mice, owing to an hyperlipidaemia-mediated effect on CETP gene expression. PMID- 15339255 TI - Counterpointing the functional role of the forebrain and of the brainstem in the control of the sleep-waking system. AB - This paper reviews the lifetime contributions of the author to the field of sleep wakefulness (S-W), reinterprets results of the early studies, and suggests new conclusions and perspectives. Long-term cats with mesencephalic transection show behavioral/polygraphic rapid eye movement sleep (REMS), including the typical oculo-pupillary behavior, even when the section is performed in kittens prior to S-W maturation. REMS can be induced as a reflex. Typical non-rapid eye movement S (NREMS) is absent and full W/arousal is present only after a precollicular section. The isolated forebrain (IF) rostral to the transection exhibits all features of W/arousal and NREMS [with electroencephalographic (EEG) spindles and delta waves], arousal to olfactory stimuli, and including the appropriate oculo pupillary behaviors. These features also mature normally after neonatal transection. REMS is absent from the IF. After deprivation there is NREMS pressure and rebound in the IF, but the decerebrate cat only shows pressure for REMS. Most IF reactions to pharmacologic agents are within expectations, except for the tolerance/withdrawal effects of long-term morphine use which are absent. In contrast, these effects are supported by the brainstem (i.e. seen in the decerebrate cat). In cats with ablation of the telencephalon, or diencephalic cats, delta waves are absent in the thalamus. EEG thalamic spindle waves are seen triggering S for only 4-5 days after ablation. Therefore, true NREMS is absent in chronic diencephalic cats although pre- and postsomniac behaviors persist. These animals are hyperactive and show a pronounced, permanent insomnia; however, a low dose of barbiturate triggers a dramatic REMS/atypical NREMS rebound. Cats without the thalamus (athalamic cats), initially show a dissociation between behavioral hyperactivity/insomnia and the neocortical EEG, which for 15-20 days exhibits only delta and slower oscillations. Fast, low-voltage W rhythms appear later on, first during REMS, but spindle waves and S postures are absent from the start, such that these cats also display only atypical NREMS. Athalamic cats also show barbiturate-sensitive insomnia. Cats with ablation of the frontal cortices or the caudate nuclei remain permanently hyperactive. They also show a mild, but significant hyposomnia, which is permanent in afrontal cats, but lasts for about a month in acaudates. The polygraphic/behavioral features of their S-W states remain normal. We conclude and propose that: (a) the control of the S-W system is highly complex and distributed, but is organized hierarchically in a well-defined rostro-caudal manner; the rostral-most or highest level (telencephalon), is the most functionally complex/adaptative and regulates the lower levels; the diencephalic/basal forebrain, or middle level, has a pivotal role in inducing switching between S and W and in coordinating the lowest (brainstem) and highest levels; (b) W can occur independently in both the forebrain and brainstem, but true NREMS- and REMS-generating mechanisms exist exclusively in the forebrain and brainstem, respectively; (c) forebrain and brainstem S-W processes can operate independently from each other and are preprogrammed at birth; this helps understanding normal and abnormal polygraphic/behavioral dissociations in humans and normal dissociations/splitting in aquatic mammals; (d) NREMS homeostasis is present in the IF, but only REMS pressure after deprivation persists in the decerebrate cat; (e) the thalamus engages in both NREMS and W; (f) insomnia in diencephalic cats is the result of an imbalance between antagonistic W- and S promoting cellular groups in the ventral brain (normally modulated by the telencephalon); (g) the EEG waves, which are signature for each S-W state, appear to truly drive the concomitant behaviors, e.g. a hypothetical human IF could alternate between behavioral NREMS and W/arousal/awareness; (h) a role for REMS is to keep the individual sleeping at the end of the self-limiting NREMS periods. The need for accelerating research on telencephaling NREMS periods. The need for accelerating research on telencephalic S-W processes and downstream control of the lower S-W system levels is emphasized. PMID- 15339256 TI - Changes in fronto-posterior functional coupling at sleep onset in humans. AB - The aim of this study was to assess the functional coupling between anterior and posterior areas as induced by the sleep onset process. The functional coupling was indexed by an analysis of spectral coherence and directed transfer function (DTF) from electroencephalographic (EEG) data. As it has been reported that more anterior areas first synchronize sleep EEG activity, we hypothesized a fronto posterior direction of the cortical functional coupling during the sleep onset process. Ten normal right-handed male students slept for two-nights (one adaptation, one baseline) in the laboratory, with standard polysomnographic recordings. Spectral coherence and DTF were computed on data recorded by anterior (FzA1) and posterior (PzA1, OzA1) derivations. EEG coherence at the delta/theta band was higher during the presleep period than the sleep onset period, while EEG coherence at the alpha band was higher during the sleep onset period than the presleep period. The DTF findings indicated a prevalence of the occipital-to frontal information flow at delta/theta and alpha bands during the presleep period and a prevalence of the frontal-to-parieto-occipital information flow at all bands during the sleep onset period. The coherent pattern of changes in EEG coherence and in DTF values at sleep onset lends further support to the notion of sleep as a local process, showing that the sleep onset process is subserved by the functional coordination of a cortical fronto-posterior network. In this network, prefrontal areas may play a leading role in the propagation of synchronizing signals conveyed at frequencies spanning delta to beta rhythms. PMID- 15339257 TI - Comparative utility of instruments for monitoring sleepiness-related performance decrements in the operational environment. AB - As both military and commercial operations increasingly become continuous, 24-h per-day enterprises, the likelihood of operator errors or inefficiencies caused by sleep loss and/or circadian desynchrony also increases. Avoidance of such incidents requires the timely application of appropriate interventions--which, in turn, depend on the ability to measure and monitor the performance capacity of individuals in the operational environment. Several factors determine the potential suitability of candidate measures, including their relative sensitivity, reliability, content validity, intrusiveness and cumbersomeness/fieldability. In the present study, the relative sensitivity (defined as the ratio of effect size to 95% confidence interval) of several measures to the effects of sleep loss was compared in a sleep restriction experiment, in which groups were allowed 3, 5, 7, or 9 h time in bed (TIB) across seven consecutive nights. Of the measures compared, the Psychomotor Vigilance Test was among the most sensitive to sleep restriction, was among the most reliable with no evidence of learning over repeated administrations, and possesses characteristics that make it among the most practical for use in the operational environment. PMID- 15339258 TI - Experimentally induced arousals during sleep: a cross-modality matching paradigm. AB - Micro-arousals occur spontaneously or in response to exogenous and endogenous sensory input during sleep. The function of micro-arousals remains unclear, for example, whether it reflects a disturbance or a preparatory response to environmental changes. The goal of this study was to assess arousal responsiveness when two types of sensory stimulations were used: auditory (AD) alone and the addition of a vibrotactile (VT) sensation. Ten normal sleepers participated in three nights of polygraphic recordings. The first night was for habituation and to rule out sleep disorders, and the second to collect baseline sleep data. During the third night, AD and VT + AD stimuli, with three levels of intensities for auditory and vibratory signals, were randomly given to induce arousal responses in sleep stages 2, 3 and 4 and rapid eye movement (REM). The frequency of the arousal responses increased with stimulus intensity for all sleep stages and was lowest in stages 3 and 4. In non-REM (NREM) sleep, combined VT + AD stimulation induced more frequent and more intense arousal responses than AD alone. In REM sleep, more frequent micro-arousals rather than awakenings were triggered by combined stimulations. In stage 2, the response rate of total induced K-complexes did not differ between both types of stimulations while more K-complexes followed by arousals were evoked by the combined VT + AD stimulation than by the AD alone. The induced arousals were associated with an increase in heart rate in all sleep stages. An increase in suprahyoid muscle tone was observed in NREM sleep only, REM being not associated with a rise in muscle tone following experimental stimulation. Most leg and body movements occurred in response to induced awakenings. These results suggest that the cross-modality sensory stimuli triggered more arousal responses in comparison with single modality stimuli. In an attempt to wake a sleeping subject, the addition of a tactile stimulation, such as shaking the shoulder, is an effective strategy that increases the arousal probability. PMID- 15339259 TI - Slow horizontal eye movement at human sleep onset. AB - Slow eye movement (SEM) at human sleep onset has been studied as a function of EEG and behavioral state but has not been subjected to systematic physical measurement. This study counted the frequency of horizontal SEM and quantified its physical properties during hypnagogic EEG stages 3-8 (stage 1 sleep). SEM amplitudes, peak velocities, and durations were digitally measured. As in previous studies SEM occurred throughout stage 1 sleep, coinciding with all tonic states and with all phasic events that classically define stage 1. Within stage 1 the physical properties of SEM varied significantly. In hypnagogic stage 3, where EEG alpha dominates up to half of any 30-s epoch, the typical SEM was moderate in amplitude, slow, and short-lasting, and SEM peak velocity described a weak function of SEM amplitude. In stages 5 and 6 combined--characterized by a low voltage delta-theta EEG, sustained alpha suppression, and no more than one vertex sharp wave (VSW) per 30-s epoch--SEM velocity was maximal, and described a strong function of amplitude. Despite low absolute velocities, stage 5/6 SEM could approximate saccade-like morphologies. In stages 7 and 8 combined, characterized by VSW bursts and incipient spindles or K complexes, SEM was maximal in amplitude and duration but not faster than SEM in stage 5/6. These properties of human SEM are discussed in relation to the anatomy and physiology of SEM in macaque. PMID- 15339260 TI - The 24-h growth hormone rhythm in men: sleep and circadian influences questioned. AB - The 24-h rhythm of growth hormone (GH) is thought to be controlled primarily by sleep processes with a weak circadian component. This concept has been recently questioned in sleep-deprived persons. To test the notion of a high sleep dependency of GH release, we established simultaneous 24-h rhythms of GH and melatonin, a circadian marker, in night workers who form a model for challenging sleep and circadian processes. Ten day-active subjects and 11 night workers were studied during their usual sleep-wake schedule, with sleep from 23:00 to 07:00 hours and 07:00 to 15:00 hours, respectively. Experiments were conducted in sleep rooms under continuous nutrition, bed rest, and dim light. Melatonin and GH were measured every 10 min over 24 h. In day-active subjects, melatonin and GH showed the well-known 24-h profiles, with a major sleep-related GH pulse accounting for 52.8 +/- 3.5% of the 24-h GH production and the onset of the melatonin surge occurring at 21:53 hours +/- 18 min. In night workers, melatonin showed variable circadian adaptation, with the onset of secretion varying between 21:45 and 05:05 hours. The sleep-related GH pulse was lowered, but the reduction was compensated for by the emergence of large individual pulses occurring unpredictably during waking periods, so that the total amount of GH secreted during the 24 h was constant. One cannot predict the degree of GH adaptation from the highly variable melatonin shift. These results argue against the concept that sleep processes exert a predominant influence on GH release whatever the conditions. When sleep and circadian processes are misaligned, the blunting of the sleep-related GH pulse is counteracted, as in sleep-deprived persons, by a compensatory mechanism promoting GH pulses during wakefulness. PMID- 15339261 TI - Corticospinal excitability during laughter: implications for cataplexy and the comparison with REM sleep atonia. AB - Cataplexy is usually seen as rapid eye movement (REM) sleep atonia occurring at an inopportune moment. REM sleep atonia is the result of postsynaptic inhibition, i.e. inhibition of alpha motor neurones. Although this may explain the suppression of H-reflexes during REM sleep, cataplexy and laughter, it is not the only explanation. Presynaptic inhibition, in which afferent impulses are prevented from reaching motor neurones, is an alternative. Testing H-reflexes and magnetic-evoked potentials (MEPs) helps to tell them apart: in postsynaptic inhibition MEPs and H-reflexes change in tandem, while H-reflexes may decrease independent of MEPs with other inhibition modes. We studied motor inhibition during laughter, the strongest trigger for cataplexy. H-reflexes were evoked every 2 s in the soleus muscle in 10 healthy subjects watching comical video fragments. MEPs were evoked when H-reflexes decreased during laughter, and, as a control, when subjects did not laugh. Pairs of MEPs and the immediately preceding H-reflexes were studied. Compared with the control condition, laughter caused mean MEP area to increase by 60% (P=0.006) and mean H-reflex amplitude to decrease by 33% (P=0.008). This pattern proves that postsynaptic inhibition cannot have been the sole influence. The findings do not prove which mechanisms are involved; one possibility is that the decrease in H-reflex amplitude was the result of presynaptic inhibition, and that cortical and/or spinal facilitation accounted for increased MEPs. Regardless, the pattern differs fundamentally from the reported mechanism of REM sleep atonia. Existing scanty data on cataplexy suggest a pattern of H-reflexes and MEPs similar to that during laughter, but this needs further study. PMID- 15339262 TI - Sleep problems in children with autism. AB - Autism is a developmental disability characterized by severe deficits in social interaction and communication, and the presence of repetitive-ritualistic behaviors. Sleep problems are frequently reported by parents of children with autism with prevalence estimates of 44-83% for sleep disorders in this population. To better understand sleep in autism, we surveyed sleep problems in 210 children with autism using a Likert-based questionnaire for parent report. The most frequently reported sleep problems included difficulty in falling asleep, restless sleep, not falling asleep in own bed, and frequent wakenings. Least frequently reported sleep problems were sleep walking, morning headaches, crying during sleep, apnea, and nightmares. When surveys were divided into mental retardation (MR)/not MR categories, no significant differences were identified in frequencies of reported sleep problems except for waking at night which occurred much more frequently in the MR group. There was also no difference in sleep problems related to age of the child other than nocturnal enuresis. An association was noted between certain medical problems and sleep problems. Vision problems, upper respiratory problems, and runny nose were associated with decreased nighttime sleep. Vision problems, poor appetite, and poor growth were associated with increased nighttime waking. Poor appetite and poor growth were associated with decreased willingness to fall asleep. This study confirms a high prevalence of sleep problems reported by parents of children with autism and points to the need for more systematic research as an initial step in developing treatment strategies. PMID- 15339263 TI - Sleep disorders in Taiwanese children with attention deficit/hyperactivity disorder. AB - To assess obstructive sleep apnea syndrome (OSAS) and periodic limb movement disorder (PLMD) in children with attention deficit/hyperactivity disorder (ADHD) compared with a control group. The ADHD was diagnosed based on Diagnostic and Statistical Manual, version IV (DSM-IV) criteria on successively seen elementary school children aged 6-12 years referred to a psychiatric clinic for suspected ADHD. A standardized interview (Kiddie-SADS-E), parents and teacher questionnaires, neuropsychological testing, and nocturnal polysomnography were completed for each child. Eighty-eight children (77 boys) with ADHD and 27 controls were involved in the study. Fifty children with ADHD (56.8%) had an apnea-hypopnea index (AHI) >1 event h(-1) and 17 (19.3%) had an AHI >5 event h( 1). Nine children (10.2%) had a periodic limb movement index (PLMI) >5 events h( 1). There is one child with AHI >1 and none with a PLMI > 5 in the control group. In the test of variables of attention (TOVA), the response time was significantly worse in ADHD with sleep disorders than those without them. The child behavior checklist (CBCL) showed a significant difference between groups in the hyperactivity subscale. The diagnostic criteria for ADHD based on DSM-IV do not differentiate between children with or without sleep disorders. Evaluation of sleep disorders should be considered before starting drug treatment for ADHD. PMID- 15339264 TI - Analysis of postarousal EEG activity during somnambulistic episodes. AB - Early studies found that electroencephalographic (EEG) recordings during somnambulistic episodes were characterized by a combination of alpha, theta, and delta frequencies, without evidence of clear wakefulness. Three postarousal EEG patterns associated with slow-wave sleep (SWS) arousals were recently identified in adults with sleepwalking and sleep terrors. The goal of the present study was to evaluate the distribution of these postarousal EEG patterns in 10 somnambulistic patients (three males, seven females, mean age: 25.1, SD: 4.1) evaluated at baseline and following 38 h of sleep deprivation. A total of 44 behavioral arousals were recorded in the laboratory; seven episodes at baseline (five from SWS, two from stage 2 sleep) and 37 episodes during recovery sleep (30 from SWS, seven from stage 2 sleep). There was no significant difference in the distribution of postarousal EEG patterns identified during baseline and recovery sleep. One pattern, comprised of diffuse rhythmic and synchronous delta activity, was preferentially associated with relatively simple behavioral episodes but did not occur during episodes from stage 2 sleep. Overall, delta activity was detected in 48% of the behavioral episodes from SWS and in 22% of those from stage 2. There was no evidence of complete awakening during any of the episodes. The results support the view of somnambulism as a disorder of arousal and suggest that sleepwalkers' atypical arousal reactions can manifest themselves in stage 2 sleep in addition to SWS. PMID- 15339265 TI - Proteases in host cell invasion by the malaria parasite. AB - The life cycle of the malaria parasite contains three distinct invasive forms, or zoites. For at least two of these--the sporozoite and the blood-stage merozoite- invasion into their respective host cell requires the activity of parasite proteases. This review summarizes the evidence for this, discusses selected well described proteolytic modifications linked to invasion, and describes recent progress towards identifying the proteases involved. PMID- 15339266 TI - Proteases in pathogenesis and plant defence. AB - Plant pathogens deliver a variety of virulence factors to host cells to suppress basal defence responses and create suitable environments for their propagation. Plants have in turn evolved disease resistance genes whose products detect the virulence factors as a signal of invasion and activate effective defence responses. Understanding how a virulence effector contributes to virulence on susceptible hosts but becomes an avirulence factor that triggers defence responses on resistance hosts has been a major focus in plant research. Recent studies have shown that a growing list of pathogen-encoded effectors functions as proteases that are secreted into plant cells to modify host proteins. In addition, several plant proteases have been found to function in activation of the defence mechanism. These findings reveal that post-translational modification of host proteins through proteolytic processing is a widely used mechanism in regulating the plant defence response. PMID- 15339267 TI - Candida albicans proteinases and host/pathogen interactions. AB - Candida infections are common, debilitating and often recurring fungal diseases and a problem of significant clinical importance. Candida albicans, the most virulent of the Candida spp., can cause severe mucosal and life-threatening systemic infections in immunocompromised hosts. Attributes that contribute to C. albicans virulence include adhesion, hyphal formation, phenotypic switching and extracellular hydrolytic enzyme production. The extracellular hydrolytic enzymes, especially the secreted aspartyl proteinases (Saps), are one of few gene products that have been shown to directly contribute to C. albicans pathogenicity. Because C. albicans is able to colonize and infect almost every tissue in the human host, it may be crucial for the fungus to possess a number of similar but independently regulated and functionally distinct secreted proteinases to provide sufficient flexibility in order to survive and promote infection at different niche sites. The aim of this review is to explore the functional roles of the C. albicans proteinases and how they may contribute to the host/pathogen interaction in vivo. PMID- 15339268 TI - Secreted proteins from Neisseria meningitidis mediate differential human gene expression and immune activation. AB - Meningococcal secreted proteins (MSPs) have been poorly characterized. We hypothesized that MSPs play essential roles in host--bacterial interactions and in the pathogenesis of disease. In order to test this, we examined differential host gene expression in human meningeal-derived cells, in response to endotoxin depleted MSPs compared to live bacteria. Using expression arrays, upregulated expression of several pro-inflammatory and apoptosis-related genes was found to be induced by MSPs. The transcription and translation of representative genes was confirmed by using various methods. Increased interleukin 8 (IL-8) and cyclooxygenase 2 (COX-2) gene transcription was confirmed using real-time PCR. Upregulated IL-8, IL-6, ICAM-1 and COX-2 protein expression were confirmed by ELISA, flow cytometry or Western immunoblots. Furthermore, exposure of cells to MSPs or live meningococci induced a small significant resistance effect to staurosporine-induced apoptosis. Secreted meningococcal virulence factors are therefore important in inducing host inflammatory responses and resistance to apoptosis, and they are worthy of extensive investigation. PMID- 15339269 TI - Transcriptome analysis and gene expression profiles of early apoptosis-related genes in Streptococcus pyogenes-infected epithelial cells. AB - Epithelial cells are the initial sites of host invasion by group A Streptococcus pyogenes (GAS), and GAS infection of epithelial cells has been suggested to induce apoptosis. We previously reported that the induction of apoptosis is strongly associated with the protein F1-mediated invasion. We present here the gene expression profiles of the human epithelial HEp-2 cells during GAS-induced apoptosis, using serial gene analysis of expression (SAGE) analysis and macroarray analysis of apoptosis-related genes. Serial gene analysis of expression revealed the downregulation of voltage-dependent anion channels 1 and 2 genes and the upregulation of the cytochrome c oxidase and calcium binding protein genes (calpactin, calgizzarin and programmed cell death 6). Macroarray analysis and quantitative RT-PCR analysis also revealed that the genes for IL 1beta, IL-12 p35, IL12 p40, and GM-CSF are also markedly induced by GAS invasion. Furthermore, caspase-1, -9, and -14 genes are significantly upregulated during GAS invasion. These observations indicated that apoptosis associated with GAS invasion is mainly induced by mitochondrial dysfunction and calcium regulation as well as by stress, and that these transcriptional controls may regulate the cellular response to GAS invasion. PMID- 15339270 TI - A synthetic peptide as a novel anticryptococcal agent. AB - An engineered, killer decapeptide (KP) has been synthesized based on the sequence of a recombinant, single-chain anti-idiotypic antibody (KT-scFv) acting as a functional internal image of a yeast killer toxin. Killer decapeptide exerted a strong fungicidal activity against Candida albicans, which was attributed to peptide interaction with beta-glucan. As this polysaccharide is also a critical component of the cryptococcal cell wall, we wondered whether KP was also active against Cryptococcus neoformans, a human pathogen of increasing medical importance. We found that KP was able to kill both capsular and acapsular C. neoformans cells in vitro. Furthermore, KP impaired the production of specific C. neoformans virulence factors including protease and urease activity and capsule formation, rendering the fungus more susceptible to natural effector cells. In vivo treatment with KP significantly reduced fungal burden in mice with cryptococcosis and, importantly, protected the majority of immunosuppressed animals from an otherwise lethal infection. Given the relevance of cryptococcosis in immunocompromised individuals and the inability of conventional drugs to completely resolve the infection, the results of the present study indicate KP as an ideal candidate for further studies on novel anticryptococcal agents. PMID- 15339271 TI - Organ specificity, colonization and clearance dynamics in vivo following oral challenges with the murine pathogen Citrobacter rodentium. AB - Citrobacter rodentium belongs to a family of human and animal enteric pathogens that includes the clinically significant enterohaemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC). These pathogens use attaching and effacing (A/E) lesions to colonize the host gastrointestinal tract. In this study we have used bioluminescence imaging (BLI) to investigate the organ specificity, dynamics of colonization and clearance of mice by C. rodentium in situ and in real time. The bioluminescent C. rodentium derivative, strain ICC180, expresses the luxCDABE operon from the entemopathogenic nematode symbiont Photorhabdus luminescens and light levels accurately reflect bacterial numbers both in vitro and in vivo. We have demonstrated that primary colonization of the mouse by C. rodentium takes place within the caecum, specifically within the specialized patch of lymphoid tissue known as the caecal patch. Following colonization of the caecum C. rodentium established a colonic infection. Clearance of C. rodentium ICC180 parallels the colonization dynamics, i.e. the caecum was first to be cleared followed by the colon. A bioluminescent eae (encoding the outer membrane adhesin intimin) C. rodentium mutant failed to establish long-term colonization, although low levels of bacteria could be recovered for up to 3 days post challenge from the caecum. PMID- 15339272 TI - Modulation of cellular phosphatidylinositol 3-phosphate levels in primary macrophages affects heat-killed but not viable Mycobacterium avium's transport through the phagosome maturation process. AB - Most disease causing mycobacteria are intramacrophage pathogens which replicate within nonacidified phagosomes that can interact with the early endosomal network but fail to mature to a phagolysosome. The mycobacterial phagosome retain some proteins required for fusion with endocytic vesicles including Rab5 but lack others such as early endosomal autoantigen 1 (EEA1). As the membrane lipid phosphatidylinositol 3-phosphate (PtdIns-3-P) is required for EEA1 membrane association and phagosome maturation, it may be a potential target of pathogenic mycobacteria. To test this hypothesis, macrophage cellular levels of PtdIns-3-P were altered by retroviral introduction of the type III Phosphoinositide 3-Kinase (VPS34) and the PtdIns-3-P phosphatase myotubularin 1 (MTM1). By utilizing the PtdIns-3-P-specific probes FYVE and PX coupled to EGFP (EGFP-2-FYVE and EGFP-PX, respectively), the expression of PtdIns-3-P on the mycobacterial phagosome was addressed. All phagosomes containing viable Mycobacterium avium stained positive for EGFP-2-FYVE and EGFP-PX despite obvious differences in PtdIns-3-P concentrations in cells expressing MTM1 or VPS34. Altering PtdIns-3-P cellular concentrations did not affect trafficking of live bacilli. However, a significant increase in the transport of killed bacilli to a late endosomal/lysosomal compartment was observed in VPS34-compared to MTM1-transduced macrophages. Therefore, although overexpression of PdtIns-3-P in macrophages can facilitate phagosome maturation, its effect on phagosomes containing viable M. avium was negligible. PMID- 15339273 TI - Cytokeratin 18 interacts with the enteropathogenic Escherichia coli secreted protein F (EspF) and is redistributed after infection. AB - Enteropathogenic Escherichia coli (EPEC) pathogenesis requires the delivery of effector proteins into host cytosol by a type III secretion system. The effector protein EspF, while critical for disruption of epithelial barrier function through alteration of tight junctions, is not required for bacterial viability or attachment. Yeast two-hybrid analyses revealed host intermediate filament (IF) protein cytokeratin 18 (CK18) as an interacting partner of EspF. This was confirmed by co-immunoprecipitation of extracts from EPEC-infected epithelial cells. EPEC infection increased detergent-soluble CK18 amounts without significantly altering CK18 expression. The adaptor protein 14-3-3 binds to CK18 and modulates its solubility. EPEC infection promoted CK18/14-3-3 interactions, corresponding to the increase of CK18 in the soluble fractions. 14-3-3 also co immunoprecipitated with EspF, suggesting that CK18, 14-3-3 and EspF may form a complex in infected cells. The 14-3-3zeta isoform was co-immunoprecipitated with CK18, suggesting the involvement of specific signalling pathways. Immunofluorescence studies revealed a dramatic alteration in the architecture of the IF network in EPEC-infected epithelial cells. IF fragmentation, evident at 2 h post infection, progressed to a collapse of this network at later time points. The secretion mutant (DeltaescN) failed to alter CK18 solubility and IF morphology, while deletion of espF partially impaired the ability of EPEC to induce CK18 modifications. These results suggest that modifications in 14-3-3 interactions and IF network, modulated by type III secreted proteins, may be an important step in EPEC pathogenesis. PMID- 15339274 TI - A Bayesian approach to DNA sequence segmentation. AB - Many deoxyribonucleic acid (DNA) sequences display compositional heterogeneity in the form of segments of similar structure. This article describes a Bayesian method that identifies such segments by using a Markov chain governed by a hidden Markov model. Markov chain Monte Carlo (MCMC) techniques are employed to compute all posterior quantities of interest and, in particular, allow inferences to be made regarding the number of segment types and the order of Markov dependence in the DNA sequence. The method is applied to the segmentation of the bacteriophage lambda genome, a common benchmark sequence used for the comparison of statistical segmentation algorithms. PMID- 15339280 TI - Two-stage designs for gene-disease association studies with sample size constraints. AB - Gene-disease association studies based on case-control designs may often be used to identify candidate polymorphisms (markers) conferring disease risk. If a large number of markers are studied, genotyping all markers on all samples is inefficient in resource utilization. Here, we propose an alternative two-stage method to identify disease-susceptibility markers. In the first stage all markers are evaluated on a fraction of the available subjects. The most promising markers are then evaluated on the remaining individuals in Stage 2. This approach can be cost effective since markers unlikely to be associated with the disease can be eliminated in the first stage. Using simulations we show that, when the markers are independent and when they are correlated, the two-stage approach provides a substantial reduction in the total number of marker evaluations for a minimal loss of power. The power of the two-stage approach is evaluated when a single marker is associated with the disease, and in the presence of multiple disease susceptibility markers. As a general guideline, the simulations over a wide range of parametric configurations indicate that evaluating all the markers on 50% of the individuals in Stage 1 and evaluating the most promising 10% of the markers on the remaining individuals in Stage 2 provides near-optimal power while resulting in a 45% decrease in the total number of marker evaluations. PMID- 15339281 TI - An extended general location model for causal inferences from data subject to noncompliance and missing values. AB - Noncompliance is a common problem in experiments involving randomized assignment of treatments, and standard analyses based on intention-to-treat or treatment received have limitations. An attractive alternative is to estimate the Complier Average Causal Effect (CACE), which is the average treatment effect for the subpopulation of subjects who would comply under either treatment (Angrist, Imbens, and Rubin, 1996, Journal of American Statistical Association 91, 444 472). We propose an extended general location model to estimate the CACE from data with noncompliance and missing data in the outcome and in baseline covariates. Models for both continuous and categorical outcomes and ignorable and latent ignorable (Frangakis and Rubin, 1999, Biometrika 86, 365-379) missing-data mechanisms are developed. Inferences for the models are based on the EM algorithm and Bayesian MCMC methods. We present results from simulations that investigate sensitivity to model assumptions and the influence of missing-data mechanism. We also apply the method to the data from a job search intervention for unemployed workers. PMID- 15339282 TI - Penalized partial likelihood for frailties and smoothing splines in time to first insemination models for dairy cows. AB - In many epidemiological studies time to event data are clustered and the physiological relationship between (time-dependent) covariates and the log hazard is often not linear as assumed in the Cox model. Introducing frailties in the Cox model can account for the clustering of the data and smoothing splines can be used to describe nonlinear relations. These two extensions of the Cox model are introduced jointly and it is shown how penalized partial likelihood techniques can be used to fit the extended model. We demonstrate the need for such a model to study the relation between the physiological covariates milk ureum and protein concentration and the log hazard of first insemination in dairy cows, with the farms as clusters. PMID- 15339283 TI - Infections with varying contact rates: application to varicella. AB - We develop methods for the analysis of infectious disease data when age-specific contact rates vary over time. Our methods are valid when contact rates vary slowly on the time scale of the infection process, and are applicable to a variety of data types including serial seroprevalence surveys and case reports. The methods exploit approximate endemic equilibria, and require numerical solution of an associated integral equation in age and time. We also estimate summary statistics such as time-dependent analogs of the basic reproduction number and critical immunization threshold. We illustrate the methods with data on varicella (chickenpox) in the United Kingdom. PMID- 15339284 TI - Maximum likelihood analysis of a general latent variable model with hierarchically mixed data. AB - A general two-level latent variable model is developed to provide a comprehensive framework for model comparison of various submodels. Nonlinear relationships among the latent variables in the structural equations at both levels, as well as the effects of fixed covariates in the measurement and structural equations at both levels, can be analyzed within the framework. Moreover, the methodology can be applied to hierarchically mixed continuous, dichotomous, and polytomous data. A Monte Carlo EM algorithm is implemented to produce the maximum likelihood estimate. The E-step is completed by approximating the conditional expectations through observations that are simulated by Markov chain Monte Carlo methods, while the M-step is completed by conditional maximization. A procedure is proposed for computing the complicated observed-data log likelihood and the BIC for model comparison. The methods are illustrated by using a real data set. PMID- 15339285 TI - Regression analysis of doubly censored failure time data using the additive hazards model. AB - Doubly censored failure time data arise when the survival time of interest is the elapsed time between two related events and observations on occurrences of both events could be censored. Regression analysis of doubly censored data has recently attracted considerable attention and for this a few methods have been proposed (Kim et al., 1993, Biometrics 49, 13-22; Sun et al., 1999, Biometrics 55, 909-914; Pan, 2001, Biometrics 57, 1245-1250). However, all of the methods are based on the proportional hazards model and it is well known that the proportional hazards model may not fit failure time data well sometimes. This article investigates regression analysis of such data using the additive hazards model and an estimating equation approach is proposed for inference about regression parameters of interest. The proposed method can be easily implemented and the properties of the proposed estimates of regression parameters are established. The method is applied to a set of doubly censored data from an AIDS cohort study. PMID- 15339286 TI - A semiparametric method for analyzing matched case-control family studies with a continuous outcome and proband sampling. AB - We consider matched case-control familial studies which match a group of patients, called "case probands," with a group of disease-free subjects, called "control probands," using a set of family-level matching variables. Family members of each proband are then recruited into the study. Of interest here is the familial aggregation of the response variable and the effects of subject specific covariates on the response. We propose an estimating equation approach to jointly estimate the main effects and intrafamilial correlations for matched family studies with a continuous outcome. Only knowledge of the first two joint moments of the response variable is required. The induced estimators for the main effects and intrafamilial correlations are consistent and asymptotically normally distributed. We apply the proposed method to sleep apnea data. A simulation study demonstrates the usefulness of our approach. PMID- 15339287 TI - Estimating the cumulative risk of a false-positive test in a repeated screening program. AB - The goal of screening tests for a chronic disease such as cancer is early detection and treatment with a consequent reduction in mortality from the disease. Screening tests, however, might produce false positive and false negative results. With an increasing number of screening tests, it is clear that the risk of a false-positive screen, a finding with potentially significant emotional, financial, and health costs, also increases. Elmore et al. (1998, New England Journal of Medicine 338, 1089-1096), Christiansen et al. (2000, Journal of the National Cancer Institute 92, 1657-1666), and Gelfand and Wang (2000, Statistics in Medicine 19, 1865-1879) investigated this problem under the somewhat unrealistic assumption that the choice of making the decision to drop out at the kth screen does not depend upon the results of the earlier k - 1 screens. In this article we obtain sufficient and necessary conditions for their assumption to hold and use one of them to provide a method for testing the validity of the assumption. A new model which does not depend on their assumption is introduced. The maximum likelihood estimator of the cumulative risk of receiving a false-positive screen under the new model is derived and its asymptotic normality is proved. The extension of the new model by incorporating covariate information is also considered. We apply our testing method and the new model to data from the breast cancer screening trial of the Health Insurance Plan of Greater New York. PMID- 15339288 TI - Designs for single- or multiple-agent phase I trials. AB - Phase I trials of cytotoxic agents in oncology are usually dose-finding studies that involve a single cytotoxic agent. Many statistical methods have been proposed for these trials, all of which are based on the assumption of a monotonic dose-toxicity curve. For single-agent trials, this is a valid assumption. In many trials, however, investigators are interested in finding the maximally tolerated dose based on escalating multiple cytotoxic agents. When there are multiple agents, monotonicity of the dose-toxicity curve is not clearly defined. In this article we present a design for phase I trials in which the toxicity probabilities follow a partial order, meaning that there are pairs of treatments for which the ordering of the toxicity probabilities is not known at the start of the trial. We compare the new design to existing methods for simple orders and investigate the properties of the design for two partial orders. PMID- 15339289 TI - Estimation of growth parameters from multiple-recapture data. AB - This article develops a method for analysis of growth data with multiple recaptures when the initial ages for all individuals are unknown. The existing approaches either impute the initial ages or model them as random effects. Assumptions about the initial age are not verifiable because all the initial ages are unknown. We present an alternative approach that treats all the lengths including the length at first capture as correlated repeated measures for each individual. Optimal estimating equations are developed using the generalized estimating equations approach that only requires the first two moment assumptions. Explicit expressions for estimation of both mean growth parameters and variance components are given to minimize the computational complexity. Simulation studies indicate that the proposed method works well. Two real data sets are analyzed for illustration, one from whelks (Dicathais aegaota) and the other from southern rock lobster (Jasus edwardsii) in South Australia. PMID- 15339290 TI - Bayesian modeling of multiple lesion onset and growth from interval-censored data. AB - In studying rates of occurrence and progression of lesions (or tumors), it is typically not possible to obtain exact onset times for each lesion. Instead, data consist of the number of lesions that reach a detectable size between screening examinations, along with measures of the size/severity of individual lesions at each exam time. This interval-censored data structure makes it difficult to properly adjust for the onset time distribution in assessing covariate effects on rates of lesion progression. This article proposes a joint model for the multiple lesion onset and progression process, motivated by cross-sectional data from a study of uterine leiomyoma tumors. By using a joint model, one can potentially obtain more precise inferences on rates of onset, while also performing onset time-adjusted inferences on lesion severity. Following a Bayesian approach, we propose a data augmentation Markov chain Monte Carlo algorithm for posterior computation. PMID- 15339291 TI - Dose-finding based on efficacy-toxicity trade-offs. AB - We present an adaptive Bayesian method for dose-finding in phase I/II clinical trials based on trade-offs between the probabilities of treatment efficacy and toxicity. The method accommodates either trinary or bivariate binary outcomes, as well as efficacy probabilities that possibly are nonmonotone in dose. Doses are selected for successive patient cohorts based on a set of efficacy-toxicity trade off contours that partition the two-dimensional outcome probability domain. Priors are established by solving for hyperparameters that optimize the fit of the model to elicited mean outcome probabilities. For trinary outcomes, the new algorithm is compared to the method of Thall and Russell (1998, Biometrics 54, 251-264) by application to a trial of rapid treatment for ischemic stroke. The bivariate binary outcome case is illustrated by a trial of graft-versus-host disease treatment in allogeneic bone marrow transplantation. Computer simulations show that, under a wide rage of dose-outcome scenarios, the new method has high probabilities of making correct decisions and treats most patients at doses with desirable efficacy-toxicity trade-offs. PMID- 15339292 TI - Tests for establishing compatibility of an observed genotype distribution with Hardy-Weinberg equilibrium in the case of a biallelic locus. AB - The classical chi(2)-procedure for the assessment of genetic equilibrium is tailored for establishing lack rather than goodness of fit of an observed genotype distribution to a model satisfying the Hardy-Weinberg law, and the same is true for the exact competitors to the large-sample procedure, which have been proposed in the biostatistical literature since the late 1930s. In this contribution, the methodology of statistical equivalence testing is adopted for the construction of tests for problems in which the assumption of approximate compatibility of the genotype distribution actually sampled with Hardy-Weinberg equilibrium (HWE) plays the role of the alternative hypothesis one aims to establish. The result of such a construction highly depends on the choice of a measure of distance to be used for defining an indifference zone containing those genotype distributions whose degree of disequilibrium shall be considered irrelevant. The first such measure proposed here is the Euclidean distance of the true parameter vector from that of a genotype distribution with identical allele frequencies being in strict HWE. The second measure is based on the (scalar) parameter of the distribution first introduced into the present context by Stevens (1938, Annals of Eugenics 8, 377-383). The first approach leads to a nonconditional test (which nevertheless can be carried out in a numerically exact way), the second to an exact conditional test shown to be uniformly most powerful unbiased (UMPU) for the associated pair of hypotheses. Both tests are compared in terms of the exact power attained against the class of those specific alternatives under which HWE is strictly satisfied. PMID- 15339293 TI - Applications of a parametric model for informative censoring. AB - In this article, we explore the use of a parametric model (for analyzing survival data) which is defined to allow sensitivity analysis for the presence of informative censoring. The dependence between the failure and the censoring processes is expressed through a parameter delta and a general bias function B(t, theta). We calculate the expectation of the potential bias due to informative censoring, which is an overall measure of how misleading our results might be if censoring is actually nonignorable. Bounds are also calculated for quantities of interest, e.g., parameter of the distribution of the failure process, which do not depend on the choice of the bias function for fixed delta. An application that relates to systematic lupus erythematosus data illustrates how additional information can result in reducing the uncertainty on estimates of the location parameter. Sensitivity analysis on a relative risk parameter is also explored. PMID- 15339294 TI - Optimal conditional error functions for the control of conditional power. AB - Ethical considerations and the competitive environment of clinical trials usually require that any given trial have sufficient power to detect a treatment advance. If at an interim analysis the available data are used to decide whether the trial is promising enough to be continued, investigators and sponsors often wish to have a high conditional power, which is the probability to reject the null hypothesis given the interim data and the alternative of interest. Under this requirement a design with interim sample size recalculation, which keeps the overall and conditional power at a prespecified value and preserves the overall type I error rate, is a reasonable alternative to a classical group sequential design, in which the conditional power is often too small. In this article two stage designs with control of overall and conditional power are constructed that minimize the expected sample size, either for a simple point alternative or for a random mixture of alternatives given by a prior density for the efficacy parameter. The presented optimality result applies to trials with and without an interim hypothesis test; in addition, one can account for constraints such as a minimal sample size for the second stage. The optimal designs will be illustrated with an example, and will be compared to the frequently considered method of using the conditional type I error level of a group sequential design. PMID- 15339295 TI - Prentice's approach and the meta-analytic paradigm: a reflection on the role of statistics in the evaluation of surrogate endpoints. AB - We put a perspective on the strengths and limitations of statistical methods for the evaluation of surrogate endpoints. Whereas using several trials overcomes some of the limitations of a single-trial framework (Prentice, 1989, Statistics in Medicine 8, 431-440), arguably the evaluation of surrogate endpoints can never be done using only statistical evidence but such evidence should be seen as but one component in a decision-making process that involves, among others, a number of clinical and biological considerations. We briefly present a hierarchical framework that incorporates ideas from Prentice's work and is uniformly applicable to different types of surrogate and true clinical outcomes. PMID- 15339296 TI - Functional mapping of quantitative trait loci underlying growth trajectories using a transform-both-sides logistic model. AB - The incorporation of developmental control mechanisms of growth has proven to be a powerful tool in mapping quantitative trait loci (QTL) underlying growth trajectories. A theoretical framework for implementing a QTL mapping strategy with growth laws has been established. This framework can be generalized to an arbitrary number of time points, where growth is measured, and becomes computationally more tractable, when the assumption of variance stationarity is made. In practice, however, this assumption is likely to be violated for age specific growth traits due to a scale effect. In this article, we present a new statistical model for mapping growth QTL, which also addresses the problem of variance stationarity, by using a transform-both-sides (TBS) model advocated by Carroll and Ruppert (1984, Journal of the American Statistical Association 79, 321-328). The TBS-based model for mapping growth QTL cannot only maintain the original biological properties of a growth model, but also can increase the accuracy and precision of parameter estimation and the power to detect a QTL responsible for growth differentiation. Using the TBS-based model, we successfully map a QTL governing growth trajectories to a linkage group in an example of forest trees. The statistical and biological properties of the estimates of this growth QTL position and effect are investigated using Monte Carlo simulation studies. The implications of our model for understanding the genetic architecture of growth are discussed. PMID- 15339297 TI - Bayesian multivariate logistic regression. AB - Bayesian analyses of multivariate binary or categorical outcomes typically rely on probit or mixed effects logistic regression models that do not have a marginal logistic structure for the individual outcomes. In addition, difficulties arise when simple noninformative priors are chosen for the covariance parameters. Motivated by these problems, we propose a new type of multivariate logistic distribution that can be used to construct a likelihood for multivariate logistic regression analysis of binary and categorical data. The model for individual outcomes has a marginal logistic structure, simplifying interpretation. We follow a Bayesian approach to estimation and inference, developing an efficient data augmentation algorithm for posterior computation. The method is illustrated with application to a neurotoxicology study. PMID- 15339298 TI - Shared frailty models for recurrent events and a terminal event. AB - There has been an increasing interest in the analysis of recurrent event data (Cook and Lawless, 2002, Statistical Methods in Medical Research 11, 141-166). In many situations, a terminating event such as death can happen during the follow up period to preclude further occurrence of the recurrent events. Furthermore, the death time may be dependent on the recurrent event history. In this article we consider frailty proportional hazards models for the recurrent and terminal event processes. The dependence is modeled by conditioning on a shared frailty that is included in both hazard functions. Covariate effects can be taken into account in the model as well. Maximum likelihood estimation and inference are carried out through a Monte Carlo EM algorithm with Metropolis-Hastings sampler in the E-step. An analysis of hospitalization and death data for waitlisted dialysis patients is presented to illustrate the proposed methods. Methods to check the validity of the proposed model are also demonstrated. This model avoids the difficulties encountered in alternative approaches which attempt to specify a dependent joint distribution with marginal proportional hazards and yields an estimate of the degree of dependence. PMID- 15339299 TI - Predicting and correcting bias caused by measurement error in line transect sampling using multiplicative error models. AB - Line transect sampling is one of the most widely used methods for animal abundance assessment. Standard estimation methods assume certain detection on the transect, no animal movement, and no measurement errors. Failure of the assumptions can cause substantial bias. In this work, the effect of error measurement on line transect estimators is investigated. Based on considerations of the process generating the errors, a multiplicative error model is presented and a simple way of correcting estimates based on knowledge of the error distribution is proposed. Using beta models for the error distribution, the effect of errors and of the proposed correction is assessed by simulation. Adequate confidence intervals for the corrected estimates are obtained using a bootstrap variance estimate for the correction and the delta method. As noted by Chen (1998, Biometrics 54, 899-908), even unbiased estimators of the distances might lead to biased density estimators, depending on the actual error distribution. In contrast with the findings of Chen, who used an additive model, unbiased estimation of distances, given a multiplicative model, lead to overestimation of density. Some error distributions result in observed distance distributions that make efficient estimation impossible, by removing the shoulder present in the original detection function. This indicates the need to improve field methods to reduce measurement error. An application of the new methods to a real data set is presented. PMID- 15339300 TI - Dynamic analysis of multivariate failure time data. AB - We present an approach for analyzing internal dependencies in counting processes. This covers the case with repeated events on each of a number of individuals, and more generally, the situation where several processes are observed for each individual. We define dynamic covariates, i.e., covariates depending on the past of the processes. The statistical analysis is performed mainly by the nonparametric additive approach. This yields a method for analyzing multivariate survival data, which is an alternative to the frailty approach. We present cumulative regression plots, statistical tests, residual plots, and a hat matrix plot for studying outliers. A program in R and S-PLUS for analyzing survival data with the additive regression model is available on the web site http://www.med.uio.no/imb/stat/addreg. The program has been developed to fit the counting process framework. PMID- 15339301 TI - Multiple-testing strategy for analyzing cDNA array data on gene expression. AB - An objective of many functional genomics studies is to estimate treatment-induced changes in gene expression. cDNA arrays interrogate each tissue sample for the levels of mRNA for hundreds to tens of thousands of genes, and the use of this technology leads to a multitude of treatment contrasts. By-gene hypotheses tests evaluate the evidence supporting no effect, but selecting a significance level requires dealing with the multitude of comparisons. The p-values from these tests order the genes such that a p-value cutoff divides the genes into two sets. Ideally one set would contain the affected genes and the other would contain the unaffected genes. However, the set of genes selected as affected will have false positives, i.e., genes that are not affected by treatment. Likewise, the other set of genes, selected as unaffected, will contain false negatives, i.e., genes that are affected. A plot of the observed p-values (1 - p) versus their expectation under a uniform [0, 1] distribution allows one to estimate the number of true null hypotheses. With this estimate, the false positive rates and false negative rates associated with any p-value cutoff can be estimated. When computed for a range of cutoffs, these rates summarize the ability of the study to resolve effects. In our work, we are more interested in selecting most of the affected genes rather than protecting against a few false positives. An optimum cutoff, i.e., the best set given the data, depends upon the relative cost of falsely classifying a gene as affected versus the cost of falsely classifying a gene as unaffected. We select the cutoff by a decision-theoretic method analogous to methods developed for receiver operating characteristic curves. In addition, we estimate the false discovery rate and the false nondiscovery rate associated with any cutoff value. Two functional genomics studies that were designed to assess a treatment effect are used to illustrate how the methods allowed the investigators to determine a cutoff to suit their research goals. PMID- 15339302 TI - Two-phase sampling for simultaneous prevalence estimation and case detection. AB - Two-phase designs for estimation of prevalence, where the first-phase classification is fallible and the second is accurate but relatively expensive, are not necessarily justified on efficiency grounds. However, they might be advantageous for dual-purpose studies, for example where prevalence estimation is followed by a clinical trial or case-control study, if they can identify cases of disease for the second study in a cost-effective way. Alternatively, they may be justified on ethical grounds if they can identify more, previously undetected but treatable cases of disease, than a simple random sample design. An approach to sampling is proposed, which formally combines the goals of efficient prevalence estimation and case detection by setting different notional study costs for investigating cases and noncases. Two variants of the method are compared with an "ethical" two-phase scheme proposed by Shrout and Newman (1989, Biometrics 45, 549-555), and with the most efficient scheme for prevalence estimation alone, in terms of the standard error of the prevalence estimate, the expected number of cases, and the fraction of cases among second-phase subjects, given a fixed budget. One variant yields the highest fraction and expected number of cases but also the largest standard errors. The other yields a higher fraction than Shrout and Newman's scheme and a similar number of cases but appears to do so more efficiently. PMID- 15339303 TI - A parametric model for studying organism fitness using step-stress experiments. AB - We propose a method based on parametric survival analysis to analyze step-stress data. Step-stress studies are failure time studies in which the experimental stressor is increased at specified time intervals. While this protocol has been frequently employed in industrial reliability studies, it is less common in the life sciences. Possible biological applications include experiments on swimming performance of fish using a step function defining increasing water velocity over time, and treadmill tests on humans. A likelihood-ratio test is developed for comparing the failure times in two groups based on a piecewise constant hazard assumption. The test can be extended to other piecewise distributions and to include covariates. An example data set is used to illustrate the method and highlight experimental design issues. A small simulation study compares this analysis procedure to currently used methods with regard to type I error rate and power. PMID- 15339304 TI - Calculation of sample size in survival trials: the impact of informative noncompliance. AB - Sample size calculations for survival trials typically include an adjustment to account for the expected rate of noncompliance, or discontinuation from study medication. Existing sample size methods assume that when patients discontinue, they do so independently of their risk of an endpoint; that is, that noncompliance is noninformative. However, this assumption is not always true, as we illustrate using results from a published clinical trial database. In this article, we introduce a modified version of the method proposed by Lakatos (1988, Biometrics 44, 229-241) that can be used to calculate sample size under informative noncompliance. This method is based on the concept of two subpopulations: one with high rates of endpoint and discontinuation and another with low rates. Using this new method, we show that failure to consider the impact of informative noncompliance can lead to a considerably underpowered study. PMID- 15339305 TI - Confidence interval estimation of the intraclass correlation coefficient for binary outcome data. AB - We obtain closed-form asymptotic variance formulae for three point estimators of the intraclass correlation coefficient that may be applied to binary outcome data arising in clusters of variable size. Our results include as special cases those that have previously appeared in the literature (Fleiss and Cuzick, 1979, Applied Psychological Measurement 3, 537-542; Bloch and Kraemer, 1989, Biometrics 45, 269 287; Altaye, Donner, and Klar, 2001, Biometrics 57, 584-588). Simulation results indicate that confidence intervals based on the estimator proposed by Fleiss and Cuzick provide coverage levels close to nominal over a wide range of parameter combinations. Two examples are presented. PMID- 15339306 TI - Bayesian variable selection in multinomial probit models to identify molecular signatures of disease stage. AB - Here we focus on discrimination problems where the number of predictors substantially exceeds the sample size and we propose a Bayesian variable selection approach to multinomial probit models. Our method makes use of mixture priors and Markov chain Monte Carlo techniques to select sets of variables that differ among the classes. We apply our methodology to a problem in functional genomics using gene expression profiling data. The aim of the analysis is to identify molecular signatures that characterize two different stages of rheumatoid arthritis. PMID- 15339307 TI - Marginal analysis of incomplete longitudinal binary data: a cautionary note on LOCF imputation. AB - In recent years there has been considerable research devoted to the development of methods for the analysis of incomplete data in longitudinal studies. Despite these advances, the methods used in practice have changed relatively little, particularly in the reporting of pharmaceutical trials. In this setting, perhaps the most widely adopted strategy for dealing with incomplete longitudinal data is imputation by the "last observation carried forward" (LOCF) approach, in which values for missing responses are imputed using observations from the most recently completed assessment. We examine the asymptotic and empirical bias, the empirical type I error rate, and the empirical coverage probability associated with estimators and tests of treatment effect based on the LOCF imputation strategy. We consider a setting involving longitudinal binary data with longitudinal analyses based on generalized estimating equations, and an analysis based simply on the response at the end of the scheduled follow-up. We find that for both of these approaches, imputation by LOCF can lead to substantial biases in estimators of treatment effects, the type I error rates of associated tests can be greatly inflated, and the coverage probability can be far from the nominal level. Alternative analyses based on all available data lead to estimators with comparatively small bias, and inverse probability weighted analyses yield consistent estimators subject to correct specification of the missing data process. We illustrate the differences between various methods of dealing with drop-outs using data from a study of smoking behavior. PMID- 15339308 TI - Some unexamined aspects of analysis of covariance in pretest-posttest studies. AB - The use of an analysis of covariance (ANCOVA) model in a pretest-posttest setting deserves to be studied separately from its use in other (non-pretest-posttest) settings. For pretest-posttest studies, the following points are made in this article: (a) If the familiar change from baseline model accurately describes the data-generating mechanism for a randomized study then it is impossible for unequal slopes to exist. Conversely, if unequal slopes exist, then it implies that the change from baseline model as a data-generating mechanism is inappropriate. An alternative data-generating model should be identified and the validity of the ANCOVA model should be demonstrated. (b) Under the usual assumptions of equal pretest and posttest within-subject error variances, the ratio of the standard error of a treatment contrast from a change from baseline analysis to that from ANCOVA is less than 2(1)/(2). (c) For an observational study it is possible for unequal slopes to exist even if the change from baseline model describes the data-generating mechanism. (d) Adjusting for the pretest variable in observational studies may actually introduce bias where none previously existed. PMID- 15339309 TI - Comment on "Estimation of population attributable fractions from fitted incidence ratios and exposure survey data, with an application to electromagnetic fields and childhood leukemia.". PMID- 15339320 TI - Systematic review: has disease outcome in Crohn's disease changed during the last four decades? AB - BACKGROUND: Disease outcome in Crohn's disease might have changed during the last four decades. Disease outcome measurement in Crohn's disease has methodological difficulties because of patient selection and lack of proper definition of diagnostic and outcome measurement criteria. AIM: To assess possible changes in disease outcome in Crohn's disease during the last four decades. METHODS: A systematic literature search was performed using the MEDLINE search engine and major international conference libraries. Articles and abstracts were selected according to stringent inclusion criteria. RESULTS: Forty articles and nine abstracts complied with the inclusion criteria. Seven studies with a median follow-up time between 11.1 and 17 years showed standard mortality ratios in Crohn's disease ranging between 2.16 and 0.72 with a tendency of decline during the last four decades. One study with 11.4 years mean follow-up time showed a statistically significant increased relative risk for colorectal cancer that was not confirmed by three others. Sixteen publications applied in the disease recurrence category. Probability of first resective surgery ranged between 38 and 96% during the first 15 years after diagnosis. The overall recurrence and surgical recurrence rates after first resective surgery ranged between 50 and 60, and 28 and 45% respectively during the following 15 years without an apparent time trend. CONCLUSION: This structured literature review provides no hard evidence for change in disease outcome in Crohn's disease during the last four decades. PMID- 15339321 TI - Relationship between body mass index, diet, exercise and gastro-oesophageal reflux symptoms in a community. AB - BACKGROUND: Body mass index (BMI) is a risk factor for gastro-oesophageal reflux but may simply be explained by diet and lifestyle. AIM: We aimed to determine the contribution of BMI, diet and exercise to GER. METHODS: Community subjects (n = 211, mean age = 36 years, 43% males) completed validated questionnaires on gastro oesophageal reflux, energy expenditure (Harvard Alumni Activity Survey), dietary intake (Harvard Food Frequency Questionnaire) and measures of personality and life event stress. Diet, exercise, BMI and other potential risk factors for reflux were analysed using logistic regression analyses. RESULTS: The overall mean (+/- s.d.) BMI was 26.6 (+/- 5.7); 79 (37%) reported infrequent (< weekly) reflux and 16 (8%) reported frequent (> or = weekly) reflux. The median caloric intake was 2097 cal/day and the median daily energy expenditure was 1753 cal/day. Among those with BMI > 25, 10% reported frequent reflux compared to 4% of those with BMI < or = 25. In a model which included age, sex and Symptom Checklist-90 somatisation T-score, BMI was associated with reflux (OR per 5 units = 1.9, 95% CI: 1.2, 3.0). In models which included diet and exercise variables, BMI but not diet or exercise was associated with reflux. CONCLUSION: BMI may be associated with symptomatic gastro-oesophageal reflux independent of diet and exercise. PMID- 15339322 TI - Projections of demand and capacity for colonoscopy related to increasing rates of colorectal cancer screening in the United States. AB - BACKGROUND: There is debate about the optimal colorectal cancer screening test, partly because of concerns about colonoscopy demand. AIM: To quantify the demand for colonoscopy with different screening tests, and to estimate the ability of the United States health care system to meet demand. METHODS: We used a previously published Markov model and the United States census data to estimate colonoscopy demand. We then used an endoscopic database to compare current rates of screening-related colonoscopy with those projected by the model, and to estimate the number of endoscopists needed to meet colonoscopy demand. RESULTS: Annual demand for colonoscopy ranges from 2.21 to 7.96 million. Based on current practice patterns, demand exceeds current supply regardless of screening strategy. We estimate that an increase of at least 1360 gastroenterologists would be necessary to meet demand for colonoscopic screening undergone once at age 65, while colonoscopy every 10 years could require 32 700 more gastroenterologists. A system using dedicated endoscopists could meet demand with fewer endoscopists. CONCLUSIONS: Colorectal cancer screening leads to demand for colonoscopy that outstrips supply. Systems to train dedicated screening endoscopists may be necessary in order to provide population-wide screening. The costs and feasibility of establishing this infrastructure should be studied further. PMID- 15339323 TI - Effect of acid on duodenal blood flow and mucus secretion measured by reflectance spectrophotometry: a prospective, randomized-controlled study. AB - BACKGROUND: In animals, hydrochloric acid increases blood flow and mucus secretion in the duodenal mucosa. A significant correlation between index of haemoglobin oxygen saturation and mucosal blood flow, and between change in index of haemoglobin concentration and mucus thickness, respectively, has been demonstrated by reflectance spectrophotometry. AIM: To examine the effect of topical hydrochloric acid upon mucosal blood flow and mucus secretion in the human duodenum. METHODS: This prospective study of 120 patients undergoing routine upper endoscopy, examined the effect of topical 0.1 n hydrochloric acid or 0.9% saline on the duodenal bulb in a randomized, double-blind fashion. Duodenal mucosal index of haemoglobin oxygen saturation and index of haemoglobin concentration were measured by endoscopic reflectance spectrophotometry before and after hydrochloric acid or saline. RESULTS: Baseline index of haemoglobin oxygen saturation, calculated blood flow and index of haemoglobin concentration measurements were comparable between hydrochloric acid (n = 60) and saline (n = 60) treated groups. A history of current use of non-steroidal anti-inflammatory drug was associated with a significantly lower baseline index of haemoglobin oxygen saturation and calculated blood flow. Hydrochloric acid resulted in a significant increase in index of haemoglobin oxygen saturation and calculated blood flow, but a decrease in index of haemoglobin concentration, reflecting an increase in mucus thickness compared with saline. CONCLUSIONS: Our observations in humans confirm data in animal studies that topical exposure to hydrochloric acid induces an increase in duodenal mucosal blood flow and mucus secretion. Post hoc analysis of the data also revealed that attenuation of basal duodenal mucosal blood flow is associated with a history of current non-steroidal anti inflammatory drug use. Endoscopic reflectance spectrophotometry appears to be adequate to assess factors that influence duodenal defence mechanisms of blood flow and mucus secretion in humans. PMID- 15339324 TI - Reduced incidence of upper gastrointestinal ulcer complications with the COX-2 selective inhibitor, valdecoxib. AB - AIM: In a predefined analysis, data were pooled from eight blinded, randomized, controlled trials, and separately from three long-term, open-label trials to determine the rate of upper gastrointestinal ulcer complications with the cyclo oxygenase-2 selective inhibitor, valdecoxib, vs. non-selective non-steroidal anti inflammatory drugs. METHODS: In randomized, controlled trials, 7434 osteoarthritis and rheumatoid arthritis patients received placebo (n = 973), valdecoxib 5-80 mg daily (n = 4362), or a non-selective non-steroidal anti inflammatory drug (naproxen, ibuprofen or diclofenac; n = 2099) for 12-26 weeks. In long-term, open-label trials, 2871 patients received valdecoxib 10-80 mg daily for up to 1 year. All potential events were reviewed by a blinded, independent review committee based on a priori definitions of ulcer complications (perforations, obstructions, bleeds). RESULTS: In randomized, controlled trials, 19 of 955 potential events were adjudicated to be ulcer complications. Valdecoxib was associated with a significantly lower ulcer complication rate than non selective non-steroidal anti-inflammatory drugs (0.68% vs. 1.96%, all patients; 0.29% vs. 2.08%, non-aspirin users; P < 0.05). In long-term, open-label trials, seven of 310 potential events were adjudicated to be ulcer complications; the annualized incidence for valdecoxib was 0.39% (seven of 1791 patient-years) for all patients and 0.2% (three of 1472 patient-years) for non-aspirin users. CONCLUSIONS: Valdecoxib, including above recommended doses, is associated with a significantly lower rate of upper gastrointestinal ulcer complications than therapeutic doses of non-selective non-steroidal anti-inflammatory drugs. PMID- 15339325 TI - Adjuvant biofeedback following anal sphincter repair: a randomized study. AB - AIMS: To evaluate the impact of adjuvant biofeedback following sphincter surgery. METHODS: Thirty-eight patients were randomized into sphincter repair or sphincter repair plus biofeedback groups. Outcome measures included a symptom questionnaire, patient's rating of satisfaction with continence function and improvement, change in continence score, quality of life and anorectal physiology. Endoanal ultrasonography was also performed pre- and post operatively. RESULTS: Immediately following surgery, there was no statistically significant difference in any of the functional or physiological variables between the groups. Continence and patient satisfaction scores improved with a mean difference of -0.48 (95% CI: -3.30-2.33, P = 0.73) and 1.03 (95% CI: -1.40 3.46, P = 0.39), respectively. Only the difference in embarrassment scores reached statistical significance (mean) 0.56 (95% CI: 0.12-0.99, P = 0.014). Resting and squeeze pressures also improved. Thirteen of 14 in the biofeedback and 11 of 17 (control) reported symptomatic improvement. In the biofeedback group, although not statistically significant continence and satisfaction scores improved and were sustained over time. In the control group, continence and satisfaction scores changed little between 3 and 12 months (P = NS). Quality of life measures improved within the biofeedback group but there was no statistical difference between the groups. CONCLUSION: Following surgery continence function improves in all patients but adjuvant biofeedback therapy improves quality of life and maintains symptomatic improvement over time. PMID- 15339326 TI - Fracture risk of women with primary biliary cirrhosis: no increase compared with general population controls. AB - AIM: Patients with primary biliary cirrhosis may be at increased risk of osteoporosis but to what extent this is reflected in an increased fracture risk is unknown. We have enquired about the fracture experience of female primary biliary cirrhosis patients compared with sex- and age-matched controls. METHODS: Patients aged 30-75 with primary biliary cirrhosis and age-matched controls were sent a postal questionnaire asking about their fracture history and details of risk factors for osteoporosis. RESULTS: 85 eligible patients with primary biliary cirrhosis and 116 controls responded. Forty-one per cent of patients with primary biliary cirrhosis and 30% of controls reported ever having had a fracture odds ratio 1.5 (95% confidence interval: 0.80-2.89). Twenty-eight per cent of primary biliary cirrhosis patients and 23.3% of controls reported a fracture after the age of 30, odds ratio 1.2 (95% confidence interval: 0.57-2.56), and 14.1% of primary biliary cirrhosis patients and 12.1% of controls reported a low impact fracture of the long bones or of the vertebrae odds ratio 1.0 (95% confidence interval: 0.31-2.68). CONCLUSIONS: No overall increased fracture risk in patients with primary biliary cirrhosis was observed. As a group, unselected patients with primary biliary cirrhosis do not represent a population at particularly high risk of osteoporotic fracture and thus targeting them for osteoporosis screening and treatment is not justified. Further work investigating subgroups of patients with primary biliary cirrhosis at potentially high risk of osteoporosis, such as those with advanced disease or severe cholestasis is required. PMID- 15339327 TI - Helicobacter pylori infection prevents the occurrence of the tolerance phenomenon of histamine H2 receptor antagonists. AB - BACKGROUND: The attenuated anti-secretory activity of H2 receptor antagonists (H2RA) during continuous administration is referred to as the tolerance phenomenon. However, it is not clarified whether Helicobacter pylori infection affects the occurrence of tolerance to H2RA. It is also not clarified whether the tolerance phenomenon occurs to a new H2RA, lafutidine. AIM: To investigate the occurrence of the tolerance phenomenon in subjects with and without H. pylori infection during the continuous administration of lafutidine and famotidine. SUBJECTS AND METHODS: Subjects were 20 healthy male volunteers (seven H. pylori positive and 13 H. pylori negative cases). All subjects were examined by ambulatory intragastric pH monitoring five times without medication, on the first and 15th day of the administration of 20 mg b.d. famotidine and 10 mg b.d. lafutidine in a cross-over fashion. RESULTS: The tolerance phenomenon was not observed in H. pylori-positive subjects during the 15-day-long administration of both H2RAs. In contrast, the tolerance phenomenon was observed in H. pylori negative subjects, which has been previously reported. CONCLUSIONS: This study demonstrated that H. pylori infection affects the tolerance phenomenon during continuous administration of H2RAs. PMID- 15339328 TI - Prevention of erosive oesophagitis relapse with pantoprazole. AB - AIM: To compare the safety and efficacy of pantoprazole and ranitidine in maintaining erosive oesophagitis healing. METHODS: Gastro-oesophageal reflux disease patients (349) with endoscopically documented healed erosive oesophagitis (grade 0 or 1) were randomly assigned to receive pantoprazole (10, 20 or 40 mg/q.d.s.) or ranitidine (150 mg/b.d.). Erosive oesophagitis status was assessed endoscopically at months 1, 3, 6 and 12 or when relapse symptoms appeared (relapse = reappearance of erosive oesophagitis grade 2 within 12 months). Symptom-free days were also assessed. RESULTS: Pantoprazole 20- and 40-mg were significantly more effective than ranitidine in maintaining healing regardless of initial erosive oesophagitis grade. Response was dose-related. After 12 months 78, 55, 46 and 21% of patients remained healed (40-, 20-, 10-mg pantoprazole and ranitidine). Pantoprazole 40-mg produced significantly more symptom-free days (83%) than ranitidine (58%). Heartburn-free days/nights were significantly higher with pantoprazole 40-mg (92 and 93%) than ranitidine (73 and 77%). The most frequent reason for discontinuation, unsatisfactory efficacy, occurred most often with ranitidine (P < 0.001). CONCLUSION: Once-daily pantoprazole therapy prevented relapse of healed erosive oesophagitis more effectively than ranitidine and with fewer heartburn days. Response to pantoprazole was dose-related. Pantoprazole 40-mg was the most effective regimen and consistent in maintaining erosive oesophagitis healing with a good safety and tolerability profile. PMID- 15339329 TI - Use of anti-secretory medication: a population-based cohort study. AB - BACKGROUND: Total use of anti-secretory medication (H2 blockers and proton pump inhibitors) is increasing rapidly, but knowledge of factors related to the increasing use is sparse. AIM: To describe development in the use of anti secretory medication between 1993 and 2002 at a population level. METHODS: We extracted data on use of anti-secretory medication (H2-blockers and proton pump inhibitors) and ulcerogenic drugs, demographic data, and data on gastroscopy and endoscopically verified oesophagitis and peptic ulcer diagnoses, from three large population-based databases covering the County of Funen, Denmark 1990-2002 (population 470,000). RESULTS: Between 1993 and 2002 incidence of first time users was stable at 16.7/1000 persons/year. Total amount of consumed anti secretory medication increased from 10.5 DDD/1000 persons/day to 25.2 DDD/1000 persons/day. Ninety per cent of the increase was related to long-term use of anti secretory medication (> or = 180 DDD/patient/year). In 1993 21% of the anti secretory medication was used by patients with oesophagitis, this increased to 28% in 2002. The proportion of medication used by peptic ulcer patients decreased from 29% in 1993 to 19% in 2002. CONCLUSIONS: Total use of anti-secretory medication increased as a result of more extensive long-term use, and most of the medication was used by patients without diagnosed peptic ulcer or oesophagitis. PMID- 15339330 TI - Basic fibroblast growth factor as a response parameter to infliximab in fistulizing Crohn's disease. AB - BACKGROUND: Fibroblast growth factors play an important role in (patho)physiological processes such as wound healing and tissue repair. We previously showed that basic fibroblast growth factor is actively involved in inflammatory bowel disease processes. In the present retrospective study, we assessed whether serum basic fibroblast growth factor levels in Crohn's disease patients reflect the response to anti-tumour necrosis factor-alpha antibody infliximab treatment. AIM AND METHODS: Serum samples, biopsies and patient data from a subgroup of patients included in two placebo-controlled trials were used. Fistulizing Crohn's disease patients (n = 42) were administered placebo or infliximab intravenously three times and evaluated for response up to 18 weeks. Biopsies from a subgroup of patients were stained for basic fibroblast growth factor using indirect immunohistochemistry. In the active Crohn's disease trial, patients (n = 24) received either placebo or infliximab once, and disease activity and serum basic fibroblast growth factor were assessed at weeks 0 and 4. RESULTS: Basic fibroblast growth factor levels at inclusion were comparable in the fistulizing Crohn's disease patients regardless of whether the fistulas did or did not respond or completely heal (median range: 9.3-10.6 pg/mL). At the end of follow-up basic fibroblast growth factor levels were lower in patients who responded (9.2 pg/mL, P = 0.06) or who were completely healed (8.9 pg/mL, P = 0.009) when compared with patients did not respond/heal (14.5 pg/mL), the latter not significantly increased from baseline. Decreases in the perianal disease activity index and open fistula scores at the end of the follow-up were significantly correlated with the decrease in basic fibroblast growth factor (R = 0.41; P = 0.012 and R = 0.35; P =0.027, respectively). Immunohistological evaluation also showed a trend towards decreased basic fibroblast growth factor expression in intestinal biopsies of these patients. Patients with active disease, i.e. a Crohn's disease activity index > or = 220 combined from the two studies, were found to have significantly (P = 0.0046) lower baseline serum basic fibroblast growth factors levels than those with inactive disease (5.3 vs. 10.3 pg/mL, respectively). Treatment of the active disease patients did not affect the serum basic fibroblast growth factor level, although a general decrease in disease activity was observed with infliximab treatment. CONCLUSIONS: Healing of fistulizing/perianal Crohn's disease seems to be reflected by a decrease in high serum basic fibroblast growth factor. Basic fibroblast growth factor levels do not relate with response in active Crohn's disease patients. PMID- 15339331 TI - A rational treatment of Mendelian genetics. AB - BACKGROUND: The key to a rational treatment of elementary Mendelian genetics, specifically to an understanding of the origin of dominant and recessive traits, lies in the facts that: (1) alleles of genes encode polypeptides; (2) most polypeptides are catalysts, i.e. enzymes or translocators; (3) the molecular components of all traits in all cells are the products of systems of enzymes, i.e. of fluxing metabolic pathways; (4) any flux to the molecular components of a trait responds non-linearly (non-additively) to graded mutations in the activity of any one of the enzymes at a catalytic locus in a metabolic system; (5) as the flux responds to graded changes in the activity of an enzyme, the concentrations of the molecular components of a trait also change. CONCLUSIONS: It is then possible to account rationally, and without misrepresenting Mendel, for: the origin of dominant and recessive traits; the occurrence of Mendel's 3(dominant):1(recessive) trait ratio; deviations from this ratio; the absence of dominant and recessive traits in some circumstances, the occurrence of a blending of traits in others; the frequent occurrence of pleiotropy and epistasis. PMID- 15339332 TI - Recurrence of primary extramedullary plasmacytoma in breast both simulating primary breast carcinoma. AB - BACKGROUND: Extramedullary myelomas (plasmacytoma) are malignant proliferations of plasma cells in the absence of bone involvement. When they occur in the soft tissue they usually involve the upper respiratory tract and oral cavity. Extramedullary plasmacytomas of breast are uncommon. CASE PRESENTATION: A 70 year old woman with bilateral breast masses underwent excisional biopsy for suspected primary carcinoma that subsequently proved to be a recurrence from extramedullary plasmacytoma of the mediastinum. This was diagnosed and treated 5-years prior to appearance of breast lumps. CONCLUSION: Though uncommon, considering the possibility of metastatic carcinoma and primary, secondary or recurrent lymphoproliferative disease presenting as a breast mass may avoid unnecessary surgeries. PMID- 15339333 TI - Sodium nitroprusside and peroxynitrite effect on hepatic DNases: an in vitro and in vivo study. AB - BACKGROUND: It has been documented that nitric oxide (NO) donor sodium nitroprusside (SNP) and authentic peroxynitrite are capable of promoting apoptosis in a number of different cell types. Various endonucleases have been proposed as candidates responsible for the internucleosomal cleavage of the genomic DNA observed during apoptosis, but the main effect is attributed to the alkaline-DNases (Mg2+- and caspase-dependent) and acid-DNase. The aim of this study was to examine an in vivo and in vitro possibility for alkaline- and acid DNases to be activated by SNP and peroxynitrite. RESULTS: The effect on liver tissue alkaline and acid DNase activity together with the markers of tissue and plasma oxidative and nitrosative stress (lipid peroxidation, SH group content, carbonyl groups and nitrotyrosine formation) was investigated in plasma and liver tissue. The activity of liver alkaline DNase increased and that of acid DNase decreased after in vivo treatment with either SNP or peroxynitrite. A difference observed between the in vivo and in vitro effect of oxide donor (i.e., SNP) or peroxynitrite upon alkaline DNase activity existed, and it may be due to the existence of the "inducible" endonuclease. After a spectrophotometric scan analysis of purified DNA, it was documented that both SNP and peroxynitrite induce various DNA modifications (nitroguanine formation being the most important one) whereas DNA fragmentation was not significantly increased. CONCLUSION: Alkaline DNase activation seems to be associated with the programmed destruction of the genome, leading to the fragmentation of damaged DNA sites. Thus, the elimination of damaged cells appears to be a likely factor in prevention against mutation and carcinogenesis. PMID- 15339334 TI - Leukaemia incidence among workers in the shoe and boot manufacturing industry: a case-control study. AB - BACKGROUND: Previous reports have indicated an excess of leukaemia in Broome County, New York, particularly in the Town of Union. Surveillance of cancer incidence data indicates that a large proportion of these cases occurred among males ages 65 and older. Shoe and boot manufacturing has been the largest single industry in this area throughout much of the past century. Occupational studies from Europe suggest a link between leukaemia and employment in the shoe and boot manufacturing industry. However, researchers have not found a positive association between leukaemia and employment in the shoe industry among workers in the United States. METHODS: A matched case-control study was conducted to investigate the association between leukaemia incidence among males 65 and older and employment in the shoe and boot manufacturing industry. Thirty-six cases of leukaemia occurring between 1981-1990; among males age 65 and older; residing in the town of Union met the study case criteria. Death certificates were obtained for each of the cases. These were matched to death certificates of 144 controls on date of death and date of birth +/- 1 year. Death certificates were then examined to determine the employer and occupation of each study subject. Conditional logistic regression was used to determine the risk of leukaemia among those working in the industry. RESULTS: The risk of both leukaemia (OR = 1.47; 95% CI 0.70, 3.09) and acute myeloid leukaemia (OR = 1.19; 95% CI 0.33, 4.28) were elevated among those employed in the shoe and boot manufacturing industry, however neither was statistically significant. CONCLUSION: The results, though suggestive of an association between leukaemia and employment in the shoe and boot manufacturing industry, were not statistically conclusive due mainly to limited study power. Several additional limitations may also have prevented the observance of more conclusive findings. Better exposure assessment, information on length of exposure and types of job held, control of confounding factors and information on chemicals used by this company would strengthen any future investigation. PMID- 15339335 TI - Facilitating arrhythmia simulation: the method of quantitative cellular automata modeling and parallel running. AB - BACKGROUND: Many arrhythmias are triggered by abnormal electrical activity at the ionic channel and cell level, and then evolve spatio-temporally within the heart. To understand arrhythmias better and to diagnose them more precisely by their ECG waveforms, a whole-heart model is required to explore the association between the massively parallel activities at the channel/cell level and the integrative electrophysiological phenomena at organ level. METHODS: We have developed a method to build large-scale electrophysiological models by using extended cellular automata, and to run such models on a cluster of shared memory machines. We describe here the method, including the extension of a language-based cellular automaton to implement quantitative computing, the building of a whole-heart model with Visible Human Project data, the parallelization of the model on a cluster of shared memory computers with OpenMP and MPI hybrid programming, and a simulation algorithm that links cellular activity with the ECG. RESULTS: We demonstrate that electrical activities at channel, cell, and organ levels can be traced and captured conveniently in our extended cellular automaton system. Examples of some ECG waveforms simulated with a 2-D slice are given to support the ECG simulation algorithm. A performance evaluation of the 3-D model on a four node cluster is also given. CONCLUSIONS: Quantitative multicellular modeling with extended cellular automata is a highly efficient and widely applicable method to weave experimental data at different levels into computational models. This process can be used to investigate complex and collective biological activities that can be described neither by their governing differentiation equations nor by discrete parallel computation. Transparent cluster computing is a convenient and effective method to make time-consuming simulation feasible. Arrhythmias, as a typical case, can be effectively simulated with the methods described. PMID- 15339336 TI - Wireless local area network in a prehospital environment. AB - BACKGROUND: Wireless local area networks (WLANs) are considered the next generation of clinical data network. They open the possibility for capturing clinical data in a prehospital setting (e.g., a patient's home) using various devices, such as personal digital assistants, laptops, digital electrocardiogram (EKG) machines, and even cellular phones, and transmitting the captured data to a physician or hospital. The transmission rate is crucial to the applicability of the technology in the prehospital setting. METHODS: We created two separate WLANs to simulate a virtual local are network environment such as in a patient's home or an emergency room (ER). The effects of different methods of data transmission, number of clients, and roaming among different access points on the file transfer rate were determined. RESULTS: The present results suggest that it is feasible to transfer small files such as patient demographics and EKG data from the patient's home to the ER at a reasonable speed. Encryption, user control, and access control were implemented and results discussed. CONCLUSIONS: Implementing a WLAN in a centrally managed and multiple-layer-controlled access control server is the key to ensuring its security and accessibility. Future studies should focus on product capacity, speed, compatibility, interoperability, and security management. PMID- 15339337 TI - Long acting beta2 agonists for stable chronic obstructive pulmonary disease with poor reversibility: a systematic review of randomised controlled trials. AB - BACKGROUND: The long acting beta2-agonists, salmeterol and formoterol, have been recommended, by some, as first line treatment of stable chronic obstructive pulmonary disease (COPD). We reviewed evidence of efficacy and safety when compared with placebo or anticholinergic agents in patients with poorly reversible COPD. METHODS: After searching MEDLINE, EMBASE, HealthSTAR, BIOSIS Previews, PASCAL, ToxFile, SciSearch, the Cochrane Library, and PubMed, as well as Web sites, selected journals, reference lists, and contacting drug manufacturers, two reviewers independently screened reports of randomised controlled trials of parallel or crossover design lasting four weeks or longer and including patients with a forced expiratory volume in one second (FEV1) < or = 75% of predicted, a ratio of FEV1 to forced vital capacity (FVC) < or = 88% of predicted, and < 15% improvement from baseline FEV1 after a dose of a beta2 agonist. We included trials comparing salmeterol or formoterol with placebo or with ipratropium bromide and reporting one of these outcomes: lung function; exercise capacity; quality of life scores; dyspnea; exacerbations; rescue inhaler use; incidence of tachycardia, hypokalemia, or dry mouth. Two reviewers assessed the quality of included reports using the Jadad scale and allocation concealment, and abstracted data. RESULTS: Twelve trials satisfied our inclusion criteria; eight were high quality (Jadad score >2) and four were low quality (< or = 2). The adequacy of allocation concealment was unclear in all of them. We did not perform a meta-analysis due to differences in trial design and how outcomes were reported.Two trials comparing salmeterol with ipratropium did not detect differences; one trial comparing formoterol and ipratropium described greater improvement with formoterol in morning PEFR (15.3 versus 7.1 l/min, p = 0.040). Of twelve trials comparing long acting beta2 agonists with placebo, six reported no improvement in exercise capacity, eleven reported improvements in FEV1 lung function (one reported no improvement), six reported less rescue inhaler usage (one reported no difference) and five reported improved dyspnea scores (two reported no improvement). Differences in quality of life were detected in one salmeterol trial; however, two salmeterol, and one formoterol trial reported no differences. Adverse effects of interest were not reported. CONCLUSION: In terms of clinical outcomes and safety, we could not find convincing evidence that salmeterol and formoterol have demonstrated advantages to ipratropium, a less expensive drug, for patients with stable COPD and poor reversibility. Compared to placebo, we found evidence of reduced rescue inhaler usage and improved spirometric outcomes without a significant impact on quality of life or exercise capacity. PMID- 15339338 TI - Syndromes with congenital brittle bones. AB - BACKGROUND: There is no clear definition of osteogenesis imperfecta (OI). The most widely used classification of OI divides the disease in four types, although it has been suggested that there may be at least 12 forms of OI. These forms have been named with numbers, eponyms or descriptive names. Some of these syndromes can actually be considered congenital forms of brittle bones resembling OI (SROI). DISCUSSION: A review of different syndromes with congenital brittle bones published in the literature is presented. Syndromes are classified in "OI" (those secondary to mutations in the type I pro-collagen genes), and "syndromes resembling OI" (those secondary to mutations other that the type I pro-collagen genes, identified or not). A definition for OI is proposed as a syndrome of congenital brittle bones secondary to mutations in the genes codifying for pro collagen genes (COL1A1 and COL1A2). SUMMARY: A debate about the definition of OI and a possible clinical and prognostic classification are warranted. PMID- 15339339 TI - Protein p16 as a marker of dysplastic and neoplastic alterations in cervical epithelial cells. AB - BACKGROUND: Cervical carcinomas are second most frequent type of women cancer. Success in diagnostics of this disease is due to the use of Pap-test (cytological smear analysis). However Pap-test gives significant portion of both false positive and false-negative conclusions. Amendments of the diagnostic procedure are desirable. Aetiological role of papillomaviruses in cervical cancer is established while the role of cellular gene alterations in the course of tumor progression is less clear. Several research groups including us have recently named the protein p16INK4a as a possible diagnostic marker of cervical cancer. To evaluate whether the specificity of p16INK4a expression in dysplastic and neoplastic cervical epithelium is sufficient for such application we undertook a broader immunochistochemical registration of this protein with a highly p16INK4a specific monoclonal antibody. METHODS: Paraffin-embedded samples of diagnostic biopsies and surgical materials were used. Control group included vaginal smears of healthy women and biopsy samples from patients with cervical ectopia. We examined 197 samples in total. Monoclonal antibody E6H4 (MTM Laboratories, Germany) was used. RESULTS: In control samples we did not find any p16INK4a positive cells. Overexpression of p16INK4a was detected in samples of cervical dysplasia (CINs) and carcinomas. The portion of p16INK4a-positive samples increased in the row: CIN I - CIN II - CIN III - invasive carcinoma. For all stages the samples were found to be heterogeneous with respect to p16INK4a expression. Every third of CINs III and one invasive squamous cell carcinoma (out of 21 analyzed) were negative. CONCLUSIONS: Overexpression of the protein p16INK4a is typical for dysplastic and neoplastic epithelium of cervix uteri. However p16INK4a-negative CINs and carcinomas do exist. All stages of CINs and carcinomas analyzed are heterogeneous with respect to p16INK4a expression. So p16INK4a-negativity is not a sufficient reason to exclude a patient from the high risk group. As far as normal cervical epithelium is p16INK4a-negative and the ratio p16INK4a-positive/ p16INK4a-negative samples increases at the advanced stages application of immunohisto-/cytochemical test for p16INK4a may be regarded as a supplementary test for early diagnostics of cervical cancer. PMID- 15339340 TI - Hepatitis B virus variants in an HIV-HBV co-infected patient at different periods of antiretroviral treatment with and without lamivudine. AB - BACKGROUND: Lamivudine inhibits replication of both human immunodeficiency virus (HIV) and hepatitis B virus (HBV) and is commonly used as part of antiretroviral therapy. The main limitation in the use of lamivudine is resistant mutation selection. Most of these mutations affect the YMDD motif of the HBV DNA polymerase. The resistance occurs through M550V or M550I aminoacid replacements. The M550V variation may be accompanied by L526M mutation, notably in HIV-HBV co infected patients. The aim of this study was to investigate mutations associated with lamivudine resistance in a hemodialysis patient chronically co-infected with HIV-1 and HBV, who was submitted to several antiretroviral treatments. METHODS: HBV isolates derived from three blood samples collected at different times of antiretroviral therapies with and without lamivudine, were titred and submitted to nucleotide sequencing. RESULTS: HBV isolate derived from a sample collected in 1999 during an antiretroviral treatment with lamivudine showed the lamivudine resistant double mutation (L526M, M550V). However, no mutation associated with lamivudine resistance was observed in the HBV genome derived from the sample collected during a period of treatment without lamivudine (2001). After reinstitution of lamivudine (2002), the predominant HBV population exhibited a rare triple mutation (V519L, L526M, M550V), which has previously been associated with an in vitro reduction of virus antigenicity (escape mutant). HBV DNA was detected at high levels (108-109 copies/ml) in the three blood samples. CONCLUSIONS: Reintroduction of lamivudine as part of antiretroviral treatment in a patient who had developed lamivudine resistant HBV strains favored the predominance of an HBV isolate with reduced antigenicity. The absence of hepatitis acute exacerbation in this patient may be correlated to the absence of significant variations of the viral load, which was independent of the presence of mutations in the HBV DNA polymerase. PMID- 15339341 TI - A stochastic model for retinocollicular map development. AB - BACKGROUND: We examine results of gain-of-function experiments on retinocollicular maps in knock-in mice [Brown et al. (2000) Cell 102:77]. In wild type mice the temporal-nasal axis of retina is mapped to the rostral-caudal axis of superior colliculus. The established map is single-valued, which implies that each point in retina maps to a unique termination zone in superior colliculus. In homozygous Isl2/EphA3 knock-in mice the map is double-valued, which means that each point on retina maps to two termination zones in superior colliculus. This is because about 50 percent of cells in retina express Isl2, and two types of projections, wild-type and Isl2/EphA3 positive, form two branches of the map. In heterozygous Isl2/EphA3 knock-ins the map is intermediate between the homozygous and wild-type: it is single-valued in temporal and double-valued in the nasal parts of retina. In this study we address possible reasons for such a bifurcation of the map. RESULTS: We study the map formation using stochastic model based on Markov chains. In our model the map undergoes a series of reconstructions with probabilities dependent upon a set of chemical cues. Our model suggests that the map in heterozygotes is single-valued in temporal region of retina for two reasons. First, the inhomogeneous gradient of endogenous receptor in retina makes the impact of exogenous receptor less significant in temporal retina. Second, the gradient of ephrin in the corresponding region of superior colliculus is smaller, which reduces the chemical signal-to-noise ratio. We predict that if gradient of ephrin is reduced by a genetic manipulation, the single-valued region of the map should extend to a larger portion of temporal retina, i.e. the point of transition between single-and double-valued maps should move to a more nasal position in Isl2-EphA3 heterozygotes. CONCLUSIONS: We present a theoretical model for retinocollicular map development, which can account for intriguing behaviors observed in gain-of-function experiments by Brown et al., including bifurcation in heterozygous Isl2/EphA3 knock-ins. The model is based on known chemical labels, axonal repulsion/competition, and stochasticity. Possible mapping in Isl2/EphB knock-ins is also discussed. PMID- 15339342 TI - Dam inactivation in Neisseria meningitidis: prevalence among diverse hyperinvasive lineages. AB - BACKGROUND: DNA adenine methyltransferase (Dam) activity is absent in many, but not all, disease isolates of Neisseria meningitidis, as a consequence of the insertion of a restriction endonuclease-encoding gene, the 'dam replacing gene' (drg) at the dam locus. Here, we report the results of a survey to assess the prevalence of drg in a globally representative panel of disease-associated meningococci. RESULTS: Of the known meningococcal hyper-invasive lineages investigated, drg was absent in all representatives of the ST-8 and ST-11 clonal complexes tested, but uniformly present in the representatives of the other hyper invasive lineages present in the isolate collection (the ST-1, ST-4, ST-5, ST-32 and ST-41/44 clonal complexes). The patterns of sequence diversity observed in drg were consistent with acquisition of this gene from a source organism with a different G+C content, at some time prior to the emergence of present-day meningococcal clonal complexes, followed by spread through the meningococcal population by horizontal genetic exchange. During this spread a number of alleles have arisen by mutation and intragenic recombination. CONCLUSION: These findings are consistent with the idea that possession of the drg gene may contribute to the divergence observed among meningococcal clonal complexes, but does not have a direct mechanistic involvement in virulence. PMID- 15339343 TI - Most of the extant mtDNA boundaries in south and southwest Asia were likely shaped during the initial settlement of Eurasia by anatomically modern humans. AB - BACKGROUND: Recent advances in the understanding of the maternal and paternal heritage of south and southwest Asian populations have highlighted their role in the colonization of Eurasia by anatomically modern humans. Further understanding requires a deeper insight into the topology of the branches of the Indian mtDNA phylogenetic tree, which should be contextualized within the phylogeography of the neighboring regional mtDNA variation. Accordingly, we have analyzed mtDNA control and coding region variation in 796 Indian (including both tribal and caste populations from different parts of India) and 436 Iranian mtDNAs. The results were integrated and analyzed together with published data from South, Southeast Asia and West Eurasia. RESULTS: Four new Indian-specific haplogroup M sub-clades were defined. These, in combination with two previously described haplogroups, encompass approximately one third of the haplogroup M mtDNAs in India. Their phylogeography and spread among different linguistic phyla and social strata was investigated in detail. Furthermore, the analysis of the Iranian mtDNA pool revealed patterns of limited reciprocal gene flow between Iran and the Indian sub-continent and allowed the identification of different assemblies of shared mtDNA sub-clades. CONCLUSIONS: Since the initial peopling of South and West Asia by anatomically modern humans, when this region may well have provided the initial settlers who colonized much of the rest of Eurasia, the gene flow in and out of India of the maternally transmitted mtDNA has been surprisingly limited. Specifically, our analysis of the mtDNA haplogroups, which are shared between Indian and Iranian populations and exhibit coalescence ages corresponding to around the early Upper Paleolithic, indicates that they are present in India largely as Indian-specific sub-lineages. In contrast, other ancient Indian-specific variants of M and R are very rare outside the sub continent. PMID- 15339344 TI - Artificial neural network approach for selection of susceptible single nucleotide polymorphisms and construction of prediction model on childhood allergic asthma. AB - BACKGROUND: Screening of various gene markers such as single nucleotide polymorphism (SNP) and correlation between these markers and development of multifactorial disease have previously been studied. Here, we propose a susceptible marker-selectable artificial neural network (ANN) for predicting development of allergic disease. RESULTS: To predict development of childhood allergic asthma (CAA) and select susceptible SNPs, we used an ANN with a parameter decreasing method (PDM) to analyze 25 SNPs of 17 genes in 344 Japanese people, and select 10 susceptible SNPs of CAA. The accuracy of the ANN model with 10 SNPs was 97.7% for learning data and 74.4% for evaluation data. Important combinations were determined by effective combination value (ECV) defined in the present paper. Effective 2-SNP or 3-SNP combinations were found to be concentrated among the 10 selected SNPs. CONCLUSION: ANN can reliably select SNP combinations that are associated with CAA. Thus, the ANN can be used to characterize development of complex diseases caused by multiple factors. This is the first report of automatic selection of SNPs related to development of multifactorial disease from SNP data of more than 300 patients. PMID- 15339345 TI - Bayesian model accounting for within-class biological variability in Serial Analysis of Gene Expression (SAGE). AB - BACKGROUND: An important challenge for transcript counting methods such as Serial Analysis of Gene Expression (SAGE), "Digital Northern" or Massively Parallel Signature Sequencing (MPSS), is to carry out statistical analyses that account for the within-class variability, i.e., variability due to the intrinsic biological differences among sampled individuals of the same class, and not only variability due to technical sampling error. RESULTS: We introduce a Bayesian model that accounts for the within-class variability by means of mixture distribution. We show that the previously available approaches of aggregation in pools ("pseudo-libraries") and the Beta-Binomial model, are particular cases of the mixture model. We illustrate our method with a brain tumor vs. normal comparison using SAGE data from public databases. We show examples of tags regarded as differentially expressed with high significance if the within-class variability is ignored, but clearly not so significant if one accounts for it. CONCLUSION: Using available information about biological replicates, one can transform a list of candidate transcripts showing differential expression to a more reliable one. Our method is freely available, under GPL/GNU copyleft, through a user friendly web-based on-line tool or as R language scripts at supplemental web-site. PMID- 15339346 TI - Estimating mutual information using B-spline functions--an improved similarity measure for analysing gene expression data. AB - BACKGROUND: The information theoretic concept of mutual information provides a general framework to evaluate dependencies between variables. In the context of the clustering of genes with similar patterns of expression it has been suggested as a general quantity of similarity to extend commonly used linear measures. Since mutual information is defined in terms of discrete variables, its application to continuous data requires the use of binning procedures, which can lead to significant numerical errors for datasets of small or moderate size. RESULTS: In this work, we propose a method for the numerical estimation of mutual information from continuous data. We investigate the characteristic properties arising from the application of our algorithm and show that our approach outperforms commonly used algorithms: The significance, as a measure of the power of distinction from random correlation, is significantly increased. This concept is subsequently illustrated on two large-scale gene expression datasets and the results are compared to those obtained using other similarity measures.A C++ source code of our algorithm is available for non-commercial use from kloska@scienion.de upon request. CONCLUSION: The utilisation of mutual information as similarity measure enables the detection of non-linear correlations in gene expression datasets. Frequently applied linear correlation measures, which are often used on an ad-hoc basis without further justification, are thereby extended. PMID- 15339347 TI - Could a simple surgical intervention eliminate HIV infection? AB - BACKGROUND: Human Immunodeficiency Virus (HIV) infection is a dynamic interaction of the pathogen and the host uniquely defined by the preference of the pathogen for a major component of the immune defense of the host. Simple mathematical models of these interactions show that one of the possible outcomes is a chronic infection and much of the modelling work has focused on this state. BIFURCATION: However, the models also predict the existence of a virus-free equilibrium. Which one of the equilibrium states the system selects depends on its parameters. One of these is the net extinction rate of the preferred HIV target, the CD4+ lymphocyte. The theory predicts, somewhat counterintuitively, that above a critical extinction rate, the host could eliminate the virus. The question then is how to increase the extinction rate of lymphocytes over a period of several weeks to several months without affecting other parameters of the system. TESTING THE HYPOTHESIS: Proposed here is the use of drainage, or filtration, of the thoracic duct lymph, a well-established surgical technique developed as an alternative for drug immunosuppression for organ transplantation. The performance of clinically tested thoracic duct lymphocyte depletion schemes matches theoretically predicted requirements for HIV elimination. PMID- 15339361 TI - Training in genitourinary medicine from the specialist registrar's point of view: opportunities and facilities available across the UK and plans for future working. AB - In recent years training for specialist registrars (SpRs) in GU medicine has undergone a number of changes. In addition, the speciality in general is undergoing modernization and change and issues of workforce planning and consultant job availability have been of particular importance to SpRs. In March 2003 a postal survey of SpRs in the UK was undertaken to evaluate training, future career plans and working patterns. A 59% (69/117) response rate was achieved and overall most SpRs expressed satisfaction with their training. However, a number of concerns were raised with aspects of training, which are discussed in this paper. Nearly half of SpRs will consider working part time in the future and almost two-thirds are interested in job sharing. This is the first survey of its kind in GU medicine and the results are informative to all involved with SpR training programmes and workforce planning in the speciality. PMID- 15339362 TI - Immunology of HIV--filling in the details. PMID- 15339363 TI - How do clients access their HIV test results from genitourinary medicine services in the UK? A time for change! AB - With the changes in the nation's sexual health needs and the introduction of the Government's Sexual Health Strategy, GU services need to review their operational policies to accommodate the inevitable increase for service. There had been a long-standing practice of anyone having an HIV test to return in person to collect his or her result. As more and more people, many of whom are considered to be low risk, are testing for HIV as part of a routine sexual health check-up, the question is, do they really need to return in person for these results? The national survey shows that while the majority of clinics are insisting that everyone having an HIV test returns in person, there are some services that have already adopted other approaches. It is also evident that many service providers are also considering a change in policy. PMID- 15339364 TI - Understanding access to genitourinary medicine services. AB - In this study we describe waiting times in genitourinary medicine clinics (England, Wales, Northern Ireland), describe patterns of clinic attendance over time, and explore the association between waiting times and attendance patterns. A postal survey of clinicians in October 2002 explored waiting times and appointment policy. Clinic attendance data were linked to survey responses, and attendance trends described in relation to national sexually transmitted infection (STI) data. Waiting times were reported, and associations between attendance patterns and appointment policy explored. Mean waiting times were 11.2 days for males and 12.5 days for females. An association between longer waiting time and new males' non-attendance rates was shown. From 1996 to 2002 there was no overall increase in attendances in English clinics, while the proportion of all patients who were new or had a new problem increased from 0.37 to 0.44, and the ratio of new STI diagnoses to new attenders increased from 0.2 to 0.31. Major acute STIs diagnosed in England increased from 114,380 to 185,247 cases. PMID- 15339365 TI - Sexual behaviour among young men in Sweden and the impact of pornography. AB - The purpose was to investigate the sexual behaviour among young men (n = 300), visiting a genitourinary clinic in Sweden, focusing on the impact of pornography. Almost all, 98% (n = 292) claimed to be heterosexual. The mean age at first intercourse was 16 years and on that occasion 64% (n = 187) used some kind of contraceptive, mainly condom. All, 99% (n = 296) had consumed pornography and 53% (n = 157) felt that pornography impacted their sexual behaviour; they got inspired. About half (n = 161) had had anal intercourse. Of these, 70% (n = 113) had had it more than once and 84% (n = 133) could imagine doing it again. Only 17% (n = 28) always used a condom in this situation. One out of four (n = 70) had had at least one sexually transmitted disease. The low use of condoms when heterosexual men have anal sex might have serious consequences for a spread of sexually transmitted diseases. PMID- 15339366 TI - HIV infection in the era of highly active antiretroviral therapy: the Malaga Study. AB - We analyse the characteristics of patients diagnosed with HIV infection in the highly active antiretroviral therapy era in the southeast of Spain. Data were collected on 470 HIV patients diagnosed between January 1997 and December 2002. The number of cases fell over recent years and HIV transmission was sexual in 70.5%. The mean CD4 lymphocyte count was 302.1 x 10(6)/L and the mean viral load 4.70 log(10). Diagnosis of HIV coincided with an AIDS-defining opportunistic illness in 30.6% of patients and a late diagnosis (CD4 < 200 x 10(6)/L) was made in 48.3% of patients. A late diagnosis was related to male gender (OR 2.50; 95% CI 1.20-5.12; P < 0.001) and AIDS case (OR 18.80; 95% CI 10.50-33.80; P < 0.00001). These results suggest that there has been a progressive reduction in new cases of HIV-infected patients, with the main route of transmission being sexual and that the diagnosis was late in almost half the patients. PMID- 15339367 TI - Herpes simplex virus infection of the central nervous system in human immunodeficiency virus-type 1-infected patients. AB - We report clinical, radiological and virological data from nine consecutive HIV infected patients with herpes simplex virus (HSV) infection of the central nervous system (CNS). Three patients presented with confusion, two with fever and headache, two with anxiety and depression, one with slow mentation and memory loss and one with expressive dysphasia. Five patients had previous AIDS-defining diagnoses: four of these five patients had previous cutaneous HSV infection. HSV DNA was detected by the polymerase chain reaction (PCR) on cerebrospinal fluid (CSF) in seven patients. HSV infection was diagnosed by brain biopsy (after negative PCR on CSF) in one patient and at autopsy in one patient (after negative CSF PCR and brain biopsy). Seven patients received specific anti-viral therapy; two died of unrelated causes and the other five recovered. Two patients were not treated, in one the diagnosis was made at autopsy and the other recovered spontaneously. HIV-infected patients with CNS HSV infection have a varied presentation. Diagnosis by PCR on CSF identified the majority of cases. With specific treatment the outcome was good. PMID- 15339368 TI - High-risk sexual behaviours among drug users in Pakistan: implications for prevention of STDs and HIV/AIDS. AB - Our objective was to describe HIV/STD risk behaviours and awareness among a community-based sample of drug users in Pakistan. Drug users contacted through street outreach by a non-governmental organization in Quetta, Peshawar and Rawalpindi underwent interviewer-administered questionnaires. Descriptive statistics were used to characterize sexual behaviours by city, marital status and the use of injection drugs. Logistic regression was used to identify correlates of ever having an STD. Of 608 drug users studied, all but one was male; median age was 32 years and 45% had no formal education. Half were married, of whom 25% were living with their wives. Sexual behaviours were reported as follows: 14% had sex with other males, 28% reported sex with both males and females, 49% had paid money to have sex and only 10% had ever used condoms. One fifth reported having had an STD and about 40% reported having suffered from either one or more STD-related symptoms. Only 41% had heard about HIV/AIDS, of whom 17% knew that HIV/AIDS could be transmitted through sexual contact. In conclusion, high-risk sexual behaviours are prevalent among male drug users in Pakistan, and awareness of transmission risks is low. These data attest to the urgent need for effective and specific interventions in Pakistan to prevent transmission of HIV and STDs among drug users and their sex partners. PMID- 15339369 TI - Determinants of HIV antibody testing among selected groups of Chinese residents in Hong Kong. AB - HIV testing is promoted to the public as a tool for HIV prevention in many countries. However, the patterns and determinants of HIV antibody testing among the Chinese people are unknown. To describe the prevalence and determinants of HIV antibody testing amongst the Hong Kong Chinese a cross-sectional anonymous survey was carried out among 1,027 subjects from different population groups (airport travellers, business sector workers, service sector workers, university staff, and STD clinic attendees). Subjects were categorized into either 'STD population' (respondents from STD clinics) versus 'all others' (respondents from other settings) groups. Forty-five percent of the respondents reported ever having had a test for HIV antibody. 'STD population' group members were almost three times more likely to have had an HIV test than were 'all others' group members (77% vs 20%). After adjustments, 'STD population' group members who reported having tested for HIV were more often aged 45 years or older, alcohol drinkers, with high self-efficacy scores, and inconsistent condom users; members in the 'all others' group, more often had had sex with strangers, drank alcohol, and favoured having multiple sex partners. Awareness among the public about the risk behaviours for HIV should be enhanced and efforts should be made to reduce high-risk behaviours among those tested by emphasizing the importance of maintaining safer sex behaviour and having follow-up tests during post-test counselling. PMID- 15339370 TI - Association of Trichomonas vaginalis with sociodemographic factors and other STDs among female inmates in Lisbon. AB - A study on STDs was conducted among 211 female inmates in a prison in Lisbon, Portugal, in order to establish possible associations between Trichomonas vaginalis infection, sociodemographic factors and other STDs. T. vaginalis was found in 31.2% of the women, from whom only 65.1% presented symptoms. It was more frequently isolated in the 20-40 year age group. On univariate analysis there was an association, although not statistically significant, between the existence of T. vaginalis, multiple sexual partners, drug addiction and no condom use. A statistically significant association was found between trichomoniasis, prostitution and other STDs. These were found in 87% of all women. T. vaginalis was detected alone in 23.8% (15/63) of all women with trichomoniasis, while 76.2% (48/63) of them had multiple infections with trichomonas and other STDs. A statistically significant association was present between trichomoniasis and Mycoplasma hominis and infection with Treponema pallidum. This study showed that the prevalence of T. vaginalis is as high as that of multiple infections with other STDs. Therefore, our findings seem to confirm that trichomoniasis serves as a marker for other STDs. Screening for STDs should then be offered to Portuguese inmates, or, at least, these women should be screened for T. vaginalis, as a marker for the other STDs. PMID- 15339371 TI - MOTION: Experts have not just been 'seeing what they wanted to see' by 'ignoring' health care transmission of AIDS in Africa. Sexual transmission is indeed the major mode of transmission. PROPOSAL: Initiatives to prevent sexual transmission of HIV in Africa should not be over-shadowed by current debate. PMID- 15339373 TI - Improving uptake of HIV testing in patients with a confirmed STI. AB - Up to 70% of GU clinic attendees with a sexually transmitted infection (STI) and undiagnosed HIV, remain HIV undiagnosed after their visit. STIs have been shown to facilitate HIV transmission. Therefore, patients with an STI should test for HIV. Our objective was to compare the uptake of HIV testing in GU clinic attendees with an STI (study group) with those receiving a negative GU screen (control group). This re-audit was performed after introduction of the following clinic changes: nurse-performed asymptomatic GU screening; 'opt-out' HIV testing policy; discontinuing routine pre-HIV test counselling; access to HIV results by post. The uptake of HIV testing overall, and in the study and control groups respectively was 53% (n = 573), 41% (n = 285) and 65%, showing significant improvement compared to the first audit in 1999 (18%, 14% and 33% respectively) (P < 0.01). The clinic interventions increased HIV testing rates which were in keeping with National Sexual Health strategy targets. PMID- 15339374 TI - HIV partner notification: a 2002 Victorian audit. AB - This audit was carried out in August 2003 and examined HIV notification data in Victoria. We aimed to determine the proportion of partners reported as being traced from newly diagnosed HIV individuals, the type of contact tracing used and identify the number of HIV cases newly diagnosed from contact tracing. We compared men who have sex with men (MSM) with all other newly diagnosed HIV individuals in Victoria (non-MSM). Of the 215 newly diagnosed HIV-positive individuals, 49% (n = 105) could potentially carry out contact tracing. There was a difference between MSM and non-MSM in the proportion of contactable partners (44% vs 63% respectively, P = 0.017). MSM less commonly used provider referral than non-MSM (12% vs 53% respectively, P < 0.001) but more commonly used patient referral (41% MSM vs 9% non-MSM, P = 0.001). Of the 13 individuals found to be HIV-positive through contact tracing, nine (69%) were non-MSM. PMID- 15339375 TI - Osteomyelitis complicating pyomyositis in HIV disease. AB - Pyomyositis has previously been described in association with human immunodeficiency virus (HIV) and as a discrete entity in HIV seronegative patients from tropical climates (tropical pyomyositis). Pyomyositis and osteomyelitis are usually considered a late complication of advanced HIV disease. We describe a patient with well-controlled HIV and both types of musculoskeletal infection. The case highlights an unusual presentation, the utility of MRI in soft tissue infection and an excellent outcome from prolonged antimicrobial therapy following surgical debridement. PMID- 15339376 TI - Contraceptive practices amongst female genitourinary medicine clinic attenders. PMID- 15339377 TI - Inconsistencies in partner notification practices in genitourinary medicine clinics--time to put our house in order. PMID- 15339378 TI - Can a shrinking service mislead people into thinking there is less disease? PMID- 15339379 TI - General practitioners and Sexual Health Strategy on Sexual Health and HIV. PMID- 15339380 TI - Are we really assessing HIV/AIDS-related knowledge? Competing beliefs that may put a 'knowledgeable' person at risk for HIV. PMID- 15339381 TI - Treatment of erectile dysfunction. PMID- 15339382 TI - Gonorrhoea: test of cure? PMID- 15339383 TI - Involvement of cyclin dependent kinase 5 and its activator p35 in staurosporine induced apoptosis of cortical neurons. AB - AIM: To investigate whether cyclin-dependent kinase 5 and its regulatory protein p35 was involved in staurosporine-induced apoptosis of cortical neuronal cultures. METHODS: Primary cerebral cortical neurons were exposed to 300 nmol/L staurosporine. After incubation for different time, morphological alterations were observed with phase-contrast microscopy, fluorescence microscopy, and transmission electron microscopy. DNA fragmentation was detected by agarose gel electrophoresis. The protein levels of Cdk4, p53, Cdk5, and its regulatory protein p35 following staurosporine treatment were measured by Western blotting. The Cdk5 activity was assayed for histone H1 kinase activity by autoradiography. RESULTS: The typical morphological changes of apoptosis were observed and the nuclear DNA fragmentation showed the characteristic "ladder" pattern after the cells were treated by staurosporine. The Cdk5 protein level increased markedly at 3 h and continued to 24 h. The p35 level increased at 3 h after being exposed to staurosporine, and decreased at 12 h. The cleavage of p35 to p25 was also detected at 12 h and increased at 24 h. There was no increase in Cdk5 kinase activity despite the increased cleavage of p35. The protein level of Cdk4 protein increased at 3 h and then decreased gradually from 6 h, but it was still higher than that in the vehicle cultures at 12 h. The p53 level decreased obviously at 3 h after staurosporine treatment and then seemed to increase at 12 h, but remained lower than that of vehicle cultures. CONCLUSION: Staurosporine-induced increase in Cdk5 protein levels and the cleavage of p35 to p25 may contribute to neuronal apoptosis. PMID- 15339384 TI - N-Acetyltransferase 2 gene polymorphism in a group of senile dementia patients in Shanghai suburb. AB - AIM: To investigate the possible association of hereditary polymorphism of N acetyltransferase 2 (NAT2) gene with the susceptibility towards senile dementia in farmer population of Shanghai suburb. METHODS: NAT2 gene genotyping was performed at 7 major polymorphic loci (G191A, C282T, T341C, C481T, G590A, A803G, and G857A) with a polymerase chain reaction-based restriction fragment length polymorphism based procedure in 2 groups of farmer subjects in Shanghai suburb. A group of 51 diagnosed dementia patients [comprising 29 sporadic Alzheimer disease (AD) patients and 22 sporadic vascular dementia (VD) patients] and a group of 112 healthy individuals were in the same area. RESULTS: The homogenous rapid genotypes (R/R, including*4/*4, *13 /*13, and *4/*13) was found over-present in both groups of patients, compared with healthy individuals, for all farmer dementia patients, 52.9 % vs 33.0 %, P=0.016, OR (95 % CI): 2.28(1.16-4.48); for AD group only, 51.7 % vs 33.0 %, P=0.063, OR (95 % CI): 2.18 (0.95-4.97); for VD group 54.5 % vs 33.0 %, P=0.055, OR (95 % CI): 2.43 (0.96-2.43). The significant frequency difference of genotype *4/*7B between farmer dementia patients and healthy individuals, and that of solo-alleles *13, and *7B were observed between the healthy individuals and both groups of dementia patients. CONCLUSION: Our data suggest the involvement of various NAT2 rapid-acetylating genotypes in the individual susceptibility to senile dementia. Variant genotypes of NAT2 might serve as a hereditary risk factor for AD and VD in Chinese population. PMID- 15339385 TI - Effect of combination of extracts of ginseng and ginkgo biloba on acetylcholine in amyloid beta-protein-treated rats determined by an improved HPLC. AB - AIM: To determine the concentration of acetylcholine (ACh) in amyloid beta protein (Abeta) treated rats and offer a method determining ACh as well. METHODS: A 1-month combination of extrats of ginseng and ginkgo biloba (Naoweikang) ig administration to rats was performed daily after bilateral injection of Abeta(1 40) (4 g/L, 1 microL for each side) into hippocampus. After decollation, homogenizing, and centrifuging and extracting, a high pressure liquid chromatographic (HPLC) method using electrochemical detection (ECD) combined with two immobilized enzyme reactors was used to determine ACh in rat whole brain. RESULTS: With a mobile phase consisting of disodium hydrogen orthophosphate, tetramethylammonium chloride (TMACl), octanesulfonic acid sodium salt (OSA) and "Reagent MB" at a final pH of 8.0, ACh was determined while removing the interfering choline in less than 10 min at a flow rate of 0.35 mL/min on a platinum (Pt) working electrode at a potential of +300 mV vs a solid-state palladium (Pd) reference electrode. Linear regression analysis of peak area vs concentration demonstrated linearity in the 28.01 to 1400.06 microg/L injection range. The r-value was 0.9978. The limit of detection (LOD) is 0.28 ng on column. ACh in whole brain decreased by 20.34 % (from 162.1+/-32.7 to 134.7+/-14.0 microg/L, P<0.05) after bilateral injection of Abeta into rat hippocampus. After Naoweikang administration (31 and 15.5 mg/kg, respectively), ACh increased by 19.97 % (from 134.7+/-14.0 to 161.6+/-26.2 microg/L, P<0.05) and 18.56 % (from 134.7+/-14.0 to 159.7+/-22.9 microg/L, P<0.05), respectively. CONCLUSION: Naoweikang significantly increased the level of ACh in whole brain of Abeta treated rats. And a sensitive, selective and reliable method for routinely determining ACh in rat whole brain was established in this study. PMID- 15339386 TI - Proteomic response analysis of endothelial cells of human coronary artery to stimulation with carbachol. AB - AIM: To identify the molecular basis of the endothelial target for acetylcholine (ETA). METHODS: Proteomic methods were used to monitor changes in protein expression in the first 10 h following the stimulation of human coronary endothelial cells with carbachol 100 micromol/L. Thirty proteins showing the largest changes were identified by mass spectrometry. RESULTS: Based on analysis with Image Master 2-D Elite software, about 623 protein spots were detected in control cells and 825 protein spots in carbachol-treated cells, the matching rate was 68.1 %. Among all the detected spots, 39 were up-regulated and 29 were down regulated, showing detectable changes varied from 1.7-3.8 folds. Forty one spots in the peptide mass fingerprints were successfully obtained. The most interesting feature was that all the four newly synthesized proteins belonged to the heat shock protein family. CONCLUSION: These identified proteins played key roles in the molecular mechanism of ETA. PMID- 15339387 TI - Ghrelin protects myocardium from isoproterenol-induced injury in rats. AB - AIM: To investigate the cardiac protective effects of ghrelin in rat with myocardial injury induced by isoproterenol (ISO). METHODS: Rats were subcutaneously injected ISO 40 mg/kg/d with or without ghrelin 1 or 10 nmol/kg/d for 2 d. Hemodynamic parameters including mean arterial blood pressure and left ventricular pressure were measured at 12 h after the last injection with ISO and/or ghrelin. Plasma lactate dehydrogenase (LDH) activity, plasma and myocardial contents of malondialdehyde (MDA), and conjugated diene were measured. Plasma ghrelin and endothelin-1 levels were assayed using radioimmunoassay methods. Endothelin-1 and ghrelin mRNA were determined using RT-PCR. RESULTS: About 45 % (5/11) of rats after treatment with ISO alone died during experimental periods. However, no rats died after administration with ghrelin 10 nmol/kg/d (0/11, P<0.05). Ghrelin also obviously ameliorated the hemodynamic disturbance in rats induced by ISO. The plasma LDH activity, contents of myocardial and plasma MDA, and conjugated diene level in plasma in ISO+G10 nmol/kg/d group were decreased by 28 %, 34 %, 73 %, and 38 % compared with those of ISO group (all P<0.01) respectively. ISO-induced endothelin-1 mRNA over-expression was inhibited and endothelin-1 level in plasma were inhibited by ghrelin 1 and 10 nmol/kg/d. The ghrelin levels in plasma and ghrelin mRNA in myocardium were increased in the rats after injection of ISO. The plasma ghrelin level was further increased after ghrelin administration. CONCLUSION: Ghrelin has a protective effect against ISO induced myocardial injury. PMID- 15339388 TI - Tetrahydroacridine inhibits voltage-dependent Na(+) current in guinea-pig ventricular myocytes. AB - AIM: To study the effects of tetrahydroacridine (tacrine) on voltage-gated Na(+) channels in cardiac tissues. METHODS: Single ventricular myocytes were enzymatically dissociated from adult guinea-pig heart. Voltage-dependent Na(+) current was recorded using whole cell voltage-clamp technique. RESULTS: (1) Tacrine reversibly inhibited Na(+) current with an IC(50) value of 120 micromol/L (95 % confidence range: 108-133 micromol/L). (2) The inhibitory effects of tacrine on Na(+) current exhibited both a tonic nature and use-dependence. (3) Tacrine at 100 micromol/L caused a negative shift (about 10 mV) in the voltage dependence of steady-state inactivation of Na(+) current, and retarded its recovery from inactivation, but did not affect its activation curve. (4) Intracellular application of tacrine significantly inhibited Na(+) current. CONCLUSION: In addition to blocking other voltage-gated ion channels, tacrine blocked Na(+) channels in guinea-pig ventricular myocytes. Tacrine acted as inactivation stabilizer of Na(+) channels in cardiac tissues. PMID- 15339389 TI - Effect of angiotensin II type 1 receptor on delayed rectifier potassium current in catecholaminergic CATH.a cells. AB - AIM: To study the modulatory effects of angiotensin II (Ang II) on the delayed rectifier potassium (Kv) current (IKv) and its underlying intracellular mechanism in the catecholaminergic system of rats. METHODS: AT1 and AT2 receptors of the differentiated and undifferentiated CATH.a cells were determined by radioligands binding assay. The IKv was recorded with the whole cell patch-clamp configuration in voltage clamp mode on CATH.a cells. RESULTS: The Ang II receptor proteins including AT1 and AT2 receptors were expressed in CATH.a cells, and the number of the former was significantly more than the latter (P<0.01). The IKv of CATH.a cells was reduced by superfusion with the Ang II (100 nmol/L) (P<0.05) in the presence of the AT2 receptor antagonist PD123319, but was not affected by only superfusion with PD123319. The effect of Ang II on IKv in CATH.a cells was completely inhibited by addition of AT1 receptor antagonist losartan. Superfusion with Ang II (100 nmol/L) plus U73122, an inhibitor of phospholipase C (PLC) in the presence of PD123319 had no effect on the IKv [(20.2+/-2.8) pA/pF]. Ang II induced reduction of IKv was attenuated (P<0.05) but not abolished by PKC inhibitor calphostin C (Cal) and selective CaMK II inhibitor KN-93 (10 micromol/L) respectively. However, IKv reduction was completely abolished by superfusion with both Cal and KN-93. CONCLUSION: The inhibition of Kv currents in CATH.a cells by Ang II is mediated by AT1 receptor, and the PLC, PKC and CaMK II may be involved in signal transduction of AT1 receptor. The differentiated CATH.a cell is a useful cell model to study Ang II receptor-mediated functional modulation of catecholaminergic system. PMID- 15339390 TI - Total saponins of Panax notoginseng protected rabbit iliac artery against balloon endothelial denudation injury. AB - AIM: To investigate whether total Panax notoginseng saponins (PNS) could protect endothelium of rabbit iliac artery against balloon endothelial denudation (BED) injury. METHODS: The morphology changes of the endothelium were observed with scanning electron microscope (SEM) and hematoxylin and eosin stain after BED of rabbit iliac artery at 0, 4, 6, and 8 week respectively. Vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) was also determined by immunohistochemistry. PNS 10, 30, and 50 mg/kg were administered iv per day from 2 d before to 4 weeks after operation. RESULTS: The endothelium was denudated completely after BED. At the 4th week the endothelium was repaired in some degree, then recovered gradually at 6 and 8 week. The degree of intimal thickening at 4 week was significantly greater than that at 0, 6, or 8 week. The sequence of VEGF or MMP-2 staining from strong to weak was 4, 6, 0, 8 week, and normal control. However at 4 week, endothelial regeneration in PNS 30 and 50 mg/kg groups was significantly faster than that in saline group. The intimal thickness was significantly decreased and expressions of VEGF and MMP-2 were both down-regulated in PNS 30 or 50 mg/kg groups compared with saline control group. CONCLUSION: PNS promoted the endothelial regeneration and reduced ECM thickening, which was related to regulation of the expression of VEGF and MMP-2. PNS may have sustained antirestenotic effect after BED. PMID- 15339391 TI - Different contributions of STAT3, ERK1/2, and PI3-K signaling to cardiomyocyte hypertrophy by cardiotrophin-1. AB - AIM: To assess the contribution of signal transducer and activator of transcription 3 (JAK-STAT3) pathway, extracellular signal-regulated kinases1/2 (ERK1/2) pathway, and phosphatidylinositol 3-kinase (PI3-K) pathway to cardiomyocytes hypertrophy induced by cardiotrophin-1 (CT-1), a new member of interleukin-6 (IL-6) family of cytokines. METHODS: STAT3, ERK1/2, and PI3-K were assessed by Western blot analysis. Activity of ERK1/2 was also confirmed by in gel kinase assay. Hypertrophy of cardiomyocyte was evaluated by [3H]leucine incorporation and cellular protein-to-DNA ratio. RESULTS: CT-1 simultaneously activated phosphorylation of STAT3, ERK1/2, and PI3-K in rat cardiomyocytes. Parthenolide, an inhibitor of STAT, suppressed CT-1-induced [3H]leucine incorporation by 88.3 % and protein-to-DNA ratio by 75.0 %. U0126, an MEK1/2 inhibitor, increased CT-1-induced the phosphorylation of STAT3 in a dose dependent manner and, consistently, augmented CT-1-induced increase in [3H]leucine incorporation and cellular protein-to-DNA ratio by 17.6 % and 16.3 %, respectively. Wortmannin, a PI3-K inhibitor, did not influence CT-1-induced [3H]leucine incorporation and cellular protein-to-DNA ratio. CONCLUSION: The hypertrophic effect of CT-1 was essentially mediated by STAT3, independent of PI3 K, and negatively regulated by ERK1/2 via inhibiting the phosphorylation of STAT3. The interaction between STAT3 and ERK1/2 in CT-1-induced signaling contributes to development of cardiac hypertrophy. PMID- 15339392 TI - Establishment of a cell-based assay to screen regulators for Klotho gene promoter. AB - AIM: To discover compounds which can regulate Klotho promoter activity. Klotho is an aging suppressor gene. A defect in Klotho gene expression in the mouse results in the phenotype similar to human aging. Recombinant Klotho protein improves age associated diseases in animal models. It has been proposed that up-regulation of Klotho gene expression may have anti-aging effects. METHODS: Klotho promoter was cloned into a vector containing luciferase gene, and the reporter gene vector was transfected into HEK293 cells to make a stable cell line (HEK293/KL). A model for cellular aging was established by treating HEK293/KL cells with H2O2. These cells were treated with extracts from Traditional Chinese Medicines (TCMs). The luciferase activity was detected to identify compounds that can regulate Klotho promoter. RESULTS: The expression of luciferase in these cells was under control of Klotho promoter and down-regulated after H2O2 treatment. The down-regulation of luciferase expression was H2O2 concentration-dependent with an IC50 at approximately 0.006 %. This result demonstrated that the Klotho gene promoter was regulated by oxidative stress. Using the cell-based reporter gene assay, we screened natural product extracts for regulation of Klotho gene promoter. Several extracts were identified that could rescue the H2O2 effects and up-regulated Klotho promoter activity. CONCLUSION: A cell -based assay for high-throughput drug screening was established to identify compounds that regulate Klotho promoter activity, and several hits were discovered from natural products. Further characterization of these active extracts could help to investigate Klotho function and aging mechanisms. PMID- 15339393 TI - Selective inhibition of purified human phosphodiesterase 4A expressed in yeast cell GL62 by ciclamilast, piclamilast, and rolipram. AB - AIM: To improve the specific activity of human phosphodiesterase 4A (PDE4A) expressed in yeast cell GL62 and investigate the effects of selective phosphodiesterase 4 (PDE4) inhibitors (ciclamilast, piclamilast, and rolipram), selective phosphodiesterase 5 (PDE5) inhibitor zaprinast, and cyclooxygenase (COX) inhibitors (aspirin, indomethacin) on human PDE4A activity expressed in yeast cell GL62. METHODS: Human PDE4A was expressed in yeast cell GL62 after CuSO4 induction and the specific activity of human PDE4A was improved by ammonium sulfate fractionation, DEAE Sephadex A-50 chromatography, and Sephadex G-100 chromatography. The activity of PDE4A was measured by high performance liquid chromatography (HPLC). RESULTS: Induced PDE4A activity expressed in crude yeast cell GL62 supernatant and pellet was (340+/-21) nmol/g/min and (250+/-25) nmol/g/min respectively. The specific activity of recombinant PDE4A in supernatant was improved 6.4 fold. Ciclamilast, piclamilast, and rolipram could inhibit PDE4A activity. The IC50 values (95 % confidence limits) of ciclamilast, piclamilast, and rolipram were 1.27 (0.84-1.91), 66.4 (33.3-132.2), and 3.73 (2.51-5.53) micromol/L respectively. Zaprinast, aspirin, and indomethacin had no obvious inhibitory effect on PDE4A activity. CONCLUSION: The specific activity of PDE4A expressed in yeast cell GL62 can be improved by ammonium sulfate fractionation, DEAE Sephadex A-50 chromatography, and Sephadex G-100 chromatography. Ciclamilast, piclamilast, and rolipram can inhibit PDE4A activity while zaprinast, aspirin, and indomethacin have no obvious inhibitory effect on PDE4A activity. Human PDE4A expressed in GL62 might be useful in the research and screening of new selective PDE4 inhibitors. PMID- 15339394 TI - Induction of mucin secretion from human bronchial tissue and epithelial cells by rhinovirus and lipopolysaccharide. AB - AIM: To examine the effects of rhinovirus and lipopolysaccharide (LPS) on mucin secretion from bronchial tissue and epithelial cells in vitro. METHODS: Human small bronchial tissue fragments (HSBTF) and human bronchial epithelial cells (HBEC) were cultured with rhinovirus 16 and LPS, respectively and culture supernatants were collected for mucin measurement. To determine mucin levels in the culture supernatants, a MUC5AC enzyme linked immunosorbent assay and an enzyme linked lectin assay procedure with dolichos biflorus agglutinin (DBA) were developed, and mucin release was expressed as percentage increased (or decreased) secretion over baseline level. RESULTS: A concentration-dependent release of DBA mucin and MUC5AC mucin were observed when HSBTF were infected with various concentrations of rhinovirus 16 at 37 degree. The maximum-induced DBA mucin and MUC5AC mucin release were approximately 258 % and 83 % over baseline. The response of HSBTF to rhinovirus was completely abolished by metabolic inhibitors. Rhinovirus was also able to induce a concentration-dependent release of DBA mucin and MUC5AC mucin from primarily cultured HBEC. LPS 100 mg/L was able to provoke up to approximately 19 % and 54 % increase in DBA and MUC5AC mucin release over baseline, respectively from HSBTF, and 3.1 % and 57 % increase from HBEC at 20 h. Soybean trypsin inhibitor (SBTI) 30 mg/L was able to inhibit LPS-induced mucin release from HSBTF and HBEC. CONCLUSION: Rhinovirus is able to induce mucin secretion from human bronchial tissue and bronchial epithelial cells in vitro. LPS can induce MUC5AC mucin release from HSBTF and HBEC. PMID- 15339395 TI - Anti-adjuvant arthritis of recombinant human endostatin in rats via inhibition of angiogenesis and proinflammatory factors. AB - AIM: To investigate the profile of endostatin on adjuvant arthritis (AA) and angiogenesis blockade in synovitis. METHODS: The model of rat AA was induced by injection of intradermal complete Freund's adjuvant (CFA). Hind paw volume of rat was measured by volume meter and the activities of interleukin-1 (IL-1) and IL-2 were measured by the assay of thymocytes proliferation. IL-1beta and tumor necrosis factor-alpha (TNF-alpha) produced by synoviocytes was estimated with radioimmunoassay. The number of new blood vessels in knee joint synovium was counted under microscope by hematoxylin and eosin (HE) staining. RESULTS: The secondary inflammation of AA rats appeared on the 10th day after injection of CFA. The therapeutic administration of endostatin (0.1, 0.5, and 2.5 mg/kg/d, sc, plus 7 d) was given from that time (d 10). It was found that endostatin significantly inhibited the secondary paw swelling and the number of new blood vessels in the synovium of AA rats. Endostatin significantly decreased the production of IL-1 derived from both peritoneal macrophages and synoviocytes and IL-2 from splenocytes, especially at the dose of 2.5 mg/kg. This effect of endostatin also was seen on TNF-alpha produced by synoviocytes. CONCLUSION: The recombinant human endostatin had an inhibitory effect on rat AA, which was related to its anti-angiogenesis and inhibition of proinflammatory cytokines. PMID- 15339396 TI - Learning of medical pharmacology via innovation: a personal experience at McMaster and in Asia. AB - Pharmacology, a discrete preclinical discipline of the traditional medical curriculum, identifies itself distinctly different from the other preclinical or clinical subjects in knowledge base as well as learning/teaching instructions. It exists in series with other pre-clinical courses (e.g., anatomy, biochemistry and physiology), and in parallel with other paraclinical courses such as pathology, microbiology and community medicine. Such arrangement makes learning of pharmacology rather difficult and deficient with regard to its therapeutic relevance and clinical application. In recent years, medical curricula based on clinical cases have emerged as a platform in which pharmacology is one integrated component in a holistic approach to medical education. In this problem-based learning (PBL) model, students learn, with teachers' facilitation, in a student centered environment, based on self-directed, clinically relevant and case oriented approach, usually in a small-group tutorial format. In PBL, pharmacology is learned in concert with other subject issues relevant to the case-problem in question, such as anatomy, physiology, pathology, microbiology, population health, behavior science, etc. Students learn via problem-evoked curiosity and motivation, in an environment which encourages free inquiries and intensive discussions in a cooperative rather than competitive atmosphere. Teachers facilitate students' learning objectives rather than deliver pre-packed knowledge and dictate what they think students should learn. A change towards PBL curriculum appears to be beneficial in better preparing the medical students as life-long learners capable of coping with changes in knowledge and skills associated with the progressive and dynamic social/economic transformation in the Asia-Pacific regions. Evidence is presented that this is indeed happening. PMID- 15339397 TI - Teaching pharmacology to medical students in an integrated problem-based learning curriculum: an Australian perspective. AB - The world-wide move away from the didactic teaching of single disciples to integrated Problem-based Learning (PBL) curricula in medical education has posed challenges for the basic sciences. In this paper we identify two major challenges. The first challenge is the need to describe a core disciplinary curriculum that can be articulated and mapped onto the new structure. We illustrate how the British Pharmacological Society (BPS) Guidelines are used to evaluate the curriculum coverage in the medical course at The University of Melbourne. The second challenge is to ensure that foundational concepts are given adequate emphasis within the new structure, and in particular, that students have the opportunity to pursue these concepts in their self-directed learning. We illustrate one approach to teaching important pharmacological concepts in an integrated curriculum with a case study from the first year curriculum at The University of Melbourne. Finally, we propose the features of an integrated curriculum that facilitates the learning of basic pharmacology in a situation where PBL and integration sets the curriculum framework. PMID- 15339398 TI - A pharmacologist's journey in medical education: a personal history. PMID- 15339399 TI - Teaching of pharmacology in Universiti Malaya and the other medical schools in Malaysia -- a historical perspective. AB - Traditional pharmacology teaching has focused more on drug instead of therapeutics, such that although pharmacological knowledge is acquired, practical skills in prescribing remain weak. In Malaysia many new medical schools (both public and private) have been set up in the last 12 years due to a change in government policy, resulting in a wide spectrum of medical curricula. Universiti Malaya (UM) being the oldest medical school in Malaysia was deep set in its traditional way of teaching-learning, since its inception in 1962, until a visit from the General Medical Council of the United Kingdom in 1984 triggered off a change of tide. Since then the medical curriculum in UM has undergone two major revisions. The first revised curriculum (1988) aimed to inject more clinical relevance into basic science teaching, through introducing clinical lectures and skills in the paraclinical year. Professional behaviour was also addressed. The second revised curriculum (1998) sought to improve further the integration of knowledge as well as to produce a holistic doctor, viewing the patient as a person instead of a clinical entity. The teaching-learning of pharmacology has gradually moved from factual regurgitation to more clinical reasoning, from lab based to more patient-oriented approach. As more new medical schools are being set up in Malaysia, exchange of experience in this area of learning will hopefully help us find a happy medium between "the old is best" and "the new is better" type approach so that a pedagogically sound and yet logistically practical curriculum can be found in our local setting, to help produce doctors with good prescribing practice. PMID- 15339400 TI - Transition from traditional to innovative teaching in and beyond pharmacology at Ziauddin Medical University. AB - Innovative teaching methodologies in different parts of the world are being practiced since last 3 decades. The aim of this review is to report the transition from traditional to innovative self learning process in Ziauddin Medical University, a new medical institute of Pakistan. Various problems encountered have been duly looked after by inducting faculty training programs and regular review sessions in which monitoring of the transition process was duly observed as well as further advancements were also planned. The University being a pioneer has attracted other institutions which have also planned to induct PBL in their curriculum. Vertical and horizontal integration has also inducted better understanding of subjects and fruitful advantages had been accomplished in subjects as pharmacology and pathology. In conclusion, the initial decision of the university to adapt innovative teaching methodologies along with following an integrated curriculum based on PBL and Community Oriented Medical Education system has now made ZMU a Novel institute amongst other medical institutes in the country. PMID- 15339401 TI - Pharmacology teaching and its reform in China. AB - The general situation of pharmacology teaching in China was introduced and the educational reform in China in recent decade is summarized. The aim of the article is to provide those who are interested in teaching of pharmacology to be acquainted with the teaching of pharmacology, including the teaching of both principles and practice, in China. PMID- 15339402 TI - Problem-based learning in pharmacology: a survey of department heads in Taiwan, China. AB - Problem-based learning (PBL) requires active student participation and the use of clinical cases as a trigger to learn within a given area. It has gained much attention as a pedagogic alternative in the course of reform in medical education due to information overload. From discipline-based consideration, it is interesting to understand the views of department heads of pharmacology about implementing PBL for their medical students. According to a general survey from the heads of the department of pharmacology across medical schools in Taiwan, we found that although serious reservation about the approach remains, many departments indeed look forward to including PBL component in their pharmacology curriculum. PMID- 15339403 TI - [Discussion about treatment of severe acute respiratory syndrome based on syndrome differentiation]. AB - Severe acute respiratory syndrome is an infectious disease caused by a new type of coronavirus. It belongs to the seasonal febrile diseases in traditional Chinese medicine. The prevention and treatment of severe acute respiratory syndrome (SARS) can be under the guidance of the doctrines for treating febrile diseases of traditional Chinese medicine, treatment based on syndrome differentiation, such as syndrome differentiation of triple energizer, syndrome differentiation according to defensive phase, qi phase, nutrient phase and blood phase. During April and May of 2003, 8 cases of SARS were diagnosed in Shanghai, and 6 patients accepted complementary therapy of traditional Chinese medicine, without death case. The only one patient who didn't take glucocorticoid therapy was complementarily treated with traditional Chinese herbs through the whole treating procedure. Upon the successful treatment of the eight cases of SARS in Shanghai, it is demonstrated that the triple-energizer syndrome differentiation and defensive-qi-nutrient-blood syndrome differentiation in traditional Chinese medicine are of high value in treating SARS patients. PMID- 15339404 TI - [Treatment of cough and asthma with blood-letting puncturing and cupping: a report of 3 cases]. PMID- 15339405 TI - [Strategy and approaches of pathological and pathophysiological research in integrated traditional Chinese and western medicine]. AB - Pathology and pathophysiology are sciences studying the laws and mechanisms of the occurrence and development of diseases, linking up the preclinical and clinical medicine. Owing to the different perspectives and ways of thinking, the western medicine and the traditional Chinese medicine developed respectively their independent theoretical, diagnostic and therapeutic systems. Integrative medicine, combining the theories and treatments of both western medicine and traditional Chinese medicine, has become the developing trend of medicine along with the social development. For this reason, pathological and pathophysiological research in integrated traditional Chinese and western medicine is highly significant for revealing the internal relations between the clinical manifestation and the pathological changes, for expounding the causes, conditions, mechanisms and laws of the occurrence and development of diseases. In doing related research, we should combine the disease and the syndrome, combine the macro-level and the micro-level, combine the part and the whole. We should manage to systematize the clinical research, to establish animal models of the syndromes, and to integrate the animal models of syndromes with the clinical characteristics of diseases. We should apply the theories of traditional Chinese medicine to the pathological and pathophysiological research of modern medicine. PMID- 15339406 TI - [Pondering acupuncture treatment on anxiety disorders]. AB - Review of papers about acupuncture treatment on anxiety disorders since 1996 showed that many papers were mainly concerned with the treatment of anxiety disorders and obsession with electroacupuncture therapy. Baihui, Yintang, Shenmen, Dazhui, Taiyang, etc, were the most commonly involved acupoints. This paper argues that more emphasis should be transferred from "heart" to "brain", and correspondingly, should be put on acupoints at the head, the DU meridian and acupoints nearby in terms of the position of treatment, on scalp acupuncture and acupoint injection in terms of the methods of treatment, on the combination of diagnosis by western medicine and syndrome differentiation of traditional Chinese medicine in terms of treating strategy. As to the research method, evidence-based medicine, and multicenter, large sample randomized controlled trial study on anxiety disorders should be made. PMID- 15339407 TI - [Typing of bronchiectasis according to syndrome differentiation]. AB - OBJECTIVE: To study the general law of typing of bronchiectasis according to syndrome differentiation. METHODS: We collected the symptoms, conditions of tongue and pulse in patients of bronchiectasis, using frequencies procedure, discriminant analysis and K-means cluster analysis in SPSS statistical software as research medium. RESULTS: Five hundred and sixty three patients with bronchiectasis were studied. It suggested that accumulation of phlegm-heat in the lungs (45.65%), liver fire attacking the lungs (24.51%), asthenia of pulmonosplenic qi (22.38%), asthenia of both qi and yin (7.46%) were the main types. CONCLUSION: Clinical epidemiology provided scientific basis for further studying of the typing of bronchiectasis according to syndrome differentiation. Building up differentiation of syndromes through differentiation and analysis of main symptoms can be used in clinical diagnosis. PMID- 15339408 TI - [Effect of Baisuifang Granule on cognitive malfunction after cerebral infarction]. AB - OBJECTIVE: To explore the mechanism of Baisuifang Granule in treating cognitive malfunction after cerebral infarction. METHODS: One hundred and sixty patients with cerebral infarction were divided randomly into two groups. Eighty patients were treated with Baisuifang Granule and 80 with nimodipine for two months. Clinical observation and laboratory examinations were performed for Mini-Mental State Examination (MMSE), clinical symptoms, Chinese Stroke Scale (CSS), hemorrheological indexes and fibrinogen before and after the treatment. RESULTS: Baisuifang Granule could improve MMSE, reduce the scores of clinical symptoms and CSS, and meliorate the blood rheology. The total effective rate for clinical symptoms in the Baisuifang treated group accounted to 76.25%, with statistical difference comparing to 58.75% of nimodipine treated group (P<0.05). There was significant difference in symptom integral, CSS and whole blood viscosity at the high shear rate, respectively (P<0.01). CONCLUSION: Baisuifang Granule is an effective Chinese medicine for treating cognitive malfunction after cerebral infarction. PMID- 15339409 TI - [Effects of Fuzheng Yiliu Granule on expression of CD44v6 and nm23-H1 in esophageal carcinoma treated with radiotherapy]. AB - OBJECTIVE: To evaluate the effects of Fuzheng Yiliu Granule on the expression of CD44v6 and nm23-H1 in esophageal carcinoma treated with radiotherapy. METHODS: Sixty-three cases of esophageal carcinoma were randomly divided into two groups: Fuzheng Yiliu Granule plus radiotherapy treated group (n=30) and radiotherapy treated group (n=33). The carcinoma specimens were obtained through endoscopic biopsy before and after twenty-one days of treatment. The expression of CD44v6 and nm23-H1 was determined in the sixty-three specimens of esophageal carcinoma by immunohistochemical SABC methods. RESULTS: After twenty-one days of treatment, the positive rates of CD44v6 in the Fuzheng Yiliu Granule plus radiotherapy treated group and the radiotherapy treated group were 40.0% and 69.7%, respectively, and there was significant difference between the two groups (P<0.05). The positive rates of nm23-H1 in the Fuzheng Yiliu Granule plus radiotherapy treated group and the radiotherapy treated group were 20.0% and 21.2%, respectively, and the difference was not significant between the two groups (P>0.05). CONCLUSION: Fuzheng Yiliu Granule can lower the expression of CD44v6 in esophageal carcinoma, and may prevent infiltration and lymph node metastasis of esophageal carcinoma. PMID- 15339410 TI - [Tongue conditions and constitution]. PMID- 15339411 TI - [Clinical observation on treatment of nonalcoholic fatty liver disease complicating hyperuricemia by Zhifang I Decoction]. AB - OBJECTIVE: To observe the therapeutic effect of Chinese herbal recipe Zhifang I Decoction on nonalcoholic fatty liver disease (NAFLD) complicating hyperuricemia (HUA). METHODS: Forty-six patients suffering from NAFLD complicating HUA were randomly divided into treatment group (25 cases) with Zhifang I and control group (21 cases) with Xuezhikang Capsule. One course of treatment was 8 weeks. The data were processed by SPSS 11.0 statistical package after 2 courses of treatment. RESULTS: The total effective rate of the treatment group was 80.00%, which surpassed the control group (71.43%) (P<0.01); Zhifang I could improve the image of B-mode ultrasonography and was better than Xuezhikang in ameliorating the clinical symptoms (P<0.05); Zhifang I could significantly decrease the serum uric acid (UA) (P<0.01), while Xuezhikang had no obvious therapeutic effect on it (P>0.05); Zhifang I was as good as Xuezhikang in recovering alanine aminotransferase (ALT), aspartate aminotransferase (AST),gamma glutamyltransferase (gamma-GT), total cholesterol (TC) and triglyceride (TG) (P>0.05). CONCLUSION: Zhifang I has good effect in treating NAFLD complicating HUA. PMID- 15339412 TI - [Observation by color Doppler ultrasound on endothelium-dependent dilation of congestive heart failure treated with Kanli Decoction]. AB - OBJECTIVE: To observe the changes of endothelium-dependent dilation (EDD) of patients with congestive heart failure (CHF) before and after Kanli Decoction treatment. METHODS: Thirty-five CHF patients were treated with Kanli Decoction. We used color Doppler ultrasound to monitor the flow-mediated dilation (FMD), pulsatile index (PI), resistance index (RI), baseline blood flow (BF), ejection fraction (EF) before and after the treatment. RESULTS: FMD after treatment (9.26+/-3.19) were obviously higher than that before the treatment (7.06+/-2.58) (P<0.05), but BF, PI and RI were not improved. CONCLUSION: Applying color Doppler ultrasound into observation on EDD of patients suffering from CHF and treated by the Kanli Decoction shows that the CHF could be treated by traditional Chinese medicine. PMID- 15339413 TI - [Treatment of malignant tumor with Shenqi Fuzheng injection combined with chemotherapy: a report of 48 cases]. PMID- 15339414 TI - [Effects of Naoluo Xintong Recipe on expression of HSP70 and bFGF in focal ischemia-reperfusion rats]. AB - OBJECTIVE: To investigate the possible beneficial effects of administration of Naoluo Xintong Recipe (NLXT) on the expression of HSP70 and bFGF in rats subjected to ischemia-reperfusion with qi deficiency and blood stasis syndrome. METHODS: We blindly randomized 40 male Wistar rats into 5 groups which were categorized as follows: (1); sham-operated group; (2) rats subjected to transient middle cerebral artery (MCA) occlusion with qi deficiency and blood stasis syndrome (control group); (3) NLXT-treated group; (4) NLXT-pretreated group; (5) Nao'an Capsule-treated group. Focal brain ischemia was induced by the intraluminal suture MCA occlusion method after qi deficiency and blood stasis syndrome had been made. The rats' brains were harvested at the 24h of reperfusion after 2h of MCA occlusion. The expression of HSP70 and bFGF was measured by by immunohistochemical staining. RESULTS: Strong HSP70 immunoreactivity in neurons was observed in NLXT-treated and NLXT-pretreated groups as compared to control group, and increased bFGF immunoreactivity in neurons was observed in the ischemic hemisphere in NLXT-pretreated group in comparison with the ischemic hemisphere in the control group (P<0.01). NLXT has stronger effects on HSP70 and bFGF than Nao'an Capsule. CONCLUSION: The neuroprotective effects of Naoluo Xintong Recipe may be related to its up-regulation of HSP70 and bFGF in the rat brain after focal cerebral ischemia. PMID- 15339415 TI - [Expression of TIMP-1 and TGF-beta1 mRNA in hepatic fibrosis rats and the therapeutic effects of Bushen Rougan Recipe]. AB - OBJECTIVE: To study the expression of TIMP-1 and TGF-beta1 mRNA in hepatic fibrosis rats and the therapeutic effects of Bushen Rougan Recipe (BSRGR). METHODS: Hepatic fibrosis was induced in rats by dimethylnitrosamine (DMN). The rats' hepatic tissue was studied by HE staining and Sirius red staining. The rats were divided into normal control group, fibrosis model group and BSRGR treated group (10 g.kg(-1).d(-1), i.g. for 4 weeks). At last, TIMP-1 and TGF-beta1 mRNA were detected by RT-PCR. RESULTS: Pathological study showed that hepatic fibrosis was successfully induced by DMN in rats. The expression quantity of TIMP-1 and TGF-beta1 mRNA were the most in the fibrosis model group, the second in the BSRGR treated group, and the least in the normal control group. CONCLUSION: The expression of TIMP-1 and TGF-beta1 mRNA was increased in the hepatic fibrosis rats, and this is one possible mechanism of hepatic fibrosis. BSRGR can inhibit the advancement of hepatic fibrosis. PMID- 15339416 TI - [Clinical observation on Biyan Qingdu Granule and ambroxol hydrochloride in treating secretory otitis media]. PMID- 15339417 TI - [Effect of soybean isoflavones on heart function of rats with adriamycin-induced heart failure]. AB - OBJECTIVE: To observe the protective effect of soybean isoflavones (SI) on the heart function of the rats with adriamycin induced heart failure. METHODS: Thirty adult male SD rats were divided into 5 groups:normal control (NC) group, adriamycin (ADR) group, L-SI group, M-SI group and H-SI group. SI of 30, 60, 120 mg.kg(-1).d(-1) was orally administered through a stomach tube once a day for 6 days in L-SI group, M-SI group and H-SI group, respectively. The other two groups were given the same amount of normal saline the same way. Then ADR of 10 mg/kg was given intraperitoneally once to copy the model of heart-failure. The MedLab U/4c biological signal collecting system was used to record and analyze the LVSP of the rats. The pathological changes of the cardiomyocytes were observed. RESULTS: As compared with NC group, the LVSP,+/-dp/dt max, Vpm of the ADR group were significantly lower (P<0.05 or P<0.01), but those of the H-SI group were markedly higher than those of the ADR group (P<0.01). Electron microscopic morphometry of the heart samples of the rats in ADR group revealed typical alterations, consisting an increase of collagen content, vacuolation, diminishing of the cardiomyocyte diameter, alteration of myofilaments and Z-lines of myofibers, and myofibrillar degeneration. SI of 120 mg.kg(-1).d(-1) treatment could prevent the loss of myofibrillae and the reduction of myocyte diameter, and the degeneration of myofilaments and Z-lines were reversed by SI. CONCLUSION: SI of 120 mg.kg(-1).d(-1) treatment can relieve the toxic effect of ADR on myocardium, and also obviously improve the cardiac contractility of heart-failure rats. PMID- 15339418 TI - [Effect of Chinese herbal medicine 1023 Recipe in blocking cancer transformation of experimental precancerous lesion and its mechanism]. AB - OBJECTIVE: To study the effect of Chinese herbal medicine 1023 Recipe in blocking cancer transformation of experimental oral precancerous lesion and its mechanism. METHODS: We treated the experimental oral precancerous lesion in hamster's cheek pouch using 1023 Recipe (consisting of Radix Astragali, Gynostemma Pentaphyllum, Rhizoma Chuanxiong and selenium-rich green tea) for 6 weeks, and observed its effect in blocking cancer transformation, detected 2 kinds of agglutinin receptors (receptors of wheat germ agglutinin and Ricinus communis agglutinin) in the mucosa of the hamster's cheek pouch. RESULTS: The rate of cancer transformation in 1023 Recipe treated group was lower than that in the control group without treatment (P<0.05). Agglutinin receptors in the two groups were different significantly. CONCLUSION: 1023 Recipe is effective in treating hyperplasia, and can prevent its cancer transformation. The mechanism may be that 1023 Recipe can induce precancerous lesions to differentiate into normal tissues. PMID- 15339419 TI - [Experimental study of Qiyeling Decoction inducing apoptosis of transplanted human lung adenocarcinoma cells A549 in nude mice]. AB - OBJECTIVE: To study the function of Qiyeling Decoction in inducing apoptosis of transplanted human lung adenocarcinoma cells A549 in nude mice. METHODS: Nude mice with transplanted A549 tumor were randomly divided into the untreated control group (group A), chemotherapy treated group (group B), chemotherapy plus Qiyeling Decoction treated group (group C), Qiyeling Decoction treated group (group D) and managed correspondingly. The tumor volume was measured and calculated into tumor weight. The apoptosis of tumor cells were examined using in situ cell apoptosis detection kit. RESULTS: The tumor weight was lower obviously in groups B, C and D than that in group A (P<0.05). The apoptosis of tumor cells was lower obviously in groups B, and C than that in group D (P<0.05). Cells in group A appeared perfect differentiation during the early stage and apoptosis later. CONCLUSION: Qiyeling Decoction can induce A549 cell apoptosis in nude mice. PMID- 15339420 TI - [An experimental study on distribution of musk into the brain through blood brain barrier]. AB - OBJECTIVE: To study the possible pathway of the effect of musk on brain disorder, distributing into the brain through blood brain barrier. METHODS: We used the musk ketone (muscone), a main composition of musk, to inject through the tail vein of the rats into the blood and took the brain and other organs at different times to make samples. Then gas chromatography was used to measure the distribution of muscone in the brain and other organs. RESULTS: Muscone could pass through the normal rat's blood brain barrier into the brain and soon reached the highest peak and remained in higher concentration, and more slowly metabolized as compared with other organs. CONCLUSION: Musk distributing into the brain through blood brain barrier provides the basis for its effect in treating brain disorders. Chromatography is an effective method to study the active composition of Chinese herbal medicine distributing through the blood brain barrier into the brain. PMID- 15339421 TI - [Effects of three different drying methods on extraction and separation of ginsenosides from fresh ginseng]. AB - OBJECTIVE: There exists a close relationship between drying of a fresh herb and its preservation and extraction of efficient components. In order to investigate the influences of different drying methods on extraction and separation of ginsenosides, three drying processes, such as drying in the sun, drying in oven and microwave drying, were used to dry fresh ginsengs. METHODS: The ginsenosides of the dry ginsengs were extracted by poaching and microwave heating, and were separated by foam separation. The concentrations of ginsenosides were measured. RESULTS: Microwave drying saved both time and labor, and was favorable for release of ginsenosides. The ginsenosides could be extracted from the dry ginsengs in a shorter time by microwave heating than poaching. The ginsenosides Rb1, Rb2, Rd could be observably concentrated by foam separation. CONCLUSION: Microwave drying and microwave assisted extraction are efficient and economic methods with a high recovery yield of ginsenosides. PMID- 15339422 TI - [A brief talk on pulmonary obstructive syndrome and pulmonary-fibrosis]. PMID- 15339423 TI - [Professor Tang Han-Jun's thinking in syndrome differentiation and experience in treating breast cancer]. PMID- 15339424 TI - [Status of study about Chinese medicine in protecting cerebral ischemia injury]. PMID- 15339425 TI - [Progress in experimental research on apoptosis of hepatic cancer cells induced by Chinese herbs]. PMID- 15339426 TI - [Treatment of diabetes through liver syndrome differentiation]. PMID- 15339427 TI - [Accidents in acupuncture treatment: history and current state]. AB - Acupuncture is widely used as an alternative therapy with few side effects because of its simple manipulation and low cost. However, accidents may occur if the practitioner uses it improperly. Early in the Qin dynasty, the Canon of Medicine mentioned that improper use of acupuncture could cause injury to the body and even death, and it systematically described the occurrence and prevention of acupuncture accidents. Physicians in the successive dynasties frequently reported it, and realized that the manipulation skills were significantly important in preventing the occurrence of acupuncture accidents. Since the 1950s, acupuncture accidents had been effectively prevented because of the improving of acupuncture instruments, enhancing of the quality of the practitioners, popularizing of sterilization and disseminating of anatomic knowledge. Nevertheless, with the renovating of acupuncture techniques, new accidents may occur constantly. The prevention of acupuncture accidents still should be an arduous task for acupuncturists. Nowadays, acupuncture therapy is being used in more than 140 countries, and acupuncture accidents due to improper application are increasing. Prevention of acupuncture accidents has become a global issue deserving of great attention. PMID- 15339428 TI - [Acquirement for researchers of English translation of traditional Chinese medicine]. PMID- 15339429 TI - [Influence of translator's knowledge on the quality of English translation of traditional Chinese medicine]. PMID- 15339430 TI - [Chinese culture and English translation of traditional Chinese medicine]. PMID- 15339431 TI - [Some thoughts about national science and technology award of 2003]. PMID- 15339432 TI - [Treatment of senile osteoporosis with Bushen Zhuanggu Decoction: a report of 32 cases]. PMID- 15339433 TI - [Study on the mechanism of traditional Chinese medicines in promoting gastrointestinal peristalsis]. AB - Traditional Chinese medicine has accumulated rich experience in treating dysfunction of gastrointestinal peristalsis. In recent years, a large number of studies have been made on the mechanism and effects of traditional Chinese medicines on the gastrointestinal peristalsis, and the concept of "gastrointestinal promoting Chinese medicine" has been advocated. These traditional Chinese medicines can be divided into three types: promoting the gastrointestinal peristalsis, inhibiting the gastrointestinal peristalsis, and bi directional modulating. The in vivo and/or in vitro experiments showed that some of the traditional Chinese medicines for activating blood or regulating qi could promote the stomach peristalsis, and the traditional Chinese medicines for moistening intestines to relieve constipation or invigorating spleen to promote digestion could accelerate the intestinal peristalsis. The mechanism lies in the neuroregulation and gut-peptide regulation. Further research on multi-regulation and of multi-target should be done, for the mechanism of the traditional Chinese medicines in regulating the gastrointestinal peristalsis is far more complicated. PMID- 15339434 TI - [New strategy for treatment of respiratory infection and the predominance of traditional Chinese medicine]. AB - Respiratory infection is a common illness. Antibiotic therapy is an essential treatment in clinical practices, but it is challenged by drug-resistant pathogenic microbes. Nowadays, there are many great improvements in some novel non-antimicrobial therapies for preventing and treating respiratory infection, such as enhancing the secretion of endogenous antibiotic peptides to improve innate immune defense of mucous membrane, attaching importance to research and application of vaccine over again, reducing the adhesion and attachment of microbes to block their invasion, and treating systemic inflammatory response syndrome resulting from severe infection. In Chinese herbal pharmacological studies, we found some herbs have antimicrobial function, and furthermore, these herbs have potential effects of immunoregulation. Chinese herbs are worthy of tapping potentialities of research and exploiture. PMID- 15339435 TI - [Relationship between glucocorticoid receptor and deficiency syndrome and the regulation of traditional Chinese medicine]. AB - The glucocorticoid receptor (GR) is a ligand-dependent transcription factor belonging to the nuclear hormone receptor superfamily. Due to its almost ubiquitous expression, GR plays an important role in many physiological and pathological processes. These include regulation of homeostasis, adaptation to stress, and modulation of central nervous system. In addition, GR is a major modulator of the immune system due to its proficient anti-inflammatory and immunosuppressive activity; and its function is important for proper regulation of many physiological processes. The exploration on the regulation of traditional Chinese medicine on GR has established a modern physiological and pathological foundation for research of traditional Chinese medicine. PMID- 15339436 TI - [Characteristics of sublingual venae in primary liver cancer patients in different clinical stages]. AB - OBJECTIVE: To explore the characteristics of the sublingual venae in patients with primary liver cancer (PLC). METHODS: The shape, color and the red, green and blue values of the sublingual venae were analyzed quantitatively for PLC patients in different clinical stages by analysis system for comprehensive information of tongue diagnosis. RESULTS: With the aggravating of the disease, the patients' sublingual venae became wide and tortuous, and their color became blue and purple. The abnormality of the sublingual venae was more serious in clinical stage III than in stages I and II (P < 0.05). CONCLUSION: The shape, color and abnormality degree of the sublingual venae in patients with PLC in different clinical stages are distinct. PMID- 15339437 TI - [Portal hemodynamics in patients with different syndromes of cirrhosis]. AB - OBJECTIVE: To explore the relationship between the portal hemodynamics or the indices of liver fibrosis and the liver function score in patients with different traditional Chinese medical syndromes of liver cirrhosis. METHODS: One hundred and forty-seven cases of liver cirrhosis with different syndromes were included in the study. Diameters and blood flow velocities of the portal vein and splenic vein were tested by color Doppler sonarography. The indices of liver function (TBIL, ALT, AST, Alb, Glb, PTA) and the indices of hepatic fibrosis (HA, PC-III, LN, VI-C) were tested. RESULTS: The diameters of portal vein in liver cirrhosis patients with different syndromes (internal accumulation of of damp-heat, stagnation of liver-qi, superabundance of dampness due to spleen-asthenia, asthenia of liver and kidney yin, asthenia of spleen and kidney yang, blood stasis) were not significantly different. The blood flow velocities and flow volumes of portal vein in patients with the blood stasis syndrome and the internal accumulation of damp-heat syndrome were significantly different (P < 0.05). The diameter of splenic vein in patients with the internal accumulation of damp-heat syndrome was significantly different to that of the asthenia of liver and kidney yin syndrome, or the blood stasis syndrome, or the asthenia of spleen and kidney yang syndrome, respectively (P < 0.05 or P < 0.01). The blood flow velocity of splenic vein in patients with the blood stasis syndrome or the asthenia of spleen and kidney yang syndrome was significantly different to that of the internal accumulation of damp-heat syndrome, or the stagnation of liver-qi syndrome, or the asthenia of liver and kidney yin syndrome, respectively (P < 0.01). The blood flow volume of splenic vein in patients with the internal accumulation of damp-heat syndrome was significantly different to that of the asthenia of liver and kidney yin syndrome, or the asthenia of spleen and kidney yang syndrome, or the blood stasis syndrome, respectively (P < 0.01). The liver cirrhosis indices HA and PC-III in patients with the asthenia of liver and kidney yin syndrome were significantly different to those of the internal accumulation of damp-heat syndrome (P < 0.05). In Child-Pugh classification, the incidence of Child-Pugh C was the highest in the asthenia of spleen and kidney yang syndrome, while the incidence of Child-Pugh A was the highest in the stagnation of liver qi syndrome. CONCLUSION: The changes of portal vein dynamics in liver cirrhosis patients with the asthenia of liver and kidney yin syndrome, or the asthenia of spleen and kidney yang syndrome, or the blood stasis syndrome are significantly different to those of the internal accumulation of damp-heat syndrome and the stagnation of liver-qi syndrome. PMID- 15339438 TI - [Clinical observation on treatment of cervical spondylosis of vertebral artery type with Jingkang Decoction]. PMID- 15339439 TI - [Influence of Jieyu Huoxue Decoction on rehabilitation of patients with depression after cerebral infarction]. AB - OBJECTIVE: To study the influence of Jieyu Huoxue Decoction on rehabilitation of patients with depression after cerebral infarction. METHODS: Fifty four patients suffered from the depression after acute cerebral infarction were randomly divided into three groups: depression control group, fluoxetine treated group and Jieyu Huoxue Decoction treated group. Eighteen patients of cerebral infarction without depression were enrolled in the no depression control group randomly. The Zung self-rating depression scale (SDS), the modified Edinburgh-Scandinavia stroke scale (MESSS) and the activities of daily living (ADL) were evaluated before treatment, 30 and 60 days after treatment. RESULTS: After 30 and 60 days of treatment, SDS evaluation of the Jieyu Huoxue Decoction treated group showed significant difference as compared with the condition before treatment and that of the depression control group (P < 0.05 or P < 0.01), MESSS evaluation and ADL evaluation also showed significant difference as compared with the condition before treatment and that of the depression control group (P < 0.05 or P < 0.01); there was no obvious difference between the fluoxetine treated group and the Jieyu Huoxue Decoction treated group, but the Jieyu Huoxue Decoction treated group showed fewer symptoms and less side effect. CONCLUSION: Jieyu Huoxue Decoction can not only relieve depression after cerebral infarction, but also improve neurological functions. PMID- 15339440 TI - [Evaluation of clinical reports in treating periarthritis of shoulder by manipulation with the theory of evidence based medicine]. AB - OBJECTIVE: To explore the effect of treating periarthritis of shoulder with manipulation through evaluating clinical reports with the theory of evidence based medicine (EMB). METHODS: By retrieval of some main medical literature databases in the past 5 years, 242 articles on treating periarthritis of shoulder with manipulation were available. Selected by admission standard, 10 of them were evaluated with EBM theory. RESULTS: Both single-application and union-application of manipulation were effective to periarthritis of shoulder, but the evaluated articles were all low-quality articles. CONCLUSION: Because of low quality clinic test and larvaceous publication bias, the extant evidence is not enough to recommend replacing other therapeutic methods by manipulation. The low quality of evaluated articles suggests that there exist some methodology problems in the clinic test on treating periarthritis of shoulder with manipulation. PMID- 15339441 TI - [Intensive effect of traditional Chinese medicines activating blood to resolve stasis on medicines dredging intestines--influence on peristalsis of small intestine in guinea pigs]. AB - OBJECTIVE: To observe the changes of peristalsis of small intestine in guinea pigs after administration of traditional Chinese medicines activating blood to resolve stasis (Compound Danshen Decoction, CDSD) or/and medicines dredging intestines (Dachengqi Decoction, DCQD), and to explore the synergetic or intensive effect of CDSD on DCQD. METHODS: By means of BL-420 Biological Experimental System, peristalsis of small intestine was recorded and analyzed following administration of DCQD, CDSD or Huoxue Chengqi Decoction (HXCQD, compound of CDSD and DCQD) respectively in different experimental periods. RESULTS: The amplitude and frequency of intestinal peristaltic wave obviously increased following administration of the three decoctions, but HXCQD appeared to be most dominantly. CONCLUSION: The effect of DCQD can be further enhanced by combining use of CDSD, suggesting that the traditional Chinese medicines activating blood to resolve stasis have an intensive effect on medicines dredging intestines. PMID- 15339442 TI - [Effects of salidroside on carbohydrate metabolism and differentiation of 3T3-L1 adipocytes]. AB - OBJECTIVE: To observe the effects of salidroside on carbohydrate metabolism and differentiation of 3T3-L1 adipocytes. METHODS: 3H-glucose uptaking rate in adipocytes treated by salidroside was detected. Adipocytes treated by salidroside in differentiation were stained by oil red O and analyzed by spectrophotography quantitatively. The expression of PPAR-gamma and C/EBP-alpha mRNA relating to differentiation of adipocytes was detected by reverse transcription PCR. RESULTS: 3H-glucose uptaking rate in the salidroside group was 110.4%, higher than that in the blank control group (P < 0.01). Salidroside suppressed the differentiation of 3T3-L1 pre-adipocytes and down-regulated the expression of PPAR-gamma and C/EBP alpha mRNA (P < 0.01, vs control group). CONCLUSION: Salidroside promotes the 3H glucose uptaking, suppresses the differentiation and down-regulates the expression of PPAR-gamma and C/EBP-alpha mRNA in 3T3-L1 adipocytes. PMID- 15339443 TI - [Effect of recipes replenishing qi and activating blood on cell proliferation and apoptosis in the liver of aging rats]. AB - OBJECTIVE: To observe the effect of recipes replenishing qi and activating blood on cell proliferation and apoptosis in the liver of natural aging rats. METHODS: Natural aging rats were under administration of recipes replenishing qi or activating blood for 4 months. The liver of the rats was prepared into cell suspension for determination of cell proliferation and apoptosis with PI-staining and flow cytometer. RESULTS: (1) Compared with those of the young rats, the cells in G(0)-G1 phase in the liver tissue of aging rats were increased (P< 0.01), and apoptosis cells were increased (P< 0.01), while the cells in S and G2-M phases were decreased (P< 0.01). (2) Compared with those of the aging rats, the cells in G(0)-G1 phase in the liver tissue of aging rats administered recipes replenishing qi or activating blood were decreased (P< 0.01), and it was more obvious in activating blood group than in replenishing qi group (P< 0.01); the cells in S and G2-M phases were increased (P< 0.01) and there was no significant difference between the activating blood group and the replenishing qi group (P> 0.05). (3) The apoptosis cells in replenishing qi or activating blood group were decreased significantly (P< 0.01), and the effect of replenishing qi was better than that of activating blood (P< 0.01). CONCLUSION: (1) Cell proliferation is decreased and apoptosis is increased in the liver tissue of natural aging rats. (2) Recipes replenishing qi or activating blood can accelerate cell proliferation in the liver tissue of natural aging rats, and the effect of activating blood was slightly stronger than that of replenishing qi. (3) Recipes replenishing qi or activating blood can inhibit cell apoptosis in the liver tissue of natural aging rats, and the effect of replenishing qi was better than that of activating blood. PMID- 15339444 TI - [Influence of single leaf Asarum himalaicum on renal function of rabbits]. AB - OBJECTIVE: To study the influence of single leaf Asarum himalaicum on the renal function of rabbits. METHODS: Rabbits were divided into three groups. Asarum himalaicum, Asarum heterotropoides and normal saline were intravenously administered to the rabbits of one group respectively. The urine volume per minute, urine pH, urine glucose, protein and red blood cells, BUN, SCr, TXB2, 6 keto-PGF1alpha, TXB2/6-keto-PGF1alpha, endothelin, p-aminohippuric acid clearance rate and phenolsulfonphthalein excretion rate were tested before and after the administration. RESULTS: A certain dosage of single leaf Asarum himalaicum caused acute renal failure in rabbits. The indices tested were significantly different between rabbits administered Asarum himalaicum and normal saline. As compared with the rabbits administered Asarum heterotropoides, the results of indices tested decreased, but without statistical significance, except for SCr. CONCLUSION: The single leaf Asarum himalaicum can cause renal damage to rabbits. Its renal toxicity is lower that that of Asarum heterotropoides. PMID- 15339445 TI - [Effect of Qinggong Zhixue Granule on low immunity mice]. AB - OBJECTIVE: To probe into the pharmacological mechanism of the Chinese medicinal compound Qinggong Zhixue Granule (QGZXG), which has an effect of replenishing qi to activate blood and expelling stasis to stop bleeding, in treating irregular vaginal bleeding after medical abortion. METHODS: Healthy female KM mice and mice with immunodeficiency caused by cytoxan (CTX) were chosen as experimental subjects. The effects of QGZXG on the phagocytization of the mice's monocytes (by using the method of carbon particles expurgation), the delayed type hypersensitivity caused by 2,4-dinitrochlorobenzene (DNCB) and the content of antibody of hemolysin in the mice's serum were observed. RESULTS: (1) QGZXG improved the ability of the liver and spleen of immunodeficient mice to expurgate carbon particles. There was an significant statistical difference between the CTX treated group and the low dose group as well as the high dose group (P < 0.01) and a statistical difference between the CTX treated group and the medium dose group (P < 0.05). (2) QGZXG boosted the cellular immunity of immunodeficient mice. The ear swelling of mice in low, medium and high dose group was more obvious than that in the CTX treated group, and this difference was significant in statistics (P < 0.01). (3) QGZXG raised the content of the antibody of hemolysin in the serum of immunodeficient mice. A statistical difference occurred between the high dose group and the CTX treated group. The low and medium dose also had a tendency of such effect. CONCLUSION: QGZXG can improve both the specific and non-specific immunity of the immunodeficient mice, and therefore accelerate the recovery of the whole body and endometrium after parturition and abortion. In consequence, it leads to shorter duration and less quantity of vaginal bleeding after medical abortion. PMID- 15339446 TI - [Effect of oxymatrine on pathological change in brain tissue of newborn mice infected by cytomegalovirus]. AB - OBJECTIVE: To study the effect of oxymatrine on pathological change in brain tissue of newborn mice infected by cytomegalovirus (CMV). METHODS: CMV of TCID50 was inoculated into the brain of the newborn mice, and the morphological change in the brain tissue infected by CMV was observed with light microscope and transmission electron microscope. RESULTS: In the model control group, the results showed that there were some inflammatory cellular infiltration and focal necrosis in the brain tissue of newborn mice infected by CMV. The ultrastructure change in the brain tissue showed that the nuclear membrane of cerebral neurons sunk, the chromatin deformed and fused into masses, the cytoplasm vacuolated, the endoplasmic reticulum disarranged and the Nissl's body was blurred or disappeared. After being treated with oxymatrine (50 mg/kg, ip) for 15 days, those pathological changes of the brain tissue in the newborn mice could be significantly improved. CONCLUSION: Oxymatrine has an obvious inhibition on CMV in vivo. PMID- 15339447 TI - [Effect of melittin on apoptosis and necrosis of U2 OS cells]. AB - OBJECTIVE: To study the effect of melittin on apoptsis and necrosis of osteosarcoma cell line U2 OS in vitro. METHODS: Osteosarcoma cell line U2 OS was treated with melittin. The growth and proliferation was observed by MTT assay and cell counting, and the necrosis was estimated by Trypan blue staining. The cell apoptsis, Fas and Apo2. 7 expression were detected by cytometer. RESULTS: The data showed that melittin could inhibit the proliferation of U2 OS dose dependently at 16 and 64 mg/L. Cell apoptsis was detected by cytometer, when the cells were treated by 16 mg/L and 32 mg/L of melittin respectively, and the percentages of Fas and Apo2. 7 positive cells were increased. CONCLUSION: Melittin inhibits the proliferation of osterosarcoma cell line through up regulating Fas expression and inducing apoptsis. PMID- 15339448 TI - [Effect of Compound Rhubarb Mixture on contraction of isolated small intestinal smooth muscle of rabbit]. AB - OBJECTIVE: To investigate the effect of Compound Rhubarb Mixture on contraction of isolated small intestinal smooth muscle of rabbit. METHODS: Serum containing Compound Rhubarb Mixture was prepared. The contraction curves of isolated small intestinal smooth muscle before and after administration of the serum were recorded by Medlab biological information collecting system. RESULTS: The serum containing Compound Rhubarb Mixture amplified the contraction amplitude of isolated small intestinal smooth muscle of rabbit. The rate of change of contraction amplitude was elevated significantly after administration, while the frequency of contraction did not change obviously. CONCLUSION: The serum containing Compound Rhubarb Mixture can enhance the contractive function of isolated small intestinal smooth muscle of rabbit. PMID- 15339449 TI - [Inhibition of Longzhi extracts on tumor growth of transplanted H22 and S180 in mice]. AB - OBJECTIVE: To study the inhibitory effect of Longzhi extracts (extracts from a Chinese herbal compound) on the growth of transplanted hepatoma H22 and sarcoma S180 in mice. METHODS: Certain dosage of Longzhi extracts was dissolved into distilled water to obtain the suspension of definite concentration. The suspension of 0.2 ml was planted on the right forefoot of each mouse. The mice were randomly divided into groups 24 hours later and treated for 7 days. One day after the treatment stopped, the mice were sacrificed and the tumor masses were taken out and weighed. RESULTS: The inhibition rates of Longzhi extracts on the tumor of transplanted H22 and S180 were 63% and 41% respectively. CONCLUSION: Longzhi extracts has antitumor action. PMID- 15339450 TI - [A preliminary report on solid-state fermentation of Ganoderma lucidum with Radix Astragali containing medium]. AB - OBJECTIVE: To test the practicability of the solid-state fermentation for medicinal fungi by fermenting Ganoderma lucidum with Radix Astragali containing medium. METHODS: Ganoderma lucidum was fermented in ordinary medium, drug containing medium (containing Radix Astragali) and selenium-rich drug-containing medium respectively. The polysaccharide contents of fermentation products from the three kinds of culture media were tested at different time, and the changes were compared. RESULTS: The polysaccharide contents of fermentation products from the three kinds of culture media were 4.65%, 3.76% and 4.50% respectively and their relative standard deviation were 1.61%, 1.99% and 1.86% respectively. By observing the changes of the contents of polysaccharide, protein and total saponin in fermentation products from the drug-containing medium at different time, it was found that the 28th fermentation day was the time when secondary metabolism was most active, and it should be the fermented terminal point. CONCLUSION: The fermentative combination of Ganoderma lucidum and Radix Astragali is practicable. PMID- 15339451 TI - [Clinical evaluation of electro-acupuncture on sacral points for urinary urge incontinence]. PMID- 15339452 TI - [Clinical observation on manipulation without syndrome differentiation in treating infantile diarrhea]. PMID- 15339454 TI - [Professor Tong Xiao-Lin's experience in syndrome differentiation and treatment for high fever]. PMID- 15339455 TI - [Progress in research on pharmaceutics for intranasal medication]. PMID- 15339456 TI - [Progress in pharmacotherapy of integrated traditional Chinese and western medicine for virus infection in respiratory tract]. PMID- 15339457 TI - [Progress in research on regulation of traditional Chinese medicine on platelet derived growth factor]. PMID- 15339459 TI - [Re-discussion on the translating principles of traditional Chinese medicine]. PMID- 15339458 TI - [Discussion on clinical efficacy of Hypericum perforatum for depression abroad]. AB - The development of molecular pharmacology and neuropharmacology accelerated the studies on molecular mechanism of Hypericum perforatum for depression. The clinical trials indicated that this galenical was superior to the traditional synthetic drugs for antidepression. This preparation had good tolerability and safety. Clinical pharmacokinetics and pharmacodynamics provided further evidence for clinical application of Hypericum. The clinical efficacy of Hypericum for depression was notable and credible. PMID- 15339460 TI - [Exploring into the principles of Chinese-English translation of traditional Chinese medicine]. PMID- 15339461 TI - [Current status of blood lipid level and treatment of hyperlipoidemia in Chinese population]. PMID- 15339462 TI - [Integrated traditional Chinese and western medicine should make new contribution to the reproductive health of women]. AB - The Obstectrics and Gynecology Hospital attached to Fudan University has made great achievements in the clinical and experimental study of integrated traditional Chinese and western medicine in the past 45 years. Study on induction of ovulation with acupuncture revealed the relationship between circulating estrogen and central beta-endorphin, and suggested that the depletion of central beta-endorphin resulted in the release of GnRH and ovulation, and the main indication for acupuncture treatment was pubertal dysfunctional uterine bleeding. Acupuncture was also successfully used to treat anorexia nervosa. The successful treatment of polycystic ovary syndrome (PCOS) with Yu's Tonifying Recipe for tonifying the kidney and resolving phlegm was found to be related to the elevation of serum FSH which resulted in ovulation. Good result was obtained in hyperinsulinemia pattern of PCOS treated with Tiangui Recipe for replenishing the kidney and activating blood. In studies on patients and animal model, Tiangui Recipe was found to decrease peripheral androgen and insulin levels and lead to central regulation of proopiomelanocortin, neuropeptide Y and leptin receptor, as well as regulation of neuro-endocrine-metabolic network. Thus it can promote ovulation and reduce body weight. In the studies, a new way to classify PCOS was suggested. Gengnianchun Recipe for women with postmenopausal syndrome was found to be effective in increasing estrogen receptors (ERs) and ER mRNA in hypothalamus and peripheral organs and regulating neuro-endocrine-immune metabolic network in patients and aging rats. Gongtai was effective for controlling menorrhagia under the mechanism of regulating endometrial prostaglandins. The researches mentioned above indicate that illness is the disorder of some key-links in the life network of the patient which may be affected by factors from the environment. And this idea is different from the ideas in traditional Chinese medicine and biomedicine. It indicates that woman in subclinical health state has already got something incontrollable in her life network, and it also gives a new impact on the treatment and prevention of women's diseases as well as the prevention and treatment of women's reproductive health. PMID- 15339463 TI - [Pondering the research on syndrome of traditional Chinese medicine]. AB - Syndrome research is a challenging field in the research on traditional Chinese medicine and there is no breakthrough at all in this field as yet, although some progresses from various aspects have been achieved in recent years. The definition of syndrome based on the conceptions of traditional Chinese medicine is basically clear, but the qualitative macro-criteria for syndrome are not standardized and the research methods for quantification of such criteria are not unified. These hindered the processing of the modernization of traditional Chinese medicine to a great extent; therefore new research methods for syndrome using techniques of modern sciences should be further investigated. During the research of syndrome, it is suggested that: (1) the research should be turned to complex syndrome from the fundamental simple syndrome gradually, on the basis of certain diseases in western medicine; (2) animal experiment would be of no significance before quite a successful research performed in human body; (3) the results of the research of syndrome should be in agreement with the life science as well as the entire natural science. PMID- 15339464 TI - [Application of DNA microarray technology in immunological research and its inspiration to researches on traditional Chinese medicine]. AB - DNA microarray technology is a high throughout method that can analyze the expression of thousands of genes at the same time to form a complete cellular gene expression profile. It has been applied in immunological researches such as the development, maturation, activation and differentiation of immune cells, the regulation of immune responses, the molecular mechanism of allergy, the relation between phenotype and gene expression, and immunological pharmacology, etc. It has deepened our perception of the immune system. It will as well be helpful in the research of the regulative mechanism of traditional Chinese medicine (TCM) toward immune cells and immune responses, the therapeutic mechanisms of TCM toward allergy, the standardization of differentiation of syndrome and herbal pharmacology, etc. The data analysis of the results of DNA microarray experiments is aimed to compare the difference of gene expression levels of different samples and obtain diagnostic gene expression profiles. But there are controversies about the number of samples and choices of statistical methods. Researchers are striving in setting standards of DNA microarray experiments. PMID- 15339465 TI - [Effect of Xiangdan Injection on mRNA expression of endothelial vaso-active factors of patients with coronary heart disease and blood stasis]. AB - OBJECTIVE: To evaluate the effect of Xiangdan Injection on mRNA expression of the endothelial vaso-active factors of patients with coronary heart disease and blood stasis. METHODS: Fifty-six patients were randomly divided into two groups:twenty eight patients were treated according to the therapeutic guide for coronary heart disease as the control group and 28 were given the same treatment plus Xiangdan Injection as the treated group. The expressions of ET-1 and eNOS mRNA were examined with RT-PCR before experiment and ten days later. RESULTS: The positive rate of eNOS mRNA of the treated group increased, while the positive rate of ET-1 mRNA of the treated group decreased after ten day's treatment, with significant differences as compared with that before the experiment. Xiangdan Injection up regulated the eNOS mRNA expression and suppressed the ET-1 mRNA expression. Changes of expression were not observed in the control group. CONCLUSION: Xiangdan Injection improves the endothelial function of patients with coronary heart disease and blood stasis by regulating the expressions of ET-1 and eNOS mRNA. PMID- 15339466 TI - [Clinical study on Fufang Sishen Decoction in treating arrhythmia after virus myocarditis]. AB - OBJECTIVE: To observe the effect of Fufang Sishen Decoction (FFSSD) on arrhythmia after virus myocarditis. METHODS: One hundred and two cases of arrhythmia after virus myocarditis were randomly divided into two groups. The treatment group was treated with FFSSD, 6 g, b.i.d.; and the control group with propafenone, 150 mg, q 8 h. The therapeutic effects were observed in 4 weeks. RESULTS: The total anti arrhythmia effects of FFSSD and propafenone were 71.9% and 78.9% respectively (P>0.05). FFSSD took effects relatively slowly with mild and lasting effect. CONCLUSION: The curative effect of FFSSD in treating arrhythmia after virus myocarditis is confirmed. FFSSD has no obvious side effects. PMID- 15339467 TI - [Effects of granules for nourishing kidney and strengthening brain on main symptoms of kidney-yin deficiency and blood stasis syndrome and hemorrheological characteristics of brain atrophy]. AB - OBJECTIVE: To explore the influence of granules for nourishing kidney and strengthening brain (GNKSB) on main symptoms of kidney-yin deficiency and blood stasis syndrome and the hemorrheological characteristics of brain atrophy patients. METHODS: Ninety patients of brain atrophy with kidney-yin deficiency and blood stasis syndrome were randomly divided into two groups. Sixty cases in treatment group were treated with GNKSB, and the 30 cases in control group were treated with piracetam for 8 weeks. RESULTS: The effective rate of treatment group was 73.3%, with significant difference as compared with 46.6% of the control group (P<0.01). The scores of symptom-assessment of the two groups were also significantly different (P<0.01). The mini-mental state examination of treatment group was obviously improved, and was significantly different as compared with the control group (P<0.01). The scores of Hasegawa's dementia scale and activities of daily living were increased, but without significant difference as compared with the control group. The platelet aggregation rate was improved, with significant difference as compared with the control group (P<0.05). The whole blood viscosity was also improved obviously, but without significant difference as compared with the control group. CONCLUSION: GNKSB is effective for kidney-yin deficiency and blood stasis syndrome of brain atrophy patients and can improve their mental state and the hemorrheological indexes. PMID- 15339468 TI - [Treatment of non-small-cell lung cancer with chemotherapy and Sansheng Huatan Decoction]. AB - OBJECTIVE: To observe the clinical effect of Chinese medicine Sansheng Huatan Decoction combined with chemotherapy in treating advanced primary non-small-cell lung cancer and to evaluated the effect of Sansheng Huatan Decoction increasing clinical effect and decreasing toxicity in chemotherapy for non-small-cell lung cancer. METHODS: One hundred and sixty patients of advanced primary non-small cell lung cancer proved by pathological examination were randomized into two groups. The treatment group was treated with Sansheng Huatan Decoction and chemotherapy, and the control group was treated only with chemotherapy. The clinic effect, life quality, natural killer (NK) activities, liver and kidney functions, and blood routine test of the 2 groups were evaluated. RESULTS: The clinical effective rates of the treatment and control groups were 56.7% and 48.2% respectively, and there was no statistic significance (P>0.10). The life quality, NK activities and blood routine test of the treatment group were better than those of the control group. CONCLUSION: Sansheng Huatan Decoction combined with chemotherapy is a better treatment for non-small-cell lung cancer as compared with chemotherapy. PMID- 15339469 TI - [Effects of fraction from Phyllanthus urinaria on thrombosis and coagulation system in animals]. AB - OBJECTIVE: To investigate the effects of PUW (a fraction containing 60% corilagin from a Chinese herbal plant Phyllanthus urinaria) on thrombosis and coagulation system. METHODS: Myers's method was used to evaluate the protection of intravenously administered PUW against the male mouse sudden death caused by injection of 75 mg/kg arachidonic acid in the tail vein; Charlton's method was modified to observe antithrombotic effect of PUW in rat electrically stimulated carotid artery thrombosis model; and the model of venous thrombosis was produced to investigate the antithrombotic effect of PUW. Rosette assay was used to observe the effect of PUW on platelet-neutrophil adhesion. The effects of PUW were monitored on euglobulin lysis time (ELT), prothrombin time (PT), kaolin partial thromboplastin time (KPTT), and bleeding time (BT) in rats, according to the methods of Kowalski, HUANG Zheng-Liang, and GU Yue-Fang, et al, respectively. RESULTS: The results showed that PUW administered intravenously significantly decreased the mouse mortality, prolonged the occlusion time of rat carotid arteries, and reduced the wet and dry thrombus weight of the inferior vena cava, respectively. PUW markedly inhibited the binding of activated platelets to neutrophils, obtaining 39.7 mg/L of the medium inhibitory concentration. Intravenously administered PUW significantly shortened ELT, prolonged KPTT while had no influence on PT; PUW increased BT in rat tail tips but the BT caused by PUW was much shorter than that by aspirin or urokinase. CONCLUSION: It is suggested that PUW has the potential of antithrombosis due to its inhibition of platelet-neutrophil adhesion. PUW shows the tendency to bleeding, however, it could not cause serious bleeding side effect as compared with aspirin or urokinase. PMID- 15339470 TI - [Effect of Zhenju Jiangya Tablet on the endothelial function of hyperlipidemic rabbits]. AB - OBJECTIVE: To study the effect of Zhenju Jiangya Tablet (ZJ) on the injured endothelial cells and endothelium-dependent relaxation function of hyperlipidemia rabbits. METHODS: Male New Zealand rabbits were randomized into four groups: control group, hyperlipidemia group, ZJ group and sivastatin group. The endothelium-dependent relaxation function was evaluated by APV using intravascular Doppler, and the morphology of endothelial cells was detected by light microscopy and electron microscopy, and nitric oxide synthase was evaluated. RESULTS: ZJ reduced the lesions of hyperlipidemia vessels, and the APV after Ach injection of each group was (1.14+/-0.26), (1.74+/-0.59), (1.22+/-0.37) and (1.17+/-0.41) respectively. The eNOS of each group was (4.21+/-0.37), (1.43+/ 0.88), (3.95+/-0.67) and (4.08+/-0.46) nmol x min(-1) x g(-1) respectively. CONCLUSION: ZJ can improve the abnormality of endothelial cells and endothelium dependent relaxation function of hyperlipidemia. PMID- 15339471 TI - [Effect of hesperidin and rutin on oxidative modification of high density lipoprotein in vitro]. AB - OBJECTIVE: To study the effect of hesperidin and rutin on oxidative modification of high density lipoprotein (HDL) in vitro. METHODS: HDL was isolated from healthy human plasma by sequential ultracentrifugation, and was oxidized by copper ions. The inhibitory effects of hesperidin and rutin on HDL oxidative modification were valued by the formation of malondialdehyde (MDA). RESULTS: Hesperidin and rutin significantly inhibited copper-induced oxidation of HDL in a dose-dependent manner. CONCLUSION: Both hesperidin and rutin can prevent HDL from copper-induced oxidative modification in vitro. This result suggests that they might have antiatherogenic effect. PMID- 15339472 TI - [Preventive effect of Breviscapus Injection on chronic hypoxic myocardium injury in rats]. AB - OBJECTIVE: To study the preventive effect of Breviscapus Injection on the chronic hypoxic myocardium injury in rats and its mechanism. METHODS: Transmission electron microscope and biochemical analysis were used to assay the therapeutic effect of Breviscapus Injection in rat model induced by 4 week's hypoxia. RESULTS: The contents of malondialdehyde (MDA) and Ca(2+) in the homogenate of myocardial tissue of the hypoxic rats were significantly higher than those of the control rats. The activities of superoxide dismutase (SOD), nitric oxide synthase (NOS) in the homogenate of myocardial tissue and the plasmic NO levels of the hypoxic rats were significantly lower than those of the control rats. Breviscapus Injection reduced the contents of MDA and Ca(2+), and increased the levels of SOD, NOS and NO. Ultrastructural examination revealed that the injuries of the ultrastructure of heart in hypoxic rats were improved by treatment with Breviscapus Injection. CONCLUSION: Breviscapus Injection can effectively prevent and treat the hypoxia-induced myocardial damage. One of its mechanisms may relate to its adjusting NO level, anti-damaging of free radicals and inhibiting calcium overload. PMID- 15339473 TI - [Role of cyclin kinase inhibitor p27 in inhibition of emodin on mesangial cell proliferation induced by tumor necrosis factor-alpha]. AB - OBJECTIVE: To investigate the role of p27 in the inhibition of emodin on the mesangial cell (MC) proliferation induced by tumor necrosis factor-alpha(TNF alpha). METHODS: p27 protein of MC was detected with western blotting analysis. The degree of MC proliferation was estimated through [(3)H] thymidine ([(3)H] TdR) incorporation. Different dosage of emodin (50 mg/L,100 mg/L) was added into MC stimulated by TNF-alpha. RESULTS: TNF-alpha (200 kU/L) decreased p27 level of MC cultured in serum-free DMEM for 24 hours and increased[(3)H] TdR incorporation. Emodin increased p27 level of MC stimulated by TNF-alpha and decreased [(3)H] TdR incorporation. The more the emodin was added, the greater the above-mentioned effect of emodin. CONCLUSION: The increment of p27 level maybe play an important role in the inhibition of emodin on MC proliferation induced by TNF-alpha. PMID- 15339474 TI - [An animal model of alcoholic fatty liver and the prevention and treatment with New Qinggan Decoction]. AB - OBJECTIVE: To establish an animal model of alcoholic fatty liver (AFL) and to observe the prevention and treatment with New Qinggan Decoction (NQGD). METHODS: Female SD rats were exposed to the mixture of white spirit and corn oil continuously for up to 6 weeks via an intragastric feeding protocol. The pathologic alteration of the liver and the changes of serum lipid were observed, and the effect of NQGD for preventing and treating AFL was studied. RESULTS: Wide hepatic steatosis was induced in model group rats. Serum triglyceride (TG) and total cholesterol (TC) of the model group were higher than those of the normal group (P<0.01). Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of the model group were higher than those of the normal group (P<0.05). In NQGD group, hepatic steatosis was milder than that in the model group, and serum TG, TC, AST, ALT descended significantly as compared with the model group (P<0.01). CONCLUSION: AFL model of rats established by feeding the mixture of white spirit and corn oil is similar to the episode of the human AFL, and is easy to intervene with medicine. NQGD can prevent the occurrence of AFL in this model. PMID- 15339475 TI - [Effects of Shendan Jianyi Capsule on aldose reductase activity of kidney in diabetic rats]. AB - OBJECTIVE: To evaluate the effects of Shendan Jianyi Capsule (SDJYC) on aldose reductase (AR) activity of the kidney in diabetic rats and the significance of prevention and treatment of diabetic nephropathy with Chinese natural herbs. METHODS: Diabetic Wistar rats were treated with SDJYC and sorbinil respectively, and the AR activity in kidney, urine protein and the activity of Na(+)-K(+) ATPase were tested. RESULTS: The AR activity in diabetic rats was decreased obviously after being treated with sorbinil or SDJYC. The activity of Na(+)-K(+) ATPase in diabetic rats decreased obviously at the end of the eighth week, and increased after being treated with sorbinil and SDJYC. There was no difference between two treated groups after the treatment. It demonstrated that the inhibition of AR activity could improve the activity of Na(+)-K(+)-ATPase and SDJYC had the same effect. CONCLUSIONS: SDJYC exerts effects by inhibiting the activity of AR. PMID- 15339476 TI - [Hemostatic and analgesic effect of Gonghuan Zhixue Tablet on mice]. AB - OBJECTIVE: To explore the hemostatic and analgesic effect of Gonghuan Zhixue Tablet (GHZXT) on mice and to produce experimental evidence for exploiting new drug for endometrorrhagia caused by Cu-intrauterine contraceptive device (Cu IUD). METHODS: Compared with 6-aminocaproic acid and notoginseng, the effects of GHZXT on clotting and bleeding time of mice with capillary method and severed tail were investigated; and compared with aspirin, the analgesic effects of GHZXT on mice were investigated with hot plate and torsive body method. RESULTS: The clotting time of mice was remarkably shortened with a rising of the dosage of GHZXT and the difference between each therapeutic group and distilled water group was remarkable. As compared with distilled water group, the bleeding time of each dosage group of GHZXT was obviously shortened; and each dosage of GHZXT could prolong the time of pain reaction to hot plate and decrease the degree of torsive body of the mice. CONCLUSION: Pharmacological experiment has proved that GHZXT has evident hemostatic and analgesic function. PMID- 15339477 TI - [Comparison of animal models of hyperlipidemia]. AB - OBJECTIVE: To select the proper experimental animal model for research on prevention and treatment of hyperlipidemia. METHOD: Hyperlipidemia models of mouse, rat, golden hamster, guinea pig, rabbit, pigeon and quail often used in the last ten years were compared. RESULTS: Golden hamster and guinea pig models are similar to human beings in lipid metabolism and have unique superiority in experimental study, while the models of rat, mouse, pigeon and quail have significant difference as compared with human beings. Rabbit is one of the best animal models in studying new drugs for regulating lipid and treating arteriosclerosis. CONCLUSION: Factors such as the required time, indexes, the duration of making models, and the cost and feeding of animals should be considered in selecting animal models according with the experimental requirements. PMID- 15339478 TI - [Determination of catalpol in dried rehmannia root and Taohong Siwu Decoction with high performance liquid chromatography]. AB - OBJECTIVE: To determinate the catalpol contents in dried rehmannia root and Taohong Siwu Decoction containing rehmannia root with high performance liquid chromatography (HPLC). METHODS: Catalpol was separated on a YWG-C18 column using water-acetonitrile (99.4:0.6) as mobile phase and detective wavelength at 210 nm. RESULTS: The linear curve of tested catalpol concentration within the range of 0.0536-5.3600 microg/microl was ideal (n=5, r=0.999 7). The average recovery rate of the dried rehmannia root and Taohong Siwu decoction was 98.7% (RSD=0.48%) and 98.2% (RSD=1.29%) respectively. CONCLUSION: HPLC method is accurate and valuable for the quality control of Radix Rehmanniae and Taohong Siwu Decoction. PMID- 15339479 TI - [Clinical research on effects of Bushen Huoxue Decoction in treating infertility owing to ovary dysfunction]. PMID- 15339480 TI - [Effect of three-step acupuncture and cupping therapy on cervicogenic trigeminal neuralgia: a report of clinical observation]. PMID- 15339481 TI - [Clinical application of eight-magic-turtle acupuncture technique elucidated through diagrams]. PMID- 15339482 TI - [Treatment of recurrent abortion with integrated traditional Chinese and western medicine: a report of 2 cases]. PMID- 15339483 TI - [Progress of research on mechanism of salvia miltiorrhiza and its chemical ingredients against liver fibrosis]. PMID- 15339484 TI - [Progress of study on Th1/Th2 balance related to research on traditional Chinese medicine]. PMID- 15339485 TI - [Clinical observation on treatment of depression with scalp electro-acupuncture: a report of 30 cases]. PMID- 15339486 TI - [Progress of study on the regulation of endothelin and nitrix oxide with Chinese medical herbs]. PMID- 15339488 TI - [Characteristics of English translation of traditional Chinese medicine]. PMID- 15339487 TI - [Some insights into clinical detection and analysis of serum lipoprotein for studies on integrated traditional Chinese and western medicine]. AB - Dyslipoproteinemia is correlated with the occurrence and development of atherosclerosis and cardiocerebral vascular diseases. The metabolism of serum lipoprotein, however, is a complicated and ordered metabolic process. On one hand, every organ, tissue and even cell of the body participates in and affects the state of serum lipoprotein dynamic balance (SLDB) in different degrees. On the other hand, SLDB is a direct factor for maintaining the normality of whole vascular system, especially the vascular endothelium system. The serum lipoprotein dynamic imbalance, as an inducement or even etiological factor, can bring about various clinical diseases. Therefore, integrated analysis is needed to appropriately assess the state of SLDB. It is suggested that serum lipoprotein dynamic imbalance syndrome should be the key risk factor for those related diseases. Serum lipoprotein electrophoretogram can ideally express the state of SLDB and be taken as a new index for cognizing the serum lipid metabolism and the function of speen-qi to distribute Jing (Essonse) in the integrated traditional Chinese and western medicine. Practice has showed that concrete integration and bilateral development should be one of the main tasks in current researches of integrated traditional Chinese and western medicine. PMID- 15339489 TI - [Innate immunity and integration of traditional Chinese and western medicine]. PMID- 15339490 TI - [Pondering over some blunders on scientificity in diabetic researches by integrated Chinese and western medicine]. AB - Scientificity was emphasized in researches on diabetes mellitus in integrated traditional Chinese and western medicine abroad. In clinical researches, random and double-blind method, placebo control and auto-cross-over experiment were employed, and the diet, activity, drug dosage and body weight were under strict control, although such kind of researches was rare. During the past few decades, there were also some good achievements in the researches on diabetes mellitus in integrated traditional Chinese and western medicine in China, but most of the researches were duplicated work at low level. There existed some problems, such as the selection of investigated objects, control of the facts besides of the drugs, randomized grouping, blind method, auto-cross-over control, integrated research of multi-center, selection of indexes, methods of index detection, selection of complex prescription, single drug or monome, animal model, etc. The scientificity of the researches in China should be further improved. PMID- 15339491 TI - [Experience of treating severe acute respiratory syndrome with traditional Chinese medicine]. PMID- 15339492 TI - [Application of the theory of damp-heat in spleen and stomach in treatment of kidney diseases]. AB - There exists close relationship between the damp-heat and the occurrence, development, treatment and prognosis of kidney diseases. The occurrence of damp heat is based on water and wetness, which may come from outside or may be caused by internal damage. The water and wetness can change into heat if they are stored in the body for a long period, and the combination of water and wetness and heat may cause damp-heat syndrome. The function of spleen and stomach plays the most important role in the water metabolism of sanjiao. Kidney diseases are mainly caused by damp-heat of spleen and stomach resulted from internal damage of spleen and stomach, and symptoms of damp-heat of middle-jiao are commonly seen in most patients of chronic kidney diseases. During the development from renal function damage to renal failure, the remaining of damp-heat in san-jiao can affect the function of spleen and stomach for transportation, transformation and digestion, resulting in the symptoms of dysfunction of spleen and stomach. Regulating the function of spleen and stomach is one of the most important therapeutic methods for kidney diseases. PMID- 15339493 TI - [Inspiration to basic researches of traditional Chinese medicine from the 2001 Nobel Prize]. AB - The contributions of the three winners (L Hartwell, RT Hunt and PM Nurse) of the 2001 Nobel Prize for physiology or medicine revealed the mystical veil of cell cycle control. It was of far-reaching significance for exploring new method for cancer treatment. It will also give a good deal of enlightenment to the basic research of traditional Chinese medicine. Their understanding about the cause and development of cancers changed from the static view to dynamic dialectical analysis, from simplex study to comprehensive analysis; they stressed regulation, instead of killing in the treatment of cancer; and they thought that the numerous factors driving the normal process of the cell cycle could be summarized as positive and negative factors. These opinions were similar to some theories of traditional Chinese medicine, such as treatment based on syndrome differentiation, integrative treatment, and keeping the balance between yin and yang, and established a connection between traditional Chinese medicine and the western medicine, which would further widen the research on compound prescriptions of Chinese herbs. PMID- 15339494 TI - [Pathogenesis of insulin resistance and prevention and treatment strategy of integrated traditional Chinese and western medicine]. AB - Insulin resistance is the common pathophysiologic basis of multiple metabolic diseases, but the exact mechanism of insulin resistance is still unclear presently. In the past few years, studies on the pathogenesis of insulin resistance, in which the endocrine function of adipocyte and inflammation reaction participate, as well as the insulin-sensitizing drugs have become the new hotspot. The exploration on pathogenesis and treatment of insulin resistance in traditional Chinese medicine has established a foundation for the prevention and treatment of insulin resistance in integrated traditional Chinese and western medicine. PMID- 15339495 TI - [Effect of kidney-warming and astringent therapy on plasma endothelin and interleukin-2 receptor in patients with nephrotic syndrome]. AB - OBJECTIVE: To explore the effect and mechanism of kidney-warming and astringent therapy in treating nephrotic syndrome patients with deficiency of spleen and kidney yang and overflow of water, and to observe the change of plasma endothelin and interleukin-2 receptor after treatment. METHODS: Forty-four patients were randomly divided into conventional steroid treated group (control group, 20 cases) and conventional steroid plus kidney-warming and astringent therapy treated group (treatment group, 24 cases). The levels of plasma endothelin (ET) and soluble interleukin-2 receptor (sIL-2) were observed. RESULTS: Before treatment, plasma ET and sIL-2 in the patients were significantly higher than those in healthy people (P<0.01). After treatment, the ET and sIL-2 levels were obviously improved in both treated groups (P<0.05) and the improvement in the treatment group was more obvious. The difference between the two treated groups after treatment was significant (P<0.05). CONCLUSION: Conventional treatment plus kidney-warming and astringent therapy can effectively improve the levels of plasma ET and sIL-2 in treating nephrotic syndrome patients with deficiency of spleen and kidney yang and overflow of water, and hence alleviate the damage of renal tissue. PMID- 15339496 TI - [A study on detecting specific antibodies of hemorrhagic fever with renal syndrome and treatment with integrated traditional Chinese and western medicine]. AB - OBJECTIVE: To explore a simple speedy specific and sensitive method to detect specific IgM (sIgM) and IgG (sIgG) antibodies of hemorrhagic fever with renal syndrome (HFRS),and to study the therapeutic effects of integrated traditional Chinese and western medicine on HFRS. METHODS: The serum of 559 patients with HFRS were tested with colloidal gold immuno-dot assay (CGIDA) for sIgM and sIgG antibodies and compared with enzyme linked immunosorbent assay (ELISA) or indirect fluorescent antibody test (IFAT). One hundred and one patients with HFRS were randomized into treatment group (n=50),treated with Kuhuang Injection, Shenmai Injection and Huangqi Liquid) and control group (n=51),treated with Ribarvirin and Ganlixin Injection). RESULTS: The positive rate of sIgM detected with CGIDA was 70.8% and the positive rate of sIgG detected with CGIDA was 87.5%. The days for fever decline, symptoms alleviation and sign relief between the treatment group and control group were similar (P>0.05). The days for recovery of kidney function in the control group was less than that in the treatment group (P<0.01). The rate of crossing shock stage in the treatment group was higher than that of the control group (P<0.01). CONCLUSION: CGIDA was more simple, speedy, specific and sensitive than ELISA or IFAT in detecting the sIgM or sIgG antibodies in serum of patients with HFRS. Although the sensitivity of CGIDA was lower than that of ELISA the CGIDA had no false positive reaction the sensitivity of CGIDA was higher than that of IFAT on detecting IgG. The effect of the treatment group was similar to that of the control group. But the crossing shock stage rate in the treatment group was higher than that of the control group while the control group was better than the treatment group in recovering the kidney function. PMID- 15339497 TI - [Effect of Jianwei Yuyang Granules on expression of epidermal growth factor receptor in gastric mucosa of gastric ulcer patients]. AB - OBJECTIVE: To observe the effect of Jianwei Yuyang Granules on the expression of epidermal growth factor receptor (EGFR) in the gastric mucosa of gastric ulcer patients. METHODS: Sixty gastric ulcer patients (final diagnosis by gastroscope) were randomly divided into Jianwei Yuyang Granules treated group (JWYY group, n=30) and western medicine treated group (control group, n=30). Ten patients without gastric mucosa lesion were treated as normal control group. The expression of EGFR in the mucosa was tested by immunohistochemistry and RT-PCR. RESULTS: Expression of EGFR and EGFR mRNA was not observed in patients without gastric mucosa lesion, and slight expression was observed in the edge of ulcer region of the gastric ulcer patients. The expression of EGFR and EGFR mRNA was increased in JWYY group and control group. CONCLUSION: Jianwei Yuyang Granules enhances the expression of EGFR in the mucosa of gastric ulcer patients to prevent peptic ulcer recurrence. PMID- 15339498 TI - [A study on relationship of purple tongue and its regional blood flow]. AB - OBJECTIVE: To study the relationship of purple tongue and its regional blood flow. METHODS: Laser-Doppler flow meter was employed to test the volume and velocity of blood flow at different sites of purple tongue (56 cases) and normal tongue (54 cases), and the test results were analyzed. RESULTS: The volume of blood flow of purple tongue was significantly lower than that of the normal tongue (P<0.01), while the velocity was significantly higher (P<0.01). The volume and velocity of blood flow at different sites of the tongue were similar without significant difference. CONCLUSION: The volume of blood flow of purple tongue is decreased and the velocity increased. Purple tongue may be caused by dysfunction of the microcirculation of the tongue. PMID- 15339499 TI - [Effect of curcumin on extracellular matrix accumulation in the glomeruli in nephrotoxic sera nephritis rats]. AB - OBJECTIVE: To observe whether curcumin could inhibit the accumulation of the collagen IV and fibronectin in the glomeruli in nephrotoxi sera nephritis rats. METHODS: Seventy-two healthy male Sprague-Dawley rats were divided into three groups, with 24 animals in each group. For normal control group, normal saline (0.5 ml/d) was injected through intra-caudal-vein for two days, and at the same time normal saline (0.5 ml/kg) was also daily administered intraperitoneally. For nephrotoxic sera nephritis group, nephrotoxic sera (0.5 ml/d) was injected through the tail vein for two days and dimethyl sulfoxide (0.5 ml/kg) was given intraperitoneally daily. For curcumin group, nephrotoxic sera was injected as above and meanwhile curcumin (50 mg.kg(-1).d(-1)) was administered intraperitoneally every day. Six rats in each group were killed on the 3rd, 7th, 14th and 28th day. Their renal tissue was fixed in 10% formalin for examining the expression of collagen IV and fibronectin. RESULTS: Minimal staining of collagen IV and fibronectin was detected in the basement membrane of normal control rats glomeruli. In the nephrotoxic sera nephritis rats and curcumin treated nephrotoxic sera nephritis rats, the accumulation of collagen IV and fibronectin was increased progressively, with significant difference in the accumulation of collagen IV (P<0.01) between these two groups at the same time points, while the significant difference in fibronectin accumulation (P<0.05) appeared only after the 7th days. CONCLUSION: Curcumin can reduce the accumulation of collagen IV and fibronectin in the glomeruli. Hence we postulated that curcumin might have beneficial effect for retarding glomerulosclerosis. PMID- 15339500 TI - [Effects of Xinmailong Xiangdan and Shenmai Injection on early kidney damage induced by toxin of grass carp bile]. AB - OBJECTIVE: To observe the effects of Xinmailong (XML), Xiangdan (XD) and Shenmai (SM) Injection on early kidney damage induced by toxin of grass carp bile (GCB) and provide experimental evidence for Chinese medicine in treating toxication of GCB. METHODS: GCB (6ml/kg) was administered orally to adult SD rats to induce the model of GCB intoxication and a single intraperitoneal injection of XML, XD or SM was administered 10 min later. RESULTS: It was found that the levels of blood creatinine and blood urea nitrogen were decreased and the creatinine clearance rate was increased by XML, XD or SM. Degenerated epithelial cells of proximal convoluted tubules and glomeruli containing red blood cells in saccular cavities were decreased as compared with the normal group (P<0.01). CONCLUSION: The Chinese medicine XML, XD and SM Injection have therapeutic effects on early kidney damage induced by toxin of GCB. PMID- 15339501 TI - [Treatment of IgA nephropathy based on syndrome differentiation: a report of 20 cases]. PMID- 15339502 TI - [Changes of SOD, GSH and MDA in renal tissues of diabetic hyperlipidemia rats]. AB - OBJECTIVE: To study the changes of SOD, GSH and MDA in the renal tissues of diabetes rat models through the comparison of their metabolic features. METHODS: Two types of diabetes rat models were established by feeding high-lipid food along with small dose injecting of streptozotocin (STZ) (model group 1) and repeated small dose injecting of STZ with complete Freund's adjuvant (CFA) (model group 2). And changes of the following were observed: body weight, urine volume, blood sugar, blood lipid, serum insulin, insulin sensitive index (ISI),superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) of the renal tissues. RESULTS: The body weight of the two model groups of rats decreased, while the urine volume and the blood sugar increased predominantly. These indexes of the two model groups were significantly different from those of the normal group (P<0.05). The renal tissue MDA level increased while the SOD and GSH level decreased in both model group 1 and model group 2, but the changes in model group 1 were more obvious than those in the model group 2. The serum insulin level which decreased in model group 2 did not decrease significantly in model group 1, but the ISI in model group 1 decreased more significantly than that in model group 2 (P<0.05). CONCLUSION: The antioxidase activity decreased in renal tissues of diabetes rats and hyperlipidemia was the main cause of the corresponding oxidations. PMID- 15339503 TI - [Effect of tetromethylpyrazine and aminoguanidine on expression of vascular endothelial growth factor in kidney of diabetic rats]. AB - OBJECTIVE: To investigate the therapeutic mechanism of tetromethylpyrazine and aminoguanidine on diabetic nephropathy. METHODS: Diabetic rats were induced by streptozotocin. They were divided into 5 groups: normal control group (group C), untreated diabetic group (group DM), tetromethylpyrazine treated group (group TMP), aminoguanidine treated group (group AG) and tetromethylpyrazine and aminoguanidine treated group (group TMP+AG). The expression of vascular endothelial growth factor (VEGF) in renal cortex of the rats in each group was observed by immunohistochemical staining after 12 weeks. RESULTS: The expression of VEGF in renal cortex of the rats in group TMP+AG and group C was alike. The expression of VEGF in renal cortex of group TMP and group AG decreased significantly as compared with that of group C, but was still above normal level. CONCLUSION: The therapeutic mechanism of tetromethylpyrazine and aminoguanidine on diabetic nephropathy may be inhibiting the over-expression of VEGF in kidney of diabetic rats. PMID- 15339504 TI - [Effect of recombinant adenoviruses with CD/TK fusion suicide gene on human hepatocellular carcinoma cells]. AB - OBJECTIVE: To investigate gene-therapy for human hepatocellular carcinoma with adenovirus vectors by double suicide gene CD/TK. METHODS: Double suicide gene CD/TK was liberated from eukaryotic vectors pCEA-CD/TK and subcloned into shuttle vectors, and the transfer plasmid pAdtrack-CMV-CD/TK was formed after linearizing with Pac 1. It was recombinated with pAdeasy-1 in bacteria BJ5183. The identified adenovirus plasmid was digested by Pac1 and was transfected into 293 cells to pack the adenoviruses. After PCR determination, its titre was measured, and the infection rate and efficacy were tested in human hepatocellular carcinoma cells. RESULTS: pAdtrack-CMV-CD/TK and pAd-CD/TK were tested by endonuclease digestion. Ad-CD/TK was produced in 293 cells, and the human hepatocellular carcinoma cells (SMMC7721) infected by Ad-CD/TK were killed after 5-FC was used, and bystander effects were observed. CONCLUSION: Recombinant adenoviruses with CD/TK fusion suicide gene have a high infection rate and efficacy for human hepatocellular carcinoma cells. PMID- 15339505 TI - [Effect of reducing caloric intake on mice transplanted with S180 ascitic cancer]. AB - OBJECTIVE: To examine the effect of different reduced caloric intake on mice transplanted with S180 ascitic tumor. METHODS: The institute for cancer research (ICR) mice were randomly divided into control group, 3.0 standard feed (SF) group, 2.0 SF group and 1.3 SF group. The mice in control group were fed enough (about 5 g/d) dietary intake, while the amounts of dietary intake in the latter three groups were scaled down in the proportion of 65%, which were 3.0 g, 2.0 g and 1.3 g standard feed respectively. Meanwhile the essential vitamins were added to the latter three groups to keep the amount of intake the same as that of the control's. RESULTS: For most of the mice, the caloric intake obviously prolonged the mean survival days and improved the life quality was 7.14 kcal/d, and the fasting blood glucose level was 2-3 mmol/L. CONCLUSION: Properly reduced caloric intake and keeping lower blood glucose level is beneficial to prolonging the survival time of mice transplanted with S180 ascitic cancer. PMID- 15339506 TI - [Experimental study on Qi-replenishing and Yin-nourishing in inhibiting tumor metastasis]. AB - OBJECTIVE: To study the effect of Qi-replenishing and Yin-nourishing in inhibiting tumor metastasis. METHODS: Fifty mice were randomly divided into replenishing Qi group, replenishing Yin group, replenishing Qi and Yin group, chemotherapy group and control group. Models of Lewis lung cancer and B(16) melanoma were observed in vivo. The tumor weight inhibiting rate, metastatic lung tumor, concentrations of t-PA and TXB(2) in tumor tissue and IL-2 and TNF-alpha in spleen were observed. RESULTS: The tumor weight inhibiting rates of replenishing Qi and Yin group for Lewis lung cancer and B16 melanoma were both 42.9%. Replenishing Qi and Yin could decrease the contents of TXB(2) and 6-keto PGF(1alpha), and increase the levels of IL-2 and TNF-alpha. CONCLUSION: Replenishing Qi and Yin can promote the healthy energy to inhibiting tumor metastasis. PMID- 15339507 TI - [Application of traditional Chinese medicine in treating acute infection of throat]. PMID- 15339508 TI - [Effects of kidney-tonifying Chinese herbal medicine on hypothalamic-pituitary gonadotrophic function]. AB - OBJECTIVE: To observe the effects of kidney-tonifying Chinese herbal medicine on the synthesis and secretion of gonadotropin releasing hormone (GnRH) and the related neurotransmitters and neuropeptides, and to explore the mechanism of the regulative effect of Chinese herbal medicine on the hypothalamic-pituitary gonadotrophic function. METHODS: Female Sprague-Dawley rats during the period of normal adolescent initiation (160-180 g, 1.5 months) were randomly divided into three groups. The control group was fed with normal saline and the two experimental groups were fed with Chinese herbal medicine for nourishing yin to reduce fire (Zi Yin Xie Huo, ZYXH) or nourishing kidney to replenish essence (Yi Shen Tian Jing, YSTJ) respectively. The dosage was 5 ml/d for 30 days by gastric gavage. Integrated optic densities of the GnRH and neuropeptide Y (NPY) in medial preoptic area (MPOA), arcuate nucleus (ARC) and median eminence (ME) of hypothalamus were determined by immunohistochemistry method and image processing. The content of GnRH in homogenate of hypothalamus preoptic area was determined by radioimmunoassay (RIA). The releasing amount of monoamine neurotransmitters from medial basal hypothalamus (MBH) was determined by brain slices incubation and high pressure liquid chromatography (HPLC). The releasing amount of GnRH and NPY from POA was determined by push-pull perfusion and RIA, and the releasing amount of monoamine neurotransmitters from this site was determined by HPLC. RESULTS: ZYXH could inhibit the synthesis and secretion of GnRH from periodic and tonic secretory centers of GnRH, while YSTJ could stimulate the synthesis and secretion of GnRH from the both secretory centers of GnRH. ZYXH could inhibit the activity of GnRH neurons via diminishing the releasing of norepinephrine (NE) from tonic secretory center of GnRH, increasing the releasing of dopamine (DA) in periodic secretory center of GnRH and increasing the synthesis and releasing of NPY from the both secretory centers of GnRH, it hence inhibited the hypothalamic-pituitary gonadotrophic function. YSTJ could stimulate the activity of GnRH neurons via diminishing the synthesis and releasing of NPY from tonic secretory center of GnRH, it hence promoted the hypothalamic-pituitary gonadotrophic function. CONCLUSION: Chinese herbal medicine for tonifying the kidney could modulate hypothalamic-pituitary gonadotrophic function via regulating the synthesis and secretion of GnRH and the related neurotransmitters (NE, DA) and neuropeptides (NPY). PMID- 15339509 TI - [Genetic identification of internal transcribed spacers sequence in rDNA of Artemisis iwayomogi Kitam. and other two Artemisia species]. AB - OBJECTIVE: To make an useful identification method for the molecule of DNA on 3 herbs of Artemisia genus and compare the differences of the genes of Korean and Chinese species of Artemisia. METHODS: Sequence of 3 herbs (Artemisia sacrorum Ledeb., Artemisia iwayomogi Kitam. and Artemisia capillaris Thunb.) was determined by PCR sequence system. DNA was extracted from rDNA/ITS (internal transcribed spacers) and 5.8 s. The analysis was based on the amplification through DNA sequence system. RESULTS: There were profound differences between the Korean Artemisia and Artemisia sacrorum L. These 2 herbs had a difference in the PCR amplifications of the agarose gel electrophoresis. There was a slight difference in the analysis of the DNA sequence system, and the substitution percentage for ITS gene fragments sequence was 3.96%. CONCLUSION: Analytic identification method on sequence system of ITS in rDNA is effective for these 3 herbs. PMID- 15339510 TI - [Measurement of zinc in Mongolia patent drug Zhuangxiyin Powder with pulse stripping voltammetry]. AB - OBJECTIVE: To determine the content of zinc in Mongolia patent drug Zhuangxiyin Powder. METHODS: Differential pulse stripping voltammetry was employed for measurement of zinc. RESULTS: The zinc content in three samples of the drug was (493+/-11.95)microg/g, (526+/-13.74)microg/g and (554+/-9.84) microg/g respectively, and the relative standard deviation (RSD) was 2.42%, 2.61% and 1.78% respectively. CONCLUSION: The content of zinc in Zhuangxiyin Powder of daily dosage is higher than the needed daily intake of healthy people. PMID- 15339511 TI - [Treatment of diabetic skin ulcers by invigorating Qi and dispersing stagnation: a report of 38 cases]. PMID- 15339512 TI - [Treatment of chronic atrophic gastritis based on syndrome differentiation: a report of 52 cases]. PMID- 15339513 TI - [An overview of medicinal plant resource for decreasing blood glucose]. PMID- 15339514 TI - [Recent clinical researches on diabetic gangrene in integrated traditional Chinese and western medicine]. PMID- 15339515 TI - [Research progress in Fagopyrum cymosum for treating cancers]. PMID- 15339516 TI - [Traditional Chinese medicine in treatment of Parkinson's disease]. AB - Parkinson's disease is a commonly encountered central nervous retrograde affection in elder persons. According to the theories of traditional Chinese medicine, Parkinson's disease is characterized by deficiency in the Ben (root) and excess in the Biao (branch). The Ben (root) is insufficiency of liver and kidney and deficiency of qi and blood; and the Biao (branch) is wind, fire, phlegm and stasis. Good therapeutic effects have been obtained by treatment based on syndrome differentiation, treatment with specific prescriptions and acupuncture. The further study on literatures, standardized criterion of symptoms differentiation and therapeutic effect determination as well as the screening and further research on the effective prescriptions and herbs should be paid more attention, meanwhile, the integration of herbs and acupuncture will be conducive to raise the therapeutic effect. PMID- 15339517 TI - [Effects of language and translation on spreading of traditional Chinese medicine to the west]. PMID- 15339518 TI - [Discussion on definition of "traditional Chinese medicine and western medicine integration" and "integrated traditional Chinese and western medicine"]. AB - OBJECTIVE: To discuss the definition of "traditional Chinese medicine and western medicine integration" and "integrated traditional Chinese and western medicine". METHODS: Logic methods. RESULTS: It was found that there were a lot of problems on the definition of "traditional Chinese medicine and western medicine integration" and "integrated traditional Chinese and western medicine". CONCLUSION: The understanding of the concepts of "traditional Chinese medicine and western medicine integration" and "integrated traditional Chinese and western medicine" was improved after the extensive discussion. PMID- 15339519 TI - [Establishment and development of clinical theory for integration of traditional Chinese and western medicine]. AB - This paper explored the clinical thinking way in treating disease by integration of traditional Chinese and western medicine(ITCWM). In accordance with the challenge and problems the subject of ITCWM faced, the standard of diagnosis, treatment and evaluation of therapeutic response should be established. The combination of symptoms-differentiation with special prescription/drug for special disease to increase the accuracy of treatment is the developing direction for clinical medicine and ITCWM. Absorption and utilization of modern science and technique is the prerequisite for breaking through in ITCWM theory. The key of combining two different medical theories is to look for link joint. PMID- 15339520 TI - [History and causes of acupuncture advancing towards the world]. AB - Acupuncture therapy is one of the original innovations in traditional Chinese medicine. The study of acupuncture has gradually advanced towards the world since 1970s. In these years the most important events might be the influence of acupuncture anesthesia, the attention of the World Health Organization, the establishment and development of the World Federation of Acupuncture and Moxibustion Societies, and the consensus development conference on acupuncture held by the National Institutes of Health in America, and so on. The reasons why acupuncture is accepted by the world are both the convinced curative effects and the scientific foundations, due to the great efforts of medical stuff in China guided by the principle of integration of traditional Chinese and western medicine. This article poses several proposals about further extending the influence of acupuncture. In brief, acupuncture research is one of the few fields that authentically impact the western scientific technology. It is of very important responsibility in the new century to impulse the acupuncture study to further progress and expand the international influence of acupuncture. PMID- 15339521 TI - [Experience of Professor Tang Han-Jun in treating obstinate and severe surgical diseases]. PMID- 15339522 TI - [Recognition on toxicity of traditional Chinese drugs]. AB - Our ancestor found that some of the Chinese herbal drugs were toxic during their clinical practice. They graded the toxicity of Chinese herbal drugs into three degrees and thought that the term "toxicity of the Chinese herbal drugs" could be used in a broad and a narrow sense. In modern times, toxic components of the Chinese herbal drugs and their toxic mechanisms, especially the toxicity on the kidney, were further revealed. The factors that affect the toxicity of the Chinese herbal drugs include the species, preparation, dose and environment. To prevent the toxicity of the Chinese herbal drugs, we must strengthen the management of the Chinese herbal drugs. On the other hand, doctors' recognition of the toxicity of Chinese herbal drugs should be enhanced. At the same time, patients should be told to decoct and take Chinese herbal drugs correctly. PMID- 15339523 TI - [Pathogenic factors of blood circulation disturbance in lumbar intervertebral disc herniation and mechanism of Tuina manipulation in promoting circulation]. AB - Tuina manipulation has long been used in the treatment of lumbar intervertebral disc herniation, although its therapeutic mechanisms remain uncertain. The specific characteristic syndrome of this disease is lumbocrural pain in varying degrees, which is due to mechanical compression, aseptic inflammation, blood circulation disturbance and dystrophy. These factors can act on the involved nerve roots and give rise to nerve conduction disturbance. Since the blood circulation disturbance and nerve roots dystrophy are the most important pathogenic aspects of this disease, the therapeutic effect of Tuina manipulation on lumbar intervertebral disc herniation is related to them directly or indirectly. PMID- 15339524 TI - [Treatment of temporomandibular joint disorder by traditional Chinese medicine]. PMID- 15339525 TI - [Influence of cervical curvature change on the nerve root and vertebral artery of patients with cervical spondylosis and clinical observation of manipulation treatment]. AB - OBJECTIVE: To observe the influence of the change of cervical curvature on the nerve root and the vertebral artery of the patients with cervical spondylosis and the curative effect of therapeutic manipulations. METHODS: Sixty cases of cervical spondylosis with the change of cervical curvature, 60 cases of cervical spondylosis without the change of cervical curvature and 60 normal people were chosen. They were clinically observed and the cervical spondylosis was treated by manipulations and the data was processed according to the principle of statistics. RESULTS: The change of cervical curvature affected the nerve root and the vertebral artery and therapeutic manipulations produced a satisfactory curative effect. CONCLUSION: Cervical spondylosis with change of cervical curvature has special characteristics in the etiology, course and state of the disease. The change of cervical curvature produces an effect on the nerve root and the vertebral artery. Selected therapeutic manipulations produce a better curative effect. PMID- 15339526 TI - [An transcranial Doppler ultrasonography and X-ray study of cervical vertigo patients treated by manipulation in supine position]. AB - OBJECTIVE: To observe the transcranial Doppler ultrasonography and X-ray changes of cervical vertigo patients caused by cervical instability before and after the manipulation treatment in supine position. METHODS: Sixteen cervical vertigo patients who underwent supine manipulation treatment were reviewed retrospectively. After the treatment, the changes of transcranial Doppler ultrasonography of cervical vertebral artery and X-ray of cervical spine were observed. RESULTS: Before the treatment, the mean blood flow velocity of vertebral artery was (48.6+/-3.1) cm/s, much higher than that of the normal group (P<0.05), the means of the horizontal displacement of C3,4 and C4,5 were (3.18+/ 0.42) mm and (3.46+/-0.26) mm respectively. After the treatment, the blood flow velocity and the C3,4 horizontal displacement decreased significantly (P<0.05). The effective rate of the manipulation treatment was 100%. CONCLUSION: The manipulation treatment is an effective method for cervical vertigo patients caused by cervical instability, and the effect is correlated with the improvement of the blood flow of cervical artery and the restoration of cervical stability. PMID- 15339527 TI - [Retrospective study of the relationship of height difference and bone density of postmenopausal women]. AB - OBJECTIVE: To analyze the relationship of the height difference and bone density (BD) of premenopausal and postmenopausal women. METHODS: The height values of 191 premenopausal and postmenopausal women were recorded, and the BD values of lumbar vertebrae and hip were detected by double energy X-ray BD detector. RESULTS: The lower the height of the postmenopausal women, the less the BD value. The BD of lumbar vertebrae dropped 0.025 5 g/cm(2) with each 2 cm of the shortened height, and the BD of hip joint dropped 0.029 2 g/cm(2). The shortened value in postmenopausal women with osteoporosis was statistically greater than that in postmenopausal women without osteoporosis. CONCLUSION: The BD of the postmenopausal women can be estimated by the calculation of their shortened height value. PMID- 15339528 TI - [Clinical analysis of 82 cases of ectopic pregnancy treated by methotrexate combined with traditional Chinese recipe]. PMID- 15339529 TI - [Clinic research on heroin de-addiction effects of acupuncture and its potentiality of preventing relapse]. AB - OBJECTIVE: To compare the effects of de-addiction with the therapy of acupuncture, acupuncture plus opium, opium plus buprenorphine and opium plus Han's instrument for de-addiction and to study the effects of the four therapeutic methods on the protracted withdrawal syndrome and craving. METHODS: The effects of de-addiction were assessed with the opiate withdrawal scale and the craving degree with visual analogue scale (VAS). RESULTS: The dominance of acupuncture treatment for withdrawal syndrome appeared to be after the 6th day, and the dominance for controlling craving showed after the 8th day, moreover, there were little side effects. CONCLUSION: Acupuncture treatment had the potentiality of preventing relapse and could be used for treating the protracted withdrawal syndrome and psychic dependence during the period between the stages of abstinence and rehabilitation. PMID- 15339530 TI - [Regulation effects of Gegen Recipe and Guizhi Recipe on expression of IL-1beta, iNOS, TNFalpha, TGFbeta mRNA in degenerated cervical intervertebral disk in rabbits]. AB - OBJECTIVE: To study the effects of Gegen Recipe and Guizhi Recipe regulating the expression of IL-1beta, iNOS, TNFalpha and TGFbeta mRNA in degenerated intervertebral disk caused by stimulation of wind, cold and damp. METHODS: Thirty six rabbits were randomly separated into six groups, viz. control group, stimulation by wind, cold and damp group, head in low-location group, compound model group (head in low-location plus stimulation of wind, cold and damp), Gegen Recipe group and Guizhi Recipe group. The gene expression of IL-1beta, iNOS, TNFalpha and TGFbeta mRNA in degenerated cervical intervertebral disks were assayed by RT-PCR. RESULTS: Compared with the control group, IL-1beta mRNA and iNOS mRNA expression were up-regulated in each model group(P<0.01). Both Gegen Recipe and Guizhi Recipe down-regulated IL-1beta mRNA, iNOS mRNA expression, which had significant differences compared with the stimulation model group(P<0.01 or <0.05). TNFalpha mRNA expression was significantly enhanced in the compound model group compared with the control group (P<0.01). There was no significant difference between the head in low-location group and the stimulation model group. Gegen Recipe down-regulated TNFalpha mRNA expression compared with the stimulation group(P<0.01),while the effect of Guizhi Recipe was not significant. Compared with the control group, TGFbeta mRNA expression went down in each model group(P<0.01). Gegen Recipe up-regulated TGFbeta mRNA expression(P<0.01), while the effect of Guizhi Recipe on adjusting TGFbeta mRNA was not significant. CONCLUSION: Stimulation by wind, cold and damp, head in low location, and compound of both can down-regulate the expressions of IL-1beta, iNOS, TNFalpha mRNA, and up-regulate the expression of TGFbeta mRNA. These researches certify that these factors take part in the progression of intervertebral disk degeneration. IL-1beta, iNOS, TNFalpha accelerate the degeneration and TGFbeta defers the degeneration. Gegen Recipe down-regulates the expression of TNFalpha mRNA and up-regulates the expression of TGFbeta mRNA. The effect on regulating the cell factors expressions by Guizhi Recipe is lower than that of Gegen Recipe. PMID- 15339531 TI - [Effect of Yiqi Huayu Recipe on regeneration process after L5 nerve root compression in rats]. AB - OBJECTIVE: To study the effect of Yiqi Huayu Recipe on neurons in neuromuscular junction after L5 nerve root compression in rats. METHODS: By using immunohistochemistry, confocal laser scanning techniques and protein gene product 9.5 as a neuronal marker, the motor endplates were visualized with fluorescein conjugated alpha-BTX. The effect of Yiqi Huayu Recipe on nerve terminals of neuromuscular junction at 10, 20, 30, 60 days after L5 nerve root compression was observed in rat soleus muscles during regeneration. The overlap areas of terminals and endplates were measured with NIH image technique. RESULTS: As for the aggregates, sprouts, extension of nerve terminal during regeneration, the connecting speed and range of nerve terminal with endplates, as well as the reformation of neuromuscular junctions during regeneration, Yiqi Huayu Recipe group was better than that of the control group. CONCLUSION: Yiqi Huayu Recipe can promote the hyperplasia of neurons and enhance the regeneration function. It also can accelerate the neuromuscular junction reformation and shorten the process of regeneration. PMID- 15339532 TI - [Influence of electro-acupuncture of Neiguan on plasmic concentrations of NO and TNFalpha in endotoxin shock rats]. AB - OBJECTIVE: To observe the effect of electro-acupuncture of Neiguan (PC 6) on mean systemic arterial blood pressure and plasmic concentrations of NO and TNFalpha in endotoxin shock rats. METHODS: The model of endotoxin shock was induced by lipopolysaccharide (1.5 mg/kg i.v.) and D-galactosamine (100 mg/kg i.p.). Aminoguanidine (100 mg/kg i.p.) and electro-acupuncture of bilateral Neiguan (PC 6) were administered. A catheter was inserted into the right subclavian artery to record the change of blood pressure and the blood was abstracted out and centrifuged to determine the NO and TNFalpha concentrations. RESULTS: Electro acupuncture of Neiguan (PC 6) retrieved the blood pressure and reduced the plasmic NO and TNFalpha concentrations. CONCLUSION: Electro-acupuncture of Neiguan (PC 6) expresses an anti-endotoxin shock effect by repressing the plasmic NO and TNFalpha concentrations smoothly and retrieving the blood pressure stably. PMID- 15339533 TI - [Treatment of reflux esophagitis by Xuanfu Daizhe Decoction combined with pantoprazole or omeprazole: a comparative study]. PMID- 15339534 TI - [Effect of Wuling Powder on rats with renal hypertension]. AB - OBJECTIVE: To observe the therapeutic effect of Wuling Powder extract on rats with renal hypertension and to evaluate the influence of it on the volume of urine and the concentrations of Na(+), K(+), Cl(-). METHODS: Reformed Gold-blatt hypertension rat model (G-2K1C) was established. The rats were divided into 6 groups as follows: sham-operation group; model group, Wuling Powder high dosage group (80 g/kg), Wuling Powder middle dosage group (40 g/kg), Wuling Powder low dosage group (20 g/kg), and hydrochlorothiazide (HCT) group (25 mg/kg). Urine volume of the rats was measured during the experiment. Tail arterial pressure and [Na(+)], [K(+)], [Cl(-)] in serum of the rats were detected after 30 days of treatment. RESULTS: The blood pressure of the G-2K1C rats was decreased in the three Wuling Powder groups (P<0.05 or P<0.01), but higher than that of the false operation group (P<0.01), and there was no difference between each of the Wuling Powder groups and the HCT group (P>0.05). Diuretic effect of the three dosages of Wuling Powder was weaker than that of the HCT (P<0.05 or P<0.01). The effects of the three dosages of Wuling Powder and HCT on [Na(+)] and [Cl(-)] in the serum were not obviously different (P>0.05), but [K(+)] of the HCT group was significantly decreased compared with that of the false-operation group and the three Wuling Powder groups (P<0.01). CONCLUSION: Wuling Powder extract had satisfying therapeutic effects in increasing the discharge of urine, decreasing the blood pressure and keeping the balance of the serum electrolyte contents in rats with renal hypertension. PMID- 15339535 TI - [Experimental study on effects of herbs for nourishing and smoothing the liver in reversing bile lithogenicity of guinea pig]. AB - OBJECTIVE: To further probe the mechanisms of herbs for nourishing and smoothing the liver in reversing bile lithogenicity of guinea pig. METHODS: Sixty guinea pigs were divided randomly into control group (fed with normal diet, n=20), model group (fed with lithogenic diet, n=20) and treatment group (fed with lithogenic diet plus herbal medicine, n=20). After four-week feeding, the animals were sacrificed and sampled, the rates of gallstone formation in each group were estimated, and the total bile acid (TBA), total bilirubin (TBIL), conjugated bilirubin (CB), unconjugated bilirubin (UCB), and calcium ion in the bile were determined, and the different bilirubins were analyzed by HPLC. RESULTS: (1) The rate of gallstone formation was 5% in normal group, 81.25% in model group and 31.25% in treatment group (P<0.05). (2) The bile TBIL, CB, UCB and Ca(2+) were higher and the bile TBA was lower significantly in model group than that in the other two groups (P<0.05). (3) HPLC analysis revealed that MCB was higher and DCB was lower significantly in model group (P<0.01), and there were no significant differences of UCB and IPA among the three groups. (4) The percentages of MCB and UCB were much higher and the percentage of DCB was remarkably lower in model group (P<0.05). CONCLUSION: Herbs for nourishing and smoothing the liver can significantly reduce the rate of gallstone formation and has effect of reversing lithogenicity of bile in guinea pigs fed with lithogenic diet. PMID- 15339536 TI - [Effect of Chinese and western medicine integration on spinning behavior of rats with Parkinson disease]. AB - OBJECTIVE: To observe the effect of Chinese and western medicine integration on the spinning behavior of rats with Parkinson disease. METHODS: Model of the lateral Parkinson disease was made with injection of 6-hydroxydopamine (6-OHDA) into the black substance of the right side of the brain, and the model rats were treated with madopar and Chinese herbs for nourishing the liver and kidney, clearing collaterals and detoxification. The rat's spinning behavior was observed, and was compared with the normal control group, madopar group and sham operation group at the same time. RESULTS: Chinese and western medicine integration could obviously reduce the spinning circles of the rats. CONCLUSION: Chinese and western medicine integration can significantly improve the spinning behavior of the model rats. PMID- 15339537 TI - [Experience of Professor Li Guo-Heng in treating degenerative osteoarthrosis of knee joint]. PMID- 15339538 TI - [Medical value of isoflavones]. AB - Isoflavones as a kind of phytoestrogen exist in soy plant widely. With the advantages of estrogen, they can regulate endocrine function of the human body, reduce blood cholesterol, decrease the incidence of coronary heart disease, prevent osteoporosis, inhibit tumor growth, and relieve pain. This article summarized the investigations on the medical value of isoflavones in recent years. PMID- 15339539 TI - [Recent advances of research on warming therapy]. AB - Therapeutic method is an important component of traditional Chinese medicine theory. Research on warming therapy focuses on pharmacology at present. Studying the feature of nerve-body fluid-metabolism-function in cold syndrome is of importance for illustrating the nature of warming therapy and elucidating the function of medication. This review presents the research and development of warming therapy in order to provide enlightenment for the research of therapeutic method from the aspects of traditional Chinese medicine and western medicine. PMID- 15339540 TI - [History and development of spinology]. AB - Spinal disease is the injury caused by congenital malformation, degeneration, inflammation and trauma. These injuries lead to structural lesion of spinal itself, intervertebral discs, spinal cord, nerve root and the attachment, and can cause spinal and associated diseases. Spinal diseases include some frequently encountered diseases, as well as some stubborn and serious diseases. These diseases may cause pain of head and neck, shoulder and arm, waist and leg when they are mild, and partial or thorough paralysis when they are serious. In this article, the history, development, foundations and research progress of non surgical operation of spinal diseases and the developing tendency of modern spinology are introduced. PMID- 15339541 TI - [Methods of western translators for translation of traditional Chinese medicine in early days]. PMID- 15339542 TI - [Effect of Tuina on rehabilitation of physical strength for football sportswomen]. PMID- 15339543 TI - [Clinical application of explaining Ziwu Liuzhu therapy through diagrams]. PMID- 15339544 TI - [Clinical observation of 41 cases of senile ischemic heart disease treated by losarton, carvadilol and Shenmai Injection]. PMID- 15339545 TI - [Analysis on acupuncture treatment characteristics for tumor in ancient medical literature]. PMID- 15339546 TI - [Clinical observation on 44 cases of menopause syndrome treated with Liu'an Qiwei Decoction]. PMID- 15339547 TI - [Traditional Chinese medicine in prevention and treatment of liver cancer: function, status and existed problems]. PMID- 15339548 TI - [Restoring the harmony and order of the body: treatment of cancer patients according to syndrome differentiation]. AB - By reviewing the clinical work of the past 40 years and combining with the theories of traditional Chinese medicine, the author proposed that the spleen plays a key role in treating liver cancer according to syndrome differentiation. Based on the clinical and experimental research work, the author advanced a theory that the goal of the treatment of cancer according to syndrome differentiation is to restore the harmony and order of the body. Further research into this theory will be conducive to establishing a new theoretical system of integrative medicine on cancer treatment. PMID- 15339549 TI - [Treatment of blastocystis hominis protozoosis with Baitouweng Decoction: a report of two cases]. PMID- 15339550 TI - [Problems in cancer treatment and major research of integrative medicine]. AB - There exist some problems in the present cancer treatment, such as: (1) prevalence of excessive treatment; (2) lack of long-term research plan; (3) lack of reasonable comprehensive treatment; (4) lack of operative individualized treatment; (5) the superiority of traditional Chinese medicine (TCM) was undervalued and underused. Integrative medicine should play an important role in the prevention and treatment of cancer. Further research should be carried out in the following aspects: (1) enrich and improve the theoretical knowledge of TCM on cancer; (2) enhance the research on rationalization and standardization of cancer treatment program of integrative medicine; (3) enhance the research on criterion of therapeutical effect of integrative medicine in treating cancer; (4) enhance the research on prevention and treatment of postoperative recurrence and metastasis of cancer by integrative medicine. PMID- 15339551 TI - [Clinic experience in treating two cases]. PMID- 15339552 TI - [Tumor micrometastasis and its treatment with traditional Chinese medicine]. AB - This paper discussed the detection of tumor micrometastasis, and analysed the influence of chemotherapy and blood transfusion on the immunity of tumor patients. Chemotherapy and blood transfusion can increase the opportunity of tumor metastasis. By analysing the mechanism of the treatment of tumor with traditional Chinese medicine, the author put forward the new method about eliminating tumor micrometastasis with traditional Chinese medicine. PMID- 15339553 TI - [Study on decomposed recipes of traditional Chinese medicine]. AB - OBJECTIVE: To introduce and analyze the present status of research on decomposed recipes of traditional Chinese medicine (TCM). METHODS: The relevant literature on the study of the decomposed recipes of TCM in the past five years was collected and analyzed. RESULTS AND CONCLUSION: Studies on the decomposed recipes of TCM have been made from the whole body level to the organ, cell and molecular research level. The pharmacological research in vitro is the main way and quantitative changes of the chemical ingredients have been measured before and after composition. It is very important to combine the modern science and technology in exploring the composing mechanisms of TCM remedies. PMID- 15339554 TI - [A review of experimental designs of compound compatibility law of traditional Chinese medicine]. AB - The paper discussed a variety of experimental designs of compound compatibility law of traditional Chinese medicine (TCM): study of whole formula and different ingredients of formula. The latter includes study of single ingredient, study of functional ingredient group, orthogonal design, clustering analysis, homogeneous design, factorial analysis and so on. It was proposed that experimental designs of formula should be based on the theory of TCM, and combined with modern sciences. PMID- 15339555 TI - [Quantitative study on tongue color in primary liver cancer patients by analysis system for comprehensive information of tongue diagnosis]. AB - OBJECTIVE: To explore the characteristics of tongue color in patients with primary liver cancer (PLC). METHODS: Tongue color and its RGB value were analyzed quantitatively for PLC patients in different clinical periods and other cancer patients by analysis system for comprehensive information of tongue diagnosis. RESULTS: The rate of blue and purple tongue was higher (P<0.05) and all the values of RGB were lower (P<0.01) in PLC patients compared with other cancer patients. In different clinical periods, the rate of blue and purple tongue in stage III was the highest (P<0.05). CONCLUSION: The blue and purple tongue is one of the most important tongue characteristics of PLC patients. PMID- 15339556 TI - [Clinical observation of cinobufacini injection used to treat moderate and advanced primary liver cancer]. AB - OBJECTIVE: To observe the clinical effect of cinobufacini injection in treating moderate and advanced primary liver cancer (PLC). METHODS: One hundred patients with moderate and advanced PLC were randomly divided into cino-treated group (50 patients) and control group (50 patients). The quality of life, tumor size, some changes of laboratory tests, and survival time were observed. RESULTS: The progressive rate of cino-treated group (18%) was lower than that of the control group (32%). The quality of life of the cino-treated group (80%) was better than that of the control group (72%), but without statistical significance. The survival rate of >12 months of the cino-treated group (30%) was higher than that of the control group (18%). The patients' liver function such as serum total bilirubin and ALT decreased obviously in the cino-treated group while increased a lot in the control group. The level of AFP increased after treatment with statistical significance in the control group while there was no statistical significance in the cino-treated group. CONCLUSION: Cinobufacini injection can not only inhibit the proliferation of cancer, but also protect liver function, improve quality of life and prolong survival time. PMID- 15339557 TI - [Clinical evaluation of integration of transcatheter arterial chemoembolization and traditional Chinese medicine in treating metastatic liver cancer]. AB - OBJECTIVE: To evaluate the treatment effect, quality of life and side-effect of transcatheter arterial chemoembolization (TACE) and traditional Chinese medicine (TCM) in treating metastatic liver cancer. METHODS: Thirty-nine cases of colon metastatic liver cancer were randomly divided into two groups. Both TACE and TCM were used in the treatment group, while only TACE was used in the control group. The drug used in TACE included floxuridine, pirarubicin, cisplatin, and the herbs for strengthening the spleen and regulating Qi were used in TCM. RESULTS: The response rate in the treatment group was 30% (45% including minor remission patients), and the median survival time was 18.6 months. While in the control group the response rate was 15.8% (36.8% including minor remission patients), and the median survival time was 14.3 months. The 1-, 2-, 3- year survival rates of treatment group and the control group were 70.2%, 40.3%, 13.0% and 68.7%, 29.5%, 10.3% respectively. There were fewer other organ metastases in the treatment group. The score from the EORTC quality of life questionnaire QLQ-C30 in treatment group was higher than that in the control group. CONCLUSION: Integration of TACE and TCM in treating colon metastatic liver cancer has better results. PMID- 15339558 TI - [Clinical study on treatment of patients with upper digestive tract malignant tumors of middle and late stage with Ginkgo biloba exocarp polysaccharides capsule preparation]. AB - OBJECTIVE: To observe the clinical efficacy of Ginkgo biloba exocarp polysaccharides (GBEP) capsule preparation in treating upper digestive tract malignant tumors of middle and late stage. METHODS: Eighty-six patients of the upper digestive tract malignant tumors were treated with GBEP capsule preparation taken orally. The clinical symptoms and the qualities of life of the patients with single GBEP and combined with operation, radiotherapy or intervention chemotherapy were observed. The tumor size was measured by electronic gastroscope before and after treatment with single GBEP. Objective response rate (RR) of the tumor was calculated. The survival period of patient was observed. The changes of blood routine examination in the patients treated with radiotherapy were observed. RESULTS: GBEP preparation could markedly improve the patients'clinical symptoms. Karnofsky scoring of the patients markedly increased after treatment. There were 2 CR (complete response, 6.3%), 22 PR (partial response, 68.8%)and 5 SD (stable disease, 15.6%) of 32 cases with single GBEP preparation. The survival periods of the 32 cases were markedly prolonged. The preparation could relieve the inhibited hematopietic function and the weight loss due to radiotherapy. CONCLUSION: GBEP capsule preparation has some definite therapeutic effects on upper digestive tract malignant tumors of middle and late stage. PMID- 15339559 TI - [Clinical observation on treatment for postoperative metastasis of gastric cancer by Jianpi Huoxue Jiedu prescription]. AB - OBJECTIVE: To study the effect of Jianpi Huoxue Jiedu (JPHXJD) prescription on prevention of the postoperative metastasis of gastric cancer and to observe the changes of immunity and whole blood viscosity after treatment. METHODS: JPHXJD prescription and chemotherapy were administered to the treated group for 12 successive months, while only chemotherapy was given to the control group. RESULTS: The effective rate of anti-metastasis in the treated group was obviously better than that in the control group. In the aspects of enhancing Karnofsky scores, reducing whole blood viscosity and improving immunity after treatment, the treated group was better than the control group (P<0.05). CONCLUSION: JPHXJD prescription combined with chemotherapy shows apparent effect on anti-metastasis. It can improve the quality of life of the patients, lower the whole blood viscosity and improve the immunity function. PMID- 15339560 TI - [Prognostic analysis of advanced non small cell lung cancer treated by sequential chemo-radiation therapy combined with traditional Chinese medicine: a report of 54 cases]. AB - OBJECTIVE: To evaluate the influence of fewer courses and prolonged intervals of chemotherapy on survival rate of advanced non small cell lung cancer (NSCLC) patients treated by sequential chemo-radiation therapy combined with traditional Chinese medicine (TCM). METHODS: From Jan. 2000 to Dec. 2001, 54 untreated advanced NSCLC patients (2 stage IIIa, 18 stage IIIb, 34 stage IV) were treated by sequential chemo-radiation therapy combined with TCM. The courses of chemotherapy were reduced and the intervals of chemotherapy were longer than that of the standard regimen. The efficacy and survival rate were documented and the prognostic factors were analyzed. RESULTS: Complete remission (CR) was observed in 1 case and partial remission (PR) in 20 cases. The overall objective response rate was 40.4%. Median survival was 15.3 months, 1-, 2- and 3-year survival rate were 53.7%, 28.9% and 9.6% respectively. The median survival of stage III and IV were 21.8 months and 12.5 months respectively. The 1-, 2-, and 3-year survival rates of stage III were 65.0%, 49.5%, 24.7% and that of stage IV were 47.0%, 23.3%, 0%, respectively. The quality of life was improved in most of the patients. Cox's proportional hazards regression showed that improved quality of life and treatment of TCM were the significant prognostic factors of overall survival. CONCLUSION: Chemotherapy and radiotherapy combined with TCM is beneficial to extending the interval of chemotherapy, improving the quality of life, and increasing the survival rate of advanced NSCLC patients. PMID- 15339561 TI - [Correlation between TXB2, 6-Keto-PGF1alpha and liver metastasis in rats model with blood stasis]. AB - OBJECTIVE: To observe the correlation between serum level of TXB2, 6-Keto PGF1alpha and liver metastasis. METHODS: The metastatic model was made by injection of W256 carcinosarcoma. Rats were randomly divided into two groups: rats with blood stasis group and control group. Rats in control group were given normal saline via abdominal cavity once a day. Rats in blood stasis group were injected adrenalin in the fourteenth day. Tumor size and liver metastasis were observed. Serum TXB2 and 6-Keto-PGF1alpha were tested by radioimmunoassay. RESULTS: Tumor size in rats with blood stasis was significantly smaller than that of the control group (P<0.01). Occurrence of liver metastasis in rats with blood stasis was significantly lower than that of the control group (P<0.01). The values of 6-Keto-PGF1alpha, TXB2, and TXB2/6-Keto-PGF1alpha were higher in the group with blood stasis. CONCLUSION: In the status of blood stasis, W256 carcinosarcoma grows slowly, and liver metastasis increases insignificantly, with the elevations of 6-Keto-PGF1alpha, TXB2 and TXB2/6-Keto-PGF1alpha. PMID- 15339562 TI - [Pingfei Mixture's two-way adjustment to cell proliferation in mice with tumor]. AB - OBJECTIVE: To study the antineoplastic mechanism of Pingfei Mixture. METHODS: Thirty C57BL mice bearing Lewis pulmonary carcinoma were randomly divided into 3 groups: saline control group, Pingfei Mixture group and cisplatin group. Fifteen days later, tumor tissues and spleens were taken out and made into unicellular suspension. Argyrophil staining was taken to carcinoma cells and cultured T cells. KL-2 style cell image analysis system was used to analyze the rate between AgNORs and nuclear region (I.S). RESULTS: There were dense brownish-black granules in tumor cell nuclear of saline control group. The brownish-black granules of Pingfei Mixture group and cisplatin group were less than those of saline control group. The differences of T cell I.S in these three groups were significant. The I.S of Pingfei Mixture was higher than that of the other groups, and the I.S of the cisplatin group was the lowest. CONCLUSION: Pingfei Mixture can inhibit tumor cell proliferation, although the effect is inferior to cisplatin. Pingfei Mixture can also promote T cell proliferation and its effect was superior to cisplatin. PMID- 15339563 TI - [Effects of Yifei Kangliu Oral Liquid on cell cycle and protein-nucleic acid synthesis of experimental lung cancer]. AB - OBJECTIVE: To explore the effect of the traditional Chinese medicine Yifei Kangliu (YFKL) Oral Liquid on the proliferation, cell cycle and protein-nucleic acid synthesis of murine Lewis lung cancer cell and human lung adenocarcinoma cell line SPC-A-1. METHODS: The inhibiting rates of tumor growth were calculated by weighing the weight of tumor inoculated in vivo, combined by counting cancer cells in vitro. The ratio of the cell cycle and exponents of DNA, RNA, and protein were measured by flow cytometry (FCM). RESULTS: The inhibiting rate of tumor growth in the treated group with YFKL Oral Liquid was 30.38% (P<0.05). The proportion of cells in S phase of the treated groups with YFKL Oral Liquid was lower than that of the control group. In the group with most significant result, 72% of the cells were stagnated in G0/G1 phase. The inhibiting rates of DNA, RNA and protein in murine Lewis lung cancer were 7.4%, 23.73% and 23.31% respectively. In SPC-A-1 cell line, the inhibiting rates were 9.3%, 10.1% and 14.7% respectively, demonstrating amplified effects on lower levels. CONCLUSION: YFKL Oral Liquid significantly inhibited the proliferation of murine Lewis lung cancer cell and human lung adenocarcinoma cell line SPC-A-1 by blocking the cancer cells entering the proliferative phase resulted from its inhibition of DNA. PMID- 15339564 TI - [Original studies on anti-tumor and immunological effect of extracts from Thymus quinquecostatus Celak in mice]. AB - OBJECTIVE: To study the anti-tumor and immunological effect of extracts from Thymus quinquecostatus Celak on mice transplanted S180 tumor cells. METHODS: Different doses of volatile oil and alcohol extracted substances from Thymus quinquecostatus Celak were given to mice bearing S180 tumor for 9 days. Tumor inhibition rates and coefficients of spleen and thymus were determined. RESULTS: Tumor inhibition rates of the groups with alcohol extracts (40 g crude drug.kg( 1).d(-1) and 20 g crude drug.kg(-1).d(-1)) were 51.5% (P<0.01) and 36.4% (P<0.05) respectively, and those of the groups with volatile oil (40 g crude drug.kg( 1).d(-1) and 20 g crude drug.kg(-1).d(-1))were both 39.4% (P<0.05). CONCLUSION: The extracts from Thymus quinquecostatus Celak have anti-tumor activities. The coefficient of spleen in group with alcohol extracts (40 g crude drug.kg(-1).d( 1))was close to normal value, and its coefficient of thymus was between that of the negative control group and the group with cyclophosphamide (0.02 g.kg(-1).d( 1)). The anti-tumor activity of the alcohol extracts was significantly higher than that of the control group and the tumor inhibition rate was depending on drug concentration. Depending on index of immunity,the extracts from Thymus quinquecostatus Celak may have some influences on immunity. PMID- 15339565 TI - [Effect of salvianolate on migration of human vascular endothelial cells]. AB - OBJECTIVE: To study the effect of salvianolate on the migration of endothelial cells induced by monocytes. METHODS: Transwell-boydom system was used to test the migration of endothelial cells induced by monocytes. Enzyme-linked immunosorbent assay (ELISA) and RT-polymerase chain reaction (RT-PCR) method were used to determine the effect of salvianolate on the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) and their mRNA of monocytes. RESULTS: The migration of endothelial cells induced by monocytes was facilitated by salvianolate, compared with the control. After the monocytes were treated by salvianolate for 24 hours, the expression of mRNA of VEGF and bFGF was significantly up-regulated, and the expression of VEGF and bFGF was increased. CONCLUSION: The facilitation of salvianolate on the migration of endothelial cells induced by monocytes was observed. Salvianolate stimulates the expression of VEGF and bFGF and their mRNA of monocytes, and salvianolate may have the role of inducing migration of endothelial cells by working on these two factors. PMID- 15339566 TI - [Clinical observation of cervical spondylopathy treated by acupuncture combined with cupping: a report of 66 cases]. PMID- 15339567 TI - [Rapid identification of Cortex moutan radicis and similar aromatic Chinese herbs by headspace gas chromatography]. AB - OBJECTIVE: To establish the method for rapid identification of the quality of Cortex moutan radicis and similar aromatic Chinese herbs by headspace gas chromatography (HSGC). METHODS: The constituents of headspace gas from Chinese herbs, such as Cortex moutan radicis,Radix cynanchi paniculati and root-bark of Paeonia lactiflora. were analyzed by HSGC. RESULTS: The characteristic finger print headspace chromatograms of these three drugs were different from one another. Cortex moutan radicis stored for 4 years under common conditions contained only traces of paeonol. CONCLUSION: HSGC was simple, quick and accurate in identifying and controlling the quality of aromatic Chinese herbs. PMID- 15339568 TI - [ELISA kit for detection of biliary vesicular protein and its preliminary clinical application]. AB - OBJECTIVE: To establish a rapid and precise detective method of 33.5 kd vesicular protein and to screen an effective treatment of cholelithiasis. METHODS: Specific antibody of the biliary vesicular protein was obtained by immunizing rabbits and enzyme-linked immunosorbent assay (ELISA) kit was developed. The concentrations of 33.5 kd vesicular protein in serum and bile of gallstone patients and control were examined respectively. The effects of Cholagogue Dry Syrup and Eulektrol Capsule on decreasing 33.5 kd vesicular protein were also studied by ELISA kit. RESULTS: One-step ELISA equation was Y=0.035 X (r=0.99). The vesicular protein concentrations in serum and bile of cholesterol gallstone group [(179.8+/-97.9) mg/L and (213.4+/-70.1) mg/L respectively] were significantly (P<0.05) higher than in the pigment stone group and control. Data showed that, with 2-week administration, Cholagogue Dry Syrup significantly decreased both biliary and serum 33.5 kd vesicular protein of cholesterol gallstone patients, while Eulekrol Capsule and control groups didn't have the same results. CONCLUSION: The concentrations of 33.5 kd protein are different in cholesterol gallstone patients and healthy groups which might be related to cholesterol nucleation process. Cholagogue Dry Syrup is of cholagogic and litholytic effect by decreasing biliary lithogenesis. PMID- 15339569 TI - [Study on tumor cells' multidrug resistance and its reversion by Chinese herbs]. AB - Multidrug resistance (MDR) is an important biological behavior of tumor cells in chemotherapy. And it is also one of the major causes of clinical chemotherapy failure. According to the literature at home and abroad, and combining with the results of authors' investigations, this paper mainly discusses the mechanism of tumor cells' MDR and its reversion by Chinese herbs. PMID- 15339570 TI - [Progress of research on apoptosis of breast cancer cells]. AB - This article reviewed information related to the apoptosis of breast cancer cells. Breast cancer is a common malignant tumor in women. Apoptosis is related to the response and resistance to treatment in breast cancer. Here we summarized and integrated the data on apoptosis and its role in the development, prognosis, and treatment of breast cancer. Further study on apoptosis in breast cancer can help find more candidates for predictors and indices of response. Such study will also develop treatment regimens that are tailored to individual tumors and thereby maximize survival. PMID- 15339571 TI - [Application of traditional Chinese medicine in interventional treatment of carcinoma]. AB - Interventional therapy is characterized by mini-invasion, accuracy, obvious curative effect and little side effect. In China, interventional therapy has been applied widely in the treatment of malignant carcinoma in recent 10 years. Traditional Chinese medicine (TCM) also has been adopted in the interventional therapy in recent years. This article reviews the history, status quo and prospect in interventional treatment of carcinoma with traditional Chinese medicine in experimental and clinical domains in recent years. Interventional therapy combined with TCM preparation directly or TCM therapy as a supplementary method of interventional therapy for carcinoma has played an active role in improving clinical curative effect, controlling and reducing toxic side effect and complications. PMID- 15339572 TI - [Prevention and treatment of pulmonary-fibrosis by traditional Chinese medicine]. AB - Pulmonary-fibrosis (PF) is the result of interstitial lung disease which has different causes and it is one of the most intractable diseases in respiratory system. The PF is characterized by progressive difficulty of breath and dying of respiratory failure, seriously threatening the health of people. Immunosuppressive drugs and glucocorticoids have been widely adopted as the main methods in treatment, but the results are not satisfactory and the side effects are obvious. Many specialists have tried traditional Chinese medicine in recent years with satisfactory effect, indicating that the prospect of using traditional Chinese medicine in the treatment of PF is optimistic. This article outlines the researches about using traditional Chinese medicine in the treatment of PF. PMID- 15339574 TI - [Development of the integrated TCM hepatology with western medicine]. AB - The treatment for severe acute respiratory syndrome (SARS) that happened epidemically this spring confirmed again that the integrated traditional Chinese and western medicine had its clinical advantage and vitality, and indicated that this integration could benefit the pathologic understanding and therapeutic strategies for the diseases. Based on the experimental research, western medicine focuses on the histological and structural changes of the diseases. In order to probe the key mechanisms of the disease's occurrence and development, it tries to develop the therapy targeting the mechanism. Materialism is the academic characteristic of western medicine. Based on the differentiation of individual patient and comprehensive analysis of all manifestations, with the academic characteristic of dynamics and dialectics, traditional Chinese medicine (TCM) focuses on the regulation and recovery of the body functions. Theoretically, materialism and dialectics are complementary to each other. Their combination and unity is the key point for the integration of TCM with western medicine in order to improve the clinical effectiveness and TCM development. Therefore, we must adhere to the principles of the evidence-based medicine, strengthen the integrated study on "disease" of western medicine and "syndrome" of TCM, insist on "materialism" and improve the understanding of the mechanism of Chinese herb formula, pay efforts on the clinical study of difficult and common hepatic diseases, in order to develop the integrated traditional and western medicine on liver diseases. PMID- 15339575 TI - [Outline for the theory of diagnosis and treatment based on the correlation between consciousness and organ]. AB - Enlightened by the idea of correlation of mind and body in traditional Chinese medicine and western medicine, integrating his long-term clinical experience, the author brings forward a concise and practical theory for diagnosis and treatment based on the correlation between consciousness and organ in this article. This article includes:(1)introducing the conceptions of consciousness and organ;(2) pointing out the diagnostic and remedial meaning of the four kinds of condition about the correlation of sense and organ;(3) pointing out the diagnostic and remedial meaning of the five kinds of condition about the correlation of desire and organ;(4) how to select herbal and mineral drug to treat the disorders of consciousness and organ;(5) citing some examples to prove the diagnostic and remedial effect of the theory on intractable diseases. PMID- 15339576 TI - [Several viewpoints about the pharmacological research on Chinese herb recipe]. AB - Chinese herb recipe is widely applied and plays a key role in traditional Chinese medicine. However, it involves complicated aspects, and the mechanism researches of Chinese herb recipe had few breakthroughs. The author discussed the questions in the scopes as follows: drug mechanism and chemical components, old typical and current experimental recipes, drug action and its pathological mechanisms. The author believed that the recipe with solid clinical effect should be investigated combining the single herb with its chemical components, and combining the pharmacological mechanism of the recipe with the pathological mechanism of the diseases. More attention should be paid to effect of the research on the improvement of the traditional Chinese medical theory, and efforts should be made to investigate the recipe mechanism relating to molecular signal transduction and the key chemical components. PMID- 15339577 TI - [Multicenter clinical study about the action of Fuzheng Huayu Capsule against liver fibrosis with chronic hepatitis B]. AB - OBJECTIVE: To study the efficacy and safety of Fuzheng Huayu Capsule (FZHY Capsule) against liver fibrosis with chronic hepatitis B. METHODS: Multicentric, randomized, double blinded and paralleled control led trial was conducted on patients (aged between 18 and 65) with liver fibrosis in chronic hepatitis B Indexes observed: (1) hepatic histological changes and HBV markers were observed at 0 and 24th week during the treatment; serological indexes (HA, LN, P-III-P, IV C) were determined and B ultrasound examination of spleen and liver was taken at 0, 12th, 24th week; liver function (during the period of follow-up, liver function and serological indexes for liver fibrosis were evaluated) were observed at 0, 6th, 12th, 18th, 24th week; (2) indexes for safety: blood and urine routine tests, renal function and ECG were examined. RESULTS: (1) Enrollment and demographic data: There was no significant difference between the trial (110 cases) and control group (106 cases) in demographic feature, vital signs, course of illness, history for drug anaphylaxis, history of previous therapy, liver function, serological indexes for liver fibrosis, liver histological examination (99 cases for test group, 96 cases for control group), HBV markers, and renal function, etc. (2) Histological pathological examination: 93 cases of liver histological examination were taken, of these 50 cases for the trial group and 43 cases for control group which turned out to be at S mean value of 2.33 and 2.11 respectively pretreatment according to criteria for liver fibrosis staging. Post treatment, the trial showed a significant decrease with S value of 1.80 compared to that of pretreatment; however, there was no significant improvement in control group before and after the treatment with S mean value of 2.14. There was significant difference in reversing rate (decrease at least 1 stage according to criteria for liver fibrosis staging) between the trial (52%) and control (23.3%) after liver biopsy. The trial had a rather good effect on improving inflammatory activity and was superior to control group with a marked decrease of mean value of inflammatory activity and score of inflammation (P<0.05). (3)Serological indexes for liver fibrosis: There was a significant decrease in HA, LN, P-III-P, IV-C content in test group after 12 and 24 weeks' treatment compared to that of pretreatment; the differences of HA, LN, P-III-P, IV-C between 12, 24 weeks' treatment and pretreatment were significantly greater than control group (P<0.01 or 0.05); the effectual was defined as 2 of 4 indexes lowered more than 30% of the baseline, according to this criteria, the trial was 72.7%, while control group 27.4% (P<0.01). (4)Liver function: Obvious improvement of serum Alb, ALT, AST, GGT was seen in 2 groups; compared with control group, marked improvement of GGT and Alb in the trial (P<0.05); the effective rate of serum ALT in the trial group was 72.7%, while control 59.4%. (5)No changes of significant difference between pre- and post-treatment in routine tests for blood and urine, renal function and ECG, etc. There was also no difference in the stable rate of ALT and serological indexes for liver fibrosis between the trial and control group 12 weeks after withdrawal (P<0.05). CONCLUSION: Fuzheng Huayu Capsule has good effect on alleviating liver fibrosis in chronic hepatitis B without any adverse effect and is superior to Heluo Shugan Capsule. Fuzheng Huayu Capsule is a safe and effective medicine for the treatment of liver fibrosis in chronic hepatitis B. PMID- 15339578 TI - [Clinical features of primary biliary cirrhosis and treatment of it by integrated traditional Chinese and western medicine]. AB - OBJECTIVE: To investigate the clinical features of primary biliary cirrhosis (PBC) and its treatment by integrated traditional Chinese and western medicine. METHODS: 16 PBC patients were observed. Ursodeoxycholic acid (UDCA) was used in the dose of 13 to 15 mg.kg(-1).d(-1), with some traditional Chinese herb prescription. Results (1) The proportion of women to men was 15:1, the mean age was 52.5 years. AMA-M2 was positive in 14 subjects (87.5%). Biliary tract enzymes and ESR were elevated in all subjects. The ratio of hypercholesterolemia (CHOL) and abnormality in IgM was high (62.5%). Fatigue, pruritus, arthralgia, jaundice, splenomegaly were noted in more than half cases. Every patient had one to four complications. (2) ALP, GGT and Glb declined evidently after 3 months' treatment by western medicine associated with traditional Chinese medicine, and declined markedly after 12 months' treatment (P<0.05 respectively). TBIL and CHOL declined gradually during the treatment course. Symptoms and signs were lightened. CONCLUSION: PBC has complicated and especial clinical features. UDCA therapy is effective in PBC, while traditional Chinese medicine has extraordinary effect in treating the symptoms and signs. PMID- 15339579 TI - [Clinical study on treatment of alcoholic liver disease by Qinggan Huoxue Recipe]. AB - OBJECTIVE: To study the effect of Qinggan Huoxue Recipe on alcoholic liver disease(ALD). Methods Effects were observed of Qinggan Huoxue Recipe on the symptoms, physical signs, liver function, blood lipid, markers of liver fibrosis, cytokine and ultrasonogram of ALD patients; the control groups were treated with Xiao Chaihu Decoction or general supportive treatment. RESULTS: The effects of Qinggan Huoxue Recipe were better than that of the other treatments on anorexia, nausea, vomiting, jaundice, ALT, AST and TG; the effects of Qinggan Huoxue Recipe on GGT and VLDL were better than that of the general supportive treatment. Qinggan Huoxue Recipe could decrease the level of cytokines and markers of liver fibrosis, its total effective rate was better than that of Xiao Chaihu Decoction and general supportive treatment. CONCLUSION: Qinggan Huoxue Recipe could prevent and cure ALD markedly, its mechanism might be: resisting lipid peroxidation, stabilizing the membrane of the hepatocytes, rectifying the disturbance of liver lipid metabolism, adjusting the immune function, anti-fibrosis, and promoting the metabolism of ethanol in the liver. PMID- 15339580 TI - [Clinical investigation on characteristics of traditional Chinese medical syndrome of hepatocirrhosis]. AB - OBJECTIVE: To explore the characteristics of traditional Chinese medical syndrome (TCM syndrome) of hepatocirrhosis. METHODS: Clinical information from the four diagnosis methods of traditional Chinese medicine (TCM) and related laboratorial indexes were systematically collected from 223 hepatocirrhosis cases, and the multi-statistical methods including systematic cluster analysis, principal component analysis, stepwise discrimination and variance analysis were made with the software SAS 6.11. RESULTS: Multi-analysis showed that there were 3 categories of syndrome characteristics. Type 1 (134 cases): damp heat, blood stasis, deficiency of liver and spleen Qi; Type 2 (62 cases): deficiency of both Qi and Yin with severe deficiency of Qi, heat with severe dampness, blood stasis; Type 3 (27 cases): deficiency of both Qi and Yin with severe deficiency of Yin, stasis and heat or dampness. Analysis of the changes of the related laboratorial indexes among the three types of syndrome showed that Type 1 mainly manifested asthenia syndrome with sthenia syndrome, and its indexes of AST, ALT, GGT levels were markedly higher than those of Type 2 and Type 3, both of which mainly showed sthenia syndrome with asthenia syndrome, and that Type 3 was in active inflammation, deficiency of both Qi and Yin (deficiency of Yin > deficiency of Qi), and its FN, Alb, FV, FVII, PLT, PCT levels were obviously reduced. CONCLUSION: The multi-statistical methods can reveal the characteristics and regularity of TCM syndrome of hepatocirrhosis, and the 3 categories of syndrome characteristics basically conform to clinical manifestations. The result of TCM syndrome distribution and laboratorial indexes infer that damp heat is the pathological basis of hepatocirrhosis, and the degree of liver function disorder and liver damage may be the pathological basis of deficiency of Yin of both liver and kidney. PMID- 15339581 TI - [Study of an animal model of post-menopausal atherosclerosis in rabbit and observation of the effect of herbal formula of tonifying-kidney on it]. AB - OBJECTIVE: To establish an animal model imitating post-menopausal atherosclerosis in rabbit and observe the effect of the herbal formula of tonifying-kidney on this animal model. METHODS: The New Zealand rabbits were treated with ovariectomy, high cholesterol diet plus immune injury, and both of ovariectomy and high cholesterol diet plus immune injury respectively. The blood lipid level and the intima and media of the aorta were observed. RESULTS: The rabbits treated with ovariectomy and high cholesterol diet plus immune injury changed markedly and had low level of estrogen, high levels of CHOL, TG and dysfunction of the arterial endothelium cells. however, the chinese herbal formula of tonifying kidney could relieve these changes. CONCLUSION: The New Zealand rabbits treated with both of ovariectomy and high cholesterol diet plus immune injury could result in the changes imitating post-menopausal atherosclerosis, and it could be improved by the treatment of herbal formula of tonifying-kidney, so we think that the changes of the animal model are related to the deficiency of the kidney in traditional Chinese medicine. PMID- 15339582 TI - [Hepatocyte apoptosis and the expression of Bcl-2 and Bax in Yin-jaundice rats]. AB - OBJECTIVE: To investigate the effects of Yinchen Shufu Decoction on hepatocyte apoptosis and the expression of its apoptosis-regulating gene Bcl-2 and Bax in Yin-jaundice rats. METHODS: Forty-eight rats were divided randomly into 4 groups: the normal control group, Yin-jaundice model group, Yang-jaundice model group, Yinchen Shufu Decoction treatment group. The TUNEL assay and the immunohistochemistry assay were used to detect the apoptosis of hepatocytes and the expression of Bcl-2 and Bax in hepatocytes respectively. RESULTS: The rate of apoptosis cells in Yin-jaundice model group was higher significantly than that in Yang-jaundice model group and normal control group (P<0.01), the expression of Bcl-2 in Yinchen Shufu Decoction treatment group was higher significantly than that in Yin-jaundice model group (P<0.01 or P<0.05), and the expression of Bax in it was lower significantly than that in Ying-jaundice model group (P<0.01 or P<0.05). CONCLUSION: Yinchen Shufu Decoction can prevent hepatocyte apoptosis perhaps by up-regulating the expression of Bcl-2 and down-regulating the expression of Bax. It is one of the mechanisms of its treatment on Yin-jaundice. PMID- 15339583 TI - [Effect and mechanism of Tangshenning Recipe on micro-albuminuria in rats with early diabetic nephropathy]. AB - OBJECTIVE: To study the modifying effect and mechanism of Tangshenning Recipe on micro-albuminuria in rats with early diabetic nephropathy (DN). METHODS: Male Wistar rats were randomly divided into the normal group (n=8) and model group (n=24). Intraperitoneal injecting of streptozotocin (STZ) plus complete Freund's adjuvant (CFA) was applied once a week for 3 times to induce the DN rats model. Three weeks later, the model group rats were randomly divided into pathologic group (n=8), monopril group (n=8) and Tangshenning Recipe group(n=8) according to the 24 h U-Alb. Each group's renal hemodynamics index and SOD, GSH, MDA in renal tissue were determined by radioimmunoassay (RIA) and colorimetric method respectively. RESULTS: The levels of plasmatic TXB(2), the ratio of TXB(2) and 6 keto-PGF1alpha, and the CGRP in pathologic group were significantly higher than those in normal group. The levels of plasmatic ET decreased obviously, SOD decreased and MDA increased significantly in the rats' renal tissue of pathologic group. The levels of plasmatic TXB(2), the ratio of TXB(2) and 6-keto-PGF1alpha decreased significantly in both Tangshenning Recipe group and monopril group, and the therapeutic effect of Tangshenning Recipe group was better than that of monopril group. SOD was higher and MDA was lower in Tangshenning Recipe group than that in pathologic group. CONCLUSION: The results indicates that Tangshenning Recipe can lower the micro-albuminuria in early DN rats, the mechanism of which probably lies in the modification of glycometabolism, the ratio of TXB(2) and 6-keto-PGF1alpha, the plasmatic CGRP and the renal lipid preoxidation. PMID- 15339584 TI - [Influence of Shenqi Recipe on mesangial cells and interstitial fibroblasts of chronic renal failure rats]. AB - OBJECTIVE: To observe the effect of Shenqi Recipe on the proliferation of glomerular mesangial cells (MC) and interstitial fibroblasts (FC) in rats. METHODS: The effect of serum containing prepared drugs from normal and chronic renal failure rats treated by Shenqi Recipe on glomerular mesangial cells and intersititial fibroblasts in vitro was observed by MTT colorimetric assay and serophrmacology. RESULTS: Compared with the blank sample normal rats, serum containing Shenqi Recipe inhibited the proliferation of MC and FC (P<0.001), and the serum containing Shenqi Recipe of CRF rats had significantly inhibited the proliferation of MC and FC compared with the model group(P<0.001). Conclusion Shenqi Recipe can postpone CRF progress by this mechanism. PMID- 15339585 TI - [Effect of Boschniakia rossica extract on free radicals in brain of D-galactose induced senile rats]. AB - OBJECTIVE: To study the effect of Boschniakia rossica extract on free radicals in the brain of D-galactose induced senile rats. METHODS: Sixty Wistar rats were randomly divided into normal group, model group (48 mg.kg(-1).d(-1) D-galactose, SC), Boschniakia rossica group (100, 150, 200 mg/kg ig and 48 mg.kg(-1).d(-1) D galactose, SC). After 40 days, the activities of SOD, MAO and the content of MDA were measured with colorimetric method, and the histological changes were synchronously observed by electronic microscope. RESULTS: Boschniakia rossica extract significantly increased the SOD activity, decreased the MDA content, and inhibited the MAO activity in the brain tissue. It was observed under microscope that Boschniakia rossica extract could retrieve the degeneration of mitochondrion. CONCLUSION: Boschniakia rossica extract can clear the free radicals for D-galactose induced senile rats. PMID- 15339586 TI - [Inhibition of tumor growth in vitro by the extract of Fagopyrum cymosum]. AB - OBJECTIVE: Fagopyrum cymosum (Trev.) Meisn has long been used in China to treat various ailments of the lung, including lung tumors. This study investigated whether Fagopyrum cymosum extract (Fago-c) has effects on other organs. METHODS: Human cancer cells derived from 10 different organs were employed, and their growths as affected by Fago-c were investigated. RESULTS: It was found that the growth of cancer cells from lung, liver, colon, leukocytes and bone is inhibited by Fago-c. However, cancer cells derived from prostate, cervix, ovary and brain are not sensitive to Fago-c, and the extract stimulates the growth of cancer cells from breast (MCF-7). Synergistic inhibition effect of Fago-c and daunomycin was observed in human lung cancer cells (H460). Cellular proteins from H460 cells treated with Fago-c were analyzed by 2D-gel electrophoresis. A protein (M.W./pI=20 K/5.9) was induced. The Fago-c extract was analyzed by High Performance Liquid Chromatography (HPLC). Four major and twenty minor components were identified. CONCLUSION: These studies indicate the effect of Fago-c in inhibiting the growth of cell lines derived from certain organs. PMID- 15339587 TI - [Study on estrogen receptor of adult human osteoblasts]. AB - OBJECTIVE: To examine estrogen receptor (ER) in osteoblasts from adult human and to elucidate the mechanism of estrogen in modulating bone metabolism. METHODS: The cultured osteoblasts were harvested from bone chips by modified sequential digestive enzyme release and immunohistochemical assay of ER in osteoblasts were carried out in three groups of female adults: normal control (group 1), patients with moderate osteoporosis (group 2) and patients with serious osteoporosis (group 3). The percentages of ER-positive osteoblasts from the three groups were compared by t test. RESULTS: The brown marks that indicate ER were found in nuclei and plasma of the osteoblasts, and the percentages of ER-positive osteoblasts among three groups were significantly different. CONCLUSION: ERs exist in nuclei and plasma of the osteoblasts. Estrogen may modulate bone metabolism through binding ER in nuclei and plasma of the osteoblasts. The reduction of ER of osteoblasts may play an important role in the pathogenesis of postmenopausal osteoporosis. PMID- 15339588 TI - [Study on the application of bio-heat transfer science to the quantitative analysis of tongue inspection]. AB - OBJECTIVE: To research the mechanism of the tongue inspection in traditional Chinese medicine and to study the special relationship between temperature distribution of the tongue and the tongue color. METHODS: The temperature field of human and pig's tongue was measured by infrared thermal vision system and thermal-couple probe. The blood perfusion rates in the human tongue and the pig's tongue were obtained with laser Doppler rheometer and CBI-8000 physiology research system. The temperature field of the profile of tongue was computed by numerical method. RESULTS: The quantitative curve of the temperature distribution on the surface of tongue and the relationship between temperature and blood perfusion rate were obtained. And the temperature in the profile of tongue was obtained by finite element method. CONCLUSION: The research involves that the different tongue color can reflect different tongue temperature and blood perfusion rate. This proved that it is acceptable to apply the bio-heat transfer theory to the tongue inspection research. The results provided a new method to research the tongue inspection in traditional Chinese medicine. PMID- 15339589 TI - [Studies on treatment of fatty liver with traditional Chinese medicine]. AB - Great progress has been made in the study of fatty liver with integrated traditional Chinese and western medicine in aspects of diagnosis, treatment and experimental study, etc. Most researches were designed to utilize diagnostic or model replicating method of western medicine to observe the effects or investigate the action mechanism of compound recipe, single Chinese herb or effective ingredients of Chinese herbs on fatty liver. According to the pathological mechanism of traditional Chinese medicine (TCM), fatty liver is characterized by deficiency in nature and repletion in appearance, which involves three Zang viscera such as liver, spleen and kidney and manifests as spleen Qi deficiency, liver and kidney deficiency, phlegm and dampness heaping internally, and Qi stagnation and blood stasis. This facilitates us to use specific recipe or modified recipe to treat fatty liver from the points of integrated traditional Chinese and western medicine and combining syndrome differentiation with disease differentiation. With gratifying achievement, this kind of approach has been the mainstream of the research on fatty liver and many researchers have reached an agreement on this point domestically. Spleen Fortifying and Blood Invigorating Recipe (SFBVER in brief, invented by our institute) can significantly improve the B ultrasound outcome of the liver in patients with fatty liver, with significant difference in B ultrasound scoring between pre-and post-treatment. It can alleviate the patients' symptoms, improve or regain liver function, decrease waist/buttocks ratio and the content of triglyceride and cholesterol in blood. SFBVER is superior to Dongbao Gantai Recipe in general effective rate. Experimental study also reveals that SFBVER can alleviate CCl(4) induced liver cell fatty degeneration and the inflammatory cell infiltration in rats, decrease the activities of ALT and AST, lower the content of triglyceride in liver, recover SOD activity in liver to normal level. The overall efficacy of SFBVER is superior to that of Dongbao Gantai Recipe. Further correlated study should be focused on inventing new preparation of traditional Chinese medicine and investigating its action mechanism with the guiding of the theory of TCM and referring to the latest discovery in fatty liver research in modern medicine. PMID- 15339590 TI - [Chronic gastritis treated by acupuncture combined with Chinese herbs: a report of 110 cases]. PMID- 15339591 TI - [Advances of mechanism research on treatment of experimental liver fibrosis with traditional Chinese medicine]. AB - The paper reviewed and analyzed the literature in the past five years about the function of the cells and mediums in the liver, and the influence and mechanism of traditional Chinese drug therapy on experimental liver fibrosis. It reveals that the traditional Chinese drug therapy treats experimental liver fibrosis through lots of mechanisms. The author gives also some suggestions on the further research into liver fibrosis. PMID- 15339592 TI - [Advances of combined treatment for chronic hepatitis B with lamivudine]. AB - This paper summarized the effects of the treatment of chronic hepatitis B (CHB) by lamivudine combined with traditional Chinese medicine or western medicine in the past few years. Combined treatment of lamivudine and the other medicine had higher efficiency than lamvudine alone in the treatment of CHB. The Combined treatment should be a tendency of the treatment for chronic hepatitis B. The focal point is that the traditional Chinese recipe produces a multitarget effect in the treatment of CHB. The combined treatment of lamivudine and traditional Chinese recipe can reduce the treatment course and the recurrence rate. Combined treatment of traditional Chinese medicine and western medicine has notable superiority and broad developing prospect in the treatment of CHB. PMID- 15339593 TI - [Congenital meningomyelocele complicated by incontinence of urine following the burn of the four limbs: a report of one case]. PMID- 15339594 TI - [Evidence-based medicine and treatment of lung cancer]. AB - Traditional Chinese medicine (TCM) thinks highly of Zheng Hou in the treatment of lung cancer. Zheng is evidence. Hou is clinical manifestations. After obtaining information and evidence of patients by means of the four diagnostic methods, doctors treat patients by syndrome differentiation. In TCM, the collection and study of medical literature of tumor are emphasized. Medical classics and successful treatment of certain cases are used to guide clinical treatment. Such practice of TCM is similar to that of evidence-based medicine (EBM). Oncology of TCM and EBM are all of ancient origin. There are many clinical reports of satisfactory curative effects in treating lung cancer by TCM. But there are still many problems in methodology, such as careless random method, incomparability in base line, and deficiency of large exponent, multicenter, blind method or randomization. Sometimes the effects of control drugs are incredible. The determination of near effect is not strict. There are exponent errors in statistics of near effect(CR+PR). Certainly, treatment of lung cancer by TCM has its uniqueness. The main characteristic of treatment by TCM is that it can restrain or slow the growth of tumor. At the same time, physical signs, symptoms and quality of life can be improved, and survival period is prolonged. "Survival rate is the golden standard". If we can explore a new treatment strategy to control the growth and metastasis of advanced NSCLC and prevent relapse and metastasis of earlier period or metaphase NSCLC by the method of binding TCM and west medicine under the rules of EBM, such as multicenter (TCM hospital and western medicine hospital), blind method, strict randomization and control, the conclusion of the study will be more scientific. PMID- 15339595 TI - [Comments on treatment of severe acute respiratory syndrome by integrated traditional Chinese and western medicine]. AB - There are obvious advantages of the integrated traditional Chinese and western medicine on the treatment of severe acute respiratory syndrome (SARS). Combining the ancient and present literature with the special epidemic patterns, pathological changes and clinical symptoms of SARS, the paper discussed the methods of application of the integrated traditional Chinese and western medicine on the treatment of SARS, and some matters needing attention in clinic. PMID- 15339596 TI - [Zhang Yun-Peng's treatment of one case of SARS based on syndrome differentiation]. PMID- 15339597 TI - [Abortion induced by drug with the addition of Shenghua Decoction: an analysis of 100 cases]. PMID- 15339598 TI - [Superiority of traditional Chinese medicine on gene regulation]. PMID- 15339599 TI - [Treatment of uveitis with integrated traditional Chinese and western medicine: a report of two cases]. PMID- 15339600 TI - [Towards integrative postmodern individualized medicine in terms of hierarchic emergence]. AB - The paper discusses the possibility and necessity for establishing a postmodern individualized medicine in terms of hierarchic emergence, a basic concept of contemporary system science. Through analysis of the strong and weak points of modern individualized medicine, personalized medicine and genomic medicine, the author found that Chinese traditional individualized medicine has its strong points in implementing whole (holistic) emergence and weak points in lower hierarchic emergence compared with the other three modern medical disciplines. Thus the author concluded that there are possibility and necessity to integrate the strong points of these medical systems for establishing a holistic and deepgoing patient-centered postmodern individualized medicine. PMID- 15339601 TI - [Curative effect of cyclic adenosine monophosphate and Shengmai injection on the aged patients with sinus bradycardia]. PMID- 15339602 TI - [Puzzle of hormone replacement therapy and prospect of the role of traditional Chinese medicine in treating postmenopausal syndrome]. AB - In July 2002, the Women's Heath Initiative (WHI) clinical trial, designed to clarify the risks and benefits of combination hormone replacement therapy (HRT) to the postmenopausal women declared that interim safety review after an average follow-up of 5.2 years found that a combination of estrogen and progestin frequently prescribed to postmenopausal women in USA increased the risk of invasive breast cancer, heart disease, stroke, and pulmonary embolism while reduced bone fractures and colorectal cancer. The overall risks of HRT outweigh the benefits, which provides an opportunity for traditional Chinese medicine (TCM) going abroad. A variety of clinical and experimental evidences have showed that TCM exerts quite satisfactory effect on relieving postmenopausal symptoms with little adverse effect, hence a potential role to replace or to improve HRT or to reduce the side effect induced by HRT. PMID- 15339603 TI - [Experience in treating biliary tract diseases]. PMID- 15339604 TI - [Supplementing Qi and activating blood circulation in gene regulation of vascular smooth muscle cells]. AB - According to the theory of traditional Chinese medicine, blood stasis is a main pathological mechanism in the development of vascular diseases. Supplementing Qi and activating blood circulation, as the therapeutic principle for the blood stasis, has been adapted. Studies demonstrated that the supplementing Qi and activating blood circulation recipe could regulate the expression of vasoactive peptides in vascular smooth muscle cells (VSMCs). The recipe inhibited the formation of neointima following arterial intimal lesions through down-regulating expression of proliferation-related genes and phenotypic modulation of VSMCs. The recipe also markedly inhibited the adhesion and migration of VSMCs and matrix remodelling by means of a mechanism that balances extracellular matrix turnover rate. The purpose of this review is to summarize the recent advances made in our understanding of new functions for the recipe in regulating VSMCs behaviours and their microenvironment relevant to vascular diseases and maintaining proper homeostasis. PMID- 15339605 TI - [Monomer drug's special effect and application of western medicine based on the theory of traditional Chinese medicine]. AB - The concepts of monomer medicine and the Chinese traditional medicine were introduced in this article. The different effects of a monomer drug between different types of syndromes of traditional Chinese medicine (TCM) were explained based on the author's clinical experience and related literature. The aim, significance and methods about the application of western medical drugs according to the theory of TCM were pointed out. And the feasibility of the application was analyzed. PMID- 15339606 TI - [Study of genes related with kidney deficiency syndrome]. AB - Study of kidney deficiency syndrome may utilize the modern molecular biology technique, such as gene chip, representational difference analysis (RDA) and genes sequence analysis, to clone the different genes of kidney deficiency syndrome from single TCM syndrome type of the disease, to investigate the inner connection between the various phenotypes of kidney deficiency syndrome and their genes expression pattern and their function products, then to definite the related genes of kidney deficiency syndrome. The research method may also be used for the study of other TCM syndrome types. PMID- 15339607 TI - [Research on quantified diagnosis and combining diseases with syndrome of blood stasis]. AB - OBJECTIVE: To explore the way of quantified diagnosis of blood stasis syndrome (BSS) and the essence of BSS. METHODS: Using t-test and logistic regression to analyze the symptoms, signs, and objective indexes of BSS in clinical test. RESULTS: (1) The levels of endothelin, nitric oxide, and t-PA between 182 patients with BSS and non-BSS had notable difference. (2) The stepwise regression analysis about hemoglobin, triglyceride, total cholesterol, endothelin, nitric oxide, and t-PA in 170 patients with BSS or non-BSS revealed that the order of these indexes considering their importance was: endothelin, hemoglobin, t-PA, nitric oxide. However, the triglyceride and total cholesterol could not enter the regression equations. (3) The stepwise regression analysis about 40 symptoms and signs in 601 patients with BSS or non-BSS revealed that the 18 items had the very contribution to diagnose the BSS when the F value was 6, and so a regression equation was available. The total coincidence ratio was 94.24% when the other quantified data about BSS were tested in the equation. CONCLUSION: The standardized TCM syndrome, which accorded with the criteria of combining diseases with syndrome, was made up of symptoms, signs, and objective indexes. The standard could be obtained by multi-center, prospective, random and controlled clinical epidemiological survey and clinical test. PMID- 15339608 TI - [Clinical observation of the curative effect of Qiangxin Mixture on congestive heart failure]. AB - OBJECTIVE: To investigate the efficacy of Qiangxin Mixture in patients with congestive heart failure (CHF). METHODS: Sixty cases of CHF were randomly divided into Qiangxin Mixture group (treatment group, n=30) and digoxin group (control group, n=30). The total clinical effective rate, integra of the symptoms of kidney deficiency, classification of functional capacity of the New York Heart Association (NYHA), and echocardiogram [ejection fraction (EF), cardiac output (CO), interventricular septal thickness (IVST), posterior wall thickness (PWT), left ventricular mass index (LVMI)] as well as the measurement of plasma endothelin, angiotensin II and atrial natriuretic peptide were observed in both groups. RESULTS: The total clinical effective rate of Qiangxin Mixture group was 87%, and improvement was significantly observed in the Lee CHF score, classification of functional capacity of the NYHA, EF and CO (P<0.05, vs before treatment), but no significant improvement in digoxin group (P>0.05). The integra of the symptoms of kidney deficiency, the levels of plasma ET, Ang II and ANP decreased significantly (P<0.01, vs before treatment and digoxin group respectively). IVST, PWT and LVMI were also reduced significantly (P<0.01, vs before treatment; P<0.05 vs digoxin group). CONCLUSION: The Qiangxin Mixture is effective in enhancing cardiac contraction, improving hemodynamics in the short term and rectifying some indexes in the long-term, so it could postpone the processes of CHF. This mechanism may be related to decreasing the stimulating factors (angiotensin, endothelin) which trigger the cardiac remodeling, delaying or reversing the cardiac remodeling. PMID- 15339609 TI - [Evaluation of therapeutic effects of traditional Chinese medicine combined with western medicine on breast cancer after resection: analysis of 71 cases]. AB - OBJECTIVE: To evaluate the effectiveness of traditional Chinese medicine (TCM) combined with western medicine on breast cancer after surgical resection. METHODS: Seventy-one patients with breast cancer received chemotherapy, radiotherapy, endocrine therapy and TCM following resection. RESULTS: None of the patients died during the follow-up period. Fifty-seven patients were treated with TCM shortly after undergoing resection. Two recurrences (3.5%) and 6 metastases (10.5%) were observed in 8 patients, whose median disease free survival was 5.9 years. Fourteen patients didn't receive TCM until recurrence or metastasis emerged, with a median disease free survival of 3.5 years. There was significant difference between the two groups (P=0.033). CONCLUSION: TCM combined with western medicine is efficient in preventing and delaying recurrence and metastasis. PMID- 15339610 TI - [Clinical study and electron microscope observation of tapeworm scolices on 548 cases of taeniasis treated with Binlang Chengqi Decoction]. AB - OBJECTIVE: To observe the effect and mechanism of Chinese herbs in the treatment of taeniasis. METHODS: Five hundred and forty-eight cases of taeniasis were treated with Binlang Chengqi Decoction (BLCQD). The tapeworm scolices of ten cases were observed by electron microscope (EM). RESULTS: Among the 548 cases, 521 cases were cured and 27 cases were improved. The total effective rate was 100%. Foam-like secretion in the sucker of the tapeworm scolices and erosion of the epithelium in the cervical part were observed by scanning electron microscope. Observed by transmission electron microscope, the cortex was seriously damaged. The microvilli were exfoliated. The cells in the muscle layer and parenchyma layer were injured in various degrees. The mitochondria were tumefied or caved. And the nerve cord was damaged. CONCLUSION: BLCQD can not only paralyze the tapeworm scolex, but also injury the cells of the tapeworm scolex. PMID- 15339611 TI - [Operated diabetes insipidus treated by supplementing Qi and activating blood circulation and warming Yang to reinforce]. PMID- 15339612 TI - [Gene expression related to growth axis and gonadal axis in puberty rats]. AB - OBJECTIVE: To elucidate the effect and effective mechanism of gonadotropin releasing hormone analog (GnRHa) on growth axis and gonadal axis in puberty rats. METHODS: After injecting GnRHa on puberty rats, hypothalamus, pituitary, ovary and epiphyseal cartilage of legs were taken, and related gene expression was detected by fluorescence quantitative PCR (FQ-PCR). RESULTS: GnRHa could reduce the gene expression of gonadotropin-releasing hormone (GnRH) in hypothalamus and its receptor (GnRHR) in pituitary, increase the gene expression of growth hormone releasing inhibiting hormone (SRIH), and not affect the gene expression of growth hormone releasing hormone (GHRH) in hypothalamus. And GnRHa could reduce the gene expression of growth hormone (GH) in pituitary, estrogen receptor (ER) in ovary and insulin-like growth factors-I (IGF-1) in epiphyseal cartilage of legs. CONCLUSION: Besides restraining the gene expression of GnRHR in pituitary and leading to descent regulation of receptor, GnRHa can reduce the gene expression of GnRH in hypothalamus and down-regulate gonadal hormone level. At last, GnRHa can reduce the maturity degree and pace of the second sexual characteristics. By means of increasing the gene expression of SRIH in hypothalamus, GnRHa can reduce the gene expression of GH in pituitary and IGF-1 in epiphyseal cartilage of legs. It could be one of the mechanisms of GnRHa reducing epiphyseal maturity for the patients with idiopathic central precocious puberty. PMID- 15339613 TI - [Study of the mechanism of preventing proteinuria by Yishen Gujing Decoction (YSGJD) in early diabetic nephropathy]. AB - OBJECTIVE: To study the mechanism of preventing proteinuria by traditional Chinese medicine YSGJD in early diabetic nephropathy (DN) in rats. METHODS: The male Sprague-Dawley (SD) rats were randomly divided into three groups: normal control, untreated control and treated groups. Experimental diabetes mellitus (DM) was induced in untreated control and treated rats with streptozotocin (STZ), and the normal group was given equal buffer solution. The treated rats were given YSGJD daily. The other two groups were given running water. Eight weeks later, the rats were raised in metabolic cages for 24 hours. Then, the 24 h urinary protein, serum glucose and nitric oxide (NO) were measured. In the renal cortex, the levels of iNOS mRNA and cGMP were determined by reverse transcription polymerase chain reaction (RT-PCR) and radioimmunoassay separately. RESULTS: Compared with normal control, in DN rats, the 24 h urine protein, the levels of renal cortex iNOS mRNA and cGMP were increased significantly (P<0.01). All above in treated group were reduced than in untreated control (P<0.05). The serum glucose and NO were increased in DN rats than in normal control (P<0.01), and had a little decrease in the treated group, with no significance between untreated and treated groups. CONCLUSION: This recipe can diminish the urine protein in early DN without the significant decrease of serum sugar, can correct the abnormal levels of iNOS mRNA and cGMP in renal cortex. Interfering with the function of NO may be one of its mechanisms. PMID- 15339614 TI - [Study on Realgar inducing apoptosis in T lymphocytic cell line CEM]. AB - OBJECTIVE: To study the Realgar induced T lymphocytic leukemia cell line CEM apoptosis in vitro. METHODS: CEM cells were incubated with Realgar. Cell proliferation inhibition was determined by MTT. Cell cycle, apoptosis, Apo2.7 and Fas were measured by cytometer. RESULTS: Realgar inhibited the proliferation of CEM cell line. The cells treated with Realgar showed a Sub-G(0)/G(1) apoptotic peak in DNA distribution histogram, increment of Apo2.7 protein expression, and arrested cells in G(2)/M phase, but ineffectiveness on Fas expression. CONCLUSION: The Realgar can induce CEM cell apoptosis in vitro. PMID- 15339615 TI - [Effect of Sophoricoside on histomorphology of bone in ovariectomized rats]. AB - OBJECTIVE: To evaluate the potential preventive effect of Sophoricoside on bone loss in ovariectomized rats. METHODS: Female SD rats (n=50, 6 months old) were either sham-operated (SHAM group, n=10) or ovariectomized (n=40). Three days after operation, ovariectomized rats were randomly assigned to groups as follows: 10 received Sophoricoside 4 mg.kg(-1).d(-1) (SL group), 10 received Sophoricoside 8 mg.kg(-1).d(-1) (SM group), 10 received Sophoricoside 16 mg.kg(-1).d(-1) (SH group) and 10 were untreated (OVX group). After Sophoricoside were given orally for one month, the histomorphometric parameters in the secondary spongiosa of proximal tibia and lumbar vertebrae were examined. RESULTS: Compared with OVX group, SM and SH two kinds of treatment caused 15.28% and 22.81% increment in percent trabecular area (BV/TV, %) in the secondary spongiosa of proximal tibia, 14.23% and 21.2% increment in lumbar vertebrae. Accordingly in these two groups, there was a significant decrease in trabecular separation (FLAW, microm), but increment in trabecular width (Tb. Th, microm) and conjunction points (JOINT). But in SL group, the preventive effect was not observed. CONCLUSION: Sophoricoside can be efficient in preventing ovariectomy-induced bone loss in rats. PMID- 15339616 TI - [Effect of Sorbaria Sorbifolia extract on anti-oxidative activities in rats with precancerosis induced by diethylnitrosamine]. AB - OBJECTIVE: To study the effect of Sorbaria Sorbifolia extract on anti-oxidative activities in rats with precancerosis induced by diethylnitrosamine. METHODS: Sorbaria sorbifolia was perfused into stomach of rats with DEN-induced precancerosis, then the biochemical indexes were investigated. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and the concentrations of malondialdehyde (MDA), glutathione (GSH) were investigated by colorimetric method. RESULTS: Sorbaria sorbifolia extract increased SOD, GSH-PX and decreased MDA, GSH levels in serum and liver homogenate. CONCLUSION: Sorbaria Sorbifolia extract significantly increased anti-oxidative activities in rats with precancerosis induced by diethylnitrosamine. PMID- 15339617 TI - [Treatment of nodular vasculitis based on syndrome differentiation: a report of 32 cases]. PMID- 15339618 TI - [Comparative study on immune enhancement effects of four kinds of dodder seeds in Shandong Province]. AB - OBJECTIVE: To compare the effects of immune enhancement, anti-fatigue and anoxia tolerance on mice of four kinds of dodder seeds in Shandong Province. METHODS: Water extracts and alcohol extracts of the four kinds of dodder seeds were administered to mice. The phagocytosis of coelio-macrophage, the weight changes of the immune organs, the survival time to swimming and lacking of oxygen were observed. RESULTS: The four kinds of dodder seeds in Shandong Province could enhance the phagocytosis of macrophage of mice and increase the weights of thymus and spleen of the immature mice. They could prolong the survival time of mice swimming and lacking oxygen. Cuscuta chinensis Lam. and C. australis R. Br. had better effects than the other two kinds, and the water extracts had better effects than alcohol extracts. CONCLUSION: The four kinds of dodder seeds in Shandong Province have the effects of immune enhancement, anti-fatigue and anoxia tolerance. The polysaccharide in dodder seeds is one of the effective materials to improve the immune system. PMID- 15339619 TI - [Treatment of acute gastrectasia following liver transplantation with the therapeutic methods of integrated traditional Chinese and western medicine: a report of one case]. PMID- 15339620 TI - [Comparative experimental study on treatment of rat's injured sciatic nerve with electroacupuncture and Buyang Huanwu Decoction]. AB - OBJECTIVE: To study the effect of the electroacupuncture and Buyang Huanwu Decoction (BYHWD) on the regeneration of the rat's injured sciatic nerve. METHODS: Rat's sciatic nerve injury model was made by operation. The rats were treated with electroacupuncture or BYHWD and their nerve electro-physiological indices and histological changes tested by HRP were observed. RESULTS: It was demonstrated that the recovery ratios of nerve conduction velocity and muscle evoked action potential of the group treated with electroacupuncture and the group treated with BYHWD were higher than those of the group treated with western medicine or the group without treatment. The differences were significant. The labeling cells in anterior horn of spinal marrow and spinal ganglion of the group treated with electroacupuncture and the group treated with BYHWD were more abundant than those of the group treated with western medicine or the group without treatment. The differences were significant. The effect of electroacupuncture was better than that of the BYHWD. CONCLUSION: It suggests that both electroacupuncture therapy and BYHWD can promote the recovery of the function in early stage of injured nerve, and they are effective methods for promoting the regeneration of injured peripheral nerve. PMID- 15339621 TI - [Culture of human osteoclasts]. AB - OBJECTIVE: To establish a stable, useful culture system for human osteoclasts and to investigate the effect of osteoblasts on the differentiation, proliferation and activation of osteoclasts so as to provide a base for the studies on prevention and treatment of osteolysis and osteoporosis. METHODS: In the presence of 1,25-(OH)2D3, monocytes abstracted from human bone marrow were cultured in three groups: co-culture of monocytes and osteoblasts, monocytes alone, monocytes with conditional media (CM) of osteoblasts. Differentiation process of the cultured cells was observed under biological microscope. HE staining and tartrate resistant acid phosphatase (Trap) staining were employed to assay the cultured cells. The resorption pits on bone slices, on which cells were cultured, were observed under scanning electronic microscope (SEM). RESULTS: In the group of co culture of monocytes and osteoblasts, monocytes gradually fused to form multinucleated cells (MNCs), and the MNCs were also indicated in HE staining and Trap staining. The SEM showed a number of resorption pits on bone slices. In the other two groups, Trap-positive MNCs were not obtained, and resorption pits were not observed on bone slices. CONCLUSION: In this culture, monocytes obtained from human marrow fused to form multinucleated cells (MNCs) that express the main characteristics of the osteoclast phenotype, such as Trap-positive and the ability to form resorption lacunae when cultured on bone slices. Cell-to-cell contact with osteoblasts was necessary for the differentiation, proliferation and activation of osteoclasts. PMID- 15339622 TI - [Prolapse of lumbar intervertebral disc treated by needling the side of abdomen and fumigation with Chinese herbs]. PMID- 15339623 TI - [Morphological changes following airway remodeling in asthmatic rats]. AB - OBJECTIVE: To explore the establishment of an asthmatic model with airway remodeling in rats by observing the morphological changes of the airway in different stages. METHODS: Animals were divided into two groups: asthmatic group and control group. The subjects were observed at 5 phases: before activation and 2, 4, 6, 8 weeks after activation. General histological changes were observed using hematoxylin and eosin stained sections. Being standardized by internal perimeter (Pi), the wall thickness (d), internal area (Ai), outer area (Ao) and wall area (WA) of the airway were quantified by computer-assisted image analysis system. And the membrane airways were divided into 3 grades (large, media and small airways) to be analyzed. RESULTS: The changes of small airway appeared earlier and greater than the others. The changes of media airway appeared later than the small one. The changes of large airway appeared as later as the 4th week after activation. There were no differences before and after activation in control group (P>0.05). CONCLUSION: (1) Prolonged, repeated and low density Ag activation could establish asthmatic model with airway remodeling. (2) The morphological changes of the airways increased following airway remodeling, and the greatest increase occurred at small airway. (3)The morphological changes in the model rats were similar to that in human beings. PMID- 15339624 TI - [Experience of ZHANG Jian-Qiu in treating "extraordinary meridian diseases" based on syndrome differentiation of miscellaneous diseases]. PMID- 15339625 TI - [Researches into the modernization of tongue diagnosis: in retrospect and prospect]. AB - With the further development of clinical modernization of traditional Chinese medicine (TCM), new request to tongue diagnosis for objectivity and quantification is raised. Many researchers have devoted themselves to this field and have achieved some accomplishments since 1950's. Especially in recent years, the revolution of information technique derived from computer industry brings the research new chances. The research history of the past five decades can be divided into three stages: (1) looking for micro-markers by newly-developed medical technique; (2) quantifying traditional markers by new technique; (3) further development based on information technique and corporation of multiple subjects. This article reviewed the three stages and described its prospect. PMID- 15339626 TI - [The study of intoxication and toxicity of Fructus Xanthii]. AB - Fructus Xanthii is commonly used in rhinology, especially used in treatment of chronic rhinitis and sinusitis. But Fructus Xanthii is toxicant, and it maybe result in poisoning when used excessively, without preparation or in inappropriate preparation. This article reviewed all poisoning cases from 1960 to 2000, its toxicology and methods for detoxification PMID- 15339627 TI - [Pathogenetic characteristics and therapeutic approaches of aplastic anemia]. AB - According to TCM theory of the wholistic concept and the principles of syndrome differentiation and treatment, the characteristics of the pathogenesis of aplastic anemia (AA) were summarized and corresponding treatment was put forward on the basis of clinical observation and the study of literature. In the pathogenesis of AA, the deficiency of spleen and kidney is the primary aspect, fire attacking blood vessels is the secondary aspect and there is change because of blood stasis. In treatment, the method of invigorating spleen and tonifying kidney is adopted in order to get at the root, and releasing fire to stop bleeding is adopted in order to bring about a temporary solution, and activating blood circulation to dissipate blood stasis is adopted in order to produce blood. The outline should be caught hold of that consumption is primary aspect and fire is secondary aspect and there is change because of blood stasis in order to obtain satisfactory curative effect. PMID- 15339628 TI - [Observation of the curative effect of Xinjia Yinchenhao Decoction in treating severe chronic hepatitis]. PMID- 15339629 TI - Effects of data structure on the estimation of covariance functions to describe genotype by environment interactions in a reaction norm model. AB - Covariance functions have been proposed to predict breeding values and genetic (co)variances as a function of phenotypic within herd-year averages (environmental parameters) to include genotype by environment interaction. The objective of this paper was to investigate the influence of definition of environmental parameters and non-random use of sires on expected breeding values and estimated genetic variances across environments. Breeding values were simulated as a linear function of simulated herd effects. The definition of environmental parameters hardly influenced the results. In situations with random use of sires, estimated genetic correlations between the trait expressed in different environments were 0.93, 0.93 and 0.97 while simulated at 0.89 and estimated genetic variances deviated up to 30% from the simulated values. Non random use of sires, poor genetic connectedness and small herd size had a large impact on the estimated covariance functions, expected breeding values and calculated environmental parameters. Estimated genetic correlations between a trait expressed in different environments were biased upwards and breeding values were more biased when genetic connectedness became poorer and herd composition more diverse. The best possible solution at this stage is to use environmental parameters combining large numbers of animals per herd, while losing some information on genotype by environment interaction in the data. PMID- 15339630 TI - Reduction of inbreeding in commercial females by rotational mating with several sire lines. AB - A mating system to reduce the inbreeding of commercial females in the lower level was examined theoretically, assuming a hierarchical breed structure, in which favorable genes are accumulated in the upper level by artificial selection and the achieved genetic progress is transferred to the lower level through migration of males. The mating system examined was rotational mating with several closed sire lines in the upper level. Using the group coancestry theory, we derived recurrence equations for the inbreeding coefficient of the commercial females. The asymptotic inbreeding coefficient was also derived. Numerical computations showed that the critical factor for determining the inbreeding is the number of sire lines, and that the size of each sire line has a marginal effect. If four or five sire lines were available, rotational mating was found to be quite an effective system to reduce the short- and long-term inbreeding of the commercial females, irrespective of the effective size of each sire line. Oscillation of the inbreeding coefficient under rotational mating with initially related sire lines could be minimized by avoiding the consecutive use of highly related lines. Extensions and perspectives of the system are discussed in relation to practical application. PMID- 15339631 TI - A genetic and spatial Bayesian analysis of mastitis resistance. AB - A nationwide health card recording system for dairy cattle was introduced in Norway in 1975 (the Norwegian Cattle Health Services). The data base holds information on mastitis occurrences on an individual cow basis. A reduction in mastitis frequency across the population is desired, and for this purpose risk factors are investigated. In this paper a Bayesian proportional hazards model is used for modelling the time to first veterinary treatment of clinical mastitis, including both genetic and environmental covariates. Sire effects were modelled as shared random components, and veterinary district was included as an environmental effect with prior spatial smoothing. A non-informative smoothing prior was assumed for the baseline hazard, and Markov chain Monte Carlo methods (MCMC) were used for inference. We propose a new measure of quality for sires, in terms of their posterior probability of being among the, say 10% best sires. The probability is an easily interpretable measure that can be directly used to rank sires. Estimating these complex probabilities is straightforward in an MCMC setting. The results indicate considerable differences between sires with regards to their daughters disease resistance. A regional effect was also discovered with the lowest risk of disease in the south-eastern parts of Norway. PMID- 15339632 TI - Genetic parameters for litter size in sheep: natural versus hormone-induced oestrus. AB - The litter size in Suffolk and Texel-sheep was analysed using REML and Bayesian methods. Litters born after hormonal induced oestrus and after natural oestrus were treated as different traits in order to estimate the genetic correlation between the traits. Explanatory variables were the age of the ewe at lambing, period of lambing, a year*flock-effect, a permanent environmental effect associated with the ewe, and the additive genetic effect. The heritability estimates for litter size ranged from 0.06 to 0.13 using REML in bi-variate linear models. Transformation of the estimates to the underlying scale resulted in heritability estimates from 0.12 to 0.17. Posterior means of the heritability of litter size in the Bayesian approach with bi-variate threshold models varied from 0.05 to 0.18. REML estimates of the genetic correlations between the two types of litter size ranged from 0.57 to 0.64 in the Suffolk and from 0.75 to 0.81 in the Texel. The posterior means of the genetic correlation (Bayesian analysis) were 0.40 and 0.44 for the Suffolk and 0.56 and 0.75 for the Texel in the sire and animal model respectively. A bivariate threshold model seems appropriate for the genetic evaluation of prolificacy in the breeds concerned. PMID- 15339633 TI - Bayesian segregation analysis of milk flow in Swiss dairy cattle using Gibbs sampling. AB - Segregation analyses with Gibbs sampling were applied to investigate the mode of inheritance and to estimate the genetic parameters of milk flow of Swiss dairy cattle. The data consisted of 204,397, 655,989 and 40,242 lactation records of milk flow in Brown Swiss, Simmental and Holstein cattle, respectively (4 to 22 years). Separate genetic analyses of first and multiple lactations were carried out for each breed. The results show that genetic parameters especially polygenic variance and heritability of milk flow in the first lactation were very similar under both mixed inheritance (polygenes + major gene) and polygenic models. Segregation analyses yielded very low major gene variances which favour the polygenic determinism of milk flow. Heritabilities and repeatabilities of milk flow in both Brown Swiss and Simmental were high (0.44 to 0.48 and 0.54 to 0.59, respectively). The heritability of milk flow based on scores of milking ability in Holstein was intermediate (0.25). Variance components and heritabilities in the first lactation were slightly larger than those estimates for multiple lactations. The results suggest that milk flow (the quantity of milk per minute of milking) is a relevant measurement to characterise the cows milking ability which is a good candidate trait to be evaluated for a possible inclusion in the selection objectives in dairy cattle. PMID- 15339634 TI - Identification and reciprocal introgression of a QTL affecting body mass in mice. AB - The aim of this study was to examine the effects of a QTL in different genetic backgrounds. A QTL affecting body mass on chromosome 6 was identified in an F(2) cross between two lines of mice that have been divergently selected for this trait. The effect of the QTL on mass increased between 6 and 10 weeks of age and was not sex-specific. Body composition analysis showed effects on fat-free dry body mass and fat mass. To examine the effect of this QTL in different genetic backgrounds, the high body mass sixth chromosome was introgressed into the low body mass genetic background and vice versa by repeated marker-assisted backcrossing. After three generations of backcrossing, new F(2) populations were established within each of the introgression lines by crossing individuals that were heterozygous across the sixth chromosome. The estimated additive effect of the QTL on 10-week body mass was similar in both genetic backgrounds and in the original F(2) population (i.e., approximately 0.4 phenotypic standard deviations); no evidence of epistatic interaction with the genetic background was found. The 95% confidence interval for the location of the QTL was refined to a region of approximately 7 cM between D6Mit268 and D6Mit123. PMID- 15339635 TI - Assignment of CPS1, OTC, CRYD2, ARG2 and ASS genes to the chicken RH map. AB - An attempt was made to assign five genes, CPS1, OTC, ASS, CRYD2, and ARG2, to chicken chromosomes (GGA) by radiation-hybrid mapping. OTC was assigned to GGA1; ARG2 to GGA5; CPS1 to GGA7; and CRYD2 to GGA19. ASS was not, however, assigned to a specific chromosomal position. PMID- 15339636 TI - Effect of (13)C-, (18)O- and (2)H-labeling on the infrared modes of UV-induced phenoxyl radicals. AB - The structure and environment of redox active tyrosines present in several metalloenzymes can be studied by resonance Raman spectroscopy or Fourier transform infrared difference spectroscopy. Assignments of the vibrational modes in vivo often requires in vitro studies on model compounds. This approach is briefly reviewed. New results are shown on the influence of isotope-labeling on the infrared spectra of tyrosine, [Formula: see text] and phenol radicals obtained in vitro by UV-irradiation. The infrared spectra of the radicals are dominated by the [Formula: see text] mode at 1515-1504 cm(-1). The frequency shifts induced on this mode by (13)C- (2)H-, and (18)O-labeling are reported. PMID- 15339637 TI - Fulfilling the promise of the cognitive neurosciences. AB - The paper by Meyer-Lindenberg and colleagues in this issue of Neuron provides strong evidence that the absence of one or more genes in Williams syndrome leads to highly circumscribed pathology in the dorsal visual stream. This program of research demonstrates that neurocognitive architecture follows the same principles in typical and atypical development. PMID- 15339638 TI - Food for thought: essential fatty acid protects against neuronal deficits in transgenic mouse model of AD. AB - Interactions between environmental and genetic factors may contribute to neurodegenerative disease. In this issue of Neuron, Calon et al. report that a diet low in an essential omega-3 polyunsaturated fatty acid (docosahexaenoic acid) depletes postsynaptic proteins and exacerbates behavioral alterations in a transgenic mouse model of Alzheimer's disease. PMID- 15339639 TI - Calcium waves rule and divide radial glia. AB - Radial glial proliferation is a critical step in the construction of cerebral cortex. In this issue of Neuron, Weissman and colleagues use time-lapse calcium imaging techniques to demonstrate that spontaneous calcium waves sweeping through cohorts of radial glia in the ventricular zone can modulate their proliferation during cerebral cortical development. PMID- 15339640 TI - How early is firing required for wiring? AB - Activity is known to be important for the refinement of neural connections in the developing brain. In this issue of Neuron, Hanson and Landmesser provide evidence that GABA-dependent spontaneous bursting of motor neurons in the embryonic spinal cord is required for the correct execution of an early axon pathfinding decision. PMID- 15339641 TI - Resetting the clock: Dexras1 defines a path. AB - The signaling pathways by which light and activity shift the circadian clock are not well understood. In this issue of Neuron, Cheng et al. analyze mice lacking Dexras1 (a Ras family GTPase protein) and demonstrate an important role for G(i/o) signaling mediating both photic and nonphotic phase shifts of the circadian clock. PMID- 15339642 TI - Clearance of Alzheimer's Abeta peptide: the many roads to perdition. AB - The amyloid hypothesis of Alzheimer's disease (AD) maintains that the accumulation of the amyloid beta protein (Abeta) is a critical event in disease pathogenesis. A great deal of both academic and commercial research has focused on the mechanisms by which Abeta is generated. However, investigations into the mechanisms underlying Abeta clearance have blossomed over the last several years. This minireview will summarize pathways involved in the removal of cerebral Abeta, including enzymatic degradation and receptor-mediated efflux out of the brain. PMID- 15339643 TI - Synaptic connectivity and neuronal morphology: two sides of the same coin. AB - Neurons often possess elaborate axonal and dendritic arbors. Why do these arbors exist and what determines their form and dimensions? To answer these questions, I consider the wiring up of a large highly interconnected neuronal network, such as the cortical column. Implementation of such a network in the allotted volume requires all the salient features of neuronal morphology: the existence of branching dendrites and axons and the presence of dendritic spines. Therefore, the requirement of high interconnectivity is, in itself, sufficient to account for the existence of these features. Moreover, the actual lengths of axons and dendrites are close to the smallest possible length for a given interconnectivity, arguing that high interconnectivity is essential for cortical function. PMID- 15339644 TI - In memoriam: Francis H.C. Crick. PMID- 15339645 TI - Neural basis of genetically determined visuospatial construction deficit in Williams syndrome. AB - A unique opportunity to understand genetic determinants of cognition is offered by Williams syndrome (WS), a well-characterized hemideletion on chromosome 7q11.23 that causes extreme, specific weakness in visuospatial construction (the ability to visualize an object as a set of parts or construct a replica). Using multimodal neuroimaging, we identified a neural mechanism underlying the WS visuoconstructive deficit. Hierarchical assessment of visual processing with fMRI showed isolated hypoactivation in WS in the parietal portion of the dorsal stream. In the immediately adjacent parietooccipital/intraparietal sulcus, structural neuroimaging showed a gray matter volume reduction in participants with WS. Path analysis demonstrated that the functional abnormalities could be attributed to impaired input from this structurally altered region. Our observations confirm a longstanding hypothesis about dorsal stream dysfunction in WS, demonstrate effects of a localized abnormality on visual information processing in humans, and define a systems-level phenotype for mapping genetic determinants of visuoconstructive function. PMID- 15339647 TI - Calcium waves propagate through radial glial cells and modulate proliferation in the developing neocortex. AB - The majority of neurons in the adult neocortex are produced embryonically during a brief but intense period of neuronal proliferation. The radial glial cell, a transient embryonic cell type known for its crucial role in neuronal migration, has recently been shown to function as a neuronal progenitor cell and appears to produce most cortical pyramidal neurons. Radial glial cell modulation could thus affect neuron production, neuronal migration, and overall cortical architecture; however, signaling mechanisms among radial glia have not been studied directly. We demonstrate here that calcium waves propagate through radial glial cells in the proliferative cortical ventricular zone (VZ). Radial glial calcium waves occur spontaneously and require connexin hemichannels, P2Y1 ATP receptors, and intracellular IP3-mediated calcium release. Furthermore, we show that wave disruption decreases VZ proliferation during the peak of embryonic neurogenesis. Taken together, these results demonstrate a radial glial signaling mechanism that may regulate cortical neuronal production. PMID- 15339646 TI - Docosahexaenoic acid protects from dendritic pathology in an Alzheimer's disease mouse model. AB - Learning and memory depend on dendritic spine actin assembly and docosahexaenoic acid (DHA), an essential n-3 (omega-3) polyunsaturated fatty acid (PFA). High DHA consumption is associated with reduced Alzheimer's disease (AD) risk, yet mechanisms and therapeutic potential remain elusive. Here, we report that reduction of dietary n-3 PFA in an AD mouse model resulted in 80%-90% losses of the p85alpha subunit of phosphatidylinositol 3-kinase and the postsynaptic actin regulating protein drebrin, as in AD brain. The loss of postsynaptic proteins was associated with increased oxidation, without concomitant neuron or presynaptic protein loss. n-3 PFA depletion increased caspase-cleaved actin, which was localized in dendrites ultrastructurally. Treatment of n-3 PFA-restricted mice with DHA protected against these effects and behavioral deficits and increased antiapoptotic BAD phosphorylation. Since n-3 PFAs are essential for p85-mediated CNS insulin signaling and selective protection of postsynaptic proteins, these findings have implications for neurodegenerative diseases where synaptic loss is critical, especially AD. PMID- 15339648 TI - Transmembrane/juxtamembrane domain-dependent Dscam distribution and function during mushroom body neuronal morphogenesis. AB - Besides 19,008 possible ectodomains, Drosophila Dscam contains two alternative transmembrane/juxtamembrane segments, respectively, derived from exon 17.1 and exon 17.2. We wondered whether specific Dscam isoforms mediate formation and segregation of axonal branches in the Drosophila mushroom bodies (MBs). Removal of various subsets of the 12 exon 4s does not affect MB neuronal morphogenesis, while expression of a Dscam transgene only partially rescues Dscam mutant phenotypes. Interestingly, differential rescuing effects are observed between two Dscam transgenes that each possesses one of the two possible exon 17s. Axon bifurcation/segregation abnormalities are better rescued by the exon 17.2 containing transgene, but coexpression of both transgenes is required for rescuing mutant viability. Meanwhile, exon 17.1 targets ectopically expressed Dscam-GFP to dendrites while Dscam[exon 17.2]-GFP is enriched in axons; only Dscam[exon 17.2] affects MB axons. These results suggest that exon 17.1 is minimally involved in axonal morphogenesis and that morphogenesis of MB axons probably involves multiple distinct exon 17.2-containing Dscam isoforms. PMID- 15339649 TI - Analysis of Dscam diversity in regulating axon guidance in Drosophila mushroom bodies. AB - Dscam is an immunoglobulin (Ig) superfamily member that regulates axon guidance and targeting in Drosophila. Alternative splicing potentially generates 38,016 isoforms differing in their extracellular Ig and transmembrane domains. We demonstrate that Dscam mediates the sorting of axons in the developing mushroom body (MB). This correlates with the precise spatiotemporal pattern of Dscam protein expression. We demonstrate that MB neurons express different arrays of Dscam isoforms and that single MB neurons express multiple isoforms. Two different Dscam isoforms differing in their extracellular domains introduced as transgenes into single mutant cells partially rescued the mutant phenotype. Expression of one isoform of Dscam in a cohort of MB neurons induced dominant phenotypes, while expression of a single isoform in a single cell did not. We propose that different extracellular domains of Dscam share a common function and that differences in isoforms expressed on the surface of neighboring axons influence interactions between them. PMID- 15339650 TI - Normal patterns of spontaneous activity are required for correct motor axon guidance and the expression of specific guidance molecules. AB - Rhythmic spontaneous electrical activity occurs in many parts of the developing nervous system, where it plays essential roles in the refinement of neural connections. By blocking or slowing this bursting activity, via in ovo drug applications at precise developmental periods, we show that such activity is also required at much earlier stages for spinal motoneurons to accurately execute their first major dorsal-ventral pathfinding decision. Blockade or slowing of rhythmic bursting activity also prevents the normal expression patterns of EphA4 and polysialic acid on NCAM, which may contribute to the pathfinding errors observed. More prolonged (E2-5) blockade resulted in a downregulation of LIM homeodomain transcription factors, but since this occurred only after the pathfinding errors and alterations in guidance molecules, it cannot have contributed to them. PMID- 15339651 TI - Or83b encodes a broadly expressed odorant receptor essential for Drosophila olfaction. AB - Fruit flies are attracted by a diversity of odors that signal the presence of food, potential mates, or attractive egg-laying sites. Most Drosophila olfactory neurons express two types of odorant receptor genes: Or83b, a broadly expressed receptor of unknown function, and one or more members of a family of 61 selectively expressed receptors. While the conventional odorant receptors are highly divergent, Or83b is remarkably conserved between insect species. Two models could account for Or83b function: it could interact with specific odor stimuli independent of conventional odorant receptors, or it could act in concert with these receptors to mediate responses to all odors. Our results support the second model. Dendritic localization of conventional odorant receptors is abolished in Or83b mutants. Consistent with this cellular defect, the Or83b mutation disrupts behavioral and electrophysiological responses to many odorants. Or83b therefore encodes an atypical odorant receptor that plays an essential general role in olfaction. PMID- 15339652 TI - Dexras1 potentiates photic and suppresses nonphotic responses of the circadian clock. AB - Circadian rhythms of physiology and behavior are generated by biological clocks that are synchronized to the cyclic environment by photic or nonphotic cues. The interactions and integration of various entrainment pathways to the clock are poorly understood. Here, we show that the Ras-like G protein Dexras1 is a critical modulator of the responsiveness of the master clock to photic and nonphotic inputs. Genetic deletion of Dexras1 reduces photic entrainment by eliminating a pertussis-sensitive circadian response to NMDA. Mechanistically, Dexras1 couples NMDA and light input to Gi/o and ERK activation. In addition, the mutation greatly potentiates nonphotic responses to neuropeptide Y and unmasks a nonphotic response to arousal. Thus, Dexras1 modulates the responses of the master clock to photic and nonphotic stimuli in opposite directions. These results identify a signaling molecule that serves as a differential modulator of the gated photic and nonphotic input pathways to the circadian timekeeping system. PMID- 15339653 TI - Neuronal synchrony mediated by astrocytic glutamate through activation of extrasynaptic NMDA receptors. AB - Fast excitatory neurotransmission is mediated by activation of synaptic ionotropic glutamate receptors. In hippocampal slices, we report that stimulation of Schaffer collaterals evokes in CA1 neurons delayed inward currents with slow kinetics, in addition to fast excitatory postsynaptic currents. Similar slow events also occur spontaneously, can still be observed when neuronal activity and synaptic glutamate release are blocked, and are found to be mediated by glutamate released from astrocytes acting preferentially on extrasynaptic NMDA receptors. The slow currents can be triggered by stimuli that evoke Ca2+ oscillations in astrocytes, including photolysis of caged Ca2+ in single astrocytes. As revealed by paired recording and Ca2+ imaging, a striking feature of this NMDA receptor response is that it occurs synchronously in multiple CA1 neurons. Our results reveal a distinct mechanism for neuronal excitation and synchrony and highlight a functional link between astrocytic glutamate and extrasynaptic NMDA receptors. PMID- 15339654 TI - Optimal information storage and the distribution of synaptic weights: perceptron versus Purkinje cell. AB - It is widely believed that synaptic modifications underlie learning and memory. However, few studies have examined what can be deduced about the learning process from the distribution of synaptic weights. We analyze the perceptron, a prototypical feedforward neural network, and obtain the optimal synaptic weight distribution for a perceptron with excitatory synapses. It contains more than 50% silent synapses, and this fraction increases with storage reliability: silent synapses are therefore a necessary byproduct of optimizing learning and reliability. Exploiting the classical analogy between the perceptron and the cerebellar Purkinje cell, we fitted the optimal weight distribution to that measured for granule cell-Purkinje cell synapses. The two distributions agreed well, suggesting that the Purkinje cell can learn up to 5 kilobytes of information, in the form of 40,000 input-output associations. PMID- 15339655 TI - Centromere glue provides spindle cue. AB - During cell division, accurate distribution of the genome by the mitotic spindle requires that sister chromatids remain tethered until the right moment. Studies of an uncharacterized vertebrate protein, Sgo (Salic et al., 2004 [this issue of Cell]), reveal dual roles as a chromosome cohesion factor and a regulator of spindle microtubule dynamics. PMID- 15339656 TI - Epithelial stem cells: stepping out of their niche. AB - In this issue of Cell, have shown that two subpopulations of cells exist within the hair follicle stem cell niche. Despite being partially differentiated, clonal populations of suprabasal bulge region cells can regenerate skin and hair follicles as well as a new stem cell niche. The findings suggest that early lineage commitments of epithelial cells in the hair follicle may be reversible. PMID- 15339657 TI - Sculpting heart valves with NFATc and VEGF. AB - Heart valves are of vital importance for our moment-to-moment existence, but how they form remains a mystery. In this issue of Cell, Chang et al. reveal a novel role for calcineurin, NFATs, and VEGF in valve formation. Dynamic changes in NFAT/VEGF expression in regional myocardial and endocardial fields and developmental windows orchestrate this complex process. PMID- 15339658 TI - Wagging the dogma; tissue-specific cell cycle control in the mouse embryo. AB - The family of cyclin-dependent kinases (Cdks) lies at the core of the machinery that drives the cell division cycle. Studies in cultured mammalian cells have provided insight into the cellular functions of many Cdks. Recent Cdk and cyclin knockouts in the mouse show that the functions of G1 cell cycle regulatory genes are often essential only in specific cell types, pointing to our limited understanding of tissue-specific expression, redundancy, and compensating mechanisms in the Cdk network. PMID- 15339659 TI - The lingering enigma of the allelic exclusion mechanism. AB - B lymphocytes produce diverse antibody specificities by "randomly" assembling antibody genes from germline segments. Yet, though each B lymphocyte has multiple allelic loci for the different antibody chains, each clonally derived mature B lymphocyte expresses a single species of antibody with a unique specificity via a process termed allelic exclusion. Despite some progress, the precise mechanism of allelic exclusion remains an enigma. PMID- 15339660 TI - Histone deimination antagonizes arginine methylation. AB - Methylation of arginine residues within histone H3 has been linked to active transcription. This modification appears on the estrogen-regulated pS2 promoter when the CARM1 methyltransferase is recruited during transcriptional activation. Here we describe a process, deimination, that converts histone arginine to citrulline and antagonizes arginine methylation. We show that peptidyl arginine deiminase 4 (PADI4) specifically deiminates, arginine residues R2, R8, R17, and R26 in the H3 tail. Deimination by PADI4 prevents arginine methylation by CARM1. Dimethylation of arginines prevents deimination by PADI4 although monomethylation still allows deimination to take place. In vivo targeting experiments on an endogenous promoter demonstrate that PADI4 can repress hormone receptor-mediated gene induction. Consistent with a repressive role for PADI4, this enzyme is recruited to the pS2 promoter following hormone induction when the gene is transcriptionally downregulated. The recruitment of PADI4 coincides with deimination of the histone H3 N-terminal tail. These results define deimination as a novel mechanism for antagonizing the transcriptional induction mediated by arginine methylation. PMID- 15339661 TI - Chromatin architecture of the human genome: gene-rich domains are enriched in open chromatin fibers. AB - We present an analysis of chromatin fiber structure across the human genome. Compact and open chromatin fiber structures were separated by sucrose sedimentation and their distributions analyzed by hybridization to metaphase chromosomes and genomic microarrays. We show that compact chromatin fibers originate from some sites of heterochromatin (C-bands), and G-bands (euchromatin). Open chromatin fibers correlate with regions of highest gene density, but not with gene expression since inactive genes can be in domains of open chromatin, and active genes in regions of low gene density can be embedded in compact chromatin fibers. Moreover, we show that chromatin fiber structure impacts on further levels of chromatin condensation. Regions of open chromatin fibers are cytologically decondensed and have a distinctive nuclear organization. We suggest that domains of open chromatin may create an environment that facilitates transcriptional activation and could provide an evolutionary constraint to maintain clusters of genes together along chromosomes. PMID- 15339662 TI - Vertebrate shugoshin links sister centromere cohesion and kinetochore microtubule stability in mitosis. AB - Drosophila MEI-S332 and fungal Sgo1 genes are essential for sister centromere cohesion in meiosis I. We demonstrate that the related vertebrate Sgo localizes to kinetochores and is required to prevent premature sister centromere separation in mitosis, thus providing an explanation for the differential cohesion observed between the arms and the centromeres of mitotic sister chromatids. Sgo is degraded by the anaphase-promoting complex, allowing the separation of sister centromeres in anaphase. Intriguingly, we show that Sgo interacts strongly with microtubules in vitro and that it regulates kinetochore microtubule stability in vivo, consistent with a direct microtubule interaction. Sgo is thus critical for mitotic progression and chromosome segregation and provides an unexpected link between sister centromere cohesion and microtubule interactions at kinetochores. PMID- 15339663 TI - Cytokinesis monitoring during development; rapid pole-to-pole shuttling of a signaling protein by localized kinase and phosphatase in Caulobacter. AB - For successful generation of different cell types by asymmetric cell division, cell differentiation should be initiated only after completion of division. Here, we describe a control mechanism by which Caulobacter couples the initiation of a developmental program to the completion of cytokinesis. Genetic evidence indicates that localization of the signaling protein DivK at the flagellated pole prevents premature initiation of development. Photobleaching and FRET experiments show that polar localization of DivK is dynamic with rapid pole-to-pole shuttling of diffusible DivK generated by the localized activities of PleC phosphatase and DivJ kinase at opposite poles. This shuttling is interrupted upon completion of cytokinesis by the segregation of PleC and DivJ to different daughter cells, resulting in disruption of DivK localization at the flagellated pole and subsequent initiation of development in the flagellated progeny. Thus, dynamic polar localization of a diffusible protein provides a control mechanism that monitors cytokinesis to regulate development. PMID- 15339664 TI - Endocytosis by random initiation and stabilization of clathrin-coated pits. AB - Clathrin-coated vesicles carry traffic from the plasma membrane to endosomes. We report here the real-time visualization of cargo sorting and endocytosis by clathrin-coated pits in living cells. We have detected the formation of coats by monitoring incorporation of fluorescently tagged clathrin or its adaptor AP-2; we have also followed clathrin-mediated uptake of transferrin and of single LDL or reovirus particles. The intensity of a cargo-loaded clathrin cluster grows steadily during its lifetime, and the time required to complete assembly is proportional to the size of the cargo particle. These results are consistent with a nucleation-growth mechanism and an approximately constant growth rate. There are no strongly preferred nucleation sites. A proportion of the nucleation events are weak and short lived. Cargo incorporation occurs primarily or exclusively in a newly formed coated pit. Our data lead to a model in which coated pits initiate randomly but collapse unless stabilized, perhaps by cargo capture. PMID- 15339665 TI - Structure, exchange determinants, and family-wide rab specificity of the tandem helical bundle and Vps9 domains of Rabex-5. AB - The Rab5 GTPase, an essential regulator of endocytosis and endosome biogenesis, is activated by guanine-nucleotide exchange factors (GEFs) that contain a Vps9 domain. Here, we show that the catalytic core of the Rab GEF Rabex-5 has a tandem architecture consisting of a Vps9 domain stabilized by an indispensable helical bundle. A family-wide analysis of Rab specificity demonstrates high selectivity for Rab5 subfamily GTPases. Conserved exchange determinants map to a common surface of the Vps9 domain, which recognizes invariant aromatic residues in the switch regions of Rab GTPases and selects for the Rab5 subfamily by requiring a small nonacidic residue preceding a critical phenylalanine in the switch I region. These and other observations reveal unexpected similarity with the Arf exchange site in the Sec7 domain. PMID- 15339666 TI - Alternative splicing of Drosophila Dscam generates axon guidance receptors that exhibit isoform-specific homophilic binding. AB - Dscam is an immunoglobulin (Ig) superfamily protein required for the formation of neuronal connections in Drosophila. Through alternative splicing, Dscam potentially gives rise to 19,008 different extracellular domains linked to one of two alternative transmembrane segments, resulting in 38,016 isoforms. All isoforms share the same domain structure but contain variable amino acid sequences within three Ig domains in the extracellular region. We demonstrate that different isoforms exhibit different binding specificity. Each isoform binds to itself but does not bind or binds poorly to other isoforms. The amino acid sequences of all three variable Ig domains determine binding specificity. Even closely related isoforms sharing nearly identical amino acid sequences exhibit isoform-specific binding. We propose that this preferential homophilic binding specificity regulates interactions between cells and contributes to the formation of complex patterns of neuronal connections. PMID- 15339667 TI - Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche. AB - In adult skin, each hair follicle contains a reservoir of stem cells (the bulge), which can be mobilized to regenerate the new follicle with each hair cycle and to reepithelialize epidermis during wound repair. Here we report new methods that permit their clonal analyses and engraftment and demonstrate the two defining features of stem cells, namely self-renewal and multipotency. We also show that, within the bulge, there are two distinct populations, one of which maintains basal lamina contact and temporally precedes the other, which is suprabasal and arises only after the start of the first postnatal hair cycle. This spatial distinction endows them with discrete transcriptional programs, but surprisingly, both populations are growth inhibited in the niche but can self-renew in vitro and make epidermis and hair when grafted. These findings suggest that the niche microenvironment imposes intrinsic "stemness" features without restricting the establishment of epithelial polarity and changes in gene expression. PMID- 15339668 TI - A field of myocardial-endocardial NFAT signaling underlies heart valve morphogenesis. AB - The delicate leaflets that make up vertebrate heart valves are essential for our moment-to-moment existence. Abnormalities of valve formation are the most common serious human congenital defect. Despite their importance, relatively little is known about valve development. We show that the initiation of heart valve morphogenesis in mice requires calcineurin/NFAT to repress VEGF expression in the myocardium underlying the site of prospective valve formation. This repression of VEGF at E9 is essential for endocardial cells to transform into mesenchymal cells. Later, at E11, a second wave of calcineurin/NFAT signaling is required in the endocardium, adjacent to the earlier myocardial site of NFAT action, to direct valvular elongation and refinement. Thus, NFAT signaling functions sequentially from myocardium to endocardium within a valvular morphogenetic field to initiate and perpetuate embryonic valve formation. This mechanism also operates in zebrafish, indicating a conserved role for calcineurin/NFAT signaling in vertebrate heart valve morphogenesis. PMID- 15339669 TI - Contemporary issues in the fight against blood doping in sport. PMID- 15339670 TI - Blood doping research in Haematologica. PMID- 15339671 TI - Diagnosis and treatment of essential thrombocythemia. PMID- 15339672 TI - No increase in age-specific incidence of myelodysplastic syndromes. AB - BACKGROUND AND OBJECTIVES: Epidemiological data on myelodysplastic syndromes (MDS) are sparse. However, the available evidence indicates that MDS is among the most common hematologic diseases. DESIGN AND METHODS: In a previous study, we showed that incidence rates rose from 1 to 4.1 in the period 1976-1990, with no increase between 1986 and 1990. We extended our study, covering the years 1991 through 2001. Diagnostic criteria and cytomorphology remained the same, consistently applying the criteria of the FAB classification. RESULTS: Three hundred and eight new cases of MDS were identified in the town district of Dusseldorf, which provides the reference population for calculating incidences. There was no further rise in MDS incidence. The crude incidence in the period 1991-2001 was 4.9 (5.52 in males, 4.36 in females), which is similar to that in the 1986-1990 period. Age specific incidences were 8.7, 24.5, and 31.3 for the age groups 60-70, 71-80, and 80-90, respectively. The incidence of MDS in the age group >70 years was significantly higher among males (42.3) than among females (19.0). The preponderance of males was found among patients with refractory anemia with excess blasts (16 vs 3), refractory anemia with excess blasts in transformation (7 vs 2), refractory anemia (15 vs 6) and chronic myelomonocytic leukemia (7 vs 2), whereas, somewhat surprisingly, the age-related incidences refractory anemia with ringed sideroblasts was similar for males and females (4 vs 5). The data on MDS incidence in the town district of Dusseldorf showed a plateau since 1986. INTERPRETATION AND CONCLUSIONS: Interpreting our data covering a 26-year period, we feel that the increase in the early years primarily reflected improved case ascertainment, whereas our new data may provide a good approximation of the true incidence of MDS. There is no evidence that the age adjusted incidence of MDS is rising. PMID- 15339673 TI - Value of bone marrow biopsy in the diagnosis of essential thrombocythemia. AB - BACKGROUND AND OBJECTIVES: Essential thrombocythemia (ET) is a Philadelphia chromosome-negative chronic myeloproliferative disorder (CMPD) whose diagnosis, according to the Polycythemia Vera Study Group (PVSG) criteria, does not include histopathological data. The new WHO classification of CMPD has supplied new diagnostic guidelines which highlight the value of histopathology and facilitate a more precise differentiation of ET from reactive conditions and other CMPDs. DESIGN AND METHODS: Bone marrow biopsies from 142 adult patients diagnosed with ET according to PVSG criteria were evaluated using the new WHO classification. Megakaryocyte morphology and arrangement, amount of fibrosis and a clustering index were studied along with determination of microvessel density (MVD), amount of CD34+ cells and percentage of MIB-1+ cells and megakaryocytes. The last value, indicated as megakaryocyte proliferation index (MPI), was determined and expressed as a percentage of the counted cells. RESULTS: According to WHO criteria the 142 biopsies were classified as follows: ET (21%); Idiopathic myelofibrosis (IMF) grade 0 (30%); IMF-1 (34%); IMF-2 (10%) ET/IMF-0 (5%). A significant difference (p<0.001) was observed between clustering index values in ET and IMF cases. A peculiar proliferative feature of megakaryocytes, defined coupling, was detected in all ET cases. MVD was more pronounced and the number of CD34+ cells higher in cases of IMF than in cases of ET (p <0.005; p = 0.001, respectively) and MVD significantly correlated with the extent of fibrosis (r=0.861). ET cases showed the lowest values of proliferation; IMF-0 and IMF-1 showed higher values while a decrease of MPI was observed in IMF-2 in accordance with the increase of fibrosis. INTERPRETATION AND CONCLUSIONS: In the diagnosis of thrombocythemic disorders, a multidisciplinary approach must include the evaluation of bone marrow biopsies. Some histopathological criteria, along with the use of markers related to activity and proliferation such as CD34 and MIB-1, underline the biological differences between ET and prefibrotic states of IMF. PMID- 15339674 TI - KIT activating mutations: incidence in adult and pediatric acute myeloid leukemia, and identification of an internal tandem duplication. AB - BACKGROUND AND OBJECTIVES: Mutations of KIT receptor tyrosine kinase are involved in the constitutive activation and development of human hematologic malignancies. Gain-of-function mutations in the second intracellular kinase domain (TK2) and in the juxtamembrane domain are described in patients with core binding factor acute myeloid leukemia (CBFL) and are associated with leukocytosis. We evaluated the incidence of KIT mutation in 52 adult patients with de novo CBFL and in 49 FLT3/ITD-negative childhood patients with de novo acute myeloid leukemia (AML), excluding cases of acute promyelocytic leukemia. DESIGN AND METHODS: In order to analyze the role of KIT in CBFL we examined the KIT mutations in 52 adult CBFL, including 15 previously reported patients, and in 49 non-APL childhood AML patients using sensitive detection methods. We correlated our findings with the presence of trisomy 4 and investigated the relationship of the extra chromosome 4 with KIT mutations. RESULTS: Several kinds of gain-of-function KIT mutations were found in 24 of the 52 (46.1%) adult CBFL cases and 6 of the 49 (12.2%) non-APL childhood AML patients. KIT mutations were detected in 4 of the 8 adult patients and one childhood AML case bearing trisomy of chromosome 4 as either the sole cytogenetic aberration or a karyotypic aberration additional to t(8;21). In three of the trisomy 4 cases we demonstrated that trisomy 4 leads to duplication of the KIT mutated allele. INTERPRETATION AND CONCLUSIONS: These results underline that the KIT gene is activated in AML characterized by distinct cytogenetic and molecular genetic patterns and represents the most frequently mutated target in adult CBFL. PMID- 15339675 TI - Prognostic implications of Wilms' tumor gene (WT1) expression in patients with de novo acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: The Wilms' tumor (WT1) gene is overexpressed in patients with most forms of acute leukemia. Several studies have reported the usefulness of quantitative assessment of WT1 expression as a molecular marker of minimal residual disease. However, the biological significance and the prognostic impact of WT1 overexpression in acute myeloid leukemia (AML) is still uncertain. DESIGN AND METHODS: We analyzed the prognostic relevance of WT1 expression in a cohort of 77 adult patients with AML, using a real-time quantitative reverse transcription polymerase chain reaction approach. RESULTS: WT1 expression was significantly higher in AML patients than in normal controls (p = 0.0001). The normalized levels of WT1 with respect to the control gene for beta-glucuronidase (GUS) in AML samples showed a median WT1/GUS ratio of 0.93 (range 0-25). We classified the patients into two groups according to this ratio. Forty patients (52%) showed a WT1/GUS ratio 1. A ratio > 1, although significantly associated with FLT3 mutations, was the strongest independent prognostic factor for disease-free survival (p = 0.004), relapse risk (p = 0.005) and cumulative incidence risk (p = 0.01). This adverse prognostic value was more evident in patients aged 60 years and younger. INTERPRETATION AND CONCLUSIONS: The WT1/GUS ratio is an independent prognostic factor for predicting relapse in patients with AML and it could be included as part of the initial evaluation to establish more defined risk groups. PMID- 15339676 TI - Combined analysis of bcl-2 and MDR1 proteins in 256 cases of acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: The objective of this study was to investigate the expression of MDR1 and bcl-2 proteins in de novo acute myeloid leukemia (AML). DESIGN AND METHODS: The expression of MDR1 and bcl-2 was analyzed by flow cytometry in a large series of 256 consecutive cases of AML. The results were recorded as percentage of positivity and relative mean fluorescence intensity (rMFI). To determine individual protein levels, an index which equals the product of the percentage of positive cells and rMFI was generated. RESULTS: Using cut offs of >or=800 and 300 of the index value for bcl-2 and MDR1 expression, respectively, we identified 4 different classes of AML: 1) double negative; 2) single positive bcl-2+/MDR1-; 3) single positive bcl-2-/MDR1+; 4) double positive. The highest incidence of double negative cases was observed in the M2 class whereas double positive cases occurred more frequently in the M4, M5 and M6 subgroups. Seventy-eight percent and 71% of M0 and M1, respectively, showed single positive bcl-2+/MDR1- expression (p = 0.00001). Twenty-eight percent of patients belonging to the double positive category achieved complete remission, whereas for double negative, single positive bcl-2+MDR1- and single positive bcl 2-/MDR1+ category, the complete remission rate was 69%, 52% and 56%, respectively (p = 0.00038). In multivariate analysis, the double positive status independently affected frequency of complete remission (p = 0.008). INTERPRETATION AND CONCLUSIONS: Bcl-2 is over-expressed in CD34+ AML; conversely, MDR1 is over expressed in CD34- AML. However, the combined expression of the two proteins defines a subset of AML with a very poor prognosis. PMID- 15339677 TI - Treatment of primary acute myeloid leukemia: results of a prospective multicenter trial including high-dose cytarabine or stem cell transplantation as post remission strategy. AB - BACKGROUND AND OBJECTIVES: To evaluate a regimen of induction and consolidation chemotherapy, followed by a post-remission therapy which depended on age and cytogenetics, in patients with primary acute myeloid leukemia. DESIGN AND METHODS: Two hundred patients up to 60 years old received idarubicin, standard dose cytarabine and etoposide as induction chemotherapy and one consolidation course including intermediate dose cytarabine and mitoxantrone. Subsequently, patients with favorable cytogenetics, [i.e., t(8;21), inv(16)] were scheduled to receive 2 courses of high-dose cytarabine. The remainder were scheduled for allogeneic stem cell transplantation (SCT), if 50 years old or lacking a donor. RESULTS: In patients with favorable cytogenetics the 4-year probabilities of survival and leukemia-free survival (LFS) were 62+/-9% and 41+/-10%, respectively. The results were better in patients with t(8;21). LFS at 4 years in patients 50 years old assigned to auto-SCT had a 4-year LFS of 17+/-9%. Adverse cytogenetics and white blood cell count >or= 20 yen 109/L at diagnosis were associated with lower probability of survival and leukemia-free survival. INTERPRETATION AND CONCLUSIONS: We confirmed that high dose cytarabine seems a good option for patients with t(8;21). Autologous and allogeneic SCT led to similar leukemia-free survival in patients 65 years) with aggressive non-Hodgkin's lymphoma (NHL) treated with chemotherapy regimens containing anthracyclines. DESIGN AND METHODS: QOL was evaluated in a population of elderly patients with aggressive NHL enrolled in a phase III clinical trial run by the Intergruppo Italiano Linfomi (IIL) from 1996 to 1999 to compare two different anthracycline containing regimens (mini-CEOP vs P-VEBEC). The EORTC-QLQ-C30 questionnaire, which has already been validated in oncology, was used. The questionnaire was administered at the time of diagnosis, half way through the chemotherapy and at the time of restaging. RESULTS: Ninety-one patients completed pre-therapy and post-therapy questionnaires and they are the subject of this report. Baseline QOL assessment showed a strong correlation of poor values of QOL with anemia and high risk according to the International Prognostic Index (IPI). At the end of treatment no functional scales showed worse values. A significant improvement was observed for pain (p=0.003), appetite (p=0.006), sleep (p=0.015) and global health (p=0.027). Considering only the 50 patients who achieved a complete remission (CR), an improvement was also recorded for emotional state (p=0.10), role (p=0.05), constipation (p=0.04) and global QOL (p=0.05). INTERPRETATION AND CONCLUSIONS: The EORTC-QLQ-C30 is feasible even in a population of elderly patients, in whom it had never been tested before. The improvement of QOL at the end of the treatment demonstrated that the symptoms of the disease have a greater negative influence on the patient's life than do the side effects of the therapy. PMID- 15339682 TI - Molecular characterization of a factor VII deficient patient supports the importance of the second epidermal growth factor-like domain. AB - BACKGROUND AND OBJECTIVES: Although a large number of gene mutations have been characterized in patients with factor VII (FVII) deficiency, few naturally occurring mutations have been described in epidermal growth factor (EGF)-like domains. We investigated a 6-year old Italian girl who had low functional and antigenic FVII plasma levels. DESIGN AND METHODS: Plasma levels of FVII activity and antigen were evaluated in the propositus and her relatives. Mutation screening was performed by sequencing the FVII gene. The effect of the identified FVII mutations was investigated by protein expression in transfected cells. RESULTS: The propositus was shown to be a compound heterozygote for a known (Arg110Cys) and a novel (Asp123Tyr) missense mutation both occurring in the second EGF-like domain. In transfected cells, expression of the Arg110Cys mutation reduced the amount of intracellular and secreted FVII protein (48% and 18%, respectively). Likewise, cells transfected with the Asp123Tyr mutation gave rise to low intracellular (40%) and extracellular (4%) FVII antigen levels. In conditioned media, FVII procoagulant activity was reduced accordingly (10% and <1%, respectively). INTERPRETATION AND CONCLUSIONS: Transient expression of the identified FVII mutations caused severely reduced but detectable FVII antigen and activity levels. The present findings suggest that the two naturally occurring missense mutations identified within the second EGF-like domain severely affect FVII protein handling, impairing the correct folding of FVII. PMID- 15339683 TI - Anti-tissue factor pathway inhibitor activity in subjects with antiphospholipid syndrome is associated with increased thrombin generation. AB - BACKGROUND AND OBJECTIVES: Immunoglobulin G (IgG) fractions from subjects with antiphospholipid syndrome (aPS) have previously been demonstrated to have inhibitory activity against tissue factor pathway inhibitor (TFPI). This may contribute to the development of a prothrombotic state by impaired regulation of the tissue factor (TF) pathway. This study investigated the effect that IgG fractions from aPS subjects containing anti-TFPI activity have on in vitro TF induced thrombin generation. DESIGN AND METHODS: TFPI and anti-TFPI activities were determined in normal controls (n=29) and aPS subjects (n=57). TFPI activity was determined using an amidolytic assay based on the generation of factor Xa. Anti-TFPI activity was determined after incubating IgG isolated from a control or subject plasma with pooled normal plasma, using the TFPI activity assay. The influence of IgG fractions from controls (n=10) and subjects (n=23) on TF-induced in vitro thrombin generation was determined using a chromogenic assay of thrombin activity. RESULTS: TFPI activity in controls (1.13+/-0.25 U/mL) was significantly lower than in subjects (1.30+/-0.42 U/mL) (p < 0.05). Anti-TFPI activity was significantly higher in subjects than controls (p = 0.0001). TF-induced thrombin generation was positively associated with anti-TFPI activity (r = 0.356; p > 0.05), with increased levels of each demonstrated in 5 subjects. INTERPRETATION AND CONCLUSIONS: Anti-TFPI activity was confirmed in 65% of aPS subjects. IgG fractions demonstrated a variable ability to interfere with TFPI function and TF induced thrombin generation. Cross-reacting antiphospholipid antibodies and/or other entities may interfere with TFPI function, resulting in a net increase in thrombin generation and an increased thrombotic risk. PMID- 15339684 TI - Analysis of human reticulocyte genes reveals altered erythropoiesis: potential use to detect recombinant human erythropoietin doping. AB - BACKGROUND AND OBJECTIVES: Enhancement of oxygen delivery to tissues is associated with improved sporting performance. One way of enhancing oxygen delivery is to take recombinant human erythropoietin (rHuEpo), which is an unethical and potentially dangerous practice. However, detection of the use of rHuEpo remains difficult in situations such as: i) several days after the end of treatment ii) when a treatment with low doses is conducted iii) if the rHuEpo effect is increased by other substances. In an attempt to detect rHuEpo abuse, we selected erythroid gene markers from a SAGE library and analyzed the effects of rHuEpo administration on expression of the HBB, FTL and OAZ genes. DESIGN AND METHODS: Ten athletes were assigned to the rHuEpo or placebo group. The rHuEpo group received subcutaneous injections of rHuEpo (50 UI/kg three times a week, 4 weeks; 20 UI/kg three times a week, 2 weeks). HBB, FTL and OAZ gene profiles were monitored by real time-polymerase chain reaction (PCR) quantification during and for 3 weeks after drug administration. RESULTS: The global analysis of these targeted genes detected in whole blood samples showed a characteristic profile of subjects misusing rHuEpo with a increase above the threshold levels. The individual analysis of OAZ mRNA seemed indicative of rHuEpo treatment. INTERPRETATION AND CONCLUSIONS: The performance-enhancing effect of rHuEpo treatment is greater than the duration of hematologic changes associated with rHuEpo misuse. Although direct electrophoretic methods to detect rHuEpo have been developed, recombinant isoforms of rHuEpo are not detectable some days after the last subcutaneous injection. To overcome these limitations indirect OFF models have been developed. Our data suggest that, in the near future, it will be possible to consolidate results achievable with the OFF models by analyzing selected erythroid gene markers as a supplement to indirect methods. PMID- 15339685 TI - Prognostic factors and therapeutic options for relapsed or refractory acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: Considerable progress has been made in the treatment of acute myeloid leukemia (AML); however, current therapeutic results are still unsatisfactory in untreated patients and poorer in those with primary refractory or relapsed disease. The biological and clinical characteristics of relapsed AML are analyzed here. DESIGN AND METHODS: Most relevant literature in English language on relapsed AML from 1990 to 2004 was considered paying particular attention to the heterogeneity of the disease and prognostic factors at the time of relapse; therapeutic results in terms of second complete remission (CR) with conventional chemotherapy, stem cell transplantation and new agents are also summarized. RESULTS: Molecular relapse is a current indication for treatment of acute promyelocytic leukemia (APL); however, data are emerging for the treatment of molecular relapse in AML other than APL, such as AML with t(8;21) and AML with inv(16). Age, duration of first remission and cytogenetics are the most relevant prognostic factors in relapsed AML. Promising therapeutic results have been reported for the antiCD33 monoclonal antibody conjugated with calicheamicin and the new nucleoside analog clofarabine; preliminary studies indicate that FLT3, farnesyl-transferase and bcl-2 inhibitors are active in relapsed AML. CONCLUSIONS: All relapsed elderly patients and young adults with CR1 lasting for less than 12 months are ideal candidates for experimental therapies. Efficiently conducted phase II randomized trials are needed in order to achieve relevant information to be translated into phase III trials. PMID- 15339686 TI - Genetic studies suggest a novel Portuguese origin for hemoglobin Porto Alegre. AB - Molecular studies performed in Portuguese and Brazilian cases of hemoglobin Porto Alegre [beta9 Ser->Cys] revealed that the mutation is in association with the Mediterranean haplotype I and framework 1 and that it is also in cis with an undescribed intragenic polymorphism (codon 27, GCC->GCT). Based upon these findings, and reinforced by historical data, we suggest that hemoglobin Porto Alegre originated from a single mutational event in the Portuguese population and was then spread to South America, namely to Brazil. PMID- 15339687 TI - Simultaneous detection of alpha-thalassemia and beta-thalassemia by oligonucleotide microarray. AB - In this study, we describe a reliable microarray-based assay for the simultaneous detection of alpha/beta-globin genotypes. The efficiency and specificity of this method were evaluated by blinded analysis of 1,880 samples. The assay provides unambiguous detection of complex combinations of heterozygous, compound heterozygous and homozygous alpha/beta-thalassemia genotypes. PMID- 15339688 TI - Lack of mutations in the human telomerase RNA component (hTERC) gene in Fanconi's anemia. AB - As some patients with Fanconi s anemia (FA) present excessive telomere shortening correlating with poor outcome, we investigated whether human telomerase RNA component (hTERC) mutations also play a role in telomere shortening in 115 FA patients. Only one patient was heterozygous for the G58A polymorphism. No other mutation or deletion was found. We conclude that hTERC gene mutations do not contribute to telomere shortening in FA. PMID- 15339689 TI - Molecular diagnosis of leukemic cerebrospinal fluid cells in children with newly diagnosed acute lymphoblastic leukemia. AB - Cytomorphology and IgH/T-cell receptor g clonal gene rearrangements detected by polymerase chain reaction (PCR) homo/heteroduplex analysis and direct sequencing were evaluated in cerebrospinal fluid (CSF) free of red-blood cells at diagnosis of 37 children with acute lymphoblastic leukemia. Molecular CSF involvement was greater as detected by molecular analysis than observed by morphologic criteria (45.9% vs 5.4%). The 4-year event-free survival was lower in the group with molecularly detected CSF involvement (p = 0.01). PMID- 15339690 TI - Effect of interleukin-1beta and glutathione S-transferase genotypes on the development of gastric mucosa-associated lymphoid tissue lymphoma. AB - e tested whether polymorphic variations in glutathione S-transferase genes (GSTM1, GSTT1, GSTP1) and interleukin-1 (IL-1beta and IL-1RN) genes confer susceptibility to mucosa-associated lymphoid tissue lymphomas (MALT) in a Chinese population. The rates of GSTM1, GSTP1, IL-1beta and IL-1RN genotypes did not differ between patients and controls. However, GSTT1 null genotypes were significantly more common in patients with MALT lymphomas (43/75 vs. 138/321, p=0.029; OR=1.8, 95% CI: 1.1-3.0) than in controls. Our results suggest that a glutathione S-transferase defect plays a role in MALT lymphoma. PMID- 15339691 TI - Treatment of heparin-induced thrombocytopenia with fondaparinux. AB - Anticoagulation of patients with heparin-induced thrombocytopenia (HIT) may be limited by cross-reaction of HIT antibodies with danaparoid and generation of antibodies during therapy with lepirudin. We used fondaparinux to treat 6 patients with a history of HIT with thromboembolism and 2 patients with thrombocytopenia during low-molecular-weight heparin administration. PMID- 15339692 TI - Effect of pre-competition and altitude training on blood models used to detect erythropoietin abuse by athletes. AB - This study reports blood model scores used for detection of recombinant human erythropoietin (rHuEPO) abuse by athletes. Elite female rowers were monitored prior to their World Championships, including a period spent training at moderate altitude. In contrast to previous results, no substantial increase in model scores was found following altitude exposure. PMID- 15339693 TI - Further characterisation of Hb Okazaki [b93 (F8) Cys-Arg], a rare hemoglobin variant found in a family from Naples, Italy. PMID- 15339694 TI - Soft tissue and skeletal involvement in FIP1L1-PDGFR-alpha positive chronic eosinophilic leukemia: imatinib mesylate may induce complete molecular and imaging remission. PMID- 15339695 TI - Two clonal occurrences of tetrasomy 21 in an atypical chronic myeloid leukemia with wild-type RUNX1 alleles. Additional support for a gene dosage effect of chromosome 21 or RUNX1 in leukemia. AB - Atypical chronic myeloid leukemia (aCML) is a rare leukemic disorder with no specific genetic lesion. Here we demonstrate clonal occurrences of tetrasomy for the long arm of chromosome 21 in a patient with aCML, and a thorough review of the literature provides evidence that this chromosomal anomaly is a so far not recognised recurrent finding in aCML. Further, the timely association of the occurrence of the tetrasomy 21q with acceleration of the leukemia suggests a role for chromosome 21 in leukemic disease progression. The chromosome 21 gene most strongly implicated in both normal and abnormal hematopoiesis is RUNX1. Also, RUNX1 haploinsufficiency due to RUNX1 point mutations characterises the familial platelet disorder with propensity to develop leukemia, and thromboytopenia was a leading feature in the present case. Therefore, an extensive molecular analysis of RUNX1 was performed. However, these analyses did not reveal a mutation, and the results support a gene dosage effect for RUNX1 in myeloid disease similar to observations in lymphoid disease. Patients with aCML and a tetrasomy 21 may form a karyotypically and phenotypically defined subgroup of aCML. PMID- 15339696 TI - T-cell lymphoblastic leukemia/lymphoma in Nijmegen breakage syndrome. AB - Nijmegen breakage syndrome (NBS) is a rare autosomal-recessive disorder characterized by microcephaly, immunodeficiency and predisposition to cancer, mainly B-cell lymphomas. Our 10-years-old female patient with NBS developed T cell lymphoblastic leukemia/lymphoma (TLBL/ALL). The use of standard chemotherapy in our patient, except for cranial irradiation, led to complete and sustained remission of TLBL/ALL. In patients with chromosomal instability syndromes chemotherapy must be modified and radiotherapy must be omitted because of potentially serious toxic complications. Careful prevention of infections, including the use of intravenous immunoglobulin is also essential for successful treatment of lymphoid malignancies in NBS. Immunodeficiency in NBS is profound affecting both humoral and cellular immune system. During long-term follow-up after treatment of malignancy our patient remained free of major infections However, the rise of oligoclonal serum IgM was detected recently. Monitoring of serum IgM concentration may be a useful indicator for early detection of lymphomas in NBS. PMID- 15339697 TI - An inv(16)(p13q22) positive acute myeloid leukaemia relapsing as acute precursor B-cell lymphoblastic leukaemia. AB - We describe a case of a 38-year old male with inv(16)(p13q22) positive acute myeloid leukaemia (AML) with eosinophilia, relapsing after a molecular remission of almost three years. Remarkably, the leukaemia at relapse was identified as a precursor-B-cell acute lymphoblastic leukaemia (B-ALL) by cytology and immunophenotyping, but was inv(16)(p13q22) positive as revealed by interphase FISH, FICTION analysis, and real-time quantitative PCR. Analysis of immunoglobulin and T-cell receptor genes showed a bi-allelic DH2-JH rearrangement at relapse, but not at diagnosis. These findings indicate a myeloid to lymphoid lineage switch from an inv(16)(p13q22) positive leukaemia and show that IGH gene rearrangements can occur in the presence of CBFB-MYH11 fusion transcripts. PMID- 15339698 TI - Type I CD36 deficiency in hematologic disorder. PMID- 15339699 TI - IgA myeloma with quadrangular crystalline inclusions. PMID- 15339700 TI - Epitaph for erythroleukemia. PMID- 15339701 TI - Re: Lange T et al. Quantitative reverse transcription polymerase chain reaction should not replace conventional cytogenetics for monitoring patients with chronic myeloid leukemia during early phase of imatinib therapy. Leukemia 2004; 89: 49 57. PMID- 15339702 TI - Acquired angioedema: a new target for rituximab? PMID- 15339710 TI - A profile of OxyContin addiction. AB - OxyContin is a controlled-released form of oxycodone indicated for the management of moderate to severe pain. OxyContin diversion and abuse has become a major problem in certain areas of the United States, particularly rural areas and Appalachia. This study undertakes an 18-month retrospective chart review at a private freestanding psychiatric facility to develop a profile of OxyContin addicts seeking treatment. There were 579 admissions to the Addictive Disease Unit of this facility from October 2000 to March 2002, with 298 of these admissions being for the treatment of opioid abuse or dependence. One hundred and eighty seven of these individuals were dependent on OxyContin, using an average dose of 184 milligrams of OxyContin per day. The OxyContin dependent individuals tended to show a progression from p.o. use to either snorting or i.v. use. The author concludes that a sociocultural understanding is needed to better treat these addicts as is improved communication between the pain treatment community and the addiction treatment community. PMID- 15339711 TI - Naltrexone and pharmacy benefit management. AB - Naltrexone was approved by the FDA in 1994 for the treatment of alcohol dependence. Despite the potential to make treatment more effective and accessible, naltrexone use remains low by almost any measure. While many of the factors responsible for the slow pace of diffusion are unique to naltrexone and to the organization and delivery of alcohol treatment services, other factors are the same as those that affect the use of prescription medications more generally. Access to third-party coverage and formulary inclusion are necessary conditions for the adoption and diffusion of any new pharmaceutical technology. This paper describes current issues in drug benefit design and formulary coverage decisions, reviews publicly available information on naltrexone coverage by large health insurance programs and pharmaceutical benefit management companies, and examines whether drug benefit design constitutes a barrier to naltrexone use. Our review suggests that naltrexone is widely covered on public and private health plan formularies, though restrictions on use (i.e., quantity limits, prior authorization) are common. PMID- 15339712 TI - Prevalence and characteristics of adolescents patients with co-occurring ADHD and substance dependence. AB - Estimates of co-morbidity of SUD and ADHD in addiction treatment settings range from 30% to 50%. The Schedule II psychostimulant medications, methylphenidate and dextroamphetamine, generally considered to be safe and effective in treating ADHD in adolescent patients, may be risky for an SUD population since individuals with SUD may have a higher likelihood of abusing or diverting the medications. One hundred sixty-two adolescent patients admitted to a residential addictions treatment program were administered a structured interview concerning ADHD and psychostimulant abuse as part of the clinical psychological evaluation administered by the staff psychologist. Results indicate 31% of patients have current ADHD diagnosis and 20% reported illicit diversion of Schedule II medication. One-third of entire adolescent patient population reported prior psychostimulant abuse. Results are discussed in terms of appropriate treatment for adolescents with co-occurring substance abuse or dependence and ADHD. PMID- 15339713 TI - Indentifying co-occurring disorders in adolescent population. AB - A structured diagnostic interview (Practical Adolescent Diagnostic Interview) designed to gather basic information about substance use disorders, other mental health conditions, and related experiences was used in a variety of clinical settings. Anonymous data from 279 adolescents interviewed as part of routine clinical assessments in a variety of clinical programs were analyzed to assess the ability of the questions to identify potential problem areas and to provide a preliminary exploration of interrelationships between those problems. Results demonstrated that the vast majority of individuals manifested indications of multiple problems. For a given diagnostic condition, the trend is for those meeting at least the minimal DSM-IV criteria to exhibit substantially more than the minimum number of symptoms. Internal consistencies for item groups defining the various conditions range from more than .700 to over .900 indicating adequate to excellent internal consistency and reliability. Utility of the instrument for routine clinical use is also discussed. PMID- 15339714 TI - Depression, negative self-image, and suicidal attempts as effects of substance use and substance dependence. AB - The main purpose of this study was to compare, separately by gender, the degree to which cocaine use/abuse, marijuana use/ abuse, and alcohol use/abuse each predict to developing either: a state of depression, or a negative self-image, or a negative personal outlook or for making a suicidal attempt. The study sample of 431 inner-city, African-American subjects, derived from the National Collaborative Perinatal Project (NCPP), were retrieved for the fourth time, at average age 37. RESULTS: (1) For the female sample, there were more predictions and relatively stronger predictions from the use/abuse of and from the dependence on illicit drugs, to a subsequent negative self-image and to a negative personal outlook. For the male sample, there were relatively more predictions to making suicide attempts. (2) Of the three types of substances, cocaine use/abuse predicted to the greatest number of negative outcomes; and alcohol use/abuse predicted to no negative outcomes. PMID- 15339715 TI - Thirst for living of alcoholic patients: medical care's experience. AB - The medical care lavished on terminal and on alcoholic patients leads us to observe an analogy between these two life paths. This raises a question: do links exist between these two conditions? We will present Mr. B's record with alcohol. Analysing alcohol withdrawal allows us to show that during withdrawal, after having experienced similar stages to those described by Kubler-Ross, the alcoholic patient is in a grieving process. When abstinent, Mr. B faces two possibilities: either the grief is incomplete and the drive towards alcohol leads him to die to life; or he remains abstinent, with the help sometimes of what we call 'support dyads'. All this presupposes that the initial stages of dying to alcohol, followed by a psychic work of dis-fusion, have been brought to a successful conclusion. Links do exist between these two life paths, in the sense that these stages enable them to face the major trauma of their lives, e.g. alcoholism and serious illness. PMID- 15339716 TI - A psychometric analysis of the dimension rating system--2nd edition. AB - Results from a two-year beta test, in a strong version of a Minnesota Model program, showed differences (p < .001) in all six ASAM dimensions rated for level II.1 and III.7 admissions by the DRS-2. Inter-rater reliability was .913, and intra-class correlations in year one admissions to level III.7 (r = .576 and .720, n = 578) and level II.1 (r = .462 and .632, n = 323) were comparable to internal consistency reported in year two. Inter-class correlations for both admission and discharge ratings occurred as predicted by the measurement design, and little evidence of rater drift for mean dimension scores or rater reliability was found. It was concluded that the DRS-2 is a reliable and valid method for rating impediments to treatment progress that is consistent with the structure and intent of the PPC-2. Limitations to the generalizability of these findings and ramifications for the PPC-2R are discussed. PMID- 15339717 TI - Consumption of caroltenoid-rich foods and central vision loss:a matched case controlled study in Kansas. AB - The purpose of this study was to explore the association between consumption of carotenoid-rich foods and risk of central vision loss in 168 rural elders. Food consumption was estimated using a food frequency questionnaire. Median servings per week were used as the cut points for consumption levels. Odds ratios (OR) and 95% confidence intervals (CI) were calculated by using discordant pairs. High consumption of corn bread indicated significant association with central vision loss (OR 0.4; 95% CI 0.2, 0.9). Insignificant inverse associations were observed for seven other foods and the summary index of all selected foods. Prospective studies are needed to further evaluate the potential protective influence of carotenoid-rich foods on central vision loss among older adults. PMID- 15339718 TI - Nutrition risk screening: the interrelationship of food insecurity, food intake, and unintentional weight change among homebound elders. AB - Nutrition risk screening is a key component of the homedelivered meals program. To examine direct and indirect relationships among individual components of nutrition risk, path analysis was conducted on routinely collected data from 908 homebound elders who received home-delivered meals. The good fit of the model revealed that specific nutrition risk factors and indicators of nutritional risk were directly and indirectly associated with meal frequency and unintended weight change. With the heightened vulnerability for poor nutritional health among homebound elders who report food insecurity, policymakers and service providers should strengthen efforts to target individual components of nutrition risk rather than aggregate scores or categorical measures. PMID- 15339719 TI - Food label use by older Americans: data from the Continuing Survey of Food Intakes by Individuals and the Diet and Health Knowledge Survey 1994-96. AB - This study examined older Americans' use of food labels as a tool to moderate dietary risk factors for heart disease. Data from the USDA's Continuing Survey of Food Intakes by Individual (CSFII) 1994- 96 and the Diet and Health Knowledge Survey (DHKS) from 2,846 respondents aged 51 years and older was used. Indices were constructed to measure and correlate dietary intake, label use and understanding, and health status data. Understanding and use of food labels was lower among older age groups, while heart-related health problems increased. Use of food labels and percent energy intake from fat were inversely related (P < 0.01). Older Americans would benefit from education on food label use and interpretation to decrease their dietary risk factors for heart disease. PMID- 15339720 TI - Nutritional risk and body composition in free-living elderly participating in congregate meal-site programs. AB - Early detection of nutritional risk in free-living elderly is critical in healthcare, yet comprehensive measurements are time consuming and can be frustrating to both health professionals and elderly. In addition, body composition measurements provide information regarding fat and fat-free mass that have been linked to morbidity and mortality in elderly. In this study, nutritional risk was assessed in 69 elderly, aged 50-90 years, attending congregate meal-site programs, using Mini Nutritional Assessment, and body composition was assessed by bioelectric impedance. Analysis revealed that 31.9% of the elderly were at risk for malnutrition and 2.9% were malnourished. Males had significantly greater body weight, height and fat-free mass, and females had significantly greater body fat as percentage of body weight, but there was no gender difference in nutritional risk. Of elderly, 36.2% had body mass index > 85th percentile and 8.7% < 15th percentile using national population reference standards. Age-related decline in fat-free mass was an early indicator of changes in body composition. PMID- 15339722 TI - Men can cook! Development, implementation, and evaluation of a senior men's cooking group. AB - This study reports on the process and outcome evaluation of a community-based nutrition and cooking education program for senior men. As part of Evergreen Action Nutrition, a community-organized, nutrition education program, a registered dietitian led a Men's Cooking Group in a seniors' recreation facility. Written questionnaires were completed by most of the men (n = 19) at the beginning and end of the evaluation year, and ten men participated in personal key informant interviews. The majority of participants gained cooking confidence, increased their cooking activities at home, developed healthy cooking skills, and improved cooking variety through the program. The men also identified social benefits to the program. Overall, this preliminary evaluation suggests that community-based nutrition and cooking education for older men is a beneficial nutrition education activity. PMID- 15339723 TI - Reaching out to racial/ethnic minority older persons for elderly nutrition programs. AB - This paper reports findings from a study that was conducted to identify perceived barriers to racial/ethnic minority elders' participation in an elderly nutrition program (ENP) in a large metropolitan area and effective strategies for reaching out to them. The data were collected from a survey with the ENP's staff and volunteers and three focus group discussions with professionals working with minority elders and minority community leaders. The study participants identified as their perceived barriers: the lack of information or misinformation; culturally driven reluctance to ask for outside help; fear and distrust of formal systems; lack of ethnic menus in the program; discomfort due to cultural differences; and inaccessibility and inadequacy of transportation. Recommended outreach strategies included: involvement of family members in the information dissemination process; establishment of good working relationships with community leaders and contact with key older persons; diversification of menus and increased use of food enhancements; increase in cultural activities/programs in congregate dining centers; solicitation of input from current participants; provision of intergenerational programs; recruitment of volunteer drivers from the minority community; location of the program in ethnic enclaves or places where minority elders can easily congregate; and improvement in transportation services. PMID- 15339728 TI - Laser ablation-combustion-GC-IRMS--a new method for online analysis of intra annual variation of delta13C in tree rings. AB - We present a new, rapid method for high-resolution online determination of delta13C in tree rings, combining laser ablation (LA), combustion (C), gas chromatography (GC) and isotope ratio mass spectrometry (IRMS) (LA-C-GC-IRMS). Sample material was extracted every 6 min with a UV-laser from a tree core, leaving 40-microm-wide holes. Ablated wood dust was combusted to CO2 at 700 degrees C, separated from other gases on a GC column and injected into an isotope ratio mass spectrometer after removal of water vapor. The measurements were calibrated against an internal and an external standard. The tree core remained intact and could be used for subsequent dendrochronological and dendrochemical analyses. Cores from two Scots pine trees (Pinus sylvestris spp. sibirica Lebed.) from central Siberia were sampled. Inter- and intra-annual patterns of delta13C in whole-wood and lignin-extracted cores were indistinguishable apart from a constant offset, suggesting that lignin extraction is unnecessary for our method. Comparison with the conventional method (microtome slicing, elemental analysis and IRMS) indicated high accuracy of the LA-C-GC-IRMS measurements. Patterns of delta13C along three parallel ablation lines on the same core showed high congruence. A conservative estimate of the precision was +/- 0.24 per thousand. Isotopic patterns of the two Scots pine trees were broadly similar, indicating a signal related to the forest stand's climate history. The maximum variation in delta13C over 22 years was about 5 per thousand, ranging from -27 to -22.3 per thousand. The most obvious pattern was a sharp decline in delta13C during latewood formation and a rapid increase with spring early growth. We conclude that the LA-C-GC-IRMS method will be useful in elucidating short-term climate effects on the delta13C signal in tree rings. PMID- 15339729 TI - Growth and transpiration of Japanese cedar (Cryptomeria japonica) and Hinoki cypress (Chamaecyparis obtusa) seedlings in response to soil water content. AB - To investigate the effects of soil water content on growth and transpiration of Japanese cedar (Cryptomeria japonica D. Don) and Hinoki cypress (Chamaecyparis obtusa (Siebold et Zucc.) Endl.), potted seedlings were grown in well-watered soil (wet treatment) or in drying soil (dry treatment) for 12 weeks. Seedlings in the wet treatment were watered once every 2 or 3 days, whereas seedlings in the dry treatment were watered when soil water content (Theta; m3 m(-3)) reached 0.30, equivalent to a soil matric potential of -0.06 MPa. From Weeks 7 to 12 after the onset of the treatments, seedling transpiration was measured by weighing the potted seedlings. After the last watering, changes in transpiration rate during soil drying were monitored intensely. The dry treatment restricted aboveground growth but increased biomass allocation to the roots in both species, resulting in no significant treatment difference in whole-plant biomass production. The species showed similar responses in relative growth rate (RGR), net assimilation rate (NAR) and shoot mass ratio (SMR) to the dry treatment. Although NAR did not change significantly in either C. japonica or C. obtusa as the soil dried, the two species responded differently to the dry treatment in terms of mean transpiration rate (E) and water-use efficiency (WUE), which are parameters that relate to NAR. In the dry treatment, both E and WUE of C. japonica were stable, whereas in C. obtusa, E decreased and WUE increased (E and WUE counterbalanced to maintain a constant NAR). Transpiration rates were lower in C. obtusa seedlings than in C. japonica seedlings, even in well-watered conditions. During soil drying, the transpiration rate decreased after Theta reached about 0.38 (-0.003 MPa) in C. obtusa and 0.32 (-0.028 MPa) in C. japonica. We conclude that C. obtusa has more water-saving characteristics than C. japonica, particularly when water supply is limited. PMID- 15339730 TI - Integrating within-crown variation in net photosynthesis in loblolly and slash pine families. AB - We examined photosynthetic characteristics of two fast- and two slow-growing half sib families of both loblolly pine (Pinus taeda L.) and slash pine (Pinus elliottii var. elliottii Engelm.) on two sites in northern Florida to: (1) quantify variation in light-saturated net photosynthesis (Amax) associated with vertical crown position and foliage age; (2) quantify the amount and distribution of leaf area by foliage age class; and (3) determine whether photosynthetic indices, ranging from leaf-level through whole-crown Amax, were related to growth differences among species and families. In both species, leaf-level Amax was higher in more recently formed foliage both within the same year (where Amax in the third flush averaged 10 to 30% higher than Amax in the first flush) and between years (where Amax in current-year foliage averaged 20 to 40% higher than Amax in 1-year-old foliage). When expressed on a leaf area basis, Amax of current year foliage was higher in slash pine than in loblolly pine, but Amax expressed on a mass basis did not differ between species. Loblolly pine had higher whole tree leaf area than slash pine, whereas whole-tree Amax did not differ between species. When the mean values for fast-growing families were compared with the mean values for slow-growing families, there were no differences in leaf-level characteristics, whereas at the whole-tree level, fast-growing families had higher leaf area and whole-tree Amax than slow-growing families in both species. When comparisons were made among the individual fast- and slow-growing families, however, results were more variable. In both species, stem volume growth was strongly correlated with whole-tree Amax, with most of the strength of the correlation deriving from the relationship between volume growth and tree leaf area. PMID- 15339731 TI - Changes in polyamine concentration associated with aging in Pinus radiata and Prunus persica. AB - Endogenous polyamine concentrations in apical buds of peach (Prunus persica (L.) Batsch cv. 'Hakuho') and monterey pine (Pinus radiata D. Don) seedlings, saplings and mature trees were analyzed by high performance liquid chromatography coupled with fluorescence detection to establish physiological correspondences and disparities between the species. Although there were many differences in polyamine metabolism between peach and monterey pine, several similarities were found. Specifically, in both species, the ratio between concentrations of free polyamines and low molecular-weight polyamine conjugates was greater than and less than unity in juvenile and adult plants, respectively. Free putrescine concentration decreased with age in both species. PMID- 15339732 TI - Silver birch and climate change: variable growth and carbon allocation responses to elevated concentrations of carbon dioxide and ozone. AB - We studied the effects of elevated concentrations of carbon dioxide ([CO2]) and ozone ([O3]) on growth, biomass allocation and leaf area of field-grown O3 tolerant (Clone 4) and O3-sensitive clones (Clone 80) of European silver birch (Betula pendula Roth) trees during 1999-2001. Seven-year-old trees of Clones 4 and 80 growing outside in open-top chambers were exposed for 3 years to the following treatments: outside control (OC); chamber control (CC); 2 x ambient [CO2] (EC); 2 x ambient [O3] (EO); and 2 x ambient [CO2] + 2 x ambient [O3] (EC+EO). When the results for the two clones were analyzed together, elevated [CO2] increased tree growth and biomass, but had no effect on biomass allocation. Total leaf area increased and leaf abscission was delayed in response to elevated [CO2]. Elevated [O3] decreased dry mass of roots and branches and mean leaf size and induced earlier leaf abscission in the autumn; otherwise, the effects of elevated [O3] were small across the clones. However, there were significant interactions between elevated [CO2] and elevated [O3]. When results for the clones were analyzed separately, stem diameter, volume growth and total biomass of Clone 80 were increased by elevated [CO2] and the stimulatory effects of elevated [CO2] on stem volume growth and total leaf area increased during the 3 year study. Clone 80 was unaffected by elevated [O3]. In Clone 4, elevated [O3] decreased root and branch biomass by 38 and 29%, respectively, whereas this clone showed few responses to elevated [CO2]. Elevated [CO2] significantly increased total leaf area in Clone 80 only, which may partly explain the smaller growth responses to elevated [CO2] of Clone 4 compared with Clone 80. Although we observed responses to elevated [O3], the responses to the EC+EO and EC treatments were similar, indicating that the trees only responded to elevated [O3] under ambient [CO2] conditions, perhaps reflecting a greater quantity of carbohydrates available for detoxification and repair in elevated [CO2]. PMID- 15339733 TI - Gas in stems: abundance and potential consequences for tree biomechanics. AB - Secondary xylem of woody plants has a large volumetric proportion of gas occupying spaces that would otherwise be filled with water. We examined whether these gas-filled voids have a mechanical role by either decreasing the fresh mass the tree must support (by replacing some of the water with gas) or by providing inexpensive filler to increase stem diameter (thereby increasing the second moment of area at the expense of the modulus of elasticity and modulus of rupture). Calculations from published data show that temperate softwood species (n = 26) average 18 and 50% gas by volume for sapwood and heartwood, respectively; temperate hardwood species (n = 31) average 26% gas by volume in both the sapwood and heartwood; and tropical species (n = 52) with mixed sapwood and heartwood have 18% gas by volume. In this paper, we develop equations to show how gas affects the mechanical behavior of tree stems, and describe model results to show how gas affects mechanical stability, based on mass and stem diameters for six 34-year-old Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees. For the same applied load, modeled stems in which the gas space was filled with water differed in their surface stresses by < 2% from modeled stems in the native state (partially gas-filled), indicating no practical benefit from a reduction in stem mass due to gas. A second modeling scenario compared the native state to stems in which gas was removed and stem diameters decreased (and material properties adjusted to concur with the increased wood density) to conserve mass. Removal of the gas-filled voids resulted in up to 41% higher surface stress for the same applied load, caused by a decrease in the second moment of area greater than the increase in modulus of elasticity. Trees with gas removed had higher modulus of rupture, but could withstand up to 14% lower maximum wind forces than trees in their native state, suggesting a biomechanical role for the gas if the model assumptions are valid. The gas content may, however, have evolved in response to pressures unrelated to biomechanics. We discuss some of its potential effects on sapwood physiology. PMID- 15339734 TI - Changes in water status and proline and abscisic acid concentrations in developing somatic embryos of pedunculate oak (Quercus robur) during maturation and germination. AB - Somatic embryos of oak (Quercus robur L.) were matured on P24 media differing in gel strength (0.8, 0.9 and 1.0% (w/v) agar). Viscosity and osmotic potential (Psipi,medium) of the media were determined. Developing cotyledonary embryos were analyzed at maturity Stages I-III for water content, osmotic potential (Psipi,embryo) and concentrations of abscisic acid (ABA) and proline. Proliferation of embryogenic tissue, germination rates and the number of embryos formed were also determined in order to relate embryo quality to physiological parameters. Viscosity increased with agar concentration, a phenomenon apparently related to water availability. Many Stage III embryos with high germination potentials were obtained on P24 medium containing 1.0% agar. Embryo water content decreased progressively from 94 to 80% during embryo maturation. Stage I and II embryos that matured on media containing 0.8 or 0.9% agar had similar values of Psipi,embryo, whereas Psipi,embryo of Stage III embryos that matured on medium containing 1.0% agar was significantly lower, although Psipi,medium was unaffected by gel strength. Stage III embryos showed a nearly 16-fold increase in proline concentration and a 50% decrease in ABA concentration compared with Stage I embryos. We conclude that tissue water status and a complex relationship between ABA and proline concentrations, modulated by medium gel strength, are important factors in the maturation process and the quality of oak somatic embryos. PMID- 15339735 TI - Effect of parent genotype on somatic embryogenesis in Scots pine (Pinus sylvestris). AB - Controlled crosses of seven Scots pine (Pinus sylvestris L.) trees produced 49 families that included both reciprocals and selfings. Embryogenic cultures were initiated from immature megagametophytes and after 6 months in maintenance culture, mature somatic embryos were produced from the surviving 166 lines. The effect of parent genotypes on the cultures was evaluated at initiation of the tissue culture period, after 6 months in maintenance culture and at embryo maturation. The effect of the maternal parent was most pronounced at culture initiation. After 6 months in tissue culture, the maternal effect had decreased and the effects of both parents were significant. By the somatic embryo maturation stage, the maternal effect was still considerable but the paternal effect was no longer detectable. There was little correlation between the ranking of mothers and fathers, indicating that the maternal effect was caused by factors other than the paternal effect. No mother x father interaction was found, indicating that mothers successful at initiation and after 6 months in tissue culture, pollinated by any of the successful fathers, produced somatic lines and mature somatic embryos. PMID- 15339736 TI - Responses of eucalypt species to aluminum: the possible involvement of low molecular weight organic acids in the Al tolerance mechanism. AB - Aluminum (Al) tolerance mechanisms in crop plants have been extensively researched, but our understanding of the physiological mechanisms underlying Al tolerance in trees is still limited. To investigate Al tolerance in eucalypts, seedlings of six species (Eucalyptus globulus Labill., Eucalyptus urophylla S.T. Blake, Eucalyptus dunnii Maiden, Eucalyptus saligna Sm., Eucalyptus cloeziana F. J. Muell. and Eucalyptus grandis w. Hill ex Maiden) and seedlings of six clones of Eucalyptus species were grown for 10 days in nutrient solutions containing Al concentrations varying from 0 to 2.5 microM (0 to 648 microM Al3+ activities). Root elongation of most species was inhibited only by high Al3+ activities. Low to intermediate Al3+ activities were beneficial to root elongation of all species and clones. Among the species tested, E. globulus and E. urophylla were more tolerant to Al toxicity, whereas E. grandis and E. cloeziana were more susceptible to Al-induced damage. Although E. globulus seedlings were tolerant to Al toxicity, they were highly sensitive to lanthanum (La), indicating that the tolerance mechanism is specific for Al. Fine roots accumulated more Al and their elongation was inhibited more than that of thick roots. In E. globulus, accumulation of Al in root tips increased linearly with increasing Al concentration in the nutrient solution. The majority of Al taken up was retained in the root system, and the small amounts of Al translocated to the shoot system were found mainly in older leaves. No more than 60% of the Al in the thick root tip was in an exchangeable form in the apoplast that could be removed by sequential citrate rinses. Gas chromatography/mass spectrometry and ion chromatography analyses indicated that root exposure to Al led to a greater than 200% increase in malic acid concentration in the root tips of all eucalypt species. The increase in malate concentration in response to Al treatment correlated with the degree of Al tolerance of the species. A small increase in citric acid concentration was also observed in all species, but there were no consistent changes in the concentrations of other organic acids in response to Al treatment. In all eucalypt species, Al treatment induced the secretion of citric and malic acid in root exudates, but no trend with respect to Al tolerance was observed. Thus, although malate and citrate exudation by roots may partially account for the overall high Al tolerance of these eucalypt species, it appears that tolerance is mainly derived from the internal detoxification of Al by complexation with malic acid. PMID- 15339737 TI - Induction of callose in roots of Norway spruce seedlings after short-term exposure to aluminum. AB - Callose (1,3-beta-glucan) is a suggested physiological indicator of aluminum (Al) toxicity in crop plants. It is not known if callose serves a similar function in forest trees, because quantitative data on callose formation in tree roots are limited, particularly under controlled conditions. To evaluate callose as a physiological indicator of Al toxicity in tree roots, we quantified callose formation in roots of Norway spruce (Picea abies (L.) Karst.) seedlings. Seedlings were grown in simulated soil solutions in the presence or absence (control) of Al under controlled conditions. In seedlings grown in solutions containing 280 microM Al, callose concentrations in roots were twice as high as control values after 6 h of Al treatment and 5 times higher than control values after 1 day. Thereafter, root callose concentrations gradually decreased and were only twice as high as control values after 7 days. The presence of various Al concentrations in the simulated soil solutions indicated that callose was induced by a relatively low Al concentration (84 microM). We conclude that callose in tree roots is an indicator of Al toxicity. PMID- 15339738 TI - Radial profiles of sap flow with increasing tree size in maritime pine. AB - We investigated the radial variation of sap flow within sapwood below the live crown in relation to tree size in 10-, 32-, 54- and 91-year-old maritime pine stands (Pinus pinaster Ait.). Radial variations were determined with two thermal dissipation sensors; one measured sap flux in the outer 20 mm of the xylem (Jref), whereas the other was moved radially across the sapwood in 20-mm increments to measure sap flux at multiple depths (Jref). For all tree sizes, sap flow ratios (Ri = JiJref (-1)) declined with increasing sapwood depth, but the decrease was steeper in trees with large diameters. Correction factors (C) were calculated to extrapolate Jref for an estimate of whole-tree sap flux. A negative linear relationship was established between stem diameter and C, the latter ranging from 0.6 to 1.0. We found that neglecting these radial corrections in 10 , 32-, 54- and 91-year-old trees would lead to overestimation of stand transpiration by 4, 14, 26 and 47%, respectively. Therefore, it is necessary to account for the differential radial profiles of sap flow in relation to tree size when comparing tree transpiration and hydraulic properties among trees differing in size. PMID- 15339739 TI - Wood density and anatomy of water-limited eucalypts. AB - We hypothesized that seedlings grown under water-limited conditions would develop denser wood than seedlings grown under well-watered conditions. Three Eucalyptus species (E. grandis Hill (ex Maiden), E. sideroxylon Cunn. (ex Woolls) and E. occidentalis Endl.) were grown in a temperature-controlled greenhouse for 19 weeks with watering treatments (well-watered and water-limited) applied at six weeks. The water-limitation treatment consisted of four drought cycles. Wood density increased by between 4 and 13% in the water-limited seedlings, but this increase was mainly due to extractive compounds embedded in the cell wall matrix. Once these compounds were removed, the increase was 0-9% and was significant for E. grandis only. Water-limitation significantly reduced mean vessel lumen area; however, this was balanced by a trend toward greater vessel frequency in water limited plants, and consequently there was no difference in the proportion of stem area allocated to vessels. Conduit efficiency value was lowest in the water limited plants, indicating that there was a cost in terms of stem hydraulic conductivity for decreasing vessel lumen area. Wood density was negatively correlated with vessel lumen fraction in well-watered plants, but this relationship broke down in the water-limited plants, possibly because of the significantly larger proportion of the stem taken up by pith in water-limited seedlings. Diurnal variation in leaf water potential was positively correlated with wood density in well-watered plants. This relationship did not hold in the water-limited plants owing to the collapse of the pressure gradient between soil and leaf. We conclude that drought periods of greater than 1 month are required to increase wood density in these species and that increases in wood density appear to result in diminished capacity to supply water to leaves. PMID- 15339740 TI - Drought-induced changes in flavonoids and other low molecular weight antioxidants in Cistus clusii grown under Mediterranean field conditions. AB - Mediterranean plants have evolved a complex antioxidant defense system to cope with summer drought. Flavonoids, and particularly flavanols and flavonols, are potent in vitro antioxidants, but their in vivo significance within the complex network of antioxidant defenses remains unclear, especially in plant responses to stress. To gain insight into the role of flavonoids in the antioxidant defense system of Cistus clusii Dunal, we evaluated drought-induced changes in flavonoids in leaves and compared the response of these compounds with that of other low molecular weight antioxidants (ascorbic acid, tocopherols and carotenoids). Among the antioxidant flavonoids analyzed, epigallocatechin gallate was present in the greatest concentrations (up to about 5 micromol dm(-2)). Other flavanols, such as epicatechin and epicatechin gallate, were found at concentrations below 0.25 and 0.03 micromol dm(-2), respectively. Neither of the antioxidant flavonols analyzed, quercetin and kaempferol, were detected in C. clusii leaves. Epigallocatechin gallate, ascorbic acid and alpha-tocopherol concentrations increased to a similar extent (up to 2.8-, 2.6- and 3.3-fold, respectively) in response to drought, but the kinetics of the drought-induced increases differed. Epigallocatechin gallate, epicatechin and epicatechin gallate concentrations increased progressively during drought, reaching maximum values after 30 days of stress. Ascorbic acid concentrations increased twofold after 15 days of drought, and maximum values were attained after 50 days of drought. In contrast, alpha tocopherol concentrations remained constant during the first 30 days of drought, but increased sharply by 3.3-fold after 50 days of drought. The maximum efficiency of photosystem II photochemistry and the extent of lipid peroxidation remained constant throughout the drought period, whereas the redox state of ascorbic acid and alpha-tocopherol shifted toward their reduced forms in response to drought, indicating that the concerted action of low molecular weight antioxidants may help prevent oxidative damage in plants. PMID- 15339741 TI - Mitochondrial metabolism reveals a functional architecture in intact islets of Langerhans from normal and diabetic Psammomys obesus. AB - The cells within the intact islet of Langerhans function as a metabolic syncytium, secreting insulin in a coordinated and oscillatory manner in response to external fuel. With increased glucose, the oscillatory amplitude is enhanced, leading to the hypothesis that cells within the islet are secreting with greater synchronization. Consequently, non-insulin-dependent diabetes mellitus (NIDDM; type 2 diabetes)-induced irregularities in insulin secretion oscillations may be attributed to decreased intercellular coordination. The purpose of the present study was to determine whether the degree of metabolic coordination within the intact islet was enhanced by increased glucose and compromised by NIDDM. Experiments were performed with isolated islets from normal and diabetic Psammomys obesus. Using confocal microscopy and the mitochondrial potentiometric dye rhodamine 123, we measured mitochondrial membrane potential oscillations in individual cells within intact islets. When mitochondrial membrane potential was averaged from all the cells in a single islet, the resultant waveform demonstrated clear sinusoidal oscillations. Cells within islets were heterogeneous in terms of cellular synchronicity (similarity in phase and period), sinusoidal regularity, and frequency of oscillation. Cells within normal islets oscillated with greater synchronicity compared with cells within diabetic islets. The range of oscillatory frequencies was unchanged by glucose or diabetes. Cells within diabetic (but not normal) islets increased oscillatory regularity in response to glucose. These data support the hypothesis that glucose enhances metabolic coupling in normal islets and that the dampening of oscillatory insulin secretion in NIDDM may result from disrupted metabolic coupling. PMID- 15339742 TI - Magnetic resonance spectroscopy to measure hepatic triglyceride content: prevalence of hepatic steatosis in the general population. AB - Despite the increasing prevalence of nonalcoholic fatty liver disease (NAFLD), the criteria used to diagnose the disorder remain poorly defined. Localized proton magnetic resonance spectroscopy (MRS) accurately measures hepatic triglyceride content (HTGC) but has been used only in small research studies. Here, MRS was used to analyze the distribution of HTGC in 2,349 participants from the Dallas Heart Study (DHS). The reproducibility of the procedure was validated by showing that duplicate HTGC measurements were high correlated (r = 0.99, P < 0.001) and that the coefficient of variation between measurements was low (8.5%). Intake of a high-fat meal did not significantly affect the measurements, and values were similar when measurements were made from the right and left hepatic lobes. To determine the "upper limit of normal" for HTGC, the distribution of HTGC was examined in the 345 subjects from the DHS who had no identifiable risk factors for hepatic steatosis (nonobese, nondiabetic subjects with minimal alcohol consumption, normal liver function tests, and no known liver disease). The 95th percentile of HTGC in these subjects was 5.56%, which corresponds to a hepatic triglyceride level of 55.6 mg/g. With this value as a cutoff, the prevalence of hepatic steatosis in Dallas County was estimated to be 33.6%. Thus MRS provides a sensitive, quantitative, noninvasive method to measure HTGC and, when applied to a large urban US population, revealed a strikingly high prevalence of hepatic steatosis. PMID- 15339743 TI - Role of CD38 in myometrial Ca2+ transients: modulation by progesterone. AB - Oxytocin-induced Ca(2+) transients play an important role in myometrial contractions. Here, using a knockout model, we found that the enzyme CD38, responsible for the synthesis of the second messenger cyclic ADP-ribose (cADPR), plays an important role in the oxytocin-induced Ca(2+) transients and contraction. We also observed that CD38 is necessary for TNF-alpha-increased agonist-stimulated Ca(2+) transients in human myometrial cells. We provide experimental evidence that the TNF-alpha effect is mediated by increased expression of the enzyme CD38. First, we observed that TNF-alpha increased oxytocin-induced Ca(2+) transients and CD38 expression in human myometrial cells. Moreover, using small interference RNA technology, we observed that TNF-alpha stimulation of agonist-induced Ca(2+) transients was abolished by blocking the expression of CD38. In control experiments, we observed that activation of the component of the TNF-alpha signaling pathway, NF-kappaB, was not affected by the treatments. Finally, we observed that the effects of TNF-alpha on CD38 cyclase and oxytocin-induced Ca(2+) transients are abolished by progesterone. In conclusion, we provide the first experimental evidence that CD38 is important for myometrial Ca(2+) transients and contraction. Moreover, CD38 is necessary for the TNF-alpha-mediated augmentation of agonist-induced Ca(2+) transients in myometrial cells. We propose that the balance between cytokines and placental steroids regulates the expression of CD38 in vivo and cell responsiveness to oxytocin. PMID- 15339744 TI - Evidence against a role for insulin-signaling proteins PI 3-kinase and Akt in insulin resistance in human skeletal muscle induced by short-term GH infusion. AB - Prolonged growth hormone (GH) excess is known to be associated with insulin resistance, but the underlying mechanisms remain unknown. The aim of this study was to assess the impact of GH on insulin-stimulated glucose metabolism and insulin signaling in human skeletal muscle. In a cross-over design, eight healthy male subjects (age 26.0 +/- 0.8 yr and body mass index 24.1 +/- 0.5 kg/m2) were infused for 360 min with either GH (Norditropin, 45 ng.kg(-1).min(-1)) or saline. During the final 180 min of the infusion, a hyperinsulinemic euglycemic clamp was performed (insulin infusion rate: 1.2 mU.kg(-1).min(-1)). Muscle biopsies from vastus lateralis were taken before GH/saline administration and after 60 min of hyperinsulinemia. GLUT4 content and insulin signaling, as assessed by insulin receptor substrate (IRS)-1-associated phosphatidylinositol 3-kinase and Akt activity were determined. GH levels increased to a mean (+/-SE) level of 20.0 +/- 2.3 vs. 0.5 +/- 0.2 microg/l after saline infusion (P < 0.01). During GH infusion, the glucose infusion rate during hyperinsulinemia was reduced by 38% (P < 0.01). In both conditions, free fatty acids were markedly suppressed during hyperinsulinemia. Despite skeletal muscle insulin resistance, insulin still induced a similar approximately 3-fold rise in IRS-1-associated PI 3-kinase activity (269 +/- 105 and 311 +/- 71% compared with baseline, GH vs. saline). GH infusion did not change Akt protein expression, and insulin caused an approximately 13-fold increase in Akt activity (1,309 +/- 327 and 1,287 +/- 173%) after both GH and saline infusion. No difference in total GLUT4 content was noted (114.7 +/- 7.4 and 107.6 +/- 16.7 arbitrary units, GH vs. saline, compared with baseline). In conclusion, insulin resistance in skeletal muscle induced by short term GH administration is not associated with detectable changes in the upstream insulin-signaling cascade or reduction in total GLUT4. Yet unknown mechanisms in insulin signaling downstream of Akt may be responsible. PMID- 15339745 TI - Mechanism, temporal patterns, and magnitudes of the metabolic responses to the KATP channel agonist diazoxide. AB - To assess the mechanism, temporal patterns, and magnitudes of the metabolic responses to the ATP-dependent potassium channel agonist diazoxide, neuroendocrine and metabolic responses to intravenous diazoxide (saline, 1.0 and 2.0 mg/kg) and oral diazoxide (placebo, 4.0 and 6.0 mg/kg) were assessed in healthy young adults. Intravenous diazoxide produced rapid, but transient, decrements (P = 0.0023) in plasma insulin (e.g., nadirs of 2.8 +/- 0.5 and 1.8 +/ 0.3 microU/ml compared with 7.0 +/- 1.0 microU/ml after saline at 4.0-7.5 min) and C-peptide (P = 0.0228) associated with dose-related increments in plasma glucose (P = 0.0044) and serum nonesterified fatty acids (P < 0.0001). After oral diazoxide, plasma insulin appeared to decline, as did C-peptide, again associated with dose-related increments in plasma glucose (P < 0.0001) and serum nonesterified fatty acids (P = 0.0141). Plasma glucagon, as well as cortisol and growth hormone, was not altered. Plasma epinephrine increased (P = 0.0215) slightly only after intravenous diazoxide. There were dose-related increments in plasma norepinephrine (P = 0.0038 and P = 0.0005, respectively), undoubtedly reflecting a compensatory sympathetic neural response to vasodilation produced by diazoxide, but these would not raise plasma glucose or serum nonesterified fatty acid levels. Thus selective suppression of insulin secretion, without stimulation of glucagon secretion, raised plasma glucose and serum nonesterified fatty acid concentrations. These findings define the temporal patterns and magnitudes of the metabolic responses to diazoxide and underscore the primacy of regulated insulin secretion in the physiological regulation of postabsorptive carbohydrate and lipid metabolism. PMID- 15339746 TI - Time course of the hepatic adaptation to TPN: interaction with glycogen depletion. AB - In response to chronic (5 days) TPN, the liver becomes a major site of glucose disposal, removing approximately 45% (4.5 mg.kg(-1).min(-1)) of exogenous glucose. Moreover, approximately 70% of glucose is not stored but released as lactate. We aimed to determine in chronically catheterized conscious dogs the time course of adaptation to TPN and the glycogen depletion impact on early time course. After an 18-h (n = 5) fast, TPN was infused into the inferior vena cava for 8 (n = 5) or 24 h (n = 6). A third group, of 42-h-fasted animals (n = 6), was infused with TPN for 8 h. TPN was infused at a rate designed to match the dog's calculated basal energy and nitrogen requirements. NHGU (-2.3 +/- 0.1 to 2.2 +/- 0.7 to 3.9 +/- 0.6 vs. -1.7 +/- 0.3 to 1.1 +/- 0.5 to 2.9 +/- 0.4 mg.kg(-1).min( 1), basal to 4 to 8 h, 18 vs. 42 h) and net hepatic lactate release (0.7 +/- 0.3 to 0.6 +/- 0.1 to 1.4 +/- 0.2 vs. -0.6 +/- 0.1 to 0.1 +/- 0.1 to 0.8 +/- 0.1 mg.kg(-1).min(-1), basal to 4 to 8 h) increased progressively. Net hepatic glycogen repletion and tracer determined that glycogen syntheses were similar. After 24 h of TPN, NHGU (5.4 +/- 0.6 mg.kg(-1).min(-1)) and net hepatic lactate release (2.6 +/- 0.4 mg.kg(-1).min(-1)) increased further. In summary, 1) most hepatic adaptation to TPN occurs within 24 h after initiation of TPN, and 2) prior glycogen depletion does not augment hepatic adaptation rate. PMID- 15339747 TI - Leucine reduces the duration of insulin-induced PI 3-kinase activity in rat skeletal muscle. AB - Leucine (Leu) is known to stimulate translation initiation of protein synthesis at mammalian target of rapamycin (mTOR) in the insulin signaling pathway. However, potential feedback from mTOR to upstream aspects of the insulin signaling pathway remains controversial. This study evaluates the impact of a physiological oral dose of Leu and/or carbohydrate (CHO) on upstream elements of the insulin signaling pathway using phosphatidylinositol 3-kinase (PI 3-kinase) activity and glucose uptake as markers for insulin sensitivity and glucose homeostasis. Rats (approximately 200 g) were fasted 12 h and administered oral doses of CHO (1.31 g glucose, 1.31 g sucrose), Leu (270 mg), or CHO plus Leu. Animals were killed at 15, 30, 60, and 90 min after treatment. Plasma and gastrocnemius muscles were collected for analyses. Treatments were designed to produce elevated blood glucose and insulin with basal levels of Leu (CHO); elevated Leu with basal levels of glucose and insulin (Leu); or a combined increase of glucose, insulin, and Leu (CHO + Leu). The CHO treatment stimulated PI 3-kinase activity and glucose uptake with no effect on the downstream translation initiation factor eIF4E. Leu alone stimulated the release of the translation initiation factor eIF4E from 4E-BP1 with no effects on PI 3-kinase activity or glucose uptake. The CHO + Leu treatment reduced the magnitude and duration of the PI 3-kinase response but maintained glucose uptake similar to the CHO treatment and eIF4E levels similar to the Leu treatment. These findings demonstrate that Leu reduces insulin-stimulated PI 3-kinase activity while increasing downstream translation initiation and with no effect on net glucose transport in skeletal muscle. PMID- 15339748 TI - Respiratory uncoupling by UCP1 and UCP2 and superoxide generation in endothelial cell mitochondria. AB - Mitochondria represent a major source of reactive oxygen species (ROS), particularly during resting or state 4 respiration wherein ATP is not generated. One proposed role for respiratory mitochondrial uncoupling proteins (UCPs) is to decrease mitochondrial membrane potential and thereby protect cells from damage due to ROS. This work was designed to examine superoxide production during state 4 (no ATP production) and state 3 (active ATP synthesis) respiration and to determine whether uncoupling reduced the specific production of this radical species, whether this occurred in endothelial mitochondria per se, and whether this could be modulated by UCPs. Superoxide formation by isolated bovine aortic endothelial cell (BAE) mitochondria, determined using electron paramagnetic resonance spectroscopy, was approximately fourfold greater during state 4 compared with state 3 respiration. UCP1 and UCP2 overexpression both increased the proton conductance of endothelial cell mitochondria, as rigorously determined by the kinetic relationship of respiration to inner membrane potential. However, despite uncoupling, neither UCP1 nor UCP2 altered superoxide formation. Antimycin, known to increase mitochondrial superoxide, was studied as a positive control and markedly enhanced the superoxide spin adduct in our mitochondrial preparations, whereas the signal was markedly impaired by the powerful chemical uncoupler p-(trifluoromethoxyl)-phenyl-hydrazone. In summary, we show that UCPs do have uncoupling properties when expressed in BAE mitochondria but that uncoupling by UCP1 or UCP2 does not prevent acute substrate-driven endothelial cell superoxide as effluxed from mitochondria respiring in vitro. PMID- 15339749 TI - Professional liability crisis: a road map to success. PMID- 15339750 TI - Primary prevention of breast cancer: whither tamoxifen? PMID- 15339751 TI - Tamoxifen for breast cancer prevention: a framework for clinical decisions. AB - OBJECTIVE: Given the potential side effects and an uncertain survival benefit, decisions about tamoxifen treatment for the primary prevention of breast cancer remain complex. Primary care providers, including gynecologists, will need to counsel patients regarding this form of preventive care. In this report, we update cost-effectiveness calculations for tamoxifen chemoprevention and establish reasonable parameters for clinicians' use. METHODS: We performed a cost effectiveness analysis that compared women aged 50 years who were treated with tamoxifen for 5 years with an untreated cohort. In the base model, we assumed a 3.4% 5-year breast cancer risk. Quality-of-life estimates for important outcomes (breast cancer, endometrial cancer, deep venous thrombosis, pulmonary embolism, stroke, metastatic cancer, and hot flushes) were obtained from 106 women. Probabilities and costs of outcomes were derived from the Breast Cancer Chemoprevention Trial and other published estimates. Broad sensitivity analyses were performed. Cost per quality-adjusted life-year gained as a result of tamoxifen breast cancer prevention was the main outcome measure. RESULTS: The use of tamoxifen led to a remaining life expectancy of 26.07 quality-adjusted life years compared with 25.97 without treatment. The cost per quality-adjusted life year gained was $43,300. Sensitivity analysis revealed that younger age, the absence of the uterus, higher initial risk of breast cancer, increased fear of curable breast cancer, and reduced tamoxifen cost further favored treatment. CONCLUSION: Tamoxifen chemoprevention is cost-effective for women aged 40-50 years who are at significant breast cancer risk. Whether this holds true for older women depends on the initial breast cancer risk, fear of breast cancer, and presence of the uterus. PMID- 15339752 TI - Effects of ultralow-dose transdermal estradiol on bone mineral density: a randomized clinical trial. AB - OBJECTIVE: Because small increments in levels of endogenous plasma estradiol are associated with higher postmenopausal bone mineral density, we investigated the safety and effectiveness in preventing bone loss of unopposed, very-low-dose transdermal estradiol for postmenopausal women. METHODS: This was a randomized, placebo-controlled, double-blind trial with 2-year follow-up at 9 United States clinical centers. The study population comprised 417 postmenopausal women, aged 60-80 years, with intact uterus and bone mineral density z scores of -2.0 or higher, who were randomly assigned to receive either unopposed transdermal estradiol at 0.014 mg/d (n = 208) or placebo (n = 209). All participants received calcium and vitamin D supplementation. Lumbar spine and total hip bone mineral density change was measured by dual-energy X-ray absorptiometry; endometrial hyperplasia incidence was assessed by endometrial biopsy. RESULTS: Median plasma estradiol level in the estradiol group increased from 4.8 pg/mL at baseline to 8.5 pg/mL at 1 year (P <.001 versus baseline) and to 8.6 pg/mL at 2 years (P <.001 versus baseline) and was unchanged in the placebo group. Lumbar spine bone mineral density increased 2.6% in the estradiol group and 0.6% in the placebo group (between-group difference 2.0%, P <.001). Mean total hip bone mineral density increased 0.4% in the estradiol group and decreased 0.8% in the placebo group (between-group difference 1.2%, P <.001). Osteocalcin levels and bone specific alkaline phosphatase were lower in the estradiol group than the placebo group (P <.001 each). Endometrial hyperplasia developed in 1 woman in the estradiol group but in none of the placebo group (difference in 2-year rates 0.5%, 95% confidence interval 0-7.3%). CONCLUSION: Postmenopausal treatment with low-dose, unopposed estradiol increased bone mineral density and decreased markers of bone turnover without causing endometrial hyperplasia. PMID- 15339753 TI - Prevalence and characteristics of irritable bowel syndrome among women with chronic pelvic pain. AB - OBJECTIVE: We sought to evaluate whether there are unique characteristics associated with irritable bowel syndrome (IBS) within a population that has chronic pelvic pain. METHODS: This cross-sectional study of new referral patients attending a pelvic pain clinic between 1993 and 2000 (N = 987) evaluated characteristics associated with IBS at entry to the clinic. The characteristics that we evaluated included demographic characteristics, clinical diagnoses, history of abuse, depression, pain, and prior abdominal surgeries. RESULTS: Thirty-five percent of chronic pelvic pain patients had IBS defined by Rome I criteria. Age 40 years or older (odds ratio [OR] = 1.98, 95% confidence interval [CI]: 1.27, 3.11), muscular back pain (OR = 5.37, 95% CI: 0.98, 29.29), Symptom Checklist-90 global index score in top quartile (OR = 1.77, 95% CI: 1.09, 2.86), depression (OR = 1.93, 95% CI: 1.24, 3.01), 6 or more pain sites (OR = 1.67, 95% CI: 1.01, 2.78), and history of adult physical abuse (OR = 1.51, 95% CI: 1.01, 2.26) were associated with IBS in the final reduced multivariable model. CONCLUSION: Specific characteristics distinguish women with IBS suggesting that IBS and chronic pelvic pain are not simply manifestations of the same disorder. Our findings could help physicians attempt to effectively treat women with IBS and chronic pelvic pain. Physicians could diagnose and treat IBS in conjunction with treatment for chronic pelvic pain. PMID- 15339754 TI - Health, sociodemographic data, and pregnancy outcome in women with antepartum depressive symptoms. AB - OBJECTIVE: To study whether women with antepartum depression have an increased risk for adverse perinatal outcome. METHODS: From a sample of 1,489 women, an index group (n = 259) of all women with depressive symptoms on the Edinburgh Postnatal Depression Scale in gestational week 35-36 was selected. Two hundred fifty-nine women with no depressive symptoms on the Edinburgh Postnatal Depression Scale antepartum or postpartum were randomly chosen as the reference group. Medical, gynecologic, and obstetric history, socioeconomic status, pregnancy, and perinatal data were collected from standardized medical records for all women. RESULTS: Women with antepartum depressive symptoms were more often multiparas with a history of earlier obstetric complications. Complications during the present pregnancy were more frequent in the antepartum-depressed group of women. There were no differences concerning outcome of delivery, puerperium, and neonatal health between the index and reference groups. Forty-six percent of the women with antepartum depressive symptoms had depressive symptoms at 6-8 weeks or 6 months postpartum or both. CONCLUSION: Women depressed during pregnancy constitute a group without an increased risk for adverse obstetric or neonatal outcome but with a high risk for postpartum depressive symptoms. PMID- 15339755 TI - Implications of antenatal depression and anxiety for obstetric outcome. AB - OBJECTIVE: To investigate the obstetric outcome and health care consumption during pregnancy, delivery, and the early postpartum period in an unselected population-based sample of pregnant women diagnosed with antenatal depressive and/or anxiety disorders, compared with healthy subjects. METHODS: Participants were 1,495 women attending 2 obstetric clinics in Northern Sweden. The Primary Care Evaluation of Mental Disorders was used to evaluate depressive and anxiety disorders in the second trimester of pregnancy. To assess demographic characteristics, obstetric outcome, and complications, the medical records of the included women were reviewed. RESULTS: Significant associations were found between depression and/or anxiety and increased nausea and vomiting, prolonged sick leave during pregnancy and increased number of visits to the obstetrician, specifically, visits related to fear of childbirth and those related to contractions. Planned cesarean delivery and epidural analgesia during labor were also significantly more common in women with antenatal depression and/or anxiety. CONCLUSION: There is an association between antenatal depressive and/or anxiety disorders and increased health care use (including cesarean deliveries) during pregnancy and delivery. PMID- 15339756 TI - Mental and physical distress and high-risk behaviors among reproductive-age women. AB - OBJECTIVE: To examine the prevalence of mental and physical distress indicators among women of reproductive age and the association of these indicators with cigarette smoking and alcohol use, by pregnancy status. METHODS: The Behavioral Risk Factor Surveillance System data for several years were aggregated across states and weighted for this analysis. Seven measures of self-reported mental and physical distress and general health were examined along with demographic variables. RESULTS: Overall, 6.7% (95% confidence interval [CI] 6.5-6.9) of women reported frequent physical distress, 12.3% (95% CI 12.0-12.6) reported frequent mental distress, 9.9% (95% CI 9.4-10.4) reported frequent depression, 18.4% (95% CI 17.8-19.1) reported feeling anxious, and 34.3% (95% CI 33.5-35.1) reported that they frequently did not get enough rest. At the time of the survey 4.6% of the women were pregnant. Pregnant women were less likely than nonpregnant women to report frequent mental distress. Although there was attenuation of cigarette smoking and alcohol use during pregnancy, those with mental and physical distress were more likely to consume cigarettes and alcohol than were those without such experiences. CONCLUSION: High proportions of reproductive-age women report frequent mental and physical distress. Women experiencing mental and physical distress were more likely to report consuming cigarettes and alcohol than women without such experiences. PMID- 15339757 TI - Short-term outcomes in low birth weight infants following antenatal exposure to betamethasone versus dexamethasone. AB - OBJECTIVE: To assess the incidence of short-term outcomes of low birth weight infants ( 24 hours), and the number of women who had received tocolysis. No significant difference was found between the 2 groups with respect to intraventricular hemorrhage and cystic PVL frequencies. No significant differences were found in the incidence of short-term outcomes examined, despite the fact that the dexamethasone group was exposed to a statistically significantly greater number of courses than the betamethasone group. CONCLUSION: There seem to be no advantages to maternal antenatal treatment with betamethasone compared with dexamethasone in reducing the risk of PVL in low birth weight (/= 10 weeks of gestation) and having at least one subsequent pregnancy. Patients with antiphospholipid antibodies were excluded. Logistic regression analysis was performed to determine the association of clinical variables with pregnancy outcome. RESULTS: Two hundred thirty subjects met inclusion criteria. Up through the time of their first fetal death, these women had a total of 721 pregnancies, resulting in 268 (37%) live births, 230 (32%) fetal deaths, and 200 (28%) spontaneous abortions. In total, these women had 839 subsequent pregnancies, resulting in 202 (24%) live births, 209 (25%) fetal deaths, and 372 (44%) spontaneous abortions. Univariate logistic regression analysis identified older age at pregnancy (P =.009, odds ratio 0.63, 95% confidence interval 0.42-1.03) and treatment with low-dose aspirin (P =.001, odds ratio 0.41, 95% confidence interval 0.25-0.68) to be associated with a decreased risk for subsequent pregnancy loss. CONCLUSION: Women with prior fetal death are at high risk for subsequent pregnancy loss and recurrent fetal death, with fewer than 25% of pregnancies resulting in surviving infants. These data underscore the need for additional research into the pathophysiology and prevention of recurrent fetal death. PMID- 15339763 TI - Perinatal outcome of pregnancies complicated by placenta accreta. AB - OBJECTIVE: The purpose of the study was to characterize the perinatal outcome of pregnancies complicated by placenta accreta. METHODS: We conducted a case-control analysis of all deliveries between the years 1990 and 2000 that were complicated by placenta accreta. Perinatal variables included in the analysis were gestational age at delivery, birth weight, Apgar scores, and perinatal mortality. Statistical analysis was performed using both the unpaired and paired approach. P <.05 was considered significant. RESULTS: The study encompassed 34,450 deliveries, from which 310 cases of placenta accreta were diagnosed (0.9%) and compared with 310 matched controls. In the pregnancies complicated by placenta accreta, we found a statistically significant increase in preterm deliveries (10.7% versus 1%, P <.001, odds ratio 12.1, 95% confidence interval 3.7-39.9) and small-for-gestational-age babies (27.3% versus 14%, P <.001, odds ratio 5.05, 95% confidence interval 1.46-3.28). CONCLUSION: Pregnancies complicated by placenta accreta are at increased risk for perinatal adverse outcome. We speculate that these findings may arise from pathological implantation of the placenta, resulting in interference with normal fetal growth. PMID- 15339764 TI - Conservative versus extirpative management in cases of placenta accreta. AB - OBJECTIVE: To compare the impact of conservative and extirpative strategies for placenta accreta on maternal morbidity and mortality. METHODS: We retrospectively reviewed the medical records of all patients diagnosed with placenta accreta admitted to our tertiary center from January 1993 through December 2002. Two consecutive periods, A and B, were compared. During period A (January 1993 to June 1997), our written protocol called for the systematic manual removal of the placenta, to leave the uterine cavity empty. In period B (July 1997 to December 2002), we changed our policy by leaving the placenta in situ. The following outcomes over the 2 periods were compared: need for blood transfusion, hysterectomy, intensive care admission, duration of stay in intensive care, and postpartum endometritis. RESULTS: Thirty-three cases of placenta accreta were observed among 31,921 deliveries (1.03/1,000). During period B, there was a reduction in the hysterectomy rate (from 11 [84.6%] to 3 [15%]; P <.001), the mean number of red blood cells transfused (3,230 +/- 2,170 mL versus 1,560 +/- 1,646 mL; P <.01), and disseminated intravascular coagulation (5 [38.5%] versus 1 [5.0%]; P =.02), compared with period A. There were 3 cases of sepsis in period B and none in period A (P =.26). At least 2 women with conservative management subsequently had successful pregnancies. CONCLUSION: Leaving the placenta accreta in situ appears to be a safe alternative to removing the placenta. PMID- 15339765 TI - Prenatal diagnosis of placenta accreta and percreta with ultrasonography, color Doppler, and magnetic resonance imaging. AB - BACKGROUND: The risk of placenta previa and accreta is increased in females with previous cesarean deliveries, and there has been an increasing number of these operations. CASES: We present 2 cases with previous cesarean and placenta previa in the following pregnancy. One patient had placenta accreta and the other, placenta percreta. In both cases, prenatal diagnosis was based on ultrasonography, where features such as loss of the hypoechoic retroplacental zone and irregular uterine serosa were found in grayscale ultrasonography. In color Doppler imaging, in both cases, increased vascularity between myometrium and placenta, as well as intraplacental lacunae, were seen. Thinning of the uterine wall, found in magnetic resonance imaging, contributed to the diagnosis of placenta percreta. CONCLUSION: Prenatal diagnosis of placenta accreta is of importance because it reduces fetal and maternal morbidity as appropriate preoperative and perioperative procedures are possible. PMID- 15339766 TI - Responses by pregnant Jehovah's Witnesses on health care proxies. AB - OBJECTIVE: To review the treatment options presented on the New York State Health Care Proxy for Jehovah's Witnesses, which is signed by pregnant women when they present for care. METHODS: Chart reviews were performed for all women who presented to labor and delivery at our institution from 1997 to 2002 and identified themselves as Jehovah's Witnesses. A patient was included in the study if a completed health care proxy was available in her chart. Data were derived from the health care proxy and from the Mount Sinai School of Medicine's Blood Product Checklist for Jehovah's Witness Patients. Variables of interest included age, race, parity, and antenatal and perinatal complications. RESULTS: A total of 61 patients were identified. Of these, 39.3% agreed to accept a variety of donated blood products, 9.8% would accept donated packed red blood cells, and 50.1% would accept neither from a homologous donor. With respect to nonstored autologous blood, 55% of respondents would accept either intraoperative normovolemic hemodilution or transfusion of their own blood obtained by a cell salvage system. No significant differences in responses were noted for any of the above-mentioned variables. CONCLUSION: This review refutes the commonly held belief that all Jehovah's Witnesses refuse to accept blood or any of its products. In this population of pregnant women, the majority were willing to accept some form of blood or blood products. This information can be used to help health care providers counsel a patient when she is initially faced with considering these issues and may help to remove the stigma of accepting one of the options. PMID- 15339767 TI - Inverse correlation between maternal weight and second trimester circulating cell free fetal DNA levels. AB - OBJECTIVE: Clinical applications of the analysis of cell-free fetal DNA in maternal plasma and serum are expanding. However, use of fetal DNA during prenatal screening requires knowledge of variables that might affect its levels in the maternal circulation. We conducted this study to estimate the effect of selected demographic factors on fetal DNA levels in the first and second trimesters of pregnancy. METHODS: We developed a database that included fetal DNA levels and clinical information, such as maternal age, ethnicity, weight, and smoking history. We measured fetal DNA levels in maternal plasma and serum using real-time quantitative polymerase chain reaction amplification of a Y chromosome specific sequence. The fetal DNA data from fresh first trimester plasma and previously frozen second trimester serum samples were analyzed separately. Fetal DNA levels were adjusted according to gestational age and storage time and then analyzed in association with the demographic factors. RESULTS: In the first trimester group, no significant association between maternal age, weight, ethnic background, or smoking and plasma fetal DNA levels was observed. In the second trimester group, a significant inverse correlation between maternal weight and serum fetal DNA level was demonstrated (r = -0.26, P =.007). This was especially prominent when the mothers weighed more than 170 lb (P =.001). Maternal age, ethnicity, and smoking were not significantly associated with the second trimester serum fetal DNA levels. CONCLUSION: Fetal DNA levels are affected by maternal weight in the second trimester. A correction for this effect may be needed in larger-scale studies or for future clinical applications that measure cell-free fetal nucleic acids in maternal circulation. PMID- 15339768 TI - Low risk of relapse after achieving undetectable HCG levels in women with complete molar pregnancy. AB - OBJECTIVE: Complete hydatidiform molar pregnancies occur in approximately 1 of 1,000 conceptions. After uterine evacuation of the trophoblastic tissue, women are followed up with serial serum human chorionic gonadotropin (hCG) measurements. Patients are considered to have attained remission when their hCG level spontaneously declines to an undetectable level and remains there during a 6-month follow-up period. This standard effectively detects all disease recurrence; however, it is resource intensive, delays child bearing, and is subject to significant noncompliance. Our objective was to determine the risk of disease recurrence after hCG spontaneously declines to undetectable levels. METHODS: We used a database from the New England Trophoblastic Disease Center to analyze hCG levels in patients with complete molar pregnancies. RESULTS: Among 1,029 women with complete molar pregnancy and complete data, 15% developed persistent gestational trophoblastic neoplasia. The rate of persistent neoplasm among those whose hCG level fell spontaneously to undetectable levels was 0.2% (2/876, 95% confidence interval 0-0.8%). No women developed persistent gestational trophoblastic neoplasia after their hCG level fell to undetectable levels using an assay with a sensitivity of 5 mIU/mL (n = 82, 95% confidence interval 0-4.5%). CONCLUSION: Based on our experience with women with complete hydatidiform molar pregnancies whose hCG values spontaneously fell to undetectable levels after molar evacuation, we conclude that the risk of recurrent neoplasm after hCG levels fall to less than 5 mIU/mL approaches zero. PMID- 15339769 TI - A comparison between the vaginal ring and oral contraceptives. AB - OBJECTIVE: To investigate whether contraceptive vaginal ring use results in similar estimated genital symptoms, signs, examination, and laboratory findings compared with oral contraceptive use. METHODS: Women were randomly assigned to either contraceptive vaginal ring or a 20 microg ethinyl estradiol oral contraceptive pill use for 3 consecutive 28-day cycles, directly followed by 3 cycles of the study drug not initially assigned. Subjects scored genital symptoms on a daily diary using a 0-4 scale and underwent a baseline, cycle 2, cycle 4, and exit pelvic examination including vaginal discharge evaluation, vaginal Gram stain and white cell count, and culture for yeast and Lactobacillus, including colony count and hydrogen peroxide production. RESULTS: Of the 40 subjects assigned to each arm, 33 (82.5%) subjects in the ring-first arm and 31 (77.5%) subjects in the pill-first arm completed all study visits (P =.58). Most subjects reported few genital symptoms with either method, but 63% of subjects reported vaginal wetness during ring use compared with 43% during pill use. During ring use larger numbers of Lactobacillus colonies present were positive for hydrogen peroxide production (fold difference 2.67, 95% confidence interval 1.49, 4.78, P <.001). All other laboratory data, including yeast colony counts, Nugent Gram stain score, vaginal white blood cell count, vaginal pH, and discharge weight, were not significantly different by method. CONCLUSION: Some women may notice an increase in vaginal wetness during contraceptive ring use yet the method is well tolerated and appears to improve the vaginal flora. PMID- 15339770 TI - Interleukin-6 and antiphospholipid antibodies in women with contraceptive-related thromboembolic disease. AB - OBJECTIVE: The aim of this study was to explore the possible (joint) contributing role of interleukin-6 (IL-6) and antiphospholipid antibodies to the occurrence of the venous thromboembolism in women using oral contraceptives. METHODS: Interleukin-6 and antiphospholipid antibodies (anti-beta2-glycoprotein I antibody immunoglobulin M [IgM], G [IgG], and A [IgA]; anticardiolipin-IgM and IgG; antiphosphatidylserine-IgM and IgG) were measured in 30 women (median age 41, range 28-49 years) in the stable period (on average 3.5 years) after first venous thromboembolism. Sixteen patients used oral contraceptives during the episode of venous thromboembolism (oral contraceptives group), whereas 14 patients did not (non-oral contraceptives group). Thirty-seven age-matched, healthy women served as controls RESULTS: Compared with controls, the oral contraceptives group had elevated IL-6 (median interquartile range 2.3 [1.1-4.3] versus 1.4 [0-2.0] pg/mL, P <.05). The oral contraceptives group had elevated anti-beta2-glycoprotein I antibody-IgM in comparison with both the non-oral contraceptives group (median interquartile range 47.5 [2.0-77.0] versus 29.50 [11.00-45.50] OD(450), P <.06) and controls (47.5 [2.0-77.0] versus 17.5 [3.5-30.0] OD(450), P <.001). Interleukin-6 level in the non-oral contraceptives group was related to obesity, whereas such a relation was not found in the oral contraceptives group, suggesting the presence of another factor (oral contraceptive use), which stimulates IL-6 production. Of particular interest is our finding that elevated IL-6 levels correlated significantly positively with elevated anti-beta2 glycoprotein I antibody-IgG in patients who were users of oral contraceptives (but not overweight, n = 10) (r = 0.56, P <.05) CONCLUSION: The results suggest a new hypothesis that, in susceptible women, use of oral contraceptives induces production of IL-6, which stimulates production of anti-beta2-glycoprotein I. Thus, the prothrombotic profile is aggravated and could facilitate occurrence of venous thromboembolism. This remains to be elucidated in further studies. PMID- 15339771 TI - Accuracy of transvaginal ultrasonography in diabetic or obese women with postmenopausal bleeding. AB - OBJECTIVE: We sought to assess the accuracy of endometrial thickness measurement in the diagnosis of endometrial cancer in patients with obesity, diabetes, and hypertension and to evaluate whether patient characteristics influence endometrial thickness irrespective of the final diagnosis. METHODS: This was a prospective study of women not using hormone replacement therapy who presented with postmenopausal bleeding at 8 hospitals in The Netherlands. All women underwent transvaginal ultrasonography and, in the event that the endometrial thickness (double layer) was more than 4 mm, subsequent endometrial sampling. The performance of endometrial thickness measurement in the diagnosis of atypical hyperplasia and endometrial cancer was evaluated in subgroups of patients with diabetes, hypertension, and obesity by using receiver operating characteristic analysis. RESULTS: Overall, we included 594 consecutive women, of whom 62 (10%) had endometrial carcinoma and 6 (1%) had atypical hyperplasia. In these women, transvaginal ultrasonography had an area under the receiver operating characteristic curve of 0.87 (standard error [SE] 0.03). In the absence of (pre)malignancy, women with diabetes or obesity were found to have thicker endometria than women without these risk factors, whereas in women with a (pre)malignancy, this difference was not present. The area under the receiver operating characteristic curve decreased to 0.74 (SE 0.05) and 0.75 (SE 0.07) in diabetic women and obese women, respectively. The presence or absence of hypertension had no impact on the accuracy of transvaginal ultrasonography. CONCLUSION: In view of the decreased diagnostic accuracy in diabetic women and obese women, the clinical value of transvaginal endometrial thickness measurement in these women is questionable. PMID- 15339772 TI - Misoprostol outpatient cervical ripening without subsequent induction of labor: a randomized trial. AB - OBJECTIVE: To estimate the effect of outpatient administration of a single dose of vaginal misoprostol at term on the interval to delivery in women with unfavorable cervices. METHODS: Randomized, double blind, placebo-controlled trial comparing a single 25-microg outpatient intravaginal dose of misoprostol to placebo in pregnant women with Bishop scores less than 9 at 40 weeks or greater. After placement of the study medication, subjects were permitted to go into spontaneous labor unless an indication for induction developed. Analysis was by intent to treat. The interval to delivery, defined as the time from medication placement to delivery, was compared by Student t test and by survival analysis with the log-rank test. RESULTS: Thirty-three women were randomly assigned to receive misoprostol, and 35 were assigned to receive placebo. The mean interval to delivery was significantly less in the misoprostol group, 4.2 +/- 4.1 compared with 6.1 +/- 3.6 days, P =.04. The interval to delivery for only the nulliparous patients was significantly less in the misoprostol group, 4.2 +/- 4.0 compared with 7.2 +/- 3.7 days, P =.02. The survival curves for the interval to delivery were significantly different (P =.04 by log-rank test) with 4.1 days median interval to delivery for misoprostol compared with 9.2 days for placebo. There were no adverse outcomes in either group. CONCLUSION: A single 25-microg outpatient intravaginal dose of misoprostol is effective in decreasing the interval to delivery in women with unfavorable cervices at term. PMID- 15339773 TI - Association of asthma diagnosis, severity, symptoms, and treatment with risk of preeclampsia. AB - OBJECTIVE: Existing studies relating asthma and preeclampsia provide conflicting results, perhaps due to differences in study populations, varying definitions of asthma, and inadequate control for confounding, particularly asthma medication use. This prospective study examines associations between aspects of asthma (diagnosis, severity, symptoms, and medication use) and risk of preeclampsia. METHODS: A total of 1,708 pregnant women, of whom 656 had asthma diagnosis and 1,052 had no asthma diagnosis, were included in this analysis. Asthma symptoms, treatment, and severity were classified according to Global Initiative for Asthma guidelines. Hospital records were abstracted, and strict criteria were applied to classify women as preeclamptic based on National Heart, Lung, and Blood Institute guidelines. RESULTS: There were 568 of 656 women with diagnosed asthma and 353 of 1,052 women without asthma diagnosis who had symptoms or took asthma medication during pregnancy. Separate adjusted logistic regression models were run for different measures of asthma status: 1) asthma diagnosis; 2) overall Global Initiative for Asthma severity; 3) Global Initiative for Asthma symptom and treatment steps; and 4) Global Initiative for Asthma symptom step and medication type. Women at increased risk of preeclampsia were those classified as Global Initiative for Asthma symptom step 3/4 compared with no symptoms (odds ratio 3.36, 95% confidence interval 1.24-9.14) and theophylline users (odds ratio 1.16 for every dose/month increase in use, 95% confidence interval 1.02-1.33). In contrast, neither a history of physician-diagnosed asthma nor Global Initiative for Asthma treatment step was associated with preeclampsia status. CONCLUSION: Our findings suggest that women with moderate to severe asthma symptoms, regardless of asthma diagnosis or treatment, are at increased risk of preeclampsia compared with women with no symptoms. PMID- 15339774 TI - Transmission of parentally shared human leukocyte antigen alleles and the risk of preterm delivery. AB - OBJECTIVE: Our objective was to examine our hypothesis that the transmission of parentally shared human leukocyte antigen (HLA) alleles to offspring increases the risk of preterm delivery. METHODS: A population-based family study with participating children and their parents was conducted in Kaiser Permanente Medical Care Program, an integrated healthcare delivery system, in the Northern California Region. A total of 234 participants from 78 families with early preterm deliveries (35 weeks of gestation or greater) and 60 participants from 20 families with full-term births were included in the study. Buccal cells were collected from the first-born preterm cases and their parents to determine HLA-B (class I) and DRB1 (class II) types and the transmission of parental alleles to the offspring. The buccal samples were also collected from full-term deliveries to rule out possible segregation distortion at the studied HLA loci. RESULTS: Compared with the expected transmission probability based on Mendel's laws (25%), transmission of parentally shared HLA-B or DRB1 alleles from both heterozygous parents to offspring (48% of 23 heterozygous parents) was associated with a more than 5-fold increased risk of preterm delivery (odds ratio 5.5; 95% confidence interval 1.2-51). Transmission of parentally shared HLA alleles from heterozygous mothers (83%) appears to be more important in the etiology of preterm delivery than transmission from fathers (57%). The transmission pattern of parentally shared HLA alleles in our full-term controls was almost identical to the expected pattern based on Mendel's laws and demonstrated no segregation distortion at those HLA loci. CONCLUSION: Our findings provide evidence that the transmission of parentally shared HLA alleles may be an underlying mechanism for preterm delivery. PMID- 15339775 TI - Estimating blood loss: can teaching significantly improve visual estimation? AB - OBJECTIVE: Visually estimated blood loss has long been known to be imprecise, inaccurate, and often underestimated, which may lead to delayed diagnosis and treatment. Our purpose is to determine whether a brief didactic course can improve visually estimated blood loss and whether prior clinical experience influences estimation of blood loss. METHODS: Reconstituted whole blood was obtained from the blood bank, and simulated scenarios with known measured blood loss were created using common surgical materials. Visually estimated blood loss was performed by medical personnel before and after a 20-minute didactic session. Percent errors of estimated blood loss were calculated and comparisons were made before and after the lecture. The effects of actual blood volume and clinical experience on estimation of blood loss were assessed. RESULTS: A total of 53 participants assessed 7 scenarios. There were significant reductions in error for all scenarios. Median percent error in estimated blood loss was not influenced by clinical experience, either before or after the didactic session. Blood loss tends to be overestimated at low volumes and underestimated at high volumes. CONCLUSION: Error in estimating blood loss is dependent on actual blood loss volume. Medical students and experienced faculty demonstrate similar errors, and both can be improved significantly with limited instruction. This educational process may assist clinicians in everyday practice to more accurately estimate blood loss and recognize patients at risk for hemorrhage-related complications. PMID- 15339776 TI - Stress urinary incontinence. AB - Stress urinary incontinence, the complaint of involuntary leakage during effort or exertion, occurs at least weekly in one third of adult women. The basic evaluation of women with stress urinary incontinence includes a history, physical examination, cough stress test, voiding diary, postvoid residual urine volume, and urinalysis. Formal urodynamics testing may help guide clinical care, but whether urodynamics improves or predicts the outcome of incontinence treatment is not yet clear. The distinction between urodynamic stress incontinence associated with hypermobility and urodynamic stress incontinence associated with intrinsic sphincter deficiency should be viewed as a continuum, rather than a dichotomy, of urethral function. Initial treatment should include behavioral changes and pelvic floor muscle training. Estrogen is not indicated to treat stress urinary incontinence. Bladder training, vaginal devices, and urethral inserts also may reduce stress incontinence. Bulking agents reduce leakage, but effectiveness generally decreases after 1-2 years. Surgical procedures are more likely to cure stress urinary incontinence than nonsurgical procedures but are associated with more adverse events. Based on available evidence at this time, colposuspension (such as Burch) and pubovaginal sling (including the newer midurethral synthetic slings) are the most effective surgical treatments. PMID- 15339777 TI - Impose legal limits to preserve freedoms. PMID- 15339778 TI - Is it time to take a "REST"? PMID- 15339780 TI - Elective cesarean delivery. PMID- 15339781 TI - In-flight radiation exposure during pregnancy. PMID- 15339783 TI - Maternal morbidity associated with vaginal versus cesarean delivery. PMID- 15339785 TI - Improving resident competency in the management of shoulder dystocia with simulation training. PMID- 15339787 TI - Risk factors for legal induced abortion-related mortality in the United States. PMID- 15339790 TI - ACOG Practice Bulletin. Clinical management guidelines for obstetricians gynecologists. Number 55, September 2004 (replaces practice pattern number 6, October 1997). Management of Postterm Pregnancy. PMID- 15339791 TI - ACOG Committee Opinion. Number 299, September 2004 (replaces No. 158, September 1995). Guidelines for diagnostic imaging during pregnancy. PMID- 15339792 TI - Vascular endothelial growth factor and nephrin interact and reduce apoptosis in human podocytes. AB - Vascular endothelial growth factor (VEGF) is anti-cytotoxic in podocytes. Moreover, it has been suggested that nephrin, a cell adhesion molecule of the podocyte slit diaphragm, can contribute to antiapoptotic mechanisms in these cells. We therefore investigated whether VEGF signals to reduce apoptosis and the role of nephrin in this survival mechanism. Flow cytometry showed that podocytes with nephrin mutations had a significantly greater proportion of apoptosis. Although VEGF reduced apoptosis in human conditionally immortalized podocytes [wild-type (WT)] by 18.1% of control (P < 0.001), it was unable to do so in nephrin-deficient human conditionally immortalized podocytes. Moreover, Western blotting and immunodetection with an anti-nephrin antibody showed that the phosphorylation of nephrin, compared with serum-starved WTs, was significantly increased (ratio of 3.36 +/- 1.2 to control, P < 0.05) by VEGF treatment and significantly reduced by treatment with a neutralizing VEGF monoclonal antibody (mAb) (ratio of 0.2 +/- 0.09 to control, P < 0.05). The AKT signaling pathway has been implicated in nephrin-mediated inhibition of apoptosis in transfected cells, but the role of this pathway has not previously been shown in podocytes. Surprisingly, exogenous VEGF decreased AKT/PKB phosphorylation in normal podocytes but increased it in nephrin-deficient podocytes. We suggest therefore that both exogenous and endogenous (podocyte derived) VEGF can stimulate the phosphorylation of nephrin and through this action may prevent podocyte apoptosis. However, the involvement of AKT in this survival response in normal human podocytes is not clear. PMID- 15339793 TI - Cd(2+)-induced cytochrome c release in apoptotic proximal tubule cells: role of mitochondrial permeability transition pore and Ca(2+) uniporter. AB - Cd(2+) induces apoptosis of kidney proximal tubule (PT) cells. Mitochondria play a pivotal role in toxic compound-induced apoptosis by releasing cytochrome c. Our objective was to investigate the mechanisms underlying Cd(2+)-induced cytochrome c release from mitochondria in rat PT cells. Using Hoechst 33342 or MTT assay, 10 muM Cd(2+) induced approximately 5-10% apoptosis in PT cells at 6 and 24 h, which was associated with cytochrome c and apoptosis-inducing factor release at 24 h only. This correlated with previously described maximal intracellular Cd(2+) concentrations at 24 h, suggesting that elevated Cd(2+) may directly induce mitochondrial liberation of proapoptotic factors. Indeed, Cd(2+) caused swelling of energized isolated kidney cortex mitochondria (EC(50) approximately 9 muM) and cytochrome c release, which were independent of permeability transition pore (PTP) opening since PTP inhibitors cyclosporin A or bongkrekic acid had no effect. On the contrary, Cd(2+) inhibited swelling and cytochrome c release induced by PTP openers (PO(4)(3-) or H(2)O(2)+Ca(2+)). The mitochondrial Ca(2+) uniporter (MCU) played a key role in mitochondrial damage: 1) MCU inhibitors (La(3+), ruthenium red, Ru360) prevented swelling and cytochrome c release; and 2) ruthenium red attenuated Cd(2+) inhibition of PO(4)(3-)-induced swelling. Using the Cd(2+)-sensitive fluorescent indicator FluoZin-1, Cd(2+) was also taken up by mitoplasts. The aquaporin inhibitor AgNO(3) abolished Cd(2+)-induced swelling of mitoplasts. This could be partially mediated by activation of the mitoplast-enriched water channel aquaporin-8. Thus cytosolic Cd(2+) concentrations exceeding a certain threshold may directly cause mitochondrial damage and apoptotic development by interacting with MCU and water channels in the inner mitochondrial membrane. PMID- 15339794 TI - Both Sp1 and Smad participate in mediating TGF-beta1-induced HGF receptor expression in renal epithelial cells. AB - Hepatocyte growth factor (HGF) receptor is a transmembrane receptor tyrosine kinase encoded by the c-met protooncogene. In this study, we demonstrated that c met expression was upregulated in the kidney after obstructive injury in mice. Because the pattern of c-met induction was closely correlated with transforming growth factor-beta1 (TGF-beta1) expression in vivo, we further investigated the regulation of c-met expression in renal tubular epithelial (HKC) cells by TGF beta1 in vitro. Real-time RT-PCR and Northern and Western blot analyses revealed that TGF-beta1 significantly induced c-met expression in HKC cells, which primarily took place at the gene transcriptional level. Overexpression of inhibitory Smad7 completely abolished c-met induction, indicating its dependence on Smad signaling. Interestingly, TGF-beta1-induced c-met expression was also contingent on a functional Sp1, as ablation of Sp1 binding with mithramycin A abrogated c-met induction in HKC cells. Transfection and sequence analysis identified a cis-acting TGF-beta1-responsive region in the c-met promoter, in which resided a putative Smad-binding element (SBE) and an adjacent Sp1 site. TGF beta1 not only induced Smad binding to the SBE/Sp1 sites in the c-met promoter, but also enhanced the binding of Sp proteins. Furthermore, Sp1 could form a complex with Smads in a TGF-beta1-dependent fashion. These results suggest a novel regulatory mechanism controlling c-met expression by TGF-beta1 in renal epithelial cells, in which both Smad and Sp proteins participate and cooperate in activating c-met gene transcription. PMID- 15339795 TI - The protein-tethered lipid bilayer: a novel mimic of the biological membrane. AB - A new concept of solid-supported tethered bilayer lipid membrane (tBLM) for the functional incorporation of membrane proteins is introduced. The incorporated protein itself acts as the tethering molecule resulting in a versatile system in which the protein determines the characteristics of the submembraneous space. This architecture is achieved through a metal chelating surface, to which histidine-tagged (His-tagged) membrane proteins are able to bind in a reversible manner. The tethered bilayer lipid membrane is generated by substitution of protein-bound detergent molecules with lipids using in-situ dialysis or adsorption. The system is characterized by surface plasmon resonance, quartz crystal microbalance, and electrochemical impedance spectroscopy. His-tagged cytochrome c oxidase (CcO) is used as a model protein in this study. However, the new system should be applicable to all recombinant membrane proteins bearing a terminal His-tag. In particular, combination of surface immobilization and membrane reconstitution opens new prospects for the investigation of functional membrane proteins by various surface-sensitive techniques under a defined electric field. PMID- 15339796 TI - Dynamic structure of vesicle-bound melittin in a variety of lipid chain lengths by solid-state NMR. AB - Solid-state 31P- and 13C-NMR spectra were recorded in melittin-lecithin vesicles composed of 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC) or 1,2-dipalmitoyl sn-glycero-3-phosphocholine (DPPC). Highly ordered magnetic alignments were achieved with the membrane surface parallel to the magnetic field above the gel to-liquid crystalline phase transition temperature (Tc). Using these magnetically oriented vesicle systems, dynamic structures of melittin bound to the vesicles were investigated by analyzing the 13C anisotropic and isotropic chemical shifts of selectively 13C-labeled carbonyl carbons of melittin under the static and magic-angle spinning conditions. These results indicate that melittin molecules adopt an alpha-helical structure and laterally diffuse to rotate rapidly around the membrane normal with tilt angles of the N-terminal helix being -33 degrees and -36 degrees and those of the C-terminal helix being 21 degrees and 25 degrees for DLPC and DPPC vesicles, respectively. The rotational-echo double-resonance method was used to measure the interatomic distance between [1-13C]Val8 and [15N]Leu13 to further identify the bending alpha-helical structure of melittin to possess the interhelical angles of 126 degrees and 119 degrees in DLPC and DPPC membranes, respectively. These analyses further lead to the conclusion that the alpha-helices of melittin molecules penetrate the hydrophobic cores of the bilayers incompletely as a pseudo-trans-membrane structure and induce fusion and disruption of vesicles. PMID- 15339797 TI - Simulation of the early stages of nano-domain formation in mixed bilayers of sphingomyelin, cholesterol, and dioleylphosphatidylcholine. AB - It is known from experimental studies that lipid bilayers composed of unsaturated phospholipids, sphingomyelin, and cholesterol contain microdomains rich in sphingomyelin and cholesterol. These domains are similar to "rafts" isolated from cell membranes, although the latter are much smaller in lateral size. Such domain formation can be a result of very specific and subtle lipid-lipid interactions. To identify and study these interactions, we have performed two molecular dynamics simulations, of 200-ns duration, of dioleylphosphatidylcholine (DOPC), sphingomyelin (SM), and cholesterol (Chol) systems, a 1:1:1 mixture of DOPC/SM/Chol, and a 1:1 mixture of DOPC/SM. The simulations show initial stages of the onset of spontaneous phase-separated domains in the systems. On the simulation timescale cholesterol favors a position at the interface between the ordered SM region and the disordered DOPC region in the ternary system and accelerates the process of domain formation. We find that the smooth alpha-face of Chol preferentially packs next to SM molecules. Based on a comparative analysis of interaction energies, we find that Chol molecules do not show a preference for SM or DOPC. We conclude that Chol molecules assist in the process of domain formation and the process is driven by entropic factors rather than differences in interaction energies. PMID- 15339798 TI - Molecular dynamics study of gating in the mechanosensitive channel of small conductance MscS. AB - Mechanosensitive channels are a class of ubiquitous membrane proteins gated by mechanical strain in the cellular membrane. MscS, the mechanosensitive channel of small conductance, is found in the inner membrane of Escherichia coli and its crystallographic structure in an open form has been recently solved. By means of molecular dynamics simulations we studied the stability of the channel conformation suggested by crystallography in a fully solvated lipid (POPC) bilayer, the combined system encompassing 224,340 atoms. When restraining the backbone of the protein, the channel remained in the open form and the simulation revealed intermittent permeation of water molecules through the channel. Abolishing the restraints under constant pressure conditions led to spontaneous closure of the transmembrane channel, whereas abolishing the restraints when surface tension (20 dyn/cm) was applied led to channel widening. The large balloon-shaped cytoplasmic domain of MscS exhibited spontaneous diffusion of ions through its side openings. Interaction between the transmembrane domain and the cytoplasmic domain of MscS was observed and involved formation of salt bridges between residues Asp62 and Arg128; this interaction may be essential for the gating of MscS. K+ and Cl- ions showed distinctively different distributions in and around the channel. PMID- 15339799 TI - Membrane tether extraction from human umbilical vein endothelial cells and its implication in leukocyte rolling. AB - During the rolling of human neutrophils on the endothelium, tethers (cylindrical membrane tubes) are likely extracted from the neutrophil. Tether extraction reduces the force imposed on the adhesive bond between the neutrophil and endothelium, thereby facilitating the rolling. However, whether tethers can be extracted from the endothelium is still unknown. Here, with the micropipette aspiration technique, we show that tethers can be extracted from either suspended or attached human umbilical vein endothelial cells. We also show that a linear relationship between the pulling force and tether growth velocity exists and this relationship does not depend on the receptor type (used to impose point forces), tumor necrosis factor-alpha stimulation, or cell attachment state. With linear regression, we determined that the threshold force was 50 pN and the effective viscosity was 0.50 pN.s/microm. Therefore, tethers might be simultaneously extracted from the neutrophil and endothelial cell during the rolling and, more importantly, the endothelial cell might contribute much more to the total composite tether length than the neutrophil. Compared with tether extraction from the neutrophil alone, simultaneous tether extraction results in a larger increase in the lifetime of the adhesive bond, and thus further stabilizes the rolling of neutrophils under high physiological shear stresses. PMID- 15339800 TI - Long-residency hydration, cation binding, and dynamics of loop E/helix IV rRNA L25 protein complex. AB - Molecular dynamics simulations of RNA-protein complex between Escherichia coli loop E/helix IV (LE/HeIV) rRNA and L25 protein reveal a qualitative agreement between the experimental and simulated structures. The major groove of LE is a prominent rRNA cation-binding site. Divalent cations rigidify the LE major groove geometry whereas in the absence of divalent cations LE extensively interacts with monovalent cations via inner-shell binding. The HeIV region shows bistability of its major groove explaining the observed differences between x-ray and NMR structures. In agreement with the experiments, the simulations suggest that helix alpha1 of L25 is the least stable part of the protein. Inclusion of Mg2+ cations into the simulations causes perturbation of basepairing at the LE/HeIV junction, which does not, however, affect the protein binding. The rRNA-protein complex is mediated by a number of highly specific hydration sites with long-residing water molecules and two of them are bound throughout the entire 24-ns simulation. Long residing water molecules are seen also outside the RNA-protein contact areas with water-binding times substantially enhanced compared to simulations of free RNA. Long-residency hydration sites thus represent important elements of the three dimensional structure of rRNA. PMID- 15339801 TI - Physical detwinning of hemihedrally twinned hexagonal crystals of bacteriorhodopsin. AB - Hexagonal crystals of the membrane protein bacteriorhodopsin of space group P6 3 grown in lipidic cubic phase are twinned hemihedrally. It was shown that slow changes of salt concentration in the mother liquor lead to a split of crystals so that the split parts preserved high diffraction quality. Analysis of diffraction data from split crystals by Yeates statistic and Britton plot showed that the split parts are free of twinning. It is concluded that crystals of bacteriorhodopsin are composed of several macroscopic twinning domains with sizes comparable to the original crystal. The appearance of twinning domains during crystal growth and the mechanism of splitting are discussed. PMID- 15339802 TI - An automatic method for predicting transmembrane protein structures using cryo-EM and evolutionary data. AB - The transmembrane (TM) domains of many integral membrane proteins are composed of alpha-helix bundles. Structure determination at high resolution (<4 A) of TM domains is still exceedingly difficult experimentally. Hence, some TM-protein structures have only been solved at intermediate (5-10 A) or low (>10 A) resolutions using, for example, cryo-electron microscopy (cryo-EM). These structures reveal the packing arrangement of the TM domain, but cannot be used to determine the positions of individual amino acids. The observation that typically, the lipid-exposed faces of TM proteins are evolutionarily more variable and less charged than their core provides a simple rule for orienting their constituent helices. Based on this rule, we developed score functions and automated methods for orienting TM helices, for which locations and tilt angles have been determined using, e.g., cryo-EM data. The method was parameterized with the aim of retrieving the native structure of bacteriorhodopsin among near- and far-from-native templates. It was then tested on proteins that differ from bacteriorhodopsin in their sequences, architectures, and functions, such as the acetylcholine receptor and rhodopsin. The predicted structures were within 1.5 3.5 A from the native state in all cases. We conclude that the computational method can be used in conjunction with cryo-EM data to obtain approximate model structures of TM domains of proteins for which a sufficiently heterogeneous set of homologs is available. We also show that in those proteins in which relatively short loops connect neighboring helices, the scoring functions can discriminate between near- and far-from-native conformations even without the constraints imposed on helix locations and tilt angles that are derived from cryo-EM. PMID- 15339803 TI - Near-field scanning fluorescence microscopy study of ion channel clusters in cardiac myocyte membranes. AB - Near-field scanning optical microscopy (NSOM) has been used to study the nanoscale distribution of voltage-gated L-type Ca2+ ion channels, which play an important role in cardiac function. NSOM fluorescence imaging of immunostained cardiac myocytes (H9C2 cells) demonstrates that the ion channel is localized in small clusters with an average diameter of 100 nm. The clusters are randomly distributed throughout the cell membrane, with some larger fluorescent patches that high-resolution images show to consist of many small closely-spaced clusters. We have imaged unstained cells to assess the contribution of topography induced artifacts and find that the topography-induced signal is <10% of the NSOM fluorescence intensity. We have also examined the dependence of the NSOM signal intensity on the tip-sample separation to assess the contributions from fluorophores that are significantly below the cell surface. This indicates that chromophores > approximately 200 nm below the probe will have negligible contributions to the observed signal. The ability to quantitatively measure small clusters of ion channels will facilitate future studies that examine changes in protein localization in stimulated cells and during cardiac development. Our work illustrates the potential of NSOM for studying membrane domains and protein localization/colocalization on a length scale which exceeds that available with optical microscopy. PMID- 15339804 TI - Calculation of cyclodextrin binding affinities: energy, entropy, and implications for drug design. AB - The second generation Mining Minima method yields binding affinities accurate to within 0.8 kcal/mol for the associations of alpha-, beta-, and gamma-cyclodextrin with benzene, resorcinol, flurbiprofen, naproxen, and nabumetone. These calculations require hours to a day on a commodity computer. The calculations also indicate that the changes in configurational entropy upon binding oppose association by as much as 24 kcal/mol and result primarily from a narrowing of energy wells in the bound versus the free state, rather than from a drop in the number of distinct low-energy conformations on binding. Also, the configurational entropy is found to vary substantially among the bound conformations of a given cyclodextrin-guest complex. This result suggests that the configurational entropy must be accounted for to reliably rank docked conformations in both host-guest and ligand-protein complexes. In close analogy with the common experimental observation of entropy-enthalpy compensation, the computed entropy changes show a near-linear relationship with the changes in mean potential plus solvation energy. PMID- 15339805 TI - Structure and transport mechanism of the bacterial oxalate transporter OxlT. AB - Membrane proteins that belong to the major facilitator superfamily (MFS) are found in organisms across the evolutionary spectrum and mediate the transport of a variety of substrates ranging from small metabolites to neurotransmitters. The oxalate transporter (OxlT) is a representative MFS protein, and exchanges formate for oxalate across the cytoplasmic membrane of the organism Oxalobacter formigenes. Here, we present a structural model for the protein conformational changes that occur during oxalate transport by combining a three-dimensional map of the oxalate-bound, "closed" state of OxlT at 6.5 A determined by cryo-electron microscopy with a model of the "open" state of OxlT based on the atomic structures of the related transporters, glycerol-3-phosphate transporter (GlpT) and lactose permease (LacY). We demonstrate that the principal structural change associated with substrate transport is a concerted rocking movement of the two structurally similar halves of the protein relative to each other. Our structural model places two positively charged residues, Arg-272 and Lys-355 in the central cavity, suggesting that electrostatic interactions between these residues and the oxalate anion is a key step in generating the conformational change between the open and closed states of the transporter. PMID- 15339806 TI - QM/MM study of energy storage and molecular rearrangements due to the primary event in vision. AB - The energy storage and the molecular rearrangements due to the primary photochemical event in rhodopsin are investigated by using quantum mechanics/molecular mechanics hybrid methods in conjunction with high-resolution structural data of bovine visual rhodopsin. The analysis of the reactant and product molecular structures reveals the energy storage mechanism as determined by the detailed molecular rearrangements of the retinyl chromophore, including rotation of the (C11-C12) dihedral angle from -11 degrees in the 11-cis isomer to -161 degrees in the all-trans product, where the preferential sense of rotation is determined by the steric interactions between Ala-117 and the polyene chain at the C13 position, torsion of the polyene chain due to steric constraints in the binding pocket, and stretching of the salt bridge between the protonated Schiff base and the Glu-113 counterion by reorientation of the polarized bonds that localize the net positive charge at the Schiff-base linkage. The energy storage, computed at the ONIOM electronic-embedding approach (B3LYP/6-31G*:AMBER) level of theory and the S0-->S1 electronic-excitation energies for the dark and product states, obtained at the ONIOM electronic-embedding approach (TD-B3LYP/6 31G*//B3LYP/6-31G*:AMBER) level of theory, are in very good agreement with experimental data. These results are particularly relevant to the development of a first-principles understanding of the structure-function relations in prototypical G-protein-coupled receptors. PMID- 15339807 TI - Mesoscopic lateral diffusion in lipid bilayers. AB - The lateral diffusion in bilayers is modeled with a multiscale mesoscopic simulation. The methodology consists of two simulations, where the first employs atomistic models to obtain exact results for the mesoscopic model. The second simulation takes the results obtained from the first to parameterize an effective force field that is employed in a new coarse-grained model. The multiscale aspect of this scheme occurs at the point where the microscopic time-averaged results of the first simulation are employed to parameterize the second simulation that operates in a higher spatial and temporal domain. The results of both simulation schemes give quantitative information on the details associated with lipid lateral diffusion. PMID- 15339808 TI - A study of low pH-induced refolding of Env of avian sarcoma and leukosis virus into a six-helix bundle. AB - The fusion protein of avian sarcoma and leukosis virus is likely to fold into a six-helix bundle as part of its final configuration. A peptide, R99, inhibits fusion, probably by binding into the grooves of the triple-stranded coiled coil that becomes the central core of the six-helix bundle. The stages at which the envelope protein (Env) of avian sarcoma and leukosis virus subgroup A folds into a bundle during low pH-induced fusion were determined. Effector cells expressing Env were bound to target cells expressing the cognate receptor Tva, and intermediates of fusion were created. R99 was added and the extent of fusion inhibition was used to distinguish between a prebundle state with exposed grooves and a state in which the grooves were no longer exposed. The native conformation of Env was not sensitive to R99. But adding a soluble form of Tva to effector cells conferred sensitivity. Acidic pH applied at low temperature created an intermediate state of local hemifusion. Surprisingly, R99 caused these locally hemifused membranes to separate. This indicates that the grooves of Env were still exposed, that prebundle configurations of Env stabilized hemifused states, and that binding of R99 altered the conformation of Env. In the presence of an inhibitory lipid that blocks fusion before hemifusion, applying low pH at 37 degrees C created an intermediate in which R99 was without effect. This suggests that the six-helix bundle can form before hemifusion and that subsequent conformational changes, such as formation of the trimeric hairpin, are responsible for pore formation and/or growth. PMID- 15339809 TI - Defining the physical gate of a mechanosensitive channel, MscL, by engineering metal-binding sites. AB - The mechanosensitive channel of large conductance, MscL, of Escherichia coli is one of the best-studied mechanosensitive proteins. Although the structure of the closed or "nearly-closed" state of the Mycobacterium tuberculosis ortholog has been solved and mechanisms of gating have been proposed, the transition from the closed to the open states remains controversial. Here, we probe the relative position of specific residues predicted to line the pore of MscL in either the closed state or during the closed-to-open transition by engineering single-site histidine substitutions and assessing the ability of Ni2+, Cd2+ or Zn2+ ions to affect channel activity. All residues predicted to be within the pore led to a change in channel threshold pressure, although the direction and extent of this change were dependent upon the mutation and metal used. One of the MscL mutants, L19H, exhibited gating that was inhibited by Cd2+ but stimulated by Ni2+, suggesting that these metals bind to and influence different states of the channel. Together, the results derived from this study support the hypotheses that the crystal structure depicts a "nearly closed" rather than a "fully closed" state of MscL, and that a clockwise rotation of transmembrane domain 1 occurs early in the gating process. PMID- 15339810 TI - Negatively charged residues in the N-terminal of the AID helix confer slow voltage dependent inactivation gating to CaV1.2. AB - The E462R mutation in the fifth position of the AID (alpha1 subunit interaction domain) region in the I-II linker is known to significantly accelerate voltage dependent inactivation (VDI) kinetics of the L-type CaV1.2 channel, suggesting that the AID region could participate in a hinged-lid type inactivation mechanism in these channels. The recently solved crystal structures of the AID-CaVbeta regions in L-type CaV1.1 and CaV1.2 channels have shown that in addition to E462, positions occupied by Q458, Q459, E461, K465, L468, D469, and T472 in the rabbit CaV1.2 channel could also potentially contribute to a hinged-lid type mechanism. A mutational analysis of these residues shows that Q458A, Q459A, K465N, L468R, D469A, and T472D did not significantly alter VDI gating. In contrast, mutations of the negatively charged E461, E462, and D463 to neutral or positively charged residues increased VDI gating, suggesting that the cluster of negatively charged residues in the N-terminal end of the AID helix could account for the slower VDI kinetics of CaV1.2. A mutational analysis at position 462 (R, K, A, G, D, N, Q) further confirmed that E462R yielded faster VDI kinetics at +10 mV than any other residue with E462R >> E462K approximately E462A > E462N > wild-type approximately E462Q approximately E462G > E462D (from the fastest to the slowest). E462R was also found to increase the VDI gating of the slow CEEE chimera that includes the I-II linker from CaV1.2 into a CaV2.3 background. The fast VDI kinetics of the CaV1.2 E462R and the CEEE + E462R mutants were abolished by the CaVbeta2a subunit and reinstated when using the nonpalmitoylated form of CaVbeta2a C3S + C4S (CaVbeta2a CS), confirming that CaVbeta2a and E462R modulate VDI through a common pathway, albeit in opposite directions. Altogether, these results highlight the unique role of E461, E462, and D463 in the I-II linker in the VDI gating of high voltage activated CaV1.2 channels. PMID- 15339811 TI - Analysis of side-chain rotamers in transmembrane proteins. AB - We measured the frequency of side-chain rotamers in 14 alpha-helical and 16 beta barrel membrane protein structures and found that the membrane environment considerably perturbs the rotamer frequencies compared to soluble proteins. Although there are limited experimental data, we found statistically significant changes in rotamer preferences depending on the residue environment. Rotamer distributions were influenced by whether the residues were lipid or protein facing, and whether the residues were found near the N- or C-terminus. Hydrogen bonding interactions with the helical backbone perturbs the rotamer populations of Ser and His. Trp and Tyr favor side-chain conformations that allow their side chains to extend their polar atoms out of the membrane core, thereby aligning the side-chain polarity gradient with the polarity gradient of the membrane. Our results demonstrate how the membrane environment influences protein structures, providing information that will be useful in the structure prediction and design of transmembrane proteins. PMID- 15339812 TI - Computational analysis of DNA gyrase action. AB - DNA gyrase introduces negative supercoiling into circular DNA by catalyzing the passage of one DNA segment through another. The efficiency of the reaction is many times higher than that of other topological transformations. We analyze, by a computer simulation, the reaction selectivity for a model of DNA gyrase action that assumes existence of a free loop between the G- and T- DNA segments participating in the reaction. A popular model of this type assumed that the selectivity can be provided by the conformation of the DNA segment wrapped around the enzyme into the right-handed helix turn (G-segment). We simulated the distribution of the reaction products for this model. Equilibrium sets of DNA conformations with one segment of the double helix wrapped around the enzyme were constructed. From these sets we selected conformations that had a second segment properly juxtaposed with the first one. Assuming that the juxtapositions result in the strand-passing reaction, we calculated the reaction products for all these conformations. The results show that different products have to be formed if the enzyme acts according to the model. This conclusion can be extended for any model with a free loop between the G- and T-segments. An alternative model that is consistent with the major experimental observations and the computational analysis, is suggested. PMID- 15339813 TI - The protonation status of compound II in myoglobin, studied by a combination of experimental data and quantum chemical calculations: quantum refinement. AB - Treatment of met-myoglobin (FeIII) with H2O2 gives rise to ferryl myoglobin, which is closely related to compound II in peroxidases. Experimental studies have given conflicting results for this species. In particular, crystallographic and extended x-ray absorption fine-structure data have shown either a short (approximately 170 pm) or a longer (approximately 190 pm) Fe-O bond, indicating either a double or a single bond. We here present a combined experimental and theoretical investigation of this species. In particular, we use quantum refinement to re-refine a crystal structure with a long bond, using 12 possible states of the active site. The states differ in the formal oxidation state of the iron ion and in the protonation of the oxygen ligand (O2-, OH-, or H2O) and the distal histidine residue (with a proton on Ndelta1, Nepsilon2, or on both atoms). Quantum refinement is essentially standard crystallographic refinement, where the molecular-mechanics potential, normally used to supplement the experimental data, is replaced by a quantum chemical calculation. Thereby, we obtain an accurate description of the active site in all the different protonation and oxidation states, and we can determine which of the 12 structures fit the experimental data best by comparing the crystallographic R-factors, electron-density maps, strain energies, and deviation from the ideal structure. The results indicate that FeIII OH- and FeIV OH- fit the experimental data almost equally well. These two states are appreciably better than the standard model of compound II, FeIV O2-. Combined with the available spectroscopic data, this indicates that compound II in myoglobin is protonated and is best described as FeIV OH-. It accepts a hydrogen bond from the distal His, which may be protonated at low pH. PMID- 15339814 TI - Topographical pattern dynamics in passive adhesion of cell membranes. AB - Strong adhesion of highly active cells often nucleates focal adhesions, synapses, and related structures. Red cells lack such complex adhesion systems and are also nonmotile, but they are shown here to dynamically evolve complex spatial patterns beyond an electrostatic threshold for strong adhesion. Spreading of the cell onto a dense, homogeneous poly-L-lysine surface appears complete in <1 s with occasional blisters that form and dissipate on a similar timescale; distinct rippled or stippled patterns in fluorescently labeled membrane components emerge later, however, on timescales more typical of long-range lipid diffusion (approximately minutes). Within the contact zone, the anionic fluorescent lipid fluorescein phosphoethanolamine is seen to rearrange, forming worm-like rippled or stippled domains of <500 nm that prove independent of whether the cell is intact and sustaining a tension or ruptured. Lipid patterns are accompanied by visible perturbations in Band 3 distribution and weaker perturbations in membrane skeleton actin. Pressing down on the membrane quenches the lipid patterns, revealing a clear topographical basis for pattern formation. Counterion screening and membrane fluctuations likely contribute, but the results primarily highlight the fact that even in adhesion of a passive red cell, regions of strong contact slowly evolve to become interspersed with regions where the membrane is more distant from the surface. PMID- 15339815 TI - Dynamics of the DNA duplex formation studied by single molecule force measurements. AB - DNA is partly denatured in vitro by applying a force that mechanically separates the two strands of the double helix. Sudden reduction of the imposed displacement triggers spontaneous reannealing of the molecule. The corresponding force signals are measured by optical trapping interferometry for backward steps of various amplitudes and base sequence intervals. The measured signals frequently show plateaus of varying duration at discrete values that are dependent on the base sequence. Additional measurements are performed with proteins bound to the double helix. When the opening fork encounters such a protein during mechanical unzipping, force increases until the protein is ejected. This ejection induces fast release of tension and fast unzipping. Comparing our different measurements, we find that both DNA unzipping and the relaxation of tension in DNA are faster than the formation of the double helix. PMID- 15339816 TI - Equilibrium structure and folding of a helix-forming peptide: circular dichroism measurements and replica-exchange molecular dynamics simulations. AB - We have performed experimental measurements and computer simulations of the equilibrium structure and folding of a 21-residue alpha-helical heteropeptide. Far ultraviolet circular dichroism spectroscopy is used to identify the presence of helical structure and to measure the thermal unfolding curve. The observed melting temperature is 296 K, with a folding enthalpy of -11.6 kcal/mol and entropy of -39.6 cal/(mol K). Our simulations involve 45 ns of replica-exchange molecular dynamics of the peptide, using eight replicas at temperatures between 280 and 450 K, and the program CHARMM with a continuum solvent model. In a 30-ns simulation started from a helical structure, conformational equilibrium at all temperatures was reached after 15 ns. This simulation was used to calculate the peptide melting curve, predicting a folding transition with a melting temperature in the 330-350 K range, enthalpy change of -10 kcal/mol, and entropy change of 30 cal/(mol K). The simulation results were also used to analyze the peptide structural fluctuations and the free-energy surface of helix unfolding. In a separate 15-ns replica-exchange molecular dynamics simulation started from the extended structure, the helical conformation was first attained after approximately 2.8 ns, and equilibrium was reached after 10 ns of simulation. These results showed a sequential folding process with a systematic increase in the number of hydrogen bonds until the helical state is reached, and confirmed that the alpha-helical state is the global free-energy minimum for the peptide at low temperatures. PMID- 15339817 TI - Electroosmosis in transdermal iontophoresis: implications for noninvasive and calibration-free glucose monitoring. AB - Reverse iontophoresis uses a small low electric current to noninvasively extract blood analytes, e.g., glucose, across the skin. The simultaneous quantification of the analyte extracted and of an additional endogenous substance of fixed and known concentration in the body permits the blood level of interest to be found without the need for an invasive calibration procedure. The transport phenomena underlying this approach, applied to glucose monitoring, has been investigated in vitro, using Na+ and neutral model solutes as endogenous "internal standards" (specifically, urea, glycerol, mannitol, and sucrose). The cathodal extracted fluxes of glucose under conditions of modified skin permselectivity were related to those of the different, potential internal standards. Flux ratios depended upon the iontophoretic conditions and the size of the neutral internal standards, whereas high variability was observed with Na+. Constant flux ratios were obtained with mannitol, glycerol, urea, and sucrose for which the mechanism of electrotransport was identical to that of glucose. The advantage of using a neutral internal standard, however, must be weighed against the need to identify and validate the marker under physiological conditions and the additional analytical chemistry necessary for the practical quantification of this substance. PMID- 15339818 TI - Anomalous subdiffusion is a measure for cytoplasmic crowding in living cells. AB - Macromolecular crowding dramatically affects cellular processes such as protein folding and assembly, regulation of metabolic pathways, and condensation of DNA. Despite increased attention, we still lack a definition for how crowded a heterogeneous environment is at the molecular scale and how this manifests in basic physical phenomena like diffusion. Here, we show by means of fluorescence correlation spectroscopy and computer simulations that crowding manifests itself through the emergence of anomalous subdiffusion of cytoplasmic macromolecules. In other words, the mean square displacement of a protein will grow less than linear in time and the degree of this anomality depends on the size and conformation of the traced particle and on the total protein concentration of the solution. We therefore propose that the anomality of the diffusion can be used as a quantifiable measure for the crowdedness of the cytoplasm at the molecular scale. PMID- 15339819 TI - Determination of proton flux and conductance at pH 6.8 through single FO sectors from Escherichia coli. AB - We have developed a mathematical model in concert with an assay that allows us to calculate proton (H+) flux and conductance through a single FO of the F1FO ATP synthase. Lipid vesicles reconstituted with just a few functional FO from Escherichia coli were loaded with 250 mM K+ and suspended in a low K+ solution. The pH of the weakly buffered external solution was recorded during sequential treatment with the potassium ionophore valinomycin, the protonophore carbonyl cyanide 3-chlorophenylhydrazone, and HCl. From these pH traces and separate determinations of vesicle size and lipid concentration we calculate the proton conductance through a single FO sector. This methodology is sensitive enough to detect small (15%) conductance changes. We find that wild-type FO has a proton flux of 3100 +/- 500 H+/s/FO at a transmembrane potential of 106 mV (25 degrees C and pH 6.8). This corresponds to a proton conductance of 4.4 fS. PMID- 15339820 TI - The survival of mental health services: a pressing research agenda? PMID- 15339821 TI - Migraine: another headache for psychiatrists? PMID- 15339822 TI - Treating unhappiness--society needs palliative psychopharmacology. PMID- 15339823 TI - Stimulant psychosis: systematic review. AB - BACKGROUND: Psychosis associated with stimulant use is an increasing problem, but there is little research evidence about the nature of the problem and its management. AIMS: To critically review the literature on stimulant psychosis and sensitisation. METHOD: Systematic review of studies that have investigated stimulant use and psychosis in humans. The main outcome measures were increases in psychosis with stimulant use, and differences between stimulant users and non users. RESULTS: Fifty-four studies met the inclusion criteria. Experimental studies show that a single dose of a stimulant drug can produce a brief increase in psychosis ratings (a "response") in 50-70% of participants with schizophrenia and pre-existing acute psychotic symptoms, unaffected by the presence of antipsychotic medication. Those with schizophrenia who do not have acute psychotic symptoms respond, but less frequently (30%). There has been little research into the longer-term effects of use. CONCLUSIONS: Compliance with antipsychotic medication by someone with schizophrenia will not prevent a relapse or worsening of psychotic symptoms if stimulants are used. Low-dose antipsychotic treatment may be beneficial in stimulant users, to prevent sensitisation. PMID- 15339824 TI - Changes in distributed neural circuitry function in patients with first-episode schizophrenia. AB - BACKGROUND: A number of functional brain abnormalities have been reported in schizophrenia, but it remains to be determined which of them represent trait and state markers of the illness. AIMS: To delineate regional brain dysfunctions that remain stable and those that fluctuate during the course of schizophrenia. METHOD: A cohort of patients with first-episode schizophrenia and a matched group of control participants underwent functional magnetic resonance imaging on two occasions 6-8 weeks apart during performance of a working memory task. The patients' disease was in partial remission at the second scan. RESULTS: Relative to control participants, the function of the left dorsolateral prefrontal cortex, left thalamus and right cerebellum remained disturbed in the people with schizophrenia, whereas the dysfunction of the right dorsolateral prefrontal cortex, right thalamus, left cerebellum and cingulate gyrus normalised, with significant reduction in symptoms. CONCLUSIONS: These results suggest that dysfunction of the left fronto-thalamo-cerebellar circuitry is a relatively stable characteristic of schizophrenia, whereas disturbance of the right circuitry and cingulate gyrusis predominantly a state-related phenomenon. PMID- 15339825 TI - Age at onset and cognitive functioning in schizophrenia. AB - BACKGROUND: Impairments in cognitive functioning are common in schizophrenia, and the degree of impairment may be associated with the individual's age at onset of the disorder. AIMS: To examine the effect of age at onset on cognitive functioning using the California Verbal Learning Test, sub-tests from the Wechsler Memory Scale-Revised and sub-scales from the Wechsler Adult Intelligence Scale-Revised among families with schizophrenia. METHOD: The effect of age at onset on cognitive function in 237 people with schizophrenia from a population based sample was examined using linear mixed effects models with family as the random effect, and age, gender, chronicity of the illness and number of affected first-degree relatives as fixed effects. RESULTS: Impairment in verbal learning and memory was associated with earlier disease onset. No association was found for working memory or IQ. CONCLUSIONS: In patients with early-onset schizophrenia, verbal memory functions in particular should be taken into account in neuropsychological evaluation and efforts at remediation. PMID- 15339826 TI - Psychosis, victimisation and childhood disadvantage: evidence from the second British National Survey of Psychiatric Morbidity. AB - BACKGROUND: Adverse early circumstances may be more common in people who later develop psychotic disorders. AIMS: To use data from the second British National Survey of Psychiatric Morbidity to examine associations between psychotic disorders and a number of early victimisation experiences. METHOD: Psychiatric disorders were identified through structured assessment of adults resident in private households in Britain (n=8580). Respondents were asked whether they had experienced selected events displayed on cards. RESULTS: Compared with respondents with other psychiatric disorders or with none, the prevalence of every experience bar one was significantly elevated in those with definite or probable psychosis. The largest odds ratio was for sexual abuse. Controlling for depressed mood somewhat reduced the odds ratios for the individual experiences. CONCLUSIONS: In people with psychosis, there is a marked excess of victimising experiences, many of which will have occurred during childhood. This is suggestive of a social contribution to aetiology. PMID- 15339827 TI - Area and individual circumstances and mood disorder prevalence. AB - BACKGROUND: Associations have been demonstrated between contextual (area level) factors and a range of physical health outcomes, but their relationship with mental health outcomes is less well understood. AIMS: To investigate the relative strength of association between individual and area-level demographic and socio economic factors and mood disorder prevalence in the UK. METHOD: Cross-sectional data from 19 687 participants from the European Prospective Investigation into Cancer and Nutrition in Norfolk. RESULTS: Area deprivation was associated with current (12-month) mood disorders after adjusting for individual-level socio economic status (OR for top v. bottom quartile of deprivation scores 1.29, 95% CI 1.1-1.5, P<0.001). However, this association was small relative to those observed for individual marital and employment status. Significant residual area-level variation in current mood disorders (representing 3.6% of total variation, P=0.04) was largely accounted for by individual-level factors. CONCLUSIONS: The magnitude of the association between socio-economic status and mood disorders is greater at the individual level than at the area level. PMID- 15339828 TI - Ethnicity, social deprivation and psychological distress in adolescents: school based epidemiological study in east London. AB - BACKGROUND: In adults the prevalence of psychological distress varies in different ethnic groups, and this has been explained by differences in socio economic status. Is this also the case in adolescents? AIMS: To examine whether ethnic differences in prevalence of psychological distress in adolescents are associated with social deprivation. METHOD: A cross-sectional questionnaire survey was used to assess 2790 male and female pupils, aged 11-14 years, from a representative sample of 28 east London secondary schools. RESULTS: Rates of psychological distress were similar to rates in UK national samples in boys and girls. Bangladeshi pupils, although highly socially disadvantaged, had a lower risk of psychological distress (OR=0.63, 95% CI 0.4-0.9). Non-UK White girls had higher rates of depressive symptoms (OR=1.54, 95% CI1.1-2.2). CONCLUSIONS: High rates of depressive symptoms in non-UK White girls may be related to recent migration. Low rates of psychological distress in Bangladeshi pupils in this sample relative to White pupils, despite socio-economic disadvantage, could be associated with cultural protective factors that require further investigation. PMID- 15339829 TI - Depression in men attending a rural general practice: factors associated with prevalence of depressive symptoms and diagnosis. AB - BACKGROUND: Doctors are less likely to diagnose depression in men than in women. Little research has been conducted to explore the underlying reasons for this in rural settings, or to compare primary care doctors' and male patients' ratings of perceived depression. AIMS: To identify symptomatic and socio-demographic correlates of depression in men attending a rural practice, and to compare and contrast general practitioners' and patients' assessments of depression. METHOD: All male patients of working age attending a rural general practice over a 12 month period were invited to participate. RESULTS: Men reporting recent "chest pain" or "feeling tired/little energy", expressing low job enjoyment or with a previous diagnosis of depression were more likely to be scored above threshold on the Hospital Anxiety and Depression Scale-Depression sub-scale. There was little agreement between the doctors and their male patients about the degree of perceived depression. CONCLUSIONS: Educational interventions aimed at addressing the diagnosis of depression in men should take greater account of factors within a particular social setting. PMID- 15339830 TI - Preventing crime by people with schizophrenic disorders: the role of psychiatric services. AB - BACKGROUND: Knowledge of when and how to implement treatments to prevent criminal offending among people with schizophrenia is urgently needed. AIMS: To identify opportunities for interventions to prevent offending among men with schizophrenic disorders by tracking their histories of offending and admissions to hospital. METHOD: We examined 232 men with schizophrenic disorders discharged from forensic and general psychiatric hospitals. Data were collected from participants, family members and official records. RESULTS: More than three-quarters (77.8%) of the forensic patients had previously been admitted to general psychiatric services; 24.3% of the general psychiatric patients had a criminal record. Offences had been committed by 39.8% of the forensic patients and 10.8% of the general psychiatric patients before their first admission to general psychiatry, and after their first admission these 59 patients committed 195 non-violent and 59 violent offences. Subsequently, 49 of them committed serious violent offences that led to forensic hospital admission. The offenders were distinguished by a pervasive and stable pattern of antisocial behaviour evident from at least mid adolescence. CONCLUSIONS: General psychiatry requires resources in order to prevent criminal offending among a subgroup of patients with schizophrenic disorders. PMID- 15339831 TI - What British psychiatrists read: questionnaire survey of journal usage among clinicians. AB - BACKGROUND: The role of journals in disseminating research to clinicians is increasingly debated. Current measures of esteem for journals (e.g. impact factors) may not indicate clinical penetration. AIMS: To assess the perceived importance of different mental health journals to psychiatrists' clinical practice and compare this with impact factors. METHOD: Random samples of psychiatrists providing child and adolescent, adults of working age and old age services chose up to ten journals read or consulted with regard to their clinical work, ranking the top three. For these journals, comparisons were made with impact factors and importance as outlets for UK psychiatry research. RESULTS: A total of 560 questionnaires were completed (47%). Two membership journals (the British Journal of Psychiatry and the BMJ) were most read and highest ranked. Associations between impact factors, clinicians' ratings and importance as outlets for psychiatry papers varied. CONCLUSIONS: The results could lead to reconsideration of the importance of some journals. Academic assessments of the status of journals should not be assumed to reflect their influence on clinicians. PMID- 15339832 TI - Cell density and cortical thickness in Heschl's gyrus in schizophrenia, major depression and bipolar disorder. AB - There is evidence that the superior temporal gyrus and Heschl's gyrus within it are implicated in schizophrenia. We investigated neuronal and glial cell density and cortical thickness within Heschl's gyrus, using the optical disector to estimate cell density within cortical layers 3 and 5 in tissue derived postmortem from people with diagnoses of major depressive disorder, schizophrenia and bipolar disorder, compared with normal controls (n=15 per group). No significant difference in neuronal or glial cell density or in cortical thickness was observed between the groups; our findings therefore provide no support for the presence of cellular pathology within Heschl's gyrus in schizophrenia. PMID- 15339833 TI - The continuing story of dhat syndrome. PMID- 15339835 TI - Disability after trauma. PMID- 15339836 TI - Analysis of psychiatric in-patient violence. PMID- 15339837 TI - Analysing the efficacy of clozapine. PMID- 15339838 TI - Case reports still valuable. PMID- 15339839 TI - Focus on psychiatry in Egypt. PMID- 15339840 TI - Distribution of dehydration rates generated by maximal Gardos-channel activation in normal and sickle red blood cells. AB - The Ca(2+)-activated K+ channels of human red blood cells (RBCs) (Gardos channels, hIK1, hSK4) can mediate rapid cell dehydration, of particular relevance to the pathophysiology of sickle cell disease. Previous investigations gave widely discrepant estimates of the number of Gardos channels per RBC, from as few as 1 to 3 to as many as 300, with large cell-to-cell differences, suggesting that RBCs could differ extensively in their susceptibility to dehydration by elevated Ca2+. Here we investigated the distribution of dehydration rates induced by maximal and uniform Ca2+ loads in normal (AA) and sickle (SS) RBCs by measuring the time-dependent changes in osmotic fragility and RBC volume distributions. We found a remarkable conservation of osmotic lysis and volume distribution profiles during Ca(2+)-induced dehydration, indicating overall uniformity of dehydration rates among AA and SS RBCs. In light of these results, alternative interpretations were suggested for the previously proposed low estimates and heterogeneity of channel numbers per cell. The results support the view that stochastic Ca2+ permeabilization rather than Gardos-channel variation is the main determinant selecting which SS cells dehydrate through Gardos channels in each sickling episode. PMID- 15339841 TI - Macrovascular thrombosis is driven by tissue factor derived primarily from the blood vessel wall. AB - Leukocytes and leukocyte-derived microparticles contain low levels of tissue factor (TF) and incorporate into forming thrombi. Although this circulating pool of TF has been proposed to play a key role in thrombosis, its functional significance relative to that of vascular wall TF is poorly defined. We tested the hypothesis that leukocyte-derived TF contributes to thrombus formation in vivo. Compared to wild-type mice, mice with severe TF deficiency (ie, TF(-/-), hTF-Tg+, or "low-TF") demonstrated markedly impaired thrombus formation after carotid artery injury or inferior vena cava ligation. A bone marrow transplantation strategy was used to modulate levels of leukocyte-derived TF. Transplantation of low-TF marrow into wild-type mice did not suppress arterial or venous thrombus formation. Similarly, transplantation of wild-type marrow into low-TF mice did not accelerate thrombosis. In vitro analyses revealed that TF activity in the blood was very low and was markedly exceeded by that present in the vessel wall. Therefore, our results suggest that thrombus formation in the arterial and venous macrovasculature is driven primarily by TF derived from the blood vessel wall as opposed to leukocytes. PMID- 15339842 TI - Disruption of Plasmodium berghei merozoite surface protein 7 gene modulates parasite growth in vivo. AB - Merozoite invasion of red blood cells is crucial to the development of the parasite that causes malaria. Merozoite surface proteins (MSPs) mediate the first interaction between parasite and erythrocyte. In Plasmodium falciparum, they include a complex of products from at least 3 genes (msp1, msp6, and msp7), one of which, msp7, is part of a gene family containing 3 and 6 adjacent members in Plasmodium yoelii and Plasmodium falciparum, respectively. We have identified and disrupted msp7 in the Plasmodium berghei gene family. The protein is expressed in schizonts and colocalizes with MSP1. The synthesis and processing of MSP1 was unaffected in the parasite with the disrupted gene (MSP7ko). Disruption of msp7 was not lethal but affected blood-stage parasite growth. MSP7ko parasites initially grew more slowly than wild-type parasites. However, when reticulocytes were prevalent, the rate of increase in parasitemia was similar, suggesting that MSP7ko parasites prefer to invade and grow within reticulocytes. PMID- 15339843 TI - Phospholipase D1 regulates high-affinity IgE receptor-induced mast cell degranulation. AB - To investigate the role of phospholipase D (PLD) in FcepsilonRI signaling, the wild-type or the catalytically inactive forms of PLD1 or PLD2 were stably overexpressed in RBL-2H3 mast cells. FcepsilonRI stimulation resulted in the activation of both PLD1 and PLD2. However, PLD1 was the source of most of the receptor-induced PLD activity. There was enhanced FcepsilonRI-induced degranulation only in cells that overexpressed the catalytically inactive PLD1. This dominant-negative PLD1 enhanced FcepsilonRI-induced tyrosine phosphorylations of early signaling molecules such as the receptor subunits, Syk and phospholipase C-gamma which resulted in faster release of Ca(2+) from intracellular sources. Therefore, PLD1 negatively regulates signals upstream of the Ca(2+) response. However, FcepsilonRI-induced PLD activation required Syk and was downstream of the Ca(2+)response, suggesting that basal PLD1 activity rather than that activated by cell stimulation controlled these early signaling events. Dominant-negative PLD1 reduced the basal phosphatidic acid formation in unstimulated cells, which was accompanied by an increase in FcepsilonRI within the lipid rafts. These results indicate that constitutive basal PLD1 activity by regulating phosphatidic acid formation controls the early signals initiated by FcepsilonRI aggregation that lead to mast cell degranulation. PMID- 15339844 TI - Rod mutations associated with MYH9-related disorders disrupt nonmuscle myosin-IIA assembly. AB - MYH9-related disorders are autosomal dominant syndromes, variably affecting platelet formation, hearing, and kidney function, and result from mutations in the human nonmuscle myosin-IIA heavy chain gene. To understand the mechanisms by which mutations in the rod region disrupt nonmuscle myosin-IIA function, we examined the in vitro behavior of 4 common mutant forms of the rod (R1165C, D1424N, E1841K, and R1933Stop) compared with wild type. We used negative-stain electron microscopy to analyze paracrystal morphology, a model system for the assembly of individual myosin-II molecules into bipolar filaments. Wild-type tail fragments formed ordered paracrystal arrays, whereas mutants formed aberrant aggregates. In mixing experiments, the mutants act dominantly to interfere with the proper assembly of wild type. Using circular dichroism, we find that 2 mutants affect the alpha-helical coiled-coil structure of individual molecules, and 2 mutants disrupt the lateral associations among individual molecules necessary to form higher-order assemblies, helping explain the dominant effects of these mutants. These results demonstrate that the most common mutations in MYH9, lesions in the rod, cause defects in nonmuscle myosin-IIA assembly. Further, the application of these methods to biochemically characterize rod mutations could be extended to other myosins responsible for disease. PMID- 15339845 TI - Perturbed myelo/erythropoiesis in Lyn-deficient mice is similar to that in mice lacking the inhibitory phosphatases SHP-1 and SHIP-1. AB - The Lyn tyrosine kinase plays essential inhibitory signaling roles within hematopoietic cells by recruiting inhibitory phosphatases such as SH2-domain containing phosphatase-1 (SHP-1), SHP-2, and SH2-domain containing 5'-inositol phosphatase (SHIP-1) to the plasma membrane in response to specific stimuli. Lyn deficient mice display a collection of hematopoietic defects, including autoimmune disease as a result of autoantibody production, and perturbations in myelopoiesis that ultimately lead to splenomegaly and myeloid neoplasia. In this study, we demonstrate that loss of Lyn results in a stem/progenitor cell intrinsic defect leading to an age-dependent increase in myeloid, erythroid, and primitive hematopoietic progenitor numbers that is independent of autoimmune disease. Despite possessing increased numbers of erythroid progenitors, and a more robust expansion of these cells following phenylhydrazine challenge, Lyn deficient mice are more severely affected by the chemotherapeutic drug 5 fluorouracil, revealing a greater proportion of cycling progenitors. We also show that mice lacking SHIP-1 have defects in the erythroid and myeloid compartments similar to those in mice lacking Lyn or SHP-1, suggesting an intimate relationship between Lyn, SHP-1, and SHIP-1 in regulating hematopoiesis. PMID- 15339846 TI - Fas-ligand (CD178) and TRAIL synergistically induce apoptosis of CD40-activated chronic lymphocytic leukemia B cells. AB - Chronic lymphocytic leukemia (CLL) B cells become sensitive to Fas (CD95) mediated apoptosis 3 to 5 days after CD40 ligation. However, CD4+ cytotoxic T lymphocytes (CTLs) can kill CLL B cells via a Fas-ligand (CD178)-dependent process within 24 hours after CD40 cross-linking, when ligation of CD95 alone is insufficient to induce apoptosis. In addition to CD95, CD40-activated CLL cells also express DR5, a receptor for tumor-necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) that is expressed by CD4+ CTL. In addition, CD40 ligation in vitro and in vivo induces CLL cells to express the proapoptotic protein, BH3 interacting domain death agonist (Bid), which can facilitate crosstalk between mitochondrial-dependent, apoptosis-inducing pathways and death receptors, such as death receptor 5 (DR5). To evaluate whether ligation of CD95 and/or DR5 can induce apoptosis of CD40-activated CLL cells, we generated artificial cytotoxic effector cells that express both human TRAIL and CD178 (Chinese hamster ovary [CHO]-CD178/TRAIL) or only TRAIL (CHO-TRAIL) or CD178 (CHO-CD178). CHO CD178/TRAIL cells were significantly more effective in killing CD40-activated CLL cells than either CHO-TRAIL or CHO-CD178 and, unlike the latter, could kill CLL cells 24 hours after CD40 ligation. We conclude that CD40 ligation induces CLL cells to express the proapoptotic molecule Bid and the death receptors CD95 and DR5, the latter of which can act synergistically to induce caspase-dependent apoptosis of CD40-activated CLL B cells. PMID- 15339847 TI - Genetic variation in XPD predicts treatment outcome and risk of acute myeloid leukemia following chemotherapy. AB - The xeroderma pigmentosum group D (XPD) gene encodes a DNA helicase that functions in nucleotide excision repair of chemotherapy-induced DNA damage, the efficiency of which is predicted to be affected by a lysine to glutamine variant at codon 751. We hypothesized that this constitutive genetic variant may modify clinical response to chemotherapy, and we have examined its association with outcome following chemotherapy for acute myeloid leukemia (AML) in 341 elderly patients entered into the United Kingdom Medical Research Council AML 11 trial, and with the risk of developing chemotherapy-related AML. Among subjects treated for AML, disease-free survival at one year was 44% for lysine homozygotes, compared with 36% for heterozygotes and 16% for glutamine homozygotes (hazard ratio [HR], 1.30; 95% confidence interval [CI], 1.01-1.70; P = .04). Similarly, overall survival at one year was 38% for lysine homozygotes, 35% for heterozygotes, and 23% for glutamine homozygotes (HR, 1.18; 95% CI, 0.99-1.41; P = .07). Furthermore, homozygosity for the XPD codon 751 glutamine variant was associated with a significantly increased risk of developing AML after chemotherapy (odds ratio, 2.22 for Gln/Gln vs Lys/Lys; 95% CI, 1.04-4.74). These data suggest that the XPD codon 751 glutamine variant protects against myeloid cell death after chemotherapy. PMID- 15339848 TI - Recombinant human activated protein C reduces human endotoxin-induced pulmonary inflammation via inhibition of neutrophil chemotaxis. AB - Recombinant human activated protein C (rhAPC) is a natural anticoagulant with potentially important anti-inflammatory properties. In humans with severe sepsis, rhAPC treatment reduces mortality, but mechanisms responsible have not been well characterized. Accumulation of activated neutrophils in the lungs and other organs during severe infection contributes to sepsis-induced organ dysfunction, including acute inflammatory lung injury. Because neutrophils express an APC receptor, we hypothesized that immunomodulatory effects of rhAPC occur, in part, via modulation of neutrophil responses. To examine this issue, we performed a double-blinded, placebo-controlled study of rhAPC in a human model of endotoxin induced pulmonary inflammation. Administration of rhAPC significantly reduced leukocyte accumulation to the airspaces, independent of pulmonary cytokine or chemokine release. Neutrophils recovered from bronchoalveolar lavage fluid of volunteers receiving rhAPC demonstrated decreased chemotaxis ex vivo. Decreased neutrophil chemotaxis following exposure to rhAPC was confirmed in vitro. No differences were detected in gene expression, kinase activation, cytokine release, cell survival, or apoptosis of neutrophils recovered in the presence or absence of rhAPC. These studies demonstrate that rhAPC reduces both endotoxin induced accumulation of leukocytes in the airspaces and neutrophil chemotaxis. These rhAPC-induced effects on neutrophil function may represent a mechanism by which rhAPC improves survival in patients with sepsis. PMID- 15339849 TI - p53 gene deletion detected by fluorescence in situ hybridization is an adverse prognostic factor for patients with multiple myeloma following autologous stem cell transplantation. AB - We investigated the relevance of p53 deletions to the clinical outcome of patients with multiple myeloma (MM) treated with high-dose chemotherapy and autologous stem cell transplantation. Hemizygous p53 gene deletions were detected by fluorescence in situ hybridization in 10 of 105 (9.5%) patients studied. p53 deletions were associated with higher serum calcium (P = .0062) and creatinine (P = .013) levels, but there were no association with patient age, gender, beta2 microglobulin, C-reactive protein, hemoglobin, albumin or bone lytic lesions, or immunoglobulin isotype. There were no associations of p53 deletions with 13q deletions or translocations t(11;14) or t(4;14). Patients with p53 deletions had significantly shorter progression-free (median, 7.9 versus 25.7 months, P = .0324) and overall survival (median, 14.7 versus 48.1 months, P = .0008) than patients without a p53 deletion. A multivariate analysis confirmed p53 deletion was an independent prognostic factor predicting shortened progression-free (P = .0009) or overall survival (P = .0002) in patients with MM after high-dose chemotherapy and autologous stem cell transplantation. PMID- 15339850 TI - Possible roles for activating RAS mutations in the MGUS to MM transition and in the intramedullary to extramedullary transition in some plasma cell tumors. AB - To assess a possible role in tumor progression, the occurrence and type of K- and N-RAS mutations were determined in purified tumor cells, including samples from patients with premalignant monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM), and extramedullary plasma cell (PC) tumors (ExPCTs). Immunophenotypic aberrant PCs were flow sorted from 20 MGUS, 58 MM, and 13 ExPCT patients. One RAS mutation was identified in 20 MGUS tumors (5%), in contrast to a much higher prevalence of RAS mutations in all stages of MM (about 31%). Further, oncogene analyses showed that RAS mutations are not evenly distributed among different molecular subclasses of MM, with the prevalence being increased in MM-expressing cyclin D1 (P = .015) and decreased in MM with t(4;14) (P = .055). We conclude that RAS mutations often provide a genetic marker if not a causal event in the evolution of MGUS to MM. Surprisingly, RAS mutations were absent in bone marrow tumor cells from all patients with ExPCT, a result significantly different from intramedullary MM (P = .001). From 3 of 6 patients with paired intramedullary and extramedullary PCs and identical immunoglobulin heavy chain gene (IgH) sequences, RAS mutations were identified only in extramedullary PCs, suggesting a role for RAS mutations in the transition from intramedullary to extramedullary tumor. PMID- 15339851 TI - Platelet activation induces metalloproteinase-dependent GP VI cleavage to down regulate platelet reactivity to collagen. AB - Glycoprotein (GP) VI, the primary collagen receptor on platelets, has been shown to have variable expression, possibly as a consequence of immune modulation. The present study was designed to determine the mechanism by which GP VI clearance occurs. We found that direct activation of GP VI both by a GP VI-specific antibody and by GP VI ligands (collagen and convulxin) reduced binding of biotinylated convulxin to the stimulated platelets. Analysis of immunoblots of platelets and supernatants showed that the stimulated platelets contained less GP VI, while the soluble fraction contained a 57-kDa cleavage product. Stimulation of platelets with PAR-1 agonists (TRAP peptide and thrombin) also caused GP VI cleavage, although the amount of GP VI loss was less than that observed with direct GP VI ligands. The metalloproteinase (MMP) inhibitors GM6001 and TAPI prevented both the clearance of GP VI from the platelet surface and the appearance of the soluble cleavage product. Induction of GP VI cleavage caused specific down-regulation of collagen-induced platelet aggregation, providing a mechanism for the modulation of platelet responsiveness to this important platelet agonist. PMID- 15339852 TI - Combined effects of As4S4 and imatinib on chronic myeloid leukemia cells and BCR ABL oncoprotein. AB - Imatinib (STI571, Gleevec) is a tailored drug for chronic myelogenous leukemia (CML), whereas arsenic compounds were used as ancient remedies for CML with certain efficacy. The aim of this study was to investigate the potential benefit of combination therapy with imatinib and arsenic sulfide (As(4)S(4)). Analysis of cell proliferation and clonogenic ability showed that As(4)S(4) and imatinib exerted synergistic effects on both K562 cells and fresh CML cells. The effective concentrations on fresh CML cells were pharmacokinetically available in vivo but had much less inhibitory effect on CD34(+) cells from the nonleukemic donors. Examination of cell cycles showed that As(4)S(4) induced G(2)/M arrest whereas imatinib induced G(1) arrest. Using a number of parameters such as morphology, annexin V/propidium iodide (PI), mitochondrial transmembrane potential, caspase-3 activity, and Fas/Fas-L, the synergistic effects were revealed on induction of cell apoptosis, largely through the mitochondrial pathway. The 2 drugs also exhibited a synergistic effect in targeting BCR-ABL protein. While As(4)S(4) triggered its degradation and imatinib inhibited its tyrosine kinase activity, combined use of the 2 led to lower protein/enzymatic activity levels of BCR-ABL. Our in vitro data thus strongly suggest a potential clinical application of imatinib and As(4)S(4) combination on CML. PMID- 15339853 TI - Retinoids and myelomonocytic growth factors cooperatively activate RARA and induce human myeloid leukemia cell differentiation via MAP kinase pathways. AB - Use of all-trans-retinoic acid (ATRA) in combinatorial differentiation therapy of acute promyelocytic leukemia (APL) results in exceptional cure rates. However, potent cell differentiation effects of ATRA are so far largely restricted to this disease and long-term survival rates in non-APL acute myelogeneous leukemia (AML) remain unacceptably poor, requiring development of novel therapeutic strategies. We demonstrate here that myelomonocytic growth factors (granulocyte colony stimulating factor [G-CSF] and/or granulocyte macrophage colony-stimulating factor [GM-CSF]) potentiate differentiation effects of ATRA in different AML cell lines and primary cells from patients with myeloid leukemia. The ligand-dependent activities of endogenous and transiently expressed retinoic acid receptor alpha (RARalpha) isoforms can be potentiated by G/GM-CSF in U-937 cells and correlate with increased expression of ATRA-inducible RARalpha2 isoform. Specific inhibitors of mitogen mitogen-activated protein kinase (MAPK) (MEK)-1/-2 or p38 extracellular signal-related kinase (ERK) kinase diminish the ATRA as well as ATRA and G/GM-CSF-induced activation of the RARalpha proteins and decreased the differentiation-induced decline in cell numbers. Our data demonstrate that acting, at least in part, via the MAP kinase pathways, myelomonocytic growth factors enhance ATRA-dependent activation of the RARalpha isoforms and maturation of myeloid leukemia cells. These results suggest that combinatorial use of these agents may be effective in differentiation therapy of AML. PMID- 15339854 TI - Bacterial polysaccharide-protein conjugate vaccines. AB - Following demonstration that chemical conjugation of polysaccharide antigens to proteins could enhance their immunogenicity in the 1920s, interest in this approach to primary prevention of bacterial infections waned with the development and widespread use of antibiotics. Emergence of resistant bacteria rekindled interest in the late 20th century, which saw extremely rapid development and implementation of several vaccines which are already rapidly changing the epidemiology of childhood infections with Haemophilus influenzae type b, Streptococcus pneumoniae and Neisseria meningitidis. Others such as Group B streptococcus and Salmonella typhi infections may soon follow. However, several important questions about the immunology of these antigens remain unanswered and the long-term implications of reducing or eliminating the circulation of organisms which are more commonly nasopharyngeal commensals than pathogenic invaders are uncertain. PMID- 15339855 TI - Allogeneic blood transfusions: benefit, risks and clinical indications in countries with a low or high human development index. AB - The risks associated with allogeneic red blood cell (RBC) transfusions differ significantly between countries with low and high human development indexes (HDIs). In countries with a low HDI, the risk of infection (HIV, HBV, HCV and malaria) is elevated. In contrast, in countries with a high HDI, immunological reactions (haemolytic transfusion reactions, alloimmunization and immunosuppression) are predominant. Therefore the overall risk associated with RBC transfusions in low HDI countries is much more significant than that in high HDI countries. In view of these risks, the limited efficacy of RBC transfusion and its high costs, this procedure should be used sparingly and rationally. Therefore RBC transfusion protocols adapted to the local situation are essential. Such protocols should distinguish between physiological and haemoglobin-based transfusion triggers. In countries with a high HDI, relative tachycardia and hypotension, despite normovolaemia, ST-segment changes suggestive of myocardial ischaemia and an Hb level <6 g/dl can serve as general guidelines for transfusion. Higher haemoglobin transfusion triggers should be used for patients aged >80 years and those with coronary artery or cerebrovascular disease. In countries with a low HDI, clinical signs of circulatory failure or myocardial ischaemia and an Hb level <5 g/dl can serve as transfusion guidelines. PMID- 15339856 TI - Hepatitis vaccines. PMID- 15339857 TI - 5-HT7 receptor efficacy distribution throughout the canine stomach. AB - This study aimed to determine, quantify and explain regional differences in the relaxant response to the selective 5-HT(1) and 5-HT(7) receptor agonist 5 carboxamidotryptamine (5-CT) throughout the canine stomach. Longitudinal muscle strips from eight gastric corpus regions and six antrum regions were mounted for isotonic measurement. The 5-CT-induced relaxation was examined on a prostaglandin F(2alpha)-induced submaximal response, expressed as percentage of this response and fitted to the operational model of agonism (OMOA). 5-HT(7) receptor messenger RNA (mRNA) expression was compared by means of quantitative PCR. 5-CT inhibited PGF(2alpha)-induced tonic contraction (corpus) and increase of phasic contraction amplitude (antrum). The consistent antagonism produced by the selective 5-HT(7) receptor antagonist SB-269970 (10 nm, pA(2) estimates 8.2-8.9) confirmed that in every region, the inhibition by 5-CT was 5-HT(7) receptor mediated. However, variation in the maximum effect (61-108%) and pEC(50) (6.4-8.6) was observed throughout the different regions. The OMOA explained these differences as differences in the efficacy parameter tau (ratio of receptor density and coupling efficiency; log tau estimates ranging from 0.1 to 2.1). The log tau gradient decreases going from the lesser to the greater curvature. A proportional difference (68%) in the relative expression of 5-HT(7) receptor mRNA between the lesser and the greater curvature indicates that differences in receptor density contribute to the observed functional differences. This study illustrates that 5 HT(7) receptors are present throughout the ventral wall of the canine stomach, but the efficacy (expressed as log tau) is clearly greater close to the lesser curvature. Differences in 5-HT(7) receptor expression at least partially explain the functional differences. PMID- 15339858 TI - CB1 receptor agonist and heroin, but not cocaine, reinstate cannabinoid-seeking behaviour in the rat. AB - We recently provided evidence for a functional link between cannabinoid and opioid endogenous systems in relapse to heroin-seeking behaviour in rats. In the present study, we aimed at investigating whether the previously observed cross talk between cannabinoids and opioids could be extended to mechanisms underlying relapse to cannabinoid-seeking behaviour after a prolonged period of abstinence. In rats previously trained to intravenously self-administer the synthetic cannabinoid receptor (CB1) agonist WIN 55,212-2 (12.5 microg kg(-1) inf(-1)) under a fixed ratio (FR1) schedule of reinforcement, noncontingent nonreinforced intraperitoneal (i.p.) priming injections of the previously self-administered CB1 agonist (0.25 and 0.5 mg kg(-1)) as well as heroin (0.5 mg kg(-1)), but not cocaine (10 mg kg(-1)), effectively reinstate cannabinoid-seeking behaviour following 3 weeks of extinction. The selective CB1 receptor antagonist SR 141716A (0.3 mg kg(-1) i.p.) does not reinstate responding when given alone, but completely prevents the cannabinoid-seeking behaviour triggered by WIN 55,212-2 or heroin primings. The nonselective opioid antagonist naloxone (1 mg kg(-1) i.p.) has no effect on operant behaviour per se, but significantly blocks cannabinoid- and heroin-induced reinstatement of cannabinoid-seeking behaviour. These results provide the first evidence of drug-induced reinstatement of cannabinoid-seeking behaviour, and further strengthen previous findings on a cross-talk between the endogenous cannabinoid and opioid systems in relapse mechanisms to drug-seeking. PMID- 15339859 TI - Mechanisms intrinsic to 5-HT2B receptor-induced potentiation of NMDA receptor responses in frog motoneurones. AB - In the presence of NMDA receptor open-channel blockers [Mg(2+); (+)-5-methyl 10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801); 1-amino-3,5 dimethyladamantane (memantine)] and TTX, high concentrations (30-100 microm) of either 5-hydroxytryptamine (5-HT) or alpha-methyl-5-hydroxytryptamine (alpha-Me-5 HT) significantly potentiated NMDA-induced depolarizations of frog spinal cord motoneurones. Potentiation was blocked by LY-53,857 (10-30 microm), SB 206553 (10 microm), and SB 204741 (30 microm), but not by spiroxatrine (10 microm), WAY 100,635 (1-30 microm), ketanserin (10 microm), RS 102221 (10 microm), or RS 39604 (10-20 microm). Therefore, alpha-Me-5-HT's facilitatory effects appear to involve 5-HT(2B) receptors. These effects were G-protein dependent as they were prevented by prior treatment with guanylyl-5'-imidodiphosphate (GMP-PNP, 100 microm) and H Arg-Pro-Lys-Pro-Gln-Gln-D-Trp-Phe-D-Trp-D-Trp-Met-NH(2) (GP antagonist 2A, 3-6 microm), but not by pertussis toxin (PTX, 3-6 ng ml(-1), 48 h preincubation). This potentiation was not reduced by protein kinase C inhibition with staurosporine (2.0 microm), U73122 (10 microm) or N-(2-aminoethyl)-5 isoquinolinesulfonamide HCl (H9) (77 microm) or by intracellular Ca(2+) depletion with thapsigargin (0.1 microm) (which inhibits Ca(2+)/ATPase). Exposure of the spinal cord to the L-type Ca(2+) channel blockers nifedipine (10 microm), KN-62 (5 microm) or gallopamil (100 microm) eliminated alpha-Me-5-HT's effects. The calmodulin antagonist N-(6-aminohexyl)-5-chloro-1-naphtalenesulfonamide (W7) (100 microm) diminished the potentiation. However, the calcium/calmodulin-dependent protein kinase II (CaM Kinase II) blocker KN-93 (10 microm) did not block the 5 HT enhancement of the NMDA responses. In summary, activation of 5-HT(2B) receptors by alpha-Me-5-HT facilitates NMDA-depolarizations of frog motoneurones via a G-protein, a rise in [Ca(2+)](i) from the entry of extracellular Ca(2+) through L-type Ca(2+) channels, the binding of Ca(2+) to calmodulin and a lessening of the Mg(2+) -produced open-channel block of the NMDA receptor. PMID- 15339860 TI - Functional expression of the serotonin 5-HT7 receptor in human glioblastoma cell lines. AB - Serotonin 5-HT(7) receptors are present in astrocytes. Understanding their role in this type of cell would greatly benefit from the identification of astroglial cell lines expressing this receptor type. The aim of the present study was to assess the expression of native 5-HT(7) receptors and 5-HT(7) receptor mRNA in a number of human glioblastoma cell lines, by means of cAMP measurements, Western blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. 5-Hydroxytryptamine (5-HT), 5-carboxamidotryptamine (5-CT), 5 methoxytryptamine (5-MeOT) and 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) induced concentration-dependent stimulations of cAMP accumulation in the human glioblastoma cell lines, U-373 MG, U-138 MG, U-87 MG, DBTRG-05MG, T98G, H4, CCF STTG1 and Hs 683. The rank order of potency was 5-CT>5-HT=5-MeOT>>8-OH-DPAT. The effect of 5-CT was inhibited in a concentration-dependent manner by the selective 5-HT(7) receptor antagonist SB-269970 in all human glioblastoma cells. Schild analyses yielded slope factors close to unity (0.89-1.13) and pA(2) values of 8.69-9.05. Western blot analysis revealed the presence of immunoreactive bands corresponding to the human 5-HT(7) receptor in extracts of all human glioblastoma cell lines. The presence of the three splice variants of the 5-HT(7) receptor (5 HT(7(a/b/d))) was visualized by RT-PCR analysis with specific primers in all human glioblastoma cell lines. In conclusion, human glioblastoma cell lines express functional 5-HT(7) receptors and the three splice variants of the corresponding mRNA. These cell lines could serve as model systems of native 5 HT(7) receptors in glial cells to investigate their putative role in processes like release of neurotrophic factors or inflammatory cytokines. PMID- 15339861 TI - Alterations in glucose metabolism by cyclosporine in rat brain slices link to oxidative stress: interactions with mTOR inhibitors. AB - Co-administration of the calcineurin inhibitor cyclosporine and the mTOR inhibitors sirolimus or everolimus increases the efficacy of immunosuppression after organ transplantation. However, clinical studies showed enhancement of cyclosporine toxicity. To characterize the biochemical mechanisms involved, we assessed the time-dependent effects of cyclosporine in combination with mTOR inhibitors on energy production (ex vivo (31)P-MRS), glucose metabolism (ex vivo (13)C-MRS), and reactive oxygen species (ROS) formation (using the fluorescent agent 2',7'-dichlorofluorescein diacetate) in perfused rat brain slices. Cyclosporine alone inhibited energy production (ATP: 75+/-9%), the Krebs cycle (4 (13)C-glutamate from 1-(13)C-glucose: 61+/-27%), and oxidative phosphorylation (NAD(+): 62+/-25%) after 4 h of perfusion. After 10 h, activation of anaerobic glycolysis (3-(13)C-lactate: 140+/-17%) compensated for inhibition of mitochondrial energy production and lowered the intracellular pH. ROS formation was increased after 4 h (285+/-55% of untreated control), but not after 10 h. mTOR inhibitors alone inhibited lactate production. When combined with cyclosporine, sirolimus enhanced cyclosporine-induced inhibition of energy metabolism (ATP: 64+/-9%) and ROS formation (367+/-46%). Most importantly, sirolimus inhibited cytosolic glycolysis and therefore compensation for cyclosporine-induced ATP reduction after 10 h. In contrast to sirolimus, everolimus antagonized cyclosporine-induced inhibition of mitochondrial energy metabolism (ATP: 91+/-7%) and ROS formation (170+/-49%). The antioxidant tocopherol antagonized all cyclosporine effects on cell metabolism. Cyclosporine time-dependently inhibited mitochondrial metabolism and increased ROS, followed by compensation involving anaerobic glycolysis. Everolimus antagonized cyclosporine-induced mitochondrial dysfunction, whereas sirolimus inhibited compensatory anaerobic glycolysis, thus enhancing cyclosporine's negative effects. ROS play the key role in mediating the negative effects of cyclosporine on cell energy metabolism. PMID- 15339862 TI - Bolus injection of human UII in conscious rats evokes a biphasic haemodynamic response. AB - A biphasic cardiovascular response to bolus i.v. injection of human urotensin II (hUII, 3 nmol kg(-1)) in conscious, male, Sprague-Dawley (SD) rats was identified and underlying mechanisms were explored. Initially (0-5 min) there was tachycardia, hypotension and mesenteric and hindquarters vasodilatation; later (30-120 min), tachycardia, hindquarters vasodilatation and a modest rise in blood pressure occurred. Pretreatment with indomethacin or N(G) nitro-l-arginine methylester (l-NAME) reduced the mesenteric vasodilator response to hUII, and abolished the late tachycardia and hindquarters vasodilatation. Indomethacin also abolished the hypotension and early hindquarters vasodilatation, and substantially reduced the initial tachycardia. Indomethacin and l-NAME together prevented all haemodynamic responses to hUII. Inhibition of inducible NOS had no effect on responses to hUII, whereas inhibition of neuronal NOS reduced the delayed tachycardic response to hUII but did not significantly affect the vasodilatation. Only the initial tachycardic response to hUII was antagonised by propranolol. In spontaneously hypertensive rats (SHR), the initial haemodynamic responses to hUII were qualitatively similar to those in SD rats, although there was also a modest renal vasodilatation. The secondary response comprised a smaller tachycardia and a small rise in blood pressure, with no significant hindquarters vasodilatation. Haemodynamic responses to hUII were not enhanced by endothelin and angiotensin receptor antagonism in either SD rats or in SHRs. One interpretation of these results is that the primary response to bolus injection of hUII is prostanoid- or prostanoid- and NO-mediated (mesenteric vasodilatation) and that this triggers secondary events, which are dependent on eNOS (hindquarters vasodilatation) and neuronal NOS (tachycardia). PMID- 15339863 TI - Investigation of the effect of the farnesyl protein transferase inhibitor R115777 on isoprenylation and intracellular signalling by the prostacyclin receptor. AB - The human (h) and mouse (m) prostacyclin receptors (IPs) undergo isoprenylation through attachment of a C-15 farnesyl moiety within their conserved carboxyl terminal -CSLC sequences. Herein, the effects of a novel farnesyl transferase inhibitor R115777 on signalling by the hIP and mIP, overexpressed in human embryonic kidney 293 cells, and by the hIP endogenously expressed in human erythroleukaemia cells were investigated. R115777 significantly impaired IP mediated cyclic AMP generation (IC(50) 0.37-0.60 nm) and intracellular calcium ([Ca(2+)](i)) mobilization (IC(50) 37-65 nm), but had no effect on signalling by the control nonisoprenylated beta(2) adrenergic receptor or the alpha or beta isoforms of the human thromboxane A(2) receptor (TP). Additionally, R115777 significantly reduced IP-mediated cross-desensitization of signalling by the TP alpha, but not by the TP beta, isoform of the human TP and impaired the farnesylation-dependent processing of the chaperone HDJ-2 protein (IC(50) 4.5 nm). Furthermore, R115777 fully impaired isoprenylation of both the Ha-Ras(WT) and Ha-Ras(CSLC) in vitro and in whole cells confirming that, unlike N-Ras and Ki Ras, the -CSLC motif associated with the IP cannot support alternative geranylgeranylation in the presence of R115777 and does not act as a substrate for geranylgeranyl transferase 1 in vitro or in whole cells. In conclusion, these data confirm that R115777 potently impairs IP isoprenylation and signalling, and suggest that clinically it may not only target Ras proteins but may also disrupt IP isoprenylation, events which could impact on physiologic processes in which prostacyclin and its receptor are implicated. PMID- 15339864 TI - Cardiovascular activity of rasagiline, a selective and potent inhibitor of mitochondrial monoamine oxidase B: comparison with selegiline. AB - Selegiline is used for treating Parkinson's disease. Despite its efficacy, the clinical use of selegiline in combination with l-dihydroxphenylalanine in Parkinsonian patients is hampered by cardiovascular complications, such as hypotension. This study was designed to compare in rats the cardiovascular effects of selegiline and rasagiline, their metabolites l-methamphetamine and aminoindan (TVP-136), respectively, and the second rasagiline metabolite non monoamine oxidase (MAO) inhibitor TVP-1022 (N-propargyl-1S(-)aminoindan). Intravenous (i.v.) administration of selegiline and rasagiline (1 mg kg(-1)) to anaesthetized rats (thiobutabarbital, 100 mg kg(-1), i.p.) did not affect mean arterial pressure (MAP), carotid blood flow (CBF) or carotid vascular resistance (CVR). Selegiline (10 mg kg(-1), i.v.) decreased MAP, CBF and increased CVR. In contrast, rasagiline (10 mg kg(-1), i.v.) caused a small transient decrease in MAP, while CBF and CVR were unchanged. l-methamphetamine (1 mg kg(-1), i.v.) administration provoked a dramatic and long-lasting depressor response, decreased CBF and increased CVR. In contrast, injection of aminoindan or TVP-1022 at a similar dose produced gradual nonsignificant decreases in MAP and CBF. Chronic oral treatment (21 days) of awake rats with selegiline at 10 mg kg(-1) decreased systolic blood pressure (SBP), diastolic blood pressure (DBP), and MAP, whereas heart rate was unaffected. Since the effective MAO-B inhibitory and clinical dose of rasagiline is about one-tenth that of selegiline, administration of 1 mg kg( 1) day(-1) rasagiline resulted in moderate decreases in SBP, DBP, and MAP, which were significantly lower than those caused by the 10 mg kg(-1) day(-1) dose of selegiline. These findings indicate that rasagiline, when given at doses equivalent to selegiline, is less likely to be hypotensive. PMID- 15339865 TI - Woldemar Mobitz and His 1924 classification of second-degree atrioventricular block. AB - Woldemar Mobitz, an early 20th century German internist, analyzed arrhythmias by graphing the relationship of changing atrial rates and premature beats to AV conduction. Through an astute mathematical approach, he was able to classify second-degree atrioventricular block into 2 types, subsequently referred to as Mobitz type I (Wenckebach) and Mobitz type II (Hay). Type I AV block was most often due to digitalis and was reversible. There were no associated pathological findings. Type II AV block frequently progressed to complete AV block and was associated with seizures, death, and pathological findings. PMID- 15339866 TI - ACC/AHA 2004 guideline update for coronary artery bypass graft surgery: summary article: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee to Update the 1999 Guidelines for Coronary Artery Bypass Graft Surgery). PMID- 15339867 TI - Images in cardiovascular medicine. "Shrink-wrapped" permanent pacemaker. PMID- 15339868 TI - Images in cardiovascular medicine. Transplant vasculopathy: evaluation with multi detector computed tomography. PMID- 15339869 TI - ACC/AHA guidelines for the management of patients with ST-elevation myocardial infarction: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee to Revise the 1999 Guidelines for the Management of Patients with Acute Myocardial Infarction). PMID- 15339870 TI - An exploration of street drinking in Drumchapel, Scotland. AB - This paper describes the results of a survey undertaken to contribute to a health needs assessment of persistent street drinkers in Drumchapel, Greater Glasgow. Street drinkers, members of the public and local service providers [including agency professionals (community workers; health workers; social workers) and others (e.g. shopkeepers)] were interviewed. Street drinkers were generally male, single, unemployed and living in their own (rented) accommodation. Not being homeless, they gave volitional reasons for drinking in the street, centred on social aspects and the cost of drinking in traditional establishments/public houses. Half the group said they did not do all their drinking on the street, suggesting alternatives may be possible. Uptake of food offered to respondents being interviewed was low, and self-reported diet was poor in vitamins and unvaried. Awareness of services was low in the street drinking group, with friends/family remaining the most common source of support. Public concern was mainly with the visibility of the group and the negative impact this was perceived to have on the community; however, service providers focused on health risks associated with street drinking. Consensus on the need for some accommodation/centre for drinkers with access to services was established across all groups. PMID- 15339872 TI - Direct renin assay and plasma renin activity assay compared. PMID- 15339874 TI - Management of venous thromboembolism: present and future. PMID- 15339875 TI - Initial treatment of venous thromboembolism. AB - Adequate initial anticoagulant therapy of deep venous thrombosis (DVT) is required to prevent thrombus growth and pulmonary embolism (PE). Intravenous unfractionated heparin (UFH) is being replaced by low-molecular-weight heparin (LMWH) as the anticoagulant of choice for initial treatment of venous thromboembolism (VTE). Both agents are relatively safe and effective when used to treat VTE, with LMWH suitable for outpatient therapy because of improved bioavailability and more predictable anticoagulant response. Serious potential complications of heparin therapy, such as heparin-induced thrombocytopenia (HIT) and osteoporosis, seem less common with LMWH. The potential for fetal harm and changes in maternal physiology complicate the treatment of VTE during pregnancy. Although systemic thrombolysis is used in patients with massive PE and in some patients with proximal DVT, controversy persists with respect to appropriate patient selection for this intervention. PMID- 15339876 TI - Long-term management of patients after venous thromboembolism. AB - Long-term treatment of venous thromboembolism (VTE) focuses mainly on the duration of anticoagulant therapy, usually with vitamin K (VK) antagonists. The duration of therapy should be individualized based on the risk of recurrent VTE if treatment were stopped and the risk of bleeding if treatment were continued. The risk of recurrence is low if thrombosis was provoked by a major reversible risk factor such as surgery; 3 months of treatment is usually adequate for such patients. The risk of recurrence is high if thrombosis was unprovoked ("idiopathic") or associated with an irreversible risk factor such as cancer; anticoagulant treatment for at least 6 months, and often indefinitely, is indicated for such patients. Risk of recurrence is intermediate if thrombosis was associated with a minor transient risk factor; such patients can be treated for 3 to 6 months. Within each of these categories, presentation as pulmonary embolism, >1 previous VTE, an underlying malignancy, an antiphospholipid antibody, or selected hereditary thrombophilic states favor more prolonged therapy, whereas isolated distal deep vein thrombosis, high risk of bleeding, and patient preference favor shorter treatment. The optimal intensity of anticoagulant therapy with VK antagonists corresponds to a target international normalized ratio of 2.5 (range, 2.0 to 3.0). Long-term treatment with low-molecular-weight heparin is an alternative to VK-antagonist therapy and is usually preferable in patients with active cancer. Oral direct thrombin inhibitors also appear suitable for long-term prevention of recurrent VTE but await regulatory approval and comparison with VK antagonists. PMID- 15339877 TI - New anticoagulants for treatment of venous thromboembolism. AB - Treatment of venous thromboembolism (VTE) usually starts with concomitant administration of heparin or low-molecular-weight heparin (LMWH) and a vitamin K antagonist. The parenteral anticoagulant, which is given for at least 5 days, is stopped once the vitamin K antagonist produces a therapeutic level of anticoagulation. Although the introduction of LMWH has simplified the initial treatment of VTE, problems remain. LMWH must be given by daily subcutaneous (SC) injection and vitamin K antagonists require routine coagulation monitoring, which is inconvenient for patients and physicians. Recently, 3 new anticoagulants have been introduced in an attempt to overcome these limitations. These include fondaparinux and idraparinux, synthetic analogs of the pentasaccharide sequence that mediates the interaction of heparin and LMWH with antithrombin, and ximelagatran, an orally active inhibitor of thrombin. These agents produce a predictable anticoagulant response; thus, routine coagulation monitoring is unnecessary. Because they do not bind to platelets or platelet factor 4, fondaparinux and idraparinux do not cause heparin-induced thrombocytopenia (HIT). Unlike vitamin K antagonists, ximelagatran has a rapid onset of action, thereby obviating the need for concomitant administration of a parenteral anticoagulant when starting treatment. The lack of an antidote for these new agents is a drawback, particularly for idraparinux, which has a long half-life. PMID- 15339878 TI - Invasive approaches to treatment of venous thromboembolism. AB - Deep vein thrombosis (DVT) occurs in one-quarter of a million individuals annually in the United States and results in significant disability from pulmonary embolism and chronic venous insufficiency, especially when the proximal iliofemoral is involved. Treatment has centered on early institution of adequate anticoagulation to prevent thrombus propagation and embolism, but anticoagulation alone does not always restore venous patency and many patients are left with venous outflow obstruction and valvular incompetence-the anatomic underpinnings of the postthrombotic syndrome. Various strategies have been used to restore patency of thrombosed veins, including open surgical thrombectomy, pharmacological thrombolysis, and percutaneous mechanical thrombectomy. Each modality has benefits and shortcomings. Surgical thrombectomy had previously been abandoned secondary to poor long-term results. More recently, with improved techniques and better patient selection, surgical thrombectomy has regained a therapeutic role in treating acute DVT in young patients with short segment occlusions. The advent of percutaneous techniques has allowed thrombolysis, percutaneous mechanical thrombectomy, and stenting to be used in conjunction with each other-allowing for better resolution of venous clot burden than when an individual modality is used alone. Practitioners who treat patients with DVT should be familiar with all the options available to restore venous patency, preserve valvular function, and thereby minimize the risk of late postthrombotic complications. PMID- 15339879 TI - Interaction of endotoxins with Toll-like receptor 4 correlates with their endotoxic potential and may explain the proinflammatory effect of Brucella spp. LPS. AB - Endotoxins displaying differences in the chemical structure of their lipid A were used to induce the expression of chemokines in the human monocytic THP-1 cell line. LPS from two enterobacterial species such as Escherichia coli and Yersinia enterocolitica induced mRNA expression of IFN-gamma-inducible protein (IP)-10, macrophage-inflammatory protein (MIP)-1alpha, MIP-1beta, monocyte chemoattractant protein (MCP)-1 and IL-8. LPS from the non-enterobacterial genera Brucella and Ochrobactrum induced the expression of these chemokines to a lower extent. Attempts to address the signaling routes involved in these responses were carried out in transiently transfected HEK293 cells. Induction of kappaB-driven transcriptional activity by enterobacterial LPS was observed in cells transfected with TLR-4 alone, although co-transfection of TLR-4, MD-2 and CD14 provided optimal induction. The response to Brucella spp. and Ochrobactrum anthropi LPS was only significant at the concentration of 10 microg/ml. These data indicate that LPS from Brucella spp. and O. anthropi, which contain lipid A moieties with structural features different from those of Enterobacteriaceae elicit biochemical signaling via TLR-4 only at high concentrations. Neither TLR-1, TLR-2 and TLR-6 nor heterodimeric combinations of these receptor molecules are involved. Conversely, the ability of LPS to activate the TLR-4 route is a reliable molecular biomarker for endotoxicity. PMID- 15339880 TI - Effect of c-fos overexpression on development and proliferation of peritoneal B cells. AB - We examined effects of c-fos overexpression on the development and property of peritoneal B-1 cells using transgenic (H2-c-fos) mice carrying the c-fos gene under the control of the constitutive H-2Kb promoter. The number of B-1b cells in the peritoneal cavity of H2-c-fos mice was 4-fold larger than that in control littermates. Although the numbers of total peritoneal B cells and B-1a cells were similar between them, the peritoneal B-2 cell number in H2-c-fos mice was reduced to 50% of control littermates, suggesting the effect of c-fos overexpression on a balance of B-1b and B-2 cells in a peritoneal cavity. Adoptive transfer experiments with hematopoietic stem cells of H2-c-fos and control mice into irradiated H2-c-fos mice demonstrated that the augmentation of B-1b cells is due to the c-fos effect in B cells and the effect on environment of the peritoneal cavity of H2-c-fos mice. When peritoneal B cells were cultured with LPS in the presence or absence of IL-4, cell proliferation of B-1b cells was the highest among these peritoneal B cell subsets, and the proliferation of H2-c-fos B-1b cells was 3-fold higher than that of control B-1b cells. This augmentation is due to the c-fos effect in B cells. IgG1 production of B-1b cells in these cultures was slightly higher than those of B-1a and peritoneal B-2 cells. Thus, the c-fos overexpression augments development of B-1b cells in a peritoneal cavity and proliferation of peritoneal B-1b cells to LPS. PMID- 15339881 TI - Auto-reactive B cells against peripheral antigen, desmoglein 3, escape from tolerance mechanism. AB - To examine the mechanism of B cell tolerance against natural peripheral self antigen, we generated transgenic mice expressing IgM specific for desmoglein 3 (Dsg3) from AK7 monoclonal antibody which itself does not induce blisters. Dsg3 is mainly expressed on stratified squamous epithelium and is the target antigen of an autoimmune bullous disease, pemphigus vulgaris. Transgenic B cells reactive to Dsg3 were observed in the spleen and lymph node. Although these B cells are autoreactive, they did not develop into B1 B cells. These B cells were functionally competent and anti-Dsg3 IgM was detected in the serum and on the keratinocyte cell surface. These results indicate that auto-reactive B cells against peripheral antigen (Dsg3) are able to develop in the presence of Dsg3 but are ignored by the immune system. PMID- 15339882 TI - Genetic basis of tobacco smoking: strong association of a specific major histocompatibility complex haplotype on chromosome 6 with smoking behavior. AB - The genetic basis for addiction to tobacco smoking--particularly that of the perception of olfactory stimuli that may be important in reinforcing smoking addiction--is largely unknown. A cluster of genes for olfactory receptors is in close proximity to the MHC region on chromosome 6. Polymorphisms of MHC class III genes (RCCX modules, TNFA promoter polymorphisms) were determined in 101 healthy subjects and 232 coronary artery disease (CAD) patients from Hungary with defined tobacco smoking habits. A highly significant association between ever smoking (past + current smokers) and a specific MHC haplotype was observed (odds ratios = 2.14-4.13; P-values = 0.012 to <0.001). This haplotype is characterized by the presence of C4A null alleles and a solitary short C4B gene linked to the TNF2 allele of the promoter for TNFA gene. This haplotype occurred more frequently in the ever smokers than in the never smokers [odds ratio: 4.97 (1.96-12.62); P = 0.001], and such associations were stronger in women (odds ratio = 13.6) than in men (odds ratio = 2.79). An independent study of complement C4 protein polymorphism and smoking habits in Icelandic subjects (n = 351) yielded similar and confirmative results. Considering the documented link between olfactory stimuli and smoking in females, and the presence of a cluster of odorant receptor genes close to the MHC class I region, our findings implicate a potential role of the MHC-linked olfactory receptor genes in the initiation of smoking. PMID- 15339883 TI - AILIM/ICOS signaling induces T-cell migration/polarization of memory/effector T cells. AB - AILIM/ICOS has critical roles in the regulation of T-cell differentiation and effector T-cell function in various immune responses. The counter-ligand for AILIM/ICOS, B7h, is widely expressed in not only lymphoid tissue and antigen presenting cells, but also in fibroblast and endothelial cells in various organs. Here, we demonstrate that activated human T-cells migrate beneath TNF-alpha treated HUVEC and display morphological polarization via AILIM/ICOS signaling. AILIM/ICOS stimulation, in the absence of antigen stimulation, also induced T cell polarization. Importantly, AILIM/ICOS-mediated polarization was evident in CD4+CD45RO+ memory T-cells and generated Th1 cells, but not in CD4+CD45RA+ naive T-cells and generated Th2 cells. Furthermore, AILIM/ICOS signaling is involved in transendothelial migration of Th1 cells, but not Th2 cells. Our data suggest that AILIM/ICOS-B7h interactions play an important role in the endothelium in controlling the entry of memory/effector T-cells into inflamed tissues in the periphery. PMID- 15339884 TI - A piece of my mind. Clinic. PMID- 15339885 TI - Antepartum dental radiography and low birth weight. PMID- 15339886 TI - Antepartum dental radiography and low birth weight. PMID- 15339887 TI - Antepartum dental radiography and low birth weight. PMID- 15339888 TI - Antepartum dental radiography and low birth weight. PMID- 15339889 TI - Antepartum dental radiography and low birth weight. PMID- 15339890 TI - Lead exposure and cognitive outcomes of children with prenatal cocaine exposure. PMID- 15339891 TI - Nicotine dependence and WHO mental health surveys. PMID- 15339892 TI - Educational programs in US medical schools, 2003-2004. AB - CONTEXT: US medical schools continue to change their organizational structures, staffing patterns, and educational programs. OBJECTIVE: To review the status of US medical school educational programs in the 2003-2004 academic year, compared with 1993-1994. DATA SOURCES: The Liaison Committee on Medical Education (LCME) Annual Medical School Questionnaire for 2003-2004 and the Association of American Medical Colleges Directory of American Medical Education for the years 1983-1984, 1993-1994, and 2003-2004. DATA SYNTHESIS: The number of full-time faculty members in the 126 LCME-accredited medical schools increased from 90 975 in 1993-1994 to 114 549 in 2003-2004 (+26%), whereas the number of enrolled students remained essentially unchanged (66 453 in 1993-1994 and 67 166 in 2003-2004). In 2003 2004, 48% of medical school deans held another title at the medical center or university level, such as vice president for health affairs. There are 94 medical schools that have a comprehensive clinical examination using the standardized patient/objective structured clinical examination format; 59 schools require students to pass this examination for graduation. As of spring 2004, 58 schools will require students in the class of 2005 to pass the new US Medical Licensing Examination Step 2 Clinical Skills examination. CONCLUSIONS: The role of the medical school dean has expanded over time and is associated with the creation of a discrete administrative structure for the educational program. The number of full-time medical school faculty continues to increase, whereas the number of enrolled students remains steady. Considerable variability exists among medical schools in their use of standardized clinical evaluations. PMID- 15339893 TI - US graduate medical education, 2003-2004. AB - CONTEXT: Information about recent graduates of medical schools and the characteristics of physicians training in graduate medical education (GME) portends the size and composition of the US physician workforce of the near future. OBJECTIVES: To examine trends in training programs and career choices of graduating male and female residents and to monitor trends in the size of the entire residency population. DESIGN, SETTING, AND PARTICIPANTS: The American Medical Association and Association of American Medical Colleges jointly surveyed residency programs during the academic year 2003-2004 about active, transferred, and graduated residents, as well as about program characteristics. The 8192 programs confirmed the status of 94.6% of residents. Nearly 86% of program directors (n = 7040) completed the accompanying program survey. MAIN OUTCOME MEASURES: Overall trends during the last 6 years in the number and characteristics of residents and programs, as well as the specialty of male and female graduating residents. RESULTS: There were 99,964 active residents during the 2003-2004 academic year, the highest ever recorded by the National GME Census. The number of residents (n = 22,444) entering US graduate medical education programs for the first time is also the highest on record. In 1999, 28,773 physicians completed training, 10,546 (36.7%) of whom were women. In 2003, there were 29,745 graduates, 11,681 (39.3%) of whom were women, representing a 10.8% increase. The number of obstetrics/gynecology male graduates decreased 31.3%, while female graduates increased 18.2%. Other specialties that lost men and gained women were dermatology, family medicine, internal medicine, ophthalmology, pathology, psychiatry, and general surgery. The proportion of graduates who pursued additional training increased; percentages were 27.2% in 1999, 29.6% in 2001, and 32.1% in 2003. In 2000, 35.7% of programs provided opportunities to develop cultural competence; the percentage in 2003 was 50.7%. The percentage of programs with complementary/alternative medicine curriculum has held steady at 24%. CONCLUSIONS: The number of physicians in GME is at its highest, and nearly one third of physicians completing training in one program continue on in another. The choices of female residents parallel those of male residents in many respects, but there are important differences. PMID- 15339894 TI - The role of physician specialty board certification status in the quality movement. AB - The Institute of Medicine's reports and discussions on quality of medical care have focused on a systems-based approach to quality improvement. Our objective is to summarize evidence and theory about the role of a physician's current board certification status in quality improvement. The first body of evidence includes the validity of board certification demonstrated by the testing process, the relationship of examination scores with other measures of physician competence, and the relationship between certification status and clinical outcomes. The second body of evidence involves the adaptation of error prevention theory to medical care. Patient safety is enhanced when problem-solving uses readily accessed habits of behavior, the same behavior necessary to achieve board certification. The third body of evidence, obtained through a Gallup poll, demonstrates that certification and maintenance of certification are highly valued by the public. The majority of respondents thought it important for physicians to be reevaluated on their qualifications every few years and that physicians should do more to demonstrate ongoing competence than is currently required by the profession. We conclude that a physician's current certification status should be among the evidence-based measures used in the quality movement. PMID- 15339895 TI - Educational epidemiology: applying population-based design and analytic approaches to study medical education. AB - Conducting educational research in medical schools is challenging partly because interventional controlled research designs are difficult to apply. In addition, strict accreditation requirements and student/faculty concerns about educational inequality reduce the flexibility needed to plan and execute educational experiments. Consequently, there is a paucity of rigorous and generalizable educational research to provide an evidence-guided foundation to support educational effectiveness. "Educational epidemiology," ie, the application across the physician education continuum of observational designs (eg, cross-sectional, longitudinal, cohort, and case-control studies) and randomized experimental designs (eg, randomized controlled trials, randomized crossover designs), could revolutionize the conduct of research in medical education. Furthermore, the creation of a comprehensive national network of educational epidemiologists could enhance collaboration and the development of a strong educational research foundation. PMID- 15339896 TI - Professional liability issues in graduate medical education. AB - Resident physicians, attending physicians, and graduate medical education (GME) institutions share a collective responsibility to deliver safe and appropriate care to patients. The law does not offer concessions in quality of care to accommodate GME. Resident physicians are generally held to the same standard of care as attending physicians in their respective specialties. This principle encourages resident physicians to seek supervision and attending physicians to provide the same. Attending physicians face malpractice exposure not only for the care they provide but also for the care they direct. In addition, they may be held vicariously liable for the negligence of resident physicians working with them, or directly liable for inadequate supervision. What constitutes adequate supervision is unsettled in the law. As the standards in this area evolve, it is reasonable to expect that the profession's and the public's heightened attention to patient safety will continue to move the courts toward higher standards of supervision. GME institutions and programs bear legal responsibility for both the care they deliver and the negligence of their employees. They also face liability for failing to administer safe systems of care. Work hour restrictions and a growing understanding of the role of organizational factors in contributing to and preventing medical injury may increase the legal expectations imposed on GME programs. PMID- 15339897 TI - Chronic disease--the need for a new clinical education. PMID- 15339898 TI - Professors not professing. PMID- 15339899 TI - Do tumors detected by mammography screening have a favorable prognosis? PMID- 15339900 TI - Risk for distant recurrence of breast cancer detected by mammography screening or other methods. AB - CONTEXT: Selection of systemic adjuvant therapies for women diagnosed as having breast cancer is based on risk estimations for cancer recurrence. In such estimations, tumors detected by mammography screening are considered to be associated with a similar risk of recurrence as tumors of similar size found by other methods. OBJECTIVE: To compare the risk of recurrence and survival among women with cancerous tumors detected by mammography screening compared with other methods (outside of screening). DESIGN, SETTING, AND PATIENTS: Retrospective study comparing clinical, histopathological, and biological features of cancerous tumors detected by mammography screening compared with tumors detected outside of screening. Women diagnosed as having breast cancer in 1991 or 1992 were identified from the Finnish Cancer Registry (n = 2842). The median follow-up time was 9.5 years. Cancer biological variables were analyzed from tumor tissue microarrays using immunohistochemistry or in situ hybridization and included ERBB2, TP53, and MK167 expression and ERBB2 amplification data. MAIN OUTCOME MEASURES: Univariate and multivariate analyses of potential risk factors for distant recurrence of breast cancer and 10-year survival. RESULTS: Of the 1983 women with unilateral invasive breast cancer, data on tumor diameter were available for 1918 women. Women with cancerous tumors detected by mammography screening had better estimated 10-year distant disease-free survival than women with tumors found outside of screening (tumor size of < or =10 mm [n = 386] 92% vs 85% [P =.04]; 11-20 mm [n = 808] 88% vs 76% [P<.001]; 21-30 mm [n = 409] 86% vs 63% [P =.008]; >30 mm [n = 315] 68% vs 50% [P =.12], respectively). In a Cox multivariate model that included cancer biological factors, the relative hazard ratio for distant recurrence among women with tumors detected outside of screening (HR, 1.90; 95% confidence interval, 1.15-3.11) was significantly higher than among women with tumors detected by mammography screening (P =.01). Breast cancer diagnosis by mammography screening was an independent prognostic variable reducing the relative HR for distant recurrence. This effect was equal to or greater than the effect of 1-cm decrease in tumor diameter (HR, 1.20; 95% confidence interval, 1.10-1.31). CONCLUSIONS: Cancerous tumors detected by mammography screening are associated with a better prognosis than tumors of similar size found outside of screening. The risk of distant metastases is overestimated for women diagnosed as having cancer by mammography screening unless the method of detection is taken into account in risk estimations. PMID- 15339901 TI - Systolic hypertension in older persons. AB - CONTEXT: The Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure emphasizes the importance of systolic hypertension (SH), defined as systolic blood pressure (SBP) of at least 140 mm Hg and diastolic blood pressure of less than 90 mm Hg, in older persons (> or =60 years). OBJECTIVE: To systematically review the literature on clinical management of SH in older persons. DATA SOURCES: We performed a MEDLINE search of English-language literature from 1966-2004 to identify reports about SH in older persons, with particular emphasis on data from randomized clinical trials. STUDY SELECTION AND DATA EXTRACTION: We selected 1064 studies by using the search terms hypertension combined with the terms systole (or systolic) and aged. DATA SYNTHESIS: There is strong evidence from clinical trials to support the treatment of SH in older persons with SBP of at least 160 mm Hg. Large-scale trials to assess the value of antihypertensive therapy for older patients with SBP of 140 to 159 mm Hg have not been performed, and recommendations to treat these patients are based on observational studies that show a graded relationship of cardiovascular risk with increasing SBP. The studies most strongly support the use of thiazide diuretics and long-acting calcium channel blockers as first-line therapy to treat SH. CONCLUSIONS: Treatment of SH in older patients with SBP of at least 160 mm Hg is supported by strong evidence. The evidence available to support treatment of patients to the level of 140 mm Hg or those with baseline SBP of 140 to 159 mm Hg is less strong; thus, these treatment decisions should be more sensitive to patient preferences and tolerance of therapy. PMID- 15339902 TI - JAMA patient page. Academic health centers. PMID- 15339903 TI - The subunit b dimer of the FOF1-ATP synthase: interaction with F1-ATPase as deduced by site-specific spin-labeling. AB - We have used site-specific spin-labeling of single cysteine mutations within a water-soluble mutant of subunit b of the ATP synthase and employed electron spin resonance (ESR) spectroscopy to obtain information about the binding interactions of the b dimer with F1-ATPase. Interaction of b2 with a delta-depleted F1 (F1 delta) was also studied. The cysteine mutations used for spin-labeling were distributed throughout the cytosolic domain of the b subunit. In addition, each position between residues 101 and 114 of b was individually mutated to cysteine. All mutants were modified with a cysteine-reactive spin label. The room temperature ESR spectra of spin-labeled b2 in the presence of F1 or F1-delta when compared with the spectra of free b2 indicate a tight binding interaction between b2 and F1. The data suggest that b2 packs tightly to F1 between residues 80 and the C terminus but that there are segments of b2 within that region where packing interactions are quite loose. Two-dimensional gel electrophoresis confirmed binding of the modified b mutants to F1-ATPase as well as to F1-delta. Subsequent addition of delta to F1-delta.b2 complex resulted in changes in the ESR spectra, indicating different binding interactions of b to F1 in the presence or absence of delta. The data also suggest that the reconstitution of the ATP synthase is not ordered with respect to these subunits. Additional spectral components observed in b preparations that were spin-labeled between amino acid position 101 and 114 are indicative of either two populations of b subunits with different packing interactions or to helical bending within this region. PMID- 15339904 TI - Syntaxin-1A inhibits cardiac KATP channels by its actions on nucleotide binding folds 1 and 2 of sulfonylurea receptor 2A. AB - ATP-sensitive potassium (KATP) channels couple the metabolic status of the cell to its membrane potential to regulate a number of cell actions, including secretion (neurons and neuroendocrine cells) and muscle contractility (skeletal, cardiac, and vascular smooth muscle). KATP channels consist of regulatory sulfonylurea receptors (SUR) and pore-forming (Kir6.X) subunits. We recently reported (Pasyk, E. A., Kang, Y., Huang, X., Cui, N., Sheu, L., and Gaisano, H. Y. (2004) J. Biol. Chem. 279, 4234-4240) that syntaxin-1A (Syn-1A), known to mediate exocytotic fusion, was capable of binding the nucleotide binding folds (NBF1 and C-terminal NBF2) of SUR1 to inhibit the KATP channels in insulin secreting pancreatic islet beta cells. This prompted us to examine whether Syn-1A might modulate cardiac SUR2A/KATP channels. Here, we show that Syn-1A is present in the plasma membrane of rat cardiac myocytes and binds the SUR2A protein (of rat brain, heart, and human embryonic kidney 293 cells expressing SUR2A/Kir6. 2) at its NBF1 and NBF2 domains to decrease KATP channel activation. Unlike islet beta cells, in which Syn-1A inhibition of the channel activity was apparently mediated only via NBF1 and not NBF2 of SUR1, both exogenous recombinant NBF1 and NBF2 of SUR2A were found to abolish the inhibitory actions of Syn-1A on K(ATP) channels in rat cardiac myocytes and HEK293 cells expressing SUR2A/Kir6.2. Together with our recent report, this study suggests that Syn-1A binds both NBFs of SUR1 and SUR2A but appears to exhibit distinct interactions with NBF2 of these SUR proteins in modulating the KATP channels in islet beta cells and cardiac myocytes. PMID- 15339905 TI - Activation state of the Ras2 protein and glucose-induced signaling in Saccharomyces cerevisiae. AB - The activity of adenylate cyclase in the yeast Saccharomyces cerevisiae is controlled by two G-protein systems, the Ras proteins and the Galpha protein Gpa2. Glucose activation of cAMP synthesis is thought to be mediated by Gpa2 and its G-protein-coupled receptor Gpr1. Using a sensitive GTP-loading assay for Ras2 we demonstrate that glucose addition also triggers a fast increase in the GTP loading state of Ras2 concomitant with the glucose-induced increase in cAMP. This increase is severely delayed in a strain lacking Cdc25, the guanine nucleotide exchange factor for Ras proteins. Deletion of the Ras-GAPs IRA2 (alone or with IRA1) or the presence of RAS2Val19 allele causes constitutively high Ras GTP loading that no longer increases upon glucose addition. The glucose-induced increase in Ras2 GTP-loading is not dependent on Gpr1 or Gpa2. Deletion of these proteins causes higher GTP loading indicating that the two G-protein systems might directly or indirectly interact. Because deletion of GPR1 or GPA2 reduces the glucose-induced cAMP increase the observed enhancement of Ras2 GTP loading is not sufficient for full stimulation of cAMP synthesis. Glucose phosphorylation by glucokinase or the hexokinases is required for glucose-induced Ras2 GTP loading. These results indicate that glucose phosphorylation might sustain activation of cAMP synthesis by enhancing Ras2 GTP loading likely through inhibition of the Ira proteins. Strains with reduced feedback inhibition on cAMP synthesis also display elevated basal and induced Ras2 GTP loading consistent with the Ras2 protein acting as a target of the feedback-inhibition mechanism. PMID- 15339906 TI - Structurally distinct elements mediate internal ribosome entry within the 5' noncoding region of a voltage-gated potassium channel mRNA. AB - The approximately 1.2-kb 5'-noncoding region (5'-NCR) of mRNA species encoding mouse Kv1.4, a member of the Shaker-related subfamily of voltage-gated potassium channels, was shown to mediate internal ribosome entry in cells derived from brain, heart, and skeletal muscle, tissues known to express Kv1.4 mRNA species. We also show that the upstream approximately 1.0 kb and the downstream approximately 0.2 kb of the Kv1.4 5'-NCR independently mediated internal ribosome entry; however, separately, these sequences were less efficient in mediating internal ribosome entry than when together in the complete (and contiguous) 5' NCR. Using enzymatic structure probing, the 3'-most approximately 0.2 kb was predicted to form three distinct stem-loop structures (stem-loops X, Y, and Z) and two defined single-stranded regions (loops Psi and Omega) in the presence and absence of the upstream approximately 1.0 kb. Although the systematic deletion of sequences within the 3'-most approximately 0.2 kb resulted in distinct changes in expression, enzymatic structure probing indicated that local RNA folding was not completely altered. Structure probing analysis strongly suggested an interaction between stem-loop X and a downstream polypyrimidine tract; however, opposing changes in activity were observed when sequences within these two regions were independently deleted. Moreover, deletions correlating with positive as well as negative changes in expression altered RNase cleavage within stem-loop X, indicating that this structure may be an integral element. Therefore, these findings indicate that Kv1.4 expression is mediated through a complex interplay between many distinct RNA regions. PMID- 15339908 TI - Role of CL-100, a dual specificity phosphatase, in thrombin-induced endothelial cell activation. AB - Using a cDNA microarray screening approach, we have identified seven novel thrombin-responsive genes in human umbilical vein endothelial cells that were verifiable by Northern blot analysis. Among them CL-100, a dual-specificity phosphatase also known as MAP kinase phosphatase-1 (MKP-1), showed greatest induction by thrombin. Steady-state levels of CL-100 mRNA induction by thrombin peaked at 1 h and declined rapidly (t1/2 approximately 45 min). Induction by thrombin was protease-activated receptor-1-mediated, protein synthesis independent, and transcriptionally regulated. Metabolic labeling followed by immunoprecipitation verified that the thrombin-induced CL-100 mRNA was translated into protein. We found that both Src-kinase and p42/p44 ERK activity are critical for thrombin-induced CL-100 expression, whereas phosphatidylinositol 3-kinase and protein kinase C activity were not required. Antisense-mediated inhibition of CL 100 was shown to prolong thrombin-induced ERK activity in endothelial cells, concomitant with an inhibition in thrombin-induced PDGF-A (platelet-derived growth factor A) and PDGF-B gene expression and an up-regulation in thrombin induced VCAM-1 and E-selectin gene expression. Inhibition of ERK activation by PD98059 in endothelial cells was shown to potentiate thrombin-induced expression of PDGF-B (approximately 3-fold) while inhibiting thrombin-induced VCAM-1 and E selectin gene expression by 60 and 70%, respectively. These results suggested that induced expression of the CL-100 phosphatase and its subsequent regulation of ERK activity play a key regulatory role in the thrombin signaling pathway and in the transcriptional regulation of pathologically important "endothelial cell activation genes." PMID- 15339909 TI - Ganglioside GD3 traffics from the trans-Golgi network to plasma membrane by a Rab11-independent and brefeldin A-insensitive exocytic pathway. AB - Gangliosides, complex glycosphingolipids containing sialic acids, have been found to reside in glycosphingolipid-enriched microdomains (GEM) at the plasma membrane. They are synthesized in the lumen of the Golgi complex and appear unable to translocate from the lumenal toward the cytosolic surface of Golgi membrane to access the monomeric lipid transport. As a consequence, they can only leave the Golgi complex via the lumenal surface of transport vesicles. In this work we analyzed the exocytic transport of the disialo ganglioside GD3 from trans Golgi network (TGN) to plasma membrane in CHO-K1 cells by immunodetection of endogenously synthesized GD3. We found that ganglioside GD3, unlike another luminal membrane-bounded lipid (glycosylphosphatidylinositol-anchored protein), did not partition into GEM domains in the Golgi complex and trafficked from TGN to plasma membrane by a brefeldin A-insensitive exocytic pathway. Moreover, a dominant negative form of Rab11, which prevents exit of vesicular stomatitis virus glycoprotein from the Golgi complex, did not influence the capacity of GD3 to reach the cell surface. Our results strongly support the notion that most ganglioside GD3 traffics from the TGN to the plasma membrane by a non conventional vesicular pathway where lateral membrane segregation of vesicular stomatitis virus glycoprotein (non-GEM resident) and glycosylphosphatidylinositol anchored proteins (GEM resident) from GD3 is required before exiting TGN. PMID- 15339910 TI - Sensing bacterial flagellin by membrane and soluble orthologs of Toll-like receptor 5 in rainbow trout (Onchorhynchus mikiss). AB - Rainbow trout (Onchorhynchus mikiss) possess two genes encoding putative leucine rich repeat (LRR)-containing proteins similar to human TLR5. Molecular cloning of these two LRR proteins suggested the presence of a TLR5-like membrane form (rtTLR5M) and a soluble form (rtTLR5S). Here we elucidated the primary structures and the unique combinational functions of these fish versions of TLR5. The LRR regions of rtTLR5S and rtTLR5M exhibited 81% homology and relatively high (35.6 and 33.7%) homology to the extracellular domains of human TLR5 (huTLR5). Thus, two distinct genes encode the TLR5 orthologs in fish, one of which has a consensus intracellular domain (TIR). In order to test their functions, we constructed fusion proteins with the LRR region of rtTLR5S (S-chimera) or that of rtTLR5M and the TIR of huTLR5 (M-chimera). The S- and M-chimeras expressed in HeLa or CHO cells signaled the presence of Vibrio anguillarum flagellin, resulting in NF-kappaB activation. rtTLR5M was ubiquitously expressed, whereas rtTLR5S was predominantly expressed in the liver. In the hepatoma cell lines of the rainbow trout RTH-149, stimulation of rtTLR5M with V. anguillarum or its flagellin allowed the up-regulation of rtTLR5S. Flagellin-mediated NF-kappaB activation was more significant in the presence of or simultaneous expression of rtTLR5S. Therefore, a two-step flagellin response occurred for host defense against bacterial infection in fish: (a) flagellin first induced basal activation of NF-kappaB via membrane TLR5, facilitating the production of soluble TLR5 and minimal acute phase proteins, and (b) the inducible soluble TLR5 amplifies membrane TLR5-mediated cellular responses in a positive feedback fashion. PMID- 15339911 TI - Critical role of endogenous Akt/IAPs and MEK1/ERK pathways in counteracting endoplasmic reticulum stress-induced cell death. AB - Endoplasmic reticulum (ER) stress has been implicated in the pathogenesis of many diseases and in cancer therapy. Although the unfolded protein response is known to alleviate ER stress by reducing the accumulation of misfolded proteins, the exact survival elements and their downstream signaling pathways that directly counteract ER stress-stimulated apoptotic signaling remain elusive. Here, we have shown that endogenous Akt and ERK are rapidly activated and act as downstream effectors of phosphatidylinositol 3-kinase in thapsigargin- or tunicamycin induced ER stress. Introduction of either dominant-negative Akt or MEK1 or the inhibitors LY294002 and U0126 sensitized cells to ER stress-induced cell death in different cell types. Reverse transcription-PCR analysis of gene expression during ER stress revealed that cIAP-2 and XIAP, members of the IAP family of potent caspase suppressors, were strongly induced. Transcription of cIAP-2 and XIAP was up-regulated by the phosphatidylinositol 3-kinase/Akt pathway as shown by its reversal by dominant-negative Akt or LY294002. Ablation of these IAPs by RNA interference sensitized cells to ER stress-induced death, which was reversed by the caspase inhibitor benzyloxycarbonyl-VAD-fluoromethyl ketone. The protective role of IAPs in ER stress coincided with Smac release from mitochondria to the cytosol. Furthermore, it was shown that mTOR was not required for Akt-mediated survival. These results represent the first demonstration that activation of endogenous Akt/IAPs and MEK/ERK plays a critical role in controlling cell survival by resisting ER stress-induced cell death signaling. PMID- 15339912 TI - Divergent effects of peroxisome proliferator-activated receptor gamma agonists and tumor necrosis factor alpha on adipocyte ApoE expression. AB - ApoE is expressed in multiple mammalian cell types in which it supports cellular differentiated function. In this report we demonstrate that apoE expression in adipocytes is regulated by factors involved in modulating systemic insulin sensitivity. Systemic treatment with pioglitazone increased systemic insulin sensitivity and increased apoE mRNA levels in adipose tissue by 2-3-fold. Treatment of cultured 3T3-L1 adipocytes with ciglitazone increased apoE mRNA levels by 2-4-fold in a dose-dependent manner and increased apoE secretion from cells. Conversely, treatment of adipocytes with tumor necrosis factor (TNF) alpha reduced apoE mRNA levels and apoE secretion by 60%. Neither insulin nor a peroxisome proliferator-activated receptor (PPAR) alpha agonist regulated adipocyte apoE gene expression. In addition, treatment of human monocyte-derived macrophages with ciglitazone did not regulate expression of apoE. Additional analyses using reporter genes indicated that the effect of TNFalpha and PPARgamma agonists on the apoE gene was mediated via distinct gene control elements. The TNFalpha effect was mediated by elements within the proximal promoter, whereas the PPARgamma effect was mediated by elements within a downstream enhancer. However, the addition of TNFalpha substantially reduced the absolute levels of apoE reporter gene response even in the presence of ciglitazone. These results indicate for the first time that adipose tissue expression of apoE is modulated by physiologic regulators of insulin sensitivity. PMID- 15339913 TI - A novel Galphaq/11-selective inhibitor. AB - YM-254890, which was isolated from the culture broth of Chromobacterium sp., inhibits ADP-induced platelet aggregation and has antithrombotic and thrombolytic effects. YM-254890 blocks Galpha(q/11)-coupled ADP receptor P2Y1-mediated Ca(2+) mobilization. Here we report that YM-254890 is a selective Galpha(q/11) inhibitor. YM-254890 blocked Ca(2+) mobilization mediated by several Galpha(q/11) coupled receptors but not by Galpha(i)- or Galpha(15)-coupled receptor, indicating that phospholipase Cbeta activation and subsequent signaling molecules are not the target of YM-254890. YM-254890 completely prevented the serum response factor (SRF)-mediated gene transcription induced by Galpha(q)R183C, which is constitutively active in a receptor-dependent manner because of its reduced k(cat) of GTP hydrolysis. Conversely, YM-254890 had only a modest effect on the SRF-mediated gene transcription by Galpha(q)Q209L, which is GTPase deficient (activated) Galpha(q). These suggested that the acting point of YM 254890 is receptor-Galpha(q) interaction or the subsequent guanine nucleotide exchange step. The fact that YM-254890 (i) inhibited the SRF-mediated gene transcription by Galpha(qi5), which interacts with Galpha(i)-coupled receptor and possesses the effector function of Galpha(q), and (ii) had no effect on the K(d) value of high affinity [(3)H]2MeSADP binding to P2Y1, which reflects the agonist receptor-Galpha ternary complex, suggested that receptor-Galpha(q/11) interaction is not the target of YM-254890. On the other hand, specific [(35)S]GTPgammaS binding to Galpha(q/11) stimulated by the M1 muscarinic acetylcholine receptor and P2Y1 were inhibited by YM-254890. These data indicate that YM-254890 blocks the exchange of GDP for GTP in Galpha(q/11) activation. This novel Galpha(q/11) selective inhibitor is a promising and powerful tool for studying Galpha(q/11) protein activation, Galpha(q/11) -coupled receptor signaling, and Galpha(q/11) mediated biological events. PMID- 15339914 TI - MsbA transporter-dependent lipid A 1-dephosphorylation on the periplasmic surface of the inner membrane: topography of francisella novicida LpxE expressed in Escherichia coli. AB - The lipid A anchor of Francisella tularensis lipopolysaccharide (LPS) lacks both phosphate groups present in Escherichia coli lipid A. Membranes of Francisella novicida (an environmental strain related to F. tularensis) contain enzymes that dephosphorylate lipid A and its precursors at the 1- and 4'-positions. We now report the cloning and characterization of a membrane-bound phosphatase of F. novicida that selectively dephosphorylates the 1-position. By transferring an F. novicida genomic DNA library into E. coli and selecting for low level polymyxin resistance, we isolated FnlpxE as the structural gene for the 1-phosphatase, an inner membrane enzyme of 239 amino acid residues. Expression of FnlpxE in a heptose-deficient mutant of E. coli caused massive accumulation of a previously uncharacterized LPS molecule, identified by mass spectrometry as 1-dephospho-Kdo2 lipid A. The predicted periplasmic orientation of the FnLpxE active site suggested that LPS export might be required for 1-dephosphorylation of lipid A. LPS and phospholipid export depend on the activity of MsbA, an essential inner membrane ABC transporter. Expression of FnlpxE in the msbA temperature-sensitive E. coli mutant WD2 resulted in 90% 1-dephosphorylation of lipid A at the permissive temperature (30 degrees C). However, the 1-phosphate group of newly synthesized lipid A was not cleaved at the nonpermissive temperature (44 degrees C). Our findings provide the first direct evidence that lipid A 1 dephosphorylation catalyzed by LpxE occurs on the periplasmic surface of the inner membrane. PMID- 15339915 TI - Recurrent fever promotes Plasmodium falciparum development in human erythrocytes. AB - The human malarial parasite Plasmodium falciparum (Pf) is exposed to wide temperature fluctuations during its life cycle, ranging from 25 degrees C in the mosquito vector and 37 degrees C in humans to 41 degrees C during febrile episodes in the patient. The repeated occurrence of fever at regular intervals is a characteristic of human malaria. We have examined the influence of repeated exposure to elevated temperatures encountered during fever on the intraerythrocytic development of the parasite. Using flow cytometry, we show that repeated exposure to temperatures mimicking febrile episodes promotes parasite development in human erythrocytes. Heat shock-mediated cytoprotection and growth promotion is dependent on the heat shock protein 90 (PfHsp90) multi-chaperone complex. Inhibition of PfHsp90 function using geldanamycin attenuates temperature dependent progression from the ring to the trophozoite stage. Geldanamycin inhibits parasite development by disrupting the PfHsp90 complex consisting of PfHsp70, PfPP5, and tubulin, among other proteins. While explaining the contribution of febrile episodes to the pathogenesis of malaria, our results implicate temperature as an important environmental cue used by the parasite to coordinate its development in humans. PMID- 15339916 TI - New short splice variants of the human cardiac Cavbeta2 subunit: redefining the major functional motifs implemented in modulation of the Cav1.2 channel. AB - Two new short splice variants of the Ca2+ channel beta2 subunit were cloned from human heart poly(A)(+) mRNA. The 410-amino acid beta2f subunit is encoded by exons 1A, 2A, 3, 4, 12, 13, and 14 of the human Cavbeta2 gene and lacks the protein kinase A phosphorylation site, the beta-interaction domain (De Waard, M., Pragnell, M., and Campbell, K. P. (1994) Neuron 13, 495-503), 40% of the beta-SH3 domain, and 73% of the guanylate kinase domain of the putative membrane associated guanylate kinases module (McGee, A. W., Nunziato, D. A., Maltez, J. M., Prehoda, K. E., Pitt, G. S., and Bredt, D. S. (2004) Neuron 42, 89-99), and helix alpha3 of the alpha1-subunit binding pocket (Van Petegem F., Clark, K. A., Chatelain, F. C., and Minor, D. L., Jr. (2004) Nature 429, 671-675). The beta2g transcript has two potential initiation codons. With the second ATG codon, it generates the 164-amino acid beta2Deltag subunit encoded essentially by the distal part of exon 14, and thus beta2Deltag completely lacks any of the above motifs. Immunoprecipitation analysis confirmed stable association of beta2f and beta2Deltag with the alpha1C subunit. The plasma membrane localization of beta2f and beta2Deltag was substantially increased by co-expression of the alpha1C,77 and alpha2delta subunits. In COS1 cells, beta2f and beta2Deltag increased plasma membrane targeting of the pore-forming alpha1C subunit and differentially facilitated (beta2f > beta2Deltag) the voltage gating of otherwise silent Cav1.2 channels. We conclude that it is unlikely that the beta-interaction domain, membrane-associated guanylate kinases module, and the alpha1-subunit binding pocket helix alpha3 are essential for the interaction of the alpha1C and beta2 subunits and suggest that in addition to the alpha1-subunit binding pocket helices alpha5 and alpha8, a yet unresolved C-terminal beta2 region plays a crucial role. PMID- 15339917 TI - Leukotriene A4 hydrolase/aminopeptidase, the gatekeeper of chemotactic leukotriene B4 biosynthesis. PMID- 15339918 TI - Identification of crucial histidines involved in carbon-nitrogen triple bond synthesis by aldoxime dehydratase. AB - Aldoxime dehydratase (OxdA), which is a novel heme protein, catalyzes the dehydration of an aldoxime to a nitrile even in the presence of water in the reaction mixture. The combination of site-directed mutagenesis of OxdA (mutation of all conserved histidines in the aldoxime dehydratase superfamily), estimation of the heme contents and specific activities of the mutants, and CD and resonance Raman spectroscopic analyses led to the identification of the proximal and distal histidines in this unique enzyme. The heme contents and CD spectra in the far-UV region of all mutants except for the H299A one were almost identical to those of the wild-type OxdA, whereas the H299A mutant lost the ability of binding heme, demonstrating that His(299) is the proximal histidine. On the other hand, substitution of alanine for His(320) did not affect the overall structure of OxdA but caused loss of its ability of carbon-nitrogen triple bond synthesis and a lower shift of the Fe-C stretching band in the resonance Raman spectrum for the CO-bound form. Furthermore, the pH dependence of the wild-type OxdA closely followed the His protonation curves observed for other proteins. These findings suggest that His(320) is located in the distal heme pocket of OxdA and would donate a proton to the substrate in the aldoxime dehydration mechanism. PMID- 15339919 TI - Small heat shock protein alphaB-crystallin is part of cell cycle-dependent Golgi reorganization. AB - AlphaB-crystallin is a developmentally regulated small heat shock protein known for its binding to a variety of denatured polypeptides and suppression of protein aggregation in vitro. Elevated levels of alphaB-crystallin are known to be associated with a number of neurodegenerative pathologies such as Alzheimer disease and multiple sclerosis. Mutations in alphaB-crystallin gene have been linked to desmin related cardiomyopathy and cataractogenesis. The physiological function of this protein, however, is unknown. Using discontinuous sucrose density gradient fractionation of post-nuclear supernatants, prepared from rat tissues and human glioblastoma cell line U373MG, we have identified discrete membrane-bound fractions of alphaB-crystallin, which co-sediment with the Golgi matrix protein, GM130. Confocal microscopy reveals co-localization of alphaB crystallin with BODIPY TR ceramide and the Golgi matrix protein, GM130, in the perinuclear Golgi in human glioblastoma U373MG cells. Examination of synchronized cultures indicated that alphaB-crystallin follows disassembly of the Golgi at prometaphase and its reassembly at the completion of cytokinesis, suggesting that this small heat shock protein, with its chaperone-like activity, may have an important role in the Golgi reorganization during cell division. PMID- 15339920 TI - Effects of cardiotrophin on adipocytes. AB - Cardiotrophin (CT-1) is a naturally occurring protein member of the interleukin (IL)-6 cytokine family and signals through the gp130/leukemia inhibitory factor receptor (LIFR) heterodimer. The formation of gp130/LIFR complex triggers the auto/trans-phosphorylation of associated Janus kinases, leading to the activation of Janus kinase/STAT and MAPK (ERK1 and -2) signaling pathways. Since adipocytes express both gp130 and LIFR proteins and are responsive to other IL-6 family cytokines, we examined the effects of CT-1 on 3T3-L1 adipocytes. Our studies have shown that CT-1 administration results in a dose- and time-dependent activation and nuclear translocation of STAT1, -3, -5A, and -5B as well as ERK1 and -2. We also confirmed the ability of CT-1 to induce signaling in fat cells in vivo. Our studies revealed that neither CT-1 nor ciliary neurotrophic factor treatment affected adipocyte differentiation. However, acute CT-1 treatment caused an increase in SOCS-3 mRNA in adipocytes and a transient decrease in peroxisome proliferator-activated receptor gamma (PPARgamma) mRNA that was regulated by the binding of STAT1 to the PPARgamma2 promoter. The effects of CT-1 on SOCS-3 and PPARgamma mRNA were independent of MAPK activation. Chronic administration of CT 1 to 3T3-L1 adipocytes resulted in a decrease of both fatty acid synthase and insulin receptor substrate-1 protein expression yet did not effect the expression of a variety of other adipocyte proteins. Moreover, chronic CT-1 treatment resulted in the development of insulin resistance as judged by a decrease in insulin-stimulated glucose uptake. In summary, CT-1 is a potent regulator of signaling in adipocytes in vitro and in vivo, and our current efforts are focused on determining the role of this cardioprotective cytokine on adipocyte physiology. PMID- 15339921 TI - The NS5A protein of hepatitis C virus is a zinc metalloprotein. AB - The NS5A protein of hepatitis C virus is believed to be an integral part of the viral replicase. Despite extensive investigation, the role of this protein remains elusive. Only limited biochemical characterization of NS5A has been performed, with most research to date involving the myriad of host proteins and signaling cascades that interact with NS5A. The need for better characterization of NS5A is paramount for elucidating the role of this protein in the virus life cycle. Examination of NS5A using bioinformatics tools suggested the protein consisted of three domains and contained an unconventional zinc binding motif within the N-terminal domain. We have developed a method to produce NS5A and performed limited proteolysis to confirm the domain organization model. The zinc content of purified NS5A and the N-terminal domain of NS5A was determined, and each of these proteins was found to coordinate one zinc atom per protein. The predicted zinc binding motif consists of four cysteine residues, conserved among the Hepacivirus and Pestivirus genera, fitting the formula of CX17CXCX20C. Mutation of any of the four cysteine components of this motif reduced NS5A zinc coordination and led to a lethal phenotype for HCV RNA replication, whereas mutation of other potential metal coordination residues in the N-terminal domain of NS5A, but outside the zinc binding motif, had little effect on zinc binding and, aside from one exception, were tolerated for replication. Collectively, these results indicate that NS5A is a zinc metalloprotein and that zinc coordination is likely required for NS5A function in the hepatitis C replicase. PMID- 15339922 TI - Mutational analysis of arginine 276 in the leucine-loop of human uracil-DNA glycosylase. AB - Uracil residues are eliminated from cellular DNA by uracil-DNA glycosylase, which cleaves the N-glycosylic bond between the uracil base and deoxyribose to initiate the uracil-DNA base excision repair pathway. Co-crystal structures of the core catalytic domain of human uracil-DNA glycosylase in complex with uracil containing DNA suggested that arginine 276 in the highly conserved leucine intercalation loop may be important to enzyme interactions with DNA. To investigate further the role of Arg(276) in enzyme-DNA interactions, PCR-based codon-specific random mutagenesis, and site-specific mutagenesis were performed to construct a library of 18 amino acid changes at Arg(276). All of the R276X mutant proteins formed a stable complex with the uracil-DNA glycosylase inhibitor protein in vitro, indicating that the active site structure of the mutant enzymes was not perturbed. The catalytic activity of the R276X preparations was reduced; the least active mutant, R276E, exhibited 0.6% of wildtype activity, whereas the most active mutant, R276H, exhibited 43%. Equilibrium binding studies utilizing a 2-aminopurine deoxypseudouridine DNA substrate showed that all R276X mutants displayed greatly reduced base flipping/DNA binding. However, the efficiency of UV-catalyzed cross-linking of the R276X mutants to single-stranded DNA was much less compromised. Using a concatemeric [(32)P]U.A DNA polynucleotide substrate to assess enzyme processivity, human uracil-DNA glycosylase was shown to use a processive search mechanism to locate successive uracil residues, and Arg(276) mutations did not alter this attribute. PMID- 15339923 TI - To slip or skip, visualizing frameshift mutation dynamics for error-prone DNA polymerases. AB - Three models describing frameshift mutations are "classical" Streisinger slippage, proposed for repetitive DNA, and "misincorporatation misalignment" and "dNTP-stabilized misalignment," proposed for non-repetitive DNA. We distinguish between models using pre-steady state fluorescence kinetics to visualize transiently misaligned DNA intermediates and nucleotide incorporation products formed by DNA polymerases adept at making small frameshift mutations in vivo. Human polymerase (pol) mu catalyzes Streisinger slippage exclusively in repetitive DNA, requiring as little as a dinucleotide repeat. Escherichia coli pol IV uses dNTP-stabilized misalignment in identical repetitive DNA sequences, revealing that pol mu and pol IV use different mechanisms in repetitive DNA to achieve the same mutational end point. In non-repeat sequences, pol mu switches to dNTP-stabilized misalignment. pol beta generates -1 frameshifts in "long" repeats and base substitutions in "short" repeats. Thus, two polymerases can use two different frameshift mechanisms on identical sequences, whereas one polymerase can alternate between frameshift mechanisms to process different sequences. PMID- 15339924 TI - Galpha13 stimulates cell migration through cortactin-interacting protein Hax-1. AB - Galpha13, the alpha-subunit of the heterotrimeric G protein G13, has been shown to stimulate cell migration in addition to inducing oncogenic transformation. Cta, a Drosophila ortholog of G13, has been shown to be critical for cell migration leading to the ventral furrow formation in Drosophila embryos. Loss of Galpha13 has been shown to disrupt cell migration associated with angiogenesis in developing mouse embryos. Whereas these observations point to the vital role of G13-orthologs in regulating cell migration, widely across the species barrier, the mechanism by which Galpha13 couples to cytoskeleton and cell migration is largely unknown. Here we show that Galpha13 physically interacts with Hax-1, a cytoskeleton-associated, cortactin-interacting intracellular protein, and this interaction is required for Galpha13-stimulated cell migration. Hax-1 interaction is specific to Galpha13, and this interaction is more pronounced with the mutationally or functionally activated form of Galpha13 as compared with the wild type Galpha13. Expression of Hax-1 reduces the formation of actin stress fibers and focal adhesion complexes in Galpha13-expressing NIH3T3 cells. Coexpression of Hax-1 also attenuates Galpha(13)-stimulated activity of Rho while potentiating Galpha13-stimulated activity of Rac. The presence of a quadnary complex consisting of Galpha13, Hax-1, Rac, and cortactin indicates the role of Hax-1 in tethering Galpha13 to the cytoskeletal component(s) involved in cell movement. Whereas the expression of Hax-1 potentiates Galpha13-mediated cell movement, silencing of endogenous Hax-1 with Hax-1-specific small interfering RNAs drastically reduces Galpha13-mediated cell migration. These findings, along with the observation that Hax-1 is overexpressed in metastatic tumors and tumor cell lines, suggest a novel role for the association of oncogenic Galpha13 and Hax-1 in tumor metastasis. PMID- 15339925 TI - Augmentation of therapeutic angiogenesis using genetically modified human endothelial progenitor cells with altered glycogen synthase kinase-3beta activity. AB - Previously we reported that inhibition of glycogen synthase kinase-3beta (GSK3beta), a key regulator in many intracellular signaling pathways, enhances the survival and migration of vascular endothelial cells. Here we investigated the effect of inhibition of GSK3beta activity on the angiogenic function of endothelial progenitor cell (EPC) and demonstrated a new therapeutic angiogenesis strategy using genetically modified EPC. As we previously reported, two biologically distinct types of EPC, spindle-shaped "early EPC" and cobblestone shaped "late EPC" could be cultivated from human peripheral blood. Catalytically inactive GSK3beta gene was transduced into both EPC. Inhibition of GSK3beta signaling pathway led to increased nuclear translocation of beta-catenin and increased secretion of angiogenic cytokines (vascular endothelial growth factor and interleukin-8). It enhanced the survival and proliferation of early EPC, whereas it promoted the survival and differentiation of late EPC. Transplantation of either of these genetically modified EPC into the ischemic hind limb model of athymic nude mouse significantly improved blood flow, limb salvage, and tissue capillary density compared with nontransduced EPC. Inhibition of GSK3beta signaling of either of these genetically modified EPC augmented the in vitro and in vivo angiogenic potency of these cell populations. These data provide evidence that GSK3beta has a key role in the angiogenic properties of EPC. Furthermore, the genetic modification of EPC to alter this signaling step can improve the efficacy of cell-based therapeutic vasculogenesis. PMID- 15339926 TI - Molecular dissection of the FcRbeta signaling amplifier. AB - Human high affinity IgE receptors are expressed as two different isoforms: the tetrameric isoform, alphabetagamma(2), or the trimeric isoform, alphagamma(2). The alpha chain is the IgE binding subunit, whereas the FcRbeta and FcRgamma chains are the signaling modules. Both FcRbeta and FcRgamma contain immunoreceptor tyrosine-based activation motifs (ITAM), but the beta ITAM differs from canonical ITAMs in two ways; the spacing between the two canonical tyrosines harbors a third tyrosine, and it is one amino acid shorter than in canonical ITAMs, making it unfit to bind the tandem SH2 of Syk. We have shown that FcRbeta functions as an amplifier of the FcRgamma signaling function. However, the molecular mechanism of this amplification remains unclear. Here we show that mutation of the three tyrosines (Tyr-219, Tyr-225, and Tyr-229) in the beta ITAM essentially converts alphabetagamma(2)into an alphagamma(2) complex in terms of Lyn recruitment, FcRgamma phosphorylation, Syk activation, and calcium mobilization. Tyr-219 is the most critical residue in this regard. In addition, a detailed analysis of the dynamics of calcium mobilization suggests a possible inhibitory role for Tyr-225, which becomes apparent when Tyr-219 is mutated. Thus, the signaling amplification function of FcRbeta is mainly encoded in Tyr 219 and in its capacity to recruit Lyn. In turn, this Tyr-219-mediated Lyn recruitment enhances gamma chain phosphorylation, Syk activation, and calcium mobilization. The two other tyrosines appear to have a modulating function that remains to be fully assessed. PMID- 15339927 TI - Subcellular localization of multiple PREP2 isoforms is regulated by actin, tubulin, and nuclear export. AB - The PREP, MEIS, and PBX families are mammalian members of the TALE (three amino acid loop extension) class of homeodomain-containing transcription factors. These factors have been implicated in cooperative DNA binding with the HOX class of homeoproteins, but PREP and MEIS interact with PBX in apparently non-HOX dependent cooperative DNA binding as well. PREP, MEIS, and PBX have all been reported to reside in the cytoplasm in one or more tissues of the developing vertebrate embryo. In the case of PBX, cytoplasmic localization is due to the modulation of nuclear localization signals, nuclear export sequences, and interaction with a cytoplasmic anchoring factor, non-muscle myosin heavy chain II B. Here we report that murine PREP2 exists in multiple isoforms distinguished by interaction with affinity-purified antibodies raised to N- and C-terminal epitopes and by nuclear versus cytoplasmic localization. Alternative splicing gives rise to some of these PREP2 isoforms, including a 25-kDa variant lacking the C-terminal half of the protein and homeodomain and having the potential to act as dominant-negative. We further show that cytoplasmic localization is due to the concerted action of nuclear export, as evidenced by sensitivity to leptomycin B, and cytoplasmic retention by the actin and microtubule cytoskeletons. Cytoplasmic PREP2 colocalizes with both the actin and microtubule cytoskeletons and coimmunoprecipitates with actin and tubulin. Importantly, disruption of either cytoskeletal system redirects cytoplasmic PREP2 to the nucleus. We suggest that transcriptional regulation by PREP2 is modulated through the subcellular distribution of multiple isoforms and by interaction with two distinct cytoskeletal systems. PMID- 15339928 TI - The Dnmt1 DNA-(cytosine-C5)-methyltransferase methylates DNA processively with high preference for hemimethylated target sites. AB - In the cell, Dnmt1 is the major enzyme in maintenance of the pattern of DNA methylation after DNA replication. Evidence suggests that the protein is located at the replication fork, where it could directly modify nascent DNA immediately after replication. To elucidate the potential mechanism of this process, we investigate the processivity of DNA methylation and accuracy of copying an existing pattern of methylation in this study using purified Dnmt1 and hemimethylated substrate DNA. We demonstrate that Dnmt1 methylates a hemimethylated 958-mer substrate in a highly processive reaction. Fully methylated and unmethylated CG sites do not inhibit processive methylation of the DNA. Extending previous work, we show that unmethylated sites embedded in a hemimethylated context are modified at an approximately 24-fold reduced rate, which demonstrates that the enzyme accurately copies existing patterns of methylation. Completely unmodified DNA is methylated even more slowly due to an allosteric activation of Dnmt1 by methylcytosine-containing DNA. Interestingly, Dnmt1 is not able to methylate hemimethylated CG sites on different strands of the DNA in a processive manner, indicating that Dnmt1 keeps its orientation with respect to the DNA while methylating the CG sites on one strand of the DNA. PMID- 15339929 TI - The role of extra fragment at the C-terminal of cytochrome b (Residues 421-445) in the cytochrome bc1 complex from Rhodobacter sphaeroides. AB - Sequence alignment of cytochrome b of the cytochrome bc1 complex from various sources reveals that bacterial cytochrome b contain an extra fragment at the C terminus. To study the role of this fragment in bacterial cytochrome bc1 complex, Rhodobacter sphaeroides mutants expressing His-tagged cytochrome bc1 complexes with progressive deletion from this fragment (residues 421-445) were generated and characterized. The cytbDelta-(433-445) bc1 complex, in which 13 residues from the C-terminal end of this fragment are deleted, has electron transfer activity, subunit composition, and physical properties similar to those of the complement complex, indicating that this region of the extra fragment is not essential. In contrast, the electron transfer activity, binding of cytochrome b, ISP, and subunit IV to cytochrome c1, redox potentials of cytochromes b and c1 in the cytbDelta-(427-445), cytbDelta-(425-445), and cytbDelta-(421-445) mutant complexes, in which 19, 21, or all residues of this fragment are deleted, decrease progressively. EPR spectra of the [2Fe-2S] cluster and the cytochromes b in these three deletion mutant bc1 complexes are also altered; the extent of spectral alteration increases as this extra fragment is shortened. These results indicate that the first 12 residues (residues 421-432) from the N-terminal end of the C-terminal extra fragment of cytochrome b are essential for maintaining structural integrity of the bc1 complex. PMID- 15339930 TI - MiaB protein is a bifunctional radical-S-adenosylmethionine enzyme involved in thiolation and methylation of tRNA. AB - The last biosynthetic step for 2-methylthio-N(6)-isopentenyl-adenosine (ms(2)i(6)A), present at position 37 in some tRNAs, consists of the methylthiolation of the isopentenyl-adenosine (i(6)A) precursor. In this work we have reconstituted in vitro the conversion of i(6)A to ms(2)i(6)A within a tRNA substrate using the iron-sulfur MiaB protein, S-adenosylmethionine (AdoMet), and a reducing agent. We show that a synthetic i(6)A-containing RNA corresponding to the anticodon stem loop of tRNA(Phe) is also a substrate. This study demonstrates that MiaB protein is a bifunctional system, involved in both thiolation and methylation of i(6)A. In this process, one molecule of AdoMet is converted to 5' deoxyadenosine, probably through reductive cleavage and intermediate formation ofa5'-deoxyadenosyl radical as observed in other "Radical-AdoMet" enzymes, and a second molecule of AdoMet is used as a methyl donor as shown by labeling experiments. The origin of the sulfur atom is discussed. PMID- 15339931 TI - Protein kinase Cdelta activation induces apoptosis in response to cardiac ischemia and reperfusion damage: a mechanism involving BAD and the mitochondria. AB - Heart attacks caused by occlusion of coronary arteries are often treated by mechanical or enzymatic removal of the occlusion and reperfusion of the ischemic heart. It is now recognized that reperfusion per se contributes to myocardial damage, and there is a great interest in identifying the molecular basis of this damage. We recently showed that inhibiting protein kinase Cdelta (PKCdelta) protects the heart from ischemia and reperfusion-induced damage. Here, we demonstrate that PKCdelta activity and mitochondrial translocation at the onset of reperfusion mediates apoptosis by facilitating the accumulation and dephosphorylation of the pro-apoptotic BAD (Bcl-2-associated death promoter), dephosphorylation of Akt, cytochrome c release, PARP (poly(ADP-ribose) polymerase) cleavage, and DNA laddering. Our data suggest that PKCdelta activation has a critical proapoptotic role in cardiac responses following ischemia and reperfusion. PMID- 15339932 TI - Identification of a zinc finger domain in the human NEIL2 (Nei-like-2) protein. AB - The recently identified human NEIL2 (Nei-like-2) protein, a DNA glycosylase/AP lyase specific for oxidatively damaged bases, shares structural features and reaction mechanism with the Escherichia coli DNA glycosylases, Nei and Fpg. Amino acid sequence analysis of NEIL2 suggested it to have a zinc finger-like Nei/Fpg. However, the Cys-X2-His-X16-Cys-X2-Cys (CHCC) motif present near the C terminus of NEIL2 is distinct from the zinc finger motifs of Nei/Fpg, which are of the C4 type. Here we show the presence of an equimolar amount of zinc in NEIL2 by inductively coupled plasma mass spectrometry. Individual mutations of Cys-291, His-295, Cys-315, and Cys-318, candidate residues for coordinating zinc, inactivated the enzyme by abolishing its DNA binding activity. H295A and C318S mutants were also shown to lack bound zinc, and a significant change in their secondary structure was revealed by CD spectra analysis. Molecular modeling revealed Arg-310 of NEIL2 to be a critical residue in its zinc binding pocket, which is highly conserved throughout the Fpg/Nei family. A R310Q mutation significantly reduced the activity of NEIL2. We thereby conclude that the zinc finger motif in NEIL2 is essential for its structural integrity and enzyme activity. PMID- 15339933 TI - The transcriptional repressor hDaxx potentiates p53-dependent apoptosis. AB - p53 and its homologues p73 and p63 are transcription factors that play an essential role in modulating cell cycle arrest and cell death in response to several environmental stresses. The type and intensity of these responses, which can be different depending on the inducing stimulus and on the overall cellular context, are believed to rely on the activation of defined subsets of target genes. The proper activation of p53 family members requires the coordinated action of post-translational modifications and interaction with several cofactors. In this study, we demonstrate that the multifunctional protein hDaxx interacts with p53 and its homologues, both in vitro and in vivo, and modulates their transcriptional activity. Moreover, we show that hDaxx, which has been implicated in several apoptotic pathways, increases the sensitivity to DNA damage induced cell death and that this effect requires the presence of p53. Although hDaxx represses p53-dependent transcription of the p21 gene, it does not affect the activation of proapoptotic genes, and therefore acts by influencing the balance between cell cycle arrest and proapoptotic p53 targets. Our results therefore underline the central role of hDaxx in modulating the apoptotic threshold upon several stimuli and identify it as a possible integrating factor that coordinates the response of p53 family members. PMID- 15339934 TI - B-Raf contributes to sustained extracellular signal-regulated kinase activation associated with interleukin-2 production stimulated through the T cell receptor. AB - A T cell receptor (TCR) recognizes and responds to an antigenic peptide in the context of major histocompatibility complex-encoded molecules. This provokes T cells to produce interleukin-2 (IL-2) through extracellular signal-regulated kinase (ERK) activation. We investigated the roles of B-Raf in TCR-mediated IL-2 production coupled with ERK activation in the Jurkat human T cell line. We found that TCR cross-linking could induce up-regulation of both B-Raf and Raf-1 activities, but Raf-1 activity was decreased rapidly. On the other hand, TCR stimulated kinase activity of B-Raf was sustained. Expression of a dominant negative mutant of B-Raf abrogated sustained but not transient TCR-mediated MEK/ERK activation. The inhibition of sustained ERK activation by either expression of a dominant-negative B-Raf or treatment with a MEK inhibitor resulted in a decrease of the TCR-stimulated nuclear factor of activated T cells (NFAT) activity and IL-2 production. Collectively, our data provide the first direct evidence that B-Raf is a positive regulator of TCR-mediated sustained ERK activation, which is required for NFAT activation and the full production of IL 2. PMID- 15339935 TI - Inhibition of HIV-1 envelope glycoprotein-mediated cell fusion by a DL-amino acid containing fusion peptide: possible recognition of the fusion complex. AB - The N-terminal fusion peptide (FP) of human immunodeficiency virus-1 (HIV-1) is a potent inhibitor of cell-cell fusion, possibly because of its ability to recognize the corresponding segments inside the fusion complex within the membrane. Here we show that a fusion peptide in which the highly conserved Ile(4), Phe(8), Phe(11), and Ala(14) were replaced by their d-enantiomers (IFFA) is a potent inhibitor of cell-cell fusion. Fourier transform infrared spectroscopy confirmed that despite these drastic modifications, the peptide preserved most of its structure within the membrane. Fluorescence energy transfer studies demonstrated that the diastereomeric peptide interacted with the wild type FP, suggesting this segment as the target site for inhibition of membrane fusion. This is further supported by the similar localization of the wild type and IFFA FPs to microdomains in T cells and the preferred partitioning into ordered regions within sphingomyelin/phosphatidyl-choline/cholesterol giant vesicles. These studies provide insight into the mechanism of molecular recognition within the membrane milieu and may serve in designing novel HIV entry inhibitors. PMID- 15339936 TI - Homologous adaptation to oxidative stress induced by the photosensitized Pd bacteriochlorophyll derivative (WST11) in cultured endothelial cells. AB - Various forms of cellular stress induce adaptive responses through poorly understood mechanisms. In maintaining homeostasis, endothelial cells respond and adapt to changes in oxidative stress that prevail in the circulation. Endothelial cells are also the target of many oxidative stress-based vascular therapies. The objectives of this study were to determine whether endothelial cells adapt to oxidative stress induced upon the photosensitization of WST11 (a water-soluble Pd bacteriochlorophyll derivative being developed as a photodynamic agent) and to study possible cellular mechanisms involved. The hallmark of WST11-based photodynamic therapy is the in situ generation of cytotoxic reactive oxygen species causing vascular shutdown, hypoxia, and tumor eradication. Here we demonstrated that photodynamic therapy also induces adaptive responses and tolerance following a sublethal preconditioning of endothelial cells with the same (homologous) or different (heterologous) stressor. A link among p38 MAPK activity, expression of hsp70 and hsp27, and homologous adaptation to reactive oxygen species induced by photosensitized WST11 was established. In addition to characterization of some key proteins involved, our observations provide a beneficial new working tool for the studies of mechanisms involved in oxidative stress and adaptation using light-controlled photosensitization. PMID- 15339937 TI - Membrane retrieval in neutrophils during phagocytosis: inhibition by M protein expressing S. pyogenes bacteria. AB - During phagocytosis and phagosome maturation, complex membrane traffic events must be coordinated. We have observed, using fluorescent fluid-phase and membrane markers, that in the human neutrophil, internalization of nonopsonized, Gram positive bacteria, but not of latex beads, is accompanied by a rapid and localized formation of pinosomal structures. This pinocytic response is calcium dependent but insensitive to actin cytoskeleton disruption and wortmannin treatment. Contrary to what we observe, endosomal structures usually are considered to participate in phagosome formation by providing necessary membrane to forming phagosomes. Instead, our results show a coupling between neutrophil secretory and membrane-retrieval processes during phagosome maturation, and we suggest that the observed, localized pinocytic response is linked to the secretion of azurophilic granules toward nascent phagosomes. Accordingly, M and M like protein-expressing Streptococcus pyogenes bacteria, which are able to survive inside neutrophil phagosomes, inhibit both the secretion of azurophilic granules to phagosomes and pinosome formation. PMID- 15339938 TI - Pivotal roles of interleukin-6 in transmural inflammation in murine T cell transfer colitis. AB - Breakdown of normal mucosal immunity is one of the major causes for inflammatory bowel disease. Interleukin (IL)-6 is a proinflammatory cytokine produced aberrantly in various types of inflammation, but its role in inflammatory bowel disease is still obscure. Hence, we analyzed the roles of IL-6 in the pathogenesis of murine T cell transfer colitis, whose histopathology resembles Crohn's disease. The transfer of CD4+CD45RBhigh T cells into severe combined immunodeficiency mice induced the infiltration of T cells and macrophages, and the gene expression of CC chemokine receptor (CCR)1, CCR2, CCR5, CXC chemokine receptor 3, their ligands, tumor necrosis factor-alpha, interferon-gamma, and IL 6 was progressively augmented as colitis developed. The incidence of transmural colitis was significantly reduced with a minimal decrease in the severity of colitis in recipients transferred with CD4+CD45RBhigh T cells derived from IL-6 deficient mice compared with those with wild-type mice. Moreover, the gene expression of several cytokines, chemokines, and matrix metalloproteinases was reduced significantly in recipients transferred with IL-6-deficient, mice-derived T cells. These observations suggested that T cell-derived IL-6 may augment the gene expression of several proinflammatory molecules, thereby causing transmural inflammation. Thus, IL-6 might be a promising target for treating transmural inflammation in Crohn's disease, which can lead to severe complications such as strictures, fissures, and fistulas. PMID- 15339940 TI - Myoglobin: an essential hemoprotein in striated muscle. AB - Myoglobin is a cytoplasmic hemoprotein, expressed solely in cardiac myocytes and oxidative skeletal muscle fibers, that reversibly binds O2 by its heme residue, a porphyrin ring:iron ion complex. Since the initial discovery of its structure over 40 years ago, wide-ranging work by many investigators has added importantly to our understanding of its function and regulation. Functionally, myoglobin is well accepted as an O2-storage protein in muscle, capable of releasing O2 during periods of hypoxia or anoxia. Myoglobin is also thought to buffer intracellular O2 concentration when muscle activity increases and to facilitate intracellular O2 diffusion by providing a parallel path that augments simple diffusion of dissolved O2. The use of gene targeting and other molecular biological techniques has revealed important new insights into the developmental and environmental regulation of myoglobin and provided additional functions for this hemoprotein such as scavenging nitric oxide and reactive O2 species. These recent findings, coupled with additional emerging technologies and the discovery of other tissue globins, provide a framework for addressing new questions about myoglobin and readdressing old ones. PMID- 15339939 TI - Natural killer cells inhibit hepatitis C virus expression. AB - Natural killer (NK) cells are critical in host innate defense against certain viruses. The role of NK cells in controlling hepatitis C virus (HCV) remains obscure. We examined whether NK cells are capable of inhibiting HCV expression in human hepatic cells. When NK cells are cultured with the HCV replicon-containing hepatic cells, they have no direct cytolytic effect but release soluble factor(s) suppressing HCV RNA expression. Media conditioned by NK cell lines (NK-92 and YTS) or primary NK cells isolated from healthy donors contain interferon gamma (IFN-gamma) and potently inhibit HCV RNA expression. Ligation of CD81 on NK cells inhibits IFN-gamma production and results in decreased anti-HCV activity. In addition, the antibodies to IFN-gamma or IFN-gamma receptors abolish the anti-HCV activity of NK cell-conditioned media. The role of IFN-gamma in NK cell-mediated, anti-HCV activity is supported by the observation that NK cell-conditioned media enhanced expression of signal transducer and activator of transcription-1, a nuclear factor that is essential in IFN-gamma-mediated antiviral pathways. NK cell-conditioned media have the ability to stimulate intracellular IFN-alpha expression in the hepatic cells, suggesting a mechanism responsible for NK cell mediated, anti-HCV activity. Thus, NK cells hold the potential to play a vital role in controlling HCV replication in hepatic cells using an IFN-gamma-dependent mechanism. PMID- 15339941 TI - Cellular composition and ultrastructure of the gill epithelium of larval and adult lampreys: implications for osmoregulation in fresh and seawater. AB - Lampreys, one of the only two surviving groups of agnathan (jawless) vertebrates, contain several anadromous species that, during their life cycle, thus migrate from fresh to seawater and back to freshwater. Lampreys have independently evolved the same overall osmoregulatory mechanisms as the gnathostomatous (jawed) and distantly related teleost fishes. Lamprey gills thus likewise play a central role in taking up and secreting monovalent ions. However, the ultrastructural characteristics and distribution of their epithelial cell types [ammocoete mitochondria-rich (MR) cell, intercalated MR cell, chloride cell and pavement cell] differ in several respects from those of teleosts. The ultrastructural characteristics of these cells are distinctive and closely resemble those of certain ion-transporting epithelia in other vertebrates, for which the function has been determined. The data on each cell type, together with the stage in the life cycle at which it is found, i.e. whether in fresh or seawater, enable the following proposals to be made regarding the ways in which lampreys use their gill epithelial cells for osmoregulating in hypo- and hypertonic environments. In freshwater, the intercalated MR cell takes up Cl- and secretes H+, thereby facilitating the uptake of Na+ through pavement cells. In seawater, the chloride cell uses a secondarily active transcellular transport of Cl- to provide the driving force for the passive movement of Na+ through leaky paracellular pathways between these cells. PMID- 15339942 TI - Test of the mechanotactile hypothesis: neuromast morphology and response dynamics of mechanosensory lateral line primary afferents in the stingray. AB - The mechanotactile hypothesis proposes that ventral non-pored lateral line canals in the stingray function to facilitate localization of prey that contact the skin during benthic feeding. This study used comparative neurophysiological and morphological techniques to test whether ventral non-pored canals encode the velocity of skin movements, and show other adaptations that may enhance detection of tactile stimuli from their prey. Resting discharge rate of lateral line primary afferent neurons was lower among units from ventral than dorsal canal groups. The ventral non-pored canals had a higher proportion of silent units (31%) than either ventral (3%) or dorsal (13%) pored canals, thus may have an enhanced potential for detection of phasic contact with prey. Primary afferents from pored canals showed response characteristics consistent with acceleration detectors, with best frequencies of 20-30 Hz. In contrast, units from non-pored canals responded to tactile skin depression velocities of 30-630 microm s(-1) from 1-20 Hz, and encoded the velocity of canal fluid induced by skin movement with best frequencies of < or =10 Hz. Sensitivity of non-pored canals to direct skin depression velocity was 2-10 times greater than to hydrodynamic dipole stimulation near the skin. No morphological specialization of hair cell orientation was found among pored and non-pored canals. These low frequency, tactile response properties support the hypothesis that the stingray's non-pored ventral lateral line functions as a mechanotactile receptor subsystem used to guide small benthic invertebrates to the ventrally positioned mouth. PMID- 15339943 TI - The role of discontinuous gas exchange in insects: the chthonic hypothesis does not hold water. AB - Insects breathe through valved openings (spiracles) in their cuticle. Many insects open and close their spiracles in a cyclic pattern (discontinuous gas exchange cycles, or DGC). These cycles were observed over half a century ago, their hypothesized function being to minimize loss of water from the tracheal system. However, numerous recent studies have found that respiration accounts for a small fraction of total water loss, and that insects stop performing DGC at times when this pattern would be most useful. Thus, the importance of cyclic gas exchange for water conservation has been challenged. The leading alternative is the chthonic hypothesis, which proposes that DGC originated in insects from hypercapnic (high CO2) environments (e.g. burrows) to aid in release of carbon dioxide. By keeping the spiracles closed, insects would concentrate CO2 and increase the gradient for outward diffusion of CO2. CO2 would be released rapidly when the spiracles opened, and respiratory water loss would be reduced. The chthonic hypothesis therefore predicts that DGC minimizes the ratio of respiratory water loss to CO2 release relative to other modes of gas exchange. We tested the chthonic hypothesis by simultaneously measuring water loss and CO2 release in reproductive females (queens) of the seed-harvester ant Pogonomyrmex barbatus, a burrowing species from North American deserts. Queens used one of three patterns of gas exchange, discontinuous, cyclic and continuous. We resolved the problem of separating cuticular transpiration and respiratory water loss for individuals that used continuous gas exchange by developing a regression method that can be used across all patterns of gas exchange. The ratio of respiratory water loss to CO2 release did not differ among ants using different patterns of gas exchange, in contrast to the expectation of the chthonic hypothesis. Metabolic rate, however, varied with gas-exchange pattern, and was lowest for individuals that used discontinuous gas exchange, intermediate for individuals using cyclic gas exchange, and highest for individuals using continuous gas exchange. PMID- 15339944 TI - The buccal buckle: the functional morphology of venom spitting in cobras. AB - Multiple radiations of Asiatic and African cobras have independently evolved the ability to expel their venom as a pressurized horizontal stream, a behavior commonly referred to as spitting. Though the unique fang morphology of spitting cobras is well known, the functional bases of venom spitting have received little attention. The combined results of gross and microscopic morphology, high-speed digital videography, experimental manipulations of anesthetized cobras and electromyography reveal a two-part mechanism for spitting venom. Contraction of the M. protractor pterygoideus (PP) causes displacement and deformation of the palato-maxillary arch and fang sheath; ultimately this displacement removes soft tissue barriers to venom flow that are normally present within the fang sheath. The M. adductor mandibulae externus superficialis (AMES) is activated simultaneously with the PP; the AMES increases venom pressure within the venom gland, propelling a stream of venom through the venom duct and out the fang. The displacements of the palato-maxillary arch, which form the first part of the spitting mechanism, are very similar to the motions of these bones during prey ingestion (the pterygoid walk), suggesting that venom spitting may have evolved from a specialization of prey ingestion, rather than prey capture. PMID- 15339945 TI - Neuromuscular control of trout swimming in a vortex street: implications for energy economy during the Karman gait. AB - Approximating the complexity of natural locomotor conditions provides insight into the diversity of mechanisms that enable animals to successfully navigate through their environment. When exposed to vortices shed from a cylinder, fishes hold station by adopting a mode of locomotion called the Karman gait, whereby the body of the fish displays large, lateral oscillations and the tail-beat frequency matches the vortex shedding frequency of the cylinder. Although field studies indicate that fishes often prefer turbulent flows over uniform currents, the effect of hydrodynamic perturbations on the mechanics, control and energetics of locomotion is still poorly understood. In this study, electromyography is used to measure red and white axial muscle activity for rainbow trout (Oncorhynchus mykiss) holding station in a vortex street. When trout Karman gait, they show a significantly reduced but still rhythmic pattern of muscle activity compared with that seen when they swim steadily in uniform flow. Specifically, trout selectively activated only their anterior red axial muscles and abandoned the antero-posterior wave of red muscle activity that drives undulatory locomotion in uniform flow. This supports a previously proposed hypothesis that trout are not just swimming in the reduced flow behind a cylinder (drafting). Anterior axial muscle activity was correlated to head amplitude during steady swimming but not during the Karman gait, indicating that while activation of muscles during the Karman gait may aid in stability and control, vortices determined overall head motion. Furthermore, anterior red axial muscle activity, the only region of muscle activity shared between both the Karman gait and steady swimming, had a lower intensity and longer duration during the Karman gait. At times when paired fins were active during the Karman gait, there was no axial muscle activity measured, lending support to a passive mechanism of thrust generation in oscillating flows. Comparisons with dead trout towed behind a cylinder confirm this intriguing observation that live trout may temporarily adopt the Karman gait with no axial muscle activity, revealing paradoxically that at times fish can passively move against turbulent flow. To Karman gait for prolonged periods, however, trout must adapt to the demands of turbulence by eliciting a shift in neural control strategy. By decoupling motor output both down and across the body, the pattern of rhythmic Karman gait muscle activity may reflect the entrainment of a central pattern generator to environmental vortices. PMID- 15339946 TI - Ground reaction forces in horses trotting up an incline and on the level over a range of speeds. AB - Although the forces required to support the body mass are not elevated when moving up an incline, kinematic studies, in vivo tendon and bone studies and kinetic studies suggest there is a shift in forces from the fore- to the hindlimbs in quadrupeds. However, there are no whole-animal kinetic measurements of incline locomotion. Based on previous related research, we hypothesized that there would be a shift in forces to the hindlimb. The present study measured the force produced by the fore- and hindlimbs of horses while trotting over a range of speeds (2.5 to 5 m s(-1)) on both level and up an inclined (10%) surface. On the level, forelimb peak forces increased with trotting speed, but hindlimb peak force remained constant. On the incline, both fore- and hindlimb peak forces increased with speed, but the sum of the peak forces was lower than on the level. On the level, over the range of speeds tested, total force was consistently distributed between the limbs as 57% forelimb and 43% hindlimb, similar to the weight distribution of the horses during static weight tests. On the incline, the force distribution during locomotion shifted to 52% forelimb and 48% hindlimb. Time of contact and duty factor decreased with speed for both limbs. Time of contact was longer for the forelimb than the hindlimb, a finding not previously reported for quadrupeds. Time of contact of both limbs tended to be longer when traveling up the incline than on the level, but duty factor for both limbs was similar under both conditions. Duty factor decreased slightly with increased speed for the hindlimb on the level, and the corresponding small, predicted increase in peak vertical force could not be detected statistically. PMID- 15339947 TI - Distance and force production during jumping in wild-type and mutant Drosophila melanogaster. AB - In many insects renowned for their jumping ability, elastic storage is used so that high forces can be developed prior to jumping. We have combined physiological, behavioural and genetic approaches to test whether elastic energy storage makes a major contribution to jumping in Drosophila. We describe a sensitive strain gauge setup, which measures the forces produced by tethered flies through their mesothoracic legs. The peak force produced by the main jumping muscle of female flies from a wild-type (Canton-S) strain is 101+/-4.4 microN [and this is indistinguishable from a second wild-type (Texas) strain]. The force takes 8.2 ms to reach its peak. The peak force is not affected significantly by altering the leg angle (femur-tibia joint angle) in the range of 75-120 degrees, but the peak force declines as the leg is extended further. Measurements of jumping ability (distance jumped) showed that female Drosophila (with their wings removed) of two wild-type strains, Canton-S and Texas, produced jumps of 28.6+/-0.7 and 30.2+/-1.0 mm (mean +/- s.e.m.). For a female wild-type Drosophila, a jump of 30 mm corresponds to a kinetic energy of 200 nJ on take-off (allowing 20% of the energy to overcome air resistance). We develop equations of motion for a linear force-time model of take-off and calculate that the time to take-off is 5.0 ms and the peak force should be 274 microN (137 microN leg(-1)). We predicted, from the role of octopamine in enhancing muscle tension in several locust muscles, that if stored elastic energy plays no part in force development, then genetic manipulation of the octopaminergic system would directly affect force production and jumping in Drosophila. Using two mutants deficient in the octopaminergic system, TbhnM18 (M18) and TyrRhono (hono), we found significantly reduced jumping distances (20.7+/-0.7 and 20.7+/-0.4 mm, respectively) and force production (52% and 55%, respectively) compared with wild type. From the reduced distance and force production in M18, a mutant deficient in octopamine synthesis, and in hono, a tyramine/octopamine receptor mutant, we conclude that in Drosophila, as in locusts, octopamine modulates escape jumping. We conclude that the fly does not need to store large quantities of elastic energy in order to make its jump because (1) the measured and calculated forces agree to within 40% and (2) the reduction in distances jumped by the mutants correlates well with their reduction in measured peak force. PMID- 15339948 TI - Recognition of calls with exceptionally fast pulse rates: female phonotaxis in the genus Neoconocephalus (Orthoptera: Tettigoniidae). AB - Male Neoconocephalus robustus and Neoconocephalus bivocatus produce remarkably fast calls, with pulse rates of approximately 175-200 Hz. The temporal call patterns differ significantly between the two species. Male N. robustus produce calls with a single pulse rate of 200 Hz. In N. bivocatus, pulses are repeated with alternating periods, resulting in distinct pulse pairs: approximately 175 pulses s(-1) are grouped into 87 pulse pairs s(-1). In order to identify the temporal parameters used to recognize calls with such fast pulse rates, female call recognition in both species was tested during phonotaxis on a walking compensator. Female N. robustus were attracted to calls without amplitude modulation. Amplitude-modulated signals were equally attractive, as long as the silent intervals were short enough. The maximally tolerated interval duration varied with pulse duration. Female N. bivocatus did not require the paired-pulse pattern but were attracted to call models in which each pulse pair was merged into one long pulse. Females used the pulse rate to recognize such signals: pulse rates close to 87 Hz were attractive, largely independent of the duty cycle. Thus, females of the sibling species N. robustus and N. bivocatus used qualitatively different call recognition mechanisms. PMID- 15339949 TI - Environmental split between germ cell parasitism and somatic cell synergism in chimeras of a colonial urochordate. AB - Colonies of the urochordate Botryllus schlosseri may fuse upon contact if they share common alleles on the highly polymorphic fusibility/histocompatibility locus. While, in these chimeras, one of the partners is usually morphologically eliminated (resorbed), circulating totipotent cells of the inferior genotype on the resorption phenomenon may parasitize either the soma or the germ line of the winner. Here, we show an environmental split of the two stem cell lineages that may develop germ cell parasitism vs somatic cell cooperation. Each naturally formed Botryllus chimera can be a composite of component genotypes created through two unlinked parasitic germ and somatic cell lineage interactions. The germ line parasitism is inherited through a pedigree. Conversely, by using amplified fragment length polymorphism (AFLP) and microsatellite alleles as polymorphic genetic markers, and seawater temperature as the variable environmental factor, we documented that the somatic constituent of chimeric zooids was shifted from one genotype to another, in accordance with the changes in seawater temperatures. This variable somatic state of chimerism in the field may, thus, carry benefits to the chimeral entity, which presents synergistically, at any time, the best-fitted combination of its genetic components. PMID- 15339950 TI - Testing olfactory foraging strategies in an Antarctic seabird assemblage. AB - Procellariiform seabirds (petrels, albatrosses and shearwaters) forage over thousands of square kilometres for patchily distributed prey resources. While these birds are known for their large olfactory bulbs and excellent sense of smell, how they use odour cues to locate prey patches in the vast ocean is not well understood. Here, we investigate species-specific responses to 3-methyl pyrazine in a sub-Antarctic species assemblage near South Georgia Island (54 degrees 00 ' S, 36 degrees 00 ' W). Pyrazines are scented compounds found in macerated Antarctic krill (Euphausia superba), a primary prey item for many seabird species in this region. To examine behavioural attraction to this odour, we presented birds with either scented or 'unscented' vegetable oil slicks at sea. As a positive control for our experiments, we also compared birds' responses to a general olfactory attractant, herring oil. Responses to pyrazine were both highly species specific and consistent with results from earlier studies investigating responses to crude krill extracts. For example, Cape petrels (Daption capense), giant petrels (Macronectes sp.) and white-chinned petrels (Procellaria aequinoctialis) were sighted at least 1.8-4 times as often at pyrazine-scented slicks than at control slicks. Black-browed albatrosses (Diomedea melanophris) were only sighted at pyrazine-scented slicks and never at control slicks. Wilson's storm-petrels (Oceanites oceanicus), black-bellied storm petrels (Fregetta tropica), great shearwaters (Puffinus gravis) and prions (Pachyptila sp.) were sighted with equal frequency at control and pyrazine scented slicks. As expected, responses to herring oil were more common. With the exception of great shearwaters (Puffinus gravis), each of these species was sighted up to five times as often at slicks scented with herring oil compared with control slicks. Together, the results support the hypothesis that Antarctic procellariiforms use species-specific foraging strategies that are inter dependent and more complex than simply tracking prey by scent. PMID- 15339951 TI - Biomechanics of quadrupedal walking: how do four-legged animals achieve inverted pendulum-like movements? AB - Walking involves a cyclic exchange of gravitational potential energy and kinetic energy of the center of mass. Our goal was to understand how the limbs of walking quadrupeds coordinate the vertical movements of the fore and hind quarters to produce these inverted pendulum-like movements. We collected kinematic and ground reaction force data from dogs walking over a range of speeds. We found that the fore and hind quarters of dogs behaved like two independent bipeds, each vaulting up and over its respective support limb. The center of mass moved up and down twice per stride, like a single walking biped, and up to 70% of the mechanical energy required to lift and accelerate the center of mass was recovered via the inverted pendulum mechanism. To understand how the limbs produce these center of mass movements, we created a simple model of two independent pendulums representing the movements of the fore and hind quarters. The model predicted that the fore and hind quarter movements would completely offset each other if the fore limb lagged the hind limb by 25% of the stride time and body mass was distributed equally between the fore and hind quarters. The primary reason that dogs did not walk with a flat trajectory of the center of mass was that each fore limb lagged its ipsilateral hind limb by only 15% of the stride time and thereby produced time periods when the fore and hind quarters moved up or down simultaneously. The secondary reason was that the fore limbs supported 63% of body mass. Consistent with these experimental results, the two-pendulum model predicts that the center of mass will undergo two fluctuations per stride cycle if limb phase is less than 25% and/or if the total mass is not distributed evenly between the fore or hind quarters. PMID- 15339952 TI - Plastic and evolved responses of larval tracheae and mass to varying atmospheric oxygen content in Drosophila melanogaster. AB - Structural changes in the tracheal system during development have the potential to allow insects to compensate for varying oxygen availability. Despite possible compensation, oxygen level during development may also affect insect body size. We investigated how atmospheric oxygen level affects the dimensions of the main dorsal tracheae (DT) and masses of larval Drosophila melanogaster (Meigen) reared for up to six generations in 10%, 21% or 40% O2 at 25 degrees C. Wandering-stage third-instar larvae were weighed every other generation, and the dimensions of the DT were measured. Hypoxia produced significantly lighter larvae after one generation of exposure, while hyperoxia did not affect larval mass. Atmospheric oxygen content did not significantly change the diameters of the anterior portions of the main tracheae; however, the posterior diameters were strongly affected. During the first generation of exposure, tracheal diameters were inversely proportional to rearing oxygen levels, demonstrating that developmental plasticity in DT diameters can partially (8-15%) compensate for variation in atmospheric oxygen level. After multiple generations in differing atmospheres and two further generations in 21% O2, larvae had tracheal diameters inversely related to their historical oxygen exposure, suggesting that atmospheric oxygen can produce heritable changes in insect tracheal morphology. PMID- 15339953 TI - Models of hydrodynamic flow in the bowhead whale filter feeding apparatus. AB - Anatomical and behavioral analyses suggest that the filtration mechanism of bowhead and right whales (Balaenidae) is driven by hydrodynamic as well as ram hydraulic pressures. Complementary models were devised to investigate biomechanical aspects of water flow in the buccal cavity of the bowhead whale Balaena mysticetus during continuous filter feeding. A mathematical model was created to test and quantify water flow predictions with steady state hydromechanical equations; a physical model of the bowhead mouth (approximately 1/15 scale) was constructed to visualize flow processes. Both models rely on morphometric data obtained from whales harvested by Inupiat Eskimos for subsistence purposes along with information on foraging ecology (locomotor velocity, gape, etc.). Results indicate that unique features of balaenid oral construction and function (e.g. subrostral gap, orolabial sulcus, curvature of baleen, extensive mandibular rotation and lingual mobility) not only permit steady, unidirectional flow of water through the mouth, but also establish Bernoulli and Venturi effects during feeding. These hydrodynamic conditions allow balaenids to improve filtering efficiency and avoid creation of an anterior compressive wave (by increasing flow velocity and thereby reducing pressure) so that they may capture elusive prey even at slow swimming speeds. PMID- 15339954 TI - Phylogenetic conservation of disulfide-linked, dimeric acetylcholine receptor pentamers in southern ocean electric rays. AB - Intact acetylcholine receptors have been purified on a novel affinity resin from three electric fish endemic to Australian waters. Their binding properties and morphology are compared with those of their northern hemisphere homolog, Torpedo marmorata. All four exhibit apparent dissociation constants, Kd, in the nanomolar range for the snake neurotoxin alpha-bungarotoxin and have a distinctive rosette like appearance when viewed in negative stain under the electron microscope. Furthermore, these rosettes are paired, indicating that acetylcholine receptors from southern ocean electric fish exist as dimers, in the same fashion as their northern hemisphere counterparts. The cDNAs of the receptor's four subunits were sequenced from Hypnos monopterigium and the northern hemisphere counterpart, Torpedo marmorata, while cDNAs from only two subunits, alpha and delta, were able to be sequenced from Narcine tasmaniensis. The penultimate amino acid in the delta subunit of each of the newly sequenced fish species is a cysteine residue. Its conservation suggests that the mechanism for the observed dimerization of acetylcholine receptors is disulfide bond formation between the delta subunit of adjacent receptors, analogous to acetylcholine receptor dimers observed in other electric fish. It appears that this mechanism for receptor clustering is unique to acetylcholine receptors packed and organized in the specialized organs of electric fish. Alignment of the deduced protein sequences with the equivalent sequences from Torpedo californica and humans reveals a high degree of homology. PMID- 15339955 TI - Acoustical stress and hearing sensitivity in fishes: does the linear threshold shift hypothesis hold water? AB - Mammals exposed to loud aerial sounds exhibit temporary threshold shifts (TTS) that are linearly related to increases of sound pressure above baseline hearing levels. It was unknown if this relationship held true for aquatic ectotherms such as fishes. To test this linear threshold shift hypothesis (LINTS) in fishes, we examined the effects of increased ambient sound on hearing of two species differing in hearing capabilities: goldfish (Carassius auratus; a hearing specialist) and tilapia (Oreochromis niloticus; a hearing generalist). Fish were exposed to 1-28 days of either quiet (110 dB re 1 microPa) or continuous white noise. First, we examined the effect of noise sound pressure level (SPL; 130, 140, 160 or 170 dB re 1 microPa) on goldfish hearing thresholds after 24 h of noise exposure. Second, in a long-term experiment using 170 dB re 1 microPa white noise, we continuously exposed goldfish and tilapia for either 7 or 21-28 days. In both experiments, we measured alterations in hearing capabilities (using auditory brainstem responses) of noise-exposed fish. While tilapia exposed to noise for 28 days showed little or no hearing loss, goldfish exhibited considerable threshold shifts that reached an asymptote of up to 25 dB after only 24 h of exposure. There was a positive linear relationship between noise-induced TTS and the sound pressure difference between the noise and the baseline hearing thresholds in goldfish but not in tilapia. A similar relationship was found for published noise-induced threshold shifts in birds and mammals, but the slope of the linear relationship was greater in these groups than for fish. The linear threshold shift relationship provides insights into differential susceptibility of hearing specialist and generalist fishes to noise-induced hearing loss for a given SPL and provides a framework for future research on noise-induced threshold shifts in fishes and other animals. PMID- 15339956 TI - Substantial changes in central nervous system neurotransmitters and neuromodulators accompany phase change in the locust. AB - Desert locusts (Schistocerca gregaria) can undergo a profound transformation between solitarious and gregarious forms, which involves widespread changes in behaviour, physiology and morphology. This phase change is triggered by the presence or absence of other locusts and occurs over a timescale ranging from hours, for some behaviours to change, to generations, for full morphological transformation. The neuro-hormonal mechanisms that drive and accompany phase change in either direction remain unknown. We have used high-performance liquid chromatography (HPLC) to compare amounts of 13 different potential neurotransmitters and/or neuromodulators in the central nervous systems of final instar locust nymphs undergoing phase transition and between long-term solitarious and gregarious adults. Long-term gregarious and solitarious locust nymphs differed in 11 of the 13 substances analysed: eight increased in both the brain and thoracic nerve cord (including glutamate, GABA, dopamine and serotonin), whereas three decreased (acetylcholine, tyramine and citrulline). Adult locusts of both extreme phases were similarly different. Isolating larval gregarious locusts led to rapid changes in seven chemicals equal to or even exceeding the differences seen between long-term solitarious and gregarious animals. Crowding larval solitarious locusts led to rapid changes in six chemicals towards gregarious values within the first 4 h (by which time gregarious behaviours are already being expressed), before returning to nearer long-term solitarious values 24 h later. Serotonin in the thoracic ganglia, however, did not follow this trend, but showed a ninefold increase after a 4 h period of crowding. After crowding solitarious nymphs for a whole larval stadium, the amounts of all chemicals, except octopamine, were similar to those of long term gregarious locusts. Our data show that changes in levels of neuroactive substances are widespread in the central nervous system and reflect the time course of behavioural and physiological phase change. PMID- 15339957 TI - Transmission of linearly polarized light in seawater: implications for polarization signaling. AB - Partially linearly polarized light is abundant in the oceans. The natural light field is partially polarized throughout the photic range, and some objects and animals produce a polarization pattern of their own. Many polarization-sensitive marine animals take advantage of the polarization information, using it for tasks ranging from navigation and finding food to communication. In such tasks, the distance to which the polarization information propagates is of great importance. Using newly designed polarization sensors, we measured the changes in linear polarization underwater as a function of distance from a standard target. In the relatively clear waters surrounding coral reefs, partial (%) polarization decreased exponentially as a function of distance from the target, resulting in a 50% reduction of partial polarization at a distance of 1.25-3 m, depending on water quality. Based on these measurements, we predict that polarization sensitivity will be most useful for short-range (in the order of meters) visual tasks in water and less so for detecting objects, signals, or structures from far away. Navigation and body orientation based on the celestial polarization pattern are predicted to be limited to shallow waters as well, while navigation based on the solar position is possible through a deeper range. PMID- 15339958 TI - Fenretinide activates a distinct apoptotic pathway. PMID- 15339959 TI - Versipelostatin: unfolding an unsweetened death. PMID- 15339960 TI - Medicare cost containment strategy targets several oncology drugs. PMID- 15339961 TI - Three molecularly targeted drugs tested in kidney cancer clinical trials. PMID- 15339962 TI - Stat bite: Incidence of and mortality from kidney cancer, 1975-2001. PMID- 15339963 TI - Recent work adds support to theory that cells may have metastatic origin. PMID- 15339964 TI - CEO Roundtable to reward corporate standards for workplace cancer prevention. PMID- 15339966 TI - Nutrition and physical activity and chronic disease prevention: research strategies and recommendations. AB - A shortage of credible information exists on practical dietary and physical activity patterns that have potential to reverse the national obesity epidemic and reduce the risk of major cancers and other chronic diseases. Securing such information is a challenging task, and there is considerable diversity of opinion concerning related research designs and priorities. Here, we put forward some perspectives on useful methodology and infrastructure developments for progress in this important area, and we list high-priority research topics in the areas of 1) assessment of nutrient intake and energy expenditure; 2) development of intermediate outcome biomarkers; 3) enhancement of cohort and cross-cultural studies; and 4) criteria for and development of full-scale nutrition and physical activity intervention trials. PMID- 15339967 TI - Gangliosides link the acidic sphingomyelinase-mediated induction of ceramide to 12-lipoxygenase-dependent apoptosis of neuroblastoma in response to fenretinide. AB - BACKGROUND: The lipid second messenger ceramide, which is generated by acidic and neutral sphingomyelinases or ceramide synthases, is a common intermediate of many apoptotic pathways. Metabolism of ceramide involves several enzymes, including glucosylceramide synthase and GD3 synthase, and results in the formation of gangliosides (GM3, GD3, and GT3), which in turn promote the generation of reactive oxygen species (ROS) and apoptosis. Fenretinide, a retinoic acid derivative, is thought to induce apoptosis via increases in ceramide levels, but the link between ceramide and subsequent apoptosis in neuroblastoma cells is unclear. METHODS: SH-SY5Y and HTLA230 neuroblastoma cells were treated with fenretinide in the presence or absence of inhibitors of enzymes important in ceramide metabolism (fumonisin B1, inhibitor of ceramide synthase; desipramine, inhibitor of acidic and neutral sphingomyelinases; and PDMP, inhibitor of glucosylceramide). Small interfering RNAs were used to specifically block acidic sphingomyelinase or GD3 synthase activities. Apoptosis, ROS, and GD3 expression were measured by flow cytometry. RESULTS: In neuroblastoma cells, ROS generation and apoptosis were associated with fenretinide-induced increased levels of ceramide, glucosylceramide synthase activity, GD3 synthase activity, and GD3. Fenretinide also induced increased levels of GD2, a ganglioside derived from GD3. Inhibition of acidic sphingomyelinase but not of neutral sphingomyelinase or ceramide synthase, blocked fenretinide-induced increases in ceramide, ROS, and apoptosis. Exogenous GD3 induced ROS and apoptosis in SH-SY5Y cells but not in SH SY5Y cells treated with baicalein, a specific 12-lipoxygenase inhibitor. Exogenous GD2 did not induce apoptosis. CONCLUSIONS: A novel pathway of fenretinide-induced apoptosis is mediated by acidic sphingomyelinase, glucosylceramide synthase, and GD3 synthase, which may represent targets for future drug development. GD3 may be a key signaling intermediate leading to apoptosis via the activation of 12-lipoxygenase. PMID- 15339968 TI - Effect on tumor cells of blocking survival response to glucose deprivation. AB - BACKGROUND: Glucose deprivation, a feature of poorly vascularized solid tumors, activates the unfolded protein response (UPR), a stress-signaling pathway, in tumor cells. We recently isolated a novel macrocyclic compound, versipelostatin (VST), that inhibits transcription from the promoter of GRP78, a gene that is activated as part of the UPR. We examined the effect of VST on the UPR induced by glucose deprivation or other stressors and on tumor growth in vivo. METHODS: Human colon cancer HT-29, fibrosarcoma HT1080, and stomach cancer MKN74 cells were cultured in the absence of glucose or in the presence of glucose and a UPR inducing chemical stressor (the N-glycosylation inhibitor tunicamycin, the calcium ionophore A23187, or the hypoglycemia-mimicking agent 2-deoxyglucose [2DG]). The effect of VST on UPR induction was determined by reverse transcription-polymerase chain reaction and immunoblot analysis of the UPR target genes GRP78 and GRP94; by immunoblot analysis of the UPR transcriptional activators ATF6, XBP1, and ATF4; and by analyzing reporter gene expression in cells transiently transfected with a GRP78 promoter-reporter gene. Cell sensitivity to VST was examined with a colony formation assay and flow cytometry. In vivo antitumor activity of VST was assessed with an MKN74 xenograft model. RESULTS: VST inhibited expression of UPR target genes in glucose-deprived or 2DG treated cells but not in cells treated with tunicamycin or A23187. VST also inhibited the production of the UPR transcriptional activators XBP1 and ATF4 during glucose deprivation. The UPR-inhibitory action of VST was seen only in conditions of glucose deprivation and caused selective and massive killing of the glucose-deprived cells. VST alone and in combination with cisplatin statistically significantly (P =.004 and P<.001 for comparisons with untreated control, respectively) inhibited tumor growth of MKN74 xenografts. CONCLUSION: Disruption of the UPR may provide a novel therapeutic approach to targeting glucose-deprived solid tumors. PMID- 15339969 TI - Probabilities of death from breast cancer and other causes among female breast cancer patients. AB - BACKGROUND: Among cancer patients, probabilities of death from that cancer and other causes in the presence of competing risks are optimal measures of prognosis and of mortality across demographic groups. We used data on breast cancer patients from the Surveillance, Epidemiology, and End Results (SEER) Program in a competing-risk analysis. METHODS: We determined vital status and cause of death for 395,251 white and 35,259 black female patients with breast cancer diagnosed from January 1, 1973, through December 31, 2000, by use of SEER data. We calculated probabilities of death from breast cancer and other causes according to stage, race, and age at diagnosis; for cases diagnosed from January 1, 1990, to December 31, 2000, we also calculated some such probabilities according to tumor size and estrogen receptor (ER) status. All statistical tests were two sided. RESULTS: The probability of death from breast cancer after nearly 28 years of follow-up ranged from 0.03 to 0.10 for patients with in situ disease to 0.70 to 0.85 for patients with distant disease, depending on race and age. The probability of death from breast cancer at the end of the follow-up period generally declined with age at diagnosis; the probability among the oldest (> or =70 years) compared with the youngest (<50 years) patients was 33% lower for white and 46% lower for black patients with localized disease and 14% lower for white patients and 13% lower for black patients with distant disease. The probability of death from breast cancer exceeded that from all other causes for patients diagnosed with localized disease before age 50 years, with regional disease before age 60 years, and with distant disease at any age. The probability of death from breast cancer for patients diagnosed with localized or regional disease was statistically significantly greater in black patients than in white patients (all six P values < or =.01 for age groups 30-49 to 60-69 years; two P values < or =.04 for ages > or =70 years). Among patients with localized or regional disease and known ER status, the probability of death from breast cancer after nearly 11 years of follow-up ranged from 0.04 to 0.11 for patients with localized ER-positive tumors of 2 cm or less to 0.37 to 0.53 for patients with regional ER-negative tumors. CONCLUSIONS: The probability of death from breast cancer versus other causes varied substantially according to stage, tumor size, ER status, and age at diagnosis in both white and black patients. PMID- 15339970 TI - Burden of illness in cancer survivors: findings from a population-based national sample. AB - BACKGROUND: Population trends in aging and improved cancer survival are likely to result in increased cancer prevalence in the United States, but few estimates of the burden of illness among cancer survivors are currently available. The purpose of this study was to estimate the burden of illness in cancer survivors in a national, population-based sample. METHODS: A total of 1823 cancer survivors and 5469 age-, sex-, and educational attainment-matched control subjects were identified from the 2000 National Health Interview Survey. Multiple measures of burden, including utility, a summary measure of health, and days lost from work, were compared using two-sided tests of statistical significance for the two groups overall and for subgroups stratified by tumor site and time since diagnosis. RESULTS: Compared with matched control subjects, cancer survivors had poorer outcomes across all burden measures (P<.01). Cancer survivors had lower utility values (0.74 versus 0.80; P<.001) and higher levels of lost productivity and were more likely to report their health as fair or poor (31.0% versus 17.9%; P<.001) than matched control subjects. Cancer survivors reported statistically significantly higher burden than did control subjects across tumor sites and across time since diagnosis (i.e., within the past year, 2-5 years, 6-10 years, and > or =11 years for the majority of measures. CONCLUSIONS: Cancer survivors have poorer health outcomes than do similar individuals without cancer across multiple burden measures. These decrements are consistent across tumor sites and are found in patients many years following reported diagnosis. Improved measurement of long-term burden of illness will be important for future prospective research. PMID- 15339971 TI - Multiparametric flow cytometric analysis of inter-patient variation in STAT1 phosphorylation following interferon Alfa immunotherapy. AB - BACKGROUND: Regulation of gene expression by signal transducer and activator of transcription 1 (STAT1) within host tissues mediates the antitumor effects of interferon alfa (IFN alpha). We used a novel flow cytometric assay to examine phosphorylation-mediated activation of STAT1 within immune effector cell subsets following in vitro or in vivo IFN alpha treatments. METHODS: Peripheral blood mononuclear cells (PBMCs) isolated from healthy donors (n = 17) or melanoma patients (n = 19) were treated in vitro with interferon alfa-2b (IFN alpha-2b) or phosphate-buffered saline (PBS) and subjected to multiparametric flow cytometry to measure the levels of phosphorylated STAT1 (P-STAT1) within immune cell subsets. We similarly analyzed PBMCs isolated from melanoma patients before and 1 hour after immunotherapy with IFN alpha-2b. All statistical tests were two-sided. RESULTS: P-STAT1 levels in all major immune cell subsets increased within 15 minutes of in vitro IFN alpha-2b treatment of PBMCs; the increase was most pronounced in T lymphocytes and monocytes. Relatively low doses of IFN alpha-2b (i.e., 10(2)-10(3) IU/mL) induced maximal STAT1 activation in vitro. Compared with melanoma patients, healthy donors had higher basal levels of P-STAT1 (specific fluorescence [Fsp]; i.e., Fsp(PBS), the level of P-STAT1 in PBS-treated cells) in total PBMCs, natural killer (NK) cells, and T cells (mean Fsp(PBS) in total PBMCs: 5.5 in healthy donors versus 1.6 in patients, difference = 3.9, 95% confidence interval [CI] = 1.4 to 6.5, P =.004; mean Fsp(PBS) in NK cells: 4.6 in healthy donors versus 0.9 in patients, difference = 3.7, 95% CI = 1.7 to 5.7, P =.001; mean Fsp(PBS) in T cells: 6.8 in healthy donors versus 0.9 in patients, difference = 5.9, 95% CI = 2.5 to 9.3, P =.002). P-STAT1 was detected in the NK and T cells of two patients who received IFN alpha-2b immunotherapy (20 MU/m2 [MU = million units], administered by intravenous injection). P-STAT1 levels in the PBMCs of a patient treated sequentially with 5 MU/m2 and 10 MU/m2 IFN alpha-2b (administered by subcutaneous injection) also increased in response to treatments with IFN alpha-2b but did not increase further with the increased dosage of IFN alpha-2b. CONCLUSION: This flow cytometry method can be used to monitor STAT1 activation within subsets of immune cells from patients undergoing IFN alpha immunotherapy. PMID- 15339973 TI - Danger signaling by glomerular podocytes defines a novel function of inducible B7 1 in the pathogenesis of nephrotic syndrome. PMID- 15339974 TI - Inhibitors of HMG-CoA reductase reduce receptor-mediated endocytosis in human kidney proximal tubular cells. AB - The proximal tubular cells of the kidney are responsible for reabsorption of proteins from the tubular lumen. In a study using Opossum kidney (OK) cells, receptor-mediated protein endocytosis was reduced by statins, inhibitors of 3 hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase, which are widely used for therapeutic reduction of plasma cholesterol levels. To explore the possible clinical relevance of the observations in OK cells, protein endocytosis in human kidney tubular cells was investigated in the presence and absence of statins. The uptake of FITC-labeled albumin in these cultures of human kidney tubular cells was investigated by microscopy, flow cytometry and spectrofluorometry. Protein uptake occurred selectively into proximal tubular cells while it was absent in distal tubular/collecting duct cells. Three statins (simvastatin, pravastatin, and rosuvastatin) significantly inhibited the uptake of protein in a concentration-dependent way. This inhibitory effect of statins could be prevented by the co-addition of mevalonate, the product of HMG-CoA reductase. This effect was not the result of a statin-induced cytotoxicity since cell-viability was unaffected. Finally, it was demonstrated that statins strongly inhibited cholesterol synthesis in the human kidney tubular cells. These data suggest that statins have the potential to inhibit albumin uptake by the human proximal nephron as a result of inhibition of HMG-CoA reductase in the proximal tubule cells. Taken into account the data of the accompanying manuscript this inhibitory effect most probably results from a reduced prenylation of some proteins critically involved in endocytosis. It is suggested that these data help to explain the occurrence of proteinuria in some patients treated with high statin doses. PMID- 15339975 TI - Inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase reduce receptor-mediated endocytosis in opossum kidney cells. AB - Renal proximal tubule cells are responsible for the reabsorption of proteins that are present in the tubular lumen. This occurs by receptor-mediated endocytosis, a process that has a requirement for some GTP-binding proteins. Statins are inhibitors of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase used for the therapeutic reduction of cholesterol-containing plasma lipoproteins. However, they can also reduce intracellular levels of isoprenoid pyrophosphates that are derived from the product of the enzyme, mevalonate, and are required for the prenylation and normal function of GTP-binding proteins. The hypothesis that inhibition of HMG-CoA reductase in renal proximal tubule cells could reduce receptor mediated-endocytosis was therefore tested. Five different statins inhibited the uptake of FITC-labeled albumin by the proximal tubule-derived opossum kidney cell line in a dose-dependent manner and in the absence of cytotoxicity. The reduction in albumin uptake was related to the degree of inhibition of HMG-CoA reductase. Simvastatin (e.g., statin) inhibited receptor mediated endocytosis of both FITC-albumin and FITC-beta(2)-microglobulin to similar extents but without altering the binding of albumin to the cell surface. The effect on albumin endocytosis was prevented by mevalonate and by the isoprenoid geranylgeranyl pyrophosphate but not by cholesterol. Finally, evidence that the inhibitory effect of statins on endocytosis of proteins may be caused by reduced prenylation and thereby decreased function of one or more GTP-binding proteins is provided. These data establish the possibility in principle that inhibition of HMG-CoA reductase by statins in proximal tubule cells may reduce tubular protein reabsorption. PMID- 15339976 TI - Increased endothelin activity mediates augmented distal nephron acidification induced by dietary protein. AB - The hypothesis that increased dietary protein augments distal nephron acidification and does so through an endothelin (ET-1)-dependent mechanism was tested. Munich-Wistar rats that ate minimum electrolyte diets of 50% (HiPro) and 20% (CON) casein-provided protein, the latter comparable to standard diet, were compared. HiPro versus CON had higher distal nephron net HCO(3) reabsorption by in vivo microperfusion (37.8 +/- 3.2 versus 16.6 +/- 1.5 pmol/mm per min; P < 0.001) as a result of higher H(+) secretion (41.3 +/- 4.0 versus 23.0 +/- 2.1 pmol/mm per min; P < 0.002) and lower HCO(3) secretion (-3.5 +/- 0.4 versus -6.4 +/- 0.8 pmol/mm per min; P < 0.001). Perfusion with H(+) inhibitors support that increased H(+) secretion was mediated by augmented Na(+)/H(+) exchange and H(+) ATPase activity without augmented H(+),K(+)-ATPase activity. HiPro versus CON had higher levels of urine ET-1 excretion, renal cortical ET-1 addition to microdialysate in vivo, and renal cortical ET-1 mRNA, consistent with increased renal ET-1 production. Oral bosentan, an ET A/B receptor antagonist, decreased distal nephron net HCO(3) reabsorption (22.4 +/- 1.9 versus 37.8 +/- 3.2 pmol/mm per min; P < 0.001) as a result of lower H(+) secretion (28.4 +/- 2.4 versus 41.3 +/- 4.0 pmol/mm per min; P < 0.016) and higher HCO(3) secretion (-6.0 +/- 0.7 versus -3.5 +/- 0.4 pmol/mm per min; P < 0.006). The H(+) inhibitors had no additional effect in HiPro ingesting bosentan, supporting that ET mediated the increased distal nephron Na(+)/H(+) exchange and H(+)-ATPase activity in HiPro. Increased dietary protein augments distal nephron acidification that is mediated through an ET-sensitive increase in Na(+)/H(+) exchange and H(+)-ATPase activity. PMID- 15339977 TI - Altered renal distal tubule structure and renal Na(+) and Ca(2+) handling in a mouse model for Gitelman's syndrome. AB - Gitelman's syndrome, an autosomal recessive renal tubulopathy caused by loss-of function mutations in the thiazide-sensitive NaCl co-transporter (NCC) of the distal convoluted tubule (DCT), is characterized by mild renal Na(+) wasting, hypocalciuria, hypomagnesemia, and hypokalemic alkalosis. For gaining further insights into the pathophysiology of Gitelman's syndrome, the impact of NCC ablation on the morphology of the distal tubule, on the distribution and abundance of ion transport proteins along its length, and on renal tubular Na(+) and Ca(2+) handling in a gene-targeted mouse model was studied. NCC-deficient mice had significantly elevated plasma aldosterone levels and exhibited hypocalciuria, hypomagnesemia, and compensated alkalosis. Immunofluorescent detection of distal tubule marker proteins and ultrastructural analysis revealed that the early DCT, which physiologically lacks epithelial Na(+) (ENaC) and Ca(2+) (TRPV5) channels, was virtually absent in NCC-deficient mice. In contrast, the late DCT seemed intact and retained expression of the apical ENaC and TRPV5 as well as basolateral Na(+)-Ca(2+) exchanger. The connecting tubule exhibited a marked epithelial hypertrophy accompanied by an increased apical abundance of ENaC. Ca(2+) reabsorption seemed unaltered in the distal convolution (i.e., the DCT and connecting tubule) as indicated by real-time reverse transcription-PCR, Western blotting, and immunohistochemistry for TRPV5 and Na(+)-Ca(2+) exchanger and micropuncture experiments. The last experiments further indicated that reduced glomerular filtration and enhanced fractional reabsorption of Na(+) and Ca(2+) upstream and of Na(+) downstream of the DCT provide some compensation for the Na(+) transport defect in the DCT and contribute to the hypocalciuria. Thus, loss of NCC leads to major structural remodeling of the renal distal tubule that goes along with marked changes in glomerular and tubular function, which may explain some of the clinical features of Gitelman's syndrome. PMID- 15339978 TI - Podocalyxin activates RhoA and induces actin reorganization through NHERF1 and Ezrin in MDCK cells. AB - Podocalyxin (PC) is the major sialoglycoprotein expressed on the apical membrane of the podocyte. Previously it was shown that PC is connected to actin through the PC/NHERF2/ezrin complex, and this connection is disrupted in the nephrotic syndrome. For assessing whether expression of PC affects the organization of the actin cytoskeleton, MDCK cell lines stably expressing either full-length PC or a PC mutant lacking the NHERF binding site was established. It was found that full length PC but not the PC mutant is connected to actin, induces redistribution of actin toward the apical membrane, and leads to increased RhoA activity. By immunofluorescence redistribution of RhoA and RhoGDI was observed in the presence of both full-length PC and the PC mutant. With the use of pulldown assays, PC and ezrin were found to interact directly and the ezrin binding site was mapped to the juxtamembrane region of PC's cytoplasmic tail. It is concluded that PC binds to ezrin both directly and indirectly. PC activates RhoA through NHERF and ezrin, leading to redistribution of actin filaments. These results suggest that in podocytes, PC may also regulate foot process architecture through RhoA. PMID- 15339979 TI - Growth hormone-mediated janus associated kinase-signal transducers and activators of transcription signaling in the growth hormone-resistant potassium-deficient rat. AB - Potassium deficiency (KD) is associated with severe growth failure, in part caused by growth hormone (GH) resistance. This study set out to determine whether the resistance could be caused by a defect in GH-mediated janus associated kinase signal transducers and activators of transcription (STAT) signaling as occurs in uremia. To this end, rats were fed a K-deficient diet for 8 d and pair-fed controls received a K-replete diet. Animals from each group received GH or vehicle, and during this period, KD rats were GH resistant; GH induced body and liver weight gain and linear body growth were severely attenuated in these rats. In addition, signal transduction was studied in the liver of rats that were killed 10 or 15 min after an intravenous GH bolus or vehicle. When the rats were killed, GH receptor mRNA and protein levels were similar in the two groups. The abundance of STAT5, STAT3, and STAT1, proteins that mediate GH signaling, was significantly increased by 40 to 130% in KD. Furthermore, GH induced a far greater increase in STAT5 and STAT3 phosphorylation in this group. STAT5 phosphorylation was enhanced fourfold even when normalized for total STAT5 content. Phosphorylated STAT5 and STAT3 proteins were also increased in nuclear extracts, suggesting normal nuclear translocation of the activated signaling proteins. DNA binding of nuclear STAT5 was unaltered. Thus, in KD, there is resistance to the growth-promoting action of GH despite hyperactivation of the janus associated kinase-STAT signaling pathway. This suggests the presence of a defect distal to the nuclear binding of STAT or, alternatively, a defect in a STAT-independent GH-activated signaling pathway. PMID- 15339980 TI - Inducible nitric oxide synthase-derived nitric oxide promotes glomerular angiogenesis via upregulation of vascular endothelial growth factor receptors. AB - The vascular endothelial growth factor (VEGF) system is of major importance for glomerular endothelial repair in glomerulonephritis (GN) and is significantly affected by nitric oxide (NO) release. For investigating whether glomerular upregulation of inducible NO synthase (iNOS) in GN might affect VEGF-mediated repair, a selective iNOS inhibitor, L-N6-(1-iminoethyl)-lysin (L-NIL), was administered to rats with anti-Thy 1.1 GN from day -2 until day 5 after GN induction. Compared with untreated nephritic rats, L-NIL-treated nephritic rats showed similar mean arterial BP, significantly decreased de novo peak nitrate production, and increased albuminuria on day 6. This was preceded by a significant decrease of glomerular endothelial cell proliferation and endothelial area on day 2 in L-NIL-treated nephritic rats. Upregulation of glomerular VEGF mRNA and protein expression, in particular of the VEGF(164) splicing variant, occurred similarly in L-NIL-treated and untreated nephritic rats on days 2 and 7. However, the upregulation of glomerular VEGF receptor 1 and 2 mRNA expression on day 2 was reduced by 77 and 67%, respectively, in L-NIL-treated nephritic rats as compared with untreated nephritic rats. In parallel, glomerular VEGF(165) binding was reduced by 34% in L-NIL-treated nephritic rats on day 2. Glomerular upregulation of the VEGF(164) co-receptor neuropilin-1 mRNA in nephritic rats was reduced by L-NIL treatment only on day 7. Healthy untreated or L-NIL-treated controls showed no significant differences in any parameter analyzed. In conclusion, impaired repair of glomerular endothelium and downregulation of glomerular VEGF receptor expression was observed after selective iNOS inhibition in experimental GN. These data identify iNOS-derived NO production as the first in vivo regulator of the glomerular VEGF system and as an important mechanism promoting glomerular healing. PMID- 15339981 TI - Blockade of calcium influx through L-type calcium channels attenuates mitochondrial injury and apoptosis in hypoxic renal tubular cells. AB - In hypoxia, ATP depletion causes cellular Ca(2+) increase, mitochondrial injury, and apoptosis in renal tubular cells. However, the molecular basis of these observations is incompletely delineated. IRPTC, a rat renal proximal tubular cell line, was treated with antimycin A, and disturbances in cytoplasmic calcium ([Ca(2+)]c) and mitochondrial calcium ion concentration ([Ca(2+)]m), dissipation of mitochondrial membrane potential (DeltaPsi(m)), cytochrome c release, and resultant apoptosis were examined. Pharmacologic targeting of L-type Ca(2+) channels in vitro and in vivo was used to clarify the involvement of voltage dependent Ca(2+) channels during this process. In vitro studies indicated that ATP depletion-induced apoptosis was preceded by increased [Ca(2+)]c and [Ca(2+)]m before activation of mitochondrial signaling. Antagonizing L-type Ca(2+) channels offset these findings, suggesting [Ca(2+)]c and [Ca(2+)]m involvement. Azelnidipine administration ameliorated cellular and mitochondrial Ca(2+) accumulation, mitochondrial permeability transition, cytochrome c release, caspase-9 activation, and resultant apoptosis (15.8 +/- 0.8% versus 8.9 +/- 0.7%; P < 0.01). Similar effects of azelnidipine were substantiated in an in vivo ischemia/reperfusion injury model. There were fewer terminal-deoxynucleotidyl transferase mediated dUTP nick-end labeling-positive cells in the azelnidipine treated group (0.322 +/- 0.038/tubule) as compared with the vehicle-treated group (0.450 +/- 0.041; P < 0.05), although the antiapoptotic effect was smaller in vivo than in vitro, partly as a result of distinct levels of Bax expression. It is proposed that voltage-dependent Ca(2+) channels are involved in cellular and mitochondrial accumulation of Ca(2+) subsequent to ATP depletion and play an important role in regulating mitochondrial permeability transition, cytochrome c release, caspase activation, and apoptosis. PMID- 15339982 TI - Verotoxin (shiga toxin) sensitizes renal epithelial cells to increased heme toxicity: possible implications for the hemolytic uremic syndrome. AB - Escherichia coli-derived verotoxins (VT; Shiga toxins) are causally related to the pathogenesis of enteropathic hemolytic uremic syndrome (HUS). Profound hemolysis is a defining feature of the disease, but it is not known whether the acute intravascular release of heme proteins contributes to HUS pathology. This study examined the biologic effects of hemin and VT by means of tubular epithelial-derived ACHN cells. Hemin at concentrations >/=200 microM caused cell rounding, spike formation, and detachment that was morphologically distinct from verocytotoxicity. VT caused apoptosis at concentrations >100 pM, as demonstrated by nuclear segmentation and poly(ADP-ribose) polymerase cleavage, whereas hemin mediated injury of ACHN cells grown in serum-containing medium lacked attributes of programmed cell death. Pretreatment of ACHN monolayers with sublethal concentrations (1 to 10 pM) of VT for 12 to 18 h led to superadditive hemin mediated cytotoxicity. This effect was not limited to ACHN cells, but was similarly noted in microvascular endothelial cells. Heme catabolism is regulated by (inducible) heme oxygenase-1 (HO-1). VT abrogated HO-1 expression in ACHN cells. Stimulation of cells for 6 h with CdCl(2), which markedly increased HO-1 expression before the addition of VT, blunted subsequent hemin injury. In conclusion, VT augments hemin-induced toxicity in renal tubular epithelial cells that can be reversed by prior induction of HO-1. It is proposed that VT subverts the physiologic defense against heme proteins by interfering with the regulated expression of HO-1 and that this mechanism contributes to the renal pathology in patients with Escherichia coli-associated HUS. PMID- 15339983 TI - Identifying the molecular phenotype of renal progenitor cells. AB - Although many of the molecular interactions in kidney development are now well understood, the molecules involved in the specification of the metanephric mesenchyme from surrounding intermediate mesoderm and, hence, the formation of the renal progenitor population are poorly characterized. In this study, cDNA microarrays were used to identify genes enriched in the murine embryonic day 10.5 (E10.5) uninduced metanephric mesenchyme, the renal progenitor population, in comparison with more rostral derivatives of the intermediate mesoderm. Microarray data were analyzed using R statistical software to determine accurately genes differentially expressed between these populations. Microarray outliers were biologically verified, and the spatial expression pattern of these genes at E10.5 and subsequent stages of early kidney development was determined by RNA in situ hybridization. This approach identified 21 genes preferentially expressed by the E10.5 metanephric mesenchyme, including Ewing sarcoma homolog, 14-3-3 theta, retinoic acid receptor-alpha, stearoyl-CoA desaturase 2, CD24, and cadherin-11, that may be important in formation of renal progenitor cells. Cell surface proteins such as CD24 and cadherin-11 that were strongly and specifically expressed in the uninduced metanephric mesenchyme and mark the renal progenitor population may prove useful in the purification of renal progenitor cells by FACS. These findings may assist in the isolation and characterization of potential renal stem cells for use in cellular therapies for kidney disease. PMID- 15339984 TI - Thromboxane receptor mediates renal vasoconstriction and contributes to acute renal failure in endotoxemic mice. AB - Sepsis is a major cause of acute renal failure (ARF) and death. Thromboxane A2 (TxA(2)) may mediate decreases of renal blood flow (RBF) and/or GFR associated with LPS-induced sepsis. This study tested whether TxA(2) receptor blockade, with the use of TxA(2) receptor knockout (TP-KO) mice or a selective TP receptor antagonist (SQ29,548), would alleviate LPS-induced renal vasoconstriction and ARF. Under basal conditions, anesthetized TP-KO mice displayed a lower mean arterial pressure than wild-type (WT) mice (102 versus 94 mmHg; P < 0.05). RBF, renal vascular resistance (RVR), GFR, and urine flow did not differ among groups under basal conditions, suggesting little tonic influence of TxA(2) on renal TP receptors in health. In endotoxemic WT mice, 14 h after LPS (Escherichia coli LPS 8.5 mg/kg intraperitoneally), mean arterial pressure was reduced to 85 mmHg (P < 0.001), as were RBF (5.0 versus 9.3 ml/min per g kidney wt; P < 0.001) and GFR (0.38 versus 1.03 ml/min per g kidney wt; P < 0.001). Heart rate and RVR (71 versus 47 mmHg/ml per min; P < 0.05) increased. The decreases in RBF and GFR after LPS were attenuated in TP-KO mice versus WT mice (both P < 0.05). In both TP-KO and TP antagonist-treated mice, RVR remained stable in response to LPS versus WT mice that did not receive LPS. Delayed TP-antagonist treatment (12 h after LPS injection) ameliorated RBF and RVR but did not restore GFR. In other WT animals, TP-antagonist treatment for 2 h before intravenous LPS abolished the early renal vasoconstriction and alleviated the decrease in GFR. These results demonstrate that renal vasoconstriction during endotoxemic shock induced by LPS is mediated by TP receptors as indicated by pharmacologic blockade and genetic disruption of TP receptors. PMID- 15339985 TI - Leptin as a uremic toxin interferes with neutrophil chemotaxis. AB - Leptin is a pleiotropic molecule involved in energy homeostasis, hematopoiesis, inflammation, and immunity. Hypoleptinemia characterizing starvation has been strictly related to increased susceptibility to infection secondary to malnutrition. Nevertheless, ESRD is characterized by high susceptibility to bacterial infection despite hyperleptinemia. Defects in neutrophils play a crucial role in the infectious morbidity, and several uremic toxins that are capable of depressing neutrophil functions have been identified. Only a few and contrasting reports about leptin and neutrophils are available. This study provides evidence that leptin inhibits neutrophil migration in response to classical chemoattractants. Moreover, serum from patients with ESRD inhibits migration of normal neutrophils in response to N-formyl-methionyl-leucyl phenylalanine with a strict correlation between serum leptin levels and serum ability to suppress neutrophil locomotion. Finally, the serum inhibitory activity can be effectively prevented by immune depletion of leptin. The results also show, however, that leptin by itself is endowed with chemotactic activity toward neutrophils. The two activities-inhibition of the cell response to chemokines and stimulation of neutrophil migration-could be detected at similar concentrations. On the contrary, neutrophils exposed to leptin did not display detectable [Ca(2+)](i) mobilization, oxidant production, or beta(2)-integrin upregulation. The results demonstrate that leptin is a pure chemoattractant devoid of secretagogue properties that are capable of inhibiting neutrophil chemotaxis to classical neutrophilic chemoattractants. Taking into account the crucial role of neutrophils in host defense, the leptin-mediated ability of ERSD serum to inhibit neutrophil chemotaxis appears as a potential mechanism that contributes to the establishment of infections in ERSD. PMID- 15339986 TI - Endogenous IL-13 limits humoral responses and injury in experimental glomerulonephritis but does not regulate Th1 cell-mediated crescentic glomerulonephritis. AB - IL-13 is produced by T helper 2 (Th2) cells, has a role in stimulating Th2 mediated injury, alters humoral responses, and may directly suppress macrophage and neutrophil function. In immune renal disease, the engagement of different effector mediator systems, including humoral and cell-mediated effectors, can result in glomerular injury. Experimental crescentic glomerulonephritis (known as autologous anti-glomerular basement membrane glomerulonephritis) induced by planting an antigen in glomeruli of mice is Th1 directed, delayed-type hypersensitivity (DTH)-like, and antibody independent. To test the hypothesis that, like the counterregulatory Th2 cytokines IL-4 and IL-10, endogenous IL-13 limits effector Th1 responses in glomerulonephritis, crescentic glomerulonephritis was induced in IL-13+/+ and IL-13-/- mice. Although IL-13-/- mice developed increased serum antigen-specific antibody levels, increased glomerular antibody deposition and enhanced switching to the Th1-associated IgG2a subclass, they developed a similar degree of crescentic glomerulonephritis, with similar glomerular T cell/macrophage numbers, renal impairment, and proteinuria. Antigen-specific dermal DTH and IFN-gamma production by antigen-stimulated splenocytes was unaltered. In immune complex (apoferritin-induced) glomerulonephritis, where renal injury is humorally mediated, IL-13-/- mice developed enhanced humoral immune responses and increased proteinuria, with increased IgG2a responses, a more peripheral distribution of immune complexes, but no alterations in leukocyte recruitment. These results demonstrate dissociation of IL-13's effects in antigen induced renal disease with little effect on cellular responses but suppressive effects on humoral effectors and switching to IgG2a. They indicate a role for IL-13 in limiting antibody-mediated renal injury, but not in regulating DTH-like cell-mediated responses in the kidney. PMID- 15339987 TI - CCAAT/Enhancer-binding protein delta contributes to myofibroblast transdifferentiation and renal disease progression. AB - Myofibroblasts are pivotal participants in pathologic processes in a wide variety of organs, such as lung, liver, and kidney, by producing several inflammatory cytokines and extracellular matrices. The mechanism by which transdifferentiation from original cell to myofibroblast occurs, however, is still unclear. The expression of smooth muscle alpha-actin (SMalphaA) is the most characteristic feature of myofibroblasts; therefore, it was speculated that any factors that promote SMalphaA expression might be the key to transdifferentiation to myofibroblasts and disease exacerbation. A transcription factor CCAAT/enhancer binding protein delta (C/EBPdelta) was identified and demonstrated to bind to sequences including the CArG motif from SMalphaA intron 1 and to increase transcriptional activity of this promoter. Expression of SMalphaA and C/EBPdelta in the glomerular area was upregulated in rat anti-Thy1 glomerulonephritis and mouse Habu-venom glomerulonephritis, both of which are models of mesangioproliferative glomerulonephritis. In the latter model, C/EBPdelta knockout mice demonstrated significantly less SMalphaA expression in the glomerular area on day 8 and less renal functional deterioration on day 14, compared with wild-type mice. These data suggest an important role for C/EBPdelta in myofibroblast transdifferentiation and glomerulonephritis exacerbation. PMID- 15339988 TI - Nephropathy in Zucker diabetic fat rat is associated with oxidative and nitrosative stress: prevention by chronic therapy with a peroxynitrite scavenger ebselen. AB - Zucker diabetic fat (ZDF) rats with the metabolic syndrome and hyperlipidemia develop focal and segmental sclerosis. The role of oxidative and nitrosative stress in the nephropathy in ZDF was studied. Renal histology, function, and immunohistologic and biochemical parameters of oxidative and nitrosative stress were evaluated at 8 and 22 wk of age in ZDF and Zucker lean (ZL) rats and after chronic treatment with ebselen, an antioxidant and peroxinitrite scavenger. At 8 wk, ZDF rats showed hyperglycemia, no proteinuria or nephropathy, but higher levels of dihydrobiopterin and 3-nitrotyrosine (3-NT)-modified proteins compared with age-matched ZL rats. At 22 wk, ZDF rats developed focal and segmental sclerosis, proteinuria, decreased creatinine clearance, and renal tissue levels of glutathione and tetrahydrobiopterin with further elevation in dihydrobiopterin and 3-NT-modified proteins, in contrast to age-matched ZL rats. Renal immunohistologic expression of lipid peroxidation products and 3-NT-modified proteins also increased in 22-wk-old ZDF but not in ZL rats. Chronic ebselen treatment of ZDF rats restored renal tissue levels of glutathione and tetrahydrobiopterin; prevented significant accumulation of dihydrobiopterin, lipid peroxidation products, and 3-NT-modified proteins; and ameliorated focal and segmental sclerosis, proteinuria, and fall in creatinine clearance without affecting mean BP, body weight, and blood glucose, compared with the untreated ZDF rats. Chronic ebselen therapy also ameliorated vasculopathy with lipid deposits and tubulointerstitial scarring, inflammation, and upregulated alpha smooth muscle actin expression. These findings suggest that ZDF rats develop a progressive nephropathy with glomerular, vascular, and tubulointerstitial pathology. Oxidative and nitrosative stress predates the nephropathy, which is improved by peroxinitrite scavenger ebselen, and thus considered its cause and not consequence. PMID- 15339989 TI - Bradykinin decreases plasminogen activator inhibitor-1 expression and facilitates matrix degradation in the renal tubulointerstitium under angiotensin-converting enzyme blockade. AB - A number of experimental and clinical investigations support the notion that angiotensin-converting enzyme inhibitor (ACEi) and angiotensin II type 1 receptor blocker (ARB) compounds attenuate renal fibrosis. Fibrosis can be attenuated by either suppressing matrix formation or facilitating matrix degradation. In this study, drugs of ACEi and ARB classes were tested for their ability to facilitate matrix degradation in the kidney. A murine model system in which cyclosporin A (CsA) treatment for a specified period caused interstitial matrix deposition in the kidney was used. CsA was then discontinued, and experimental procedures were initiated to investigate matrix degradation. Benazepril, an ACEi, facilitated matrix degradation via the bradykinin (BK) B2 receptor on tubular epithelial cells in the kidney, whereas CGP-48933, an ARB, did not. In this murine model of CsA nephropathy under ACE blockade, plasminogen activator inhibitor-1 (PAI-1) expression was decreased in tubular epithelial cells, possibly leading to conversion of plasminogen to plasmin by plasminogen activator and subsequent activation of matrix metalloproteinases. These findings were confirmed in this study by measurements of plasmin activity, collagenolytic activity, and matrix metalloproteinase activities in the kidneys. In tubular epithelial cells stimulated in vitro, BK suppressed PAI-1 gene expression. All of these results suggest that ACEi can decrease PAI-1 expression via BK, thereby facilitating matrix degradation via activation of degradative enzymes to reduce interstitial matrix deposition. PMID- 15339990 TI - Complement activation regulates the capacity of proximal tubular epithelial cell to stimulate alloreactive T cell response. AB - Tissue expression of C3 is an unexpected regulator of the alloimmune response in mouse kidney transplantation. It is unclear, however, whether a direct or an indirect action of complement on the host immune response is involved. Also unknown is which of the complement effector products, cleaved C3, cleaved C5, or C5b-9, is responsible. Proximal tubular epithelial cells (PTEC) not only constitute a major target of the alloimmune response but also produce substantial amounts of C3. This study investigated the property of mouse PTEC to stimulate alloreactive T cells in a complement-dependent manner. The proliferative and cytokine responses of primed alloreactive T cells were measured after exposure to donor-specific PTEC that had been pretreated with normal mouse serum, heat inactivated mouse serum, or complement- deficient (C3, C5, or C6) mouse sera to differentially deposit complement components. PTEC were able to stimulate alloreactive T cells in an antigen-specific manner. Complement activation leading to the deposition of cleaved C3 on PTEC enhanced the alloreactive T cell response. This complement-mediated stimulation of the T cell response was dependent on C3 but not on C5 or C6. The primary influence of tissue-bound complement was on CD4(+) T cells. Moreover, the effect of complement on alloreactive T cells was B7 dependent, shown by inhibition studies with CTLA4-Ig. These results suggest that donor epithelium-bound C3 can upregulate the alloimmune response. It is postulated that surface-bound C3 interacts with complement receptors on alloreactive T cells or on antigen presenting cells to increase allo-immune stimulation. PMID- 15339991 TI - Mechanism of action of donor-specific transfusion in inducing tolerance: role of donor MHC molecules, donor co-stimulatory molecules, and indirect antigen presentation. AB - Donor-specific transfusion (DST) can synergize with T cell co-stimulatory blockade in inducing tolerance in several transplant models, but the mechanism of action of DST is poorly characterized. This study used genetically altered mice in an established model of cardiac transplantation to study the role of MHC and co-stimulatory molecule expression on DST cells in mediating the immunomodulatory effects of DST. In addition, to examine the role of indirect antigen presentation in the effect of DST, experiments used recipient mice that do not express MHC class II molecules on peripheral antigen-presenting cells, but do have functional CD4(+) T cells (II(-)4(+)). As previously reported, treatment with DST from wild type donors in combination with CD154 blockade induced tolerance in wild-type recipients of cardiac allografts. Tolerance in this model is also induced despite the absence of MHC class I and II, CD40, or B7 molecules on transfused cells. In contrast, eliminating the indirect pathway using II(-)4(+) recipients blocked the induction of long-term cardiac allograft survival by DST. These results indicate that the indirect antigen recognition pathway mediates the immunomodulatory effect of DST in inducing transplantation tolerance in vivo. PMID- 15339992 TI - RhoA activation mediates phosphatidylinositol 3-kinase-dependent proliferation of human vascular endothelial cells: an alloimmune mechanism of chronic allograft nephropathy. AB - HLA class I ligation on graft endothelial cells (EC) has been shown to promote graft arteriosclerosis and chronic allograft nephropathy. This study investigated transcriptional and functional changes mediated by anti-HLA antibodies (Ab), developed by transplant recipient, on vascular renal EC. For mimicking interactions that occur between alloantibodies and graft endothelium, HLA-typed primary cultures of human EC were incubated in vitro in the presence of monomorphic or polymorphic anti-HLA class I Ab. Gene expression analysis identified the upregulation of several molecules involved in cell signaling and proliferation, including the GTP-binding protein RhoA. It was demonstrated further that HLA class I ligation on EC induced a rapid translocation of RhoA to the cell membrane associated with F-actin stress fiber formation and cytoskeleton reorganization. Western blot analysis showed that anti-HLA class I Ab induced, in addition to RhoA, the activation of phosphatidylinositol 3-kinase, reflected by the phosphorylation of Akt (Ser473) and GSK3beta (Ser9), in EC. C3 exoenzyme, an inhibitor of RhoA, inhibited RhoA translocation in response to HLA class I ligation and reduced phosphatidylinositol 3-kinase activation. EC proliferation and cell cycle progression, examined by 5,6-carboxyfluorescein diacetate succinimidyl ester staining, demonstrated that anti-HLA-induced EC proliferation was efficiently prevented by the 3-hydroxy-3-methylglutaryl CoA reductase inhibitor simvastatin (0.1 micromol/L) through inhibition of RhoA geranylgeranylation. Taken together, these findings support the conclusion that RhoA is a key mediator of signaling pathways that lead to cytoskeletal reorganization and EC proliferation in response to alloantibodies that bind to HLA class I and demonstrate the specific and potent inhibitory effect of simvastatin on allostimulated EC growth. PMID- 15339993 TI - New aspects in the pathophysiology of preeclampsia. AB - Preeclampsia, the de novo occurrence of hypertension and proteinuria after the 20th week of gestation, continues to exert an inordinate toll on mothers and children alike. Recent clinical trials, new physiologic insights, and novel observations on pathogenesis have altered the thinking about preeclampsia. The mechanisms surrounding relaxin and its effects on the circulation and on matrix metalloproteinases have been elucidated. The growth factor's receptor, fms-like tyrosine kinase 1, has been shown to exist in a soluble form that is able to inactivate vascular endothelial-derived growth factor and human placental growth factor. Compelling evidence has been brought forth suggesting that fms-like tyrosine kinase 1 is a circulating factor that can cause preeclampsia. Preeclamptic women have high circulating levels of asymmetric dimethyl arginine that could account for the generalized endothelial dysfunction observed in preeclampsia. Preeclamptic women also produce novel autoantibodies that may serve to activate angiotensin receptors. These new observations raise the possibility that the treatment of preeclamptic women will soon be improved. PMID- 15339994 TI - Oxidative stress is increased in critically ill patients with acute renal failure. AB - Patients with acute renal failure (ARF) experience a high mortality rate. Dysregulated inflammation and altered metabolism may increase oxidative stress in ARF patients. Thirty-eight patients who met the Program to Improve Care in Acute Renal Disease (PICARD) Study inclusion criteria underwent plasma protein oxidation and plasma cytokine measurements. For comparison, similar measurements were also performed in 21 critically ill patients without ARF, 28 patients with ESRD, and 49 healthy subjects. Plasma protein thiol oxidation was measured by spectrophotometry. Plasma protein carbonyl content and cytokine concentrations were measured by ELISA. Plasma protein thiol oxidation and carbonyl content were markedly different in ARF patients compared with healthy subjects, ESRD patients, and critically ill patients (P < 0.001 in all cases). There were significant but less marked differences in plasma protein oxidation between ESRD patients and critically ill patients compared with healthy subjects. Plasma protein thiol oxidation in ARF patients improved with dialysis (P < 0.001); however, there was significant plasma oxidant reaccumulation during the interdialytic period (P < 0.001) not due to rebound equilibration of compartmentalized solutes. Plasma proinflammatory cytokine levels were significantly higher (P < 0.05) in ARF patients and critically ill patients than in healthy subjects. Plasma protein oxidation is markedly increased in ARF patients compared with healthy subjects, ESRD patients, and critically ill patients. Increased oxidative stress may be an important target for nutritional and pharmacologic therapy in ARF patients. PMID- 15339995 TI - Genomewide linkage analysis to serum creatinine, GFR, and creatinine clearance in a community-based population: the Framingham Heart Study. AB - Kidney disease is a risk factor for the development of cardiovascular disease, all-cause mortality, and ESRD. It is not known to what extent genetic factors play a role in the development of kidney disease in the general population. Multipoint variance components linkage analysis was performed using Genehunter on 330 families from the Framingham Heart Study offspring cohort, using a 10-cM genomewide scan for serum creatinine, GFR, and creatinine clearance (CRCL) measured from 1998 to 2001. GFR was estimated using the simplified Modification of Diet in Renal Disease Study equation, and CRCL was estimated using the Cockcroft-Gault equation. Covariates in the adjustment included age, gender, body mass index, diabetes, systolic BP, hypertension treatment, tobacco use, and HDL cholesterol. Overall, 1224 subjects (52% women), mean age 59, were available for analysis. Mean creatinine was 0.87 mg/dl, mean GFR was 87 ml/min per 1.73 m(2), and mean creatinine clearance was 100 ml/min. The multivariable-adjusted heritability estimates for creatinine, GFR, and CRCL were 0.29, 0.33, and 0.46, respectively. The peak log of the odds ratio (LOD) scores for serum creatinine, GFR, and CRCL were 2.28 at 176 cM on chromosome 4, 2.19 at 78 cM on chromosome 4, and 1.91 at 103 cM on chromosome 3, respectively. In a community-based sample, measures of serum creatinine, GFR, and CRCL are heritable, suggesting an underlying genetic component. These results also provide suggestive evidence for linkage to measures of kidney function. Further research is necessary to identify the genes involved in the development of kidney disease and to understand their roles in this complex process. PMID- 15339996 TI - "Renalism": inappropriately low rates of coronary angiography in elderly individuals with renal insufficiency. AB - Higher risk patients (including the elderly) receive more conservative therapy for cardiovascular diseases, even though the relative benefits of therapy tend to be greater. The perceived risk of radiocontrast-associated nephrotoxicity may influence the provision of coronary angiography and subsequent revascularization, especially among individuals with chronic kidney disease (CKD). The aim of this study was to determine whether there is excessive variation in the provision of coronary angiography after acute myocardial infarction on the basis of the presence of CKD and whether there is an association between angiography and mortality. Elderly (age 65 to 89 yr) individuals with acute myocardial infarction from the Cooperative Cardiovascular Project were classified by the presence or absence of CKD (defined as a baseline serum creatinine of 1.5 to 5.0 mg/dl). In CKD patients, the propensity to undergo coronary angiography was determined and the effect of coronary angiography on mortality was estimated using multivariable logistic regression and stratification. Mortality was significantly higher with CKD (52.6 versus 26.4%). Fewer patients with CKD underwent coronary angiography (25.2 versus 46.8%) despite the observation that a similar proportion of patients were deemed appropriate for angiography by standard, published criteria. When limiting the analysis to CKD patients who are considered appropriate, the multivariable estimate of the odds of death associated with coronary angiography was 0.58 (95% confidence interval, 0.50 to 0.67). With adjustment using propensity scores, the odds ratio averaged across propensity score quintiles was 0.62 (95% confidence interval, 0.54 to 0.70). Results were qualitatively similar when patients were stratified by CKD stage IV (estimated GFR <30 ml/min per 1.73 m(2)). There is a large relative decrease in utilization of coronary angiography among patients with CKD. Alteration in practice because of an aversion to the risk of radiocontrast-associated nephrotoxicity ("renalism") is inappropriate, even if the true relative benefit of invasive strategies is a fraction of what is estimated here. PMID- 15339997 TI - Retinal microvascular abnormalities and renal dysfunction: the atherosclerosis risk in communities study. AB - Microvascular disease has been linked with renal dysfunction in patients with diabetes. The aim of this study was to examine the association of retinal microvascular abnormalities to renal dysfunction among participants of the Atherosclerosis Risk in Communities Study, a population-based investigation in four U.S. communities. At the third examination (1993 to 1995), retinal photography was performed and the presence of retinal microvascular abnormalities was documented using a standard grading protocol. Renal dysfunction was defined as an increase in serum creatinine of at least 0.4 mg/dl or a death or hospitalization as a result of chronic kidney disease between the second (1990 to 1992) and fourth (1996 to 1998) examinations. Among 10,056 people who were included in the study, 270 (2.7%) developed renal dysfunction. After controlling for age, gender, race, diabetes, BP, and other risk factors, individuals with retinopathy (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.4 to 2.8), microaneurysms (OR, 2.0; 95% CI, 1.3 to 3.1), retinal hemorrhages (OR, 2.6; 95% CI, 1.6 to 4.0), soft exudates (OR, 2.7; 95% CI, 1.6 to 4.8), and arteriovenous nicking (OR, 1.4; 95% CI, 1.0 to 1.9) were more likely to develop renal dysfunction than individuals without these abnormalities. Retinal microvascular abnormalities are associated with renal dysfunction, suggesting that common systemic microvascular processes may underlie the development of microvascular damage in the eye and kidneys. PMID- 15339998 TI - Is there an epidemic of HIV Infection in the US ESRD program? AB - Surveys revealed increases in the prevalence of HIV-infected patients in the US end-stage renal disease (ESRD) program in the 1980s and early 1990s, with clustering in young black men 25 to 44 yr old. Since the availability of highly active antiretroviral therapy in 1996, the prognosis of HIV-infected patients has improved, and therapy has been shown to change the course of classic HIV associated nephropathy. We used the United States Renal Data System database to determine if the incidence and prevalence of HIV-infected patients with renal disease has increased in the ESRD program, by means of principal diagnoses and comorbid AIDS-defining diagnoses. As the number of US patients living with AIDS increased 57% from 214,711 in 1995 to 337,017 in 2000, and the number of incident ESRD patients increased 29.9% from 72,827 to 94,602, the number of incident HIV infected patients increased only by 3.5%, from 1133 to 1171. Over this time, the percentage of incident ESRD patients with HIV infection fell from 1.56% to 1.24%. Among black men 25 to 44 yr of age, HIV infection as a proportion of incident ESRD cases fell from 8.5% to 6.2% from 1995 to 2000. The incident rate per million of AIDS or HIV infection in black men aged 25 to 44 fell from 107 in 1995 to 78 per million in 2000. The incidence rate for HIV-infected women in the ESRD program rose 14% while it declined 7% in men. Almost 2000 HIV-infected women, or 28.8% of the population, have initiated therapy for ESRD with hemodialysis. The number of prevalent cases increased in absolute numbers 81.3% from 2687 to 4871 (0.90% to 1.16% of the ESRD program). One-year survival rates for HIV-infected incident ESRD patients increased from 53.1% to 67.1% from 1995 to 2000. Although these values may be underestimates because of underreporting due to confidentiality concerns and lack of biopsy confirmation, we conclude that although the prevalence of HIV infection is increasing in the US ESRD population, the increase as a proportion of the program is minimal and is due to better survival after development of renal failure. The incidence of HIV infection in the US ESRD program is stable. Highly active antiretroviral therapy may be responsible for the change in epidemiology of HIV infection in the US ESRD program. PMID- 15339999 TI - Effect of intravenous ascorbic acid medication on serum levels of soluble transferrin receptor in hemodialysis patients. AB - Intravenous ascorbic acid (IVAA) medication has been shown to facilitate iron release from inert depots and subsequently circumvent the defective iron utilization in chronic hemodialysis (HD) patients who are treated with recombinant human erythropoietin (rHuEPO). This study focuses on the effects of IVAA supplementation on serum concentrations of soluble transferrin receptors (TfR) on the basis of the hypothesis that an increase of labile iron in the cytosol will lead to inhibition of TfR expression. First, 138 HD patients were studied to evaluate the interrelation between serum TfR and iron status. In a stepwise multivariate analysis, serum EPO and transferrin saturation (TSAT) were the two independent predictors for serum TfR in HD patients (r(2) = 0.510, P < 0.001). Further analyses showed that the lower the serum EPO and the higher the TSAT, the lower the serum TfR in HD patients who are on maintenance rHuEPO treatment. Second, 36 HD patients were recruited in a randomized, controlled study to receive IVAA (total dose of 2000 mg) or normal saline (placebo) medication. Serum levels of TfR, EPO, and ferritin and TSAT were measured at baseline and within 7 d after starting IVAA or placebo. There were no significant changes in serum EPO and ferritin levels in patients who received either IVAA (n = 18) or placebo (n = 18). Serum TfR levels (P < 0.001) significantly declined with a parallel rise in TSAT (P < 0.05) as compared with presupplemental values within 7 d in IVAA patients before any apparent alteration in hematocrit values, but the changes were not observed in the placebo group. The trend of decreased serum TfR and increased TSAT was similar in IVAA patients with ferritin of <500 microg/L or >500 microg/L. It is concluded that ascorbic acid status can significantly decrease serum TfR concentrations and increase percentage of TSAT, probably through alterations in intracellular iron metabolism. PMID- 15340000 TI - Presence of a failed kidney transplant in patients who are on hemodialysis is associated with chronic inflammatory state and erythropoietin resistance. AB - Patients returning to hemodialysis (HD) after failure of their kidney transplant suffer from high morbidity and mortality rates. It is common practice to keep failed kidney transplants in place. It is not known if these failed kidney transplants induce an inflammatory state that contributes to morbidity and mortality. In a single facility, patients starting on HD with failed kidney transplant were identified (Group A) and screened for the presence of chronic inflammatory state. Those with clinical symptoms attributed to the failed allograft (Group A1) were not offered transplant nephrectomy unless deemed necessary during follow-up. Their clinical and laboratory data were followed up for 6 months. Similar data were obtained from a group of incident HD patients (Group B). Forty-three patients had a failed Kidney transplant (Group A). Of these, 29 comprised Group A1 and 14 Group A2. Group B comprised 121 patients. In comparison with Group B, Group A exhibited worse anemia and erythropoietin resistance index (ERI), had lower serum albumin and prealbumin, and higher CRP. Group A1 had lower Hb and higher ferritin, CRP, and ESR in comparison with Group A2. Following transplant nephrectomy, Group A1 had improvement in ERI, serum albumin, prealbumin, ferritin, fibrinogen, CRP, and ESR. At 6 months, Group A1 had higher Hb and serum albumin levels, and lower CRP and ERI in comparison with Group A2. Group B parameters showed no change during follow-up. Patients returning to HD following failure of their kidney transplant suffer from a chronic inflammatory state. Resection of failed transplants in symptomatic patients is associated with amelioration of markers of chronic inflammation. Transplant nephrectomy should be considered a treatment option for patients with failed kidney transplants, especially if they exhibit signs and symptoms of chronic inflammatory state. PMID- 15340001 TI - Statin induced proteinuria: renal injury or renoprotection? PMID- 15340002 TI - Bradykinin and renal fibrosis: have we ACE'd it? PMID- 15340003 TI - Repeated amphetamine couples norepinephrine transporter and calcium channel activities in PC12 cells. AB - Repeated intermittent amphetamine enhances efflux of dopamine through the dopamine transporter in rat basal ganglia and through the norepinephrine transporter in rat pheochromocytoma PC12 cells. Extracellular Ca2+ is required for the detection of this enhancement in the rat. In this study, we examined the role of Ca2+ and Ca2+ channels in the enhanced amphetamine-induced dopamine efflux that develops in PC12 cells following repeated intermittent amphetamine. Repeated pretreatment of PC12 cells with 1 microM amphetamine followed by a drug free period increased amphetamine-induced efflux of dopamine compared with controls. The enhancement in amphetamine-induced dopamine efflux depended upon the presence of extracellular Ca2+ and was inhibited by the blockade of N-type and L-type Ca2+ channels. The enhanced dopamine efflux was not altered by tetanus toxin or reserpine, treatments that abrogate synaptic vesicle-mediated, exocytotic dopamine efflux. Measurement of intracellular Ca2+ concentrations using fura-2/acetoxymethyl ester revealed that amphetamine increased intracellular Ca2+ by a transporter-dependent mechanism. In amphetamine pretreated cells, amphetamine elicited a greater increase in intracellular Ca2+; this increase depended upon the presence of extracellular Ca2+ and N- and L-type Ca2+ channel activity. The enhanced amphetamine-induced dopamine efflux requires Ca2+/calmodulin kinase activity. In vehicle-treated cells, 1 microM amphetamine inhibited the calmodulin kinase activity although it did not in amphetamine pretreated cells. This study suggests that repeated intermittent amphetamine couples norepinephrine transporter activity and Ca2+ signaling. PMID- 15340004 TI - Increased expression of the sodium transporter BSC-1 in spontaneously hypertensive rats. AB - The purpose of this study was to compare the expression of BSC-1 (bumetanide sensitive Na+-K+-2Cl- cotransporter) in kidneys of spontaneously hypertensive rats (SHR) versus Wistar-Kyoto (WKY) rats by immunoblotting and reverse transcription-polymerase chain reaction. To determine the specificity of any observed changes in BSC-1 expression, we also compared expression of the thiazide sensitive Na+-Cl- cotransporter (TSC), the type-3 Na+-H+ exchanger (NHE-3), Na+ K+-ATPase-alpha1, the inwardly rectifying K+ channel (ROMK-1), the type-1 Na+ HCO3- cotransporter (NBC-1), aquaporin-1, and aquaporin-2. Analyses were performed on outer cortex, outer medulla, and inner medulla. BSC-1 protein was detected in outer medulla and was markedly (6-fold) higher in SHR. TSC protein was detected in the cortex and was not overexpressed in SHR. Aquaporin-1 protein was detected in all three regions and was not overexpressed in SHR. Aquaporin-2 and ROMK-1 proteins were detected in all three regions, but were moderately elevated (2-fold) only in the SHR inner medulla. Na+-K+-ATPase and NHE-3 proteins were detected in all three regions. Na+-K+-ATPase-alpha1 was modestly (25%) increased in SHR outer and inner medulla, whereas NHE-3 was moderately (2-fold) increased in the SHR cortex and inner medulla. NBC-1 protein was detected only in the cortex and was higher (2-fold) in SHR. mRNA levels of BSC-1, aquaporin-2, and ROMK-1 were not elevated in SHR, indicating a post-translational mechanism of protein overexpression. High-dose furosemide increased fractional sodium excretion more in SHR than WKY (3-fold). We conclude that increased expression of BSC-1, and to a lesser extent, aquaporin-2, ROMK-1, NHE-3, and NBC-1 may contribute to the pathogenesis of hypertension in the SHR. PMID- 15340005 TI - Assessment of the relative reinforcing strength of cocaine in socially housed monkeys using a choice procedure. AB - Position in the social hierarchy can influence brain dopamine function and cocaine reinforcement in nonhuman primates during early cocaine exposure. With prolonged exposure, however, initial differences in rates of cocaine self administration between dominant and subordinate monkeys dissipate. The present studies used a choice procedure to assess the relative reinforcing strength of cocaine in group-housed male cynomolgus monkeys with extensive cocaine self administration histories. Responding was maintained under a concurrent fixed ratio 50 schedule of food and cocaine (0.003-0.1 mg/kg per injection) presentation. Responding on the cocaine-associated lever increased as a function of cocaine dose in all monkeys. Although response distribution was similar across social rank when saline or relatively low or high cocaine doses were the alternative to food, planned t tests indicated that cocaine choice was significantly greater in subordinate monkeys when choice was between an intermediate dose (0.01 mg/kg) and food. When a between-session progressive-ratio procedure was used to increase response requirements for the preferred reinforcer (either cocaine or food), choice of that reinforcer decreased in all monkeys. The average response requirement that produced a shift in response allocation from the cocaine-associated lever to the food-associated lever was higher in subordinates across cocaine doses, an effect that trended toward significance (p = 0.053). These data indicate that despite an extensive history of cocaine self administration, most subordinate monkeys were more sensitive to the relative reinforcing strength of cocaine than dominant monkeys. PMID- 15340006 TI - Lack of specific amyloid-beta(1-42) suppression by nonsteroidal anti-inflammatory drugs in young, plaque-free Tg2576 mice and in guinea pig neuronal cultures. AB - Recent studies indicating that some nonsteroidal anti-inflammatory drugs (NSAIDs) selectively modulate gamma-secretase cleavage of amyloid precursor protein (APP) while sparing Notch processing have generated interest in discovery of novel gamma-secretase modulators with the "NSAID-like" efficacy profile. The objective of the present studies was to compare the efficacy of a subset of NSAIDs with previously reported classical gamma-secretase inhibitors LY-411575 [N(2)-[(2S)-2 (3,5-difluorophenyl)-2-hydroxyethanoyl]-N(1)-[(7S)-5-methyl-6-oxo-6,7-dihydro-5H dibenzo[b,d]azepin-7-yl]-L-alaninamide]and DAPT [N-[N- (3,5-difluorophenacetyl)-L alanyl]-S-phenylglycine t-butyl ester] in Tg2576 mice. Flurbiprofen (10 and 25 mg/kg/day) was overtly toxic and elicited significant (but nonselective) reductions in both Abeta(1-40) and Abeta(1-42) in the plasma in one of two studies. Flurbiprofen also produced a small reduction in Abeta(1-40) in the cortex at 25 mg/kg/day but did not affect Abeta levels in hippocampus or cerebrospinal fluid. Ibuprofen and sulindac sulfide were neither overtly toxic nor efficacious at doses up to 50 mg/kg/day. The effects of NSAIDs LY-411575 and DAPT were tested in guinea pig embryonic neuronal cultures to determine whether the selective reductions in Abeta(1-42) observed in cell lines overexpressing human mutant APP can be reproduced in a neuronal model of physiological Abeta production and secretion. Flurbiprofen and sulindac nonselectively reduced Abeta(1-40) and Abeta(1-42) at concentrations > or =125 microM, although cytotoxicity was noted at > or =250 microM sulindac. Ibuprofen had no effect at concentrations up to 500 microM. In contrast, DAPT and LY-411575 potently and completely inhibited Abeta(1-40), Abeta(1-42), and Abeta(1-38) in the absence of cytotoxicity. The divergence of the present data from published reports raises the need to examine the conditions necessary to perceive selective Abeta(1-42) reduction by NSAIDs in neuronal tissue. PMID- 15340007 TI - Outer dense fibre protein 2 (ODF2) is a self-interacting centrosomal protein with affinity for microtubules. AB - Outer dense fibre protein 2 (ODF2) is a major protein of sperm tail outer dense fibres which are prominent sperm tail-specific cytoskeletal structures. Moreover, ODF2 was also identified as a widespread component of the centrosomal scaffold and was found to associate preferentially with the appendages of the mother centriole [Nakagawa, Y., Yamane, Y., Okanoue, T., Tsukita, S. and Tsukita, S. (2001) Mol. Biol. Cell 12, 1687-1697]. Secondary structure predictions indicated ODF2 as an overall coiled-coil protein with a putative fibre forming capacity. To investigate its potential functions in generating the centrosomal scaffold and in microtubule nucleation we asked whether ODF2 is able to form a fibrillar structure by self-association in vivo and if it interacts with microtubules. By cytological investigation of transfected mammalian cells expressing ODF2-GFP fusion proteins and in vitro coprecipitation assays we could demonstrate that ODF2 is a self-interacting protein that forms a fibrillar structure partially linked to the microtubule network. Microtubule cosedimentation and coprecipitation assays indicated ODF2 as a microtubule-associated protein. However, we could not demonstrate a direct interaction of ODF2 with tubulin, suggesting that binding of endogenous ODF2 to the axonemal as well as to centrosomal microtubules may be mediated by, as yet, unknown proteins. PMID- 15340008 TI - Requirement for Schizosaccharomyces pombe Top3 in the maintenance of chromosome integrity. AB - In Schizosaccharomyces pombe, topoisomerase III is encoded by a single gene, top3(+), which is essential for cell viability and proper chromosome segregation. Deletion of rqh1(+), which encodes the sole RecQ family helicase in S. pombe, suppresses the lethality caused by loss of top3. Here, we provide evidence suggesting that the lethality in top3 mutants is due to accumulation of aberrant DNA structures that arise during S phase, as judged by pulsed-field gel electrophoresis. Using a top3 shut-off strain, we show here that depletion of Top3 activates the DNA damage checkpoint associated with phosphorylation of the checkpoint kinase Chk1. Despite activation of this checkpoint, top3 cells exit the arrest but fail to undergo faithful chromosome segregation. However, these mitotic defects are secondary to chromosomal abnormalities that lead to the lethality, because advance into mitosis did not adversely affect cell survival. Furthermore, top3 function is required for maintenance of nucleolar structure, possibly due to its ability to prevent recombination at the rDNA loci. Our data are consistent with the notion that Top3 has a key function in homologous recombinational repair during S phase that is essential for ensuring subsequent fidelity of chromosome segregation. PMID- 15340009 TI - The role of myosin heavy chain phosphorylation in Dictyostelium motility, chemotaxis and F-actin localization. AB - To assess the role of myosin II heavy chain (MHC) phosphorylation in basic motility and natural chemotaxis, the Dictyostelium mhcA null mutant mhcA(-), mhcA(-) cells rescued with a myosin II gene that mimics the constitutively unphosphorylated state (3XALA) and mhcA(-) cells rescued with a myosin II gene that mimics the constitutively phosphorylated state (3XASP), were analyzed in buffer and in response to the individual spatial, temporal and concentration components of a cAMP wave using computer-assisted methods. Each mutant strain exhibited unique defects in cell motility and chemotaxis. Although mhcA(-) cells could crawl with some polarity and showed chemotaxis with highly reduced efficiency in a spatial gradient of cAMP, they were very slow, far less polar and more three-dimensional than control cells. They were also incapable of responding to temporal gradients of cAMP, of chemotaxis in a natural wave of cAMP or streaming late in aggregation. 3XASP cells were faster and chemotactically more efficient than mhcA(-) cells, but still incapable of responding to temporal gradients of cAMP, chemotaxis in natural waves of cAMP or streaming late in aggregation. 3XALA cells were fast, were able to respond to temporal gradients of cAMP, and responded to natural waves of cAMP. However, they exhibited a 50% reduction in chemotactic efficiency, could not stream late in aggregation and could not enter the streams of control cells in mixed cultures. F-actin staining further revealed that while the presence of unphosphorylated MHC was essential for the increase in F-actin in the cytoplasm in response to the increasing temporal gradient of cAMP in the front of a natural wave, the actual dephosphorylation event was essential for the associated increase in cortical F actin. The results of these studies indicate that MHC phosphorylation dephosphorylation, like myosin II regulatory light chain phosphorylation dephosphorylation, represents a potential downstream target of the regulatory cascades emanating from the different phases of the wave. PMID- 15340010 TI - v-Src accelerates spontaneous motility via phosphoinositide 3-kinase, phospholipase C and phospholipase D, but abrogates chemotaxis in Rat-1 and MDCK cells. AB - In Rat-1 fibroblasts, v-Src causes a profound remodelling of cortical actin cytoskeleton. This transformation includes membrane ruffling, a hallmark of the leading edge in migrating cells, and results from activation of phosphoinositide 3-kinase (PI 3-kinase), phospholipase C (PLC) and phospholipase D (PLD). We therefore reexamined whether motility is constitutively triggered by v-Src and studied whether this response is controlled by the same signalling pathway. The study was performed using Rat-1/tsLA29 and MDCK/tsLA31 cells, each harbouring a different thermosensitive v-Src kinase, active at 34 degrees C but inactivated at 40 degrees C. In both cell lines, overnight v-Src activation induced transformation and accelerated spontaneous motility by approximately twofold, as evidenced by wound-healing assay and by single-cell track, time-lapse recording in Dunn chambers. Inhibitors of PI 3-kinase, PLC and PLD selectively abrogated acceleration of motility by v-Src. Since mechanisms that co-ordinate spontaneous, as distinct from oriented, cell migration are separable, we further analysed in Dunn chambers chemotactic response of Rat-1/tsLA29 cells to PDGF and of MDCK/tsLA31 cells to EGF. In both cases, v-Src decreased the steady-state level of growth factor receptors at the cell surface twofold, and abrogated movement directionality at comparable level of occupancy as in non-transformed cells. The burst of pinocytosis in response to growth factors was also abolished by v-Src. Altogether, these results indicate that v-Src triggers motility in a PI 3-kinase , PLC- and PLD-dependent manner, but abrogates directionality by suppressing polarised signalling downstream of growth factor receptors. PMID- 15340011 TI - Arabidopsis VARIEGATED 3 encodes a chloroplast-targeted, zinc-finger protein required for chloroplast and palisade cell development. AB - The stable, recessive Arabidopsis variegated 3 (var3) mutant exhibits a variegated phenotype due to somatic areas lacking or containing developmentally retarded chloroplasts and greatly reduced numbers of palisade cells. The VAR3 gene, isolated by transposon tagging, encodes the 85.9 kDa VAR3 protein containing novel repeats and zinc fingers described as protein interaction domains. VAR3 interacts specifically in yeast and in vitro with NCED4, a putative polyene chain or carotenoid dioxygenase, and both VAR3 and NCED4 accumulate in the chloroplast stroma. Metabolic profiling demonstrates that pigment profiles are qualitatively similar in wild type and var3, although var3 accumulates lower levels of chlorophylls and carotenoids. These results indicate that VAR3 is a part of a protein complex required for normal chloroplast and palisade cell development. PMID- 15340012 TI - Signaling pathways controlling primordial germ cell migration in zebrafish. AB - During their migration, zebrafish primordial germ cells (PGCs) rely on directional cues provided by the chemokine SDF-1a, whose receptor is CXCR4b. The molecular mechanisms whereby CXCR4b activation is interpreted intracellularly into directional migration are not known. Here we investigate the role of two important biochemical pathways -- G-protein-dependent and phosphoinositide 3 kinase (PI3K)-dependent signaling -- in directing PGC migration in zebrafish. We show that G proteins of the Gi family are essential for directional migration but not for PGC motility. Inhibition of PI3K signaling in PGCs slows down their migration and leads to abnormal cell morphology as well as to reduced stability of filopodia. Invariably, during directed PGC migration, the distribution of the products of PI3K activity - phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P(3)] and/or phosphatidylinositol (3,4)bisphosphate [PtdIns(3,4)P(2)] -- is not polarized, and reducing the level of these 3 phosphoinositides does not affect the ability of PGCs to migrate directionally. We therefore conclude that Gi-dependent signaling is essential for directional migration, whereas the PI3K pathway is important for the actual motility of PGCs. PMID- 15340013 TI - The RacGEF Tiam1 inhibits migration and invasion of metastatic melanoma via a novel adhesive mechanism. AB - Rho-like GTPases such as RhoA, Rac1 and Cdc42 are key regulators of actin dependent cell functions including cell morphology, adhesion and migration. Tiam1 (T lymphoma invasion and metastasis 1), a guanine nucleotide exchange factor that activates Rac, is an important regulator of cell shape and invasiveness in epithelial cells and fibroblasts. Overexpression of Tiam1 in metastatic melanoma cells converted the constitutive mesenchymal phenotype into an epithelial-like phenotype. This included the induction of stringent cell-cell contacts mediated by the Ig-like receptor ALCAM (activated leukocyte cell adhesion molecule) and actin redistribution to cell-cell junctions. This phenotypic switch was dependent on increased Rac but not Rho activity, and on the redistribution and adhesive function of ALCAM, whereas cadherins were not involved. Although cell proliferation was significantly enhanced, the gain of cell-cell junctions strongly counteracted cell motility and invasion as shown for two- and three dimensional collagen assays as well as invasion into human skin reconstructs. The reverse transition from mesenchymal invasive to a resident epithelial-like phenotype implicates a role for Tiam1/Rac signaling in the control of cell-cell contacts through a novel ALCAM-mediated mechanism. PMID- 15340014 TI - Role for Rab7 in maturation of late autophagic vacuoles. AB - The small GTP binding protein Rab7 has a role in the late endocytic pathway and lysosome biogenesis. The role of mammalian Rab7 in autophagy is, however, unknown. We have addressed this by inhibiting Rab7 function with RNA interference and overexpression of dominant negative Rab7. We show here that Rab7 was needed for the formation of preferably perinuclear, large aggregates, where the autophagosome marker LC3 colocalised with Rab7 and late endosomal and lysosomal markers. By electron microscopy we showed that these large aggregates corresponded to autophagic vacuoles surrounding late endosomal or lysosomal vesicles. Our experiments with quantitative electron microscopy showed that Rab7 was not needed for the initial maturation of early autophagosomes to late autophagic vacuoles, but that it participated in the final maturation of late autophagic vacuoles. Finally, we showed that the recruitment of Rab7 to autophagic vacuoles was retarded in cells deficient in the lysosomal membrane proteins Lamp1 and Lamp2, which we have recently shown to accumulate late autophagic vacuoles during starvation. In conclusion, our results showed a role for Rab7 in the final maturation of late autophagic vacuoles. PMID- 15340015 TI - Light-dependent subcellular translocation of Gqalpha in Drosophila photoreceptors is facilitated by the photoreceptor-specific myosin III NINAC. AB - We examine the light-dependent subcellular translocation of the visual G(q)alpha protein between the signaling compartment, the rhabdomere and the cell body in Drosophila photoreceptors. We characterize the translocation of G(q)alpha and provide the first evidence implicating the involvement of the photoreceptor specific myosin III NINAC in G(q)alpha transport. Translocation of G(q)alpha from the rhabdomere to the cell body is rapid, taking less than 5 minutes. Higher light intensities increased the quantity of G(q)alpha translocated out of the rhabdomeres from 20% to 75%, consistent with a mechanism for light adaptation. We demonstrate that translocation of G(q)alpha requires rhodopsin, but none of the known downstream phototransduction components, suggesting that the signaling pathway triggering translocation occurs upstream of G(q)alpha. Finally, we show that ninaC mutants display a significantly reduced rate of G(q)alpha transport from the cell body to the rhabdomere, suggesting that NINAC might function as a light-dependent plus-end motor involved in the transport of G(q)alpha. PMID- 15340016 TI - Pentamidine-induced long QT syndrome and block of hERG trafficking. AB - The diamidine pentamidine is used to treat leishmaniasis, trypanosomiasis, and Pneumocystis carinii pneumonia. Treatment may be accompanied by prolongation of the QT interval of the electrocardiogram and torsades de pointes tachycardias. Up to now, it has been thought that therapeutic compounds causing QT prolongation are associated with direct block of the cardiac potassium channel human ether a go-go-related gene (hERG), which encodes the alpha subunit of cardiac I(Kr) currents. We show that pentamidine has no acute effects on currents produced by hERG, KvLQT1/mink, Kv4.3, or SCNA5. Cardiac calcium currents and the guinea pig cardiac action potential were also not affected. After overnight exposure, however, pentamidine reduced hERG currents and inhibited trafficking and maturation of hERG with IC(50) values of 5 to 8 microM similar to therapeutic concentrations. Surface expression determined in a chemiluminescence assay was reduced on exposure to 10, 30, and 100 microM pentamidine by about 30, 40, and 70%, respectively. These effects were specific for hERG since expression of hKv1.5, KvLQT1/minK, and Kv4.3 was not altered. In isolated guinea pig ventricular myocytes, 10 microM pentamidine prolonged action potential duration APD(90) from 374.3 +/- 57.1 to 893.9 +/- 86.2 ms on overnight incubation. I(Kr) tail current density was reduced from 0.61 +/- 0.09 to 0.39 +/- 0.04 pA/pF. We conclude that pentamidine prolongs the cardiac action potential by block of hERG trafficking and reduction of the number of functional hERG channels at the cell surface. We propose that pentamidine, like arsenic trioxide, produces QT prolongation and torsades de pointes in patients by inhibition of hERG trafficking. PMID- 15340017 TI - Liver-targeted drug delivery using HepDirect prodrugs. AB - Targeting drugs to specific organs, tissues, or cells is an attractive strategy for enhancing drug efficacy and reducing side effects. Drug carriers such as antibodies, natural and manmade polymers, and labeled liposomes are capable of targeting drugs to blood vessels of individual tissues but often fail to deliver drugs to extravascular sites. An alternative strategy is to use low molecular weight prodrugs that distribute throughout the body but cleave intracellularly to the active drug by an organ-specific enzyme. Here we show that a series of phosphate and phosphonate prodrugs, called HepDirect prodrugs, results in liver targeted drug delivery following a cytochrome P450-catalyzed oxidative cleavage reaction inside hepatocytes. Liver targeting was demonstrated in rodents for MB06866 [(2R,4S)-9-[2-[4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2 yl]methoxyethyl]adenine (remofovir)], a Hep-Direct prodrug of the nucleotide analog adefovir (PMEA), and MB07133 [(2R,4S)-4-amino-1-[5-O-[2-oxo-4-(4-pyridyl) 1,3,2-dioxaphosphorinan-2-yl]-beta-d-arabinofuranosyl]-2(1H)-pyrimidinone], a HepDirect prodrug of cytarabine (araC) 5'-monophosphate. Liver targeting led to higher levels of the biologically active form of PMEA and araC in the liver and to lower levels in the most toxicologically sensitive organs. Liver targeting also confined production of the prodrug byproduct, an aryl vinyl ketone, to hepatocytes. Glutathione within the hepatocytes rapidly reacted with the byproduct to form a glutathione conjugate. No byproduct-related toxicity was observed in hepatocytes or animals treated with HepDirect prodrugs. A 5-day safety study in mice demonstrated the toxicological benefits of liver targeting. These findings suggest that HepDirect prodrugs represent a potential strategy for targeting drugs to the liver and achieving more effective therapies against chronic liver diseases such as hepatitis B, hepatitis C, and hepatocellular carcinoma. PMID- 15340018 TI - Home or hospital? Choices at the end of life. PMID- 15340019 TI - Mental capacity, legal competence and consent to treatment. AB - Deciding whether someone is legally competent to make decisions regarding their own treatment requires an assessment of their mental capacity. The assessed capacity required for legal competence increases with the seriousness of what is at stake. The usual explanation is that patient autonomy is being balanced against best interests. An alternative explanation, that we require greater room for error when the consequences are serious, implies a change to clinical practice and in the evidence doctors offer in court. PMID- 15340020 TI - Diagnosis of early rheumatoid arthritis: what the non-specialist needs to know. PMID- 15340021 TI - Glutamine supplements in the critically ill. PMID- 15340022 TI - Community hospitals: an under-recognized resource for palliative care. AB - In the UK there are concerns that, in certain groups of dying patients such as the old, those with non-cancer diagnoses and those in rural areas, the quality of care is unacceptably variable. There has been no systematic survey of the extent to which community hospitals provide general palliative care for such patients. Therefore, by means of a structured questionnaire we asked senior nurses/managers at all 478 community hospitals in the UK for information on staff expertise, facilities and specialist equipment, liaison arrangements with specialist palliative care providers, priorities, practice and policy in end-of-life care. Of the 346 hospitals (72%) that responded, only 28 were in urban areas. 73% of hospitals employed at least one nurse with additional training in palliative care, 72% had access to 24-hour specialist palliative care advice and 51% had separate overnight accommodation for relatives, but only 22% had designated palliative care beds. Most hospitals did not have written policies or guidelines for patient assessment or symptom control. These findings add to evidence that community hospitals represent an important resource to improve access to palliative care for groups that are currently under-served. PMID- 15340023 TI - Assessment of patient pain at colonoscopy: are nurses better than endoscopists? AB - The pain that patients recollect having experienced at colonoscopy is likely to influence uptake of the procedure. We used visual analogue scales to assess recollected pain shortly before discharge, and compared these scores with assessments by the endoscopist and the attending nurse. Data were complete for 426 procedures (90%). The mean perceived pain score for patients was 3.2, for endoscopists 2.8 and for nurses 3.1. On multivariate analysis, the endoscopists' assessments of pain had little predictive value over and above those of nurses, whereas nurses' assessments remained significant when adjusted for endoscopists' assessments. Nurses were more accurate than endoscopists in gauging the pain of colonoscopy. This may be because endoscopists are focused on the video monitor while nurses are focused on the patient. More active use of nurses' assessments might help keep pain to a minimum. PMID- 15340024 TI - Meningococcal chest pain. PMID- 15340025 TI - Bilateral spontaneous pneumothorax in a cannabis smoker. PMID- 15340026 TI - Optic neuropathy and orbital inflammatory mass after wasp stings. PMID- 15340027 TI - 'Football' contact lenses--an own goal. PMID- 15340028 TI - Vasculitic appendicitis. PMID- 15340029 TI - Uveitis in a patient receiving rifabutin for Crohn's disease. PMID- 15340030 TI - Prosthetic joint infection by cat scratch. PMID- 15340031 TI - Severe hypothyroidism after thalidomide treatment. PMID- 15340032 TI - Critical limb ischaemia associated with chemotherapy for breast cancer. PMID- 15340033 TI - 'Proper to the duty of a chirurgeon': Ambroise Pare and sixteenth century paediatric surgery. PMID- 15340034 TI - Lack of confidentiality with the Picture Archiving and Communication System (PACS). PMID- 15340035 TI - Specialist medical abbreviations as a foreign language. PMID- 15340037 TI - Death by footwear. PMID- 15340038 TI - Tuberculosis sanatoria regimens in the 1940s. PMID- 15340039 TI - Basic training in ophthalmology. PMID- 15340040 TI - The well-managed waiting list. PMID- 15340041 TI - Doxorubicin inhibits DNMT1, resulting in conditional apoptosis. AB - Chemotherapy using DNA intercalators is one of the most successful approaches to cancer treatment. Although DNA intercalators are believed to inhibit DNA polymerases and topoisomerases, resulting in the induction of apoptosis in tumor cells, other factors potentially inhibited by the anthracycline antibiotics remain to be elucidated. Herein, we show that the enzymatic activity of DNMT1, the primary DNA methyltransferase in mammalian cells, is inhibited by DNA intercalators, such as doxorubicin, in an in vitro assay. Enzymatic analyses indicate that doxorubicin inhibits the catalytic activity of DNMT1 via DNA intercalation. We also found that apoptosis was induced in DNMT1(+/+) HCT116 cells by only a limited range of doxorubicin dose, meaning that apoptotic cell death is "conditional" with respect to the concentration of the DNA intercalating drug. It is noteworthy that conditional apoptosis is not observed in human colorectal cancer cells lacking DNMT1 but can be induced in DNMT1(-/-) cells by transfection of a plasmid expressing DNMT1. Our results suggest that DNMT1 is one of the major targets of doxorubicin resulting in drug-induced apoptosis in human cancer cells. We propose that expression levels of DNMT1 in tumor cells may affect the effectiveness of doxorubicin in chemotherapy. PMID- 15340042 TI - Designing transcription factor architectures for drug discovery. AB - Recent advances in the design, selection, and engineering of DNA binding proteins have led to the emerging field of designer transcription factors (TFs). Modular DNA-binding protein domains can be assembled to recognize a given sequence of a DNA in a regulatory region of a targeted gene. TFs can be readily prepared by linking the DNA-binding protein to a variety of effector domains that mediate transcriptional activation or repression. Furthermore, the interaction between the TF and the genomic DNA can be regulated by several approaches, including chemical regulation by a variety of small molecules. Genome-wide single target specificity has been demonstrated using arrays of sequence-specific zinc finger (ZF) domains, polydactyl proteins. Any laboratory today can easily construct polydactyl ZF proteins by linkage of predefined ZF units that recognize specific triplets of DNA. The potential of this technology to alter the transcription of specific genes, to discover new genes, and to induce phenotypes in cells and organisms is now being applied in the areas of molecular therapeutics, pharmacology, biotechnology, and functional genomics. PMID- 15340043 TI - Mechanisms of agonist action at D2 dopamine receptors. AB - In this study, we investigated the biochemical mechanisms of agonist action at the G protein-coupled D2 dopamine receptor expressed in Chinese hamster ovary cells. Stimulation of guanosine 5'-O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) binding by full and partial agonists was determined at different concentrations of [35S]GTPgammaS (0.1 and 10 nM) and in the presence of different concentrations of GDP. At both concentrations of [35S]GTPgammaS, increasing GDP decreased the [35S]GTPgammaS binding observed with maximally stimulating concentrations of agonist, with partial agonists exhibiting greater sensitivity to the effects of GDP than full agonists. The relative efficacy of partial agonists was greater at the lower GDP concentrations. Concentration-response experiments were performed for a range of agonists at the two [35S]GTPgammaS concentrations and with different concentrations of GDP. At 0.1 nM [35S]GTPgammaS, the potency of both full and partial agonists was dependent on the GDP concentration in the assays. At 10 nM [35S]GTPgammaS, the potency of full agonists exhibited a greater dependence on the GDP concentration, whereas the potency of partial agonists was virtually independent of GDP. We concluded that at the lower [35S]GTPgammaS concentration, the rate-determining step in G protein activation is the binding of [35S]GTPgammaS to the G protein. At the higher [35S]GTPgammaS concentration, for full agonists, [35S]GTPgammaS binding remains the slowest step, whereas for partial agonists, another (GDP-independent) step, probably ternary complex breakdown, becomes rate-determining. PMID- 15340044 TI - Impaired CREB-1 phosphorylation in antifolate-resistant cell lines with down regulation of the reduced folate carrier gene. AB - The human reduced folate carrier (hRFC) is the dominant transporter for the uptake of antifolates used in cancer chemotherapy. We have shown recently that decreased cAMP-responsive element (CRE)-dependent transcription contributes to the loss of hRFC gene expression in multiple antifolate-resistant cell lines. This was associated with markedly decreased levels of phosphorylated cAMP response element-binding protein 1 (pCREB-1) and CRE-binding. Consistent with the autoregulation of CREB-1 gene expression by pCREB-1, prominently decreased CREB-1 mRNA levels were observed in antifolate-resistant cells. We therefore explored the possibility that these cells were defective in CREB-1 phosphorylation, thereby resulting in down-regulation of some cAMP-responsive genes. Two dimensional gel electrophoresis revealed that CREB-1 and its phosphoisoforms were markedly decreased in these cells. Treatment with forskolin, an activator of adenylyl cyclase, restored both CREB-1 and pCREB-1 levels; this resulted in the restoration of CRE-binding, CRE-reporter activity, and CREB-1 and RFC mRNA levels. Hence, the protein kinase A pathway was examined using various agents that augment intracellular cAMP levels, including cholera toxin, an upstream agonist that renders stimulatory G-proteins (Galphas) constitutively active. Treatment of antifolate-resistant cells with these agents resulted in the restoration of pCREB-1 levels and CRE-reporter activity. Furthermore, transient transfection with a constitutively transcriptionally active VP16-CREB-1 that does not require phosphorylation for its activity resulted in restoration of CREB mRNA levels but not pCREB-1 levels. This is the first demonstration that resistance to various antifolates may potentially be associated with impaired activity of Galphas or their coupled receptors, resulting in loss of CREB-1 phosphorylation and consequent down-regulation of cAMP-responsive genes. PMID- 15340045 TI - Critical role of reactive oxygen species and mitochondrial membrane potential in Korean mistletoe lectin-induced apoptosis in human hepatocarcinoma cells. AB - Viscum album L. coloratum agglutinin (VCA), isolated from Korean mistletoe, is a strong inducer of apoptosis in a variety of tumor cells; however, the underlying molecular mechanisms responsible are not clear. Here, we show that VCA induces apoptotic killing, as demonstrated by DNA fragmentation, Hoechst 33258 staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, and flow cytometry analysis in hepatocarcinoma Hep3B cells. VCA treatment results in a significant increase in reactive oxygen species (ROS) and loss of mitochondrial membrane potential (DeltaPsim). Furthermore, treatment with the antioxidant N acetyl-L-cysteine reduces ROS induction by VCA, preventing apoptosis in Hep3B cells, indicating that oxidative stress is involved in VCA-mediated cell death. Our results also show rapid changes in mitochondrial transition permeability, Bax translocation, cytochrome c release, caspase-3 activity, and poly(ADP-ribose) polymerase degradation in Hep3B cells occurring in VCA-induced apoptosis. There is much evidence that implicates c-Jun NH2-terminal kinase (JNK) activation with apoptosis in a variety of cellular and animal models. In this study, we show that VCA induces JNK phosphorylation, which is abolished with pretreatment with a JNK inhibitor. Moreover, Hep3B cells overexpressing JNK1 or stress-activated protein kinase kinase (SEK1) seem to be more susceptible to cell death from ROS and loss of DeltaPsim induced by VCA, whereas expression of dominant-negative JNK1 or SEK1 in Hep3B cells do not. These data suggest that JNK phosphorylation may be a major regulator involved in VCA-induced apoptosis. Together, these results suggest that VCA induces apoptosis by inducing ROS production and a loss of DeltaPsim, in which JNK phosphorylation plays a critical role in these events. PMID- 15340046 TI - Unexpectedly complex editing patterns at dinucleotide insertion sites in Physarum mitochondria. AB - Many of the RNAs transcribed from the mitochondrial genome of Physarum polycephalum are edited by the insertion of nonencoded nucleotides, which are added either singly or as dinucleotides. In addition, at least one mRNA is also subject to substitutional editing in which encoded C residues are changed to U residues posttranscriptionally. We have shown previously that the predominant type of editing in these organelles, the insertion of nonencoded single C residues, occurs cotranscriptionally at the growing end of the RNA chain. However, less is known about the timing of dinucleotide addition, and it has been suggested that these insertions occur at a later stage in RNA maturation. Here we examine the addition of both single nucleotides and dinucleotides into nascent RNAs synthesized in vitro and in vivo. The distribution of added nucleotides within individual cloned cDNAs supports the hypothesis that all insertion sites are processed at the same time relative to transcription. In addition, the patterns of partial editing and misediting observed within these nascent RNAs suggest that separate factors may be required at a subset of dinucleotide insertion sites and raise the possibility that in vivo, nucleotides may be added to RNA and then changed posttranscriptionally. PMID- 15340047 TI - RhoE inhibits cell cycle progression and Ras-induced transformation. AB - Rho GTPases are major regulators of cytoskeletal dynamics, but they also affect cell proliferation, transformation, and oncogenesis. RhoE, a member of the Rnd subfamily that does not detectably hydrolyze GTP, inhibits RhoA/ROCK signaling to promote actin stress fiber and focal adhesion disassembly. We have generated fibroblasts with inducible RhoE expression to investigate the role of RhoE in cell proliferation. RhoE expression induced a loss of stress fibers and cell rounding, but these effects were only transient. RhoE induction inhibited cell proliferation and serum-induced S-phase entry. Neither ROCK nor RhoA inhibition accounted for this response. Consistent with its inhibitory effect on cell cycle progression, RhoE expression was induced by cisplatin, a DNA damage-inducing agent. RhoE-expressing cells failed to accumulate cyclin D1 or p21(cip1) protein or to activate E2F-regulated genes in response to serum, although ERK, PI3-K/Akt, FAK, Rac, and cyclin D1 transcription was activated normally. The expression of proteins that bypass the retinoblastoma (pRb) family cell cycle checkpoint, including human papillomavirus E7, adenovirus E1A, and cyclin E, rescued cell cycle progression in RhoE-expressing cells. RhoE also inhibited Ras- and Raf induced fibroblast transformation. These results indicate that RhoE inhibits cell cycle progression upstream of the pRb checkpoint. PMID- 15340048 TI - A single pulse of agrin triggers a pathway that acts to cluster acetylcholine receptors. AB - Agrin triggers signaling mechanisms of high temporal and spatial specificity to achieve phosphorylation, clustering, and stabilization of postsynaptic acetylcholine receptors (AChRs). Agrin transiently activates the kinase MuSK; MuSK activation has largely vanished when AChR clusters appear. Thus, a tyrosine kinase cascade acts downstream from MuSK, as illustrated by the agrin-evoked long lasting activation of Src family kinases (SFKs) and their requirement for AChR cluster stabilization. We have investigated this cascade and report that pharmacological inhibition of SFKs reduces early but not later agrin-induced phosphorylation of MuSK and AChRs, while inhibition of Abl kinases reduces late phosphorylation. Interestingly, SFK inhibition applied selectively during agrin induced AChR cluster formation caused rapid cluster dispersal later upon agrin withdrawal. We also report that a single 5-min agrin pulse, followed by extensive washing, triggered long-lasting MuSK and AChR phosphorylation and efficient AChR clustering. Following the pulse, MuSK phosphorylation increased and, beyond a certain level, caused maximal clustering. These data reveal novel temporal aspects of tyrosine kinase action in agrin signaling. First, during AChR cluster formation, SFKs initiate early phosphorylation and an AChR stabilization program that acts much later. Second, a kinase mechanism rapidly activated by agrin acts thereafter autonomously in agrin's absence to further increase MuSK phosphorylation and cluster AChRs. PMID- 15340049 TI - An antisense RNA regulates the bidirectional silencing property of the Kcnq1 imprinting control region. AB - The Kcnq1 imprinting control region (ICR) located in intron 10 of the Kcnq1 gene is unmethylated on the paternal chromosome and methylated on the maternal chromosome and has been implicated in the manifestation of parent-of-origin specific expression of six neighboring genes. The unmethylated Kcnq1 ICR harbors bidirectional silencer activity and drives expression of an antisense RNA, Kcnq1ot1, which overlaps the Kcnq1 coding region. To elucidate whether the Kcnq1ot1 RNA plays a role in the bidirectional silencing activity of the Kcnq1 ICR, we have characterized factor binding sites by genomic footprinting and tested the functional consequence of various deletions of these binding sites in an episome-based system. Deletion of the elements necessary for Kcnq1ot1 promoter function resulted in the loss of silencing activity. Furthermore, interruption of Kcnq1ot1 RNA production by the insertion of a polyadenylation sequence downstream of the promoter also caused a loss of both silencing activity and methylation spreading. Thus, the antisense RNA plays a key role in the silencing function of the ICR. Double-stranded RNA (dsRNA)-mediated RNA interference is unlikely to be involved, as the ICR is active irrespective of the simultaneous production of dsRNA from the genes it silences. PMID- 15340050 TI - Transcription factor Nkx-2.5 induces sodium/iodide symporter gene expression and participates in retinoic acid- and lactation-induced transcription in mammary cells. AB - The sodium/iodide symporter (NIS) is a plasma membrane protein that mediates active iodide transport in thyroid and mammary cells. It is a prerequisite for radioiodide treatment of thyroid cancer and a promising diagnostic and therapeutic tool for breast cancer. We investigated the molecular mechanisms governing NIS expression in mammary cells. Here we report that Nkx-2.5, a cardiac homeobox transcription factor that is also expressed in the thyroid primordium, is a potent inducer of the NIS promoter. By binding to two specific promoter sites (N2 and W), Nkx-2.5 induced the rNIS promoter (about 50-fold over the basal level). Interestingly, coincident with NIS expression, Nkx-2.5 mRNA and protein were present in lactating, but not virgin, mammary glands in two human breast cancer samples and in all-trans retinoic acid (tRA)-stimulated MCF-7 breast cancer cells. A cotransfected dominant-negative Nkx-2.5 mutant abolished tRA induced endogenous NIS induction, which shows that Nkx-2.5 activity is critical for this process. Remarkably, in MCF-7 cells, Nkx-2.5 overexpression alone was sufficient to induce NIS and iodide uptake. In conclusion, Nkx-2.5 is a novel relevant transcriptional regulator of mammary NIS and could thus be exploited to manipulate NIS expression in breast cancer treatment strategies. PMID- 15340051 TI - A cellular RNA-binding protein enhances internal ribosomal entry site-dependent translation through an interaction downstream of the hepatitis C virus polyprotein initiation codon. AB - Translational initiation of hepatitis C virus (HCV) mRNA occurs by internal entry of ribosomes into an internal ribosomal entry site (IRES) at the 5' nontranslated region. A region encoding the N-terminal part of the HCV polyprotein has been shown to augment the translation of HCV mRNA. Here we show that a cellular protein, NS1-associated protein 1 (NSAP1), augments HCV mRNA translation through a specific interaction with an adenosine-rich protein-coding region within the HCV mRNA. The overexpression of NSAP1 specifically enhanced HCV IRES-dependent translation, and knockdown of NSAP1 by use of a small interfering RNA specifically inhibited the translation of HCV mRNA. An HCV replicon RNA capable of mimicking the HCV proliferation process in host cells was further used to confirm that NSAP1 enhances the translation of HCV mRNA. These results suggest the existence of a novel mechanism of translational enhancement that acts through the interaction of an RNA-binding protein with a protein coding sequence. PMID- 15340052 TI - Nature of the accessible chromatin at a glucocorticoid-responsive enhancer. AB - To gain a better understanding of the nature of active chromatin in mammals, we have characterized in living cells the various chromatin modification events triggered by the glucocorticoid receptor (GR) at the rat tyrosine aminotransferase gene. GR promotes a local remodeling at a glucocorticoid responsive unit (GRU) located 2.5 kb upstream of the transcription start site, creating nuclease hypersensitivity that encompasses 450 bp of DNA. Nucleosomes at the GRU occupy multiple frames that are remodeled without nucleosome repositioning, showing that nucleosome positioning is not the key determinant of chromatin accessibility at this locus. Remodeling affects nucleosomes and adjacent linker sequences, enhancing accessibility at both regions. This is associated with decreased interaction of both the linker histone H1 and the core histone H3 with DNA. Thus, our results indicate that nucleosome and linker histone removal rather than nucleosome repositioning is associated with GR triggered accessibility. Interestingly, GR induces hyperacetylation of histones H3 and H4, but this is not sufficient either for remodeling or for transcriptional activation. Finally, our data favor the coexistence of several chromatin states within the population, which may account for the previously encountered difficulties in characterizing unambiguously the active chromatin structure in living cells. PMID- 15340053 TI - Essential motifs in the 3' untranslated region required for retrotransposition and the precise start of reverse transcription in non-long-terminal-repeat retrotransposon SART1. AB - Non-long-terminal-repeat (non-LTR) retrotransposons amplify their copies by reverse transcribing mRNA from the 3' end, but the initial processes of reverse transcription are still unclear. We have shown that a telomere-specific non-LTR retrotransposon of the silkworm, SART1, requires the 3' untranslated region (3' UTR) for retrotransposition. With an in vivo retrotransposition assay, we identified several novel motifs within the 3' UTR involved in precise and efficient reverse transcription. Of 461 nucleotides (nt) of the 3' UTR, the central region, from nt 163 to nt 295, was essential for SART1 retrotransposition. Of five putative stem-loops formed in RNA for the SART1 3' UTR, the second stem-loop (nt 159 to 221) is included in this region. Loss of the 3' region (nt 296 to 461) in the 3' UTR and the poly(A) tract resulted in decreased and inaccurate reverse transcription, which starts mostly from several telomeric repeat-like GGUU sequences just downstream of the second stem-loop. These results suggest that short telomeric repeat-like sequences in the 3' UTR anneal to the bottom strand of (TTAGG)(n) repeats. We also demonstrated that the mRNA for green fluorescent protein (GFP) could be retrotransposed into telomeric repeats when the GFP coding region is fused with the SART1 3' UTR and SART1 open reading frame proteins are supplied in trans. PMID- 15340054 TI - Granulocyte-macrophage colony-stimulating factor enhancer activation requires cooperation between NFAT and AP-1 elements and is associated with extensive nucleosome reorganization. AB - The human granulocyte-macrophage colony-stimulating factor (GM-CSF) gene is activated by an NFAT-dependent enhancer forming an inducible DNase I hypersensitive (DH) site. The enhancer core comprising the DH site contains the GM330 and GM420 elements that bind NFAT and AP-1 cooperatively. Here we demonstrate that both elements are essential for enhancer activity and that Sp1 and AML1 sites in the enhancer become occupied in vivo only after activation. Chromatin structure analysis revealed that the GM-CSF enhancer core elements are divided between two adjacent nucleosomes that become destabilized and highly accessible after activation. Inducible chromatin reorganization was not restricted to the enhancer core but extended across a 3-kb domain of mobilized nucleosomes, within which the nucleosome repeat length was compressed from approximately 185 to 150 bp. The GM420 element is a high-affinity site that binds NFAT independently of AP-1 but depends on the linked AP-1 site for enhancer function. Nevertheless, just the NFAT motif from the GM420 element was sufficient to form a DH site within chromatin even in the absence of the AP-1 site. Hence, NFAT has the potential to cooperate with other transcription factors by promoting chromatin remodelling and increasing accessibility at inducible regulatory elements. PMID- 15340055 TI - Nuclear receptors CAR and PXR cross talk with FOXO1 to regulate genes that encode drug-metabolizing and gluconeogenic enzymes. AB - The nuclear receptors CAR and PXR activate hepatic genes in response to therapeutic drugs and xenobiotics, leading to the induction of drug-metabolizing enzymes, such as cytochrome P450. Insulin inhibits the ability of FOXO1 to express genes encoding gluconeogenic enzymes. Induction by drugs is known to be decreased by insulin, whereas gluconeogenic activity is often repressed by treatment with certain drugs, such as phenobarbital (PB). Performing cell-based transfection assays with drug-responsive and insulin-responsive enhancers, glutathione S-transferase pull down, RNA interference (RNAi), and mouse primary hepatocytes, we examined the molecular mechanism by which nuclear receptors and FOXO1 could coordinately regulate both enzyme pathways. FOXO1 was found to be a coactivator to CAR- and PXR-mediated transcription. In contrast, CAR and PXR, acting as corepressors, downregulated FOXO1-mediated transcription in the presence of their activators, such as 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP) and pregnenolone 16alpha-carbonitrile, respectively. A constitutively active mutant of the insulin-responsive protein kinase Akt, but not the kinase negative mutant, effectively blocked FOXO1 activity in cell-based assays. Thus, insulin could repress the receptors by activating the Akt-FOXO1 signal, whereas drugs could interfere with FOXO1-mediated transcription by activating CAR and/or PXR. Treatment with TCPOBOP or PB decreased the levels of phosphoenolpyruvate carboxykinase 1 mRNA in mice but not in Car(-/-) mice. We conclude that FOXO1 and the nuclear receptors reciprocally coregulate their target genes, modulating both drug metabolism and gluconeogenesis. PMID- 15340056 TI - Different effects of CSA and CSB deficiency on sensitivity to oxidative DNA damage. AB - Mutations in the CSA and CSB genes cause Cockayne syndrome, a rare inherited disorder characterized by UV sensitivity, severe neurological abnormalities, and progeriod symptoms. Both gene products function in the transcription-coupled repair (TCR) subpathway of nucleotide excision repair (NER), providing the cell with a mechanism to remove transcription-blocking lesions from the transcribed strands of actively transcribed genes. Besides a function in TCR of NER lesions, a role of CSB in (transcription-coupled) repair of oxidative DNA damage has been suggested. In this study we used mouse models to compare the effect of a CSA or a CSB defect on oxidative DNA damage sensitivity at the levels of the cell and the intact organism. In contrast to CSB(-/-) mouse embryonic fibroblasts (MEFs), CSA( /-) MEFs are not hypersensitive to gamma-ray or paraquat treatment. Similar results were obtained for keratinocytes. In contrast, both CSB(-/-) and CSA(-/-) embryonic stem cells show slight gamma-ray sensitivity. Finally, CSB(-/-) but not CSA(-/-) mice fed with food containing di(2-ethylhexyl)phthalate (causing elevated levels of oxidative DNA damage in the liver) show weight reduction. These findings not only uncover a clear difference in oxidative DNA damage sensitivity between CSA- and CSB-deficient cell lines and mice but also show that sensitivity to oxidative DNA damage is not a uniform characteristic of Cockayne syndrome. This difference in the DNA damage response between CSA- and CSB deficient cells is unexpected, since until now no consistent differences between CSA and CSB patients have been reported. We suggest that the CSA and CSB proteins in part perform separate roles in different DNA damage response pathways. PMID- 15340057 TI - Innate immune responses in peptidoglycan recognition protein L-deficient mice. AB - Peptidoglycan recognition proteins (PGRPs) constitute a family of innate immune recognition molecules. In Drosophila, distinct PGRPs bind to peptidoglycans on gram-positive or gram-negative bacteria and provide essential signals upstream of the Toll and Imd pathways required for immunity against infection. Four PGRPs, PGRP-L, -S, -Ialpha, and -Ibeta, are expressed from three genes in mammals. In this paper, we provide direct evidence that the longest family member, PGRP-L, is a secreted serum protein with the capacity to multimerize. Using gene targeting to create PGRP-L-deficient mice, we demonstrate little contribution by PGRP-L to systemic challenge using gram-negative bacteria (Escherichia coli, slightly less susceptible), Gram-positive bacteria (Staphylococcus aureus), or yeast (Candida albicans). Peritoneal macrophages from PGRP-L-deficient mice produced decreased amounts of the inflammatory cytokines interleukin 6 and tumor necrosis factor alpha when stimulated with E. coli or lipopolysaccharide, but comparable amounts when stimulated with S. aureus, C. albicans, or their cell wall components. Additionally, these cells produced similar amounts of cytokines when challenged with gram-positive or -negative peptidoglycans. In contrast to its critical role in immunity in flies, PGRP-L is largely dispensable for mammalian immunity against bacteria and fungi. PMID- 15340058 TI - Mice deficient in the axonemal protein Tektin-t exhibit male infertility and immotile-cilium syndrome due to impaired inner arm dynein function. AB - The haploid germ cell-specific Tektin-t protein is a member of the Tektin family of proteins that form filaments in flagellar, ciliary, and axonemal microtubules. To investigate the physiological role of Tektin-t, we generated mice with a mutation in the tektin-t gene. The homozygous mutant males were infertile, while the females were fully fertile. Sperm morphology and function were abnormal, with frequent bending of the sperm flagella and marked defects in motility. In vitro fertilization assays showed that the defective spermatozoa were able to fertilize eggs. Electron microscopic examination showed that the dynein inner arm structure was disrupted in the sperm flagella of tektin-t-deficient mice. Furthermore, homozygous mutant mice had functionally defective tracheal cilia, as evidenced by altered dynein arm morphology. These results indicate that Tektin-t participates in dynein inner arm formation or attachment and that the loss of Tektin-t results in impaired motility of both flagella and cilia. Therefore, the tektin-t gene is one of the causal genes for immotile-cilium syndrome/primary ciliary dyskinesia. PMID- 15340059 TI - Biochemical and functional characterizations of small GTPase Rheb and TSC2 GAP activity. AB - Tuberous sclerosis complex (TSC) is a genetic disease caused by a mutation in either the tsc1 or tsc2 tumor suppressor gene. Recent studies have demonstrated that TSC2 displays GAP (GTPase-activating protein) activity specifically towards the small G protein Rheb and inhibits its ability to stimulate the mTOR signaling pathway. Rheb and TSC2 comprise a unique pair of GTPase and GAP, because Rheb has high basal GTP levels and TSC2 does not have the catalytic arginine finger found in Ras-GAP. To investigate the function of TSC2 and Rheb in mTOR signaling, we analyzed the TSC2-stimulated Rheb GTPase activity. We found that Arg15, a residue equivalent to Gly12 in Ras, is important for Rheb to function as a substrate for TSC2 GAP. In addition, we identified asparagine residues essential for TSC2 GAP activity. We demonstrated a novel catalytic mechanism of the TSC2 GAP and Rheb that TSC2 uses a catalytic "asparagine thumb" instead of the arginine finger found in Ras-GAP. Furthermore, we discovered that farnesylation and membrane localization of Rheb is not essential for Rheb to stimulate S6 kinase (S6K) phosphorylation. Analysis of TSC1 binding defective mutants of TSC2 shows that TSC1 is not required for the TSC2 GAP activity but may function as a regulatory component in the TSC1/TSC2 complex. Our data further demonstrate that GAP activity is essential for the cellular function of TSC2 to inhibit S6K phosphorylation. PMID- 15340060 TI - The small nucle(ol)ar RNA cap trimethyltransferase is required for ribosome synthesis and intact nucleolar morphology. AB - Nucleolar morphogenesis is a poorly defined process. Here we report that the Saccharomyces cerevisiae nucleolar trimethyl guanosine synthase I (Tgs1p), which specifically selects the m(7)G cap structure of snRNAs and snoRNAs for m(2,2,7)G conversion, is required not only for efficient pre-mRNA splicing but also for pre rRNA processing and small ribosomal subunit synthesis. Mutational analysis indicates that the requirement for Tgs1p in pre-mRNA splicing, but not its involvement in ribosome synthesis, is dependent upon its function in cap trimethylation. In addition, we report that cells lacking Tgs1p showed a striking and unexpected loss of nucleolar structural organization. Tgs1p is not a core component of the snoRNP proteins; however, in vitro, the protein interacts with the KKD/E domain present at the carboxyl-terminal ends of several snoRNP proteins. Strains expressing versions of the snoRNPs lacking the KKD/E domain were also defective for nucleolar morphology and showed a loss of nucleolar compaction. We propose that the transient and functional interactions of Tgs1p with the abundant snoRNPs, through presumed interactions with the KKD/E domain of the snoRNP proteins, contribute substantially to the coalescence of nucleolar components. This conclusion is compatible with a model of self-organization for nucleolar assembly. PMID- 15340061 TI - Regulation of human p53 activity and cell localization by alternative splicing. AB - The development of cancer is a multistep process involving mutations in proto oncogenes, tumor suppressor genes, and other genes which control cell proliferation, telomere stability, angiogenesis, and other complex traits. Despite this complexity, the cellular pathways controlled by the p53 tumor suppressor protein are compromised in most, if not all, cancers. In normal cells, p53 controls cell proliferation, senescence, and/or mediates apoptosis in response to stress, cell damage, or ectopic oncogene expression, properties which make p53 the prototype tumor suppressor gene. Defining the mechanisms of regulation of p53 activity in normal and tumor cells has therefore been a major priority in cell biology and cancer research. The present study reveals a novel and potent mechanism of p53 regulation originating through alternative splicing of the human p53 gene resulting in the expression of a novel p53 mRNA. This novel p53 mRNA encodes an N-terminally deleted isoform of p53 termed p47. As demonstrated within, p47 was able to effectively suppress p53-mediated transcriptional activity and impair p53-mediated growth suppression. It was possible to select for p53-null cells expressing p47 alone or coexpressing p53 in the presence of p47 but not cells expressing p53 alone. This showed that p47 itself does not suppress cell viability but could control p53-mediated growth suppression. Interestingly, p47 was monoubiquitinated in an Mdm2-independent manner, and this was associated with its export out of the nucleus. In the presence of p47, there was a reduction in Mdm2-mediated polyubiquitination and degradation of p53, and this was also associated with increased monoubiquitination and nuclear export of p53. The expression of p47 through alternative splicing of the p53 gene thus has a major influence over p53 activity at least in part through controlling p53 ubiquitination and cell localization. PMID- 15340062 TI - Targeted gene knockout reveals a role in meiotic recombination for ZHP-3, a Zip3 related protein in Caenorhabditis elegans. AB - The meiotically expressed Zip3 protein is found conserved from Saccharomyces cerevisiae to humans. In baker's yeast, Zip3p has been implicated in synaptonemal complex (SC) formation, while little is known about the protein's function in multicellular organisms. We report here the successful targeted gene disruption of zhp-3 (K02B12.8), the ZIP3 homolog in the nematode Caenorhabditis elegans. Homozygous zhp-3 knockout worms show normal homologue pairing and SC formation. Also, the timing of appearance and the nuclear localization of the recombination protein Rad-51 seem normal in these animals, suggesting proper initiation of meiotic recombination by DNA double-strand breaks. However, the occurrence of univalents during diplotene indicates that C. elegans ZHP-3 protein is essential for reciprocal recombination between homologous chromosomes and thus chiasma formation. In the absence of ZHP-3, reciprocal recombination is abolished and double-strand breaks seem to be repaired via alternative pathways, leading to achiasmatic chromosomes and the occurrence of univalents during meiosis I. Green fluorescent protein-tagged C. elegans ZHP-3 forms lines between synapsed chromosomes and requires the SC for its proper localization. PMID- 15340063 TI - Dilated cardiomyopathy caused by aberrant endoplasmic reticulum quality control in mutant KDEL receptor transgenic mice. AB - Aberrant protein folding beyond the capacity of endoplasmic reticulum (ER) quality control leads to stress response in the ER. The Lys-Asp-Glu-Leu (KDEL) receptor, a retrieval receptor for ER chaperones in the early secretory pathway, contributes to ER quality control. To elucidate the function of the KDEL receptor in vivo, we established transgenic mice expressing a mutant KDEL receptor. We found that the mutant KDEL receptor sensitized cells to ER stress and that the mutant mice developed dilated cardiomyopathy. Ultrastructural analyses revealed expanded sarcoplasmic reticulums and protein aggregates that obstructed the adjacent transverse tubules of the mutant cardiomyocytes. Cardiomyocytes from the mutant mice were sensitive to ER stress when treated with tunicamycin and showed a functional defect in the L-type Ca(2+) current. We observed ubiquitinated protein aggregates, enhanced expression of CHOP (a death-related transcriptional factor expressed upon ER stress), and apoptosis in the mutant hearts. These findings suggest that impairment of the KDEL receptor disturbs ER quality control, resulting in accumulation of misfolded proteins in the ER in an in vivo system, and that the dilated cardiomyopathy found in the mutant KDEL receptor transgenic mice is associated with ER stress. PMID- 15340064 TI - Activation of REST/NRSF target genes in neural stem cells is sufficient to cause neuronal differentiation. AB - REST/NRSF is a transcriptional repressor that acts at the terminal stage of the neuronal differentiation pathway and blocks the transcription of several differentiation genes. REST/NRSF is generally downregulated during induction of neuronal differentiation. The recombinant transcription factor REST-VP16 binds to the same DNA binding site as does REST/NRSF but functions as an activator instead of a repressor and can directly activate the transcription of REST/NRSF target genes. However, it is not known whether REST-VP16 expression is sufficient to cause formation of functional neurons from neural stem cells (NSCs). Here we show that regulated expression of REST-VP16 in a physiologically relevant NSC line growing under cycling conditions converted the cells rapidly to the mature neuronal phenotype. Furthermore, when grown in the presence of retinoic acid, REST-VP16-expressing NSCs activated their target, as well as other differentiation genes that are not their direct target, converting them to the mature neuronal phenotype and enabling them to survive in the presence of mitotic inhibitors, which is a characteristic of mature neurons. In addition, these neuronal cells were physiologically active. These results showed that direct activation of REST/NRSF target genes in NSCs with a single transgene, REST-VP16, is sufficient to cause neuronal differentiation, and the findings suggested that direct activation of genes involved in the terminal stage of differentiation may cause neuronal differentiation of NSCs. PMID- 15340065 TI - A targeting mutation of tyrosine 1062 in Ret causes a marked decrease of enteric neurons and renal hypoplasia. AB - The Ret receptor tyrosine kinase plays a crucial role in the development of the enteric nervous system and the kidney. Tyrosine 1062 in Ret represents a binding site for the phosphotyrosine-binding domains of several adaptor and effector proteins that are important for the activation of intracellular signaling pathways, such as the RAS/ERK, phosphatidylinositol 3-kinase/AKT, and Jun associated N-terminal kinase pathways. To investigate the importance of tyrosine 1062 for organogenesis in vivo, knock-in mice in which tyrosine 1062 in Ret was replaced with phenylalanine were generated. Although homozygous knock-in mice were born normally, they died by day 27 after birth and showed growth retardation. The development of the enteric nervous system was severely impaired in homozygous mutant mice, about 40% of which lacked enteric neurons in the whole intestinal tract, as observed in Ret-deficient mice. The rest of the mutant mice developed enteric neurons in the intestine to various extents, although the size and number of ganglion cells were significantly reduced. Unlike Ret-deficient mice, a small kidney developed in all knock-in mice, accompanying a slight histological change. The reduction of kidney size was due to a decrease of ureteric bud branching during embryogenesis. Thus, these findings demonstrated that the signal via tyrosine 1062 plays an important role in histogenesis of the enteric nervous system and nephrogenesis. PMID- 15340066 TI - Inactivation of Stat5 in mouse mammary epithelium during pregnancy reveals distinct functions in cell proliferation, survival, and differentiation. AB - This study explored the functions of the signal transducers and activators of transcription 5a and 5b (referred to as Stat5 here) during different stages of mouse mammary gland development by using conditional gene inactivation. Mammary gland morphogenesis includes cell specification, proliferation and differentiation during pregnancy, cell survival and maintenance of differentiation throughout lactation, and cell death during involution. Stat5 is activated by prolactin, and its presence is mandatory for the proliferation and differentiation of mammary epithelium during pregnancy. To address the question of whether Stat5 is also necessary for the maintenance and survival of the differentiated epithelium, the two genes were deleted at different time points. The 110-kb Stat5 locus in the mouse was bracketed with loxP sites, and its deletion was accomplished by using two Cre-expressing transgenic lines. Loss of Stat5 prior to pregnancy prevented epithelial proliferation and differentiation. Deletion of Stat5 during pregnancy, after mammary epithelium had entered Stat5 mediated differentiation, resulted in premature cell death, indicating that at this stage epithelial cell proliferation, differentiation, and survival require Stat5. PMID- 15340067 TI - Heterotrimeric G proteins of the Gq/11 family are crucial for the induction of maternal behavior in mice. AB - Heterotrimeric G proteins of the G(q/11) family transduce signals from a variety of neurotransmitter receptors and have therefore been implicated in several functions of the central nervous system. To investigate the potential role of G(q/11) signaling in behavior, we generated mice which lack the alpha-subunits of the two main members of the G(q/11) family, Galpha(q) and Galpha(11), selectively in the forebrain. We show here that forebrain Galpha(q/11)-deficient females do not display any maternal behavior such as nest building, pup retrieving, crouching, or nursing. However, olfaction, motor behavior and mammary gland function are normal in forebrain Galpha(q/11)-deficient females. We used c-fos immunohistochemistry to investigate pup-induced neuronal activation in different forebrain regions and found a significant reduction in the medial preoptic area, the bed nucleus of stria terminalis, and the lateral septum both in postpartum females and in virgin females after foster pup exposure. Pituitary function, especially prolactin release, was normal in forebrain Galpha(q/11)-deficient females, and activation of oxytocin receptor-positive neurons in the hypothalamus did not differ between genotypes. Our findings show that G(q/11) signaling is indispensable to the neuronal circuit that connects the perception of pup-related stimuli to the initiation of maternal behavior and that this defect cannot be attributed to either reduced systemic prolactin levels or impaired activation of oxytocin receptor-positive neurons of the hypothalamus. PMID- 15340068 TI - Sequestosome 1/p62 is a polyubiquitin chain binding protein involved in ubiquitin proteasome degradation. AB - Herein, we demonstrate that the ubiquitin-associated (UBA) domain of sequestosome 1/p62 displays a preference for binding K63-polyubiquitinated substrates. Furthermore, the UBA domain of p62 was necessary for aggregate sequestration and cell survival. However, the inhibition of proteasome function compromised survival in cells with aggregates. Mutational analysis of the UBA domain reveals that the conserved hydrophobic patch MGF as well as the conserved leucine in helix 2 are necessary for binding polyubiquitinated proteins and for sequestration-aggregate formation. We report that p62 interacts with the proteasome by pull-down assay, coimmunoprecipitation, and colocalization. Depletion of p62 levels results in an inhibition of ubiquitin proteasome-mediated degradation and an accumulation of ubiquitinated proteins. Altogether, our results support the hypothesis that p62 may act as a critical ubiquitin chain targeting factor that shuttles substrates for proteasomal degradation. PMID- 15340069 TI - Sgt1 associates with Hsp90: an initial step of assembly of the core kinetochore complex. AB - The kinetochore, which consists of DNA sequence elements and structural proteins, is essential for high-fidelity chromosome transmission during cell division. In budding yeast, Sgt1, together with Skp1, is required for assembly of the core kinetochore complex (CBF3) via Ctf13 activation. Formation of the active Ctf13 Skp1 complex also requires Hsp90, a molecular chaperone. We have found that Sgt1 interacts with Hsp90 in yeast. We also have determined that Skp1 and Hsc82 (a yeast Hsp90 protein) bind to the N-terminal region of Sgt1 that contains tetratricopeptide repeat motifs. Results of sequence and phenotypic analyses of sgt1 mutants strongly suggest that the N-terminal region containing the Hsc82 binding and Skp1-binding domains of Sgt1 is important for the kinetochore function of Sgt1. We found that Hsp90's binding to Sgt1 stimulates the binding of Sgt1 to Skp1 and that Sgt1 and Hsp90 stimulate the binding of Skp1 to Ctf13, the F-box core kinetochore protein. Our results strongly suggest that Sgt1 and Hsp90 function in assembling CBF3 by activating Skp1 and Ctf13. PMID- 15340070 TI - Drosophila Ada2b is required for viability and normal histone H3 acetylation. AB - Regulation of chromatin through histone acetylation is an important step in gene expression. The Gcn5 histone acetyltransferase is part of protein complexes, e.g., the SAGA complex, that interact with transcriptional activators, targeting the enzyme to specific promoters and assisting in recruitment of the basal RNA polymerase transcription machinery. The Ada2 protein directly binds to Gcn5 and stimulates its catalytic activity. Drosophila contains two Ada2 proteins, Drosophila Ada2a (dAda2a) and dAda2b. We have generated flies that lack dAda2b, which is part of a Drosophila SAGA-like complex. dAda2b is required for viability in Drosophila, and its deletion causes a reduction in histone H3 acetylation. A global hypoacetylation of chromatin was detected on polytene chromosomes in dAda2b mutants. This indicates that the dGcn5-dAda2b complex could have functions in addition to assisting in transcriptional activation through gene-specific acetylation. Although the Drosophila p53 protein was previously shown to interact with the SAGA-like complex in vitro, we find that p53 induction of reaper gene expression occurs normally in dAda2b mutants. Moreover, dAda2b mutant animals show excessive p53-dependent apoptosis in response to gamma radiation. Based on this result, we speculate that dAda2b may be necessary for efficient DNA repair or generation of a DNA damage signal. This could be an evolutionarily conserved function, since a yeast ada2 mutant is also sensitive to a genotoxic agent. PMID- 15340071 TI - Sequential histone modifications at Hoxd4 regulatory regions distinguish anterior from posterior embryonic compartments. AB - Hox genes are differentially expressed along the embryonic anteroposterior axis. We used chromatin immunoprecipitation to detect chromatin changes at the Hoxd4 locus during neurogenesis in P19 cells and embryonic day 8.0 (E8.0) and E10.5 mouse embryos. During Hoxd4 induction in both systems, we observed that histone modifications typical of transcriptionally active chromatin occurred first at the 3' neural enhancer and then at the promoter. Moreover, the sequential distribution of histone modifications between E8.0 and E10.5 was consistent with a spreading of open chromatin, starting with the enhancer, followed by successively more 5' intervening sequences, and culminating at the promoter. Neither RNA polymerase II (Pol II) nor CBP associated with the inactive gene. During Hoxd4 induction, CBP and RNA Pol II were recruited first to the enhancer and then to the promoter. Whereas the CBP association was transient, RNA Pol II remained associated with both regulatory regions. Histone modification and transcription factor recruitment occurred in posterior, Hox-expressing embryonic tissues, but never in anterior tissues, where such genes are inactive. Together, our observations demonstrate that the direction of histone modifications at Hoxd4 mirrors colinear gene activation across Hox clusters and that the establishment of anterior and posterior compartments is accompanied by the imposition of distinct chromatin states. PMID- 15340072 TI - Global role of TATA box-binding protein recruitment to promoters in mediating gene expression profiles. AB - The recruitment of TATA box-binding protein (TBP) to promoters is one of the rate limiting steps during transcription initiation. However, the global importance of TBP recruitment in determining the absolute and changing levels of transcription across the genome is not known. We used a genomic approach to explore the relationship between TBP recruitment to promoters and global gene expression profiles in Saccharomyces cerevisiae. Our data indicate that first, RNA polymerase III promoters are the most prominent binding targets of TBP in vivo. Second, the steady-state transcript levels of genes throughout the genome are proportional to the occupancy of their promoters by TBP, and changes in the expression levels of these genes are closely correlated with changes in TBP recruitment to their promoters. Third, a consensus TATA element does not appear to be a major determinant of either TBP binding or gene expression throughout the genome. Our results indicate that the recruitment of TBP to promoters in vivo is of universal importance in determining gene expression levels in yeast, regardless of the nature of the core promoter or the type of activator or repressor that may mediate changes in transcription. The primary data reported here are available at http://www.iyerlab.org/tbp. PMID- 15340073 TI - SRC-mediated phosphorylation of focal adhesion kinase couples actin and adhesion dynamics to survival signaling. AB - Integrin-associated focal adhesions not only provide adhesive links between cellular actin and extracellular matrix but also are sites of signal transmission into the cell interior. Many cell responses signal through focal adhesion kinase (FAK), often by integrin-induced autophosphorylation of FAK or phosphorylation by Src family kinases. Here, we used an interfering FAK mutant (4-9F-FAK) to show that Src-dependent FAK phosphorylation is required for focal adhesion turnover and cell migration, by controlling assembly of a calpain 2/FAK/Src/p42ERK complex, calpain activation, and proteolysis of FAK. Expression of 4-9F-FAK in FAK-deficient fibroblasts also disrupts F-actin assembly associated with normal adhesion and spreading. In addition, we found that FAK's ability to regulate both assembly and disassembly of the actin and adhesion networks may be linked to regulation of the protease calpain. Surprisingly, we also found that the same interfering 4-9F-FAK mutant protein causes apoptosis of serum-deprived, transformed cells and suppresses anchorage-independent growth. These data show that Src-mediated phosphorylation of FAK acts as a pivotal regulator of both actin and adhesion dynamics and survival signaling, which, in turn, control apparently distinct processes such as cell migration and anchorage-independent growth. This also highlights that dynamic regulation of actin and adhesions (which include the integrin matrix receptors) is critical to signaling output and biological responses. PMID- 15340074 TI - Integrin-linked kinase regulates endothelial cell survival and vascular development. AB - Integrin-linked kinase (ILK) is a phosphoinositide 3-kinase-dependent serine/threonine kinase that interacts with beta integrins. Here we show that endothelial cell (EC)-specific deletion of ILK in mice confers placental insufficiency with decreased labyrinthine vascularization, yielding no viable offspring. Deletion of ILK in zebra fish using antisense morpholino oligonucleotides results in marked patterning abnormalities of the vasculature and is similarly lethal. To dissect potential mechanisms responsible for these phenotypes, we performed ex vivo deletion of ILK from purified EC of adult mice. We observed downregulation of the active-conformation of beta1 integrins with a striking increase in EC apoptosis associated with activation of caspase 9. There was also reduced phosphorylation of the ILK kinase substrate, Akt. However, phenotypic rescue of ILK-deficient EC by wild-type ILK, but not by a constitutively active mutant of Akt, suggests regulation of EC survival by ILK in an Akt-independent manner. Thus, endothelial ILK plays a critical role in vascular development through integrin-matrix interactions and EC survival. These data have important implications for both physiological and pathological angiogenesis. PMID- 15340075 TI - Spontaneous mutagenesis is enhanced in Apex heterozygous mice. AB - Germ line DNA directs the development of the next generation and, as such, is profoundly different from somatic cell DNA. Spermatogenic cells obtained from young adult lacI transgenic mice display a lower spontaneous mutant frequency and greater in vitro base excision repair activity than somatic cells and tissues obtained from the same mice. However, spermatogenic cells from old lacI mice display a 10-fold higher mutant frequency. This increased spontaneous mutant frequency occurs coincidentally with decreased in vitro base excision repair activity for germ cell and testicular extracts that in turn corresponds to a decreased abundance of AP endonuclease. To directly test whether a genetic diminution of AP endonuclease results in increased spontaneous mutant frequencies in spermatogenic cell types, AP endonuclease heterozygous (Apex(+/-)) knockout mice were crossed with lacI transgenic mice. Spontaneous mutant frequencies were significantly elevated (approximately twofold) for liver and spleen obtained from 3-month-old Apex(+/-) lacI(+) mice compared to frequencies from Apex(+/+) lacI(+) littermates and were additionally elevated for somatic tissues from 9-month-old mice. Spermatogenic cells from 9-month-old Apex(+/-) lacI(+) mice were significantly elevated twofold compared to levels for 9-month-old Apex(+/+) lacI(+) control mice. These data indicate that diminution of AP endonuclease has a significant effect on spontaneous mutagenesis in somatic and germ line cells. PMID- 15340076 TI - Transcriptional regulation of human CYP27 integrates retinoid, peroxisome proliferator-activated receptor, and liver X receptor signaling in macrophages. AB - Cholesterol uptake and efflux are key metabolic processes associated with macrophage physiology and atherosclerosis. Peroxisome proliferator-activated receptor gamma (PPARgamma) and liver X receptor alpha (LXRalpha) have been linked to the regulation of these processes. It remains to be identified how activation of these receptors is connected and regulated by endogenous lipid molecules. We identified CYP27, a p450 enzyme, as a link between retinoid, PPARgamma, and LXR signaling. We show that the human CYP27 gene is under coupled regulation by retinoids and ligands of PPARs via a PPAR-retinoic acid receptor response element in its promoter. Induction of the enzyme's expression results in an increased level of 27-hydroxycholesterol and upregulation of LXR-mediated processes. Upregulated CYP27 activity also leads to LXR-independent elimination of CYP27 metabolites as an alternative means of cholesterol efflux. Moreover, human macrophage-rich atherosclerotic lesions have an increased level of retinoid-, PPARgamma-, and LXR-regulated gene expression and also enhanced CYP27 levels. Our findings suggest that nuclear receptor-regulated CYP27 expression is likely to be a key integrator of retinoic acid receptor-PPARgamma-LXR signaling, relying on natural ligands and contributing to lipid metabolism in macrophages. PMID- 15340077 TI - Protein kinase Cdelta regulates keratinocyte death and survival by regulating activity and subcellular localization of a p38delta-extracellular signal regulated kinase 1/2 complex. AB - Protein kinase Cdelta (PKCdelta) is an important regulator of apoptosis in epidermal keratinocytes. However, little information is available regarding the downstream kinases that mediate PKCdelta-dependent keratinocyte death. This study implicates p38delta mitogen-activated protein kinase (MAPK) as a downstream carrier of the PKCdelta-dependent death signal. We show that coexpression of PKCdelta with p38delta produces profound apoptosis-like morphological changes. These morphological changes are associated with increased sub-G(1) cell population, cytochrome c release, loss of mitochondrial membrane potential, caspase activation, and PARP cleavage. This death response is specific for the combination of PKCdelta and p38delta and is not produced by replacing PKCdelta with PKCalpha or p38delta with p38alpha. A constitutively active form of MEK6, an upstream activator of p38delta, can also produce cell death when coupled with p38delta. In addition, concurrent p38delta activation and extracellular signal regulated kinase 1/2 (ERK1/2) inactivation are required for apoptosis. Regarding this inverse regulation, we describe a p38delta-ERK1/2 complex that may coordinate these changes in activity. We further show that this p38delta-ERK1/2 complex relocates into the nucleus in response to PKCdelta expression. This regulation appears to be physiological, since H(2)O(2), a known inducer of keratinocyte apoptosis, promotes identical PKCdelta and p38delta-ERK1/2 activity changes, leading to similar morphological changes. PMID- 15340078 TI - Role of F-box protein betaTrcp1 in mammary gland development and tumorigenesis. AB - The F-box protein betaTrcp1 controls the stability of several crucial regulators of proliferation and apoptosis, including certain inhibitors of the NF-kappaB family of transcription factors. Here we show that mammary glands of betaTrcp1(-/ ) female mice display a hypoplastic phenotype, whereas no effects on cell proliferation are observed in other somatic cells. To investigate further the role of betaTrcp1 in mammary gland development, we generated transgenic mice expressing human betaTrcp1 targeted to epithelial cells under the control of the mouse mammary tumor virus (MMTV) long terminal repeat promoter. Compared to controls, MMTV betaTrcp1 mammary glands display an increase in lateral ductal branching and extensive arrays of alveolus-like protuberances. The mammary epithelia of MMTV betaTrcp1 mice proliferate more and show increased NF-kappaB DNA binding activity and higher levels of nuclear NF-kappaB p65/RelA. In addition, 38% of transgenic mice develop tumors, including mammary, ovarian, and uterine carcinomas. The targeting of betaTrcp1 to lymphoid organs produces no effects on these tissues. In summary, our results support the notion that betaTrcp1 positively controls the proliferation of breast epithelium and indicate that alteration of betaTrcp1 function and expression may contribute to malignant behavior of breast tumors, at least in part through NF-kappaB transactivation. PMID- 15340079 TI - Mutant huntingtin impairs axonal trafficking in mammalian neurons in vivo and in vitro. AB - Recent data in invertebrates demonstrated that huntingtin (htt) is essential for fast axonal trafficking. Here, we provide direct and functional evidence that htt is involved in fast axonal trafficking in mammals. Moreover, expression of full length mutant htt (mhtt) impairs vesicular and mitochondrial trafficking in mammalian neurons in vitro and in whole animals in vivo. Particularly, mitochondria become progressively immobilized and stop more frequently in neurons from transgenic animals. These defects occurred early in development prior to the onset of measurable neurological or mitochondrial abnormalities. Consistent with a progressive loss of function, wild-type htt, trafficking motors, and mitochondrial components were selectively sequestered by mhtt in human Huntington's disease-affected brain. Data provide a model for how loss of htt function causes toxicity; mhtt-mediated aggregation sequesters htt and components of trafficking machinery leading to loss of mitochondrial motility and eventual mitochondrial dysfunction. PMID- 15340080 TI - cis-Acting determinants of heterochromatin formation on Drosophila melanogaster chromosome four. AB - The heterochromatic domains of Drosophila melanogaster (pericentric heterochromatin, telomeres, and the fourth chromosome) are characterized by histone hypoacetylation, high levels of histone H3 methylated on lysine 9 (H3 mK9), and association with heterochromatin protein 1 (HP1). While the specific interaction of HP1 with both H3-mK9 and histone methyltransferases suggests a mechanism for the maintenance of heterochromatin, it leaves open the question of how heterochromatin formation is targeted to specific domains. Expression characteristics of reporter transgenes inserted at different sites in the fourth chromosome define a minimum of three euchromatic and three heterochromatic domains, interspersed. Here we searched for cis-acting DNA sequence determinants that specify heterochromatic domains. Genetic screens for a switch in phenotype demonstrate that local deletions or duplications of 5 to 80 kb of DNA flanking a transposon reporter can lead to the loss or acquisition of variegation, pointing to short-range cis-acting determinants for silencing. This silencing is dependent on HP1. A switch in transgene expression correlates with a switch in chromatin structure, judged by nuclease accessibility. Mapping data implicate the 1360 transposon as a target for heterochromatin formation. We propose that heterochromatin formation is initiated at dispersed repetitive elements along the fourth chromosome and spreads for approximately 10 kb or until encountering competition from a euchromatic determinant. PMID- 15340081 TI - Generation and characterization of neuregulin-2-deficient mice. AB - The neuregulins (NRGs) are a family of four structurally related growth factors that are expressed in the developing and adult brain. NRG-1 is essential for normal heart formation and has been implicated in the development and maintenance of both neurons and glia. NRG-2 was identified on the basis of its homology to NRG-1 and, like NRG-1, is expressed predominantly by neurons in the central nervous system. We have generated mice with the active domain of NRG-2 deleted in an effort to characterize the biological function of NRG-2 in vivo. In contrast to the NRG-1 knockout animals, NRG-2 knockouts have no apparent heart defects and survive embryogenesis. Mutant mice display early growth retardation and reduced reproductive capacity. No obvious histological differences were observed in the major sites of NRG-2 expression. Our results indicate that in vivo NRG-2 activity differs substantially from that of NRG-1 and that it is not essential for normal development in utero. PMID- 15340082 TI - The Swi/Snf chromatin remodeling complex is required for ribosomal DNA and telomeric silencing in Saccharomyces cerevisiae. AB - The Swi/Snf chromatin remodeling complex has been previously demonstrated to be required for transcriptional activation and repression of a subset of genes in Saccharomyces cerevisiae. In this work we demonstrate that Swi/Snf is also required for repression of RNA polymerase II-dependent transcription in the ribosomal DNA (rDNA) locus (rDNA silencing). This repression appears to be independent of both Sir2 and Set1, two factors known to be required for rDNA silencing. In contrast to many other rDNA silencing mutants that have elevated levels of rDNA recombination, snf2Delta mutants have a significantly decreased level of rDNA recombination. Additional studies have demonstrated that Swi/Snf is also required for silencing of genes near telomeres while having no detectable effect on silencing of HML or HMR. PMID- 15340083 TI - Brap2 functions as a cytoplasmic retention protein for p21 during monocyte differentiation. AB - The cell cycle inhibitor p21 plays an important role in monocytic cell differentiation, during which it translocates from the nucleus to cytoplasm. This process involves the negative regulation of the p21 nuclear localization signal (NLS). Here, we sought to determine the relationship between the cytoplasmic translocation of p21 and another molecule, Brap2, a cytoplasmic protein which binds the NLS of BRCA1 and was recently reported to inactivate KSR in the Ras activating signal pathway under the name of IMP. We report that p21 and Brap2 directly interact, both in vitro and in vivo, in a manner requiring the NLS of p21 and the C-terminal portion of Brap2. When it is cotransfected with Brap2, p21 is expressed in the cytoplasm. Monocytic differentiation of the promyelomonocytic cell lines U937 and HL60 is associated with the upregulation of Brap2 expression concomitantly with the upregulation and cytoplasmic relocalization of p21. Our results underscore the role played by Brap2 in the process of cytoplasmic translocation of p21 during monocyte differentiation. PMID- 15340084 TI - Structural and functional organization of TRAP220, the TRAP/mediator subunit that is targeted by nuclear receptors. AB - The TRAP/Mediator complex serves as a coactivator for many transcriptional activators, including nuclear receptors such as the thyroid hormone receptor (TR) that targets the TRAP220 subunit. The critical but selective function of TRAP220 is evidenced by the embryonic lethal phenotype of Trap220(-)(/)(-) mice and by the observation that Trap220(-)(/)(-) fibroblasts (isolated before embryonic death) are impaired in specific nuclear receptor-dependent pathways. Here we have used a biochemical and genetic approach to understand the basis of specificity in TRAP220 function. We show that Trap220(-)(/)(-) cells possess a TRAP/Mediator complex that is relatively intact and compromised in its ability to support TR dependent, but not VP16-dependent, transcription in vitro. Transfection studies using TRAP220 mutants revealed that the N terminus of TRAP220 is necessary and sufficient for stable association with the TRAP/Mediator complex and, further, that TRAP220-dependent TR function in transfected cells requires both of the NR boxes that contain the LXXLL motif implicated in nuclear receptor binding. Similarly, an analysis of isolated TRAP/Mediator complexes with mutations in either or both of the two NR boxes confirmed a critical role for them in in vitro coactivator function. The implications of these observations are discussed in terms of our present understanding of coactivator function. PMID- 15340085 TI - Pak1 protein kinase regulates activation and nuclear localization of Snf1-Gal83 protein kinase. AB - Three kinases, Pak1, Tos3, and Elm1, activate Snf1 protein kinase in Saccharomyces cerevisiae. This cascade is conserved in mammals, where LKB1 activates AMP-activated protein kinase. We address the specificity of the activating kinases for the three forms of Snf1 protein kinase containing the beta subunit isoforms Gal83, Sip1, and Sip2. Pak1 is the most important kinase for activating Snf1-Gal83 in response to glucose limitation, but Elm1 also has a significant role; moreover, both Pak1 and Elm1 affect Snf1-Sip2. These findings exclude the possibility of a one-to-one correspondence between the activating kinases and the Snf1 complexes. We further identify a second, unexpected role for Pak1 in regulating Snf1-Gal83: the catalytic activity of Pak1 is required for the nuclear enrichment of Snf1-Gal83 in response to carbon stress. The nuclear enrichment of Snf1 fused to green fluorescent protein (GFP) depends on both Gal83 and Pak1 and is abolished by a mutation of the activation loop threonine; in contrast, the nuclear enrichment of Gal83-GFP occurs in a snf1Delta mutant and depends on Pak1 only when Snf1 is present. Snf1-Gal83 is the only form of the kinase that localizes to the nucleus. These findings, that Pak1 both activates Snf1-Gal83 and controls its nuclear localization, implicate Pak1 in regulating nuclear Snf1 protein kinase activity. PMID- 15340086 TI - Phosphorylation and alternative pre-mRNA splicing converge to regulate myocyte enhancer factor 2C activity. AB - Myocyte enhancer factor 2 (MEF2) transcription factors play pivotal roles in cardiac, muscle, and neuron gene expression. All products of MEF2 genes have a common amino-terminal DNA binding and dimerization domain, but the four vertebrate MEF2 gene transcripts are alternatively spliced among coding exons to produce splicing isoforms. In MEF2C alone, alternative splice acceptors in the last exon give forms that include or exclude a short domain that we designate gamma. We show that MEF2C is expressed exclusively as gamma- isoforms in heart tissue and predominantly as gamma- in other adult tissues and in differentiating myocytes. MEF2C gamma- isoforms are much more robust than gamma+ forms in activating MEF2-responsive reporters in transfected fibroblasts despite indistinguishable expression levels, and they better synergize with MyoD in promoting myogenic conversion. One-hybrid transcription assays using Gal4-MEF2C fusions give similar distinctions between gamma- and gamma+ isoforms in all cell types tested, including myocytes. Cis effects of gamma on MEF2C DNA binding, dimerization, protein stability, or response to CaM or p38 mitogen-activated protein kinase signaling are not apparent, and the isolated gamma domain represses transcription when fused to Gal4. One phosphoserine residue is present within the gamma domain according to tandem mass spectrometry, and mutation of this residue abolishes gamma-mediated transrepression. A similar activity is present in the constitutive gamma domain and serine phosphoacceptor of MEF2A. Our findings indicate that gamma functions autonomously as a phosphoserine-dependent transrepressor to downregulate transactivation function of MEF2 factors and that alternative splicing and serine phosphorylation converge to provide complex combinatorial control of MEF2C activity. PMID- 15340087 TI - Yeast Asc1p and mammalian RACK1 are functionally orthologous core 40S ribosomal proteins that repress gene expression. AB - Translation of mRNA into protein is a fundamental step in eukaryotic gene expression requiring the large (60S) and small (40S) ribosome subunits and associated proteins. By modern proteomic approaches, we previously identified a novel 40S-associated protein named Asc1p in budding yeast and RACK1 in mammals. The goals of this study were to establish Asc1p or RACK1 as a core conserved eukaryotic ribosomal protein and to determine the role of Asc1p or RACK1 in translational control. We provide biochemical, evolutionary, genetic, and functional evidence showing that Asc1p or RACK1 is indeed a conserved core component of the eukaryotic ribosome. We also show that purified Asc1p-deficient ribosomes have increased translational activity compared to that of wild-type yeast ribosomes. Further, we demonstrate that asc1Delta null strains have increased levels of specific proteins in vivo and that this molecular phenotype is complemented by either Asc1p or RACK1. Our data suggest that one of Asc1p's or RACK1's functions is to repress gene expression. PMID- 15340088 TI - Selective coactivator interactions in gene activation by SREBP-1a and -1c. AB - Requisite levels of intracellular cholesterol and fatty acids are maintained in part by the sterol regulatory element binding proteins (SREBPs). Three major SREBP isoforms exist; SREBP-1a and SREBP-1c are expressed from overlapping mRNAs, whereas SREBP-2 is encoded by a separate gene. The active forms of SREBP-1a and SREBP-1c differ only at their extreme N termini; SREBP-1c lacks 28 aa present in SREBP-1a and instead contains 4 unique aa of its own. While the SREBP-1a and -1c isoforms differentially activate transcription, the molecular basis of this difference is unknown. Here we define the differences between these proteins that confer the enhanced activity of SREBP-1a and demonstrate that this enhancement is a direct result of its avid binding to the coactivator CREB binding protein (CBP) and the mammalian mediator complex. While previous work determined that the C/H1 zinc finger and KIX domains of CBP bind to SREBP-1a, we provide evidence that the interaction with C/H1 is important for gene activation. We further show that the association between the activation domain of SREBP-1 and mediator is through aa 500 to 824 of DRIP150. Finally, we demonstrate the recruitment of mediator to an SREBP-responsive promoter in a sterol-dependent manner. PMID- 15340089 TI - Early intermediates of mariner transposition: catalysis without synapsis of the transposon ends suggests a novel architecture of the synaptic complex. AB - The mariner family is probably the most widely distributed family of transposons in nature. Although these transposons are related to the well-studied bacterial insertion elements, there is evidence for major differences in their reaction mechanisms. We report the identification and characterization of complexes that contain the Himar1 transposase bound to a single transposon end. Titrations and mixing experiments with the native transposase and transposase fusions suggested that they contain different numbers of transposase monomers. However, the DNA protection footprints of the two most abundant single-end complexes are identical. This indicates that some transposase monomers may be bound to the transposon end solely by protein-protein interactions. This would mean that the Himar1 transposase can dimerize independently of the second transposon end and that the architecture of the synaptic complex has more in common with V(D)J recombination than with bacterial insertion elements. Like V(D)J recombination and in contrast to the case for bacterial elements, Himar1 catalysis does not appear to depend on synapsis of the transposon ends, and the single-end complexes are active for nicking and probably for cleavage. We discuss the role of this single-end activity in generating the mutations that inactivate the vast majority of mariner elements in eukaryotes. PMID- 15340091 TI - Low sodium haemodialysis reduces interdialytic fluid consumption but paradoxically increases post-dialysis thirst. AB - BACKGROUND: Interdialytic weight gain (IDWG) can be reduced by lowering the dialysate sodium concentration ([Na]) in haemodialysis patients. It has been assumed that this is because thirst is reduced, although this has been difficult to prove. We compared thirst patterns in stable haemodialysis patients with high and low IDWG using a novel technique and compared the effect of low sodium dialysis (LSD) with normal sodium dialysis (NSD). METHODS: Eight patients with initial high IDWG and seven with low IDWG completed hourly visual analogue ratings of thirst using a modified palmtop computer during the dialysis day and the interdialytic day. The dialysate [Na] was progressively reduced by up to 5 mmol/l over five treatments. Dialysis continued at the lowest attained [Na] for 2 weeks and the measurements were repeated. The dialysate [Na] then returned to baseline and the process was repeated. RESULTS: Baseline interdialytic day mean thirst was higher than the dialysis day mean for the high IDWG group (49.9+/-14.0 vs 36.2+/-16.6) and higher than the low weight gain group (49.9+/-14.0 vs 34.1+/ 14.6). This trend persisted on LSD, but there was a pronounced increase in post dialysis thirst scores for both groups (high IDWG: 46+/-13 vs 30+/-21; low IDWG: 48+/-24 vs 33+/-18). The high IDWG group demonstrated lower IDWG during LSD than NSD (2.23+/-0.98 vs 2.86+/-0.38 kg; P<0.05). CONCLUSIONS: Our results indicate that patients with high IDWG experience more intense feelings of thirst on the interdialytic day. LSD reduces their IDWG, but paradoxically increases thirst in the immediate post-dialysis period. PMID- 15340090 TI - Role for Nhp6, Gcn5, and the Swi/Snf complex in stimulating formation of the TATA binding protein-TFIIA-DNA complex. AB - The TATA-binding protein (TBP), TFIIA, and TFIIB interact with promoter DNA to form a complex required for transcriptional initiation, and many transcriptional regulators function by either stimulating or inhibiting formation of this complex. We have recently identified TBP mutants that are viable in wild-type cells but lethal in the absence of the Nhp6 architectural transcription factor. Here we show that many of these TBP mutants were also lethal in strains with disruptions of either GCN5, encoding the histone acetyltransferase in the SAGA complex, or SWI2, encoding the catalytic subunit of the Swi/Snf chromatin remodeling complex. These synthetic lethalities could be suppressed by overexpression of TOA1 and TOA2, the genes encoding TFIIA. We also used TFIIA mutants that eliminated in vitro interactions with TBP. These viable TFIIA mutants were lethal in strains lacking Gcn5, Swi2, or Nhp6. These lethalities could be suppressed by overexpression of TBP or Nhp6, suggesting that these coactivators stimulate formation of the TBP-TFIIA-DNA complex. In vitro studies have previously shown that TBP binds very poorly to a TATA sequence within a nucleosome but that Swi/Snf stimulates binding of TBP and TFIIA. In vitro binding experiments presented here show that histone acetylation facilitates TBP binding to a nucleosomal binding site and that Nhp6 stimulates formation of a TBP-TFIIA DNA complex. Consistent with the idea that Nhp6, Gcn5, and Swi/Snf have overlapping functions in vivo, nhp6a nhp6b gcn5 mutants had a severe growth defect, and mutations in both nhp6a nhp6b swi2 and gcn5 swi2 strains were lethal. PMID- 15340092 TI - Acute effect of haemodialysis on arterial stiffness: membrane bioincompatibility? AB - BACKGROUND: Repetitive endothelial damage from dialysis membrane incompatibility is a probable cause of accelerated atherosclerosis in haemodialysis patients. Consequently pulse wave velocity (PWV), a measure of arterial stiffness, was utilized as a surrogate marker of vascular dysfunction during dialysis with two commonly used synthetic dialysers. METHODS: PWV was monitored before, during and after haemodialysis using both polysulphone and polyamide membranes. PWV, an arterial stiffness measure, was calculated from the carotid to the femoral (C-F) and also to the radial (C-R) artery. In a further group, PWV was monitored while polysulphone and polyamide membranes were perfused with blood without dialysate. RESULTS: Mean aortic (C-F) PWV was lower during dialysis with the polyamide membrane, being 14 and 16% less following 75 and 135 min of dialysis (P<0.05) in 24 patients. Because intradialytic intravascular volume changes alter PWV, a subgroup analysis in 11 patients where dialysis fluid removal during both periods was minimal (<1 kg) was performed, and a persistent and significant increase in aortic PWV was detected with the polysulphone kidney being maximal (40%) at 75 min (P<0.01). This increase was negatively correlated with pre-dialysis PWV (P<0.01). In contrast, the polyamide dialyser did not change PWV. An increase in C-R PWV was also noted with the polysulphone membrane (P<0.05). In the nine patients where membranes were perfused with blood without dialysate, aortic PWV was again significantly increased by the polysulphone (P<0.01), but not the polyamide dialyser. CONCLUSIONS: Haemodialysis with polysulphone but not polyamide membranes acutely alters aortic 'stiffness', an effect postulated to be due to membrane bioincompatibility. However, factors including age, time on dialysis and underlying vascular disease, were also found to impact on these acute dialysis-induced changes to vascular function. Since these acute changes disappear post-dialysis, their long-term consequences are uncertain. PMID- 15340093 TI - Injection of recombinant FcalphaRI/CD89 in mice does not induce mesangial IgA deposition. AB - BACKGROUND: Earlier studies have suggested that complexes of the human IgA receptor FcalphaRI/CD89 with mouse IgA are pathogenic upon deposition in the renal mesangium. Transgenic mice expressing FcalphaRI/CD89 on macrophages/monocytes developed massive mesangial IgA deposition and a clinical picture of IgA nephropathy (IgAN). Based on these findings, the purpose of this study was to design an experimental model of IgAN by injection of human CD89 in mice. The interaction of mouse IgA with CD89 was investigated further. METHODS: Recombinant human soluble CD89 and a chimeric CD89-Fc protein were generated, produced, purified and injected in mice. Renal cryosections were stained for IgA and CD89. The interaction of mouse IgA with CD89 was analysed by fluorescence activated cell sorting (FACS) analysis, enzyme-linked immunosorbent assay (ELISA) and plasmon resonance technology. RESULTS: Injection of recombinant human CD89 did not result in significant IgA or CD89 deposition in the renal mesangium. However, CD89 staining in the liver was found to be positive. CD89 was rapidly cleared from circulation without signs of complex formation with IgA. FACS analysis, ELISA and plasmon resonance techniques all revealed a dose-dependent binding of human IgA to recombinant CD89, while no detectable binding was seen of mouse IgA, either of serum IgA or of different monoclonal mouse IgA preparations. CONCLUSIONS: An experimental model for IgAN in mice could not be obtained by injection of recombinant CD89. This is compatible with our in vitro biochemical data showing a lack of binding between recombinant human CD89 and mouse IgA. PMID- 15340094 TI - Tesio-Caths provide effective and safe long-term vascular access. AB - BACKGROUND: Vascular access is judged on its ability to provide good dialysis adequacy, its durability and complication rates. Formation of a functional arteriovenous fistula is desirable but difficult to achieve in a significant proportion of patients. We report the large-scale use of Tesio-Caths, a twin-line single-lumen central venous catheter, to maximize dialysis adequacy where formation of an arteriovenous fistula was not possible. METHODS: All patients who had Tesio-Caths inserted between 1 January 1999 and 1 October 2002 were studied. RESULTS: Six hundred and twenty-three Tesio-Caths were inserted from 1 January 1999 to 1 October 2002 in 435 patients, generating 7464 patient months of follow up. Five hundred and ninety-four out of 623 (95.3%) Tesio-Caths were immediately functional. Mean dialysis adequacy measured by single-pool Kt/V was 1.5+/-0.3 for all Tesio-Caths for the entire period of study, with 68% of Tesio-Caths delivering a Kt/V >1.4. Cumulative functional Tesio-Cath survival to final failure was 77.8 and 44% at 1 and 3 years, respectively. Cumulative patient survival was 84.7, 71.4 and 63% at 1, 2 and 3 years, respectively. Access-related infection accounted for 0.28 admissions/1000 catheter days, and the death rate from access-related sepsis was 9.6 deaths/1000 patient years at risk. The admission rate for access dysfunction was 0.33/1000 patient years at risk. CONCLUSION: Tesio-Caths provide good dialysis adequacy for patients in whom an arteriovenous fistula cannot be formed. Patient and functional access survival for this group was comparable with current European data irrespective of vascular access type. Complication rates were acceptably low. PMID- 15340095 TI - Intensive treatment with statins and the progression of cardiovascular diseases: the beginning of a new era? PMID- 15340096 TI - Interleukin-18 is a strong predictor of hospitalization in haemodialysis patients. AB - BACKGROUND: Morbidity and mortality rates are high among patients with end-stage renal disease (ESRD), and recent evidence suggests that this may be linked to inflammation. Current research has also demonstrated the crucial involvement of interleukin-18 (IL-18) in inflammation. In agreement, the activity of IL-18 has been markedly up-regulated in ESRD patients. However, it has not been established whether elevated plasma IL-18 predicts outcome in haemodialysis (HD) patients. METHODS: To determine whether plasma IL-18 predicts overall hospitalization, we studied 184 ESRD patients (62% males, 58.5+/-1.0 years of age) undergoing maintenance HD treatment. The patients were followed for 12 months and were stratified by the tertiles of plasma IL-18 levels. Classic factors, such as age, body mass index, duration of HD, nutritional and inflammatory parameters, co morbidity, dialysis adequacy, and lipid status were entered into a Cox regression model to predict hospitalization. The Kaplan-Meier method was used to analyse the cumulative proportion of hospitalization-free events. RESULTS: Significantly different hospitalization days and frequencies (P<0.05) were observed when patients were divided according to tertiles of plasma IL-18 levels. Patients were stratified according to IL-18 tertiles and analysed separately according to the hospitalization-free period. In the Kaplan-Meier model, the upper tertile of IL 18 had the highest probability of a hospitalization event during the entire follow-up period (P log rank = 0.027). In the Cox proportional hazard model, the relative risk for first hospital admission for each increase in Ln IL-18 (pg/ml) concentration was associated with a 1.709 (95% CI, 1.114 to 2.620; P = 0.014) increase in the risk for future hospitalization events. CONCLUSIONS: The present study demonstrated a strong predictive value of elevated IL-18 levels for poor outcome in HD patients. PMID- 15340097 TI - Effect of lactate and bicarbonate on human peritoneal mesothelial cells, fibroblasts and vascular endothelial cells, and the role of basic fibroblast growth factor. AB - BACKGROUND: In patients on long-term continuous ambulatory peritoneal dialysis (CAPD), peritoneal dysfunction may occur due to loss of peritoneal mesothelial cells, peritoneal fibrosis and neovascularization. Lactate, long used as a buffer in peritoneal dialysates, has been substituted by bicarbonate in recent years. However, their effects on the peritoneum of CAPD patients are unknown. This study investigated the influence of lactate and bicarbonate on peritoneal dysfunction in CAPD patients. METHODS: The mitochondrial activity of human peritoneal mesothelial cells (HPMCs) and their expression of basic fibroblast growth factor (bFGF) were studied after culture under various conditions. We also assessed the mitochondrial-activating effect of the supernatant of those cultures on human peritoneal fibroblasts (HPFBs) and human umbilical vein endothelial cells (HUVECs) and the effect of recombinant human bFGF on the mitochondrial activity of HPFBs and HUVECs. We used the WST-1 assay to determine mitochondrial activity in HPMC. RESULTS: At pH 7.4, the mitochondrial activity of HPMCs was lowest in a medium containing 40 mM (Lac), intermediate in a lactate (15 mM) plus bicarbonate (25 mM) medium (Lac/Bic), and highest in a 40 mM bicarbonate medium (Bic). In culture supernatant, the increase of bFGF was: Lac > Lac/Bic > Bic. Mitochondrial activation of HPFBs and HUVECs was stimulated by HPMC culture supernatants in the following decreasing order: Lac > Lac/Bic > Bic. The effects of these supernatants were suppressed by a bFGF-neutralizing antibody, while recombinant bFGF caused concentration-dependent mitochondrial activation in HPFBs and HUVECs. CONCLUSIONS: The role of bFGF in peritoneal fibrosis and neovascularization may be important. A bicarbonate-containing medium is better than a lactate-containing medium for preserving cell viability in HPMCs and preventing bFGF expression by these cells. PMID- 15340098 TI - Acute interstitial nephritis: clinical features and response to corticosteroid therapy. AB - BACKGROUND: Acute interstitial nephritis (AIN) is a recognized cause of reversible acute renal failure characterized by the presence of an interstitial inflammatory cell infiltrate. METHODS: In order to evaluate the clinical characteristics and management of this disorder, we performed a retrospective study of all cases of AIN found by reviewing 2598 native renal biopsies received at our institution over a 12 year period. Presenting clinical, laboratory and histological features were identified, as was clinical outcome with specific regard to corticosteroid therapy response. RESULTS: AIN was found in 2.6% of native biopsies, and 10.3% of all biopsies performed in the setting of acute renal failure during the period analysed (n = 60). The incidence of AIN increased progressively over the period observed from 1 to 4% per annum. AIN was drug related in 92% of cases and appeared to be idiopathic in the remainder. The presenting symptoms included oliguria (51%), arthralgia (45%), fever (30%), rash (21%) and loin pain (21%). Median serum creatinine at presentation was 670 micromol/l [interquartile range (IQR) 431-1031] and 58% of cases required acute renal replacement therapy. Corticosteroid therapy was administered in 60% of cases. Serum creatinine at baseline was similar in the corticosteroid-treated and conservatively managed groups; 700 micromol/l (IQR 449-1031) vs 545 micromol/l (IQR 339-1110) P = 0.4. In this, the largest retrospective series to date, we did not detect a statistically significant difference in outcome, as determined by serum creatinine, between those patients who received corticosteroid therapy and those who did not, at 1, 6 and 12 months following presentation. CONCLUSION: The results of this study do not support the routine administration of corticosteroid therapy in the management of AIN. PMID- 15340099 TI - Addressing the unmet medical need for safe and effective weight loss therapies. AB - Obesity is a significant healthcare problem worldwide and increases the risk of developing debilitating diseases including type 2 diabetes, cardiovascular disease, and cancer. Although the health benefits of weight reduction are well recognized, weight loss by diet and exercise fail in most patients, and the current marketed drugs have had limited success. It is clear that there is a significant unmet medical need for safe and effective weight-reducing agents. In this review, the current status of potential weight loss approaches that are in development by the pharmaceutical and biotechnology industry are discussed. This should lead to novel treatments that can be used long-term to effectively treat this serious metabolic disorder. PMID- 15340100 TI - Current and investigational antiobesity agents and obesity therapeutic treatment targets. AB - Public health efforts and current antiobesity agents have not controlled the increasing epidemic of obesity. Investigational antiobesity agents consist of 1) central nervous system agents that affect neurotransmitters or neural ion channels, including antidepressants (bupropion), selective serotonin 2c receptor agonists, antiseizure agents (topiramate, zonisamide), some dopamine antagonists, and cannabinoid-1 receptor antagonists (rimonabant); 2) leptin/insulin/central nervous system pathway agents, including leptin analogues, leptin transport and/or leptin receptor promoters, ciliary neurotrophic factor (Axokine), neuropeptide Y and agouti-related peptide antagonists, proopiomelanocortin and cocaine and amphetamine regulated transcript promoters, alpha-melanocyte stimulating hormone analogues, melanocortin-4 receptor agonists, and agents that affect insulin metabolism/activity, which include protein-tyrosine phosphatase-1B inhibitors, peroxisome proliferator activated receptor-gamma receptor antagonists, short-acting bromocriptine (ergoset), somatostatin agonists (octreotide), and adiponectin; 3) gastrointestinal-neural pathway agents, including those that increase cholecystokinin activity, increase glucagon-like peptide-1 activity (extendin 4, liraglutide, dipeptidyl peptidase IV inhibitors), and increase protein YY3-36 activity and those that decrease ghrelin activity, as well as amylin analogues (pramlintide); 4) agents that may increase resting metabolic rate ("selective" beta-3 stimulators/agonist, uncoupling protein homologues, and thyroid receptor agonists); and 5) other more diverse agents, including melanin concentrating hormone antagonists, phytostanol analogues, functional oils, P57, amylase inhibitors, growth hormone fragments, synthetic analogues of dehydroepiandrosterone sulfate, antagonists of adipocyte 11B hydroxysteroid dehydrogenase type 1 activity, corticotropin-releasing hormone agonists, inhibitors of fatty acid synthesis, carboxypeptidase inhibitors, indanones/indanols, aminosterols, and other gastrointestinal lipase inhibitors (ATL962). Finally, an emerging concept is that the development of antiobesity agents must not only reduce fat mass (adiposity) but must also correct fat dysfunction (adiposopathy). PMID- 15340101 TI - Association of cholecystokinin 1 receptor and beta3-adrenergic receptor polymorphisms with midlife weight gain. AB - We investigated the relationship of polymorphisms in the cholecystokinin 1 receptor [CCK1R; G to T (n-128), A to G (n-81)] and the beta3-adrenergic receptor (beta3-AR; Trp64Arg) with midlife weight gain. The participants were 1012 Japanese men and women (40 to 59 years of age). Their weight at 18 years old was obtained from a questionnaire. Weight change was defined as the current weight minus the weight at 18 years old. Subjects were grouped into four categories by these genotypes: W/W=noncarriers, W/H=Arg64 carriers of the beta3-AR, H/W=T (n 128) or G (n-81) carriers of the CCK1R, H/H=T (n-128) or G (n-81) and Arg64 carriers. In men, the interaction between the CCK1R and beta3-AR polymorphisms was significant (two-way ANOVA, p < 0.05), but neither the CCK1R nor the beta3-AR was individually associated with weight gain. The H/H group showed a higher possibility of weight gain of 10 kg or more compared with the W/W group in men. The odds ratio for weight gain (> or =10 kg) of H/H was 2.54 (95% confidence interval: 1.50 to 4.30) compared with W/W. In women, neither main effect nor interaction was significant. These results suggest that the combination of CCK1R and the beta3-AR polymorphisms is a contributing factor for midlife weight gain in men. PMID- 15340102 TI - A survey of genes differentially expressed in subcutaneous and visceral adipose tissue in men. AB - Adipose tissue located within the abdominal cavity has been suggested to be functionally and metabolically distinct from that of the subcutaneous compartment. These differences could play a role in obesity-related complications. The aim of this study was to compare gene expression profiles of subcutaneous and visceral adipose tissues of 10 nondiabetic, normolipidemic obese men. Affymetrix human U133A arrays (10 arrays for subcutaneous fat samples and 10 arrays for visceral fat samples) were used. Differential gene expression was confirmed by real-time polymerase chain reaction in a subset of genes. A total of 5894 transcripts were detected in both depots in all 10 subjects, and 409 transcripts representing 347 encoded genes were differentially expressed. Of these, 131 genes were expressed at higher levels in subcutaneous adipose tissue, and 216 were expressed more abundantly in visceral fat. Differentially expressed profiles included genes of the Wnt signaling pathway, as well as CEPBA and HOX genes. In addition, genes involved in lipolytic stimuli and cytokine secretion were differentially expressed. The identification of a consistent and rather uniform pattern of differentially expressed genes between the two fat depots using multiple array replicates (10 arrays per fat compartment) generated new perspectives for future research on regional differences in adipose tissue biology. PMID- 15340103 TI - Leptin and nitric oxide production in normotensive and hypertensive men. AB - OBJECTIVE: Recent findings have shown that leptin, the product of the obesity gene, may actively participate in the regulation of blood pressure and other cardiovascular functions through the nitric oxide (NO)-dependent mechanism. RESEARCH METHODS AND PROCEDURES: In this study, to test the hypothesis that leptin regulation of NO metabolism is impaired in hypertension, we examined the possible relationship between circulating leptin and plasma NO metabolite level in normotensive (NT) and hypertensive (HT) men. RESULTS: There were significant correlations between circulating leptin and BMI in both the NT and HT groups (NT: r = 0.64, n = 26, p < 0.01; HT: r = 0.59, n = 22, p < 0.01). The concentration of circulating leptin was similar between the NT and HT men, although the plasma NO metabolite level (nitrite and nitrate) was significantly reduced in the HT men compared with the NT men (NT: 51.0 +/- 4.9 microM, n = 26; HT: 37.1 +/- 2.5 microM, n = 22, p < 0.05). The circulating leptin was significantly correlated with the plasma NO metabolite level in the overall analysis of the NT and HT men (r = 0.35, n = 48, p < 0.05). When the analysis of the correlation for the NT and HT men was performed separately, there was a significant correlation between circulating leptin and plasma NO metabolites in the NT men (r = 0.45, n = 26, p < 0.05) but not in the HT men (r = 0.15, n = 22). The results of this study are consistent with the hypothesis that leptin-related metabolism of NO might be altered in HT men. PMID- 15340104 TI - Rate of fat gain is faster in girls undergoing early adiposity rebound. AB - OBJECTIVE: To determine the changes in body composition (fat and lean mass) occurring in children during adiposity rebound (AR). RESEARCH METHODS AND PROCEDURES: Thirty-nine girls, 3 to 6 years of age at baseline, underwent yearly DXA scans for 2 years. An additional DXA scan was obtained 4 to 5 years after baseline. Age at AR was determined by modeling, and the velocity of change in height, weight, fat mass, and lean mass was estimated for each child using random coefficient models. Girls with an AR <5 years of age were classified as having an early AR, and those having an AR > or =5 years were classified as late AR. RESULTS: Although body composition was similar at age 5, by age 9, girls with an early AR were significantly taller (3.5% more) and heavier (14.4%), with greater fat mass (50%) and percentage body fat (27%) than girls with a later AR. In addition, more girls were overweight according to BMI (18% vs. 6%) or percentage body fat (29% vs. 11%) at this time, despite no differences at baseline. Annual velocity of fat mass gain was over 2-fold higher in early compared with late rebounders (17.1% vs. 6.5%, p < 0.0001), with no difference in lean mass velocity (13.1% vs. 12.5%, p = 0.116). DISCUSSION: Differences in BMI during AR were caused specifically by alterations in body fat and not by alterations in lean mass or height. Children undergoing early AR gained fat at a faster rate than children who rebounded at a later age. PMID- 15340105 TI - Adipogenic and antiapoptotic protein levels in human adipose stromal cells after weight loss. AB - OBJECTIVE: Obesity is a major risk factor for type 2 diabetes and cardiovascular disease. However, current strategies to achieve sustained weight loss are often unsuccessful. Fat reaccumulation might be favored by enhanced adipose cell differentiation or survival in the postreduced state. RESEARCH METHODS AND PROCEDURES: We measured adipogenic and apoptotic protein expression in subcutaneous abdominal adipose stromal-vascular cells from 10 obese patients (7 women and 3 men) that were obtained before and after a 16% weight loss in a medically supervised weight loss program. RESULTS: After weight loss, protein expression was 2.4-fold higher (p < 0.005) for p42 C/CAAT enhancer binding protein alpha, but there was no change for peroxisome proliferator-activated receptor gamma1; both of these are adipogenic regulators. For neuronal apoptosis inhibitory protein, a protein associated with adipose cell apoptotic resistance, there was a rise of 1.7-fold (p < 0.02). DISCUSSION: Alterations in C/CAAT enhancer binding protein alpha and neuronal apoptosis inhibitory protein expression occurred in human adipose stromal-vascular cells after weight loss in a pilot study of 10 patients. It will be important for future studies to directly examine whether the adipogenic and antiapoptotic capacity of these cells is changed after weight loss. PMID- 15340106 TI - Intestinal electric stimulation decreases fat absorption in rats: therapeutic potential for obesity. AB - OBJECTIVE: Effective treatment of obesity is based on the restriction of food intake or reduction of absorption or both. The aim of this study was to study whether intestinal electric stimulation (IES) would reduce fat absorption and, thus, would be a potential therapy for obesity. RESEARCH METHODS AND PROCEDURES: Forty rats implanted with serosal electrodes and two jejunal cannulas were divided into 4 groups of 10 each: control (no stimulation), IES with long pulses, IES with trains of short pulses, and IES with trains of short pulses plus treatment with lidocaine. Jejunal transit and fat absorption of a 20-cm jejunal segment (between two cannulas) were investigated during a 45-minute period with or without IES. RESULTS: It was found that both methods of IES accelerated intestinal transit measured by recovery of phenol red and increased the percentage of triglycerides recovered from the distal cannula in comparison with the control group. IES with trains of short pulses was more effective than IES with long pulses in accelerating jejunal transit and reducing fat absorption. Neither of the two IES methods altered the output of fatty acids from the distal cannula. The effects of IES with trains of short pulses on the transit and fat absorption were partially abolished with the treatment of lidocaine. DISCUSSION: It was concluded that IES accelerates intestinal transit and reduces fat absorption, suggesting a therapeutic potential for obesity. IES with trains of short pulses is more effective than IES with long pulses, and its effects are partially mediated by enteric nerves, jejunum. PMID- 15340107 TI - The yellow agouti mutation alters some but not all responses to diet and exercise. AB - OBJECTIVE: Effects of ectopic expression of the agouti signaling protein were studied on responses to diet restriction and exercise in C57BL/6J (B6) mice and obese B6 mice congenic for the yellow agouti mutation [B6.Cg-Ay (Ay)]. RESEARCH METHODS AND PROCEDURES: Adult male Ay mice were either kept sedentary or exercised on a running wheel and fed ad libitum or diet restricted until weight matched to ad libitum-fed B6 control mice. Body composition, plasma lipids, leptin, and adiponectin were measured. mRNA levels for leptin, adiponectin, lipoprotein lipase, and pyruvate dehydrogenase kinase 4 were measured in a visceral (epididymal) and a subcutaneous (femoral) fat depot by real-time polymerase chain reaction. RESULTS: Correlations among traits exhibited one of three patterns: similar lines for B6 and Ay mice, different slopes for B6 and Ay mice, and/or different intercepts for B6 and Ay mice. Correlations involving plasma leptin, mesenteric and epididymal adipose weights, or low-density lipoprotein-cholesterol were most likely to have different slopes and/or intercepts in B6 and Ay mice. mRNA levels for leptin, Acrp30, pyruvate dehydrogenase kinase 4, and lipoprotein lipase in epididymal adipose tissue were not correlated with corresponding levels in femoral adipose tissue. DISCUSSION: The agouti protein interferes with leptin signaling at melanocortin receptors in the hypothalamus of Ay mice. Our results are consistent with the hypothesis that the melanocortin portion of the leptin-signaling pathway mediates effects primarily on certain fat depots and on some, but not all, components of cholesterol homeostasis. PMID- 15340108 TI - Short-term effects of weight loss on the cardiovascular risk factors in morbidly obese patients. AB - OBJECTIVE: To analyze the short-term effects of weight loss on the cardiovascular risk factors in morbidly obese patients. RESEARCH METHODS AND PROCEDURES: Five metabolic cardiovascular risk factors (blood glucose, blood pressure, total cholesterol, high-density lipoprotein (HDL)-cholesterol, and triglycerides) were determined before and 15.3 +/- 2.1 months after laparoscopic gastric banding in 650 morbidly obese patients. Global cardiovascular risk was calculated according to the Prospective Cardiovascular Munster (PROCAM) scoring system. RESULTS: Mean weight loss was 22.7 +/- 20.4 kg. Normalization of the metabolic alteration was observed in 67.3% of patients with diabetes, 38.3% of patients with hypercholesterolemia, 72.5% of patients with low HDL-cholesterol, 72.3% of patients with hypertriglyceridemia, and 46.7% of patients with hypertension. PROCAM score fell from 31.4 +/- 11.6 to 28.0 +/- 12.0 points (p < 0.001). The modifications of total cholesterol and blood pressure were unrelated to percentage weight loss. Percentage weight loss was significantly related to the reductions of fasting blood glucose, triglyceride level, and the PROCAM score and to the increase of HDL-cholesterol concentrations observed after surgery. However, the strength of these four relationships was generally low. The variations of HDL-cholesterol concentrations and blood pressure levels were more influenced by actual energy balance than by the extent of weight loss. DISCUSSION: Weight loss observed in the first 12 to 18 months after gastric banding was associated with a significant improvement of single cardiovascular risk factors and global risk. On the other hand, the extent of weight loss was poorly related to the magnitude of improvement in cardiovascular risk. PMID- 15340109 TI - Diet-induced changes in stearoyl-CoA desaturase 1 expression in obesity-prone and -resistant mice. AB - OBJECTIVE: To investigate stearoyl-coenzyme A desaturase (SCD) 1 expression in obesity-prone C57BL/6 mice and in obesity-resistant FVB mice to explore the relationship of SCD1 expression and susceptibility to diet-induced obesity. RESEARCH METHODS AND PROCEDURES: Nine-week-old C57BL/6 and FVB mice were fed either a high- or low-fat diet for 8 weeks. Body weight and body composition were measured before and at weeks 4 and 8 of the study. Energy expenditure was measured at weeks 1 and 5 of the study. Hepatic SCD1 mRNA was measured at 72 hours and at the end of study. Plasma leptin and insulin concentrations were measured at the end of study. RESULTS: When C57BL/6 mice were switched to a calorie-dense high-fat diet, animals gained significantly more body weight than those maintained on a low-calorie density diet primarily due to increased fat mass accretion. Fat mass continued to accrue throughout 8 weeks of study. Increased calorie intake did not account for all weight gain. On the high-fat diet, C57BL/6 mice decreased their energy expenditure when compared with mice fed a low-fat diet. In response to 8 weeks of a high-fat diet, SCD1 gene expression in liver increased >2-fold. In contrast, feeding a high-fat diet did not change body weight, energy expenditure, or SCD1 expression in FVB mice. DISCUSSION: Our study showed that a high-fat hypercaloric diet increased body adiposity first by producing hyperphagia and then by decreasing energy expenditure of mice susceptible to diet-induced obesity. Consumption of a high-fat diet in species predisposed to obesity selectively increased SCD1 gene expression in liver. PMID- 15340110 TI - Promoting more modest weight losses: a pilot study. AB - OBJECTIVE: This pilot study assessed the short- and long-term effects of a modified cognitive behavioral treatment designed to facilitate obese patients' acceptance of a 5% to 10% reduction in initial weight. RESEARCH METHODS AND PROCEDURES: Participants were 17 women with a mean age of 46.5 +/- 9.7 years and BMI of 34.7 +/- 2.9 kg/m2. They participated in a 40-week program that included four phases. The first discussed the benefits of modest weight losses and the potential adverse effects of unrealistic expectations. Phase II provided instruction in traditional cognitive behavioral methods of weight control Phase III focused on methods to improve body image and self-esteem. Phase IV addressed skills for weight maintenance. Changes in weight, self-esteem, body image, and quality of life were assessed at the end of treatment and 1 year later (week 92). RESULTS: At week 40, participants lost an average of 5.7 +/- 5.3% of initial weight, which was associated with significant improvements in body image, self esteem, and quality of life. Improvements in psychosocial status were maintained at week 92, although mean weight loss at this time had declined to 2.9 +/- 5.6% of initial weight. Increased satisfaction with body weight at week 40 was associated with significantly better maintenance of weight loss at follow-up (r = -0.70; p = 0.02). DISCUSSION: Having participants seek only modest initial weight losses does not appear to facilitate weight maintenance. However, increasing patients' satisfaction with their body weight at the end of treatment may help improve weight maintenance. More research is needed on the relation between satisfaction with initial weight loss and long-term success. PMID- 15340111 TI - Altered kidney CYP2C and cyclooxygenase-2 levels are associated with obesity related albuminuria. AB - OBJECTIVE: To determine cytochrome P450 (CYP450) and cyclooxygenase (COX) expression and metabolite regulation and renal damage in the early stages of obesity-related hypertension and diabetes. RESEARCH METHODS AND PROCEDURES: Obese and lean Zucker rats at 10 to 12 weeks of age were studied. Blood pressure was measured in the conscious state using radiotelemetry. Blood glucose levels and body weight were measured periodically. Protein expression of CYP450 and COX enzymes in the kidney cortex, renal microvessels, and glomeruli was studied. The levels of CYP450 and COX metabolites in urine were measured, and urinary albumin excretion, an indicator of kidney damage, was measured. RESULTS: Body weight and blood glucose averaged 432 +/- 20 grams and 105 +/- 5 mg/dl, respectively, in obese Zucker rats as compared with 320 +/- 8 grams and 91 +/- 5 mg/dl, respectively, in age-matched 10- to 12-week-old lean Zucker rats. Renal microvascular CYP4A and COX-2 protein levels were increased 2.3- and 17.0-fold, respectively, in obese Zucker rats. The protein expression of CYP2C11 and CYP2C23 was decreased 2.0-fold in renal microvessels isolated from obese Zucker rats when compared with lean Zucker rats. The urinary excretion rate of thromboxane B(2) was increased significantly in obese Zucker as compared with lean Zucker rats (22.0 +/- 1.8 vs. 13.4 +/- 1.0 ng/d). Urinary albumin excretion, an index of kidney damage, was increased in the obese Zucker rat at this early age. DISCUSSION: These results suggest that increased CYP4A and COX-2 protein levels and decreased CYP2C11 and CYP2C23 protein levels occur in association with microalbuminuria during the onset of obesity-related hypertension and type 2 diabetes. PMID- 15340112 TI - The novel antiobesic HMR1426 reduces food intake without affecting energy expenditure in rats. AB - OBJECTIVE: To determine the effect of acute and chronic administration of a new food intake-reducing compound (HMR1426) with novel mode of action (retardation of gastric emptying) on body weight development, food intake, and energy metabolism in rats. RESEARCH METHODS AND PROCEDURES: Adult male Shoe-Wistar rats were implanted with transponders allowing registration of body temperature (Tb) and locomotor activity. HMR1426 (10 or 50 mg/kg) was given orally, and acute (8 hours) and chronic (15 days) effects were measured on food intake, Tb, activity, total energy expenditure (indirect calorimetry), and epididymal adipose tissue mass. The effect of chronic treatment was compared with the effect of sibutramine (10 mg/kg). RESULTS: HMR1426 (50 mg/kg) caused an acute and chronic decrease of food intake. There was no effect on the level and daily pattern of total energy expenditure, Tb, and locomotor activity. Respiratory quotient was acutely decreased by HMR1426 due to reduced food intake. Chronic treatment with HMR1426 decreased weight gain by 31% and epididymal white fat by 24%. Sibutramine caused a respective reduction of 48% and 35%. Energy efficiency was not affected by HMR1426 in contrast to sibutramine, which reduced energy efficiency and transiently increased activity. DISCUSSION: HMR1426 showed an anorectic potential in rats and decreased body weight and fat mass. This was achieved solely by reducing food intake without influencing overall energy expenditure or behavior suggesting a peripheral mode of action. Thus, HMR1426 can be considered a potential new drug for obesity treatment. PMID- 15340113 TI - Prevalence of overweight among inner city Hispanic-American children and adolescents. AB - OBJECTIVE: National surveys have pointed to a particularly high risk of pediatric overweight among U.S. Hispanics. However, the data have been primarily from the Mexican-American community. We studied the prevalence of overweight and clinical comorbidities in children and youth of predominantly El Salvadoran ancestry. RESEARCH METHODS AND PROCEDURES: A sample of 309 Hispanic youth, 6-18 years was surveyed from two inner city Washington, DC, clinics. BMI; triceps skinfold (TSF) and subscapular skinfold thickness (SSSF); bioelectrical impedance analysis (BIA); and blood pressure measures were obtained, along with information regarding physical activity, sedentary behavior, dietary history, family, and personal medical history. RESULTS: Thirty-eight percent were overweight (BMI > or = 95th percentile) and 22% at risk for overweight (BMI 85-94th percentile). Thirty-four percent had TSF > or = 90th percentile and 29% had SSSF > or = 90th percentile. Fifty-one percent of males and 70% of females had body fat > 30%. Compared to their nonoverweight counterparts, overweight youth had significantly higher systolic blood pressure (111.4 +/- 1.3 vs. 104.5 +/- 0.9 mm Hg, p < 0.0001). Among children younger than 11 years, overweight was associated with onset of adrenarche (23% vs. 10%, p = 0.01). Participation in one or more sports teams was negatively correlated with overweight) p = 0.04). DISCUSSION: The prevalence of overweight and at risk for overweight in this sample was twice the national average for U.S. children and 1.7 times greater than that of Mexican American children in national surveys. Overweight was associated with advanced pubertal development, high body fat, elevated blood pressure, and decreased sports participation. PMID- 15340114 TI - Definitions of night eating in adolescent girls. AB - OBJECTIVE: To describe the prevalence of night eating in a community cohort of black and white girls, using different definitions of night eating as described in the literature. RESEARCH METHODS AND PROCEDURES: Three-day food diaries collected as part of the National Growth and Health Study were examined to identify episodes of night eating, which was defined in five different ways: eating >25% of daily caloric intake after the last evening meal, eating >25% of daily caloric intake after 7 pm, eating >50% of daily caloric intake after the last evening meal, eating >50% of daily caloric intake after 7 pm, or eating between 11 pm and 4:59 am. RESULTS: Frequency of night eating varied tremendously depending on how the behavior was defined. For the least restrictive definition (>25% of total intake after last meal), 50% to 70% of girls reported one night eating event; for the most restrictive (>50% of total intake after last meal), only 1.5% of 11-year-old girls' diaries and 3.5% of 19-year-old girls' diaries contained a night eating event. The frequency of night eating decreased dramatically (typically by a factor of 10) if the inclusion criteria required multiple night eating events in a given week. DISCUSSION: A standard definition of night eating behavior is needed to advance the field. An agreed-on operationalized definition that includes time of day, amount of calories consumed, and a frequency criterion would enable cross-study comparisons and encourage the examination of developmental and clinical considerations of night eating behavior. PMID- 15340115 TI - Association between low-molecular weight apolipoprotein(a) isoforms and obesity in Italian women. AB - OBJECTIVE: Low-molecular weight (MW) apolipoprotein(a) [apo(a)] isoforms are closely associated with an increased incidence of atherothrombotic disease, prevalence of which is higher in obese individuals, particularly in women. The hypothesis of this study was to assess whether there are differences in the distribution of apo(a) phenotypes between obese patients and healthy controls. RESEARCH METHODS AND PROCEDURES: One hundred three obese Italian women (BMI > or = 30.0 kg/m2) were enrolled in the study, and apo(a) phenotyping was performed in all subjects. The prevalence of low-MW apo(a) isoforms, detected in plasma samples of our obese women, was compared with that found in a control group of 84 normal-weight, never-obese (BMI < 25.0 kg/m(2)), age-matched women. RESULTS: The distribution of apo(a) isoforms in the population of obese women was significantly different from that found in normal-weight female subjects. In particular, the percentage of subjects in the obese group with at least one apo(a) isoform of low MW was significantly higher than that in the control group (51.4% vs. 32.1%, p = 0.0079). DISCUSSION: Our results seem to suggest the possibility that small-sized apo(a) isoforms may be used together with other traditional risk factors to better assess the overall predisposition to atherothrombotic disease in obese women. PMID- 15340116 TI - Identification and characterization of single-nucleotide polymorphisms in MCH-R1 and MCH-R2. AB - OBJECTIVE: To identify and functionally characterize single-nucleotide polymorphisms (SNPs) in melanin-concentrating hormone (MCH)-R1 and -R2. RESEARCH METHODS AND PROCEDURES: The entire coding regions and intron/exon splice junction regions of MCH-R1 and MCH-R2 were sequenced from anonymous white (n=45) and African-American (n=46) individuals. DNA was analyzed, and SNPs were identified using Phred, Phrap, and Consed software. DNA constructs containing MCH-R1 and MCH R2 SNPs were generated and expressed in CHO cells. The effect of the SNPs in MCH R1 and MCH-R2 were assessed in receptor binding assays and functional assays measuring changes in intracellular cAMP and Ca2+ levels. RESULTS: We identified 12 SNPs in the MCH-R1 gene. Two of these SNPs are in coding regions, and one produces an arginine-for-glycine substitution at residue 34 in the MCH-R1 sequence. This SNP is present at a minor allele frequency of 15% in the African American population tested in this study. We identified eight SNPs in the MCH-R2 gene. Four of these SNPs are in coding regions, and two produce amino acid substitutions. Lysine substitutes for arginine at residue 63 of the African American population, and glutamine substitutes for arginine at residue 152 in whites (minor allele frequency of 2% for both SNPs). No changes in receptor binding or functional signaling were observed with the SNP mutations in MCH-R1 or MCH-R2. DISCUSSION: These data indicate that potential therapeutics designed to act at the MCH receptor are unlikely to have altered effects in subpopulations that express variant forms of MCH-R1 or MCH-R2. PMID- 15340117 TI - Dramatic rise in overweight and obesity in adult filipino women and risk of hypertension. AB - OBJECTIVE: To assess trends in BMI of adult Filipino women over a 16-year period of rapid socioeconomic change; to identify factors associated with those trends; and to estimate the risk of hypertension associated with overweight, obesity, and high waist-to-hip ratio (WHR). RESEARCH METHODS AND PROCEDURES: Women from randomly selected urban and rural communities of Metro Cebu, Philippines were recruited during a 1983 to 1984 index pregnancy, then followed prospectively for 16 years. Overweight and obesity were defined using BMI cut-off points of 25 and 30, respectively. The analysis sample included women 15 to 45 years of age when measured 4 months postpartum. Weight change in subsequent intervals from 1985 to 1999 was modeled using linear regression. The relationship of BMI and WHR to risk of hypertension in the last survey was modeled using logistic regression. RESULTS: The prevalence of overweight and obesity combined increased nearly 6 fold from approximately 6% in 1983 to 1984 to 35% in 1998 to 1999. Weight gain was positively associated with urban residence, improved socioeconomic status, fewer pregnancies and months of lactation, and more away-from-home work hours. Risk of hypertension was independently elevated by high WHR and overweight/obesity. DISCUSSION: The dramatic trend of increasing overweight and obesity in this sample of women represents a serious health concern, especially in light of the strong association of excess weight, particularly in the truncal region, to risk of hypertension. PMID- 15340118 TI - Valuing the benefits of weight loss programs: an application of the discrete choice experiment. AB - OBJECTIVE: Obesity is a leading health threat. Determination of optimal therapies for long-term weight loss remains a challenge. Evidence suggests that successful weight loss depends on the compliance of weight loss program participants with their weight loss efforts. Despite this, little is known regarding the attributes influencing such compliance. The purpose of this study was to assess, using a discrete choice experiment (DCE), the relative importance of weight loss program attributes to its participants and to express these preferences in terms of their willingness to pay for them. RESEARCH METHODS: A DCE survey explored the following weight loss program attributes in a sample of 165 overweight adults enrolled in community weight loss programs: cost, travel time required to attend, extent of physician involvement (e.g., none, monthly, every 2 weeks), components (e.g., diet, exercise, behavior change) emphasized, and focus (e.g., group, individual). The rate at which participants were willing to trade among attributes and the willingness to pay for different configurations of combined attributes were estimated using regression modeling. RESULTS: All attributes investigated appeared to be statistically significant. The most important unit change was "program components emphasized" (e.g., moving from diet only to diet and exercise). DISCUSSION: The majority of participants were willing to pay for weight loss programs that reflected their preferences. The DCE tool was useful in quantifying and understanding individual preferences in obesity management and provided information that could help to maximize the efficiency of existing weight loss programs or the design of new programs. PMID- 15340119 TI - Both subcutaneous and visceral adipose tissue correlate highly with insulin resistance in african americans. AB - OBJECTIVE: The contribution of visceral adipose tissue (VAT) to insulin resistance is well-established; however, the role of subcutaneous abdominal adipose tissue (SAT) in insulin resistance remains controversial. Sex may determine which of these two components of abdominal obesity is more strongly related to insulin resistance and its consequences. The aim of this study was to determine whether both VAT and SAT contribute to insulin resistance in African Americans and to examine the effects of sex on this relationship. RESEARCH METHODS AND PROCEDURES: This was a cross-sectional study of 78 nondiabetic African-American volunteers (44 men, 35 women; age 33.8 +/- 7.3 years; BMI 30.9 +/- 7.4 kg/m2). VAT and SAT volumes were measured using serial computerized tomography slices from the dome of the diaphragm to the iliac crest. The insulin sensitivity index (SI) was determined from the minimal model using data obtained from the frequently sampled intravenous glucose tolerance test. RESULTS: In men, both VAT and SAT were negatively correlated with SI (r for both correlations = 0.57; p < 0.01). In women, the correlation coefficient between VAT and SI was 0.50 (p < 0.01) and between SAT and SI was -0.67 (p < 0.01). In women, the correlation coefficient for SI with SAT was significantly greater than the correlation coefficient with VAT (p = 0.02). DISCUSSION: Both SAT and VAT are strongly correlated with insulin resistance in African Americans. For African American women, SAT may have a greater effect than VAT on insulin resistance. PMID- 15340120 TI - Transcriptional profiling of in vitro smooth muscle cell differentiation identifies specific patterns of gene and pathway activation. AB - Mesodermal and epidermal precursor cells undergo phenotypic changes during differentiation to the smooth muscle cell (SMC) lineage that are relevant to pathophysiological processes in the adult. Molecular mechanisms that underlie lineage determination and terminal differentiation of this cell type have received much attention, but the genetic program that regulates these processes has not been fully defined. Study of SMC differentiation has been facilitated by development of the P19-derived A404 embryonal cell line, which differentiates toward this lineage in the presence of retinoic acid and allows selection for cells adopting a SMC fate through a differentiation-specific drug marker. We sought to define global alterations in gene expression by studying A404 cells during SMC differentiation with oligonucleotide microarray transcriptional profiling. Using an in situ 60-mer array platform with more than 20,000 mouse genes derived from the National Institute on Aging clone set, we identified 2,739 genes that were significantly upregulated after differentiation was completed (false-detection ratio <1). These genes encode numerous markers known to characterize differentiated SMC, as well as many unknown factors. We further characterized the sequential patterns of gene expression during the differentiation time course, particularly for known transcription factor families, providing new insights into the regulation of the differentiation process. Changes in genes associated with specific biological ontology-based pathways were evaluated, and temporal trends were identified for functional pathways. In addition to confirming the utility of the A404 model, our data provide a large-scale perspective of gene regulation during SMC differentiation. PMID- 15340121 TI - The gene encoding mouse Muc19: cDNA, genomic organization and relationship to Smgc. AB - We previously demonstrated expression of full-length transcripts for sublingual mucin apoprotein, Muc19, of approximately 24 kb (Fallon MA, Latchney LR, Hand AR, Johar A, Denny PA, Georgel PT, Denny PC, and Culp DJ. Physiol Genomics 14: 95 106, 2003). We now describe the complete sequence and genomic organization of the apomucin encoded by 43 exons. Southern analyses indicate a central exon of approximately 18 kb containing 36 tandem repeats, each encoding 163 residues rich in serine and threonine. Full-length transcripts are an estimated 22,795 bp in length that span 106 kb of genomic DNA. The transcriptional start site is 24 bp downstream of a TATA box and 42 bp upstream of the conceptual translational start codon. The putative apoprotein has an estimated mass of 693.4 kDa and contains 7,524 amino acids (80% serine, threonine, glycine, alanine, and proline). We present a model for rat Muc19 transcripts and compare the conceptually translated Muc19 proteins for mouse, rat, pig, and the 3' end of human Muc19. Conserved among these apoproteins are a signal peptide, a large tandem repeat region, von Willebrand factor type C and D domains, a trypsin inhibitor-like Cys-rich domain, and a COOH-terminal cystine knot-like domain. Southern blot analyses indicate transcripts for Muc19 and Smgc (submandibular gland protein C) are splice variants of a larger gene, Muc19/Smgc. Comparative Northern analyses between the major salivary glands demonstrate highly selective Muc19 expression in neonatal and adult sublingual glands, whereas Smgc is expressed in neonatal submandibular and sublingual glands. Regulation of Muc19/Smgc gene expression is discussed with respect to alternative splicing and mucous cell cytodifferentiation. PMID- 15340122 TI - Subcutaneous furosemide. PMID- 15340123 TI - Geographic region influences pharmacy's dispensing of blood glucose test strips. PMID- 15340124 TI - Recombinant factor VIIa for refractory bleeding following orthotopic heart transplantation. AB - OBJECTIVE: To report a case of successful use of recombinant factor VIIa (rFVIIa) for the treatment of refractory bleeding in a patient undergoing orthotopic heart transplantation. CASE SUMMARY: A 57-year-old white male with idiopathic cardiomyopathy was taken to the operating room for explantation of his left ventricular assist device and orthotopic heart transplantation. He experienced excessive generalized oozing that required transfusions of multiple units of blood products and significant amounts of Cellsaver (washed red blood cells via autotransfusion) without achieving adequate hemostasis. After ruling out any obvious surgical sources of bleeding and attempting to correct all coagulation deficiencies, the clinicians administered rFVIIa 90 microg/kg. The oozing rapidly declined to a negligible level, chest tubes and sternal wires were placed, and the chest was closed. The patient was on minimal inotropic support and was transferred to the intensive care unit in stable condition. DISCUSSION: Cardiac surgery is often associated with significant disruption of the coagulation system, particularly in high-risk patients, such as those undergoing removal of a ventricular assist device and subsequent orthotopic heart transplantation. This can lead to life-threatening bleeding that can require multiple hemostatic agents and significant transfusions to restore hemostasis. Recently, rFVIIa has been utilized as an alternative to massive transfusion for treatment of refractory bleeding in several patient populations, including some cardiac surgery patients. CONCLUSIONS: rFVIIa appears to be a viable option as rescue therapy for treatment of refractory bleeding following orthotopic heart transplantation. Despite the anecdotal success of rFVIIa in this setting, further clinical research is needed. PMID- 15340125 TI - Antidepressants, suicide, and the FDA: a loose association. PMID- 15340126 TI - Antidepressants: an avoidable and solvable controversy. PMID- 15340127 TI - Lumiracoxib does not affect methotrexate pharmacokinetics in rheumatoid arthritis patients. AB - BACKGROUND: Methotrexate and nonsteroidal antiinflammatory drugs are frequently coadministered in the treatment of rheumatoid arthritis (RA). OBJECTIVE: To evaluate the effect of lumiracoxib, a novel cyclooxygenase-2 selective inhibitor, on methotrexate pharmacokinetics and short-term safety in patients with RA. METHODS: This multicenter, randomized, double-blind, placebo-controlled crossover study enrolled 18 patients (mean age 49.1 y) with stable RA. Patients were randomized to receive methotrexate 7.5-15 mg orally once weekly plus either lumiracoxib 400 mg/day or placebo for 7 days. Patients then received the other treatment combination for an additional 7 days. Serial blood and urine were collected for 24 hours after the methotrexate dose on day 1 (methotrexate alone) and days 8 and 15 (combination treatment). RESULTS: Plasma methotrexate pharmacokinetics (AUC(0-t), maximum concentration [C(max)], time to C(max)) and methotrexate protein binding were similar for methotrexate alone (108.0 ng.h/mL, 26.7 ng/mL, 1.5 h, and 57.1%, respectively), methotrexate/lumiracoxib (110.2 ng.h/mL, 27.5 ng/mL, 1.0 h, and 53.7%, respectively), and methotrexate/placebo (101.8 ng.h/mL, 22.6 ng/mL, 1.0 h, and 57.0%, respectively). Similarly, no clinically significant difference was found in the urinary excretion of methotrexate. Mean exposure to the 7-OH metabolite was lower when methotrexate was given with lumiracoxib compared with placebo, shown by a reduction in AUC and C(max), although similar amounts of the metabolite were recovered in urine following both lumiracoxib and placebo. Coadministration of methotrexate and lumiracoxib was well tolerated. CONCLUSIONS: Lumiracoxib had no significant effect on the pharmacokinetics, protein binding, or urinary excretion of coadministered methotrexate in patients with RA. PMID- 15340128 TI - The changing face of antibiotic prescribing: the mutant selection window. AB - OBJECTIVE: To describe the mutant selection window, discuss supporting evidence and limitations, and suggest potential applications for clinical practice. DATA SOURCES: A MEDLINE search (1990-December 2003) of the English-language literature was conducted using the key words antibiotic, antimicrobial, resistance, mutant, selection window, prevention, MPC, and MSW in various combinations. Original investigations and reviews evaluating the mutant selection window, including abstracts and proceedings, were considered for inclusion. Published articles were also cross-referenced, and experts were contacted to locate additional pertinent data. STUDY SELECTION AND DATA EXTRACTION: All data sources identified were evaluated and all information deemed relevant was included. DATA SYNTHESIS: Until recently, physicians have had few ways to preserve antimicrobials from resistance other than by prescribing the agents less often. The mutant selection window hypothesis may modify this paradigm by shifting the focus to dosing strategies that reduce the growth of resistant mutants. Conventional dosing strategies have been formulated on the likelihood of curing an individual patient. Unfortunately, doses that cure patients appear to enrich resistant subpopulations of bacteria, thus promoting resistance. Antimicrobial-pathogen combinations can be identified that minimize mutant selection and cure patients while possibly restricting the progression of resistance. CONCLUSIONS: The mutant selection window hypothesis provides a framework for considering the contribution of dosing to resistance, and it offers ideas for restricting the enrichment of resistant mutants and antimicrobial resistance. PMID- 15340129 TI - Urinary tract infections during pregnancy. AB - OBJECTIVE: To provide a comprehensive review of urinary tract infections (UTIs) during pregnancy. All aspects of UTIs, including epidemiology, pathogenesis, resistance, clinical features, diagnosis, treatment, and prevention, were reviewed. DATA SOURCES: MEDLINE (1966-August 2003) and Cochrane Library searches were performed using the key search terms urinary tract infection, pyelonephritis, cystitis, asymptomatic bacteriuria, and resistance. STUDY SELECTION AND DATA EXTRACTION: All article abstracts were evaluated for relevance. Only articles pertaining to pregnancy were included. The majority of published literature were review articles; the number of original clinical studies was limited. DATA SYNTHESIS: UTIs are the most common bacterial infections during pregnancy. They are characterized by the presence of significant bacteria anywhere along the urinary tract. Pyelonephritis is the most common severe bacterial infection that can lead to perinatal and maternal complications including premature delivery, infants with low birth weight, fetal mortality, preeclampsia, pregnancy-induced hypertension, anemia, thrombocytopenia, and transient renal insufficiency. Enterobacteriaceae account for 90% of UTIs. The common antibiotics used are nitrofurantoin, cefazolin, cephalexin, ceftriaxone, and gentamicin. CONCLUSIONS: Therapeutic management of UTIs in pregnancy requires proper diagnostic workup and thorough understanding of antimicrobial agents to optimize maternal outcome, ensure safety to the fetus, and prevent complications that lead to significant morbidity and mortality in both the fetus and the mother. PMID- 15340130 TI - Acute onset of pancreatitis with concomitant use of tenofovir and didanosine. AB - OBJECTIVE: To report a case of pancreatitis associated with the combined use of didanosine and tenofovir. CASE SUMMARY: A 51-year-old white man with HIV was initiated on antiretroviral therapy with didanosine 250 mg/day, tenofovir 300 mg/day, lamivudine 300 mg/day, stavudine 60 mg/day, and efavirenz 600 mg/day. Didanosine was prescribed at a reduced dosage due to the known interaction with tenofovir. Despite this dosage adjustment, the patient developed acute pancreatitis 10 weeks after antiretrovirals were initiated. Pancreatitis resolved spontaneously after antiretroviral discontinuation. DISCUSSION: Our report of didanosine-induced pancreatitis secondary to concurrent use with tenofovir is the third reported case that utilized a reduced didanosine dosage. Five previous pancreatitis reports have been described using full-strength didanosine with tenofovir. The exact mechanism of action for this interaction is unknown. Utilizing the Naranjo probability scale to assess causality, a possible adverse drug reaction was determined. CONCLUSIONS: Tenofovir and didanosine may be used cautiously in antiretroviral combination therapy. Reduced didanosine dosage (250 mg) should be used to reduce serum didanosine concentrations and subsequent toxicities. Practitioners should be aware that a significant drug interaction with resulting pancreatitis may occur even when a reduced dosage is prescribed. PMID- 15340131 TI - Losartan for treatment of psychogenic polydipsia. PMID- 15340132 TI - Impact of fluconazole administration on outcomes in critically ill patients. AB - BACKGROUND: Serious infections caused by Candida spp. are an increasingly important cause of morbidity and mortality in critically ill patients. It is unclear which patients will benefit from therapy and at what point to institute treatment. OBJECTIVE: To evaluate the impact of administration of fluconazole therapy in critically ill trauma patients on mortality, length of hospital stay, incidence of deep-seated fungal infection, and positive fungal cultures from any site. METHODS: We conducted a retrospective, matched case-control study of 116 critically ill surgical trauma patients who did or did not receive fluconazole. Patients were followed until hospital discharge or death. A consecutive sample of 58 patients who received fluconazole was selected. A parallel group of patients was evaluated, from which 58 were matched with fluconazole-treated patients based on age (+/- 5 y), gender, and APACHE II score (+/- 3). RESULTS: The groups of patients were well matched, with the exception of central venous catheter placement and broad-spectrum antibiotic use. We found no difference between groups in hospital mortality (21% vs 26%; p = 0.661) or incidence of deep-seated fungal infection (0% vs 2%; p = NS). However, patients receiving fluconazole had a significantly longer stay in both the intensive care unit (ICU) (18 +/- 13 vs 7 +/- 11 days; p < 0.001) and hospital (25 +/- 15 vs 9 +/- 11 days; p < 0.001). Fluconazole patients were significantly more likely to have Candida cultured from sites associated with colonization (43% vs 2%; p < 0.001), possibly explaining why they received fluconazole. CONCLUSIONS: We were unable to detect a benefit from use of fluconazole in our surgical trauma patient population. Isolation of Candida from the mouth or throat alone, in the absence of correlating clinical signs of infection, should not lead to initiation of fluconazole therapy. Fluconazole use should be reserved for carefully selected patients in the trauma ICU setting. PMID- 15340133 TI - Micafungin. AB - OBJECTIVE: To review the pharmacology, mycology, chemistry, in vitro susceptibility, pharmacokinetics, clinical efficacy, safety, tolerability, dosage, and administration of micafungin, an echinocandin antifungal agent. DATA SOURCES: A MEDLINE search, restricted to English language, was conducted from 1978 to November 2003. Supplementary sources included program abstracts from the Interscience Conference on Antimicrobial Agents and Chemotherapy and the Infectious Diseases Society of America from 1996 to 2003 and information available through the manufacturer's Web site. STUDY SELECTION AND DATA EXTRACTION: In vitro and preclinical studies, as well as Phase II and III clinical trials, were evaluated to summarize the clinical efficacy and safety of micafungin. All published and unpublished trials and abstracts citing micafungin were selected. DATA SYNTHESIS: Micafungin has shown in vitro activity against many yeasts and a variety of molds. Micafungin can be administered only parenterally. Efficacy has been illustrated in open noncomparative studies of esophageal candidiasis in HIV-infected patients and in comparative trials as antifungal prophylaxis in patients undergoing hematopoietic stem-cell transplantation. Adverse events appear mild and limited; the most commonly reported adverse events include hyperbilirubinemia, nausea, and diarrhea. CONCLUSIONS: Micafungin has activity against Aspergillus spp. and a variety of Candida spp., including azole-resistant strains. Micafungin demonstrates efficacy in the treatment of esophageal candidiasis in HIV-infected patients and appears superior to fluconazole as antifungal prophylaxis in patients undergoing hematopoietic stem-cell transplantation. Based on case reports and in vitro efficacy, micafungin may prove to be a clinically useful agent in the treatment of other fungal diseases; however, these indications await the results of clinical trials. PMID- 15340135 TI - Comment: therapy switching in patients receiving long-acting opioids. PMID- 15340137 TI - Topologically driven swelling of a polymer loop. AB - Numerical studies of the average size of trivially knotted polymer loops with no excluded volume were undertaken. Topology was identified by Alexander and Vassiliev degree 2 invariants. Probability of a trivial knot, average gyration radius, and probability density distributions as functions of gyration radius were generated for loops of up to N = 3,000 segments. Gyration radii of trivially knotted loops were found to follow a power law similar to that of self-avoiding walks consistent with earlier theoretical predictions. PMID- 15340136 TI - Isolation of Mycobacterium tuberculosis mutants defective in the arrest of phagosome maturation. AB - Mycobacterium tuberculosis resides within the phagocytes of its host. It ensures its continued survival through arresting the normal maturation of its phagosome, which is retained within the early endosomal system of the macrophage. Although individual bacterial components have been shown to modulate phagosome biogenesis, the mechanism(s) active in live, intact bacteria remain elusive. We have developed a genetic screen that facilitates the isolation of mutants defective in arresting the maturation of their phagosomes. Macrophages were incubated with iron-dextran that was chased into lysosomes. The cells were subsequently infected with M. tuberculosis from a library of transposon-mutagenized bacteria. After four rounds of enrichment, the majority of mutants isolated were unable to prevent acidification of their phagosomes and were attenuated for intracellular survival. The genes affected range in function from those with no known homologues to putative transporters and lipid synthesis enzymes. Further characterization of these bacteria is needed. In addition to clarifying the processes active in modulation of phagosome biogenesis by M. tuberculosis, this screen may be applicable to other pathogens that restrict the maturation of their phagosome. PMID- 15340138 TI - Analysis of Rhizobium etli and of its symbiosis with wild Phaseolus vulgaris supports coevolution in centers of host diversification. AB - Common beans (Phaseolus vulgaris) comprise three major geographic genetic pools, one in Mexico, Central America, and Colombia, another in the southern Andes, and a third in Ecuador and northern Peru. Species Rhizobium etli is the predominant rhizobia found symbiotically associated with beans in the Americas. We have found polymorphism in the common nodulation gene nodC among R. etli strains from a wide range of geographical origins, which disclosed three nodC types. The different nodC alleles in American strains show varying predominance in their regional distributions in correlation with the centers of bean genetic diversification (BD centers). By cross-inoculating wild common beans from the three BD centers with soils from Mexico, Ecuador, Bolivia, and Northwestern Argentina, the R. etli populations from nodules originated from Mexican soil again showed allele predominance that was opposite to those originated from Bolivian and Argentinean soil, whereas populations from Ecuadorian soil were intermediate. These results also indicated that the preferential nodulation of beans by geographically related R. etli lineages was independent of the nodulating environment. Coinoculation of wild common beans from each of the three BD centers with an equicellular mixture of R. etli strains representative of the Mesoamerican and southern Andean lineages revealed a host-dependent distinct competitiveness: beans from the Mesoamerican genetic pool were almost exclusively nodulated by strains from their host region, whereas nodules of beans from the southern Andes were largely occupied by the geographically cognate R. etli lineages. These results suggest coevolution in the centers of host genetic diversification. PMID- 15340139 TI - Ribosomal protein S1 binds mRNA and tmRNA similarly but plays distinct roles in translation of these molecules. AB - Ribosomes stalled during protein synthesis can be rescued by tmRNA, which acts first as a tRNA and then as an mRNA to direct addition of a C-terminal degradation tag to the nascent polypeptide. Ribosomal protein S1 binds tmRNA, but its functional role in tmRNA-mediated tagging is uncertain. To probe interactions between S1 and tmRNA, truncated variants missing one or more of the six contiguous S1 domains were studied. The third S1 domain (R1) plays a critical role in binding tmRNA and mRNA but requires additional N- or C-terminal S1 domains. The binding of S1 and its fragments to tmRNA and mRNA is positively cooperative, and the essential role of the R1 domain may be to mediate protein protein interactions. Overproduction of N-terminal fragments of S1 in Escherichia coli displaces endogenous S1 from ribosomes, inhibits general protein synthesis, and slows growth but causes little if any disruption of tmRNA-mediated tagging. Moreover, tagging of proteins translated from model mRNAs with either no or an increased requirement for S1 is indistinguishable. These results raise the possibility that S1 plays little or no role in tmRNA-mediated tagging. PMID- 15340140 TI - Sperm-specific protein kinase A catalytic subunit Calpha2 orchestrates cAMP signaling for male fertility. AB - An unusual cAMP signaling system mediates many of the events that prepare spermatozoa to meet the egg. Its components include the atypical, bicarbonate stimulated, sperm adenylyl cyclase and a cAMP-dependent protein kinase (PKA) with the unique catalytic subunit termed Calpha(2) or C(s). We generated mice that lack Calpha(2) to determine its importance in the events downstream of cAMP production. Male Calpha(2) null mice produce normal numbers of sperm that swim spontaneously in vitro. Thus, Calpha(2) has no required role in formation of a functional flagellum or the initiation of motility. In contrast, we find that Calpha(2) is required for bicarbonate to speed the flagellar beat and facilitate Ca(2+) entry channels. In addition, Calpha(2) is needed for the protein tyrosine phosphorylation that occurs late in the sequence of sperm maturation and for a negative feedback control of cAMP production, revealed here. Consistent with these specific defects in several important sperm functions, Calpha(2) null males are infertile despite normal mating behavior. These results define several crucial roles of PKA in sperm cell biology, bringing together both known and unique PKA-mediated events that are necessary for male fertility. PMID- 15340141 TI - Biography of Charles R. Gallistel. PMID- 15340142 TI - Biography of Barry S. Coller. PMID- 15340143 TI - Proendocrine genes coordinate the pancreatic islet differentiation program in vitro. AB - In the developing pancreas, the basic helix-loop-helix (bHLH) protein Neurogenin3 (Ngn3) specifies which precursor cells ultimately will become endocrine cells and initiates the islet differentiation program. NeuroD1, a closely related bHLH protein and a downstream target of Ngn3, maintains the differentiation program initiated by Ngn3. We have developed an in vitro model of Ngn3-dependent differentiation by infecting pancreatic duct cell lines with an Ngn3-expressing adenovirus. We found that both Ngn3 and its downstream target NeuroD1 activated the islet differentiation program in these cells by inducing the expression of genes with early roles in the differentiation cascade, as well as genes characteristic of fully differentiated islet cells. Induction of these genes, as exemplified by the insulin1 gene, involved alteration of the local chromatin structure. Interestingly, the subsets of genes activated by Ngn3 and NeuroD1 were not completely overlapping, indicating that these two bHLH proteins serve specific functions in the development of the endocrine pancreas. In addition, microarray gene expression analysis identified a previously uncharacterized group of Ngn3-induced genes with potentially important roles in islet development and function. These studies demonstrate how Ngn3 initiates islet differentiation and provide us with a model for testing methods for producing islet cells for people with diabetes. PMID- 15340144 TI - Homeostatic regulation of dopaminergic neurons without dopamine. PMID- 15340145 TI - Cycling antibiotics may not be good for your health. PMID- 15340146 TI - Exploring the O-GlcNAc proteome: direct identification of O-GlcNAc-modified proteins from the brain. AB - The covalent modification of intracellular proteins by O-linked beta-N acetylglucosamine (O-GlcNAc) is emerging as a crucial regulatory posttranslational modification akin to phosphorylation. Numerous studies point to the significance of O-GlcNAc in cellular processes such as nutrient sensing, protein degradation, and gene expression. Despite its importance, the breadth and functional roles of O-GlcNAc are only beginning to be elucidated. Advances in our understanding will require the development of new strategies for the detection and study of O-GlcNAc-modified proteins in vivo. Herein we report the direct, high-throughput analysis of O-GlcNAc-glycosylated proteins from the mammalian brain. The proteins were identified by using a chemoenzymatic approach that exploits an engineered galactosyltransferase enzyme to selectively label O-GlcNAc proteins with a ketone-biotin tag. The tag permits enrichment of low-abundance O GlcNAc species from complex mixtures and localization of the modification to short amino acid sequences. Using this approach, we discovered 25 O-GlcNAc glycosylated proteins from the brain, including regulatory proteins associated with gene expression, neuronal signaling, and synaptic plasticity. The functional diversity represented by this set of proteins suggests an expanded role for O GlcNAc in regulating neuronal function. Moreover, the chemoenzymatic strategy described here should prove valuable for identifying O-GlcNAc-modified proteins in various tissues and facilitate studies of the physiological significance of O GlcNAc across the proteome. PMID- 15340147 TI - O2 activation by binuclear Cu sites: noncoupled versus exchange coupled reaction mechanisms. AB - Binuclear Cu proteins play vital roles in O(2) binding and activation in biology and can be classified into coupled and noncoupled binuclear sites based on the magnetic interaction between the two Cu centers. Coupled binuclear Cu proteins include hemocyanin, tyrosinase, and catechol oxidase. These proteins have two Cu centers strongly magnetically coupled through direct bridging ligands that provide a mechanism for the 2-electron reduction of O(2) to a mu-eta(2):eta(2) side-on peroxide bridged Cu(II)(2)(O(2)(2-)) species. This side-on bridged peroxo Cu(II)(2) species is activated for electrophilic attack on the phenolic ring of substrates. Noncoupled binuclear Cu proteins include peptidylglycine alpha hydroxylating monooxygenase and dopamine beta-monooxygenase. These proteins have binuclear Cu active sites that are distant, that exhibit no exchange interaction, and that activate O(2) at a single Cu center to generate a reactive Cu(II)/O(2) species for H-atom abstraction from the C-H bond of substrates. O(2) intermediates in the coupled binuclear Cu enzymes can be trapped and studied spectroscopically. Possible intermediates in noncoupled binuclear Cu proteins can be defined through correlation to mononuclear Cu(II)/O(2) model complexes. The different intermediates in these two classes of binuclear Cu proteins exhibit different reactivities that correlate with their different electronic structures and exchange coupling interactions between the binuclear Cu centers. These studies provide insight into the role of exchange coupling between the Cu centers in their reaction mechanisms. PMID- 15340148 TI - Unique role of CD4+CD62L+ regulatory T cells in the control of autoimmune diabetes in T cell receptor transgenic mice. AB - Converging experimental evidence indicates that CD4(+) regulatory T cells control progression of autoimmune insulitis in nonobese diabetic (NOD) mice. Here, we studied the nature of these regulatory T cells and their mode of action in diabetes-prone NOD Rag(-/-) or severe combined immunodeficient (SCID) mice harboring a transgenic T cell receptor derived from the diabetogenic T cell clone BDC2.5. We first show that diabetes onset is prevented in such mice by infusion of polyclonal CD4(+) T cells expressing L-selectin (CD62L) but not prevented or only marginally prevented by CD4(+)CD25(+) T cells. Similarly, we found with a cotransfer model that CD4(+)CD62L(+) T cells but not CD4(+)CD25(+) T cells inhibited diabetes transfer into NOD SCID recipients by transgenic NOD BDC2.5 SCID cells. Unexpectedly, cotransfer of transgenic NOD BDC2.5 SCID cells and spleen cells from WT diabetic NOD mice did not induce diabetes, whereas each individual population did so. Data are presented arguing for the role of CD4(+)CD62L(+) T cells present within the polyclonal diabetogenic population in mediating this apparently paradoxical effect. Collectively, these data confirm the central role of CD4(+)CD62L(+) regulatory T cells in controlling disease onset in a well defined transgenic model of autoimmune diabetes and suggest the intervention of homeostatic mechanisms as part of their mode of action. PMID- 15340149 TI - RELMbeta/FIZZ2 is a goblet cell-specific immune-effector molecule in the gastrointestinal tract. AB - Gastrointestinal (GI) nematode infections are an important public health and economic concern. Experimental studies have shown that resistance to infection requires CD4(+) T helper type 2 (Th2) cytokine responses characterized by the production of IL-4 and IL-13. However, despite >30 years of research, it is unclear how the immune system mediates the expulsion of worms from the GI tract. Here, we demonstrate that a recently described intestinal goblet cell-specific protein, RELMbeta/FIZZ2, is induced after exposure to three phylogenetically distinct GI nematode pathogens. Maximal expression of RELMbeta was coincident with the production of Th2 cytokines and host protective immunity, whereas production of the Th1 cytokine, IFN-gamma, inhibited RELMbeta expression and led to chronic infection. Furthermore, whereas induction of RELMbeta was equivalent in nematode-infected wild-type and IL-4-deficient mice, IL-4 receptor-deficient mice showed minimal RELMbeta induction and developed persistent infections, demonstrating a direct role for IL-13 in optimal expression of RELMbeta. Finally, we show that RELMbeta binds to components of the nematode chemosensory apparatus and inhibits chemotaxic function of a parasitic nematode in vitro. Together, these results suggest that intestinal goblet cell-derived RELMbeta may be a novel Th2 cytokine-induced immune-effector molecule in resistance to GI nematode infection. PMID- 15340150 TI - Type II platelet-activating factor-acetylhydrolase is essential for epithelial morphogenesis in Caenorhabditis elegans. AB - Type II platelet-activating factor-acetylhydrolase [PAF-AH (II)] is an N myristoylated enzyme that contains a lipase/esterase catalytic motif and selectively hydrolyzes the sn-2 acetyl ester of PAF and other short-chain acyl groups attached to phosphoglycerides. However, the physiological role of this enzyme remains to be elucidated. PAF-AH (II) is conserved in a variety of species ranging from a simple multicellular organism, Caenorhabditis elegans, to mammals. C. elegans possesses two homologous PAF-AH (II) genes, named paf-1 and paf-2. In this study, we generated these two loss-of-function mutants to elucidate the in vivo PAF-AH (II) function. Surprisingly, mutants of paf-2, a major isoform of C. elegans PAF-AH (II)s, exhibits gross defects in epithelial sheet formation, resulting in unsuccessful subsequent morphogenesis with complete penetrance. Moreover, paf-2 RNA interference worms show a variable abnormal morphology, including ectopic protrusions and a lumpy shape at the late embryonic and early larval stages due to epithelial organization defects. Consistent with these phenotypes, PAF-AH (II) is predominantly expressed in epithelial cells of C. elegans. This study demonstrates that PAF-AH (II) is essential for epithelial morphogenesis. PMID- 15340151 TI - Real-time imaging of single nerve cell apoptosis in retinal neurodegeneration. AB - Apoptotic nerve cell death is implicated in the pathogenesis of several devastating neurodegenerative conditions, including glaucoma and Alzheimer's and Parkinson's diseases. We have devised a noninvasive real-time imaging technique using confocal laser-scanning ophthalmoscopy to visualize single nerve cell apoptosis in vivo, which allows longitudinal study of disease processes that has not previously been possible. Our method utilizes the unique optical properties of the eye, which allow direct microscopic observation of nerve cells in the retina. We have been able to image changes occurring in nerve cell apoptosis over hours, days, and months and show that effects depend on the magnitude of the initial apoptotic inducer in several models of neurodegenerative disease in rat and primate. This technology enables the direct observation of single nerve cell apoptosis in experimental neurodegeneration, providing the opportunity for detailed investigation of fundamental disease mechanisms and the evaluation of interventions with potential clinical applications, together with the possibility of taking this method through to patients. PMID- 15340152 TI - Measurement of anisotropic energy transport in flowing polymers by using a holographic technique. AB - Almost no experimental data exist to test theories for the nonisothermal flow of complex fluids. To provide quantitative tests for newly proposed theories, we have developed a holographic grating technique to study energy transport in an amorphous polymer melt subject to flow. Polyisobutylene with weight-averaged molecular mass of 85 kDa is sheared at a rate of 10 s(-1), and all nonzero components of the thermal conductivity tensor are measured as a function of time, after cessation. Our results are consistent with proposed generalizations to the energy balance for microstructural fluids, including a generalized Fourier's law for anisotropic media. The data are also consistent with a proposed stress thermal rule for amorphous polymer melts. Confirmation of the universality of these results would allow numerical modelers to make quantitative predictions for the nonisothermal flow of polymer melts. PMID- 15340153 TI - Universality in intermediary metabolism. AB - We analyze the stoichiometry, energetics, and reaction concentration dependence of the reductive tricarboxylic acid (rTCA) cycle as a universal and possibly primordial metabolic core. The rTCA reaction sequence is a network-autocatalytic cycle along the relaxation pathway for redox couples in nonequilibrium reducing environments, which provides starting organic compounds for the synthesis of all major classes of biomolecules. The concentration dependence of its reactions suggests it as a precellular bulk process. We propose that rTCA is statistically favored among competing redox relaxation pathways under early-earth conditions and that this feature drove its emergence and also accounts for its evolutionary robustness and universality. The ability to enhance the rate of core reactions creates an energetic basis for selection of subsequent layers of biological complexity. PMID- 15340154 TI - Membrane lipids of symbiotic algae are diagnostic of sensitivity to thermal bleaching in corals. AB - Over the past three decades, massive bleaching events of zooxanthellate corals have been documented across the range of global distribution. Although the phenomenon is correlated with relatively small increases in sea-surface temperature and enhanced light intensity, the underlying physiological mechanism remains unknown. In this article we demonstrate that thylakoid membrane lipid composition is a key determinate of thermal-stress sensitivity in symbiotic algae of cnidarians. Analyses of thylakoid membranes reveal that the critical threshold temperature separating thermally tolerant from sensitive species of zooxanthellae is determined by the saturation of the lipids. The lipid composition is potentially diagnostic of the differential nature of thermally induced bleaching found in scleractinian corals. Measurements of variable chlorophyll fluorescence kinetic transients indicate that thermally damaged membranes are energetically uncoupled but remain capable of splitting water. Consequently, a fraction of the photosynthetically produced oxygen is reduced by photosystem I through the Mehler reaction to form reactive oxygen species, which rapidly accumulate at high irradiance levels and trigger death and expulsion of the endosymbiotic algae. Differential sensitivity to thermal stress among the various species of Symbiodinium seems to be distributed across all clades. A clocked molecular phylogenetic analysis suggests that the evolutionary history of symbiotic algae in cnidarians selected for a reduced tolerance to elevated temperatures in the latter portion of the Cenozoic. PMID- 15340155 TI - Robustness properties of circadian clock architectures. AB - Robustness, a relative insensitivity to perturbations, is a key characteristic of living cells. However, the specific structural characteristics that are responsible for robust performance are not clear, even in genetic circuits of moderate complexity. Formal sensitivity analysis allows the investigation of robustness and fragility properties of mathematical models representing regulatory networks, but it yields only local properties with respect to a particular choice of parameter values. Here, we show that by systematically investigating the parameter space, more global properties linked to network structure can be derived. Our analysis focuses on the genetic oscillator responsible for generating circadian rhythms in Drosophila as a prototypic dynamical cellular system. Analysis of two mathematical models of moderate complexity shows that the tradeoff between robustness and fragility is largely determined by the regulatory structure. Rank-ordered sensitivities, for instance, allow the correct identification of protein phosphorylation as an influential process determining the oscillator's period. Furthermore, sensitivity analysis confirms the theoretical insight that hierarchical control might be important for achieving robustness. The complex feedback structures encountered in vivo, however, do not seem to enhance robustness per se but confer robust precision and adjustability of the clock while avoiding catastrophic failure. PMID- 15340156 TI - The immunoglobulin family member dendrite arborization and synapse maturation 1 (Dasm1) controls excitatory synapse maturation. AB - In the developing mammalian brain, a large fraction of excitatory synapses initially contain only N-methyl-D-aspartate receptor and thus are "silent" at the resting membrane potential. As development progresses, synapses acquire alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPA-Rs). Although this maturation of excitatory synapses has been well characterized, the molecular basis for this developmental change is not known. Here, we report that dendrite arborization and synapse maturation 1 (Dasm1), an Ig superfamily member, controls excitatory synapse maturation. Dasm1 is localized at the excitatory synapses. Suppression of Dasm1 expression by using RNA interference or expression of dominant negative deletion mutants of Dasm1 in hippocampal neurons at late developmental stage specifically impairs AMPA-R-mediated, but not N-methyl-D aspartate receptor-mediated, synaptic transmission. The ability of Dasm1 to regulate synaptic AMPA-Rs requires its intracellular C-terminal PDZ domain binding motif, which interacts with two synaptic PDZ domain-containing proteins involved in spine/synapse maturation, Shank and S-SCAM. Moreover, expression of dominant negative deletion mutants of Dasm1 leads to more immature silent synapses. These results suggest that Dasm1, as a transmembrane molecule, likely provides a link to bridge extracellular signals and intracellular signaling complexes in controlling excitatory synapse maturation. PMID- 15340157 TI - Control of dendrite arborization by an Ig family member, dendrite arborization and synapse maturation 1 (Dasm1). AB - Development of both dendrites and axons is important for the formation of neuronal circuits, because dendrites receive information and the axon is responsible for sending signals. In the past decade, extensive studies have revealed many molecules underlying axonal outgrowth and pathfinding. In contrast, much less is known about the molecular mechanisms that control dendrite development. Here we report the identification of an evolutionarily conserved Ig superfamily member, dendrite arborization and synapse maturation 1 (Dasm1), which plays a critical role in dendrite development. Dasm1 contains five Ig domains and two fibronectin III domains in the extracellular N terminus, a single transmembrane domain, and an intracellular C-terminal tail with a type I PDZ domain binding motif at the end. It is highly expressed in the brain and localized at the dendrites. Suppression of Dasm1 expression in hippocampal neurons via RNA interference or expression of Dasm1 without its cytoplasmic tail specifically impairs dendrite, but not axon, outgrowth. Together with its orthologues in other species, Dasm1 defines a family of molecules likely involved specifically in dendrite arborization. PMID- 15340158 TI - Changes in connectivity profiles define functionally distinct regions in human medial frontal cortex. AB - A fundamental issue in neuroscience is the relation between structure and function. However, gross landmarks do not correspond well to microstructural borders and cytoarchitecture cannot be visualized in a living brain used for functional studies. Here, we used diffusion-weighted and functional MRI to test structure-function relations directly. Distinct neocortical regions were defined as volumes having similar connectivity profiles and borders identified where connectivity changed. Without using prior information, we found an abrupt profile change where the border between supplementary motor area (SMA) and pre-SMA is expected. Consistent with this anatomical assignment, putative SMA and pre-SMA connected to motor and prefrontal regions, respectively. Excellent spatial correlations were found between volumes defined by using connectivity alone and volumes activated during tasks designed to involve SMA or pre-SMA selectively. This finding demonstrates a strong relationship between structure and function in medial frontal cortex and offers a strategy for testing such correspondences elsewhere in the brain. PMID- 15340159 TI - Enhanced neurogenesis in Alzheimer's disease transgenic (PDGF-APPSw,Ind) mice. AB - Neurogenesis continues in the adult brain and is increased in certain pathological states. We reported recently that neurogenesis is enhanced in hippocampus of patients with Alzheimer's disease (AD). We now report that the effect of AD on neurogenesis can be reproduced in a transgenic mouse model. PDGF APP(Sw,Ind) mice, which express the Swedish and Indiana amyloid precursor protein mutations, show increased incorporation of BrdUrd and expression of immature neuronal markers in two neuroproliferative regions: the dentate gyrus and subventricular zone. These changes, consisting of approximately 2-fold increases in the number of BrdUrd-labeled cells, were observed at age 3 months, when neuronal loss and amyloid deposition are not detected. Because enhanced neurogenesis occurs in both AD and an animal model of AD, it seems to be caused by the disease itself and not by confounding clinical factors. As neurogenesis is increased in PDGF-APP(Sw,Ind) mice in the absence of neuronal loss, it must be triggered by more subtle disease manifestations, such as impaired neurotransmission. Enhanced neurogenesis in AD and animal models of AD suggests that neurogenesis may be a compensatory response and that measures to enhance neurogenesis further could have therapeutic potential. PMID- 15340160 TI - Mapping activation and repression domains of the vnd/NK-2 homeodomain protein. AB - A transient transfection assay using Drosophila S2 tissue culture cells and WT and mutant Drosophila vnd/NK-2 homeobox cDNAs was used to localize repression and activation domains of vnd/NK-2 homeodomain protein. A repression domain was identified near the N terminus of vnd/NK-2 homeodomain protein (amino acid residues 154-193), which contains many hydrophobic amino acid residues. The major determinants of the repression domain were shown to be amino acid residues F155, W158, I161, L162, L163, and W166. Truncated protein consisting of the N-terminal repression domain and the DNA-binding homeodomain repressed transcription as efficiently as WT vnd/NK-2 protein. An activation domain was identified between the tinman domain and the homeodomain (amino acid residues 277-543), which consists of a glutamine-rich subdomain and two acidic subdomains. No effect was detected of the tinman domain or the NK-2-specific domain on either activation or repression of a beta-galactosidase reporter gene. PMID- 15340161 TI - Signal peptide prediction based on analysis of experimentally verified cleavage sites. AB - A number of computational tools are available for detecting signal peptides, but their abilities to locate the signal peptide cleavage sites vary significantly and are often less than satisfactory. We characterized a set of 270 secreted recombinant human proteins by automated Edman analysis and used the verified cleavage sites to evaluate the success rate of a number of computational prediction programs. An examination of the frequency of amino acid in the N terminal region of the data set showed a preference of proline and glutamine but a bias against tyrosine. The data set was compared to the SWISS-PROT database and revealed a high percentage of discrepancies with cleavage site annotations that were computationally generated. The best program for predicting signal sequences was found to be SignalP 2.0-NN with an accuracy of 78.1% for cleavage site recognition. The new data set can be utilized for refining prediction algorithms, and we have built an improved version of profile hidden Markov model for signal peptides based on the new data. PMID- 15340162 TI - Optimal bundling of transmembrane helices using sparse distance constraints. AB - We present a two-step approach to modeling the transmembrane spanning helical bundles of integral membrane proteins using only sparse distance constraints, such as those derived from chemical cross-linking, dipolar EPR and FRET experiments. In Step 1, using an algorithm, we developed, the conformational space of membrane protein folds matching a set of distance constraints is explored to provide initial structures for local conformational searches. In Step 2, these structures refined against a custom penalty function that incorporates both measures derived from statistical analysis of solved membrane protein structures and distance constraints obtained from experiments. We begin by describing the statistical analysis of the solved membrane protein structures from which the theoretical portion of the penalty function was derived. We then describe the penalty function, and, using a set of six test cases, demonstrate that it is capable of distinguishing helical bundles that are close to the native bundle from those that are far from the native bundle. Finally, using a set of only 27 distance constraints extracted from the literature, we show that our method successfully recovers the structure of dark-adapted rhodopsin to within 3.2 A of the crystal structure. PMID- 15340163 TI - Simulating evolution by gene duplication of protein features that require multiple amino acid residues. AB - Gene duplication is thought to be a major source of evolutionary innovation because it allows one copy of a gene to mutate and explore genetic space while the other copy continues to fulfill the original function. Models of the process often implicitly assume that a single mutation to the duplicated gene can confer a new selectable property. Yet some protein features, such as disulfide bonds or ligand binding sites, require the participation of two or more amino acid residues, which could require several mutations. Here we model the evolution of such protein features by what we consider to be the conceptually simplest route point mutation in duplicated genes. We show that for very large population sizes N, where at steady state in the absence of selection the population would be expected to contain one or more duplicated alleles coding for the feature, the time to fixation in the population hovers near the inverse of the point mutation rate, and varies sluggishly with the lambda(th) root of 1/N, where lambda is the number of nucleotide positions that must be mutated to produce the feature. At smaller population sizes, the time to fixation varies linearly with 1/N and exceeds the inverse of the point mutation rate. We conclude that, in general, to be fixed in 10(8) generations, the production of novel protein features that require the participation of two or more amino acid residues simply by multiple point mutations in duplicated genes would entail population sizes of no less than 10(9). PMID- 15340164 TI - Prediction of pKa and redox properties in the thioredoxin superfamily. AB - Electrostatic interactions play important roles in diverse biological phenomena controlling the function of many proteins. Polar molecules can be studied with the FDPB method solving the Poisson-Boltzmann equation on a finite difference grid. A method for the prediction of pK(a)s and redox potentials in the thioredoxin superfamily is introduced. The results are compared with experimental pK(a) data where available, and predictions are made for members lacking such data. Studying CxxC motif variation in the context of different background structures permits analysis of contributions to cysteine DeltapK(a)s. The motif itself and the overall framework regulate pK(a) variation. The reported method includes generation of multiple side-chain rotamers for the CxxC motif and is an effective predictive tool for functional pK(a) variation across the superfamily. Redox potential follows the trend in cysteine pK(a) variation, but some residual discrepancy indicates that a pH-independent factor plays a role in determining redox potentials for at least some members of the superfamily. A possible molecular basis for this feature is discussed. PMID- 15340165 TI - Crystallographic identification of Ca2+ and Sr2+ coordination sites in synaptotagmin I C2B domain. AB - Synaptotagmin I has two tandem Ca(2+)-binding C(2) domains, which are essential for fast synchronous synaptic transmission in the central nervous system. We have solved four crystal structures of the C(2)B domain, one of them in the cation free form at 1.50 A resolution, two in the Ca(2+)-bound form at 1.04 A (two bound Ca(2+) ions) and 1.65 A (three bound Ca(2+) ions) resolution and one in the Sr(2+)-bound form at 1.18 A (one bound Sr(2+) ion) resolution. The side chains of four highly conserved aspartic acids (D303, D309, D363, and D365) and two main chain oxygens (M302:O and Y364:O), together with water molecules, are in direct contact with two bound Ca(2+) ions (sites 1 and 2). At higher Ca(2+) concentrations, the side chain of N333 rotates and cooperates with D309 to generate a third Ca(2+) coordination site (site 3). Divalent cation binding sites 1 and 2 in the C(2)B domain were previously identified from NMR NOE patterns and titration studies, supplemented by site-directed mutation analysis. One difference between the crystal and NMR studies involves D371, which is not involved in coordination with any of the identified Ca(2+) sites in the crystal structures, while it is coordinated to Ca(2+) in site 2 in the NMR structure. In the presence of Sr(2+), which is also capable of triggering exocytosis, but with lower efficiency, only one cation binding site (site 1) was occupied in the crystallographic structure. PMID- 15340166 TI - Probing the influence on folding behavior of structurally conserved core residues in P. aeruginosa apo-azurin. AB - The effects on folding kinetics and equilibrium stability of core mutations in the apo-mutant C112S of azurin from Pseudomonas aeruginosa were studied. A number of conserved residues within the cupredoxin family were recognized by sequential alignment as constituting a common hydrophobic core: I7, F15, L33, W48, F110, L50, V95, and V31. Of these, I7, V31, L33, and L50 were mutated for the purpose of obtaining information on the transition state and a potential folding nucleus. In addition, residue V5 in the immediate vicinity of the common core, as well as T52, separate from the core, were mutated as controls. All mutants exhibited a nonlinear dependence of activation free energy of folding on denaturant concentration, although the refolding kinetics of the V31A/C112S mutant indicated that the V31A mutation destabilizes the transition state enough to allow folding via a parallel transition state ensemble. Phi-values could be calculated for three of the six mutants, V31A/C112S, L33A/C112S, and L50A/C112S, and the fractional values of 0.63, 0.33, and 0.50 (respectively) obtained at 0.5 M GdmCl suggest that these residues are important for stabilizing the transition state. Furthermore, a linear dependence of ln k(obs)(H2O) on DeltaG(U-N)(H2O) of the core mutations and the putative involvement of ground-state effects suggest the presence of native-like residual interactions in the denatured state that bias this ensemble toward a folding-competent state. PMID- 15340167 TI - Quantification of helix-helix binding affinities in micelles and lipid bilayers. AB - A theoretical approach for estimating association free energies of alpha-helices in nonpolar media has been developed. The parameters of energy functions have been derived from DeltaDeltaG values of mutants in water-soluble proteins and partitioning of organic solutes between water and nonpolar solvents. The proposed approach was verified successfully against three sets of published data: (1) dissociation constants of alpha-helical oligomers formed by 27 hydrophobic peptides; (2) stabilities of 22 bacteriorhodopsin mutants, and (3) protein-ligand binding affinities in aqueous solution. It has been found that coalescence of helices is driven exclusively by van der Waals interactions and H-bonds, whereas the principal destabilizing contributions are represented by side-chain conformational entropy and transfer energy of atoms from a detergent or lipid to the protein interior. Electrostatic interactions of alpha-helices were relatively weak but important for reproducing the experimental data. Immobilization free energy, which originates from restricting rotational and translational rigid-body movements of molecules during their association, was found to be less than 1 kcal/mole. The energetics of amino acid substitutions in bacteriorhodopsin was complicated by specific binding of lipid and water molecules to cavities created in certain mutants. PMID- 15340168 TI - Crystal structure of Ski8p, a WD-repeat protein with dual roles in mRNA metabolism and meiotic recombination. AB - Ski8p is a WD-repeat protein with an essential role for the Ski complex assembly in an exosome-dependent 3'-to-5' mRNA decay. In addition, Ski8p is involved in meiotic recombination by interacting with Spo11p protein. We have determined the crystal structure of Ski8p from Saccharomyces cerevisiae at 2.2 A resolution. The structure reveals that Ski8p folds into a seven-bladed beta propeller. Mapping sequence conservation and hydrophobicities of amino acids on the molecular surface of Ski8p reveals a prominent site on the top surface of the beta propeller, which is most likely involved in mediating interactions of Ski8p with Ski3p and Spo11p. Mutagenesis combined with yeast two-hybrid and GST pull-down assays identified the top surface of the beta propeller as being required for Ski8p binding to Ski3p and Spo11p. The functional implications for Ski8p function in both mRNA decay and meiotic recombination are discussed. PMID- 15340169 TI - Structure and function of the Gondwanian hemoglobin of Pseudaphritis urvillii, a primitive notothenioid fish of temperate latitudes. AB - The suborder Notothenioidei dominates the Antarctic ichthyofauna. The non Antarctic monotypic family Pseudaphritidae is one of the most primitive families. The characterization of the oxygen-transport system of euryhaline Pseudaphritis urvillii is herewith reported. Similar to most Antarctic notothenioids, this temperate species has a single major hemoglobin (Hb 1, over 95% of the total). Hb 1 has strong Bohr and Root effects. It shows two very uncommon features in oxygen binding: At high pH values, the oxygen affinity is exceptionally high compared to other notothenioids, and subunit cooperativity is modulated by pH in an unusual way, namely the curve of the Hill coefficient is bell-shaped, with values approaching 1 at both extremes of pH. Molecular modeling, electronic absorption and resonance Raman spectra have been used to characterize the heme environment of Hb 1 in an attempt to explain these features, particularly in view of some potentially important nonconservative replacements found in the primary structure. Compared to human HbA, no major changes were found in the structure of the proximal cavity of the alpha-chain of Hb 1, although an altered distal histidyl and heme position was identified in the models of the beta-chain, possibly facilitated by a more open heme pocket due to reduced steric constraints on the vinyl substituent groups. This conformation may lead to the hemichrome form identified by spectroscopy in the Met state, which likely fulfils a potentially important physiological role. PMID- 15340170 TI - Crystal structure of ribosomal protein L27 from Thermus thermophilus HB8. AB - Ribosomal protein L27 is located near the peptidyltransferase center at the interface of ribosomal subunits, and is important for ribosomal assembly and function. We report the crystal structure of ribosomal protein L27 from Thermus thermophilus HB8, which was determined by the multiwavelength anomalous dispersion method and refined to an R-factor of 19.7% (R(free) = 23.6%) at 2.8 A resolution. The overall fold is an all beta-sheet hybrid. It consists of two sets of four-stranded beta-sheets formed around a well-defined hydrophobic core, with a highly positive charge on the protein surface. The structure of ribosomal protein L27 from T. thermophilus HB8 in the RNA-free form is investigated, and its functional roles in the ribosomal subunit are discussed. PMID- 15340171 TI - Alanine-scanning mutagenesis of the beta-sheet region of phage T4 lysozyme suggests that tertiary context has a dominant effect on beta-sheet formation. AB - In general, alpha-helical conformations in proteins depend in large part on the amino acid residues within the helix and their proximal interactions. For example, an alanine residue has a high propensity to adopt an alpha-helical conformation, whereas that of a glycine residue is low. The sequence preferences for beta-sheet formation are less obvious. To identify the factors that influence beta-sheet conformation, a series of scanning polyalanine mutations were made within the strands and associated turns of the beta-sheet region in T4 lysozyme. For each construct the stability of the folded protein was reduced substantially, consistent with removal of native packing interactions. However, the crystal structures showed that each of the mutants retained the beta-sheet conformation. These results suggest that the structure of the beta-sheet region of T4 lysozyme is maintained to a substantial extent by tertiary interactions with the surrounding parts of the protein. Such tertiary interactions may be important in determining the structures of beta-sheets in general. PMID- 15340172 TI - Improved stability of a protein vaccine through elimination of a partially unfolded state. AB - Ricin is a potent toxin presenting a threat as a biological weapon. The holotoxin consists of two disulfide-linked polypeptides: an enzymatically active A chain (RTA) and a galactose/N-acetylgalactosamine-binding B chain. Efforts to develop an inactivated version of the A chain as a vaccine have been hampered by limitations of stability and solubility. Previously, recombinant truncated versions of the 267-amino-acid A chain consisting of residues 1-33/44-198 or 1 198 were designed by protein engineering to overcome these limits and were shown to be effective and nontoxic as vaccines in mice. Herein we used CD, dynamic light scattering, fluorescence, and Fourier-transform infrared spectroscopy to examine the biophysical properties of these proteins. Although others have found that recombinant RTA (rRTA) adopts a partially unfolded, molten globule-like state at 45 degrees C, rRTA 1-33/44-198 and 1-198 are significantly more thermostable, remaining completely folded at temperatures up to 53 degrees C and 51 degrees C, respectively. Deleting both an exposed loop region (amino acids 34 43) and the C-terminal domain (199-267) contributed to increased thermostability. We found that chemically induced denaturation of rRTA, but not the truncated variants, proceeds through at least a three-state mechanism. The intermediate state in rRTA unfolding has a hydrophobic core accessible to ANS and an unfolded C-terminal domain. Removing the C-terminal domain changed the mechanism of rRTA unfolding, eliminating a tendency to adopt a partially unfolded state. Our results support the conclusion that these derivatives are superior candidates for development as vaccines against ricin and suggest an approach of reduction to minimum essential domains for design of more thermostable recombinant antigens. PMID- 15340173 TI - Sedimentation velocity studies of the high-molecular weight aggregates of the HIV gp41 ectodomain. AB - Accumulation of the HIV envelope protein gp41 is observed in the brain tissues of patients suffering from HIV-associated dementia. Previously, we have shown by electron microscopy that the extracellular domain of SIV gp41, which is analogous to that of HIV, forms high-molecular weight aggregates in vitro, and we have postulated that such high-molecular weight aggregates are responsible for neurological damage in a manner similar to protein deposition diseases such as Alzheimer's and the prion diseases. In this manuscript, we have characterized the self-association of the HIV gp41 ectodomain by sedimentation velocity. We show that discreet species of the gp41 high-molecular weight aggregates are present. The maximum population occurs at 20 S, which corresponds to ~5 trimers of gp41, suggesting that five trimers are required for nucleation of the high-molecular weight aggregates. The concentration dependence of the gp41 self-association indicates that it occurs by mass action. The temperature dependence of gp41 self association suggests that it is driven by entropy, indicating that intermolecular hydrophobic interactions between trimers of gp41 are driving the association. PMID- 15340174 TI - Circular permutation as a tool to reduce surface entropy triggers crystallization of the signal recognition particle receptor beta subunit. AB - The production of diffraction-quality crystals remains a difficult obstacle on the road to high-resolution structural characterization of proteins. This is primarily a result of the empirical nature of the process. Although crystallization is not predictable, factors inhibiting it are well established. First, crystal formation is always entropically unfavorable. Reducing the entropic cost of crystallizing a given protein is thus desirable. It is common practice to map boundaries and remove unstructured regions surrounding the folded protein domain. However, a problem arises when flexible regions are not at the boundaries but within a domain. Such regions cannot be deleted without adding new restraints to the domain. We encountered this problem during an attempt to crystallize the beta subunit of the eukaryotic signal recognition particle (SRbeta), bearing a long and flexible internal loop. Native SRbeta did not crystallize. However, after circularly permuting the protein by connecting the spatially close N and C termini with a short heptapeptide linker GGGSGGG and removing 26 highly flexible loop residues within the domain, we obtained diffraction-quality crystals. This protein-engineering method is simple and should be applicable to other proteins, especially because N and C termini of protein domains are often close in space. The success of this method profits from prior knowledge of the domain fold, which is becoming increasingly common in today's postgenomic era. PMID- 15340175 TI - Quantitative information management for the biochemical computation of cellular networks. AB - Understanding complex protein networks within cells requires the ability to develop quantitative models and to numerically compute the properties and behavior of the networks. To carry out such computational analysis, it is necessary to use modeling tools and information management systems (IMSs) where the quantitative data, associated to its biological context, can be stored, curated, and reliably retrieved. We have focused on the biochemical computation of cellular interactions and developed an IMS that stores both quantitative information on the cellular components and their interactions, and the basic reactions governing those interactions. This information can be used to construct pathways and eventually large-scale networks. This system, SigPath, is available on the Internet (http://www.sigpath.org). Key features of the approach include (i) the use of background information (for example, names of molecules, aliases, and accession codes) to ease data submission and link this quantitative database with other qualitative databases, (ii) a strategy to allow refinement of information over time by multiple users, (iii) the development of a data representation that stores both qualitative and quantitative information, and (iv) features to assist contributors and users in assembling custom quantitative models from the information stored in the IMS. Currently, models assembled in SigPath can be automatically exported to several computing environments, such as Kinetikit/Genesis, Virtual Cell, Jarnac/JDesigner, and JSim. We anticipate that, when appropriately populated, such a system will be useful for large-scale quantitative studies of cell-signaling networks and other cellular networks. SigPath is distributed under the GNU General Public License. PMID- 15340176 TI - Vascular remodeling in atherosclerotic femoral arteries: three-dimensional US analysis. AB - PURPOSE: To investigate the various modes of vascular remodeling of atherosclerotic femoral arteries and determine the associations between type of arterial remodeling and clinical data (age, sex, and medical history) and inflammatory parameters. MATERIALS AND METHODS: Intravascular ultrasonography (US) of the femoral arteries was performed in 50 patients with clinical symptoms of peripheral vascular disease. To determine the arterial remodeling mode (expansive remodeling [ER], involving compensatory enlargement of the artery, or constrictive remodeling [CR], involving vessel constriction during progression of atherosclerosis), the cross-sectional areas (CSAs) of the external elastic membrane (EEM), lumen, and plaque-plus-media were measured every 0.1 mm by using three-dimensional reconstruction. Clinical, laboratory, and intravascular US data were compared in the different remodeling groups (dominant ER, dominant CR, or mixed remodeling) by using analysis of variance supplemented by Tukey-Kramer tests. Multivariate analysis was performed to test independent variables predicting dominant ER. RESULTS: Intravascular US revealed the parallel existence of ER and CR in all patients: Increases and decreases in EEM in response to plaque growth could be observed within the same artery. ER dominated in 13 (26%) patients, and CR dominated in 11 (22%) patients: At least 80% of EEM CSAs were higher or lower than the mean of the EEM CSAs of the segments proximal and distal to the lesion. Patients with dominant ER had higher levels of serum C-reactive protein (CRP) compared with levels in patients with dominant CR and patients with mixed remodeling (1.62 mg/dL +/- 2.05 [standard deviation] vs 0.19 mg/dL +/- 0.33 and 0.21 mg/dL +/- 0.39, respectively, P < .005). Multivariate analysis revealed high CRP level to be a significant independent predictor for dominant ER (P < .01). CONCLUSION: The parallel existence of ER and CR was found in all patients with peripheral atherosclerosis, with a dominance of vessel expansion in 26% of patients. Higher plasma CRP level was associated with dominant ER. PMID- 15340177 TI - Acute stroke assessment with CT: do we need multimodal evaluation? AB - PURPOSE: To assess detection of stroke and prediction of extent of infarction with multimodal computed tomographic (CT) evaluation (unenhanced CT, perfusion CT, and CT angiography) in patients suspected of having acute stroke. MATERIALS AND METHODS: Forty-four consecutive patients with a mean National Institutes of Health Stroke Scale score of 10.45 and suspected of having ischemic stroke of the anterior circulation were examined with multi-detector row CT within 8 hours (mean, 3.05 hours) of onset of symptoms. All evaluations were performed with the knowledge that acute stroke was suspected but without detailed clinical information. The extent of ischemia or final infarction on the baseline unenhanced CT scan and follow-up images was assessed with the Alberta Stroke Program Early CT score. Different perfusion maps and follow-up images were assessed to determine the percentage of the ischemia-affected hemisphere. Each component, as well as the multimodal CT evaluation, was compared with follow-up unenhanced CT scans or magnetic resonance images after a mean time of 2.32 days. RESULTS: Multimodal CT revealed true-positive findings in 30 of 41 patients and true-negative findings in three, resulting in a sensitivity of 78.9%. Unenhanced CT, CT angiography, and perfusion CT showed sensitivities of 55.3%, 57.9%, and 76.3%, respectively. In eight patients, small infarctions (mean size, 1.47 cm) that were proved at follow-up were missed with all modalities at initial multimodal CT. With perfusion CT, four of these small infarctions were missed within the white matter of the section levels. Maps of cerebral blood flow showed the best correlation with the final size of infarction with an r(2) value of 0.71. CONCLUSION: The presented multimodal CT evaluation improves detection rate and prediction of the final size of infarction in comparison with unenhanced CT, CT angiography, and perfusion CT alone. PMID- 15340178 TI - Multiple Chromosome Abnormalities Following Bone Marrow Transplant for Chronic Myelogenous Leukemia. AB - A 44-year-old female was diagnosed in the chronic phase of chronic myelogenous leukemia (CML) and was confirmed to be Philadelphia chromosome positive by a bone marrow cytogenetic study. No additional cytogenetic abnormalities were found. The patient's cell counts were initially well controlled with hydrox-yurea. She then received an unrelated 6 of 6 HLA matched allo-geneic bone marrow transplant (BMT) from a male donor. The patient underwent myeloablative therapy with thiotepa and five fractions of total body radiation prior to the transplant. About four weeks after transplantation, the patient developed biopsy-proven graft-versus-host disease of the skin and GI tract. A blood sample was drawn at that time for cytogenetic analysis. Among 34 analyzed cells, 22 were normal male donor cells. The remaining 12 cells did not have the t(9;22), but had numerous structural abnormalities. While many cells were missing an X chromosome, other abnormalities, including deletions, rearrangements, dicentrics, acentric fragments, rings and marker chromosomes were non-clonal. No clinical evidence of progression from CML chronic phase was found, suggesting that the non-clonal abnor-malities were therapy related. PMID- 15340179 TI - [Essential trace metals and brain function]. AB - Trace metals such as zinc, manganese, and iron are necessary for the growth and function of the brain. The transport of trace metals into the brain is strictly regulated by the brain barrier system, i.e., the blood-brain and blood cerebrospinal fluid barriers. Trace metals usually serve the function of metalloproteins in neurons and glial cells, while a portion of trace metals exists in the presynaptic vesicles and may be released with neurotransmitters into the synaptic cleft. Zinc and manganese influence the concentration of neurotransmitters in the synaptic cleft, probably via the action against neurotransmitter receptors and transporters and ion channels. Zinc may be an inhibitory neuromodulator of glutamate release in the hippocampus, while neuromodulation by manganese might mean functional and toxic aspects in the synapse. Dietary zinc deficiency affects zinc homeostasis in the brain, followed by an enhanced susceptibility to the excitotoxicity of glutamate in the hippocampus. Transferrin may be involved in the physiological transport of iron and manganese into the brain and their utilization there. It is reported that the brain transferrin concentration is decreased in neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease and that brain iron metabolism is also altered. The homeostasis of trace metals in the brain is important for brain function and also for the prevention of brain diseases. PMID- 15340180 TI - Coloration of phenothiazines with metal-containing drugs. AB - We studied the color change of phenothiazines and metal-containing drugs after compound formation, followed by use of FT-Raman spectrocopy to observe any structural changes in them. When 6 phenothiazines (thioridazine hydrochloride, prochlorperazine maleate, levomepromazine maleate, chlorpromazine phenolphthalinate, fluphenazine maleate and perphenazine fendiate) formed compounds with natural aluminum silicate, the color change was accompanied by a shift of FT-Raman signals. These changes could be attributed to the structural changes of phenothiazines. This present observation can be then used in advance to avoid coloration of phenothiazines during preparative procedures with metal containing drugs such as antacids. PMID- 15340181 TI - Pharmacokinetic/pharmacodynamic analysis of neutrophil proliferation induced by rhG-CSF in patients receiving antineoplastic drugs. AB - In the present study, we analyzed the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on neutrophil counts in cancer patients undergoing chemotherapy using a previously developed pharmacokinetic/pharmacodynamic model.(7)) The time profiles of neutrophil counts in blood after repeated administration of rhG-CSF to lung cancer patients undergoing chemotherapy could be analyzed by this model by considering the inhibition of neutrophil production by antineoplastic drugs. Although deviation was observed between the predicted and observed neutrophil counts in ovarian cancer patients, it may be possible to use this model for determining a rational dosage regimen of rhG-CSF for patients undergoing chemotherapy. PMID- 15340182 TI - [Effects of the flavonoid fraction from Ginkgo biloba extract on the postprandial blood glucose elevation in rats]. AB - The present study investigated the effects of Ginkgo biloba extract and its flavonoid fractions on alpha-amylase and alpha-glucosidase activity in vitro. Ginkgo biloba extracts and their flavonoid fraction significantly inhibited alpha amylase and alpha-glucosidase activity in vitro. Furthermore, Ginkgo biloba extracts and their flavonoid fraction reduced the elevation of rat plasma glucose level after oral administration of various saccharinity agents. In addition, we examined the effects of the flavonoid fraction isolated from Ginkgo biloba extracts on the plasma glucose level in streptozotocin-induced diabetic rats. When flavonoid fractions were orally administered to the rats three times daily for 9 days, plasma glucose concentrations were decreased compared with those in the water treatment group. Furthermore, flavonoid fractions reduced the elevation of rat plasma glucose levels after oral administration of sucrose and glucose in streptozotocin-induced diabetic rats. PMID- 15340183 TI - [KiBank: a database for computer-aided drug design based on protein-chemical interaction analysis]. AB - KiBank is a database for computer-aided drug design and consists of binding affinities and chemical and target protein structures. Each chemical or protein structure with hydrogen atoms added was optimized by energy minimization and stored in PDB or MDL MOL file format, so that the structural data can be directly used for in silico binding studies. To describe the extent of inhibition, the inhibition constant (K(i)) value is used to simplify comparisons of strengths among chemical-protein bindings. As of April 2004, KiBank contained 142 proteins, over 5000 chemicals, and over 6000 binding affinity values that were published in peer-reviewed journals. The binding affinity values are currently mostly for membrane and nuclear receptors but are soon being expanded to other drug targets. KiBank is updated daily and can be accessed on the Web at http://kibank.iis.u tokyo.ac.jp/at no charge. PMID- 15340184 TI - [Physical properties and metal elutions of the thirteen molten slags produced from incinerated ash]. AB - In recent years, there have been many attempts to reuse molten slag. Molten slag is obtained through further treatment of incinerated ash produced in the disposal process of garbage or sewage sludge and is used in building materials, such as brick. To establish the safety of such slag, its physical properties and metal contents were investigated in this study. After examining the physical and chemical characteristics of 13 forms of slag made using different methods, no differences were observed and it was judged that reuse is possible. There was little elution of toxic metals in the elution test when water was used as a solvent. On the other hand, when acid and alkali were used, the elution of toxic metals tended to become greater. PMID- 15340185 TI - Effect of incadronate on corticosteroid-induced osteopenia in rats. AB - The effect of incadronate, a third-generation bisphosphonate, was evaluated in rats with corticosteroid-induced osteopenia. Male Wistar rats were treated with methylprednisolone acetate (1 mg/kg, s.c.) once daily, 3 days a week for 12 weeks. Other groups received simultaneous treatment with methylprednisolone acetate and incadronate (0.03, 0.3 or 3 mg/kg, p.o.); incadronate was given once daily, 6 days a week for 12 weeks. Bone mineral densities (BMDs) of the second lumbar (L2) vertebra as well as the ultimate compressive strength of the fifth lumbar (L5) vertebra decreased. Incadronate dose-dependently inhibited the loss of L2 BMDs and the decrease in strength of the L5 vertebrae. These results suggest that incadronate may be effective in treating osteopenia accompanying corticosteroid therapy. PMID- 15340186 TI - Evaluation of serum albumin utilization in inpatient at a private hospital in Bangkok. AB - Serum albumin is widely used for the treatment of hypovolemia, shock, hyperbilirubinemia or acute liver failure. However, serum albumin itself may have a significant effect on the action, elimination or distribution of other medications in blood which may affect the outcome of the treatment. Besides, it is also expensive and, most important, can be a carrier for many life-threatening diseases such as AIDS. The objective of this study is to evaluate the serum albumin utilization in inpatient at a private hospital in Bangkok, Thailand. Descriptive, retrospective study was designed to monitor the 25% w/v serum albumin orders over a four-month period. The background information, doses and rational of serum albumin orders, other medication orders, side effects, the interaction between serum albumin and other medications, the laboratory results and also clinical outcome of the patients were recorded. From the medication orders of 74 inpatients, there were 104 orders of serum albumin for 9 indications. The most common indications were hypoalbuminemia (48%) and cirrhosis (9%). From the total number of serum albumin orders, there were only 67 orders (64.4%) prescribed with rational dose and duration under appropriate indications. Totally about 35.6% of serum albumin orders were prescribed with improper indications such as edema, anemia or coronary artery bypass grafting. Out of 35.6%, 14% of the orders were prescribed to contraindicated patients. No side effect of serum albumin infusion was found during this study. The result indicates the over-utilization of serum albumin and specific guideline for using this agent should be set in every hospital. PMID- 15340187 TI - Enantioselective total syntheses of several bioactive natural products based on the development of practical asymmetric catalysis. AB - I present herewith enantioselective total syntheses of several bioactive natural products, such as (-)-strychnine, (+)-decursin, (-)-cryptocaryolone diacetate, ( )-fluoxetine, and aeruginosin 298-A, based on practical asymmetric catalyses (Michael reaction, epoxidation, and phase-transfer reaction) that I developed with co-workers in Prof. Shibasaki's group over the past 5 years. In the first part of this review, I discuss the great improvement of catalyst efficiency in an ALB-catalyzed asymmetric Michael reaction of malonate and application to the pre manufacturing scale (greater than kilogram scale) and enantioselective total synthesis of (-)-strychnine with the development of novel domino cyclization. To broaden the substrate generality of the Michael reaction, we developed a highly stable, storable, and reusable La-O-linked-BINOL complex. Further extension of the reaction using beta-keto ester as a Michael donor was achieved with the development of a La-NR-linked-BINOL complex, thereby improving indole alkaloid syntheses. In the second section, I discuss enantioselective total synthesis of (+)-decursin using catalytic asymmetric epoxidation. To achieve the synthesis, we developed a new La-BINOL-Ph(3)As = O (1:1:1) complex catalyst system, which has much higher reactivity and broader substrate generality than the previously developed catalyst systems. This allowed us to achieve catalytic asymmetric epoxidation of alpha,beta-unsaturated carboxylic acid derivatives with high enantioselectivity and broad substrate generality for the first time by changing the lanthanide metal and reaction conditions. Among them, catalytic asymmetric epoxidation of alpha,beta-unsaturated morpholinyl amides is quite useful in terms of synthetic utility of the corresponding alpha,beta-epoxy morpholinyl amides. Highly catalyst-controlled enantio- or diastereoselective epoxidation of the alpha,beta-unsaturated morpholinyl amides, coupled with diastereoselective reduction of beta-hydroxy ketones, enabled the synthesis of all possible stereoisomers of 1,3-polyol arrays with successful enantioselective total synthesis of several 1,3-polyol natural products, such as (-)-cryptocaryolone diacetate. In addition, the development of a new regioselective epoxide-opening reaction of alpha,beta-epoxy amides to the corresponding alpha- and beta-hydroxy amides enhanced the usefulness of the present epoxidation and was applied to the enantioselective total synthesis of (-)-fluoxetine. In the final section, I report the development of a new asymmetric two-center organocatalyst (TaDiAS) and its application to the enantioselective synthesis of aeruginosin 298-A and its analogues. Because of the remarkable structural diversity of TaDiAS, a practical asymmetric phase-transfer reaction with broad substrate generality was achieved. As a result, we succeeded in developing a highly versatile synthetic method for aeruginosin 298-A and its analogues. Inhibitory activity studies of the compounds against the serine protease trypsin provided preliminary information about their structure-activity relations. PMID- 15340188 TI - Spectroscopic studies on the mode of interaction of an anticancer drug with bovine serum albumin. AB - The mechanism of interaction of vinblastin sulphate (VBS) with bovine serum albumin (BSA) has been reported. Association constant for VBS-BSA binding was found to be 3.146+/-0.06 x 10(4) M(-1). Stern-Volmer analysis of fluorescence quenching data showed that the fraction of fluorophore (protein) accessible to the quencher (drug) was close to unity indicating thereby that both tryptophan residues of BSA are involved in drug-protein interaction. The rate constant for quenching, greater than 10(10) M(-1) S(-1), indicated that the drug-binding site is in close proximity to tryptophan residues of BSA. Binding studies in the presence of an hydrophobic probe, 8-anilino-1-naphthalein-sulphonic acid, sodium salt (ANS) indicated that there is hydrophobic interaction between VBS and probe and they do not share common sites in BSA. Thermodynamic parameters obtained from data at different temperatures showed that the binding of VBS to BSA involves predominant hydrophobic forces. The effects of some additives and paracetamol on binding of VBS-BSA have also been investigated. The CD spectrum of BSA in presence of VBS shows that the binding of VBS leads to change in the helicity of BSA. PMID- 15340189 TI - Size control of mixed egg yolk phosphatidylcholine (EggPC)/oleate vesicles. AB - The size distribution after addition of oleate surfactant to preformed Egg yolk phosphatidylcholine (EggPC) vesicles was investigated by gel filtration chromatography combining with dynamic light scattering. Phospholipid and oleate concentration, fluorescence intensity and size of the vesicles were measured for each elution fraction. The spontaneous vesiculation of oleate at pH 8.5 was accelerated in the presence of preformed EggPC vesicles. The size distribution of newly formed vesicles was dependent on the preformed vesicular size. For example, oleate addition to large preformed vesicles (230 nm) resulted in altering of vesicles to both larger and smaller than preformed ones, while addition of oleate to small preformed vesicles (50 nm) led to the formation of only larger vesicles without exhibition of newly small vesicles. The combinations of gel filtration chromatography and dynamic light scattering could provide more detailed insight into the size change of newly formed vesicles. PMID- 15340190 TI - Electrochemical study of zolpidem at glassy carbon electrode and its determination in a tablet dosage form by differential pulse voltammetry. AB - The oxidative behaviour of, a hypnotic drug, zolpidem was studied at glassy carbon electrode in Britton-Robinson buffer over the pH range 2.0-11.0 using cyclic, linear sweep and differential pulse voltammetry. Oxidation of the drug was effected in a single irreversible, diffusion-controlled step. Using differential pulse voltammetry (DPV), the drug yielded a well-defined voltammetric response in Britton-Robinson buffer, pH 8.0 at +0.889 V (vs. Ag/AgCl) on glassy carbon electrode. This process could be used to determine zolpidem concentrations in the range 5.0 x 10(-7) M to 1.0 x 10(-5) M with a detection limit of 2.0 x 10(-7) M. The method was applied, without any interference from the excipients, to the determination of the drug in a tablet dosage form. PMID- 15340191 TI - Improved dissolution of an insoluble drug using a 4-fluid nozzle spray-drying technique. AB - A solid dispersion of the drug can be made using a polymer carrier to improve solubility. Generally, drugs become amorphized when solid dispersion is formed using a polymer carrier. In such high energy conditions, the solubility of the drug molecule is increased. We previously prepared solid dispersion using a spray drying technique and reported its solubility and crystallinity. In this study, hydroxypropylmethylcellulose (HPMC) was used as the carrier, and tolubutamide was the model drug, which is water-insoluble. Solubility was evaluated by preparing a solid dispersion using a newly developed 4-fluid nozzle spray dryer. Observation of particle morphology by scanning electron microscopy (SEM) revealed that the particles from the spray drying were atomized to several microns, and they had also become spherical. Assessment of the crystallinity of the spray-dried particles by powder X-ray diffraction and differential scanning calorimetry demonstrated that the tolbutamide had been amorphized, forming a solid dispersion. The apparent release rate constant K of the drug from the spray-dried particles was 4 to 6 times faster than the original drug in pH 1.2, and it was also 1.5 to 1.9 times faster than the original drug in pH 6.8. The 70% release time (T(70)) of the drug from the spray-dried particles was 20 to 30 times faster than the original drug in pH 1.2 solution as well as 2 to 3 times faster than the original drug in pH 6.8 solution. Pharmaceutical preparations prepared in this way using the 4-fluid nozzle system spray dryer formed composite particles, resulting in a remarkably improved dissolution rates of the drug. PMID- 15340192 TI - Synthesis and evaluation of novel pyrimido-acridone, -phenoxadine, and -carbazole as topoisomerase II inhibitors. AB - As part of a series of studies to discover new topoisomerase II inhibitors, novel pyrimidoacridones, pyrimidophenoxadines, and pyrimidocarbazoles were synthesized, and in vitro and in vivo antitumor activities and DNA-protein and/or DNA topoisomerase II cross-linking activity as an indicator of topoisomerase II-DNA cleavable complex formation were evaluated. The pyrimidocarbazoles possessed high in vitro and in vivo potencies. Compound 26 (ER-37326), 8-acetyl-2-[2 (dimethylamino)ethyl]-1H-pyrimido[5,6,1-jk]carbazole-1,3(2H)-dione, showed in vitro growth inhibitory activity with respective IC(50) values of 0.049 microM and 0.35 microM against mouse leukemia P388 and human oral cancer KB. In vivo, this compound inhibited the tumor growth of mouse sarcoma M5076 implanted into mice with T/C values of 42% and 13% at 3.13 and 6.25 mg/kg/d respectively without significantly affecting the body weight. In addition, compound 26 (ER-37326) increased the formation of DNA-topoisomerase II cross-linking in P388 cells. PMID- 15340193 TI - A new synthetic method for an indolizidine skeleton by C-N bond formation via a pi-allylpalladium complex. AB - Pd(0)-catalyzed intramolecular cyclic reaction via a pi-allylpalladium complex provided an indolizidine skeleton in satisfactory high and reproducible yields by using allylic compound having an acetoxyl group as a leaving group. These results must be available for syntheses of various functional indolizidine alkaloids. PMID- 15340194 TI - Terpenic and phenolic glycosides from leaves of Breynia officinalis HEMSL. AB - From the leaves of Breynia officinalis, collected on Okinawa Island, six terpenic glucosides and six phenolic glycosides were isolated. Two of the terpenic glucosides were found to be known, and they were identified as turpinionoside B and betulalbuside A. The structures of the remaining terpenic glucosides were elucidated to be megastigmane glucosides, named breyniaionosides A-D, using spectroscopic analyses. The absolute structure of breyniaionoside D was determined using the modifed Mosher's method. The absolute structure of the known compound betulalbuside A was determined for the first time in this study. Six phenolic glycosides were found to be arbutin and its derivatives. Two known compounds were found to be robustaside A and eximine. New compounds were named isorobustaside A, and breyniosides A and B, and their structures were elucidated from spectroscopic evidence. PMID- 15340195 TI - Novel antiviral fucoidan from sporophyll of Undaria pinnatifida (Mekabu). AB - Structural characterization and antiviral activities of fucoidan from sporophyll of Undaria pinnatifida (Mekabu) was examined. The fucoidan was composed of fucose and galactose with an approximately ratio of 1.0:1.1. Degree of substitution of sulfate was 0.72 and its apparent molecular weight was 9,000. Methylation analyses showed that fucoidan had various sugar linkages, and revealed that the fucoidan might have complicated structure. This fucoidan showed potent antiviral activities against herpes simplex virus type 1 (HSV-1), HSV-2, and human cytomegalovirus. PMID- 15340196 TI - New products from the reaction of acetyllycoctonine with N-bromosuccinimide (NBS). AB - Six new artificial products 11-16 were obtained from the reaction of acetyllycoctonine (10) with N-bromosuccinimide (NBS). The structures of these compounds were established on the basis of spectral data. It is emphasized that the varieties and yields of the products in this reaction depended greatly upon reaction conditions and the types of substrate. PMID- 15340197 TI - Discovery of new orally active phosphodiesterase (PDE4) inhibitors. AB - A series of 4-anilinopyrazolopyridine derivatives were synthesized and biologically evaluated as inhibitors of phosphodiesterase (PDE4). Chemical modification of 3, a structurally new chemical lead that was found in our in house library, was focused on 1- and 3-substituents. Full details of the discovery of a new orally active chemical lead 5 are presented. Structure activity relationship data, pharmacological evaluation, and the subtype selectivity study are also presented. PMID- 15340198 TI - Chemoenzymatic synthesis of n-hexyl and O-beta-D-xylopyranosyl-(1-->6)-beta-D glucopyranosides. AB - Direct beta-glucosidation between 1,6-octanediol (5) and D-glucose (3) using the immobilized beta-glucosidase (EC 3.2.1.21) from almonds with the synthetic prepolymer ENTP-4000 gave a mono-beta-glucoside (6) in 61.4% yield, which was converted into the n-hexyl beta-D-glucopyranoside (1) by means of a chemoenzymatic method. The coupling of the n-hexyl beta-D-glucopyranoside congener (13) and 2,3,4-tri-O-acetyl-beta-D-xylosyl congener (14), followed by deprotection, afforded the synthetic n-hexyl O-beta-D-xylopyranosyl-(1-->6)-beta D-glucopyranoside (2), which was identical to the natural 2 with respect to the spectral data and specific rotation. PMID- 15340199 TI - Synthesis of nitrogen-functionalized cyclohexanes using chemoselective conjugate addition of phenyllithium to linear omega-nitro-alpha,beta,psi,omega-unsaturated ester and subsequent stereoselective intramolecular nitro-Michael cyclization. AB - Nitrogen-functionalized cyclohexane derivatives with three contiguous chiral centers were synthesized by nitroalkene-selective conjugate addition of phenyllithium to a omega-nitro-alpha,beta,psi,omega-unsaturated ester and subsequent stereocontrolled intramolecular nitro-Michael cyclization with cesium fluoride and a quaternary ammonium bromide. The cyclohexanes were applicable to the total synthesis of alpha-, beta- and gamma-lycoranes. PMID- 15340200 TI - Synthesis and Diels-Alder reactivity of ortho-carbazolequinones. AB - Oxidation of 2- and 3-hydroxycarbazoles with Fremy's salt gave the corresponding ortho-carbazolequinones. These molecules react as carbodienophiles in Diels-Alder reaction with 1-acetoxy-1,3-butadiene and 1,3-cyclopentadiene to provide the novel benzocarbazolequinone structures 15, 16, 18 and 19. PMID- 15340201 TI - Study on the stevioside analogues of steviolbioside, steviol, and isosteviol 19 alkyl amide dimers: synthesis and cytotoxic and antibacterial activity. AB - A new group of steviolbioside amide dimers 2a-g, derivatives 2h-i and their related steviol and isosteviol amide dimers 3a and 4a were prepared by reacting aliphatic alkylamine and alkyldiamines with PyBOP and DIEA. The synthesized compounds had cytotoxic effects on cancer and human embryonic lung cells. Compounds 3a, 4a, 2b and 2h were cytotoxic to cancer cells and to a lesser extent to human embryo lung cells. Compounds 2f, 2g and 4 of this series had favorable antibacterial effects, and were superior to penicillin G at inhibiting growth of Bacillus subtilis (BCRC 10029). The cytotoxicity and antibacterial effects may depend on the dimerization and derivative moieties in relation to the respective aglycons. PMID- 15340202 TI - Regioselective enzymatic acetylation of the aglycone moiety of a secoiridoid glucoside. Two new secoiridoid glucoside acetates. AB - Candida antarctica lipase (CAL) catalyses the regioselective acetylation of the 10-hydroxyl group of 10-hydroxyoleoside dimethyl ester, a secoiridoid glucoside, using THF as a solvent and ethyl acetate or vinyl acetate as acetyl group suppliers. Two acetyl derivatives at 3'- and 6'-sites of the glucosidic ring of 10-acetoxyoleoside dimethylester, not previously described, were obtained by acetylation in the same conditions. PMID- 15340203 TI - Examination of the reactivity of hydroxy groups in multioxygenated cyclohexanoids: synthetic study toward cytotoxic pericosine B. AB - The reactivity of two hydroxy groups in some multioxygenated cyclohexanoids was examined for basic study of the synthesis of the cytotoxic marine natural product pericosine B and related compounds. Differences in reactivity for O-methylation or O-acylation among substrates were observed. PMID- 15340204 TI - Synthesis of the epimer of pericosine B from (-)-quinic Acid. AB - Synthesis of the epimer of pericosine B from (-)-quinic acid was achieved. This synthesis involves some regioselective and stereoselective processes. The desired product showed lower cytotoxic activity in comparison with natural pericosine B against the P388 leukemia cell line. The result implies that the stereocenter of C-6 in natural pericosine B plays an important role in this activity. PMID- 15340205 TI - Sesquiterpene esters from the fruits of Celastrus orbiculatus. AB - Three new beta-dihydroagarofuran sesquiterpene esters, 6 alpha-acetoxy-9 beta benzoyloxy-1 beta-cinnamoyloxy-8 beta-butanoyloxy-beta-dihydroagarofuran (1), 6 alpha-acetoxy-9 beta-benzoyloxy-1 beta-cinnamoyloxy-8 beta-(2-methylbutanoyloxy) beta-dihydroagarofuran (2), and 6 alpha-acetoxy-1 beta,8 beta-dibenzoyloxy-9 beta hydroxy-beta-dihydroagarofuran (6), together with three known compounds (3-5) were isolated from the fruits of Celastrus orbiculatus THUNB. Their structures were elucidated on the basis of spectroscopic methods. Compound 4 showed moderate activity in inhibiting LPS-induced nitric oxide production in murine macrophage RAW264.7 cells with an IC(50) value of 43.7 microM. PMID- 15340206 TI - Improved preparation of racemic 2-amino-3-(heteroaryl)propanoic acids and related compounds containing a furan or thiophene nucleus. AB - Racemic 2-amino-3-(heteroaryl)propanoic acids (1), mostly with a furan or thiophene nucleus as a heteroaryl group, were synthesized in 48-94% yield by the reduction of 3-(heteroaryl)-2-(hydroxyimino)propanoic acids (5) with zinc dust and formic acid in the presence of a catalytic amount of iron dust at 60 degrees C for 2 h. Under these conditions, unfavorable hydrogenolysis of bromine on the thiophene nucleus does not occur. Traditional N-formylation of the prepared 3 (heteroaryl)alanine (1) with a mixture of formic acid and acetic anhydride afforded 2-(formylamino)-3-(heteroaryl)propanoic acids (6) in 51-95% yield. PMID- 15340207 TI - Constituents of Crinoidea. 3. Isolation and structure of a glycosyl inositolphosphoceramide-type ganglioside with neuritogenic activity from the feather star Comanthus japonica. AB - A glycosyl inositolphosphoceramide-type ganglioside, CJP4, was obtained from the polar lipid fraction of the chloroform/methanol extract of the feather star Comanthus japonica. The structure of this ganglioside has been determined on the basis of chemical and spectroscopic evidence to be 9-O-methyl-(N-glycolyl-alpha-D neuraminosyl)-(2-->11)-9-O-methyl-(N-glycolyl-alpha-D-neuraminosyl)-(2-->11)-9-O methyl-(N-glycolyl-alpha-D-neuraminosyl)-(2-->3)-inositolphosphoceramide, which contains C(16)-sphingosine and C(22:0)-, C(24:0)-fatty acid. This is the first report on the isolation and structural elucidation of a trisialo-glycosyl inositolphosphoceramide-type ganglioside. Moreover, CJP4 exhibited neuritogenic activity toward the rat pheochromocytoma PC12 cells in the presence of nerve growth factor. PMID- 15340208 TI - Oxidation of 3-hydroxypiperidines with iodosylbenzene in water: tandem oxidative Grob fragmentation-cyclization reaction. AB - Oxidation of 3-hydroxypiperidine with iodosylbenzene in water afforded 2 pyrrolidinone directly in good yields. The reaction probably involves oxidative Grob fragmentation yielding imino aldehyde, which upon hydrolysis produces 2 pyrrolidinone via a cyclization-oxidation sequence. PMID- 15340209 TI - Two new analogues of trijugin-type limonoids from the leaves of Sandoricum koetjape. AB - Two new additional trijugin-type limonoids, sandrapins D (4) and E (5), which are analogues of the previously reported sandrapins A-C (1-3), were isolated as minor components from the leaves of Sandoricum koetjape (Meliaceae), and their structures were elucidated on the basis of MS and NMR spectral data. PMID- 15340210 TI - Two cangorosin A type triterpene dimers from Maytenus chuchuhuasca. AB - Two new cangorosin A type triterpene dimers, which composed of two triterpene units jointed by two ether linkages between the A and B rings, were isolated from the Brazilian medicinal plant "xuxua" (Maytenus chuchuhuasca). Structures of new isolates, xuxuasins A (1) and B (2), were established based on several spectroscopic evidences. PMID- 15340211 TI - A novel phenylpropenoyl sulfonic acid and a new chlorophyll from the leaves of Petasites formosanus KITAMURA. AB - A novel phenylpropenoyl sulfonic acid, petasiformin-A (1), and a new chlorophyll, petasiphyll-A (2), were isolated from the leaves of Petasites formosanus KITAMURA (Compositae). Their structures were established by spectral and chemical transformation methods. Petasiformin-A (1) showed the significant antioxidative activity in DPPH radical scavenging assay. PMID- 15340212 TI - Synthesis of optically active methyl 7 beta-hydroxykaurenoate with potent neuroprotective activity. AB - (-)-Methyl 7 beta-hydroxykaurenoate (3) and its 4-demethyl acetate (-)-4 were both synthesized via methods that contained radical cyclization and intramolecular Diels-Alder reactions as key steps. Both compounds displayed potent neuroprotective activity against N-methyl-D-aspartate toxicity in cultured cortical neurons. PMID- 15340213 TI - Antimicrobial activity of saturated fatty acids and fatty amines against methicillin-resistant Staphylococcus aureus. AB - The objective of this study was to investigate the antimicrobial activities of saturated fatty acids and fatty amines against methicillin-resistant Staphylococcus aureus (MRSA). The antimicrobial activity of saturated fatty acids and fatty amines was determined by oxygen meters with multi-channels and disposable oxygen electrode sensors (DOX-96). Lauric acid, the most effective among the saturated fatty acids, showed antimicrobial activity at 400 microg/ml against methicillin-susceptible Staphylococcus aureus (MSSA) and MRSA. The minimal inhibitory concentration (MIC) of fatty amines depended on each hydrophobic chain length. The MIC of myristylamine was 1.56 microg/ml; most effective of the fatty amines. In time-kill curves, lauric acid and myristylamine produced a bactericidal effect and a bacteriostatic effect at 4-fold the MIC, respectively. The antimicrobial activities of lauric acid and myristylamine were decreased by human plasma. Cytotoxicity of 3 saturated fatty acids and 3 fatty amines was examined in cultured endothelial cells. Although cytotoxicity of fatty amines was severer than that of saturated fatty acids, myristylamine showed the highest value of apparent therapeutic index among them. DOX-96 was useful for screening antimicrobial substances, especially in the case of insoluble substances. We found that myristylamine showed anti-MRSA activity comparable to that of vancomycin and teicoplanin. PMID- 15340214 TI - Mammalian spermidine synthase--identification of cysteine residues and investigation of the putrescine binding site--. AB - Homology modeling and inhibitory studies using substrate analogs were undertaken to construct a possible three-dimensional structure, including the putrescine binding site, of rat spermidine synthase based on its primary sequence. Of the ten cysteine residues of the enzyme, six residues were chemically determined as sulfhydryl; similarly, one residue (C25) was determined as the disulfide. Using the model obtained from the Swiss-Model protein-modeling server, and based on the crystal structure of the Thermotoga maritima enzyme, the three remaining residues were assigned as sulfhydryl. Discussions are presented on the counterpart of the C25 residue, based on the apparent role of the bacterial N-terminal peptide region in reinforcing the binding between protomers in a functional oligomeric form. The active sites of the bacterial and mammalian versions of the enzyme were very similar. The putrescine-binding site of the rat enzyme was investigated using IC(50) values of the analogs of two known potent inhibitors, n-butylamine and trans-4-methylcyclohexylamine (4MCHA). Our results indicated that 5-amino-1 pentene and 4MCHA possess comparable inhibitory activities towards the enzyme. PMID- 15340215 TI - New qualitative detection methods of genetically modified potatoes. AB - In Japan, 8 lines of genetically modified (GM) potato (2 lines of NewLeaf potato; NL, 3 lines of NewLeaf Plus potato; NLP, and 3 lines of NewLeaf Y potato; NLY) have already been authorized as safe for use in foods and feeds. We have developed polymerase chain reaction (PCR) methods for the qualitative detection of the GM potatoes for the screening and the identification of NL, NLP and NLY. The gene encoding uridine diphosphate (UDP)-glucose pyrophosphorylase (UGPase) was used as a taxon specific gene. We designed the primer pair to detect the cryIIIA genes as a screening method for GM potatoes because the gene should be inserted in all 8 lines of the GM potatoes. For identification of NL, NLP and NLY, we further designed three specific primer pairs for the different recombinant DNAs (r-DNA) specifically introduced into NL, NLP, or NLY. In addition, to identify the 3 lines of NLY that have been introduced with the same r-DNA, the three line-specific primer pairs for the border sequence between the r DNA and genomic DNA of NLY 3 lines were designed. Six lines of GM potato used as the test material were specifically identified using the each primer pair under the same PCR condition. The detection limits of all the GM potatoes should be approximately 0.1%. Furthermore, the specificity and reproducibility of the methods were confirmed in a six-laboratory collaborative study. PMID- 15340216 TI - Over-expressed Bcl-2 cannot suppress apoptosis via the mitochondria in buprenorphine hydrochloride-treated NG108-15 cells. AB - We previously reported that the morphine alkaloid derivative buprenorphine hydrochloride (Bph) induces rapid apoptosis in NG108-15 nerve cells accompanied by the activation of caspase-3. Here, we found this kind of apoptosis was also accompanied by rapid loss of the mitochondrial membrane potential, followed by the efflux of cytochrome c from the mitochondria to the cytosol and the activation of caspases-9 and -3. Together, these results strongly suggested the Bph death signal was routed through the mitochondrial pathway in NG108-15 cells. In these cells, serum-starvation induces a different apoptosis, which we exploited to investigate Bcl-2's role as an apoptosis inhibitor. We made an NG108 15 transfectant, Bcl-2(P2), that stably expressed human Bcl-2, and used it to test Bcl-2's effect on the serum-starvation-induced apoptosis in NG108-15 cells. Cell viability, DNA-ladder formation, and efflux of cytochrome c from the mitochondria were all detected, showing that the human Bcl-2 functioned normally in the Bcl-2(P2) cells. Although the apoptotic events tested were identical in the parental cells and transformants, Bcl-2 expression completely failed to inhibit Bph-induced apoptosis in the Bcl-2(P2) cells. PMID- 15340217 TI - Methylene chloride fraction of Spatholobi Caulis induces apoptosis via caspase dependent pathway in U937 cells. AB - Spatholobi Caulis has been used in Oriental medicine to treat cancer and blood stasis. In this study, the methylene chloride fraction of Spatholobi Caulis (MCSC) was examined to determine if it possesses anti-cancer activity via its apoptosis-inducing activity. MCSC exhibited a strong cytotoxic effect against human monocyte leukemia U937 cells (IC(50)=15.1 microg/ml). A TUNEL assay showed that the MCSC caused a characteristic ladder pattern of discontinuous DNA fragments and apoptotic bodies. Flow cytometric analysis confirmed that MCSC significantly increases the number of apoptotic cells stained by annexin V(+)/PI( ) cells. Western blotting revealed that MCSC activated caspase-3 expression and cleaved poly (ADP-ribose) polymerase (PARP) in a concentration-dependent manner. An enzyme-linked immunosorbent assay (ELISA) demonstrated that MCSC significantly activated the caspase-3 activity compared with the untreated control by. Taken together, these results suggest that MCSC can induce apoptosis in U937cells via the caspase dependent pathway. PMID- 15340218 TI - Effect of various catechins on the IL-12p40 production by murine peritoneal macrophages and a macrophage cell line, J774.1. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine comprising p40 and p35 subunits produced mainly by monocytes and macrophages, and plays an essential role in the regulation of the differentiation of Th1 cells. Green tea polyphenols exhibit potent anti-oxidative activities and anti-inflammatory effects by modulating cytokine production. We investigated the effect of catechins on IL 12p40 production in murine macrophages induced by bacterial lipopolysaccharide (LPS). Pretreatment with several catechins at doses of 0.3-30 microM suppressed IL-12 p40 production by murine peritoneal exudate cells (PEC) and J774.1 cells in a dose-dependent manner. Decreases in protein production were primarily due to down-regulation of the transcription of IL-12p40 mRNA. Of the various catechins, (-)-epigallocatechin gallate (EGCG) was the most potent inhibitor, followed by ( )-gallocatechin gallate (GCG) and (-)-epicatechin gallate (ECG). EGCG inhibited LPS-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), but not Jun N-terminal kinase (JNK), while EGCG augmented LPS-induced phosphorylation of p44/p42 extracellular signal-related kinase (ERK). In addition, both EGCG and GCG inhibited LPS-induced degradation of IkappaBalpha with concomitant inhibition of nuclear protein binding to NF-kappaB site and synthesis of IRF-1. These results suggest that gallate-containing catechins, particularly EGCG, inhibits LPS-induced IL-12p40 production in murine macrophages by inhibiting p38 MAPK while enhancing p44/p42 ERK, leading to the inhibition of IkappaBalpha degradation and NF-kappaB activation. PMID- 15340219 TI - Rubus croceacanthus Leveille inhibits mast cell-mediated anaphylactic-like reaction and tumor necrosis factor-alpha secretion. AB - This work aims at examining the effect of the concentrated methanol extract of Rubus croceacanthus Leveille (RCL) on mast cell-mediated anaphylactic-like reaction in a murine model. RCL inhibited compound 48/80-induced systemic anaphylactic-like reaction. When RCL was given as pre-treatment at concentrations ranging from 0.01 to 1 mg/ml, the histamine release from rat peritoneal mast cells induced by compound 48/80 or anti-dinitrophenyl (DNP) immunoglobulin E (IgE) was reduced in a dose-dependent manner. RCL also inhibited passive cutaneous anaphylaxis activated by anti-DNP IgE. In addition, RCL inhibited phorbol 12-myristate 13-acetate and A23187-induced tumor necrosis factor-alpha secretion from human mast cell line HMC-1 cells. These results indicate that RCL may possess a strong anti-anaphylactic activity. PMID- 15340220 TI - Effects of novel synthesized pyridothiazines on various guinea pig heart muscle preparations. AB - Calcium channel blockers have become important tools in the treatment of cardiovascular disorders and other diseases. Hybridization of well established calcium antagonist subclasses was an attempt to optimize their pharmacological profile. The intension of this study was to investigate the electrophysiological properties of MM 10 and MM 11 two newly synthesized compounds structurally closely related to KT-362 (5-[3-[[2-(3,4-dimethoxyphenyl)ethyl]amino]-1 oxopropyl]-2,3,4,5-tetrahydro-1,5-benzothiazepine fumarate) in various isolated guinea pig heart muscle preparations by means of the conventional intracellular microelectrode tech-nique. MM 10 (2,3-dihydro-1-[N-[2-(3,4-dimethoxyphenyl)ethyl] N-methylaminoacetyl]-1H-pyrido[2,3-b][1,4]thiazine fumarate) and MM 11 (2,3 dihydro-1-[N-[2-(3,4-dimethoxyphenyl)ethyl]-N-methylaminopropionyl]-1H-pyrido[2,3 b][1,4]thiazine fumarate) exerted very similar effects though the action of MM 11 was more pronounced. Whereas action potential amplitude and maximum upstroke velocity (V(max)) in papillary muscle, left atria and spontaneously beating Purkinje fibers was not affected by the compounds in a concentration range from 3 to 30 micromol/l, action potential duration at 90% time to repolarization was significantly prolonged in a concentration-dependent manner. Action potential duration at 20% time to repolarization was decreased in spontaneously beating Purkinje fibers and remained unchanged in papillary muscles and left atria. In sinoatrial nodes both compounds reduced rate of activity, action potential amplitude, maximum upstroke velocity and slope of slow diastolic depolarization while time to repolarization was prolonged. In 3 out of 6 experiments with spontaneously beating Purkinje fibers, MM 11 (30 micromol/l) led to the occurrence of early afterdepolarizations with a take off potential between -50 and -60 mV. All observed effects were completely reversible during washout with drug-free physiological salt solution. From these results it was concluded that both compounds in addition to their calcium antagonistic properties might depress repolarizing potassium currents. In contrast to the mother compound KT-362 they do not seem to affect the fast sodium inward current. Replacement of the benzothiazepine nucleus by a pyridothiazine structure may weaken or even eliminate sodium channel blocking ability. Shortening of the side chain might result in a general loss in activity. PMID- 15340221 TI - Effect of iron lactate overloading on adenine nucleotide levels and adenosine 3' monophosphate forming enzyme in rat liver and spleen. AB - To elucidate the pathophysiological significance of adenosine 3'-monophosphate (3'-AMP) forming enzyme in rats, the effect of iron lactate overloading on the enzyme activities and adenine nucleotide levels in the liver and spleen was examined. Sprague-Dawley rats were fed a diet supplemented with 0%, 0.625% or 5.0% of iron lactate for 4 weeks. Iron deposition was found in periportal hepatocytes, Kupffer cells and macrophages of red pulp of the spleen. No significant changes in hematological parameters were detected. Although serum alkaline phosphatase and inorganic phosphorus levels elevated slightly in the 5.0% group, activities of alanine aminotransferase and aspartate aminotransferase, and levels of serum urea nitrogen and creatinine were not changed significantly. The ATP levels in the liver and spleen of iron fed groups were significantly decreased, but adenosine 5'-diphosphate (ADP) and adenosine 5' monophosphate (AMP) levels were within control levels. On the other hand, the levels of ATP, ADP and AMP in the erythrocytes without mitochondria were not suppressed by the iron lactate overloading. Free activity of 3'-AMP forming enzyme, one of ribonucleases (RNase), was not changed in the liver of iron overloaded rat, and total amount of 3'-AMP and adenosine formed after the treatment of the crude enzyme(s) with p-chloromercuribenzensulfonic acid, a SH blocker of RNase inhibitors, was decreased dose-dependently. On the contrary, free activity of 3'-AMP forming enzyme was enhanced dose-dependently in the spleen of iron-overloaded rat but the total activity was not changed. However, the free and total 3'-AMP forming enzyme activities in the liver and spleen of iron-overloaded rats became equal at the dosage of 5.0% of iron lactate. The results obtained suggested that iron loading might induce significant decrease in hepatic and splenic ATP levels via malfunction of their mitochondria and might lead dissociation of RNase-RNase inhibitor complex to activate 3'-AMP forming enzyme in both tissues. PMID- 15340222 TI - Establishment of allergic dermatitis in NC/Nga mice as a model for severe atopic dermatitis. AB - Mite antigen has been suggested to play important roles in the onset and/or development of atopic dermatitis, and mite antigen-induced dermatitis models appear beneficial for the basic study of atopic dermatitis. In the present study therefore, we attempted to establish an allergic dermatitis model in mice using Dermatophagoides farinae crude extract as an antigen. Mite antigen solution at a concentration of 1 or 10 mg/ml was painted 5 times repeatedly at an interval of 7 d onto the ear of NC/Nga or BALB/c mice with or without simultaneous tape stripping. Apparent biphasic ear swelling was observed after the 4th and 5th antigen applications in both strains of mice treated with 10 mg/ml of antigen solution. Thickening of the epidermis, fibrosis of the dermis, and the accumulation of inflammatory cells were also observed after the 5th application. The inflammatory changes were more evident in NC/Nga mice than in BALB/c mice and potentiated by tape-stripping. The ear swelling was accompanied by increased serum IgE, increased expression of interleukin-4 mRNA and decreased expression of interferon-gamma mRNA in cervical lymph nodes and ears. These results indicate that ear swelling caused by repeated mite antigen application with simultaneous tape-stripping has a Th2-dominant background and that the inflammatory responses are expressed more potently in NC/Nga mice than in BALB/c mice. The dermatitis caused by mite antigen in NC/Nga mice appears to be a useful model for the basic study of atopic dermatitis. PMID- 15340223 TI - N(epsilon)-(carboxymethyl)lysine proliferated CD34(+) cells from rat choroidal explant in culture. AB - Action of N(epsilon)-(carboxymethyl)lysine-human serum albumin (CML-HSA) on neovascularization was investigated in cultured rat choroidal explant. Choroidal explants of normal male Wistar rats were cultured in fibrin gel with Dulbecco's modified Eagle medium containing fetal bovine serum in the presence or absence of CML-HSA. Migrated cells were budded from 2nd day in culture and developed from cultured choroidal explants in a time-dependent manner. Budded and developed cells from the choroidal explant had a feature of fibroblasts, which had attenuated long cytoplasmic processes, long ellipsoid nuclei and numerous membrane-bound polymorphic vesicles. Immunostaining of the attenuated cells in fibrin bed with CD34 (a marker protein of vascular endothelial cells and endothelial progenitor cells) failed to disclose positive result. However the cells which were isolated from fibrin bed by collagenase were specifically stained with anti-CD34 antibody. The isolated cells did not form tube-like structures on collagen gel by 3 weeks in culture. CML-HSA significantly increased the number of total isolated cells and CD34(+) cells as well as the number of vessel-like structures. These results indicate that CML-HSA overproduced immature blood vessels from cultured choroidal explants in fibrin gel, which consisted of CD34(+) cells. The CML-HSA-induced formation of immature blood vessel may be implicated in various choroidal diseases such as age-related macular degeneration. PMID- 15340224 TI - Neuroprotective effects of a dual L/N-type Ca(2+) channel blocker cilnidipine in the rat focal brain ischemia model. AB - Although a blockade or lack of N-type Ca(2+) channels has been reported to suppress neuronal pathological processes in several animal models of pain and ischemic brain injury, information is still limited regarding the neuroprotective effects of a dual L/N-type Ca(2+) channel blocker, cilnidipine. In this study, we assessed the effects of cilnidipine in the rat focal brain ischemia model to analyze its potential utility for hypertensive patients with cerebral infarction. In an anesthetized rat model, cerebral vasodilator actions of cilnidipine were detected at hypotensive doses, which was less potent than those of an L-type Ca(2+) channel blocker, nilvadipine. In the rat focal brain ischemia model, an anti-hypertensive and anti-sympathetic dose of cilnidipine could reduce the size of cerebral infarction, whereas an equipotent hypotensive dose of nilvadipine failed to affect it. These results suggest that N-type Ca(2+) channel-blocking profile of cilnidipine may contribute its neuroprotective action in the animal focal brain ischemia model. Thus, treatment of hypertension with cilnidipine may prevent severe consequences after brain attack. PMID- 15340225 TI - A novel immunomodulator, FTY720, prevents spontaneous dermatitis in NC/Nga mice. AB - Oral administration of a novel immunomodulator, FTY720 (0.1 mg/kg, once a week), completely prevented the spontaneous development of dermatitis in NC/Nga mice. It also strongly suppressed hyper IgE production in serum, as well as hypertrophy of the epidermis and the degranulation of granulocytes in the skin, all of which were observed in mice in which the dermatitis had become established. These results strongly suggest that FTY720 is a promising candidate for treatment of human atopic dermatitis. PMID- 15340226 TI - Reduction of the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice using an antiulcer drug, geranylgeranylacetone. AB - The protective effect of geranylgeranylacetone (GGA), an antiulcer drug, against the acute toxicity and teratogenicity produced by 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) was examined in C57BL/6J mice. When mice were co-treated, GGA reduced the loss of body weight gain and lethality produced by TCDD but hepatomegaly and thymic atrophy were not improved. Additionally, no protective effect of GGA was observed in the formation of cleft palate and hydronephrosis in mouse fetuses caused by maternal exposure to TCDD. To clarify the reducing mechanism by GGA, the Hsp70.1 mRNA levels in liver and intestine were analyzed. However, it was difficult to explain the effect of GGA from the induction of Hsp70.1. GGA had also no effect on the induction of hepatic ethoxyresorufin O deethylase activity by TCDD. These data suggest that GGA exhibits a protective effect against some forms of dioxin toxicity by a mechanism without involving inhibition of arylhydrocarbon receptor activation. PMID- 15340227 TI - Pharmacophore based synthesis of 3-chloroquinoxaline-2-carboxamides as serotonin3 (5-HT3) receptor antagonist. AB - A series of 3-chloroquinoxaline-2-carboxamides were designed and prepared by the condensation of 3-chloro-2-quinoxaloylchloride with appropriate Mannich bases of the p-aminophenol in the microwave environment. The synthesized compounds were evaluated for serotonin(3) (5-HT(3)) receptor antagonistic activities in longitudinal muscle-myenteric plexus (LMMP) preparation from guinea pig ileum against the 5-HT(3) agonist, 2-methyl-5-HT. Compound 3g exhibited comparable 5 HT(3) antagonistic activity (pA(2) 6.4) to that of standard antagonist Ondansetron (pA(2) 6.9), while the other compounds exhibited mild to moderate 5 HT(3) antagonistic activities. PMID- 15340228 TI - Suppressive effect of Kanzo-bushi-to, a Kampo medicine, on collagen-induced arthritis. AB - Kanzo-bushi-to (KBT) is a traditional Japanese herbal medicine (Kampo medicine), which is used in Japan to treat rheumatoid arthritis. In the present study, we investigated the suppressive effect of KBT on collagen-induced arthritis (CIA) and further studied the underlying mechanism. CIA was induced in male DBA/1J mice by immunization with bovine type II collagen, followed by a booster injection 21 d later. KBT was given at a dose of 430 mg/kg/d from three days before the first immunization to the end of the experiment. KBT suppressed CIA development effectively and further protected focal bone erosion and bone destruction as evidenced by the reduced histological score. Histochemical examination revealed that KBT decreased TRAP-positive cells at the synovium-bone interface and at the sites of focal bone erosion, coincident with the findings that RANKL/OPG mRNA ratio was significantly reduced by KBT treatment. KBT also decreased mRNA levels of M-CSF and iNOS in joints and of iNOS in peritoneal macrophages. In conclusion, KBT prevented osteoclast generation by decreasing RANKL/OPG ratio and M-CSF mRNA levels, resulting in reduction in bone erosion and destruction. In addition, KBT has anti-inflammatory effect such as the suppression of iNOS expression in peritoneal macrophages and joints of CIA mice. These finding suggests that KBT is a potential new therapeutic agent for the treatment of RA. PMID- 15340229 TI - Xanthine oxidase inhibitory activity of Vietnamese medicinal plants. AB - Among 288 extracts, prepared from 96 medicinal plants used in Vietnamese traditional medicine to treat gout and related symptoms, 188 demonstrated xanthine oxidase (XO) inhibitory activity at 100 microg/ml, with 46 having greater than 50% inhibition. At 50 microg/ml, 168 of the extracts were active, with 21 possessing more than 50% inhibition. At 25 microg/ml, 146 extracts exhibited inhibitory activity, with 8 showing over 50% inhibition, while 126 extracts presented activity at 10 microg/ml, with 2 having greater than 50% inhibition. The MeOH extracts of Artemisia vulgaris, Caesalpinia sappan (collected at the Seven-Mountain area), Blumea balsamifera (collected in Lam Dong province), Chrysanthemum sinense and MeOH-H(2)O extract of Tetracera scandens (Khanh Hoa province) exhibited strong XO inhibitory activity with IC(50) values less than 20 microg/ml. The most active extract was the MeOH extract of the flower of C. sinense with an IC(50) value of 5.1 microg/ml. Activity-guided fractionation of the MeOH extract led to the isolation of caffeic acid (1), luteolin (2), eriodictyol (3), and 1,5-di-O-caffeoylquinic acid (4). All these compounds showed significant XO inhibitory activity in a concentration-dependent manner, and the activity of 2 was more potent (IC(50) 1.3 microM) than the clinically used drug, allopurinol (IC(50) 2.5 microM). PMID- 15340230 TI - Intestinal absorption and hepatic extraction of propranolol and metoprolol in rats with bilateral ureteral ligation. AB - To investigate the mechanism responsible for the increased bioavailability of propranolol in bilateral ureter-ligated (BUL) rats, the intestinal absorption and hepatic extraction of propranolol and metoprolol were evaluated. The initial absorption rate of these drugs after intra-intestinal administration was only slightly increased in the BUL rats, whereas the blood drug concentration in these rats was higher than that in control rats. The blood propranolol and metoprolol concentrations during intra-portal infusion in the BUL rat were significantly higher than that in the control rat. In the presence of NADPH, the intrinsic metabolic activity of metoprolol in hepatic microsomes was not altered by BUL. On the other hand, the NADPH generation rate in the hepatic cytosol in the BUL group was lower than that in the control group. These results indicate that the absorption rate-dependent decrease in hepatic first-pass clearance of propranolol and metoprolol due to saturation kinetics is marginal, and that the hepatic metabolic activity and extraction of the drugs is significantly decreased in BUL rats probably due to the reduced NADPH generation rate in the liver. PMID- 15340231 TI - Difference in percutaneous absorption and intracutaneous distribution in guinea pigs among topical antifungal drugs (tioconazole solution, tioconazole cream, miconazole nitrate solution and bifonazole solution). AB - Tioconazole (TCZ) is an imidazole antifungal agent with broad spectrum activity. Percutaneous absorption and intracutaneous distribution of TCZ solution have been compared with TCZ cream, miconazole nitrate (MCZ) solution and bifonazole (BFZ) solution following a single topical application to abdominal skin of guinea pigs. Following application of TCZ solution, TCZ concentrations in the stratum corneum, epidermis-cutis and subcutaneous tissue were higher than those after TCZ cream application suggesting superior percutaneous penetration after TCZ solution application. The percutaneous penetration after applications of MCZ solution and BFZ solution was comparable to that of TCZ cream, but inferior to that of TCZ solution. TCZ concentrations in the stratum corneum were much higher than those in epidermis-cutis and subcutaneous tissue after applications of both TCZ formulations. The majority of applied TCZ remained in the stratum corneum at high levels for a long duration. TCZ concentrations in the stratum corneum within 24 h after applications of both TCZ formulations were more than several hundred times higher than the minimum inhibitory concentrations against most of the dermatophytes and yeasts. The effectiveness of both TCZ formulations against dermatophytoses may be due to this favorable pharmacokinetic property in the skin tissues, together with its potent antifungal activity. Percutaneous absorption of TCZ after applications of both formulations was negligible suggesting that these treatments are unlikely to produce systemic side effects. PMID- 15340232 TI - Preparation of translucent and flexible human hair protein films and their properties. AB - We have developed novel procedures for preparing human hair protein films (Pre cast and Post-cast methods). The light brown films obtained by these procedures were too fragile to apply to human skin. We found that the film was also formed when the hair proteins extracted by the Shindai method were directly exposed to the solution containing MgCl(2), CaCl(2), NaCl or KCl. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that the surface of the novel protein films was smooth. The protein films mainly consist of alpha keratins and matrix proteins. After drying, the films became translucent and flexible during folding, indicating the possibility that these protein films are useful for practical applications. Hence, we prepared gauze-coated protein films to reinforce their physical strength and tested the influence on human skin. A patch test showed that the protein films made from individual and multiple human hairs only slightly stimulated rubor and anthema, itching, drying, smarting and pain on the contact area of arm skin. PMID- 15340233 TI - mRNA expression of the ATP-binding cassette transporter subfamily A (ABCA) in rat and human brain capillary endothelial cells. AB - The ATP-binding cassette transporter subfamily A (ABCA) consists of the transporters mediating cholesterol release and regulated by cholesterol. As about 25% of total body cholesterol exists in the brain, sterol homeostasis is an important issue as far as central nervous system function is concerned. The purpose of this study was to clarify the mRNA expression of ABCA subtypes at the blood-brain barrier (BBB) using cultured rat and human brain capillary endothelial cells, TR-BBB and hBME cells, respectively. mRNA expression of ABCA1, 2, 3, 4, 5, 6, 7 and 8/9 was detected in TR-BBB cells. In the brain capillary rich fraction, mRNA expression of ABCA1, 2, 3, 4, 5, 7 and 8/9 was detected. ABCA2 and 5 mRNA were also detected in hBME cells. These results demonstrate, for the first time, that ABCA subtypes are expressed at the rat and/or human BBB. The expression of ABCA subtypes at the BBB is likely to contribute to sterol homeostasis in the central nervous system. PMID- 15340234 TI - Exogenous expression of interferon-beta in cultured brain microvessel endothelial cells. AB - Brain microvessel endothelial cells (BMECs) make up the blood-brain barrier (BBB) and regulate the passage of therapeutic proteins as well as drugs from the cerebrovasucular circulation to the brain. In the present study, we transferred mouse or human interferon-beta (IFN-beta) gene via cationic liposomes into primary cultures of bovine BMECs developed as an in vitro model of the BBB. The gene-transferred BMECs secreted transiently a substantial amount of IFN activity more efficiently during the growth phase than at confluence. This was suggested to be due to a difference in the potential for plasmid incorporation between growing and confluent BMECs in a series of cell association experiments with (32)P-labelled plasmid DNA. Furthermore, when BMEC monolayers in Transwell plates were transfected with the IFN-beta-expression vectors from the upper side, IFN beta was predominantly detected in the upper compartments, suggesting polarized secretion of the transgene products in BMEC monolayers. These findings provide important basic information about therapeutic secretory protein gene delivery to BMECs. PMID- 15340235 TI - Regucalcin increases superoxide dismutase activity in rat liver cytosol. AB - The effect of regucalcin, a regulatory protein in the intracellular signaling process, on superoxide dismutase (SOD) activity in the cytosol of rat liver was investigated. The presence of zinc sulfate (10(-6) or 10(-5) m) or cupric sulfate (10(-6) m) in the enzyme reaction mixture caused a significant increase in SOD activity, indicating that Cu/Zn-SOD may be present in the liver cytosol. SOD activity was significantly increased by the addition of regucalcin (0.1, 0.25, or 0.5 microM) to the reaction mixture. The presence of dithiothreitol (DTT; 0.1, 0.5, or 1.0 mM), a protective reagent for the sulfhydryl group, caused a significant decrease in SOD activity. The effect of regucalcin (0.25 microM) in increasing SOD activity was not seen in the presence of DTT (1.0 mM). Meanwhile, SOD activity was significantly raised by the addition of N-ethylmaleimide (NEM; 0.5 or 1.0 mM), a modifying reagent for the sulfhydryl reagent. Regucalcin (0.25 microM) caused a significant increase in SOD activity in the presence of NEM (1.0 mM). The effect of regucalcin in increasing SOD activity may not involve the sulfhydryl group of SOD. This study demonstrates that regucalcin has an activatory effect on SOD in the liver cytosol of rats. PMID- 15340236 TI - Comparison of the compositions of phospholipid-associated fatty acids in wild type and extracellular matrix tenascin-X-deficient mice. AB - Tenascin-X (TNX) is a member of the tenascin family of glycoproteins of the extracellular matrix. We previously showed that TNX regulates the synthesis of triglyceride and the composition of triglyceride-associated fatty acids. The aim of the present study was to determine whether TNX controls the synthesis of phospholipids and the composition of phospholipid-associated fatty acids by using TNX-deficient (TNX-/-) mice and TNX-overexpressing fibroblast cell lines. Thin layer chromatography of total lipids of the skin and sciatic nerves from wild type and TNX-/- mice revealed that the amounts of major phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), in wild-type and TNX /- mice are not different. Gas chromatography-mass spectrometry showed that the major fatty acid compositions of PC and PE in wild-type and TNX-/- mice are almost the same. Fibroblast cells stably overexpressing TNX also showed almost the same amounts of PC and PE and almost the same fatty acid compositions of PC and PE as those in mock-transfected cells. These results suggest that TNX regulates the amount of triglyceride and the composition of triglyceride associated fatty acids but not the amount and species of phospholipids or the composition of phospholipid-associated fatty acids. PMID- 15340237 TI - Preventive effect of tranilast on oleic acid-induced lung injury in guinea pigs. AB - Acute respiratory distress syndrome or acute lung injury (ARDS)/(ALI) involve the severe lung injury with pulmonary vascular hyper-permeability and hypoxemia induced by inflammatory reactions. Since ARDS/ALI carries high mortality, the development of new drugs against ARDS/ALI is required. We examined the effect of tranilast, an anti-allergic drug, on vascular hyper-permeability in the lungs and airways, and on hypoxemia, in oleic acid (OA)-induced acute lung injury, an animal model of ARDS/ALI. The increase in pulmonary and airway vascular permeability and the decrease in partial oxygen pressure of arterial blood induced by an intravenous injection of OA were drastically ameliorated by the oral administration of tranilast in a dose-dependent manner. This is the first report to prove that tranilast prevents pulmonary and airway vascular permeability and hypoxemia induced by OA. These results suggest that tranilast may be a candidate drug for the treatment of ARDS/ALI. PMID- 15340238 TI - Inhibitory effect of flavonoids on N-acetylation of 5-aminosalicylic acid in cultured rat hepatocytes. AB - 5-Aminosalicylic acid (5-ASA) is an effective drug for the treatment of ulcerative colitis and Crohn's disease. A large group of flavonoids was investigated for their inhibitory effects on the N-acetyl-conjugation of 5-ASA in rat hepatocytes and subcellular preparations. When added to cultured hepatocytes, some flavonoids inhibited the production of N-acetyl-5-aminosalicylic acid (5 AcASA) with potencies that depended on the specific structure of flavonoids. Among the flavonols, quercetin, kaempferol and galangin had inhibitory activity with a tendency to be more effective at increasing the number of hydroxyl substitutions in the B-ring. Flavones such as luteolin, apigenin and chrysin were as effective as the corresponding three flavonols above. 7,3',4'-OH flavone was more effective than other simple flavones such as 7-, 5-, 3-, 7,3-, 7,4'- and 3',4'-OH flavones. Isoflavones were relatively weak inhibitors. Taxifolin and catechins had little or no inhibitory effect. These data suggest that the presence of C7 hydroxyl substitution on the A-ring and the catechol group on the B-ring in the flavone structure is required for effective inhibitory activity. The inhibitory effect of flavonoids on N-acetyl-conjugation of 5-ASA was also examined by incubating 5-ASA with isolated liver cytosolic preparations. The active flavonoids in the cells inhibited the N-acetylation of 5-ASA in the cell free enzymatic preparations with a potency comparable to that for cultured rat hepatocytes. PMID- 15340239 TI - Grape seed extract prevents H(2)O(2)-induced chromosomal damage in human lymphoblastoid cells. AB - We examined the effects of grape seed extract (GSE) on chromosomal damage in two ways; induction on its own and prevention against treatment of reactive oxygen species (ROS). Chromosomal damage was evaluated by cytokinesis-block micronucleus assay (CBMN) in a human lymphoblastoid cell line, WIL2-NS cells. The GSE was composed of 89% proanthocyanidin with a degree of polymerization ranging from 2 to 15. GSE did not induce chromosomal damage in WIL2-NS cells at GSE concentrations up to 5 mg/l. In contrast, pretreatment with GSE dose-dependently prevented H(2)O(2)-induced chromosomal damage at an effective dose of 0.3 to 1 mg/l. A similar preventive effect of GSE was not detected in tert-butyl hydroperoxide-induced damage even at 5 mg/l. In a cell free system, GSE (<5 mg/l) directly scavenged H(2)O(2), but produced slight amounts of H(2)O(2) at higher concentrations (>50 mg/l). These results suggest that GSE is not genotoxic, but rather has an antigenotoxic effect against H(2)O(2) via direct scavenging action of H(2)O(2). PMID- 15340240 TI - Effects of trifluoromethyl ketones on the motility of Proteus vulgaris. AB - In the present study, we showed the inhibition of motility by trifluoromethyl ketone (TF) derivatives (1-8) in Proteus vulgaris (P. vulgaris) cultures. Among them, 1-(2-benzoxazoyl)-3,3,3-trifluoro-2-propanone (1) showed a much stronger inhibitory effect on the motility of P. vulgaris than other TF compounds at 10% MIC. Our results suggest the possibility of an inhibitory action of TF compounds on the proton motive forces by affecting the action of biological motor and proton efflux in the membranes, resulting in a reduction of the ratio of running and the increased number of tumbling and non-motile cells. PMID- 15340241 TI - Inhibitory effects of extracts from peels of Citrus natsudaidai encapsulated in hybrid liposomes on the growth of tumor cells in vitro. AB - Inhibitory effects of extracts from peels of Citrus natsudaidai (natsumikan) encapsulated in hybrid liposomes (HL) composed of L-alpha dimyristoylphosphatidylcholine and polyoxyethylene (20) sorbitan monolaurate on the growth of tumor cells were examined. The extracts with lower polar solvents inhibited the growth of B-16 mouse melanoma and human lung carcinoma cells, although the extracts with higher polar solvents showed no antitumor activity. In particular, the inhibitory effects of extracts with lower polar solvents encapsulated in HL were enhanced as compared with those of free extracts. Fluorescence microscopic analysis indicated that the HL including petroleum ether extracts induced apoptosis in B-16 mouse melanoma cells. On the other hand, the viability of normal human fibroblast cells was even less affected by the extracts of natsumikan. These results suggest that hydrophobic antitumor agents should be present in peels of natsumikan. PMID- 15340242 TI - Preventive effects of a traditional Chinese medicine (Sho-saiko-to) on endotoxin induced cytotoxicity and tumor necrosis factor-alpha production in J774A.1 Cells. AB - Sho-saiko-to, one of the most frequently prescribed Kampo medicines, is used clinically to treat chronic hepatitis and has shown confirmed clinical efficacy. The present study investigated whether Sho-saiko-to can suppress cytotoxicity and tumor necrosis factor (TNF)-alpha production in endotoxin-treated J774A.1 cells. Sho-saiko-to (10-20 microg/ml) did not affect the proliferation of J774A.1 cells, while a high concentration (50 microg/ml) of Sho-saiko-to induced a slight reduction in cell viability. Treatment with Sho-saiko-to (10-50 microg/ml) significantly inhibited endotoxin (10 microg/ml)-induced cytotoxicity in J774A.1 cells. In addition, Sho-saiko-to (20 microg/ml) suppressed TNF-alpha production by endotoxin (1 microg/ml)-activated J774A.1 cells. These findings suggest that the Kampo prescription Sho-saiko-to suppresses cytotoxicity or TNF-alpha production in macrophages treated with endotoxin and that it may be useful in improving septic shock symptoms. Sho-saiko-to may therefore protect against some of the various disturbances caused by endotoxins through its ability to inhibit TNF-alpha production in septic shock. PMID- 15340243 TI - HIV-1 inhibitory compounds from Calophyllum brasiliense leaves. AB - The hexane, acetone and methanol extracts of Calophyllum brasiliense leaves were fractionated following a three bioassay guide: high HIV-1 RT inhibition, low cytotoxicity on MT2 cells and high inhibition of HIV-1 IIIb/LAV replication. This led to the isolation of three anti HIV-1 dipyranocoumarins: calanolides A and B and soulattrolide. In contrast, other isolated compounds such as apetalic acid, isoapetalic acid, a structural isomer of isoapetalic acid, friedelin, canophyllol and amentoflavone were devoid of HIV-1 RT inhibitory activity. Calanolide C was also obtained as a natural product and showed moderate inhibitory properties. PMID- 15340244 TI - Effect of camellia oil on the permeation of flurbiprofen and diclofenac sodium through rat and pig skin. AB - The effect of camellia oil on the permeation of flurbiprofen (FP) and diclofenac sodium (DFS), used as model drugs, through rat and pig skin was examined. Two different types of camellia oil were used: one of them was purified by distillation and the other was purified by filtration without heating. The distilled camellia oil (DCO) and the filtered camellia oil (FCO) were applied to the skin as a pretreatment. Permeation of FP through the skins pretreated with FCO and DCO was enhanced, while that of DFS was suppressed. The effects of FCO were greater than those of DCO as far as enhancement and suppression were concerned. The effect of FCO on FP permeation could be due to oleic acid, one of the major components of FCO. On the other hand, FCO and oleic acid had opposite effects on the penetration of DFS. This result suggests that other active components which suppress the permeation of DFS may be present in FCO. Since the penetration-suppressing agents will be useful for skin care products, studies of such agents will be important in the future. PMID- 15340245 TI - Increased bioavailability of tacrolimus after rectal administration in rats. AB - The oral bioavailability of tacrolimus is low and varies considerably in humans due to first-pass metabolism by cytochrome P450 (CYP) 3A4 and the active efflux mediated by P-glycoprotein. This study was undertaken to elucidate the usefulness of rectal administration of tacrolimus as an alternative route to improve its bioavailability. Tacrolimus powder was suspended in a suppository base (witepsol H-15) and the tacrolimus suppository was inserted into the anus of the rats. For comparison, tacrolimus was suspended in 0.5% sodium methylcellulose solution and administered orally to rats. The dose of tacrolimus was fixed to 2 mg/kg. Blood samples were collected periodically up to 24 h after dosing, and tacrolimus concentrations were assayed by microparticle enzyme immunoassay. The whole blood concentrations of tacrolimus after rectal administration were much greater than those after oral administration. The C(max) and AUC(0-24 h) values after rectal administration were 3.9- and 6.9-fold greater than those after oral administration, respectively. These results clearly suggest a possibility that rectal administration of tacrolimus is capable of improving its bioavailability and cutting the costs of tacrolimus treatment. PMID- 15340246 TI - Pancreatic hamartoma. AB - Pancreatic hamartoma is a rare benign lesion and may be mistaken for a malignancy, as demonstrated by two cases. The first case was a 29-year-old man who presented with a 7-month history of intermittent upper abdominal pain, nausea and vomiting and a 15-kg weight loss. CT and MRI revealed a mass in the head of the pancreas. The second case was a 62-year-old man who presented with a 2-year history of intermittent abdominal pain, vomiting and a 25-kg weight loss. Although positron emission tomography was normal, CT revealed thickening of the duodenal wall and endoluminal ultrasonography revealed a tumour in the head of the pancreas. Both patients recovered from uneventful Kausch-Whipple pancreatoduodenectomy (in the first patient, it was pylorus-preserving), and in each case the histological diagnosis was hamartoma. Pancreatic hamartoma can present with vague, non-specific symptoms which, despite modern diagnostic tools, can be difficult to diagnose. Surgical resection with histopathological examination is required to confirm the diagnosis. PMID- 15340247 TI - Role of cAMP signaling in the mediation of dopamine-induced stimulation of GnRH secretion via D1 dopamine receptors in GT1-7 cells. AB - Pharmacologically increasing cyclic adenosine monophosphate (cAMP) levels in GT1 gonadotropin-releasing hormone (GnRH) cell lines increased the secretion of GnRH. Dopamine (DA) increased the GnRH secretion in GT1 cells via a DA receptor positively coupled to adenylate cyclase. We then asked whether inhibition of the DA-induced increase in cAMP would block the stimulatory effect of DA on GnRH release. Expression of the cAMP-specific phosphodiesterase (PDE4D1) was used in a genetic approach to inhibit the DA-induced increase in cAMP levels. Cells were infected with an adenovirus vector (Ad) expressing PDE4D1 (PDE-Ad) or, for controls, with an empty Ad (Null-Ad). Infection with the PDE-Ad completely blocked the forskolin-induced stimulation of GnRH secretion and [Ca2+]i and decreased the majority of the release of cAMP into the culture medium. In contrast, although PDE-Ad infection blocked virtually all of the DA-induced increase in extracellular cAMP, the release of GnRH and the increase in [Ca2+]i were only delayed for approximately 15 min. GT1 cells express the D1 DA receptor which is positively coupled to adenylate cyclase but not the D5 DA receptor. These data suggest that the initial phase of the DA-induced secretion of GnRH is dependent on an increase in cAMP levels. However, it appears that an additional non-cAMP-regulated signaling pathway is involved in the stimulation of GnRH release via the D1 DA receptor. PMID- 15340248 TI - Neurotransmitters involved in the opioid regulation of prolactin secretion at the end of pregnancy in rats. AB - Using a pharmacological approach, we explored potential mechanisms for the regulation of prolactin secretion by opioid peptides at the end of pregnancy in rats. On day 19 of pregnancy, intracereboventricular administration of the mu opioid receptor agonist (D-Ala2, NMe-Phe4, Gly-ol5)-enkephalin (DAMGO) or beta endorphin (beta-END) induced a dose-related increase in serum prolactin levels 30 min later. Pretreatment with the opioid antagonist naloxone abolished the increase induced by DAMGO injection. At lower doses, DAMGO and beta-END did not modify the 3,4-dihydroxyphenylacetic acid/dopamine ratio, but at higher doses, the mu-agonists evoked a significant increase of the dopaminergic activity as compared with saline control. The time course of the effects of beta-END (2.5 microg/rat) showed a higher increase in serum prolactin levels at 15 min than at 30 min after treatment. The 3,4-dihydroxyphenylacetic acid/dopamine ratio increased 15 min after beta-END administration and was even higher 30 min later. Neither the selective kappa-agonist U50,488H nor the selective delta-agonist (D Pen2, D-Pen5)- enkephalin were able to modify the serum prolactin levels at the doses studied. To evaluate potential neurotransmitters involved in the regulation of prolactin secretion at the end of pregnancy, we combined the administration of serotoninergic or GABAergic antagonists with the opioid agonist DAMGO. The serotonin 5-HT2 receptor antagonist ketanserin increased the serum prolactin levels and potentiated the effect of DAMGO. The intracerebroventricular administration of SR-95531 did not modify the serum prolactin concentration under basal conditions, but partially prevented the increase induced by DAMGO injection. The intracerebroventricular administration of the GABA(B) receptor antagonist phaclofen had no effect on the serum prolactin levels either in naive or DAMGO-treated rats. The present results support the proposal that activation of mu-opioid receptors stimulates prolactin secretion at the end of pregnancy. Although the exact mechanisms by which the opioid system modulates prolactin secretion at the end of pregnancy are unclear, these results suggest an interaction of the opioidergic system with serotoninergic and GABAergic systems, without ruling out a direct or indirect action on dopaminergic neurons. In conclusion, the opioid system may regulate prolactin secretion at the end of pregnancy through either stimulatory (present results) or inhibitory actions previously described. PMID- 15340249 TI - Functional expression of sodium-dependent vitamin C transporter 2 in human endothelial cells. AB - Since oxidative stress plays an important role in dysregulation of the microcirculation as well as the pathogenesis of atherosclerosis, therapeutic intervention with antioxidants has been speculated to prevent cardiovascular diseases. Ascorbic acid (AA) has been reported to improve endothelial function; however, its intracellular metabolic pathway has not been fully determined. Sodium-dependent vitamin C transporter (SVCT) types 1 and 2 were recently cloned. In the present study, we investigated whether SVCT-2 is functionally expressed in vascular endothelial cells and, if so, what factors modulate its activity. The uptake of AA into human umbilical vein endothelial cells (HUVECs) was examined by incubation with radiolabeled AA (14C-AA). AA was transported into HUVECs in a dose- and time-dependent manner. Replacement of sodium chloride with choline chloride in the medium suppressed the uptake of AA. RT-PCR revealed that HUVECs expressed SVCT-2 mRNA, but not SVCT-1. Transfection of HUVECs with the antisense oligonucleotide of SVCT-2 significantly suppressed the uptake of AA. Furthermore, tumor necrosis factor-alpha and interleukin-1beta inhibited the transport activity of AA. Thus, SVCT-2 is functionally expressed in human endothelial cells, and its activity is negatively regulated by inflammatory cytokines. Our findings might provide a new insight into understanding the treatment of cardiovascular diseases with AA. PMID- 15340251 TI - T regulatory cells in allergy and health: a question of allergen specificity and balance. AB - Anergy, tolerance and active suppression may not be independent events, but rather involve similar mechanisms and cell types in immune regulation. Induction of allergen-specific regulatory/suppressor T cells (T(Reg)) seems essential for the maintenance of a healthy immune response to allergens. Allergen-specific immunotherapy can induce specific T(Reg) cells that abolish allergen-induced proliferation of T helper 1 (Th1) and Th2 cells, as well as their cytokine production. T(Reg) cells utilize multiple suppressive mechanisms, interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta) as secreted cytokines and CTLA-4, PD-1, mTGF-beta, mIL-10, TGF-betaR and IL-10R as surface molecules. An important aspect of T(Reg) cells is the regulation of antibody isotypes and suppression of proinflammatory cells. IL-10 and TGF-beta secreted by T(Reg) cells skew production of IgE towards the noninflammatory isotypes, IgG4 and IgA, respectively. Furthermore, T(Reg) cells may directly or indirectly suppress effector cells of allergic inflammation such as basophils and eosinophils. In conclusion, induction of antigen-specific T(Reg) cells may redirect an inappropriate immune response against allergen or autoantigens with the help of a broad range of suppressor mechanisms. PMID- 15340255 TI - Single-step maternal serum screening for trisomy 21 in the era of combined or integrated screening. AB - Single-step maternal serum screening (MSS) in the first (1MSS) or second (2MSS) trimester at maternal age > or =35 years was evaluated in the North Belgian region Flanders, where difficulties are encountered in the general introduction of combined or integrated screening algorithms. The fetal aneuploidy screening database of General Medical Laboratory AML in Antwerp was searched for 2MSS tests between 1992 and 1999 (alpha-fetoprotein, beta-human chorionic gonadotropin (beta HCG) and unconjugated estriol, cut-off 1:300) and for 1MSS tests between 1999 and 2003 (free beta-HCG and pregnancy-associated plasma protein A, cut-off 1:85). At > or =35 years, the detection rate for trisomy 21 (DR) was 93.8% (15/16) for 2MSS and the screen-positive rate (SPR) was 24.5% (504/2061). For 1MSS, these figures were 85.7% (6/7) and 17.7% (109/615) respectively. To detect one trisomy 21, missed by MSS at > or =35 years of age, an additional number of 1,557 and 506 primary invasive procedures would be needed for 2MMS and 1MSS respectively. We conclude that the performance of both single-step 1MSS and 2MSS at maternal age > or =35 years in Flanders is excellent, even without the combination with ultrasound parameters or integration of first and second trimester parameters. The simplicity of both methods allows to consider them valuable options for fetal aneuploidy screening at advanced maternal age, until high quality combined or integrated screening is accessible to all pregnant women in Belgium. PMID- 15340256 TI - Effect of 12-month therapy with omega-3 polyunsaturated acids on glomerular filtration response to dopamine in IgA nephropathy. AB - BACKGROUND: Omega-3 polyunsaturated acids therapy is efficient in primary IgA nephropathy. It is unknown whether doses of omega-3 smaller than those given previously are still effective. The aim of the study was to examine the effect of omega-3 therapy on renal vascular function in relation to proteinuria and urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG). METHODS: 20 IgA patients aged 36.5 +/- 10.77 with creatinine clearance (Cr(cl)) 105.71 +/- 27.3 ml/min and proteinuria 3.31 +/- 2.01 g/24 h were given orally 810 mg EPA and 540 mg DHA daily for 12 months. Before and at the end of the study, 24-hour proteinuria, serum homocysteine, and Cr(cl) were measured. At the same time, renal vascular function was estimated as dopamine-induced glomerular filtration response (DIR). DIR was measured as: two 120-min lasting Cr(cl) (before and during 2 microg/kg b.w./min i.v. dopamine). RESULTS: The results obtained during follow-up were as follows (baseline vs. after therapy): DIR 14.9 +/- 16.4 vs. 30.3 +/- 14.3% (p < 0.01); urine protein 2.31 +/- 2.01 vs. 1.31 +/- 1.37 g/24 h (p < 0.01); (Cr(cl)) 105.71 +/- 27.3 vs. 103.9 +/- 20.9 ml/min (n.s.); NAG 8.3 +/- 1.8 vs. 6.0 +/- 1.2 U/g(creat) (p < 0.01), and homocysteine 16.2 +/- 3.15 vs. 13.8 +/- 2.6 micromol/l (p < 0.05). The only correlation found was linear correlation between basal DIR and DIR change (r = -0.570; p < 0.010) and basal NAG (r = -0.460; p < 0.50). CONCLUSIONS: Omega-3 supplementation is associated with the improvement of both renal vascular function and tubule function. PMID- 15340257 TI - Sporadic versus hereditary forms of colorectal cancer with the DNA microsatellite instability phenotype: to 'lump' or 'split'? PMID- 15340258 TI - Prognosis in DNA mismatch repair deficient colorectal cancer: are all MSI tumours equivalent? AB - Microsatellite instability (MSI) in colorectal tumours is the hallmark of defective DNA mismatch repair (MMR) and high level MSI can be detected in up to 15% of incident colorectal cancers. MSI in sporadic colorectal tumours is primarily due to epigenetic silencing of MLH1 while MSI is almost universal in tumours from HNPCC family members due to germline MMR gene mutation with loss or mutational inactivation of the second copy as a somatic event. There is evidence that tumour MSI is associated with a better outcome than the generality of large bowel malignancy. However, although MSI occurs in both sporadic colorectal cancer and in tumours arising in patients with germline MMR gene mutations, cancer survival should not be considered to be equivalent for these two groups with MSI tumours simply because both exhibit similarities in molecular phenotype. Here, we review the evidence on prognosis in patients with sporadic MSI tumours compared to those who have inherited a germline DNA MMR repair gene defect. In addition, we explore whether there are variables that afford opportunity to distinguish three groups on the basis of MSI status, namely: sporadic MSI tumours; MSI tumours in carriers of germline MMR gene defects; microsatellite stable (MSS) tumours. Differences in prognosis between these three groups is important because it underpins the rationale for surveillance and early identification of tumours in MMR gene carriers, as well as refining understanding of the influence of MSI on cancer progression. Furthermore, we discuss the effect of MSI on the effectiveness of chemotherapy regimens. PMID- 15340259 TI - HNPCC and sporadic MSI-H colorectal cancer: a review of the morphological similarities and differences. AB - Morphological features may serve as diagnostically useful markers of colorectal cancer (CRC) with the microsatellite instability-high (MSI-H) phenotype. The most important of these are lymphocytic infiltration, mucin secretion and poor differentiation. These features are apparent in both sporadic MSI-H CRC and CRC occurring in the context of hereditary non-polyposis colorectal cancer (HNPCC). There is now strong evidence that that the two principal subtypes of MSI-H CRC evolve through different pathways. Sporadic MSI-H CRC orginate within serrated polyps with BRAF mutation and DNA methylation while CRC in HNPCC arise within conventional adenomas in which there is frequent mutation of APC or beta -catenin and/or K- ras. These early differences in pathogenesis translate into multiple morphological distinctions in the cancers developing through the two pathways. Lymphocytic infiltration, tumour budding (de-differentiation), and co-existing adenomas are more evident in HNPCC while mucin secretion, poor differentiation, tumour heterogeneity and glandular serration, and co-existing serrated polyps are more evident in sporadic MSI-H CRC. Sporadic MSI-H CRC are also characterized by cytoplasmic eosinophilia and nuclei that are large, round, vesicular and contain a prominent nucleolus while in HNPCC the cytological features recapitulate the basophilia and nuclear characteristics of conventional adenomas. In practice, lymphocytic infiltration is the most sensitive marker of MSI-H status in both sporadic CRC and HNPCC. The crucial distinction between HNPCC and sporadic MSI-H CRC should be achieved by means of all available data including family history, age at onset of malignancy and molecular features. There is increasing evidence that genetic factors may predispose to DNA methylation. This can result in familial clustering of MSI-H CRC in which the underlying mechanism is methylation of hMLH1 rather than germline mutation. Morphological features can assist is distinguishing such families from bona fide HNPCC families which they closely mimic. PMID- 15340260 TI - Promoter hypermethylation frequency and BRAF mutations distinguish hereditary non polyposis colon cancer from sporadic MSI-H colon cancer. AB - BACKGROUND: Colorectal cancers resulting from defective DNA mismatch repair can occur in both hereditary non-polyposis colon cancer (HNPCC) and in the sporadic setting. They are characterised by a high level of microsatellite instability (MSI-H) and superficially resemble each other in that they are frequently located in the proximal colon and share features such as circumscribed tumour margins and tumour-infiltrating lymphocytes. However, significant differences can be demonstrated at the molecular level including widespread promoter hypermethylation and BRAF -activating mutations which occur significantly less often in HNPCC. AIMS: In this study, we sought to determine whether the presence of widespread promoter hypermethylation and BRAF mutations would exclude HNPCC. MATERIALS AND METHODS: We investigated the methylation status of four methylated in tumour markers (MINTs 1,2,12 and 31), and the promoter regions of 5 genes hMLH1, HPP1, MGMT, p16INK4A and p14ARF, in 21 sporadic MSI-H colorectal cancers and compared these with 18 cancers from HNPCC patients. The methylation status of CpG islands were determined by either methylation specific PCR (MSP) or combined bisulfite restricton analysis (COBRA). In addition we considered the BRAF mutation status of 18 HNPCC tumours and 19 sporadic MSI-H cancers which had been previously determined by RFLP analysis and confirmatory sequencing. RESULTS: Methylation of the promoter regions in target genes occurred less frequently within the HNPCC tumours (27% of analyses), compared with the sporadic MSI-H tumours (59% of analyses) (P < 0.001). Methylation of MINTs 1, 2, 12 and 31 occurred in 4% of analyses for HNPCC tumours contrasted with 73% for sporadic MSI H tumours (P < 0.001). BRAF mutations were detected in 74% of sporadic tumours but none of the HNPCC cancers tested. CONCLUSIONS: The total number of genes and MINTs methylated in HNPCC was lower than in MSI-H colorectal tumours. No HNPCC tumour showed evidence of widespread promoter hypermethylation or BRAF mutation suggesting this feature could be used as a discriminator between familial and sporadic cases. PMID- 15340261 TI - Evaluation of psychosocial effects of pre-symptomatic testing for breast/ovarian and colon cancer pre-disposing genes: a 12-month follow-up. AB - A prospective study of psychosocial consequences following predictive testing for inherited mutations in breast/ovarian and colon cancer susceptibility genes BRCA1, BRCA2, MLH1, and MSH2 was performed. Eighty-seven healthy women were tested for known family mutations and self-assessment scales were used to evaluate anxiety, depression and quality of life. Extensive pre- and post-test information was given. Questionnaires were responded before testing and four times after during the following year. A statistically significant decrease in anxiety mean scores over time was observed among the studied participants. The levels of depression in cancer genes carriers decreased over time while, surprisingly the levels in non-carriers increased. Compared to a normative Swedish sample all women tested showed similar levels of anxiety but women tested for breast cancer genes showed statistically lower levels of depression. Vitality dropped initially after disclosure of the testing of colon cancer genes carriers, followed by increasing levels. No change in vitality or in other quality of life parameters was seen in the other groups and the levels were similar to Swedish norm data. Most tested individuals were satisfied with the testing procedure including genetic counselling and testing and all of them but one would redo the testing. Healthy self-referred women going through predictive breast/ovarian or colon cancer gene testing, including extensive pre- and post-test information and support, in general, will not experience adverse psychological consequences. PMID- 15340262 TI - Correlations between phenotype and microsatellite instability in HNPCC: implications for genetic testing. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is widely considered to be a syndrome of defective mismatch repair (MMR). A major concern with genetic diagnosis of HNPCC is the variable, often low, percentage of pathogenic germline mutations that can be detected in MMR genes using common screening methods. The variable percentage of mutation detected is in part related to the sensitivity of conventional screening methods and may also depend on the heterogeneous genetics of HNPCC. Thus, identification of phenotypic criteria predictive of germline mutations in MMR genes may be helpful in efficient HNPCC genetic testing. Clinical diagnostic criteria, initially developed for HNPCC (e.g., Amsterdam I and II, or Bethesda criteria), can be used to clinically select patient candidates that carry germline mutations in MMR genes. More useful criteria were previously developed by analyzing families with germline MMR mutations. Using a complementary approach based on tumor microsatellite instability analysis, we confirm that the Amsterdam criteria are significantly better than the Bethesda criteria in predicting families with MSI-H tumors (P = 0.0227). Our results also suggest that a cutoff at < 50 years' mean age at diagnosis of HNPCC-related cancers (especially colorectal and endometrial cancer) may be an additional tool for the identification of families with defective MMR. Recent advances in MMR mutation screening are expected to improve detection of pathogenic MMR mutations in these families. Conversely, the high proportion of MSS tumors observed in our series of families with advanced age at cancer diagnosis is consistent with the low percentage of MMR mutations detected by previous studies in families with this phenotype. These families probably carry mutations in other genes that may or may not be related to MMR. Additional studies are necessary to clarify the molecular basis for HNPCC in families with MSS tumors. PMID- 15340263 TI - A homozygous MSH6 mutation in a child with cafe-au-lait spots, oligodendroglioma and rectal cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant condition due to heterozygous germline mutations in DNA mismatch repair genes, in particular MLH1, MSH2 and MSH6. Recently, a syndrome was recognized in which children develop haematological malignancies, solid tumours and signs of neurofibromatosis type 1 due to bi-allelic MMR gene mutations in MLH1, MSH2 and PMS2. Here we describe the child of healthy consanguineous parents who had cafe au-lait spots, oligodendroglioma, and rectal cancer. The patient was homozygous for the MSH6 mutation c.3386_3388delGTG in exon 5 which has a predicted pathogenic effect. Germline NF1 gene mutation testing was negative. The rectal tumour showed microsatellite instability and absence of MSH6 staining, whereas the brain tumour was MSI stable and showed normal immunohistochemical expression of MSH6. Apparently, not only MLH1, MSH2 and PMS2, but also MSH6 is involved in the syndrome of childhood cancer and signs of neurofibromatosis type 1. PMID- 15340265 TI - Impact of a cancer registry-based genealogy service to support clinical genetics services. AB - In collaboration with the network of genetics clinics in Scotland, a brief questionnaire was designed to gather data prospectively about the impact of information arising from pedigree research provided by Scottish Cancer Registry personnel. Pedigree research in Scotland is facilitated by access to public records of births, deaths, marriages, and historic census returns up to 1901, and enables the construction of accurate and extensive family pedigrees encompassing generations beyond the detailed knowledge of the proband. Subject to existing confidentiality guidelines, linkage of these pedigrees to cancer registration records results in a more comprehensive family history including the age at diagnosis of any cancer, multiple primary cancers, and cancers unreported from death certificates. Of 454 requests for pedigree research completed between 1 April 2002 and 31 March 2003, questionnaires were returned for 425 (94%). The information fed back to genetics clinics led to changes in family history, risk categorisation, and management in 41%, 30%, and 23% of cases, respectively. Management advice altered in both directions, that is, to institute active follow up and surveillance of clinic attendees and their relatives where none was previously envisaged, and viceversa. The interests of current and future generations of patients concerned about their familial risk of cancer will be served by measures which enable cancer registries to collect data that are as accurate and complete as possible. PMID- 15340264 TI - Genotyping possible polymorphic variants of human mismatch repair genes in healthy Korean individuals and sporadic colorectal cancer patients. AB - The genotypic consequences of numerous single-nucleotide variants in human mismatch repair genes are mostly undetermined. We examined 27 reported single nucleotide variants, rarely or ambiguously verified in a population-based study, to identify single-nucleotide polymorphisms (SNPs), haplotypes, and the genotype phenotype association in Korean populations of 330 healthy individuals, 107 sporadic colorectal cancer patients, and 107 of their first-degree relatives. Real-time PCR 5'-nuclease assays (TaqMan) MGB assay) were used to determine 24 single-nucleotide variants, and restriction fragment length polymorphism (RFLP) assays were used to determine 3 variants. Of these 27 variants, 4 (hMSH2 gIVS12 6, hMLH1 655, hMLH1 1151, and hMSH2 1168, in descending order) were identified as SNPs occurring in 4.5 to 53.1% of healthy individuals, with polymorphism levels of 0.023-0.3 (mean, 0.092). East Asian populations had an ethnic predilection for the hMLH1 1151 SNP. The genotype distribution for all four SNPs showed no association with sporadic colorectal cancer. Twenty-three variants were not identified in the Korean population, suggesting that fifteen of these variants are colorectal cancer-related mutations and eight are SNPs. Two haplotype patterns existed exclusively, but with rare frequency, in sporadic colorectal cancer patients. The hMLH1 655 allele was closely correlated with hMLH1 protein expression (P = 0.02), but none of the four SNPs was associated with clinicopathologic variables. Among the 27 single nucleotide variants of mismatch repair genes, 12 were suggestive of nonfunctional SNPs and 15 may be colorectal cancer-related mutations. Further verification in other ethnic groups may provide the genotypic and phenotypic significance of single nucleotide variants found in mismatch repair genes. PMID- 15340268 TI - [Investigation report on causes of radiation overexposure accident at Hirosaki national hospital and prevention of similar accident]. PMID- 15340269 TI - [Ethical principles and code of ethics]. PMID- 15340270 TI - [Ultrasonogram of pancreas and digestive tract]. PMID- 15340271 TI - [Challenge to Society Of Nuclear Medicine meetings]. PMID- 15340272 TI - [Know-how of the digital cine network construction]. PMID- 15340273 TI - [A study for the prevention of medical accidents for clinical examination in a Department of Radiology: part 3. risk cases due to CT, MRI, nuclear medicine and radiotherapy]. PMID- 15340274 TI - [A study for narrow beam dosimetry using a radiophotoluminescent glass rod dosimeter]. PMID- 15340275 TI - [A report of "Research for Radiography Techniques in X-ray Diagnosis"]. PMID- 15340276 TI - [Report on 19th short-time overseas study in the University of Chicago]. PMID- 15340277 TI - [X-ray protection construction standardization manual. Part 2]. PMID- 15340278 TI - [Optimization of the chest exposure condition with a contrast-detail phantom: evaluation of the flat-panel versus computed radiography systems]. AB - In this study, we evaluated the performance of a digital chest imaging system using a contrast-detail (C-D) phantom. In the initial step, 76 sample images of the C-D phantom were produced by changing the doses from 0.5, 0.75, 1.0, 1.25, 1.5, to 2.0 times the dose for a screen-film (S/F) system. The sample images were analyzed by five radiological technologists and two medical physicists, and the image quality figure (IQF) was determined. The quality of each image was examined, and appropriate doses were determined from the calculated IQF to obtain the same image quality for other digital chest imaging systems. The method of determining IQF from C-D phantom analysis was very useful for comparing image quality and determining radiographic techniques. PMID- 15340279 TI - [Evaluation of time resolution of the primary motor cortex by a single paradigm using a stimulation system]. AB - Many researchers are investigating brain function by using a block paradigm in fMRI measurement. However, time resolution has been limited because the block paradigm employed data collection by consecutive image acquisition after stimulation began. Because of this problem, the evaluation of early brain activity was not adequate. Therefore, a system that was able to take imaging from a high time dependence to stimulation in real time was necessary to evaluate early brain activity. It is necessary to obtain controlled image taking by an MRI device with an external trigger by creating a sequence design in the system controlling stimulation and a sequence designed to achieve the above. The execution of a single paradigm by the originally designed sequence thus becomes possible, and arbitrarily setting all processes from stimulation to image taking is possible. It is thought that the practical use of the single paradigm designed by the control system of stimulation enables real-time imaging for stimulation and provides time-dependent data. From this, evaluation of brain activity of the primary motor cortex is possible in the early stage by event-related data collection. In addition, "sank," a relative decrement in PO(2) was also an initial stage of BOLD, that is, an initial dip occurs as a transitory signal change by using a single paradigm that synchronizes with the control system of stimulation for the evaluation of early brain activity. Moreover, it is thought that taking images by a single paradigm that is synchronized with the control system of stimulation is indispensable for time and spatial elucidation of detailed brain activity, to evaluate hyper brain function. Therefore, the brain activity in the primary visual cortex at the early stage was clarified in this study by using a single paradigm designed to synchronize with the control system of stimulation by single-shot EPI. PMID- 15340280 TI - [Visual impression of photographic density of lung, thorax, and mediastinum in chest CT images: verification of optical illusion by visual subjective evaluation of medical images]. AB - Even if the brain in head CT images is shown physically at the same photographic density, optical illusion (assimilation, contrast, etc.) occurs and practical density can be observed psychologically differently. Therefore, in this study, in order to clarify the visual characteristics of lightness perception in chest CT images, I attempted to compare it with the visual impression of the photographic density of lung, thorax, and mediastinum, using visual subjective evaluation. The results were as follows. (1) In the case of the preferential window setting of lung, both thorax and mediastinum, which surround the lung, are recognized as a wide white frame with contrast, and the photographic density of the lung psychologically seems blackish as a result of the "picture frame effect". (2) In the case of the preferential window setting of thorax and mediastinum, the visual impression of the photographic density varies among observers. (3) In the case of the virtual double window setting of lung, thorax, and mediastinum, under the influence of both high-density and low-density areas in the radiological anatomy of thorax and mediastinum, the photographic density of lung psychologically appears whitish as a result of the "grouping theories" of lightness computation. Further, under the influence of original gray lung, the photographic densities of thorax and mediastinum psychologically appear to be whitish. PMID- 15340281 TI - [Development of automated measurement method for medial temporal lobe on CT images]. AB - Recent research has suggested that the measurement of regional atrophy in the structure of the medial temporal lobe is a promising way to discriminate Alzheimer-type dementia patients from healthy control subjects. The purpose of this study was to develop a technique to measure the medial temporal lobe automatically in axial CT images. Linear measurements of width of the inferior horns of the lateral ventricles, width of the medial temporal lobe, and the interuncal distance were performed. Area measurements of the inferior horns of the lateral ventricles were also performed. In the algorithm for the automatic measurement of the medial temporal lobe, brain contour, and sagittal plane were detected first, and the inferior horns of the lateral ventricles. Our method was applied to ten patients clearly without cerebral hemorrhage or infarct. The rates of accuracy of automated detection were 93% with the linear measurements and 75% with the area of the inferior horns. The rates were improved to 100% with the variable function of the threshold value. We suggest that this automated measurement method is both objective and simple enough to be used in routine clinical applications. PMID- 15340282 TI - [Measurement of gradation curve by the digital test pattern method in a computed radiography system]. AB - Computed radiography (CR) has many variations in gradation, and it is difficult to grasp all of them. We produced the Digital Test Pattern, a step chart made based on digital values that was created by personal computer using Binary Editor. Image data input in the CR system is replaced with the Digital Test Pattern data. A gradation curve is measuring for using the processed output image data in the CR system, such that we can determine the relation between input and output. We termed this the Digital Test Pattern method. Output characteristics after gradation processing are obtained by output signal value and input signal value. Input is known beforehand, and change in the gradation processing parameter is understood from the output pixel value. It is also possible to measure a gradation curve directly from photographic density. Measuring the relation of input and output enabled computer simulation of a gradation curve. These studies indicated the effectiveness of our proposed method in terms of accuracy and ease of use. PMID- 15340283 TI - [Evaluation of the number of sampling step angles in SPECT Images: comparison between FBP and OS-EM reconstruction algorithms]. AB - We compared filtered back projection (FBP) and ordered subset expectation maximization (OS-EM) reconstruction algorithms to examine the effects of sampling step angle on SPECT image reconstruction. Image sampling was compared between conditions of constant total sampling time (16 min) and constant sampling time (40 sec) for each step with angles of 3, 5, 7.5, and 10 degrees. Generally, OS-EM was superior to FBP in terms of image reconstruction. Changes in the sampling step angles did not significantly affect the quality of the reconstructed images with OS-EM insofar as acquisition counts were sufficient for evaluation. On the other hand, the quality of FBP images deteriorated with larger angles such as 7.5 and 10 degrees, and it was difficult to obtain stable results. In conclusion, OS EM improves the quality of images required for examination. PMID- 15340295 TI - [Effect of cariesogenic diet and age on selenium distribution between blood and mineralized tissues in rats]. AB - The distribution of Se in the blood and teeth (in the jaw and femoral bones) was evaluated using a new coefficient of relative radioactivity (RRA), representing the ratio of (75)Se in the teeth (bones) and blood. Changes in all RRA were studied during 1-192 hours after intraperitoneal injection of [(75)Se]selenate in one-month-old rats receiving common fodder; a biphasic pattern of changes was observed, with peaks 6 and 48 h and lowering 24 and 192 h postinjection. The first peak was mainly due to Se adsorption on hydroxyapatite, the second to chemo adsorption on it and Se binding to protein. In 3-month-old rats receiving common fodder the peak of all RRA was delayed to 12-48 h postinjection, the values decreasing by 192 h. The peak of specific activity (SA) of (75)Se protein in the bones was recorded 24 h and its decrease 192 h postinjection. In one-month-old rats receiving sucrose-casein diet during 2 months all RRA and SA after 6 and 12 h and part of RRA after 24 h were higher than in the control, that is, Se absorption by bones and teeth from the blood decreased. PMID- 15340296 TI - [Ultrastructure of gingival blood capillary endotheliocytes in patients with chronic catarrhal gingivitis in the presence of connective tissue dysplasia]. AB - Endotheliocyte structure in the gingival capillaries of patients with chronic catarrhal gingivitis (CCG) in the presence of connective tissue dysplasia and arterial hypertension differs from that in patients with CCG without somatic diseases, which essentially increase the permeability of capillary walls. PMID- 15340297 TI - [Mass spectral analysis of water in intact dental enamel in subjects with different caries resistance]. AB - Mass-spectrum analysis of specimens of intact dental enamel of subjects with different caries resistance was carried out. Products of gaseous phase released during constant heating of enamel were analyzed. Water mass spectrum was distinguished to be used in thermal programmed desorption analysis for the study of the organic constituent of dental enamel. PMID- 15340298 TI - [Location and intensity of dental caries in patients with osteoarticular tuberculosis]. AB - The intensity and location of carious lesions were studied in 140 patients with severe forms of osteo-articular tuberculosis aged 30-56 years. High intensity of carries and its frequent atypical location were observed during activation of the tuberculous process. Index of oral hygiene in patients with severe osteo articular tuberculosis was 4.55+/-1.6 vs. 2.53+/-0.10 in the control group; the intensity of dental caries was also higher. PMID- 15340299 TI - [Laboratory evaluation of the efficiency of dental root channel filling]. AB - Adhesive strength of gutta-percha stems connection to the root channel walls was evaluated. The pressure on dental root channel walls during tooth filling by the lateral condensation method was evaluated. The study was carried out on an Instron device by the pulling out method. The study showed that for stems Nos. 15 30 the retention strength in the root channel was about 2-fold higher than for stems Nos. 35-45. The results indicate that the value of stem pressure on root channel walls during tooth filling by the lateral condensation method can be considered critical with regard to the strength of the dental root proper. PMID- 15340300 TI - [Pressing problems in development of diagnostic methods for clinical periodontology]. AB - In this article are examining the requirements to diagnostic methods in periodontics. Author comprises clinical radiology and cytomorphometric methods in diagnostics of chronic gingivitis and initial periodontitis. PMID- 15340301 TI - [Optical coherence tomography in the evaluation of the oral cavity mucosa. Part II. Benign and malignant diseases]. AB - This paper is the last one in the series of publications on the investigation of the capability of Optical Coherence Tomography (OCT) in diagnostic of state of oral cavity mucosa. Part II demonstrates the diversity of optical images, different forms specific to leukoplakia, lichen planus and squamous cell carcinoma on the basis of the investigation of 56 pathologic lesions of 43 patients with precancer and cancer of oral cavity. The OCT sensitivity in malignancy detection in oral cavity is high - 83%, specificity is 98%, and accuracy is 81%. The kappa coefficient of interobserver agreement is 0.76. PMID- 15340302 TI - [Use of new solcoseryl-containing Diplan-denta C film in the treatment of injuries of the buccal mucosa]. AB - Clinical efficiency of bilayer adhesive dental film Diplan-denta C with solcoseryl in the treatment of postoperative wounds of the buccal mucosa and the effects of this film on the course of wound process were evaluated. The course of wound process in 39 patients with postoperative injuries of the buccal mucosa treated with Diplan-denta XD and Diplan-denta C films and traditional local therapy (0.05% chlorohexidine bigluconate solution and solcoseryl dental adhesive paste) was compared. In the study group Diplan-denta XD film with chlorohexidine was used for local therapy during the first 1-3 days after the injury and Diplan denta C film with solcoseryl was used in subsequent days until epithelialization; in controls irrigations of the oral cavity with chlorohexidine bigluconate solution (0.05%) were carried out during the first 1-3 days and applications of solcoseryl dental adhesive paste were made during subsequent days until epithelialization. The results indicate that the use of Diplan-denta C film optimized the treatment of the buccal mucosa wounds. PMID- 15340303 TI - [Correction of phagocyte functional activity in inflammatory focus in patients with deep phlegmons of the neck]. AB - High clotting activity (CA) of monocytes, low level of macrophage CA, and increase of proteolytic activity (PA) of neutrophils in the inflammatory focus predominate in patients with deep phlegmons of the neck at the peak of inflammatory process. Blood monocytes retain their anticlotting potential and macrophages possess CA during resolution of the inflammatory process. The neutrophil PA in the blood and lavage fluid was lower during the resolution phase in comparison with the peak of inflammation. Differentiated correction of phagocyte PA and LA in the inflammatory focus, predicted by laboratory findings, reduced the incidence of mediastinitis and improved the disease outcomes. PMID- 15340304 TI - [Leukinferon-induced changes in cytochemical parameters of the wound exudate neutrophilic granulocyte nucleus and cytoplasm in patients with maxillofacial phlegmons]. AB - Clinical and cytochemical study of the wound exudate neutrophilic granulocytes (NG) in patients with maxillofacial phlegmons treated traditionally and with leukinferon (immunomodulator) added to basic therapy demonstrated a positive clinical effect and pronounced stimulation of NG in the patients treated with leukinferon. The time course of cytochemical parameters of NG cytoplasm and chromatin activity was principally different in the two groups of patients. A new tissue index of NG was developed, giving an accurate integral picture of the total biological activity of the wound exudate NG. PMID- 15340305 TI - [Comparative evaluation of classical sinusotomy and endoscopic interventions on the maxillary sinus in odontogenic sinusitis with evaluation of the transporting function of ciliated epithelium and sinus aeration]. AB - In 2002 a total of 98 patients were operated on for odontogenic sinusitis, 49 of these were operated on by endoscopic methods and the rest by classical methods. The transporting function of the epithelium in the nasal cavity and maxillary sinus and the maxillary sinus aeration were evaluated. Pneumatization of the sinus was completely restored and the transporting function of the ciliated epithelium was less disturbed in the patients operated on by the endoscopic method, that is, the mechanisms of maxillary sinus cleansing from secretions were retained, which led to stable cure. PMID- 15340306 TI - [Clinical and morphological parallels in combined deformations of the nose]. AB - Analysis of clinical characteristics of 56 patients with combined deformations of the nose and morphological (histological and histochemical) changes in the nasal septal cartilage in 20 patients showed that common nasal deformations are complicated with age, impeding nasal respiration. The intensity of dystrophic changes in chondrocytes and intercellular cartilaginous matrix depends on the duration of posttraumatic or congenital deformation but not on its clinical severity. This necessitates earlier interventions, before the development of pronounced deformation of the whole nose and deterioration of nasal respiration. PMID- 15340307 TI - [Experience with utilization of the Michigan splint for temporary splinting during osteogingivoplasty]. AB - The experience of use Michigan splint for temporary splinting of the maxillary tooth at realization of flap surgery at 23 patients. Stages of clinical and laboratory procedures making the splint are described and analysed. Is appeared, that the reduction of mobility of teeth in the period following surgical intervention occurs on the average within 0.5-1 month faster, than with the patients after similar interventions, but without splinting. At the same time it is proved obvious, that the usage Michigan splint promotes normalization of functioning of muscles activity masticatory system and TMJ and helps in revealing of occlusal interferences. PMID- 15340308 TI - [Orthodontic treatment of patients after early dental implantation]. AB - Early dental implantation no later than 6 months after tooth removal under conditions of incomplete reparative osteogenesis was carried out in 62 patients (120 implantations). At the stage of orthodontic treatment a device for tooth restoration and fixation of artificial crown, suggested by the authors, was used. The efficiency of dentures supported by implants and natural teeth simultaneously was evaluated. The incidence of unfavorable outcomes of treatment was 5.8% after surgical stage of treatment and 11.7% after the patients were fitted with orthodontic constructions; the patients were observed during 5 years. The utilization of removable dentures supported by implants in patients with a complicated somatic status was analyzed. PMID- 15340309 TI - [Quality of life as the criterion of psychological status of patients with painful dysfunction of the temporomandibular joint]. AB - The quality of life in patients with painful temporomandibular dysfunction and the efficiency of serotonin antidepressants as an element of therapy of these patients were evaluated. The quality of life is decreased in this patient population; they develop hypothymic reactions. Use of serotonin-positive antidepressants appreciably improved the quality of life. PMID- 15340310 TI - [Transplantation of the liver right lobe from a related donor]. AB - From 1997 to 2003 transplantation of liver's right lobe from alive related donor was performed in 64 patients (28 men, 36 women) aged from 6 to 61 years (mean age was 22.6 +/- 3.2 years). Body weight of recipients ranged from 18 to 92 Kg (52.7 +/- 2.8 Kg on average). Indications for surgery were following: cirrhosis of liver due to Wilson's disease (34), primary sclerotic cholangitis (6), viral cirrhosis of liver (6), primary biliary cirrhosis (4), Bayler's disease (3), Caroly's disease (3), Budd-Chiary syndrome (3), secondary biliary cirrhosis (2), cirrhosis of liver due to deficient of alpha(1)-antitrypsin (1), hepatocellular carcinoma (1), total nodular transformation of liver (1). Intensive care before transplantation was needed for 68.75% potential recipients. Donors of liver's right lobe were relatives of patients similar to blood group. Age of donors ranged from 18 to 49 years, mean age was 37.9 +/- 1.4 years. Donors underwent right-sided hemihepatectomy, there were no complications. Lethal outcome in recipients was seen in 2 (3.1%) cases and was not associated with function of transplant. Lethal outcome in long-term period after transplantation was seen in 4 (6.5%) cases. All the others patients survived and followed from 1 to 73 months (23.8 +/- 2.4 months on average). Quality of life was good. Transplantation of liver's right lobe from related donor is the independent line in orthotopic transplantation of liver and ensure reliable results. PMID- 15340311 TI - [Autodermoplastic closure of defects of donor's zone after taking free revascularized autotransplants]. AB - Problems of autodermoplasty in donor's zone after taking of free revascularized autotransplants for closure of skin defect in 85 patients are regarded. All the transplants were complex. Free autodermoplasty was used in the zone of forearm -- 50 (58.8%) cases, foot - 22 (25.9%), posterior surface of thorax -- 9 (10.6%), and also in the zone of cnemis, lateral surface of femur, deltoid and scapular region -- one (1.2%) case each. In 52 (61.2%) cases free autoskin required for complete closure of defect in donor's zone and in 33 (38.8%) -- for partial closure. Region of taking of free autoskin was anterior-lateral surface of femur in majority of cases - 73 (85.9%). Split-thickness free autoskin was used the most often -- 79 (92.9%) cases. Graft retention was complete or partial in 77 (90.6%) cases, complete necrosis of free autoskin was seen in 8 (9.4%) clinical cases. It is concluded that method of free autodermoplasty is effective for closure of defects including in combination with microsurgical transplantation of tissues. PMID- 15340312 TI - [Comparative description of approaches in surgical treatment of aneurysms of descending thoracic aorta]. AB - Surgical treatment of patients with thoracic and thoracoabdominal aortic aneurysms is one of the most difficult and topical problems of up-to-date cardio vascular surgery. Right choice of surgical approach is important condition for effective surgery. This study is dedicated to comparative evaluation and definition of clear indication for choice of surgical approach in reconstruction of thoracic aneurysms. It is concluded that thoracotomy through 3(rd) intercost is preferable in isolated lesion of isthmus and proximal part of descending aorta. Approach through 5(th) intercost is not recommended in this localization of aneurysm. Prosthesis of all descending thoracic aorta (from arch to diaphragm) must be performed through double left-sided thoracotomy with single skin incision (left-sided double thoracotomy through 3(rd) and 6(th) intercosts from single S type skin incision). This approach always permits to perform surgical reconstruction of distal part of aortic arch, isthmus and all descending aorta, forms optimal conditions for creation of proximal and distal anastomosis. PMID- 15340313 TI - [Surgical treatment of patients with critical lower limb ischemia due to affection of arteries of the femoral-popliteal-tibial segment]. AB - Experience of surgical treatment of 325 patients with critical lower limb ischemia due to affection of infrainguinal vessels are analyzed. Comparative analysis of efficacy of direct, indirect and combined revascularizations was carried out, quality of life of patients in long-term period after surgery was evaluated. PMID- 15340314 TI - [Surgical treatment of major duodenal papilla neoplasm]. AB - Immediate and long-term results of pancreatoduodenal resections in 33 patients undergone surgery for adenocarcinoma of bile papilla from 1984 to 2003 were studied. Majority of patients had II and III stages of diseases. Correct diagnosis before surgery was made in 27 (81.8%) patients. Morphologic study revealed metastasis in 11 patients. Lethality was 9.1%, moreover last 18 pancreatoduodenal resections had no lethal outcomes. Long-term results were studied in 25 (83.3%) patients: 1-year survival was 90.4%, 3, 5 and 10 years survived 71.4, 61.2 and 51% patients respectively. Only metastatic lesion of lymph nodes and later stage of disease had negative influence on long-term results. Such factors as gender, diameter of carcinoma, lymphadenectomy and pylorus-saving variant of surgery did not influence on long-term results. It is concluded that pancreatoduodenal resection must be surgery of choice in the treatment of resected cancer of bile papilla. PMID- 15340315 TI - [Evaluation of results of surgical treatment of obliterating thrombangiitis in patients living in the North Arctic]. AB - Immediate and long-term results of surgical treatment of patients with obliterating thrombangiitis in patients living in Arctic North are analyzed. It is revealed that course of disease depends on length of patient's stay in Arctic North. These patients were younger, demonstrated more intensive progression of disease and more distal type of arterial lesion. Surgical treatment permitted to save 75.9% extremities in patients living in Arctic North with thrombangiitis and critical ischemia during 5 years after surgery that is worse than in patients living in Moscow region -- control group (86.1%). In long-term period more stable positive result was achieved after indirect revascularisation (save of extremity 78.3%). PMID- 15340316 TI - [Repeated surgeries on the thyroid gland in nodular euthyroid goiter]. AB - Medical histories of 214 patients hospitalized with diagnosis "recurrent nodular goiter" were studied retrospectively. Complex clinical, laboratory and instrumental examination was carried out in all the patients. Comparative analysis demonstrated that in the structure of recurrent goiter nodular colloid form dominated (84.1%), only in 42.1% cases repeated surgeries were indicated. Other patients (15.9%) were the ones underwent surgery for other thyroid diseases (hypertrophied form of autoimmune thyroiditis, follicular adenoma, thyroid cysts, thyroid cancer). Inadequate surgery and prophylactic therapy with iodine drugs were the main causes of recurrent goiter. Adequate iodine prophylaxis (potassium iodide 100-200 mg per day), complex examination of patients, correct indications for primary surgery and adequate surgical volume, valuable postoperative therapy (iodine drug, LT-4 if it is necessary) permit to reduce the risk of recurrent nodular (multiple-nodular, diffuse-nodular) goiter. Ways of researches for improvement of diagnosis, prophylaxis and treatment results of "recurrence nodular goiter" are projected. PMID- 15340317 TI - [Gastroduodenal bleedings in critical illness]. AB - Acute erosive lesions of upper parts of gastrointestinal tract with bleeding aggravate severe burn trauma, postoperative period after extensive surgeries and is the often component of polyorganic insufficiency syndrome. Gastric secretion in patients with severe burn trauma and neurotrauma was studied. Decrease of gastric secretory function due to central paresis of gastrointestinal tract and reflux of bile into stomach was seen in majority of patients with neurotrauma and acute gastric ulcers. On the contrary, in patients with burn shock increase of acid-secretory function of stomach was revealed. Schemes of prophylaxis and treatment of acute ulcers were developed. They included antacid therapy (for patients with increased secretion), regulators of motor-evacuatory function of stomach and intestine (for patients with paresis), drugs increased regenerative properties of mucosa, early enteral nutrition with balanced mixtures. This treatment in combination with hemostatic therapy and cure of main disease permitted to reduce number of gastroduodenal bleedings and lethality in these patients. PMID- 15340318 TI - [Diagnosis and treatment policy in trauma of duodenum]. AB - Experience of treatment of 70 patients aged from 17 to 71 years with trauma of duodenum (TD) was analyzed. Majority of them was men -- 57 (81.4%). Open TD were seen in 61.4% cases, closed -- in 38.6%. There were no pathognomonic symptoms. X ray examination of abdominal cavity is the important procedure permitted to suspect TD. Diagnostic algorithm in abdominal trauma for exclusion of TD before surgery was developed. Yellow-green imbibition, mass of air vesicles in retro abdominal space, hematomas in duodenal zone were the indications for revision of duodenum during surgery. Hematoma of duodenal wall was indication for it revision for exclusion of penetrating wound. In 55 (77.1%) patients with wounds penetrating into duodenum (or disruption of duodenum) suture of defects was performed. "Exclusion" of duodenum was performed in 20 (28.6%) patients when there was threat of sutures insufficiency. Lethal outcome was seen in 20 (28.6%) patients including 13 (18.6%) cases during first day due to combined injuries and massive blood loss. Developed algorithm and treatment policy permitted to improve results of this severe variant of abdominal trauma. PMID- 15340319 TI - [Lesions of retroperitoneal fatty tissue in destructive pancreatitis]. AB - Experience of examination and treatment of 102 patients with lesion of retroperitoneal space undergone surgery for pancreonecrosis is analyzed. Based of data of ultrasonic examination, computed tomography, surgery and autopsy laws of morphologic transformation and ways of spread of pathologic process into retroperitoneal space in pancreonecrosis were revealed. Developed scheme is important for substantiation of surgical policy. PMID- 15340320 TI - [Radical surgical treatment of early cancer of the stomach]. AB - Results of radical surgical treatment of 86 patients with early cancer of the stomach are analyzed. Among revealed macroscopic types of early cancer of the stomach (B0), type I (elevated) was seen in 26 (31%) patients, type IIa (superficial raised) - in 8 (9%), type IIb (superficial plane) - in 7 (8%), type IIc (superficial excavated) - in 20 (23%), type III (ulcerated) - in 25 (29%) patients. Adenocarcinoma was the prevalent tumor - 84 (98%) patients, in 1 (1%) case undifferentiated cancer was revealed and in 1 (1%) - carcinoid tumor. Tumor invasion was limited by mucosa in 49 (57%) patients, in submucosa - in 37 (43%). Metastases to lymph nodes were diagnosed in 4 (5%) patients. Subtotal proximal resection performed in 6 patients, subtotal distal resection - in 60, gastrectomy - in 20 patients. Lymphadenectomy corresponded to D1 in 30 cases, to D2 - in 51, to D3 - in 5 cases. Complications after surgery were seen in 21 (24%) patients. Cancer in the gastric stump was diagnosed in 3 (4%) patients 2, 4 and 6 years after primary subtotal distal resection of the stomach with D1 lymphadenectomy. All these patients underwent extirpation of gastric stump. Radical surgery for early cancer of the stomach must include extended lymphadenectomy D2 or more. PMID- 15340321 TI - [Surgical treatment of gastroesophageal reflux]. AB - One hundred and seventy-five patients with gastroesophageal reflux disease (GERD) were treated, 144 (82.2%) of them underwent surgery. In 106 (73.6%) patients laparotomy was used, in 38 (26.4%) -- laparoscopic surgeries. Transabdominal USE of the esophagus, stomach and duodenum, endoscopic ultrasonoraphy (EUS) of the esophagus were used for diagnosis of GERD in addition to standard tests. Based on EUS data, classification of severity of reflux-esophagitis was proposed. New method of laparoscopic creation of artificial lower esophageal sphincter (LES) with super thin (95, 45 mm) and super elastic filaments made of nikelid-titan alloy was developed in experiment and introduced into clinical practice. Thirty four laparoscopic surgeries were performed, 26 (76.4%) of them -- with creation of artificial LES. In this group 94.4% of patients demonstrated excellent and good results in long-term period. Pressure in the area of LES in these patients increased by 70%, on the average, after surgery. PMID- 15340322 TI - [Proteins of blood plasma in patients with pancreonecrosis]. AB - Dynamics of effective and general concentration of albumins and middle-size molecules in blood plasma of patients with severe pancreonecrosis is described. Significant differences of these data in operated and non-operated patients are revealed. A negative correlation between severity of pancreatic necrosis and concentrations of albumins, and a direct correlation between necrotic lesion and concentration of middle-size molecules in blood plasma were demonstrated. The above parameters are important for assessment of treatment efficacy, prognosis of postoperative complications and determination of indications to sanated relaparotomies in patients with pancreonecrosis. PMID- 15340323 TI - [Correction of bile outflow in complicated calculous cholecystitis]. AB - The method of low-traumatic atypical endoscopic papillosphincterotomy (EPST) with an antegradely inserted probe which permits to correct simultaneously bile outflow in choledocholithiasis and strictures of the terminal part of the common bile duct was used in 26 patients with cholelithiasis. Advantage of this surgical policy in complicated forms of calculous cholecystitis is possibility of one stage treatment. Comparative analysis of one-stage and routine two-stage treatment policies was carried out in 52 patients. EPST permits to avoid stage of retrograde pancreatocholangiography and papillosphincterotomy. Absence of technical difficulties due to EPST and a protective role of the probe which minimizes risk of acute pancreatitis are demonstrated. PMID- 15340324 TI - [Laparoscopy in diagnosis of abdominal injuries in patients with combined trauma]. AB - Importance of laparoscopy (LS) in diagnosis of intraabdominal catastrophe in patients with combined trauma (CT) is determined. Based on analysis of treatment results in 1,118 patients with CT it was revealed that real LS requirement does not exceed 10%, and it may influence negatively trauma outcome. Low diagnostic value of this method was revealed: sensitivity 98.4%, specificity 51%, accuracy 69.2%. It is concluded that LS is not a method of choice in diagnosis of intraabdominal injuries in patients with CT. PMID- 15340325 TI - [Surgical policy in complicated traumatic diaphragmatic hernias]. AB - Rare variants of surgical policy in different terms after traumatic diaphragmatic hernias complicated with necrosis and perforation of gastric walls and abdominal part of esophagus, ulcerous gastric bleedings, empyema of pleura, enzymatic gangrenous destruction of lung, cachexia are analyzed. Necessity of revision of diaphragm during surgery for diagnosis of it injuries is noted. In old diaphragmatic hernias and significant disposition of intraabdominal organs into pleural cavity thoracotomy or thoracophrenolaparotomy are recommended, in acute trauma when symptoms of abdominal injuries dominate laparotomy is expedient. In severe conditions minimal and organ-saving surgeries are recommended. Creation of "small stomach" from remains of its wall may be considered as alternative to traumatic gastrectomy. PMID- 15340326 TI - [Repeated surgeries in primary hyperparathyroidism]. AB - Surgery was performed in 161 patients with primary hyperparathyroidism (PHPT). Eight of 161 patients underwent repeated surgeries. In 6 patients persistent PHPT was associated with hyperplasia of parathyroid glands (PTG), in 1 -- with adenoma of PTG missed during the first surgery. In 4 of them hyperplastic PTG are located in anterior mediastinum, in 1 -- in posterior. In 2 patients with verified hyperplasia of PTG repeated surgeries were limited in the region of the neck. During repeated surgery in one patient recurrence of PTG cancer was diagnosed. Specific group consisted of 8 patients with non-severe disturbances of renal function which remained in early and long-term period after surgery. Levels of parathyroid hormone and ionized calcium were normal or insignificantly elevated. Dynamic observation was recommended for all patients of this group. PMID- 15340327 TI - [Insulin therapy in patients with myocardial ischemia and type II diabetes before and after myocardial revascularization]. AB - Fifty-two patients with coronary heart disease and type II diabetes have undergone myocardial revascularization. Correction of glycemia with insulin was carried out before, during and after surgery by standard schemes. Lethality and early postoperative purulent complications were compared. There were no significant differences in diabetic and non-diabetic patients. Good compensation of diabetes mellitus to recommend aorto-coronary bypass surgery in patients with type II diabetes on the same indications as for non-diabetic patients. PMID- 15340328 TI - [Quantitative myocardial perfusion assessment with magnetic resonance imaging in patients with coronary artery disease]. AB - AIM: To elucidate feasibility of the absolute myocardial blood flow (MBF), total coronary resistance (TCR), and myocardial blood flow reserve (MBFR) quantification using MRI in patients with coronary artery disease (CAD). MATERIAL AND METHODS: A total of 19 patients with angiographically documented CAD and 9 healthy subjects were studied by MRI using double-slice saturation-recovery Turbo FLASH sequence for monitoring myocardial first pass kinetics of Gd-DTPA-BMA at rest and during hyperemia (dipyridamole 0.56 mg/kg). The signal intensity curves were acquired within ROI for perfusion beds of the three main coronary arteries (LAD, LCX and RCA) and left ventricle cavity. Eighty five myocardial segments were included in final analysis (group 1 - supplied by <> coronary arteries, n=26; group 2 - supplied by arteries with non-significant diameter stenoses <50%, n=27; group 3 - supplied by arteries with significant stenoses >/=50%, n=32). Sixteen segments were revascularized subsequently (PCI or CABG). One-compartment model and slope-method were used for flow calculation. Myocardial and blood signal intensities were converted to concentration of Gd-DTPA-BMA according to the <> calibration curve. RESULTS: MBF was similar in groups at baseline (group 1 - 0.98+/-0.54, group 2 - 1.24+/-0.53 and group 3 - 1.28+/-0.48 ml/g/min) but significantly lower in group 3 during hyperemia (2.57+/ 1.23, 2.99+/-1.14 vs. 1,79+/-0.94 ml/g/min, p<0.05). MBFR (the ratio of flow during hyperemia to flow at baseline) was significantly lower in group 3 than in groups 2 and 1 (1.4+/-0.7 vs. 2.7+/-1.3 vs. 2.9+/-1.2, respectively, p<0.01). Receiver-operator characteristic analysis of MBFR (value /=50%). TCR (mean arterial pressure divided by flow) significantly decreased (78.8+/-42.2 vs. 41.3+/-17.3 mm Hg ? min ? g/ml, p<0.01), MBF accordingly increased (1.61+/-0.77 vs. 2.58+/-0.91 ml/g/min, p<0.01) during hyperemia and MBFR <> (1.3+/ 0.6 vs. 3.0+/-1.3, p<0.00l) in myocardial segments after revascularization. CONCLUSION: Absolute MBF and MBFR calculation by first-pass contrast perfusion MRI are feasible in patients with CAD before and after revascularization. PMID- 15340329 TI - [Sigmental right ventricular contractile function in patients with ischemic heart disease]. AB - Segmental right ventricular function was assessed by dobutamine stress echocardiography in 101 patients with ischemic heart disease and multiple coronary artery lesions. At rest local wall motion abnormalities were found in 69% of patients. Overall 505 segments of the right ventricle were analyzed (5 per patient). At baseline 34% of segments were asynergic (31% - hypokinetic and 3% akinetic), right ventricular wall motion score index was 1.38+/-0.04. Low dose dobutamine infusion resulted in decrease of portion of asynergic segments (to 6%) and lowering of wall motion score index (to 1.09+/-0.02, p<0.001 vs baseline). The use of stress doses of dobutamine was associated with appearance of ischemic changes of the right ventricle accompanied with typical anginal attacks and ST segment depressions; increases of portions of asynergic segments (up to 53% including 43% hypokinetic and 10% akinetic), and of patients with abnormalities of local contractility (up to 90%); rise of wall motion score index (up to 1.64+/ 0.05, p<0.001 vs low dose dobutamine). Segmental right ventricular wall motion abnormalities reflected mostly reversible myocardial dysfunction (hibernating myocardium was revealed in 28, scar - in 6, and zone at risk of ischemia - in 47% of all segments). Right ventricular myocardial dysfunction developed in patients with predominant involvement of the right coronary artery or anterior interventricular branch. PMID- 15340330 TI - [Anginal States provoked by psychogenic stimuli. To the problem of psychosomatic reactions in patients with ischemic heart disease]. AB - Two types of vulnerability of coronary vessels damaged by atherosclerosis were proposed basing on results of a study of 94 patients with psychosomatic reactions accompanied with ischemia or myocardial infarction. First variant (35 patients) was characterized by stable course of the disease (high effort angina without coronary catastrophes, episodes of unstable angina, etc.) and selective sensitivity to definite key affect of high personal value. Second variant (49 patients) was distinguished by pronounced clinical manifestations (effort angina with progressive lowering of exercise tolerance). Cardiovascular system displays universal susceptibility to negative emotions of various intensity. PMID- 15340331 TI - [Relationship between heart rate changes during reflex tests and heart rate variability in patients with recent myocardial infarction]. AB - BACKGROUND: Low heart rate variability (HRV) reflecting predominance of sympathetic tone is an independent predictor of sudden cardiac death after myocardial infarction (MI). We have previously shown that decreased heart rate (HR) response during reflex tests (breathing 6 per minute and Valsalva maneuver at the end of the first week after MI is also associated with elevated risk of sudden death during subsequent 2 years. AIM: To elucidate relationship between HR response to breathing 6 per minute and Valsalva maneuver, and HRV as standard non invasive method of assessment of autonomic regulation of the heart. MATERIAL: Tests with breathing 6 per minute and Valsalva maneuver were performed on 4-11 (median 8) days of MI in 188 patients (68.1% males) aged 34-75 (median 62) years, 93.6% of whom received beta-blockers. Parameters studied were difference between maximal and minimal HR during 1(st) min of controlled breathing (DHR) and Valsalva ratio (VR). Standard time- and frequency-domain HRV measures were calculated from 15-min ECG strips at bed rest just before tests and from 24-hour ECG recordings after tests. RESULTS: In patients with low values of DHR (<3.36) and VR (<1,13) most of HRV parameters were also significantly decreased. Most close association was found between low DHR and low SDNNi and LF power calculated from 24-hour Holter recordings, between low VR and low LF power at bed rest. CONCLUSION: Direct correlation between HR response to reflex tests and parameters of HRV allows to suggest that decreased HR response to controlled breathing and Valsalva maneuver in patients with recent MI also reflects presence of marked sympathetic predominance. PMID- 15340332 TI - [Efficacy of multifactorial preventive interventions in achievement of target levels of low density lipoprotein cholesterol in survivors of myocardial infarction]. AB - AIM: To assess Efficacy of multifactorial preventive Interventions in achievement of target levels of low density lipoprotein cholesterol in survivors of myocardial infarction. METHODS: Design of the study was open randomized comparative prospective. Two groups of patients with history of myocardial infarction were formed ("active intervention", n=88, "observation", n=48) and followed for 12 months. All interventions consisted in recommendatory and expository measures. Achievement of target low density lipoprotein cholesterol (LDLCH) levels, modifiable risk factors, blood lipids spectrum were assessed in 2 3, 5-6 and 12 months of observation. RESULTS: Overall effect of undertaken multifactorial intervention on modifiable risk factors was positive. By the end of the period of observation number of risk factors per patient decreased from 3.8+/-0.99 to 2.8+/-1.29 (p<0.05). However target levels parameters in many cases were not reached. About half of patients could not decrease consumption of saturated fats and cholesterol, nearly 25% continued to smoke. More than 55% maintained excessive weight and 64% had abdominal obesity. Among patients of "observation" group only 5 (4.2%) began to take lipid lowering drugs. In active intervention group average LDLCH lowering was 23%, 38% of patients received lipid lowering drugs and 44% (of those who completed the study) achieved target LDLCH levels. PMID- 15340333 TI - [Allele Frequency Analysis of Four Single Nucleotide Polymorphisms Locating in Promoter and 5'-Untranslated Regions of ABCAI Gene in Young Men - Survivors From Myocardial Infarction]. AB - Analysis of allele distribution of four single nucleotide polymorphisms (C-17G, C69T, G-191C and 319insG) of promoter and 5'-untranslated regions of the ABCA1 gene was carried out in a sample of 171 men, who had survived myocardial infarction before 45 years, and in controls. Two-fold increase of T69 and C-191 allele frequencies were observed in Russian population in comparison to Dutch one. While comparing allele and genotype distributions of the polymorphisms in the samples under study no statistically significant differences were found, so as no influence of different alleles on lipid spectrum data was observed. Role of polymorphisms under study appears to be insignificant in formation of genetic susceptibility to myocardial infarction in young men. PMID- 15340334 TI - [Effect of antiischemic therapy in immediate period after mammary coronary bypass surgery on left ventricular contractility and conduit blood flow. Randomized controlled study]. AB - AIM: To assess influence of 2-week course therapy with atenolol, nebivolol, and verapamil on systolic and diastolic left ventricular function and mammary coronary blood flow in early period after revascularization. MATERIAL: Patients (n=113) after mammary coronary bypass grafting. METHODS: Echocardiographic characteristics of systolic and diastolic left ventricular function, and blood flow through mammary coronary grafts were measured before and after treatment with study drugs. RESULTS: In placebo treated patients isovolumic relaxation time (IVRT) and late diastolic filling phase (A) increased, while velocity characteristics of mammary-coronary blood flow decreased. Administration of atenolol, nebivolol, and verapamil was associated with significant increases of left ventricular ejection fraction, stroke volume, cardiac output, IVRT, peak diastolic velocity, flow through mammary coronary grafts and their lumen diameter, and decrease of E/A ratio. CONCLUSION: Administration of atenolol, nebivolol, and verapamil in early postoperative period after mammary coronary bypass grafting improved systolic and diastolic function of the left ventricle, and blood flow through mammary coronary grafts. None of the studied drugs had significant advantage over others. PMID- 15340335 TI - [Dynamics of parameters of diastole and subendocardial right ventricular blood flow in adult patients after correction of atrial septal defect]. AB - The study of intracardiac hemodynamics and subendocardial blood flow in 55 adults with atrial septal defect confirmed practical value of assessment of myocardial contractility by long term catheterization of right cardiac chambers, construction of right ventricular function curves and "pressure-volume" diagrams. Simultaneous graphical registration with the help of computer diagnostic system represented character and direction of changes of controlled parameters during the period of observation as well as peculiarities of interventricular interaction after removal of isotonic right ventricular overload. Compared with traditional hemodynamic monitoring this approach to analysis of intracardiac hemodynamics provided real time assessment of cardiac function, facilitated elucidation of causes of cardiac output lowering and optimization of myocardial contractility regulation. PMID- 15340336 TI - [Antiremodeling Activity of Nebivolol in Patients With Essential Hypertension and Various Types of 4a/4b Polymorphisms of Endothelial NO Synthase Gene]. AB - Frequencies of alleles and genotypes of a polymorphic marker 4a/4b of eNOS gene and association of this marker with parameters of hemodynamics, signs of left ventricular hypertrophy and endothelial dysfunction were studied in 78 uzbek men aged 50.5+/-9.5 years with stage I-II essential hypertension. Allele 4b and genotype 4b/4b were found to be most frequent while genotypes 4a/4b and 4a/4a were associated with high degree of left ventricular hypertrophy, disturbances of 24-hour blood pressure profile and pronounced diastolic dysfunction. Three months of therapy with nebivolol exerted similar antihypertensive effect in all patients however only in patients with 4b/4b genotype it caused regression of left ventricular hypertrophy and normalization of endothelial vasoregulatory function. PMID- 15340337 TI - [Modern antidepressants in complex management of patients with hypertension and concomitant affective disorders]. AB - According to results of multiple studies depression and anxiety are found in more than 50% of patients with hypertension. Presence of affective disorders elevates risk of progression of hypertension. That is why complex therapy comprising antihypertensive drugs from various groups and antidepressants takes on higher and higher significance. Modern antidepressants produce no substantial cardiotoxic effects. Combination of an antihypertensive drug (angiotensin converting enzyme inhibitor captopril or beta-adrenoblocker metoprolol) with an antidepressant affects favorably clinical course of hypertension, 24-hour blood pressure profile, characteristics of intracardiac hemodynamics, structural and geometric left ventricular parameters, allows to achieve sufficient antihypertensive effect with acceptable tolerability. PMID- 15340338 TI - [Atorvastatin: lipid lowering activity, pleiotropic properties and efficacy in prevention of atherosclerosis and coronary heart disease]. AB - The review presents data on pharmacological properties of atorvastatin, its lipid lowering activity, and action on main mechanisms of atherogenesis. Favorable pleiotropic effects of atorvastatin such as normalization of endothelial function, antiinflammatory, antithrombotic and antioxidant activity contribute to stabilization of atheromatous plaques. Clinical trials revealing early and significant effects of the drug on main end points have demonstrated its high value for primary and secondary prevention of atherosclerosis. PMID- 15340339 TI - [The problem of optimization in cardiology]. AB - Optimization is understood as choice of the best version out of the competitive decisions. The strategy and tactics of prevention is presented from this point of view. Optimization of diagnostic investigations is viewed from the positions of continuum with quantitative assessment of results. Characteristics of loads and heart activity, as recommendations suggest, should be in agreement in stress tests. Optimization of dynamic investigations of biochemical markers is described on the model of acute coronary syndrome. Optimization of therapy is based on the determination of criteria of optimum. This statement is illustrated by the example of antihypertensive therapy. Optimization of pharmacotherapy must envisage the preservation of favorable effect of protective natural mechanisms in the form of autoregulation and adaptation to intermittent ischemia (preconditioning). PMID- 15340340 TI - [Long-Term Therapy With Indirect Anticoagulants in Patients With Nonvalvular Atrial Fibrillation (Prospective Follow-Up Study). Part II. Effectiveness and Safety]. AB - AIM: To assess efficacy and safety of long-term international normalized ratio (INR) guided therapy with acenocoumarol in patients with nonvalvular atrial fibrillation. MATERIAL: Patients (n=100) with nonvalvular atrial fibrillation and at least 1 risk factor of thromboembolic complications. METHODS: Ischemic strokes, episodes of systemic thromboembolism and hemorrhagic complications were registered during 3 years of treatment with acenocoumarol (target INR 2.0-3.0). RESULTS: Annual rates of ischemic strokes, hemorrhagic complications and major bleeding were 0.7, 14.4 and 1.1%, respectively. No episodes of thromboembolism were registered in patients with history of thromboembolic complications. Basing on data collected predictors of bleeding complications in patients with atrial fibrillation during long term treatment with acenocoumarol were elucidated. PMID- 15340341 TI - [Autoantibodies Against beta(1)-Adrenoreceptors in Patients With Cardiac Rhythm Disorders. Prevalence and Possible Role in Development of Arrhythmia]. AB - AIM: To assess prevalence of autoantibodies against beta(1)-adrenoreceptors (beta(1)-AR) in patients with arrhythmias of various etiology. MATERIAL: Patients with arrhythmias (n=110, including 59 patients with primary [idiopathic] electrical abnormalities, 33 - with chronic myocarditis and dilated cardiomyopathy [DCM]; 18 - with ischemic heart disease [IHD]) and healthy control subjects (n=20). METHODS: Antibodies against beta(1)-AR were measured in blood serum by direct immunoassay. Synthetic fragment containing 26 amino acids of beta(1)-AR second loop was used as antigen. RESULTS: Patients with primary electrical abnormalities and chronic myocarditis/DCM had similar prevalence of beta(1)-AR (49.1% and 54.5%, respectively), what was significantly higher than in controls (10%) and in patients with IHD (16.6%). These results provided evidence for the possible presence of an autoimmune process in the genesis of idiopathic arrhythmias. Among patients with idiopathic arrhythmias beta(1)-AR were found in 40% (10 of 25) of patients with ventricular tachycardia (VT), 63.6% (14 of 22) of patients with ventricular extrasystoles (VE), 41.6% (5 of 12) of patients with atrial fibrillation (AF). Among patients with chronic myocarditis and DCM beta(1) AR were found in 72.2% (13 of 18) of patients with VT, 28.5% (2 of 7) of patients with VE, 37.5% (3 of 8) of patients with AF. Among patients with idiopathic arrhythmias female sex and frequent respiratory viral diseases were more common in beta(1)-AR-positive compared with beta(1)-AR-negative patients. VT and left ventricular ejection fraction <40% were more common in beta(1)-AR-positive than beta(1)-AR-negative patients among those with chronic myocarditis and DCM. PMID- 15340342 TI - [Prevalence of electrocardiographical signs of right ventricular arrhythmogenic dysplasia]. AB - Right ventricular arrhythmogenic dysplasia (RVAD) is a state with high risk of sudden death in young patients. Early diagnosis of RVAD can facilitate sudden death prevention. AIM: To assess prevalence of electrocardiographical signs of RVAD among children with "idiopathic" tachyarrhythmias. MATERIAL: Patients without organic heart disease or overt noncardiac causes of arrhythmia aged 4-17 years (n=134, mean age 12+/-4.5 years, 56 girls and 76 boys) including 82 patients with >5000 extrasystoles VE 24 hours and 52 patients with ventricular tachycardia (VT). All patients had QTc interval <440 ms. METHODS: Twelve lead ECGs from all patients were analyzed for determination of morphology of arrhythmia and presence of "major" (epsilon wave and QRS duration >110 ms in V(1) V(3)) and "minor" (VT with left bundle brunch block - LBBB, VE >1000/24 hours, T wave inversion in V(2) and V(3)) diagnostic criteria for RVAD (W.McKenna, 1994; D.Corrado, 2000). RESULTS: ECG signs of RVAD were found in 28 of 58 (48.3%) of patients with VE and in 1 of 24 patients (4.2%) with atrial extrasystoles. Among patients with VE 27 (96.4%) had extrasystoles with LBBB morphology and 1 (3.6%) - with right bundle brunch block (RBBB) morphology. Combination of major and minor criteria sufficient for diagnosis of RVAD was found in 19% (8 of 42) of patients with VE with LBBB. Among 52 patients with VT 21 had polymorphic VT and 31 - monomorphic VT (16 with LBBB and 15 with RBBB). Epsilon wave was present in 56.3% (9/15) of patients with monomorphic VT and LBBB, in 4.8% (1/21) of patients with polymorphic VT and in none of the patients with monomorphic VT and RBBB. QRS duration exceeded 110 ms in 2 (12.5%), 2 (9.5%) and 0 patients among those with monomorphic VT and LBBB, polymorphic VT, and monomorphic VT and RBBB, respectively. Among patients with monomorphic VT and LBBB 37.5% (6/16) had combination of ECG criteria sufficient for diagnosis of RVAD. In 3 patients epsilon wave was registered in lead V(1) immediately prior to VT and disappeared after VT cessation. Implications. It can be suggested that presence of polymorphic VT or VE with LBBB morphology and specific ECG changes (first of all epsilon wave and QRS widening) distinguishes a group of patients with high risk of RVAD which requires thorough cardiological examination and dynamic observation. PMID- 15340343 TI - [Assessment of myocardial viability by dobutamine stress echocardiography in patients with ischemic heart disease]. AB - Detection of viable (hibernating) myocardium is necessary for determination of prognosis and tactics of treatment of patients with ischemic heart disease. For detection of viable myocardium and investigation of possibilities of its restoration 60 patients with ischemic heart disease (54 men, mean age 52+/-8 years) were examined before coronary artery bypass grafting or coronary angioplasty. Presence of viable myocardium was characteristic for patients with multivessel coronary artery disease (83%) with stenoses >90%, with well developed collateral circulation (81%). Sustained restoration of contractility of hibernating segments for 1 year after revascularization was noted in 70% of cases. Dobutamine stress echocardiography was found to have high diagnostic potential for detection of viable myocardium. PMID- 15340344 TI - [Effectiveness of early psychological rehabilitation among hospitalized patients with ischemic heart disease]. AB - The subjects of the study were men and women (n=418) suffering from myocardial infarction or angina pectoris and hospitalized in the Cardiological Clinic of Kaunas Medical University during May-June, 2001. In order to assess the possibility to apply measures of psychological rehabilitation in a hospital setting, patients were invited to participate in three group sessions of relaxation (achieved by visualizing certain images). Effectiveness of psychological intervention was assessed by measuring statements indicating presence/absence of type A behavior pattern, depressiveness, and anxiety. Blood pressure was measured twice during each session. 38.3% of men and 35.15% of women came to the first session, while all three sessions were attended by 14.2 and 7.8% of men and women, respectively. Patients did not participate due to subjective (negative attitude towards psychological measures) and objective (hospital regimen: treatment and diagnostic procedures, etc) reasons. Average age of participants of three sessions was 56.9+/-10.31 for men, and 64.4+/-8.91 for women (p<0.02). After three group sessions such indices of type A behavior pattern, depressiveness, and anxiety as nervous tension, impatience, being in a hurry, fatigue, worrying, pessimism, anxiety became lower as compared with those at the first session. PMID- 15340345 TI - [Relationship Between Lipid Lowering and Changes of Hemostasis and Inflammation During Use of Statins in Non-ST Elevation Acute Coronary Syndrome]. AB - AIM: To analyze relationship between changes of lipid levels and parameters of hemostasis and inflammation in a previously reported comparative study of some biological effects of pravastatin and atorvastatin in patients with non ST elevation acute coronary syndrome (NSTEACS). METHODS: Ninety aspirin and heparin treated patients with NSTEACS were randomized to open pravastatin 40 mg/day (n=31) and atorvastatin 10 (n=30) or 40 mg/day (n=29). At baseline, on days 7 and 14 we measured levels of thrombin-antithrombin complex (TAT), prothrombin fragments 1+2 (F1+2), D-dimer, von Willebrand factor (vWF) and C-reactive protein (CRP) and assessed platelet aggregation. Spearman correlation coefficients were calculated for mean deltas of all parameters {[(baseline - day 7) + (baseline - day 14)]/2} for all patients. The patients were divided into quartiles according to absolute LDL CH lowering by day 14. RESULTS: Levels of LDL and total CH significantly decreased in all groups (atorvastatin 40 mg/day >> atorvastatin 10 mg/day > pravastatin). Contrary to pravastatin the use of atorvastatin was associated with increases of F 1+2, TAT, and decrease of vWF. Platelet aggregation decreased only in atorvastatin 40 mg/day group (p<0.05) and CRP decreased in combined atorvastatin group (p<0.05). Most pronounced relationship existed between changes of lipid levels and vWF. Lowering of total and LDL CH correlated positively with decreases of vWF (r=0.25, p=0.018, and r=0.23, p=0.032, respectively) and quartile analysis showed that vWF decreased only in patients with marked LDL CH lowering (quartiles 3, 4). Negative correlation was found between changes of total CH and those of TAT (r=-0.22, p=0.039), and between changes of CRP and HDL CH (r=-0.24, p=0.03). TAT and F 1+2 levels significantly increased in those patients in whom LDL CH level <2.5 mmol/l was achieved by day 14 and tended to decrease in other patients. No relationship was found between changes of lipids and platelet aggregation. CONCLUSION: Rapid (in 14 days) changes of some parameters of hemostasis occurring during treatment of patients with NSTEACS with various statins were related to degree of lipid lowering possibly irrespective of a statin used. Most evident was relationship between changes of lipids and von Willebrand factor. PMID- 15340346 TI - [Impairment of cardiac sympathetic function in patients with acute myocardial infarction and unstable angina]. AB - AIM: To assess the state of sympathetic innervation of the heart in patients with acute coronary syndrome. MATERIAL: Patients with Q-myocardial infarction (MI, n=36), non-Q-MI (n=13), and unstable angina (UA, n=9). METHODS: Each subject underwent single-photon emission computed tomography (SPECT) and planar scintigraphy using iodine-123 metaiodobenzylguanidine ((123)I-MIBG) for assessment of cardiac sympathetic function. We analyzed early (15 minutes) and delayed (4 hours) images after (123)I-MIBG administration. Resting (99m)Tc-MIBI myocardial scintigraphy was performed for evaluation of myocardial perfusion. Location, extent (%) and severity (Un.) of defects were determined using <> program. (123)I-MIBG did not accumulate in myocardium of 3.5% patients. All other patients demonstrated (123)I-MIBG accumulation defects. These defects were colocolized with (99m)Tc-MIBI uptake abnormalities. However both extent and severity of sympathetic innervation defects exceeded those of perfusion defects in all patients. Mean extent and severity of sympathetic neuronal damage areas were the greatest in patients with Q-MI (41+/-8% and 1119+/-377 Un. respectively) and the least in patients with UA (22+/-12% and 602+/-353 Un., respectively). On the contrary the mean extent of areas with sympathetic endings dysfunction but normal perfusion was the largest in UA group and the least in Q-MI group (18+/-11 and 10+/-7%, respectively, p<0,05). CONCLUSION: Impairment of cardiac sympathetic function in patients with acute coronary syndrome could be detected by SPECT with (123)I-MIBG. Locations of (123)I-MIBG and (99m)Tc-MIBI defects were similar but sympathetic dysfunction areas were larger than areas with reduced perfusion. This result suggests higher sensitivity of sympathetic endings to ischemia compared with cardiomyocytes. Myocardial areas with sympathetic endings dysfunction but normal perfusion can be defined as myocardium at risk. PMID- 15340347 TI - [Postinfarction left ventricular remodeling in patients with hypertension]. AB - 1-3 years after myocardial infarction 140 patients (66 with - group 1, and 74 without - group 2 - hypertension before infarction) were examined by echocardiography. In group 1 concentric remodeling was found in 71.4% of patients with postinfarction scar occupying 50 years, but it remains unclear whether ILD precedes lung cancer or vice versa. In this review, we examine the epidemiology of ILD and the basis for its association with lung cancer. PMID- 15340373 TI - Treatment of non-small-cell lung cancer: a perspective on the recent advances and the experience with gefitinib. AB - Worldwide, non-small-cell lung cancer (NSCLC) is a leading cause of cancer related mortality and, until screening detects early disease, treatment for the majority of patients will consist of radiation therapy, chemotherapy or combinations thereof. Modern mono and doublet chemotherapy regimens have translated into modest increases in life expectancy and improved quality of life, but at the expense of systemic and pulmonary adverse events (AEs). There is a great unmet need to provide effective therapy for advanced NSCLC that does not have the toxicity burden of conventional chemotherapy and radiotherapy. Novel drugs that inhibit a range of growth factor receptors, such as the epidermal growth factor receptor tyrosine kinase inhibitors gefitinib ('Iressa') and erlotinib ('Tarceva') or the monoclonal antibody cetuximab ('Erbitux'), have recently been evaluated. Having demonstrated antitumour activity and rapid symptom improvement in pretreated patients with advanced NSCLC, gefitinib was approved in the USA, Japan and other countries. Gefitinib is well tolerated with a low incidence of grade 3/4 AEs. Interstitial lung disease has been reported in a small number of patients receiving gefitinib, although this may be attributed to other treatments and conditions. Nevertheless, although the use of novel treatments requires vigilance for unexpected AEs such as pulmonary toxicity, in this area of high unmet clinical need, the benefits outweigh the risks in patients for whom no other proven effective treatment exists. PMID- 15340374 TI - Interstitial lung disease associated with drug therapy. AB - Drug-associated interstitial lung disease (ILD) is not uncommon, with diverse patterns ranging from benign infiltrates to the potentially fatal acute respiratory distress syndrome. As acute respiratory failure due to drug associated ILD has an unpredictable onset and rapid time course, establishing a diagnosis is often difficult. An accurate diagnosis is based on clinical, radiological (including high-resolution computed tomography) and histological manifestations, although is often only possible by exclusion. Cancer chemotherapy is commonly associated with acute disease that, on pathology, is often diffuse alveolar damage. Furthermore, a combination of drugs with or without radiotherapy can increase the risk of ILD. This article reviews treatments for non-small-cell lung cancer (NSCLC) that are associated with the development of ILD and how systematic evaluation of the possible role of these drugs in ILD is warranted. A difference between Japan and the rest of the world in reporting rates of ILD when gefitinib ('Iressa') has been used in advanced NSCLC is also discussed. However, the difference remains unexplained, leaving important epidemiological and mechanistic questions. PMID- 15340375 TI - Diagnosis and management of drug-associated interstitial lung disease. AB - Symptoms of drug-associated interstitial lung disease (ILD) are nonspecific and can be difficult to distinguish from a number of illnesses that commonly occur in patients with non-small-cell lung cancer (NSCLC) on therapy. Identification of drug involvement and differentiation from other illnesses is problematic, although radiological manifestations and clinical tests enable many of the alternative causes of symptoms in advanced NSCLC to be excluded. In lung cancer patients, high-resolution computed tomography (HRCT) is more sensitive than a chest radiograph in evaluating the severity and progression of parenchymal lung disease. Indeed, the use of HRCT imaging has led to the recognition of many distinct patterns of lung involvement and, along with clinical signs and symptoms, helps to predict both outcome and response to treatment. This manuscript outlines the radiology of drug-associated ILD and its differential diagnosis in NSCLC. An algorithm that uses clinical tests to exclude alternative diagnoses is also described. PMID- 15340376 TI - Understanding the mechanisms of drug-associated interstitial lung disease. AB - Drugs have been implicated in lung injury as a result of direct pharmacological action, persistence or metabolism in the tissue, or via the production of a reactive metabolite or metabolites. The result of this apparent drug-associated injury ranges from cellular dysfunction through to cell death (apoptosis) and alteration of repair mechanisms that are essential in replacing critical tissue elements and function. There is limited knowledge on how timing of drug administration or drug interactions may interfere with the repair mechanisms or modulate the expression of pulmonary toxicity. Chemotherapeutic drugs and novel agents, such as those targeting the epidermal growth factor receptor (EGFR), appear to affect both normal and neoplastic cells. However, unlike chemotherapy, where the actions are systemic and directly as a result of biotransformation or cell injury, it has been postulated that effects of EGFR-targeting agents are more likely to be focused on epithelia via a pharmacological effect. Furthermore, risk factors for the development of adverse pulmonary reactions, as well as biological markers indicating incipient toxicity, need to be prospectively identified. Proteomics, through the identification of >/=1000 proteins or peptides in blood samples, will hopefully identify candidates for this role. PMID- 15340377 TI - From structure to disease: the evolving tale of aquaporin biology. AB - Our understanding of the movement of water through cell membranes has been greatly advanced by the discovery of a family of water-specific, membrane-channel proteins - the aquaporins. These proteins are present in organisms at all levels of life, and their unique permeability characteristics and distribution in numerous tissues indicate diverse roles in the regulation of water homeostasis. The recognition of aquaporins has stimulated a reconsideration of membrane water permeability by investigators across a wide range of disciplines. PMID- 15340378 TI - The ABC's (and XYZ's) of peptide sequencing. AB - Proteomics is an increasingly powerful and indispensable technology in molecular cell biology. It can be used to identify the components of small protein complexes and large organelles, to determine post-translational modifications and in sophisticated functional screens. The key - but little understood - technology in mass-spectrometry-based proteomics is peptide sequencing, which we describe and review here in an easily accessible format. PMID- 15340379 TI - Plasmodesmata as a supracellular control network in plants. AB - The evolution of intercellular communication had an important role in the increasing complexity of both multicellular and supracellular organisms. Plasmodesmata, the intercellular organelles of the plant kingdom, establish an effective pathway for local and long-distance signalling. In higher plants, this pathway involves the trafficking of proteins and various forms of RNA that function non-cell-autonomously to affect developmental programmes. PMID- 15340380 TI - Cell signalling and the control of pre-mRNA splicing. AB - The transcripts of most metazoan protein-coding genes are alternatively spliced, but the mechanisms that are involved in the control of splicing are not well understood. Recent evidence supports the potential of both extra- and intracellular signalling to the splicing machinery as a means of regulating gene expression, and indicates that this form of gene control is widespread and mechanistically complex. However, important questions about these pathways need to be answered before this method of post-transcriptional regulation can be fully appreciated. PMID- 15340381 TI - The SCF ubiquitin ligase: insights into a molecular machine. AB - Ubiquitin ligases are well suited to regulate molecular networks that operate on a post-translational timescale. The F-box family of proteins - which are the substrate-recognition components of the Skp1-Cul1-F-box-protein (SCF) ubiquitin ligase - are important players in many mammalian functions. Here we explore a unifying and structurally detailed view of SCF-mediated proteolytic control of cellular processes that has been revealed by recent studies. PMID- 15340382 TI - Apoptosis and genomic instability. AB - Genomic instability is intrinsically linked to significant alterations in apoptosis control. Chromosomal and microsatellite instability can cause the inactivation of pro-apoptotic pathways. In addition, the inhibition of apoptosis itself can be permissive for the survival and ongoing division of cells that have failed to repair DNA double-strand breaks, experience telomere dysfunction or are in an abnormal polyploid state. Furthermore, DNA-repair proteins can regulate apoptosis. So, genomic instability and apoptosis are intimately linked phenomena, with important implications for the pathophysiology of cancer. PMID- 15340383 TI - Metabolite profiling: from diagnostics to systems biology. AB - The concept of metabolite profiling has been around for several decades, but only recent technical innovations have allowed metabolite profiling to be carried out on a large scale - with respect to both the number of metabolites measured and the number of experiments carried out. As a result, the power of metabolite profiling as a technology platform for diagnostics, and the research areas of gene-function analysis and systems biology, is now beginning to be fully realized. PMID- 15340384 TI - Pharmacogenomics: bench to bedside. AB - Pharmacogenetics is the study of the role of inheritance in inter-individual variation in drug response. Since its origins in the mid-twentieth century, a major driving force in pharmacogenetics research has been the promise of individualized drug therapy to maximize drug efficacy and minimize drug toxicity. In recent years, the convergence of advances in pharmacogenetics with rapid developments in human genomics has resulted in the evolution of pharmacogenetics into pharmacogenomics, and led to increasing enthusiasm for the 'translation' of this evolving discipline into clinical practice. Here, we briefly summarize the development of pharmacogenetics and pharmacogenomics, and then discuss the key factors that have had an influence on - and will continue to affect - the translation of pharmacogenomics from the research bench to the bedside, highlighting the challenges that need to be addressed to achieve this goal. PMID- 15340385 TI - Technology platforms for pharmacogenomic diagnostic assays. AB - Rapid advances in the understanding of genomic variation affecting drug responses, and the development of multiplex assay technologies, are converging to form the basis for new in vitro diagnostic assays. These molecular diagnostic assays are expected to guide the therapeutic treatment of many diseases, by informing physicians about molecular subtypes of disease that require differential treatment, which drug has the greatest probability of effectively managing the disease, and which individual patients are at the highest risk of experiencing adverse reactions to a given drug therapy. This article reviews some of the relative strengths and limitations of the most widely used technologies and platforms for such assays. PMID- 15340386 TI - Translation of pharmacogenomics and pharmacogenetics: a regulatory perspective. AB - Pharmacogenomics and pharmacogenetics provide methodologies that can lead to DNA based tests to improve drug selection, identify optimal dosing, maximize drug efficacy or minimize the risk of toxicity. Rapid advances in basic research have identified many opportunities for the development of 'personalized' treatments for individuals and/or subsets of patients defined by genetic and/or genomic tests. However, the integration of these tests into routine clinical practice remains a major multidisciplinary challenge, and even for well-established biomarkers there has been little progress. Here, we consider this challenge from a regulatory perspective, highlighting recent initiatives from the FDA that aim to facilitate the integration of pharmacogenetics and pharmacogenomics into drug development and clinical practice. PMID- 15340387 TI - The endocannabinoid system and its therapeutic exploitation. AB - The term 'endocannabinoid' - originally coined in the mid-1990s after the discovery of membrane receptors for the psychoactive principle in Cannabis, Delta9-tetrahydrocannabinol and their endogenous ligands - now indicates a whole signalling system that comprises cannabinoid receptors, endogenous ligands and enzymes for ligand biosynthesis and inactivation. This system seems to be involved in an ever-increasing number of pathological conditions. With novel products already being aimed at the pharmaceutical market little more than a decade since the discovery of cannabinoid receptors, the endocannabinoid system seems to hold even more promise for the future development of therapeutic drugs. We explore the conditions under which the potential of targeting the endocannabinoid system might be realized in the years to come. PMID- 15340388 TI - Nanosuspensions in drug delivery. AB - A surprisingly large proportion of new drug candidates emerging from drug discovery programmes are water insoluble, and therefore poorly bioavailable, leading to abandoned development efforts. These so-called 'brickdust' candidates can now be rescued by formulating them into crystalline nanosuspensions. In the process of overcoming issues involving solubility, additional pharmacokinetic benefits of the drugs so formulated have come to be appreciated. As such, insolubility issues of the past have provoked a paradigm change, which now offers novel solutions for innovative drugs of the future. PMID- 15340389 TI - Paul Ehrlich: magister mundi. AB - Paul Ehrlich, a founding father in fields across a wide range of medical sciences, was born 150 years ago. To mark this anniversary, this article gives an overview of his life, highlighting his major scientific contributions and paying tribute to Paul Ehrlich the man. PMID- 15340390 TI - Biochemical mechanisms of drug action: what does it take for success? AB - Drug discovery is extremely difficult. There are many unanticipated scientific, medical and business challenges to every drug discovery programme. It is important to increase our understanding of the fundamental properties of effective drugs so that we can anticipate potential problems in developing new agents. This article addresses potential drug discovery and development risks associated with the biochemical mechanism of drug action, and proposes simple rules to minimize these risks. PMID- 15340391 TI - A systematic review of antidepressant placebo-controlled trials for geriatric depression: limitations of current data and directions for the future. AB - Depression in the elderly is a major public health problem as untreated depression adversely impacts comorbid illnesses. It is important to develop safe and effective antidepressant therapies for older individuals. We performed a systematic review of all published randomized, placebo-controlled antidepressant medication trials in populations over age 55 years. Papers were obtained via MEDLINE (1966-August 2003) and PSYCINFO (1872-August 2003). Unpublished trials, trials examining nonpharmacologic interventions, and papers reporting post hoc analyses were not included in this review unless they provided new insights. A total of 18 placebo-controlled trials examining acute efficacy met our criteria. The combined sample size in these studies was 2252. The mean sample size was 51 (range 20-728) and mean trial duration was 7 weeks. A total of 12 trials examined tricyclic antidepressants (TCAs), five trials examined selective serotonin reuptake inhibitors (SSRIs), two trials examined bupropion, and one trial examined mirtazapine. There were no published trials of venlafaxine or nefazodone. In all, 71.5% of trials reported significantly greater efficacy with drug than placebo. In conclusions, there is a paucity of published controlled antidepressant trials in the elderly. Most published studies examine small sample sizes and do not include common comorbid conditions. Efficacy studies examining relapse prevention are lacking. Large placebo response rates, lack of controlled head to head comparisons, and other methodological design differences make crosstrial comparisons difficult. Large simple studies are urgently needed to address the unmet needs for data on safety and efficacy of antidepressants in this population. PMID- 15340392 TI - Prefrontal neuropsychological predictors of treatment remission in late-life depression. AB - Recent studies suggest that neuropsychological measures involving the prefrontal cortex are associated with treatment remission in late-life depression. To further explore this issue, we studied the neuropsychological performance of 110 depressed individuals aged 60 years and over who are participating in an ongoing pharmacologic treatment study. Participants were clinically depressed at entry to the study as rated by the Montgomery-Asberg Depression Rating Scale (MADRS > or = 15), at which time they also completed a neuropsychological assessment that included measures of prefrontal/executive functions. A geriatric psychiatrist treating the participant using a standardized pharmacologic treatment algorithm evaluated the participant at baseline and 3-month follow-up, completing a MADRS at both visits. Using logistic discriminative procedures to predict depression remission at 3 months while controlling for age, gender, education, ethnicity, and baseline MADRS severity, we found that perseverative responses during verbal initiation tasks significantly predicted remission status (MADRS < 7). This finding is consistent with previous single-agent treatment studies suggesting a relationship between prefrontal neuropsychological function and treatment response in late-life depression. The current results, however, appear to differentiate verbal perseveration from verbal initiation as the cognitive process that is most associated with poor treatment response. By extension, we suggest that orbitofrontal prefrontal cortex may play a role in sustaining perseverative processing in geriatric depression. PMID- 15340393 TI - Executive dysfunction, heart disease burden, and remission of geriatric depression. AB - This study investigated the relationship of executive impairment and heart disease burden to remission of major depression among elderly patients. A total of 112 elderly subjects suffering from major depression received treatment with citalopram at a target daily dose of 40 mg for 8 weeks. Diagnosis was assigned using the Research Diagnostic Criteria and the DSM-IV Criteria after an interview with the Schedule for Affective Disorders and Schizophrenia. Executive dysfunction was assessed with the Initiation/Perseveration subscale of the Dementia Rating Scale (DRS) and the Color-Word Stroop test. Medical burden, including heart disease burden, was rated with the Cumulative Illness Rating Scale, and disability with Philadelphia Multilevel Instrument. Both abnormal initiation/perseveration and abnormal Stroop scores were associated with low remission rates of geriatric depression. Similarly, heart disease burden and baseline severity of depression also predicted low remission rates. The relationship of heart disease burden to remission was not mediated by executive dysfunction. Impairment in other DRS cognitive domains, disability, medical burden unrelated to heart disease did not significantly influence the outcome of depression in this sample. Executive dysfunction and heart disease burden constitute independent vulnerability factors that increase the risk for chronicity of geriatric depression. The findings of this study provide the rationale for investigation of the role of specific frontostriatal-limbic pathways in predisposing to geriatric depression or worsening its course. PMID- 15340394 TI - Lost in clinical translation. PMID- 15340395 TI - Edible vaccines not ready for main course. PMID- 15340396 TI - High-flying patents get their wings clipped in Europe. PMID- 15340397 TI - California dreaming about 'ill-conceived' stem cell plan. PMID- 15340398 TI - 'Fast-track' drug approvals hit speed bumps in Japan. PMID- 15340400 TI - Soaring drug prices send experts scrambling for a fix. PMID- 15340401 TI - NIH lab shutdown raises concerns about US prion research. PMID- 15340402 TI - WHO pulls Indian generics off AIDS drugs list. PMID- 15340403 TI - Malik Peiris. PMID- 15340404 TI - Cancer immunotherapy: breaking the barriers to harvest the crop. AB - Successful translation of modern molecular immunology into effective cancer immunotherapy is threatened by regulatory barriers and challenges to the development of novel agents and combinatorial strategies through effective public private partnerships. For its promise to be fully realized, both the National Cancer Institute and Food and Drug Administration must take active steps to help academic investigators and companies jointly navigate the pathways from laboratory to clinic. PMID- 15340407 TI - Autoimmune hypothyroidism: T cells caught in the act. PMID- 15340408 TI - Aneurysms: leukotrienes weaken aorta from the outside. PMID- 15340409 TI - Epithelial cells pay a Toll for protection. PMID- 15340410 TI - Fatal attraction: tumors beckon regulatory T cells. PMID- 15340411 TI - When good cholesterol goes bad. PMID- 15340412 TI - Mutated matrix gene mars macula. PMID- 15340413 TI - Lung immunity: necessity is the mother of induction. PMID- 15340414 TI - Maiming mitochondria in familial ALS. PMID- 15340416 TI - Cancer immunotherapy: moving beyond current vaccines. AB - Great progress has been made in the field of tumor immunology in the past decade, but optimism about the clinical application of currently available cancer vaccine approaches is based more on surrogate endpoints than on clinical tumor regression. In our cancer vaccine trials of 440 patients, the objective response rate was low (2.6%), and comparable to the results obtained by others. We consider here results in cancer vaccine trials and highlight alternate strategies that mediate cancer regression in preclinical and clinical models. PMID- 15340419 TI - Dividing the cloning divide. PMID- 15340420 TI - Estimating rates of alternative splicing in mammals and invertebrates. PMID- 15340423 TI - The knockout mouse project. AB - Mouse knockout technology provides a powerful means of elucidating gene function in vivo, and a publicly available genome-wide collection of mouse knockouts would be significantly enabling for biomedical discovery. To date, published knockouts exist for only about 10% of mouse genes. Furthermore, many of these are limited in utility because they have not been made or phenotyped in standardized ways, and many are not freely available to researchers. It is time to harness new technologies and efficiencies of production to mount a high-throughput international effort to produce and phenotype knockouts for all mouse genes, and place these resources into the public domain. PMID- 15340424 TI - The European dimension for the mouse genome mutagenesis program. AB - The European Mouse Mutagenesis Consortium is the European initiative contributing to the international effort on functional annotation of the mouse genome. Its objectives are to establish and integrate mutagenesis platforms, gene expression resources, phenotyping units, storage and distribution centers and bioinformatics resources. The combined efforts will accelerate our understanding of gene function and of human health and disease. PMID- 15340426 TI - As normal as normal can be? AB - Two papers report that large-scale copy-number variations, ranging in size from 100 kb to 2 Mb, are distributed widely throughout the human genome, and that a high proportion of them encompass known genes. This unexpected level of genome variation has implications for our view of human genetic diversity and phenotypic variation. PMID- 15340427 TI - Cancer chromosomes in crisis. AB - The benign-to-malignant transition in human breast cancer is associated with a marked increase in chromosomal aberrations. A new study suggests that telomere dysfunction and its associated bridge-fusion-breakage cycles may drive this episodic instability, thereby providing aspiring cancer cells with the multiple genetic aberrations needed to achieve a fully malignant state. PMID- 15340428 TI - TIMP3 checks inflammation. AB - Mice deficient in the metalloprotease inhibitor TIMP3, which inhibits the tumor necrosis factor alpha (TNF-alpha)-converting enzyme (TACE, also called ADAM17), have elevated levels of TNF and severe inflammation in the liver. This result confirms the physiological importance of the soluble form of TNF and identifies TIMP3 as a crucial regulator of this inflammatory cytokine. PMID- 15340429 TI - The G-netics of dark skin. AB - Several mutant strains of mice have dark skin pigmentation due to an aberrant accumulation of pigment-producing melanocytes in the dermal layer of the skin. A new study shows that three such strains carry activating mutations in the genes encoding the G-protein subunits Galphaq or Galpha11, resulting in more pigment cell precursors and an excess of dermally retained pigment cells at birth. PMID- 15340430 TI - From mRNA to tumor suppressor. AB - The combination of inhibition of RNA degradation and comparative genomic scanning is a powerful new method for detecting gene disruptions. The utility of the method is well-illustrated by a series of observations linking the ephrin receptor EPHB2 to prostate cancer. PMID- 15340433 TI - Towards sound epistemological foundations of statistical methods for high dimensional biology. AB - A sound epistemological foundation for biological inquiry comes, in part, from application of valid statistical procedures. This tenet is widely appreciated by scientists studying the new realm of high-dimensional biology, or 'omic' research, which involves multiplicity at unprecedented scales. Many papers aimed at the high-dimensional biology community describe the development or application of statistical techniques. The validity of many of these is questionable, and a shared understanding about the epistemological foundations of the statistical methods themselves seems to be lacking. Here we offer a framework in which the epistemological foundation of proposed statistical methods can be evaluated. PMID- 15340437 TI - Germany's 'junior professor' fails to germinate. PMID- 15340438 TI - Complex kinase requirements for EPEC pedestal formation. PMID- 15340442 TI - Francis Crick (8th June 1916 - 28th July 2004). PMID- 15340443 TI - A new checkpoint takes shape. PMID- 15340444 TI - Stretching cell fate. PMID- 15340445 TI - An Apollon vista of death and destruction. PMID- 15340446 TI - Spindle checkpoint protein links Rb pathway to aneuploidy. PMID- 15340447 TI - Unravelling Ras signals in cardiovascular disease. PMID- 15340448 TI - Nuclear reprogramming: a key to stem cell function in regenerative medicine. AB - The goal of regenerative medicine is to restore form and function to damaged tissues. One potential therapeutic approach involves the use of autologous cells derived from the bone marrow (bone marrow-derived cells, BMDCs). Advances in nuclear transplantation, experimental heterokaryon formation and the observed plasticity of gene expression and phenotype reported in multiple phyla provide evidence for nuclear plasticity. Recent observations have extended these findings to show that endogenous cells within the bone marrow have the capacity to incorporate into defective tissues and be reprogrammed. Irrespective of the mechanism, the potential for new gene expression patterns by BMDCs in recipient tissues holds promise for developing cellular therapies for both proliferative and post-mitotic tissues. PMID- 15340449 TI - Cell of the month: waves of glomerular podocytes. PMID- 15340450 TI - Homologue recognition during meiosis is associated with a change in chromatin conformation. AB - During meiosis, homologous chromosomes are sorted into pairs and are then intimately aligned, or synapsed, along their lengths while a proteinaceous structure, the synaptonemal complex, is assembled between them. However, little is known about how chromosomes first recognise each other. Here we show, by comparing the behaviour of wild-type wheat and wheat mutant for Ph1 (a suppressor of homeologous chromosome pairing), that when chromosomes recognise a partner to pair with, a conformational change to the chromatin is triggered in both partners that is followed by their intimate alignment [corrected]. Thus, a conformational change in the chromosomes at the onset of meiosis can be correlated directly with recognition. PMID- 15340451 TI - Orphans at the window. PMID- 15340452 TI - Cash-strapped biotechs find financing alternative. PMID- 15340453 TI - As products enter clinic...scientists shown the door. PMID- 15340454 TI - Russia becomes attractive as a source of IP for biotechs. PMID- 15340455 TI - EU to review rare disease drugs market exclusivity. PMID- 15340456 TI - NAS issues mixed message on unintended effects of GM food. PMID- 15340457 TI - France to focus on a few biotech clusters. PMID- 15340459 TI - Profile: Jim Greenwood. AB - A fervent supporter of embryonic stem cell research and a dogged opponent of corporate wrongdoers, US congressman Jim Greenwood is stepping out of politics to head biotech's biggest industry organization. PMID- 15340460 TI - Biotech R&D still reeling. PMID- 15340461 TI - The faking of champions. PMID- 15340462 TI - Coming soon: a global grid for cancer research. PMID- 15340463 TI - Developing market. PMID- 15340464 TI - The 802 million dollars fallacy. PMID- 15340465 TI - 'Informative' horizontal gene transfer. PMID- 15340466 TI - Hydrocarbons and renewable energy. PMID- 15340467 TI - AIDS, aneuploidy and oncogenes. PMID- 15340470 TI - Genomic islands in Rhodopseudomonas palustris. PMID- 15340471 TI - Keeping dry and crossing membranes. PMID- 15340472 TI - Does biotech M&A benefit investors? PMID- 15340473 TI - Goodbye Columbus! New NRDOs forego discovery. PMID- 15340474 TI - Phosphoproteomics finds its timing. PMID- 15340475 TI - Beta-cell precursors--a work in progress. PMID- 15340476 TI - Immunotherapy dispenses with tumor antigens. PMID- 15340477 TI - Finding enzymatic gold on silver surfaces. PMID- 15340478 TI - High-throughput biochemistry heats up. PMID- 15340480 TI - Problems in monitoring horizontal gene transfer in field trials of transgenic plants. AB - Transgenic crops are approved for release in some countries, while many more countries are wrestling with the issue of how to conduct risk assessments. Controls on field trials often include monitoring of horizontal gene transfer (HGT) from crops to surrounding soil microorganisms. Our analysis of antibiotic resistant bacteria and of the sensitivity of current techniques for monitoring HGT from transgenic plants to soil microorganisms has two major implications for field trial assessments of transgenic crops: first, HGT from transgenic plants to microbes could still have an environmental impact at a frequency approximately a trillion times lower than the current risk assessment literature estimates the frequency to be; and second, current methods of environmental sampling to capture genes or traits in a recombinant are too insensitive for monitoring evolution by HGT. A model for HGT involving iterative short-patch events explains how HGT can occur at high frequencies but be detected at extremely low frequencies. PMID- 15340481 TI - Monitoring and modeling horizontal gene transfer. AB - Monitoring efforts have failed to identify horizontal gene transfer (HGT) events occurring from transgenic plants into bacterial communities in soil or intestinal environments. The lack of such observations is frequently cited in biosafety literature and by regulatory risk assessment. Our analysis of the sensitivity of current monitoring efforts shows that studies to date have examined potential HGT events occurring in less than 2 g of sample material, when combined. Moreover, a population genetic model predicts that rare bacterial transformants acquiring transgenes require years of growth to out-compete wild-type bacteria. Time of sampling is there-fore crucial to the useful implementation of monitoring. A population genetic approach is advocated for elucidating the necessary sample sizes and times of sampling for monitoring HGT into large bacterial populations. Major changes in current monitoring approaches are needed, including explicit consideration of the population size of exposed bacteria, the bacterial generation time, the strength of selection acting on the transgene-carrying bacteria, and the sample size necessary to verify or falsify the HGT hypotheses tested. PMID- 15340482 TI - The exclusion of diagnostic methods from patentability by the EPC: a case for review? PMID- 15340486 TI - What is Bayesian statistics? PMID- 15340487 TI - Funding high-throughput data sharing. PMID- 15340488 TI - Strategic HR for today's biotechnology companies. PMID- 15340489 TI - Submission of microarray data to public repositories. PMID- 15340490 TI - Transcriptional control in the segmentation gene network of Drosophila. AB - The segmentation gene network of Drosophila consists of maternal and zygotic factors that generate, by transcriptional (cross-) regulation, expression patterns of increasing complexity along the anterior-posterior axis of the embryo. Using known binding site information for maternal and zygotic gap transcription factors, the computer algorithm Ahab recovers known segmentation control elements (modules) with excellent success and predicts many novel modules within the network and genome-wide. We show that novel module predictions are highly enriched in the network and typically clustered proximal to the promoter, not only upstream, but also in intronic space and downstream. When placed upstream of a reporter gene, they consistently drive patterned blastoderm expression, in most cases faithfully producing one or more pattern elements of the endogenous gene. Moreover, we demonstrate for the entire set of known and newly validated modules that Ahab's prediction of binding sites correlates well with the expression patterns produced by the modules, revealing basic rules governing their composition. Specifically, we show that maternal factors consistently act as activators and that gap factors act as repressors, except for the bimodal factor Hunchback. Our data suggest a simple context-dependent rule for its switch from repressive to activating function. Overall, the composition of modules appears well fitted to the spatiotemporal distribution of their positive and negative input factors. Finally, by comparing Ahab predictions with different categories of transcription factor input, we confirm the global regulatory structure of the segmentation gene network, but find odd skipped behaving like a primary pair-rule gene. The study expands our knowledge of the segmentation gene network by increasing the number of experimentally tested modules by 50%. For the first time, the entire set of validated modules is analyzed for binding site composition under a uniform set of criteria, permitting the definition of basic composition rules. The study demonstrates that computational methods are a powerful complement to experimental approaches in the analysis of transcription networks. PMID- 15340491 TI - Single nucleotide polymorphism-based validation of exonic splicing enhancers. AB - Because deleterious alleles arising from mutation are filtered by natural selection, mutations that create such alleles will be underrepresented in the set of common genetic variation existing in a population at any given time. Here, we describe an approach based on this idea called VERIFY (variant elimination reinforces functionality), which can be used to assess the extent of natural selection acting on an oligonucleotide motif or set of motifs predicted to have biological activity. As an application of this approach, we analyzed a set of 238 hexanucleotides previously predicted to have exonic splicing enhancer (ESE) activity in human exons using the relative enhancer and silencer classification by unanimous enrichment (RESCUE)-ESE method. Aligning the single nucleotide polymorphisms (SNPs) from the public human SNP database to the chimpanzee genome allowed inference of the direction of the mutations that created present-day SNPs. Analyzing the set of SNPs that overlap RESCUE-ESE hexamers, we conclude that nearly one-fifth of the mutations that disrupt predicted ESEs have been eliminated by natural selection (odds ratio = 0.82 +/- 0.05). This selection is strongest for the predicted ESEs that are located near splice sites. Our results demonstrate a novel approach for quantifying the extent of natural selection acting on candidate functional motifs and also suggest certain features of mutations/SNPs, such as proximity to the splice site and disruption or alteration of predicted ESEs, that should be useful in identifying variants that might cause a biological phenotype. PMID- 15340492 TI - Monomethyl branched-chain fatty acids play an essential role in Caenorhabditis elegans development. AB - Monomethyl branched-chain fatty acids (mmBCFAs) are commonly found in many organisms from bacteria to mammals. In humans, they have been detected in skin, brain, blood, and cancer cells. Despite a broad distribution, mmBCFAs remain exotic in eukaryotes, where their origin and physiological roles are not understood. Here we report our study of the function and regulation of mmBCFAs in Caenorhabditis elegans, combining genetics, gas chromatography, and DNA microarray analysis. We show that C. elegans synthesizes mmBCFAs de novo and utilizes the long-chain fatty acid elongation enzymes ELO-5 and ELO-6 to produce two mmBCFAs, C15ISO and C17ISO. These mmBCFAs are essential for C. elegans growth and development, as suppression of their biosynthesis results in a growth arrest at the first larval stage. The arrest is reversible and can be overcome by feeding the arrested animals with mmBCFA supplements. We show not only that the levels of C15ISO and C17ISO affect the expression of several genes, but also that the activities of some of these genes affect biosynthesis of mmBCFAs, suggesting a potential feedback regulation. One of the genes, lpd-1, encodes a homolog of a mammalian sterol regulatory element-binding protein (SREBP 1c). We present results suggesting that elo-5 and elo-6 may be transcriptional targets of LPD-1. This study exposes unexpected and crucial physiological functions of C15ISO and C17ISO in C. elegans and suggests a potentially important role for mmBCFAs in other eukaryotes. PMID- 15340493 TI - [Development of recombinant human-derived IgG antibody against HBsAg]. AB - BACKGROUND: To investigate preparation of human-derived anti-HBs IgG antibody. METHODS: mRNA was extracted from lymphocytes of a volunteer with high level antibody to HbsAg. Fab phage display library was constructed. Purified HBsAg was used to screen the Fab. Eukaryotic expression vector was constructed and transfected into sf9 cells, then the supernatant was purified. The expression of anti-HBs human IgG was tested with blocking ELISA and immunofluorescent test. RESULTS: A whole human anti-HBs IgG molecule was obtained, the antibody reacts with HBsAg "a" determinant. CONCLUSION: It can be concluded preliminarily that this antibody has neutralizing characteristic against HbsAg. PMID- 15340494 TI - [Distribution and timing of antibody to SARS-CoV in SARS cases of transmission chain or non-transmission chain]. AB - BACKGROUND: To find out the timing of serologic responses after illness onset and distribution of IgG antibody to SARS-CoV in SARS cases of transmission chain or non-transmission chain. METHODS: The IgG and IgM antibodies to SARS-CoV were tested by indirect ELISA in serum samples from 301 clinically diagnosed SARS cases. RESULTS: Totally 158 SARS cases were involved in 15 chains of transmission. The positive rates of SARS-CoV IgG in those chains were 85.70% 100.00% and the overall rate was 94.30% (149/158). The chain of transmission could spread to four generations, but the SARS cases were reduced with increase of generations. There was no significant difference among positive rates of SARS CoV IgG for generations, Chi square=5.11, P greater than 0.05. The positive rate of SARS-CoV IgG in cases who were not in chain of transmission was 12.59%(18/143) which was statistically significantly different from that of cases in chain of transmission, Chi square=199.64, P less than 0.001. During days 0-7,8-14,15-21,22 30 after onset, the cumulated positive rate of SARS-CoV IgG was 16.67%, 40.00%, 70.00% and 93.10%, respectively, then was kept at the level above 90% and lasted for 217 days. The cumulated positive rate of SARS-CoV IgM during days 0-7 after onset was the same to that of IgG. During days 8-14, 55.17% of cases had seroconversion for IgM which reached a peak (86.96%) during days 21-30. Then the rate rapidly declined. CONCLUSION: More than 94% of cases with SARS could produce IgG antibody when they were infected by SARS-CoV. Detecting SARS-CoV IgG could provide a diagnostic evidence for case confirmation. SARS-CoV IgG appeared as early as 7 days after onset and reached the peak at about weeks 4. Then the high rate of antibody was maintained for more than 6 months. PMID- 15340495 TI - [Construction and immune potency of recombinant adenovirus containing codon modified HIV-1 gp120]. AB - BACKGROUND: To construct replication-deficient recombinant adenovirus expressing wild and codon-modified HIV-1 gp120. METHODS: The viral codons were changed to the codon usage of highly expressed mammal gene, the resulting modified gp120 gene was synthesized. The wild and modified gp120 genes were cloned into shuttle vector pShuttle-CMV respectively, and then the constructed plasmids containing gp120 gene was cotransformed with the backbone vector pADeasy-1 into E.coli BJ5183. Transfection of the recombinant AdEasy plasmid into 293 cells was performed to obtain recombinant adenoviruses. The mice were immunized with the recombinant adenoviruses. Their immunogenicity was evaluated by testing antibody and CTL levels of immunized mice. RESULTS: Two strains of recombinant adenovirus expressing wild and codon-modified HIV-1 gp120 were obtained. The protein expressing level of the recombinant adenoviruses containing modified genes was much higher than that containing wild genes. The mice immunized with recombinant adenoviruses elicited HIV-1 specific antibody and CTL response. The rAd-mod gp120 group was better than the rAd-wt gp120 group. CONCLUSION: Replication-deficient recombinant adenovirus expressing HIV-1 gp120 can elicit HIV-1 specific humoral and cellular response, the codon-modified recombinant virus was more efficient than the native. PMID- 15340496 TI - [Influence of ultrasonic processing on the aggregation of PrP-Sc in the brain extracts of the scrapie-infected hamsters]. AB - BACKGROUND: To evaluate the influence ultrasonic processing on the aggregation of PrP(Sc) in the brain extracts prepared from the scrapie-infected hamsters, and to seek for the way to prepare lower molecular PrP(Sc) polymer. METHODS: The extracts of infected brains were prepared with a lysis solution, and treated with ultrasonics at various conditions during the different phases. The distribution and aggregation state of PrP(Sc) were analyzed by proteinase K treated. Western blot, and afterwards, quantitatively calculated with a commercially supplied software Image Totaltech. RESULTS: The amount of PrP(Sc) in the supernatant of brain homogenates was 1.29-to 1.58-fold increased with appropriate sonication (15 s for 30 times). At the same conditions of ultrasound, the PrP amount in the supernatant prepared from the scrapie-infected hamster brain was significantly increased, whereas that prepared from healthy animal used as normal control showed little change. Comparative analyses of PrP(Sc) pellets prepared by high speed centrifugation revealed that about 90% PrP(Sc) released into supernatant after ultrasound processing. CONCLUSION: Appropriate sonication of homogenate of scrapie-infected brain increases the extracted amount of PrP(Sc), being favorable to laboratory diagnosis. Larger molecular PrP(Sc) aggregates can be crashed by ultrasonic processing, engendering lower molecular PrP(Sc) polymers. PMID- 15340497 TI - [Construction of full-length complementary DNA of hepatitis C virus genome from an HCV infected patient]. AB - BACKGROUND: To construct the full-length complementary DNA of HCV genome from an HCV infected patient. METHODS: Four HCV gene fragments (1.6, 3.5, 2.4 kb and 2.6 kb) were amplified by RT-PCR from serum of a Chinese patient and fused and connected together to produce a 9.2 kb subgenomic fragment, which was further cloned into a cassette vector with fixed 5-prime and 3-prime termini of HCV to make the full-length cDNA. The cDNA heterogeneity was analyzed by comparing the sequences of 4 isolated HVR1 regions. The prokaryotic expressed Core, NS3 protease, NS3 helicase were detected for their specific reactivities with patient serum by Western blot analysis. And the protease activity of NS3 was evaluated in a cell-based NS3/4A-SEAP expression system. RESULTS: The cDNA covered the near full-length of HCV genome from the patient's serum. The difference among HVR1 regions indicated no selection of HCV variants during RT-PCR and the quasi species characteristic of the amplified cDNA. The prokaryotic expressed viral proteins could be identified by patient serum. In the NS3/4A-SEAP system, NS3 could cleave the 4A-4B site between NS4A and SEAP proteins and resulted in the secretion of SEAP in culture media. CONCLUSION: These results suggest that the cloned HCV cDNA encodes a complete and functional open reading frame and will be useful for further construction of infectious cDNA clone. PMID- 15340498 TI - [Prevalence of anti-HEV among swine, sheep and chickens]. AB - BACKGROUND: To investigate the prevalence of anti-HEV among swine, sheep and chickens. METHODS: Totally 498 sera of swine, sheep and chickens collected from Xingjiang, Guangxi, Guangdong, Beijing and Hebei were detected for the anti-HEV by an enzyme linked immunoassay. RESULTS: The anti-HEV positive rate of swine was 67.53%(104/154), in pigs between 4-5 months of age the rate was 100.00%(9/9) from Xingjiang. The rate in pigs under 3 months of age from Guangxi was 36.00%(9/25) and in pigs older than six months of age was 71.67% (86/120), respectively. The 108 sera of sheep collected from Xingjiang were all negative. The positive rate of chickens was only 1.27% (3/236). The anti-HEV prevalence rates of chickens from Luoding, Shenzhen, Liuzhou, Beijing and Hebei were 4.00%, 1.49%, 1.49%, 0, 0 respectively. CONCLUSION: HEV infection does exist among swine and chickens. The anti-HEV prevalence of swine was the highest among domestic animals. The role of swine and chickens in transmission of HEV needs to be further studied. PMID- 15340499 TI - [Effect of duration and temperature of sample preservation on the result of peripheral blood CD4+ and CD8+ T lymphocyte count in HIV/AIDS patients]. AB - BACKGROUND: By analyzing the CD4+ and CD8+ T lymphocyte count of whole blood from HIV/AIDS patients, which were stored at different temperatures for various durations, the authors studied the ideal preserving condition for whole blood and processed, in a purpose of guaranteeing the accuracy of clinical testing of CD4+ and CD8+ T lymphocyte count. METHODS: Blood from 34 HIV carriers/AIDS patients, were kept at 4 degrees C for 2, 24, 48, or 72 h, and tested for CD4+ and CD8+ T lymphocyte count using cytometric analysis. Part of the blood was processed, and kept at degrees C or room temperature for 2, 24, 48, or 72 h, then tested for CD4+ and CD8+ T lymphocyte count. The results were compared statistically in parallel. RESULTS: Whole blood and processed samples preserved at degrees C showed no statistical difference in CD4+ T lymphocyte count among different preserving durations (P greater than 0.05), but CD8+ T lymphocyte counts were significantly different at 72 h (P less than 0.05). Processed samples at 72 h were significantly different in CD4+ T lymphocyte count(P less than 0.05), and significantly different in CD8+ T lymphocyte count at 24 h (P less than 0.05). At room temperature, samples at different duration were not significantly different in CD4+ T lymphocyte count, but significantly different in CD8+ T lymphocyte count at 48 and 72 h (P less than 0.05). CONCLUSION: There were stable results for performing analysis of the CD4+ and CD8+ T lymphocyte count of the anticoagulated blood within 48 h. At room temperature, there were stable results for performing the analysis of CD4+ and CD8+ T lymphocyte count of processed samples within 24 h. Between 24 h and 48 h, although CD4+ count was stable, CD8+ count showed significant changes, so the ratio of CD4 to CD8 changed accordingly. PMID- 15340500 TI - [Clinical outcomes of women with transfusion-associated hepatitis C after 10-15 years follow-up]. AB - BACKGROUND: To investigate natural history of hepatitis C virus (HCV) infection and related factors in China. METHODS: Totally 41 women infected with HCV 10-15 years ago were evaluated in this study. Their clinical manifestations were recorded and sera tested for alamine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (gamma-GT), alkaline phosphatase (ALP), total bilirubin (TBIL), direct bilirubin (DBIL), total protein (TP), albumin (ALB), and alpha-fetoprotein (alpha-FP), as well as virus markers. The B-type ultrasound examination was performed on the liver, gall bladder, spleen and pancreas. For virus markers, HCV RNA was detected with fluorogenic quantitative polymerase-chain-reaction, HBsAg and anti-HCV antibody, with enzyme linked immunosorbent assay (ELISA). RESULTS: Ten to 15 years after infection, the infection was cleared spontaneously in 8 cases (19.51%); 73%(30/41) cases had signs at present, 32% (13/41) cases had mild ALT and/or AST elevation. On ultrasound examination, 83%(34/41) had mild degree 17%(7/41) moderate degree changes of the viral hepatitis. CONCLUSION: Women with HCV infection for 13 +/- 1 (10-15) years after transfusion had slight hepatic inflammation. A part of them had cleared the virus spontaneously. PMID- 15340501 TI - [Difference and significance of T-lymphocyte subsets in differential diagnosis between severe acute respiratory syndrome and common atypical pneumonia]. AB - BACKGROUND: To clarify the difference and significance of T-lymphocyte subsets in differential diagnosis between severe acute respiratory syndrome SARS) and common atypical pneumonia. METHODS: Totally 100 patients hospitalized in Beijing Ditan Hospital since March to June 2003 with clinical diagnosis of SARS were involved in this study. These patients courses of disease were over 3 weeks. These patients were divided into two groups, SARS group and common atypical pneumonia group (non-SARS group). The counts of CD3+, CD4+ and CD8+ T-lymphocyte of two groups were systematically recorded and analyzed. RESULTS: Sixty-five of the patients were confirmed to have common type of SARS, including 26 males and 39 females, 50 cases received methylprednisolone treatment. Thirty-five cases had common atypical pneumonia (non-SARS), 21 were males while 14 were females, 20 cases received methylprednisolone treatment. All the cases of two groups were cured in the end. The SARS patients T-lymphocyte counts decreased first and then increased. Before 15 days of disease course, mean CD3+, CD4+, CD8+ T-lymphocyte counts of SARS patients were decreased apparently (694+/-568/microl, 441+/ 356/microl, 309+/-462/microl). After 15th day of disease course, the counts gradually returned to normal CD3+, CD4+, CD8+ T-lymphocyte counts of non-SARS patients were normal. Compared with patients of the same group who were not treated with glucocorticoids, T-lymphocyte counts of non-SARS patients treated with glucocorticoids had no obvious difference. But glucocorticoids had some effect on SARS patients recovery of cellular immune function, i.e., it delayed the recovery by about 6 days. CONCLUSION: With or without treatment with glucocorticoids,the lowered CD3+, CD4+, CD8+ T-lymphocyte counts in the early stage are of very important significance in differential diagnosis between severe acute respiratory syndrome and common atypical pneumonia. PMID- 15340502 TI - [Association of T cell subsets with clinical status and hepatic pathology in children with chronic hepatitis B virus infection]. AB - BACKGROUND: To study characteristics of peripheral T cell subsets in 94 children with chronic hepatitis B and to elucidate its relationships with clinical status and hepatic pathology. METHODS: Peripheral T cell subsets were detected using flow cytometric analysis with specific monoclonal antibodies staining in 94 patients with HBV infection. The authors simultaneously detected their serum ALT, markers of HBV infection and examined liver biopsy material for pathological changes. RESULTS: In patients with serious liver lesion, the ratio of CD4+/CD8+ cells was significantly higher than those with mild lesion (1.41+/-0.54 vs 1.08+/ 0.35, P less than 0.05), which seemed to be associated with the various liver lesions among the patients. In female cases, the levels of CD4+ T cells and the ratio of CD4+/CD8+ T cells were higher than their counterpart in male cases (33.1+/-5.39 vs 28.8+/-6.28, 1.28+/-0.32 vs 1.02+/-0.36, P less than 0.05), but the level of CD8+ T cells was lower than those in males (26.79+/-4.66 vs 30.51+/ 7.17, P less than 0.05). There was no obvious correlation between T cell subsets and circulating HBV viral load, the size of spleen among the HBV-infected children. CONCLUSION: The characteristics of peripheral T cell subsets probably suggests the immune disorder occurred in these children with hepatitis B compared with healthy controls and its mechanism needs further investigation. PMID- 15340503 TI - [Study on anti-HBV effects of genetically engineered replication-defective hepatitis B virus expressing dominant negative mutants of core protein]. AB - BACKGROUND: To explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular expression of dominant negative mutants of core protein. METHODS: Two kinds of full length mutant HBV genome, which express Core-partial P and Core-S fusion protein, were transfected into HepG 2.2.15 cell lines. Positive clones were selected and mixed in respective groups with hygromycin in the culture medium. HBsAg and HBeAg, which exist in the culture medium, were tested by ELISA and intracellular HBc related HBV DNA was examined by dot blot hybridization. The existence of recombinant HBV virion in the culture medium was examined by PCR. RESULTS: The mean inhibitory rates of HBsAg were 2.74+/-3.83%, 40.08+/-2.05% (P less than 0.01) and 52.94+/-1.93% (P less than 0.01) in group 2.2.15-pMEP4, 2.2.15-CP and 2.2.15-CS, respectively. The mean inhibitory rates of HBeAg were 4.46+/-4.25%, 52.86+/-1.32% (P less than 0.01) and 41.60+/-1.65% (P less than 0.01), respectively. The inhibitory rates of HBc related HBV DNA were 15.3%, 82.0% and 67.2%, respectively. Recombinant HBV virion was detectable in the culture medium of only group 2.2.15-CP. CONCLUSION: Dominant negative mutants of core protein can efficiently suppress wt-HBV replication and the expressions of HBV antigens. With the help of wild-type HBV, the recombinant HBV genome can form and secret HBV like particles, which provides evidence that the antiviral gene will be hepatotropic expression and the antiviral effects will be amplified. PMID- 15340504 TI - [Construction of replication-deficient recombinant adenovirus expressing gag polDelta and gp140TM genes of human immunodeficiency virus in mice]. AB - BACKGROUND: Construction of replication-deficient recombinant adenovirus expressing gag-pol and env genes of human immunodeficiency virus (HIV) in mice. METHODS: gag-polDelta and gp140TM genes were cloned into shuttle vector pAdTrack CMV respectively, and then the plasmids containing gag-polDelta or gp140TM gene were cotransformed with the backbone of adenovirus into E.coli BJ5183. Transfections of the recombinants were performed to obtain recombinant adenoviruses. Its immunogenicity was evaluated by testing antibody levels of mice primed with DNA vaccines and boosted with recombinant adenoviruses. RESULTS: The replication-deficient recombinant adenovirus could express Gp140TM, Gag P55 and P24 proteins correctly. The mice primed with DNA vaccines and boosted with recombinant adenoviruses elicited high titer of HIV-1-specific antibody compared with that inoculated with DNA vaccines only. CONCLUSION: Replication-deficient recombinant adenovirus expressing gag-polDelta and gp140TM can elicit high titer HIV-1-specific antibodies. PMID- 15340505 TI - [Polymorphism analysis of human cytomegalovirus UL148 gene in low passage clinical isolates]. AB - BACKGROUND: To investigate the polymorphism of human cytomegalovirus UL148 gene in low passage clinical isolates and to study the relationship between the polymorphism and different pathogenesis of congenital HCMV infection. METHODS: PCR was performed to amplify the entire HCMV UL148 gene region of 38 clinical isolates, which had been proven containing detectable HCMV-DNA by using FQ PCR.PCR amplification products were sequenced directly and the sequence data were analysed. RESULTS: Seventeen of 38 isolates were amplified successfully. By comparison with Toledo sequence, the length of UL148 ORFs in all 17 clinical isolates was similar to that of Toledo. Amino acid variability rate of UL148 protein was 0.3%-2.3%. There were additional or deleted sites of posttranslational modification of UL148 protein in all clinical isolates. CONCLUSION: All DNA and deduced amino acid sequences of UL148 gene shared great similarity among HCMV clinical strains regardless of their polymorphism. PMID- 15340506 TI - [Construction and expression of DNA-binding domain plasmid with hepatitis B virus e antigen in yeast double hybrid system]. AB - BACKGROUND: Using hepatitis B virus e antigen (HBeAg) gene to construct the DNA binding domain vector, which can express HBeAg in yeast cell, and can be used in yeast double hybrid as "bait plasmid" to look for the gene from the cDNA library, which expresses the protein that can interact with HBeAg. METHODS: PCR was performed to amplify the HBeAg gene from a sera of hepatitis B patient. The product of the amplification was inserted into T-vector and was verified by sequencing. Then it was inserted into the "bait" plasmid pGBKT7 after the digestion with the restricted endonuclease of EcoR I and Sal I. The plasmid was transformed into the yeast cell. PCR was used to verify whether the plasmid was transformed into yeast. The HBeAg protein expressed in the cell was confirmed by Western blot. Using nutrition selection assay to verify the constructed plasmid alone could not activate the reporter gene in the yeast cell. RESULTS: Sequenced and digested by two endonucleases, the recombined vectors pGBKT7-eAg produced anticipated fragment. PCR verified that there was HBeAg fragment in the yeast. Having assayed by Western blotting, it was shown that the yeast cell transformed with pGBKT7-eAg vector had positive signal which could not be seen in the control. Tested by the nutrition selection assay, the recombined vectors pGBKT7 eAg could not activate LacZ reporter gene in the yeast. CONCLUSION: DNA-binding domain plasmid was successfully constructed and could express HBeAg proteins in the yeast cell but could not activate transcription of LacZ reporter gene alone. The recombined plasmid can be used in yeast double hybrid. PMID- 15340507 TI - [Significance of detecting HBV-DNA by the fluorescence quantitative PCR]. AB - BACKGROUND: To study the correlativity between HBV-DNA and the markers of hepatitis B virus infection and different clinical types of hepatitis B. METHODS: The fluorescence quantitation (FQ) of HBV-DNA of 105 patients with hepatitis B was performed by PCR, and the correlativity between the fluorescence quantitation of HBV-DNA and the markers of hepatitis B virus and different clinical types of hepatitis B was analyzed. RESULTS: Ninety-seven percent of the patients were found HBsAg(+), HBeAg(+), HBcAb(+); 75% were HBsAg(+), HBeAb(+), HBcAb(+); 60% were HBsAg(+), HBcAb(+); 40% were HBsAg(+); in HBsAb(+), HBeAb(+), HBcAb(+) (or both HBsAb and HBcAb were positive) group the HBV DNA was undetectable. The analysis indicated that there was a significant difference among different groups (P less than 0.05).HBV-DNA was detected in 72.2% in acute hepatitis B group, in 75% of chronic hepatitis B group, and in 70% of cases of liver cirrhosis with hepatitis B group. The analysis indicated that there was no significant difference among the different clinical types of hepatitis (P greater than 0.05). CONCLUSION: The levels of viral replication were not correlated with different clinical types of hepatitis B; the concentration of HBV-DNA in serum was related to hepatitis B antigen. PMID- 15340508 TI - [Cellular ultrastructural changes of bone marrow of patients with hemorrhagic fever with renal syndrome]. AB - BACKGROUND: To observe cytopathogenic effect of Hantaan virus (HV) on cultured human bone marrow cells. METHODS: Light and transmission electron microscopy and direct immunofluorescent technique were applied to study cellular structure especially ultrastructural changes of bone marrow cells from patients with Hantaan virus infection. Bone marrow cells of one healthy volunteer were also studied as control. RESULTS: The antigen of HV was found in bone marrow cells of 20 of 27 HFRS patients by the aid of direct immunofluorescent technique. It was found that the granulocytes had the highest percentage of HV antigen positive cells (76%), followed by monocytes (65%), lymphocytes (40%), megakaryocytes (20%) and the lowest was found in erythrocytes (3.7%). The injury of cell membrane after infection with HV was significantly more severe than that in the control group under the light and electron microscopy. CONCLUSION: This study demonstrated that HV could attack human bone marrow cells and cause cytopathogenic effect on them. PMID- 15340509 TI - [Study on clinical isolates of human cytomegalovirus found in Urumqi by restriction fragment length polymorphism of PCR products]. AB - BACKGROUND: The study was designed to investigate the status of molecular epidemiology of HCMV in Urumqi through genetic comparison of clinical isolates. METHODS: DNA sequences of 2.0-2.6 kb were amplified by polymerase chain reaction from three relatively conservative gene regions (DNA polymerase, glycoproteins H, and major immediate-early antigen) of 28 clinical HCMV strains and then were analysed by restriction enzymes. RESULTS: The restriction patterns of the clinical isolates which did not have relation in epidemiology were greatly different, but the patterns of the clinical isolates related in epidemiology such as strains paired in mother and infant were quite similar. Of eight mother and infant pairs, from whom HCMV were isolated, four pairs showed identity of restriction profiles within each pair for all three amplified regions, four pairs showed differences between mother and infant. CONCLUSION: These results confirm the high degree of genetic variability among cytomegalovirus strains in Urumqi. Analysis of PCR-RFLP can indicate transmission of HCMV infection and facilitate its molecular epidemiologic studies. PMID- 15340510 TI - [Oxidative stress in patients with chronic hepatitis B]. AB - BACKGROUND: To study oxidative stress in patients with chronic hepatitis B. METHODS: Malondialdehyde (MDA), total anti-oxidative ability and ascorbic acid were measured as markers of oxidative stress in 30 patients with chronic hepatitis B, besides HBV DNA and ALT. RESULTS: MDA was significantly higher in patients with hepatitis B than the controls (P less than 0.05). Ascorbic acid was significantly higher in patients with normal ALT than the controls (P less than 0.01). MDA was significantly higher in patients with increased ALT than the controls and in patients with normal ALT. MDA was significantly positively correlated with ALT (r=0.61), and ascorbic acid was significantly negatively correlated with ALT (r=-0.64) in patients with hepatitis B. No significant relationship was found between HBV DNA and other indices of oxidative stress. No significant difference in total anti-oxidative ability was found among all groups. CONCLUSION: There was a disturbance between oxidative stress and anti oxidative ability in patients with chronic hepatitis B. In patients with increased ALT, oxidative stress became high. In patients with normal ALT, oxidative stress level was low. The indices of oxidative stress should be detected in patients with hepatitis B, in addition to HBV markers. PMID- 15340511 TI - [Apoptosis of peripheral blood cells of children with viral pneumonia]. AB - BACKGROUND: To study apoptosis of peripheral blood cells of children with viral pneumonia, explore immunopathogenesis and the possibility of immunotherapy of patients with viral pneumonia. METHODS: Fresh peripheral blood samples were collected from 28 patients with viral pneumonia and 24 healthy children were treated and run through the flow cytometry. The data were acquired using Cell Quest software and the percentage of live cells, viable apoptotic cells, non viable apoptotic cells and dead cells of neutrophils and lymphocytes were counted. The patients with viral pneumonia were hospitalized at our hospital. The average age of patients was 1.3 years; 24 healthy children were served as control group (age 1.8 years, on average). T-test and variance analysis by SPSS FOR WINDOWS 10.0 software was used for statistical analysis. RESULTS: The percentage of live neutrophils and lymphocytes in the acute stage and recovery stage in patients were significantly lower than that in control group (P < 0.01). The percentage of viable apoptotic neutrophils and lymphocytes in two stages in patients were significantly higher than that in control group (P < 0.05). Except for the percentage of live cells, non-viable apoptotic cells and dead lymphocytes, others had no difference between the patients and control groups. CONCLUSION: Apoptosis of neutrophils and lymphocytes of peripheral blood cells of children with viral pneumonia increased. Whereas the percentage of live cells decreased. Drugs that can accelerate apoptosis may be helpful in treatment of viral pneumonia. PMID- 15340512 TI - [A prospective study on short period antibiotic treatment of hepatic failure complicated with spontaneous bacterial peritonitis]. AB - BACKGROUND: To observe the effects of short-term antibiotic treatment in patients with hepatic failure and spontaneous bacterial peritonitis (SBP). METHODS: In this prospective study short-term antibiotic treatment was given to 67 cases diagnosed as hepatic failure with spontaneous bacterial peritonitis. Ceftriaxone 2 g, iv drip, q12h for 10 d or ofloxacin 0.2 g, iv drip, q12h for 10 d was given to the patients at random and the efficacy was evaluated on the basis of clinical symptoms, medical examination and ascites after 3, 7, 10 days of therapy. RESULTS: Seven cases (10.44%) were cured and 57 cases (85%) were improved after 3 days therapy, the total effective rate was 95.52%, but in 3 cases the therapy had no effect. The results of ascites bacterial culture and drug susceptibility test showed that one case had drug resistance to ceftriaxone and two cases had drug resistance to ofloxacin, so antibiotics were changed in time. After 7 days therapy, the results showed that 65 cases (97.01%) cured and 2 cases (2.99%) were improved, the total effective rate was 100%. When the therapy lasted for 10 days, all patients were cured. One patient had oral mucous membrane. Candida albicans infection after 3 days therapy; two cases got thrush and one patient got fungal intestinal infection after 7 days therapy; when the therapy lasted for 10 days, 4 cases had thrush and 2 cases had fungal infection of intestines and one patient had pulmonary fungal infection. CONCLUSION: The optimal period of antibiotic treatment of hepatic failure with SBP should be from 7 days to 10 days. PMID- 15340513 TI - [Producing human lactoferrin by high-density fermentation recombinant Pichia pastoris]. AB - BACKGROUND: To evaluate expression of human lactoferrin gene by high-density fermentation in recombinant Pichia pastoris on the premise of maintaining its biological activities. METHODS: The neutrophil was isolated from human peripheral blood and its total RNA was prepared. Full-length cDNA of human lactoferrin gene was then obtained by RT-PCR, cloned into expression vector pPIC 3.5 K and transformed into Pichia pastoris strain KM71. With two-layer filter method, the transformants with high-productivity of human lactoferrin were screened out into fed-batch high-density fermentation. And later, the physical, chemical and biological activities of fermentation product were detected preliminarily. RESULTS: The strain p3.5-k-7 with better productivity of human lactoferrin was screened out into fed-batch high-density fermentation. The fermentation lasted nearly for nine days, with A-600 of culture once above 260 and the highest productivity of human lactoferrin being 115 mg/L, 7.67 times the amount of that in shake flask cultivation. CONCLUSION: The authors successfully realized high density fermentation expression of human lactoferrin gene in recombinant Pichia pastoris. PMID- 15340514 TI - [Clinical research on the effect of Oxymatrine on serum cholinesterase]. AB - BACKGROUND: To investigate the effect of Oxymatrine (OM) on serum cholinesterase (ChE) during the treatment of viral hepatitis and the relationship between the change of ChE and the change of albumin (ALB), prothrombin activity (PTA) and other liver function tests. METHODS: A total of 98 patients with viral hepatitis were divided into four groups. Group A consisted of 31 patients and were treated with OM intravenous infusion; Group B consisted of 30 patients, treated with OM orally; Group C consisted of 7 patients and were treated with OM intramuscular injection while Group D consisted of 30 patients, and were not treated with OM. ChE, ALB, PTA, liver function, renal function, soluble complement receptor-1 (sCR1) and erythrocyte innate immune adhesion function (EIIAF) were regularly determined. RESULTS: ChE in Group A,B,C was dropped obviously during the treatment (P less than 0.001, less than 0.001, 0.023=. But there were no change in ALB, PTA, sCR1, EIIAF (P greater than 0.05), and remarkable improvement of ALT, AST, TBiL was seen during the treatment in Groups A, B, C. After the treatment with OM, the level of ChE recovered soon. CONCLUSION: Serum level of ChE significantly declined during the treatment of viral hepatitis with OM, but no change was found in ALB, PTA, sCR1, EIIAF while liver function tests showed better results. So the drop of ChE does not mean deprivation of patient's liver disease. PMID- 15340515 TI - [Construction of a genechip method for detection of hepatitis B virus DNA, basal core promotor and Pre-C mutants]. AB - BACKGROUND: To establish a genechip method for detection of hepatitis B virus (HBV) DNA, basal core promotor (BCP), and Pre-C mutants. METHODS: This study used two kinds of technology (PCR, oligochip), which can detect five mutant hotspots including nt 1 896, nt 1 899, nt 1 862, nt 1 764 and nt 1 762. The results of genechip method was verified by DNA sequencing. RESULTS: In detecting HBV DNA, the results of genechip were 100% consistent with those of DNA sequencing. In detecting HBV BCP and Pre-C mutants, 146 codons showed the same results using both methods, except for only 4 codons (P greater than 0.05). CONCLUSION: This convenient high throughput genechip method could detect several BCP and Pre-C mutant codons at the same time. These results suggest that genechip method has the same positive rate and specificity with DNA sequencing method. It has more advantages than the latter in detecting mixed mutants and therefore may be used in clinical practice. PMID- 15340516 TI - [Characterization of HA and NA genes of swine influenza A (H9N2) viruses]. AB - OBJECTIVE: To understand the origin of HA and NA genes of swine influenza A (H9N2) viruses isolated from pigs in the mainland of China and on basis of these to reveal the pathogenicity of them in pigs. METHODS: The target gene was amplified by PCR, the PCR product was ligated with PGEM-T Easy Vector (Promega company, USA) at 4 degrees, the recombined plasmid was transferred into DH-10 beta bacteria; positive colonies were selected and identified then digested with restriction enzyme. Afterwards,the nucleotide sequence was determined. Finally,phylogenetic analysis of the sequencing data was performed with MegAlign (Version 1.03) and Editseg (Version 3.69) softwares. RESULTS: Two strains of swine influenza A(H9N2) virus isolated in the mainland had an amino acid residue, leucine (L) at position 226 (H3 numbering) on HA protein molecule found in H9N2 viruses isolated either in pigs or humans previously; the amino acid sequence at HA connecting peptide of isolates possessed R-L-S-R, whereas the other H9N2 viruses with virulence in poultry had R-S-S-R at HA connecting peptide. The two pig H9N2 isolates shared the same three-amino-acids deletion in the NA stalk at 62.64 position found in A/Shaoguan/408/98 and A/Swine/Hong Kong/9/98, as well as A/Duck/Hong Kong /Y280/97(H9N2) viruses. The analysis of the phylogenetic tree indicated that the HA and NA genes of new isolates were closely related to those of A/Chicken/Hong Kong/G23/97 and A/Chicken/Hong Kong/G9/97 and A/Shaoguan/408/98 viruses, respectively. CONCLUSION: The HA and NA genes of swine influenza A(H9N2) viruses isolated in the mainland of China probably were derived from those of avian influenza A(H9N2) virus. The occurrence of substitution of amino acid sequence at HA connecting peptide, could result in the H9N2 virus from non pathogenic to pathogenic in pigs. However, avian influenza A(H9N2) virus had deletion in the stalk of the NA that resulted in host range transmission. Therefore they could infect pigs directly. PMID- 15340517 TI - [Expression of main neutralization antigen VP7 of human rotavirus serotype G2 and G3 by recombinant adenoviruses]. AB - OBJECTIVE: To express main neutralization antigen VP7 of human rotavirus serotype G2 and G3 by recombinant adenoviruses on the basis of previous investigation of the prevalence of rotavirus in China. METHODS: On the basis of successfully expression of human rotavirus protein G1VP7 by recombinant adenovirus vector, the authors constructed some potent recombinant adenovirus strains encoding rotavirus G2 VP7 and G3 VP7 genes which belong to the main rotavirus isolates 97S43 and 97S48. RESULTS: Replication defective recombinant adenoviruses expressing human rotavirus serotype G2 and G3 VP7 genes, named as rvAdG2VP7 and rvAdG3VP7 were successfully constructed. VP7 genes integrated into the viral genome were identified by PCR and Southern blot assay, and specific transcription were detected by RT-PCR in the 293 cells infected with recombinant adenoviruses. Expression of rotavirus VP7 proteins was demonstrated by Western blot assay. CONCLUSION: The established recombinant adenoviruses expressing G2 and G3 serotype VP7s laid a significant basis for further animal experiments in the development of multivalent rotavirus vaccines against rotavirus infection. PMID- 15340518 TI - [Antigenic and genetic characterizations of group A influenza viruses H3N2 circulated in men in China during 2000-2002]. AB - OBJECTIVE: To understand the antigenic and genetic characteristics of influenza A H3N2 viruses circulated in man in China from 2000 to 2002. METHODS: Embryonated chicken eggs inoculated with virus for amplification of viral yield. The harvesting egg allantoic fluids with influenza viruses were provided for testing antigen and RNA extraction. Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA amplified by PCR, and the product of PCR was purified. Afterward RNA sequence analysis was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers. Finally the phylogenetic tree was analyzed with MegAlign software. RESULTS: The H3N2 viruses isolated during 2000-2002 were different in amino acid sequences on HA1 domain protein molecule from those of A/Wuhan359/1995 H3N2 as well as those of A/Sydney/7/1997 H3N2 strains. There were four different positions of amino acid sequence on HA1 domain protein molecule among the H3N2 viruses isolated in 2000 and during 2001-2002. They located at 83, 186, 202, 222 and 225 position, respectively. Of them 83 and 186 were in antigenic determinant E and B, respectively. The others located at left wall of the receptor binding site (RBS). CONCLUSION: From the end of 2001 to the beginning of 2002, the influenza epidemic in Northern China caused by H3N2 virus was due to occurrence of antigenic and genetic changes of influenza A(H3N2) virus. PMID- 15340519 TI - [Baculovirus expression of two human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus]. AB - OBJECTIVE: To develop human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus (HAV) by baculovirus expression system. METHODS: The heavy and light chain genes of two human-derived neutralizing Fab antibodies to HAV were cloned into baculovirus expression vector Pac-kappa-Fc and Pac-L-Fc, and further expressed in insect cells as IgG antibodies. The IgG products were purified and well characterized. RESULTS: The baculovirus expressed McAb HAFc16 fully retained the specificity of binding to hepatitis A virus and the competition with mouse anti-hepatitis A virus McAb using ELISA. The viral neutralization assay in vitro demonstrated the retention of antibody function after expression of the human antibody in insect cells. The other expressed antibody HAFc78 also has the neutralizing activity but it is directed against different epitopes of HAV when compared with HAFc16. CONCLUSION: The recombinant baculovirus/insect cells expressed human neutralizing IgG antibodies to hepatitis A virus retained all biological functions specific for hepatitis A virus. The results provided the possibility of using these antibodies to rapidly protect high risk or early exposure populations from hepatitis A virus infection. PMID- 15340520 TI - [A study on detection method of lamivudine related mutations in hepatitis B virus polymerase gene]. AB - OBJECTIVE: To establish a simple and accurate method for rapid detection of lamivudine related mutations in hepatitis B virus (HBV) polymerase gene. METHODS: HBV polymerase gene fragments of covering B and C active region were amplified by nested polymerase chain reaction (nPCR) or nested mismatched PCR. The PCR products were digested with Nde I or Nia III and subjected to electrophoresis on agarose gel, respectively. The patterns of restriction fragment length polymorphism (RFLP) were distinguished. Using this method, thirty patients with chronic hepatitis B and treated with lamivudine for at least one year were analysed for the lamivudine related mutations in polymerase gene. Sixteen cases without lamivudine therapy were used as controls. Some of the patients were also analysed by clone sequencing. RESULTS: The nested mismatched PCR-RFLP method was simple, accurate and rapid. The whole experiments could be finished in eleven hours. The least titers of HBV DNA which could be detected was 10.3 copies/ml. The wild or mutant strains judged by RFLP were identified by clone sequencing. Mutation in the tyrosine methionine aspartic aspartic acid (YMDD) motif of HBV polymerase gene was found in eight patients and mutations of YMDD motif associated with L526M were found in another three patients. However, there were no such mutations in the control cases. CONCLUSION: The nested PCR-RFLP is considered as a simple and accurate method for rapid detection of lamivudine related mutations in HBV polymerase gene. It is suitable for larger number of sample detection. PMID- 15340521 TI - [Development of hygromycin-resistant packaging cell line for hepatitis B virus derived vectors]. AB - OBJECTIVE: To cooperate with the study of HBV vector, hygromycin-resistant packaging cell line was developed that allows encapsidation of plasmids into HBV particles. METHODS: Free of packaging signal, HBV genome was inserted into plasmid pMEP4, which expresses the HBV structural proteins including core, pol and preS/S proteins. HepG2 cell lines were employed to transfect with the construct. Hygromycin selection was done at a concentration of 150 micrograms/ml in the culture medium. The hygromycin-resistant clones with the best expressions of HBsAg and HBcAg were theoretically considered as packaging cell line and propagated under the same conditions. It was infected with recombinant retrovirus vector and hen selected with G418 and hygromycin in the culture medium. The existence of recombinant HBV virion in the culture medium was examined by PCR. RESULTS: Hygromycin-resistant HBV packaging cell line was generated, which harbored an HBV mutant whose packaging signal had been deleted. Expressions of HBsAg and HBcAg were detectable. Infected with recombinant retrovirus pRV-CP, the hygromycin-resistant packaging cell line was found to secrete mutant HBV particles and no wild-type HBV was detectable in the culture medium. CONCLUSION: After the packaging signal was deleted and transfected into HepG2 cell lines, the partial HBV genome lost its ability to form wild-type HBV, but conserves cis action providing structural proteins for the packaging of the replication defective HBV. PMID- 15340522 TI - [Clinical outcomes of 283 patients of transfusion-related hepatitis C in the northern areas of China]. AB - OBJECTIVE: To investigate the natural history and clinical outcomes in a cohort of transfusion-related hepatitis C in northern areas of China. Methods The authors studied 283 patients (137 males, 146 females, mean age 45.79+/-9.92 age) who became infected with HCV while donating plasma 12.25 years ago. These cases were subjected to ultrasonography and liver biochemical tests and serologic anti HCV assays. Statistical analyses were performed using the SPSS software. RESULTS: Ultrasonographic findings suggestive of liver cirrhosis were 8.3% of cases. No decompensated cirrhosis or HCC was detected. The value of ALT was higher in severe chronic hepatitis (mean 62.07+/-50.87 IU/L) and cirrhosis (mean 115.50+/ 108.41 IU/L) patients than in the other groups (mean 32.30+/-29.10 IU/L). The abnormal rate of ALT was 53.3% in severe group, 100% in cirrhosis group. CONCLUSION: The natural history of transfusion-related hepatitis C in the areas seemed to be relatively mild when compared with previous data. Our cases showed relatively low rate of positive findings in ultrasonography. None of this cohort had the decompensated cirrhosis or HCC. The factor of sex but not the age at time of infection was found being related to the outcomes. In the absence of liver biopsy, ultrasonography was a suitable and sensitive method for the diagnosis of the progressive hepatitis and cirrhosis. PMID- 15340523 TI - [Distribution and efficiency of recombinant adenovirus mediated hVEGF165 gene transfer in bone marrow transplanted mice]. AB - OBJECTIVE: To investigate the rapid construction of enhanced green fluorescent protein (EGFP) labeled recombinant adenovirus containing hVEGF165 and its distribution and efficiency in bone marrow transplanted mice. METHODS: The recombinant adenovirus Ad-EGFP/hVEGF165 was rapidly constructed by using AdEasy system based on the homologous recombination in bacteria, and its property was studied in vitro. Then 3x10(8) PFU adenovirus was injected into BALB/c mice via the tail vein accepted syngeneic bone marrow transplantation. The in vivo distribution of adenovirus and plasma levels of VEGF were measured at different phases. RESULTS: The adenovirus Ad-EGFP/hVEGF165 was quickly constructed by homologous recombination in bacteria using AdEasy system. The purified particles were homogenous hexagon with titers between 10(10) PFU/ml and 10(11) PFU/ml. The Hela cells infected with Ad-EGFP/hVEGF165 did not show cytopathic effects after several times passages. Under the fluorescent microscope, EGFP was revealed in the heart, lung, liver, spleen, kidney and intestine of mice at different phases. RT-PCR and immunohistochemistry showed hVEGF165 expressed significantly. No obvious damages were observed in different organs by HE staining. The plasma level of hVEGF165 was up to 866.67+/-97.13 pg/ml. CONCLUSION: These results suggested that the construction of adenovirus vector by homologous recombination in bacteria was an efficient and time-saving method, and high titer adenovirus could successfully mediate the safe and stable expression of hVEGF165 in post bone marrow transplanted mice. All these would make further gene therapy in bone marrow transplantation possible. PMID- 15340524 TI - [Replication and encapsidation of HBV mutants with the truncated C gene]. AB - OBJECTIVE: To evaluate the replication and encapsidation of HBV mutants with the truncated C gene. METHODS: The HBV mutants with the truncated C gene were constructed by molecular cloning and PCR-based deletion in vitro. The replication and encapsidation of HBV mutants were investigated by Southern blotting, PCR and real-time fluorescence PCR respectively after transfecting the HBV mutants plasmid into HepG2 cells by using liposome. RESULTS: The C-truncated HBV mutant vectors were constructed successfully and confirmed exactly by clone sequencing and enzymes digestion. The C-truncated HBV mutants were replication defective, however, all types of HBV DNA could be detected positive in the cytoplasm and supernatant after co-transfecting the C-truncated HBV mutants plasmid and the helper constructs into HepG2 cells. The C-truncated HBV mutants were proved to produce 3-40 folds more progeny DNA than that of the wild-type HBV by DNA quantitative assay. CONCLUSION: The C-truncated HBV mutants are replication deficient and could not replicate and encapsulate in the hepatocytes when transfected solely, however, the progeny HBV-variant viruses are encapsidated more effectively to secrete into supernatant when co-transfected with the helper construct which lacks part of 5 prime-proximal HBV RNA packaging signal Epsilon. PMID- 15340525 TI - [The construction of attenuated Tiantan recombinant vaccinia virus vector with IFN-gamma receptor gene deletion]. AB - OBJECTIVE: B8R gene encodes a secreted protein with homology to IFN-gamma receptor, which neutralizes the antiviral and immunological regulation activities of IFN-gamma. To improve the safety of vaccinia virus vector, an attenuated recombinant vaccinia virus with the B8R gene deletion from Tiantan vaccine strain (VTT) was constructed. METHODS: The transfer vectors were generated by joining B8R left flank, B8R right flank, vv promoter, LacZ, multicloning site and pBRSK fragments. The recombinant viruses VTTdeltaB8RLacZ (VTT with B8R deletion and LacZ insertion) were constructed by homologous recombination. RESULTS: The B8R deletion mutants were confirmed by dot blot with B8R gene probe and PCR amplification. The replication ability of VTTdeltaB8RLacZ strain in vitro was similar to that of the VTT. The skin lesions formed by VTTdeltaB8RLacZ (10(6) pfu) were significantly smaller and healed faster than those formed by VTT when injected intradermally to the rabbits,and no visible ulceration occurred. Meanwhile LacZ in VTKgpedeltaB8RLacZ was expressed stably. CONCLUSIONS: The attenuated vector with B8R gene deletion improves the safety of recombinant vaccinia virus vaccine B8R locus may be used as a new site for insertion of foreign genes in vaccinia virus vector. PMID- 15340526 TI - [Establishment of stable expression cell lines for HBsAg variants and analysis of antigenicity]. AB - OBJECTIVE: To study the mechanism of hepatitis B virus infected patients who is negative for HbsAg. METHODS: DNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified. Four of the six variants were combined point mutants and two were insertion variants. These S genes were subcloned into eukaryotic expression vector EBO-plpp, and the recombinant eukaryotic expression plasmids were transfected into COS7 cells. Cell lines expressing mutant type HBsAg were obtained. The supernatants were detected by ELISA and RIA. RESULTS: Only the two amino acid-insertion variants could be detected and the others failed to react with polyclonal and monoclonal antibodies against HbsAg. CONCLUSION: The results indicated that the point mutations and insertions may result in a conformational change of the S gene, which affect HBsAg antigenicity, suggesting a possible relationship between the variants and the negative conversion of HBsAg of the patients. PMID- 15340527 TI - [Studies on IgG antibodies of SARS patients]. AB - OBJECTIVE: To investigate the significance of detecting specific serum IgG antibodies in clinical diagnosis of SARS as well as affecting factors. METHODS: Enzyme-linked immunoassay kit for SARS coronavirus antibodies developed by HuaDa Biological Company was applied to detect specific serum IgG from SARS patients and the production of SARS specific antibodies among patients of different age groups, sex and with or without steroid treatment were statistically compared. RESULTS: Out of 121 patients studied, 71.1% were SARS specific IgG positive. Patients younger than 15 years, between 15 to 59 years, older than 59 years had positive rates of 60.0%, 70.2%, and 85.7%, respectively with no statistically significance (P=0.766); patients with or without steroid treatment showed positive rates of 70.6% and 72.4%, respectively (P=0.84); patients exhibiting either severe or light syndromes showed positive rates of 78.1% and 67.4%, respectively (P=0.493); both male and female patients showed the same positive rate of 71.1%. CONCLUSION: The sensitivity of the SARS specific IgG kit utilized needs to be further improved. The production of SARS IgG is not notably correlated with sex, age, seriousness of symptoms, and steroid treatment. PMID- 15340528 TI - [p16INK4a expression mediated by recombinant adenovirus can induce senescence of A549 cells]. AB - OBJECTIVE: To construct E1-deletion and replication-defective human type 5 recombinant adenovirus vector and to study the effect of p16INK4a on proliferation and aging of A549 cells. METHODS: p16INK4a cDNA was cloned into pAdCMV to construct recombinant pAdCMV p16INK4a, which was co-transfected into 293 cell together with pJM17. The recombinant p16INK4a adenovirus (Ad-p16INK4a) was generated by homologous recombination and identified with duplex PCR. Lung cancer cell A549, which has a homozygous deletion of p16INK4a gene, was infected with the prepared Ad-p16INK4a virus. X-gal staining and TRAP-ELISA were used for detecting senescence-associated beta-galactosidase and telomerase activities in A549 cells. RESULTS: Immunohistochemical staining and Western blot indicated that p16INK4a gene was transferred into A549 cell with more than 95% efficiency by recombinant adenovirus and p16INK4a protein was expressed at a high level- p16INK4a could markedly inhibit growth of A549 cells, induced expression of senescence-associated beta-galactosidase and suppressed telomerase activity in A549 cells. CONCLUSION: Recombinant adenovirus vector could efficiently mediate transfer and expression of foreign genes in human cell and could be used for gene immunization and gene therapy; p16INK4a could inhibit A549 cell growth and induce its replicative senescence. PMID- 15340529 TI - [CD21-independent infection of a human signet ring cell gastric carcinoma cell line by Epstein-Barr virus]. AB - OBJECTIVE: To understand Epstein-Barr virus (EBV) infection of gastric carcinoma cells. METHODS: The authors tested the infection of a signet ring cell line HSC 39 derived from human gastric carcinoma with Akata and P3HR-1 strains of EBV. Akata and P3HR-1 infected of EBV cell clones were isolated by a limiting dilution method. RESULTS: EBV-encoded small RNAs (EBERs) were expressed in the infected cells with each EBV strain by in situ hybridization. The EBV infected parental cells and most clones expressed EBNA1, but not EBNA2, latent membrane protein (LMP) 1 and LMP2A. Both EBV strains infected parental cells and clones presented type I latency. The uninfected HSC-39 cells were negative for CD21 expression; however, the Akata but not P3HR-1-infected clones were positive for CD21 expression at mRNA level. CONCLUSION: These results demonstrated that EBV infecting HSC-39 by CD21-independent pathway. This study also defined a signet ring cell line as a new target for EBV. PMID- 15340530 TI - [Antisense oligonucleotide inhibition of coxsackievirus B3 gene expression in HeLa cells and dose-response experiments]. AB - OBJECTIVE: In this study, the authors investigated inhibition of coxsackievirus B (CVB) gene expression using antisense oligonucleotides complementary to the 5' NCR, translation initiation codon and structural protein coding sequences and also observed the dose-response of the sequence specific inhibition of CVB plaque formation by antisense oligonucleotides. METHODS: Antiviral activities of these oligonucleotides were evaluated by using plaque reduction assay, yield reduction assay, cytopathic effect (CPE) and Western blot analysis. The cells were treated with random oligonucleotides as a specificity control. RESULTS: At a screening concentration of 5 micromole, 6 of the phosphorothioate oligonucleotide demonstrated some reduction of virus replication relative to untreated cells. 70% 90% inhibition of virus at 0.1 MOI (multiplicity of infection), 50% inhibition of virus infection at 10 MOI. The levels of the VP1 were reduced in CVB-infected cells treated with Scb561 and Scb733. VP1 was significantly reduced after treatment with 0.625 micromole Scb561 and almost undetectable in cells treated with 2.5 micromole Scb561. Dose response experiments implied that sequence specific oligonucleotide doses were related to effect on inhibition of CVB3 infection. When oligonucleotide doses were increased from 1.25 to 5 micromole, 75% to 90% inhibition were observed with Scb561 and 65% to 80% inhibition with Scb733, whereas random control failed to inhibit CVB replication (8% inhibition for each). CONCLUSION The present studies showed that antisense oligonucleotides against internal ribosome entry site (IRES) and translation initiation codon were capable of specifically inhibiting the synthesis of viral protein and subsequent productive CVB replication.The selective inhibition using antisense oligonucleotide might lead to development of an effective antiviral agent for future clinical evaluation. PMID- 15340531 TI - [Expression of human cytomegalovirus immediate early gene in the intracranial artery walls of atherosclerosis]. AB - OBJECTIVE: Human cytomegalovirus (HCMV), especially the immediate early (IE) gene of the virus, has been implicated in the pathogenesis of atherosclerosis. The aim of this study was to confirm the presence of HCMV IE gene DNA in intracranial artery walls and the association of the virus with the development of atherosclerosis. METHODS: HCMV IE gene was tested in formaldehyde-fixed intracranial arteries from 35 cases with cerebral atherosclerosis and 20 negative controls. In situ hybridization as well as polymerase chain reaction (PCR) was used to detect the presence of DNA in sections of paraffin-embedded tissue samples. Probes and primers were derived from major immediate early (MIE) genomic regions of cytomegalovirus strain AD169. RESULTS: The DNA of HCMV was found in 40.0% and 10.0% of arterial walls with atherosclerosis and negative control group by in situ hybridization, respectively, in 60.0% and 30.0% by PCR, respectively. Significant deference was found between them (P=0.018, P=0.032). There was also significant difference between grade III-IV and grade I-II atherosclerosis by both methods (P=0.027, P=0.009). CONCLUSION: The results suggested that HCMV IE DNA exists in the atherosclerotic arterial walls, therefore, there might be an association between the IE gene in intracranial artery walls and the atherosclerosis. The arterial wall with the smooth muscle cells, might be the potential site of the virus persistence. HCMV may play a role in the pathogenesis of the atherosclerosis. PMID- 15340532 TI - [Clinical trial of sequential antiviral therapy for patients with chronic hepatitis B in China]. AB - OBJECTIVE: To establish a sequential antiviral regime and evaluate its efficacy in patients with chronic hepatitis B using a controlled trial. METHODS: Seventy four patients with chronic hepatitis B were divided into 3 groups: 30 cases were enrolled in the sequential antiviral group in which patients received eight-week treatment with thymosin alpha1 (1.6 mg/time, subcutaneous injection, 2 times/week), six-month treatment with interferon (500 MU/ times, muscle inject, every other day) begun in the fifth week of the therapeutic course, and lamivudine treatment (100 mg/days) begun 2 months later after HBeAg seroconversion or just after the withdrawal of interferon to more than eighteen months. Fourteen cases were enrolled in combination group in which patients received six-month treatment with interferon and thymosin alpha1 simultaneously in the same manner as in sequential antiviral group. Thirty cases were enrolled in lamivudine group in which patients received more than eighteen-month treatment with lamivudine. RESULTS: The temporary rates of HBeAg seroconversion and normalization of alanine aminotransferase (effective rate) in sequential antiviral group, combination group and lamivudine group were 76.7%, 78.6% and 13.3%, respectively. The effective rates of sequential group and combination group were very similar, and significantly higher than that of lamivudine group (P less than 0.01). Long-term efficacy rates were 76.7%, 57.1% and 16.7% among the three groups, respectively. The long-term effective rate of sequential group was relatively higher. The rate of liver damage sensitive period in sequential antiviral group and combination group was 47.7%. The time of onset was from 2 to 8 weeks after the treatment begun, earlier than that from 6 to 8 weeks after the beginning of interferon alone in the literature. CONCLUSION: Sequential antiviral therapy had much higher rates of long-term HBeAg seroconversion, undetectable HBV DNA and normalization of alanine aminotransferase with good cost-effectiveness. Its mechanism to promote the antiviral effect might be dependent on the immunoregulatory action of thymosin alpha1 in the earlier period and the specific inhibition of HBV DNA replication by lamivudine in the later period of the therapeutic course. PMID- 15340533 TI - [Clinical study on patients at early stage of severe acute respiratory syndrome]. AB - OBJECTIVE: To elucidate the clinical features of patients at early stage of severe acute respiratory syndrome (SARS). METHODS: Fifty-three cases of early SARS were studied retrospectively. The data reviewed included those of epidemiology, clinical manifestations, laboratory investigation and roentgenology. RESULTS: The patients consisted of 24 men and 29 women, aged 10.85 years (mean 38+/-16.7 years), including 9 infected health-care professionals (17.0%). The mean incubation period was 7.3+/-7.0 days (3.14 days). The onset symptoms were characterized by fever (100%), cough (49.1%), maylgia (24.5%), shortness of breath (20.8%), malaise (17.0%),and diarrhea (5.7%). Routine blood test during the first to the fifth day of the disease revealed WBCs less than 4.0x10(9) /L in 33 patients (62.3%), 4.0-10.0x10(9)/L in 18 patients (34.0%), lymphopenia in 36 patients (67.9%), and PLT less than 100.0x10(9) in 7 patients (13.2%). The main abnormal X-ray finding was single (75.4%) or bilateral (15.1%) localized patchy clouding opacity. The decrease of arterial partial pressure of oxygen occurred in 26 patients (49.1%). The damage of several organs was common, including the elevated ALT or AST in 20 patients (37.7%), elevated BUN or SCR in 6 patients (11.3%) and elevated LDH or CK or HBDH in 23 patients (43.4%). CONCLUSION: The clinical manifestations of SARS at the early stage were complex. The close monitoring of the blood cell counts, the blood gas analysis and chest radiography might be crucial to the early diagnosis. PMID- 15340534 TI - [Clinical investigation on the treatment of HCMV hepatitis in children]. AB - OBJECTIVE: To investigate the effective therapeutic method of human cytomegalovirus (HCMV) hepatitis in children. METHODS: Twenty-five children with HCMV hepatitis were randomly assigned to a treated group (n=13) or a control group (n=12). Both groups were treated with prednisone, glucurone, luminal and Xiaoyanlidanpian. But the treated group was given ganciclovir (GCV) + intravenous immunoglobulin (IVIG) in addition. Each infant of the two groups was checked for blood routine, liver function and HCMV copy numbers on admission and before discharge. They were seen at the third, sixth and ninth month after discharge. On each visit blood specimens were collected for HCMV copy numbers (fluorescence quantitative PCR, FQ-PCR). RESULTS: The viral load of the treated group decreased significantly. A significant difference in viral copy numbers was found between the two groups on admission, discharge, and third, sixth and ninth month after discharge (P less than 0.001). The number of HCMV DNA copy fell to 10(3) copies/ml on discharge while that of the control group fell to the same level after the third month. The differences between the two groups in the length of hospitalization, time of initial jaundice disappearance and complete disappearance were statistically significant (P less than 0.05). The need for transfusion in the treated group was significantly less than that in the control group (chi-square=4.012, P less than 0.05). CONCLUSION: Combination of GCV with a high dosage of IVIG to treat HCMV active infection could decrease viral load remarkably; The duration of disease, severity of symptoms, degree of anemia and the need for blood transfusion were reduced. No adverse effects related to the combination of GCV with IVIG therapy were observed. PMID- 15340535 TI - [IFN or oxymatrine in combination with lamivudine in patients with lamivudine resistant chronic hepatitis B]. AB - OBJECTIVE: To observe the therapeutic efficacy of IFN or oxymatrine in combination with lamivudine in patients with lamivudine-resistant chronic hepatitis B. METHODS: Forty patients ongoing treatment with lamivudine were randomized to three groups: group A, 14 patients with addition of IFN alpha-2b 3MU to ongoing lamivudine, daily, one month, followed by the same dose given every other day, five months; group B, 15 patients with addition of injectable oxymatrine 60 mg daily, three months, followed by oral oxymatrine every day, three months, and group C, 11 patients ongoing treatment with lamivudine alone. The HBV DNA level in serum, HBeAg seroconversion, and ALT level were detected at the end of the treatment. RESULTS: After 6 months of treatment, HBV DNA became negative in 35.73% patients treated with combination with IFN, and in 13.3% patients treated with combination with oxymatrine. ALT level was normal in 85.71% or 86.66% of patients, respectively. In none of the patients under ongoing treatment with lamivudine alone HBV DNA or HBeAg became negative, and ALT level was normal in 36.36% of patients. CONCLUSION: These data indicated that IFN or oxymatrine in combination with ongoing lamivudine therapy provided effective antiviral therapy in patients with lamivudine-resistant HBV. The addition of IFN or oxymatrine to ongoing lamivudine therapy in lamivudine-resistant patients led to significant inhibition of viral replication and improvement in liver function after 6 months of therapy. PMID- 15340536 TI - [Development of a subtype screening assay for human immunodeficiency virus type 1 by nested multiplex PCR]. AB - OBJECTIVE: The current available assays for HIV subtyping, such as sequence-based phylogenetic analysis or heteroduplex mobility assay (HMA), are labor-intensive and time-consuming. The authors have just developed a simple and rapid subtype screening assay for subtypes B, C, and CRF01-AE using a single nested multiplex PCR. METHODS: Proviral DNA from HIV-positive samples was extracted and subjected to first round PCR with universal primers for gag region that can detect HIV-1 M group isolates. In the second round PCR, three pairs of subtype-specific primers, respectively detecting subtype B, C and CRF01-AE, were added into one tube. The PCR products of different subtypes could be distinguished in agarose-gel electrophoresis. Another pair of primers exclusively detecting the prevalent recombinant B/C strains CRF07-BC and CRF08-BC were designed and used. Additionally, all of these samples were sequenced and analyzed phylogenetically. RESULTS: Phylogenetic analysis showed that out of 119 samples, there were 43 subtype B samples (Euro-American B 11, Thailand B 32), 54 subtype C, 17 CRF01-AE, 3 subtype A and 2 subtype D samples. The subtype B, C, and CRF01-AE specific primer sets detected 35 (81.4%), 46 (85.2%), and 13(76.5%) samples with accuracy and specificity. Non-specific bands occasionally appeared but did not interfere with interpretation of the results. The primer pairs for CRF07-BC and CRF08-BC amplified target sequences were confirmed by sequencing and phylogenetic analysis. The specificity of all these subtype-specific primers was found to be 100%. CONCLUSION: A simple and rapid assay was developed for screening subtypes B, C, CRF01-AE, CRF07-BC and CRF08-BC in China. This assay may have potential application in HIV laboratories in China and worldwide. PMID- 15340537 TI - [Experimental study on enzyme dot assay for detection of hemorrhagic fever with renal syndrome antigen]. AB - OBJECTIVE: To establish a new and efficient method(IEDA) for detection of hemorrhagic fever with renal syndrome virus (HFRSV) antigen. METHODS: An immune enzyme dot assay (IEDA) with mixture of three sorts anti-HFRSV-IgG, which was obtained from rabbit vaccinated with EHFV R22, Chen and Hubei strain was employed to detect HFRSV antigen in serum and urine from epidemic hemorrhagic fever (EHF) patients, and compared with indirect immune fluorescence assay (I-IFA), 76 serum samples and 40 urine samples were detected in this study. RESULTS: The results showed that the total positive rate of HFRSV antigen detected by IEDA was 73.68% in serum and 65.00% in urine, while that detected by I-IFA was 75.00% and 70.00%, respectively. The positive rate in primary phase (within 5 days) of HFRSV antigen detected by IEDA was 94.34% in serum and 83.33% in urine, while that detected by I-IFA was 64.42% and 55.56%, respectively, there was significant difference in both serum and urine detections. Correlation study showed a high correlation in the result of IEDA and I-IFA. CONCLUSION: The results of this study suggested that the IEDA, as compared with I-IFA, was a more specific, sensitive, rapid and simple method with higher positive rate in primary phase. IEDA could be widely used for early diagnosis of HFRS in hospital at grassroots level. PMID- 15340538 TI - [Sequencing of adenovirus type 7 vaccine strain fragment and characterization of the hexon encoding gene]. AB - OBJECTIVE: To complete the full-length sequencing of the human adenovirus type 7 vaccine strain (Ad7v) for novel vector constructing. METHODS: The Ad7v DNA was digested with SalI and the 17.5-68.0 map unit (mu) fragment was cloned and sequenced. The homology of encoding sequence of Ad7v hexon to those of group A,C,D,E,F and other numbers of group B was accomplished with the software CLUSTAL.V. The three-dimensional structure of the Ad7v hexon was predicted with the RasMo12.71. RESULTS: The fragment contains 17,596 bp, part of E2 and late gene L1, L2 and L3 were encoded by this region. Polypeptide encoded by hexon gene lies in L3 region, which is composed of 934 amino acids. Multiple sequence alignment with the other nine known hexon protein sequences suggested that the variable sequences are mainly concentrated on seven regions, namely hypervariable regions (HVRs). The seven HVRs are related to type-specificity and group specificity. The three-dimensional structure of the Ad7v hexon revealed that the variable regions are located in the I1 and I2 loops of the molecule mostly on the tower of the hexon. CONCLUSION: The full-length genome sequencing of Ad7v was accomplished at last. Since the deduced amino acid sequence of Ad7v hexon was quite different from other adenoviral vectors such as Ad5 and Ad2, this virus can be potentially used for the construction of novel gene delivery vectors to counterpart the immunity to the vectors widely used at present. PMID- 15340539 TI - [Gene optimization is necessary to express HPV type 6 L1 protein in the methylotrophic yeast Pichia pastoris]. AB - OBJECTIVE: Human papillomavirus 6 (HPV 6) causes genital warts, a common sexually transmitted disease. L1-capsids protein is a highly promising vaccine candidate that has entered phase II clinical trial. But the existing methodologies for producing L1-capsids in insect cells is expensive for use in developing countries. As methylotrophic yeast,the Pichia pastoris expression system offers economy,and high expression levels. Over-expression of HPV6-L1 protein in P. pastoris was the purpose of this study. METHODS: The whole L1 gene with preferred codons for P. pastoris was rebuilt and A-T rich regions were abolished, Cloning into pPIC3.5K,electroporation of KM71, in vivo screen of multiple inserts by G418 resistance, PCR analysis of pichia integrants, BMGY/BMMY are used for induction and expression of L1 proteins. RESULTS: Three clones were found to produce L1 protein which can be identified with Western blot. Compared with L1 protein from E.coli, pichia-produced L1 has some glycosylation. Reacting strongly with MabH6B10.5 in indirect immunofluorescence assay indicated that L1 protein expressed in pichia cell holds its native conformational epitopes which is important for vaccine use. A total 125 microg pure L1 protein could be obtained from 1L cultures through ion-exchange and Ni-NTA chromatography. CONCLUSION: HPV type 6 L1 protein expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study. PMID- 15340540 TI - [Origin of hemagglutinin and neuraminidase gene of swine influenza A H1N1 viruses]. AB - OBJECTIVE: To understand the origin of hemagglutinin (HA), and neuraminidase (NA) gene of swine influenza A (H1N1) viruses isolated in pigs in mainland China in 2002 and reveal the reason of pathogenesis of them in pigs. METHODS: The target gene amplified by PCR,PCR product was linked with PGEM-T Easy Vector(Promega company, USA) at 4? degrees C, the recombined plasmid was transferred into DH 10B bacteria, positive colonies were selected and identified them with restriction enzyme. Afterwards, they were sent to Liu He Tong company in Beijing for testing nucleotide sequence. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (Version 1.03)and Editseq (Version 3.69) software. RESULTS: The HA and NA genes of three strains of swine influenza A (H1N1) viruses isolated from pigs in China were closely related to those of swine influenza A (H1N1) virus, but different from those of avian or human influenza A (H1N1) virus. The swine strain of influenza A (H1N1) virus isolated in 2002 was derived from swine influenza A (H1N1) virus circulated in pigs in China in 1991. Since the antigenic drifts of HA and NA proteins of the new isolates occurred, their activity in pigs is increasing and they can cause disease in pigs. CONCLUSION: The HA and NA genes of three strains of influenza A (H1N1) virus tested were identified to be derived from those of swine influenza A (H1N1) virus. The increased activity and pathogenesis of them in pigs were most likely due to antigenic drifts of HA and NA proteins of the new isolates. PMID- 15340541 TI - [Capacity of HIV enzyme immunoassay diagnostic kits to detect antibodies against different genotypes of HIV]. AB - OBJECTIVE: To investigate the capacity of commercial HIV enzyme immunoassay (EIA) diagnostic kits to detect antibodies against different genotypes of HIV. METHODS: HIV RNA was detected with RT-PCR from samples positive for HIV antibody. The purified PCR products were sequenced directly and the genotypes of HIV from samples were analyzed. The samples for each genotype of HIV were diluted and the diluted samples were detected with different HIV EIA diagnostic kits. RESULTS: All 20 samples positive for HIV antibody were also positive for HIV RNA; 9 of 20 isolates were genotype B, 9 of them were genotype C or CRF BC, 2 of them were CRF AE. The sensitivity of different HIV EIA diagnostic kits to detect antibodies against different genotypes of HIV was not significantly different. CONCLUSION: The capacity of commercial HIV diagnostic kits to detect antibodies against different HIV genotypes may not be significantly different. PMID- 15340542 TI - [Investigation of Arboviruses in Lancang river down-stream area in Yunnan province]. AB - OBJECTIVE: To investigate the epidemic state of arboviruses in the downstream area of Lancang river in Yunnan province. METHODS: Mosquitoes were collected from Lancang river downstream area (including Lancang county and Simao city) during summer in 1998 and stored in liquid nitrogen after classification. The mosquito pools were homogenized and centrifuged, then the supernatant was inoculated into C6/36 cells for virus isolation. New isolates were identified by neutralization test(NT), ELISA, immunofluorescence assay(IFA) and polyacrylamid gel electrophoresis(PAGE). RESULTS: Totally 22 isolates of arbovirus were obtained from 233 mosquito pools by inoculation of C6/36 cells and positive rate of the isolation was 9.4%. Ten strains were resistant to both ether and 5 prime-IDU. So they were non-enveloped double-stranded (ds) RNA virus. Twelve segmented RNAs were shown by PAGE and PAGE profiles from the ten strains were 6-6 with minor variation. The isolates can be neutralized by immunized mouse ascites fluid of BJ95-75 strains of coltivirus by NT, and reacted with monoclonal antibody against BJ95-75 by ELISA. These viruses were identified as coltivirus. Nine isolates were sensitive to ether and resistant to 5 prime-IDU. So they were non-enveloped RNA viruses. PAGE showed 10 segmented RNA, and they were identified to be orbiviruses. Three isolations were sensitive to ether. One of them can be neutralized with JEV A2 strain antibody by NT and was positive to the homologous antibody by IFA. It was identified being strain of JE virus. One strain(YN92-4) can be reacted with anti-bunyavirus group specific immune ascites fluid by both IFA and ElISA, but reacted neither with anti-alpha virus group, nor with anti flavivirus group JE virus ascites fluid. The virions are spherical and about 87 nm in diameter with surface projections by negative staining. Conclusion Twenty two isolates have been obtained from wild caught-mosquitoes of Lancang river down stream area in Yunnan province. Among them ten, nine, one and one were identified as coltivirus, orbivirus, JE virus and bunyavirus, respectively. One is under identification. This is the first report on bunyavirus isolated from mosquitoes in China. PMID- 15340543 TI - [Comparison of hepatitis B virus serotype and genotype among HBsAg positive hepatitis B patients in a northern and a southern city of China]. AB - OBJECTIVE: To understand HBV serotypes and genotypes epidemiology in a northern city and a southern city in China. METHODS: Using polymerase chain reaction (PCR) and direct sequencing of HBV DNA PCR products, the serotypes and genotypes of HBV in 530 from HBsAg positive samples. The enrolled patients were from Harbin, a northern city and Lianjiang, a southern city in China. RESULTS: Comparison of the serotypes and genotypes of HBV between Harbin and Lianjiang showed that adrq+ was the most predominant hepatitis B virus serotype in both Harbin and Lianjiang (87.2% and 73.5%,respectively), adw2 was the next (12.0% and 25.7%, respectively); genotype C was the most frequent in Harbin and Lianjiang (87.8% and 73.2%, respectively), and genotype B was the next (12.2% and 26.1%, respectively) only 1 patient was infected by genotype D, and 1 patient was found to be co-infected by genotype B and C in Lianjiang. CONCLUSION: The results suggest that the percentage of HBV serotypes and genotypes between Harbin and Lianjiang was significantly different (P less than 0.001), but the main HBV serotype and genotype of the two cities were similar. PMID- 15340544 TI - [Detection of antibody against Borna disease virus-p24 in the plasma of Chinese patients with chronic fatigue syndrome by Western-blot analysis]. AB - OBJECTIVE: To evaluate the prevalence of infection with Borna disease virus (BDV) in Chinese patients with chronic fatigue syndrome (CFS) and control subjects, and to discuss the etiological association between CFS and infection with BDV. METHODS: The CDC (1994) diagnostic criteria for CFS were used for case definition. Sixty-one patients suffered from CFS were from 11 Provinces in China. To detect the antibody against BDV-p24 on the plasma samples from all cases and 73 healthy control subjects by Western-blotting analysis. RESULTS: 7 of the sixty one cases and 0 of the controls were sero-positive for BDV-p24 antibody, there was a statistical significant difference between the two groups (11.48% vs 0%; P less than 0.010). CONCLUSION: Chinese patients with CFS showed sero-positive identifying BDV infection, by comparison, anti.BDV-p24 antibody prevalence in patients was significantly higher than in controls. An etiological association may exist between CFS and BDV infection. PMID- 15340545 TI - [Relationship between human herpesvirus 6 infection and oral squamous cell carcinoma]. AB - OBJECTIVE: To study the relationship between human herpesvirus 6 (HHV-6) and oral squamous cell carcinoma. METHODS: The serum anti-HHV-6 antibody titers from oral squamous cell carcinoma patients and control subjects were detected by indirect immunofluorescence assay. HHV-6 DNA in peripheral blood mononuclear cells from oral squamous cell carcinoma patients and control subjects was amplified by PCR with primers from sequence of HHV-6 and the specificity was confirmed by Southern blot hybridization with an internal probe oligonucleotide. An immunohistochemical staining using rabbit anti-HHV-6 antibody was used to detect HHV-6 antigen in oral tumor tissues from oral squamous cell carcinoma patients. RESULTS: Significantly higher proportion of patients with oral carcinoma (16/16) had IgG antibody to HHV-6 in sera compared with those (12/16) in control subjects, and geometric mean titer of these two groups was 1:118 and 1:64 respectively (P less than 0.05). The detectable rate of HHV-6 DNA in peripheral blood mononuclear cells for the above groups was 10/16 and 6/16 respectively (P less than 0.05). HHV-6 antigens were positive in 9 out of 12 oral tumor cases and in only 2 out of 8 pericancerous tissues the difference between these two groups was also significant (P less than 0.05). CONCLUSION: These results demonstrated the frequent presence of HHV-6 in oral squamous cell carcinoma, therefore, HHV-6 possibly play a role in the pathogenesis of oral squamous cell carcinoma. PMID- 15340546 TI - [Relationship between ultrasonic findings and pathological features in patients with chronic viral hepatitis B and C]. AB - OBJECTIVES: To explore the relation of ultrasonic findings to pathological features in cases of chronic viral hepatitis B and C. METHODS: The ultrasonic and pathological findings were analyzed in 130 patients with chronic viral hepatitis B and 106 with chronic viral hepatitis C. RESULTS: In patients with hepatitis B, the ultrasonic echo was thicker and more intensive and uneven cords were found. These findings were closely related to the pathological findings (P less than 0.001). In those with hepatitis C, the ultrasonic echo was slight and dense, which was also closely related to the pathological findings (P less than 0.001). In the patients complicated with fatty liver, the ultrasonic findings were also different (P less than 0.001). CONCLUSION: Ultrasonography is helpful for differential diagnosis of hepatitis B and hepatitis C. PMID- 15340547 TI - [Detection and identification of emergence of HBV YMDD motif mutation during lamivudine treatment by hybridization to an oligonucleotide array]. AB - OBJECTIVE: Gene chip technology was set up to quickly and accurately detect and identify the HBV P gene YMDD motif mutation during the chronic hepatitis treated with lamivudine. METHODS: DNA microarrays were prepared by spotting fluorescence labeled probes of target genes onto specially lattice glass slides with robotics. The serum samples from 30 patients with hepatitis B after 68-week treatment with lamivudine were detected double-blind by gene chips and by nucleotide sequences assay technique to identify the rate of emergence of HBV P gene YMDD motif mutation. RESULTS: Twenty-one patients were found HBV P gene YMDD motif mutation by the gene chips including 11 cases with YVDD and 10 cases with YIDD motif mutation. By direct sequencing of the PCR products, 11 cases were found to have YVDD with adenine741 changed into cytidine resulted in methionine552 changed into valine in which 6 cases with adenine669 changed into cytidine and leucine changed into methionine, 10 cases had YIDD motif mutation with guanosine743 altered thymidine methionine552 changed into isoleucine, including 3 cases with thymidine281 changed into cytidine and leucine565 altered proline. CONCLUSION: The gene chip can be used to test HBV YVDD,YIDD motif mutation compared with nucleotide sequences assay technique, the accuracy rate was 100%. PMID- 15340548 TI - [Changes of blood cells in patients with severe acute respiratory syndrome]. AB - OBJECTIVE: To investigate the changes of blood corpuscles of patients with severe acute respiratory syndrome (SARS) in Beijing. METHODS: Totally 43 patients (21 male and 22 female, 19-74 years old age range) diagnosed as of probable SARS were included in this study. Their corpuscles in the peripheral blood were tested every two days, and the results were analyzed. RESULTS: Patients with SARS were more likely to develop leukocytopenia, lymphopenia and thrombocytopenia in the early period of disease than those in control group. The situation, especially lymphopenia and thrombocytopenia, could not be reversed in patients who died. Persistent low counts of lymphocytes and platelets at presentation might be associated with adverse outcomes. CONCLUSION: Low counts of leukocytes, lymphocytes and platelets were common among patients in the early stage of SARS. Persistent Lymphopenia and thrombocytopenia may be associated with the prognosis. PMID- 15340549 TI - [Genotyping of the Chinese isolates of coltivirus]. AB - OBJECTIVE: To classify the Chinese isolates of Coltiviruses. METHODS: Three sets of primers were selected among them two were specific to the 9th and 12th segments of subgroup B2, and one was for the 12th segment of subgroup B1-All the Chinese isolates of Coltivirus selected in the experiment were classified according to the lengths of different amplicons of the reverse transcriptase polymerase Chain reaction (RT-PCR). The homogenicity of the nucleic acids of the isolates BJ95-75 and YN-6 was also compared with other Coltivirus strains belonging to subgroup B2. RESULTS: With the primers 12-854-S/12-B2-R, which were specific to the 12th segment of Coltivirus subgroup B2-850 bp amplicons were obtained from Beijing isolate BJ95-75 and all the Yunnan isolates such as YN-6, 67-1, -68-1, -69, -70-1, -70-2, -90, -92-2, -93 of Coltivirus 492 bp DNA fragments were also amplified from all of them with the segment 9th specific primers 9-JKT-S/9-JKT-R. However no positive results were obtained from Northeast isolates NE97-12, NE97-31 and control viruses YN-99(Orbivirus),YN-151-1(JEV) with the same two sets of primers. With 12-B1-S/12-B1R primers specific to the 12th segment of subgroup B1, no amplicons of right length were obtained from any of the Chinese isolates of Coltivirus and the control viruses. When compared the nucleic acid sequences of BJ95-75 and YN-6 with other Coltivirus strains such as Bannavirus, JKT6423, JKT6969, JKT7043, the amplicons from segment 12th of these two strains had more than 89.4% homology with the other strains, especially to the earlier Chinese isolate Bannavirus, the homolog was more then 98.9%. Nearly 96.5% and 99.2% of the nucleic acids of the amplicons from segment 9th of the two strains were being homologous to Bannavirus and about 84.0% to JKT6423, which had been classified into type B2a. But the maximal homogenicity was about 53% when compared with the other two coltivirus strains. JKT6969 and JKT7043 which had been classified into type B2b. CONCLUSION: Genotyping the recent Chinese isolates of coltivirus for the first time in our country. Most of the Chinese isolates belong to subgroup B2, more exactly type B2a. The Northeast isolates NE97-12 and NE97-31 were not correctly grouped with the available primers. PMID- 15340550 TI - [Role of cytomegalovirus infection in the pathogenesis of type 2 diabetes mellitus]. AB - OBJECTIVE: To further study the role of human cytomegalovirus (HCMV) infection in the pathogenesis of the type 2 diabetes mellitus. METHODS: HCMV DNA levels in sera from 29 type 2 diabetic patients and 23 controls were measured by quantitative polymerase chain reaction (PCR), and comparative analyses was made between HCMV DNA and T cell subsets, blood glucose (BG), insulin (Ins) and C peptide (C-P). RESULTS: The levels of HCMV DNA were (1.81+/-1.67) x 10(8) copies/ml for type 2 diabetic patients, a level significantly higher than that (5.50+/-4.30) x 10(7) copies/ml of controls. The percentage of CD8 for type 2 diabetic patients with positive HCMV DNA was 29.53%+/-2.00%, being much higher than that for controls (27.13%+/-4.12%), while the ratio of CD4/CD8 1.24+/-0.05 was significantly lower than that 1.41+/-0.10 of controls. Fasting C-P of type 2 diabetic patients with positive HCMV DNA was far lower than that of those with negative HCMV DNA, but the differences of BG and Ins between the two groups were not significant. CONCLUSION: Active HCMV infection of type 2 diabetic patients may suppress cellular immunity and its influence on the pathogenesis of the type 2 diabetes mellitus should be further studied. PMID- 15340551 TI - [Nucleotides sequence analysis of EV71 Chinese strain SHZH98 3C segment]. AB - OBJECTIVE: To study the EV71 Chinese strain SHZH98 and analyze its genetic evolution using 3c gene as index. METHODS: The 3C gene cDNA of EV71 Chinese strain SHZH98 was amplified by PCR, the PCR product was sequenced. RESULTS: The EV71 Chinese mainland strain SHZH98 3C segment was 549 bps in length. Comparison of nucleotide sequences from other enteroviruses which have been published, revealed a higher homology to strain MS, 78.7% at nucleotide level and 93.45% at deduced amino acid level. The homology to strain BrCr was 76.7% at nucleotide level and 89.1% at deduced amino acid level. Taiwan strains POLY,NCKU,TW2086,TW2272 shared a lower homology with Chinese mainland strain SHZH98, 74.0%, 73.8%, 71.9%, 69.8% at nucleotide level and 90.7%, 90.2%, 84.2%, 82.5% at deduced amino acid level. The genetic progress analysis revealed that EV71 Chinese mainland strain SHZH98 3C segment shares more homology with European and American strains than Taiwan strains. CONCLUSION: The non-structural protein of EV71 Chinese strains may have different evolutionary process from Taiwan strains. PMID- 15340552 TI - [Construction of a tRNAVal promoter plasmid expressing shRNAs in mammalian cells which mediates RNA interference]. AB - OBJECTIVE: Constructing a plasmid containing tRNAVal promoter to express shRNA which mediates RNA interference. METHODS: A tRNAVal gene was amplified from human genomic DNA by PCR and replaced the last several bases of 3' end by a linker. The tRNAVal promoter after artificial mutation followed a shRNA sequence to luciferase was cloned into pUC18, Puc-tRNAVal, lucRi Cotransfected with pMAMneoLuc into BHK-21 cell to detect the effect of luciferase expression. RESULTS: pUC-tRNAVallucRi suppressed the luciferase expression from pMAMneoLuc by 97.9%-9.5%. CONCLUSION: The results showed that the tRNAVal shRNA plasmid could efficiently suppress luciferase expression in BHK-21. PMID- 15340553 TI - [Serological characteristics of a hepatitis E outbreak]. AB - OBJECTIVE: To look into the serological characteristics of a hepatitis E outbreak. METHODS: Sera from the first five patients with acute icteric hepatitis who developed the disease successively within ten days and the 1,675 employees routinely having their lunch in a dining hall of a department (outbreak population) were examined for anti.HEV IgM and IgG at 26th days after the outbreak, and the 883 employees of a neighboring department not having their lunch in the hall were selected as control (control population). RESULTS: The five patients were all positive for anti-HEV IgM and IgG. The positive rates of anti-HEV IgM and IgG in outbreak population were 8.7% and 38.4% respectively, both significantly higher than those in control population which were only 0.1% and 28.6%. The numbers with abnormal ALT in the 145 individuals with anti-HEV IgM(+) of outbreak population were significantly higher than those in the IgM(-) individuals of the same group as well as in control, while the abnormal ALT ratio in the IgM(-) individuals of the outbreak was not higher than that in control. The results from the four patients' serial sera showed that the anti-HEV IgM titers declined gradually and were undetectable at about 4th month after infection, and the IgG titers increased to peak in about 2-3 months after infection, then declined very slowly. The mean IgG titer of the anti-HEV IgM(+) individuals was significantly higher than that of the IgM(-) but IgG(+) individuals in outbreak population, and the latter was significantly higher than the IgG(+) individuals in control, which suggested that the post-infection individuals' immunities to HEV were boosted during the outbreak. There was no difference between sex or age groups for the anti-HEV IgM(+) ratio, but the abnormal ALT was much more frequent in the anti-HEV IgM(+) male than in the female, and no difference was observed between age groups. CONCLUSION: The pathogen of the outbreak of acute icteric hepatitis was hepatitis E virus and associated with food intake. Anti-HEV IgM and IgG were used not only for diagnosis of hepatitis E but also for surveilance in mass population. The attack risk was not associated with age or sex, but the abnormal ALT was much more frequent fresh infectors in male. PMID- 15340554 TI - [Antiviral effect of human CTLs activated by HBsAg stimulated dendritic cells in vitro]. AB - OBJECTIVE: To investigate the ability of human dendritic cells (DCs) inducing specific T lymphocyte response and inhibit the expression of HBeAg and HBsAg in 2.2.15 cell culture supernatant. METHODS: DCs were prepared from peripheral blood mononuclear cells induced with granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin 4. DCs was impulsed with pure HBsAg before DCs maturation and cocultured with self-blood T lymphocyte, while DCs without pure HBsAg stimulated group, T lymphocyte group and only T lymphocyte group were prepared as control group. The culture supernatant of 2.2.15 cell with stimulated T lymphocytes was collected on day 1, day 3, day 5 and day 7, respectively. The expressed levels of HBeAg and HBsAg were detected by ELISA method. RESULTS: DCs after antigen stimulation had a strong ability to present antigen and induce immune activation, DCs after loading with antigen in normal control and chronic hepatitis patients group had stronger stimulative ability for T lymphocytes proliferation than that of DCs without loading with antigen and only T lymphocyte group(P less than 0.01). The stimulating ability of DCs had a positive correlation to the dosage of loaded antigen; CTLs produced as a result of DCs stimulation had a specific inhibitive effect on the expression of HBeAg in 2.2.15 cell supernatant,but not on the expression of HbsAg. CONCLUSION: Human dendritic cells stimulated with HBsAg in vitro can efficiently present antigen and stimulate the production of specific CTLs to HBV, which can efficiently inhibit the expression of HBeAg in 2.2.15 cell supernatant- DC vaccine may become an antiviral therapy strategy for chronic hepatitis B virus infected patients in future. PMID- 15340555 TI - [Preliminary studies on pathogenic factors of human cytomegalovirus infection]. AB - OBJECTIVE: To investigate the pathogenic factors of human cytomegalovirus (HCMV) infections. METHODS: Totally 36 serum samples were obtained from early pregnant woman and examined with ELISA for anti-HCMV antibody IgG and IgM. After artificial abortion,chorionic villus and decidua were also examined with polymerase chain reaction (PCR) for HCMV-DNA. When the results of PCR were positive, pathological changes of these chorionic villus and decidua were analyzed. RESULTS: The results showed that only 10 samples were PCR positive while IgG and/or IgM antibody to HCMV was positive. After infection with HCMV, different changes occurred in chorionic villus and decidual trophoblastic cells placental villus were hyperplasic and decidua cells degenerated and necrotized followed by lymphocytes infiltration. CONCLUSION: These pathological changes may be one of pathogenic factors of HCMV. PMID- 15340556 TI - [Relationship between glucose metabolic disorders and expression of insulin receptor in posthepatitic cirrhosis hepatocyte and HBV DNA in pancreatic cells]. AB - OBJECTIVE: To investigate relationship between glucose metabolic disorders and expression of insulin receptor (IR) and tyrosine protein kinase (TPK) in posthepatitic cirrhosis hepatocyte and HBV DNA expression in pancreatic cells. METHODS: To detect HBV DNA in paraffin-embedded pancreatic and hepatic tissues from 12 posthepatitic cirrhosis patients with positive serum HBV markers by using in situ hybridization (ISH) with a digoxigenin labelled probe. The amount of IR and TPK have been evaluated by immunohistochemical quantitative analysis using image analyzer in hepatocyte of 12 patients positive for HBV markers with posthepatitic cirrhosis in serum. Immunofluorescent histochemical double staining technique was used. HBsAg and IR were observed under confocal laser scanning microscope. RESULTS: Eleven of 12 cirrhosis patients? hepatocytes were HBV DNA positive, including 7 patients (7/7) with impaired glucose tolerance (IGT) and 4 patients (4/5) with normal glucose tolerance (NGT). Eight of 12 pancreatic cells were HBV DNA positive, including 7 patients (7/7) with IGT, but only one patient (1/5) with NGT-HBV DNA was found positive in pancreatic cells in significantly more subjects in IGT group than in NGT group (P less than 0.01).IR and TPK amount in hepatocyte of IGT was significantly less than that of NGT patients with posthepatitic cirrhosis (P less than 0.01). IR amount was closely related to the TPK in cirrhosis hepatocyte r=0.82597(P less than 0.01). HBV DNA was mainly localized in the nuclei of hepatocyte and pancreatic acinar and islet cells. Immunofluorescent histochemical double-staining showed that HBsAg was partly localized in the IR positive areas of hepatocytes and pancreatic islet cells. CONCLUSION: HBV can invade acinar cells of pancreas and islet cells, which might be a direct cause of insulin-dependent diabetes mellitus-like the disorder and insulin absence after HBV infection. Decrease of IR and TPK might be main cause of noninsulin-dependent diabetes mellitus-like disorder after having hepatitis or posthepatitic cirrhosis. PMID- 15340557 TI - [Establishing DNA chip technique for detecting hepatitis C virus genotypes and primary application]. AB - OBJECTIVE: To establish a new method based DNA chip technique for detecting HCV genotypes. METHODS: Genotyping probes were designed according to the sequence of HCV 5' NCR to generate DNA chip. The probes on DNA chip contains 5 major genotypes and 8 subtypes. The DNA fragment amplified by labeling Cy5 fluorescence was hybridized with DNA chip. RESULTS: Fifty-five out of 65 isolates detected by DNA chip belonged to 1b- DNA sequencing of form a part of the isolates was used as the control. The results of both were completely consistent. CONCLUSION: The method is simple and rapid with high specificity and sensitivity. It can be applied in detection of HCV RNA and genotypes. PMID- 15340558 TI - [Anti-SARS virus activities of different recombinant human interferons in cell culture system]. AB - BACKGROUND: To study the anti-SARS virus activities of different recombinant human interferons on the cell culture system. METHODS: Anti-SARS virus activities of interferons were determined by using CPE inhibition test in human skeletal muscle sarcoma (Rda) cell culture. RESULTS: The average minimum amount of interferon alpha 2b, alpha 1b, beta 1b or omega 1b to inhibit 50% CPE in Rda cell culture was (160.5+/-129.5) IU/ml, (149.0+/-71.7) IU/ml, (69.5+/-61.5) IU/ml, (87.3+/-47.1) IU/ml, respectively or (0.6+/-0.5) ng/ml, (10.6+/-5.1) ng/ml, (3.5+/-3.1) ng/ml, (0.9+/-0.5) ng/ml, respectively. CONCLUSION: All the tested recombinant interferons showed anti-SARS virus activities on the Rda cell culture with different sensitivities. PMID- 15340559 TI - [Development of the cDNA chip for SARS virus and a primary study on the possible molecular mechanism of interferon alpha2b inhibiting the SARS virus replication]. AB - BACKGROUND: To study the molecular mechanism of interferon alpha2b(IFNalpha2b) inhibiting the SARS virus replication. SARS-associated coronavirus (SARS virus) cDNA chip was developed and applied to detect the virus RNA transcription levels in the interferon-treated and untreated cell cultures, and the mechanism of anti SARS virus activity of interferon alpha2b in cell culture system was explored. METHODS: SARS virus cDNA chip was prepared by comparing the published SARS virus genome sequence, and the cDNA chip was used to study the interferon alpha2b function during SARS virus replication. RESULTS: SARS virus cDNA chip was successfully prepared by using PCR method. The results showed that the cDNA chip could be used to detect the viral RNA transcription level. Interferon alpha2b could inhibit almost all the SARS virus gene transcription. An unknown gene at the position 28130-28426 bp, named as U gene, may play an important role during the viral replication. CONCLUSION: A SARS virus whole genome cDNA chip was established. It could be used to study the virus molecular biology and antiviral drug screening. The results also showed that interferon alpha2b could inhibit almost the whole virus gene transcription by using the cDNA chip. PMID- 15340560 TI - [Isolation and identification of SARS virus in Guangdong province]. AB - BACKGROUND: To isolate and identify pathogen of atypical pneumonia in Guangdong. METHODS: Pathogens were isolated from variety of samples collected from atypical pneumonia patient by using MDCK cells, and identified with serological and molecular methods. RESULTS: A novel coronavirus was isolated from patients with atypical pneumonia, from which an RNA fragment of 279 nt was amplified by nested RT-PCR. And sequence assay showed that only 39-65 percent of sequence of the virus was homogenous to known coronavirus, but almost 100% homogenous (with one base exception, 12a to t) to SARS-associated coronavirus isolated from patients outside Guangdong, such as in Beijing, Hong Kong, Taiwan, Germany, Italy and so on. Indirect immunofluorescence test showed a specific antigen-antibody reactivity between the coronavirus and convalescent-phase sera of SARS patients. CONCLUSION: The pathogen of the atypical pneumonia in Guangdong province was a novel type of coronavirus, which could be isolated by using MDCK cells. PMID- 15340561 TI - [Clinical pathology and pathogenesis of severe acute respiratory syndrome]. AB - BACKGROUND: To explore the pathological features and pathogenesis of severe acute respiratory syndrome (SARS) to provide evidence for the clinical treatment and prevention of SARS. METHODS: Pathological features of 2 cases of full autopsy and 4 cases of needle biopsy tissue samples from the patients who died from SARS were studied by light and electron microscopy. The distribution and quantity of lymphocyte subpopulations in the lungs and immune organs from SARS patients were analyzed by immunohistochemistry. The location and semi-quantitative analysis of SARS coronavirus in the tissue specimens were studied by electron microscopy, in situ hybridization and immunohistochemistry. RESULTS: In total of 6 cases, diffuse alveolar damage and alveolar cell proliferation were common. The major pathological changes of 2 autopsy cases of SARS in lung tissues were acute pulmonary interstitial and alveolar exudative inflammation, and 2 autopsy and one biopsy lung tissues showed alveolar hyaline membrane formation. Terminal bronchiolar and alveolar desquamation of lung tissues in one autopsy and 2 biopsy cases were noted. Among 6 cases, 2 biopsy cases presented early pulmonary fibrosis and alveolar organization. Meanwhile, the immune organs, including lymph nodes and spleens from 2 autopsy cases of SARS whose disease courses were less than 12 days showed extensive hemorrhagic necrosis, reactive macrophage/histocyte proliferation, with relative depression of mononuclear and granulocytic clones in the bone marrows. However, spleen and bone marrow biopsy tissue samples from 4 dead SARS cases whose clinical course lasted from 21 to 40 days presented repairing changes. SARS coronaviruses were mainly identified in type I and II alveolar epithelia, macrophages, and endothelia; meanwhile, some renal tubular epithelial cells, cardiomyocytes, mucosal and crypt epithelial cells of gastrointestinal tracts, parenchymal cells in adrenal glands, lymphocytes, testicular epithelial cells and Leydig's cells were also detected by electron microscopy combined with in situ hybridization. The semi-quantitative analysis of lymphocyte subpopulations revealed that the proportion of CD8+ T lymphocytes were about 80% of the total infiltrative inflammatory cells in the pulmonary interstitium, with a few CD4+ lymphocytes CD3+, CD4+, CD8+ or CD20+ lymphocyte subpopulations were obviously decreased and there was imbalance in number and proportion, while CD57+, CD68+, S-100+ and HLA-DR+ cells were relatively increased in lymph nodes and spleens. CONCLUSIONS: Histologically, the pulmonary changes could be divided into acute inflammatory exudative, terminal bronchiolar and alveolar desquamative and proliferative repair stages or types during the pathological process of SARS. SARS coronavirus was found in multi-target cells in vivo, which means that SARS coronavirus might cause multi-organ damages which were predominant in lungs. There were varying degrees of decrease and imbalance in number and proportion of lymphocyte subpopulations in the immune organs of the patients with SARS. However, these changes may be reversible. It was found that cellular immune responses were predominant in the lungs of SARS cases, which might play an important role in getting rid of coronaviruses in infected cells and inducing immune mediated injury. PMID- 15340562 TI - [Predictors associated with clinical deterioration in SARS patients]. AB - BACKGROUND: To study the predictive factors associated with clinical deterioration in SARS patients. METHODS: The clinical data of 60 SARS patients were analyzed by logistic regression and Cox's proportional hazards analysis. RESULTS: In logistic regression models, both older age (P=0.009) and severe lymphopenia (P=0.004) were significant predictors of clinical deterioration. In Cox's proportional hazard models, severe lymphopenia was significant predictor associated with prolongation of stay in hospital. CONCLUSION: Older age and severe lymphopenia seem to be statistically significant for predicting the clinical deterioration in SARS patients. PMID- 15340563 TI - [Characterization of internal genes of two strains of influenza A (H9N2) virus isolated from men]. AB - BACKGROUND: To understand the characteristics of internal genes of two strains of influenza A(H9N2) virus isolated from men and on the basis of these to reveal the origin of these two strains of influenza A(H9N2)virus. METHODS: The target gene was amplified by RT-PCR,the PCR product was ligated with P GEM-T Vector (Promega Company, USA) at 4 degrees, the recombined plasmid was transferred into dH5a bacteria, and the positive colonies were selected and identified with restriction enzyme. Afterwards, they were sent to Liu He Tong Company in Beijing for nucleotide sequencing. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (version 1.03)and Editseq (Version 3.69) softwares. RESULTS: Internal genes of the two strains of H9N2 virus were G9 lineage. There was a slight difference in nucleotide sequence in PA gene between the two strains, whereas another five gene segments were identical to each other. CONCLUSION: The genomes of the two strains of influenza A(H9N2)virus were G9 lineage.They were transmitted to men separately from different avian sources with different characteristics of gene of influenza A(H9N2) virus, respectively. PMID- 15340564 TI - [Detection of Epstein-Barr virus in T cell lymphoma and clinicopathologic analysis]. AB - BACKGROUND: To study the relationship of Epstein-Barr virus (EBV) and T cell lymphoma. METHODS: Sixty cases of T cell lymphomas were examined for the presence of EBV using in situ hybridization for EBV encoded RNA (EBERs). RESULTS: EBERs were detected in tumor cells in 37(69.8%) of 53 cases with peripheral T cell lymphoma, but in none of seven cases of precursor T lymphoblastic lymphoma. The total detected EBERs were 37(61.6%) in 60 cases of T cell lymphomas. By Revised European-American Lymphoma(REAL) classification, EBERs were detected in 2/2 angioimmuno-blastic T cell lymphoma,17/18 angiocentric lymphoma, 4/6 anaplastic large cell lymphoma and 14/27 peripheral T cell lymphoma, unspecified (51.9%). The frequency of EBERs among the extranodal peripheral T cell lymphoma was higher than the nodal (P less than 0.01) there was no significant correlation with the sex, age and clinical stage. CONCLUSIONS: This study indicated that high incidence of EBV was observed in peripheral T cell lymphoma, with predilection for angiocentric lymphoma and extranodal presentation. PMID- 15340565 TI - [Induction of human oral carcinoma by human papillomavirus 16 E6/E7 and TPA]. AB - BACKGROUND: To study the effect of human papillomavirus (HPV) 16 E6/E7 and TPA (12-O-tetradecanog-1-phorbol-13-acetate) on malignant transformation of human embryo oral tissue. METHODS: Recombinant plasmid with HPV 16 E6/E7 was constructed and transfected into human embryo oral tissue. The oral tissue with HPV 16 E6/E7 gene or without the gene was inoculated into the hypophloeodal of right shoulder in scid mice, respectively. The study was conducted in four groups: the first group was the oral tissue transfected plasmid with HPV 16 E6/E7 plus TPA, which were inoculated into 8 scid mice; the second group was only oral tissue transfected with plasmid with HPV 16 E6/E7 into 6 scid mice; the third group was normal oral tissue plus TPA inoculated into 6 scid mice, and the final group was only normal oral tissue inoculated into 5 scid mice. Three days after inoculation, TPA was injected at the left shoulder of the mice once a week. Twelve weeks after inoculation, tumor was found in 7 scid mice from the first group. HPV 16 E6/E7 gene in tumor tissues was analyzed by PCR. RESULTS: The rate of tumor formation was 7/8 in the first group; no tumor was found in the other groups. Pathological diagnosis of the tumor was fibrohistiocytoma. HPV 16 E6/E7 gene was detected by PCR in tumor tissues. CONCLUSION: With the cooperating action of TPA, human oral tissue containing HPV 16 E6/E7 gene could cause malignant transformation in scid mice. PMID- 15340566 TI - [Establishment of primary cultured neurons model of mouse embryo with HSV-1 infection]. AB - BACKGROUND: To establish a neurons model for herpes simplex virus type 1 (HSV-1) infection using cultures of mouse embryo. METHODS: The cortical neuron of 14-16 day old mouse embryo were cultured and infected with HSV-1. Microscopic examination, immunofluorescent test, MTT colorimetric analysis and flowcytometric assay were used to detect the immunofluorescent reaction of normal and infected neurons and astrocytes, respectively. The percentage of the two kinds of cells were measured. RESULTS: The mouse neurons could proliferate well in vitro. The neurons amount met the requirement of cytological experiment after treatment with Ara-C in the tissue culture. HSV-1 could infect the neurons directly, the activity of the infected neurons was obviously reduced. CONCLUSION: HSV-1 could successfully infect the primary cultured neurons. The model may provide a useful approach for studying the pathogenesis of herpes simplex virus encephalitis. PMID- 15340567 TI - [Generation of human recombinant antibody Fab fragment and its IgG to adeno associated virus type II from phage display library]. AB - BACKGROUND: To acquire the recombinant human monoclonal antibodies and IgG to adeno-associated virus type 2 (AAVs-2). METHODS: Construct and pan human Fab antibody library to AAVs-2 was established from normal volunteer donors by using phage display technology and secreted expression in E.Coli system. The positive Fab clones were selected and characterized through ELISA and immunofluorescent assay, and then the heavy and light chain were sequenced. The gene of light chain and heavy chain Fd fragment of recombinant mAb were inserted into baculovirus expression vector pAC-L-Fc and construct expression vectors of intact IgG, then transfected insert sf-9 cell secreted expression in Baculovirus/Insert system. Immunoprecipitation test was used to detect its recognizing region. RESULTS: One clone named AAVs-31 showed positive responses in ELISA and IFA, the Fab was composed of gamma chain and kappa chain IgG was positive in ELISA and IFA. The IgG failed to detect nonassembled or denatured capsid proteins, but recognized the AAVs-2 stock from immunoprecipitation test. CONCLUSION: The authors isolated a clone of Fab and IgG to adeno-associated virus type 2 by phage display technology, they perhaps recognize an epitope which is formed during capsid assembly. PMID- 15340568 TI - [Serial pathologic changes in livers of Tree shrews and Macaca assamensises infected with human Hepatitis B virus]. AB - BACKGROUND: To serially observe the pathologic changes in livers of tree shrews and macaca assamensises infected with HHBV. METHODS: 10 adult tree shrews and 28 macaca assamensises were inoculated with HBV rich human sera. The liver of the animals were regularly biopsied. The liver samples were examined histopathologically by HE staining. Some samples were stained for HBsAg by immunohistochemistry (IH), and HBV DNA by in situ hybridization (ISH). RESULTS: HBsAg in 80% of tree shrews infected with HHBV can be detected by IH, HBV DNA in 50% of those can be found by ISH.The positive rates of HBsAg in macaca assamensises' livers were 25% by IH, none HBV DNA was detected. CONCLUSION: The tree shrew model seems to be applicable for the research of human hepatitis B. PMID- 15340569 TI - [Human anti-HCMV neutralizing Fab antibody generated by phage display library]. AB - BACKGROUND: Cloning recombinant human Fab fragment against HCMV for the purpose of prophylaxis and control of HCMV infection. METHODS: The authors constructed a HCMV phage display library with 2 x 10(6) clones, then used purified HCMV viral lysates to pan the library, then screened by ELISA. RESULTS: Three clones showed positive responses in ELISA, they also showed high specificity in IFA, two of them could neutralize HCMV in neutralizing assays. CONCLUSION: The specific binding of Fab antibodies to HCMV was demonstrated by ELISA, IFA and neutralizing activities. These results provide us the basis for further research of neutralizing recombinant human whole IgG molecule. PMID- 15340570 TI - [Investigation of an outbreak of acute diarrhea caused by Human calicivirus]. AB - BACKGROUND: To survey a diarrhea outbreak in Guangan city and analyze the cause of the disease. METHODS: The population enrolled in the surveillance came from four different settings and was randomly sampled. Stool specimens collected from diarrhea patients were tested ordinarily for enteric bacteria and further examined for viral pathogens with PAGE, ELISA and RT-PCR. RESULTS: In total, 4,567 persons were surveyed, among them 942 had acute diarrhea (prevalence 20.63%). The incidence was higher in rural area (28.6%) than in urban area (19.6%) (chi-square =22.29, P less than 0.005) with a peak in May 10 through 25 four human caliciviruses were detected from stool specimens by ELISA and RT-PCR in specimens from 4 and 1 patients, respectively. CONCLUSION: Human calicivirus probably was the cause of this diarrhea outbreak in Guangan city. PMID- 15340571 TI - [Using recombinant antigens of Hantavirus to study the kinetics of serum IgA, IgG, IgM antibodies in the acute-phase of hemorrhagic fever renal syndrome]. AB - BACKGROUND: To observe the features of serum specific IgA, IgG, IgM antibodies in the acute phase of hemorrhagic fever renal syndrome (HFRS). METHODS: The nucleocapsid (NP) protein and glycoproteins (GP) of Hantavirus were expressed by recombinant baculovirus, and used as ELISA antigens to test 61 serial sera of 14 acute phase HFRS patients. RESULTS: Seoul like virus RNA were detected from 11 of 14 patients. An early and strong IgA, IgG and IgM antibody response to recombinant NP (rNP) was observed in almost all HFRS cases. The titers of antibody to rNP was apparently higher than that to Rgp. In the early stage, titer of IgG antibody elevated most drastically among all the three classes of antibodies to rNP, followed by IgM and IgA antibody responses. The elevation trend of IgM and IgA antibodies to rNP stayed nearly at the same level, but the IgA titers to rNP were apparently higher than that of IgM. Among the antibodies to rGP, IgA changed distinctly greater than IgG. The elevation trend of IgM could be found during first week after the onset, and the titers dropped gradually after the second week. IgM antibodies of one case who was viral RNA positive were not detected at early stage, but IgA titers were high. The only severe case of the 14 patients kept the lower IgA, IgG and IgM during the whole acute phase. CONCLUSION: HFRS patients kept an early and strong humoral response to NP and GPs in acute phase of HFRS.IgA could be used together with IgM to improve the diagnostic accuracy. PMID- 15340572 TI - [Antigencity identification of recombinant hepatitis E virus ORF2 protein expressed in Pichia pastoris]. AB - BACKGROUND: To determine the antigenicity of recombinant hepatitis E virus ORF2 (rHEV ORF2) protein expressed in Pichia pastoris (P. pastoris). METHODS: By using the rHEV ORF2 protein from E.coli as control, an indirect ELISA was adopted to identify the sensitivity, specificity and stability of rHEV ORF2 protein from P. pastoris in detection of HEV IgM and IgG antibody in sera from patients with hepatitis E. The reactivity of the rHEV ORF2 against 5 HEV ORF2 monoclonal antibodies (McAbs) was also tested. RESULTS: The minimum concentration of coated antigen with which HEV IgG could be detected was 12.5 ng/ml, while the highest serum dilution to detect both IgM and IgG antibodies against HEV was 1:5 120. No cross-reaction was found with sera from patients with any other types of hepatitis. The 37 degree C acceleration test showed that the rORF2 was highly stable within 12 months at 4 degrees C. The 5 HEV ORF2 McAbs showed better reaction with the rORF2 from P. pastoris, especially that 4B2, 2E2, whose reaction against the rORF2 were 125 and 25 times respectively higher than that of rORF2 from E.Coli. CONCLUSION: There may be more extensive conformational epitopes in the rHEV ORF2 from P. pastoris. The excellent antigenicity, sensitivity and stability suggest that it can be served as a new candidate antigen for the development of diagnostic reagents of hepatitis E. PMID- 15340573 TI - [Inclusion bodies of human cytomegalovirus are composed of the DNA and immediately early and early antigens of the virus]. AB - BACKGROUND: To study the composition and significance of the inclusion bodies of human cytomegalovirus (HCMV). METHODS: Microdissection of inclusion bodies, PCR and Southern blot were adopted to detect DNA, and immunohistochemistry method and catalyzed signal amplification (CSA) were used to detect the different antigens of HCMV. RESULTS: The inclusion bodies of HCMV were separated from the tissue section of human salivary gland. The fragments amplified by PCR from these dissected inclusion bodies were confirmed to be the DNA of HCMV. With the immunohistochemical method CSA, the immediately early and early antigens of HCMV were detected with monoclonal antibodies DDG9/CCH2, while matrix protein AAC10 was negative in the inclusion bodies. CONCLUSION: The ingredient of inclusion bodies of HCMV included the DNA and the antigens expressed in specific stage of the virus. PMID- 15340574 TI - [Detection of the mutation in HBV polymerase gene by RFLP PCR method in hepatitis B patients treated with lamivudine]. AB - BACKGROUND: To investigate the mutation of HBV polymerase gene in chronic hepatitis B patients treated with lamivudine. METHODS: The restriction-fragment length-polymorphism (RFLP) assay for HBV DNA sequence determination at the codon 528 and 552 in the HBV polymerase gene associated with lamivudine resistance in vitro. HBV DNA samples extracted from sera of 240 patients were subjected to PCR amplification with primer pairs F2/R2 (552), F3/R2 (528). Each PCR product was digested with Nde I or Nla III. RESULTS: Serum HBV DNA mutation was found in 51/240 patients (38/51M552V, 26/38L528M, 13/51M552I) after therapy for 52 weeks. DNA sequence analysis was performed on samples of 3 patients, and the results were consistent with those of RFLP assay. CONCLUSION: The RFLP assay was able to detect the mutation of HBV DNA at codon 552 and 528 which are the principal site of HBV DNA resistant to lamivudine. The specific PCR method for HBV DNA mutation is rapid, simple and specific. PMID- 15340575 TI - [Cut-off period of subclassification and pathological features of severe hepatitis based on clinical and pathological analyses]. AB - BACKGROUND: To explore the cut-off period of subclassification and pathological features of severe hepatitis (SH). METHODS: Based on combined clinical and pathological analyses, the complete clinical and biopsy or autopsy liver tissues data from 196 cases of patients with severe hepatitis were investigated. Meanwhile, proliferative hepatocytes, cholangioepithelia and collagens were identified by a panel of monoclonal antibodies such as those against albumin, cytokeratin 18,19 and collagen I, III with immunohistochemical method. RESULTS: The clinical and pathological analyses indicated the cut-off periods of acute, subacute and chronic SH (ASH,SSH and CSH) were (13.4+/-7.2) d, (77.4+/-69.3) d and (80.5+/-63.2) d, respectively. Among all SH cases, one case of ASH patient presented clinical manifestation and pathological changes of ASH for 21 days, however, one patient with SSH was demonstrated 12 day course by histological examination. The time of cut-off period between ASH and SSH in child cases was shorter than that in adult cases. Histologically, ASH liver tissues showed massive and/or submassive necrosis caused by one attack, with congestive sinusoid frameworks and proliferative cholangioepithelium-like hepatocytes, while SSH liver tissues presented combined fresh and old submassive or massive necrosis caused by multiple attacks, accompanied by obviously proliferative bile ducts and sinusoid framework collapse.However, the pathological changes of CSH showed ASH- or SSH-like lesions on the background of chronic liver injury. CONCLUSION: Our data indicated that the cut-off period between ASH and SSH is in accordance with the Scheme of Viral Hepatitis Prevention and Therapy, China, published in 2000, but excluded a part of child SH cases. In our study, the authors found a few pathological features in ASH and SSH. PMID- 15340576 TI - [Detection of coagulation factor V in patients with severe hepatitis and its clinical significance]. AB - BACKGROUND: To investigate the prognostic significance and role of coagulation factor V (CFV) levels in clinical diagnostic criteria for severe hepatitis. METHODS: The CFV level and prothrombin activity (PTA) were tested by turbidimetry for 129 times in 58 patients with severe hepatitis. Comparative studies and clinical significance of CFV and PTA were analyzed by SPSS and SDAS softwares. RESULTS: 1. The levels of CFV and PTA were 15.3%+/-9.7% and 23.5%+/-10.0%, respectively, at the onset of severe hepatitis. 2. The mortality of severe hepatitis gradually increased with the gradual decrease of CFV or PTA during the most severe stage of the illness (P=0.000). 3. The levels of CFV and PTA decreased continually and rapidly in patients who died but gradually increased in survivors. The decrease or increase of PTA preceded that of CFV on the exacerbation or convalescent stage. 4. Hepatic encephalopathy occurred in 14 cases (24.14%). In 10 cases, it occurred in the terminal stage of the illness, far later than the time of the decrease of CFV. 5. The level of CFV was closely related to PTA (the correlation coefficient was 0.812), the level of CFV was almost consistent with that of PTA. CONCLUSION: 1. The level of CFV is an important prognostic indicator in severe hepatitis and is more specific than PTA. 2. Simultaneous determination of CFV and PTA may be helpful in earlier and more accurate diagnosis of severe hepatitis. 3. Possible use of CFV as one of the criteria for liver transplantation in patients with severe hepatitis should be studied. PMID- 15340577 TI - [Immunological and virological efficacy against HBV chronic infection of the therapeutic vaccine composed of HBV core plus PreS1 in HBV transgenic mice]. AB - BACKGROUND: To investigate the immunological and virological efficacy of the therapeutic vaccine HBV CS1, a recombinant fusion protein which is composed of HBV core aa 1-155 plus PreS1 aa 3-55,against chronic HBV infection. METHODS: HBV transgenic mice were immunized with HBV CS1(5 ug) emulsified in equal volume of complete Freund adjuvant on day 0, followed by a second vaccination with HBV CS1(5 ug) emulsified with incomplete Freund adjuvant on days 21. Mice of control group were mock-vaccinated with PBS plus complete Freund adjuvant/incomplete Freund adjuvant. The splenocytes of individual mouse were subjected to T cell proliferation assays by using 3Hg thymidine, HBsAg and HBV DNA in sera of mice were detected by ELISA and quantitative PCR, respectively. RESULTS: HBV CS1 specific T cell response were induced in mice immunized with HBV CS1, with the titer of HBsAg and the level of HBV DNA decreased significantly after twice immunization with HBV CS1, while the control group almost remained the same. CONCLUSION: HBV CS1 has the immunological and virological efficacy against chronic HBV infection in HBV transgenic mice; HBV CS1 could represent candidate vaccine for further studies on its role as therapeutic vaccine against HBV chronic infection in human. PMID- 15340578 TI - [Hemorheologic changes in patients with liver diseases]. AB - BACKGROUND: To study the variation of the hemorheology and microcirculation in different period of liver diseases. METHODS: Indices for hemorheology, liver function, HBV DNA and transfusion transmitted virus (TTV) DNA were measured in 82 patients with liver diseases and correlative analysis was made. RESULTS: The low shear whole blood viscosity (BV) and RBC aggregation index were significantly higher in hepatitis B group than those in the control group (P less than 0.05). No correlation was found between HBV DNA and indices of hemorheology (P greater than 0.05). The high-shear and low-shear BV and hematocrit (HCT) were significantly lower in decompensated cirrhosis group than those in the control group (P less than 0.05). RBC aggregation index, plasma viscosity (PV) and the low-shear BV were significantly higher in compensated cirrhosis group than those in the control group (P less than 0.05). The high-shear and low-shear BV were significantly higher in TTV positive group than those in the control group. CONCLUSION: There is disturbance of microcirculation in the body of patients with hepatitis B or TTV infection. The blood of patients with compensated cirrhosis is in highly viscose status and in low viscose status in patients with decompensated cirrhosis. TTV seems to be harmful to some degree to the body. The hemorheology should be an index in detecting liver diseases in addition to HBV markers. PMID- 15340579 TI - [Analysis of serological findings and clinical manifestations of TORCH infections in newborns]. AB - BACKGROUND: To search for the serological findings and early clinical manifestations as evidences for prevention and treatment TORCH infections in pregnant women and newborns as early as possible. METHODS: ELASA was performed to screen specific anti-TORCH (Toxoplasma gondii, Cytomegalovirus, Rubella virus, Herpes simplex virus) Ig-M antibodies. RESULTS: Totally 1,554 in-patients who were treated in Neonatal Intensive Care Unit (NICU) of our hospital from January 2000 to January 2003 were retrospectively studied, 48 of them had TORCH infections. Cytomegalovirus (CMV), rubella and herpes simplex virus infections accounted for 52.1%, 33.3% and 14.6%, respectively. None of them had toxoplasma infection. CONCLUSION: TORCH infections can cause multiorgan lesions, such as hearing impairment, hyperbilirubinemias and liver dysfunction, impairment of neurologic system, myocardial impairment, thrombocytopenia, and congenital heart disease.Rubella vaccine inoculation, serological screening during pregnancy and early period of newborn, intervention and treatment in the early period are most important. PMID- 15340580 TI - [Capturing-ELISA for the detection of anti-HCMV-IgM antibody in serum]. AB - BACKGROUND: To develop a capturing-ELISA for the detection of anti-HCMV-IgM antibody in serum. METHODS: The anti-HCMV-IgM antibody was detected in 68 patients with HCMV infection by the capturing-ELISA, and the results were compared with those of indirect ELISA. RESULTS: The specificity and sensitivity of the capturing-ELISA were shown to be significantly higher than those of indirect ELISA, and its results were not affected by RF factor. CONCLUSION: The capturing ELISA is specific, sensitive, convenient and reliable method which may be feasible for clinical use. PMID- 15340582 TI - Recommendations for improved acute pain services: Canadian collaborative acute pain initiative. AB - The Canadian Collaborative Acute Pain Initiative, established in 2002, is a voluntary, multidisciplinary consortium of acute pain health professionals from across Canada whose goal is to improve acute pain management through discussion and consensus. The group met in January 2002 to define strategic areas related to the treatment of acute pain. The areas identified were: the definition of pain; the epidemiology of pain; the concept of an 'ideal' acute pain management service; education; therapeutic options; symptom management; and research and safety. In November 2002, a second meeting was held to develop objectives and recommendations for the management of acute pain based on the defined areas. The outcome of these discussions is summarized in this paper. PMID- 15340581 TI - State of the Art Compendium: Canadian Thoracic Society recommendations for the management of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a common cause of disability and death in Canada. Moreover, morbidity and mortality from COPD continue to rise, and the economic burden is enormous. The main goal of the Canadian Thoracic Society's evidence-based guidelines is to optimize early diagnosis, prevention and management of COPD in Canada. The main message of the guidelines is that COPD is a preventable and treatable disease. Targeted spirometry is strongly recommended to expedite early diagnosis in smokers and former smokers who develop respiratory symptoms, and who are at risk for COPD. Smoking cessation remains the single most effective intervention to reduce the risk of COPD and to slow its progression. Education, especially self-management plans, are key interventions in COPD. Therapy should be escalated on an individual basis in accordance with the increasing severity of symptoms and disability. Long-acting anticholinergics and beta-2-agonist inhalers should be prescribed for patients who remain symptomatic despite short-acting bronchodilator therapy. Inhaled steroids should not be used as first line therapy in COPD, but have a role in preventing exacerbations in patients with more advanced disease who suffer recurrent exacerbations. Acute exacerbations of COPD cause significant morbidity and mortality and should be treated promptly with bronchodilators and a short course of oral steroids; antibiotics should be prescribed for purulent exacerbations. Patients with advanced COPD and respiratory failure require a comprehensive management plan that incorporates structured end-of-life care. Management strategies, consisting of combined modern pharmacotherapy and nonpharmacotherapeutic interventions (eg, pulmonary rehabilitation and exercise training) can effectively improve symptoms, activity levels and quality of life, even in patients with severe COPD. PMID- 15340583 TI - Simulated pain and cervical motion in patients with chronic disorders of the cervical spine. AB - The primary objective of the present study was to determine how simulated severe cervical pain affects cervical motion in patients suffering from two distinct chronic cervical disorders: whiplash (n=25) and degenerative changes (n=25). The second objective was to derive an index that would allow the differentiation of maximal from submaximal performances of cervical range of motion. Patients first performed maximal movement of the head (maximal effort) in each of the six primary directions and then repeated the test as if they were suffering from a much more intense level of pain (submaximal effort). All measurements were repeated within four to seven days. In both groups, there was significant compression of cervical motion during the submaximal effort. This compression was also highly stable on a test-retest basis. In both groups, a significantly higher average coefficient of variation was associated with the imagined pain and it was significantly different between the two clinical groups. In the whiplash group, a logistic regression model allowed the derivation of coefficient of variation based cutoff scores that might, at selected levels of probability and an individual level, identify chronic whiplash patients who intentionally magnify their motion restriction using pain as a cue. However, the relatively small and very stable compression of cervical motion under pain simulation supports the view that the likelihood that chronic whiplash patients are magnifying their restriction of cervical range of motion using pain as a cue is very low. PMID- 15340584 TI - Impact of attending a health care conference in Toronto during the severe acute respiratory syndrome crisis: survey of delegates. AB - OBJECTIVE: To describe the impact on delegates of attending the Canadian Pain Society's annual meeting in Toronto during the severe acute respiratory syndrome (SARS) crisis in May 2003. METHODS: A prospective survey design was used. Surveys were sent to all delegates (n=432) who attended the conference, and 294 delegates responded (68% response rate). The survey was developed to determine the level of concern about travelling to Toronto; the adequacy of screening measures; the level of stress about attending; and the occupational consequences of attending. RESULTS: Fifty per cent of the participants were not concerned about travelling to Toronto, while the other 50% expressed some concern ranging from mild to serious. Concerns included being exposed to SARS and fear of transmitting it to others upon return. Reasons for attending the conference despite concern included a desire for continuing education, decrease in the number of reported SARS cases, and perceived minimal risk. Almost one-half (n=140) felt the screening measures at the conference were adequate, while 4% felt they were inadequate and 9% somewhat adequate. Delegates (n=99) suggested that temperature-taking (32.2%), improved screening vigilance (14.4%), SARS screening forms checked daily (9.1%), strictly controlled entry (8.1%) and adopting hospital screening procedures (7.1%) should have been instituted. CONCLUSION: Health care professionals planning conferences in this era of new respiratory diseases can benefit from understanding the responses of delegates who attended conferences during outbreaks. Clear communication about the potential risks and benefits, as well as instituting full screening precautions, will help to allay concerns. PMID- 15340585 TI - The life adjustment process in chronic pain: psychosocial assessment and clinical implications. AB - Previously, the subjective dimension of suffering from chronic pain has only infrequently been reported in the literature. However, in recent years qualitative psychosocial research has added new perspectives that describe how suffering from chronic pain affects life. The present paper refers to a qualitative study on the subjective experience of women diagnosed with chronic musculoskeletal pain. In a model for life adjustment, turning points and passages are described in three stages. In Stage I, there was increasing pain, disability, and physical and mental exhaustion. Alongside frustration and chaos there was a struggle to restore daily life; therefore, the picture of a healthy future was still intact. Acknowledging that pain may not be temporary marked the transition into Stage II, which was characterized by sorrow and loss, and the picture of the future faded. Once given an explanation of pain (a 'pain diagnosis'), a new understanding developed, with gradual improvement of coping skills. Adjustment eventually progressed into Stage III with the constructive use of past experiences, and competence and control increased. A new picture of the future emerged that could be handled. However, living with pain still required regular maintenance work. The outcome of qualitative investigations, including the one presented here, is discussed along with certain clinical implications. The following clinical implications are reviewed in the present study: distrust from health professionals may obstruct the adjustment process, prolong sick leave and hinder rehabilitation; providing the patient with a diagnosis (eg, an explanation of pain mechanisms) will facilitate life adjustment; acknowledging the individual experience of suffering from chronic pain will make concepts related to pain behaviour, secondary gain and sick role liable to question; and vocational rehabilitation measures should be harmonized with the actual stage of life adjustment. PMID- 15340586 TI - The computer-assisted cognitive/imagery system for use in the management of pain. AB - BACKGROUND: There is growing interest in computer-delivered psychological interventions for a number of clinical conditions, including pain. OBJECTIVES: This study tests the effectiveness of a new computer-delivered pain-management program using a laboratory pain paradigm. METHODS: One hundred twenty undergraduate students were randomly assigned to either the computerized pain management group or the distraction control group. Subjects underwent a cold pressor task and were asked to continuously rate their subjective pain experience. RESULTS: Women receiving the computerized pain management intervention were able to tolerate the cold-pressor task longer than those in the control group. No effect was found for men. Subjective pain ratings were consistently lower during the cold-pressor task for subjects in the computerized pain-management group regardless of sex. Subjects receiving the computerized intervention reported feeling more comfortable and relaxed than control subjects during the cold-pressor task. CONCLUSIONS: Findings indicate that further investigations of the program used in this study are warranted to determine its potential clinical utility and that of similar computerized interventions for pain. PMID- 15340587 TI - Patterns of forensic practice. PMID- 15340588 TI - A challenge for green chemistry: designing molecules that readily dissolve in carbon dioxide. AB - Carbon dioxide is a green yet feeble solvent whose full potential won't be realized until we develop a more thorough understanding of its solvent behavior at the molecular level. Fortunately, advances in molecular modeling coupled with experiments are rapidly improving our understanding of CO(2)'s behavior, permitting design of new, more sustainable "CO(2)-philes". PMID- 15340589 TI - Crystal engineering of the composition of pharmaceutical phases. Do pharmaceutical co-crystals represent a new path to improved medicines? AB - The evolution of crystal engineering into a form of supramolecular synthesis is discussed in the context of problems and opportunities in the pharmaceutical industry. Specifically, it has become clear that a wide array of multiple component pharmaceutical phases, so called pharmaceutical co-crystals, can be rationally designed using crystal engineering, and the strategy affords new intellectual property and enhanced properties for pharmaceutical substances. PMID- 15340590 TI - The carbosilane unit as a stable building block for liquid crystal design: a new class of ferroelectric switching banana-shaped mesogens. AB - New banana shaped liquid crystals with a carbosilane unit at one end were synthesised and depending on the number of Si-atoms either antiferroelectric (AF) or ferroelectric (FE) switching polar smectic C phases have been obtained. PMID- 15340591 TI - Bridge dominated oxidation of a diruthenium 1,3-divinylphenylene complex. AB - A divinylphenylene bridged diruthenium complex constitutes an ensemble of three coupled redox systems. Spectroelectrochemistry provides evidence that the oxidation processes are dominated by the organic bridge. PMID- 15340592 TI - The first utilization of acidic ionic liquid for preparation of tetraarylporphyrins. AB - A phase-separated acidic ionic liquid affords to catalyze a reaction of pyrrole with benzaldehyde in dichloromethane, forming porphyrins in a comparable yield to the Lindsey method and is reusable at least ten times without loss of catalytic activity. PMID- 15340593 TI - Dendron-grafted sulfur-terminated phenyleneethynylene molecular rods and blue luminescence self-assembly with Au nanoparticles. AB - Dendron-grafted phenyleneethynylenes with alpha,omega-disulfur containing groups were newly synthesized and characterized for self-assembly with Au nanoparticles; an intense blue photoluminescence of the composite film was observed. PMID- 15340594 TI - Linking cyano-bridged ladders by azide to form a layered metamagnet. AB - Cyano-bridged corrugated ladder-like chains are connected by mu(1,1)-N(3) (end on, EO) bridges to form a layered coordination polymer, [Mn(N(3))(CH(3)OH)[Cr(phen)(CN)(4)].CH(3)OH (2), which is the first mixed cyano azide bridged compound, and shows metamagnetism below 21.8 K, a temperature much higher than that (3.4 K) of the crossed double chain [Cr(phen)(CN)(4)](2) [Mn(H(2)O)(2)].4H(2)O (1) based on the same building block [Cr(phen)(CN)(4)](-). PMID- 15340595 TI - Functionalisation of a diene-modified hairpin mimic via the Diels-Alder reaction. AB - A highly stable 1,3-butadiene-derived DNA hairpin mimic and its derivatisation via the Diels-Alder reaction with various dienophiles are described. PMID- 15340596 TI - Synthesis and vesicle formation of a nido-carborane cluster lipid for boron neutron capture therapy. AB - The nido-carborane lipid, which has a double-tailed moiety, was synthesized from heptadecanol in 5 steps. Analysis in a transmission electron microscope by negative staining with uranyl acetate showed that the lipid formed a stable vesicle in which calcein was encapsulated. The lipid was incorporated into distearoylphosphatidylcholine (DSPC) liposomes at a very high concentration. PMID- 15340597 TI - An intelligence ink for oxygen. AB - A generic ink formulation is described, comprising semiconductor photocatalyst particles, a brightly-coloured redox dye, a mild reducing agent, a polymer and a solvent, that creates an irreversible, reusable, UV-light-activated, colorimetric indicator or intelligence ink for oxygen. PMID- 15340598 TI - A novel solid-phase equivalent to the triflate group and its application to traceless linking and cross-coupling-release strategies. AB - A solid-phase version of the triflate group has been developed and its use demonstrated in traceless linking of aromatics and cross-coupling release strategies. PMID- 15340599 TI - A 'triflate-like' tetrafluoroarylsulfonate linker for multifunctional solid-phase organic synthesis. AB - An arylsulfonate solid-phase linker is suitable for 'traceless' synthesis and Pd(0) catalyzed cross-coupling reactions. PMID- 15340600 TI - Brookite-supported highly stable gold catalytic system for CO oxidation. AB - A significant enhancement of the gold catalysis stability against sintering has been achieved using brookite as a catalytic support. PMID- 15340601 TI - Hexagonal, hollow, aluminium-containing ZSM-5 tubes prepared from mesoporous silica templates. AB - Hexagonal hollow ZSM-5 tubes were synthesized using mesoporous silica with a worm like morphology as the template. A new method for aluminium incorporation during the hydrothermal synthesis step was developed. PMID- 15340602 TI - A simple and highly efficient P,O-type ligand for Suzuki-Miyaura cross-coupling of aryl halides. AB - A simple and efficient hemilabile-type phosphine ligand, found to be highly effective in Suzuki-Miyaura cross-coupling of aryl chlorides with generally low Pd-catalyst loading (0.05%), was prepared in one step based on an economically attractive approach from commercially available benzamide starting material. PMID- 15340603 TI - Solvent free synthesis of polyaniline-clay nanocomposites from mechanochemically intercalated anilinium fluoride. AB - Nanocomposites consisting of conducting polyaniline and clay minerals were successfully synthesized from mechanochemically intercalated anilinium fluoride; the nanocomposites prepared by the mechanochemical intercalation method contained much more polyaniline in the clay layers than those prepared by a conventional solution method. PMID- 15340604 TI - Direct arylation of aromatic C-H bonds catalyzed by Cp*Ir complexes. AB - The C-H bond of benzene was directly arylated by reaction with aryl iodides in the presence of a catalytic amount of a pentamethylcyclopentadienyliridium complex and potassium tert-butoxide. PMID- 15340606 TI - A highly efficient gold/silver-catalyzed addition of arenes to imines. AB - A highly efficient imino Friedel-Crafts type addition of arenes to imines is developed by using a combination of gold and silver catalysts; various amino acid derivatives are generated by this method. PMID- 15340605 TI - First catalytic asymmetric synthesis of beta-amino-beta-polyfluoroalkyl ketones via proline-catalysed direct asymmetric carbon-carbon bond formation reaction of polyfluoroalkylated aldimines. AB - Proline-catalysed direct asymmetric carbon-carbon bond formation reaction of polyfluoroalkylated aldimines with acetone afforded the corresponding beta-(p methoxyphenyl)amino-beta-polyfluoroalkyl ketones in high enantioselectivities (up to 98% ee). PMID- 15340607 TI - Palladium catalyzed arylation reactions of meta photocycloadducts. AB - We describe the first application of a Heck arylation reaction being performed on a meta photocycloadduct. The resulting arylation/fragmentation process leads to the formation of a bridged bicyclic ketone and constitutes a dramatic increase in molecular complexity after just two synthetic operations. PMID- 15340608 TI - CuI-catalyzed coupling reaction of aryl halides with terminal alkynes in the absence of palladium and phosphine. AB - CuI/N,N-dimethylglycine-catalyzed coupling reaction of aryl halides with terminal alkynes is carried out in DMF at 100 degrees C to provide the corresponding coupling products in good to excellent yields with a great diversity. PMID- 15340610 TI - Ionic liquids as catalytic green solvents for cracking reactions. AB - We demonstrate the use of room-temperature ionic liquids as catalysts and solvents for the cracking reaction of alkoxypropanes, eliminating the need for a volatile organic solvent and hazardous catalyst disposal. PMID- 15340609 TI - Looking glass inhibitors: L-DMDP, a more potent and specific inhibitor of alpha glucosidases than the enantiomeric natural product DMDP. AB - L-DMDP, prepared from D-gulonolactone, is a highly specific inhibitor of a number of plant and mammalian alpha-glucosidases [between 2 and 4 orders of magnitude more potent than the enantiomeric natural product DMDP] but is not an inhibitor of bacterial and yeast alpha-glucosidases. Additionally N-butyl-DMDP is a potent inhibitor of ceramide-specific glucosyltransferase but N-butyl-L-DMDP shows no inhibition. PMID- 15340611 TI - Template-assisted self assembly of two lipophilic polyion aggregates derived from sodium tetraphenyl imidodiphosphinate-complexes containing sodium ions in four different coordination environments. AB - The molecular structures of two lipophilic polyion aggregates derived from tetraphenyl imidodiphosphinate are described: [Na(crown ether)][MNa(2)[Ph(2)P(O)NP(O)Ph(2)](4)] with crown ether = 15-crown-5 for 1and benzo-15-crown-5 for (M = Na(+) for 1 and Na(H(2)O)(+) for 2). PMID- 15340612 TI - Conversion of [Pt(SRf)2(PPh2 -n(C6F5)n+ 1)2]( n = 0 or 1, Rf=C6HF4-4) through carbon-fluorine bond activation to [Pt(SRf)2(1,2-C6F4(SRf)-(PPh2))] and chiral [Pt(SRf)2(1,2-C6F4(SRf)(PPh(C6F5)))]. AB - Treatment of trans-[PtCl(2)(PPh(2 - n)(C(6)F(5))(n + 1))(2)](n = 0 or 1) with Pb(SC(6)HF(4)-4)(2) yields a mixture of monometallic cis/trans [Pt(SC(6)HF(4) 4)(2)(PPh(2 - n)(C(6)F(5))(n + 1))(2)], thiolate-bridged bimetallic cis/trans [Pt(2)(mu-SC(6)HF(4)-4)(2)(SC(6)HF(4)-4)(2)(PPh(2 - n)(C(6)F(5))(n + 1))(2)] and [Pt(SC(6)HF(4)-4)(2)(1,2-C(6)F(4)(SC(6)HF(4)-4)(PPh(2 - n)(C(6)F(5))(n))]. PMID- 15340613 TI - Ion/molecule reactions of the protonated serine octamer. AB - The protonated homochiral octamer of serine exchanges all 33 of its labile hydrogens with CH(3)OD and undergoes ligand switching reactions with amines in a quadrupole ion trap mass spectrometer. PMID- 15340614 TI - A charge transfer-type fluorescent molecular sensor that "lights up" in the visible upon hydrogen bond-assisted complexation of anions. AB - A charge transfer-type fluorescent molecular sensor consisting of a bisamidopyridine receptor and two styryl base chromophores shows H(2)PO(4)(-) and acetate-enhanced fluorescence due to the conversion of weak intramolecular hydrogen bonds into strong ones in the host-guest ensemble. PMID- 15340615 TI - Direct oxidation of methane to acetic acid catalyzed by Pd2+ and Cu2+ in the presence of molecular oxygen. AB - Methane is catalytically converted primarily to acetic acid in concentrated sulfuric acid using a combination of Pd(2+) and Cu(2+) in the presence of oxygen. PMID- 15340616 TI - Initialisation in RAFT-mediated polymerisation of methyl acrylate. AB - Inhibition in Reversible Addition-Fragmentation chain Transfer (RAFT)-mediated polymerisations is currently a controversial issue; here we provide evidence that the slow "propagation" of the initiating and leaving group radicals during the early part (the period of consumption of the initial RAFT agent) of methyl acrylate RAFT-mediated polymerisation has characteristics similar to inhibition. PMID- 15340617 TI - Phosphoramidic acid monoesters as phosphorylating agents: steric effects and reluctance to form monomeric metaphosphate intermediates. AB - The formation of phosphate diesters (RO)(2)P(X)OH (R = Et, Pr(i) or Pr(i)(2)CH) by phosphorylation of ROH with ROP(X)(NPr(i)(2))OH is insensitive to steric effects when X = S but not when X = O; this is consistent with a unimolecular mechanism and a thiometaphosphate (ROPOS) intermediate when X = S but a bimolecular S(N)2(P) mechanism when X = O. PMID- 15340618 TI - Vanadyl complexes bearing bi- and triphenolate chelate ligands: highly active ethylene polymerisation procatalysts. AB - Vanadyl procatalysts bearing chelating aryloxide ligands are found to polymerise ethylene, in the presence of dimethylaluminium chloride, with (for this metal) exceptionally high activities. PMID- 15340619 TI - Bis(benzimidazole)amine vanadium catalysts for olefin polymerisation and co polymerisation: thermally robust, single-site catalysts activated by simple alkylaluminium reagents. AB - Vanadium complexes containing bis(benzimidazole)amine ligands, upon activation by simple alkylaluminium reagents, give unusually robust, single-site, catalysts for olefin polymerisation/co-polymerisation. PMID- 15340620 TI - Water addition to a two-electron mixed-valence bimetallic center. AB - Water adds to the two-electron mixed-valence Ir(0,II)(2) core of Ir(2)(tfepma)(3)Cl(2)(tfepma = MeN[P(OCH(2)CF(3))(2)](2)) to cleanly generate an Ir(I,III)(2) hydride. Dehydrohalogenation across the Ir-Ir bond returns the complex to an Ir(0,II)(2) species. PMID- 15340621 TI - One-pot oligosaccharide synthesis: reactivity tuning by post-synthetic modification of aglycon. AB - Post-synthetic aglycon modification of substituted aryl thioglycosides led to building blocks with multiple levels of anomeric reactivities, which can be used in rapid assembly of oligosaccharides in one-pot syntheses. PMID- 15340622 TI - Radical addition of nitrones to acrylates mediated by SmI2: asymmetric synthesis of gamma-amino acids employing carbohydrate-based chiral auxiliaries. AB - Alkyl nitrones possessing N-substituted sugars as chiral auxiliaries were found to effectively undergo an SmI(2)-mediated radical addition to n-butyl acrylate affording gamma-amino acid derivatives with high diastereomeric control. PMID- 15340623 TI - Super-hydrophobic tin oxide nanoflowers. AB - Super-hydrophobic 3D SnO(2) flowers with nanoporous petals were produced from the 3D Sn nanoflowers using a controlled shape-preserving thermal oxidation process. PMID- 15340624 TI - (Bu3Sn)2-TBAF: a new combination reagent for the reduction and deuteration of aryl bromides and iodides. AB - The combination of (Bu(3)Sn)(2) and TBAF has been shown to reduce aromatic bromides and iodides in excellent yields under mild conditions. When the residual water in TBAF is exchanged for D(2)O, the halogen is replaced by a deuterium atom. PMID- 15340625 TI - KF-Silica as a stationary phase for the chromatographic removal of tin residues from organic compounds. AB - Through the simple expedient of using a mixture of KF and silica as the stationary phase in column chromatography, levels of organotin impurities from tributyltin hydride mediated reductions have been reduced from stoichiometric levels to approximately 30 ppm. PMID- 15340626 TI - A new reaction: lead(IV) acetate-mediated oxidative fragmentation of homoallylic alcohols. AB - Treatment of a homoallylic alcohol with lead(IV) acetate in refluxing benzene for 2 hours results in an oxidative cleavage to give a carbonyl compound and an allylic acetate with high yield. PMID- 15340627 TI - Electrochemical reduction of an imidazolium cation: a convenient preparation of imidazol-2-ylidenes and their observation in an ionic liquid. AB - 1,3-Bis(2,4,6-trimethylphenyl)imidazolium chloride is reduced electrochemically and chemically to produce a nucleophilic carbene, namely 1,3-bis(2,4,6 trimethylphenyl)imidazol-2-ylidene. The carbene was also shown to be compatible with and persistent in the ionic liquid tetradecyl(trihexyl)phosphonium chloride. PMID- 15340628 TI - A new synthetic route to produce metal zeolites with subnanometric magnetic clusters. AB - By using a Co layered silicate as a source of Co and SiO(2), Co-Beta and Co-ZSM-5 zeolites with tetrahedrically coordinated cobalt were synthesized. Subnanometric clusters of Co can be produced and a magnetic material with superparamagnetic paramagnetic phase transition is obtained. PMID- 15340629 TI - Palladium catalysed cross-coupling reactions of silylamines. AB - The palladium-catalysed formation of C-N bonds to produce a range of aryl amines in supercritical carbon dioxide is reported; carbamic acid formation is avoided in part by the use of N-silylamines as surrogates for the free amine. PMID- 15340630 TI - Preparation of primary amine-modified gold nanoparticles and their transfection ability into cultivated cells. AB - Cationic gold nanoparticles, prepared by reducing HAuCl(4) with NaBH(4) in the presence of 2-aminoethanethiol, formed a complex with plasmid DNA, and could be transfected into cultivated cells. PMID- 15340631 TI - Thiostannylation of arynes with stannyl sulfides: synthesis and reaction of 2 (arylthio)arylstannanes. AB - Arynes were found to insert into a sulfur-tin sigma-bond of stannyl sulfides to give a variety of 2-(arylthio)arylstannanes, whose carbon-tin bonds were applicable to further transformations. PMID- 15340632 TI - Darmstadtium carbonyl and carbide resemble platinum carbonyl and carbide. AB - DsC and DsCO are calculated to be very similar to PtC and PtCO, suggesting that even the latest 6d elements resemble the corresponding 5d elements chemically. PMID- 15340633 TI - Rhodium catalysed tandem conjugate addition-protonation: an enantioselective synthesis of 2-substituted succinic esters. AB - The rhodium catalysed addition of potassium trifluoro(organo)borates to dimethyl itaconate generates an intermediate complex which on protonation provides enantioenriched succinic esters. PMID- 15340634 TI - High-quality free-standing and oriented periodic mesoporous organosilica films grown without a solid substrate at the air-water interface. AB - In this communication, we report the first synthesis of high-quality free standing and oriented periodic mesoporous organosilica (PMO) films grown without a solid substrate, by surfactant templating at the air-water interface. PMID- 15340635 TI - Super-long continuous Ni nanowires encapsulated in carbon nanotubes. AB - Super-long continuous Ni-filled carbon nanotubes were synthesized by the chemical vapor deposition (CVD) method with cloth-like single-walled carbon nanotube (SWNT) raw soot produced by the arc-discharge method as catalyst; the Ni nanowires inside the carbon nanotubes are single crystals, with an average diameter of 40 nm and up to tens of micrometres in length. PMID- 15340636 TI - A highly efficient heterogeneous catalytic system for Heck reactions with a palladium colloid layer reduced in situ in the channel of mesoporous silica materials. AB - A new catalyst, Pd-SBA, was prepared by the introduction of an Si-H function into the channel of SBA-15 mesoporous materials resulting in a highly dispersed metal colloid layer on the pore walls of the support material, creating one of the most active heterogeneous catalysts for Heck coupling reactions. PMID- 15340637 TI - Adjustable cavity for host-guest recognition in cofacial bis-porphyrinic tweezer. AB - The synthesis of a cofacial bis-porphyrinic tweezer bearing a tris-anthracenic spacer is reported. Its behavior as host has been evidenced as well as the ability of its cavity to adjust to guests of various sizes. PMID- 15340638 TI - Application of a diphosphinidenecyclobutene ligand in the solvent-free copper catalysed amination reactions of aryl halides. AB - 1,2-Diphenyl-3,4-bis[(2,4,6-tri-tert-butylphenyl)phosphinidene]cyclobutene was used as an additive ligand in copper-catalysed amination reactions of halobenzenes with amines in the presence of base to afford the corresponding secondary or tertiary amines in good to excellent yields. PMID- 15340639 TI - Complementary hydrogen-bonding between thymidine-based low molecular-weight gelator and polynucleotide in organic media. AB - We have found that aggregation mode and gelation property of thymidine-based organogelator are affected by the addition of its complementary polynucleotide in organic media. PMID- 15340640 TI - Preparation and properties of an ordered, uniform 0.9 nm Ag array assembled in a nanoporous VSB-1 by a simple soft chemical method. AB - An ordered, homogeneously distributed and uniform 0.9 nm Ag array, which exhibits an unusual UV-vis surface plasmon resonance absorption peak at 355 nm with the largest blue-shift position and controllable intensity, has been successfully prepared by a simple soft chemical approach using the nanoporous VSB-1 as the template. PMID- 15340641 TI - Ionic liquid assisted preparation of nanostructured TiO2 particles. AB - Anatase-containing nanostructured TiO(2) particles with high surface area have been synthesized using a water immiscible room temperature ionic liquid (1-butyl 3-methylimidazolium hexafluorophosphate) as an effective additional solvent by the sol-gel method at low temperature. PMID- 15340642 TI - Alpha,beta-unsaturated imines from titanium hydroamination and functionalization by rhodium C-H activation. AB - Titanium pyrrolyl complexes are effective catalysts for the synthesis of alpha,beta-unsaturated imines, which can be further functionalized using Rh catalyzed alkene or alkyne insertion into the beta-C-H bond. PMID- 15340643 TI - The first oligopeptide derivative of 1'-aminoferrocene-1-carboxylic acid shows helical chirality with antiparallel strands. AB - The unnatural organometallic amino acid 1'-aminoferrocene-1-carboxylic acid (Fca) induces a turn structure in a tetrapeptide with anti-parallel strands which is stabilized by two intra-molecular hydrogen bonds in the solid state and in solution. PMID- 15340644 TI - A single-enzyme, two-step, one-pot synthesis of N-substituted imidazole derivatives containing a glucose branch via combined acylation/Michael addition reaction. AB - Combined regioselective acylation/Michael addition reaction catalyzed by alkaline protease from Bacillus subtilis in anhydrous pyridine for synthesis of N substituted imidazole derivatives containing a glucose branch via a novel single enzyme, two-step, one-pot procedure is reported. PMID- 15340645 TI - Replacing HCl by solid acids in industrial processes: synthesis of diamino diphenyl methane (DADPM) for producing polyurethanes. AB - Delaminated zeolite catalysts allow replacement of HCl in the production of DADPM: they offer higher activity and catalyst life than conventional zeolites, and their structured external surface controls product selectivity. PMID- 15340646 TI - Cloning, sequencing and expression of GroEL-like protein gene of Vibrio parahaemolyticus. AB - Vibrio parahaemolyticus is an important enteropathogen in regions where much seafood is consumed. Substantial quantity of GroEL-like protein is produced during the heat shock of V. parahaemolyticus and located in periplasmic and extracellular fractions. In this study, the GroEL-like protein gene of this pathogen was cloned and sequenced and its properties were analyzed. The open reading frame consisted of 1647 bp, encoded a 57.6-kDa GroEL-like protein of 548 amino acids. The amino acid sequence, hydrophobicity and antigenic pattern of V. parahaemolyticus GroEL-like protein were similar to the GroEL proteins of other vibrios, Escherichia coli and Salmonella enterica Typhi. The groEL-like gene of V. parahaemolyticus was cloned into the pQE-30 expression vector and the expression of a His-tagged GroEL-like protein was rapidly induced by isopropyl beta-D-thiogalactopyranoside, largely in an insoluble form. The results of this study facilitate the study of the functions of the GroEL-like protein of V. parahaemolyticus. PMID- 15340647 TI - Evidence for arylamine N-acetyltransferase activity in Klebsiella pneumoniae. AB - Arylamine N-acetyltransferase (NAT) enzymes have been found in laboratory animals, humans, microorganisms (fungi, bacteria and parasites), and in plants. But the characteristics of NAT from Klebsiella pneumoniae are not clear. NAT activities with p-aminobenzoic acid (PABA) and 2-aminofluorene (AF) as substrates were examined in the cytosol of K. pneumoniae. NAT activity (N-acetylation of substrates) was determined using an acetyl coenzyme A recycling assay and high performance liquid chromatography for determining the amounts of acetylated or non-acetylated PABA or AF. NAT activities from a number of K. pneumoniae isolates were found to be 0.72 +/- 0.08 nmol/min/mg protein for AF, and 0.49 +/- 0.04 nmol/min/mg protein for PABA. The kinetic parameters of apparent Michaelis constant (Km) and maximum velocity (Vmax) obtained were 2.92 +/- 0.48 mM and 7.89 +/- 0.82 nmol/min/mg protein, respectively, for AF and 2.42 +/- 0.28 mM and 9.87 +/- 0.64 nmol/min/mg protein, respectively, for PABA. The optimal pH value for the NAT activity was 7.0 for AF and PABA. The optimal temperature for NAT activity was 37 degrees C for both substrates. The NAT activity was inhibited by 50% with 0.25 mM iodoacetamide, and by more than 90% at 1.0 mM. Among a series of divalent cations and salts, Cu2+ and Zn2+ were the most potent inhibitors of NAT activity. PMID- 15340648 TI - An open study of inactivated hepatitis A vaccine (VAQTA) in Taiwanese healthy adult volunteers: safety, tolerability, and immunogenicity. AB - The safety, tolerability and immunogenicity of inactivated hepatitis A vaccine (VAQTA, Merck and Co. Inc., West Point, PA, USA) were investigated in 28 seronegative healthy adult volunteers. The age range was 25-35 years, and the mean age was 29 years. Two doses of the vaccine, each containing 50 U of hepatitis A virus antigen, were administered into the deltoid region 24 weeks apart. No serious vaccine-related adverse reactions were reported. Four weeks after the first dose, the geometric mean titer (GMT) was 150 mIU/mL, and the seroconversion rate was 100%. Twenty eight weeks after the first dose (4 weeks following the second dose), the GMT was 4576 mIU/mL. This study demonstrated that VAQTA is safe and highly immunogenic in healthy young adults in Taiwan. PMID- 15340649 TI - Kikuchi-Fujimoto disease in children: clinical features and disease course. AB - Kikuchi-Fujimoto disease (KFD) is a rare cause of cervical lymphadenitis which mostly affects Asian women but is sometimes observed in the pediatric population. This study analyzed the clinical manifestations and disease course in children with KFD. Retrospective chart review and telephone interview were used to collect data for 13 children (8 boys and 5 girls) with a diagnosis of KFD from January 1988 to January 2003. Involvement of the posterior cervical lymph nodes was found in 12 patients, leukopenia in 9, and all patients had C-reactive protein less than 5 mg/dL. A high antinuclear antibody titer was associated with a more protracted and complicated course. Five of the 13 patients had new symptoms compatible with an autoimmune process during follow-up, with neurological symptoms the most common. In conclusion, the risk of evolution into an autoimmune syndrome in pediatric KFD patients is high, and careful long-term observation is mandatory. PMID- 15340650 TI - Clinical course of children of human immunodeficiency virus-infected mothers in Taiwan. AB - The objective of this study was to describe the clinical courses of infants born to human immunodeficiency virus (HIV)-infected mothers in Taiwan. Eleven children, including 1 set of twins, born to 10 HIV-infected mothers were included in the study. HIV antibodies were assayed and HIV-1 polymerase chain reaction (HIV-PCR) or virus cultures were performed; HIV infection was established when there were at least 2 separate positive results of HIV-PCR or culture. Three sets of primers detecting LTR-gag, pol and env were used. The viral load of HIV RNA was measured and used as an indicator of the treatment response. Two of the children were HIV-infected and received combination therapy, including 2 kinds of nucleoside analogue reverse transcriptase inhibitors plus 1 protease inhibitor. Neither of these children exhibited HIV-related symptoms or signs during the study period. Both mothers of the infected children were Taiwanese and their HIV infection status was not known during pregnancy. In contrast, HIV infection was found in early pregnancy in the 4 women living in Taiwan who were from other countries, all of whom received prophylactic therapy. The other 4 mothers who did not transmit the infection to the infant were Taiwanese, 3 of whom were known to be HIV-seropositive during pregnancy. Based on these results, the vertical rate of transmission was 18% (2/11). Early detection of HIV-infected pregnancy is vital to reduce the incidence of HIV-infected births. PMID- 15340651 TI - Community-acquired brain abscess in Taiwan: etiology and probable source of infection. AB - Brain abscess is a life-threatening infection caused by spread from infected parameningeal or remote foci. Historically, streptococci have been the predominant organisms reported while brain abscess metastatic from liver abscess caused by Klebsiella pneumoniae has been a more recent emerging problem. This study retrospectively analyzed the characteristics of community-acquired brain abscess admitted during an 11-year period. There were 17 men and 7 women with age from 20 to 82 years (median, 41 years). The most common source of infection was liver abscess, followed by otitic infection and sinusitis. The classic triad of fever, headache and focal neurologic deficit was noted in only 25% of cases. Spread of the abscess to multiple lobes was common (n = 6). The most commonly identified organisms were Streptococcus spp. (n = 7) and K. pneumoniae (n = 5). All 5 cases of K. pneumoniae brain abscess also had concomitant pyogenic liver abscess and 4 of them had diabetes mellitus. In this study, brain abscess was common in young patients and in patients with diabetes mellitus. In Taiwan, Streptococcus spp. and K. pneumoniae are leading etiologies for community acquired brain abscess. Liver abscess is the most likely source of K. pneumoniae brain abscess. PMID- 15340652 TI - Species distribution and fluconazole susceptibility of Candida clinical isolates in a medical center in 2002. AB - Fluconazole disk-diffusion susceptibility was evaluated in 230 blood isolates and 344 non-blood clinical isolates of Candida spp. collected in 2002 at National Taiwan University Hospital. Up to 93.5% of blood isolates were susceptible to fluconazole, 3% were susceptible dose-dependent, and 3.5% were resistant. The minimum inhibitory concentrations at which 50% of tested isolates were inhibited (MIC50) of fluconazole against Candida blood isolates were highest for Candida glabrata (5 microg/mL), followed by Candida tropicalis (2.4 microg/mL), Candida albicans (2.4 microg/mL), and Candida parapsilosis (0.41 microg/mL). C. glabrata had less fluconazole-susceptible strains (76.7%) than C. albicans (98.2%), C. tropicalis (98%) and C. parapsilosis (93.8%) [p<0.05]. The proportions of fluconazole resistance in the non-blood isolates of C. albicans, C. glabrata and C. parapsilosis were similar to those of the blood isolates. However, the proportions of fluconazole resistance in the non-blood isolates of C. tropicalis surpassed those of the blood isolates (14.7% vs 2%, p<0.05). Comparison of species distribution of Candida blood isolates obtained in 2002 to those in 1981 2000 demonstrated that C. albicans remained the leading pathogen, and the proportion of C. albicans in blood isolates was lowest in 1996 (38%) and did not change significantly thereafter. However, the proportion of C. tropicalis increased from 14% during 1981-1993 to 22-23% during 1996-2002. Overall, the MIC50, MIC90 and the proportion of Candida blood isolates with fluconazole resistance remained stable during 1994-2002. PMID- 15340653 TI - Proficiency of determination of vancomycin susceptibility in enterococci by clinical laboratories in Taiwan. AB - Eighty clinical microbiology laboratories in Taiwan were evaluated for proficiency in the determination of vancomycin susceptibility of enterococci. Each laboratory was given 1 vancomycin-sensitive isolate and 3 vancomycin resistant enterococci (VRE) isolates to determine the levels of vancomycin resistance. Among a total of 240 tests performed, 153 (63.8%) correctly determined the levels of vancomycin resistance of the survey isolates. Seventy eight (98%) of the 80 laboratories accurately identified the high level of vancomycin resistant isolates [Enterococcus faecalis; minimum inhibitory concentration (MIC) >256 microg/mL]. Seventy two laboratories (90%) correctly determined the level of vancomycin resistance of another VRE with a vancomycin MIC of 64 microg/mL. Only 3 of the 80 laboratories correctly determined the intermediate-level vancomycin resistant isolates (Enterococcus casseliflavus; MIC = 8 microg/mL). Eight laboratories reported the vancomycin-susceptible isolate as being vancomycin-resistant or of intermediate susceptibility. This survey demonstrated that clinical microbiology laboratories in Taiwan are proficient in detecting high-level but not low-level VRE, suggesting a need to improve their proficiency in VRE detection. PMID- 15340654 TI - Giant cell arteritis with CD8+ instead of CD4+ T lymphocytes as the predominant infiltrating cells in a young woman. AB - Giant cell arteritis is rarely reported in people aged less than 50 years. We report a case of giant cell arteritis in a woman who developed symptoms of dizziness, headache, bilateral sensorineural hearing impairment, and had 1 episode of transient left hemiparesis before the age of 30. Carotid angiography showed multiple segmental narrowing in cranial vessels. Subsequently, at the age of 31, she had weight loss and developed a fever. Chest radiograph revealed mediastinal widening, and chest computed tomography revealed dilated pulmonary arteries and veins. Coronary angiography and aortography showed irregular narrowing of the descending aorta and multiple stenosis, with aneurysmal dilatation involving the proximal and distal coronary, pulmonary and mesenteric arteries. Multinucleated giant cells and predominant CD8+ T lymphocyte infiltration were noted in a left temporal artery biopsy specimen. The patient's age and the finding of dilated pulmonary veins and prominent CD8+ T lymphocytes in the biopsy specimen suggest that this case was a distinct form of systemic giant cell arteritis. PMID- 15340655 TI - Primary adrenal insufficiency in patients with acquired immunodeficiency syndrome: report of four cases. AB - Adrenal insufficiency, though rare, is a well-known complication of acquired immunodeficiency syndrome (AIDS). Over the past 9 years at the National Taiwan University Hospital, 4 patients were diagnosed with primary adrenal insufficiency among 854 non-hemophiliac patients with human immunodeficiency virus infection, with an incidence of 0.19 per 100 patient-years (95% confidence interval, 0.17 0.21 per 100 patient-years). All of the 4 patients were severely immunosuppressed and had been diagnosed with at least 1 AIDS-defining opportunistic illness ever reported to produce adrenal insufficiency in AIDS patients. In 1 of the patients, fluconazole was considered contributory to the development of adrenal insufficiency after treatment at a daily dose of 800 mg for 68 days. PMID- 15340656 TI - Non-septicemic Burkholderia pseudomallei liver abscess in a young man. AB - Melioidosis, caused by Burkholderia pseudomallei, has been increasingly recognized in Taiwan recently. Its isolation in liver abscess is rare compared to pulmonary melioidosis. We report a case of liver abscess due to B. pseudomallei in an immunocompetent 27-year-old male soldier admitted due to fever, sore throat and mild non-productive cough for 1 week. Physical examination was unremarkable except for congestion of the pharyngeal wall, moderate enlargement of the tonsils without pus coating, and palpable tender lymphadenopathy over bilateral submental regions. Antibiotic treatment with cefazolin 1 g every 8 hours intravenously was given without response. Left flank pain, followed by right flank pain associated with epigastric tenderness developed. Sonography and computed tomography scan of the abdomen demonstrated liver abscess. Aspiration of the liver abscess was performed and abscess culture yielded B. pseudomallei. Treatment with ceftazidime 2 g every 8 hours intravenously (4 weeks' duration) followed by oral regimens of amoxicillin-clavulanate was given. The patient was free of symptoms at 8 months' follow-up. Early awareness and definite diagnosis as well as institution of proper antimicrobial agents are imperative for successful treatment of melioidosis. PMID- 15340657 TI - Recent advances in the management of primary breast cancers. AB - The current concept of breast cancer treatment arises from Fisher's theory that operable breast cancer has distant micrometastasis at its very early stages. Since it is the presence of systemic diseases or micrometastasis that determines the final outcome, variation in local treatment would not affect survival. Fisher's theory led to a change in local treatment, from Halsted's radical mastectomy to breast-conserving therapy (BCT), and the introduction of adjuvant systemic treatment. As part of the job of surgery is replaced by radiation therapy in local control, the efficacy and side effects of radiation should be carefully monitored. The recently published results of 20-year follow-up in 2 important studies confirm that BCT achieves equal survival compared to mastectomy in women with early breast cancers, even after all causes of mortality have been considered. The introduction of sentinel lymph node biopsy has further decreased the adverse impact of breast cancer treatment on women. As variation in local control does not affect survival, more efforts are being put into developing adjuvant systemic treatment with curative intent. Adjuvant chemotherapy has been demonstrated to substantially affect the survival of women with early breast cancers. It is now apparent from numerous studies that adjuvant therapy improves survival in all subgroups of women with early breast cancer, although the absolute benefit varies depending on axillary lymph node status, tumor size, and other prognostic factors. This article reviews recent advances in the management of primary breast cancer, including: long-term follow-up after BCT; side effects of radiation therapy in BCT; post-mastectomy radiotherapy; sentinel node biopsy; adjuvant hormone therapy; and chemotherapy, including new strategies such as the incorporation of taxanes, dose-dense chemotherapy schedules, and the use of aromatase inhibitors in place of, or in addition to, tamoxifen. PMID- 15340658 TI - Comparison of the anti-proliferation and apoptosis-induction activities of sulindac, celecoxib, curcumin, and nifedipine in mismatch repair-deficient cell lines. AB - BACKGROUND AND PURPOSE: The adenomatous polyposis coli (APC) and mismatch repair (MMR) pathways are both involved in the tumorigenesis of hereditary colorectal cancers. Chemoprevention focuses on the APC pathway in the absence of information concerning MMR targets. This study compared the anticancer effects of sulindac, celecoxib, curcumin, and nifedipine in MMR-deficient cell lines, in order to determine the most appropriate chemopreventive agent for long-term use in patients with hereditary colorectal cancer. METHODS: Five human colorectal cell lines (SW480, HCT116, LoVo, SW48, and HCT15) and an endometrial cancer cell line (HEC-1-A) were used for susceptibility testing. Tests included assays for growth inhibition, cell-cycle arrest, and apoptosis. RESULTS: Sulindac, celecoxib, curcumin, and nifedipine all displayed dose- and time-dependent anti proliferation activities. Celecoxib was the most effective anti-proliferative agent, and increased the G0/G1 phase proportion in the cell cycle after treatment more significantly than the other agents in all cell lines. Curcumin displayed a more potent apoptosis-inducing activity than the other agents in treated cells. CONCLUSIONS: The tested drugs were effective against colorectal and endometrial cancer cell lines. Celecoxib is more potent with fewer side effects than sulindac. Nifedipine's observed chemopreventive efficacy may complement its known therapeutic application in patients with hypertension. PMID- 15340659 TI - Risk factors for wound infection after cholecystectomy. AB - BACKGROUND AND PURPOSE: Surgical site infection (SSI) after cholecystectomy is a common problem. The aim of this study was to identify the possible risk factors for the development of SSI. METHODS: 545 consecutive patients who received open (125) or laparoscopic (420) cholecystectomy due to gallbladder disease during the years 1998 to 2000 were included in the study. Potential risk factors including clinical features, biochemical data, and operative types were analyzed by univariate and multivariate analysis. RESULTS: The overall incidence of SSI was 4.4% (24/545). The wound complication rate was significantly lower in the laparoscopic group than in the open group (1.4% vs 14.4%, respectively). Factors associated with SSI found by univariate analysis (p < 0.05) included age, gender, acute cholecystitis, white blood cell count, serum albumin, blood glucose and bilirubin level, type of surgery, operative time and positive bile culture. Stepwise logistic regression analysis showed that abnormal blood glucose [odds ratio (OR), 4.7; 95% confidence interval (CI), 1.6 to 13.5], positive bile culture (OR, 3.5; 95% CI, 1.2 to 10.4), and open cholecystectomy (OR, 4.3; 95% CI, 1.3 to 13.6) were the most significant predictors of SSI. CONCLUSION: Poor control of diabetes mellitus before surgery, positive bile culture and open cholecystectomy significantly increased the rate of SSI. These findings indicate that better control of diabetes mellitus, and appropriate selection of surgical procedure and antibiotic regimen in the management of high-risk patients may reduce the incidence of postoperative SSI. PMID- 15340660 TI - Safety and immunogenicity of heptavalent pneumococcal conjugate vaccine in Taiwanese infants. AB - BACKGROUND AND PURPOSE: The prevalence of antibiotic-resistant pneumococci has increased markedly in Taiwan since the 1990s. The prevention of pneumococcal infection by effective vaccination is an urgent need. The purpose of this study was to assess the safety and immunogenicity of heptavalent pneumococcal conjugate vaccine (serotypes 4, 6B, 9V, 14, 18C, 19F, 23F) in Taiwanese infants. METHODS: Sixty healthy Taiwanese infants received heptavalent pneumococcal conjugate vaccine at 2, 4 and 6 months of age in an open-label, non-comparative trial. Routine diphtheria-tetanus-pertussis vaccines were concomitantly administered. Side effects were recorded and the antibody titer to each of the 7 pneumococcal serotypes was measured before vaccination and 1 month after the third vaccination. RESULTS: Mild to moderate local reactions at the injection site were the common findings (17 to 30%) after vaccination but were not correlated with the number of vaccinations. Fever, restlessness, fussiness, and loss of appetite were the most common systemic reactions (22 to 53%). After 3 doses of vaccine, the geometric mean concentration of immunoglobulin G showed a significant rise (p < 0.001) to all 7 pneumococcal serotypes. More than 95% of infants had antibody titer > or = 0.15 microg/mL of all serotypes, and 93% (serotype 23F) to 100% (serotypes 4 and 19F) of infants achieved an antibody titer > pr = 1 microg/mL. CONCLUSION: Inoculation with the heptavalent pneumococcal conjugate vaccine including the prevalent serotypes in Taiwan is safe and effective in preventing pneumococcal disease in Taiwanese children. PMID- 15340661 TI - A comparative study of etanercept plus methotrexate and methotrexate alone in Taiwanese patients with active rheumatoid arthritis: a 12-week, double-blind, randomized, placebo-controlled study. AB - BACKGROUND AND PURPOSE: Etanercept (Enbrel), a recombinant tumor necrosis factor receptor fusion protein, has been shown to be effective in the treatment of patients with rheumatoid arthritis (RA). The purpose of this study was to compare the efficacy and safety of etanercept in combination with methotrexate (MTX) and MTX alone in Taiwanese patients with active RA. METHODS: In this double-blind study, 58 patients with active RA who were maintained on MTX therapy at a stable dose of 12.5 to 20 mg per week for 4 weeks were randomized to receive either etanercept 25 mg (n = 29) or placebo (n = 29) by subcutaneous injection twice weekly over a period of 12 weeks. The primary endpoint was the reduction of tender and swollen joint counts by 20% (ACR 20), 50% (ACR 50), and 70% (ACR 70) as determined by the American College of Rheumatology criteria at the 12th week. RESULTS: The addition of etanercept to MTX resulted in a greater reduction in the number of tender (7.00 vs 2.45, p = 0.012) and swollen joints (8.55 vs 3.86, p = 0.017), and in serum levels of C-reactive protein (1.26 mg/dL vs 0.45 mg/dL, p = 0.014) compared to MTX alone after 12 weeks of therapy. In addition, the global assessment of disease activity by both physicians and patients, duration of morning stiffness, pain visual analog scale score, and Health Assessment Questionnaire were all improved by etanercept plus MTX therapy. Results for the overall improvement in disease activity assessed by ACR 20 (90% vs 34%), ACR 50 (66% vs 10%) and ACR 70 (24% vs 0%) all favored the etanercept plus MTX group. However, the adverse events were comparable between the 2 treatment groups. CONCLUSION: Etanercept in combination with MTX was well tolerated and provided significantly more clinical benefit than MTX alone in Taiwanese patients with active RA. PMID- 15340662 TI - The physiological impact of wearing an N95 mask during hemodialysis as a precaution against SARS in patients with end-stage renal disease. AB - BACKGROUND AND PURPOSE: Most patients with end-stage renal disease (ERSD) visiting our hospital for hemodialysis treatment during the SARS outbreak wore an N95 mask. Data on the physiological stress imposed by the wearing of N95 masks remains limited. This study investigated the physiological impact of wearing an N95 mask during hemodialysis (HD) on patients with ESRD. METHODS: ESRD patients who received regular HD at National Taiwan University Hospital between April to June 2003 were enrolled. Each patient wore a new N95 mask (3M Model 8210) during HD (4 hours). Vital signs, clinical symptoms and arterial blood gas measured before and at the end of HD were compared. RESULTS: Thirty nine patients (23 men; mean age, 57.2 years) were recruited for participation in the study. Seventy percent of the patients showed a reduction in partial pressure of oxygen (PaO2), and 19% developed various degrees of hypoxemia. Wearing an N95 mask significantly reduced the PaO2 level (101.7 +/- 12.6 to 92.7 +/- 15.8 mm Hg, p = 0.006), increased the respiratory rate (16.8 +/- 2.8 to 18.8 +/- 2.7/min, p < 0.001), and increased the occurrence of chest discomfort (3 to 11 patients, p = 0.014) and respiratory distress (1 to 17 patients, p < 0.001). Baseline PaO2 level was the only significant predictor of the magnitude of PaO2 reduction (p < 0.001). CONCLUSION: Wearing an N95 mask for 4 hours during HD significantly reduced PaO2 and increased respiratory adverse effects in ESRD patients. PMID- 15340663 TI - Bardet-Biedl syndrome initially presenting as McKusick-Kaufman syndrome. AB - Bardet-Biedl syndrome (BBS) and McKusick-Kaufman syndrome (MKKS) are rare congenital disorders of autosomal recessive inheritance. Because of the phenotypic overlap of both syndromes, including hydrometrocolpos (HMC) and postaxial polydactyly (PAP) in the neonatal stage, the potential for diagnostic confusion exists. A case of BBS with the initial presentation of MKKS is reported. MKKS was diagnosed during the neonatal period based on the classical findings of HMC together with vaginal atresia and PAP. However, follow-up examination after the age of 2 years revealed additional clinical features consistent with BBS, including mental retardation, obesity, and retinitis pigmentosa. A rehabilitation program was undertaken for the problems of moderate motor delay and slurred speech. MKKS may be considered as a variant of BBS. Careful monitoring for the complications of BBS including ophthalmologic, neurologic, and urologic assessments should be performed in patients with MKKS. PMID- 15340664 TI - Alveolar hydatid disease causing total occlusion of the inferior vena cava. AB - Alveolar hydatid disease is a malignancy-like parasitic disease. It is invasive, metastatic, and almost always lethal if left untreated. A case of alveolar hyatid disease presenting with total of the inferior vena cava is reported. This 28-year old man was referred for elevation of obstructive jaundice. He was a lama and had recently arrived in Taiwan after touring temples in Nepal, India, and Singapore. Computed tomography showed calcified mass which occupied the right hepatic lobe with extension to the left hepatic lobe. The inferior vena cava was occluded and stricture of biliary tree and portal vein at the hilum was also noted. The patient was treated successfully with palliative resection combined with postoperative albendazole. With increasing travel and immigration, clinicians will be more likely to encounter this rare disease, and thus should be able to recognize its symptoms. PMID- 15340665 TI - Successful pregnancy after preimplantation genetic diagnosis in a female with Robertsonian translocation. AB - Preimplantation genetic diagnosis (PGD) is an alternative option for couples with chromosome abnormalities. A 34-year-old woman with balanced Robertsonian translocation [(45, XX, der(13; 14)(q10; q10)] requested PGD due to recurrent spontaneous abortion. Embryos of good quality were biopsied on day 3 post-oocyte retrieval. The aspirated blastomeres were fixed and analyzed using fluorescence in situ hybridization. In the first cycle, 2 unaffected embryos were transferred back without success. No unaffected embryo was available in the second cycle. On day 5 in the third cycle, 2 unaffected embryos were transferred resulting in a twin pregnancy. Amniocentesis confirmed the diagnosis. At the gestational age of 35 weeks, 2 healthy girls were born via cesarean section. Postnatal physical examination found no evidence of major abnormalities. PMID- 15340666 TI - Isolated gastric varices due to focal splenic vein stenosis. AB - Left-sided portal hypertension due to splenic vein stenosis is a very rare disease. We report a case of this condition in a 21-year-old woman who suffered from a first episode of tarry stool passage with fresh blood vomiting. Panendoscopy showed isolated gastric varices while sonography showed a normal liver but the presence of splenomegaly with prominent collateral circulations. Further imaging studies, including abdominal computed tomography, splenoportography and percutaneous transhepatic portography, revealed a focal stenotic proximal splenic vein resulting in left-sided portal hypertension. The collateral circulation ran from the short gastric veins via the left gastric veins into the main portal vein. The intraportal venous pressure was within normal limits. Splenectomy was performed and near normal wedge liver biopsy pathology confirmed non-cirrhotic extrahepatic portal hypertension. The patient had no further variceal bleeding after surgery. PMID- 15340667 TI - Use of a versatile transpositional flap in the surgical treatment of axillary hidradenitis suppurativa. AB - Axillary hidradenitis suppurativa is a chronic recurrent disorder characterized by abscessing inflammation, fistulating sinus tracts, and scarring. This is not only a medical disease, but also a complicated problem necessitating surgical intervention. While various management strategies have been reported, the results are frequently unsatisfactory. Primary closure of the defect after radical excision is often impossible and results in poor wound healing. Skin graft on the soft tissue defect often results in obvious scar contracture and limitation of movement. Local muscle flap can fill in the defect but the range of adduction of the arm will be limited by its muscle bulk. Free flap transfer is a choice of management but this technique calls for a trained team, laborious execution, expensive instruments and plenty of time. We describe the use of transpositional fasciocutaneous flap, which can provide a reliable flap of variable size of skin and soft tissue coverage with good elastic properties. The technique is easily adapted to the reconstruction of resultant defects. This technique was used to transfer 8 transpositional fasciocutaneous flaps in 7 patients for the closure of axillary defects resulting from radical excision of chronic hidradenitis suppurativa. No flap complication or disease recurrence was observed during 2 years of follow-up. The technique had satisfactory esthetic and functional results as well as low donor site morbidity. PMID- 15340668 TI - Charcoal hemoperfusion in an elderly man with life-threatening adverse reactions due to poor metabolism of phenytoin. AB - A 77-year-old man presented with dizziness and ataxia after 7-day treatment of phenytoin 100 mg 3 times daily for prophylaxis of post-traumatic seizure. Thrombocytopenia and hematuria were found incipiently and supportive measures were employed. Owing to extremely slow metabolism of phenytoin in this patient, the period of exposure to phenytoin overdose was prolonged, resulting in delayed hypersensitivity syndrome. Neutropenia and fever developed and thus antibiotics and granulocyte-colony stimulating factor were administered. Although charcoal hemoperfusion is generally not applied in cases of phenytoin overdose, this method was successfully used to enhance the removal of phenytoin in our patient. PMID- 15340669 TI - [The law and the habit of smoking]. PMID- 15340670 TI - [The postgraduate program of the Heart Institute of the School of Medicine, University of Sao Paulo]. PMID- 15340671 TI - [Rescue percutaneous coronary intervention after fibrinolysis]. PMID- 15340672 TI - Angiographic morphologic characteristics in pulmonary atresia with intact ventricular septum. AB - OBJECTIVE: To determine the morphological and functional changes in the right ventricle in pulmonary atresia with intact ventricular septum (PAIVS) for assessing the candidates for the different therapeutic procedures currently available. METHODS: Thirty-one patients underwent cineangiocardiographic study with axial projections. Their ages ranged from 1 to 50 days (x = 9.6), and 28 of them were studied during the first month of life. The statistical analysis comprised the following: chi-square test, Kruskal-Wallis test for the mean and standard deviation, multiple regression, and the 95% confidence interval (95%CI) was calculated. The significance level adopted was alpha < or =0.05. RESULTS: The patients were divided into 3 groups according to the angiographic morphology of the right ventricle (RV): group A -- tripartite RV (n=16); group B -- bipartite RV (n=9); and group C -- unipartite RV (n=6). The diameter of the tricuspid valve was 10.28 +/- 2.67 mm (A); 7.82 +/- 3.41 (B); and 5.27 +/- 0.57 (C) (P=0.0005). Pulmonary atresia was of the valvular type in all group A patients and of the infundibular type in all group C patients (P<0.0001). Coronary-cavitary connections were rare (2/16) in group A patients, but occurred in all group C patients (P=0.0006), with retrograde opacity of the aorta (flow from the RV to the aorta) in 2 group A patients and in all group C patients (P=0.0003). Three patients (2 in group C and 1 in group A) had right-ventricular-dependent coronary circulation. Isolated moderate/severe tricuspid regurgitation showed a tendency towards being more frequent in group A (P=0.0525). The angle of the ductus arteriosus with the descending aorta was as follows: 104.06 +/- 8.98 in group A; 79.17 +/- 33.08 in group B; and 39.0 +/- 6.52 in group C (P=0.0016). The correlation between the diameter of the tricuspid valve and the angle of the ductus arteriosus with the descending aorta was 0.6568 (P=0.0002). CONCLUSION: Because of the heterogeneity of the morphology of the RV in patients with pulmonary atresia with intact ventricular septum, knowledge about all these data is required for selecting candidates for the different therapeutic procedures. PMID- 15340673 TI - Evaluation of systolic, diastolic, and pulse pressure as risk factors for severe coronary arteriosclerotic disease in women with unstable angina non-ST-elevation acute myocardial infarction. AB - OBJECTIVE: To evaluate pressures assessed at the aortic root as risk factors for severe atherosclerotic coronary heart disease in women with unstable angina/compatible clinical history associated with increase in cardiac enzymes (total CPK and CK-MB) 2 times greater than the standard value used in the hospital, with the absence of new Q waves on the electrocardiogram (UA/NSTEMI). METHODS: Five hundred and ninety-three female patients with clinical diagnosis of UA/NSTEMI underwent cinecoronariography from March 1993 to August 2001, and the risk factors for CHD were studied. During examination the pressures, at the aortic root, and coronary obstructions were visually assessed by 2 interventional cardiologists, and those stenosis over 70% were considered severe. RESULTS: Eight one per cent of the population was white and 18.3% was black. Mean age was 59.2+/ 11.2 years, and it was significantly higher in patients with severe coronary lesions: 61.9 +/- 10.8 years versus 56.4 +/- 10.8 years; smoking, diabetes mellitus and climacteric were more frequent in patients with CHD. The average mean arterial pressure and mean systolic blood pressure was the same in both groups, however, average left ventricle diastolic pressure (17.6 +/- 8.7 x 15.1 +/- 8.1, p=0.001), and aortic pulse pressure were significantly greater in patients with CHD (75.5 +/- 22 x 70 +/- 19, p=0.002), while average aortic diastolic pressure was significantly greater in patients without CHD (79.8 +/- 16 x 75.3 +/- 17.5, p=0.003). In the multivariated analysis, pulse pressure > 80 mmHg and systolic blood pressure > 165 were independently associated with severe CHD with odds ratio of 2.12 and 2.09, p<0.05, respectively. CONCLUSION: CHD is associated with increased pulse pressure and lower diastolic blood pressure in women with UA/NSTEMI. Although average systolic blood pressure has not been associated with CHD in this population, dichotomized values of pulse pressure > 80 mmHg and systolic blood pressure > 165 mmHG determined risk two times greater of severe coronary disease. PMID- 15340674 TI - A comparison of rescue and primary percutaneous coronary interventions for acute myocardial infarction: a multicenter registry report of 9,371 patients. AB - OBJECTIVE: To perform a comparative analysis of in-hospital results obtained from AMI patients who underwent rescue or primary PTCA. METHODS: From the Brazilian Interventional National Registry (CENIC), we selected all consecutive patients who underwent a percutaneous coronary intervention for myocardial infarction (< 24 hours), between 1997 and 2000, analyzing those undergoing a rescue (n=840) or a primary (n=8,531) procedure, and comparing their in-hospital results. RESULTS: Rescue patients were significantly younger males with anterior wall infarctions, associated with left ventricular dysfunction, but had less multivessel disease, compared with those treated with primary intervention. Coronary stents were implanted in at similar rates (56.9% vs. 54.9%; P=0.283). Procedural success were lower for rescue cases (88.1% vs. 91.2%; P<0.001), with higher mortality (7.4% vs. 5.6%; P=0.034), compared with the primary intervention group; target vessel revascularization (< or =0.5%), emergency bypass surgery (< or =0.3%) and reinfarction (< or =2.6%) rates were similar for both strategies. Multivariate analysis identified the rescue procedure as a predictor of in-hospital death [OR(CI=95%) = 1.60 (1.17-2.19); P=0.003]. CONCLUSION: Patients who underwent a rescue coronary intervention had higher in-hospital death rates compared with those who underwent a primary coronary intervention. PMID- 15340675 TI - Surgical treatment for intermittent claudication in patients who do not improve with clinical treatment. AB - OBJECTIVE: To study the results obtained with surgical treatment of patients with intermittent claudication (IC) who did not clinically improve with conservative treatment, accompanied by a long follow-up (average 6 years). METHODS: From January 1992 to January 2002, 26 patients treated surgically in a group of 1380 IC patient, representing 1.88% of the total. RESULTS: Sixteen patients did not experience walking limitations after the surgery. Nine patients improved, however, with some degree of limitation. No intraoperative mortalities occurred. Three patients experienced thrombosis of the treated artery 6, 48, and 60 months after the procedure and started to suffer IC with onset at the same distances as before the surgery. During the long-term follow-up, we observed a mortality rate of 23.0% due to myocardial infarctions (4 patients), renal insufficiency (1 patient), and cerebral infarction (1 patient). Two patients underwent coronary bypasses 2 and 4 years after the vascular surgery, and one underwent coronary angioplasty after 3 years of follow-up. The mean follow-up was 73 months. CONCLUSION: In our study, the results from surgical treatment of IC brought about a lasting regression of the ischemic conditions in a significant number of patients, with excellent patency rates (88.4%). We conclude that this is a good alternative for select patients, with low rates of complications and positive long-term results. PMID- 15340676 TI - Concordance of the scoring system for controlling the serum levels of cholesterol and fat. AB - OBJECTIVE: To evaluate the comprehension of the scoring system for controlling the serum levels of cholesterol and fat as an instrument of dietary intervention for hypercholesterolemic patients. METHODS: The study comprised 153 individuals of both sexes, with ages ranging from 20 to 65 years, who sought medical care in a hospital or in a basic health unit. They were divided into the following 3 groups: hypercholesterolemic individuals, individuals with no diagnosis of hypercholesterolemia, and health care professionals. A 24-hour recall was used with each individual, who applied the cholesterol/saturated-fat index to their diets, consulting the scoring system. The researcher confirmed the calculation of each participant, considering her calculation as the a reference pattern. Understanding of the system was assessed through comparison between the reference pattern and the data obtained by the interviewees, using the intraclass correlation coefficient. RESULTS: The patients without a diagnosis of hypercholesterolemia obtained a high correlation in all meals. The morning and afternoon snacks and supper were the meals with the greatest correlation (r = 1). In hypercholesterolemic individuals, supper was the meal with the greatest concordance with the reference score (r = 1), and the health care professionals obtained a correlation coefficient of 1 in all meals, except lunch (r = 0.99). CONCLUSION: The scoring system was fast, simple, and easy to be understood and accepted by the population studied. PMID- 15340677 TI - The posterior ventricular branches of the coronary arteries in the human heart. AB - OBJECTIVE: To describe the trajectory of the posterior ventricular branches of the coronary arteries in the epicardial adipose tissue, and to propose a new criterion for analyzing the distribution of these branches, according to the traditional classification, to determine the predominance or balance of the coronary arteries in the arterial supply to the heart. METHODS: Forty hearts obtained from autopsies of adults were dissected and fixed in a formol solution. The posterior face of each ventricle was divided into 8 approximately equal areas for the morphological classification of the coronary circulation. The following 3 traditional types were considered: A) right dominance, B) balanced, and C) left dominance. The number, diameter, and terminal areas of the posterior ventricular branches in the epicardium were analyzed. RESULTS: The following branches and respective frequencies were found: left marginal branch -- 100%; posterior left ventricular branches: lateral -- 75%, intermediate -- 82.5%, and medial -- 87.5%; interventricular posterior branch -- 95%; posterior right ventricular branch: medial -- 40%, intermediate -- 32.5%, and lateral -- 40%; posterior diagonal branch of the right ventricle -- 17.5%; right marginal branch -- 95%. In regard to dominance, the following values were found: dominance of the right coronary artery -- 62.5%; balanced type -- 25%; and dominance of the left coronary artery - 12.5%. CONCLUSION: The method adopted allowed a more precise classification of the types of coronary artery distribution found. The right coronary artery dominance type was the most prevalent, followed by the balanced type and the left coronary artery dominance. PMID- 15340678 TI - Vascular color Doppler ultrasound for assessing renovascular hypertension: accuracy of the direct technique for assessing the renal arteries. AB - OBJECTIVE: To assess the accuracy of vascular color Doppler ultrasound as compared with digital subtraction arteriography for identifying hemodynamically significant renal artery stenoses. METHODS: One hundred and thirty-seven renal arteries from 69 adult patients suspected of having renovascular arterial hypertension were prospectively studied with ultrasound. The results obtained were compared in a double-blind manner with those obtained on digital subtraction arteriography, and the following parameters were calculated according to previously defined criteria: sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy. RESULTS: The inconclusive results (7 arteries) were excluded. The comparison of the 2 methods in regard to the 130 remaining arteries showed concordant results for 116 (89.2%) arteries and discordant results for 14 (10.8%) arteries. The sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy of vascular color Doppler ultrasound were, respectively, 95.33%, 88.14%, 89.86%, 94.55%, and 91.94%. CONCLUSION: A good correlation between the 2 examinations was observed in the evaluation of the hemodynamically significant renal artery stenoses, making vascular color Doppler ultrasound a noninvasive method useful for selecting patients with suspected renovascular hypertension. PMID- 15340679 TI - Chest pain due to right atrial compression caused by a thymolipoma. PMID- 15340680 TI - Mitochondrial hypertrophic cardiomyopathy associated with Wolff-Parkinson-White syndrome. PMID- 15340681 TI - [Clinico-radiographic correlation: case 5/2004 -- 16-year-old with Ebstein's anomaly. Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 15340682 TI - [Heart failure with preserved systolic function]. PMID- 15340683 TI - Mapping OI-4, a gene conferring resistance to Oidium neolycopersici and originating from Lycopersicon peruvianum LA2172, requires multi-allelic, single locus markers. AB - Lycopersicon peruvianum LA2172 is completely resistant to Oidium neolycopersici, the causal agent of tomato powdery mildew. Despite the large genetic distance between the cultivated tomato and L. peruvianum, fertile F1 hybrids of L. esculentum cv. Money maker x L.peruvianum LA2172 were produced, and a pseudo-F2 population was generated by mating F, half-sibs. The disease tests on the pseudo F2 population and two BC,families showed that the resistance in LA2172 is governed by one dominant gene, designated as 01-4. In the pseudo-F2 population, distorted segregation was observed, and multi-allelic, single-locus markers were used to display different marker-allele configurations per locus. Para-meters for both distortion and linkage between genetic loci were determined by maximum likelihood estimation, and the necessity of using multi-allelic, single-locus markers was illustrated. Finally, a genetic linkage map of chromosome 6 around the 01-4 locus was constructed by using the pseudo-F2 population. PMID- 15340684 TI - Genome-specific primer sets for starch biosynthesis genes in wheat. AB - Common wheat (Triticum aestivum L.,2n=6x=42) is an allohexaploid composed of three closely related genomes, designated A, B, and D. Genetic analysis in wheat is complicated, as most genes are present in triplicated sets located in the same chromosomal regions of homoeologous chromosomes. The goal of this report was to use genomic information gathered from wheat-rice sequence comparison to develop genome-specific primer sets for five genes involved in starch biosynthesis. Intron locations in wheat were inferred through the alignment of wheat cDNA sequences with rice genomic sequence.Exon-anchored primers, which amplify across introns,allowed the sequencing of introns from the three genomes for each gene. Sequence variation within introns among the three wheat genomes provided the basis for genome-specific primer design. For three genes, ADP-glucose pyrophosphorylase (Agp-L), sucrose transporter (SUT),and waxy (Wx), genome specific primer sets were developed for all three genomes. Genome-specific primers were developed for two of the three genomes for Agp-S and starch synthase I (Ssl). Thus, 13 of 15 possible genome-specific primer sets were developed using this strategy. Seven genome-specific primer combinations were used to amplify alleles in hexaploid wheat lines for sequence comparison. Three single nucleotide polymorphisms(SNPs) were identified in a comparison of 5,093 bp among a minimum of ten wheat accessions. Two of theseSNPs could be converted into cleaved amplified polymorphism sequence (CAPS) markers. Our results indicated that the design of genome-specific primer sets using intron-based sequence differences has a high probability of success, while the identification of polymorphism among alleles within a genome may be a challenge. PMID- 15340685 TI - Characterisation and transferability of apple SSRs to two European pear F1 populations. AB - European pear (Pyrus communis L.) is among the important fruit species for which only few genetic studies have been carried out. Available evidence indicates that simple sequence repeats (SSR) are very useful as molecular markers because they are codominant, highly polymorphic, abundant and reproducible. The present paper reports more than 100 apple SSR markers in two populations of European pear; a total of 41 SSR markers were then positioned on a genetic linkage map of the cross 'Passe Crassane' x 'Harrow Sweet' and 31 in the map 'Abbe Fetel' x 'Max Red Bartlett'. Syntenic relationships between pear and apple maps have been considered for the chromosomes carrying two or more SSR markers. The alignment among the two maps supports the colinearity of the two genomes with respect both to identification and to orientation of the linkage groups. PMID- 15340686 TI - Dissemination of the highly expressed Bx7 glutenin subunit (Glu-B1al allele) in wheat as revealed by novel PCR markers and RP-HPLC. AB - Increased expression of the high molecular weight glutenin subunit (HMW-GS) Bx7 is associated with improved dough strength of wheat (Triticum aestivum L.) flour. Several cultivars and landraces of widely different genetic backgrounds from around the world have now been found to contain this so-called 'over-expressing' allelic form of the Bx7 subunit encoded by Glu-B1al. Using three methods of identification, SDS-PAGE, RP-HPLC and PCR marker analysis, as well as pedigree information, we have traced the distribution and source of this allele from a Uruguayan landrace, Americano 44D, in the mid-nineteenth century. Results are supported by knowledge of the movement of wheat lines with migrants. All cultivars possessing the Glu-B1al allele can be identified by the following attributes: (1) the elution of the By sub-unit peak before the Dx sub-unit peak by RP-HPLC, (2) high expression levels of Bx7 (>39% Mol% Bx), (3) a 43 bp insertion in the matrix-attachment region (MAR) upstream of the gene promoter relative to Bx7 and an 18 bp nucleotide duplication in the coding region of the gene. Evidence is presented indicating that these 18 and 43 bp sequence insertions are not causal for the high expression levels of Bx7 as they were also found to be present in a small number of hexaploid species, including Chinese Spring, and species expressing Glu-B1ak and Glu-B1a alleles. In addition, these sequence inserts were found in different isolates of the tetraploid wheat, T. turgidum, indicating that these insertion/deletion events occurred prior to hexaploidization. PMID- 15340687 TI - Sequence tagged microsatellites for the Xgwm533 locus provide new diagnostic markers to select for the presence of stem rust resistance gene Sr2 in bread wheat ( Triticum aestivum L.). AB - The stem rust resistance gene Sr2 has provided durable broad-spectrum, adult plant resistance to the fungal pathogen Puccinia graminis Pers. f. sp. tritici throughout wheat-growing regions of the world for more than 50 years. The ability to select for Sr2 in wheat breeding programs was recently improved by the identification of a tightly linked microsatellite marker gwm533. This marker typically amplifies a 120-bp polymerase chain reaction fragment from wheat lines carrying Sr2. In instances where the 120-bp fragment is not associated with the presence of Sr2, DNA sequence analysis has shown that a second allele was amplified, differing in the structure of the microsatellite repeat. To discriminate this allelic homoplasy (alleles identical in size, but not identical by descent), sequence-tagged microsatellites (STM) markers were developed for the Xgwm533 locus. These markers were shown to be diagnostic for the presence of Sr2 in a wide range of germplasm, representative of all major wheat varieties historically grown in Australia. The STMs will be particularly useful for marker assisted selection in Southern Australian breeding programs, where the use of the marker gwm533 is often precluded by the presence of the non- Sr2-associated 120 bp allele in the pedigree of current breeding germplasm. The STMs also revealed a high incidence of previously undetected allelic homoplasy at the Xgwm533 locus and may have broader utility in genetic research and breeding, as this locus is also reported to be strongly associated with a major gene conferring resistance to Fusarium head blight. PMID- 15340688 TI - Comparative mapping of ZYMV resistances in cucumber (Cucumis sativus L.) and melon (Cucumis melo L.). AB - Zucchini yellow mosaic virus (ZYMV) routinely causes significant losses in cucumber ( Cucumis sativus L.) and melon ( Cucumis melo L.). ZYMV resistances from the cucumber population 'TMG1' and the melon plant introduction (PI) 414723 show different modes of inheritance and their genetic relationships are unknown. We used molecular markers tightly linked to ZYMV resistances from cucumber and melon for comparative mapping. A 5-kb genomic region (YCZ-5) cosegregating with the zym locus of cucumber was cloned and sequenced to reveal single nucleotide polymorphisms and indels distinguishing alleles from ZYMV-resistant (TMG1) and susceptible (Straight 8) cucumbers. A low-copy region of the YCZ-5 clone was hybridized to bacterial artificial chromosome (BAC) clones of melon and a 180-kb contig assembled. One end of this melon contig was mapped in cucumber and cosegregated with ZYMV resistance, demonstrating that physically linked regions in melon show genetic linkage in cucumber. However the YCZ-5 region segregated independently of ZYMV resistance loci in two melon families. These results establish that these sources of ZYMV resistances from cucumber TMG1 and melon PI414723 are likely non-syntenic. PMID- 15340689 TI - Genetic analysis of agronomic and quality traits in mustard (Brassica juncea). AB - To develop an efficient mustard (Brassica juncea) breeding programme, a better knowledge of the genetic control and relationships of the main selected characters is needed. Thus, doubled haploid (DH) lines were evaluated over 2 years in the field. Days to flowering, plant height, thousand-seed weight, fatty acid composition, seed oil content, sinigrin, gluconapin and total glucosinolate contents were determined in the DH population. The influence of seed coat colour was estimated. Results showed significant differences between yellow and brown seeds only for oil and fatty acid contents. Molecular analysis revealed that seed coat colour is associated with two Mendelian trait loci: Bjc1 [on linkage group (LG) 3] and Bjc2 (on LG6). The quantitative trait loci associated with characters detected by composite interval mapping-were not co-localised and revealed a genetic independence. The results obtained in this study show that the most important agronomic and quality traits of brown mustard could be bred independently. Correlation between the studied traits is also discussed. PMID- 15340690 TI - Impact of ecological factors on the initial invasion of Bt transgenes into wild populations of birdseed rape (Brassica rapa). AB - The inevitable escape of transgenic pollen from cultivated fields will lead to the emergence of transgenic crop-wild plant hybrids in natural patches of wild plants. The fate of these hybrids and that of the transgene depend on their ability to compete with their wild relatives. Here we study ecological factors that may enhance the fitness of genetically modified hybrids relative to wild plants for a Bacillus thuringiensis ( Bt) transgene conferring resistance to insects. Mixed stands of wild plants and first-generation hybrids were grown under different conditions of herbivore pressure and density, with Bt oilseed rape ( Brassica napus) as the crop and B. rapa as the wild recipient. Biomass and fitness components were measured from plant germination to the germination of their offspring. The frequency of transgenic seedlings in the offspring generation was estimated using the green fluorescent protein marker. The biomass of F(1) Bt-transgenic hybrids relative to that of wild-type plants was found to be sensitive to both plant density and herbivore pressure, but herbivore pressure appeared as the major factor enhancing their relative fitnesses. In the absence of herbivore pressure, Bt hybrids produced 6.2-fold fewer seeds than their wild neighbors, and Bt plant frequency fell from 50% to 16% within a single generation. Under high herbivore pressure, Bt hybrids produced 1.4-fold more seeds, and Bt plant frequency was 42% in the offspring generation. We conclude that high-density patches of highly damaged wild plants are the most vulnerable to Bt-transgene invasion. They should be monitored early to detect potential transgene spread. PMID- 15340691 TI - A large-scale study of rice plants transformed with different T-DNAs provides new insights into locus composition and T-DNA linkage configurations. AB - Transgenic locus composition and T-DNA linkage configuration were assessed in a population of rice plants transformed using the dual-binary vector system pGreen (T-DNA containing the bar and gus genes)/pSoup (T-DNA containing the aphIV and gfp genes). Transgene structure, expression and inheritance were analysed in 62 independently transformed plant lines and in around 4,000 progeny plants. The plant lines exhibited a wide variety of transgenic locus number and composition. The most frequent form of integration was where both T-DNAs integrated at the same locus (56% of loci). When single-type T-DNA integration occurred (44% of loci), pGreen T-DNA was preferentially integrated. In around half of the plant lines (52%), the T-DNAs integrated at two independent loci or more. In these plants, both mixed and single-type T-DNA integration often occurred concurrently at different loci during the transformation process. Non-intact T-DNAs were present in 70-78% of the plant lines causing 14-21% of the loci to contain only the mid to right border part of a T-DNA. In 53-66% of the loci, T-DNA integrated with vector backbone sequences. Comparison of transgene presence and expression in progeny plants showed that segregation of the transgene phenotype was not a reliable indicator of either transgene inheritance or T-DNA linkage, as only 60 80% of the transgenic loci were detected by the expression study. Co-expression (28% of lines) and backbone transfer (53-66% of loci) were generally a greater limitation to the production of marker-free T(1) plants expressing the gene of interest than co-transformation (71% of lines) and unlinked integration (44% of loci). PMID- 15340692 TI - Isolation and structure elucidation of azoricasterol, a new sterol of the deepwater sponge Macandrewia azorica. AB - Chemical investigation of the deepwater sponge Macandrewia azorica, collected from the flanks of the Gettysburg and Ormonde Sea Mount, North Atlantic, from a depth of 600 m, has led to the isolation of a new sterol with an unusual side chain (1), along with S-methylergothioneine (2). The structures of compounds 1 and 2 were established by employing spectroscopic techniques (NMR, MS, UV, IR and polarimetry). This is the first report of metabolites of a sponge belonging to the genus Macandrewia. PMID- 15340693 TI - Genetic variation in natural honeybee populations, Apis mellifera capensis. AB - Genetic variation in honeybee, Apis mellifera, populations can be considerably influenced by breeding and commercial introductions, especially in areas with abundant beekeeping. However, in southern Africa apiculture is based on the capture of wild swarms, and queen rearing is virtually absent. Moreover, the introduction of European subspecies constantly failed in the Cape region. We therefore hypothesize a low human impact on genetic variation in populations of Cape honeybees, Apis mellifera capensis. A novel solution to studying genetic variation in honeybee populations based on thelytokous worker reproduction is applied to test this hypothesis. Environmental effects on metrical morphological characters of the phenotype are separated to obtain a genetic residual component. The genetic residuals are then re-calculated as coefficients of genetic variation. Characters measured included hair length on the abdomen, width and length of wax plate, and three wing angles. The data show for the first time that genetic variation in Cape honeybee populations is independent of beekeeping density and probably reflects naturally occurring processes such as gene flow due to topographic and climatic variation on a microscale. PMID- 15340694 TI - [Right ventricular function in ARDS and mechanical respiration]. AB - The right ventricle is the stepchild of intensive care medicine. In diseases of the lung mainly when the relationship between ventilation and perfusion is disturbed, assisted respiration with positive end-expiratory pressure (PEEP) is essential to improve oxygenation. The serious damage to the lung parenchyma as seen in adult (acute) respiratory distress syndrome (ARDS) and pneumonia has considerable consequences for cardiac function. Whereas left ventricular function remains almost completely unaffected well into late stages of the disease, the right ventricle is subjected early to stress from the underlying disease and mechanical ventilation. The effects of therapeutic measures aimed at maintaining oxygenation and ventilation partially have negative consequences for right ventricular function and encourage the development of acute cor pulmonale. They can be the cause of right-sided heart failure. PMID- 15340695 TI - [Circadian rhythms and clinical pharmacology]. AB - Almost all physiological functions in animal and man including vital signs display significant daily variations. The existence of internal clock(s) triggering circadian rhythms is now well established. In man, also the onset of certain diseases such as asthma attacks, coronary infarction, angina pectoris and peptic ulcers is not randomly distributed over 24 hours of a day. These rhythmic changes may have implications for drug therapy: In man more than 60 different drugs were shown to exhibit pronounced daily variations in their pharmacokinetics and/or in their effects or side effects. This data demonstrate that the time of day has to be taken into account as an additional parameter influencing the pharmacokinetics, the efficacy and the therapeutic range of drug therapy. PMID- 15340696 TI - [Relevance of sleep for patients with lung diseases]. AB - Sleep is characterized by a profound change of load and capacity of the respiratory system. Load increases due to a rise in upper and lower airway resistance. Capacity decreases due to reduced chemosensitivity, a decrease in muscle activity and minute ventilation. Whereas these changes do not lead to relevant blood gas changes and do not disturb sleep in healthy subjects, patients with respiratory diseases frequently show the first symptoms of their disease during sleep. Pulmonary diseases in which sleep plays an important role are asthma, COPD, hypercapnic respiratory failure, sleep disordered breathing, the overlap-syndrome and cystic fibrosis. Medical history should include sleep and complaints during the night. In asthmatics peak-flow measurements during the night may provide valuable information. In all other disorders mentioned, nocturnal ambulatory recording of respiration and arterial oxygen saturation often allow the detection of relevant disorders of breathing during sleep. If ambulatory monitoring reveals relevant pathology, then further evaluation and treatment in the sleep laboratory are warranted. PMID- 15340697 TI - [A 77-year-old patient with suspected left atrial tumor]. AB - A 77-year-old female presented with a suspected cardiac tumor and thrombosis of both legs diagnosed by phlebography with pulmonary embolism of both sides. In transthoracic echocardiography, transesophageal echocardiography, and CT no intracardial tumor was seen. There was a hiatal hernia which compressed the left atrium from outside. As echocardiographic criteria of hiatal hernia described by D'Cruz we found the left atrium filled with a solid mass and variation of compression of the left atrium depending on breathing. Because of the low incidence of cardiac tumors, differential diagnosis should be done if a cardiac tumor is suspected. A hiatal hernia should always be taken into consideration. PMID- 15340698 TI - [Recommendations for statin therapy in the elderly]. AB - Elderly patients are significantly less likely to receive statins than younger patients possibly because of doubts regarding compliance or concerns regarding the increased likelihood of adverse events and drug interactions. Poor compliance can be expected especially in patients suffering from dementia or depression as well as those whose stage of cardiovascular disease exhibits few symptoms. On the other hand, the clinical significance of CHD events is high in the elderly, and 80% of coronary deaths occur in patients aged over 65 years. The average statistical life expectancy of elderly and old patients is often underestimated. The HPS and PROSPER studies showed that statins reduce mortality and morbidity even in very elderly individuals with a high global cardiovascular risk and/or CAD. Patients up to the age of 79 years should be treated according to the same guidelines as younger patients. Statin therapy should only be considered for patients aged 80 years and older who are at a very high risk for cardiovascular events. PMID- 15340701 TI - [Hydrodissection for tonsillectomy. Results of a pilot study--intraoperative blood loss, postoperative pain symptoms and risk of secondary hemorrhage]. AB - BACKGROUND: : Tonsillectomy is one of the most common procedures in head and neck surgery. Various techniques have been developed to reduce the risk of postoperative bleeding and postoperative pain. Water-jet technology has been designed for a gentle dissection of parenchymal organs. We report the results of a first pilot study using the water-jet technique for tonsillectomy.METHODS.: Water-jet-cutting tonsillectomy was compared with conventional blunt dissection tonsillectomy. 60 Patients were stratified in two groups within a preliminary prospective randomised study. Postoperative pain scores, frequency of postoperative bleeding and intraoperative blood loss were documented.RESULTS.: There was no difference in the average duration of the surgical procedure. The intraoperative bloodloss was markedly decreased in water-jet-cutting tonsillectomy. Postoperative bleeding and postoperative pain were decreased in the water-jet-cutting group.CONCLUSION.: The results of this pilot study demonstrate that water-cutting tonsillectomy is associated with less intraoperative bleeding, reduced frequency of postoperative bleeding events and with less postoperative pain. PMID- 15340702 TI - [Diagnostics of diseases and the function of the facial nerve]. AB - The facial nerve has a complicated course from the brainstem to the periphery. It not only contains motor, but also secretory, sensory, and sensitive fibres. Thus, the functional measure of symptoms can be multi-faceted. The nerve is not directly accessible over an extensive distance because of its long course through the temporal bone. Therefore, diagnostics of nerve function and the differential diagnostics of its diseases could be a great challenge for the otolaryngologist. In this review, the most important methods for clinical examination, electrodiagnostics, and modern imaging techniques are critically surveyed. In addition, the significance of facial nerve monitoring for surgery in the cerebello-pontine angle, parotid surgery, and ear surgery is presented. PMID- 15340703 TI - [Gradenigo syndrome--still a threatening complication of otitis media]. AB - Intracranial complications of acute otitis media are rare in the time of a specific antibiotic therapy. We report about the case of a 13-year-old female patient with an incompletely cured otitis media which was followed by a petrous apicitis presenting abducens nerve palsy. She was successfully treated by a combination of surgical intervention, high dose intravenous antibiotics, systemic corticoid-therapy and hyperbaric oxygen therapy. PMID- 15340704 TI - [Multiple skin-colored moles on the vulva]. PMID- 15340705 TI - [Progressive symmetric erythrokeratodermia of Darier-Gottron]. AB - A 5 1/2-year-old boy revealed symmetric erythematous plaques on both arms and legs as well as in the face. Additionally, contractions of several digital joints were noted. We diagnosed a progressive symmetric erythrokeratoderma and initiated a topical therapy with tretinoin. Here we discuss the etiology, differential diagnoses, and therapeutic options of this rare disorder of keratinization. PMID- 15340706 TI - [Incontinentia pigmenti in a five-week-old girl]. AB - Shortly after birth, a five-week-old female infant developed small blisters and erythema that followed the lines of Blaschko on the upper and lower extremities as well as the abdomen. Histological examination confirmed the clinical presumptive diagnosis of incontinentia pigmenti. We discuss the dinical features, diagnosis, and the molecular genetic basis of this rare inherited skin disorder. PMID- 15340707 TI - [Treatment-resistant granulomatous rosacea-like dermatitis in a 9-year-old girl]. AB - A 9-year-old female developed facial papules and pustules since four years. Clinically, perioral dermatitis was suspected. Different topical therapy regimens and systemic anibiotics had been unsuccessful and a skin biopsy showed granulomatous (lupoid) rosacea. Only systemic antibiotic treatment with minocyclin led to healing of the skin lesions. While granulomatous rosacea-like dermatitis is more frequently diagnosed in adults, it is only rarely encountered in children where, in most of the cases, it represents a therapeutic challenge. PMID- 15340708 TI - [Identification of geophilic and zoophilic dermatophytes in siblings with tinea capitis. A pathogenic factor or contamination?]. AB - Two siblings of African origin presented with multiple scaling patches and alopecia on the scalp four weeks after returning from a vacation in Eritrea. Direct KOH examination revealed fungal elements; Trichophyton mentagrophytes and Trichophyton terrestre were identified in the fungal culture. We discuss the putative pathogenic role of both microorganisms in causing disease. Although infection with Microsporum canis currently accounts for almost fifty percent of all cases of tinea capitis in Germany, other fungi have gained importance due to tourism and increasing migration. PMID- 15340709 TI - [Inherited metabolic disorders with cutaneous manifestations]. AB - Over the last years, the genetic basis of several monogenic inherited metabolic diseases has been elucidated. Interestingly, some of these disorders manifest with characteristic cutaneous symptoms that are often crucial for diagnosis. In most cases, however, besides the skin other organs are affected. Therefore, an interdisciplinary supervision of these patients is highly important. In this review we will discuss diseases that constitute a challenge not only for dermatologists but also for physicians from other specialties. A particular emphasis is put on genetic and clinical features of these disorders as well as current therapeutic concepts. PMID- 15340710 TI - [Hereditary blistering disorders]. AB - Epidermolysis bullosa (EB) is a group of genetic skin disorders whose common feature is the formation of blisters following minor trauma. They present with a wide clinical spectrum of manifestations because of a variety of molecular defects. In patients with mild phenotypes, only skin is affected. The most severe EB forms are multiorgan disorders with a poor prognosis. EB arises from abnormalities in proteins of the dermal-epidermal junction. These specialized protein components aggregate to form anchoring complexes, which attach the epidermis to the dermis. Three major EB-forms can be distinguished on the basis of ultrastructural blistering level: EB simplex--epidermolytic, junctional EB--in the lamina lucida and dystrophic EB--dermolytic. To establish a provisional diagnosis for an EB patient, clinical data, family history and morphologic examination of the skin, e.g. by antigen-mapping, are needed. Complete knowledge of the genetic defect provides the basis to a rational genetic counseling and prenatal testing. Treatment of EB is based on wound care; multidisciplinary management of cases with severe course is required. PMID- 15340711 TI - [Pruritus in childhood. A diagnostic and therapeutic challenge]. AB - From the clinician's point of view, pruritus in children is quite frequent. It mainly occurs along with dermatoses but rarely with systemic diseases such as renal and liver failure or with genetic disorders. Mostly, it appears in the setting of atopic dermatitis (AD). Other frequent differential diagnoses comprise e.g. scabies, impetigo, varicella, tinea, urticaria, mastocytosis and psoriasis. In children, pruritus is most often associated with severe scratching leading to artefacts. This group of patients requires a therapeutical regimen of its own. The use of topical and systemic treatments depends on the underlying aetiology of pruritus and the stage and status of the skin. The physician has to consider that topically applied drugs may cause intoxication due to the different body volume/body surface proportion, especially in newborns and infants. The dosages of systemic drugs need to be adapted in children and UV phototherapy should be performed with caution due to possible longterm photo damage of the skin. Physicians feel more insecurity treating pruritus in children, especially when systemic treatments are taken into consideration. We want to highlight the major aetiologies of pruritus in children and point out the cornerstones of antipruritic therapy in this challenging group of patients in recognition of our own clinical experiences and the current literature. PMID- 15340712 TI - [54th annual meeting of the DSTDG in Marburg]. PMID- 15340713 TI - [To err is human--social and medical aspects of mental illnesses]. PMID- 15340714 TI - [On the cultural history of psychiatry]. AB - About 1800, psychiatry was established as a medical discipline with special institutions (madhouses). Therefore, historiography of psychiatry focuses generally on the last 200 years. This contribution will also illustrate aspects of medical and cultural history, which nowadays are mostly supposed to be less important: the premodern concept of melancholy and hypochondria between humoral pathology, demonology, and psychology; the assessment of psychiatric illness as a "creative malady," even complementary to genius; the dialectics of psychiatric therapies between suppression and emancipation, which is especially prominent in the early nineteenth century in regard to "moral treatment" ( psychische Kur in German), a topic stressed vigorously by the "antipsychiatry" movement around 1970; the denunciation of patients and sections of the population by eugenics ( Rassenhygiene in German) and racism (especially toward the Jews) by psychiatrists. Finally, the miraculous mechanisms of mass hysteria of "normal" individuals are questioned. PMID- 15340715 TI - [Depressive disorders and conduct disorders in childhood and adolescence. Prevalence, course, and risk factors]. AB - Mental disorders belong to the most frequent disorders in the community and lead to noticeable functional impairments. The lifetime prevalence of clinical depression (ICD-10 diagnoses F33, F34) up to age 25 is 12.7%, showing a female male ratio of 2:1. From adolescence onwards, persistence rates of depressive disorders are comparably as high as those found in externalizing disorders. Subclinical depression (ICD-10 subthreshold disorders) at ages 8 and 13 increases the risk for later clinically relevant mental disorders. Conduct disorders (ICD 10 diagnoses F91, F92) are the most frequent mental disorders in children and adolescents with lifetime prevalence rates of 22.4% up to age 25. Conduct disorders show unfavorable courses beginning at preschool age. Precursors of later disorders can be detected as early as toddlerhood. Adverse family factors in childhood and early externalizing problems of the child were most predictive for later conduct disorders. Therefore, the need for early prevention of conduct disorders is highlighted. The focus should be on families with low socioeconomic status (objective: strengthening family and child resources). For depressive disorders, we recommend testing and evaluating the indicated prevention programs in adolescence (objective: strengthening the resources of the adolescent). PMID- 15340716 TI - [Mental disorders in the community: healthcare utilization and disability days]. AB - Mental disorders are considered to be costly in terms of elevated rates of healthcare utilization and increased disability days. For the general German population there has been a lack of data on healthcare utilization and disability rates. The available administrative data are incomplete and problematic because they are based on routine clinical diagnoses that lack reliability. Using the German Health Interview and Examination Survey 1998/99 and its Mental Health Supplement (GHS-MHS) data, this paper examines healthcare utilization and disability associated with mental disorders in a representative sample of the general adult population (18-65 years, n=4181). After reviewing the size and treatment status of mental disorders in Germany, data about primary care or specialist consultations, inpatient treatment, and disability days in the last year are presented for the following groups: (1) subjects who never met the criteria of the diagnosis of a mental disorder, (2) subjects who had one or more mental disorders in the past but none in the past year ("remitted") and (3) subjects who currently (within the last year) met the criteria of one or more mental disorders. We found increased healthcare utilization as well as more disability days in those individuals with a current diagnosis. Remitted persons, however, showed results similar to those who never had a diagnosis of a mental disorder during their lifetimes. The results are consistent with the episodic course of many mental disorders and furthermore might indicate that treatment of mental disorders might contribute to a substantial reduction of direct and indirect illness costs. PMID- 15340717 TI - [Health economic aspects of medical care and therapeutic options for schizophrenic and depressive patients in Germany]. AB - Optimal and effective medical care of patients suffering from psychiatric disorders and their integration into society leads undoubtedly not only to a higher quality of life of the person affected, but also to a reduction of direct and indirect disease-related costs such as loss of earnings and disability pension. Both schizophrenia and depressive disorder display an early age of onset and inclination to a chronic course under inadequate medical care and thus are interesting examples for diseases with enormous direct and indirect disease related health costs. We want to illustrate with these diseases the necessity for further effort, more extensive financial support, and the will for change to maintain the standard of medical care for psychiatric patients which has been achieved during the last 20 years in Germany. To achieve this goal, all sectors of the healthcare system have to recognize the health economic effects of inadequate medical care of psychiatric patients either as a result of understaffing, insufficient application of therapeutic options, inadequate exploitation of the care system, or as an effect of inaccurate legislation and to draw the right conclusions together. Furthermore, more research on the care system of psychiatric patients dealing with economic aspects is required. PMID- 15340718 TI - [Disease management programs in Germany. Current focus and further development]. AB - The structure of the German healthcare system impedes well-coordinated long-term care of the chronically ill. Based on centrally defined standards, German sickness funds have implemented nationwide disease management programs for patients with diabetes. Breast cancer programs are underway. The programs are evidence based. Performance standards are centrally defined and subject to accreditation by a federal office. The programs are designed to improve the coordination of care between the different sectors, but the associated administration costs are often criticized. PMID- 15340719 TI - [Survey among local health departments concerning the implementation of the new infectious disease reporting system]. AB - The Infectious Disease Control Act enacted in Germany in January 2001 led to the establishment of a new reporting system for infectious disease. The implementation of this system was evaluated to identify opportunities for further improvement. In a survey of all German local health departments the following criteria were analyzed: resources (staff and technical equipment), information needs (satisfaction with current offers/further training requirements), data analysis (extent of local data analysis/feedback of national data), and acceptability (case definitions/electronic reporting). In local health departments, 11% of the staff were assigned to the infectious disease reporting system. Data were processed mainly by nonmedical staff (78.4%). A computer work station is available for most staff members. One-third of the local health departments uses the RKI software "SurvNet@rki" for data transmission. All others use commercial software. Experience with the electronic reporting system was rated as very good/good by 47.1% of local health departments, as satisfactory by 44.5%, and as problematic by 8.4%. Most of the local health departments were satisfied with the offers of information provided by RKI (96.4%) and state health departments (83.7%), respectively. However, 49.1% of the local health departments saw a need for further education and training. The implementation of case definitions was supported by 95% of the local health departments, but transmission criteria were criticized. In summary, the new infectious disease reporting system in Germany was successfully implemented. However, the system could be improved through reduction of software problems concerning electronic data processing and transmission, expansion of current offers of in-formation and provision of special staff training programs, and revision of the case definitions concerning clearness and simpler handling. PMID- 15340720 TI - [The new indicator set for health reporting activities in the German States]. AB - In May 2003, the third revised version of the indicator set for health reporting activities was confirmed by the health ministries of all German States (Bundeslander). Modeled on the restructured indicator set which has been annotated with meta-data descriptions, most Bundeslander have now started to collect data for their specific health reporting activities. Thanks to the support provided by national data holders and the Federal Statistical Office, it has been possible to further enlarge the database and for the first time also ensure access via the Federal Statistical Office. In this contribution the authors describe the methodological and statistical principles of the indicator set. Another aspect is the benefit of the indicator set for the health reporting activities in the German States. PMID- 15340721 TI - [Evaluation of the 2003 tobacco tax increase in Germany]. AB - Implementation, acceptance, and effects of the increase in tobacco tax were examined. For this purpose representative samples were assessed before and after the increase of tobacco tax. Furthermore, official data on tax receipts were analyzed. The increase of tobacco tax was passed only partly to the consumer. 57 smokers (4.0%) stopped smoking because of the increase, and 11.9% reduced the amount of cigarettes consumed. The average number of cigarettes consumed was decreased by 0.06% (price elasticity of -0.01). On the basis of the official data, a relative decrease of 5.8% of cigarettes was observed (price elasticity of -1.04 for cigarettes from vending machines). The rather slight increase of price caused smokers to reflect on their smoking behavior. The amount of cigarettes used changed in the official data and in the subsample of younger subjects. PMID- 15340722 TI - [Environment-related health disorders. Experience and perspectives in the care of patients with environment-related health disorders]. AB - Environmental medicine outpatient clinics, counseling centers, and practicing physicians have observed environment-related health disorders in patient groups of mixed age as well as for groups consisting only of adults or children. Practicing physicians suspected correlations between environmental factors and health disorders in 36-45% of cases, environmental medicine outpatient clinics and counseling centers in 4-34% for mixed-age groups, 0-24% for adults, and 9-13% for children. A comparison of these data is difficult due to differences in data acquisition, evaluation methods, and descriptive statistics used. Furthermore, data on children are insufficient. Patient-oriented environmental medicine faces a number of problems regarding determination of exposure, effects, and susceptibility, including a lack of scientifically verified cause-and-effect models as well as incorrect diagnoses, attributions, and conclusions. In view of the scope and intensity of environment-related health disorders, the topic cannot be ignored. A functioning program of environmental medicine counseling and patient care is needed for practicing physicians, universities and/or the public sector to deliver effective primary medical care in this field. As always, the building blocks of environ-mental medicine counseling are medical history, physical examination, differential diagnosis, human biomonitoring, and on-site inspection with environmental monitoring while also taking gender differences into account. Uniform basic documentation procedures and health science analyses will help to optimize patient care in environ-mental medicine. The value of a diagnostic algorithm in the care of patients with environment-related health disorders is beyond dispute. Last but not least, quality assurance and control are a sine qua non of patient-oriented environmental medicine. PMID- 15340723 TI - [Fifty years of nutrition counseling. Remarks and perspectives]. AB - Fifty years of nutrition information and education did not reach their goals. Nutrition-dependent diseases, obesity, and misinformation are still increasing. Cognitive information about nutrition does not induce changes in primary emotion controlled eating habits. Better knowledge of nutrition on the part of consumers mainly activates their bad conscience when they rate their own eating habits. Future prevention campaigns, which clearly address consumer needs using social marketing principles, will be better able to change eating behavior. PMID- 15340724 TI - [11th Retrovirus Conference in San Francisco, 8 to 11 February. 1: Epidemiology, risk and preventive factors, HIV primary and superinfections, mother-child transmission]. PMID- 15340725 TI - [Recommendation of the German Environmental Agency on continuing use of KTW recommendations in the transitional period until EAS becomes official]. PMID- 15340726 TI - [Migration methods and models for estimating possible exposure to textile adjuvants and dyes from clothing textiles during normal use. From the "Public Health Evaluation of Textile Adjuvants and Dyes" Committee of the BfR. "Textiles" Study Group]. PMID- 15340728 TI - ["Topless" cardiopulmonary resuscitation. Fashion or science?]. AB - A decade after the onset of a discussion whether ventilation could be omitted from bystander basic life support (BLS) algorithms, the state of the evidence is reevaluated. Initial animal studies and a prospective randomized patient trial had suggested that omission of ventilation during the first minutes of lay cardiopulmonary resuscitation (CPR) did not impair patient outcomes. More recent studies demonstrate, however, that this may hold true only in very specific scenarios, and that the chest compression-only technique was never superior to standard BLS. Instead of calling basics of BLS training and practice into question, more and better training of lay persons and professionals appears mandatory, and targeted use of dispatcher-guided telephone CPR should be evaluated and, if it improves outcome, it should be encouraged. Future studies should focus much less on the omission but on the optimization of ventilation under the specific conditions of CPR. PMID- 15340729 TI - Diabetes and the heart. PMID- 15340730 TI - Diabetes mellitus and cardiovascular disease. Pandora's box has been opened. AB - As a result of an aging, increasingly obese, and decreasingly physically active population, the global incidence and prevalence of type 2 diabetes mellitus (DM) is increasing. While little can be done to influence the contribution of aging to the increasing global prevalence of diabetes, opportunities abound to develop effective interventions with regard to diet, exercise and obesity to prevent type 2 DM and more importantly, diabetes-associated complications. The major cause of morbidity and mortality from diabetes is from atherosclerotic macrovascular disease-including peripheral, cerebrovascular, and coronary artery disease. The markedly increased risk for cardiovascular disease associated with diabetes compound medical and public health challenges in the face of the burgeoning global epidemic of diabetes and will inevitably strain healthcare resources. Although the cardiovascular benefits of glycemic control have yet to be clearly established, a number of therapeutic interventions improve cardiovascular risk among the high-risk cohort of patients with diabetes. The development of new strategies targeting both primary prevention of diabetes and the prevention of diabetic complications will remain important research and clinical objectives. Great advances are being made in the realm of diabetes and cardiovascular disease, and there is much optimism for the future. PMID- 15340731 TI - [The metabolic syndrome in patients with type 1 diabetes mellitus. Associations with cardiovascular risk factors and cardiovascular morbidity]. AB - BACKGROUND AND PURPOSE: Type 1 diabetes is known to be associated with increased cardiovascular disease in the presence of nephropathy and hypertension. It was the aim of the present study to elucidate whether or not clinical findings of metabolic syndrome (MS) are further increasing cardiovascular morbidity among type 1 diabetics. METHODS: In the present cross-sectional study, 1,241 type 1 diabetics were included. These patients attended the Diabetes Clinic Karlsburg, Germany, from February 1, 2002 to December 31, 2003. The presence of the following findings was taken into consideration as clinical features of MS in type 1 diabetes: fasting triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), body mass index (BMI), daily insulin requirement/kg body weight (b.w.), increased blood pressure > 130/85 mmHg, including overt arterial hypertension. In each of the five categories the highest quintile in each sample was assessed: TG 2.9 +/- 3.6 mmol/l, HDL-C 1.48 +/- 0.46 mmol/l, BMI 29.1 +/- 4.98 kg/m(2) height, insulin requirement 0.71 +/- 0.23 IU/kg b.w., systolic blood pressure 130 +/- 12.3 mmHg. MS was defined as the presence of at least three categories. Among 1,241 type 1 diabetics (651 men, 590 women), 226 patients (129 men, 97 women) fulfilled the criteria of MS. The risk of MS was assessed by multiple regression analysis. Risk variables were: age, diabetes duration, sex, glycated hemoglobin (HbA(1c)), actual smoking, neuropathy, albumin excretion rate (AER), regular alcohol consumption, retinopathy, peripheral vascular disease (PVD), coronary heart disease (CHD), TGs, HDL-C, low-density lipoprotein cholesterol (LDL-C), cholesterol, blood pressure increase, BMI, increased insulin requirement, and foot syndrome. After adjusting for age, the variables were separately included into the mathematical model. The risk of MS was assessed after excluding the variables defining MS. RESULTS: Type 1 diabetics with MS were characterized by higher age (46 vs. 36 years; p < 0.01), and longer diabetes duration (19 vs. 16 years; p < 0.01). The risk of MS was independently associated (odds ratios) with higher age (40-59 years; 4.21; p < 0.01), increased HbA(1c) (1.41; p < 0.01), PVD (2.28; p < 0.01), CHD (2.19; p < 0.01), and the foot syndrome (4.17; p < 0.01). There were no significant associations of MS with type 2 diabetes heredity (first and second degree). CONCLUSION: Patients with type 1 diabetes and the presence of findings of MS are suffering from increased cardiovascular morbidity. The risk of MS increases with the age and HbA(1c). Life style factors such as weight gain and muscular inactivity seem to have an influence on the pathogenesis of MS in type 1 diabetes, thereby increasing cardiovascular morbidity. PMID- 15340732 TI - Fibrinogen and fibrin clot structure in diabetes. AB - Diabetes is associated with an increased risk of developing cardiovascular disease, which is not fully accounted for by the accumulation of classic cardiovascular risk factors in patients. Recent evidence has demonstrated fibrinogen to be a powerful independent risk marker for cardiovascular disease in the general population, and it is also likely to contribute toward the increased atherosclerotic risk in diabetes. The etiology of hyperfibrinogenemia in diabetes is likely to be multifactorial, and at present the mechanisms involved have not been clarified. However, insulin, insulin resistance and inflammation are likely to be involved, especially in type 2 diabetes. The influence of diabetes in determining an individual's atherothrombotic risk is likely to extend beyond that of elevated fibrinogen levels, and may also act via changes in the structure and function of the fibrin clot that forms. Further research is needed to determine the mechanisms underlying these changes, which may lead to development of future interventions to reduce the excessive vascular risk associated with this disease. PMID- 15340733 TI - [Postprandial hyperglycemia as a risk factor for cardiovascular disease. Therapy improves prognosis]. AB - Diabetes is a cardiovascular disease, however, up to two decades ago there was no evidence that hyperglycemia itself is an independent risk factor. However consistent data from recently published prospective studies in subjects with impaired glucose tolerance and patients with early type 2 diabetes prove that postprandial/postchallenge hyperglycemia is an important risk factor for cardiovascular disease. Pathophysiological investigations have shown that excessive postprandial hyperglycemia causes a cascade of proatherogenic abnormalities such as oxidative stress, activation of NFkappaB receptor and impaired NO release of the endothelium. Moreover in the last years intervention studies like DIGAMI and a study in critical ill patients have shown that strict normalization of blood glucose control improves life expectancy in seriously ill patients. There are now three studies: STOP-NIDDM, MERIA and IMT study of the common carotid arteries which impressively demonstrate that control of postprandial hyperglycemia may prevent cardiovascular complications to the same degree as reported for statins and AC-inhibitors. Thus control of the glucose trias-HbA(1c), postprandial and fasting plasma glucoses is essentially practice in patients with cardiovascular disease. PMID- 15340734 TI - [Diabetes mellitus type 2: coronary calcifications as a predictor of coronary artery disease]. AB - BACKGROUND AND PURPOSE: Patients with diabetes mellitus have an increased risk of developing cardiovascular disease. Therefore, coronary artery disease (CAD) is the most common cause of mortality in these patients. The early and reliable diagnosis of coronary atherosclerosis is crucial for an effective treatment. Determination of coronary calcifications with multislice computed tomography offers the possibility to detect coronary calcifications as a sign of early coronary atherosclerosis. The present study examined the possibility to predict CAD in patients with diabetes mellitus by determination of coronary calcifications. PATIENTS AND METHODS: 632 patients (417 men, 215 women, age 54.6 +/- 17.3 years) with diabetes mellitus and suspected CAD were examined. All patients underwent coronary angiography. Coronary stenoses with lumen narrowing > 50% were regarded as significant CAD. Within 3 days coronary multislice computed tomography (Sensation 4, Siemens Medical Solutions, Forchheim, Germany) was performed and coronary calcifications were quantified using the volume score. Sensitivity, specificity, negative and positive predictive value for prediction of CAD were determined for different cutpoints. RESULTS: 440 patients showed a significant CAD, 315 patients underwent coronary transluminal angioplasty and 57 patients coronary artery bypass surgery. Mean volume score was 421 +/- 461. Volume score increased from 2 +/- 6 for patients < 40 years to 751 +/- 801 for patients > 70 years. Women showed a significantly lower score in all age groups. The mean volume score was significantly higher in patients with CAD compared to those without CAD (587 +/- 642 compared to 40 +/- 53; p < 0.01). In all patients without coronary calcifications, CAD could be ruled out angiographically. Using score 0, 10, 100 as cutoff points for the prediction of CAD, a sensitivity of 100%, 97%, and 87% at a specificity of 25%, 69%, and 82% was calculated. Best results were achieved using the 75th percentile as cutoff point with a sensitivity of 91% and a specificity of 84%. In total, CAD could be diagnosed correctly by quantification of coronary calcifications in 94% of all patients. CONCLUSION: Determination of coronary calcifications by multislice computed tomography could be used in patients with diabetes mellitus to detect coronary atherosclerosis and allows the early and exact diagnosis of CAD. PMID- 15340735 TI - [Diabetic nephropathy - current concepts in early diagnosis and treatment of diabetic microvascular complications]. AB - Diabetic nephropathy represents the most important microvascular complication in long-term diabetes mellitus because chronic renal insufficiency is further aggravated by increased cardiovascular morbidity and mortality in diabetic patients. Although early intensive insulin therapy has led to a significant reduction of incidence and prevalence of end-stage renal failure over the last decades in juvenile type 1 diabetes mellitus, the total number of type 2 diabetic patients with chronic renal insufficiency is dramatically increasing due to the improved life expectancy of the general population and the more effective medical treatment of macrovascular complications such as arterial hypertension, coronary artery disease, and peripheral arterial occlusive disease. Apart from the personal burden for each individual the frightening epidemiologic dimension of diabetic nephropathy represents an outstanding challenge for our social systems. PMID- 15340736 TI - [AGE-RAGE: a hypothesis or a mechanism?]. AB - Acausal relation between hyperglycemia and accelerated atherosclerosis has been recently suggested. The AGE-RAGE interaction is a potential mechanism underlying the accelerated atherosclerosis. Hyperglycemia causes via nonenzymatic glycation the formation of AGEs (advanced glycation endproducts). AGEs as well as other ligands like S100/Calgranulin and Amphoterin mediate receptor-independent and dependent (via the interaction with RAGE) effects. The ligand-RAGE-interaction results in an activation of NF-kappaB, increased expression of cytokines, chemokines, and adhesion molecules and induces oxidative stress. A relevant role of the ligand-RAGE-interaction has been demonstrated in in vivo studies, both for the accelerated atherosclerosis and increased neointima formation in diabetes mellitus. Recent data analysing atherosclerotic lesions of diabetic patients provide further evidence for the pathogenetic role of the RAGE-ligand interaction. In addition, new experimental data established that AGEs interact with other receptors than RAGE, while RAGE interacts with a diverse group of ligands. Thus, further studies are needed for the characterization of the ligand RAGE-interaction. These studies will provide a rationale for the development of new therapeutic approaches for accelerated atherosclerosis in diabetes mellitus. PMID- 15340737 TI - [Cardiovascular effects of oral hypoglycemie drugs]. AB - In the recent years there has been increasing interest in the effects of oral hypoglycemic drugs on the cardiovascular system. This has arisen because of recognitions that thiazolidine-diones, peroxisome proliferators-activated receptor gamma (PPAR-gamma), may have antiatherogenic actions and that sulphonylureas are capable of closing the ATP-dependent potassium channel. PPAR gamma agonists exert antiatherogenic action by inhibition the production of monocyte inflammatory cytokines, inhibition of expression of adhesion molecules in endothelial cells, inhibition of the proliferation of vascular smooth muscle cells and have antioxidative effects. The United Kingdom Prospective Diabetes Study (UKPDS), published in 1998, found that the use of sulphonylureas had no increase in cardiovascular mortality and that metformin therapy in obese individuals with type 2 diabetes mellitus was associated with reduced cardiovascular death. Recently, the STOP-NIDDM trial has been shown that patients with impaired glucose tolerance treated with the alpha-glucosidase inhibitor acarbose had a significant reduction in the risk of cardiovascular disease. Currently, the results of the UKPDS trial are the only available clinical data on which to base the choice of treatment for type 2 diabetic patients. When a glucose-lowering oral drug is considered necessary and is not contraindicated, the firstline choice is a sulphonylurea or a glinide (repaglinide or nateglinide) for diabetics who are not overweight and metformin for those who are. PMID- 15340738 TI - [Insulin therapy improves cardiac autonomic function in type 2 diabetic patients]. AB - BACKGROUND AND AIM: There is scintigraphic evidence that insulin improves cardiac autonomic innervation in diabetic patients. The aim of the study was to assess the effects of insulin therapy on parameters of cardiac reflex tests in type 2 diabetic patients. PATIENTS AND METHODS: 30 type 2 diabetic patients with an HbA(1c) of > 7.5% under oral antidiabetic agents were included into the 4-month follow-up. Insulin therapy was performed according to standardized clinical guidelines. Parameters of five cardiac reflex tests were assessed to study cardiac autonomic function: Coefficient of variation (CV) of heart rate variation (HRV) at rest and during deep breathing, HRV in response to standing (max./min. 30:15-ratio), Valsalva ratio, systolic blood pressure response (> 1 test abnormal = cardiac autonomic neuropathy [CAN]). QT(QTc)-interval was also assessed in the patients. RESULTS: At 4-months follow-up, mean insulin dosage in the patients was 25 +/- 8 IU/day. HbA(1c) was decreased from 9,2 +/- 0,7 to 7.2 +/- 0.6 % (p < 0.001). Three parameters of cardiac autonomic function were significantly improved: CV of HRV at rest 2.79 +/- 0.84 versus 2.96 +/- 0.85 (p < 0,001), CV of HRV during deep breathing 4.15 +/- 1.46 versus 4.38 +/- 1.68 (p = 0.002), 30:15 ratio 1.06 +/- 0.05 versus 1.09 +/- 0.04 (p < 0.05). Length of QTc- Interval changed significantly from 417 +/- 12 ms at month 0 to 398 +/- 11 ms at month 4 (p<0.05). CONCLUSIONS: The study demonstrates that insulin therapy with regular insulin optimizes cardiac autonomic function in type 2 diabetes mellitus. It emphasizes the value of cardiac reflex tests with regard to follow-up of cardiac autonomic function in diabetes. PMID- 15340739 TI - [Inflammation and atherogenesis in diabetes mellitus - new therapeutic approaches]. AB - Atherogenesis is characterized by a chronic process, which commences years before diabetes mellitus becomes overt. Each step of the pathogenetic cascade is accompanied by inflammation. The degree of inflammation is a key determinant for the process of atherogenesis. C-reactive protein is elevated by inflammatory processes, correlates with the atherosclerotic prognosis and is therefore an important marker of inflammation. It denotes early cardiovascular damages and is elevated in acute coronary syndromes. Cardiovascular risk is excessively elevated in diabetic patients and inflammatory mechanisms are predominant in this group. Preventive interventions and therapeutic strategies based on pathophysiological processes are necessary. The importance of treatment of inflammation increases. Glitazones have been demonstrated to act via antiinflammatory effects. They do not only improve glucose utilisation, but also have pleiotropic effects, which target the increased cardiovascular risks in diabetic patients. PMID- 15340740 TI - Management of acute coronary syndrome in diabetes mellitus. AB - Diabetes mellitus is an increasingly prevalent condition in the modern era. Coronary artery disease is the major cause of mortality in this population. Despite advances in the therapeutic modalities available for management of coronary artery disease, diabetic patients tend to have worse short and long term outcomes. Acute coronary syndrome in this patient population should be managed aggressively with antithrombotic and antiplatelet agents, as well as early mechanical reperfusion strategies. Diabetic patients presenting with acute coronary syndrome benefit from intensive glycemic control in the first 48 hours. Percutaneous coronary angioplasty using drug-eluting stents with concomitant glycoprotein IIb/IIIa inhibitors should be considered early in diabetic patients with single vessel and multivessel disease. Long-term preventive measures include aspirin, clopidogrel, statins and angiotensin- converting enzyme inhibitors. PMID- 15340741 TI - Diabetes and percutaneous coronary revascularization in the drug-eluting stent era. AB - Diabetes mellitus has reached epidemic proportions and is associated with decreased event-free survival following coronary revascularization. Although the historical complication rates for diabetic patients following percutaneous coronary intervention have been less than acceptable, the emerging drug-eluting stent technology when coupled with an aggressive adjunctive pharmacological regimen will improve the complication rates following percutaneous revascularization for this high-risk group of patients. This review will focus on the historical data associated with revascularization, percutaneous and surgical, and diabetes mellitus and will highlight the emerging data of drug-eluting stents and adjunctive pharmacology. PMID- 15340742 TI - [Surgical revascularization in patients with diabetes mellitus]. AB - Patients with coronary artery disease (CAD) and diabetes mellitus have an increased risk of mortality when undergoing either interventional or surgical revascularization. However, the rate of necessary reinterventions is significantly lower after surgical revascularization than after percutaneous transluminal coronary angioplasty (PTCA). As yet, no results of long-term follow up after stent implantation are available. The risk for a patient with diabetes mellitus and CAD of dying of myocardial infarction after a bypass operation is significantly lower than after PTCA. Bypass operation with sternotomy in patients with diabetes mellitus carries, however, an increased risk of postoperative mediastinitis, especially when both internal thoracic arteries are used for "totally arterial" revascularization. For this reason the internal thoracic artery should be used only unilaterally in surgical revascularization in these patients. Preoperative and postoperative stabilization of the blood sugar level is very important. Sclerosis of the vessels in close proximity to the heart (ascending aorta, carotid arteries) must be clarified preoperatively. The operation should be carried out particularly carefully, with the wound area kept as small as possible. Reexploration should definitely be avoided. If these guidelines are followed, surgical revascularization in patients with CAD and diabetes mellitus can achieve very good results. PMID- 15340743 TI - [The impact of diabetes mellitus on the results of coronary artery bypass grafting with respect to left ventricular function]. AB - BACKGROUND: Diabetes mellitus is not only an independent risk factor for the development of arteriosclerosis, but also a risk factor for the surgical and interventional treatment of coronary artery disease (CAD). PATIENTS AND METHODS: In 2003, a consecutive series of 2,142 patients underwent isolated coronary bypass grafting at the authors' institution, 567 of these suffering from diabetes mellitus. RESULTS: An analysis of the diabetic and nondiabetic patients revealed a more pronounced risk profile, a significantly reduced left ventricular function and a significantly poorer quality of the coronary arteries in the diabetic group. Perioperative mortality in both groups was not different. The incidence of wound infections, renal failure and neurologic complications was much higher in diabetic patients. CONCLUSION: From these findings it can be concluded that coronary artery bypass grafting in the current era is not associated with a higher perioperative mortality in diabetic patients despite their risk profile at baseline. The risk of wound infections and perioperative renal and neurologic complications is much higher in diabetic patients. PMID- 15340744 TI - Image of the month. Non-invasive coronary angiography in a patient with chronic type-a aortic dissection. PMID- 15340745 TI - [Davos lecture 2004: Pericarditis constrictiva with severe pericardial effusion diagnosed with significant therapeutic consequences]. PMID- 15340746 TI - Cloning and characterization of antikeratin human antibodies using a semisynthetic phage antibody library. AB - Antikeratin autoantibodies (AK auto Abs) are very important elements of the human immune system. To improve the outcome of studies on AK auto Abs, it is necessary to generate antikeratin human monoclonal antibodies. The purpose of present study was to isolate antikeratin human monoclonal antibodies by panning a phage antibody library. A semisynthetic phage antibody library with capacity of 4.0x10(8) members was previously constructed. Panning of the library was performed against human epidermal keratin extracted from psoriatic scales. At the last round of the panning, individual colonies were grown in culture for expression of phage antibodies. Their binding activities and specificities to keratin were determined by ELISA, and positive clones were analyzed by DNA fingerprinting. The selected clones were induced with IPTG to express soluble Fab fragments, which were further characterized by ELISA, immunohistochemistry and Western blotting. Finally, DNA sequencing of the variable regions was performed. A human antibody clone which was able to express soluble Fab fragments and recognize Mr 46,000 keratin (K17) was isolated. DNA sequencing demonstrated that the VH and VL of the antibody came from the human VH1 and Vkappa2 families, respectively. We conclude that phage antibody library technology is a powerful way to generate human monoclonal antibodies. The antikeratin antibody we isolated in the present study would be useful in the research on AK auto Abs. PMID- 15340747 TI - A case of familial tumoral calcinosis in a neonate and review of the literature. AB - BACKGROUND: Tumoral calcinosis occurs in two distinct clinical forms. The sporadic form is secondary to chronic renal failure, hyperparathyroidism, milky alkali syndrome, hypervitaminosis D and other systemic disorders. The familial form is extremely rare (around 100 cases worldwide) and affects patients in the first or second decade of life. It is believed to be transmitted in a dominant autosomal manner with variable clinical expressivity and is manifested as hyperphosphatemia, elevated serum 1,25-dihydroxyvitamin D with juxta-articular tumorous calcifications. Moreover, the theory of a unique dental malformation serving as a screening marker for clinically non-apparent affected individuals is revisited and reconfirmed. CASE REPORT: We present a case of a Caucasian male newborn, as well as a review of the literature with differential diagnostic considerations and their therapeutical implications. PMID- 15340748 TI - Hybrid external fixation of distal tibial fractures: new strategy to place pins and wires without penetrating the anterior compartment. AB - INTRODUCTION: Impalement of the anterior compartment musculature remains a problem in the hybrid external fixation of distal tibial fractures. The purposes of this study were to develop a tensioned wire configuration which does not violate the anterior compartment and to analyze the biomechanical implications of new wire configuration. MATERIALS AND METHODS: Thirty-seven adult volunteers without known pathology around either tibia were recruited. Axial computed tomography of the distal tibia was performed at 5-mm slices from the plafond to the upper margin of the syndesmosis. The wire convergence angle was measured at the 1-, 2-, and 2.5-cm levels using the following landmarks: tibialis anterior tendon (TA), tibialis posterior tendon (TP), peroneus brevis tendon (PB), anterolateral border of the lateral malleolus (LM). Two straight lines were drawn by connecting TA and PB and connecting TP and LM. The wire convergence angle was defined as an acute angle between these two lines. Then the orientation of the bisector axis of the wires was measured. As a second part of this study, a validated three-dimensional hybrid external fixator model was developed using finite elements modeling to analyze the stiffness of the frames constructed according to the measured wire convergence angle and orientation. Five simulated configurations were tested. The stiffness of each frame was analyzed under four load conditions: torsion, axial compression, side bending, and anteroposterior bending. RESULTS: The mean convergence angle was 30 degrees irrespective of the level. The bisector axis was oriented towards the anterolateral direction about 20 degrees from the coronal plane. The stiffness of the frame constructed with a wire convergence angle of 30 degrees and an anterolateral wire orientation of 20 degrees was 20-30% less than that of the frame constructed with 60 degrees wires oriented in a coronal plane. The addition of an anteromedial half-pin increased the stiffness significantly. CONCLUSIONS: Two tensioned wires may be placed without violating the anterior compartment by using the above four clinically identifiable landmarks. Addition of a half-pin on the anteromedial surface of the distal articular fragment makes the frame markedly stiffer than is possible using the standard wire configuration. PMID- 15340749 TI - Acetabular reduction osteotomy using surgical dislocation of the hip joint for treatment of a malunited acetabular fracture. AB - INTRODUCTION: Acetabular fractures remain a challenge for the orthopedic and trauma surgeon, with frequently poor outcome in terms of pain and lack of motion and high rate of posttraumatic arthritis especially in badly reconstructed fractures where the anatomy was not restored. Surgical treatment of malunited acetabular fractures is often necessary, although it can be very complex. CASE PRESENTATION: We report a young woman who sustained both column fracture with central dislocation of the femoral head in which the posterior wall fragment was initially not fixed anatomically. CONCLUSIONS: Surgical dislocation of malunited acetabular fractures is a relatively new therapeutic option that provides full access to the femoral head and acetabulum without compromising the blood supply to the femoral head. Our results show that it can also be of great help in restoring malunited acetabular fractures. PMID- 15340750 TI - Intra-articular synovial hemangioma; a rare cause of knee pain and swelling. AB - INTRODUCTION: Synovial hemangioma is a rare intra-articular benign tumor that is difficult to diagnose because of its nonspecific symptoms. Patients are generally children or young adults with a nontraumatic recurrent swollen painful knee. CASE PRESENTATION: A 24-year-old woman presented with a 20-year history of recurrent pain and swelling in the right knee. Physical examination revealed a small painful point adjacent to the medial side of the patella. Magnetic resonance imaging revealed an intra-articular tumor. After arthroscopic examination, surgical excision was performed. The histology was characteristic of synovial hemangioma. RESULTS: Open complete synovectomy of the circumscribed capillary hemangioma was successful, and the patient remains asymptomatic 2 years after surgery. PMID- 15340751 TI - Stimulatory effect of inulin on intestinal absorption of calcium and magnesium in rats is modulated by dietary calcium intakes short- and long-term balance studies. AB - Previous studies have shown that short-term intake of fermentable oligosaccharides (OS), including inulin, can increase mineral intestinal absorption in humans and animals. While the stimulatory effect of these substances on intestinal magnesium (Mg) absorption is generally high and consistent, their effect on calcium (Ca) seems to depend on experimental conditions, particularly the duration of fermentable OS intake. The aim of this study was to determine how the short- and long-term dietary Ca intake may modulate the effect of inulin on Ca absorption. Sixty male Wistar rats, weighing 275 g, were randomized into two groups to receive or not 10% of inulin in their diet. Each group was divided into three sub-groups to receive one of the following dietary Ca levels 0.25%, 0.50% and 0.75% in their food. The animals were fed fresh food and water ad libitum for 40 days. Apparent intestinal absorptions of Ca and Mg were determined at D13 and D36 of the experiment. As expected, inulin feeding increased Ca and Mg absorption in both periods at all dietary Ca levels. However, the effect of inulin on intestinal Ca absorption was dependent on dietary Ca levels and on experiment duration. In the short-term period, the inulin effect was prominent in the groups receiving high or low Ca levels, but in long-term period inulin improved intestinal Ca absorption much more in the group receiving the low Ca level. In addition, efficiency of intestinal absorption of Ca and Mg (%) was negatively affected by Ca intake levels. These results show that the beneficial effect of inulin on intestinal Ca absorption may be more marked in cases where the Ca intake is low or where the organism's Ca requirement is high. Further studies are required to confirm these results in humans. PMID- 15340752 TI - The antioxidant paradox or antioxidant damage. PMID- 15340753 TI - Sequence analysis and bioinformatics analysis of chromosome 17q25 in familial moyamoya disease. AB - OBJECTS: The pathogenesis of moyamoya disease is still unknown. The present study aimed to find out the responsible genes that are located in the 17q25 locus. METHODS: Considering the function, we selected nine genes as candidates from a total of 65 genes identified in the 9-cM region of D17S785-D17S836 in chromosome 17q25, and performed sequence analysis on the DNA samples obtained from a pedigree of familial moyamoya disease, which showed a complete linkage to the region by a haplotype analysis. Also, we attempted to identify candidate genes that have not been known but might be functionally relevant to the disease among a total of 2,100 expressed sequence tag (EST) sequences using bioinformatics techniques. RESULTS AND CONCLUSION: The sequence analysis could detect no mutation in the nine genes. Nor could we identify a novel candidate gene by the EST analysis. Further studies using alternative approaches are warranted to clarify the pathogenesis of moyamoya disease. PMID- 15340754 TI - Integration of autonomic and local mechanisms in regulating cardiovascular responses to heating and cooling in a reptile (Crocodylus porosus). AB - Reptiles change heart rate and blood flow patterns in response to heating and cooling, thereby decreasing the behavioural cost of thermoregulation. We tested the hypothesis that locally produced vasoactive substances, nitric oxide and prostaglandins, mediate the cardiovascular response of reptiles to heat. Heart rate and blood pressure were measured in eight crocodiles (Crocodylus porosus) during heating and cooling and while sequentially inhibiting nitric-oxide synthase and cyclooxygenase enzymes. Heart rate and blood pressure were significantly higher during heating than during cooling in all treatments. Power spectral density of heart rate and blood pressure increased significantly during heating and cooling compared to the preceding period of thermal equilibrium. Spectral density of heart rate in the high frequency band (0.19-0.70 Hz) was significantly greater during cooling in the saline treatment compared to when nitric-oxide synthase and cyclooxygenase enzymes were inhibited. Cross spectral analysis showed that changes in blood pressure preceded heart rate changes at low frequencies (< 0.1 Hz) only. We conclude that the autonomic nervous system controls heart rate independently from blood pressure at higher frequencies while blood pressure changes determine heart rate at lower frequencies. Nitric oxide and prostaglandins do not control the characteristic heart rate hysteresis response to heat in C. porosus, although nitric oxide was important in buffering blood pressure against changes in heart rate during cooling, and inhibition caused a compensatory decrease in parasympathetic stimulation of the heart. PMID- 15340755 TI - [Do antisense oligonucleotides improve viral immunopathology?]. AB - BACKGROUND: Tumor necrosis factor (TNF)-alpha as a proinflammatory cytokine is of great importance during the development of herpes simplex virus-1 keratitis (HSK). In this study the local administration of antisense oligonucleotides (ASON) targeting TNF-alpha was examined for its usefulness in ameliorating this disease. METHODS: Uptake and efficacy of the oligonucleotides were studied in vitro by fluorescence microscopy and flow cytometry. Substance- and sequence specific influences on the development of HSK were scrutinized in an animal model. RESULTS: Quick and stable uptake of FITC-labeled ASON by isolated spleen and lymph node cells was proved. The production of TNF-alpha by these cells after stimulation with HSV antigen or concanavalin A (ConA) was clearly downregulated after addition of ASON. In vivo, incidence and development of HSK were ameliorated after subepithelial corneal injection of ASON targeting TNF-alpha. When buffer and control oligonucleotides were given, no significant influence on the disease was found. CONCLUSION: The ASON effectively reduced TNF-alpha secretion in vitro and suppressed the development of experimental HSK in vivo. PMID- 15340756 TI - Renal function in the setting of urinary diversion. AB - Incorporating bowel into the urinary tract sets the stage for a potentially dangerous situation for the upper part of this tract. Obstruction, reflux and chronic bacteriuria may develop, all of which can all be detrimental. Most reports on renal function have used IVP and serum creatinine only, methods which are inadequate for proper assessment. Long-term follow-up of patients with ileal conduit diversion reveals a high incidence of morphological and/or functional damage to the kidneys. Refluxing techniques for implanting the ureters have usually been employed. In patients with continent cutaneous diversion or orthotopic bladder substitution, some recent publications have shown rather well preserved glomerular filtration rates. Traditionally, antirefluxing ureteric implantation has been used in these patients. There is presently a trend towards refluxing anastomosis in this setting, providing a low pressure pouch has been constructed. However, pressure can be high in such pouches and bacteriuria is common. The consequences for the fate of the upper urinary tract is unknown and caution should be exercised in recommending such techniques. There is clearly a need for prospective randomized controlled studies on the issue of refluxing versus antirefluxing anastomosis in continent urinary reconstruction. Patients with continent or non-continent diversion should have lifelong follow-up with regard to the upper urinary tract. PMID- 15340757 TI - Bowel function after urinary diversion. AB - Removal of bowel segments for the purposes of urinary diversion may have a significant effect on bowel function. The remaining bowel may not be able to fulfill its normal role, resulting in not only malabsorption syndromes, but also dysfunctional defecation. Provided, however, care is taken in selecting patients and the bowel segments for such procedures and follow-up is vigilant, complications should be minimal. There appears to be a significant risk of developing symptoms of increased bowel frequency following urinary diversion and reconstruction. Patients need to be counselled regarding this prior to surgery, however, further data needs to be collected to accurately quantify the risk and the effect it has on quality of life. PMID- 15340758 TI - Isolation of ESTs from cacao (Theobroma cacao L.) leaves treated with inducers of the defense response. AB - Pathogenic diseases represent a major constraint to the growth and yield of cacao (Theobroma cacao L.). Ongoing research on model plant systems has revealed that defense responses are activated via signaling pathways mediated by endogenous signaling molecules such as salicylic acid, jasmonic acid and ethylene. Activation of plant defenses is associated with changes in the expression of large numbers of genes. To gain a better understanding of defense responses in cacao, we have employed suppressive subtractive hybridization (SSH) cDNA libraries, macroarray hybridization analysis, high throughput DNA sequencing and bioinformatics to identify cacao genes induced by these signaling molecules. Additionally, we investigated gene activation by a phytotoxic elicitor-like protein, Nep1. We have identified a unigene set of 1,256 members, including 330 members representing genes induced during the defense response. PMID- 15340759 TI - Schedule-dependent synergism and antagonism between pemetrexed and paclitaxel in human carcinoma cell lines in vitro. AB - Pemetrexed is a novel multitargeted antifolate with significant clinical activity against a variety of tumors. We studied the schedule-dependent cytotoxic effects of pemetrexed in combination with paclitaxel in vitro to improve our understanding of how this combination might be used clinically. Human lung cancer A549 cells, breast cancer MCF7, ovarian cancer PA1, and colon cancer WiDr cells were exposed to both pemetrexed and paclitaxel in vitro. Cell growth inhibition after 5 days was determined and the effects of drug combinations were analyzed by the isobologram method (Steel and Peckham). Simultaneous exposure to pemetrexed and paclitaxel for 24 h produced antagonistic effects in A549 and PA1 cells, additive/antagonistic effects in MCF7 cells, and additive effects in WiDr cells. Pemetrexed for 24 h followed by paclitaxel for 24 h produced synergistic effects in A549 and MCF7 cells and additive effects in PA1 and WiDr cells, while the reverse sequence produced additive effects in all four cell lines. Cell cycle analysis supported these observations. Our findings suggest that the simultaneous administration of pemetrexed and paclitaxel is suboptimal. The optimal schedule of pemetrexed in combination with paclitaxel is the sequential administration of pemetrexed followed by paclitaxel, and this schedule should be assessed in clinical trials for the treatment of solid tumors. PMID- 15340760 TI - Quality-adjusted survival analysis shows differences in outcome after immunosuppression or bone marrow transplantation in aplastic anemia. AB - Bone marrow transplantation (BMT) and immunosuppression (IS) have improved the prognosis of aplastic anemia; both treatments have specific advantages and drawbacks but similar survival rates. Analysis of additional endpoints may help in treatment decisions. In a single-center study, patients with aplastic anemia treated with IS (n=155) or BMT (n=52) were compared for survival, event-free survival, and quality-adjusted time without symptoms and toxicity (Q-TWiST). Probability of overall and event-free survival at 15 years was similar among both groups (BMT 51+/-15% and 25+/-14%, IS 53+/-10% and 27+/-8%), with more early deaths in the transplant group and more late deaths in the IS group. There were differences in terms of mean duration of seven analyzed health states: time with symptoms from treatment-related toxicity (IS 0.36 years, BMT 0.27), transfusion dependency (IS 0.66 years, BMT 0.1 years), partial remission (IS 3.27 years, BMT 1.42), and secondary clonal disorder (IS 0.68 years, BMT 0.04) was significantly longer for IS compared to BMT (p< or =0.001). Patients treated with BMT spent more time with extensive chronic graft-versus-host disease (GvHD) (IS 0 years, BMT 0.96, p<0.023) and in CR without drugs (IS 1.22 years, BMT 2.43, p=0.056). In conclusion, survival, event-free survival, and Q-TWiST are similar. BMT-treated patients had longer periods free from symptoms, while IS-treated patients needed closer medical care, transfusion support, and medications. PMID- 15340761 TI - Hemolytic uremic syndrome following prolonged gemcitabine therapy: report of four cases from a single institution. AB - Hemolytic uremic syndrome (HUS) has been described following the administration of multiple antineoplastic agents, most notably mitomycin C. More recently, several cases of gemcitabine-induced HUS have been observed with the overall incidence of gemcitabine-induced HUS estimated at 0.015-0.25%. We here report on four patients who developed HUS following gemcitabine therapy at our institution within the last year (incidence 1.4%). All these patients had advanced-stage disease, were heavily pretreated, and received prolonged gemcitabine application, suggesting that in this subgroup of patients HUS may be more frequently encountered than documented so far. PMID- 15340762 TI - Expression of Wilms' tumor gene 1 at different stages of acute myeloid leukemia and analysis of its major splice variants. AB - WT1 is a transcription factor involved in differentiation and proliferation of acute myeloid leukemia (AML) blasts and is expressed in 90% of cases, as determined by nested reverse transcription polymerase chain reaction (RT-PCR). It is proposed to be a key molecule in leukemia promotion. To assess the relevance of WT1 expression, we analyzed blood and bone marrow samples from 58 AML patients (37 at diagnosis, 8 in hematological remission, and 13 at relapse) for the level of WT1 expression, using quantitative real-time RT-PCR. In addition, 21 randomly chosen samples were also analyzed for the quantitative expression of the main WT1 splice variants. As expected, samples from patients at the time of diagnosis or relapse showed significantly higher WT1 expression compared to samples from patients in remission or control samples. No striking difference in expression levels was found between various French-American-British (FAB) subtypes. The level of WT1 expression observed in patients at the time of initial diagnosis was similarly high in patients at relapse. Expression of the four main isoforms (E5+/KTS+, E5-/KTS+, E5+/KTS-, and E5-/KTS-) was found in all samples with significantly higher expression levels of the E5+ variants. Together, these findings support the potential of WT1 as a target for novel treatment approaches in AML. PMID- 15340764 TI - Cellular antitumor immune response to a branched lysine multiple antigenic peptide containing epitopes of a common tumor-specific antigen in a rat glioma model. AB - Human malignant gliomas contain epidermal growth factor receptor (EGFR) gene mutations that encode tumor-associated antigens (TAAs) that can be targeted using immunological techniques. One EGFR mutant gene (EGFRvIII) encodes a protein with an epitope that is not found in normal tissues. A number of studies have focused on this unique epitope as a potential target for tumor vaccines. In the present study, we examined the cellular immune effects of a peptide containing multiple copies of the unique EGFRvIII epitope linked together by way of a lysine bridge. Fischer rats were vaccinated with an EGFRvIII multiple antigenic peptide (MAP). While vaccination produced a humoral immune response, anti-MAP antibody production was not accompanied by expression of the Th2 response cytokine IL-4. In MAP/GM-CSF vaccinated animals, a cellular immune response was detected in association with the appearance of CD4+ and CD8+ T cells at the tumor site. Splenocytes and CD8+ T cells from vaccinated rats produced the Th1 cytokine IFN gamma in vitro in response to stimulation by rat glioma cells expressing EGFRvIII, but not by those expressing wild-type EGFR. MAP vaccine also induced a specific lytic antitumor CTL immune response against F98 glioma cells expressing EGFRvIII, but not against F98 cells expressing either wild-type EGFR or no receptor. The in vivo growth of F98(EGFRvIII) cells was attenuated in vaccinated rats; whereas, growth of F98(EGFR) cells was not. The median survival of vaccinated rats was increased 72% over that of unvaccinated controls challenged with intracerebral F98(EGFRvIII) tumor implants. Therefore, MAP vaccination produced a predominantly cellular antitumor immune response directed against F98 gliomas expressing the EGFRvIII target antigen. The potent immunosuppressive effects of F98 glioma cells mimic the human disease and make this particular tumor model useful for studying immunotherapeutic approaches to malignant gliomas. PMID- 15340765 TI - Cancer Immunotherapy 2004: Mainz, Germany, 6-7 may 2004. PMID- 15340766 TI - EEG correlates of coordinate processing during intermanual transfer. AB - Goal-directed movements require mapping of target information to patterns of muscular activation. While visually acquired information about targets is initially encoded in extrinsic, object-centered coordinates, muscular activation patterns are encoded in intrinsic, body-related coordinates. Intermanual transfer of movements previously learned with one hand is accomplished by the recall of unmodified extrinsic coordinates if the task is performed in original orientation. Intrinsic coordinates are retrieved in case of mirror-reversed orientation. In contrast, learned extrinsic coordinates are modified during the mirror movement and intrinsic coordinates during the originally oriented task. To investigate the neural processes of recall and modification, electroencephalogram (EEG) recording was employed during the performance of a figure drawing task previously trained with the right hand in humans. The figure was reproduced with the right hand (Learned-task) and with the left hand in original (Normal-task) and mirror orientations (Mirror-task). Prior to movement onset, beta-power and alpha- and beta-coherence decreased during the Normal-task as compared with the Learned-task. Negative amplitudes over fronto-central sites during the Normal task exceeded amplitudes manifested during the Learned-task. In comparison to the Learned-task, coherences between fronto-parietal sites increased during the Mirror-task. Results indicate that intrinsic coordinates are processed during the pre-movement period. During the Normal-task, modification of intrinsic coordinates was revealed by cerebral activation. Decreased coherences appeared to reflect suppressed inter-regional information flow associated with utilization of intrinsic coordinates. During the Mirror-task, modification of extrinsic coordinates induced activation of cortical networks. PMID- 15340767 TI - Coupling between place cells and head direction cells during relative translations and rotations of distal landmarks. AB - Hippocampal place cells are selectively active when a rat occupies restricted locations in an environment, and head direction cells fire selectively when the rat's head is pointed in a particular direction in allocentric space. Both place cells and head direction cells are usually coupled, and they are controlled by a complex interaction between external landmarks and idiothetic cues. Most studies have investigated this interaction by rotating the landmarks in the environment. In contrast, a recent study translated the apparatus relative to the landmarks in an environment and found that most place cells maintained the same preferred location on the apparatus regardless of the location of the apparatus in the room. Because head direction cells are insensitive to the rat's location in an environment, the distal landmarks may influence the place field firing locations primarily by controlling the bearing of the head direction cell system. To address this question, ensembles of CA1 place cells and head direction cells of the anterior thalamus were recorded simultaneously, as a rectangular or circular track was moved to different locations in a room with distinct visual landmarks. Most place cells maintained their firing fields relative to the track when the track was translated, and head direction cells maintained the same preferred firing direction. When the distal landmarks were rotated around the track, the firing fields of place cells and the preferred directions of head direction cells rotated with the cues. These results suggest that the precise firing locations of place cells are controlled by an interaction between local and idiothetic cues, and the orientation of the CA1 ensemble representation relative to the distal landmarks may be controlled indirectly by the distal landmarks' influence over the bearing of the head direction cell system. PMID- 15340769 TI - Performance of quantitative analyses by liquid chromatography-electrospray ionisation tandem mass spectrometry: from external calibration to isotopomer based exact matching. AB - Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a versatile coupling system which combines both selectivity and sensitivity and certainty. Hence, it is generally considered as the most reliable technique to quantify chemical compounds in complex matrices. In the present paper, we evaluate the performance of LC-MS/MS methods for the quantification of 3-nitrotyrosine in human urine in order to point out its dependence on the design of the quantification method, and emphasize the role of matrix effects in the performance. We compare external and internal calibrations, isotope dilution and isotopomer-based exact matching. The role of both sample preparation and multiple transitions monitoring is particularly addressed. PMID- 15340770 TI - Assigning environmental variables to observed biological changes. AB - A method for assigning environmental variables to observed biological changes in benthic communities is proposed. The approach requires biological and environmental sampling at the same sites. Additionally, a biological gradient or trend such as a change in observed species or a significant change in their relative abundances is necessary in order to connect the biological observations to the environmental measurements. Whether there is a statistical significant correspondence between the environmental measurements and the biological changes is tested after quantifying the biological changes by using the community disturbance index (CDI). Finally, the environmental variables that are most strongly associated with the biological changes are identified, and it is proposed that these are strong candidates as the pollutants responsible for the biological changes observed. However, this cannot be confirmed using the monitored data only. The approach is tested on data collected in monitoring surveys at the Ekofisk oil field in the North Sea. The results indicate the method is feasible for assigning environmental variables to observed biological changes. PMID- 15340771 TI - Arsenic speciation in hair extracts. AB - Ingested arsenic is known to be not only excreted by urine, but to be stored in sulphydryl-rich tissue like hair, nail or skin. We developed an extraction method for arsenic species from these tissues and studied the stability of different arsenic species during the extraction process. Inorganic and pentavalent methylated arsenic was found to be stable under the extraction conditions, whereas trivalent methylated arsenicals and the thio-analogue of DMA(V) (DMAS) showed reduced stability. The absorption ability of hair for these different species was studied as well. Inorganic arsenic is better absorbed by hair than monomethyl- or dimethyl-arsenicals, whereby the trivalent forms are taken up better than the pentavalent forms. Independent of which methylated arsenical was used for the incubation, the pentavalent form was always the dominant form after extraction. Hair and nail samples from humans suffering from chronic arsenic intoxication contained dominantly inorganic arsenic with small and strongly varying amounts of DMA(V) and MA(V) present. DMAS was only found in some nail sample extracts containing unusually high amounts of DMA(V) and is believed to be formed during the extraction process. PMID- 15340772 TI - Speciation of organotin compounds by capillary electrophoresis: comparison of aqueous and mixed organic-aqueous systems. AB - A capillary electrophoresis method with direct ultraviolet detection was developed for the analysis of organotin species. Despite the fact that direct detection of organotin compounds by ultraviolet absorption is difficult because most organotins possess poor chromophoric properties, the application of low wavelength (lambda = 200 nm) and mixed organic-aqueous media enabled a significant enhancement in sensitivity. A mixed organic-aqueous system (10% methanol/40% acetonitrile/50% H2O) containing acetic acid and tetrabutylammonium perchlorate formed the basis for rapid, efficient and sensitive determinations of organotin cations such as tripropyltin, tributyltin, triphenyltin and diphenyltin. The concentration limits of detection (LOD) for the four organotin compounds were in the range of 0.4-14 microM, comparable to that obtained with the most sensitive indirect UV method reported until now, and took advantage of a stable baseline, a symmetric peak shape and an absence of disturbing system peaks. The relative standard deviations (n = 7) for the relative peak time and peak area were 0.44-0.77 and 4.8-5.8%, respectively. In addition to sensitivity enhancements, the use of organic-aqueous systems instead of pure aqueous media resulted in improved selectivity and efficiency of separations. PMID- 15340773 TI - Increased cardiac workload by closure of the ductus arteriosus leads to hypertrophy and apoptosis rather than to hyperplasia in the late fetal period. AB - It is generally thought that adult mammalian cardiomyocytes compensate for an increased workload by hypertrophy, whereas fetal myocardium grows by cellular proliferation. We analyzed the response of late-fetal rat hearts upon an increased workload imposed by premature constriction of the ductus arteriosus with indomethacin. Initially the fetal heart responds by proliferative growth, as both wet weight and labeling index (bromodeoxyuridine incorporation) of the ventricles increased, whereas neither a change in the fibroblast fraction, ploidy and nucleation in the ventricles is observed. However, this hyperplastic growth is abrogated by a subsequent burst in apoptosis and followed by a hypertrophic response as based on a decrease in DNA and increase in both RNA and protein concentration. This hypertrophic growth was accompanied by a 1.4-fold increase in the volume of the cardiomyocytes. Changes in the molecular phenotype characteristic of pressure-overload hypertrophic growth accompany the process. Thus, the levels of expression of beta-myosin heavy chain and atrial natriuretic factor mRNA increased, of sarcoplasmic/endoplasmic reticulum ATPase (SERCA2) mRNA decreased, and of alpha-myosin heavy chain, phospholamban, and calsequestrin mRNA did not change. In situ hybridization showed that the pattern of mRNA expression changed first in the right ventricular wall and subsequently in the left ventricular free wall as well. It is concluded that pressure-overload imposed on the late-fetal heart induces limited proliferative growth complemented by extensive hypertrophic growth. PMID- 15340774 TI - Effects of the atrial antiarrhythmic drug AVE0118 on cardiac ion channels. AB - Previous studies in pigs and goats have demonstrated that AVE0118 prolongs atrial refractoriness without any effect on the QT-interval. The purpose of the present study was to investigate the effect of the compound on various cardiac ion channels. AVE0118 blocked the pig Kv1.5 and the human Kv1.5 expressed in Xenopus oocytes with IC(50) values of 5.4+/-0.7 microM and 6.2+/-0.4 microM respectively. In Chinese hamster ovary (CHO) cells, AVE0118 decreased the steady-state hKv1.5 current with an IC(50) of 1.1+/-0.2 microM. The hKv4.3/KChIP2.2 current in CHO cells was blocked by AVE0118 by accelerating the apparent time-constant of inactivation ( tau(inact)), and the integral current was inhibited with an IC(50) of 3.4+/-0.5 microM. At 10 microM AVE0118 tau(inact) decreased from 9.3+/-0.6 ms ( n=8, control) to 3.0+/-0.3 ms ( n=8). The K(ACh) current was investigated in isolated pig atrial myocytes by application of 10 microM carbachol. At a clamp potential of -100 mV the I(KACh) was half-maximally blocked by 4.5+/-1.6 microM AVE0118. In the absence of carbachol, AVE0118 had no effect on the inward current recorded at -100 mV. Effects on the I(Kr) current were investigated on HERG channels expressed in CHO cells. AVE0118 blocked this current half-maximally at approximately 10 microM. Comparable results were obtained in isolated guinea pig ventricular myocytes, where half-maximal inhibition of the I(Kr) tail current occurred at a similar concentration of AVE0118. Other ionic currents, like the I(Ks), I(KATP) (recorded in guinea pig ventricular myocytes), and L-type Ca(2+) (recorded in pig atrial myocytes) were blocked by 10 microM AVE0118 by 10+/-3% ( n=6), 28+/-7% ( n=4), and 22+/-13% ( n=5) respectively. In summary, AVE0118 preferentially inhibits the atrial K(+) channels I(Kur), I(to) and I(KACH). This profile may explain the selective prolongation of atrial refractoriness described previously in pigs and goats. PMID- 15340775 TI - Effects of flunarizine on spontaneous synaptic currents in rat neocortex. AB - Flunarizine, a non-selective blocker of voltage-dependent Ca(2+) and Na(+) channels, is clinically effective against several neurological disorders, including epilepsy, migraine, and alternating hemiplegia of childhood. We examined the effects of flunarizine on spontaneous post-synaptic currents in acute brain slices maintained in vitro using patch-clamp electrophysiology. Flunarizine significantly attenuated the amplitude of spontaneous currents in pyramidal neurons from juvenile rat neocortex. Flunarizine had no effect on miniature spontaneous events recorded in the presence of tetrodotoxin, a blocker of voltage-dependent sodium channels. In high (9 mM) extracellular potassium, flunarizine reduced the amplitude and frequency of the spontaneous currents. Additionally, dimethyl sulfoxide, the solvent used in our experiments, reduced the amplitude of spontaneous currents, but only in high extracellular potassium. Our data suggest that the clinical activity of flunarizine may in part be a consequence of reducing spontaneous synaptic currents in the neocortex, especially under conditions of heightened neuronal activity. PMID- 15340776 TI - Glutathione-conjugated toluene diisocyanate causes airway inflammation in sensitised mice. AB - Toluene diisocyanate (TDI) is a highly volatile compound that reacts readily with nucleophilic compounds, sulfhydryl groups in particular. Since the epithelial lining fluid of the airways contains high levels of the sulfhydryl, glutathione (GSH), inhalation of TDI is likely to result in the formation of GS-TDI conjugates. We therefore investigated whether GS-TDI is capable of provoking irritant and/or allergic reactions. Irritant effects of GS-TDI were studied after intratracheal administration of a range of doses of GS-TDI in saline to naive BALB/c mice. GS-TDI caused a dose-dependent increase in neutrophils in the lungs 24 h after instillation. A dose equivalent to 150 microg of TDI or lower had no effect. For provocation of allergic reactions, mice were sensitised by application of 1% TDI onto the skin on days 0 and 1, and challenged intratracheally with a sub-irritant dose of GS-TDI on day 8. GS-TDI did not induce non-specific tracheal hyperreactivity to carbachol 24 and 48 h after challenge in TDI-sensitised mice. However, it increased the numbers of neutrophils in the lungs as compared with the control mice. These findings suggest that GSH conjugation does not diminish the capacity of TDI to elicit irritant-induced inflammation in the lungs of mice at doses above 150 microg of TDI in the conjugate. Moreover, the capacity to induce allergic-specific inflammation was retained at concentrations of GS-TDI being devoid of irritant activity. However, the GS-TDI conjugate failed to induce non-specific tracheal hyperreactivity. This may be the consequence of the deposition of excess of GSH upon local dissociation of the conjugate. PMID- 15340777 TI - Induction of preneoplastic rat liver lesions with an attenuated p53 response by low doses of diethylnitrosamine. AB - Previous reports have documented an attenuated p53 response to DNA-damage in preneoplastic enzyme-altered foci (EAF). Data suggest that this alteration is an adaptation to genotoxic stress induced by carcinogens. Here, we have studied whether the altered p53 response in EAF can be related to acutely apoptotic or cytotoxic doses of the carcinogen diethylnitrosamine (DEN). Eight groups of rats received cumulative doses of 0.25, 0.5, 1.0 and 2.0 mmol DEN/kg, administered weekly for either 10 or 20 weeks. A ninth group received 3.0 mmol/kg for 10 weeks, which gave a supralinear EAF response. Twenty-four hours before sacrifice, all rats were given a challenge dose of DEN in order to induce a p53 response in hepatocytes. The numbers of p53-positive hepatocytes in EAF and in surrounding tissue were analyzed. Unexpectedly, all cumulative doses gave rise to p53 negative EAF and 20-week treatment gave larger EAF with fewer p53-positive hepatocytes than 10-week treatment. It was also observed that at the lowest doses, most EAF developed in midzonal areas. Similar results were obtained with aflatoxin B1. Single high doses of DEN induced p53 accumulation and apoptosis within 24 h, whereas lower doses did not. It is concluded that EAF with an attenuated p53 response can be induced by low doses of genotoxic compounds, not giving rise to acute apoptosis or necrosis. Instead, it is suggested that time is an important determinant for its development at low doses and that a delayed type of apoptosis might be important. PMID- 15340778 TI - Alkaliflexus imshenetskii gen. nov. sp. nov., a new alkaliphilic gliding carbohydrate-fermenting bacterium with propionate formation from a soda lake. AB - Anaerobic saccharolytic bacteria thriving at high pH values were studied in a cellulose-degrading enrichment culture originating from the alkaline lake, Verkhneye Beloye (Central Asia). In situ hybridization of the enrichment culture with 16S rRNA-targeted probes revealed that abundant, long, thin, rod-shaped cells were related to Cytophaga. Bacteria of this type were isolated with cellobiose and five isolates were characterized. Isolates were thin, flexible, gliding rods. They formed a spherical cyst-like structure at one cell end during the late growth phase. The pH range for growth was 7.5-10.2, with an optimum around pH 8.5. Cultures produced a pinkish pigment tentatively identified as a carotenoid. Isolates did not degrade cellulose, indicating that they utilized soluble products formed by so far uncultured hydrolytic cellulose degraders. Besides cellobiose, the isolates utilized other carbohydrates, including xylose, maltose, xylan, starch, and pectin. The main organic fermentation products were propionate, acetate, and succinate. Oxygen, which was not used as electron acceptor, impaired growth. A representative isolate, strain Z-7010, with Marinilabilia salmonicolor as the closest relative, is described as a new genus and species, Alkaliflexus imshenetskii. This is the first cultivated alkaliphilic anaerobic member of the Cytophaga/ Flavobacterium/ Bacteroides phylum. PMID- 15340779 TI - Characterization of a heme oxygenase of Clostridium tetani and its possible role in oxygen tolerance. AB - In order to colonize mammalian wounds, the anaerobic bacterium Clostridium tetani must presumably cope with temporary oxic conditions. Therefore, the recently decoded genome sequence was searched for genes which could confer oxygen tolerance. A few identified systems such as superoxide dismutases and peroxidases are probably responsible for this protection against toxic oxygen species. Another system was detected, a heme oxygenase which could have a role in establishing or maintaining an anoxic microenvironment in the process of wound colonization. The hemT gene encoding the heme oxygenase is expressed in C. tetani, as shown by reverse transcription-PCR. When overexpressed in Escherichia coli, the enzyme converts heme to biliverdin under strict oxic conditions. PMID- 15340780 TI - A gas vesiculate planktonic strain of the purple non-sulfur bacterium Rhodoferax antarcticus isolated from Lake Fryxell, Dry Valleys, Antarctica. AB - A moderately psychrophilic purple non-sulfur bacterium, Rhodoferax antarcticus strain Fryx1, is described. Strain Fryx1 was isolated from the water column under the ice of the permanently frozen Lake Fryxell, Antarctica. Cells of Fryx1 are long thin rods and contain gas vesicles, the first report of such structures in purple non-sulfur bacteria. Gas vesicles are clustered at 2-4 sites per cell. Surprisingly, the 16S rRNA gene sequence of strain Fryx1 is nearly identical to that of Rfx. antarcticus strain AB, a short, vibrio-shaped phototroph isolated from an Antarctic microbial mat. Although showing physiological parallels, strains AB and Fryx1 differ distinctly in their morphology and absorption spectra. DNA-DNA hybridization shows that the genomes of strains AB and Fryx1 are highly related, yet distinct. We conclude that although strains AB and Fryx1 may indeed be the same species, their ecologies are quite different. Unlike strain AB, strain Fryx1 has adapted to a planktonic existence in the nearly freezing water column of Lake Fryxell. PMID- 15340781 TI - Seeing green bacteria in a new light: genomics-enabled studies of the photosynthetic apparatus in green sulfur bacteria and filamentous anoxygenic phototrophic bacteria. AB - Based upon their photosynthetic nature and the presence of a unique light harvesting antenna structure, the chlorosome, the photosynthetic green bacteria are defined as a distinctive group in the Bacteria. However, members of the two taxa that comprise this group, the green sulfur bacteria (Chlorobi) and the filamentous anoxygenic phototrophic bacteria ("Chloroflexales"), are otherwise quite different, both physiologically and phylogenetically. This review summarizes how genome sequence information facilitated studies of the biosynthesis and function of the photosynthetic apparatus and the oxidation of inorganic sulfur compounds in two model organisms that represent these taxa, Chlorobium tepidum and Chloroflexus aurantiacus. The genes involved in bacteriochlorophyll (BChl) c and carotenoid biosynthesis in these two organisms were identified by sequence homology with known BChl a and carotenoid biosynthesis enzymes, gene cluster analysis in Cfx. aurantiacus, and gene inactivation studies in Chl. tepidum. Based on these results, BChl a and BChl c biosynthesis is similar in the two organisms, whereas carotenoid biosynthesis differs significantly. In agreement with its facultative anaerobic nature, Cfx. aurantiacus in some cases apparently produces structurally different enzymes for heme and BChl biosynthesis, in which one enzyme functions under anoxic conditions and the other performs the same reaction under oxic conditions. The Chl. tepidum mutants produced with modified BChl c and carotenoid species also allow the functions of these pigments to be studied in vivo. PMID- 15340782 TI - Characterisation of a thermoalkali-stable cyclodextrin glycosyltransferase from the anaerobic thermoalkaliphilic bacterium Anaerobranca gottschalkii. AB - The thermoalkaliphilic anaerobic bacterium Anaerobranca gottschalkii produces an extracellular CGTase when grown on starch at 55 degrees C and pH 9.0. The gene encoding this CGTase was cloned and successfully expressed in Escherichia coli. It encodes a protein consisting of 721 amino acids with a signal sequence of 34 amino acids. On SDS-polyacrylamide gels, the purified CGTase from A. gottschalkii displayed the expected molecular mass of 78 kDa. The recombinant enzyme was purified with a yield of 13.5% and displayed a specific activity of 210 units/mg. This CGTase, which represents the first report of a CGTase from an anaerobic thermoalkaliphile, was active at a broad range of temperature and pH, namely 55 70 degrees C and pH 5-10. It completely converted amylose, amylopectin and native starch to cyclodextrins, preferentially alpha-cyclodextrin. With a longer incubation period, the alpha-cyclodextrin to beta-cyclodextrin ratio declined. Variations in substrate type and concentration influenced the product pattern. Increasing the substrate concentration (0.5-20.0%) and glucans containing branching points (alpha-1,6 glycosidic linkages) shifted the product pattern to: beta-cyclodextin > alpha-cyclodextrin > gamma-cyclodextrin. In addition to these cyclodextrins, larger cyclodextrins (>8 glucose units) were formed in the initial reaction period. The CGTase was stabilised against thermal inactivation by calcium ions and high substrate concentrations; and 5 mM of CaCl(2) shifted the apparent melting point of the enzyme from 60 degrees C to 69 degrees C. PMID- 15340783 TI - Studies on the biodegradability of polythioester copolymers and homopolymers by polyhydroxyalkanoate (PHA)-degrading bacteria and PHA depolymerases. AB - The biodegradability of microbial polythioesters (PTEs), a novel class of biopolymers which were discovered recently and can be produced by polyhydroxyalkanoate (PHA)-accumulating bacteria, was studied. Using poly(3 hydroxybutyrate- co-3-mercaptopropionate) [poly(3HB- co-3MP)] as sole carbon source for screening, 22 new bacterial strains were isolated and characterized. Interestingly, none of the PHA-degrading bacteria was able to utilize the homopolymer poly(3MP) as a carbon source for growth or to form clear zones on poly(3MP)-containing agar plates. The extracellular PHA depolymerases from two strains ( Schlegelella thermodepolymerans, Pseudomonas indica K2) were purified to electrophoretic homogeneity and biochemically characterized. The PHA depolymerase of S. thermodepolymerans exhibited a temperate optimum of about 75 degrees C to 80 degrees C and was stable at 70 degrees C for more than 24 h. Regarding the substrate specificities of the PHA depolymerase of S. thermodepolymerans, enzyme activities decreased significantly with increasing 3MP content of the copolymer substrates. Interestingly, no activity could be detected with homoPTEs consisting only of 3MP or of 3-mercaptobutyrate. Similar results were obtained with the PHA depolymerases PhaZ2, PhaZ5 and PhaZ7 of Paucimonas lemoignei which were also investigated. The PHA depolymerase of Ps. indica K2 did not cleave any of the investigated polymers containing 3MP. Gas chromatography, infrared and (1)H-NMR spectrometry and matrix-assisted laser desorption/ionization time-of-flight analysis revealed that 3MPs containing oligomers were enriched in the water-insoluble fraction remaining after partial digestion of poly(3HB- co-3MP) by purified poly(3HB) depolymerase of S. thermodepolymerans. In contrast, 3HB was enriched in the water-soluble fraction, which also contained 3HB- co-3MP dimer obtained by partial digestion of this copolymer by the enzyme. This study clearly indicates that PHA depolymerases are specific for oxoester linkages of PHAs and that the thioester bonds of PTEs cannot be cleaved by this type of enzyme. PMID- 15340785 TI - Desulfosporomusa polytropa gen. nov., sp. nov., a novel sulfate-reducing bacterium from sediments of an oligotrophic lake. AB - Five strains of sulfate-reducing bacteria were isolated from the highest positive dilutions of a most probable number (MPN) series supplemented with lactate and inoculated with sediments from the oligotrophic Lake Stechlin. The isolates were endospore-forming and were motile by means of laterally inserted flagella. They stained Gram-negative and contained b-type cytochromes. CO difference spectra indicated the presence of P582 as a sulfite reductase. Phylogenetic analyses of the 16S rDNA sequences revealed that the isolates were very closely affiliated with the genus Sporomusa. However, sulfate and amorphous Fe(OH)(3), but not sulfite, elemental sulfur, MnO(2), or nitrate were used as terminal electron acceptors. Homoacetogenic growth was found with H(2)/CO(2) gas mixture, formate, methanol, ethanol, and methoxylated aromatic compounds. The strains grew autotrophically with H(2) plus CO(2) in the presence or absence of sulfate. Formate, butyrate, several alcohols, organic acids, carbohydrates, some amino acids, choline, and betaine were also utilized as substrates. The growth yield with lactate and sulfate as substrate was 7.0 g dry mass/mol lactate and thus two times higher than in sulfate-free fermenting cultures. All isolates were able to grow in a temperature range of 4-37 degrees C. Physiologically and by the presence of a Gram-negative cell wall, the new isolates resemble known Desulfosporosinus species. However, phylogenetically they are affiliated with the Gram-negative genus Sporomusa belonging to the Selenomonas subgroup of the Firmicutes. Therefore, the new isolates reveal a new phylogenetic lineage of sulfate-reducing bacteria. A new genus and species, Desulfosporomusa polytropa gen. nov., sp. nov. is proposed. PMID- 15340786 TI - Unusual ADP-forming acetyl-coenzyme A synthetases from the mesophilic halophilic euryarchaeon Haloarcula marismortui and from the hyperthermophilic crenarchaeon Pyrobaculum aerophilum. AB - ADP-forming acetyl-CoA synthetase (ACD), the novel enzyme of acetate formation and energy conservation in archaea Acety - CoA + ADP + Pi<==>acetate + ATP CoA), has been studied only in few hyperthermophilic euryarchaea. Here, we report the characterization of two ACDs with unique molecular and catalytic features, from the mesophilic euryarchaeon Haloarcula marismortui and from the hyperthermophilic crenarchaeon Pyrobaculum aerophilum. ACD from H. marismortui was purified and characterized as a salt-dependent, mesophilic ACD of homodimeric structure (166 kDa). The encoding gene was identified in the partially sequenced genome of H. marismortui and functionally expressed in Escherichia coli. The recombinant enzyme was reactivated from inclusion bodies following solubilization and refolding in the presence of salts. The ACD catalyzed the reversible ADP- and Pi dependent conversion of acetyl-CoA to acetate. In addition to acetate, propionate, butyrate, and branched-chain acids (isobutyrate, isovalerate) were accepted as substrates, rather than the aromatic acids, phenylacetate and indol-3 acetate. In the genome of P. aerophilum, the ORFs PAE3250 and PAE 3249, which code for alpha and beta subunits of an ACD, overlap each other by 1 bp, indicating a novel gene organization among identified ACDs. The two ORFs were separately expressed in E. coli and the recombinant subunits alpha (50 kDa) and beta (28 kDa) were in-vitro reconstituted to an active heterooligomeric protein of high thermostability. The first crenarchaeal ACD showed the broadest substrate spectrum of all known ACDs, catalyzing the conversion of acetyl-CoA, isobutyryl CoA, and phenylacetyl-CoA at high rates. In contrast, the conversion of phenylacetyl-CoA in euryarchaeota is catalyzed by specific ACD isoenzymes. PMID- 15340787 TI - Escherichia coli Hmp, an "oxygen-binding flavohaemoprotein", produces superoxide anion and self-destructs. AB - Escherichia coli Hmp is a homologue of Ralstonia eutropha FHP, the first reported bacterial flavohaemoglobin, and functions in NO detoxification. Photolysis of CO ligated Hmp in the presence of oxygen gave a photodissociable oxy species with k(on) 2.82x10(7) M(-1) s(-1) and k(off) 4.49x10(3) s(-1). The dissociation constant of the primary O(2) compound was 160 microM (25 degrees C, pH 7.0). In order to detect superoxide formation, ferric horseradish peroxidase was used. Hmp formed the oxy compound within milliseconds, followed by formation of compound III, arising from superoxide formation. The rate of superoxide formation was independent of oxygen concentration between 0.05 and 0.7 mM oxygen, suggesting a K(m) <0.05 mM. During prolonged oxidation of NADH, the spectral signals of Hmp decayed and iron was released in a process prevented by superoxide dismutase or catalase. NADH oxidation by purified Hmp was characterised by progressive slowing of oxygen uptake. Inclusion of NO, superoxide dismutase or catalase during NADH oxidation partially protected oxygen uptake, consistent with the formation, in the absence of NO, of reactive oxygen species that inhibit Hmp function. The results are discussed in relation to the tight control exerted on Hmp synthesis in vivo. PMID- 15340788 TI - Sporulation genes in members of the low G+C Gram-type-positive phylogenetic branch ( Firmicutes). AB - Endospore formation is a specific property found within bacteria belonging to the Gram-type-positive low G+C mol% branch ( Firmicutes) of a phylogenetic tree based on 16S rRNA genes. Within the Gram-type-positive bacteria, endospore-formers and species without observed spore formation are widely intermingled. In the present study, a previously reported experimental method (PCR and Southern hybridization assays) and analysis of genome sequences from 52 bacteria and archaea representing sporulating, non-spore-forming, and asporogenic species were used to distinguish non-spore-forming (void of the majority of sporulation-specific genes) from asporogenic (contain the majority of sporulation-specific genes) bacteria. Several sporulating species lacked sequences similar to those of Bacillus subtilis sporulation genes. For some of the genes thought to be sporulation specific, sequences with weak similarity were identified in non-spore forming bacteria outside of the Gram-type-positive phylogenetic branch and in archaea, rendering these genes unsuitable for the intended classification into sporulating, asporogenic, and non-spore-forming species. The obtained results raise questions regarding the evolution of sporulation among the Firmicutes. PMID- 15340789 TI - Continuing opportunities for general microbiology. PMID- 15340790 TI - Ferrihydrite reduction by Geobacter species is stimulated by secondary bacteria. AB - Geobacter species such as G. bremensis, G. pelophilus, and G. sulfurreducens are obligately anaerobic and grow in anoxic, non-reduced medium by fast reduction of soluble ferric citrate. In contrast, insoluble ferrihydrite was either only slowly or not reduced when supplied as electron acceptor in similar growth experiments. Ferrihydrite reduction was stimulated by addition of a reducing agent or by concomitant growth of secondary bacteria that were physiologically and phylogenetically as diverse as Escherichia coli, Lactococcus lactis, or Pseudomonas stutzeri. In control experiments with heat-inactivated Geobacter cells and viable secondary bacteria, no ( E. coli, P. stutzeri) or only little ( L. lactis) ferrihydrite was reduced. Redox indicator dyes showed that growing E. coli, P. stutzeri, or L. lactis cells lowered the redox potential of the medium in a similar way as a reducing agent did. The lowered redox potential was presumably the key factor that stimulated ferrihydrite reduction by all three Geobacter species. The observed differences in anoxic non-reduced medium with ferric citrate versus ferrihydrite as electron acceptor indicated that reduction of these electron acceptors involved different cellular components or different biochemical strategies. Furthermore, it appears that redox-sensitive components are involved, and/or that gene expression of components needed for ferrihydrite reduction is controlled by the redox state. PMID- 15340791 TI - Thermotolerant poly(3-hydroxybutyrate)-degrading bacteria from hot compost and characterization of the PHB depolymerase of Schlegelella sp. KB1a. AB - Eighteen gram-negative thermotolerant poly(3-hydroxybutyrate) (PHB)-degrading bacterial isolates ( T(max) approximately 60 degrees C) were obtained from compost. Isolates produced clearing zones on opaque PHB agar, indicating the presence of extracellular PHB depolymerases. Comparison of physiological characteristics and determination of 16S rRNA gene sequences of four selected isolates revealed a close relatedness of three isolates (SA8, SA1, and KA1) to each other and to Schlegelella thermodepolymerans and Caenibacterium thermophilum. The fourth strain, isolate KB1a, showed reduced similarities to the above-mentioned isolates and species and might represent a new species of Schlegelella. Evidence is provided that S. thermodepolymerans and C. thermophilum are only one species. The PHB depolymerase gene, phaZ, of isolate KB1a was cloned and functionally expressed in Escherichia coli. Purified PHB depolymerase was most active around pH 10 and 76 degrees C. The DNA-deduced amino acid sequence of the mature protein (49.4 kDa) shared significant homologies to other extracellular PHB depolymerases with a domain substructure: catalytic domain type 2-linker domain fibronectin type 3-substrate-binding domain type 1. A catalytic triad consisting of S(20), D(104), and H(138) and a pentapeptide sequence (GLS(20)AG) characteristic for PHB depolymerases (PHB depolymerase box, GLSXG) and for other serine hydrolases (lipase box, GXSXG) were identified. PMID- 15340792 TI - The role of the sulfur globule proteins of Allochromatium vinosum: mutagenesis of the sulfur globule protein genes and expression studies by real-time RT-PCR. AB - During oxidation of reduced sulfur compounds, the purple sulfur bacterium Allochromatium vinosum stores sulfur in the periplasm in the form of intracellular sulfur globules. The sulfur in the globules is enclosed by a protein envelope that consists of the homologous 10.5-kDa proteins SgpA and SgpB and the smaller 8.5-kDa SgpC. Reporter gene fusions of sgpA and alkaline phosphatase showed the constitutive expression of sgpA in A. vinosum and yielded additional evidence for the periplasmic localization of the sulfur globules. Expression analysis of the wild-type sgp genes by quantitative RT-PCR using the LightCycler system showed the constitutive expression of all three sgp genes. The expression of sgpB and sgpC is significantly enhanced under photolithotrophic conditions. Interestingly, sgpB is expressed ten times less than sgpA and sgpC implying that SgpA and SgpC are the "main proteins" of the sulfur globule envelope. Mutants with inactivated sgpA or sgpB did not show any differences in comparison with the wild-type, i.e., the encoded proteins can replace each other, whereas inactivation of sgpC leads to the formation of considerably smaller sulfur globules. This indicates a role of SgpC for globule expansion. A sgpBC double mutant was unable to grow on sulfide and could not form sulfur globules, showing that the protein envelope is indispensible for the formation and deposition of intracellular sulfur. PMID- 15340793 TI - Microorganisms degrading chlorobenzene via a meta-cleavage pathway harbor highly similar chlorocatechol 2,3-dioxygenase-encoding gene clusters. AB - Pseudomonas putida GJ31 harbors a degradative pathway for chlorobenzene via meta cleavage of 3-chlorocatechol. Pseudomonads using this route for chlorobenzene degradation, which was previously thought to be generally unproductive, were isolated from various contaminated environments of distant locations. The new isolates, Pseudomonas fluorescens SK1 (DSM16274), Pseudomonas veronii 16-6A (DSM16273), Pseudomonas sp. strain MG61 (DSM16272), harbor a chlorocatechol 2,3 dioxygenase (CbzE). The cbzE-like genes were cloned, sequenced, and expressed from the isolates and a mixed culture. The chlorocatechol 2,3-dioxygenases shared 97% identical amino acids with CbzE from strain GJ31, forming a distinct family of catechol 2,3-dioxygenases. The chlorocatechol 2,3-dioxygenase, purified from chlorobenzene-grown cells of strain SK1, showed an identical N-terminal sequence with the amino acid sequence deduced from cloned cbzE. In all investigated chlorobenzene-degrading strains, cbzT-like genes encoding ferredoxins are located upstream of cbzE. The sequence data indicate that the ferredoxins are identical (one amino acid difference in CbzT of strain 16-6A compared to the others). In addition, the structure of the operon downstream of cbzE is identical in strains GJ31, 16-6A, and SK1 with genes cbzX (unknown function) and the known part of cbzG (2-hydroxymuconic semialdehyde dehydrogenase) and share 100% nucleotide sequence identity with the entire downstream region. The current study suggests that meta-cleavage of 3-chlorocatechol is not an atypical pathway for the degradation of chlorobenzene. PMID- 15340794 TI - The role of the active site-coordinating cysteine residues in the maturation of the H2-sensing [NiFe] hydrogenase from Ralstonia eutropha H16. AB - The H(2)-splitting active site of [NiFe] hydrogenases is tightly bound to the protein matrix via four conserved cysteine residues. In this study, the nickel binding cysteine residues of HoxC, the large subunit of the H(2)-sensing regulatory hydrogenase (RH) from Ralstonia eutropha, were replaced by serine. All four mutant proteins, C60S, C63S, C479S, and C482S, were inactive both in H(2) sensing and H(2) oxidation and did not adopt the native oligomeric structure of the RH. Nickel was bound only to the C482S derivative. The assembly of the [NiFe] active site is a complex process that requires the function of at least six accessory proteins. Among these proteins, HypC has been shown to act as a chaperone for the large subunit during the maturation process. Immunoblot analysis revealed the presence of a strong RH-dependent HypC-specific complex in extracts containing the C60S, C63S, and C482S derivatives, pointing to a block in maturation for these mutant proteins. The lack of this complex in the extract containing C479S indicates that this specific cysteine residue might be crucial for the interaction between HoxC and HypC. PMID- 15340795 TI - Sulfoacetate generated by Rhodopseudomonas palustris from taurine. AB - Genes thought to encode (a) the regulator of taurine catabolism under carbon limiting or nitrogen-limiting conditions and (b) taurine dehydrogenase were found in the genome of Rhodopseudomonas palustris. The organism utilized taurine quantitatively as a sole source of nitrogen (but not of carbon) for aerobic and photoheterotrophic growth. No sulfate was released, and the C-sulfonate bond was recovered stoichiometrically as sulfoacetate, which was identified by mass spectrometry. An inducible sulfoacetaldehyde dehydrogenase was detected. R. palustris thus contains a pathway to generate a natural product that was previously believed to be formed solely from sulfoquinovose. PMID- 15340796 TI - F420H2 oxidase (FprA) from Methanobrevibacter arboriphilus, a coenzyme F420 dependent enzyme involved in O2 detoxification. AB - Cell suspensions of Methanobrevibacter arboriphilus catalyzed the reduction of O(2) with H(2) at a maximal specific rate of 0.4 U (micromol/min) per mg protein with an apparent K(m) for O(2) of 30 microM. The reaction was not inhibited by cyanide. The oxidase activity was traced back to a coenzyme F(420)-dependent enzyme that was purified to apparent homogeneity and that catalyzed the oxidation of 2 F(420)H(2) with 1 O(2) to 2 F(420) and 2 H(2)O. The apparent K(m) for F(420) was 30 microM and that for O(2) was 2 microM with a V(max) of 240 U/mg at 37 degrees C and pH 7.6, the pH optimum of the oxidase. The enzyme did not use NADH or NADPH as electron donor or H(2)O(2) as electron acceptor and was not inhibited by cyanide. The 45-kDa protein, whose gene was cloned and sequenced, contained 1 FMN per mol and harbored a binuclear iron center as indicated by the sequence motif H-X-E-X-D-X(62)-H-X(18)-D-X(60)-H. Sequence comparisons revealed that the F(420)H(2) oxidase from M. arboriphilus is phylogenetically closely related to FprA from Methanothermobacter marburgensis (71% sequence identity), a 45-kDa flavoprotein of hitherto unknown function, and to A-type flavoproteins from bacteria (30-40%), which all have dioxygen reductase activity. With heterologously produced FprA from M. marburgensis it is shown that this protein is also a highly efficient F(420)H(2) oxidase and that it contains 1 FMN and 2 iron atoms. The presence of F(420)H(2) oxidase in methanogenic archaea may explain why some methanogens, e.g., the Methanobrevibacter species in the termite hindgut, cannot only tolerate but thrive under microoxic conditions. PMID- 15340797 TI - Overproduction of NAD+ and 5'-inosine monophosphate in the presence of 10 microM Mn2+ by a mutant of Corynebacterium ammoniagenes with thermosensitive nucleotide reduction (nrd(ts)) after temperature shift. AB - Corynebacterium ammoniagenes strain CH31 is thermosensitive due to a mutation in nucleotide reduction ( nrd(ts)). The strain was examined for nucleotide overproduction upon shifting the culture temperature to a range of elevated temperatures. No overproduction of NAD(+) was detected in the control maintained at 27 degrees C whereas NAD(+) was accumulated extracellularily by strain CH31 at 37 degrees C and at 40 degrees C. As a result of the temperature shift, division inhibited cells displayed only limited elongation. This is a characteristic morphological feature of cell-cycle-arrested coryneform bacteria. Ribonucleotide reductase (RNR) activity was inactivated immediately after the temperature shift in the NAD(+)-proficient cultures, leading presumably to an exhaustion of deoxyribonucleotide pools and impairment of DNA replication. In contrast to the low extracellular accumulation of NAD(+), at the non-permissive temperature of 35 degrees C a distinct capacity for intracellular nucleotide overproduction was revealed by a new method using nucleotide-permeable cells. The approach of shifting the culture temperature was applied successfully to the overproduction of taste-enhancing nucleotides in the presence of 10 microM Mn(2+). Concomitant with a dramatic loss of viability, the thermosensitive mutant CH31 accumulated 5.3 g 5'-inosine monophosphate per liter following the addition of hypoxanthine as precursor for the salvage pathway. PMID- 15340798 TI - Identification of a regulatory pathway that controls the heavy-metal resistance system Czc via promoter czcNp in Ralstonia metallidurans. AB - The CzcCBA cation-proton-antiporter is the most complicated and efficient heavy metal resistance system known today and is essential for survival of Ralstonia metallidurans at high cobalt, zinc, or cadmium concentrations. Expression of Czc is tightly controlled by the complex interaction of several regulators. Double- and multiple-deletion studies demonstrated that four regulators encoded downstream of the czcCBA operon, CzcD, CzcS, CzcR and the newly identified CzcE, are involved in, but not essential for metal-dependent induction of czc. These proteins control expression of the czcNICBA region from the promoter czcNp. Northern analysis showed that czcDRS was transcribed as czcDR-specific and czcDRS specific mRNAs. Transcription of czcE occurred independently of czcDRS transcription and was induced by zinc. CzcE is a periplasmic protein as indicated by phoA fusions. CzcE was purified and identified as a metal-binding protein. These data demonstrate that the transport protein CzcD, the two-component regulatory system CzcR, CzcS, and the periplasmic metal-binding protein CzcE exert metal-dependent control of czcNICBA expression via regulation of czcNp activity. PMID- 15340799 TI - Health care utilization and expenditures in the United States: a study of osteoporosis-related fractures. AB - BACKGROUND: More than 1.5 million fractures occur due to osteoporosis each year. This study examines the annual health care utilization and associated expenditures of osteoporotic patients who sustain a new fragility fracture and of those without a new fracture. METHODS AND PROCEDURES: The study sample from commercial claims databases consisted of patients enrolled in US plans between January 1, 1997, and December 31, 2001. Patients with both an osteoporosis diagnosis and a related fracture were classified as "osteoporosis with concurrent fracture"; all other osteoporosis patients were classified as "osteoporosis without concurrent fracture." Annual utilization and expenditures for the concurrent-fracture cohort were compared with those without concurrent fracture, as well as with a group of patients without osteoporosis (controls) that was matched to the concurrent-fracture cohort based on age, gender, US region, health plan type, and length of enrollment. Exponential conditional mean models were used to compute regression-adjusted total expenditures across the groups. The differences in adjusted expenditures were used to generate the economic burden-of illness estimates. RESULTS: Osteoporosis patients with concurrent fracture incurred more than twice the overall health care expenditures in the study period, compared with those without fracture (US $15,942 vs $6,476), and nearly three times those of the control group (US $15,942 vs $4,658). Approximately 25% of the overall health care expenditures (US $4,014 of $15,942) for the concurrent fracture group were osteoporosis-related expenditures, leading to the conclusion that comorbid conditions in osteoporosis patients with concurrent fracture contribute significantly to overall health care costs. Some of these comorbidity related costs were likely due to pain-related disorders, which occurred significantly more frequently in the concurrent-fracture cohort than in the other groups. CONCLUSION: Osteoporosis-related expenditures, particularly those related to fracture, were substantial. However, non-osteoporosis-related expenditures to treat comorbid conditions constituted 75% of the overall health care costs in the year after an osteoporosis-related fracture, which warrants further investigation. PMID- 15340800 TI - Long-term prediction of three-dimensional bone architecture in simulations of pre , peri- and post-menopausal microstructural bone remodeling. AB - The mechanical behavior of trabecular bone depends on the internal bone structure. It is generally accepted now that the trabecular bone structure is a result of a load adaptive bone remodeling. The mathematical laws that relate bone remodeling to the local state of stress and strain, however, are still under investigation. The aim of this project was to investigate if changes in the trabecular architecture as observed with age-related bone loss and osteoporosis can be predicted from a computer model that simulates bone resorption after hormone depletion based on realistic models of trabecular microstructure using micro-computed tomography (muCT). A compact desktop muCT providing a nominal isotropic resolution of 14 mum was used to measure two groups of seven trabecular bone specimens from pre-menopausal and post-menopausal women respectively. A novel algorithm was developed to simulate age-related bone loss for the specimens in the first group. The algorithm, also referred to as simulated bone atrophy (SIBA), describes a truly three-dimensional approach and is based directly on cellular bone remodeling with an underlying realistic time frame. Bone resorption is controlled by osteoclastic penetration depth and bone formation is governed by the efficiency level of the osteoblasts. The simulation itself describes an iterative process with a cellular remodeling cycle of 197 days. Activation frequency is controllable and can be adjusted for the different phases of pre-, peri- and post-menopause. For our simulations, osteoblastic and osteoclastic activities were in balance until the onset of menopause, set to be at the age of 50 years. In that period, the structure remained almost constant. After the onset of menopause an imbalance in the cell activities was modeled resulting in a net bone loss. The doubling of the activation frequency in the peri-menopausal phase caused a pronounced loss. Using advanced animation tools and quantitative bone morphometry, the changes in bone architecture associated with the bone loss were monitored over an average observation time of 43 years until the age of 80 years. In that time, bone volume density decreased monotonously with the progression of the simulation for all specimens. Right after the onset of menopause, bone was lost fast, where with the progression of age losses slowed down. The structures at the end-point of the simulations were then compared qualitatively and quantitatively to the structures of the post-menopausal group with all morphometric indices being within a narrow margin of error. These results suggest the feasibility of transforming "normal" to "osteopenic" bone on a microstructural level yielding in realistic bone models similar in appearance as well as in structural behavior if compared to a post-menopausal group of women. PMID- 15340801 TI - Clinical utility of laboratory testing in women with osteoporosis. AB - Clinicians regularly order laboratory tests in women with osteoporosis to assess if an underlying medical condition is contributing to bone loss. To determine which laboratory tests are associated with osteoporosis we conducted a secondary analyses of data collected as part of the Fracture Intervention Trial (FIT), which included 15,316 postmenopausal women. Women had tests of liver and kidney function, mineral metabolism, electrolytes and complete blood count, femoral neck and total body BMD and spinal radiographs. The prevalence of abnormal tests in women with osteoporosis compared to women without was not different, except for low TSH (<0.5 IU/ml). Among women with and without osteoporosis at the femoral neck the prevalence of low TSH was 4.9% (95% CI: 4.4-5.5) and 3.7% (95% CI: 3.3 4.1), respectively, yielding a positive likelihood ratio of 1.2 (95% CI: 1.1 1.3). We observed similar results for women with vertebral fractures compared to women without vertebral fractures; positive likelihood ratio of 1.4 (95% CI: 1.3 1.6). Our data suggests that when assessing healthy women with osteoporosis ordering a routine panel of laboratory tests is not useful but measuring TSH may be informative. PMID- 15340802 TI - PelviSoft BioMesh augmentation of rectocele repair: the initial clinical experience in 35 patients. AB - Graft materials have been utilized in the repair of posterior vaginal wall defects to enhance anatomical and functional results, and to improve long-term outcomes. We report on our initial series of 35 patients treated with porcine dermal acellular collagen matrix BioMesh (PelviSoft BioMesh, CR Bard, Cranston, R.I., USA), which has alleviated problems with early postoperative vaginal mucosal dehiscence and delayed healing experienced with the use of other graft materials in the posterior vaginal wall. PMID- 15340803 TI - [Experiences with six different intervertebral disc spacers for spondylodesis of the cervical spine]. AB - Spondylodesis of the cervical spine with autologous bone in cases of degenerative disease is associated with pain and complications in the os ilium region. Can this problem be solved when using intervertebral disc spacers? Between September 1997 and January 2003, 267 patients with degenerative disease of the cervical spine were treated operatively and monitored in a follow-up examination. In osteochondrosis of the cervical spine and/or disc herniation, we performed a discectomy using a ventral approach and removed dorsal osteophytes with a high speed drill. The choice of the six different spacers was randomized. We used 66 titanium disc spacers manufactured by Aesculap, 54 titanium disc spacers by Weber, 52 titanium disc spacers by Intromed, 18 Wing spacers by Medinorm, 17 carbonium spacers by AcroMed, and 50 PEEK spacers by Intromed and did follow-up examinations after 3 and 12 months, respectively. The titanium spacer manufactured by Weber is characterized by good handling and a moderate price. The titanium cage from Intromed has the lowest price and good handling characteristics during operation. The X-ray contrast of the carbonium spacer made by AcroMed is not always sufficient for safe implantation, especially in the lower cervical spine. Handling is good, but the price is too high. The AcoMed, Medinorm, and Intromed titanium spacers show a tendency for penetrating ground plates of adjacent vertebra. The titanium cage Cespace manufactured by Aesculap has good handling characteristics and a moderate price. The PEEK spacer from Intromed can be implanted safely but shows a tendency for dislocation. Long-term results are yet to be reported. PMID- 15340804 TI - Aquaporins and the central nervous system. PMID- 15340805 TI - Reduction in late postoperative pain after iliac crest bonegraft harvesting for cervical fusion: a controlled double-blinded study of 100 patients. AB - BACKGROUND: A randomized prospective double-blinded study was conducted in 100 patients suffering from mono- or bisegmental cervical retrospondylosis or disc herniation. METHOD: In group I, 50 patients were treated by injection of 10 ml Ropivacaine 7,5% at the iliac crest bonegraft donor site. Local anaesthetic (LA) was injected through the wound drainage after closure of the muscle fascia, the suction drainage was opened after closure of the skin. Group II was treated with 0,9% saline. Operator and patient were blinded to the injected substance. Daily controls of pain intensity were made with the 10 cm visual analog scale from 0 (no pain) to 10 (severe pain) for 5 days. All patients were questioned regarding pain character and movement provoking pain. Additional pain medication was standardized. FINDINGS: Statistical analysis of mean pain intensity over the whole hospital stay showed a significant difference in pain intensity between the two groups (p = 0,017, Chi-Square test). The comparison between pain intensity with LA and without LA showed a gradual increase in statistical significance from day 1 to day 5 (day 1: p = 0,54, not significant; day 2: p = 0,026; day 3: p = 0,008; day 4: p = 0,004; day 5: p = 0,002). INTERPRETATION: This data shows that intra-operative blockage of peripheral nociceptive structures results in decreased pain at later time points. We conclude that wound infiltration with 7,5% Ropivacaine after bonegraft removal at the iliac crest is effective in reducing postoperative pain. PMID- 15340806 TI - Considerations in the surgical treatment of superior-wall type aneurysm at the proximal (M1) segment of the middle cerebral artery. AB - OBJECTIVE: To present our experience and analyze the risks of neck clipping for superior-wall type aneurysms of the proximal segment of the middle cerebral artery (M1-Sup aneurysm). METHODS: Of 14 patients with M1-Sup aneurysms, 4 suffered postoperative infarctions in the territory of the lenticulostriate arteries (LSA) or fronto-orbital arteries. We re-examined our intraoperative findings and clinical records in an effort to identify possible causes. RESULTS: The patency of the LSA was confirmed at the end of surgery in all 14 cases. We posit that temporary occlusion of the LSA by a permanent clip resulted in delayed obstruction of the LSA in 3 patients. Other possibilities we considered were relatively long temporary occlusion of the M1, slipping of the clips and twisting of the clip blades after release of the brain retractors. CONCLUSION: M1-Sup aneurysms are some of the most complicated aneurysms; they carry the risk of perforator injury during neck clipping. Surgical considerations to avoid perforator injury are discussed. PMID- 15340807 TI - Anterior interhemispheric approach for distal anterior cerebral artery aneurysm surgery: preoperative analysis of the venous anatomy can help to avoid venous infarction. AB - BACKGROUND: The anterior interhemispheric approach is a well-known operative technique for a distal anterior cerebral artery (ACA) aneurysm. However, a frontal parasagittal bridging vein is occasionally sacrificed in this approach, creating a risk of postoperative venous infarction. METHOD: To evaluate the risk of venous infarction, the current study investigated the venous phase of preoperative angiograms and postoperative CT scans for 20 patients in whom a parasagittal bridging vein had been sacrificed during the anterior interhemispheric approach. The draining territory index of the sacrificed bridging vein (DTISBV), draining territory index of the adjacent collateral ascending draining veins (DTIADV), and drainage impediment index (DII) were all calculated, plus the development of the superficial Sylvian vein (SSV) was evaluated. FINDINGS: Extensive postoperative venous infarction occurred in two of four patients with an aplastic SSV, for whom the DTISBVs were 41% and 43%, and the DIIs 26% and 37%. Among 16 patients with a normal SSV, two patients suffered postoperative venous congestion, and their DTISBVs were 40% and 50%, and their DIIs 21% and 30%. The other 14 patients without any postoperative venous drainage problems had a lower DTISBV and DII, where the DTISBV was 22.3+/-6.8% (mean+/-SD) and the DII 4.3+/-8.1%. CONCLUSIONS: The present results suggest that the venous phase of a preoperative angiogram should be carefully evaluated before distal ACA aneurysm surgery. In particular, a DTISBV and DII over 50% and 30%, respectively, in patients with a normal SSV and over 40% and 25%, respectively, in patients with an aplastic SSV were found to be critical to the production of postoperative venous infarction. PMID- 15340808 TI - Ruptured de novo intracranial aneurysms. AB - OBJECTIVE: We describe the actual state of ruptured de novo intracranial aneurysms to contribute to a guideline of follow-up for the patients with treated intracranial aneurysm. METHODS: The authors retrospectively investigated 12 cases drawn from 483 consecutive cases of aneurysmal subarachnoid hemorrhage at our institute over a period of 22 years, in which a previously undemonstrated (hence "de novo") intracranial saccular aneurysm formed and ruptured after successful treatment of a prior aneurysm. FINDINGS: The 12 cases constitute 2.5% of the 483 patients who left our hospital alive. Eleven cases were females and one was a male with a mean age of 55.7 years (range 29-75) at the first subarachnoid haemorrhage (SAH) and an interval between the first and the second rupture of 10.7 years (range: 2.6-23.8, standard deviation: 6.86, 95% confidence interval: 6.39-15.1). Four cases did not have risk factors such as hypertension, family history, smoking, multiple aneurysms, and moyamoya disease. None of these ruptured de novo aneurysms was at the same location as the original lesion. One third (4 cases) of the de novo lesions in our series were found on the opposite side to each prior lesion. INTERPRETATION: For not only young but also elder patients with a treated aneurysm (from the fifth decade to the sixth), especially for women, late angiography or alternative modalities of less-invasive examination should be considered. To detect de novo intracranial aneurysms before rupture, the search for a de novo aneurysm should be performed within 6.39 years after a previous examination that shows an aneurysm to be nonexistent, in view of the 95% confidence interval of the mean time to de novo aneurysmal rupture (6.39 15.1 years). If applied this survey, 75% (8 cases of 12 cases) of our de novo aneurysms would be detected before rupture. PMID- 15340809 TI - Normal values of volume flow in the internal carotid artery measured by a new angle-independent Doppler technique for evaluating cerebral perfusion. AB - BACKGROUND: Cerebral blood flow is an important parameter when monitoring critically ill patients. Blood flow volume within the internal carotid artery (ICA) was shown to be correlated with the cerebral blood flow. The aim of our study was to provide normal values of the internal carotid artery volume flow using this new technology (QuantixND, Cardiosonix Ltd., Israel). METHOD: The QuantixND System is an angle-independent Doppler system that employs two digital high resolution ultrasound heads within one probe in an defined angle to each other. Thus several flow velocities within the vessel and the vessel diameter can be measured. 77 healthy patients (41 women, 36 men) were included and divided into age groups of ten-year intervals (mean age 48.9 years). Internal carotid artery flow was evaluated as well as physiological and hematological parameters (hematocrit, arterial blood pressure etc). FINDINGS: We found that the blood flow volume in the ICA decreased significantly with age. No side-to-side effects as well as no gender-related differences could be observed. There was no influence of hemoglobin, hematocrit and blood pressure in this healthy population. CONCLUSIONS: There is an age-related decrease in blood flow volume with age that can be easily and exactly determined by the use of the new angle-independent Doppler technique. PMID- 15340810 TI - Repeated gamma knife radiosurgery for multiple metastatic brain tumours. AB - BACKGROUND: The effectiveness of repeated gamma knife radiosurgery (GKS) for the treatment of multiple metastatic brain tumours was evaluated. METHODS: This study included 16 patients with 242 tumours, 10 men and 6 women with a mean age of 60.3 years at initial GKS, who underwent GKS four times or more for newly developed metastatic tumours. FINDINGS: Sixteen patients underwent a total of 83 GKS procedures (range 4 to 8, mean 5.2). The mean number of metastases at each GKS procedure was 2.9 and the number of tumours tended to increase at the 5th GKS procedure compared with the 1st, but not significantly. The mean interval between each procedure was 4.8 months and was not significantly different. Median survival was 22.4 months (range 9.4-78.9 months) and the primary site was not correlated with survival time. The total number of treated tumours tended to correlate to survival time, but not significantly. Use of adjuvant whole brain radiation also had no significant effect on survival time. The Karnofsky performance status was maintained at more than 70 in most patients, but decreased significantly between initial and final GKS. Death due to progression of brain lesions occurred in only about 30% of patients regardless of the multiple newly developed brain metastases. CONCLUSIONS: Repeated radiosurgery for brain metastases is effective and relatively long survival can be expected in some patients associated with a low risk of radiation-induced injury. PMID- 15340811 TI - Craniotomies without burr holes using an oscillating saw. AB - OBJECTIVE: The object of this study is to describe a new method for performing craniotomies which obviates the need for burr holes, improves bony reconstruction, and reduces post-operative cosmetic deformities. Moreover, this technique provides excellent exposure of skull base structures and dural venous sinuses. METHODS: Craniotomies in varied locations are created with the use of a micro-oscillating saw and chisel. No burr holes are used and reconstruction with plates and screws is unnecessary. RESULTS: We initially applied our technique to approaches to the anterior skull base with combined craniofacial tumour resections. We have since performed over 2000 craniotomies of any size and shape in all supratentorial locations using the oscillating saw. CONCLUSIONS: We have found that our method creates better cosmetic results than standard techniques and is safer for craniotomies spanning dural venous sinuses. With experience, operating time was significantly reduced and costs were lowered because reconstruction with fixation devices was not needed. PMID- 15340812 TI - Is idiopathic normal pressure hydrocephalus an independent entity? PMID- 15340813 TI - The ELANA technique: constructing a high flow bypass using a non-occlusive anastomosis on the ICA and a conventional anastomosis on the SCA in the treatment of a fusiform giant basilar trunk aneurysm. AB - A patient with a partially thrombosed fusiform giant basilar trunk aneurysm presented with devastating headache and symptoms of progressive brain stem compression. Having an aneurysm inaccessible for endovascular treatment, and after failing a vertebral artery balloon occlusion test, he was offered bypass surgery in order to exclude the aneurysm from the cerebral circulation and relieve his symptoms. A connection between the intracranial internal carotid artery and the superior cerebellar artery was created whereupon the basilar artery was ligated just distally to the aneurysm. The proximal anastomosis on the internal carotid artery was made using the excimer laser-assisted non-occlusive anastomosis (ELANA) technique, while a conventional end-to-side anastomosis was used for the distal anastomosis on the superior cerebellar artery. Intra operative flowmetry showed a flow through the bypass of 40 ml/min after ligation of the basilar artery. An angiogram 24 hours later showed normal filling of the bypass and the vessels supplied by it, but also disclosed a subtotal occlusion of the proximal ipsilateral middle cerebral artery with delayed filling distally. The patient, who had a known thrombogenic coagulopathy, died the following day. Autopsy showed no signs of ischemia in the territories supplied by the bypass, but a thrombus in the proximal middle cerebral artery and massive acute hemorrhagic infarction with swelling in its territory and uncal herniation. Multiple fresh thrombi were found in the lungs. The ELANA anastomosis showed re endothelialisation without thrombus formation on the inside. PMID- 15340814 TI - Surgical treatment of seizures from the peri-Sylvian area by perinatal insult: a case report of ictal hypersalivation. AB - OBJECTIVES AND IMPORTANCE: It is important to evaluate the seizure manifestation of epilepsy before surgical planning. A patient with partial epilepsy manifesting hypersalivation who underwent resection of the epileptogenic foci with satisfactory postoperative seizure control is reported. CLINICAL PRESENTATION AND INTERVENTION: A 26-year-old man, with a history of perinatal asphyxia, started having medically intractable partial epilepsy at the age of 10 years. His seizure was characterized by throat discomfort followed by hypersalivation. Brain MRI showed an atrophic lesion around the peri-Sylvian area. Scalp recorded EEG did not demonstrate robust epileptiform activity localized enough to define the epileptogenic zone. The patient underwent invasive recording by multiple subdural electrode grids, which showed that the seizure arose from the left anterior frontal operculum. After resection of epileptogenic opercular cortex, the seizures disappeared with no additional neurological deficits. CONCLUSION: Although the responsible sites for ictal drooling are distributed in multiple areas including insula, medial temporal area and operculum, the seizure can be successfully controlled by focus resection of the frontal opercular area in a selected patient with careful presurgical evaluation. PMID- 15340815 TI - Obsessive-compulsive disorder after unilateral caudate nucleus bleeding. AB - Obsessive compulsive disorders (OCD) may be encountered after basal ganglia lesions of various aetiologies. These lesions are usually bilateral. We report here the case of a 24 years old man who developed a pure compulsive behavior after a unilateral left-sided caudate nucleus hemorrhage due to a cavernoma. The pathophysiology of this compulsive disorder probably reflects a frontal cortex deafferentation mechanism. Behavioral, psychological and medical (serotoninergic) treatments are usually proposed but the efficacy of such therapy remains to be investigated in secondary OCD. PMID- 15340816 TI - Atypical teratoid/rhabdoid tumour with leptomeningeal dissemination in an adult. AB - A 22-year-old man presented with a rare case of atypical teratoid/rhabdoid tumour (AT/RT). Magnetic resonance imaging showed a left cerebellar mass with leptomeningeal dissemination. Partial resection was performed. Histological examination revealed AT/RT. Postoperatively, whole neuraxis and local irradiation were performed. Three-drug chemotherapy with ifosfamide, cisplatin, and etoposide, and adjuvant intrathecal administration of methotrexate were repeated. Near complete response was achieved, and no tumour recurrence/progression has been noticed during the follow up of 24 months. Intensive radiochemotherapy can successfully control AT/RT, even with leptomeningeal dissemination. PMID- 15340817 TI - Postoperative pseudoaneurysm of the superficial temporal artery (S.T.A.) treated with Thrombostat (thrombin glue) injection. AB - BACKGROUND: Pseudo-aneurysm is a rare complication of craniotomy. Blunt injury to the temporal artery region is the usual cause, but still a rare complication. CLINICAL PRESENTATION: A patient with subarachnoid hemorrhage was successfully treated by aneurysm clipping. The patient developed hydrocephalus, and was admitted for a shunt operation seventeen days later. The craniotomy had healed normally, but a palpable temporal lump was present in the skin incision. INTERVENTION: The pulsating mass proved to be a postoperative aneurysm of the superficial temporal artery (S.T.A.) and was successfully occluded with 500 units Thrombostat (thrombin glue) which was injected into the aneurysm sac using a 22 gauge needle guided by ultrasound. The permanency of the obliteration was verified by ultrasound examination. PMID- 15340818 TI - Pure distant, leptomeningeal metastasis of esthesioneuro-epithelioma. AB - BACKGROUND: Esthesioneuro-epithelioma is a very rare variant of olfactory neuroblastoma that originates in the region of the cribriform plate. Its intracranial manifestation is due to infiltration of the anterior skull base and frontal lobes. METHODS: The authors describe a 52-year-old man with a history of a subtotally resected right maxillary sinus ethesioneuro-epithelioma 2 years earlier who presented with a left hemiparesis and signs of increased intracranial pressure. CT examination revealed a tumour of the right temporoparietal region. The tumour and its dural attachment were totally removed. The patient made a full recovery postoperatively. FINDINGS: The features of esthesioneuro-epithelioma were found on histopathological examination. INTERPRETATION: The intracranial tumour resected was a distant, leptomeningeal metastasis of esthesioneuro epithelioma. To our knowledge such a documented case has not been published before. PMID- 15340819 TI - Exophytic ependymomas of the spinal cord. AB - Intramedullary ependymomas of the spinal cord with exophytic components are rare outside the filum or conus region. Two cases of combined intradural intramedullary and extramedullary ependymomas of the spinal cord are presented. At operation, the tumours proved to be primarily intramedullary but had a contiguous exophytic component that extruded either through a defect in the ventral pia to encase the anterior spinal artery in one patient, or through the dorsal root entry zone in the second patient. When removing intramedullary spinal tumours with an exophytic component, separate removal of the intramedullary and extramedullary components is recommended, rather than en bloc resection, to prevent possible injury to the vascular supply of the spinal cord. PMID- 15340820 TI - Primary cervical spinal epidural Extra-osseous Ewing's sarcoma. AB - Cases of primary spinal epidural Extra-osseous Ewing's sarcoma (EES) are rarely seen and a good prognosis for EES cannot be expected since a high incidence of local recurrence and metastasis frequently occur. We present a case of cervical spinal epidural EES in a 7-year-old girl with long survival after tumour resection. She also received adjuvant treatment with peripheral blood stem cell transplantation (PBSCT). This management successfully improved her condition and she was in complete remission without neurological deficit 60 months after surgery. Such a good prognosis of EES in the cervical spine is very rare. It is likely that magnetic resonance imaging (MRI) lead to a diagnosis of this rare type of EES at an early stage of the disease. PBSCT could be useful as an adjuvant to prolong the period of complete remission. PMID- 15340821 TI - A knot in the catheter--an unusual cause of ventriculo-peritoneal shunt blockage. AB - A 25-year-old woman, who was 25 weeks pregnant, underwent insertion of a VP shunt for hydrocephalus, secondary to a bithalamic glioma. Two months later, she represented with symptoms of raised intracranial pressure and MR scan revealed increased ventricular size. On exploration of the shunt, manometry with saline confirmed blockage of the catheter distal to the valve. On re-opening the abdominal wound, the peritoneal catheter was found to be knotted, 2 cm from the end. This segment of the catheter was replaced, with resolution of symptoms, post operatively. The present case illustrates that a knot in the peritoneal catheter is an extremely rare cause of shunt malfunction. Possible mechanisms underlying it are discussed. PMID- 15340822 TI - Decompressive craniectomy with lattice duraplasty. PMID- 15340823 TI - Ventral brainstem enterogenous cyst. PMID- 15340826 TI - Expression of fibronectin and TGF-beta1 mRNA and protein suggest altered regulation of extracellular matrix in degenerated disc tissue. AB - We studied the distribution of fibronectin (a marker for "active" reparative connective tissue processes) and TGF-beta1 (a cytokine controlling the connective tissue metabolism) in intervertebral disc tissue from individuals of different age and various histomorphological evidence for tissue degeneration. The protein deposition was determined by immunohistochemistry on 30 complete cross-sections of lumbar spine obtained at autopsy (0-86 years) and 12 surgically removed disc samples. The mRNA expression was detected by non-radioactive in situ hybridization in the surgical material. All control experiments (blank and isotype controls in immunohistochemistry/sense controls in in situ hybridization) were negative. Immunohistochemically, we detected enhanced staining for fibronectin in both nuclear and anular tissues in areas with histological signs of mild-to-severe tissue degeneration (e.g., cleft formation and cell clustering) beginning with 16 years of age. Anular tissue showed less fibronectin staining than did nuclear areas. Fibronectin mRNA was detected mainly in nuclear cells by in situ hybridization corresponding to the protein staining indicating de novo synthesis. In parallel, TGF-beta1 was expressed by nuclear and occasional anular cells spatially associated with the fibronectin synthesizing cells. This was seen by both immunohistochemistry and in situ hybridization. This preliminary study provides evidence for a significant ongoing rearrangement of the extracellular matrix during disc degeneration, as monitored by enhanced fibronectin deposition that is produced by local disc cells. These cells also synthesize TGF-beta1, as shown by protein and mRNA expression. Since it is known that TGF-beta1 induces matrix alterations (by auto and paracrine stimulation of matrix synthesis), these observations suggest that the recently described disturbance of the matrix during disc degeneration may be induced by TGF-beta. This may offer new approaches to interfere with disc matrix alterations. PMID- 15340827 TI - Titanium mesh cages (TMC) in spine surgery. AB - The introduction of the titanium mesh cage (TMC) in spinal surgery has opened up a variety of applications that are realizable as a result of the versatility of the implant. Differing applications of TMCs in the whole spine are described in a series of 150 patients. Replacement and reinforcement of the anterior column represent the classic use of cylindrical TMCs. The TMC as a multisegmental concave support in kyphotic deformities and as a posterior interlaminar spacer or lamina replacement after wide laminectomy are additional applications. Implant subsidence, pseudarthrosis and implant loosening are the complications typically encountered with use of TMCs. The versatility of the implant permits its use in unusual surgical situations. PMID- 15340828 TI - Masting by Betula-species; applying the resource budget model to north European data sets. AB - Masting, the intermittent production of large crops of flowers by a plant population, is a common feature among trees in boreal and temperate forests. The pollen of many broadleaved trees causes allergic diseases, which are major causes of increasing health-care costs in industrialised countries. As the prevalence and severity of allergic diseases are connected with the concentrations of airborne pollen, an universal model predicting the intensity of the coming flowering would be a valuable tool for pollen information services, and ultimately for allergic people and allergologists. We investigated whether a resource budget model created in Japan explains the fluctuations in the annual pollen sums of Betula-species in north European data sets (10-12 years at 4 sites, 20 years at 10 sites). Using the shorter data sets, the model explained 76 92% of the annual fluctuations at five study sites. Using the 20-year data set, the percentage for southern Finland was much lower, only 48%, compared with the 85% of the 12-year data set. The annual pollen sums have been higher during the 1990s than in the 1980s, which may explain the ineffectiveness of the model, while applied to the 20-year data set. Our results support the resource budget model: the masting of birch species is regulated by weather factors together with the system of resource allocation among years. The model can serve pollen information service. However, only the 10 most recent years should be used to avoid interference from trends in changing vegetation and/or climate. PMID- 15340829 TI - Temporal variability in (13)C of respired CO(2) in a pine and a hardwood forest subject to similar climatic conditions. AB - Temporal variability in the (13)C of foliage (delta(13)C(F)), soil (delta(13)C(S)) and ecosystem (delta(13)C(R)) respired CO(2) was contrasted between a 17.2-m tall evenly aged loblolly pine forest and a 35-m tall unevenly aged mature second growth mixed broadleaf deciduous forest in North Carolina, USA, over a 2-year period. The two forests are located at the Duke Forest within a kilometer of each other and are subject to identical climate and have similar soil types. The delta(13)C(F), collected just prior to dawn, was primarily controlled by the time-lagged vapor pressure deficit (VPD) in both stands; it was used for calculating the ratio of intercellular to ambient CO(2) ( Ci/ Ca). A remarkable similarity was observed in the relationship between Ci/ Ca and time lagged VPD in these two forests despite large differences in hydraulic characteristics. This similarity emerged as a result of physiological adjustments that compensated for differences in plant hydraulic characteristics, as predicted by a recently proposed equilibrium hypothesis, and has implications to ecophysiological models. We found that in the broadleaf forest, the delta(13)C of forest floor CO(2) efflux dominated the delta(13)C(R), while in the younger pine forest, the delta(13)C of foliage respired CO(2) dominated delta(13)C(R). This dependence resulted in a more variable delta(13)C(R) in the pine forest when compared to the broadleaf forest due to the larger photosynthetic contribution. Given the sensitivity of the atmospheric inversion models to delta(13)C(R), the results demonstrate that these models could be improved by accounting for stand characteristics, in addition to previously recognized effects of moisture availability, when estimating delta(13)C(R). PMID- 15340830 TI - The dopaminergic nigrostriatal system: development, physiology, disease. PMID- 15340832 TI - Midbrain dopaminergic neurons: control of their cell fate by the engrailed transcription factors. AB - As for any other cell population, the development, cell fate, and properties of mesencephalic dopaminergic (mesDA) neurons are ultimately controlled at the transcriptional level. The genes for two transcription factors Engrailed-1 ( En1) and Engrailed-2 ( En2) play an essential role in the development and maintenance of these cells. They belong to a family of genes that have been investigated in Drosophila for more than half a century. The products of these genes are all characterized by homeotic tissue transformation and a highly conserved protein sequence, the homeobox. En1 and En2 act upon at least two steps of the differentiation of mesDA neurons. They take part in the regionalization event, which gives rise to the neuroepithelium that provides the precursor cells in the ventral midbrain with the fibroblast growth factor 8 signal necessary for their induction. Additionally, these genes are required in postmitotic mesDA neurons in which they are expressed from embryonic day 12 continuously into adulthood. In mutant mice homozygous null for En1 and En2, the neurons are generated in the ventral midbrain, become postmitotic, and begin to express their neurotransmitter phenotype. However, thereafter, they rapidly die by apoptosis. Cell mixing experiments in vitro and in vivo have demonstrated that the engrailed requirement for the survival of mesDA neurons is cell-autonomous. The inactivation of engrailed by RNA interference induces apoptosis in less than 24 h. These data suggest that the engrailed genes control an essential mechanism for the survival of mesDA neurons. PMID- 15340833 TI - Nurr1, an orphan nuclear receptor with essential functions in developing dopamine cells. AB - Nurr1 is a transcription factor that is expressed in the embryonic ventral midbrain and is critical for the development of dopamine (DA) neurons. It belongs to the conserved family of nuclear receptors but lacks an identified ligand and is therefore referred to as an orphan receptor. Recent structural studies have indicated that Nurr1 belongs to a class of ligand-independent nuclear receptors that are unable to bind cognate ligands. However, Nurr1 can promote signaling via its heterodimerization partner, the retinoid X receptor (RXR). RXR ligands can promote the survival of DA neurons via a process that depends on Nurr1-RXR heterodimers. In developing DA cells, Nurr1 is required for the expression of several genes important for DA synthesis and function. However, Nurr1 is probably also important for the maintenance of adult DA neurons and plays additional less well-elucidated roles in other regions of the central nervous system and in peripheral tissues. PMID- 15340834 TI - Insights into the western Bantu dispersal: mtDNA lineage analysis in Angola. AB - Africa is the homeland of humankind and it is known to harbour the highest levels of human genetic diversity. However, many continental regions, especially in the sub-Saharan side, still remain largely uncharacterized (i.e. southwest and central Africa). Here, we examine the mitochondrial DNA (mtDNA) variation in a sample from Angola. The two mtDNA hypervariable segments as well as the 9-bp tandem repeat on the COII/tRNA(lys) intergenic region have allowed us to allocate mtDNAs to common African haplogroups. Angola lies in the southern end of the putative western branch of the Bantu expansion, where it met the local Khoisan populations. Angolan mtDNA lineages show basically a Bantu substrate with no traces of Khoisan lineages. Roughly, more than half of the southwestern mtDNA pool can be assigned to west Africa, approximately 25% to central Africa and a significant 16% to east Africa, which points to the western gene pool having contributed most to the mtDNA lineages in Angola. We have also detected signals of extensive gene flow from southeast Africa. Our results suggest that eastern and western Bantu expansion routes were not independent from each other, and were connected south of the rainforest and along the southern African savannah. In agreement with historical documentation, the analysis also showed that the Angola mtDNA genetic pool shows affinities with the African lineages from Brazil, the main American destination of the slaves from Angola, although not all lineages in Brazil can be accounted for by the Angolan mtDNA pool. PMID- 15340835 TI - A large MSH2 Alu insertion mutation causes HNPCC in a German kindred. AB - Hereditary non-polyposis colorectal cancer (HNPCC) syndrome is an autosomal, dominantly inherited disease accounting for about 1%-5% of all colorectal cancer cases. HNPCC predisposition is caused by germline mutations in at least five genes coding for DNA mismatch repair (MMR) proteins. More than 400 MMR gene mutations have been identified in HNPCC patients. About 90% of mutations affect the MLH1 and MSH2 genes. The mutational spectrum mainly includes point mutations and small deletions or insertions. Here, we report a large 184 base-pair Alu insertion mutation in exon 6 of the MSH2 gene in a German HNPCC family. The inserted sequence contains repetitive Alu sequence elements that present the highest homology with the old Alu J subfamily. The Alu J insertion was most likely derived from Alu-mediated recombination, since Alu J elements have been found close to the insertion site in adjacent introns, and since elements pivotal for Alu retrotransposition are missing. Our results suggest that the recombination event occurred at least one generation ago. This is the first report of an Alu insertion in the coding sequence of a MMR gene as the cause of HNPCC. Our data thus further extend the spectrum of MMR gene mutations causative for HNPCC. PMID- 15340836 TI - Comparative mapping between Medicago sativa and Pisum sativum. AB - Comparative genome analysis has been performed between alfalfa (Medicago sativa) and pea (Pisum sativum), species which represent two closely related tribes of the subfamily Papilionoideae with different basic chromosome numbers. The positions of genes on the most recent linkage map of diploid alfalfa were compared to those of homologous loci on the combined genetic map of pea to analyze the degree of co-linearity between their linkage groups. In addition to using unique genes, analysis of the map positions of multicopy (homologous) genes identified syntenic homologs (characterized by similar positions on the maps) and pinpointed the positions of non-syntenic homologs. The comparison revealed extensive conservation of gene order between alfalfa and pea. However, genetic rearrangements (due to breakage and reunion) were localized which can account for the difference in chromosome number (8 for alfalfa and 7 for pea). Based on these genetic events and our increasing knowledge of the genomic structure of pea, it was concluded that the difference in genome size between the two species (the pea genome is 5- to 10-fold larger than that of alfalfa) is not a consequence of genome duplication in pea. The high degree of synteny observed between pea and Medicago loci makes further map-based cloning of pea genes based on the genome resources now available for M. truncatula a promising strategy. PMID- 15340837 TI - The cytoplasmic male-sterile type and normal type mitochondrial genomes of sugar beet share the same complement of genes of known function but differ in the content of expressed ORFs. AB - The complete nucleotide sequence (501,020 bp) of the mitochondrial genome from cytoplasmic male-sterile (CMS) sugar beet was determined. This enabled us to compare the sequence with that previously published for the mitochondrial genome of normal, male-fertile sugar beet. The comparison revealed that the two genomes have the same complement of genes of known function. The rRNA and tRNA genes encoded in the CMS mitochondrial genome share 100% sequence identity with their respective counterparts in the normal genome. We found a total of 24 single nucleotide substitutions in 11 protein genes encoded by the CMS mitochondrial genome. However, none of these seems to be responsible for male sterility. In addition, several other ORFs were found to be actively transcribed in sugar beet mitochondria. Among these, Norf246 was observed to be present in the normal mitochondrial genome but absent from the CMS genome. However, it seems unlikely that the loss of Norf246 is causally related to the expression of CMS, because previous studies on mitochondrial translation products failed to detect the product of this ORF. Conversely, the CMS genome contains four transcribed ORFs (Satp6presequence, Scox2-2 , Sorf324 and Sorf119) which are missing from the normal genome. These ORFs, which are potential candidates for CMS genes, were shown to be generated by mitochondrial genome rearrangements. PMID- 15340838 TI - PCR diagnosis of Pneumocystis carinii on sputum and bronchoalveolar lavage samples in immuno-compromised patients. AB - The polymerase chain reaction (PCR) technique was compared with Wright-Giemsa (WG), Gomori methenamine silver (GMS) stains and an immunofluorescence assay (IFA) for detection of Pneumocystis carinii in immuno-compromised patients. Specimens of 21 bronchoalveolar lavages (BAL) and 139 sputum samples, were obtained from 157 patients (38 with AIDS and 119 with HIV) from four hospitals in Khon Kaen, Thailand. A true positive required at least two positives by techniques considered gold standard tests. Eleven (52.38%) BAL and 13 (9.35%) sputum specimens were positive. PCR produced the highest sensitivity and negative predictive values for the BAL (100% for each) vs. sputum samples at 84.62 and 98.41 percent, respectively. The specificity of PCR was 90% and 98.41% for BAL and sputum samples, respectively. We suggest PCR is an important tool for the epidemiological study of P. carinii in high-risk individuals. PMID- 15340839 TI - Molecular phylogeny of isolates of Ctenocephalides felis and related species based on analysis of ITS1, ITS2 and mitochondrial 16S rDNA sequences and random binding primers. AB - The phylogenetic relationships among 31 different flea isolates representing seven different species were studied by nucleotide sequence comparison of the internal transcribed spacer 1 (ITS1), internal transcribed spacer 2 (ITS2) and/or mitochondrial 16S ribosomal RNA gene (mt16S-rDNA) to examine the patterns of variation. Results show that all regions are useful in discriminating among flea species. In Ctenocephalides felis and Tunga penetrans, some differences in these gene regions occurred among different isolates within the same species. In the latter case, the differences are in the mt16S-rDNA region, with one isolate showing 48% divergence in nucleotide sequence. The taxonomic implications of this result are unclear at present. The gene regions revealed differences between C. felis isolates only after DNA sequencing the PCR products. Further differentiation among C. felis isolates was obtained using four different random binding primers (decamers) and primers for mammalian aldolase to amplify narrow differences in the genome. Using these primers we were able to discriminate between different C. felis isolates and determine that some of the genetic variation coincided with minor differences in response to the control agent imidacloprid. However, overall findings do not support the existence of subspecies of C. felis. PMID- 15340840 TI - Molecular mechanisms of indirubin and its derivatives: novel anticancer molecules with their origin in traditional Chinese phytomedicine. AB - Indirubin, a 3, 2' bisindole isomer of indigo, has originally been identified as the active principle of a traditional Chinese preparation and has been proven to exhibit antileukemic effectiveness in chronic myelocytic leukemia. Indirubin was detected to represent a novel lead structure with potent inhibitory potential towards cyclin-dependent kinases (CDKs) resulting from high affinity binding into the enzymes ATP binding site. This seminal finding triggered our research to improve the pharmacological activities of the parent molecule within comprehensive structure-activity studies. Molecular modifications made novel anticancer compounds accessible with strongly improved CDK inhibitory potential and with broad spectrum antitumour activity. This novel family of compounds holds strong promise for clinical anticancer activity and might be useful also in several important noncancer indications, including Alzheimer's disease or diabetes. PMID- 15340841 TI - Reduced levels of the adenomatous polyposis coli (APC) protein are associated with ceramide-induced apoptosis of colon cancer cells. AB - PURPOSE: Mutations of the adenomatous polyposis coli (APC) and p53 genes are commonly found in colorectal cancers. We therefore analyzed the relative roles of APC and p53 in the induction of apoptosis of colon cancer cells by comparing the effects of the natural chemopreventive agent, C(2)-ceramide, on different human colon cancer cell lines with and without wild-type p53 and APC genes. METHODS: We studied the effect of C(2)-ceramide and C(2)-dihydroceramide on proliferation and/or apoptosis of colon cancer cell lines in vitro and determined the role of p53 and APC proteins in these processes. The protein and mRNA levels in colon cancer cell lines with and without treatments were determined by Western and Northern blot analysis, respectively. The cell cycle and apoptosis profiles were determined by FACS analysis and PARP-1 cleavage. RESULTS: Our findings indicate that C(2)-ceramide can induce apoptosis independently of the p53/p21(Waf-1/Cip-1) pathway. In addition, the C(2)-ceramide induction of apoptosis showed a correlation with a reduction in the levels of the APC protein and mRNA. Moreover, the C(2)-ceramide-induced apoptosis was blocked by pre-treatment with ZnCl(2), which stabilizes APC protein levels. CONCLUSIONS: These results suggest that C(2) ceramide treatment reduces the levels of APC protein and that the reduction in the levels of this protein plays a key role in the ability of C(2)-ceramide to induce apoptosis of colon cancer cells. PMID- 15340842 TI - KIAA0008 gene is associated with invasive phenotype of human hepatocellular carcinoma--a functional analysis. AB - PURPOSE: To investigate the function of the KIAA0008 gene, one of the leading genes in the signature associated with hepatocellular carcinoma (HCC) metastasis selected by cDNA microarray, and especially its possible roles in invasion and metastasis of hepatocellular carcinoma. METHODS: Expression levels of KIAA0008 in 27 primary tumors and 23 matched non-tumor liver tissues from HCC patients, and four HCC cell lines with different metastatic potentials were detected by semi quantitative RT-PCR and real-time RT-PCR. Recombinant expression plasmid vectors of the KIAA0008 gene were constructed and transfected into HCC cells. The subcellular localization of the KIAA0008 gene product and in vitro effects of KIAA0008 overexpression on proliferation and invasion of HCC cell line were also investigated. RESULTS: Expression levels of KIAA0008 in HCC tissues were statistically higher than those of paired non-tumorous liver tissues (P < 0.001, paired Wilcoxon test), and in HCCs with high invasiveness these were statistically higher than those with low invasiveness (P = 0.002, Mann-Whitney test). In the four HCC cell lines with an identical genetic background and stepwise higher invasiveness potentials, its expression was consistent with their invasiveness potential. The KIAA0008 gene product was concentrated on the nucleus and cell membrane of HCC cells, without any distribution in the cytoplasm. Overexpression of KIAA0008 in the MHCC97L cell line resulted in increased cell proliferation, colony formation, and invasion. CONCLUSIONS: KIAA0008 expression is associated with invasiveness of HCC; overexpression of KIAA0008 leads to a more invasive phenotype of HCC cell lines. PMID- 15340843 TI - Signal transduction of c-Kit receptor tyrosine kinase in CHRF myeloid leukemia cells. AB - PURPOSE: The tyrosine kinase receptor c-Kit (stem cell factor receptor, CD117) is a potential target for signal transduction therapy in different cancers. In this study we investigated c-Kit in CHRF cells, a megakaryoblastic cell line of Acute Myeloid Leukemia (FAB M7). MATERIALS AND METHODS: We characterized the interactions between c-Kit and PI 3-kinase (p85) after stimulation with SCF (stem cell factor) as well as the regulation of SHP-1 and SHP-2 associated with Kit in this cell line. RESULTS: Stimulation with SCF leads to a significant increase in interaction between Kit and p85 as well as in receptor associated PI 3-kinase activity. Interestingly, using different kinds of substances (AG 1295, CGP 53716) to inhibit the tyrosine kinase activity of c-Kit blocked activation of c-Kit, but the association of p85 still increased after SCF stimulation even when the tyrosine kinase activity of the receptor was completely blocked. In contrast, the other known interaction partners of c-Kit, SHP-1 and SHP-2, exhibited a basal association with c-Kit and no change of the association could be detected after stimulation of CHRF cells with SCF or treatment with the kinase inhibitors. CONCLUSIONS: Therefore, we suggest that association of p85, SHP-1, and SHP-2 to c Kit in CHRF cells can, at least in part, occur in a c-Kit kinase-activity independent manner. In contrast, the kinase activity of c-Kit is necessary for the activation of receptor-associated PI 3-kinase. PMID- 15340844 TI - Musculature of the crus and pes of the African elephant (Loxodonta africana): insight into semiplantigrade limb architecture. AB - The limbs of elephants are designed to support the weight of the largest terrestrial animal, and they display unique morphological peculiarities among mammals. In this article we provide a new and detailed anatomical description of the muscles of the lower hindlimb in African elephants (Loxodonta africana), and we place our observations into a comparative anatomical as well as a functional morphological context. At the cranial aspect of the shank (crus) and the foot (pes), the flexors of the tarsal joint and the extensors of the toes form a flat muscular plate covering the skeletal elements. Caudal to the tibia and the fibula the Musculus (M.) soleus is strongly developed, whereas the M. gastrocnemius and the M. flexor digitorum superficialis are thin. Small flexors, adductors, and abductors of the toes are present. The M. tibialis caudalis as well as the Mm. fibularis longus and brevis mainly support the tarsal joint. The design of the muscular structures matches the specific requirements of heavy-weight bearing as well as of proboscidean limb posture and locomotion patterns. PMID- 15340845 TI - Immunofluorescence mapping of dystrophin in the rat brain: astrocytes contain the splice variant Dp71f, but this is confined to subpopulations. AB - Dystrophins are membrane-associated actin-binding proteins, recognized at first in muscular dystrophies. In the brain the full-length Dp427 has been detected, as well as Dp140 and Dp71 of the shorter variants. Dp71 seems to be their major representative in the brain, and it occurs as splice variants, Dp71f and Dp71d. Dystrophins have been demonstrated mainly in neurons. In tissue cultures, the glial data, mainly in situ, are still insufficient. The present mapping study reveals the astroglial localization of the splice variant Dp71f, using a monoclonal antibody (5F3, developed by D. Mornet) specific for its additional 31 last amino acids. In parallel, another monoclonal antibody was used (Dys2, Novocastra) that detects the Dp71d, Dp427, as well as Dp140 and other short variants. Rats were overdosed with ether and perfused transcardially with 4% phosphate-buffered paraformaldehyde solution. Floating Vibratome sections were processed for immunohistochemical labeling with fluorescent secondary antibodies. In some animals the reactive glia were investigated following stab wounds in ketamine-xylazine anesthesia. Only the 5F3 antibody labeled astrocytes, however, not in general but in special localizations, mainly along the glia limitans of the pial surface, below the ependyma and in the reactive glia. Perivascular astrocytes were consistently labeled only where the vessels entered the brain, and in some circumventricular organs. The 5F3 antibody also labeled the ependyma and the residual subventricular zone. In contrast to the astrocytes, neurons were labeled throughout the brain. Dys2 antibody (to Dp71d and longer isoforms) labeled neurons in a distribution similar to that of 5F3, but rarely labeled astroglia and only in perivascular rings. Dp71f positivity seems to occur in those astrocyte populations that proved to be immunopositive to glial fibrillary acidic protein (GFAP) and produced laminin in former studies. PMID- 15340846 TI - Regulation of external genitalia development by concerted actions of FGF ligands and FGF receptors. AB - Members of the fibroblast growth factor (FGF) family play diverse roles during the development and patterning of various organs. In human and mice, 22 FGFs and four receptors derived from several splice variants are present. Redundant expression and function of FGF genes in organogenesis have been reported, but their roles in embryonic external genitalia, genital tubercle (GT), development have not been studied in detail. To address the role of FGF during external genitalia development, we have analyzed the expression of FGF genes (Fgf8, 9, 10) and receptor genes (Fgfr1, r2IIIb, r2IIIc) in GT of mice. Furthermore, Fgf10 and Fgfr2IIIb mutant mice were analyzed to elucidate their roles in embryonic external genitalia development. Fgfr2IIIb was expressed in urethral plate epithelium during GT development. Fgfr2IIIb mutant mice display urethral dysmorphogenesis. Marker gene analysis for urethral plate and bilateral mesenchymal formation suggests the existence of epithelial-mesenchymal interaction during urethral morphogenesis. Therefore, FGF10/FGFR2IIIb signals seem to constitute a developmental cascade for such morphogenesis. PMID- 15340848 TI - The time course of intended and unintended allocation of attention. AB - According to the contingent involuntary orienting hypothesis, only stimuli that match the attentional control settings based on intentions capture attention. In contrast, the surprise-capture hypothesis states that expectancy-discrepant stimuli can capture attention even if they do not match the control settings, implying unintended capture. The purpose of this study is to investigate whether unintended and intended attentional shifts are characterized by different time courses, indicating different underlying mechanisms. An unintended attentional shift was tested by the first, unannounced presentation of a color singleton at the location of a visual search target, and intended shifts by the following repeated presentations of a predictive singleton. Differences in time course were revealed by varying the stimulus onset asynchrony (SOA) between singleton and target. Results showed that accuracy with expected singletons was barely affected by SOA, whereas SOA strongly affected accuracy with the unexpected singleton. The results are interpreted as supporting the surprise-capture hypothesis. It is furthermore argued that a division of labor between contingent capture and surprise in the control of attention supports adaptive behavior. PMID- 15340847 TI - Cyclooxygenase-2 and its role in colorectal cancer development. AB - Cyclooxygenase 2 (COX-2), also called prostaglandin endoperoxide synthase 2, is involved in colorectal tumor development. This review deals with particular questions raised in this field such as the mechanisms of COX-2 related tumor promotion, the role of the different types of cells (epithelial and interstitial) expressing COX-2, the factors that trigger COX-2 induction, and the clinical potential of selective COX-2 inhibitors to treat or prevent colorectal tumors. Several mechanisms of COX-2 related tumor promotion have been identified. Some are dependent on prostaglandin E(2) production (such as induction of cell proliferation, angiogenenis or local immunosuppression, inhibition of apoptosis, increase in cell motility) and others are not (such as carcinogen activation or malondialdehyde production). COX-2 expression has been demonstrated in epithelial cells of colorectal cancers and adenomas and also in interstitial cells. These cells correspond to macrophages and/or fibroblasts and endothelial cells. COX-2 expression in these interstitial cells participates in tumor development. Factors or events that trigger COX-2 expression include oncogene activation, antioncogene inactivation, cytokines, growth factors, some fatty acids, bile salts, and mucins. Finally, selective COX-2 inhibitors may be effective in preventing or treating colorectal adenomas or carcinomas. However, their real efficiency and the cost/benefit balance are currently evaluated, and no definite conclusion can be made at the moment. PMID- 15340849 TI - Calcium-dependent changes in potassium currents in guinea-pig coronary artery smooth muscle cells after acute cobalt loading in vivo. AB - The aim of the present study was to determine whether cobalt poisoning induces haem oxidase isoenzyme-1 (HO-1) in coronary artery smooth muscle, or accounts for any changes in coronary smooth muscle cell (SMCs) membrane ionic currents that could result from this type of heavy metal poisoning. In SMCs isolated from cobalt-treated guinea-pig coronaries, K+ channel currents (IK) were much smaller than those in cells isolated from non-treated animals. Haemin (HO substrate) increased IK concentration dependently. This effect was mimicked by 1% CO and was abolished by pretreatment of cells with a competitive HO inhibitor, by inhibitors of guanylyl cyclase, protein kinase G or phospholipase C, as well as by blocking inositol trisphosphate-dependent Ca release, or sarcoplasmic reticulum Ca-ATPase, or by bathing cells in Ca-free external solution. Expression of the Na/Ca exchanger-1 (NCX-1) protein was reduced substantially in SMCs from coronary arteries of cobalt-treated animals. No expression of HO-1 was detected. It is concluded that acute cobalt poisoning in vivo depresses Ca-sensitive K currents via CO-dependent modulation of intracellular calcium availability, most probably by suppressing the expression of NCX-1 protein. PMID- 15340850 TI - Efflux properties of basolateral peptide transporter in human intestinal cell line Caco-2. AB - Small peptides and some pharmacologically active compounds are absorbed from the small intestine by the apical H(+)-coupled peptide transporter 1 (PEPT1) and the basolateral peptide transporter. Here we investigated the efflux properties of the basolateral peptide transporter in Caco-2 cells using two strategies, efflux measurements and a kinetic analysis of transepithelial transport of glycylsarcosine (Gly-Sar). [(14)C]Gly-Sar efflux through the basolateral membrane was not affected significantly by the external pH. Both approaches revealed that the basolateral peptide transporter was saturable in the efflux direction, and that the affinity was lower than that in the influx direction. For two peptide like drugs, there was no difference in substrate recognition by the basolateral peptide transporter between the two sides of the membrane. Using the kinetic parameters of PEPT1 and the basolateral peptide transporter, a computational model of Gly-Sar transport in Caco-2 cells was constructed. The simulation fitted the experimental data well. Our findings suggested that substrate affinity of the basolateral peptide transporter is apparently asymmetric, but pH-dependence and substrate specificity are symmetric for the two directions of transport. The behaviour of Gly-Sar in Caco-2 cells could be predicted by a mathematical model describing the peptide transporters. PMID- 15340852 TI - Emergence of orientation-selective inhibition in the primary visual cortex: a Bayes-Markov computational model. AB - The recent consensus is that virtually all aspects of response selectivity exhibited by the primary visual cortex are either created or sharpened by cortical inhibitory interneurons. Experimental studies have shown that there are cortical inhibitory cells that are driven by geniculate cells and that, like their cortical excitatory counterparts, are orientation selective, though less sharply tuned. The main goal of this article is to demonstrate how orientation selective inhibition might be created by the circuitry of the primary visual cortex (striate cortex, V1) from its nonoriented geniculate inputs. To fulfill this goal, first, a Bayes-Markov computational model is developed for the V1 area dedicated to foveal vision. The developed model consists of three parts: (i) a two-layered hierarchical Markov random field that is assumed to generate the activity patterns of the geniculate and cortical inhibitory cells, (ii) a Bayesian computational goal that is formulated based on the maximum a posteriori (MAP) estimation principle, and (iii) an iterative, deterministic, parallel algorithm that leads the cortical circuitry to achieve its assigned computational goal. The developed model is not fully LGN driven and it is not implementable by the neural machinery of V1. The model, then, is transformed into a fully LGN driven and physiologically plausible form. Computer simulation is used to demonstrate the performance of the developed models. PMID- 15340851 TI - Heat shock protein 70 inhibits shrinkage-induced programmed cell death via mechanisms independent of effects on cell volume-regulatory membrane transport proteins. AB - Cell shrinkage is a ubiquitous feature of programmed cell death (PCD), but whether it is an obligatory signalling event in PCD is unclear. Heat shock protein 70 (Hsp70) potently counteracts PCD in many cells, by mechanisms that are incompletely understood. In the present investigation, we found that severe hypertonic stress greatly diminished the viability of murine fibrosarcoma cells (WEHI-902) and immortalized murine embryonic fibroblasts (iMEFs). This effect was attenuated markedly by Hsp70 over-expression. To determine whether the protective effect of Hsp70 was mediated via an effect on volume regulatory ion transport, we compared regulatory volume decrease (RVD) and increase (RVI) in control WEHI-902 cells and after increasing Hsp70 levels by heat shock or over-expression (WEHI 912). Hsp70 levels affected neither RVD, RVI nor the relative contributions of the Na(+)/H(+)-exchanger (NHE1) and Na(+),K(+),2Cl(-)-cotransporter (NKCC1) to RVI. Hypertonic stress induced caspase-3 activity in WEHI cells and iMEFs, an effect potentiated by Hsp70 in WEHI cells but inhibited by Hsp70 in iMEFs. Osmotic shrinkage-induced PCD was associated with Hsp70-inhibitable cysteine cathepsin release in iMEFs and attenuated by caspase and cathepsin inhibitors in WEHI cells. Treatment with TNF-alpha or the NHE1 inhibitor 5'-(N-ethyl-N isopropyl)amiloride (EIPA) reduced the viability of WEHI cells further under isotonic and mildly, but not severely, hypertonic conditions. Thus, it is concluded that shrinkage-induced PCD involves both caspase- and cathepsin dependent death mechanisms and is potently counteracted by Hsp70. PMID- 15340853 TI - Active oxygen and nitrogen species in biology: from cytocidal agents to signaling intermediates. PMID- 15340854 TI - No effects of intermittent 50 Hz EMF on cytoplasmic free calcium and on the mitochondrial membrane potential in human diploid fibroblasts. AB - The recently described increase in DNA strand breaks of cultured human diploid fibroblasts after intermittent exposure to extremely-low-frequency electromagnetic fields (ELF-EMF) of more than about 70 microT ELF-EMF is difficult to explain by a direct induction of covalent bond disruption. Therefore the hypothesis has been tested that ELF-EMF-induced DNA strand breaks might be mediated by cellular processes that cause alteration of the intracellular concentration of free calcium ([Ca2+]i) and/or the membrane potential (DeltaPsi(m)). [Ca2+]i was determined by the ratiometric fura-2 technique. Changes in DeltaPsi(m) were assessed by using the potential-dependent lipophilic cationic probe JC-1. Human fibroblasts were exposed to intermittent ELF-EMF (50 Hz, 1000 microT). Although exposure of fiboblasts to ELF-EMF resulted in a highly significant increase in DNA strand breaks as determined by the comet assay, no effect on JC-1 fluorescence emission or on [Ca2+]i has been observed when comparing exposed with sham-exposed cells. Therefore, it is suggested that ELF EMF-induced DNA strand breaks are unlikely to be caused by intracellular changes that affect [Ca2+]i and/or DeltaPsi(m). PMID- 15340855 TI - Assignment of the 2.03 ppm resonance in in vivo 1H MRS of human brain tumour cystic fluid: contribution of macromolecules. AB - MRI and MRS are established techniques for the evaluation of intracranial mass lesions and cysts. The 2.03 ppm signal recorded in their (1)H-MRS spectra is often assigned to NAA from outer volume contamination, although it has also been detected in non-infiltrating tumours and large cysts. We have investigated the molecular origin of this resonance in ten samples of cystic fluids from human brain tumours. The NMR detected content of the 2.03 ppm resonance in 136 ms echo time spectra, assuming an N- CH(3) origin, was 3.19 +/- 1.01 mM. Only one third (34 +/- 12%) of the N-acetyl containing compound (NAC) signal could be extracted by perchloric acid (PCA) indicating that most of it originated in a macromolecular PCA-insoluble component. Chemical analysis of the cyst fluids showed that sialic acid bound to macromolecules would account for 64.3% and hexuronic containing compounds for 29.2% of the NMR-detectable ex vivo signal, 93.4% of the signal at TE 136 ms. Lactate content measured by NMR (6.4 +/- 4.4 mM) and the predominance of NAC originating in sialic acid point to a major origin from tumour rather than from plasma for this 2.03 ppm resonance. PMID- 15340856 TI - High cerebral scyllo-inositol: a new marker of brain metabolism disturbances induced by chronic alcoholism. AB - Cerebral metabolic changes that concur to motor and/or cognitive disorders in actively drinking alcoholics are not well established. We tested the hypothesis that chronic alcoholics exhibit profound alterations in the cerebral metabolism of scyllo-inositol. Brain metabolism was explored in nine actively drinking and 11 recently detoxified chronic alcoholics by in vivo brain (1)H-MRS and in vitro(1)H-MRS of blood serum and cerebrospinal fluid. The cohort was composed of individuals with acute, subacute or chronic encephalopathy or without any clinical encephalopathy. Chronic alcoholism is associated with a hitherto unrecognized accumulation of brain scyllo-inositol. Our results suggest that scyllo-inositol is produced within the central nervous system and shows a diffuse but heterogenous distribution in brain where it can persist several weeks after detoxification. Its highest levels were observed in subjects with a clinically symptomatic alcohol-related encephalopathy. When detected, brain scyllo-inositol takes part in a metabolic encephalopathy since it is associated with reduced N acetylaspartate and increased creatine. High levels of cerebral scyllo-inositol are correlated with altered glial and neuronal metabolism. Our findings suggest that the accumulation of scyllo-inositol may precede and take part in the development of symptomatic alcoholic metabolic encephalopathy. PMID- 15340857 TI - A new pulse sequence to visualize slow flow. AB - The purpose of this paper is to present a new pulse sequence for visualizing slow flow. The new sequence consists of an initial Stejskal-Tanner flow sensitization part followed by a DEFT pulse and a spoiler gradient. A single-shot TSE readout train is then applied to sample the NMR signal. The sequence was initially tested using a simple flow phantom. To verify potential clinical use, both flow sensitive MRCP and cerebrospinal fluid (CSF) images were produced. The phantom study proved the sequence sensitivity to flow in the range 0-1 cm/s. bVE-factors 1.5, 3, 6 and 12 were chosen. Within this flow velocity range, the signal dropped as predicted theoretically. This indicates that the method can be used to quantify flow. All anatomical features seen in a standard MRCP sequence were identified and the methods sensitivity to CSF flow was demonstrated by sagital images of the head. A new pulse sequence sensitive to slow flow has been developed. PMID- 15340858 TI - Does relative economic value of food elicit purposeful encounter in the yellow baboons (Papio hamadryas cynocephalus) of Ruaha National Park, Tanzania? AB - In the dry season, baboons traveled purposefully to spatially predictable foods that provided a relatively large number of grams per minute of preparation (e.g., economical foods), but not to predictable foods that merely accounted for a large proportion of feeding time (Pochron in Int J Primatol 22:773-785, 2001). In this paper, I examined the generality of those findings across seasons and applied the same methods to baboons eating a lush-season diet. I hypothesized that baboons should travel quickly and directly (i.e., purposefully) only to economical foods. The change in diet brought about by season provided an important comparison. In the lush season, none of the spatially predictable foods provided a relatively large number of grams per minute of preparation, so baboons were predicted to travel purposefully to no lush-season food. In short, baboons who traveled quickly and directly to some foods in the dry season were expected to use indirect and/or slow travel for all lush-season foods. The hypothesis was supported--baboons traveled quickly and directly to no lush-season food. Detailed comparisons between the dry- and lush-season characteristics of baobab fruit show that the food's economic value relative to other foods predicts concurrent fast and direct travel, or the lack thereof, in both seasons. PMID- 15340859 TI - On dense granular flows. AB - The behaviour of dense assemblies of dry grains submitted to continuous shear deformation has been the subject of many experiments and discrete particle simulations. This paper is a collective work carried out among the French research group Groupement de Recherche Milieux Divises (GDR MiDi). It proceeds from the collection of results on steady uniform granular flows obtained by different groups in six different geometries both in experiments and numerical works. The goal is to achieve a coherent presentation of the relevant quantities to be measured i.e. flowing thresholds, kinematic profiles, effective friction, etc. First, a quantitative comparison between data coming from different experiments in the same geometry identifies the robust features in each case. Second, a transverse analysis of the data across the different configurations, allows us to identify the relevant dimensionless parameters, the different flow regimes and to propose simple interpretations. The present work, more than a simple juxtaposition of results, demonstrates the richness of granular flows and underlines the open problem of defining a single rheology. PMID- 15340860 TI - Fluid membranes exchanging material with external reservoirs. AB - We investigate the dynamics of a single component fluid bilayer, which exchanges material with the surrounding fluid. We derive covariant equations of motion taking into account solvent permeation, exchange of lipids between solvent and the membrane and discuss the sources of noise in these equations. Different lipid concentrations on both sides of the membrane lead to a non-equilibrium state. We discuss steady states as well as shape instabilities which occur at a critical osmotic-pressure difference. PMID- 15340861 TI - Some remarks on the rheology of dense granular flows. AB - The paper reviews some peculiar properties exhibited by granular flows. We emphasize the inability of kinetic theory and of Bagnold's heuristic approach to describe the rapid regime of densely packed flows, characterized by the breakdown of the binary collision picture and by multibody long-lasting contacts. We suggest that deformation waves through the continuous paths of contacts can be effective to transport momentum and energy through the bulk, in a time very short compared to the inverse of the shear rate. This mechanism could explain some key rheological features encountered in dense granular materials. PMID- 15340862 TI - Influence of the spatial beam profile on hard tissue ablation, part II: pulse energy and energy density distribution in simple beams. AB - When calculating applied flux densities in practice, the beam profile of a laser is often erroneously assumed to be homogeneous. In addition, there is usually no consistency in the choice of a suitable measuring method for determining the beam diameter. This failure to observe the inhomogeneous intensity distribution within the beam cross-section, combined with the imprecise knowledge of the beam diameter, leads to flux densities being stated that represent mean values at best. The present paper gives definitions for the flux densities of simple, radially symmetrical beam cross-sections, taking the top-hat and Gaussian profiles as examples. In connection with the inhomogeneous energy distribution in the Gaussian beam, a concept of integral and local energy density is discussed, which differs from the customary definition of the energy density as a constant. Also presented are the consequences of the mathematical concepts in terms of measurement, giving particular consideration to the case where the energy density as the measured variable matches the integral energy density. The significance of the integral and local energy density for hard-tissue ablation is described, based on the practical example of the ablation of dental hard substance. The central result is that the integral flux density is directly accessible as a measured variable, while the effect on the tissue is determined by the local flux density. If the form of the beam is known, the integral flux density can be converted into the local flux density. PMID- 15340863 TI - 5-aminolevulinic acid ester-induced protoporphyrin IX in a murine melanoma cell line. AB - The use of 5-aminolevulinic acid (5-ALA) ester derivatives as precursors of endogenous protoporphyrin IX (PpIX) has been proposed as a good strategy for improved drug diffusion across biological membranes. In the present work, the 5 ALA ester derivatives hexyl-ALA (h-ALA), octyl-ALA (o-ALA), and decyl-ALA (d-ALA) were synthesized, and their efficacy to induce endogenous PpIX was explored in a murine melanoma cell line (B-16) as compared with that of 5-ALA. The maximum level of PpIX induced in cells treated with 5-ALA, h-ALA, o-ALA, and d-ALA was reached at optimal concentrations of 0.3, 0.075, 0.1, and 0.075 mM, respectively. The derivatives h-ALA and o-ALA appear as the most efficient PpIX precursors in this cell line, since a higher or similar PpIX production could be achieved with a fourfold and threefold lower dose of these precursors compared with 5-ALA. The phototoxicity effect of h-ALA and o-ALA ester derivatives showed the same phototoxicity behavior detected for 5-ALA but at much lower drug doses. Our study suggests that h-ALA and o-ALA esters improve intracellular PpIX formation in B-16 cells at reduced concentrations. This should enable clinical applications at lower precursor doses with reduced effective costs. PMID- 15340864 TI - Health-related quality of life in rheumatoid arthritis in Northern Sweden: a comparison between patients with early RA, patients with medium-term disease and controls, using SF-36. AB - Living with a chronic disease affects many aspects of an individual's life. The aim of this study was to compare the health-related quality of life, as measured by the SF-36, in patients with early rheumatoid arthritis (RA) at disease onset and after 2 years. The results were furthermore compared with those of patients with medium-term disease and a control group. Forty patients with early RA as well as 39 RA patients with 21-25 years of disease duration and 40 controls were asked to answer the self-administered SF-36 health profile measure. Both patients with early RA and with medium-term disease reported significantly lower values for all eight subscales compared with the controls. At follow-up after 2 years, the patients reported significant improvements on the role physical (RP) and bodily pain (BP) dimensions compared with at disease onset. Physical functioning (PF) was perceived as better in patients with early RA compared with patients who had had the disease for 21-25 years. Women reported significantly higher values for some of the scales than men. In summary, health-related quality of life is negatively affected in early RA as measured by the SF-36. An improvement was implicated after 2 years. There were some gender differences in reported health related quality of life among patients with early RA, but not in patients with medium-term disease. PMID- 15340865 TI - Familial inflammatory Sneddon's syndrome-case report and review of the literature. AB - Sneddon's syndrome (SNS) which originally was a clinical diagnosis, is now regarded as a common clinical manifestation of different disease entities. It has been divided into idiopathic, autoimmune and thromboembolic subsets or in systemic lupus erythematosus (SLE)-associated, antiphospholipid syndrome (APS) associated and primary forms. Familial occurrence of Sneddon's syndrome is rare. We present a familial case of Sneddon's syndrome with inflammatory disease pattern, early disease onset and association with autoimmune thyroid disease and anticardiolipin antibodies. Although most authors reporting on adult cases of SNS consider it a non-inflammatory, thromboembolic process, the study of cases with early onset brings attention to the possible inflammatory origin of the syndrome. PMID- 15340866 TI - The Turkish version of the Bath Ankylosing Spondylitis Functional Index: reliability and validity. AB - The purpose of this study was to investigate the reliability and validity of the Turkish version of the Bath Ankylosing Spondylitis (AS) Functional Index (BASFI). The Turkish version of the BASFI was obtained after a process of translation and back-translation. Eighty-one consecutive patients meeting the 1984 New York criteria for AS were enrolled. Patients were evaluated and requested to complete the questionnaire at days 1 and 2 and on a third occasion between days 15-90. Reliability, reproducibility, validity and sensitivity to change of the Turkish version of the index were assessed. Each score correlated closely with the index score, with coefficients between 0.727 and 0.844. Reliability analysis showed a Cronbach's alpha score of 0.926. Correlations were found between all items of the BASFI and Schober's test (r=-0.258 to -0.531, p<0.001-0.05), occiput-to-wall distance (r=0.284 and 0.589, p<0.001-0.05), and finger-to-floor distance (r=0.334 to 0.613, p<0.001-0.01). The total index score was correlated with the number of nocturnal awakenings (r=0.515, p<0.001), Schober's test (r=-0.444, p<0.001), finger-to-floor distance (r=0.567, p<0.001), occiput-to-wall distance (r=0.535, p<0.001), chest expansion (r=-0.403, p<0.001), and the Dougados articular index (r=0.371, p<0.01). A good correlation was found between day 0 and 1 BASFI indices (r=0.765-0.917, p<0.001), showing good reproducibility of the index. The Turkish version of the BASFI showed reliability, reproducibility, and validity, confirming its utility in the research of AS in Turkey. However, sensitivity to changes due to drug therapy and/or rehabilitation remains to be determined. PMID- 15340867 TI - Analysis of hydrostatic pressure effects on transcription in Escherichia coli by DNA microarray procedure. AB - Hydrostatic pressure is a well-known physical stimulus, but its effects on cell physiology have not been clarified. To investigate pressure effects on Escherichia coli, we carried out DNA microarray analysis of the entire E. coli genome. The microarray results showed pleiotropic effects on gene expression. In particular, heat- and cold-stress responses were induced simultaneously by the elevated pressure. Upon temperature stress (including both temperature up- and down-shifts) and other environmental stresses, gene expression adjusts to adapt to such environmental changes through regulations by several DNA-binding proteins. An E. coli mutant, which deleted the hns gene encoding one of the regulator proteins, exhibited great pressure sensitivity. The result suggested that the H-NS protein was a possible transcriptional regulator for adaptation of the high-pressure stress. PMID- 15340868 TI - Microperoxidase 8 adsorbed on a roughened silver electrode as a monomeric high spin penta-coordinated species: characterization by SERR spectroscopy and electrochemistry. AB - Microperoxidase 8 (MP8), a heme octapeptide obtained by hydrolytic digestion of cytochrome c, was adsorbed at the surface of a roughened silver electrode in order to provide a new supported biomimetic system for hemoproteins. A combination of two techniques was used to study its redox and coordination properties: electrochemistry and surface-enhanced resonance Raman (SERR) spectroscopy. This allowed us to show that MP8 could be adsorbed as a monolayer at the surface of the roughened silver electrode, where it could undergo a reversible electron transfer. Under those conditions, a redox potential of -0.4 V vs. SCE (-0.16 V vs. NHE) was measured for MP8, which was almost identical to that reported for N-acetyl-MP8 in aqueous solution. In addition, whereas MP8 appeared to aggregate in solution, and led to a mixture of high-spin penta coordinated (5cHS) and low-spin hexa-coordinated (6cLS) iron(III) or iron(II) species, it was recovered almost exclusively as a monomeric high-spin penta coordinated species at the surface of the electrode, both in the reduced and in the oxidized states. This then allowed a free coordination site on the iron, on the distal face of MP8 accessible to ligands. Accordingly, experiments performed in the presence of potassium cyanide demonstrated that MP8 adsorbed on a silver electrode could be ligated by a sixth CN(-) ligand. Thus there is the possibility of binding several kinds of ligands such as O(2) or H(2)O(2), which will open the way to biocatalysis of oxidation reactions at the surface of an electrode, or ligands such as drugs which will lead to the design of new biosensors for molecules of biological interest. PMID- 15340869 TI - Clinical advantages of COMT inhibition with entacapone - a review. AB - Two catechol- O-methyltransferase (COMT) inhibitors, entacapone and tolcapone, were developed during the 1990's to be used as adjuncts to levodopa (LD) - dopa decarboxylase (DDC) inhibitors in the treatment of Parkinson's disease (PD). Entacapone is currently in wide clinical use, while tolcapone can be used in restricted indications only, due to its hepatotoxicity. COMT inhibitors prolong the elimination of LD, while DDC inhibitors mainly increase its absorption; both mechanisms leading to increased bioavailability of LD. The pharmacokinetic properties of LD, carbidopa and entacapone are quite similar, and entacapone is administered concomitantly with LD plus carbidopa. Entacapone prolongs the clinical effect of each LD dose by 30 to 40 minutes; this effect is seen already after the first entacapone dose. When LD is administered in several frequent daily doses, addition of entacapone reduces the daily fluctuations of plasma LD by 30 to 40%. Based on studies with home diaries, entacapone increases the daily ON-time by an average of one to two hours, and reduces the daily OFF-time correspondingly in patients with PD with motor fluctuations. The daily LD dose has been reduced by 10 to 30%. These positive effects are sustained in long term use over several years. There is still scant information of the benefit of entacapone in patients without motor fluctuations. Entacapone can cause both dopaminergic and non-dopaminergic adverse events. Increased dyskinesias are most frequently recorded in patients with motor fluctuations. The dopaminergic adverse events can usually be diminished by reducing the LD dose. Non-dopaminergic adverse events are abdominal pain and diarrhea. Diarrhoea has led to discontinuation in 3 to 4% of the patients in clinical trials. Entacapone has not been connected to liver toxicity and there are no indications to follow laboratory safety during treatment. The benefit-risk ratio of entacapone is considered favorable.A triple LD/carbidopa/entacapone combination tablet has recently been developed. Three LD strengths (50, 100 and 150 mg) are available, each contains 200 mg of entacapone. The majority of the patients can be managed with these three LD strengths. Entacapone has today an established position in treatment of PD patients with motor fluctuations, either as a separate tablet or as the triple LD combination. PMID- 15340870 TI - Parkinsonism following striatal infarcts: incidence in a prospective stroke unit cohort. AB - A number of case reports have highlighted the occurrence of parkinsonism following strategic infarcts affecting the basal ganglia but the prevalence of parkinsonism after striatal infarcts (SI) has not been assessed. Therefore, we evaluated the clinical features and prevalence of parkinsonism in a large series of patients admitted to the Stroke-Unit of the Department of Neurology Innsbruck. Cerebral scans were retrospectively screened for SI, defined as a lesion larger than 1.5 cm involving the basal ganglia and the internal capsule. Out of 622 patients, 27 met the criteria for SI (4.3%) and 11 patients were available for follow-up. All patients presented contralateral motor weakness. Bilateral akinetic-rigid parkinsonism was found in only one patient whose [(123)I]beta-CIT SPECT showed a decrease of the ligand uptake following the limits of the vascular lesion. Overall, parkinsonism does not appear to be a frequent consequence of striatal infarcts. Multiple lacunar subcortical infarcts interrupting thalamocortical drive may be more critical for the development of vascular parkinsonism. PMID- 15340871 TI - Subsecond changes of global brain state in illusory multistable motion perception. AB - This study explored transient changes in EEG microstates and spatial Omega complexity associated with changes in multistable perception. 21-channel EEG was recorded from 13 healthy subjects viewing an alternating dot pattern that induced illusory motion with ambiguous direction. Baseline epochs with stable motion direction were compared to epochs immediately preceding stimuli that were perceived with changed motion direction ('reference stimuli'). About 750 ms before reference stimuli, Omega complexity decreased as compared to baseline, and two of four classes of EEG microstates changed their probability of occurrence. About 300 ms before reference stimuli, Omega complexity increased and the previous deviations of EEG microstates were reversed. Given earlier results on Omega complexity and microstates, these sub-second EEG changes might parallel longer-lasting fluctuations in vigilance. Assumedly, the discontinuities of illusory motion thus occur during sub-second dips in arousal, and the following reconstruction of the illusion coincides with a state of relative over-arousal. PMID- 15340872 TI - Re-evaluation of the dysfunction of mitochondrial respiratory chain in skeletal muscle of patients with Parkinson's disease. AB - The origin and tissue distribution of the mitochondrial dysfunction in Parkinson's disease (PD) remains still a matter of controversy. To re-evaluate a probably free radical-born, generalized mitochondrial impairment in PD, we applied optimized enzymatic assays, high resolution oxygraphic measurements of permeabilized muscle fibers, and application of metabolic control analysis to skeletal muscle samples of 19 PD patients and 36 age-matched controls. We detected decreased activities of respiratory chain complexes I and IV being accompanied by increased flux control coefficients of complexes I and IV on oxygen consumption of muscle fibers. We further investigated if randomly distributed point mutations in two discrete regions of the mitochondrial DNA are increased in PD muscle, and if they could contribute to the mitochondrial impairment. Our data confirm the previously debated presence of a mild mitochondrial defect in skeletal muscle of patients with PD which is accompanied with an about 1.5 to 2-fold increase of point mutated mtDNA. PMID- 15340873 TI - Ruptured aneurysm at a duplicated middle cerebral artery with accessory middle cerebral artery. AB - Although many cerebral vascular anomalies are widely recognized, others are less well known or unclassified. Accessory middle cerebral artery (MCA) and duplicated MCA are among uncommon anomalies. We present a very rare case of subarachnoid haemorrhage due to rupture of a saccular aneurysm arising from a duplicated middle cerebral artery which was associated with an accessory middle cerebral artery. PMID- 15340874 TI - The effects of ageing on brain tissue sialic acid contents following cold trauma. AB - BACKGROUND: There is a prominent difference between the responses of young and aged patients to brain injury in the clinical setting, but the exact cause of this condition is not well known. METHODS: Young (3-4 months) and aged (36-40 months) Wistar albino male rats were used as subjects, and they were divided into four groups: young and aged study groups, and young and aged control groups. In all groups, craniectomies were performed over the left hemispheres, and in the study groups, cold injuries were inflicted. Brain tissue sialic acid contents were determined in all groups. FINDINGS: Brain tissue sialic acid content was higher in aged control rats than young control ones, but the difference was not statistically significant. Tissue sialic acid content was significantly increased in young rats after trauma. On the contrary, it was significantly decreased in aged rats after trauma. INTERPRETATION: In young rats, brain tissue sialic acid content was significantly increased 24 hours after cold injury unlike in the aged ones. This may be a finding related to decreased regeneration capability of aged brain. PMID- 15340875 TI - Parameters influencing augmentation of cerebral blood flow by cervical spinal cord stimulation. AB - BACKGROUND: Cervical spinal cord stimulation (SCS) has been shown to augment cerebral blood flow (CBF) and protect the brain from focal ischemia. However, the application of SCS in the treatment of cerebral ischemia requires a better understanding of the limits of the cerebrovascular effect and the optimal stimulation parameters. In the present study, we investigated the effects of various stimulation parameters on CBF augmentation, as well as the issue of tachyphylaxis of the CBF response. METHODS: SCS was performed in adult Sprague Dawley rats, and CBF was assessed using cortical laser Doppler flowmetry (LDF). In separate experimental series, stimulation amplitude, frequency, and pulse width were varied, and the effect on the LDF response was recorded. Finally, using the stimulation parameters found to elicit the strongest LDF response, we examined the effect of lengthening the period of SCS. FINDINGS: SCS elicited a robust increase in cortical LDF values as previously demonstrated. The magnitude of the response varied in a dose-dependent fashion with the stimulation amplitude. LDF values increased by more than 80% over baseline with an amplitude of 1.5 mA. The optimal pulse width and frequency of the stimulation were 0.25 ms and 50 Hz, respectively. Lengthening the stimulation period up to 20 minutes resulted in a persistent increase in cerebral LDF values during the entire stimulation period, although the magnitude of this effect diminished to approximately 50% over the baseline after 10 minutes. CONCLUSIONS: SCS elicits a robust augmentation in CBF, which lasts the entire stimulation duration. Stimulation parameters required for optimal cerebrovascular response are within normally used therapeutic ranges in the clinical settings. These results provide further evidence that SCS may provide a novel therapeutic strategy for the treatment of cerebral ischemia. PMID- 15340876 TI - Dissecting aneurysm of the posterior communicating artery. AB - A 36-year-old male presented with an extremely rare dissecting aneurysm of the posterior communicating artery manifesting as severe occipital headache. Magnetic resonance (MR) imaging revealed a partially thrombosed aneurysm of the right posterior communicating artery and angiography showed the pearl and string sign. Three months later, repeat angiography showed that the aneurysm was completely thrombosed and the posterior communicating artery was occluded. Magnetic resonance (MR) imaging showed an intimal flap. These neuroradiological findings demonstrated that the aneurysm was a dissecting aneurysm of the right posterior communicating artery. He was discharged with no neurological deficit. PMID- 15340877 TI - Skull osteosarcoma: illustrated review. AB - A case of conventional intramedullary osteosarcoma (epithelioid subtype) with 10 years of evolution and another of high grade surface osteosarcoma of the chondroblastic type, both in the skull, gave rise to several diagnostic and therapeutic difficulties. Key points concerning the definition, classification, imaging, clinical series data and treatment options available for skull osteosarcoma are highlighted. PMID- 15340878 TI - WITHDRAWN: Microsurgical resection of intramedullary spinal cord metastases of large cell lung cancer. AB - This article has now been withdrawn and will not appear in print. PMID- 15340879 TI - Functional food: a growing and not clearly controlled market with a risk potential ranging from a food to a drug. PMID- 15340880 TI - The relationship between schizophrenia and religion and its implications for care. AB - This paper focuses on the relationships between schizophrenia and religion, on the basis of a review of literature and the data of an ongoing study about religiousness and spiritual coping conducted among outpatients with chronic schizophrenia. Religion (including both spirituality and religiousness) is salient in the lives of many people suffering from schizophrenia. However, psychiatric research rarely addresses religious issues. Religious beliefs and religious delusions lie on a continuum and vary across cultures. In Switzerland for example, the belief in demons as the cause of mental health problems is a common phenomenon in Christians with high saliency of religiousness. Religion has an impact, not always positive, on the comorbidity of substance abuse and suicidal attempts in schizophrenia. In many patients' life stories, religion plays a central role in the processes of reconstructing a sense of self and recovery. However religion may become part of the problem as well as part of the recovery. Some patients are helped by their faith community, uplifted by spiritual activities, comforted and strengthened by their beliefs. Other patients are rejected by their faith community, burdened by spiritual activities, disappointed and demoralized by their beliefs. Religion is relevant for the treatment of people with schizophrenia in that it may help to reduce pathology, to enhance coping and to foster recovery. In the treatment of these patients, it appears useful to tolerate diversity, to respect others beliefs, to ban proselytism and to have a good knowledge of one's own spiritual identity. PMID- 15340881 TI - Montelukast and Churg-Strauss syndrome. AB - The association of leukotriene receptor antagonists and Churg-Strauss Syndrome (CSS) has been recognised for several years. However, whether these drugs have a direct pathogenic role remains controversial. The present case describes an asthmatic patient, who developed severe obstructive symptoms and progressive heart failure after two sequential exposures to montelukast. As the patient exhibited a markedly raised blood eosinophil count with diffuse infiltrates on chest x-ray and signs of myocarditis, CSS was suspected. The disease was confirmed by open lung biopsy. The symptoms improved rapidly after administration of high dose immunosuppression with methylprednisolone and cyclophosphamide. This case is noteworthy because the time course of events strongly suggests a direct aetiological role for montelukast in the development of CSS. The pathophysiological mechanism of the association remains unknown. PMID- 15340882 TI - A diminished postnatal surge of Ad spermatogonia in cryptorchid infants is additional evidence for hypogonadotropic hypogonadism. AB - BACKGROUND: The aim of this study was to describe the development of Ad spermatogonia in both unilateral and bilateral cryptorchid infants compared to a control population of comparable age and to note particularly the fate of Ad spermatogonia during the normal surge of testosterone and gonadotropin. METHODS: The incidence and development of Ad (dark) adult type of spermatogonia were assessed in 270 testicular biopsies from 159 cryptorchid infants at 1-12 months of age. These results were compared to the control population of the same age. RESULTS: The number of Ad spermatogonia increased markedly after five months of life in the control population. The scrotal testes of unilateral cryptorchid infants also had an increase in the number of Ad spermatogonia but it was distinctly lower than that of the control population. In contrast, this surge was completely absent in the cryptorchid testes. The number of Ad spermatogonia in unilateral cryptorchid testes correlated in a nonlinear fashion with those in the contralateral scrotal testes. The total number of germ cells in the cryptorchid testes in the first six months of life is normal, after which time it declines rapidly. CONCLUSION: The impaired transformation of germ cells into Ad spermatogonia in both unilateral and bilateral cryptorchid infant testes during mini-puberty underscores the importance of hypogonadotropic-hypogonadism in the pathogenesis of this disease. PMID- 15340883 TI - Interaction of food supplement L-carnitine with oral anticoagulant acenocoumarol. PMID- 15340885 TI - [Validation of a screening questionnaire for stroke detection in Spanish-speaking communities]. AB - OBJECTIVE: To evaluate the accuracy of an screening questionnaire for stroke detection in speaking-speaking communities. PATIENTS AND METHODS: We performed a door-to-door survey in Atahualpa (a rural community in coastal Ecuador) using a modified version of the Spanish translation of the OMS questionnaire for stroke detection. Subjects suspected of having a stroke as well as a 2% sample of negative subjects were evaluated by neurologists to evaluate sensitivity, specificity, and predictive value of the questionnaire and that of each of its questions. RESULTS: We found 18 possible cases among 1,568 individuals around 15 years old. Of these, 10 were confirmed stroke patients and 8 were false-positive (crude stroke prevalence of 6.38 per 1,000). We did not find false-negative cases. Sensitivity of the questionnaire was 100%, specificity was 99.5%, positive predictive value was 0.55 and negative predictive value was 1. The accuracy of each question as well as the number of questions answered as affirmative were different between patients and false-positive cases. CONCLUSIONS: The current questionnaire is highly sensitive but its positive predictive value is poor. This causes problems in large-scale studies, as the detection of many false-positives may compromise its viability. We propose a modification of the questionnaire that will turn it more accurate. PMID- 15340886 TI - [Stroke and intracranial stenosis: clinical profile in a series of 134 patients in Spain]. AB - INTRODUCTION: Intracranial stenoses (IS) are known to be a manifestation of atherosclerosis and a cause of cerebral ischemia, although very few clinical reports have appeared describing such patients in our milieu. OBJECTIVE: The aim of this study was to describe the vascular risk factors, clinical presentation, radiological characteristics, aetiological role played in strokes and the vessels affected in a series of Spanish patients suffering from stroke and IS. PATIENTS AND METHODS: We conducted a retrospective descriptive study of patients admitted to the Cerebrovascular Pathology Unit with strokes and IS between 1990 and 2001. Data collected included: age, sex, arterial hypertension (AHT), diabetes (Db), hypercholesterolemia (HC), smoking (Sm), ischemic or emboligenic heart disease, intermittent claudication, carotid atheromatosis, clinical presentation of stroke and earlier lesions in computerised axial tomography (CAT) scans of the brain. RESULTS: 132 patients; 187 stenotic vessels: 65.7% males, mean age 68.3 years. AHT 65%, Db 39%, HC 40%, Sm 43%, ischemic heart disease 22%, emboligenic heart disease 17%, intermittent claudication 13%. CLINICAL PRESENTATION: TIA 16%, LACI 33%; PACI 25%, TACI 4%, POCI 19%. Significant carotid atheromatosis 26.5%. Symptomatic IS 50%: mean age 63.3 years, 64% females. Arteries affected: vertebral (VA) 28%; middle cerebral (MCA) 27%; carotid siphon 21%; basilar (BA) 10% (65% symptomatic); anterior cerebral (ACA) 5% and posterior cerebral (CPA) 4%. Normal cranial CAT scan 24%, lacunar infarcts 42%, territorial 32%; leukoaraiosis 17%. CONCLUSIONS: Patients with stroke and IS display different clinical profiles according to their sex (males: a higher number of vascular risk factors and clinical involvement of other territories; females are more symptomatic, AHT and HC); they usually present clinically as lacunar syndromes, with a scarce amount of significant atheromatous carotid involvement, except IS of the VA, and IS of the BA are the most symptomatic. PMID- 15340887 TI - [Drug-cholinesterase-inhibitors persistence patterns in treated patients with dementia of Alzheimer type: retrospective comparative analysis of donepezil, rivastigmine and galantamine]. AB - INTRODUCTION AND OBJECTIVE: Long term duration of treatment of the dementia of Alzheimer type, the most frequent type of dementia in our environment, is associated with delay in patient functional and cognitive impairment. The aim of the study was to determine retrospectively the persistence of treatment with donepezil, rivastigmine and galantamine in patients with dementia of the Alzheimer type in a population setting. PATIENTS AND METHODS: Retrospective study performed in Primary Care Health Centres. The study included patients who were treated between January, 2000 and September, 2003. Kaplan-Meier survival analysis was applied to assess pattern of treatment persistence. RESULTS: A total of 95 patients (78.8% female), with a mean age of 77.6 years (SD: 6.2) were included; 39 donepezil, 35 rivastigmine, and 21 galantamine, with a mean MMSE score of 10.3 +/- 7.8, 9.7 +/- 8.4 and 13.6 +/- 7.1, respectively (p = 0.256). Mean treatment duration was donepezil: 80.3 (SD: 7.7) weeks, rivastigmine: 52.4 (SD: 5.3) weeks and galantamine: 49.8 (SE: 4.0) weeks, p < or = 0.01. Median persistence of treatment was 129.3 weeks for donepezil, 59.1 for rivastigmine and 45.0 for galantamine, p < or = 0.001 in both cases. At week 52, 62% of donepezil-treated patients maintained the initial therapy, compared with 40% rivastigmine-treated and 33% galantamine-treated, p < 0.05. CONCLUSION: This retrospective study found that patients with dementia of the Alzheimer type treated with donepezil showed more persistence of treatment compared which those who received rivastigmine or galantamine. PMID- 15340888 TI - [Disorder profile in the Mini-Mental State Examination in patients with mild cognitive impairment]. AB - INTRODUCTION: Mild cognitive impairment (MCI) is a condition that can act as a risk factor for the development of dementia. AIMS: Our objective was to determine the profile of performance in the MMSE (Mini-Mental State Examination) in patients with MCI. PATIENTS AND METHODS: We studied the performance in the MMSE of 117 patients who satisfied the diagnostic criteria for MCI. The means and standard deviations (SD) were calculated and the sample was stratified according to the total score obtained in the MMSE; performance in each area of the MMSE was analysed for each group. RESULTS: Mean performance in the MMSE was 27.87, with a range of 23 to 30, and a SD of 1.75. The most compromised areas were memory and attention; there were mild alterations in temporal orientation. The ability to copy figures was seen to be affected in the groups with the best performances and this was maintained and increased slightly in the most affected levels. CONCLUSIONS: The mean value obtained is very similar to that of the most recent studies and is above the cut score for the MMSE for schooled populations (25/26). Our study reaffirms the idea that the MMSE is not sensitive to the early stages of impairment. The most strongly involved areas are memory and attention. We discuss the visual-praxis disorder that was found, and assigned to the structure of the MMSE and the bias inherent in the sample, with no confirmable incidence of schooling. PMID- 15340889 TI - [Performance of carriers and non-carriers of the E280A mutation for familial Alzheimer's disease in a naming test]. AB - INTRODUCTION: Early preclinical diagnosis is the greatest challenge faced by researchers into dementia. Cognitive, neuroanatomical, neurophysiological and genetic markers have been reported. One of the preclinical cognitive markers is anomia and it is often assessed using visual naming tests. AIMS: The aim of this study was to analyse the type of mistakes made in a visual naming test in a group of carriers and non-carriers of the E280A PS1 mutation. PATIENTS AND METHODS: The sample was made up of 91 participants who were genotyped for the E280A PS1 mutation and divided into three groups: non-carriers (n = 30), asymptomatic carriers (n = 39) and sick carriers (n = 22). Selection was performed using the Minimental and the Fast and EDG scales and mistakes in the CERAD naming test were classified. The types of mistakes taken into account were: no answer, visual, semantic, phonological, the whole for the part, and not related. RESULTS: There is a significant difference in the number of semantic errors between non-carriers and asymptomatic carriers; on comparing the three groups, no statistically significant differences were found in visual mistakes. CONCLUSIONS: Visual mistakes are a general characteristic, even in healthy subjects and, therefore, these errors did not provide any information that could be used to classify patients with or without dementia. Semantic mistakes can be considered as being a preclinical sign in familial Alzheimer's disease (FAD). Both visual and auditory naming tests must be applied when evaluating patients with FAD. PMID- 15340890 TI - [Stromal cell transplant in the 6-OHDA lesion model]. AB - INTRODUCTION: A good deal of evidence currently exists to show that transplanting foetal mesencephalic tissue can produce symptomatic benefits both in patients and in disease models. Nevertheless, the technical and ethical difficulties involved in obtaining enough suitable foetal cerebral tissue have been a serious obstacle to its application. Stromal cells derived from bone marrow, due to their potential capacity to generate different types of cells, could be an ideal source of material for cell restoration in neurodegenerative diseases. AIMS: Our aim was to evaluate the effect of transplanting stromal cells derived from bone marrow on the behaviour of 6-OHDA rats, when they are inserted into the striatum. MATERIAL AND METHODS: In this study we used rats with a lesion in the substantia nigra induced by 6-hydroxydopamine, divided into several experimental groups. Rotary activity induced by D-amphetamine (5 mg/kg, intraperitoneally) was evaluated before and throughout the three months following the transplant in all the experimental groups, except in the group of healthy controls. Hemiparkinsonian rats received a total of 350 000 foetal ventral mesencephalic cells and 8 x 10(4) stromal cells/microL, which were implanted in the striatum. RESULTS AND CONCLUSIONS: Animals with stromal cells transplanted in the body of the striatum significantly reduced the number of turns induced by amphetamine (p < 0.05); yet this reduction was not greater than that induced by foetal mesencephalic cell transplants. We were also unable to demonstrate any significant improvement in the motor skills of the forelimbs. PMID- 15340891 TI - [Ruptured aneurysms presenting with subdural hematoma: management without angiography]. AB - INTRODUCTION: Ruptured aneurysms on rare occasions cause subdural hematomas as described in literature. Sudden deterioration and coma is a common feature in those patients and a emergent surgical attitude is prompt required, even without confirmation with angiography. CASE REPORTS: We described three cases with acute subdural hematomas and little or no subarachnoid hemorrhage caused by ruptured aneurisms who presented with rapid neurologic deterioration. Urgent craniotomy and evacuation of the hematoma was performed without previous angiography in the three patients. In two patients the aneurysm was found during surgical exploration and subsequently clipped; in the remaining patient the aneurysm was embolized postoperatively. CONCLUSIONS: The occurrence of a subdural hematoma caused by the rupture of an intracranial aneurysm must be suspected in spontaneous subdural hematomas, especially in association with disproportioned conscious deterioration. All the three patients we report debuted with sudden conscious deterioration. If a ruptured aneurysm causing subdural hematoma is suspected, early surgical intervention is required even if angiography is not available. Severe neurological deficit and uncal herniation might still be reversible if provided decompression can be carried out in promptly. Angiography availability should not postpone surgery. Aneurysm presence should be ruled out whether by surgical exploration or by delayed angiography. Posterior communicating aneurysm are related to formation of subdural hematoma. PMID- 15340892 TI - [Compression of the internal carotid artery due to elongated styloid process]. AB - INTRODUCTION: Elongation of the styloid process has been reported as a source of pain in the head and neck region. However, transient ischemic attack (TIA) due to mechanical compression of the internal carotid artery (ICA) by an elongated styloid process has never been mentioned in the literature. CASE REPORT: Male aged 36 years, with a ten-year history of dull intermittent pain in the left side of the neck that worsened on turning the head, suffered a TIA in the silvian territory, ten days after a minor neck trauma. The eco-Doppler exploration demonstrated a compression of the ICA after rotation of the head. The arteriography revealed an stenosis of the cervical ICA by an external compression. The length of the elongated process in the orthopantomogram and the computed tomography was 35 mm. Angiotomography showed a closed relationship between the tip of the left elongated process and the stenosis of the ICA. Surgical styloidectomy was performed via an extraoral approach under general anaesthesia. The excised specimen measured 30 mm long. All symptoms were relieved with no recurrence during a three-year follow up period. CONCLUSION: TIA should be taken into account in the clinical feature associated to an elongated styloid process. Angiotomography is very reliable to define the anatomic relationship between the styloid process and the ICA. The treatment of choice is surgical shortening trough an external approach which provides an adequate control of the carotid artery. PMID- 15340893 TI - [L-2-hydroxyglutaric aciduria: clinical, biochemical and neuroradiological findings in two Venezuelan patients]. AB - INTRODUCTION: L-2-hydroxyglutaric aciduria is a rare inborn error of metabolism, autosomal recessive, identified in about 50 patients. The primary defect is still unknown. The clinical phenotype is variable. Affected individuals show slowly progressive neurodegenerative disorder with cerebellar ataxia and mental retardation. Pyramidal, and extrapyramidal signs, seizures and macrocephaly have been reported. All patients previously described show a pattern of subcortical leukoencephalopathy with nearly empty gyral cores and cerebellar atrophy in neuroimaging studies. The diagnosis is established by detection of increased levels of L-2-hydroxyglutaric acid in urine, plasma and cerebrospinal fluid. CASE REPORTS: We here describe two patients 7 and 9 years old, who presented psychomotor retardation, seizures, progressive cognitive deterioration, and pyramidal, extrapyramidal and cerebellar signs. Magnetic resonance scanning of the brain demonstrated a bilateral subcortical leukoencephalopathy pattern and areas of increased T2-weighted signal in the basal ganglia and cerebellar dentate nuclei. The analysis of organic acids in urine by gas chromatography/mass spectrometry showed elevated 2-hydroxyglutaric acid, 100% of it in the form of L enantiomer. CONCLUSION: The diagnostic consideration is based on clinical findings and typical neuroimaging pattern and is established by detection of L-2 hydroxyglutaric acid in body fluids. Subcortical white matter loss is an important clue to diagnosis. PMID- 15340894 TI - [Spontaneous intracranial hypotension: the value of isotope cisternoscintigraphy using 99mTc-DTPA]. AB - INTRODUCTION: Spontaneous intracranial hypotension (SIH) is an infrequent clinical entity that is found predominantly in young adults and is characterised by the presence of acute or subacute headaches that appear on standing up and subsides on lying down. CASE REPORT: A 35-year-old female with a three-month history of holocranial headaches, accompanied by dizziness and gait instability, which increase on standing up and diminish to a certain extent on lying down, associated to bilateral tinnitus. The patient's history included a slight strain made 4 months earlier, with a sudden non-irradiated pain in the back of the neck that got better spontaneously. On the basis of the clinical and radiological findings from an MRI scan of the head and neck, our service was asked to perform a cisternoscintigraphy. CONCLUSIONS: Isotope cisternoscintigraphy using 99mTc DTPA confirmed the diagnosis of the process and enabled us to locate the CSF leak. We therefore think it advisable to highlight its value in the diagnosis of SIH syndrome so as to be able to avoid the use of other invasive complementary explorations that entail a certain degree of morbidity and mortality. PMID- 15340895 TI - [Hair with a double crown and dilatation of Virchow-Robin spaces: a proposal for a new neurocutaneous association]. AB - INTRODUCTION: The primitive ectoderm is the common origin of the nervous system, the skin and related organs. These structures can therefore be affected simultaneously by aggressions that take place during embryogenesis involving the ectodermal layer. To date multiple neurocutaneous associations have been reported, some of which were catalogued as genetically determined syndromes or diseases. Thus, it becomes possible to recognise these entities at an early stage, to carry out a better therapeutic approach to the neurological pathology and even, in some cases, to establish a prognosis. CASE REPORTS: We present five male patients between 2 and 7 years of age in whom we found a common pattern of deviated psychomotor and behavioural development consisting of a hyperactivity and impulsivity disorder associated to retarded language acquisition. Common to all of them was the hair growing with a double crown and dilatation of the Virchow-Robin spaces seen in the neuroimaging studies. CONCLUSIONS: We propose a clinical-radiological association that, to our knowledge, has no equivalent in the literature, and we highlight the importance of knowing how to recognise both the neuropsychiatric symptoms and the skin features, as well as the characteristic neuroimaging findings of this group of patients. PMID- 15340896 TI - [Hypocretins and adenosine in the regulation of sleep]. AB - OBJECTIVES: To review the recent discovery of hypocretins (orexins) and their link to the pathophysiology of narcolepsy and the role of adenosine in the integration of brain metabolism and sleep. DEVELOPMENT: The importance of the functions carried out by the hypothalamus in the regulation of sleep and the waking state has been consolidated by the discovery of hypocretins and the role played by cerebral adenosine. Hypocretins are two peptides made up of 33 and 28 amino acids whose neurons are located predominantly in the lateral hypothalamus and surrounding regions. In the Doberman canine narcolepsy model, in which this disease is presented with an autosomal recessive pattern, a mutation was detected in one of the receptors involved in the hypocretin system, namely the hypocretin 2 receptor. Failures in the hypocretin system have been confirmed as a key factor in narcolepsy by other findings in laboratory animals and humans. Adenosine, on the other hand, is accumulated during the waking state as a result of neuronal metabolism and this in turn is related to drowsiness. Sleep episodes lower the levels of this substance in the brain. Adenosine receptor antagonists increase wakefulness (e.g. caffeine), while the agonists promote slow-wave sleep. CONCLUSIONS: Hypocretins and adenosine from the hypothalamus perform functions involving the regulation of sleep and wakefulness. Understanding these two systems can have repercussions on clinical problems such as insomnia, hypersomnia and other neuropsychiatric disorders. PMID- 15340897 TI - [The prognostic value of computerized tomography in acute aneurysmal subarachnoid haemorrhages]. AB - INTRODUCTION AND DEVELOPMENT: The radiological diagnosis of subarachnoid haemorrhages (SAH) is based fundamentally on studies using computerised tomography, although magnetic resonance imaging has also been proved to be highly sensitive. Spontaneous SAH is essentially due to the rupture of a cerebral aneurysm. The distribution of the SAH and the presence of associated haemorrhagic lesions are useful for developing a suspicion as to the exact whereabouts of the aneurysm causing the SAH. CONCLUSIONS: SAH is still linked to a poor prognosis despite the progress made in its diagnosis and treatment. Several radiological factors have been reported as being associated to a poorer prognosis: 1. The risk of complications is related to the extension of the SAH. The most widely accepted classification for determining the extension is the one put forward by Fisher. The higher grades III and IV are linked to a higher rate of morbidity and mortality. 2. SAH that is situated in the perimesencephalic cisterns is linked to a low risk of being of aneurysmal origin and to a good progression. 3. A size above 10 mm is linked to a poorer progression. 4. Finally, the appearance of any of the possible complications (vasospasm, rebleeding, haemorrhage, edema) is also linked to a poorer prognosis. PMID- 15340898 TI - [Painful neuropathies: their pathophysiology and treatment]. AB - OBJECTIVES: The purpose of this study is to review the different studies published in the literature concerning the different physiological mechanisms involved in the genesis of painful neuropathy, as well as the diagnostic options and the different pharmacological treatments currently available. DEVELOPMENT: Distinct pathologies usually condition painful neuropathy, one of the main ones being diabetes mellitus. The triggering phenomenon is often some kind of damage to the tissues that contain nervous pain receptors, which later gives rise to a release of proinflammatory molecules, and triggers a cascade of phenomena that result in disorders in the central and peripheral nervous system (peripheral and central sensitisation). These disorders usually produce clinical manifestations, such as allodynia, paresthesias, among others, and these are sometimes the sole manifestation of painful neuropathy. Diagnosis of this syndrome is at times complicated due to the involvement of thin fibres, which cannot be identified by the conventional methods used in neurophysiological studies. There is also a broad range of pharmaceuticals used in the treatment of painful neuropathy that range from tricyclic antidepressants, non-steroidal anti-inflammatory drugs, opioid analgesics, antiarrhythmics and even agents for topical use. CONCLUSIONS: Diagnosis of thin fibre neuropathy is usually performed by carrying out a Quantitative Sudomotor Axon Reflex Test, quantitative sensory tests and a skin biopsy. As regards the pharmacological treatment, the new generation of anticonvulsive drugs like gabapentin seems to have advantages over the traditional pharmaceuticals, although their widespread use is still largely restricted by their cost. PMID- 15340899 TI - [The safety and tolerability profile of gabapentin in optimal doses]. AB - INTRODUCTION: Gabapentin (GBP) is an amino acid that is structurally analogous to GABA with the capacity to cross the blood-brain barrier and an antiepileptic action. It is indicated in the treatment of partial seizures with or without secondary generalisation. It has been suggested that the dosages used in pilot studies were not suitable to achieve optimal efficiency. AIMS: The aims of this study were to investigate the safety and tolerability of GBP with doses above those initially advocated in the literature. DEVELOPMENT: We review its pharmacological characteristics and analyse the results of different placebo controlled, open, short and long-term trials, both in monotherapy and associated to other antiepileptic drugs. CONCLUSION: Enough evidence was found to allow us to state that the efficiency of GBP can be increased with the utilisation of higher doses than those recommended to date, without the risk of generating unacceptable side effects at the same time. PMID- 15340900 TI - [The pharmacology of vestibular disorders]. AB - INTRODUCTION: In recent years, significant advances have been made in the physiology and the pharmacology of vestibular disorders. It is now possible to elaborate an approximation of the mechanisms of action of the drugs used in the symptomatic treatment of vertigo. DEVELOPMENT: These drugs usually have a number of different pharmacological actions and overlapping effects. They can modify the intensity of the symptoms (e.g. vestibular suppressants) or prevent recurrences (e.g. calcium blockers in the case of vestibular migraine). Most of the drugs used for the acute treatment of vertigo can have a negative effect on the central compensation mechanisms. Vestibular rehabilitation exercises and certain drugs can speed up compensation. Choice of medication and the method of administration depend on the severity of the clinical symptoms, the pattern of temporal evolution, the underlying vestibular disease, the associated clinical conditions and the profile of the collateral effects of the drugs utilised. CONCLUSIONS: Vestibular suppressants and antiemetic drugs are still the basis of the acute treatment of vertigo. PMID- 15340901 TI - [The effectiveness and safety of botulinum toxin in the treatment of chronic primary headaches]. AB - INTRODUCTION: Primary headaches (PH), which are highly prevalent pathologies, are linked to high morbidity and disability rates and high costs for the health care system. Due to the partial effectiveness of prophylactic treatments, especially in chronic headaches, or the existence of contraindications that make them unsuitable for prescription, the use of botulinum toxin (BTX) arises as a promising preventive therapy. AIMS: The purpose of this study was to determine the usefulness and the mechanism of action of BTX in PH through a review of the literature. DEVELOPMENT: Recent results of BTX use in PH are contradictory and, although controlled studies with placebos are scarce, it has been suggested that it could have a grade II-III evidence level of effectiveness and safety, with greater benefits in refractory chronic migraine and chronic tension-type headache, together with the absence of analgesic abuse and psychiatric illness. The probable mechanism of action in PH is through a peripheral antinociceptive effect related to a decreased release of neuropeptides and to the transmission of impulses with indirect inhibition of nociceptive centres in the brain stem, which is an effect that is apparently intensified with repeated applications. A correlation has been claimed between the use of BTX and a reduction in the intensity, severity, number of headaches per month, diminished use of analgesics per month and lowered total management costs. Side effects of the treatment are infrequent (< 1%) and transient. CONCLUSIONS: Despite findings that have proved BTX to be effective and safe as prophylactic therapy in PH, it is still not considered to be the first choice preventive treatment. Further studies are needed to establish sites, doses and application schemas (fixed vs. mobile), as well as a subtype of patients who can benefit from its use. PMID- 15340902 TI - [Nervous system tuberculosis in state hospitals in Honduras]. PMID- 15340903 TI - [Thrombosis of the cerebral venous sinuses associated to ulcerative colitis. A case report]. PMID- 15340904 TI - [Hydranencephaly. Findings from computerised axial tomography and magnetic resonance scans]. PMID- 15340905 TI - [Bacterial meningitis following spinal anaesthesia]. PMID- 15340906 TI - [Acyclovir neurotoxicity vs. encephalitis due to varicella zoster in a patient with chronic renal failure]. PMID- 15340907 TI - Smart materials based on self-assembled hydrogen-bonded comb-shaped supramolecules. AB - Block copolymer self-assembly and supramolecular chemistry can be combined most naturally to prepare smart polymer nanomaterials. An attractive route is based on comb-shaped supramolecules, obtained by attaching side chains to (co)polymers by physical (non-covalent) interactions. Hydrogen bonding is a key element of our approach. It combines an ease of synthesis with other important approach-specific elements, such as hierarchical self-assembly, strongly enhanced processability, swelling, and cleaving. Functional properties discussed include anisotropic proton conductivity, switching proton conductivity, electronically conducting nanowires, polarized luminance, dielectric stacks (optical reflectivity), functional membranes, and nano objects. PMID- 15340908 TI - Development of catalytic asymmetric reactions via chiral palladium enolates. AB - This article describes the generation of chiral palladium enolates and their application to several kinds of catalytic asymmetric reactions. Two methods to generate chiral enolates were developed using novel cationic palladium complexes 1 and 2. In these processes, water or a hydroxo ligand on palladium metal plays an important role as a nucleophile to promote the transmetallation or as a Bronsted base to abstract an acidic alpha-proton of the carbonyl group. These enolates showed sufficient reactivity with various electrophiles. Using a chiral Pd enolate as a key intermediate, highly enantioselective reactions such as catalytic aldol reactions, Mannich-type reactions, Michael reactions, and fluorination reactions were developed. The unique structures of the palladium enolate complexes were elucidated and reaction mechanisms are proposed. PMID- 15340909 TI - Comparative chemistry of isolable divalent compounds of silicon, germanium, and tin. AB - Recent studies of the synthesis, structures, spectroscopic properties, and reactions of a series of isolable metallylenes (R2E:, E = Si (1), Ge (2), and Sn (3); R2 = 1,1,4,4-tetrakis(trimethylsilyl)butane-1,4-diyl) are summarized. Because these group-14 metallylenes bear the same helmet-like ligand, a straightforward discussion of the element-dependence of the intrinsic properties of the group-14 element divalent compounds is possible. All these metallylenes were monomeric both in solution and in the solid state, indicating the effective steric protection by the ligand against dimerization. A small sigma-pi conjugation between C-Si(substituent) sigma orbitals and the vacant npz orbitals of divalent atoms in R2E: exists and the extent decreases in the order E = Si > Ge > Sn, as evidenced by UV-vis and NMR spectroscopies and X-ray crystallography. However, the extent of the sigma-pi conjugation in metallylenes 1-3 was much smaller than the electron-donating effects of neighboring nitrogen atoms in known stable cyclic diamino-substituted metallylenes, and hence metallylenes 1-3 are regarded as the least electronically perturbed. Comparative studies of the unique reactions among these metallylenes are also discussed. PMID- 15340910 TI - Biochemical reaction engineering for redox reactions. AB - Redox reactions are still a challenge for biochemical engineers. A personal view for the development of this field is given. Cofactor regeneration was an obstacle for quite some time. The first technical breakthrough was achieved with the system formate/formate dehydrogenase for the regeneration of NADH2. In cases where the same enzyme could be used for chiral reduction as well as for cofactor regeneration, isopropanol as a hydrogen source proved to be beneficial. The coproduct (acetone) can be removed by pervaporation. Whole-cell reductions (often yeast reductions) can also be used. By proper biochemical reaction engineering, it is possible to apply these systems in a continuous way. By cloning a formate dehydrogenase and an oxidoreductase "designer bug" can be obtained where formate is used instead of glucose as the hydrogen source. Complex sequences of redox reactions can be established by pathway engineering with a focus on gene overexpression or with a focus on establishing non-natural pathways. The success of pathway engineering can be controlled by measuring cytosolic metabolite concentrations. The optimal exploitation of such systems calls for the integrated cooperation of classical and molecular biochemical engineering. PMID- 15340911 TI - Comparative evaluation of eight docking tools for docking and virtual screening accuracy. AB - Eight docking programs (DOCK, FLEXX, FRED, GLIDE, GOLD, SLIDE, SURFLEX, and QXP) that can be used for either single-ligand docking or database screening have been compared for their propensity to recover the X-ray pose of 100 small-molecular weight ligands, and for their capacity to discriminate known inhibitors of an enzyme (thymidine kinase) from randomly chosen "drug-like" molecules. Interestingly, both properties are found to be correlated, since the tools showing the best docking accuracy (GLIDE, GOLD, and SURFLEX) are also the most successful in ranking known inhibitors in a virtual screening experiment. Moreover, the current study pinpoints some physicochemical descriptors of either the ligand or its cognate protein-binding site that generally lead to docking/scoring inaccuracies. PMID- 15340912 TI - Modeling correlated main-chain motions in proteins for flexible molecular recognition. AB - We describe a new method for modeling protein and ligand main-chain flexibility, and show its ability to model flexible molecular recognition. The goal is to sample the full conformational space, including large-scale motions that typically cannot be reached in molecular dynamics simulations due to the computational intensity, as well as conformations that have not been observed yet by crystallography or NMR. A secondary goal is to assess the degree of flexibility consistent with protein-ligand recognition. Flexibility analysis of the target protein is performed using the graph-theoretic algorithm FIRST, which also identifies coupled networks of covalent and noncovalent bonds within the protein. The available conformations of the flexible regions are then explored with ROCK by random-walk sampling of the rotatable bonds. ROCK explores correlated motions by only sampling dihedral angles that preserve the coupled bond networks in the protein and generates conformers with good stereochemistry, without using a computationally expensive potential function. A representative set of the conformational ensemble generated this way can be used as targets for docking with SLIDE, which handles the flexibility of protein and ligand side chains. The realism of this protein main-chain conformational sampling is assessed by comparison with time-resolved NMR studies of cyclophilin A motions. ROCK is also effective for modeling the flexibility of large cyclic and polycyclic ligands, as demonstrated for cyclosporin and zearalenol. The use of this combined approach to perform docking with main-chain flexibility is illustrated for the cyclophilin A-cyclosporin complex and the estrogen receptor in complex with zearalenol, while addressing the question of how much flexibility is allowed without hindering molecular recognition. PMID- 15340913 TI - Toward an atomistic model for predicting transcription-factor binding sites. AB - Identifying the specific DNA-binding sites of transcription-factor proteins is essential to understanding the regulation of gene expression in the cell. Bioinformatics approaches are fast compared to experiments, but require prior knowledge of multiple binding sites for each protein. Here, we present an atomistic force-field method to predict binding sites based only on the X-ray structure of a related bound complex. Specific flexible contacts between the protein and DNA are modeled by a library of amino acid side-chain rotamers. Using the example of the mouse transcription factor, Zif268, a well-studied zinc-finger protein, we show that the protein sequence alone, without the detailed experimental structure, gives a strong bias toward the consensus binding site. PMID- 15340914 TI - Structure of an anti-DNA fab complexed with a non-DNA ligand provides insights into cross-reactivity and molecular mimicry. AB - Antibodies that recognize DNA (anti-DNA) are part of the autoimmune response underlying systemic lupus erythematosus. To better understand molecular recognition by anti-DNA antibodies, crystallographic studies have been performed using an anti-ssDNA antigen-binding fragment (Fab) known as DNA-1. The previously determined structure of a DNA-1/dT5 complex revealed that thymine bases insert into a narrow groove, and that ligand recognition primarily involves the bases of DNA. We now report the 1.75-A resolution structure of DNA-1 complexed with the biological buffer HEPES (4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid). All three light chain complementarity-determining regions (CDRs) and HCDR3 contribute to binding. The HEPES sulfonate hydrogen bonds to His L91, Asn L50, and to the backbone of Tyr H100 and Tyr H100A. The Tyr side-chains of L32, L92, H100, and H100A form nonpolar contacts with the HEPES ethylene and piperazine groups. Comparison to the DNA-1/dT5 structure reveals that the dual recognition of dT5 and HEPES requires a 13-A movement of HCDR3. This dramatic structural change converts the combining site from a narrow groove, appropriate for the edge-on insertion of thymine bases, to one sufficiently wide to accommodate the HEPES sulfonate and piperazine. Isothermal titration calorimetry verified the association of HEPES with DNA-1 under conditions similar those used for crystallization (2 M ammonium sulfate). Interestingly, the presence of 2 M ammonium sulfate increases the affinities of DNA-1 for both HEPES and dT5, suggesting that non-polar Fab-ligand interactions are important for molecular recognition in highly ionic solvent conditions. The structural and thermodynamic data suggest a molecular mimicry mechanism based on structural plasticity and hydrophobic interactions. PMID- 15340915 TI - Efficiency of a second-generation HIV-1 protease inhibitor studied by molecular dynamics and absolute binding free energy calculations. AB - A subnanomolar inhibitor of human immunodeficiency virus type 1 (HIV-1) protease, designated QF34, potently inhibits the wild-type and drug-resistant enzyme. To explain its broad activity, the binding of QF34 to the wild-type HIV-1 protease is investigated by molecular dynamics simulations and compared to the binding of two inhibitors that are used clinically, saquinavir (SQV) and indinavir (IDV). Analysis of the flexibility of protease residues and inhibitor segments in the complex reveals that segments of QF34 were more mobile during the dynamics studies than the segments of SQV and IDV. The dynamics of hydrogen bonding show that QF34 forms a larger number of stable hydrogen bonds than the two inhibitors that are used clinically. Absolute binding free energies were calculated with molecular mechanics-generalized Born surface area (MM-GBSA) methodology using three protocols. The most consistent results were obtained using the single trajectory approach, due to cancellation of errors and inadequate sampling in the separate-trajectory protocols. For all three inhibitors, energy components in favor of binding include van der Waals and electrostatic terms, whereas polar solvation and entropy terms oppose binding. Decomposition of binding energies reveals that more protease residues contribute significantly to the binding of QF34 than to the binding of SQV and IDV. Moreover, contributions from protease main chains and side chains are balanced in the case of QF34 (52:48 ratio, respectively), whereas side chain contributions prevail in both SQV and IDV (main chain:side-chain ratios of 41:59 and 45:55, respectively). The presented results help explain the ability of QF34 to inhibit multiple resistant mutants and should be considered in the design of broad-specificity second-generation HIV-1 protease inhibitors. PMID- 15340916 TI - Rational proteomics II: electrostatic nature of cofactor preference in the short chain oxidoreductase (SCOR) enzyme family. AB - The dominant role of long-range electrostatic interatomic interactions in nicotinamide adenine dinucleotide/nicotinamide adenine dinucleotide phosphate (NAD/NADP) cofactor recognition has been shown for enzymes of the short-chain oxidoreductase (SCOR) family. An estimation of cofactor preference based only on the contribution of the electrostatic energy term to the total energy of enzyme cofactor interaction has been tested for approximately 40 known three-dimensional (3D) crystal complexes and approximately 330 SCOR enzymes, with cofactor preference predicted by the presence of Asp or Arg recognition residues at specific 3D positions in the beta2alpha3 loop (Duax et al., Proteins 2003;53:931 943). The results obtained were found to be consistent with approximately 90% reliable cofactor assignments for those subsets. The procedure was then applied to approximately 170 SCOR enzymes with completely uncertain NAD/NADP dependence, due to the lack of Asp and Arg marker residues. The proposed 3D electrostatic approach for cofactor assignment ("3D_DeltaE(el)") has been implemented in an automatic screening procedure, and together with the use of marker residues proposed earlier (Duax et al., Proteins 2003;53:931-943), increases the level of reliable predictions for the putative SCORs from approximately 70% to approximately 90%. It is expected to be applicable for any NAD/NADP-dependent enzyme subset having at least 25-30% sequence identity, with at least one enzyme of known 3D crystal structure. PMID- 15340917 TI - Computational, spectroscopic, and resonant mirror biosensor analysis of the interaction of adrenodoxin with native and tryptophan-modified NADPH-adrenodoxin reductase. AB - In steroid hydroxylation system in adrenal cortex mitochondria, NADPH-adrenodoxin reductase (AR) and adrenodoxin (Adx) form a short electron-transport chain that transfers electrons from NADPH to cytochromes P-450 through FAD in AR and [2Fe 2S] cluster in Adx. The formation of [AR/Adx] complex is essential for the electron transfer mechanism in which previous studies suggested that AR tryptophan (Trp) residue(s) might be implicated. In this study, we modified AR Trps by N-bromosuccinimide (NBS) and studied AR binding to Adx by a resonant mirror biosensor. Chemical modification of tryptophans caused inhibition of electron transport. The modified protein (AR*) retained the native secondary structure but showed a lower affinity towards Adx with respect to AR. Activity measurements and fluorescence data indicated that one Trp residue of AR may be involved in the electron transferring activity of the protein. Computational analysis of AR and [AR/Adx] complex structures suggested that Trp193 and Trp420 are the residues with the highest probability to undergo NBS-modification. In particular, the modification of Trp420 hampers the correct reorientation of AR* molecule necessary to form the native [AR/Adx] complex that is catalytically essential for electron transfer from FAD in AR to [2Fe-2S] cluster in Adx. The data support an incorrect assembly of [AR*/Adx] complex as the cause of electron transport inhibition. PMID- 15340918 TI - Gibbs adsorption isotherm combined with Monte Carlo sampling to see action of cosolutes on protein folding. AB - Driven by conditions set by smaller solutes, proteins fold and unfold. Experimentally, these conditions are stated as intensive variables--pH and other chemical potentials--as though small solutes were infinite resources that come at an externally varied free energy cost. Computationally, the finite spaces of simulation allow only fixed numbers of these solutes. By combining the analytic Gibbs adsorption isotherm with the computational Monte Carlo sampling of polymer configurations, we have been able to overcome an inherent limitation of computer simulation. The idea is to compute analytically the free energy changes wrought by solutes on each particular configuration. Then numerical computation is needed only to sample the set of configurations as efficiently as when no bathing solute is present. For illustration, the procedure is applied to an idealized two dimensional heteropolymer to yield lessons about the effect of cosolutes on protein stability. PMID- 15340919 TI - Improved prediction for N-termini of alpha-helices using empirical information. AB - The prediction of the secondary structure of proteins from their amino acid sequences remains a key component of many approaches to the protein folding problem. The most abundant form of regular secondary structure in proteins is the alpha-helix, in which specific residue preferences exist at the N-terminal locations. Propensities derived from these observed amino acid frequencies in the Protein Data Bank (PDB) database correlate well with experimental free energies measured for residues at different N-terminal positions in alanine-based peptides. We report a novel method to exploit this data to improve protein secondary structure prediction through identification of the correct N-terminal sequences in alpha-helices, based on existing popular methods for secondary structure prediction. With this algorithm, the number of correctly predicted alpha-helix start positions was improved from 30% to 38%, while the overall prediction accuracy (Q3) remained the same, using cross-validated testing. Although the algorithm was developed and tested on multiple sequence alignment based secondary structure predictions, it was also able to improve the predictions of start locations by methods that use single sequences to make their predictions. Furthermore, the residue frequencies at N-terminal positions of the improved predictions better reflect those seen at the N-terminal positions of alpha-helices in proteins. This has implications for areas such as comparative modeling, where a more accurate prediction of the N-terminal regions of alpha helices should benefit attempts to model adjacent loop regions. The algorithm is available as a Web tool, located at http://rocky.bms.umist.ac.uk/elephant. PMID- 15340920 TI - Glycine N-methyltransferases: a comparison of the crystal structures and kinetic properties of recombinant human, mouse and rat enzymes. AB - Glycine N-methyltransferases (GNMTs) from three mammalian sources were compared with respect to their crystal structures and kinetic parameters. The crystal structure for the rat enzyme was published previously. Human and mouse GNMT were expressed in Escherichia coli in order to determine their crystal structures. Mouse GNMT was crystallized in two crystal forms, a monoclinic form and a tetragonal form. Comparison of the three structures reveals subtle differences, which may relate to the different kinetic properties of the enzymes. The flexible character of several loops surrounding the active site, along with an analysis of the active site boundaries, indicates that the observed conformations of human and mouse GNMTs are more open than that of the rat enzyme. There is an increase in kcat when going from rat to mouse to human, suggesting a correlation with the increased flexibility of some structural elements of the respective enzymes. PMID- 15340921 TI - Significance of conformational biases in Monte Carlo simulations of protein folding: lessons from Metropolis-Hastings approach. AB - Despite significant effort, the problem of predicting a protein's three dimensional fold from its amino-acid sequence remains unsolved. An important strategy involves treating folding as a statistical process, using the Markov chain formalism, implemented as a Metropolis Monte Carlo algorithm. A formal prerequisite of this approach is the condition of detailed balance, the plausible requirement that at equilibrium, the transition from state i to state j is traversed with the same probability as the reverse transition from state j to state i. Surprisingly, some relatively successful methods that use biased sampling fail to satisfy this requirement. Is this compromise merely a convenient heuristic that results in faster convergence? Or, is it instead a cryptic energy term that compensates for an incomplete potential function? I explore this question using Metropolis-Hasting Monte Carlo simulations. Results from these simulations suggest the latter answer is more likely. PMID- 15340922 TI - Kinetics of folding and unfolding of goat alpha-lactalbumin. AB - The equilibrium unfolding and the kinetic folding and unfolding of goat alpha lactalbumin (GLA) were studied by near- and far-ultraviolet circular dichroism (CD) and by stopped-flow fluorescence spectroscopy. Specifically, the influence of environmental conditions such as pH and Ca2+ binding was examined. Compared to the apo-form, the Ca2+-bound form was found to be strongly stabilized in equilibrium conditions at pH 7.5 and 25 degrees C. The kinetics of the refolding of apo-GLA show a major change of fluorescence intensity during the experimental dead-time, but this unresolved effect is strongly diminished in holo-GLA. In both cases, however, the chevron plots can adequately be fitted to a three-state model. Moreover, double-mix stopped-flow experiments showed that the native state (N) is reached through one major pathway without the occurrence of alternative tracks. In contrast to the homologous bovine alpha-lactalbumin (BLA), the compactness of GLA is strongly influenced by the presence of Ca2+ ions. Unlike the two-state transition observed in guanidine hydrochloride (GdnHCl)-induced equilibrium denaturation experiments at higher pH, an equilibrium intermediate state (I) is involved in denaturation at pH 4.5. In the latter case, analysis of the kinetic data makes clear that the intermediate and the unfolded states (U) show practically no Gibbs free energy difference and that they are in rapid equilibrium with each other. A possible explanation for these variations in stability and in folding characteristics with pH could be the degree of protonation of His107 that directly influences non-native interactions. Variation of environmental conditions and even small differences in sequence, therefore, can result in important effects on thermodynamic and folding parameters. PMID- 15340923 TI - Molecular modeling of the core of Abeta amyloid fibrils. AB - Amyloid fibrils, a key pathological feature of Alzheimer's disease (AD) and other amyloidosis implicated in neurodegeneration, have a characteristic cross-beta structure. Here we present a structural model for the core of amyloid fibrils formed by the Abeta peptide using computational approaches and experimental data. Abeta(15-36) was threaded against the parallel beta-helical proteins. Our multi layer model was constructed using the top scoring template 1lxa, a left-handed parallel beta-helical protein. This six-rung helical model has in-register repeats of the Abeta(15-36) sequence. Each rung has three beta-strands separated by two turns. The model was tested using molecular dynamics simulations in explicit water, and is in good agreement with a number of experimental observations. In addition, a model based on right-handed helical proteins is also described. The core structural model described here might serve as the building block of the Abeta(1-40) amyloid fibril as well as some other amyloid fibrils. PMID- 15340924 TI - Toward the detection and validation of repeats in protein structure. AB - We present a method called DAVROS to detect, localize, and validate repeating motifs in protein structure allowing for insertions and deletions. DAVROS uses the score matrix from a structural alignment program (SAP) to search for repeating motifs using an algorithm based on concepts from signal processing and the statistical properties of the alignments. The method was tested against a nonredundant Protein Data Bank, and each chain was assigned a score. For the top 50 chains ranked by score, 70% contain repeating motifs detected without error. These represent 14 types of fold covering alpha, beta, and alphabeta protein classes. A second data set comprising protein chains in different sequence families for triosephosphate isomerase (TIM) barrel, leucine-rich repeat (LRR), trefoil, and alpha-alpha barrel folds was used to assess the ability of DAVROS to detect all motifs within a specific fold. For the second test set, the percentage of motifs detected was highest for the LRR chains (88.7%) and least for the TIM barrels (60%). This variability results from the regularity of the LRR motif compared to the alphabeta units of the TIM barrel, which generally have many more indels. These reduce the strength of the repeat signal in the SAP matrix, making repeat detection more difficult. PMID- 15340925 TI - Detecting local structural similarity in proteins by maximizing number of equivalent residues. AB - A new algorithm for superimposing protein structures based on maximizing the number of spatially equivalent residues is introduced. The algorithm works in three distinct steps. First, the optimal residue map is calculated by structural alignment. By default, the double dynamic programming algorithm, as implemented in the program ASH, was used for the structure alignment step, but we also present results based on alignments imported from three other programs (Dali, CE, and VAST).Second, the structures are spatially superimposed such that the effective number of equivalent residues (NER)--aligned residue pairs that can be spatially overlapped--is maximized. The NER score is an analytic, differentiable similarity function that rewards spatially equivalent residues but ignores non equivalent ones. Maximization of the NER score results in accurate superpositions in cases where root mean square deviation (RMSD) minimization fails. Third, the NER function is used in conjunction with traditional dynamic programming to realign the structures based on the proximity of residues in the superposition. Results are presented for a wide range of superposition problems and compared to results from Dali, CE, and VAST. In addition, several structure-structure pairs that show only partial similarity are discussed, and results are compared to those from the LGA, SARF2, and ThreeCa programs. PMID- 15340926 TI - Patterns of retinal light absorption related to retinitis pigmentosa mutants from in silico model structures of rhodopsin. AB - Changes induced by mutations in rhodopsin that are associated with the degenerative visual disease retinitis pigmentosa result in an altered pattern of light absorption according to quantum mechanical simulations and reference experimental works. Eleven single-point mutations associated with retinitis pigmentosa at and in the proximity to the retinal binding pocket of rhodopsin have been modeled in silico and their spectra calculated with the NDOL (Neglect of Differential Overlap accounting L azimuthal quantum number) a priori method. The altered pattern of absorption found would lead to cumulative consequences in energy dissipation with aging. Different energy balances in the case of mutants at the very molecular level, compared to native nonmutated rhodopsin, can cause permanent cellular stress and would play a role in the progression of the retine degenerative process. It could explain the worsening of the pathological condition mostly in adults and suggests the probable beneficial effects of using quenching drugs and protection devices against excess of light in the early stages of life for avoiding or reducing potential damage. PMID- 15340927 TI - Large-scale prediction of protein geometry and stability changes for arbitrary single point mutations. AB - We have developed a method to both predict the geometry and the relative stability of point mutants that may be used for arbitrary mutations. The geometry optimization procedure was first tested on a new benchmark of 2141 ordered pairs of X-ray crystal structures of proteins that differ by a single point mutation, the largest data set to date. An empirical energy function, which includes terms representing the energy contributions of the folded and denatured proteins and uses the predicted mutant side chain conformation, was fit to a training set consisting of half of a diverse set of 1816 experimental stability values for single point mutations in 81 different proteins. The data included a substantial number of small to large residue mutations not considered by previous prediction studies. After removing 22 (approximately 2%) outliers, the stability calculation gave a standard deviation of 1.08 kcal/mol with a correlation coefficient of 0.82. The prediction method was then tested on the remaining half of the experimental data, giving a standard deviation of 1.10 kcal/mol and covariance of 0.66 for 97% of the test set. A regression fit of the energy function to a subset of 137 mutants, for which both native and mutant structures were available, gave a prediction error comparable to that for the complete training set with predicted side chain conformations. We found that about half of the variation is due to conformation-independent residue contributions. Finally, a fit to the experimental stability data using these residue parameters exclusively suggests guidelines for improving protein stability in the absence of detailed structure information. PMID- 15340928 TI - Ion transit pathways and gating in ClC chloride channels. AB - ClC chloride channels possess a homodimeric structure in which each monomer contains an independent chloride ion pathway. ClC channel gating is regulated by chloride ion concentration, pH and voltage. Based on structural and physiological evidence, it has been proposed that a glutamate residue on the extracellular end of the selectivity filter acts as a fast gate. We utilized a new search algorithm that incorporates electrostatic information to explore the ion transit pathways through wild-type and mutant bacterial ClC channels. Examination of the chloride ion permeation pathways supports the importance of the glutamate residue in gating. An external chloride binding site previously postulated in physiological experiments is located near a conserved basic residue adjacent to the gate. In addition, access pathways are found for proton migration to the gate, enabling pH control at hyperpolarized membrane potentials. A chloride ion in the selectivity filter is required for the pH-dependent gating mechanism. PMID- 15340929 TI - X-ray structure of a membrane-bound beta-glycosidase from the hyperthermophilic archaeon Pyrococcus horikoshii. AB - The beta-glycosidase of the hyperthermophilic Archaeon Pyrococcus horikoshii is a membrane-bound enzyme with the preferred substrate of alkyl-beta-glycosides. In this study, the unusual structural features that confer the extreme thermostability and substrate preferences of this enzyme were investigated by X ray crystallography and docking simulation. The enzyme was crystallized in the presence of a neutral surfactant, and the crystal structure was solved by the molecular replacement method and refined at 2.5 A. The main-chain fold of the enzyme belongs to the (betaalpha)8 barrel structure common to the Family 1 glycosyl hydrolases. The active site is located at the center of the C-termini of the barrel beta-strands. The deep pocket of the active site accepts one sugar unit, and a hydrophobic channel extending radially from there binds the nonsugar moiety of the substrate. The docking simulation for oligosaccharides and alkylglucosides indicated that alkylglucosides with a long aliphatic chain are easily accommodated in the hydrophobic channel. This sparingly soluble enzyme has a cluster of hydrophobic residues on its surface, situated at the distal end of the active site channel and surrounded by a large patch of positively charged residues. We propose that this hydrophobic region can be inserted into the membrane while the surrounding positively charged residues make favorable contacts with phosphate groups on the inner surface of the membrane. The enzyme could thus adhere to the membrane in the proximity of its glycolipid substrate. PMID- 15340930 TI - Nonlinear analysis of EEG during NREM sleep reveals changes in functional connectivity due to natural aging. AB - The spatial organization of nonlinear interactions between different brain regions during the first NREM sleep stage is investigated. This is achieved via consideration of four bipolar electrode derivations, Fp1F3, Fp2F4, O1P3, O2P4, which are used to compare anterior and posterior interhemispheric interactions and left and right intrahemispheric interactions. Nonlinear interdependence is detected via application of a previously written algorithm, along with appropriately generated surrogate data sets. It is now well understood that the output of neural systems does not scale linearly with inputs received and, thus, the study of nonlinear interactions in EEG is crucial. This approach also offers significant advantages over standard linear techniques, in that the strength, direction, and topography of the interdependencies can all be calculated and considered. Previous research has linked delta activity during the first NREM sleep stage to performance on frontally activating tasks during waking hours. We demonstrate that nonlinear mechanisms are the driving force behind this delta activity. Furthermore, evidence is presented to suggest that the aging brain calls upon the right parietal region to assist the pre-frontal cortex. This is highlighted by statistically significant differences in the rates of interdependencies between the left pre-frontal cortex and the right parietal region when comparing younger subjects (<23 years) with older subjects (>60 years). This assistance has been observed in brain-imaging studies of sleep deprived young adults, suggesting that similar mechanisms may play a role in the event of healthy aging. Additionally, the contribution to the delta rhythm via nonlinear mechanisms is observed to be greater in older subjects. PMID- 15340931 TI - Question/statement judgments: an fMRI study of intonation processing. AB - We examined changes in fMRI BOLD signal associated with question/statement judgments in an event-related paradigm to investigate the neural basis of processing one aspect of intonation. Subjects made judgments about digitized recordings of three types of utterances: questions with rising intonation (RQ; e.g., "She was talking to her father?"), statements with a falling intonation (FS; e.g., "She was talking to her father."), and questions with a falling intonation and a word order change (FQ; e.g., "Was she talking to her father?"). Functional echo planar imaging (EPI) scans were collected from 11 normal subjects. There was increased BOLD activity in bilateral inferior frontal and temporal regions for RQ over either FQ or FS stimuli. The study provides data relevant to the location of regions responsive to intonationally marked illocutionary differences between questions and statements. PMID- 15340932 TI - Brain-behavior correlation in children depends on the neurocognitive network. AB - We examined brain-behavior correlations in 12 children (age range 9.3 to 11.7 years) during a selective attention task that required the visual search of a conjunction of features and during a response inhibition task that required the inhibition of a pre-potent response during "no-go" blocks. We found that the association between performance in these tasks and brain activation as measured by functional magnetic resonance imaging (fMRI) depended on the neurocognitive network. Specifically, better performance during the no-go task was associated with greater activation in the response inhibition network including the prefrontal cortex and basal ganglia. In contrast, better performance during the visual search task was associated with less activation in the selective attention network including superior parietal lobule and lateral premotor cortex. These results show that the relation of performance to the magnitude of neural activation is complex and may display differential relationships based on the cognitive domain, anatomical region, and perhaps also developmental stage. PMID- 15340933 TI - Methods for detecting functional classifications in neuroimaging data. AB - Data-driven statistical methods are useful for examining the spatial organization of human brain function. Cluster analysis is one approach that aims to identify spatial classifications of temporal brain activity profiles. Numerous clustering algorithms are available, and no one method is optimal for all areas of application because an algorithm's performance depends on specific characteristics of the data. K-means and fuzzy clustering are popular for neuroimaging analyses, and select hierarchical procedures also appear in the literature. It is unclear which clustering methods perform best for neuroimaging data. We conduct a simulation study, based on PET neuroimaging data, to evaluate the performances of several clustering algorithms, including a new procedure that builds on the kth nearest neighbor method. We also examine three stopping rules that assist in determining the optimal number of clusters. Five hierarchical clustering algorithms perform best in our study, some of which are new to neuroimaging analyses, with Ward's and the beta-flexible methods exhibiting the strongest performances. Furthermore, Ward's and the beta-flexible methods yield the best performances for noisy data, and the popular K-means and fuzzy clustering procedures also perform reasonably well. The stopping rules also exhibit good performances for the top five clustering algorithms, and the pseudo T2 and pseudo-F stopping rules are superior for noisy data. Based on our simulations for both noisy and unscaled PET neuroimaging data, we recommend the combined use of the pseudo-F or pseudo-T2 stopping rule along with either Ward's or the beta-flexible clustering algorithm. PMID- 15340934 TI - Realistic spatial sampling for MEG beamformer images. AB - The spatial resolution achievable using magnetoencephalography (MEG) beamformer techniques is inhomogeneous across the brain and is related directly to the amplitude of the underlying electrical sources [Barnes and Hillebrand, Hum Brain Mapp 2003;18:1-12; Gross et al., Proc Natl Acad Sci USA 2001;98:694-699; Van Veen et al., IEEE Trans Biomed Eng 1997;44:867-860; Vrba and Robinson, Proc 12th Int Conf Biomagn 2001]. We set out to examine what an adequate level of spatial sampling of the brain volume is in a realistic situation, and what implications these inhomogeneities have for region-of-interest analysis. As a basis for these calculations, we used a simple retinotopic mapping experiment where stimuli were 17-Hz reversing gratings presented in either left or right visual hemifield. Beamformer weights were calculated based on the covariance of the MEG data in a 0 80 Hz bandwidth. We then estimated volumetric full-width half-maximum (FWHM) maps at a range of sampling levels. We show that approximately 10% of the 1 mm cubic voxels in the occipital volume have a FWHM smoothness of <5 mm, and 80% <10 mm in three subjects. This was despite relatively low mean signal-to-noise ratios (SNR) values of 1.5. We demonstrate how visualization of these FWHM maps can be used to avoid some of the pitfalls implicit in beamformer region-of-interest analysis. PMID- 15340943 TI - Theoretical chemistry of gold. AB - Gold is an element whose unique properties are strongly influenced by relativistic effects. A large body of appropriate calculations now exist and their main conclusions are summarized. The theoretical interpretation of the aurophilic attraction is discussed in detail. PMID- 15340944 TI - Efficient photocatalytic decomposition of organic contaminants over CaBi2O4 under visible-light irradiation. PMID- 15340945 TI - Artificial cells: temperature-dependent, reversible Li+-ion uptake/release equilibrium at metal oxide nanocontainer pores. PMID- 15340946 TI - Synthesis of diode molecules and their sequential assembly to control electron transport. PMID- 15340947 TI - Direct organocatalytic enantioselective mannich reactions of ketimines: an approach to optically active quaternary alpha-amino acid derivatives. PMID- 15340948 TI - A C3-symmetrical chiral trisoxazoline zinc complex as a functional model for zinc hydrolases: kinetic resolution of racemic chiral esters by transesterification. PMID- 15340949 TI - Stereoselective synthesis of a topologically chiral molecule: the trefoil knot. PMID- 15340950 TI - Asymmetric synthesis utilizing circularly polarized light mediated by the photoequilibrium of chiral olefins in conjunction with asymmetric autocatalysis. PMID- 15340951 TI - Mechanistic studies of a reaction promoted by the [YLi3[tris(binaphthoxide)]] complex: are three 1,1'-bi-2-naphthol units in a rare-earth-alkali-metal heterobimetallic complex necessary? PMID- 15340952 TI - Synthesis and structure of a dimetallated buckybowl: coordination of one [Cp*Ru]+ unit to each side of corannulene. PMID- 15340953 TI - A 1,3-diaza-2,4-distannacyclobutanediide: synthesis, structure, and bonding. PMID- 15340954 TI - Biomimetic oxidation of chromium(III): does the antidiabetic activity of chromium(III) involve carcinogenic chromium(VI)? PMID- 15340955 TI - Rhodizonate metal complexes with a 2D chairlike M6 metal-organic framework: [M(C6O6)(bpym)(H2O)].n H2O. PMID- 15340956 TI - The singlet excited state of a stable dialkylsilylene is responsible for its photoreactions. PMID- 15340957 TI - The isolation and crystal structure of a cyclic selenenate ester derived from Bis(2,6-diformyl-4-tert-butylphenyl)diselenide and its glutathione peroxidase like activity. PMID- 15340958 TI - Allosteric and electrostatic reprogramming of a ditopic ligand. PMID- 15340960 TI - Total synthesis of xerulinic acid. PMID- 15340959 TI - Catalytic growth of Au nanoparticles by NAD(P)H cofactors: optical sensors for NAD(P)+-dependent biocatalyzed transformations. PMID- 15340961 TI - A flexible, palladium-catalyzed indole and azaindole synthesis by direct annulation of chloroanilines and chloroaminopyridines with ketones. PMID- 15340962 TI - Efficient whole-cell biotransformation in a biphasic ionic liquid/water system. PMID- 15340964 TI - Variation in sulfation pattern of galactosaminoglycan containing proteoglycans is associated with the development of uterine leiomyoma. AB - The variations in sulfation patterns of glycosaminoglycans (GAGs) present in human normal myometrium and uterine leiomyoma were characterized following specific enzymic treatments and analysis by high-performance capillary electrophoresis (HPCE). Differences in the disaccharide composition of galactosaminoglycans (GalAGs) between normal myometrium and uterine leiomyoma were found. The results of this study suggest that the development of uterine leiomyoma is closely related not only to quantitative changes in the amount of GAGs but also in the sulfation of GalAGs. These differences may be attributed to increased synthesis of fibrotic connective tissue and deposition of collagen. PMID- 15340965 TI - Development and validation of a high-sensitivity liquid chromatography/tandem mass spectrometry (LC/MS/MS) method with chemical derivatization for the determination of ethinyl estradiol in human plasma. AB - An ultra-sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of oral contraceptive ethinyl estradiol (EE) was developed and validated over the curve range of 2.5-500 pg/mL using 1 mL of human plasma sample. Ethinyl estradiol and the internal standard, ethinyl estradiol tetra-deuterated (EE-d4), were extracted from the plasma matrix with methyl t butyl ether, derivatized with dansyl chloride and then back-extracted into hexane. The hexane phase was evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatographic separation was achieved on a Luna C(18) column (50 x 2 mm, 5 micro m) with an isocratic mobile phase of 20:80 (v/v) water:acetonitrile with 1% formic acid. The offline derivatization procedure introduced the easily ionizable tertiary amine function group to EE. This greatly improved analyte sensitivity in electrospray ionization and enabled us to achieve the desired lower limit of quantitation at 2.5 pg/mL. This high sensitivity method can be used for therapeutic drug monitoring or supporting bio-equivalence and drug-drug interaction studies in human subjects. PMID- 15340966 TI - Subminute and sensitive determination of the neurotransmitter serotonin in urine by capillary electrophoresis with laser-induced fluorescence detection. AB - In this work, a sub-minute and sensitive capillary electrophoresis with laser induced fluorescence (CE-LIF) method was developed for the analysis and quantitation of the neurotransmitter 5-hydroxytryptamine (5-HT) or serotonin in urine. The method involves precolumn derivatization with fluorescein isothiocyanate isomer I (FITC) using an excitation light from an argon ion laser of 488 nm and a 520 nm band pass emission filter. Different variables that affect derivatization (pH, FITC concentration, reaction time and temperature) and separation (buffer concentration, pH, applied voltage and injection time) were studied. The linear dynamic range obtained was between 0 and 188 nM with a detection limit of 16 nm with a RSD between 2 and 9%. The applicability of the proposed method was demonstrated by analysis of 5-HT in human urine, establishing a concentration of 57 nM in control urine. The method was validated by standard addition methodology. PMID- 15340967 TI - Efficiency of antidepressant drugs as monoamine reuptake inhibitors: analysis of the hydrophobicity influence using biopartitioning micellar chromatographic data. AB - The reuptake blockade of biogenic amines by antidepressants is related not only to their therapeutics effects, but also to their side effects and potential drug drug interactions. As an alternative to classical quantitative structure-activity relationships studies, in this work we propose different quantitative retention activity relationships (QRAR) models that are able to describe the monoamine reuptake inhibition by antidepressants. The retention of compounds is measured using a biopartitioning micellar chromatography (BMC) system that can simulate the same hydrophobic, electronic and steric molecular interactions as those that condition drug activity. Since all the compounds considered in this work are structurally related because all of them share the same molecular features as the corresponding basic pharmacophore, the results obtained show that there is a retention range in which antidepressants present the highest monoamine reuptake inhibitor potency. PMID- 15340968 TI - HPLC study of tyrosinase inhibition by thiopronine. AB - Thiopronine (N-2-mercaptopropionyl-glycine, NMPG) inhibits the o-dihydroxy phenolase activities of mushroom tyrosinase. When d,l-3-4-dihydroxyphenylalanine (DOPA) is employed as substrate, the inhibition was found to be a competitive type with K(i) of 0.95 micro m. We found in addition that thiopronine interacts with the enzymatic generated product (o-quinone) to form a colourless conjugate compound causing an apparent inhibition. These data suggest that thiopronine inhibits mushroom tyrosinase activity in two ways: (1) by forming an adduct with dopaquinone; and (2) by direct interaction with the enzyme probably towards the copper (II) present in the active site or cysteine-rich domains. This finding was indicated by the presence of a lag period prior to the attainment of an inhibited steady-state rate. Both lag period and steady-state rate were dependent on thiopronine and substrate concentrations. An increase of thiopronine concentration causes longer lag periods as well as a concomitant decrease in the tyrosinase activity. The presence of an excess of copper (II) reverses the inhibition exerted by thiopronine. PMID- 15340969 TI - Investigation on liquid chromatographic separation of basic compounds using silica column with aqueous/organic mobile phase containing triethylamine and acetic acid. AB - A high-performance liquid chromatography (HPLC) method using silica column eluted with aqueous solvent mobile phase containing triethylamine (TEA) and acetic acid (ACH) at trace percentages was characterized for the analysis of basic compounds. The key mechanism of this system is ion-exchange accompanying interaction of silanol groups. The increase in the ACH concentration in the mobile phase minimizes the ionization of the silanol group, leading to reduced retention time. However, the greater extent of ionization of silanol caused by the increase of TEA concentration helps to retain basic compounds in the column. Further, the protonated TEA that is positively charged also competes for the ionized silanol group with basic compounds, resulting in the modification of retention time. On the other hand, the retention becomes longer with increasing proportion of either organic or aqueous solvent in mobile phase, and partial replacement of methanol with acetonitrile. PMID- 15340970 TI - Identification of photoproducts from imazosulfuron by HPLC. AB - Photolysis of imazosulfuron was studied in aqueous solution under UV light. The reaction followed a pseudo-first-order kinetic with significant correlation coefficient. The major photodegradation products of imazosulfuron after irradiation under UV light were separated and tentatively identified by HPLC-MS analysis as (4,6-dimethoxypyrimidine-2-yl)aminocarbonylsulfamic acid, 4,6 dimethoxy-2-ureidopyrimidine and 2,2'-dichloro-[3,3'] bi [imidazo[1,2-a] pyridinyl]. The results indicate that different reaction pathways are followed: (1) cleavage of the sulfonylurea bridge; (2) desulfonylation, which can proceed either by a carbon-sulfur cleavage or a nitrogen-sulfur cleavage. A mechanism for the formation of the photoproducts is proposed. PMID- 15340971 TI - Variations in content and structure of glycosaminoglycans of the vitreous gel from different mammalian species. AB - The vitreous of all species is composed of essentially the same type of extracellular matrix macromolecules organized to a transparent gel. In this study, the composition and fi ne chemical structure of the glycosaminoglycans (GAGs) in the vitreous gel from sheep and goat were determined and compared with those of human and pig vitreous gels. The results showed that, in all examined species; hyaluronan (HA) was the predominant GAG, whereas chondroitin sulphate (CS) was the minor one. In the vitreous gel of the most relative species, i.e. sheep and goat, higher amounts of both of HA and CS were estimated as compared with pig and human tissues. The distribution of hydrodynamic sizes of HA and CS was significantly differed among different species. All HA preparations consisted of molecules with great variability in hydrodynamic sizes. The relative proportions of the large HA molecules (size >1.8 x 10(6) kDa) were significantly higher in sheep and goat as compared with human and pig vitreous gel. The length of CS chains was also of larger size in sheep and goat (50 and 58 kDa, respectively) than the respective chains in human and pig vitreous gel (38 and 28 kDa, respectively). The sulphation patterns of CS preparations were determined following enzymic treatments, HPLC and capillary electrophoretic analyses. The human vitreous-derived CS chains showed quite different sulphation profile than that of CS isolated from other species, since 4-sulphated disaccharides were identified as the dominant moiety. In conclusion, significant compositional and structural variations between the vitreous matrixes of different species at the GAG level were identified. The functional significance of these species-dependent variations is discussed. PMID- 15340972 TI - Quantitation of normal and formaldehyde-modified deoxynucleosides by high performance liquid chromatography/UV detection. AB - A sensitive and selective method was developed for the first time to quantify simultaneously the normal and formaldehyde (FA)-modified bases in human placental DNA treated with 100 ppm FA for 20 h at 37 degrees Celsius. Digestion of DNA to deoxynucleosides with DNase I, phosphodiesterase and alkaline phosphatase occurred in that order with centrifugation steps. The normal and FA-modified deoxynucleosides were then resolved from one another and reagent blank interferences to produce selective separation through high performance liquid chromatography-ultraviolet detection at 254 nm. A C(18) reversed-phase column facilitated the resolution using 5 mm ammonium acetate and a gradient of 0-6% methanol at fl ow rates of 0.3-1.4 mL/min before column cleaning. The lower quantifiable limits for deoxyadenosine, deoxyguanosine, deoxycytidine, thymidine, N(6)-hydroxymethyldeoxyadenosine (N(6)-dA), N(2)-hydroxymethyldeoxyguanosine (N(2)-dG) and N(4)-hydroxymethyldeoxycytidine (N(4)-dC) were 11, 7.6, 12, 15, 10, 10 and 22 pmol, respectively. The abundance order of the modified deoxynucleosides was N(6)-dA > N(2)-dG > N(4)-dC. dT did not form hydroxymethyl derivatives. The respective concentrations were about 6.0, 10.0 and 23 pmol of modified deoxynucleosides in 80 micro g of human placental DNA after treatment with 100 micro g/mL of formalin for 20 h at 37 degrees Celsius. The stabilities of N(6)-dA and N(2)-dG were much better at -20 degrees Celsius than at 25 degrees Celsius, where the respective halftimes were about 50.1 and 21.0 h. PMID- 15340973 TI - An HPLC assay of hydroxyl radicals by the hydroxylation reaction of terephthalic acid. AB - An HPLC assay for hydroxyl radicals is described. The hydroxyl radical was trapped by terephthalic acid (non-fluorescent), and 2-hydroxyl terephthalic acid (fluorescent) was quantitated by HPLC-fluorescence detection. At a terephthalic acid concentration of 4.25 mmol/L, the hydroxyl radical formed in the Fenton reaction was successfully assayed in the concentration range of hydrogen peroxide of 2.5-50 micro mol/L, where the concentration of Fe(II) was 50 micro mol/L. The fluorescence of 2-hydroxy terephthalate was stable at 24 h, and its detection limit by this method was 5 nmol/L (100 fmol). PMID- 15340974 TI - Influence of object spatial location and task complexity on children's use of their preferred hand depending on their handedness consistency. AB - The goal of the present study was to compare the use of the preferred versus the nonpreferred hand in children performing two tasks of different levels of complexity, in various positions in space. Right-handed and left-handed consistent children were compared to their inconsistent counterparts. The results showed that the general tendency to use one's preferred hand to grasp an object is more or less pronounced depending on the object's spatial location and the task's complexity. The children used their preferred hand more often even when the object was presented contralaterally, and did so to a greater extent during the more complex task. Inconsistent-handers were more likely to shift to using their nonpreferred hand when the object was presented to this hand's side. Finally, we found that the influence of handedness consistency on hand use for the spatial positions varied with the complexity of the task. These results favor a view of handedness as a dynamic process in which motor preferences interact with task demand, probably through task-related attention. PMID- 15340975 TI - Circadian- and temperature-specific effects of early deprivation on rat maternal care and pup development: short-term markers for long-term effects? AB - We compare the effects on pup body weight and on maternal care of 4-hr separation from dam and littermates on postnatal Days 1 to 14 (early deprivation, ED) under different thermal and circadian conditions. ED was performed at either 21 degrees C (Cold), or 32 degrees C (Warm), and either during the light or dark phase. The comparison group was nonhandling (NH), either under a nonreversed (Light) or reversed (Dark) cycle. At weaning, Cold ED pups were of lower body weight than Warm ED pups, and Warm ED pups were of lower body weight than NH pups. Light and Dark ED pups received high care at reunion relative to NH, and Cold ED pups received higher care at several hours postreunion relative to Warm ED and NH pups. We propose that reduced pup weight and increased maternal care are short term markers for the severity of Cold ED, and that this manipulation could therefore impact negatively on emotionality and cognition in adulthood. PMID- 15340976 TI - Effects of diet and handling on initiation of independent ingestion in rats. AB - The present study examined the effects of dietary manipulation on the age of onset of weaning in rat pups. In Experiment 1, female rats were placed on a standard chow (SC) or high-fat (HF) diet 1 week following mating. Pups were weighed daily from birth to Day 12, then animals were placed into specialized cages for separate recording of food intake of pups and dams. Pups were offered the same diet as their dam, and food intake and body weight were determined twice daily until Day 25. The results demonstrated that pups reared by dams fed the HF diet initiated independent ingestion on Day 16, approximately 24 hr before pups reared by dams fed the SC diet. There were no differences in body weight in pups across the two diets. While few differences were noted across diets in pups' or dams' behavior, HF pups appeared to demonstrate a delay in the establishment of circadian patterns of food intake. In Experiment 2, all dams were maintained on an SC diet until the day after parturition. At that time, dams and litters were placed into specialized cages and divided into four groups: HF/HF, HF/SC, SC/SC, and SC/HF (dam's diet/pup's diet, respectively). The results demonstrated that dams given the HF diet had pups that initiated food intake approximately 2 days before the pups of dams given the SC diet. In addition, pups offered the HF diet, independent of the dam's diet, initiated food intake approximately 0.8 days prior to pups offered the SC diet. Further, by Day 12, HF dams had pups that were heavier than SC dams. The results suggest that the onset of weaning in rats is affected by maternal diet and the weaning diet available to the pup. PMID- 15340977 TI - Development of play fighting in kindling-prone (FAST) and kindling-resistant (SLOW) rats: how does the retention of phenotypic juvenility affect the complexity of play? AB - Rats selectively bred for susceptibility to amygdala kindling (FAST) have been shown to retain neural and behavioral features of the juvenile phase into adulthood. In contrast, rats selectively bred for resistance to amygdala kindling (SLOW) are neurobehaviorally more typically adult. The development of play fighting in male and female rats of both selected lines was studied. Given the apparent association of juvenility and play often noted in the literature for mammals in general, it was predicted that the FAST rats should be more playful and be more likely to retain the juvenile tactics of play that lead to more prolonged and complex patterns of social contact. As expected, FAST rats initiated more playful attacks and were more likely to defend against attacks than SLOW rats as both juveniles and adults. Unexpectedly, however, both selected lines exhibited patterns of defense that reduced the likelihood of complex and prolonged social contact. Importantly, the two selected lines did so by very different means. The FAST rats did so by avoiding contact whereas the SLOW rats did so by responding in an adult-typical manner that blocks contact. That is, the FAST rats exaggerated the changes typically occurring at puberty whereas the SLOW rats, at all ages, responded in a more adult manner. These data suggest that the different components of play fighting do not change uniformly with changes in the neurobehavioral underpinnings of juvenility. PMID- 15340978 TI - Styrene-7,8-oxide. PMID- 15340979 TI - Sulfallate. PMID- 15340980 TI - Tamoxifen. PMID- 15340981 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD); "dioxin". PMID- 15340982 TI - Tetrachloroethylene (perchloroethylene). PMID- 15340983 TI - Tetrafluoroethylene. PMID- 15340984 TI - Tetranitromethane. PMID- 15340985 TI - Thioacetamide. PMID- 15340986 TI - Thiotepa. PMID- 15340987 TI - Thiourea. PMID- 15340988 TI - Thorium dioxide. PMID- 15340989 TI - Environmental tobacco smoke. PMID- 15340990 TI - Smokeless tobacco. PMID- 15340991 TI - Tobacco smoking. PMID- 15340992 TI - Toluene diisocyanate. PMID- 15340993 TI - o-Toluidine and o-toluidine hydrochloride. PMID- 15340994 TI - Toxaphene. PMID- 15340995 TI - Trichloroethylene. PMID- 15340996 TI - 2,4,6-Trichlorophenol. PMID- 15340997 TI - 1,2,3-Trichloropropane. PMID- 15340998 TI - Ultraviolet radiation related exposures: broad-spectrum ultraviolet (UV) radiation, UVA, UVB, UVC, solar radiation, and exposure to sunlamps and sunbeds. PMID- 15340999 TI - Urethane. PMID- 15341000 TI - Vinyl bromide. PMID- 15341001 TI - Vinyl chloride. PMID- 15341002 TI - 4-Vinyl-1-cyclohexene diepoxide. PMID- 15341003 TI - Vinyl fluoride. PMID- 15341004 TI - Wood dust. PMID- 15341005 TI - [An important point in training students]. PMID- 15341006 TI - [Work in an outpatient clinic as an important element in the preparation of otorhinolaryngologists for residency]. PMID- 15341007 TI - [How to improve learning and research activities at the chair of otorhinolaryngology]. PMID- 15341008 TI - [Alternative methods of anesthesia in adenotomy in children]. PMID- 15341009 TI - [Efficacy of physiomer spray after endonasal surgical interventions]. PMID- 15341010 TI - [Use of mucolytic ACC in the treatment of some ENT diseases]. PMID- 15341011 TI - [Cholesteatoma of the tympanic activity in a female patient with otosclerosis]. PMID- 15341012 TI - [Osteoma of the left frontal sinus and ethmoid bone invading the orbit and anterior cranial fossa]. PMID- 15341013 TI - [Acute stenosis of the upper trachea as a result of collar and mediastinal emphysema developing in the presence of asthmatic status]. PMID- 15341014 TI - [Laryngeal papillomatosis. Role of human papilloma virus, perspectives of the diagnosis and treatment]. PMID- 15341015 TI - [Cauterization in the area of the head and neck]. PMID- 15341016 TI - [Emerging or re-emerging infections--tuberculosis of the cervical lymph nodes]. PMID- 15341017 TI - Popular health education and venereal diseases in Croatia between two World Wars. AB - The article presents the research of popular health education on venereal diseases in Croatia between the World Wars. In the inter-war period, the traditional plain approach to popular health education was replaced with new, complex meth-ods, which became the basis for the modem work in this field. New social medicine ideas and new health politics, as well as the new founded institutions such as the School of Public Health in Zagreb and different anti venereal outpatient facilities, were crucial for changing popular health education after World War I. Based mostly on archival documents, this article explores popular health education as a vehicle for identification of attitudes and concepts within the medical community. Ambivalence in the perception of essential approaches towards popular heath education is elaborated on the ground of controversies within prominent medical representatives. With the support of new technologies, public health methods in the inter-war period matured in form and complexity. Despite various new methods, which made their way into different parts of everyday life, the subject matter of venereal diseases was treated through a limited number of methods due to the conservative attitudes of society, as well as resistance of many physicians. PMID- 15341018 TI - Edward B. Lewis. PMID- 15341019 TI - International pediatric cardiac assistance in Croatia: results of the 10 year program. PMID- 15341020 TI - Incidence of acute angle-closure glaucoma in Dalmatia, Southern Croatia. PMID- 15341021 TI - Occupational exposure and central nervous system tumors. PMID- 15341022 TI - Telomere length heterogeneity and chromosome instability. AB - Chromosome aberrations are the hallmark of cancer cells. Although a few specific chromosome aberrations are frequently detected in some types of cancer, the majority of karyotypic abnormalities tend to differ between different histological types and between individuals with the same type of cancer. Recent work indicates that telomeres may be directly involved in shaping the karyotypes of tumor cells. In particular, the heterogeneity of telomere lengths within cells may have direct influence on the frequency with which chromosomes engage in telomeric fusions and in subsequent breakage-fusion-bridge cycles. Since telomere length distribution among chromosome arms is a polymorphic trait, difference in distributions between individuals may account, at least in part, for the karyotypic differences found among tumors of the same type. Conversely, if single telomere lengths happen to be inherited, the segregation of particularly short telomeres in families may increase the incidence of specific chromosome aberrations during tumor evolution, and perhaps contribute, along with other factors, to cancer pre-disposition. PMID- 15341023 TI - Polymorphisms of estrogen-metabolizing genes and risk of hepatocellular carcinoma in Taiwan females. AB - Estrogen has been related to the development of hepatocellular carcinoma (HCC). In this molecular epidemiological study, we used logistic regression to compare the genotype frequencies of estrogen-metabolizing genes that are involved in estrogen biogenesis (CYP17), hydroxylation (CYP1A1) and inactivation of the reactive metabolites (catechol-O-methyltransferase, COMT) in HCC patients and control subjects, and determined their relationship with the risk of female HCC. The heterozygous or homozygous variants of high activity CYP17 (A2), high inducibility CYP1A1(m1), and low activity COMT (L) alleles were considered as high-risk genotypes. We found that the risk of HCC was elevated in women harboring either heterozygous or homozygous variants of the CYP1A1 gene and the respective OR (and 95% confidence interval) were 6.61 (1.35, 32.43) and 12.00 (1.73, 83.46). Moreover, we found that the risk of HCC was increased in the female subjects harboring higher numbers of high-risk genotypes, but not in male subjects. The OR for female HCC associated with two putative high-risk genotypes was 12.63 (1.50, 106.37), and the OR for three putative high-risk genotypes was 16.67 (1.82, 152.77). These findings strongly suggest that estrogen play a critical role in female hepatocarcinogenesis. PMID- 15341024 TI - Revised guidance to help prevent kernicterus. PMID- 15341025 TI - Effect of ionizing radiation on gene expression in CD4+ T lymphocytes and in Jurkat cells: unraveling novel pathways in radiation response. AB - To better understand at the molecular level the effect of ionizing radiation in leukocytes, the global transcriptional response to X-ray irradiation was studied in human CD4+ T lymphocytes and in Jurkat cells. Microarray analysis performed on freshly isolated human CD4+ lymphocytes 8 h after an LD50 irradiation dose of 1 Gy revealed that out of 13,825 genes, 1084 were modulated more than 1.5-fold. The most strongly up-regulated genes were predominantly p53 targets. In contrast, exposure of the CD4+ T lymphocyte-derived Jurkat leukemic cell line (with no functional p53 gene) to an equivalent LD50 dose (0.5 Gy) induced a partly different and more limited set of genes. Interestingly, this set of genes belonged to the Rho and cytokine signaling pathways regulated by low-dose ionizing radiation. PMID- 15341026 TI - Neuropsychologic changes and inhaled corticosteroids. PMID- 15341027 TI - Interpretation of isolated positive skin test results for mesquite tree pollen. PMID- 15341028 TI - Latex allergens are not detectable in reconstituted smallpox vaccine or vaccine vial stoppers. PMID- 15341029 TI - Chronic idiopathic urticaria exacerbated with progesterone therapy treated with novel desensitization protocol. PMID- 15341030 TI - Body burden of mercury is associated with acute atopic eczema and total IgE in children from southern Germany. PMID- 15341031 TI - Is glucosamine safe in patients with seafood allergy? PMID- 15341032 TI - Motor and cognitive function evaluation following experimental traumatic brain injury. AB - Traumatic brain injury (TBI) in humans may cause extensive sensorimotor and cognitive dysfunction. As a result, many TBI researchers are beginning to assess behavioral correlates of histologically determined damage in animal models. Although this is an important step in TBI research, there is a need for standardization between laboratories. The ability to reliably test treatments across laboratories and multiple injury models will close the gap between treatment success in the lab and success in the clinic. The goal of this review is to describe and evaluate the tests employed to assess functional outcome after TBI and to overview aspects of cognitive, sensory, and motor function that may be suitable targets for therapeutic intervention. PMID- 15341033 TI - Feminist discourse on sex screening and selective abortion of female foetuses. AB - Although a preference for sons is reportedly a universal phenomenon, in some Asian societies daughters are considered financial and cultural liabilities. Increasing availability of ultrasonography and amniocentesis has led to widespread gender screening and selective abortion of normal female foetuses in many countries, including India. Feminists have taken widely divergent positions on the morality of this practice. Feminists from India have strongly opposed it, considering it as a further disenfranchisement of females in their patriarchal society, and have agitated successfully for legislative prohibitions. Libertarian feminists on the other hand, primarily from the United States, have argued that any prohibition of the use of this technology is a curtailment of a woman's reproductive choices and a violation of her right to make autonomous decisions regarding procreation. Using India as an illustrative case, this paper argues that in the context of what prevails in some societies, an ethical argument that hinges on the principle of autonomy as understood in the West can be problematic. Furthermore, a liberal theoretical assumption that it is always better to have more rather than fewer choices may not hold up well against the realities of life for such women. Although feminists have little disagreement concerning substantive matters, it is in the area of strategy that differences of opinion have arisen, their moral reasoning and responses shaped by the culture, ethnicity, class and race to which they belong. A view that a single 'orthodox' feminism of any variety can embody the aspiration of all women reverts to the problematic issues in the evolution of the rationalistic, individualistic, 'male' ethics against which women have consistently raised objections. PMID- 15341034 TI - Meta-analysis and the science of schizophrenia: variant evidence or evidence of variants? AB - Quantification (meta-analysis) of the neuroscience evidence on schizophrenia shows very modest average differences between patient and control distributions across a great variety of measures and literatures. The strongest findings involve cognitive and psychophysiological measures. Several possible explanations for this situation are reviewed including technical immaturity, methodological variability, dimensional and multiple illness models and the nature of cognitive measurement. An argument is developed that biological subtypes and endophenotypes within the broad diagnostic category of schizophrenia underpin the meta-analytic evidence. Considerations in the use of this evidence to identify illness variants are described and four candidate subtypes are proposed. Schizophrenia is a disease that will resist biological definition until its variants are isolated and extracted from the generic patient population. PMID- 15341035 TI - The problem of abortion: essentially contested concepts and moral autonomy. AB - When one thinks about the ethics of abortion, one inevitably thinks about rights, since it is in terms of the concept of rights that much of the debate has been conducted. This is true of overtly feminist as well as non-feminist accounts. Indeed, some early feminist writers--Judith Jarvis Thomson and Mary Ann Warren, for example--employ a model of rights that is indistinguishable, or virtually indistinguishable, from that of their non-feminist counterparts. However, more recent feminist writers have developed a different understanding of 'a woman's right to choose.' In this paper, I will begin by outlining the non-feminist debate over the moral permissibility of abortion. I will suggest that this debate is irresolvable, since at its heart is an 'essentially contested concept', that of personhood. I will then consider the way in which some feminist writers have attempted to reconceive the terms of the abortion debate and suggest an expanded account of women's right to abortion, drawing on the work of Susan Sherwin. Finally, I will argue that there is a further element to a 'woman's right to choose' that expands on and provides a conceptual link between feminist and non feminist understanding of abortion. PMID- 15341036 TI - Adequate conscious life and age-related need: F.M. Kamm's approach to patient selection. AB - Kamm's approach to patient selection qualifies the notion that fairness makes need for scarce, transplantable organs inversely proportional to age. She defines need as how much adequate conscious life a person will have had before death. Length of adequate conscious life correlates highly with age. If so, then younger persons are usually needier than older ones. Since Kamm allows for past periods of non-adequate conscious life, I argue that this correlation may be neither as close, nor as easy to apply, as she wants it to be. Fairness should require assessment of experiential content in determining how long one's life has been adequately conscious. I argue that such assessments involve some of the goods of experience and quality of life judgements that Kamm thinks a reliance on adequate conscious life will avoid. PMID- 15341037 TI - Interactions between the cardiovascular and pain regulatory systems: an updated review of mechanisms and possible alterations in chronic pain. AB - Endogenous pain regulatory system dysfunction appears to play a role in the maintenance of chronic pain. An important component of the pain regulatory process is the functional interaction between the cardiovascular and pain regulatory systems, which results in an association between elevated resting blood pressure (BP) and diminished acute pain sensitivity. This BP/pain sensitivity relationship is proposed to reflect a homeostatic feedback loop helping restore arousal levels in the presence of painful stimuli. Evidence is emerging that this normally adaptive BP/pain sensitivity relationship is significantly altered in chronic pain conditions, affecting responsiveness to both acute and chronic pain stimuli. Several mechanisms that may underlie this adaptive relationship in healthy individuals are overviewed, including endogenous opioid, noradrenergic, and baroreceptor-related mechanisms. Theoretical models are presented regarding how chronic pain-related alterations in the mechanisms above and increased pain facilatory system activity (central sensitization) may contribute to altered BP/pain sensitivity interactions in chronic pain. Clinical implications are discussed. PMID- 15341038 TI - The pro-life argument from substantial identity: a defence. AB - This article defends the following argument: what makes you and I valuable so that it is wrong to kill us now is what we are (essentially). But we are essentially physical organisms, who, embryology reveals, came to be at conception/fertilisation. I reply to the objection to this argument (as found in Dean Stretton, Judith Thomson, and Jeffrey Reiman), which holds that we came to be at one time, but became valuable as a subject of rights only some time later, in virtue of an acquired characteristic. I argue against this position that the difference between a basic, natural capacity and some degree of development of such a capacity is a mere difference in degree, that this position logically implies the denial of equal personal dignity, and that the selection of the required degree of development of a capacity is necessarily arbitrary. PMID- 15341039 TI - Essential properties and the right to life: a response to Lee. AB - In 'The Pro-Life Argument from Substantial Identity: A Defence', Patrick Lee argues that the right to life is an essential property of those that possess it. On his view, the right arises from one's 'basic' or 'natural' capacity for higher mental functions: since human organisms have this capacity essentially, they have a right to life essentially. Lee criticises an alternative view, on which the right to life arises from one's 'developed' capacity for higher mental functions (or development of some other accidental property). I argue that his criticisms of this alternative view are misguided or self-defeating, and that there are good reasons to hold we have a right to life accidentally rather than essentially. PMID- 15341040 TI - To strengthen consensus, consult the stakeholders. AB - CIOMS has been criticised for not adequately consulting stakeholders about its revised ethical guidelines regarding medical research. Political and logistical issues that arise in democratic processes and open exchange of information probably contributed to this exclusion. What might CIOMS have done to be more inclusive and attain broader consensus on its proposed revisions? Consensus is dynamic, and evolves as a community digests new information and perspectives. Engaging the public (and particularly the stakeholders) in discussion about the revisions would have generated broader consensus. It would have encouraged more stakeholders (i.e. researchers, potential research participants, research institutions, or governments) to buy in. CIOMS needs a process to encourage dialogue and stakeholder input. The CIOMS guidelines themselves promote stakeholder consultation and capacity building, but CIOMS has done relatively little to distribute or promote its own guidelines. CIOMS should do more to promote its revised guidelines, and engage stakeholders in dialogue. This paper explores the bioethics debate about universal and relative values to illustrate the value of consultation and consensus building. It concludes that like research sponsors, CIOMS and similar organisations have an ethical responsibility to facilitate capacity building in less developed areas, and to participate more actively in consensus building. PMID- 15341041 TI - The quest for legitimacy: comment on Cox Macpherson's 'To strengthen consensus, consult the stakeholders'. PMID- 15341042 TI - Regenerative medicine: stem cells and the science of monstrosity. AB - The nineteenth century science of teratology concerned itself with the study of malformations or "monstrosities", as they were then called. The first major contribution to the field was the work of Isidore Geoffrey Saint-Hilaire, Historie Generale et Particuliere des Anomalies de l'Organisation chez l'Homme et les Animaux, published in 1832, whose classifications formed the basis for the later experimental science of teratogeny, the art of reproducing monstrosities in animal embryos. In this article, I will argue that recent developments in the field of regenerative medicine can be situated in the tradition of teratological and teratogenic studies dating back to the nineteenth century. In particular, I will be interested in the historical link between studies in teratogenesis (the artificial production of teratomas) and stem cell research. Recent advances in stem cell research, I will suggest, return us to the questions that animated nineteenth century investigations into the nature of the monstrous or the anomalous. In the process, our most intuitive conceptions of "life itself" are undergoing a profound transformation. PMID- 15341043 TI - Can Mary Shelley's Frankenstein be read as an early research ethics text? AB - The current popular view of the novel Frankenstein is that it describes the horrors consequent upon scientific experimentation; the pursuit of science leading inevitably to tragedy. In reality the importance of the book is far from this. Although the evil and tragedy resulting from one medical experiment are its theme, a critical and fair reading finds a more balanced view that includes science's potential to improve the human condition and reasons why such an experiment went awry. The author argues that Frankenstein is an early and balanced text on the ethics of research upon human subjects and that it provides insights that are as valid today as when the novel was written. As a narrative it provides a gripping story that merits careful analysis by those involved in medical research and its ethical review, and it is more enjoyable than many current textbooks! To support this thesis, the author will place the book in historical, scientific context, analyse it for lessons relevant to those involved in research ethics today, and then draw conclusions. PMID- 15341044 TI - Ethics in population-based genetic research. AB - Population-based genetic research, including genetic epidemiology, shows tremendous potential to elucidate the role of genes as causal factors in complex and common human diseases. Like all research with human subjects, full realization of these benefits requires careful attention to its ethical conduct, establishing an appropriate balance between individual protections and the advancement of scientific and medical knowledge. This article reviews the growing literature on genetics research and ethics to describe some of the fundamental ethical issues in population-based genetics research, including research design, recruitment and informed consent, and dealing with research results. Its focus is on areas where consensus is forming and where future work is needed. PMID- 15341045 TI - Ethics in behavioral genetics research. AB - As research in behavioral genetics uncovers the genetic contribution to human behavior, it will undoubtedly further our understanding of normal human variation in many behavioral traits, such as personality, intelligence, and sexuality. This research also shows great potential for the diagnosis, treatment, and prevention of mental illnesses. Recent findings underscore this potential and document the increasing validity of research methods--methods that in the past have led to mistaken inferences about genes "for" violent behavior and homosexuality. Although all research with human subjects requires adequate attention to its ethical aspects, certain ethical issues involved with behavioral genetics are particularly acute and deserve careful attention. This article reviews these selected major ethical issues arising in (1) the conduct of behavioral genetics research; and (2) the application of its research findings. While some of the ethical concerns in the latter category are likely to be of substantial importance and animate considerable popular concern, they currently fall outside the realm of traditional research review. Determining how to deal with these concerns should be a focus of future scholarly work. PMID- 15341046 TI - Ethical challenges in international HIV prevention research. AB - Members of an HIV-prevention research network were asked to describe ethical challenges faced in their work. Major themes included acceptable standards of care for participants, defining research of relevance to host countries, reducing risks related to stigma, designing research that meets local needs without contributing to an inadequate status quo, and ensuring informed consent for complex research with potentially vulnerable participants. The challenges are interrelated and highlight the need for innovative, practical strategies to be incorporated into the planning, design, and conduct of HIV prevention trials. Research in applied ethics to support decision-making about HIV prevention research is needed, along with ethics training, mechanisms to support community wide benefit from research, and expanded dialog on ethics surrounding HIV prevention and public health research. PMID- 15341048 TI - Play it safe. Considering reuse? A solid reprocessing contract is your safety net. PMID- 15341047 TI - Physicians' understanding of consent requirements for phase I clinical trials in cognitively impaired or highly vulnerable populations. AB - We investigated physicians' attitudes about entering patients who cannot give informed consent or who are of a vulnerable population into clinical trials. A survey instrument asked a nationwide sample of practicing physicians about whether ten hypothetical patients could be enrolled in a phase I clinical trials. The impact of demographic variables on the number of scenarios viewed as completely or somewhat acceptable was analyzed via student's T tests or analysis of variance (ANOVA) as applicable. All significant (p<0.01) variables were entered into a multiple logistic regression model. Eighty-four percent of respondents indicated that at least one case scenario was acceptable. A majority of those who conduct clinical trials (62%), who had training in the ethics of clinical research (78%), and who sit on institutional review boards (IRBs) (83%) approved of at least one case scenario. Physicians approved of the entry of some patients who cannot give informed consent or who are of a vulnerable population into clinical trials. More effective education on the guidelines involving clinical research should be available to practicing physicians, residents, and medical students. There should also be assurance that physicians who conduct clinical trials or who sit on IRBs have the requisite knowledge about the ethics of clinical research. PMID- 15341049 TI - The devil's in the details--so focus on the fine print! PMID- 15341050 TI - Who's minding the store? PMID- 15341051 TI - Fattening of America spurs demand for special furniture. PMID- 15341052 TI - Dispense-a-scrub. Vending machines, new policy slash high scrub use. PMID- 15341053 TI - Talk your way to success. PMID- 15341054 TI - TNA's 2001 leglslative agenda. Shortage, workplace and more. PMID- 15341057 TI - [Does the catalepsy phenomenon reflect the functional dopaminergic activity in pharmacological investigations?]. AB - The dynamics of spontaneous and haloperidol-induced catalepsy in mice has been studied. It is established that the degree of manifestation of the spontaneous catalepsy is directly related to the number of manipulations (mice standings in the "lector position"). At the same time, the intensity of catalepsy (both spontaneous and haloperidol-induced) was not related to the mice response to apomorphine injections. It is concluded that spontaneous catalepsy rather insignificantly influences the results of pharmacological tests, but this factor has to be taken into account in the study of physiological mechanisms. The absence of correlations between catalepsy manifestations and the apomorphine test results is indicative of complexity of the mechanism of this disorder and cannot be attributed entirely to the violation of dopaminergic processes in CNS. PMID- 15341058 TI - [The effect of ladasten on the activity of cAMP-dependent protein kinases and the phosphorylation of proteins in brain cells of rats]. AB - The effect of ladasten (50 mg/kg) on the activity of cAMP-dependent protein kinases (represented by protein kinase A (PKA)) in the soluble protein fraction of rat brain was studied. Changes in the PKA activity were monitored in the initial period of drug action (1-12 h). In this stage, ladasten increased the PKA activity, and the level of phosphorylation of one isoform of the main protein myelin (14 kD) and of the proteolipid protein (30 kD). It is concluded that the pharmacological activity of ladasten is related to both cAMP-dependent and cAMP independent signaling systems. PMID- 15341059 TI - [The effect of K(ATP)-channel activation on the electrical stability of myocardium in rats with postinfarction cardiosclerosis]. AB - Opening of the ATP-dependent K-channels (K(ATP) channels) upon intravenous administration of the cardioselective activator BMS 180448 (3 mg/kg) decreased the ventricular fibrillation threshold (VFT) in rats with postinfarction cardiosclerosis (PIC). Preliminary injection of the selective K(ATP) channel blocker glibenclamide (0.3 mg/kg, i.v.) completely abolished the profibrillatory effect of BMS 180448. At the same time, the mitochondrial K(ATP) channel blocker 5-hydroxydecanoic acid (5 mg/kg) did not influence the proarrhythmogen activity of BMS 180448. Simultaneous administration of the sarcoK(ATP) channel inhibitor HMR 1098 (3 mg/kg) and BMS 180448 increased the VFT up to a level in intact animals. Administration of the mitoK(ATP) channel activator diazoxide (5 mg/kg) after preliminary treatment with guanethidine (50 mg/kg) increased the VFT in rats with PIC. It is concluded that opening of the mitoK(ATP) channels increases the cardiac electrical stability in rats with PIC. PMID- 15341060 TI - [Effect of hydroxymethylethylpyridine succinate on the cardiac electrophysiological and hemodynamic parameters during experimental thoracotomy and acute myocardial ischemia]. AB - Effects of the natural antioxidant hydroxymethylethylpyridine succinate (mexidol) on the electrophysiological and hemodynamic cardiac parameters were studied on narcotized mongrel cats with experimental thoracotomy and acute myocardial ischemia, as manifested by sinoatrial node automatism, conduction, effective refractory period, and excitability of myocardium. The rate of contractility (dp/dtmax) in the left ventricle and the arterial pressure level (p) were monitored using a special differentiating device. The antiarrhythmic activity of mexidol was studied by ECG. The cardioprotective effect of the drug is manifested by normalization of the electrophysiological parameters during operation (accompanied by artificial lung ventilation, thoraco- and pericardiotomy). The positive effect was also observed in operated animals with acute ischemia model. The prophylactic administration of mexidol in a dose of 7 mg/kg prevented the cardio-depressant consequences of acute ischemia and reduced the risk of ventricular arrhythmia during coronary artery occlusion. PMID- 15341061 TI - [Studying the mechanism of antiarrhythmic action of a tertiary derivative of lidocaine]. AB - The investigation of electrophysiological parameters of the cardiac activity in cats showed that, a derivative of lidocaine with glutamic acid (anion), does not influence the cardiac pacemaker and the myocardial excitability of atrium and ventricles, but decreases the atrioventricular conduction and increases the refractory period of myocardium in the left ventricle of intact heart. Under the conditions of myocardial ischemia, LKhT-3-00 exhibits a negative chronotropic effect. Lidocaine glutamate increases the refractory period of atrium and decreases the excitability and refractory period of ventricles. PMID- 15341062 TI - [Hemodynamic and rheological effects of polyetox in rats with crush syndrome]. AB - Polyetox, a medicinal form of high-molecular-weight poly(ethylene oxide) (HMWPEO) improved peripheral blood supply, normalized the overall oxygen consumption, decreased erythrocyte aggregation, and reduced blood viscosity at low shear rate, and restored the antiturbulent properties (hydrodynamic index) of blood in the experiments on rats with crush syndrome. In rats with low resistance, polyetox increased the cardiac output. PMID- 15341063 TI - [Effect of aminophylline on fatigue of inspiratory muscles]. AB - Effect of aminophylline on the fatigue of inspiratory muscles and ventilation disorders induced by respiratory resistive load was studied in cats. The administration of aminophylline on the background of inspiratory fatigue increases transdiaphragm pressure, restores bioelectric activity of inspiratory muscles and phrenic nerve, and decreases the respiratory duty cycle and rate. It is concluded that aminophylline in a dose of 24 mg/kg (i.v.) is capable of compensating the inspiratory muscle fatigue via both central and peripheral mechanisms, the latter effect being probably of greater importance. PMID- 15341064 TI - [Antiulcer activity of furoxanopyrimidine derivatives]. AB - A series of four new furoxanopyrimidine derivatives was synthesized and studied with respect to antiulcerous, antisecretory, and antibacterial activity. Two compounds exhibit antiulcerous effect not accompanied (in contrast to the well known H2 receptor blockers, quiditene, and other antiulcerous drugs) by inhibition of gastric acid secretion. No one of the studied compounds exhibited antibacterial activity in the tests with Helicobacter pylori. PMID- 15341065 TI - [The effect of bromantan on the sexual behavior and conception in rats]. AB - The results of experiments on rats showed that the psychotropic drug bromantan affects the sexual behavior and pairing activity. A three-day treatment with bromantan (30 and 300 mg/kg, p.o.) predominantly increased the sexual proceptivity, whereas a chronic administration (for 2 months in males and 2 weeks in females) produced a dose-dependent increase in both proceptivity and receptivity. The observed effects are probably related to the dopamine-positive action of bromantan. PMID- 15341066 TI - [A comparative study of the antioxidant activity of salbutamol hemisuccinate and essentiale in rats with experimental phetoplacental insufficiency]. AB - The experiments on rats with phetoplacental insufficiency induced by tetrachloromethane showed that beta2-mimetic salbutamol hemisuccinate (salgim) exhibits antioxidant activity. The effect was most pronounced in placenta, where salgim exceeded in activity the well-known reference drug essentiale. PMID- 15341067 TI - [Comparative assessment of cell mechanisms of the antidiabetic action of a new organic derivative of oxovanadium(IV) and vanadyl sulfate]. AB - The antidiabetic effect of a new organic derivative of vanadium(IV) oxide with isonicotinic acid hydrazide (compound no. 8), as manifested by the action upon the alpha and beta cell populations in Langerhans islands of the pancreas, was studied in rats with alloxane diabetes model in comparison to the analogous effect of the inorganic drug vanadyl sulfate. The hypoglycemic activity of compound no. 8 was comparable with that of vanadyl sulfate. The results of immunohistochemical and morphometric investigation showed that both preparations produced a reliable increase in the population of insulin-producing cells and a decrease in the (alloxane-enhanced) population of alpha-cells in the pancreatic islands. PMID- 15341068 TI - [Effect of the joint administration of ketamine and propofol on the lipid metabolism and peroxidation in rats]. AB - Ketamine (50 mg/kg, i.p.) increases the intensity of lipolysis and lipid peroxidation (LPO) in the hepatic and cardiac tissues of Wistar male rats, as manifested by an increase in the content of nonetherified fatty acids and cholesterol in the blood serum and in the content of LPO products in the blood serum and in the liver and heart tissues. Propofol in the same dose also influences the lipid metabolism and LPO intensity, but to a lower extent. The joint administration of both ketamine (25 mg/kg, i.p.) and propofol (25 mg/kg, i.p.) leads to averaging of some characteristics of the lipid metabolism and LPO. PMID- 15341069 TI - [Changes in the immune reaction of animals under conditions of drug-induced activation and blockade of D1 dopamine receptors]. AB - A highly selective dopamine D1 receptor agonist (compound SKF 38393) administered in a dose of 20 mg/kg produced a significant increase in the number of rosette forming cells (RFCs) in the spleen of Wistar rats on the 5th day upon their immunization with goat erythrocytes (5 x 10(8)). At the same time, a specific blocker of these receptors (compound SCH 23390) in a dose of 0.5 or 1 mg/kg inhibited the immune response of rats. In C57BL/6J mice, SCH 23390 (0.25, 0.5, or 1 mg/kg) also significantly reduced the number of RFCs. Preliminary blockade of the D1 dopamine receptors with SCH 23390 (1 mg/kg) prevented the immunostimulating effect of SKF 38393. These data indicate that D1 dopamine receptors are involved in immunomodulation. PMID- 15341070 TI - [Immunometabolic effects of interactions involving fat- and water-soluble vitamins in toxic forms of anemia]. AB - Phenylhydrazine or sodium nitrite injections reduce the functional metabolic activity of neutrophils (leukocytes) in the peripheral blood, suppress the development of humoral immune response and delayed type hypersensitivity, decrease the energetic potential and the antioxidant status of erythrocytes, and induce the appearance of immunosuppressive activity in these cells. Retinol acetate, tocopherol acetate, and phylloquinone administered in various pair combinations reduce the degree of changes in the parameters of nonspecific resistance and immunological reactivity. Pyridoxine and cobalamin increase the immunomodulating activity of erythrocytes modified by phylloquinone. PMID- 15341071 TI - [The functional state of thymus cells under prolonged combined action of phenobarbital and E. coli lipopolysaccharide]. AB - The effect of separate and combined 10-day treatment with phenobarbital (PB) in a single daily dose of 35 mg/kg and E. coli lipopolysaccharide (LPS) in a dose of 25 microg/kg was studied in white rats. The results were evaluated by the rate of DNA synthesis in the culture of T mitogen stimulated thymocytes, the activity of glucose-6-phosphate dehydrogenase (G-6-PDG) in the thymus, and the activity of acetylcholine esterase (ACE) and the content of catecholamines (CA) in the nerve fibers of this organ. It was established that the 10-day treatment with PB inhibits the proliferative activity in concanavalin A stimulated lymphocytes of rat thymus, decreases the ACE activity, and increases the CA content in the nerve fibers. The repeated injections of E. coli LPS on the background of PB led to more pronounced negative changes in the functional activity of lymphoid cells in the thymus, manifested by a decrease in the G-6-PDG activity and the rate of DNA synthesis in the thymus and by an increase in the level of biogenic amines in the nervous fibers of this organ. PMID- 15341072 TI - [Determination of nonmetabolized antipyrine in human urine]. AB - A method for evaluating the renal clearance of antipyrine is proposed. The analyses were performed for a group of healthy females of reproductive age. The renal clearance of nontransformed antipyrine is found to be significant and the basic indices of renal activity (diuresis, creatinine excretion) are correlated with the drug concentration in saliva. A single administration of antipyrine in a dose of 10 mg/kg does not influence the renal function of healthy volunteers. The proposed photometric method of antipyrine determination in urine (under the conditions of loading with 0.5% NaCl solution at an amount of 0.5% of body weight) is sufficiently sensitive and provides information about the renal clearance of antipyrine, offering an important supplement to the data obtained by conventional methods. PMID- 15341073 TI - [Functional-behavioral profile of new cyclic GABA analogs in acute toxicity tests]. AB - The properties of karphedone and phepyrone--new phenyl derivatives of pyrrolidone possessing nootropic activity--were studied in the course of the acute toxicity tests on rats. The drug effects were evaluated in terms of their integral influence on the state and behavior of test animals. The real therapeutic range and the profit/risk ratio of karphedone were comparable with those of the reference drug pyracetam and exceeded by a factor of 1.3 the corresponding values for phepyrone (irrespective of the LD50 values). The results give grounds for the further preclinical investigation of karphedone. PMID- 15341074 TI - [The effects of cytamines and their combinations with ecdystene, apilak, vitamax, and essentiale on the work capability of athletes]. AB - Cytamines (hepatamin, epifamin, and suprenamin) administered in various combinations increase the working ability of high-rank athletes in experiments. These cytamines also potentiate the effects of apilak, ecdystene, vitamax, and essentiale preparations on the exercise tolerance. PMID- 15341075 TI - [The antioxidant activity of S-(4-quinazolyl)mercaptoacetic acid benzylidene hydrazides in vitro in the "nitrosation stress" model]. AB - The "nitrozation stress" causes modification of the antioxidant enzymes leading to a decrease in their activity. A decrease in this activity and the resulting increase in the production of OH* and NO2* radicals via ONOO* decomposition leads to a significant increase in the extent of free-radical peroxidation during the "nitrozation stress". These factors are unfavorable from the standpoint of oxidative phosphorylation, which is manifested by inhibited activity of the oxidative enzymes. S-(4-Quinazolyl)mercaptoacetic acid benzylidene hydrazides exhibit a high antiradical activity related to the presence of a mercapto group in the quinazoline fragment. The activity depends on the position of this group on and the number of substituents in the benzylidene fragment. The compounds studied reduce the extent of damage in the brain homogenates of rats under conditions of nitrozation stress. This is manifested by inhibition of the free radical peroxidation and an increase in the activity of antioxidant and prooxidant enzymes. PMID- 15341076 TI - [Wound-healing properties of solid immunobiological medicinal forms for local application]. AB - The results of experiments on rats showed a high healing potential of three collagen-based coating materials containing a combination of biologically active substances including human leukocyte interferon, alfalfa extract, and eubiotic bactisporin. The application of new materials significantly accelerated the healing of model wounds. PMID- 15341077 TI - [Antisense oligonucleotides: a new class of biologically active molecules]. AB - Data available in literature on the biological activity of antisense oligonucleotides are reviewed with emphasis on the results of tests of the related antitumor and antiasthmatic preparations and the basic novelty of these drugs. PMID- 15341078 TI - [The role of support afferents in organisation of the tonic muscle system]. AB - Results of experimental studies are reviewed that point out to the leading role of the support afferents in control of structural-functional properties of the tonic muscle system. It is shown that the support afferents play a role of the trigger in the postural system, the trigger enhancing (when the support is present) or inhibiting (when the support is withdrawn) the activity of tonic motor units (MU's). Under the absence of support condition, recorded in extensors are: an obvious decline of the muscle stiffness and the maximal voluntary force; a significant decrease of the absolute force of the isometric contraction of single skin muscle fibers evoked by Ca++; a prominent decline of the tonic muscle fibers transversal size; and the transformation of the myosin phenotype from slow to fast one. Mechanical stimulation of the support zones of soles in the regimes of locomotion (slow and fast stepping) under the absence of support condition eliminates all the above effects. PMID- 15341079 TI - [Endothelial microtubules as integral part of the mechanism controlling the level of stimulated secretion of van Willebrand factor]. AB - We used double immunofluorescence and electron microscopy to study the spatial relationships between Weibel--Palade bodies (WPBs) and cytoskeletal elements in endothelial cells treated with thrombin or cytoskeleton-damaging agents. We have found that some WPBs undergo translocation towards the centrosome in 5 min in the cells treated with thrombin, cytochalasin B or calyculin A. The cells treated with thrombin or cytochalasin exhibit depletion of WPBs, whereas WPBs found at the cell periphery were colocalized with intermediate filaments. There was a precise colocalization observed between the WPBs and microtubules in the calyculin-treated cells in which all WPBs undergo centrosome-directed translocation within 15 min after the agent addition. When vimentin filaments were induced to collapse by demecolcine, intermediate filaments and WPBs both translocated to the perinuclear region. The data provide the first direct evidence that secretory granules utilize microtubules to move in retrograde direction, i.e., away from the plasma membrane, towards the centrosome. We suggest that anterograde movement of WPBs is dependent on their interaction with vimentin filaments. PMID- 15341080 TI - [The role of stretching the sinus node artery in initiation of atrial fibrillation]. AB - In anaesthetised dogs with open chest (n = 10), against the background of the Gd3+ (a blocking agent of mechano-sensitive ion channels), acetylcholine perfusion induced no significant changes either in probability of atrial fibrillation occurrence or in the paroxysm duration. The Gd3+ did not alter the deceleration of the sinus rhythm either. Therefore the mechanism of spontaneous occurrence of atrial cholinergic fibrillation seems not to be associated with the trigger activity induced by an increased blood pressure in the right atrium. PMID- 15341081 TI - [Plasminogen activator of urokinase-type: mechanisms of involvement in vessel remodeling and angiogenesis, gene therapy approaches to ischemia]. AB - The review summarizes data obtained by the authors and other laboratories concerning the role of urokinase plasminogen activator in vessel remodeling and angiogenesis. The data have shown that urokinase is involved in unfavorable vascular remodeling during the development of restenosis, atherosclerosis and also in the regulation of angiogenesis. Urokinase is a promising target for therapeutic interventions aimed at restenosis prevention. Urokinase gene therapy may be a perspective strategy for the treatment of tissue ischemia. PMID- 15341082 TI - [Photodynamic approaches to elimination and prevention of atherosclerotic changes in the vessels]. AB - Photodynamic therapy (PDT) is a method based on a selective accumulation of a photo-sensitive dye in the target cells with their subsequent destruction with the aid of a low-intensity laser radiation of a certain wavelength. It is widely used today in clinical practice, mostly in oncology. This review elucidates possibilities of using the PDT in prophylactics and treatment of cardiovascular pathological conditions, atherosclerosis in particular. Data are cited indicating a successful application of the PDT for preventing the intima hyperplasia following and angioplasty. A possibility of using the PDT for treatment and prophylactics of atherosclerotic alterations in the vessels with due consideration of specifics of formation and cullular composition of the atherosclerotic lesions. PMID- 15341083 TI - [A new actin-binding area of the myosin light chains' high-molecular kinase]. AB - High-molecular weight isoform of myosin light chain kinase (MLCK-210) contains a unique N-terminal domain not present in a low-molecular weight isoform, MLCK-108. Functional features of this domain are poorly studied. We provide evidence that N terminal area of the unique MLCK-210 domain incorporates previously unrecognized actin-binding region that could be involved in the interaction of MLCK-210 with the actin cytoskeleton. PMID- 15341084 TI - [Antiplatelet effects of glycoproteins IIb-IIIa antagonist monafram]. AB - First Russian glycoprotein (GP) IIb-IIIa antagonist, preparation Monafram, is the F(ab')2 fragment of anti-GP IIb-IIIa monoclonal antibody FRaMon. In in vitro experiments it was shown that Monafram blocked platelet aggregation induced by ADP and thrombin; reduced secretion from platelet granules; and due to simultaneous interaction with two GP IIb-IIIa molecules almost irreversibly bound to platelet surface. Monafram clinical trials were performed in healthy volunteers (n = 10) and in patients with ischemic heart disease undergoing high risk coronary angioplasty (n = 153). Monafram intravenous bolus administration at 0.25 mg/kg decreased ADP-induced platelet aggregation by more than 90, 80, 60 and 30% at 1, 12, 24 and 72 h after injection, respectively. No significant differences were detected between antiaggregatory effects of Monafram and ReoPro introduced at 0.25 mg/kg bolus + 12 h infusion at 0.125 microg/kg per min. Durable inhibition of aggregation after Monafram administration was mediated by platelet-bound preparation--free Monafram was cleared from plasma within 12 h, while platelet-bound preparation occupied more than 90, 70-80 and 40-50% of GP IIb-IIIa at 1, 12-24 and 72 h after injection, respectively. Major bleedings and allergic reactions were detected in none of patients, deep thrombocytopenia--in one patient and antibodies against Monafram--in 5% of patients. Within one month after coronary angioplasty Monafram decreased the number of end points (fatal and nonfatal myocardial infarction and angina recurrence) from 11.4 to 3.3%. PMID- 15341085 TI - [The role of the central nervous system imidazoline receptors in cardiovascular manifestations of emotional stress]. AB - The effect of central nervous system imidazoline receptors activation on basal blood pressure level, heart rate and arterial baroreceptor reflex in steady state and aversive emotional tension was tested in experiments on alert WKY, SHR and white bastard rats. It was found that the brain imidazoline receptors activation led to arterial baroreceptor reflex rise (both in resting and in emotional tension) and caused an emotional stress pressor effects decrease. No data proving involvement of imidazoline receptors in functioning of the systems maintaining level of blood pressure, were found. PMID- 15341086 TI - [Computer cardiokymography. On its way to long-term noninvasive monitoring of cardiac performance in daly life]. AB - A functioning heart generates low-frequency and low-amplitude movements (up to 40 (mrn) of soft tissues in the percardial area. Since the 1950-s of the XX-th century cardiologists have studied these movements to employ them in the diagnostics of ischemic heart disease (IHD). Indeed, changes of the movements have found their diagnostic application in this field. If blood supply to a certain area of ventricular myocardium is insufficient the contractions in this area diminish and even ceases. After systolic increase in ventricular pressure this area dilates and forces intercostal tissues out, causing a "bulge" wave on the record. For recording the patient must be in supine position and hold the breath. High frequency electromagnetic field generator was used in an attempt to examine the heart movements. It was found that such instrument measures the thickness of the air-gap that serves as a dielectric or a "capacitor" which forms between the skin and approaching it flat inductive coil. Nevertheless, such an instrument (cardiokymograph, CKG) can reveal changes in contractions of ischemic myocardium and markedly increases the sensitivity of the exercise ECG test. However, CKG can be recorded only after termination of the test. This principal disadvantage has been overcome by software that allowed the recording of CKG during the test. A low-frequency capacitance transducer developed recently by Chinese researchers has provided a simultaneous recording of several CKG. After modifying the transducer, we have established that 1) the shape of CKGs is the same as that CKGs recorded by all our predecessors, 2) CKGs are reproducible even within a 3-month interval, 3) the enormous mechanical interference caused by respiratory movements and, to a greater extent, by the vigorous movements of the legs and trunk during veloergometry can be eliminated by digital processing of the transducer signals, and 4) pathological changes appear in CKG of patients with IHD, which enables one to stop veloergometry at lower loads, thus increasing the safety of the test. Since for large medical insurance companies very simple and inexpensive cardiokymograph are quite unprofitable, their commercially production in USA and in Germany has been stopped. However, the goal of cardiokymography: a real-time, beat-to-beat, long-term monitoring of cardiac function in daily life, remains the major factor determining the future of the method. PMID- 15341087 TI - [The brain as an endocrine gland in adult and in developing ofranism]. AB - It is suggested that the abult mammals' brain is, on the one hand, a conglomerate of neuronal assemblies within which information is being transmitted with the aid of chemical signals, and on the other hand, a considerable part of the neurons are secreting cells and their accumulations by their functional characteristics are identical to endocrine glands. Proceeding from the brain dualism, a question arises: which of the two functions is the pimary one? It seems that, prior to forming specific interneuronal links and blood-brain barrier, the neurons function as secreting cells and the brain--as an endocrine gland. The physiologically active substances (FASs) entering the general circulation system from the brain seem to take part in regulation of development of the visceral target organs and the brain itself. Forming of the interneuronal links and blood brain barrier leads to a qualitative jump in the brain development followed by a considerable limitation of its endocrine functions. According to this concept, the brain, prior to the moment of forming of the interneuronal links and blood brain barrier, does not influence the development of the whole organism whereas development of the brain itself is regulated by the hormones of the foetus and placenta's endocrine glands. PMID- 15341088 TI - [Potential influence of nicotine on inflammation and induction of autoantibodies in rats with adjuvant arthritis]. AB - The influence of chronic administration of nicotine diluted in the drinking water on the parameters of systemic inflammation and autoimmune processes in rats (August line) with adjuvant-induced arthritis, were studied. The experiments have shown that nicotine acts as an antiphlogistic means (the amount of C-reactive protein rises in the blood) and activates autoimmune processes: induction of rheumatic factor, of autoantibodies to serotonin, and glial fibrillar acid protein. It was supposed that nicotine has a potential impact on immunocompetent cells. PMID- 15341089 TI - [Yeast as a model for studying the prion and amyloid occurrence]. AB - Recent data on the use of yeast as a model for studying the molecular basis of prion infection are summarized. In contrast to mammalian prions, which are related to incurable neurodegenerative diseases, yeast prions determine the appearance of non-chromosomally inherited phenotypic traits. Prions of yeast are structurally similar to amyloids of mammals and their replication involves not only growth, but also fragmentation of prion amyloid-like fibrils. In mammals the fragmentation should lead to an increase in infectious titer. The use of yeast for study of the mechanisms of human amyloidoses, development of new anti-prion drugs and search for new proteins with prion properties is described. PMID- 15341090 TI - [The protein arrestin as a regulator of the phototransduction process and as a factor of pathogenesis of the eye diseases]. AB - In the first part of the paper, the results of the investigation of the rhodopsin arrestin interaction are presented. The results were mainly obtained with the technique of the selective labelling of the rhodopsin and arrestin SH-groups and the rhodopsin limited proteolysis. These results are discussed in the frame of the latest data on the three-dimensional structure of arrestin. In the second part of the paper, results of the antigenic properties of arrestin (S-antigen) and its role in the pathogenesis of the retina diseases are summarized. The data on the role of the autoimmune processes in the pathogenesis of diabetic retinopathy are presented. We have also described the results of the use of the elaborated technique of the immune diagnostics in the prognosis of the diabetic retinopathy and retinopathy of the premature babies. PMID- 15341091 TI - [The 5th International Symposium of Vitreoretinal Surgery]. PMID- 15341092 TI - [Neuroprotection and vasoprotection in glaucoma]. PMID- 15341093 TI - [Early primary congenital glaucoma]. AB - Early primary congenital glaucoma belongs to the category of primary congenital glaucomas and is regarded as one of the most frequent form of congenital glaucoma, in which case the intraocular hypertension usually appears in as early as the first year of living before the age of three or four, caused by mesodermic anomalies during the development of anterior chamber angle, leading to the stretching of the scleral wall and to the increase of the eye, known as buphthalmos and which associates with no other ocular or systemic anomaly. It is more frequently met as a bilateral one. The diagnostic methods include: history, the measurement of corneal diameter, the slitlamp examination, tonometry, gonioscopy, ophthalmoscopy, ultrasonography. The basic treatment is a surgical operation and must be provided as soon as possible. PMID- 15341094 TI - [Biochemistry of Chirila's keratophacoprothesis]. PMID- 15341095 TI - [Aspects of optical coherence tomography in diabetic retinopathy]. AB - Optical coherence tomography is a new and very useful technique for diabetic patients. It is used to determine the early changes in diabetic retinopathy and to help the ophthalmologist to choose the appropriate therapy. PMID- 15341096 TI - [Congenital grouped pigmentation of the retina]. AB - A case of grouped pigmentation of the retina with unilateral localisation is presented. The positive diagnosis of this congenital abnormality is decided upon the aspects of the retinal grouped pigmentations and it is confirmed using additional test: angiography, electrophysiological methods, visual field test and adaptometry. PMID- 15341097 TI - [Pseudovitelliform macular degeneration]. AB - We report the case of a 41-year old-male, which complained of mild visual acuity loss at the left eye. His medical and family histories were not significant. The ophthalmoscopic and fluorescein angiographic findings in this man were typical for pseudovitelliform macular degeneration. Aspects of pathogeny, diagnosis and differential diagnosis are discussed. PMID- 15341098 TI - [Retinal tumors of the adult--two case reports]. AB - Most frequently, retinal tumors that affect adults are rare, benign lesions that arise from vascular structures (hemangioma), neuroglial structures (astrocytoma, massive gliosis of the retina) or neuroepithelial cells (occasional forms of retinoblastoma that spontaneously stop growing and do not achieve or lose their malignant character). Retinal metastasis might be encountered but the settling of cancer cells at this level is very rare. Retinal tumors have frequently systemic associations and in this case they are included in the phacomatoses syndrome; nevertheless, they can also occur as a nonsyndromic pathological disorder. Even if the incidence is rare and the variety limited, issuing a diagnosis proves difficult because retinal tumors look like other lesions or due to a low visibility of the ocular fundus. Hereunder we shall present you two cases of retinal tumors that were difficulty diagnosed and treated by surgical excision of the tumor. PMID- 15341099 TI - [Diagnostic and therapeutic difficulties in double depressors paralysis--DDP]. AB - Double depressor palsy is a rare monolateral condition showing a down gaze difficulty, associated sometimes with a superior lid retraction. MATERIAL AND METHOD: The paper presents the clinical situation and the surgical treatment in a patient with the symptoms of double depressors palsy. The surgical treatment consisted in deinsertion of the inferior oblique, recession of the medial rectus, and advancement of the lateral rectus 6.5 mm from the limbus. Surgery led to good alignment in the horizontal and vertical plan, but a slight limitation of abduction and of downward gaze persisted. PMID- 15341100 TI - [Unilateral amblypia with central fixation--occlusion treatment]. AB - Amblyopia refers to a decrease of vision, either unilaterally or bilaterally, for which no cause can be found by physical examination of visual pathway. Early detection and treatment is the the better way for this disease. There are many types of amblyopia treatment. Occlusion therapy has been the mainstay since 18th century. This treatment must be individualized and many clinical parameters will be follow up: rithm of occlusion, age of the child, different type of amblyopia, compliance of the child or parents, regular or irregular control. The success of the treatment will be when visual acuity is better with 2-3 lines, there is isoacuity or the alternance of unilateral esotropia, there is compliance from the child or parents. Treatment of amblyopia is difficult but has good results if parents, ophthalmologists or orthoptists work together. PMID- 15341101 TI - Clinical aspects of photodynamic therapy--Romanian experience. AB - Photodynamic therapy is an emerging method for local destruction of tissue by generating toxic oxygen species using light absorbed by an administered or an endogenously generated photosensitiser. It is a promising treatment for patients with cancer. More recently it has found increasing use as a method of therapy for non-cancerous illnesses. Following administration of a photosensitiser occurs an accumulation or retention in areas of cancer and disease relative to adjacent normal tissue. The photosensitiser is inactive until irradiated by light, following which cellular destruction occurs. This explanation is a good reason for the scientific and clinical interest in photodynamic therapy. PMID- 15341102 TI - [Paralytic lagophthalmos--treatment methods]. AB - Paralytic lagophthalmos has a negative effect on patient. A patient's appearance is grossly distorted and many ocular problems due to exposure and dryness may interfere with the patient's ability to perform an occupation. This paper describes the management of the paralytic lagophthalmos. The medical treatment adds lubricants and the surgical treatment provides a better closure for the eye. PMID- 15341103 TI - [Neuro-ophthalmologic aspects in antiphospholipid syndrome]. AB - Antiphospholipid syndrome is the one autoimmune noninflammatory syndrome affecting all blood vessels at any vascular segment, irrespective of the vascular size and topography. Defined in 1983 by Hughes, this syndrome represents the actual research theme in practically all medical fields, therefore the importance of antiphospholipid syndrome is recognized. The major aim of this article is to describe the neuro-ophthalmologic manifestations of the antiphospholipid syndrome, emphasizing the latest news about this syndrome and its impact on the retina and central nervous system. PMID- 15341104 TI - [The influence of cataract surgery on the efficacy of trabeculectomy in patients with open-angle glaucoma]. AB - PURPOSE: To study the influence of cataract surgery on the intraocular pressure decreasing effect of trabeculectomy in patients with open angle glaucoma. METHOD: Retrospective study that included two groups: --32 patients with open angle glaucoma, their intraocular pressure being controlled after filtering surgery; they were subject to cataract surgery at least 6 months after the trabeculectomy (the CT group)--44 patients with open angle glaucoma (primary or secondary) with intraocular pressure controlled after trabeculectomy (the T group). The evaluation included: preoperative intraocular pressure, early (at 2 months) and late (at 18 months) postoperative intraocular pressure, intra- and postoperative complications of the cataract surgery, the need for antiglaucoma medication. RESULTS: The cataract surgery in patients with trabeculectomy increases the intraocular pressure (initial mean IOP = 15.31 mmHg, final mean IOP = 18.53 mmHg, mean IOP = 3.22 mmHg). The intraocular pressure increase is more important after extracapsular extraction than after phacoemulsification (3.7 vs. 2.01 mmHg). Qualified therapeutical success was obtained in 81.25% of CT patients. In the T group the postoperative intraocular pressure maintained constant throughout the entire follow-up period. The number of additional antiglaucoma drugs was larger in the T group (0.7 versus 0.5). The patients that suffered intra and postoperative complications had increased intraocular pressure when compared to those without complications (mean IOP early = 3.1 mmHg, mean IOP late = 1.1 mmHg). CONCLUSIONS: Cataract surgery in patients with previous filtering surgery increases the intraocular pressure and the need for antiglaucoma drugs. The intraocular pressure rise was more important after extracapsular extraction than after phacoemulsification. Intra and postoperative complications result in a more important increase in intraocular pressure, in the first months after the cataract operation. PMID- 15341105 TI - [Phacoemulsification in white cataract]. AB - PURPOSE: The study presents the technical difficulties of the capsulorhexis and phacoemulsification in the white, intumescent cataract and the incidences occurred during the foldable IOL implantation with the Monarch II injector. METHODS: The clinical study comprises a number of 70 eyes with white intumescent cataract operated during 15 January-15 December 2003 by the same surgeon. The surgical intervention consisted of lens extraction by phacoemulsification with clear corneal incision, using the Monarch II injector for the foldable IOL implantation. In 56 eyes, the anterior capsule was stained with trypan blue. RESULTS: The intraoperative complications include: failed capsulorexis in 4 cases (5.71%), rupture of posterior capsule in 1 case (1.43%) and the conversion to classical extracapsular extraction in 3 cases (4.29%). IOL implantation was made with the help of the Monarch II injector in 67 cases (95.71%). The foldable IOL implantation in the bag was realised a single time in 44 cases (62.9%). We had the following incidences in the other cases: the upside down turning of the IOL during unfolding in 2 cases, the second haptic capture in the tunnel of the cartridge in 2 cases, the second haptic stucks outside or in the anterior chamber in 18 cases, the first haptic stays in the sulcus and the second one stays outside in 1 case. CONCLUSIONS: In eyes with 1 intumescent cataract we can perform the continuous curvilinear capsulorhexis using trypan blue, but also without staining the anterior capsule using mechanical or manual aspiration of the liquefied cortex and viscoelastic substances. The foldable intraocular lens implantation with the injector proves to be an easy and comfortable method after it has been practiced for a longer period of time if the right size of the cartridge is used. PMID- 15341106 TI - [Functional disturbances after acute angle closure glaucoma]. AB - This issue presents the results of a study on patients with acute angle closure glaucoma. We analyse the functional disturbances and their long term evolution, and we show some risk factors which lead to severe functional disturbances. PMID- 15341107 TI - [Presumed ocular histoplasmosis syndrome]. AB - Presumed ocular histoplasmosis syndrome is a macular hemorrhagic choroiditis of the young adult. The etiology of this syndrome is uncertain, but can be correlated with some positive reactions to histoplasmosis. The study was carried out on 12 patients, each of them was investigated by functional and objective ocular examinations, laboratory tests, fluorescein angiography and ultrasound examinations. The clinical signs of this syndrome of presumed ocular histoplasmosis were: macular focal lesions, subretinian hemorrhages, detachment of the retinal neurosensorial layer, presence of disseminated choroiditis scars and subretinian neovascularization. The correct evaluation of the choriocapillary perfusion is helpful to institute a proper therapy. PMID- 15341108 TI - [Primary juvenile glaucoma--clinical and genetic aspects]. AB - PURPOSE: We have done a retrospective study of juvenile glaucoma cases hospitalized between 1993-2002. MATERIAL AND METHOD: 52 primary juvenile glaucoma patients were available for our clinical and genealogical investigations. RESULTS: In this study we have analyzed the distribution of cases per year, the age of appearance of the diseases, and the distribution in function of the sex and the genealogical factors. Genealogic analyze was performed by statistical methods: Weinberg and penetrance. CONCLUSIONS: Under 16 years, juvenile glaucoma diagnosis was mostly established by examining the children for visual acuity disorders caused by different ametropies, especially progressive rapid myopia. Genealogical exams confirmed an autosomal recessive transmission with incomplete variable penetrance but the possibility of polygenic multifactorial heredity may be also considered. PMID- 15341109 TI - [Experimental animal studies in uveitis]. AB - The aim of this study is to observe that if inject the Streptococcus beta hemolytic group A in anterior chamber at rabbit bring about an acute uveitis. PMID- 15341110 TI - Nurses in managed care roles. AB - TNA and the Texas Department of Insurance collaborated on an integrated, education program that has provided benefits for all--graduate nursing students get an opportunity to learn about managed care issues and policies; TDI has gained the clinical expertise of the nursing profession in order to best evaluate quality of care. PMID- 15341111 TI - Rules proposal intended to clarify nurse staffing. PMID- 15341112 TI - Collaboration means win-win for nursing. PMID- 15341113 TI - Staff nurse participation is key. PMID- 15341115 TI - TNA takes zero tolerance position on physician abuse of RNs. PMID- 15341116 TI - Getting them in the door: faculty influence on students choosing nursing. PMID- 15341117 TI - More nursing students enrolled in school. PMID- 15341118 TI - OSHA sets final ergonomics rule. AB - After years of advocating for an ergonomic regulation through the Occupational Safety and Health Administration (OSHA), at last on November 14, 2000, the final standards were issued. The OSHA Ergonomic Standards are designed to prevent musculoskeletal injury by matching workplace conditions to job demands. This rule will provide long awaited relief to nurses in direct care who have worked for years without benefit, in most cases, of technology that could have prevented ergonomic injury. PMID- 15341119 TI - Playing in the big league at the Capitol. A nurse's view. PMID- 15341121 TI - Maintaining readiness, always. PMID- 15341122 TI - [Larval cells of sponge Halisarca dujardini (Demospongiae, Halisarcida). I. Separation of the cells and their aggregation properties]. AB - The majority of Demospongia members have a parenchymula larva, whose inner cells are similar to definite sponge cells. There are some contradictory opinions about the structure of larva in the marine sponge Halisarca dujardini: some authors deny the presence of inner cells, while other investigators consider this larva as a real parenchymula. We defined the larval cell types by cell separation in the percoll gradient and analysed their morphology and behaviour. The results showed the existence of 6 cell types in the parenchymula larva of H. dujardini, including 2 types of external flagellated cells. Each cell category differs in its morphology and aggregation ability. PMID- 15341123 TI - [Larval cells of sponge Halisarca dujardini (Demospongiae, Halisarcida). II. Some cell types marked by polyclonal antibodies]. AB - The recent morphological and experimental data concerning the involvement of flagellated cells in sponge larvae are contradictory and testify to or against the germinal layers inversion. A study of morphogenetic processes in sponges, in particular larval metamorphosis, is complicated by difficulties in identification and succession of certain cell types. It is possible to trace the destiny of flagellated and other larval cells by marking them with antibodies (AB) specified for each cell type. We separated larval and adult sponge cells of Halisarca dujardini in percoll density gradient and obtained polyclonal AB for the majority of these cell types. The protein pattern of larval flagellated cells differed significantly from that of other cell types. The major proteins of flagellated, collencyte-like and spherulous cells were used to raise the corresponding AB. Immunoblot showed all AB to be specific for certain proteins and suitable for immunofluorescence. The AB for flagellated cells reacted with the apical cytoplasm, but not with the flagellum, the AB for major protein of collencyte like cells stained cytoplasm granules. The AB for spherulous cells of the adult sponge reacted with larval spherulous cells supposed to be of maternal origin. So, the method of cell marking with specific polyclonal AB can facilitate analysis of the layers inversion problem, as well as elucidate the degree of cell differentiation in larvae, their conformity to cells of the adult sponge or their provisional destiny. PMID- 15341124 TI - [Increase in the number of contacts of endoplasmic reticulum with mitochondria and plasma membrane in yeast cells stimulated to division with He-Ne laser light]. AB - We studied the ultrastructure of Torulopsis sphaerica yeast cells irradiated with He-Ne laser (lambda = 632.8 nm, dose--460 J/m2) and then cultured for 6 h in the nutrient with 1% glucose by aeration. The length of membranes of endoplasmic reticulum (ER) and the number of its associations with mitochondria (M) and plasma membrane (PM) were measured on ultrathin sections. A distance of less than 50 nm between heterogeneous membranes was considered as an "association". The cells from irradiated cultures are characterized by the following features: 1) the length of cortical ER membranes in relation to cellular perimeter, and the length of perinuclear ER membranes in relation to nuclear perimeter increase, resp., by 21 and 79%; 2) the number of ER-PM associations per cellular section, and that per unit of PM length increase, resp., by 26 and 41%; 3) the number of ER-M association in relation to the total mitochondrial perimeter, and to perimitochondrial ER increase by 80 and 87%, resp. The latter may be associated with Ca2+ uptake by mitochondria associated with ER, which results in activation of respiration and ATP production. PMID- 15341125 TI - [Differentiation of the colon endocrine cells in mucosal epithelium of humans and some vertebrate animals]. AB - By electron microscopy, the mucosal epithelium of a distal part of the intestine of humans and other vertebrates (Rana temporaria, Lacerta vivipara, Columba livia, rabbit, rat, pig, cow) was examined. On the basis of the variety of cellular structure and the level of specialization of the involved elements a common regularity of ways of differentiation of endocrine cells was revealed. The updating of endocrine cells basically occurs as a result of differentiation of cambial cells through intermediate pre-enteroendocrine forms. Reorganization of exocrine epithelial cells is accomplished through "mixed" exo-endocrine cells. At the lower vertebrate animals the "mixed" cells are met much more frequently than in mammals, including man. In humans the greatest quantity of these cells is observed in the intra-uterine period of development and in cases of pathology. Ways of cytogenesis in endocrine cells are differently manfested in the course of ontogenesis and under broken ability to live. PMID- 15341126 TI - [Ultrastructure of root cup meristematic cells of Cucumis sativus L. seedlings under inhibition of root growth by synthetic cyclopentene beta,beta'-triketone]. AB - A study was made of the ultrastructure of root cap meristematic cells in the seedlings of Cucumis sativus L., whose growth was inhibited by the action of 2 acetyl-4-metylcyclopent-4-en-1,3-dione (AMCD). It is shown that the treatment of seeds by this substance at a concentration of 100 microg/ml leads to the main root inhibition by 73% and, as compared to the control, causes the highest changes in mitochondria that do not reach normal dimensions and undergo degeneration. AMCD caused significant changes in the cell walls that in test samples unusually became electron-dense. Deposits of osmiophylic substance, presumably consisting of terpenoid-like compounds, were often found in the cytoplasm of treated cells as well as in the extracellular space. In many cells, plasmalemma and amiloplast membranes underwent destructive changes. We discuss a possible relation between inhibitory effect of AMCD on the root growth, and its influence on the ultrastructure in investigated cells. PMID- 15341127 TI - [Investigation of daily mitotic activity of the birch, Betula pendula Roth]. AB - A study was made of the daily mitotic activity in the seedling root meristem of birch trees growing in an ecologically clear area--the biological station of Voronezh State University "Venevitinovo". The peak of mitotic activity was exposed at 9 a. m. (according to winter time). The rise of mitotic index was noted at 9 and 12 p. m. due to an increase in the share of cells being in the prophase stage, and to a high number of dividing cells with persistent nucleoli. A possibility of prolongation of the mitotic cycle time is supposed to be due to cell delay in prophase stage, which may be associated with anthropogenic and nature-climatic influences on the original trees themselves and on their seed progeny. This makes it possible to consider the investigated region as only conventionally clear, because of the availability of a high recreative pressure upon the trees. PMID- 15341128 TI - [Selective effect of epidermal growth factor on the endoribonuclease activity of different subpopulations of proteasomes from A431 cell line. Specificity of extracellular proteasome population]. AB - For the first time, it has been shown that population of proteasomes is heterogeneous in their RNAse activity. EGF exerts selective effect on different subpopulations of proteasomes. The RNAse activity of cytoplasmic proteasomes is induced under the influence of EGF on epidermoid carcinoma cell line A431. However, the activity of proteasomes isolated from culture medium and of nuclear proteasomes is inhibited by EGF. The above enzymatic activity has been shown to be specifically and selectively dependent on phosphorylation of proteasomal subunits in different subpopulations of proteasomes. Proteasome involvement in the coordinated control of specific messenger RNA molecules stability is suggested, and one of the mechanisms of this control might be an export of specific subpopulation of proteasomes from the cell. PMID- 15341129 TI - [The dynamics of microtubule repolymerization in a cell: rapid growth from the centrosome and slow recovery of free microtubules]. AB - According to the current view, the microtubule system in animal cells consists of two components: microtubules attached to the centrosome (these microtubules stretch radially towards the cell margin), and free microtubules randomly distributed in the cytoplasm without visible association with any microtubule organizing centers. The ratio of the two sets of microtubules in the whole microtubule array is under discussion. Addressing this question, we have analysed the recovery of microtubules in cultured Vero nucleated cells and cytoplasts, with and without centrosomes in these. Cells were fixed at different time points, and individual microtubules were traced on serial optical sections. During a slow recovery after cold treatment (4 degrees C, for 4 h; recovery at 30 degrees C) polymerization of microtubules started mainly from the centrosome. At early stages of recovery the share of free microtubules made about 10% of all microtubules, and their total length increased slower than the lenght of centrosome-attached microtubules. During a rapid recovery after nocodazole treatment (10 microg/ml, 2 h; recovery in drug-free medium at 37 degrees C), the share of free microtubules was about 35%, but their total length increased slower than the length of centrosome-attached microtubules. In 6-8 min (rapid recovery) or 12-16 min (slow recovery), tips of centrosomal microtubules reached the cell margin, and their increased density made it impossible to recognize individual microtubules. However, under the same conditions in cytoplasts without centrosomes the normal number of microtubules recovered only in 60 min, which enabled us to suppose that the complete recovery of microtubule system in the whole cells may be also rather long. When the first centrosomal microtubules reached the cell margin, the optical density of microtubules started to decrease from the centrosome region towards the cell margin, according to the exponential curve. Later on, the optical density in the centrosome region and near the cell margin remained at the same level, but microtubule density increased in the middle part of the cell, and in 45-60 min the plot of the optical density vs the distance from the centrosome became linear, as in control cells. Since no significant curling of microtubules occurs near the cell margin, the density of microtubules in the endoplasm may increase due only to polymerization of free microtubules. We suppose that in cultured cells the microtubule network recovery proceeds in two stages. At the initial stage, a rapid growth of centrosomal microtubules takes place in addition to the turnover of free microtubules with unstable minus ends. At the second stage, when microtubule growth from the centrosome becomes limited by the cell margin, a gradual extension of free microtubules occurs in the internal cytoplasm. PMID- 15341130 TI - [Prolactin-binding activity of bovine granulosa cells on luteinization and exposure to somatotropin in vitro]. AB - The influence of luteinization and bovine somatotropin (ST, 5-50 ng/ml) during cultivation of bovine granulosa cells on their ability to bind [125I]-labeled bovine prolactin (PRL) was studied. On the second day of cultivation in serumfree medium, granulosa cells from immature antral follicles underwent spontaneous luteinization, in both the absence and presence of ST. The level of [125I]-PRL specific binding to cells increased after two days of cultivation, with a negative correlation being revealed between estradiol production by the cells and their PRL-binding activity. At the same time, the addition of ST to the culture medium had no effect on the level of [125I]-PRL specific binding to native and luteinizing granulosa cells. The findings suggest a stimulatory influence of the luteal differentiation process on the PRL-binding activity of bovine granulosa cells, this influence is independent of the action of ST. PMID- 15341131 TI - [The role of Hsp70 chaperone in the reaction of human leukemic cells to anticancer drugs]. AB - Most of anti-tumor factors are designed to kill selectively cancer cells; in most cases this action is related to the ability of the above substances to induce apoptosis. One of potent anti-apoptotic mechanisms is based on Hsp70 protein. Since the level of this protein is often higher in malignant tumors than in normal tissues, the aim of this study was to establish whether the elevated Hsp70 content may influence the process of apoptosis induced by anti-tumor drugs in cancer cells population. The increase of intracellular content of Hsp70 in human leukemia U-937 cells was attained by a mild heat stress or by transfection of cells with the human hsp70 gene. The elevation of Hsp70 quantity, irrespective of the way it was performed, leads to the inhibition of apoptosis in cells treated with two substances, etoposide or adriamycin. The inhibition of apoptosis was accompanied with the reducing of the share of cells with fragmented nuclei and with the delay in caspase activation. It is suggested that in addition to the previously discovered targets, whose activity is suppressed by the Hsp70 chaperone, this protein can inhibit the activity of caspase-3 and -7; this delays the onset of apoptosis in part of a cancer cell population. PMID- 15341132 TI - [The influence of guanine nucleotides and protein kinase C on Ca2+ from intracellular stores of pigs oocytes stimulated by theophylline and cyclic AMP]. AB - Effect of guanine nucleotides and protein kinase C on Ca2+ exit from intracellular stores of pig oocytes, stimulated by theophylline and dbcAMP, was investigated using fluorescent dye chlortetracycline. Effect of cAMP on Ca2+ exit from intracellular stores of pig oocytes was not associated with activation of protein kinase C. In calcium-free medium, cAMP does not stimulate Ca2+ exit from intracellular stores of pig oocytes treated with GDP. In the presence of GDP, inhibition of protein kinase C activates Ca2+ exit from intracellular stores of pig oocytes on the action of cAMP. These data suggest the existence of different effects of guanine nucleotides on Ca2+ exit from intracellular stores of pig oocytes stimulated by cAMP. PMID- 15341133 TI - [Chromosomal aberrations and parameters of oxidative stress in Chernobyl clean-up workers in a remote period after ionizing radiation]. AB - This investigation was performed to test a supposition about the influence of disturbances in antioxidative system on cytogenetical parameters in Chernobyl' clean-up workers. It was shown that some cytogenetical parameters straightly correlated with the parameters of oxidative stress, while other cytogenetical parameters show just reverse correlations. Apparently, these relationships are not of cause-and-effect type, but they reflect a complex processes occurring in humans for a long period after radiation exposure. PMID- 15341134 TI - [Modulation of proliferation of peripheral blood lymphocytes after irradiation of volunteers with polychromatic visible and infrared light]. AB - Effects on human immune system of visible and infrared (IR) radiation, the dominating types of solar light on Earth, still remain poorly studied. In the present work, a small area of the volunteers' body surface was irradiated with polychromatic visible + IR polarized (VIP) light, whose spectral range is close to the natural one (400-3400 nm, 12 J/cm2), and spontaneous and phytohemagglutinin (PHA)-induced DNA syntheses were studied by radiometric method in lymphocytes (Lym) of peripheral blood. This Irradiation stimulated both spontaneous and PHA-induced DNA synthesis in Lym but only in volunteers with initially decreased parameters of synthesis (on average, by 2.5 and 2.7 times, respectively), which was recorded 24 h after irradiation of volunteers, and after a 72 h cultivation of separated mononuclears. In the parallel experiments, blood of each volunteer was irradiated in vitro. Besides, by modeling situation in vivo, when a small amount of transcutaneously photomodified blood contacts its much larger circulating volume, the irradiated and non-irradiated samples of autologous blood were mixed at a 1:10 volume ratio. In Lym with the initially decreased synthesis level, the spontaneous synthesis elevated by 2 and 3 times, respectively, whereas stimulation of PHA-synthesis was observed only after addition of the irradiated blood to the intact one (by 2.2 and 1.6 times, respectively). A high degree of positive correlation in changing the studied parameters is revealed in irradiation of blood in vivo and in vitro. This makes it possible to associate the light-stimulating effect on Lym of the entire circulating blood with transcutaneous photomodification of its small amounts, and with action of such blood on the rest of blood. A similarity in the direction and additivity of mitogenic effects of VIP light and PHA was revealed. The obtained data enable us to suggest that therapy employing polychromatic visible and IR light would promote presumably an increase in the number of Lym in peripheral blood and an enhancement of their response to antigenic stimulus. PMID- 15341135 TI - Changing dentate status of adults, use of dental health services, and achievement of national dental health goals in Denmark by the year 2000. AB - OBJECTIVES: This study analyzes the current profile of dentate status and use of dental health services among adults in Denmark at the turn of the millennium, assesses the impact on dentate status of sociodemographic factors and use of dental health services in adulthood and in childhood, and highlights the changes over time in dental health conditions among adults. Finally, the intention of the study was to evaluate the Danish dental health care system's level of achievement of the official goals for the year 2000 as formulated by the World Health Organization and the National Board of Health. The subjects of this study included a national representative sample of 16,690 Danish citizens aged 16 years and older (response rate=74.2%). A subsample (n=3,818) took part in a survey of dental care habits in childhood and prevalence of removable dentures; 66 percent of persons selected responded. METHODS: Personal interviews were used to collect information on dentate status, use of dental health services and living conditions; data on dental care habits in childhood and prevalence of removable dentures were collected by self-administered questionnaires. RESULTS: In all, 8 percent of interviewed persons were edentulous, while 80 percent had 20 or more natural teeth. At age 65-74 years, 27 percent were edentulous and 40 percent had 20 teeth or more; 58 percent wore removable dentures. Dentate status and prevalence of dentures were highly related to educational background and income, particularly for older age groups. Among persons interviewed, 80 percent paid regular dental visits and visits were most frequent among persons of high education and income. At age 35-44 years 95 percent had participated in regular dental care in childhood compared to 49 percent of 65-74-year-olds. Multivariate analyses revealed that sociobehavioral factors had significant effects on dentate status. CONCLUSIONS: Compared to similar studies carried out in 1987 and 1994, the present survey indicates a positive trend of improved dentate status in adult Danes in general and regular use of dental health services increased considerably over time. The WHO goals for better dental health by the year 2000 were achieved for 35-44-year-olds, whereas the goal of more people with functional dentitions at age 65 years or older was not achieved. It remains a challenge to the Danish dental health system to help even out the social inequalities in dental health. PMID- 15341136 TI - Dental caries status and need for dental treatment of Pennsylvania public school children in grades 1, 3, 9, and 11. AB - OBJECTIVES: This cross-sectional study was designed to determine the caries status and provide a general evaluation of the level of dental treatment need of Pennsylvania public school children in grades 1, 3, 9, and 11 on a statewide and regional basis. METHODS: Between September 1998 and May 2000, caries status and treatment need were assessed using a school-based dental examination, performed on a representative sample (n=6,040) of public school children in grades 1, 3, 9, and 11 (age range=6 to 21 years). Children's caries status in the primary and permanent dentition was assessed. Need for treatment was scored on a three-level categorical scale--no treatment need identified, routine treatment need, and urgent treatment need--and was based on the presence and severity of caries and other oral conditions. Population estimates of the prevalence of untreated dental caries, DMFT and dft scores, and treatment need were calculated by grade and geographically, using the six Pennsylvania health districts and the cities of Pittsburgh and Philadelphia. The inequality of caries distribution in the population was assessed for both permanent and primary caries using Lorenz curves and Gini coefficients. RESULTS: Dental caries has remained highly prevalent among Pennsylvania's public school children. Caries levels varied considerably by health districts and city. Urgent treatment needs were significant and also varied by health district and city. CONCLUSIONS: Dental caries remains the most prevalent disease affecting Pennsylvania's schoolchildren. Caries status varies significantly by region of the state, suggesting that environmental, social, and demographic contextual factors may be important determinants of disease prevalence. PMID- 15341137 TI - Life-style correlates of tooth loss in an adult Midwestern population. AB - OBJECTIVES: To describe life-style correlates of tooth loss in a representative rural American population. METHODS: Information on tooth loss as well as past medical history and life-style factors was obtained in a well-defined cohort of 2,764 persons 53-96 years of age in Beaver Dam, WI (1998-2000). RESULTS: There were 1,992 (68.2%) persons missing some and 447 (15.3%) missing all of their teeth. In univariable analyses, age, cigarette smoking, heavy drinking, education, multivitamin use, and diabetes status were associated with tooth loss. Tooth loss was associated with poorer self-rated health and with difficulty eating solid food and inability to enjoy some food. In multivariable models age, education, smoking, heavy drinking, and diabetes were significantly associated with tooth loss. CONCLUSIONS: Tooth loss is common in older persons and is associated with many risk factors including education, smoking, and heavy drinking. It is possible that modifying these may influence risk of tooth loss. PMID- 15341138 TI - Prevalence and handedness correlates of recurrent aphthous stomatitis in the Turkish population. AB - OBJECTIVES: The present study was designed to determine the relationship between recurrent aphthous stomatitis (RAS) and handedness, and to assess the prevalence of RAS in the Turkish population. METHODS: The present study was conducted among 11,360 persons (5,705 males and 5,655 females) with a mean age of 30.4 years. A questionnaire focusing on handedness was administered to these patients. Handedness was assessed according to the Edinburgh Handedness Inventory. The diagnosis of RAS was made on the basis of clinical appearance, location, and the patient's health history. RAS information of patients was collected by means of a data form specifically designed for this study. Subjects were informed about RAS. We took anamneses and filled out the investigation forms. Apart from patients with registration of current aphthae (average point prevalence, APP), patients who had a past two-year history (self-reported two-year prevalence, SRTP) of the lesion were also included in this study. Data were analyzed using the chi-square and logistic regression tests. RESULTS: The prevalence of RAS (APP) was 2.7 percent and that of a history of RAS (SRTP) 22.8 percent. Thus, the total prevalence (APP+SRTP), including present lesions and a two-year history (SRTP), was 25.5 percent. Adjusted results showed that females, left-handers, 10-30-year olds, and nonsmokers were 1.53, 1.69, 2.05, and 1.61 times more likely to have RAS (APP+SRTP) than males, right-handers, 31-50-year-olds, and smokers, respectively (P<.0001). CONCLUSION: The present study suggests that left handedness appears to be a predictor factor for RAS. PMID- 15341139 TI - Association between mandibular retrognathia and TMJ disorders in adult females. AB - OBJECTIVE: This study assesses the association between temporomandibular joint (TMJ) disorders and mandibular retrognathia. METHODS: We conducted a case-control study among women, aged 18-70 years, recruited from Kaiser Permanente Northwest Division (KPNW, Portland, OR) and Group Health Cooperative of Puget Sound (GHC, Seattle, WA). Cases (N= 160) were women seeking treatment for TMJ disorders at KPNW and GHC clinics. Controls (N=151) were women without TMJ disorders, selected from among adult female dental hygiene patients at KPNW or GHC enrollees. Case status was determined by questionnaire; mandibular sagittal position (orthognathic, mildly retrognathic, severely retrognathic, or prognathic) was measured using digitized facial photographs. Odds ratios (OR) and associated 95 percent confidence intervals (CI) estimated the magnitude of the association between TMJ disorders and mandibular sagittal position. RESULTS: Women with TMJ disorders were 4.0 times (95% CI=1.5, 10.8) more likely than controls to have severe retrognathia; no association with mild retrognathia or prognathia was seen. Results were similar when cases were restricted to those with recent onset of TMJ pain (OR=6.3; 95% CI=1.8, 21.8). CONCLUSIONS: We found a strong association between TMJ disorders and severe mandibular retrognathia in adult females. In some women this likely resulted from TMJ disorders influencing mandibular development over time. However, among a subset of women, our data support the reverse hypothesis--that severe mandibular retrognathia may influence the development of TMJ disorders. Despite this strong association, abnormal mandibular position contributed only a small portion to the overall rate at which women sought treatment for TMJ disorders. PMID- 15341140 TI - Efficacy of educational interventions targeting primary care providers' practice behaviors: an overview of published systematic reviews. AB - OBJECTIVES: Primary care providers (e.g., family physicians, pediatricians, registered nurses, physician assistants, and nurse practitioners) could play a pivotal role in the provision of preventive services, especially for very young children (younger than 3 years old) and population groups with limited access to dental care. Given the current problems with access to dental care among low income Americans, we contend there is a need to involve nondental primary health care providers in screening for and preventing oral health problems. The objective of this overview is to present findings from systematic reviews on the efficacy of continuing medical education, printed educational material, academic outreach, reminders, and local opinion leaders on the adoption of new knowledge and practices by primary care providers. METHODS: A search was conducted using the Cochrane Library and MEDLINE. The search aimed to locate systematic reviews published between January 1988 and March 2003. Two researchers independently extracted data and assessed study quality using a modified version of the QUOROM statement. RESULTS: Eleven systematic reviews were included in this overview. The evidence from the included systematic reviews showed that formal continuing medical education (CME) and distributing educational materials did not effectively change primary care providers' behaviors. There are effective interventions available to increase knowledge and change behaviors of primary care providers, such as small group discussion, interactive workshops, educational outreach visits, and reminders. CONCLUSION: There is a limited knowledge base on the efficacy of the selected interventions on oral health screening by primary care providers. Considering the potential role of primary care providers in improving oral health of underserved populations, this research area should receive more attention. PMID- 15341141 TI - Oral cancer knowledge and examination experiences among North Carolina adults. AB - OBJECTIVE: This study assesses knowledge of oral cancer risk factors, clinical signs, and oral cancer examination experience among North Carolina adults. METHODS: A statewide random digit dial, computer-assisted telephone interview was conducted in 2002. Data from 1,096 respondents, with a response rate of 62 percent, were poststratified to 2000 census data by sex, race, and age group to produce population-based estimates. Knowledge of one sign of oral cancer, four or more risk factors for oral cancer, and having ever had an oral cancer examination were compared in logistic regression models using normalized weights. RESULTS: Fourteen (95% confidence interval [CI] +/-2) percent of adults had never heard of oral or mouth cancer. Risk factor knowledge was high for 56 percent (95% CI+/-3) and associated in a logistic regression model with younger age, feeling personal factors cause cancer, and nonuse of snuff. One sign of oral cancer (sore/lesion, red or white patch in mouth, and bleeding in the mouth) was correctly identified by 53 percent (95% CI+/-3) with significantly more correct responses from younger people, nonsmokers, and some college education. Only 29 percent (95% CI+/-3) reported ever having had an oral cancer examination when this procedure was described. Most respondents reported exams performed by dentists. In a weighted logistic regression model, older age, being dentate, nonsmokers, alcohol users, and those with some college education were significantly more likely to report having ever had an oral cancer examination. CONCLUSIONS: Although there is moderate knowledge of signs and risk factors for oral cancer among North Carolina adults, knowledge deficits remain. Oral cancer examinations need to be increased, particularly among tobacco smokers. PMID- 15341142 TI - Association of denture use with sleep-disordered breathing among older adults. AB - OBJECTIVE: This study investigates the relationship between sleep-disordered breathing and denture use. METHODS: This was a cross-sectional study of community dwelling older adults. Information about denture use was obtained using a questionnaire. Ambulatory sleep recording in subjects' homes was performed using Embletta PDS (Medcare, Iceland). Chi-square tests and logistic regression analysis were used for statistical analysis. RESULTS: A total of 58 subjects completed the study. The mean apnea hypopnea (AHI) index was 15.1+/-16.1. Twenty two subjects (38%) used dentures and most removed them before sleep. There was significant association between denture use and AHI > or = 15 per hour of sleep (odds ratio [OR]=6.29; 95% confidence interval [CI]= 1.71, 23.22; P=.006). CONCLUSIONS: This preliminary study found an association between sleep-disordered breathing and denture use, which may represent a proxy for a relationship between sleep-disordered breathing and edentulism. Given the common occurrence of both conditions among older adults, the observed relationship warrants a more detailed investigation of the mechanisms whereby loss of teeth leads to upper airway closure during sleep. PMID- 15341143 TI - The association between environmental tobacco smoke and primary tooth caries. AB - OBJECTIVE: Environmental tobacco smoke (ETS) has been associated with a number of negative health outcomes for exposed children. The goal of this study was to assess the association between ETS and dental caries in a pediatric population. METHODS: This study included 637 Iowa Fluoride Study children whose parents provided socioeconomic information, completed at least three questionnaires during the first year of life, and had a primary dentition exam at age 4-7 years. Households reporting in all questionnaires that someone smoked in the home were categorized as regularly smoking homes. Socioeconomic status (SES) was divided into three groups (low, middle, and high) based on family income and mother's education. Children were classified as having caries if any of the primary teeth had fillings or cavitated lesions at the primary dentition exam. RESULTS: Overall, children residing in regularly smoking homes had a higher prevalence of caries. For the middle SES group and overall, the children from smoking homes had a significantly higher prevalence of caries compared to nonregular/nonsmoking homes (52% vs 24%, P=.05 and 44% vs 25%, P=.002, respectively). After adjusting for age, SES, toothbrushing frequency, total ingested fluoride, and combined intake of soda pop and powdered drink beverages, the relationship of smoking and caries still remained significant (odds ratio [OR]=3.38; P=.001). CONCLUSIONS: Environmental tobacco smoke was associated with an increased risk of caries among children. PMID- 15341144 TI - Adaptive evolution of asexual populations under Muller's ratchet. AB - We study the population genetics of adaptation in nonequilibrium haploid asexual populations. We find that the accumulation of deleterious mutations, due to the operation of Muller's ratchet, can considerably reduce the rate of fixation of advantageous alleles. Such reduction can be approximated reasonably well by a reduction in the effective population size. In the absence of Muller's ratchet, a beneficial mutation can only become fixed if it creates the best possible genotype; if Muller's ratchet operates, however, mutations initially arising in a nonoptimal genotype can also become fixed in the population, since the loss of the least-loaded class implies that an initially nonoptimal background can become optimal. We show that, while the rate at which adaptive mutations become fixed is reduced, the rate of fixation of deleterious mutations due to the ratchet is not changed by the presence of beneficial mutations as long as the rate of their occurrence is low and the deleterious effects of mutations (s(d)) are higher than the beneficial effects (s(a)). When s(a) > s(d), the advantage of a beneficial mutation can outweigh the deleterious effects of associated mutations. Under these conditions, a beneficial allele can drag to fixation deleterious mutations initially associated with it at a higher rate than in the absence of advantageous alleles. We propose analytical approximations for the rates of accumulation of deleterious and beneficial mutations. Furthermore, when allowing for the possible occurrence of interference between beneficial alleles, we find that the presence of deleterious mutations of either very weak or very strong effect can marginally increase the rate of accumulation of beneficial mutations over that observed in the absence of such deleterious mutations. PMID- 15341145 TI - Sexual conflict and protein polymorphism. AB - Sexual conflict, where male and female reproductive interests differ, is probably widespread and often mediated by male or sperm proteins and female or egg proteins that bind to each other during mating or fertilization. One potential consequence is maintenance of polymorphism in these proteins, which might result in reproductive isolation between sympatric subpopulations. I investigate the conditions for polymorphism maintenance in a series of mathematical models of sexual conflict over mating or fertilization frequency. The models represent a male or sperm ligand and a female or egg receptor, and they differ in whether expression of either protein is haploid or diploid. For diploid expression, the conditions imply that patterns of dominance, which involve neither overdominance nor underdominance, can determine whether polymorphism is maintained. For example, suppose ligand expression is diploid, and consider ligand alleles L1 and L2 in interactions with a given receptor genotype; if L1/L1 males are fitter than L2/L2 males in these interactions, then polymorphism is more likely to be maintained when L1/L2 males more closely resemble L1/L1 males in these interactions. Such fitter-allele dominance might be typical of a ligand or its receptor due to their biochemistry, in which case polymorphism might be typical of the pair. PMID- 15341146 TI - Quantifying patterns in the evolution of reproductive isolation. AB - We present a likelihood-based statistical method for examining the pattern or rate of evolution of reproductive isolation. The method uses large empirical datasets to estimate, for a given clade, the average duration of two phases in the divergence of populations. The first phase is a lag phase and refers to the period during which lineages diverge but no detectable reproductive isolation evolves. The second is an accumulation phase, referring to the period during which the magnitude of reproductive isolation between diverging lineages increases. The pattern of evolution is inferred from the relative durations of these two phases. Results of analyses of postzygotic isolation data indicate significant differences among taxa in the pattern of evolution of postzygotic isolation that are consistent with predictions based on genetic differences among these groups. We also examine whether the evolution of postzygotic isolation is best explained by either of two models for the rate of accumulation: a linear model or a quadratic function as may be suggested by recent studies. Our analysis indicates that the appropriateness of either model varies among taxa. PMID- 15341147 TI - Experimental demonstration of a causal relationship between heterogeneity of selection and genetic differentiation in quantitative traits. AB - Comparisons of estimates of genetic differentiation at molecular markers (F(ST)) and at quantitative traits (Q(ST)) are a means of inferring the level and heterogeneity of selection in natural populations. However, such comparisons are questionable because they require that the influence of drift and selection on Q(ST) be detectable over possible background influences of environmental or nonadditive genetic effects on Q(ST)-values. Here we test this using an experimental evolution approach in metapopulations of Arabidopsis thaliana experiencing different levels of drift and selection heterogeneity. We estimated the intensity and heterogeneity of selection on morphological and phenological traits via selection differentials. We demonstrate that Q(ST)-values increased with increasing selection heterogeneity when genetic drift was limited. The effect of selection on Q(ST) was thus detectable despite significant genotype-by environment interactions that most probably biased the estimates of genetic differentiation. Although they cannot be used as a direct validation of the conclusions of prior studies, our results strongly support both the relevance of Q(ST) as an estimator of genetic differentiation and the role of local selection in shaping the genetic differentiation of natural populations. PMID- 15341148 TI - Evolutionary potential of Chamaecrista fasciculata in relation to climate change. I. Clinal patterns of selection along an environmental gradient in the great plains. AB - Climate change will alter natural selection on native plant populations. Little information is available to predict how selection will change in the future and how populations will respond. Insight can be obtained by comparing selection regimes in current environments to selection regimes in environments similar to those predicted for the future. To mimic predicted temporal change in climate, three natural populations of the annual legume Chamaecrista fasciculata were sampled from a climate gradient in the Great Plains and progeny of formal crosses were reciprocally planted back into common gardens across this climate gradient. In each garden, native populations produced significantly more seed than the other populations, providing strong evidence of local adaptation. Phenotypic selection analysis conducted by site showed that plants with slower reproductive development, more leaves, and thicker leaves were favored in the most southern garden. Evidence of clinal variation in selection regimes was also found; selection coefficients were ordered according to the latitude of the common gardens. The adaptive value of native traits was indicated by selection toward the mean of local populations. Repeated clinal patterns in linear and nonlinear selection coefficients among populations and within and between sites were found. To the extent that temporal change in climate into the future will parallel the differences in selection across this spatial gradient, this study suggests that selection regimes will be displaced northward and different trait values will be favored in natural populations. PMID- 15341149 TI - Evolutionary potential of Chamaecrista fasciculata in relation to climate change. II. Genetic architecture of three populations reciprocally planted along an environmental gradient in the great plains. AB - Climate change will test the evolutionary potential of populations. Information regarding the genetic architecture within and among populations is essential for prediction of evolutionary outcomes. However, little is known about the distribution of genetic variation for relevant traits in natural populations or alteration of genetic architecture in a changing environment. In this study, pedigreed families from three populations of the annual prairie legume Chamaecrista fasciculata were reciprocally transplanted in three environments across a broad latitudinal range in the Great Plains. The underlying premise of this work is that northern populations will in the future experience climates similar to current-day climates further south. Estimates of narrow-sense heritability ranged from 0.053 to 0.481, suggesting the potential for evolutionary change is possible for most traits. In general, the northern population harbored less genetic variation and had lower heritability for traits than the southern population. This population also experienced large reductions in fitness, as measured by estimated lifetime fecundity, when raised in either the intermediate or the southern climate, whereas the difference between the intermediate and southern population was less extreme. For fecundity, the pattern of cross-environment additive genetic correlations was antagonistic to evolutionary change in four of six cases when native and nonnative sites were compared. Six additional antagonistic positive correlations were found for the rate of phenological development and leaf thickness. Overall, the data suggest that if climate changes as predicted, the northern population will face a severe evolutionary challenge in the future because of low heritabilities, cross environment genetic correlations antagonistic to selection, and demographic instability due to lower seed production in a hotter and drier climate. PMID- 15341150 TI - Phylogeography of the planktonic chaetognath Sagitta setosa reveals isolation in European seas. AB - Numerous planktonic species have disjunct distribution patterns in the world's oceans. However, it is unclear whether these are truly unconnected by gene flow, or whether they are composed of morphologically cryptic species. The marine planktonic chaetognath Sagitta setosa Muller has a discontinuous geographic distribution over the continental shelf in the northeastern Atlantic, Mediterranean Sea, and Black Sea. Morphological variation between these populations has been described, but overlaps and is therefore unsuitable to determine the degree of isolation between populations. To test whether disjunct populations are also genetically disjunct, we sequenced a 504-bp fragment of mitochondrial DNA comprising the cytochrome oxidase II region of 86 individuals. Sequences were highly variable; each represented a different haplotype. Within S. setosa, sequence divergence ranged from 0.2 to 8.1% and strong phylogeographic structure was found, with four main groups corresponding to the northeastern Atlantic, Mediterranean Sea (including Ligurian Sea, Tyrrhenian Sea and Gulf of Gabes), Adriatic Sea, and Black Sea. Two of these (Atlantic and Black Sea) were resolved as monophyletic clades, thus gene flow between disjunct populations of S. setosa has been extremely limited and lineage sorting has taken place. The deepest divergence was between Atlantic and Mediterranean/Black Sea populations followed by a split between Mediterranean and Black Sea populations. The Mediterranean/Black Sea clade comprised three groups, with the Adriatic Sea as the most likely sister clade of the Black Sea. These data are consistent with a colonization of the Black Sea from the Mediterranean. Furthermore, a possible cryptic species was found in the Black Sea with 23.1% sequence divergence from S. setosa. Two possibilities for the evolutionary origin of this species are proposed, namely, that it represents a relict species from the ancient Paratethys, or that it represents another chaetognath species that colonized the Black Sea more recently. Even though the exact timing of disjunction of S. setosa populations remains unclear, on the basis of the geological and paleoclimatic history of the European basins and our estimates of net nucleotide divergence, we suggest that disjunct populations arose through vicariance resulting from the cyclical changes in temperature and sea levels during the Pleistocene. We conclude that these populations have remained disjunct, not because of limited dispersal ability, but because of the inability to maintain viable populations in suboptimal, geographically intermediate areas. PMID- 15341151 TI - Phylogenetic effects, the loss of complex characters, and the evolution of development in calyptraeid gastropods. AB - Despite considerable theoretical and empirical work on the population genetic effects of mode of development in benthic marine invertebrates, it is unclear what factors generate and maintain interspecific variation in mode of development and few studies have examined such variation in a phylogenetic context. Here I combine data on mode of development with a molecular phylogeny of 72 calyptraeid species to test the following hypotheses about the evolution of mode of development: (1) Is the loss of feeding larvae irreversible? (2) Is there a phylogenetic effect on the evolution of mode of development? (3) Do embryos of direct-developing species lose the structures necessary for larval feeding and swimming and, if so, is the degree of embryonic modification correlated with the genetic distance between species? The results of these analyses suggest that mode of development evolves rapidly and with little phylogenetic inertia. There are three cases of the possible regain of feeding larvae, in all cases from direct development with nurse eggs. It appears that species with planktotrophic, lecithotrophic, or direct development with nurse eggs all have equal evolutionary potential and retain the possibility of subsequent evolution of a different mode of development. However, species with direct development from large yolky eggs appear to be subject to phylogenetic constraints and may not be able to subsequently evolve a different mode of development. Finally, species that have more recently evolved direct development have less highly modified embryos than older direct-developing species. Since species with nurse eggs generally have fewer embryonic modifications than those from large yolky eggs, this embryological difference may be the underlying cause of the difference in evolutionary potential. PMID- 15341152 TI - Convergence to a novel environment: comparative method versus experimental evolution. AB - Laboratory adaptation allows researchers to contrast temporal studies of experimental evolution with comparative studies. The comparative method is here taken to mean the inference of microevolutionary processes from comparisons among contemporaneous populations of diverse origins, from one or multiple species. The data contrasted here come from Drosophila subobscura populations that were introduced to the laboratory at several different times and from two different locations. Two questions were addressed. First, can we correctly infer evolutionary dynamics from comparative data collected simultaneously from disparate populations? In most cases, we could, except for the character of starvation resistance. Second, are the evolutionary dynamics inferred from the comparative approach similar to those revealed by temporal studies of experimental evolution? For fecundity characters, they were. Overall the results show that both comparative and temporal studies are useful, though the former can be uninformative for characters with complex evolutionary trajectories. PMID- 15341153 TI - Parasites and sexual reproduction in psychid moths. AB - Persistence of sexual reproduction among coexisting asexual competitors has been a major paradox in evolutionary biology. The number of empirical studies is still very limited, as few systems with coexisting sexual and strictly asexual lineages have been found. We studied the ecological mechanisms behind the simultaneous coexistence of a sexually and an asexually reproducing closely related species of psychid moth in Central Finland between 1999 and 2001. The two species compete for the same resources and are often infected by the same hymenopteran parasitoids. They are extremely morphologically and behaviorally similar and can be separated only by their reproductive strategy (sexual vs. asexual) or by genetic markers. We compared the life-history traits of these species in two locations where they coexist to test predictions of the cost-of-sex hypothesis. We did not find any difference in female size, number of larvae, or offspring survival between the sexuals and asexuals, indicating that sexuals are subject to cost of sex. We also used genetic markers to check and exclude the possibility of Wolbachia bacteria infection inducing parthenogenesis. None of the samples was infected by Wolbachia and, thus, it is unlikely that these bacteria could affect our results. We sampled 38 locations to study the prevalence of parasitoids and the moths' reproductive strategy. We found a strong positive correlation between prevalence of sexual reproduction and prevalence of parasitoids. In locations where parasitoids are rare asexuals exist in high densities, whereas in locations with a high parasitoid load the sexual species was dominant. Spatial distribution alone does not explain the results. We suggest that the parasite hypothesis for sex may offer an explanation for the persistence of sexual moths in this system. PMID- 15341154 TI - Quantitative genetics of continuous reaction norms: thermal sensitivity of caterpillar growth rates. AB - A continuous reaction norm or performance curve represents a phenotypic trait of an individual or genotype in which the trait value may vary with some continuous environmental variable. We explore patterns of genetic variation in thermal performance curves of short-term caterpillar growth rate in a population of Pieris rapae. We compare multivariate methods, which treat performance at each test temperature as a distinct trait, with function-valued methods that treat a performance curve as a continuous function. Mean growth rate increased with increasing temperatures from 8 to 35 degrees C, was highest at 35 degrees C, and declined at 40 degrees C. There was substantial and significant variation among full-sib families in their thermal performance curves. Estimates of broad-sense genetic variances and covariances showed that genetic variance in growth rate increased more than 30-fold from low (8-11 degrees C) to high (35-40 degrees C) temperatures, even after differences in mean growth rate across temperatures were removed. Growth rate at 35 and 40 degrees C was negatively correlated genetically, suggesting a genetic trade-off in growth rate at these temperatures; this trade-off may represent either a generalist-specialist trade-off and/or variation in the optimal temperature for growth. The estimated genetic variance covariance function (G function), the function-valued analog of the variance covariance matrix (G matrix), was quite bumpy compared with the estimated G matrix; and results of principal component analyses of the G function were difficult to interpret. The use of orthogonal polynomials as the basis functions in current function-valued estimation methods may generate artifacts when the true G function has prominent local features, such as strong negative covariances at nearby temperatures (e.g. at 35 and 40 degrees C); this may be a particular issue for thermal performance curves and other highly nonlinear reaction norms. PMID- 15341155 TI - Negative maternal or paternal effects on tactic inheritance under a conditional strategy. AB - Alternative behavioral and life-history tactics are common in animal populations. The conditional strategy model provides a powerful explanation for the evolution and persistence of such tactics, as it allows alternative tactics to be perpetuated even if there is tactic inheritance and tactics yield unequal mean fitness. In many biological systems negative maternal or paternal effects complicate the inheritance of condition and, hence, the inheritance of alternative tactics. Indeed, the inheritance of condition may result in the alternation of tactics across generations. In this paper, we show that the conditional strategy is robust to these effects on progeny condition. There is a unique and stable proportion of tactics under standard inheritance and unequal tactic fitness, and these two important properties of the conditional strategy hold even if negative maternal or paternal effects on progeny condition cause tactics to alternate across generations. However, the dynamics of tactic proportions pursuant to a perturbation of the equilibrium tactic proportions depend on the form of tactic inheritance. An application of our theoretical results to data from a population of smallmouth bass (Micropterus dolomieu) in which negative paternal effects dictate progeny condition reveals that age at first reproduction in males alternates regularly across generations. Furthermore, the model indicates that the population would return rapidly to equilibrium if the proportions of males that mature early or late in life were perturbed from the equilibrium within the system. This example shows how the model of the conditional strategy can be used to gain insight into tactic dynamics in situations where some of the model parameters are difficult or impossible to measure empirically. PMID- 15341156 TI - When vicars meet: a narrow contact zone between morphologically cryptic phylogeographic lineages of the rainforest skink, Carlia rubrigularis. AB - Phylogeographic analyses of the fauna of the Australian wet tropics rainforest have provided strong evidence for long-term isolation of populations among allopatric refugia, yet typically there is no corresponding divergence in morphology. This system provides an opportunity to examine the consequences of geographic isolation, independent of morphological divergence, and thus to assess the broader significance of historical subdivisions revealed through mitochondrial DNA phylogeography. We have located and characterized a zone of secondary contact between two long isolated (mtDNA divergence > 15%) lineages of the skink Carlia rubrigularis using one mitochondrial and eight nuclear (two intron, six microsatellite) markers. This revealed a remarkably narrow (width < 3 km) hybrid zone with substantial linkage disequilibrium and strong deficits of heterozygotes at two of three nuclear loci with diagnostic alleles. Cline centers were coincident across loci. Using a novel form of likelihood analysis, we were unable to distinguish between sigmoidal and stepped cline shapes except at one nuclear locus for which the latter was inferred. Given estimated dispersal rates of 90-133 m x gen(-1/2) and assuming equilibrium, the observed cline widths suggest effective selection against heterozygotes of at least 22-49% and possibly as high as 70%. These observations reveal substantial postmating isolation, although the absence of consistent deviations from Hardy-Weinberg equilibrium at diagnostic loci suggests that there is little accompanying premating isolation. The tight geographic correspondence between transitions in mtDNA and those for nuclear genes and corresponding evidence for selection against hybrids indicates that these morphologically cryptic phylogroups could be considered as incipient species. Nonetheless, we caution against the use of mtDNA phylogeography as a sole criterion for defining species boundaries. PMID- 15341157 TI - Evolution of color variation in dragon lizards: quantitative tests of the role of crypsis and local adaptation. AB - Many animal species display striking color differences with respect to geographic location, sex, and body region. Traditional adaptive explanations for such complex patterns invoke an interaction between selection for conspicuous signals and natural selection for crypsis. Although there is now a substantial body of evidence supporting the role of sexual selection for signaling functions, quantitative studies of crypsis remain comparatively rare. Here, we combine objective measures of coloration with information on predator visual sensitivities to study the role of crypsis in the evolution of color variation in an Australian lizard species complex (Ctenophorus decresii). We apply a model that allows us to quantify crypsis in terms of the visual contrast of the lizards against their natural backgrounds, as perceived by potential avian predators. We then use these quantitative estimates of crypsis to answer the following questions. Are there significant differences in crypsis/conspicuousness among populations? Are there significant differences in crypsis conspicuousness between the sexes? Are body regions "exposed" to visual predators more cryptic than "hidden" body regions? Is there evidence for local adaptation with respect to crypsis against different substrates? In general, our results confirmed that there are real differences in crypsis/conspicuousness both between populations and between sexes; that exposed body regions were significantly more cryptic than hidden ones, particularly in females; and that females, but not males, are more cryptic against their own local background than against the background of other populations [corrected]. Body regions that varied most in contrast between the sexes and between populations were also most conspicuous and are emphasized by males during social and sexual signaling. However, results varied with respect to the aspect of coloration studied. Results based on chromatic contrast ("hue" of color) provided better support for the crypsis hypothesis than did results based on achromatic contrast ("brightness" of color). Taken together, these results support the view that crypsis plays a substantial role in the evolution of color variation and that color patterns represent a balance between the need for conspicuousness for signaling and the need for crypsis to avoid predation. PMID- 15341158 TI - Developmental success, stability, and plasticity in closely related parthenogenetic and sexual lizards (Heteronotia, Gekkonidae). AB - The developmental trajectory of an organism is influenced by the interaction between its genes and the environment in which it develops. For example, the phenotypic traits of a hatchling reptile can be influenced by the organism's genotype, by incubation temperature, and by genetically coded norms of reaction for thermally labile traits. The evolution of parthenogenesis provides a unique opportunity to explore such effects: a hybrid origin of this trait in vertebrates modifies important aspects of the genotype (e.g., heterozygosity, polyploidy) and may thus impact not only on the phenotype generally, but also on the ways in which incubation temperature affects expression of the phenotype. The scarcity of vertebrate parthenogenesis has been attributed to developmental disruptions, but previous work has rarely considered reaction norms of embryogenesis in this respect. We used closely related sexual and asexual races of the Australian gecko Heteronotia binoei, which include those with multiple origins of parthenogenesis, to explore the ways in which reproductive modes (sexual, asexual), incubation temperatures (24, 27, and 30 degrees C), and the interaction between these factors affected hatchling phenotypes. The hatchling traits we considered included incubation period, incidence of deformities, hatchling survivorship, body size and shape, scalation (including fluctuating asymmetry), locomotor performance, and growth rate. Developmental success was slightly reduced (higher proportion of abnormal offspring) in parthenogenetic lineages although there was no major difference in hatching success. Incubation temperature affected a suite of traits including incubation period, tail length, body mass relative to egg mass, labial scale counts, running speed, growth rate, and hatchling survival. Our data also reveal an interaction between reproductive modes and thermal regimes, with the phenotypic traits of parthenogenetic lizards less sensitive to incubation temperature than was the case for their sexual relatives. Thus, the evolution of asexual reproduction in this species complex has modified both mean hatchling viability and the norms of reaction linking hatchling phenotypes to incubation temperature. Discussions on the reasons why parthenogenetic organisms are scarce in nature should take into account interactive effects such as these; future work could usefully try to tease apart the roles of parthenogenesis, its hybrid origin (and thus effects on ploidy and heterozygosity, etc.), and clonal selection in generating these divergent embryonic responses. PMID- 15341159 TI - Barriers to sympatry between avian sibling species (Paridae: Baeolophus) in local secondary contact. AB - Range limits and secondary contact zones often occur at ecotones between major associations of habitat and climate. Therefore, understanding processes that limit sympatry between species in such areas provides an important framework for testing biogeographic and evolutionary hypotheses. Theoretical and empirical work has shown that the evolution of species borders is influenced by a complexity of factors, including gene flow from central to peripheral populations and the ability of species to adapt locally to environmental conditions. However, few studies have used bioclimatic models, combined with molecular and morphological data, to predict geographic range limits in the context of gene flow across a secondary contact zone. In this study, I applied these methods to test specific hypotheses about barriers to sympatry between closely related species where they approach and contact each other. Specifically, I examined the importance of historical isolation, local adaptation, and symmetry of gene flow in limiting sympatry and range expansion of ecologically distinct species across environmental gradients. Molecular (mitochondrial DNA, allozymes), morphological, and bioclimatic data were obtained for two avian sibling species (Baeolophus inornatus and B. ridgwayi) that exist in recent, narrow secondary contact in northern California. These species are broadly allopatric and occupy rangewide associations of oak and pinyon-juniper woodlands, respectively, although B. inornatus also inhabits mixed or juniper woodlands locally. Patterns of molecular variation generally were congruent with morphological and bioclimatic data, and support prior evidence for a history of isolation, adaptation, and divergence in distinctive, species-specific vegetation-climate associations. However, molecular and morphological clines fall east of the limit of oaks, and individuals of B. inornatus in this juniper-associated contact zone experience bioclimates that are more similar to B. ridgwayi than to B. inornatus in oak habitat. Thus, B. inornatus is able to adapt and expand locally into the range of its close relative, but not vice versa. These data support the hypothesis that gene flow is asymmetrical where peripheral populations meet at range boundaries. Physiological differences between species may play an important role in influencing these patterns. Empirical studies that highlight the importance of local adaptation and patterns of gene flow in which closely related species contact across ecotones are central to understanding limits on geographic ranges, sympatry, and introgression-a cornerstone of biogeographic and speciation theory. PMID- 15341160 TI - Convergent evolution of Darwin's finches caused by introgressive hybridization and selection. AB - Between 1973 and 2003 mean morphological features of the cactus finch, Geospiza scandens, and the medium ground finch, G. fortis, populations on the Galapagos island of Daphne Major were subject to fluctuating directional selection. An increase in bluntness or robustness in the beak of G. scandens after 1990 can only partly be explained by selection. We use 16 microsatellite loci to test predictions of the previously proposed hypothesis that introgressive hybridization contributed to the trend, resulting in genes flowing predominantly from G. fortis to G. scandens. To identify F1 hybrids and backcrosses we use pedigrees where known, supplemented by the results of assignment tests based on 14 autosomal loci when parents were not known. We analyze changes in morphology and allelic composition in the two populations over a period of 15-20 years. With samples that included F1 hybrids and backcrosses, the G. scandens population became more similar to the G. fortis population both genetically and morphologically. Gene flow between species was estimated to be three times greater from G. fortis to G. scandens than in the opposite direction, resulting in a 20% reduction in the genetic difference between the species. Nevertheless, removing identified F1 hybrids and backcrosses from the total sample and reanalyzing the traits did not eliminate the convergence. The two species also converged in beak shape by 22.2% and in body size by 45.5%. A combination of introgressive hybridization and selection jointly provide the best explanation of convergence in morphology and genetic constitution under the changed ecological conditions following a major El Nino event in 1983. The study illustrates how species without postmating barriers to gene exchange can alternate between convergence and divergence when environmental conditions oscillate. PMID- 15341161 TI - Adaptive size modification by dominant female meerkats. AB - In species of cooperative insects that live in large groups, selection for increased fecundity has led to the evolution of an increased body size among female reproductives, but whether this is also true of cooperative vertebrates is unknown. Among vertebrates, morphological modification of female breeders has only been documented in a single species; in naked mole rats (Heterocephalus glaber), acquisition of alpha status is associated with a significant increase in body size through an elongation of the lumbar vertebrae. Here we provide evidence of morphological modification among breeding females of a cooperative carnivore, the meerkat (Suricata suricatta), and demonstrate that this modification is likely to be adaptive. The same female meerkats were significantly larger when they were dominant than when they were subordinate. This increased body size was not explained by differences in age, foraging efficiency, or investment in offspring care, but may have arisen, in part, through increased levels of hormone that govern bone growth. Increases in body size are likely to result in fitness benefits, for large females delivered larger litters and had heavier offspring, both of which are known to correlate positively with measures of breeding success in meerkats. Our results suggest that the acquisition of alpha status in female meerkats is associated with an adaptive increase in body size and hence that morphological modification of female vertebrates may be more widespread than has been previously supposed. PMID- 15341162 TI - Individual-level selection as a cause of Cope's rule of phyletic size increase. AB - Cope's rule, the tendency for species within a lineage to evolve towards larger body size, has been widely reported in the fossil record, but the mechanisms leading to such phyletic size increase remain unclear. Here we show that selection acting on individual organisms generally favors larger body size. We performed an analysis of the strength of directional selection on size compared with other quantitative traits by evaluating 854 selection estimates from 42 studies of contemporaneous natural populations. For size, more than 79% of selection estimates exceed zero, whereas for other morphological traits positive and negative values are similar in frequency. The selective advantage of increased size occurs for traits implicated in both natural selection (e.g., differences in survival) and sexual selection (e.g., differences in mating success). The predominance of positive directional selection on size within populations could translate into a macroevolutionary trend toward increased size and thereby explain Cope's rule. PMID- 15341163 TI - Estimating the strength of sexual selection from Y-chromosome and mitochondrial DNA diversity. AB - We show that a sex difference in the opportunity for selection results in sex differences in the strength of random genetic drift and thus creates different patterns of genetic diversity for maternally and paternally inherited haploid genes. We derive the effective population size Ne for a male-limited or female limited haploid gene in terms of I, the "opportunity for selection" or the variance in relative fitness. Because the variance in relative fitness of males can be an order of magnitude larger than that of females, the Ne is much smaller for males than it is for females. We derive both nonequilibrium and equilibrium expressions for F(ST) in terms of I and show how the portion of I owing to sexual selection, Imates, that is, the variation among males in mate numbers, is a simple function of the F's for cytoplasmic (female inherited) and Y-linked (male inherited) genes. Because multiple, transgenerational data are lacking to apply the nonequilibrium expression, we apply only the equilibrium model to published data on Y chromosome and mitochondrial sequence divergence in Homo sapiens to quantify the opportunity for sexual selection. The estimate suggests that sexual selection in humans represents a minimum of 54.8% of total selection, supporting Darwin's proposal that sexual selection has played a significant role in human evolution and the recent proposal regarding a shift from polygamy to monogamy in humans. PMID- 15341164 TI - Linkage between nuclear and mitochondrial DNA sequences in avian malaria parasites: multiple cases of cryptic speciation? AB - Analyses of mitochondrial cytochrome b diversity among avian blood parasites of the genera Haemoproteus and Plasmodium suggest that there might be as many lineages of parasites as there are species of birds. This is in sharp contrast to the approximately 175 parasite species described by traditional methods based on morphology using light microscopy. Until now it has not been clear to what extent parasite mitochondrial DNA lineage diversity reflects intra- or interspecific variation. We have sequenced part of a fast-evolving nuclear gene, dihydrofolate reductase-thymidylate synthase (DHFR-TS), and demonstrate that most of the parasite mitochondrial DNA lineages are associated with unique gene copies at this locus. Although these parasite lineages sometimes coexist in the same host individual, they apparently do not recombine and could therefore be considered as functionally distinct evolutionary entities, with independent evolutionary potential. Studies examining parasite virulence and host immune systems must consider this remarkable diversity of avian malaria parasites. PMID- 15341165 TI - Sperm survival in female stalk-eyed flies depends on seminal fluid and meiotic drive. AB - Sperm competition is common in many insect species; however, the mechanisms underlying differences in sperm precedence are not well understood. In the stalk eyed fly, Cyrtodiopsis whitei (Diptera, Diopsidae), sperm precedence is influenced by the presence of sex chromosome meiotic drive. When drive-carrying males compete with non-driving males for fertilizations within a female, the number of progeny sired by drive males is significantly fewer than predicted by sperm mixing alone. Thus, drive males apparently suffer not only a reduction in the number of viable sperm produced, but also a reduction in sperm competitive ability. In this study, we manipulated the amount and source of seminal fluid and sperm received by females by interrupting copulations before sperm, but after seminal fluid, was transferred. We find that seminal fluid from another male influences the number of progeny sired by a drive-carrying male when both males mate with the same female. Sperm viability staining reveals that sperm from drive males are incapacitated by seminal fluid from other males within the female reproductive tract. These results suggest that multiple mating by females enables seminal fluid products to interact differentially with sperm and may reduce the transmission advantage of the drive chromosome. PMID- 15341166 TI - Effects of developmental environment on signal-preference coupling in a Hawaiian cricket. AB - Previous work has suggested that developmental temperature influences expression of the adult male calling song of the cricket, a sexually selected mate recognition signal. The role of developmental temperature in shaping female preference functions, and thus its influence on signal-preference coupling has not been investigated. In this study, the effects of developmental temperature are examined in both males and females of the Hawaiian cricket, Laupala cerasina, to determine the degree of signal-preference matching between male song and female preference due to developmental environment. We found that rearing females in different temperature environments affected adult female acoustic preference functions in the same direction as male calling song, thereby influencing variation in adult reproductive behavior in such a way that male and female components remain coordinated. However, we further demonstrate that for male song, the effect of the rearing environment is not permanent but appears only to exert influence for a period of days. This mid-term temperature effect is distinct from the effect of short-term ambient temperature, which influences song in a matter of minutes and has been well documented. Signal-preference coordination, and sexual selection pressures due to mismatching within natural populations, likely will be influenced by nymphal developmental environments of males and females, as well as by adult singing and preference environments. PMID- 15341168 TI - Creep does not contribute to fatigue in bovine trabecular bone. AB - In both cortical and trabecular bone loaded in fatigue, the stress-strain loops translate along the strain axis. Previous studies have suggested that this translation is the result of creep associated with the mean stress applied in the fatigue test. In this study, we measured the residual strrain (corresponding to the translation of the stress-strain loops) in fatigue tests on bovine trabecular bone and compared it to an upper bound estimate of the creep strain in each test. Our results indicate that the contribution of creep to the translation of the stress-strain loops is negligible in bovine trabecular bone. These results, combined with models for fatigue in lower density bone, suggest that that creep does not contribute to the fatigue of normal human bone. Creep may make a significant contribution to fatigue in low-density osteoporotic bone in which trabeculae have resorbed, reducing the connectivity of the trabecular structure. PMID- 15341169 TI - A phenomenological model for predicting fatigue life in bovine trabecular bone. AB - Cyclic loading of bone during daily activities can lead to fatigue degradation and increased risk of fracture in both the young and elderly population. Damage processes under cyclic loading in trabecular bone result in the reduction of the elastic modulus and accumulation of residual strain. These effects increase with increasing stress levels, leading to a progressive reduction in fatigue life. The present work analyzes the effect of stress and strain variation on the above damage processes in bovine trabecular bone, and develops a phenomenological model relating fatigue life to the imposed stress level. The elastic modulus reduction of the bone specimens was observed to depend on the maximum compressive strain, while the rate of residual strain accumulation was a function of the stress level. A model was developed for the upper and lower bounds of bone elastic modulus reduction with increasing number of cycles, at each stress range. The experimental observations were described well by the model. The model predicted the bounds of the fatigue life with change in fatigue stress. The decrease in the fatigue life with increasing stress was related to corresponding increases in the residual strain accumulation rates at the elevated stress levels. The model shows the validity of fatigue predictions from relatively few cyclic experiments, by combining trends observed in the monotonic and the cyclic tests. The model also presents a relatively simple procedure for predicting the endurance limit for bovine trabecular bone specimens. PMID- 15341170 TI - A molecular rotor as viscosity sensor in aqueous colloid solutions. AB - BACKGROUND: Molecular rotors exhibit viscosity-dependent quantum yield, allowing non-mechanical determination of fluid viscosity. We analyzed fluorescence in the presence of viscosity-modulating macromolecules several orders of magnitude larger than the rotor molecule. METHOD OF APPROACH: Fluorescence of aqueous starch solutions with a molecular rotor in solution was related to viscosity obtained in a cone-and-plate viscometer. RESULTS: In dextran solutions, emission intensity was found to follow a power-law relationship with viscosity. Fluorescence in hydroxyethylstarch solutions showed biexponential behavior with different exponents at viscosities above and below 1.5 mPa s. Quantum yield was generally higher in hydroxyethylstarch than in dextran solutions. The power-law relationship was used to backcalculate viscosity from intensity with an average precision of 2.2% (range of -5.5% to 5.1%). CONCLUSIONS: This study indicates that hydrophilic molecular rotors are suitable as colloid solution viscosity probes after colloid-dependent calibration. PMID- 15341171 TI - Flow-induced endothelial surface reorganization and minimization of entropy generation rate. AB - Effects of hydrodynamic shear on the shape of the endothelial surface are examined based on evaluations of the rate of entropy generation at the cell surface. A linear solution of the flow over a sinusoidally varying endothelial surface is used to evaluate the entropy generation rate on the cell surface for which measured cell dimensions are available. Both the local rate of entropy generation (equivalent to the rate of energy dissipation by viscous shear) at the peak of a cell and the total entropy generation rate over the cell surface are minimized under conditions of a constant cell surface area and a constant cell peak height; which yields horizontal cell dimensions that are close to those obtained experimentally. PMID- 15341172 TI - Simultaneous in vitro measurement of patellofemoral kinematics and forces. AB - This study involved the development and testing of a system for the simultaneous in vitro measurement of tibiofemoral kinematics and patellofemoral kinematics and forces. Knee motion was tracked using a Vicon 370, and patellofemoral force was measured using a six degree-of-freedom transducer based on the design of Singerman et al. Using this system, twelve knee specimens were tested in supine leg extension under a simulated quadriceps force. The comprehensive set of results corresponds well to the individual results of previous studies. The measurement system will be of value in assessing the effects of total knee arthroplasty on patellar function. PMID- 15341173 TI - Measuring alignment of the hindfoot. AB - In subtalar arthrodesis operations, correction of the hindfoot alignment is performed in about half of the cases. To improve the quality of the operation, a measurement system was developed which reliably measures the hindfoot angle pre-, per-, and postoperatively. This device was evaluated by measuring subjects in standing weightbearing position and in prone nonweightbearing position. The results were compared with hindfoot angles constructed on posterior photographic images. The results are similar to other studies (all maximum values): intratester accuracy 1.4 degrees, intertester accuracy 2.2 degrees, intratester reliability 0.9, and intertester reliability 0.74. The proposed device will improve the quality of correction, because it enables peroperative measurement of hindfoot alignment. PMID- 15341174 TI - Effect of a lipid pool on stress/strain distributions in stenotic arteries: 3-D fluid-structure interactions (FSI) models. AB - Nonlinear 3-D models with fluid-structure interactions (FSI) based on in vitro experiments are introduced and solved by ADINA to perform flow and stress/strain analysis for stenotic arteries with lipid cores. Navier-Stokes equations are used as the governing equations for the fluid. Hyperelastic Mooney-Rivlin models are used for both the arteries and lipid cores. Our results indicate that critical plaque stress/strain conditions are affected considerably by stenosis severity, eccentricity, lipid pool size, shape and position, plaque cap thickness, axial stretch, pressure, and fluid-structure interactions, and may be used for possible plaque rupture predictions. PMID- 15341175 TI - A 2-D model of flow-induced alterations in the geometry, structure, and properties of carotid arteries. AB - Evidence from diverse investigations suggests that arterial growth and remodeling correlates well with changes in mechanical stresses from their homeostatic values. Ultimately, therefore, there is a need for a comprehensive theory that accounts for changes in the 3-D distribution of stress within the arterial wall, including residual stress, and its relation to the mechanisms of mechanotransduction. Here, however, we consider a simpler theory that allows competing hypotheses to be tested easily, that can provide guidance in the development of a 3-D theory, and that may be useful in modeling solid-fluid interactions and interpreting clinical data. Specifically, we present a 2-D constrained mixture model for the adaptation of a cylindrical artery in response to a sustained alteration in flow. Using a rule-of-mixtures model for the stress response and first order kinetics for the production and removal of the three primary load-bearing constituents within the wall, we illustrate capabilities of the model by comparing responses given complete versus negligible turnover of elastin. Findings suggest that biological constraints may result in suboptimal adaptations, consistent with reported observations. To build upon this finding, however, there is a need for significantly more data to guide the hypothesis testing as well as the formulation of specific constitutive relations within the model. PMID- 15341176 TI - Finite-element modeling of the hemodynamics of stented aneurysms. AB - BACKGROUND: Computational fluid dynamics (CFD) simulations are used to analyze the wall shear stress distribution and flow streamlines near the throat of a stented basilar side-wall aneurysm. Previous studies of stented aneurysm flows used low mesh resolution, did not include mesh convergence analyses, and depended upon conformal meshing techniques that apply only to very artificial stent geometries. METHOD OF APPROACH: We utilize general-purpose computer assisted design and unstructured mesh generation tools that apply in principle to stents and vasculature of arbitrary complexity. A mesh convergence analysis for stented steady flow is performed, varying node spacing near the stent. Physiologically realistic pulsatile simulations are then performed using the converged mesh. RESULTS: Artifact-free resolution of the wall shear stress field on stent wires requires a node spacing of approximately 1/3 wire radius. Large-scale flow features tied to the velocity field are, however, captured at coarser resolution (nodes spaced by about one wire radius or more). CONCLUSIONS: Results are consistent with previous work, but our methods yield more detailed insights into the complex flow dynamics. However, routine applications of CFD to anatomically realistic cases still depend upon further development of dedicated algorithms, most crucially to handle geometry definition and mesh generation for complicated stent deployments. PMID- 15341177 TI - Geometric determinants to cement line debonding and osteonal lamellae failure in osteon pushout tests. AB - Cement lines are the boundaries between secondary osteons and the surrounding interstitial bone matrix in cortical bone. The interfacial properties of cement lines have been determined by osteon pushout tests. However, distinctively different material properties were obtained when osteon pushout tests were performed under different test geometries. In the present study, an axisymmetric two-dimensional finite element model was used to simulate an osteon pushout test using the test geometry of actual experiments. The results indicated that shear failure within the osteonal lamellae would occur when the osteon pushout test was performed under the condition of a thick specimen and large supporting hole. On the other hand, cement line debonding occurred when the osteon pushout test was performed using a thin specimen and small supporting hole. The finite element results were consistent with previous experiments of osteon pushout tests under different test geometries. Furthermore, the finite-element results suggest that a smoothly curved punch would most likely cause debonding at the cement line instead of osteonal lamellae. PMID- 15341178 TI - Herb, nutrient and drug interactions may be hazardous. PMID- 15341179 TI - New JCAHO standards for pain management. PMID- 15341180 TI - The attraction of magnet nursing services. PMID- 15341181 TI - Nitric oxide and cellular stress response in brain aging and neurodegenerative disorders: the role of vitagenes. AB - Nitric oxide and other reactive nitrogen species appear to play crucial roles in the brain such as neuromodulation, neurotransmission and synaptic plasticity, but are also involved in pathological processes such as neurodegeneration and neuroinflammation. Acute and chronic inflammation result in increased nitrogen monoxide formation and nitrosative stress. It is now well documented that NO and its toxic metabolite, peroxynitrite, can inhibit components of the mitochondrial respiratory chain leading to cellular energy deficiency and, eventually, to cell death. Within the brain, the susceptibility of different brain cell types to NO and peroxynitrite exposure may be dependent on factors such as the intracellular reduced glutathione and cellular stress resistance signal pathways. Thus neurons, in contrast to astrocytes, appear particularly vulnerable to the effect of nitrosative stress. Evidence is now available to support this scenario for neurological disorders such as Alzheimer's disease, amyotrophic lateral sclerosis, Parkinson's disease, multiple sclerosis and Huntington's disease, but also in the brain damage following ischemia and reperfusion, Down's syndrome and mitochondrial encephalopathies. To survive different types of injuries, brain cells have evolved integrated responses, the so-called longevity assurance processes, composed of several genes termed vitagenes and including, among others, members of the HSP system, such as HSP70 and HSP32, to detect and control diverse forms of stress. In particular, HSP32, also known as heme oxygenase-1 (HO 1), has received considerable attention, as it has been recently demonstrated that HO-1 induction, by generating the vasoactive molecule carbon monoxide and the potent antioxidant bilirubin, could represent a protective system potentially active against brain oxidative injury. Increasing evidence suggests that the HO-1 gene is redox-regulated and its expression appears closely related to conditions of oxidative and nitrosative stress. An amount of experimental evidence indicates that increased rate of free radical generation and decreased efficiency of the reparative/degradative mechanisms, such as proteolysis, are factors that primarily contribute to age-related elevation in the level of oxidative stress and brain damage. Given the broad cytoprotective properties of the heat shock response there is now strong interest in discovering and developing pharmacological agents capable of inducing such a response. These findings have led to new perspectives in medicine and pharmacology, as molecules inducing this defense mechanism appear to be possible candidates for novel, cytoprotective strategies. Particularly, manipulation of endogenous cellular defense mechanisms such as the heat shock response, through nutritional antioxidants or pharmacological compounds, represents an innovative approach to therapeutic intervention in diseases causing tissue damage, such as neurodegeneration. Consistent with this notion, maintenance or recovery of the activity of vitagenes may possibly delay the aging process and decrease the occurrence of age-related diseases with resulting prolongation of a healthy life span. PMID- 15341182 TI - Stimulation of nerve growth factor expression in astrocytes by peroxynitrite. AB - BACKGROUND: Overproduction of nitric oxide (NO) has been recognized as a major mechanism of neurotoxicity. NO reacts with superoxide to generate peroxynitrite, a strong oxidizing and nitrating species. Peroxynitrite is formed in glial cells and degenerating neurons in neuropathological conditions, including amyotrophic lateral sclerosis (ALS). In ALS, motor neurons re-express the p75 neurotrophin receptor (p75NTR) and might become vulnerable to NGF. In the present study, we investigated whether peroxynitrite stimulated nerve growth factor (NGF) expression in spinal cord astrocytes. MATERIALS AND METHODS: Astrocyte monolayers were exposed to peroxynitrite and nitrotyrosine formation was determined by immunofluorescence. mRNA levels for NGF, brain derived neutrophic factor (BDNF) and fibroblast growth factor-2 (FGF-2) were measured by semi-quantitative RT-PCR and NGF release was determined by ELISA. RESULTS AND DISCUSSION: A single exposure to peroxynitrite specifically induced NGF expression and secretion in astrocytes coincident with reactive morphological changes and increased nitrotyrosine immunoreactivity. These results suggest that NGF expression in reactive astrocytes is under the control of oxidative stress. PMID- 15341183 TI - Astrocytic nitric oxide triggers tau hyperphosphorylation in hippocampal neurons. AB - Production of nitric oxide (NO) by glial cells has been proposed to mediate cytotoxic effects on neighboring neurons. Although extensive genetic data implicate the beta amyloid peptide (Abeta) in the neurodegenerative cascade of Alzheimer's disease (AD), the molecular mechanisms underlying its effects on neurons and glia and the relationship between glial activation and neuronal death are not well understood. In AD, Abeta is sufficient to induce glial activation and promote the generation of inflammatory mediators including NO. We examined whether Abeta stimulated astrocytes to express nitric oxide synthase and produce NO. Also, we investigated whether astrocytic NO contributes to degenerative changes occurring in co-cocultured hippocampal neurons. We found that the treatment of rat hippocampal astrocyte cultures with Abeta(25-35) fragment up regulated the mRNA and protein levels of both the inducible and neuronal forms of nitric oxide synthase (iNOS and nNOS, respectively) and increased the production of nitric oxide. Remarkably, hippocampal neurons co-cultured with astrocytes, previously stimulated with Abeta, displayed hyperphosphorylation of the microtubule-associated protein tau. This effect was attenuated by iNOS inhibitors, suggesting the role of overproduction of NO by reactive astrocytes in AD pathogenesis. PMID- 15341184 TI - The role of nitric oxide in the radiation-induced effects in the developing brain. AB - The immature and adult brain display clear differences in the way they respond to insults. The effects of prenatal irradiation on the developing brain are well known. Both epidemiological and experimental data indicate that ionizing radiation may disrupt developmental processes leading to deleterious effects on post-natal brain functions. A central role of reactive oxygen and nitrogen species (ROS/RNS) as important mediators in both neurotoxicity and neuroprotection has been demonstrated. However, data concerning the role of ROS/RNS in radiation-induced damage in the developing brain are scarce. The goal of this review was to summarize the current studies concerning the role of nitric oxide and its reactive intermediates in activation of signal transduction pathways involved in cellular radiation response, with particular focus on radiation-induced effects in the developing brain. PMID- 15341185 TI - Differential role of extra- and intracellular superoxide anions for nitric oxide mediated apoptosis induction. AB - Nitric oxide (NO) has recently been shown to mediate apoptosis induction selectively in transformed fibroblasts, in contrast to their nontransformed parental cells. Here we show that NO-mediated apoptosis induction in transformed fibroblasts can be divided into two major phases. During phase 1, peroxynitrite is generated by the interaction of extracellular superoxide anions with NO and the intracellular glutathione level is subsequently lowered. This defines the beginning of phase 2, in which NO-mediated signaling depends on intracellular superoxide anions exclusively. The resultant peroxynitrite seems to activate the mitochondrial permeability transition pore and thus triggers execution of apoptosis. Experimental depletion of intracellular glutathione causes a drastic decrease in the length of phase 1 in transformed cells and renders nontransformed cells sensitive to NO-mediated apoptosis induction. These findings allow the prediction that either induction of superoxide anion generation or glutathione depletion may render cells sensitive to NO-mediated apoptosis induction. PMID- 15341186 TI - Modulation of retinal dopaminergic cells by nitric oxide. A protective effect on NMDA-induced retinal injury. AB - Nitric oxide (NO) may play an important role in regulating retinal neuronal survival. While the precise impact of NO mechanisms on retinal neurons remains to be elucidated, it has been reported that low doses of NO may have a neuroprotective effect against N-methyl-D-aspartate (NMDA)-induced retinal neurotoxicity. Dopamine has also been recognized to have neuroprotective actions. Retinal dopaminergic cells can be detected by the immunohistochemical staining of tyrosine hydroxylase (TH), the rate-limiting enzyme in dopamine synthesis. We observed that NMDA dramatically decreased in TH immunostaining at the junction between the inner nuclear layer and inner plexiform layer, and that this reduction in immunostaining was attenuated by the co-injection of NOC 18, an NO donor. Thus, the current review focused on the NO-dopamine interactions in the retinal neuroprotection. PMID- 15341187 TI - CCK-8S systemic administration blocks the 7-nitroindazole-induced effects on the EEG of striatum and globus pallidus: a FFT analysis in the rat. AB - BACKGROUND: Nitric oxide (NO) and cholecystokinin (CCK) are involved in the modulation of several neurotransmitter systems in the basal ganglia, and a functional interaction between their modulatory effects could be hypothesised. MATERIALS AND METHODS: We studied the effects exerted by the administration of 7 nitroindazole (7-NI) (50 mg kg(-1) i.p.), a selective inhibitor of neuronal NO synthase, on the depth EEG activity of the striatum and of the globus pallidus in both not pre-treated and sulphated CCK octapeptide (CCK-SS)-treated (100 nM kg( 1) i.p.) rats. Striatal and pallidal depth EEG power spectra were examined by means of a Fast Fourier Transform analysis. RESULTS: Striatal depth recordings showed a marked increase of % power of slow standard rhythms after 7-NI systemic treatment. In contrast, pallidal recordings revealed an increase of % power of rapid standard rhythms after i.p. injection of 7-NI. The same modifications were not evidenced in CCK-8S pre-treated rats after 7-NI administration. CONCLUSION: The results show an influence exerted by peripheral CCK on the nitrergic modulation of the bioelectric activities of the striatum and of the globus pallidus. This effect could be of particular interest in the light of NO and CCK involvement in the neuroprotective mechanisms. PMID- 15341188 TI - The role of nitric oxide in the pathogenesis of brain lesions during the development of Alzheimer's disease. AB - Nitric oxide (NO) is a key bioregulatory active molecule in the cardiovascular, immune and nervous systems, synthesized through converting L-arginine to L citrulline by NO synthase (NOS). Research exploration supports the theory that this molecule appears to be one of the key factors for the disruption of normal brain homeostasis, which causes the development of brain lesions and pathology such as in Alzheimer's disease (AD). Especially the vascular content of NO activity appears to be a major contributor to this pathology before the overexpression of NOS activity in other brain cellullar compartments develop. We theorize that pharmacological intervention using NO donors and/or NO suppressors should delay or minimize brain lesion development and further progression of brain pathology and dementia. PMID- 15341189 TI - Apoptosis and cytochrome c release in cerebellar granule cells. AB - In the light of both the major role played by released cytochrome c in apoptosis of a variety of cells and the availability of cerebellar granule cells as a model system to investigate apoptosis as a function of time from induction to cell death, we review data aimed at elucidating the events dealing with cytochrome c release from mitochondria as well as its role outside mitochondria. We report cytochrome c release in the apoptosis time course as dependent on the function of both the antioxidant and proteolytic systems. We show that, beside the role played by cytochrome c in participating in apoptosome formation and in triggering the caspase cascade, at least in cerebellar granule cells, released cytochrome c can maintain its ability to work as an electron carrier, being a scavenger of reactive oxygen species and an electron donor to cytochrome oxidase, thus driving the ATP synthesis. PMID- 15341190 TI - Functional significance of nitric oxide in the inner ear. AB - Significant advances have been made in our understanding of the functional significance of nitric oxide (NO) in the inner ear. The localization of NO synthase and the nitric oxide production site has now been established by immunohistochemistry and the fluorescent indicator of NO. The functional significance of NO in the inner ear, especially as a neurotransmitter, is becoming increasingly clear. Mounting evidence suggests that excessive NO production may play an essential role in inner ear disorders as well. The production of an inducible type of NO synthase may be closely related to this phenomenon. Based on the mechanisms of inner ear disorders, new pharmacological strategies for preventing and/or treating inner ear disorders have also been suggested. PMID- 15341191 TI - A potent antioxidant, lycopene, affords neuroprotection against microglia activation and focal cerebral ischemia in rats. AB - We investigated the effects of a potent antioxidant, lycopene, on the free radical-scavenging activity as evaluated by the DPPH test and lipid peroxidation in rat brain homogenates as well as nitric oxide (NO) formation in cultured microglia stimulated by lipopolysaccharide. In addition, we also investigated the therapeutic effect of lycopene in attenuating ischemia/reperfusion brain injury induced by middle cerebral artery (MCA) occlusion in rats. Lycopene (1, 2 and 5 microM) exerted increased DPPH decolorization in the DPPH test, and increased inhibition of iron-catalyzed lipid peroxidation (TBARS formation) in rat brain homogenates in concentration-dependent manners. Furthermore, lycopene (5 and 10 microM) significantly inhibited nitrite production by about 31% and 61% in microglia stimulated by LPS, respectively. Rats which received lycopene at a dosage of 4 mg/kg, but not at 2 mg/kg, showed significant infarct size reductions compared with those which received the solvent control (20% Tween 80). In conclusion, we demonstrate a protective effect of lycopene on ischemic brain injury in vivo. Lycopene, through its antioxidative property, mediates at least a portion of free radical-scavenging activity and inhibits microglia activation, resulting in a reduction in infarct volume in ischemia/reperfusion brain injury. PMID- 15341192 TI - Nitric oxide and brain hyperexcitability. AB - Nitric oxide (NO) is a gaseous messenger involved in atypical forms of intercellular communications, able to exert a strong functional modulation of several neurotransmitter systems. In particular, NO heavily influences the excitatory neurotransmitter glutamate, mainly through NMDA receptors, and the inhibitory neurotransmitter GABA, mainly through GABA A receptors. Due to the involvement of glutamate and GABA in a delicate balance conditioning the functional status of the neural cells, this interaction suggests a role for NO in regulating neuronal excitability and its transition towards hyperexcitability phenomena. This article reviews the main knowledge about the relationships existing between the activity of the NO system and the experimental aspects of epilepsy, focusing on the somewhat antithetic findings about the proconvulsant or the anticonvulsant roles exerted by nitric oxide. PMID- 15341193 TI - Regulation of programmed cell death in neuronal cells by nitric oxide. AB - Nitric oxide (NO), produced from L-arginine and molecular oxygen in a reaction catalyzed by one of three NO synthase isoenzymes, can prevent or induce neuronal apoptosis depending on its concentration and cellular redox state. This molecule affords neuroprotection by post-translational S-nitrosylation of NMDA receptor, caspases and p21ras, and increases the expression of cytoprotective genes such as HSP70, heme oxygenase and Bcl-2. Moreover, the NO/cGMP pathway activates the anti apoptotic serine/threonine kinase Akt by protein kinase G-dependent activation of phosphatidylinositol 3-kinase. A high concentration of NO and peroxynitrite, a reaction product of NO with superoxide anion, can promote apoptotic pathways in neuronal cells through the indirect activation of caspases. We review the molecular mechanism by which NO exerts both pro- and anti-apoptotic actions in neuronal cells and the clinical implications for regulating neuronal apoptosis. PMID- 15341194 TI - Nitric oxide as a physiopathological factor in neuropsychiatric disorders. AB - The dominant research subject on schizophrenia, mood disorders, autism and other central nervous system diseases has been related to neurotransmitter system abnormalities. For example, the dopamine hypothesis states that schizophrenia is the result of dopaminergic hyperactivity. The therapeutic approach has also been directed towards finding agents which will modulate or regulate these neurotransmitter systems at any step. There is substantial and mounting evidence that subtle abnormalities of reactive oxygen species (ROS) and nitric oxide (NO) may underlie a wide range of neuropsychiatric disorders. NO has chemical properties that make it uniquely suitable as an intracellular and intercellular messenger. It is produced by the activity of nitric oxide synthases which are present in peripheral tissues and in neurons. On the other hand, NO is known to be an oxygen radical in the central and peripheral nervous systems. NO has been implicated in a number of physiological functions such as noradrenaline and dopamine releases, memory and learning and certain pathologies such as schizophrenia, bipolar disorder and major depression. Evidence has been considered here for the proposal that an abnormality of NO metabolism may be a contributory factor in some neuropsychiatric disorders. The direct evidence for NO abnormalities in schizophrenia and other psychiatric disorders remains relatively limited to date, although there are some clinical and experimental studies. The suggestion that NO and other ROS may play a role in some neuropsychiatric disorders clearly has important implications for new treatment possibilities. The primary objective of the present review was to summarize and critically evaluate the current knowledge regarding a potential contribution of NO to the neuropathophysiology of schizophrenia as well as other neuropsychiatric disorders. PMID- 15341195 TI - In vitro evidence supporting the therapeutic role of nicotine against neurodegeneration. AB - This review supports the necessity of combining fundamental chemical and biological methods to scrutinize potential causative agents in neurodegeneration. This is supported by recent experimental evidence in relation to the use of nicotine as a potential therapeutic agent, especially when following the path of iron's role in catalysing the generation of reactive oxygen species via a Fenton like reaction. Exploration of the dose-response relationship indicates that acute administration offers the most likely success, reducing tremor and improving cognitive performance amongst others. Confirmation of this relationship is gathered from recent in vivo and in vitro efforts that support this hypothesis. PMID- 15341196 TI - Early development of children at familial risk for dyslexia--follow-up from birth to school age. AB - We review the main findings of the Jyvaskyla Longitudinal study of Dyslexia (JLD) which follows the development of children at familial risk for dyslexia (N = 107) and their controls (N = 93). We will illustrate the development of these two groups of children at ages from birth to school entry in the skill domains that have been connected to reading and reading disability in the prior literature. At school entry, the highest score on the decoding task among the poorer half (median) of the at risk children--i.e. of those presumably being most likely genetically affected--is 1 SD below the mean of the control group. Thus, the familial risk for dyslexia shows expected consequences. Among the earliest measures in which group differences as well as significant predictive associations with the first steps in reading have emerged, are indices of speech processing in infancy. Likewise, various measures of early language including pronunciation accuracy, phonological, and morphological skills (but not performance IQ) show both group differences and predictive correlations, the majority of which become stronger as the reliability of the measures increases by age. Predictive relationships tend to be strong in general but higher in the at risk group because of its larger variance in both the predictor variables and in the dependent measures, such as early acquisition of reading. The results are thus promising in increasing our understanding needed for early identification and prevention of dyslexia. PMID- 15341197 TI - The early identification of dyslexia: children with English as an additional language. AB - It is generally accepted that dyslexia should be identified early for interventions to have maximum effect. However, when children speak English as an additional language (EAL), diagnosis is more complex and there is concern that these children tend to be under-identified. This paper reports a longitudinal study following the development of phonological awareness skills in relation to progress in learning to read with a cohort of British Asian children learning EAL and their monolingual peers. It also sought to determine the usefulness of a measure of phonological skills for the identification of dyslexic-type difficulties in children learning EAL. Analysis revealed that both cohorts achieved similar levels of reading accuracy in school Years 2, 4 and 6, with higher levels of reading comprehension for the monolingual children and faster reading fluency for children learning EAL in each school year. There was a similar pattern of relationships between the reading measures and measures of phonological awareness for both groups of children. However, monolingual children achieved higher levels of rhyme detection and alliteration fluency whilst the children learning EAL achieved faster number naming times. Overall, a phonological assessment battery was useful in identifying reading accuracy related difficulties in both groups of children. However, concerns are raised about the sensitivity of such measures following the introduction of the Literacy Hour. PMID- 15341198 TI - Working memory: its role in dyslexia and other specific learning difficulties. AB - This paper reports a study contrasting dyslexic children against a control group of children without special educational needs (SEN) and a group with varied SENs. Children's abilities were compared on tasks assessing phonological processing, visuo-spatial/motor coordination and executive/inhibitory functioning; being targeted for assessment based on theoretical proposals related to the working memory model. Primary and secondary school level children were tested: 21 assessed as dyslexic with no comorbid difficulties, 26 children assessed with difficulties including dyspraxia, emotional/behavioural problems and attention deficits, 40 children with no known education-related deficits were controls. Results indicated both SEN groups performed worse than controls on working memory phonological loop measures. However, SEN groups could only be differentiated on phonological awareness measures: the dyslexics showing lower scores. Dyslexics performed as well as controls on working memory visuo-spatial scratch pad measures and one of two additional visual-motor coordination tasks, whereas the performance of the other SEN children was lowest on the majority of these measures. Central executive and interference measures engendered mixed performances, both SEN groups showing evidence of deficits in one or more of these areas of functioning, although, of the two SEN groups, the dyslexics seem to have performed the worse when digit name processing was required. PMID- 15341199 TI - Auditory processing skills and phonological representation in dyslexic children. AB - It is now well-established that there is a causal connection between children's phonological skills and their acquisition of reading and spelling. Here we study low-level auditory processes that may underpin the development of phonological representations in children. Dyslexic and control children were given a battery of phonological tasks, reading and spelling tasks and auditory processing tasks. Potential relations between deficits in dyslexic performance in the auditory processing tasks and phonological awareness were explored. It was found that individual differences in auditory tasks requiring amplitude envelope rise time processing explained significant variance in phonological processing. It is argued that developmentally, amplitude envelope cues may be primary in establishing well-specified phonological representations, as these cues should yield important rhythmic and syllable-level information about speech. PMID- 15341200 TI - Visual and auditory temporal processing and early reading development. AB - This study investigated the ability of temporal processing measures obtained before school entry to predict early reading development in an unselected sample of 125 children (68 males, 57 females). Visual and auditory temporal order judgement (TOJ) tasks measured at Preschool (mean age 5.36 years) significantly predicted letter and word identification (accuracy) and reading rate (fluency) in early Grade 1 (mean age 5.94 years), even after the effects of age, environment, memory, attention, nonverbal ability, and speech/language problems were accounted for. There were no significant differences in the overall variance accounted for in reading between TOJ measures taken before or after reading had emerged. Both Preschool and Grade 1 measures of auditory TOJ accounted for significant independent variance in reading. However, only visual TOJ performance measured at Grade 1 accounted for unique variance in reading rate. This was discussed in terms of developmental changes in the role of visual temporal processing as reading develops. Reliability of the temporal measures from Preschool to Grade 1 was moderate. The results showed that measures of visual and auditory temporal processing obtained close to school-entry would be a useful addition to predicting risk of early reading difficulties. PMID- 15341201 TI - The Durham experience: promoting dyslexia and dyspraxia friendly schools. AB - The Durham County Policy Document on Specific Learning Difficulties (dyslexia and dyspraxia) has been developed over a period of years as a countywide response to ensure that children's specific learning difficulties are identified and that schools are equipped to offer a range of appropriate interventions. The content of the policy document is studied in depth along with the training programmes offered by the Learning Support Service and Educational Psychology Service that will support schools in putting the policy into practice. Future developments, including an exciting project on promoting inclusive practice for pupils with Specific Learning Difficulties (dyslexia and dyspraxia), are outlined. PMID- 15341202 TI - The age dynamics of monoamine oxidase activity and levels of lipid peroxidation products in the human brain. PMID- 15341203 TI - Intracortical synchronization of epileptic discharges at different stages of ultrastructural rearrangements in a completely neuronally isolated area of rat neocortex. AB - Cross-correlation functions were used to study the time delay (a measure of synchronization) in the appearance of epileptic discharges in distant areas of the cortex in the intact cortex and in neuronally isolated cortical strips in Wistar rats. Experiments were performed at different stages of axon sprouting 30 and 90 days after isolation of cortical areas and in intact cortex. Significant increases in the number of synapses in layer V of isolated cortical strips at 30 days correlated with significant decreases in the time delay, while decreases in the number of synapses at 90 days correlated with a significant increase in the time delay. This is evidence that newly formed synapses increase the extent of synchronization and thus affect epileptogenesis. The data obtained here suggest that large pyramidal cells in layer V of the rat neocortex form a neural network in pathological conditions, this supporting intracortical synchronization of epileptic discharges. PMID- 15341204 TI - Baseline spike activity of neurons in the locus coeruleus of the rat after lesions to a number of medulla oblongata nuclei. AB - Bilateral lesioning of the presublingual nucleus (nucleus praepositus hypoglossi) of the medulla oblongata induced significant changes in the nature of baseline spine activity in the locus coeruleus. After lesioning, the mean frequency of spike activity of locus coeruleus neurons decreased more than two-fold. The numbers of neurons with train-grouped activity and polymodal neurons increased significantly. Exclusion of the solitary tract nucleus led to an increase in the number of neurons with regular activity and some decrease in the mean discharge frequency in locus coeruleus neurons. These data support the suggestion that the presublingual nucleus plays a significant role in the transmission of afferent influences on the activity of locus coeruleus neurons. PMID- 15341205 TI - Local bicuculline disinhibition does not disrupt acquired coordination between the tactile input and the motor output in the motor cortex of the cat. PMID- 15341206 TI - An apparatus for recording defensive responses to tactile stimulation in the ground snail. AB - We describe here an original working experimental apparatus for non-invasive objective recording of the magnitudes of defensive responses to tactile stimulation in the ground snail. The apparatus includes a tracking device that ensures that the snail's position relative to a light and photodiode remains constant as it ascends a cylinder rotating about a horizontal axis. The device providing tactile stimulation is based on a loudspeaker circuit and provides blows of dosed energy. The snail responds to the tactile stimulation by retracting the antennae, head, and foot, which decreases the area of its shadow; this is recorded by a photodiode. PMID- 15341207 TI - Morphofunctional changes in Mauthner neurons during exposure to the neuropeptide kyotorphin. PMID- 15341208 TI - Comparative morphological analysis of neuron populations in the ganglion layer of the rodent cerebellum. PMID- 15341209 TI - Axon reactions precede demyelination in experimental models of multiple sclerosis. PMID- 15341210 TI - Cerebral cortex ultrastructure during exposure to hydrogen sulfide gas. PMID- 15341211 TI - Formation and structural organization of the barrier on the outer surface of the brain. PMID- 15341212 TI - Morphofunctional changes in adapted Mauthner neurons in goldfish after prolonged orthodromic stimulation of the auditory nerve in vitro. PMID- 15341213 TI - Characteristics of the surgical microneuroanatomy of formations of the posterior cranial fossa in relation to the retrosigmoidal and transcondylar approaches. PMID- 15341214 TI - Variability in the structure of field 39 of the lower parietal area of the cortex in the left and right hemispheres of adult human brains. AB - The cytoarchitectonics of field 39 of the lower parietal area of the cortex in the left and right hemispheres were studied in normal individuals and professionally gifted world-famous people. Series of frontal sections were studied, stained by the Nissl cresyl violet method. The profile area of neurons was measured in layers III and V and the distribution of neurons in terms of this property was analyzed. Gifted people showed a significantly greater level of individual variability in the measures recorded here, especially in the right hemisphere, with larger proportions of very small and very large neurons. PMID- 15341215 TI - Genetic determination of neurophysiological mechanisms of cortical-subcortical integration of bioelectrical brain activity. AB - The contribution of genetic factors to the formation of the neurophysiological mechanisms of cortical-subcortical integration was studied in 12 pairs of monozygotic and five pairs of dizygotic twins (aged 18-25 years). Intrapair similarity of the nature of spatial interactions between bioelectrical activity in the cerebral cortex, assessed from different combinations of statistical interactions of EEG from 16 monopolar recordings, was assessed in each pair of twins (and among 544 non-related pairs of subjects in both groups). The results suggest a high level of general population invariance and relatively small inherited and phenotypic variability in the morphofunctional systems making up the major neurophysiological mechanisms of brain integration as a whole. The ontogenetic formation of stem and subcortical regulatory structures, which have a leading role in the systems combination of different parts of the brain into a single formation, appears to occur in all individuals by the same principle, as disturbance can apparently affect the fundamental monomorphic features of the species. In turn, we might expect to find large interindividual variability in the establishment of interregional connections of the neocortex, the role of inherited and environmental factors being different in the processes forming long and relatively short intercortical interactions. PMID- 15341216 TI - Possible mechanisms of involvement of the amygdaloid complex in the control of gastric motor function. AB - Electrophysiological and neuroanatomical experiments on Wistar rats were performed to study the mechanisms of the modulatory influences of the amygdaloid nuclei on reflex motor activity in the stomach. Electrical stimulation of the central nucleus was accompanied by reproducible changes in the ongoing motor activity of the stomach in activity evoked by activation of the vagovagal reflex arc. The most marked, and predominantly inhibitory, effects were seen in response to stimulation of the medial part of the nucleus. Microinjections of the anterograde neuron marker Phaseolus vulgaris leucoagglutinin (PHA-L) into the central nucleus of the amygdala revealed the existence of direct descending projections from its dorsomedial part to the area containing cells of the vagosolitary complex, associated with performance of the vagovagal reflexes of the stomach. Electrical stimulation of this part of the central nucleus led to changes in neuron responses in the bulbar "gastric" center evoked by stimulation of the vagus nerve. These features may underlie one of the mechanisms of the amygdalar modulation of the reflex activity of the stomach. PMID- 15341217 TI - Characteristics of tremor in normal subjects and in the diagnosis and treatment of Parkinsonism. AB - Tensotremorography was used to record voluntary forces and to study the characteristics of the involuntary and voluntary components of isometrically recorded hand strength. The frequency ranges for changes in the spectral density of oscillations recorded here supported the existence of two suprasegmental systems associated with voluntary control and continuous regulation of force maintaining or holding a posture. Cross-correlation analysis of hand force maintained in conditions of visual feedback in normal conditions and in conditions of central disorders of the movement control system is presented. PMID- 15341218 TI - The neuroprotective effects of Semax in conditions of MPTP-induced lesions of the brain dopaminergic system. AB - This report describes studies cf the effects of the ACTH(4-10) analog Semax (MEHFPGP) on the behavior of white rats with lesions to the brain dopaminergic system induced by the neurotoxin MPTP. Neurotoxin was given as single i.p. doses of 25 mg/kg. Neurotoxin injections were shown to decrease movement activity and increase anxiety in the animals. Daily intranasal administration of Semax at a dose of 0.2 mg/kg decreased the severity of MPTP-induced behavioral disturbances. The protective activity of Semax in MPTP-induced lesions of the brain dopaminergic system may be associated with both its modulating effect on the dopaminergic system and the neurotrophic action of the peptide. PMID- 15341219 TI - Expression of the c-fos gene in spinal cord and brain cells in rats subjected to stress in conditions of exposure to various types of halothane anesthesia. AB - The influences of different treatments on the expression of the c-fos gene in the spinal cord and brain (hypothalamus) was studied in rats using various types of anesthesia. Synthesis of c-Fos-like proteins occurred only in the spinal cord in conditions of constant 1.5% halothane anesthesia. Use of induction anesthesia with 1.5% halothane allowed detection of c-Fos-like protein expression in cells of the rat spinal cord (lumbar segments) and brain, both when animals were placed in a hammock and when mechanical pain stimulation or electromagnetic irradiation of the skin with UHF currents were applied. The pattern of brain structures reacting to mechanical pain stimulation with expression of c-Fos-like protein was identified. This type of stimulation was shown to induce increases in the quantity of c-Fos-positive cells in the lateral hypothalamic area (LHA), the ventromedial (VMH) and dorsomedial (DMH) hypothalamic nuclei, and in the ventral hypothalamic area (AHA) by 116%, 167%, 101%, and 157% respectively as compared with controls. Skin irradiation with UHF currents decreased the intensity of mechanical pain stimulation-induced synthesis of c-Fos-like protein in most structures (LHA, VMH, DMN, and AHA by 32.8%, 29%, 15%, and 33% respectively). Only induction halothane anesthesia allowed identification of hypothalamic structures reacting to mechanical pain stimulation and the modifying effects of irradiating the skin with UHF currents on the intensity of these reactions. PMID- 15341220 TI - Correlation between cataleptic freezing and prepulse inhibition of the startle reflex in rats. AB - GC rats, bred for a predisposition to cataleptic freezing, were found to show a significant negative correlation between the duration of freezing and the level of prepulse inhibition of the startle reflex. In addition, a group of GC rats characterized by increased "nervousness" also showed a negative correlation between the duration of freezing and the extent of habituation of the startle reflex. These correlations were not seen in Wistar rats. Since decreases in the level of prepulse inhibition and habituation of the startle reflex are regarded as characteristic of schizophrenia, it is suggested that cataleptic freezing in GC rats might be used as a model of schizophrenic pyschopathology. PMID- 15341221 TI - Stimulation of the immune response during activation of the dopaminergic system in mice with opposite types of behavior. AB - Studies reported here demonstrated that activation of the dopaminergic system induces increases in the immune response regardless of the type of behavior in mice (line CBA), i.e., in aggressive mice, submissive mice, and mice lacking experience of victory or defeat (controls). Changes in the activity of the dopaminergic system were induced with SKF-38393, a selective agonist of dopamine D1 receptors, and with p-chlorophenylalanine (PCPA), which we have previously shown to activate D2 receptors. In the aggressive form of behavior, which was characterized by strong (compared with controls) immune responses, SKF-38393 and PCPA led to further increases in the immune response. In submissive mice, activation of the dopaminergic system altered the nature of the immune response, with immunostimulation, as in aggression. It is suggested that activation of the dopaminergic system in conditions of defined psychoemotional status fixed by acquisition of opposite types of behavior, induces the formation of a new neurochemical pattern--the dopaminergic set--which led to changes in the nature and intensity of the immune response. PMID- 15341222 TI - Cortical cognitive potentials in elderly persons. AB - The purpose of this study was to analyze the changes in the acoustically evoked cortical cognitive potentials N 200, P 300, and N 3 with age. There were 232 participants, who were 60 years or older, and each was examined using a battery of audiological tests including a questionnaire, otomicroscopy, pure-tone audiometry, and cortical cognitive potentials, measured at Fz using an auditory oddball paradigm. N 200 was elicited in 46.9%, P 300 in 45.1%, and N 3 in 52.2% of the elderly participants. The most significant predictors for presence of cortical responses were the participant's age and hearing level at target tone frequency. Monosyllabic speech recognition score was a less important predictor for presence of response. Response latency in the elderly sample increased steadily with age. Few changes in cortical response amplitude were found with age. We conclude that the speed of central auditory processing seems to be reduced with age. PMID- 15341223 TI - Newborn hearing screening with combined otoacoustic emissions and auditory brainstem responses. AB - Accurate assessment of neonatal hearing screening performance is impossible without knowledge of the true status of hearing, a prohibitive requirement that necessitates a complete diagnostic evaluation on all babies screened. The purpose of this study was to circumvent this limitation by integrating two types of screening measures obtained near simultaneously on every baby. Peripheral auditory function was defined by otoacoustic emission results. A complete diagnostic evaluation was performed on every baby who received a "Refer" outcome for auditory brainstem response screening. The integrated results for auditory brainstem response screening in an unselected group of 300 newborns estimated sensitivity at 100%, specificity at 99.7%, overall referral rate at 2.0%, and a positive predictive value of 83.3%. Conductive loss associated with amniotic fluid in the middle ear can persist several weeks after birth; conductive loss can produce a "Refer" outcome for auditory brainstem response screening; and auditory neuropathy can be detected with screening measures. Prevalence results were consistent with the published literature. The implications of this study are that otoacoustic emissions and auditory brainstem measures provide much more information than either alone and that both are needed for a comprehensive hearing screening program. PMID- 15341224 TI - Speech perception in noise: directional microphones versus frequency modulation (FM) systems. AB - The major consequence of sensorineural hearing loss (SNHL) is communicative difficulty, especially with the addition of noise and/or reverberation. The purpose of this investigation was to compare two types of technologies that have been shown to improve the speech-perception performance of individuals with SNHL: directional microphones and frequency modulation (FM) systems. Forty-six adult subjects with slight to severe SNHL served as subjects. Speech perception was assessed using the Hearing in Noise Test (HINT) with correlated diffuse noise under five different listening conditions. Results revealed that speech perception was significantly better with the use of the FM system over that of any of the hearing aid conditions, even with the use of the directional microphone. Additionally, speech perception was significantly better with the use of two hearing aids used in conjunction with two FM receivers rather than with just one FM receiver. Directional microphone performance was significantly better than omnidirectional microphone performance. All aided listening conditions were significantly better than the unaided listening condition. PMID- 15341225 TI - Performance of directional microphones for hearing aids: real-world versus simulation. AB - The purpose of this study was to assess the accuracy of clinical and laboratory measures of directional microphone benefit. Three methods of simulating a noisy restaurant listening situation ([1] a multimicrophone/multiloudspeaker simulation, the R-SPACE, [2] a single noise source behind the listener, and [3] a single noise source above the listener) were evaluated and compared to the "live" condition. Performance with three directional microphone systems differing in polar pattern (omnidirectional, supercardioid, and hypercardioid array) and directivity indices (0.34, 4.20, and 7.71) was assessed using a modified version of the Hearing in Noise Test (HINT). The evaluation revealed that the three microphones could be ordered with regard to the benefit obtained using any of the simulation techniques. However, the absolute performance obtained with each microphone type differed among simulations. Only the R-SPACE simulation yielded accurate estimates of the absolute performance of all three microphones in the live condition. Performance in the R-SPACE condition was not significantly different from performance in the "live restaurant" condition. Neither of the single noise source simulations provided accurate predictions of real-world (live) performance for all three microphones. PMID- 15341226 TI - Effects of discrimination task difficulty on N1 and P2 components of late auditory evoked potential. AB - The present study investigated the question of whether, in healthy young listeners, increases in discrimination task difficulty will alter the amplitude of either the N1 or P2 components of the late auditory evoked potential (LAEP). Using a stimulus oddball procedure, listeners discriminated changes in the frequency of ongoing tonal stimuli. On different test runs, task difficulty was manipulated by decreasing the size of the frequency differences and/or adding competing speech babble to the nontest ear. Both stimulus procedures produced significant decreases in P2 amplitude but had no effects on N1 amplitudes. This finding of selective effects on later rather than earlier occurring components of the LAEP provides objective evidence that some forms of auditory processing are mediated at more central levels of the system. PMID- 15341227 TI - Auditory processing disorders. PMID- 15341228 TI - Communicating health information: an epidemic of the incomprehensible. PMID- 15341229 TI - Portrayal of seasoned nurses. PMID- 15341230 TI - Temporary invasive cardiac pacing. PMID- 15341231 TI - Critical care of patients with obesity. PMID- 15341232 TI - Profiles in dignity: perspectives on nursing and critically ill older adults. PMID- 15341233 TI - Delirium doulas: an innovative approach to enhance care for critically ill older adults. PMID- 15341234 TI - Culturally competent nursing care: a challenge for the 21st century. PMID- 15341235 TI - Amniotic fluid embolism: an obstetric emergency. AB - AFE is an unpredictable, unpreventable, and, for the most part, an untreatable obstetric emergency. Management of this condition includes prompt recognition of the signs and symptoms, aggressive resuscitation efforts, and supportive therapy. Any delays in diagnosis and treatment can result in increased maternal and/or fetal impairment or death. Whereas once the invariable outcome of AFE was death of the mother, today the prognosis is somewhat brighter thanks to increased awareness of the syndrome and advances in intensive care medicine. In any case, intensive care nurses are called on to provide physical, life-saving care to the patient and her fetus. Both during and after the event, supportive care must be administered to the patient's family members, who are dealing with crisis and loss. PMID- 15341236 TI - What is the current status of liver support devices in the United States? PMID- 15341237 TI - Developing an interdisciplinary model of care in a progressive medical care unit. PMID- 15341238 TI - The fourth element of negligence. PMID- 15341239 TI - Mobility protocol: nursing standard of care. PMID- 15341240 TI - Privacy impact assessments--oversight by the Information and Privacy Commissioner. PMID- 15341241 TI - Health privacy in Canada--shedding light on murky waters. PMID- 15341242 TI - The Personal Health Information Protection Act, 2004: Ontario's response to PIPEDA. PMID- 15341243 TI - The application of new privacy legislation in health care. PMID- 15341244 TI - Patterns of cyclooxygenase-2 inhibitor utilization. PMID- 15341245 TI - Utilization patterns of cyclooxygenase-2 inhibitors among patients with arthritis in a managed care setting. AB - When anti-inflammatory/analgesic agents are not well tolerated, patients with arthritis may be prescribed a cyclooxygenase-2 (COX-2) inhibitor. Since these patients often require daily treatment and COX-2 inhibitors are more expensive than nonselective nonsteroidal anti-inflammatory agents, it is important to assess their patterns of use. In this retrospective study, rofecoxib and celecoxib were compared, in a managed care population with arthritis, in terms of average daily medication consumption and cost. Celecoxib was found to be significantly more costly than rofecoxib, and certain factors, such as the treating physician's specialty, correlated with prescribing patterns. Given the high prevalence of arthritic conditions, these results suggest that the selection of a COX-2 inhibitor may substantially affect health care costs. PMID- 15341246 TI - The "truth" in research: setting realistic expectations about outcomes. PMID- 15341247 TI - Generalizing disease management program results: how to get from here to there. AB - For a disease management (DM) program, the ability to generalize results from the intervention group to the population, to other populations, or to other diseases is as important as demonstrating internal validity. This article provides an overview of the threats to external validity of DM programs, and offers methods to improve the capability for generalizing results obtained through the program. The external validity of DM programs must be evaluated even before program selection and implementation are begun with a prospective new client. Any fundamental differences in characteristics between individuals in an established DM program and in a new population/environment may limit the ability to generalize. PMID- 15341248 TI - Interdisciplinary management of chronic disease in primary practice. AB - Primary care physicians are often challenged by the complex needs of patients with chronic disease. Unfortunately, many solo and small group practices lack the interdisciplinary resources essential to chronic disease management. To address this, clinicians and researchers at Rush University Medical Center, Chicago, developed the "Virtual Integrated Practice," a process that creates virtual patient care teams. Preliminary results in four practice sites using it have shown increased referrals to dietitians and improved nutritional compliance but less extensive involvement of social workers and pharmacy team members. PMID- 15341249 TI - [Ultrasound screening and diagnosis of fetal structural abnormalities between 11 14 gestational weeks]. AB - AIM OF THE STUDY: To assess the feasibility of ultrasound screening and diagnosis of structural fetal anomalies at the 11-14 week scan. METHODS: An observational prospective follow up study from March 2000 till May 2003 was performed at three referral centers by seven experienced sonographers with high-resolution ultrasound equipment. 1135 singleton pregnancies between 11+0 and 14+6 weeks gestation (w.g.) participated in the study. The first trimester scan included assessment of fetal number, viability and biometry, nuchal translucency [NT] measurement and fetal anatomy survey performed according to standardized published protocols. Increased NT > or = 95th centile and/or diagnosis of structural fetal anomaly was considered as indication for invasive prenatal diagnosis, early fetal echocardiogram and follow-up scans, including a detailed fetal anomaly scan at 18-22 w.g. and a third scan at 28-32 w.g. Pregnancy outcome was ascertained from hospital records, referring physicians or the patients themselves. RESULTS: The overall prevalence of structural fetal anomalies in the present study was 4.6% (53/1135). 22% (12/53) of the structural anomalies were detected between 11-14 w.g. 9 of those had normal karyotype, and 3 were associated with chromosomal anomalies. Furthermore, 10 cases of increased NT, with or without non-immune hydrops fetalis, were associated with congenital heart disease, rare genetic syndromes and adverse pregnancy outcome later in gestation. The ultrasound detection rate of structural fetal anomalies in the present study increased from 22% (12/53), to 69% (37/53) and 79% (42/53) for the first trimester scan, the first and second trimester scans, and the combination of all three scans, respectively. 21% (11/53) of all structural fetal anomalies were missed by prenatal ultrasound. CONCLUSIONS: The first trimester scan is a method of choice for the diagnosis of major structural fetal anomalies. NT measurement is a useful screening test for chromosomal anomalies. In cases with increased NT subsequent development of congenital heart disease, rare genetic syndromes or adverse pregnancy outcome should be ruled out. At present, the second trimester scan constitutes an indispensable tool for the detection of most structural abnormalities. Even in advanced gestation the prenatal diagnosis of certain anomalies is difficult and often unfeasible. PMID- 15341250 TI - [Significance of fetal factors in prognosis of outcome after labor induction]. AB - Aim of the present mixed prospective and retrospective study is to determine the prognostic significance of fetal factors for the continuity and the final outlet of induction of labor. Factors like fetal weight, placental maturity according Grannum, status of the amniotic membranes and amniotic fluid index by Phelan were examined. Based on the given results the authors conclude that there is no dependence between the expected overweight of the fetus and the continuity and the final outcome of induction of labor. The immature placenta and the increased amniotic fluid index are related with prolonged inductions of labor but the terminal outcome is not influenced by them. Premature rupture of the amniotic membranes cause prolonged inductions and the preserved membranes with an increased frequency of unsuccessful inductions. PMID- 15341251 TI - [Causes of urinary tract infection in pregnant women]. AB - Data of etiology characteristics of uroinfections with pregnant women are presented treated at University Hospital of Obstetrics and Gynecology "Maichin dom" in three periods. Gram negative species of microorganisms (E. coli, proteus, Klebsiella) are the agens of uroinfections in 95.4% of cases in the period 1988/89. After 10-year period gram (-) are the uroinfection in 66.1% of the cases. The growing etiologycal importance of Gram positive species of microorganisms (S. aureus, Enterococcus, GBS) is 33,9% in the period 1997/98 and it is preserved in june 2001/may 2003 too. In cases of long lasting hospitalization or after repeated treatment with antibiotics the probability grows that the agents of uroinfections be polyresistant strains. In the period june 2001/may 2003 in 7% of the cases polyresistant E. coli are isolated, in 2.6% MRSaureus, in 7.6% Ampicillin - R Enterococcus spp. That suggest exact identification of the agen and therapy according to sensitivity to antibiotics. PMID- 15341252 TI - [Can education duning pregnancy improve the perinatal outcome in diabetic pregnancies?]. AB - The aim of the study was to define the role of education for the complex treatment plan of diabetes during pregency and to estimate the impact of education for the reduction of the incidence of maternal and neonatal complications. STUDY DESIGN: A retrospective study of 56 pregnant women with diabetes was carried out. They were hospitalized in the High-risk pregnancy Department from January 1999 till January 2000. All pregnant women were admitted to the hospital in each trimester of pregnancy for control of carbohydrate metabolism and follow-up of pregnancy. All pregnant women were treated by intensified Insulin regime-type basal-prandial. The diet was set according to the needs during pregnancy. 30 pregnant women (group 1) were educated according to the specificity of diabetes during pregnancy. The education was repeated every time the patient was admitted to the hospital. The aim of education was to reach and maintain good metabolic control (HbA1c < 7.5%) in the course of pregnancy. The values of HbA1c before education in group 1 and after final education in group 2 (not educated) were determinated. The incidence of some maternal and perinatal complication-the progress from diabetic nonproliferative retinopathy into proliferative retinopathy, the development of preeclampsia as a causative factor for delivery before 37 g.w, the development of RDS and perinatal deaths were evaluated. RESULTS: There was a significant difference in the mean values of glycosylated Hb. In the group of educated women a good metabolic control was achieved--HbA1c = 7.5%. In the non-educated group the metabolic control was poor- HbA1c = 8.4%, irrespective of the insulin and diet treatment. 25 (84.4%) of the babies born to educated mothers had normal birthweight between 3000 and 4000 grams. 2 (6.6%) of the newborn had birthweight under 3000 gr. and 3 (10%) over 4000 gr. In the group of educated women there was no progress of nonproliferative retinopathy into proliferative type. There was such a progression in 3 women from the noneducated group. There were 22 cases of preeclampsia in both groups. In the group of educated women the preeclampsia was mild, compared to the noneducated group where the form was severe. All women with severe form of preeclampsia (n=8) delivered before 37g. w. There were no perinathal deaths in the educated group, compared to 3 (11.5%) deaths in group 2. 2 babies died antenatally and one after an operative correction for severe congenital heart defect. There were 6 (6.6%) cases of RDS from the educated group compared to 99 (16.6%) from group 2. CONCLUSIONS: The education is a key factor in the complex plan of treatment of pregnant women with diabetes. It creates a positive motivation for the achievement and maintenance of good metabolic control during pregnancy. The incidence of maternal and neonatal complications is higher in the group of non educated women. The education significantly reduces the incidence of maternal and neonatal complications, thus improving the perinatal outcome. PMID- 15341253 TI - [Ferro-Folgamma--a drug for treatment and prophylaxis of iron deficiency anemia in pregnant women]. AB - The problem of iron deficiency anaemia (IDA) in pregnant women is not a new one, but the flashback to it is justified because of the fact that its significance is not appreciated properly. The growing foetus has increased needs of different active for blood formation substances such as iron (Fe), folic acid, vit. B12, which are taken away from the mother. If her supplies are not enough, an anaemic syndrome can be promoted. The only suitable way in the presence of iron deficiency is the administration of drugs for equilibration of iron balance. We used Ferro-Folgamma--a new medicine for our pharmaceutical market, for treatment of IDA in 42 pregnant women and we rendered an account of the results of its administration. All pregnant women had starting data of Hb<110 g/l and Hct<0.33 L/L. One capsule of Ferro-Folgamma contains 100 mg iron sulfate siccatum (that corresponds to 37 mg Fe), folic acid - 5 mg, cyancobalamine 10 mg. It is administered a capsule 3 times daily in 1 to 3 courses of treatment and a course has 20 days duration. After a treatment course was carried out the starting data of Hb were increased with 8.73 g/l, and of Hct with 0.02. In 8 pregnant women after the first therapeutic course, a 20-days - treatment was conducted too and Hb rose with 18.14 g/l in comparison with the starting data, and of Hct - with 0.037. The authors recommend the utilization of Ferro-Folgamma for treatment and prophylaxis of IDA in pregnant women, because of its good gastric acceptance, a few side undesired reactions, and the supplement of Folic acid and vit B12 increases the iron resorbtion. PMID- 15341254 TI - [Comparative study of the endometrial pathology using flexible ambulatory hysteroscopy of patients treated with Tamoxifen]. AB - There were checked out 32 women in concern with genital bleeding receiving Tamoxifen 20 mg daily for a period of an year. The above mentioned were receiving this medicine because of breast cancer with the help of flexible office hysteroscopy. In the first case in group I--patients with Tamoxifen, was found endometrial pathology in 81.25%. In group II--patients without Tamoxifen was found endometrial pathology in 69%. As a result of this investigation the authors confirm that Tamoxifen increase the risk of endometrial pathology. PMID- 15341255 TI - [Combined use of metformin and Diane35 in polycystic ovary syndrome]. AB - The aim of our study was to compare the effect of Metformin applied independently to the effect of Metformin used in combination with Diane35, the latter given only during the first two months of the treatment. METHODS: In this prospective, open clinical study, 30 women with PCOS were included, divided in two groups 15 women each. Group 1 received 850 mg Metformin twice a day and group 2 in which Diane35 was added to the same treatment only during the first two months of the investigation. In addition the following tests have been made--immune reactive insulin, total testosterone, SHBG, free androgen index, anthropometric and some biochemical indices. RESULTS: Much better decreasing of the level of testosterone and free androgen index in group 2, without worsening of the anthropometric and biochemical indices. CONCLUSION: The combination of Metformin with the intermittent application of Diane35 is an appropriate alternative for the pathogenic influence and clinical improvement of the symptoms of androgen excess in cases with PCOS. PMID- 15341256 TI - [Flow cytometric DNA analysis and neoadjuvant chemotherapy of cervix cancer (stage IB2-IIIB)]. AB - The purpose of the present study is to determine the predictive role of the data of flowcytometric DNA-analysis--aneuploidity DNA-index, proliferative index in patients with squamous cervical cancer who was given chemotherapy as a initial treatment. The data of 12 patients in stage IB2-IIIB, who were divided into two groups according to the second-line treatment--surgery of definitive radiotherapy, were analysed. These data were correlated with the respond of the tumor to neoadjuvant chemotherapy (clinical and histopathological) as well as other factors such as stage, size of tumor lesion, grading disease free survival. The method of flowcytometric DNA-analysis was presented briefly. The results we have obtained, although in quiete limited number of patients, are interesting and justify prospective studies of the problem. PMID- 15341257 TI - [Concurent chemoradiotherapy of cervix cancer]. PMID- 15341258 TI - [Trends in the development of the cytological cervical screening]. AB - Cytological method, as a part of cervical screening, demands to keep strictly to the defined standards for its implementation. At the same time the interpretation of the findings appears a subjective assessment of an objective realities. In this connection, the improvement of the screening method becomes the difficult task, which in the last decade resolved at the several directions: preparation of the standard adequate smear (thin preptechnique), automation of the screening and HPV typization as a part of the screening. It is pointed out, that these new technologies reduce the errors, increase the sensitivity and specificity of the diagnostics, as well as show better proportion "cost-effectiveness", comparing to standard screening. PMID- 15341259 TI - [Mola hydatidosa of the uterine tube]. AB - A case of only rare localization of hydatidiform mole in fallopian tube is described. This localization is, recent clinical as ectopic pregnancy. In the last 25 years (1974-1999) was described only 30 cases of tubal localysation of hydatidiform mole. PMID- 15341260 TI - [Difficulties and biases of interpretation in cases of the trophoblast disease]. AB - The trophoblastic disease (TD) is an unifying term for the nosologicunits: Mola hydatidosa, Mola hydatidosa invasivum, Chorionepithelioma. This kind of trophoblast pathology has geographic differences in the expression but the its overall incidence is not high. The clinicians often fail to consider the possibility of trophoblastic disease due to its low incidence rate. The forms of TD have clinical manifestations that are not specific. There are principles, which taken into account, could help the clinicians put the right diagnosis. Two cases of the clinical practice are reviewed in maintenance of this opinion. PMID- 15341261 TI - [Perforation of duodenal ulcer in the puerperium--a case report]. AB - The authors present a case of perforated duodenal ulcer in the puerperium at a patient who underwent Ceaserian section. Urgently 12 hours after the perforation relaparotomy was made. The exit of the illness depends on the precise and early diagnosis and subsequent surgical treatment. PMID- 15341262 TI - [Contemporary diagnostic criteria for polycystic ovary syndrome]. AB - Authors review the diagnostic criteria for polycystic ovary syndrome according to the up to date knowledge and understandings. The most usual pitfalls and the diversity of the syndrome are discussed. PMID- 15341263 TI - Hormone substitution therapy in women operated for adenomiosis and endometriosis. PMID- 15341264 TI - [Behavioral genetics and neurogenetics and their development in Russia]. AB - The problems of behavioral genetics and neurogenetics and development of these fields in Russia are considered. PMID- 15341265 TI - [Changes in the expression of monoaminergic genes under the influence of repeated experience of agonistic interactions: from behavior to gene]. AB - The role of genetic and environmental factors as well as brain neurochemistry in regulating aggressive and submissive behaviors in animals are considered. We present a review of data on changes in brain monoaminergic activity (synthesis, catabolism, receptors) and on the expression of monoaminergetic genes under repeated daily agonistic confrontations in male mice. A repeated experience of aggression was shown to result in the total activation of the dopaminergic system and the inhibition of the serotonergic one. This was accompanied by a decrease in the mRNA level of the catechol-O-methyltransferase gene in the midbrain and an increase of the mRNA level of the dopamine transporter and tyrosine hydroxylase genes in the ventral tegmental area of aggressive male mice. Repeated experience of social defeats produced dynamic changes in the serotonergic system of some brain areas and an increase of the mRNA level of the serotonin transporter and monoamine oxidase A genes in the midbrain raphe nuclei. Theoretical and methodological possibilities of the proposed ethological approach for studying molecular mechanisms of agonistic behavior are discussed in the context of the fundamental problem of investigating the ways of regulation from behavior to gene. PMID- 15341266 TI - [Architecture of the X chromosome, expression of LIM kinase 1, and recombination in the agnostic mutants of Drosophila: a model of human Williams syndrome]. AB - As the Human Genome and Drosophila Genome Projects were completed, it became clear that functions of human disease-associated genes may be elucidated by studying the phenotypic expression of mutations affecting their structural or functional homologs in Drosophila. Genomic diseases were identified as a new class of human disorders. Their cause is recombination, which takes place at gene flanking duplicons to generate chromosome aberrations such as deletions, duplications, inversions, and translocations. The resulting imbalance of the dosage of developmentally important genes arises at a frequency of 10(-3) (higher than the mutation rate of individual genes) and leads to syndromes with multiple manifestations, including cognitive defects. Genomic DNA fragments were cloned from the Drosophila melanogaster agnostic locus, whose mutations impair learning ability and memory. As a result, the locus was exactly localized in X-chromosome region 11A containing the LIM kinase 1 (LIMK1) gene (CG1848), which is conserved among many species. Hemizygosity for the LIMK1 gene, which is caused by recombination at neighboring extended repeats, underlies cognitive disorders in human Williams syndrome. LIMK1 is a component of the integrin signaling cascade, which regulates the functions of the actin cytoskeleton, synaptogenesis, and morphogenesis in the developing brain. Immunofluorescence analysis revealed LIMK1 in all subdomains of the central complex and the visual system of Drosophila melanogaster. Like in the human genome, the D. melanogaster region is flanked by numerous repeats, which were detected by molecular genetic methods and analysis of ectopic chromosome pairing. The repeats determined a higher rate of spontaneous and induced recombination. including unequal crossing over, in the agnostic gene region. Hence, the agnostic locus was considered as the first D. melanogaster model suitable for studying the genetic defect associated with Williams syndrome in human. PMID- 15341267 TI - [The role of brain serotonin in the expression of genetically determined defensive behavior]. AB - The review summarizes the results of long-term studies on the role of the brain mediator serotonin and genetic predisposition to various types of defensive behavior. The involvement of the serotonergic brain system in the mechanisms of genetic control of both active and passive defensive responses has been established using silver foxes, Norway rats of S40 selection for low and high aggressiveness to humans, aggressive mice with genetic knockout of monoaminoxidase A, and S40 rats selected for predisposition to passive defensive response of freezing (catalepsy). The changes in the serotonergic 5-HT1A-brain receptors of rats genetically predisposed to different strategies of defensive behavior were similar. However, the activity of the key enzyme of serotonin biosynthesis and the brain structures, in which serotonin metabolism was altered, significantly differed with regard to the preferred strategy. The conclusion was drawn that the 5-HT1A-receptors and enzymes of serotonin metabolism in the brain are involved in implementing genetic control of defensive behavior. Expression of the 5-HT1A-brain receptors was suggested to determine the levels of fear and anxiety and, consequently, the predisposition to defensive behavior, whereas the preferred strategy of defensive response (active or passive defensive) depends on genetically determined features of serotonin metabolism in the brain structures. PMID- 15341268 TI - [Hereditary catalepsy: genetic and molecular mechanisms of catalepsy in mice]. AB - The results of experiments on the inheritance and neurobiological mechanism of high predisposition to tonic immobility (catalepsy) in CBA mice are discussed. Genetic analysis has demonstrated a monogenic inheritance of the predisposition to catalepsy. A set of polymorphic microsatellite markers has been used to demonstrate that the predisposition to catalepsy is linked to the distal fragment of mouse chromosome 13, which contains the gene of the 5-HT1A-serotonin receptor. Pharmacological and biochemical evidence for the association between hereditary catalepsy and 5-HT1A-receptor dysfunction are presented. The use of CBA mice for studying the mechanisms of depression and the effects of antidepressants is discussed. PMID- 15341269 TI - [Stem cells in neurogenetics]. AB - Genetically controlled proliferation and differentiation of stem nerve cells have been demonstrated to be among the repair mechanisms of the nervous system. Another such process is differentiation of neuroblasts from the cambial reserve. PMID- 15341270 TI - [An experiment on fox domestication and debatable issues of evolution of the dog]. AB - This paper is a review of the results of the authors obtained in a long-term experiment on fox domestication. Debatable issues of dog evolution are discussed in light of these results. It is demonstrated that genetic physiological mechanisms of the behavior transformation during selection and the nature of the arising phenotypic changes are associated with retarded development of corresponding ontogenetic processes. As a result of this retardation, the adult animals retain juvenile traits of behavior and morphology (the phenomenon of neoteny). The role of hormonal changes caused by domestication in the evolutionary origin of neoteny is discussed. PMID- 15341271 TI - [Lorenz was right, or does aggressive energy accumulate?]. AB - Evidence supporting the fact that inherited mechanisms of regulation of aggressive behavior as a result of a repeated experience of aggression ending in victories are transformed into pathological mechanisms based on accumulation of neurochemical shifts in the brain, enhancing aggressiveness, and forming aggressive motivation in aggressive winners. This confirms the concept by Lorenz on the existence of a mechanism (but not instinct) of a spontaneous accumulation of aggressive energy that needs a discharge and formation of permanent attraction to manifestation of aggression. PMID- 15341272 TI - [Molecular genetic analysis of hereditary neurodegenerative diseases]. AB - The review summarizes the results of a decade of molecular genetic studies of several high-incidence hereditary neurodegenerative diseases, including primary parkinsonism, various forms of hereditary dystonia and ataxia, polyglutamine disorders, hepatolenticular degeneration, essential tremor, etc. Various relevant mutations were studied. The character and frequencies of particular mutations and the corresponding genetic disorders were established for the Russian population. Particular genotypes were associated with various clinical variants of the diseases. Genetic loci were identified for several unique hereditary diseases of the nervous system (X-linked cerebellar hypoplasia, an atypical form of autosomal recessive muscular dystrophy, etc.). Nosological positions of the relevant clinical forms were clarified on the basis of the molecular genetic data. Protocols were developed for direct or indirect DNA diagnostics of the diseases under study to improve medical genetic counseling and prevention of new disease cases in affected families. PMID- 15341273 TI - [Catatonia or depression: the GC rat strain as an animal model of psychopathology]. AB - The utility of "incomplete" genetic animal models of human diseases, in particular, psychoses, is discussed. The GC rat strain selected for predisposition to cataleptic reactions is described. It is shown that in many of their characteristics, GC rats are similar to schizophrenic and depressive patients. A possibility that akinetic catatonic states and depressions, hyperkinetic catatonic states and mania share common mechanisms is discussed. It is hypothesized that the GC strain may be an incomplete model of the common genetic and pathogenetic core of schizophrenic substuporous states and depression, which suggests the importance of returning to the issue of a unitary psychosis (Einheitpsychosis). PMID- 15341274 TI - [Analysis of the DRD4 gene polymorphism in populations of Russia and neighboring countries]. AB - Allele and genotype frequencies of the VNTR polymorphism in the third exon of human DRD4 gene were determined in 544 individuals living in Russia (Russians, Bashkirs, Tatars, and Mordovians) and in the neighboring countries (Kazakhs and Ukrainians). The data obtained were compared with the allele frequency distribution patterns reported for the populations of Eurasia. Similarly to other Eurasian populations, in our population samples R4 allele was prevalent (64 to 87%). The frequency of this allele in the populations of Western Europe constitute 61 to 71%, while in the populations of Asia it varies from 74 to 96%. In this respect, the populations studied occupied the intermediate position. In the samples examined the R7 allele frequency decreased from 7% in Ukrainians to 1% in Bashkirs, while in Kazakhs and Mordovians the allele was absent. This finding was consistent with the R7 allele distribution pattern in the populations of Eurasia, characterized by higher frequency in the West and lower frequency or absence of the allele in the East. In the group of 22 Eurasian populations, the R7 allele frequency negatively correlated with the frequency of the R4 allele (r = -0.86 at P < 0.001). Unlike the R4 and R7 alleles, the frequency of which changed in the eastward direction, the R2 allele frequency distribution displayed slightly expressed latitudinal increase southwards. The DRD4 genotype distribution deviated from the equilibrium in most of the samples examined. In some samples, statistically significant increase of the R2/R2 homozygotes frequency was demonstrated. One of the possible explanations of this phenomenon is assortative mating with respect to phenotypic (behavioral) allele manifestation. The data obtained can serve as the basis for the investigation of the possible role of the DRD4 alleles as the risk factors for the development of alcoholism and other types of addictions. PMID- 15341275 TI - [Polymorphism of the serotonin system genes in some Finno-Ugric populations]. AB - Polymorphic sites in the genes encoding monoamine oxidase A (MAO-A), serotonin transporter (hSERT) and 5-HT2A receptor were typed in Khant and Komi ethnic groups with the purpose of revealing possible interpopulation differences in genotype and allele frequencies. No statistically significant differences in the hSERT and 5-HT2A gene frequencies were detected. At the same time, the populations examined had statistically significantly different MAO-A genotype and allele frequencies. These results obtained indicate the prevalence of the site gain alleles of the EcoRV and Fnu4HI RFLP loci at the MAO-A gene in Komis and the of the corresponding site loss alleles in Khants. PMID- 15341276 TI - [The interstrain differences in the effects of D-amphetamine and raclopride on dorsal striatum dopaminergic system in KM and Wistar rats (microdialysis study)]. AB - The levels of dopamine (DA) was determined by intracerebral microdialysis in vivo in KM rats selected for high audiogenic epilepsy, and in Wistar rats selected for nonsusceptibility to loud sound. The basal level of dopamine was 25% higher in the KM rats (P < 0.05). A single amphetamine injection (1 mg/kg body weight, intraperitoneously) caused a significant increase in the DA basal level up to 250 260% in animals of both genotypes. However, in Wistar rats, the level of DA reached maximum as soon as 20 min after amphetamine administration, whereas in KM rats, this happened only after 120 min. After a single injection of the antagonist of D2 and D3 dopamine receptors raclopride (1.2 mg/kg of body weight, intraperitoneously), an increase in the level of DA was similar in amplitude in rats of both genotypes (up to about 210%); however, this occurred 20-30 and 100 min after raclopride administration to Wistar and KM rats, respectively. This evidence suggests that the genetic defect of KM rats, namely, the high level of audiogenic epilepsy, is caused by abnormalities of the neuromediator brain systems and presumably accompanied by the regulatory gene dysfunction. PMID- 15341277 TI - [Analysis of the glutathione S-transferase P1 gene Ile105Val polymorphism in the patients with sporadic motor neuron disease from Russia]. AB - Ile105Val polymorphism in exon 5 of glutathione S-transferase (GSTP1) gene was examined in a group of patients with motor neuron disease (MND) and control sample. No statistically significant differences in the allele and genotype frequency distributions between the samples examined were demonstrated. We conclude that Ile105Val polymorphism is not associated with the risk of the disease development in the patients from Russia with sporadic form of MND. PMID- 15341278 TI - [Cloning and analysis of a new neurogene in the mouse]. AB - The previously described gene sip1 belongs to transcription factors of the zinc finger family. It has been ascertained recently that this gene is involved in TGF signaling cascade. Mutations in human gene sip1 cause Hirschprung syndrome. The expression of gene sip1 during embryonic mouse development was studied by in situ hybridization and immunostaining. Starting at E12.5, sip1 transcripts are present in a number of tissues: in the cortical plate, ventricular zone of the basal ganglion, thalamus, pons and midbrain, in specific nuclei of the brain stem and in the dorsal part of the spinal cord. In the developing cerebral cortex, sip1 expression is region-specific. In the brain of adult mice, sip1 expression is mostly detected in hippocampus, dentate gyrus, and white matter of the neocortex. Sip1 protein expression in the cerebral cortex is mostly confined to glutamatergic neurons. PMID- 15341279 TI - Volunteerism. PMID- 15341280 TI - Making a difference. PMID- 15341281 TI - Applause for compassionate efforts. PMID- 15341282 TI - National Children's Dental Health Day: Give Kids A Smile. PMID- 15341283 TI - Expanded dental care for the underserved of Hamakua and North Hawaii. PMID- 15341284 TI - The Queen's Medical Center Dental Clinic: committed to making a significant difference. PMID- 15341285 TI - Special Olympics, Special Smiles. PMID- 15341286 TI - [Expert evaluation of medical care quality in otorhinolaryngology]. AB - Defects related with rendering medical care (DMC) in the managerial and information-deontological spheres of healthcare are evaluated. Reasons of the above defects and the related peculiarities are elucidated; the necessity in eliminating them is substantiated. The authors put forward their own definition of DMC on the basis of an independent analysis. PMID- 15341287 TI - [An algorithm of forensic-histological examination]. AB - A detailed algorithm of forensic-histological examinations supplementing "The Instruction on organization and execution of examinations by Bureau of forensic medical expertise" is suggested in the paper. The algorithm envisages examination of the preparation in flow light without microscope, identification of an organ and/or tissue as well as an evaluation of the integrity of histo-architectonics and of the pronouncement degree of cadaveric changes. A regulated description of the preparation is recommended by using the generally accepted terminology; peculiarities related with examination of different organs are elucidated. Features, which are most informative in solving the forensic-medical issues, are specified. Methodological approaches to formulating the forensic-histological diagnosis are defined with respect to diagnosis specificity. PMID- 15341288 TI - [Mathematic modeling of morphogenesis of the liver and spleen in fetus and newborns for the purpose of determining the gestation age]. AB - Fragments of the liver and spleen obtained from 140 cadavers of fetus and newborns with a gestation age (GA) ranging from 21 to 41 weeks were morphometrically investigated. The obtained data were statistically processed. A set of equations was designed on the basis of the multi-factor regression analysis for the GA determination. The study results are recommended for practical use in the forensic-medical expertise in cases of perinatal death. PMID- 15341289 TI - [A study of cadaveric mycobiota at the late stages of post-mortem changes within the forensic-medical verification of death coming and of bury place]. AB - Data are described on the late stages of fungus populating in cadaveric remains (soil and other fungi). By week 2 after bury, the soil fungi grow through the cloths and skin of cadaver with the species mycobiotic composition of remains corresponding, on the whole, to a bury place. The woodland soils are distinguished through a more diversified species composition and total mycobiotic content, which can be valuable in establishing the fact and route of transfers of cadaver or of its remains even at late cadaveric changes. PMID- 15341290 TI - [Determination of the severity of endogenous intoxication at the level of average molecular peptides]. AB - A possibility was demonstrated of using the method related with determination of the level of average-weight molecules or average molecules (AM) for the diagnosis of sudden death in forensic medical expertise of cadaver. Blood plasma was investigated according to the method of M.I. Gabriekyan modified by Yu. V. Pervushin, which involves deproteinization of samples with subsequent spectrophotometry at 254 and 280 nm. The AM levels were studied in different pathologies including cardiovascular diseases, alcoholic and narcotic intoxications and pneumonia, caused by a variety of agents, as well as the syndrome of sudden children's death. An analysis of the obtained results denotes a possibility of using the postmortem determination of the AM level concurrently with determination of an endogenous intoxication degree in the forensic medical practice for establishing the cause and mechanisms of nonviolent death. PMID- 15341291 TI - [Potentialities of histological examinations of the spleen in the diagnosis of alcoholic and narcotic intoxication]. AB - Qualitative changes observed in the spleen in chronic narcotic and alcoholic intoxication are described. The below signs are recommended for use in the diagnosis of chronic exogenous intoxication: an increased thickness and sclerosis of the splenic connective structures and myelosis of the red pulp. The diagnostic criteria of chronic intoxication with intravenous opiates are the below changes in the spleen: hyperplasia of the splenic follicles, germinative foci in the follicles' stases and folds in vessels, edema of the vascular wall, hyperemia of follicular arteries and diapedetic hemorrhages. Hypoplasia of the follicles and venous hyperemia of the spleen should be regarded as signs of chronic alcoholic intoxication. The data obtained within the case study can be used by forensic medical experts in the diagnosis of narcotic and alcoholic intoxication. PMID- 15341292 TI - [Ethanol-oxidizing enzymes of the adrenal glands in acute alcoholic intoxication]. AB - Complex morphological-and-histological examinations of the adrenal glands were performed on actual forensic-medical samples in acute alcoholic intoxication (AAI), i.e. 107 death cases of men and women, aged 17 to 60, due to mechanical trauma, lethal ethanol intoxication and somatic complications of chronic alcoholism. A differentiated distribution of ethanol-oxidizing enzymes was defined, i.e. of alcohol-dehydrogenase and acetyl-dehydrogenase in the structural and-functional zones of the adrenal cortex and medullary substance. A differential activity of enzymes was established that correlated with the level of exogenous alcoholemia. Sets of morphological signs related with examinations of the adrenal glands, which enabled a differential diagnosis of death causes in AAI, were specified. PMID- 15341293 TI - [Determination of some somatic body dimensions by osteometric foot measurements]. AB - Methods of diagnosis of somatic life-time peculiarities of man (body height, shoulders breadth and dimensions of extremities and their segments) were elaborated on the basis of a study of interrelations between the human body-built features, on the one hand, and measurement characteristics of the foot, on the other hand, by using the modern programs of multi-measurement statistics. Isolated skeleton bones of osseous complexes and fragments of native and burnt bones were the research objects. Footwear and cloths items, which belonged to an actual missing person, can be identification objects. The paper is primarily addressed to forensic medical experts and can be useful for criminalists, anthropologists and archeologists. PMID- 15341294 TI - [A definition of the nomenclature of allele variants and a description of allele polymorphism within a molecular-genetic individualization system based on the pentanucleotide tandem replication of HUMCD4]. AB - While analyzing the available published data, we found significant differences in definition of alleles of the HUMCD4 polymorphic chromosome locus. It is an obstacle for comparing the expertise results obtained while using the locus as molecular individualization system in different laboratories and, as a consequence, it hinders the use of the said marker in building up a reference database. The structure of the HUMCD4 locus was analytically investigated and the distribution of the locus alleles was systemized in a sample of 407 persons (citizens of Russia who are not blood relatives) within the present case study for the purpose of a detailed definition of its allele's characteristics. We worked out recommendations related with the nomenclature standardization of alleles of the HIMCD4 locus and specified the main population features of the locus polymorphism on the bases of the studied sample of Russia's citizens. The results can be used as reference parameters in interpreting the findings of the molecular-genetic identification expertise and examinations and in building up databases. PMID- 15341295 TI - [The role of carrier-object in investigations of cigar-butts]. AB - A comprehensive series of experiments was undertaken to find the reasons why the cigar-butt stuff, removed from accident places for investigation at forensic biological laboratories, affects different sera. A number of such reasons were established. Methods are suggested to eliminate the cigar-butt impact with the aim of ensuring the most accurate results while determining the group adherence of saliva on the cigar-butts. PMID- 15341296 TI - [Detection of A and B antigens of the ABO system in a single-hair stem]. AB - A method was elaborated for detection of the system ABO antigens in a single-hair stem, containing no hair bulb or vaginal tunics, through using concurrently electrophoresis in the agarous or agar gel as well as through extracting antigens from a certain gel area onto the gauze with performing subsequently the absorption-elution reaction. PMID- 15341297 TI - [The material-and-technical equipment of state forensic medical bureaus of Russia's defense ministry: realities and outlooks]. AB - Issues related with the material-and-technical outfit of state forensic medical bureaus of Russia's defense ministry are under discussion. The present-day condition of their instrumentation degree is described; methods are suggested of how to build up the material-and-technical base by using the limiting form system with the "conclusive medicine" principles. PMID- 15341298 TI - [Chemical-and-toxicological evaluation of results of determining the opiates (heroin, morphine and codeine) in urine within the forensic medical expert practice]. PMID- 15341299 TI - America's health care system in crisis: the case for telemedicine. PMID- 15341300 TI - Preemptive measures: a guide to potential pitfalls in a new telehealth program. PMID- 15341301 TI - Telemedicine then and now: the legacy of Dorothy McHugh. Special 2004 home care & hospice technology update. PMID- 15341302 TI - Rep. Max Burns: a political leader makes home care history. PMID- 15341303 TI - Telemedicine revolution makes the home the center of health care. PMID- 15341304 TI - Japan seeks robotic help in caring for the aged. PMID- 15341305 TI - Achieving heroic customer service. PMID- 15341306 TI - Marketing & customer service in a digital world. PMID- 15341307 TI - Intimate moments. PMID- 15341308 TI - Introducing the new "high tech, high touch" landscape. PMID- 15341309 TI - The story of 2,4,5-T: a case study of science and societal concerns. PMID- 15341310 TI - Long overlooked historical information on Agent Orange and TCDD following massive applications of 2,4,5-T-containing herbicides, Eglin Air Force Base, Florida. AB - BACKGROUND: From 1961-1971, The Air Development Test Center, Eglin Air Force Base (AFB), Florida, developed, tested, and calibrated the aerial spray systems used in support of Operation RANCH HAND and the US Army Chemical Corps in Vietnam. Twenty major test and evaluation projects of aerial spray equipment were conducted on four fully instrumented test grids, each uniquely arrayed to match the needs of fixed-wing, helicopter, or jet aircraft. Each of the grids was established within the boundary of Test Area 52A of the Eglin Reservation. METHODS: The tests, conducted under climatic and environmental conditions similar to those in Vietnam, included the use of the military herbicides (Agents) Orange, Purple, White, and Blue. Approximately 75,000 kg of 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and 76,000 kg of 2,4-dichlorophenoxyacetic acid (2,4-D) were aerially disseminated on an area of less than 3 km2 during the period 1962-1970. Data from the analysis of archived samples suggested that an estimated 3.1 kg of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), present as a contaminant, were aerially released in the test area. Because most of the vegetation had been removed before establishing the test site in 1961, there was an opportunity to follow ground-based residues independent of canopy interception, and the resulting high solar exposure of initial residues. Studies of the soils, fauna, flora, and aquatic ecosystems of the test grids and associated perimeters of Test Area C-52A (an area totally more than 8 km2) were initiated in 1969 and concluded in 1984. RESULTS AND DISCUSSION: Data from soil samples collected from 1974 through 1984 suggested that less than one percent of the TCDD that was present in soil when sampling began persisted through the ten-year period of sampling. More than 340 species of organisms were observed and identified within the test area. More than 300 biological samples were analyzed for TCDD and detectable residues were found in 16 of 45 species examined. Examination of the ecological niches of the species containing TCDD residues suggested each was in close contact with contaminated soil. Indepth field studies, including anatomical, histological and ultrastructural examinations, spanning more than 50 generations of the Beachmouse, Peromyscus polionotus, demonstrated that continual exposure to soil concentrations of 0.1 to 1.5 parts-per-billion (ng/g) of TCDD, had minimal effects upon the health and reproduction of this species. CONCLUSIONS: Since Agent Orange with its associated TCDD contaminant was aerially disseminated on the test grids, Test Area C-52A provided a 'field laboratory' for what may have happened in Vietnam, had there been no intercepting forest cover. However, in Vietnam a 'typical' mission would have disseminated 14.8 kg of 2,4,5-T/ha, most of which was intercepted by the forest canopy, versus the 876 kg 2,4,5-T/ha on the test grid at Eglin. Moreover, each hectare on the Eglin test grid received at least 1,300 times more TCDD than a hectare sprayed with Agent Orange in Vietnam. The disappearance or persistence of TCDD is dependent upon how it enters the ecosystem. Spray equipment test and evaluations missions at Eglin were generally scheduled and conducted with environmental conditions that were optimal for spray operations. This suggests that conditions favorable for dissemination of herbicide were the same conditions favorable for photodegradation of TCDD. It was likely that 99 percent of the TCDD never persisted beyond the day of application. No long-term adverse ecological effects were documented in these studies despite the massive quantities of herbicides and TCDD that were applied to the site. Reviews by the US Environmental Protection Agency and the National Academy of Sciences' Institute of Medicine did not address the fate of Agent Orange and TCDD as described in these studies from Eglin AFB, Florida. PMID- 15341311 TI - Sensitized photooxygenation of the fungicide furalaxyl. AB - BACKGROUND: The photolysis of pesticides is of high current interest since light is one of the most important abiotic factors which are responsible for the environmental fate of these substances and may induce their conversion into noxious products. The action of light can also be mediated by oxygen and synthetic or naturally occurring substances which act as sensitizers. Our objective in this study was to investigate the photochemical behaviour of the systemic fungicide furalaxyl in the presence of oxygen and various sensitizers, and to compare the toxicity of the main photoproduct(s) to that of the parent compound. Previous reports on the direct photolysis of the pesticide demonstrated a very slow degradation and the only identified photoproducts were N-2,6-xylyl D,L-alaninare and 2,6-dimethylaniline. METHODS: Solutions of furalaxyl in CH3CN were photooxygenate using a 500W high-pressure mercury lamp (through a Pyrex glass filter, lambda>300 nm) or a 650W halogen lamp or sunlight and the proper sensitizer. When sunlight was used, aqueous solutions were employed. The photodegradation was checked by NMR and/or GC-MS. The photoproducts were spectroscopically evidenced and, when possible, isolated chromatographically. Acute toxicity tests were performed on the rotifer Brachionus calyciflorus, the crustacean cladoceran Daphnia magna and the anostracan Thamnocephalus platyurus, while chronic toxicity tests (sublethal endpoints) comprised a producer, the alga Pseudokirchneriella subcapitata and the crustacean Ceriodaphnia dubia, as a consumer. RESULTS AND DISCUSSION: In the presence of both oxygen and sensitizer, furalaxyl underwent rapid photochemical transformations mainly to N-disubstituted formamide, maleic anhydride and a 2(5H)-furanone derivative. The formation of these products was rationalized in terms of a furan endoperoxide intermediate derived from the reaction of furalaxyl with active dioxygenated species (singlet oxygen, superoxide anion or ground state oxygen). The 2(5H)-furanone exhibited a higher toxicity than the parent compound. CONCLUSION: This work reports the first data on the photosensitized oxygenation of furalaxyl with evidence of the high tendency of the pesticide to undergo photodegradation under these conditions leading, among other things, to a 2(5H)-furanone, which is more toxic than the starting furalaxyl towards aquatic organisms. RECOMMENDATIONS AND OUTLOOK: Investigation highlights that the photolytic fate of a pesticide, although quite stable to direct photoreaction due to its low absorption of solar radiation at ground level, can be significantly influenced in the environment by the presence of substances with energy or electron-transfer properties as natural dyes, e.g. chlorophyll, or synthetic pollutants, e.g. polycyclic aromatic hydrocarbons (PAH). PMID- 15341312 TI - Land-based sources of marine pollution: organochlorine pesticides in stream systems. AB - BACKGROUND: Organochlorine pesticides (OCPs) have been dispersed ubiquitously in the environment. Bottom sediments act as sinks for these compounds and their concentrations often reflect the degree of anthropogenic pollution. This study was designed to evaluate the occurrence and distribution of OCPs in superficial streambed sediments and their relation to land use in two creeks that contribute to the coastal pollution of the southeastern region of Argentina. METHODS: Sampling sites were selected by a combination of land use and stream type. Las Brusquitas creek, which passes through vast agricultural areas, and La Tapera creek which originates in a natural wetland and passes through horticultural farming and urban areas. OCPs quantification was carried out by GC-ECD. RESULTS AND DISCUSSION: Results showed similar total OCP concentrations in sediments from both creeks in the range of 6-25 ng/g dry wt. However, when OCPs were expressed in ng/g total organic carbon (TOC), La Tapera creek presented 4-fold higher total levels as a consequence of a higher OCP input during the recent past in that watershed. La Tapera outfall showed 4-fold higher levels than that seen in Las Brusquitas, although both values were below the sediment quality criteria demanded to protect wildlife. Sigmaendosulfans, sigmaDDTs and sigmachlordanes were the main OCP group in all samples, with Endosulfan sulfate being the most frequent and abundant compound. The predominance of metabolites with respect to parent compounds suggests a contamination mainly by runoff from aged and weathered agricultural soils. CONCLUSIONS: Despite OCPs being banned, they still exist in creek sediments from the studied region, representing continuous contributions of land-based source contaminants to the marine environment. OUTLOOK: Future research on OCP levels in suspended sediments is recommended in order to determine the total OCP concentrations in the selected stream systems. PMID- 15341313 TI - Assessing the toxicity and teratogenicity of pond water in north-central Minnesota to amphibians. AB - BACKGROUND: Incidence of amphibian deformities have increased in recent years, especially in the northern region of the United States. While many factors have been proposed as being responsible for generating deformities (e.g., contaminants, ultraviolet radiation [UV], parasites), no single cause has been definitively established. METHODS: To determine whether waterborne chemicals are responsible for amphibian deformities in ponds in north-central Minnesota, we deployed semipermeable membrane devices (SPMDs) in an impacted and a reference site to accumulate lipophilic contaminants. We then exposed native tadpoles (northern leopard frogs; Rana pipiens) to the SPMD extracts combined with two agricultural pesticides (atrazine, carbaryl) at two levels of UV radiation. RESULTS AND DISCUSSION: UV radiation alone caused a slight increase in hatching success and tadpole growth rate. Deformity rate among hatchlings was high following exposure to SPMD extracts from the reference site in the absence of UV, suggesting that chemicals present at this site are broken down by UV to less harmful forms, or become less bioavailable. Conversely, impacted site SPMD extracts caused hatchling deformities only in the presence of UV, suggesting that UV potentiates the teratogenicity of the compounds present there. Impacted site SPMD extracts significantly increased the number of bony triangles among metamorphs, a common deformity observed at this site. The incidence of skin webbings increased significantly with SPMD extracts from both sites as well as with our pesticide control containing atrazine and carbaryl alone. CONCLUSIONS: Higher deformity rates among tadpoles reared in the presence of UV radiation and SPMD extracts from sites where deformities are common indicates a chemical compound (or compounds) in the water at this site may be causing the deformities. RECOMMENDATIONS AND OUTLOOK: It is important to examine the effects of chemical stressors in the presence of other natural stressors (e.g., UV radiation) to gain a better understanding of how multiple stressors work to impact amphibians and amphibian populations. PMID- 15341314 TI - The identification of readily bioavailable pollutants in Lake Shkodra/Skadar using semipermeable membrane devices (SPMDs), bioassays and chemical analysis. AB - GOAL, SCOPE AND BACKGROUND: Lake Shkodra/Skadar is the largest lake in the Balkans region and located on the border between Albania to the south and Montenegro to the north. Because of the wide range of endemic, rare or endangered plant and animal species it supports, Lake Shkodra/Skadar and its extensive associated wetlands are internationally recognised as a site of significance and importance (Ramsar site). In recent years, social and economic changes in both Albania and Montenegro have lead to unprecedented levels of urban and industrial effluent entering the lake. Of particular concern is the increasing input of toxic hydrophobic organic pollutants (HOPs) into the lake and the degree to which these compounds are available for uptake by aquatic biota. Semipermeable membrane devices (SPMDs) have been shown to sample the readily bioavailable fraction (dissolved phase) of waterborne HOPs and in doing so provide relevant data for exposure assessment. The aim of the current study was to use SPMD-based sampling in conjunction with appropriate bioassays and chemical analysis to identify readily bioavailable HOPs in the lake. METHODS: SPMDs were constructed and deployed at three sites in the Albanian sector and three sites in the Montenegrin sector of Lake Skadar/Shkodra for 21 days. Following the dialytic recovery of target analytes and size exclusion chromatographic clean-up, aliquots of SPMD samples were subjected to GC-MS scan analysis for major components, GC-MS SIM analysis for 16 priority pollutant polycyclic aromatic hydrocarbons (PP-PAHs) and assayed for EROD-inducing, estrogenic and mutagenic potential using rainbow trout liver cells (RTL-W1), the yeast estrogen screen (YES) and the Ames Test, respectively. RESULTS AND DISCUSSION: A total of 39 compounds were tentatively identified in SPMD samples from the six sampling sites. Alkylated PAHs were the most abundant and ubiquitous compounds present along with various sterols and sterol derivatives. Numerous other compounds remain unidentified. 15 of the 16 targeted PP-PAHs were present in samples from one or more of the sampling sites indicating these compounds are both readily bioavailable and widely distributed in Lake Shkodra/Skadar. Total PP-PAH concentrations ranged between 3991 ng/SPMD and 10695 ng/SPMD. Bioassays carried out on SPMD samples revealed significant EROD-inducing and estrogenic potential at five of the six sampling sites indicating toxicologically relevant compounds are readily available for uptake by resident aquatic biota. EROD-inducing potential was positively correlated with targeted PP-PAH concentration (r2 = 0.74). However, comparison of bioassay- and analytically-derived toxicity equivalents revealed targeted PP-PAHs were responsible for less than 0.06% of the total EROD-inducing potential. CONCLUSIONS AND OUTLOOK: The combination of SPMD-based sampling with appropriate bioassays and chemical analysis provided an effective tool for the identification of environmentally relevant waterborne pollutants in Lake Shkodra/Skadar. Our results show that toxicologically relevant HOPs including EROD-inducing and potentially estrogenic compounds are widely distributed in the lake and readily available for uptake by aquatic biota. Our results also suggest that alkylated PAHs rather than parent compounds may be of greater toxicological relevance in the lake. As anthropogenic influences continue to increase, SPMD-based sampling is expected to play a central role in future research concerned with the identification, monitoring and assessment of the risk posed by HOPs to Lake Shkodra/Skadar's aquatic biota. PMID- 15341315 TI - Fe0-based system as innovative technology for degrading trichloromethane: redox removal characteristics. AB - BACKGROUND: The spent waste of aliphatic chlorinated solvents has caused severe deterioration of groundwater quality. Trichloromethane (TCM), which shows health and toxicological effects on human beings, was selected as a model compound to be dechlorinated through a redox system. METHODS: The Fe0-based system including Fe0/H2O, Fe0/UV, Fe0/H2O2, and Fe0/UV/H2O2 was explored to evaluate its performance in dechlorinating TCM. H2O2 was dosed at later reaction time points to initiate Fenton or photo-Fenton reactions. The first two systems demonstrate the reductive dechlorination of TCM by Fe0-released electrons, while the latter two show dechlorination of TCM by both electron reduction and hydroxyl radical oxidation. The system parameters of TCM remaining, Cl- buildup, Fe2+ accumulation, H2O2 residue, and ORP were measured to describe different redox characteristics of TCM dechlorination. The Cl- buildup was used as a way to describe the degree of TCM dechlorination in an open reaction system. RESULTS: Reductive dechlorination efficiencies of TCM were 5% and 6% for the systems of Fe0/H2O and Fe0/UV, respectively. In contrast, the Fe/H2O2 and Fe0/UV/H2O2 systems were capable of dechlorinating TCM reductively and oxidatively by 14% and 15%, respectively. The presence of UV light was found to retard the dissolution of Fe2+, but it enhanced the rate of chloride buildup, based on the comparison of Fe0/H2O and Fe0/UV systems. In addition, WV irradiation plays only a minor role in the Fe0/UV/H2O2 system, in view of TCM dechlorination. Application of small amount of H2O2 results in the increase of Fe2+ accumulation rate in the Fe0/H2O2 system. CONCLUSIONS: TCM was dechlorinated mostly through post Fenton oxidation; reductive reaction represents a less efficient way to dechlorinate TCM. The efficiencies of overall TCM dechlorination for the two systems of Fe0/H2O2 and Fe0/UV/ H2O2 are comparable to each other, and this implies that the presence of UV irradiation imposes no significant enhancement. RECOMMENDATIONS AND OUTLOOKS: It is highly recommended to initiate effective redox dechlorination of TCM with the system of Fe0/H2O2, where the H2O2 in excess is applied at a later reaction time point. PMID- 15341316 TI - Phytoremediation of petroleum hydrocarbons in tropical coastal soils. I. Selection of promising woody plants. AB - GOAL, SCOPE AND BACKGROUND: This glasshouse study is aimed at evaluating tropical plants for phytoremediation of petroleum hydrocarbon-contaminated saline sandy subsurface soils. Tropical plants were selected for their ability to tolerate high salinity and remove No. 2 diesel fuel in coastal topsoil prior to further investigation of the phytoremediation feasibility in deep contaminated soils. The residual petroleum-hydrocarbon contaminant at the John Rogers Tank Farm site, a former petroleum storage facility, at Hickam Air Force Base, Honolulu, Hawaii, is located in a coastal area. It lies below a layer of silt in the subsurface, in loamy sand characterized by moderate salinity and high pH. Little is known regarding the ability of tropical plants to remediate petroleum hydrocarbon contaminated subsurface soil in Hawaiian and other Pacific Island ecosystems although suitable plants have been identified and utilized for bioremediation in surface soil or marine sediments. METHODS: The experiments were conducted in long narrow pots under glasshouse conditions in two phases. A preliminary experiment was done with nine tropical plants: kiawe (Prosopis pallida), milo (Thespesia populnea), common ironwood (Casuarina equisetifolia), kou (Cordia subcordata), tropical coral tree (Erythrina variegata), false sandalwood (Myoporum sandwicense), beach naupaka (Scaevola sericea), oleander (Nerium oleander), and buffelgrass (Cenchrus ciliaris). These plants were screened for resistance to high salinity treatment (2% NaCl) and two diesel fuel levels (5 and 10 g No. 2 diesel fuel/kg soil) in separate treatments. Plants that showed good tolerance of both factors were further evaluated in a second phase for their efficacy in the phytoremediation of diesel-fuel petroleum hydrocarbons under moderate salinity treatment (1% NaCl). RESULTS: Tropical coral tree and buffelgrass were susceptible to either 2% NaCl or diesel fuel at 10 g/kg soil, but tolerant of diesel fuel at 5 g/kg soil. Kiawe, milo, kou, common ironwood, N. oleander, beach naupaka and false sandalwood were tolerant of high salinity (2% NaCl) or high diesel fuel level (10 g/kg soil). These seven plants were also tolerant of the combined adverse effects of a moderate salinity (1% NaCl) and 10 g diesel fuel/kg soil. Three trees, kiawe, milo and kou significantly accelerated the degradation of petroleum hydrocarbons in the soil spiked with 10 g diesel fuel/kg soil under a moderate salinity treatment (1% NaCl). CONCLUSION: Thus the tropical woody plants, kiawe, milo and kou showed potential for use in phytoremediation of petroleum hydrocarbons in coastal tropical soils. RECOMMENDATIONS AND OUTLOOK: Two fast growing trees, milo and kou, appeared promising for further phytoremediation evaluation in experiments that simulate the soil profile at the field site. PMID- 15341317 TI - Uptake of tributyltin into willow trees. AB - BACKGROUND: Organotins have been used world-wide as antifoulants in ship paints. Repeatedly, severe effects on aquatic species have resulted. The use of organotins for this purpose was ruled out, and dumping of contaminated harbor sludge into the sea was prohibited. Land-based dumping is seen as an alternative. OBJECTIVE: This study investigates sorption, uptake and translocation of tributyltin (TBT) to willow trees in order to evaluate phytoremediation as treatment option. The study considers the influence of pH on the plant uptake of organotins. EXPERIMENTAL SET-UP: Chemicals investigated were the weak base tributyltin chloride (TBTCl) and the neutral tributyltin hydride (TBTH). Organotins were extracted from solution and plant material with toluene, and analyzed as tin by AAS with graphite oven. The pH in solution varied from pH 4 to pH 7. The sorption to living and dead roots, stems and leaves was measured in shaking experiments. The uptake into intact trees was measured at nominal levels of 1 and 10 mg TBT/l for TBTH and TBTCl at low and high pH. RESULTS: The sorption to roots and leaves dropped for dead tissue, but did not vary much with pH. The sorption to stems increased for dead stems and with pH. The solubility of TBTCl in water was below 10 mg/l and lowest at pH 4. Concentrations of TBTCl and TBTH in solutions with trees dropped rapidly to low values. Highest TBT contents in trees were found in roots and lower stems. The concentrations followed the concentrations in solution. The pH had only a small effect on the plant uptake of TBTCl, and no effect on the uptake of TBTH. No effective translocation to higher stems or leaves was found. DISCUSSION: An ion trap mechanism that accumulates the weak base TBTCl in the xylem sap of plants and leads to upward translocation could not be detected. Neither TBTCl at low or high pH, nor the neutral lipophilic chemical TBTH, were translocated effectively to leaves. The TBT+ cation sorbed strongly to plant tissue. The exact mechanism for the strong sorption of the cation is unknown, but similar effects have been observed for algae, liposomes and isolated biomembranes. CONCLUSIONS: Both the uptake of the neutral TBTH and the uptake of the neutral molecule form of TBTCl into willows was as is to be expected from theory. The cation TBT+ showed an unexpected behavior which has been observed before. No ion trap occurs, and the phytoextraction of TBT is not feasible. OUTLOOK: Planting trees, or other appropriate vegetation, could have a beneficial remediation effect by aeration of the TBT-contaminated soil or sludge. In a follow-up paper, the toxicity of TBT to willow trees will be described. PMID- 15341319 TI - State of environment in the Dniester river basin (West Ukraine). PMID- 15341318 TI - Dendroremediation of trinitrotoluene (TNT). Part 1: Literature overview and research concept. AB - BACKGROUND, AIM AND SCOPE: For decades, very large areas of former military sites have been contaminated diffusely with the persistent nitroaromatic explosive 2,4,6-trinitrotoluene (TNT). The recalcitrance of the environmental hazard TNT is to a great extent due to its particulate soil existence, which leads to slow but continuous leaching processes. Although improper handling during the manufacture of TNT seems to be a problem of the past in developed countries, environmental deposition of TNT and other explosives is still going on unfortunately, resulting from thousands of unexploded ordnance or low order explosions at munitions test areas and at current battlefields. OBJECTIVE: Sustainable phytoremediation strategies for explosives in Germany, which intend to use trees to decontaminate soil and groundwater ('dendroremediation'), have to consider that most of the former German military sites are already covered with woodlands, mainly with conifer stands. Therefore, parallel investigation of the remediation potential is necessary for both of the selected hybrids of fast growing broadleaf trees, which are waiting for planting and forest conifers, which have already proven for decades that they are able to grow on explosive contaminated sites. MAIN FEATURES: A short literature review is given regarding phytoremediation of TNT with herbaceous plants and some general aspects of dendroremediation are discussed. Furthermore, an overview of our TNT-dendroremediation research network is introduced, which has the strategic goal to make dendroremediation more calculable for a series of potent trees for site-adapted in situ application and for the assessment of tree remediation potentials in natural attenuation processes. RESULTS AND DISCUSSION: Some of our methods, results and conclusions yet unpublished are presented. For a preliminary calculation of area-related annual TNT dendroremediation potential of five-year-old trees, the following values were assessed: Salix EW-13 6.0, Salix EW-20 8.5, Populus ZP-007 4.2, Betula pendula 5.2, Picea abies 1.9 and Pinus sylvestris 0.8 g m(-2) a(-1). For a 45-year-old spruce forest, an annual natural attenuation potential of 4.2 g TNT m(-2) a(-1) was found. CONCLUSION, RECOMMENDATIONS AND PERSPECTIVE: Our main results deliver quantitative proposals for dendroremediation strategies in situ and provide decision aids. Also aspects of growth of raw materials for energy production are considered. Our dendroremediation research concept for TNT and its congeners can be easily completed for other trees of interest and it can also be applied to herbaceous plants. Knowing the current bottlenecks of phytoremediation and considering the known environmental behaviour of other contaminants, elements of our methodological approach may be easily adapted to those pollutant groups, e.g. for pesticides, pharmaceuticals, PAHs, chlorinated recalcitrants and, with some restrictions, to inorganics and to multiple contaminations. Our dynamical dendrotolerance test systems will help to predict tree growth on polluted areas. To provide some light into the black box of TNT dendroremediation, experimental data regarding the uptake, distribution and degradation of [14C]-TNT in mature tree tissues will be reported in the second part of this publication. PMID- 15341320 TI - [Replication strategies of the Flaviviridae family]. PMID- 15341321 TI - [Oral streptococci: factors and mechanisms of pathogenicity (part II)]. PMID- 15341322 TI - [Oral streptococci: implications in infectious pathology]. PMID- 15341323 TI - [Evaluation of the sensitivity to meropenem of some bacterial strains with nosocomial potential]. AB - This study analyzes the sensitivity to meropenem of 880 bacterial strains isolated from hospitalized patients in various medical units of County Hospital No. 1 Timisoara. As a result, a high sensitivity of gram-negative pathogens to meropenem (82.01%) has been noticed. Among Enterobacteriaceae, 93.17% of Escherichia coli and 71% of Klebsiella sp. were susceptible to meropenem. For Pseudomonas aeruginosa strains, a high susceptibility was noticed to meropenem (91%), more than 70% to imipenem, but piperacillin + tazobactam had 42% resistance rates. PMID- 15341324 TI - [Resistance phenotypes isolated in a department of urology]. PMID- 15341325 TI - [Bacterial strains isolated in high nosocomial risk departments]. AB - We collected 312 samples from hospitalized patients in two hospitals in Timisoara between September-December 2003. We isolated 83 strains with nosocomial potential. Identification of the germs was performed using the automatic API system, and the susceptibility tests were performed using disc-diffusion and the agar dilution test. By analyzing the extended antibiograms we categorized the germs considering their phenotypes of resistance and we remarked a high percentage of E. coli, Klebsiella pneumoniae pneumoniae and S. aureus with multiple resistance to antibiotics. PMID- 15341326 TI - [Incidence and sensitivity to antibiotics of germs isolated from surgical wound infections]. AB - We studied 119 bacterial strains isolated from postoperative infected wounds. All these strains were considered to be strains that derived from the same number of nosocomial infections, all the postoperative infections of the wounds being nosocomial infections. Regarding their frequency, we isolated the following strains: E. coli--68 strains (57%), S. aureus--37 strains (31%), Pseudomonas spp. -9 strains (8%) and Proteus spp.--5 strains (4%). We performed the bacteriological study of these strains and some correlations between them and the surgical diagnosis. In order to help the surgical therapy of the overinfected wounds, we tested the drug sensitivity of all these strains. The results regarding the drug sensitivity show that these strains have different types of resistance to antibiotics, the Pseudomonas strains being the most resistant. We noticed a continuous decrease of the sensitivity of the isolated strains to certain antimicrobial drugs, and this focuses the attention on the necessity of monitoring the antibiotic prescriptions in hospitals. Taking into account the results obtained after this study, we should reconsider the concept of nosocomial infections control through prevention activities, in order to reduce the incidence and to identify the potential causes that can lead to nosocomial infections. PMID- 15341327 TI - [The sensitivity of some strains of Klebsiella spp. collected in a neonatal department]. AB - In order to analyse the evolution of the sensitivity to antibiotics of same strains with nosocomial potential such as Klebsiella isolated in a hospital, we took into study a number of 976 samples, collected in a new-born care department of the "Dr. D. Popescu" hospital Timisoara. The study took place between November December 2002. The collected samples were pharyngeal swabs, nasal swabs, gastric aspirates, ocular and umbilical secretions, vernix, urines, faeces and blood. From all these samples 803 strains with nosocomial potential were isolated, 84 strains being Klebsiella spp. Most of the isolated strains presented multiple phenotypes of resistance to antibiotics. PMID- 15341328 TI - [Use of antimicrobial agents in the intensive care units of Bucharest hospitals]. PMID- 15341329 TI - [Discordancies between classical and API 20E microtest biochemical identification of Vibrio and Aeromonas strains]. AB - The present study reveals some discordancies of diagnosis considering Vibrionaceae strains when tested by comparison with API 20E (BioMerieux) system and classical biochemical reactions, aspects already mentioned in the literature by other authors. If any misidentification has not an important impact (e.g. identification of Aeromonas schubertii as A. sobria), others as misidentification of Aeromonas as Vibrio strains or the failure to recognize Vibrio strains raise major epidemiological problems. The present study was performed on 25 aquatic strains, comparatively identified by means of API 20E system and classical biochemical tests (using media prepared in Vibrio Laboratory of Cantacuzino Institute). All 25 strains were identified as Vibrio fluvialis by API 20E system, while using the classical tests one strain proved to belong to Pseudomonadaceae (O/F test of oxidative type), one was identified as Aeromonas schubertii, 2 Aeromonas hydrophila and 4 Aeromonas caviae. Discordancies between API 20 E system and classical tests were registered for: gelatinase (20%), LDC 12% and ADH, ODH, O/F test, saccharose, manite, inosite (4% each). The misidentification of Aeromonas as Vibrio strains by API 20E hinders the recognition of Aeromonas infections implicated more and more frequently in human pathology in immunodeficient as well as in immunocompetent hosts. Our results demonstrate that the use of API 20E system has some limits in the diagnostic of some bacterial genera (Vibrio, Aeromonas), and the introduction of these systems in the current laboratory practice requires previous information and studies regarding their efficiency and availability for the diagnostic of particular bacterial genera and species. PMID- 15341330 TI - [Comparative study of classical and commercial microtest API galleries in the diagnosis of cholera infection]. AB - Cholera still remains in the top of causes generating global mortality and morbidity, as revealed by the latest WHO reports. In 2000, on CDC/Atlanta website, the cholera agent was mentioned as potential biological weapon for bioterrorism actions in 9460 publications. Considering these aspects, a correct and rapid diagnosis is necessary in order to take appropriate epidemiological measures and to prevent the secondary transmission. Our study evaluated the efficiency of microtest systems API20E, API 20 NE and RapID 20E, by comparison with classical biochemical tests concerning the diagnosis of 150 Vibrio cholerae strains isolated during cholera epidemics of Romania (1990-1995). Our results demonstrated that classical biochemical identification still remains the most secure diagnostic method for Vibrio cholerae strains. The kits API 20NE gave the highest number of results in concordance with the classical tests. For these reasons, the confirmation of cholera infection must be performed in a National Reference Center. The limits of API galleries concerning the diagnosis of Vibrio cholerae strains must be taken into account and in case of suspect clinical symptoms, the isolated strains must be sent to the Cantacuzino Institute, even if the microtest galleries have not identified V. cholerae strains. PMID- 15341331 TI - [Effect of transferrin on the expression of virulence factors in Listeria monocytogenes strains]. AB - Iron is an essential element for the great majority of microorganisms, which developed transport systems for iron acquisition, because during the colonization and invasion processes the microorganisms encounter iron-limiting conditions. The bacterial genes implicated in the iron transport and those codifying for other virulence factors are simultaneously depressed. The purpose of this study was to investigate the role of transferrin on the virulence factors expression in food isolated Listeria monocytogenes strains. Our results showed that transferrin stimulates the bacterial growth rates, as well as the adherence and invasion capacity (27 x 10(4) CFU/ml vs 13 x 10(4) CFU/ml) to cellular substrate and inert one (as shown by slime test). Transferrin also induced the secretion of haemolysins, amylases and lecithinases, demonstrating the role of iron ions in the virulence genes derepression and the simulation of bacterial growth rate. PMID- 15341332 TI - [Correlations between the level and frequency of blood eosinophilia and hydatid cyst location]. PMID- 15341333 TI - [Antibiotic resistance of some Pseudomonas aeruginosa isolates]. PMID- 15341334 TI - [Leptospirosis--epidemic, clinical and diagnostic aspects]. AB - Leptospirosis is a major public health problem throughout the world. We studied 22 cases of leptospirosis. They were hospitalised between 1999-2003. Men were more affected than women and those with rural origin prevailed, too. The direct infection was more often found in these cases. These patients presented a large spectrum of signs and symptoms attributable to this spirochetal pathogen. The etiology was dominated by L. pomona, L. canicola, L. wolffi. Leptospirosis presented with 3 syndromes: Weil's syndrome, atypical pneumonia syndrome and aseptic meningitis. The adequate treatment was with penicillin or ampicillin i.v. PMID- 15341335 TI - [Mechanisms of onset of quinolone-resistant mutants in E. coli strains]. PMID- 15341336 TI - [Sepsis--etiology and antibiotic resistance]. PMID- 15341337 TI - [Clinical and biological aspects of liver damage in infectious mononucleosis]. AB - Liver damage, clinical manifest or inapparent is a frequent manifestation of infectious mononucleosis. This study has in view the forms and frequency of manifestation of liver damage in infectious mononucleosis. The retrospective study has included 115 patients with infectious mononucleosis, hospitalised in Brasov Infectious Diseases Hospital between 1.01.1998 and 31.12.2002. The enlargement of the liver as unique manifestation in liver damage was noticed in 27% of the cases; jaundice of the skin and sclerae in 10.4% of the cases; infectious mononucleosis hepatitis in 55.6% of the cases; and in 11.3% of cases high serum levels of GPT were noted. The levels of serum protein were also affected, low-grade increase of protein serum levels--18.2% of the cases, hypoalbuminemia--82.4% of cases and subunitary albumin/globulin ratio in 70.3% of the cases. All these levels might be indicators for liver damage. PMID- 15341338 TI - Will Medicare Advantage rate hike bring providers back to cap? PMID- 15341339 TI - Does capitated reimbursement reduce the rate of specialty procedures? PMID- 15341340 TI - Drug trend slows in 2003, but higher utilization still driving costs. PMID- 15341341 TI - Capitation increasingly funds behavioral health treatment for youths. PMID- 15341342 TI - Handing ED physicians the cath lab's keys speeds procedures. PMID- 15341343 TI - BlueWorks aims to improve performance throughout Blues' system. AB - Last October, the Chicago-based Blue Cross and Blue Shield System, in collaboration with researchers at Harvard Medical School in Boston, embarked on a unique collaboration to identify and promote the most promising programs developed by Blues companies to maintain health care affordability while encouraging the adoption of effective clinical practices. Those efforts are beginning to bear fruit, with the first six winning initiatives revealed in the first BlueWorks Quarterly Report. PMID- 15341344 TI - Wisconsin hospitals share performance data online. AB - Performance data is no longer a closely-held secret, as thousands of hospitals are reporting this information to the CMS and other quality improvement initiatives. The Wisconsin Hospital Association has tapped into this flow of data to build a website that allows hospitals in that state to compare their performance directly to other facilities. PMID- 15341345 TI - HealthPartners uses specialist scorecard to improve clinical outcomes. AB - In the world of managed care, primary care physicians used to be the coordinators of care and they determined when patients saw specialists. But that view was turned upside down by a patient backlash against the gatekeeper approach. To keep their customers, managed care companies decided to allow direct access to specialists. PMID- 15341346 TI - Teaching undergraduates how to write and submit a research manuscript. AB - Undergraduate nursing students have long been excluded from performing original theory-based nursing research due to severe time and work constraints. However, without actual research opportunities, students also lack research-writing experiences. This author describes successful strategies for teaching an undergraduate class how to write and submit a manuscript that describes an original research project. PMID- 15341347 TI - Side steps to publication. AB - After you have prepared an inservice presentation, a term paper, or a project, you often have the building blocks for a publishable paper. You have organized the content and done the research--now you need to translate your paper or oral presentation into the journal format. This experienced author offers tips for successfully building your article on this foundation. PMID- 15341348 TI - Relationship of subgingival and salivary microbiota to gingival overgrowth in heart transplant patients following cyclosporin A therapy. AB - BACKGROUND: Severe gingival overgrowth (GO) is induced in patients taking cyclosporin A (CsA) following organ transplantation. Determining which patient will develop GO is still not possible. The purpose of this study was to establish an association between CsA and gingival overgrowth in heart transplant patients taking into account periodontal and microbiological conditions. METHODS: Thirty patients (10 female, 20 male; range: 13 to 67 years; mean age: 44.89) undergoing CsA treatment were evaluated using the gingival index (GI), plaque index (PI), probing depth (PD), and clinical attachment level (CAL). Subgingival samples collected from the deepest site of each quadrant and saliva samples were submitted to microbial analysis. All patients had at least 12 teeth. Exclusion criteria were the use of antibiotics and/or having undergone periodontal treatment 6 months prior to the study. Patients were divided in two groups: with gingival (GO+) and without gingival overgrowth (GO-). RESULTS: There were no statistically significant differences between the GO+ and GO- groups when CsA dosage, time since transplant, GI, PI, PD, and CAL were compared. Microbiological examination of the subgingival samples detected the following microorganisms: Actinobacillus actinomycetemcomitans (23%), Porphyromonas gingivalis (36%), Prevotella intermedia (93%), Fusobacterium sp. (66%), Campylobacter rectus (30%), Micromonas micros (66%), enteric rods (0%), and yeasts (30%). A positive association between M. micros and the GO+ group was found (P < 0.001). Yeasts were detected in 30% of the subgingival and saliva samples. CONCLUSIONS: Clinical parameters were not sufficient to determine which patients would develop GO. However, colonization by M. micros might play a role in the etiology of GO. PMID- 15341349 TI - Biomimetic approach on human periodontal ligament cells using synthetic oligopeptides. AB - BACKGROUND: Periodontal ligament (PDL) cells, connecting root cementum with alveolar bone, are important for periodontal wound healing. In order to obtain a predictable periodontal regeneration, selective adhesion and proliferation of PDL cells are essential. The purpose of this study was to investigate the effects of synthetic peptides mimicking cell-binding domain of fibronectin (FN) on human PDL cells. METHODS: Two types of oligopeptides, Gly3-Pro-His-Ser-Arg-Asn-Gly6-Arg-Gly Asp-Gly (G3PHSRNG6RGDG) and Gly3-His-Pro-Asn-Arg-Ser-Gly6-Arg-Gly-Asp-Gly (G3HPNRSG6RGDG), were constructed using a solid-phase peptide synthesizer. Fibronectin type III ninth to tenth domain (FN III 9-10) and Arg-Gly-Asp-Ser (RGDS) were prepared for comparison with the effects of synthetic peptides. These peptides were coated onto 96-well cell culture plates with 0.001 approximately 100 microM concentrations. Cultured human PDL cells were then applied to the peptide-coated wells at a density of 1 x 10(4)/well. After 1 hour incubation at 37 degrees C, adhered cells were fixed, stained, and examined by phase contrast microscopy for cell spreading assay. Attached PDL cells were solubilized with 2% sodium dodecyl sulfate (SDS) for the cell attachment assay by measuring absorbance at 595 nm in microplate reader. Western blot analysis was performed to determine extracellular signal-regulated kinase (ERK1/2) activity. RESULTS: Cell attachment and spreading assays revealed that G3PHSRNG6RGDG (> or = 10 microM) possesses similar adhesive behavior to FN III 9-10. G3PHSRNG6RGDG showed a comparable ERK1/2 activity when compared to FN III 9-10. CONCLUSIONS: G3PHSRNG6RGDG enhanced an attachment and spreading of human PDL cells thereby increasing ERK1/2 activity. Taken together, it is anticipated that this peptide might be a potential tool for arranging a biologically attractive environment for PDL cells, which would enhance periodontal regeneration efficacy. PMID- 15341350 TI - Mast cell subpopulations in gingival overgrowth induced by immunosuppressive and nifedipine medication. AB - BACKGROUND: An immunohistochemical study was conducted to compare distributions of mast cell subpopulations in normal human gingiva and in gingival overgrowth induced by nifedipine and immunosuppressive medication. METHODS: Gingival samples were collected from 12 triple-medicated organ transplant recipients (immunosuppression group), 11 triple-medicated organ transplant recipients taking nifedipine (immunosuppression plus nifedipine group), 11 nifedipine-medicated cardiac outpatients (nifedipine group), and 20 generally healthy individuals (control group). Cryostat sections were stained with mAbs for tryptase and chymase, and an avidin-biotin enzyme complex method was used to detect tryptase positive mast cells (MC(T)), tryptase- and chymase-positive mast cells (MC(TC)), and chymase-positive mast cells (MC(C)). Total numbers of labeled cells were determined in connective tissue beneath the sulcular epithelium, connective tissue beneath the oral epithelium, and middle connective tissue. Statistical analyses were conducted using the Kruskal-Wallis test, the Mann-Whitney U-test, and Pearson's correlation test. RESULTS: In the three counting zones combined, numbers of MC(TC) cells and MC(C) cells were lower (P = 0.001 and P = 0.048, respectively) in the immunosuppression group than in the control group. The difference in numbers of MC(TC) cells was most marked in the middle connective tissue. Nifedipine medication had no effect on numbers of the mast cell subclasses. CONCLUSIONS: Immunosuppressive medication without concomitant nifedipine decreases the numbers of MC(TC) and MC(C) in overgrown gingiva. Chymase-positive mast cells may play a role in formation of gingival overgrowth, especially in patients receiving cyclosporin A (CsA) medication with no concomitant nifedipine. In this respect, nifedipine and CsA are different. PMID- 15341351 TI - Inhibition of osteogenesis in vitro by a cigarette smoke-associated hydrocarbon combined with Porphyromonas gingivalis lipopolysaccharide: reversal by resveratrol. AB - BACKGROUND: Smoking and infection with Gram-negative bacterial pathogens are risk factors for alveolar bone loss. The aims of this study were: 1) to examine the combined effects of an aryl hydrocarbon, benzo[a]pyrene (BaP), that is concentrated in cigarette smoke, and lipopolysaccharide (LPS) extracted from Porphyromonas gingivalis on osteogenesis in a rat bone marrow cell (RBMC) model of osteogenesis; and 2) to determine whether resveratrol (Res), an aryl hydrocarbon receptor antagonist, could reverse the putative inhibitory effects of BaP + LPS on osteogenesis. METHODS: LPS of P. gingivalis strain 2561 was introduced in various concentrations to the RBMC in 96-well plates and kept in culture for 8 to 12 days. The same protocol was used for studying BaP and LPS + BaP combinations. Following the incubation periods, parameters of osteogenesis were measured, including formation of mineralized bone nodules, alkaline phosphatase activity, and total cell protein. Transcription of the pro inflammatory cytokine interleukin (IL)-1beta in the cultures was determined by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Bone nodule formation generally decreased significantly with increasing LPS concentrations (P<0.05), whereas total cell protein decreased only slightly (P>0.05). BaP in previously high concentrations alone also caused a significant dose-dependent decrease in bone nodule formation (P<0.05) but when half maximal doses were used, significant decreases were most often seen when LPS was added. Hence, in combination, the inhibitory effects of LPS + BaP on osteogenesis were additive, inhibiting bone nodule formation up to 9-fold. Resveratrol partially reversed the inhibitory effects of low concentrations of LPS alone, and completely reversed the inhibition of nodule formation when low concentrations of LPS were combined with BaP. IL-1beta expression generally fluctuated inversely to the inhibitory activity of LPS, LPS + BaP, and LPS + BaP + Res combinations. CONCLUSIONS: Smoke derived aryl hydrocarbons and bacterial LPS may act additively to inhibit bone formation. The findings may explain, in part, why net periodontal bone loss is greater and bone healing is less successful in smokers than non-smokers with periodontal infections. Reversal of the inhibitory effects in vitro by resveratrol suggests that this phytoalexin should be studied further for its potential therapeutic value, given its aryl hydrocarbon receptor antagonism and apparent anti-inflammatory activity. PMID- 15341352 TI - A comparative study of coronally advanced flaps with and without the addition of enamel matrix derivative in the treatment of marginal tissue recession. AB - BACKGROUND: The primary aim of this randomized, controlled, clinical investigation was to evaluate the differences in clinical parameters of root coverage procedures utilizing coronally advanced flaps (CAF) with and without enamel matrix derivative (EMD). Another aim was to evaluate if EMD can influence keratinization of tissues utilizing these procedures. METHODS: Fifty-eight contralateral sites in 17 patients with > or = 2 mm of Miller Class I, II, and III buccal marginal tissue recession were treated. Test sites received CAF plus EMD while control sites were treated with CAF only, with each patient serving as his own control. Patients were followed for 3 and 6 months. All data were analyzed using the method of generalized estimating equations. A P value <0.05 was considered significant (alpha = 0.05). RESULTS: There was a mean increase in keratinized tissue of 0.60 mm for test sites and a mean decrease of 0.05 mm for control sites. Test sites demonstrated significantly better root coverage (P<0.001), 89.7% and 92.9% root coverage after 3 months and 6 months, respectively, while control sites had 56.6% and 66.8% root coverage after 3 months and 6 months, respectively. There was significantly more root coverage among test sites compared to control sites, regardless of arch or Miller classification. CONCLUSIONS: Based on the results of this study, the application of enamel matrix derivative to denuded root surfaces receiving coronally advanced flaps significantly increased the percentage of root coverage compared to CAF without EMD. In addition, EMD application was accompanied by a significant increase in keratinized tissue 6 months after surgery. PMID- 15341353 TI - Guided tissue regeneration with bioabsorbable barriers. II. Long-term results in infrabony defects. AB - BACKGROUND: The aim of this 5-year randomized controlled clinical trial was to evaluate the long-term results after guided tissue regeneration (GTR) therapy of infrabony defects using two bioabsorbable barriers. METHODS: Fifteen pairs of contralateral infrabony defects in 15 patients with moderate to severe periodontitis were treated. Each patient received one polydioxanon (test: T) and one polylactide acetyltributyl citrate (control: C) barrier by random assignment. At baseline, 12, and 60 +/- 3 months after surgery clinical parameters and standardized radiographs were obtained. Vertical bone levels (PBL-V) were measured during surgery and 60 +/- 3 months later by transgingival bone sounding. RESULTS: Thirteen patients were available for the 60-month examinations. Twelve and 60 +/- 3 months after GTR, statistically significant (P< or =0.001) vertical attachment (CAL-V) gain was found in both groups (T12: 3.5 +/- 1.5 mm; T60: 2.2 +/- 1.8 mm; C12: 4.0 +/- 0.9 mm; C60: 2.4 +/- 1.0 mm). However, from 12 to 60 months after therapy both groups experienced significant CAL-V loss (P<0.05): two defects in the test group and three in the control group had CAL-V loss >2 mm compared to the 12-month reexamination. Twelve and 60 +/- 3 months after surgery, statistically significant (P<0.05) radiographic bony fill was found in both groups (T12: 1.2 +/- 1.3 mm; T60: 1.5 +/- 2.2 mm; C12: 0.9 +/- 1.4 mm; C60: 1.0 +/- 1.6 mm). Further, 60 months after surgery significant (P<0.05) PBL-V gain was found in both groups (test: 1.8 +/- 2.3 mm; control: 2.2 +/- 1.8 mm). The study failed to show statistically significant differences between test and control regarding CAL-V and PBL-V gain 60 months after surgery. CONCLUSION: CAL-V gain achieved after GTR therapy in infrabony defects using both bioabsorbable barriers was stable after 5 years in 21 of 26 defects (81%). PMID- 15341354 TI - Periodontal treatment with an Er:YAG laser compared to ultrasonic instrumentation: a pilot study. AB - BACKGROUND: The aim of the present study was to compare the effectiveness of an Er:YAG laser to that of ultrasonic scaling for non-surgical periodontal treatment. METHODS: Twenty patients with moderate to advanced periodontal disease were randomly treated in a split-mouth design with a single episode of subgingival debridement using either an Er:YAG laser device (160 mJ/pulse, 10 Hz) combined with a calculus detection system with fluorescence induced by 655 nm InGaAsP diode laser radiation (ERL), or an ultrasonic instrument (UI). Clinical assessments of full-mouth plaque score (FMPS), bleeding on probing (BOP), probing depth (PD), gingival recession (GR), and clinical attachment level (CAL) were made at baseline and at 3 and 6 months following therapy. RESULTS: No differences in any of the investigated parameters were observed at baseline between the two groups. The mean value of BOP decreased in the ERL group from 40% at baseline to 17% after 6 months (P<0.0001) and in the UI group from 46% at baseline to 15% after 6 months (P<0.0001). The sites treated with ERL demonstrated mean CAL gain of 1.48 +/- 0.73 mm (P<0.001) and of 1.11 +/- 0.59 mm (P<0.001) at 3 and 6 months, respectively. The sites treated with UI demonstrated mean CAL gain of 1.53 +/- 0.67 mm (P<0.001) and of 1.11 +/- 0.46 mm (P<0.001) at 3 and 6 months, respectively. No statistically significant differences were observed between the groups (P>0.05). CONCLUSION: Within the limits of the present study, it can be concluded that both therapies led to significant improvements of the investigated clinical parameters. PMID- 15341355 TI - Substance P enhances the inhibition of osteoblastic cell differentiation induced by lipopolysaccharide from Porphyromonas gingivalis. AB - BACKGROUND: Substance P (SP) is a multifunctional neuropeptide that transmits pain signals, regulates the immune system, and may modulate emotional stress. SP stimulates bone resorption activity of osteoclasts, and SP level in gingival crevicular fluid is correlated with the degree of periodontal inflammation. However, the exact roles of SP in bone metabolism and periodontal diseases are poorly understood. To elucidate the effect of stress on bone metabolism, we investigated the effect of SP on osteoblastic cell differentiation in the presence of lipopolysaccharide from Porphyromonas gingivalis (P-LPS). METHODS: The primary osteoblastic cells were isolated from fetal rat calvaria (RC) and cultured with SP, P-LPS, and an SP antagonist (SPa). The effects of SP on bone nodule (BN), alkaline phosphatase (ALPase) activity, mRNA expressions of SP receptor, bone matrix proteins, and Cbfa 1 were investigated. RESULTS: SP stimulated the expression of SP receptor mRNA in RC cells and enhanced its expression in the presence of P-LPS (50 ng/ml). SP inhibited BN formation and ALPase activity in a dose-dependent manner (10(-7) to 10(-5) M) and further suppressed mRNA expression of bone sialoprotein, osteopontin, and osteocalcin but not of type I collagen mRNA. The inhibitory effects were enhanced in the presence of P-LPS and blocked by Spantide III. Furthermore, the expression of Cbfa 1 mRNA was also markedly suppressed in the presence of SP and P-LPS. CONCLUSION: These findings suggest that SP inhibits osteoblastic cell differentiation and may be related to bone metabolism in periodontal diseases under conditions of stress. PMID- 15341356 TI - Immediate implants supporting single crown restoration: a 4-year prospective study. AB - BACKGROUND: The placement of implants at the time of tooth extraction has several clinical advantages, such as preservation of the alveolar ridge width and height and reduction of the restorative treatment time. The aim of this study was to evaluate the cumulative success rate of dental implants placed in fresh extraction sockets with and without guided bone regeneration (GBR) used to support a single crown restoration. All of the patients were preselected as candidates for implants. METHODS: Ninety-five patients aged 20 to 68 years with 163 implants were included. All patients were partially edentulous and participated in a personally tailored recall schedule. The follow-up period was 48 months. Patients underwent a clinical and radiographic evaluation annually. RESULTS: The 4-year cumulative success rate was 97%. Five of the 163 implants failed, two during the initial healing time, which were considered early failures and three a year after prosthetic rehabilitation, which were considered late failures. No failure of prosthetic rehabilitation was observed. CONCLUSIONS: Implants placed into fresh extraction sockets with or without regenerative procedures and used to support single crown prosthesis showed a very high cumulative success rate (97%) in a 4-year prospective study. Several observations should be made: 1) all the patients were preselected as candidates for implants and were following a strict oral hygiene regimen; 2) all efforts were made to reduce the number of cases requiring GBR procedures; 3) all the implants had an acid etched/sandblasted implant surface; and 4) all the prosthetic restorations were single crowns. PMID- 15341357 TI - Comparison of keratinocyte proliferation in diabetic and non-diabetic inflamed gingiva. AB - BACKGROUND: Keratinocytes are chiefly cells of the epidermis but also constitute 90% of the gingival cells. The molecular mechanisms of proliferative activity in keratinization whereby diabetes alters periodontal physiology have not been elucidated. In this study, we aimed to investigate the role of gingival keratinocytes in hyperglycemic subjects by examining their mitotic activities. METHODS: We tested 30 patients with periodontitis, of whom 15 were type II diabetics and the remainder systemically healthy. Biopsies were obtained from the bottom of the deepest pocket in each subject by reverse beveled incision. Formalin-fixed and paraffin-embedded specimens were then processed for periodic acid-Schiff (PAS)-diastase histochemistry and proliferating cell nuclear antigen (PCNA) (P10). Immunohistochemical studies were employed to determine the presence of PCNA and were used to detect the proliferating potential of keratinocytes needed in synthesizing DNA. The expression of PCNA was evaluated using an immunoperoxidase technique and PC10 monoclonal antibody to PCNA. Mitotic index was calculated from basal cells. Statistical analysis employed the chi-square test. RESULTS: No significant difference between the diabetic and non-diabetic patients was found in the mitotic index of the oral-gingival epithelium. CONCLUSION: Although the mitotic index in patients with diabetes was slightly lower, keratinization in the gingival tissues for both groups was essentially identical. PMID- 15341358 TI - Matrix metalloproteinase-2 may be involved with increased bone loss associated with experimental periodontitis and smoking: a study in rats. AB - BACKGROUND: Smoking has been associated with periodontitis severity and is considered a risk factor for its development. It has been reported that matrix metalloproteinase (MMP) produced by host cells plays a major role in periodontal tissue destruction. Thus, the present study tested, in rats, the hypothesis that local increased levels of MMP-2 would be associated with the enhanced periodontitis-related bone loss after intermittent cigarette smoke inhalation (CSI). METHODS: Twenty-seven adult male Wistar rats were used. A ligature was placed around one of the mandibular first molars of each animal and they were randomly assigned to the following control (N = 13) or CSI (N = 14) group. Sixty days later, the animals were sacrificed, the gingival tissues harvested, and the specimens processed for decalcified sections. Extracts from the gingival tissues were prepared and assayed for MMP-2 expression. RESULTS: Intergroup comparisons (unligated sites) showed that CSI might directly affect alveolar bone (0.16 +/- 0.03 mm2 versus 0.24 +/- 0.09 mm2 for non-smokers and smokers, respectively; P = 0.001). Moreover, CSI significantly enhanced bone loss resulting from experimental periodontitis (0.64 +/- 0.36 mm2 versus 1.50 +/- 0.50 mm2 for non smokers and smokers, respectively; P<0.05). In addition, zymography demonstrated that CSI also enhanced both MMP-2 levels and activity in the gingival tissues around ligated teeth. CONCLUSION: Within the limits of the present investigation, it can be assumed that the effect of CSI on MMP-2 levels and activity may account for the increased periodontitis progression rate observed in smokers. PMID- 15341359 TI - A multicenter study evaluating the sensitization potential of enamel matrix derivative after treatment of two infrabony defects. AB - BACKGROUND: Several studies reported some success toward regeneration in infrabony defects using enamel matrix derivative (EMD). Clinically and statistically significant improvements in probing depth reduction, clinical attachment levels, and bone fill have been demonstrated. This multi-center study evaluated the potential for sensitization to EMD in a subgroup of periodontal patients treated at least twice with at least 2 months between treatments. METHODS: Three hundred seventy-six (376) patients in 11 university-based postgraduate periodontics programs and five private practices were selected. Surgeries were performed on infrabony defects. Following reflection of mucoperiosteal flaps and debridement of the root surface and defect, root conditioning (either citric acid pH = 1 or 24% EDTA) was performed and the site was irrigated with sterile saline. Enamel matrix derivative was reconstituted and applied to the exposed root surface and the bony defect. Flaps were sutured and pressure applied for 5 minutes. The second test defect was treated in a similar manner at least 8 weeks after the first surgery. The patient was given a diary card where any subjective adverse events (erythema, swelling, itching, headache, root hypersensitivity, or pain) were recorded at weeks 1 and 2 post-surgery. In addition, objective adverse events (gingival inflammation, ulcers, abscess, cratering, and lesions) were recorded by the investigator on an adverse event form. RESULTS: No clinical adverse reactions to multiple applications of EMD were noted. Of 376 patients, two were referred to a dermatologist for evaluation, but neither had signs indicating any adverse events due to EMD treatment. Instead their reactions were classified as a small local abscess and tinea cruris. The single immunoassay performed (on the patient with a small local abscess) did not demonstrate any EMD-reactive antibodies, neither IgE nor IgG. Other subjective/objective reactions that occurred during this study were of the type that are commonly experienced by patients immediately following periodontal surgery, but were not related to EMD. They included headache, swelling, itching, pain, and root hypersensitivity. CONCLUSIONS: This study demonstrated a lack of clinical adverse reactions following two separate applications of EMD. Any subjective/objective adverse reactions experienced by the patient were typical complications following routine periodontal surgery and were not directly related to the use of enamel matrix derivative. PMID- 15341360 TI - Tongue-cleaning methods: a comparative clinical trial employing a toothbrush and a tongue scraper. AB - BACKGROUND: It is estimated that approximately 85% of all halitosis cases have their origin within the mouth; of these, 50% are caused by tongue residues. Previous studies have established that hydrogen sulfide and mercaptans are the primary components of halitosis. Thus, tongue cleaning gains importance as a means of halitosis management. METHODS: This investigation compared the efficacy of two mechanical methods for tongue cleaning through a handheld sulfide monitor. This crossover trial was carried out with 10 healthy subjects, 20 to 50 years old. Before the baseline measurement of the volatile sulfur compounds (VSCs), the subjects were instructed to refrain from any tongue cleaning method for 48 hours. The 10 participants were then placed in one of two groups (five each): 1) first week: tongue scraper, second week: soft-bristle toothbrush; 2) first week: toothbrush, second week: tongue scraper, with a 48-hour wash-out period between each week. RESULTS: The baseline measurements were compared with those of the end of each week using the Dunn method (alpha = 0.01). The tongue scraper showed a 75% reduction in VSCs, while the toothbrush only achieved a 45% reduction in VSCs. CONCLUSION: Although the tongue coating was removed by both methods, the tongue scraper performed better in reducing the production of volatile sulfur compounds. PMID- 15341361 TI - Bone apposition in surgical bony defects following orthodontic movement: a comparative histomorphometric study between root- and periodontal ligament damaged and periodontally intact rat molars. AB - BACKGROUND: The influence of orthodontic tooth movement with diminished periodontal support is unclear. The aim of the present study was to evaluate bone healing in surgical defects following orthodontic tooth movement with and without periodontal ligament (PDL) and root surface damage. METHODS: The study comprised 33 adult male Wistar rats, divided into two groups: group 1 (n = 14) with bony defect and no root damage and group 2 (n = 19) with periodontal bony defect including root/PDL damage on the mesial root of the maxillary first molar. One week after a surgical defect was created, orthodontic protraction of the right maxillary first molar was initiated in both groups. After 2 weeks of protraction, retention of 1 week was established; at the end of this period block sections were made. Histomorphometric analysis through light microscopy of decalcified tissue was performed. Results were statistically analyzed using independent samples t test and analysis of variance (ANOVA) with repeated measures. RESULTS: Differences between groups in total area of bone defect and bone apposition were not statistically significant. Bone apposition calculated as percentage of the bone defect was significantly (t-test) greater (P = 0.002) in group 2 (46.21%) than in group 1 (24.95%). Within each group, area of bone apposition was significantly (ANOVA) greater in the distal than in the mesial quadrants of the bony defect (P = 0.006) and in the apical than the occlusal ones (P = 0.021). CONCLUSION: Following orthodontic tooth movement, periodontal bony defects showed enhanced bony healing compared with alveolar bone defects with no direct association with the periodontal attachment apparatus. PMID- 15341362 TI - Inflammatory response to acute coronary syndrome in patients with coexistent periodontal disease. AB - BACKGROUND: The inflammatory process that occurs in atherosclerotic plaque situated in the coronary vessel often leads to its destabilization, which in turn results in clot formation and the occurrence of acute coronary syndrome. Acute coronary syndromes (unstable angina, myocardial infarction) are among the leading causes of death in the human population. An inflammatory factor which leads to this process may be bacterial pathogens originating in the periodontal tissues. The aim of the study was to evaluate the influence of periodontal disease on the intensity and dynamics of the inflammatory response in patients with acute coronary syndrome and coexistent periodontal diseases. METHODS: The study involved a group of 50 consecutive patients (nine females; 41 males) < or = 60 years of age (37 to 60; mean 51) with coronary pain and with an initial diagnosis of acute coronary syndrome admitted to the Coronary Care Unit of the Department of Cardiology. Dental examinations were carried out during the first 24 hours of hospitalization. Blood samples were taken at admission (examination 1), after 10 to 12 days of hospitalization (examination 2), 3 months after the acute coronary syndrome (examination 3), and, in some patients, 6 months after the acute episode (examination 4). This information was used to estimate the mean serum levels of interleukin-1 (IL-1) and tumor necrosis factor (TNF-alpha). RESULTS: Chronic periodontitis was found in all patients. The patients were divided into two groups on the basis of periodontal disease and clinical attachment loss (CAL). The study showed raised mean values for TNF-alpha in examinations 2, 3, and 4 in patients with more advanced periodontal disease. There was also an increase in IL 1 concentration in the acute phase and in long-term observation (examination 3) in these patients. The patients with more advanced CAL had more pronounced periodontal disease and higher bleeding index values. All patients were characterized by high mean values of plaque index (46%) and bleeding index (80%). CONCLUSIONS: The periodontal health of patients admitted to the Coronary Care Unit due to acute coronary syndrome is unacceptable. The mean values for CAL and probing depth, as well as extensive bleeding on probing sites indicate the presence of active periodontal disease, which may affect the incidence of cardiovascular disease. Although there were no significant differences in serum TNF-alpha or IL-1 levels in acute coronary disease patients with advanced periodontal disease compared to those with less advanced periodontal disease, we observed that patients with acute coronary syndrome and with more advanced periodontal disease tend to be characterized by higher mean values of serum concentrations of IL-1 in the acute phase of acute coronary syndrome, as well as in the long term, and of TNF-alpha in the long-term observation. Patients with less advanced periodontal disease were characterized by a faster diminution of the inflammatory response in comparison to the groups with more advanced periodontal disease. PMID- 15341363 TI - Root resorption in chronic periodontitis: a morphometrical study. AB - BACKGROUND: It has been previously observed that teeth with severe chronic periodontitis experience more transient external root resorptions than teeth with less severe disease. METHODS: Sixty-six teeth diagnosed with different levels of chronic periodontitis in 48 patients were observed with a light microscope. Main characteristics of teeth with root resorptions, as well as percentages of area and volume resorbed on each radicular third of the teeth, were determined. RESULTS: Over 80% (53) of the teeth exhibited resorption: 46.67% in group 1 (slight periodontitis); 85% in group 2 (moderate periodontitis); and 93.55% in group 3 (severe periodontitis). In group 1, the average resorption length was 295.65 microm and the average area, 11,212.45 microm2. In group 2 these measures were 519.09 and 25,678.102, respectively. In group 3, the average length was 675.95 microm, and the average area 36,925.06 microm2. Except for the resorbed volume on the apical third, the other resorption percentages were higher in group 3 (severe periodontitis) and smaller in the group 1 (slight periodontitis), but the location of the largest resorption percentages varied among the three groups. CONCLUSIONS: The severity of periodontitis plays a major role in the presence and extent of transient external root resorption. In the three groups, most of the teeth showed resorptions on the apical third, but severity of periodontitis determined the location of the largest percentages of area and volume resorbed. PMID- 15341364 TI - Periodontal attachment loss in an urban population of Brazilian adults: effect of demographic, behavioral, and environmental risk indicators. AB - BACKGROUND: There is little information about the occurrence and risk factors of periodontal diseases in developing countries. This study describes the clinical attachment loss (CAL) in an adult Brazilian population and performs a risk assessment of demographic, behavioral, and environmental exposures. METHODS: A representative sample of 853 dentate individuals (age: 30 to 103 years) was selected by a multistage probability sampling method. The subjects had a full mouth clinical examination of six sites per tooth and were interviewed using a structured written questionnaire. RESULTS: Seventy-nine percent (79%) and 52% of the subjects and 36% and 16% of the teeth per subject had CAL > or = 5 and > or = 7 mm, respectively. A multivariable model showed that 40 to 49 and > or = 50 years olds had 3.0 and 5.9 times higher risk for moderate CAL and 7.4 and 25.4 times higher risk for severe CAL, compared to the 30 to 39 years olds. Moderate cigarette smokers had a significantly higher risk for moderate (relative risk ratio [RRR] = 2.1) and severe CAL (RRR = 3.4), and heavy smokers had a higher risk for moderate (RRR = 3.0) and severe CAL (RRR = 8.2) compared to non-smokers. A significantly higher risk for severe CAL was also present in males (RRR = 1.6), subjects with low (RRR = 1.8) or medium socioeconomic status (RRR = 1.6), and those with a history of irregular dental visits (RRR = 2.1). Diabetic status and race did not show significant associations with CAL after adjusting for other effects. CONCLUSIONS: This Brazilian population had a high occurrence of attachment loss. A population-based strategy that includes the establishment of prevention and health promotion programs targeting high-risk groups is highly desirable for controlling the high occurrence of attachment loss in this population. PMID- 15341365 TI - Re: The effect of menstrual cycle on periodontal health. PMID- 15341366 TI - Complications associated with implant surgical procedures part II: Treatment. PMID- 15341367 TI - Avian influenza--current evaluation of risks to humans from H5N1 following recent reports. PMID- 15341368 TI - Global Advisory Committee on Vaccine Safety, 10-11 June 2004. PMID- 15341369 TI - The school nurse--every school should have one. PMID- 15341370 TI - Special needs children are special. PMID- 15341371 TI - Children's sleep: problems and solutions. AB - It is now believed that up to 43% of children can experience a sleep problem at some stage during their development. Reduced quality of sleep can have important implications for the developing child as it can impair growth, learning and emotional development. It is important for health professionals to understand the significance of sleep and the ways to alleviate problems so that these can be tackled at an early age. By educating people in how to get a good night's sleep we shall hopefully move towards a more happy, healthy and capable child population. PMID- 15341372 TI - Home and dry--some toilet training tips to give parents. PMID- 15341373 TI - What do we know about... Oral health for families? AB - The author outlines improvements in Britain's dental health and summarises practical advice to pass on to families with young children and adults of all ages. The importance of fluoride and moderating the frequency of snacks is emphasised. Current thinking on diet and dental decay is discussed, including new developments in our understanding of the part played by carbohydrates, which is less simplistic than previously thought. PMID- 15341374 TI - What do mothers of young children think of community pharmacists? A descriptive survey. AB - AIM: To discover the attitudes of mothers of young children towards community pharmacists, and mothers' use of community pharmacies for childhood illnesses OBJECTIVES: To discover how frequently mothers visit pharmacies; for which childhood complaints they tend to consult a pharmacist first, rather than other health professionals; to identify how helpful mothers feel their pharmacists are; and to identify whether mothers prefer pharmacists with particular demographic characteristics, eg having children of their own. METHODS: A questionnaire survey based on the results of four focus groups conducted with mothers of children aged under five years. 117 questionnaires were distributed by five pharmacies in Northern England to a convenience sample between July and September 2000, with one written reminder to non-respondents one month later. A total of 85 (73%) were returned and the anonymised data were entered into SPSS for Windows for analysis. RESULTS: 61% of mothers claimed to visit the pharmacy once a month or more but only 22% of mothers claimed to have had advice from the pharmacist once a month or more. 57% of the mothers are loyal customers to one pharmacy. 87% of the mothers reported that pharmacist's advice was helpful or very helpful. Mothers sought advice from pharmacists for minor childhood ailments but not always in the first instance. Mothers did not feel that the demographic characteristics of the pharmacists affected the advice given but they expressed a personal preference for receiving advice from female pharmacists with children. LIMITATIONS AND RECOMMENDATIONS: This was an exploratory study with a small sample so the results cannot be generalised. Also, convenience sampling is prone to bias. However, this qualitative study presents some preliminary indications about mothers' use of pharmacies for advice on young children's ailments, and suggests aspects that warrant further investigation. CONCLUSION: In this study, mothers of children under five years of age rated pharmacist's advice as helpful. They did not consider that the demographics of the pharmacist would affect the advice they received, but preferred to consult a pharmacist with children. PMID- 15341375 TI - Social exclusion and mental health. PMID- 15341376 TI - A better balance. PMID- 15341377 TI - Not just rice and peas. PMID- 15341378 TI - Behind the scenes. PMID- 15341379 TI - Small changes, big difference. PMID- 15341380 TI - Advocate. PMID- 15341381 TI - Welfare writes. PMID- 15341382 TI - Stigma tsarina. PMID- 15341383 TI - Hindsight. PMID- 15341384 TI - Sharing the knowledge. PMID- 15341385 TI - Same place, same time. PMID- 15341386 TI - It's a different thing. PMID- 15341387 TI - Diatribe. PMID- 15341388 TI - Surviving dual diagnosis. PMID- 15341389 TI - This life. PMID- 15341390 TI - [Ultrasound morphology of prostatic apex: implications for its dissection in prostatectomy]. AB - The prostatic apex can present various morphologies. Myers, on the basis of prostatectomy specimens classified these into two main groups: prostates without an anterior dip (doughnut shaped) and those with an anterior dip (croissant shaped). Surgical duration for prostatectomy known as "apical dissection" must be altered depending on the morphology of the prostatic apex, to prevent incomplete resection of the gland. With transrectal ultrasound performed during transrectal biopsy, the morphology of the apex can be established in each gland. This would help us to plan dissection of the apex in each specific case, avoiding unwanted dissection of the posterior lip of the prostatic apex. PMID- 15341391 TI - [Therapeutic options for patients with localized prostatic carcinoma: our experience with 454 patients]. AB - OBJECTIVES: To evaluate the influence of different therapeutic options on progression-free survival (PFS), overall survival (OS) and specific survival (SS) in a cohort of 454 patients with localized prostatic carcinoma, taking into account different prognostic factors, and to compare our results to those reported in the world literature. MATERIAL AND METHODS: Between 1983 and 2000 we have diagnosed 706 new cases of prostatic carcinoma and 454 were clinically localized tumors. The different therapeutic options employed in our series of patients have been: follow-up (FU) (103 patients); radical prostatectomy (RP) (108 patients); radiotherapy without hormonal blockade (RT) (148 patients); and hormonal blockade (HB) (95 patients). We have determined the PFS, the OS and the SS for each group of patients and compared them in patients with different prognostic factors at the time of diagnosis, including age, PSA levels, Gleason's grading and TNM staging. We have also analysed the influence of the tumor progression on the OS. The mean follow-up time has been 5.6 years (range: 0.1 19.2; median: 5.2). RESULTS: For PFS: the disease progressed in 145 patients (32%) and the PFS at 5 and 10 years has been 77% and 67% for FU; 61% and 50% for RP; 63% and 25% for RT; and 73% and 67% for HB, respectively. The differences between RT and RP were not statistically significant. For the subgroup of patients with PSA levels <10 and Gleason <8 the differences between FU, RP and RT did not reach statistical significance. For OS: 126 patients of our series died (28%) and the OS at 5 and 10 years has been 80% and 61% for FU; 90% and 76% for RP; 85% and 67% for RT; and 64% and 32% for HB, respectively. We have found no significant differences between FU, RP and RT. For SS: 31 patients of our series died of disease (6.8%). The SS at 5 and 10 years has been 100% and 94% for FU; 98% and 98% for RP; 97% and 88% for RT; and 83% and 77% for HB, respectively. We have found no significant differences in the OS between patients with disease progression and without disease progression treated with FU, RP and RT. CONCLUSIONS: Determination of PSA levels has allowed diagnosis of prostatic carcinomas in early stages of disease; however, our results and those reported in the literature cannot define which is the best therapeutic option in these patients. We should offer the patients individualized information both in the phase of early diagnosis and of therapeutic decisions. PMID- 15341392 TI - [Incidence and significance of "atypical acinar proliferation" in transrectal prostatic biopsies]. AB - OBJECTIVE: To determine the incidence of atypical acini in our series, the percentage of prostate cancer and other suspicious or preneoplastic lesions in the successive biopsies and to establish a follow up strategy in these patients. MATERIAL AND METHOD: A total of 117 patients diagnosed with isolated atypical acini or associated with high grade PIN were obtained from our database, 75 (64%) of these were submitted to at least a second biopsy of the gland. The age, PSA, digital rectal examination (DRE), prostate volume, ultrasound nodule and previous pathological diagnosis were compared with the pathology results of the successive biopsies of the gland. RESULTS: Incidence of atypical acini was 4%. A total of 46.7% of patients undergoing a repeat biopsy presented prostate cancer. We did not find significant differences in the clinical variables studied between patients with or without a definite cancer diagnosis. Patients initially diagnosed with acinar atypia with associated high grade PIN presented a higher incidence of cancer in the repeat biopsy than patients with isolated acinar atypia (p=0.007). A total of 94.3% of all neoplasms diagnosed were detected in the 2nd and 3rd biopsy. CONCLUSIONS: Patients with atypical acini are at high risk of having prostate cancer not detected in the first biopsy. They should, therefore, undergo a second biopsy as soon as possible. There is only a very small probability of detecting cancer after the 3rd biopsy. PMID- 15341393 TI - [Analysis of medical/surgical complications of cutaneous ileostomy]. AB - OBJECTIVE: Our objective is to carried out a descriptive study about medical surgical complications and alterations about quality of life in stoma patients secondary to bladder tumour. PATIENTS AND METHOD: The series are composed of 67 patients. The mean age was 70.3 years. We analyse the following variable: early and late medical-surgical complications, the body mass index (BMI) change and the alteration of quality of life. RESULTS: The most frequent early complication is the ischemic necrosis (7%) without posterior repercussion. The late complications in frequency order are: peristomal dermatitis (18%), plane stoma (12%), peristomal hernia (12%), stenosis (9%) and granuloma (6%). The BMI changes don't conditionate a increase in the complications. The psychological adaptation is good in 92%, although the majority affirm a little changes in life-style, but they don't have a repercussion on quality of life. CONCLUSIONS: In this series we estimate a low incidence of complications. The preoperative counsel and the posterior following by the stoma care nurse-surgeon team, play a fundamental role on psychological adaptation after surgery. PMID- 15341394 TI - [Treatment with sildenafil of cold glands syndrome after inflatable penile prosthesis]. AB - OBJECTIVE: To evaluate the effect of sildenafil on the response of glans penis during sexual intercourse in patients with penile prosthesis who show cold glans syndrome symptoms. MATERIAL AND METHODS: Fourteen patients who had undergone three-piece inflatable penile prosthesis implantation were evaluated. In spite of the normal functioning of the device all of them complained of a degree of dissatisfaction during SI due to a lack of engorgement in the glans penis. The patients were advised to have a dose of 100 mg VO sildenafil about 45 minutes before activating the PP and starting SI. All of them followed this scheme at least three times. They ticked questions 7, 8, 13 and 14 of the International Index of Erectile Function (IIEF) questionnaire with/without having had sildenafil. RESULTS: Twelve out of thirteen patients (85.7%) indicated a more pleasant SI on sildenafil. It was related to an increase in penile glans engorgement and sensitivity. The scores obtained in the IIEF questions showed a significant increase on taking sildenafil. The partners also talked of a more comfortable SI that they related to a less painful penetration. Sildenafil related morbidity was not found. CONCLUSIONS: Sildenafil could be used to improve functional results of the penile prosthesis in those patients with CGS and seems to increase satisfaction during SI. PMID- 15341395 TI - [Preservation of the bladder in rectal cancer with prostatic invasion]. AB - Colorectal cancer in the 2nd commonest cancer in Europe. In 5-10% of cases there is infiltration of urological organs. When infiltration affects the bladder or the prostate, anterior pelvic exenteration is the treatment that achieves the largest percentage of tumor free margins and the best 5-year survival. In very select cases of prostatic infiltration, the bladder can be preserved and prostatectomy and abdominoperineal block resection are carried out fulfilling oncological surgical requirements and producing an important improvement in the patient's quality of life. Owing to the very scarce published literature (two articles with three cases) we contribute our experience of 2 patients who received cytoreducing preoperative radiochemotherapy, prostatectomy and block abdominoperineal amputation and intraoperative radiotherapy. We describe the surgical technique used, which is substantially different from standard prostatectomy and requires good coordination between surgeons and urologists. PMID- 15341396 TI - [Prostate cyst at the midline in a patient with a renal transplantation]. AB - Cystic lesions at the midline of the prostate are uncommon and they are considered as congenital disorders. Is generally accepted that active treatment should be instituted only in symptomatic patients and watchful waiting in asymptomatic ones. The most common active treatment is minimally invasive surgery by puncture-aspiration or endoscopic unroofing. We are attaching an inmunosupressed patient because of a renal transplantation in which we provided an active treatment by puncture-aspiration of the cystic lesion in spite of being asymptomatic. PMID- 15341397 TI - [Pseudotumor renal after partial nephrectomy with the use of surgical gelatin sponge]. AB - After renal parenchymal sparing surgery, with the use of surgical gelatin sponge, residual defects may persist on imaging studies at the sites of resection. These "pseudotumors" may lead to confusion as to whether a lesion was removed or has recurred. These lesions usually resolve within a year. We report a case of pseudotumor renal parenchymal sparing surgery in a 69 year old man, with resolution of the lesion 8 months after surgery. PMID- 15341398 TI - [Metastasis in renal graft of primary renal adenocarcinoma]. AB - Acquired cystic renal disease (A.C.R.D.) is a risk factor for the development of renal carcinoma in the patient's own kidney after renal transplant (R.T.), development of a primitive tumor in the renal graft is infrequent, but the presence of metastasis of an epithelial tumor is very rare. This is the second case reported in the literature of metastasis of an epithelial tumor in the renal graft and the first case described of renal cell carcinoma metastasis (R.C.C.) in the renal graft. This paper describes the case of a patient with a normally functioning renal transplant and A.C.R.D. who develops RCC in his own kidney, with metastasis in the renal graft. PMID- 15341399 TI - [Kidney pelvis hemangioma as a cause of hematuria]. AB - Kidney hemangiomas are congenital vascular anomalies with an unknown ethiology. They are considered as benign tumours with a poor clinic expression (hematuria or ureteric colic, which is associated to the drop of clots in the urinary tract). OBJECTIVE: To expound a case of unilateral severe hematuria in a young woman with a restless past medical history, due to a pelvic left kidney hemangioma. MATERIALS AND METHODS: Clinic case of a sixteen years old woman, no relevant past medical history, presented with an anaemizing gross hematuria and passing vermiform clots. As diagnostic techniques we used ultrasonography, excretory urography, cystourethroscopy, CT, angiography and nuclear imaging. Surgery (flank nephectomy) was needed for treatment. RESULTS: Pathology report: submucosal vascular proliferative process in kidney pelvis composed by blood vessels, suggesting haemangioma. PMID- 15341400 TI - [Iatrogenic prostatorectal fistula. Use of gracilis muscle pedicled flap for its reparation. Presentation of two cases]. AB - We present two cases in which during retropubic prostatectomy for benign prostatic disease a prostatorectal fistula ocurred. We describe its reparation using a pedicled flap of gracilis muscle. We also present cystographic and opaque enema images which shows the before and after of this surgery. Patients had good outcome without incontinence nor problems related to muscle desinsertion surgery. PMID- 15341401 TI - [Gastric metastasis from renal cell carcinoma. Pathogenical hypothesis and literature revision]. AB - Secondary gastric tumours are very uncommon clinical entities and even more so when the site for the primary tumour is the kidney. Only 11 cases of life diagnosis have been described up to now. Contribution of one case report: a female patient presenting with upper GI tract haemorrhage secondary to gastric metastasis from renal cell carcinoma four years after radical nephrectomy. A literature review is made on the cases described up to date in living patients and a pathogenic hypothesis established based on the theoretical dissemination routes for the cases of gastric location of the metastasis. PMID- 15341402 TI - [Low grade collecting duct carcinoma of the kidney. Presentation of one case and literature revision]. AB - Collecting duct renal cell carcinoma is an uncommon variant of renal carcinoma. Typically its behaviour is more aggressive than other forms of renal carcinoma and usually it is diagnosed at advanced stages. A 57-year-old man visited our hospital by right lumbar pain. Abdominal CT showed a enhanced mass on the right kidney. A right partial nephrectomy was done. Histological examination of the surgical specimen showed a low grade collecting duct carcinoma of the kidney. This is a new entity, with unknown behaviour but seems to be less aggressive than classical collecting duct carcinoma of the kidney. The clinical, radiological, pathological and immunohistochemical characteristics of this tumor are reported, and the literature is reviewed. PMID- 15341405 TI - More cost drivers. PMID- 15341406 TI - Paying for ED call. PMID- 15341407 TI - Interpersonal skills undervalued. PMID- 15341408 TI - Capital purchasing problems. PMID- 15341409 TI - Technology. The standards wait. PMID- 15341410 TI - Technology. Incentives, not mandates. PMID- 15341411 TI - Facilities. My generation. PMID- 15341412 TI - Payment. Insurance bypass. PMID- 15341413 TI - HIPAA. Security gaps. PMID- 15341414 TI - Access & coverage. Will work for care. PMID- 15341415 TI - Data page. Collecting race & ethnicity data. PMID- 15341416 TI - There's no 'I' in 'team'. AB - McKesson Corp. CEO John Hammergren shies away from the spotlight. He credits a strong staff and customer focus for the company's success. PMID- 15341417 TI - What if... [the consequences of innovation]. AB - Innovations that could turn health care upside down are already here, in use or in development. These technological or clinical "wild cards" so resemble science fiction that they are easily dismissed--for example, brain implants to help the disabled control remote appliances or human arteries grown from scratch. Ignore them at your own risk. PMID- 15341418 TI - HSAs: are they really the next big thing? AB - Advocates say health savings accounts will kick-start the era of consumer-driven health care and bring costs under control. Critics say they are perks for the well-to-do and will increase the number of uninsured Americans. What's clear is that HSAs have the potential to dramatically change how consumers use and pay for your services. PMID- 15341419 TI - Circle of Life Awards 2004. AB - This year's winners of the American Hospital Association's Circle of Life Awards exemplify the expanding nature of palliative care. They are finding creative ways to offer solace and pain relief not just at the end of life, but also earlier in the course of a life-threatening illness. And they're looking beyond the traditional hospice borders. PMID- 15341420 TI - Critical access hospitals. AB - A variety of factors are coming together that will ignite an explosion of conversions to critical access hospital status. What hospitals are eligible to become CAHs, what are the benefits to converting and what should hospital leaders consider before taking the plunge? PMID- 15341421 TI - Low-income Americans face serious gaps in specialty care for physicians, mental health and dental care. PMID- 15341422 TI - Leapfrog standards for high-risk surgeries could save 8,000 lives, study finds. PMID- 15341423 TI - Growing number of adults use complementary and alternative medicine. PMID- 15341424 TI - The quest for quality. AB - By adopting six Institute of Medicine goals, the Quest for Quality Prize has put the focus on improving systems of care. PMID- 15341425 TI - Objects look different sizes in the right and left eyes. AB - Coren and Porac (1976) reported that objects looked larger in the right eye of right-eye dominant subjects and in the left eye of left-eye dominant subjects. This paper attempts to repeat that finding. Two circles of same or different size were presented haploscopically in a binocular three-field tachistoscope, to right or left visual half-field and to the upper or lower visual field, one to the right eye and one to the left. A total of 43 subjects reported which of the two circles was the larger, each subject carrying out 120 trials of the experiment. Overall subjects reported that the stimulus to the left eye was significantly larger than that presented to the right eye. There was no association with eye dominance, and therefore the Coren and Porac finding could not be repeated. There was however a very significant association with handedness, left-handed subjects tending to report that the stimulus in the right eye looked larger, and right handed subjects reporting that the stimulus in the left eye looked larger. PMID- 15341426 TI - Biobehavioural correlates of hand preference in free-ranging female primates. AB - In this research we examined biological and behavioural correlates of handedness in a subject cohort of 41 free-ranging young female rhesus macaques (Macaca mulatta). Specifically, we examined relationships between handedness and cerebrospinal fluid (CSF) concentrations of the monoamine metabolites 5 hydroxyindoleacetic acid (5-HIAA) and homovanillic acid (HVA), plasma concentrations of the hormones cortisol and adrenocorticotropin (ACTH), prolactin, and multiple indices of social behaviour, including proximity to other animals, grooming, submission, and aggression. Handedness was determined through systematic observation of animals reaching for food in their unrestricted home environment. We found a population-level bias for left-hand use in this cohort of young females. The frequency of right versus left hand use was positively correlated with CSF 5-HIAA, plasma cortisol concentrations, the frequency of submissive behaviour, and with the frequency of bouts in which animals received low-level aggression. The positive correlation between right versus left hand use, submissive behaviour, and received aggression found here in females contrasts with the negative correlation among these same variables that we have previously reported in rhesus males. We conclude that these results may be explicable in terms of sex-based differences in rhesus life-history patterns, and that the influence of the serotonergic system on patterns of male aggression, social behaviour, and handedness, and the associations between handedness and social behaviour found previously among males may not be generalised to female rhesus macaques. PMID- 15341427 TI - Effects of culture, sex, and age on the distribution of handedness: an evaluation of the sensitivity of three measures of handedness. AB - The present study examined the effects of culture, age, and sex on three measures of handedness: writing hand, Annett's primary handedness items, and a measure based on hand preference for 11 activities. Using data from a large international study, the relationship between the culture in which participants learned to write (as defined by Hofstede's cultural dimensions) and adult handedness was examined. Participants who learned to write in formal cultures were less likely to be classified as left-handed than those who learned in less formal cultures. Older participants and those who learned to write in formal cultures were more likely to be classified as left-handed by the Annett and 11 item measures than by the writing hand classification. Across measures females were less likely to be classified as left-handed than males. Handedness for writing was found to be more sensitive to cultural influences than the other measures. These results suggest that some measures of handedness may be more sensitive to specific handedness aetiologies than others. PMID- 15341428 TI - Aiming in adults: sex and laterality effects. AB - The purpose of the study was to twofold: to investigate gender-related differences in the asymmetry of aiming with the preferred and non-preferred hand in right-handed adults, and to examine the effect of the spatial requirements of the task on these asymmetries. The hypothesis was that if cognitive functions are more asymmetrically localised in men than in women, one should observe greater left-right differences on some variables in men than in women. Eleven men and eleven women were required to aim fast and accurately at one of three possible targets under a choice reaction time protocol. Performance and kinematics data were analysed. Results revealed an effect of target location on the left hand advantage in reaction time, and gender-related effects on movement time, accuracy, and on the velocity profiles. Overall, women performed more slowly and accurately than men. This gender-related effect could not be accounted for by differential strategies with regard to speed or accuracy, lending support to the idea that differences exist in the neural mechanisms of movement control between the two genders. Finally, although the results show a hand effect on terminal accuracy in men only, they do not support the hypothesis of a greater asymmetry of movement control in men. PMID- 15341429 TI - The role of the cerebral hemispheres in specific versus abstract priming. AB - The present study examined specific and abstract priming for pictures in unilateral brain-damaged patients using a picture fragment identification task. Participants were 10 posterior right hemisphere (RH) damaged patients, 11 posterior left hemisphere (LH) damaged patients, and 30 healthy normal controls. Results reveal that RH patients perform specific priming tasks significantly worse than healthy participants (p < .005). There was a trend (p < .1) towards worse abstract priming in LH patients when compared to healthy participants. These results are additional evidence that specific priming depends on normal functioning of the posterior right cerebral hemisphere, while abstract priming depends on normal functioning of the posterior left cerebral hemisphere, and extend this finding from verbal to non-verbal stimuli. PMID- 15341430 TI - Hand preference in patients with allergy, juvenile cancer, and schizophrenia. AB - Patients with allergy (n = 50), juvenile cancer (n = 50), and schizophrenia (n = 37), and nonpatient controls (n = 200) were asked to indicate their hand preferences on a 7-point scale (1 = left always, 7 = right always) for the 32 item Waterloo Handedness Questionnaire. Except for patients with schizophrenia, who exhibited a higher incidence of mixed hand preference, groups showed a clear rightward bias. Further analyses suggested that mixed hand preference in schizophrenia was evident more often for unskilled than skilled hand activities. PMID- 15341431 TI - Handedness and spatial ability: differential patterns of relationships. AB - The purpose of this exploratory study was to add to our understanding of hemispheric specialisation by examining the relationships between functional laterality (as measured by handedness, eyedness, earedness, and footedness) and spatial tests. A total of 55 males and 170 females were tested for laterality preferences and spatial test performance using a range of six spatial tests. There was a statistically significant relationship between each of the laterality measures employed, with handedness and footedness demonstrating the most robust association. Multiple regression analyses highlighted that once potential confounds were statistically controlled (age, sex, and other types of functional laterality), 3-D rotation/visualisation and speeded visual exploration task performance showed small, but significant, relations with a tendency towards left handedness over right-handedness in total and secondary (two-handed) scale scores. Flexibility of closure performance was associated with a tendency towards left-handedness in the total and one-handed scale scores. Spatial location memory was the lone spatial ability relating with a tendency towards right-handedness. Overall, these findings offer new insights into the interplay of handedness with alternative measures of laterality as well as with spatial abilities. PMID- 15341432 TI - In the terminally ill, a wish to die is a manifestation of depression and should be treated accordingly. PMID- 15341433 TI - The wish to die in the terminally ill population is indicative of depression and should be treated accordingly. PMID- 15341434 TI - Proton therapy: expanding clinical indications. PMID- 15341435 TI - First spinal axis segment irradiation with spot-scanning proton beam delivered in the treatment of a lumbar primitive neuroectodermal tumour. Case report and review of the literature. AB - Primary intraspinal primitive neuroectodermal tumour (PNET) is a rare tumour entity. The optimal therapeutic management is unclear but, in general, this tumour is treated with surgery followed by radiotherapy and chemotherapy. Proton beam radiation therapy (PT) offers superior dose distributional qualities compared with X- or gamma rays, as the dose deposition occurs in a modulated narrow zone called the Bragg peak. As a result, organs at risk are optimally speared. Here, we present a patient treated with the first spinal axis segment irradiation using spot-scanning PT with a single field, combined with conventional cranio-spinal axis radiotherapy after surgery and chemotherapy, and an extensive review of the literature outlining the clinical features and treatment modality of spinal PNET. PMID- 15341436 TI - Prognostic factors in brain metastases: can we determine patients who do not benefit from whole-brain radiotherapy? AB - AIMS: Whole-brain radiotherapy (WBRT) is a standard treatment recommendation for patients with brain metastases. The goal of treatment is symptom control, which in the short run can be often achieved by steroids. Patients with a short life expectancy may not derive benefit from the addition of radiation. The ability to identify this group would aid the decision of whether to recommend WBRT. MATERIALS AND METHODS: Data on all patients referred for WBRT to palliative radiotherapy teams at two comprehensive cancer centres were prospectively collected over a 2-year period. The most frequent radiation dose and fractionation was 2000 cGy in 5 fractions over 1 week. Multivariate logistic regression analysis using forward stepwise selection process was used to develop a prediction model for early death (before 8 weeks). The variables considered were sex, performance status, primary disease, weight loss, age, time from primary diagnosis to brain metastases diagnosis, number of metastatic sites and Radiation Therapy Oncology Group (RTOG) recursive partitioning analysis (RPA) status. RESULTS: Two hundred and seventy-five patients with brain metastases were assessed. The median follow-up was 19 weeks, and estimated median overall survival was 5.3 months. Multivariate regression revealed Eastern Cooperative Oncology Group (ECOG) performance status and number of metastatic sites to be significant predictors of early death. The odds ratios were 2.38 (95% confidence interval [CI] 1.77-3.19) and 1.39 (95% CI 1.07-1.81), respectively. Sixty-eight per cent of patients could be correctly classified; however, 55% would have been incorrectly predicted to die early. CONCLUSIONS: Poor performance status and number of metastatic sites are useful predictors of early death. A regression model was highly predictive overall, but this was primarily due to a high negative predictive value of 86%. The ability to predict patients who would die early (positive predictive value) was only 45%. Thus, despite the understanding that we are over-treating a subset of patients, further research is required to identify patients who do not require radiotherapy. PMID- 15341437 TI - Defining the impact and contribution of steroids in patients receiving whole brain irradiation for cerebral metastases. AB - AIMS: To assess the impact of steroid therapy in the management of brain metastases by review of the published literature on whole-brain radiotherapy (WBRT) for multiple cerebral metastases. MATERIALS AND METHODS: Twenty-one full manuscripts of published randomised controlled trials (RCTs) involving WBRT for multiple cerebral metastases were identified from a literature search and included studies from 1971--2003. Details on the use and type of steroid, timing of steroids relative to radiotherapy, response assessment and contribution of steroids to overall outcome were extracted. RESULTS: Eighteen out of 21 trials commented on steroid use. All studies used overall survival as an assessment of outcome; 13 studies assessed neurological response and 10 used imaging assessment. Ten studies acknowledged a contribution from steroids in the response assessment or analysis, but only one study gave a fixed steroid dose to control for its palliative effect. CONCLUSIONS: Reporting of steroid use in published RCTs assessing treatment for patients with brain metastases is non-uniform and not sufficiently detailed. As such, it is difficult to assess the additional benefit of WBRT. A standardised approach to the incorporation of steroid effect in assessment of WBRT would be advantageous to determine symptom response or durability of response. Side-effects of steroid use and the ability to taper off steroids after treatment intervention are also important outcomes of interest. PMID- 15341438 TI - Brain metastases from breast cancer: identification of a high-risk group. AB - AIMS: Brain metastases from breast cancer are an uncommon initial presentation of metastatic breast cancer, but brain metastases commonly occur later in women's metastatic illness. The aims of this study were to document the type, frequency, and temporal occurrence of brain metastases from breast cancer as well as the survival of women with such metastases, and to attempt to identify a subgroup of women at high risk of brain metastases who may benefit from pre-emptive medical intervention. MATERIALS AND METHODS: The radiological reports of all women presenting with metastases aged under 70 years who had subsequently died were examined. The type, frequency, temporal occurrence and survival with brain metastases were documented. Correlations were sought between the frequency of brain metastases and age at metastatic presentation, tumour grade, histological type and oestrogen receptor (ER) status. RESULTS: Of 219 patients who had died with metastatic disease and who were under 70 years of age at metastatic presentation, 49 (22%) developed brain metastases. The development of brain metastases was related to young age (P = 0.0002), with 43% of women under 40 years developing brain metastases. Brain metastases were more common in women whose tumours were ER negative (38%) compared with women with ER-positive disease (14%) (P = 0.0003). By combining age and ER status, it is possible to identify a group of women (age under 50 years and ER negative) with a 53% risk of developing brain metastases. This group included many women who had chemotherapy for visceral metastases, and 68% had either stable disease or disease response at other sites at the time of brain metastases presentation. CONCLUSION: It is possible to identify a subgroup of women with metastatic breast cancer at high risk of brain metastases who may benefit from pre-emptive medical intervention, such as screening or prophylactic treatment. PMID- 15341439 TI - Treatment results of endometrial carcinoma with positive peritoneal washing, adnexal involvement and serosal involvement. AB - AIMS: To review the treatment results of patients with endometrial carcinoma having positive peritoneal washing (PPW), adnexal involvement, uterine serosal involvement, or all three. MATERIALS AND METHODS: The treatment records of patients who had undergone primary surgery for endometrial cancer without distant metastasis during 1990--2001 at the Department of Clinical Oncology, Tuen Mun Hospital, Hong Kong, were reviewed. Thirty-five patients were found to have involvement of positive PPW, adnexal involvement, uterine serosal involvement, or all three. Seven (20%) of them had gross or microscopic lymph-node metastasis. Thirty-three (94.3%) patients received adjuvant radiotherapy (28 whole-pelvic irradiation [WPI]; five abdominal radiotherapy [WART]). Two patients with solitary ovarian metastasis received chemotherapy, and one with isolated PPW also received adjuvant hormonal therapy. The median follow-up was 50.4 months (range 2.4-151.2 months). Multivariate analysis was carried out using the Cox regression proportional hazards model. RESULTS: Among the 28 patients with clinical or pathological node-negative disease (International Federation of Gynecology and Obstetrics [FIGO] stage IIIA), only two patients with solitary ovarian metastases developed recurrence. The 5-year actuarial disease-free survival (DFS) rates for the whole group and patients without lymph-node involvement were 77.9% and 91.7%, respectively. Five out of the seven patients with lymph-node involvement developed recurrences. Univariate analysis showed that lymph-node involvement (P < 0.0001) and high-grade disease (P = 0.011) were the significant poor prognostic factors. Multivariate analysis showed that lymph-node involvement was the only significant poor prognostic factor to predict poor 5-year DFS (P = 0.0001). Only one patient (3.7%) who had received WART developed grade 4 toxicity. CONCLUSIONS: This study showed that good treatment results could be obtained from patients with stage IIIA endometrial carcinoma without clinical or pathological lymph-node involvement after adjuvant radiotherapy, with acceptable late side-effects. The relative prognostic importance of individual IIIA involvement and the optimal adjuvant treatment remain to be determined. PMID- 15341440 TI - Cellular immunotherapy for cancer: current concepts and clinical perspectives scientific basis and approaches for therapeutic cancer vaccines. Part I. PMID- 15341441 TI - Use of cytological specimens for p53 gene alteration detection in oral squamous cell carcinoma risk patients. AB - AIMS: Recurrence and multifocal nature are two important characteristics of oral squamous cell carcinoma. Leukoplakia is the most frequent pre-cancerous oral lesion and, in most cases, it is not possible to predict malignant capacity. The objective of this study is to identify p53 alterations in cells taken from the oral cavity of at-risk patients. MATERIALS AND METHODS: The following samples were collected from 34 patients with oral leukoplakia with and without previous carcinoma: oral rinse, a brush swabbed over the lesions and hair roots. Mutational analysis of the p53 gene was performed by single-strand conformation polymorphisms and confirmed by DNA sequencing. RESULTS: We detected 11 mutations in p53 gene in oral cytological specimens. These alterations were observed only in brush cytology samples in patients without previous carcinoma, and in both samples (rinse and brush) in patients with previous carcinoma. Three of these patients had disease recurrence. CONCLUSION: This non-invasive technique may be useful in the follow-up of at-risk patients, and introduces new possibilities to analyse molecular markers before malignant lesions are clinically apparent. PMID- 15341442 TI - A patient with Rothmund-Thomson syndrome and tongue cancer--experience of radiation toxicity. AB - We describe a male patient with Rothmund-Thomson syndrome (RTS) given postoperative radiotherapy for squamous carcinoma of the tongue. This was well tolerated. This is only the second reported case of oral cancer and radiotherapy in RTS. PMID- 15341443 TI - Learning in oncology: lessons from the 20th century, learner-centred education for the 21st century. Part I. PMID- 15341444 TI - Bisphosphonate use in metastatic breast cancer: an audit assessing implementation of the British Association of Surgical Oncologists' guidelines. PMID- 15341445 TI - Contemporary non-small cell lung cancer practice in the United Kingdom. PMID- 15341446 TI - A rare presentation of primary fallopian tube carcinoma. PMID- 15341447 TI - Biomedical applications of high-energy ion beams. PMID- 15341448 TI - ECF in gastric cancer. PMID- 15341449 TI - Fluid resuscitation with albumin or saline in the intensive care unit did not affect 28-day mortality rates. PMID- 15341450 TI - An antibiotic regimen for 8 days was as effective as one for 15 days in ventilator-associated pneumonia. PMID- 15341451 TI - Immediate carotid endarterectomy reduced nonperioperative stroke in severe asymptomatic carotid artery stenosis. PMID- 15341452 TI - Cholesterol lowering with simvastatin reduced stroke in patients with, or at risk for, vascular disease. PMID- 15341453 TI - High-dose atorvastatin was superior to standard-dose pravastatin for reducing death or major cardiovascular events in acute coronary syndromes. PMID- 15341454 TI - Percutaneous transluminal coronary angioplasty improved long-term angina status more than medical therapy in coronary artery disease. PMID- 15341455 TI - Rapid measurement of B-type natriuretic peptides reduced time to discharge and treatment costs in patients with acute dyspnea. PMID- 15341456 TI - Sildenafil was safe and improved erectile function and quality of life in men with moderately severe congestive heart failure. PMID- 15341457 TI - Doubling the dose of inhaled corticosteroids during asthma deterioration did not improve asthma control. PMID- 15341458 TI - Memantine was better than placebo in Alzheimer disease already being treated with donepezil. PMID- 15341459 TI - Review: donepezil improves cognitive and functional outcomes in vascular cognitive impairment. PMID- 15341460 TI - Postexposure prophylaxis with oseltamivir reduced influenza transmission in households. PMID- 15341461 TI - Review: Helicobacter pylori eradication therapy is mildly effective for reducing symptoms in nonulcer dyspepsia. PMID- 15341462 TI - Etanercept plus methotrexate reduced symptoms and disease activity in adult-onset rheumatoid arthritis. PMID- 15341463 TI - Review: thoracoscopic pleurodesis with talc may be the optimal technique in patients with malignant pleural effusions. PMID- 15341464 TI - Review: tegaserod prompts global relief of symptoms in irritable bowel syndrome. PMID- 15341465 TI - Review: leukotriene-receptor antagonists are less effective than intranasal corticosteroids for allergic rhinitis. PMID- 15341466 TI - Review: skin and in vitro tests for allergic rhinitis vary widely in usefulness. PMID- 15341467 TI - Review: vaginal signs and symptoms perform poorly in diagnosing vaginal candidiasis, bacterial vaginosis, and vaginal trichomoniasis. PMID- 15341468 TI - Chest radiographs and BNP levels provided complementary information beyond clinical findings for diagnosing heart failure. PMID- 15341469 TI - Review: 4 clinical signs are precise and accurate for predicting poor outcome in postcardiac arrest coma. PMID- 15341470 TI - Elevated plasma natriuretic peptide levels were associated with cardiovascular events. PMID- 15341472 TI - Review: individualized written action plans based on peak expiratory flow improve asthma health outcomes. PMID- 15341471 TI - C-reactive protein was a moderate predictor of coronary heart disease. PMID- 15341473 TI - The BODE index predicted death in chronic obstructive pulmonary disease. PMID- 15341474 TI - Contribution of the Cochrane Library to the evidence-based journals. PMID- 15341476 TI - The first symptom of hyperkalemia is death. PMID- 15341477 TI - Virulence regulation and quorum sensing in staphylococcal infections: competitive AgrC antagonists as quorum sensing inhibitors. PMID- 15341478 TI - Potent and orally bioavailable non-peptide antagonists at the human bradykinin B(1) receptor based on a 2-alkylamino-5-sulfamoylbenzamide core. AB - The bradykinin B(1) receptor is rapidly induced after inflammation or tissue trauma and appears to play an important role in the maintenance of hyperalgesia in inflammatory conditions. Here, we describe the optimization process to identify novel, potent non-peptide human B(1) receptor antagonists based on a 2 alkylamino-5-sulfamoylbenzamide core. Optimized derivatives are selective, functional B(1) antagonists with low nanomolar affinity and exhibit oral bioavailability in animals. PMID- 15341479 TI - Discovery of novel heteroarylazoles that are metabotropic glutamate subtype 5 receptor antagonists with anxiolytic activity. AB - The highly potent, selective, and brain-penetrant metabotropic glutamate subtype 5 (mGlu5) receptor antagonists 3-(5-pyridin-2-yl-2H-tetrazol-2-yl)benzonitrile (47) and 3-fluoro-5-(5-pyridin-2-yl-2H-tetrazol-2-yl)benzonitrile (48) are reported. Compound 47 is active in the rat fear-potentiated startle (FPS) model of anxiety with ED(50) = 5.4 mg/kg (po) when dosed acutely. In this model the anxiolytic effects of 47 rapidly tolerate on repeated dosing. PMID- 15341480 TI - In vitro cytotoxic activities of 2-alkyl-4,6-diheteroalkyl-1,3,5-triazines: new molecules in anticancer research. AB - The cytotoxic activities of new 2-alkyl-4,6-dihetero(N,O)alkyl-1,3,5-triazines toward selected tumor cell lines have been evaluated, and for the first time, the potential of 2-alkyl-4,6-dialkoxy-1,3,5-triazines has been shown. PMID- 15341481 TI - Fuzzy pharmacophore models from molecular alignments for correlation-vector-based virtual screening. AB - A pharmacophore-based approach for compiling focused screening libraries is presented. It integrates information from three-dimensional molecular alignments into correlation vector-based database screening. The pharmacophore model is represented by a number of spheres of Gaussian-distributed feature densities. Different degrees of "fuzziness" can be introduced to influence the model's resolution. Transformation of this pharmacophore representation into a correlation vector results in a vector of feature probabilities which can be utilized for rapid virtual screening of compound databases or virtual libraries. The approach was validated by retrospective screening for cyclooxygenase 2 (COX 2) and thrombin ligands. A variety of models with different degrees of fuzziness were calculated and tested for both classes of molecules. Best performance was obtained with pharmacophore models reflecting an intermediate degree of fuzziness, yielding an enrichment factor of up to 39 for the first 1% of the ranked database. Appropriately weighted fuzzy pharmacophore models performed better in retrospective screening than similarity searching using only a single query molecule. The new pharmacophore method was shown to complement existing approaches. PMID- 15341482 TI - Structural basis for the potent antisickling effect of a novel class of five membered heterocyclic aldehydic compounds. AB - Naturally occurring five-membered heterocyclic aldehydes, including 5 hydroxymethyl-2-furfural, increase the oxygen affinity of hemoglobin (Hb) and strongly inhibit the sickling of homozygous sickle red blood (SS) cells. X-ray studies of Hb complexed with these compounds indicate that they form Schiff base adducts in a symmetrical fashion with the N-terminal alphaVal1 nitrogens of Hb. Interestingly, two cocrystal types were isolated during crystallization experiments with deoxygenated Hb (deoxyHb): one crystal type was composed of the low-affinity or tense (T) state Hb quaternary structure; the other crystal type was composed of high-affinity or relaxed state Hb (with a R2 quaternary structure). The R2 crystal appears to be formed as a result of the aldehydes binding to fully or partially ligated Hb in the deoxyHb solution. Repeated attempts to crystallize the compounds with liganded Hb failed, except on rare occasions when very few R state crystals were obtained. Oxygen equilibrium, high performance liquid chromatography (HPLC), antisickling, and X-ray studies suggest that the examined heterocyclic aldehydes may be acting to prevent polymerization of sickle hemoglobin (HbS) by binding to and stabilizing liganded Hb in the form of R2 and/or various relaxed state Hbs, as well as binding to and destabilizing unliganded T state Hb. The proposed mechanism may provide a general model for the antisickling effects of aldehyde containing small molecules that bind to N terminal alphaVal1 nitrogens of Hb. The examined compounds also represent a new class of potentially therapeutic agents for treating sickle cell disease (SCD). PMID- 15341483 TI - Indolebutylamines as selective 5-HT(1A) agonists. AB - A series of new 1-[4-(indol-3-yl)butyl]-4-arylpiperazines was prepared to identify highly selective and potent 5-HT(1A) agonists as potential pharmacological tools in studies of mood disorders. The combination of structural elements (indole-alkyl-amine and aryl-piperazine) known to introduce 5-HT(1A) receptor affinity and the proper selection of substituents (R on the indole moiety and R' on the aryl moiety) led to compounds with high receptor specificity and affinity. In particular, the introduction of the methyl ether or the unsubstituted carboxamide as substituents in position 5 of the indole (R) guaranteed serotonergic 5-HT(1A) affinity compared to the unsubstituted analogue. Para-substituted arylpiperazines (R') decreased dopaminergic D(2) binding and increased selectivity for the 5-HT(1A) receptor. Agonistic 5-HT(1A) receptor activity was confirmed in vivo in the ultrasonic vocalization test, and the results suggest that the introduction of the carboxamide residue leads to better bioavailability than the corresponding methyl ether. 3-[4-[4-(4 Carbamoylphenyl)piperazin-1-yl]butyl]-1H-indole-5-carboxamide 54 was identified as a highly selective 5-HT(1A) receptor agonist [GTPgammaS, ED(50) = 4.7 nM] with nanomolar 5-HT(1A) affinity [IC(50) = 0.9 nM] and selectivity [D(2), IC(50) > 850 nM]. 3-[4-[4-(4-Methoxyphenyl)piperazin-1-yl]butyl]-1H-indole-5-carboxamide 45 is one of the most potent and selective 5-HT(1A) agonists known [5-HT(1A), IC(50) = 0.09 nM; D(2), IC(50) = 140 nM]. PMID- 15341484 TI - Synthesis and structure--activity relationship in a class of indolebutylpiperazines as dual 5-HT(1A) receptor agonists and serotonin reuptake inhibitors. AB - Systematic structural modifications of indolealkylphenylpiperazines led to improved selectivity and affinity within this class of 5-HT(1A) receptor agonists. Introduction of electron-withdrawing groups in position 5 on the indole raises serotonin transporter affinity, and the cyano group proved to be the best substituent here. 5-Fluoro and 5-cyano substituted indoles show comparable results in in vitro and in vivo tests, and bioisosterism between these substituents was supported by calculation of the molecular electrostatic potentials and dipole moments. Compounds showing promising in vitro data were further examined in ex vivo (p-chloroamphetamine assay) and in vivo (ultrasonic vocalization) tests. Optimization of the arylpiperazine moiety indicated that the 5-benzofuranyl-2-carboxamide was best suited to increase 5-HT transporter and 5 HT(1A) receptor affinity and to suppress D(2) receptor binding. 5-[4-[4-(5-Cyano 3-indolyl)butyl]-1-piperazinyl]benzofuran-2-carboxamide 29 (vilazodone, EMD 68843) was identified as a highly selective 5-HT(1A) receptor agonist [GTPgammaS, ED(50) = 1.1 nM] with subnanomolar 5-HT(1A) affinity [IC(50) = 0.2 nM] and as a subnanomolar 5-HT reuptake inhibitor [RUI = 0.5 nM] showing a great selectivity to other GPCRs (e.g., D(2), IC(50) = 666 nM). PMID- 15341485 TI - A prodrug approach toward the development of water soluble fluoroquinolones and structure--activity relationships of quinoline-3-carboxylic acids. AB - A fluoroquinolone prodrug, PA2808, was prepared and shown to convert to the highly active parent drug PA2789. In vitro and in vivo activation of PA2808 by alkaline phosphatase was demonstrated using disk diffusion and rat lung infection models. The water solubility of PA2808 showed a marked increase compared to PA2789 over a pH range suitable for aerosol drug delivery. A total of 48 analogues based on PA2789 were prepared and screened against a panel of Gram positive and Gram-negative pathogens. Incorporating a cyclopropane-fused pyrrolidine (amine) at C-7 resulted in some of the most active analogues. PMID- 15341486 TI - Design and synthesis of a nitrogen mustard derivative stabilized by apo neocarzinostatin. AB - Neocarzinostatin (NCS) is an antitumor antibiotic comprising a 1:1 protein chromophore complex and exhibits cytotoxic action through DNA cleavage via H abstraction. Cytotoxic activity resides with the chromophore 1 alone, while the protein (apoNCS) protects and transports labile 1. The naphthoate portion (2) of NCS chromophore (1) is important for binding to apoNCS and DNA intercalation. In this paper we describe our attempts to use apoNCS to improve the hydrolytic stability of novel bifunctional DNA alkylating agents. The nitrogen mustards, melphalan and chlorambucil, were both conjugated to 2, and the biological activities of these conjugates were assessed. Chlorambucil did not benefit from conjugation. The melphalan conjugate (6) formed covalent DNA adducts at guanine bases and exhibited greater in vitro cytotoxic activity than unmodified melphalan. Fluorescence and NMR spectroscopy showed that 6 binds to apoNCS. Binding to apoNCS-protected 6 reduced the extent of hydrolysis of the conjugate. This novel approach demonstrates for the first time that an enediyne apo-protein can be used to improve the stability of substances that are of potential interest in cancer chemotherapy. PMID- 15341487 TI - N-Phenyl-4-pyrazolo[1,5-b]pyridazin-3-ylpyrimidin-2-amines as potent and selective inhibitors of glycogen synthase kinase 3 with good cellular efficacy. AB - Glycogen synthase kinase 3 regulates glycogen synthase, the rate-determining enzyme for glycogen synthesis. Liver and muscle glycogen synthesis is defective in type 2 diabetics, resulting in elevated plasma glucose levels. Inhibition of GSK-3 could potentially be an effective method to control plasma glucose levels in type 2 diabetics. Structure-activity studies on a N-phenyl-4-pyrazolo[1,5 b]pyridazin-3-ylpyrimidin-2-amine series have led to the identification of potent and selective compounds with good cellular efficacy. Molecular modeling studies have given insights into the mode of binding of these inhibitors. Since the initial leads were also potent inhibitors of CDK-2/CDK-4, an extensive SAR was performed at various positions of the pyrazolo[1,5-b]pyridazin core to afford potent GSK-3 inhibitors that were highly selective over CDK-2. In addition, these inhibitors also exhibited very good cell efficacy and functional response. A representative example was shown to have good oral exposure levels, extending their utility in an in vivo setting. These inhibitors provide a viable lead series in the discovery of new therapies for the treatment of type 2 diabetes. PMID- 15341488 TI - CORES: an automated method for generating three-dimensional models of protein/ligand complexes. AB - We describe a new, automated method for building 3D models of small-molecule ligands complexed with proteins. Modeling templates are constructed from frameworks (i.e., ring systems and linkers) of ligands extracted from 3D structures of ligands complexed with proteins that are structurally related to the target protein. These templates are typically substructures of the target ligand and are used to build models that constrain the ligand's conformation and binding orientation in the active site of the target protein. The practical utility of the method is shown by demonstrating that most ligands containing related frameworks bind protein kinases in the same orientation. Moreover, models for 15 of 19 cdk2/ligand complexes in the protein data bank built using our method deviate from the X-ray structure by less than 2 A (rms). Finally, we show that over 70% of small-molecule protein kinase inhibitors published in J. Med. Chem. since 1993 can be modeled using a template extracted from a 3D protein kinase structure in the protein data bank. PMID- 15341489 TI - Design, synthesis, and biological evaluation of 1,2,3,7-tetrahydro-6h-purin-6-one and 3,7-dihydro-1h-purine-2,6-dione derivatives as corticotropin-releasing factor(1) receptor antagonists. AB - A growing body of evidence suggests that CRF(1) receptor antagonism offers considerable therapeutic potential in the treatment of diseases resulting from elevated levels of CRF, such as anxiety and depression. A series of novel 1,2,3,7 tetrahydro-6H-purin-6-one and 3,7-dihydro-1H-purine-2,6-dione derivatives was synthesized and evaluated as corticotropin releasing factor-1 (CRF(1)) receptor antagonists. Compounds within this series, represented by compound 12d (IC(50) = 5.4 nM), were found to be highly potent CRF(1) receptor antagonists. In addition, compounds 12d and 12j were determined to be selective CRF(1) antagonists. The synthesis, structure-activity relationships and pharmacokinetic properties of compounds within this series is presented. PMID- 15341490 TI - Dipeptidomimetic ketomethylene isosteres as pro-moieties for drug transport via the human intestinal di-/tripeptide transporter hPEPT1: design, synthesis, stability, and biological investigations. AB - Five dipeptidomimetic-based model prodrugs containing ketomethylene amide bond replacements were synthesized from readily available alpha,beta-unsaturated gamma ketoesters. The model drug (BnOH) was attached to the C-terminus or to one of the side chain positions of the dipeptidomimetic. The stability, the affinity for the di-/tripeptide transporter hPEPT1, and the transepithelial transport properties of the model prodrugs were investigated. ValPsi[COCH(2)]Asp(OBn) was the compound with highest chemical stability in buffers at pH 6.0 and 7.4, with half-lives of 190 and 43 h, respectively. All five compounds showed high affinity for hPEPT1 (K(i) values < 1 mM), and PhePsi[COCH(2)]Asp(OBn) and ValPsi[COCH(2)]Asp(OBn) had the highest affinities with K(i) values of 68 and 19 microM, respectively. An hPEPT1-mediated transport component was demonstrated for the transepithelial transport of three compounds, a finding that was corroborated by hPEPT1-mediated intracellular uptake. The results indicate that the stabilized Phe-Asp and Val Asp derivatives are promising pro-moieties in a prodrug approach targeting hPEPT1. PMID- 15341491 TI - 2-Pyrazolyl-N(6)-substituted adenosine derivatives as high affinity and selective adenosine A(3) receptor agonists. AB - We describe the synthesis of new high affinity and selective A(3)-adenosine receptor (A(3)-AdoR) agonists. Introduction of a methyl group at the N(6) position of the A(2A)-AdoR selective 2-pyrazolyl-adenosine analogues (Figure 2) brought about a substantial increase in the A(3)-AdoR binding affinity and selectivity. While the N(6)-desmethyl analogues 3a and 4 were inactive at the A(3)-AdoR (K(i) > 10 microM), the corresponding N(6)-methyl analogues 5 and 22 showed good binding affinity at the A(3)-AdoR (K(i) = 73 and 97 nM, respectively). Replacement of the carboxamide group in 5 with different heteroaryl groups resulted in analogues with high affinities and selectivity for the A(3)-AdoR. (2R,3S,4R)-tetrahydro-2-(hydroxymethyl)-5-(6-(methylamino)-2-(4 (pyridin-2-yl)-1H-pyrazol-1-yl)-9H-purin-9-yl)furan-3,4-diol (15, K(i) = 2 nM) displayed high selectivity for the A(3)-AdoR versus A(1)- and A(2A)-AdoRs (selectivity ratios of 1900 and >2000, respectively). PMID- 15341492 TI - Synthesis and biological activity of analogues of the antimicrotubule agent N,beta,beta-trimethyl-L-phenylalanyl-N(1)-[(1S,2E)-3-carboxy-1-isopropylbut-2 enyl]- N(1),3-dimethyl-L-valinamide (HTI-286). AB - Hemiasterlin, a tripeptide isolated from marine sponges, induces microtubule depolymerization and mitotic arrest in cells. HTI-286, an analogue from an initial study of the hemiasterlins, is presently in clinical trials. In addition to its potent antitumor effects, 2 has the advantage of circumventing the P glycoprotein-mediated resistance that hampers the efficacy of other antimicrotubule agents such as paclitaxel and vincristine in animal models. This paper describes an in-depth study of the structure--activity relationships of analogues of 2, their effects on microtubule polymerization, and their in vitro and in vivo anticancer activity. Regions of the molecule necessary for potent activity are identified. Groups tolerant of modification, leading to novel analogues, are reported. Potent analogues identified through in vivo studies in tumor xenograft models include one superior analogue, HTI-042. PMID- 15341493 TI - Design of 2,5-dimethyl-3-(6-dimethyl-4-methylpyridin-3-yl)-7 dipropylaminopyrazolo[1,5-a]pyrimidine (NBI 30775/R121919) and structure- activity relationships of a series of potent and orally active corticotropin releasing factor receptor antagonists. AB - We have previously shown that 3-phenylpyrazolo[1,5-a]pyrimidines exemplified by 8 were potent antagonists of the human corticotropin-releasing factor-1 receptor. A series of 3-pyridylpyrazolo[1,5-a]pyrimidines 15, 25-30, 34, and 35 containing a weakly basic pyridine ring at the 3-position of the bicyclic nucleus was designed to reduce lipophilicity from the initial leads such as 7. Here, we showed that these 3-pyridyl compounds exhibited potent antagonists at the human CRF(1) receptor. Moreover, the hydrophilic and weakly basic pyridine moiety increased the water solubility of some analogues. Compound 26 h exhibited good binding affinity at the human CRF(1) receptor with a K(i) value of 3.5 nM. As a functional antagonist, it dose-dependently inhibited CRF-stimulated cAMP production in cells expressing the CRF(1) receptor (IC(50) = 50 nM), and CRF stimulated ACTH release from cultured rat pituitary cells (IC(50) = 20 nM). 26 h had a log P value of 4.9 and water solubility of greater than 10 mg/mL. Pharmacokinetic studies in rats showed that 26 h was orally bioavailable and able to penetrate into the brain. 26 h has been demonstrated in vivo efficacy in animal behavioral models that measure anxiolytic activity. These results suggest that analogues from this series were potent CRF(1) receptor antagonists with proper physicochemical properties and good pharmacokinetic profiles. 26 h was developed into a clinical compound and exhibited efficacy in patients with major depression. PMID- 15341494 TI - Newer systemic antifungal agents : pharmacokinetics, safety and efficacy. AB - The past few years have seen the advent of several new antifungal agents, including those of a new class and a new generation of an existing class. Caspofungin, the first available echinocandin, has greatly expanded the antifungal armamentarium by providing a cell wall-active agent with candidacidal activity as well as demonstrated clinical efficacy in the therapy of aspergillosis refractory to available therapy. In addition, in clinical trials, caspofungin had comparable efficacy to amphotericin B for candidaemia and invasive Candida infections. Caspofungin and two more recently introduced echinocandins, micafungin and anidulafungin, are available as intravenous formulations only and characterised by potent anti-candidal activity, as well as few adverse events and drug interactions. Voriconazole, the first available second-generation triazole, available in both intravenous and oral formulations, has added a new and improved therapeutic option for primary therapy of invasive aspergillosis and salvage therapy for yeasts and other moulds. In a randomised trial, voriconazole demonstrated superior efficacy and a survival benefit compared with amphotericin B followed by other licensed antifungal therapy. This and data from a noncomparative study led to voriconazole becoming a new standard of therapy for invasive aspergillosis. Voriconazole has several important safety issues, including visual adverse events, hepatic enzyme elevation and skin reactions, as well as a number of drug interactions. Posaconazole, only available orally and requiring dose administration four times daily, shows encouraging efficacy in difficult to treat infections due to zygomycetes. Ravuconazole, available in both intravenous and oral formulations, has broad-spectrum in vitro potency and in vivo efficacy against a wide range of fungal pathogens. Clinical studies are underway. Despite the advances offered with each of these drugs, the morbidity and mortality associated with invasive fungal infections remains unacceptable, especially for the most at-risk patients. For individuals with severe immunosuppression as a result of chemotherapy, graft-versus-host disease and its therapy, or transplantation, new drugs and strategies are greatly needed. PMID- 15341495 TI - Early-stage multiple sclerosis : what are the treatment options? AB - Converging evidence from recent years indicates that the current attitude to the use of immunomodulatory drugs in the treatment of multiple sclerosis (MS) may be too conservative. This evidence originates from studies of the pathophysiology and pathology of MS, magnetic resonance imaging (MRI) studies and clinical trials. Several studies have shown that antigen spreading and propagation of self recognition seem to occur during the clinical progression of MS. The immunopathology may change during the course of disease. Primary selective demyelination can be followed by a secondary oligodendrocyte loss and remyelination becomes less effective. MRI studies have shown that patients with clinically isolated syndromes, who at presentation have more than a few brain lesions on MRI, have a high risk of disease progression over a period of 5-10 years. The most direct evidence comes from two placebo-controlled trials of interferon-beta in very early MS. A delay in time to conversion to clinically definite MS and a significant decrease in MRI activity support an early stage treatment strategy. Taken together, the evidence indicates that treatment with immunomodulatory therapy should be started at an early stage in patients with a high-risk profile for further disease activity, although this may result in over treatment of a small number of patients. However, further prolonged studies are needed to investigate the long-term benefit of early-stage treatment in MS. PMID- 15341496 TI - Antiviral therapy for influenza : a clinical and economic comparative review. AB - Each year influenza epidemics cause a considerable burden of disease. Vaccination against influenza A and B viruses has been and remains the cornerstone of influenza prevention, but antiviral therapy can serve as an important adjunct to vaccination in controlling the impact of the disease. Two classes of drugs are currently licensed in a large number of countries for the treatment of influenza. The M2 ion channel blockers or amantadanes (amantadine and rimantadine) are specific inhibitors of influenza A virus replication, whereas the neuraminidase inhibitors (zanamivir and oseltamivir) are active against influenza A and B viruses. Readily transmissible drug-resistant viruses develop frequently during amantadane treatment but not during neuraminidase inhibitor treatment. In this review, efficacy and safety data from randomised controlled trials are evaluated to gain an understanding of what we can and cannot expect from antiviral treatment. All four drugs shorten the course of influenza disease by approximately 1 day and relieve symptoms to some extent, but there is still uncertainty as to whether antiviral therapy leads to a reduction of serious complications and hospitalisation. The results of cost-effectiveness analyses are very diverse, in part because of differences in methodology but also because there is no consensus on what probabilities to assign to the key risks and benefits that form the basis of these studies. Consensus statements by advisory bodies in England and Germany recommend neuraminidase inhibitors for the therapy of influenza in high-risk individuals such as people over 65 years or under 2 years, and individuals with chronic cardiovascular, pulmonary or renal disease, diabetes mellitus or immunosuppression. However, there is no agreement as to whether antiviral therapy can be generally recommended for otherwise healthy children and adults. The availability of safe and effective antiviral therapy options should be kept in mind by the practising clinician, while more specific recommendations and policy formulation will depend on additional efficacy data that include frequency of complications and hospitalisation as outcome measures. PMID- 15341498 TI - Emtricitabine/tenofovir disoproxil fumarate. AB - The nucleoside analogue reverse transcriptase inhibitor (RTI) emtricitabine and the nucleotide analogue RTI tenofovir disoproxil fumarate (tenofovir DF) have each shown antiviral activity against a number of HIV clinical isolates and cell lines. HIV variants with reduced susceptibility to emtricitabine and tenofovir have been selected for in vitro and have also been isolated from patients receiving the agents. Low rates of these variants have been observed in patients experiencing virological failure in large studies of emtricitabine- or tenofovir DF-containing therapy. Co-formulated oral emtricitabine/tenofovir DF was bioequivalent to the two agents as separate formulations in a pharmacokinetic trial in healthy volunteers. There are no published data on the clinical antiviral efficacy of co-formulated oral emtricitabine/tenofovir DF. However, each agent is effective in combination regimens with other drugs. Ongoing studies in antiretroviral-naive patients are evaluating the efficacy of the individual formulations given together in combination with efavirenz or lopinavir/ritonavir. In the latter trial, HIV RNA levels were reduced and CD4+ cell counts were increased at 24 and 48 weeks. Emtricitabine and tenofovir DF are generally well tolerated. Diarrhoea, nausea and vomiting were the most common adverse events reported with coadministered emtricitabine and tenofovir DF as separate formulations as part of combination therapy. PMID- 15341501 TI - Intravenous lansoprazole: in erosive oesophagitis. AB - An intravenous formulation of lansoprazole, a proton pump inhibitor, is approved for use in patients with erosive oesophagitis who are temporarily unable to take oral lansoprazole. In healthy volunteers, oral and intravenous lansoprazole 30 mg/day were equivalent in suppressing basal and pentagastrin-stimulated maximum gastric acid output. Moreover, the mean 24-hour intragastric pH did not differ significantly following oral or intravenous administration of lansoprazole and was significantly higher with both formulations than with intravenous polyethylene glycol vehicle. After treatment for 7 days in patients with erosive oesophagitis, intravenous lansoprazole (30 mg/day) recipients had significantly lower median stimulated and basal gastric acid output measurements than placebo recipients. Median pentagastrin-stimulated gastric acid output levels were equivalent after 7 days treatment with intravenous or oral lansoprazole. Intravenous lansoprazole is generally well tolerated. All adverse events experienced by patients with erosive oesophagitis who received intravenous lansoprazole were mild or moderate in severity. PMID- 15341497 TI - Effect of immunosuppressive agents on long-term survival of renal transplant recipients: focus on the cardiovascular risk. AB - In the control of acute rejection, attention is being focused more and more on the long-term adverse effects of the immunosuppressive agents used. Since cardiovascular disease is the main cause of death in renal transplant recipients, optimal control of cardiovascular risk factors is essential in the long-term management of these patients. Unfortunately, several commonly used immunosuppressive drugs interfere with the cardiovascular system. In this review, the cardiovascular adverse effects of the immunosuppressive agents currently used for maintenance immunosuppression are thoroughly discussed. Optimising immunosuppression means finding a balance between efficacy and safety. Corticosteroids induce endothelial dysfunction, hypertension, hyperlipidaemia and diabetes mellitus, and impair fibrinolysis. The use of corticosteroids in transplant recipients is undesirable, not only because of their cardiovascular effects, but also because they induce such adverse effects as osteoporosis, obesity, and atrophy of the skin and vessel wall. Calcineurin inhibitors are the most powerful agents for maintenance immunosuppression. The calcineurin inhibitor ciclosporin (cyclosporine) not only induces these same adverse effects as corticosteroids but is also nephrotoxic. Tacrolimus has a more favourable cardiovascular risk profile than ciclosporin and is also less nephrotoxic. It has little or no effect on blood pressure and serum lipids; however, its diabetogenic effect is more prominent in the period immediately following transplantation, although at maintenance dosages, the diabetogenic effect appears to be comparable to that of ciclosporin. The diabetogenic effect of tacrolimus can be managed by reducing the dose of tacrolimus and early corticosteroid withdrawal. The effect of tacrolimus on endothelial function has not been completely elucidated. The proliferation inhibitors azathioprine and mycophenolate mofetil (MMF) have little effect on the cardiovascular system. Yet, indirectly, by inducing anaemia, they may lead to left ventricular hypertrophy. MMF is an attractive alternative to azathioprine because of its higher potency and possibly lower risk of malignancies. Sirolimus also induces anaemia, but may be promising because of its antiproliferative features. Whether the hyperlipidaemia induced by sirolimus counteracts its beneficial effects is, as yet, unknown. It may be combined with MMF, however, initial attempts resulted in severe mouth ulcers. PMID- 15341504 TI - Brivudin (bromovinyl deoxyuridine). AB - Brivudin is an oral thymidine analogue indicated for the early treatment of acute herpes zoster in immunocompetent adults. It has high, selective activity against varicella zoster virus (VZV), inhibiting VZV replication, possibly through competitive inhibition of viral DNA polymerase, or by acting as an alternative substrate to deoxythymidine triphosphate, causing viral DNA strand breakage. In a large, 7-day, phase III trial in immunocompetent patients with herpes zoster, once-daily brivudin 125mg was significantly more effective than oral acyclovir 800mg five times daily in reducing the mean time from start of treatment to last vesicular eruption, and was as effective as acyclovir at healing lesions and alleviating acute zoster-related pain. The likelihood of developing post-herpetic neuralgia (PHN) in immunocompetent patients aged > or =50 years was significantly lower with brivudin than with acyclovir. Brivudin was as effective as oral famciclovir 250mg three times daily in terms of the prevalence of PHN, the time to last vesicular eruption and lesion healing in another large, 7-day, phase III study in immunocompetent patients with herpes zoster. Oral brivudin is generally well tolerated, with a similar tolerability profile to those of oral acyclovir or famciclovir. Nausea was the most commonly reported adverse event. PMID- 15341507 TI - Fosamprenavir: a review of its use in the management of antiretroviral therapy naive patients with HIV infection. AB - Fosamprenavir (GW433908, Lexiva, Telzir) is an oral prodrug of the protease inhibitor (PI) amprenavir, with a reduced daily pill burden. Fosamprenavir, in combination with other antiretroviral agents, is indicated for the treatment of patients with HIV infection, particularly those who have not previously received antiretroviral therapy. Viral load reductions were at least as great with fosamprenavir-based regimens as those achieved with nelfinavir-based regimens in two large, 48-week, randomised, multicentre trials in antiretroviral therapy naive patients with HIV infection. In the NEAT study, more patients receiving twice-daily fosamprenavir in combination with abacavir and lamivudine achieved HIV RNA levels <400 copies/mL than those receiving a similar nelfinavir-based regimen. Results of the SOLO study showed similar reductions in viral load among patients who received once-daily ritonavir-boosted fosamprenavir and those treated with twice-daily nelfinavir, both in combination with twice-daily abacavir and lamivudine. In both trials, virological failure rates were at least twice as high with the nelfinavir-based regimen as they were with the fosamprenavir-based regimen. Fosamprenavir was generally well tolerated in clinical trials. The most common adverse events among patients treated with fosamprenavir, with or without ritonavir, plus abacavir and lamivudine were diarrhoea, nausea, vomiting, abdominal pain, drug hypersensitivity and skin rash. The incidence of diarrhoea was significantly lower with fosamprenavir-based therapy than with nelfinavir-based therapy in the NEAT and SOLO trials. The resistance profile of fosamprenavir is consistent with that of amprenavir. Amprenavir-resistant viral isolates from patients experiencing treatment failure with fosamprenavir-based therapy in the NEAT study showed little or no cross resistance to several other PIs, and protease mutations commonly selected for by various other PIs were not observed. In the SOLO study, protease resistance mutations were not observed in viral isolates from patients experiencing treatment failure with ritonavir-boosted fosamprenavir-based therapy. In conclusion, fosamprenavir-based regimens have shown good antiviral efficacy and are generally well tolerated in antiretroviral therapy-naive patients with HIV infection. Available data on the resistance profile of the drug suggest that it may be used early in the course of therapy without compromising a range of future treatment options. The relatively low pill burden and lack of food restrictions with fosamprenavir may improve adherence to therapy. Further studies are needed to compare fosamprenavir with other PIs and to establish the long-term efficacy of fosamprenavir-based regimens. In conclusion, fosamprenavir appears to be a promising agent for the treatment of antiretroviral therapy-naive patients with HIV infection. PMID- 15341508 TI - Cabergoline : a review of its use in the treatment of Parkinson's disease. AB - Cabergoline is a synthetic ergoline dopamine agonist with a high affinity for dopamine D2 receptors and a long elimination half-life. This agent provides continuous dopaminergic stimulation with once-daily administration. Adjuvant oral cabergoline is usually well tolerated and effective in controlling symptoms in patients with advanced Parkinson's disease experiencing response fluctuations to long-term levodopa therapy. In patients with early Parkinson's disease, cabergoline (with or without levodopa) is well tolerated and effective in controlling disease symptoms, and may reduce the risk of developing drug-induced motor complications. Data from two pharmacoeconomic analyses suggest that cabergoline may be a cost-effective treatment option versus levodopa in patients with early Parkinson's disease, and highlight the need for further evaluation of the drug in this indication. PMID- 15341509 TI - Transgenic expression of human complement regulators reduces skeletal muscle ischaemia/reperfusion injury in mice. AB - This study aimed to explore the hypothesis that activated complement components contribute significantly to I/R (ischaemia/reperfusion) injury in skeletal muscle. After 50, 70 and 90 min of tourniquet ischaemia and 24 h of reperfusion, viability of the medial gastrocnemius muscle in CBA-C57BL/6 wild-type mice, assessed histochemically by reduction of NBT (Nitro Blue Tetrazolium) dye, was 60, 21 and 8% respectively. Skeletal muscle viability after 70 min of ischaemia and 24 h of reperfusion in transgenic mice expressing a combination of human CD46, CD55 and CD59, all inhibitors of complement activation, was 45% compared with 24% in ischaemic reperfused wild-type mice (P=0.008; n=6 per group). Muscle from sham-treated transgenic mice and wild-type littermates had no significant loss of viability relative to normal contralateral gastrocnemius muscle. A significant reduction in myeloperoxidase activity (a measure of neutrophil infiltration), xanthine oxidase activity (a source of free radicals) and water content (a measure of oedema) was observed in ischaemic reperfused muscle from transgenic mice compared with ischaemic reperfused wild-type muscle (P<0.05). Haematoxylin and eosin-stained histological sections also showed less damage and less apparent leucocyte infiltration in muscles from ischaemic reperfused transgenic mice than those from wild-type animals given the same degree of injury. Muscles from sham-treated transgenic and wild-type controls were almost identical with normal muscle. It is concluded that complement activation contributes to the pathogenesis of I/R injury in murine skeletal muscle, resulting in increased neutrophil infiltration into the injured muscle, increased free radical production and vascular permeability during reperfusion, and a net detrimental effect on muscle viability. PMID- 15341510 TI - Effect of ischaemic preconditioning on hepatic oxygenation, microcirculation and function in a rat model of moderate hepatic steatosis. AB - IPC (ischaemic preconditioning) may protect the steatotic liver, which is particularly susceptible to I/R (ischaemia/reperfusion) injury. Hepatic steatosis was induced in Sprague-Dawley rats with a high-cholesterol (2%) diet for 12 weeks after which rats were subjected to I/R (ischaemia/reperfusion; 45 min of lobar ischaemia followed by 2 h of reperfusion). Rats were divided into three study groups (n=6 each) receiving: (i) sham laparotomy alone, (ii) I/R, and (iii) IPC (5 min of ischaemia, followed by 10 min of reperfusion) before I/R. Hepatic extra and intra-cellular oxygenation and HM (hepatic microcirculation) were measured with near-infrared spectroscopy and laser Doppler flowmetry respectively. Plasma liver enzymes and hepatic tissue ATP were measured as markers of liver injury. Histology showed moderate-grade steatosis in the livers. At the end of 2 h of reperfusion, I/R significantly decreased extra- and intra-cellular oxygenation concomitant with a failure of recovery of HM (21.1+/-14.4% of baseline; P<0.001 compared with sham animals). IPC increased intracellular oxygenation (redox state of the copper centre of cytochrome oxidase; P<0.05 compared with rats receiving I/R alone) and flow in HM (70.9+/-17.1% of baseline; P<0.001 compared with rats receiving I/R alone). Hepatocellular injury was significantly reduced with IPC compared with I/R injury alone (alanine aminotransferase, 474.8+/-122.3 compared with 5436.3+/-984.7 units/l respectively; P<0.01; aspartate aminotransferase, 630.8+/-76.9 compared with 3166.3+/-379.6 units/l respectively; P<0.01]. In conclusion, IPC has a hepatoprotective effect against I/R injury in livers with moderate steatosis. These data may have important clinical implications in liver surgery and transplantation. PMID- 15341511 TI - Diabetic cardiomyopathy: mechanisms, diagnosis and treatment. AB - Independent of the severity of coronary artery disease, diabetic patients have an increased risk of developing heart failure. This clinical entity has been considered to be a distinct disease process referred to as 'diabetic cardiomyopathy'. Experimental studies suggest that extensive metabolic perturbations may underlie both functional and structural alterations of the diabetic myocardium. Translational studies are, however, limited and only partly explain why diabetic patients are at increased risk of cardiomyopathy and heart failure. Although a range of diagnostic methods may help to characterize alterations in cardiac function in general, none are specific for the alterations in diabetes. Treatment paradigms are very much limited to interpretation and translation from the results of interventions in non-diabetic patients with heart failure. This suggests that there is an urgent need to conduct pathogenetic, diagnostic and therapeutic studies specifically in diabetic patients with cardiomyopathy to better understand the factors which initiate and progress diabetic cardiomyopathy and to develop more effective treatments. PMID- 15341512 TI - Mitochondrial mechanisms of neural cell apoptosis. AB - The importance of calcium overload, mitochondrial dysfunction, and free radical generation to neuropathological processes has been recognized for many years. Only more recently has evidence accumulated that the programmed cell death process of apoptosis plays an integral role not only in the development of the nervous system, but in the loss of cells following acute neurological insults and chronic disease. In 1996 came the landmark discovery that cytochrome c, an evolutionary old and essential component of the respiratory chain, has a second and deadly function when it escapes the mitochondrion: triggering the cell death cascade. A flurry of activity has since ensued in an effort to understand the mechanistic events associated with mitochondrial permeabilization during apoptosis and regulation by an enigmatic family of proteins characterized by homology to the proto-oncogene Bcl-2. This review discusses the evidence for various release mechanisms of apoptotic proteins (e.g. cytochrome c) from neural cell mitochondria, focusing particularly on roles for calcium, Bax, p53, and oxidative stress. The need for new drugs that act at the level of the mitochondrion to prevent apoptosis is also highlighted. PMID- 15341513 TI - Functional dissociation of apelin receptor signaling and endocytosis: implications for the effects of apelin on arterial blood pressure. AB - Apelin is a novel neuropeptide involved in the regulation of body fluid homeostasis and cardiovascular functions. It acts through a G protein-coupled receptor, the APJ receptor. We studied the structure-activity relationships of apelin at the rat apelin receptor, tagged at its C-terminal end with enhanced green fluorescent protein and stably expressed in CHO cells. We evaluated the potency of N- and C-terminal deleted fragments of K17F to bind with high affinity to the apelin receptor, and to inhibit cAMP production and to induce apelin receptor internalization. We first characterized the internalization and trafficking of the rat apelin receptor. This receptor was internalized via a clathrin-dependent mechanism and our results suggest that receptor trafficking may follow a recycling pathway. We then tried to identify the amino acids of K17F required for apelin activity. The first five N-terminal and the last two C terminal amino acids of K17F were not essential for apelin binding or the inhibition of cAMP production. However, the full-length sequence of K17F was the most potent inducer of apelin receptor internalization because successive N terminal amino-acid deletions progressively reduced internalization and the removal of a single amino acid at the C-terminus abolished this process. Finally, the most novel observation of this work is that hypotensive actions of apelin peptides correlate best with the ability of those ligands to internalize. Thus, apelin receptor signaling and endocytosis are functionally dissociated, possibly reflecting the existence of several conformational states of this receptor, stabilized by the binding of different apelin fragments to the apelin receptor. PMID- 15341514 TI - Molecular aging of tau: disulfide-independent aggregation and non-enzymatic degradation in vitro and in vivo. AB - Smearing from high-molecular-mass regions to low-molecular-mass regions on western blot is the most striking observation of the tau making up paired helical filaments in brain tissues affected by Alzheimer's disease. Because our previous study showed site-specific deamidation/isomerization in the smeared tau in vivo, a feature of protein aging, recombinant tau was subjected to prolonged (up to 90 days) in vitro incubation. Carboxymethylated tau at approximately 50 kDa gradually disappeared and was converted to dimers and to high- and low-molecular mass smearing. In addition, the same site-specific deamidation/isomerization as previously identified in the smeared tau in vivo emerged. Most importantly, tau was spontaneously degraded, generating fragments that start from bulky residues next to asparaginyl residues. This spontaneous degradation of tau probably represents non-enzymatic cleavage through the formation of succinimide intermediates. Similar degradation products starting from the bulky residues next to asparaginyl residues were found in the smeared tau in vivo partially purified from the homogenates from Alzheimer's disease brains. PMID- 15341515 TI - Conserved residues within the putative active site of gamma-secretase differentially influence enzyme activity and inhibitor binding. AB - Gamma-secretase performs the final processing step in the generation of amyloid beta (Abeta) peptides, which are believed to be causative for Alzheimer's disease. Presenilins (PS) are required for gamma-secretase activity and the presence of two essential intramembranous aspartates (D257 and D385) has implicated this region as the putative catalytic centre of an aspartyl protease. The presence of several key hydrogen-bonding residues around the active site of classical aspartyl proteases led us to investigate the role of both the critical aspartates and two nearby conserved hydrogen bond donors in PS1. Generation of cell lines stably overexpressing the D257E, D385E, Y256F and Y389F engineered mutations has enabled us to determine their role in enzyme catalysis and binding of a transition state analogue gamma-secretase inhibitor. Here we report that replacement of either tyrosine residue alters gamma-secretase cleavage specificity, resulting in an increase in the production of the more pathogenic Abeta42 peptide in both cells and membranous enzyme preparations, without affecting inhibitor binding. In contrast, replacement of either of the aspartate residues precludes inhibitor binding in addition to inactivation of the enzyme. Together, these data further incriminate the region around the intramembranous aspartates as the active site of the enzyme, targeted by transition state analogue inhibitors, and highlight the roles of individual residues. PMID- 15341516 TI - Cerebral creatine kinase deficiency influences metabolite levels and morphology in the mouse brain: a quantitative in vivo 1H and 31P magnetic resonance study. AB - Creatine kinase (CK)-catalysed ATP-phosphocreatine (PCr) exchange is considered to play a key role in energy homeostasis of the brain. This study assessed the metabolic and anatomical consequences of partial or complete depletion of this system in transgenic mice without cytosolic B-CK (B-CK-/-), mitochondrial ubiquitous CK (UbCKmit-/-), or both isoenzymes (CK -/-), using non-invasive quantitative magnetic resonance (MR) imaging and spectroscopy. MR imaging revealed an increase in ventricle size in a subset of B-CK-/- mice, but not in animals with UbCKmit or compound CK mutations. Mice lacking single CK isoenzymes had normal levels of high-energy metabolites and tissue pH. In the brains of CK double knockouts pH and ATP and Pi levels were also normal, even though PCr had become completely undetectable. Moreover, a 20-30% decrease was observed in the level of total creatine and a similar increase in the level of neuronal N-acetyl aspartate compounds. Although CKs themselves are not evenly distributed throughout the CNS, these alterations were uniform and concordant across different brain regions. Changes in myo-inositol and glutamate peaks did appear to be mutation type and brain area specific. Our results challenge current models for the biological significance of the PCr-CK energy system and suggest a multifaceted role for creatine in the brain. PMID- 15341517 TI - Cloning and expression of the mouse histamine H3 receptor: evidence for multiple isoforms. AB - The existence of mouse H3-receptor isoforms was investigated by PCR analysis and cDNA cloning. Splicing mechanisms previously reported in various species are conserved in the mouse. The retention/deletion of a fragment in the third intracellular loop of the mouse receptor leads to the existence of three isoforms designated mH(3(445)), mH(3(413)) and mH(3(397)) according to the length of their deduced amino acid sequence. PCR analysis showed that mouse H3-receptor isoforms display different expression patterns in the brain. Following expression in Cos-1 cells, [125I]iodoproxyfan binding indicated similar pharmacological profiles of the mH(3(445)), mH(3(413)) and mH(3(397)) isoforms. The pharmacological profile of the mouse H3 receptor is more similar to the rat receptor than to the human receptor, although some differences were also observed between the mouse and rat receptors. For example, the potency of thioperamide and ciproxifan is slightly higher at the mouse receptor than at the rat receptor but 40-100-fold higher than at the human receptor. In situ hybridization histochemistry showed that the distribution of H3-receptor mRNAs in the mouse brain is rather similar to that previously reported in the rat brain. However, the autoradiographic and cellular expression patterns observed in several brain areas such as the thalamus or hippocampus reveal important differences between the two species. PMID- 15341518 TI - Erk1/2 but not PI3K pathway is required for neurotrophin 3-induced oligodendrocyte differentiation of post-natal neural stem cells. AB - Neurotrophin 3 (NT3) induces mouse cortical stem cells to an asymmetric division from a symmetric division, suggesting that NT3 may work as an early differentiative signal for neural stem cells (NSCs). Here, using cultured post natal hippocampal stem cells as a model, we demonstrated that NT3-stimulation causes NSCs to differentiate into oligodendrocyte precursors (OLPs) through an extracellular signal-related kinase1/2 (Erk1/2)-dependent pathway. Following the treatment of NT3 for 24 h, NSCs differentiated into more OLPs and fewer neurons, whereas the proliferation and survival of OLPs were not affected. NT3 induced a series of intracellular responses including enhancement of phosphorylation of Erk1/2 or Akt and increase of expression of oligodendrocyte lineage gene (Olig) 1, a transcriptional factor known to participate in oligodendrocyte development. Application of U0126, a specific inhibitor of MEK1/2 which are upstream to Erk1/2, blocked the phosphorylation of Erk1/2, suppressed the expression of Olig 1 and prevented NSC differentiation into OLPs in response to NT3 stimulation. Blockade of TrkC also inhibited the differentiation of NSCs to OLPs induced by NT3. However, administration of LY294002, an inhibitor of phosphatidylinositol 3 kinase (PI3K), blocked the phosphorylation of Akt but did not affect the effect of NT3 on the expression of Olig-1 and on NSC differentiation into OLPs. Taken together, these results suggest that NT3 induce NSCs to differentiate into OLPs by enhancing the expression of Olig-1 through an Erk1/2-dependent pathway. PMID- 15341519 TI - Differential gene expression induced by chronic levodopa treatment in the striatum of rats with lesions of the nigrostriatal system. AB - Levodopa, the major treatment for patients with Parkinson's disease, has been shown to induce a variety of compensatory effects, including facilitation of sprouting by dopaminergic neurons, in experimental animals with lesions leading to denervation of the striatum. To better understand the cellular and molecular environment where most of these compensatory changes take place, in particular elements that might contribute to the recovery of dopaminergic innervation, we have constructed a differential expression library enriched in transcripts from the striata of rats with lesions of the medial forebrain bundle treated with levodopa for 6 months. We have used this library to screen an expression array of rat genes representing the major cell functions, and have identified several that are involved in neurotrophic mechanisms and plasticity. We have confirmed the differential expression of selected transcripts by non-radioactive in situ hybridization, and report that the growth factor pleiotrophin, myelin basic protein and calmodulin are overexpressed in the denervated striatum of levodopa treated rats. PMID- 15341520 TI - Apoptotic cell death influences the signaling activity of the amyloid precursor protein through ShcA and Grb2 adaptor proteins in neuroblastoma SH-SY5Y cells. AB - The amyloid precursor protein (APP) is an ubiquitous receptor-like molecule involved in the pathogenesis of Alzheimer's disease (AD). APP and some of its C terminal proteolytic fragments (CTFs) have been shown to be phosphorylated and to interact with cytosolic phosphotyrosine binding (PTB) domain containing proteins involved in cell signaling and vesicular transport. Among others, the interaction between tyrosine-phosphorylated CTFs and ShcA-Grb2 adaptors is highly enhanced in AD brain. Here we have identified in SH-SY5Y neuroblastoma cells an interaction between APP holoprotein and the adaptor Grb2. Upon activation of apoptotic cell death this interaction is rapidly degraded, APP is partially cleaved and the complex APP/Grb2 is replaced by a new complex between CTFs and ShcA that still involves Grb2. The formation of these complexes is regulated by beta-site APP cleaving enzyme 1 and influences the phosphorylation of mitogen-activated protein kinase p44/42 extracellular signal-regulated kinase as well as the level of apoptotic death of the cells. These data suggest a dual role in cell signaling for APP and its CTFs in neuroblastoma cells, in a manner similar to that previously reported for other tyrosine kinase receptor, through a tightly regulated coupling with alternative intracellular adaptors to control the signaling of the cell. PMID- 15341521 TI - Modulation of extracellular signal-regulated kinase (ERK) activity by acute and chronic opioid treatment in neuronal and glial cell lines. AB - Acute mu opioid application has been shown to activate extracellular signal related kinases (ERKs) in various non-neural cell lines. However, ERK activation in neuronal cells following acute morphine treatment is more questionable. Moreover, the ERK activation phenomenon observed in vivo after withdrawal of chronic opioids has never been demonstrated in vitro. The goal of this study was to determine if mu agonist treatment induced ERK activation acutely or after withdrawal of chronic opioids in one glial and three neuronal cell lines. We found that acute application of opioids was not able to activate ERK in neuronal cell lines but was able to activate ERK in a glial cell line. In another set of experiments, cells were chronically treated with escalating doses of a mu opioid agonist. After 8 days, the agonist was removed from the media and naloxone applied. Acute ERK activation was not seen in any tested cell line after agonist removal. These findings suggest that opioids may acutely activate ERK in non neuronal cells, and that the acute ERK activation observed in some brain regions during opioid withdrawal in vivo might be mediated by indirect effects on neuronal cells. PMID- 15341522 TI - Effects of psychotomimetic and antipsychotic agents on neocortical and striatal concentrations of various amino acids in the rat. AB - A subcutaneous injection of small and moderate doses (1.6, 3.2, 4.0 and 4.8 mg/kg) of the schizophrenomimetic methamphetamine caused a dose-related increase in the tissue content (the net content) of L-Arg and L-Asn in the neocortex and striatum at 60 min, but not at 360 min, after injection. The methamphetamine induced (4.8 mg/kg) increases in levels of these amino acids were significantly attenuated by pretreatment with an antipsychotic drug, haloperidol (1 mg/kg) or clozapine (10 mg/kg). In the neocortex, a clozapine-reversible increase in the level of L-Thr was also observed 60 min after methamphetamine administration. Striatal concentrations of L-Glu, L-Ser, LThr, Gly and L-Ala were augmented by the same regimen in a haloperidol- and clozapine-sensitive fashion. A moderate dose of another schizophrenomimetic phencyclidine (7.5 mg/kg) given subcutaneously induced robust abnormal behavior, a diminution in the neocortical and striatal levels of L-Asp and an increase in the striatal L-Ala content without significant effects on the other amino acids studied. These results suggest that neocortical and striatal L-Arg, L-Asn, L-Thr, Gly, L-Ala or L-Ser may be implicated in the psychotomimetic effects of methamphetamine and might display mutual interaction with cerebral dopaminergic transmission. The differential effects of methamphetamine and phencyclidine on the net neocortical and striatal concentrations of various amino acids might, at least in part, underlie the distinct features of psychoses induced by these two drugs. PMID- 15341523 TI - Developmental expression of receptor for advanced glycation end products (RAGE), amphoterin and sulfoglucuronyl (HNK-1) carbohydrate in mouse cerebellum and their role in neurite outgrowth and cell migration. AB - Receptor for advanced glycation end products (RAGE) has been proposed as a signal transduction receptor to promote neurite outgrowth and cell migration, by its interaction with a neurite outgrowth promoting protein, Amphoterin. Amphoterin has been shown to interact with sulfoglucuronyl carbohydrate (SGC). The developmental expression of RAGE, Amphoterin and SGC was studied in pre-natal and post-natal mouse cerebellum to establish their cellular and subcellular localization and function. The amount of RAGE in the cerebellum increased with age. RAGE was expressed pre-natally in the external germinal layer and post natally in the plasma membranes of the granule neurons of the external and internal granule cell layers and in Purkinje cells. Immunocytochemical analysis by high magnification confocal microscopy showed that RAGE was co-expressed with Amphoterin and SGC in the cell surfaces of granule neurons. This co-localization of RAGE, Amphoterin, and SGC was confirmed in isolated and cultured granule neurons and in migrating granule neurons in explant cultures. Anti-RAGE antibodies inhibited neurite outgrowth and cell migration in explant and slice cultures, similar to anti-Amphoterin and anti-SGC antibodies shown previously. The results suggest that RAGE could act as a signaling molecule for neurite outgrowth and cell migration by its interaction with Amphoterin and that of Amphoterin with SGC. PMID- 15341524 TI - PEN-2 enhances gamma-cleavage after presenilin heterodimer formation. AB - The presenilin (PS) complex, including PS, nicastrin, APH-1 and PEN-2, is essential for gamma-secretase activity, which is required for amyloid beta protein (Abeta) generation. However, the precise individual roles of the three cofactors in the PS complex in Abeta generation remain to be clarified. Here, to distinguish the roles of PS cofactors in gamma-secretase activity from those in PS endoproteolysis, we investigated their roles in the gamma-secretase activity reconstituted by the coexpression of PS N- and C-terminal fragments (NTF and CTF) in PS-null cells. We demonstrate that the coexpression of PS1 NTF and CTF forms the heterodimer and restores Abeta generation in PS-null cells. The generation of Abeta was saturable at a certain expression level of PS1 NTF/CTF, while the overexpression of PEN-2 alone resulted in a further increase in Abeta generation. Although PEN-2 did not enhance PS1 NTF/CTF heterodimer formation, PEN-2 expression reduced the IC50 of a specific gamma-secretase inhibitor, a transition state analogue, for Abeta generation, suggesting that PEN-2 expression enhances the affinity or the accessibility of the substrate to the catalytic site. Thus, our results strongly suggest that PEN-2 is not only an essential component of the gamma-secretase complex but also an enhancer of gamma-cleavage after PS heterodimer formation. PMID- 15341525 TI - Intermediate-conductance calcium-activated potassium channels in enteric neurones of the mouse: pharmacological, molecular and immunochemical evidence for their role in mediating the slow afterhyperpolarization. AB - Calcium-activated potassium channels are critically important in modulating neuronal cell excitability. One member of the family, the intermediate conductance potassium (IK) channel, is not thought to play a role in neurones because of its predominant expression in non-excitable cells such as erythrocytes and lymphocytes, in smooth muscle tissues, and its lack of apparent expression in brain. In the present study, we demonstrate that IK channels are localized on specific neurones in the mouse enteric nervous system where they mediate the slow afterhyperpolarization following an action potential. IK channels were localized by immunohistochemistry on intrinsic primary afferent neurones, identified by their characteristic Dogiel type II morphology. The slow afterhyperpolarization recorded from these cells was abolished by the IK channel blocker clotrimazole. RT-PCR and western analysis of extracts from the colon revealed an IK channel transcript and protein identical to the IK channel expressed in other cell types. These results indicate that IK channels are expressed in neurones where they play an important role in modulating firing properties. PMID- 15341526 TI - Counteraction of axonal growth inhibitory properties of Semaphorin 3A and myelin associated proteins by a synthetic neurotrophic compound. AB - One of the reasons for the lack of nerve regeneration in the CNS is the formation of a glial scar over-expressing multiple inhibitory factors including myelin associated proteins and members of the Semaphorin family. Innovative therapeutic strategies must stimulate axon extension across the lesion site despite this inhibitory molecular barrier. We recently developed a synthetic neurotrophic compound combining an omega-alkanol with a retinol-like cycle (3-(15-hydroxy pentadecyl)-2,4,4,-trimethyl-cyclohexen-2-one (tCFA15)). Here, we demonstrate that tCFA15 is able to promote cortical axon outgrowth in vitro even in the presence of the inhibitory Semaphorin 3A and myelin extracts. This growth promoting effect is selectively observed in axons and requires multiple growth associated intracellular pathways. Our results illustrate the potential use of synthetic neurotrophic compounds to promote nerve regeneration by counteracting the axonal growth inhibition triggered by glial scar-associated inhibitory factors. PMID- 15341527 TI - Mechanism of promoter activity of the beta-amyloid precursor protein gene in different cell lines: identification of a specific 30 bp fragment in the proximal promoter region. AB - The amyloid beta-protein (Abeta) deposited in brains of Alzheimer's disease (AD) patients is proteolytically derived from a large Abeta precursor protein (APP). APP gene expression patterns in the AD brain region indicate that abnormalities of gene regulation may be important in AD pathology. To understand the contribution of different cell types to APP gene expression, we studied it at four levels: promoter activity (by reporter gene assay of transfected cells), DNA nuclear protein interaction (by electrophoretic mobility shift assay), RNA message and protein (by northern and western blotting, respectively). APP mRNA and protein expression levels were greater in neuroblastoma and PC12 cells than in glial or cervix epithelial cells. Relative activity among 12 different promoter regions and within single regions varied according to cell type/cell line. An upstream regulatory region containing a GATA-1 site is necessary for activity in PC12 and glial cells but not in neuroblastoma cells. DNA-protein interactions were examined in three distal and one proximal promoter elements in nuclear extracts belonging to neuronal and non-neuronal cells. The proximal promoter region is important for cell line-specific APP gene expression. Characterization of the APP regulatory region's interaction with cell type specific nuclear factor(s) is important to understand tissue-specific expression of APP seen in AD subjects. PMID- 15341528 TI - In vivo somatic delivery of plasmid DNA and retrograde transport to obtain cell specific gene expression in the central nervous system. AB - We utilised the retrograde transport machinery of neurones to deliver naked plasmid DNA into the central nervous system. A 5.4-kb fragment of the glycine receptor (GlyR) alpha1 subunit gene was cloned and used to drive the expression of a construct encoding for the enhanced green fluorescent protein (EGFP). Injections of the plasmid DNA in the tongue of mice resulted in the expression of the marker protein in hypoglossal motor neurones, showing that the GlyRalpha1 promoter sequence is sufficient to drive expression of the transgene. In order to determine the specificity of expression of the 5.4-kb fragment of the GlyR alpha1 subunit gene promoter, we subsequently injected the plasmid DNA into the mouse central nucleus of the amygdala. This nucleus receives projections from the parabrachial nucleus, a brainstem area that has a high density of GlyRs, and from the insular cortex, a forebrain structure devoid of GlyRs. We observed EGFP labelled neurones in the parabrachial nucleus, but not in the insular cortex, indicating that the 5.4-kb GlyR alpha1 subunit gene promoter confers specificity of expression. This approach provides a simple and rapid way to identify, in vivo, promoter elements that mediate neurone-specific gene expression. PMID- 15341529 TI - Structural determinants for membrane trafficking and G protein selectivity of a mouse olfactory receptor. AB - The G protein-coupled olfactory receptor (OR) superfamily plays a critical role in recognizing a broad range of odorants. Each OR appears to recognize odorants based on similarities in molecular structures such that mOR-EG, a mouse OR, binds eugenol, vanillin, and some other structurally related odorants. Only a few ORs, however, have been characterized functionally due to the difficulties in expressing ORs in heterologous cells. In this report, we demonstrate roles of the N- and C-terminal domains as key elements in the functional expression and signal transducing activity of an OR. Disruption of the N-terminal glycosylation site of the mOR-EG completely impaired its membrane trafficking to the cell surface. Functional expression of the mOR-EG was greatly enhanced by addition of extra N terminal glycosylation sequences. Addition of a C-terminal epitope-tag or C terminal truncation significantly reduced the odorant-response activity, although the receptors were properly targeted to the plasma membrane. Analysis of a series of truncated ORs revealed a region in the C-terminus that was crucial for the receptor activity. Replacement of the C-terminal portion of the mOR-EG with that of rhodopsin disrupted the coupling to G(alphas) but not to G(alpha15), demonstrating that the C-terminus is involved in regulating G protein specificity. These results suggest that glycosylation of the N-terminal portion is critical for OR expression and membrane trafficking, while the C-terminal portion plays a role in defining proper conformation, which, in turn, specifies the G protein selectivity of the OR. This information helps clarify the mechanisms that regulate membrane trafficking and G protein interaction of the OR superfamily. PMID- 15341530 TI - 5-HT stimulates eEF2 dephosphorylation in a rapamycin-sensitive manner in Aplysia neurites. AB - In Aplysia, serotonin mediates behavioral sensitization by increasing the strength of the synapse between sensory and motor neurons, a process known as facilitation. The retention of long-term facilitation is blocked by rapamycin, an inhibitor of a specific translational pathway. One possible rapamycin-sensitive target is the increased translation of 5'-terminal oligopyrimidine mRNAs. These transcripts encode components of the translational machinery and have been proposed to be important for retention of long-term facilitation. We have cloned the 5'-terminal oligopyrimidine mRNA encoding eukaryotic elongation factor 2 and shown that serotonin increased its translation in synaptosomes. Another possible rapamycin-sensitive target is the inactivation of eukaryotic elongation factor 2 kinase. Eukaryotic elongation factor 2 kinase phosphorylates and inactivates eukaryotic elongation factor 2, blocking translational elongation. Serotonin application decreased eukaryotic elongation factor 2 phosphorylation in synaptosomes and in isolated neurites, and this was blocked by rapamycin. We propose a role for the rapamycin-sensitive pathway in neurons. Stimulation blocks translation by inducing calcium entry and phosphorylation of eukaryotic elongation factor 2. This block is reversed through activation of the rapamycin sensitive system and dephosphorylation of eukaryotic elongation factor 2. PMID- 15341531 TI - Involvement of p42/p44 MAPK, p38 MAPK, JNK and nuclear factor-kappa B in interleukin-1beta-induced matrix metalloproteinase-9 expression in rat brain astrocytes. AB - Matrix metalloproteinase (MMP)-9 expression induced by interleukin-1beta (IL 1beta) was investigated in rat brain astrocyte-1 (RBA-1). Here we report that the mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-kappaB) pathways participate in the induction of MMP-9 expression by IL-1beta. Zymographic, western blotting, and RT-PCR analyses showed that IL-1beta increased expression of MMP-9 mRNA and protein, which were inhibited by inhibitors of MEK1/2 (U0126), p38 (SB202190), and JNK (SP600125). In accordance with these findings, IL-1beta stimulated phosphorylation of p42/p44 MAPK, p38, and c-Jun N terminal kinase (JNK), which was attenuated by U0126, SB202190, or SP600125, respectively. Furthermore, this up-regulation of MMP-9 mRNA and protein was blocked by a specific NF-kappaB inhibitor helenalin. Consistently, IL-1beta stimulated translocation of NF-kappaB into the nucleus and degradation of inhibitory kappa B-alpha (IkappaB-alpha) was revealed by western blotting and immunofluorescence staining, which was blocked by helenalin, but not by U0126, SB202190, or SP600125. Taken together, these results suggest that in RBA-1 cells, activation of p42/p44 MAPK, p38, JNK and NF-kappaB pathways is essential for IL 1beta-induced MMP-9 gene expression via transcription and translation processes. An increased understanding of the signal transduction pathways involved in IL 1beta-induced MMP-9 expression on RBA-1 may be of potential therapeutic value in the treatment of inflammatory disease. PMID- 15341532 TI - Acetylcholinesterase inhibitors increase ADAM10 activity by promoting its trafficking in neuroblastoma cell lines. AB - Acetylcholinesterase inhibitors (AChEIs) are the only currently available drugs for treating Alzheimer's Disease (AD). Some authors have suggested a function of AChEIs not only in the induction of AChE overproduction and alternative splicing shifts but also a possible role of these drugs in amyloid metabolism beyond their well-known symptomatic effect. Here, we investigate the mechanisms of action of the AChEI donepezil on APP (amyloid precursor protein) metabolism and on the activity/trafficking of the alpha-secretase candidate ADAM 10, in differentiated human neuroblastoma cells (SH-SY5Y). In these cells, the activity of AChE is significantly decreased after 2 h of donepezil treatment. Further, SH-SY5Y cells released significantly more sAPPalpha into the medium, whereas total APP levels in cell lysates were unchanged. Interestingly, treated cells showed increased ADAM 10 levels in membrane compartments. This effect was prevented by pretreatment with tunicamycin or brefeldin, suggesting that donepezil affects trafficking and/or maturation of ADAM 10; additionally, this pretreatment significantly decreased sAPPalpha levels. Pre-incubation with atropine decreased release of sAPPalpha significantly but did not revert ADAM 10 activity to control levels further suggesting that donepezil acts not solely through a purely receptor mediated pathway. These findings indicate that donepezil exerts multiple mechanisms involving processing and trafficking of key proteins involved in AD pathogenesis. PMID- 15341533 TI - Potentiation of myelin proteolipid protein (Plp) gene expression is mediated through AP-1-like binding sites. AB - The myelin proteolipid protein (Plp) gene is expressed in oligodendrocytes and encodes the most abundant protein found in mature CNS myelin. Expression of the gene is dynamic and peaks during the active myelination period of CNS development. The surge in Plp gene activity during this period has been purported to be mediated by a positive regulatory region located within the first intron. This region, designated ASE for antisilencer/enhancer, is located approximately 1 kb downstream of exon 1 sequences and encompasses nearly 100 bp. However, neither the critical nucleotides within this region, nor the associated DNA-binding proteins have been identified. In the present study, DNase I footprinting analysis demonstrated widespread protection of the region on both the coding and non-coding strands suggesting that multiple transcription factors are likely involved. Targeting of putative DNA-protein binding sites contained within the ASE by gel shift, transfection and mutagenesis studies revealed the importance of several AP-1-like binding sites in governing high levels of Plp gene expression in oligodendrocytes. Our results suggest that factors, which bind to these sites, form the core of a multiprotein complex that assembles on the ASE and ultimately affects the temporal regulation of the gene in oligodendrocytes. PMID- 15341534 TI - Application of proteasomal inhibitors to mouse sympathetic neurons activates the intrinsic apoptotic pathway. AB - Proteasomal dysfunction may play a role in a number of neurodegenerative conditions, and in particular Parkinson's disease (PD) and related Lewy body (LB) diseases. Application of proteasomal inhibitors to neuronal cell culture systems is associated with survival-promoting effects or with cell death depending on the model system. We have applied pharmacological proteasomal inhibitors to cultured neonatal mouse sympathetic neurons in order to investigate whether these catecholaminergic neurons, which are affected in PD, are sensitive to proteasomal inhibition and, if so, which cell death pathway is activated. We report here that proteasomal inhibition leads to apoptotic death of mouse sympathetic neurons. This death is accompanied by caspase 3 activation and cytochrome c release from the mitochondria and is abrogated by caspase inhibition. Bax deletion prevented both cytochrome c release and caspase 3 activation, and also provided complete protection against proteasomal inhibition-induced death. Bcl-2 overexpression achieved a similar survival-promoting effect. There was no change in Bax levels following proteasomal inhibition, suggesting that Bax itself is not regulated by the proteasome in this cell culture system, and that a primary increase in Bax is unlikely to account for death. In contrast, levels of the BH3-only protein, Bim, increased with proteasomal inhibition. We conclude that proteasomal inhibition of mouse sympathetic neurons activates the intrinsic apoptotic pathway involving bcl 2 family members and the mitochondria. PMID- 15341535 TI - Neuregulin induces GABAA receptor beta2 subunit expression in cultured rat cerebellar granule neurons by activating multiple signaling pathways. AB - The GABAA receptor beta subunit is required to confer sensitivity to gamma aminobutyric acid (GABA), the major inhibitory neurotransmitter in the CNS. In previous studies we demonstrated that the growth and differentiation factor neuregulin 1 (NRG1) selectively induced expression of the beta2 subunit mRNA and encoded protein in rat cerebellar granule neurons in culture. In the present report we examine the signaling pathways that mediate this effect. These studies demonstrate that the effects of NRG1 on beta2 subunit polypeptide expression require activation of the ErbB4 receptor tyrosine kinase; its effects are inhibited by pharmacological blockade of ErbB4 phosphorylation or reduction of receptor level with an antisense oligodeoxynucleotide. The NRG1-induced activation of ErbB4 stimulates the mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K) and cyclin-dependent kinase-5 (cdk5) pathways. Pharmacological blockade of any of these pathways inhibits increased beta2 subunit expression, demonstrating that all three pathways are required to mediate the effects of NRG1 on GABAA receptor subunit expression in cerebellar granule neurons. These studies provide novel information concerning the actions of NRG1 on GABAA receptor expression in the CNS. PMID- 15341536 TI - Fetal alcohol exposure alters neurosteroid levels in the developing rat brain. AB - Neurosteroids are modulators of neuronal function that may play important roles in brain maturation. We determined whether chronic prenatal ethanol exposure altered neurosteroid levels in the developing brain. Rat dams were exposed to: (i) a 5% ethanol-containing liquid diet that produces peak maternal blood alcohol levels near the legal intoxication limit (approximately 0.08 g/dL); (ii) an isocaloric liquid diet containing maltose-dextrin instead of ethanol with pair feeding; (iii) rat chow ad libitum. Neurosteroid levels were assessed in offspring brains using radioimmunoassay or gas chromatography-mass spectrometry techniques. A prenatal ethanol exposure-induced increase in pregnenolone sulfate levels, but not dehydroepiandrosterone sulfate levels, was evident at the earliest time point studied (embryonic day 14). This effect lasted until post natal day 5. Levels of other neurosteroids were assessed at embryonic day 20; pregnenolone levels, but not allopregnanolone levels, were elevated. Pregnenolone sulfate levels were not altered in the maternal brain. Neither pregnenolone nor pregnenolone sulfate levels were significantly altered in the fetal liver, placenta and maternal blood, indicating that the effect of ethanol is not secondary to accumulation of peripherally-produced steroids. Fetal ethanol exposure has been shown to decrease both cellular and behavioral responsiveness to neurosteroids, and our findings provide a plausible explanation for this effect. PMID- 15341540 TI - A randomized, controlled trial of comprehensive geriatric assessment and multidisciplinary intervention after discharge of elderly from the emergency department--the DEED II study. AB - OBJECTIVES: To study the effects of comprehensive geriatric assessment (CGA) and multidisciplinary intervention on elderly patients sent home from the emergency department (ED). DESIGN: Prospective, randomized, controlled trial with 18 months of follow-up. SETTING: Large medical school-affiliated public hospital in an urban setting in Sydney, Australia. PARTICIPANTS: A total of 739 patients aged 75 and older discharged home from the ED were randomized into two groups. INTERVENTION: Patients randomized to the treatment group underwent initial CGA and were followed at home for up to 28 days by a hospital-based multidisciplinary outreach team. The team implemented or coordinated recommendations. The control group received usual care. MEASUREMENTS: The primary outcome measure was all admissions, to the hospital within 30 days of the initial ED visit. Secondary outcome measures were elective and emergency admissions, and nursing home admissions and mortality. Additional outcomes included physical function (Barthel Index (total possible score=20) and instrumental activities of daily living (/12) and cognitive function (mental status questionnaire (/10)). RESULTS: Intervention patients had a lower rate of all admissions to the hospital during the first 30 days after the initial ED visit (16.5% vs 22.2%; P=.048), a lower rate of emergency admissions during the 18-month follow-up (44.4% vs 54.3%; P=.007), and longer time to first emergency admission (382 vs 348 days; P=.011). There was no difference in admission to nursing homes or mortality. Patients randomized to the intervention group maintained a greater degree of physical and mental function (Barthel Index change from baseline at 6 months: -0.25 vs -0.75; P<.001; mental status questionnaire change from baseline at 12 months: -0.21 vs -0.64; P<.001). CONCLUSION: CGA and multidisciplinary intervention can improve health outcomes of older people at risk of deteriorating health and admission to hospital. Patients aged 75 and older should be referred for CGA after an ED visit. PMID- 15341541 TI - Use of the Physician Orders for Life-Sustaining Treatment program in Oregon nursing facilities: beyond resuscitation status. AB - OBJECTIVES: Program was designed to communicate resident/surrogate treatment preferences in the form of medical orders. To assess statewide nursing facility use of the Physician Orders for Life-Sustaining Treatment (POLST) and to identify the patterns of orders documented on residents' POLST forms. DESIGN: Telephone survey; on-site POLST form review. SETTING: Oregon nursing facilities. PARTICIPANTS: One hundred forty-six nursing facilities in the telephone survey; 356 nursing facility residents aged 65 and older at seven nursing facilities in the POLST form review. MEASUREMENTS: A telephone survey; onsite POLST form reviews. RESULTS: In the telephone survey, 71% of facilities reported using the POLST program for at least half of their residents. In the POLST form review, do not-resuscitate (DNR) orders were present on 88% of POLST forms. On forms indicating DNR, 77% reflected preferences for more than the lowest level of treatment in at least one other category. On POLST forms indicating orders to resuscitate, 47% reflected preferences for less than the highest level of treatment in at least one other category. The oldest old (> or = 85, n=167) were more likely than the young old (65-74, n=48) to have orders to limit resuscitation, medical treatment, and artificial nutrition and hydration. CONCLUSION: The POLST program is widely used in Oregon nursing facilities. A majority of individuals with DNR orders requested some other form of life extending treatment, and advanced age was associated with orders to limit treatments. PMID- 15341542 TI - The Physician Orders for Life-Sustaining Treatment program: Oregon emergency medical technicians' practical experiences and attitudes. AB - OBJECTIVES: To evaluate emergency medical technicians' (EMTs) experiences with the Physician Orders for Life-Sustaining Treatment (POLST) program and learn about attitudes regarding its effectiveness. DESIGN: Anonymous survey mailed to a stratified random sample. SETTING: Tri-County Portland, Oregon, area. PARTICIPANTS: A total of 572 Oregon EMT respondents (out of 1,048 surveys) were included in the analysis. MEASUREMENTS: Survey questions about experiences with the POLST form and opinions about POLST. RESULTS: Respondents were mostly male (76%) and paramedics (66%). Most respondents (73%) had treated a patient with a POLST, and 74% reported receiving education about POLST. EMTs reported that POLST, when present, changed treatment in 45% of cases. Seventy-five percent of the respondents agreed that the POLST form provides clear instructions about patient preferences, and 93% agreed that the POLST form is useful in determining which treatments to provide when the patient is in cardiopulmonary arrest. Fewer (63%) agreed that the form is useful in determining treatments when the patient has a pulse and is breathing. CONCLUSION: Most respondents have experience with the POLST program. EMTs find the POLST form useful and often use it to change treatment decisions for patients. PMID- 15341543 TI - Risk factors for harm in cognitively impaired seniors who live alone: a prospective study. AB - OBJECTIVES: To identify risk factors for harm due to self-neglect or behaviors related to disorientation in cognitively impaired seniors who live alone that can be used in primary care. DESIGN: Inception cohort followed prospectively for 18 months. SETTING: Participants were referred by their primary care physicians and community service agencies or were patients of several medical units of a large teaching hospital. PARTICIPANTS: One hundred thirty-nine community-residing participants, aged 65 and older who scored less than 131 on the Dementia Rating Scale and lived alone. MEASUREMENTS: Baseline Mini-Mental State Examination (MMSE); a social resources questionnaire; presence of chronic obstructive pulmonary disease (COPD), cerebrovascular disease, diabetes mellitus, Charlson Comorbidity Index, and medication use were examined as predictors of incident harm. Informants and primary care physicians provided information about the nature of any harm at 3-month intervals over an 18-month period. An incident of harm was included if it occurred as the result of self-neglect or behaviors related to disorientation, resulted in physical injury or property loss or damage, and required emergency community interventions. RESULTS: Based on the consensual agreement of four raters, 21.6% had an incident of harm. The proportional hazards model was highly significant (P<.001) and supported by bootstrapping estimates. Four variables were significantly predictive of time to incident harm: perception of fewer social resources, poorer performance on MMSE, presence of COPD, and presence of cerebrovascular disorders. CONCLUSION: Predictors of incident harm can be identified in the primary care setting and provide direction for the early identification of those at highest risk. Validation of findings with a new cohort is necessary. PMID- 15341544 TI - Effect of vascular lesions on cognition in Alzheimer's disease: a community-based study. AB - OBJECTIVES: To investigate whether clinical and neuropathological differences exist between Alzheimer's disease (AD) cases with and without vascular lesions neuropathologically diagnosed using Consortium to Establish a Registry for Alzheimer's Disease (CERAD) criteria. DESIGN: Descriptive observational study. SETTING: A community-based registry that identified incident dementia cases. PARTICIPANTS: Of the 124 subjects with available clinical and neuropathological assessments, 30 had AD lesions alone, and 18 had AD with vascular lesions. Patients with other neuropathological findings were excluded. MEASUREMENTS: Dependent measures included demographic, clinical, and neuropathological characteristics. Neuropathological diagnoses were made using the CERAD criteria and Braak and Braak staging. RESULTS: Of the 124 autopsied cases, 85 cases were diagnosed with neuropathological AD. Of these, 30 had pathology consistent with "pure" AD, whereas 18 had AD pathology with significant vascular lesions (AD/V). There were no differences in age, sex, or education between groups. AD/V cases had higher baseline and final Mini-Mental State Examination (MMSE) scores than pure AD cases, but after adjusting for education, differences in MMSE scores were not statistically significant. The AD/V group had significantly lower Braak staging than the pure AD group, after adjusting for education and final MMSE scores. CONCLUSION: In this comparison study of AD cases with and without vascular lesions, AD/V cases had less severe AD pathology than those with AD alone, indicating that cerebrovascular disease likely contributes to the severity of cognitive impairment in those with AD. Controlling for vascular risk factors in patients with AD may have a significant effect on severity of dementia. PMID- 15341545 TI - The effect of risperidone on nursing burden associated with caring for patients with dementia. AB - OBJECTIVES: To compare risperidone treatment with placebo for the burden to nursing staff of older nursing-home residents with dementia. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Long-term care institutions in Australia and New Zealand. PARTICIPANTS: Two hundred seventy-nine residents had evaluable data for comparison of oral risperidone with placebo treatment at 4 weeks, 8 weeks, and endpoint across the 12-week trial. MEASUREMENTS: The Modified Nursing Care Assessment Scale (M-NCAS) is a 28-item instrument that measures behavior occurrence in patients (patient outcome) and difficulty of those behaviors for nursing staff as rated by nurse carers (nurse outcome). Treatment groups were compared via analysis of covariance (ANCOVA). To aid score interpretation, M-NCAS effect sizes were examined by response category, with response defined as subjects with mean Cohen-Mansfield Agitation Inventory total aggression subscale score change of 4 points or greater, a clinical outcome for patients. RESULTS: Between-group ANCOVAs showed statistically significant reductions in nurse-rated M-NCAS scores from baseline to endpoint for the patients randomized to risperidone relative to placebo (P<.05). Effects were evident at 4 weeks and maintained over time. Effect sizes were moderate to large for responders and near zero for nonresponders. CONCLUSION: Risperidone treatment was associated with an early and sustained reduction in nursing staff burden as measured using the M-NCAS compared with placebo treatment for older nursing home residents with dementia. PMID- 15341546 TI - Cognitive decline in high-functioning older persons is associated with an increased risk of hospitalization. AB - OBJECTIVES: To examine hospital use for patients with evidence of cognitive decline indicative of early cognitive impairment. DESIGN: Medicare Part A hospital utilization data were linked to data from the MacArthur Research Network on Successful Aging Community Study to examine the association between baseline cognition and decline in cognitive function over a 3-year period and any hospitalization over that same period. SETTING: New Haven, Connecticut, and East Boston, Massachusetts. PARTICIPANTS: Subjects (N=598) were from two sites of the MacArthur Research Network on Successful Aging Community Study, a 7-year cohort study of community-dwelling older persons with high physical and cognitive functioning. MEASUREMENTS: Multivariate logistic regression was used to determine the association between any hospitalization over 3 years (1988-91) as the outcome variable and baseline cognitive function and decline in cognition over 3 years as primary predictor variables. Decline was based upon repeated (1988 and 1991) measures of delayed verbal recall and the Short Portable Mental Status Questionnaire (SPMSQ). RESULTS: Of 598 subjects, 48 died between 1988 and 1991. No baseline (1988) delayed recall scores or change in recall scores (1988-91) were associated with hospitalization. Although 48.2% declined on verbal memory scores, decline was not associated with risk of hospitalization. Of 494 subjects with complete 3-year data, 31.2% declined at least one point on the SPMSQ, and 4.7% declined more than two points. Among individuals aged 75 and older at baseline, the adjusted odds ratio for hospitalization for those who declined more than 2 points compared with those who declined less was 7.8 (95% confidence interval=2.0-30.8). CONCLUSION: Although specific memory tests were not associated with hospitalization, high-functioning older persons who experienced decline in overall cognitive function were more likely to be hospitalized. Variation in baseline cognitive function in this high-functioning cohort did not affect hospitalization, but additional research is needed to evaluate associations with other healthcare costs. PMID- 15341547 TI - Effectiveness of a group outpatient visit model for chronically ill older health maintenance organization members: a 2-year randomized trial of the cooperative health care clinic. AB - OBJECTIVES: To compare the effectiveness of Cooperative Health Care Clinic ((CHCC) group outpatient model for chronically ill, older health maintenance organization (HMO) patients) with usual care. DESIGN: Two-year, randomized, controlled trial conducted with recruitment from February 1995 through July of 1996. SETTING: Nonprofit group model HMO. PARTICIPANTS: Two hundred ninety-four adults (145 intervention and 149 usual care), aged 60 and older (mean age 74.1) with 11 or more outpatient visits in the prior 18 months, one or more self reported chronic conditions, and expressed interest in participating in a group clinic. INTERVENTION: Monthly group meetings held by patients' primary care physicians. MEASUREMENT: Differences in clinic visits, inpatient admissions, emergency room visits, hospital outpatient services, professional services, home health, and skilled nursing facility admissions; measures of patient satisfaction, quality of life, self-efficacy, and activities of daily living (ADLs). RESULTS: Outpatient, pharmacy services, home health, and skilled nursing facility use did not differ between groups, but CHCC patients had fewer hospital admissions (P=.012), emergency visits (P=.008), and professional services (P=.005). CHCC patients' costs were $41.80 per member per month less than those of control patients. CHCC patients reported higher satisfaction with their primary care physician (P=.022), better quality of life (P=.002), and greater self-efficacy (P=.03). Health status and ADLs did not differ between groups. CONCLUSION: The CHCC model resulted in fewer hospitalizations and emergency visits, increased patient satisfaction, and self-efficacy, but no effect on outpatient use, health, or functional status. PMID- 15341548 TI - Group treatment improves trunk strength and psychological status in older women with vertebral fractures: results of a randomized, clinical trial. AB - OBJECTIVES: To assess whether group exercise and coping classes reduce physical and psychological impairments and functional disability in older women with prevalent vertebral fractures (VFs). DESIGN: Randomized, controlled trial (modified cross-over) with site as unit of assignment; testing at baseline and 3, 6, 9, and 12 months. SETTING: Nine North Carolina retirement communities. PARTICIPANTS: One hundred eighty-five postmenopausal Caucasian women (mean age 81), each with at least one VFs. INTERVENTION: The intervention group had 6 months of exercise (3 meetings weekly, 45 minutes each) and coping classes (2 meetings weekly, 45 minutes each) in Phase 1, followed by 6 months of self maintenance. The control group had 6 months of health education control intervention (1 meeting weekly, 45 minutes) in Phase 1, followed by the intervention described above. MEASUREMENTS: Change in trunk extension strength, change in pain with activities, and change in psychological symptoms. RESULTS: Between-group differences in the change in trunk extension strength (10.68 foot pounds, P<.001) and psychological symptoms (-0.08, P=.011) were significant for Phase 1. Changes in pain with activities did not differ between groups (-0.03, P=.64); there was no change in the pain endpoint. In Phase 2, controls showed significant changes in trunk strength (15.02 foot pounds, P<.001) and psychological symptoms (-0.11, P=.006) from baseline. Change in pain with activities was not significant (-0.03, P=.70). During self-maintenance, the intervention group did not worsen in psychological symptoms, but improved trunk extension strength was not maintained. CONCLUSION: Weak trunk extension strength and psychological symptoms associated with VFs can be improved in older women using group treatment, and psychological improvements are retained for at least 6 months. PMID- 15341549 TI - Long-term prediction of incident hip fracture risk in elderly white women: study of osteoporotic fractures. AB - OBJECTIVES: To identify independent risk factors for first hip fracture over 10 years of follow-up. DESIGN: Prospective cohort study. SETTING: Four U.S. clinical centers. PARTICIPANTS: A total of 6,787 women aged 66 and older in the Study of Osteoporotic Fractures. MEASUREMENTS: Total hip bone mineral density (BMD) using dual-energy x-ray absorptiometry and a comprehensive set of potential risk factors were collected. Incident hip fractures were identified prospectively and confirmed using radiographic report. RESULTS: Six hundred two women (8.9%) had a hip fracture during a mean +/- standard deviation (SD) follow-up of 10.1 +/- 3.2 years. Older age, previous self-reported fracture after age 50, maternal history of hip fracture after age 50, greater height at age 25, impaired cognition, slower walking speed, nulliparity, type II diabetes mellitus, Parkinson's disease, and depth perception each independently predicted a 1.17- to 1.83-fold increase in hip fracture risk, whereas each SD (0.13 g/cm2) decrease in hip BMD was independently associated with a 1.84-fold increase in risk. Lower body mass index also was associated with an increased risk of hip fracture, although lower hip BMD largely explained this association. CONCLUSION: Although hip BMD is strongly related to hip fracture risk in elderly white women, other clinical risk factors also are independent predictors of long-term risk and provide additional insight into the prevention of fracture in high-risk women. Clinicians should be alert to factors other than BMD that place older women at a high risk of hip fracture. PMID- 15341550 TI - The effectiveness of a community-based program for reducing the incidence of falls in the elderly: a randomized trial. AB - OBJECTIVES: To test whether Stepping On, a multifaceted community-based program using a small-group learning environment, is effective in reducing falls in at risk people living at home. DESIGN: A randomized trial with subjects followed for 14 months. SETTING: The interventions were conducted in community venues, with a follow-up home visit. PARTICIPANTS: Three hundred ten community residents aged 70 and older who had had a fall in the previous 12 months or were concerned about falling. INTERVENTION: The Stepping On program aims to improve fall self efficacy, encourage behavioral change, and reduce falls. Key aspects of the program are improving lower-limb balance and strength, improving home and community environmental and behavioral safety, encouraging regular visual screening, making adaptations to low vision, and encouraging medication review. Two-hour sessions were conducted weekly for 7 weeks, with a follow-up occupational therapy home visit. MEASUREMENTS: The primary outcome measure was falls, ascertained using a monthly calendar mailed by each participant. RESULTS: The intervention group experienced a 31% reduction in falls (relative risk (RR)=0.69, 95% confidence interval (CI)=0.50-0.96; P=.025). This was a clinically meaningful result demonstrating that the Stepping On program was effective for community-residing elderly people. Secondary analysis of subgroups showed that it was particularly effective for men (n=80; RR=0.32, 95% CI=0.17-0.59). CONCLUSION: The results of this study renew attention to the idea that cognitive-behavioral learning in a small-group environment can reduce falls. Stepping On offers a successful fall-prevention option. PMID- 15341551 TI - Footwear style and risk of falls in older adults. AB - OBJECTIVES: To determine how the risk of a fall in an older adult varies in relation to style of footwear worn. DESIGN: Nested case-control study. SETTING: Group Health Cooperative, a large health maintenance organization in Washington state. PARTICIPANTS: A total of 1,371 adults aged 65 and older were monitored for falls over a 2-year period; 327 qualifying fall cases were compared with 327 controls matched on age and sex. MEASUREMENTS: Standardized in-person examinations before fall occurrence, interviews about fall risk factors after the fall occurred, and direct examination of footwear were conducted. Questions for controls referred to the last time they engaged in an activity broadly similar to what the case was doing at the time of the fall. RESULTS: Athletic and canvas shoes (sneakers) were the styles of footwear associated with lowest risk of a fall. Going barefoot or in stocking feet was associated with sharply increased risk, even after controlling for measures of health status (adjusted odds ratio=11.2, 95% confidence interval (CI)=2.4-51.8). Relative to athletic/canvas shoes, other footwear was associated with a 1.3-fold increase in the risk of a fall (95% CI=0.9-1.9), varying somewhat by style. CONCLUSION: Contrary to findings from gait-laboratory studies, athletic shoes were associated with relatively low risk of a fall in older adults during everyday activities. Fall risk was markedly increased when participants were not wearing shoes. PMID- 15341552 TI - Changes in sexual function in middle-aged and older men: longitudinal data from the Massachusetts Male Aging Study. AB - OBJECTIVES: To describe within-individual change in sexual function over a 9-year period and to determine whether the amount of change differs by age group. DESIGN: Cohort study; participants interviewed at baseline (1987-89) and follow up (1995-97). SETTING: Population-based; communities surrounding Boston, Massachusetts. PARTICIPANTS: One thousand eighty-five men aged 40 to 70 at baseline (born between 1917 and 1947) with complete baseline and follow-up sexual function data. MEASUREMENTS: Within-person change (follow-up minus baseline) in the following sexual function variables: sexual intercourse, erection frequency, sexual desire, ejaculation with masturbation, satisfaction with sex, and difficulty with orgasm. RESULTS: Unadjusted analyses showed significant longitudinal changes over the 9-year period in all domains of sexual function except frequency of ejaculation with masturbation, which showed no change between baseline and follow-up. Adjusted for baseline sexual function, within-person change in all outcomes was strongly related to age, with decline in sexual function becoming more pronounced with increasing age. For example, over the 9 year study period, sexual intercourse or activity frequency decreased by less than once per month, two times per month, and three times per month in men in their 40s, 50s, and 60s, respectively. Number of erections per month declined by 3, 9, and 13 in men in their 40s, 50s, and 60s, respectively. CONCLUSION: This research fills a major gap in the literature by providing age-specific estimates of change in sexual functioning over a 9-year period in a cohort of unselected men. PMID- 15341553 TI - Alcohol screening results in elderly male veterans: association with health status and mortality. AB - OBJECTIVES: To evaluate the association between alcohol screening results and health status or mortality in elderly patients. DESIGN: Cross-sectional and longitudinal cohort studies. SETTING: Primary care clinics at seven Veterans Affairs medical centers. PARTICIPANTS: A total of 16,958 male patients aged 65 and older. MEASUREMENTS: Alcohol screening questionnaires were used to divide patients into four mutually exclusive groups: drinkers who screened negative or positive for problem drinking and nondrinkers (in the previous year) who screened negative or positive for problem drinking. A subset of patients (n=12,491) completed a measure of health status, the Medical Outcomes Study Short Form (SF 36). All-cause mortality was ascertained using Veterans Affairs data over a mean follow-up of 2.8 years. RESULTS: Forty-eight percent of participants reported drinking in the previous year. Half of the drinkers and nondrinkers screened positive for problem drinking. The SF-36 item scores were consistently higher for drinkers than for nondrinkers and for patients who screened negative for problems than for those who screened positive. Similarly, survival was consistently better in drinkers than nondrinkers (mortality hazard ratio (HR)=0.74, 95% confidence interval (CI)=0.68-0.82) and in patients who screened negative for problem drinking than in those who screened positive (mortality HR=0.87, 95% CI=0.80 0.95). Nondrinkers who screened positive for problem drinking had the poorest health status and survival, whereas drinkers who screened negative for problem drinking had the best health status and survival. CONCLUSION: Elderly patients who reported drinking alcohol and those who screened negative for problem drinking had consistently better health status and survival than those who did not drink and those who screened positive. PMID- 15341554 TI - A randomized, double-blind, placebo-controlled clinical trial of the Chinese herbal medicine "ba wei di huang wan" in the treatment of dementia. AB - OBJECTIVES: To evaluate whether a traditional Chinese herbal medicine, ba wei di huang wan (BDW), improves cognitive and physical functioning in dementia patients. DESIGN: An 8-week randomized, double-blind, placebo-controlled trial. SETTING: Long-term-care facility in Japan. PARTICIPANTS: Thirty-three patients with mild to severe dementia (7 men and 26 women; mean age +/- standard deviation=84.4 +/- 7.8) were recruited and enrolled from May 2002 through September 2002. INTERVENTION: Participants were randomly assigned to the active drug (BDW) group (n=16) or the placebo group (n=17) and treated for 8 weeks. MEASUREMENT: Cognitive function and activities of daily living (ADLs); palsatility index. RESULTS: After the trial, cognitive function as assessed using the Mini-Mental State Examination (MMSE) significantly improved from 13.5 +/- 8.5 to 16.3 +/- 7.7 (P<.01, 95% confidence interval (CI)=-4.1 to -1.4) in the BDW group. The ADL score in the Barthel Index also significantly changed, from 61.8 +/- 34.6 to 78.9 +/- 21.1 (P<.01, 95% CI=-26.2 to -7.9). In contrast, MMSE and Barthel Index scores of the placebo group showed no significant change. Eight weeks after the end of the administration, MMSE and Barthel Index scores of the BDW group declined to the baseline level. The pulsatility index in the internal carotid artery as measured using Doppler sonography significantly decreased in the BDW group (2.5 +/- 1.7 to 1.9 +/- 0.5, P<.05) but not in the placebo group. CONCLUSION: These results argue the benefits of BDW in the treatment of dementia. PMID- 15341555 TI - Fall-risk assessment and management in clinical practice: views from healthcare providers. AB - OBJECTIVES: To determine the extent to which healthcare providers reportedly address evidence-based fall risk factors in older patients after exposure to an educational intervention and to determine barriers reportedly encountered when these healthcare providers intervene with or refer older patients with identified fall-risk factors. DESIGN: Cross-sectional study using a structured interview. SETTING: Geographic area of Connecticut where the Connecticut Collaboration for Fall Prevention (CCFP) has been implemented. PARTICIPANTS: Emergency department (ED) physicians, hospital-based discharge planners or care coordinators (nurses or social workers), home health agency nurses, and office-based primary care physicians (total n=33) after exposure to the CCFP implementation team. MEASUREMENTS: Self-reported practices (direct intervention or referral) and barriers when addressing seven evidence-based risk factors for falls: gait and transfer impairments, balance disturbances, multiple medications, postural hypotension, sensory and perceptive deficits, foot and footwear problems, and environmental hazards. RESULTS: Respondents were most likely to report directly intervening with or referring older patients for gait and transfer impairments (85%) and balance disturbances (82%) and least likely to do so when encountering foot or footwear problems (58%) and sensory or perceptive deficits (61%). ED physicians reported lowest rates of direct intervention or referral for foot or footwear problems (20%), home health agency nurses for sensory or perceptive deficits (50%), and office-based primary care physicians for foot or footwear problems (50%). Patient compliance was the most commonly reported barrier to successful direct intervention across several risk factors, whereas inadequate availability of other healthcare providers and lack of Medicare reimbursement were the most commonly reported barriers to successful patient referrals. CONCLUSION: After exposure to the CCFP implementation team, the majority of healthcare providers reported directly intervening or referring patients when addressing all risk factors, but results pinpointed specific healthcare provider groups with room for improvement in assessment and management of specific risk factors. Patient education appears to be a necessary adjunct to healthcare provider training, because patient compliance was a reported barrier to optimal intervention by healthcare providers. PMID- 15341556 TI - Detection and management of falls and instability in vulnerable elders by community physicians. AB - OBJECTIVES: To investigate quality of care for falls and instability provided to vulnerable elders. DESIGN: Six process of care quality indicators (QIs) for falls and instability were developed and applied to community-living persons aged 65 and older who were at increased risk of death or decline. QIs were implemented using medical records and patient interviews. SETTING: Northeastern and southwestern United States. PARTICIPANTS: Three hundred seventy-two vulnerable elders enrolled in two senior managed care plans. MEASUREMENTS: Percentage of QIs satisfied concerning falls or mobility disorders. RESULTS: Of the 372 consenting vulnerable elders with complete medical records, 57 had documentation of 69 episodes of two or more falls or fall with injury during the 13-month study period (14% of patients fell per year, 18% incidence). Double this frequency was reported at interview. An additional 22 patients had documented mobility problems. Clinical history of fall circumstances, comorbidity, medications, and mobility was documented from 47% of fallers and two or more of these four elements from 85%. Documented physical examination was less complete, with only 6% of fallers examined for orthostatic blood pressure, 7% for gait or balance, 25% for vision, and 28% for neurological findings. The evaluation led to specific recommendations in only 26% of cases, but when present they usually led to appropriate treatment modalities. Mobility problems without falls were evaluated with gait or balance examination in 23% of cases and neurological examination in 55%. CONCLUSION: Community physicians appear to underdetect falls and gait disorders. Detected falls often receive inadequate evaluation, leading to a paucity of recommendations and treatments. Adhering to guidelines may improve outcomes in community-dwelling older adults. PMID- 15341557 TI - A comparison of gait characteristics between older women with and without peripheral neuropathy in standard and challenging environments. AB - OBJECTIVES: To compare gait patterns in older women with and without peripheral neuropathy (PN) in standard (smooth surface, normal lighting) and challenging environments (CE) (irregular surface, low lighting). DESIGN: Observational, controlled study of 24 subjects. SETTING: Biomechanical research laboratory. PARTICIPANTS: Twenty-four older women, 12 with PN and 12 without PN (mean age +/- standard deviation =67.1 +/- 7.9 and 70.2 +/- 4.3, respectively). MEASUREMENTS: Gait parameters and, in the 12 PN subjects, neuropathy severity. RESULTS: The CE was associated with increases in step width, step-width variability, step-width range, step width-to-step length ratio, step time and step-time variability, and decreases in step length and speed. The PN subjects demonstrated a greater step width-to-step length ratio and step time and shorter step length and slower speed than the control subjects. In adapting to the CE, the PN subjects demonstrated greater increases in step width-to-step length ratio and step-time variability and a greater decrease in step length than did the control subjects. In the standard environment, only one gait parameter correlated with PN severity, whereas in the CE, four gait parameters did so. CONCLUSION: The subjects demonstrated a gait that was slower, less efficient, and more variable temporally and in the frontal plane in the CE. Control and PN subjects demonstrated similar variability in medial-lateral step placement in the CE but at the cost of speed and efficiency for the PN subjects. Because the CE magnified gait differences between the two groups of subjects and caused gait changes in the PN subjects that correlated with PN severity, the CE may offer improved resolution for detecting gait abnormalities. PMID- 15341558 TI - Blood pressure components and cardiovascular events in older adults: the Rotterdam study. AB - OBJECTIVES: To compare the strength of the relative risks of systolic (SBP) diastolic blood pressure (DBP) and pulse pressure (PP) as predictors of myocardial infarction and stroke in older adults. DESIGN: Prospective cohort study. SETTING: The Rotterdam Study, a Dutch population-based study. PARTICIPANTS: A total of 4,234 subjects aged 55 and older with no previous myocardial infarction (MI) or stroke at baseline. MEASUREMENTS: Blood pressure levels at baseline, first MI and stroke, all-cause mortality during follow-up. RESULTS: During follow-up, 205 subjects had an MI (average follow-up period 7 years), 137 subjects had a stroke (average follow-up period 6.1 years), and 748 subjects died. A 1-standard deviation difference in SBP, DBP, and PP was associated with relative risks of MI of 1.24 (95% confidence interval (CI)=1.06 1.46), 1.07 (0.92-1.25), and 1.25 (1.07-1.48), respectively. Corresponding relative risks for stroke were 1.59 (1.37-1.86), 1.27 (1.10-1.48), and 1.48 (1.27 1.72). For all-cause mortality the corresponding relative risks and 95% CI were 1.21 (1.11-1.31), 1.06 (0.99-1.14), and 1.20 (1.10-1.31). CONCLUSION: The results of this study suggest that, in a population of apparently healthy older adults, PP is not a better predictor of cardiovascular events and all-cause mortality than SBP. PMID- 15341559 TI - Comparison of fracture, cardiovascular event, and breast cancer rates at 3 years in postmenopausal women with osteoporosis. AB - OBJECTIVES: To compare event rates for osteoporotic fractures, cardiovascular events, and breast cancer in postmenopausal women with osteoporosis. DESIGN: A prospective, observational study of the placebo group in the double-blind, randomized Multiple Outcomes of Raloxifene Evaluation trial. SETTING: One hundred eighty clinical research centers in 25 countries. PARTICIPANTS: Postmenopausal women (n=2,565, mean age=67) with osteoporosis were given calcium (500 mg/d) and vitamin D (400-600 IU/d) supplements. MEASUREMENTS: The occurrence of at least one new fracture, cardiovascular event, or breast cancer diagnosis at 3 years was identified and adjudicated. RESULTS: The occurrence of any fracture was the most common event in these women. In women without prevalent vertebral fractures (n=1,627), the event rates per 1,000 patient-years were 45.4 for any fracture, 15.2 for vertebral fracture, 4.7 for clinical vertebral fracture, 0.9 for hip fracture, 8.3 for any cardiovascular event, and 5.2 for all breast cancer. In women with prevalent vertebral fractures (n=938), the event rates per 1,000 patient-years were 117.4 for any new fracture, 77.1 for new vertebral fracture, 25.7 for clinical vertebral fracture, 5.8 for hip fracture, 15.1 for any cardiovascular event, and 2.6 for all breast cancer. The effect of prevalent fracture status on event rates was not dependent on whether women were older or younger than 65, but women aged 65 and older had a 3.6 times greater occurrence of cardiovascular events than younger women, irrespective of prevalent fracture status. CONCLUSION: These data on the relative incidence of clinically significant skeletal and extra-skeletal outcomes may be useful in choosing an agent for health maintenance for postmenopausal women with osteoporosis. PMID- 15341560 TI - Older women and HIV: how much do they know and where are they getting their information? AB - OBJECTIVES: To assess older urban women's knowledge about sexual transmission of human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) and to evaluate the relationship between their HIV/AIDS knowledge level and sources of information. DESIGN: Cross-sectional survey conducted between June 2001 and July 2002. Trained research assistants administered a questionnaire in a face-to face interview. SETTING: General medicine clinic in a large public hospital in a high HIV/AIDS incidence area. PARTICIPANTS: Five hundred fourteen women aged 50 and older. MEASUREMENTS: Nine questions assessing knowledge of risk of HIV sexual transmission with potential scores ranging from 0 to 9 correct answers. Participants identified all sources of HIV information. RESULTS: The mean knowledge score was 3.7 out of a possible 9 correct responses (range 0 (3%) to 8 (1%)). Younger age, employment, and higher educational level were associated with higher knowledge scores, whereas marital status was unrelated. No respondent correctly answered all of the nine questions. The most commonly identified sources of HIV/AIDS information were television (85%), friends (54%), and newspapers (51%). Only 38% of respondents identified health professionals as a source of information about HIV/AIDS. Health professionals, newspapers, and family members were each independently associated with higher knowledge scores (P<.05). CONCLUSION: Older women in a general medicine clinic had limited knowledge of sexual transmission of HIV. HIV/AIDS education specifically targeted to this subpopulation is warranted, and health professionals may have an important role in disseminating such messages. PMID- 15341561 TI - Validation of the Late-Life Function and Disability Instrument. AB - OBJECTIVES: To assess the concurrent and predictive validity of the Late-Life Function and Disability Instrument (LLFDI). DESIGN: Cross-sectional. SETTING: University-based human physiology laboratory. PARTICIPANTS: One hundred one men and women aged 80.8 +/- 0.4. MEASUREMENTS: A short physical performance battery (SPPB) and a self-paced 400-m walk (400-m W) were used as performance tests of lower extremity function. The LLFDI was used to assess self-reported function and physical disability. Partial correlations adjusted for age and body mass index were used to determine the concurrent and predictive validity of the LLFDI. Statistical significance was accepted at P<.004 using a testwise correction. RESULTS: LLFDI Overall Function scores were moderately associated with the SPPB (r=0.65, P<.001), 400-m W gait speed (r=0.69, P<.001), and measures of lower extremity function. Correlations of the two lower extremity subscores of the LLFDI (correlation coefficient (r)=0.63-0.73, P<.001) were greater than for the LLFDI upper extremity subscores (r=0.19-0.26, P>.004). Performance measures of function predicted disability limitations in the range of r=0.37-0.44 (P<.001) and disability frequency in the range of r=0.16-0.20 (P>.004). CONCLUSION: These findings support the concurrent and predictive validity of the LLFDI. Results support the use of the LLFDI scales as a substitute for physical performance tests when self-report is a preferred data-collection format. PMID- 15341562 TI - Clinical Global Impression of Change in Physical Frailty: development of a measure based on clinical judgment. AB - OBJECTIVES: To expand the ability to assess physical frailty by developing a Clinical Global Impression of Change in Physical Frailty (CGIC-PF) instrument. DESIGN: Qualitative and quantitative instrument development. SETTING: Academic centers. PARTICIPANT: s Six expert panel members, 46 clinicians, 24 patients, and 12 caregivers. MEASUREMENTS: Literature review and structured group processes with experts, clinicians, and consumers were used to generate an initial list of domains and indicators. Structured interviews with clinical experts in the area of frailty were used to establish relevance and feasibility of measurement of domains. Interrater reliability was assessed through a Web-based study. Geriatricians pilot tested the feasibility of the baseline CGIC-PF with 10 patients. RESULTS: The CGIC-PF includes six intrinsic domains (mobility, balance, strength, endurance, nutrition, and neuromotor performance) and seven consequences domains (medical complexity, healthcare utilization, appearance, self-perceived health, activities of daily living, emotional status, and social status). Each domain has two to four clinical indicators. Change is scored on a 7 point scale from markedly worse to markedly improved. Average interrater reliability of the CGIC-PF for the Web-based cases was 0.97. Geriatricians completed a baseline CGIC-PF on their own patients in 10 minutes or less. CONCLUSION: The CGIC-PF is a structured assessment of change in physical frailty with defined content and process. It has strong face validity, reliability, and feasibility for use in clinical research. It may be useful as one criterion of change and as an anchor for change in other measures. PMID- 15341563 TI - Navigating federalwide assurance requirements when conducting research in community-based care settings. AB - There is an urgent need for research on quality of life and healthcare delivery for older adults living in community-based care settings, yet implementing current federalwide assurance (FWA) requirements can be a challenge in these settings. This paper discusses FWA requirements for engagement in federally funded research as the requirements pertain to community-based care settings. Factors that impede community facilities in achieving FWA approval include lack of organizational structure to provide oversight for the ethical conduct of research, administrator concerns regarding potential liability associated with obtaining the FWA, lack of resources to complete required paperwork, and lack of staff knowledge about human subjects protection and federal requirements for participating in research. Effects of the FWA process on investigators include the burden of extra time needed to support community-based facilities to acquire a FWA and concerns that studies may be limited to only those community facilities with the resources to complete the FWA process. Investigator-initiated strategies for conducting research in community-based settings include considering study designs that are exempt from the FWA process and proactively assisting community based facilities to acquire FWA status. Investigators need to work with potential research sites and the office for human research protections to ensure that subjects are protected without shifting the burden of protection to ill-prepared community administrators. PMID- 15341565 TI - Oregon's lessons for improving advance care planning. PMID- 15341566 TI - Dementia patients who live alone: research and clinical challenges. PMID- 15341567 TI - Elixir of life. PMID- 15341569 TI - Inappropriately defining "inappropriate medication for the elderly". PMID- 15341570 TI - Criteria for prescribing require further study. PMID- 15341572 TI - Metabolic improvement and abdominal fat redistribution in Werner syndrome by pioglitazone. PMID- 15341573 TI - Treatment of fractures of the atrophic mandible in the elderly. PMID- 15341574 TI - Chronic and forgotten? PMID- 15341576 TI - Plasma levels of inflammatory C-reactive protein and interleukin-6 predict outcome in elderly patients with stroke. PMID- 15341577 TI - Stroke units and acute care for elders model of care. PMID- 15341578 TI - Protein malnutrition is adversely associated with swallowing recovery in tube-fed older people. PMID- 15341579 TI - Is adult definition of anemia applicable to a geriatric population? Study of erythrocyte parameters in Indian geriatric inpatients. PMID- 15341580 TI - Pesticide exposure and dementia. PMID- 15341581 TI - Mental functions and physical performance abilities as predictors of falling in a geriatric evaluation and rehabilitation unit. PMID- 15341582 TI - Arterial carboxyhemoglobin concentrations in elderly patients with operable non small cell lung cancer. PMID- 15341583 TI - Risus sardonicus still beyond the corner. PMID- 15341584 TI - From Nascher to now-75 years of U.S. geriatrics. PMID- 15341585 TI - His-story as a dimension of the present. PMID- 15341586 TI - Optical detection of neural function in the chick visual pathway in the early stages of embryogenesis. AB - We investigated the developmental pattern of functional synaptogenesis in the chick visual pathway using a multiple-site optical recording method. Responses to optic nerve stimulation were recorded from the diencephalon and mesencephalon of the chick embryo. The first excitatory postsynaptic responses to optic nerve stimulation appeared in the contralateral diencephalon at Hamburger-Hamilton stage 27, which corresponds to an incubation day 5.5 (E5.5). At more developed stages, the optical signals evoked by optic nerve stimulation spread to several different regions, including the tectum and extra-tectal visual nuclei. We constructed maps of neural activity in the diencephalon and mesencephalon at different stages to investigate the spatio-temporal patterns of functional development in the chick visual system. The maps revealed that distinct postsynaptic response areas in the extra-tectal regions showed different onsets of activity, suggesting that the corresponding visual nuclei exhibit different time courses of functional synaptogenesis. We also identified the onset and location of the first functional synaptic connection in the optic tectum, which had been a point of controversy in earlier studies. In the tectal region, the action potential and the excitatory postsynaptic potential first appeared at E8, although these signals were recognized in the tecto/tegmental region at E7. The response area expanded with retinotectal fibre elongation, and reached the area centralis at E9. These results show that the onset of synaptic function in the tectum occurs 2-3 days earlier than was previously reported. PMID- 15341587 TI - Fractalkine (CX3CL1) and fractalkine receptor (CX3CR1) distribution in spinal cord and dorsal root ganglia under basal and neuropathic pain conditions. AB - Fractalkine is a unique chemokine reported to be constitutively expressed by neurons. Its only receptor, CX3CR1, is expressed by microglia. Little is known about the expression of fractalkine and CX3CR1 in spinal cord. Given that peripheral nerve inflammation and/or injury gives rise to neuropathic pain, and neuropathic pain may be partially mediated by spinal cord glial activation and consequent glial proinflammatory cytokine release, there must be a signal released by affected neurons that triggers the activation of glia. We sought to determine whether there is anatomical evidence implicating spinal fractalkine as such a neuron-to-glia signal. We mapped the regional and cellular localization of fractalkine and CX3CR1 in the rat spinal cord and dorsal root ganglion, under basal conditions and following induction of neuropathic pain, employing both an inflammatory (sciatic inflammatory neuropathy; SIN) as well as a traumatic (chronic constriction injury; CCI) model. Fractalkine immunoreactivity and mRNA were observed in neurons, but not glia, in the rat spinal cord and dorsal root ganglia, and levels did not change following either CCI or SIN. By contrast, CX3CR1 was expressed by microglia in the basal state, and the microglial cellular concentration was up-regulated in a regionally specific manner in response to neuropathy. CX3CR1-expressing cells were identified as microglia by their cellular morphology and positive OX-42 and CD4 immunostaining. The cellular distribution of fractalkine and CX3CR1 in the spinal circuit associated with nociceptive transmission supports a potential role in the mechanisms that contribute to the exaggerated pain state in these models of neuropathy. PMID- 15341588 TI - Long-term changes in the molecular composition of the glial scar and progressive increase of serotoninergic fibre sprouting after hemisection of the mouse spinal cord. AB - The scarring process occurring after adult central nervous system injury and the subsequent increase in the expression of certain extracellular matrix molecules are known to contribute to the failure of axon regeneration. This study provides an immunohistochemical analysis of temporal changes (8 days to 1 year) in the cellular and molecular response of the Swiss mouse spinal cord to a dorsal hemisection and its correlation with the axonal growth properties of a descending pathway, the serotoninergic axons. In this lesion model, no cavity forms at the centre of the lesion. Instead, a dense fibronectin-positive tissue matrix occupies the centre of the lesion, surrounded by a glial scar mainly constituted by reactive astrocytes. NG2 proteoglycan and tenascin-C, potential axon growth inhibitors, are constantly associated with the central region. In the glial scar, tenascin-C is never observed and the expression of chondroitin sulphate proteoglycans (revealed with CS-56 and anti-NG2 antibodies) highly increases in the week following injury to progressively return to their control level. In parallel, there is an increasing expression of the polysialilated neural cell adhesion molecule by reactive astrocytes. These molecular changes are correlated with a sprouting process of serotoninergic axons in the glial scar, except in a small area in contact with the central region. All these observations suggest that while a part of the glial scar progressively becomes permissive to axon regeneration after mouse spinal cord injury, the border of the glial scar, in contact with the fibronectin-positive tissue matrix, is the real barrier to prevent axon regeneration. PMID- 15341589 TI - Dynamic changes of the anti- and pro-apoptotic proteins Bcl-w, Bcl-2, and Bax with Smac/Diablo mitochondrial release after photothrombotic ring stroke in rats. AB - The anti-apoptotic proteins Bcl-w and Bcl-2 and the pro-apoptotic protein Bax may mediate cell death or survival via regulation of the mitochondria including second mitochondria-derived activator of caspase (Smac)/direct inhibitor of apoptosis protein (IAP)-binding protein with low pI (DIABLO) release. This study aimed to explore alterations in Bcl-w, Bcl-2, and Bax and the relationship between these proteins and Smac/DIABLO by means of in situ hybridization, immunohistochemical (IHC) staining, and Western blots after low- and high intensity photothrombotic ring stroke. At 4 h after low-intensity irradiation, we found widespread bcl-w overexpression on both the mRNA and protein levels in the bilateral cortex except the ring lesion region and in subcortical regions. A prolonged elevation of Bcl-2 with relatively unchanged Bax in the mitochondrial fraction was demonstrated from 4 to 72 h. These upregulated anti-apoptotic proteins combined with little Smac/DIABLO release might be associated with increased cell survival and thereby remarkable morphological recovery after low intensity irradiation. After high-intensity irradiation, we observed decreased bcl-w and bcl-2 mRNA with increased Bcl-2 protein in the cytosolic fraction, whereas the Bax protein remained in scattered ischaemic cells in the ring lesion and the region at risk that corresponded with release of Smac/DIABLO from mitochondria to the cytosol at 1-24 h. These changes might be related to the massive cell death observed after high-intensity irradiation. Taken together, the balance and the location of anti-apoptotic proteins vs. pro-apoptotic proteins could be associated with the translocation of Smac/DIABLO from the mitochondria to the cytosol and therefore closely related to cell death or survival after focal cerebral ischaemia. PMID- 15341590 TI - BDNF increases rat brain mitochondrial respiratory coupling at complex I, but not complex II. AB - Brain-derived neurotrophic factor (BDNF) governs both the selective survival of neurons during development and the experience-based regulation of synaptic strength throughout life. BDNF produced a concentration-dependent increase in the respiratory control index (RCI, a measure of the efficiency of respiratory coupling, ATP synthesis and organelle integrity) of rat brain mitochondria. This effect was mediated via a MAP kinase pathway and highly specific for oxidation of glutamate plus malate (complex I) by brain mitochondria. The oxidation by brain mitochondria of the complex II substrate succinate was unaffected by BDNF. The failure of BDNF to modify respiratory activity associated with mitochondrial preparations isolated from rat liver indicates that the actions of the neurotrophin are tissue specific. BDNF also increased the RCI values associated with Ca2+ -induced respiration to a similar extent. This is the first demonstration that BDNF, in addition to modifying neuronal plasticity, can modify brain metabolism and the efficiency of oxygen utilization. The finding that neurotrophins can alter mitochondrial oxidative efficiency has important implications for neurodegenerative and psychiatric diseases. PMID- 15341591 TI - Deletion of the adenosine A1 receptor gene does not alter neuronal damage following ischaemia in vivo or in vitro. AB - Extracellular adenosine is dramatically increased during cerebral ischaemia and is considered to be neuroprotective due to its inhibitory effect on synaptic transmission mediated by the adenosine A1 receptor (A1R). We investigated the importance of the A1R in a mouse model of global ischaemia and in a murine hippocampal slice culture model of in vitro ischaemia, using mice with the A1R gene deleted. In brains from mice lacking the A1R, damage induced by global ischaemia was similar to that in wild-type animals. In contrast, treatment with a selective A1R antagonist [8-cyclo-pentyl theophylline (8-CPT)], administered before the ischaemic insult in naive wild-type mice, exacerbated the neuronal damage following global ischaemia. Although the inhibitory action of adenosine on excitatory neurotransmission in hippocampal slices was lost in A1R knockout mice, there was no difference in damage between slices from wild-type and knockout mice after in vitro ischaemia. The results suggest that some effects of the A1R are compensated for in knockout animals. PMID- 15341592 TI - Integration and differentiation of neural stem cells after transplantation into the dysmyelinated central nervous system of adult mice. AB - Mutant mice deficient in the myelin-associated glycoprotein (MAG) and the nonreceptor-type tyrosine kinase Fyn are characterized by a severely hypomyelinated central nervous system (CNS) and morphologically abnormal myelin sheaths. Despite this pronounced phenotype, MAG/Fyn-deficient mice have a normal longevity. In the present study, we took advantage of the normal life expectancy of this myelin mutant and grafted neural stem cells (NSCs) into the CNS of MAG/Fyn-deficient mice to study in short- and long-term experiments the fate of NSCs in adult dysmyelinated brains. Neural stem cells were isolated from spinal cords of transgenic mouse embryos ubiquitously expressing enhanced green fluorescent protein. Cells were expanded in vitro in the presence of mitogens for up to 5 weeks before they were grafted into the lateral ventricles or injected into white matter tracts. Analysis of mutant brains 3-15 weeks after intracerebroventricular transplantation of NSCs revealed only limited integration of donor cells into the host brains. However, injection of NSCs directly into white matter tracts resulted in widespread distribution of donor cells within the host tissue. Donor cells survived for at least 15 weeks in adult host brains. The majority of grafted cells populated white matter tracts and differentiated into oligodendrocytes that myelinated host axons. Results suggest that intraparenchymal transplantation of NSCs might be a strategy to reconstruct myelin in dysmyelinated adult brains. PMID- 15341593 TI - Functional repair after dorsal root rhizotomy using nerve conduits and neurotrophic molecules. AB - Functional recovery after large excision of dorsal roots is absent because of both the limited regeneration capacity of the transected root, and the inability of regenerating sensory fibers to traverse the dorsal root entry zone. In this study, bioresorbable guidance conduits were used to repair 6-mm dorsal root lesion gaps in rats, while neurotrophin-encoding adenoviruses were used to elicit regeneration into the spinal cord. Polyester conduits with or without microfilament bundles were implanted between the transected ends of lumbar dorsal roots. Four weeks later, adenoviruses encoding NGF or GFP were injected into the spinal cord along the entry zone of the damaged dorsal roots. Eight weeks after injury, nerve regeneration was observed through both types of implants, but those containing microfilaments supported more robust regeneration of calcitonin gene related peptide (CGRP)-positive nociceptive axons. NGF overexpression induced extensive regeneration of CGRP(+) fibers into the spinal cord from implants showing nerve repair. Animals that received conduits containing microfilaments combined with spinal NGF virus injections showed the greatest recovery in nociceptive function, approaching a normal level by 7-8 weeks. This recovery was reversed by recutting the dorsal root through the centre of the conduit, demonstrating that regeneration through the implant, and not sprouting of intact spinal fibers, restored sensory function. This study demonstrates that a combination of PNS guidance conduits and CNS neurotrophin therapy can promote regeneration and restoration of sensory function after severe dorsal root injury. PMID- 15341594 TI - Effects of muscarinic acetylcholine receptor activation on membrane currents and intracellular messengers in medium spiny neurones of the rat striatum. AB - Acetylcholine, acting through muscarinic receptors, modulates the excitability of striatal medium spiny neurones. However, the underlying membrane conductances and intracellular signalling pathways have not been fully determined. Our aim was to characterize excitatory effects mediated by M1 muscarinic acetylcholine receptors in these neurones using whole-cell patch-clamp recordings in brain slices of postnatal rats. Under voltage-clamp, muscarine evoked an inward current associated with an increase in cell membrane resistance. The current, which reversed at -85 mV, was sensitive to the M1 receptor antagonist pirenzepine. Blocking the potassium conductance attenuated the response and the residual current was further reduced by ruthenium red (50 microm) and reversed at +15 mV. Simultaneous recordings from cholinergic interneurones and medium spiny neurones in conjunction with spike-triggered averaging revealed small unitary excitatory postsynaptic currents in four of 39 cell pairs tested. The muscarine-induced inward current was attenuated by a phospholipase C (PLC) inhibitor, U73122, but not by a protein kinase C inhibitor, chelerythrine, or by the intracellular calcium chelator 1,2-bis(2-aminophenoxy) ethane-N,N,N',N'-tetra-acetic acid, suggesting that the current was associated with PLC in a protein kinase C- and Ca2+ -independent manner. The phosphatidylinositol 4-kinase inhibitor wortmannin (10 microm) reduced the recovery of the inward current, indicating that the recovery process was dependent on the removal of diacylglycerol and/or inositol 1,4,5 triphosphate or resynthesis of phospholipid phosphatidylinositol 4,5 bisphophate. Ratiometric measurement of intracellular calcium after cell loading with fura-2 demonstrated a muscarine-induced increase in calcium signal that originated mainly from intracellular stores. Thus, the cholinergic excitatory effect in striatal medium spiny neurones, which is important in motor disorders associated with altered cholinergic transmission in the striatum such as Parkinson's disease, is mediated through M1 receptors and the PLC-dependent pathway. PMID- 15341595 TI - An evolutionarily conserved mechanism for sensitization of soluble guanylyl cyclase reveals extensive nitric oxide-mediated upregulation of cyclic GMP in insect brain. AB - Soluble guanylyl cyclase (SGC) is the main receptor for the gaseous signalling molecule nitric oxide (NO) in vertebrates and invertebrates. Recently, a novel class of drugs that regulate mammalian SGC by NO-independent allosteric mechanisms has been identified [e.g. 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole, YC-1]. To assess the evolutionary conservation and hence the potential physiological relevance of these mechanisms, we have tested YC-1 on the brains of two model insects, the cockroach Periplaneta americana and the locust Schistocerca gregaria. YC-1 strongly potentiated the NO-induced elevation of total cyclic 3',5'-guanosine monophosphate (cGMP) and amplified the intensity and consistency of NO-induced cGMP-immunoreactivity in the brain. Our data indicate that the effect of YC-1 was independent of phosphodiesterase inhibition and thus mediated by direct sensitization of SGC. Immunohistopharmacology and co-labelling with antibodies against the SGC alpha-subunit confirmed that cGMP induced by co application of NO and YC-1 is predominantly attributable to SGC. The staggering number of NO-responsive neurons revealed by YC-1 suggests that previous studies may have considerably underestimated the number of cellular targets for NO in the insect brain. Moreover, a subset of these targets exhibited cGMP-immunoreactivity without application of exogenous NO, demonstrating that YC-1 can be exploited for visualization of physiological cGMP signals in response to endogenous NO production. In conclusion, our discovery that YC-1 is a potent sensitizer of insect SGC indicates that a NO-independent regulatory site is an evolutionarily conserved feature of SGC. Our findings add considerable momentum to the concept of an as yet unidentified endogenous ligand that regulates the gain of the NO cGMP signalling pathway. PMID- 15341596 TI - Activation of group I metabotropic glutamate receptors enhances efficacy of glutamatergic inputs to neonatal rat hypoglossal motoneurons in vitro. AB - Group I metabotropic glutamate receptors (mGluRs) are the main class of metabotropic receptors expressed in the hypoglossus nucleus. Their role in glutamatergic transmission was investigated using patch-clamp recording from motoneurons in a neonatal rat brainstem slice preparation. After pharmacological block of gamma-aminobutyric acid and glycine-mediated inhibition, under voltage clamp, the selective group I agonist (RS)-3,5-dihydroxyphenylglycine (DHPG) induced a motoneuron inward current by depressing a leak conductance, and strongly facilitated spontaneous glutamatergic synaptic currents. This effect was blocked by 7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOEt) and unaffected by 2-methyl-6-(phenylethynyl)pyridine hydrochloride (MPEP), indicating a role for subtype 1 mGluRs. The frequency but not the amplitude of miniature glutamatergic currents was also enhanced by DHPG. Currents elicited by puffer application of (RS)-alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) in the presence of tetrodotoxin were also unchanged, suggesting that DHPG facilitated release of glutamate. Glutamatergic currents evoked by electrically stimulating the dorsomedullary reticular column premotoneurons were, however, depressed by DHPG in a CPCCOEt-sensitive fashion. Neither CPCCOEt nor MPEP per se changed glutamatergic transmission. Under current clamp, even if DHPG depressed excitatory postsynaptic potentials, motoneuron spike threshold and time to peak were reduced so that facilitation of synaptic potential/spike coupling became apparent. We propose a wiring diagram to account for the differential action by DHPG on spontaneous and evoked transmission, based on the discrete distribution of subtype 1 mGluRs on glutamatergic afferents. Although under standard recording conditions there was insufficient ambient glutamate to activate mGluRs, such receptors were a powerful target to upregulate excitatory synaptic transmission and enhance signalling by hypoglossal motoneurons to tongue muscles. PMID- 15341597 TI - Chronic L-DOPA treatment increases extracellular glutamate levels and GLT1 expression in the basal ganglia in a rat model of Parkinson's disease. AB - There is growing experimental evidence for the implication of glutamate-mediated mechanisms both in the pathophysiology of Parkinson's disease and in the development of dyskinesias with long-term administration of L-3,4 dihydroxyphenylalanine (L-DOPA). However, the impact of this treatment on glutamate transmission in the basal ganglia has been poorly investigated. In this study, we examined the effects of 6-hydroxydopamine-induced lesion of nigral dopamine neurons with or without subsequent chronic L-DOPA treatment on several parameters of glutamate system function in the rat striatum and substantia nigra pars reticulata. All the lesioned animals treated with L-DOPA developed severe dyskinesias. Extracellular glutamate levels, measured by microdialysis in freely moving conditions, and gene expression of the glial glutamate transporter GLT1, assessed by in situ hybridization, were unaffected by dopamine lesion or L-DOPA treatment alone, but were both markedly increased on the lesion side of rats with subsequent L-DOPA treatment. No change in the expression of the vesicular glutamate transporters vGluT1 and vGluT2 was measured in striatum. These data show that chronic L-DOPA treatment leading to dyskinesias increases basal levels of glutamate function in basal ganglia. The L-DOPA-induced overexpression of GLT1 may represent a compensatory mechanism involving astrocytes to limit glutamate overactivity and subsequent toxic processes. PMID- 15341598 TI - Endogenous acetylcholine lowers the threshold for long-term potentiation induction in the CA1 area through muscarinic receptor activation: in vivo study. AB - Little is known how synaptically released endogenous ACh affects hippocampal synaptic plasticity in vivo. Here, we examined the role of cholinergic drive in the regulation of the induction of long-term potentiation (LTP) at basal dendrites in the CA1 area of the anaesthetized rat hippocampus. The non-subtype selective muscarinic acetylcholine receptor antagonist, scopolamine, (0.3 mg/kg, i.p.) inhibited the induction of LTP by weak, but not strong, high frequency conditioning stimulation. A relatively M1 subtype-selective receptor antagonist, pirenzepine, (50 nmol/5 microL, i.c.v.) also inhibited LTP induction by the weak protocol. As the medial septum (MS) is a major source of endogenous ACh in the hippocampus, we also examined the effect of high frequency pre-conditioning stimulation of the MS on LTP induction. The pre-conditioning MS tetanus reduced the threshold for LTP induction at basal synapses in a narrow time window. Such an effect of MS pre-conditioning was prevented by scopolamine, strong evidence of a direct MS control of LTP threshold through a mechanism dependent on muscarinic receptor activation. These results suggest that the cholinergic drive to the hippocampus is critically involved in the control of the LTP induction threshold in vivo. To the extent that LTP mechanisms may underlie certain types of learning and memory, the septo-hippocampal cholinergic regulation of synaptic plasticity may constitute an important target for the treatment of cognitive disorders associated with ACh deficits. PMID- 15341599 TI - Isoflurane modulates glutamatergic and GABAergic neurotransmission in the amygdala. AB - Attempts have been made to attribute the particular features of general anaesthesia such as hypnosis, analgesia, amnesia and autonomic stability to certain brain regions. In the present study, we examined the effects of the commonplace volatile anaesthetic isoflurane on synaptic transmission in an in vitro slice preparation of the murine amygdala. Despite the established role of this limbic structure in the formation of aversive memories, conditioned fear and anxiety, as well as pain processing and regulation of sympathetic tone, the influence of volatile anaesthetics on synaptic signalling has not yet been investigated in this region of the brain. Evoked postsynaptic currents were monitored from principal neurons in the basolateral nucleus of the amygdala by means of patch-clamp recording. The mixed postsynaptic currents were mediated by non-NMDA, NMDA, GABA A and GABA B receptors. Isoflurane added to the perfusion medium reduced the strength of synaptic signalling following the activation of non-NMDA, NMDA, and GABA B receptors, whereas the GABA A receptor-mediated responses were enhanced. The overall reduction of neuronal excitability was also reflected in a reduction of field potential amplitudes. Isoflurane neither changed the membrane resting potential nor the input resistance of principal neurons in the amygdala. The present results may contribute to the understanding of how stress reactions and long-lasting neuroplastic processes are suppressed under general anaesthesia. PMID- 15341600 TI - Functional mapping of GABA A receptor subtypes in the amygdala. AB - The physiological significance of the large diversity of GABA A receptors is poorly understood. Using mice, which carry a point mutation that renders specific subtypes of GABA A receptors diazepam insensitive, it was recently discovered that particular types of GABA A receptors are involved in specific, behaviorally relevant signaling pathways. We have used these mice to study inhibitory synaptic transmission in the amygdala. GABA A receptor-mediated inhibitory postsynaptic currents (IPSCs) per se were not affected by the point mutations. Their modulation by diazepam, however, was altered depending on the genotype of the mice studied. Based on the different responses to diazepam, we found that IPSCs in the lateral/basolateral amygdala were mediated by both alpha2- and alpha1 subunit-containing GABA A receptors whereas those in the central amygdala were mediated only by alpha2-subunit-containing GABA A receptors. Immunohistochemical staining corroborated these findings at a morphological level. To investigate a possible link between interneuron and receptor diversity, we selectively depressed release from the subset of GABAergic terminals carrying type 1 cannabinoid receptors. These receptors are known to modulate amygdala-mediated behavior. Application of a type 1 cannabinoid receptor agonist resulted in a selective reduction of inhibitory current mediated by alpha1-subunit-containing GABA A receptors. Mice with specific diazepam-insensitive GABA A receptor subtypes therefore provide a novel tool to investigate GABA A receptor distribution and the organization of inhibitory circuits at a functional level. The crucial role of the amygdala for the mediation of anxiety is in agreement with the part that alpha2-subunit-containing GABA A receptors play in anxiolysis and their important function in this area of the brain. PMID- 15341601 TI - Brain-derived neurotrophic factor controls functional differentiation and microcircuit formation of selectively isolated fast-spiking GABAergic interneurons. AB - GABAergic interneurons with high-frequency firing, fast-spiking (FS) cells, form synapses on perisomatic regions of principal cells in the neocortex and hippocampus to control the excitability of cortical networks. Brain-derived neurotrophic factor (BDNF) is essential for the differentiation of multiple interneuron subtypes and the formation of their synaptic contacts. Here, we examined whether BDNF, alone or in conjunction with sustained KCl-induced depolarization, drives functional FS cell differentiation and the formation of inhibitory microcircuits. Homogeneous FS cell cultures were established by target specific isolation using the voltage-gated potassium channel 3.1b subunit as the selection marker. Isolated FS cells expressed parvalbumin, were surrounded by perineuronal nets, formed immature inhibitory connections and generated slow action potentials at 12 days in vitro. Brain-derived neurotrophic factor (BDNF) promoted FS cell differentiation by increasing the somatic diameter, dendritic branching and the frequency of action potential firing. In addition, BDNF treatment led to a significant up-regulation of synaptophysin and vesicular GABA transporter expression, components of the synaptic machinery critical for GABA release, which was paralleled by an increase in synaptic strength. Long-term membrane depolarization alone was detrimental to dendritic branching. However, we observed that BDNF and KCl exerted additive effects, as reflected by the significantly accelerated maturation of synaptic contacts and high discharge frequencies, and was required for the formation of reciprocal connections between FS cells. Our results show that BDNF, along with membrane depolarization, is critical for FS cells to establish inhibitory circuitries during corticogenesis. PMID- 15341602 TI - GABAergic phenotypic differentiation of a subpopulation of subventricular derived migrating progenitors. AB - Olfactory bulb interneurons are continuously generated throughout development and in adulthood. These neurons are born in the subventricular zone (SVZ) and migrate along the rostral migratory stream into the olfactory bulb where they differentiate into local interneurons. To investigate the differentiation of GABAergic interneurons of the olfactory bulb we used a transgenic mouse which expresses green fluorescent protein (GFP) under the control of the glutamic acid decarboxylase 65 kDa (GAD65) promoter. During development and in adulthood GFP was expressed by cells in the SVZ and along the entire length of its rostral extension including the distal portion within the olfactory bulb. The occurrence of GAD65 mRNA in these zones was confirmed by PCR analysis on microdissected regions along the pathway. Polysialic acid neural cell adhesion molecule, a marker of migrating neuroblasts in adults, was coexpressed by the majority of the GFP-positive SVZ-derived progenitor cells. Cell tracer injections into the SVZ indicated that approximately 26% of migrating progenitor cells expressed GFP. These data show the early differentiation of migrating SVZ-derived progenitors into a GAD65-GFP-positive phenotype. These cells could represent a restricted lineage giving rise to GAD65-positive GABAergic olfactory bulb interneurons. PMID- 15341603 TI - GABAergic neurons participate in the brain's response to birdsong auditory stimulation. AB - Birdsong is a learned vocal behaviour that requires intact hearing for its development in juveniles and for its maintenance during adulthood. However, the functional organization of the brain circuits involved in the perceptual processing of song has remained obscure. Here we provide evidence that GABAergic mechanisms are an important component of these circuits and participate in the auditory processing of birdsong. We first cloned a zebra finch homologue of the gene encoding the 65-kDa isoform of glutamic acid decarboxylase (zGAD-65), a specific GABAergic marker, and conducted an expression analysis by in situ hybridization to identify GABAergic cells and to map their distribution throughout auditory telencephalic areas. The results showed that field L2, the caudomedial nidopallium (NCM) and the caudomedial mesopallium (CMM) contain a high number of GABAergic cells. Using patch-clamp brain slice recordings, we found abundant GABAergic mIPSCs in NCM. Pharmacological antagonism of mIPSCs induced large EPSC bursts, suggesting that tonic inhibition helps to stabilize NCM against runaway excitation via activation of GABA-A receptors. Next, using double fluorescence in situ hybridization and double immunocytochemical labelling, we demonstrated that large numbers of GABAergic cells in NCM and CMM show inducible expression of the transcriptional regulator ZENK in response to song auditory stimulation. These data provide direct evidence that GABAergic neurons in auditory brain regions are activated by song stimulation. Altogether, our results suggest that GABAergic mechanisms participate in auditory processing and perception, and might contribute to the memorization of birdsong. PMID- 15341604 TI - Contribution of GABAergic inhibition to synaptic responses and LTD early in postnatal development in the rat superior colliculus. AB - We studied the development of optic tract evoked field potentials (FP) in the rodent superior colliculus (SC) and the effect of GABA antagonists upon their development and upon induction of long-term depression (LTD). Brain slices were cut from Lister Hooded rats. The optic tract was stimulated while recording from the superficial grey layer. GABAergic inhibition was assessed by adding 100 microm picrotoxin and 3 microm CGP55845 antagonists to block GABA A,B,C receptors. LTD was induced with a 50 Hz, 20 s tetanus. At age P2, the FP consisted only of a presynaptic spike. The GABA antagonists had no effect. By P4, the FP consisted of a presynaptic spike, a longer latency population spike, and a field excitatory postsynaptic potential (fEPSP). The fEPSP was slightly prolonged by the GABA antagonists at this age. By P7-P14, a prominent FP with trailing fEPSP was recorded. The GABA antagonists usually had a large effect, with the fEPSP increasing in both amplitude and duration. A mature FP was usually recorded in P15-P23 slices where the GABA antagonist effect remained substantial. LTD could be induced in 17 of 30 control slices from rats aged P4-P26. The average fEPSP amplitude after tetanus was 77.9% of control. Pre-treatment with GABA antagonists produced a short-term potentiation (average 114.0%), rather than LTD, in 14 of 19 cases. This STP was followed by a more prolonged potentiation in 12 of the 14 cases. We conclude that GABAergic inhibitory circuits mature before eye opening and that GABA contributes to induction of LTD in the developing SC. PMID- 15341605 TI - Environmental enrichment in mice decreases anxiety, attenuates stress responses and enhances natural killer cell activity. AB - The importance of environment in the regulation of brain, behaviour and physiology has long been recognized in biological, social and medical sciences. Animals maintained under enriched conditions have clearly been shown to have better learning abilities than those maintained under standard conditions. However, the effects of environmental enrichment (EE) on immunity and emotionality have been less documented and remain questionable. Therefore, we investigated the effect of EE on natural killer (NK) cell activity, psychological stress responses and behavioural parameters. Male C3H mice were housed either in enriched or standard conditions for 6 weeks. Behaviour was then examined by the grip-strength test, staircase and elevated plus maze, and corticosterone levels and NK cell activity were measured. Furthermore, animals exposed to the stress paradigm, achieved by electric shock with reminders, were tested for freezing time in each reminder. Corticosterone levels were also measured. The EE mice showed decreased anxiety-like behaviour and higher activity compared to standard mice, as revealed by a greater percentage of time spent in the open arms of the elevated plus maze, and a higher rate of climbing the staircase. A shorter freezing time in the stress paradigm and no corticosterone level reactivity were measured in EE mice. In addition, NK cell activity in spleens of EE mice was higher than that demonstrated in those of standard mice. Thus, EE has a beneficial effect on anxiety-like behaviour, stress response and NK cell activity. The effect on NK cell activity is promising, due to the role of NK cells in host resistance. PMID- 15341606 TI - Corticostriatal brain-derived neurotrophic factor dysregulation in adult rats following prenatal stress. AB - Prenatal stress represents a well-established experimental protocol resembling some features of schizophrenia, including deficits in social interactions, disruption of prepulse inhibition and enhanced response to psychomotor stimulants. In order to evaluate molecular changes that could participate in long lasting effects on brain function, we analysed the effects of prenatal stress on the expression of brain-derived neurotrophic factor (BDNF), an important molecular determinant of synaptic plasticity and cellular homeostasis, in adult male rats under basal conditions as well as in response to a chronic stress. The main finding is that BDNF expression is reduced in the prefrontal cortex and striatum of prenatally stressed rats. Furthermore, when exposed to chronic stress in adulthood, these rats display an altered regulation of BDNF expression in these brain structures, implying that adverse life events during gestation may interfere with the expression and function of this neurotrophin at adulthood in a region-specific manner. The dysregulation of corticostriatal BDNF expression might thus contribute to permanent alterations in brain functions leading to heightened susceptibility to psychiatric disorders. PMID- 15341607 TI - Peripubertal environmental enrichment reverses the effects of maternal care on hippocampal development and glutamate receptor subunit expression. AB - Maternal care in the rat influences the development of cognitive function in the offspring through neural systems known to mediate activity-dependent synaptic plasticity. The offspring of mothers that exhibit increased levels of pup licking/grooming (high-LG mothers) show increased hippocampal N-methyl-D aspartate (NMDA) subunit mRNA expression, enhanced synaptogenesis and improved hippocampal-dependent spatial learning in comparison with animals reared by low LG mothers. The effects of reduced maternal care on cognitive function are reversed with peripubertal environmental enrichment; however, the neural mechanisms mediating this effect are not known. In these studies we exposed the offspring of high- and low-LG mothers to environmental enrichment from days 22 to 70 of life, and measured the expression of genes encoding for glutamate receptor subunits and synaptophysin expression as a measure of synaptic density. Environmental enrichment reversed the effects of maternal care on synaptic density and this effect was, in turn, associated with a reversal of the effect of maternal care on the NR2A and NR2B subunits of the NMDA receptor, as well as effects on (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor subunits. Finally, direct infusion of an NR2B-specific NMDA receptor antagonist into the hippocampus eliminated the effects of maternal care on spatial learning/memory in the Morris water maze. These findings suggest that: (1) the effects of maternal care are mediated by changes in NR2B gene expression; and (2) that environmental enrichment reverses the effects of reduced maternal care through the same genomic target, the NR2B gene, and possibly effects on other subunits of the NMDA and AMPA receptors. PMID- 15341608 TI - Regional pattern of metabolic activation is reflected in the sleep EEG after sleep deprivation combined with unilateral whisker stimulation in mice. AB - Regional differences in EEG slow wave activity (SWA) during sleep after sleep deprivation (SD) may be a consequence of differential metabolic activation of cortical areas. We investigated the relationship between the regional EEG dynamics and 2-deoxyglucose (DG) uptake after SD in mice. Six hours' SD were combined with natural unilateral whisker stimulation in an enriched environment to selectively activate the barrel cortex and motor areas. As expected, an interhemispheric asymmetry of 2-DG uptake was found in the barrel cortex immediately after SD. To test whether sleep contributes to recovery of the asymmetry, the stimulation was followed by either undisturbed sleep or by an additional SD. The asymmetry vanished after recovery sleep but also after the additional period of wakefulness without stimulation. In addition, relative 2-DG uptake in the primary motor cortex and retrosplenial area was significantly higher immediately after the SD than after the additional sleep or wakefulness, whereas no other region differed between the groups. Whisker stimulation elicited a greater increase in EEG SWA during non rapid eye movement sleep in the stimulated hemisphere than in the control hemisphere; this increase lasted for 10 h. Within a hemisphere, the initial increase in SWA was higher in the frontal than in the parietal derivation. We conclude that the regional SWA differences during sleep are use-dependent and may be related to the regional pattern of metabolism during the previous waking episode. However, the regional metabolic recovery is not dependent on sleep, and is not directly reflected in changes in SWA during sleep. PMID- 15341609 TI - Attention and executive function deficits in chronic low-dose MPTP-treated non human primates. AB - Parkinson's disease (PD) is a complex disorder consisting of motor deficits coupled with dysfunction in cognitive domains that are dependent upon the integrity of the frontal lobes and/or the fronto-striatal axis. Although it is increasingly acknowledged that PD patients have attentional and executive function deficits, it has been difficult to model these in nonhuman primates because of the nature of the cognitive tasks that have been used previously. The present studies were conducted to further define the nature of the cognitive impairment in a nonhuman primate model of early parkinsonism consequent to chronic low dose MPTP exposure and to further validate this model in monkeys trained to perform a battery of attentional and executive function tasks. Following chronic low dose MPTP exposure, monkeys developed deficits in maintenance of a response set as well problems in shifting attentional sets, suggesting decreased mental flexibility. On other tasks inattentiveness, an impaired ability to sustain spatial attention or to focus attention, a deficit in motor readiness and planning, and impaired time estimation were also observed. These results provide direct evidence of attention and executive function deficits in a nonhuman primate model of early parkinsonism. Based on these findings, we suggest that in addition to being useful for studying the cognitive deficits related to early PD and for developing new therapeutics for these problems, this model and these testing procedures may also provide a useful large animal model for studying attention deficit disorder and for developing new therapeutics for that condition as well. PMID- 15341610 TI - Independent coding of connected environments by place cells. AB - Place cells are hippocampal neurons that have a strong location-specific firing activity in the rat's current environment. Collectively, place cells also provide a signature of the rat's environment as their ensemble activity is markedly different when recorded in distinct apparatuses. This phenomenon, referred to as 'remapping', suggests that each environment activates a different hippocampal map. In this study, we sought to determine the independence of such maps. In Experiment 1, we used a cylinder apparatus that was divided into two equal halves by a central barrier with an aperture allowing the rat to freely commute between the two sides. A local change in one side failed to induce field remapping in the changed side, thus precluding any significant conclusion to be drawn. We therefore designed Experiment 2 in which place cells were first recorded while rats explored three distinct high-walled boxes. Most cells had distinctive firing fields in each box. A runway was then added to connect two initially unrelated boxes. This manipulation altered the firing of some cells but the fields in each box were still clearly distinguishable. The final manipulation consisted of changing one box and allowing the rat to commute freely between the changed and unchanged boxes. While the firing fields remapped in the changed box, they were most usually unaltered in the unchanged box. These results suggest that the hippocampus holds a set of independent maps for each box, and that each specific map is activated mainly according to the rat's current sensory environment. PMID- 15341611 TI - Directed interactions between visual areas and their role in processing image structure and expectancy. AB - During sensory processing, cortical areas continuously exchange information in different directions along the hierarchy. The functional role of such interactions, however, has been the subject of various proposals. Here, we investigate the role of bottom-up and top-down interactions in processing stimulus structure and their relation to expected events. Applying multivariate autoregressive methods to local field potentials recorded in alert cats, we quantify directed interactions between primary (A17/18) and higher (A21) visual areas. A trial-by-trial analysis yields the following findings. To assess the role of interareal interactions in processing stimulus structure, we recorded in naive animals during stimulation with natural movies and pink noise stimuli. The overall interactions decrease compared with baseline for both stimuli. To investigate whether forthcoming events modulate interactions, we recorded in trained animals viewing two stimuli, one of which had been associated with a reward. Several results support such modulations. First, the interactions increase compared with baseline and this increase is not observed in a context where food was not delivered. Second, these stimuli have a differential effect on top-down and bottom-up components. This difference is emphasized during the stimulus presentation and is maximal shortly before the possible reward. Furthermore, a spectral decomposition of the interactions shows that this asymmetry is most dominant in the gamma frequency range. Concluding, these results support the notion that interareal interactions are more related to an expectancy state rather than to processing of stimulus structure. PMID- 15341612 TI - The frontal predominance in human EEG delta activity after sleep loss decreases with age. AB - Sleep loss has marked and selective effects on brain wave activity during subsequent recovery sleep. The electroencephalogram (EEG) responds to sleep deprivation with a relative increase in power density in the delta and theta range during non-rapid eye movement sleep. We investigated age-related changes of the EEG response to sleep deprivation along the antero-posterior axis (Fz, Cz, Pz, Oz) under constant routine conditions. Both healthy young (20-31 years) and older (57-74 years) participants manifested a significant relative increase in EEG power density in the delta and theta range after 40 h of sleep deprivation, indicating a sustained capacity of the sleep homeostat to respond to sleep loss in ageing. However, the increase in relative EEG delta activity (1.25-3.75 Hz) following sleep deprivation was significantly more pronounced in frontal than parietal brain regions in the young, whereas such a frontal predominance was diminished in the older volunteers. This age-related decrease of frontal delta predominance was most distinct at the beginning of the recovery sleep episode. Furthermore, the dissipation of homeostatic sleep pressure during the recovery night, as indexed by EEG delta activity, exhibited a significantly shallower decline in the older group. Activation of sleep regulatory processes in frontal brain areas by an extension of wakefulness from 16 to 40 h appears to be age dependent. These findings provide quantitative evidence for the hypothesis that frontal brain regions are particularly vulnerable to the effects of elevated sleep pressure ('prefrontal tiredness') and ageing ('frontal ageing'). PMID- 15341613 TI - Neural contributions to the motivational control of appetite in humans. AB - The motivation to eat in humans is a complex process influenced by intrinsic mechanisms relating to the hunger and satiety cascade, and extrinsic mechanisms based on the appetitive incentive value of individual foods, which can themselves induce desire. This study was designed to investigate the neural basis of these two factors contributing to the control of motivation to eat within the same experimental design using positron emission tomography. Using a novel counterbalanced approach, participants were scanned in two separate sessions, once after fasting and once after food intake, in which they imagined themselves in a restaurant and considered a number of items on a menu, and were asked to choose their most preferred. All items were tailored to each individual and varied in their incentive value. No actual foods were presented. In response to a hungry state, increased activation was shown in the hypothalamus, amygdala and insula cortex as predicted, as well as the medulla, striatum and anterior cingulate cortex. Satiety, in contrast, was associated with increased activation in the lateral orbitofrontal and temporal cortex. Only activity in the vicinity of the amygdala and orbitofrontal cortex was observed in response to the processing of extrinsic appetitive incentive information. These results suggest that the contributions of intrinsic homeostatic influences, and extrinsic incentive factors to the motivation to eat, are somewhat dissociable neurally, with areas of convergence in the amygdala and orbitofrontal cortex. The findings of this study have implications for research into the underlying mechanisms of eating disorders. PMID- 15341614 TI - RFa-related peptides are algogenic: evidence in vitro and in vivo. AB - RFamide (RFa)-related peptides modulate pain processing in the mammalian CNS. The effects of these peptides are generally considered as 'anti-opioid'. They also decrease the rate of desensitization of acid-sensing ionic channels (ASICs), putative nociceptors in dorsal root ganglia neurons [C. Askwith et al. (2000) Neuron, 26, 133-141]. We have tested the role of mollusc-derived peptide, FMRFa (Phe-Met-Arg-Phe-amide) and its synthetic analogues in peripheral nociception. Here we demonstrate that RFa-related peptides powerfully excite the majority of C fibres in the skin-nerve preparation of rat: 76% of 55 tested fibres with the conduction velocity below 2 m/s responded with long-lasting discharges to the application of peptides (20 microm). When injected subcutaneously in vivo (mice), they initiate nociceptive behaviour. We confirm the data on humans [S. Ugawa et al. (2002) J. Clin. Invest., 110, 1185-1190]: the activation of C-fibres by acid is inhibited by channel blocker of ASICs, amiloride. However, there is no correlation in the sensitivity of C-fibres to RFa peptides, protons and amiloride: 74% of tested RFa-sensitive C-fibres were insensitive to protons and in 67% of cases the response to peptides was insensitive to amiloride. Thus, powerful excitatory/algogenic action of RFa-related peptides cannot be interpreted solely in terms of their interaction with ASICs. The peptides do not activate any conductance in the somatic membrane of dorsal root ganglion neurons of rats and probably affect still unidentified molecular target(s) responsible for nociceptive signalling. PMID- 15341615 TI - Effective use of fluorides for the prevention of dental caries in the 21st century: the WHO approach. AB - Despite great improvements in the oral health of populations across the world, problems still persist particularly among poor and disadvantaged groups in both developed and developing countries. According to the World Oral Health Report 2003, dental caries remains a major public health problem in most industrialized countries, affecting 60-90% of schoolchildren and the vast majority of adults. Although it appears that dental caries is less common and less severe in developing countries of Africa, it is anticipated that the incidence of caries will increase in several countries of that continent, due to changing living conditions and dietary habits, and inadequate exposure to fluorides. Research on the oral health effects of fluoride started around 100 years ago; the focus has been on the link between water and fluorides and dental caries and fluorosis, topical fluoride applications, fluoride toothpastes, and salt and milk fluoridation. Most recently, efforts have been made to summarize the extensive database through systematic reviews. Such reviews concluded that water fluoridation and use of fluoride toothpastes and mouthrinses significantly reduce the prevalence of dental caries. WHO recommends for public health that every effort must be made to develop affordable fluoridated toothpastes for use in developing countries. Water fluoridation, where technically feasible and culturally acceptable, has substantial advantages in public health; alternatively, fluoridation of salt and milk fluoridation schemes may be considered for prevention of dental caries. PMID- 15341616 TI - Caries management for institutionalized elders using fluoride and chlorhexidine mouthrinses. AB - OBJECTIVES: The effectiveness of either a 0.2% neutral sodium fluoride (NaF) solution or a 0.12% chlorhexidine (CHX) solution as a daily mouthrinse for controlling caries was tested against a placebo rinse in this 2-year randomized clinical trial among elders in long-term care (LTC) facilities. METHODS: At baseline, 369 recruits were examined clinically for caries and allocated randomly to one of the mouthrinse groups. RESULTS: After 2 years, 116 participants remained in the trial. The prevalence of caries and the dental status of the groups were similar at baseline and after 2 years. On average, each group lost less than one tooth per person, but the fluoride group compared with the others had significantly less caries and significantly more reversals from carious to sound dental surfaces at the end of the trial. CONCLUSIONS: We conclude that 0.2% neutral NaF mouthrinse every day does reduce the incidence of caries among elders in LTC facilities. PMID- 15341617 TI - Race/ethnicity and untreated dental caries: the impact of material and behavioral factors. AB - OBJECTIVE: To use nationally representative data, group variables into categories of material and behavioral factors, and assess their relative contribution to racial/ethnic variation in untreated caries. METHODS: Participants were from the Third National Health and Nutrition Examination Survey (NHANES III), aged 20-50 years. Material factors were income, education, employment status, dental insurance status, and urban residence. Behavioral factors were marital status, tobacco use, alcohol use, obesity, and social support. All models were additionally adjusted for age, gender, and quartile of missing teeth. The outcome was three or more carious teeth. RESULTS: Non-Hispanic blacks and Mexican Americans displayed excess risk of untreated caries compared with non-Hispanic whites when adjusted for age, sex, and missing teeth (adjusted odds ratios 1.73 and 1.69, respectively). The addition of behavioral factors to this model resulted in virtually no changes in the adjusted odds ratios for race/ethnicity and untreated caries. When material factors were added to the basic model the excess risk for untreated caries among non-Hispanic blacks was reduced by approximately 21% and that of Mexican-Americans was no longer statistically significant compared with non-Hispanic whites (adjusted odds ratios 1.36 and 0.83, respectively). CONCLUSIONS: Much of the excess risk for untreated dental caries among non-Hispanic blacks and Mexican-Americans compared with non-Hispanic whites was eliminated when material factors were controlled, while no risk reductions were observed when behavioral factors were controlled. Addressing material factors may provide greater reductions in untreated caries disparities than behavioral interventions, and these risk reductions may vary with racial/ethnic group. PMID- 15341618 TI - The relationship between dental caries and dental fluorosis in areas with moderate- and high-fluoride drinking water in Ethiopia. AB - OBJECTIVE: The aim of the study is to assess the relationship between caries and dental fluorosis in Ethiopian children living in Rift Valley areas known for endemic fluorosis. METHOD: A total of 306 children (12-15 years old), selected from areas with moderate (0.3-2.2 mg/l), or high (10-14 mg/l) fluoride concentration in the drinking water were interviewed and examined for caries and dental fluorosis. Scorings were recorded according to the DMF system, and the Thylstrup-Fejerskov (TF) Index. RESULTS: Prevalence of dental fluorosis (TF-score > or = 1) was 91.8% (moderate area) and 100% (high-fluoride area). The corresponding caries prevalence and mean DMFT in the areas were 45.3% versus 61.6%, and 1.2 versus 1.8, respectively. Age and severity of dental fluorosis were found to be independent predictors for DMFT > or = 1. When compared with 12 year olds with TF-scores 0-4, odds ratios were 3.0 (95% CI 1.6-5.7) and 2.0 (95% CI 1.2-3.2) if TF-scores were > or = 5 and age 13-15 years, respectively. A positive relationship between caries and fluorosis was observed across tooth types in both areas. The percentage of children with DMFT > or = 1 was highest in groups with TF-score > or = 5 in the second molar, followed by the first molar. CONCLUSION: The present findings indicate that the second molar is the tooth most severely affected by dental fluorosis and dental caries. Dental caries increased with increasing severity of dental fluorosis, both in moderate- and high-fluoride areas. Thus, a positive relationship between dental caries and dental fluorosis was observed across various tooth types, in both areas. PMID- 15341619 TI - Socioeconomic inequalities in oral health in childhood and adulthood in a birth cohort. AB - OBJECTIVES: To determine whether adult oral health is predicted by (a) childhood socioeconomic advantage or disadvantage (controlling for childhood oral health), or (b) oral health in childhood (controlling for childhood socioeconomic advantage or disadvantage), and whether oral health in adulthood is affected by changes in socioeconomic status (SES). METHODS: Participants in a longstanding cohort study underwent systematic dental examination for dental caries and tooth loss at ages 5 and 26 years. The examination at age 26 years included the collection of data on periodontal attachment loss and plaque level. Childhood SES was determined using parental occupation, and adult SES was determined from each study member's occupation at age 26 years. Regression models were used to test the study hypotheses. RESULTS: Complete data were available for 789 individuals (47.4% female). After controlling for childhood oral health, those who were of low SES at age 5 years had substantially greater mean DFS and DS scores by age 26 years, were more likely to have lost a tooth in adulthood because of caries, and had greater prevalence and extent of periodontitis. A largely similar pattern was observed (after controlling for childhood SES) among those with greater caries experience at age 5 years. For almost all oral health indicators examined, a clear gradient was observed of greater disease at age 26 years across socioeconomic trajectory groups, in the following order of ascending disease severity and prevalence: 'high-high', 'low-high' (upwardly mobile), 'high-low' (downwardly mobile) and 'low-low'. CONCLUSION: Adult oral health is predicted by not only childhood socioeconomic advantage or disadvantage, but also by oral health in childhood. Changes in socioeconomic advantage or disadvantage are associated with differing levels of oral health in adulthood. The life-course approach appears to be a useful paradigm for understanding oral health disparities. PMID- 15341620 TI - Stability of oral health-related behaviour in a Norwegian cohort between the ages of 15 and 23 years. AB - OBJECTIVE: To assess the stability in self-reported oral health behaviour in a Norwegian cohort between the ages of 15 and 23 years. METHODS: Self-administered questionnaires were used as part of a longitudinal cohort study. In 1992, a representative sample of 963 15-year-old adolescents participated, of which 676 (70%) and 567 (58%) remained in the study at ages 18 and 23 years. A total of 389 (40% of baseline) participated at each data collection, i.e. at ages 15, 18, 19, 21 and 23 years. RESULTS: General linear model (GLM) repeated-measures anova revealed statistically significant main effect of time with respect to soft drink and sweet consumption (F = 22.4, P < 0.001 and F = 4.3, P < 0.05, respectively). Adjusted mean scale scores of soft drink intake increased from 2.3 at age 15 years to 3.4 at age 23 years. The corresponding figures for consumption of sweets were 2.6 and 2.8. Two-way interactions achieved statistical significance with gender for soft drink consumption and toothbrushing. GLM repeated-measures with each gender revealed that soft drink consumption increased with time more extensively in boys (from 2.9 to 4.2, F = 13.5, P < 0.001) than in girls (from 1.9 to 2.6, F = 8.1, P < 0.001). Tracking or maintenance across time of the relative ranking at age 15 years occurred with all the four behaviours investigated. A total of 68-92% remained active and inactive regarding soft drink and sweet consumption, flossing and toothbrushing. CONCLUSION: The results provide evidence of tracking and early consolidation of oral health behaviour. This adds support for the assumption given for early intervention to prevent oral diseases. PMID- 15341621 TI - Perception of general and oral health in White and African American adults: assessing the effect of neighborhood socioeconomic conditions. AB - OBJECTIVES: This study investigates the independent and joint effects of family income and neighborhood socioeconomic status (SES) on general health and oral health before and after controlling for traditional risk factors in a representative sample of adults aged 18+ years residing in the Detroit tri-county area, Michigan. METHODS: Individuals data were obtained through interviews, while neighborhood data came from the 1990 US Census. SUDAAN was used to accommodate the complex sampling design and correlation of outcomes within the same neighborhoods. RESULTS: Whites in disadvantaged neighborhoods were four times more likely to rate their oral health as fair or poor [odds ratio (OR): 4.0; 95% confidence intervals (CI): 1.6-10.3] than their counterparts in advantaged neighborhoods. When evaluating the joint effects of family income and neighborhood SES, low-income Whites in disadvantaged neighborhoods were six times more likely to rate their oral health as fair or poor (OR: 6.4; 95% CI: 1.6-26.5) than their high-income counterparts in advantaged neighborhoods. The odds of rating general health as fair or poor was six times greater in low-income African Americans in disadvantaged neighborhoods (OR: 6.1; 95% CI: 1.6-23.8) than high income African Americans in advantaged neighborhoods. Similarly, low-income African Americans in disadvantaged neighborhoods were almost three times (OR: 2.8; 95% CI: 1.0-8.1) more likely to rate their oral health as fair/poor than high-income African Americans in advantaged neighborhoods. CONCLUSIONS: SES conditions at the neighborhood-level, independently or jointly with individual level income, appear to be important in evaluating racial/ethnic differences in self-rated oral health. Neighborhood conditions could tap into constructs not captured by individual-level variables on self-rated oral health. PMID- 15341622 TI - Changes in dental status over 10 years in 80-year-old people: a prospective cohort study. AB - OBJECTIVES: The aim of the present study was to follow-up the condition of the teeth over a sufficiently long period. METHODS: Dental examinations were a part of a multidisciplinary 10-year cohort study on the elderly. These examinations were made in 1990 (n = 226), 1995 (n = 90) and 2000 (n = 65) for the entire population born in 1910 and living in Jyvaskyla, Finland. The subjects were divided into two categories, dentate (one tooth or more) and edentulous. RESULTS: The results showed that men had more intact teeth and lower DMF scores than women, but the differences diminished during the follow-up period. The number of remaining and filled teeth of those women who took part in all three phases of the present study was higher than that of those who died during the follow-up. In men the DMF scores showed the opposite trend. The most significant deterioration during the 10-year follow-up was found in the number of teeth and DMF scores in men and in the number of remaining and filled teeth in women. CONCLUSION: Among men, in particular, significant changes in oral health status could be seen even between 80 and 90 years of age. Hence, regardless of advanced age, a subject should be motivated by the oral health care team to seek regular dental treatment. PMID- 15341623 TI - Preventing tobacco use in Norwegian dental practice. AB - OBJECTIVES: To determine dentists' and dental hygienists' intervention activity towards patients who smoke or use snus (oral moist snuff), and to establish which factors impede interventions and cause variations in approach. METHODS: A questionnaire was mailed to a sample of 1500 dentists (response rate: 68%) and all dental hygienists in the country (522 in all; response rate: 61%). RESULTS: Dental hygienists conversed with patients on smoking habits on average 18 min/week, while the dentists spent 13 min doing the same. The issue of snus-use was discussed, on average, for 3 min. In eight of 10 consultations with patients suffering from tobacco-induced disorders in the oral cavity, the dentists/dental hygienists raised the subject of smoking habits with the patient. In cases without visible tobacco-induced symptoms, inquiries were made concerning smoking habits in three of 10 dentist consultations and four of 10 consultations with dental hygienists. For first-time consultations, six of 10 were queried concerning their smoking habits by their dentist, while dental hygienists enquired in seven of 10 cases. Self-reported skills, perceived barriers and attitudes explained far more of the variance in intervention impact than background variables. There were moderate differences between dentists and dental hygienists. CONCLUSIONS: There is room for improvement in smoking and snus-use prevention efforts in the dental sector. If staff is to be rid of their misconceptions regarding the efficacy of intervention, it is important to inform them about the encouraging results at the population level. PMID- 15341625 TI - DOMINO1, a member of a small plant-specific gene family, encodes a protein essential for nuclear and nucleolar functions. AB - Arabidopsis embryos carrying the domino1 mutation grow slowly in comparison with wild type embryos and as a consequence reach only the globular stage at desiccation. The primary defect of the mutation at the cellular level is the large size of the nucleolus that can be observed soon after fertilization in the nuclei of both the embryo and the endosperm. The ultrastructure of mutant nucleoli is drastically different from wild type and points to a fault in ribosome biogenesis. DOMINO1 encodes a protein, which belongs to a plant-specific gene family sharing a common motif of unknown function, present in the tomato DEFECTIVE CHLOROPLASTS AND LEAVES (LeDCL) protein. Using a GFP protein fusion, we show that DOMINO1 is targeted to the nucleus. We propose that inactivation of DOMINO1 has a negative effect on ribosome biogenesis and on the rate of cell division. PMID- 15341626 TI - Transport of ricin and 2S albumin precursors to the storage vacuoles of Ricinus communis endosperm involves the Golgi and VSR-like receptors. AB - We have studied the transport of proricin and pro2S albumin to the protein storage vacuoles of developing castor bean (Ricinus communis L.) endosperm. Immunoelectron microscopy and cell fractionation reveal that both proteins travel through the Golgi apparatus and co-localize throughout their route to the storage vacuole. En route to the PSV, the proteins co-localize in large (>200 nm) vesicles, which are likely to represent developing storage vacuoles. We further show that the sequence-specific vacuolar sorting signals of both proricin and pro2SA bind in vitro to proteins that have high sequence similarity to members of the VSR/AtELP/BP-80 vacuolar sorting receptor family, generally associated with clathrin-mediated traffic to the lytic vacuole. The implications of these findings in relation to the current model for protein sorting to storage vacuoles are discussed. PMID- 15341627 TI - Characterization of AtCHX17, a member of the cation/H+ exchangers, CHX family, from Arabidopsis thaliana suggests a role in K+ homeostasis. AB - The Arabidopsis genome contains many sequences annotated as encoding H(+)-coupled cotransporters. Among those are the members of the cation:proton antiporter-2 (CPA2) family (or CHX family), predicted to encode Na(+),K(+)/H(+) antiporters. AtCHX17, a member of the CPA2 family, was selected for expression studies, and phenotypic analysis of knockout mutants was performed. AtCHX17 expression was only detected in roots. The gene was strongly induced by salt stress, potassium starvation, abscisic acid (ABA) and external acidic pH. Using the beta glucuronidase reporter gene strategy and in situ RT-PCR experiments, we have found that AtCHX17 was expressed preferentially in epidermal and cortical cells of the mature root zones. Knockout mutants accumulated less K(+) in roots in response to salt stress and potassium starvation compared with the wild type. These data support the hypothesis that AtCHX17 is involved in K(+) acquisition and homeostasis. PMID- 15341628 TI - Adjustment of diurnal starch turnover to short days: depletion of sugar during the night leads to a temporary inhibition of carbohydrate utilization, accumulation of sugars and post-translational activation of ADP-glucose pyrophosphorylase in the following light period. AB - A larger proportion of the fixed carbon is retained as starch in the leaf in short days, providing a larger store to support metabolism and carbon export during the long night. The mechanisms that facilitate this adjustment of the sink source balance are unknown. Starchless pgm mutants were analysed to discover responses that are triggered when diurnal starch turnover is disturbed. Sugars accumulated to high levels during the day, and fell to very low levels by the middle of the night. Sugars rose rapidly in the roots and rosette after illumination, and decreased later in the light period. Global transcript profiling revealed only small differences between pgm and Col0 at the end of the day but large differences at the end of the night, when pgm resembled Col0 after a 4-6 h prolongation of the night and many genes required for biosynthesis and growth were repressed [Plant J. 37 (2004) 914]. It is concluded that transient sugar depletion at the end of the night inhibits carbon utilization at the start of the ensuing light period. A second set of experiments investigated the stimulation of starch synthesis in response to short days in wild-type Col0. In short days, sugars were very low in the roots and rosette at the end of the dark period, and after illumination accumulated rapidly in both organs to levels that were higher than in long days. The response resembles pgm, except that carbohydrate accumulated in the leaf as starch instead of sugars. A similar response was found after transfer from long to short days. Inclusion of sugar in the rooting medium attenuated the stimulation of starch synthesis. Post translational activation of ADP-glucose pyrophosphorylase (AGPase) was increased in pgm, and in Col0 in short days. It is concluded that starch synthesis is stimulated in short day conditions because sugar depletion at the end of the night triggers a temporary inhibition of growth and carbohydrate utilization in the first part of the light period, leading to transient accumulation of sugar and activation of AGPase. PMID- 15341629 TI - A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway. AB - Arabidopsis thaliana RD26 cDNA, isolated from dehydrated plants, encodes a NAC protein. Expression of the RD26 gene was induced not only by drought but also by abscisic acid (ABA) and high salinity. The RD26 protein is localized in the nucleus and its C terminal has transcriptional activity. Transgenic plants overexpressing RD26 were highly sensitive to ABA, while RD26-repressed plants were insensitive. The results of microarray analysis showed that ABA- and stress inducible genes are upregulated in the RD26-overexpressed plants and repressed in the RD26-repressed plants. Furthermore, RD26 activated a promoter of its target gene in Arabidopsis protoplasts. These results indicate that RD26 functions as a transcriptional activator in ABA-inducible gene expression under abiotic stress in plants. PMID- 15341630 TI - Intron retention is a major phenomenon in alternative splicing in Arabidopsis. AB - Alternative splicing (AS) combines different transcript splice junctions that result in transcripts with shuffled exons, alternative 5' or 3' splicing sites, retained introns and different transcript termini. In this way, multiple mRNA species and proteins can be created from a single gene expanding the potential informational content of eukaryotic genomes. Search algorithms of AS forms in a variety of Arabidopsis databases showed they contained an unusually high fraction of retained introns (above 30%), compared with 10% that was reported for humans. The preponderance of retained introns (65%) were either part of open reading frames, present in the UTR region or present as the last intron in the transcript, indicating that their occurrence would not participate in non-sense mediated decay. Interestingly, the functional distribution of the transcripts with retained introns is skewed towards stress and external/internal stimuli related functions. A sampling of the alternative transcripts with retained introns were confirmed by RT-PCR and were shown to co-purify with polyribosomes, indicating their nuclear export. Thus, retained introns are a prominent feature of AS in Arabidopsis and as such may play a regulatory function. PMID- 15341631 TI - Biological functions of ent- and syn-copalyl diphosphate synthases in rice: key enzymes for the branch point of gibberellin and phytoalexin biosynthesis. AB - Rice (Oryza sativa L.) produces ent-copalyl diphosphate (ent-CDP) and syn-CDP as precursors for several classes of phytoalexins and the phytohormones, gibberellins (GAs). It has recently been shown that a loss-of-function mutation of OsCPS1, a gene encoding a putative ent-CDP synthase, results in a severely GA deficient dwarf phenotype in rice. To clarify the biological functions of the ent and syn-CDP synthases involved in the biosynthesis of phytoalexins and/or GAs, we isolated two cDNAs, OsCyc1 and OsCyc2, encoding putative diterpene cyclases from ultraviolet (UV)-irradiated rice leaves (cv. Nipponbare). The production of phytoalexins in rice leaves is known to be highly induced by UV treatment. Using a bacterial expression system, we demonstrated that OsCyc1 encodes syn-CDP synthase and that OsCyc2 and OsCPS1 encode ent-CDP synthase. The level of expression of the OsCyc1 and OsCyc2 transcripts in rice leaves increased drastically in response to UV treatment, whereas expression of the OsCPS1 transcript was not induced by UV light. These results suggest that OsCyc1, OsCyc2 and OsCPS1 are responsible for the biosynthesis of momilactones A and B and oryzalexin S, oryzalexins A-F and phytocassanes A-E, and GAs, respectively. Our results strongly suggest the presence of two ent-CDP synthase isoforms in rice, one that participates in the biosynthesis of GAs and a second that is involved in the biosynthesis of phytoalexins. PMID- 15341632 TI - Purification and cloning of an esterase from the weed black-grass (Alopecurus myosuroides), which bioactivates aryloxyphenoxypropionate herbicides. AB - Carboxyesterases which activate aryloxyphenoxypropionate (AOPP) graminicides to their bioactive herbicidal acids by hydrolysing the respective ester precursors have been identified in black-grass (Alopecurus myosuroides), a problem weed of cereal crops in Northern Europe. The dominant 40 kDa carboxyesterase was purified 1700-fold and identified as a serine hydrolase by affinity labelling with a biotinylated fluorophosphonate suicide substrate. MS-MS sequencing of a peptide digest identified it to be a member of the GDSL family of serine hydrolases. The full-length A. myosuroides hydrolase (Amgdsh1) was cloned by RACE-PCR and expressed in the yeast Pichia pastoris as a secreted enzyme. Expression was associated with activity towards AOPP esters. AmGDSH1 was predicted to be glycosylated and exported to the apoplast in planta. Based on the analysis of related sequences in monocotyledonous plants an alternative classification of the GDSL plant hydrolase superfamily is suggested and their importance in endogenous metabolism and herbicide bioactivation in crops and weeds discussed. PMID- 15341633 TI - Freezing-sensitive tomato has a functional CBF cold response pathway, but a CBF regulon that differs from that of freezing-tolerant Arabidopsis. AB - Many plants increase in freezing tolerance in response to low temperature, a process known as cold acclimation. In Arabidopsis, cold acclimation involves action of the CBF cold response pathway. Key components of the pathway include rapid cold-induced expression of three homologous genes encoding transcriptional activators, CBF1, 2 and 3 (also known as DREB1b, c and a, respectively), followed by expression of CBF-targeted genes, the CBF regulon, that increase freezing tolerance. Unlike Arabidopsis, tomato cannot cold acclimate raising the question of whether it has a functional CBF cold response pathway. Here we show that tomato, like Arabidopsis, encodes three CBF homologs, LeCBF1-3 (Lycopersicon esculentum CBF1-3), that are present in tandem array in the genome. Only the tomato LeCBF1 gene, however, was found to be cold-inducible. As is the case for Arabidopsis CBF1-3, transcripts for LeCBF1-3 did accumulate in response to mechanical agitation, but not in response to drought, ABA or high salinity. Constitutive overexpression of LeCBF1 in transgenic Arabidopsis plants induced expression of CBF-targeted genes and increased freezing tolerance indicating that LeCBF1 encodes a functional homolog of the Arabidopsis CBF1-3 proteins. However, constitutive overexpression of either LeCBF1 or AtCBF3 in transgenic tomato plants did not increase freezing tolerance. Gene expression studies, including the use of a cDNA microarray representing approximately 8000 tomato genes, identified only four genes that were induced 2.5-fold or more in the LeCBF1 or AtCBF3 overexpressing plants, three of which were putative members of the tomato CBF regulon as they were also upregulated in response to low temperature. Additional experiments indicated that of eight tomato genes that were likely orthologs of Arabidopsis CBF regulon genes, none were responsive to CBF overexpression in tomato. From these results, we conclude that tomato has a complete CBF cold response pathway, but that the tomato CBF regulon differs from that of Arabidopsis and appears to be considerably smaller and less diverse in function. PMID- 15341634 TI - High humidity suppresses ssi4-mediated cell death and disease resistance upstream of MAP kinase activation, H2O2 production and defense gene expression. AB - The Arabidopsis ssi4 mutant, which exhibits spontaneous lesion formation, constitutive expression of pathogenesis-related (PR) genes and enhanced resistance to virulent bacterial and oomycete pathogens, contains a gain-of function mutation in a TIR-NBS-LRR type R gene. Epistatic analyses revealed that both PR gene expression and disease resistance are activated via a salicylic acid (SA)- and EDS1-dependent, but NPR1- and NDR1-independent signaling pathway. In this study, we demonstrate that in moderate relative humidity (RH; 60%), the ssi4 mutant accumulates H(2)O(2) and SA prior to lesion formation and displays constitutive activation of the MAP kinases AtMPK6 and AtMPK3. It also constitutively expresses a variety of defense-associated genes, including those encoding the WRKY transcription factors AtWRKY29 and AtWRKY6, the MAP kinases AtMPK6 and AtMPK3, the powdery mildew R proteins RPW8.1 and RPW8.2, EDS1 and PR proteins. All of these ssi4-induced responses, as well as the chlorotic, stunted morphology and enhanced disease resistance phenotype, are suppressed by high RH (95%) growth conditions. Thus, a humidity sensitive factor (HSF) appears to function at an early point in the ssi4 signaling pathway. All ssi4 phenotypes, except for MAP kinase activation, also were suppressed by the eds1-1 mutation. Thus, ssi4-induced MAP kinase activation occurs downstream of the HSF but either upstream of EDS1 or on a separate branch of the ssi4 signaling pathway. SA is a critical signaling component in ssi4-mediated defense responses. However, exogenously supplied SA failed to restore lesion formation in high RH-grown ssi4 plants, although it induced defense gene expression. Thus, additional signals also are involved. PMID- 15341635 TI - The glycan substrate of the cytosolic (Pho 2) phosphorylase isozyme from Pisum sativum L.: identification, linkage analysis and subcellular localization. AB - The subcellular distribution of starch-related enzymes and the phenotype of Arabidopsis mutants defective in starch degradation suggest that the plastidial starch turnover is linked to a cytosolic glycan metabolism. In this communication, a soluble heteroglycan (SHG) from leaves of Pisum sativum L. has been studied. Major constituents of the SHG are galactose, arabinose and glucose. For subcellular location, the SHG was prepared from isolated protoplasts and chloroplasts. On a chlorophyll basis, protoplasts and chloroplasts yielded approximately 70% and less than 5%, respectively, of the amount of the leaf derived SHG preparation. Thus, most of SHG resides inside the cell but outside the chloroplast. SHG is soluble and not membrane-associated. Using membrane filtration, the SHG was separated into a <10 kDa and a >10 kDa fraction. The latter was resolved into two subfractions (I and II) by field-flow fractionation. In the protoplast-derived >10 kDa SHG preparation the subfraction I was by far the most dominant compound. beta-Glucosyl Yariv reagent was reactive with subfraction II, but not with subfraction I. In in vitro assays the latter acted as glucosyl acceptor for the cytosolic (Pho 2) phosphorylase but not for rabbit muscle phosphorylase. Glycosidic linkage analyses of subfractions I and II and of the Yariv reagent reactive glycans revealed that all three glycans contain a high percentage of arabinogalactan-like linkages. However, SHG possesses a higher content of minor compounds, namely glucosyl, mannosyl, rhamnosyl and fucosyl residues. Based on glycosyl residues and glycosidic linkages, subfraction I possesses a more complex structure than subfraction II. PMID- 15341636 TI - Multiple gene detection by in situ RT-PCR in isolated plant cells and tissues. AB - With the number of functional genomic approaches in plant biology increasing daily, the demand for rapid and reliable RNA localization techniques for gene characterization is being felt. We present herein a novel, liquid phase in situ RT-PCR (IS-RT-PCR) protocol using a combination of gene-specific fluorescent primers and spectral confocal microscopy to localize target RNA in epicotyl sections and xylogenic suspension cultures of Zinnia elegans. Potential sources of artefacts from fixation to gene detection were systematically eliminated using both fluorescent primers and nucleotides for 18S rRNA gene detection, resulting in a set of optimal parameters for IS-RT-PCR that may be readily adapted to any target gene. By judiciously choosing fluorescent primers with non-overlapping fluorochromes, we have shown that our technique is readily adapted to multiplex IS-RT-PCR, enabling the simultaneous localization of more than one gene within a complex tissue or heterogeneous cell population. A 6-carboxy-2',4,4',5',7,7' hexachlorofluorescein (6-HEX)-labelled primer and a tetrachloro-6-carboxy fluorescein (TET)-labelled primer were designed for two marker genes associated with programmed cell death in tracheary elements (TEs): an endonuclease (Zen1) and a cysteine protease (ZcP4), respectively. An additional Cyan5 (Cy5)-labelled primer was used to monitor 18SrRNA expression. As expected, the 18S signal was constitutively expressed throughout epicotyls sections and living cells in xylogenic in vitro cultures, whereas Zen1 and ZcP4 were co-localized in forming TEs both in planta and in vitro. Analogous to clustering analysis of gene expression using microarrays to elucidate common metabolic pathways and developmental processes, this novel technique is perfectly adapted to gaining a better understanding of gene function via the coordinated expression of genes in specific cell types of complex tissues and cell populations. PMID- 15341637 TI - Dissecting large and complex genomes: flow sorting and BAC cloning of individual chromosomes from bread wheat. AB - The analysis of the complex genome of common wheat (Triticum aestivum, 2n = 6x = 42, genome formula AABBDD) is hampered by its large size ( approximately 17 000 Mbp) and allohexaploid nature. In order to simplify its analysis, we developed a generic strategy for dissecting such large and complex genomes into individual chromosomes. Chromosome 3B was successfully sorted by flow cytometry and cloned into a bacterial artificial chromosome (BAC), using only 1.8 million chromosomes and an adapted protocol developed for this purpose. The BAC library (designated as TA-3B) consists of 67 968 clones with an average insert size of 103 kb. It represents 6.2 equivalents of chromosome 3B with 100% coverage and 90% specificity as confirmed by genetic markers. This method was validated using other chromosomes and its broad application and usefulness in facilitating wheat genome analysis were demonstrated by target characterization of the chromosome 3B structure through cytogenetic mapping. This report on the successful cloning of flow-sorted chromosomes into BACs marks the integration of flow cytogenetics and genomics and represents a great leap forward in genetics and genomic analysis. PMID- 15341638 TI - Traffic spotting: poles apart. AB - Finding out where specific functions are carried out within a bacterial cell has now become technically feasible. Here we consider recent experiments aimed at determining where bacteria translocate proteins across the cytoplasmic membrane using the Sec machinery. PMID- 15341639 TI - Quorum sensing and signal interference: diverse implications. AB - Quorum sensing (QS) is a community genetic regulation mechanism that controls microbiological functions of medical, agricultural and industrial importance. Discovery of microbial QS signals and the signalling mechanisms led to identification of numerous enzymatic and non-enzymatic signal interference mechanisms that quench microbial QS signalling. Evidence is accumulating that such signal interference mechanisms can be developed as promising approaches to control microbial infection and biofilm formation. In addition, these mechanisms exist not only in microorganisms but also in the host organisms of bacterial pathogens, highlighting their potential implications in microbial ecology and in host-pathogen interactions. Investigation of QS and signal interference mechanisms might significantly broaden the scope of research in microbiology. PMID- 15341640 TI - Genetic mapping in the human malaria parasite Plasmodium falciparum. AB - The Plasmodium falciparum genome sequence has boosted hopes for a new era of malaria research and for the application of comprehensive molecular knowledge to disease control, but formidable obstacles remain: approximately 60% of the predicted P. falciparum proteins have no known functions or homologues, and most life cycle stages of this haploid eukaryotic parasite are relatively intractable to cultivation and biochemical manipulation. Genetic mapping based on high resolution maps saturated with single-nucleotide polymorphisms or microsatellites is now providing effective strategies for discovering candidate genes determining important parasite phenotypes. Here we review classical linkage studies using laboratory crosses and population associations that are now amenable to genome wide approaches and are revealing multiple candidate genes involved in complex drug responses. Moreover, mapping by linkage disequilibrium is practicable in cases where chromosomal segments flanking drug-selected genes have been preserved in populations during relatively recent P. falciparum evolution. We discuss the advantages and limitations of these various genetic mapping strategies, results from which offer complementary insights to those emerging from gene knockout experiments and/or high-throughput genomic technologies. PMID- 15341641 TI - Subcellular sites for bacterial protein export. AB - Most bacterial proteins destined to leave the cytoplasm are exported to extracellular compartments or imported into the cytoplasmic membrane via the highly conserved SecA-YEG pathway. In the present studies, the subcellular distributions of core components of this pathway, SecA and SecY, and of the secretory protein pre-AmyQ, were analysed using green fluorescent protein fusions, immunostaining and/or immunogold labelling techniques. It is shown that SecA, SecY and (pre-)AmyQ are located at specific sites near and/or in the cytoplasmic membrane of Bacillus subtilis. The localization patterns of these proteins suggest that the Sec machinery is organized in spiral-like structures along the cell, with most of the translocases organized in specific clusters along these structures. However, this localization appears to be independent of the helicoidal structures formed by the actin-like cytoskeletal proteins, MreB or Mbl. Interestingly, the specific localization of SecA is dynamic, and depends on active translation. Moreover, reducing the phosphatidylglycerol phospholipids content in the bacterial membrane results in delocalization of SecA, suggesting the involvement of membrane phospholipids in the localization process. These data show for the first time that, in contrast to the recently reported uni-ExPortal site in the coccoid Streptococcus pyogenes, multiple sites dedicated to protein export are present in the cytoplasmic membrane of rod-shaped B. subtilis. PMID- 15341642 TI - Lack of mismatch correction facilitates genome evolution in mycobacteria. AB - In silico genome sequence analyses suggested that mycobacteria are devoid of the highly conserved mutLS-based post-replicative mismatch repair system. Here, we present the first biological evidence for the lack of a classical mismatch repair function in mycobacteria. We found that frameshifts, but not general mutation rates are unusually high in Mycobacterium smegmatis. However, despite the absence of mismatch correction, M. smegmatis establishes a strong barrier to recombination between homeologous DNA sequences. We show that 10-12% of DNA sequence heterology restricts initiation of recombination but not extension of heteroduplex DNA intermediates. Together, the lack of mismatch correction and a high stringency of initiation of homologous recombination provide an adequate strategy for mycobacterial genome evolution, which occurs by gene duplication and divergent evolution. PMID- 15341643 TI - The gprA and gprB genes encode putative G protein-coupled receptors required for self-fertilization in Aspergillus nidulans. AB - The filamentous fungus Aspergillus nidulans possesses both asexual and sexual reproductive cycles. Sexual fruiting bodies (cleistothecia) can be formed in both homothallic (self) and heterothallic (outcross) conditions. In this study, we characterized two genes, gprA and gprB, that are predicted to encode putative G protein-coupled receptors (GPCRs) similar to fungal pheromone receptors. Deletion (Delta) of gprA or gprB resulted in the production of a few small cleistothecia carrying a reduced number of ascospores, whereas DeltagprADeltagprB eliminated fruiting body formation in homothallic conditions. However, nullifying gprA and/or gprB did not affect vegetative growth, asexual sporulation, Hulle cell formation or even cleistothecia formation in outcross, indicating that GprA and GprB are specifically required for self-fertilization. The gprA and gprB genes encode two transcripts and, for both genes, larger transcripts are detectable during vegetative growth and asexual development whereas smaller transcripts accumulate during sexual development. Upregulation of nsdD encoding a key sexual developmental activator resulted in the production of barren cleistothecia in the DeltagprADeltagprB mutant, suggesting that NsdD can partially rescue the developmental defects caused by deletion of GPCRs and that GprA/B-mediated signalling may activate other genes necessary for maturation of cleistothecia and ascosporogenesis. Deletion of gprA and/or gprB suppressed growth defects caused by DeltagprD, implying that GprA/B function downstream of GprD-mediated negative control of sexual development. PMID- 15341644 TI - Characterization of IDI-4, a bZIP transcription factor inducing autophagy and cell death in the fungus Podospora anserina. AB - In filamentous fungi a cell death reaction occurs when hyphae of unlike genotype fuse. This phenomenon is referred to as heterokaryon incompatibility. In Podospora anserina, this cell death reaction was found to be associated with the transcriptional induction of a set of genes termed idi genes (for induced during incompatibility) and activation of autophagy. Herein, we describe the characterization of idi-4, a novel idi gene encoding a bZIP transcription factor. Expression of idi-4 is induced during cell death by incompatibility and in various stress conditions. Inactivation of idi-4 by gene replacement does not suppress incompatibility but we show that overexpression of idi-4 triggers cell death. Strains which undergo idi-4-induced cell death display cytological hallmarks of cell death by incompatibility notably induction of autophagy. We also report that increased expression of idi-4 leads to transcriptional induction of other idi genes such as idi-7, the orthologue of the yeast ATG8 autophagy gene. Together these results establish IDI-4 as one of the transcription factor regulating autophagy and cell fate in Podospora. PMID- 15341645 TI - Ralstonia solanacearum genes induced during growth in tomato: an inside view of bacterial wilt. AB - The phytopathogen Ralstonia solanacearum has over 5000 genes, many of which probably facilitate bacterial wilt disease development. Using in vivo expression technology (IVET), we screened a library of 133 200 R. solanacearum strain K60 promoter fusions and isolated approximately 900 fusions expressed during bacterial growth in tomato plants. Sequence analysis of 307 fusions revealed 153 unique in planta-expressed (ipx) genes. These genes included seven previously identified virulence genes (pehR, vsrB, vsrD, rpoS, hrcC, pme and gspK) as well as seven additional putative virulence factors. A significant number of ipx genes may reflect adaptation to the host xylem environment; 19.6%ipx genes are predicted to encode proteins with metabolic and/or transport functions, and 9.8%ipx genes encode proteins possibly involved in stress responses. Many ipx genes (18%) encode putative transmembrane proteins. A majority of ipx genes isolated encode proteins of unknown function, and 13% were unique to R. solanacearum. The ipx genes were variably induced in planta; beta-glucuronidase reporter gene expression analysis of a subset of 44 ipx fusions revealed that in planta expression levels were between two- and 37-fold higher than in culture. The expression of many ipx genes was subject to known R. solanacearum virulence regulators. Of 32 fusions tested, 28 were affected by at least one virulence regulator; several fusions were controlled by multiple regulators. Two ipx fusion strains isolated in this screen were reduced in virulence on tomato, indicating that gene(s) important for bacterial wilt pathogenesis were interrupted by the IVET insertion; mutations in other ipx genes are necessary to determine their roles in virulence and in planta growth. Collectively, this profile of ipx genes suggests that in its host, R. solanacearum confronts and overcomes a stressful and nutrient-poor environment. PMID- 15341646 TI - Deletion of the Gibberella fujikuroi glutamine synthetase gene has significant impact on transcriptional control of primary and secondary metabolism. AB - In Gibberella fujikuroi, the gibberellin (GA) and bikaverin biosynthesis are under control of nitrogen metabolite repression. However, the signalling components acting upstream of AREA are still unknown. We investigated the role of glutamine synthetase (GS) both as an enzyme and as a possible regulator in the nitrogen regulation system. We cloned and replaced the GS-encoding gene, glnA-Gf. The mutants grow with a phenotype different from the wild type in the presence of glutamine. They were unable to express nitrogen-repressed GA and bikaverin biosynthetic genes even under nitrogen starvation conditions. Complementation with the glnA-Gf wild-type copy fully restored GS activity, the expression of secondary metabolism genes, and the production of GAs and the red pigment, bikaverin. In order to find more target genes of GS, differential cDNA-screening and differential hybridization of macroarrays were performed using cDNA from the wild type and DeltaglnA mutant as probes. Several genes were dramatically up- or downregulated in the mutant. Among them are genes involved in N- and C catabolism, and in transcriptional and translation control. Some of these genes are also under AREA control. Treatment with the GS inhibitor l-methionine sulphoximine resulted in similar expression patterns as in the glnA mutant with ammonium as nitrogen source, whereas glutamine can overcome the up- or downregulation of most but not all of the target genes. These findings suggest that not only glutamine, but also GS itself might play an important role in nitrogen metabolite repression. PMID- 15341647 TI - A mycobacterial virulence gene cluster extending RD1 is required for cytolysis, bacterial spreading and ESAT-6 secretion. AB - Initiation and maintenance of infection by mycobacteria in susceptible hosts are not well understood. A screen of Mycobacterium marinum transposon mutant library led to isolation of eight mutants that failed to cause haemolysis, all of which had transposon insertions in genes homologous to a region between Rv3866 and Rv3881c in Mycobacterium tuberculosis, which encompasses RD1 (Rv3871-Rv3879c), a known virulence gene cluster. The M. marinum mutants showed decreased virulence in vivo and failed to secrete ESAT-6, like M. tuberculosis RD1 mutants. M. marinum mutants in genes homologous to Rv3866-Rv3868 also failed to accumulate intracellular ESAT-6, suggesting a possible role for those genes in synthesis or stability of the protein. These transposon mutants and an ESAT-6/CFP-10 deletion mutant all showed reduced cytolysis and cytotoxicity to macrophages and significantly decreased intracellular growth at late stages of the infection only when the cells were infected at low multiplicity of infection, suggesting a defect in spreading. Direct evidence for cell-to-cell spread by wild-type M. marinum was obtained by microscopic detection in macrophage and epithelial monolayers, but the mutants all were defective in this assay. Expression of M. tuberculosis homologues complemented the corresponding M. marinum mutants, emphasizing the functional similarities between M. tuberculosis and M. marinum genes in this region that we designate extRD1 (extended RD1). We suggest that diminished membranolytic activity and defective spreading is a mechanism for the attenuation of the extRD1 mutants. These results extend recent findings on the genomic boundaries and functions of M. tuberculosis RD1 and establish a molecular cellular basis for the role that extRD1 plays in mycobacterial virulence. Disruption of the M. marinum homologue of Rv3881c, not previously implicated in virulence, led to a much more attenuated phenotype in macrophages and in vivo, suggesting that this gene plays additional roles in M. marinum survival in the host. PMID- 15341648 TI - Independent genetic mechanisms mediate turgor generation and penetration peg formation during plant infection in the rice blast fungus. AB - The first barrier to infection encountered by foliar pathogens is the host cuticle. To traverse this obstacle, many fungi produce specialized infection cells called appressoria. MST12 is essential for appressorium-mediated penetration and infectious growth by the rice pathogen Magnaporthe grisea. In this study, we have characterized in detail the penetration defects of an mst12 deletion mutant. Appressoria formed by the mst12 mutant developed normal turgor pressure and ultrastructure but failed to form penetration pegs either on cellophane membranes or on plant epidermal cells. Deletion and site-directed mutagenesis analyses indicated that both the homeodomain and zinc finger domains, but not the middle region, of MST12 are essential for appressorial penetration and plant infection. The mst12 mutant appeared to be defective in microtubule reorganization associated with penetration peg formation. In mature appressoria, the mutant lacked vertical microtubules observed in the wild type. The mst12 mutant also failed to elicit localized host defence responses, including papilla formation and autofluorescence. Our data indicate that generation of appressorium turgor pressure and formation of the penetration peg are two independent processes. MST12 may play important roles in regulating penetration peg formation and directing the physical forces exerted by the appressorium turgor in mature appressoria. PMID- 15341649 TI - Newly secreted adenylate cyclase toxin is responsible for intoxication of target cells by Bordetella pertussis. AB - Adenylate cyclase (AC) toxin is present on the surface of Bordetella pertussis organisms and their addition to eukaryotic cells results in increases in intracellular cAMP. To test the hypothesis that surface-bound toxin is the source for intoxication of cells when incubated with B. pertussis, we characterized the requirements of intoxication from intact bacteria and found that this process is calcium-dependent and blocked by monoclonal antibody to AC toxin or antibody against CD11b, a surface glycoprotein receptor for the toxin. Increases in intracellular cAMP correlate with the number of adherent bacteria, not the total number present in the medium, suggesting that interaction of bacteria with target cells is important for efficient delivery of AC toxin. A filamentous haemagglutinin-deficient mutant (BP353) and a clinical isolate (GMT1), both of which have a marked reduction in AC toxin on their surface, and wild-type B. pertussis (BP338) from which surface AC toxin has been removed by trypsin, were fully competent for intoxicating target cells, demonstrating that surface-bound AC toxin is not responsible for intoxication. B. pertussis killed by gentamicin or gamma irradiation were unable to intoxicate, illustrating that toxin delivery requires viable bacteria. Furthermore, CCCP, a protonophore that disrupts the proton gradient necessary for the secretion of related RTX toxins, blocked intoxication by whole bacteria. These data establish that delivery of this toxin by intact B. pertussis is not dependent on the surface-associated AC toxin, but requires close association of live bacteria with target cells and the active secretion of AC toxin. PMID- 15341650 TI - Involvement of an X family DNA polymerase in double-stranded break repair in the radioresistant organism Deinococcus radiodurans. AB - DNA polymerases of the X family have been implicated in a variety of DNA repair processes in eukaryotes. Here we show that Deinococcus radiodurans, a highly radioresistant bacterium able to mend hundreds of radiation-induced double stranded DNA breaks, expresses a DNA polymerase belonging to the X family. This novel bacterial polymerase, named PolX(Dr), was identified as the product of the Deinococcal DR0467 gene. The purified PolX(Dr) protein possesses a DNA polymerase activity that is stimulated by MnCl2, a property of the X family DNA polymerases. Antibodies raised against PolX(Dr) recognized human pol lambda, rat pol beta and yeast Pol4 and, conversely, antibodies raised against these proteins recognized PolX(Dr). This immunological cross-reactivity suggests a high degree of structural conservation among the polymerases of the X family. Lack of PolX(Dr) reduced the rate of repair of double-stranded DNA breaks and increased cell sensitivity to gamma-rays. PolX(Dr) thus appears to play an important role in double-stranded DNA break repair in D. radiodurans. PMID- 15341651 TI - Endoplasmic reticulum stress leads to the selective transcriptional downregulation of the glucoamylase gene in Aspergillus niger. AB - We describe a new endoplasmic reticulum (ER)-associated stress response in the filamentous fungus Aspergillus niger. The inhibition of protein folding within the ER leads to cellular responses known collectively as the unfolded protein response (UPR) and we show that the selective transcriptional downregulation of the gene encoding glucoamylase, a major secreted protein, but not two non secreted proteins, is an additional consequence of ER stress. The transcriptional downregulation effect is shown by nuclear run-on studies to be at the level of transcription, rather than mRNA stability, and is found to be mediated through the promoter of glaA in a region more than 1 kb upstream of the translational start. The inhibition of protein folding in the ER can be induced in a variety of ways. We examined the effects of dithiothreitol (DTT), a reducing agent that causes the formation of unfolded proteins. Although a general downregulation of transcription was seen with DTT treatment, we show that selective downregulation was observed with the glaA gene compared with genes encoding the non-secreted proteins gamma-actin and glyceraldehyde 3'-phosphate dehydrogenase. The DTT treated fungal cells also showed evidence for the induction of the UPR because expression of bipA and pdiA, encoding an ER-resident chaperone and foldase, respectively, are upregulated and splicing of hacA, the gene encoding the transcription factor responsible for induction of the UPR, occurs allowing the production of an active HacA protein. As a preliminary attempt to investigate if the transcriptional downregulation effect was mediated through HacA (i.e. part of the UPR), we examined ER stress induced through antisense technology to lower the level of PDI in the ER of A. niger. Although the transcription of glaA was attenuated in that strain of A. niger, UPR was not evident, suggesting that the transcriptional downregulation mechanism is controlled differently from the UPR. PMID- 15341652 TI - Rck1 and Rck2 MAPKAP kinases and the HOG pathway are required for oxidative stress resistance. AB - We demonstrate a role in oxidative and metal stress resistance for the MAPK activated protein kinases Rck1 and Rck2 in Saccharomyces cerevisiae. We show that Hog1 is robustly phosphorylated in a Pbs2-dependent way during oxidative stress, and that Rck2 also is phosphorylated under these circumstances. Hog1 concentrates in the nucleus in oxidative stress. Hog1 localization is partially dependent on Rck2, as rck2 cells have more nuclear Hog1 than wild-type cells. We find several proteins with a role in oxidative stress resistance using Rck1 or Rck2 as baits in a two-hybrid screen. We identify the transcription factor Yap2 as a putative target for Rck1, and the Zn2+ transporter Zrc1 as a target for Rck2. Yap2 is normally cytoplasmic, but rapidly migrates to the nucleus upon exposure to oxidative stress agents. In a fraction of untreated pbs2 cells, Yap2 is nuclear. Zrc1 co-immunoprecipitates with Rck2, and ZRC1 is genetically downstream of RCK2. These data connect activation of the Hog1 MAPK cascade with effectors having a role in oxidative stress resistance. PMID- 15341653 TI - Identification of a gene required for the formation of lyso-ornithine lipid, an intermediate in the biosynthesis of ornithine-containing lipids. AB - Under phosphate-limiting conditions, some bacteria replace their membrane phospholipids by lipids not containing any phosphorus. One of these phosphorus free lipids is an ornithine-containing lipid (OL) that is widespread among eubacteria. In earlier work, we had identified a gene (olsA) required for OL biosynthesis that probably encodes an O-acyltransferase using acyl-acyl carrier protein (acyl-AcpP) as an acyl donor and that converts lyso-ornithine lipid into OL. We now report on a second gene (olsB) required for OL biosynthesis that is needed for the incorporation of radiolabelled ornithine into OL. Overexpression of OlsB in an olsA-deficient mutant of Sinorhizobium (Rhizobium) meliloti leads to the transient accumulation of lyso-ornithine lipid, the biosynthetic intermediate of OL biosynthesis. Overexpression of OlsB in Escherichia coli is sufficient to cause the in vivo formation of lyso-ornithine lipid in this organism and is the cause for a 3-hydroxyacyl-AcpP-dependent acyltransferase activity forming lyso-ornithine lipid from ornithine. These results demonstrate that OlsB is required for the first step of OL biosynthesis, in which ornithine is N-acylated with a 3-hydroxy-fatty acyl residue in order to obtain lyso ornithine lipid. OL formation in a wild-type S. meliloti is increased upon growth under phosphate-limiting conditions. Expression of OlsB from a broad host range vector leads to the constitutive formation of relatively high amounts of OL (12 14% of total membrane lipids) independently of whether strains are grown in the presence of low or high concentrations of phosphate, suggesting that in S. meliloti the formation of OlsB is usually limiting for the amount of OL formed in this organism. Open reading frames homologous to OlsA and OlsB were identified in many eubacteria and although in S. meliloti the olsB and olsA gene are 14 kb apart, in numerous other bacteria they form an operon. PMID- 15341654 TI - Complex spatial distribution and dynamics of an abundant Escherichia coli outer membrane protein, LamB. AB - Advanced techniques for observing protein localization in live bacteria show that the distributions are dynamic. For technical reasons, most such techniques have not been applied to outer membrane proteins in Gram-negative bacteria. We have developed two novel live-cell imaging techniques to observe the surface distribution of LamB, an abundant integral outer membrane protein in Escherichia coli responsible for maltose uptake and for attachment of bacteriophage lambda. Using fluorescently labelled bacteriophage lambda tails, we quantitatively described the spatial distribution and dynamic movement of LamB in the outer membrane. LamB accumulated in spiral patterns. The distribution depended on cell length and changed rapidly. The majority of the protein diffused along spirals extending across the cell body. Tracking single particles, we found that there are two populations of LamB--one shows very restricted diffusion and the other shows greater mobility. The presence of two populations recalls the partitioning of eukaryotic membrane proteins between 'mobile' and 'immobile' populations. In this study, we have demonstrated that LamB moves along the bacterial surface and that these movements are restricted by an underlying dynamic spiral pattern. PMID- 15341655 TI - Fungicide activity through activation of a fungal signalling pathway. AB - Fungicides generally inhibit enzymatic reactions involved in fungal cellular biosynthesis. Here we report, for the first time, an example of fungicidal effects through hyperactivation of a fungal signal transduction pathway. The OSC1 gene, encoding a MAP kinase (MAPK) related to yeast Hog1, was isolated from the fungal pathogen Colletotrichum lagenarium that causes cucumber anthracnose. The osc1 knockout mutants were sensitive to high osmotic stress and showed increased resistance to the fungicide fludioxonil, indicating that Osc1 is involved in responses to hyperosmotic stress and sensitivity to fludioxonil. The Osc1 MAPK is phosphorylated under high osmotic conditions, indicating activation of Osc1 by high osmotic stress. Importantly, fludioxonil treatment also activates phosphorylation of Osc1, suggesting that improper activation of Osc1 by fludioxonil has negative effects on fungal growth. In the presence of fludioxonil, the wild-type fungus was not able to infect the host plant because of a failure of appressorium-mediated penetration, whereas osc1 mutants successfully infected plants. Analysis using a OSC1-GFP fusion gene indicated that Osc1 is rapidly translocated to the nucleus in appressorial cells after the addition of fludioxonil, suggesting that fludioxonil impairs the function of infection structures by activation of Osc1. Furthermore, fludioxonil activates Hog1-type MAPKs in the plant pathogenic fungi Cochliobolus heterostrophus and Botrytis cinerea. These results strongly suggest that fludioxonil acts as a fungicide, in part, through activation of the MAPK cascade in fungal pathogens. PMID- 15341656 TI - Predictors of vigorous exercise adoption and maintenance over four years in a community sample. AB - BACKGROUND: Very little is known about the correlates of adoption and maintenance of vigorous exercise. The purpose of this study was to understand the sociodemographic correlates of exercise adoption and maintenance in a community sample. METHODS: 917 women and 229 men completed annual surveys as part of a community-based weight gain prevention trial over four years. Multivariate regressions evaluated predictive factors for maintenance of vigorous exercise over time in regular exercisers, and predictors of adoption of exercise in adults who were sedentary at baseline. RESULTS: Exercise maintenance at Years 2 and 3 was associated with ethnicity and exercise level at baseline, while exercise maintenance at Year 4 was associated with television watching, BMI and exercise at baseline. Exercise level at baseline was associated with exercise initiation at Year 2 and Year 3. Income level, marital status, and smoking status predicted exercise initiation at Year 4. CONCLUSIONS: Predictors of vigorous exercise maintenance were more consistent than predictors of vigorous exercise initiation. Results suggest that those who adopt vigorous exercise are a heterogeneous group and intervention messages could be more broadly focused. These data also suggest that exercise maintenance interventions should continue to target low-income populations with messages regarding smoking, weight and television. Clearly further research is needed to understand the factors that contribute to exercise initiation and maintenance, and to develop effective interventions to improve levels of physical activity levels. PMID- 15341657 TI - Comparison of the NEI-VFQ and OSDI questionnaires in patients with Sjogren's syndrome-related dry eye. AB - BACKGROUND: To examine the associations between vision-targeted health-related quality of life (VT-HRQ) and ocular surface parameters in patients with Sjogren's syndrome, a systemic autoimmune disease characterized by dry eye and dry mouth. METHODS: Forty-two patients fulfilling European / American diagnostic criteria for Sjogren's syndrome underwent Schirmer testing without anesthesia, ocular surface vital dye staining; and measurement of tear film breakup time (TBUT). Subjects were administered the Ocular Surface Disease Index (OSDI) and the 25 item National Eye Institute Vision Functioning Questionnaire (NEI-VFQ). Main outcome measures included ocular surface parameters, OSDI subscales describing ocular discomfort (OSDI-symptoms), vision-related function (OSDI-function), and environmental triggers, and NEI-VFQ subscales. RESULTS: Participants (aged 31-81 y; 95% female) all had moderate to severe dry eye. Associations of OSDI subscales with the ocular parameters were modest (Spearman r (rho) < 0.22) and not statistically significant. Associations of NEI-VFQ subscales with the ocular parameters reached borderline significance for the near vision subscale with TBUT (rho = 0.32, p =.05) and for the distance vision subscale with van Bijsterveld score (rho = 0.33, p =.04). The strongest associations of the two questionnaires were for: ocular pain and mental function with OSDI-symptoms (rho = 0.60 and 0.45, respectively); and general vision, ocular pain, mental function, role function, and driving with OSDI-function (rho = 0.60, 0.50, 0.61, 0.64, 0.57, and 0.67, respectively). CONCLUSIONS: Associations between conventional objective measures of dry eye and VT-HRQ were modest. The generic NEI-VFQ was similar to the disease-specific OSDI in its ability to measure the impact of Sjogren's syndrome-related dry eye on VT-HRQ. PMID- 15341658 TI - Characterization of the microheterogeneity of transthyretin in plasma and urine using SELDI-TOF-MS immunoassay. AB - BACKGROUND: It has been shown that transthyretin (TTR) exists in different molecular variants. Besides point mutations associated with different diseases such as amyloidosis, other posttranslational modifications occur that might be of diagnostic interest. RESULTS: TTR levels as determined by ELISA in plasma and urine of healthy individuals were 489 +/- 155 microg/ml plasma and 46 +/- 24 ng/g creatinine, respectively. Average levels in urine of pregnant women were 45 +/- 65 microg/g creatinine. The molecular heterogeneity of TTR was analyzed using a high-throughput mass spectrometric immunoassay system. TTR was extracted from plasma or urine onto an antibody-coated (via protein A) affinity chip surface (PS20) using the surface-enhanced laser desorption/ionization (SELDI) technique. Subsequently samples were subjected to time-of-flight mass spectrometry (TOF-MS). In healthy individuals, TTR in plasma occurred rather consistently in two variants of 13732 +/- 12 and 13851 +/- 9 Da for the native and S-cysteinylated forms and at a smaller signal of 14043 +/- 17 Da for the S-glutathionylated form. In urine of pregnant women, various signals were observed with a dominant signal at 13736 +/- 10 Da and a varying number of smaller immunoreactive fragments. These fragments are possibly the consequence of metabolism in plasma or kidney. CONCLUSION: This chip-based approach represents a rapid and accurate method to characterize the molecular variants of TTR including protein or peptide fragments which are either related to TTR or have resulted from its catabolism. These molecular variants may be of diagnostic importance as alternative or novel biomarkers due to their predominant relation to the TTR metabolism both in healthy and diseased individuals. PMID- 15341659 TI - Simultaneous suppression of disturbing fields and localization of magnetic markers by means of multipole expansion. AB - BACKGROUND: Magnetically marked capsules serve for the analysis of peristalsis and throughput times within the intestinal tract. Moreover, they can be used for the targeted disposal of drugs. The capsules get localized in time by field measurements with a superconducting quantum interference device (SQUID) magnetometer array. Here it is important to ensure an online localization with high speed and high suppression of disturbing fields. In this article we use multipole expansions for the simultaneous localization and suppression of disturbing fields. METHODS: We expand the measurement data in terms of inner and outer multipoles. Thereby we obtain directly a separation of marker field and outer disturbing fields. From the inner dipoles and quadrupoles we compute the magnetization and position of the capsule. The outer multipoles get eliminated. RESULTS: The localization goodness has been analyzed depending on the order of the multipoles used and depending on the systems noise level. We found upper limits of the noise level for the usage of certain multipole moments. Given a signal to noise ratio of 40 and utilizing inner dipoles and quadrupoles and outer dipoles, the method enables an accuracy of 5 mm with a speed of 10 localizations per second. CONCLUSION: The multipole localization is an effective method and is capable of online-tracking magnetic markers. PMID- 15341660 TI - Liver sinusoidal endothelial cell modulation upon resection and shear stress in vitro. AB - BACKGROUND: Shear stress forces acting on liver sinusoidal endothelial cells following resection have been noted as a possible trigger in the early stages of hepatic regeneration. Thus, the morphology and gene expression of endothelial cells following partial hepatectomy or shear stress in vitro was studied. RESULTS: Following partial hepatectomy blood flow-to-liver mass ratio reached maximal values 24 hrs post resection. Concomitantly, large fenestrae (gaps) were noted. Exposure of liver sinusoidal endothelial cells, in vitro, to physiological laminar shear stress forces was associated with translocation of vascular endothelial cell growth factor receptor-2 (VEGFR-2) and neuropilin-1 from perinuclear and faint cytoplasmic distribution to plasma membrane and cytoskeletal localization. Under these conditions, VEGFR-2 co-stains with VE cadherin. Unlike VEGFR-2, the nuclear localization of VEGFR-1 was not affected by shear stress. Quantification of the above receptors showed a significant increase in VEGFR-1, VEGFR-2 and neuropilin-1 mRNA following shear stress. CONCLUSION: Our data suggest a possible relation between elevated blood flow associated with partial hepatectomy and the early events occurring thereby. PMID- 15341661 TI - Garlic's ability to prevent in vitro Cu2+-induced lipoprotein oxidation in human serum is preserved in heated garlic: effect unrelated to Cu2+-chelation. AB - BACKGROUND: It has been shown that several extracts and compounds derived from garlic are able to inhibit Cu2+-induced low density lipoprotein oxidation. In this work we explored if the ability of aqueous garlic extract to prevent in vitro Cu2+-induced lipoprotein oxidation in human serum is affected by heating (a) aqueous garlic extracts or (b) garlic cloves. In the first case, aqueous extract of raw garlic and garlic powder were studied. In the second case, aqueous extract of boiled garlic cloves, microwave-treated garlic cloves, and pickled garlic were studied. It was also studied if the above mentioned preparations were able to chelate Cu2+. METHODS: Cu2+-induced lipoprotein oxidation in human serum was followed by the formation of conjugated dienes at 234 nm and 37 degrees C by 240 min in a phosphate buffer 20 mM, pH 7.4. Blood serum and CuSO4 were added to a final concentration of 0.67% and 0.0125 mM, respectively. The lag time and the area under the curve from the oxidation curves were obtained. The Cu2+-chelating properties of garlic extracts were assessed using an approach based upon restoring the activity of xanthine oxidase inhibited in the presence of 0.050 mM Cu2+. The activity of xanthine oxidase was assessed by monitoring the production of superoxide anion at 560 nm and the formation of uric acid at 295 nm. Data were compared by parametric or non-parametric analysis of variance followed by a post hoc test. RESULTS: Extracts from garlic powder and raw garlic inhibited in a dose dependent way Cu2+-induced lipoprotein oxidation. The heating of garlic extracts or garlic cloves was unable to alter significantly the increase in lag time and the decrease in the area under the curve observed with the unheated garlic extracts or raw garlic. In addition, it was found that the garlic extracts were unable to chelate Cu2+. CONCLUSIONS: (a) the heating of aqueous extracts of raw garlic or garlic powder or the heating of garlic cloves by boiling, microwave or pickling do not affect garlic's ability to inhibit Cu2+-induced lipoprotein oxidation in human serum, and (b) this ability is not secondary to Cu2+ chelation. PMID- 15341662 TI - Comparison of Crocus sativus L. and imipramine in the treatment of mild to moderate depression: a pilot double-blind randomized trial [ISRCTN45683816]. AB - BACKGROUND: The morbidity and mortality associated with depression are considerable and continue to increase. Depression currently ranks fourth among the major causes of disability worldwide, after lower respiratory infections, prenatal conditions, and HIV/AIDS. Crocus sativus L. is used to treat depression. Many medicinal plants textbooks refer to this indication whereas there is no evidence-based document. Our objective was to compare the efficacy of stigmas of Crocus sativus (saffron) with imipramine in the treatment of mild to moderate depression in a 6-week pilot double-blind randomized trial. METHODS: Thirty adult outpatients who met the Diagnostic and Statistical Manual of Mental Disorders, 4th edition for major depression based on the structured clinical interview for DSM IV participated in the trial. Patients have a baseline Hamilton Rating Scale for Depression score of at least 18. In this double-blind, single-center trial, patients were randomly assigned to receive capsule of saffron 30 mg/day (TDS) (Group 1) and capsule of imipramine 100 mg/day (TDS) (Group 2) for a 6-week study. RESULTS: Saffron at this dose was found to be effective similar to imipramine in the treatment of mild to moderate depression (F = 2.91, d.f. = 1, P = 0.09). In the imipramine group anticholinergic effects such as dry mouth and also sedation were observed more often that was predictable. CONCLUSION: The main overall finding from this study is that saffron may be of therapeutic benefit in the treatment of mild to moderate depression. To the best of our knowledge this is the first clinical trial that supports this indication for saffron. A large scale trial with placebo control is warranted. PMID- 15341663 TI - Rapid and reliable extraction of genomic DNA from various wild-type and transgenic plants. AB - BACKGROUND: DNA extraction methods for PCR-quality DNA from calluses and plants are not time efficient, since they require that the tissues be ground in liquid nitrogen, followed by precipitation of the DNA pellet in ethanol, washing and drying the pellet, etc. The need for a rapid and simple procedure is urgent, especially when hundreds of samples need to be analyzed. Here, we describe a simple and efficient method of isolating high-quality genomic DNA for PCR amplification and enzyme digestion from calluses, various wild-type and transgenic plants. RESULTS: We developed new rapid and reliable genomic DNA extraction method. With our developed method, plant genomic DNA extraction could be performed within 30 min. The method was as follows. Plant tissue was homogenized with salt DNA extraction buffer using hand-operated homogenizer and extracted by phenol:chloroform:isoamyl alcohol (25:24:1). After centrifugation, the supernatant was directly used for DNA template for PCR, resulting in successful amplification for RAPD from various sources of plants and specific foreign genes from transgenic plants. After precipitating the supernatant, the DNA was completely digested by restriction enzymes. CONCLUSION: This DNA extraction procedure promises simplicity, speed, and efficiency, both in terms of time and the amount of plant sample required. In addition, this method does not require expensive facilities for plant genomic DNA extraction. PMID- 15341664 TI - Multi-line split DNA synthesis: a novel combinatorial method to make high quality peptide libraries. AB - BACKGROUND: We developed a method to make a various high quality random peptide libraries for evolutionary protein engineering based on a combinatorial DNA synthesis. RESULTS: A split synthesis in codon units was performed with mixtures of bases optimally designed by using a Genetic Algorithm program. It required only standard DNA synthetic reagents and standard DNA synthesizers in three lines. This multi-line split DNA synthesis (MLSDS) is simply realized by adding a mix-and-split process to normal DNA synthesis protocol. Superiority of MLSDS method over other methods was shown. We demonstrated the synthesis of oligonucleotide libraries with 1016 diversity, and the construction of a library with random sequence coding 120 amino acids containing few stop codons. CONCLUSIONS: Owing to the flexibility of the MLSDS method, it will be able to design various "rational" libraries by using bioinformatics databases. PMID- 15341665 TI - An outbreak of Salmonella enteritidis phage type 34a infection associated with a Chinese restaurant in Suffolk, United Kingdom. AB - BACKGROUND: On 30th July 2002, the Suffolk Communicable Disease Control Team received notifications of gastrointestinal illness due to Salmonella Enteritidis in subjects who had eaten food from a Chinese restaurant on 27th July. An Outbreak Control Team was formed resulting in extensive epidemiological, microbiological and environmental investigations. METHODS: Attempts were made to contact everybody who ate food from the restaurant on 27th July and a standard case definition was adopted. Using a pre-designed proforma information was gathered from both sick and well subjects. Food specific attack rates were calculated and two-tailed Fisher's exact test was used to test the difference between type of food consumed and the health status. Using a retrospective cohort design univariate Relative Risks and 95% Confidence Intervals were calculated for specific food items. RESULTS: Data was gathered on 52 people of whom 38 developed gastrointestinal symptoms; 16 male and 22 female. The mean age was 27 years. The mean incubation period was 30 hours with a range of 6 to 90 hours. Food attack rates were significantly higher for egg, special and chicken fried rice. Relative risk and the Confidence interval for these food items were 1.97 (1.11-3.48), 1.56 (1.23-1.97) and 1.48 (1.20-1.83) respectively. Interviews with the chef revealed that many eggs were used in the preparation of egg-fried rice, which was left at room temperature for seven hours and was used in the preparation of the other two rice dishes. Of the 31 submitted stool specimens 28 tested positive for S Enteritidis phage type 34a and one for S Enteritidis phage type 4. CONCLUSION: In the absence of left over food available for microbiological examination, epidemiological investigation strongly suggested the eggs used in the preparation of the egg-fried rice as the vehicle for this outbreak. This investigation highlights the importance of safe practices in cooking and handling of eggs in restaurants. PMID- 15341666 TI - The Internet and HIV study: design and methods. AB - BACKGROUND: The Internet provides a new meeting ground, especially for gay men, that did not exist in the early 1990s. Several studies have found increased levels of high risk sexual behaviour and sexually transmissible infections (STI) among gay men who seek sex on the Internet, although the underlying processes are not fully understood. Research funded by the UK Medical Research Council (2002 2004) provided the opportunity to consider whether the Internet represents a new sexual risk environment for gay and bisexual men living in London. METHODS: The objectives of the Internet and HIV study are to: (i) measure the extent to which gay men living in London seek sexual partners on the Internet; (ii) compare the characteristics of London gay men who do and do not seek sex on the Internet; (iii) examine whether sex with Internet-partners is less safe than with other sexual partners; (iv) compare use of the Internet with other venues where men meet sexual partners; (v) establish whether gay men use the Internet to actively seek partners for unprotected anal intercourse; (vi) determine the potential for using the Internet for HIV prevention. These objectives have been explored using quantitative and qualitative research methods in four samples of London gay men recruited and interviewed both online and offline. The four samples were: (i) gay men recruited through Internet chat rooms and profiles; (ii) HIV positive gay men attending an NHS hospital outpatients clinic; (iii) gay men seeking an HIV test in an NHS HIV testing or sexual health clinic; (iv) gay men recruited in the community. RESULTS: Quantitative data were collected by means of confidential, anonymous self-administered questionnaires (n>4000) completed on-line by the Internet sample. Qualitative data were collected by means of one-to-one interviews (n = 128) conducted either face-to-face or on-line. CONCLUSION: The strength of the Internet and HIV study is its methodological plurality, drawing on both qualitative and quantitative research among online and offline samples, as well as taking advantage of recent advances in web survey design. The study's findings will help us better understand the role of the Internet in relation to gay men's sexual practice. PMID- 15341667 TI - Glutamine-enriched enteral nutrition in very low birth weight infants. Design of a double-blind randomised controlled trial [ISRCTN73254583]. AB - BACKGROUND: Enteral feeding of very low birth weight (VLBW) infants is a challenge, since metabolic demands are high and administration of enteral nutrition is limited by immaturity of the gastrointestinal tract. The amino acid glutamine plays an important role in maintaining functional integrity of the gut. In addition, glutamine is utilised at a high rate by cells of the immune system. In critically ill patients, glutamine is considered a conditionally essential amino acid. VLBW infants may be especially susceptible to glutamine depletion as nutritional supply of glutamine is limited in the first weeks after birth. Glutamine depletion has negative effects on functional integrity of the gut and leads to immunosuppression. This double-blind randomised controlled trial is designed to investigate the effect of glutamine-enriched enteral nutrition on feeding tolerance, infectious morbidity and short-term outcome in VLBW infants. Furthermore, an attempt is made to elucidate the role of glutamine in postnatal adaptation of the gut and modulation of the immune response. METHODS: VLBW infants (gestational age <32 weeks and/or birth weight <1500 g) are randomly allocated to receive enteral glutamine supplementation (0.3 g/kg/day) or isonitrogenous placebo supplementation between day 3 and 30 of life. Primary outcome is time to full enteral feeding (defined as a feeding volume >/= 120 mL/kg/day). Furthermore, incidence of serious infections and short-term outcome are evaluated. The effect of glutamine on postnatal adaptation of the gut is investigated by measuring intestinal permeability and determining faecal microflora. The role of glutamine in modulation of the immune response is investigated by determining plasma Th1/Th2 cytokine concentrations following in vitro whole blood stimulation. PMID- 15341668 TI - HER2 expression in cervical cancer as a potential therapeutic target. AB - BACKGROUND: Trastuzumab, a humanized monoclonal antibody against the HER2 receptor is currently being used in breast and other tumor types. Early studies have shown that a variable proportion of cervical carcinoma tumors overexpress the HER2 receptor as evaluated by diverse techniques and antibodies. Currently it is known that a tumor response to trastuzumab strongly correlates with the level of HER2 expression evaluated by the Hercep Test, thus, it seems desirable to evaluate the status of expression of this receptor using the FDA-approved Hercep Test and grading system to gain insight in the feasibility of using trastuzumab in cervical cancer patients. METHODS: We analyzed a series of cervical cancer cell lines, the primary tumors of 35 cases of cervical cancer patients and four recurrent cases, with the Hercep Test in order to establish whether this tumor type overexpress HER2 at level of 2+/3+ as trastuzumab is currently approved for breast cancer having such level of expression. RESULTS: The results indicate that only 1 out of 35 primary tumors cases overexpress the receptor at this level, however, two out of four recurrent tumors that tested negative at diagnosis shifted to Hercep Test 2+ and 3+ respectively. CONCLUSIONS: The low frequency of expression in primary cases suggests that trastuzumab could have a limited value for the primary management of cervical cancer patients, however, the finding of "conversion" to Hercep Test 2+ and 3+ of recurrent tumors indicates the need to further evaluate the expression of HER2 in the metastatic and recurrent cases. PMID- 15341669 TI - Race/ethnicity and disease severity in IgA nephropathy. AB - BACKGROUND: Relatively few U.S.-based studies in chronic kidney disease have focused on Asian/Pacific Islanders. Clinical reports suggest that Asian/Pacific Islanders are more likely to be affected by IgA nephropathy (IgAN), and that the severity of disease is increased in these populations. METHODS: To explore whether these observations are borne out in a multi-ethnic, tertiary care renal pathology practice, we examined clinical and pathologic data on 298 patients with primary glomerular lesions (IgAN, focal segmental glomerulosclerosis, membranous nephropathy and minimal change disease) at the University of California San Francisco Medical Center from November 1994 through May 2001. Pathologic assessment of native kidney biopsies with IgAN was conducted using Haas' classification system. RESULTS: Among individuals with IgAN (N = 149), 89 (60%) were male, 57 (38%) white, 53 (36%) Asian/Pacific Islander, 29 (19%) Hispanic, 4 (3%) African American and 6 (4%) were of other or unknown ethnicity. The mean age was 37 +/- 14 years and median serum creatinine 1.7 mg/dL. Sixty-six patients (44%) exhibited nephrotic range proteinuria at the time of kidney biopsy. The distributions of age, gender, mean serum creatinine, and presence or absence of nephrotic proteinuria and/or hypertension at the time of kidney biopsy were not significantly different among white, Hispanic, and Asian/Pacific Islander groups. Of the 124 native kidney biopsies with IgAN, 10 (8%) cases were classified into Haas subclass I, 12 (10%) subclass II, 23 (18%) subclass III, 30 (25%) subclass IV, and 49 (40%) subclass V. The distribution of Haas subclass did not differ significantly by race/ethnicity. In comparison, among the random sample of patients with non-IgAN glomerular lesions (N = 149), 77 (52%) patients were male, 51 (34%) white, 42 (28%) Asian/Pacific Islander, 25 (17%) Hispanic, and 30 (20%) were African American. CONCLUSIONS: With the caveats of referral and biopsy biases, the race/ethnicity distribution of IgAN differs significantly from that of other major glomerulonephridities. However, among individuals undergoing native kidney biopsy, we see no evidence of a race/ethnicity association with severity of disease in IgAN by clinical and IgAN-specific histopathologic criteria. Further studies are needed to identify populations at higher risk for progressive disease in IgAN. PMID- 15341670 TI - Antroduodenal motility in neurologically handicapped children with feeding intolerance. AB - BACKGROUND: Dysphagia and feeding intolerance are common in neurologically handicapped children. The aim is to determine the etiologies of feeding intolerance in neurologically handicapped children who are intolerant of tube feedings. METHODS: Eighteen neurologically handicapped children, followed in the Tube Feeding Clinic at the Children's Hospital of Wisconsin who were intolerant of gastrostomy feedings. The charts of these 18 patients were reviewed. Past medical history, diagnoses, history of fundoplication and results of various tests of gastrointestinal function including barium contrast radiography, endoscopy and antroduodenal manometry were documented. RESULTS: Five of 11 children had abnormal barium upper gastrointestinal series. Seven of 14 had abnormal liquid phase gastric emptying tests. Two of 16 had esophagitis on endoscopy. All 18 children had abnormal antroduodenal motility. CONCLUSIONS: In neurologically handicapped children foregut dysmotility may be more common than is generally recognized and can explain many of the upper gastrointestinal symptoms in neurologically handicapped children. PMID- 15341671 TI - Hypoxia upregulates the expression of the NDRG1 gene leading to its overexpression in various human cancers. AB - BACKGROUND: The expression of NDRG1 gene is induced by nickel, a transition metal sharing similar physical properties to cobalt. Nickel may create hypoxia-like conditions in cells and induce hypoxia-responsive genes, as does cobalt. Therefore NDRG1 is likely to be another gene induced by hypoxia. HIF-1 is a transcription factor which has a major role in the regulation of hypoxia responsive genes, and thus it could be involved in the transcriptional regulation of NDRG1 gene. Hypoxia is such a common feature of solid tumours that it is of interest to investigate the expression of Ndrg1 protein in human cancers. RESULTS: Hypoxia and its mimetics induce in vitro expression of NDRG1 gene and cause the accumulation of Ndrg1 protein. Protein levels remain high even after cells revert to normoxia. Although HIF-1 is involved in the regulation of NDRG1, long term hypoxia induces the gene to some extent in HIF-1 knock-out cells. In the majority of human tissues studied, Ndrg1 protein is overexpressed in cancers compared to normal tissues and also reflects tumour hypoxia better than HIF-1 protein. CONCLUSIONS: Hypoxia is an inducer of the NDRG1 gene, and nickel probably causes the induction of the gene by interacting with the oxygen sensory pathway. Hypoxic induction of NDRG1 is mostly dependent on the HIF-1 transcription factor, but HIF-1 independent pathways are also involved in the regulation of the gene during chronic hypoxia. The determination of Ndrg1 protein levels in cancers may aid the diagnosis of the disease. PMID- 15341672 TI - Systemic endopolyploidy in Spathoglottis plicata (Orchidaceae) development. AB - BACKGROUND: Endopolyploidy is developmentally regulated. Presence of endopolyploidy as a result of endoreduplication has been characterized in insects, mammals and plants. The family Orchidaceae is the largest among the flowering plants. Many of the members of the orchid family are commercially micropropagated. Very little has been done to characterize the ploidy variation in different tissues of the orchid plants during development. RESULTS: The DNA contents and ploidy level of nuclei extracted from various tissues of a tropical terrestrial orchid Spathoglottis plicata were examined by flow cytometry. Sepals, petals and ovary tissues were found to have only a 2C (C, DNA content of the unreplicated haploid chromosome complement) peak. Columns, floral pedicels of newly open flowers and growing flower stems were observed to have an endopolyploid 8C peak in addition to 2C and 4C peaks. In developing floral pedicels, four peaks were observed for 2C, 4C, 8C and 16C. In root tips, there were 2C, 4C and 8C peaks. But in the root tissues at the region with root hairs, only a 2C peak was observed. Nuclei extracted from young leaves shown three peaks for 2C, 4C and 8C. A similar pattern was found in the vegetative tissues of both greenhouse-grown plants and tissue-cultured plantlets. In mature leaves, a different pattern of ploidy level was found at different parts of the leaves. In the leaf tips and middle parts, there were 2C and 4C peaks. Only at the basal part of the leaves, there were three peaks for 2C, 4C and 8C. CONCLUSIONS: Systemic variation of cellular endopolyploidy in different tissues during growth and development of Spathoglottis plicata from field-grown plants and in vitro cultures was identified. The implication of the findings was discussed. PMID- 15341673 TI - Neoadjuvant and adjuvant chemotherapy for high-risk localized prostate cancer. AB - The optimal treatment strategy for patients with high-risk localized prostate cancer remains unknown. Definitive local treatments such as radical prostatectomy and external beam radiotherapy cure only a minority of these patients. Recent efforts have been made to reduce the risk of recurrence and delay progression to symptomatic hormone-refractory disease by using chemotherapy before, during, or after definitive local therapy. Chemotherapy is an effective modality in the treatment of hormone- refractory prostate cancer. Studies have established its role in the palliation of symptoms in patients with hormone-refractory disease, though a survival benefit remains to be demonstrated. Prospective randomized trials are underway to test the hypothesis that neoadjuvant and adjuvant chemotherapy may improve survival rate in patients with high-risk localized prostate cancer. The data currently available from nonrandomized trials have not yet established the exact role of neoadjuvant and adjuvant chemotherapy and its potential impact on survival. However, preliminary data suggest that chemotherapy, when administered in concert with definitive local therapy, may be promising in patients with locally advanced prostate cancer. Randomized clinical trials are ongoing to see if neoadjuvant and adjuvant chemotherapy will translate into an improved clinical benefit for the patient, and participation by patients is paramount. We review the recent literature regarding the use of neoadjuvant and adjuvant chemotherapy in patients with locally advanced prostate cancer. PMID- 15341674 TI - Salvage therapy for locally recurrent prostate cancer after external beam radiotherapy. AB - The greatest obstacle in the cure of patients with locally recurrent prostate cancer after radiation therapy is the lack of early detection markers. The majority of patients who are candidates for local salvage therapy have locally advanced disease, precluding successful salvage therapy. A low pretreatment prostate specific antigen (PSA) has shown to be a favorable prognostic variable for disease progression, regardless of the specific local salvage therapy used. Of all the local salvage treatment options for these patients, we believe that salvage radical prostatectomy (RP) offers patients the greatest likelihood of a cure. The salvage RP results approach those achieved with standard RP for patients of similar pathologic stage. When patients are treated early in the course of recurrent disease (preoperative PSA < 10 ng/mL), an estimated two thirds of patients will be disease-free 5 years after salvage RP alone. With better patient selection and continued technical modifications, the morbidity associated with salvage RP has substantially improved. Perioperative complications approach those observed with standard RP and approximately two thirds of patients will recover urinary continence. Select patients may also recover functional erections when nerve-sparing techniques are used. Salvage cryotherapy and brachytherapy are minimally invasive alternatives to salvage RP. The cancer control results of these procedures appear to be inferior to results achieved with salvage RP. Each of these procedures is associated with significant morbidity and do not appear to provide a clear advantage over salvage RP in terms of posttreatment complications, urinary continence, and erectile function. A long term cure is possible for patients with locally recurrent prostate cancer after radiation therapy. Local salvage therapy must be instituted early to be successful in the course of progressive disease. PMID- 15341675 TI - Osteoclast-targeted therapy for prostate cancer. AB - Skeletal complications are a major cause of morbidity in men with metastatic prostate cancer. Bone metastases cause pain, fractures, spinal-cord compression, and ineffective hematopoiesis. Men without bone metastases are also at risk for skeletal complications. Androgen deprivation therapy (ADT), the mainstay of treatment for metastatic prostate cancer and a routine part of the management for many men with nonmetastatic prostate cancer, decreases bone mineral density, and increases fracture risk. Pathological osteoclast activation plays a central role in both disease and treatment-related skeletal morbidity. Bisphosphonates, potent inhibitors of osteoclast activity, are now an important part of the management for many men with prostate cancer. Zoledronic acid, a potent intravenous bisphosphonate, decreases the risk of skeletal complications in men with hormone refractory prostate cancer and bone metastases. Zoledronic acid and pamidronate preserve bone mineral density in men receiving ADT for nonmetastatic prostate cancer. Ongoing clinical trials will evaluate the role of osteoclast-targeted therapy in other settings including prevention of treatment-related fractures, prevention of bone metastases in men with high-risk nonmetastatic prostate cancer, and prevention of skeletal complications in men with hormone-sensitive metastatic disease. PMID- 15341676 TI - Alternatives to cytology in the management of non-muscle invasive bladder cancer. AB - The natural history of non-muscle invasive bladder cancer is characterized by a high probability of recurrence and in the case of high-grade tumors, progression to muscle invasive cancer. This mandates a follow-up strategy designed to identify recurrences in the bladder early in their evolution in order to facilitate early intervention and ablation. Urine cytology is considered the gold standard urine biomarker. Although specificity exceeds 90% to 95%, its overall sensitivity ranges from 40% to 60% in expert hands and is both tumor grade and operator dependent. While cytology is an excellent test for detection of high grade disease, the sensitivity is particularly weak for the detection of low grade tumors. This has spawned an entire field of research of in vitro diagnostic tests and cell-based assays in order to improve the diagnostic accuracy for detection of incident or recurrent disease. To date, the US Food and Drug Administration approved dipstick and immunoassays marketed as point-of-care tests. The point-of-care tests are intended for use as an adjunct to cystoscopy and cytology, and may have a role in the office evaluation of hematuria patients. Monoclonal antibody-based tests combined with cytology may improve the diagnostic accuracy and are superior to cytology alone. A recently approved cell-based assay, utilizing fluorescent in situ hybridization technology, may help resolve suspicious cytologies, and provide early and additional information about the biology of the bladder urothelium beyond that provided by cytology, a marker of disease relatively late in evolution. Novel promising markers are in various stages of clinical testing, and a panel of biomarkers may serve in the future as a feasible alternative to urine cytology and cystoscopy for the screening, detection, and follow-up of non-muscle invasive bladder cancer. PMID- 15341677 TI - The epidermal growth factor receptor as a target for gastrointestinal cancer therapy. AB - The epidermal growth factor receptor (EGFR) is a member of the family of transmembrane protein kinase receptors known as the erbB or HER receptor family. When activated, EGFR phosphorylates and activates other intracellular proteins that affect cell signaling pathways, cellular proliferation, control of apoptosis and angiogenesis. EGFR signaling is best thought of as a network of activating and inactivating proteins with EGFR as the entry point into the network. EGFR overexpression occurs in most GI malignancies and while data are not entirely consistent, EGFR overexpression often confers a poor prognosis in those GI malignancies that have been studied. It often correlates with poorly differentiated histology, more advanced stage and other known poor prognostic markers. The EGFR is a tempting target because of its presence and overexpression on so many tumor types. However, downstream of the EGFR are several proteins that may be activated without EGFR thus allowing blockade to be overcome. Therefore, while blocking the activity of the EGFR protein appears to be a promising anticancer strategy, a simplistic strategy of blocking only EGFR is likely to only impact a minority of patients. It is time for the laboratory and clinical researchers to work closely together to develop this treatment strategy, moving back and forth from clinical to laboratory to best understand how to block this network effectively enough to produce a broader antitumor effect. While multiple methods of targeting the EGFR pathway are under development, including the inhibition of downstream proteins, only two modalities have entered clinical trials in GI malignancies: small molecule inhibitors of the intracellular kinase domain of EGFR and antibodies designed to block the extracellular ligand-binding domain of EGFR. EGFR inhibitors are still experimental in every GI malignancy with the notable exception of cetuximab that is approved for second or third-line therapy of metastatic colorectal cancer, used either alone or in combination with irinotecan (Camptosar, Kalamazoo, Mich). Data on clinical applications of these agents in GI malignancies will be the focus of this paper. PMID- 15341678 TI - Prevention and management of early esophageal cancer. AB - Early esophageal cancer is defined by its limitation to the esophageal mucosa and submucosa. It has become a curable malignant disease, in sharp contrast to the dismal prognosis of esophageal cancer at advanced stages, which still represents the majority of patients. Understanding the risk factors, establishing surveillance programs for patients at risk, and developing preventative interventions such as dietary and lifestyle changes or pharmacologic interventions hold the potential of reducing the incidence of the disease and of shifting the stage distribution toward early cancer. Endoscopic ultrasound examination is pivotal for distinguishing early from advanced stages of the disease because it allows for accurate assessment of tumor infiltration and regional lymph node involvement. The therapeutic mainstay for early esophageal cancer remains surgery. New, less invasive surgical techniques are being tested that are associated with less morbidity and mortality than standard radical esophagectomies. For patients who are not candidates for surgery, definitive chemoradiation is a viable alternative. New endoscopic ablation techniques, such as endoscopic mucosa resection and photodynamic therapy, are potential alternatives to surgery in patients with cancers limited to the mucosa. For patients with adenocarcinoma of the gastroesophageal junction with submucosal involvement, adjuvant chemoradiation should be considered because of its potential to increase survival. PMID- 15341679 TI - Strategies for prevention of colorectal cancer: pharmaceutical and nutritional interventions. AB - Interventions designed to delay or prevent the development of invasive colorectal adenocarcinoma might result in a profound impact on the incidence of and mortality from colorectal cancer. Current developmental efforts focus in two major directions: identification and testing of combination-targeted signal transduction pathway modulators and development of nutriceuticals. Both approaches recognize the heterogeneity of the carcinogenesis process. Combinations of agents (eg, aspirin or sulindac with DFMO, calcium with fiber), which have complementary or synergistic mechanisms or agents with broad spectrum anti-carcinogenic effects can potentially improve upon the effects of single agents and account for redundant signal transduction pathways involved in uncontrolled growth of clonal cells. Preliminary data in rodent systems suggest nutriceuticals or standardized foodstuffs, which contain multiple components, many of which are not identified have synergistic anticarcinogenesis effects. Diet modulation or supplementation with known standardized foodstuffs may be ideal in individuals at risk for colorectal cancer. Current clinical models for colorectal cancer prevention focus upon a pathologic-surrogate endpoint (reducing the recurrence or emergence of adenomas). This surrogate, generally accepted at a regulatory level, might not represent the true malignant progression from normal colonic mucosa to the malignant phenotype. Could the intervention simply prevent the emergence of adenomas that would not have progressed to invasive neoplasia in any event? Despite this major weakness, the adenoma remains the regulatory endpoint for efficacy because the cancer event remains rare, even among individuals selected as high-risk epidemiology but without known, highly pressured genetic-based stress. Over the next decade, the emergence of molecular and proteomic profiling tools have the potential of selecting adenomas that will progress to adenocarcinomas and will allow for investigation to be focused on individuals at increased risk. PMID- 15341680 TI - Regional therapy of liver metastases. AB - Liver metastases usually represent disseminated cancer; therefore, systemic chemotherapy is always appropriate. However, in some instances, the liver is the only site of metastasis and regional control of liver tumors may have important palliative and survival benefits, including a possibility of cure in selected patients. This review identifies unique patient subgroups that may benefit from regional therapy of liver metastases, and considers different treatment options and when these different options may be indicated. PMID- 15341681 TI - Arterial stiffness. PMID- 15341682 TI - Clinical trials report. Adverse prognostic significance of new diabetes in treated hypertensive subjects. PMID- 15341683 TI - Clinical trials report. Antihypertensive therapy for diabetic nephropathy. PMID- 15341684 TI - The use of selective aldosterone antagonists. AB - Mineralocorticoid hormones, specifically aldosterone, have been shown to be increasingly important in the development and maintenance of cardiovascular disorders, particularly hypertension and congestive heart failure. The use of the mineralocorticoid receptor blocker, spironolactone, has been fraught with side effects, largely related to the poor specificity of this agent. Eplerenone, a new and more specific mineralocorticoid receptor blocker with little effect on sex hormone receptors, has offered an alternative approach. Many studies in hypertension as well as a major and compelling study in congestive heart failure have documented the efficacy and specificity of eplerenone with a minimum of side effects. The major findings with this new agent are presented herein. PMID- 15341685 TI - Assessment of cardiovascular risk and choice of antihypertensive therapy. AB - Indications, choices of therapy, and goals of treatment for hypertension should be closely linked to the multivariate risk of cardiovascular events and ingredients of the vascular risk profile of patients with hypertension. Hypertension commonly occurs in combination with other cardiovascular disease risk factors, and the burden of the associated risk factors greatly affects the impact of elevated blood pressure and usually requires choices of specific antihypertensive agents shown to be particularly efficacious in the context of the associated risk factors or vascular conditions present. Guidelines assign highest priority to rigorous control of the blood pressure. Risk associated with hypertension has been shown to be incremental, with the level of blood pressure extending down to what was formerly considered the high-normal blood pressure range. In this "prehypertensive" state, drug treatment is justified only if the multivariate risk is substantial or there is target-organ damage. Most individuals with the metabolic syndrome, a condition commonly associated with hypertension, and singled out as an important consideration for treatment, are usually identified by the Framingham multivariate risk formulation. They require weight control and exercise, as well as specific choices of antihypertensive agents. PMID- 15341686 TI - Clinical importance of microalbuminuria in diabetes and hypertension. AB - Numerous studies document an almost linear association between the level of albuminuria and risk for a cardiovascular event. Recent data also demonstrate a strong association between the presence of microalbuminuria and elevations in C reactive protein. Therefore, the increased membrane permeability that generates microalbuminuria might be secondary to an inflammatory process. Progression from microalbuminuria to macroalbuminuria indicates worsening of vascular disease and presence of kidney disease. Recent pharmacologic interventions have resulted in significant delay and even arrest of progression of microalbuminuria to macroalbuminuria as well as kidney disease progression. Therefore, focus should be placed on use of antihypertensive agents that not only lower blood pressure but also lower or normalize albuminuria levels. All recent guideline statements support the use of angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs). Further lowering of albuminuria may be achieved by adding verapamil, diltiazem, or an ARB to an ACE inhibitor. PMID- 15341687 TI - Hypertension and dementia: does blood pressure control favorably affect cognition? AB - Dementia and aging are not synonymous. Dementia is a progressive deterioration in cognitive and social and/or occupational functions that can eventually impair a patient's ability to live independently. Alzheimer's disease is the most common form of dementia. It accounts for 50% to 70% of all patients with dementia. Vascular dementia, responsible for up to 15% of all diagnosed cases, is the second most common form of dementia. Hypertension remains a significant risk factor for vascular dementia. The optimal level of blood pressure control for the prevention of dementia and whether one particular class of antihypertensive drug is more beneficial than another remains uncertain. PMID- 15341689 TI - Clinical trials report. angiotensin-converting enzyme inhibition in diabetes: above and beyond the beneficial effect of blood pressure? PMID- 15341688 TI - Clinical trials report. Adverse prognostic significance of new diabetes in treated hypertensive subjects. PMID- 15341690 TI - Dual blockade of the renin angiotensin system in diabetic and nondiabetic kidney disease. AB - Dual blockade of the renin angiotensin system is based on a principle of obtaining the broadest and most efficient blockade of the effects of angiotensin II, by using the combination of an angiotensin-converting enzyme (ACE) inhibitor and an angiotensin II-receptor blocker (ARB). By combining two, different pharmacologic principles and inhibiting both the ACE and the angiotensin II type 1 receptor, it seems possible to block both the production and the action of angiotensin II, which would serve as efficient antihypertensive therapy. Exploring the beneficial effects of dual-blockade therapy is a work in rapid progress, in both diabetic and non-diabetic nephropathy. But evidence is also emerging in cardiovascular medicine, an overview of which is provided in this article. PMID- 15341691 TI - Remission and regression of diabetic nephropathy. AB - Diabetic kidney disease is considered to be an irreversible and inexorable progressive disease. Therefore, prevention of development of ESRD is extremely important. Animal studies have demonstrated that regression of existing renal morphologic lesions is feasible. In a sizable fraction of type 1 diabetic patients with overt nephropathy, remission (decrease in albuminuria to <300 mg/24 h) was obtained (31%), and regression of diabetic nephropathy (rate of decline in GFR < or = 1 mL/min/y), achieved (22%) by aggressive antihypertensive treatment. Furthermore, remission of nephrotic-range albuminuria (from >2500 mg/24 h to <600 mg/24 h) have been demonstrated to be achievable in one fifth of patients with nephrotic-range albuminuria. Remission of nephrotic-range albuminuria was associated with a reduction in decline in GFR and improved survival free of ESRD. However, to prevent development of ESRD in diabetic patients, an aggressive multifactorial approach, aiming at lowering blood pressure and albuminuria, and improving glycemic control, must be applied. PMID- 15341693 TI - Nocturnal hypertension: will control of nighttime blood pressure prevent progression of diabetic renal disease? AB - Patients with type 1 and 2 diabetes and nephropathy frequently have a blunted fall in nighttime arterial blood pressure. This abnormality is already seen in subjects with type 1 diabetes who are in the microalbuminuric phase of the disease, and we have also shown that an increase in nighttime systolic blood pressure precedes the development of microalbuminuria. These studies suggest that nocturnal hypertension may be an important early predictor of diabetic nephropathy. Various drugs have different effects on nocturnal blood pressure, and chronotherapy may be key in determining clinical outcomes. There is a compelling need for studies showing that treating nocturnal hypertension in diabetes can prevent renal disease progression. PMID- 15341692 TI - Antiproteinuric effect of RAS blockade: new mechanisms. AB - Experimental and clinical studies have shown that blockade of the renin angiotensin system (RAS) is effective in reducing proteinuria in conditions such as diabetes by reducing systemic and intraglomerular hydrostatic pressure. However, increasing evidence suggests that nonhemodynamic effects, such as preservation of the podocyte slit diaphragm structure and function, may also mediate the antiproteinuric effects of RAS blockade. In this review, we analyze in detail the evidence for known and novel mechanisms considered to play important roles in mediating the antiproteinuric effect of RAS blockers, with a particular focus on diabetic nephropathy. PMID- 15341695 TI - The dilemma of defining clinical severity in homozygous sickle cell disease. PMID- 15341694 TI - Blood pressure and progression of chronic kidney disease: importance of systolic, diastolic, or diurnal variation. AB - Several studies show that systolic blood pressure is an important predictor of renal disease progression, just as it is linked with cardiovascular consequences in hypertension. In contrast, particularly in older patients, diastolic blood pressure was not independently associated with risk of kidney disease progression in the same studies. Pulse pressure has been shown to be equivalent in predicting renal outcomes, but might not have added value after adjusting for systolic blood pressure. Several cross- sectional studies present a strong correlation of ambulatory blood pressure monitoring values with microalbuminuria, compared with office-based blood pressure measurements. Small, prospective studies have shown an association between loss of nocturnal blood pressure decline and outcomes, including microalbuminuria, accelerated kidney disease progression, and mortality. PMID- 15341696 TI - Comparison of oral versus intravenous phytonadione in patients with excessive anticoagulation. PMID- 15341697 TI - Splenectomy in immune thrombocytopenic purpura: recent controversies and long term outcomes. AB - Immune thrombocytopenic purpura (ITP) is an uncommon disorder but a relatively frequent cause of isolated thrombocytopenia. The pathogenesis of ITP involves enhanced platelet clearance, primarily in the spleen, caused by antiplatelet glycoprotein antibodies. Most patients with ITP respond transiently to an initial course of corticosteroids, although only 20% of adult patients achieve a durable remission after such therapy. Classically, patients who fail corticosteroid therapy undergo splenectomy, with approximately two thirds of these individuals maintaining partial remissions 5 years later. However, due to evidence suggesting a continuous relapse rate in splenectomized patients with ITP, in addition to the emergence of newer agents associated with durable remissions in ITP patients with intact spleens, the therapeutic approach to ITP is evolving. This article reviews the role of splenectomy in patients with ITP and reviews new approaches to management of ITP in patients who fail an initial trial of corticosteroids. PMID- 15341698 TI - Predicting hemorrhage using preoperative coagulation screening assays. AB - Each year in the United States, millions of units of fresh frozen plasma and platelets are transfused. A substantial proportion of these units are given to nonbleeding patients to "decrease the risk" of bleeding at the time of invasive procedures. The decision to transfuse is frequently triggered by an elevated prothrombin time (PT; international normalized ratio ) or activated partial thromboplastin time test result or by a depressed platelet count. This review summarizes published studies on the extent to which mild to moderate abnormalities of common laboratory tests of hemostasis reflect an increased risk of bleeding at the time of invasive bedside procedures, including central line placement, liver biopsy, paracentesis, thoracentesis, gastrointestinal endoscopy and biopsy, renal biopsy, bronchoscopy and transbronchial lung biopsy, and lumbar punctures. The poor predictive value of the PT (INR) is understandable in the context of the design and performance of this assay. Whether mild to moderate abnormalities of commonly used laboratory tests have any clinically relevant predictive value as appropriate triggers for prophylactic transfusions before invasive procedures should be investigated in formal randomized clinical trials. PMID- 15341699 TI - Update on the mechanism of action and future of activated prothrombin complex concentrates. AB - Activated prothrombin complex concentrates (aPCCs) are an established treatment for bleeding in patients with inhibitors. These products are derived from prothrombin complex concentrates, purified from human plasma with dedicated activation steps included in their manufacturing process. Despite these activation steps, the majority of the prothrombin complex proteins remain as zymogens, with only a relatively small content of activated coagulation enzymes. Among these, the order of concentration based on activity units is the following: factor VIIa to factor Xa to thrombin to factor IXa. Studies in various in vitro and in vivo model systems indicate that the mechanism of action of aPCCs is primarily based on an enzyme-substrate complex consisting of factor Xa and prothrombin. These findings are complemented by others showing that prothrombin is a major procoagulant that is capable of triggering hemostasis under physiologic and pathophysiologic conditions. Despite the findings of the mechanism of action, aPCCs have a long history of successful clinical use, with established dosing regimens, and a relatively low risk of thromboembolic complications compared with other treatment options for patients with inhibitors. PMID- 15341700 TI - All platelets are not equal: COAT platelets. AB - Collagen- and thrombin-activated (COAT) platelets were first described in 2000 and have attracted considerable interest, changing the interpretation of the way in which platelets contribute to thrombin generation and how their procoagulant activity is organized. Platelets activated by two agonists coming from glycoprotein VI or Fc gamma-receptor IIA agonists on the one hand and thrombin on the other produce a population of approximately 50% highly procoagulant active platelets. This subgroup is formed by tissue transglutaminase and factor XIIIa linking of serotonin to the procoagulant proteins from granules or plasma, and these serotonylated proteins bind to fibrinogen or thrombospondin on the platelet surface. Serotonylation in the platelet cytoplasm has recently been shown to be an important regulating mechanism governing the activation of small GTPases and their function in granule release. Recent studies with Tph-/- mice in which the peripheral serotonin, including that in platelets, is very strongly reduced, have shown a prolonged bleeding time, suggesting it has an important hemostatic role in the release of platelet von Willebrand factor. More knowledge about how COAT platelets are formed will be important for a better understanding of the physiology and pathology of hemostasis. PMID- 15341701 TI - Pathogenesis and management of bleeding in essential thrombocythemia and polycythemia vera. AB - Patients with essential thrombocythemia (ET) and polycythemia vera (PV) have a paradoxical predisposition to bleeding and thrombotic complications that are major causes of morbidity and mortality. Bleeding manifestations are often associated with extreme thrombocytosis that may lead to acquired von Willebrand syndrome (AVWS). Symptomatic AVWS, in this instance, is managed by platelet cytoreductive therapy and, in case of a life-threatening situation, platelet apheresis may be of additional value. Qualitative platelet defects are prevalent in PV and ET but have not been consistently linked to clinical bleeding. However, in vitro and in vivo hemostatic defects in these disorders are either precipitated or exacerbated by the use of aspirin or other nonsteroidal anti inflammatory drugs. Additional patient management issues are raised during systemic anticoagulation and surgery. This review summarizes putative pathogenetic mechanisms of bleeding and their management in ET and PV. PMID- 15341702 TI - Long-term, secondary treatment of deep venous thrombosis: do we know the appropriate duration of treatment or therapeutic regimen? PMID- 15341703 TI - New anticoagulants and their potential impact on the treatment of thromboembolic disease. AB - Traditional anticoagulants have drawbacks that make them complex to manage, limit their usefulness, and increase the possibility of adverse events. New anticoagulants are being developed that directly target a single coagulation factor. The agents have improved pharmacokinetics and pharmacodynamics and may not need coagulation monitoring. In addition, many are available orally. Agents that target factor Xa or factor IIa are the most advanced in development and of greatest interest. Fondaparinux and idraparinux are parenteral, specific, indirect, factor Xa inhibitors that have a mechanism of action similar to that of heparin. Idraparinux has a prolonged half-life and is dosed once weekly. Razaxaban is a small-molecule, oral, direct FXa inhibitor with demonstrated efficacy in orthopedic surgery for primary prevention of venous thromboembolism. Other oral Xa inhibitors are entering clinical trials. Ximelagatran is an oral factor IIa or thrombin inhibitor with documented efficacy for primary prevention of venous thromboembolism in orthopedic surgery, for the acute and chronic treatment of deep venous thrombosis and stroke prevention in atrial fibrillation. Other oral IIa inhibitors are entering clinical trials. Many of these agents will have a profound effect on the treatment of venous thromboembolism potentially resulting in reduced costs, improved patient satisfaction with treatment, and greater use of selected indications. PMID- 15341704 TI - Pharmacogenomics and the management of oral anticoagulation. AB - Warfarin, the principal oral anticoagulant used in the treatment and prevention of thromboembolic disease, is primarily metabolized by CYP2C9, one of the hepatic microsomal enzymes. Recent advances in the field of pharmacogenomics have identified the gene responsible for encoding CYP2C9, as well as several mutations that are associated with decreased warfarin metabolism. Clinically, patients with these mutations are more sensitive to the effects of warfarin, require lower warfarin dosing requirements to reach a particular therapeutic endpoint, and are at increased risk for over-anticoagulation and bleeding complications during both initiation and maintenance therapy. Routine pharmacogenomic screening has not yet been applied to clinical practice or analyzed for cost effectiveness but may be useful in identifying patients at risk for warfarin-related complications. PMID- 15341705 TI - Patient self-management of oral anticoagulation. AB - More than six million patients worldwide are being treated with an oral anticoagulant. Dosing of the anticoagulant is usually managed by a primary care physician, cardiologist, or a specialized anticoagulation clinic. With the introduction of point-of-care testing devices, it is possible for patients to monitor their coagulation time, the international normalized ratio, at home. Selected patients, when properly trained, are capable of managing their oral anticoagulation treatment from home, so-called patient self-management (PSM). PSM has been practiced for many years in Europe, where training centers and reimbursement are well established. Because the US Center for Medicare and Medicaid Services recently decided to cover PSM, the practice will likely become more common in the United States. This article reviews the evidence behind the efficacy, safety, and cost effectiveness of PSM. PMID- 15341706 TI - Extended treatment for venous thromboembolism: how long is long enough? AB - The long-term aim of treatment of venous thromboembolism (VTE) is the prevention of late recurrences and the post-thrombotic syndrome. The optimal duration of oral anticoagulant therapy after VTE is still controversial. Patients are currently stratified into risk categories for recurrence on the basis of clinical characteristics of the index VTE event. Patients with distal VTE or VTE caused by a transient risk factor are at low risk for recurrence and short-term anticoagulation (3 months) is indicated. Patients with an idiopathic event or with known thrombophilic defects such as factor V Leiden or the G20210A prothrombin mutation are candidates to a longer course of therapy (6 months). Patients with cancer, antiphospholipid antibodies syndrome, recurrent idiopathic event, antithrombin deficiency, protein C or protein S deficiency, homozygosity for factor V Leiden, or double heterozygosity are candidates for extended long term anticoagulation. More recently, studies have indicated that other factors such as D-dimer levels after the discontinuation of oral anticoagulant therapy or the residual vein thrombosis could be additional predictive factors for recurrences. PMID- 15341707 TI - Women's issues in venous thromboembolism. AB - Women and men experience venous thromboembolism (VTE) at similar rates, but there are several issues unique to women regarding this condition. For example, hormonal contraceptives and pregnancy increase the risk of VTE for women in their childbearing years, whereas hormone replacement therapy increases the risk for postmenopausal women. A woman's thrombotic risk must be considered when prescribing therapy for breast cancer because some treatments are associated with a higher incidence of VTE than others. In this paper, we review VTE in women, using cases from our clinical practice to illustrate some of the unique aspects of caring for this population. PMID- 15341708 TI - New endoscopic imaging techniques for improved detection of early neoplasia in patients with Barrett's esophagus. PMID- 15341710 TI - Intestinal epithelial defense systems protect against bacterial threats. AB - Numerous bacterial species inhabit the lumen of the human intestine. The epithelial cells that line the intestinal barrier are in direct contact with many of these species and have developed sophisticated strategies to prevent bacterial invasion of host tissue beyond simply providing a physical blockade. Intestinal epithelial cells (IECs) possess receptors that are capable of recognizing bacterial products, and engagement of these receptors results in the production and secretion of immunoregulatory proteins, such as cytokines and chemokines, which mobilize immune effector cells, including macrophages, dendritic cells, and neutrophils. In addition, IECs can produce various lipid-based eicosanoids that can contribute to the orchestration of the immune response. A better understanding of how the intestinal epithelium interacts with its microbial constituents may provide insight into strategies for treatment of enteric infections and inflammatory bowel disease. PMID- 15341711 TI - Evaluation and management of periampullary tumors. AB - Periampullary cancers make up 5% of all gastrointestinal cancers. The complexity of the periampullary anatomy makes determination of the origin of some of these tumors difficult. However, advances in imaging have helped with diagnosis as well as defining the extent of the lesion and its potential resectability. For many of these tumors, surgery is the recommended treatment. However, endoscopic removal is being extended to different lesions with encouraging preliminary results. Improvements in overall prognosis for periampullary tumors will be limited until diagnosis can be established earlier in the course of the disease and adjuvant therapies become more effective. PMID- 15341712 TI - Drug delivery to the small intestine. AB - Oral delivery of drugs to the small intestine is an important topic in the research and development of more effective oral dose forms. This review highlights several important developments in this area. An overriding theme in drug delivery to the small intestine is how to increase the efficiency (ie, how to increase bioavailability) of absorption. The role of P-glycoprotein and intestinal transporters is discussed in this regard. These systems are normally studied under defined in vitro conditions; recent data suggest that this approach, though useful, may not fully represent the in vivo situation. Recent advances and issues in the characterization and prediction of drug absorption from the small intestine are reviewed. These efforts, if successful, will shorten development timelines by eliminating compounds with poor absorption characteristics early in the process. Nanoparticulate delivery systems and those prepared by microfabrication technology are being used to improve bioavailability of poorly absorbed drugs. A relatively new technique (electroporation) has been proposed to enhance oral delivery of macromolecules, still an unrealized objective in drug delivery. PMID- 15341713 TI - Common variable immunodeficiency and the gastrointestinal tract. AB - Common variable immunodeficiency (CVID) is the second most prevalent primary immunodeficiency disorder but clinically the most important. It causes a wide spectrum of symptoms and signs affecting many systems of the body. CVID is a combination of humoral and cell-mediated deficiency, which explains not only why so many systems are affected but also why standard therapy in the form of intravenous immunoglobulin is not always effective. The gastrointestinal tract is the largest immune organ in the body, and it is therefore expected that this immunodeficiency will affect it in some way. The gastrointestinal manifestations of CVID are variable and tend to mimic known diseases, such as celiac sprue, pernicious anemia, and inflammatory bowel disease, but show significant differences on the microscopic level. Many studies continue to confirm a high prevalence of inflammatory, malignant, and infectious gastrointestinal disorders in patients with CVID. The T-cell-mediated defects of this immunodeficiency disorder are thought to be the cause of the majority of the gastrointestinal disorders in CVID and not the antibody deficiency. Therefore, intravenous immunoglobulin alone may be ineffective. Combination therapy with immunomodulators, such as azathioprine and 6-mercaptopurine, may be needed to treat these gastrointestinal manifestations of CVID. PMID- 15341714 TI - Electrifying progress for fecal incontinence. PMID- 15341715 TI - Magnetic resonance colonography: a promising new technique. AB - Colorectal carcinoma is still the second leading cause of cancer-related death, although it arises mostly from benign adenomas. Numerous screening methods are available, but none of them is accepted as ideal. Ultrafast three-dimensional data sets acquired by cross-sectional imaging modalities (CT or magnetic resonance ) in combination with new post-processing modes, known as virtual colonoscopy, have led to a new discussion of screening tests for colorectal cancer. Recently published results indicate a high sensitivity for CT colonography and for MR-based colonography (MRC), with detection rates of greater than 90% for colorectal lesions greater than 10 mm in size. Three-dimensional data acquisition for MRC is performed in less than 1 minute, and no severe complications have been reported. The main advantages of MRC are the lack of ionizing radiation, the low risk of the procedure, and low patient discomfort. MRC has become an attractive diagnostic procedure for colorectal lesions that can also be used as a screening method. PMID- 15341716 TI - Bowel preparation for gastrointestinal procedures. AB - Adequate colonic cleansing is essential for accurate and safe colonic procedures. Common preparations for cleansing include diet in combination with a cathartic agent, gut lavage, and phosphate preparations. The diet used with a cathartic consists of clear liquids or is designed to leave a minimal colonic fecal residue with laxatives. Gut lavage solutions wash out the colon in a safe and effective manner. Phosphate preparations offer an attractive alternative due to smaller volumes required for ingestion; however, electrolyte disturbances can occur. This review discusses the development and clinical experience with various colon cleansing regimens and efforts to improve the tolerability and safety of preparation for colonoscopy, virtual colonoscopy, colon surgery, barium enema, and flexible sigmoidoscopy. PMID- 15341717 TI - Pregnancy-related constipation. AB - Constipation is a common complaint in pregnancy. Its symptoms may include infrequent defecation, hard or scybalous stool, or excessive straining. An extensive evaluation is usually unnecessary for women who present with constipation for the first time during pregnancy. Most patients respond to dietary measures or simple laxatives. Few laxatives have been evaluated in clinical trials for use in pregnancy. Evidence supports treatment with fiber supplements and senna. The use of a pharmacologic agent for treatment of constipation during pregnancy must be weighed against possible adverse effects. Most laxatives carry a pregnancy category B or C classification. First-line therapy includes increasing fiber intake through diet or supplements. Osmotic laxatives may be beneficial for some patients. The short-term use of osmotic or stimulant laxatives is generally reserved for patients who fail to respond to dietary changes or bulking agents. PMID- 15341718 TI - Intestinal endometriosis: the great masquerader. AB - Chronic symptoms of abdominal pain and irregular bowel habits in women evoke a broad differential diagnosis including irritable bowel syndrome, infection, malabsorption, and inflammatory bowel disease. Endometriosis, a common disorder in young women that can involve the intestinal tract, deserves consideration as well. Intestinal endometriosis is typically asymptomatic; however, when symptoms occur, they can mimic those of irritable bowel syndrome. Identifying intestinal endometriosis can be challenging, but historical points and key clinical features aid in diagnosis. PMID- 15341719 TI - Acute colonic pseudoobstruction. AB - Acute colonic pseudoobstruction (ACPO) is a clinical condition of acute large bowel obstruction without mechanical blockage. ACPO occurs most often in hospitalized patients with serious underlying medical and surgical conditions. ACPO is an important cause of morbidity and mortality. The pathogenesis of ACPO is not completely understood but likely results from an imbalance in the autonomic regulation of colonic motor function. Metabolic or pharmacologic factors, as well as spinal or retroperitoneal trauma, may alter the autonomic regulation of colonic function, leading to excessive parasympathetic suppression or sympathetic stimulation. This imbalance results in colonic atony and pseudoobstruction. Early recognition and appropriate management are critical to minimizing morbidity and mortality. The mortality rate is estimated at 40% when ischemia or perforation occurs. The best documented treatment of ACPO is intravenous neostigmine, which leads to prompt decompression in the majority of patients after a single infusion. In patients failing or having contraindications to neostigmine, colonoscopic decompression is the active intervention of choice. Surgery is reserved for those with overt peritonitis or perforation. PMID- 15341720 TI - Recent advances in diverticular disease. AB - Diverticulosis is the most common structural abnormality of the colon. Although most people with colonic diverticula remain asymptomatic, a significant proportion of those affected develop symptoms and may suffer from complications. Whereas early research in diverticular disease addressed the pathogenesis of the condition, recent work has focused on symptomatic or complicated disease. Recent epidemiologic studies have attempted to identify patients who are at increased risk of perforated diverticular disease, and developing knowledge in neurogastroenterology has suggested mechanisms responsible for underlying chronically symptomatic disease. Further research in these areas has the potential to provide targeted therapeutic strategies for people with colonic diverticula. PMID- 15341721 TI - The Tudor tandem of 53BP1: a new structural motif involved in DNA and RG-rich peptide binding. AB - 53BP1 is a key transducer of the DNA damage checkpoint signal, which is required for phosphorylation of a subset of ATM substrates and p53 accumulation. After cell irradiation, the 53BP1 N-terminal region is phosphorylated. Its two C terminal BRCT motifs interact with p53. Its central region is required and sufficient for 53BP1 foci formation at DNA strand breaks and for 53BP1 binding to the kinetochore. It contains an RG-rich segment and interacts with DNA in vitro. Here we show that the major globular domain of the 53BP1 central region adopts a new structural motif composed of two tightly packed Tudor domains and a C terminal alpha helix. A unique surface essentially located on the first Tudor domain is involved in the binding to 53BP1 RG-rich sequence and to DNA, suggesting that the Tudor tandem can act as an adaptor mediating intramolecular as well as intermolecular protein-protein interactions and protein-nucleic acid associations. PMID- 15341722 TI - Determinants of functionality in the ubiquitin conjugating enzyme family. AB - The E2 enzymes are key enzymes in the ubiquitin and ubiquitin-like protein ligation pathways. To understand the functionality of the different E2 enzymes, we analyzed 190 protein sequences and 211 structures and electrostatic potentials. Key findings include: The ScUbc1 orthologs are defined by a C terminal UBA domain. An N-terminal sequence motif that is highly conserved in all E2s except for Cdc34 orthologs is important for the stabilization of the L7 loop and is likely to be involved in E1 binding. ScUbc11p has a different electrostatic potential from E2-Cp and other proteins with which it has high sequence similarity but different functionality. All the E2s known to ubiquitinate histones have a negative potential. The members of the NCUBE family have a positive electrostatic potential, although its form is different from that of the SUMO conjugating E2s. The specificities of only the ScUbc4/Ubc5 and ScUbc1p orthologs are reflected in their L4 and L7 loops. PMID- 15341723 TI - Crystallographic snapshots of Aspergillus fumigatus phytase, revealing its enzymatic dynamics. AB - Understanding of the atomic movements involved in an enzymatic reaction needs structural information on the active and inactive native enzyme molecules and on the enzyme-substrate, enzyme-intermediate, and enzyme-product(s) complexes. By using the X-ray crystallographic method, four crystal structures of Aspergillus fumigatus phytase were obtained at resolution higher than 1.7 A. The pH-dependent catalytic activity of A. fumigatus phytase was linked to three water molecules that may prevent the substrate from binding and thus block nucleophilic attack of the catalytic imidazole nitrogen. Comparison of various structures also identified the water molecule that attacks the phosphamide bond during the hydrolysis process, and established the hydrolysis pathway of the intermediate. Additionally, two reaction product phosphates were observed at the active site, suggesting a possible product release pathway after hydrolysis of the intermediate. These results can help explain the catalytic mechanism throughout the whole acid phosphatase family, as all key residues are conserved. PMID- 15341724 TI - Crystal structure of A. fulgidus Rio2 defines a new family of serine protein kinases. AB - The RIO family of atypical serine/threonine kinases contains two subfamilies, Rio1 and Rio2, highly conserved from archaea to man. Both RIO proteins from Saccharomyces cerevisiae catalyze serine phosphorylation in vitro, and the presence of conserved catalytic residues is required for cell viability. The activity of Rio2 is necessary for rRNA cleavage in 40S ribosomal subunit maturation. We solved the X-ray crystal structure of Archaeoglobus fulgidus Rio2, with and without bound nucleotides, at 2.0 A resolution. The C-terminal RIO domain is indeed structurally homologous to protein kinases, although it differs from known serine kinases in ATP binding and lacks the regions important for substrate binding. Unexpectedly, the N-terminal Rio2-specific domain contains a winged helix fold, seen primarily in DNA-binding proteins. These discoveries have implications in determining the target and function of RIO proteins and define a distinct new family of protein kinases. PMID- 15341725 TI - The crystal and solution structure of a putative transcriptional antiterminator from Mycobacterium tuberculosis. AB - We describe the crystal structure of Rv1626 from Mycobacterium tuberculosis at 1.48 A resolution and the corresponding solution structure determined from small angle X-ray scattering. The N-terminal domain shows structural homology to the receiver domains found in bacterial two-component systems. The C-terminal domain has high structural homology to a recently discovered RNA binding domain involved in transcriptional antitermination. The molecule in solution was found to be monomeric as it is in the crystal, but in solution it undergoes a conformational change that is triggered by changes in ionic strength. This is the first structure that links the phosphorylation cascade of the two-component systems with the antitermination event in the transcriptional machinery. Rv1626 belongs to a family of proteins, which we propose calling phosphorylation-dependent transcriptional antitermination regulators, so far only found in bacteria, and includes NasT, a protein from the assimilatory nitrate/nitrite reductase operon of Azetobacter vinelandii. PMID- 15341726 TI - Conformational changes of the flavivirus E glycoprotein. AB - Dengue virus, a member of the Flaviviridae family, has a surface composed of 180 copies each of the envelope (E) glycoprotein and the membrane (M) protein. The crystal structure of an N-terminal fragment of E has been determined and compared with a previously described structure. The primary difference between these structures is a 10 degrees rotation about a hinge relating the fusion domain DII to domains DI and DIII. These two rigid body components were used for independent fitting of E into the cryo-electron microscopy maps of both immature and mature dengue viruses. The fitted E structures in these two particles showed a difference of 27 degrees between the two components. Comparison of the E structure in its postfusion state with that in the immature and mature virions shows a rotation approximately around the same hinge. Flexibility of E is apparently a functional requirement for assembly and infection of flaviviruses. PMID- 15341727 TI - Novel catalytic mechanism of glycoside hydrolysis based on the structure of an NAD+/Mn2+ -dependent phospho-alpha-glucosidase from Bacillus subtilis. AB - GlvA, a 6-phospho-alpha-glucosidase from Bacillus subtilis, catalyzes the hydrolysis of maltose-6'-phosphate and belongs to glycoside hydrolase family GH4. GH4 enzymes are unique in their requirement for NAD(H) and a divalent metal for activity. We have determined the crystal structure of GlvA in complex with its ligands to 2.05 A resolution. Analyses of the active site architecture, in conjunction with mechanistic studies and precedent from the nucleotide diphosphate hexose dehydratases and other systems, suggest a novel mechanism of glycoside hydrolysis by GlvA that involves both the NAD(H) and the metal. PMID- 15341728 TI - Structure and RNA interactions of the N-terminal RRM domains of PTB. AB - The polypyrimidine tract binding protein (PTB) is an important regulator of alternative splicing that also affects mRNA localization, stabilization, polyadenylation, and translation. NMR structural analysis of the N-terminal half of PTB (residues 55-301) shows a canonical structure for RRM1 but reveals novel extensions to the beta strands and C terminus of RRM2 that significantly modify the beta sheet RNA binding surface. Although PTB contains four RNA recognition motifs (RRMs), it is widely held that only RRMs 3 and 4 are involved in RNA binding and that RRM2 mediates homodimerization. However, we show here not only that the RRMs 1 and 2 contribute substantially to RNA binding but also that full length PTB is monomeric, with an elongated structure determined by X-ray solution scattering that is consistent with a linear arrangement of the constituent RRMs. These new insights into the structure and RNA binding properties of PTB suggest revised models of its mechanism of action. PMID- 15341729 TI - The oxidized subunit B8 from human complex I adopts a thioredoxin fold. AB - Subunit B8 from ubiquinone oxidoreductase (complex I) (CI-B8) is one of several nuclear-encoded supernumerary subunits that are not present in bacterial complex I. Its solution structure shows a thioredoxin fold with highest similarities to the human thioredoxin mutant C73S and thioredoxin 2 from Anabeana sp. Interestingly, these proteins contain active sites in the same area, where the disulfide bond of oxidized CI-B8 is located. The redox potential of this disulfide bond is -251.6 mV, comparing well to that of disulfides in other thioredoxin-like proteins. Analysis of the structure reveals a surface area that is exclusively composed of highly conserved residues and thus most likely a subunit interaction site within complex I. PMID- 15341730 TI - Novel binding site identified in a hybrid between cholera toxin and heat-labile enterotoxin: 1.9 A crystal structure reveals the details. AB - A hybrid between the B subunits of cholera toxin and Escherichia coli heat-labile enterotoxin has been described, which exhibits a novel binding specificity to blood group A and B type 2 determinants. In the present investigation, we have determined the crystal structure of this protein hybrid, termed LCTBK, in complex with the blood group A pentasaccharide GalNAcalpha3(Fucalpha2)Galbeta4(Fucalpha3)GlcNAcbeta, confirming not only the novel binding specificity but also a distinct new oligosaccharide binding site. Binding studies revealed that the new specificity can be ascribed to a single mutation (S4N) introduced into the sequence of Escherichia coli heat-labile enterotoxin. At a resolution of 1.9 A, the new binding site is resolved in excellent detail. Main features include a complex network of water molecules, which is well preserved by the parent toxins, and an unexpectedly modest contribution to binding by the critical residue Asn4, which interacts with the ligand only via a single water molecule. PMID- 15341731 TI - Crystal structures of the type III effector protein AvrPphF and its chaperone reveal residues required for plant pathogenesis. AB - The avrPphF locus from Pseudomonas syringae pv. phaseolicola, the causative agent of bean halo-blight disease, encodes proteins which either enhance virulence on susceptible hosts or elicit defense responses on hosts carrying the R1 resistance gene. Here we present the crystal structures of the two proteins from the avrPphF operon. The structure of AvrPphF ORF1 is strikingly reminiscent of type III chaperones from bacterial pathogens of animals, indicating structural conservation of these specialized chaperones, despite high sequence divergence. The AvrPphF ORF2 effector adopts a novel "mushroom"-like structure containing "head" and "stalk" subdomains. The head subdomain possesses limited structural homology to the catalytic domain of bacterial ADP-ribosyltransferases (ADP-RTs), though no ADP-RT activity was detected for AvrPphF ORF2 in standard assays. Nonetheless, this structural similarity identified two clusters of conserved surface-exposed residues important for both virulence mediated by AvrPphF ORF2 and recognition of this effector by bean plants expressing the R1 resistance gene. PMID- 15341732 TI - Crystal structure of the carboxyltransferase domain of acetyl-coenzyme A carboxylase in complex with CP-640186. AB - Acetyl-coenzyme A carboxylases (ACCs) are important targets for the development of therapeutic agents against obesity, diabetes, and other diseases. CP-640186 is a potent inhibitor of mammalian ACCs and can reduce body weight and improve insulin sensitivity in test animals. It is believed to target the carboxyltransferase (CT) domain of these enzymes. Here we report the crystal structure of the yeast CT domain in complex with CP-640186. The inhibitor is bound in the active site at the interface of a dimer of the CT domain. CP-640186 has tight interactions with the putative biotin binding site in the CT domain and demonstrates a distinct mode of inhibiting the CT activity as compared to the herbicides that inhibit plant ACCs. The affinity of inhibitors for the CT domain has been assessed using kinetic and fluorescence anisotropy binding studies. The structural information identifies three regions for drug binding in the active site of CT. PMID- 15341733 TI - Solution structure of human initiation factor eIF2alpha reveals homology to the elongation factor eEF1B. AB - The GTP-bound form of the trimeric eukaryotic translation initiation factor 2 (eIF2) transfers aminoacylated initiator methionyl tRNA onto the 40S ribosome. We have solved with solution NMR the structure of the alpha subunit of human eIF2 (heIF2alpha). The protein consists of two domains that are mobile relative to each other. The N-terminal domain has an S1-type oligonucleotide/oligosaccharide binding-fold subdomain and an alpha-helical subdomain. The C-terminal domain adopts an alphabeta-fold very similar to the C-terminal domain of elongation factor (eEF) 1Balpha, the guanine-nucleotide exchange factor for eEF1A. The structural and functional similarities found between eIF2alpha/eIF2gamma and eEF1Balpha/eEF1A suggest a model for the interaction of eIF2alpha with eIF2gamma, and eIF2 with Met-tRNAiMet. It further indicates a previously unrecognized evolutionary lineage of eIF2alpha/gamma from the functionally related elongation factor eEF1Balpha/eEF1A complex. PMID- 15341734 TI - Crystal structure of 7,8-dihydropteroate synthase from Bacillus anthracis: mechanism and novel inhibitor design. AB - Dihydropterate synthase (DHPS) is the target for the sulfonamide class of antibiotics, but increasing resistance has encouraged the development of new therapeutic agents against this enzyme. One approach is to identify molecules that occupy the pterin binding pocket which is distinct from the pABA binding pocket that binds sulfonamides. Toward this goal, we present five crystal structures of DHPS from Bacillus anthracis, a well-documented bioterrorism agent. Three DHPS structures are already known, but our B. anthracis structures provide new insights into the enzyme mechanism. We show how an arginine side chain mimics the pterin ring in binding within the pterin binding pocket. The structures of two substrate analog complexes and the first structure of a DHPS-product complex offer new insights into the catalytic mechanism and the architecture of the pABA binding pocket. Finally, as an initial step in the development of pterin-based inhibitors, we present the structure of DHPS complexed with 5-nitro-6-methylamino isocytosine. PMID- 15341735 TI - The CAP-Gly domain of CYLD associates with the proline-rich sequence in NEMO/IKKgamma. AB - CYLD was originally identified as the human familial cylindromatosis tumor suppressor. Recently, it was reported that CYLD directly interacts with NEMO/IKKgamma and TRAF2 in the NF-kappaB signaling pathway. The two proteins bind to a region of CYLD that contains a Cys-box motif and the third cytoskeleton associated protein-glycine conserved (CAP-Gly) domain. Here we report that the third CAP-Gly domain of CYLD specifically interacts with one of the two proline rich sequences of NEMO/IKKgamma. The tertiary structure of the CAP-Gly domain shares the five-stranded beta sheet topology with the SH3 domain, which is well known as a proline-rich sequence-recognition domain. However, chemical shift mapping revealed that the peptide binding site of the CAP-Gly domain is formed without the long peptide binding loop characteristic of the SH3 domain. Therefore, CAP-Gly is likely to be a novel proline-rich sequence binding domain with a mechanism different from that of the SH3 domain. PMID- 15341736 TI - Structure of superoxide reductase bound to ferrocyanide and active site expansion upon X-ray-induced photo-reduction. AB - Some sulfate-reducing and microaerophilic bacteria rely on the enzyme superoxide reductase (SOR) to eliminate the toxic superoxide anion radical (O2*-). SOR catalyses the one-electron reduction of O2*- to hydrogen peroxide at a nonheme ferrous iron center. The structures of Desulfoarculus baarsii SOR (mutant E47A) alone and in complex with ferrocyanide were solved to 1.15 and 1.7 A resolution, respectively. The latter structure, the first ever reported of a complex between ferrocyanide and a protein, reveals that this organo-metallic compound entirely plugs the SOR active site, coordinating the active iron through a bent cyano bridge. The subtle structural differences between the mixed-valence and the fully reduced SOR-ferrocyanide adducts were investigated by taking advantage of the photoelectrons induced by X-rays. The results reveal that photo-reduction from Fe(III) to Fe(II) of the iron center, a very rapid process under a powerful synchrotron beam, induces an expansion of the SOR active site. PMID- 15341737 TI - An asymmetric complex of restriction endonuclease MspI on its palindromic DNA recognition site. AB - Most well-known restriction endonucleases recognize palindromic DNA sequences and are classified as Type IIP. Due to the recognition and cleavage symmetry, Type IIP enzymes are usually found to act as homodimers in forming 2-fold symmetric enzyme-DNA complexes. Here we report an asymmetric complex of the Type IIP restriction enzyme MspI in complex with its cognate recognition sequence. Unlike any other Type IIP enzyme reported to date, an MspI monomer and not a dimer binds to a palindromic DNA sequence. The enzyme makes specific contacts with all 4 base pairs in the recognition sequence, by six direct and five water-mediated hydrogen bonds and numerous van der Waal contacts. This MspI-DNA structure represents the first example of asymmetric recognition of a palindromic DNA sequence by two different structural motifs in one polypeptide. A few possible pathways are discussed for MspI to cut both strands of DNA, either as a monomer or dimer. PMID- 15341738 TI - Efficiency of information transmission by retinal ganglion cells. AB - BACKGROUND: Different types of retinal ganglion cells convey different messages to the brain. Messages are in the form of spike patterns, and the number of possible patterns per second sets the coding capacity. We asked if different ganglion cell types make equally efficient use of their coding capacity or whether efficiency depends on the message conveyed. RESULTS: We recorded spike trains from retinal ganglion cells in an in vitro preparation of the guinea pig retina. By calculating, for the observed spike rate, the number of possible spike patterns per second, we calculated coding capacity, and by counting the actual number of patterns, we estimated information rate. Cells with "brisk" responses, i.e., high firing rates, and a general message transmitted information at high rates (21 +/- 9 bits s(-1)). Cells with "sluggish" responses, i.e., lower firing rates, and specific messages (direction of motion, local-edge) transmitted information at lower rates (13 +/- 7 bits s(-1)). Yet, for every type of ganglion cell examined, the information rate was about one-third of coding capacity. For every ganglion cell, information rate was very close (within 4%) to that predicted from Poisson noise and the cell's actual time-modulated rate. CONCLUSIONS: Different messages are transmitted with similar efficiency. Efficiency is limited by temporal correlations, but correlations may be essential to improve decoding in the presence of irreducible noise. PMID- 15341739 TI - Positive selection on MMP3 regulation has shaped heart disease risk. AB - BACKGROUND: The evolutionary forces of mutation, natural selection, and genetic drift shape the pattern of phenotypic variation in nature, but the roles of these forces in defining the distributions of particular traits have been hard to disentangle. To better understand the mechanisms contributing to common variation in humans, we investigated the evolutionary history of a functional polymorphism in the upstream regulatory region of the MMP3 gene. This single base pair insertion/deletion variant, which results in a run of either 5 or 6 thymidines 1608 bp from the transcription start site, alters transcription factor binding and influences levels of MMP3 mRNA and protein. The polymorphism contributes to variation in arterial traits and to the risk of coronary heart disease and its progression. RESULTS: Phylogenetic and population genetic analysis of primate sequences indicate that the binding site region is rapidly evolving and has been a hot spot for mutation for tens of millions of years. We also find evidence for the action of positive selection, beginning approximately 24,000 years ago, increasing the frequency of the high-expression allele in Europe but not elsewhere. Positive selection is evident in statistical tests of differentiation among populations and haplotype diversity within populations. Europeans have greater arterial elasticity and suffer dramatically fewer coronary heart disease events than they would have had this selection not occurred. CONCLUSIONS: Locally elevated mutation rates and strong positive selection on a cis-regulatory variant have shaped contemporary phenotypic variation and public health. PMID- 15341740 TI - SKAR is a specific target of S6 kinase 1 in cell growth control. AB - BACKGROUND: The mammalian target of rapamycin (mTOR) and phosphatidylinositol 3 kinase (PI3K) signaling pathways promote cell growth and cell cycle progression in response to nutritional, energy, and mitogenic cues. In mammalian cells, the ribosomal protein S6 kinases, S6K1 and S6K2, lie downstream of mTOR and PI3K, suggesting that translational control through the phosphorylation of S6 regulates cell growth. Interestingly, genetic experiments predict that a substrate that is specific to S6K1 but not S6K2 regulates cell growth. RESULTS: Here we identify SKAR as a novel and specific binding partner and substrate of S6K1 but not S6K2. We find that serines 383 and 385 of human SKAR are insulin-stimulated and rapamycin-sensitive S6K1 phosphorylation sites. Quantitative mass spectrometry reveals that serine 383/385 phosphorylation is sensitive to RNA interference (RNAi)-mediated S6K1 reduction, but not S6K2 reduction. Furthermore, RNAi mediated reduction of SKAR decreases cell size. SKAR is nuclear protein with homology to the Aly/REF family of RNA binding proteins, which has been proposed to couple transcription with pre-mRNA splicing and mRNA export. CONCLUSIONS: We have identified a novel and specific target of S6K1, SKAR, which regulates cell growth. The homology of SKAR to the Aly/REF family links S6K1 with mRNA biogenesis in the control of cell growth. PMID- 15341741 TI - Smad affinity can direct distinct readouts of the embryonic extracellular Dpp gradient in Drosophila. AB - BACKGROUND: The TGF-beta signaling molecule Decapentaplegic (Dpp) is an essential morphogen that patterns many tissues during Drosophila development, including the embryonic dorsal ectoderm and larval wing imaginal disk. An activity gradient of Dpp specifies distinct cell fates in the dorsal ectoderm of the embryo through the activation of different transcriptional threshold responses. RESULTS: We have analyzed the gene Race, which is expressed in response to peak levels of Dpp signaling in gastrulating embryos. We show that the Smad transcription factors, which are intracellular transducers of Dpp signaling, are essential activators of Race in vivo. Furthermore, increasing the affinity of the Smad binding sites in the Race enhancer broadens the expression pattern of a linked reporter gene and alters its behavior in mutant embryos to that characteristic of a distinct threshold response. CONCLUSIONS: Smad activator affinity is a critical determinant of the threshold response to the extracellular Dpp gradient in the embryo. Our results identify a mechanism for interpreting the Dpp gradient in the embryo which is different to the reciprocal repressor gradient model proposed for the wing disk. We suggest that transcription factor binding site affinity will be a general strategy used in the interpretation of other extracellular morphogen gradients. PMID- 15341742 TI - Visually induced plasticity of auditory spatial perception in macaques. AB - When experiencing spatially disparate visual and auditory stimuli, a common percept is that the sound originates from the location of the visual stimulus, an illusion known as the ventriloquism effect. This illusion can persist for tens of minutes, a phenomenon termed the ventriloquism aftereffect. The underlying neuronal mechanisms of this rapidly induced plasticity remain unclear; indeed, it remains untested whether similar multimodal interactions occur in other species. We therefore tested whether macaque monkeys experience the ventriloquism aftereffect similar to the way humans do. The ability of two monkeys to determine which side of the midline a sound was presented from was tested before and after a period of 20-60 min in which the monkeys experienced either spatially identical or spatially disparate auditory and visual stimuli. In agreement with human studies, the monkeys did experience a shift in their auditory spatial perception in the direction of the spatially disparate visual stimulus, and the aftereffect did not transfer across sounds that differed in frequency by two octaves. These results show that macaque monkeys experience the ventriloquism aftereffect similar to the way humans do in all tested respects, indicating that these multimodal interactions are a basic phenomenon of the central nervous system. PMID- 15341743 TI - Archer fish learn to compensate for complex optical distortions to determine the absolute size of their aerial prey. AB - Many animals, including humans, can visually judge the absolute size of objects regardless of changes in viewing distance and thus despite the resulting dramatic differences in the size of the actual retinal images. For animals that have to judge the size of aerial objects from underwater views, this can be a formidable problem; our calculations show that considerable and strongly viewpoint-dependent corrections are needed to compensate for the effects of light refraction. Archer fish face these optical difficulties because they have to shoot down aerial insects over a wide range of horizontal and vertical distances. We show here that these fish can learn to acquire size constancy with remarkable precision and are thus fully capable of taking complex viewpoint dependency into account. Moreover, we demonstrate that archer fish solve the problem not by interpolating within a set of stored views and distances but by learning the laws that connect apparent size with the fish's relative position to the target. This enables the fish to readily judge the absolute sizes of objects from completely novel views. PMID- 15341744 TI - Feo, the Drosophila homolog of PRC1, is required for central-spindle formation and cytokinesis. AB - We performed a functional analysis of fascetto (feo), a Drosophila gene that encodes a protein homologous to the Ase1p/PRC1/MAP65 conserved family of microtubule-associated proteins (MAPs). These MAPs are enriched at the spindle midzone in yeast and mammals and at the fragmoplast in plants, and are essential for the organization and function of these microtubule arrays. Here we show that the Feo protein is specifically enriched at the central-spindle midzone and that its depletion either by mutation or by RNAi results in aberrant central spindles. In Feo-depleted cells, late anaphases showed normal overlap of the antiparallel MTs at the cell equator, but telophases displayed thin MT bundles of uniform width instead of robust hourglass-shaped central spindles. These thin central spindles exhibited diffuse localizations of both the Pav and Asp proteins, suggesting that these spindles comprise improperly oriented MTs. Feo-depleted cells also displayed defects in the contractile apparatus that correlated with those in the central spindle; late anaphase cells formed regular contractile structures, but these structures did not constrict during telophase, leading to failures in cytokinesis. The phenotype of Feo-depleted telophases suggests that Feo interacts with the plus ends of central spindle MTs so as to maintain their precise interdigitation during anaphase-telophase MT elongation and antiparallel sliding. PMID- 15341745 TI - Contrast, probability, and saccadic latency; evidence for independence of detection and decision. AB - Many factors influence how long it takes to respond to a visual stimulus. The lowest-level factors, such as luminance and contrast, determine how easily different elements of a target can be detected. Higher-level factors are to do with whether these elements constitute a stimulus requiring a response; they include prior probability and urgency. It is natural to think of these two processes, detection and decision, as occurring in series, so that overall reaction time is essentially the sum of the contributions of each stage. Here, measurements of saccadic latency to visual targets whose contrast and prior probability are systematically manipulated demonstrate that there are indeed separable stages of detection and decision. Both can be quantitatively described by rise-to-threshold mechanisms; the average rate of rise of the first is a simple logarithmic function of target contrast, whereas the second shows the linear rise characteristic of the LATER model of neural decision making. The implication is that under normal, high-contrast conditions, in which detection is very fast, the random variability that is characteristic of all reaction times is not caused by sensory noise but is gratuitously introduced by the brain itself; paradoxically, by conferring unpredictability it may aid an organism's survival. PMID- 15341746 TI - Mos mediates the mitotic activation of p42 MAPK in Xenopus egg extracts. AB - The ERK1/ERK2 MAP kinases (MAPKs) are transiently activated during mitosis, and MAPK activation has been implicated in the spindle assembly checkpoint and in establishing the timing of an unperturbed mitosis. The MAPK activator MEK1 is required for mitotic activation of p42 MAPK in Xenopus egg extracts; however, the identity of the kinase that activates MEK1 is unknown. Here we have partially purified a Cdc2-cyclin B-induced MEK-activating protein kinase from mitotic Xenopus egg extracts and identified it as the Mos protooncoprotein, a MAP kinase kinase kinase present at low levels in mitotic egg extracts, early embryos, and somatic cells. Immunodepletion of Mos from interphase egg extracts was found to abolish Delta90 cyclin B-Cdc2-stimulated p42 MAPK activation. In contrast, immunodepletion of Raf-1 and B-Raf, two other MEK-activating kinases present in Xenopus egg extracts, had little effect on cyclin-stimulated p42 MAPK activation. Immunodepletion of Mos also abolished the transient activation of p42 MAPK in cycling egg extracts. Taken together, these data demonstrate that Mos is responsible for the mitotic activation of the p42 MAPK pathway in Xenopus egg extracts. PMID- 15341747 TI - EFF-1 is sufficient to initiate and execute tissue-specific cell fusion in C. elegans. AB - Despite the identification of essential processes in which cell fusion plays spectacular roles such as in fertilization and development of muscle, bone, and placenta, there are no identified proteins that directly mediate developmental cell fusion reactions. C. elegans has recently become among the best characterized models to use for studying developmental cell fusion. The eff-1 (epithelial fusion failure) gene encodes novel type I membrane proteins required for epithelial cell fusion. Analysis of eff-1 mutants showed that cell fusion normally restricts routes for cell migration and establishes body and organ shape and size [ 5, 8, 9, 11]. Here, we explored cell fusion by using time-lapse confocal and electron microscopy of different organs. We found that ectopic expression of eff-1 is sufficient to fuse epithelial cells that do not normally fuse. This ectopic fusion results in cytoplasmic content mixing and disappearance of apical junctions, starting less than 50 min after the start of eff-1 transcription. We found that eff-1 is necessary to initiate and expand multiple microfusion events between pharyngeal muscle cells. Surprisingly, eff-1 is not required to fuse the gonadal anchor cell to uterine cells. Thus, eff-1 is sufficient and essential for most but not all cell fusion events during C. elegans development. PMID- 15341748 TI - Vocal-tract filtering by lingual articulation in a parrot. AB - Human speech and bird vocalization are complex communicative behaviors with notable similarities in development and underlying mechanisms. However, there is an important difference between humans and birds in the way vocal complexity is generally produced. Human speech originates from independent modulatory actions of a sound source, e.g., the vibrating vocal folds, and an acoustic filter, formed by the resonances of the vocal tract (formants). Modulation in bird vocalization, in contrast, is thought to originate predominantly from the sound source, whereas the role of the resonance filter is only subsidiary in emphasizing the complex time-frequency patterns of the source (e.g., but see ). However, it has been suggested that, analogous to human speech production, tongue movements observed in parrot vocalizations modulate formant characteristics independently from the vocal source. As yet, direct evidence of such a causal relationship is lacking. In five Monk parakeets, Myiopsitta monachus, we replaced the vocal source, the syrinx, with a small speaker that generated a broad-band sound, and we measured the effects of tongue placement on the sound emitted from the beak. The results show that tongue movements cause significant frequency changes in two formants and cause amplitude changes in all four formants present between 0.5 and 10 kHz. We suggest that lingual articulation may thus in part explain the well-known ability of parrots to mimic human speech, and, even more intriguingly, may also underlie a speech-like formant system in natural parrot vocalizations. PMID- 15341749 TI - Scc2 couples replication licensing to sister chromatid cohesion in Xenopus egg extracts. AB - The cohesin complex is a central player in sister chromatid cohesion, a process that ensures the faithful segregation of chromosomes in mitosis and meiosis. Previous genetic studies in yeast show that Scc2/Mis4, a HEAT-repeat-containing protein, is required for the loading of cohesin onto chromatin. In this study, we have identified two isoforms of Scc2 in humans and Xenopus (termed Scc2A and Scc2B), which are encoded by a single gene but have different carboxyl termini created by alternative splicing. Both Scc2A and Scc2B bind to chromatin concomitant with cohesin during DNA replication in Xenopus egg extracts. Simultaneous immunodepletion of Scc2A and Scc2B from the extracts impairs the association of cohesin with chromatin, leading to severe defects in sister chromatid pairing in the subsequent mitosis. The loading of Scc2 onto chromatin is inhibited in extracts treated with geminin but not with p21(CIP1), suggesting that this step depends on replication licensing but not on the initiation of DNA replication. Upon mitotic entry, Scc2 is removed from chromatin through a mechanism that requires cdc2 but not aurora B or polo-like kinase. Our results suggest that vertebrate Scc2 couples replication licensing to sister chromatid cohesion by facilitating the loading of cohesin onto chromatin. PMID- 15341751 TI - Pushing the boundaries. PMID- 15341750 TI - Dynamics of yeast Myosin I: evidence for a possible role in scission of endocytic vesicles. AB - Cortical actin patches are dynamic structures required for endocytosis in yeast. Recent studies have shown that components of cortical patches localize to the plasma membrane in a precisely orchestrated manner, and their movements at and away from the plasma membrane may define the endocytic membrane invagination and vesicle scission events, respectively. Here, through live-cell imaging, we analyze the dynamics of the highly conserved class I unconventional myosin, Myo5, which also localizes to cortical patches and is known to be involved in endocytosis and actin nucleation. Myo5 exhibits a pattern of dynamic localization different from all cortical patch components analyzed to date. Myo5 associates with cortical patches only transiently and remains stationary during its brief cortical lifespan. The peak of Myo5 association with cortical patches immediately precedes the fast movement of Arp2/3 complex-associated structures away from the plasma membrane, thus correlating precisely with the proposed vesicle scission event. To further test the role of Myo5, we generated a temperature-sensitive mutant myo5 allele. In the myo5 mutant cells, Myo5 exhibits a significantly extended cortical lifespan as a result of a general impairment of Myo5 function, and Arp2 patches exhibit an extended slow-movement phase prior to the fast movement toward the cell interior. The myo5 mutant cells are defective in fluid phase endocytosis and exhibit an increased number of invaginations on the membrane. Based on these results, we hypothesize that the myosin I motor protein facilitates the membrane fusion/vesicle scission event of endocytosis. PMID- 15341752 TI - Q & A. PMID- 15341753 TI - Back to the future. PMID- 15341754 TI - Mammalian fertilization. PMID- 15341755 TI - The real 'kingdoms' of eukaryotes. PMID- 15341757 TI - Tumor outbreaks in marine turtles are not due to recent herpesvirus mutations. PMID- 15341758 TI - Natural selection: sign, sign, everywhere a sign. AB - Natural selection is an important factor influencing variation in the human genome, but most genetic studies of natural selection have focused on variants with unknown phenotypic associations. This trend is changing. New studies are rapidly revealing the effects of natural selection on genetic variants of known or likely functional importance. PMID- 15341759 TI - Microtubule dynamics: faint speckle, hidden dragon. AB - The results of recent experiments in budding and fission yeast show that there is a diversity of mechanisms for targeting proteins to the plus ends of microtubules in eukaryotic cells. PMID- 15341760 TI - Development: dissecting the dynamics of segment determination. AB - A novel combination of high-resolution time-course expression data and computational modelling has provided a remarkably detailed picture of a key stage of Drosophila segment determination, highlighting the dynamic nature of this process. PMID- 15341761 TI - MAPK signaling: Sho business. AB - Sho1 is a membrane protein in yeast that activates the Hog MAPK signaling pathway in response to high osmolarity. An accumulating body of work has focused on Sho1 as a model to better understand the mechanisms that dictate signaling specificity. PMID- 15341762 TI - Membrane targeting: glued by a lipid to the ER. AB - Opi1p, a transcription regulator of phospholipid metabolism in budding yeast, is retained in the endoplasmic reticulum by association with Scs2p. New research shows that binding of phosphatidic acid to Opi1p is a prerequisite for this targeting. PMID- 15341763 TI - Limb movement: getting a handle on grasp. AB - The mechanical complexity of the hand is indisputable, but there is increasing evidence that its control is simplified in many tasks around synergic groups of muscles that effectively decrease the number of controlled degrees of freedom. PMID- 15341764 TI - Animal development: crowd control. AB - To shape a developing animal, individual cell movements must be coordinated over long distances. Two recent studies help show how this is achieved during convergence and extension of the Drosophila germ-band, where polarity within the plane of the embryonic epithelium biases junction remodeling to polarize cell intercalation. PMID- 15341765 TI - Tumor metastasis: a new twist on epithelial-mesenchymal transitions. AB - Epithelial-mesenchymal transitions are essential for normal embryonic development and for progression of non-invasive tumor cells into malignant, metastatic carcinomas. Twist, an important regulator of morphogenesis in the embryo, has recently been implicated in the onset of invasive behavior during tumor progression. PMID- 15341766 TI - SIN and the art of splitting the fission yeast cell. AB - The septation initiation network (SIN) triggers the onset of cytokinesis in the fission yeast Schizosaccharomyces pombe by promoting contraction of the medially placed F-actin ring. SIN signaling is regulated by the polo-like kinase plo1p and by cdc2p, the initiator of mitosis, and its activation is co-ordinated with other events in mitosis to ensure that cytokinesis does not begin until chromosomes have been separated. Though the SIN controls the contractile ring, the signal originates from the poles of the mitotic spindle. Recent studies suggest that the spindle pole body may act as a dynamic assembly site for active SIN signaling complexes. In the budding yeast Saccharomyces cerevisiae the counterpart of the SIN, called the MEN, mediates both mitotic exit and cytokinesis, in part through regulating activation of the phosphoprotein phosphatase Cdc14p. Flp1p, the S. pombe ortholog of Cdc14p, is not essential for mitotic exit, but may contribute to an orderly mitosis-G1 transition by regulating the destruction of the mitotic inducer cdc25p. PMID- 15341767 TI - Indel in the FIC1/ATP8B1 gene-a novel rare type of mutation associated with benign recurrent intrahepatic cholestasis. AB - Benign recurrent intrahepatic cholestasis (BRIC) is a rare inherited liver disease characterized by recurrent attacks of severe cholestasis with no progression to end stage liver disease. Patients have jaundice, however, serum gamma-glutamyltransferase and cholesterol levels remain within the normal range during the attacks. Three mutations in the familial intrahepatic cholestasis 1 (ATP8B1) gene encoding a P-type ATPase have been reported so far in patients with the autosomal recessive form of BRIC. A novel rare type insertion-deletion mutation, also called indel, was found in exon 24 of ATP8B1 in our patient together with a known missense mutation 1982T>C in exon 17. The mechanism of the indel formation is proposed and impact of the indel mutation on the function of ATP8B1 protein is discussed. PMID- 15341768 TI - Rebound hepatitis following withdrawal of immunosuppressive therapy in patients with chronic Hepatitis B viral infections. AB - Background: Rebound hepatitis is a potentially life-threatening complication of withdrawal from immunosuppressive therapy in patients with chronic Hepatitis B viral (HBV) infections. Objectives: To document the incidence of rebound hepatitis and determine whether the hepatitis is associated with serologic evidence of immunological rebound or the appearance of specific mutations in the HBV genome. Methods: Serum cytokines (IL-6, IL-10, TNF-alpha and INF-gamma) were documented by enzyme linked immunoassays and previously described HBV mutants (surface, core, pre-core and basal core promoter) by signal probe hybridization analysis in chronic HBV carriers treated with either 6 weeks of prednisone followed by 6 weeks of acyclovir (PR/AC, n = 20) or placebo/placebo (PL/PL, n = 20). Results: Rebound hepatitis (serum ALT > 2X baseline) occurred in 6/20 (30%) PR/AC patients versus 2/20 (10%) PL/PL recipients (P = 0.24). Serum cytokine levels were similar in those who developed rebound hepatitis compared to those who did not. HBV mutants were absent prior to and during treatment but developed in the follow-up period in three patients. All three patients were PR/AC recipients and in each case, the HBV mutation was in the basal core promoter gene. In two of the three patients, the mutant appeared just prior to the onset of rebound hepatitis while in the third, rebound hepatitis did not occur. Conclusions: The results of this study indicate an association exists between some cases of rebound hepatitis and the development of HBV mutants. PMID- 15341769 TI - The fluctuations of viral load and serum alanine aminotransferase levels in chronic hepatitis C. AB - Although viral load of hepatitis C virus (HCV) is a predictor of response to interferon therapy, little is known about its fluctuations. We assessed its fluctuations and their correlation with serum alanine aminotransferase (ALT) levels. Viral load was prospectively measured bimonthly for 22 months in 109 patients. In 40 patients, viral load changed more than five fold. Changes were transient and always returned to the baseline levels. ALT levels changed more than three fold in 30 patients. Changes of viral load accompanied simultaneous changes of ALT levels in only 7 of 40 patients with changes of viral load. Mean viral load in 22 months was significantly correlated with mean ALT levels inversely (r = 0.278, P = 0.0036). Mean viral load was significantly higher in 27 patients with persistently normal ALT levels (452.0 +/- 342.5pg/ml) than in 30 patients with changes of ALT levels (202.4 +/- 215.0pg/ml) (P = 0.0016) and than in 52 patients without changes of ALT levels (301.1 +/- 295.4pg/ml) (P = 0.0458). Inverse correlation of viral load with ALT levels suggests that viral load is in suppression by inflammatory activity. However, changes of ALT levels infrequently accompanied simultaneous changes of viral load and vice versa, as often seen in chronic hepatitis B virus infection. PMID- 15341770 TI - Brain glutamine and glutamate levels in patients with liver cirrhosis: assessed by 3.0-T MRS. AB - Background/Objectives: Magnetic resonance spectroscopic (MRS) studies have revealed abnormal metabolism in the brain of patients with liver cirrhosis, including an increase in total brain glutamine (Gln) and glutamate (Glu) levels (Glx). However, with conventional MRS techniques, it was difficult to separate the Glx signals. Using a high-magnetic field MR equipment and a newly developed data processing method, we attempted to separate the Glx signals on an MRS. Subjects and Methods: Twenty-three patients with liver cirrhosis and 11 healthy adults were enrolled in this study. After designating a region of interest in the occipital lobe gray matter of each subject, [Formula: see text] (proton)-MRS was performed using 3.0-T MR equipment. Results: MRS conducted using the 3.0-T MR equipment allowed Gln signals in the brain to be distinguished from the Glx signals. The brain signal intensity of Gln was found to be significantly higher in the liver cirrhosis group (0.658 +/- 0.23) than in the control group (0.473 +/ 0.08) (P < 0.05). Neither the Glu nor the Gln signal intensity showed any correlation with the blood ammonia level. Conclusion: High-magnetic field MRS allowed us to separate the Glx signals in the brain and revealed that the increase in the total brain Glu and Gln levels in patients is solely attributable to an increase in the level of Gln. PMID- 15341771 TI - Detection of serum and intrahepatic KL-6 in anti-HCV positive patients with hepatocellular carcinoma. AB - We investigated the clinical significance of serum and intrahepatic KL-6/MUC1 (KL 6) in patients with hepatitis C virus (HCV) antibody-positive hepatocellular carcinoma (HCC). The subjects included 76 patients diagnosed with anti-HCV positive HCC, 69 with, and 51 without, liver cirrhosis (LC). Frozen serum samples were obtained from each subject to determine the serum KL-6 levels using an enzyme-linked immunosorbent assay. Expression of KL-6 antigen in the liver was also investigated using immunoperoxidase staining. The mean serum KL-6 level in patients with HCC was [Formula: see text] U/ml (319U/ml for HCC with LC, 342.8U/ml for HCC without LC). Serum KL-6 levels in patients with HCC with LC and HCC without LC did not differ. Serum KL-6 levels were elevated with increases in the size of spaces occupied by tumors in the liver. Among patients with HCC, there was no correlation between serum KL-6 levels and alpha-fetoprotein (AFP) levels and protein induced by vitamin K absence or antagonist-II (PIVKA-II) levels. However, some patients with low levels of AFP and PIVKA-II possessed high levels of KL-6. Furthermore, serum KL-6 levels decreased after therapy for HCC nodules. Immunohistochemical staining showed KL-6 antigen was detected within the cell membrane and in the cytoplasm of cancer cells. KL-6 antigen was localized on the membrane and the endoplasmic reticulum of cancer cells in the cancerous foci by electron microscopy. Our results suggest that serum KL-6 levels represent a serological marker of HCC development, because KL-6 expression was localized to the cancer cells in HCC nodules. PMID- 15341772 TI - Transforming growth factor-beta-induced stimulation of formation of collagen fiber network and anti-fibrotic effect of taurine in an in vitro model of hepatic fibrosis. AB - The cell strain M, which was established from normal rat liver cells, is characterized by the active formation of a collagen fiber network. In this study, we investigated the characterization of M cells and evaluated the anti-fibrogenic effects of taurine using this culture system. M cells expressed cytokeratin (CK)8, CK18, vimentin, and alpha-smooth muscle actin, whereas expression of CK-19 or desmin was not detected. Also, M cells expressed transforming growth factor (TGF)-beta1, -beta2, and TGF-beta type I and II receptors, and treatment with TGF beta1 (1ng/ml) for 6 days markedly stimulated the formation of a collagen fiber network and expression of procollagen alpha1(I) mRNA. When M cells were treated with various concentrations of taurine (10-50mM), network formation and procollagen alpha1(I) expression were significantly suppressed in a dose dependent manner. Additionally, even in the presence of TGF-beta1, taurine treatment effectively reduced the formation of a collagen fiber network. These results suggest that M cells exhibit features of not only hepatocytes but also myofibroblasts, and TGF-beta1 plays an important role in the formation of collagen fiber networks in this culture system. Additionally, this M cell culture system is appropriate for use as an in vitro model of hepatic fibrosis in the evaluation of the anti-fibrogenic effects of various agents. PMID- 15341773 TI - Hepatic infarction following selective hepatic artery embolization with microcoils for iatrogenic biliary hemorrhage. AB - Purpose: The aim of this study was to explore the relation of collateral filling to ischemic or infarcted liver following selective embolization of hepatic artery with microcoils in patients with iatrogenic hemobilia. Methods: We performed retrospective analysis of clinical outcomes and post-embolization angiograms in eight patients (mean age of 66 years) studied over the last 7 years. Hemobilia occurred after percutaneous biliary drainage (n = 5) and percutaneous hepatic biopsy (n = 3). Causes of bleeding were pseudoaneurysm (n = 6), arterial laceration (n = 1), and direct hepatic artery-to-biliary duct fistula (n = 1). We placed microcoils in the subsegmental (n = 4) or segmental branch (n = 2), or both branches (n = 2), distal and proximal to the bleeding point. Results: We obtained complete hemostasis in all patients (100%). Four patients had no hepatic infarction after embolization. Normal filling of the distal part of the embolized branch through collaterals was seen on post-embolization films. Four patients with no collateral filling experienced liver infarction in the area corresponding to embolized branch. One patient with severe portal stenosis died of hepatic failure. Conclusion: Hepatic infarction is related to lack of immediate collateral flow. PMID- 15341774 TI - A case of merged idiopathic portal hypertension in course of mixed connective tissue disease. AB - We report a case of idiopathic portal hypertension (IPH) complicated with mixed connective tissue disease (MCTD). The patient, a 41-year-old woman, was admitted to another hospital because of tarry stools in March 2000. Emergency endoscopy revealed bleeding from the esophageal varices, and she was referred to our hospital. Twelve years before, she had been diagnosed as having MCTD, because she exhibited Raynaud's phenomenon, pleuritis, pericarditis, and had high titers of anti nuclear antibody (ANA) and anti-U1 ribonucleoprotein (RNP). Laboratory examinations, imaging examinations, and liver biopsy indicated that the esophageal varices were caused by IPH. The association of IPH and MCTD is very rare; to the best of our knowledge, only five cases of MCTD, associated with pulmonary hypertension (PH), have been reported. PMID- 15341775 TI - Peptibodies: the new cool technology. PMID- 15341776 TI - Reactome: clear view of a starry sky. PMID- 15341777 TI - NIH chemical genomics network takes off. PMID- 15341778 TI - Arnold O. Beckman, inventor and philanthropist. PMID- 15341779 TI - An integrated, multidisciplinary approach for drug safety assessment. PMID- 15341780 TI - The quiet revolution: outsourcing in pharma. PMID- 15341781 TI - JAK kinase inhibitors. PMID- 15341782 TI - Mastering medicinal chemistry: strategies, issues and success stories. PMID- 15341783 TI - The transdermal revolution. AB - Historically, developments in transdermal drug delivery have been incremental, focusing on overcoming problems associated with the barrier properties of the skin, reducing skin irritation rates and improving the aesthetics associated with passive patch systems. More-recent advances have concentrated on the development of non-passive systems to aid delivery of larger drug molecules, such as proteins and nucleotides, as the trend for discovering and designing biopharmaceuticals continues. Fundamentally, improvements in transdermal delivery will remain incremental until there is wider acceptance of this route of administration within the pharmaceutical industry. Only then will the transdermal revolution live up to its true potential. PMID- 15341784 TI - The HIF pathway as a therapeutic target. AB - Hypoxia-inducible factor (HIF) is an alpha,beta-heterodimeric transcription factor that mediates cellular responses to low oxygen concentration via the transcriptional activation of specific genes involved in both tumorogenesis and angiogenesis. Manipulation of the HIF pathway has potential use for the treatment of ischemic disease and cancer. Unlike HIF-beta, which is constitutively expressed, the levels and activity of the HIF-alpha subunit are regulated by processes involving posttranslational hydroxylation, catalyzed by Fe(II)- and 2 oxoglutarate-dependent oxygenases. This review focuses on the HIF pathway as a therapeutic target. PMID- 15341785 TI - Transporter-mediated drug delivery: recent progress and experimental approaches. AB - A comprehensive list of drug transporters has recently become available as a result of extensive genome analysis, as well as membrane physiology and molecular biology studies. This review covers recent progress in identification and characterization of drug transporters, illustrative cases of successful drug delivery to, or exclusion from, target organs via transporters, and novel experimental approaches to therapeutics using heterologously transduced transporters in tissues. We aim to provide clues that could lead to efficient strategies for the use of transporters to deliver drugs and/or to optimize lead compounds. PMID- 15341790 TI - Intra-medial prefrontal administration of SCH-23390 attenuates ERK phosphorylation and long-term memory for trace fear conditioning in rats. AB - The prefrontal cortex is known to be involved in the acquisition of trace conditioning, a higher-cognitive form of Pavlovian conditioning in which a conditioned stimulus and an unconditioned stimulus are separated by a time gap. We have recently reported that medial prefrontal (mPFC) extracellular-signal regulated kinase (Erk) phosphorylation is involved in the long-term memory storage of trace fear conditioning. Because of the important role dopamine D1 receptors play in prefrontal function, such as working memory, and due to evidence that dopamine D1 receptor activity can modulate plasticity, we investigated their role in prefrontal Erk phosphorylation following trace fear conditioning. We found that inhibition of dopamine D1 receptors through intra mPFC infusion of SCH-23390 (1 microg/0.5 microL) 15 min prior to trace fear conditioning resulted in a decrease in training-related Erk phosphorylation. Additionally, pre-training intra-mPFC infusion of SCH-23390 also resulted in the impairment of long-term retention of CS-US association. These findings implicate mPFC dopamine D1 receptor activity in the storage of long-term memory for higher cognitive associative tasks. PMID- 15341791 TI - Trace fear conditioning is reduced in the aging rat. AB - Auditory trace fear conditioning is a hippocampus-dependent learning task that requires animals to associate an auditory conditioned stimulus (CS) and a fear producing shock-unconditioned stimulus (US) that are separated by an empty 20-s trace interval. Previous studies have shown that aging impairs learning performance on hippocampus-dependent tasks. This study measured heart rate (HR) and freezing fear responses to determine if aging impairs hippocampus-dependent auditory trace fear conditioning in freely moving rats. Aging and Young rats received one long-trace fear conditioning session (10 trials). Each trial consisted of a tone-CS (5 s) and a shock-US separated by an empty 20-s trace interval. The next day rats received CS-alone retention trials. Young rats showed significantly larger HR and freezing responses on the initial CS-alone retention trials compared to the Aging rats. A control group of aging rats received fear conditioning trials with a short 1-s trace interval separating the CS and US. The Aging Short-Trace Group showed HR and freezing responses on the initial CS alone retention trials that were similar to the Young Long-Trace Group, but greater than the Aging Long-Trace Group. A second aging control group received unpaired CSs and USs, and showed no HR or freezing responses on CS-alone retention trials. These data show that HR and freezing are effective measures for detecting aging related deficits in trace fear conditioning. PMID- 15341792 TI - Time of day-dependent latent inhibition of conditioned taste aversions in rats. AB - We have determined that the temporal context of drinking can modulate latent inhibition of learned saline aversions in Wistar rats by changing the time of day of drinking of the preexposure and conditioning phases. Latent inhibition was absent in the group preexposed and conditioned to saline at different times of the day, but not in the group that was preexposed and conditioned at the same time of day. The results confirm a previous report that the time of day can modulate taste aversion learning independently of other environmental cues. It is proposed that the features and duration of the habituation training to the temporal contexts used may be critical for time-dependent latent inhibition to appear. PMID- 15341793 TI - The influence of NMDA receptors in the dorsomedial striatum on response reversal learning. AB - In mammals, the dorsomedial striatum is one brain area shown to be critical for the flexible shifting of response patterns. At present, the neurochemical mechanisms that underlie learning during a shift in response patterns are unknown. The present study examined the effects of NMDA competitive antagonist, DL-2-amino-5-phosphonopentanoic acid (AP-5), injected into the dorsomedial striatum on the acquisition and reversal of a response discrimination. Male Long Evans rats were tested across two consecutive days in a modified cross-maze. Rats received an infusion of either saline or AP-5 (5 or 25 nmol) 5 min prior to each test session. In the acquisition phase rats learned to turn in one direction (right or left) to receive a cereal reinforcement. In the reversal learning phase rats learned to turn in the opposite direction as in the acquisition phase. In both phases, criterion was achieved when a rat made 10 consecutive correct trials. Infusions of AP-5 did not impair acquisition, but impaired reversal learning of a response discrimination in a dose-dependent fashion. The reversal learning deficit induced by AP-5 resulted from reversions back to the originally learned response pattern following the initial shift. These results suggest that activation of NMDA receptors in the dorsomedial striatum are critical for the flexible shifting of response patterns by enhancing the reliable execution of a new response pattern under changing task contingencies. PMID- 15341794 TI - Effects of pre- or post-training paradoxical sleep deprivation on two animal models of learning and memory in mice. AB - The aim of the present study was to verify the effects of pre- or post-training paradoxical sleep (PS) deprivation in mice tested in the passive and the plus maze discriminative avoidance tasks. Three-month-old Swiss male mice were placed in narrow platforms in a water tank for 72 h to prevent the occurrence of PS. Control animals were kept in the same room, but in their home cages. Before or after this period, the animals were submitted to the training session of one of the behavioral tasks. The test sessions were performed 3 and 10 days after the training. The animals that were PS-deprived before the training session showed retention deficits in the test sessions performed 3 days later in both tasks (decreased latency to enter the dark chamber of the passive avoidance apparatus or increased percent time spent in the aversive arm of the plus-maze discriminative avoidance apparatus). Animals that were PS deprived after the training session showed no differences from control animals in the test sessions performed 3 days after the training in any of the tasks, but showed passive and discriminative avoidance retention deficits in the test performed 10 days after the training. The results suggest that both pre- and post-training paradoxical sleep deprivation produce memory deficits in mice. However, these effects have different temporal characteristics. PMID- 15341795 TI - Context-specific habituation of the zenk gene response to song in adult zebra finches. AB - Zebra finches show a multifaceted response to playback of tape-recorded birdsong. A novel song induces an overt listening behavior, accompanied by a distinct electrophysiological signature and a wave of gene expression in the auditory telencephalon. With repetition of the same song these responses all habituate; habituation can persist for a day or more and is specific for the repeated song. This habituation is a form of selective memory. Here we describe four experiments to probe the nature of the information stored in that memory, using habituation of the genomic response as a physiological assay for stimulus recognition. Adult male zebra finches (awake and unrestrained) were exposed to repetitions of a test song to habituate the genomic response. The same song was then presented in one of four different contexts: (1) from a speaker on the opposite side of the cage; (2) at a reduced sound pressure level; (3) paired with constant illumination of colored lights next to the speaker; (4) paired with colored lights that were turned on and off in synchrony with each song bout. In all cases except the third, a "familiar" (habituated) song re-induced a new wave of gene expression in the auditory telencephalon when presented in the new context. These results reveal that memory for a specific song, as indicated by the initial gene habituation, incorporates more than just an acoustic description of the song. We suggest that habituation in the auditory telencephalon is controlled in part by an extrinsic system that allows detection of synchronous activity in different sensory or representational modalities. PMID- 15341796 TI - The effects of hippocampal lesions on learning, memory, and reward expectancies. AB - The hippocampus appears to be critical for the formation of certain types of memories. Hippocampal-lesioned animals fail to exhibit some spatial, contextual, and relational associations. After aspiration lesions of the hippocampus and/or cortex, male rats were allowed to recover for three weeks before being trained on a matching-to-position task. The matching-to-position task was altered to influence the type of cognitive strategies a subject would use to solve the task. The main behavioral manipulation was the reinforcement contingency assignment: Use of a differential outcomes procedure (DOP) or a nondifferential outcomes procedure (NOP). The DOP involves correlating each to-be-remembered event with a distinct reward condition via Pavlovian trace conditioning, whereas the NOP results in random reward contingency. We found that hippocampal lesions did retard learning the matching rule, regardless of the reinforcement contingency assignment. However, when delay intervals were added to the task memory performance of subjects with hippocampal lesions was dramatically impaired--if subjects were not trained with the DOP. When subjects were trained with the DOP, the hippocampal lesion had a marginal effect on delayed memory performance. These findings demonstrate two important points regarding lesions of the hippocampus: (1) hippocampal lesions have a minimal effect on the on the ability of rats to use reward information to solve a delayed discrimination task; (2) rats with hippocampal lesions have the ability to learn about reward information using Pavlovian trace conditioning procedures. PMID- 15341797 TI - Bilateral lesion of the cerebellar-dentate nucleus impairs egocentric sequential learning but not egocentric navigation in the rat. AB - The involvement of the cerebellum in procedural learning is demonstrated in visuomotor-sequence tasks, as lesion of this area impedes the acquisition of new sequences. Likewise, the lateral cerebellum appears to be involved in the acquisition of new sequences, but not in the execution of learned sequences. In contrast, the dentate nucleus participates only in the execution of learned visuomotor sequences. In previous studies, disruption of the procedural elements of spatial navigation following cerebellar or dentate lesions has been reported. However, as praxic strategies (egocentric learning) are included in the procedural elements of the navigation, the participation of the cerebellar dentate nucleus in egocentric procedural learning processes has not been evaluated. Therefore, using colchicine, bilateral lesions were made in the cerebellar-dentate nucleus of Sprague-Dawley rats, and these rats were given two tasks: egocentric-based motor sequence learning in the radial maze and egocentric navigation in the Morris water maze. The lesioned rats were unable to use the sequential information in the short term and showed delayed long-term acquisition, which was probably due to the inability to detect the sequence. No effects on the egocentric navigation task were observed. Our results indicate that the cerebellar-dentate nucleus is involved in the detection of egocentric sequential information but not in the use of this information in the navigation process. Further, they show differential involvement of the cerebellar-dentate nucleus in the execution of learned visuomotor sequences, as the dentate lesion disrupted the acquisition of new egocentric-motor-based sequences. PMID- 15341798 TI - Extinction-induced immobility in the water maze and its neurochemical concomitants in aged and adult rats: a possible model for depression? AB - Extinction of escape behavior in the water maze due to the removal of the platform, was hypothesized to induce a negative state, including the development of immobility, which is held to reflect a state of "despair" when measured in the forced swimming test. 27 aged and 8 adult animals (26 and 3 months old Wistar rats, respectively) were tested in the water maze during nine days with a platform hidden, followed by 7 days of extinction trials with the platform absent. As expected, both age groups developed immobility over the extinction trials, with the aged showing more than the adults. To examine whether the age difference in immobility was related to performance differences during acquisition, the aged were subdivided into superior, intermediate and inferior learners (n = 9 per group) on the basis of overall times to platform during acquisition, and compared with each other and the adults. Results showed that the aged inferior learners displayed the highest levels of immobility among the aged. Immobility scores were then correlated with post-mortem neurotransmitter contents in the hippocampus and ventral striatum. In the ventral striatum, levels of immobility were correlated with levels of acetylcholine, dopamine and the metabolite dihydroxyphenylacetic acid in the aged, and with norepinephrine in the adults. The data support the hypothesis that multiple extinction trials in the water maze result in immobility that may indicate "behavioral despair," and that striatal neurotransmitter systems correlate with the degree of its expression. The concept of extinction-induced despair is held to provide the promise of a conceptual and empirical model of human depression that is the consequence of loss of reinforcers. PMID- 15341799 TI - Estrogen replacement in ovariectomized rats affects strategy selection in the Morris water maze. AB - While estrogen enhances performance on some tasks of learning and memory, it has impairing or no effects on others. It has been proposed that estrogen differentially affects performance on various tasks of learning and memory by influencing the strategy used to solve a task. The goal of the present study was to determine if estrogen would influence strategy selection in the Morris water maze. Long-Evans rats were ovariectomized and implanted with Silastic capsules containing 25% estradiol diluted in cholesterol or 100% cholesterol. Rats were trained in a water maze task in which multiple strategies were available for use to locate a hidden escape platform that was moved to a new location for each set of four daily trials. During 10 days of acquisition trials, a visible floating landmark was always located in a static position relative to the hidden escape platform. Additionally, fixed extramaze cues visible to the animals surrounded the maze. Following acquisition, 2 days of probe trials were conducted in which the static landmark was removed. Estrogen replacement in ovariectomized rats resulted in impaired performance across 10 days of acquisition. Additionally, while removal of the visible landmark during the probe trials had no effect on the performance of the females receiving estrogen, it significantly disrupted performance of females receiving cholesterol treatment. These results indicate that estrogen replacement in ovariectomized rats biases an animal against using a landmark or static cue to aid in the location of a hidden escape platform in the water maze. PMID- 15341800 TI - Is the unilateral lesion of the left substantia nigra pars compacta sufficient to induce working memory impairment in rats? AB - Adult male Wistar rats with a substantia nigra pars compacta (SNc) lesion induced by intranigral administration of 1 micromol 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) were used as a model of early phase Parkinson's disease (PD). This lesion caused a partial depletion of striatal dopamine (DA). The animals were submitted to a spatial working memory version of the water maze task in which they had to find a hidden (submersed) platform using online-maintained information that the platform remains in the same place during four consecutive trials, but that it is moved to another place every training day. Left, but not right SNc-lesioned rats were impaired in finding the platform in the second trial. This result suggests that the left SNc plays a key role in spatial working memory. Control experiments ruled out the possibility that motor impairment, sensory neglect, and/or impairment in the mental representation of the contralateral spatial environment had affected performance of the SNc-lesioned rats. PMID- 15341801 TI - Inducible repression of CREB function disrupts amygdala-dependent memory. AB - Evidence from Aplysia, Drosophila, mice, and rats indicates that the CREB (cAMP/Ca2+ responsive element binding protein) family of transcription factors is critical for long-term memory. Recent findings, however, suggest that performance abnormalities may contribute to the memory deficits attributed to CREB manipulations in mammals. To clarify the role of CREB in memory, we used a paradigm, conditioned taste avoidance, that places few performance demands on the subject. We show that lesioning or blocking protein synthesis in the basolateral amygdala of mice disrupts conditioned taste aversion. Furthermore, either chronically or acutely disrupting CREB function in two different types of genetically modified mice blocks memory for conditioned taste aversion measured 24 h following training. Together, these findings indicate that CREB-mediated transcription and protein synthesis are required for conditioned taste aversion memory. PMID- 15341802 TI - Conditioned taste aversion with sucrose and tactile stimuli in the pond snail Lymnaea stagnalis. AB - A new form of taste aversion conditioning was established in the pond snail Lymnaea stagnalis. An associative memory, lasting 24h, was produced in the pond snail with 20 pairings of 100 mM sucrose as the conditioned stimulus (CS) and mechanical stimulation to the head as the unconditioned stimulus (UCS). Animals exposed to reverse pairings of the CS and UCS failed to learn the association. The learning was characterized by a shift in the response to the UCS from a whole body withdrawal response to the cessation of feeding behavior. PMID- 15341803 TI - Urinary incontinence: historical, global, and epidemiologic perspectives. AB - Urinary incontinence (UI) is a devastating worldwide problem in older adults. Establishing the true prevalence and incidence of UI remains a challenge as different societies define and approach UI differently. Nevertheless, most societies share the common practice of stigmatizing elders with incontinence. Enhanced awareness of global perspectives and epidemiological trends of incontinence isa necessary pre-requisite to improving management of this distressing geriatric syndrome. PMID- 15341804 TI - Physiology of incontinence. AB - The physiology of incontinence is related to the normal physiologic mechanisms of aging and to abnormal pathologic changes that recently have become better understood. Further research is needed to develop new methods of pharmacologic treatment. PMID- 15341805 TI - Urinary incontinence and the community-dwelling elder: a practical approach to diagnosis and management for the primary care geriatrician. AB - Although there have been dramatic improvements in the management of incontinence since the 1940s, this area remains a major challenge to the geriatrician. Patients need to be asked regularly if they are incontinent and, after the diagnosis is made, should be given a rational treatment plan. Urodynamics should not be performed until mainstream treatments have failed. PMID- 15341806 TI - Urinary incontinence in the nursing home: practical approach to evaluation and management. AB - This article provides a practical, evidence-based approach to the evaluation and treatment of the older incontinent nursing home resident. The concept of targeting treatment to those individuals who have cognitive and sensorimotor impairments and who are most likely to benefit is discussed. The efficacy and feasibility of comprehensive continence care programs, the use of clinical care guidelines in the nursing home, and the need for translational research in this area are addressed briefly. PMID- 15341807 TI - Urinary incontinence in the frail elder. AB - Urinary incontinence (UI) is a complex disorder affecting a large segment of the frail older population. Physical factors such as medication use and disease comorbidity can affect continence control. Cognitive disorders and functional decline pose additional challenges in management of urinary symptoms. Those individuals who have adequate socioeconomic support and caregiver assistance may have a greater advantage in maintaining continence.Physical, cognitive, functional, and psychosocial factors each contribute to the risk profile for both the development of frailty and the likelihood of experiencing UI. PMID- 15341808 TI - Urinary incontinence and the terminally ill older person. AB - The principles of managing urinary incontinence in an older patient who has a terminal illness should be based on the general principles of overall care for terminally ill older persons. First, health care professionals need to understand "where" the person is in the dying process. Second, they must be able to predict, with as much accuracy as possible, the consequences of any action or inaction(that is, a decision made not to do an intervention that typically is done). Third, they must understand how the patient's symptom is uncomfortable and bothersome from the patient's standpoint. All three steps need to take into account the family's perception of the patient's discomfort, and, whenever possible, the family should be involved in the decision making. PMID- 15341809 TI - Urodynamic evaluation of the older adult: bench to bedside. AB - Urinary symptoms are prevalent in the geriatric population.Symptoms, however, often are misleading. Urodynamic evaluation can help target specific treatments of specific disorders. In this article, the fundamentals of urodynamics are described. In addition,urodynamic findings in common geriatric conditions, such as Parkinson's disease, cerebral vascular accidents, benign prostatic hypertrophy, and incontinence, are discussed. PMID- 15341810 TI - Caring for the incontinent elder: advanced practice nursing concepts. AB - Urinary incontinence (UI) in the older adult is a multisystem health problem that has an impact on the lives of millions of older adults. Advanced practice nurses (APNs) specializing in gerontology,family, and adult practice are equipped educationally with the expertise to prevent, assess, and manage this health problem. The emerging emphasis on UI prevention will help guide the practice of APNs in the future. APNs will serve older adults well in prevention and management of UI by capitalizing on their roles as expert clinicians, researchers, consultants, educators, and collaborators with the interdisciplinary teams. PMID- 15341811 TI - Urinary incontinence: behavioral modification therapy in older adult. AB - Urinary incontinence is common in the elderly population;however, less than half the people who have this problem are examined or treated. The Agency for Health Care Policy and Research recommends behavioral interventions before other forms of treatment in patients who are diagnosed with stress, urge, or mixed incontinence after a basic evaluation. Behavioral treatments are safe, effective, and low risk and have no documented side effects. They also can be used as an adjunct to other therapies, such as pharmacologic and surgical interventions. PMID- 15341812 TI - Pharmacologic management of urinary incontinence. AB - Urinary incontinence is a highly prevalent and difficult to treat medical condition. For those who are affected, urinary incontinence produces profound modifications in lifestyle and perceived quality of life. Pharmacologic therapy clearly is beneficial for most types of urinary incontinence. The low therapeutic success of current treatments and obnoxious side effects, however, limit patient satisfaction and compliance. There is a considerable need for improved therapeutic options to treat this devastating condition. PMID- 15341813 TI - Surgical management of urinary incontinence: a geriatric perspective. AB - Urinary incontinence (UI) is highly prevalent in the elderly population. There are several types that call for specific treatment. The three most common options in managing UI are behavioral modification programs, pharmacologic agents, and surgical intervention. In general, a nonsurgical approach is the preferred first choice, unless it already has been tried and failed or if the patient clearly wishes a more definitive surgical therapy. PMID- 15341814 TI - Quality improvement and incontinence in long-term care. AB - The quality improvement framework provides a practical way for nursing homes to prevent and reduce urinary incontinence (UI)for residents. It provides a means for facilities to identify organizational factors that may be hindering appropriate continence care from being provided to residents. It enables nursing homes to integrate evidence-based UI practices and monitor the consistent application of those practices by staff. The regular collection and analysis of data related to UI is essential for identifying problems,seeking accurate solutions, providing staff with feedback regarding their performance, and evaluating the overall quality of continence care provided to residents. PMID- 15341815 TI - Psychosocial impact of urinary incontinence in the elderly population. AB - The impact of urinary incontinence extends well beyond the evident physical and physiologic sequelae. Incontinence may leave affected persons in social, emotional, and mental disarray. Measuring the psychosocial impact of a disease, however, is difficult,and there is no single best tool to achieve such an assessment. Several incontinence-specific tools have been devised to document the impact of this condition and are described briefly in this review. Outcome measures have, until recently, focused on objective data; however, from the patient's viewpoint, subjective psychosocial measures may be a better reflection of the success ofa treatment intervention. For a variety of reasons, the majority of affected persons do not seek help. If the full benefit of treatment options is to be realized, the health care provider actively should seek a history of incontinence in patients who may be ashamed or embarrassed. PMID- 15341816 TI - Urinary incontinence: a treatise on gender, sexuality, and culture. AB - Attitudes toward older adults with incontinence vary significantly between societies and cultures. Intangible consequences arising from such attitudes may exert enormous psychosocial strain on affected older adults, caregivers, family members, and social networks. Health providers should consider psychologic, gender based,and sociocultural differences in formulating efficient screening and therapeutic strategies for older adults who have UI. PMID- 15341817 TI - Geriatric fecal incontinence. AB - Fecal incontinence is a common problem in the elderly population,particularly in nursing homes, and is one of the common reasons for nursing home placement. In addition to the inconvenience of the incontinence for the patient and caregiver, it is associated with increased mortality. Identifiable physiologic changes in the anorectal region may contribute to the development of fecal incontinence.Fecal incontinence is a disorder of men and women, with an equal or greater prevalence in men in advancing years. All patients who have fecal incontinence warrant an initial medical evaluation,including the exclusion of fecal impaction. Cognitively impaired patients benefit most from habit training. Selected elderly patients who have fecal incontinence may benefit from biofeedback and surgical intervention. PMID- 15341819 TI - A question of balance. PMID- 15341820 TI - Palaeoecology, reference conditions and classification of ecological status: the EU Water Framework Directive in practice. AB - The European Union's Water Framework Directive (WFD) requires that all Member States within the European Union determine reference conditions for aquatic ecosystems to provide a baseline against which to measure the effects of past and present activities. Reference conditions are subsequently used to classify the ecological status of European waters. The decisions regarding environmental status will be important future elements in the management of European coastal waters. We have developed a number of classification scenarios for total nitrogen (TN) in the overlying waters of the southern part of Roskilde Fjord, Denmark, taking as our basis a palaeoecological reconstruction of fluctuations in TN between 1850 and 1995. We present a provisional classification scheme for the ecological status of Roskilde Fjord, sensu the WFD. Decision(s) regarding the deviation from reference conditions will give a wide range of apparent ecological status from good, through moderate and poor, to bad depending upon the definition of an acceptable deviation from reference conditions. The determination of an acceptable deviation will ultimately be a political decision, and will result in a wide range in the protection of coastal waters in Europe. There is still, however, an urgent need for a sound scientific documentation of the various scenarios for the implementation of the WFD. PMID- 15341821 TI - A comparison of neustonic plastic and zooplankton at different depths near the southern California shore. AB - Previous studies of neustonic debris have been limited to surface sampling. Here we conducted two trawl surveys, one before and one shortly after a rain event, in which debris and zooplankton density were measured at three depths in Santa Monica Bay, California. Surface samples were collected with a manta trawl, mid depth samples with a bongo net and bottom samples with an epibenthic sled, all having 333 micron nets. Density of debris was greatest near the bottom, least in midwater. Debris density increased after the storm, particularly at the sampling site closest to shore, reflecting inputs from land-based runoff and resuspended matter. The mass of plastic collected exceeded that of zooplankton, though when the comparison was limited to plastic debris similar to the size of most zooplankton, zooplankton mass was three times that of debris. PMID- 15341822 TI - Freezing of sediments inappropriate for pore water selenium analysis. AB - The common practice of freezing sediment cores for later chemical investigation was shown to be inappropriate for samples containing selenium. Pore waters from frozen estuarine sediment cores contained up to eighty times the selenium content of those extracted from chilled but unfrozen cores. Experiments suggested that the increase in selenium concentrations resulted from rupturing the cells of selenium-accumulating bacteria present in the samples. PMID- 15341823 TI - Distribution of DDT in sediments off the central California coast. AB - A collaborative sampling cruise off the central California coast was conducted to evaluate contaminant transport pathways along and across the shelf in the spring of 2002. The area has a complex current structure and net transport routes are not known for sure. Sediment characteristics, and organic and trace metal contaminants were analyzed in sediments taken from locations near shore, out to the heads of several canyons. Relative to the continental shelf and Pioneer Canyon stations, DDT was found at higher concentrations in Ascension and Monterey Canyons. Monterey Bay still receives DDT from terrestrial runoff and may be the source of DDT found in Ascension Canyon. DDT concentrations in Monterey Bay biota indicate bioaccumulation is occurring at depth due to continuing input from the shore. Effects on the deep ocean benthic community is unknown. PMID- 15341824 TI - Differential survival of coral transplants on various substrates under elevated water temperatures. AB - Closely related scleractinian coral species that exhibited similar survival patterns under relatively normal field conditions responded very differently to the occurrence of an environmental disturbance. The two species studied were Porites cylindrica and Porites rus which occur in the same reef zones in shallow reef flats. Transplants of both species were evenly distributed and attached to three different types of substrate: live coral colonies of P. cylindrica, dead coral colonies (also of P. cylindrica), and epoxy coated metal grids that were raised above the sandy substrate. With the onset of above-normal water temperatures due to the El Nino episode of 1998, P. cylindrica transplants immediately showed signs of bleaching stress and tissue necrosis, followed by algal overgrowth and mortality soon afterwards. In contrast, transplants of P. rus bleached more slowly and suffered less mortality, with a few actually showing signs of recovery at the end of the experimental period which covered a total of 14 weeks. These differences in responses could be attributed to properties of the symbiotic zooxanthellae, of the host coral tissue itself, or both. Over-all, survival was good on the metal grids (average of 35%), and on the live coral (average of 22%). It was poor on the dead coral (average of 6%). The metal grids as well as live coral tissue apparently provided a favorable substrate for the attached coral fragments, even for those of a different species. Under the conditions of this particular study, attachment of live coral fragments on already dead colonies for the purpose of increasing live coral cover on the reef did not yield favorable results. This is an area that requires further investigation. PMID- 15341825 TI - Ballast water sampling as a critical component of biological invasions risk management. AB - The human mediated transfer of harmful organisms via shipping, specifically via ballast water transport, leading to the loss of biodiversity, alteration of ecosystems, negative impacts on human health and in some regions economic loss, has raised considerable attention especially in the last decade. Ballast water sampling is very important for biological invasions risk management. The complexity of ballast water sampling is a result of both the variety of organism diversity and behaviour, as well as ship design including availability of ballast water sampling points. Furthermore, ballast water sampling methodology is influenced by the objectives of the sampling study. In the course of research conducted in Slovenia, new sampling equipment for ships' ballast water was developed and tested. In this paper new ballast water sampling methods and equipment together with practical shipboard testing results are presented. PMID- 15341826 TI - Spatial survey of trace metal contaminants in the sediments of the Elizabeth River, Virginia. AB - The Elizabeth River is a sub-estuary of the James River, the most southern tributary of the Chesapeake Bay. It is a highly industrialized area, and has been designated a "toxic hot spot" due to the heavy loads of contaminant metals and organic compounds in its bed sediments. Fifty surface sediment samples were taken along the channels and shoals of the Mainstem and the Southern Branch portions of the river. The samples were analyzed for trace metals (Cd, Cr, Cu, Pb and Zn), Fe and particle properties (% sand, % silt, % clay and specific surface area) to discern the spatial distribution of contaminant metals in this system. Enrichment factors were calculated to assess the overall level of contamination relative to other surrounding waterways as well as historic contamination levels within the river itself. The highest levels of metals were found to be in close proximity to industrial sources of contamination. The overall level of contamination was 3-10 times higher than baseline levels within the river, and 2-3 times higher than contaminant levels in other area waters. PMID- 15341827 TI - Sustainable impact of mussel farming in the Adriatic Sea (Mediterranean Sea): evidence from biochemical, microbial and meiofaunal indicators. AB - We have investigated the impact of a large mussel farm on the benthic environment using a battery of benthic indicators of environmental quality (including biochemical, microbial and meiofaunal parameters). These were analysed through a multi-control sampling strategy over one year. The differences across the seasons are typically higher than those between the impacted and the control stations. No effects are seen in terms of the sediment oxygen penetration and the downward fluxes (as the total mass, organic and phytopigment fluxes). The indicators based on the biochemical compositions of the sediment organic matter and the microbial parameters also show no evidence of the eutrophication process, except as a slight increase in the bacterial density in the sediments beneath the long-lines of the farm during the period of highest mussel stocks. Finally, no effects are observed in terms of the benthic faunal indicators, as the meiofaunal abundance, the community structure and the taxa richness are all indistinguishable between the farm sediments and the controls. These results show that mussel farming in the investigated system is eco-sustainable and does not significantly alter the coastal marine ecosystem, both in terms of the functioning and the trophic state. The battery of indicators selected in this study represents a useful tool for the monitoring of the potential ecological impact of mussel farms, towards guaranteeing the sustainable development of aquacultures in shallow coastal environments. PMID- 15341828 TI - Bacteriological examination of ballast water in Singapore Harbour by flow cytometry with FISH. AB - In this study the concentrations of total bacteria, enterobacteria, Vibrio spp., and E. coli have been compared for ballast water samples taken from ships in Singapore Harbour. The cell concentrations were enumerated using FISH and flow cytometry. The data were highly variable, reflecting the many influences upon ballast water as it is utilized in the shipping industry. The concentration of bacterial species was determined as a proportion of the total concentration of cells for the ballast water sampled. For the ballast water sampled these concentrations were 0.67-39.55% for eubacteria, 0-2.46% for enterobacteria, 0.18 35.82% for Vibrio spp., and 0-2.46% for E. coli. Using FISH and flow cytometry, an informative determination of the bacterial hazards of ship ballast water can be made. PMID- 15341829 TI - Nutrient enrichment caused by in situ fish farms at Eilat, Red Sea is detrimental to coral reproduction. AB - Recent studies report conflicting results concerning the effects of eutrophication on coral reproduction. The present study examines reproductive effort in the brooding coral Stylophora pistillata exposed to chronic eutrophication caused by in situ fish cages (FC) in the northern Gulf of Eilat (Aqaba). Histological studies of 20 S. pistillata colonies transplanted to each of two study sites, one close to the nutrient enriched FC site and the other at a reference site (IUI), 8 km southwest of the FC site, show that, overall, corals from the FC site have a significantly higher percentage of polyps containing oocytes and testes than corals from the IUI site. However, average oocyte size and the percentage of oocytes reaching the size at which fertilization occurs (i.e., >200 microm) were both significantly greater in colonies at the IUI site compared to the FC site. As the reproductive season progressed, colonies at the IUI site exhibited a decrease in the percentage of polyps containing oocytes, concomitant with an increase in the number of polyps containing planulae, indicating successful development of oocytes into planulae. In contrast, in colonies at the FC site oocyte numbers were greatest at the end of the reproductive season, and overall, numbers of planulae were significantly lower compared with the IUI colonies, suggesting relative failure of oocyte maturation, fertilization and ensuing larval development. The significantly higher lipid content found during the reproduction season in IUI colonies compared with FC colonies corroborates this assertion. This data strongly suggest that nutrients released from the fish farms have adverse effects on successful production of larvae of S. pistillata. In view of the recent severe deterioration of the coral reefs of Eilat and their present critical state of health, the only chance for their renewal is the use of immediate, prudent and rational protection measures against all man-made perturbations. PMID- 15341830 TI - Plastic debris ingestion by sea turtle in Paraiba, Brazil. AB - Coastal gill net entanglement and debris intake are important threats to the survival of sea turtles. Two sea turtles (Lepidochelys olivacea and Chelonia mydas) were found stranded along the coast of Paraiba. After necropsy, plastic debris were found in the stomach. The debris is described. This is the first record of this sort of problem for the Paraiba littoral. PMID- 15341831 TI - Levels and sources of polycyclic aromatic hydrocarbons in sediments from the Gulf of Suez. PMID- 15341832 TI - Identification and characterization of a new degradation product of Irgarol-1051 in mercuric chloride-catalyzed hydrolysis reaction and in coastal waters. PMID- 15341833 TI - Kinetic modeling of liquid-phase adsorption of phosphate on dolomite. AB - The adsorption of phosphate from aqueous solution on dolomite was investigated at 20 and 40 degrees C in terms of pseudo-second-order mechanism for chemical adsorption as well as an intraparticle diffusion mechanism process. Adsorption was changed with increased contact time, initial phosphate concentration, temperature, solution pH. A pseudo-second-order model and intraparticle diffusion model have been developed to predict the rate constants of adsorption and equilibrium capacities. The activation energy of adsorption can be evaluated using the pseudo-second-order rate constants. The adsorption of phosphate onto dolomite are an exothermically activated process. A relatively low activation energy and a model highly fitting to intraparticle diffusion suggest that the adsorption of phosphate by dolomite may involve not only physical but also chemisorption. This was likely due to its combined control of chemisorption and intraparticle diffusion. However, for phosphate/dolomite system chemical reaction is important and significant in the rate-controlling step, and for the adsorption of phosphate onto dolomite the pseudo-second-order chemical reaction kinetics provides the best correlation of the experimental data. PMID- 15341834 TI - Effects of pH and phosphate on the adsorptive fractionation of purified Aldrich humic acid on kaolinite and hematite. AB - The molecular weight (MW) fractionation of purified Aldrich humic acid (PAHA) resulting from adsorption on kaolinite and hematite was investigated for different solution pH and phosphate conditions. Adsorption was highly pH dependent, with higher uptake at lower pH values. For all pH conditions, the weight-average MW (MWw) of residual PAHA remaining in solution after adsorption deviated from the original MWw, indicating that preferential adsorption of certain MW components occurred. The extent of preferential adsorption depended on the percent carbon adsorption at a given pH condition. For similar percent carbon adsorption ranges, a greater extent of preferential adsorption of the higher MW PAHA components was observed with higher pH values as demonstrated by the lowest residual MWw value occurring at pH 9. Detailed analyses of selected residual PAHA samples clearly showed that adsorption selectivity for particular MW components was strongly influenced by solution pH. The extent of preferential adsorption of lower MW PAHA components decreased in the presence of a small amount of phosphate. This effect was more evident for hematite than kaolinite, and became greater with lower solution pH irrespective of the mineral type. The different fractionation patterns observed for PAHA were reasonably well explained by the physicochemical trends occurring in its MWw fractions and the underlying sorption processes. PMID- 15341835 TI - Impact of humate complexation on the adsorption of REE onto Fe oxyhydroxide. AB - Adsorption experiments of rare-earth elements (REE) onto hydrous ferric oxide (HFO) were performed to evaluate the impact of organic complexation on both REE(III) adsorption and the Ce(III) oxidation rate. Scavenging experiments were performed at pH 5.2 with NaCl and NaNO3 solutions containing either free REE (III) or REE(III)-humate complexes. The log K(d)(REE) patterns obtained from HFO suspensions exhibit a slight positive Ce anomaly and an M-type lanthanide tetrad effect, in contrast with the partitioning between REE(III)-humate complexes and HFO, which yields completely flat distribution patterns. The "organic" partitioning runs yield log K(d)(REEorganic)/log K(d)(DOC) ratios (DOC = dissolved organic carbon) close to 1.0, implying that the REE(III) and humate remain bound to each other during the adsorption experiment. The lack of any positive Ce anomaly or M-type lanthanide tetrad effect in the organic experiments seems to reflect an anionic adsorption of the REE-humate complex. Adsorption onto HFO takes place via the humate side of the REE(III)-humate complexes. The oxidation of Ce(III) by Fe(III) and the proportion of surface hydroxyl groups coordinated to REE(III) at the HFO surface are the two most commonly invoked processes for explaining the development of positive Ce anomalies and the M-type tetrad lanthanide effect. However, such processes cannot proceed since the REE are not in direct contact with the HFO suspensions, the latter being shielded by PHA. The present results further complicate the use of Ce anomalies as reliable paleoredox proxies in natural precipitates. They are also further demonstration that organic matter may inhibit the lanthanide tetrad effect in geological samples. PMID- 15341836 TI - Adsorption properties of gases on mesoporous chromium silicate. AB - The properties of nitrogen, carbon dioxide, and nitrogen dioxide adsorption onto mesoporous chromium silicates were studied by measurements of both the adsorption isotherms and the IR spectra. The pore sizes of two types of chromium silicates, Cr-FSM-16 (Si/Cr=170 (Cr-FSM-16 [170]) and 390 (Cr-FSM-16 [390])), which contain different amount of Cr, were 2.75 nm. BET surface areas of Cr-FSM-16 were 590 m2/g and they were smaller than that onto FSM-16. The initial heat of adsorption of nitrogen onto Cr-FSM-16 was higher than that onto FSM-16. But the initial heat of adsorption of carbon dioxide onto Cr-FSM-16 was smaller than that onto FSM-16. These results indicated that Cr in Cr-FSM-16 decreased adsorption interaction with carbon dioxide. When nitrogen dioxide was adsorbed on FSM-16 and Cr-FSM-16 at 303 K under no light, an absorption band of nitrogen monoxide adsorbed was measured by IR spectroscopy. This decomposition of nitrogen dioxide by FSM-16 and Cr-FSM-16 was caused by SiOH and Cr, respectively. PMID- 15341837 TI - Bovine serum albumin (BSA) plays a role in the size of SDS micelle-like aggregates at the saturation binding: the ionic strength effect. AB - The influence of ionic strength on the complexes formed by natural bovine serum albumin (BSA), pH 5.4 (near the isoelectric point), and sodium dodecyl sulfate (SDS) in aqueous buffered (sodium acetate) solution was investigated by using surface tension, fluorescence and small angle X-ray scattering (SAXS) techniques. Ionic strength was varied by changing sodium acetate buffer concentration from 0.020 to 0.5 M. Surface tension revealed that SDS:BSA saturation binding occurs at psp = 42 +/- 2 mM, independent of the solution ionic strength. Further, SAXS curves are consistent with the necklace and bead model, where micelle-like aggregates are randomly distributed along the partial unfolded protein. Micelle like aggregates grow from small spheres at 10 mM SDS to small ellipsoids (upsilon = 1.3 , ratio between the largest and the shortest axes) near psp, in good agreement with micellar aggregation numbers obtained by fluorescence, independent of salt concentration. Protein-bound micelles stop growing above psp and further SDS addition induces free-micelle formation. PMID- 15341838 TI - Adsorption kinetics of zinc in multicomponent ionic systems. AB - Using commercial activated carbon as an adsorbent, the kinetics of adsorption of zinc from multicomponent ionic systems having cadmium and mercury has been studied and reported. The variables investigated have been the chemical nature, ionic strength, and pH of the adsorptive (Zn2+) solution. The adsorption of Zn2+ is speeded up by the presence of Cd2+ and Hg2+ ions provided that the concentration of these two ions is high as compared to the concentration of Zn2+. When the ionic strength of the solution in relative terms is high (i.e., > 3 x 10(-4) M), however, the adsorption of Zn2+ decelerates. Also, the adsorption process is greatly accelerated at pH 12, whereas at pH 2 it does not occur at all. PMID- 15341839 TI - Sodium chloride-induced self-assembly of microfibers from nanofiber components. AB - Self-assembly of a nucleotide bolaamphiphile, terminated with two nucleotide moieties including thymine, deoxyribose, and phosphodiester at both ends of the C20 oligomethylene chain, in sodium chloride aqueous solutions has been studied by SEM, LSM, FE-SEM, micro-FTIR, and zeta-potential measurement. The self assembly behavior of the nucleotide bolaamphiphile was strongly dependent on the concentration of sodium chloride added. The nucleotide bolaamphiphile was found to hierarchically self-assemble to form micrometer-sized fibers (microfibers) consisting of bundles of entangled nanometer-sized fibers (nanofibers) under certain conditions of sodium chloride concentration (approximately 44 mM). The zeta-potential measurement suggested that the surface-charge tuning of the nanofibers induce the hierarchical self-assembly of the microfibers. PMID- 15341840 TI - Synthesis and surface properties of aqueous dispersions of poly(ester-imide) prepared from anhydride terminated polyester prepolymer and diisocyanate. AB - Aqueous dispersions of poly(ester-imide)s [P(E-I)s] have been prepared by dispersing the P(E-I)s in water without any external solubilizing agents. P(E-I)s were prepared from anhydride-terminated polyester prepolymer and diisocyanate. The -COOH groups in the polymer were then neutralized using triethylamine and the P(E-I)s were subsequently dispersed in water. The influence of the degree of ionization of polymers on the particle size and viscosity of the dispersion has been studied. The dispersions were crosslinked using polyaziridine. The crosslinked dispersion cast films were characterized for dynamic mechanical properties. As the ionic content increased the particle size decreased and the viscosity increased. When the amount of crosslinker added was varied, for a fixed percentage of ionization, the glass transition temperature Tg shifted to higher values. Critical surface tension (CST) measurements indicated reorganization of hydrophobic groups on the surface after crosslinking. PMID- 15341841 TI - Synthesis and dispersion of Ni(OH)2 platelet-like nanoparticles in water. AB - Synthesis of nanometric platelet-like Ni(OH)2 particles is described. The role of several experimental parameters on the particle size is investigated. A colloidal dispersion of particles is produced by adsorbing ionizable organic ligands (trisodium citrate) on the particle surface. The stability of this colloidal dispersion and the particle charge density are determined for different citrate ions concentrations. PMID- 15341842 TI - Investigations on the semi-batch precipitation of silica. AB - The semi-batch precipitation process of silica from sodium silicate and sulphuric acid has been structured into a primary particle production stage, an aggregation process until the gel point is reached and finally a gel fragmentation and compaction process. The progress of aggregation was analyzed by photon correlation spectroscopy. The reaction limited aggregation kernel was found to be proportional to B'(t)(q(p - q))A/2, where A assumes the value of one. The quantity B'(t) varies between 3.4 x 10(-20) and 2.6 x 10(-20) 1/min and is only a weak function of time. The stability ratio W for the aggregation process decreases with increasing aggregate size. During the period after the gel point the gel fragmentates. The gel clusters have an initial fractal dimension of approximately 1.8 which rapidly increases to 2.4 and stays roughly constant until the end of the process. The specific surface area decreases from 480 to 300 m2/g. The primary particles have a uniform size of 22.7 nm. PMID- 15341843 TI - Reverse Monte Carlo modeling of the structure of colloidal aggregates. AB - In this work we present results for the structure of aerogels coming from the diffusion-limited cluster aggregation simulation method. Pair distribution functions and structure factors, resulting from simulation, were considered as experimental input for reverse Monte Carlo modeling. The modeling yielded structural models with pair distribution functions and structure factors nearly identical to the results of the simulations. Particle configurations from both the simulations and reverse Monte Carlo modeling have been analyzed in terms of the distribution of the number of neighbors. It is suggested that the reverse Monte Carlo method, when applied to the structure factor, may be a suitable technique for the interpretation of experimental scattering data on colloidal aerogels. PMID- 15341844 TI - Temperature dependence of particle-particle adherence forces in ice and clathrate hydrates. AB - Particle-particle pulloff adherence forces were measured as a function of temperature in the ice/n-decane/ice and tetrahydrofuran (THF) hydrate/n decane/THF hydrate systems using a newly developed micromechanical testing technique. Experiments using approximately 200 microm radius particles were performed at atmospheric pressure over the temperature range 263-275 K. The ice and hydrate particles displayed very similar behavior. While the measured adherence forces had significant variation, the shapes of the cumulative force distribution curves were similar among the different sets of experiments. The measured adherence forces distributions shifted to lower force values as the temperature was decreased from the solid melting temperature. The observed forces and trends were explained by the capillary cohesion of rough surfaces, with the capillary bridging liquid being stabilized below its freezing point by the negative curvature of the bridging liquid/n-decane interface. PMID- 15341845 TI - Surface modified ormosil nanoparticles. AB - Organically modified silanes (ORMOSIL) such as vinyl triethoxysilane readily aggregate in the aqueous cores of reverse micelles where the triethoxysilane moieties are hydrolyzed to form a hydrated silica network and the vinyl groups protruded out from the surface of the nanoparticles toward the hydrophobic side of the micellar interface. These particles are spherical and the size distribution of the particles is relatively narrow, with an average diameter of 87 nm. Surface vinyl silica nanoparticles so formed have been oxidized to surface carboxylic silica nanoparticles, followed by chemical conjugation with polyethyleneglycol amine (PEG amine) through the ethyl-3-(3-dimethylaminopropyl) (EDCI) carbodiimide reaction. The characteristic surface groups have been identified by Fourier transform infrared spectroscopy, while the size and the morphology of the particles have been studied by dynamic light scattering and transmission electron microscopy. It has been found that about 80-85% of the carboxylic groups are PEGylated during the EDCI reaction. PMID- 15341846 TI - Electroacoustic theory for concentrated colloids with overlapped DLs at arbitrary kappa alpha. I. Application to nanocolloids and nonaqueous colloids. AB - Existing theories of electroacoustic phenomena in concentrated colloids neglect the possibility of double layer overlap and are valid mostly for the "thin double layer," when the double layer thickness is much less than the particle size. In this paper we present a new electroacoustic theory which removes this restriction. This would make this new theory applicable to characterizing a variety of aqueous nanocolloids and of nonaqueous dispersions. There are two versions of the theory leading to the analytical solutions. The first version corresponds to strongly overlapped diffuse layers (so-called quasi-homogeneous model). It yields a simple analytical formula for colloid vibration current (CVI), which is valid for arbitrary ultrasound frequency, but for restricted kappa alpha range. This version of the theory, as well the Smoluchowski theory for microelectrophoresis, is independent of particle shape and polydispersity. This makes it very attractive for practical use, with the hope that it might be as useful as classical Smoluchowski theory. In order to determine the kappa alpha range of the quasi-homogeneous model validity we develop the second version that limits ultrasound frequency, but applies no restriction on kappa alpha. The ultrasound frequency should substantially exceed the Maxwell-Wagner relaxation frequency. This limitation makes active conductivity related current negligible compared to the passive dielectric displacement current. It is possible to derive an expression for CVI in the concentrated dispersion as formulae inhering definite integrals with integrands depending on equilibrium potential distribution. This second version allowed us to estimate the ranges of the applicability of the first, quasi-homogeneous version. It turns out that the quasi-homogeneous model works for kappa alpha values up to almost 1. For instance, at volume fraction 30%, the highest kappa alpha limit of the quasi homogeneous model is 0.65. Therefore, this version of the electroacoustic theory is valid for almost all nonaqueous dispersions and a wide variety of nanocolloids, especially with sizes under 100 nm. PMID- 15341847 TI - The effect of annealing temperature on latex film dissolution. AB - The steady-state fluorescence technique (SSF) was used to study the dissolution of polystyrene (PS) latex films in toluene. The main objective of this study was to explore the effects of annealing temperature on the dissolution behavior of the PS films. It has been shown that dissolution coefficients of PS films are highly affected by the annealing temperatures. It was found that, while PS films dissolved as entangled clusters when annealed at low temperatures, they dissolved as individual chains at high annealing temperatures. The results showed that when the annealing temperature increases, the dissolution takes place in the disentanglement regime. The decrease in dissolution coefficients, Dd, can be explained as a shift of dissolution from the entangled clusters regime to the disentanglement regime. The measured Dd values were found to be in between 4.2 and 1.0 x 10(-10) cm2 s(-1) for an annealing temperature range of 110-220 degrees C. PMID- 15341848 TI - Uranyl sorption by smectites: spectroscopic assessment of thermodynamic modeling. AB - Batch sorption experiments and thermodynamic modeling of the interaction of UO2(2+) and its hydrolysis products with two smectitic clay minerals, the reference material SWy-1 [McKinley et al., Clays Clay Miner. 43 (1995) 586] and the soil isolate LK-1 [Turner et al., Geochim. Cosmochim. Acta 30 (1996) 3399], have established a conceptual framework for uranyl/smectite surface complexation based on general reactions between aqueous uranyl species and the reactive sites on the mineral surfaces. In this report, we have formulated and spectroscopically tested a set of hypotheses based on this conceptual framework using samples prepared under similar or identical conditions to evaluate the agreement between surface complexation/speciation as enumerated by spectroscopic characterization and that elaborated by the surface complexation model. Both steady-state and time resolved optical emission spectral data are presented for uranyl on both smectite minerals as well as on the analogue phases SiO2 and Al(OH)3 spanning the pH range from approximately 4 to 8 and the background electrolyte concentrations from approximately 0.001 to 0.1 M. The spectral data enable the explicit identification of an outer-sphere exchange-site population of the hydrated cation [UO2(OH2)5(2+) ] in SWy-1. Spectral data also clearly establish the existence of inner-sphere surface complexes on the analogue phases and on the amphoteric clay crystallite edge sites [aluminol (>Al-OH) and silanol (>Si-OH)]. Based on the spectral characteristics of these uranyl edge-site populations, it is possible to readily infer for the SiO2, Al(OH)3, and SWy-1 samples the evolution in surface speciation with increasing pH to more hydrolyzed uranyl-surface complexes consistent with the conceptual model. The spectral domain characteristics of the edge-site populations on LK-1 with increasing pH suggest that there is no change in the hydrolysis of the uranyl-surface species. However, emission lifetime data are interpreted as indicating a shift in the surface speciation of the same uranyl-surface species from aluminol sites to silanol sites with pH increase. This observation is also consistent with the conceptual framework of the model. Data are also reported for Eu3+/smectite samples to provide additional insight into the exchange site populations. The emission spectra for Eu3+ in the basal plane exchange sites differs significantly between SWy-1 and LK-1 samples reflecting a difference in the basal plane spacing between these two minerals, but the emission lifetime data suggest that the Eu3+ cation remains fully hydrated in both systems. The overall general description of surface speciation of uranyl on these mineral phases as enumerated by spectroscopy is in good accord with that derived from the conceptual thermodynamic model, lending added confidence to our understanding and descriptions of surface complexation behavior in this complex geochemical system. PMID- 15341849 TI - Calorimetry by immersion into liquid nitrogen and liquid argon: a better way to determine the internal surface area of micropores. AB - The aim of this work is to assess the internal surface area of a set of samples (either carbons or oxides, either porous or nonporous, either microporous or mesoporous) by microcalorimetry via immersion into liquid nitrogen or argon. We have made use of an isothermal, heat-flux microcalorimeter, initially designed and built in our laboratory for the sake of gas adsorption experiments at 77 or 87 K. It seems that immersion calorimetry into liquid nitrogen and argon makes it possible to go one step further in the determination of the internal surface area of micropores. PMID- 15341850 TI - Advective flow in a sludge floc. AB - The interior of sludge floc is highly heterogeneous, while the large pores in the floc control the advective flow. This work for the first time numerically details fluid flow and mass transfer processes in pores of activated sludge floc. The dimensionless permeabilities and mass dispersion coefficients were contoured against pore size ratio and the floc Reynolds number. With a pore size less than 20% of the floc size, the commonly adopted homogeneous model overestimates the floc permeability, and pore velocity is less than 2% of the bulk velocity. This is particularly true for flocs with low porosity. Although the convective flux is low, the dispersive mass transfer rate can be much higher than the diffusional rate, attributable to the strong Taylor dispersion effect. The three-dimensional pore structures in waste activated-sludge floc were identified using confocal laser scanning microscope (CLSM) images. Large pores were used to numerically estimate the permeability and dispersion coefficient for these pores. The permeability and the dispersion coefficient of the tortuous pores can be one order of magnitude lower than those for the equivalent straight pores. Besides the dispersion effect, the pore tortuosity appeared as the most important geometrical factor retarding the advective flow in the sludge pores. In addition, the small side pores connected to the large pore had only a mild effect on the flow process, and can be neglected in analysis. PMID- 15341851 TI - The pH stimulated reversible loading and release of a cationic dye in a layer-by layer assembled DNA/PAH film. AB - Through the layer-by-layer (LbL) deposition method, DNA was assembled into an ultrathin film with a cationic poly(allylamine hydrochloride) (PAH). The loading and release of a typical cationic dye, 5, 10, 15, 20-tetrakis(4-N methylpyridyl)porphine-tetra-(p-toluenesulfonate) (TMPyP), in the DNA/PAH films were investigated. It has been found that the LbL-assembled DNA/PAH film was very stable in both acidic and alkaline solutions. Stimulated by the pH change of the dye solution, the dye can be easily loaded into or released from the DNA/PAH film. In an alkaline solution, the dye could be rapidly loaded into the DNA/PAH film at room temperature, while in an acidic solution, the dye could be rapidly released. The mechanism of such pH-stimulated loading and release in the DNA/PAH film was discussed. It was further observed that the loading and release of the dye in the DNA/PAH film was reversible upon pH change and the process could be repeated many times. PMID- 15341852 TI - Equilibrium partitioning of Ficoll in composite hydrogels. AB - Equilibrium partition coefficients (phi, the concentration in the gel divided by that in free solution) of fluorescein-labeled Ficolls in pure agarose and agarose dextran composite gels were measured as a function of gel composition and Ficoll size. The four narrow fractions of Ficoll, a spherical polysaccharide, had Stokes Einstein radii ranging from 2.7 to 5.9 nm. Gels with agarose volume fractions of 0.040 and 0.080 were studied, with dextran volume fractions (calculated as if the chain were a long fiber) up to 0.011. As expected, phi generally decreased as the Ficoll size increased (for a given gel composition) or as the amount of dextran incorporated into the gel increased (for a given agarose concentration and Ficoll size). The decrease in phi that accompanied dextran addition was predicted well by an excluded volume theory in which agarose and dextran were both treated as rigid, straight, randomly positioned and oriented fibers. Modeling dextran as a spherical coil within a fibrous agarose gel produced much less accurate predictions. The diffusional permeabilities of these gels were assessed by combining the current partitioning data with relative diffusivities (Kd, the diffusivity in the gel divided by that in free solution) reported previously. The values of phi Kd for a synthetic gel with 8.0% agarose and 1.1% dextran (by volume) were found to be very similar to those for the glomerular basement membrane, a physiologically important material which also has a total solids content of approximately 10%. PMID- 15341853 TI - Fractal analysis of hydroxyapatite from nitrogen isotherms. AB - Samples of calcium hydroxyapatite, CaHap, were prepared via a wet method and subjected to thermal treatment in air in the temperature range 100-900 degrees C. Nitrogen adsorption-desorption isotherms were obtained on different samples, and their data points were used to analyze the fractal properties of the obtained solids. Both FHH and Neimark's equations were used for such purpose, and the agreement or disagreements between obtained results on using both equations are discussed. Considering the appropriate values of the obtained surface fractal dimensions D, it was concluded that an appreciable defractalization of the prepared hydroxyapatite is only noted upon calcinations at 900 degrees C. Below this temperature the loss in surface area and pore volume result from simultaneous bulk and surface mass transport, which conserve the initial surface roughness and average pore radius. Calcination at 900 degrees C caused sintering to proceed via bulk mass transport, with a consequent pore widening and a decrease in surface roughness. PMID- 15341854 TI - Intraparticle phosphorus diffusion in a drinking water treatment residual at room temperature. AB - Phosphorus (P) has been recognized as one of the major limiting nutrients that are responsible for eutrophication of surface waters, worldwide. Efforts have been concentrated on reducing P loads reaching water bodies, via surface runoff and/or leaching through a soil profile. Use of drinking water treatment residuals (WTRs) is an emerging cost-effective practice to reduce soluble P in poorly P sorbing soils or systems high in P. Literature suggests that WTRs have huge P sorption capacities. We hypothesized that P sorption would be limited by diffusional constraints imposed by the WTR particles. Selected chemical and physical (specific surface area, particle size distribution) characteristics of an iron-based WTR were measured. Sorption P isotherms at room temperature were constructed, and sorption kinetics were monitored. An intraparticle diffusion model was utilized to fit the kinetic data. Results showed that the WTR dramatically reduced soluble P, showing nonequilibrium characteristics, even after 80 d of reaction. Specific surface area (SSA) measured with CO2 gas was significantly greater than the traditional BET-N2 value (28 versus 3.5 m2 g(-1)), suggesting that a large amount of internal surfaces might be present in the WTR. The intraparticle P diffusion model was modified to include the wide particle size distribution of the WTR. The intraparticle diffusion model fitted the data well (r2 = 0.83). We calculated a maximum apparent P diffusion coefficient value of 4 x 10(-15) cm2 s(-1), which agrees with published values for intraparticle diffusion in microporous sorbents. This work may be useful for predicting long term sorption characteristics of WTRs, since WTRs have been suggested as potential long-term immobilizers of sorbed P in P-sensitive ecosystems. PMID- 15341855 TI - Dynamics of water spreading on a glass surface. AB - The impact of a water droplet on a glass surface is studied experimentally using a high-speed video camera which can catch up to 60,000 images per second with an exposure time of 10 micros. A wide range of impact velocities are studied by varying the fall height, showing different spreading regimes. Particular attention is given to the dynamics of the contact angle and its relation to the maximum expanding radius and capillary number. A linear relation between the contact line velocity and the impact velocity is found experimentally. Using acoustic analysis, an evaluation of the pressure at the contact line is given. We also confront predicted and experimental jetting times. This work shows that descriptions of drop impact based purely on conservation of energy are inadequate to describe the dynamics of the event. The different shapes taken by the drops between the initial impact and the maximum radius determine the final outcome. PMID- 15341856 TI - Determination of the critical surface tension of wetting of minerals treated with surfactants by shear flocculation approach. AB - This paper contributes the shear flocculation method as a new approach to determine the critical surface tension of wetting of minerals treated with surfactants. This newly developed approach is based on the decrease of the shear flocculation of the mineral suspension, with decreasing of the surface tension of the liquids used. The solution surface tension value at which shear flocculation does not occur can be defined as the critical surface tension of wetting (gamma c) of the mineral. By using the shear flocculation method, the critical surface tensions of wetting (gamma c) for calcite and barite minerals, treated with surfactants, were obtained as 30.9 and 35.0 mN/m, respectively. These values are in good agreement with data reported previously on the same minerals obtained by the contact angle measurement and flotation methods. The chemical agents used for the treatment of calcite and barite particles were sodium oleate and sodium dodecyl sulfate, respectively. PMID- 15341857 TI - Dynamics of nonionic surfactant-rich phase separation and recovery of dyes. AB - Cloud point separation of selected dyes was studied. The use of dyes made possible observation of the dynamics of surfactant-rich phase separation by color video. The pictures were interpreted by means of ImageC software and degrees of whiteness were calculated. It was found that separation was slow and equilibrium was not achieved even after a period of more than 10 h. The separated surfactant rich phase had a heterogeneous structure. The globules of the surfactant-rich phase were also observed in the micellar aqueous phase. The surfactant concentration could be as high as one or two orders of magnitude above the expected cmc values and was not decreased to critical micelle concentration by centrifugation. The presence of sodium chloride was important and improved separation. Separation of dyes was in the range 73-98% and depended upon the surfactant, the temperature, the electrolyte content, and the dye. In each system considered, appropriate conditions had to be selected to obtain high recovery of the dye. The presence of the electrolyte was the most important parameter and it improved the separation of dyes. However, addition of the electrolyte could also account for precipitation of the dyes, as observed for the systems containing Direct Pink. The best recoveries were observed for Direct Yellow and oxyethylated nonylphenol (98% at 55 degrees C in the presence of NaCl). Centrifugation gave recoveries similar to those for prolonged heating but it shortened the time of phase separation. PMID- 15341858 TI - Synthesis, characterization, and SANS studies of novel alkanediyl-alpha,omega bis(hydroxyethylmethylhexadecylammonium bromide) cationic gemini surfactants. AB - A series of novel gemini cationic surfactants alkanediyl-alpha,omega-bis (hydroxyethylmethylhexadecylammonium bromide) with polymethylene spacer chain length of 4, 6, 8, and 10 carbon atoms was synthesized and characterized. Critical micellar concentrations of the gemini surfactants in aqueous solutions as determined by the surface tension and conductance measurements were observed to be in the range 1.39-3.63 microM. The critical micellar concentration was observed to increase initially with spacer length up to 6 methylene groups and to decrease thereafter with the increase in spacer length. The micellar microstructure in aqueous solutions examined through small angle neutron scattering (SANS) revealed that the extent of aggregation growth and variation in shapes of micelles strongly depend on head group polarity, spacer chain length, and temperature. The propensity to micellar growth with spacer chain length 4 was found to be much higher than with the longer spacer lengths. The fractional charge on the micelle increases with increased spacer chain length and temperature. PMID- 15341859 TI - Influence of dynamic interfacial tension on droplet formation during membrane emulsification. AB - Membrane emulsification is a promising and relatively new technique for producing emulsions. The purpose of this study was to better understand the influence of interfacial tension on droplet formation during membrane emulsification. Droplet formation experiments were carried out with a microengineered membrane; the droplet diameter and droplet formation time were studied as a function of the surfactant concentration in the continuous phase. These experiments confirm that the interfacial tension influences the process of droplet formation; higher surfactant concentrations lead to smaller droplets and shorter droplet formation times (until 10 ms). From drop volume tensiometer experiments we can predict the interfacial tension during droplet formation. However, the strong influence of the rate of flow of the to-be-dispersed phase on the droplet size cannot be explained by the predicted values. This large influence of the oil rate of flow is clarified by the hypothesis that snap-off is rather slow in the studied regime of very fast droplet formation. PMID- 15341860 TI - Characterization and demulsification of poly(ethylene oxide)-block-poly(propylene oxide)-block-poly(ethylene oxide) copolymers. AB - Four poly(ethylene oxide)-block-poly(propylene oxide)-block-poly(ethylene oxide) copolymers with different molecular weights and PPO/PEO composition ratios were synthesized. The characterization of the PEO-PPO-PEO triblock copolymers was studied by surface tension measurement, UV-vis spectra, and surface pressure method. These results clearly showed that the CMC of PEO-PPO-PEO was not a certain value but a concentration range, in contrast to classical surfactant, and two breaks around CMC were reflected in both surface tension isotherm curves and UV-vis absorption spectra. The range of CMC became wider with increasing PPO/PEO composition ratio. Surface pressure Pi-A curves revealed that the amphiphilic triblock copolymer PEO-PPO-PEO molecule was flexible at the air/water interface. We found that the minimum area per molecule at the air/water interface increased with the proportion of PEO chains. The copolymers with the same mass fractions of PEO had similar slopes in the isotherm of the Pi-A curve. From the demulsification experiments a conclusion had been drawn that the dehydration speed increased with decreased content of PEO, but the final dehydration rate of four demulsifiers was approximate. We determined that the coalescence of water drops resulted in the breaking of crude oil emulsions from the micrograph. PMID- 15341861 TI - Small-angle X-ray scattering and electron paramagnetic resonance study of the interaction of bovine serum albumin with ionic surfactants. AB - Small-angle X-ray scattering (SAXS) and electron paramagnetic resonance (EPR) techniques have been used to monitor the interaction of bovine serum albumin (BSA) with ionic surfactants such as anionic sodium dodecyl sulfate (SDS), zwitterionic N-hexadecyl-N,N-dimethyl-3-ammonium-1-propane sulfonate (HPS), and cationic cethyltrimethylammonium chloride (CTAC) at pH 7.0. The SAXS results have shown that in the presence of 5 mM SDS and HPS the radius of gyration (Rg) almost does not change as compared to the BSA free-surfactant solution; its value is ca. 30 Angstroms. In the presence of 5 mM CTAC the SAXS data indicate the presence of a particle with a Rg of at least 63 Angstroms, suggesting that in this case, a kind of protein aggregation takes place. In the presence of SDS and HPS surfactants at concentrations above 10 mM, a characteristic broad peak in the region of 0.12-0.18 Angstroms(-1) indicates the presence of micelle-like aggregates in solution. The SAXS curves are consistent with the "pearl necklace" model, where micelle-like aggregates are randomly distributed around the polypeptide chain. EPR results using 5-DSA and 16-DSA spin labels show that in the presence of BSA the EPR spectra are composed of two label populations, one contacting the protein and a second one due to label localization in the micelles. Evidence is also obtained for a competition of the surfactants with the spin labels for the high-affinity binding sites of the stearic acid spin labels as monitored by changes in the fractions of the two label populations as the surfactant concentration is increased. The effect of SDS seems to be stronger in the sense that increased SDS concentration leads to a complete transfer of spin labels from close protein contact sites to micelles, while for HPS, a significant immobilization of probe apparently remains even at higher surfactant concentrations. These two techniques are quite useful since SAXS monitors the overall properties of the scattering particle, while EPR gives information on the dynamics inside this particle and associated with label localization and motion. PMID- 15341862 TI - Molecular dynamics simulation studies of the transport and adsorption of a charged macromolecule onto a charged adsorbent solid surface immersed in an electrolytic solution. AB - Molecular dynamics simulations were performed in order to study the transport and adsorption of a charged macromolecule (desmopressin) onto a charged solid surface in an electrolytic solution. The strong Coulombic interaction from the charged solid surface represents the major force for accelerating, orienting, entrapping in the electrical double layer, and adsorbing the macromolecule onto the charged solid surface. The macromolecule is flattened as it approaches the charged surface, giving rise to a stronger surface exclusion effect that shields surface sites. When adsorbed, the macromolecule is restrained by a surface interaction more than one hundred times stronger than the thermal energy, of which 99.8% results from the strong dominant Coulombic interaction, and trapped by a hydration layer adjacent to the surface. This leads to zero lateral displacement of the adsorbed macromolecule and indicates that surface diffusion is a physically implausible mechanism in similar systems. Explicit solvent is required for realistic representation of the macromolecular structure and the surface interaction energy. The adsorbed macromolecule also decreased the electrostatic potential gradient perpendicular to the charged solid surface and introduced additional electrostatic potential gradients laterally. The results obtained from the molecular dynamics simulations confirm the importance of electrophoretic migration and support the physical mechanisms used in a macroscopic continuum model that predicts an overshoot in the concentration of a charged macromolecule in the adsorbed phase under certain conditions of pH and ionic strength. PMID- 15341863 TI - Modification of indium-tin oxide (ITO) glass with aziridine provides a surface of high amine density. AB - The surface of indium-tin oxide (ITO) substrates was successfully modified with aziridine. Modification of the surface was achieved through facile ring-opening, and hyperbranching polymerization of the ring-strained heterocycle initiated from the reactive group on the surface. Amine density of the aziridine-modified ITO measured with UV-vis spectrophotometry is 10 amines/nm2. Cyclic voltammetric analysis showed that the aziridine-modified electrode was less active for Ru(NH3)6(3+) in comparison with the pristine electrode, while no difference was observed for Fe(CN)6(4-). Electrochemical impedance spectroscopic experiments unveiled that the modified electrode was more efficient for electron transfer to the latter species than to the former. PMID- 15341864 TI - New understanding of ischemic mitral regurgitation: the marionette and its masters. PMID- 15341865 TI - Left ventricular Tei index in athletes. PMID- 15341867 TI - Report on the first written exam held as part of the European Association of Echocardiography accreditation process in adult transthoracic echocardiography. PMID- 15341868 TI - Echocardiographic classification of chronic ischemic mitral regurgitation caused by restricted motion according to tethering pattern. AB - Although the mechanism of ischemic mitral regurgitation (MR) is understood, the echocardiographic picture of ischemic MR is not homogeneous. Ninety-two consecutive patients with chronic ischemic MR due to restricted motion were divided into two groups according to tethering pattern: the asymmetric group with predominant posterior tethering of both leaflets (54 patients) and the symmetric one with predominant apical tethering of both leaflets (38 patients). The mitral deformation indexes, LV global (volume, function and sphericity) and local (papillary muscle displacements and regional wall motion score index) remodeling were evaluated. All indexes of global LV remodeling were significantly higher in the symmetric than asymmetric group (all p < 0.0001), such as the posterior and lateral displacement of the anterior papillary muscle (both p < 0.04), the papillary muscle separation and the anterior papillary muscle wall motion index (both p < 0.0001). The origin as well as the direction of the jet was central in all patients of the symmetric group. In the asymmetric one the origin was central in 78% of the cases and arising from the medial commissure in 22% whereas the jet direction was posterior and central in 83% and 17% of patients, respectively. Therefore, it is possible to distinguish at least two subgroups of patients with ischemic MR due to restricted motion on the basis of tethering pattern, different degree of local and global LV remodeling and characteristics of the regurgitant jet. PMID- 15341869 TI - Factor analysis of the left ventricle by echocardiography (FALVE): a new tool for detecting regional wall motion abnormalities. AB - BACKGROUND: Factor analysis of the left ventricle in echography was developed to study the regional wall motion. Two factors and associated factor images were estimated using specific constraints: one "constant" factor and another "contraction-relaxation" factor. The constant factor was encoded in green, the positive component of the contraction in red and the negative in blue. METHODS: The evaluation was carried out on 12 patients with LBBB or pacemaker (group A), and on 26 others (group B). The segments were graded separately on the cine-loops by three experienced echocardiographers. Similarly, the three-color combination of the factor images was read at the endocardial border and each segment was scored. RESULTS: An absolute concordance was obtained for 64.8% of the segments and a relative concordance (within one grade) for 97.2%. They were 71% and 99.6% in group B. Most of the discordant cases were explained by the global motion during the cardiac cycle. The standard deviation of the difference between the mean wall motion scores was 0.38 for all the patients; it was reduced to 0.30 in group B. CONCLUSION: Factor analysis is a promising tool to study the regional wall motion. It might become useful for assessing segmental wall motion in 2D and 3D echo. PMID- 15341870 TI - Echo-guided percutaneous septal ablation for symptomatic hypertrophic obstructive cardiomyopathy: 7 years of experience. AB - AIMS: To analyze the impact of intra-procedural echocardiographic imaging on the interventional strategy in ethanol-induced septal ablation (PTSMA) for symptomatic hypertrophic obstructive cardiomyopathy (HOCM), based on a single center experience of 7 years. METHODS AND RESULTS: PTSMA was intended for refractory symptoms in 337 patients (pts.) with HOCM (mean age: 54 +/- 15 years), with 312 procedures completed by injection of 2.8 +/- 1.2 ml ethanol. In 25 pts. (8%) the intervention was aborted without ethanol injection, mostly because of echocardiographic findings (n = 18/6%). An echocardiography-driven target vessel change was necessary in 33 pts. (11%). In the 312 pts. who received ethanol, permanent pacing was necessary in 22 cases (7%). In-hospital mortality was 1.3% (4 pts.). After 3 months, mean NYHA functional class was reduced from 2.9 +/- 0.5 to 1.5 +/- 0.6 (p < 0.0001) along with a gradient reduction from 60 +/- 33 to 13 +/- 18 mmHg at rest, and from 120 +/- 43 to 38 +/- 35 mmHg with provocation (p < 0.0001 each). Exercise capacity improved from 94 +/- 51 to 115 +/- 43 W, peak oxygen consumption from 18 +/- 4 to 21 +/- 6 ml/kg/min (p < 0.01 each). There was no significant difference regarding residual gradients in pts. with different levels of immediate gradient reduction during probatory balloon occlusion. CONCLUSIONS: Catheter-based septal ablation is an effective non-surgical technique for reducing symptoms and outflow gradients in HOCM. Intra-procedural echocardiographic guidance has a cumulative impact on the interventional strategy in about 15-20%, and clearly identifies pts. who should not receive ethanol but undergo a surgical myectomy. PMID- 15341871 TI - Biventricular diastolic behaviour in patients with hypertrophic and hereditary hemochromatosis cardiomyopathies. AB - AIM: To define biventricular diastolic behaviour in patients with cardiomyopathies with predominant diastolic left ventricular (LV) dysfunction. METHODS AND RESULTS: Doppler tissue echocardiography and both mitral and tricuspid Doppler inflow profiles were investigated in hypertrophic (n = 17), hereditary hemochromatosis (n = 12) cardiomyopathies and age-matched normals (n = 31). The cardiomyopathy group had both lower early diastolic mitral lateral annular (El), cm/s (13.9 +/- 6.5) and medial (Em) (10.0 +/- 4.5) velocities compared with normals (19.5 +/- 5.5, 15.9 +/- 3.4, p < 0.01, respectively). In the cardiomyopathy group, late isovolumic relaxation myocardial velocity gradient (IVR-MVG) (s(-1)) was positive compared with negative in normals (1.3 +/- 1.3 vs. -0.7 +/- 1.4, p < 0.01, respectively). In both the cardiomyopathy group and in normals the onset of the tricuspid E-wave preceded the onset of the mitral E wave. However, the onset of early diastolic tricuspid annular (Et) motion preceded the onset of El (ms) only in normals, but not in the cardiomyopathies (43 +/- 26 vs. -8 +/- 44, p < 0.01, respectively). In the cardiomyopathy group there was a positive correlation between the onset of Et and abnormally positive late IVR-MVG (r = 0.51, p = 0.002). CONCLUSIONS: Biventricular early diastolic behaviour is abnormal in the selected group of cardiomyopathy patients. The delay in the Et (early diastolic longitudinal right ventricular relaxation) may have a negative effect on LV diastolic function. PMID- 15341872 TI - Inter- and intra-study reproducibility of contrast echocardiography for assessment of interventricular septal wall perfusion rate in humans. AB - AIMS: To assess inter- and intra-study reproducibility of myocardial contrast echocardiography (MCE) imaging for quantitative measurement of myocardial perfusion rate in humans in ambulatory setting. METHODS AND RESULTS: In 20 subjects, we performed 2 MCE tests 20-40 min apart on the same day under the same standardized protocol, and evaluated single-reader between-study and between reading reproducibility of assessment of indicators of myocardial perfusion rate, such as the slope of video-intensity change k, and the factors A (peak video intensity) and B (background video-intensity after bubble destruction) and the product k X A. The region of interest was placed at the mid-posterior interventricular septal wall visualized in apical 4-chamber view. In a sub analysis, we evaluated indicators of myocardial perfusion rate comparing subjects with normal vs. those with subnormal ejection fraction (EF). Inter-study reproducibility of assessment of k was relatively low (intraclass correlation coefficient = 0.36), whereas intra-study reproducibility was fair (intraclass correlation coefficient = 0.61). The parameters k X A and B showed higher reproducibility than the k (inter- and intra-study intraclass correlation coefficients 0.64 and 0.75, 0.74 and 0.91, respectively). For reference, reproducibility data of the depth of the region of interest, of EF and CO were excellent. k and k X A were lower in subjects with low vs. those with normal EF. Only k and k X A were lower in subjects with subnormal than in those with normal EF. CONCLUSIONS: The MCE-derived indicator of myocardial perfusion rate k X A showed fairly good between-study and between-reading reproducibility. PMID- 15341873 TI - Tissue motion imaging of the left ventricle--quantification of myocardial strain, velocity, acceleration and displacement in a single image. AB - AIMS: Several methods of parametric imaging of left ventricular function including tissue velocity imaging (TVI) and strain rate imaging (SRI) have previously been presented, however, they have the limitation that they can, respectively, portray only one physiological myocardial parameter. The aims of this pilot study were to implement and validate tissue motion imaging (TMI) for the first time, a visualization technique which permits acceleration, velocity, displacement and strain to be interpreted quantitatively or semi-quantitatively in a single image. METHODS AND RESULTS: TMI is achieved by the color coding of temporal tissue velocity integrals. The principles behind this technique are validated, and case examples demonstrating its use in the clinical setting are provided. Limitations of the method as well as future applications and improvements are discussed. CONCLUSION: As this method allows representation of a multitude of variables and is visually attractive, it may facilitate more widespread use of myocardial quantitation in everyday practice. PMID- 15341874 TI - Reversal of cardiac abnormalities in a young man with idiopathic hypereosinophilic syndrome using a tyrosine kinase inhibitor. AB - We report a case of idiopathic hypereosinophilic syndrome in a young man, who showed symptoms and electrocardiographic findings mimicking an acute coronary syndrome. Two-dimensional echocardiography, together with laboratory data, allowed us to make the diagnosis and to start a treatment with imatinib mesylate, a 2-phenylamonopyrimidine-based tyrosine kinase inhibitor, which reversed the cardiac abnormalities. PMID- 15341875 TI - Aneurysm of the membranous septum with ventricular septal defect and infective endocarditis. PMID- 15341876 TI - Arrhythmogenic right ventricular dysplasia presenting as acute coronary syndrome: a case report. AB - Arrhythmogenic right ventricular dysplasia (ARVD) is underdiagnosed cardiomyopathy which commonly presents in young adults with ventricular tachycardia or sudden death. We report a case of ARVD presenting with features of acute coronary syndrome. The suspicion of ARVD came only when echocardiogram revealed abnormal shape and wall motion of right ventricle, which was later confirmed by right ventricular angiogram. The diagnosis of ARVD was discussed and the literature reviewed. PMID- 15341878 TI - Development of and clinical experience with capecitabine (Xeloda) in the treatment of solid tumours. AB - The oral fluoropyrimidine capecitabine (Xeloda) delivers 5-FU to the tumour site, thereby limiting the side effects and other complications associated with intravenous (i.v.) 5-FU. As an oral drug, capecitabine is preferred to 5-FU by many patients as it can be conveniently taken at home. In first-line metastatic colorectal cancer (MCRC), capecitabine results in superior response rates and equivalent progression-free and overall survival compared with i.v. 5-FU/LV. There is also increasing evidence for replacing i.v. 5-FU with capecitabine in combination with other anticancer agents (e.g. oxaliplatin and irinotecan) in MCRC and in the adjuvant treatment of early stage colon cancer. In anthracycline pretreated metastatic breast cancer (MBC), adding capecitabine to docetaxel improves survival, time to progression (TTP) and response rates beyond docetaxel. Single-agent capecitabine is also effective in pretreated MBC and is a promising first-line therapy. Capecitabine has a favourable safety profile, the most frequent adverse events being hand-foot syndrome, stomatitis and diarrhoea. Because capecitabine is orally administered, it is possible to intervene promptly with dose interruption/reduction to resolve adverse events without impacting on efficacy. The increasing availability of capecitabine in the home-based setting requires careful consideration of the role of the oncology nurse, who is the key link between the patient and clinician for effective and efficient management. PMID- 15341879 TI - Management of adverse events and other practical considerations in patients receiving capecitabine (Xeloda). AB - As capecitabine (Xeloda) is converted to 5-FU within tumours it can produce 5-FU like side effects. However, diarrhoea, stomatitis, nausea, alopecia and neutropenia are significantly less frequent than with i.v. 5-FU. Hand-foot syndrome (HFS) is the only clinical adverse event occurring more often during capecitabine treatment. These findings in MCRC have also been confirmed in a large phase III trial in early stage colon cancer (X-Act adjuvant study) and phase II clinical trials in metastatic breast cancer. Because capecitabine is taken in the outpatient setting, the nurse and/or supervising clinician are responsible for educating patients how to use it correctly and on the nature/recognition/severity of adverse events. Patients need to be aware that temporary interruptions/dose modifications do not reduce the overall efficacy of capecitabine and will most likely lead to a resolution of side effects. Consequently, oncology nurses will be assuming a more significant and pivotal role in the efficient education and support of patients during home-based therapy with capecitabine. PMID- 15341880 TI - Management of hand-foot syndrome in patients treated with capecitabine (Xeloda). AB - Comparative trials of capecitabine (Xeloda) versus 5-FU/LV in metastatic colorectal cancer have shown that hand-foot syndrome (HFS) was the only clinical adverse event occurring more frequently with capecitabine. Most patients with HFS present with dysesthesia, usually with a tingling sensation in the palms and soles of the hands and feet. This can progress in 3-4 days to burning pain plus well-defined symmetric swelling and erythema. The hands tend to be more commonly affected than the feet, and might even be the only area affected in some patients. HFS can interfere with the general activities of daily living, especially when blistering, moist desquamation, severe pain or ulceration occurs. While HFS is manageable, if ignored it can progress rapidly. However, dose interruption and reduction of capecitabine usually leads to a rapid reversal of signs and symptoms without long-term consequences. Nurses play a key role in educating patients how to recognise HFS, when to interrupt treatment and how to adjust the dose to maintain effective therapy with capecitabine over the long term. It is particularly important that patients and nurses are aware that dose interruption/reduction does not affect the overall antitumour efficacy of capecitabine. PMID- 15341881 TI - The vital role of education and information in patients receiving capecitabine (Xeloda). AB - Use of oral capecitabine (Xeloda) as home-based therapy leads to savings in healthcare resources and costs and oral chemotherapy is preferred by many cancer patients over intravenous (i.v.) 5-FU. The demands of patient management for oral therapy differ significantly from those of i.v. chemotherapy. Consequently, cancer programmes have begun to implement strategies to meet these unique demands, offer educational programmes for clinicians who prescribe capecitabine, and consider potential changes in staff responsibilities. To encourage patients to take an active part in their care, which is vital with home-based oral therapy, they must be properly informed about their treatment. Patient information needs to be specific for the cancer type and relevant for the country. Various tools have been developed, including prescription guides, diary cards and support kits, which are useful in reinforcing verbal discussions about the use of capecitabine and in helping patients to manage their treatment. Nurses have a vital role to play in encouraging the optimal use of capecitabine and prompt management of adverse events, thereby enabling the patient to achieve a better clinical outcome and maintain an improved quality of life in the home environment. PMID- 15341882 TI - Implementation of capecitabine (Xeloda) into a cancer centre: UK experience. AB - Switching patients from intravenous 5-FU/LV to oral capecitabine (Xeloda) for the treatment of metastatic colorectal cancer is associated with a reduction in the need for hospitalisations to manage 5-FU-associated delivery and complications, with resulting healthcare savings. However, implementing oral therapy with capecitabine within a cancer centre in the UK has required a considerable change in attitude within healthcare services. The resulting need for patients to take an active role in their treatment, and the co-ordination and monitoring of such a service at home has raised issues for chemotherapy services. To enhance patient involvement and compliance with medication a patient guide was developed to educate, and support individuals and enable them to understand the rationale for treatment and when to seek help. In addition, patients are encouraged to monitor and record symptoms in a diary. This change in service focus has required an investment in time educating and informing patients, community health workers and hospital practitioners. This change has been co-ordinated through the chemotherapy outpatient clinic. Effective communication between hospital and home has been important in implementing oral chemotherapy. While the initial challenge of monitoring and educating patients receiving capecitabine has been met, the Primary Care team and cancer centre need to continue to assess side effects and patient compliance in order to improve knowledge of capecitabine among healthcare professionals and ensure safe practice. PMID- 15341883 TI - Implications of capecitabine (Xeloda) for cancer nursing practice. AB - Home-based therapy with oral capecitabine (Xeloda) has a number of advantages over i.v. hospital-based chemotherapy regimens, including improvement in patients' quality of life, and medical resource/cost savings compared with 5 FU/LV in metastatic colorectal cancer. In addition, the ability of capecitabine to extend survival beyond docetaxel in patients with previously treated metastatic breast cancer means that capecitabine plus docetaxel is a very cost effective combination. Oncology nurses should prepare for the increased use of oral capecitabine as a single agent or in combination regimens in the outpatient setting. The use of this drug requires enhanced patient education skills, communication (e.g. telephone contact) and patient management on the part of the nurse. Oncology nurses will also need to accept a more significant and pivotal role in the clinical oncology team, not only as a point of contact between the patient and clinician, but also in documenting the benefits of capecitabine to ensure the effective management of patients receiving the drug. PMID- 15341885 TI - Restless legs syndrome: the most common and least diagnosed sleep disorder. PMID- 15341886 TI - An emerging interdisciplinary sleep medicine perspective on the high prevalence of co-morbid sleep-disordered breathing and insomnia. PMID- 15341887 TI - Epidemiology of restless legs syndrome as diagnosed in UK primary care. AB - BACKGROUND AND PURPOSE: Restless legs syndrome (RLS) is a sensorimotor disorder with a general-population prevalence of 5-10%. Although, data on the prevalence of clinically diagnosed RLS are limited, and there are none regarding incidence, a shortfall compared with general-population values is likely, as not all patients are driven to consult. There may also be poor awareness of the condition among primary care physicians (PCPs). PATIENTS AND METHODS: The General Practice Research Database was used to gather prevalence and incidence data from UK PCPs and to investigate PCP awareness over the period 1994-1998. A total of 1,561,692 persons were covered by the database on January 1, 1999. RESULTS: A diagnosis of RLS was registered in 3877 patients, giving a prevalence of 0.25%. Incidence values were also low (41.0 per 100,000 person-years), increased with age and were higher in women than in men. Although, at least one RLS diagnosis was made in 94% of medium-to-large practices, sufferers were more likely than age/sex-matched controls to be diagnosed with conditions commonly confused with RLS in 2 years before receiving their RLS diagnosis. Furthermore, at the time of diagnosis, many sufferers were prescribed medications not effective in RLS (principally, oxerutins and quinine). CONCLUSION: These data are largely consistent with a lack of awareness of RLS among PCPs in the UK in the period studied. PMID- 15341888 TI - The impact of extended sleep on daytime alertness, vigilance, and mood. AB - BACKGROUND AND PURPOSE: To measure the effects of prolonged sleep extension on daytime alertness, vigilance, and mood in healthy young adults. Little research has documented the effects of increased sleep on daytime function despite a high prevalence of daytime fatigue and sleepiness in the adult population. Past extension studies report conflicting results with regard to Multiple Sleep Latency Test (MSLT) scores, vigilance, and mood ratings. No study has challenged subjects to maximum sleep extension, defined by an MSLT score of 20. PATIENTS AND METHODS: Fifteen healthy college students reporting minimal daytime sleepiness were allowed to sleep as much as possible during a sleep extension period. MSLT scores, psychomotor vigilance task (PVT) reaction times, and profile of mood states (POMS) ratings were measured at baseline, mid-extension, and end extension. RESULTS: There was a significant increase in both journal and actigraphy sleep totals during all extension segments (P<0.01). MSLT scores increased significantly from baseline to both mid- and end-extension (P<0.01). Five of eight tabulated PVT measures also improved significantly at mid- and end extension with respect to baseline (P<0.05). POMS vigor and fatigue scores showed a similar improvement (P<0.01). Seven subjects achieved an MSLT score of 20. Six subjects showed substantial improvements while two subjects obtained relatively little extra sleep and showed little or no MSLT improvement. The maximum extension group displayed exceptional improvements in vigilance and POMS ratings. CONCLUSIONS: Extended sleep leads to substantial improvements in daytime alertness, reaction time, and mood. PMID- 15341889 TI - Frequency of insomnia report in patients with obstructive sleep apnoea hypopnea syndrome (OSAHS). AB - BACKGROUND AND PURPOSE: Insomnia and Obstructive Sleep Apnoea Hypopnea Syndrome (OSAHS) are the two most common sleep disorders, and both have significant associated health costs. Despite this, relatively little is known about the prevalence or impact of insomnia in those with OSAHS, although a recent study suggested there may be substantial comorbidity between these disorders [Chest 120 (2001) 1923-9]. The primary aim of this study was to further explore the prevalence of insomnia in OSAHS. A secondary aim was to assess the effect of factors that may impact on both conditions, including mood and sleep-beliefs. PATIENTS AND METHODS: Consecutive patients referred to an accredited Sleep Investigations Unit [n = 105] completed a brief standardized battery of validated questionnaires assessing sleep-related variables and mood. RESULTS: Results showed a high rate of prevalence of clinical insomnia in this OSAHS population, and a strong positive correlation between OSAHS and insomnia symptom severity. Further, OSAHS patients with comorbid insomnia had increased levels of depression, anxiety and stress compared to patients with OSAHS-only, and both patient groups reported similar and significant levels of dysfunctional beliefs about sleep. Findings in relation to habitual sleep, assessed using subjective (diary) and objective criteria (polysomnogram), were mixed but generally showed greater sleep disturbance among those with OSAHS-insomnia compared to those with OSAHS-only. CONCLUSIONS: Overall these findings suggest that comorbidity of insomnia in OSAHS patients may lead to increased OSAHS severity and that patients with both conditions may experience more symptoms relating to depression, anxiety and stress. These findings underscore the need for insomnia assessment and management services, even in clinics that primarily service patients with OSAHS. PMID- 15341890 TI - Correlates of benzodiazepine use in individuals with insomnia. AB - BACKGROUND AND PURPOSE: Although benzodiazepines (BZDs) are commonly used in the treatment of insomnia, there is little information about psychological, health, and sociodemographic correlates associated with their use. OBJECTIVE: This study examined correlates of benzodiazepine use for sleep in a clinical sample of patients seeking treatment for insomnia at a sleep disorders clinic. PATIENTS AND METHODS: The sample consisted of 97 individuals evaluated at a sleep disorders clinic for a presenting complaint of insomnia. Two groups were formed, including one with 61 patients who had been using BZDs for sleep for an average of 56.6 months (SD=68.0), and another with 36 patients who had insomnia but were not using BZDs or any other sleep aid. Logistic regressions, adjusting for an age difference between the user and non-user groups, were performed to examine variables associated with BZD use. Independent variables included demographic, medical, insomnia-related and psychological parameters and subjective sleep-wake characteristics. Unadjusted regression analyses were performed to identify factors associated with a pattern of long-term use (>12 months) within the user group. RESULTS: Age was a significant predictor of BZD use (OR=1.84, P = 0.0002). Significant age-adjusted predictors of BZD use included perceived insomnia severity (OR=1.17, P = 0.038), depressive symptoms, (OR=1.108, P = 0.009) and state and trait anxiety symptoms (OR=1.062, P = 0.016; OR=1.084, P = 0.005, respectively). Significant predictors of long-term use (>12 months) were age of insomnia onset (OR=0.951, P = 0.0214), more frequent BZD use (OR=3.284, P = 0.0221), and higher state-anxiety (OR=1.106, P = 0.0471). CONCLUSIONS: Age, psychological variables and perceived sleep disturbances severity, are associated with BZD use in patients with insomnia. PMID- 15341891 TI - 'Hypnotic' prescription patterns in a large managed-care population. AB - BACKGROUND AND PURPOSE: Medical treatment of insomnia has declined over the past decade and, when treated, use of non-hypnotic medications has increased. This study assessed the characteristics of the prescriptions for insomnia and of the patients receiving those prescriptions. PATIENTS AND METHODS: The outpatient pharmacy database of the Henry Ford Hospital, Health Alliance Plan (HAP) was searched from 1/1/98 to 6/30/99 for mentions of the 10 most frequently used drugs for the treatment of insomnia listed in the National Disease and Therapeutic Index for 1987-1996. The 10 drugs were alprazolam, amitriptyline, clonazepam, doxepin, flurazepam, lorazepam, temazepam, trazodone, triazolam, and zolpidem. These were classified by their indication as antidepressant, anxiolytic, or hypnotic and the three indication groupings were compared on patient and prescription characteristics. RESULTS: Over the 18 month period the total patient population covered by HAP was 287,456; 20,014 (7%) patients received one or more prescriptions for insomnia. Of these, anxiolytics were most frequently prescribed (55%), then antidepressants (25%), and hypnotics least frequently (20%). Patients receiving hypnotics were more likely to be male, older, and to receive a narrower dose range, in smaller quantities and with fewer refills than patients receiving anxiolytics or antidepressants. CONCLUSIONS: In this large managed-care population, hypnotics are prescribed conservatively, while the non-hypnotics, for which there is limited efficacy and safety data, are prescribed on a more chronic basis. PMID- 15341892 TI - Health-related quality of life in Italian patients with narcolepsy: the SF-36 health survey. AB - OBJECTIVE: To evaluate the psychometric performances of the 36-item short form (SF-36), a tool measuring health-related quality of life (HRQoL) in patients with narcolepsy or other sleep disorders, presenting excessive daytime sleepiness (EDS) as the main symptom. To compare the HRQoL of patients with narcolepsy with the Italian norm. To disclose the clinical predictors influencing the HRQoL. PATIENTS AND METHODS: One hundred and eight adults with narcolepsy, 115 with obstructive sleep apnea syndrome (OSAS) and 42 with idiopathic hypersomnia (IH) enrolled for a multicentric cross-section study employing self-administered questionnaires investigating the HRQoL (SF-36), EDS and cataplexy. A psychometric analysis and the standardized scoring of the 8 scales of the SF-36 were performed. The correlation between SF-36 scales and clinical features of narcolepsy was measured by a multiple linear regression analysis. RESULTS: SF-36 satisfied the recommended psychometric standards. Patients with narcolepsy had all domains except Bodily Pain lower than the Italian norm. Some of the variance of Physical Functioning (R(2) 0.07), Role functioning-Physical (R(2) 0.31), General Health (R(2) 0.15), Vitality (R(2) 0.10), Social Functioning (R(2) 0.21) and Role functioning-Emotional (R(2) 0.15) was explained both by EDS (inverse correlation) and disease duration (direct correlation). CONCLUSIONS: The SF-36 form is a reliable outcome measure for hypersomniac disorders. Narcolepsy seriously impacts patients' HRQoL in terms of physical and mental well-being and impairs physical and emotional functions, restricting social life. EDS is the main symptom determining this impairment. However, disease duration plays a role in counterbalancing this feature, probably by means of adaptation. PMID- 15341893 TI - Functional status in patients with narcolepsy. AB - OBJECTIVE: To describe health and functional status in treated narcolepsy patients and to compare it with normative data and with patients with another cause of sleepiness, the obstructive sleep apnea/hypopnea syndrome (OSAHS) patients. METHODS: A functional status survey in narcolepsy patients and in symptomatic untreated and CPAP treated OSAHS patients with an apnea-hypopnea index (AHI) >5 was assessed using the UK Short Form 36 (SF-36) questionnaire, the Functional Outcomes of Sleep Questionnaire (FOSQ) and the Epworth Sleepiness Scale (ESS). SF-36 scores in narcoleptics were compared to age and sex matched controls. Narcolepsy patients also replied to a psychosocial aspects questionnaire. Health history and demographic data were obtained via a review of medical records and postal survey. RESULTS: Forty-nine treated narcolepsy patients, 56 untreated OSAHS and 48 CPAP treated OSAHS patients attending the sleep disorders clinic were recruited for this study. Narcoleptics presented significantly lower scores in all SF-36 domains compared to normative data. No difference in SF-36 scores was found between narcoleptics and untreated OSAHS patients but narcoleptics were sleepier and had lower FOSQ scores. These treated narcolepsy patients had lower scores in two dimensions of the SF-36 and in all FOSQ domains compared to CPAP-treated OSAHS patients. CONCLUSIONS: Functional status in treated narcoleptics is poor. PMID- 15341894 TI - Modeling the causal relationships between symptoms associated with restless legs syndrome and the patient-reported impact of RLS. AB - The objective of this study is to examine the causal relationships between the symptoms of restless legs syndrome (RLS) and specific clinical and subjective health-related, quality of life consequences. Structural equation modeling was applied to data from a questionnaire-based observational study. The RLS morbidities of decreased functional alertness and emotional distress in our sample of patients appear to be mostly secondary to the sleep disturbance associated with RLS. There was no clear indication of any other feature of RLS affecting these two aspects of RLS morbidity. A primary treatment goal should be the reduction of the sleep disturbance of RLS, both to decrease the RLS-related nocturnal distress and to improve daytime functioning. PMID- 15341895 TI - Evening and morning blood gases in patients with obstructive sleep apnea. AB - BACKGROUND AND PURPOSE: The purpose of this study was to see if blood oxygen levels deteriorate overnight during obstructive sleep apnea (OSA). Before and after sleep, arterial blood gases (ABGs) in OSA subjects and controls were drawn during a diagnostic night, as well as during a continuous positive airway pressure (CPAP) night for the OSA subjects. PATIENTS AND METHODS: Subjects, both male and female, were referred to our sleep laboratory for symptoms of daytime somnolence. Subjects consisted of a control group (N=13) with a mean apnea hypopnea index (AHI) of 3.3 events/h and a study group (N=22) with a mean baseline AHI of 57 events/h. RESULTS: With the subject supine, resting room air ABGs were drawn at 'lights out' on the evening before (PM) nocturnal polysomnography and in the morning (AM) at discontinuation ('lights on') of the sleep study. In controls, PM PaO(2) (79.4+/-9.7 mmHg) was not significantly different from AM PaO(2) (80.2+/-8.9 mmHg, P=0.5). In apneic subjects, the PM PaO(2) was 78.7+/-7.2 mmHg compared to an AM PaO(2) of 72.6+/-8.3 mmHg (P<0.05). The AM PaO(2) after a night of CPAP treatment in the OSA subjects was 77.5+/-10.2 mmHg compared to the PM PaO(2) of 76.0+/-6.0 mmHg (NS). The PM and AM PaCO(2)s were not different in controls or in study subjects under baseline conditions. However, during titration with nasal CPAP, the PaCO(2) was significantly higher in the morning after CPAP treatment [43.1+/-4.8 vs. 46.1+/-4.8 mmHg, respectively (P<0.05)]. CONCLUSIONS: OSA subjects showed a fall in overnight resting oxygenation. This could be accounted for by overnight deterioration of gas exchange and is ameliorated by CPAP. PMID- 15341896 TI - Restless legs syndrome: clinical experience with long-term treatment. AB - BACKGROUND AND PURPOSE: There are limited data on long-term treatment efficacy, and almost none on predictors of treatment response in patients with restless legs syndrome (RLS). To assess: (1) long-term efficacy of RLS treatment in a clinical setting, (2) predictors of a good treatment response, and (3) the value of the RLS-severity score according to the criteria of the International Restless Legs Syndrome Study Group (IRLSSG). PATIENTS AND METHODS: Over three years 70 patients (36 men, 34 women; mean age: 59 years; range: 29-79) with RLS were prospectively assessed. Diagnosis of RLS was made according to international criteria Severity of RLS symptoms was were assessed at the outset by the IRLSSG rating scale. Treatment was chosen individually according to clinical judgement. After a mean follow-up time of 16 months (range: 1-106 months) evolution of symptoms was assessed by both overall clinical impression and IRLSSG rating scale. Clinical characteristics and treatment effect were compared between patients never treated for RLS before this study ('naive'=N-pts) and those with previous treatment ('treated'=T-pts). Predictors of treatment response were sought for comparing patients with good treatment response (good, better or much better on follow-up) and those with bad (B-pts) treatment response. RESULTS: There were 40 N-pts and 30 T-pts. The mean IRLSSG score (hereinafter, IRLSSG) at baseline was 26 (range 12-38). No significant differences were found between N and T-pts in age, gender, etiology and duration of RLS, positive family history, presenting sleep complaint, IRLSSG, or percentage of patients with periodic limb movements in sleep (PLMS) on polysomnography (PSG). At final follow-up 30 (76%) of 40 N-pts and 23 (77%) of 30 T-pts had a good (G-pts) treatment response. The mean IRLSSG at follow-up was 19 (range:1-36). There was a significant correlation between improvement of overall clinical impression (better or much better on final follow-up) and reduction of IRLSSG (P<0.0001). PLMS were more common in B- than G-pts (100 vs 58% of patients, P=0.02). In all other variables considered the two groups were similar. CONCLUSION: (1) A good long-term treatment response can be obtained and maintained in a clinical setting in about 80% of RLS patients. (2) Patients with RLS and without PLMS may have a better long-term treatment response, and (3) the IRLSSG is a useful tool for assessment of evolution of RLS symptoms over time in individual patients. PMID- 15341897 TI - Occult sleep apnea: the dilemma of negative polysomnography in symptomatic patients. AB - BACKGROUND AND PURPOSE: To determine the benefit of repeat polysomnography with/without esophageal pressure (PES) monitoring to diagnose sleep apnea syndrome (SAS) in patients with symptoms of sleep apnea who have had a 'negative', single-night polysomnogram (PSG). PATIENTS AND METHODS: This is a retrospective investigation of 1187 patients seen in our sleep lab from January to December 2001, of which 709 were adults suspected of having sleep apnea. Following a single PSG, 588 patients were diagnosed with sleep apnea and 121 had negative PSGs (an apnea-hypopnea index <5 events per hour). Of the 121 patients, 92 continued to complain of unexplained sleepiness, loud snoring, or apnea, symptoms which were also documented on their initial evaluation (PSG or multiple sleep latency testing). The remaining 29 patients had no further complaints, or another medical cause of their sleepiness was established (i.e. asthma) following the single-night PSG. Of the 92 patients, 28 underwent additional screening with both repeat PSG and PES monitoring within the following 6 months. RESULTS: With repeat PSG and PES monitoring, 18 of the 28 patients with previous, negative PSGs were diagnosed with sleep apnea. The sensitivity of a single-night PSG fell to 97%, with a false negative rate of 3%. Only 12 of the 28 would have been positive based on polysomnographic criteria alone, without the additional PES monitoring. On the other hand, 10 of the 28 remained negative and further evaluation revealed other, underlying medical problems (i.e. nocturnal asthma) that explained their symptoms. CONCLUSIONS: There is a clear benefit of repeat PSG, with or without PES monitoring, for patients with a prior negative PSG and continued symptoms suspected of having SAS. PMID- 15341898 TI - Effectiveness of mirtazapine in the treatment of sleep apnea/hypopnea syndrome (SAHS). AB - Several drugs have been described as possible treatments for Sleep Apnea/Hypopnea Syndrome (SAHS) but the data available does not support their use. In an animal model of central apnea the use of mirtazapine produced a significant reduction of apneas. We present a male patient, 82 years old, with excessive daytime sleepiness and loud snoring during at least 10 years. An overnight polysomnography (PSG) revealed an apnea/hypopnea index of 54.9 events per hour of sleep with a minimum pulse oximetric saturation (SaO(2)) of 78% and an arousal index of 40.4 per hour. A nasal CPAP titration in the second half of the night showed suppression of apneas with a CPAP level of 8 cmH(2)O. The patient refused to use the CPAP device and began with 15 mg of mirtazapine at bedtime. A second PSG performed after 3 months of mirtazapine showed a significant reduction in the apnea/hypopnea index (9.3 events per hour of sleep; 81% minimal oxygen saturation (SaO(2))). Clinically, the patient and his wife reported a clear reduction of excessive daytime sleepiness and an improvement in self-reported functioning and well-being without any important side effects. This successful case appears to be the first report with mirtazapine in human SAHS and supports the need for an appropriate clinical trial with this drug. PMID- 15341899 TI - Concomitant use of modafinil and tranylcypromine in a patient with narcolepsy: a case report. AB - This is a case report of a patient with narcolepsy treated simultaneously with modafinil and tranylcypromine. The concurrent use of these two medications is normally contraindicated based on theoretical concerns. To our knowledge this is the first report of the combined use of these two medications in a patient for any reason. This combination of medications was particularly effective in the treatment of the patient's narcoleptic symptoms and was well tolerated, with no evidence of the potential cardiovascular side effects. Due to modafinil's lack of sympathomimetic activity and unique pharmacological profile, it may be well tolerated with monoamine oxidase inhibitor's (MAOI's) in humans with respect to hemodynamic parameters and concerns of hypertensive crisis. Further investigation would help clarify cardiovascular safety issues in using this combined therapy. We believe that with proper informed consent from the patient and careful monitoring, a combination of an MAOI and modafinil may be a viable treatment alternative in refractory cases of narcolepsy. PMID- 15341900 TI - What's wrong with sleep hygiene? PMID- 15341901 TI - Oxidative DNA damage and disease: induction, repair and significance. AB - The generation of reactive oxygen species may be both beneficial to cells, performing a function in inter- and intracellular signalling, and detrimental, modifying cellular biomolecules, accumulation of which has been associated with numerous diseases. Of the molecules subject to oxidative modification, DNA has received the greatest attention, with biomarkers of exposure and effect closest to validation. Despite nearly a quarter of a century of study, and a large number of base- and sugar-derived DNA lesions having been identified, the majority of studies have focussed upon the guanine modification, 7,8-dihydro-8-oxo-2' deoxyguanosine (8-OH-dG). For the most part, the biological significance of other lesions has not, as yet, been investigated. In contrast, the description and characterisation of enzyme systems responsible for repairing oxidative DNA base damage is growing rapidly, being the subject of intense study. However, there remain notable gaps in our knowledge of which repair proteins remove which lesions, plus, as more lesions identified, new processes/substrates need to be determined. There are many reports describing elevated levels of oxidatively modified DNA lesions, in various biological matrices, in a plethora of diseases; however, for the majority of these the association could merely be coincidental, and more detailed studies are required. Nevertheless, even based simply upon reports of studies investigating the potential role of 8-OH-dG in disease, the weight of evidence strongly suggests a link between such damage and the pathogenesis of disease. However, exact roles remain to be elucidated. PMID- 15341902 TI - Importance of allele frequency estimates in epidemiological studies. AB - This study addresses the issue of appropriate allelic frequency estimates in epidemiological studies. Reasons for imprecise estimate of allele frequency may be population stratification, and lack of power of many published studies to define true allele frequencies in the general population. As an example of the lack of power of epidemiological studies, we plot the frequency of GSTM1 deletion versus sample size for the 79 studies from the GSEC pooled analysis. The estimate of allele frequency derived from small groups of controls deviates more from the true frequency than the estimate derived from larger studies. We discuss the possible consequences of not properly defining allele frequencies in the population. This may reflect on the conduct of association studies, on assessment of the effects of multigenic mechanisms, and on the determination of genetic diversity. PMID- 15341903 TI - Know-how of RNA interference and its applications in research and therapy. AB - Double stranded RNA (dsRNA) mediates gene silencing in a sequence specific manner. Originally recognized in plants and lower organisms, it was recently extended to higher eukaryotes and established as an important evolutionary conserved phenomenon. It has been established that the double stranded short interfering RNAs (siRNAs) originate by the activity of a dsRNA-specific endonuclease, Dicer. siRNA in conjunction with a multiple enzyme complex called RNA-induced silencing complex (RISC) locates to the specific sites on mRNA and degrades it by endonuclease and exonuclease activities. In addition to gene silencing at transcript level (degradation of messenger RNA), siRNA was also shown to reduce the expression of protein by silencing of gene promoters via de novo methylation. By virtue of their specific gene silencing activity and owing to the recent discoveries on their plasmid and virus driven expression, small dsRNAs are being widely adopted in research and therapeutics. They are rapidly replacing the conventional gene knock-out technologies. siRNA libraries are also being recruited as a new tool in genome wide functional screenings. There is no doubt that further progress in understanding the mechanism of their action as well as strategies to achieve their tightly regulated and tissue specific expression will revolutionize basic and applied biomedical research. PMID- 15341904 TI - Telomere dysfunction in genome instability syndromes. AB - Telomeres are nucleoprotein complexes located at the end of eukaryotic chromosomes. They have essential roles in preventing terminal fusions, protecting chromosome ends from degradation, and in chromosome positioning in the nucleus. These terminal structures consist of a tandemly repeated DNA sequence (TTAGGG in vertebrates) that varies in length from 5 to 15 kb in humans. Several proteins are attached to this telomeric DNA, some of which are also involved in different DNA damage response pathways, including Ku80, Mre11, NBS and BLM, among others. Mutations in the genes encoding these proteins cause a number of rare genetic syndromes characterized by chromosome and/or genetic instability and cancer predisposition. Deletions or mutations in any of these genes may also cause a telomere defect resulting in accelerated telomere shortening, lack of end-capping function, and/or end-to-end chromosome fusions. This telomere phenotype is also known to promote chromosomal instability and carcinogenesis. Therefore, it is essential to understand the interplay between telomere biology and genome stability. This review is focused in the dual role of chromosome fragility proteins in telomere maintenance. PMID- 15341905 TI - What is acute disseminated encephalomyelitis (ADEM)? AB - Acute demyelinating encephalomyelitis (ADEM) is increasingly recognized but difficult to define precisely: eleven papers have recently been published that used different definitions. Several diseases, most of them relapsing, can mimic ADEM at onset and require specific treatments. First, the distinction between early signs of multiple sclerosis and ADEM is controversial in the absence of specific biological characterization. To clarify it, epidemiological evaluations on large groups of children, of calculated risk factors for the occurrence of relapses after a first demyelinating event are probably the best approach. Macrophage activation syndrome and different types of angiitis may mimic ADEM, because they can occur in the absence of the usual non neurological symptoms. Both entities require specific treatments. PMID- 15341906 TI - Aicardi-Goutieres syndrome: neuroradiological findings after nine years of follow up. AB - The main radiological features of Aicardi-Goutieres syndrome include basal ganglia calcification, cerebral atrophy and white matter alterations. We present a case where progress of cerebral calcifications demonstrated on consecutive CT scans later on was followed by a decrease. The MRI showed a progressive and significant loss of white matter and severe signal changes of the remaining myelin. The aetiology of the myelin changes and the transient worsening of the cerebral calcifications remains to be elucidated. It has previously been shown that the spinal fluid level of interferon-alpha decreases with age and we suggest that the biphasic course of the calcifications and the ventricular size as well as the clinical course shown in many patients with Aicardi-Goutieres might favor a causal role of interferon-alpha in the disease leading to a transient microangiopathy. PMID- 15341907 TI - Spontaneous arachnoid cyst rupture in a previously asymptomatic child: a case report. AB - Arachnoid cysts are benign congenital cavities arising in the subarachnoid space. Non-traumatic subdural effusion of cerebrospinal fluid is a rare complication requiring surgical treatment. CASE REPORT: A 15-year-old boy was admitted to the hospital because of symptoms related to acute intracranial hypertension (headache and vomiting). The cerebral CT-scan revealed a subdural hygroma adjoining a voluminous Sylvian arachnoid cyst. Two arachnoid cysts were incidentally discovered 11 years before this dramatic complication. Moreover, the patient had suffered a cerebral concussion 2 years earlier, but interestingly did not develop cystic hemorrhage or rupture, contrary to numerous cases previously described in the literature. The location of the cysts and their regular follow-up did not allow foreseeing a cystic rupture. Hygroma evacuation was first performed after which a subdural peritoneal shunting, using a programmable opening pressure valve, was implanted. Spontaneous rupture into the subdural space represents an unusual complication of arachnoid cysts. Clinical aspects, radiographic findings, pathogenesis and surgical management are described. It is important to point out that subdural hygroma or haematoma should never be excluded in the absence of trauma history, even in the case of small non-progressive cysts regularly supervised. PMID- 15341908 TI - Peripheral neuropathy--a rare complication of paediatric systemic lupus erythematosus. AB - Peripheral neuropathy is an uncommon complication in paediatric systemic lupus erythematosus (SLE). We report the case of a 10-year-old Chinese girl who developed peripheral neuropathy within 3 months of the onset of SLE and presented with bilateral foot drop and sensory symptoms of both hands and feet. There was no involvement of the central nervous system at the time of presentation. The patient was negative for anticardiolipin antibodies, but positive for lupus anticoagulant. She was treated with intravenous methylprednisolone followed by oral steroids, methotrexate, gabapentin and amitryptyline. Although peripheral neuropathy is a rare complication of paediatric systemic lupus erythematosus, one should be vigilant for this entity as part of the neurological spectrum. It may not be associated with involvement of the central nervous system. Antiphospholipid antibodies may have role in the pathogenesis of SLE associated peripheral neuropathy. We speculate that routine nerve conduction studies may have a role in detecting sub-clinical cases. PMID- 15341909 TI - Encephalopathy associated with influenza A. AB - We report a 3-year-old boy with influenza A virus-associated encephalopathy. The diagnosis was based on clinical findings, magnetic resonance imaging, repeated isolation of influenza A virus from nasopharyngeal aspirate and seroconversion between acute and convalescent sera. The clinical picture was characterized by fever and aphasia followed by seizures and a rapid deterioriation of consciousness 2 days later. This patient illustrates that influenza A-associated encephalopathy with severe neurological deficit can occur with atypical distribution of brain lesions during the course of the illness, while initial brain MRI and laboratory findings of cerebrospinal fluid are normal, including reverse transcription polymerase chain reaction. PMID- 15341910 TI - Succinic semialdehyde dehydrogenase deficiency (SSADH) (4-hydroxybutyric aciduria, gamma-hydroxybutyric aciduria). AB - Succinic semialdehyde dehydrogenase deficiency is one of the disorders of GABA metabolism, so it is not surprising that seizures occur as one of the symptoms in affected patients. Other features that are described include delayed development, hypotonia, myopathy with ragged red fibres, abnormal behaviour, and ocular abnormalities. Neonatal problems include prematurity, respiratory difficulties, and hypoglycaemia. The responsible gene has been identified on the short arm of chromosome 6. There are many mutations, and there is poor genotype-phenotype correlation resulting in difficulties in diagnosis. The pathogenesis of the condition is discussed, especially the results of the disturbed GABA catabolism, and the production of the gamma-hydroxybutyric acid. The many properties of this substance suggest it may act as a neurotransmitter or neuromodulator in the brain. The diagnosis may be difficult as the clinical picture is not really suggestive, but the MRI examination can help if it shows abnormalities in the globus pallidus. It will be confirmed by finding an excess of 4-hydroxybutyric acid in the body fluids; and the methods of estimation are discussed. Prenatal diagnosis is possible using a combination of methods. Treatment possibilities are limited. Vigabatrin should be of value as it is an inhibitor of GABA transaminase, but results have been disappointing. Symptomatic treatment may well be needed for control of seizures, abnormal behaviour and other disorders; and special educational needs must be served. PMID- 15341912 TI - GFP as a tool to analyze the organization, dynamics and function of nuclei and microtubules in Neurospora crassa. AB - We report the construction of a versatile GFP expression plasmid and demonstrate its utility in Neurospora crassa. To visualize nuclei and microtubules, we generated carboxy-terminal fusions of sgfp to Neurospora histone H1 (hH1) and beta-tubulin (Bml). Strong expression of GFP fusion proteins was achieved with the inducible Neurospora ccg-1 promoter. Nuclear and microtubule organization and dynamics were observed in live vegetative hyphae, developing asci, and ascospores by conventional and confocal laser scanning fluorescence microscopy. Observations of GFP fusion proteins in live cells largely confirmed previous results obtained by examination of fixed cells with various microscopic techniques. H1-GFP revealed dynamic nuclear shapes. Microtubules were mostly aligned parallel to the growth axis in apical compartments but more randomly arranged in sub-apical compartments. Time-lapse imaging of beta-tubulin-GFP in germinating macroconidia revealed polymerization and depolymerization of microtubules. In heterozygous crosses, H1-GFP and beta-tubulin-GFP expression was silenced, presumably by meiotic silencing. H1-GFP was translated in the vicinity of hH1+-sgfp+ nuclei in the common cytoplasm of giant Banana ascospores, but it diffused into all nuclei, another illustration of the utility of GFP fusion proteins. PMID- 15341913 TI - aflT, a MFS transporter-encoding gene located in the aflatoxin gene cluster, does not have a significant role in aflatoxin secretion. AB - The aflT gene resides between the polyketide synthase gene pksA and the P450 encoding cypA gene in the aflatoxin gene cluster of Aspergillus parasiticus. It is a single copy gene in the genome of A. parasiticus SRRC 2043 and SU-1 and was also found at the same relative position in the genome of Aspergillus flavus isolates. The predicted AFLT protein contained 14 transmembrane domains and had various degrees of the amino acid identity (34-56%) to fungal transporters belonging to the major facilitator superfamily. Targeted deletion of aflT in A. parasiticus SU-1 yielded transformants that were morphologically similar to SU-1. These aflT-deleted mutants produced and secreted aflatoxins comparable to the parental strain although they lost the production of the aflT transcript. Real time RT-PCR analysis showed that the expression of aflT was controlled neither by the aflatoxin pathway-specific activator AFLR nor by the co-activator AFLJ, which differed from the regulation of the aflatoxin biosynthetic genes pksA, nor1, ver1, and omtA. The FadA-dependent G-protein signaling pathway previously shown to govern aflatoxin biosynthesis and sporulation plays a role in the regulation of aflT expression. PMID- 15341914 TI - A retrotransposon-like element and its occurrence in British Columbia populations of Chondrostereum purpureum. AB - Repetitive DNA elements discovered in the basidiomycete Chondrostereum purpureum were characterized and validated for use as genetic markers. Regions of these marker sequences were similar to retrotransposon and retrotransposon-like sequences, as indicated by BLAST searches of NCBI databases. These sequences occur in multiple DNA fragments of variable length in a given C. purpureum isolate, and thus can serve as strain-specific genetic markers. The segregation of the markers within a progeny set demonstrated their stability through meiosis. The population structure of C. purpureum was assessed using the markers. There was no evidence of a barrier to gene flow between C. purpureum populations separated by 1400 km and no indication of population sub-structuring based on host or geographical source of isolate. Repetitive fragments were amplified from four other species, suggesting the occurrence of these retrotransposon-like elements in other basidiomycetes and the potential utility of these markers for other fungi. PMID- 15341915 TI - The analysis of the complete mitochondrial genome of Lecanicillium muscarium (synonym Verticillium lecanii) suggests a minimum common gene organization in mtDNAs of Sordariomycetes: phylogenetic implications. AB - The mitochondrial genome (mtDNA) of the entomopathogenic fungus Lecanicillium muscarium (synonym Verticillium lecanii) with a total size of 24,499-bp has been analyzed. So far, it is the smallest known mitochondrial genome among Pezizomycotina, with an extremely compact gene organization and only one group-I intron in its large ribosomal RNA (rnl) gene. It contains the 14 typical genes coding for proteins related to oxidative phosphorylation, the two rRNA genes, one intronic ORF coding for a possible ribosomal protein (rps), and a set of 25 tRNA genes which recognize codons for all amino acids, except alanine and cysteine. All genes are transcribed from the same DNA strand. Gene order comparison with all available complete fungal mtDNAs-representatives of all four Phyla are included-revealed some characteristic common features like uninterrupted gene pairs, overlapping genes, and extremely variable intergenic regions, that can all be exploited for the study of fungal mitochondrial genomes. Moreover, a minimum common mtDNA gene order could be detected, in two units, for all known Sordariomycetes namely nad1-nad4-atp8-atp6 and rns-cox3-rnl, which can be extended in Hypocreales, to nad4L-nad5-cob-cox1-nad1-nad4-atp8-atp6 and rns-cox3 rnl nad2-nad3, respectively. Phylogenetic analysis of all fungal mtDNA essential protein-coding genes as one unit, clearly demonstrated the superiority of small genome (mtDNA) over single gene comparisons. PMID- 15341916 TI - Analysis of heterologous expression of Candida albicans SEC61 gene reveals differences in Sec61p homologues related to species-specific functionality. AB - The protein secretory pathway has not been studied in depth in Candida albicans despite its essential role in the secretion of enzymes and cell surface components related to the ability of the fungus to colonize the human host. To gain further insight into the elements that participate in the first stages of the secretory process in this fungal pathogen we have isolated and characterized the C. albicans ortholog of SEC61. In other species SEC61 has been shown to encode the core element of the protein translocation apparatus within the ER membrane. The cloned gene appears to be essential for cell viability and encodes a highly conserved protein, very similar to the Sec61p from other yeast species both in sequence and hydropathy profile. However, CaSec61p is not able to complement the thermosensitive-growth phenotype of a Saccharomyces cerevisiae sec61 mutant, even though it is expressed and correctly incorporated into the ER membrane of the transformant cells. We report results indicating that the lack of functional complementation could be related to differences in the primary structure of the cytosolic domain located between the fourth and fifth transmembrane domains of the accepted topological model of Sec61p. PMID- 15341917 TI - Structural close-related aromatic compounds have different effects on laccase activity and on lcc gene expression in the ligninolytic fungus Trametes sp. I-62. AB - Nine phenolic compounds (p-coumaric acid, ferulic acid, guaiacol, syringol, p methoxyphenol, pyrocatechol, phloroglucinol, 3,5-dihydroxybenzoic acid, and syringaldazine) were tested for their ability to increase laccase production in the ligninolytic basidiomycete Trametes sp. I-62. All these compounds resulted in increases in laccase activity, with the highest levels being detected in the presence of p-coumaric acid (273-fold) and guaiacol (73-fold). The three laccase isozyme genes in this fungus lcc1, lcc2, and lcc3 are differentially expressed in the presence of some of these aromatics with total lcc transcript levels differing markedly depending on the aromatic compound tested. Guaiacol (the best inducer of lcc gene transcription) and p-coumaric acid selectively induced expression of lcc1 and lcc2; ferulic acid induced lcc3 expression, while 3,5 dihydroxybenzoic acid had no marked effect on laccase gene transcription. The results demonstrate that close-related aromatic compounds appear to have different effects on both laccase activity levels and lcc gene expression in this basidiomycete. PMID- 15341918 TI - Simultaneous silencing of multiple genes in the apple scab fungus, Venturia inaequalis, by expression of RNA with chimeric inverted repeats. AB - RNA-mediated gene silencing has been demonstrated in plants, animals, and more recently in filamentous fungi. Here, we report high frequency, RNA-mediated gene silencing in the apple scab fungus, Venturia inaequalis. The green fluorescent protein (GFP) transgene was silenced in a GFP-expressing transformant. An endogenous gene, trihydroxynaphthalene reductase (THN), involved in melanin biosynthesis, was also silenced. Silencing of these two genes resulted in obvious phenotypes in vitro. High frequency gene silencing was achieved using hairpin constructs for the GFP or the THN genes transferred by Agrobacterium (71 and 61%, respectively). THN-silenced transformants exhibited a distinctive light brown phenotype and maintained the ability to infect apple. Of significance was the simultaneous silencing of the two genes from a single chimeric, inverted repeat hairpin construct. Silencing of both genes with this construct occurred at a frequency of 51% of all the transformants. All 125 colonies silenced for the GFP gene were also silenced for THN. As THN and GFP silenced transformants have readily detectable phenotypes, the genes have utility as markers for gene silencing. Simultaneous, multiple gene silencing, utilising such marker genes, will enable the development of high through-put screening for functional genomics. This chimeric technology will be particularly valuable when linked with silenced genes that have no obvious phenotype in vitro. PMID- 15341919 TI - Loss of SHP-1 phosphatase alters cytokine expression in the mouse hindbrain following cochlear ablation. AB - Inflammatory cytokines in the central nervous system are largely modulated by glial cells and influence neuronal responses to CNS injury. The protein tyrosine phosphatase SHP-1, an intracellular regulator of many cytokine signaling pathways, has been implicated in mediating the activation of glia. There is a direct correlation between abnormally activated microglia and neuron loss within the SHP-1 deficient motheaten (me/me) mouse auditory brainstem after afferent injury. In order to determine whether loss of SHP-1 creates an aberrant cytokine environment driving the abnormal activation of me/me microglia, the expression of interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) was examined by enzyme-linked immunosorbent assay (ELISA). Normal uninjured me/me mice showed lower IL-10 but higher IL-1beta levels compared to wild-type. Following unilateral cochlear ablation, there is decreased expression of IL-4 and IL-10 in me/me brains compared to wild-type, but IL-1beta is significantly increased. These findings indicate that decreases in anti-inflammatory cytokines, in combination with increased expression of the pro-inflammatory cytokine IL-1beta, may initiate a robust inflammatory reaction within the me/me brain contributing to the neuronal degeneration in the deafferented me/me auditory brainstem. SHP-1 may therefore play a role in limiting CNS inflammation following injury and disease. PMID- 15341920 TI - Effect of antioxidants on apoptosis and cytokine release in fetal rat Type II pneumocytes exposed to hyperoxia and nitric oxide. AB - The response of the fetal rat Type II pneumocyte (FTIIP), the stem cell of the alveolar epithelium, to hyperoxia would be helpful to understand the effects of oxygen-induced injury to the developing lung. Our goals were to evaluate the effect of antioxidants (AO) on apoptosis and release of cytokines in freshly isolated FTIIP (day-19) in the presence of 95% O2 and/or nitric oxide (NO). There was increased apoptosis in FTIIP exposed to hyperoxia alone and in combination with NO; this was significantly attenuated (p < 0.01) in the presence of 3 AO, namely grape seed proanthocyanidin extract (GSPE), superoxide dismutase (SOD) and catalase. The anti-inflammatory cytokine IL-10 has been shown to have a role in ameliorating tissue damage owing to persistent inflammation. The release of IL-10 was significantly decreased (p < 0.01) in the presence of GSPE and catalase, compared to control. Addition of SOD led to increased IL-10 compared to GSPE or catalase (p < 0.01) or the combination of GSPE + SOD + catalase (p < or = 0.01). Thus, in our in vitro model of hyperoxic and NO mediated injury to FTIIP, protection from apoptotic cell death with the addition of AO was associated with varying levels of IL-10 release. Our data suggest that the use of SOD and/or IL 10 may decrease hyperoxic lung injury by decreasing apoptosis. Further studies are needed to understand the mode of protection from catalase and GSPE. PMID- 15341921 TI - IL-1beta induction of IL-6 and LIF in normal articular human chondrocytes involves the ERK, p38 and NFkappaB signaling pathways. AB - Interleukin-1 (IL-1) is an important catabolic cytokine in rheumatoid and osteoarthritic joint disease. Besides inducing a catabolic response in articular chondrocytes it also strongly induces synergistic mediators such as leukemia inhibitory factor (LIF) and interleukin-6 (IL-6). The molecular basis of this is so far hardly understood. The aim of our study was to evaluate in vitro and in vivo whether IL-6 and LIF are differentially expressed in normal human and osteoarthritic adult articular chondrocytes and to investigate the potential intracellular signaling pathways of IL-1 involved in these gene regulation events. IL-6 and LIF mRNA expressions were found only at low levels in normal adult articular cartilage. Neither IL-6 nor LIF was strongly over-expressed in osteoarthritic cartilage degeneration. Clearly, both IL-6 and LIF can be very efficiently induced by IL-1beta in articular chondrocytes in vitro. However, this induction was somewhat less in osteoarthritic cells, which were overall activated in terms of expression of both cytokines without stimulation. Experiments using pathway selective inhibitors showed that intracellular signaling of IL-1beta for IL-6 and LIF is mediated by a mixture of the IL-1 signaling cascades. However, the ERK-pathway appeared to be particularly important and might be, therefore, of particular potential if one intends to block induction of these molecules by IL-1 in arthritic joint disease. PMID- 15341922 TI - Cytokine mRNA expression in B cells from bovine leukemia virus-infected cattle with persistent lymphocytosis. AB - We have characterized the expression of six cytokine mRNAs in highly purified B cells from bovine leukemia virus (BLV)-infected cows with persistent lymphocytosis. Selected cytokine mRNAs included those encoding tumor necrosis factor (TNF), lymphotoxin-alpha (LT-alpha), transforming growth factor-beta1 (TGF beta1), interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and interleukin-10 (IL 10). Fresh B cells from cows with persistent lymphocytosis constitutively transcribed TNF, LT-alpha and TGF-beta1 mRNAs. Although IL-1beta, IL-6 and IL-10 mRNAs were barely detectable in fresh B cells from cows with persistent lymphocytosis, transcripts encoding these cytokines were strongly and rapidly upregulated in B cells after cell culture. Results from this study provide the first evidence that B cells infected with BLV express specific cytokine mRNAs in vivo. PMID- 15341923 TI - Gene polymorphisms in pro-inflammatory cytokines are associated with systemic inflammation in patients with severe periodontal infections. AB - The inflammatory response to chronic infections such as periodontitis may be central to the systemic implications of these diseases. This study examined the possible association between specific gene polymorphisms and the systemic inflammatory response in individuals suffering from severe generalized periodontitis. Ninety-four subjects with periodontitis were genotyped for polymorphisms in IL-1A (-889), IL-1B (-511, +3954), TNF-A (-308), IL-6 (-174) and TLR4 (-299, -399) genes. We found that the genotypes for IL-1A or IL-6 are associated with higher levels of serum IL-6 (P < 0.03) and serum CRP (P < 0.05), similarly the TNF-A genotype is associated with higher levels of serum IL-6 (P < 0.05) after correction for age, body mass index, gender, ethnicity and cigarette smoking. Systemic inflammatory responses are higher in severe periodontitis patients carrying rare alleles for functional inflammatory gene polymorphisms. These results suggest that cytokine genotypes are important determinants of the systemic inflammatory response in subjects with periodontitis. Genetic polymorphism therefore, may in part explain the reported association between periodontitis and systemic disease. PMID- 15341924 TI - Advanced glycation end products upregulate angiogenic and pro-inflammatory cytokine production in human monocyte/macrophages. AB - Glucose can react non-enzymatically with amino groups of, for example, proteins, to yield derivatives termed advanced glycation end products (AGE), which contribute to many chronic progressive diseases associated with microvascular complications. The study aimed to determine the effect of AGE-modified albumin on THP-1 cells and human monocyte-derived macrophages. Bovine serum albumin (BSA) or human serum albumin (HSA), modified by glucose-derived AGE, was prepared by incubation with glucose for differing periods of time. Alternatively, BSA was incubated with sodium cyanoborohydride and glyoxylic acid to produce N(epsilon) (carboxymethyl)lysine-modified BSA (CML-BSA). Stimulation for 24h of THP-1 cells with BSA, incubated for 6-8 weeks with glucose, induced significant VEGF release. Human monocyte-derived macrophages stimulated with extensively glycated HSA also showed significant VEGF release, as well as upregulation of IL-8 production, incubation for 6h with extensively glycated HSA increased release of TNFalpha and expression of tissue factor. Finally, addition of CML-BSA resulted in significant induction of TNFalpha and VEGF release. We demonstrate that a range of different methods of glycation of BSA and HSA, including CML-BSA, resulted in the induction of VEGF, TNFalpha, IL-8 and expression of tissue factor, according to length of stimulation and different glycation products used, suggesting that AGE-induced activation of macrophages may contribute to vascular complications by regulation of angiogenic, inflammatory and pro-coagulant processes. PMID- 15341925 TI - Photoactivation of pheophorbide a induces a mitochondrial-mediated apoptosis in Jurkat leukaemia cells. AB - The mechanism of cell death by pheophorbide a (Pba) which has been established to be a potential photosensitizer was examined in experimental photodynamic therapy (PDT) on Jurkat cells, a human lymphoid tumor cell line. In 30-60 min after irradiation, Pba treated cells exhibited apoptotic features including membrane blebbing and DNA fragmentation. Pba/PDT caused a rapid release of cytochrome c from mitochondria into the cytosol. Sequentially, activation of caspase-3 and the cleavage of poly ADP-ribose polymerase (PARP) were followed. Meanwhile, no evidence of activation of caspase-8 was indicated in the cells. In experiments with caspase inhibitors, it was found that caspase-3 alone was sufficient initiator for the Pba-induced apoptosis of the cells. Pba specific emission spectra were confirmed in the mitochondrial fraction and the light irradiation caused a rapid change in its membrane potential. Thus, mitochondria were entailed as the crucial targets for Pba as well as a responsible component for the cytochrome c release to initiate apoptotic pathways. Taken together, it was concluded that the mode of Jurkat cell death by Pba/PDT is an apoptosis, which is initiated by mitochondrial cytochrome c release and caspase-3-pathways. PMID- 15341926 TI - Two modes of the light-induced phytochrome A decline--with and without changes in the proportion of its isoforms (phyA' and phyA''): evidence from fluorescence investigations of mutant phyA-3D pea. AB - Different modes of the phytochrome function are connected with its polymorphism, the major isoforms being phytochromes A and B (phyA and phyB). In its turn, phyA comprises two native species, phyA' and phyA'', whose precise nature and functions remain obscure. With the use of in situ fluorescence spectroscopy, we investigated their properties in a mutant of pea, phyA-3D, characterized by exaggerated photoresponses and impaired photodestruction of phyA. The mutation is a substitution of alanine by valine at the position 194 in phyA. The phyA-3DphyB and phyB mutants were also investigated. In dark-grown plants, all the lines had the content and properties of the two phyA species very similar to the wild type. However, a considerably more intense reduction in [phyA] without changes in the phyA'/phyA'' equilibrium was found in far-red grown mutant plants suggesting a hypersensitivity of phyA-3D with regard to its autoregulation. On the contrary, under red illumination, a higher stability of phyA-3D was observed confirming our earlier findings. This allows a conclusion that the A194V substitution in phyA-3D not only impairs its destruction but also enhances its signaling ability, suggesting a role of this locus in modulation of its activity. PMID- 15341927 TI - Intranasal irradiation with the xenon chloride ultraviolet B laser improves allergic rhinitis. AB - We earlier reported that the 308 nm xenon chloride (XeCl) ultraviolet B (UVB) laser is highly effective for the treatment of inflammatory skin diseases. Since UVB irradiation has been shown to exert both local and systemic immunosuppression, we investigated the clinical efficacy of UVB irradiation in allergic rhinitis. In an open study, groups of patients with severe allergic rhinitis received intranasal irradiation with a 308 nm XeCl UVB excimer laser for two weeks. In the low-dose group (n=10), treatment was given twice weekly, starting with 0.25x the individual minimal erythema dose (MED), whereas patients in the medium-dose group (n=8) were treated four times weekly, starting with 0.4x MED. In each group, the dosage was gradually increased. Evaluation was based on the symptom scores. The effect of the XeCl laser on the skin prick test reaction was also studied. In the low-dose group, seven patients completed the study, and there was no improvement in the nasal symptoms. In the medium-dose group, the XeCl UVB irradiation significantly inhibited the rhinorrhoea, the sneezing, the nasal obstruction and the total nasal score (p<0.05). The XeCl UVB excimer laser also inhibited the allergen-induced skin prick test in a dose-dependent manner. These results suggest that the XeCl UVB excimer laser might serve as a new therapeutic tool in the treatment of allergic rhinitis. PMID- 15341928 TI - Daily variation in cellular content of UV-absorbing compounds mycosporine-like amino acids in the marine dinoflagellate Scrippsiella sweeneyae. AB - Sudden exposure experiments to high PAR (photosynthetically available radiation) or high PAR+UVR (ultraviolet radiation) were conducted for the marine dinoflagellate Scrippsiella sweeneyae acclimated to either low PAR or high PAR to determine the induction of cellular mycosporine-like amino acid (MAA) in relation to photosynthesis status. When the exposure to high PAR (30.8 Wm(-2)) was provided at different time in the light period for S. sweeneyae acclimated to low PAR (7.7 Wm(-2)) which suppressed photosynthesis, S. sweeneyae could enhance the induction of MAA but it only occurred in the first half of the light period. When UVR exposure was provided for the culture acclimated to high PAR which enhanced photosynthesis, cellular MAA content did not increase during the entire light period, but displayed daily variation similar to the control for two and half days. Daily variation of cellular MAA content did not synchronized with that of cell volume and cellular chlorophyll a content. The individual MAAs also revealed similar daily variations with different phase, which increased for a few hours in the beginning of the light period, except for cellular palythine content. Thus the total cellular MAA content revealed daily variation with changing the relative composition within a few hours. As one of the biological protective strategies against harmful UVR in sunlight, the daily vertical migration in the bloom forming dinoflagellates might be accompanied by the daily variation of cellular MAA content for a photosynthesis at daytime. PMID- 15341929 TI - A method for separating ALA from ALA derivatives using ionic exchange extraction. AB - Photodynamic therapy using 5-aminolevulinic acid (ALA)-induced protoporphyrin IX is a recent approach to detect and treat some malignancies. The use of lipophilic derivatives of ALA has been exploited in the last years to enhance ALA penetration. In this paper, we describe the application of the Mauzerall and Granick's method [J. Biol. Chem. 219 (1956) 435] to the quantification of ALA derivatives. We also describe the employment of reusable ion-exchange chromatographic columns for separating mixtures of ALA and ALA derivatives present in biological samples. The relation between 555 nm absorbance and ALA or ALA derivative concentration was linear up to 100 nmol/ml and the limit of detection of ALA and ALA derivatives was 1 nmol per ml. We employed a Dowex 50 X8 hydrogen form resin to separate ALA from the derivatives. Whereas 90+/-4% of the total ALA was eluted using sodium acetate, only 3-9% of the ALA derivatives was recovered. Only upon exposure of the resin to a high HCl concentration, the ALA derivatives were completely released. We employed this new method for the separation of ALA from ALA derivatives in cells exposed to different ALA compounds. PMID- 15341930 TI - Inhibition of the strand transfer step of HIV-1 integrase by non-natural dinucleotides. AB - New, non-natural dinucleotide 5'-monophosphates, with a surrogate isonucleoside component of l-related stereochemistry at the 'terminal' position, have been synthesized. Structures of 2a-c were confirmed by multinuclear NMR spectra ((1)H, (13)C, (31)P, COSY), UV hypochromicity and FAB HRMS data. These compounds are totally resistant to cleavage by 3'- and 5'-exonucleases. The dinucleotides showed remarkable selectivity for inhibition of the strand transfer step of HIV-1 integrase. To the best of our knowledge, these compounds represent only the second example of selective strand transfer inhibitors of HIV integrase. PMID- 15341931 TI - Novel pyrazinone inhibitors of mast cell tryptase: synthesis and SAR evaluation. AB - In this manuscript, the synthesis and SAR evaluation of a novel pyrazinone class of tryptase inhibitors is described. Chemical optimization of the P1 and P4 groups led to the identification of 7p (K(i)=93 nM) as a potent inhibitor of mast cell tryptase. PMID- 15341932 TI - Computational comparison of microtubule-stabilising agents laulimalide and peloruside with taxol and colchicine. AB - Microtubule-stabilising agents laulimalide and peloruside have been compared with tubulin-interacting drugs paclitaxel and colchicine by different computational approaches. Docking and QSAR-based programs point to a favourable interaction with the beta tubulin paclitaxel binding site, although an additional, preferred binding site has been found at the alpha subunit of tubulin. All together provides a plausible rationalisation of the singular binding features of these microtubule stabilisers and paves the way for future structural studies. PMID- 15341933 TI - Studies on adenosine A2a receptor antagonists: comparison of three core heterocycles. AB - Piperazine and (R)-2-(aminomethyl)pyrrolidine derivatives of [1,2,4]triazolo[1,5 a][1,3,5]triazine have recently been shown to be potent and selective adenosine A(2a) receptor antagonists. We have replaced the triazolotriazine core structure with two different heterocyclic cores. One of these, the one deriving from [1,2,4]triazolo[1,5-c]pyrimidine, appears to be particularly effective and selected analogs from this series have been shown to be orally active in a mouse catalepsy model of Parkinson's disease. PMID- 15341934 TI - Triamino derivatives of triazolotriazine and triazolopyrimidine as adenosine A2a receptor antagonists. AB - Piperazine derivatives of 2-furanyl[1,2,4]triazolo[1,5-a][1,3,5]triazine have recently been shown to be potent and selective adenosine A(2a) receptor antagonists. We now demonstrate that potent and selective A(2a) receptor antagonists could still be obtained when the arylpiperazines are separated from the triazolotriazine core structure by an ethylenediamine spacer. Selected analogs bearing this triazolotriazine or the related triazolopyrimidine core structure have been found to be orally active in a mouse catalepsy model of Parkinson's disease. PMID- 15341935 TI - End-capping of the modified melanocortin tetrapeptide (p-Cl)Phe-D-Phe-Arg-Trp-NH2 as a route to hMC4R agonists. AB - Of the 42 R'-X-(p-Cl)Phe-D-Phe-Arg-Trp-NH(2) (X=CO, SO(2), PO, PS) tested at the human (h)MC1, hMC3, and hMC4 receptors (R), the most potent MC4R agonists (EC(50) of 8-20 nM) were obtained by end-capping with R'=CH(2)CHCH(2) (9), NCCH(2) (16), NH(2)COCH(2) (17), HCONHCH(2) (18), CH(3)NH (19), CH(2)CHCH(2)NH (21), 2-Th (23), PhCH(2) (30) and X=CO. These compounds possess 35-60-fold hMC4 versus hMC1Rs selectivity with urea LK-71 (19) being the most potent at hMC4R and MC4/1R selective (EC(50)=8.5 nM, MC4/1R=100). LK-75 (16) combines high potency at hMC4R and MC4/3R selectivity (EC(50)=10.5 nM, MC4/3R=290). SAR is discussed. PMID- 15341936 TI - Modeling the binding affinities of beta-secretase inhibitors: application to subsite specificity. AB - A new linear binding affinity model has been developed for hydroxyethylene based inhibitors of beta-secretase (BACE). This model is an improvement over a previously published model, and has been applied to a series of analogs not included in the training set. The linear model has been used to study subsite specificity for the P(2) through P(2)' positions, and to evaluate a small number of C-terminal analogs. The predicted rankings are in good agreement with experiment and support using this model for structure-based design of BACE inhibitors. PMID- 15341937 TI - N-(1-Benzylpyrrolidin-3-yl)arylbenzamides as potent and selective human dopamine D4 antagonists. AB - A series of N-(1-benzylpyrrolidin-3-yl)arylbenzamides 8 has been prepared, and their structure-activity relationships studied. Potent ligands selective for human D(4) (hD(4)) over hD(2) and alpha(1) have been identified. One example was determined to be an antagonist in a cAMP assay, with an IC(50) of 1500 nM. PMID- 15341938 TI - Design and synthesis of 3'-ureidoadenosine-5'-uronamides: effects of the 3' ureido group on binding to the A3 adenosine receptor. AB - On the basis of high binding affinity at the A(3) adenosine receptor of 3' aminoadenosine derivatives with hydrogen bonding donor ability, novel 3' ureidoadenosine analogues were synthesized from 1,2:5,6-di-O-isopropylidene-d glucose in order to lead to stronger hydrogen bonding than the corresponding 3' aminoadenosine derivatives. However, the synthesized 3'-ureidoadenosine analogues were totally devoid of binding affinity, because 3'-urea moiety caused steric and electrostatic repulsions at the binding site of the A(3) adenosine receptor, leading to conformational distortion. PMID- 15341939 TI - New DNA binding ligands as a model of chromomycin A3. AB - Small molecules with DNA-binding affinity within the minor groove have become of great interest. In this study, new DNA-binding ligands were designed to mimic Chromomycin A(3) (CRA(3)), which contains a hydroxylated tetrahydroanthracene chromophore substituted with di and trisaccharides. The trisaccharide part of CRA(3) that is supposed to contribute to form the Mg(2+)-coordinated dimer was expected to be mimicked by a simple alkyl group attached to the chromophore part as new model compounds. The present study has successfully demonstrated that the new ligands form Mg(2+)-coordinated dimer complexes to exhibit DNA-binding affinity. PMID- 15341940 TI - Design and synthesis of conformationally constrained 3-(N alkylamino)propylphosphonic acids as potent agonists of sphingosine-1-phosphate (S1P) receptors. AB - A series of conformationally constrained 3-(N-alkylamino)propylphosphonic acids were systematically synthesized and their activities as S1P receptor agonists were evaluated. Several pyrrolidine and cyclohexane analogs had S1P receptor profiles comparable to the acyclic lead compound, 3-(N tetradecylamino)propylphosphonic acid (3), lowered circulating lymphocytes in mice after iv administration and were thus identified as being suitable for further investigations. PMID- 15341941 TI - Identification of 4-piperazin-1-yl-quinazoline template based aryl and benzyl thioureas as potent, selective, and orally bioavailable inhibitors of platelet derived growth factor (PDGF) receptor. AB - 4-[4-(N-Substituted-thio-carbamoyl)-1-piperazinyl]-6-methoxy-7-alkoxyamino quinazoline derivatives such as 14 (CT53986) have been identified to be potent and selective inhibitors of the phosphorylation of PDGFR. SAR-investigations are described in the arylamine segment, C-7 appendage, and the thiourea moiety. Bioisosteres of thiourea (cyanoguanidine), and of quinazoline (quinoline-3 carbonitrile) were synthesized and are compared for their in vitro inhibitory activity. PK profiles of the optimized compounds in rat, dog, and cynomolgus monkey are described. PMID- 15341942 TI - Synthesis and evaluation of 2-amino-8-alkoxy quinolines as MCHr1 antagonists. Part 1. AB - A high-throughput screen was performed in order to identify chemotypes that are bound by the melanin concentrating hormone receptor-1 (MCHr1). A novel 2-amino-8 alkoxyquinoline compound (1) was identified and subsequently optimized using a parallel and automated procedure for the rapid production of multiple analogs. The structure-activity relationships that emerged from this effort are described, along with selected pharmacokinetic parameters of compound (d)-61 when dosed orally in diet-induced obese mice. PMID- 15341943 TI - Synthesis and evaluation of 2-amino-8-alkoxy quinolines as MCHr1 antagonists. Part 2. AB - The continued SAR investigation of 2-amino-8-alkoxy quinolines as melanin concentrating hormone receptor-1 (MCHr1) antagonists is reported. Prior hit-to lead efforts resulted in the identification of 1 as a robust MCHr1 antagonist. Further delineation of the structural parameters essential for MCHr1-binding affinity of this class of nontraditional GPCR ligands resulted in the identification of compounds such as 33, 34 and 37, which demonstrate single digit nanomolar antagonism of MCHr1-mediated Ca(2+) release. The synthesis and biological evaluation of these compounds are reported. PMID- 15341944 TI - Synthesis and evaluation of 2-amino-8-alkoxy quinolines as MCHr1 antagonists. Part 3. AB - Prior SAR studies on 2-amino-8-alkoxyquinoline MCHr1 antagonists demonstrated that compounds with acyclic amide-containing sidechains displayed exceptional binding and functional potency, but negligible CNS penetration. Related analogs with acyclic benzylamine-containing sidechains showed greatly improved CNS exposure, but suffered in functional potency. In this report, we demonstrate that cyclization of these benzylic amine sidechains affords compounds that combine the best elements of potency and CNS penetration among this class of antagonists. This is exemplified by compound 21, which has sub-nanomolar MCHr1 binding affinity, good functional potency, and excellent CNS exposure over 24h. PMID- 15341945 TI - Unnatural base pairs between 2- and 6-substituted purines and 2-oxo(1H)pyridine for expansion of the genetic alphabet. AB - An unnatural base pair between 2-amino-6-(2-thienyl)purine (denoted by s) and 2 oxo(1H)pyridine (denoted by y) shows high selectivity in transcription and translation. Toward the further development of unnatural base pairs that also have exclusive selectivity in replication, we examined the roles of the 2-amino and 6-thienyl groups of s using base pairs between y and purine-analogs, 6 thienylpurine and 2-amino-6-furanylpurine, as well as s. The results obtained from the thermal stability and DNA polymerase single-nucleotide insertion experiments suggest that the 2-amino group of s contributes toward the shape complementarity of the pairing with y, rather than the hydrogen bonding with the 2-keto group of y. In addition, the bulkiness of positions 2 and 6 of the unnatural purines cooperatively determines the selectivity of the noncanonical pairing with y or the natural pyrimidines in replication. This information is useful not only for the development of unnatural, orthogonal base pairs, but also for understanding the mechanisms of base pair formation in replication. PMID- 15341946 TI - Discovery of orally active prostaglandin D2 receptor antagonists. AB - A series of N-(p-alkoxy)benzoyl-2-methylindole-4-acetic acids were synthesized and evaluated for prostaglandin D(2) (DP) receptor affinity and antagonist activity. Some of them exhibited strong receptor binding and were potent in the cAMP formation assays. These antagonists also suppressed allergic inflammatory responses such as the PGD(2)-induced increase of microvascular permeability. Structure-activity relationship (SAR) data are presented. PMID- 15341947 TI - Potent and selective P2-P3 ketoamide inhibitors of cathepsin K with good pharmacokinetic properties via favorable P1', P1, and/or P3 substitutions. AB - A series of ketoamides were synthesized and evaluated for inhibitory activity against cathepsin K. Exploration of the interactions between achiral P(2) substituents and the cysteine protease based on molecular modelling suggestions resulted in potent cathepsin K inhibitors that demonstrated high selectivity versus cathepsins B, H, and L. Subsequent modifications of the P(3), P(1), and P(1') moieties afforded orally bioavailable inhibitors. PMID- 15341948 TI - Synthesis of benzimidazole based analogues of sphingosine-1-phosphate: discovery of potent, subtype-selective S1P4 receptor agonists. AB - Sphingosine-1-phosphate (S1P) is a biologically active lysophospholipid with the capacity to induce a broad range of cellular responses via its interaction with the S1P family of G-protein coupled receptors. A member of this receptor family, S1P(4), is highly and almost exclusively expressed in the lymphoid system and has been implicated in regulation of cell shape and motility. This report describes the synthesis of several potent benzimidazole based S1P(4) receptor selective agonists. For instance, compound 9b displayed an EC(50)=36 nM at the S1P(4) receptor using a [gamma-(35)S]GTP binding assay as compared to an EC(50)=37 nM for the endogenous ligand. We also report the effects of altering stereochemistry at the C2 position, methylation at the C1 and C2 position, and activity differences between the alcohol and phosphate head groups of the analogues. PMID- 15341949 TI - Pyrrolo[2,1-c][1,4]benzodiazepine-anthraquinone conjugates. Synthesis, DNA binding and cytotoxicity. AB - New pyrrolobenzodiazepine-anthraquinone hybrids have been designed and synthesized, found to effectively bind to DNA and also exhibit cytotoxicity against many cancer cell lines PMID- 15341950 TI - A new class of acyclic 2-alkyl-1,2-diaryl (E)-olefins as selective cyclooxygenase 2 (COX-2) inhibitors. AB - A new class of (E)-2-alkyl-2-(4-methanesulfonylphenyl)-1-phenylethenes were designed for evaluation as selective cyclooxygense-2 (COX-2) inhibitors. The target olefins were synthesized, via a Takeda olefination reaction, followed by oxidation of the respective thiomethyl olefinic intermediate. In vitro COX-1/COX 2 inhibition studies identified (E)-2-(4-methanesulfonylphenyl)-1-phenyloct-1-ene (8d) as a potent (IC(50)=0.77 microM) and selective (Selectivity Index>130) COX-2 inhibitor. PMID- 15341951 TI - The identification and optimization of 2,4-diketobutyric acids as flap endonuclease 1 inhibitors. AB - There have been several recent reports of chemopotentiation via inhibition of DNA repair processes. Flap endonuclease 1 (FEN1) is a key enzyme involved in base excision repair (BER), a primary pathway utilized by mammalian cells to repair DNA damage. In this report, we describe the identification and SAR of a series of 2,4-diketobutyric acid FEN1 inhibitors. PMID- 15341952 TI - Synthesis and biological evaluation of novel cytotoxic phospholipids for prostate cancer. AB - We describe herein synthesis, SAR, and biological evaluation of a novel series of cytotoxic serine amide phosphates (SAPs) for prostate cancer. These compounds were tested for their cytotoxicity in five human prostate cancer cell lines (DU 145, PC-3, LNCaP, PPC-1, and TSU), and in CHO and RH7777 cells (negative controls). Comparison of anticancer effects of these compounds with a standard chemotherapeutic agent 5-fluorouracil shows that they are very effective in killing prostate cancer cells with low micromolar cytotoxicity and provide us a new lead for the development of drugs for prostate cancer. PMID- 15341953 TI - 7-Substituted 2-phenyl-benzofurans as ER beta selective ligands. AB - A series of 2-(4-hydroxy-phenyl)-benzofuran-5-ols with relatively lipophilic groups in the 7-position of the benzofuran was prepared and the affinity and selectivity for ER beta was measured. Many of the analogues were found to be potent and selective ER beta ligands. Additional modifications at the benzofuran 4-position as well as at the 3'-position of the 2-phenyl group were found to further increase selectivity. Such modifications led to compounds with <10 nM potency and >100-fold selectivity for ER beta. PMID- 15341954 TI - Synthesis and evaluation of 4-(1-aminoalkyl)-N-(4-pyridyl)cyclohexanecarboxamides as Rho kinase inhibitors and neurite outgrowth promoters. AB - The influence of stereochemistry and alkyl side chain length on the bioactivity of the Rho kinase inhibitor Y-27632 [(+)-1, R=Me] was examined by the synthesis of (+)- and (-)-1, and two alkyl chain analogs (+)- and (-)-2 (R=n-propyl) and ( )-3 (R=n-octyl) as well as their evaluation in enzymatic and neurite outgrowth assays. PMID- 15341955 TI - Synthesis and evaluation of novel oxazoline MMP inhibitors. AB - MMP inhibitors with novel oxazoline zinc binding groups have been synthesized and evaluated. Selectivity for the inhibition of MMP-9 over MMP-1, MMP-2, and MMP-12 has been achieved in several cases. PMID- 15341956 TI - Effective inhibition of HIV-1 replication in cultured cells by external guide sequences and ribonuclease P. AB - We examined the suppressive effect of HIV-1 RNA gene cleavage on HIV-1 expression, using the catalytic RNA subunit RNase P and the 3'-half tRNA(Try) [external guide sequence (EGS)] in cultured cells. HIV-1 expression was inhibited by the tRNA(met)-EGS-U5 and U6-EGS-U5 from the tRNA(met) and U6 promoters, respectively. There was no difference in the inhibitory effects on HIV-1 expression between the tRNA(met) and U6 promoters. PMID- 15341957 TI - The development of new bicyclic pyrazole-based cytokine synthesis inhibitors. AB - 4-Aryl-5-pyrimidyl-based cytokine synthesis inhibitors of TNF-alpha production, which contain a novel bicyclic pyrazole heterocyclic core, are described. Many of these inhibitors showed low nanomolar activity against LPS-induced TNF-alpha production in a THP-1 cell-based assay and against human p38 alpha MAP kinase in an isolated enzyme assay. The X-ray crystal structure of a bicyclic pyrazole inhibitor co-crystallized with mutated p38 (mp38) is presented. PMID- 15341958 TI - Synthesis and GABA receptor potency of 3-thiomethyl-4-(hetero)aryl-5-amino-1 phenylpyrazoles. AB - A convenient synthetic route to novel 4-arylpyrazoles is described. The potential for insecticidal activity through GABA channel blockage by this series of compounds, as well as their selectivity for insect versus mammalian receptors, are explored through in vitro and in vivo assays. PMID- 15341959 TI - Study on noncovalent complexes of cytotoxic protoberberine alkaloids with double stranded DNA by using electrospray ionization mass spectrometry. AB - The noncovalent complexes of four cytotoxic protoberberine alkaloids that is, berberine, palmatine, jatrorrhizine, and coptisine with double-stranded oligodeoxynucleotides d(AAGAATTCTT)(2) were investigated by electrospray ionization mass spectrometry. These four active components from Chinese herbal medicines showed both 1:1 and 1:2 binding stoichiometries, independent on the alkaloid-to-DNA ratios. Binding affinities in the order of palmatine> or =jatrorrhizine>coptisine>berberine with d(AAGAATTCTT)(2) were obtained. Additionally, the preliminary results indicated that berberine had some sequence selectivities. PMID- 15341960 TI - Synthesis, structural properties and insulin-enhancing potential of bis(quercetinato)oxovanadium(IV) conjugate. AB - An oxovanadium complex of quercetin (2), exhibits highly potent insulin-enhancing activity in streptozotocin-induced diabetic mice. It also mimics mitogenic potential of insulin as evaluated by [H(3)]thymidine uptake assay making an effective, orally active insulin-enhancing agent for the treatment of both type 1 and type 2 diabetes without any noticeable toxic effects. PMID- 15341961 TI - Synthesis and structure-activity relationships of uracil derived human GnRH receptor antagonists: (R)-3-[2-(2-amino)phenethyl]-1-(2,6-difluorobenzyl)-6 methyluracils containing a substituted thiophene or thiazole at C-5. AB - The synthesis of a series of (R)-3-[2-(2-amino)phenethyl]-1-(2,6-difluorobenzyl) 6-methyluracils containing a substituted thiophene or thiazole at C-5 is described. SAR around C-5 of the uracil led to the discovery that a 2-thienyl or (2-phenyl)thiazol-4-yl group is required for optimal receptor binding. The best compound from the series had a binding affinity of 2 nM (K(i)) for the human GnRH receptor. A novel and convenient preparation of N-1-(2,6-difluorobenzyl)-6 methyluracil is also described. PMID- 15341962 TI - Steroid and lipid conjugates of siRNAs to enhance cellular uptake and gene silencing in liver cells. AB - Double-stranded short interfering RNAs (siRNAs) mediate post-transcriptional inhibition of gene expression in a variety of biological systems. However, human liver cells show poor uptake of these nucleic acids. In order to improve the delivery of siRNA into these cells without transfection agents, we have synthesized two series of lipophilic siRNAs conjugated with derivatives of cholesterol, lithocholic acid or lauric acid. The lipid moieties were covalently linked to the 5'-ends of the RNAs using phosphoramidite chemistry. The potency of these chemically modified siRNAs to inhibit reporter gene expression was further investigated in vitro with beta-galactosidase expressing liver cells. PMID- 15341963 TI - Synthesis of acyloxymethyl ester prodrugs of the transferable protein farnesyl transferase substrate farnesyl methylenediphosphonate. AB - Three isoprenoid diphosphate analogues of farnesyl diphosphate (FPP) where the diphosphate has been replaced by methylene diphosphonate and the negative charges masked by frangible pivaloyloxymethyl (POM) esters were prepared. Farnesyl methylenediphosphonate is a sub-micromolar substrate for protein farnesyl transferase. The tripivaloyloxymethyl esters of isoprenoid methylenediphosphonate have significantly increased lipophilicity and may act as important farnesyl diphosphate prodrugs. PMID- 15341964 TI - Synthesis of dimeric acridine derived antivirals. AB - A series of antiviral compounds consisting of an intercalating acridine derived part, a spacer region and a reactive EDTA derived conjugate was synthesized in an easy sequence starting from 1,omega-alkyldiamines. As shown in model screenings, in the presence of ascorbic acid the Fe-complexes of these compounds reduced the phage-titer of MS2-phages by >8 logarithmic decades. PMID- 15341965 TI - Asymmetric synthesis of the stereoisomers of 11,12,15(S)-trihydroxyeicosa 5(Z),8(Z),13(E)-trienoic acid, a potent endothelium-derived vasodilator. AB - The four stereoisomers of the endothelial-derived vasorelaxant 11,12,15(S) trihydroxyeicosatrienoic acid [1, 11,12,15(S)-THETA] were prepared by a triply convergent, asymmetric route that exploited the stereospecific, copper mediated cross-coupling of alpha,beta-dialkoxystannanes with organic electrophiles and the utility of dialkylthionocarbamates as orthogonal alcohol protective groups. Only 11(R),12(S),15(S)-THETA was comparable to natural material by HPLC, GC/MS, and in vitro bioassay. PMID- 15341966 TI - Synthesis of enantiomerically pure D-FDOC, an anti-HIV agent. AB - The beta-D-enantiomer of FDOC (2',3'-dideoxy-5-fluoro-oxacytidine) exhibits potent anti-HIV-1 activity. It was obtained in optically pure form by employing a tandem kinetic resolution/chiral salt crystallization protocol. In addition, conditions were developed that allowed the unwanted butyrate ester of the L enantiomer of FDOC to be racemized. This material could then be recycled in future resolutions. PMID- 15341967 TI - Synthesis and vasorelaxant properties of hybrid molecules out of NO-donors and the beta-receptor blocking drug propranolol. AB - S-Nitroso-N-acetylpenicillamine (SNAP) and 3-nitrooxypivaloyl acid were combined in the form of the respective amides with propranolol, in order to obtain prodrugs (NO-propranololes) with beta-receptor blocking properties of the latter compound with nitric oxide releasing properties of the former compounds. A respective nitratoester could not be synthesized, because it immediately rearranges to the amide after deprotection of the amino group. In vitro tests on porcine pulmonary arteries showed that both types of hybrid molecules (6, 12) elicited vasorelaxation, but the nitratoamide was less potent by more than one order of magnitude. The vasorelaxant effect of SNAP was more pronounced than that of the SNAP-hybrid (12), on the other hand the nitratoamide 6 was more potent than 3-nitrooxypivaloyl acid. PMID- 15341968 TI - Estrone formate: a novel type of irreversible inhibitor of human steroid sulfatase. AB - A series of estrone conjugates of the type estrone-3-O-C(O,S)-X have been prepared and evaluated for inhibition of human steroid sulfatase (STS). Among the carbamate (6), thiocarbamate (8), cyanate (7), formate (9), and acetate (10) analogs of estrone, only 9 was found to inhibit STS in a time- and concentration dependent manner. With an IC(50) of 0.42 microM 9 is the first potent inactivator of STS which does not feature the sulfamate group. Furthermore a formate-type inhibitor featuring a benzoxazole moiety in place of the steroid skeleton (14) was prepared, suggesting a general principle of inactivation by the formate group. As the mode of action we propose an immediate transfer of the formyl moiety to a nucleophilic residue in the active site of STS. PMID- 15341969 TI - Synthesis of ether analogues derived from HUN-7293 and evaluation as inhibitors of VCAM-1 expression. AB - The cyclic depsipeptide HUN-7293 (1) and its D-lactate analogue 2 are highly potent inhibitors of inducible cell adhesion molecule expression. We report the synthesis of ether analogues varying in stereochemistry and side chain at the former hydroxyl acid position by employing a 'cut and paste chemistry' methodology starting from 1. As an additional fruit of this synthetic effort, a cyclodepsipeptide featuring a tertiary amine instead of a tertiary amide between PrLEU and MALA was obtained. Results on the inhibitory profile of these compounds in assays of VCAM-1 and ICAM-1 protein expression are discussed. PMID- 15341970 TI - P4 and P1' optimization of bicycloproline P2 bearing tetrapeptidyl alpha ketoamides as HCV protease inhibitors. AB - With the aim of improving HCV protease inhibitors reported in our previous manuscripts, we synthesized and evaluated a series of 1a-based tetrapeptidyl alpha-ketoamides with additional P4 modification. The promising analog discovered through this SAR, 5a, was further derivatized at P1' or P1 position. As a result of these efforts, we found that replacement of the P4 valine as seen in 1a with cyclohexylglycine (Chg) resulted in the discovery of 5a, 5c, and 5e endowed with improved cellular activity in comparison to 1a. PMID- 15341971 TI - Pyridazine derivatives and related compounds. Part 13: Synthesis and antimicrobial activity of some pyridazino[3',4':3,4]pyrazolo[5,1-c]-1,2,4 triazines. AB - A general method for the preparation of substituted pyridazinopyrazolotriazines is reported. 3-Aminopyrazolo[3,4-d]pyridazine was diazotized and coupled with active methylene reagents to afford the tricyclic pyridazino[3',4':3,4]pyrazolo[5,1-c]-1,2,4-triazines with substituents such as methyl, phenyl, ethoxycarbonyl, acetyl or benzoyl, depending on the methylene reagent used. In addition several condensation reactions with hydrazines gave the corresponding acid hydrazide and/or hydrazones. Reactions of 3 with aromatic aldehydes afforded hydrazones. The products were screened for their antimicrobial activity against five microorganisms. PMID- 15341972 TI - Targeting the IGF-1 receptor: from rags to riches. PMID- 15341973 TI - Targeted therapy--how successful has it been? PMID- 15341974 TI - Sentinel node biopsy in gastrointestinal-tract cancer. AB - Forty three years after Gould's first description of the sentinel lymph node (SN) technique in malignant tumours of the parotid, sentinel lymph node biopsy (SLNB) has become an invaluable tool for the treatment of solid tumours. In some tumour types, it has been shown to reliably reflect the lymph node (LN) status of the tumour-draining LN basin. In melanoma and breast cancers, it has become a widely accepted element in the routine surgical management of these malignant diseases. In gastrointestinal tumours, the technique is currently under intense investigation. First reports on its application in other solid tumours like non small cell lung cancer, thyroid carcinoma, oropharyngeal carcinoma, vulvar carcinoma, and Merkel Cell carcinoma of the skin were published more recently. In the following review, we will give a synopsis of the fundamentals of the SN concept and will then proceed to an overview of recent advances of SLNB in gastrointestinal cancers. PMID- 15341975 TI - The treatment of cancer in Greek antiquity. AB - Literary sources provide considerable information on the existence of various malignant tumours in the classical period. Based on a close reading of the ancient Greek medical treatises, this paper traces the history of the treatment of cancer by examining the theories of tumour formation, as they were codified by leading physicians of antiquity, together with the therapeutic methods they proposed in their writings. The discussion focuses on a series of medical texts beginning with the Hippocratic corpus (ca. 460-370 B.C.) and the voluminous works of Galen (129-199 A.D.) and extends to medical handbooks (Oreibasios, Aetios of Amida, Paul of Aegina) composed in subsequent centuries up to the end of the ancient world (VII c. A.D.). PMID- 15341976 TI - Complementary and alternative medical therapies for children with cancer. AB - Complementary and alternative medical therapies (CAM) are treatments that generally fall outside of the mainstream of conventional medicine. CAM therapies are used by 31-84% of children with cancer, including many children enrolled on clinical trials. CAM therapies are often used for the treatment of side-effects of cancer or cancer therapy, and only rarely as an alternative to conventional therapy. Regulation of CAM therapies varies worldwide, and many therapies have not been subject to scientifically conducted analyses. Adverse events have been described, especially from the contamination of herbs. Only rare reports of interactions of CAM therapies with conventional anticancer treatments have been reported. Several research studies of CAM in children with cancer are underway. In the interim, non-pharmacological therapies such as mind-body medicine, manipulative and body-based therapies and energy therapies may be used for supportive therapy. Research is needed before biologically based CAM therapies may be recommended in conjunction with conventional therapy. PMID- 15341977 TI - A clinical phase I trial of gemcitabine and treosulfan in uveal melanoma and other solid tumours. AB - This trial was performed to define the maximum tolerated dose (MTD) of treosulfan administered in combination with a fixed dose of gemcitabine in uveal melanoma patients. Preclinical studies suggested synergistic activity against uveal melanoma. Gemcitabine (1 g/m2) and treosulfan (2.5-4 g/m2) were administered on days 1 and 8, and cycles were repeated on day 29 for a maximum of six cycles. For treosulfan, dose escalation cohorts of 2-4 patients were enrolled. An additional 25 patients were entered at treosulfan dose levels II (3 g/m2) and III (3.5 g/m2). Thirty three patients with uveal melanoma and six patients with other histologies were accrued. Side-effects were predominantly haematological. The MTD was 3.5 g/m2 of treosulfan together with 1 g/m2 of gemcitabine. In the uveal melanoma patients, one partial response (PR) and 15 stablisations of disease (SD) were recorded and whether this translates into a survival gain should be explored further. PMID- 15341978 TI - Woman feels breast lump--surgeon cannot: the role of ultrasound in arbitration. AB - The role of ultrasound scanning (USS) in patients complaining of a breast lump where the clinical examination (CE) is normal is not clearly defined. To determine this in greater detail, all patients complaining of a breast lump underwent CE. Where no lump could be found, but was still reported by the patient, an USS was performed. All lesions underwent biopsy and/or aspiration, as well as mammography in suspicious cases or those over 40 years of age. This cohort represented 5% of all referrals in the study period. Four hundred and twenty women were prospectively studied in this way. Median follow up is 3.4 years (range 2.5-4.2 years). Twenty two had solid lumps (of which 3 were cancers) and 48 had cysts. Nineteen patients re-presented with symptoms in the same breast (median time = 12 months (range 4.5-20 months), all of which were imaged on USS: 15 cysts and 4 further cancers (3 in the same quadrant as the original lump, one contralateral) were identified. Women with symptomatic breast lumps and a normal CE can be considered a reliable indication that cancer is very unlikely to be present (negative predictive value = 0.98). Ultrasound may be a suitable complimentary investigation, which will relieve symptoms in those with cysts and can detect small clinically--and sometimes mammographically--occult breast cancers. PMID- 15341979 TI - Psychological distress in women at increased risk for breast cancer: the role of risk perception. AB - The magnetic resonance imaging screening (MRISC) study evaluates a surveillance programme for women with a hereditary risk for breast cancer. The psychological burden of surveillance in these women may depend on inaccurate risk perceptions. To examine differences in risk perceptions between three predefined risk categories and associations with psychological distress. BC-specific distress, general distress, and RP (cognitive and affective) were assessed, two months before a surveillance appointment. Cumulative lifetime risk (CLTR) of developing breast cancer was trichotomised into: (1) CLTR of 60-85% (mutation carriers), (2) CLTR of 30-50%, and (3) CLTR of 15-30%. In a total group of 351 women (mean age 40.5 years, range 21-63 years) the three risk categories significantly differed in their accuracy of assessing cognitive risk perceptions. In category 1, 60% had an accurate risk perceptions, in category 2, 43.7% and in category 3, 33.3%. Overestimators reported significantly more breast cancer-specific distress. After adding affective risk perception to the model, this effect disappeared. Affective risk perceptions showed significant associations with breast cancer-specific and general distress. Affective risk perception is a more important determinant for psychological distress than cognitive risk perception. This knowledge should be used during surveillance appointments in order to improve and individualise support for these women. PMID- 15341980 TI - The distribution of drug-efflux pumps, P-gp, BCRP, MRP1 and MRP2, in the normal blood-testis barrier and in primary testicular tumours. AB - The drug-efflux pumps P-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1) are present in the blood-testis barrier (BTB) and may hamper the delivery of cytotoxic drugs to the testis. The precise localisation of P-gp and MRP1 in testicular tissue and the presence of the efflux pumps MRP2 and breast cancer resistance protein (BCRP) in the BTB are unknown. We therefore studied the localisation of these pumps in the BTB in normal testis (n = 12), in non-seminoma (n = 10) seminoma (n = 10), and testicular lymphoma (n = 9). Slides were scored semi-quantitatively for P-gp, MRP1, MRP2 and BCRP and blood vessels with factor VIII antibody. In normal testis, P-gp and BCRP were strongly expressed by myoid cells and luminal capillary endothelial wall and P-gp also by Leydig cells. MRP1 was observed at the basal side of Sertoli cells and on Leydig cells. MRP2 was only weakly expressed by myoid cells. Seminomas and non-seminomas expressed P-gp and/or BCRP and/or MRP1, lymphomas strongly expressed P-gp, weakly expressed BCRP and did not or showed weak expression of MRP1. There was very little staining for MRP2 in the tumours. Newly formed vessels in all tumours only expressed P-gp and BCRP. P-gp, BCRP and MRP1 are present in different cell layers of the normal testis, suggesting the optimal protection of spermatogenesis. In germ cell tumours, this expression pattern may explain the chemoresistance observed to P gp, BCRP and MRP1 substrates. In germ cell tumours and testicular lymphomas, P-gp and BCRP expression by tumour cells and by newly formed vessels may also contribute to chemoresistance. These findings underscore the importance of removing the affected testis in cases of primary germ cell tumours and testicular lymphomas, irrespective of whether the patient has already undergone chemotherapy. PMID- 15341982 TI - Weekly high-dose 5-fluorouracil and folinic acid in metastatic pancreatic carcinoma: a phase II study of the EORTC GastroIntestinal Tract Cancer Cooperative Group. AB - The aim of the study was to assess the response rate and toxicity of high-dose 24 h infusion of 5-fluorouracil (5FU) in metastatic adenocarcinoma of the pancreas. Patients with measurable disease, performance status 0-2, and no prior chemotherapy were registered to receive cycles of leucovorin (LV) 500 mg/m2 (or l LV 250 mg/m2 over 1 h followed by 5FU 2.6 g/m2 over 24 h, weekly for 6 weeks, followed by a 2-week rest. The main endpoints were the response rate and toxicity. From 37 patients, 36 were the analysed for toxicity, and 33 were eligible and analysed for response. The median age was 59 years (range 28-74 years), and the median performance status was 1. Partial response was observed in three patients (9%) (95% Confidential Interval (CI): [2-24]%). Main grade 3/4 National Cancer Institute (NCI) common toxicity criteria toxicities (patients) were diarrhoea (n = 3), vomiting (n = 2) and hand-foot syndrome (n = 5). Median time to progression was 7 weeks (95% CI: [6.4-11.7] weeks) and median survival 19 weeks (95% CI: [12-35] weeks). In conclusion, high-dose 5FU and folinic acid is well tolerated, but has only modest activity in pancreatic cancer. PMID- 15341981 TI - Results of a randomised phase II study comparing docetaxel with methotrexate in patients with recurrent head and neck cancer. AB - We report the results of a randomised phase II trial of docetaxel tested as a single agent in patients with recurrent head and neck cancer using methotrexate as a control arm to validate the results. Eligibility criteria included: histologically-confirmed squamous cell carcinoma, measurable disease, adequate haematological, renal and hepatic functions, no prior chemotherapy for recurrent cancer, signed informed consent. 40 mg/m2 methotrexate was given as a short weekly bolus i.v. injection, and 40 mg/m2 docetaxel was administered as a one hour weekly infusion. A total of 57 patients were randomised based on a ratio of 2/1:37 and 20 patients received docetaxel and methotrexate, respectively. Patient characteristics included 49 males and 8 females; the median age was 59 years (range: 43-82 years). Twenty-eight patients had a local-regional relapse and 29 had distant metastasis, the median disease-free interval was 7.9 months (range: 0 165 months). For patients treated with docetaxel, the following grade 3-4 toxicities occurred: neutropenia (12.5%) with febrile neutropenia in one patient (1%), anaemia (19%) mucositis (9%) and ungueal toxicity (9%). In the methotrexate arm, the grade 3-4 toxicities were: anaemia (15%) and mucositis (5%). The response rate was significantly higher in the docetaxel arm with 27% (95% confidence interval (CI): 21.7-32.3%) of objective responses versus 15% (95% CI: 11.2-18.8%) in the methotrexate arm. Overall survival and time to progression were super-imposable between the docetaxel and methotrexate treatments. Docetaxel given as a weekly infusion has a high activity in patients with head and neck cancer. A phase III trial is needed to test if this translates into a survival benefit for docetaxel use. PMID- 15341983 TI - Central nervous system imaging in childhood leukaemia. AB - The aim of this study was to document the imaging abnormalities seen in the central nervous system (CNS) in cases of childhood leukaemia or as complications of its treatment. Magnetic Resonance (MR) images and Computed Tomographic (CT) scans were reviewed retrospectively in 22 children and adolescents with neurological manifestations/complications of leukaemia or its treatment. Among the 22 patients, nine had two or more different CNS abnormalities. The imaging abnormalities seen in 15 patients before or during treatment included sinus thrombosis, cortical vein thrombosis, cerebral haemorrhage, meningeal leukaemia, infections, skull leukaemic infiltration and treatment-related neurotoxicity. After therapy, seven patients had CNS abnormalities, including secondary brain tumours, skull tumour, mineralising microangiopathy, leucoencephalopathy, transient white matter abnormalities, spinal intradural haematoma, chronic subdural haematoma, radiation necrosis, meningeal leukaemia and leukaemic infiltration at the vertebral body. CNS complications are related to the inherent risk of leukaemia itself, to the treatment method and to the duration of survival. PMID- 15341984 TI - New technology for radiotherapy in paediatric oncology. PMID- 15341985 TI - New technologies in radiation oncology: opportunities, caveats and the way forward. Commentary on "New technology for radiotherapy in paediatric oncology" by Dr. Frank Saran. PMID- 15341986 TI - Constipation, laxative use and risk of colorectal cancer: The Miyagi Cohort Study. AB - The objective of this study was to investigate the association between constipation or laxative use and the risk of colorectal cancer in Japanese men and women. In 1990, we delivered a self-administered questionnaire to 41670 subjects who were 40-64 years old. During the seven years of follow-up, 251 incident cases of colorectal cancer were documented. Constipation was defined as a bowel movement frequency of less than daily. The multivariate relative risk (RR) of colorectal cancer for constipated subjects compared with those with daily bowel movements was 1.35 (95% Confidence Interval: 0.99-1.84). The RR for laxative users over non-users was 1.31 (0.88-1.95), and for frequent users (twice a week or more) it was 2.75 (1.48-5.09). When colorectal cancers were divided into colon cancers or rectal cancers, a significant association was found with colon cancer alone. Our results support the hypothesis that constipation or laxative use increases the risk of colon cancer. PMID- 15341987 TI - Process indicators from ten centres in the Finnish breast cancer screening programme from 1991 to 2000. AB - The aim of this study was to evaluate the quality of the Finnish mammography programme by assessing process indicators from 10 screening centres using data from the first and subsequent screens. We compared these screen-specific indicators with European standards and results from countries with similar screening protocols. Ten Finnish centres invited approximately 1,000,000 women from 1991-2000. Women were mainly 50-64 years old. Mean compliance amongst this age group was 90% at the first and 93% at subsequent screens. The corresponding recall rates were 4.6% and 2.3%, respectively. The average breast cancer detection rates were 0.44% and 0.36%, respectively. The positive predictive values (PPVs) of mammography at the first and subsequent screens were 10% (range 7-20%) and 16% (range 12-31%), and the corresponding benign to malignant (B:M) biopsy ratios were 1:1 (range 0.5-1.8:1) and 0.4:1 (range 0.3-0.8:1). The PPV of mammography increased significantly during the study period, and the average process indicators fulfilled the criteria of the European community for the most part. However, the variation in PPVs was wide, as has been seen for other European mammography programmes, indicating meaningful differences in diagnostic criteria and potential adverse effects. PMID- 15341988 TI - Identification of cytochrome P450 enzymes in human colorectal metastases and the surrounding liver: a proteomic approach. AB - We describe the direct identification of multiple cytochrome P450 (CYP) enzymes in healthy and cancerous tissue. CYPs in human liver colorectal metastases were compared with those in the surrounding liver using a mass spectrometry-based proteomic approach. Nano-scale reversed phase liquid chromatography combined with electrospray ionisation tandem mass spectrometry has been used to identify CYPs with no pre-selection of the proteins anticipated. Fourteen distinct CYP enzymes from the subfamilies 1A, 2A, 2B, 2C, 2D, 2E, 3A, 4A, 4F, 8B and 27A were positively identified; 13 in the liver samples and 12 in the tumour tissue. It was found that three of the colorectal metastases expressed essentially the same drug-metabolising pattern of CYPs as the surrounding liver, whilst three tumour samples from different individuals showed no CYP expression. This was likely in at least one case to be due to low sample mass. The CYP expression profile in an individual tumour is likely to be an important determinant in predicting the outcome of cancer chemotherapy. PMID- 15341989 TI - Adenoviral-mediated gene transduction of the hepatocyte growth factor (HGF) antagonist, NK4, suppresses peritoneal metastases of gastric cancer in nude mice. AB - The competitive inhibitory effects of NK4 (a specific hepatocyte growth factor (HGF)-antagonist) on the interaction between HGF and the c-Met/HGF receptor has been shown in HGF-mediated invasion of some distinct types of human cancer cells. Furthermore, NK4 has inhibitory effects on the angiogenic pathways driven by basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), as well as by HGF. In this study, to evaluate the therapeutic efficacy of adenoviral-mediated NK4 gene treatment, we employed animal models of peritoneal metastasis using two gastric cancer cell lines, the strongly c-Met expressing MKN45 cell line and the weakly c-Met-expressing cell line, TMK1. In both models, the total number and weight of peritoneal tumours per mouse and ascites treated early with AxCANK4 (administered 3 times 2, 7 and 12 days after the tumour inoculation) were significantly reduced compared with those treated with phosphate-buffered solution (PBS) and AxCALacZ (P < 0.05). In Factor-VIII-related antigen-stained sections from peritoneal metastatic tumours, the inhibition of intratumour vessels was observed in tissues from tumours of MKN45 and TMK1 treated with AxCANK4. We also compared the therapeutic effect of early AxCANK4 treatment with that of late treatment (at 7, 12 and 17 days). Peritoneal metastases and ascites treated late with AxCANK4 showed less of an improvement than those treated early with AxCANK4 in both models. In addition, the inhibitory effect of cisplatin (CDDP) on peritoneal metastasis was significantly enhanced by AxCANK4, suggesting that the combination of intraperitoneal (i.p.) chemotherapy with NK4 gene therapy might be effective, even in cases of advanced peritoneal metastasis from gastric cancer. To conclude, these results show clearly that NK4 gene therapy inhibits peritoneal metastases from gastric cancer, regardless of the level of c-Met/HGF receptor expression in the tumour cells, and especially in the early stages of peritoneal metastasis. PMID- 15341990 TI - Aberrant expression, processing and degradation of dystroglycan in squamous cell carcinomas. AB - The alpha- and beta- dystroglycan (DG) proteins are involved in epithelial cell development, formation of the basement membrane and maintenance of tissue integrity. Recently, specific changes in the expression patterns of DGs have been described in some cancers. We studied the expression and localisation of alpha- and beta-DG using Western blotting, immunohistochemistry and reverse transcriptase-polymerase chain reaction analyses in samples of normal oral mucosa, oral squamous cell carcinoma (SCC) and cancer cell lines. The alpha- and beta-DG were localised in the basal layers of normal oral mucosa.However, beta-DG expression in cancer tissues showed evidence of aberrant expression, processing and degradation. alpha-DG was altered in all oral cancer samples and cell lines, despite the persistent presence of DG mRNA in cancer cells. Using matrix metalloproteinase (MMP) inhibitors, we determined that beta-DG degradation in carcinoma cell lines can be mediated by MMPs but this process is highly variable, even in cells from the same cancer type. Considering the multifaceted role of DG in epithelial development, it appears that the role of DG degradation in cancer growth and spread, although currently poorly understood, may be important. PMID- 15341991 TI - Linkage between androgen receptor gene CAG trinucleotide repeat length and testicular germ cell cancer histological type and clinical stage. AB - Sex hormones and/or gonadotropins may play a crucial role in the development of testicular germ cell cancer (TGCC). A direct link between this malignancy and endocrine factors has not been confirmed. We tested whether CAG and GGN repeats of the androgen receptor gene (AR) play a role in the aetiology or pathogenesis of TGCC. Eighty-three TGCC patients and 220 controls were included. Mean CAG or GGN lengths did not differ between the TGCC cases and controls. The proportion of males with CAG lengths above 25, indicative of reduced androgen sensitivity, was significantly lower among patients with pure seminomas and in the combined group of seminomas and mixed tumours compared with non-seminomas and controls. The median CAG length was higher if the tumour was metastasing at diagnosis. This is the first study showing an association between the AR polymorphism and histological type as well as the progression rate of TGCC. PMID- 15341992 TI - Plasma TIMP-1 in patients with colorectal adenomas: a prospective study. AB - Colorectal cancer patients have increased plasma levels of tissue inhibitor of metalloproteinases-1 (TIMP-1). However, it remains unresolved whether colorectal adenomas are associated with increased plasma TIMP-1. Plasma TIMP-1 levels were determined using an immunoassay in 121 patients prospectively enrolled from surveillance colonoscopy programmes. TIMP-1 levels were correlated to various clinicopathological parameters. No significant associations were found between plasma TIMP-1 and size, macro- or microscopic morphology or grade of dysplasia of the adenomas. No significant differences in TIMP-1 levels were found between patients with adenomas (n = 36), hyperplastic polyps (n = 12) or no pathology (n = 68) of the large intestine. However, patients with colonic (n = 3) or rectal (n = 2) adenocarcinomas had significantly increased TIMP-1 levels (P = 0.02). The present study demonstrates that measurement of plasma TIMP-1 cannot be used for the identification of adenomatous or hyperplastic polyps of the large intestine. PMID- 15341993 TI - Black tea polyphenol theaflavins inhibit aromatase activity and attenuate tamoxifen resistance in HER2/neu-transfected human breast cancer cells through tyrosine kinase suppression. AB - The aromatase enzyme, which converts androstenedione to oestrone, regulates the availability of oestrogen to support the growth of hormone-dependent breast tumours. In this study, we investigated the inhibitory effects of black tea polyphenols on aromatase activities. We found that black tea polyphenols, TF-1, TF-2 and TF-3, significantly inhibited rat ovarian and human placental aromatase activities. In addition, using an in vivo model, these black tea polyphenols also inhibited the proliferation induced by 100 nM dehydroepiandrosterone (DHEA) in MCF-7 cells. Transfection of HER2/neu in MCF-7 breast cancer cells appeared to be associated with an increased resistance of the cells to hormonal therapy. Interestingly, unlike the selective oestrogen receptor modulator (SERM) tamoxifen, black tea polyphenols had antiproliferation effects in breast cancer cells with hormonal resistance. The inhibitory effect of black tea polyphenols on hormone-resistant breast cancer cells suppressed the basal receptor tyrosine phosphorylation in HER2/neu-overexpressing MCF-7 cells. These findings suggest the use of black tea polyphenols may be beneficial in the chemoprevention of hormone-dependent breast tumours and represent a possible remedy to overcome hormonal resistance of hormone-independent breast tumours. PMID- 15341994 TI - 5'CpG island hypermethylation and aberrant transcript splicing both contribute to the inactivation of the FHIT gene in resected non-small cell lung cancer. AB - The relative contribution of promoter hypermethylation and aberrant splicing to the inactivation of the fragile histidine triad (FHIT) gene is unclear. Using genetic and epigenetic analyses, the current investigation examines the loss of protein and mRNA expression, and 5'CpG hypermethylation and allelic imbalance of the FHIT gene in a series of 129 non-small cell lung cancer (NSCLC) samples, in parallel with clinicopathological analyses. We found that 50% of NSCLC patients had aberrant protein expression, which was more frequent in squamous cell carcinomas (SQ) (69%) than in adenocarcinomas (AD) (28%) (P < 0.0001). 5'CpG hypermethylation of FHIT was identified in 31% of patients. Abnormally-sized FHIT transcripts were also observed in 24% of patients and were attributed to various exonic deletions, mainly in the region of exons 4-8. Allelic imbalance of the FHIT locus and its correlation with the status of Fhit expression, 5'CpG hypermethylation, and aberrant splicing, indicated that biallelic inactivation of Fhit expression could be induced by 5'CpG hypermethylation of one allele and alternative splicing in the other allele. Moreover, an 83% concordance in the methylation status of FHIT was demonstrated between 12 samples of bronchial precancerous lesions taken before surgery and their matched resected tumours. Our data suggest that FHIT 5'CpG hypermethylation and splicing alterations are both predominant mechanisms involved in the aberrant expression of the FHIT gene, and that FHIT 5'CpG methylation may be potentially used as a supplemental detection marker for NSCLC. PMID- 15341996 TI - Toll-like receptors and other links between innate and acquired alloimmunity. AB - Innate immunity represents the first line of defense against invading pathogens and noxious stimuli. The Toll-like receptors (TLRs) are essential innate immune receptors that alert the immune system to the presence of invading microbes. Emerging evidence shows that TLR signaling is important in allograft rejection. In a murine model, the rejection of minor mismatched allografts cannot occur in the absence of MyD88, an important TLR signal adaptor protein, owing to a defect in dendritic cell maturation, which leads to diminished T-helper cell type 1 immune responses. A recent clinical study also suggests that recipients with a mutant TLR4 genotype manifest reduced lung allograft rejection. Thus, innate immune signaling via TLRs is important for alloimmunity. PMID- 15341997 TI - Antibody, complement and accommodation in ABO-incompatible transplants. AB - Many facets of accommodation have been explored since this process was first observed in ABO-incompatible renal allografts over 17 years ago. Intriguing new pieces of the puzzle have emerged to be fitted into the picture in several places. For example, vascular endothelial cells can be stimulated to secrete substantial amounts of blood group A and B antigens linked to von Willebrand factor; the antibody response to A and B antigens stimulated by ABO-incompatible renal allografts can show epitope spreading; complement can inhibit inflammation through actions of some complement split products, particularly iC3b and C3a; endothelial cells can upregulate various cytoprotective mechanisms; and clinically, new protocols for achieving accommodation have been implemented with improved results. PMID- 15341998 TI - New spectrum of allorecognition pathways: implications for graft rejection and transplantation tolerance. AB - It has long been appreciated that MHC alloantigens can be recognized via two pathways; direct and indirect. The relative contributions of these two pathways to transplant rejection are partially understood. In studies of transplantation tolerance it appears that regulatory T cells (Trs) with indirect allospecificity, particularly the CD4+CD25+ population, play a key role and can regulate responder cells with direct allospecificity for the same alloantigens. One of the conundrums that remains is how helper T and Tr cells with indirect allospecificity regulate T cells with direct allospecificity. At face value, this appears to break the rules of linkage that require interacting T cells to make contact with the same antigen-presenting cell. A third, 'semi-direct' pathway involving MHC exchange may help to resolve this conundrum. Insights into how these pathways interact in transplant immunity and tolerance will assist the pursuit of clinical tolerance. PMID- 15341999 TI - Heterologous immunity and homeostatic proliferation as barriers to tolerance. AB - The different threshold of activation for memory T cells compared to that of naive T cells makes them resistant to immunomodulation, thus representing a barrier to tolerance. Recently it has been demonstrated that homeostatic proliferation and heterologous immunity represent two naturally occurring and distinct processes that can generate memory T cells. Homeostatic proliferation refers to the process by which, in a lymphodeficient host, normal T cells 'spontaneously' proliferate in response to self-MHC-peptide complexes. Heterologous immunity refers to a process in which a response to one or more infectious agents generates effector/memory T cells with cross-reactive specificities. Recent new studies have defined the importance of these processes in transplantation models and implicated strategies to induce transplantation tolerance. PMID- 15342000 TI - Biological effects of induction immunosuppression. AB - The concept of induction immunosuppression is evolving. Once used to buttress the inadequacies of past maintenance immunosuppressive regimens, it is now being used to permit effective maintenance immunosuppression using ever decreasing amounts of modern agents. In addition to lymphocyte depletion, with which it was once synonymous, it is now recognized that induction immunosuppression is associated with a host of non-depletional effects such as receptor modulation and blockade, which profoundly alter the lymphocyte's capacity to mount an effective response. Additionally, the recent focus on the effect of induction agents on antigen presenting cells and on regulatory factors controlling homeostatic repopulation may ultimately permit a safer, more refined and more effective approach to induction immunosuppression. PMID- 15342001 TI - Therapeutic manipulation of T cell chemotaxis in transplantation. AB - T cell migration and trafficking are regulated by the well defined cellular processes of rolling, activation, tight adhesion, arrest and diapedesis. These processes are, in turn, controlled by molecular events involving integrins, selectins, chemokines and chemokine receptors. Recent studies have shown that sphingosine 1-phosphate receptors and their ligands are also important molecular modulators of migration and trafficking. Many of these molecules are appropriate targets for preventing allograft rejection or for achieving tolerance. Studies of migration and trafficking have also shown that the anatomic choreography of alloantigen presentation and T cell encounter with alloantigen and immunosuppression, are over-riding determinants of T cell priming versus tolerization. PMID- 15342002 TI - Chimerism as a tool to induce clinical transplantation tolerance. AB - The principle that the induction of (mixed) hematopoietic chimerism can lead to transplantation tolerance to another organ from the same donor has been verified in rodents, in large animals including non-human primates and recently in a selected group of renal transplant recipients. The wide application of this tool depends on the development of more gentle, non-toxic induction protocols and reliable assays with which to detect the establishment of stable donor-specific tolerance. PMID- 15342004 TI - The complement system in prion diseases. AB - The complement system is an essential component of the innate immune system that aids the recognition and destruction of pathogens. Despite the potent cytolytic activity of this system, some pathogens have turned interactions with complement to their advantage. Transmissible spongiform encephalopathies (TSEs), or prion diseases, also seem to use the complement system to facilitate target cell infection in lymphoid tissues. TSEs are associated with the accumulation of disease-specific prion protein in the brain, which is accompanied by progressive neurodegeneration. Unregulated activation of complement can cause host tissue damage, as observed in some neurodegenerative disorders, and active complement components have been detected in the brains of hosts with TSEs. Whether complement inhibitors will be useful to treat TSEs remains to be determined. PMID- 15342005 TI - Recent developments in prion immunotherapy. AB - Antibody-based immunotherapy may represent a realistic approach against prion diseases, given that antibodies to the cellular prion protein PrPC have been shown to antagonize deposition of the disease-associated prion protein (termed PrPSc) in in vitro assays and in laboratory animals. However, induction of protective antiprion immune responses in wild-type animals is difficult because of host tolerance to the endogenous PrPC. Several studies indicate that it might be possible to overcome tolerance to PrPC and induce immune responses to bacterially expressed, recombinant PrP. However, it is much more difficult to induce antibodies capable of recognizing native cell-surface PrPC, and there is reason to believe that the latter immune responses correlate with anti-prion protection. The difficulties involved in eliciting development of such anti native PrPC immune responses may be partly intrinsic to B cells and, in addition, may reside in peripheral T helper tolerance. PMID- 15342006 TI - Current progress in beta-amyloid immunotherapy. AB - As neuroscientists, we are taught that the brain is immune privileged and thus unlikely to be affected by the peripheral immune system. Accordingly, initial results demonstrating the effectiveness of beta-amyloid (Abeta) immunotherapy in mouse models of Alzheimer's disease (AD) were viewed with considerable surprise and some skepticism. Many groups have since demonstrated efficacy with Abeta immunotherapy in models of AD, using Abeta-based immunogens and anti-Abeta antibodies. Clinical trials involving Abeta immunotherapy for AD are in progress and are providing a wealth of information around the amyloid hypothesis of AD. Abeta immunotherapy is also raising new opportunities and questions about the general role of the immune system in neurodegenerative diseases. PMID- 15342007 TI - Toward modeling hemorrhagic and encephalitic complications of Alzheimer amyloid beta vaccination in nonhuman primates. AB - The potential of amyloid-beta (Abeta) immunization as a disease-modifying therapy for Alzheimer's disease is limited by the occurrence of encephalitic side effects in a subset of treated patients. The encephalitis was not predicted from immunization studies in transgenic, Abeta-depositing mice. More recently, studies in these same mice indicate that passive immunization with certain anti-Abeta antibodies can induce microhemorrhage. Cerebral amyloid angiopathy (CAA) may play a key role in determining the risk for these complications. Because aged nonhuman primates (NHPs) have a more human-like immune system than rodents, and because NHPs naturally develop senile plaques and CAA with age, NHPs appear to be important, adjunctive models for assessing the efficacy and safety of immunotherapeutics for Alzheimer's disease. Conversely, the ability to model the complications of Abeta immunotherapy will be important for elucidating the bases of these complications, and for developing protocols that minimize or eliminate the risks of these serious adverse effects. PMID- 15342008 TI - Neuroimmunology of the paraneoplastic neurological degenerations. AB - The paraneoplastic neurological degenerations (PNDs) are remarkable examples of naturally occurring tumor immunity in humans. In PND patients, common tumors such as breast, ovarian and lung tumors express proteins normally made exclusively in the brain, eliciting an immune response that successfully suppresses growth of the tumor. This successful anti-tumor response would be expected to go clinically unnoticed by the patient, but the immune cells mediating the response somehow cross into the brain, resulting in an autoimmune attack on neurons and neurological symptoms. Pieces of the mystery of this tumor immune response and neuronal autoimmunity have been assembled, but much more needs to be learned. PMID- 15342010 TI - Killer immunoglobulin-like receptors. AB - Killer Ig-like receptors (KIRs) are surface inhibitory receptors specific for allelic forms of human leukocyte antigen (HLA) class I molecules, which are expressed by natural killer (NK) cells and a subset of T lymphocytes. Upon engagement with HLA class I molecules, KIRs block NK cell activation and function. Cells lacking HLA class I molecules are promptly killed by NK cells because of the predominant effect of several activating NK receptors. The NK mediated killing of these cells might represent an important defence mechanism, antagonizing spreading of pathogens and tumours. Evidence has been accumulated that KIR-encoding genes have evolved and diversified rapidly in primates and in humans. Similar to HLA loci, KIR sequences are highly polymorphic and, moreover, KIR haplotypes greatly vary in the number of the type of genes they contain. KIR gene expression is regulated by mechanisms of DNA methylation. As recently shown, the HLA class I regulated control of NK cell function can be exploited in an allogeneic bone marrow transplantation setting to eradicate acute myeloid leukaemias. PMID- 15342011 TI - Inhibitory killer Ig-like receptor genes and human leukocyte antigen class I ligands in haematopoietic stem cell transplantation. AB - Identification of inhibitory killer Ig-like receptor (KIR) genes and their cognate human leukocyte antigen (HLA) class I ligands in donor-recipient pairs for patients undergoing haematopoietic stem cell transplantation (HCT) as treatment for haematopoietic malignancies has recently gained considerable interest. One incentive for these studies is to identify potential donors who are not HLA identical with the recipient but who still could serve as acceptable and potentially preferred donors based on their KIR genotype. It is demonstrated that a majority of individuals have at least one inhibitory KIR gene for which they lack the cognate HLA class I ligand. Therefore, the clinical benefits conferred by 'missing KIR ligand' might not be limited only to HLA mismatched donor recipient combinations but may be applicable also to HLA identical transplants and even autologous HCT. PMID- 15342012 TI - HLA class I antigen expression in malignant cells: why does it not always correlate with CTL-mediated lysis? AB - HLA class I antigen defects are frequently found in malignant cells. They appear to play a role in the clinical course of the disease, probably because they provide tumor cells with a mechanism to escape cytotoxic T lymphocyte (CTL) recognition and destruction. Expression of HLA class I antigens, however, is not always associated with the susceptibility of tumor cells to CTL lysis. Many mechanisms may underlie this finding, including the lack of tumor antigen (TA) derived peptide presentation by a given HLA class I allospecificity, and/or the expression of immunosuppressive molecules such as HLA-G. These findings emphasize the need to develop probes to measure HLA class I allospecificity-TA peptide complex expression in malignant cells. Furthermore, the evaluation of the role of HLA class I antigens in the interaction of malignant cells with host immune cells should take into account the potential interference of tumor-derived immunomodulators. PMID- 15342013 TI - TAP genes and immunity. AB - The transporter associated with antigen processing (TAP) is a member of the ATP binding cassette transporter family that specializes in delivering cytosolic peptides to class I molecules in the endoplasmic reticulum. The TAP is a major target of genetic alteration in tumours and disruption by viral inhibitors. In some species, TAP genes have co-evolved with MHC class I molecules to deliver peptides that are customised for particular alleles. In humans, MHC class I polymorphism determines the level of tapasin-mediated association with TAP and subsequent peptide optimisation within the peptide-loading complex (PLC). MHC class I molecules that still load peptides without complexing to the TAP might be more resistant to viral interference of the PLC and less sensitive to competition for TAP by other class I allotypes. PMID- 15342014 TI - The genetics of HLA-associated disease. AB - Type 1 diabetes mellitus (T1D) remains the most intensively studied, and thus the best paradigm, of MHC-associated diseases. Accumulating evidence suggests that MHC susceptibility for T1D is recessive, with susceptibility alleles more common than protective alleles. Updated allele-level and nucleotide sequence analysis of MHC class II T1D susceptibility markers of conserved extended haplotypes underscore the uncertainty surrounding the actual T1D MHC susceptibility locus. Recent studies have established that disease concordance in dizygotic twins is the same as that in siblings generally, for both T1D and the MHC-associated autoimmune disease gluten-sensitive enteropathy, leaving little room for a differential environmental trigger. Epigenetic mechanisms are probably involved in many MHC-associated phenomena, including autoimmunity, and appear to be the best explanation for incomplete penetrance. PMID- 15342015 TI - Genetics of the central MHC. AB - The MHC, primarily known for its antigen-presenting class I and II molecules, harbours, within a central segment of less than 1 Mb, a dense collection of genes involved in various biological functions. Although MHC I and MHC II are principal players of adaptive immunity, several loci within this central (still called class III) MHC region encode members of the innate immune system. These include the long known factors of the complement system--potentially inhibitory and triggering natural killer receptors as well as stress proteins. Whether this physical proximity is fortuitous or functionally advantageous is an important question for the future of MHC genetics. PMID- 15342016 TI - Effect of surface grafted polymers on the adsorption of different model proteins. AB - Adsorption of a model protein to a surface with end-grafted polymers was studied by Monte Carlo simulations. In the model the effect on protein adsorption in the presence of end-grafted polymers was evaluated by calculating the change in free energy between an end-grafted surface and a surface without polymers. The change in free energy was calculated using statistical mechanical perturbation theory. Apart from ordinary athermal polymer-polymer and protein-polymer interactions we also study a broad selection of systems by varying the interaction between proteins and polymers and effective polymer-solvent interactions. The interactions between the molecules span an interval from -0.5 to +0.5 kT. Consequently, general features of protein adsorption to end-grafted surfaces is investigated by systematically changing properties like hydrophilicity/hydrophobicity of the polymer, protein and surface as well as grafting density, degree of polymerization and protein size. Increasing grafting density as well as degree of polymerization decreases the adsorption of protein except in systems with attractive polymer-protein interactions, where adsorption increases with increasing chain length and higher grafting density. At a critical polymer-protein interaction neither chain length nor grafting density affects the free energy of adsorption. Hydrophilic polymers were found to prevent adsorption better than hydrophobic polymers. Very small particles with radii comparable to the size of a polymer segment were, however, better excluded from the surface when using hydrophobic than hydrophilic polymers. For systems with attractive polymer-protein interaction not only the volume of the protein was shown to be of importance but also the size of the exposed surface. PMID- 15342017 TI - Adsorption kinetics of some carotenoids at the oil/water interface. AB - The kinetic analysis of the adsorption of two carotenoids (i.e., ethyl ester of beta-apo-8'-carotenoic acid and beta-carotene, all trans-isomers) from n-hexane solutions at the oil/water interface is presented for several carotenoid concentrations in the oil phase. A new kinetic approach is developed and it addresses the diffusion adsorption associated with a reversible interfacial reaction, which describes the reorientation of surfactant molecules between two conformations. This approach leads to a general analytical expression that contains four physical parameters and describes with high accuracy the experimental dynamic interfacial tensions for the two carotenoids, which independently adsorb from n-hexane phase to the n-hexane/water interface. The calculations give the characteristic times for the carotenoid adsorption at the oil/water interface in terms of diffusion relaxation and kinetic relaxation times. The results explain the long time effects on the adsorption of these carotenoids at the oil/water interface. The data are in substantial agreement with the molecular structure of these carotenoids and with the earlier data recorded for cholesterol adsorption at the n-heptane/water interface. Based on these findings, we propose a molecular mechanism for the interfacial transformation of carotenoid molecules at a hydrophobic/hydrophilic interface. PMID- 15342018 TI - Selective separation of pyrite and chalcopyrite by biomodulation. AB - Selective separation of pyrite from other associated ferrous sulphides at acidic and neutral pH has been a challenging problem. This paper discusses the utility of Acidithiobacillus ferrooxidans for the selective flotation of chalcopyrite from pyrite. Consequent to interaction with bacterial cells, pyrite remained depressed even in the presence of potassium isopropyl xanthate collector while chalcopyrite exhibited significant flotability. However, when the minerals were conditioned together, the selectivity achieved was poor due to the activation of pyrite surface by the copper ions in solution. The selectivity was improved when the sequence of conditioning with bacterial cells and collector was reversed, since the bacterial cells were able to depress collector interacted pyrite effectively, while having negligible effect on chalcopyrite. The observed behaviour is analysed and discussed in detail. The separation obtained was significant both at acidic and alkaline pH. This selectivity achieved was retained when the minerals were interacted with both bacterial cells and collector simultaneously. PMID- 15342019 TI - Effect of chemical and physical factors to improve the germination rate of Echinacea angustifolia seeds. AB - Seeds of Echinacea angustifolia are known for their deep dormancy. In this paper, we studied the responses of E. angustifolia seeds to some chemical and physical factors, such as scarification, chilling (5 degrees C) period, light and applied BA (6-benzylaminopurine), GA3 (gibberellic acid) and sound stimulation. When the seed coat layers were removed, the germination rate grew up from 6 to 20% (incubated in light) and the mean time germination (MTG) was reduced from 18 to 6.6 days. On the basis of layers-removed, chilling and continuous light gave significantly higher germination rate (up to 70%). Compare the data of seeds chilled by 0, 6, 12, 18 and 24 days, the maximum germination rate (up to 70%) achieved at 18-days chilling treatment. Further increases in the chilling period could slightly improve germination. Exogenous application of 0.1, 0.2, 0.3 mg/L GA3 or BA in the previous pretreatment increased germination to 78, 90 and 84% or 76, 86 and 84%, respectively. Obviously, the best concentration of GA or BA is 0.3 mg/L. And the GA3 or BA treatment shortened the MTG to about 4 days. The influence of sound stimulation was also tested in the experiment. The result showed that one 100 dB and 1000 Hz sound wave (sine-wave) was beneficial to the germination of E. angustifolia seeds. PMID- 15342020 TI - Soundwave stimulation triggers the content change of the endogenous hormone of the Chrysanthemum mature callus. AB - Hormones have been suggested to play a prominent role in the control of callus growth. In this paper, with the method of indirect enzyme-linked immunosorbent assays (ELISA), we investigated the induction effect of soundwave on the dynamic change of endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) in vitro during the differentiation process of Chrysanthemum synchronized mature Callus. These experiments showed that groups treated by optimal soundwave (1.4 kHz, 0.095 kdb) had significantly higher IAA levels and lower ABA than that of the control, which had been implicated activation of endogenous IAA and inhibition of ABA. Through the biochemical analysis, it revealed that the increased level of IAA as well as decreased levels of ABA correlated with soundwave stimulus. High rate of IAA/ABA was favorable to development of the callus and differentiation of mature callus. We conclude that soundwave contributes to endogenous hormone as well as the control of callus growth. PMID- 15342021 TI - Effects of temperature and pH on the helicity of a peptide adsorbed to colloidal silica. AB - The conformation of a cationic alpha-helical peptide (DDDDAAAARRRRR) adsorbed to anionic colloidal silica has been investigated by circular dichroism (CD) spectroscopy as a function of temperature and pH in order to examine how the structure of an adsorbed molecule responds to two simultaneous perturbations. Increased temperature destabilizes the helicity of the peptide in solution, while pH changes alter the substrate surface charge and the corresponding strength of the interaction with the peptide. Near neutral pH, the helicity of the adsorbed peptide, which is determined from the intensity of the CD signal at 222 nm, decreases with increasing temperature, similarly to the temperature-dependent behavior observed for the peptide in aqueous solution. By contrast, at basic pH and a strongly negative surface charge, the helicity of the adsorbed peptide increases with temperature. In order to elucidate the origin of the reversal of the temperature dependence of helicity, a statistical model for the conformation of the adsorbed peptide has been formulated based on the Lifson-Roig model for the helix-coil transition of the peptide in solution. The model provides insight into the trends in fractional helicity and reveals that the temperature dependence of the helicity of the adsorbed peptide results from a competition between the intramolecular interactions that promote helicity and the intermolecular interactions with the surface. The statistical model also enables estimation of the free energy contributions from specific aspects of the adsorption process. Through identification of a connection between the conformation of adsorbed peptide and the interactions of the peptide with the surface, this work suggests a route for the control of adsorbate conformation through peptide and surface engineering. PMID- 15342022 TI - Single-particle light scattering study of polyethyleneglycol-grafted poly(ureaurethane) microcapsule in ethanol. AB - Microcapsules having polyethyleneglycol-grafted poly(ureaurethane) (PUU) membrane and di-2ethylhexyl phthalate core have been prepared, and the structure when they were suspended in dispersing ethanol have been studied by means of single particle light scattering method. The PUU membrane was synthesized from monomers with aromatic functional groups (microcapsule MC110) and hexamethylene functional groups (microcapsule MC160). Because the outer and inner solvent passed through the membrane easily, the outside and inside of the membrane were the same at the equilibrium state. The thickness of the wall membrane was significantly smaller than that calculated from the overall weight ratio of the wall-forming material and the core solvents. It was attributed to low affinity of PUU membranes and ethanol. PMID- 15342023 TI - Characterization of the surface properties of persimmon leaves by FT-Raman spectroscopy and wicking technique. AB - To fit the request of developing new biomaterials using persimmon leaves as raw material, this paper reported the surface free energy and related components, e.g. the Lifshitz-van der Waals and Lewis acid-base components for persimmon leaves estimated by means of the column wicking technique. Considering this natural material might be varied of its properties with growing area, a characterization of it was initially performed by applying the FT-Raman spectroscopy. Based on the determined results, persimmon leaves have greater surface free energy, gammaS, than cellulose though it has been found rich in this component, e.g. of about 68.28%. Moreover, it was further observed that the persimmon leaves have great Lewis acid-base interactions component, gammaS(AB), contributed by the Lewis acid component, gammaS+. This suggests that the persimmon leaves are available to convert to new biomaterials expected forms. PMID- 15342024 TI - Validating and disproving what we "know". PMID- 15342025 TI - Environmental factors and LASIK. PMID- 15342027 TI - Single-piece acrylic intraocular lens implantation in children. PMID- 15342029 TI - Incomplete flap with characteristics similar to corneal ectasia. PMID- 15342031 TI - Incomplete flap with characteristics similar to corneal ectasia. PMID- 15342032 TI - Preoperative laser in situ keratomileusis screening: an evolving standard of care. PMID- 15342033 TI - LASIK advertising: we should not sell procedures. PMID- 15342034 TI - Iridoschisis associated with lens subluxation. PMID- 15342035 TI - Cataract surgery and intraocular lens implantation in retinoblastoma. PMID- 15342038 TI - Consultation section: refractive surgical problem. PMID- 15342042 TI - Removal of a capsular tension ring from the anterior chamber angle. AB - A 75-year-old woman presented with unexplained ocular hypertension 4 weeks after phacoemulsification. Steroid response was diagnosed initially until a misplaced capsular tension ring (CTR) was identified in the drainage angle. We describe the technique used to successfully remove the CTR from the drainage angle with minimal trauma. This case illustrates the importance of the initial surgical technique used for CTR insertion and emphasizes the role of gonioscopy in such a case. PMID- 15342043 TI - Chopstick technique for nucleus removal in an impending dropped nucleus. AB - We describe a bimanual chopstick technique for nucleus removal after a posterior capsule tear and an impending dropped nucleus during phacoemulsification. The technique stabilizes the nucleus and nuclear fragments by providing posterior support with a Sinskey hook introduced from the pars plana. Once supported, the nucleus is gripped between 2 instruments, brought out of the capsular bag into the anterior chamber, and then taken out of the enlarged wound. This bimanual removal technique causes minimal disturbance to the vitreous, iris, and cornea. PMID- 15342044 TI - Intraoperative peripheral anterior capsulotomy to prevent early postoperative capsular block syndrome. AB - We describe a new surgical technique, intraoperative peripheral anterior capsulotomy (IPAC), to prevent early postoperative capsular block syndrome (CBS). Eighty consecutive patients (80 eyes) had phacoemulsification and intraocular lens implantation with IPAC in the presence of a small (4.5 to 5.0 mm) anterior continuous curvilinear capsulorhexis (CCC). Patients were followed for a mean of 16 months (range 6 to 24 months), and none developed CBS or other complications. Our series suggests that IPAC may be an alternative to prevent CBS in high-risk eyes, including those with a small anterior CCC or ophthalmic viscosurgical device material trapped in the capsular bag. This surgical technique is safe, effective, inexpensive, and easy to perform. PMID- 15342045 TI - Static and dynamic analysis of the anterior segment with optical coherence tomography. AB - PURPOSE: To study biometric modifications of the anterior segment with accommodation and age and determine possible applications in areas of anterior segment surgery, particularly implantation of refractive lenses. SETTING: Clinique Monticelli, Marseille, France. METHOD: The study comprised subjects between 7 years of age and 82 years of age in whom anterior chamber biometry was evaluated using 1,310 nm wavelength optical coherence tomography (OCT). The equipment has a fixation target that can be focused and defocused with negative lenses to stimulate natural accommodation. All measurements were performed by the same operator. The horizontal diameter of the AC, the anterior chamber depth (ACD), the horizontal pupil diameter, and the horizontal radius of curvature of the crystalline lens' anterior pole were measured in the unaccommodated state and after stimulating accommodation. RESULTS: Fifty-six subjects (104 eyes) were included; the refractions ranged from +5.0 diopters (D) to -5.0 D. The static and dynamic measurements were compared with ametropia, age, and accommodation. At rest, the mean AC diameter was 12.334 mm, the mean ACD was 3.106 mm, and the mean pupil diameter was 4.258 mm. With 1.0 D of accommodation, the anterior pole moved forward by a mean of 30 microm, the radius of curvature decreased 0.3 mm, and the pupil diameter decreased 0.15 mm. CONCLUSIONS: The AC OCT is a user-friendly instrument for evaluating the anterior segment and examining the AC (cornea, iris, crystalline lens, and iridocorneal angle). The 1,310 nm light wavelength is blocked by pigments, preventing examination behind the iris. However, the AC OCT is capable of good image quality and visualization of the anatomical relationships in the anterior segment, even behind an opaque cornea. PMID- 15342046 TI - Monitoring corneal structures with slitlamp-adapted optical coherence tomography in laser in situ keratomileusis. AB - PURPOSE: To monitor corneal structures with slitlamp-adapted optical coherence tomography (OCT) in laser in situ keratomileusis (LASIK). SETTING: Department of Ophthalmology, Vivantes Klinikum Neukolln, Berlin, Germany. METHODS: In this prospective, nonrandomized, comparative clinical case series of consecutive patients who had LASIK for myopia and myopic astigmatism, the corneal structures were studied with slitlamp-adapted OCT at a wavelength of 1,310 nm. The central corneal thickness (CCT) and epithelial, flap, and residual stromal thicknesses were assessed preoperatively, immediately after surgery, on postoperative day 1, and then, on average, after 8, 35, and 160 days. RESULTS: Twenty-five eyes of 13 patients were included. The attempted mean spherical equivalent correction was 6.11 diopters (D) +/- 2.16 (SD) with a mean calculated stromal ablation depth of 92 +/- 24 microm. The CCT was 516 +/- 26 microm preoperatively and 453 +/- 40 microm postoperatively (P<.001). The epithelial thickness increased from 57.0 +/- 7.7 microm preoperatively to 61.0 +/- 7.5 microm postoperatively (P =.04). Imaging of the hyperreflective interface was possible in all patients for up to 15 months. The flap and residual stromal thickness was 211 +/- 28 microm and 344 +/- 48 microm, respectively, immediately after LASIK and 164 +/- 21 microm (P<.001) and 284 +/- 32 microm (P<.001), respectively, on postoperative day 1. There were no further significant changes during the follow-up. The overall mean reproducibility was +/-4.50 microm (coefficient of variation [CV] 0.94%) for CCT, +/-4.99 microm (CV 8.57%) for epithelial thickness, +/-6.25 microm (CV 3.55%) for flap thickness, and +/-7.09 microm (CV 2.42%) for residual stromal thickness. CONCLUSION: Slitlamp-adapted OCT can be used to longitudinally monitor the variable structures of the cornea, epithelium, flap, and residual stroma in LASIK. PMID- 15342047 TI - Reasons patients recommend laser in situ keratomileusis. AB - PURPOSE: To evaluate the reasons patients who have had laser in situ keratomileusis (LASIK) recommend it to others and examine the disparity between high levels of satisfaction and patient reports of night-vision symptoms and/or dry eye after LASIK. SETTING: Northeastern Eye Institute, Scranton, and the University of Pittsburgh, Pittsburgh, Pennsylvania, and the University of California at Los Angeles, Los Angeles, California, USA. METHODS: Questionnaires assessing symptoms and satisfaction after LASIK were mailed to 2,100 patients. The questionnaires included items about night-vision symptoms and satisfaction and an open-ended question for patients to give their reasons for recommending LASIK to others. The open-ended responses were categorized and tabulated. RESULTS: Four hundred thirty-four patients provided reasons for recommending LASIK to others. Sixteen categories of reasons were identified. "No more spectacles/contact lenses" was listed by 180 patients (42%), followed by "better vision" (21%) and "convenience" (15%). Women were significantly more likely to cite "better comfort" (27 women versus 3 men; chi square = 8.99, P =.003) and "better quality of life" (41 women versus 9 men; chi square = 7.36, P =.007) as a reason for recommending LASIK. Of the 35 patients who reported dissatisfaction with post-LASIK vision, 20 (57%) would recommend LASIK to a friend because "LASIK helps others." CONCLUSIONS: Categories of reasons for recommending LASIK to others were similar to reasons given by patients for seeking LASIK. Some patients who reported dissatisfaction with their vision said they would recommend LASIK, suggesting that recommendation of LASIK to others is not necessarily a measurement of the quality of a patient's vision after LASIK. PMID- 15342048 TI - Survey of complications and recommendations for management in dissatisfied patients seeking a consultation after refractive surgery. AB - PURPOSE: To review complications associated with and management options for dissatisfied patients seeking a consultation after refractive surgery performed elsewhere. SETTING: Refractive Eye Surgery Center, The Wilmer Institute, Lutherville, Maryland, USA. METHODS: In this retrospective review, charts of consecutive dissatisfied patients who sought a refractive consultation between June 1999 and January 2003 after refractive surgery performed elsewhere were reviewed and the following parameters were analyzed: visual acuity (uncorrected and best spectacle corrected), prior manifest refraction, complications, and recommendations. The subjective complaints, complications, and visual acuity were reviewed, and the associated historical or visual risk factors as well as treatment options were analyzed. RESULTS: One hundred sixty-one eyes in 101 patients with ocular complaints were identified. One hundred thirty-four eyes (83.2%) had had laser in situ keratomileusis; 22 eyes, photorefractive keratectomy; 4 eyes, radial keratotomy; and 1 eye, laser thermokeratoplasty. The most common subjective complaints were blurred distance vision (59.0%), glare and night-vision disturbances (43.5%), and dry eyes (21.1%). The most common complications were overcorrection (30.4%), irregular astigmatism (29.8%), dry eyes (29.8%), glare (26.1%), difficulty with night driving (16.7%), and corneal haze (16.7%). The most common cause identified in eyes with best spectacle corrected visual acuity worse than 20/40 was irregular astigmatism (10 of 18 eyes [55.5%]). The most common recommendation for management was medical treatment or observation (68.3%). Medical therapies recommended included lubrication, punctal plugs, topical and systemic pharmaceutical agents to modulate ocular surface, spectacles, and contact lenses. Keratoplasty (lamellar or penetrating) was recommended in 4 patients (4 eyes [2.5%]). The diagnoses included corneal ectasia in 3 eyes and severe flap complication with irregular astigmatism in 1 eye. Nine patients (5.6% eyes) required nonkeratoplasty surgery. In other patients, waiting for advances in technology, including wavefront-guided customized retreatment, was recommended. CONCLUSIONS: A spectrum of complications associated with refractive surgery may result in patient dissatisfaction. Proper patient selection, prevention strategies, and prompt diagnosis and medical or surgical intervention may be beneficial in managing complications and improving patient satisfaction. PMID- 15342049 TI - Effect of intraocular lens optic edge design and material on fibrotic capsule opacification and capsulorhexis contraction. AB - PURPOSE: To examine the influence of intraocular lens (IOL) optic edge design and optic material on fibrosis of the anterior and peripheral posterior capsules and on capsulorhexis contraction. SETTING: Department of Ophthalmology, Medical University of Vienna, Vienna, Austria. METHODS: This randomized controlled patient- and examiner-masked study comprised 210 eyes of 105 patients with bilateral age-related cataract. In Group 1 (n = 53), the Sensar OptiEdge AR40e hydrophobic acrylic IOL with a sharp posterior optic edge was compared with the AR40 acrylic IOL with a round edge. In Group 2 (n = 52), the ClariFlex OptiEdge silicone IOL with a sharp posterior optic edge was compared with the PhacoFlex SI 40 silicone IOL with a round edge All IOLs were manufactured by Advanced Medical Optics, Inc. Standardized digital slitlamp images of anterior capsule opacification (ACO) and fibrotic posterior capsule opacification (PCO) were taken 1 year postoperatively, and digital retroillumination images were taken at 1 week and 1 year. The intensity of fibrotic PCO was graded subjectively (score 0 to 4), ACO was graded objectively (score 0% to 100%), and the capsulorhexis area (mm(2)) was determined objectively. RESULTS: One year after surgery, the mean ACO score was 32% in eyes with the sharp-edged acrylic IOL and 29% in eyes with the round edged acrylic IOL (P<.05). In the silicone group, the mean was 31% and 26%, respectively (P<.05). The mean fibrotic PCO score was lower in eyes with a sharp edged acrylic IOL than in eyes with a round-edged acrylic IOL (0.26 and 0.93, respectively; P<.05) and in eyes with a sharp-edged silicone IOL than in eyes with a round-edged silicone IOL (0.24 and 0.82, respectively; P<.001). At 1 year, the mean capsulorhexis area was statistically significantly smaller in eyes with a sharp-edged silicone IOL than in eyes with a round-edged silicone IOL (P<.05). CONCLUSIONS: Acrylic and silicone IOLs with the sharp OptiEdge design led to significantly less fibrotic PCO but more ACO than round-edged acrylic and silicone IOLs. The sharp-edged silicone IOL caused significantly more capsulorhexis contraction than the round-edged silicone IOL and both acrylic IOLs. PMID- 15342050 TI - Functional vision with cataracts of different morphologies: comparative study. AB - PURPOSE: To evaluate the influence of cataract morphology on the functional vision of patients with age-related cataract and normal macular function and compare subjectively perceived functional impairments to distance visual acuity, reading acuity, and maximum reading speed between cataract types. SETTING: Department of Ophthalmology, Medical University of Vienna, Vienna, Austria. METHODS: Seventy-five patients awaiting first-eye cataract surgery were asked to characterize their visual difficulties in everyday life using a German version of the VF-14 questionnaire. Cataracts were categorized and graded using the Lens Opacities Classification System III. Monocular and binocular visual acuity and reading performance were determined in randomized order. RESULTS: There were significant differences in functional vision between nuclear cataracts and posterior subcapsular cataracts (PSC); the VF-14 score and the self-reported visual satisfaction were significantly lower in PSC patients (P<.05). Posterior subcapsular cataracts significantly increased self-reported impairment in distance and near vision, although the visual acuity was comparable to that in the other groups (P =.9). Significant differences in functional vision were also seen between PSC and nuclear-cortical cataracts (P<.05). No significant differences in functional vision were found between nuclear cataracts and nuclear cortical cataracts (P>.05). CONCLUSIONS: The VF-14 questionnaire reliably evaluated functional differences caused by different cataract morphologies; these differences were underestimated when only visual acuity was measured. Patients with PSC had increased functional impairment, indicating that cataract surgical intervention is indicated at an earlier stage in these patients. The significant differences between the morphological types of cataract should be taken into consideration when the benefit of cataract surgery is to be measured on the basis of functional improvement. PMID- 15342051 TI - Intacs after laser in situ keratomileusis and photorefractive keratectomy. AB - PURPOSE: To evaluate the efficacy of intrastromal corneal ring segments (Intacs, Addition Technology) for the correction of residual myopia after laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK). METHODS: In this prospective noncomparative case series, Intacs were implanted in 5 eyes (4 patients) that developed myopic regression after LASIK or PRK. Intacs were implanted a minimum of 11 months after the laser procedure. The technique was identical to the standard procedure for myopic unoperated eyes. The Intacs thickness was based on the amount of residual myopia. RESULTS: Postoperative results revealed a reduction in the residual myopia and the mean K-reading. No eye lost best corrected Snellen visual acuity. No intraoperative or postoperative complications occurred. CONCLUSIONS: Implantation of Intacs in eyes with myopic regression after LASIK and PRK resulted in a good refractive outcome and an improvement in uncorrected visual acuity. PMID- 15342052 TI - Amoils epithelial scrubber to treat recurrent corneal erosions. AB - PURPOSE: To evaluate the treatment of recalcitrant recurrent corneal erosions (RCE) with superficial keratectomy using the Amoils epithelial scrubber (AES). SETTING: Muncie Eye Center, Muncie, Indiana, USA. METHODS: This study was a retrospective consecutive case series comprising 23 patients (26 eyes) who had superficial keratectomy for RCE using the AES between September 1997 and May 2003 at a private tertiary-care center. The preoperative and postoperative best corrected visual acuities (BCVAs), slitlamp findings, and postoperative incidence of RCE were studied. RESULTS: During a mean telephone survey follow-up of 21.2 months +/- 20.5 (SD) (range 1.4 to 71.3 months), 20 patients (23 eyes, 88%) reported no further RCE symptoms. The BCVA improved or stayed the same in 25 eyes (96%). No significant complications were found. CONCLUSION: Superficial keratectomy using the AES appears to be safe and effective for treating recalcitrant RCE. PMID- 15342053 TI - Corneal flap thickness in laser in situ keratomileusis using the Moria M2 microkeratome. AB - PURPOSE: To determine the predictability of flap thickness in laser in situ keratomileusis (LASIK) using the Moria M2 microkeratome and identify factors that may be related to variations in flap thickness. SETTING: Laser Vision Correction Center, Bascom Palmer Eye Institute, University of Miami School of Medicine, Miami, Florida, USA. METHODS: Charts of 208 patients having same-day bilateral LASIK using the Moria M2 microkeratome were reviewed. Intraoperative pachymetry was performed routinely. The right eye was always treated first. The same suction ring, stop, microkeratome head (110 microm or 130 microm), and blade were used in fellow eyes. Subtraction pachymetry was used to calculate flap thickness. Other collected data included age, keratometry, corneal diameter, and preoperative spherical equivalent (SE). RESULTS: With the 110 microm head and slow translation velocity in both eyes, the mean flap thickness was 151.6 microm +/- 24.0 (SD) and 148.5 +/- 24.3 microm in the right and left eyes, respectively. With the 110 microm head and fast translation velocity in both eyes, the mean thickness was 136.2 +/- 25.5 microm and 132.8 +/- 23.5 microm, respectively. With the 130 microm head and fast translation velocity, the mean flap thickness was 145.8 +/- 25.4 microm and 139.9 +/- 25.5 microm, respectively. Flaps were thinner with fast translation velocity, the 110 microm head, and presumably duller blades used in the left eyes. There was a weak but statistically significant inverse correlation between flap thickness and age and between flap thickness and SE. A stronger correlation was found in flap thickness between right and left eyes. CONCLUSIONS: Flap thickness with the Moria M2 microkeratome was variable. Fast translation velocity, a used (presumably duller) blade, and the 110 microm head produced thinner flaps. Given the potential variation in flap thickness (SD 23.5 to 25.5 microm), intraoperative pachymetry might be an adjunctive measure to prevent residual stromal beds that are thinner than planned, especially in patients with high myopia and/or thin corneas. PMID- 15342054 TI - Phototherapeutic keratectomy in children: 5-year results. AB - PURPOSE: To evaluate the efficacy and safety of phototherapeutic keratectomy (PTK) for the treatment of superficial corneal opacities, surface irregularities, epithelial instability, and reepithelialization failure in pediatric patients and study the visual and refractive changes after combined PTK and photorefractive keratectomy (PRK). SETTING: Department of Ophthalmology, Masaryk University Hospital, Brno, Czech Republic. METHODS: This retrospective clinical study comprised children who had PTK or PTK combined with PRK from September 1996 to January 2000. The goals of treatment were to improve visual acuity and reduce or eliminate subjective ocular discomfort (eg, pain, lacrimation, and photophobia). A Nidek EC-5000 excimer laser was used in PTK mode with a 3.0 to 6.0 mm optical zone and a 4.0 to 7.5 mm transition zone. RESULTS: Forty-one pediatric patients (41 eyes) were included. Twenty-three eyes had PTK only, and 18 eyes had PTK combined with PRK to reduce preoperative myopia (11 eyes) or hyperopia (7 eyes). The mean patient age was 11.4 years (range 8 to 18 years) and the mean follow-up, 4.8 years (range 3 to 6 years). The best spectacle-corrected visual acuity (BSCVA) improved in all patients, and episodes of ocular pain or discomfort, lacrimation, and photophobia diminished. The mean preoperative BSCVA of 6/38 (range 6/10 to 1/60) improved to 6/12 (range 6/6 to 6/38) at the last postoperative examination. Eight eyes gained 5 or more Snellen lines of BSCVA; 11 gained 4 lines, 9 gained 3 lines, 7 gained 2 lines, 5 gained 1 line, and 1 eye was unchanged. No eye lost a line of BSCVA. The mean preoperative spherical equivalent (SE) decreased from -5.32 to -1.16 diopters (D) in the 11 myopic eyes and from +4.72 to +1.51 D in the 7 hyperopic eyes within 3 years of the combined procedure. CONCLUSIONS: Phototerapeutic keratectomy is an effective and safe procedure for the treatment of various surface corneal disorders in children. It can improve best corrected visual acuity and eliminate ocular pain and irritation. Preoperative myopia and hyperopia were effectively reduced by a combination of PTK and PRK. PMID- 15342055 TI - Early uncorrected visual acuity as a measurement of the visual outcomes of contemporary cataract surgery. AB - PURPOSE: To determine the uncorrected visual acuity (UCVA) on the first postoperative day and the fifth week after routine slow-motion phacoemulsification with posterior chamber intraocular lens (IOL) implantation. SETTING: Cincinnati Eye Institute, Cincinnati, Ohio, USA. METHODS: This retrospective chart review performed by 3 research fellows analyzed the UCVA 1 day and 5 weeks postoperatively in 100 consecutive best-case scenario eyes of 99 patients who had routine slow-motion phacoemulsification with implantation of an AcrySof single-piece IOL (Alcon). Reasons for UCVAs worse than 20/40 were sought. The stability of the visual result was analyzed. RESULTS: The UCVA was 20/40 or better in 98% of eyes at 1 day. Ninety-seven percent had a UCVA of at least 20/40 by 5 weeks, confirming stability of acuity. The percentage of patients with a UCVA of 20/20 or 20/25 increased from 49% at 1 day to 77% at 5 weeks. CONCLUSIONS: The UCVA 1 day after slow-motion phacoemulsification was 20/40 or better in 98% of eyes and remained stable to the last measurement at 5 weeks. Early UCVA can serve as a more sensitive measurement than best corrected visual acuity for assessing new surgical technologies and techniques. PMID- 15342056 TI - Autorefraction as an outcome measure of laser in situ keratomileusis. AB - PURPOSE: To determine the limits of agreement between subjective refraction and autorefraction before and after laser in situ keratomileusis (LASIK) to assess whether autorefraction is a valid refractive outcome measure of refractive surgery. SETTING: Ultralase, Leeds, United Kingdom. METHOD: The prospective study involved consecutive preoperative normal patients and post-LASIK patients who had autorefraction using the Nidek ARK 700A autorefractor and careful subjective refraction (masked to autorefraction). Inclusion criteria were age greater than 18 years and healthy eyes with a visual acuity better than 0.1 logMAR (6/7.5) with or without previous LASIK. Refractions were compared by spherical equivalent (SE) using Bland-Altman limits of agreement and astigmatic vector difference using the median and the 95th percentile. The effect of time after treatment and treatment strength were explored. RESULTS: Data were collected from 208 preoperative patients and 237 post-LASIK patients. Preoperatively, the agreement between subjective refraction and autorefraction for the SE was -0.10 diopter (D) +/- 0.35 (SD) and the median difference for the astigmatic vector was 0.28 D with a 95th percentile of 0.72 D. Post-LASIK, the SE agreement was similar, -0.09 +/- 0.39 D, but the astigmatic vector agreement decreased slightly with a median of 0.31 D and a 95th percentile of 1.02 D. This decrease reflected poorer agreement in patients whose pre-LASIK refractive error was greater than +4.00 D. Removing this group brought the median astigmatic difference post-LASIK to 0.27 D with a 95th percentile of 0.87 D, similar to that in the preoperative normals. The percentage within +/-0.50 D and +/-1.00 D of the attempted correction was 56.1% and 78.5%, respectively, with subjective refraction and 51.2% and 78.1%, respectively, with autorefraction. CONCLUSIONS: Autorefraction showed excellent agreement with subjective refraction and was unaffected by refractive surgery except after LASIK for high hyperopia. Most outcomes were correctly classified in the standard categories (+/-0.50 D, +/-1.00 D), illustrating that autorefraction is a valid outcome measure of refractive surgery. PMID- 15342057 TI - Comparison of wavefront aberration changes in the anterior corneal surface after laser-assisted subepithelial keratectomy and laser in situ keratomileusis: preliminary study. AB - PURPOSE: To compare changes in anterior corneal wavefront aberrations after myopic laser-assisted subepithelial keratectomy (LASEK) and laser in situ keratomileusis (LASIK). SETTING: Institute of Ophthalmology, Catholic University, Rome, Italy. METHODS: This prospective study included 36 eyes of 25 myopic patients: 18 eyes of 12 patients had LASEK and 18 eyes of 13 patients, LASIK. The topography data (CSO EyeMap, version 6.2) were used to calculate corneal aberrations with 3.0 mm and 7.0 mm pupils before and 3 months after surgery. RESULTS: Total corneal aberrations increased similarly after LASEK and LASIK with the 7.0 mm pupil but did not change with the 3.0 mm pupil. Coma-like and spherical-like aberrations changed similarly after both procedures, but spherical like aberrations increased after LASEK with the 3.0 mm pupil (P<.05, independent t test). With the 7.0 mm pupil, the amount of achieved correction was positively correlated with changes in total corneal aberrations after LASIK (P =.007) and with spherical-like aberrations after LASEK (P =.03) and LASIK (P<.003). Although there was no significant difference between LASEK and LASIK (P>.05, independent t test), in individual eyes with an achieved correction less than 7.50 diopters (D), spherical-like aberrations increased more after LASEK than after LASIK. CONCLUSIONS: In this preliminary study, myopic LASEK and LASIK changed total and higher-order corneal aberrations. In both procedures, changes in spherical-like aberrations were dependent on the achieved correction. However, in individual eyes, spherical-like aberrations increased more after LASEK than after LASIK for low-moderate achieved correction, suggesting that these procedures may induce the same optical changes in the anterior corneal surface in different ways. PMID- 15342059 TI - Risk factors for failure of simultaneous penetrating keratoplasty and cataract extraction. AB - OBJECTIVE: To evaluate the indications, complications, and outcomes of simultaneous cataract extraction and penetrating keratoplasty (PKP) and the risk factors for failure of the procedure. SETTING: Tertiary referral medical center. METHODS: In a retrospective noncomparative interventional case series, the charts of 66 consecutive patients (70 eyes) who had simultaneous PKP and cataract extraction using the same surgical technique were reviewed. RESULTS: The mean follow-up was 32.4 months (range 6 to 125 months). The primary corneal graft remained clear in 48 eyes (69%) and failed in 22 eyes (31%). Sixteen eyes with a failed graft (73%) had 1 or 2 repeat keratoplasties; 8 (50%) were successful. At the end of follow-up, 56 eyes (80%) had a clear corneal graft. Nine eyes (41%) with a failed primary graft and 2 eyes (4%) with a clear primary graft had intracapsular cataract extraction (P<.001); 13 eyes (59%) and 46 eyes (96%), respectively, had extracapsular cataract extraction (P<.001). All eyes with a failed primary graft and 18 eyes (37%) with a clear primary graft had postoperative complications (P<.001). Eyes with a failed graft had more postoperative surgical interventions (P<.001). There were no statistical differences between eyes with clear grafts and eyes with failed grafts in sex, age, indications for surgery, corneal graft diameter, intraoperative vitreous loss, and intraocular lens placement. CONCLUSIONS: Intracapsular cataract extraction, postoperative complications, and postoperative surgical interventions may increase the risk for graft failure in simultaneous cataract extraction and PKP. Intraocular lens implantation did not increase the risk for graft failure. PMID- 15342058 TI - Opacification of hydrophilic MemoryLens U940A intraocular lenses: analysis of 2 explanted lenses. AB - PURPOSE: To determine the rate of opacification of hydrophilic MemoryLens U940A intraocular lenses (IOLs) (Mentor Ophthalmics, Inc.) in the given cohort and perform a histopathological and spectrophotometer analysis of 2 explanted opacified IOLs. SETTING: Ophthalmology Department, Faculty Hospital, Nitra, Slovakia. METHOD: This retrospective study comprised 182 patients (205 eyes) who had implantation of a MemoryLens U940A IOL from June 1997 to June 2000. The patients were examined using a slitlamp to detect the presence of IOL opacification. In 4 cases, the lenses were explanted because of significant opacification and patient-reported problems; 2 lenses were provided for further analysis. One unused reference MemoryLens U940A IOL was also evaluated. All IOL were stained with von Kossa to determine the presence of calcium in the opacification. To confirm the components presence of an ultraviolet (UV) absorber, the IOLs were examined with an Avatar 330 Fourier transfer infrared (IR) spectroscope and a UV visible spectrophotometer (Philips). The IR spectrums for the IOL were identified using an IR spectrum atlas. The opacified IOLs, reference IOL, and the IOL packaging were further examined to determine the presence of silicone. RESULTS: Various amounts of opacification were found on the MemoryLens U940A IOL in 30 eyes (30 patients) (14.63%). Two explanted IOLs were positive for von Kossa staining, proving the presence of calcium deposits; the reference lens staining was negative. Spectrophotometry showed that the reference IOL and opacified IOLs were of the same polymer. The presence of the UV absorber on the benzophenone base was seen in the reference lens but not the opacified IOLs. In contrast, an increased concentration of low-molecular-weight components generated during the degradation of the polymer was present in the opacified lenses. The white cover pf the IOL is of polydimethyl siloxane, a silicone rubber. However, no silicone rubber was present in any examined lens, perhaps because the IOLs were in contact with alcohol during the histopathologic examination. CONCLUSIONS: The results indicate opacification of the hydrophilic MemoryLens U940A was caused by premature consumption of the UV absorber in the polymer component of the IOLs optic, with a subsequent degradation of the polymer. Whether silicone from the white cover led to the IOL opacification, as reported with other types of hydrophilic IOLs, could not be confirmed. PMID- 15342060 TI - Astigmatism after small-incision clear corneal cataract extraction and intraocular lens implantation in children. AB - PURPOSE: To investigate the magnitude of postoperative astigmatism in children having cataract extraction with intraocular lens (IOL) implantation through a 3.0 mm superior clear corneal incision. SETTING: Department of Ophthalmology, Indiana University School of Medicine, Indianapolis, Indiana, USA. METHODS: This retrospective chart review comprised all pediatric patients having cataract surgery with IOL implantation through a 3.0 mm clear corneal incision from 1997 to 2002. One hundred two eyes of 75 patients were included. All refractions were performed manually by an experienced pediatric ophthalmologist. RESULTS: The mean postoperative retinoscopic cylinder in all patients was 0.63 diopter (D) (range 0.0 to 4.50 D) at 1 month, 0.40 D (range 0.0 to 1.75 D) at 6 months, and 0.51 D (range 0.0 to 2.50 D) at 1 year. Patients aged 0 to 36 months at the time of surgery had a mean postoperative cylinder of 0.22 D at 1 month, 0.03 D at 6 months, and 0.21 D at 1 year. Patients between 36 months and 6 years of age at surgery had a mean refractive cylinder of 0.50 D, 0.38 D, and 0.75 D, respectively. Patients older than 6 years at surgery had a mean refractive cylinder of 0.94 D, 0.75 D, and 0.76 D, respectively. CONCLUSIONS: Small-incision clear corneal cataract extraction with IOL implantation in children led to minimal postoperative astigmatism that remained stable over time. Less astigmatism was observed in children having surgery before they were 36 months old. PMID- 15342061 TI - Prevention of endophthalmitis. AB - PURPOSE: To report the incidence of postoperative endophthalmitis in a series of patients who had cataract surgery with intraocular lens (IOL) implantation and to apply guidelines toward the prevention of postoperative endophthalmitis. SETTING: The Buzard Eye Institute for Corneal, Refractive and Cataract Surgery, Las Vegas, Nevada, USA. METHODS: In this prospective institutional study, 5,131 cataract surgery cases with IOL implantation were performed from 1998 to 2002 by 1 surgeon at a single institute. The surgeon used a blue-line incision at the superior location, povidone- iodine prophylaxis, and postoperative injection of subconjunctival antibiotics. The incidence of endophthalmitis in the study was compared with the general incidence in the United States (range 0.07% to 0.13%) and in published studies (range 0.02% to 0.57%). RESULTS: The 5,131 cases were followed for a mean of 2 years (range 3 months to 4 years). The mean patient age was 69.6 years +/- 10.9 (SD). No case of endophthalmitis occurred. The zero incidence of endophthalmitis was below the general incidence in the United States and in published studies. CONCLUSIONS: The findings suggest that the absence of postoperative endophthalmitis may be related to 4 factors: povidone-iodine prophylaxis, meticulous draping of the eyes, operative technique (blue-line incision), and postoperative injection of subconjunctival antibiotics. PMID- 15342062 TI - Investigation into postoperative endophthalmitis and lessons learned. AB - PURPOSE: To describe an approach to the investigation of a series of endophthalmitis cases to determine whether there was a true outbreak. SETTING: Outpatient facility, Sunderland, England. METHODS: Different approaches to statistical analysis of the probability of infrequent events being due to chance occurrence are described. Potential factors leading to an outbreak were reviewed. RESULTS: Bayesian statistical analysis was shown to be appropriate in the determination of an endophthalmitis outbreak. The only factor found to account for the outbreak was the operating surgeon's recent abandonment of subconjunctival antibiotic prophylaxis. This decision was based on the absence of good evidence that subconjunctival antibiotic injection is effective in prophylaxis. These endophthalmitis cases demonstrate that the absence of evidence for effect is not the same as there being no effect. CONCLUSIONS: Bayesian statistical analysis has a place in determining whether an outbreak has occurred. Withdrawing treatment simply on the basis of a lack of good evidence can lead to undesirable outcomes. PMID- 15342063 TI - Comparative study of first-day postoperative cataract review methods. AB - PURPOSE: To assess whether telephone review on the first day after uneventful phacoemulsification is as effective as postoperative review in a hospital or the patients' home by nursing staff. SETTING: Sunderland Eye Infirmary, Sunderland, United Kingdom. METHOD: This prospective study comprised 3 arms, each of which consisted of 100 patients who had uneventful phacoemulsification with intraocular lens implantation. Patients received the same postoperative medications and were given a questionnaire regarding their opinions on the method of postoperative review. The method of review was different for each arm as follows: (1) home review by a nurse, (2) return to the hospital for review by a nurse, and (3) telephone review by a nurse. Statistical analysis of relative frequencies was done; exact testing was applied throughout to test for differences in proportions and, where applicable, to construct 95% confidence intervals. RESULTS: There were no significant differences between the 3 groups in the degree to which patients understood the instructions and questions. The telephone group was significantly less reassured than the home-visit group, but there was no significant difference in the degree of reassurance between the telephone and hospital-visit groups. Seventy percent of patients in the telephone group listed telephone review as their preferred method of postoperative review. CONCLUSION: In cases of uneventful phacoemulsification, telephone review is safe, effective, and acceptable and is a reasonable alternative to other first-day review methods. PMID- 15342064 TI - Evaluation of biofilm formation on nylon sutures removed from clinically noninfected eyes after cataract surgery. AB - PURPOSE: To determine the presence of bacterial biofilm on nylon sutures removed from clinically noninfected eyes after cataract surgery. SETTING: The Eye Institute, Tan Tock Seng Hospital, Singapore, Singapore. METHODS: Sutures were removed from 10 eyes after cataract surgery at different time periods. Immediately after removal, the sutures were fixed and dehydrated. All sutures were viewed by scanning electron microscopy, and 6 were also viewed by transmission electron microscopy (TEM). RESULTS: There was no evidence of bacterial biofilm formation on the nylon sutures. Significant cellular debris was seen, mainly at the knots. Clusters of coccoid-shaped structures were visible; however, examination by TEM showed they were not bacteria. CONCLUSION: There was no evidence of biofilm formation on sutures removed after cataract surgery from clinically noninfected eyes. PMID- 15342065 TI - Artisan aphakic intraocular lens in children with subluxated crystalline lenses. AB - PURPOSE: To evaluate the results of Artisan (Ophtec) aphakic intraocular lens (IOL) implantation in children with idiopathic subluxated lenses. SETTING: Department of Ophthalmology, Soroka University Medical Center, Beer-Sheva, Israel. METHODS: This retrospective small case series comprised 4 eyes (3 children) with idiopathic essential subluxated lenses that had lens washout, lens capsule removal, Artisan IOL implantation, and peripheral iridectomy. The indications for surgery were reduced visual acuity and monocular diplopia. The main outcome measures were postoperative refraction and change in best corrected visual acuity (BCVA). RESULTS: The postoperative follow-up ranged from 8 to 10 months. After surgery, the BCVA was 6/12 or better in the 3 cases that could be recorded. Visual acuity improved by 2 or more Snellen lines in all 4 eyes. The postoperative spherical equivalent was within +/-1.00 diopter in all cases. No significant postoperative complications were observed. CONCLUSIONS: In 4 eyes with a subluxated crystalline lens, implantation of an Artisan aphakic IOL improved visual acuity. Studies with a larger number of patients and longer follow-up are necessary to confirm these results. PMID- 15342066 TI - Correlation between different measurements within the eye relative to phakic intraocular lens implantation. AB - PURPOSE: To obtain measurements of the white-to-white distance and the anterior chamber and ciliary sulcus diameters in phakic human eyes obtained post-mortem to find a correlation between these measurements. SETTING: David J. Apple, MD Laboratories for Ophthalmic Devices Research, John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: Twenty-two phakic human eyes obtained from eye banks within 24 hours of death were evaluated. The following measurements were performed: white-to-white distance, anterior chamber diameter with a plastic sizer after a small limbal incision, and anterior chamber and ciliary sulcus diameters after fixation and sagittal section of the eyes. The first 10 eyes were studied at the 6 o'clock to 12 o'clock meridian and the last 12 eyes, at the 3 o'clock to 9 o'clock meridian. RESULTS: A positive correlation was found between the white-to-white measurements and the anterior chamber diameter in the 10 eyes studied at the 6 o'clock to 12 o'clock meridian but not in the 12 eyes studied at the 3 o'clock to 9 o'clock meridian. The latter is the meridian frequently used by surgeons to perform white-to-white measurements and thus choose the overall size of the phakic intraocular lens to be implanted. No correlation was found between the white-to-white measurements and the ciliary sulcus diameter in the 2 meridians. CONCLUSION: Experimental studies using cadaver eyes are helpful in evaluating sizing techniques for phakic IOL implantation as well as in better understanding the anatomical relationships between ocular structures. PMID- 15342067 TI - Role of the endothelial pump in flap adhesion after laser in situ keratomileusis. AB - PURPOSE: To investigate the role of the endothelial pump in the attachment of the corneal flap during the early period after laser in situ keratomileusis (LASIK). SETTING: Department of Ophthalmology, Tokyo Dental College, Tokyo, Japan. METHODS: The study comprised 30 porcine eyes. In all eyes, a corneal flap was made with a microkeratome. Five minutes after the flap was placed over the bed, the adhesion strength was measured with a load-testing machine. To inhibit the endothelial pump, ouabain was injected into the anterior chamber (10 eyes) or the endothelium was mechanically scraped (10 eyes). The control group included 10 eyes. RESULTS: The load needed to lift the flap was significantly lower in the ouabain group (0.21 gram force [gf]; P<.01) and the endothelial scrape group (0.22 gf; P<.01) than in the control group (0.61 gf). CONCLUSION: The endothelial pump plays an important role in flap attachment in the early postoperative period after LASIK. PMID- 15342068 TI - Spring constants of capsular tension rings. AB - PURPOSE: To determine the spring constants of several types of capsular tension rings (CTRs). SETTING: Department of Ophthalmology, Johannes Gutenberg University, Mainz, Germany. METHODS: Fourteen models of CTRs provided by 9 companies were tested; all were 11.0 mm in diameter. The CTRs were positioned in a special retainer of a material-testing machine and compressed at a constant speed of 10.0 mm per minute. The distance and force values were recorded, and each CTR model was tested 3 times using a new specimen each time. The resulting measurements were used to calculate a mean spring constant for each CTR model. RESULTS: All tested CTRs showed linear proportionality between force and compression distance. Test results were repeatable and precise. Capsular tension rings of the same model had comparable mechanical properties, and different models had different mean spring constants ranging from 0.8162 to 4.5501 mN/mm. CONCLUSIONS: The spring constant of a CTR is a suitable mechanical characteristic to facilitate the choice of CTR model. A CTR with a low spring constant may be advantageous for the management of zonular dialysis, whereas a CTR with a higher spring constant may reduce capsular bag shrinkage more effectively. PMID- 15342070 TI - Beyond the P. I: Problems with probability. PMID- 15342069 TI - Decreased keratocyte death after laser-assisted subepithelial keratectomy and photorefractive keratectomy in rabbits. AB - PURPOSE: To compare keratocyte loss in the corneal stroma after laser-assisted subepithelial keratectomy (LASEK) and photorefractive keratectomy (PRK) in rabbits. SETTING: Department of Ophthalmology, University of Essen, Essen, and the Institute of Anatomy, University of Bochum, Bochum, Germany. METHODS: Laser assisted subepithelial keratectomy and PRK were performed in rabbits and studied 1, 3, 10, and 20 days after surgery. Excimer photoablation was done unilaterally with a 6.0 mm ablation zone and an 80 microm depth, equivalent to -6.0 diopters. Keratocyte death was analyzed using DNA fragmentation-detecting terminal deoxynucleotidyl transferase deoxy-UTR-nick end labeling (TUNEL) assay and transmission electron microscopy. RESULTS: Numerous TUNEL-positive keratocytes occurred 1 day after PRK; the number decreased significantly after 3 days. After LASEK, significantly fewer TUNEL-positive keratocytes were noted at the early time points (P<.001 at 1 day; P< or =.05 at 3 days). At 10 days, the number of TUNEL-positive keratocytes decreased in both groups but remained significantly higher after PRK than after LASEK (P<.001). Twenty days after both procedures, no significant signs of keratocyte death were found in the corneal stroma. Transmission electron microscopy revealed few apoptotic keratocytes after LASEK. After PRK, apoptotic keratocytes, characterized by chromatin condensation, apoptotic bodies, and cell shrinkage, were scattered in the stroma. The ultrastructural findings confirmed the results obtained with the TUNEL assay. CONCLUSIONS: Laser-assisted subepithelial keratectomy induced significantly less apoptotic keratocyte death than PRK and promoted wound healing in the acute phase after photoablation. This procedure may offer the possibility of treating higher myopia with a decreased risk for developing wound healing-related complications known to occur after PRK. PMID- 15342071 TI - Contact between 3 phakic intraocular lens models and the crystalline lens: an anterior chamber optical coherence tomography study. AB - Three phakic intraocular lens (IOL) models were implanted in 3 different patients. With the usual slitlamp examination, it was not possible to determine whether there was contact between the IOLs and the natural crystalline lens. Using the anterior chamber optical coherence tomography (AC OCT) scanner, direct contact between the natural crystalline lens and the 3 phakic IOLs was revealed. A dynamic study of the contact was performed during accommodation. These observations show that examination of the anterior segment with the AC OCT scanner provides new data about the status of the anterior segment after implantation of phakic IOLs. PMID- 15342072 TI - Spontaneous dislocation of a phakic refractive lens into the vitreous cavity. AB - A 36-year-old woman with high myopia had uneventful implantation of a phakic refractive lens (PRL) bilaterally. Two months postoperatively, the best corrected visual acuity (BCVA) in the right eye decreased to the preoperative level and the posterior chamber PRL disappeared from the anterior segment and was found lying in the vitreous cavity inferiorly. After lensectomy and pars plana vitrectomy, the PRL was removed through the initial clear corneal incision, improving the BCVA to 1.0. A zonular defect associated with high myopia, previously forgotten and unrecognized ocular trauma, or intraoperative manipulations may have resulted in the spontaneous dislocation of the PRL. PMID- 15342073 TI - Bimanual bipolar diathermy for recurrent hyphema after anterior segment intraocular surgery. AB - Phacoemulsification with mechanical pupil dilation was performed on a functionally monocular glaucoma patient with pseudoexfoliation syndrome. The postoperative course was complicated by persistent intraocular hemorrhaging from the pupil margin in multiple locations that ceased temporarily with a marked elevation in intraocular pressure (IOP). Normalization of IOP with medication or paracentesis resulted in recurrent bleeding and a subsequent increase in IOP elevation. Surgical intervention using bipolar diathermy was required to control the bleeding and the elevated IOP. A bimanual approach allowed the corrective procedure to be performed in a simple and efficacious manner. PMID- 15342074 TI - Particles at the laser in situ keratomileusis flap interface. PMID- 15342075 TI - Perioperative prophylaxis for cataract surgery: survey of ophthalmologists in the north of England. PMID- 15342076 TI - Workshop on trichothecenes with a focus on DON: summary report. AB - A number of mycotoxins of the class of trichothecenes are produced by a variety of Fusarium fungi commonly found on cereals. Unfavourable weather conditions may lead to a high level of Fusarium infections in crops such as wheat and correspondingly high trichothecene contents. The ILSI Europe Natural Toxin Task Force therefore organised a workshop on trichothecenes with a special focus on deoxynivalenol (DON). A number of experts reviewed the current knowledge on trichothecenes with respect to occurrence, including aspects of mould growth, toxin formation, storage and effects of processing; prevention; analytical methodologies, including sampling; surveillance and exposure assessments; and toxicology and risk assessment. A number of recommendations were given under the headings: prevention, sampling and analytical methods, exposure assessment, and toxicology. Gaps in knowledge were also identified. PMID- 15342077 TI - Fusarium taxonomy with relation to trichothecene formation. PMID- 15342078 TI - Influence of agricultural practices on fusarium infection of cereals and subsequent contamination of grain by trichothecene mycotoxins. AB - Fusarium head blight (FHB) of small grain cereals and ear rot in maize are significant diseases across the world. Infection can not only result in reduced yield as a result of shrunken grains but also result in reduced milling and malting quality and the contamination of grains with mycotoxins. Mycotoxins are hazardous to animal and human health. Therefore, guidelines and legislation are in place, or under consideration, in most countries to protect consumers and animal welfare. As fusarium mycotoxins are produced within the growing crop, it is important to understand how agricultural practices affect mycotoxin contamination of grain. Such information could then be used to determine guidelines on "Good Agricultural Practice" (GAP) to minimise the mycotoxin contamination of cereal products. Evidence is provided to show the importance of choice of cultivar, crop rotation, soil cultivation, fertiliser and the chemical and biological control of insects, weeds and fungi. PMID- 15342079 TI - Resistance in wheat to Fusarium infection and trichothecene formation. AB - The state of the art in Fusarium head blight resistance research is reviewed with reference to breeding for genetic resistance to Fusarium in wheat in practice. Fusarium graminearum and F. culmorum produce the trichothecene mycotoxin deoxynivalenol (DON). DON has phytotoxic properties and is an important aggressiveness factor in head blight. Head blight resistance in wheat is not specific for either F. graminearum or F. culmorum. Resistance components include resistance to penetration, resistance to colonization and mechanisms that influence kernel DON content. The resistance to Fusarium in wheat is a quantitative trait with relative high heritability and controlled by a few genes with major effect. A major quantitative trait locus (QTL) for head blight resistance from the Chinese variety Sumai 3 has been identified and verified by several research groups via molecular marker analysis. Research is now directed at identifying additional QTLs to make accumulation of resistance genes in elite wheat lines possible. The policy of official variety list trials may affect the head blight resistant level of future wheat varieties by excluding candidate varieties that are a too susceptible to Fusarium. A higher level of Fusarium head blight will guarantee lower risks for the farmer of crop loss due to reduced grain yield, low quality and mycotoxin contamination. PMID- 15342080 TI - Influence of harvest and storage conditions on trichothecenes levels in various cereals. AB - Grain should be harvested at lowest possible humidity, or immediately dried to safe storage humidity to prevent the growth of fungi. Critical steps of grain harvest like conditions of used combine harvester, temperatures and aeration are discussed on their impact on mycotoxin formation. PMID- 15342081 TI - Influence of processing on trichothecene levels. AB - Trichothecene mycotoxins frequently occur in cereal grains that are intended for food production. The process of converting grains into food and drinks for consumers has significant effects on the levels of toxins in the final food. Surveillance of retail food and drinks of cereal origin demonstrates that trichothecenes do survive the production processes employed. Trichothecenes are relatively heat stable chemicals, with high water solubility, properties that affect their processing fate. It is known that the extent of transmission into final food products is dependent on the pattern of Fusarium infection in the grains. For dry-milled products, the most highly contaminated fractions are those that contain the whole or the outer portions of the grain. For wet milling, the trichothecenes primarily transfer to the aqueous fractions, the most contaminated streams enter the animal food chain, although there is no evidence of significant trichothecenes transmission into animal products. However, anomalies remain in the understanding of the processing effects of several major cereal processes used in European food production (e.g. baking and brewing). PMID- 15342082 TI - Cellular and molecular mechanisms for immune modulation by deoxynivalenol and other trichothecenes: unraveling a paradox. AB - Macrophages, T cells, and B cells of the immune system are central targets of deoxynivalenol (DON) and other trichothecenes-mycotoxins that can be immunostimulatory or immunosuppressive depending on dose, exposure frequency and timing of functional immune assay. Notably, low dose trichothecene exposure transcriptionally and post-transcriptionally upregulates expression of cytokines, chemokines and inflammatory genes with concurrent immune stimulation, whereas high dose exposure promotes leukocyte apoptosis with concomitant immune suppression. DON and other trichothecenes, via a mechanism known as the ribotoxic stress response, bind to ribosomes and rapidly activate mitogen-activated protein kinases (MAPKs). The latter are important transducers of downstream signaling events related to immune response and apoptosis. Using cloned macrophages, our laboratory has identified two critical upstream transducers of DON-induced MAPK activation. One transducer is double-stranded RNA-(dsRNA)-activated protein kinase (PKR), a widely-expressed serine/theonine protein kinase that can be activated by dsRNA, interferon, and other agents. The second transducer is hematopoetic cell kinase (Hck), a non-receptor associated Src family kinase. Inhibitors and gene silencing studies have revealed that Hck and PKR play roles in DON induced gene expression and apoptosis. Future studies should focus on the molecular linkages between these kinases and trichothecene toxicity. PMID- 15342083 TI - Haematotoxicity of trichothecenes. AB - Human toxicosis induced by consumption of foodstuffs contaminated with trichothecenes presents one common major symptom: a haematological perturbation manifesting principally as thrombocytopenia and leukopenia. The patients have rapidly progressing coagulation problems, and compromised resistance to infections. Consequently, they are subject to septicaemia and massive haemorrhages. In horses, cattle, poultry, cats, mice and guinea pigs, subacute and subchronic ingestion of trichothecenes causes a decrease of circulating blood cells frequently associated with bone marrow failure. The origins of haematological effects observed in Fusarium toxin intoxications have been elucidated using in vitro tests. Haematopoietic progenitors are the main target of trichothecenes. Deoxynivalenol (DON) is the least myelotoxic and T-2 the most. As circulating blood cells present a less important sensitivity to these toxins, haematological troubles observed in toxicosis are due to myelotoxicity of these toxins. PMID- 15342084 TI - Toxicity data relevant for hazard characterization. AB - The present paper summarizes toxicity data relevant for hazard characterization for the trichothecene mycotoxins deoxynivalenol (DON), nivalenol (NIV), T-2 and HT-2 from recent opinions prepared by the European Commission Scientific Committee on Food (SCF) and the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Details on immunotoxicity, cardiovascular toxicity and co-occurrence of different trichothecenes and other mycotoxins and their possible interactions are considered in separate papers in the present issue as well as other aspects such as mould growth, trichothecenes formation, storage, processing, sampling, analytical measurements, exposure assessment and surveillance. The toxicological profiles of DON, NIV, T-2 and HT-2 are similar. The general toxicity and immunotoxicity in experimental animals, and for NIV also haematotoxicity, are considered to be the critical effects. Tolerable Daily Intakes of 1, 0.7 and 0.06 microg/kg b.w. were established for DON, NIV and the sum of T-2 and HT-2, respectively. The TDI's for NIV, T-2 and HT-2 were made temporary because of deficiencies the database. PMID- 15342085 TI - Combined toxic effects of mycotoxins. AB - It is known for many years that several food items, derived from plants infected by fungi in the field during growing of the plant or during harvest and storage of the food item, can contain concomitantly different mycotoxins. As these combined mycotoxins occur simultaneously in the food item, consumption of the food will lead to a combined intake depending on the absorption rates of the different mycotoxins. Therefore, the question is justified whether such a combined intake of mycotoxins would lead to a possible higher risk for adverse health effects than the intake of one of these mycotoxins alone. It will be dealt with on the basis of some practical cases of such combined intake of mycotoxins of which research data are available. This is the case for citrinin and ochratoxin A, but as the workshop focuses on trichotecenes and so this paper concentrates on these. When the mycotoxins are of similar structure and of the same species, or of the same families, it is likely to expect that the mode of action of the mycotoxins and or the toxicity profiles will be quite similar. This indicates that such related mycotoxins are likely to exert only additive effects, which is important to know. In terms of risk assessment, these mycotoxins could be dealt with by establishing a group daily tolerable intake (TDI) or a provisional tolerable weekly intake (PTWI). In terms of risk assessment those mycotoxins which interact in synergistic manner are of more concern. It is concluded that, at present tools are not fully developed to establish the type of interaction or whether there is any interaction at all. PMID- 15342086 TI - Novel sampling methods for the analysis of mycotoxins and the combination with spectroscopic methods for the rapid evaluation of deoxynivalenol contamination. AB - A novel non-destructive sampling approach is described for the identification of food matrices contaminated with deoxynivalenol and other mycotoxins. This technique is different from currently applied sampling procedures for this purpose and is based on the principle that surface material from the tested goods is collected on a filter and brought to chemical or spectroscopic analysis. This approach has been applied to several matrices and mycotoxins, with a focus on those mycotoxin-matrix combinations that are of main relevance due to current or future legislation. Tests were carried out with a facility that has been shown to be suitable to process large quantities of materials at points of transaction, such as harbour. Further experiments with a small sampling lance prototype showed that analytical results from the chemical analysis of the tested goods can be correlated with the results obtained with this novel sampling procedure. PMID- 15342087 TI - State of the art of trichothecenes analysis. AB - Methods to analyze trichothecenes should be fast, reliable and economical. The known methods can be divided into two categories: the 'instrumental methods' as gas chromatography (GC) and high performance liquid chromatography (HPLC) and the so called 'fast methods' like thin layer chromatography (TLC), enzyme-linked immunosorbant analyses (ELISA) and flow through immunoassays. The most frequently used instrumental methods for the determination of deoxynivalenol (DON), HT-2- and T2-toxin in cereals and cereal products are based on gas chromatography with electron capture detectors or mass spectrometric (MS) detectors. More than 70% of the literature published in the last decade focuses on these techniques. Recently, high performance liquid chromatography with fluorescence detection and with mass spectrometric detection have gained importance. Up to now, a method which is able to detect types A and B trichothecenes simultaneously with a sufficiently low limit of detection is lacking, even though methods are published which have at least the same clean up step. PMID- 15342088 TI - Rapid methods for deoxynivalenol and other trichothecenes. AB - Method development for deoxynivalenol (DON) and other trichothecenes in recent years was driven by the analytical necessities arising from its widespread (and increasing) occurrence in foods and feeds. This has resulted in the establishment of guideline levels for animal feed, tolerable daily intake (TDI) levels for humans, and most importantly, in the prospect of low-tolerance levels for these toxins in foods in the near future. In order to ensure reliable determination of the toxin content at the tolerance levels, routine analytical methods must have detection limits of less than the tolerance level. This paper intends to give an overview of current analytical developments of rapid testing for deoxynivalenol and other trichothecene mycotoxins, with a special focus on antibody-based techniques. This includes high-throughput instrumental analysis for the laboratory environment, as well as rapid visual tests for on-site testing. The applicability of rapid tests within an integrated detection system for mycotoxins in foods is discussed. PMID- 15342089 TI - Trichothecenes: reference materials and method validation. AB - The frequent contamination of food and feed with trichothecene mycotoxins, the high consumption of these products, and the potential risk associated herewith, has led to an increasing public awareness and therefore to the establishment of measures to control trichothecene contamination. The analytical difficulty and the economic importance of controlling trichothecenes in food and feed support the need for certified reference materials (CRMs) and validated methods. They form invaluable tools to ensure comparability and traceability in analytical measurements and are very useful for the implementation of written standards, legislation/regulations and laboratory accreditation. The present paper provides an overview of previous work, current strategies and prospectives for the production of CRMs and validation of analytical methods in the field of trichothecene analysis. Additional information is given on methodological demands, normative frameworks and commonly accepted procedures. PMID- 15342090 TI - Report from SCOOP task 3.2.10 "collection of occurrence data of Fusarium toxins in food and assessment of dietary intake by the population of EU member states". Subtask: trichothecenes. AB - In 2001 the SCOOP (SCOOP: Scientific Co-operation on Questions relating to Food) task 3.2.10 "Collection of occurrence data of Fusarium toxins in food and assessment of dietary intake by the population of EU Member States" was established. The task was divided in three subtasks (zearalenone, fumonisins and trichothecenes). Results of the subtask trichothecenes, which is co-ordinated by The Netherlands, will be presented. About 35,000 results were received about occurrence of 12 different trichothecenes (deoxynivalenol (DON), nivalenol (NIV), 3 and 15 acetyl-deoxynivalenol (3/15-AcDON), fusarenon-X (FUS-X), T-2 and HT-2 toxin, T2-triol, diacetoxyscirpenol (DAS), neosolaniol (NEOSOL, monoacetoxyscirpenol (MAS) and verrucarol (VOL)) in various food and food raw materials from 12 countries. Only the results of DON, NIV, T-2 and HT-2 toxin are included in this paper, because most of the data refer to these toxins and only for these trichothecenes the Scientific Committee for Food sets (temporary) Tolerable Daily Intakes (t-TDIs). Occurrence data: By far most of the occurrence data were obtained for DON in wheat. Among cereals, corn showed the highest level of contamination with trichothecenes. Consumption data: There is a significant lack of consumption data in some countries. In particular information on baby's and children's food is generally not available. Intake data: Wheat and wheat containing products (like bread and pasta) represent the major source of intake for the four trichothecenes. The mean intakes for DON are below the TDI, however for the young children groups the mean intakes are sometimes (very) close to the TDI. By comparing the high intake levels for DON with the TDI, it is clear that especially for the young children groups most of the intakes are above the TDI. For NIV, the (mean and high level) intakes are far below the TDI. The summarised T-2 and HT-2 toxin intakes are in most cases (for the mean as well as the high level intake) above the t-TDI. PMID- 15342091 TI - Reduced intake of deoxynivalenol in The Netherlands: a risk assessment update. AB - In 1998 and 1999, wheat contained high levels of deoxynivalenol (DON) in The Netherlands. Eighty percent of the 1-year-old children exceeded the TDI (20% exceeded 2TDI). Assuming a long-term intake a negative effect on body weight might be possible. In 1999 the Dutch government took several risk management measures. We performed a probabilistic exposure and effect assessment and compared current and previous DON exposure and potential health effects for young children. The DON contamination of wheat was reduced for 50%. The estimated DON intake was reduced to one-third of the previous levels. One-year-old children showed the highest DON-intake with a median of 0.46 microg/kgbw/day (95th percentile: 1.00 microg/kgbw/day). The probabilistic effect assessment showed that at the 95th percentile of DON-intake a reduced body weight was unlikely to exceed 9%. Taking into account the possible (partly) reversibility of this effect we conclude that no clear adverse health effects will be associated to the exposure (February 2000-December 2002) to DON in The Netherlands. PMID- 15342092 TI - The risk assessment paradigm and its application for trichothecenes. AB - Risk analysis for trichothecene mycotoxins and other food contaminants, which are to a significant extent unavoidable, presents considerable challenges. Risk assessment is constrained by uncertainties associated with the lack of adequate data, and risk management must consider the fact that mycotoxin contamination can have serious impacts on trade and food sufficiency. These factors necessitate good communication between the risk assessors and risk managers in formulating the questions to be addressed by the risk assessment. Risk assessment must be an iterative process, since the problem formulation and the risk assessment may need to be revised to reflect new data and theories. In addition to providing advice to risk managers, risk assessment should provide a blueprint for future research by illustrating what observations will influence a prediction. The international risk assessments completed for deoxynivalenol, T-2 and HT-2 toxins, and nivalenol have noted a number of issues regarding the lack of adequate intake data for exposure assessment and significant gaps in toxicological studies for hazard characterizations. Addressing these uncertainties would provide risk managers with better guidance for control measures. PMID- 15342093 TI - Prevention strategies for trichothecenes. AB - Contamination of cereal commodities with mycotoxins represents a significant hazard to consumer health and has thus received increasing attention from food safety authorities and legislators. For trichothecenes, and deoxynivalenol (DON) in particular, the imminent implementation of legislative limits has focused attention on ways to prevent entry of such mycotoxin contaminants into the food and feed chains. Knowledge of the pre- and post-harvest stages in the cereal production chain and in particular information on where prevention strategies can be implemented is being used to develop quality assurance systems for improving food safety. Information on the ecology of Fusarium species, breeding for resistance, more effective fungicides, potential for biological control and effective drying and storage and preservation systems, are all helping to develop effective preventative strategies for minimising consumer exposure to trichothecenes and other mycotoxins. PMID- 15342094 TI - Deoxynivalenol in cereals in Russia. AB - A survey of the occurrence of deoxynivalenol (DON) and zearalenone (ZEN) in wheat, rye, barley and maize harvested in 1989-2001 in several regions of Russia has been conducted. A total of 5652 samples of cereals were analysed for DON and ZEN by using TLC and normal-phase HPLC with UV-detector. DON was detected in 69% of 2166 samples from Krasnodar region which is considered to be the major Fusarium endemic region of Russia. The contamination levels ranged from 0.1 till 8.6 ppm, MTEL was exceeded in 37% of these samples. The positive correlation between DON concentration and a percentage of Fusaria-damaged wheat kernels has been shown. DON occurrence and contamination levels were much lower that for wheat. Based on the results of monitoring and the data of average actual consumption of wheat products in Russia, the estimated daily intake of DON per 1 kg of body weight (EDI)was calculated. EDI varied from 0.07 ug in 1990-1991 till 1.40 ug in 1992. Although average EDI were lower than adopted tolerable daily intake (TDI, 3 ug/kg body weight) EDIs for the North-Caucasian region in some cases exceeded TDI. PMID- 15342095 TI - Reduction of deoxynivalenol during durum wheat processing and spaghetti cooking. AB - The reduction of deoxynivalenol (DON) during durum wheat processing and spaghetti cooking was investigated in nine samples of durum wheat contaminated with DON under field conditions (either naturally contaminated or artificially inoculated with Fusarium) at levels ranging from 0.3 to 13.1 microg/g. A consistent reduction of DON levels was observed during each of the processing steps from uncleaned durum wheat to cooked spaghetti. With respect to the uncleaned wheat, the average levels of DON were 77% in cleaned wheat, 37% in semolina, 33% in spaghetti and 20% in cooked spaghetti, with relative standard deviations of 11%, 13%, 12% and 8%, respectively. Average DON levels in the screenings, bran and fine middlings were 4.1-, 1.6- and 0.6-fold, respectively, relative to the uncleaned wheat. DON was almost completely recovered from spaghetti after cooking, with a repartition between cooked spaghetti (without the corresponding DON amount from absorbed water) and cooking water generally in favour of the latter. An increased DON leaching into the cooking water was observed when a higher water to spaghetti ratio was used during cooking. This study strongly supports the basis of previous knowledge on DON distribution in durum wheat, semolina and pasta products, and concludes that the retention level of DON from grains on the market to cooked pasta in the plate can be conservatively assessed at 25% or less. PMID- 15342096 TI - Layer-specific potentiation of evoked IPSCs in rat hippocampal CA1 pyramidal cells by lanthanum. AB - Lanthanum (La3+) potentiates or depresses GABAA receptors (GABAAR) in a subunit dependent manner. Such differential modulators may help to discriminate between layer-specific inhibitory synaptic inputs to individual neurons, which use different molecular GABAAR isoforms. Here, we show that inhibitory postsynaptic currents (IPSCs) in CA1 pyramidal cells are potentiated by La3+ (100 microM) when they are evoked by stimulation in stratum oriens. In contrast, stimulation in stratum radiatum yields IPSCs which are not sensitive towards La3+. These data point towards an input-specific molecular and functional diversity of inhibitory synapses at CA1 pyramidal cells. PMID- 15342097 TI - A [17F]-fluoromethane PET/TMS study of effective connectivity. AB - We used transcranial magnetic stimulation (TMS) in combination with positron emission tomography (PET) to investigate the effective connectivity of four cortical regions within the same study. By employing [17F]-[CH3F] ([17F] fluoromethane) as a radiotracer of blood-flow, we were able to obtain increased sensitivity compared to [15O]-H2O for both cortical and subcortical structures. The brain areas investigated were left primary motor cortex, right primary visual cortex, and left and right prefrontal areas. We found that each site of stimulation yielded a different pattern of activation/deactivation consistent with its anatomical connectivity. Moreover, we found that TMS of prefrontal and motor cortical areas gave rise to trans-synaptic activation of subcortical circuits. PMID- 15342098 TI - Neural substrates of the performance of an auditory verbal memory: between subjects analysis by fMRI. AB - We studied the neural correlates for the performance of auditory verbal memory using fMRI in 28 normal right-handed volunteers who had to encode and retrieve 50 related word-pair associates. Since the effectiveness of encoding and retrieval processes contribute to memory performance, we combined them in one analysis in addition to examining each condition. Between-subjects analyses were performed by a two-sample t-test in which brain activation in the good-performance group was compared with that of the poor-performance group. Regression analysis was also carried out to find the regions whose activations were linearly correlated to the number of correct recalls by all subjects. In addition to the positive correlates of memory performance, the areas whose activations have a negative effect were also examined. The results indicate that the brain networks of positive correlates including the parahippocampal gyrus and negative correlates including the right prefrontal cortex constitute neural substrates for the performance of auditory verbal memory. PMID- 15342099 TI - Ameroid rings for gradual chronic constriction of the sciatic nerve in rats: contribution of different nerves to neuropathic pain. AB - Mononeuropathy was induced by placing an ameroid ring around the sciatic nerve and was compared with chronic constriction injury (CCI) of the sciatic nerve [Pain 33 (1988) 87] in rats. Mechanical allodynia was assessed and the role of sciatic and saphenous afferents (Adelta and C) in thermal hyperalgesia investigated. A shorter duration of mechanical allodynia in ameroid rats as compared to CCI rats was observed. Thermal hyperalgesia was observed in the saphenous innervated skin of the hindpaw for Adelta and C nociceptors in ameroid and for Adelta nociceptors only in CCI rats, respectively. The sciatic innervated skin showed a thermal hypoalgesia with a fast onset for Adelta afferents and a slower onset for C afferents in CCI and ameroid rats. The duration of both thermal hypo- and hyperalgesia was longer in ameroid rats. We conclude that ameroid rings are a useful tool for the investigation of long-duration hyperalgesic effects of nerve injury, as the effects were more stable and seen for a longer time (>8 weeks) as compared to the CCI model. The uninjured saphenous afferents, in particular C fibers, mediate thermal hyperalgesia after chronic constriction of the sciatic nerve using an ameroid ring. PMID- 15342100 TI - NR2B receptors are involved in the mediation of spinal segmental reflex potentials but not in the cumulative motoneuronal depolarization in vitro. AB - Windup, the frequency dependent build-up of spinal neuronal responses is an electrophysiological model of the development of the central sensitization in the chronic pain states. NR2B subunit containing NMDA-type glutamate receptors are implicated in the windup of dorsal horn neurons, while their role at the motoneuronal level is controversial. The cumulative motoneuronal depolarization in hemisected rat spinal cord preparation is an in vitro model of windup. The role of NR2B receptors in this process, and in the mediation of dorsal root stimulation evoked ventral root reflex potentials was elucidated. Three selective NR2B antagonists; CP-101,606; CI-1041 and Co-101244 (1 microM) were used. They had only weak, but statistically significant inhibitory effect on the early part of ventral root response, and did not influence the cumulative depolarization. On the contrary, non-selective NMDA antagonist APV (40 microM) decreased both responses markedly. We conclude that the pharmacological sensitivities of windup at the sensory and motor levels are different. NR2B containing NMDA receptors have major role in the mediation of the windup of dorsal horn neurons, but their contribution to this phenomenon at the motor level is negligible. PMID- 15342101 TI - Beta-hydroxy-butyrate alters the extracellular content of S100B in astrocyte cultures. AB - Astrocytes have a variety of roles in maintaining neural tissue physiology, including energetic support, uptake and metabolism of glutamate and secretion of neurotrophic factors. Glutamate toxicity has been implicated in neurodegenerative disorders associated with conditions related to energy failure, and to elevation of glutamate extracellular levels in brain. Glucose is the main energetic substrate for brain cells but, in some circumstances, the ketone bodies are used as a supplementary source and have been suggested to be neuroprotective agents against seizure disorders. Here, we investigate some possible biochemical changes in astrocyte cultures induced by beta-hydroxy-butyrate, the predominant blood ketone body. Its effect upon S100B secretion, astrocyte morphology and glutamate uptake was particularly investigated. S100B, a calcium-binding protein expressed and secreted by astrocytes, has neurotrophic activity and a possible role in epileptogenesis. Cell morphology was investigated by phase-contrast microscopy and immunocytochemistry for actin, GFAP and S100B. Our data show that beta hydroxy-butyrate induces dramatic changes in astrocyte morphology and, independent of this, causes changes in the extracellular content of S100B. We observed an increment in S100B 1 h after beta-hydroxy-butyrate addition and a decrease 24 h later. No changes were observed in glutamate uptake. These astrocytic modifications may be associated with reduced neuronal excitability observed in the ketogenic condition. PMID- 15342102 TI - Ultrastructural and quantitative age-related changes in capillaries of the dorsal lateral geniculate nucleus. AB - An ultrastructural and quantitative study of age-related changes in the capillaries of the dorsal lateral geniculate nucleus was carried out using male Wistar rats aged 3, 24, and 28 months. The most important structural changes were found in the basal lamina: thickenings either homogeneously distributed or in specific points; spurs towards the astrocyte sheath; and osmiophilic membrane like inclusions located within the basal lamina. Endothelial cells and pericytes showed an increase in inclusions and dense bodies in the cytoplasm. The quantitative study showed that the most pronounced alteration was the thickening of the basal lamina, which existed at 24 months. Later, at 28 months, thinning of the endothelial cells was observed together with an increase in mitochondria size and the number of pinocytic vesicles. These changes could be an endothelial cell response to compensate for the increasing transport difficulties caused by the thickening of the basal lamina. The progressive age-related changes observed in the structure of the capillaries might have an effect on the regulation of blood and brain tissue exchanges, and thus might contribute to the development of degenerative alterations in surrounding aging neurones. PMID- 15342103 TI - Neuroprotective effect of L-DOPA co-administered with the adenosine A2A receptor agonist CGS 21680 in an animal model of Parkinson's disease. AB - Adenosine A2A receptors are a new target for drug development in Parkinson's disease. Some experimental and clinical data suggest that A2A receptor antagonists can provide symptomatic improvement by potentiating the effects of L DOPA as well as a decrease in secondary effects such as L-DOPA-induced dyskinesia. L-DOPA-induced behavioral sensitization in unilateral 6 hydroxydopamine-lesioned rats is frequently used as an experimental model of L DOPA-induced dyskinesia. In the present work this model was used to evaluate the effect of the A2A receptor agonist CGS 21680 and the A2A receptor antagonist MSX 3 on L-DOPA-induced behavioral sensitization and 6-hydroxydopamine-induced striatal dopamine denervation. L-DOPA-induced behavioral sensitization was determined as an increase in L-DOPA-induced abnormal involuntary movements and enhancement of apomorphine-induced turning behavior. Striatal dopamine innervation was determined by measuring tyrosine-hydroxylase immunoreactivity. Chronic administration of MSX-3 was not found to be effective at counteracting L DOPA-induced behavioral sensitization. On the other hand, CGS 21680 completely avoided the development of L-DOPA-induced behavioral sensitization. The analysis of the striatal dopamine innervation showed that L-DOPA-CGS 21680 co-treatment conferred neuroprotection to the toxic effects of 6-hydroxydopamine. This neuroprotective effect was dependent on A2A and D2 receptor stimulation, since it was counteracted by MSX-3 and by the D2 receptor antagonist haloperidol. These results open new therapeutic avenues in early events in Parkinson's disease. PMID- 15342104 TI - Galantamine and nicotine have a synergistic effect on inhibition of microglial activation induced by HIV-1 gp120. AB - Chronic brain inflammation is the common final pathway in the majority of neurodegenerative diseases and central to this phenomenon is the immunological activation of brain mononuclear phagocyte cells, called microglia. This inflammatory mechanism is a central component of HIV-associated dementia (HAD). In the healthy state, there are endogenous signals from neurons and astrocytes, which limit excessive central nervous system (CNS) inflammation. However, the signals controlling this process have not been fully elucidated. Studies on the peripheral nervous system suggest that a cholinergic anti-inflammatory pathway regulates systemic inflammatory response by way of acetylcholine acting at the alpha7 nicotinic acetylcholine receptor (alpha7nAChR) found on blood-borne macrophages. Recent data from our laboratory indicates that cultured microglial cells also express this same receptor and that microglial anti-inflammatory properties are mediated through it and the p44/42 mitogen-activated protein kinase (MAPK) system. Here we report for the first time the creation of an in vitro model of HAD composed of cultured microglial cells synergistically activated by the addition of IFN-gamma and the HIV-1 coat glycoprotein, gp120. Furthermore, this activation, as measured by TNF-alpha and nitric oxide (NO) release, is synergistically attenuated through the alpha7 nAChR and p44/42 MAPK system by pretreatment with nicotine, and the cholinesterase inhibitor, galantamine. Our findings suggest a novel therapeutic combination to treat or prevent the onset of HAD through this modulation of the microglia inflammatory mechanism. PMID- 15342105 TI - Parabrachial inputs to Fos-immunoreactive neurons in the lateral central nucleus of amygdala activated by hypotension: a light and electron microscopic study in the rat. AB - Morphological features and functional implications of projections of the parabrachial nucleus to the central nucleus of the amygdala were investigated in the rat. The anatomical study was based on injections of the tracers horseradish peroxidase and biotinylated dextran amine. An extremely dense concentration of labeled fibers was found in the lateral and lateral capsular subdivisions of the central nucleus of the amygdala, originating mainly from the external lateral and ventral lateral subnuclei of the parabrachial nucleus. The parabrachial fibers exhibited the morphological characteristic of forming dense pericellular terminal arborizations. The functional implications of this pathway in cardiovascular functions were verified using Fos protein induction in response to hypotension induced by continuous intravenous administration of hydralazine-hydrochloride. In this paradigm, Fos immunoreactivity was found to be confined to the lateral and lateral capsular subdivisions of the central nucleus of the amygdala. Double immunostaining methods were used to visualize, at the electron microscopic level, terminals labeled by biotinylated dextran amine and Fos cell labeling. With this approach, we were able to confirm that Fos-immunoreactive neurons in the central nucleus of the amygdala receive axosomatic terminals from the parabrachial nucleus. The present findings point out that parabrachial inputs to the central nucleus of the amygdala play a relevant role in regulating cardiovascular function. PMID- 15342106 TI - Involvement of 5-HT1A and 5-HT2 receptors of the dorsal periaqueductal gray in the regulation of the defensive behaviors generated by the elevated T-maze. AB - Previous studies have shown that serotonin plays an inhibitory role in escape behavior induced by the aversive stimulation of the dorsal periaqueductal gray matter (DPAG). This defensive behavior has been related to panic disorder. Serotonin injected into the DPAG also inhibits escape behavior generated by the elevated T-maze. Besides escape, this test also measures inhibitory avoidance, a behavior associated with generalized anxiety disorder. We presently evaluate the role of the 5-HT1A, 5-HT2A and 5-HT2C receptors of the DPAG in the modulation of inhibitory avoidance and escape responses of rats submitted to the elevated T maze. The results showed that intra-DPAG administration of the 5-HT1A receptor antagonist WAY-100635 and of the preferential antagonists of 5-HT2A and 5-HT2C receptors, ketanserin and SDZ SER 082, respectively, did not change rat behavior in the elevated T-maze. Intra-DPAG injection of serotonin inhibited escape, an effect blocked by local injection of these three antagonists. Ketanserin and SDZ SER 082, but not WAY-100635 antagonized the effect of serotonin in facilitating inhibitory avoidance. Intra-DPAG injection of the 5-HT1A agonist 8-OH-DPAT and of DOI, a preferential 5-HT2A agonist, also inhibited escape, an effect antagonized by WAY-100635 and ketanserin, respectively. The results indicate that serotonin in the DPAG exerts a phasic regulatory control on inhibitory avoidance and escape behaviors in the elevated T-maze. 5-HT1A and 5-HT2C receptors in the DPAG play an opposite role in inhibitory avoidance: whereas activation of the former receptors inhibits the acquisition of this response, activation of the latter facilitates it. Both 5-HT1A, 5-HT2A and 5-HT2C receptors seem to mediate the inhibitory action of serotonin on escape. PMID- 15342107 TI - PKC-alpha mediated alterations of indoleamine contents in diabetic rat brain. AB - We previously have reported that acute or chronic diabetes in animals resulted in altered neurotransmitter levels. In this study, we investigated the concentrations of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5 HIAA) in discrete areas of brain viz. striatum (ST), hippocampus (HC), hypothalamus (HT), midbrain (MB), pons medulla (PM), cerebellum (CB) and cerebral cortex (CCX) of control, untreated diabetic and insulin treated diabetic rats after 30 days. Alloxan (45 mg/kg) diabetic untreated rats, which showed hyperglycemia (>250 mg%), revealed significant increases of 5-HT level in ST, MB, PM, CB and CCX and the 5-HIAA level found to be increased significantly in ST, HC and MB. Whereas the insulin treated rats, which was maintained under normal glucose level (80-110 mg%), showed no significant changes in any of the areas studied. The expressions of PKC-alpha studied by immunoblotting also showed significant changes in ST, HC, MB, PM, CB and CCX that is identical to the changes of both 5-HT and 5-HIAA under similar condition, suggesting that the PKC alpha may regulate the synthesis and release of indoleamines in diabetic animals. PMID- 15342108 TI - Plant biopolyester cutin: a tough way to its chemical synthesis. AB - The chemical synthesis of an aliphatic biopolyester identical to the natural cutin which constitutes the major component of the cuticle of fruits and leaves of higher plants is for the first time achieved and reported. Potential applications of this new material is of great interest because its physical properties, non-toxicity, biodegradability, and availability of raw material. PMID- 15342109 TI - Effect of aluminium phosphide exposure on kinetic properties of cytochrome oxidase and mitochondrial energy metabolism in rat brain. AB - This study involves the effect of aluminium phosphide exposure on the kinetic characteristics of cytochrome oxidase and the mitochondrial respiratory chain function in rat brain. Mitochondrial preparations from both control and aluminium phosphide-treated rats demonstrated significant decrease in the maximal activity of cytochrome oxidase (approximately 50%) when expressed per unit membrane protein and on a turnover number basis (nmol/min/nmol haem a). The results indicated that there was a decrease in the catalytic efficiency of the active oxidase molecules on aluminium phosphide treatment. Arrhenius plot characteristics differ for cytochrome oxidase activity in mitochondria isolated from treated and control rats, in the break point of the biphasic plot which was shifted to a higher temperature. The decreased activity of cytochrome oxidase along with altered NADH and succinic dehydrogenase activities might have contributed towards a significant decline in state 3 and state 4 respiration. These alterations in the electron transport chain complexes in turn affected the ATP synthesis rate adversely in the mitochondria, isolated from treated rats. The data reflect the interaction of aluminium phosphide with redox chain components leading to the impairment of the electron transfer along the respiratory chain. PMID- 15342110 TI - Dynamics of sulfur amino acids in mammalian brain: assessment of the astrocytic neuronal cysteine interaction by a mathematical hybrid model. AB - A mathematically hybrid model was used to analyze three mechanisms by which cysteine could be produced in the brain to be used as preferential substrate in the synthesis of neuronal glutathione. In that way, the fluxes of sulfur compounds at the brain-blood barrier were integrated with their transport in astrocytes and neurons, and with their metabolism in astrocytes. We concluded that cysteine, in contrast with its precursor cystine, would not be taken up from the blood at the blood-brain barrier, but instead it must be lost continuously from astrocytes. Cysteine efflux is produced because the uptake of cystine in astrocytes is much greater than their cysteine demand to synthesize glutathione, hypotaurine and taurine. Once in the interstitial parenchyma, cysteine would be taken for the neurons, as backwardly by the endothelial cells. Remarkably, a close sulfur-macro balance can be maintained only if the surplus of the produced cysteine is transferred from the endothelial cells to the blood together with significant amounts of other sulfur-compounds, probably taurine and hypotaurine. In addition, the results obtained shown that alternative mechanisms of cysteine generation (i.e., nonenzymatic-thiol-disulfide exchange reaction, enzymatic cleavage of the glutathione effluxed from astrocytes) are not quantitatively significant under physiological conditions, in situ. PMID- 15342111 TI - Yeast growth selection system for the identification of cell-active inhibitors of beta-secretase. AB - Abeta peptides, which are believed to be at the origin of Alzheimer's disease (AD), are produced through the sequential processing of the transmembrane amyloid precursor protein (APP) by the beta- and gamma-secretase. The identification of small molecules that penetrate the brain and inhibit these secretases is of great therapeutic potential. Here, we describe a cellular selection system in yeast for the identification of inhibitors of the human beta-secretase BACE-1. Similar to the natural situation in mammalian cells, BACE-1 and its substrate APP are bound to membranes in secretory pathway compartments. Yeast cells expressing these human proteins have been engineered so as to grow under selective conditions only upon reduction of BACE-1 activity, thus allowing identification of compounds that, in addition to inhibiting BACE-1, must permeate cellular membranes and present no cytotoxic effects. Our results show that gradual reduction of BACE-1 expression in the engineered yeast strain resulted in gradual increase of cell growth rate. Moreover, two validated BACE-1 inhibitors, which have IC50 values between 7 and 8 microM in mammalian cell assays, stimulated yeast growth in a concentration-dependent manner. This effect was specific for BACE-1 since these compounds had no effect on yeast cells expressing a different secretase cleaving the APP substrate at the alpha-site. The target-specific cellular assay presented here is applicable in high-throughput screens for selecting inhibitors of defined secretases acting on natural substrates in a membrane-bound protein configuration. PMID- 15342112 TI - Unfolding and refolding of leucoagglutinin (PHA-L), an oligomeric lectin from kidney beans (Phaseolus vulgaris). AB - The unfolding and refolding of Phaseolus vulgaris Leucoagglutinin, a homotetrameric legume lectin, was studied at pH 2.5 and 7.2 using fluorescence, far- and near-UV circular dichroism (CD) spectroscopy, 8-anilino-1-naphthalene sulfonate (ANS) binding and FPLC techniques. This protein was found to refold even at pH 2.5 and also exhibited high refolding yield around 60% at pH 2.5 and 85% at pH 7.2. The refolding at pH 2.5 takes place with the formation of a dimeric intermediate. Although the hydrodynamic radius of the completely renatured protein and the dimer at pH 2.5 was found to be same, the ANS binding as well as far-UV CD spectra of the two were different. The denaturation kinetics at pH 2.5 followed single exponential pattern with the rate of denaturation being independent of protein concentration. The renaturation kinetics on the other hand was dependent on the protein concentration providing further evidence of an intermediate state during refolding. From these experiments the folding pathway of the protein at pH 2.5 was proposed. PMID- 15342113 TI - Respiratory nitrate reductase from haloarchaeon Haloferax mediterranei: biochemical and genetic analysis. AB - The Haloferax mediterranei nar operon has been sequenced and its regulation has been characterized at transcriptional level. The nar operon encodes seven open reading frames(ORFs) (ORF1 narB, narC, ORF4, narG, narH, ORF7 and narJ). ORF1, ORF4 and ORF7 are open reading frames with no assigned function, however the rest of them encoded different proteins. narB codes for a 219-amino-acid-residue iron Rieske protein. narC encodes a protein of 486 amino acid residues identified by databases searches as cytochrome-b (narC). The narG gene encodes a protein with 983 amino acid residues and is identified as a respiratory nitrate reductase catalytic subunit (narG). NarH protein has been identified as an electron transfer respiratory nitrate reductase subunit (narH). The last ORF encodes a chaperonin-like protein (narJ) of 242 amino acid residues. The respiratory nitrate reductase was purified 21-fold from H. mediterranei membranes. Based on SDS-PAGE and gel-filtration chromatography under native conditions, the enzyme complex consists of two subunits of 112 and 61 kDa. The optimum temperature for activity was 70 degrees C at 3.4 M NaCl and the stability did not show a direct dependence on salt concentration. Respiratory nitrate reductase showed maximum activity at pH 7.9 and pH 8.2 when assays were carried out at 40 and 60 degrees C, respectively. The absorption spectrum indicated that Nar contains Fe-S clusters. Reverse transcriptase (RT-PCR) shows that regulation of nar genes occurs at transcriptional level induced by oxygen-limiting conditions and the presence of nitrate. PMID- 15342114 TI - Evidence for differences in the metabolism of saxitoxin and C1+2 toxins in the freshwater cyanobacterium Cylindrospermopsis raciborskii T3. AB - The activity of paralytic shellfish poisoning (PSP) toxins biosynthetic enzymes was assayed in the cyanobacterium Cylindrospermopsis raciborskii T3 after inhibiting protein synthesis with chloramphenicol (CAM). The production of C1+2 and saxitoxin (STX) was sensitive to CAM with STX levels decreasing by 70% after 24-h exposure to the antibiotic. PSP toxin production was strongly promoted by arginine supplementation, with a maximum 476% increase in intracellular STX concentrations after 24-h exposure to 10 mM of the amino acid. However, arginine had no stimulating effect on PSP toxin levels if supplemented in combination with CAM at 10 microg l(-1). Addition of agmatine and proline to C. raciborskii T3 cultures in the presence of 10 microg l(-1) CAM increased C1+2 toxins levels, while having a negative or no effect on STX accumulation. In vitro, PSP toxin levels increased naturally in cyanobacterial extracts, with CAM and arginine having no influence on either C1+2 or STX synthesis. The evidence presented in this study suggests a possible difference between the metabolism of STX and the C1+2 toxins and indicated a high turnover rate of STX biosynthetic enzymes in C. raciborskii T3. PMID- 15342115 TI - Novel reactions of L-rhamnose isomerase from Pseudomonas stutzeri and its relation with D-xylose isomerase via substrate specificity. AB - Escherichia coli strain JM 109 harboring 6 x His-tag L-rhamnose isomerase (L-RhI) from Pseudomonas stutzeri allowed a 20-fold increase in the volumetric yield of soluble enzyme compared to the value for the intrinsic yield. Detailed studies on the substrate specificity of the purified His-tagged protein revealed that it catalyzed previously unknown common and rare aldo/ketotetrose, aldo/ketopentose, and aldo/ketohexose substrates in both D- and L-forms, for instance, erythrose, threose, xylose, lyxose, ribose, glucose, mannose, galactose, altrose, tagatose, sorbose, psicose, and fructose. Using a high enzyme-substrate ratio in extended reactions, the enzyme-catalyzed interconversion reactions from which two different products from one substrate were formed: L-lyxose, L-glucose, L tagatose and D-allose were isomerized to L-xylulose and L-xylose, L-fructose and L-mannose, L-galactose and L-talose, and D-psicose and D-altrose, in that order. Kinetic studies, however, showed that L-rhamnose with Km and Vmax values of 11 mM and 240 U/mg, respectively, was the most preferred substrate, followed by L mannose, L-lyxose, D-ribose, and D-allose. Based on the observed catalytic mode of action, these new findings reflected a hitherto undetected interrelation between L-RhI and D-xylose isomerase (D-XI). PMID- 15342116 TI - The expression of mitochondrial methylenetetrahydrofolate dehydrogenase cyclohydrolase supports a role in rapid cell growth. AB - Deletion of the gene encoding NAD-dependent methylenetetrahydrofolate dehydrogenase-cyclohydrolase (NMDMC) in mice was demonstrated previously to result in failure to establish definitive erythropoiesis in the developing liver. We examined the expression pattern of nmdmc to look for evidence that would support a tissue specific role for this activity. However, whole mount in situ hybridization revealed ubiquitous expression of nmdmc in the tissues of E9.5 and E10.5 embryos suggesting a broader role. Analysis of chimeras demonstrated that nmdmc-/- cells can survive in liver and other tissues of chimeras establishing that the null defect can be rescued by metabolites supplied by surrounding normal cells. Both the expression pattern and metabolite rescue support the proposal that mitochondrial NMDMC provides one-carbon units for purine synthesis during embryogenesis. The elevated expression of NMDMC in tumour cells, but not in surrounding normal cells, is predicted to result in significant differences in folate-mediated support for purine synthesis in the two cell types. PMID- 15342117 TI - Transglycosylation catalyzed by a Penicillium chrysogenum exo-1,5-alpha-L arabinanase. AB - Penicillium chrysogenum exo-arabinanase (Abnx), which releases arabinobiose from the nonreducing terminus of alpha-1,5-L-arabinan, was found to possess trans arabinobiosylation activity on various acceptors, such as aliphatic alcohols, sugars, and sugar alcohols. Abnx was found to prefer primary hydroxyl groups in polyhydric alcohols as acceptors over primary hydroxyl groups in monohydric alcohols. Among the 21 different compounds tested, glycerol was the best acceptor for the enzyme. The transfer product of glycerol was identified as O-alpha-L arabinosyl-(1-->5)-O-alpha-L-arabinosyl-(1-->1)-glycerol on the basis of the spectral data, fast atom bombardment-mass and 1H- and 13C-NMR. Unlike endo arabinanases, Abnx catalyzed the hydrolysis of linear arabinan without inverting the anomeric configuration. PMID- 15342118 TI - Complete RNAi rescue of neuronal degeneration in a constitutively active Drosophila TRP channel mutant. AB - RNA interference has been widely used to reduce the quantity of the proteins encoded by the targeted genes. A constitutively active, dominant allele of trp, TrpP365, causes massive degeneration of photoreceptors through a persistent and excessive Ca2+ influx. Here we show that a substantial reduction of the TRP channel protein by RNAi in TrpP365 heterozygotes completely rescues the neuronal degeneration and significantly improves the light-elicited responses of the eye. The reduction need not be complete, suggesting that rescue of degeneration may be possible with minimal side effects arising from overdepletion of the target protein. PMID- 15342119 TI - Inactivation of endopolyphosphatase gene PPN1 results in inhibition of expression of exopolyphosphatase PPX1 and high-molecular-mass exopolyphosphatase not encoded by PPX1 in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae possesses multiple forms of exopolyphosphatases, the enzymes involved in the metabolism of inorganic polyphosphates, which are important regulatory compounds. In S. cerevisiae, inactivation of endopolyphosphatase gene PPN1 leads to the inhibition of expression of both exopolyphosphatase PPX1 and high-molecular-mass exopolyphosphatase of approximately 1000 kDa not encoded by PPX1. In the single endopolyphosphatase mutant CRN, the expression of exopolyphosphatase PPX1 decreases 6.5-fold and 2.5 fold at the stationary and exponential growth phases, respectively, as compared with the parent strain CRY. In this mutant, the activity of the high-molecular mass exopolyphosphatase of approximately 1000 kDa decreases approximately 10-fold as compared with that in strains with the PPN1 gene. In a double mutant of PPX1 and PPN1, no exopolyphosphatase activity is detected in the cytosol at the stationary growth phase. Thus, the exopolyPase activity in cell cytosol depends on the endopolyPase gene PPN1. PMID- 15342120 TI - The CaPRP1 gene encoding a putative proline-rich glycoprotein is highly expressed in rapidly elongating early roots and leaves in hot pepper (Capsicum annuum L. cv. Pukang). AB - Most of the proline-rich cell wall glycoprotein genes isolated from higher plants are preferentially expressed in the transmitting tissues of the flower organ. In conducting expressed sequence tag (EST) analysis, which was prepared from 5-day old early roots of hot pepper (Capsicum annuum L. cv. Pukang), we identified a cDNA clone, pCaPRP1, encoding a putative cell wall proline-rich glycoprotein. CaPRP1 (Mr=28 kDa, pI=9.98) was most closely related to Nicotiana alata NaPRP4 (71%), while most distantly related to soybean PvPRP (37%). The predicted primary structure of CaPRP1 contains a putative N-terminal signal peptide, six repeats of the Lys-Pro-Pro tripeptide, four repeats of a five-amino acid sequence [Pro (Ser/The)-Pro-Pro-Pro] and one potential N-glycosylation site (Asn-Asn-Ser). In contrast to most proline-rich cell wall glycoprotein genes, CaPRP1 was highly expressed in rapidly elongating very early roots and young leaves as well as developing flower tissues. Although the physiological function of CaPRP1 is not yet clear, there are several possibilities for its role in cell expansion and elongation during early development of hot pepper plants. PMID- 15342121 TI - Fever, molecular excitability and epilepsy. PMID- 15342122 TI - Polymorphisms of casein kinase I gamma 2 gene associated with simple febrile seizures in Chinese Han population. AB - Casein kinase I gamma 2 isoform (CSNK1G2), a member of the large casein kinase I (CKI) family, may affect the development of brain, and associate with vesicular trafficking and neurotransmitter releasing from small synaptic vesicles. Based on our previous linkage analysis data that mapped our simple febrile seizures (FS) families to 19p13.3 and the function of CSNK1G2 in this region, CSNK1G2 was chosen as a candidate gene for FS. All of the 13 exons and their flanking introns of the CSNK1G2 gene were amplified and sequenced, and 10 single nucleotide polymorphisms (SNPs) were found. Using the three SNPs we found as markers, we conducted association studies in 60 FS patients and 101 normal controls. Allele and genotype frequencies of the SNPs IVS2-33C > T and 837C > T as well as the haplotype of the two SNPs were significantly different between FS patients and controls (P < 0.05). This study suggests that CSNK1G2 gene may be a susceptibility gene for FS in the northern Chinese Han population. PMID- 15342123 TI - Restraint stress affects hippocampal cell proliferation differently in rats and mice. AB - Granule cell neurogenesis occurs in the dentate gyrus of the mammalian hippocampus throughout adult life, and incorporation of bromodeoxyuridine (BrdU) into DNA can serve as a marker of cell division associated with such neurogenesis. We examined the effects of a stressor (3h of restraint) on hippocampal cell proliferation in Sprague-Dawley rats and C57BL/6J mice. Animals were killed immediately following restraint stress and their brains were prepared for immunohistochemical studies. Restraint stress caused similar significant increases in c-Fos immunoreactivity among cells in the hypothalamic paraventricular nucleus of both species, indicating that the stress experienced was similar. The restraint procedure also caused a significant decrease in BrdU labeling in the dentate gyrus of rats, as previously reported, but a significant increase in the same region in mice. Hippocampal cell proliferation appears to respond differently to restraint stress in these species. PMID- 15342124 TI - Scavenger receptor class B, type I mediates uptake of lipoprotein-associated phosphatidylcholine by primary porcine cerebrovascular endothelial cells. AB - We previously reported that scavenger receptor class B, type I (SR-BI) mediates uptake of lipoprotein-associated cholesteryl ester and Vitamin E by porcine brain capillary endothelial cells (pBCECs). In the present study we investigated whether SR-BI is capable of mediating phosphatidylcholine (PC) uptake by pBCECs from low- and high density lipoproteins. SR-BI-overexpressing CHO cells and pBCECs showed significantly enhanced uptake rates of PC from both lipoprotein classes. In addition, preincubation of pBCECs in the presence of both lipoprotein species resulted in a significant increase of the cellular fatty acid content, particularly linoleic and arachidonic acid. Our results suggest that uptake of lipoprotein-associated PC by the cerebrovasculature via SR-BI could generate a pool of lipids containing polyunsaturated fatty acids available for transport into deeper regions of the brain. PMID- 15342125 TI - Fulminant hepatic failure induced oxidative stress in nonsynaptic mitochondria of cerebral cortex in rats. AB - Fulminant hepatic failure (FHF) is a condition with sudden onset of necrosis of hepatocytes and degeneration of liver tissue without any established liver disease. FHF is associated with increased ammonia levels in blood and brain, which is supposed to be neurotoxic, ultimately leading to neuronal death. Evidences from previous studies suggest for mitochondrial dysfunctions under hyperammonemic conditions. In the present investigation, on thioacetamide-induced FHF rat models, studies were undertaken on cerebral nonsynaptic mitochondrial oxidative stress. The results of the present study reveal elevated lipid peroxidation along with reduced total thiol levels in the cerebral cortex mitochondria of experimental animals compared to saline treated control rats. In addition, the enzymatic activities of glutathione peroxidase and glutathione reductase were decreased, with an elevation in Mn-SOD activity. Overall, thioacetamide-induced FHF in rats enhanced the levels of lipid peroxidation coupled with impaired antioxidant defenses in the cerebral nonsynaptic mitochondria. PMID- 15342126 TI - FRAXE intermediate alleles are associated with Parkinson's disease. AB - There is evidence that male subjects with a clinical picture of action tremor, Parkinsonism, and cerebellar ataxia may have Fragile X premutations (FRAXA). We analyzed FRAXA and FRAXE triplet repeats in 203 male subjects with Parkinson's disease (PD) and 370 healthy controls. No full mutations or premutations at the FRAXA and FRAXE loci were found in the subjects with PD or in the controls. FRAXA allele distribution was similar in patients and controls. FRAXE intermediate alleles (31-60 repeats CCG) were found in 13 of 203 (6.4%) subjects with PD and in only one of the 370 (0.27%) healthy controls (P < 0.001), thus indicating that these relatively large alleles may be associated with PD. PMID- 15342127 TI - Projections from facial nucleus interneurons to the respiratory groups of brainstem in the rat. AB - Projections of the interneurons of the facial nucleus to respiration-related areas in the brainstem of the rat were revealed by unilateral, iontophorectic injection of the anterograde neuronal tracer, Phaseolus vulgaris leucoagglutinin (PHA-L), into the facial nucleus after motor neurons degeneration had been induced by axotomy of the facial nerve. Anterogradely labeled fibers and terminals were found bilaterally in the nucleus tractus solitarius, nucleus ambiguous and ventrolateral reticular formation in the medulla oblongata and ipsilaterally in the nucleus parabrachialis and nucleus Kolliker-Fuse in the pons. These results revealed that the interneurons in the facial nucleus have widespread projections to the respiratory groups in the brainstem, and suggest that the facial nucleus is involved not only in the control of muscles for facial expression but also in the regulation of functional respiratory activity. PMID- 15342128 TI - Nucleus accumbens single-unit activity in freely behaving male rats during approach to novel and non-novel estrus. AB - To assess the role of nucleus accumbens (NAcc) during approach to novel sexually arousing stimuli, we evaluated NAcc single-unit activity in male rats during the presentation of vaginal estrus extracted from novel and familiar female rats. After control stimulus presentations, animals were exposed to two estrous presentations from a female in the same or separate colony. A significantly larger proportion of units increased firing rate in the novel (11/32; 34%) than the familiar condition (2/28; 7%) during approach to the first but not the second presentation. Response magnitudes to novel but not familiar estrus were also greater than those during control trials. Collectively, these results provide further evidence of a role for the NAcc in the processing of novel sexually arousing information. PMID- 15342129 TI - Psychotic symptoms in Alzheimer's disease are not influenced by polymorphic variation at the dopamine receptor DRD3 gene. AB - It has been suggested that genetic influences unmasked during neurodevelopment to produce schizophrenia may appear throughout neurodegeneration to produce AD plus psychosis. Risk of schizophrenia and psychosis in Alzheimer's disease (AD) has been linked to polymorphic variation at the dopamine receptor DRD3 gene implying similar causative mechanisms. We tested this association in a large cohort of Alzheimer's disease patients with a diagnosis of probable AD of 3 years or more duration from the relatively genetically homogenous Northern Irish population. We assessed relationships between genotypes/alleles of the DRD3 BalI polymorphism and the presence or absence of psychotic symptoms (delusions, hallucinations) in AD patients during the month prior to interview and at any stage during the dementia. No significant associations were found when delusions and hallucinations were cross-tabulated against S and G alleles and SS, SG and GG genotypes. Logistic regression failed to detect any influence of APOE, gender, family history or prior psychiatric history. In conclusion, we were unable to confirm previously reported associations between the DRD3 BalI polymorphism and psychotic symptoms in AD. PMID- 15342130 TI - Spreading depression activates unfolded protein response. AB - Preconditioning is a process where a preceding non-lethal form of stress activates a stress response that protects cells against an otherwise lethal form of stress. Preconditioning can be induced in various ways including short-term ischemia or spreading depression. Here we investigated the effect of 1 h repetitive spreading depression on the unfolded protein response (UPR), a stress response activated under conditions associated with endoplasmic reticulum (ER) dysfunction. Spreading depression induced processing of xbp1 mRNA, indicative of an activation of UPR. Processing of xbp1 was paralleled by a rise in grp78 mRNA levels resulting from an activation of a signal transduction pathway that depends on protein synthesis. Preconditioning-induced activation of UPR may preserve ER functioning under pathological conditions interfering with ER functions. PMID- 15342131 TI - Association study of polymorphisms in the 5' upstream region of human DISC1 gene with schizophrenia. AB - Disrupted-in-Schizophrenia-1 (DISC1) is a gene in which a mutant truncation by a balanced t(1;11)(p42.1;q14.3) translocation is segregated with major psychiatric illness with a predominance of schizophrenic symptomatology in a large Scottish family. However, no functional polymorphisms have been detected that are associated with schizophrenia in general populations. As prior polymorphism searches in DISC1 have been focused on coding exons and flanking introns, the present study examined sequence variations in the 5' upstream region of DISC1. Screening of exon 1 through to approximately 1.0 kb upstream of exon 1 identified 6 polymorphisms, including 2 novel variants, -94C>A and -199(CG)(n). We tested these variants for associations with schizophrenia in the first set of a case (n = 198) and control (n = 198) panel, and found significant results with -274G>C (genotypic P = 0.01) and -215(TG)(n) (genotypic P = 0.039). However, we failed to replicate these associations in a second, larger independent patient (n = 532) and control (n = 519) sample. These results suggest that the genomic interval of DISC1 probably involved in transcriptional regulation does not display major genetic relevance in Japanese schizophrenia patients. PMID- 15342132 TI - Effect of sensory stimulus on striatal dopamine release in humans and cats: a [(11)C]raclopride PET study. AB - BACKGROUND: Sensory stimulation of the forelimb extremities constitutes a well established experimental model that has consistently shown to activate dopamine (DA) neurotransmission in the mammals' forebrain. OBJECTIVES: To visualize in vivo this modification of striatal DA release in healthy human volunteers using Positron Emission Tomography (PET) and [(11)C]raclopride. Experiments in humans were paralleled by experiments in anesthetized cats. Changes in endogenous DA release were assessed through its competition with [(11)C]raclopride binding (BP(raclo)), a radioligand probing DA D2-receptors. RESULTS: In humans no significant difference of BP(raclo) in caudate (with sensory stimulation: 2.0 +/- 0.3 versus without sensory stimulation: 2.2 +/- 0.3; P = 0.3) or putamen (2.6 +/- 0.3 versus 2.6 +/- 0.2; P = 0.9) ipsilateral to the stimulus was disclosed as a result of sensory stimulation. Similarly, no change of BP(raclo) was observed contralaterally to the stimulation in the caudate nucleus (with sensory stimulation: 2.0 +/- 0.4 versus without sensory stimulation: 2.1 +/- 0.2; P = 0.5) and the putamen (2.5 +/- 0.4 versus 2.6 +/- 0.2; P = 0.4). In cats the same results were obtained in the ipsilateral to stimulation striatum (with sensory stimulation: 2.5 +/- 0.03 versus without sensory stimulation: 2.4 +/- 0.05; P = 0.7). No change was also observed contralaterally to the stimulation (2.4 +/- 0.04 versus 2.5 +/- 0.06; P = 0.6). The [(11)C]raclopride binding remained unchanged by sensory stimuli in both humans and cats. CONCLUSION: This suggests that the DA release induced by sensory stimulus is mostly extrasynaptic whereas the synaptic DA release is probably small, which fits well with the absence of [(11)C]raclopride displacement. The mechanism of this extrasynaptic DA release could be related to a local action of glutamate on dopaminergic terminals via a thalamo-cortico-striatal loop. Present results also underline homology between cat and human responses to sensory stimuli and validate the use of cat brain to find physiological concepts in humans. PMID- 15342133 TI - Single and Sequential REM sleep episodes in humans: a phylogenetic left-over? AB - The occurrence of REM sleep in the rat appears to be under the control of either sleep related processes and homeostatic regulation of physiological variables. With respect to this, it has been observed that in this species REM sleep may occur in the form of two types of episodes, Single and Sequential episodes, which are supposed to play a different functional role. Since it is possible to distinguish Single and Sequential REM sleep episodes also in human beings, the aim of this pilot study was to asses whether a sleep deprivation may differently affect these two types of REM episodes. The sleep deprivation was induced in young human subjects by a progressive restriction of sleep within the same night period. Seventy-two PSG tracing belonging to six subjects have been analyzed. The results show that sleep deprivation does not significantly affect the relative occurrence of Single and Sequential REM sleep episodes, suggesting that in human beings these two types of REM episodes might not have a different functional role. PMID- 15342134 TI - Reduction of human neocortical and guinea pig CA1-neuron A-type currents by organic calcium channel blockers. AB - In epilepsy models, organic calcium antagonists regularly induce a transient activity increase before suppression of epileptiform discharges. This action was speculated to be mediated by a modulation of potassium currents. Since A-type currents potently regulate neuronal excitability, their modulation by calcium channel blockers was investigated in acutely isolated human neocortical temporal lobe neurons and CA1 neurons of guinea pigs using the whole-cell voltage-clamp technique. In human neurons, 40 microM nifedipine caused an amplitude reduction by 28% at a command potential of -6 mV and produced a biexponential, markedly accelerated current inactivation with time constants of 8.4 +/- 1.1 ms (n = 6) and 62.9 +/- 6.4 ms (n = 5). The time constant under control conditions was 50.1 +/- 8.5 ms (n = 6). Verapamil (40 microM) did not affect the current amplitude, but accelerated the monoexponential current inactivation from 40.2 +/- 7.1 ms to 13.3 +/- 0.8 ms (n = 9). Accordingly, verapamil accelerated the inactivation from 42.3 +/- 5.9 ms to 15.0 +/- 1.3 ms (n = 11) in guinea pig CA1 neurons, without affecting the current amplitude. In this preparation, it was shown that the two enantiomers of verapamil do not differ in their actions. The results show that the A-type current in human neocortical and in guinea pig hippocampal neurons is reduced by organic calcium channel blockers. PMID- 15342135 TI - Various immunization protocols for an acute motor axonal neuropathy rabbit model compared. AB - Various ganglioside immunization protocols were examined to refine the procedure for establishing an animal model of acute motor axonal neuropathy. The most effective was subcutaneous injection of an emulsion of 2.5mg of bovine brain ganglioside mixtures, keyhole lympet hemocyanin, and complete Freund's adjuvant to Japanese white rabbits, repeated at 3-week intervals. Under that protocol, all the rabbits developed marked flaccid paralysis associated with plasma anti-GM1 IgG antibody. This acute motor axonal neuropathy rabbit model also could be reproduced by the use of incomplete Freund's adjuvant, methylated bovine serum albumin, and New Zealand white rabbits. These results provide useful information for the confirmation of and further research on the model. PMID- 15342136 TI - Exogenous application of glucose induces aging in rat cerebral oligodendrocytes as revealed by alteration in telomere length. AB - To investigate aspects of aging on rat oligodendrocytes, cells of an oligodendrocyte cell line, so-called OLN-93, were cultured either in the presence or absence of glucose. Our data demonstrated that glucose-induced aging in vitro caused an elongation and thickening of cell processes and significantly increased the expression of netrin reflecting a more mature state of oligodendrocyte development. A possible age-inducing effect of glucose is also supported by the decrease of ras protein expression and shortening of telomeres in glucose-treated oligodendrocytes. The present study clearly shows that OLN-93 cells are an exciting and suitable model system for the investigation of age-inducing molecules and the analysis of signaling pathways involved in cerebral aging and degenerations. PMID- 15342137 TI - Prepro-orexin mRNA expression in the rat brain is increased during pregnancy. AB - The purpose of this study was to investigate whether orexin expression in the rat brain was changed during pregnancy. Brain samples were obtained from 5 nonpregnant rats and 10 pregnant rats (5; day 10 of gestation, and 5; day 20 of gestation). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed to investigate the expression of prepro-orexin mRNA and the housekeeping gene in the rat brain. The signals were quantified by the densitometric analysis. The distribution and expression of orexin-A and orexin-B were determined using immunohistochemistry. The ratio of the prepro-orexin mRNA expressions to the housekeeping gene expression in pregnant rat brain were significantly higher than that in nonpregnant control. There was no significant difference between prepro-orexin mRNA levels of day 10 and day 20 of gestation. Immunohistochemical staining for orexin-A and orexin-B was present in neurons within and around the lateral and posterior hypothalamic areas in both nonpregnant and pregnant rats. These results suggest that increased prepro-orexin mRNA levels at early gestational age in the maternal rat has a role on energy metabolism during pregnancy. PMID- 15342138 TI - Accuracy of a practicable EMG to force model for knee muscles. AB - A practicable EMG-force model is evaluated for muscles about the knee. The model included envelope signal processing and a gain-dependency of knee angle and angular velocity. Six healthy subjects participated in the experiments. For calibration, maximal isokinetic contractions about the knee were performed on a dynamometer with recordings of knee joint movement, net moment and EMG of thigh muscles. The model parameters were fitted on these calibration experiments. For validation, estimated moments from the EMG levels were compared to the actual exerted moments of two independent isokinetic contractions. Averaged RMS values of the difference ranged from 11 to 20% of the actual exerted moment. For isokinetic concentric contractions, the present model is suitable as a method to estimate muscle moments. PMID- 15342139 TI - Cannabinoids blocks tactile allodynia in diabetic mice without attenuation of its antinociceptive effect. AB - Diabetic neuropathic pain is one of the most commonly encountered neuropathic pain syndromes. However, the treatment of diabetic neuropathic pain is challenging because of partial effectiveness of currently available pain relievers. It is well known that diabetic animals are less sensitive to the analgesic effect of morphine, and opioids are found to be ineffective in the treatment of diabetic neuropathic pain. Cannabinoids are promising drugs and they share a similar pharmacological properties with opioids. It has been reported that cannabinoid analgesia remained intact and to be effective in some models of nerve injury. Thus, we investigated antinociceptive efficacy and the effects of cannabinoids on behavioral sign of diabetic neuropathic pain in diabetic mice by using WIN 55, 212-2, a cannabinoid receptor agonist. Diabetes was induced by streptozotocin (STZ) (200mg/kg) and animals were tested between 45 and 60 days after onset of diabetes. Antinociception was assessed using the radiant tail flick test. Mechanical and thermal sensitivities were measured by Von Frey filaments and hot-plate test, respectively. Tactile allodynia, but not thermal hyperalgesia developed in diabetic mice. Systemic WIN 55, 212-2 (1, 5 and 10mg/kg) produced a dose-dependent antinociception both in diabetic and control mice. WIN 55, 212-2-induced antinociception were found to be similar in diabetic mice when compared to controls suggesting efficacy of cannabinoid antinociception was not diminished in diabetic mice. WIN 55, 212-2 also produced a dose-dependent antiallodynic effect in diabetic mice. This study suggests that cannabinoids have a potential beneficial effect on experimental diabetic neuropathic pain. PMID- 15342140 TI - Estradiol reduces cytochrome c translocation and minimizes hippocampal damage caused by transient global ischemia in rat. AB - It is well-established that 17beta-estradiol (17beta-E(2)) confers neuroprotection to male and female rats exposed to focal cerebral ischemia, while less is known about the effects of the hormone under conditions of transient global ischemia. Since translocation of cytochrome c from the mitochondria to the cytosol is a critical step in apoptotic cell death after cerebral ischemia, we have investigated whether 17beta-E(2) interferes with such mechanism to exert neuroprotection. Global ischemia, induced in male Wistar rats by 5-min 4 vessel occlusion (4VO), resulted in a significant increase of cytosolic cytochrome c (cyt-c) levels as detected by Western blotting at 6h after reperfusion. 17beta E(2) (0.2mg/kg, i.p.) given 1h before ischemia minimized cytochrome c translocation and the latter effect was partially reversed by tamoxifen (0.25mg/kg, i.p.). Bilateral cell counting revealed that delayed hippocampal damage typically caused by 4VO was abolished by 17beta-E(2) and this was partially reversed by tamoxifen in the CA3 subregion, but not in CA1/CA2 or CA4. These findings provide the original observation that 17beta-E(2) reduces delayed hippocampal damage caused by 4VO in male rats and blocks cytochrome c translocation during the early stages of neuronal death, thus providing an important mechanism involved in estrogen-mediated neuroprotection. PMID- 15342141 TI - Cerebrovasodilation evoked by stimulation of subthalamic vasodilator area and hypoxia depends upon the integrity of cortical neurons in the rat. AB - In Sprague-Dawley rats symmetrical sites of the parietal cortex were microinjected with ibotenic acid (IBO, 10microg in 1microl) to lesion local neurons or with saline (1microl). Five days later, changes of cortical cerebral blood flow (CBF) in response to hypoxia and stimulation of the subthalamic vasodilator area (SVA) were measured using laser-Doppler flowmetry (LDF). The baseline CBF over the IBO- and saline-injected cortical sites did not differ significantly, but spontaneous waves of CBF were abolished over the lesioned sites. Elevations of CBF evoked by hypoxia or stimulation of SVA were attenuated by 54% and 88%, respectively (P < 0.05) over the lesioned sites, compared to saline-injected or non-injected sites. Hypercarbic cerebrovasodilation was comparable over all sites. We conclude that the SVA-evoked increase of CBF and about 50% of the hypoxia-evoked increase of CBF are mediated by excitation of cortical neurons. PMID- 15342142 TI - High voltage-activated Ca(2+) channel currents in isolectin B(4)-positive and negative small dorsal root ganglion neurons of rats. AB - Voltage-gated Ca(2+) channels in the primary sensory neurons are important for neurotransmitter release and regulation of nociceptive transmission. Although multiple classes of Ca(2+) channels are expressed in the dorsal root ganglion (DRG) neurons, little is known about the difference in the specific channel subtypes among the different types of DRG neurons. In this study, we determined the possible difference in high voltage-activated Ca(2+) channel currents between isolectin B(4) (IB(4))-positive and IB(4)-negative small-sized (15-30 microm) DRG neurons. Rat DRG neurons were acutely isolated and labeled with IB(4) conjugated to a fluorescent dye. Whole-cell patch clamp recordings of barium currents flowing through calcium channels were performed on neurons with and without IB(4). The peak current density of voltage-gated Ca(2+) currents was not significantly different between IB(4)-positive and IB(4)-negative neurons. Also, both nimodipine and omega-agatoxin IVA produced similar inhibitory effects on Ca(2+) currents in these two types of neurons. However, block of N-type Ca(2+) channels with omega-conotoxin GVIA produced a significantly greater reduction of Ca(2+) currents in IB(4)-positive than IB(4)-negative neurons. Furthermore, the IB(4)-positive neurons had a significantly smaller residual Ca(2+) currents than IB(4)-negative neurons. These data suggest that a higher density of N-type Ca(2+) channels is present in IB(4)-positive than IB(4)-negative small-sized DRG neurons. This differential expression of the subtypes of high voltage-activated Ca(2+) channels may contribute to the different function of these two classes of nociceptive neurons. PMID- 15342143 TI - Increase of galanin-like immunoreactivity in rat hypothalamic arcuate neurons after peripheral nerve injury. AB - Galanin and galanin receptors are widely distributed within the central nervous system, and may play important roles in pain signaling and modulation. In the present study, we examined the galanin immunoreactivity (IR) in the hypothalamus and the amygdala following peripheral nerve injury. Four weeks after the operation, the ipsilateral mechanical threshold in the spared nerve injury (SNI) group (0.87 +/- 0.33 g) was significantly lower than that in the sham group (12.53 +/- 3.41 g; P < 0.05). In the SNI group, the number of galanin-IR neurons per section in the arcuate nucleus (Arc) of the hypothalamus was 10.2 +/- 1.7, significantly higher than that in the sham group (5.6 +/- 1.0; P < 0.05). These data suggest that the galanin-ergic neurons in the Arc may be involved in the functional modulation of descending pain modulation system following peripheral nerve injury. PMID- 15342144 TI - Development of neuropathic pain is affected by bedding texture in two models of peripheral nerve injury in rats. AB - It has long been known that there are numerous genetic and environmental factors that affect the in vivo research of neuropathic pain. In this letter, we describe the impact that bedding material can have on the development of neuropathic pain behaviors in rodents. In two models of neuropathic pain, the Chronic Constriction Injury (CCI) and the partial axotomy, we demonstrated that features of the sawdust on which the animals are housed during experimentation have a clear effect on the development of mechanical hyperalgesia and chemical hypersensitivity. Rats housed on coarse sawdust presented with a much-reduced response to a pinprick and acetone test compared to counterparts housed on fine sawdust. It is therefore concluded that the development of specific stimulus modalities of neuropathic pain behavior following peripheral nerve injury can be influenced in part by environmental factors, in this case bedding texture. PMID- 15342145 TI - The role of nitrinergic connections in central cardiovascular responses mediated by physostigmine infused into posterior hypothalamus. AB - In the last few decades, cholinergic connections located into posterior hypothalamus (PH) have been implicated in the central regulation of blood pressure (BP). Here we investigated the role of nitric oxide (NO) in the blood pressure response elicited by infusion of physostigmine into PH of normotensive rats. In freely moving rats, physostigmine (60-200 nM) produced a dose- and time dependent elevation of BP which was antagonized by the antimuscarinic drug scopolamine (60 nM) and by L-NAME (100 microM), an inhibitor of NO synthase, both infused into the same site. In contrast, L-arginine (L-Arg; 100 microM), the precursor of NO, and glyceryltrinitrate (GTN; 140 nM), an NO donor, infused into the PH did not affect physostigmine-related pressor response. In rats pre-treated with Escherichia coli lipopolisaccharide (LPS; 0.5 microg i.p. 24h beforehand), however, scopolamine, L-Arg and GTN produced a decrease of BP, an effect antagonized by L-NAME. This suggests that NO only slightly modulates physostigmine-related pressor response elicited into PH of LPS-untreated rats. In contrast, the release of large amounts of NO generated by pre-treating rats with LPS, down-regulates cholinergic connections located at the PH, thus contributing in the central dysregulation of BP which can be found when high circulating endotoxin levels may occur. PMID- 15342146 TI - Spinal cholinergic involvement after treatment with aspirin and paracetamol in rats. AB - Aspirin and paracetamol have been shown to suppress non-inflammatory pain conditions like thermal, visceral and mechanical pain in mice and rats. The non inflammatory antinociception appears to be mediated by central receptor mechanisms, such as the cholinergic system. In this study, we tested the hypothesis that the non-inflammatory antinociception of aspirin and paracetamol could be mediated by an increase of intraspinal acetylcholine release. Microdialysis probes were placed intraspinally in anesthetized rats for acetylcholine sampling. Subcutaneously administered aspirin 100 and 300 mg/kg increased, while paracetamol 300 mg/kg decreased intraspinal acetylcholine release. Intraspinal drug administration did not affect acetylcholine release. Our results suggest that an increased intraspinal acetylcholine release could be involved in part of the non-inflammatory pain suppression by aspirin, but not by paracetamol. PMID- 15342147 TI - The effect of sagittal alignment on adjacent joint mobility after lumbar instrumentation--a biomechanical study of lumbar vertebrae in a porcine model. AB - BACKGROUND: The mechanisms and changes in range of motion of neighboring mobile segment (adjacent level) after the instrumented posterior stabilization are not completely understood. This study aims to investigate the effect of sagittal alignment on the adjacent joint mobility after lumbar instrumentation. METHODS: Eight fresh porcine lumbar spines were instrumented with pedicle screw implants from L2 to L4. Each specimen was tested in three different sagittal alignments. Group A were instrumented in lordotic alignment (lordosis 20 degrees ), Group B in straight alignment (lordosis 0 degrees ), and Group C in kyphotic alignment (kyphosis 20 degrees ). Hydraulic testing machine was used to generate an increasing moment in flexion and extension respectively for each specimen. The vertebral displacement of the disc between L1-L2 and L4-L5 were measured simultaneously with an extensometer. FINDINGS: There were no significant differences in vertebral displacement between the three different sagittal alignments in both the superior and inferior adjacent segments under extension motion. However, under flexion motion, the vertebral displacement on the superior adjacent segment (L1-L2) with kyphotic alignment was statistically larger than that of the straight and lordotic alignments (P = 0.0198 and P = 0.000473 respectively), and no differences were found between the three different sagittal alignments on the inferior adjacent segment (L4-L5). INTERPRETATION: The iatrogenically produced kyphotic lumbar spine by posterior instrumentation might cause larger adjacent joint mobility on the superior adjacent joint as compared to the instrumented lordotic lumbar spine. This study implies that an instrumented spine in lordosis is less likely to develop adjacent instability than a kyphotic spine. PMID- 15342148 TI - Flexion-relaxation response to cyclic lumbar flexion. AB - BACKGROUND: The epidemiology classify cyclic lumbar flexion as a risk factor for the development of cumulative low back disorder. Experimental biomechanical data confirming the epidemiology in humans are lacking. The purpose of this study, therefore, is to investigate the flexion relaxation response to sustained cyclic lumbar flexion in humans. METHODS: Twelve normal college aged males performed deep cyclic lumbar flexion at 0.1 Hz for 9 min while recording lumbar paraspinal electromyogram and kinematic data. FINDINGS: The most important observation of the study was the significant increase in the angular excursion of myoelectric silence during the deep part of the flexion phase. The observed increase in myoelectric silence consisted of earlier cessation of EMG during flexion and delayed activation of trunk extensors during extension. EMG magnitude, during flexion, increased approximately 30% over trials (P < 0.0001), and a smaller (10%), but, significant (P < 0.02) EMG magnitude increase was also observed during the extension phase of the cycle. Spasms, an indication of micro damage to viscoelastic tissues, were sporadic and appeared more frequently later in the session and mostly during the silent period. INTERPRETATIONS: It was concluded that increased myoelectric silence during prolonged cyclic flexion-extension demonstrates an enhanced flexion-relaxation phenomenon which reduces lumbar stability and may be detrimental to low back health. The presence of spasms confirm that sustained cyclic lumbar flexion results in micro damage in the viscoelastic tissues. Overall, a neuromuscular disorder was evoked due to a relatively short period of unloaded cyclic lumbar flexion. PMID- 15342149 TI - The effect of stabilization instruction on lumbar acceleration. AB - OBJECTIVE: To determine the effect of trunk stabilization instruction on lumbar acceleration. BACKGROUND: There is a paucity of information showing the impact of stabilization training and instruction on lumbar motion. Neuromuscular activation patterns have been implicated as an etiological factor in low back disorders. Knowledge of the short-term alterations in lumbar motion control, as assessed by acceleration, is an important first step to understanding the impact of trunk stabilization programs. METHODS: A calibrated, uniaxial accelerometer was positioned over the L4 spinous process of subjects (n =33) to record anterior/posterior acceleration during 10 repetitions of lower and upper limb movements (hip flexion/extension and biceps curl) before and after standardized stabilization instruction. RESULTS: Each repetition exhibited four stereotypical phases of acceleration. Instruction resulted in a substantial reduction in peak lumbar acceleration magnitude (P < 0.01) for all movement phases for both exercises (hip flexion 39.5%, biceps curl 30.8%). Lumbar peak accelerations for the biceps curl were substantially higher than peak accelerations for hip flexion (P < 0.05). CONCLUSIONS: This study demonstrated that stabilization instruction results in immediate alteration of kinematics (global reduction of limb movement induced lumbar acceleration) during upper and lower body movements. Reduced lumbar acceleration levels would arise from a change in the motor control strategy of trunk musculature due to instruction. PMID- 15342150 TI - Incremental and single trauma produce equivalent subfailure soft tissue injury of the cervical spine. AB - BACKGROUND: Automotive collision simulations have been performed using either incremental or single trauma. In single trauma, a single impact is performed, while in incremental trauma, a series of impacts of increasing severity are executed. Equivalency of incremental and single trauma for soft tissue injury severity due to the final impact has not been established. Thus, the purpose of the present study was to investigate whether incremental and single trauma produced similar cervical spine subfailure injury severity due to simulated frontal impacts. METHODS: Porcine cervical spine specimens (C2-T1) of the incremental trauma group were subjected to five frontal impacts (2, 3.5, 5, 6.5, 8 g), while single trauma specimens were subjected to a single impact (8 g). Flexibility tests were performed on specimens while intact and following each impact. Intact and post 8 g flexibility parameters were compared within incremental and single trauma groups and between groups. FINDINGS: No significant differences (P < 0.05) were found between incremental and single trauma groups when either intact or post 8 g flexibility parameters were compared. Significant increases in flexibility parameters from intact to post 8 g were observed in both groups, indicating soft tissue injury. INTERPRETATION: Incremental and single trauma produced equivalent subfailure cervical spine injury in simulated impacts, for the experimental conditions studied. This study may facilitate greater use of the incremental trauma protocol in future experimental designs. PMID- 15342151 TI - Isometric shoulder muscle activation patterns for 3-D planar forces: a methodology for musculo-skeletal model validation. AB - OBJECTIVE: To present an isometric method for validation of a shoulder model simulation by means of experimentally obtained electromyography and addressing all muscles active around the shoulder joints. BACKGROUND: Analysis of muscle force distribution in the shoulder by means of electromyography during motion tasks is hampered by artificial and non-linear amplitude modulation and is often limited to downward directed external forces. This application of EMG is therefore inadequate and insufficient for the validation of shoulder model simulations. We suggest an isometric method including multi-directional forces to overcome these problems. METHODS: A force with constant magnitude is actively rotated stepwise in 20 directions perpendicular around the arm while kept in one position. The isometric muscle activation (EMG) is a function of the clockwise rotated force angle, characterized by baseline activation, and a section of increased muscle activation characterized by baseline interception and direction and magnitude of maximum muscle activation. Comparison of the parameterized muscle activation with predicted muscle forces from model simulation illustrates the applicability for musculo-skeletal model validation. RESULTS: All recorded shoulder muscles were active over a section of force angles of at least 180 degrees. Some muscles demonstrated two activation sections. The estimated model sensitivity for the baseline interception was SD=5 degrees -10 degrees. The Principal Action was the most reliable parameter (SD=4 degrees ). A correlation of 0.778 was observed between model simulations and EMG recordings. CONCLUSIONS: The methodology addresses all shoulder muscles over a substantial section of planar force directions. This enables the comparison of experimentally determined direction of activation on- and offset and direction of maximum activation with equivalent muscle forces, predicted from model simulation. PMID- 15342152 TI - Active stability of the glenohumeral joint decreases in the apprehension position. AB - BACKGROUND: Muscle forces that compress the glenohumeral joint during mid-ranges of motion may lead to increased translational forces in end-range positions, such as the apprehension position, where symptoms of anterior instability occur. OBJECTIVE: The objective of this study was to quantify active stability provided by eight shoulder muscles in mid-range and end-range positions through muscle force vector analysis. METHODS: Lines of action were derived from a geometric model and muscle force magnitudes were estimated with electromyography-based techniques. Resultant muscle force vectors were calculated by summing individual muscle force vectors. RESULTS: Compared to mid-range positions, lines of action of resultant force vectors were more anteriorly directed in end-range positions compared to 15 degrees of abduction, up to 26 degrees. Consequently, anterior stability was lowest in the apprehension position. The magnitudes of the resultant force vectors were comparable to other studies. Based on a sensitivity analysis, lines of action of resultant force vectors vary up to 6 degrees within the population. CONCLUSIONS: Data obtained from this model will improve conservative management, post-surgical rehabilitation, and strength training protocols. PMID- 15342153 TI - Patterns of motion loss in subjects with idiopathic loss of shoulder range of motion. AB - BACKGROUND: Subjects with idiopathic loss of shoulder motion have difficulty sleeping, performing overhead activities, and completing activities of daily living. Treatment has been variable in approach and success. Cyriax's proposed shoulder capsular pattern was external rotation most limited followed by abduction followed by internal rotation. This investigation was performed to determine if a consistent pattern of motion loss was present in subjects with idiopathic loss of shoulder motion. METHODS: Repeated measurement of 25 (22 female) subjects' bilateral shoulder patterns of motion were compared. The three dimensional position of the scapula and humerus throughout active shoulder abduction, external rotation, flexion, internal rotation, and scapular plane abduction was evaluated with an electromagnetic motion capture system. Patterns of motion loss were determined for both shoulders. The proportions of peak shoulder motion to mean maximum non-involved shoulders abduction, external, and internal rotation were utilized to determine the pattern of motion loss. FINDINGS: There was a significant difference in motion loss patterns between the involved and non-involved shoulders with the arm at the side. The internal rotation less than abduction less than external rotation pattern was demonstrated in 14 of 25 (56%) involved shoulders. No pattern was present in 14 of 21 (67%) non-involved shoulders. With the arm abducted, internal rotation was the most limited motion in 23 of 25 (92%) involved shoulders. INTERPRETATION: The results did not support Cyriax's proposed glenohumeral capsular pattern. Defining the pattern of motion loss in subjects with idiopathic loss of shoulder motion may assist in diagnosis and treatment. PMID- 15342154 TI - Estimation of hip abduction moment based on body fixed sensors. AB - BACKGROUND: During standing and walking, abnormal trunk posture or movements may result from insufficient hip abductor force. Quantifying abduction moments requires inverse dynamics based on an appropriate model and measured data. Contrary to using a force plate and motion analysis system, body fixed sensors permit an analysis over subsequent strides and in a natural environment. Hence, the present paper analyses the feasibility of body fixed sensor-based estimations of the hip abduction moment. METHODS: Two models were defined to estimate the hip abduction moment based on data measured by accelerometers and gyroscopes; one model assumes a rigid trunk, the other assumes an upper and lower trunk segment. Data of five healthy subjects in different loading conditions while standing or walking were used to compare sensor-based estimations with estimations based on measured position and force data. FINDINGS: In different loading conditions, mean abduction moments calculated from position and force data varied between 0.27 and 0.62 Nm/kg. Generally, both sensor-based methods overestimated abduction moments. Results of a segmented trunk model showed best correspondence to the position and force-based method. Individual correlation coefficients between mean abduction moments in different loading conditions ranged from 0.72 to 0.98 (rigid trunk model) and 0.89 to 0.95 (segmented trunk model). INTERPRETATION: Hip abduction moments can be estimated based on the two models that use sensor data. Both methods are sensitive to different loading conditions. However, estimations are only valid when the trunk is supported by one leg alone, and particularly when the rigid trunk assumption is violated, the segmented trunk model yields better results. PMID- 15342155 TI - Sagittal plane biomechanics cannot injure the ACL during sidestep cutting. AB - BACKGROUND: Knee joint sagittal plane forces are a proposed mechanism of anterior cruciate ligament injury during sport movements such as sidestep cutting. Ligament force magnitudes for these movements however, remain unknown. The need to examine injury-causing events suggests elucidation via model-based investigations is possible. Using this approach, the current study determined whether sagittal plane knee loading during sidestep cutting could in isolation injure the anterior cruciate ligament. METHODS: Experiments were performed on subject-specific forward dynamic musculoskeletal models, generated from data obtained from 10 male and 10 female athletes. Models were optimized to simulate subject-specific cutting movements. Random perturbations (n=5000) were applied to initial contact conditions and quadriceps/hamstrings activation levels to simulate their effect on peak 3D knee loads. Injury via the sagittal plane mechanism was based on the criterion of an anterior drawer force greater than 2000 N. FINDINGS: Realistic neuromuscular perturbations produced significant increases in external knee anterior force and valgus and internal rotation moments. Peak anterior drawer force never exceeded 2000 N in any model, and thus failed to cause anterior cruciate ligament injuries. Valgus loads reached values that were high enough to rupture the ligament, occurring more frequently in females than in males. INTERPRETATION: Sagittal plane knee joint forces cannot rupture the anterior cruciate ligament during sidestep cutting. The interaction between muscle and joint mechanics and external ground reaction forces in this plane, places a ceiling on ligament loads. Valgus loading is a more likely injury mechanism, especially in females. Modifying sagittal plane biomechanics will thus unlikely contribute to the prevention of anterior cruciate ligament injuries. PMID- 15342156 TI - Effects of plantar fascia stiffness on the biomechanical responses of the ankle foot complex. AB - BACKGROUND: The plantar fascia is one of the major stabilizing structures of the longitudinal arch of human foot, especially during midstance of the gait cycle. Knowledge of its functional biomechanics is important for establishing the biomechanical rationale behind different rehabilitation, orthotic and surgical treatment of plantar fasciitis. This study aims at quantifying the biomechanical responses of the ankle-foot complex with different plantar fascia stiffness. METHODS: A geometrical detailed three-dimensional finite element model of the human foot and ankle, incorporating geometric and contact nonlinearities was constructed by 3D reconstruction of MR images. A sensitivity study was conducted to evaluate the effects of varying elastic modulus (0-700 MPa) of the plantar fascia on the stress/strain distribution of the bony, ligamentous and encapsulated soft tissue structures. FINDINGS: The results showed that decreasing the Young's modulus of plantar fascia would increase the strains of the long and short plantar and spring ligaments significantly. With zero fascia Young's modulus to simulate the plantar fascia release, there was a shift in peak von Mises stresses from the third to the second metatarsal bones and increased stresses at the plantar ligament attachment area of the cuboid bone. Decrease in arch height and midfoot pronation were predicted but did not lead to the total collapse of foot arch. INTERPRETATION: Surgical dissection of the plantar fascia may induce excessive strains or stresses in the ligamentous and bony structures. Surgical release of plantar fascia should be well-planned to minimise the effect on its structural integrity to reduce the risk of developing arch instability and subsequent painful foot syndrome. PMID- 15342157 TI - A biomechanical model of the effect of subtalar arthroereisis on the adult flexible flat foot. AB - OBJECTIVE: The hypothesis tested was that the increased load on the medial arch in the adult flat foot can be reduced through a 6 mm subtalar arthroereisis. DESIGN: A three-dimensional multisegment biomechanical model was used in conjunction with experimental data and data from the literature. BACKGROUND: Biomechanical models have been used to study the plantar fascia, medial arch height, subtalar motion, medial displacement calcaneal osteotomy and distribution of forces in the foot. METHODS: Responses of a normal foot, a flat foot, and a flat foot with a subtalar arthroereisis to an applied load of 683 N were analyzed and the distribution of support among the metatarsal heads and the moment about various joints were computed. RESULTS: The flattened foot results in an increase in the load on the head of the first metatarsal from 10% to 24% of the body weight, and an increase in the moment about the talo-navicular joint from 3.4 to 11.9 Nm. Insertion of a 6 mm cylinder into the sinus tarsi, subtalar arthroereisis, results in a shift of the load back toward the lateral column, decreasing the load on the first metatarsal to 6% of the body weight and decreasing the moment about the talo-navicular joint to 6.0 Nm. CONCLUSIONS: Our analysis indicates that a 6 mm subtalar arthroereisis in an adult flat foot model decreases the load on the medial arch. PMID- 15342158 TI - Biomechanical properties of skin in vitro for different expansion methods. AB - OBJECTIVE: In plastic surgery, clinicians are interested in replacing conventional expansion by rapid expansion, because the expansion period could be shortened greatly. Also people are concerned that skin properties after rapid expansion might not approach those after conventional expansion. DESIGN: Biomechanical testing of skin for different expansion methods. BACKGROUND: It would be useful to know how much the mechanical properties of skin have been altered during rapid and conventional expansion and during different maintaining times. METHODS: Tensile strength, stress-strain relationship, stress relaxation, and creep were measured by a material testing machine. RESULT: The biomechanical properties of experimental specimens differ significantly from those of their controls immediately after expansion, however, the difference is reduced with time. With the same maintaining period, the biomechanical properties of rapidly expanded skin are similar to conventionally expanded skin. CONCLUSION: Rapid skin expansion did not demonstrate any deleterious effect when compared with the conventional expansion. Extension of the maintaining period can improve the biomechanical properties of expanded skin. Therefore, rapid expansion with an extended maintaining period is acceptable in clinical practice. PMID- 15342159 TI - Modifying the gain of the visual feedback affects undisturbed upright stance control. AB - OBJECTIVE: To assess the effects of visual feedback gain, which express the amplitudes of the displacements of the centre of pressure displayed on a computer screen. DESIGN: The controlling variable, the centre of pressure trajectories, recorded using a force platform, were decomposed into two elementary motions: (1) the horizontal displacements of the centre of gravity and (2) the vertical projection of the difference between centre of pressure and the centre of gravity. These motions were processed through frequency analysis and modelled as fractional Brownian motion to assess their spatio-temporal linkage and their degree of control. BACKGROUND: Although tests to modify the feedback gain have already been carried out, the specific effects from a biomechanical and motor control point of view need to be assessed. METHODS: Thirteen healthy adults were tested through various visual feedback gains performed in random order. RESULTS: By increasing the visual feedback gain, no difference is observed between centre of pressure and centre of gravity motions whereas a progressive diminution of centre of gravity horizontal motions is seen. This latter effect is principally explained by a reinforcement of control during corrective processes. CONCLUSIONS: When the control of centre of gravity constitutes the main flaw in undisturbed stance maintenance, the efficiency of a visual feedback rehabilitation protocol should be largely improved by using an enhanced gain. PMID- 15342160 TI - Lateral ankle sprain: isokinetic test reliability and comparison between invertors and evertors. AB - OBJECTIVE: Evaluate reliability of isokinetic tests for inversion and eversion in individuals with recurrent lateral ankle sprain and compare inversion to eversion muscle performance in the same ankles. DESIGN: Reliability was evaluated by comparing the results of two ankle inversion/eversion isokinetic tests. Inversion and eversion were compared with the results presented by the tests. BACKGROUND: Previous studies showed reliable isokinetic tests for ankles with no sprain history. It is unknown if such tests can also be performed in lateral sprained ankles. It is unclear whether there is difference between invertors and evertors in these ankles. METHODS: Eleven men with recurrent ankle lateral sprain in one limb were tested using an isokinetic dynamometer. The tests consisted of 5 cycles of maximal contraction in 30 degrees /s and 120 degrees /s. Both injured and noninjured ankles were tested. Peak torque was recorded for analysis. RESULTS: The tests are reliable and intraclass coefficient correlation varied from 0.71 to 0.95. Invertors generated higher peak torques than evertors (P = 0.03) when injured ankles were tested at 120 degrees /s. CONCLUSIONS: Isokinetic inversion/eversion tests showed to be reliable. A decrease in peak torques can be seen during higher velocity tests. The only difference between invertors and evertors was seen when injured ankles were tested at 120 degrees /s. In this case invertors showed higher peak torques. PMID- 15342161 TI - Revascularisation for everyone? PMID- 15342162 TI - Transient myocardial ischaemia: new questions about autonomic responses. PMID- 15342163 TI - TIMPs, MMPs and cardiovascular disease. PMID- 15342164 TI - Can we quantify ischaemia during Dobutamine stress echocardiography in clinical practice? PMID- 15342165 TI - The slippery slope. PMID- 15342166 TI - The promise of myocardial repair--towards a better understanding. PMID- 15342167 TI - Natriuretic peptides in unstable coronary artery disease. AB - Patients with unstable coronary artery disease (CAD), i.e., unstable angina or non-ST-elevation myocardial infarction, vary widely in clinical presentation, prognosis and response to treatment. To select appropriate therapy, early risk stratification has become increasingly important. This review focuses on the emerging role of natriuretic peptides in the early assessment of patients with unstable CAD. We conclude that levels of brain natriuretic peptide (BNP) and N terminal pro-brain natriuretic peptide (NT-proBNP) are strongly associated to mortality and the risk of future congestive heart failure, and carry important prognostic information independent from previously known risk factors in unstable CAD. There are some data indicating that these markers can also be helpful in the selection of appropriate therapy in these patients but further studies are needed. Before a routine use of BNP or NT-proBNP in unstable CAD can be recommended, the cost-effectiveness of adding these new markers to the currently routine markers and their impact on selection of treatment needs further evaluation. PMID- 15342168 TI - Association of revascularisation with low mortality in non-ST elevation acute coronary syndrome, a report from GUSTO IV-ACS. AB - BACKGROUND: Immediate, as well as early, revascularisation is of benefit in patients with acute coronary syndromes (ACS) presenting with ST elevation. However, trials comparing invasive versus medical treatment in patients with an acute coronary syndrome without ST elevation do not consistently show improvement in survival after revascularisation. Accordingly, additional data are warranted. METHODS: The effect of revascularisation within 30 days on one-year survival in the GUSTO IV ACS trial was investigated. A total of 7800 patients were included with an acute coronary syndrome without ST elevation, documented by either elevated cardiac troponin or transient or persistent ST-segment depression. In this trial, comparing abciximab versus placebo as initial medical therapy, coronary angiography within 60 h after randomisation was discouraged. In 30-day survivors, those who underwent revascularisation were compared with 30-day survivors without revascularisation. Adjustments were made for patient characteristics, and for a propensity score that was adjusted for covariates associated with the likelihood of early revascularisation. FINDINGS: Of the 7496 patients who survived at least 30 days, 2265 (30%) underwent coronary revascularisation within 30 days: 789 patients CABG, 1450 PCI and 26 both CABG and PCI. Procedure-related mortality was low at 1.8%. Patients with revascularisation had a lower one-year mortality compared to medically treated patients (2.3% vs. 5.6%, p < 0.001). After multivariable analyses, patients with revascularisation had a relative risk of subsequent mortality within 1 year of 0.53 (95% CI 0.37-0.77) compared to patients without revascularisation. CONCLUSIONS: Revascularisation within 30 days is associated with an improved prognosis in ACS without ST-segment elevation. The relative high mortality in medically treated patients may be related in part to patient selection, but warrants further studies to improve outcome of these patients. PMID- 15342169 TI - Changes in autonomic nervous system activity: spontaneous versus balloon-induced myocardial ischaemia. AB - AIMS: Cardio-cardiac reflexes may be evoked by both myocardial ischaemia and coronary occlusion itself. The aim of the study was to assess the intrapatient behaviour of autonomic nervous system balance during spontaneous and balloon induced coronary ischaemia. METHODS AND RESULTS: We studied a group of patients admitted to the coronary care unit for acute coronary syndrome without ST-segment elevation who experienced spontaneous episodes of myocardial ischaemia during bed rest and ECG monitoring. The inclusion criterion was 80-90% lumen stenosis, amenable to angioplasty. Balloon coronary occlusion was performed at 4-6 atmospheres for 120 s. Autonomic nervous system activity was assessed by heart rate variability (HRV) analysis in frequency domain. We analysed 14 episodes of spontaneous ischaemia and 14 episodes of balloon coronary occlusion. During spontaneous ischaemia, HRV showed an increase in the low/high frequencies ratio (11.8 +/- 5.7), as compared to 5 min before and 5 min after (4.4 +/- 2.7 and 3.9 +/- 1.8, respectively) (p = 0.001). The opposite occurred during balloon coronary occlusion (0.8 +/- 0.4 vs. 3.9 +/- 2.0 and 5.1 +/- 2.1, respectively; p = 0.001). CONCLUSIONS: Balloon inflation and occlusion evokes baroreceptor vagal predominance in response to a stretch stimulus of the coronary artery. Conversely, spontaneous occlusion during unstable angina is accompanied by naturally occurring sympathetic activation. Sympathetic activation may have a role in the natural history of the disease. PMID- 15342170 TI - Relations of plasma total TIMP-1 levels to cardiovascular risk factors and echocardiographic measures: the Framingham heart study. AB - AIMS: Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a key regulator of extracellular matrix degradation. We examined relations of plasma total TIMP-1 to cardiovascular risk factors and echocardiographic left ventricular (LV) structure and function in a community-based sample. METHODS AND RESULTS: We studied 1069 Framingham Heart Study participants (mean age 56 years, 58% women) free of heart failure and previous myocardial infarction. Plasma TIMP-1 was higher in men compared with women, and increased with age, body mass index and total/HDL cholesterol ratio, but decreased with alcohol intake. Plasma TIMP-1 was also directly related to smoking, diabetes and use of anti-hypertensive treatment. Adjusting for age, sex and height, plasma TIMP-1 was positively associated with LV mass, wall thickness, relative wall thickness, end-systolic diameter, and left atrial diameter and the risk of having increased LV end-diastolic diameter or increased wall thickness, and negatively correlated with fractional shortening. Additional adjustment for clinical covariates attenuated the relations of plasma TIMP-1 to most echocardiographic measures. CONCLUSIONS: In our cross-sectional investigation, plasma total TIMP-1 was related to major cardiovascular risk factors and to indices of LV hypertrophy and systolic dysfunction. This raises the possibility that cardiovascular risk factors may influence cardiovascular remodelling via extracellular matrix degradation, which may be reflected in plasma TIMP-1 levels. PMID- 15342171 TI - Comparison of deformation imaging and velocity imaging for detecting regional inducible ischaemia during dobutamine stress echocardiography. AB - AIMS: To determine whether Doppler based myocardial tissue velocity imaging (TVI) or strain rate imaging (SRI) is more accurate in detecting stress-induced ischaemia during dobutamine stress echocardiography (DSE). METHODS AND RESULTS: Regional myocardial velocity, displacement, strain rate and strain patterns during DSE were investigated in 44 routine patients with known or suspected coronary artery disease. Simultaneous perfusion scintigraphy defined regional ischaemia. Curves and curved-M-mode patterns were analysed and receiver-operating characteristics of TVI and SRI parameters were compared by their area under the curve (AUC) in the receiver-operating-characteristics. In non-ischaemic segments, peak systolic velocity and strain rate increased significantly. Unlike SRI, TVI parameters had higher values in basal than in apical segments. In 47 segments of 19 segments DSE-induced ischaemia, which was proven by scintigraphy. In ischaemia, velocity and strain rate increased less. Post-systolic shortening (PSS) was always seen in SRI but not regularly in TVI. Peak systolic velocity and systolic displacement were the best TVI-parameters of stress-induced ischaemia (AUC 0.68 and 0.77, respectively.), in SRI it was the ratio of PSS and maximal segmental deformation (AUC=0.95, p < 0.0001). CONCLUSION: Compared to TVI, SRI parameters showed no major apico-basal gradient and had significantly higher diagnostic accuracy, comparable to conventional reading. SRI thus appears superior to TVI for regional ischaemia detection during DSE and may be preferred to support conventional DSE reading. PMID- 15342172 TI - Assessing myocardial perfusion with the transthoracic Doppler technique in patients with reperfused anterior myocardial infarction: comparison with angiographic, enzymatic and electrocardiographic indices. AB - AIMS: Doppler guidewire studies demonstrated that the no-reflow phenomenon in acute myocardial infarction is associated with characteristic coronary blood flow pattern. We investigated the potential of coronary flow measurement with transthoracic Doppler technique to detect the no-flow in the patients with reperfused infarction, and compared it to that of other modalities. METHODS AND RESULTS: We performed intracoronary myocardial contrast echocardiography after successful primary coronary intervention in the 94 patients with first, anterior wall infarction. Coronary blood flow in the left anterior descending artery was detected with transthoracic Doppler echocardiography within 24 h after reperfusion in 83 patients (88.3%). Twenty-two patients with the no-reflow had significantly lower systolic peak velocity (5.1 +/- 4.2 vs. 8.1 +/- 6.2 cm/s, p = 0.04), higher diastolic peak velocity (38.2 +/- 10.3 vs. 30.8 +/- 15.7 cm/s; p = 0.04), and shorter diastolic deceleration time (134 +/- 41 vs. 424 +/- 202 ms; p < 0.0001) than those with good-reflow. Systolic flow reversal was more frequently observed in those with no-reflow (18.2% vs. 3.3%, p = 0 .02). Diastolic deceleration time < 185 ms detected the no-reflow with far higher sensitivity/specificity (95.5%/95.1%) than TIMI frame count (45.5%/91.8%), ST resolution (54.5%/73.8%) and creatinine kinase-MB (54.5%/88.5%). CONCLUSION: Analysing coronary blood flow pattern can detect the no-reflow after anterior infarction better than other angiographic, electrocardiographic and enzymatic modalities. PMID- 15342173 TI - Role of beta1- and beta2-adrenoceptor polymorphisms in heart failure: a case control study. AB - BACKGROUND: We hypothesised that the polymorphisms of the genes encoding for beta1- and the beta2-adrenoceptors may have a role in the pathogenesis of heart failure (HF). We therefore compared the polymorphisms of the beta1-adrenoceptor gene (Arg389Gly), the beta2-adrenoceptor gene (Arg16Gly, Gln27Glu) and their combinations in patients with HF and normal subjects living in the same area. METHODS AND RESULTS: A total of 256 cases with HF (left ventricular ejection fraction < or = 40%) and 230 normal subjects were enrolled. The beta1- and beta2 adrenoceptor gene polymorphisms were assessed by PCR, followed by restriction enzyme digestion. No differences were observed in the distribution of any of the three genotypes studied in patients with HF and normal subjects. An analysis of the genotype combinations showed a non-significant increase in the risk of HF associated with the Arg389Gly16Gln27 (odds ratio = 1.4; 95%CI 0.5-3.6) and Arg389Gly16 Glu27 (odds ratio = 1.2; 95%CI, 0.5-2.8) homozygous allele combinations. CONCLUSION: None of the three most common polymorphisms of beta adrenoreceptors are associated with an increased risk of HF. PMID- 15342174 TI - Rate control is more cost-effective than rhythm control for patients with persistent atrial fibrillation--results from the RAte Control versus Electrical cardioversion (RACE) study. AB - Aims To evaluate costs between a rate and rhythm control strategy in persistent atrial fibrillation. Methods and results In a prospective substudy of RACE (Rate control versus electrical cardioversion for persistent atrial fibrillation) in 428 of the total 522 patients (206 rate control and 222 rhythm control), a cost minimisation and cost-effectiveness analysis was performed to assess cost effectiveness of the treatment strategies. After a mean follow-up of 2.3+/-0.6 years, the primary endpoint (cardiovascular morbidity and mortality) occurred in 17.5% (36/202) of the rate control patients and in 21.2% (47/222) of the rhythm control patients. Mean costs per patient under rate control were euro 7386 and euro 8284 under rhythm control. Cost-effectiveness analysis showed that per avoided endpoint under rate control, the cost savings were euro 24944. Under rhythm control, more costs were generated due to electrical cardioversions, hospital admissions and anti-arrhythmic medication. Costs were higher in older patients, patients with underlying heart disease, those who reached a primary endpoint and women. Heart rhythm at the end of study, did not influence costs. Conclusions Rate control is more cost-effective than rhythm control for treatment of persistent atrial fibrillation. Underlying heart disease but not heart rhythm largely accounts for costs. PMID- 15342175 TI - Embolic protection devices for carotid artery stenting: better results than stenting without protection? AB - AIMS: Carotid artery stenting (CAS) for carotid artery stenoses has become an alternative to carotid endarterectomy. However, CAS itself can cause cerebral ischaemic events. Embolic protection devices (PD) promise to reduce the incidence of these events. METHODS AND RESULTS: From July 1996 to March 2003, 1483 patients from 26 hospitals were included in the prospective CAS Registry of the ALKK study group. A PD was used in 668 of 1483 patients (45%). The use of a PD has grown rapidly over the years and reached 100% in 2003. Patients treated with a PD had prior carotid artery dilatation more often (3.5% versus 1%, p < 0.001), a prior myocardial infarction (34% versus 27.4%, p = 0.007) and a history of arterial hypertension (89.9% versus 78.6%, p = 0.007) compared to patients treated without a PD. A thrombus was more often visible in patients treated under distal protection (16.5% versus 8%, p < 0.001). The use of a PD led to a 10-min longer intervention (45 min versus 35 min median, p < 0.001). Patients treated with a PD had a lower rate of ipsilateral stroke (1.7% versus 4.1%, p = 0.007) and a lower rate of all non-fatal strokes and all deaths (2.1% versus 4.9%, p = 0.004) during the hospital stay. This was confirmed by multiple logistic regression analysis (adjusted OR = 0.45, 95% CI: 0.23-0.91, p = 0.026). A similar reduction could be found for symptomatic as well as asymptomatic carotid artery stenoses. CONCLUSION: Since 1996 there has been a steady increase in the use of PDs for CAS, with a 100% use in 2003. The use of a PD may lower the rate of ipsilateral strokes during CAS. PMID- 15342176 TI - Polymer blends used for the aqueous coating of solid dosage forms: importance of the type of plasticizer. AB - The aim of this study was to investigate the importance of the type of plasticizer in polymer blends used for the coating of solid dosage forms, comparing a lipophilic and a hydrophilic plasticizer (dibutyl sebacate (DBS) and triethyl citrate (TEC)). In vitro drug release from propranolol hydrochloride (propranolol HCl)-loaded pellets coated with blends of ethyl cellulose (EC) and Eudragit L (100:0, 75:25, 50:50, 25:75 and 0:100 w/w) was investigated at low as well as at high pH. To better understand the underlying mass transport mechanisms, the physicochemical properties of the film coatings (e.g. mechanical resistance, water uptake and dry weight loss behavior) were determined. Interestingly, drug release strongly depended on the type of plasticizer. Importantly, not only the slope but also the shape of the release curves was affected, indicating that the chemical nature of the plasticizer plays a major role for the underlying drug release mechanisms. Diffusion through the intact polymer coatings and/or through water-filled cracks was found to be dominating for the control of drug release. The relative importance of these pathways strongly depended on the polymer blend ratio and type of plasticizer. In contrast to DBS, TEC rapidly leached out of the coatings, resulting in decreasing mechanical resistances of the films and, thus, facilitated crack formation. In addition, the hydrophilicity of the plasticizer significantly affected the water uptake behavior of the film coatings and, hence, changes in the coatings' toughness and drug permeability. Also the relative affinity of the plasticizer to the different polymers was found to be of significance. In contrast to TEC, DBS has a higher affinity to EC than to Eudragit L, resulting in potential redistributions of this plasticizer within the polymeric systems and changes in the release profiles during storage. Importantly, these effects could be avoided with appropriate curing conditions and preparation techniques for the coating dispersions. PMID- 15342177 TI - Characterization of chitosan acetate as a binder for sustained release tablets. AB - A chitosan derivative as an acetate salt was successfully prepared by using a spray drying technique. Physicochemical characteristics and micromeritic properties of spray-dried chitosan acetate (SD-CSA) were studied as well as drug polymer and excipient-polymer interaction. SD-CSA was spherical agglomerates with rough surface and less than 75 microm in diameter. The salt was an amorphous solid with slight to moderate hygroscopicity. The results of Fourier transform infrared (FTIR) and solid-state (13)C NMR spectroscopy demonstrated the functional groups of an acetate salt in its molecular structure. DSC and TGA thermograms of SD-CSA as well as FTIR and NMR spectrum of the salt, heated at 120 degrees C for 12 h, revealed the evidence of the conversion of chitosan acetate molecular structure to N-acetylglucosamine at higher temperature. No interaction of SD-CSA with either drugs (salicylic acid and theophylline) or selected pharmaceutical excipients were observed in the study using DSC method. As a wet granulation binder, SD-CSA gave theophylline granules with good flowability (according to the value of angle of repose, Carr's index, and Hausner ratio) and an excellent compressibility profile comparable to a pharmaceutical binder, PVP K30. In vitro release study of theophylline from the tablets containing 3% w/w SD CSA as a binder demonstrated sustained drug release in all media. Cumulative drug released in 0.1 N HCl, pH 6.8 phosphate buffer and distilled water was nearly 100% within 6, 16 and 24 h, respectively. It was suggested that the simple incorporation of spray-dried chitosan acetate as a tablet binder could give rise to controlled drug delivery systems exhibiting sustained drug release. PMID- 15342178 TI - Receptor-mediated targeting of lipobeads bearing acetohydroxamic acid for eradication of Helicobacter pylori. AB - In the present context, phosphatidyl ethanolamine (PE) liposomes anchored polyvinyl alcohol (PVA) xerogel beads (lipobeads) bearing acetohydroxamic acid (AHA) was developed as a receptor-mediated drug delivery system for use in blocking adhesion of Helicobacter pylori and thereby preventing the sequelae of chronic gastric infections. PVA beads containing AHA were prepared by emulsification followed by low temperature crystallization method. Surface acylation with fatty acid chain was accomplished by treating PVA bare beads with palmitoyl chloride. The completion of this reaction was characterized by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) which confirmed the formation of an ester bond. Final formation of lipobeads was accomplished by combining acylated PVA beads with a PE liposome suspension. To confirm the specific binding propensity of lipobeads towards the PE specific surface receptors of H. pylori, we have performed in situ adherence assay and radiolabelling assay with human stomach cells and KATO-III cells, respectively. In both of these studies, pretreatment of H. pylori with lipobeads completely inhibited the adhesion of H. pylori to human stomach cells and KATO-III cells. These assays could serve as suitable in-vitro models for the study of binding efficacy of lipobeads with H. pylori surface receptors. In addition, the antimicrobial activity of the formulations was evaluated by growth inhibition (GI) studies with isolated H. pylori strain. The inhibitory efficacy of lipobeads was significantly higher compared to that of PVA bare beads. These results suggest that lipobeads could be a potential targeted drug delivery system in the treatment of H. pylori. PMID- 15342179 TI - Study of gamma-irradiation effects on aciclovir poly(D,L-lactic-co-glycolic) acid microspheres for intravitreal administration. AB - Gamma-irradiation effects on aciclovir poly(D,L-lactic-co-glycolic) acid (PLGA) microspheres, with gelatin as additive, were studied. Microspheres with a 2:2:10 aciclovir:gelatin:polymer ratio were prepared by the solvent evaporation method and sterilised by gamma-irradiation at a dose of 25 kGy. Loading efficiency, morphology (particle size analysis, scanning electron microscopy (SEM)), physical chemistry (infrared (IR) absorption spectrophotometry, differential scanning calorimetry (DSC), X-ray diffraction and gel permeation chromatography (GPC)) and in vitro release assays for 73 days were performed to evaluate the sterilisation effect on microsphere characteristics. After gamma-irradiation, no surface changes were observed by SEM. Microparticle mean diameter and aciclovir loading efficiency were not affected by gamma-ray exposition. IR spectroscopy, DSC and X ray diffraction showed no modification of the bulk properties of the microspheres or their components. The controlled release profiles of aciclovir-loaded microspheres for 73 days were not altered upon exposure to gamma-irradiation. GPC measurements showed a decrease in molecular weight of the polymer. The sterilisation method is adequate because microspheres underwent no change after exposition to gamma-irradiation. These favourable properties of the aciclovir loaded microspheres make them a suitable system for the intravitreal treatment of herpes virus infections, in an animal model. PMID- 15342180 TI - Skin penetration and distribution of polymeric nanoparticles. AB - Encapsulation using nanoparticulate systems is an increasingly implemented strategy in drug targeting and delivery. Such systems have also been proposed for topical administration to enhance percutaneous transport into and across the skin barrier. However, the mechanism by which such particulate formulations facilitate skin transport remains ambiguous. In this study, confocal laser scanning microscopy (CLSM) was used to visualize the distribution of non-biodegradable, fluorescent, polystyrene nanoparticles (diameters 20 and 200 nm) across porcine skin. The surface images revealed that (i) polystyrene nanoparticles accumulated preferentially in the follicular openings, (ii) this distribution increased in a time-dependent manner, and (iii) the follicular localization was favoured by the smaller particle size. Apart from follicular uptake, localization of nanoparticles in skin "furrows" was apparent from the surface images. However, cross-sectional images revealed that these non-follicular structures did not offer an alternative penetration pathway for the polymer vectors, whose transport was clearly impeded by the stratum corneum. PMID- 15342181 TI - Development of suppository formulation safely improving rectal absorption of rebamipide, a poorly absorbable drug, by utilizing sodium laurate and taurine. AB - To develop the safe formulation that can safely improve bioavailability of poorly absorbable drugs and that is practically available, we prepared the suppositories of rebamipide, a poorly soluble and poorly absorbable antiulcer drug, by employing the combinatorial use of sodium laurate (C12), an absorption enhancer, with taurine (Tau) or L-glutamine (L-Gln), an adjuvant exerting the cytoprotective action. Although the dissolution of rebamipide from fatty base (FB) suppository prepared using Witepsol H-15 was very slow, it was remarkably improved by the addition of C12 and L-Gln or Tau into the suppository. On the other hand, the dissolution of rebamipide from water-soluble base (WB) suppository prepared using polyethylene glycol was very rapid and the addition of adjuvants did not influence its dissolution so much. Rectal absorption of rebamipide examined in rats was remarkably improved by FB suppository containing C12 or both C12 and Tau, while the enhancing effect of C12 was relatively small in the case of WB suppositories. Biochemical and histopathological studies have confirmed that FB suppository containing both C12 and Tau or L-Gln did not cause any serious local damage, while FB suppository containing C12 only caused the erosion and shrinkage for a lot of rectal epithelial cells. In conclusion, FB suppository employing the combinatorial use of C12 with Tau could be a promising formulation that is effective and safe enough for poorly absorbable drugs to be practically administered. PMID- 15342182 TI - Development of mucoadhesive patches for buccal administration of ibuprofen. AB - A new formulation for topical administration of drugs in the oral cavity has been developed using several film-forming and mucoadhesive polymers. The films have been evaluated in terms of swelling, mucoadhesion and organoleptic characteristics. The best film, containing polyvinylpyrrolidone (PVP) as film forming polymer and carboxymethylcellulose sodium salt (NaCMC) as mucoadhesive polymer, was loaded with ibuprofen as a model compound and in vitro and in vivo release studies were performed. Statistical investigation of in vitro release revealed that the diffusion process was the main drug release mechanism and the Higuchi's model provided the best fit. In vivo studies showed the presence of ibuprofen in saliva (range 70-210 microg/ml) for 5 h and no irritation was observed. These mucoadhesive formulations offer many advantages in comparison to traditional treatments and can be proposed as a new therapeutic tool against dental and buccal diseases and disturbs. PMID- 15342183 TI - Poly(N-vinylpyrrolidone)-block-poly(D,L-lactide) as a new polymeric solubilizer for hydrophobic anticancer drugs: in vitro and in vivo evaluation. AB - The majority of novel anticancer drugs developed to date are intended for parenteral administration. Paradoxically, most of these drugs are water insoluble, delaying their clinical development. A common approach to confering water solubility to drugs is to use amphiphilic, solubilizing agents, such as polyethoxylated castor oil (e.g., Cremophor EL, CrmEL). However, these vehicles are themselves associated with a number of pharmacokinetic and pharmaceutical concerns. The present work is aimed at evaluating a novel polymeric solubilizer for anticancer drugs, i.e., poly(N-vinylpyrrolidone)-block-poly(D,L-lactide) (PVP b-PDLLA). This copolymer self-assembles in water to yield polymeric micelles (PM) that efficiently solubilize anticancer drugs, such as paclitaxel (PTX), docetaxel (DCTX), teniposide (TEN) and etoposide (ETO). A PM-PTX formulation was evaluated, both, in vitro on three different cancer cell lines and in vivo for its safety, pharmacokinetics, biodistribution and antitumor activity. In vitro, cytotoxicity studies revealed that the drug-loaded PM formulation was equipotent to the commercial PTX formulation (Taxol). In the absence of drug, PVP-b-PDLLA with 37% DLLA content was less cytotoxic than CrmEL. In vivo, acute toxicity was assessed in mice after a single injection of escalating dose levels of formulated PTX. PM PTX was well tolerated and the maximum tolerated dose (MTD) was not reached even at 100 mg/kg, whereas the MTD of Taxol was established at 20 mg/kg. At 60 mg/kg, PM-PTX demonstrated greater in vivo antitumor activity than Taxol injected at its MTD. Finally, it was shown in mice and rabbits that the areas under the plasma concentration-time curves were inversely related to PM drug loading. PMID- 15342184 TI - Effect of glycerol-L-lactide coating polymer on bone ingrowth of bFGF-coated hydroxyapatite implants. AB - Basic fibroblast growth factor (bFGF)-coated hydroxyapatite (HA) cylinders showed good bony incorporation in a previously conducted animal study. However, some cylinders exhibited focal inhomogeneous bone ingrowth. The purpose of the current study was to test whether glycerol-L-lactide polymer coating could improve release properties and bone incorporation of bFGF-coated HA implants. bFGF-coated HA cylinders with or without coating polymer were investigated for in vitro release of bFGF by an immuno-ligand-assay and also for bone ingrowth in miniature pigs after 42 and 84 days. Release from bFGF polymer composites was lower for the first 3 days compared to the other group but was more homogenous and detectable amounts were still found after 20 days. There was significant delay in bone ingrowth of the polymer implants in which even after 84 days bone ingrowth was not completed, whereas in the other group incorporation after 42 days occurred. Detailed histology revealed filling of the HA pores with the polymer, making ingrowth of the surrounding host bone impossible. Only after 84 days starting resorption of the polymer accompanied by bone ingrowth was found. The current study showed that glycerol-L-lactide is not suitable for coating of HA implants due to polymer induced "locking" of HA pores. PMID- 15342185 TI - The effect of penetration enhancers on drug delivery through skin: a QSAR study. AB - Skin penetration enhancers are used to allow formulation of transdermal delivery systems for drugs that are otherwise insufficiently skin-permeable. A full understanding of the mode of action could be beneficial for the design of potent enhancers and for the choice of the enhancer to be used in the topical formulation of a special drug. In this study, the structural requirements of penetration enhancers have been investigated using the Quantitative Structure Activity Relationship (QSAR) technique. Activities of naturally occurring terpenes, pyrrolidinone and N-acetylprolinate derivatives on the skin penetration of 5-fluorouracil, diclofenac sodium (DFS), hydrocortisone (HC), estradiol and benazepril have been considered. The resulting QSARs indicated that for 5 fluorouracil and diclofenac sodium, less hydrophobic enhancers were the most active. More precisely, molecular descriptors in the corresponding QSARs indicated the possible involvement of intermolecular electron donor-acceptor interactions. This was in contrast to the skin permeation promotion of hydrocortisone, estradiol and benazepril by enhancers, where a linear relationship between enhancement activity and n-octanol/water partition coefficients of enhancers was evident. The possible mechanisms of penetration enhancement as suggested by the QSARs will be discussed. PMID- 15342186 TI - Incorporation of different antibiotics into carbonated hydroxyapatite coatings on titanium implants, release and antibiotic efficacy. AB - Carbonated hydroxyapatite (CHA) coatings were applied onto titanium implants by using a biomimetic precipitation method. Different antibiotics were incorporated into the CHA coatings and their release and efficacy against bacteria growth were studied in vitro. The following antibiotics were used within this study: cephalothin, carbenicillin, amoxicillin, cefamandol, tobramycin, gentamicin and vancomycin. Increased concentrations of antibiotics in the coating solution led to a higher quantity of antibiotic incorporated into the CHA coating. Some antibiotics were better incorporated than others depending on their chemical structure. Antibiotics, containing carboxylic groups such as cephalothin, carbenicillin and cefamandol, were better incorporated than antibiotics lacking these groups. A bacterial inhibition test on Staphylococcus aureus bacteria showed inhibition of growth for all antibiotics that were released from the CHA coating. A release test was conducted in phosphate buffer saline PBS at pH 7.4 and 37 degrees C and showed that antibiotics containing carboxylic groups like cephalothin were slower released from the CHA coating than others. These results suggest that certain antibiotics are able to bind/chelate with calcium, resulting in a better incorporation into the CHA coating and a slower release. Antibiotics incorporated in CHA coatings on titanium implants might be used to prevent post surgical infections and to promote bone-bonding of orthopedic devices. PMID- 15342187 TI - A novel anionic dendrimer for improved cellular delivery of antisense oligonucleotides. AB - The optimal design of hybridisation-competent antisense oligonucleotides (ODNs) coupled with an efficient delivery system appear to be important prerequisites for the successful use of antisense reagents for gene silencing. We selected an antisense ODN complementary to an accessible region of the epidermal growth factor receptor (EGFR) mRNA with the aid of an antisense oligonucleotide scanning array. The scanning array comprised 2684 antisense ODN sequences targeting the first 120 nts in the coding region of EGFR mRNA. The array-designed antisense ODN was covalently conjugated to a novel anionic dendrimer using a pentaerythritol based phosphoroamidite synthon via automated DNA synthesis and the ability of this conjugate to effectively deliver and down-regulate EGFR expression in cancer cells was evaluated. Each dendrimeric structure had nine ODN molecules covalently linked to a common centre at their 3' termini. This dendrimer conjugate was markedly more stable to serum nucleases compared to the free ODNs and the cellular uptake of ODN-dendrimer conjugates was up to 100-fold greater as compared to mannitol, a marker for fluid phase endocytosis, and up to 4-fold greater than naked ODN in cancer cells. ODN-dendrimer uptake was energy-dependent and mediated, at least in part, via binding to cell surface proteins; a process that was inhibited by self-competition and by competition with free ODN, salmon sperm DNA, heparin and dextran sulphate. Fluorescent microscopy studies showed a combination of punctate and more diffuse cytosolic distribution pattern for fluorescently labelled ODN-dendrimer conjugate in A431 cells implying internalization by endocytosis followed by release and sequestration of the conjugate into the cytosol. Little or no conjugate appeared to be present in the nuclei of A431 cells. In vitro RNase H-mediated cleavage assays confirmed that covalently conjugated antisense ODNs in the dendrimer conjugate were able to hybridize and cleave the array-defined hybridisation target site within the EGFR mRNA without the need for ODN dissociation from the conjugate. In cell culture, ODN-dendrimer conjugates were effective in inhibiting cancer cell growth that correlated with a marked knockdown in EGFR protein expression. These data highlight a novel anionic dendrimer delivery system for gene silencing oligonucleotides that improved their biological stability, cellular delivery and antisense activity in cultured cancer cells. PMID- 15342188 TI - Hydrogel pullulan nanoparticles encapsulating pBUDLacZ plasmid as an efficient gene delivery carrier. AB - This study provides a method for enhancing the delivery of nucleic acid molecules to cells by encapsulating it inside the hydrogel pullulan nanoparticles. In this study, pullulan nanoparticles encapsulating pBUDLacZ plasmid has been prepared inside the aqueous droplets of w/o microemulsions. Transmission electron microscopy (TEM) image showed that the particles are spherical in shape with size of 45+/-0.80 nm diameter. Cell cytotoxicity studies as determined by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay demonstrated that cells incubated with nanoparticles remained more than 100% viable at nanoparticle concentration as high as 1000 microg/ml. From scanning electron microscope images, it was observed that the nanoparticles were internalised and the cells exhibited vacuoles in the cell body due to nanoparticle internalisation. Endocytosis of nanoparticles resulted in disruption of F-actin and beta-tubulin cytoskeleton of human fibroblasts. The efficacy of transfection in vitro on HEK293 and COS-7 cells demonstrated cell type dependence, with COS cells having a higher gene expression. The beta-gal expression in COS-7 cells by pullulan nanoparticle was comparable to commercially available Lipofectamine 2000. The results of this study are encouraging for the development of pullulan nanoparticles as an intracellular delivery system for drugs and genes. PMID- 15342189 TI - PEGylated polyethylenimine for in vivo local gene delivery based on lipiodolized emulsion system. AB - Polyethylenimine (PEI) is one of the most efficient vectors for non-viral gene delivery, whereas its poor transfection activity, compared to viral vectors, and cytotoxicity need to be improved for in vivo applications. In this study, we prepared two PEI conjugates with 6 and 10 wt.% of poly(ethylene glycol) (PEG) grafts (referred to PEI-PEG-6 and PEI-PEG-10, respectively) in order to investigate the effects of PEGylation on cytotoxicity and transfection activity in vitro. In addition, their suitability as vectors for local gene delivery in vivo was assessed by injecting lipiodolized emulsions containing polymer/DNA complexes into the femoral artery of Sprague-Dawley (SD) rats, occluded by a surgical suture to block inflow of the blood to the leg. Both PEGylated PEIs showed significantly lower cytotoxicity and higher transfection activity in COS-1 cells than PEI taken as a control; in particular, PEI-PEG-10 produced the most promising results. The stable water-in-oil emulsion, composed of aqueous domains containing the complexes and lipiodol as an oil phase, was formed in the presence of a hydrogenated castor oil. From in vivo experiments, it was found that all the complexes, dispersed in the lipiodolized emulsion, delivered effectively gene to muscle, surrounding the injection site, rather than other organs such as liver, spleen, kidney, heart and lung. The in vivo transfection activity of PEI-PEG-10 was 3-folds higher in muscle than that of PEI. Based on these results, it can be concluded that PEGylated PEIs (based on the lipiodolized emulsion system) hold a promising potential for local gene delivery in vivo. PMID- 15342190 TI - Dioleoyl phosphatidylethanolamine and PEG-lipid conjugates modify DNA delivery mediated by 1,4-dihydropyridine amphiphiles. AB - Complexes of amphiphilic cationic 1,4-dihydropyridines with DNA (lipoplexes) can be used for nonviral gene delivery. In order to achieve serum-resistant transfection system, DOPE and PEG-lipid conjugates were used to modify 1,4 dihydropyridine amphiphile DHP-12 complexes with DNA. The ability to bind DNA was examined by ethidium bromide displacement assay. Cellular uptake, transfection efficacy and intracellular trafficking of the lipoplexes were assessed using FACS, betagalactosidase gene transfection and confocal laser microscopy, respectively. Cytotoxicity was determined by MTT assay. DHP-12 lipoplexes that included DOPE showed enhanced cell uptake and transfection efficacy both in the absence and presence of serum. PEG-lipid conjugates, in contrast, impaired transfection. In conclusion, combination of DHP-12 with DOPE appears to be a promising transfection system. PMID- 15342191 TI - Reduced tissue immigration of monocytes by neuropeptide Y during endotoxemia is associated with Y2 receptor activation. AB - Neuropeptide Y (NPY) increases survival in experimental septic shock, which might be mediated by cardiovascular and/or immunological effects. To study the latter hypothesis, we monitored blood leukocyte subsets over 96 h after intravenous (i.v.) application of LPS in chronically i.v.-cannulated rats. LPS induced a dramatic leukopenia at 4 h after challenge, which was blunted in NPY-treated animals by stabilizing granulocyte and T-lymphocyte numbers. In addition, NPY treatment prevented tissue immigration of monocytes at early time points and consecutively mobilized activated monocytes from the third day after challenge. RT-PCR and in vitro adhesion studies provided evidence for a NPY Y2 receptor mediated effect on monocytes. Thus, NPY treatment has profound receptor-specific effects on the migration and adhesion of leukocytes under endotoxemic conditions. PMID- 15342192 TI - Inhibition of MMP-dependent chemotaxis and amelioration of experimental autoimmune uveitis with a selective metalloproteinase-2 and -9 inhibitor. AB - The chemotaxis of inflammatory cells depends on proteolytic disruption of extracellular matrix components. The metalloproteinases (MMP)-2 and -9 enable T lymphocytes to pass through basement membranes. Selective inhibition of only MMP 2 and -9 almost completely abolished the ability of lymphocytes to digest collagen. The chemotaxis of lymphocytes was reduced 40%. In our in-vivo model of experimental autoimmune uveitis (EAU), 46% of the animals in the treated group remained disease-free, whereas all animals in the control group developed EAU. The mean activity of the disease was also statistically significantly reduced. The data suggest that selective MMP-2 and -9 inhibition might be a treatment option. PMID- 15342193 TI - Mechanisms involved in prostaglandin E2-mediated neuroprotection against TNF alpha: possible involvement of multiple signal transduction and beta-catenin/T cell factor. AB - Cerebrospinal fluid prostaglandin E2 (PGE2) and tumor necrosis factor-alpha (TNF alpha) levels are elevated in patients with Alzheimer's disease (AD), which suggests that they are involved in neurodegeneration. We previously reported that TNF-alpha derived from human macrophages, in response to beta-amyloid or amyloidogenic C-terminal peptide, is a main mediator of inflammatory neurotoxicity. In a continuation of this work, the present study investigated the direct effect of PGE2, one of the major prostaglandins produced in the brain, on cell viability in SH-SY5Y neuronal cells treated with TNF-alpha. PGE2 did not promote neurotoxicity, but rather had a strong protective effect against TNF alpha by ameliorating TNF-alpha-induced apoptosis and also by rescuing the intracellular level of beta-catenin, a key transducer of the Wnt signaling pathway. PGE2-mediated stabilization of beta-catenin was accompanied by T-cell factor/lymphoid enhancer factor (Tcf/Lef)-mediated transcriptional activation, which was followed by an increase in the cyclinD1 level. Pharmacological studies provided further evidence supporting the notion that PGE2-mediated neuroprotection against TNF-alpha involves the stimulation of Tcf/Lef signaling through EP1-, EP2-, and EP4-mediated increases of beta-catenin in SH-SY5Y cells. In addition, this PGE2 effect appears to be dependent on the activation of protein kinase A, phosphatidylinositol 3-kinase, phospholipase C, and to a lesser extent protein kinase C. Thus, the molecular mechanism governing the inhibitory effect of PGE2 against TNF-alpha may involve the activation and cross talk of multiple signal transduction and play an important role in regulating the survival of neurons during the neurotoxic inflammatory response associated with neurodegenerative diseases including AD. PMID- 15342194 TI - ICAM-1 upregulation in the spinal cords of PLSJL mice with experimental allergic encephalomyelitis is dependent upon TNF-alpha production triggered by the loss of blood-brain barrier integrity. AB - Urate (UA) selectively scavenges peroxynitrite-dependent radicals and suppresses CNS inflammation through effects that are manifested at the blood-brain barrier (BBB). ICAM-1 upregulation in the spinal cord tissues of myelin basic protein (MBP) immunized mice is selectively inhibited by UA treatment. In contrast, the expression of ICAM-1 and other adhesion molecules by circulating cells is unchanged. Moreover, TNF-alpha expression in the CNS tissues of MBP-immunized mice is suppressed by UA treatment but TNF-alpha-induced ICAM-1 expression on neurovascular endothelial cells is not. Therefore the effect of UA on ICAM-1 upregulation in the CNS tissues is likely due to its known contribution to the maintenance of BBB integrity in MBP-immunized mice which in turn inhibits cell invasion into the CNS and prevents TNF-alpha production in CNS tissues. PMID- 15342195 TI - Interleukin-6 produces neuronal loss in developing cerebellar granule neuron cultures. AB - CNS levels of the cytokine interleukin-6 (IL-6) are elevated during CNS injury and disease, but it is unclear if IL-6 contributes to the pathologic process. Our studies show that in a well-characterized CNS developmental model system, primary cultures of rodent cerebellar granule neurons, chronic exposure to IL-6 during neuronal development can result in cell damage and death in a subpopulation of developing granule neurons. Chronic exposure to IL-6 also increased the susceptibility of the granule neurons to a toxic insult produced by excessive activation of NMDA receptors. These results are consistent with a role for IL-6 in the neuropathology observed in the developing CNS during injury and disease. PMID- 15342196 TI - Complement C3 and C5 play critical roles in traumatic brain cryoinjury: blocking effects on neutrophil extravasation by C5a receptor antagonist. AB - The role of complement components in traumatic brain injury is poorly understood. Here we show that secondary damage after acute cryoinjury is significantly reduced in C3-/- or C5-/- mice or in mice treated with C5a receptor antagonist peptides. Injury sizes and neutrophil extravasation were compared. While neutrophil density increased following traumatic brain injury in wild type (C57BL/6) mice, C3-deficient mice demonstrated lower neutrophil extravasation and injury sizes in the brain. RNase protection assay indicated that C3 contributes to the induction of brain inflammatory mediators, MIF, RANTES (CCL5) and MCP-1 (CCL2). Intracranial C3 injection induced neutrophil extravasation in injured brains of C3-/- mice suggesting locally produced C3 is important in brain inflammation. We show that neutrophil extravasation is significantly reduced in both C5-/- mice and C5a receptor antagonist treated cryoinjured mice suggesting that one of the possible mechanisms of C3 effect on neutrophil extravasation is mediated via downstream complement activation products such as C5a. Our data indicates that complement inhibitors may ameliorate traumatic brain injury. PMID- 15342197 TI - Chronically stimulated microglial cells do no longer alter their immune functions in response to the phagocytosis of apoptotic cells. AB - In an autoimmune inflammatory setting, ingestion of apoptotic T cells leads to a down-regulation of microglial immune functions. Recent studies have indicated that microglia can be matured by exposure to GM-CSF. GM-CSF stimulation led to a differentiated microglial phenotype and enhanced antigen-presenting capabilities. The secretion of TNF-alpha was significantly decreased by the uptake of apoptotic cells in unstimulated microglia, but not in GM-CSF-differentiated microglia. IL 10 secretion was unaffected. After ingestion of apoptotic cells, only previously unstimulated, but not GM-CSF-differentiated microglial cells decreased their T cell-activating potential as measured by IFN-gamma secretion in antigen-activated MBP-specific T cells. Thus, GM-CSF stimulation reduces the immunomodulatory functions of microglial cells. PMID- 15342198 TI - MHC and non-MHC gene regulation of disease susceptibility and disease course in experimental inflammatory peripheral neuropathy. AB - With a panel of rat strains, we demonstrate a strong impact of the MHC genotype on susceptibility and disease course in experimental autoimmune neuritis induced with peripheral nerve myelin or the P2 peptide 58-81 (KNTEISFKLGQEFEETTADNRKTK). Beside the MHC genotype, non-MHC genes determined disease susceptibility and resistance. The type of disease induced with P2 58-81 was strongly correlated to the strength of the MHC class II isotype interaction with P2 58-81. These findings suggest a link between susceptibility and acute versus chronic disease courses on one hand and the strength of the MHC class II molecule/peptide affinity on the other hand. PMID- 15342199 TI - Chronic electric stimulation of the midbrain ventral tegmental area increases spleen but not blood natural killer cell cytotoxicity in rats. AB - Previously we found that in conscious, freely behaving rats chronic electric stimulation of the lateral hypothalamus (LH) caused significant augmentation of natural killer cell cytotoxicity (NKCC) and a large granular lymphocyte (LGL) number more pronounced in the spleen than in the peripheral blood. The LH belongs to the so-called "brain reward system", a collection of the central structures whose activation produce positive emotions. The midbrain ventral tegmental area (VTA) is another prominent reward-relevant structure. In the present work, chronic electric stimulation of VTA (constant current 0.1 ms duration cathodal pulses delivered at frequency 50 Hz during 60 min daily session for 14 consecutive days) caused in rats an increase in the spleen but not in the peripheral blood NKCC (chromium release assay) without simultaneous effect on the number of large granular lymphocytes (LGL) (morphological method) and plasma level of prolactin (PRL), growth hormone (GH), corticosterone (COR), and testosterone (TST). This effect was anatomically specific as no influence of analogous thalamic stimulation on immune and endocrine response was found. The results obtained indicate that both reward-related areas VTA and LH enhance the cell-mediated immune response, represented by natural killer cytotoxicity, especially in the spleen. However, the effect pronounced by VTA is weaker than that of LH, possibly due to additional connections of LH with the hormonal and/or autonomic control systems. PMID- 15342200 TI - Expression of hemokinin 1 mRNA by murine dendritic cells. AB - Hemokinin 1 is encoded by preprotachykinin C (PPT-C) mRNA, and has been proposed as a regulator of B and T cell lymphopoiesis. Here we demonstrate the expression of mouse PPT-C mRNA by CD11b+ macrophages, CD11c+ dendritic cells and in the microglial cell line EOC 13.31. Expression was detected in freshly isolated CD11b+CD11c+ bone marrow cells, as well as in M-CSF expanded bone marrow-derived macrophages and GM-CSF expanded bone marrow-derived dendritic cells. There was preferential expression of PPT-C mRNA in dendritic cell subpopulations that were CD11b+, but not B220+ or GR-1+. These studies are the first to demonstrate PPT-C mRNA expression by cells of the myeloid lineage. PMID- 15342201 TI - Chronic restraint stress during early Theiler's virus infection exacerbates the subsequent demyelinating disease in SJL mice. AB - Chronic restraint stress, administered during early infection with Theiler's virus, was found to exacerbate the acute central nervous system (CNS) viral infection and the subsequent demyelinating phase of disease (an animal model of Multiple Sclerosis (MS)) in SJL male and female mice. During early infection, stressed mice displayed decreased body weights and spontaneous activity; while increased behavioral signs of illness and plasma corticosterone (CORT) levels. During the subsequent chronic demyelinating phase of disease, previously stressed mice had greater behavioral signs of the chronic phase, worsened rotarod performance, and increased inflammatory lesions of the spinal cord. In addition, mice developed autoantibodies to myelin basic protein (MBP), proteolipid protein peptide (PLP139-151), and myelin oligodendrocyte glycoprotein peptide (MOG33-55). PMID- 15342202 TI - BDNF mRNA expression in rat hippocampus following contextual learning is blocked by intrahippocampal IL-1beta administration. AB - The present study examined the modulating effects of an intrahippocampal injection of interleukin-1beta (IL-1beta) on brain-derived neurotrophic factor (BDNF) mRNA expression 0.5, 2, 4, and 6 h following contextual fear conditioning, a task known to increase BDNF mRNA, in rats. Contextual fear conditioning produced a time-dependent increase in BDNF mRNA that varied by region of hippocampus. IL-1beta blocked or reduced these increases in BDNF mRNA in the CA1, CA2, and dentate gyrus regions of the hippocampus, but had no effect in cortical regions. These data support the idea that IL-1beta-produced memory deficits may be mediated via BDNF mRNA reductions in hippocampus. PMID- 15342203 TI - Functional maturation of proteolipid protein(139-151)-specific Th1 cells in the central nervous system in experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is a widely adopted animal model system for studying human multiple sclerosis that affects the central nervous system (CNS). To understand the underlying pathogenic mechanisms of the autoimmune T cell response, localization, enumeration and characterization of autoreactive T cells are essential. We assessed encephalitogenic proteolipid protein epitope (PLP(139-151))-specific T cells in the periphery and CNS of SJL/J mice using MHC class II I-As multimers during both pre-clinical and clinical phases of PLP-induced EAE in conjunction with T cell function. Our results strongly suggest that PLP(139-151)-specific CD4+ T cells first expand primarily in the CNS-draining cervical lymph nodes and then migrate to the CNS. In the CNS, these PLP-specific CD4+ T cells accumulate, become activated and differentiate into effector cells that produce IFN-gamma in response to the self-peptide. PMID- 15342204 TI - Transcriptional pattern analysis of adrenergic immunoregulation in mice. Twelve hours norepinephrine treatment alters the expression of a set of genes involved in monocyte activation and leukocyte trafficking. AB - We investigated in vivo effects of norepinephrine (NE) on the transcription of 200 immunologically relevant genes in the mouse. Balb/c mice were s.c. implanted with NE containing retard tablets. Twelve hours later, splenic mRNA was prepared and hybridized onto cDNA microarrays containing the sequences of the major cytokines, their receptors and all CD-antigens of the mouse. Consistent results were obtained with a set of five genes: in the NE-treated animals four genes (CXCR4, VCAM1, IL-1R2, CD 14) were found 2-8 fold upregulated as compared to sham treated animals, whereas the gene for CCR3 was downregulated (< 0.5 fold). The findings were confirmed using quantitative reverse transcriptase Real Time PCR. These first results prove the usefulness of gene microarray technology towards transcription pattern analysis in neuroimmune interactions. Furthermore, they support the relevance of catecholamines in the regulation of leukocyte migration and the inflammatory response. PMID- 15342205 TI - Post-receptorial mechanisms underlie functional disregulation of beta2-adrenergic receptors in lymphocytes from Multiple Sclerosis patients. AB - Increased density of beta2-adrenergic receptors has been demonstrated on peripheral blood mononuclear cells (PBMCs) from Multiple Sclerosis (MS) patients. In this study we found that isoproterenol reduces T-cell proliferation and IFNgamma secretion in PBMCs cultures from healthy controls and IFNbeta-treated but not untreated MS patients. Reduced expression levels of G protein coupled receptor kinase (GRK)2/3 (p < 0.05) and increased isoproterenol-induced cAMP accumulation (p < 0.0001) were found in PBMCs from all MS patients. Dibutyryl cAMP reduced the proliferation of PBMCs from all subgroups but in a slighter manner in untreated MS patients. We conclude that signalling through beta2 adrenergic receptors is chronically up-regulated but functionally uncoupled to immunoregulatory functions of lymphocytes from MS patients. Disregulation downstream the cAMP-associated signalling may underlie such a phenomenon. PMID- 15342206 TI - Paraneoplastic antibodies detected by isoelectric focusing of cerebrospinal fluid and serum. AB - Patients with paraneoplastic neurological syndromes often produce intrathecal antibodies. We have employed isoelectric focusing and peroxidase-labeled anti-IgG or 35S-labeled Hu or Yo antigens to identify oligoclonal bands (OCB) representing either total IgG or Hu or Yo antibodies in serum and CSF of patients with paraneoplastic encephalomyelitis (PEM) or paraneoplastic cerebellar degeneration (PCD). OCBs representing paraneoplastic antibodies were found in all CSF, but in only three sera. Yo antibodies represented the majority of IgG bands in PCD-CSF, which may reflect a limited immune response, whereas in PEM/SN, there were numerous additonal IgG bands of unknown specificity, indicating a broader immune response in these patients. PMID- 15342207 TI - Osteopontin levels and increased disease activity in relapsing-remitting multiple sclerosis patients. AB - Osteopontin (OPN) has been identified as the most prominent cytokine-encoding gene expressed within multiple sclerosis (MS) lesions. Recently, we demonstrated that OPN plasma levels were elevated in active relapsing-remitting (RR) MS patients. In this longitudinal study, a trend was observed for OPN serum levels in relation to clinical exacerbations. Moreover, OPN protein levels were significantly elevated 1 month prior to increase of gadolinium (Gd)-enhancing lesion number, whereas no relation was observed between OPN levels and increase in Gd-enhancing lesion volume. Although no robust relation between OPN and disease activity was observed, these data suggest that OPN levels are elevated prior to increased disease activity in RR MS patients. PMID- 15342208 TI - Increased CXCL8 (IL-8) expression in Multiple Sclerosis. AB - Multiple Sclerosis (MS) is a chronic inflammatory disease of the CNS which is characterized by large mononuclear cell infiltration and significant demyelination. CXCL8 is a chemo-attractant for both neutrophils and monocytes and triggers their firm adhesion to endothelium. In this study, we demonstrate that serum CXCL8 and CXCL8 secretion from PBMCs are significantly higher in untreated MS patients compared to controls and are significantly reduced in MS patients receiving interferon-beta1a therapy. We suggest that CXCL8 may serve as a marker of monocyte activity in MS and may play a role in monocyte recruitment to the CNS. PMID- 15342209 TI - Interferon-beta enhances monocyte and dendritic cell expression of B7-H1 (PD-L1), a strong inhibitor of autologous T-cell activation: relevance for the immune modulatory effect in multiple sclerosis. AB - Antigen-presenting cells (APC) are considered to play a critical role in promoting the (re)activation of potentially autoreactive T cells in multiple sclerosis (MS), an inflammatory demyelinating disorder of the central nervous system (CNS). B7-H1 (PD-L1) is a novel member of the B7 family proteins which exert costimulatory and immune regulatory functions. Here we characterize the expression and functional activity of B7-H1 expressed on monocytes and dendritic cells (DC) of healthy donors and MS patients. B7-H1 is constitutively expressed on monocytes and differentially matured DC, but not on B cells. IFN-beta, the principle immune modulatory agent used for the treatment of MS, strongly enhances B7-H1 expression on monocytes and semi-matured DC, but not B cells, in vitro. Importantly, B7-H1 expressed on APC strongly inhibits autologous CD4 T-cell activation. Neutralization of B7-H1 on monocytes or differentially matured monocyte-derived DC markedly increases the secretion of the pro-inflammatory cytokines, IFN-gamma and IL-2, T-cell proliferation, and the expression of T-cell activation markers. B7-H1 exhibits strong inhibitory effects when expressed on monocytes, immature or semi-mature DC, but less so when expressed on fully matured DC. B7-H1-dependent immune inhibition is in part mediated by CD4/CD25+ regulatory T cells. There is no difference in the baseline expression levels of monocytic B7-H1 between untreated MS patients and healthy donors. However, both groups show a significant concentration-dependent up-regulation of B7-H1 mRNA and protein in response to IFN-beta in vitro. Serial measurements of B7-H1 mRNA in MS patients before and 6 months after initiation of IFN-beta therapy corroborated the relevance of these results in vivo: Nine of nine patients showed a significant increase in B7-H1 mRNA levels after 6 months of IFN-beta therapy (median 1.04 vs. 8.78; p<0.05, two-sided t-test). Accordingly, protein expression of B7-H1 on monocytes was up-regulated after 24 h of IFN-beta application. In summary, B7-H1 expressed on APC acts as a strong inhibitor of autologous CD4 T cell activation and may thus contribute to the maintenance of peripheral immune tolerance. IFN-beta up-regulates B7-H1 in vitro and in MS patients in vivo and might represent a novel mechanism how IFN-beta acts as a negative modulator on APC T-cell interactions in the periphery. PMID- 15342210 TI - An open-label, comparative study of the efficacy and safety of once-daily dose of enoxaparin versus unfractionated heparin in the treatment of proximal lower limb deep-vein thrombosis. AB - BACKGROUND: Treatment of deep-vein thrombosis (DVT) with a once-daily regimen of enoxaparin, rather than a continuous infusion of unfractionated heparin (UFH) is more convenient and allows for home care in some patients. This study was designed to compare the efficacy and safety of these two regimens for the treatment of patients with proximal lower limb DVT. METHODS: 201 patients with proximal lower limb DVT from 13 centers in Brazil were randomized in an open manner to receive either enoxaparin [1.5 mg/kg subcutaneous (s.c.) OD] or intravenous (i.v.) UFH (adjusted to aPTT 1.5-2.5 times control) for 5-10 days. All patients also received warfarin (INR 2-3) for at least 3 months. The primary efficacy endpoint was recurrent DVT (confirmed by venography or ultrasonography), and safety endpoints included bleeding and serious adverse events. The rate of pulmonary embolism (PE) was also collected. Hospitalization was at the physician's discretion. RESULTS: Baseline patient characteristics were comparable between groups. The duration of hospital stay was significantly shorter with enoxaparin than with UFH (3 versus 7 days). In addition, 36% of patients receiving enoxaparin did not need to be hospitalized, whereas all of the patients receiving UFH were hospitalized. The treatment duration was slightly longer with enoxaparin (8 versus 7 days). There was a nonsignificant trend toward a reduction in the rate of recurrent DVT with enoxaparin versus UFH, and similar safety. CONCLUSIONS: A once-daily regimen of enoxaparin 1.5 mg/kg subcutaneous is at least as effective and safe as conventional treatment with a continuous intravenous infusion of UFH. However, the once daily enoxaparin regimen is easier to administer (subcutaneous versus intravenous), does not require aPTT monitoring, and leads to both a reduced number of hospital admissions and an average 4-day-shorter hospital stay. PMID- 15342211 TI - Plasma levels of thrombin activatable fibrinolysis inhibitor in normal and preeclamptic pregnant women. AB - To explain the pathogenesis of preeclampsia with coagulation induction or a defective fibrinolysis, various hemostatic parameters were studied and different treatment modalities targeting these parameters were evaluated. Considering the role of TAFI in down-regulation of fibrinolysis, in our study we have investigated whether TAFI contributes to impaired fibrinolysis in patient with preeclampsia. Thirty patients with preeclampsia (mean age +/- SD 25.7 +/- 4.53; range 17-36) and 30 normal pregnant women as control group (mean age +/- SD 28 +/ 5.26; range 21-38) were included in our study. TAFI antigen was determined using an ELISA kit for quantitative measurement. The mean TAFI antigen levels were 12.55 +/- 1.88 microg/ml in patients with preeclampsia and 12.29 +/- 3.0 microg/ml in normal pregnant women. A statistically significant difference was not found between TAFI antigen levels of two groups (p > 0.05). In order to clarify the role of TAFI in the pathogenesis of preeclampsia, in addition to plasma TAFI levels, its synthesis, activation and metabolism should also be evaluated. PMID- 15342212 TI - Patient preferences for capillary vs. venous INR determination in an anticoagulation clinic: a randomized controlled trial. AB - BACKGROUND: Patients who are receiving warfarin therapy require frequent blood testing to monitor the intensity of anticoagulation. Although previous studies suggest that capillary blood monitoring of the international normalize ratio (INR) is rapid and reliable, patient preferences for the method of blood drawing have not been investigated. METHODS: We performed a randomized controlled trial of patients attending an anticoagulation clinic in which patients were randomly allocated to undergo capillary or venous INR monitoring. Patient satisfaction with the outpatient visit, pain associated with blood drawing, and time spent in the clinic were assessed for each patient. RESULTS: Sixty patients were studied. Using a 10-point visual analogue scale to quantify patient satisfaction (0-very satisfied; 10-very dissatisfied), patients expressed a strong preference for capillary INR monitoring over venous INR monitoring (1.64 vs. 4.45; P < 0.001). Using a 10-point visual analogue scale to quantify pain with blood sampling (0-no pain; 10-very painful), patients who underwent capillary INR testing had less pain than venous INR testing (0.83 vs. 2.23; P < or = 0.004). Patients spent, on average, 33 fewer minutes in the clinic with capillary INR testing than venous INR testing (P < 0.001). DISCUSSION: Our findings support the routine use of capillary blood testing, using a portable monitor, for the management of patients in outpatient anticoagulation clinics. PMID- 15342213 TI - Release of soluble CD40 ligand after platelet activation: studies on the solubilization phase. AB - sCD40L is released from platelets as a soluble, proteolyzed form of CD40 ligand (CD40L; CD154) which is exposed on the surface after platelet activation. Ethylenediaminetetraacetate (EDTA), the CD40-blocking antibody G28-5, and GPIIb IIIa antagonists are known to inhibit the solubilization when added prior to activation. It is assumed that the surface expression of CD40L is a result of a separate fast process and that the solubilization is secondary to this. The release of sCD40L in this solubilization phase has been studied; that is, inhibitory substances were added to platelet-rich plasma (PRP) 10 min after addition of the activation agonist (100 microM SFLLRN), at which time the secretion phase was over as tested with beta-thromboglobulin (beta-TG). G28-5 (10 microg/ml) and EDTA (5 mM) inhibited the solubilization phase which did not require the presence of an activation agonist. Prostaglandin E1 (PGE1; 20 microM) and cytochalasin D (C8273; 60 and 100 microM), which exert their effects intracellularly, inhibited the solubilization even in the presence of abciximab (ReoPro; 40 microg/ml). The intracellular effect was not related to CD40L containing microparticles as demonstrated by ultracentrifugation. Intracellular alkalinization by preincubation of PRP with 20 mM NH4Cl for 60 min resulted in a small but reproducible reduction in the amount of extracellular sCD40L. SFLLRN induced solubilization of CD40L also from the platelets of a Glanzmann's thrombasthenia patient lacking GPIIb-IIIa, albeit at a lower rate than from normal platelets, and fibrinogen enhanced the solubilization from washed normal platelets. The data show that the solubilization of CD40L not only depends on reactions on the platelet surface but also that intracellular structures are engaged even during the solubilization phase. PMID- 15342214 TI - Methyl-methacrylate bone cement surface does not promote platelet aggregation or plasma coagulation in vitro. AB - Leakage of viscous bone cement into venous blood possibly resulting in pulmonary embolism may occur during percutaneous vertebroplasty. Our aim was to study if bone cement surface or cement liquid component could induce platelet aggregation or plasma coagulation in vitro. Two types of commonly used methyl-methacrylate bone cement, Palacos (Heraeus Kulzer, Germany) and Vertebroplastic (DePuy, Acro Med, England), were smeared on thin glass slides that were inserted over the bottom of cuvettes immediately or after 24 h, and platelet aggregation was recorded over 10 min. Bone cement liquid component, containing methyl methacrylate monomer and N,N-dimethyl-p-toluidine, was tested in 2% and 4% final concentration. Partial thromboplastin time (PTT) was determined by the hook method in the presence of bone cement-smeared glass slides or 6% bone cement liquid. Both types of bone cement, either fresh or aged, did not promote platelet aggregation, whereas collagen-coated glass slides induced substantial platelet aggregation (65 +/- 37%). On the other hand, bone cement liquids reduced platelet aggregation induced by collagen solution to an average of less than 15% (p < 0.01). Bone cement, fresh or aged, had no effect on PTT, but bone cement liquids significantly prolonged PTT: median and 1st-3rd interquartile range 149 (96-171) s for Vertebroplastic and 132 (99-194) s for Palacos, p = 0.03 for both comparisons with normal pool plasma without additives that had PTT of 69 (62-71) s. We conclude that the surface of fresh or aged bone cement is not thrombogenic in vitro. The bone cement liquid inhibits platelet aggregation and plasma clotting in relatively high concentrations that cannot be expected in vivo. PMID- 15342215 TI - Inhibition of thrombin-induced signaling by resveratrol and quercetin: effects on adenosine nucleotide metabolism in endothelial cells and platelet-neutrophil interactions. AB - BACKGROUND: Thrombin downregulates endothelial ectonucleotidase activity resulting in high levels of adenosine diphosphate (ADP) and adenosine triphosphate (ATP) which lead to platelet, leukocyte and endothelial activation. Depending on adenosine nucleotide levels, resting platelets inhibit and thrombin activated platelets increase respiratory burst of neutrophils. Whether the red wine polyphenols quercetin and resveratrol affect thrombin-dependent adenosine nucleotide, metabolism and thrombin-induced signaling is unknown. MATERIALS AND METHODS: ATP and ADP secretion by platelets, the impact on subsequent oxidative burst activity in neutrophils and CD39/ATPdase function in endothelial cells (ECs)was studied. Cell migration was measured in modified Boyden chambers; adenosine metabolites were quantified by high-performance liquid chromatography (HPLC). Signal transduction was studied by Western blotting. RESULTS: Quercetin and resveratrol inhibited thrombin-induced ADP and ATP secretion from platelets in a concentration-dependent manner. Augmented respiratory burst of neutrophils in response to thrombin-activated platelets was also inhibited by the two polyphenols as was neutrophil migration toward thrombin-induced supernatants of platelets. Quercetin and resveratrol restored the decreased CD39/ATPdase activity in human umbilical vein endothelial cells, in response to thrombin as demonstrated by adenosine monophosphate (AMP) and adenosine increases in endothelial culture supernatants. Both polyphenols inhibited thrombin-induced MAPK, JNK and focal adhesion kinase activities in endothelial cells. CONCLUSION: Quercetin and resveratrol interfere with the proinflammatory signaling of thrombin resulting in the inhibition of adenosine nucleotide secretion from activated platelets and decreased neutrophil function. Moreover, the polyphenols protect endothelial adenosine nucleotide metabolism when downregulated by thrombin. These observations may explain cardioprotective effects of grape products. PMID- 15342216 TI - In vitro evaluation of blood coagulation activation and microthrombus formation by a microchannel array flow analyzer. AB - An in vitro assay system for evaluating microthrombus formation in whole blood has not yet been established. In this study, we characterized the relationship between coagulation activation and capillary occlusion in an in vitro assay system using a microchannel array flow analyzer (MC-FAN), which can evaluate microthrombus formation in whole blood as a function of blood flow rate. The flow rate of lipopolysaccharide (LPS)-treated whole blood decreased in a dose- and time-dependent manner. Addition of LPS-pretreated mononuclear cells (PBMCs) to whole blood also decreased the flow rate. Treatment with an anti-tissue factor (TF) antibody blocked the LPS-induced decrease in blood flow rate, suggesting that TF expressed on PBMCs is responsible for decreased blood flow in the microchannels. Known anticoagulants and anti-platelet agents also inhibited the LPS-induced decrease in blood flow rate. Thrombin-antithrombin complexes were increased in LPS-stimulated whole blood, and decreased by anticoagulants but not by anti-platelet agents. These findings suggest that occlusion in microchannels occurs mainly by formation of microthrombi composed of fibrin nets and platelet aggregates, which result from TF expression on activated PBMCs. In addition, the MC-FAN was found to be useful for in vitro evaluation of antithrombotic agents in the capillary. PMID- 15342217 TI - Generation and characterization of a novel single-chain antibody fragment specific against human fibrin clots from phage display antibody library. AB - A novel single-chain fragment variable (scFv) antibody was developed directed against human fibrin clots by using the human single fold scFv libraries I+J (Tomlinson I+J). Three positively binding scFvs were evaluated by scFv-phage enzyme-linked immunosorbent assay (ELISA) and DNA sequencing. Then the positive scFv was expressed in soluble form in Escherichia coli HB2151 and purified by immobilized metal affinity chromatography (IMAC) with a yield of about 1 mg/l, the expression of soluble scFv was verified by Western blot analysis. The purified scFv could specifically recognize human fibrin clots and indicate no binding ability with human fibrinogen shown by ELISA. Furthermore, we will amplify the gene of the positive scFv by polymerase chain reaction (PCR) for future study of its role in diagnosis and therapy of thrombus-correlated diseases. PMID- 15342218 TI - An onion variety has natural antithrombotic effect as assessed by thrombosis/thrombolysis models in rodents. AB - INTRODUCTION: Prevention of arterial thrombotic diseases has a high priority in developed countries. As inappropriate diet has been shown to be an important risk factor for thrombotic events, regular antithrombotic diet may offer a convenient and effective way of prevention. The aim of the present study was to test onion extracts for antithrombotic effect and to identify the effective varieties in Allium cepa. MATERIALS AND METHODS: A shear-induced platelet function test (haemostatometry) was used to screen for antithrombotic potential. Onion extracts showing significant antithrombotic activity in vitro were further assessed in vivo by using a laser-induced thrombosis test in mice. RESULTS AND CONCLUSIONS: An onion variety, Toyohira, showed significant antithrombotic activity both in vitro and in vivo. Toyohira showed thrombolytic activity in addition to the antiplatelet effect. Superkitamomiji, 2935A, and K83211 showed only thrombolytic activity. Quercetin, the richest flavonoid in onion, was measured, but no correlation was found between quercetin content and antithrombotic activity. It is concluded that onion A. cepa can be classified into varieties with or without antithrombotic and thrombolytic effects. This should be taken into account in future population studies on the antithrombotic effects of vegetables. PMID- 15342219 TI - S-cone, L + M-cone, and pattern, electroretinograms in ocular hypertension and glaucoma. AB - Silent substitution and selective adaptation techniques were used to obtain full field S-cone and L + M-cone electroretinograms from 18 patients with ocular hypertension (OHT), 9 with normotensive glaucoma (NTG), 18 with early primary open angle glaucoma (POAG) and 19 normal controls. Pattern electroretinograms were also recorded, using a reduced check size to increase the contribution of retinal ganglion cells. In the OHT and POAG groups, statistically significant reductions (P = 0.05-0.001) were observed in the amplitudes, most notably in the late negative waves of all three types of ERG compared to the controls. These are thought to reflect ganglion cell activity. The results imply a diffusely distributed loss of activity (20-35%) affecting many retinal pathways to a similar extent in OHT and early POAG, with an additional amount (<5%) in POAG corresponding approximately to the loss associated with local field defects. The electrophysiology indicated that virtually all cases of untreated OHT have greater retinal dysfunction than the least affected cases of POAG. The NTG group showed a different pattern of loss in that the PERG was markedly affected but the S-cone ERG was not significantly reduced. PMID- 15342220 TI - Small foveal targets for studies of accommodation and the Stiles-Crawford effect. AB - The properties of small monochromatic targets as accommodative stimuli are not well understood. We used a dynamic optometer to record accommodation responses to monochromatic disc targets (1.0-27.3 min arc) and to a Maltese cross. Accommodation responded adequately to points as small as 13.6 min arc. The response to these small targets is relevant to the question of whether the Stiles Crawford (SC) effect could provide a stimulus to accommodation. Previous studies have used pupil apodizing filters to neutralise the natural SC function and so determine how visual performance or accommodation is influenced by the SC effect. However, these filters cannot correct for known inhomogeneities in the SC function across the retina for extended targets. Therefore, we calculated the SC function inhomogeneities across the retinal image of a smaller 13.6-min arc target. Unfortunately, even this small target is too large to permit a homogenous SC function across its extent. Alternatives to the apodizing filter approach are discussed. PMID- 15342221 TI - Contour grouping: closure effects are explained by good continuation and proximity. AB - Previous experimental studies have provided evidence that closed contours are easier to detect than open contours in random-element displays, and previous theoretical studies have shown that these effects might be explained by an active neural mechanism (e.g., a "reverberating neural circuit") sensitive to closure. To test this hypothesis, detection thresholds were measured in five experiments designed to control for the effects of uncertainty, eccentricity, and element density. In four of the experiments, we found that closed contours were no easier to detect than open contours, and in the remaining experiment the effects were consistent with the predictions of probability summation. Thus, we could find no evidence for an active neural mechanism that enhances detectability of closed contours more than open contours, although some form of closure mechanism may play a significant role in image interpretation. PMID- 15342222 TI - A new cue to figure-ground coding: top-bottom polarity. AB - We present evidence for a new figure-ground cue: top-bottom polarity. In an explicit reporting task, participants were more likely to interpret stimuli with a wide base and a narrow top as a figure. A similar advantage for wide-based stimuli also occurred in a visual short-term memory task, where the stimuli had ambiguous figure-ground relations. Further support comes from a figural search task. Figural search is a discrimination task in which participants are set to search for a symmetric target in a display with ambiguous figure-ground organization. We show that figural search was easier when stimuli with a top bottom polarity were placed in an orientation where they had a wide base and a narrow top, relative to when this orientation was inverted. This polarity effect was present when participants were set to use color to parse figure from ground, and it was magnified when the participants did not have any foreknowledge of the color of the symmetric target. Taken together the results suggest that top-bottom polarity influences figure-ground assignment, with wide base stimuli being preferred as a figure. In addition, the figural search task can serve as a useful procedure to examine figure-ground assignment. PMID- 15342223 TI - Retinal nerve fiber layer thinning in Parkinson disease. AB - Retinal dopamine loss in Parkinson disease (PD) is reflected by visual neurophysiological dysfunction. We measured the thickness of the circumpapillary retinal nerve fiber layer (RNFL) in PD patients using optical coherence tomography. The thickness in the inferior quadrant of PD patients (147 +/- 20 microns) was significantly thinner than that of controls (173 +/- 12 microns; p=0.002), while the inferotemporal area was the thinnest (146 +/- 24 vs. 191 +/- 21 microns; p=0.0003). The results show significant loss of RNFL thickness in PD at specific sites. PMID- 15342224 TI - Models of the mechanism underlying perceived location of a perisaccadic flash. AB - A variety of experiments have shown that subjects tend to perceive a target flash as mislocalized when the flash is presented just before, during or shortly after the occurrence of a saccade. The characteristics of this mislocalization suggest that it arises from an anticipatory, slow extraretinal signal, i.e., the signal starts to change before a saccade and continues to change during and after the saccade. However, a target flash creates a visual signal that can persist for as long as 300 ms. Interaction of this visual persistence with the extraretinal signal could have a significant influence on the perceived location of the target flash, and thus on features of the extraretinal signal as inferred from the perceived location. In this study, several different types of models were used to explore how retinal signal persistence together with an extraretinal signal might affect perception. According to these models, the anticipatory, slow extraretinal signal may be an artifact of using a target flash, and the actual extraretinal signal may begin to change only after saccade onset and relatively quickly. PMID- 15342225 TI - Curvature population coding for complex shapes in human vision. AB - In the primate visual system relatively complex patterns such as curved shapes are first represented at intermediate levels of the ventral pathway. Furthermore, there is now evidence for the existence of curvature population coding in primate V4. We sought to determine whether similar encoding occurs in the human visual system by using a context-dependent lateral masking paradigm. In this paradigm a central closed contour comprising the test pattern is masked by surrounding larger or smaller patterns with various configurations. Results indicate that test thresholds are not affected by a circular control mask, and that elevations are greatest when curvature extrema of the mask are aligned with those of the target. These lateral interactions extend over greater than 1 degrees and are tuned for target shape. Masking increases with the number of local curvature extrema aligned with the target. Finally, masking persists when target and mask have orthogonal local orientations and increases with mask amplitude. These findings are incompatible with local orientation-selective interactions (V1 mediated) but are consistent with the existence of population codes based on curvature maxima at intermediate levels of processing (presumably V4) in human vision. The paradigm we introduce provides a new tool for evaluating the representation of complex percepts. PMID- 15342226 TI - Convergence reduces ocular counterroll (OCR) during static roll-tilt. AB - When humans are roll-tilted around the naso-occipital axis, both eyes roll or tort in the opposite direction to roll-tilt, a phenomenon known as ocular counterroll (OCR). While the magnitude of OCR is primarily determined by vestibular, somatosensory, and proprioceptive input, direction of gaze also plays a major role. The aim of this study was to measure the interaction between some of these factors in the control of OCR. Videooculography was used to measure 3D eye position during maintained whole body (en bloc) static roll-tilt in darkness, while subjects fixated first on a distant (at 130 cm) and then a near (at 30 cm) head-fixed target aligned with the subject's midline. We found that while converging on the near target, human subjects displayed a significant reduction in OCR for both directions of roll-tilt--i.e. the interaction between OCR and vergence was not simple addition or subtraction of torsion induced by vergence with torsion induced by roll-tilt. To remove the possibility that the OCR reduction may be associated with the changed horizontal position of the eye in the orbit during symmetric convergence, we ran an experiment using asymmetric convergence in which the distant and near targets were aligned directly in front of one eye. We found the magnitude of OCR in this asymmetric convergence case was also reduced for near viewing by about the same amount as in the symmetric vergence condition, confirming that the convergence command rather than horizontal position of the eye underlies the OCR reduction, since there was no horizontal movement of the aligned eye in the orbit between fixation on the distant and near targets. Increasing vergence from 130 to 30 cm reduced OCR gain by around 35% on average. That reduction was equal in both eyes and occurred in both the symmetric and asymmetric convergence conditions. These results demonstrate the important role vergence plays in determining ocular counterroll during roll-tilt and may support the contention that vergence acts to reduce the conflict facing a stereopsis-generating mechanism. PMID- 15342227 TI - Texture segregation in traumatic brain injury--a VEP study. AB - Visual evoked potentials (VEPs) were recorded to textures segregated by gradients in orientation or motion. Recordings were obtained in traumatic brain-injured (TBI) subjects and in normal controls. We analyzed both the low-level VEPs (llVEPs) evoked by homogenous stimuli, as well as the components associated with texture segregation (tsVEP) obtained through an appropriate linear combination. Our results suggest that the tsVEP, presumably higher up in the visual processing chain than the llVEP, is sensitive to TBI and can reveal further information as to the nature of possible information processing deficits after TBI. It could also help quantify cortical damage that is not revealed with more standard clinical tools. PMID- 15342228 TI - Modeling color percepts of dichromats. AB - Protanopes and deuteranopes, despite lacking a chromatic dimension at the receptor level, use the color terms "red" and "green", together with "blue" and "yellow", to describe their color percepts. Color vision models proposed so far fail to account for these findings in dichromats. We confirmed, by the method of hue scaling, the consistent use of these color terms, as well as their dependence on intensity, in subjects shown to have only a single X-chromosomal opsin gene each. We present a model for the processing of photoreceptor signals which, under physiologically plausible assumptions, achieves a trichromat-like representation of dichromatic receptor signals. Key feature of the dichromat model is the processing of the photoreceptor signals in parallel channels with different gains and nonlinearities. In this way, the two-dimensional receptor signals are represented on a manifold in a higher-dimensional space, supporting categorization for efficient image segmentation. Introducing a third cone opsin yields a model that explains normal, trichromat hue scaling. PMID- 15342229 TI - Linking motion-induced blindness to perceptual filling-in. AB - "Motion-induced blindness" and "perceptual filing-in" are two phenomena in which perceptually salient stimuli repeatedly disappear and reappear after prolonged viewing. Despite the many similarities between MIB and PFI, two differences suggest that they could be unrelated phenomena: (1) An area surrounded by background stimuli can be perceived to disappear completely in PFI but not in MIB and (2) high contrast stimuli are perceived to disappear less easily in PFI but, remarkably enough, more easily in MIB. In this article we show that the apparent differences between MIB and PFI disappear when eccentricity, contrast, and perceptual grouping are taken into account and that both are most likely caused by the same underlying mechanism. PMID- 15342230 TI - Structure of the ligand-blocked periplasmic entrance of the bacterial multidrug efflux protein TolC. AB - The trimeric TolC protein of Escherichia coli comprises an outer membrane beta barrel and a contiguous alpha-helical barrel projecting across the periplasm. This provides a single 140 A long pore for multidrug efflux and protein export. We have previously reported that trivalent cations such as hexammine cobalt can severely inhibit the conductivity of the TolC pore reconstituted in planar lipid bilayers. Here, isothermal calorimetry shows that Co(NH(3))(6)(3+) binds to TolC with an affinity of 20 nM. The crystal structure of the TolC-Co(NH(3))(6)(3+) complex was determined to 2.75 A resolution, and showed no significant difference in the protein when compared with unliganded TolC. An electron density difference map revealed that a single ligand molecule binds at the centre of the periplasmic entrance, the sole constriction of TolC. The octahedral symmetry of the ligand and the three-fold rotational symmetry of the TolC entrance determine a binding site in which the ligand forms hydrogen bonds with the Asp(374) residue of each monomer. When Asp(374) was substituted by alanine, high affinity ligand binding was abolished and inhibition of TolC pore conductivity in lipid bilayers was alleviated. Comparable effects followed independent substitution of the neighbouring Asp(371), indicating that this aspartate ring also contributes to the high affinity ligand binding site. As the electronegative entrance is widely conserved in the TolC family, it may be a useful target for the development of inhibitors against multidrug resistant pathogenic bacteria. PMID- 15342231 TI - Reversible unfolding of beta-sheets in membranes: a calorimetric study. AB - The hexapeptide acetyl-Trp-Leu(5) (AcWL(5)) has the remarkable ability to assemble reversibly and spontaneously into beta-sheets on lipid membranes as a result of monomer partitioning followed by cooperative assembly. This system provides a unique opportunity to study the thermodynamics of protein folding in membranes, which we have done using isothermal titration calorimetry (ITC) and differential scanning calorimetry (DSC). The results, which may represent the first example of reversible thermal unfolding of peptides in membranes, help to define the contribution of hydrogen bonding to the extreme thermal stability of membrane proteins. ITC revealed that the enthalpy change for partitioning of monomeric, unstructured AcWL(5) from water into membranes was zero within experimental error over the temperature range of 5 degrees C to 75 degrees C. DSC showed that the beta-sheet aggregates underwent a reversible, endothermic, and very asymmetric thermal transition with a concentration-dependent transition temperature (T(m)) in the range of 60 degrees C to 80 degrees C. A numerical model of nucleation and growth-dependent assembly of oligomeric beta-sheets, proposed earlier to describe beta-sheet formation in membranes, recreated remarkably well the unusual shape and concentration-dependence of the transition peaks. The enthalpy for thermal unfolding of AcWL(5) beta-sheets in the membrane was found to be about 8(+/-1)kcal mol(-1), or about 1.3(+/-0.2)kcal mol(-1) per residue. PMID- 15342232 TI - On the role of the starved codon and the takeoff site in ribosome bypassing in Escherichia coli. AB - Translating ribosomes can skip over stretches of messenger RNA and resume protein chain elongation after a "bypassed" region. We have previously shown that limitation for isoleucyl-tRNA can initiate a ribosome bypass when an AUA codon is in the ribosomal A-site. We have now generalized this effect to other "hungry" codons calling for four different limiting aminoacyl-tRNA species, suggesting that a pause at any A-site will have this effect. We have assessed bypassing in a large family of reporters with nearly every different triplet in the "takeoff site", i.e. the P-site on the 5' side of the hungry codon, and an identical "landing site" codon 16 nucleotides downstream. The different takeoff sites vary over a factor of 50 in bypassing proficiency. At least part of this variation appears to reflect stability of the codon Colon, two colons anticodon interaction at the takeoff site, as indicated by the following: (a) the bypassing proficiency of different tRNAs shows a rough correlation with the frequency of A Colon, two colons U as opposed to G Colon, two colons C pairs in the codon Colon, two colons anticodon association; (b) specific tRNAs bypass more frequently from codons ending in U than from their synonym ending in C; (c) an arginine tRNA with Inosine in the wobble position which reads CGU, CGC, and CGA bypasses much more frequently from the last codon than the first two synonyms. PMID- 15342233 TI - Importance of transient structures during post-transcriptional refolding of the pre-23S rRNA and ribosomal large subunit assembly. AB - An important step of ribosome assembly is the folding of the rRNA into a functional structure. Despite knowledge of the folded state of rRNA in the ribosomal subunits, there is very little information on the rRNA folding pathway. We are interested in understanding how the functional structure of rRNA is formed and whether the rRNA folding intermediates have a role in ribosome assembly. To this end, transient secondary structures around both ends of pre-23S rRNA were analyzed by a chemical probing approach, using pre-23S rRNA transcripts. Metastable hairpin loop structures were found at both ends of 23S rRNA. The functional importance of the transient structures around the ends of 23S rRNA was tested by mutations that alter only the transient structure. The effect of mutations on 23S rRNA folding was tested in vitro and in vivo. It was found that both stabilization and destabilization of the transient structure around the 5' end of 23S rRNA inhibits post-transcriptional refolding in vitro and ribosome formation in vivo. The data suggest that the transient structure of rRNA has a function during 23S rRNA folding and thereby in ribosome assembly. PMID- 15342234 TI - Human UP1 as a model for understanding purine recognition in the family of proteins containing the RNA recognition motif (RRM). AB - Heterogeneous ribonucleoprotein A1 (hnRNP A1) is a prototype for the family of eukaryotic RNA processing proteins containing the common RNA recognition motif (RRM). The region consisting of residues 1-195 of hnRNP A1 is referred to as UP1. This region has two RRMs and has a high affinity for both single-stranded RNA and the human telomeric repeat sequence d(TTAGGG)(n). We have used UP1's novel DNA binding to investigate how RRMs bind nucleic acid bases through their highly conserved RNP consensus sequences. Nine complexes of UP1 bound to modified telomeric repeats were investigated using equilibrium fluorescence binding and X ray crystallography. In two of the complexes, alteration of a guanine to either 2 aminopurine or nebularine resulted in an increase in K(d) from 88nM to 209nM and 316nM, respectively. The loss of these orienting interactions between UP1 and the substituted base allows it to flip between syn and anti conformations. Substitution of the same base with 7-deaza-guanine preserves the O6/N1 contacts but still increases the K(d) to 296nM and suggests that it is not simply the loss of affinity that gives rise to the base mobility, but also the stereochemistry of the specific contact to O6. Although these studies provide details of UP1 interactions to nucleic acids, three general observations about RRMs are also evident: (1) as suggested by informatic studies, main-chain to base hydrogen bonding makes up an important aspect of ligand recognition (2) steric clashes generated by modification of a hydrogen bond donor-acceptor pair to a donor-donor pair are poorly tolerated and (3) a conserved lysine position proximal to RNP-2 (K(106)-IFVGGI) orients the purine to allow stereochemical discrimination between adenine and guanine based on the 6-position. This single interaction is well conserved in known RRM structures and appears to be a broad indicator for purine preference in the larger family of RRM proteins. PMID- 15342235 TI - The archaeal sRNA binding protein L7Ae has a 3D structure very similar to that of its eukaryal counterpart while having a broader RNA-binding specificity. AB - The ribosomal L7Ae protein of archaea has the peculiarity to be a component of the C/D and H/ACA snRNPs, that guide rRNA post-transcriptional modifications. Its yeast (Snu13p) and human (15.5kDa protein) homologs are only found in C/D snoRNPs and the (U4/U6, U5) spliceosomal tri-snRNP. By using a large variety of RNAs, we compared the RNA-binding specificities of the recombinant Pyrococcus abyssi L7Ae and Saccharomyces cerevisiae Snu13 proteins. Unlike Snu13p, protein L7Ae binds terminal loops closed by two A:G and G:A pairs and canonical K-turn structures with similar efficiencies, provided that the terminal loop contains at least 5nt. In contrast to Snu13p, binding of protein L7Ae to canonical K-turn structures is not dependent on the identity of the residue at position 2 in the bulge. The peculiar KT-15 motif of P. abyssi 23S rRNA, that is recognized by L7Ae, does not associate with Snu13p. To get more information on the P. abyssi L7Ae protein, we solved its X-ray structure at 1.9A resolution. In spite of their sequence divergence, the free P. abyssi and bound H. marismortui proteins were found to have highly similar structures. Only a limited number of side-chain conformational changes occur at the protein-RNA interface upon RNA binding. In particular, one ion pair that is formed by residues Glu43 and Lys46 in the free protein is disrupted in the ribosomal 50S subunit, so that, residue Glu43 can interact with the RNA residue G264. The Glu43-Lys46 ion pair of protein L7Ae belongs to a complex network of ion pairs that may participate to protein thermostability. PMID- 15342236 TI - Stacked-unstacked equilibrium at the nick site of DNA. AB - Stability of duplex DNA with respect to separation of complementary strands is crucial for DNA executing its major functions in the cell and it also plays a central role in major biotechnology applications of DNA: DNA sequencing, polymerase chain reaction, and DNA microarrays. Two types of interaction are well known to contribute to DNA stability: stacking between adjacent base-pairs and pairing between complementary bases. However, their contribution into the duplex stability is yet to be determined. Now we fill this fundamental gap in our knowledge of the DNA double helix. We have prepared a series of 32, 300 bp-long DNA fragments with solitary nicks in the same position differing only in base pairs flanking the nick. Electrophoretic mobility of these fragments in the gel has been studied. Assuming the equilibrium between stacked and unstacked conformations at the nick site, all 32 stacking free energy parameters have been obtained. Only ten of them are essential and they govern the stacking interactions between adjacent base-pairs in intact DNA double helix. A full set of DNA stacking parameters has been determined for the first time. From these data and from a well-known dependence of DNA melting temperature on G.C content, the contribution of base-pairing into duplex stability has been estimated. The obtained energy parameters of the DNA double helix are of paramount importance for understanding sequence-dependent DNA flexibility and for numerous biotechnology applications. PMID- 15342237 TI - Trypanosoma cruzi contains a single detectable uracil-DNA glycosylase and repairs uracil exclusively via short patch base excision repair. AB - Enzymes involved in genomic maintenance of human parasites are attractive targets for parasite-specific drugs. The parasitic protozoan Trypanosoma cruzi contains at least two enzymes involved in the protection against potentially mutagenic uracil, a deoxyuridine triphosphate nucleotidohydrolase (dUTPase) and a uracil DNA glycosylase belonging to the highly conserved UNG-family. Uracil-DNA glycosylase activities excise uracil from DNA and initiate a multistep base excision repair (BER) pathway to restore the correct nucleotide sequence. Here we report the biochemical characterisation of T.cruzi UNG (TcUNG) and its contribution to the total uracil repair activity in T.cruzi. TcUNG is shown to be the major uracil-DNA glycosylase in T.cruzi. The purified recombinant TcUNG exhibits substrate preference for removal of uracil in the order ssU>U:G>U:A, and has no associated thymine-DNA glycosylase activity. T.cruzi apparently repairs U:G DNA substrate exclusively via short-patch BER, but the DNA polymerase involved surprisingly displays a vertebrate POLdelta-like pattern of inhibition. Back-up UDG activities such as SMUG, TDG and MBD4 were not found, underlying the importance of the TcUNG enzyme in protection against uracil in DNA and as a potential target for drug therapy. PMID- 15342238 TI - Regulation of DNA binding of p53 by its C-terminal domain. AB - The tumor suppressor p53 is a tetrameric multi-domain transcription factor. Its C terminal domain is thought to regulate the binding of its core domain to specific recognition sequences in promoters. The mechanism of regulation by the C-terminal domain and the role of its post-translational modification are controversial. We have examined the binding of DNA in solution to a series of unmodified p53 constructs that lack various domains. The specific DNA sequences bind tightly to the core domain, irrespective of whether or not the C-terminal domain is part of the construct. Unmodified p53 is accordingly an active DNA binding protein. Non specific DNA sequences do not inhibit directly the binding of the specific sequences to the core but bind to the C terminus and inhibit p53 via that binding mode. Using NMR, we identified the residues of the C terminus that interact with the non-specific DNA. They include residues that are known to be modified post translationally. Our data provide direct support for the regulatory role of the C terminus in the activity of p53 and show that p53 containing the unmodified C terminus actively binds to short double-stranded DNA. PMID- 15342239 TI - The NMR solution structure of a mutant of the Max b/HLH/LZ free of DNA: insights into the specific and reversible DNA binding mechanism of dimeric transcription factors. AB - Basic region-helix1-loop-helix2-leucine zipper (b/H(1)LH(2)/LZ) transcription factors bind specific DNA sequence in their target gene promoters as dimers. Max, a b/H(1)LH(2)/LZ transcription factor, is the obligate heterodimeric partner of the related b/H(1)LH(2)/LZ proteins of the Myc and Mad families. These heterodimers specifically bind E-box DNA sequence (CACGTG) to activate (e.g. c Myc/Max) and repress (e.g. Mad1/Max) transcription. Max can also homodimerize and bind E-box sequences in c-Myc target gene promoters. While the X-ray structure of the Max b/H(1)LH(2)/LZ/DNA complex and that of others have been reported, the precise sequence of events leading to the reversible and specific binding of these important transcription factors is still largely unknown. In order to provide insights into the DNA binding mechanism, we have solved the NMR solution structure of a covalently homodimerized version of a Max b/H(1)LH(2)/LZ protein with two stabilizing mutations in the LZ, and characterized its backbone dynamics from (15)N spin-relaxation measurements in the absence of DNA. Apart from minor differences in the pitch of the LZ, possibly resulting from the mutations in the construct, we observe that the packing of the helices in the H(1)LH(2) domain is almost identical to that of the two crystal structures, indicating that no important conformational change in these helices occurs upon DNA binding. Conversely to the crystal structures of the DNA complexes, the first 14 residues of the basic region are found to be mostly unfolded while the loop is observed to be flexible. This indicates that these domains undergo conformational changes upon DNA binding. On the other hand, we find the last four residues of the basic region form a persistent helical turn contiguous to H(1). In addition, we provide evidence of the existence of internal motions in the backbone of H(1) that are of larger amplitude and longer time-scale (nanoseconds) than the ones in the H(2) and LZ domain. Most interestingly, we note that conformers in the ensemble of calculated structures have highly conserved basic residues (located in the persistent helical turn of the basic region and in the loop) known to be important for specific binding in a conformation that matches that of the DNA bound state. These partially prefolded conformers can directly fit into the major groove of DNA and as such are proposed to lie on the pathway leading to the reversible and specific DNA binding. In these conformers, the conserved basic side-chains form a cluster that elevates the local electrostatic potential and could provide the necessary driving force for the generation of the internal motions localized in the H(1) and therefore link structural determinants with the DNA binding function. Overall, our results suggests that the Max homodimeric b/H(1)LH(2)/LZ can rapidly and preferentially bind DNA sequence through transient and partially prefolded states and subsequently, adopt the fully helical bound state in a DNA-assisted mechanism or induced-fit. PMID- 15342240 TI - Expressed gene clusters associated with cellular sensitivity and resistance towards anti-viral and anti-proliferative actions of interferon. AB - Interferons (IFN) are multi-functional proteins that induce a large number of genes which mediate many biological processes including host defense, cell growth control, signaling, and metabolism. Bioinformatics analysis of the 3' untranslated regions of IFN-stimulated genes (ISGs) showed that the AU-rich elements (ARE) exist in approximately 10% of the mRNA induced by IFN. The human epithelial cell lines, WISH and 293, and the human B cell lines, Daudi and RPMI 1788, were assessed for their response to type-I IFN. Due to their differential response to the anti-viral and anti-proliferative action of IFN-alpha, they were used as cellular models for genome wide ARE-gene expression. The anti-viral and anti-proliferative actions of IFN-alpha were substantially more potent against WISH and Daudi cells than 293 and RPMI 1788 cells, respectively. These results correlated with the Stat1-driven gene expression as assessed by monitoring the expression of Stat1-mediated IFN-inducible 6-16 mRNA. Interferons were able to induce a significant proportion of common and distinct ARE-genes, but the patterns of expression were different and dependent on the type of the cell, type of IFN, and status of the cellular sensitivity to IFN. Clustering algorithms generated two informative expressed gene clusters that were selectively associated with cellular sensitivity and resistance to the anti-viral and anti proliferative action of IFN. Use of rationally designed microarray experiments in IFN biology yielded informative clusters that may provide candidate genes for diagnostic or for evaluation of therapeutic possibilities. PMID- 15342241 TI - The crystal structure of mouse phosphoglucose isomerase at 1.6A resolution and its complex with glucose 6-phosphate reveals the catalytic mechanism of sugar ring opening. AB - Phosphoglucose isomerase (PGI) is an enzyme of glycolysis that interconverts glucose 6-phosphate (G6P) and fructose 6-phosphate (F6P) but, outside the cell, is a multifunctional cytokine. High-resolution crystal structures of the enzyme from mouse have been determined in native form and in complex with the inhibitor erythrose 4-phosphate, and with the substrate glucose 6-phosphate. In the substrate-bound structure, the glucose sugar is observed in both straight-chain and ring forms. This structure supports a specific role for Lys518 in enzyme catalyzed ring opening and we present a "push-pull" mechanism in which His388 breaks the O5-C1 bond by donating a proton to the ring oxygen atom and, simultaneously, Lys518 abstracts a proton from the C1 hydroxyl group. The reverse occurs in ring closure. The transition from ring form to straight-chain substrate is achieved through rotation of the C3-C4 bond, which brings the C1-C2 region into close proximity to Glu357, the base catalyst for the isomerization step. The structure with G6P also explains the specificity of PGI for glucose 6-phosphate over mannose 6-isomerase (M6P). To isomerize M6P to F6P requires a rotation of its C2-C3 bond but in PGI this is sterically blocked by Gln511. PMID- 15342242 TI - Structure and function of a regulated archaeal triosephosphate isomerase adapted to high temperature. AB - Triosephophate isomerase (TIM) is a dimeric enzyme in eucarya, bacteria and mesophilic archaea. In hyperthermophilic archaea, however, TIM exists as a tetramer composed of monomers that are about 10% shorter than other eucaryal and bacterial TIM monomers. We report here the crystal structure of TIM from Thermoproteus tenax, a hyperthermophilic archaeon that has an optimum growth temperature of 86 degrees C. The structure was determined from both a hexagonal and an orthorhombic crystal form to resolutions of 2.5A and 2.3A, and refined to R-factors of 19.7% and 21.5%, respectively. In both crystal forms, T.tenax TIM exists as a tetramer of the familiar (betaalpha)(8)-barrel. In solution, however, and unlike other hyperthermophilic TIMs, the T.tenax enzyme exhibits an equilibrium between inactive dimers and active tetramers, which is shifted to the tetramer state through a specific interaction with glycerol-1-phosphate dehydrogenase of T.tenax. This observation is interpreted in physiological terms as a need to reduce the build-up of thermolabile metabolic intermediates that would be susceptible to destruction by heat. A detailed structural comparison with TIMs from organisms with growth optima ranging from 15 degrees C to 100 degrees C emphasizes the importance in hyperthermophilic proteins of the specific location of ionic interactions for thermal stability rather than their numbers, and shows a clear correlation between the reduction of heat-labile, surface exposed Asn and Gln residues with thermoadaptation. The comparison confirms the increase in charged surface-exposed residues at the expense of polar residues. PMID- 15342243 TI - Atomic force microscopy reveals defects within mica supported lipid bilayers induced by the amyloidogenic human amylin peptide. AB - To date, over 20 peptides or proteins have been identified that can form amyloid fibrils in the body and are thought to cause disease. The mechanism by which amyloid peptides cause the cytotoxicity observed and disease is not understood. However, one of the major hypotheses is that amyloid peptides cause membrane perturbation. Hence, we have studied the interaction between lipid bilayers and the 37 amino acid residue polypeptide amylin, which is the primary constituent of the pancreatic amyloid associated with type 2 diabetes. Using a dye release assay we confirmed that the amyloidogenic human amylin peptide causes membrane disruption; however, time-lapse atomic force microscopy revealed that this did not occur by the formation of defined pores. On the contrary, the peptide induced the formation of small defects spreading over the lipid surface. We also found that rat amylin, which has 84% identity with human amylin but cannot form amyloid fibrils, could also induce similar lesions to supported lipid bilayers. The effect, however, for rat amylin but not human amylin, was inhibited under high ionic conditions. These data provide an alternative theory to pore formation, and how amyloid peptides may cause membrane disruption and possibly cytotoxicity. PMID- 15342244 TI - Crystal structure of an activation intermediate of cathepsin E. AB - Cathepsin E is an intracellular, non-lysosomal aspartic protease expressed in a variety of cells and tissues. The protease has proposed physiological roles in antigen presentation by the MHC class II system, in the biogenesis of the vasoconstrictor peptide endothelin, and in neurodegeneration associated with brain ischemia and aging. Cathepsin E is the only A1 aspartic protease that exists as a homodimer with a disulfide bridge linking the two monomers. Like many other aspartic proteases, it is synthesized as a zymogen which is catalytically inactive towards its natural substrates at neutral pH and which auto-activates in an acidic environment. Here we report the crystal structure of an activation intermediate of human cathepsin E at 2.35A resolution. The overall structure follows the general fold of aspartic proteases of the A1 family, and the intermediate shares many features with the intermediate 2 on the proposed activation pathway of aspartic proteases like pepsin C and cathepsin D. The pro sequence is cleaved from the protease and remains stably associated with the mature enzyme by forming the outermost sixth strand of the interdomain beta sheet. However, different from these other aspartic proteases the pro-sequence of cathepsin E remains intact after cleavage from the mature enzyme. In addition, the active site of cathepsin E in the crystal is occupied by N-terminal amino acid residues of the mature protease in the non-primed binding site and by an artificial N-terminal extension of the pro-sequence from a neighboring molecule in the primed site. The crystal structure of the cathepsin E/pro-sequence complex, therefore, provides further insight into the activation mechanism of aspartic proteases. PMID- 15342245 TI - Development of a human light chain variable domain (V(L)) intracellular antibody specific for the amino terminus of huntingtin via yeast surface display. AB - Intracellular antibodies (intrabodies) provide an attractive means for manipulating intracellular protein function, both for research and potentially for therapy. A challenge in the isolation of effective intrabodies is the ability to find molecules that exhibit sufficient binding affinity and stability when expressed in the reducing environment of the cytoplasm. Here, we have used yeast surface display of proteins to isolate novel scFv clones against huntingtin from a non-immune human antibody library. We then applied yeast surface display to affinity mature this scFv pool and analyze the location of the binding site of the mutant with the highest affinity. Interestingly, the paratope was mapped exclusively to the variable light chain domain of the scFv. A single domain antibody was constructed consisting solely of this variable light chain domain, and was found to retain full binding activity to huntingtin. Cytoplasmic expression levels in yeast of the single domain were at least fivefold higher than the scFv. The ability of the single-domain intrabody to inhibit huntingtin aggregation, which has been implicated in the pathogenesis of Huntington's disease (HD), was confirmed in a cell-free in vitro assay as well as in a mammalian cell culture model of HD. Significantly, a single-domain intrabody that is functionally expressable in the cytoplasm was derived from a non-functional scFv by performing affinity maturation and binding site analysis on the yeast cell surface, despite the differences between the cytoplasmic and extracellular environment. This approach may find application in the development of intrabodies to a wide variety of intracellular targets. PMID- 15342246 TI - A comparison of the dynamic behavior of monomeric and dimeric insulin shows structural rearrangements in the active monomer. AB - Molecular dynamics (MD) simulations (5-10ns in length) and normal mode analyses were performed for the monomer and dimer of native porcine insulin in aqueous solution; both starting structures were obtained from an insulin hexamer. Several simulations were done to confirm that the results obtained are meaningful. The insulin dimer is very stable during the simulation and remains very close to the starting X-ray structure; the RMS fluctuations calculated from the MD simulation agree with the experimental B-factors. Correlated motions were found within each of the two monomers; they can be explained by persistent non-bonded interactions and disulfide bridges. The correlated motions between residues B24 and B26 of the two monomers are due to non-bonded interactions between the side-chains and backbone atoms. For the isolated monomer in solution, the A chain and the helix of the B chain are found to be stable during 5ns and 10ns MD simulations. However, the N-terminal and the C-terminal parts of the B chain are very flexible. The C-terminal part of the B chain moves away from the X-ray conformation after 0.5-2.5ns and exposes the N-terminal residues of the A chain that are thought to be important for the binding of insulin to its receptor. Our results thus support the hypothesis that, when monomeric insulin is released from the hexamer (or the dimer in our study), the C-terminal end of the monomer (residues B25-B30) is rearranged to allow binding to the insulin receptor. The greater flexibility of the C-terminal part of the beta chain in the B24 (Phe- >Gly) mutant is in accord with the NMR results. The details of the backbone and side-chain motions are presented. The transition between the starting conformation and the more dynamic structure of the monomers is characterized by displacements of the backbone of Phe B25 and Tyr B26; of these, Phe B25 has been implicated in insulin activation. PMID- 15342247 TI - How small peptides block and reverse serpin polymerisation. AB - Many of the late-onset dementias, including Alzheimer's disease and the prion encephalopathies, arise from the aberrant aggregation of individual proteins. The serpin family of serine protease inhibitors provides a well-defined structural example of such pathological aggregation, as its mutant variants readily form long-chain polymers, resulting in diseases ranging from thrombosis to dementia. The intermolecular linkages result from the insertion of the reactive site loop of one serpin molecule into the middle strand (s4A) position of the A beta-sheet of another molecule. We define here the structural requirements for small peptides to competitively bind to and block the s4A position to prevent this intermolecular linkage and polymerisation. The entry and anchoring of blocking peptides is facilitated by the presence of a threonine which inserts into the site equivalent to P8 of s4A. But the critical requirement for small blocking peptides is demonstrated in crystallographic structures of the complexes formed with selected tri- and tetrapeptides. These structures indicate that the binding is primarily due to the insertion of peptide hydrophobic side-chains into the P4 and P6 sites of s4A. The findings allow the rational design of synthetic blocking peptides small enough to be suitable for mimetic design. This is demonstrated here with a tetrapeptide that preferentially blocks the polymerisation of a pathologically unstable serpin commonly present in people of European descent. PMID- 15342248 TI - Crystal structure of human ABAD/HSD10 with a bound inhibitor: implications for design of Alzheimer's disease therapeutics. AB - The enzyme 17beta-hydroxysteroid dehydrogenase type 10 (HSD10), also known as amyloid beta-peptide-binding alcohol dehydrogenase (ABAD), has been implicated in the development of Alzheimer's disease. This protein, a member of the short-chain dehydrogenase/reductase family of enzymes, has been shown to bind beta-amyloid and to participate in beta-amyloid neurotoxicity. We have determined the crystal structure of human ABAD/HSD10 complexed with NAD(+) and an inhibitory small molecule. The inhibitor occupies the substrate-binding site and forms a covalent adduct with the NAD(+) cofactor. The crystal structure provides a basis for the design of potent, highly specific ABAD/HSD10 inhibitors with potential application in the treatment of Alzheimer's disease. PMID- 15342249 TI - Structure of Mycobacterium tuberculosis FtsZ reveals unexpected, G protein-like conformational switches. AB - We report three crystal structures of the Mycobacterium tuberculosis cell division protein FtsZ, as the citrate, GDP, and GTPgammaS complexes, determined at 1.89, 2.60, and 2.08A resolution. MtbFtsZ crystallized as a tight, laterally oriented dimer distinct from the longitudinal polymer observed for alphabeta tubulin. Mutational data on Escherichia coli FtsZ suggest that this dimer interface is important for proper protofilament and "Z-ring" assembly and function. An alpha-to-beta secondary structure conformational switch at the dimer interface is spatially analogous to, and has many of the hallmarks of, the Switch I conformational changes exhibited by G-proteins upon activation. The presence of a gamma-phosphate in the FtsZ active site modulates the conformation of the "tubulin" loop T3 (spatially analogous to the G-protein Switch II); T3 switching upon gamma-phosphate ligation is directly coupled to the alpha-to-beta switch by steric overlap. The dual conformational switches observed here for the first time in an FtsZ link GTP binding and hydrolysis to FtsZ (and tubulin) lateral assembly and Z-ring contraction, and they are suggestive of an underappreciated functional analogy between FtsZ, tubulin and G-proteins. PMID- 15342250 TI - Unusual Cys-Tyr covalent bond in a large catalase. AB - Catalase-1, one of four catalase activities of Neurospora crassa, is associated with non-growing cells and accumulates in asexual spores. It is a large, tetrameric, highly efficient, and durable enzyme that is active even at molar concentrations of hydrogen peroxide. Catalase-1 is oxidized at the heme by singlet oxygen without significant effects on enzyme activity. Here we present the crystal structure of catalase-1 at 1.75A resolution. Compared to structures of other catalases of the large class, the main differences were found at the carboxy-terminal domain. The heme group is rotated 180 degrees around the alpha gamma-meso carbon axis with respect to clade 3 small catalases. There is no co ordination bond of the ferric ion at the heme distal side in catalase-1. The catalase-1 structure exhibited partial oxidation of heme b to heme d. Singlet oxygen, produced catalytically or by photosensitization, may hydroxylate C5 and C6 of pyrrole ring III with a subsequent formation of a gamma-spirolactone in C6. The modification site in catalases depends on the way dioxygen exits the protein: mainly through the central channel or the main channel in large and small catalases, respectively. The catalase-1 structure revealed an unusual covalent bond between a cysteine sulphur atom and the essential tyrosine residue of the proximal side of the active site. A peptide with the predicted theoretical mass of the two bound tryptic peptides was detected by mass spectrometry. A mechanism for the Cys-Tyr covalent bond formation is proposed. The tyrosine bound to the cysteine residue would be less prone to donate electrons to compound I to form compound II, explaining catalase-1 resistance to substrate inhibition and inactivation. An apparent constriction of the main channel at Ser198 lead us to propose a gate that opens the narrow part of the channel when there is sufficient hydrogen peroxide in the small cavity before the gate. This mechanism would explain the increase in catalytic velocity as the hydrogen peroxide concentration rises. PMID- 15342251 TI - T=1 capsid structures of Sesbania mosaic virus coat protein mutants: determinants of T=3 and T=1 capsid assembly. AB - Sesbania mosaic virus particles consist of 180 coat protein subunits of 29kDa organized on a T=3 icosahedral lattice. N-terminal deletion mutants of coat protein that lack 36 (CP-NDelta36) and 65 (CP-NDelta65) residues from the N terminus, when expressed in Escherichia coli, produced similar T=1 capsids of approximate diameter 20nm. In contrast to the wild-type particles, these contain only 60 copies of the truncated protein subunits (T=1). CP-NDelta65 lacks the "beta-annulus" believed to be responsible for the error-free assembly of T=3 particles. Though the CP-NDelta36 mutant has the beta-annulus segment, it does not form a T=3 capsid, presumably because it lacks an arginine-rich motif found close to the amino terminus. Both CP-NDelta36 and CP-NDelta65 T=1 capsids retain many key features of the T=3 quaternary structure. Calcium binding geometries at the coat protein interfaces in these two particles are also nearly identical. When the conserved aspartate residues that coordinate the calcium, D146 and D149 in the CP-NDelta65, were mutated to asparagine (CP-NDelta65-D146N-D149N), the subunits assembled into T=1 particles but failed to bind calcium ions. The structure of this mutant revealed particles that were slightly expanded. The analysis of the structures of these mutant capsids suggests that although calcium binding contributes substantially to the stability of T=1 particles, it is not mandatory for their assembly. In contrast, the presence of a large fraction of the amino-terminal arm including sequences that precede the beta-annulus and the conserved D149 appear to be indispensable for the error-free assembly of T=3 particles. PMID- 15342252 TI - Role of metal ion-mediated interactions in the assembly and stability of Sesbania mosaic virus T=3 and T=1 capsids. AB - Sesbania mosaic virus (SeMV) capsids are stabilized by RNA-protein, protein protein and calcium-mediated protein-protein interactions. The removal of calcium has been proposed to be a prerequisite for the disassembly of the virus. The crystal structure of native T=3 SeMV capsid revealed that residues D146 and D149 from one subunit and Y205, N267 and N268 of the neighboring subunit form the calcium-binding site (CBS). The CBS environment is found to be identical even in the recombinant CP-NDelta65 T=1 capsids. Here, we have addressed the role of calcium and the residues involved in calcium co-ordination in the assembly and stability of T=3 and T=1 capsids by mutational analysis. Deletion of N267 and N268 did not affect T=3 or T=1 assembly, although the capsids were devoid of calcium, suggesting that assembly does not require calcium ions. However, the stability of the capsids was reduced drastically. Site-directed mutagenesis revealed that either a single mutation (D149N) or a double mutation (D146N-D149N) of SeMV coat protein affected drastically both the assembly and stability of T=3 capsids. On the other hand, the D146N-D149N mutation in CP-NDelta65 did not affect the assembly of T=1 capsid, although their stability was reduced considerably. Since the major difference between the T=3 and T=1 capsids is the absence of the N-terminal arginine-rich motif (N-ARM) and the beta-annulus from the subunits forming the T=1 capsids, it is possible that D149 initiates the N ARM-RNA interactions that lead to the formation of the beta-annulus, which is essential for T=3 capsid assembly. PMID- 15342253 TI - Crystal structure of calf spleen purine nucleoside phosphorylase with two full trimers in the asymmetric unit: important implications for the mechanism of catalysis. AB - The crystal structure of the binary complex of trimeric purine nucleoside phosphorylase (PNP) from calf spleen with the acyclic nucleoside phosphonate inhibitor 2,6-diamino-(S)-9-[2-(phosphonomethoxy)propyl]purine ((S)-PMPDAP) is determined at 2.3A resolution in space group P2(1)2(1)2(1). Crystallization in this space group, which is observed for the first time with a calf spleen PNP crystal structure, is obtained in the presence of calcium atoms. In contrast to the previously described cubic space group P2(1)3, two independent trimers are observed in the asymmetric unit, hence possible differences between monomers forming the biologically active trimer could be detected, if present. Such differences would be expected due to third-of-the-sites binding documented for transition-state events and inhibitors. However, no differences are noted, and binding stoichiometry of three inhibitor molecules per enzyme trimer is observed in the crystal structure, and in the parallel solution studies using isothermal titration calorimetry and spectrofluorimetric titrations. Presence of phosphate was shown to modify binding stoichiometry of hypoxanthine. Therefore, the enzyme was also crystallized in space group P2(1)2(1)2(1) in the presence of (S)-PMPDAP and phosphate, and the resulting structure of the binary PNP/(S)-PMPDAP complex was refined at 2.05A resolution. No qualitative differences between complexes obtained with and without the presence of phosphate were detected, except for the hydrogen bond contact of Arg84 and a phosphonate group, which is observed only in the former complex in three out of six independent monomers. Possible hydrogen bonds observed in the enzyme complexed with (S)-PMPDAP, in particular a putative hydrogen bonding contact N(1)-H cdots, three dots, centered Glu201, indicate that the inhibitor binds in a tautomeric or ionic form in which position N(1) acts as a hydrogen bond donor. This points to a crucial role of this hydrogen bond in defining specificity of trimeric PNPs and is in line with the proposed mechanism of catalysis in which this contact helps to stabilize the negative charge that accumulates on O(6) of the purine base in the transition state. In the present crystal structure the loop between Thr60 and Ala65 was found in a different conformation than that observed in crystal structures of trimeric PNPs up to now. Due to this change a new wide entrance is opened into the active site pocket, which is otherwise buried in the interior of the protein. Hence, our present crystal structure provides no obvious indication for obligatory binding of one of the substrates before binding of a second one; it is rather consistent with random binding of substrates. All these results provide new data for clarifying the mechanism of catalysis and give reasons for the non-Michaelis kinetics of trimeric PNPs. PMID- 15342254 TI - Solid-state 31P NMR spectroscopy of microcrystals of the Ras protein and its effector loop mutants: comparison between crystalline and solution state. AB - Cycling between a GTP bound "on" state and a GDP bound "off" state, guanine nucleotide-binding (GNB) proteins act as molecular switches. The switching process and the interaction with effectors, GTPase-activating proteins, and guanosine nucleotide-exchange factors is accompanied by pronounced conformational changes of the switch regions of the GNB proteins. The aim of the present contribution is to correlate conformational changes observed by liquid-state NMR with solid-state (31)P NMR data and with the results of X-ray crystallography. Crystalline wild-type Ras complexed with GTP analogs such as GppCH(2)p and GppNHp could be prepared. At low temperatures, two different signals were found for the gamma-phosphate group of GppNHp bound to wild-type Ras. This behavior indicates the existence of two different conformations of the molecule in the crystalline state as it is found in solution but not by X-ray crystallography. In contrast to the GppNHp complex, the two separate gamma-phosphate signals could not be observed for GppCH(2)p bound to wild-type Ras. However, an increasing linewidth at low temperature indicates the presence of an exchange process. The results obtained for the wild-type protein are compared with the behavior of GppNHp complexes of the effector loop mutants Ras(T35S) and Ras(T35A). These mutants prefer a conformation similar to the GDP bound "off" state. PMID- 15342255 TI - Crystal structure of the terminal oxygenase component of biphenyl dioxygenase derived from Rhodococcus sp. strain RHA1. AB - Biphenyl dioxygenase is the enzyme that catalyzes the stereospecific dioxygenation of the aromatic ring. This enzyme has attracted the attention of researchers due to its ability to oxidize polychlorinated biphenyls, which is one of the serious environmental contaminants. We determined the crystal structure of the terminal oxygenase component of the biphenyl dioxygenase (BphA1A2) derived from Rhodococcus strain sp. RHA1 in substrate-free and complex forms. These crystal structures revealed that the substrate-binding pocket makes significant conformational changes upon substrate binding to accommodate the substrate into the pocket. Our analysis of the crystal structures suggested that the residues in the substrate-binding pocket can be classified into three groups, which, respectively, seem to be responsible for the catalytic reaction, the orientation/conformation of the substrate, and the conformational changes of the substrate-binding pocket. The cooperative actions of residues in the three groups seem to determine the substrate specificity of the enzyme. PMID- 15342256 TI - Electrostatic potential of nucleotide-free protein is sufficient for discrimination between adenine and guanine-specific binding sites. AB - Despite sharing many common features, adenine-binding and guanine-binding sites in proteins often show a clear preference for the cognate over the non-cognate ligand. We have analyzed electrostatic potential (ESP) patterns at adenine and guanine-binding sites of a large number of non-redundant proteins where each binding site was first annotated as adenine/guanine-specific or non-specific from a survey of primary literature. We show that more than 90% of ESP variance at the binding sites is accounted for by only two principal component ESP vectors, each aligned to molecular dipoles of adenine and guanine. Projected on these principal component vectors, the adenine/guanine-specific and non-specific binding sites, including adenine-containing dinucleotides, show non-overlapping distributions. Adenine or guanine specificities of the binding sites also show high correlation with the corresponding electrostatic replacement (cognate by non-cognate ligand) energies. High correlation coefficients (0.94 for 35 adenine-binding sites and 1.0 for 20 guanine-binding sites) were obtained when adenine/guanine specificities were predicted using the replacement energies. Our results demonstrate that ligand-free protein ESP is an excellent indicator for discrimination between adenine and guanine-specific binding sites and that ESP of ligand-free protein can be used as a tool to annotate known and putative purine binding sites in proteins as adenine or guanine-specific. PMID- 15342257 TI - Revisiting preschoolers' living things concept: a microgenetic analysis of conceptual change in basic biology. AB - Many preschoolers know that plants and animals share basic biological properties, but this knowledge does not usually lead them to conclude that plants, like animals, are living things. To resolve this seeming paradox, we hypothesized that preschoolers largely base their judgments of life status on a biological property, capacity for teleological action, but that few preschoolers realize that plants possess this capacity. To test the hypothesis, we taught 5-year-olds one of four biological facts and examined the children's subsequent categorization of life status for numerous animals, plants, and artifacts. As predicted, a large majority of 5-year-olds who learned that both plants and animals, but not artifacts, move in goal-directed ways inferred that both plants and animals, but not artifacts, are alive. These children were considerably more likely to draw this inference than peers who learned that the same plants and animals grow or need water and almost as likely to do so as children who were explicitly told that animals and plants are living things and that artifacts are not. Results also indicated that not all biological properties are extended from familiar animals to plants; some biological properties are first attributed to plants and then extended to animals. PMID- 15342258 TI - Exploring automaticity in text processing: syntactic ambiguity as a test case. AB - A prevalent assumption in text comprehension research is that many aspects of text processing are automatic, with automaticity typically defined in terms of properties (e.g., speed and effort). The present research advocates conceptualization of automaticity in terms of underlying mechanisms and evaluates two such accounts, a computational-efficiency account (underlying computational processes become more efficient with practice) and a memory-based processing account (the underlying basis of processing shifts with practice, from computing interpretations to retrieving prior interpretations). In five experiments, short texts containing either an ambiguous or unambiguous syntactic structure were presented for multiple study trials. In both conditions, reading times in target regions decreased across trials, indicating automatization. Several findings supported the memory-based processing account (e.g., practice effects were largely item-specific, reading times were longer for ambiguous versus unambiguous sentences on early trials but converged on later trials) Some evidence was also found for a contribution of gains in computational efficiency (i.e., some item general practice effects were observed). Implications for research on automaticity and text processing are discussed. PMID- 15342259 TI - When learners surpass their models: the acquisition of American Sign Language from inconsistent input. AB - The present study examines the impact of highly inconsistent input on language acquisition. The American deaf community provides a unique opportunity to observe children exposed to nonnative language models as their only linguistic input. This research is a detailed case study of one child acquiring his native language in such circumstances. It asks whether this child is capable of organizing a natural language out of input data that are not representative of certain natural language principles. Simon is a deaf child whose deaf parents both learned American Sign Language (ASL) after age 15. Simon's only ASL input is provided by his late-learner parents. The study examines Simon's performance at age 7 on an ASL morphology task, compared with eight children who have native signing parents, and also compared with Simon's own parents. The results show that Simon's production of ASL substantially surpasses that of his parents. Simon's parents, like other late learners of ASL, perform below adult native signing criteria, with many inconsistencies and errors in their use of ASL morphology. In contrast, Simon's performance is much more regular, and in fact on most ASL morphemes is equal to that of children exposed to a native signing model. The results thus indicate that Simon is capable of acquiring a regular and orderly morphological rule system for which his input provides only highly inconsistent and noisy data. In addition, the results provide some insight into the mechanisms by which such learning may occur. Although the ASL situation is rare, it reveals clues that may contribute to our understanding of the human capacity for language learning. PMID- 15342260 TI - Uncovering the structure of a memorist's superior "basic" memory capacity. AB - After extensive laboratory testing of the famous memorist Rajan, Thompson, Cowan, and Frieman (1993) proposed that he was innately endowed with a superior memory capacity for digits and letters and thus violated the hypothesis that exceptional memory fully reflects acquired "skilled memory." We successfully replicated the empirical phenomena that led them to their conclusions. From additional analyses and new experiments, we found support for an alternative hypothesis, namely that Rajan's superior memory for digits was mediated by learned encoding techniques that he acquired during nearly a thousand hours of practice memorizing the mathematical constant pi. Our paper describes a general experimental approach for studying the structure of exceptional memory and how Rajan's unique structure is consistent with the general theoretical framework of long-term working memory (Ericsson and Kintsch, 1995). PMID- 15342261 TI - The developing constraints on parsing decisions: the role of lexical-biases and referential scenes in child and adult sentence processing. AB - Two striking contrasts currently exist in the sentence processing literature. First, whereas adult readers rely heavily on lexical information in the generation of syntactic alternatives, adult listeners in world-situated eye-gaze studies appear to allow referential evidence to override strong countervailing lexical biases (Tanenhaus, Spivey-Knowlton, Eberhard, and Sedivy, 1995). Second, in contrast to adults, children in similar listening studies fail to use this referential information and appear to rely exclusively on verb biases or perhaps syntactically based parsing principles (Trueswell, Sekerina, Hill, and Logrip, 1999). We explore these contrasts by fully crossing verb bias and referential manipulations in a study using the eye-gaze listening technique with adults (Experiment 1) and five-year-olds (Experiment 2). Results indicate that adults combine lexical and referential information to determine syntactic choice. Children rely exclusively on verb bias in their ultimate interpretation. However, their eye movements reveal an emerging sensitivity to referential constraints. The observed changes in information use over ontogenetic time best support a constraint-based lexicalist account of parsing development, which posits that highly reliable cues to structure, like lexical biases, will emerge earlier during development and more robustly than less reliable cues. PMID- 15342262 TI - Ganglioside GD1a increases the excitability of voltage-dependent sodium channels. AB - The effect of the negatively charged ganglioside GD1a, one of the major brain gangliosides [H. Beitinger, W. Probst, R. Hilbig, H. Rahmann, Seasonal variability of sialo-glycoconjugates in the brain of the Djungarian hamster (Phodopus sungorus). Comp. Biochem. Physiol., B 86 (1987) 377-384] on the function of brain derived BTX-modified voltage-dependent sodium channel was studied using the planar lipid bilayer system. Bilayers were formed either with a mixture of neutral phospholipids (4 phosphoethanolamine (PE):1 phosphocholine (PC)) alone or with one containing 6% of the disialoganglioside GD1a. The permeation and activation properties of the channels were measured in the presence of symmetrical 200 mM NaCl. We found that the single channel conductance was not affected by GD1a, whereas the steady-state activation curve displayed a hyperpolarizing shift in the presence of GD1a. Since the lipid distribution in these membranes is symmetrical, then the GD1a effect on sodium channels may result either from an induction of channel conformational changes or from an asymmetrical interaction between the channel (extracellular vs. intracellular channel aspect) and GD1a. Regardless of the mechanism, the data indicate that differences in ganglioside content in neuronal cells may contribute to the previously observed sodium channel functional variability within (soma, dentritic, axon hillock) and between neuronal cells as well as to excitability changes in those physiological and pathological conditions where changes in the neuronal ganglioside content occur. PMID- 15342263 TI - Serum S-100b protein as a biomarker for the assessment of neuroprotectants. AB - The study of biomarkers associated with stroke has proved to be of considerable utility. The astroglial protein S-100b is a candidate marker for cerebral tissue damage. We used a rat embolic model produced by injection of microspheres to demonstrate that serum S-100b is a useful biochemical marker for ischemic brain injury. Serum S-100b levels were significantly increased following microsphere injection, which was closely correlated with the development of brain edema. We found that structurally and mechanistically independent neuroprotective agents, such as 3-[2-[4-(3-chloro-2-methylphenylmethyl)-1-piperazinyl]ethyl]-5,6 dimethoxy-1-(4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate (DY 9760e), a novel calmodulin antagonist, and the N-methyl-d-aspartate (NMDA) receptor antagonist MK-801, are capable of attenuating increased serum S-100b levels and brain edema. In contrast, the hyperosmolar agent glycerol, which has no direct neuroprotective action, had little effect on serum S-100b levels, despite a significant decrease in brain water content. These results suggest that lowering of serum S-100b is mediated by neuroprotection against ischemic brain injury. Thus, serum S-100b reflects the extent of brain damage following cerebral ischemia and serves as a useful biomarker for the assessment of neuroprotectants. PMID- 15342264 TI - Hypoxia-ischemia affects erythropoietin and erythropoietin receptor expression pattern in the neonatal rat brain. AB - Erythropoietin (EPO), known for its role in erythroid differentiation, has been suggested to have non-hematopoietic functions in the brain, especially during development. In the present study, we investigated the expression of erythropoietin and erythropoietin receptor (EPOR) in the developing rat brain following hypoxia-ischemia. Seven-day-old rats underwent unilateral, permanent carotid artery ligation followed by 1 h of hypoxia, and their brains were examined immediately, 24 h or 4 days after hypoxia-ischemia. RT-PCR and Western blot analysis revealed that hypoxia-ischemia only marginally affected EPO expression. Immunohistochemical study of brains 4 days after hypoxia showed that 60 min of hypoxia (resulting in cortical infarction and severe neuronal loss in other regions) led to the increased EPO immunoreactivity, especially in the boundaries of the damaged cerebral cortex, associated with astrocytosis. In contrast, EPOR was dramatically upregulated within 24 h after hypoxia-ischemia. These results suggest that there is a rapid response of EPOR to the hypoxic ischemic stimulus, which seems to precede that of EPO, leading to the hypothesis that the EPO/EPOR system is implicated in the processes of neuroprotection from hypoxia-ischemia. PMID- 15342265 TI - Extrinsic tongue and suprahyoid muscle activities during mastication in freely feeding rabbits. AB - To evaluate the coordination of tongue and suprahyoid muscle activities during natural mastication, electromyograms (EMGs) of jaw-closer, jaw-opener, suprahyoid (mylohyoid, MH), tongue-retractor (styloglossus, SG) and tongue-protractor (genioglossus, GG) muscles were recorded as well as the jaw-movement trajectories in vertical and horizontal axes in awake rabbits. Each masticatory cycle had three components including the fast-closing (FC), slow-closing (SC) and opening (Op) phases. The duration of the SC phase was much longer during pellet chewing while the durations of the FC and Op phases were much shorter during pellet chewing than bread or banana chewing. The jaw movements during banana chewing had a small amplitude of lateral excursion and a large amplitude of gape as compared with those during pellet and bread chewing. The MH muscle exhibited double-peaked EMG bursts during the Op phase. The MH bursts in the late part of the Op phase were dominant on the non-chewing side during pellet and bread chewing. The SG muscle also exhibited double-peaked EMG bursts. During pellet and bread chewing, the SG bursts during the SC phase were significantly larger on the chewing side than the non-chewing side. These bursts were also dominant during pellet chewing as compared with banana chewing. There was little difference in the GG bursts between the chewing and non-chewing sides or among the foods. Our results suggest that patterns of the MH and SG muscle activity are affected by the peripheral inputs and/or chewing patterns while those of the GG muscle activity was less modulated regardless of the consistency of foods. PMID- 15342266 TI - Administration of MK-801 decreases c-Fos expression in the trigeminal sensory nuclear complex but increases it in the midbrain during experimental movement of rat molars. AB - Various studies reported c-Fos expression in the neurons in the trigeminal sensory nuclear complex (TSNC) following experimental tooth movement, which implies pain transmission to the central nervous system. Meanwhile, MK-801, a noncompetitive antagonist of N-methyl-D-aspartate (NMDA) receptors, was shown to markedly reduce the expression of c-Fos in the trigeminal subnucleus caudalis (Vc) following noxious stimulation but to enhance c-Fos expression markedly in other brain regions, i.e., the neocortex, dorsal raphe and thalamic nuclei. In the present study, we examined the nature of c-Fos expression in the brainstem including the TSNC and midbrain following administration of MK-801 and/or experimental movement of the rat molars. Twelve hours after the beginning of experimental tooth movement, c-Fos was expressed bilaterally in the superficial laminae of Vc (Vc I/II), dorsomedial areas of the trigeminal subnucleus oralis (Vodm) and rostro-dorsomedial areas of the trigeminal subnucleus oralis (Vor) with the ipsilaterally dominant distribution, but hardly in the periaqueductal gray (PAG), dorsal raphe nucleus (DR) and Edinger-Westphal nucleus (EW). Intraperitoneal administration of MK-801 (0.03, 0.3 and 3.0 mg/kg) prior to the onset of experimental tooth movement reduced c-Fos in the TSNC (Vc I/II, Vodm and Vor) but increased it in the nucleus raphe magnus (NRM), ventrolateral PAG (vl PAG), DR and EW. These results highly emphasize that during experimental tooth movement, a blockade of NMDA receptors induces neuronal suppression in the TSNC but increases neuronal activity in the descending antinociceptive system including the NRM, vl PAG, DR and EW. PMID- 15342267 TI - Chemically-induced long-term potentiation in rat motor cortex involves activation of extracellular signal-regulated kinase cascade. AB - The involvement of the extracellular signal-regulated kinase 1/2 (ERK1/2) cascade in long-lasting potentiation of synaptic transmission, induced by tetraethylammonium (TEA) or by elevated extracellular calcium concentration, was investigated in layer V horizontal connections within motor cortex in rat brain slices. Brief application of TEA (25 mM) resulted in a long-lasting potentiation of field potentials by 54+/-12%. A transient exposure of slices to elevated extracellular calcium (5 mM) induced long-lasting potentiation of responses reaching 30+/-8%. The induction of both forms of potentiation was prevented by the exposure of slices to inhibitors of the upstream activator of ERK 1/2, MEK (ERK kinase), U0126 (20 microM) and PD 98059 (50 microM). PhosphoERK2 immunoreactivity was transiently increased above baseline levels 15 min after termination of the exposure of slices to either TEA or elevated calcium concentration. Both forms of potentiation were partially occluded by Sp-adenosine 3',5'-cyclic monophosphorothioate triethylammonium salt (Sp-cAMPS; 100 microM), an activator of cAMP-dependent protein kinase (PKA), and they were blocked after preincubation with Rp-adenosine 3',5'-cyclic monophosphorothioate triethylammonium salt (Rp-cAMPS; 100 microM), a specific inhibitor of PKA activation by cAMP. It has previously been shown that TEA-induced potentiation represents a N-methyl-d-aspartate (NMDA) receptor-independent form of persistent synaptic enhancement, and, on the contrary, calcium-induced potentiation depends on NMDA receptors. Thus, the activation of PKA and the ERK1/2 cascade are required for two forms of chemically induced long-lasting increases of synaptic efficacy in slices of rat motor cortex. PMID- 15342268 TI - Postnatal maturation of cortical serotonin lateral asymmetry in gerbils is vulnerable to both environmental and pharmacological epigenetic challenges. AB - Long-term effects of postnatal differential rearing conditions and/or early methamphetamine (MA) application on serotonin (5-HT) fibre density were investigated in several cortical areas of both hemispheres of gerbils. The aim of this study was twofold: (1) Is the 5-HT fibre innervation of the cerebral cortex lateralised, and (2) if so, do postnatal environmental conditions and/or an early drug challenge interfere with development of 5-HT cerebral asymmetries? For that purpose, male gerbils were reared either under semi-natural or restricted environmental and social conditions, under both conditions once (on postnatal day 14) being treated with either a single dose of MA (50 mg/kg, i.p.) or saline. On postnatal day 110, 5-HT fibres were immunohistochemically stained and innervation densities quantified in prefrontal cortex, insular cortex, frontal cortex, parietal cortex, and entorhinal cortex. It was found that (1) 5-HT innervation in the cerebral cortex was clearly lateralised; (2) direction and extent of this asymmetry were not uniformly distributed over the different areas investigated; (3) both early methamphetamine challenge and rearing condition differentially interfered with adult 5-HT cerebral asymmetry; (4) combining MA challenge with subsequent restricted rearing tended to reverse the effects of MA on 5-HT cerebral asymmetry in some of the cortical areas investigated; and (5) significant responses in 5-HT cerebral asymmetry only occurred in prefrontal and entorhinal association cortices. The present findings suggest that the ontogenesis of cortical laterality is influenced by epigenetic factors and that disturbances of the postnatal maturation of lateralised functions may be associated with certain psychopathological behaviours. PMID- 15342269 TI - A distinct neurochemical profile in WKY rats at baseline and in response to acute stress: implications for animal models of anxiety and depression. AB - Wistar-Kyoto (WKY) rats exhibit hyperresponsive neuroendocrine and behavioral responses to stress that exceed normal controls and are especially prone to develop stress-induced depressive disorder. Pharmacological studies indicate altered serotonin (5-HT), norepinephrine (NE) and dopamine (DA) systems functioning in WKY rats, yet no attempt has been made to provide a comprehensive assessment of the neurochemical profile for WKY rats as compared to the outbred progenitor controls, Wistar rats. To this end, male, WKY and Wistar rats (N=6/group) were exposed to an acute forced-swim stress or were left untreated as controls. The prefrontal cortex (PFCtx), striatum, nucleus accumbens (NAS), and amygdala were assayed for levels of NE, DA and 5-HT, as well as major metabolites, by high-pressure liquid chromatography (HPLC) with electrochemical detection. In a separate experiment, designed to assess baseline and stress induced neuroendocrine activation, male, Wistar and WKY rats (N=6/group) were exposed to an acute forced-swim stress of 15 min or were left untreated as controls. Animals were killed immediately after the test (T=0), 30 min after the test (T=30) or 60 min after the test (T=60), and control animals were killed immediately after weighing. After decapitation, trunk blood was collected and plasma was isolated by centrifugation and analyzed for corticosterone by immunoassay. The neurochemical results demonstrate distinct patterns of baseline and stress-induced monoamine turnover in WKY rats, including alterations to DA and 5-HT turnovers in prefrontal cortex and nucleus accumbens, two critical brain areas implicated in anxiety, depression and drug reward. The neuroendocrine results indicate that WKY rats exhibited a sustained corticosterone response to acute stress, as compared to Wistar controls. Overall, these data are predicted to be useful for understanding the anxiety- and depressive-like behavioral phenotype exhibited by these animals and for increased understanding of the role genetic background in altering neurochemical function. PMID- 15342270 TI - Pressor response to CSF sodium in mice: mediation by a ouabain-like substance and renin-angiotensin system in the brain. AB - Intracerebroventricular (i.c.v.) infusion of sodium in rats increases cerebrospinal fluid (CSF) [Na], mimicking the effects of a high salt diet in salt sensitive strains and causing sympathetic hyperactivity and a pressor response that are mediated via both an endogenous brain ouabainlike substance (OLS) and the brain renin-angiotensin system (RAS). However, the concept that CSF sodium activates both the brain OLS and brain RAS to increase blood pressure has not been tested in any other species besides the rat. In the current study, it was established that continuous i.c.v. infusion of NaCl causes sustained increases in blood pressure and heart rate in both outbred (Swiss Webster, SW) and inbred (C57Bl/6) mouse strains. Subsequently, the mechanisms of the pressor effects were explored. In both SW and C57Bl/6, the i.c.v. administration of Fab fragments of an antibody with high affinity for ouabain and the OLS (Fab) abolished the pressor and tachycardic responses to i.c.v. sodium, as did the angiotensin II AT1 receptor antagonist losartan given i.c.v. In contrast, doses of NaCl, Fab and losartan that were effective i.c.v. were ineffective when given i.v. I.c.v. ouabain also caused the pressor and tachycardic responses, which were abolished by losartan (i.c.v.). In the reciprocal study, i.c.v. Fab had no effect on similar responses to i.c.v. angiotensin II. These studies demonstrate that the sustained blood pressure and heart rate responses caused by increases in CSF [Na] are mediated via both a brain OLS and the brain RAS. The RAS activation occurs downstream of the OLS effect. PMID- 15342271 TI - Taurine activates strychnine-sensitive glycine receptors in neurons of the rat inferior colliculus. AB - Taurine (Tau) is one of the most abundant free amino acids in the mammalian central nervous system. Whether the neurotransmission of the central auditory system is regulated or modulated by Tau is not clear. In the present study, we investigated the electrophysiological and pharmacological properties of Tau activated currents in acutely dissociated neurons of the rat inferior colliculus (IC) using whole cell patch clamp recordings. At a holding potential of -60 mV and under a condition of chloride equilibrium potential near 0 mV, Tau activated an inward current and its half-maximal activation concentration was equal to 0.37 mM. The measured reversal potential of Tau-activated currents was close to theoretical chloride equilibrium potential. The currents evoked by Tau at both low (1 mM) and high (10 mM) concentrations were almost completely inhibited by strychnine, a glycine receptor antagonist. The Tau-activated current, however, was not affected by bicuculline, a GABA(A) receptor antagonist. Tau at increased concentrations progressively reduced the current response to subsequent glycine application. At saturated concentrations, Tau-activated current and glycine activated current were mutually cross-desensitized by each other. These findings indicate that Tau activates glycine receptors in neurons of the rat IC and thus may have a functional role in regulating or modulating the neurotransmission of the central auditory system in mammals. PMID- 15342272 TI - Anti-apoptotic and pro-apoptotic effect of NEPP11 on manganese-induced apoptosis and JNK pathway activation in PC12 cells. AB - Neurite outgrowth-promoting prostaglandins (NEPPs), cyclopentenone prostaglandin derivatives, are found to be neurotrophic. These small organic compounds promote neurite outgrowth of PC12 cells and dorsal root ganglion explants in the presence of nerve growth factor, and prevent neuronal cell death of HT22 cells and cortical neurons induced by various stimuli. In this study, we examined whether NEPP11 prevents manganese-induced apoptosis of PC12 cells. NEPP11 (5 microM) attenuated manganese-induced DNA fragmentation by approximately 50%. In addition, NEPP11 partially prevented manganese-induced c-Jun phosphorylation and c-Jun N terminal kinase (JNK) phosphorylation determined by Western blotting. Inhibition of the JNK signaling pathway by NEPP11 appeared to be selective, because NEPP11 did not inhibit manganese-induced activation of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase1/2 (ERK1/2), MEK1/2 and p70 S6 kinase (p70S6K) in PC12 cells. In contrast, NEPP11 alone was toxic at higher concentrations (>10 microM) producing DNA fragmentation and activation of the JNK pathway. Molecular modifications of NEPP11 may strengthen its inhibitory effects on the JNK pathway while preventing its cytotoxicity, and thus may become a useful small molecule reagent for the treatment of manganese toxicity and other similar neurodegenerative processes. PMID- 15342273 TI - Activation of brain stem nuclei by improgan, a non-opioid analgesic. AB - Improgan is a compound developed from histamine antagonists which shows the pre clinical profile of a highly effective, non-opioid analgesic when administered into the rodent CNS. Pharmacological studies suggest that improgan activates descending pain-relieving circuits, but the brain and spinal sites of action of this drug have not been previously studied. Presently, the effects of intracerebral and intrathecal microinjections of improgan were evaluated on thermal nociceptive responses in rats. Improgan produced large, dose- and time related reductions in nociceptive responses following administration into the ventrolateral periaqueductal gray (PAG), the dorsal PAG, and the rostral ventromedial medulla (RVM). The drug had no measurable effects after injections into the caudate nucleus, basolateral amygdala, hippocampus, ventromedial hypothalamus, superior colliculi, ventrolateral medulla, or the spinal subarachnoid space. Inactivation of the RVM by muscimol microinjections completely attenuated antincociceptive responses produced by intraventricular improgan. These findings, taken with earlier results, show that, like opioids and cannabinoids, improgan acts in the PAG and RVM to activate descending analgesic systems. Unlike these other analgesics, improgan does not act in the spinal cord or in CNS areas outside of the brain stem. PMID- 15342274 TI - Reproductive activation of pine voles (Microtus pinetorum): examination of physiological markers. AB - We tested the hypothesis that the presence of an opposite-sex conspecific will result in time-related changes in measures of reproductive activation. We housed male-female pairs of pine voles together for 0, 2, 6, 12, or 24 h before collecting blood, reproductive organs and brains for immunocytochemical analysis of LHRH and c-fos. Control animals were never exposed to an opposite-sex conspecific. Following exposure to a male, there was a significant increase in uterine weight but not in LH levels. In males, there were no changes in peripheral indices of activation, i.e. LH levels, testes and seminal vesicle weights were not altered. Consistent with no change in circulating levels of LH, there was no change in LHRH immunoreactivity at any time. However, c-fos immunoreactivity was significantly greater in both males and females in the cingulate cortex and rostral bed nucleus of the stria terminalis (BNST) at 2 h, and in the caudal BNST at 2, 6 and 12 h. Similarly, c-fos immunoreactivity was increased in the rostral MPOA in both males and females at 2 and 6 h. However, in the caudal MPOA, there was a significant interaction between sex and time due to increased c-fos immunoreactivity in females only at 6 h. These results indicate that, in both male and female pine voles, exposure to an opposite-sex conspecific is sufficient to produce rapid, neural activation in brain areas known to be involved in reproductive activation and sexual behavior. This early activation did not occur in LHRH neurons. It is not known if this activation, particularly at early times, is due to reproductive activation or to the formation of pair bonds. PMID- 15342275 TI - Immunohistochemical characterisation and localisation of cannabinoid CB1 receptor protein in the rat vestibular nucleus complex and the effects of unilateral vestibular deafferentation. AB - CB1 receptor expression has been reported to be low in the brainstem compared with the forebrain, and low in the vestibular nucleus complex (VNC) compared with other regions in the brainstem. However, a frequent effect of cannabis is dizziness and loss of balance. This may be due to the activation of cannabinoid receptors in the central vestibular pathways. We used immunohistochemistry to study the distribution of CB1 receptor protein in the VNC, and Western blotting to measure CB1 receptor expression in the VNC following unilateral vestibular deafferentation (UVD); the hippocampal CA1, CA2/3 and dentate gyrus (DG) regions were also analysed for comparison. This study confirms a previous electrophysiological demonstration that CB1 receptors exist in significant densities in the VNC and are likely to contribute to the neurochemical control of the vestibular reflexes. Nonetheless, CB1 receptor expression did not change significantly in the VNC during vestibular compensation. In addition, despite some small but significant changes in CB1 receptor expression in the CA2/3 and the DG following UVD, in no case were these differences statistically significant in comparison to both control groups. PMID- 15342276 TI - Naltrexone attenuates cue- but not drug-induced methamphetamine seeking: a possible mechanism for the dissociation of primary and secondary reward. AB - The present study was aimed to clarify the role of the opioid system in the reinstatement of methamphetamine (METH)-seeking behavior in METH self administering rats. Following 12 days of self-administration of METH, the replacement of METH with saline resulted in a gradual decrease in lever press responses (extinction). Under extinction conditions, METH-priming or re-exposure to cues previously paired with METH infusion markedly increased the responses (reinstatement of drug-seeking). Naltrexone administered 30 min before re exposure to METH-associated cues attenuated reinstatement of drug-seeking behavior. On the other hand, administration of this antagonist had no effect on the reinstatement induced by METH-priming. We discussed these findings in relation with the dissociation of primary and secondary reward, suggesting that an opioid mechanism is responsible for this dissociation. Further, these results indicate the possibility of using naltrexone as an anti-relapse agent. PMID- 15342277 TI - Circadian and cAMP-dependent modulation of retinal cone cGMP-gated channels does not require protein synthesis or calcium influx through L-type channels. AB - Circadian oscillators of chicken retinal cone photoreceptors modulate the gating properties of cGMP-gated channels (CNGCs) such that they have a higher apparent affinity for cGMP during the subjective night. This effect is driven in part by cAMP, which acts through Erk MAP kinase to initiate a cascade leading to modulation of CGNCs. Here, we show that cAMP effects on the gating properties CNGCs persist when protein synthesis is blocked. The effects is cAMP also persist when calcium influx through L-type channels is blocked by nitrendipine. The mechanisms whereby cAMP modulates CNGCs therefore differ from those previously reported to underline regulation of melatonin synthesis and secretion. PMID- 15342278 TI - A single oral dose of geranylgeranylacetone attenuates kainic acid-induced seizures and neuronal cell death in rat hippocampus. AB - The present study evaluated the potential effect of geranylgeranylacetone (GGA), which is known as an antiulcer agent, against kainic acid (KA)-induced neurotoxicity. Pretreatment with a single oral GGA dose (800 mg/kg, 2 days before KA) significantly attenuated KA-induced seizures and cell death in rat hippocampus. These effects of GGA were prevented by the coinjection of MK801, a noncompetitive N-methyl-D-aspartate glutamate receptor antagonist, which indicates that the protection was indeed mediated by glutamate receptor activation. PMID- 15342279 TI - Leptin inhibits norepinephrine efflux from the hypothalamus in vitro: role of gamma aminobutyric acid. AB - Leptin, a hormone secreted by adipocytes, produces a number of central and neuroendocrine effects, the mechanisms behind which are not completely understood. Hypothalamic norepinephrine (NE) is involved in many of the neuroendocrine effects that are associated with leptin. Therefore, we hypothesized that leptin could affect hypothalamic NE activity to bring about its central and neuroendocrine effects. Because gamma aminobutyric acid (GABA) is known to affect the release of NE, we also tested the possibility that leptin induced changes in NE could be mediated through GABA. The mediobasal hypothalami from adult male rats were incubated in an in vitro incubation system for four consecutive incubation periods of 60 min each at 37 degrees C in Krebs Ringers Henseleit (KRH) solution in an atmosphere of 95% O(2) and 5% CO(2.) After determining the basal release, the hypothalami were challenged with 0, 0.1, 1 or 10 nm of leptin, bicuculline (a GABA-A receptor antagonist; 10 microM) and bicuculline (10 microM) +10 nM of leptin during the second incubation period. Residual effects of leptin were measured in the third incubation where tissues were incubated with KRH alone, and the viability of tissues was determined in the fourth incubation when tissues were exposed to high K(+) KRH. NE levels in the incubation medium were measured using high-performance liquid chromatography with electrochemical detection (HPLC-EC). Leptin inhibited NE efflux from the hypothalamus in a dose-dependent manner. Moreover, incubation of hypothalami with 10 nM of leptin and bicuculline, a completely blocked the leptin-induced decrease in NE efflux. These results demonstrate for the first time that leptin could act directly on the hypothalamus to inhibit NE efflux through GABA. It was concluded that leptin could probably produce its central and neuroendocrine effects by modulating NE and GABA levels in the hypothalamus. PMID- 15342280 TI - Alpha2-adrenoceptor modulation of long-term potentiation elicited in vivo in rat occipital cortex. AB - Pretreatment with the alpha(2)-adrenoceptor agonist clonidine (31.25, 62.5, or 125 microg/kg, i.p.) dose-dependently reduced long-term potentiation (LTP) elicited in vivo in the occipital cortex of anesthetized rats, whereas pretreatment with the alpha(2)-adrenoceptor antagonist yohimbine (0.133, 0.4, or 1.2 mg/kg, i.p.) increased neocortical LTP in a dose-dependent fashion. These effects could be related to the reported disruptive and facilitatory actions induced on memory formation by pretreatment with alpha(2)-adrenoceptor agonists and antagonists, respectively. PMID- 15342281 TI - Prevention of primary ventricular fibrillation in acute myocardial infarction with prophylactic lidocaine. AB - Primary ventricular fibrillation (VF) during an acute myocardial infarction (AMI) occurs with a high incidence and mortality rate with or without thrombolysis. The incidence varies from 2% to 19% depending on the definition of "primary." Primary VF in this study refers to fibrillation occurring in the absence of shock or pulmonary edema. Mortality rate, when primary VF occurs, is 2 to 4 times greater than when it does not. Prevention of VF has been impeded by the publication of the 1996 recommendations of the American Heart Association and American College of Cardiology against the use of prophylactic lidocaine based on meta-analysis studies implying toxicity. This observational study of 4,254 patients with AMI reports the incidence and mortality rates of primary VF over 32 years. Of the 4,254 patients, 4,150 received prophylactic lidocaine, and 104 patients did not receive prophylactic lidocaine due to the 1996 guidelines, after which administration of prophylactic lidocaine was governed by physician choice. The incidence of primary VF was 0.5% among the 4,150 who received prophylactic lidocaine and 10% among the 104 who did not (p <0.0001). Among the 4,150 receiving prophylactic lidocaine, sinoatrial block occurred in 0.5% and complete infranodal atrial ventricular block occurred in 0.2%, all secondary to the site of infarction (concurrent serum lidocaine levels were < 4 microg/ml). Asystole was an agonal rhythm in 4%; these patients had been off lidocaine for 48 hours. Mortality rates were 10.5% in patients without primary VF and 25% in patients with VF (p <0.001). Thus, prophylactic lidocaine markedly decreased the incidence of VF in 4,150 patients with AMI to 0.5% compared with trials before and after thrombolysis (2% to 19%) and with the 104 patients in this study who did not receive prophylactic lidocaine (10%). No lidocaine-induced sinoatrial or atrial ventricular block or asystole occurred. PMID- 15342282 TI - Accuracy of measurement of left ventricular volume and ejection fraction by new real-time three-dimensional echocardiography in patients with wall motion abnormalities secondary to myocardial infarction. AB - Three-dimensional echocardiography is an ideal tool for the measurement of left ventricular (LV) volume because no geometric assumptions about LV shape are needed. The introduction of new real-time 3-dimensional echocardiography (RT3DE) has allowed rapid acquisition of a 3-dimensional dataset with good image quality. The purpose of this study was to examine the accuracy of RT3DE for the measurement of LV volume and ejection fraction in patients with wall motion abnormalities by using quantitative gated single-photon emission computed tomography (QGSPECT) as a reference standard. The study population consisted of 25 consecutive patients with wall motion abnormalities who underwent LV volume measurement by 2-dimensional echocardiography and by QGSPECT. LV volume and ejection fraction by RT3DE were measured offline by using the average rotation method. In 23 of 25 patients (92%), it was possible to measure 3-dimensional volume with RT3DE. RT3DE correlated well with QGSPECT in the measurement of end diastolic volume and end-systolic volume (r = 0.97, mean difference 3.4 ml; r = 0.98, mean difference 2.0 ml, respectively), 2-dimensional echocardiography also correlated with QGSPECT but underestimated LV volume (r = 0.98, mean difference 21.1 ml; r = 0.98, mean difference 15.6 ml, respectively). Ejection fraction obtained by RT3DE had better agreement with that obtained by QGSPECT than that obtained by 2-dimensional echocardiography (r = 0.92, mean difference -0.2%; r = 0.89, mean difference -2.7%, respectively). RT3DE allows convenient and accurate estimation of LV volume and ejection fraction in patients with wall motion abnormalities. PMID- 15342283 TI - Prognostic value of exercise echocardiography in patients with classic angina pectoris. AB - The role of stress echocardiography in the prognostic evaluation of patients with angina pectoris is not well defined. This study included 437 patients (241 men and 196 women) with angina pectoris and a pretest probability of coronary artery disease (CAD) of > or = 0.7 who were referred for exercise echocardiography. No patient had a history of acute myocardial infarction or coronary revascularization. Mean age was 65 +/- 10 years. During a median follow-up of 2.7 years, hard cardiac events (cardiac death or nonfatal myocardial infarction) occurred in 19 patients and 53 patients underwent coronary revascularization. Event-free survival rates in patients with normal versus abnormal stress echocardiograms were 98% versus 83% at 1 year, 96% versus 75% at 3 years, and 87% versus 69% at 5 years, respectively. In a multivariate analysis of clinical, exercise stress, and echocardiographic parameters, independent predictors of hard cardiac events were Q waves on the electrocardiogram (chi-square 8.7, p = 0.003) and the presence of wall motion abnormalities during exercise in multivessel distribution (chi-square 5.3, p = 0.02). In an incremental model of clinical, exercise, and echocardiographic variables for the prediction of all cardiac events, the addition of echocardiographic data increased the chi-square of the model from 62 to 78 (p = 0.0003). Exercise echocardiography provides useful information in the risk stratification of patients with suspected CAD and a high pretest probability of CAD. Patients with normal exercise echocardiograms have a low event rate and therefore can be exempted from invasive procedures during the 3 years after a normal exercise echocardiogram. PMID- 15342284 TI - Effect of enalapril and losartan on cytokines in patients with stable angina pectoris awaiting coronary artery bypass grafting and their interaction with polymorphisms in the interleukin-6 gene. AB - Angiotensin-converting enzyme inhibitors and angiotensin receptor blockers may have anti-inflammatory actions, an effect that could explain some of their beneficial effects on cardiovascular events in clinical trials. Coronary artery bypass grafting (CABG) is associated with a systemic inflammatory response and provides a convenient model to examine the effects of such agents. Genetic polymorphisms may be important in influencing the expression of cytokines, such as interleukin-6 (IL-6). We randomized men awaiting CABG to treatment with enalapril, losartan, or control for 2 months before surgery. Systemic IL-6, IL-8, IL-10, and IL-1 receptor agonists were measured before and after surgery, and genotypes for the -174 G/C and -572 G/C IL-6 gene polymorphisms were determined. Total release of the IL-1 receptor agonist was decreased 29% by enalapril and 31% by losartan (adjusted p = 0.041). IL-6 was decreased 17% by enalapril and 20% by losartan. Subjects possessing the -174 GG genotype produced 20% more IL-6 (adjusted p = 0.029). In these high producers of IL-6, release of IL-6 was decreased 51% by enalapril (adjusted p = 0.001) and 32% by losartan (adjusted p = 0.068). Release of IL-10 was nonsignificantly decreased 26% by enalapril and 21% by losartan, whereas IL-8 was not detected. In conclusion, enalapril and losartan significantly decreased release of the IL-1 receptor agonist after CABG. Enalapril produced a highly significant decrease of 51% in the release of IL-6 in patients identified as high producers of IL-6 by the -174 G/C polymorphism, whereas losartan has a similar but less marked effect. The production of IL-6 in this setting is influenced by the -174 G/C polymorphism. PMID- 15342285 TI - Comparison of real-time myocardial contrast echocardiography for the assessment of myocardial viability with fluorodeoxyglucose-18 positron emission tomography and dobutamine stress echocardiography. AB - Little is known about the diagnostic value of real-time myocardial contrast echocardiography (MCE) for the assessment of myocardial viability. We compared the diagnostic accuracy of MCE with that of low-dose dobutamine stress echocardiography (DSE) and of combined technetium-99 sestamibi single-photon emission computed tomography and fluorodeoxyglucose-18 positron emission tomography and investigated whether quantitative assessment of myocardial blood flow could increase its diagnostic value. Cardiac imaging was performed with these 3 methods in 41 patients who had ischemic heart disease (ejection fraction < 40%) and were being considered for revascularization. Follow-up echocardiograms were obtained after 3 to 6 months in revascularized patients, and increased regional function served as a standard reference for assessment of myocardial viability. A control group of 25 patients who had no coronary artery disease underwent MCE to assess normal values of myocardial perfusion parameters. Recovery of myocardial function was predicted by MCE with a sensitivity of 86% and a specificity of 43%. Nuclear imaging was comparably sensitive (90%) and specific (44%), whereas low-dose DSE was similarly sensitive (83%) but more specific (76%). Normalization of myocardial signal intensity to that of the control group significantly increased the specificity of MCE from 43% to 64% and the accuracy from 73% to 81%. A combination of quantitative MCE and DSE provided the best diagnostic characteristics, with a sensitivity of 96%, a specificity of 63%, and an accuracy of 83%. Thus, MCE is useful for assessing myocardial viability. Normalization of contrast intensity to that of a reference control group is a valid approach for detection of myocardial viability and expands on information obtained from visual MCE and DSE. PMID- 15342286 TI - Coronary flow reserve of the angiographically normal left anterior descending coronary artery in patients with remote coronary artery disease. AB - Coronary artery disease (CAD) has been suggested to alter coronary flow reserve (CFR; the ratio between hyperemic and baseline coronary flow velocities) not only in territories supplied by stenotic arteries but also in angiographically normal, remote regions. However, few data exist regarding the left anterior descending (LAD) coronary artery as the normal index artery. The influence of remote CAD on CFR of the angiographically normal LAD was evaluated with transthoracic Doppler ultrasound to measure CFR in the LAD during 90 seconds of venous adenosine infusion (140 microg/kg/min) in 122 subjects who were assigned to 1 group; group 1 comprised 49 controls without angiographically detectable CAD, and group 2 consisted of 73 patients with an angiographically normal LAD and remote CAD. Group 2 was divided into 4 subgroups: 16 patients with previous remote percutaneous coronary intervention (group 2A); 13 patients with significant remote stenosis (group 2B); 23 patients with previous remote myocardial infarction and percutaneous coronary intervention (group 2C); and 21 patients with previous remote myocardial infarction but no percutaneous coronary intervention (group 2D). CFR in the LAD was not significantly different in groups 1 and 2 (3.08 +/- 0.61 and 3.03 +/- 0.69, respectively, p = NS). Decreased ejection fraction and increased wall motion score index in patients with remote CAD (p < 0.00001) and multivessel CAD did not affect CFR in the LAD (group 2A 3.18 +/- 0.77; group 2B 3.05 +/- 0.65; group 2C 3.07 +/- 0.79; group 2D 2.86 +/- 0.50, respectively; F = 0.63, p = NS). In conclusion, CFR of an angiographically normal LAD is preserved in patients with remote CAD, even in the presence of previous remote myocardial infarction and wall motion abnormalities. PMID- 15342287 TI - Comparison of outcomes of percutaneous coronary intervention of ostial versus nonostial narrowing of the major epicardial coronary arteries. AB - Outcomes of percutaneous coronary intervention (PCI) of the ostia of the major epicardial coronary arteries in the modern era of stenting have not been clearly defined. We evaluated data from all PCIs performed from 1998 to 2001 in the proximal segments of the major epicardial coronary arteries entered into a large cardiac database and compared ostial with nonostial PCI outcomes. Of 2,484 patients who underwent PCI of a proximal coronary artery (left anterior descending, left circumflex, or right coronary), 223 patients had ostial narrowing and 2,261 patients had proximal, nonostial narrowing. Baseline characteristics were similar between the 2 groups, except that patients with ostial narrowing tended to be older and have shorter narrowings than did patients with nonostial narrowings. Stenting occurred in 89% of all patients and was similar in patients with ostial or nonostial narrowings. Procedural success was the same for ostial and nonostial PCI (96% vs 95%, p = 0.95). One-year event-free survival rate was lower in patients who underwent ostial PCI (69% vs 80%, p = 0.0019), largely due to a greater need for repeat PCI (19% vs 10%, p <0.0001). Multivariate analysis showed that ostial location, age, angina class, and number of diseased vessels were independent predictors of the occurrence of cardiac events. PCI of ostial narrowings of the major epicardial coronary arteries was relatively safe. However, at 1 year, patients who underwent ostial PCI had an increased rate of repeat revascularization compared with patients who underwent nonostial, proximal PCI. PMID- 15342288 TI - Differential effect of two nicotinic acid preparations on low-density lipoprotein subclass distribution in patients classified as low-density lipoprotein pattern A, B, or I. AB - We tested the hypothesis that treatment with nicotinic acid results in a differential blood lipid response in subjects classified as having a low-density lipoprotein (LDL) pattern A or B. One hundred eighty hypercholesterolemic subjects were randomized to placebo (n = 61), immediate-release niacin (3,000 mg/day, n = 59), or extended-release niacin (1,500 mg/day, n = 60) for 14 weeks. Lipids and lipoprotein cholesterol were determined with enzymatic methods. LDL subclass distribution was determined with 2% to 16% polyacrylamide gradient gel electrophoresis. Extended- and immediate-release niacin had significant effects on the decrease of triglycerides, total cholesterol, LDL cholesterol, apoprotein B, lipoprotein(a), and apoprotein A-I and significantly increased high-density lipoprotein cholesterol. The 2 nicotinic acid compounds and doses significantly increased mean LDL peak particle diameter and percent distribution in large LDL I and IIa, with a significant decrease in small LDL IIIa, IIIb, and IVb. In patients with LDL pattern B compared with those with pattern A, extended-release niacin (1,500 mg/day) increased LDL peak particle diameter significantly more and decreased the percent distributions of small LDL IIIa, LDL IIIb, and LDL IVa significantly more. With 3,000 mg/day, immediate-release nicotinic acid in patients with LDL pattern B exhibited a significantly greater increase in LDL peak particle diameter and large LDL IIa and IIb and significantly greater decreases in small LDL IIIa, IIIb, and IVa compared with patients with pattern A. These differences in response between patients with LDL pattern A and those with pattern B were not reflected by changes in the standard lipid profile, including apoproteins A-I and B. Nicotinic acid has a significantly different effect on lipids and lipoprotein subclass distribution in subjects classified as having LDL subclass pattern A or B. Nicotinic acid has a significantly greater effect on the decrease of small LDL subclass distribution and increase in LDL peak particle diameter in pattern B versus pattern A. PMID- 15342289 TI - Safety and feasibility of using serial infusions of nesiritide for heart failure in an outpatient setting (from the FUSION I trial). AB - The Follow-Up Serial Infusions of Nesiritide pilot study was designed to assess the safety and tolerability of outpatient serial infusions of nesiritide in 210 patients with decompensated heart failure who were randomly assigned to usual care only or usual care plus weekly infusions of nesiritide at dosages of 0.005 or 0.01 microg/kg/min for 12 weeks. The mean age +/- SD of the entire population was 67 +/- 13 years; 70% were men, and 80% were white. Mean baseline serum creatinine levels were 1.8 +/- 0.7 mg/dl, and mean left ventricular ejection fraction was 0.28 +/- 0.15%. Diabetes mellitus was present in 106 patients (50%), and atrial arrhythmias were present in 100 patients (48%). A totalof 1,645 nesiritide infusions was administered; 11 (< 1%) were discontinued due to an adverse event. All treatment groups had a similar frequency of adverse events and experienced improvements in quality of life. Administration of nesiritide resulted in acute decreases in aldosterone and endothelin-1 concentrations. Although there were no statistically significant differences among groups by outcome, prospectively defined higher risk subgroups demonstrated significant decreases in cardiovascular events. These results demonstrate the safety and feasibility of administering nesiritide in an outpatient setting. Additional studies are needed to determine the effect of outpatient serial infusions of nesiritide on rates of morbidity and mortality in advanced heart failure. PMID- 15342290 TI - Evidence of viral infection in the myocardium of American and Japanese patients with idiopathic dilated cardiomyopathy. AB - Enteroviruses have been implicated in the pathogenesis of idiopathic dilated cardiomyopathy (IDC). Recently, the association of adenovirus or parvovirus with IDC has been reported. Viral infection in the myocardium of American and Japanese patients with end-stage IDC was evaluated. Myocardial specimens from 30 American patients with IDC and 47 Japanese patients with IDC were analyzed for the presence of cardiotropic viruses. The strand-specific detection of enteroviral ribonucleic acid (RNA) was performed to determine viral activity in hearts with IDC. Established reverse transcription-polymerase chain reaction (PCR) or PCR techniques were used to detect genomic sequences of influenza viruses, mumps virus, adenovirus, parvovirus, herpes simplex viruses, varicella-zoster virus, and Epstein-Barr virus. Enteroviral RNA was detected in 7 of the 30 American patients (23%) and in 15 of the 47 Japanese patients (32%). Minus-strand enteroviral RNA, an indicator of active enteroviral RNA replication, was demonstrated in 5 of 7 plus-strand-positive American patients (71%) and in 12 of 15 plus-strand-positive Japanese patients (80%). Sequence analysis revealed that the viruses detected were Coxsackie B viruses. No genomic sequences of other viruses were detected in the myocardium of either American or Japanese patients with IDC. Therefore, active group B Coxsackie virus RNA replication in the myocardium was demonstrated in a significant proportion of American and Japanese patients with end-stage IDC. There was no evidence of persistent infection by other viruses in hearts with IDC. Specific therapy should be designed for Coxsackie virus positive patients with IDC. PMID- 15342291 TI - Prophylactic lidocaine in acute myocardial infarction: resurface or reburial? PMID- 15342292 TI - William Peter Castelli, MD: a conversation with the editor [interview by William Clifford Roberts]. PMID- 15342293 TI - Comparison of long-term usefulness of clopidogrel therapy after the first percutaneous coronary intervention or coronary artery bypass grafting versus that after the second or repeat intervention. AB - Patients in the Clopidogrel for the Reduction of Events During Observation trial were subgrouped according to whether they underwent index percutaneous coronary artery revascularization procedures only or this procedure plus subsequent percutaneous or surgical revascularization procedures during 1-year follow-up. The relative risk reduction in ischemic events associated with up-front and long term clopidogrel compared with placebo was 2-fold greater in patients who required repeat revascularization procedures compared with those who did not (42.4% vs 21.7%). PMID- 15342294 TI - Sensitivity, specificity, positive predictive value, and negative predictive value of the dipyridamole sestamibi stress test in predicting graft occlusion or > or = 50% new native coronary artery disease in men versus women and in patients aged > or = 65 years versus < 65 years who had prior coronary artery bypass grafting. AB - The sensitivity, specificity, positive predictive value, and negative predictive value of the dipyridamole sestamibi stress test (DSST) in predicting graft obstruction or > or = 50% new native coronary artery disease (CAD) in 88 men compared with 56 women with prior coronary artery bypass surgery were 96% and 94% (p = NS), 30% and 67% (p = NS), 91% and 96 % (p = NS), 50% and 57% (p = NS), respectively. The sensitivity, specificity, positive predictive value, and negative predictive value of the DSST in predicting graft obstruction or > or = 50% new native CAD in 92 patients aged < 65 years were 95% and 95% (p = NS), 50% and 40% (p = NS), 96% and 87% (p = NS), and 43% and 67% (p = NS), respectively. PMID- 15342295 TI - Comparison of three-year outcomes after coronary stenting versus coronary artery bypass grafting in patients with multivessel coronary disease, including involvement of the left anterior descending coronary artery proximally (a subanalysis of the arterial revascularization therapies study trial). AB - The long-term effect of stents in patients with multivessel disease involving the proximal left anterior descending artery was investigated. At 3 years, there was no difference in the combined incidence of death, stroke, and myocardial infarction in either group, but the need for repeat revascularization was more frequent in the group with stenting than in the group with coronary artery bypass grafting. PMID- 15342296 TI - Comparison of mortality rates after acute myocardial infarction in smokers versus nonsmokers. AB - Patients who smoke paradoxically have favorable outcomes after acute myocardial infarctions compared with nonsmokers. However, after adjustment for age only, the decrease in all-cause mortality in the smoker population is explained by the smokers' generally younger age, with better prognoses due to their age. PMID- 15342297 TI - Magnitude and impact of treatment delays on weeknights and weekends in patients undergoing primary angioplasty for acute myocardial infarction (the cadillac trial). AB - In 2,082 patients in the CADILLAC trial, the outcomes of patients presenting during peak hours were compared with those presenting during peak hours (Monday to Friday 8a.m. to 8 p.m., n = 1,047, 51%) were compared with those of patients presenting during off-peak hours (weeknights from 8 p.m. to 8 a.m. and weekends, n = 989, 49%). Although treatment times to percutaneous coronary intervention (PCI) were delayed approximately 21 minutes, in patients with acute myocardial infarctions occurring on weeknights and weekends, this modest delay did not adversely affect procedural success, myocardial recovery, or survival after PCI. PMID- 15342298 TI - Comparison of effects of quinapril versus enalapril on vasoactive substances following acute myocardial infarction. AB - The true existence of a class effect in angiotensin-converting enzyme (ACE) inhibitors remains controversial. The present trial explored the effects of 2 ACE inhibitors after acute myocardial infarction and found no difference in endothelin-1 production but a greater increase in the production of total nitric oxide with quinapril than with enalapril. PMID- 15342299 TI - Mortality rates and hemorrhagic complications in asian-pacific islanders during treatment of acute myocardial infarction. AB - The management of acute myocardial infarction balances the benefits of antiplatelet and anticoagulant therapies against the risk for hemorrhage. The investigators report that Asian-Pacific islanders and patients with renal insufficiency have an increased rate of bleeding complications with hospitalized for acute myocardial infarction. PMID- 15342300 TI - Comparison of three-year outcomes in blacks versus whites with coronary heart disease following percutaneous coronary intervention. AB - There are limited data regarding the effect of race on survival after percutaneous coronary intervention (PCI) in the modern era. The investigators analyzed the impact of race on 3-year survival in 3,783 consecutive patients who underwent PCI in 1998 and 1999. PMID- 15342301 TI - Effect of chronic obstructive pulmonary disease on survival of patients with coronary heart disease having percutaneous coronary intervention. AB - There are limited data regarding the effect of chronic obstructive pulmonary disease (COPD) on the survival of patients with coronary artery disease. Prospectively developed and collected data elements on 4,284 consecutive patients who underwent percutaneous coronary intervention in 3 hospitals in 1998 and 1999 were pooled and analyzed. In-hospital major adverse cardiac outcomes were not different between groups. At 3-year follow-up, mortality for patients with COPD was 21% versus 9% for patients without COPD (log-rank p < 0.001). COPD was independently associated with a 2-fold increase in the hazard of long-term mortality (hazard ratio 2.146, 95% confidence interval 1.525 to 3.021, p < 0.001). PMID- 15342302 TI - Effect of simvastatin on endothelial function in cardiac syndrome X patients. AB - Patients with cardiac syndrome X with mild hypercholesterolemia were randomized to placebo (n = 20) or simvastatin 20 mg/day (n = 20). In the simvastatin group, there was a significant (26%; p < 0.0001) decrease in total cholesterol, a 38% (p < 0.0001) decrease in low-density lipoprotein cholesterol levels, and 7% a (p < 0.0001) increase in high-density lipoprotein cholesterol levels, without significant changes in triglyceride levels. Brachial artery flow-mediated dilation increased significantly (52% relative increase, p < 0.0001), and the time to > 1-mm ST-segment depression during stress testing was longer by the end of the study (p < 0.0001). PMID- 15342303 TI - Prevalence and determinants of breast arterial calcium in women at high risk of cardiovascular disease. AB - Calcium deposits in breast arteries are commonly seen on mammograms, and their frequency increases with age, especially after menopause. The investigators studied the prevalence of breast arterial calcium in 600 women at high risk for cardiovascular events and assessed whether classic cardiovascular risk factors are independent determinants of these calcifications. PMID- 15342304 TI - Comparative effects of carvedilol and amiodarone on conversion and recurrence rates of persistent atrial fibrillation. AB - Pretreatment with antiarrhythmic agents could improve cardioversion and recurrence rates in patients with persistent atrial fibrillation. In a prospective controlled trial, 145 patients were randomly assigned to treatment with carvedilol, amiodarone, or placebo for 4 weeks before electrical cardioversion. Although the 2 drugs had similar effects on cardioversion rates, amiodarone was superior in terms of sinus rhythm maintenance after conversion. PMID- 15342305 TI - Effectiveness of oral propafenone for the prevention of atrial fibrillation after coronary artery bypass grafting. AB - The efficacy and safety of oral propafenone for the prevention of atrial fibrillation (AF) were tested in a group of 293 patients who underwent coronary artery bypass grafting. Patients were randomized to placebo or 1 of 2 doses of propafenone. Patients treated with propafenone 675 mg/day compared with placebo had a nonsignificantly reduced incidence of AF and did not have a reduction in length of stay. Adverse events and discontinuations for untoward events were equal among the 3 groups. PMID- 15342306 TI - Effectiveness of catheter ablation for coexisting atrial fibrillation and atrial flutter. AB - In 79 consecutive patients (51 men and 28 women) with paroxysmal (n = 54) or persistent atrial fibrillation (AF) (n = 25) and typical, isthmus-dependent atrial flutter (AFl), pulmonary vein (PV) isolation and anatomically guided linear ablation of the right atrial isthmus was performed during the same procedure. After 208 +/- 331 days of follow-up, 42% remained free of AF and AFl, and a symptomatic lessening was reported by 77%. Paroxysmal AF (beta = 1.682, p = 0.008) and ablation of 3 or 4 PVs (beta = 1.830, p = 0.013) were independent predictors for arrhythmia-free survival. Combined catheter ablation of PVs and the right atrial isthmus for the treatment of patients with mixed AF and AFl is moderately effective in preventing early arrhythmia recurrence but leads to clinical improvement in most patients. PMID- 15342307 TI - N-terminal pro-brain natriuretic peptide reflects pulmonary capillary leakage in patients with acute dyspnea. AB - Natriuretic peptides have proved useful in the diagnosis of heart failure in patients presenting to the emergency department with shortness of breath. Dyspnea and orthopnea in heart failure are clinical expressions of pulmonary capillary congestion and leakage, which may be assessed by the percentage of pulmonary hemosiderin-laden macrophages (HLM) in induced sputum. We found a significant difference in the percentage of HLM present in sputum among patients with acute heart failure, patients with noncardiac dyspnea with ventricular dysfunction, and patients without heart failure (p = 0.008). N-terminal pro-brain natriuretic peptide (N-BNP) concentrations were also different among these 3 patient groups (p = 0.006). N-BNP concentrations were positively associated with the percentage of HLM in patients with acute dyspnea (r = 0.6; p < 0.0001). N-BNP, in addition to being a ventricular dysfunction marker, may reflect the severity of pulmonary capillary congestion and leakage in patients with acute shortness of breath. PMID- 15342308 TI - Effect of chronic heart failure on nuclear factor kappa B in peripheral leukocytes. AB - Nuclear factor kappa B (NF-kappaB) is a ubiquitous transcription factor activated by various stimuli that are implicated in the progression of chronic heart failure. Therefore, we examined the activation of NF-kappaB in peripheral leukocytes, the only nucleated cell population noninvasively accessible in patients with heart failure. In patients with stable heart failure with no obvious other reason for NF-kappaB activation, NF-kappaB was significantly activated. PMID- 15342309 TI - Proportion of patients in a congestive heart failure care management program meeting criteria for cardiac resynchronization therapy. AB - Among 1,138 patients in our congestive heart failure care management database, 37 (3.2%) met guideline criteria for cardiac resynchronization therapy. Advancing age was a predictor of prolonged QRS duration in this population, but gender, ejection fraction (EF) and cause of heart failure were not. There was a trend toward an inverse correlation between QRS duration and EF among patients with EF < or = 35%. PMID- 15342310 TI - Elevated B-type natriuretic peptide in asymptomatic men with chronic aortic regurgitation and preserved left ventricular systolic function. AB - Serum B-type natriuretic peptide (BNP) levels reflect myocardial strain and are known to be elevated in patients with heart failure. To determine if BNP levels are elevated in patients with aortic regurgitation, we measured BNP levels in patients with chronic asymptomatic aortic regurgitation and normal left ventricular systolic function. PMID- 15342311 TI - Weights of individual cusps in operatively-excised congenitally bicuspid stenotic aortic valves. AB - We weighed each cusp separately in 200 operatively excised stenotic congenitally bicuspid aortic valves. It was found in 152 patients (76%) that the cusps differed (by > 0.2 g) in weight and in 48 patients (24%) that the cusps were of similar (a < or = 0.2 g difference) weights. In 161 of the 200 patients (80%), raphes were present in 1 cusp; the raphe and nonraphe cusps differed in weight in 120 patients (74%), with the raphe cusps being larger in 89 patients (74%), and were of similar weights in 41 patients (26%). Of the 39 patients with raphes in neither cusp, 32 cusps (82%) differed in weight and 7 (18%) were of similar weights. The weight of each cusp differed more as the total valve weights increased. PMID- 15342312 TI - Weights of individual cusps in operatively-excised stenotic three-cuspid aortic valves. AB - We weighed each cusp separately in 260 operatively excised stenotic 3-cuspid aortic valves. All 3 cusps differed (by > 0.1 g) in weight in 69 patients (26%); all 3 cusps were similar (a < or = 0.1 g difference) in weight in 33 patients (13%); and 1 cusp differed from either of the other 2 cusps, which were similar in weight, in 158 patients (61%). The weight differences appeared to be caused by differing quantities of calcium on the aortic surfaces of the cusps. PMID- 15342313 TI - Coronary hemodynamics in patients with symptomatic hypertrophic cardiomyopathy. AB - The purpose of this study was to analyze coronary hemodynamics and determine a possible mechanism for microvascular dysfunction in patients with symptomatic hypertrophic cardiomyopathy (HC). Although patients with HC have been shown to have microvascular dysfunction and reduced coronary flow reserve (CFR), the mechanism by which this occurs is not well understood. We studied coronary hemodynamics in 8 patients with symptomatic HC and compared them with 8 matched controls. Compared with controls, patients with HC had higher coronary blood flow, lower coronary resistance, and lower CFR. Patients with HC also had abnormal phasic coronary flow characteristics. These results suggest that the reduction of CFR in patients with HC may be secondary to near maximal vasodilation of the microcirculation in the basal state. PMID- 15342314 TI - Implications and limitations of an abnormal fetal echocardiogram. AB - This study evaluates the accuracy of fetal echocardiograms in terms of anatomic diagnosis and predicted neonatal management over a 7-year period. Although an abnormal fetal echocardiogram is a highly reliable predictor of postnatal structural heart defects, challenges persist in the areas of conotruncal malformations, aortic arch, and pulmonary venous anomalies. PMID- 15342315 TI - Comparison of costs of intracardiac echocardiography and transesophageal echocardiography in monitoring percutaneous device closure of atrial septal defect in children and adults. AB - The cost of percutaneous device closure of atrial septal defect in children and adults using the Amplatzer Septal Occluder under intracardiac echocardiographic guidance was compared with that of transesophageal echocardiographic guidance. Hospital and physician charges incurred by 20 patients who had intracardiac echocardiographic guidance and 20 patients who had transesophageal echocardiographic guidance during atrial septal defect closure were compared. PMID- 15342316 TI - Repeatability of noninvasive surrogates of endothelial function. AB - We compared the repeatability of 2 ultrasonographic methods for endothelial function assessment at brachial artery-flow-mediated dilation (FMD) and post ischemic peak blood flow (PBF). Twenty healthy volunteers were examined twice within 10 days; coefficients of variation were 13.8% for PBF and 41.0% for FMD. PBF seems to be superior to FMD in terms of reproducibility. Consequently, smaller noninvasive studies of endothelial function can be designed utilizing PBF compared with FMD. PMID- 15342317 TI - Clinical significance of increased cardiac troponins T and I in participants of ultra-endurance events. AB - Of 105 asymptomatic finishers of endurance competitive events lasting several hours, increased blood concentrations of cardiac troponins T and I above the 99% upper reference values were found in 24 and 34 subjects, respectively; N-terminal pro-brain natriuretic peptide was also significantly increased. Within 3 months after the events, 21 troponin-positive participants underwent an extensive cardiac examination, which in all but 1 (critical coronary heart disease) revealed no signs of persistent cardiac damage. PMID- 15342318 TI - Long-term survival of childhood coronary artery disease in a patient severely affected with familial hypercholesterolemia. AB - This brief account of a patient severely affected with familial hypercholesterolemia and with coronary artery disease since age 13 reviews his therapy and survival into the fifth decade of life. This is heretofore unreported in phenotypically homozygous patients with familial hypercholesterolemia in the era before statin drugs, low-density lipoprotein apheresis, or their combination. PMID- 15342319 TI - Repetitive yawning associated with cardiac tamponade. AB - Yawning occurs frequently in daily life and is often attributed to boredom and fatigue. Rarely, it may be a symptom of serious underlying medical illness, primarily involving the central nervous system. We report a case of acute cardiac tamponade resulting from a large, malignant pericardial effusion. The patient manifested striking repetitive yawning that resolved immediately after pericardial drainage. An association between repetitive yawning and acute cardiac tamponade has not been previously described. PMID- 15342320 TI - A meta-analysis of the prevalence and usage of the personality disorder not otherwise specified (PDNOS) diagnosis. AB - The primary aim of the current study was to determine the prevalence and usage of the diagnosis of personality disorder not otherwise specified (PDNOS). As a secondary objective, this study explores the impact of assessment method and definition of PDNOS on observed prevalence. A meta-analysis is reported of 51 studies reporting separate rates for the overall prevalence of specific personality disorders and PDNOS. The best estimate of the absolute prevalence of PDNOS in patient samples is in the range of 8 to 13% and the best estimate of the relative prevalence of PDNOS (i.e., prevalence of PDNOS divided over the overall Axis II prevalence excluding PDNOS) is in the range of 21-49%. In structured interview studies PDNOS is the third most frequently used personality disorder diagnosis, whereas in nonstructured interview studies, PDNOS is often the single most frequently used diagnosis. PDE/IPDE yielded higher PDNOS prevalences than either the SCID or SIDP. Only slightly more than half of the studies provided an operational definition of PDNOS, and the most frequently occurring definition is "mixed" personality disorder. PDNOS would perhaps be the most frequently used diagnosis if it was used in a manner consistent with its definition in DSM-IV. The clinical relevance of future classification, assessment, and research could substantially benefit from validated, explicit, and uniform guidelines for the assessment of PDNOS. PMID- 15342321 TI - What's in a (neutral) face? Personality disorders, attachment styles, and the appraisal of ambiguous social cues. AB - Attachment theory suggests that borderline, avoidant, and schizoid personality disorders are differentially associated with anxious and avoidant attachment styles. The variations of insecure attachment in these disorders, in turn, might influence how others are perceived or appraised. To test these ideas, 176 college students completed questionnaires measuring attachment styles and personality disorder features, and each rated 10 emotionally neutral faces on 18 bipolar appraisal dimensions. As expected, borderline personality was associated with anxious attachment, avoidant personality was associated with anxious and avoidant attachment, and schizoid personality was weakly associated only with avoidant attachment. Furthermore, path analyses showed that borderline and avoidant personality features were associated with anxious attachment, which in turn related to negative face appraisals (e.g., tendencies to rate faces as less friendly and more rejecting). Discussion highlights the potential of simultaneously studying biases in social information processing and attachment disturbances among individuals with personality disorders. PMID- 15342322 TI - Reconstructing psychopathy: clarifying the significance of antisocial and socially deviant behavior in the diagnosis of psychopathic personality disorder. AB - A survey of clinical views suggests that the significance of antisocial and socially deviant behavior in the diagnosis of psychopathic personality disorder is unclear. To investigate this issue, we evaluated Psychopathy Checklist-Revised ratings (PCL-R; Hare, 1991) using structural equation modeling. One model, referred to as the measurement model, included PCL-R ratings related to antisocial behavior as primary symptoms of psychopathy; a second, referred to as the causal model, included the same PCL-R ratings as secondary symptoms or consequences. Compared to the measurement models, the causal model included more PCL-R items, was more parsimonious, and had equal or superior fit indices. These findings suggest that antisocial behavior is best viewed as a secondary symptom or consequence of psychopathy, In addition, the findings have important implications for future research and clinical-forensic practice, especially concerning the assessment of risk for criminality and violence. PMID- 15342323 TI - Time perception, impulsivity, emotionality, and personality in self-harming borderline personality disorder patients. AB - To investigate how time perception may contribute to the symptoms of self-harming Borderline Personality Disorder (BPD) patients, 19 self-harming BPD inpatients and 39 normal controls were given measures of time perception, impulsivity, personality, emotion, and BPD characteristics. A test sensitive to orbitofrontal cortex (OFC) function ("Frontal" Behavior Questionnaire) was also administered, as the OFC has been associated with impulsivity and time perception. BPD patients produced less time than controls, and this correlated with impulsiveness and other characteristics commonly associated with BPD. BPD patients were also less conscientious, extraverted, and open to experience, as well as more impulsive (self-report and behaviorally), emotional, neurotic, and reported more BPD characteristics, compared to controls. The results suggest that some of these core characteristics of BPD may be on a continuum with the normal population and, impulsivity in particular, may be related to time perception deficits (i.e., a faster subjective sense of time). Finally, BPD patients scored higher on the Frontal Behavior Questionnaire, suggesting that some symptoms of the BPD syndrome may be related to problems associated with the OFC. A control spatial working memory task (SWM) revealed that SWM deficits could not explain any of the BPD patients' poor performance. While impulsivity was correlated with time perception across all participants, emotionality, introversion, and lack of openness to experience were not. This suggests that different symptoms of the borderline personality syndrome may be separable, and therefore, related to different cognitive deficits, and potentially to different brain systems. This may have important implications for treatment strategies for BPD. PMID- 15342324 TI - Genetic and environmental structure of Cloninger's temperament and character dimensions. AB - The multivariate genetic and environmental structure of Cloninger's Temperament and Character Inventory (TCI) was investigated in a sample of 617 pairs of adolescent and young adult twins from Japan. Additive genetic factors accounted for 22% to 49% of the variability on all TCI temperament scales. Although the theory predicts lower heritability for the character scales, all character subscales had a substantial genetic contribution, and nonshared environmental influences accounted for the remainder. Multivariate genetic analyses showed that several subscales used to define one dimension shared a common genetic basis with subscales defining others. Using the degree of shared genetic influence as the basis to rearrange the TCI subscales into new dimensions, it was possible to create genetically independent scales. The implications for personality measurement, theory, and molecular genetic research are discussed. PMID- 15342325 TI - Temporal coherence of criteria for four personality disorders. AB - This study sought to investigate the coherence of changes observed in diagnostic criteria for borderline, schizotypal, obsessive-compulsive, and avoidant personality disorders. Five hundred, forty-nine patients were independently evaluated 2 years apart, and correlations of observed changes in each diagnostic criterion with changes in other criteria were examined to determine if there was within-syndrome consistency in these changes. The observed changes in criteria were consistent within syndrome (median alpha = 0.72 across 4 disorders), and reasonably specific to that syndrome relative to the other disorders. The results support the validity of these criterion sets as representing coherent syndromes. PMID- 15342326 TI - Personality traits in eating-disordered patients with and without self-injurious behaviors. AB - Although there is a high comorbidity of eating disorder and self-injurious behavior (SIB), no systematic research has focused on personality trait differences in patients with and without SIB. In this study, a Dutch adaptation of the NEO-FFI (Costa & McCrae, 1985, 1992) was completed by 178 female, eating disordered (ED) patients of whom 46.5% showed at least 1 form of SIB (e.g., cutting, burning, hair pulling). A subsample of 41 patients also completed the NEO-PI-R. Compared to patients without SIB, ED patients with SIB scored significantly higher on the Neuroticism scale and significantly lower on the Extraversion scale; on subtraits (facet scores) they appeared to be more anxious, more willing to please and less cheerful, efficient and ambitious. Personality traits were not associated with frequency or form of SIB or subtype of ED (except for impulsiveness). We also did not find a significant interaction effect between ED subtype and presence/absence of SIB. PMID- 15342327 TI - The ADP-IV questionnaire: differential validity and concordance with the semi structured interview. AB - The Assessment of DSM-IV Personality Disorders questionnaire (ADP-IV) is a self report measure of the DSM-IV Axis II personality disorders (PDs). The ADP-IV assesses for each DSM-IV criterion its typicality as well as the accompanying distress and impairment. This study investigates two important aspects of the construct validity of the ADP-IV: (a) the differential validity (i.e., the ability to differentiate between a sample of the general Flemish population ( n = 659) and a sample of psychiatric inpatients ( n = 487) with a high prevalence of clinical PD diagnoses, and patients with and without a PD in the psychiatric sample; (b) the convergent validity with the SCID-II semi-structured interview in a population of psychiatric inpatients ( n = 59). The results indicate a good differential validity: the dimensional scales and the categorical measures discriminated well between both groups and between patients with and without a PD diagnosis in the psychiatric sample. Concerning the concordance with the SCID-II, a decent level of agreement is exemplified by a correlation of.67 between the dimensional total scores of both instruments and by kappa coefficients for an "any" Axis II diagnosis at the.50 level. In conclusion, the results indicate that the ADP-IV is an efficient method for assessing PD in dimensional and categorical ways. PMID- 15342328 TI - Victim-victimizer relational dynamics as maintained by representational, defensive, and neurobiological functioning. AB - Cognitive-affective, defensive, and neurobiological aspects of victim-victimizer relational dynamics are illustrated. Observations of a 26-year-old mother and her 9-year-old son during a semistructured play task and projective and objective assessment data provide the sources of inference regarding how the parent and child's thematic structures are relationally expressed, maintain their psychopathology, and foster a victim-victimizer interactional cycle. By way of complementary interpersonal, psychological, and neurobiological processes, a victim-victimizer relational dynamic is maintained and intergenerationally transmitted. PMID- 15342329 TI - Cognitive enhancement therapy: a therapeutic treatment strategy for first-episode schizophrenia patients. AB - Cognitive enhancement therapy (CET) for first-episode schizophrenia combines supportive therapy with social skills and cognitive remediation training. It includes exercises aimed at ameliorating cognitive and negative symptoms in a purposefully motivational format. In this treatment model, the clinician takes an active role as coach, teacher, and therapist in order to engage patients in skills-enhancing work matched to levels of illness and abilities. The therapeutic alliance allows for a safe environment to utilize principles of errorless learning and positive reinforcement. Although this strategy is best implemented in groups, it can be adapted to the individual mode of treatment. Skills-building exercises are described in sufficient detail to be useful to clinicians working with patients in the first years of schizophrenia. PMID- 15342330 TI - Mourning, insight, and reduction of suicide risk in schizophrenia. AB - Literature on the assessment of suicide risk in individuals with schizophrenia is summarized, including the risk factors shared in common with the general population, illness-specific risk factors, and times of heightened risk in the course of the illness. Because depression emerges as a significant risk factor, it is differentiated from conditions that can mimic depression: mourning, aprosodia, and negative symptoms. Because insight or awareness of illness carries risk, as does the lack of insight, the psychological and neurocognitive components of impaired and of accurate insight are described. Finally, the role of mourning in the attainment of accurate, usable insight that can reduce suicidal risk is described. PMID- 15342331 TI - Aggression and guilt during mourning by parents who lost an infant. AB - The authors conducted in-depth interviews with 38 mothers and fathers who had lost an infant. The focus of the interview was aggression and guilt during mourning work. The participants felt strong shame after separation. Phenomena such as strong irrational guilt, aggression, and hesitation toward others were similar to feelings typical of the paranoid-schizoid position. However, mourners did not lose their sense of reality, continued to do daily chores, and kept taking care of others. In this period, mourners were in disintegration, similar to individuals in the paranoid-schizoid position. Shame may be the feeling in the residue of the paranoid-schizoid position, through which they felt persecuted by others, their partner, relatives, and God. Their aggression was strong, but generally their aggression and impulses came to be used constructively and they progressed to integration. At the moment their children died, mourners fluctuated between disintegration and integration. After a certain interval, fluctuation subsided or integration predominated. In times of fluctuation, containers were very important for mourners to move toward integration. Participants reported changes in their lifestyle after the loss of a child. Mental health professionals should view aggressive impulses among such people during mourning not as an obstacle, but rather as a means to move toward integration. The authors speculate that a psychotherapist can serve as a container for mourners. PMID- 15342332 TI - The influence of stigma on the sexual risk behavior of rural men who have sex with men. AB - Research investigating predictors of risky sexual behavior of rural MSM is sparse, even though the prevalence of HIV in rural areas has increased. This study explored two sets of predictors of 93 rural MSM's levels of risky sexual behavior: mental health variables and stigma emanating from men's family members, health care professionals, and people in the rural communities in which they live. Over 47% of the men were found to be at modified high to high risk. Logistic regression using a continuation logit model was used to test the relationship of the predictor variables and the four levels of risk. Findings indicate that self-esteem was predictive of the highest sexual risk behavior but not lower levels of risk. Stigma was predictive of modified high sexual risk when compared to low and no risk categories. No variables differentiated men at low risk from men at no risk. PMID- 15342333 TI - Behavioral and cognitive barriers to safer sex between men in steady relationships: implications for prevention strategies. AB - Steady partners are a major source of HIV infection among gay men. To better understand sexual risk taking in steady relationships, we examined characteristics of the first incident of unprotected anal intercourse (UAI) between steady male partners. We also examined cognitive barriers to safer sex by way of associating beliefs regarding UAI with protective behavior. Questionnaires assessing sexual behavior and related cognitions were completed by 324 gay men, aged 18-34. Of the men who had UAI with their steady partners, 55% (103/189) did so within the first 3 months of the relationship, and 46% did not discuss having UAI with their partner before it occurred. Analyses revealed that perceiving UAI as a symbol of trust and believing that the partner desired UAI were associated with less condom use but also with a higher likelihood that men established HIV negative seroconcordance and made sexual agreements (i.e., practiced negotiated safety). Perceiving UAI as more gratifying was associated with having risky UAI. Our findings suggest that interventions can make use of beliefs regarding trust and partner's desire for UAI to promote negotiated safety. In relationships where negotiated safety cannot be implemented, HIV prevention should regard the above beliefs, in particular the perception that UAI is more gratifying, as important barriers to safer sex. Furthermore, our findings regarding the early onset of risk in relationships emphasize how little time is at hand to prevent sexual risk before it occurs. One solution could be to target single gay men for promoting safer sex with future steady partners. PMID- 15342334 TI - Hookups or health promotion? An exploratory study of a chat room-based HIV prevention intervention for men who have sex with men. AB - This study sought to explore an online intervention that was initiated by an AIDS service organization to reduce sexual risk within a geographically-oriented chat room frequented by men who have sex with men (MSM). Analyses of bio lines, user profiles, and chat room discussions were used to understand and characterize the online prevention intervention. Six predominate discussion topics were identified, including (a) sexual risk reduction strategies (e.g., safer "barebacking"), (b) HIV testing options, (c) local alternatives for nonsexual social support, (d) referrals for youth, (e) resources related to "coming out," and (f) access to risk reduction materials and supplies. The perceived anonymity promoted by the Internet and the health educator's established relationship with the men in the chat room contributed to the success of this intervention. This analysis suggests that an online HIV/AIDS prevention intervention may reach MSM inaccessible in traditional venues typically used for intervention delivery. PMID- 15342335 TI - Correlates of high-risk sexual behavior among young men who have sex with men. AB - Data from a sample (n = 154) of young men who have sex with men (YMSM), aged 13 21, residing in five mid-sized cities in the midwestern United States were used to test the hypothesis that personality variables (impulsive decision making, sensation seeking, anxiety/depression, internalized homophobia) contribute uniquely to the prediction of unprotected anal sex beyond what is accounted for by other high-risk behaviors (i.e., substance use and number of sex partners) and partner status (having a dating partner). Results of hierarchical regression analyses indicated that number of reported anal sex partners and partner status were positively associated with self-reported frequency of unprotected anal sex. Beyond these factors, impulsive decision making significantly contributed to the predictive model. These findings indicate that designing effective prevention programming for YMSM should take into consideration relationship status and the impulsiveness of sexual decision making among members of the target group. PMID- 15342336 TI - Quality of study methods in individual- and group-level HIV intervention research: critical reporting elements. AB - To facilitate research synthesis and the identification of effective interventions, we reviewed and summarized critical reporting elements related to quality of study methods (QSM) specifically for individual- and group-level HIV intervention research. In developing these elements, we considered three sources of information: threats to validity, criteria and recommendations from review projects, and criteria and recommendations from published reviews relevant to HIV intervention research. Suggested QSM elements include, thoroughly describing intervention activities, using comparable outcome measures at preintervention and postintervention assessments, reporting data in detail for each study group, reporting participant refusal rates, including a comparison group, demonstrating study group comparability, clearly describing assignment to study groups, using appropriate statistical controls, collecting follow-up data from at least a 3 month postintervention period, reporting attrition in detail, and describing in detail whether the study sample size is adequate for detecting the expected effect size. Reporting on these QSM elements will assist in identifying effective behavioral interventions. PMID- 15342337 TI - Psychosocial antecedents of injection risk reduction: a multivariate analysis. AB - The AIDS risk reduction model (ARRM) predicts decreased occurrence of AIDS risk behavior through a three-stage process: (a) perceiving one's behavior as risky and recognizing one's skills to reduce the behavior, (b) forming an intention to change behavior, and (c) acting on that intention. We used the ARRM to test a stage-based longitudinal structural equation model of the impact of intention to reduce injection risk behavior on subsequent behavior change in a sample of 294 HIV-negative opiate-addicted individuals in treatment. Intended risk reduction and continuous participation in treatment significantly predicted less injection risk behavior. Greater self-efficacy, less baseline risk behavior, less perceived susceptibility to AIDS, and greater fear of AIDS predicted intention to reduce risk. Leverage points for change are discussed. PMID- 15342338 TI - HIV risk behaviors, knowledge, and prevention education among offenders under community supervision: a hidden risk group. AB - Numerous studies have established that incarcerated populations are at substantial risk for HIV infection. In response, many jails and prisons have increased HIV prevention and related services. However, although twice as many offenders are under community supervision as are incarcerated at any given time, HIV prevention needs have been largely ignored among probationers and parolees, and little is known about their HIV risk behaviors or HIV prevention needs. Compared with inmates, probationers and parolees have substantially greater opportunities to engage in HIV risk behaviors. In the present study, we describe HIV risk behaviors, knowledge, and prevention education experiences of probationers and parolees in New York City. We find that probationers and parolees have high rates of unprotected sex, and limited current exposure to effective HIV education and prevention interventions. Probation and parole departments need to improve HIV training for officers and make HIV risk reduction services more available. PMID- 15342339 TI - Native, not nitrated, cytochrome c and mitochondria-derived hydrogen peroxide drive osteoclast apoptosis. AB - Two unresolved aspects of the role of mitochondria-derived cytochrome c in apoptosis are whether there is a separate pool of cytochrome c within mitochondria that participates in the activation of apoptosis and whether a chemically modified cytochrome c drives apoptosis. These questions were investigated using osteoclasts, because they are rich in mitochondria and because osteoclast apoptosis is critical in bone metabolism regulation. H(2)O(2) production was increased during culture, preceding cytochrome c release; both processes occurred anterior to apoptosis. With the addition of a mitochondrial uncoupler, H(2)O(2) production and apoptosis were blocked, indicating the prominent role of mitochondria-derived H(2)O(2). Trapping H(2)O(2)-derived hydroxyl radical decreased apoptosis. Cytosolic cytochrome c was originated from a single mitochondrial compartment, supporting a common pool involved in respiration and apoptosis, and it was chemically identical to the native form, with no indication of oxidative or nitrative modifications. Protein levels of Bcl 2 and Bc-xL were decreased before apoptosis, whereas expression of wild-type Bcl 2 repressed apoptosis, confirming that cytochrome c release is critical in initiating apoptosis. Cytosolic cytochrome c participated in activating caspase-3 and -9, both required for apoptosis. Collectively, our data indicate that the mitochondria-dependent apoptotic pathway is one of the major routes operating in osteoclasts. PMID- 15342340 TI - Serum and glucocorticoid-regulated kinase Sgk1 inhibits insulin-dependent activation of phosphomannomutase 2 in transfected COS-7 cells. AB - Serum- and glucocorticoid-regulated kinase (Sgk1) is considered to be an essential convergence point for peptide and steroid regulation of ENaC-mediated sodium transport. We tried to identify molecular partners of Sgk1 by yeast two hybrid screening. Yeast two-hybrid screening showed a specific interaction between Sgk1 and phosphomannomutase (PMM)2, the latter of which is an enzyme involved in the regulation of glycoprotein biosynthesis. The interaction was confirmed in intact cells by coimmunoprecipitation and colocalization detected using confocal microscopy. We were then able to demonstrate that Sgk1 phosphorylated PMM2 in an in vitro assay. In addition, we found that the enzymatic activity of PMM2 is upregulated by insulin treatment and that Sgk1 completely inhibits PMM2 activity both in the absence and in the presence of insulin stimulation. These data provide evidence suggesting that Sgk1 may modulate insulin action on the cotranslational glycosylation of glycoproteins. PMID- 15342341 TI - Synergistic amplification of beta-amyloid- and interferon-gamma-induced microglial neurotoxic response by the senile plaque component chromogranin A. AB - Activation of the microglial neurotoxic response by components of the senile plaque plays a critical role in the pathophysiology of Alzheimer's disease (AD). Microglia induce neurodegeneration primarily by secreting nitric oxide (NO), tumor necrosis factor-alpha (TNFalpha), and hydrogen peroxide. Central to the activation of microglia is the membrane receptor CD40, which is the target of costimulators such as interferon-gamma (IFNgamma). Chromogranin A (CGA) is a recently identified endogenous component of the neurodegenerative plaques of AD and Parkinson's disease. CGA stimulates microglial secretion of NO and TNFalpha, resulting in both neuronal and microglial apoptosis. Using electrochemical recording from primary rat microglial cells in culture, we have shown in the present study that CGA alone induces a fast-initiating oxidative burst in microglia. We compared the potency of CGA with that of beta-amyloid (betaA) under identical conditions and found that CGA induces 5-7 times greater NO and TNFalpha secretion. Coapplication of CGA with betaA or with IFNgamma resulted in a synergistic effect on NO and TNFalpha secretion. CD40 expression was induced by CGA and was further increased when betaA or IFNgamma was added in combination. Tyrphostin A1 (TyrA1), which inhibits the CD40 cascade, exerted a dose-dependent inhibition of the CGA effect alone and in combination with IFNgamma and betaA. Furthermore, CGA-induced mitochondrial depolarization, which precedes microglial apoptosis, was fully blocked in the presence of TyrA1. Our results demonstrate the involvement of CGA with other components of the senile plaque and raise the possibility that a narrowly acting agent such as TyrA1 attenuates plaque formation. PMID- 15342342 TI - Canonical transient receptor potential TRPC7 can function as both a receptor- and store-operated channel in HEK-293 cells. AB - Previous studies on the activation mechanism of canonical transient receptor potential (TRPC) channels have often produced conflicting conclusions. All seven have been shown to be activated by phospholipase C (PLC)-coupled receptors, but TRPC1, TRPC2, TRPC3, TRPC4, TRPC5, and TRPC7 have also been proposed to function as store-operated channels.(1)1Although PLC activation inevitably leads to activation of store-operated channels, in this report when we refer to PLC activated channels, we mean those channels that are specifically activated by PLC independently of store depletion. In the case of TRPC3, the expression environment and the expression level appear to determine the mode of regulation. Evidence of a close structural relative of TRPC3, TRPC7, has been presented that this channel is activated by receptor activation or by store depletion. On the basis of previous findings for TRPC3, we reasoned that subtle differences in structure or expression conditions might account for the apparent distinct gating mechanisms of TRPC7. To reexamine the mode of activation of TRPC7, we stably and transiently transfected human embryonic kidney (HEK)-293 cells with cDNA encoding for human TRPC7. We examined the ability of a PLC-activating agonist and an intracellular Ca(2+) store-depleting agent to activate these channels. Our findings demonstrate that when transiently expressed in HEK-293 cells, TRPC7 forms channels that are activated by PLC-stimulating agonists, but not by Ca(2+) store depletion. However, when stably expressed in HEK-293 cells, TRPC7 can be activated by either Ca(2+) store depletion or PLC activation. To our knowledge, this is the first demonstration of a channel protein that can be activated by both receptor- and store-operated modes in the same cell. In addition, the results reconcile the apparently conflicting findings of other laboratories regarding TRPC7 regulation. PMID- 15342343 TI - Barrier role of actin filaments in regulated mucin secretion from airway goblet cells. AB - Airway goblet cells secrete mucin onto mucosal surfaces under the regulation of an apical, phospholipase C/G(q)-coupled P2Y(2) receptor. We tested whether cortical actin filaments negatively regulate exocytosis in goblet cells by forming a barrier between secretory granules and plasma membrane docking sites as postulated for other secretory cells. Immunostaining of human lung tissues and SPOC1 cells (an epithelial, mucin-secreting cell line) revealed an apical distribution of beta- and gamma-actin in ciliated and goblet cells. In goblet cells, actin appeared as a prominent subplasmalemmal sheet lying between granules and the apical membrane, and it disappeared from SPOC1 cells activated by purinergic agonist. Disruption of actin filaments with latrunculin A stimulated SPOC1 cell mucin secretion under basal and agonist-activated conditions, whereas stabilization with jasplakinolide or overexpression of beta- or gamma-actin conjugated to yellow fluorescent protein (YFP) inhibited secretion. Myristoylated alanine-rich C kinase substrate, a PKC-activated actin-plasma membrane tethering protein, was phosphorylated after agonist stimulation, suggesting a translocation to the cytosol. Scinderin (or adseverin), a Ca(2+)-activated actin filament severing and capping protein was cloned from human airway and SPOC1 cells, and synthetic peptides corresponding to its actin-binding domains inhibited mucin secretion. We conclude that actin filaments negatively regulate mucin secretion basally in airway goblet cells and are dynamically remodeled in agonist stimulated cells to promote exocytosis. PMID- 15342344 TI - Introduction of Dr. Alfred M. Cohen. PMID- 15342345 TI - Society of surgical oncology presidential address: friendships, partnerships, and teams--keys to academic success. PMID- 15342346 TI - Honoring Blake Cady, MD: Society of Surgical Oncology President, 1988-1989. PMID- 15342347 TI - Models of breast cancer growth and investigations of adjuvant surgical oophorectomy. AB - Clinical observations of the natural history of breast cancer and its response to a variety of therapeutic interventions have contributed to changing concepts about the growth and metastatic spread of this disease. Increased attention has been given to tumor cell dormancy and the occurrence of greatly delayed metastatic disease development, which has been important to rethinking therapy. Although gene profiling of breast tumors recently has highlighted the importance of individual tumor characteristics in patients' prognosis, considerable data also support the concept of breast cancer as a problem of macro- and microenvironmental regulatory imbalance and dynamic chaos. Observations of unexpectedly large survival benefits from adjuvant surgical oophorectomy done in the luteal phase of the menstrual cycle in premenopausal women are consistent with an interpretation that extratumoral interactions in the host environment are important in prognosis. These observations also suggest that a treatment paradigm shift from an exclusive focus on cell kill and specific tumor cell molecular targets to one focused also on broad host regulatory control may be useful. Clinical trials and laboratory mechanistic investigations based on these data and observations can determine the potential impact of therapeutic interventions targeting host system macro and micro tumor cell environments. PMID- 15342348 TI - Long-term survival after an aggressive surgical approach in patients with breast cancer hepatic metastases. AB - BACKGROUND: Metastatic breast cancer is generally believed to be associated with a poor prognosis. Therapeutic advances over the past two decades, however, have resulted in improved outcomes for selected patients with limited metastatic disease. METHODS: Between March 1991 and October 2002, 31 patients had hepatic resection for breast cancer metastases limited to the liver. Clinical and pathologic data were collected prospectively from breast and hepatobiliary databases. RESULTS: Median age of patients was 46 years (range, 31 to 70). Liver metastases were solitary in 20 patients and multiple in 11 patients. Median size of the largest liver metastasis was 2.9 cm (range, 1 to 8). Major liver resections (three or more segments resected) were performed in 14 patients, whereas minor resections (fewer than three segments resected) with or without radiofrequency ablation (RFA) were performed in 17 patients. No postoperative mortality occurred. Of the 31 patients, 27 (87%) received either preoperative or postoperative systemic therapy as treatment for metastatic disease. The median survival was 63 months; a single patient died within 12 months of hepatic resection. The overall 2- and 5-year survival rates were 86% and 61%, respectively, whereas the 2- and 5-year disease-free survival rates were 39% and 31%, respectively. No treatment- or patient-specific variables were found to correlate with survival rates. CONCLUSIONS: In selected patients with liver metastases from breast cancer, an aggressive surgical approach is associated with favorable long-term survival. Hepatic resection should be considered a component of multimodality treatment of breast cancer in these patients. PMID- 15342349 TI - Dr. Herbert Lumley Snow, MD, MRCS (1847-1930): the original champion of elective lymph node dissection in melanoma. AB - Herbert Snow (1847-1930), a London surgeon with a particular interest in melanoma, was a controversial proponent of anticipatory gland excision well before acceptance of elective lymph node dissections. This article describes the work of Snow within a wider historical context. PMID- 15342350 TI - The utility of frozen section evaluation for follicular thyroid lesions. PMID- 15342351 TI - The removal of regional nodes even when "pathologically negative" for long-term survival should be discussed cautiously. PMID- 15342352 TI - Expression, regulation, and immunolocalization of putative homeodomain transcription factor 1 (PHTF1) in rodent epididymis: evidence for a novel form resulting from proteolytic cleavage. AB - PHTF1 is an 84-86-kDa membrane protein found in the endoplasmic reticulum of male germ cells in rodents. There are no evident signs of PHTF1 in the spermatozoa released into the lumen of the seminiferous tubules but PHTF1 is present in the epididymal epithelium. Characterization of the epididymal Phtf1 messenger by Northern blot and reverse transcription-PCR identified a 3-kilobase transcript in the epididymis, similar to that previously reported in the testis. The transcript is present in the proximal part of the epididymis and it appears when the rats reach 4 wk of age. Through immunofluorescence analysis, PHTF1 was localized in the principal cells of the initial segment and the caput epididymis. Colocalization with different markers indicated PHTF1 is in the endoplasmic reticulum saccules applied to the trans face of the Golgi system. Western blot analyses revealed a shorter form of the protein--about 56-kDa versus the 84-kDa form found in the testis. Using the canine epididymal cell line CIM 20, transfected by N- and C-terminal myc-tagged PHTF1, we demonstrated that the 56 kDa epididymal form could result from proteolytical processing. PMID- 15342353 TI - The deubiquitinating enzyme mUBPy interacts with the sperm-specific molecular chaperone MSJ-1: the relation with the proteasome, acrosome, and centrosome in mouse male germ cells. AB - The mouse USP8/mUBPy gene codifies a deubiquitinating enzyme expressed preferentially in testis and brain. While the ubiquitin-specific processing proteases (UBPs) are known to be important for the early development in invertebrate organisms, their specific functions remain still unclear in mammals. Using specific antibodies, raised against a recombinant mUBPy protein, we studied mUBPy in mouse testis. The mUBPy is expressed exclusively by the germ cell component and is maintained in epididymal spermatozoa. The enzyme is functionally active, being able to detach ubiquitin moieties from endogenous protein substrates. Protein interaction assays showed that sperm UBPy interacts with MSJ 1, the sperm-specific DnaJ protein evolutionarily conserved for spermiogenesis. Immunocytochemistry revealed that mUBPy shares with MSJ-1 the intracellular localization during spermatid cell differentiation; intriguingly, we show here that the proteasomes also locate in mUBPy/MSJ-1-positive sites, such as the cytoplasmic surface of the developing acrosome and the centrosomal region. These colocalization sites are maintained in epididymal spermatozoa. The demonstration of a protein interaction between a deubiquitinating enzyme and a molecular chaperone and the documentation on the proteasomes in both differentiating and mature mouse male germ cells suggest that members of the chaperone and ubiquitin/proteasome systems could cooperate in the fine control of protein quality to yield functional spermatozoa. PMID- 15342354 TI - Involvement of the cytoskeleton in oxytocin secretion by cultured bovine luteal cells. AB - A number of substances have been implicated in the regulation of oxytocin (OT) secretion from bovine corpus luteum in vivo. However, isolated bovine luteal cells cultured in a monolayer lose the ability to secrete OT in response to stimulatory substances. The present study investigated how cell-to-cell contact and the cytoskeleton affect OT secretion by isolated bovine luteal cells. In experiment 1, bovine midluteal cells (Days 8-12 of the estrous cycle) were stimulated with prostaglandin F2alpha (PGF2alpha; 1 microM), noradrenaline (NA; 10 microM), or growth hormone (GH; 5 nM) in two culture systems: In one system, cell monolayers were incubated in 24-well culture plates, and in the other system, aggregates of cells were incubated in glass tubes in a shaking water bath. The cells cultured in a monolayer underwent considerable spreading and showed a variety of shapes, whereas the cells cultured in glass tubes remained fully rounded during the experimental period and soon formed aggregates of cells. Although PGF2alpha, NA, and GH did not stimulate OT secretion by the monolayer cells, all tested substances stimulated OT secretion by the aggregated cells (P < 0.01). In experiment 2, the monolayer cells were pre-exposed for 1 h to an antimicrofilament agent (cytochalasin B; 1 microM) or two antimicrotubule agents (colchicine or vinblastine; 1 microM) before stimulation with PGF2alpha, NA, or GH. Although PGF2alpha, NA, and GH did not stimulate OT secretion by the monolayer cells in the presence of colchicine or vinblastine, they all stimulated OT secretion in the presence of cytochalasin B (P < 0.001). The overall results show that OT secretion by bovine luteal cells depends on microfilament function and cell shape. Moreover, the aggregate culture system that allows three dimensional, cell-to-cell contact seems to be a good model for studying OT secretion by isolated bovine luteal cells. PMID- 15342355 TI - Tyrosine phosphorylation of the a kinase anchoring protein 3 (AKAP3) and soluble adenylate cyclase are involved in the increase of human sperm motility by bicarbonate. AB - Mammalian testicular spermatozoa are immotile, thus, to reach the oocyte, they need to acquire swimming ability under the control of different factors acting during the sperm transit through the epididymis and the female genital tract. Although bicarbonate is known to physiologically increase motility by stimulating soluble adenylate cyclase (sAC) activity of mammalian spermatozoa, no extensive studies in human sperm have been performed yet to elucidate the additional molecular mechanisms involved. In this light, we investigated the effect of in vitro addition of bicarbonate to human spermatozoa on the main intracellular signaling pathways involved in regulation of motility, namely, intracellular cAMP production and protein tyrosine phosphorylation. Bicarbonate effects were compared with those of the phosphatidyl-inositol-3 kinase inhibitor, LY294002, previously demonstrated to be a pharmacological stimulus for sperm motility. Bicarbonate addition to spermatozoa results in a significant increase in sperm motility as well as in several hyperactivation parameters. This stimulatory effect of bicarbonate and LY294002 is mediated by an increase in cAMP production and tyrosine phosphorylation of the A kinase anchoring protein, AKAP3. The specificity of bicarbonate effects was confirmed by inhibition with 4,4'-di isothiocyanostilbene-2,2'-disulfonic acid. We remark that, in human spermatozoa, bicarbonate acts primarily through activation of sAC to stimulate tyrosine phosphorylation of AKAP3 and sperm motility because both effects are blunted by the sAC inhibitor 2OH-estradiol. In conclusion, our data provide the first evidence that bicarbonate stimulates human sperm motility and hyperactivation through activation of sAC and tyrosine phosphorylation of AKAP3, finally leading to an increased recruitment of PKA to AKAP3. PMID- 15342356 TI - Anti-hyaluronidase oligosaccharide derived from chondroitin sulfate a effectively reduces polyspermy during in vitro fertilization of porcine oocytes. AB - The present study was conducted to examine the effects of chondroitin sulfate A derived oligosaccharide (ChSAO) on hyaluronidase activity and in vitro fertilization (IVF) parameters. The activity of hyaluronidase extracted from preincubated boar sperm was completely blocked by ChSAO at concentrations of 10 microg/ml or higher. After in vitro maturation of porcine cumulus-oocyte complexes, some oocytes were freed from their cumulus cells, and cumulus-intact or cumulus-free oocytes were inseminated with sperm in IVF medium containing various concentrations of ChSAO (0.1-100 microg/ml). In cumulus-intact oocytes, the penetration and the polyspermy rates (39% and 28%, respectively) were significantly decreased by treatment with 100 microg/ml ChSAO compared with those of oocytes treated without ChSAO (63% and 52%, respectively). On the contrary, in cumulus-free oocytes, the addition of 10-100 microg/ml ChSAO significantly reduced the polyspermy rate compared with the control (25-30% versus 53%, respectively), whereas ChSAO had no effect on sperm penetration. Interestingly, ChSAO added to IVF medium significantly decreased the number of sperm bound to the zona pellucida (ZP) of cumulus-free oocytes in a concentration-dependent manner between 0.1 and 100 microg/ml. However, ChSAO had no effect on the time course change in ZP modification after oocyte activation by electrostimulation and the incidence of the acrosome-reacted sperm. Treatment with 100 microg/ml ChSAO during IVF of cumulus-free oocytes significantly increased the proportion of development to the blastocyst stage after in vitro insemination. Therefore, the present findings indicate that hyaluronidase-inhibitory ChSAO is an efficient probe for promoting normal fertilization process in terms of an effective decrease in the incidence of polyspermy during IVF of porcine oocytes. PMID- 15342357 TI - Localization of mitotic arrest deficient 1 (MAD1) in mouse oocytes during the first meiosis and its functions as a spindle checkpoint protein. AB - The present study was designed to investigate the localization of mitotic arrest deficient 1 (MAD1) in mouse oocytes during meiotic maturation and its relationship with kinetochores, chromosomes, and microtubules. Oocytes at various stages during the first meiosis were fixed and immunostained for MAD1, kinetochores, microtubules, and chromosomes. The stained oocytes were examined by confocal microscopy. Some oocytes were treated with nocodazole or Taxol before examination. The anti-MAD1 antibody was injected into the oocytes at the germinal vesicle (GV) stage for examination of chromosome alignment and spindle formation. It was found that MAD1 was present in the oocytes from the GV to prometaphase I stages around the nuclei. When the oocytes reached the metaphase I (M-I) to metaphase II (M-II) stages, MAD1 was mainly localized at the spindle poles. However, MAD1 relocated to the vicinity of the chromosomes when spindles were disassembled by nocodazole or cooling, and the relocated MAD1 moved back to the spindle poles during spindle recovery. Taxol treatment did not affect the MAD1 localization. Although anti-MAD1 antibody injection did not affect nuclear maturation, significantly higher proportions of injected oocytes had misaligned chromosomes when the oocytes reached the M-I to M-II stages. The results of the present study indicate that MAD1 is present in mouse oocytes at all stages during the first meiosis and that it participates in spindle checkpoint during meiosis. However, MAD1 could not check misaligned chromosomes during spindle recovery after the spindles were destroyed by drug or cooling, which caused some chromosomes to scatter in the oocytes. PMID- 15342358 TI - GABAergic integration of progesterone and androgen feedback to gonadotropin releasing hormone neurons. AB - Steroid feedback regulates GnRH secretion and previous work has implicated gamma aminobutyric acid (GABA)ergic neurons as a mediator of these effects. We examined GABAergic postsynaptic currents (PSCs) in green fluorescent protein-identified GnRH neurons from mice exposed to different steroid milieus in vivo. Adult mice were ovariectomized and treated with estradiol (OVX+E, controls) or E plus progesterone (P, OVX+E+P). P decreased PSC frequency, a presynaptic effect, and PSC size, which could be via pre- and/or postsynaptic mechanisms. In contrast, dihydrotestosterone (DHT, OVX+E+DHT) increased both GABAergic PSC frequency and size in GnRH neurons. Tetrodotoxin (TTX), which eliminates action-potential dependent presynaptic effects, did not alter frequency, suggesting DHT may have increased PSC frequency by increasing connectivity between GABAergic and GnRH neurons. TTX reduced PSC size below control values, indicating DHT may augment presynaptic GABA release but inhibits the postsynaptic GnRH neuron response. In mice treated with both P and DHT (OVX+E+P+DHT), PSC frequency and size were similar to controls, suggesting these steroids counteract one another. These results demonstrate GABAergic neurons participate in integrating and conveying steroid feedback to GnRH neurons, defining a potential central mechanism for steroid regulation of GnRH neurons during the reproductive cycle, and providing one possible mechanism for increased activity of these cells in hyperandrogenic females. PMID- 15342359 TI - Validation of an ultrasensitive and specific immunofluorometric assay for mouse follicle-stimulating hormone. AB - Sensitive and specific measurement of FSH is critical to research in reproductive biology, and the increasing availability of transgenic mouse models has created a need for a robust, sensitive, and specific mouse (m) FSH assay. The present study evaluated a time-resolved immunofluorometric assay (IFMA) for mFSH using monoclonal antibody to human (h) FSHbeta as a capture antibody and a biotinylated polyclonal antibody to rat alpha subunit as a detection probe, with signaling amplified by europium-labeled streptavidin. The mFSH IFMA lowered the detection limit 34-fold (5 vs. 170 pg/sample) compared with standard mFSH RIA. The mFSH IFMA demonstrated parallelism of response to dilutions of castrated mouse serum and rat FSH but no cross-reactivity with hFSH and mLH or hLH, whereas the RIA demonstrated nonparallel cross-reactivity with hFSH. The IFMA has a wide analytical range, with a good precision profile for within- and between-assay reproducibility. Because the IFMA is a sandwich-type assay with strict dimer specificity by design, the lower readings and recovery obtained were compared with the RIA when both assays used a pituitary-purified mFSH assay standard that contained isolated or fragmented subunits as well as intact dimeric FSH. When used with mouse serum sample, the mFSH IFMA demonstrated the expected increases following orchidectomy as well as markedly enhanced sensitivity to very low levels of endogenous mFSH in gonadotropin-deficient mice. Furthermore, the IFMA measured mFSH with fidelity in both intact and orchidectomized male mice without any interference from transgenic hFSH. The greatly enhanced sensitivity, specificity, and technical convenience of this mFSH IFMA will allow wider application of FSH measurements to very small blood samples in immature and mature mice as well as transgenic models. PMID- 15342360 TI - Role of motoneurons in the generation of muscle spasms after spinal cord injury. AB - Motoneurons in the spinal cord have intrinsic voltage-dependent persistent inward currents (PICs; e.g. persistent calcium currents) that amplify synaptic inputs by three- to five-fold in addition to providing a sustained excitatory drive that allows motoneurons to fire repetitively following a brief synaptic excitation. In this study, we examined whether prolonged involuntary muscle spasms in subjects with long-term injury to the spinal cord are mediated by the activation of PICs in the motoneuron. To examine this in the human, we used a paired motor unit analysis technique where the firing frequency of one motor unit of the pair (control unit) was used to estimate the synaptic drive to the motoneuron pool, including the drive to a second higher-threshold motor unit of the pair (test unit). The degree to which a motoneuron PIC helped to sustain the discharge of a test motor unit (self-sustained firing) was determined from the reduction in control unit firing at de-recruitment (DeltaF) compared with recruitment of the test unit. This DeltaF value corresponds to the reduction in synaptic drive needed to counteract the intrinsic PIC and, thus, was used an indirect measure of this current. In the nine motor unit pairs studied, the average estimated synaptic drive, or control unit firing rate, required to recruit a test motor unit at the onset of a muscle spasm was significantly higher (by 43%) than the estimated synaptic drive during de-recruitment at the end of a muscle spasm. This indicated that a motoneuron PIC, and associated self-sustained firing, facilitated the firing of the test units during the prolonged muscle spasms. In addition, in all subjects tested (seven out of seven), we observed that following a muscle spasm or voluntary contraction, spontaneous and self-sustained firing of motor units could continue for many seconds, even minutes, at very low discharge rates (average 5.2 +/- 1.6 Hz) with extremely low spike-to-spike variability (coefficient of variation = 5.4 +/- 1.6%). Moreover, increases in synaptic drive (noise) to the spontaneously firing units with voluntary muscle contractions or muscle spasms increased both the mean firing rate of the motor units in addition to their firing variability. This suggests that the slow spontaneous firing commonly observed in chronic spinal injury likely occurs without appreciable synaptic noise and is likely driven to a substantial degree by PICs intrinsic to the motoneuron because it is self-sustained and very regular. PMID- 15342361 TI - Leber hereditary optic neuropathy mtDNA mutations disrupt glutamate transport in cybrid cell lines. AB - Leber hereditary optic neuropathy (LHON) is a maternally inherited form of retinal ganglion cell degeneration leading to optic atrophy which is caused by point mutations in the mitochondrial genome (mtDNA). Three pathogenic mutations (positions 11778/ND4, 3460/ND1 and 14484/ND6) account for the majority of LHON cases and they affect genes that encode for different subunits of mitochondrial complex I. Excitotoxic injury to retinal ganglion cells and the optic nerve has been previously hypothesized, especially given the high susceptibility of this neural cell type to glutamate toxicity. Osteosarcoma-derived cytoplasmic hybrids (cybrids) generated from six unrelated LHON patients, two cell lines for each pathogenic mutation, were compared with cybrids obtained from three healthy controls. Molecular and biochemical analyses showed that excitatory amino acid transporter 1 (EAAT1)/GLAST is the most active glutamate transporter in this cellular model. The glutamate uptake maximal velocity was significantly reduced in all LHON cybrids compared with control cybrids. This reduction was correlated in a mutation-specific fashion with the degree of mitochondrial production of reactive oxygen species, which is enhanced in LHON cybrids. Our findings support the hypothesis that the genetically determined mitochondrial dysfunction in LHON patients leads to impaired activity of the EAAT1 glutamate transporter. This observation is particularly relevant since EAAT1 is the major means of glutamate removal in the inner retina and this prevents retinal ganglion cells being damaged as a result of excitotoxicity. PMID- 15342362 TI - The syndrome of fixed dystonia: an evaluation of 103 patients. AB - We describe the clinical features of 103 patients presenting with fixed dystonia and report the prospective assessment and investigation of 41 of them. Most patients were female (84%) and had a young age of onset [mean 29.7 (SD 13.1) years]. A peripheral injury preceded onset in 63% and spread of dystonia to other body regions occurred in 56%. After an average follow-up of 3.3 years (overall disease duration 8.6 years), partial (19%) or complete (8%) remission had occurred in a minority of patients. The fixed postures affected predominantly the limbs (90%), and rarely the neck/shoulder region (6%) or jaw (4%). In the prospectively studied group, pain was present in most patients and was a major complaint in 41%. Twenty percent of patients fulfilled criteria for Complex Regional Pain Syndrome (CRPS). No consistent investigational abnormalities were found and no patient tested (n = 25) had a mutation in the DYT1 gene. Thirty seven percent of patients fulfilled classification criteria for documented or clinically established psychogenic dystonia; 29% fulfilled DSM-IV (Diagnostic and statistical manual of mental disorders, 4th edition) criteria for somatization disorder, which was diagnosed only after examination of the primary care records in many cases; and 24% fulfilled both sets of criteria. Ten percent of the prospectively studied and 45% of the retrospectively studied patients did not have any evidence of psychogenic dystonia, and detailed investigation failed to reveal an alternative explanation for their clinical presentation. Detailed, semi structured neuropsychiatric assessments in a subgroup of 26 patients with fixed dystonia and in a control group of 20 patients with classical dystonia revealed dissociative (42 versus 0%, P = 0.001) and affective disorders (85 versus 50%, P = 0.01) significantly more commonly in the fixed dystonia group. Medical and surgical treatment was largely unsuccessful. However, seven patients who underwent multidisciplinary treatment, including physiotherapy and psychotherapy, experienced partial or complete remission. We conclude that fixed dystonia usually, but not always, occurs after a peripheral injury and overlaps with CRPS. Investigations are typically normal, but many patients fulfil strict criteria for a somatoform disorder/psychogenic dystonia. In a proportion of patients, however, no conclusive features of somatoform disorder or psychogenic disorder can be found and, in these patients, whether this disorder is primarily neurological or psychiatric remains an open question. Whilst the prognosis is overall poor, remissions do occur, particularly in those patients who are willing and able to undergo multidisciplinary treatment including physiotherapy and psychotherapy, suggesting that this type of treatment should be recommended to these patients. PMID- 15342363 TI - A serial MRI study following optic nerve mean area in acute optic neuritis. AB - This study assessed optic nerve mean area on serial MRI in a cohort of patients with a first episode of acute unilateral optic neuritis to assess the effects of a single acute inflammatory demyelinating lesion. Twenty-nine patients with a median delay from onset of visual symptoms of 13 days (range 7-24 days) were recruited. After a clinical examination and visual evoked potential (VEP) measurement, each patient had their optic nerves imaged with a coronal fat saturated short echo fast fluid-attenuated inversion recovery sequence. Twenty one patients had serial examinations after 2, 4, 8, 12, 26 and 52 weeks. In addition, 32 control subjects had their optic nerves imaged up to three times. The mean cross-sectional area of the intra-orbital portion of each optic nerve was calculated by a blinded observer using a computer-assisted contouring technique. At baseline, the mean area of diseased optic nerves was 16.1 mm2 compared with 13.4 mm2 for healthy contralateral optic nerves (20.1% higher, P < 0.0001) and 13.6 mm2 for controls (18.4% higher, P = 0.0003). The diseased optic nerve mean area declined over time, from initial swelling to later atrophy. The mean decline at 52 weeks was -0.0018 mm2/day (95% confidence interval -0.0038 to 0.00051). At 52 weeks, the mean area of diseased optic nerves was 11.3 mm2 compared with 12.8 mm2 for healthy contralateral optic nerves (11.7% lower, P = 0.032) and 13.1 mm2 for controls (13.7% lower, P = 0.008). The 52 week diseased optic nerve mean area was not significantly affected by the baseline mean area. There was an association between baseline optic nerve mean area and logMAR visual acuity (rS = 0.46, P = 0.012) and visual field mean deviation (rS = -0.55, P = 0.002), but there was no evidence of an association between 1 year mean area and visual outcome. There was no evidence of association between baseline, rates of decline or 1 year diseased optic nerve mean areas and any of the baseline, 1 year or time-averaged VEP variables. The present study shows a consistent pattern of changes associated with individual inflammatory demyelinating lesions in the optic nerve. Acutely, there was swelling, consistent with the presence of acute inflammation, which was related to visual impairment. Over the longer term, there was loss of tissue. The lack of association between 1 year optic nerve mean area and visual outcome may reflect a mild loss of tissue, redundancy or remodelling of function. PMID- 15342364 TI - Increased resistance to activated protein C in women taking third-generation oral contraceptives? PMID- 15342365 TI - A91V is a polymorphism in the perforin gene not causative of an FHLH phenotype. PMID- 15342366 TI - A missense mutation in KIT kinase domain 1 correlates with imatinib resistance in gastrointestinal stromal tumors. AB - KIT gain of function mutations play an important role in the pathogenesis of gastrointestinal stromal tumors (GISTs). Imatinib is a selective tyrosine kinase inhibitor of ABL, platelet-derived growth factor receptor (PDGFR), and KIT and represents a new paradigm of targeted therapy against GISTs. Here we report for the first time that, after imatinib treatment, an additional specific and novel KIT mutation occurs in GISTs as they develop resistance to the drug. We studied 12 GIST patients with initial near-complete response to imatinib. Seven harbored mutations in KIT exon 11, and 5 harbored mutations in exon 9. Within 31 months, six imatinib-resistant rapidly progressive peritoneal implants (metastatic foci) developed in five patients. Quiescent residual GISTs persisted in seven patients. All six rapidly progressive imatinib-resistant implants from five patients show an identical novel KIT missense mutation, 1982T-->C, that resulted in Val654Ala in KIT tyrosine kinase domain 1. This novel mutation has never been reported before, is not present in pre-imatinib or post-imatinib residual quiescent GISTs, and is strongly correlated with imatinib resistance. Allelic-specific sequencing data show that this new mutation occurs in the allele that harbors original activation mutation of KIT. PMID- 15342367 TI - Differential transplantability of tumor-associated stromal cells. AB - At the time of transplantation, tumor fragments contain "passenger" cells: endothelial cells and other stromal cells from the original host. Here, we investigated the fate of genetically labeled endothelial and nonendothelial stromal cells after transplantation in syngeneic mice. We report that angiogenic stroma associated with tumor or adipose tissue persists when transplanted, remains functional, and governs the initial neovascularization of grafted tissue fragments for more than 4 weeks after implantation. Surprisingly, the passenger endothelial cells survive longer than other stromal cells, which are replaced by host-activated fibroblasts after 3 weeks. The transplantability of tumor stroma suggests that the angiogenic potential of a tumor xenograft, which determines its viability, depends on the presence of passenger endothelial cells and other stromal cells within the xenograft. These studies of tumor tissue transplantation provide a platform for exploring the role of epithelial-stromal interactions in studies of tumor heterogeneity and drug resistance. PMID- 15342368 TI - Deficiency of NRH:quinone oxidoreductase 2 increases susceptibility to 7,12 dimethylbenz(a)anthracene and benzo(a)pyrene-induced skin carcinogenesis. AB - NRH:Quinone oxidoreductase 2 (NQO2) is an enzyme that catalyzes the reductive metabolism of quinones. C57BL/6 NQO2-/- mice lacking NQO2 gene expression were generated in our laboratory. The dorsal skin of NQO2-deficient mice was exposed to 7,12-dimethylbenz(a)anthracene (DMBA) or benzo(a)pyrene alone (complete carcinogen) or with 12-O-tetradecanoylphorbol-13-acetate (TPA) (initiation/promotion model) to determine the in vivo role of NQO2 in chemical carcinogenesis. The NQO2-/- mice showed significantly increased tumor frequency with DMBA + TPA when compared with their wild-type littermates. The benzo(a)pyrene + TPA also showed increase in tumor incidence in NQO2-/- mice but to a less extent than DMBA. DMBA alone resulted in low frequency of tumor development with no difference in susceptibility between wild-type and NQO2-/- mice. Benzo(a)pyrene alone failed to induce tumors in either wild-type or NQO2-/- mice. Histologic analysis of the NQO2-/- mice tumors demonstrated proliferative activity. The treatment of NQO2-/- mice skin with benzo(a)pyrene failed to significantly increase tumor suppressor protein p53 and p53-regulated growth related protein p21 and proapoptotic protein Bax as observed in case of wild-type mice. These results demonstrate that NQO2 protects against DMBA- and benzo(a)pyrene-induced skin carcinogenesis and suggest that NQO2 protection might be against tumor promotion. The results also suggest that lack of induction of p53, p21, and Bax proteins might contribute to increased sensitivity of NQO2-/- mice skin to benzo(a)pyrene carcinogenicity. PMID- 15342369 TI - Protein profiling of microdissected prostate tissue links growth differentiation factor 15 to prostate carcinogenesis. AB - Identification of proteomic alterations associated with early stages in the development of prostate cancer may facilitate understanding of progression of this highly variable disease. Matched normal, high-grade prostatic intraepithelial neoplasia (hPIN) and prostate cancer cells of predominantly Gleason grade 3 were procured by laser capture microdissection from serial sections obtained from snap-frozen samples dissected from 22 radical prostatectomy specimens. From these cells, protein profiles were generated by surface-enhanced laser desorption/ionization-time of flight mass spectrometry. A 24-kDa peak was observed at low or high intensity in profiles of prostate cancer cells in 19 of 27 lesions and at low intensity in 3 of 8 hPIN lesions but was not detectable in matched normal cells. SDS-PAGE analysis of prostate cancer and matched normal epithelium confirmed expression of a prostate cancer-specific 24 kDa protein. Mass spectrometry and protein data-based analysis identified the protein as the dimeric form of mature growth differentiation factor 15 (GDF15). The increased expression of mature GDF15 protein in prostate cancer cells cannot be explained on the basis of up-regulation of GDF15 mRNA because reverse transcription-PCR analysis showed similar amounts of transcript in normal, hPIN, and prostate cancer cells that were obtained by laser capture microdissection in the same set of serial sections from which the protein profiles were obtained. Our findings suggest that early prostate carcinogenesis is associated with expression of mature GDF15 protein. PMID- 15342370 TI - alpha-type-1 polarized dendritic cells: a novel immunization tool with optimized CTL-inducing activity. AB - Using the principle of functional polarization of dendritic cells (DCs), we have developed a novel protocol to generate human DCs combining the three features critical for the induction of type-1 immunity: (a) fully mature status; (b) responsiveness to secondary lymphoid organ chemokines; and (c) high interleukin 12p70 (IL-12p70)-producing ability. We show that IFN-alpha and polyinosinic:polycytidylic acid (p-I:C) synergize with the "classical" type-1 polarizing cytokine cocktail [tumor necrosis factor alpha (TNFalpha)/IL 1beta/IFNgamma], allowing for serum-free generation of fully mature type-1 polarized DCs (DC1). Such "alpha-type-1-polarized DC(s)" (alphaDC1) show high migratory responses to the CCR7 ligand, 6C-kine but produce much higher levels of IL-12p70 as compared to TNFalpha/IL-1beta/IL-6/prostaglandin E2 (PGE2)-matured DCs (sDC), the current "gold standard" in DC-based cancer vaccination. A single round of in vitro sensitization with alphaDC1 (versus sDCs) induces up to 40-fold higher numbers of long-lived CTLs against melanoma-associated antigens: MART-1, gp100, and tyrosinase. Serum-free generation of alphaDC1 allows, for the first time, the clinical application of DCs that combine the key three features important for their efficacy as anticancer vaccines. PMID- 15342371 TI - A new role of protein phosphatase 2a in adenoviral E1A protein-mediated sensitization to anticancer drug-induced apoptosis in human breast cancer cells. AB - The adenoviral type 5 E1A protein has been shown to induce sensitization to different categories of anticancer drug-induced apoptosis, partly by down regulation of the activity of a critical oncogenic kinase Akt in both normal fibroblasts and epithelial breast cancer cells. Currently, the adenoviral E1A gene is being tested as an antitumor gene in multiple clinical trials. However, molecular mechanisms underlying E1A-mediated chemosensitization and down regulation of Akt activity are still not completely defined. Here, we show that E1A by up-regulation of the catalytic subunit of protein phosphatase 2A [PP2A (PP2A/C)] enhanced the activity of PP2A, which results in repression of Akt activation in E1A-expressing cells. In addition, activation of PP2A/C is required for E1A-mediated sensitization to drug-induced apoptosis, because blocking PP2A/C expression using a specific small interfering RNA against PP2A/C reduced drug sensitivity in E1A-expressing cells. Deletion mutation of the conserved domain of E1A, which is required for E1A-mediated sensitization to drug-induced apoptosis, also abolished the ability of E1A to up-regulate PP2A/C. Thus, the up-regulation of PP2A may represent a novel mechanism for E1A-mediated sensitization to anticancer drug-induced apoptosis. PMID- 15342372 TI - XBP1 is essential for survival under hypoxic conditions and is required for tumor growth. AB - Hypoxia within solid tumors is a major determinant of outcome after anticancer therapy. Analysis of gene expression changes during hypoxia indicated that unfolded protein response genes were one of the most robustly induced groups of genes. In this study, we investigated the hypoxic regulation of X-box binding protein (XBP1), a major transcriptional regulator of the unfolded protein response. Hypoxia induced XBP1 at the transcriptional level and activated splicing of its mRNA, resulting in increased levels of activated XBP1 protein. After exposure to hypoxia, apoptosis increased and clonogenic survival decreased in XBP1-deficient cells. Loss of XBP1 severely inhibited tumor growth due to a reduced capacity for these transplanted tumor cells to survive in a hypoxic microenvironment. Taken together, these studies directly implicate XBP1 as an essential survival factor for hypoxic stress and tumor growth. PMID- 15342373 TI - Genome-wide array-based comparative genomic hybridization of diffuse large B-cell lymphoma: comparison between CD5-positive and CD5-negative cases. AB - Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin's lymphoma and exhibits aggressive and heterogeneous clinical behavior. To genetically characterize DLBCL, we established our own array-based comparative genomic hybridization and analyzed a total of 70 cases [26 CD-positive (CD5+) DLBCL and 44 CD5-negative (CD5-) DLBCL cases]. Regions of genomic aberrations observed in >20% of cases of both the CD5+ and CD5- groups were gains of 1q21 q31, 1q32, 3p25-q29, 5p13, 6p21-p25, 7p22-q31, 8q24, 11q23-q24, 12q13-q21, 16p13, 18, and X and losses of 1p36, 3p14, 6q14-q25, 6q27, 9p21, and 17p11-p13. Because CD5 expression marks a subgroup with poor prognosis, we subsequently analyzed genomic gains and losses of CD5+ DLBCL compared with those of CD5-. Although both groups showed similar genomic patterns of gains and losses, gains of 10p14-p15 and 19q13 and losses of 1q43-q44 and 8p23 were found to be characteristic of CD5+ DLBCL. By focusing on the gain of 13q21-q34 and loss of 1p34-p36, we were also able to identify prognostically distinct subgroups among CD5+ DLBCL cases. These results suggest that array-based comparative genomic hybridization analysis provides a platform of genomic aberrations of DLBCL both common and specific to clinically distinct subgroups. PMID- 15342374 TI - CpG hypermethylation of MDR1 gene contributes to the pathogenesis and progression of human prostate cancer. AB - Multidrug resistance 1 (MDR1) gene encodes for P-glycoprotein (P-gp), a Mr 170,000 transmembrane calcium-dependent efflux pump that is inactivated in prostate cancer. We hypothesize that inactivation of the MDR1 gene through CpG methylation contributes to the pathogenesis and progression of prostate cancer. To test this hypothesis, CpG methylation status of the MDR1 promoter and its correlation with clinicopathological findings were evaluated in 177 prostate cancer samples and 69 benign prostate hypertrophy (BPH) samples. Cellular proliferation index and apoptotic index were determined by proliferating cell nuclear antigen (PCNA) and single-strand DNA immunostaining, respectively. After 5-aza-2'-deoxycytidine treatment, increased expression of MDR1 mRNA transcript was found in prostate cancer cell lines (DU145, DuPro, and ND1). MDR1 methylation frequency was significantly higher in prostate cancer samples compared with BPH samples (54.8 versus 11.6%, respectively, P < 0.001). Logistic regression analysis revealed that PC patients are 11.5 times more likely to have MDR1 methylation than BPH patients (95% confidence interval 4.87-27.0) and that MDR1 methylation is independent of the age. Significant correlation of MDR1 methylation was observed with high pT category (P < 0.001), high Gleason sum (P = 0.008), high preoperative prostate-specific antigen (P = 0.01), and advancing pathological features. In addition, PCNA-labeling index were significantly higher in methylation-specific PCR (MSP)-positive than in MSP-negative prostate cancer samples (P = 0.048). In contrast, no significant difference in apoptotic index was found between MSP-positive and -negative prostate cancer samples. These findings suggest that CpG hypermethylation of MDR1 promoter is a frequent event in prostate cancer and is related to disease progression via increased cell proliferation in prostate cancer cells. PMID- 15342375 TI - Molecular features of the transition from prostatic intraepithelial neoplasia (PIN) to prostate cancer: genome-wide gene-expression profiles of prostate cancers and PINs. AB - To characterize the molecular feature in prostate carcinogenesis and the putative transition from prostatic intraepithelial neoplasia (PIN) to invasive prostate cancer (PC), we analyzed gene-expression profiles of 20 PCs and 10 high-grade PINs with a cDNA microarray representing 23,040 genes. Considering the histological heterogeneity of PCs and the minimal nature of PIN lesions, we applied laser microbeam microdissection to purify populations of PC and PIN cells, and then compared their expression profiles with those of corresponding normal prostatic epithelium also purified by laser microbeam microdissection. A hierarchical clustering analysis separated the PC group from the PIN group, except for three tumors that were morphologically defined as one very-high-grade PIN and two low-grade PCs, suggesting that PINs and PCs share some molecular features and supporting the hypothesis of PIN-to-PC transition. On the basis of this hypothesis, we identified 21 up-regulated genes and 63 down-regulated genes commonly in PINs and PCs compared with normal epithelium, which were considered to be involved in the presumably early stage of prostatic carcinogenesis. They included AMACR, OR51E2, RODH, and SMS. Furthermore, we identified 41 up-regulated genes and 98 down-regulated genes in the transition from PINs to PCs; those altered genes, such as POV1, CDKN2C, EPHA4, APOD, FASN, ITGB2, LAMB2, PLAU, and TIMP1, included elements that are likely to be involved in cell adhesion or the motility of invasive PC cells. The down-regulation of EPHA4 by small interfering RNA in PC cells lead to attenuation of PC cell viability. These data provide clues to the molecular mechanisms underlying prostatic carcinogenesis, and suggest candidate genes the products of which might serve as molecular targets for the prevention and treatment of PC. PMID- 15342376 TI - Global expression profiling identifies signatures of tumor virulence in MMTV-PyMT transgenic mice: correlation to human disease. AB - FVB/N-Tg (MMTV-PyMT)(634Mul)-transgenic mice develop multifocal mammary tumors with a high incidence of pulmonary metastasis. We have demonstrated previously that mammary tumors derived from transgene-positive F1 progeny in particular inbred strains display altered latency, tumor growth rates, and metastatic rates when compared with the FVB/NJ homozygous parent. To identify genes with expression that might be critical in modifying the biological behavior of MMTV PyMT tumors, we performed a detailed comparative analysis of expression profiles from mammary tumors arising in the parental FVB/NJ background and F1 progeny from crosses with I/LnJ, LP/J, MOLF/Ei, and NZB/B1NJ mice. Compared with normal mammary glands, gene expression profiles of tumors from all five strains exhibited up-regulation of genes involved in cell growth (e.g., Cks1 and CDC25C) and down-regulation of cell adhesion molecules, with many genes associated previously with human breast cancer such as STAT2, CD24 antigen, gelsolin, and lipocalin2. To identify genes with significant variation in expression between the five different genotypes, significance analysis of microarrays (SAM) and one way ANOVA were used. Three definable groupings of tumors were identified: (a) tumors derived in the LP/J F1 and MOLF/Ei F1 strains in which tumor growth and dissemination are suppressed and latency prolonged; (b) the most aggressive tumors from the FVB/NJ parental strain and I/LnJ F1 genomic backgrounds; and (c) an intermediate virulence phenotype with tumors from NZB/B1NJ-F1 crosses. These array based assessments correlated well with a composite phenotype ranking using a "virulence" index. The gene expression signature that is associated with a high metastatic rate in the mouse contains the same 17 genes described recently as the signature gene set predictive of metastasis in human tumors (1) with 16 of the 17 genes exhibiting the same directional change in expression associated with human metastases. These results demonstrate that the genetic analysis of mouse models of tumorigenesis may be highly relevant to human cancer and that the metastatic phenotype of a tumor may be affected by the germline genetic configuration of the host. PMID- 15342377 TI - Inverse correlation between cyclin A1 hypermethylation and p53 mutation in head and neck cancer identified by reversal of epigenetic silencing. AB - Aberrant promoter hypermethylation of tumor suppressor genes is proposed to be a common feature of primary cancer cells. We recently developed a pharmacological unmasking microarray approach to screen unknown tumor suppressor gene candidates epigenetically silenced in human cancers. In this study, we applied this method to identify such genes in head and neck squamous cell carcinoma (HNSCC). We identified 12 novel methylated genes in HNSCC cell lines, including PGP9.5, cyclin A1, G0S2, bone-morphogenetic protein 2A, MT1G, and neuromedin U, which showed frequent promoter hypermethylation in primary HNSCC (60%, 45%, 35%, 25%, 25%, and 20%, respectively). Moreover, we discovered that cyclin A1 methylation was inversely related to p53 mutational status in primary tumors (P = 0.015), and forced expression of cyclin A1 resulted in robust induction of wild-type p53 in HNSCC cell lines. Pharmacological unmasking followed by microarray analysis is a powerful tool to identify key methylated tumor suppressor genes and relevant pathways. PMID- 15342378 TI - Frequent nonrandom activation of germ-line genes in human cancer. AB - The growing class of cancer/germ-line genes is characterized by a unique expression pattern with transcription restricted to germ cells and cancer cells. It is not known which fraction of germ-line genes is ectopically activated in tumor cells and whether this fraction displays common features as compared with strictly germ-line genes remaining silent in cancer. Using an unbiased genome wide scanning approach, representative samples of both cancer/germ-line genes as well as strictly germ-line-specific genes were determined. Comparative analysis disclosed highly significant diametric characteristics for these two categories of genes with regard to sex specificity, developmental stage of physiological expression during gametogenesis, chromosomal localization, and epigenetic regulation of expression. Our findings provide class predictors for germ cell specific gene activation in cancer. The identification of highly congruent expression patterns in cancer and in DNA methyltransferase-deficient cells suggests an underlying common epigenetic mechanism for activation of germ-line genes in cancer. PMID- 15342379 TI - 1,1-Bis(3'-indolyl)-1-(p-substitutedphenyl)methanes induce peroxisome proliferator-activated receptor gamma-mediated growth inhibition, transactivation, and differentiation markers in colon cancer cells. AB - 1,1-Bis(3'indolyl)-1-(p-substitutedphenyl)methanes containing p-trifluoromethyl (DIM-C-pPhCF)), p-t-butyl (DIM-C-pPhtBu), and p-phenyl (DIM-C-pPhC6H5) groups induce peroxisome proliferator-activated receptor gamma (PPARgamma)-mediated transactivation in HT-29, HCT-15, RKO, and SW480 colon cancer cell lines. Rosiglitazone also induces transactivation in these cell lines and inhibited growth of HT-29 cells, which express wild-type PPARgamma but not HCT-15 cells, which express mutant (K422Q) PPARgamma. In contrast, DIM-C-pPhCF3, DIM-C-pPhtBu, and DIM-C-pPhC6H5 inhibited growth of both HT-29 and HCT-15 cells with IC50 values ranging from 1 to 10 micromol/L. Rosiglitazone and diindolylmethane (DIM) analogues did not affect expression of cyclin D1, p21, or p27 protein levels or apoptosis in HCT-15 or HT-29 cells but induced keratin 18 in both cell lines. However, rosiglitazone induced caveolins 1 and 2 in HT-29 but not HCT-15 cells, whereas these differentiation markers were induced by DIM-C-pPhCF3 and DIM-C pPhC6H5 in both cell lines. Because overexpression of caveolin 1 is known to suppress colon cancer cell and tumor growth, the growth inhibitory effects of rosiglitazone and the DIM compounds are associated with PPARgamma-dependent induction of caveolins. PMID- 15342380 TI - Identification of aberrantly methylated genes in association with adult T-cell leukemia. AB - In this study, we identified 53 aberrantly hypermethylated DNA sequences in adult T-cell leukemia (ATL) cells using methylated CpG island amplification/representational difference analysis method. We also observed a proportionate increase in the methylation density of these regions with disease progression. Seven genes, which were expressed in normal T cells, but suppressed in ATL cells, were identified near the hypermethylated regions. Among these silenced genes, Kruppel-like factor 4 (KLF4) gene is a cell cycle regulator and early growth response 3 (EGR3) gene is a critical transcriptional factor for induction of Fas ligand (FasL) expression. Treatment with 5-aza-2'-deoxycytidine resulted in the recovery of their transcription, indicating that their silencing might be associated with DNA hypermethylation. To study their functions in ATL cells, we transfected recombinant adenovirus vectors expressing KLF4 and EGR3 genes. Expression of KLF4 induced apoptosis of ATL cells whereas enforced expression of EGR3 induced the expression of FasL gene, resulting in apoptosis. Thus, suppressed expression of EGR3 enabled ATL cells to escape from activation induced cell death mediated by FasL. Our results showed that the methylated CpG island amplification/representational difference analysis method allowed the isolation of hypermethylated DNA regions specific to leukemic cells and thus shed light on the roles of DNA methylation in leukemogenesis. PMID- 15342381 TI - Tumor-dependent kinetics of partial pressure of oxygen fluctuations during air and oxygen breathing. AB - The primary purpose of this study was to examine the kinetics of partial pressure of oxygen (pO2) fluctuations in fibrosarcoma (FSA) and 9L tumors under air and O2 breathing conditions. The overall hypothesis was that key factors relating to oxygen tension fluctuations would vary between the two tumor types and as a function of the oxygen content of the breathing gas. To assist in the interpretation of the temporal data, spatial pO2 distributions were measured in 10 FSA and 8 9L tumors transplanted into the subcutis of the hind leg of Nembutal anesthetized (50 mg/kg) Fischer 344 rats. Recessed-tip oxygen microelectrodes were inserted into the tumor, and linear pO2 measurements were recorded in 50 microm steps along a 3-mm path, and blood pressure was simultaneously measured via femoral arterial access. Additionally, pO2 was measured at a single location for 90 to 120 minutes in FSA (n=11) or 9L tumors (n=12). Rats were switched from air to 100% O2 breathing after 45 minutes. Temporal pO2 records were evaluated for their potential radiobiological significance by assessing the number of times they crossed a 10-mm-Hg threshold. In addition, the data were subjected to Fourier analysis for air and O2 breathing. FSA and 9L tumors had spatial median pO2 measurements of 4 and 1 mm Hg, respectively. 9L had more low pO2 measurements < or =2.5 mm Hg than did FSA, whereas between 2.5 and 10 mm Hg this pattern was reversed. Pimonidazole staining patterns in FSA and 9L tumors supported these results. Temporal pO2 instability was observed in all experiments during air and O2 breathing. Threshold analyses indicated that the 10 mm Hg threshold was crossed 2 to 5 times per hour, independent of tumor type. However, the magnitude of 9L pO2 fluctuations was approximately eight times greater than FSA fluctuations, as assessed with Fourier transform analysis (Wilcoxon, P < 0.005). O2 breathing significantly increased median pO2 in FSA from 3 to 8 mm Hg (P < 0.005) and caused a significant increase in frequency and magnitude of pO2 fluctuations. One hundred percent O2 breathing had no effect on 9L tumor pO2, and it decreased the magnitude of pO2 fluctuations with borderline significance. These results show that these two tumors differ significantly with respect to spatial and temporal oxygenation conditions under air and O2 breathing. Fluctuations of pO2 of the type reported herein are predicted to significantly affect radiotherapy response and could be a source for genetic instability, increased angiogenesis, and metastases. PMID- 15342382 TI - Conditional deletion of Rb causes early stage prostate cancer. AB - Prostate cancer remains the second leading cause of cancer-related death for men in the United States. Mutations in tumor suppressor genes including retinoblastoma (Rb), p53, and PTEN have been linked to the development of prostate cancer in man and mouse models, and loss of heterozygosity of the Rb locus has been observed in up to 60% of clinical cases. In this study we demonstrate that conditional somatic deletion of even a single Rb allele in the epithelial cells of the mouse prostate causes focal hyperplasia, thereby establishing a causal relationship between Rb loss and development of early stage prostate cancer. As a consequence of Rb ablation we observed increased expression of E2F target genes and a concomitant increase in proliferation in the epithelial compartment. However, by 52 weeks of age these lesions had not become malignant and represent an early stage of the disease. Nevertheless, the multifocal nature of the phenotype in the mice closely resembled multifocality of clinical disease. Taken together, our data demonstrated that loss of pRB-mediated cell cycle control directly caused the initiation of proliferative prostate disease but was insufficient to cause malignancy. Establishment of this early initiation model will aid efforts to thoroughly characterize early prostate disease as well as the elucidation of molecular mechanisms that cooperate with Rb loss to facilitate progression and metastasis. PMID- 15342383 TI - Selective usage of D-Type cyclins by Ewing's tumors and rhabdomyosarcomas. AB - The genetic mechanisms that control proliferation of childhood musculoskeletal malignancies, notably Ewing's tumor (ET) and rhabdomyosarcoma (RMS), remain largely unknown. Most human cancers appear to overexpress at least one of the G1 cyclins (cyclins D1, D2, D3, E1, and E2) to bypass normal regulation of cell cycle G1 progression. We compared the gene expression profiles of 7 ET and 13 RMS primary tumor samples and found overexpression of cyclin D1 in all 7 ET samples. In contrast, RMS samples expressed higher levels of cyclin D2, cyclin D3, and cyclin E1. This was confirmed by quantitative reverse transcription-polymerase chain reaction and Western blot. The relative roles of RAS-extracellular signal regulated kinase 1/2 and phosphatidylinositol 3'-kinase (PI3K)-AKT pathways in the regulation of D-type cyclin expression in these tumors were then assessed. Inhibition of either pathway reduced expression of cyclins D1, D2, and D3 in RMS lines, whereas only PI3K inhibitors blocked cyclin D1, D2, and D3 expression in ET lines. Furthermore, PI3K-AKT appeared to regulate D-type cyclin transcription in RMS lines through FKHR and FKHRL1. Finally, the role of the ET-associated EWS FLI1 fusion gene in regulating D cyclin expression was studied. Inhibition of EWS FLI1 expression in the TC71 ET line decreased cyclin D1 levels but increased cyclin D3 levels. In contrast, induction of EWS-FLI1 expression in the RD RMS cell line increased cyclin D1 expression but decreased cyclin D3 expression. Our results demonstrate distinct regulation of D-type cyclins in ET and RMS and indicate that EWS-FLI1 can modulate the expression of D-type cyclins independent of cellular backgrounds. PMID- 15342384 TI - Mek1 alters epidermal growth and differentiation. AB - The highly homologous kinases, Mek1 and Mek2, act downstream of Ras and Raf to activate extracellular signal-regulated kinase (ERK) mitogen-activated protein kinases. In epidermis, Ras and Raf promote hyperplasia; however, they act on multiple Mek-independent effectors, and the extent to which Meks can mediate these effects is unknown. To address this, we expressed inducible Meks in transgenic murine and human epidermis. Both Mek1 and Mek2 triggered ERK phosphorylation. Only Mek1, however, recapitulated Ras/Raf effects in increasing proliferation and integrin expression while suppressing differentiation, which are impacts characteristic of epidermal neoplasia. Furthermore, a kinase-dead Mek1 mutant incapable of phosphorylating ERK proteins retained ability to mediate Mek1-driven epidermal proliferation. Mek1 is thus sufficient to promote the proliferative epithelial phenotype in a manner independent of intact kinase function. PMID- 15342385 TI - The Rgr oncogene induces tumorigenesis in transgenic mice. AB - To study the oncogenic potential of Rgr in vivo, we have generated several transgenic Rgr mouse lines, which express the oncogene under the control of different promoters. These studies revealed that Rgr expression leads to the generation of various pathological alterations, including fibrosarcomas, when its transgenic expression is restricted to nonlymphoid tissues. Moreover, the overall incidence and latency of fibrosarcomas were substantially increased and shortened, respectively, in a p15INK4b-defective background. More importantly, we also have demonstrated that Rgr expression in thymocytes of transgenic mice induces severe alterations in the development of the thymocytes, which eventually lead to a high incidence of thymic lymphomas. This study demonstrates that oncogenic Rgr can induce expression of p15INK4b and, more importantly, that both Rgr and p15INK4b cooperate in the malignant phenotype in vivo. These findings provide new insights into the tumorigenic role of Rgr as a potent oncogene and show that p15INK4b can act as a tumor suppressor gene. PMID- 15342386 TI - Hath1, down-regulated in colon adenocarcinomas, inhibits proliferation and tumorigenesis of colon cancer cells. AB - A striking feature of colon tumors is the significant reduction of goblet cells. Although targeted deletion of Math1 in mice leads to a loss of intestinal secretory cells, including goblet cells, the role of Hath1 in colon tumorigenesis remains unknown. Here we report that Hath1, the human ortholog of Math1, was dramatically down-regulated in colon tumor samples and colon cancer cell lines. Overexpression of Hath1 in HT29, an aggressive colon cancer cell line, resulted in a significant inhibition on cell proliferation, anchorage-independent growth in soft agar and, more importantly, growth of human colon cancer cell xenografts in athymic nude mice. Such inhibition was accompanied by altered expression of a goblet cell differentiation marker, MUC2, and cell cycle regulators cyclin D1 and p27kip1. Hath1 expression also was up-regulated on inhibition of the Wnt pathway, which has been well implicated in colon tumorigenesis. Hence, this study suggests that Hath1 may be a novel factor downstream of the Wnt pathway capable of suppressing anchorage-independent growth of colon cancer cell lines. More importantly, this study is the first to establish a link between down-regulation of Hath1 expression and colon tumorigenesis. PMID- 15342387 TI - Molecular profiling of human hepatocellular carcinoma defines mutually exclusive interferon regulation and insulin-like growth factor II overexpression. AB - Molecular subtyping of human hepatocellular carcinoma (HCC) with potential mechanistic and therapeutic impact has not been achieved thus far. We have analyzed the mRNA expression patterns of 43 different human HCC samples and 3 HCC cell lines in comparison with normal adult liver using high-density cDNA microarrays. Two main groups of HCC, designated group A (65%) and group B (35%), were distinguished based on clustering of the most highly varying genes. Group A HCCs were characterized by induction of a number of interferon (IFN)-regulated genes, whereas group B was characterized mainly by down-regulation of several apoptosis-relevant and IFN-regulated genes. The number of apoptotic tumor cells and tumor-infiltrating lymphocytes was significantly higher in tumors of group A as compared with those of group B. Based on the expression pattern, group B was further subdivided into two subgroups, designated subgroup B1 (6 of 43 tumors, 14%) and subgroup B2 (9 of 43 tumors, 21%). A prominent characteristic of subgroup B1 was high overexpression of insulin-like growth factor (IGF)-II. All tested HCC cell lines expressed equally high concentrations of IGF-II transcripts and co-segregated with group B1 in clustering. IGF-II overexpression and induction of IFN-related genes were mutually exclusive, even when analysis was extended to other cancer expression profile studies. Moreover, IFN-gamma treatment substantially reduced IGF-II expression in HCC cells. In conclusion, cDNA microarray analyses provided subtyping of HCCs that is related to intratumor inflammation and tumor cell apoptosis. This profiling may be of mechanistic and therapeutic impact because IGF-II overexpression has been linked to reduced apoptosis and increased proliferation and may be accessible to therapeutic intervention. PMID- 15342388 TI - Adhesion of gastric carcinoma cells to peritoneum mediated by alpha3beta1 integrin (VLA-3). AB - The interaction between gastric carcinoma cells and the peritoneal lining is a key step in peritoneal dissemination. In this study, we examined the roles of the beta1 family of integrin receptors in the adhesion of such cells to the peritoneum. The adhesion of several gastric carcinoma cell lines to peritonea excised from mice was inhibited most by an anti-alpha3 integrin antibody and to a lesser extent by an anti-alpha2 integrin antibody. In the peritoneal implantation of NUGC-4 human gastric carcinoma cells in athymic mice, treatment of the cells with anti-alpha2 or anti-alpha3 integrin antibody reduced the number of disseminated nodules; suppression by the anti-alpha3 integrin antibody was stronger than that by the anti-alpha2 integrin antibody. The cDNAs to human alpha2 and alpha3 integrins were introduced into K562 leukemic cells, which were positive for the integrin beta1 subunit but negative for the alpha2 or alpha3 subunit. The alpha3 integrin-transfected cells adhered to excised peritoneum and to a monolayer of peritoneal mesothelial cells more firmly than did the alpha2 integrin-transfected cells or the mock transfectant. Reverse transcription-PCR was used to analyze the expression of laminin-5 and laminin-10/11, which have been reported to serve as high-affinity ligands for alpha3beta1 integrin. mRNA for these laminin isoforms was found in mesothelial cells from the diaphragm and parietal peritoneum. These results strongly suggest that alpha3beta1 integrin plays an essential role in mediating the initial attachment of cancer cells to the peritoneum, leading to the formation of peritoneal metastasis. PMID- 15342389 TI - Hedgehog signaling pathway is a new therapeutic target for patients with breast cancer. AB - The Hedgehog (Hh) signaling pathway functions as an organizer in embryonic development. Genetic analysis has demonstrated a critical role for the Hh pathway in mammary gland morphogenesis. Disruption of Patched1, a component of the Hh pathway, results in abnormal growth of mammary duct. Recent studies have shown constitutive activation of the Hh pathway in various types of malignancies. However, it remains unclear whether this pathway is activated in human breast cancer. Here, we determined the expression of the components, including Sonic Hh, Patched1, and Gli1, of the Hh pathway by immunohistochemical staining in a series of 52 human breast carcinomas. All of 52 tumors display staining of high intensity for Gli1 when compared with adjacent normal tissue. The nuclear staining ratio of Gli1 correlates with expression of estrogen receptor and histologic type. Exposure to cyclopamine, a steroidal alkaloid that blocks the Hh pathway, suppresses expression of Gli1 and the growth of the Hh pathway-activated breast carcinoma cells. These data indicate that the Hh pathway is a new candidate for therapeutic target of breast cancer. PMID- 15342390 TI - CITED4 inhibits hypoxia-activated transcription in cancer cells, and its cytoplasmic location in breast cancer is associated with elevated expression of tumor cell hypoxia-inducible factor 1alpha. AB - The interaction of hypoxia-inducible factor 1alpha and the CH1 domain of the transcriptional coactivator p300/CBP is necessary for the expression of hypoxia responsive genes and tumor angiogenesis. The transcription factor CITED2 binds p300/CBP at the CH1 domain and functions as a negative regulator of hypoxia signaling by competing with hypoxia-inducible factor 1alpha. CITED4, a recently identified member of the CITED family, binds p300/CBP via the CH1 domain and functions as a coactivator for transcription factor AP-2. Here, we show that CITED4 blocks the binding of hypoxia-inducible factor 1alpha to p300 in vitro and inhibits hypoxia-inducible factor-1alpha transactivation and hypoxia-mediated reporter gene activation. These studies suggest that CITED4 might function as an inhibitor of hypoxia-inducible factor 1alpha. To explore the function of CITED4 in breast cancer, we determined its expression in normal, in situ and invasive breast cancers. We also correlated its expression in 286 invasive breast tumors with clinicopathological, hypoxia markers and survival. In contrast to the nuclear localization of CITED4 in normal breast tissue, breast tumors were characterized by cytoplasmic and nuclear localization. Nuclear CITED4 expression was significantly inversely associated with tumor hypoxia-inducible factor 1alpha (P < 0.05), tumor size (P = 0.03), tumor grade (P = 0.0001), and Chalkley vessel count (P = 0.04). CITED4 showed no significant correlation with patient age (P = 0.45), estrogen receptor (P = 0.11), or epidermal growth factor receptor (P = 0.48). These results show that breast cancer development is characterized by either nuclear loss or cytoplasmic translocation of CITED4, with consequent loss of hypoxia-inducible factor-1alpha transcriptional antagonist activity. This may be an important mechanism by which tumors enhance hypoxia-inducible factor expression and result in an aggressive phenotype. PMID- 15342391 TI - Growth and survival mechanisms associated with perineural invasion in prostate cancer. AB - Perineural invasion (PNI) is the major mechanism of prostate cancer spread outside the prostate. Apoptotic and proliferation indices were determined in PNI cells using the PNI in vitro model and human PNI in tissue microarrays. RNA was extracted from the PNI model and controls and evaluated by cDNA microarray analysis. Differential expression of candidate genes was confirmed by real-time quantitative PCR, fluorescence, and immunohistochemistry using tissue microarrays. Genistein and BAY 11-7085 were added to the supernatant of cocultures and controls in microchamber cultures. The significance of nuclear factor kappaB (NFkappaB) nuclear translocation in human PNI was analyzed using Kaplan-Meier analysis. An increase in proliferation and a decrease in apoptosis were observed in human PNI cells and the PNI model as compared with controls. Three of 15 genes up-regulated in the cDNA microarray were involved in the apoptosis signaling pathway (NFkappaB), and its downstream targets defender against cell death 1 and PIM-2. The increase was corroborated by real-time quantitative PCR and immunofluorescence. NFkappaB nuclear translocation was seen in the in vitro model and human tissues, where strong nuclear expression was associated with a decrease in recurrence-free survival. Addition of genistein and BAY 11-7085 resulted in a decrease in NFkappaB, PIM-2 and defender against cell death 1 as well as a reversal of the inhibition of apoptosis. This is the first description of a biological mechanism and functional significance of PNI. Cancer cells in a perineural location acquire a survival and growth advantage using a NFkappaB survival pathway. Targeting PNI might help detain local spread of the tumor and influence survival. PMID- 15342392 TI - TEL2, an ETS factor expressed in human leukemia, regulates monocytic differentiation of U937 Cells and blocks the inhibitory effect of TEL1 on ras induced cellular transformation. AB - TEL2 is a member of the ETS family of transcription factors, which is highly similar to TEL1/ETV6. It binds to DNA via the ETS domain and interacts with itself or TEL1 via the pointed domain. The expression of TEL2 in normal and leukemic hematopoietic cells suggests a role in hematopoietic development. In this article, we describe the role of TEL2 in hematopoietic differentiation and cellular transformation. Quantitative reverse transcription-PCR showed that the expression of TEL2 mRNA was down-regulated during monocytic differentiation of U937 and HL60 induced by 1,25-(OH)2 vitamin D3 and 12-O-tetradecanoylphorbol 13 acetate, respectively. Overexpression of TEL2 in U937 cells inhibited differentiation induced by vitamin D3. In contrast, overexpression of a TEL2 mutant lacking either the pointed domain or a functional ETS domain induced both differentiation of U937 cells and inhibited their growth in vitro and in vivo. In addition, these mutants blocked TEL2-mediated transcriptional repression of a synthetic promoter containing TEL2 binding sites. These data suggest that dominant-negative inhibition of TEL2 might cause differentiation. Quantitative reverse transcription-PCR demonstrated that TEL2 is expressed at higher level in some primary human leukemia samples than in normal bone marrow. Furthermore, overexpression of TEL2 in NIH3T3-UCLA cells blocked the inhibitory effect of TEL1 on Ras-induced cellular transformation. These results suggest that TEL2 may play an important role in hematopoiesis and oncogenesis. PMID- 15342393 TI - Molecular imaging of temporal dynamics and spatial heterogeneity of hypoxia inducible factor-1 signal transduction activity in tumors in living mice. AB - Tumor hypoxia is a spatially and temporally heterogeneous phenomenon, which results from several tumor and host tissue-specific processes. To study the dynamics and spatial heterogeneity of hypoxia-inducible factor-1 (HIF-1)-specific transcriptional activity in tumors, we used repetitive noninvasive positron emission tomography (PET) imaging of hypoxia-induced HIF-1 transcriptional activity in tumors in living mice. This approach uses a novel retroviral vector bearing a HIF-1-inducible "sensor" reporter gene (HSV1-tk/GFP fusion) and a constitutively expressed "beacon" reporter gene (DsRed2/XPRT). C6 glioma cells transduced with this multireporter system revealed dose-dependent patterns in temporal dynamics of HIF-1 transcriptional activity induced by either CoCl2 or decreased atmospheric oxygen concentration. Multicellular spheroids of C6 reporter cells developed a hypoxic core when >350 microm in diameter. 18F-2' fluoro-2'deoxy-1beta-D-arabionofuranosyl-5-ethyl-uracil (FEAU) PET revealed spatial heterogeneity of HIF-1 transcriptional activity in reporter xenografts in mice as a function of size or ischemia-reperfusion injury. With increasing tumor diameter (>3 mm), a marked increase in HIF-1 transcriptional activity was observed in the core regions of tumors. Even a moderate ischemia-reperfusion injury in small C6 tumors caused a rapid induction of HIF-1 transcriptional activity, which persisted for a long time because of the inability of C6 tumors to rapidly compensate acute changes in tumor microcirculation. PMID- 15342394 TI - Hepatocyte growth factor/c-Met signaling promotes the progression of experimental human neuroblastomas. AB - Neuroblastoma is the most frequent solid childhood malignancy. Despite aggressive therapy, mortality is high due to rapid tumor progression to advanced stages. The molecules and mechanisms underlying poor prognosis are not well understood. Here, we report that cultured human neuroblastoma cells express the hepatocyte growth factor (HGF) and its receptor c-Met. Binding of HGF to c-Met triggers receptor autophosphorylation, indicating functional relevance of this interaction. HGF activates several downstream effectors of c-Met such as the mitogen-activated protein kinases extracellular signal-regulated kinase 1/extracellular signal regulated kinase 2 and phospholipase C-gamma, whereas signal transducer and activator of transcription 3 is constitutively activated in neuroblastoma cells expressing c-Met. In addition, HGF is able to stimulate expression and proteolytic activity of matrix metalloproteinase-2 and tissue-type plasminogen activator in neuroblastoma cells, thereby promoting degradation of extracellular matrix components. We show that HGF stimulates invasion of neuroblastoma cells in vitro and in vivo, and it promotes the formation of angiogenic neuroblastomas in vivo. These processes can be blocked by specific inhibitors of the mitogen activated protein kinase cascade, by inhibitors of phospholipase C-gamma, and also by the expression of a dominant negative signal transducer and activator of transcription 3 mutant. Our data provide the first evidence that the HGF/c-Met pathway is essential for invasiveness and malignant progression of human neuroblastomas. They further suggest that specific inhibitors of this pathway may be suitable as therapeutic agents to improve clinical outcome of neuroblastomas. PMID- 15342395 TI - Angiopoietin-3 inhibits pulmonary metastasis by inhibiting tumor angiogenesis. AB - Angiopoietins (Ang-1, Ang-2, and Ang-3) are the ligands of Tie-2 receptor tyrosine kinase. The essential roles of Ang-1 and Tie-2 in embryonic angiogenesis have been established, and studies have demonstrated the involvement of Ang-1 and Ang-2 in tumor angiogenesis. However, the role of Ang-3 in tumor angiogenesis and metastasis and the mechanism underlying its function are totally unknown. We have shown recently that Ang-3 is tethered on cell surface via heparan sulfate proteoglycans. In our current study, we have demonstrated that overexpression of Ang-3 inhibits pulmonary metastasis of Lewis lung carcinoma and TA3 mammary carcinoma (TA3) cells by inhibiting tumor angiogenesis and promoting apoptosis of the tumor cells. In addition, we have demonstrated that the binding of Ang-3 to the cell surface is required for the effective inhibition of Ang-3 on tumor metastasis and that Ang-3 inhibits endothelial cell proliferation and survival and blocks Ang-1- and vascular endothelial growth factor-induced activation of extracellular signal-regulated kinase 1/2 and Akt kinases, which likely underlie the Ang-3-mediated inhibition on tumor angiogenesis and metastasis. PMID- 15342396 TI - The ras/mitogen-activated protein kinase pathway inhibitor and likely tumor suppressor proteins, sprouty 1 and sprouty 2 are deregulated in breast cancer. AB - Sprouty (Spry) proteins were found to be endogenous inhibitors of the Ras/mitogen activated protein kinase pathway that play an important role in the remodeling of branching tissues. We investigated Spry expression levels in various cancers and found that Spry1 and Spry2 were down-regulated consistently in breast cancers. Such prevalent patterns of down-regulation may herald the later application of these isoforms as tumor markers that are breast cancer specific and more profound than currently characterized markers. Spry1 and 2 were expressed specifically in the luminal epithelial cells of breast ducts, with higher expression during stages of tissue remodeling when the epithelial ducts are forming and branching. These findings suggest that Sprys might be involved as a modeling counterbalance and surveillance against inappropriate epithelial expansion. The abrogation of endogenous Spry activity in MCF-7 cells by the overexpression of a previously characterized dominant-negative mutant of Spry, hSpry2Y55F resulted in enhanced cell proliferation in vitro. The hSpry2Y55F stably expressing cells also formed larger and greater number of colonies in the soft-agar assay. An in vivo nude mice assay showed a dramatic increase in the tumorigenic potential of hSpry2Y55F stable cells. The consistent down-regulation of Spry1 and 2 in breast cancer and the experimental evidence using a dominant-negative hSpry2Y55F indicate that Spry proteins may actively maintain tissue integrity that runs amok when their expression is decreased below normal threshold levels. This alludes to a previously unrecognized role for Sprys in cancer development. PMID- 15342397 TI - Utilization of bone marrow-derived endothelial cell precursors in spontaneous prostate tumors varies with tumor grade. AB - Id1 and Id3 genes are required for vascularization, growth, and metastasis of xenograft tumors. In Id-deficient mice, tumor transplantation and proangiogenic factors fail to mobilize and recruit circulating endothelial precursor cells (CEPs) and hematopoietic cells, leading to defective tumor angiogenesis in various models. To investigate the requirement of Id genes and bone marrow incorporation in spontaneous prostate tumors, we crossbred Id mutant mice with the transgenic adenocarcinoma of the mouse prostate (TRAMP) mice. Id1-/- Id3+/- TRAMP mice display delayed tumor growth at 24 weeks compared with wild-type TRAMP mice. Id1 and Id3 were strongly expressed in the endothelial cells of poorly differentiated prostate adenocarcinoma but not in the vasculature of well differentiated tumors, a finding that is corroborated in human prostate tumor samples. In Id-deficient TRAMP mice, the poorly differentiated tumors show extensive hemorrhage, whereas well-differentiated tumors exhibit none. Transplantation with Id wild-type bone marrow significantly reduced the hemorrhage in poorly differentiated prostate adenocarcinomas with bone marrow derived endothelial cells contributing to 14% of the tumor blood vessels. However, in well-differentiated prostate adenocarcinomas, there was little evidence of bone marrow-derived endothelial cell incorporation. These differences in the expression of Id genes, the effects of Id loss, and the recruitment of bone marrow-derived endothelial precursor cells in tumor vasculature between well differentiated and poorly differentiated prostate adenocarcinoma suggest that tumor angiogenesis varies depending on the tumor grade. PMID- 15342398 TI - Progressive loss of malignant behavior in telomerase-negative tumorigenic adrenocortical cells and restoration of tumorigenicity by human telomerase reverse transcriptase. AB - Replicative senescence/crisis is thought to act as a tumor suppressor mechanism. Although recent data indicate that normal human cells cannot be converted into cancer cells without telomerase, the original concept of senescence as a tumor suppressor mechanism is that senescence/crisis would act to limit the growth of telomerase-negative tumors. We show here that this concept is valid when oncogene expressing human and bovine cells are introduced into immunodeficient mice using tissue reconstruction techniques, as opposed to conventional subcutaneous injection. Primary human and bovine adrenocortical cells were transduced with retroviruses encoding Ha-Ras(G12V) and SV40 large T antigen and transplanted in immunodeficient mice using tissue reconstruction techniques. Transduced cells were fully malignant (invasive and metastatic) in this model. They had negligible telomerase activity both before transplantation and when recovered from tumors. When serially transplanted, tumors showed progressively slower growth, decreased invasion and metastasis, shortened telomeres, and morphological features of crisis. Whereas telomerase was not essential for malignant behavior, expression of human telomerase reverse transcriptase enabled cells from serially transplanted tumors that had ceased growth to reacquire tumorigenicity. Moreover, telomerase-negative oncogene-expressing cells were tumorigenic only when transplanted using tissue reconstruction techniques; human telomerase reverse transcriptase was required for cells to form tumors when cells were injected subcutaneously. This work provides a new model to study crisis in an in vivo setting and its effects on malignancy; despite having invasive and metastatic properties, cells are eventually driven into crisis by proliferation in the absence of a telomere maintenance mechanism. PMID- 15342399 TI - Proteolytic enzymes and altered glycosylation modulate dystroglycan function in carcinoma cells. AB - Alterations in the basement membrane receptor dystroglycan (DG) are evident in muscular dystrophies and carcinoma cells and characterized by a selective loss or modification of the extracellular alpha-DG subunit. Defects in posttranslational modifications of DG have been identified in some muscular dystrophies, but the underlying modifications in carcinoma cells have not yet been defined. We reveal here multiple posttranslational modifications that modulate the composition and function of DG in normal epithelial cells and carcinoma cells. We show that alpha DG is shed from the cell surface of normal and tumorigenic epithelial cells through a proteolytic mechanism that does not require direct cleavage of either alpha- or beta-DG. Shedding is dependent on metalloprotease activity and the proprotein convertase furin. Surprisingly, furin is also found to directly process alpha-DG as a proprotein substrate, changing the existing model of DG composition. We also show that the glycosylation of alpha-DG is altered in invasive carcinoma cells, and this modification causes complete loss of laminin binding properties. Together, these data elucidate several novel events regulating the functional composition of DG and reveal defects that arise during cancer progression, providing direction for efforts to restore this link with the basement membrane in carcinoma cells. PMID- 15342400 TI - Role of carbonic anhydrase IX in human tumor cell growth, survival, and invasion. AB - Carbonic anhydrase IX (CAIX) is a membrane-associated carbonic anhydrase (CA), strongly induced by hypoxia. CAIX is overexpressed in a variety of tumor types and associated with increased metastasis and poor prognosis. An inhibitor of CAs, acetazolamide has been reported to inhibit invasion. We used RNA interference (RNAi) to examine the function of CAIX in MDA468 and MDA231 breast carcinoma cells, which express high levels of CAIX under hypoxia. Hypoxia-induced CA activity was completely blocked by specific RNAi (P < 0.01). RNAi-treated cells showed growth delay in dense monolayer culture and a 50% reduction in clonogenic survival under hypoxia. In the MDA468 cells, there was no effect of RNAi treatment on invasion. In a cell line that did not induce CAIX under hypoxia, RT112, we found no effect on the ability of cells transfected with CAIX to invade or migrate. Thus, CAIX plays an important role in the growth and survival of tumor cells under normoxia and hypoxia, making it a potential target for cancer therapy, but is not involved in invasion. PMID- 15342401 TI - SRC family kinases mediate epidermal growth factor receptor ligand cleavage, proliferation, and invasion of head and neck cancer cells. AB - Head and neck squamous cell carcinomas (HNSCCs) are characterized by up regulation of the epidermal growth factor receptor (EGFR). We previously reported that a gastrin-releasing peptide/gastrin-releasing peptide receptor (GRP/GRPR) autocrine growth pathway is activated early in HNSCC carcinogenesis. GRP can induce rapid phosphorylation of EGFR and p42/44 mitogen-activated protein kinase (MAPK) activation in part via extracellular release of transforming growth factor alpha (TGF-alpha) by matrix metalloproteinases (MMPs). It has been reported that Src family kinases are activated by G-protein-coupled receptors (GPCRs), followed by downstream EGFR and MAPK activation. To further elucidate the mechanism of activation of EGFR by GRP in HNSCC, we investigated the role of Src family kinases. Blockade of Src family kinases using an Src-specific tyrosine kinase inhibitor A-419259 decreased GRP-induced EGFR phosphorylation and MAPK activation. GRP also failed to induce MAPK activation in dominant-negative c-Src transfected HNSCC cells. Invasion and growth assays showed that c-Src was required for GRP-induced proliferation or invasion of HNSCC cells. In addition to TGF-alpha release, GRP induced amphiregulin, but not EGF, secretion into HNSCC cell culture medium, an effect that was blocked by the MMP inhibitor marimastat. TGF-alpha and amphiregulin secretion by GRP stimulation also was inhibited by blockade of Src family kinases. These results suggest that Src family kinases contribute to GRP-mediated EGFR growth and invasion pathways by facilitating cleavage and release of TGF-alpha and amphiregulin in HNSCC. PMID- 15342402 TI - Intracellular clusterin induces G2-M phase arrest and cell death in PC-3 prostate cancer cells1. AB - Enhanced clusterin gene expression has been related frequently to organ remodeling, tissue involution, and cell death. Whether clusterin represents a leading cause or a consequence of apoptosis induction is still a matter of debate. Clusterin is known as an extracellular secreted glycoprotein in the mature form. However, truncated isoforms of the protein and nuclear localization of clusterin have been described recently in association to cell death. Here, we show the biological effects triggered in PC-3 androgen-independent prostate cancer cells by overexpression of an intracellular, not secreted form of clusterin (intracellular-clusterin). Transient transfection of PC-3 cells with intracellular-clusterin resulted in nuclear localization signal-independent massive nuclear localization of the protein leading to G2-M phase blockade followed by caspase-dependent apoptosis. Constitutive expression of intracellular clusterin (pFLAG- intracellular-clusterin) in recombinant PC-3 cells caused clonogenic toxicity. The rare pFLAG-intracellular clusterin surviving clones showed inhibition of the proliferation rate and altered phenotype with impaired mitosis and endoreduplication. In these cells, caspase-independent cell death was induced. Impaired cell cycle progression in pFLAG-intracellular-clusterin clones was associated to arrest at the G2-M checkpoint by down-regulation of the mitotic complex cyclin B1/cyclin-dependent kinase 1. Intriguingly, intracellular clusterin was localized exclusively in the cytoplasm in stably transfected cells, suggesting a negative correlation between nuclear clusterin accumulation and cell survival. These findings may possibly explain the conflicting results obtained in different laboratories, suggesting that clusterin might be a proapoptotic or a survival gene, also opening new perspectives for the characterization of androgen independent and apoptosis-resistant prostate cancer cells. PMID- 15342403 TI - Quantifying transient hypoxia in human tumor xenografts by flow cytometry. AB - Transient hypoxia is a poorly understood and potentially important factor that may limit tumor response to various forms of therapy. We assessed transient hypoxia on a global scale in two different human tumor xenografts by sequentially administering two hypoxia markers followed by quantification of hypoxic cells using flow cytometry. High levels of the first hypoxia marker (pimonidazole) were maintained in the circulation over an 8-hour period by multiple hourly injections, providing a "time-integrated" hypoxia measure showing an asymptotic increase in the total number of hypoxic cells. Subsequent administration of a second hypoxia marker (CCI-103F) showed that substantial numbers of the previously pimonidazole-labeled cells were no longer hypoxic during the circulation lifetime of the second marker. The overall fraction of tumor cells that demonstrated changes in hypoxic status with time increased with different kinetics and by different magnitudes in the two xenograft systems. Specifically, up to 20% of the cells in SiHa (human cervical squamous cell carcinoma) tumors and up to 8% of the cells in WiDr (human colon adenocarcinoma) tumors were intermittently hypoxic over an 8-hour period. Also, the tumor cells that demonstrated transient hypoxia were typically not adjacent to functional tumor blood vessels. Similar approaches could be used in the clinic to provide information on the duration of intermittent hypoxia episodes and the fraction of transiently hypoxic tumor cells, which would, in turn, have important implications for the strategic improvement of cancer therapy. PMID- 15342404 TI - Enhanced redundancy in Akt and mitogen-activated protein kinase-induced survival of malignant versus normal prostate epithelial cells. AB - Activation of the downstream akt and mitogen-activated protein kinases is associated with development and progression of prostate cancer to the lethal androgen-independent state. However, the causal role of these downstream kinases in androgen-independent prostate cancers is unknown. In this study, activation and requirements of akt and mitogen-activated protein kinase (erk, p38, and jnk) signaling for the survival and proliferation of five malignant human cell lines encompassing the spectrum of androgen-independent prostate cancers was compared with the activation and requirements in normal prostate epithelial cells. Using Western blotting with phospho-antibodies, we detected differential activation in exponentially growing, growth factor-deprived, and restimulated cultures of malignant versus normal cells. The inhibition of erk, p38, jnk, and akt with U0126, SB203580, SP600125, and Akt inhibitor, respectively, document that normal cells require simultaneous erk and jnk signaling for survival, plus akt signaling for proliferation. In malignant cells, however, only jnk inhibition as monotherapy produces a consistent apoptotic response, although the combinatorial inhibition of jnk, erk, p38 plus akt results in statistically enhanced apoptosis. These results demonstrate that prostate cancer progression to a lethal androgen independent state involves the acquisition of an enhanced redundancy in downstream survival signaling. PMID- 15342405 TI - A single treatment of yttrium-90-labeled CHX-A"-C6.5 diabody inhibits the growth of established human tumor xenografts in immunodeficient mice. AB - Antitumor diabody molecules are noncovalent single-chain Fv dimers that recapitulate the divalent binding properties of native IgG antibodies. Diabodies are capable of substantial accumulation in tumor xenografts expressing relevant antigens in immunodeficient mouse models. With a Mr of approximately 55,000, diabodies are rapidly cleared from the circulation, resulting in tumor-to-blood ratios that significantly exceed those achieved early after the administration of monoclonal antibodies. We have evaluated the therapeutic potential of the beta emitting isotope yttrium-90 (t1/2, 64 hours) conjugated to the C6.5K-A diabody that specifically targets the HER2/neu human tumor-associated antigen. We have found that a single intravenous dose of 150 microCi (200 microg) 90Y-CHX-A"-C6.5K A diabody substantially inhibits the growth rates of established MDA-361/DYT2 human breast tumor xenografts in athymic nude mice. In contrast, 300 microCi (300 microg) 90Y-CHX-A"-C6.5K-A diabody resulted in only a minor delay in the growth of SK-OV-3 human ovarian cancer xenografts. The maximum tolerated dose was also dependent on the tumor xenograft model used. These studies indicate that genetically engineered antitumor diabody molecules can be used as effective vehicles for radioimmunotherapy. PMID- 15342406 TI - Ibuprofen inhibits survival of bladder cancer cells by induced expression of the p75NTR tumor suppressor protein. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are used to reduce inflammation and as analgesics by inhibition of cyclooxygenase-2. At higher concentrations, some NSAIDs inhibit proliferation and induce apoptosis of cancer cells. Although several molecular mechanisms have been postulated to explain the anticancer effects of NSAIDs, they do not involve merely the inhibition of cyclooxygenase-2, and a more proximate initiator molecule may be regulated by NSAIDs to inhibit growth. The p75 neurotrophin receptor (p75NTR) is a proximate cell membrane receptor glycoprotein that has been identified as a tumor and metastasis suppressor. We observed that NSAID treatment of cell lines from bladder and other organs induced expression of the p75NTR protein. Of the different types of NSAIDs examined, ibuprofen was more efficacious than aspirin and acetaminophen and comparable with (R)-flurbiprofen and indomethacin in induction of p75NTR protein expression. This rank order NSAID induction of the p75NTR protein correlated with the ability of these NSAIDs to reduce cancer cell survival. To examine a mechanistic relationship between ibuprofen induction of p75NTR protein and inhibition of survival, bladder cancer cells were transfected with ponasterone A inducible vectors that expressed a death domain-deleted (DeltaDD) or intracellular domain-deleted (DeltaICD) p75NTR product that acts as a dominant negative antagonist of the intact p75NTR protein. Expression of DeltaDD and DeltaICD rescued cells from ibuprofen inhibition of growth. These observations suggest that p75NTR is an important upstream modulator of the anticancer effects of NSAIDs and that ibuprofen induction of the p75NTR protein establishes an alternate mechanism by which ibuprofen may exert an anticancer effect. PMID- 15342407 TI - Enhancement of the efficacy of chemotherapy for lung cancer by simultaneous suppression of multidrug resistance and antiapoptotic cellular defense: novel multicomponent delivery system. AB - The efficacy of chemotherapy of lung cancer is limited by the development of resistance in cancer cells during treatment. In most lung cancers, this resistance is associated with the overexpression of (a) multidrug resistance associated protein (MRP) responsible for drug efflux from the cancer cells (pump resistance) and (b) BCL2 protein that activates antiapoptotic cellular defense (nonpump resistance). A novel liposomal proapoptotic anticancer drug delivery system was developed to enhance anticancer efficacy of the well-established drug doxorubicin (DOX). This multicomponent drug delivery system was tested on multidrug-sensitive and -resistant human small-cell lung cancer cells. The drug delivery system includes four components: (a) liposome as a carrier, (b) DOX as an inductor of apoptosis, (c) antisense oligonucleotides (ASOs) targeted to MRP1 mRNA as a suppressor of pump resistance, and (d) ASOs targeted to BCL2 mRNA as a suppressor of nonpump resistance. Intracellular internalization of ASOs and DOX; the influence of the proposed system on the expression of genes and proteins involved in the multidrug resistance, cytotoxicity, and apoptosis induction and antiapoptotic defense; and the activity of caspases were studied. It was found that the proposed liposomal delivery system successfully delivered ASOs and DOX to cell nuclei, inhibited MRP1 and BCL2 protein synthesis, and substantially increased the anticancer action of DOX by stimulating the caspase-dependent pathway of apoptosis in multidrug-resistant human lung cancer cells. PMID- 15342408 TI - Hyperforin inhibits cancer invasion and metastasis. AB - Hyperforin (Hyp), the major lipophilic constituent of St. John's wort, was assayed as a stable dicyclohexylammonium salt (Hyp-DCHA) for cytotoxicity and inhibition of matrix proteinases, tumor invasion, and metastasis. Hyp-DCHA triggered apoptosis-associated cytotoxic effect in both murine (C-26, B16-LU8, and TRAMP-C1) and human (HT-1080 and SK-N-BE) tumor cells; its effect varied, with B16-LU8, HT-1080, and C-26 the most sensitive (IC50 = 5 to 8 micromol/L). At these concentrations, a marked and progressive decline of growth was observed in HT-1080 cells, whereas untransformed endothelial cells were only marginally affected. Hyp-DCHA inhibited in a dose-dependent and noncompetitive manner various proteinases instrumental to extracellular matrix degradation; the activity of leukocyte elastase was inhibited the most (IC50 = 3 micromol/L), followed by cathepsin G and urokinase-type plasminogen activator, whereas that of the matrix metalloproteinases (MMPs) 2 and 9 showed an IC50 > 100 micromol/L. Nevertheless, inhibition of extracellular signal-regulated kinase 1/2 constitutive activity and reduction of MMP-2 and MMP-9 secretion was triggered by 0.5 micromol/L Hyp-DCHA to various degrees in different cell lines, the most in C 26. Inhibition of C-26 and HT-1080 cell chemoinvasion (80 and 54%, respectively) through reconstituted basement membrane was observed at these doses. Finally, in mice that received i.v. injections of C-26 or B16-LU8 cells, daily i.p. administration of Hyp-DCHA-without reaching tumor-cytotoxic blood levels remarkably reduced inflammatory infiltration, neovascularization, lung weight ( 48%), and size of experimental metastases with C-26 (-38%) and number of lung metastases with B16-LU8 (-22%), with preservation of apparently healthy and active behavior. These observations qualify Hyp-DCHA as an interesting lead compound to prevent and contrast cancer spread and metastatic growth. PMID- 15342409 TI - Role of p53 in sensing oxidative DNA damage in response to reactive oxygen species-generating agents. AB - The tumor suppressor p53 plays an important role in the regulation of cellular response to DNA damage. Recent studies suggest that p53 is able to bind DNA with certain structural alterations in a sequence-independent manner and to interact with several molecules involved in DNA repair. This study was undertaken to test the hypothesis that p53 may participate in sensing oxidative DNA damage, the most frequently occurring spontaneous DNA lesion, and modulate its repair by the base excision repair (BER) machinery. Using synthetic DNA containing 8-oxo-7,8-dihydro 2'-deoxyguanosine (8-oxoG), we showed that p53 was pulled down together with two BER proteins, human 8-oxoguanine glycosylase (hOGG1) and AP endonuclease (APE). Functional analysis showed that p53 significantly enhanced the sequential activities of hOGG1 and APE in excising the 8-oxoG nucleotide from DNA in vitro. The ability of p53 to enhance the removal of oxidized DNA bases was further demonstrated in vivo using a pair of p53 isogenic lines. HCT116 p53+/+ cells exhibit a more rapid removal of 8-oxoG from DNA than p53-/- cells exposed to the same levels of reactive oxygen species (ROS) stress. Together, these results suggest that p53 participates in sensing oxidative DNA damage and modulates BER function in response to persistent ROS stress. PMID- 15342410 TI - Activation of nuclear factor kappaB In vivo selectively protects the murine small intestine against ionizing radiation-induced damage. AB - Exposure of mice to total body irradiation induces nuclear factor kappaB (NFkappaB) activation in a tissue-specific manner. In addition to the spleen, lymph nodes, and bone marrow, the tissues that exhibit NFkappaB activation now include the newly identified site of the intestinal epithelial cells. NFkappaB activated by total body irradiation mainly consists of NFkappaB p50/RelA heterodimers, and genetically targeted disruption of the NFkappaB p50 gene in mice significantly decreased the activation. By comparing tissue damage and lethality in wild-type and NFkappaB p50 knockout (p50-/-) mice after they were exposed to increasing doses of total body irradiation, we additionally examined the role of NFkappaB activation in total body irradiation-induced tissue damage. The results show that p50-/- mice are more sensitive to total body irradiation induced lethality than wild-type mice (LD50/Day 7: wild-type = 13.12 Gy versus p50-/- = 7.75 Gy and LD50/Day 30: wild-type = 9.31 Gy versus p50-/- = 7.81 Gy). The increased radiosensitivity of p50-/- mice was associated with an elevated level of apoptosis in intestinal epithelial cells and decreased survival of the small intestinal crypts compared with wild-type mice (P < 0.01). In addition, RelA/TNFR1-deficient (RelA/TNFR1-/-) mice also exhibited a significant increase in intestinal epithelial cell apoptosis after they were exposed to total body irradiation as compared with TNFR1-deficient (TNFR1-/-) mice (P < 0.01). In contrast, no significant increase in total body irradiation-induced apoptosis or tissue injury was observed in bone marrow cells, spleen lymphocytes, and the liver, heart, lung, and kidney of p50-/- mice in comparison with wild-type mice. These findings indicate that activation of NFkappaB selectively protects the small intestine against ionizing radiation-induced damage. PMID- 15342411 TI - Detection of invasive colon cancer using a novel, targeted, library-derived fluorescent peptide. AB - Sensitive methods to detect the earliest forms of colorectal cancers remain a challenge despite the development of serum and stool biomarkers. We reasoned that fluorescent affinity ligands derived from library screens can be developed to improve the detection and localization of early malignant lesions by endoscopy. We have developed an imaging agent for real-time endoscopic tumor detection in a murine model using a previously identified phage library-derived colon cancer specific cyclic peptide and fluorescent moieties. The modified peptide had a 24 minute blood half life and tumoral accumulation was 6.9% of injected dose/g, approximately 7-fold higher than a scrambled control peptide. Orthotopic colonic tumors (HT29) were readily detectable by fluorescence endoscopy even when tumors were submucosal. These results show proof-of-principle that disease-specific library-derived fluorescent probes can be rapidly developed for use in the early detection of cancers by optical means. PMID- 15342412 TI - Growth inhibition of human prostate cancer cells in human adult bone implanted into nonobese diabetic/severe combined immunodeficient mice by a ligand-specific antibody to human insulin-like growth factors. AB - Advanced prostate cancer frequently involves the bone that has the largest content of insulin-like growth factors (IGFs). However, the role of bone-derived IGFs in bone metastasis of prostate cancer has not been studied extensively because of the lack of a reliable animal model. Therefore, we investigated whether a novel antibody directed against human IGF-I and IGF-II (KM1468) could inhibit the development of new bone tumors and the progression of established bone tumors in nonobese diabetic/severe combined immunodeficient mice implanted with human adult bone. We first confirmed that KM1468 bound specifically to human IGF-I, human IGF-II, and mouse IGF-II but not to insulin. It also blocked autophosphorylation of the type I IGF receptor induced by the binding of IGFs in human-type I IGF receptor-overexpressing BALB/c 3T3 cells, and it inhibited the IGF-stimulated growth of MDA PCa 2b cells in vitro. Then mice were injected intraperitoneally with KM1468 once weekly for 4 weeks either immediately or 4 weeks after inoculation of MDA PCa 2b cells. KM1468 markedly and dose-dependently suppressed the development of new bone tumors and the progression of established tumor foci, as determined by histomorphometry, and it also decreased serum prostate-specific antigen levels, compared with the control. This is the first report of an IGF ligand-specific inhibitory antibody that suppresses the growth of human prostate cancer cells in human adult bone. These results indicate that the IGF signaling axis is a potential target for prevention and treatment of bone metastases arising from prostate cancer. PMID- 15342413 TI - Visualization of intrathoracically disseminated solid tumors in mice with optical imaging by telomerase-specific amplification of a transferred green fluorescent protein gene. AB - Currently available methods for detection of tumors in vivo such as X-ray, computed tomography, and ultrasonography are noninvasive and have been well studied; the images, however, are not specific for tumors. Direct optical imaging of tumor cells in vivo that can clearly distinguish them from surrounding normal tissues may be clinically useful. Here, we describe a new approach to visualizing tumors whose fluorescence can be detected using tumor-specific replication competent adenovirus (OBP-301, Telomelysin) in combination with Ad-GFP, a replication-deficient adenovirus expressing green fluorescent protein (GFP). Human telomerase reverse transcriptase is the catalytic subunit of telomerase, which is highly active in cancer cells but quiescent in most normal somatic cells. We constructed an adenovirus 5 vector in which the human telomerase reverse transcriptase promoter element drives expression of E1A and E1B genes linked with an internal ribosome entry site and showed that OBP-301 replicated efficiently in human cancer cells, but not in normal cells such as human fibroblasts. When the human lung and colon cancer cell lines were infected with Ad-GFP at a low multiplicity of infection, GFP expression could not be detected under a fluorescence microscope; in the presence of OBP-301, however, Ad-GFP replicated in these tumor cells and showed strong green signals. In contrast, coinfection with OBP-301 and Ad-GFP did not show any signals in normal cells such as fibroblasts and vascular endothelial cells. We also found that established subcutaneous tumors could be visualized after intratumoral injection of OBP-301 and Ad-GFP. A549 human lung tumors and SW620 human colon tumors transplanted into BALB/c nu/nu mice were intratumorally injected with 8 x 10(5) plaque-forming units of Ad-GFP in combination with 8 x 10(6) plaque-forming units of OBP-301. Within 3 days of treatment, the fluorescence of the expressed GFP became visible by a three-chip color cooled charged-coupled device camera in these tumors, whereas intratumoral injection of Ad-GFP alone could not induce GFP fluorescence. Moreover, intrathoracic administration of Ad-GFP and OBP-301 could visualize disseminated A549 tumor nodules in mice after intrathoracic implantation. Our results indicate that intratumoral or intrathoracic injection of Ad-GFP in combination with OBP-301 might be a useful diagnostic method that provides a foundation for future clinical application. PMID- 15342414 TI - Fas signaling is involved in the control of hair follicle response to chemotherapy. AB - Chemotherapeutic agents induce p53-dependent apoptosis in the hair follicle (HF) resulting in hair loss, a common side effect of cancer therapy. Here, we show that Fas as a p53 target plays important role in the HF response to cyclophosphamide. Specifically, we demonstrate that Fas is up-regulated in HF keratinocytes after cyclophosphamide treatment, Fas ligand-neutralizing antibody partially inhibits HF response to cyclophosphamide in wild-type mice, and Fas knockout mice show significant retardation of cyclophosphamide-induced HF involution associated with reduced Fas-associated death domain and caspase-8 expression. These data raise a possibility to explore blockade of Fas signaling as a part of complex local therapy for inhibiting keratinocyte apoptosis and hair loss induced by chemotherapy. PMID- 15342415 TI - Ceramide reduction and transcriptional up-regulation of glucosylceramide synthase through doxorubicin-activated Sp1 in drug-resistant HL-60/ADR cells. AB - Treatment with doxorubicin (DOX) induced apoptosis with an increase of ceramide content in drug-sensitive HL-60 cells, but not in drug-resistant HL-60/ADR cells. In HL-60/ADR cells (but not in HL-60 cells), the levels of mRNA, protein, and activity in glucosylceramide synthase (GCS), which converts ceramide to glucosylceramide, were up-regulated in response to DOX. Thus, abrogation of apoptosis in HL-60/ADR cells might be involved in ceramide reduction through DOX induced up-regulation of GCS function. Because we reported that a GC-rich/Sp1 promoter binding region was of importance in the regulation of GCS expression, the role of Sp1 in DOX-induced up-regulation of GCS and apoptosis was investigated. DOX induced Sp1 activation in HL-60/ADR cells, as assessed by Sp1 gel shift and promoter-luciferase reporter assays, whereas transfection of double stranded oligodeoxynucleotides (ODNs) containing a GC-rich/Sp1 region (Sp1 decoy ODNs) inhibited DOX-induced Sp1 activation. In addition, DOX-increased mRNA and enzyme activity in GCS were inhibited by Sp1 decoy, in conjunction with corresponding elevations of ceramide content. Moreover, DOX-induced apoptotic cell death was significantly increased in Sp1 decoy ODN-transfected HL-60/ADR cells over mock-transfected HL-60/ADR cells. Together, the results suggest that transcriptional up-regulation of GCS through DOX-induced activation of Sp1 is one potential mechanism to regulate ceramide increase and apoptosis in HL-60/ADR cells. PMID- 15342416 TI - Evaluation of human thymidine kinase 1 substrates as new candidates for boron neutron capture therapy. AB - Thymidine analogs containing o-carboranylalkyl groups at the 3-position were screened as potential substrates for human thymidine kinase 1 (TK1), an enzyme that is selectively expressed in a variety of rapidly proliferating cells, including tumor cells. On the basis of previous studies, 12 of these were identified as potential delivery agents for boron neutron capture therapy, a therapeutic method used for the treatment of high-grade brain tumors. Compound 4 with a pentylene spacer between the o-carborane cage and the thymidine scaffold and compound 10, which has an additional dihydroxypropyl substituent at the o carborane cage, were the best substrates for TK1 with kcat/Km values of 27% and 36% relative to that of thymidine, respectively. These compounds showed partial competitive inhibition for thymidine phosphorylation by TK1. Neither compound was a substrate of recombinant human thymidine phosphorylase nor were their respective 5'-monophosphates substrates of 5'-deoxynucleotidase 1, thereby indicating potential in vivo stability. The octanol/water partition coefficient for compound 10 was 2.09, suggesting that it has excellent physiochemical properties for crossing the blood brain barrier and penetrating brain tissue. The in vitro cytotoxic effect of the 12 analogs was moderate to low in mammalian cell cultures with IC50 values between 10 and 160 micromol/L. Compounds 4 and 10 were taken up selectively and retained by the murine fibroblast L929 cell line, in contrast to its TK1-deficient variant. These findings suggest that compound 10 is a promising candidate for selective delivery of boron-10 to malignant cells, and additional in vivo studies are planned to evaluate it for boron neutron capture therapy of brain tumors. PMID- 15342417 TI - Boron-containing nucleosides as potential delivery agents for neutron capture therapy of brain tumors. AB - The purpose of the present study was to evaluate both in vitro and in vivo a series of boron-containing nucleosides that potentially could be used as delivery agents for neutron capture therapy. The rationale for their synthesis was based on the fact that proliferating neoplastic cells have increased requirements for nucleic acid precursors, and, therefore, they should preferentially localize in the tumor. A series of 3-carboranlyalkyl thymidine analogs has been synthesized and a subset, designated N4, N5, and N7, and the corresponding 3-dihydroxypropyl derivatives, designated N4-2OH, N5-2OH, and N7-2OH, have been selected for evaluation. Using these compounds as substrates for recombinant human thymidine kinase-1 and the mitochondrial isoenzyme thymidine kinase-2, the highest phosphorylation levels relative to thymidine were seen with N5 and the corresponding dihydroxypropyl analog N5-2OH. In contrast, N4, N4-OH, N7, and N7 OH had substantially lower phosphorylation levels. To compare compounds with high and low thymidine kinase-1 substrate activity, N5 and N7 and the corresponding dihydroxypropyl derivatives were selected for evaluation of their cellular toxicity, uptake and retention by the F98 rat glioma, human MRA melanoma, and murine L929 cell lines, all of which are thymidine kinase-1(+), and a mutant L929 cell line that is thymidine kinase-1(-). N5-2OH was the least toxic (IC50, 43-70 microm), and N7 and N7-2OH were the most toxic (IC50, 18-49 microm). The highest boron uptake was seen with N7-2OH by the MRA 27 melanoma and L929 wild-type (wt) cell lines. The highest retention was seen with L929 (wt) cells, and this ranged from 29% for N5-2OH to 46% for N7. Based on the in vitro toxicity and uptake data, N5-2OH was selected for in vivo biodistribution studies either in rats bearing intracerebral implants of the F98 glioma or in mice bearing either s.c. or intracerebral implants of L929 (wt) tumors. At 2.5 hours after convection enhanced delivery, the boron values for the F98 glioma and normal brain were 16.2 +/- 2.3 and 2.2 microg/g, respectively, and the tumor to brain ratio was 8.5. Boron values at 4 hours after convection-enhanced delivery of N5-2OH to mice bearing intracerebral implants of L929 (wt) or L929 thymidine kinase-1(-) tumors were 39.8 +/- 10.8 and 12.4 +/- 1.6 microg/g, respectively, and the corresponding normal brain values were 4.4 and 1.6 microg/g, thereby indicating that there was selective retention by the thymidine kinase-1(+) tumors. Based on these favorable in vitro and in vivo data, neutron capture therapy studies will be initiated using N5-2OH in combination with two non-cell cycle dependent boron delivery agents, boronophenylalanine and sodium borocaptate. PMID- 15342418 TI - P21Cip1 is a critical mediator of the cytotoxic action of thymidylate synthase inhibitors in colorectal carcinoma cells. AB - We have demonstrated previously that interferon (IFN)-gamma sensitizes human colon carcinoma cell lines to the cytotoxic effects of 5-fluorouracil combined with leucovorin and to the thymidylate synthase inhibitor, ZD9331, dependent on thymineless stress-induced DNA damage, independent of p53. Here we demonstrate that the cyclin-dependent kinase (CDK) inhibitor p21(Cip1) regulates thymineless stress-induced cytotoxicity in these cells. HCT116 wild-type (wt) and p53-/- cells underwent apoptosis and loss in clonogenic survival when exposed to ZD9331, whereas p21Cip1-/- cells were resistant. In contrast, IFN-gamma induced marked cytotoxicity in p21Cip1-/- cells only. ZD9331 induced p21Cip1 up-regulation in all of the cell lines examined, as did thymidine deprivation in thymidylate synthase-deficient (thymidylate synthase-) cells. Furthermore, selective induction of p21Cip1 in RKO was sufficient to induce apoptosis. P21Cip1, cdk1, cdk2, and cyclin E mRNA expression increased coincident with S-phase accumulation in HT29 cells treated with ZD9331 or 5fluorouracil/leucovorin, as demonstrated by cDNA microarray analyses. Cell cycle analyses revealed that HCT116 wt and p21Cip1 -/- cells accumulated in S phase within 24 h of ZD9331 exposure; however, wt cells exited S-phase more rapidly, where apoptosis occurred before mitosis, either in late S or G2. Finally, the CDK inhibitor roscovitine potentiated the cytotoxic activity of ZD9331 in both wt and p21Cip1-/- cells, strongly suggesting a role for p21Cip1-dependent CDK inhibition in cytotoxicity induced by thymidylate synthase inhibition. In summary, p21Cip1 positively regulates the cytotoxic action of thymidylate synthase inhibitors, negatively regulates the cytotoxic action of IFN-gamma, and enhances S-phase exit after thymineless stress, possibly via interaction with CDK-cyclin complexes. PMID- 15342419 TI - Direct assessment of drug penetration into tissue using a novel application of three-dimensional cell culture. AB - The failure of many anticancer drugs to control growth of solid cancers may stem in part from inadequate delivery to tumor regions distant from vasculature. Although the identification of new anticancer drug targets has led to the development of many new drug candidates, there is a lack of methodology for identifying drugs that adequately penetrate tumor tissue. We have developed a novel multilayered cell culture-based assay, which detects the penetration of anticancer drugs based on their effect within tissue. Drug exposures are made over 1 hour to one side of a disk of tissue approximately 150-microm thick, with the other side temporarily closed off, and penetration is then assessed 1-3 days later via bromodeoxyuridine-based detection of S-phase cells. Using this assay, the tissue distribution of a selection of anthracycline analogues was assessed. At clinically relevant exposures, none of the agents were able to affect cells on the far side of the culture at levels approaching that seen on the near (exposed) side. Doxorubicin and epirubicin exhibited approximately 10-fold decreases in the drug exposure seen by the cells on the far side relative to those on the near side of the cultures, whereas for daunorubicin and mitoxantrone, approximately 30 fold and >30-fold decreases were observed respectively. Results were consistent with the observed gradients in drug-derived fluorescence of doxorubicin, epirubicin, and daunorubicin. This model could be applied as a simple anticancer drug development screen to discover drugs that exhibit desirable penetration properties. PMID- 15342420 TI - Expression of complement factor H by lung cancer cells: effects on the activation of the alternative pathway of complement. AB - The complement system is important in immunosurveillance against tumors. However, malignant cells are usually resistant to complement-mediated lysis. In this study, we examine the expression of factor H, an inhibitor of complement activation, and factor H-like protein 1 (FHL-1), its alternatively spliced form, in lung cancer. We also evaluate the potential effect of factor H/FHL-1 in the protection of lung cancer cells against the activation of the complement cascade. By Northern blot analysis we demonstrate a high expression of factor H and FHL-1 in most non-small cell lung cancer cell lines, although neuroendocrine pulmonary tumors (small cell lung carcinoma and carcinoid cell lines) had undetectable levels. Western blot analysis of conditioned medium showed the active secretion of factor H and FHL-1 by cells that were positive by Northern blot. Expression of factor H/FHL-1 mRNA was also shown in a series of non-small cell lung cancer biopsies by in situ hybridization. Interestingly, many cultured lung cancer cells were able to bind fluorescence-labeled factor H to their surfaces. Deposition of C3 fragments from normal human serum on H1264, a lung adenocarcinoma cell line, was more efficient when factor H/FHL-1 activity was blocked by specific antibodies. Blocking factor H/FHL-1 activity also enhanced the release of anaphylatoxin C5a and moderately increased the susceptibility of these cells to complement-mediated cytotoxicity. In summary, we demonstrate the expression of factor H and FHL-1 by some lung cancer cells and analyze the contribution of these proteins to the protection against complement activation. PMID- 15342421 TI - High frequency of functionally active Melan-a-specific T cells in a patient with progressive immunoproteasome-deficient melanoma. AB - Tumor-reactive T cells play an important role in cancer immunosurveillance. Applying the multimer technology, we report here an unexpected high frequency of Melan-A-specific CTLs in a melanoma patient with progressive lymph node metastases, consisting of 18 and 12.8% of total peripheral blood and tumor infiltrating CD8+ T cells, respectively. Melan-A-specific CTLs revealed a high cytolytic activity against allogeneic Melan-A-expressing target cells but failed to kill the autologous tumor cells. Loading of the tumor cells with Melan-A peptide reversed the resistance to killing, suggesting impaired function of the MHC class I antigen processing and presentation pathway. Mutations of the coding region of the HLA-A2 binding Melan-A26-35 peptide or down-regulation of the MHC class I heavy chain, the antigenic peptide TAP, and tapasin could be excluded. However, PCR and immunohistochemical analysis revealed a deficiency of the immunoproteasomes low molecular weight protein 2 and low molecular weight protein 7 in the primary tumor cells, which affects the quantity and quality of generated T-cell epitopes and might explain the resistance to killing. This is supported by our data, demonstrating that the resistance to killing can be partially reversed by pre-exposure of the tumor cells to IFN-gamma, which is known to induce the immunoproteasomes. Overall, this is the first report of an extremely high frequency of tumor-specific CTLs that exhibit competent T-cell-effector functions but fail to lyse the autologous tumor cells. Immunotherapeutic approaches should not only focus on the induction of a robust antitumor immune response, but should also have to target tumor immune escape mechanisms. PMID- 15342422 TI - Bone marrow stroma influences transforming growth factor-beta production in breast cancer cells to regulate c-myc activation of the preprotachykinin-I gene in breast cancer cells. AB - Breast cancer cells (BCCs) have preference for the bone marrow (BM). This study used an in vitro coculture of BCCs and BM stroma to represent a model of early breast cancer metastasis to the BM. The overarching hypothesis states that once BCCs are in the BM, microenvironmental factors induce changes in the expression of genes for cytokines and preprotachykinin-I (PPT-I) in both BCCs and stromal cells. Consequently, the expression of both PPT-I and cytokines are altered to facilitate BCC integration within BM stroma. Cytokine and transcription factor arrays strongly suggested that transforming growth factor-beta (TGF-beta) and c myc regulate the expression of PPT-I so as to facilitate BCC integration among stroma. Northern analyses and TGF-beta bioassays showed that stromal cells and BCCs influence the level of PPT-I and TGF-beta in each other. In cocultures, PPT I and TGF-beta expressions were significantly (P < 0.05) increased and decreased, respectively. TGF-beta and PPT-I were undetectable in separate stromal cultures but were expressed as cocultures. Two consensus sequences for c-myc in the 5' flanking region of the PPT-I gene were shown to be functional using gel shift and reporter gene assays. Mutagenesis of c-myc sites, neutralization studies with anti-TGF-beta, and transient tranfections all showed that c-myc is required for TGF-beta-mediated induction of PPT-I in BCCs. TGF-beta was less efficient as a mediator of BCC integration within stroma for c-myc-BCCs. Because the model used in this study represents BCC integration within BM stroma, these studies suggest that TGF-beta is important to the regulation of PPT-I in the early events of bone invasion by BCCs. PMID- 15342423 TI - High-dose granulocyte-macrophage colony-stimulating factor-producing vaccines impair the immune response through the recruitment of myeloid suppressor cells. AB - Tumor vaccines have shown promise in early clinical trials. Among them, tumor cells genetically engineered to secrete biologically active granulocyte macrophage colony-stimulating factor (GM-CSF) can generate a systemic antitumor immune response. Although the minimal required GM-CSF dose produced by modified tumor cells to achieve a measurable antitumor effect is well known, no data examined whether an upper therapeutic limit may exist for this vaccination strategy. Because recent data demonstrate an immunosuppressive effect of GM-CSF produced by growing tumors, we thus sought to determine whether high GM-CSF doses administered in a vaccine formulation could impair antitumor immunity. Using a vaccine strategy involving a GM-CSF-producing bystander cell line (B78H1-GM) admixed with autologous tumor, we assessed the impact of varying doses of GM-CSF while maintaining a constant antigen dose. Our results defined a threshold above which a GM-CSF-based vaccine not only lost its efficacy, but more importantly for its clinical implications resulted in substantial immunosuppression in vivo. Above this threshold, GM-CSF induced Gr1+/CD11b+ myeloid suppressor cells that substantially impaired antigen-specific T-cell responses and adversely affected antitumor immune responses in vivo. The dual effects of GM-CSF are mediated by the systemic and not local concentration of this cytokine. Myeloid suppressor cell-induced immunosuppression is mediated by nitric oxide production via inducible nitric oxide synthase (iNOS) because the specific iNOS inhibitor, l NMMA, restored antigen-specific T-cell responsiveness in vitro. Taken together, our data demonstrated the negative impact of supra-therapeutic vaccine doses of GM-CSF and underscored the importance of identifying these critical variables in an effort to increase the therapeutic efficacy of tumor vaccines. PMID- 15342424 TI - The ADPRT V762A genetic variant contributes to prostate cancer susceptibility and deficient enzyme function. AB - The ADP-ribosyltransferase (ADPRT) gene encodes a zinc-finger DNA-binding protein, poly(ADP-ribose) polymerase-1 (PARP-1), that modifies various nuclear proteins by poly(ADP-ribosyl)ation and functions as a key enzyme in the base excision repair pathway. We have conducted two studies to test whether an amino acid substitution variant, ADPRT V762A (T2444C), is associated with prostate cancer (CaP) risk and decreased enzyme function. The first study used genomic DNA samples from an ongoing, clinic-based case-control study (488 cases and 524 controls) to show that a higher percentage of the CaP cases carried the ADPRT 762 AA genotype than controls (4% versus 2%). In Caucasians, the AA genotype was significantly associated with increased CaP risk [odds ratio (OR), 2.65; 95% confidence interval (CI), 1.08-6.49], and the VA genotype was associated with a slight but not significantly increased CaP risk (OR, 1.18; 95% CI, 0.85-1.64) using VV as the referent group after adjustment for age, benign prostatic hyperplasia, and family history. Furthermore, this association was stronger in younger (<65) men (OR, 4.77; 95% CI, 1.01-22.44) than older (> or =65) men (OR, 1.78; 95% CI, 0.55-5.82). The second study used freshly isolated peripheral lymphocytes from 354 cancer-free subjects to demonstrate that the ADPRT 762 A allele contributed to significantly lower adenosine diphosphate ribosyl transferase (ADPRT)/PARP-1 activities in response to H2O2 in a gene dosage dependent manner (P < 0.0001, test for linear trend). The PARP-1 activities (mean +/- SD dpm/10(6) cells) were 18,554 +/- 9,070 (n=257), 14,847 +/- 7,082 (n=86), and 12,155 +/- 6,334 (n=11) for VV, VA, and AA genotypes, respectively. This study is the first to provide evidence that the ADPRT V762A-genetic variant contributes to CaP susceptibility and altered ADPRT/PARP-1 enzyme function in response to oxidative damage. PMID- 15342425 TI - Silibinin protects against photocarcinogenesis via modulation of cell cycle regulators, mitogen-activated protein kinases, and Akt signaling. AB - Here, we assessed the protective effect of silibinin on UVB-induced skin carcinogenesis in SKH-1 hairless mice. Topical application of silibinin before or immediately after UVB exposure or its dietary feeding resulted in a strong protection against photocarcinogenesis, in terms of tumor multiplicity (60-66%; P < 0.001), tumor volume per mouse (93-97%; P < 0.001) and tumor volume per tumor (80-91%; P < 0.001). Silibinin also moderately inhibited tumor incidence (5-15%; P < 0.01) and delayed tumor latency period (up to 4 weeks; P < 0.01-0.001). To investigate in vivo molecular mechanisms of silibinin efficacy, tumors and uninvolved skin from tumor-bearing mice were examined immunohistochemically for proliferation, p53, apoptosis, and activated caspase-3. Silibinin treatment showed a strong decrease (P < 0.001) in proliferating cell nuclear antigen positive cells and an increase in p53-positive (P < 0.005-0.001), terminal deoxynucleotidyltransferase-mediated nick end labeling-positive (P < 0.005 0.001), and cleaved caspase-3-positive cells (P < 0.001). Western blot analysis of normal skin and tumor lysates showed that silibinin decreases the levels of cyclin-dependent kinase 2 and cyclin-dependent kinase 4 and associated cyclins A, E, and D1, together with an up-regulation of Cip1/p21, Kip1/p27, and p53. Silibinin also showed a strong phosphorylation of extracellular signal-regulated protein kinase 1/2, stress-activated protein kinase/c-JUN NH2-terminal kinase 1/2, and p38 mitogen-activated protein kinases but inhibited Akt phosphorylation and decreased survivin levels with an increase in cleaved caspase-3. Together, these results show a strong preventive efficacy of silibinin against photocarcinogenesis, which involves the inhibition of DNA synthesis, cell proliferation, and cell cycle progression and an induction of apoptosis. Furthermore, these results also identify in vivo molecular mechanisms of silibinin efficacy against photocarcinogenesis. PMID- 15342427 TI - Topographic and laminar maturation of striate cortex in early postnatal marmoset monkeys, as revealed by neurofilament immunohistochemistry. AB - The maturation of pyramidal neurons in the primary visual cortex (V1) of marmoset monkeys was investigated using an antibody (SMI-32) to non-phosphorylated neurofilament protein (NNF). Analysis of animals aged between birth and postnatal day 91 (PD 91, which corresponds approximately to the peak of synaptogenesis in this species) revealed discrete changes in both the laminar and the areal distribution of NNF. At PD 0, the upper part of layer 6 contained darkly labelled neurons and associated neuropil, including axons. In this layer a centroperipheral gradient, with more labelled cells in the foveal representation, was apparent at PD 0. This topographic gradient gradually disappeared, and by PD 91 a similar density of labelled layer 6 cells was observed throughout V1. Labelled cells were not apparent in layer 3C until PD 7, and were not distributed according to a topographic gradient. Labelled cells were first observed in layer 3B(alpha) at PD 28, when they formed a centroperipheral gradient similar to that seen in layer 6. This gradient was still evident in an adult animal. These results demonstrate an inside-out profile of postnatal cortical development, with the topographic pattern of maturation of V1 mimicking the centroperipheral gradient of maturation in the retina. PMID- 15342428 TI - Oxidative damage and defective DNA repair is linked to apoptosis of migrating neurons and progenitors during cerebral cortex development in Ku70-deficient mice. AB - DNA repair plays a critical, but imprecisely defined role in neuronal survival during cortical neurogenesis. We examined cortical development in mice deficient for the DNA end-joining protein, Ku70. At gestational day 14.5, corresponding to the peak of neurogenesis, the Ku70(-/-) embryonic cerebral cortex displayed 25- to 30-fold more cell death than heterozygous littermates, as judged by DNA breaks, pyknosis and active caspase-3. In Ku70(-/-) embryos only, large clusters of dying neurons were found in the intermediate zone. Cell death declined until P4, when the number of dying cells became comparable to that in heterozygous mice. Two groups of dying cells were evident: a GLAST(+) neural progenitor population in the subventricular and ventricular zones, and a doublecortin(+) immature neuron population in the intermediate zone, the latter exhibiting strong staining for oxidative DNA damage. Antioxidants and lower oxygen tension reduced the high levels of neuronal death in primary cortical cultures derived from Ku70( /-) mice, but not the low levels of cell death in wildtype cortical cultures. Results indicate migrating cortical neurons undergo oxidative DNA damage, which is normally repaired by non-homologous end joining. Failure to repair oxidative damage triggers a form of apoptosis involving caspase-3 activation. PMID- 15342429 TI - Larger interregional synchrony is associated with greater behavioral success in a complex sensory integration task in humans. AB - Successful behavior depends on effective communication between distant brain regions. Moreover, disturbance of effective communication can cause neurological symptoms like apraxia, dyslexia or object agnosia. Interregional communication can be assessed by coherence analysis of synchronized neuronal oscillations, and has been referred to as synchrony or "binding". The concept of synchrony as a means of information coding is attractive, but its functional relevance has been challenged. We hypothesized that if synchrony is functionally relevant in humans, then more synchrony should determine better behavioral performance. Here, we show in a visuotactile integration task that the amount of low-frequency (7-13Hz), long-range electroencephalographic coherence between visual and sensorimotor cortex is significantly correlated with the level of performance. Trials with highest coherences were the most successful ones and vice versa in the absence of differences in regional activation measured as task-related spectral power. In summary, quantitatively linking the amount of long-range synchrony with the degree of behavioral success in humans, the present data suggest that the ability to generate topographically specific synchrony of high amplitude is functionally relevant for behavioral success. They also raise the possibility that the magnitude of regional activation is less representative of the efficacy of brain functioning than interregional synchrony. PMID- 15342430 TI - A distributed left hemisphere network active during planning of everyday tool use skills. AB - Determining the relationship between mechanisms involved in action planning and/or execution is critical to understanding the neural bases of skilled behaviors, including tool use. Here we report findings from two fMRI studies of healthy, right-handed adults in which an event-related design was used to distinguish regions involved in planning (i.e. identifying, retrieving and preparing actions associated with a familiar tools' uses) versus executing tool use gestures with the dominant right (experiment 1) and non-dominant left (experiment 2) hands. For either limb, planning tool use actions activates a distributed network in the left cerebral hemisphere consisting of: (i) posterior superior temporal sulcus, along with proximal regions of the middle and superior temporal gyri; (ii) inferior frontal and ventral premotor cortices; (iii) two distinct parietal areas, one located in the anterior supramarginal gyrus (SMG) and another in posterior SMG and angular gyrus; and (iv) dorsolateral prefrontal cortex (DLFPC). With the exception of left DLFPC, adjacent and partially overlapping sub-regions of left parietal, frontal and temporal cortex are also engaged during action execution. We suggest that this left lateralized network constitutes a neural substrate for the interaction of semantic and motoric representations upon which meaningful skills depend. PMID- 15342431 TI - Activation of medial prefrontal cortex by phencyclidine is mediated via a hippocampo-prefrontal pathway. AB - Phencyclidine (PCP) is a psychotomimetic drug that elicits schizophrenia-like symptoms in healthy persons, and administration of PCP to animals is used as a pharmacological model of schizophrenia. We recently demonstrated that systemic administration of PCP to rats produces long-lasting activation of medial prefrontal cortex (mPFC) neurons with augmentation of locomotor activity, whereas direct application of PCP to mPFC neurons has little effect on their firing activity. These findings suggest that PCP-induced activation of mPFC neurons is elicited mainly via excitatory inputs from regions outside the mPFC. In the present study, we examined effects of local application of PCP to the ventral hippocampus (vHIP) on firing activity of PFC neurons in freely moving rats. PCP locally perfused into the vHIP increased spontaneous discharges of PFC neurons during perfusion with augmentation of locomotor activity. Local application of a more selective NMDA receptor antagonist, MK801, to vHIP neurons under anesthesia increased the spontaneous firing rates of most neurons directly projecting to the mPFC, whereas local application of MK801 to mPFC neurons did not induce excitatory responses in any of those neurons. The present results indicate that tonic excitatory inputs from the vHIP to the PFC may trigger development of behavioral abnormalities. PMID- 15342432 TI - Hearing lips: gamma-band activity during audiovisual speech perception. AB - Auditory pattern changes have been shown to elicit increases in magnetoencephalographic gamma-band activity (GBA) over left inferior frontal cortex, forming part of the putative auditory ventral "what" processing stream. The present study employed a McGurk-type paradigm to assess whether GBA would be associated with subjectively perceived changes even when auditory stimuli remain unchanged. Magnetoencephalograms were recorded in 16 human subjects during audiovisual mismatch perception. Both infrequent visual (auditory /ta/ + visual /pa/) and acoustic deviants (auditory/pa/ + visual /ta/) were compared with frequent audiovisual standards (auditory /ta/ and visual /ta/). Statistical probability mapping revealed spectral amplitude increases at approximately 75 and approximately 78 Hz to visual deviants. GBA to visual deviants peaked 160 ms after auditory stimulus onset over posterior parietal cortex, at 270 ms over occipital areas and at 320 ms over left inferior frontal cortex. The latter GBA enhancement was consistent with the increase observed previously to pure acoustic mismatch, supporting a role of left inferior frontal cortex for the representation of perceived auditory pattern change. The preceding gamma-band changes over posterior areas may reflect processing of incongruent lip movements in visual motion areas and back-projections to earlier visual cortex. PMID- 15342433 TI - A three-dimensional MRI atlas of the mouse brain with estimates of the average and variability. AB - Although there is growing interest in finding mouse models of human disease, no technique for quickly and quantitatively determining anatomical mutants currently exists. Magnetic resonance imaging (MRI) is ideally suited to probe fine structures in mice. This technology is three-dimensional, non-destructive and rapid compared to histopathology; hence MRI scientists have been able to create detailed three-dimensional images of 60 mum resolution or better. The data is digital which lends itself to sophisticated image processing algorithms. Here we show a variational MRI atlas constructed from nine excised brains of 8 week old 129S1/SvImJ male mice. This new type of atlas is comprised of an unbiased average brain--created from alignment of the individual brains--and the mathematical descriptors of anatomical variation across the individuals. We found that the majority of internal points in the individuals never varied more than 117 microm from equivalent points in the atlas. A three-dimensional annotation of the average image was performed and used to estimate the mean and standard deviation of volumes in a variety of structures across the individual brains; these volumes never differed by more than 5%. Our results indicate that variational atlases of inbred strains represent a well-defined basis against which mutant outliers can be readily compared. PMID- 15342434 TI - Dorsal posterior parietal rTMS affects voluntary orienting of visuospatial attention. AB - Patients with lesions in posterior parietal cortex (PPC) are relatively unimpaired in voluntarily directing visual attention to different spatial locations, while many neuroimaging studies in healthy subjects suggest dorsal PPC involvement in this function. We used an offline repetitive transcranial magnetic stimulation (rTMS) protocol to study this issue further. Ten healthy participants performed a cue-target paradigm. Cues prompted covert orienting of spatial attention under voluntary control to either a left or right visual field position. Targets were flashed subsequently at the cued or uncued location, or bilaterally. Following rTMS over right dorsal PPC, (i) the benefit for target detection at cued versus uncued positions was preserved irrespective of cueing direction (left- or rightward), but (ii) leftward cueing was associated with a global impairment in target detection, at all target locations. This reveals that leftward orienting was still possible after right dorsal PPC stimulation, albeit at an increased overall cost for target detection. In addition, rTMS (iii) impaired left, but (iv) enhanced right target detection after rightward cueing. The finding of a global drop in target detection during leftward orienting with a spared, relative detection benefit at the cued (left) location (i-ii) suggests that right dorsal PPC plays a subsidiary rather than pivotal role in voluntary spatial orienting. This finding reconciles seemingly conflicting results from patients and neuroimaging studies. The finding of attentional inhibition and enhancement occurring contra- and ipsilaterally to the stimulation site (iii-iv) supports the view that spatial attention bias can be selectively modulated through rTMS, which has proven useful to transiently reduce visual hemispatial neglect. PMID- 15342435 TI - Anteromedial temporal cortex supports fine-grained differentiation among objects. AB - Patients with damage to left anteromedial temporal cortex often show a striking deficit: they fail to recognize animals and other living things. This failure of recognition presents an important challenge to theories of the neural representation of conceptual knowledge. Here we propose that this lesion behaviour association arises because polymodal neurons in anteromedial temporal cortex integrate simple features into complex feature conjunctions, providing the neural infrastructure for differentiating among objects. PMID- 15342436 TI - Multi-item working memory--a behavioral study. AB - Macaque monkeys were trained to recognize the repetition of one of the images already seen in a sequence of random length. On average, performance decreased with sequence length. However, this was due to a complex combination of factors, as follows: performance was found to decrease with the separation in the sequence of the test (repetition image) from the cue (its first appearance in the sequence), for trials with sequences of fixed length. In contrast, performance improved as a function of sequence length, for equal cue-test separations. Reaction times followed a complementary trend: they increased with cue-test separation and decreased with sequence length. The frequency of false positives (FPs) indicates that images are not always removed from working memory between successive trials, and that the monkeys rarely confuse different images. The probability of miss errors depends on number of intervening stimulus presentations, while FPs depend on elapsed time. A simple two-state stochastic model of multi-item working memory is proposed that guides the account for the main effects of performance and false positives, as well as their interaction. In the model, images enter WM when they are presented, or by spontaneous jump-in. Misses are due to spontaneous jump-out of images previously seen. PMID- 15342437 TI - Kinematically specific interhemispheric inhibition operating in the process of generation of a voluntary movement. AB - Unilateral hand movements are accompanied by a transient decrease in corticospinal (CS) excitability of muscles in the opposite hand. However, the rules that govern this phenomenon are not completely understood. We measured the amplitude of motor evoked potentials (MEP) in the left first dorsal interosseus (FDI) elicited by transcranial magnetic stimulation (TMS) of the primary motor cortex in order to assess CS excitability changes that preceded eight possible combinations of unilateral and bilateral index finger movements with different right hand positions. Left FDI MEP amplitude (MEP(Left FDI)) increased when this muscle acted as an agonist and tended to decrease when it was an antagonist. Additionally, MEP(Left FDI) decreased substantially before right index finger abduction (a movement mediated by the right FDI) when both hands were lying flat (a movement mirroring left index finger abduction) but not when the right hand was turned at 90 degrees or flat with the palm up. Therefore, CS excitability of the resting FDI was differentially modulated depending on the direction of the opposite index finger movement, regardless of muscles engaged in the task. These results indicate that inhibitory interactions preceding unilateral finger movements are determined by movement kinematics possibly to counteract the default production of mirror motions. PMID- 15342438 TI - The neural circuitry of pre-attentive auditory change-detection: an fMRI study of pitch and duration mismatch negativity generators. AB - Electrophysiological studies have revealed a pre-attentive change-detection system in the auditory modality. This system emits a signal termed the mismatch negativity (MMN) when any detectable change in a regular pattern of auditory stimulation occurs. The precise intracranial sources underlying MMN generation, and in particular whether these vary as a function of the acoustic feature that changes, is a matter of some debate. Using functional magnetic resonance imaging, we show that anatomically distinct networks of auditory cortices are activated as a function of the deviating acoustic feature--in this case, tone frequency and tone duration--strongly supporting the hypothesis that MMN generators in auditory cortex are feature dependent. We also detail regions of the frontal and parietal cortices activated by change-detection processes. These regions also show feature dependence and we hypothesize that they reflect recruitment of attention switching mechanisms. PMID- 15342439 TI - Acquisition and performance of delayed-response tasks: a neural network model. AB - We study the time evolution of a neural network model as it learns the three stages of a visual delayed-matching-to-sample (DMS) task: identification of the sample, retention during delay, and matching of sample and target, ignoring distractors. We introduce a neurobiologically plausible, uncommitted architecture, comprising an "executive" subnetwork gating connections to and from a "working" layer. The network learns DMS by reinforcement: reward-dependent synaptic plasticity generates task-dependent behaviour. During learning, working layer cells exhibit stimulus specialization and increased tuning of their firing. The emergence of top-down activity is observed, reproducing aspects of prefrontal cortex control on activity in the visual areas of inferior temporal cortex. We observe a lability of neural systems during learning, with a tendency to encode spurious associations. Executive areas are instrumental during learning to prevent such associations; they are also fundamental for the "mature" network to keep passing DMS. In the mature model, the working layer functions as a short term memory. The mature system is remarkably robust against cell damage and its performance degrades gracefully as damage increases. The model underlines that executive systems, which regulate the flow of information between working memory and sensory areas, are required for passing tests such as DMS. At the behavioural level, the model makes testable predictions about the errors expected from subjects learning the DMS. PMID- 15342440 TI - Positive evidence against human hippocampal involvement in working memory maintenance of familiar stimuli. AB - Subjects (n = 40) performed a delayed item recognition task for visually presented letters with three set sizes (1, 3 or 6 letters). Accuracy was close to ceiling at all set sizes, so we took set size as a proxy for WM load (i.e. the amount of information being maintained in WM). Functional magnetic resonance imaging (fMRI) signal associated with the delay period increased in a nearly linear fashion with WM load in the left inferior frontal gyrus/anterior insula (possibly Broca's area, BA 44/45), right anterior insula, bilateral caudate, bilateral precentral gyrus (BA 6), bilateral middle frontal gyrus (BA 9/46), bilateral inferior parietal lobule (with foci in both BA 39 and 40), left superior parietal lobule (BA 7), medial frontal gyrus (BA 6), anterior cingulate gyrus (BA 32) and bilateral superior frontal gyrus (BA 8). These results lend support to the idea that at least some of the cortical mechanisms of WM maintenance, potentially rehearsal, exhibit a scaling with WM load. In contrast, the delay-related fMRI signal in hippocampus followed an inverted U-shape, being greatest during the intermediate level of WM load, with relatively lower values at the lowest and highest levels of WM load. This pattern of delay-related fMRI activity, orthogonal to WM load, is seemingly not consonant with a role for hippocampus in WM maintenance of phonologically codable stimuli. This finding could possibly be related more to the general familiarity of the letter stimuli than their phonological codability per se. PMID- 15342441 TI - Risk of non-Hodgkin's lymphoma and family history of lymphatic, hematologic, and other cancers. AB - BACKGROUND: An elevated risk of developing non-Hodgkin's lymphoma (NHL) has been associated with a family history of NHL and several other malignancies, but the magnitude of risks and mechanisms are uncertain. METHODS: We used self-reported family history data from a recent multicenter U.S.-based case-control studies of NHL to evaluate familial aggregation of NHL with various hematolymphoproliferative and other cancers. Estimates of familial aggregation were obtained as hazard ratios (HR) that compare incidence of different cancers in first-degree relatives of NHL cases with that in the first-degree relatives of NHL controls. Limitations of the study included low participation rates (76% for cases and 52% for controls) and potential differential accuracy of recall. RESULTS: Risk of NHL was elevated in relatives of NHL cases [HR, 2.9; 95% confidence interval (95% CI), 0.95-8.53]; the aggregation seems to be stronger for siblings (HR, 7.6; 95% CI, 0.98-58.8) and for male relatives (HR, 6.2; 95% CI, 0.77-50.0). Risk of Hodgkin's lymphoma seems to be also elevated among relatives of early-onset (<50 years) NHL cases (HR, 3.2; 95% CI, 0.88-11.6). Evaluation of family history of other cancers provided modest evidence for an increased risk of melanoma of the skin (HR, 2.9; 95% CI, 1.08-7.75), pancreatic cancer (HR, 2.1; 95% CI, 0.96-4.43), stomach cancer (HR, 1.8; 95% CI, 0.91-3.63), and prostate cancer (HR, 1.3; 95% CI, 0.87-1.99). CONCLUSIONS: These results are consistent with previous findings of familial aggregation of NHL, Hodgkin's lymphoma, and a few other cancers. The pattern of male-specific and sibling specific familial aggregation of NHL we observed, if confirmed, may shed new light on the possible mechanisms that underlie familial aggregation of the disease. PMID- 15342442 TI - Raw versus cooked vegetables and cancer risk. AB - This review of the medical literature from 1994 to 2003 summarizes the relationship between raw and cooked vegetables and cancer risk and examines whether they may affect cancer risk differently. Twenty-eight studies examined the relationship between raw and cooked vegetables and risk for various cancers. Twenty-one studies assessed raw, but not cooked, vegetables and cancer risk. The majority of these assessed risk of oral, pharyngeal, laryngeal, esophageal, lung, gastric, and colorectal cancers. Most showed that vegetables, raw or cooked, were inversely related to these cancers. However, more consistent results were found for oral, pharyngeal, laryngeal, esophageal, and gastric cancers. Nine of the 11 studies of raw and cooked vegetables showed statistically significant inverse relationships of these cancers with raw vegetables, but only 4 with cooked vegetables. The few studies of breast, lung, and colorectal cancers also suggested an inverse relationship with both raw and cooked vegetables, but these results were less consistent. In the two studies of prostate cancer, there was no association with either raw or cooked vegetables. One of two bladder cancer studies found an inverse relationship with cooked, but not raw, vegetables. Possible mechanisms by which cooking affects the relationship between vegetables and cancer risk include changes in availability of some nutrients, destruction of digestive enzymes, and alteration of the structure and digestibility of food. Both raw and cooked vegetable consumption are inversely related to epithelial cancers, particularly those of the upper gastrointestinal tract, and possibly breast cancer; however, these relationships may be stronger for raw vegetables than cooked vegetables. PMID- 15342443 TI - Associations between two common variants C677T and A1298C in the methylenetetrahydrofolate reductase gene and measures of folate metabolism and DNA stability (strand breaks, misincorporated uracil, and DNA methylation status) in human lymphocytes in vivo. AB - OBJECTIVE: Homozygosity for variants of the methylenetetrahydrofolate reductase (MTHFR) gene is associated with decreased risk for colorectal cancer. We have investigated the relationships between two variants of the MTHFR gene (C677T and A1298C) and blood folate, homocysteine, and genomic stability (strand breakage, misincorporated uracil, and global cytosine methylation in lymphocytes) in a study of 199 subjects. RESULTS: The frequencies of homozygosity for the C677T and A1298C variants of the MTHFR gene were 12.6% and 14.6%, respectively. Plasma homocysteine, folate, vitamin B12, 5-methyltetrahydrofolate, and RBC folate were determined in the C677T genotypes. Plasma folate was significantly lower (P < 0.001) in the homozygous variants (6.7 +/- 0.6 ng/mL) compared with wild-types (8.8 +/- 0.4 ng/mL) and heterozygotes (9.1 +/- 0.5 ng/mL). Homocysteine was significantly higher (P < 0.05) in homozygous variants (13.2 +/- 1.1 micromol/L) compared with homozygous subjects (10.9 +/- 0.4 micromol/L). Homozygous variants had significantly lower (P < 0.05) RBC folate (84.7 +/- 6.3 ng/mL) compared with wild-types (112.2 +/- 5.2 ng/mL) and heterozygous individuals (125.1 +/- 6.6 ng/mL). No significant difference in RBC folate was observed between wild-types and heterozygotes. The A1298C variant did not influence plasma homocysteine, folate, 5-methyltetrahydrofolate, vitamin B12, or RBC folate. Lymphocyte DNA stability biomarkers (strand breaks, misincorporated uracil, and global DNA methylation) were similar for all MTHFR C677T or A1298C variants. CONCLUSION: Data from this study do not support the hypothesis that polymorphisms in the MTHFR gene increase DNA stability by sequestering 5,10-methylenetetrahydrofolate for thymidine synthesis and reducing uracil misincorporation into DNA. PMID- 15342444 TI - Dietary and lifestyle correlates of plasma insulin-like growth factor-I (IGF-I) and IGF binding protein-3 (IGFBP-3): the multiethnic cohort. AB - High circulating concentration of insulin-like growth factor-I (IGF-I) and low circulating concentration of IGF binding protein-3 (IGFBP-3) have been associated with increased risk for breast, prostate, and colorectal cancers. Building on previous work in the Multiethnic Cohort (MEC) showing significant differences in IGF-I levels across racial/ethnic groups, we investigated which lifestyle and dietary factors are associated with levels of IGF-I and IGFBP-3 in a random sample of 1,000 MEC participants, which included Native Hawaiian, African American, Japanese, Latino, and White men and women. Crude analyses confirmed the existence of differences in protein levels with race/ethnicity, sex, age, and body size. Reproductive, physical activity, smoking, and diet variables had less consistent effects. In multivariate analyses, IGF-I levels were lower and IGFBP-3 were higher in females versus males. IGF-I and IGFBP-3 declined with increasing age in both genders. Women in the highest quartile of body mass index showed depressed IGF-I and IGFBP-3 levels; in men, height was significantly positively associated with both proteins. In women, alcohol was directly associated with IGFBP-3. Both proteins were lowest among female Latinos. IGF-I was highest among female African Americans. In men, IGFBP-3 was lowest among African Americans. Overall, although these factors were statistically significant determinants of IGF-related protein levels, they did not explain much of the variation in these levels. A positive correlation was found between IGF-I levels (ng/mL) and colon cancer incidence rates (per 100,000) within the MEC by race/ethnicity for both sexes but not for either breast or prostate cancer. PMID- 15342445 TI - Flow cytometric detection of intracellular TH1/TH2 cytokines using whole blood: validation of immunologic biomarker for use in epidemiologic studies. AB - Few biological markers of immune function have been thoroughly validated for use in epidemiologic studies that involve delayed sample processing and analysis. Here, we report our validation results for flow cytometric detection of intracellular T-helper 1/T-helper 2 (Th1/Th2) cytokines using 500 microL of whole blood obtained from children and adults. The detection of Th1/Th2 cytokine profiles by flow cytometry is a practical and mechanistically relevant assay because dysregulated cytokine production has been observed in many immune mediated disorders, including cancer. We evaluated the intraassay and intraindividual and interindividual variability and the effects of a 24- to 72 hour delayed analysis on Th1 and Th2 end points. We compared the distributions of %CD4 lymphocytes, %Th1, and %Th2 in young children (age 1 year, n = 50) and adults (age 25-52 years, n = 16). Subjects sampled monthly for up to 1 year showed minimal variation in CD4, Th1, and Th2 end points. Delayed analysis of samples (up to 24 hours) resulted in no significant differences in the expression of CD4, Th1, and Th2; however, at 48 and 72 hours, all end points differed significantly from baseline (P < 0.01). A random effects model confirmed that interindividual variability was much greater than intraindividual variability for CD4 and Th1. Compared with adults, children had marginally higher %CD4, similar %Th2, but significantly lower %Th1 (P < 0.01). These results show that flow cytometric detection of CD4, Th1, and Th2 markers using whole blood is reproducible and that these biomarkers can be effectively used in human population studies that involve transported samples, delayed processing and analysis, and limited blood volumes. PMID- 15342446 TI - Transvaginal ultrasound measurement of endometrial thickness as a biomarker for estrogen exposure. AB - OBJECTIVE: In clinical settings, transvaginal ultrasound has been used to evaluate abnormal vaginal bleeding. Because the endometrium responds to estrogens, endometrial thickness may constitute a biomarker of estrogen status in postmenopausal women. This study aimed to validate the transvaginal ultrasonographic measurement of endometrial thickness as an estrogen biomarker in asymptomatic, postmenopausal women by demonstrating an association between endometrial thickness and risk factors known to be associated with estrogen exposure. METHOD: Endometrial thickness was measured in 1,271 women ages 55 to 74 years who underwent transvaginal ultrasound screening as part of the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. A questionnaire, completed before screening, provided risk factor information, including reproductive and hormone use histories. RESULTS: Endometrial thickness measurements ranged from 1 to 32 mm (median 3.0 mm). The frequencies of thicker endometrium (> or =3.0 mm), according to body mass index (BMI) quartile, were 55.2%, 66.1%, 69.7%, and 76.7% (P < 0.0001). The frequencies of thicker endometrium were 57.8%, 58.3%, and 82.6% among never users, ex-users, and current users of hormone replacement therapy (HRT), respectively (P < 0.0001). Other factors associated with thicker endometrium included age, marital status, history of uterine fibroids, years since menopause, and history of hypertension. Statistically significant associations were not seen in analyses limited to current HRT users (n = 461). In multiple variable analysis (R2 = 0.08), current HRT use (P < 0.0001) and higher BMI (P < 0.0001) were independently associated with thicker endometrium. CONCLUSION: In postmenopausal women, factors reflecting exogenous (current HRT use) and endogenous (BMI) estrogen exposure were associated with increased endometrial thickness as measured during screening transvaginal ultrasound. Practical limitations related to screening transvaginal ultrasound include measurement variability, lack of information regarding type or dose of HRT, and problems of differentiating true endometrial thickening from unrecognized endometrial polyps or fluid accumulations. Constrained by these limitations, these results partially validate a transvaginal ultrasound measurement of endometrial thickness as a potential biomarker related to estrogen status. PMID- 15342447 TI - Vitamin D, calcium, and mammographic breast densities. AB - Vitamin D and calcium are being evaluated as potential breast cancer prevention agents. This study reports on the relation of dietary vitamin D and calcium to mammographic breast densities, one of the strongest breast cancer risk factors. Participants were women ages 40 to 60 years who had had a screening mammogram in Rhode Island and eastern Massachusetts (1989-1990). Diet was assessed by semiquantitative food frequency questionnaire, and the percentage of the breast showing densities was estimated visually by a single observer without information on subjects. Multivariate logistic regression was used to compare dietary intakes of vitamin D and calcium between women classified as having few densities (< or =30% of the breast with density, n = 287) and extensive densities (> or =70% of the breast with density, n = 256). For categories of increasing vitamin D intake (<50, 50-99, 100-199, and > or =200 IU/d), adjusted odds ratios (OR) for extensive densities were 1.00 (reference), 0.51, 0.37, and 0.24, respectively (P for trend = 0.0005). For increasing calcium intake (<500, 500-749, 750-999, and > or =1,000 mg/d), adjusted ORs were 1.00 (reference), 0.63, 0.25, and 0.24, respectively (P for trend = 0.0006). Combination of higher intakes of vitamin D and calcium (> or =100 IU/d and > or =750 mg/d, respectively) were associated with a reduction of breast densities (OR, 0.28; 95% confidence interval, 0.15 0.54) compared with those consuming <100 IU/d and <750 mg/d. Increases in vitamin D and calcium intakes were associated with decreases in breast densities, suggesting that dietary vitamin D and calcium could reduce breast cancer risk possibly through influences on breast tissue morphology. PMID- 15342448 TI - Glutathione S-transferases M1, T1, and P1 and breast cancer: a pooled analysis. AB - The glutathione S-transferase (GST) genes are involved in the metabolism of various carcinogens. Deletion polymorphisms in the genes GSTM1 and GSTT1 and a base transition polymorphism at codon 105 (Ile-->Val) in GSTP1 were investigated in relation to breast cancer risk. Tobacco smoking and reproductive factors were examined as potential effect modifiers. Individual data from seven case-control studies were pooled within the International Collaborative Study on Genetic Susceptibility to Environmental Carcinogens. To measure the effect of GSTs on breast cancer risk, odds ratios and 95% confidence intervals were computed adjusting for study center and age. The modifying effect was investigated by stratification on variables of smoking habits and reproductive history. A total of 2,048 cases with breast cancer and 1,969 controls were analyzed. The relative odds ratio (95% confidence interval) of breast cancer was 0.98 (0.86-1.12) with the GSTM1 null, 1.11 (0.87-1.41) with the GSTT1 null, 1.01 (0.79-1.28) with GSTP1 heterozygous mutants, and 0.93 (0.62-1.38) with GSTP1 homozygous mutants. Stratification by smoking or reproductive factors did not reveal a modifying effect of these variables, nor was there any association between GSTM1 and age at diagnosis of breast cancer. This is the largest study investigating susceptibility to breast cancer due to polymorphisms in the GST genes. The results conclusively show that single gene GST polymorphisms do not confer a substantial risk of breast cancer to its carriers. Furthermore, GSTs did not interact with smoking or reproductive history to modify cancer risk. PMID- 15342449 TI - Dietary intake of conjugated linoleic acids and risk of premenopausal and postmenopausal breast cancer, Western New York Exposures and Breast Cancer Study (WEB Study). AB - Specific fatty acids may have differential effects on breast cancer etiology. Animal studies have suggested that conjugated linoleic acids (CLA), a group of fatty acids found predominantly in dairy products and the meat of ruminants, have potent anticarcinogenic properties. We examined breast cancer risk and dietary CLA intake among 1,122 women with primary, incident, histologically confirmed breast cancer and 2,036 controls frequency matched to cases by age, race, and county of residence. Diet was assessed with a self-administered 104-item food frequency questionnaire and other relevant data were collected by detailed in person interviews. We examined risk with intake of total CLAs and the 9c,11t-18:2 isomer of CLA (9,11 CLA). Odds ratios and 95% confidence intervals were estimated by unconditional logistic regression, adjusting for age, the residual of fat adjusted for energy, and other breast cancer risk factors. No association was observed between intakes of total CLA or 9,11 CLA and overall risk of premenopausal or postmenopausal breast cancer. We observed little association between CLA intakes and risk of estrogen receptor (ER)-negative or ER-positive tumors, although, compared with premenopausal women in the lowest quartile of 9,11 CLA intake, those in the highest quartile had a marginally significant reduction in risk of having an ER-negative tumor (odds ratio, 0.40; 95% confidence interval, 0.16-1.01). Our findings suggest that, although CLA intake was not related to overall breast cancer risk, there may be associations with tumor biology at least among premenopausal women. PMID- 15342450 TI - Fruits, vegetables, and micronutrients in relation to breast cancer modified by menopause and hormone receptor status. AB - Whether fruit, vegetable, and antioxidant micronutrient consumption is associated with a reduction in breast cancer incidence remains unresolved. To address this issue, we analyzed data from a large population-based case-control study, with consideration given to whether the associations varied with menopausal status or with clinical characteristics of the cases' disease. Study participants completed a modified Block food frequency questionnaire, which included assessment of the frequency and portion sizes of 13 fruits and fruit juices and 16 vegetables and the use of multiple and single vitamin supplements. Statistical analyses were done on 1,463 cases and 1,500 controls. Among postmenopausal women, reduced odds ratios [OR; 95% confidence intervals (95% CI)] were noted for the highest fifth, as compared with the lowest fifth, of intake of any vegetables [0.63 (0.46-0.86); P for trend < 0.01] and leafy vegetables [0.66 (0.50-0.86); P for trend = 0.03] after controlling for age and energy intake. Adjusted ORs (95% CIs) were also decreased for postmenopausal breast cancer in relation to high intake of carotenoids, alpha-carotene, beta-carotene, lutein, and particularly lycopene [0.66 (0.48-0.90); P for trend = 0.03]. Inverse associations for fruits and vegetables were stronger for postmenopausal women with estrogen receptor (ER)+ tumors (OR, 0.65; 95% CI, 0.51-0.82) than ER- tumors (OR, 0.92; 95% CI, 0.64 1.32), but results were less consistent for micronutrients. No similarly reduced associations were observed among premenopausal women. ORs did not appreciably differ by in situ or invasive breast cancer or by whether cases had begun chemotherapy. Our results support an inverse association for fruit and vegetable intake among postmenopausal but not premenopausal breast cancer, which may be more pronounced among women with ER+ tumors. PMID- 15342451 TI - Use of DNA from human stools to detect aberrant CpG island methylation of genes implicated in colorectal cancer. AB - Hypermethylation of cytosine residues in the CpG islands of tumor suppressor genes is a key mechanism of colorectal carcinogenesis. Detection and quantification of CpG island methylation in human DNA isolated from stools might provide a novel strategy for the detection and investigation of colorectal neoplasia. To explore the feasibility of this approach, colorectal biopsies and fecal samples were obtained from 32 patients attending for colonoscopy or surgery, who were found to have adenomatous polyps, colorectal cancer, or no evidence of neoplasia. A further 18 fecal samples were obtained from healthy volunteers, with no bowel symptoms. Isolated DNA was modified with sodium bisulfite and analyzed by methylation-specific PCR and combined bisulfite restriction analysis for CpG island methylation of ESR1, MGMT, HPP1, p16(INK4a), APC, and MLH1. CpG island methylation was readily detectable in both mucosal and fecal DNA with methylation-specific PCR. Using combined bisulfite restriction analysis, it was established that, in volunteers from whom biopsies were available, the levels of methylation at two CpG sites within ESR1 assayed using fecal DNA were significantly correlated with methylation in DNA from colorectal mucosa. Thus, noninvasive techniques can be used to obtain quantitative information about the level of CpG island methylation in human colorectal mucosa. The methods described here could be applied to a much expanded range of genes and may be valuable both for screening purposes and to provide greater insight into the functional consequences of epigenetic changes in the colorectal mucosa of free-living individuals. PMID- 15342452 TI - Plasma vitamin D metabolites and risk of colorectal cancer in women. AB - OBJECTIVE: Experimental evidence suggests that 1,25-dihydroxyvitamin D and its precursor, 25-hydroxyvitamin D [25(OH)D], may aid in the prevention of colorectal cancer. We therefore examined risk in relation to plasma concentrations of these vitamin D metabolites. METHODS: In a nested case-control study among women in the Nurses' Health Study, we identified 193 colorectal cancer cases, ages 46 to 78 years, diagnosed up to 11 years after blood collection. Two controls were matched per case on year of birth and month of blood draw. Odds ratios (OR) for risk of colorectal cancer were calculated using conditional logistic regression adjusted for body mass index, physical activity, smoking, family history, use of hormone replacement therapy, aspirin use, and dietary intakes. RESULTS: We found a significant inverse linear association between plasma 25(OH)D and risk of colorectal cancer (P = 0.02). Among women in the highest quintile, the OR (95% confidence interval) was 0.53 (0.27-1.04). This inverse association remained strong when limited to women > or =60 years at blood collection (P = 0.006) but was not apparent among the younger women (P = 0.70). Benefit from higher 25(OH)D concentrations was observed for cancers at the distal colon and rectum (P = 0.02) but was not evident for those at the proximal colon (P = 0.81). In contrast to 25(OH)D, we did not observe an association between 1,25-dihydroxyvitamin D and colorectal cancer, although risk was elevated among the women in the highest quintile if they were also in the lower half of the 25(OH)D distribution (OR, 2.52; 95% confidence interval, 1.04-6.11). CONCLUSION: From these results and supporting evidence from previous studies, we conclude that higher plasma levels of 25(OH)D are associated with a lower risk of colorectal cancer in older women, particularly for cancers at the distal colon and rectum. PMID- 15342453 TI - Red meat, chicken, and fish consumption and risk of colorectal cancer. AB - BACKGROUND: Red meat and processed meat consumption have been associated with increased risk of colorectal cancer in some, but not all, relevant cohort studies. Evidence on the relationship between risk of colorectal cancer and poultry and fish consumption is inconsistent. METHODS: We conducted a prospective cohort study of 37,112 residents of Melbourne, Australia recruited from 1990 to 1994. Diet was measured with a food frequency questionnaire. We categorized the frequency of fresh red meat, processed meat, chicken, and fish consumption into approximate quartiles. Adenocarcinomas of the colon or rectum were ascertained via the Victorian Cancer Registry. RESULTS: We identified 283 colon cancers and 169 rectal cancers in an average of 9 years of follow-up. For rectal cancer, the hazard ratios [95% confidence intervals (95% CI)] in the highest quartile of consumption of fresh red meat and processed meat were 2.3 (1.2-4.2; P for trend = 0.07) and 2.0 (1.1-3.4; P for trend = 0.09), respectively. The corresponding hazard ratios (95% CIs) for colon cancer were 1.1 (0.7-1.6; P for trend = 0.9) and 1.3 (0.9-1.9; P for trend = 0.06). However, for neither type of meat was the heterogeneity between subsites significant. Chicken consumption was weakly negatively associated with colorectal cancer (hazard ratio highest quartile, 0.7; 95% CI, 0.6-1.0; P for trend = 0.03), whereas hazard ratios for fish consumption were close to unity. CONCLUSION: Consumption of fresh red meat and processed meat seemed to be associated with an increased risk of rectal cancer. Consumption of chicken and fish did not increase risk. PMID- 15342454 TI - Cytochrome P450 1B1 gene polymorphisms and postmenopausal endometrial cancer risk. AB - Estrogen unopposed by progestins is a key factor in endometrial cancer etiology. Cytochrome P450 1B1 (CYP1B1), responsible for the 4-hydroxylation of estrogen, may be important in endometrial carcinogenesis, either as a regulator of estrogen availability or as a producer of potentially genotoxic estrogen metabolites. We investigated the association of CYP1B1 genotype and endometrial cancer risk in a population-based case-control study of postmenopausal Swedish women. We used the Expectation-Maximization algorithm to estimate the haplotype frequencies in the population and calculated odds ratios and 95% confidence intervals from conditional logistic regression models. In stratified analysis, we investigated the possible effects of CYP1B1 genotype on endometrial cancer risk in subgroups defined primarily by menopausal hormone use and also by body mass index, smoking, use of combined oral contraceptives, and family history. We genotyped 689 cases and 1,549 controls for the CYP1B1 single nucleotide polymorphisms m2, m3, and m4 and estimated the haplotype frequencies among controls to 0.086, 0.291, 0.452, and 0.169 for the CYP1B1*1, CYP1B1*2, CYP1B1*3, and CYP1B1*4 alleles, respectively. We found no evidence for an overall association between CYP1B1 genotype and endometrial cancer risk, nor was there any clear indication of gene environment interaction. PMID- 15342455 TI - A case-control study of diet and the risk of ovarian cancer. AB - Epidemiologic studies have suggested that some dietary factors may play a role in the etiology of ovarian cancer, but the findings have been inconsistent. We assessed the association of ovarian cancer with dietary factors in a population based case-control study in Canada. Diet information was collected on 442 incident cases of ovarian cancer diagnosed in 1994 to 1997 and 2,135 population controls via a self-administered questionnaire. Compared with women in the lowest quartile of cholesterol intake, those in the second, third, and fourth quartiles had a multivariate adjusted odds ratio [OR; 95% confidence interval (95% CI)] of 1.12 (0.81-1.56), 1.20 (0.85-1.68), and 1.42 (1.03-1.97), respectively (P for trend = 0.031). Higher egg consumption was also associated with a nonsignificant increase in ovarian cancer risk. The ORs (95% CIs) for ovarian cancer were 0.77 (0.60-1.04) and 0.76 (0.56-0.99) among women in the highest quartile of total vegetable and cruciferous vegetable intake as compared with women in the lowest quartile. Women who took supplements of vitamin E, beta-carotene, and B-complex vitamins for > or =10 years had ORs (95% CIs) of 0.49 (0.30-0.81), 0.31 (0.11 0.91), and 0.61 (0.36-1.05), respectively. However, we did not observe an association of ovarian cancer risk with dietary fat intake, including saturated, monounsaturated, and polyunsaturated fatty acids, protein, carbohydrate, dietary fiber, fruit, dairy products, meat products, fish, chicken, grain products, nut products, baked desserts, margarine, butter, mayonnaise, and supplement of multiple vitamins, vitamin A, vitamin C, calcium, iron, zinc, and selenium. Our findings suggested that ovarian cancer risk was positively associated with higher consumption of dietary cholesterol and eggs and inversely associated with higher intake of total vegetables and cruciferous vegetables and supplementation of vitamin E, beta-carotene, and B-complex vitamins. PMID- 15342456 TI - Maternal smoking and childhood leukemia and lymphoma risk among 1,440,542 Swedish children. AB - Possible in utero effects of maternal smoking on hemopoietic cancer in the offspring have been addressed previously, although the results are inconclusive. In this investigation, we take advantage of population-based registers in Sweden to examine maternal smoking during pregnancy and childhood risk of leukemia and lymphoma. Prospective data were available from 1,440,542 Swedish children born between 1983 and 1997. Proportional hazard models were used to estimate hazard ratios (HR) and 95% confidence intervals (95% CI) controlling for potential confounders. In the study base, 750 hemopoietic cancers occurred across 11 million person-years. Incidence rates per 100,000 person-years were 4.7 for acute lymphocytic leukemia (ALL), 0.45 for acute myelogenous leukemia, and 0.76 for non Hodgkin's lymphoma. Maternal smoking was associated with a lower risk of ALL (HR, 0.73; 95% CI, 0.58-0.91). On the other hand, there was a higher risk of acute myelogenous leukemia (HR, 1.41; 95% CI, 0.74-2.67) particularly among heavy (> or =10 cigarettes per day) smokers (HR, 2.28; 95% CI, 1.05-4.94). The data also suggested a small excess risk of non-Hodgkin's lymphoma (HR, 1.25; 95% CI, 0.76 2.04). Evidence from this large cohort suggests that maternal smoking affects the risk of childhood leukemia and lymphoma in the offspring. The Swedish registries provide unique opportunities to examine this research question, with a design inherently free of selection and recall biases. The apparent protective effect with ALL needs to be explored further and in no way supports maternal smoking as beneficial, given its adverse association with common pregnancy outcomes. PMID- 15342457 TI - BRCAPAP: feasibility of clinical BRCA testing on liquid-based cervical cytology: implications for biomarker development. AB - OBJECTIVE: The study was designed to test the feasibility that lower genital tract cytology is a compatible medium for robust germ line genetic analyses. METHOD: BRCA1 and/or BRCA2 gene mutational analysis was done on DNA isolated from liquid-based cervical or vaginal cytology taken from 17 consenting women (age 29 65 years) who previously had genetic counseling followed by BRACAnalysis (Myriad Genetics, Salt Lake City, UT) blood analyses. Eleven women had known mutations in either BRCA1 or BRCA2 (cases) and six had no identified mutations (controls) on entry into the study. Anonymized cytology samples were sent to Myriad Genetics with a request for testing that was limited to the degree of genomic testing previously done on the blood samples. RESULTS: One cervicovaginal specimen from a test-positive woman had inadequate cellular content that precluded gene sequencing and therefore was excluded from this analysis. For the 16 women with adequate cytologic specimens, there was 100% concordance for BRCA mutation test results between blood and genital tract cytology (kappa = 1.0; 95% confidence interval, 0.51-1.0). CONCLUSION: We have shown the feasibility of using liquid based genital tract cytology as an alternative biospecimen to blood for germ line genetic analysis using a clinical approved assay. It needs to be emphasized that any type of testing for BRCA1 or BRCA2 mutation genotype should only be done in the setting of pretest and posttest counseling. PMID- 15342458 TI - Cyclooxygenase-2 and p53 expressions in endometrial cancer. AB - Cyclooxygenase-2 (COX-2) has been known to be related with various types of carcinoma, but we have insufficient knowledge about the association between COX-2 and endometrial cancer. Many have reported a close relationship between p53 expression and a poor prognosis in endometrial cancer, but it is unclear whether p53 is an independent prognostic factor. To clarify these uncertainties, we examined the expressions of COX-2 and p53 in endometrial cancer tissues. The study was carried on 152 endometrial cancer patients who had operation at Seoul National University Hospital. Paraffin-embedded tissue blocks were sectioned and immunostained using monoclonal anti-COX-2 and anti-p53 antibodies. Twenty-seven (17.8%) specimens stained as COX-2 positive. COX-2 positivity was more frequently observed in postmenopausal patients than in premenopausal patients (8.8% versus 25.0%; P = 0.009). However, COX-2 positivity did not show a statistically significant association with any other clinicopathologic characteristic (parity, body mass index, histotype, International Federation of Gynecology and Obstetrics stage, grade, lymph node metastasis, deep myometrial invasion, or p53 overexpression). Thirty-one (20.4%) specimens showed p53 overexpression and this was significantly correlated with an advanced stage (P = 0.001), poor differentiation (P < 0.001), lymph node metastasis (P = 0.012), and deep myometrial invasion (P < 0.001). Multivariate Cox regression analysis showed that advanced stage was an independent prognostic factor of survival, but p53 overexpression was not. COX-2 may be associated with endometrial cancer carcinogenesis during the postmenopausal period but not with tumor aggressiveness and p53 overexpression. The p53 overexpression was found to be strongly associated with endometrial cancer aggressiveness. PMID- 15342459 TI - Impact of misclassification in genotype-exposure interaction studies: example of N-acetyltransferase 2 (NAT2), smoking, and bladder cancer. AB - Errors in genotype determination can lead to bias in the estimation of genotype effects and gene-environment interactions and increases in the sample size required for molecular epidemiologic studies. We evaluated the effect of genotype misclassification on odds ratio estimates and sample size requirements for a study of NAT2 acetylation status, smoking, and bladder cancer risk. Errors in the assignment of NAT2 acetylation status by a commonly used 3-single nucleotide polymorphism (SNP) genotyping assay, compared with an 11-SNP assay, were relatively small (sensitivity of 94% and specificity of 100%) and resulted in only slight biases of the interaction parameters. However, use of the 11-SNP assay resulted in a substantial decrease in sample size needs to detect a previously reported NAT2-smoking interaction for bladder cancer: 1,121 cases instead of 1,444 cases, assuming a 1:1 case-control ratio. This example illustrates how reducing genotype misclassification can result in substantial decreases in sample size requirements and possibly substantial decreases in the cost of studies to evaluate interactions. PMID- 15342460 TI - Polymorphisms in interleukin -2, -6, and -10 are not associated with gastric cardia or esophageal cancer in a high-risk chinese population. PMID- 15342461 TI - The CCCH tandem zinc-finger protein Zfp36l2 is crucial for female fertility and early embryonic development. AB - The CCCH tandem zinc finger protein, Zfp36l2, like its better-known relative tristetraprolin (TTP), can decrease the stability of AU-rich element-containing transcripts in cell transfection studies; however, its physiological importance is unknown. We disrupted Zfp36l2 in mice, resulting in decreased expression of a truncated protein in which the N-terminal 29 amino acids had been deleted (DeltaN Zfp36l2). Mice derived from different clones of ES cells exhibited complete female infertility, despite evidence from embryo and ovary transplantation experiments that they could gestate and rear wild-type young. DeltaN-Zfp36l2 females apparently cycled and ovulated normally, and their ova could be fertilized; however, the embryos did not progress beyond the two-cell stage of development. These mice represent a specific model of disruption of the earliest stages of embryogenesis, implicating Zfp36l2, a probable mRNA-binding and destabilizing protein, in the physiological control of female fertility at the level of early embryonic development. This newly identified biological role for Zfp36l2 may have implications for maternal mRNA turnover in normal embryogenesis, and conceivably could be involved in some cases of unexplained human female infertility. PMID- 15342462 TI - Regionalisation of early head ectoderm is regulated by endoderm and prepatterns the orofacial epithelium. AB - The oral epithelium becomes regionalised proximodistally early in development, and this is reflected by the spatial expression of signalling molecules such as Fgf8 and Bmp4. This regionalisation is responsible for regulating the spatial expression of genes in the underlying mesenchyme. These genes are required for the spatial patterning of bone, cartilage orofacial development and, in mammals, teeth. The mechanism and timing of this important regionalisation during head epithelium development are not known. Using lipophilic dyes to fate map the oral epithelium in chick embryos, we show that the cells that will occupy the epithelium of the distal and the proximal mandible primordium already occupy different spatial locations in the developing head ectoderm prior to the formation of the first pharyngeal arch and neural crest migration. Moreover, the ectoderm cells fated to become proximal oral epithelium express Fgf8 and this expression requires the presence of endoderm. Thus, the first fundamental patterning process in jaw morphogenesis is controlled by the early separation of specific areas of ectoderm that are regulated by ectoderm-endoderm interactions, and does not involve neural crest cells. PMID- 15342463 TI - Mouse amnionless, which is required for primitive streak assembly, mediates cell surface localization and endocytic function of cubilin on visceral endoderm and kidney proximal tubules. AB - Impaired primitive streak assembly in the mouse amnionless (amn) mutant results in the absence of non-axial trunk mesoderm, a derivative of the middle region of the primitive streak. In addition, the epiblast of amn mutants fails to increase significantly in size after E7.0, indicating that middle primitive streak assembly is mechanistically tied to the growth of the embryo during gastrulation. Amn, a novel transmembrane protein, is expressed exclusively in an extra embryonic tissue, visceral endoderm (VE), during the early post-implantation stages. We show that Amn is also expressed in kidney proximal tubules (KPT) and intestinal epithelium, which, like the VE, are polarized epithelia specialized for resorption and secretion. To explore whether Amn participates in the development or function of KPT and intestinal epithelia and to gain insight into the function of Amn during gastrulation, we constructed Amn(-/-) ES cell<-->+/+ blastocyst chimeras. While chimeras form anatomically normal kidneys and intestine, they exhibit variable, selective proteinuria, a sign of KPT malfunction. In humans, AMN has been genetically connected to Cubilin (CUBN), a multi-ligand scavenger receptor expressed by KPT, intestine and yolk sac. Loss of CUBN, the intestinal intrinsic factor (IF)-vitamin B12 receptor, results in hereditary megaloblastic anemia (MGA1), owing to vitamin B12 malabsorption. The recent report of MGA1 families with mutations in AMN suggests that AMN functions in the same pathway as CUBN. We demonstrate that Cubn is not properly localized to the cell surface in Amn(-/-) tissues in the embryo and adult mouse, and that adult chimeras exhibit selective proteinuria of Cubn ligands. This study demonstrates that Amn is an essential component of the Cubn receptor complex in vivo and suggests that Amn/Cubn is required for endocytosis/transcytosis of one or more ligands in the VE during gastrulation to coordinate growth and patterning of the embryo. Furthermore, as AMN is apparently not required for gastrulation in humans, the developmental requirements for Amn/Cubn function may not be evolutionarily conserved, possibly reflecting differences between species in the role and organization of extra-embryonic tissues. PMID- 15342464 TI - Dysregulation of ferroportin 1 interferes with spleen organogenesis in polycythaemia mice. AB - Regulatory interferences at the iron transporter ferroportin 1 (Fpn1) cause transient defects in iron homeostasis and erythropoiesis in polycythaemia (Pcm) mutant mice. The present study identified decreased Fpn1 expression in placental syncytiotrophoblast cells at late gestation as the mechanism of neonatal iron deficiency in Pcm mutants. Tissue specificity of embryonic Fpn1 dysregulation was evident from concomitant decreases in Fpn1 mRNA and protein expression in placenta and liver, as opposed to upregulation of Fpn1 protein despite decreased transcript levels in spleen, implicating post-transcriptional regulation of Fpn1. Dysregulation of Fpn1 and decreased iron levels in Pcm mutant spleens correlated with apoptotic cell death in the stroma, resulting in a semidominant spleen regression. At 7 weeks of age, a transient increase in spleen size in Pcm heterozygotes reflected a transient erythropoietin-mediated polycythemia. Structurally, Pcm mutant spleens displayed a severe defect in red pulp formation, including disruption of the sinusoidal endothelium, as well as discrete defects in white pulp organization during postnatal development. Reduced functional competence of the Pcm mutant spleen was manifested by an impaired response to chemically induced hemolytic anemia. Thus, aberrant Fpn1 regulation and iron homeostasis interferes with development of the spleen stroma during embryogenesis, resulting in a novel defect in spleen architecture postnatally. PMID- 15342465 TI - Canonical WNT signaling promotes mammary placode development and is essential for initiation of mammary gland morphogenesis. AB - Mammary glands, like other skin appendages such as hair follicles and teeth, develop from the surface epithelium and underlying mesenchyme; however, the molecular controls of embryonic mammary development are largely unknown. We find that activation of the canonical WNT/beta-catenin signaling pathway in the embryonic mouse mammary region coincides with initiation of mammary morphogenesis, and that WNT pathway activity subsequently localizes to mammary placodes and buds. Several Wnt genes are broadly expressed in the surface epithelium at the time of mammary initiation, and expression of additional Wnt and WNT pathway genes localizes to the mammary lines and placodes as they develop. Embryos cultured in medium containing WNT3A or the WNT pathway activator lithium chloride (LiCl) display accelerated formation of expanded placodes, and LiCl induces the formation of ectopic placode-like structures that show elevated expression of the placode marker Wnt10b. Conversely, expression of the secreted WNT inhibitor Dickkopf 1 in transgenic embryo surface epithelium in vivo completely blocks mammary placode formation and prevents localized expression of all mammary placode markers tested. These data indicate that WNT signaling promotes placode development and is required for initiation of mammary gland morphogenesis. WNT signals play similar roles in hair follicle formation and thus may be broadly required for induction of skin appendage morphogenesis. PMID- 15342466 TI - Drosophila double-parked is sufficient to induce re-replication during development and is regulated by cyclin E/CDK2. AB - It is important that chromosomes are duplicated only once per cell cycle. Over replication is prevented by multiple mechanisms that block the reformation of a pre-replicative complex (pre-RC) onto origins in S and G2 phase. We have investigated the developmental regulation of Double-parked (Dup) protein, the Drosophila ortholog of Cdt1, a conserved and essential pre-RC component found in human and other organisms. We find that phosphorylation and degradation of Dup protein at G1/S requires cyclin E/CDK2. The N terminus of Dup, which contains ten potential CDK phosphorylation sites, is necessary and sufficient for Dup degradation during S phase of mitotic cycles and endocycles. Mutation of these ten phosphorylation sites, however, only partially stabilizes the protein, suggesting that multiple mechanisms ensure Dup degradation. This regulation is important because increased Dup protein is sufficient to induce profound rereplication and death of developing cells. Mis-expression has different effects on genomic replication than on developmental amplification from chorion origins. The C terminus alone has no effect on genomic replication, but it is better than full-length protein at stimulating amplification. Mutation of the Dup CDK sites increases genomic re-replication, but is dominant negative for amplification. These two results suggest that phosphorylation regulates Dup activity differently during these developmentally specific types of DNA replication. Moreover, the ability of the CDK site mutant to rapidly inhibit BrdU incorporation suggests that Dup is required for fork elongation during amplification. In the context of findings from human and other cells, our results indicate that stringent regulation of Dup protein is critical to protect genome integrity. PMID- 15342467 TI - ATX-2, the C. elegans ortholog of ataxin 2, functions in translational regulation in the germline. AB - Human ataxin 2 is a protein of unknown function that is implicated in the neurodegenerative disorder spinocerebellar ataxia type 2. We found that the C. elegans ortholog of ataxin 2, ATX-2, forms a complex with PAB-1, a cytoplasmic polyA-binding protein, and that ATX-2 is required for development of the germline. In the absence of ATX-2, proliferation of stem cells is reduced, and the germline is abnormally masculinized. These defects appear to result from inappropriate translational regulation that normally is mediated by the conserved KH-domain protein GLD-1. We find that MEX-3, a second KH-domain protein, exhibits a novel, ATX-2-dependent role in preventing inappropriate translation in the germline stem cells. Together, our results suggest that ATX-2 functions in translational regulation that is mediated by GLD-1 and MEX-3 proteins. PMID- 15342468 TI - Met and Hgf signaling controls hypaxial muscle and lateral line development in the zebrafish. AB - Somites give rise to a number of different embryonic cell types, including the precursors of skeletal muscle populations. The lateral aspect of amniote and fish somites have been shown to give rise specifically to hypaxial muscle, including the appendicular muscle that populates fins and limbs. We have investigated the morphogenetic basis for formation of specific hypaxial muscles within the zebrafish embryo and larvae. Transplantation experiments have revealed a developmentally precocious commitment of cells derived from pectoral fin level somites to forming hypaxial and specifically appendicular muscle. The fate of transplanted somites cannot be over-ridden by local inductive signals, suggesting that somitic tissue may be fixed at an early point in their developmental history to produce appendicular muscle. We further show that this restriction in competence is mirrored at the molecular level, with the exclusive expression of the receptor tyrosine kinase met within somitic regions fated to give rise to appendicular muscle. Loss-of-function experiments reveal that Met and its ligand, hepatocyte growth factor, are required for the correct morphogenesis of the hypaxial muscles in which met is expressed. Furthermore, we demonstrate a requirement for Met signaling in the process of proneuromast deposition from the posterior lateral line primordia. PMID- 15342469 TI - Identification and characterization of potent CYP3A4 inhibitors in Schisandra fruit extract. AB - Schisandra fruit, a Schisandraceae family herb, is used as a component in Kampo medicines (developed from Chinese medicines, but established in Japan). It can act as a sedative and antitussive, improve hepatic function, and give a general tonic effect. An extract of Schisandra fruit has been shown with a potent inhibitory effect on human liver microsomal erythromycin N-demethylation activity mediated by cytochrome P450 3A4 (CYP3A4). The present study was conducted to identify Schisandra fruit components having inhibitory effects on CYP3A4 by surveying the effect on human liver microsomal erythromycin N-demethylation activity. Known components of Schisandra fruit, gomisins B, C, G, and N and gamma shizandrin, showed inhibitory effects on N-demethylation activity. Among these components, gomisin C displayed the most potent and competitive inhibitory effect, with a Ki value of 0.049 microM. Furthermore, the inhibitory effect of gomisin C was stronger than that of ketoconazole (Ki = 0.070 microM), a known potent CYP3A4 inhibitor. Gomisin C, however, inhibited CYP1A2-, CYP2C9-, CYP2C19 , and CYP2D6-dependent activities only to a limited extent (IC50 values >10 microM). Moreover, gomisin C inactivated human liver microsomal erythromycin N demethylation activity in a time- and concentration-dependent manner. The inactivation kinetic parameters k(inact) and K(I) were 0.092 min(-1) and 0.399 microM, respectively. The human liver microsomal erythromycin N-demethylation activity inactivated by gomisin C did not recover on dialysis of the microsomes. Spectral scanning of CYP3A4 with gomisin C yielded an absorbance at 455 nm, suggesting that gomisin C inactivated the cytochrome P450 via the formation of a metabolite intermediate complex. This pattern is consistent with the metabolism of the methylenedioxy substituent in gomisin C. These results indicate that gomisin C is a mechanism-based inhibitor that not only competitively inhibits but irreversibly inactivates CYP3A4. PMID- 15342470 TI - Utility of recombinant enzyme kinetics in prediction of human clearance: impact of variability, CYP3A5, and CYP2C19 on CYP3A4 probe substrates. AB - A systematic kinetic analysis of the metabolism of five benzodiazepines (low to high clearance compounds) was performed in CYP3A4, CYP3A5, and CYP2C19 baculovirus-expressed recombinant systems. The data obtained in the expression systems were scaled and compared with human liver microsomal predicted clearance and observed in vivo values, using either cytochrome P450 relative activity factors (RAFs) or the relative abundance approach. Interindividual variability, both in content (CYP3A4, CYP3A5) and activity (CYP3A4, CYP3A5, and CYP2C19), were incorporated in the clearance prediction by bootstrap analysis. These resampling Monte Carlo-based simulations were performed to justify any distribution assumptions in the generated range of the predicted clearance due to a limited sample size. This approach allowed extrapolation of the recombinant clearance data to specific population groups and investigation of the role of "minor" forms like CYP3A5 and CYP2C19 in comparison to the most prolific CYP3A4. The use of quinidine 3-hydroxylation and alprazolam 1'-hydroxylation as RAF markers for CYP3A4 and CYP3A5 activity, respectively, and the incorporation of variability improved the clearance prediction of the selected benzodiazepines (apart from flunitrazepam) to within 2-fold of the in vivo value. Clearance estimates from the immunoquantified protein levels were approximately 8-fold lower in comparison to the RAF approach. The differences observed in the benzodiazepine metabolite pathway ratios between CYP3A4 and CYP3A5, particularly for 1'- to 4 hydroxymidazolam and alprazolam, provided a useful measure of interindividual differences within the CYP3A family. PMID- 15342471 TI - Compound leaves: equal to the sum of their parts? AB - The leaves of seed plants can be classified as being either simple or compound according to their shape. Two hypotheses address the homology between simple and compound leaves, which equate either individual leaflets of compound leaves with simple leaves or the entire compound leaf with a simple leaf. Here we discuss the genes that function in simple and compound leaf development, such as KNOX1 genes, including how they interact with growth hormones to link growth regulation and development to cause changes in leaf complexity. Studies of transcription factors that control leaf development, their downstream targets, and how these targets are regulated are areas of inquiry that should increase our understanding of how leaf complexity is regulated and how it evolved through time. PMID- 15342472 TI - Highly specific interactions between bHLH transcription factors and chromatin during retina development. AB - Basic helix-loop-helix (bHLH) transcription factors such as atonal homolog 5 (ATH5) and neurogenin 2 (NGN2) determine crucial events in retinogenesis. Using chromatin immunoprecipitation, we demonstrate that their interactions with target promoters undergo dynamic changes as development proceeds in the chick embryo. Chick ATH5 associates with its own promoter and with the promoter of the beta3 nicotinic receptor specifically in retinal ganglion cells and their precursors. NGN2 binds to the ATH5 promoter in retina but not in optic tectum, suggesting that interactions between bHLH factors and chromatin are highly tissue specific. The transcriptional activations of both promoters correlate with dimethylation of lysine 4 on histone H3. Inactivation of the ATH5 promoter in differentiated neurons is accompanied by replication-independent chromatin de-methylation. This report is one of the first demonstrations of correlation between gene expression, binding of transcription factors and chromatin modification in a developing neural tissue. PMID- 15342473 TI - Foxp1 regulates cardiac outflow tract, endocardial cushion morphogenesis and myocyte proliferation and maturation. AB - We have recently described a new subfamily of Fox genes, Foxp1/2/4, which are transcriptional repressors and are thought to regulate important aspects of development in several tissues, including the lung, brain, thymus and heart. Here, we show that Foxp1 is expressed in the myocardium as well as the endocardium of the developing heart. To further explore the role of Foxp1 in cardiac development, we inactivated Foxp1 through gene targeting in embryonic stem cells. Foxp1 mutant embryos have severe defects in cardiac morphogenesis, including outflow tract septation and cushion defects, a thin ventricular myocardial compact zone caused by defects in myocyte maturation and proliferation, and lack of proper ventricular septation. These defects lead to embryonic death at E14.5 and are similar to those observed in other mouse models of congenital heart disease, including Sox4 and Nfatc1 null embryos. Interestingly, expression of Sox4 in the outflow tract and cushions of Foxp1 null embryos is significantly reduced, while remodeling of the cushions is disrupted, as demonstrated by reduced apoptosis and persistent Nfatc1 expression in the cushion mesenchyme. Our results reveal a crucial role for Foxp1 in three aspects of cardiac development: (1) outflow tract development and septation, (2) tissue remodeling events required for cardiac cushion development, and (3) myocardial maturation and proliferation. PMID- 15342474 TI - Action of fat, four-jointed, dachsous and dachs in distal-to-proximal wing signaling. AB - In the Drosophila wing, distal cells signal to proximal cells to induce the expression of Wingless, but the basis for this distal-to-proximal signaling is unknown. Here, we show that three genes that act together during the establishment of tissue polarity, fat, four-jointed and dachsous, also influence the expression of Wingless in the proximal wing. fat is required cell autonomously by proximal wing cells to repress Wingless expression, and misexpression of Wingless contributes to proximal wing overgrowth in fat mutant discs. Four-jointed and Dachsous can influence Wingless expression and Fat localization non-autonomously, consistent with the suggestion that they influence signaling to Fat-expressing cells. We also identify dachs as a gene that is genetically required downstream of fat, both for its effects on imaginal disc growth and for the expression of Wingless in the proximal wing. Our observations provide important support for the emerging view that Four-jointed, Dachsous and Fat function in an intercellular signaling pathway, identify a normal role for these proteins in signaling interactions that regulate growth and patterning of the proximal wing, and identify Dachs as a candidate downstream effector of a Fat signaling pathway. PMID- 15342475 TI - kette and blown fuse interact genetically during the second fusion step of myogenesis in Drosophila. AB - Drosophila myoblast fusion proceeds in two steps. The first one gives rise to small syncytia, the muscle precursor cells, which then recruit further fusion competent myoblasts to reach the final muscle size. We have identified Kette as an essential component for myoblast fusion. In kette mutants, founder cells and fusion-competent myoblasts are determined correctly and overcome the very first fusion. But then, at the precursor cell stage, fusion is interrupted. At the ultrastructural level, fusion is characterised by cell-cell recognition, alignment, formation of prefusion complexes, electron dense plaques and membrane breakdown. In kette mutants, electron dense plaques of aberrant length accumulate and fusion is interrupted owing to a complete failure of membrane breakdown. Furthermore, we show that kette interacts genetically with blown fuse (blow) which is known to be required to proceed from prefusion complexes to the formation of the electron dense plaques. Interestingly, a surplus of Kette can replace Blow function during myogenesis. We propose a model in which Dumbfounded/Sticks and stones-dependent cell adhesion is mediated over Rolling Pebbles, Myoblast city, Crk, Blown fuse and Kette, and thus induces membrane fusion. PMID- 15342476 TI - vhnf1 integrates global RA patterning and local FGF signals to direct posterior hindbrain development in zebrafish. AB - The vertebrate hindbrain is transiently divided along the anterior-posterior axis into seven morphologically and molecularly distinct segments, or rhombomeres, that correspond to Hox expression domains. The establishment of a proper 'hox code' is required for the development of unique rhombomere identities, including specification of neuronal fates. valentino (val), the zebrafish ortholog of mafB/Kreisler (Kr), encodes a bZip transcription factor that is required cell autonomously for the development of rhombomere (r) 5 and r6 and for activation of Hox group 3 gene expression. Recent work has demonstrated that the expression of val itself depends on three factors: retinoic acid (RA) signals from the paraxial mesoderm; fibroblast growth factor (Fgf) signals from r4; and variant hepatocyte nuclear factor 1 (vhnf1, also known as tcf2), a homeodomain transcription factor expressed posterior to the r4-5 boundary. We have investigated the interactions between these inputs onto val expression in the developing zebrafish hindbrain. We show that RA induces val expression via activation of vhnf1 expression in the hindbrain. Fgf signals from r4, acting through the MapK pathway, then cooperate with Vhnf1 to activate val expression and subsequent r5 and r6 development. Additionally, vhnf1 and val function as part of a multistep process required for the repression of r4 identity in the posterior hindbrain. vhnf1 acts largely independently of val to repress the r4 'hox code' posterior to the r4-5 boundary and therefore to block acquisition of r4-specific neuronal fates in the posterior hindbrain. However, vhnf1 is not able to repress all aspects of r4 identity equivalently. val is required downstream of vhnf1 to repress r4-like cell-surface properties, as determined by an 'Eph-ephrin code', by repressing ephrin-B2a expression in r5 and r6. The different requirements for vhnf1 and val to repress hoxb1a and ephrin-B2a, respectively, demonstrate that not all aspects of an individual rhombomere's identity are regulated coordinately. PMID- 15342477 TI - Terminal tendon cell differentiation requires the glide/gcm complex. AB - Locomotion relies on stable attachment of muscle fibres to their target sites, a process that allows for muscle contraction to generate movement. Here, we show that glide/gcm and glide2/gcm2, the fly glial cell determinants, are expressed in a subpopulation of embryonic tendon cells and required for their terminal differentiation. By using loss-of-function approaches, we show that in the absence of both genes, muscle attachment to tendon cells is altered, even though the molecular cascade induced by stripe, the tendon cell determinant, is normal. Moreover, we show that glide/gcm activates a new tendon cell gene independently of stripe. Finally, we show that segment polarity genes control the epidermal expression of glide/gcm and determine, within the segment, whether it induces glial or tendon cell-specific markers. Thus, under the control of positional cues, glide/gcm triggers a new molecular pathway involved in terminal tendon cell differentiation, which allows the establishment of functional muscle attachment sites and locomotion. PMID- 15342478 TI - Specification of adaxial cell fate during maize leaf development. AB - Dorsoventral (adaxial/abaxial) polarity of the maize leaf is established in the meristem and is maintained throughout organ development to coordinate proper outgrowth and patterning of the leaf. rolled leaf1 (rld1) and leafbladeless1 (lbl1) are required for the specification of the adaxial/upper leaf surface. rld1 encodes a class III homeodomain-leucine zipper (HD-ZIPIII) protein whose adaxial expression is spatially defined by miRNA166-directed transcript cleavage on the abaxial side. The semi-dominant Rld1-Original (Rld1-O) mutation, which results from a single nucleotide substitution in the miRNA166 complementary site, leads to persistent expression of mutant transcripts on the abaxial site. This causes the adaxialization or partial reversal of leaf polarity. By contrast, recessive mutations in lbl1 cause the formation of abaxialized leaves. The lbl1 and Rld1-O mutations mutually suppress each other, indicating that these two genes act in the same genetic pathway. Adaxial and meristematic expression of rld1 is reduced in lbl1 mutants, indicating that lbl1 acts upstream of rld1 to specify adaxial fate during primordium development. However, rld1 expression in the vasculature of lbl1 is normal, suggesting that the specification of adaxial/abaxial polarity during vascular and primordia development is governed by separate but overlapping pathways. We also show that members of the maize yabby gene family are expressed on the adaxial side of incipient and developing leaf primordia. This expression pattern is unlike that observed in Arabidopsis, where YABBY expression is correlated with abaxial cell fate. The yabby expression patterns in lbl1 and Rld1 O mutants suggest that the yabby genes act downstream in the same pathway as lbl1 and rld1. Moreover, our observations suggest that maize yabby genes may direct lateral organ outgrowth rather than determine cell fate. We propose that a single genetic pathway involving lbl1, rld1 and the yabby genes integrates positional information within the SAM, and leads to adaxial/abaxial patterning and mediolateral outgrowth of the leaf. PMID- 15342479 TI - Germ cell-autonomous Wunen2 is required for germline development in Drosophila embryos. AB - In many animals, primordial germ cells (PGCs) migrate through the embryo towards the future gonad, a process guided by attractive and repulsive cues provided from surrounding somatic cells. In Drosophila, the two related lipid phosphate phosphatases (LPPs), Wunen (Wun) and Wun2, are thought to degrade extracellular substrates and to act redundantly in somatic cells to provide a repulsive environment to steer the migration of PGCs, or pole cells. Wun and Wun2 also affect the viability of pole cells, because overexpression of either one in somatic cells causes pole cell death. However, the means by which they regulate pole cell migration and survival remains elusive. We report that Wun2 has a maternal function required for the survival of pole cells during their migration to the gonad. Maternal wun2 RNA was found to be concentrated in pole cells and pole cell-specific expression of wun2 rescued the pole cell death phenotype of the maternal wun2 mutant, suggesting that wun2 activity in pole cells is required for their survival. Furthermore, we obtained genetic evidence that pole cell survival requires a proper balance of LPP activity in pole cells and somatic cells. We propose that Wun2 in pole cells competes with somatic Wun and Wun2 for a common lipid phosphate substrate, which is required by pole cells to produce their survival signal. In somatic cells, Wun and Wun2 may provide a repulsive environment for pole cell migration by depleting this extracellular substrate. PMID- 15342480 TI - GSK3 is a multifunctional regulator of Dictyostelium development. AB - Glycogen synthase kinase 3 (GSK3) is a central regulator of metazoan development and the Dictyostelium GSK3 homologue, GskA, also controls cellular differentiation. The originally derived gskA-null mutant exhibits a severe pattern formation defect. It forms very large numbers of pre-basal disc cells at the expense of the prespore population. This defect arises early during multicellular development, making it impossible to examine later functions of GskA. We report the analysis of a gskA-null mutant, generated in a different parental strain, that proceeds through development to form mature fruiting bodies. In this strain, Ax2/gskA-, early development is accelerated and slug migration greatly curtailed. In a monolayer assay of stalk cell formation, the Ax2/gskA- strain is hypersensitive to the stalk cell-inducing action of DIF-1 but largely refractory to the repressive effect exerted by extracellular cAMP. During normal development, apically situated prestalk cells express the ecmB gene just as they commit themselves to stalk cell differentiation. In the Ax2/gskA- mutant, ecmB is expressed throughout the prestalk region of the slug, suggesting that GskA forms part of the repressive signalling pathway that prevents premature commitment to stalk cell differentiation. GskA may also play an inductive developmental role, because microarray analysis identifies a large gene family, the 2C family, that require gskA for optimal expression. These observations show that GskA functions throughout Dictyostelium development, to regulate several key aspects of cellular patterning. PMID- 15342481 TI - Kruppel is a gap gene in the intermediate germband insect Oncopeltus fasciatus and is required for development of both blastoderm and germband-derived segments. AB - Segmentation in long germband insects such as Drosophila occurs essentially simultaneously across the entire body. A cascade of segmentation genes patterns the embryo along its anterior-posterior axis via subdivision of the blastoderm. This is in contrast to short and intermediate germband modes of segmentation where the anterior segments are formed during the blastoderm stage and the remaining posterior segments arise at later stages from a posterior growth zone. The biphasic character of segment generation in short and intermediate germ insects implies that different formative mechanisms may be operating in blastoderm-derived and germband-derived segments. In Drosophila, the gap gene Kruppel is required for proper formation of the central portion of the embryo. This domain of Kruppel activity in Drosophila corresponds to a region that in short and intermediate germband insects spans both blastoderm and germband derived segments. We have cloned the Kruppel homolog from the milkweed bug, Oncopeltus fasciatus (Hemiptera, Lygaeidae), an intermediate germband insect. We find that Oncopeltus Kruppel is expressed in a gap-like domain in the thorax during the blastoderm and germband stages of embryogenesis. In order to investigate the function of Kruppel in Oncopeltus segmentation, we generated knockdown phenotypes using RNAi. Loss of Kruppel activity in Oncopeltus results in a large gap phenotype, with loss of the mesothoracic through fourth abdominal segments. Additionally, we find that Kruppel is required to suppress both anterior and posterior Hox gene expression in the central portion of the germband. Our results show that Kruppel is required for both blastoderm-derived and germband-derived segments and indicate that Kruppel function is largely conserved in Oncopeltus and Drosophila despite their divergent embryogenesis. PMID- 15342482 TI - HOXA13 regulates the expression of bone morphogenetic proteins 2 and 7 to control distal limb morphogenesis. AB - In humans and mice, loss of HOXA13 function causes defects in the growth and patterning of the digits and interdigital tissues. Analysis of Hoxa13 expression reveals a pattern of localization overlapping with sites of reduced Bmp2 and Bmp7 expression in Hoxa13 mutant limbs. Biochemical analyses identified a novel series of Bmp2 and Bmp7 enhancer regions that directly interact with the HOXA13 DNA binding domain and activate gene expression in the presence of HOXA13. Immunoprecipitation of HOXA13-Bmp2 and HOXA13-Bmp7 enhancer complexes from the developing autopod confirm that endogenous HOXA13 associates with these regions. Exogenous application of BMP2 or BMP7 partially rescues the Hoxa13 mutant limb phenotype, suggesting that decreased BMP signaling contributes to the malformations present in these tissues. Together, these results provide conclusive evidence that HOXA13 regulates Bmp2 and Bmp7 expression, providing a mechanistic link between HOXA13, its target genes and the specific developmental processes affected by loss of HOXA13 function. PMID- 15342483 TI - TGFbeta superfamily signals are required for morphogenesis of the kidney mesenchyme progenitor population. AB - The TGFbeta superfamily plays diverse and essential roles in kidney development. Gdf11 and Bmp4 are essential for outgrowth and positioning of the ureteric bud, the inducer of metanephric mesenchyme. During nephrogenesis, Bmp7 is required for renewal of the mesenchyme progenitor population. Additionally, in vitro studies demonstrate inhibitory effects of BMPs and TGFbetas on collecting duct branching and growth. Here, we explore the predicted models of TGFbeta superfamily function by cell-specific inactivation of Smad4, a key mediator of TGFbeta signaling. Using a HoxB7cre transgene expressed in ureteric bud and collecting duct, we find that development of the collecting duct is Smad4 independent. By contrast, removal of Smad4 in nephrogenic mesenchyme using the Bmp7(cre/+) allele leads to disorganization of the nephrogenic mesenchyme and impairment of mesenchyme induction. Smad4-deficient metanephric mesenchyme does not display defects in inducibility in LiCl or spinal cord induction assays. However, in situ hybridization and lineage analysis of Smad4 null mesenchyme cells at E11.5 show that the nephrogenic mesenchyme does not aggregate tightly around the ureteric bud tips, but remains loosely associated, embedded within a population of cells expressing markers of both nephrogenic mesenchyme and peripheral stroma. We conclude that the failure of recruitment of nephrogenic mesenchyme leaves a primitive population of mesenchyme at the periphery of the kidney. This population is gradually depleted, and by E16.5 the periphery is composed of cells of stromal phenotype. This study uncovers a novel role for TGFbeta superfamily signaling in the recruitment and/or organization of the nephrogenic mesenchyme at early time-points of kidney development. Additionally, we present conclusive genetic lineage mapping of the collecting duct and nephrogenic mesenchyme. PMID- 15342484 TI - The hedgehog pathway is a modulator of retina regeneration. AB - The embryonic chick has the ability to regenerate its retina after it has been completely removed. Here, we provide a detailed characterization of retina regeneration in the embryonic chick at the cellular level. Retina regeneration can occur in two distinct manners. The first is via transdifferentiation, which is induced by members of the Fibroblast growth factor (Fgf) family. The second type of retinal regeneration occurs from the anterior margin of the eye, near the ciliary body (CB) and ciliary marginal zone (CMZ). We show that regeneration from the CB/CMZ is the result of proliferating stem/progenitor cells. This type of regeneration is also stimulated by Fgf2, but we show that it can be activated by Sonic hedgehog (Shh) overexpression when no ectopic Fgf2 is present. Shh stimulated activation of CB/CMZ regeneration is inhibited by the Fgf receptor (Fgfr) antagonist, PD173074. This indicates that Shh-induced regeneration acts through the Fgf signaling pathway. In addition, we show that the hedgehog (Hh) pathway plays a role in maintenance of the retina pigmented epithelium (RPE), as ectopic Shh expression inhibits transdifferentiation and Hh inhibition increases the transdifferentiation domain. Ectopic Shh expression in the regenerating retina also results in a decrease in the number of ganglion cells present and an increase in apoptosis mostly in the presumptive ganglion cell layer (GCL). However, Hh inhibition increases the number of ganglion cells but does not have an effect on cell death. Taken together, our results suggest that the hedgehog pathway is an important modulator of retina regeneration. PMID- 15342485 TI - Hypoxia affects mesoderm and enhances hemangioblast specification during early development. AB - Hypoxia Inducible Factor (HIF), consisting of HIF1alpha and ARNT (HIF1beta) subunits, activates multiple genes in response to oxygen (O(2)) deprivation. Arnt(-/-) mice exhibit substantial defects in blood cell and vessel development. We demonstrate that hypoxia accelerates the expression of Brachyury (a mesoderm specific transcription factor), BMP4 (a mesoderm-promoting growth factor) and FLK1 (a marker of hemangioblasts, the bipotential progenitor of endothelial and hematopoietic cells) in differentiating ES cell cultures. Significantly, proliferation of embryonic hemangioblasts (BL-CFCs) is regulated by hypoxia, as Arnt(+/+) ES cells generate increased numbers of FLK1(+) cells, and BL-CFCs with accelerated kinetics in response to low O(2). This response is HIF-dependent as Arnt(-/-) ES cells produce fewer FLK1(+) cells and BL-CFCs, under both normoxic and hypoxic conditions. Interestingly, this defect is rescued when Arnt(-/-) ES cells are co-cultured with Arnt(+/+) ES cells. Vegf(+/-)or Vegf(-/-) ES cells generate proper numbers of FLK1(+) cells but fewer BL-CFCs, suggesting that additional factors regulated by HIF (other than VEGF) are involved in these early events. Thus, hypoxic responses are important for the establishment of various progenitor cells, including early mesoderm and its differentiation into hemangioblasts. Together these data suggest that ineffective responses to hypoxia in Arnt(-/-) embryos abrogate proper cardiovascular development during early embryogenesis, including the pathways controlling hemangioblast differentiation. PMID- 15342486 TI - Two heads are better than one: regulation of DNA replication by hexameric helicases. PMID- 15342487 TI - SUMO and ubiquitin in the nucleus: different functions, similar mechanisms? AB - The small ubiquitin-related modifier SUMO posttranslationally modifies many proteins with roles in diverse processes including regulation of transcription, chromatin structure, and DNA repair. Similar to nonproteolytic roles of ubiquitin, SUMO modification regulates protein localization and activity. Some proteins can be modified by SUMO and ubiquitin, but with distinct functional consequences. It is possible that the effects of ubiquitination and SUMOylation are both largely due to binding of proteins bearing specific interaction domains. Both modifications are reversible, and in some cases dynamic cycles of modification may be required for activity. Studies of SUMO and ubiquitin in the nucleus are yielding new insights into regulation of gene expression, genome maintenance, and signal transduction. PMID- 15342488 TI - Segmentation in vertebrates: clock and gradient finally joined. AB - The vertebral column is derived from somites formed by segmentation of presomitic mesoderm, a fundamental process of vertebrate embryogenesis. Models on the mechanism controlling this process date back some three to four decades. Access to understanding the molecular control of somitogenesis has been gained only recently by the discovery of molecular oscillators (segmentation clock) and gradients of signaling molecules, as predicted by early models. The Notch signaling pathway is linked to the oscillator and plays a decisive role in inter- and intrasomitic boundary formation. An Fgf8 signaling gradient is involved in somite size control. And the (canonical) Wnt signaling pathway, driven by Wnt3a, appears to integrate clock and gradient in a global mechanism controlling the segmentation process. In this review, we discuss recent advances in understanding the molecular mechanism controlling somitogenesis. PMID- 15342489 TI - Regulatory circuit design and evolution using phage lambda. AB - Bistable gene regulatory circuits can adopt more than one stable epigenetic state. To understand how natural circuits have this and other systems properties, several groups have designed regulatory circuits de novo. Here we describe an alternative approach. We have modified an existing bistable circuit, that of phage lambda. With this approach, we used powerful genetic selections to identify functional circuits and selected for variants with altered behavior. The lambda circuit involves two antagonistic repressors, CI and Cro. We replaced lambda Cro with a module that included Lac repressor and several lac operators. Using a combinatorial approach, we isolated variants with different types of regulatory behavior. Several resembled wild-type lambda--they could grow lytically, could form highly stable lysogens, and carried out prophage induction. Another variant could form stable lysogens in the presence of a ligand for Lac repressor but switched to the lytic state when the ligand was removed. Several isolates evolved toward a desired behavior under selective pressure. These results strongly support the idea that complex circuits can arise during the course of evolution by a combination of simpler regulatory modules. They also underscore the advantages of modifying a natural circuit as an approach to understanding circuit design, systems behavior, and circuit evolution. PMID- 15342490 TI - Human Rif1, ortholog of a yeast telomeric protein, is regulated by ATM and 53BP1 and functions in the S-phase checkpoint. AB - We report on the function of the human ortholog of Saccharomyces cerevisiae Rif1 (Rap1-interacting factor 1). Yeast Rif1 associates with telomeres and regulates their length. In contrast, human Rif1 did not accumulate at functional telomeres, but localized to dysfunctional telomeres and to telomeric DNA clusters in ALT cells, a pattern of telomere association typical of DNA-damage-response factors. After induction of double-strand breaks (DSBs), Rif1 formed foci that colocalized with other DNA-damage-response factors. This response was strictly dependent on ATM (ataxia telangiectasia mutated) and 53BP1, but not affected by diminished function of ATR (ATM- and Rad3-related kinase), BRCA1, Chk2, Nbs1, and Mre11. Rif1 inhibition resulted in radiosensitivity and a defect in the intra-S-phase checkpoint. The S-phase checkpoint phenotype was independent of Nbs1 status, arguing that Rif1 and Nbs1 act in different pathways to inhibit DNA replication after DNA damage. These data reveal that human Rif1 contributes to the ATM mediated protection against DNA damage and point to a remarkable difference in the primary function of this protein in yeast and mammals. PMID- 15342491 TI - Attenuation of estrogen receptor alpha-mediated transcription through estrogen stimulated recruitment of a negative elongation factor. AB - Estrogen receptor alpha (ERalpha) signaling is paramount for normal mammary gland development and function and the repression of breast cancer. ERalpha function in gene regulation is mediated by a number of coactivators and corepressors, most of which are known to modify chromatin structure and/or influence the assembly of the regulatory complexes at the level of transcription initiation. Here we describe a novel mechanism of attenuating the ERalpha activity. We show that cofactor of BRCA1 (COBRA1), an integral subunit of the human negative elongation factor (NELF), directly binds to ERalpha and represses ERalpha-mediated transcription. Reduction of the endogenous NELF proteins in breast cancer cells using small interfering RNA results in elevated ERalpha-mediated transcription and enhanced cell proliferation. Chromatin immunoprecipitation reveals that recruitment of COBRA1 and the other NELF subunits to endogenous ERalpha responsive promoters is greatly stimulated upon estrogen treatment. Interestingly, COBRA1 does not affect the estrogen-dependent assembly of transcription regulatory complexes at the ERalpha-regulated promoters. Rather, it causes RNA polymerase II (RNAPII) to pause at the promoter-proximal region, which is consistent with its in vitro biochemical activity. Therefore, our in vivo work defines the first corepressor of nuclear receptors that modulates ERalpha dependent gene expression by stalling RNAPII. We suggest that this new level of regulation may be important to control the duration and magnitude of a rapid and reversible hormonal response. PMID- 15342492 TI - Association of the transcriptional corepressor TIF1beta with heterochromatin protein 1 (HP1): an essential role for progression through differentiation. AB - The transcriptional intermediary factor 1beta (TIF1beta) is a corepressor for KRAB-domain-containing zinc finger proteins and is believed to play essential roles in cell physiology by regulating chromatin organization at specific loci through association with chromatin remodeling and histone-modifying activities and recruitment of heterochromatin protein 1 (HP1) proteins. In this study, we have engineered a modified embryonal carcinoma F9 cell line (TIF1beta(HP1box/-)) expressing a mutated TIF1beta protein (TIF1beta(HP1box)) unable to interact with HP1 proteins. Phenotypic analysis of TIF1beta(HP1box/-) and TIF1beta(+/-) cells shows that TIF1beta-HP1 interaction is not required for differentiation of F9 cells into primitive endoderm-like (PrE) cells on retinoic acid (RA) treatment but is essential for further differentiation into parietal endoderm-like (PE) cells on addition of cAMP and for differentiation into visceral endoderm-like cells on treatment of vesicles with RA. Complementation experiments reveal that TIF1beta-HP1 interaction is essential only during a short window of time within early differentiating PrE cells to establish a selective transmittable competence to terminally differentiate on further cAMP inducing signal. Moreover, the expression of three endoderm-specific genes, GATA6, HNF4, and Dab2, is down regulated in TIF1beta(HP1box/-) cells compared with wild-type cells during PrE differentiation. Collectively, these data demonstrate that the interaction between TIF1beta and HP1 proteins is essential for progression through differentiation by regulating the expression of endoderm differentiation master players. PMID- 15342493 TI - Vilse, a conserved Rac/Cdc42 GAP mediating Robo repulsion in tracheal cells and axons. AB - Slit proteins steer the migration of many cell types through their binding to Robo receptors, but how Robo controls cell motility is not clear. We describe the functional analysis of vilse, a Drosophila gene required for Robo repulsion in epithelial cells and axons. Vilse defines a conserved family of RhoGAPs (Rho GTPase-activating proteins), with representatives in flies and vertebrates. The phenotypes of vilse mutants resemble the tracheal and axonal phenotypes of Slit and Robo mutants at the CNS midline. Dosage-sensitive genetic interactions between vilse, slit, and robo mutants suggest that vilse is a component of robo signaling. Moreover, overexpression of Vilse in the trachea of robo mutants ameliorates the phenotypes of robo, indicating that Vilse acts downstream of Robo to mediate midline repulsion. Vilse and its human homolog bind directly to the intracellular domains of the corresponding Robo receptors and promote the hydrolysis of RacGTP and, less efficiently, of Cdc42GTP. These results together with genetic interaction experiments with robo, vilse, and rac mutants suggest a mechanism whereby Robo repulsion is mediated by the localized inactivation of Rac through Vilse. PMID- 15342494 TI - Arabidopsis COP10 forms a complex with DDB1 and DET1 in vivo and enhances the activity of ubiquitin conjugating enzymes. AB - COP10 is a ubiquitin-conjugating enzyme variant (UEV), which is thought to act together with COP1, DET1, and the COP9 signalosome (CSN) in Arabidopsis to repress photomorphogenesis. Here, we demonstrate that COP10 interacts with ubiquitin-conjugating enzymes (E2s) in vivo, and can enhance their activity in vitro, an activity distinct from previous characterized UEVs such as MMS2 and UEV1. Furthermore, we show that COP10 forms a complex with UV-damaged DNA-binding protein 1a (DDB1a) and de-etiolated 1 (DET1), and physically interacts with COP1 and the CSN. Purified CDD (COP10, DDB1, DET1) complex also shows enhancement of E2 activity (UEA) similar to that observed with COP10 itself. Our data suggests that COP10, along with COP1 and the CSN, promotes the degradation of positive regulators of photomorphogenesis, such as the transcription factor HY5, via the ubiquitin/26S proteasome system. Thus, the CDD complex may act as a ubiquitylation-promoting factor to regulate photomorphogenesis. PMID- 15342495 TI - D. S. Falconer and Introduction to quantitative genetics. PMID- 15342496 TI - Intralocus sexual conflict can drive the evolution of genomic imprinting. AB - Genomic imprinting is a phenomenon whereby the expression of an allele differs depending upon its parent of origin. There is an increasing number of examples of this form of epigenetic inheritance across a wide range of taxa, and imprinting errors have also been implicated in several human diseases. Various hypotheses have been put forward to explain the evolution of genomic imprinting, but there is not yet a widely accepted general hypothesis for the variety of imprinting patterns observed. Here a new evolutionary hypothesis, based on intralocus sexual conflict, is proposed. This hypothesis provides a potential explanation for much of the currently available empirical data, and it also makes new predictions about patterns of genomic imprinting that are expected to evolve but that have not, as of yet, been looked for in nature. This theory also provides a potential mechanism for the resolution of intralocus sexual conflict in sexually selected traits and a novel pathway for the evolution of sexual dimorphism. PMID- 15342498 TI - The role of cis-acting sequences governing catabolite repression control of lacS expression in the archaeon Sulfolobus solfataricus. AB - The archaeon Sulfolobus solfataricus uses a catabolite repression-like system to control production of several glycoside hydrolases. To better understand this regulatory system, studies of the regulation of expression of the beta glycosidase gene (lacS) were conducted. Expression of lacS varies in response to medium composition and to mutations at an unlinked gene called car. Despite gene overlap, expression of the lacS promoter proximal gene, SSO3017, exhibited coregulation but not cotranscription with lacS. Measurements of mRNA half-life excluded differential stability as a factor in lacS regulation. Chromosomal repositioning by homologous recombination of a lacS deletion series clarified critical cis-acting sequences required for lacS regulation. lacS repositioned at amyA exhibited increased lacS expression and compromised the response to medium composition independently of lacS 5' flanking sequence composition. In contrast, regulation of lacS by the car mutation was dependent on sequences upstream of the archaeal TATA box. Expression of a promoter fusion between lacS and the car independent malA promoter integrated either at amyA or at the natural lacS locus was insensitive to the allelic state of car. In contrast, the promoter fusion retained a response to medium composition only at the lacS locus. These results indicate that car acts at the lacS promoter and that the response to medium composition involves locus-specific sequences exclusive of those present 5' to lacS or within the lacS transcription unit. PMID- 15342497 TI - Selection in context: patterns of natural selection in the glycoprotein 120 region of human immunodeficiency virus 1 within infected individuals. AB - Evolution of the HIV-1 V3 loop was monitored in 15 subjects over a period of 5 years at approximately 6-month intervals. Putative recombination was detected in many of the sequences. Evolutionary trees were estimated from the nonrecombinant viral sequences found in each individual. Selection and altered demographic regimes were detected with logit and other contingency analyses in a highly context-dependent fashion. Mutations leading to amino acid substitutions are subject to positive selection over a broad range of clinical conditions in the nonsyncytium-inducing (NSI) form, and the growth rates of the NSI strains and their level of genetic subdivision change little in going from a healthy immune system to a severely compromised immune system. In contrast, the SI form has a significant increase in growth rate as the immune system goes from healthy to compromised, particularly in those subjects who did not receive any antiviral drug therapy. This increase in SI growth rate results in a significant growth advantage of SI over NSI when the immune system is compromised. The SI strains also show more demographic subdivision when the immune system is healthy than when the immune system is compromised, and the SI form has greater demographic subdivision than NSI in subjects with healthy immune systems who also are not receiving antiviral drug therapy. Positive selection on amino-acid-changing mutations weakens and then intensifies again in the SI strains in going from healthy to compromised immune systems. These patterns are consistent with other studies that suggest that NSI strains inhibit replication of SI strains, that the V3 loop is more hidden from the immune system in the NSI form, that evolution in the V3 loop influences cell tropism and coreceptor usage, that substrate for replication of SI forms increases as the disease progresses, and that death of CD8 cells is influenced by the type of coreceptor usage typically found in SI but not in NSI strains. Finally, the transition between NSI and SI forms is associated with a burst of evolutionary change due to strong positive selection at sites other than those that define the NSI/SI phenotypes. PMID- 15342499 TI - Pervasive genomic recombination of HIV-1 in vivo. AB - Recombinants of preexisting human immunodeficiency virus type 1 (HIV-1) strains are now circulating globally. To increase our understanding of the importance of these recombinants, we assessed recombination within an individual infected from a single source by studying the linkage patterns of the auxiliary genes of HIV-1 subtype B. Maximum-likelihood phylogenetic techniques revealed evidence for recombination from topological incongruence among adjacent genes. Coalescent methods were then used to estimate the in vivo recombination rate. The estimated mean rate of 1.38 x 10(-4) recombination events/adjacent sites/generation is approximately 5.5-fold greater than the reported point mutation rate of 2.5 x 10( 5)/site/generation. Recombination was found to be frequent enough to mask evidence for purifying selection by Tajima's D test. Thus, recombination is a major evolutionary force affecting genetic variation within an HIV-1-infected individual, of the same order of magnitude as point mutational change. PMID- 15342500 TI - The high-mobility group A-type protein CarD of the bacterium Myxococcus xanthus as a transcription factor for several distinct vegetative genes. AB - CarD is the only reported prokaryotic protein showing structural and functional features typical of eukaryotic high-mobility group A transcription factors. In prokaryotes, proteins similar to CarD appear to be confined primarily to myxobacteria. In Myxococcus xanthus, CarD has been previously shown to act as a positive element in two different regulatory networks: one for light-induced synthesis of carotenoids and the other for starvation-induced fruiting body formation. We have now tested the effect of a loss-of-function mutation in the carD gene (carD1) on the expression of a random collection of lacZ-tagged genes, which are normally expressed in the dark during vegetative growth in rich medium. Our results indicate that CarD plays a significant role in the transcriptional regulation of various indicated genes. The carD1 mutation downregulates some genes and upregulates others. Also reported here is the isolation of several mutations that suppress the strong effect of carD1 on the expression of a particular vegetative gene. One of them (sud-2) also suppresses the effect of carD1 on other vegetative genes and on fruiting-body formation. Thus, CarD and the sud-2 gene product appear to participate in a single mechanism, which underlies various apparently diverse regulatory phenomena ascribed to CarD. PMID- 15342501 TI - CDC7/DBF4 functions in the translesion synthesis branch of the RAD6 epistasis group in Saccharomyces cerevisiae. AB - CDC7 and DBF4 encode the essential Cdc7-Dbf4 protein kinase required for DNA replication in eukaryotes from yeast to human. Cdc7-Dbf4 is also required for DNA damage-induced mutagenesis, one of several postreplicational DNA damage tolerance mechanisms mediated by the RAD6 epistasis group. Several genes have been determined to function in separate branches within this group, including RAD5, REV3/REV7 (Pol zeta), RAD30 (Pol eta), and POL30 (PCNA). An extensive genetic analysis of the interactions between CDC7 and REV3, RAD30, RAD5, or POL30 in response to DNA damage was done to determine its role in the RAD6 pathway. CDC7, RAD5, POL30, and RAD30 were found to constitute four separate branches of the RAD6 epistasis group in response to UV and MMS exposure. CDC7 is also shown to function separately from REV3 in response to MMS. However, they belong in the same pathway in response to UV. We propose that the Cdc7-Dbf4 kinase associates with components of the translesion synthesis pathway and that this interaction is dependent upon the type of DNA damage. Finally, activation of the DNA damage checkpoint and the resulting cell cycle delay is intact in cdc7Delta mcm5-bob1 cells, suggesting a direct role for CDC7 in DNA repair/damage tolerance. PMID- 15342502 TI - Expansion and contraction of the DUP240 multigene family in Saccharomyces cerevisiae populations. AB - The influence of duplicated sequences on chromosomal stability is poorly understood. To characterize chromosomal rearrangements involving duplicated sequences, we compared the organization of tandem repeats of the DUP240 gene family in 15 Saccharomyces cerevisiae strains of various origins. The DUP240 gene family consists of 10 members of unknown function in the reference strain S288C. Five DUP240 paralogs on chromosome I and two on chromosome VII are arranged as tandem repeats that are highly polymorphic in copy number and sequence. We characterized DNA sequences that are likely involved in homologous or nonhomologous recombination events and are responsible for intra- and interchromosomal rearrangements that cause the creation and disappearance of DUP240 paralogs. The tandemly repeated DUP240 genes seem to be privileged sites of gene birth and death. PMID- 15342503 TI - Cyclin B-cdk activity stimulates meiotic rereplication in budding yeast. AB - Haploidization of gametes during meiosis requires a single round of premeiotic DNA replication (meiS) followed by two successive nuclear divisions. This study demonstrates that ectopic activation of cyclin B/cyclin-dependent kinase in budding yeast recruits up to 30% of meiotic cells to execute one to three additional rounds of meiS. Rereplication occurs prior to the meiotic nuclear divisions, indicating that this process is different from the postmeiotic mitoses observed in other fungi. The cells with overreplicated DNA produced asci containing up to 20 spores that were viable and haploid and demonstrated Mendelian marker segregation. Genetic tests indicated that these cells executed the meiosis I reductional division and possessed a spindle checkpoint. Finally, interfering with normal synaptonemal complex formation or recombination increased the efficiency of rereplication. These studies indicate that the block to rereplication is very different in meiotic and mitotic cells and suggest a negative role for the recombination machinery in allowing rereplication. Moreover, the production of haploids, regardless of the genome content, suggests that the cell counts replication cycles, not chromosomes, in determining the number of nuclear divisions to execute. PMID- 15342504 TI - The Aspergillus nidulans npkA gene encodes a Cdc2-related kinase that genetically interacts with the UvsBATR kinase. AB - The DNA damage response is a protective mechanism that ensures the maintenance of genomic integrity. We have used Aspergillus nidulans as a model system to characterize the DNA damage response caused by the antitopoisomerase I drug, camptothecin. We report the molecular characterization of a p34Cdc2-related gene, npkA, from A. nidulans. The npkA gene is transcriptionally induced by camptothecin and other DNA-damaging agents, and its induction in the presence of camptothecin is dependent on the uvsBATR gene. There were no growth defects, changes in developmental patterns, increased sensitivity to DNA-damaging agents, or effects on septation or growth rate in the A. nidulans npkA deletion strain. However, the DeltanpkA mutation can partially suppress HU sensitivity caused by the DeltauvsBATR and uvsD153ATRIP checkpoint mutations. We demonstrated that the A. nidulans uvsBATR gene is involved in DNA replication and the intra-S-phase checkpoints and that the DeltanpkA mutation can suppress its intra-S-phase checkpoint deficiency. There is a defect in both the intra-S-phase and DNA replication checkpoints due to the npkA inactivation when DNA replication is slowed at 6 mm HU. Our results suggest that the npkA gene plays a role in cell cycle progression during S-phase as well as in a DNA damage signal transduction pathway in A. nidulans. PMID- 15342505 TI - High-density genetic linkage maps of Phytophthora infestans reveal trisomic progeny and chromosomal rearrangements. AB - Detailed analysis of the inheritance of molecular markers was performed in the oomycete plant pathogen Phytophthora infestans. Linkage analysis in the sexual progeny of two Dutch field isolates (cross 71) resulted in a high-density map containing 508 markers on 13 major and 10 minor linkage groups. The map showed strong clustering of markers, particularly of markers originating from one parent, and dissimilarity between the parental isolates on linkage group III in the vicinity of the mating-type locus, indicating a chromosomal translocation. A second genetic map, constructed by linkage analysis in sexual progeny of two Mexican isolates (cross 68), contained 363 markers and is thus less dense than the cross 71 map. For some linkage groups the two independent linkage maps could be aligned, but sometimes markers appeared to be in a different order, or not linked at all, indicating chromosomal rearrangements between genotypes. Graphical genotyping showed that some progeny contained three copies of a homologous linkage group. This trisomy was found for several linkage groups in both crosses. Together, these analyses suggest a genome with a high degree of flexibility, which may have implications for evolution of new races and resistance development to crop protection agents. PMID- 15342506 TI - Dikaryons of the basidiomycete fungus Schizophyllum commune: evolution in long term culture. AB - The impact of ploidy on adaptation is a central issue in evolutionary biology. While many eukaryotic organisms exist as diploids, with two sets of gametic genomes residing in the same nucleus, most basidiomycete fungi exist as dikaryons in which the two genomes exist in separate nuclei that are physically paired and that divide in a coordinated manner during hyphal extension. To determine if haploid monokaryotic and dikaryotic mycelia adapt to novel environments under natural selection, we serially transferred replicate populations of each ploidy state on minimal medium for 18 months (approximately 13,000 generations). Dikaryotic mycelia responded to selection with increases in growth rate, while haploid monokaryotic mycelia did not. To determine if the haploid components of the dikaryon adapt reciprocally to one another's presence over time, we recovered the intact haploid components of dikaryotic mycelia at different time points (without meiosis) and mated them with nuclei of different evolutionary histories. We found evidence for coadaptation between nuclei in one dikaryotic line, in which a dominant deleterious mutation in one nucleus was followed by a compensatory mutation in the other nucleus; the mutant nuclei that evolved together had the best overall fitness. In other lines, nuclei had equal or higher fitness when paired with nuclei of other histories, indicating a heterozygote advantage. To determine if genetic exchange occurs between the two nuclei of a dikaryon, we developed a 24-locus genotyping system based on single nucleotide polymorphisms to monitor somatic exchange. We observed genetic exchange and recombination between the nuclei of several different dikaryons, resulting in genotypic variation in these mitotic cell lineages. PMID- 15342507 TI - A network of stimulatory and inhibitory Galpha-subunits regulates olfaction in Caenorhabditis elegans. AB - The two pairs of sensory neurons of C. elegans, AWA and AWC, that mediate odorant attraction, express six Galpha-subunits, suggesting that olfaction is regulated by a complex signaling network. Here, we describe the cellular localization and functions of the six olfactory Galpha-subunits: GPA-2, GPA-3, GPA-5, GPA-6, GPA 13, and ODR-3. All except GPA-6 localize to sensory cilia, suggesting a direct role in sensory transduction. GPA-2, GPA-3, GPA-5, and GPA-6 are also present in cell bodies and axons and GPA-5 specifically localizes to synaptic sites. Analysis of animals with single- to sixfold loss-of-function mutations shows that olfaction involves a balance between multiple stimulatory and inhibitory signals. ODR-3 constitutes the main stimulatory signal and is sufficient for the detection of odorants. GPA-3 forms a second stimulatory signal in the AWA and AWC neurons, also sufficient for odorant detection. In AWA, signaling is suppressed by GPA-5. In AWC, GPA-2 and GPA-13 negatively and positively regulate signaling, respectively. Finally, we show that only ODR-3 plays a role in cilia morphogenesis. Defects in this process are, however, independent of olfactory behavior. Our findings reveal the existence of a complex signaling network that controls odorant detection by C. elegans. PMID- 15342508 TI - Genetic suppression of intronic +1G mutations by compensatory U1 snRNA changes in Caenorhabditis elegans. AB - Mutations to the canonical +1G of introns, which are commonly found in many human inherited disease alleles, invariably result in aberrant splicing. Here we report genetic findings in C. elegans that aberrant splicing due to +1G mutations can be suppressed by U1 snRNA mutations. An intronic +1G-to-U mutation, e936, in the C. elegans unc-73 gene causes aberrant splicing and loss of gene function. We previously showed that mutation of the sup-39 gene promotes splicing at the mutant splice donor in e936 mutants. We demonstrate here that sup-39 is a U1 snRNA gene; suppressor mutations in sup-39 are compensatory substitutions in the 5' end, which enhance recognition of the mutant splice donor. sup-6(st19) is an allele-specific suppressor of unc-13(e309), which contains an intronic +1G-to-A transition. The e309 mutation activates a cryptic splice site, and sup-6(st19) restores splicing to the mutant splice donor. sup-6 also encodes a U1 snRNA and the mutant contains a compensatory substitution at its 5' end. This is the first demonstration that U1 snRNAs can act to suppress the effects of mutations to the invariant +1G of introns. These findings are suggestive of a potential treatment of certain alleles of inherited human genetic diseases. PMID- 15342509 TI - Identification of residues of the Caenorhabditis elegans LIN-1 ETS domain that are necessary for DNA binding and regulation of vulval cell fates. AB - LIN-1 is an ETS domain protein. A receptor tyrosine kinase/Ras/mitogen-activated protein kinase signaling pathway regulates LIN-1 in the P6.p cell to induce the primary vulval cell fate during Caenorhabditis elegans development. We identified 23 lin-1 loss-of-function mutations by conducting several genetic screens. We characterized the molecular lesions in these lin-1 alleles and in several previously identified lin-1 alleles. Nine missense mutations and 10 nonsense mutations were identified. All of these lin-1 missense mutations affect highly conserved residues in the ETS domain. These missense mutations can be arranged in an allelic series; the strongest mutations eliminate most or all lin-1 functions, and the weakest mutation partially reduces lin-1 function. An electrophoretic mobility shift assay was used to demonstrate that purified LIN-1 protein has sequence-specific DNA-binding activity that required the core sequence GGAA. LIN 1 mutant proteins containing the missense substitutions had dramatically reduced DNA binding. These experiments identify eight highly conserved residues of the ETS domain that are necessary for DNA binding. The identification of multiple mutations that reduce the function of lin-1 as an inhibitor of the primary vulval cell fate and also reduce DNA binding suggest that DNA binding is essential for LIN-1 function in an animal. PMID- 15342510 TI - Translating available food into the number of eggs laid by Drosophila melanogaster. AB - In Drosophila and other insects egg production is related to the nutrients available. Somehow the nutritional status of the environment is translated into hormonal signs that can be "read" by each individual egg chamber, influencing the decision to either develop into an egg or die. We have shown that BR-C is a control gene during oogenesis and that the differential expression of BR-C isoforms plays a key role in controlling whether the fate of the egg chamber is to develop or undergo apoptosis. PMID- 15342511 TI - Dynamical analysis of regulatory interactions in the gap gene system of Drosophila melanogaster. AB - Genetic studies have revealed that segment determination in Drosophila melanogaster is based on hierarchical regulatory interactions among maternal coordinate and zygotic segmentation genes. The gap gene system constitutes the most upstream zygotic layer of this regulatory hierarchy, responsible for the initial interpretation of positional information encoded by maternal gradients. We present a detailed analysis of regulatory interactions involved in gap gene regulation based on gap gene circuits, which are mathematical gene network models used to infer regulatory interactions from quantitative gene expression data. Our models reproduce gap gene expression at high accuracy and temporal resolution. Regulatory interactions found in gap gene circuits provide consistent and sufficient mechanisms for gap gene expression, which largely agree with mechanisms previously inferred from qualitative studies of mutant gene expression patterns. Our models predict activation of Kr by Cad and clarify several other regulatory interactions. Our analysis suggests a central role for repressive feedback loops between complementary gap genes. We observe that repressive interactions among overlapping gap genes show anteroposterior asymmetry with posterior dominance. Finally, our models suggest a correlation between timing of gap domain boundary formation and regulatory contributions from the terminal maternal system. PMID- 15342512 TI - Enhancer choice in cis and in trans in Drosophila melanogaster: role of the promoter. AB - Eukaryotic enhancers act over very long distances, yet still show remarkable specificity for their own promoter. To better understand mechanisms underlying this enhancer-promoter specificity, we used transvection to analyze enhancer choice between two promoters, one located in cis to the enhancer and the other in trans to the enhancer, at the yellow gene of Drosophila melanogaster. Previously, we demonstrated that enhancers at yellow prefer to act on the cis-linked promoter, but that mutation of core promoter elements in the cis-linked promoter releases enhancers to act in trans. Here, we address the mechanism by which these elements affect enhancer choice. We consider and explicitly test three models that are based on promoter competency, promoter pairing, and promoter identity. Through targeted gene replacement of the endogenous yellow gene, we show that competency of the cis-linked promoter is a key parameter in the cis-trans choice of an enhancer. In fact, complete replacement of the yellow promoter with both TATA-containing and TATA-less heterologous promoters maintains enhancer action in cis. PMID- 15342513 TI - A single-amino-acid change of the gustatory receptor gene, Gr5a, has a major effect on trehalose sensitivity in a natural population of Drosophila melanogaster. AB - Variation in trehalose sensitivity and nucleotide sequence polymorphism of the Gr5a gene encoding the gustatory receptor to sugar trehalose were investigated in 152 male lines of Drosophila melanogaster collected from a natural population. Among the observed 59 segregating sites, some pairs of sites showed significant linkage disequilibrium. A single SNP, which results in the Ala218Thr amino acid change, was significantly associated with trehalose sensitivity, as previously suggested. Threonine at amino acid position 218 was found to be the ancestral form in D. melanogaster, suggesting that low trehalose sensitivity was an ancestral form with respect to the receptor function. There was large genetic variation in trehalose sensitivity. It was continuously distributed, indicating that trehalose sensitivity measured by the behavioral assay is a quantitative trait. These results suggest that apart from the Gr5a gene, other genetic factors contribute to variation in trehalose sensitivity. Nucleotide diversity (pi) and nucleotide variation (theta) per site were 0.00874 and 0.00590, respectively. Fu and Li's test and the MK test showed no significant departure from the expectation of selective neutrality in the Gr5a gene. However, we rejected selective neutrality by Tajima's test and Fay and Wu's test with the observed level of recombination. We discuss possible causes of the observed pattern of nucleotide variation in the gustatory receptor Gr5a gene. PMID- 15342515 TI - The genetic architecture of the behavioral ontogeny of foraging in honeybee workers. AB - The initiation of foraging during the life course of honeybee workers is of central interest to understanding the division of labor in social insects, a central theme in sociobiology and behavioral research. It also provides one of the most complex phenotypic traits in biological systems because of the interaction of various external, social, and individual factors. This study reports on a comprehensive investigation of the genetic architecture of the age of foraging initiation in honeybees. It comprises an estimation of genetic variation, the study of candidate loci, and two complementary quantitative trait loci (QTL) maps using two selected, continually bred lines of honeybees. We conclude that considerable genetic variation exists between the selected lines for this central life history component. The study reveals direct pleiotropic and epistatic effects of candidate loci (including previously identified QTL for foraging behavior). Furthermore, two maps of the honeybee genome were constructed from over 400 AFLP markers. Both maps confirm the extraordinary recombinational size of the honeybee genome. On the basis of these maps, we report four new significant QTL and two more suggestive QTL that influence the initiation of foraging. PMID- 15342516 TI - Conditional expression in the malaria mosquito Anopheles stephensi with Tet-On and Tet-Off systems. AB - We report successful conditional gene expression in the malaria vector, Anopheles stephensi, on the basis of binary systems consisting of gene driver and responder transgenic lines generated by Minos-mediated germline transformation. An A. gambiae tissue-specific enhancer derived from a serpin (SRPN10) gene was utilized to control the temporal and spatial expression of doxycycline (dox)-sensitive transcriptional regulators in the driver lines. The "Tet-Off" driver utilized the tetracycline-controlled transcriptional activator (tTA) that is unable to bind and activate transcription from tetracycline operators (TetO) in the presence of dox; the "Tet-on" driver utilized the reverse tTA (rtTA) that, conversely, binds and activates TetO operators in the presence of dox. The responder lines carried insertions encompassing a LacZ reporter gene, cis-regulated by a TetO-P-element hybrid promoter. The progeny of crosses between driver and responder lines expressed beta-galactosidase under dual, tissue-specific and dox-mediated regulation. In adult rtTA/TetOPlacZ progeny, dox treatment rapidly induced beta galactosidase activity throughout the midgut epithelium and especially in malaria parasite-invaded epithelial cells. Transactivator-dependent, dox-mediated regulation was observed in hemocytes and pericardial cells using both systems. Conditional tissue-specific regulation is a powerful tool for analyzing gene function in mosquitoes and potentially for development of strategies to control disease transmission. PMID- 15342514 TI - Detecting the footprint of positive selection in a european population of Drosophila melanogaster: multilocus pattern of variation and distance to coding regions. AB - The effects on nucleotide variation of adaptations to temperate habitats and of the possible bottleneck associated with the origin of European populations of Drosophila melanogaster should be detectable in DNA sequences given the short time elapsed relative to the species population size. We surveyed nucleotide variation in 109 fragments distributed across the X chromosome in a European population of D. melanogaster to detect the footprint of positive selection. Fragments were located primarily in large noncoding regions. Multilocus tests based on Tajima's D statistic revealed a significant departure from neutral expectations in a stationary panmictic population, with an important contribution from both positive and negative D values. A positive relationship between Tajima's D values and distance to coding region was detected, with a comparative excess of significantly negative D values in the subset of fragments closer to coding regions. Also, there was a significant heterogeneity in the polymorphism to divergence ratio, with 12 fragments contributing 42% to the test statistic. Moreover, these fragments were comparatively closer to coding regions. These findings would imply positive selection events, and thus selective sweeps, during the species expansion to Europe. PMID- 15342517 TI - Extensive sex-specific nonadditivity of gene expression in Drosophila melanogaster. AB - Assessment of the degree to which gene expression is additive and heritable has important implications for understanding the maintenance of variation, adaptation, phenotypic divergence, and the mapping of genotype onto phenotype. We used whole-genome transcript profiling using Agilent long-oligonucleotide microarrays representing 12,017 genes to demonstrate that gene transcription is pervasively nonadditive in Drosophila melanogaster. Comparison of adults of two isogenic lines and their reciprocal F1 hybrids revealed 5820 genes as significantly different between at least two of the four genotypes in either males or females or across both sexes. Strikingly, while 25% of all genes differ between the two parents, 33% differ between both F1's and the parents, averaged across sexes. However, only 5% of genes show overdominance, suggesting that heterosis for expression is rare. PMID- 15342518 TI - A piggyBac transposon gene trap for the analysis of gene expression and function in Drosophila. AB - P-element-based gene and enhancer trap strategies have provided a wealth of information on the expression and function of genes in Drosophila melanogaster. Here we present a new vector that utilizes the simple insertion requirements of the piggyBac transposon, coupled to a splice acceptor (SA) site fused to the sequence encoding enhanced green fluorescent protein (EGFP) and a transcriptional terminator. Mobilization of the piggyBac splice site gene trap vector (PBss) was accomplished by heat-shock-induced expression of piggyBac transposase (PBase). We show that insertion of PBss into genes leads to fusions between the gene's mRNA and the PBss-encoded EGFP transcripts. As heterozygotes, these fusions report the normal pattern of expression of the trapped gene. As homozygotes, these fusions can inactivate the gene and lead to lethality. Molecular characterization of PBss insertion events shows that they are single copy, that they always occur at TTAA sequences, and that splicing utilizes the engineered splice site in PBss. In those instances where protein-EGFP fusions are predicted to occur, the subcellular localization of the wild-type protein can be inferred from the localization of the EGFP fusion protein. These experiments highlight the utility of the PBss system for expanding the functional genomics tools that are available in Drosophila. PMID- 15342519 TI - A genomic basis for the evolution of vertebrate transcription factors containing amino Acid runs. AB - We have previously shown that polyAla (A) tract-containing proteins frequently present runs of glycine (G), proline (P), and histidine (H) and that, in their ORFs, GC content at all codon positions is higher than that in the rest of the genome. In this study, we present new analyses of these human proteins/ORFs. We detected striking differences in codon usage for A, G, and P in and out of runs. After dividing the ORFs, we found that 5' halves were richer in runs than 3' halves. Afterward, when removing the runs, we observed that the run-rich halves (grouped irrespectively of their 5' or 3' position) had a marked statistical tendency to have more homo- and hetero-dicodons for A, G, P, and H than the run poor halves. This suggests that, in addition to the necessary GC-rich genomic background, a specific codon organization is probably required to generate these coding repeats. Homo-dicodons may indeed provide primers for run formation through polymerase slippage. The compositional analysis of human HOX genes, the most polyAla-rich family, and their comparison with their zebrafish homologs, support these hypotheses and suggest possible effects of genomic environment on ORF evolution and organismal diversification. PMID- 15342520 TI - Phenotypic variation resulting from a deficiency of epidermal growth factor receptor in mice is caused by extensive genetic heterogeneity that can be genetically and molecularly partitioned. AB - The timing of lethality caused by homozygosity for a null allele of the epidermal growth factor receptor (Egfrtm1Mag) in mice is strongly dependent on genetic background. Initial attempts to genetically map background modifiers using Swiss derived, outbred CD-1 mice were unsuccessful. To investigate the genetic architecture contributing to survival of Egfrtm1Mag homozygous embryos, the genetic variability segregating within the outbred population was partitioned by surveying viability of Egfrtm1Mag mutants using intercrosses between 129S6/SvEvTAC-Egfrtm1Mag and nine Swiss-derived, inbred strains: ALR/LtJ, ALS/LtJ, APN, APS, ICR/HaRos, NOD/LtJ, NON/LtJ, SJL/J, and SWR/J. The observations showed that these strains support varying levels of survival of Egfrtm1Mag homozygous embryos, suggesting that genetic heterogeneity within the CD-1 stock contributed to the original lack of Egfrtm1Mag modifier detection. Similar to the Swiss-derived intercrosses, nine congenic strains, derived from 129S6/SvEvTAC, AKR/J, APN, BALB/cJ, BTBR-T+ tf/tf, C3H/HeJ, C57BL/6J, DBA/2J, and FVB/NJ inbred backgrounds, also supported varying levels of survival of Egfrtm1Mag mutants. By intercrossing the congenic lines to create hybrid F1 embryos, different genetic backgrounds were found to have complementary modifiers. Analysis of the congenic lines argues against heterosis of outbred backgrounds contributing to Egfrtm1Mag phenotypic variability. A detailed analysis of the crosses suggests that modifiers function at three distinct stages of development. One class of modifiers supports survival of Egfrtm1Mag homozygous embryos to mid-gestation, another class supports development through the mid gestation transition from yolk-sac to placental-derived nutrient sources, and a third class supports survival through later stages of gestation. Data from microarray analysis using RNA from wild-type and Egfrtm1Mag mutant placentas support the existence of extensive genetic heterogeneity and suggest that it can be molecularly partitioned. This method should be generally useful to partition heterogeneity contributing to other complex traits. PMID- 15342521 TI - A genetic mechanism implicates chromosome 11 in schizophrenia and bipolar diseases. AB - The causes of schizophrenia and bipolar human psychiatric disorders are unknown. A novel somatic cell genetic model postulated nonrandom segregation of "Watson" vs. "Crick" DNA chains of both copies of a chromosome to specific daughter cells. Such an oriented asymmetric cell division causes development of healthy, functionally nonequivalent brain hemispheres. Genetic translocations of the chromosome may cause disease by disrupting the biased strand-segregation process. Only one-half of chromosome 1 and 11 translocation carriers developing disease were recently explained as a result consistent with the model (Klar 2002). Is chromosome 1 or 11 involved? Does the translocation breakpoint cause disease? Remarkably, two other unrelated chromosome 11 translocations discovered from the literature likewise caused disease in approximately 50% of carriers. Together, their breakpoints lie at three distinct regions spanning approximately 40% of chromosome 11. Thus, chromosome 11 is implicated but the breakpoints themselves are unlikely to cause the disease. The results suggest that the genetically caused disease develops without a Mendelian gene mutation. PMID- 15342522 TI - Heterogeneous patterns of variation among multiple human x-linked Loci: the possible role of diversity-reducing selection in non-africans. AB - Studies of human DNA sequence polymorphism reveal a range of diversity patterns throughout the genome. This variation among loci may be due to natural selection, demographic influences, and/or different sampling strategies. Here we build on a continuing study of noncoding regions on the X chromosome in a panel of 41 globally sampled humans representing African and non-African populations by examining patterns of DNA sequence variation at four loci (APXL, AMELX, TNFSF5, and RRM2P4) and comparing these patterns with those previously reported at six loci in the same panel of 41 individuals. We also include comparisons with patterns of noncoding variation seen at five additional X-linked loci that were sequenced in similar global panels. We find that, while almost all loci show a reduction in non-African diversity, the magnitude of the reduction varies substantially across loci. The large observed variance in non-African levels of diversity results in the rejection of a neutral model of molecular evolution with a multi-locus HKA test under both a constant size and a bottleneck model. In non Africans, some loci harbor an excess of rare mutations over neutral equilibrium predictions, while other loci show no such deviation in the distribution of mutation frequencies. We also observe a positive relationship between recombination rate and frequency spectra in our non-African, but not in our African, sample. These results indicate that a simple out-of-Africa bottleneck model is not sufficient to explain the observed patterns of sequence variation and that diversity-reducing selection acting at a subset of loci and/or a more complex neutral model must be invoked. PMID- 15342523 TI - In vivo interaction between mitochondria carrying mtDNAs from different mouse species. AB - Mitochondrial disease model mice, mitomice, were created using zygotes of B6mtspr strain mice carrying mitochondrial DNA (mtDNA) from Mus spretus as recipients of exogenous mitochondria carrying wild-type and a deletion mutant mtDNA (DeltamtDNA) of M. musculus domesticus. In these experiments, mtDNAs from different mouse species were used for identification of exo- and endogenous wild type mtDNAs in the mitomice. Results showed transmission of exogenous DeltamtDNA, but not exogenous wild-type mtDNA, of M. m. domesticus to following generations through the female germ line. Complete elimination of exogenous wild-type mtDNA would be due to stochastic segregation, whereas transmission of exogenous DeltamtDNA would be due to its smaller size leading to a propagational advantage. Tissues in mitomice of the F3 generation carrying exogenous DeltamtDNA showed protection from respiration defects until DeltamtDNA accumulated predominantly. This protection from expression of mitochondrial dysfunction was attained with the help of endogenous wild-type mtDNA of M. spretus, since mitomice did not possess exogenous wild-type mtDNA of M. m. domesticus. These observations provide unambiguous evidence for the presence of interaction between exogenous mitochondria carrying DeltamtDNA and endogenous mitochondria carrying M. spretus wild-type mtDNA. PMID- 15342524 TI - Mapping of multiple quantitative trait loci affecting bovine spongiform encephalopathy. AB - A whole-genome scan was conducted to map quantitative trait loci (QTL) for BSE resistance or susceptibility. Cows from four half-sib families were included and 173 microsatellite markers were used to construct a 2835-cM (Kosambi) linkage map covering 29 autosomes and the pseudoautosomal region of the sex chromosome. Interval mapping by linear regression was applied and extended to a multiple-QTL analysis approach that used identified QTL on other chromosomes as cofactors to increase mapping power. In the multiple-QTL analysis, two genome-wide significant QTL (BTA17 and X/Y(ps)) and four genome-wide suggestive QTL (BTA1, 6, 13, and 19) were revealed. The QTL identified here using linkage analysis do not overlap with regions previously identified using TDT analysis. One factor that may explain the disparity between the results is that a more extensive data set was used in the present study. Furthermore, methodological differences between TDT and linkage analyses may affect the power of these approaches. PMID- 15342525 TI - Evidence for multiple alleles at the DGAT1 locus better explains a quantitative trait locus with major effect on milk fat content in cattle. AB - A quantitative trait locus (QTL) for milk fat percentage has been mapped consistently to the centromeric region of bovine chromosome 14 (BTA14). Two independent studies have identified the nonconservative mutation K232A in the acylCoA-diacylglycerol-acyltransferase 1 (DGAT1) gene as likely to be causal for the observed variation. Here we provide evidence for additional genetic variability at the same QTL that is associated with milk fat percentage variation within the German Holstein population. Namely, we show that alleles of the DGAT1 promoter derived from the variable number of tandem repeat (VNTR) polymorphism are associated with milk fat content in animals homozygous for the allele 232A at DGAT1. Our results present another example for more than two trait-associated alleles being involved in a major gene effect on a quantitative trait. The segregation of multiple alleles affecting milk production traits at the QTL on BTA14 has to be considered whenever marker-assisted selection programs are implemented in dairy cattle. Due to the presence of a potential transcription factor binding site in the 18mer element of the VNTR, the variation in the number of tandem repeats of the 18mer element might be causal for the variability in the transcription level of the DGAT1 gene. PMID- 15342526 TI - Quantitative trait loci associated with adventitious shoot formation in tissue culture and the program of shoot development in Arabidopsis. AB - Arabidopsis ecotypes, Columbia (Col) and Landsberg erecta (Ler), differ in their capacity to regenerate shoots in culture, as do many other cultivars and varieties of the same plant species. Recombinant inbred (RI) lines derived from a cross of Col x Ler were scored for shoot regeneration, and the Arabidopsis genome was scanned using composite interval mapping for loci associated with shoot regeneration. Three QTL were identified--a major one on chromosome 5 in which the Col parent contributed the superior allele and two minor QTL on chromosomes 1 and 4 in which the Ler parent contributed the superior alleles. The RI lines were binned into genotypic pools to isolate the effects of the major QTL on chromosome 5 while holding the minor QTL constant. To identify genes with expression levels that are associated with the allelic state of the major QTL on chromosome 5, oligonucleotide array expression patterns for genes in the LLC pool (Ler alleles at the minor QTL and a Col allele at the major QTL) were compared to those in the LLL pool (Ler alleles at all QTL). The genes that were significantly differentially expressed between the two pools included several encoding transcription factors and signaling or transposon-related proteins. PMID- 15342527 TI - The ULTRAPETALA1 gene functions early in Arabidopsis development to restrict shoot apical meristem activity and acts through WUSCHEL to regulate floral meristem determinacy. AB - Shoot and floral meristem activity in higher plants is controlled by complex signaling networks consisting of positive and negative regulators. The Arabidopsis ULTRAPETALA1 (ULT1) gene has been shown to act as a negative regulator of meristem cell accumulation in inflorescence and floral meristems, as loss-of-function ult1 mutations cause inflorescence meristem enlargement, the production of extra flowers and floral organs, and a decrease in floral meristem determinacy. To investigate whether ULT1 functions in known meristem regulatory pathways, we generated double mutants between ult1 alleles and null alleles of the meristem-promoting genes SHOOTMERISTEMLESS (STM) and WUSCHEL (WUS). We found that, although the ult1 alleles have no detectable embryonic or vegetative phenotypes, ult1 mutations restored extensive organ-forming capability to stm null mutants after germination and increased leaf and floral organ production in stm partial loss-of-function mutants. Mutations in ULT1 also partially suppressed the wus shoot and floral meristem phenotypes. However, wus was epistatic to ult1 in the center of the flower, and WUS transcriptional repression was delayed in ult1 floral meristems. Our results show that during the majority of the Arabidopsis life cycle, ULT1 acts oppositely to STM and WUS in maintaining meristem activity and functions in a separate genetic pathway. However, ULT1 negatively regulates WUS to establish floral meristem determinacy, acting through the WUS-AG temporal feedback loop. PMID- 15342528 TI - Comparative mapping of a major aluminum tolerance gene in sorghum and other species in the poaceae. AB - In several crop species within the Triticeae tribe of the grass family Poaceae, single major aluminum (Al) tolerance genes have been identified that effectively mitigate Al toxicity, a major abiotic constraint to crop production on acidic soils. However, the trait is quantitatively inherited in species within other tribes, and the possible ancestral relationships between major Al tolerance genes and QTL in the grasses remain unresolved. To help establish these relationships, we conducted a molecular genetic analysis of Al tolerance in sorghum and integrated our findings with those from previous studies performed in crop species belonging to different grass tribes. A single locus, AltSB, was found to control Al tolerance in two highly Al tolerant sorghum cultivars. Significant macrosynteny between sorghum and the Triticeae was observed for molecular markers closely linked to putatively orthologous Al tolerance loci present in the group 4 chromosomes of wheat, barley, and rye. However, AltSB was not located within the homeologous region of sorghum but rather mapped near the end of sorghum chromosome 3. Thus, AltSB not only is the first major Al tolerance gene mapped in a grass species that does not belong to the Triticeae, but also appears to be different from the major Al tolerance locus in the Triticeae. Intertribe map comparisons suggest that a major Al tolerance QTL on rice chromosome 1 is likely to be orthologous to AltSB, whereas another rice QTL on chromosome 3 is likely to correspond to the Triticeae group 4 Al tolerance locus. Therefore, this study demonstrates a clear evolutionary link between genes and QTL encoding the same trait in distantly related species within a single plant family. PMID- 15342529 TI - Significance tests and weighted values for AFLP similarities, based on Arabidopsis in silico AFLP fragment length distributions. AB - Many AFLP studies include relatively unrelated genotypes that contribute noise to data sets instead of signal. We developed: (1) estimates of expected AFLP similarities between unrelated genotypes, (2) significance tests for AFLP similarities, enabling the detection of unrelated genotypes, and (3) weighted similarity coefficients, including band position information. Detection of unrelated genotypes and use of weighted similarity coefficients will make the analysis of AFLP data sets more informative and more reliable. Test statistics and weighted coefficients were developed for total numbers of shared bands and for Dice, Jaccard, Nei and Li, and simple matching (dis)similarity coefficients. Theoretical and in silico AFLP fragment length distributions (FLDs) were examined as a basis for the tests. The in silico AFLP FLD based on the Arabidopsis thaliana genome sequence was the most appropriate for angiosperms. The G + C content of the selective nucleotides in the in silico AFLP procedure significantly influenced the FLD. Therefore, separate test statistics were calculated for AFLP procedures with high, average, and low G + C contents in the selective nucleotides. The test statistics are generally applicable for angiosperms with a G + C content of approximately 35-40%, but represent conservative estimates for genotypes with higher G + C contents. For the latter, test statistics based on a rice genome sequence are more appropriate. PMID- 15342530 TI - Transposition of reversed Ac element ends generates chromosome rearrangements in maize. AB - In classical "cut-and-paste" transposition, transposons are excised from donor sites and inserted at new locations. We have identified an alternative pathway in which transposition involves the 5' end of an intact Ac element and the 3' end of a nearby terminally deleted fAc (fractured Ac). The Ac and fAc elements are inserted at the maize p1 locus on chromosome 1s in the same orientation; the adjacent ends of the separate elements are thus in reversed orientation with respect to each other and are separated by a distance of approximately 13 kb. Transposition involving the two ends in reversed orientation generates inversions, deletions, and a novel type of local rearrangement. The rearrangement breakpoints are bounded by the characteristic footprint or target site duplications typical of Ac transposition reactions. These results demonstrate a new intramolecular transposition mechanism by which transposons can greatly impact genome evolution. PMID- 15342531 TI - Allelic and haplotypic diversity at the rp1 rust resistance locus of maize. AB - The maize Rp1 rust resistance locus is a complex consisting of a family of closely related resistance genes. The number of Rp1 paralogs in different maize lines (haplotypes) varied from a single gene in some stocks of the inbred A188 to >50 genes in haplotypes carrying the Rp1-A and Rp1-H specificities. The sequences of paralogs in unrelated haplotypes differ, indicating that the genetic diversity of Rp1-related genes is extremely broad in maize. Two unrelated haplotypes with five or nine paralogs had identical resistance phenotypes (Rp1-D) encoded in genes that differed by three nucleotides resulting in a single amino acid substitution. Genes in some haplotypes are more similar to each other than to any of the genes in other haplotypes indicating that they are evolving in a concerted fashion. PMID- 15342532 TI - The inheritance and evolution of leaf pigmentation and pubescence in teosinte. AB - To investigate the genetic mechanisms that underlie morphological evolution in natural populations, we employed QTL mapping to dissect the inheritance of leaf sheath characters that distinguish Chalco from Balsas teosinte. Abundant macrohairs (trichomes) and intense anthocyanin accumulation are found in Chalco teosinte sheaths whereas Balsas teosinte leaf sheaths are green and glabrous. These character states may represent adaptations to the cooler highland (Chalco) vs. warmer middle-elevation (Balsas) climates. QTL mapping in multiple populations revealed a mix of major- and minor-effect QTL affecting both sheath color (anthocyanin) and macrohair abundance. The major QTL for macrohairs accounts for 52% of the parental difference. Epistatic interactions were detected between the major-effect QTL and multiple other QTL for both traits, accounting for substantial portions of phenotypic variance. Developmental analyses suggest that regulatory program changes underlie the phenotypic differences. Sheath anthocyanin QTL are clearly associated with b1 and a3, both of which are regulators of anthocyanin biosynthesis. Our findings suggest that changes in a small number of QTL can lead to morphological evolution by modulating existing developmental programs. PMID- 15342533 TI - Stochastic and epigenetic changes of gene expression in Arabidopsis polyploids. AB - Polyploidization is an abrupt speciation mechanism for eukaryotes and is especially common in plants. However, little is known about patterns and mechanisms of gene regulation during early stages of polyploid formation. Here we analyzed differential expression patterns of the progenitors' genes among successive selfing generations and independent lineages. The synthetic Arabidopsis allotetraploid lines were produced by a genetic cross between A. thaliana and A. arenosa autotetraploids. We found that some progenitors' genes are differentially expressed in early generations, whereas other genes are silenced in late generations or among different siblings within a selfing generation, suggesting that the silencing of progenitors' genes is rapidly and/or stochastically established. Moreover, a subset of genes is affected in autotetraploid and multiple independent allotetraploid lines and in A. suecica, a natural allotetraploid derived from A. thaliana and A. arenosa, indicating locus specific susceptibility to ploidy-dependent gene regulation. The role of DNA methylation in silencing progenitors' genes is tested in DNA-hypomethylation transgenic lines of A. suecica using RNA interference (RNAi). Two silenced genes are reactivated in both ddm1- and met1-RNAi lines, consistent with the demethylation of centromeric repeats and gene-specific regions in the genome. A rapid and stochastic process of differential gene expression is reinforced by epigenetic regulation during polyploid formation and evolution. PMID- 15342534 TI - Analysis of transposon insertion mutants highlights the diversity of mechanisms underlying male progamic development in Arabidopsis. AB - To identify genes with essential roles in male gametophytic development, including postpollination (progamic) events, we have undertaken a genetic screen based on segregation ratio distortion of a transposon-borne kanamycin-resistance marker. In a population of 3359 Arabidopsis Ds transposon insertion lines, we identified 20 mutants with stably reduced segregation ratios arising from reduced gametophytic transmission. All 20 mutants showed strict cosegregation of Ds and the reduced gametophytic transmission phenotype. Among these, 10 mutants affected both male and female transmission and 10 mutants showed male-specific transmission defects. Four male and female (ungud) mutants and 1 male-specific mutant showed cellular defects in microspores and/or in developing pollen. The 6 remaining ungud mutants and 9 male-specific (seth) mutants affected pollen functions during progamic development. In vitro and in vivo analyses are reported for 5 seth mutants. seth6 completely blocked pollen germination, while seth7 strongly reduced pollen germination efficiency and tube growth. In contrast, seth8, seth9, or seth10 pollen showed reduced competitive ability that was linked to slower rates of pollen tube growth. Gene sequences disrupted in seth insertions suggest essential functions for putative SETH proteins in diverse processes including protein anchoring, cell wall biosynthesis, signaling, and metabolism. PMID- 15342535 TI - Multiple-interval mapping for quantitative trait loci controlling endosperm traits. AB - Endosperm traits are trisomic inheritant and are of great economic importance because they are usually directly related to grain quality. Mapping for quantitative trait loci (QTL) underlying endosperm traits can provide an efficient way to genetically improve grain quality. As the traditional QTL mapping methods (diploid methods) are usually designed for traits under diploid control, they are not the ideal approaches to map endosperm traits because they ignore the triploid nature of endosperm. In this article, a statistical method considering the triploid nature of endosperm (triploid method) is developed on the basis of multiple-interval mapping (MIM) to map for the underlying QTL. The proposed triploid MIM method is derived to broadly use the marker information either from only the maternal plants or from both the maternal plants and their embryos in the backcross and F2 populations for mapping endosperm traits. Due to the use of multiple intervals simultaneously to take multiple QTL into account, the triploid MIM method can provide better detection power and estimation precision, and as shown in this article it is capable of analyzing and searching for epistatic QTL directly as compared to the traditional diploid methods and current triploid methods using only one (or two) interval(s). Several important issues in endosperm trait mapping, such as the relation and differences between the diploid and triploid methods, variance components of genetic variation, and the problems if effects are present and ignored, are also addressed. Simulations are performed to further explore these issues, to investigate the relative efficiency of different experimental designs, and to evaluate the performance of the proposed and current methods in mapping endosperm traits. The MIM-based triploid method can provide a powerful tool to estimate the genetic architecture of endosperm traits and to assist the marker-assisted selection for the improvement of grain quality in crop science. The triploid MIM FORTRAN program for mapping endosperm traits is available on the worldwide web (http://www.stat.sinica.edu.tw/chkao/). PMID- 15342536 TI - Simultaneous detection of linkage disequilibrium and genetic differentiation of subdivided populations. AB - We propose a new method for simultaneously detecting linkage disequilibrium and genetic structure in subdivided populations. Taking subpopulation structure into account with a hierarchical model, we estimate the magnitude of genetic differentiation and linkage disequilibrium in a metapopulation on the basis of geographical samples, rather than decompose a population into a finite number of random-mating subpopulations. We assume that Hardy-Weinberg equilibrium is satisfied in each locality, but do not assume independence between marker loci. Linkage states remain unknown. Genetic differentiation and linkage disequilibrium are expressed as hyperparameters describing the prior distribution of genotypes or haplotypes. We estimate related parameters by maximizing marginal-likelihood functions and detect linkage equilibrium or disequilibrium by the Akaike information criterion. Our empirical Bayesian model analyzes genotype and haplotype frequencies regardless of haploid or diploid data, so it can be applied to most commonly used genetic markers. The performance of our procedure is examined via numerical simulations in comparison with classical procedures. Finally, we analyze isozyme data of ayu, a severely exploited fish species, and single-nucleotide polymorphisms in human ALDH2. PMID- 15342537 TI - Evolution of specialists in an experimental microcosm. AB - The impact of adaptation on the persistence of a balanced polymorphism was explored using the lactose operon of Escherichia coli as a model system. Competition in chemostats for two substitutable resources, methylgalactoside and lactulose, generates stabilizing frequency-dependent selection when two different naturally isolated lac operons (TD2 and TD10) are used. The fate of this balanced polymorphism was tracked over evolutionary time by monitoring the frequency of fhuA-, a linked neutral genetic marker that confers resistance to the bacteriophage T5. In four out of nine chemostats the lac polymorphism persisted for 400-600 generations when the experiments were terminated. In the other five chemostats the fhuA polymorphism, and consequently the lac operon polymorphism, was lost between 86 and 219 generations. Four of 13 chemostat cultures monomorphic for the lac operon retained the neutral fhuA polymorphism for 450-550 generations until they were terminated; the remainder became monomorphic at fhuA between 63 and 303 generations. Specialists on each galactoside were isolated from chemostats that maintained the fhuA polymorphism, whether polymorphic or monomorphic at the lac operon. Strains isolated from three of four chemostats in which the lac polymorphism was preserved had switched their galactoside preference. Most of the chemostats where the fhuA polymorphism was lost also contained specialists. These results demonstrate that the initial polymorphism at lac was of little consequence to the outcome of long-term adaptive evolution. Instead, the fitnesses of evolved strains were dominated by mutations arising elsewhere in the genome, a fact confirmed by showing that operons isolated from their evolved backgrounds were alone unable to explain the presence of both specialists. Our results suggest that, once stabilized, ecological specialization prevented selective sweeps through the entire population, thereby promoting the maintenance of linked neutral polymorphisms. PMID- 15342538 TI - Estimating the frequency of events that cause multiple-nucleotide changes. AB - Existing mathematical models of DNA sequence evolution assume that all substitutions derive from point mutations. There is, however, increasing evidence that larger-scale events, involving two or more consecutive sites, may also be important. We describe a model, denoted SDT, that allows for single-nucleotide, doublet, and triplet mutations. Applied to protein-coding DNA, the SDT model allows doublet and triplet mutations to overlap codon boundaries but still permits data to be analyzed using the simplifying assumption of independence of sites. We have implemented the SDT model for maximum-likelihood phylogenetic inference and have applied it to an alignment of mammalian globin sequences and to 258 other protein-coding sequence alignments from the Pandit database. We find the SDT model's inclusion of doublet and triplet mutations to be overwhelmingly successful in giving statistically significant improvements in fit of model to data, indicating that larger-scale mutation events do occur. Distributions of inferred parameter values over all alignments analyzed suggest that these events are far more prevalent than previously thought. Detailed consideration of our results and the absence of any known mechanism causing three adjacent nucleotides to be substituted simultaneously, however, leads us to suggest that the actual evolutionary events occurring may include still-larger-scale events, such as gene conversion, inversion, or recombination, or a series of rapid compensatory changes. PMID- 15342539 TI - The speed of adaptation in large asexual populations. AB - In large asexual populations, beneficial mutations have to compete with each other for fixation. Here, I derive explicit analytic expressions for the rate of substitution and the mean beneficial effect of fixed mutations, under the assumptions that the population size N is large, that the mean effect of new beneficial mutations is smaller than the mean effect of new deleterious mutations, and that new beneficial mutations are exponentially distributed. As N increases, the rate of substitution approaches a constant, which is equal to the mean effect of new beneficial mutations. The mean effect of fixed mutations continues to grow logarithmically with N. The speed of adaptation, measured as the change of log fitness over time, also grows logarithmically with N for moderately large N, and it grows double-logarithmically for extremely large N. Moreover, I derive a simple formula that determines whether at given N beneficial mutations are expected to compete with each other or go to fixation independently. Finally, I verify all results with numerical simulations. PMID- 15342540 TI - Conditional probability methods for haplotyping in pedigrees. AB - Efficient haplotyping in pedigrees is important for the fine mapping of quantitative trait locus (QTL) or complex disease genes. To reconstruct haplotypes efficiently for a large pedigree with a large number of linked loci, two algorithms based on conditional probabilities and likelihood computations are presented. The first algorithm (the conditional probability method) produces a single, approximately optimal haplotype configuration, with computing time increasing linearly in the number of linked loci and the pedigree size. The other algorithm (the conditional enumeration method) identifies a set of haplotype configurations with high probabilities conditional on the observed genotype data for a pedigree. Its computing time increases less than exponentially with the size of a subset of the set of person-loci with unordered genotypes and linearly with its complement. The size of the subset is controlled by a threshold parameter. The set of identified haplotype configurations can be used to estimate the identity-by-descent (IBD) matrix at a map position for a pedigree. The algorithms have been tested on published and simulated data sets. The new haplotyping methods are much faster and provide more information than several existing stochastic and rule-based methods. The accuracies of the new methods are equivalent to or better than those of these existing methods. PMID- 15342541 TI - Application of coalescent methods to reveal fine-scale rate variation and recombination hotspots. AB - There has been considerable recent interest in understanding the way in which recombination rates vary over small physical distances, and the extent of recombination hotspots, in various genomes. Here we adapt, apply, and assess the power of recently developed coalescent-based approaches to estimating recombination rates from sequence polymorphism data. We apply full-likelihood estimation to study rate variation in and around a well-characterized recombination hotspot in humans, in the beta-globin gene cluster, and show that it provides similar estimates, consistent with those from sperm studies, from two populations deliberately chosen to have different demographic and selectional histories. We also demonstrate how approximate-likelihood methods can be used to detect local recombination hotspots from genomic-scale SNP data. In a simulation study based on 80 100-kb regions, these methods detect 43 out of 60 hotspots (ranging from 1 to 2 kb in size), with only two false positives out of 2000 subregions that were tested for the presence of a hotspot. Our study suggests that new computational tools for sophisticated analysis of population diversity data are valuable for hotspot detection and fine-scale mapping of local recombination rates. PMID- 15342542 TI - Sex-specific meiotic drive and selection at an imprinted locus. AB - We present a one-locus model that breaks two symmetries of Mendelian genetics. Whereas symmetry of transmission is breached by allowing sex-specific segregation distortion, symmetry of expression is breached by allowing genomic imprinting. Simple conditions for the existence of at least one polymorphic stable equilibrium are provided. In general, population mean fitness is not maximized at polymorphic equilibria. However, mean fitness at a polymorphic equilibrium with segregation distortion may be higher than mean fitness at the corresponding equilibrium with Mendelian segregation if one (or both) of the heterozygote classes has higher fitness than both homozygote classes. In this case, mean fitness is maximized by complete, but opposite, drive in the two sexes. We undertook an extensive numerical analysis of the parameter space, finding, for the first time in this class of models, parameter sets yielding two stable polymorphic equilibria. Multiple equilibria exist both with and without genomic imprinting, although they occurred in a greater proportion of parameter sets with genomic imprinting. PMID- 15342543 TI - Maximum-likelihood estimation of rates of recombination within mating-type regions. AB - Features common to many mating-type regions include recombination suppression over large genomic tracts and cosegregation of genes of various functions, not necessarily related to reproduction. Model systems for homomorphic self incompatibility (SI) in flowering plants share these characteristics. We introduce a method for the exact computation of the joint probability of numbers of neutral mutations segregating at the determinant of mating type and at a linked marker locus. The underlying Markov model incorporates strong balancing selection into a two-locus coalescent. We apply the method to obtain a maximum likelihood estimate of the rate of recombination between a marker locus, 48A, and S-RNase, the determinant of SI specificity in pistils of Nicotiana alata. Even though the sampled haplotypes show complete allelic linkage disequilibrium and recombinants have never been detected, a highly significant deficiency of synonymous substitutions at 48A compared to S-RNase suggests a history of recombination. Our maximum-likelihood estimate indicates a rate of recombination of perhaps 3 orders of magnitude greater than the rate of synonymous mutation. This approach may facilitate the construction of genetic maps of regions tightly linked to targets of strong balancing selection. PMID- 15342544 TI - Genome image programs: visualization and interpretation of Escherichia coli microarray experiments. AB - We have developed programs to facilitate analysis of microarray data in Escherichia coli. They fall into two categories: manipulation of microarray images and identification of known biological relationships among lists of genes. A program in the first category arranges spots from glass-slide DNA microarrays according to their position in the E. coli genome and displays them compactly in genome order. The resulting genome image is presented in a web browser with an image map that allows the user to identify genes in the reordered image. Another program in the first category aligns genome images from two or more experiments. These images assist in visualizing regions of the genome with common transcriptional control. Such regions include multigene operons and clusters of operons, which are easily identified as strings of adjacent, similarly colored spots. The images are also useful for assessing the overall quality of experiments. The second category of programs includes a database and a number of tools for displaying biological information about many E. coli genes simultaneously rather than one gene at a time, which facilitates identifying relationships among them. These programs have accelerated and enhanced our interpretation of results from E. coli DNA microarray experiments. Examples are given. PMID- 15342545 TI - Genes encoding subunits of stable complexes are clustered on the yeast chromosomes: an interpretation from a dosage balance perspective. AB - Genomic evidence for colocalization of functionally related genes on eukaryote chromosomes is mounting. Here we show that a statistically significant fraction of yeast genes coding for subunits of stable complexes are located within 10-30 kb of each other. Clustering of genes encoding subunits of complexes may ensure better coregulation and maintain the right stoichiometry of complexes upon duplication of chromosomal segments. PMID- 15342546 TI - A potential regulatory polymorphism upstream of hairy is not associated with bristle number variation in wild-caught Drosophila. AB - To extend results from laboratory genetic mapping experiments to natural populations it is necessary to estimate the phenotypic effects attributable to laboratory-identified genetic factors in nature. We retested a polymorphism found to be strongly associated with an increase of 0.35 sternopleural bristles in laboratory strains in two large samples of wild-caught Drosophila melanogaster. Despite >90% power to detect effects as low as 0.27 bristles (<1% of the total variation in bristle number) we did not replicate the association in nature. Potential explanations for this result are explored. PMID- 15342547 TI - A fast algorithm for functional mapping of complex traits. AB - By integrating the underlying developmental mechanisms for the phenotypic formation of traits into a mapping framework, functional mapping has emerged as an important statistical approach for mapping complex traits. In this note, we explore the feasibility of using the simplex algorithm as an alternative to solve the mixture-based likelihood for functional mapping of complex traits. The results from the simplex algorithm are consistent with those from the traditional EM algorithm, but the simplex algorithm has considerably reduced computational times. Moreover, because of its nonderivative nature and easy implementation with current software, the simplex algorithm enjoys an advantage over the EM algorithm in the dynamic modeling and analysis of complex traits. PMID- 15342548 TI - Commentary on Klar. PMID- 15342550 TI - Glycoprofiling with micro-arrays of glycoconjugates and lectins. AB - To facilitate deciphering the information content in the glycome, thin film coated photoactivatable surfaces were applied for covalent immobilization of glycans, glycoconjugates, or lectins in microarray formats. Light-induced immobilization of a series of bacterial exopolysaccharides on photoactivatable dextran-coated analytical platforms allowed covalent binding of the exopolysaccharides. Their specific galactose decoration was detected with fluorescence-labeled lectins. Similarly, glycoconjugates were covalently immobilized and displayed glycans were profiled for fucose, sialic acid, galactose, and lactosamine epitopes. The applicability of such platforms for glycan profiling was further tested with extracts of Caco2 epithelial cells. Following spontaneous differentiation or on pretreatment with sialyllactose, Caco2 cells showed a reduction of specific glycan epitopes. The changed glycosylation phenotypes coincided with altered enteropathogenic E. coli adhesion to the cells. This microarray strategy was also suitable for the immobilization of lectins through biotin-neutravidin-biotin bridging on platforms functionalized with a biotin derivatized photoactivatable dextran. All immobilized glycans were specifically and differentially detected either on glycoconjugate or lectin arrays. The results demonstrate the feasibility and versatility of the novel platforms for glycan profiling. PMID- 15342551 TI - Massive accumulation of Man2GlcNAc2-Asn in nonneuronal tissues of glycosylasparaginase-deficient mice and its removal by enzyme replacement therapy. AB - Aspartylglycosaminuria (AGU) is caused by deficient enzymatic activity of glycosylasparaginase (GA). The disease is characterized by accumulation of aspartylglucosamine (GlcNAc-Asn) and other glycoasparagines in tissues and body fluids of AGU patients and in an AGU mouse model. In the current study, we characterized a glycoasparagine carrying the tetrasaccharide moiety of alpha-D Man-(1-->6)-beta-D-Man-(1-->4)-beta-D-GlcNAc-(1-->4)-beta-D-GlcNAc-(1-->N)-Asn (Man2GlcNAc2-Asn) in urine of an AGU patient and also in the tissues of the AGU mouse model. Quantitative analysis demonstrated a massive accumulation of the compound especially in nonneuronal tissues of the AGU mice, in which the levels of Man2GlcNAc2-Asn were typically 30-87% of those of GlcNAc-Asn. The highest level of Man2GlcNAc2-Asn was found in the liver, spleen, and heart tissues of the AGU mice, the respective amounts being 87%, 76%, and 57% of the GlcNAc-Asn levels. In the brain tissue of AGU mice the Man2GlcNAc2-Asn storage was only 9% of that of GlcNAc-Asn. In contrast to GlcNAc-Asn, the storage of Man2GlcNAc2-Asn markedly increased in the liver and spleen tissues of AGU mice as they grew older. Enzyme replacement therapy with glycosylasparaginase for 3.5 weeks reduced the amount of Man2GlcNAc2-Asn by 66-97% in nonneuronal tissues, but only by 13% in the brain tissue of the AGU mice. In conclusion, there is evidence for a role for storage of glycoasparagines other than aspartylglucosamine in the pathogenesis of AGU, and this possibility should be taken into consideration in the treatment of the disease. PMID- 15342552 TI - Anti-alpha-galactosyl antibodies recognizing epitopes terminating with alpha1,4 linked galactose: human natural and mouse monoclonal anti-NOR and anti-P1 antibodies. AB - The rare NOR erythrocytes, which are agglutinated by most human sera, contain unique glycosphingolipids (globoside elongation products) terminating with the sequence Galalpha1-4GalNAcbeta1-3Gal- recognized by common natural human antibodies. Anti-NOR antibodies were isolated from several human sera by affinity procedures, and their specificity was tested by inhibition of antibody binding to NOR-tri-polyacrylamide (PAA) conjugate (ELISA) by the synthetic oligosaccharides, Galalpha1-4GalNAcbeta1-3Gal (NOR-tri), Galalpha1-4GalNAc (NOR-di), Galalpha1 4Galbeta1-3Galbeta1-4Glc ((Gal)3Glc), and Galalpha1-4Gal (P1-di). Two major types of subspecificity of anti-NOR antibodies were found. Type 1 antibodies were found to react strongly with (Gal)3Glc and NOR-tri and weakly with P1-di and NOR-di, which indicated specificity for the trisaccharide epitope Galalpha1 4Gal/GalNAcbeta1-3Gal. Type 2 antibodies were specific to Galalpha1-4GalNAc, because they were inhibited most strongly by NOR-tri and NOR-di and were not (or very weakly) inhibited by (Gal)3Glc and P1-di. Monoclonal anti-NOR antibodies were obtained by immunizing mice with NOR-tri-human serum albumin (HSA) conjugate and were found to have type 2 specificity. All anti-NOR antibodies reacted specifically with NOR glycolipids on thin-layer plates. The cross-reactivity of type 1 anti-NOR antibodies with Galalpha1-4Gal drew attention to a possible antigenic relationship between NOR and blood group P system glycolipids. The latter glycolipids include Pk (Galalpha1-4Galbeta1-4Glc-Cer) present in all normal erythrocytes and P1 (Galalpha1-4Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc-Cer) present only in P1 erythrocytes. Sera of some P2 (P1-negative) persons contain natural anti-P1 antibodies. This prompted us to test the specificity of anti-P1 antibodies. Natural human anti-P1 isolated from serum of P2 individual and mouse monoclonal anti-P1 were best inhibited by Galalpha1-4Galbeta1-4GlcNAc (P1-tri) and did not react with NOR-tri and NOR-di. Monoclonal anti-P1 bound to Pk and P1 glycolipids and not to NOR glycolipids. These results indicated an entirely different specificity of anti-NOR and anti-P1 antibodies. Human serum samples differed in the content of anti-alpha-galactosyl antibodies, including both types of anti-NOR. In the sera of some individuals, type 1 or type 2 anti-NOR antibodies dominated, and other samples contained mixtures of both types of anti NOR. The biological significance of these new abundant anti-alpha-galactosyl antibodies still awaits elucidation. PMID- 15342553 TI - Evidence for gradients of human genetic diversity within and among continents. AB - Genetic variation in humans is sometimes described as being discontinuous among continents or among groups of individuals, and by some this has been interpreted as genetic support for "races." A recent study in which >350 microsatellites were studied in a global sample of humans showed that they could be grouped according to their continental origin, and this was widely interpreted as evidence for a discrete distribution of human genetic diversity. Here, we investigate how study design can influence such conclusions. Our results show that when individuals are sampled homogeneously from around the globe, the pattern seen is one of gradients of allele frequencies that extend over the entire world, rather than discrete clusters. Therefore, there is no reason to assume that major genetic discontinuities exist between different continents or "races." PMID- 15342554 TI - Comparative analysis of apicomplexa and genomic diversity in eukaryotes. AB - The apicomplexans Plasmodium and Cryptosporidium have developed distinctive adaptations via lineage-specific gene loss and gene innovation in the process of diverging from a common parasitic ancestor. The two lineages have acquired distinct but overlapping sets of surface protein adhesion domains typical of animal proteins, but in no case do they share multidomain architectures identical to animals. Cryptosporidium, but not Plasmodium, possesses an animal-type O linked glycosylation pathway, along with >30 predicted surface proteins having mucin-like segments. The two parasites have notable qualitative differences in conserved protein architectures associated with chromatin dynamics and transcription. Cryptosporidium shows considerable reduction in the number of introns and a concomitant loss of spliceosomal machinery components. We also describe additional molecular characteristics distinguishing Apicomplexa from other eukaryotes for which complete genome sequences are available. PMID- 15342555 TI - Recurrent sites for new centromere seeding. AB - Using comparative FISH and genomics, we have studied and compared the evolution of chromosome 3 in primates and two human neocentromere cases on the long arm of this chromosome. Our results show that one of the human neocentromere cases maps to the same 3q26 chromosomal region where a new centromere emerged in a common ancestor of the Old World monkeys approximately 25-40 million years ago. Similarly, the locus in which a new centromere was seeded in the great apes' ancestor was orthologous to the site in which a new centromere emerged in the New World monkeys' ancestor. These data suggest the recurrent use of longstanding latent centromeres and that there is an inherent potential of these regions to form centromeres. The second human neocentromere case (3q24) revealed unprecedented features. The neocentromere emergence was not accompanied by any chromosomal rearrangement that usually triggers these events. Instead, it involved the functional inactivation of the normal centromere, and was present in an otherwise phenotypically normal individual who transmitted this unusual chromosome to the next generation. We propose that the formation of neocentromeres in humans and the emergence of new centromeres during the course of evolution share a common mechanism. PMID- 15342557 TI - Widespread RNA editing of embedded alu elements in the human transcriptome. AB - More than one million copies of the approximately 300-bp Alu element are interspersed throughout the human genome, with up to 75% of all known genes having Alu insertions within their introns and/or UTRs. Transcribed Alu sequences can alter splicing patterns by generating new exons, but other impacts of intragenic Alu elements on their host RNA are largely unexplored. Recently, repeat elements present in the introns or 3'-UTRs of 15 human brain RNAs have been shown to be targets for multiple adenosine to inosine (A-to-I) editing. Using a statistical approach, we find that editing of transcripts with embedded Alu sequences is a global phenomenon in the human transcriptome, observed in 2674 ( approximately 2%) of all publicly available full-length human cDNAs (n = 128,406), from >250 libraries and >30 tissue sources. In the vast majority of edited RNAs, A-to-I substitutions are clustered within transcribed sense or antisense Alu sequences. Edited bases are primarily associated with retained introns, extended UTRs, or with transcripts that have no corresponding known gene. Therefore, Alu-associated RNA editing may be a mechanism for marking nonstandard transcripts, not destined for translation. PMID- 15342559 TI - EGPred: prediction of eukaryotic genes using ab initio methods after combining with sequence similarity approaches. AB - EGPred is a Web-based server that combines ab initio methods and similarity searches to predict genes, particularly exon regions, with high accuracy. The EGPred program proceeds in the following steps: (1) an initial BLASTX search of genomic sequence against the RefSeq database is used to identify protein hits with an E-value <1; (2) a second BLASTX search of genomic sequence against the hits from the previous run with relaxed parameters (E-values <10) helps to retrieve all probable coding exon regions; (3) a BLASTN search of genomic sequence against the intron database is then used to detect probable intron regions; (4) the probable intron and exon regions are compared to filter/remove wrong exons; (5) the NNSPLICE program is then used to reassign splicing signal site positions in the remaining probable coding exons; and (6) finally ab initio predictions are combined with exons derived from the fifth step based on the relative strength of start/stop and splice signal sites as obtained from ab initio and similarity search. The combination method increases the exon level performance of five different ab initio programs by 4%-10% when evaluated on the HMR195 data set. Similar improvement is observed when ab initio programs are evaluated on the Burset/Guigo data set. Finally, EGPred is demonstrated on an approximately 95-Mbp fragment of human chromosome 13. The list of predicted genes from this analysis are available in the supplementary material. The EGPred program is computationally intensive due to multiple BLAST runs during each analysis. The EGPred server is available at http://www.imtech.res.in/raghava/egpred/. PMID- 15342558 TI - Phylogenetic footprint analysis of IGF2 in extant mammals. AB - Genomic imprinting results in monoallelic gene transcription that is directed by cis-acting regulatory elements epigenetically marked in a parent-of-origin dependent manner. We performed phylogenetic sequence and epigenetic comparisons of IGF2 between the nonimprinted platypus (Ornithorhynchus anatinus) and imprinted opossum (Didelphis virginiana), mouse (Mus musculus), and human (Homo sapiens) to determine if their divergent imprint status would reflect differences in the conservation of genomic elements important in the regulation of imprinting. We report herein that IGF2 imprinting does not correlate evolutionarily with differential intragenic methylation, nor is it associated with motif 13, a reported IGF2-specific "imprint signature" located in the coding region. Instead, IGF2 imprinting is strongly associated with both the lack of short interspersed transposable elements (SINEs) and an intragenic conserved inverted repeat that contains candidate CTCF-binding sites, a role not previously ascribed to this particular sequence element. Our results are the first to demonstrate that comparative footprint analysis of species from evolutionarily distant mammalian clades, and exhibiting divergent imprint status is a powerful bioinformatics-based approach for identifying cis-acting elements potentially involved not only in the origins of genomic imprinting, but also in its maintenance in humans. PMID- 15342556 TI - Sequence comparison of human and mouse genes reveals a homologous block structure in the promoter regions. AB - Comparative sequence analysis was carried out for the regions adjacent to experimentally validated transcriptional start sites (TSSs), using 3324 pairs of human and mouse genes. We aligned the upstream putative promoter sequences over the 1-kb proximal regions and found that the sequence conservation could not be further extended at, on average, 510 bp upstream positions of the TSSs. This discontinuous manner of the sequence conservation revealed a "block" structure in about one-third of the putative promoter regions. Consistently, we also observed that G+C content and CpG frequency were significantly different inside and outside the blocks. Within the blocks, the sequence identity was uniformly 65% regardless of their length. About 90% of the previously characterized transcription factor binding sites were located within those blocks. In 46% of the blocks, the 5' ends were bounded by interspersed repetitive elements, some of which may have nucleated the genomic rearrangements. The length of the blocks was shortest in the promoters of genes encoding transcription factors and of genes whose expression patterns are brain specific, which suggests that the evolutional diversifications in the transcriptional modulations should be the most marked in these populations of genes. PMID- 15342560 TI - A benchmark for methods in reverse engineering and model discrimination: problem formulation and solutions. AB - A benchmark problem is described for the reconstruction and analysis of biochemical networks given sampled experimental data. The growth of the organisms is described in a bioreactor in which one substrate is fed into the reactor with a given feed rate and feed concentration. Measurements for some intracellular components are provided representing a small biochemical network. Problems of reverse engineering, parameter estimation, and identifiability are addressed. The contribution mainly focuses on the problem of model discrimination. If two or more model variants describe the available experimental data, a new experiment must be designed to discriminate between the hypothetical models. For the problem presented, the feed rate and feed concentration of a bioreactor system are available as control inputs. To verify calculated input profiles an interactive Web site (http://www.sysbio.de/projects/benchmark/) is provided. Several solutions based on linear and nonlinear models are discussed. PMID- 15342561 TI - De novo repeat classification and fragment assembly. AB - Repetitive sequences make up a significant fraction of almost any genome, and an important and still open question in bioinformatics is how to represent all repeats in DNA sequences. We propose a new approach to repeat classification that represents all repeats in a genome as a mosaic of sub-repeats. Our key algorithmic idea also leads to new approaches to multiple alignment and fragment assembly. In particular, we show that our FragmentGluer assembler improves on Phrap and ARACHNE in assembly of BACs and bacterial genomes. PMID- 15342562 TI - Genome-scale in silico models of E. coli have multiple equivalent phenotypic states: assessment of correlated reaction subsets that comprise network states. AB - The constraint-based analysis of genome-scale metabolic and regulatory networks has been successful in predicting phenotypes and useful for analyzing high throughput data sets. Within this modeling framework, linear optimization has been used to study genome-scale metabolic models, resulting in the enumeration of single optimal solutions describing the best use of the network to support growth. Here mixed-integer linear programming was used to calculate and study a subset of the alternate optimal solutions for a genome-scale metabolic model of Escherichia coli (iJR904) under a wide variety of environmental conditions. Analysis of the calculated sets of optimal solutions found that: (1) only a small subset of reactions in the network have variable fluxes across optima; (2) sets of reactions that are always used together in optimal solutions, correlated reaction sets, showed moderate agreement with the currently known transcriptional regulatory structure in E. coli and available expression data, and (3) reactions that are used under certain environmental conditions can provide clues about network regulatory needs. In addition, calculation of suboptimal flux distributions, using flux variability analysis, identified reactions which are used under significantly more environmental conditions suboptimally than optimally. Together these results demonstrate the utilization of reactions in genome-scale models under a variety of different growth conditions. PMID- 15342563 TI - An efficient SNP system for mouse genome scanning and elucidating strain relationships. AB - A set of 1638 informative SNP markers easily assayed by the Amplifluor genotyping system were tested in 102 mouse strains, including the majority of the common and wild-derived inbred strains available from The Jackson Laboratory. Selected from publicly available databases, the markers are on average approximately 1.5 Mb apart and, whenever possible, represent the rare allele in at least two strains. Amplifluor assays were developed for each marker and performed on two independent DNA samples from each strain. The mean number of polymorphisms between strains was 608+/-136 SD. Several tests indicate that the markers provide an effective system for performing genome scans and quantitative trait loci analyses in all but the most closely related strains. Additionally, the markers revealed several subtle differences between closely related mouse strains, including the groups of several 129, BALB, C3H, C57, and DBA strains, and a group of wild-derived inbred strains representing several Mus musculus subspecies. Applying a neighbor-joining method to the data, we constructed a mouse strain family tree, which in most cases confirmed existing genealogies. PMID- 15342564 TI - An SNP resource for rice genetics and breeding based on subspecies indica and japonica genome alignments. AB - Dense coverage of the rice genome with polymorphic DNA markers is an invaluable tool for DNA marker-assisted breeding, positional cloning, and a wide range of evolutionary studies. We have aligned drafts of two rice subspecies, indica and japonica, and analyzed levels and patterns of genetic diversity. After filtering multiple copy and low quality sequence, 408,898 candidate DNA polymorphisms (SNPs/INDELs) were discerned between the two subspecies. These filters have the consequence that our data set includes only a subset of the available SNPs (in particular excluding large numbers of SNPs that may occur between repetitive DNA alleles) but increase the likelihood that this subset is useful: Direct sequencing suggests that 79.8% +/- 7.5% of the in silico SNPs are real. The SNP sample in our database is not randomly distributed across the genome. In fact, 566 rice genomic regions had unusually high (328 contigs/48.6 Mb/13.6% of genome) or low (237 contigs/64.7 Mb/18.1% of genome) polymorphism rates. Many SNP-poor regions were substantially longer than most SNP-rich regions, covering up to 4 Mb, and possibly reflecting introgression between the respective gene pools that may have occurred hundreds of years ago. Although 46.2% +/- 8.3% of the SNPs differentiate other pairs of japonica and indica genotypes, SNP rates in rice were not predictive of evolutionary rates for corresponding genes in another grass species, sorghum. The data set is freely available at http://www.plantgenome.uga.edu/snp. PMID- 15342565 TI - DNA toroids: framework for DNA repair in Deinococcus radiodurans and in germinating bacterial spores. PMID- 15342566 TI - The architecture of the murein (peptidoglycan) in gram-negative bacteria: vertical scaffold or horizontal layer(s)? PMID- 15342567 TI - The Sinorhizobium meliloti ABC transporter Cho is highly specific for choline and expressed in bacteroids from Medicago sativa nodules. AB - In Sinorhizobium meliloti, choline is the direct precursor of phosphatidylcholine, a major lipid membrane component in the Rhizobiaceae family, and glycine betaine, an important osmoprotectant. Moreover, choline is an efficient energy source which supports growth. Using a PCR strategy, we identified three chromosomal genes (choXWV) which encode components of an ABC transporter: ChoX (binding protein), ChoW (permease), and ChoV (ATPase). Whereas the best homology scores were obtained with components of betaine ProU-like systems, Cho is not involved in betaine transport. Site-directed mutagenesis of choX strongly reduced (60 to 75%) the choline uptake activity, and purification of ChoX, together with analysis of the ligand-binding specificity, showed that ChoX binds choline with a high affinity (KD, 2.7 microM) and acetylcholine with a low affinity (KD, 145 microM) but binds none of the betaines. Uptake competition experiments also revealed that ectoine, various betaines, and choline derivatives were not effective competitors for Cho-mediated choline transport. Thus, Cho is a highly specific high-affinity choline transporter. Choline transport activity and ChoX expression were induced by choline but not by salt stress. Western blotting experiments with antibodies raised against ChoX demonstrated the presence of ChoX in bacteroids isolated from nitrogen-fixing nodules obtained from Medicago sativa roots. The choX mutation did not have an effect on growth under standard conditions, and neither Nod nor Fix phenotypes were impaired in the mutant, suggesting that the remaining choline uptake system(s) still present in the mutant strain can compensate for the lack of Cho transporter. PMID- 15342568 TI - Regulation of the intracellular free iron pool by Dpr provides oxygen tolerance to Streptococcus mutans. AB - Dpr is an iron-binding protein required for oxygen tolerance in Streptococcus mutans. We previously proposed that Dpr could confer oxygen tolerance to the bacterium by sequestering intracellular free iron ions that catalyze generation of highly toxic radicals (Y. Yamamoto, M. Higuchi, L. B. Poole, and Y. Kamio, J. Bacteriol. 182:3740-3747, 2000; Y. Yamamoto, L. B. Poole, R. R. Hantgan, and Y. Kamio, J. Bacteriol. 184:2931-2939, 2002). Here, we examined the intracellular free iron status of wild-type (WT) and dpr mutant strains of S. mutans, before and after exposure to air, by using electron spin resonance spectrometry. Under anaerobic conditions, free iron ion concentrations of WT and dpr strains were 225.9 +/- 2.6 and 333.0 +/- 61.3 microM, respectively. Exposure of WT cells to air for 1 h induced Dpr expression and reduced intracellular free iron ion concentrations to 22.5 +/- 5.3 microM; under these conditions, dpr mutant cells maintained intracellular iron concentration at 230.3 +/- 28.8 microM. A decrease in cell viability and genomic DNA degradation was observed in the dpr mutant exposed to air. These data indicate that regulation of the intracellular free iron pool by Dpr is required for oxygen tolerance in S. mutans. PMID- 15342569 TI - DegU-P represses expression of the motility fla-che operon in Bacillus subtilis. AB - Bacillus subtilis implements several adaptive strategies to cope with nutrient limitation experienced at the end of exponential growth. The DegS-DegU two component system is part of the network involved in the regulation of postexponential responses, such as competence development, the production of exoenzymes, and motility. The degU32(Hy) mutation extends the half-life of the phosphorylated form of DegU (DegU-P); this in turn increases the production of alkaline protease, levan-sucrase, and other exoenzymes and inhibits motility and the production of flagella. The expression of the flagellum-specific sigma factor SigD, of the flagellin gene hag, and of the fla-che operon is strongly reduced in a degU32(Hy) genetic background. To investigate the mechanism of action of DegU-P on motility, we isolated mutants of degU32(Hy) that completely suppressed the motility deficiency. The mutations were genetically mapped and characterized by PCR and sequencing. Most of the mutations were found to delete a transcriptional termination signal upstream of the main flagellar operon, fla-che, thus allowing transcriptional readthrough from the cod operon. Two additional mutations improved the sigmaA-dependent promoter sequence of the fla-che operon. Using an electrophoretic mobility shift assay, we have demonstrated that purified DegU binds specifically to the PA promoter region of the fla-che operon. The data suggest that DegU represses transcription of the fla-che operon, and they indicate a central role of the operon in regulating the synthesis and assembly of flagella. PMID- 15342570 TI - Involvement of a plasmid-encoded type IV secretion system in the plant tissue watersoaking phenotype of Burkholderia cenocepacia. AB - Burkholderia cenocepacia strain K56-2, a representative of the Burkholderia cepacia complex, is part of the epidemic and clinically problematic ET12 lineage. The strain produced plant tissue watersoaking (ptw) on onion tissue, which is a plant disease-associated trait. Using plasposon mutagenesis, mutants in the ptw phenotype were generated. The translated sequence of a disrupted gene (ptwD4) from a ptw-negative mutant showed homology to VirD4-like proteins. Analysis of the region proximal to the transfer gene homolog identified a gene cluster located on the 92-kb resident plasmid that showed homology to type IV secretion systems. The role of ptwD4, ptwC, ptwB4, and ptwB10 in the expression of ptw activity was determined by conducting site-directed mutagenesis. The ptw phenotype was not expressed by K56-2 derivatives with a disruption in ptwD4, ptwB4, or ptwB10 but was observed in a derivative with a disruption in ptwC. Complementation of ptw-negative K56-2 derivatives in trans resulted in complete restoration of the ptw phenotype. In addition, analysis of culture supernatants revealed that the putative ptw effector(s) was a secreted, heat-stable protein(s) that caused plasmolysis of plant protoplasts. A second chromosomally encoded type IV secretion system with complete homology to the VirB-VirD system was identified in K56-2. Site-directed mutagenesis of key secretory genes in the VirB-VirD system did not affect expression of the ptw phenotype. Our findings indicate that in strain K56-2, the plasmid-encoded Ptw type IV secretion system is responsible for the secretion of a plant cytotoxic protein(s). PMID- 15342571 TI - Denitrification genes regulate Brucella virulence in mice. AB - Brucella is the causative agent of the zoonotic disease brucellosis, which is endemic in many parts of the world. Genome sequencing of B. suis and B. melitensis revealed that both are complete denitrifiers. To learn more about the role of denitrification in these animal pathogens, a study of the role of denitrification in the closely related B. neotomae was undertaken. In contrast to B. suis and B. melitensis, it was found that B. neotomae is a partial denitrifier that can reduce nitrate to nitrite but no further. Examination of the B. neotomae genome showed that a deletion in the denitrification gene cluster resulted in complete loss of nirV and the partial deletion of nirK and nnrA. Even though the nor operon is intact, a norC-lacZ promoter fusion was not expressed in B. neotomae. However, the norC-lacZ fusion was expressed in the related denitrifier Agrobacterium tumefaciens, suggesting that the lack of expression in B. neotomae is due to inactivation of NnrA. A narK-lacZ promoter fusion was found to exhibit nitrate-dependent expression consistent with the partial denitrifier phenotype. Complementation of the deleted region in B. neotomae by using nirK, nirV, and nnrA from B. melitensis restored the ability of B. neotomae to reduce nitrite. There was a significant difference in the death of IRF-1-/- mice when infected with B. neotomae containing nirK, nirV, and nnrA and those infected with wild type B. neotomae. The wild-type strain killed all the infected mice, whereas most of the mice infected with B. neotomae containing nirK, nirV, and nnrA survived. PMID- 15342572 TI - Escherichia coli glutamate- and arginine-dependent acid resistance systems increase internal pH and reverse transmembrane potential. AB - Due to the acidic nature of the stomach, enteric organisms must withstand extreme acid stress for colonization and pathogenesis. Escherichia coli contains several acid resistance systems that protect cells to pH 2. One acid resistance system, acid resistance system 2 (AR2), requires extracellular glutamate, while another (AR3) requires extracellular arginine. Little is known about how these systems protect cells from acid stress. AR2 and AR3 are thought to consume intracellular protons through amino acid decarboxylation. Antiport mechanisms then exchange decarboxylation products for new amino acid substrates. This form of proton consumption could maintain an internal pH (pHi) conducive to cell survival. The model was tested by estimating the pHi and transmembrane potential (DeltaPsi) of cells acid stressed at pH 2.5. During acid challenge, glutamate- and arginine dependent systems elevated pHi from 3.6 to 4.2 and 4.7, respectively. However, when pHi was manipulated to 4.0 in the presence or absence of glutamate, only cultures challenged in the presence of glutamate survived, indicating that a physiological parameter aside from pHi was also important. Measurements of DeltaPsi indicated that amino acid-dependent acid resistance systems help convert membrane potential from an inside negative to inside positive charge, an established acidophile strategy used to survive extreme acidic environments. Thus, reversing DeltaPsi may be a more important acid resistance strategy than maintaining a specific pHi value. PMID- 15342573 TI - Sinorhizobium meliloti ExoR and ExoS proteins regulate both succinoglycan and flagellum production. AB - The production of the Sinorhizobium meliloti exopolysaccharide, succinoglycan, is required for the formation of infection threads inside root hairs, a critical step during the nodulation of alfalfa (Medicago sativa) by S. meliloti. Two bacterial mutations, exoR95::Tn5 and exoS96::Tn5, resulted in the overproduction of succinoglycan and a reduction in symbiosis. Systematic analyses of the symbiotic phenotypes of the two mutants demonstrated their reduced efficiency of root hair colonization. In addition, both the exoR95 and exoS96 mutations caused a marked reduction in the biosynthesis of flagella and consequent loss of ability of the cells to swarm and swim. Succinoglycan overproduction did not appear to be the cause of the suppression of flagellum biosynthesis. Further analysis indicated that both the exoR95 and exoS96 mutations affected the expression of the flagellum biosynthesis genes. These findings suggest that both the ExoR protein and the ExoS/ChvI two-component regulatory system are involved in the regulation of both succinoglycan and flagellum biosynthesis. These findings provide new avenues of understanding of the physiological changes S. meliloti cells go through during the early stages of symbiosis and of the signal transduction pathways that mediate such changes. PMID- 15342574 TI - Targeted mutagenesis of the Mycobacterium smegmatis mca gene, encoding a mycothiol-dependent detoxification protein. AB - Mycothiol (MSH), a functional analogue of glutathione (GSH) that is found exclusively in actinomycetes, reacts with electrophiles and toxins to form MSH toxin conjugates. Mycothiol S-conjugate amidase (Mca) then catalyzes the hydrolysis of an amide bond in the S conjugates, producing a mercapturic acid of the toxin, which is excreted from the bacterium, and glucosaminyl inositol, which is recycled back to MSH. In this study, we have generated and characterized an allelic exchange mutant of the mca gene of Mycobacterium smegmatis. The mca mutant accumulates the S conjugates of the thiol-specific alkylating agent monobromobimane and the antibiotic rifamycin S. Introduction of M. tuberculosis mca epichromosomally or introduction of M. smegmatis mca integratively resulted in complementation of Mca activity and reduced levels of S conjugates. The mutation in mca renders the mutant strain more susceptible to electrophilic toxins, such as N-ethylmalemide, iodoacetamide, and chlorodinitrobenzene, and to several oxidants, such as menadione and plumbagin. Additionally we have shown that the mca mutant is also more susceptible to the antituberculous antibiotic streptomycin. Mutants disrupted in genes belonging to MSH biosynthesis are also more susceptible to streptomycin, providing further evidence that Mca detoxifies streptomycin in the mycobacterial cell in an MSH-dependent manner. PMID- 15342575 TI - Two arginine repressors regulate arginine biosynthesis in Lactobacillus plantarum. AB - The repression of the carAB operon encoding carbamoyl phosphate synthase leads to Lactobacillus plantarum FB331 growth inhibition in the presence of arginine. This phenotype was used in a positive screening to select spontaneous mutants deregulated in the arginine biosynthesis pathway. Fourteen mutants were genetically characterized for constitutive arginine production. Mutations were located either in one of the arginine repressor genes (argR1 or argR2) present in L. plantarum or in a putative ARG operator in the intergenic region of the bipolar carAB-argCJBDF operons involved in arginine biosynthesis. Although the presence of two ArgR regulators is commonly found in gram-positive bacteria, only single arginine repressors have so far been well studied in Escherichia coli or Bacillus subtilis. In L. plantarum, arginine repression was abolished when ArgR1 or ArgR2 was mutated in the DNA binding domain, or in the oligomerization domain or when an A123D mutation occurred in ArgR1. A123, equivalent to the conserved residue A124 in E. coli ArgR involved in arginine binding, was different in the wild-type ArgR2. Thus, corepressor binding sites may be different in ArgR1 and ArgR2, which have only 35% identical residues. Other mutants harbored wild-type argR genes, and 20 mutants have lost their ability to grow in normal air without carbon dioxide enrichment; this revealed a link between arginine biosynthesis and a still-unknown CO2-dependent metabolic pathway. In many gram-positive bacteria, the expression and interaction of different ArgR-like proteins may imply a complex regulatory network in response to environmental stimuli. PMID- 15342576 TI - Among multiple phosphomannomutase gene orthologues, only one gene encodes a protein with phosphoglucomutase and phosphomannomutase activities in Thermococcus kodakaraensis. AB - Four orthologous genes (TK1108, TK1404, TK1777, and TK2185) that can be annotated as phosphomannomutase (PMM) genes (COG1109) have been identified in the genome of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. We previously found that TK1777 actually encodes a phosphopentomutase. In order to determine which of the remaining three orthologues encodes a phosphoglucomutase (PGM), we examined the PGM activity in T. kodakaraensis cells and identified the gene responsible for this activity. Heterologous gene expression and purification and characterization of the recombinant protein indicated that TK1108 encoded a protein with high levels of PGM activity (690 U mg(-1)), along with high levels of PMM activity (401 U mg(-1)). Similar analyses of the remaining two orthologues revealed that their protein products exhibited neither PGM nor PMM activity. PGM activity and transcription of TK1108 in T. kodakaraensis were found to be higher in cells grown on starch than in cells grown on pyruvate. Our results clearly indicate that, among the four PMM gene orthologues in T. kodakaraensis, only one gene, TK1108, actually encodes a protein with PGM and PMM activities. PMID- 15342578 TI - Biosynthesis of chloro-beta-hydroxytyrosine, a nonproteinogenic amino acid of the peptidic backbone of glycopeptide antibiotics. AB - The role of the putative P450 monooxygenase OxyD and the chlorination time point in the biosynthesis of the glycopeptide antibiotic balhimycin produced by Amycolatopsis balhimycina were analyzed. The oxyD gene is located directly downstream of the bhp (perhydrolase) and bpsD (nonribosomal peptide synthetase D) genes, which are involved in the synthesis of the balhimycin building block beta hydroxytyrosine (beta-HT). Reverse transcriptase experiments revealed that bhp, bpsD, and oxyD form an operon. oxyD was inactivated by an in-frame deletion, and the resulting mutant was unable to produce an active compound. Balhimycin production could be restored (i) by complementation with an oxyD gene, (ii) in cross-feeding studies using A. balhimycina JR1 (a null mutant with a block in the biosynthesis pathway of the building blocks hydroxy- and dihydroxyphenylglycine) as an excretor of the missing precursor, and (iii) by supplementation of beta-HT in the growth medium. These data demonstrated an essential role of OxyD in the formation pathway of this amino acid. Liquid chromatography-electrospray ionization-mass spectrometry analysis indicated the biosynthesis of completely chlorinated balhimycin by the oxyD mutant when culture filtrates were supplemented with nonchlorinated beta-HT. In contrast, supplementation with 3 chloro-beta-HT did not restore balhimycin production. These results indicated that the chlorination time point was later than the stage of free beta-HT, most likely during heptapeptide synthesis. PMID- 15342577 TI - The Bacteroides fragilis pathogenicity island is contained in a putative novel conjugative transposon. AB - The genetic element flanking the Bacteroides fragilis pathogenicity island (BfPAI) in enterotoxigenic B. fragilis (ETBF) strain 86-5443-2-2 and a related genetic element in NCTC 9343 were characterized. The results suggested that these genetic elements are members of a new family of conjugative transposons (CTns) not described previously. These putative CTns, designated CTn86 and CTn9343 for ETBF 86-5443-2-2 and NCTC 9343, respectively, differ from previously described Bacteroides species CTns in a number of ways. These new transposons do not carry tetQ, and the excision from the chromosome to form a circular intermediate is not regulated by tetracycline; they are predicted to differ in their mechanism of transposition; and their sequences have very limited similarity with CTnDOT or other described CTns. CTn9343 is 64,229 bp in length, contains 61 potential open reading frames, and both ends contain IS21 transposases. Colony blot hybridization, PCR, and sequence analysis indicated that CTn86 has the same structure as CTn9343 except that CTn86 lacks a approximately 7-kb region containing truncated integrase (int2) and rteA genes and it contains the BfPAI integrated between the mob region and the bfmC gene. If these putative CTns were to be demonstrated to be transmissible, this would suggest that the bft gene can be transferred from ETBF to nontoxigenic B. fragilis strains by a mechanism similar to that for the spread of antibiotic resistance genes. PMID- 15342579 TI - Regulation of tetralin biodegradation and identification of genes essential for expression of thn operons. AB - The tetralin biodegradation genes of Sphingomonas macrogolitabida strain TFA are clustered in two closely linked and divergent operons. To analyze expression of both operons under different growth conditions, transcriptional and translational gene fusions of the first genes of each operon to lacZ have been constructed in plasmids unable to replicate in Sphingomonas and integrated by recombination into the genome of strain TFA. Expression analysis indicated that the transcription of both genes is induced in similar ways by the presence of tetralin. Gene expression in both operons is also subjected to overimposed catabolic repression. Two additional genes named thnR and thnY have been identified downstream of thnCA3A4 genes. ThnR is similar to LysR-type regulators, and mutational analysis indicated that ThnR is strictly required for expression of the thn operons. Unlike other LysR-type regulators, ThnR does not repress its own synthesis. In fact, ThnR activates its own expression, since thnR is cotranscribed with the thnCA3A4 genes. ThnY is similar to the ferredoxin reductase components of dioxygenase systems and shows the fer2 domain, binding a Cys4[2Fe-2S] iron sulfur center, and the FAD-binding domain, common to those reductases. However, it lacks the NAD-binding domain. Intriguingly, ThnY has a regulatory role, since it is also strictly required for expression of the thn operons. Given the similarity of ThnY to reductases and the possibility of its being present in the two redox states, it is tempting to speculate that ThnY is a regulatory component connecting expression of the thn operons to the physiological status of the cell. PMID- 15342581 TI - Reversible phase variation in the phnE gene, which is required for phosphonate metabolism in Escherichia coli K-12. AB - It is known that Escherichia coli K-12 is cryptic (Phn-) for utilization of methyl phosphonate (MePn) and that Phn+ variants can be selected for growth on MePn as the sole P source. Variants arise from deletion via a possible slip strand mechanism of one of three direct 8-bp repeat sequences in phnE, which restores function to a component of a putative ABC type transporter. Here we show that Phn+ variants are present at the surprisingly high frequency of >10(-2) in K 12 strains. Amplified-fragment length polymorphism analysis was used to monitor instability in phnE in various strains growing under different conditions. This revealed that, once selection for growth on MePn is removed, Phn+ revertants reappear and accumulate at high levels through reinsertion of the 8-bp repeat element sequence. It appears that, in K-12, phnE contains a high-frequency reversible gene switch, producing phase variation which either allows ("on" form) or blocks ("off" form) MePn utilization. The switch can also block usage of other metabolizable alkyl phosphonates, including the naturally occurring 2 aminoethylphosphonate. All K-12 strains, obtained from collections, appear in the "off" form even when bearing mutations in mutS, mutD, or dnaQ which are known to enhance slip strand events between repetitive sequences. The ability to inactivate the phnE gene appears to be unique to K-12 strains since the B strain is naturally Phn+ and lacks the inactivating 8-bp insertion in phnE, as do important pathogenic strains for which genome sequences are known and also strains isolated recently from environmental sources. PMID- 15342580 TI - Structural determinants required to target penicillin-binding protein 3 to the septum of Escherichia coli. AB - In Escherichia coli, cell division is mediated by the concerted action of about 12 proteins that assemble at the division site to presumably form a complex called the divisome. Among these essential division proteins, the multimodular class B penicillin-binding protein 3 (PBP3), which is specifically involved in septal peptidoglycan synthesis, consists of a short intracellular M1-R23 peptide fused to a F24-L39 membrane anchor that is linked via a G40-S70 peptide to an R71 I236 noncatalytic module itself linked to a D237-V577 catalytic penicillin binding module. On the basis of localization analyses of PBP3 mutants fused to green fluorescent protein by fluorescence microscopy, it appears that the first 56 amino acid residues of PBP3 containing the membrane anchor and the G40-E56 peptide contain the structural determinants required to target the protein to the cell division site and that none of the putative protein interaction sites present in the noncatalytic module are essential for the positioning of the protein to the division site. Based on the effects of increasing production of FtsQ or FtsW on the division of cells expressing PBP3 mutants, it is suggested that these proteins could interact. We postulate that FtsQ could play a role in regulating the assembly of these division proteins at the division site and the activity of the peptidoglycan assembly machineries within the divisome. PMID- 15342582 TI - In vivo phosphorylation of partner switching regulators correlates with stress transmission in the environmental signaling pathway of Bacillus subtilis. AB - Exposure of bacteria to diverse growth-limiting stresses induces the synthesis of a common set of proteins which provide broad protection against future, potentially lethal stresses. Among Bacillus subtilis and its relatives, this general stress response is controlled by the sigmaB transcription factor. Signals of environmental and energy stress activate sigmaB through a multicomponent network that functions via a partner switching mechanism, in which protein protein interactions are governed by serine and threonine phosphorylation. Here, we tested a central prediction of the current model for the environmental signaling branch of this network. We used isoelectric focusing and immunoblotting experiments to determine the in vivo phosphorylation states of the RsbRA and RsbS regulators, which act in concert to negatively control the RsbU environmental signaling phosphatase. As predicted by the model, the ratio of the phosphorylated to unphosphorylated forms of both RsbRA and RsbS increased in response to salt or ethanol stress. However, these two regulators differed substantially with regard to the extent of their phosphorylation under both steady-state and stress conditions, with RsbRA always the more highly modified. Mutant analysis showed that the RsbT kinase, which is required for environmental signaling, was also required for the in vivo phosphorylation of RsbRA and RsbS. Moreover, the T171A alteration of RsbRA, which blocks environmental signaling, also blocked in vivo phosphorylation of RsbRA and impeded phosphorylation of RsbS. These in vivo results corroborate previous genetic analyses and link the phosphorylated forms of RsbRA and RsbS to the active transmission of environmental stress signals. PMID- 15342583 TI - An SOS response induced by high pressure in Escherichia coli. AB - Although pressure is an important environmental parameter in microbial niches such as the deep sea and is furthermore used in food preservation to inactivate microorganisms, the fundamental understanding of its effects on bacteria remains fragmentary. Our group recently initiated differential fluorescence induction screening to search for pressure-induced Escherichia coli promoters and has already reported induction of the heat shock regulon. Here the screening was continued, and we report for the first time that pressure induces a bona fide SOS response in E. coli, characterized by the RecA and LexA-dependent expression of uvrA, recA, and sulA. Moreover, it was shown that pressure is capable of triggering lambda prophage induction in E. coli lysogens. The remnant lambdoid e14 element, however, could not be induced by pressure, as opposed to UV irradiation, indicating subtle differences between the pressure-induced and the classical SOS response. Furthermore, the pressure-induced SOS response seems not to be initiated by DNA damage, since DeltarecA and lexA1 (Ind-) mutants, which are intrinsically hypersensitive to DNA damage, were not sensitized or were only very slightly sensitized for pressure-mediated killing and since pressure treatment was not found to be mutagenic. In light of these findings, the current knowledge of pressure-mediated effects on bacteria is discussed. PMID- 15342584 TI - Arginine biosynthesis in Thermotoga maritima: characterization of the arginine sensitive N-acetyl-L-glutamate kinase. AB - To help clarify the control of arginine synthesis in Thermotoga maritima, the putative gene (argB) for N-acetyl-L-glutamate kinase (NAGK) from this microorganism was cloned and overexpressed, and the resulting protein was purified and shown to be a highly thermostable and specific NAGK that is potently and selectively inhibited by arginine. Therefore, NAGK is in T. maritima the feedback control point of arginine synthesis, a process that in this organism involves acetyl group recycling and appears not to involve classical acetylglutamate synthase. The inhibition of NAGK by arginine was found to be pH independent and to depend sigmoidally on the concentration of arginine, with a Hill coefficient (N) of approximately 4, and the 50% inhibitory arginine concentration (I0.5) was shown to increase with temperature, approaching above 65 degrees C the I0.50 observed at 37 degrees C with the mesophilic NAGK of Pseudomonas aeruginosa (the best-studied arginine-inhibitable NAGK). At 75 degrees C, the inhibition by arginine of T. maritima NAGK was due to a large increase in the Km for acetylglutamate triggered by the inhibitor, but at 37 degrees C arginine also substantially decreased the Vmax of the enzyme. The NAGKs of T. maritima and P. aeruginosa behaved in gel filtration as hexamers, justifying the sigmoidicity and high Hill coefficient of arginine inhibition, and arginine or the substrates failed to disaggregate these enzymes. In contrast, Escherichia coli NAGK is not inhibited by arginine and is dimeric, and thus the hexameric architecture may be an important determinant of arginine sensitivity. Potential thermostability determinants of T. maritima NAGK are also discussed. PMID- 15342585 TI - RsbV-independent induction of the SigB-dependent general stress regulon of Bacillus subtilis during growth at high temperature. AB - General stress proteins protect Bacillus subtilis cells against a variety of environmental insults. This adaptive response is particularly important for nongrowing cells, to which it confers a multiple, nonspecific, and preemptive stress resistance. Induction of the general stress response relies on the alternative transcription factor, SigB, whose activity is controlled by a partner switching mechanism that also involves the anti-sigma factor, RsbW, and the antagonist protein, RsbV. Recently, the SigB regulon has been shown to be continuously induced and functionally important in cells actively growing at low temperature. With the exception of this chill induction, all SigB-activating stimuli identified so far trigger a transient expression of the SigB regulon that depends on RsbV. Through a proteome analysis and Northern blot and gene fusion experiments, we now show that the SigB regulon is continuously induced in cells growing actively at 51 degrees C, close to the upper growth limit of B. subtilis. This heat induction of SigB-dependent genes requires the environmental stress responsive phosphatase RsbU, but not the metabolic stress-responsive phosphatase RsbP. RsbU dependence of SigB activation by heat is overcome in mutants that lack RsbV. In addition, loss of RsbV alone or in combination with RsbU triggers a hyperactivation of the general stress regulon exclusively at high temperatures detrimental for cell growth. These new facets of heat induction of the SigB regulon indicate that the current view of the complex genetic and biochemical regulation of SigB activity is still incomplete and that SigB perceives signals independent of the RsbV-mediated signal transduction pathways under heat stress conditions. PMID- 15342586 TI - Characterization of heme uptake cluster genes in the fish pathogen Vibrio anguillarum. AB - Vibrio anguillarum can utilize hemin and hemoglobin as sole iron sources. In previous work we identified HuvA, the V. anguillarum outer membrane heme receptor by complementation of a heme utilization mutant with a cosmid clone (pML1) isolated from a genomic library of V. anguillarum. In the present study, we describe a gene cluster contained in cosmid pML1, coding for nine potential heme uptake and utilization proteins: HuvA, the heme receptor; HuvZ and HuvX; TonB, ExbB, and ExbD; HuvB, the putative periplasmic binding protein; HuvC, the putative inner membrane permease; and HuvD, the putative ABC transporter ATPase. A V. anguillarum strain with an in-frame chromosomal deletion of the nine-gene cluster was impaired for growth with heme or hemoglobin as the sole iron source. Single-gene in-frame deletions were constructed, demonstrating that each of the huvAZBCD genes are essential for utilization of heme as an iron source in V. anguillarum, whereas huvX is not. When expressed in Escherichia coli hemA (strain EB53), a plasmid carrying the gene for the heme receptor, HuvA, was sufficient to allow the use of heme as the porphyrin source. For utilization of heme as an iron source in E. coli ent (strain 101ESD), the tonB exbBD and huvBCD genes were required in addition to huvA. The V. anguillarum heme uptake cluster shows some differences in gene arrangement when compared to homologous clusters described for other Vibrio species. PMID- 15342587 TI - AglZ is a filament-forming coiled-coil protein required for adventurous gliding motility of Myxococcus xanthus. AB - The aglZ gene of Myxococcus xanthus was identified from a yeast two-hybrid assay in which MglA was used as bait. MglA is a 22-kDa cytoplasmic GTPase required for both adventurous and social gliding motility and sporulation. Genetic studies showed that aglZ is part of the A motility system, because disruption or deletion of aglZ abolished movement of isolated cells and aglZ sglK double mutants were nonmotile. The aglZ gene encodes a 153-kDa protein that interacts with purified MglA in vitro. The N terminus of AglZ shows similarity to the receiver domain of two-component response regulator proteins, while the C terminus contains heptad repeats characteristic of coiled-coil proteins, such as myosin. Consistent with this motif, expression of AglZ in Escherichia coli resulted in production of striated lattice structures. Similar to the myosin heavy chain, the purified C terminal coiled-coil domain of AglZ forms filament structures in vitro. PMID- 15342589 TI - New protein-protein interactions identified for the regulatory and structural components and substrates of the type III Secretion system of the phytopathogen Xanthomonas axonopodis Pathovar citri. AB - We have initiated a project to identify protein-protein interactions involved in the pathogenicity of the bacterial plant pathogen Xanthomonas axonopodis pv. citri. Using a yeast two-hybrid system based on Gal4 DNA-binding and activation domains, we have focused on identifying interactions involving subunits, regulators, and substrates of the type III secretion system coded by the hrp (for hypersensitive response and pathogenicity), hrc (for hrp conserved), and hpa (for hrp associated) genes. We have identified several previously uncharacterized interactions involving (i) HrpG, a two-component system response regulator responsible for the expression of X. axonopodis pv. citri hrp operons, and XAC0095, a previously uncharacterized protein encountered only in Xanthomonas spp.; (ii) HpaA, a protein secreted by the type III secretion system, HpaB, and the C-terminal domain of HrcV; (iii) HrpB1, HrpD6, and HrpW; and (iv) HrpB2 and HrcU. Homotropic interactions were also identified for the ATPase HrcN. These newly identified protein-protein interactions increase our understanding of the functional integration of phytopathogen-specific type III secretion system components and suggest new hypotheses regarding the molecular mechanisms underlying Xanthomonas pathogenicity. PMID- 15342588 TI - The small noncoding DsrA RNA is an acid resistance regulator in Escherichia coli. AB - DsrA RNA is a small (87-nucleotide) regulatory RNA of Escherichia coli that acts by RNA-RNA interactions to control translation and turnover of specific mRNAs. Two targets of DsrA regulation are RpoS, the stationary-phase and stress response sigma factor (sigmas), and H-NS, a histone-like nucleoid protein and global transcription repressor. Genes regulated globally by RpoS and H-NS include stress response proteins and virulence factors for pathogenic E. coli. Here, by using transcription profiling via DNA arrays, we have identified genes induced by DsrA. Steady-state levels of mRNAs from many genes increased with DsrA overproduction, including multiple acid resistance genes of E. coli. Quantitative primer extension analysis verified the induction of individual acid resistance genes in the hdeAB, gadAX, and gadBC operons. E. coli K-12 strains, as well as pathogenic E. coli O157:H7, exhibited compromised acid resistance in dsrA mutants. Conversely, overproduction of DsrA from a plasmid rendered the acid-sensitive dsrA mutant extremely acid resistant. Thus, DsrA RNA plays a regulatory role in acid resistance. Whether DsrA targets acid resistance genes directly by base pairing or indirectly via perturbation of RpoS and/or H-NS is not known, but in either event, our results suggest that DsrA RNA may enhance the virulence of pathogenic E. coli. PMID- 15342590 TI - Novel xylose dehydrogenase in the halophilic archaeon Haloarcula marismortui. AB - During growth of the halophilic archaeon Haloarcula marismortui on D-xylose, a specific D-xylose dehydrogenase was induced. The enzyme was purified to homogeneity. It constitutes a homotetramer of about 175 kDa and catalyzed the oxidation of xylose with both NADP+ and NAD+ as cosubstrates with 10-fold higher affinity for NADP+. In addition to D-xylose, D-ribose was oxidized at similar kinetic constants, whereas D-glucose was used with about 70-fold lower catalytic efficiency (kcat/Km). With the N-terminal amino acid sequence of the subunit, an open reading frame (ORF)-coding for a 39.9-kDA protein-was identified in the partially sequenced genome of H. marismortui. The function of the ORF as the gene designated xdh and coding for xylose dehydrogenase was proven by its functional overexpression in Escherichia coli. The recombinant enzyme was reactivated from inclusion bodies following solubilization in urea and refolding in the presence of salts, reduced and oxidized glutathione, and substrates. Xylose dehydrogenase showed the highest sequence similarity to glucose-fructose oxidoreductase from Zymomonas mobilis and other putative bacterial and archaeal oxidoreductases. Activities of xylose isomerase and xylulose kinase, the initial reactions of xylose catabolism of most bacteria, could not be detected in xylose-grown cells of H. marismortui, and the genes that encode them, xylA and xylB, were not found in the genome of H. marismortui. Thus, we propose that this first characterized archaeal xylose dehydrogenase catalyzes the initial step in xylose degradation by H. marismortui. PMID- 15342591 TI - Inactivations of rsbU and sarA by IS256 represent novel mechanisms of biofilm phenotypic variation in Staphylococcus epidermidis. AB - Expression of ica operon-mediated biofilm formation in Staphylococcus epidermidis RP62A is subject to phase variable regulation. Reversible transposition of IS256 into icaADBC or downregulation of icaADBC expression are two important mechanisms of biofilm phenotypic variation. Interestingly, the presence of IS256 was generally associated with a more rapid rate of phenotypic variation, suggesting that IS256 insertions outside the ica locus may affect ica transcription. Consistent with this, we identified variants with diminished ica expression, which were associated with IS256 insertions in the sigmaB activator rsbU or sarA. Biofilm development and ica expression were activated only by ethanol and not NaCl in rsbU::IS256 insertion variants, which were present in approximately 11% of all variants. sigmaB activity was impaired in rsbU::IS256 variants, as evidenced by reduced expression of the sigmaB-regulated genes asp23, csb9, and rsbV. Moreover, expression of sarA, which is sigmaB regulated, and SarA-regulated RNAIII were also suppressed. A biofilm-forming phenotype was restored to rsbU::IS256 variants only after repeated passage and was not associated with IS256 excision from rsbU. Only one sarA::IS256 insertion mutant was identified among 43 biofilm-negative variants. Both NaCl and ethanol-activated ica expression in this sarA::IS256 variant, but only ethanol increased biofilm development. Unlike rsbU::IS256 variants, reversion of the sarA::IS256 variant to a biofilm-positive phenotype was accompanied by precise excision of IS256 from sarA and restoration of normal ica expression. These data identify new roles for IS256 in ica and biofilm phenotypic variation and demonstrate the capacity of this element to influence the global regulation of transcription in S. epidermidis. PMID- 15342592 TI - The hFbpABC transporter from Haemophilus influenzae functions as a binding protein-dependent ABC transporter with high specificity and affinity for ferric iron. AB - Pathogenic Haemophilus influenzae, Neisseria spp. (Neisseria gonorrhoeae and N. meningitidis), Serratia marcescens, and other gram-negative bacteria utilize a periplasm-to-cytosol FbpABC iron transporter. In this study, we investigated the H. influenzae FbpABC transporter in a siderophore-deficient Escherichia coli background to assess biochemical aspects of FbpABC transporter function. Using a radiolabeled Fe3+ transport assay, we established an apparent Km=0.9 microM and Vmax=1.8 pmol/10(7)cells/min for FbpABC-mediated transport. Complementation experiments showed that hFbpABC is dependent on the FbpA binding protein for transport. The ATPase inhibitor sodium orthovanadate demonstrated dose-dependent inhibition of FbpABC transport, while the protonmotive-force-inhibitor carbonyl cyanide m-chlorophenyl hydrazone had no effect. Metal competition experiments demonstrated that the transporter has high specificity for Fe3+ and selectivity for trivalent metals, including Ga3+ and Al3+, over divalent metals. Metal sensitivity experiments showed that several divalent metals, including copper, nickel, and zinc, exhibited general toxicity towards E. coli. Significantly, gallium-induced toxicity was specific only to E. coli expressing FbpABC. A single amino-acid mutation in the gene encoding the periplasmic binding protein, FbpA(Y196I), resulted in a greatly diminished iron binding affinity Kd=5.2 x 10( 4) M(-1), approximately 14 orders of magnitude weaker than that of the wild-type protein. Surprisingly, the mutant transporter [FbpA(Y196I)BC] exhibited substantial transport activity, approximately 35% of wild-type transport, with Km=1.2 microM and Vmax=0.5 pmol/10(7)cells/min. We conclude that the FbpABC complexes possess basic characteristics representative of the family of bacterial binding protein-dependent ABC transporters. However, the specificity and high affinity binding characteristics suggest that the FbpABC transporters function as specialized transporters satisfying the strict chemical requirements of ferric iron (Fe3+) binding and membrane transport. PMID- 15342593 TI - Bacillus subtilis StoA Is a thiol-disulfide oxidoreductase important for spore cortex synthesis. AB - Bacillus subtilis is an endospore-forming bacterium. There are indications that protein disulfide linkages occur in spores, but the role of thiol-disulfide chemistry in spore synthesis is not understood. Thiol-disulfide oxidoreductases catalyze formation or breakage of disulfide bonds in proteins. CcdA is the only B. subtilis thiol-disulfide oxidoreductase that has previously been shown to play some role in endospore biogenesis. In this work we show that lack of the StoA (YkvV) protein results in spores sensitive to heat, lysozyme, and chloroform. Compared to CcdA deficiency, StoA deficiency results in a 100-fold-stronger negative effect on sporulation efficiency. StoA is a membrane-bound protein with a predicted thioredoxin-like domain probably localized in the intermembrane space of the forespore. Electron microscopy of spores of CcdA- and StoA-deficient strains showed that the spore cortex is absent in both cases. The BdbD protein catalyzes formation of disulfide bonds in proteins on the outer side of the cytoplasmic membrane but is not required for sporulation. Inactivation of bdbD was found to suppress the sporulation defect of a strain deficient in StoA. Our results indicate that StoA is a thiol-disulfide oxidoreductase that is involved in breaking disulfide bonds in cortex components or in proteins important for cortex synthesis. PMID- 15342595 TI - Complementation of an Escherichia coli DnaK defect by Hsc70-DnaK chimeric proteins. AB - Escherichia coli DnaK and rat Hsc70 are members of the highly conserved 70-kDa heat shock protein (Hsp70) family that show strong sequence and structure similarities and comparable functional properties in terms of interactions with peptides and unfolded proteins and cooperation with cochaperones. We show here that, while the DnaK protein is, as expected, able to complement an E. coli dnaK mutant strain for growth at high temperatures and lambda phage propagation, Hsc70 protein is not. However, an Hsc70 in which the peptide-binding domain has been replaced by that of DnaK is able to complement this strain for both phenotypes, suggesting that the peptide-binding domain of DnaK is essential to fulfill the specific functions of this protein necessary for growth at high temperatures and for lambda phage replication. The implications of these findings on the functional specificities of the Hsp70s and the role of protein-protein interactions in the DnaK chaperone system are discussed. PMID- 15342594 TI - Mutational analysis of Xanthomonas harpin HpaG identifies a key functional region that elicits the hypersensitive response in nonhost plants. AB - HpaG is a type III-secreted elicitor protein of Xanthomonas axonopodis pv. glycines. We have determined the critical amino acid residues important for hypersensitive response (HR) elicitation by random and site-directed mutagenesis of HpaG and its homolog XopA. A plasmid clone carrying hpaG was mutagenized by site-directed mutagenesis, hydroxylamine mutagenesis, and error-prone PCR. A total of 52 mutants were obtained, including 51 single missense mutants and 1 double missense mutant. The HR elicitation activity was abolished in the two missense mutants [HpaG(L50P) and HpaG(L43P/L50P)]. Seven single missense mutants showed reduced activity, and the HR elicitation activity of the rest of the mutants was similar to that of wild-type HpaG. Mutational and deletion analyses narrowed the region essential for elicitor activity to the 23-amino-acid peptide (H2N-NQGISEKQLDQLLTQLIMALLQQ-COOH). A synthetic peptide of this sequence possessed HR elicitor activity at the same concentration as the HpaG protein. This region has 78 and 74% homology with 23- and 27-amino-acid regions of the HrpW harpin domains, respectively, from Pseudomonas and Erwinia spp. The secondary structure of the peptide is predicted to be an alpha-helix, as is the HrpW region that is homologous to HpaG. The predicted alpha-helix of HpaG is probably critical for the elicitation of the HR in tobacco plants. In addition, mutagenesis of a xopA gene yielded two gain-of-function mutants: XopA(F48L) and XopA(F48L/M52L). These results indicate that the 12 amino acid residues between L39 and L50 of HpaG have critical roles in HR elicitation in tobacco plants. PMID- 15342596 TI - Transcriptional Repressor CopR: use of SELEX to study the copR operator indicates that evolution was directed at maximal binding affinity. AB - CopR is one of the two copy number control elements of the streptococcal plasmid pIP501. It represses transcription of the repR mRNA encoding the essential replication initiator protein about 10- to 20-fold by binding to its operator region upstream of the repR promoter pII. CopR binds at two consecutive sites in the major groove of the DNA that share the consensus motif 5'-CGTG. Previously, the minimal operator was narrowed down to 17 bp, and equilibrium dissociation constants for DNA binding and dimerization were determined to be 0.4 nM and 1.4 microM, respectively. In this work, we used a SELEX procedure to study copR operator sequences of different lengths in combination with electrophoretic mobility shift assays of mutated copR operators as well as copy number determinations to assess the sequence requirements for CopR binding. The results suggest that in vivo evolution was directed at maximal binding affinity. Three simultaneous nucleotide exchanges outside the bases directly contacted by CopR only slightly affected CopR binding in vitro or copy numbers in vivo. Furthermore, the optimal spacer sequence was found to comprise 7 bp, to be AT rich, and to need an A/T and a T at the 3' positions, whereas broad variations in the sequences flanking the minimal 17-bp operator were well tolerated. PMID- 15342602 TI - mRNA secondary structure modulates translation of Tat-dependent formate dehydrogenase N. AB - Formate dehydrogenase N (FDH-N) of Escherichia coli is a membrane-bound enzyme comprising FdnG, FdnH, and FdnI subunits organized in an (alphabetagamma)3 configuration. The FdnG subunit carries a Tat-dependent signal peptide, which localizes the protein complex to the periplasmic side of the membrane. We noted that substitution of the first arginine (R5) in the twin arginine signal sequence of FdnG for a variety of other amino acids resulted in a dramatic (up to 60-fold) increase in the levels of protein synthesized. Bioinformatic analysis suggested that the mRNA specifying the first 17 codons of fdnG forms a stable stem-loop structure. A detailed mutational analysis has demonstrated the importance of this mRNA stem-loop in modulating FDH-N translation. PMID- 15342603 TI - Bacterial thymidylate synthase with intein, group II Intron, and distinctive ThyX motifs. AB - The ThyX class of thymidylate synthases was previously characterized by a common ThyX motif, RHRX7S. We report bacterial ThyX sequences having distinctive ThyX motifs, suggesting a more general ThyX motif, R/THRX7-8S. One ThyX sequence has an intein in its ThyX motif that was shown to do protein splicing and a group II intron in its gene, suggesting a hot spot for these self-splicing mobile elements. PMID- 15342601 TI - Transcription by Methanothermobacter thermautotrophicus RNA polymerase in vitro releases archaeal transcription factor B but not TATA-box binding protein from the template DNA. AB - Transcription initiation in Archaea requires the assembly of a preinitiation complex containing the TATA- box binding protein (TBP), transcription factor B (TFB), and RNA polymerase (RNAP). The results reported establish the fate of Methanothermobacter thermautotrophicus TBP and TFB following transcription initiation by M. thermautotrophicus RNAP in vitro. TFB is released after initiation, during extension of the transcript from 4 to 24 nucleotides, but TBP remains bound to the template DNA. Regulation of archaeal transcription initiation by a repressor competition with TBP for TATA-box region binding must accommodate this observation. PMID- 15342598 TI - Amino acid contacts between sigma 70 domain 4 and the transcription activators RhaS and RhaR. AB - The RhaS and RhaR proteins are transcription activators that respond to the availability of L-rhamnose and activate transcription of the operons in the Escherichia coli L-rhamnose catabolic regulon. RhaR activates transcription of rhaSR, and RhaS activates transcription of the operon that encodes the L-rhamnose catabolic enzymes, rhaBAD, as well as the operon that encodes the L-rhamnose transport protein, rhaT. RhaS is 30% identical to RhaR at the amino acid level, and both are members of the AraC/XylS family of transcription activators. The RhaS and RhaR binding sites overlap the -35 hexamers of the promoters they regulate, suggesting they may contact the sigma70 subunit of RNA polymerase as part of their mechanisms of transcription activation. In support of this hypothesis, our lab previously identified an interaction between RhaS residue D241 and sigma70 residue R599. In the present study, we first identified two positively charged amino acids in sigma70, K593 and R599, and three negatively charged amino acids in RhaR, D276, E284, and D285, that were important for RhaR mediated transcription activation of the rhaSR operon. Using a genetic loss-of contact approach we have obtained evidence for a specific contact between RhaR D276 and sigma70 R599. Finally, previous results from our lab separately showed that RhaS D250A and sigma70 K593A were defective at the rhaBAD promoter. Our genetic loss-of-contact analysis of these residues indicates that they identify a second site of contact between RhaS and sigma70. PMID- 15342597 TI - A novel mutation within the central Listeria monocytogenes regulator PrfA that results in constitutive expression of virulence gene products. AB - The PrfA protein of Listeria monocytogenes functions as a key regulatory factor for the coordinated expression of many virulence genes during bacterial infection of host cells. PrfA activity is controlled by multiple regulatory mechanisms, including an apparent requirement for either the presence of a cofactor or some form of posttranslational modification that regulates the activation of PrfA. In this study, we describe the identification and characterization of a novel PrfA mutation that results in constitutive activation of the PrfA protein. The PrfA L140F mutation was found to confer high-level expression of PrfA-regulated genes and to be functionally dominant over the wild-type allele. The presence of the PrfA L140F mutation resulted in the aggregation of L. monocytogenes in broth culture and, unlike previously described prfA mutations, appeared to be slightly toxic to the bacteria. High-level PrfA-dependent gene expression showed no additional increase in L. monocytogenes strains containing an additional copy of prfA L140F despite a >4-fold increase in PrfA protein levels. In contrast, the introduction of multiple copies of the wild-type prfA allele to L. monocytogenes resulted in a corresponding increase in PrfA-dependent gene expression, although overall expression levels remained far below those observed for PrfA L140F strains. These results suggest a hierarchy of PrfA regulation, such that the relative levels of PrfA protein present within the cell correlate with the levels of PrfA-dependent gene expression when the protein is not in its fully activated state; however, saturating levels of the protein are then quickly reached when PrfA is converted to its active form. Regulation of the PrfA activation status must be an important facet of L. monocytogenes survival, as mutations that result in constitutive PrfA activation may have deleterious consequences for bacterial physiology. PMID- 15342605 TI - Recombinant cyclophilins lack nuclease activity. AB - Several single-domain prokaryotic and eukaryotic cyclophilins have been identified as also being unspecific nucleases with a role in DNA degradation during the lytic processes that accompany bacterial cell death and eukaryotic apoptosis. Evidence is provided here that the supposed nuclease activity of human and bacterial recombinant cyclophilins is due to contamination of the proteins by the host Escherichia coli endonuclease and is not an intrinsic property of these proteins. PMID- 15342599 TI - The paralogous pairs of genes involved in clavulanic acid and clavam metabolite biosynthesis are differently regulated in Streptomyces clavuligerus. AB - Carboxyethylarginine synthase, encoded by the paralogous ceaS1 and ceaS2 genes, catalyzes the first reaction in the shared biosynthetic pathway leading to clavulanic acid and the other clavam metabolites in Streptomyces clavuligerus. The nutritional regulation of ceaS1 and ceaS2 expression was analyzed by reverse transcriptase PCR and by the use of the enhanced green fluorescent protein encoding gene (egfp) as a reporter. ceaS1 was transcribed in complex soy medium only, whereas ceaS2 was transcribed in both soy and defined starch-asparagine (SA) media. The transcriptional start points of the two genes were also mapped to a C residue 98 bp upstream of ceaS1 and a G residue 51 bp upstream of the ceaS2 start codon by S1 nuclease protection and primer extension analyses. Furthermore, transcriptional mapping of the genes encoding the beta-lactam synthetase (bls1) and proclavaminate amidinohydrolase (pah1) isoenzymes from the paralogue gene cluster indicated that a single polycistronic transcript of approximately 4.9 kb includes ceaS1, bls1, and pah1. The expression of ceaS1 and ceaS2 in a mutant strain defective in the regulatory protein CcaR was also examined. ceaS1 transcription was not affected in the ccaR mutant, whereas that of ceaS2 was greatly reduced compared to the wild-type strain. Overall, our results suggest that different mechanisms are involved in regulating the expression of ceaS1 and ceaS2, and presumably also of other paralogous genes that encode proteins involved in the early stages of clavulanic acid and clavam metabolite biosynthesis. PMID- 15342600 TI - Temporal global changes in gene expression during temperature transition in Yersinia pestis. AB - DNA microarrays encompassing the entire genome of Yersinia pestis were used to characterize global regulatory changes during steady-state vegetative growth occurring after shift from 26 to 37 degrees C in the presence and absence of Ca2+. Transcriptional profiles revealed that 51, 4, and 13 respective genes and open reading frames (ORFs) on pCD, pPCP, and pMT were thermoinduced and that the majority of these genes carried by pCD were downregulated by Ca2+. In contrast, Ca2+ had little effect on chromosomal genes and ORFs, of which 235 were thermally upregulated and 274 were thermally downregulated. The primary consequence of these regulatory events is profligate catabolism of numerous metabolites available in the mammalian host. PMID- 15342604 TI - The secretion signal of YopN, a regulatory protein of the Yersinia enterocolitica type III secretion pathway. AB - The type III secretion signal of Yersinia enterocolitica YopN was mapped using a gene fusion approach. yopN codons 1 to 12 were identified as critical for signal function. Several synonymous mutations that abolish secretion of hybrid proteins without altering the codon specificity of yopN mRNA were identified. PMID- 15342608 TI - Processing of the tail lysozyme (gp5) of bacteriophage T4. AB - The processing site of gp5 has been determined to be between residues Val-390 and His-391, instead of Ser-351 and Ala-352 as previously reported (H. Kanamaru, N. C. Gassner, N. Ye, S. Takeda, and F. Arisaka, J. Bacteriol. 181:2739-2744). Moreover, the maturation of gp5 is abolished by null mutations in other hub genes, indicating that cleavage requires the interactions of several baseplate proteins. PMID- 15342607 TI - Extensive genomic polymorphism within Mycobacterium avium. AB - We have initiated comparative genomic analysis of Mycobacterium avium subspecies by DNA microarray, uncovering 14 large sequence polymorphisms (LSPs) comprising over 700 kb that distinguish M. avium subsp. avium from M. avium subsp. paratuberculosis. Genes predicted to encode metabolic pathways were overrepresented in the LSPs, and analysis revealed a polymorphism within the mycobactin biosynthesis operon that potentially explains the in vitro mycobactin dependence of M. avium subsp. paratuberculosis. PMID- 15342606 TI - Biofilm mode of growth of Streptococcus intermedius favored by a competence stimulating signaling peptide. AB - Gram-positive and gram-negative bacteria use quorum sensing to coordinate population behavior. In several streptococci, quorum sensing mediated by competence-stimulating peptides (CSP) is associated with development of competence for transformation. We show here that a synthetic CSP favored the biofilm mode of growth of Streptococcus intermedius without affecting the rate of culture growth. PMID- 15342609 TI - A DNA adenine methyltransferase of Escherichia coli that is cell cycle regulated and essential for viability. PMID- 15342610 TI - Differences in drug pharmacokinetics between East Asians and Caucasians and the role of genetic polymorphisms. AB - Interethnic variability in pharmacokinetics can cause unexpected outcomes such as therapeutic failure, adverse effects, and toxicity in subjects of different ethnic origin undergoing medical treatment. It is important to realize that both genetic and environmental factors can lead to these differences among ethnic groups. The International Conference on Harmonization (ICH) published a guidance to facilitate the registration of drugs among ICH regions (European Union, Japan, the United States) by recommending a framework for evaluating the impact of ethnic factors on a drug's effect, as well as its efficacy and safety at a particular dosage and dosage regimen. This review focuses on the pharmacokinetic differences between East Asians and Caucasians. Differences in metabolism between East Asians and Caucasians are common, especially in the activity of several phase I enzymes such as CYP2D6 and the CYP2C subfamily. Before drug therapy, identification of either the genotype and/or the phenotype for these enzymes may be of therapeutic value, particularly for drugs with a narrow therapeutic index. Furthermore, these differences are relevant for international drug approval when regulatory agencies must decide if they accept results from clinical trials performed in other parts of the world. PMID- 15342611 TI - Could the WHO model list of essential medicines do more for the safe and appropriate use of injections? AB - A national drug policy addressing the safe and appropriate use of injections is an important element to prevent overuse and unsafe use of injections. Because the World Health Organization World Health Organization Model List of Essential Medicines is a keystone of national drug policies, the authors examined the way it addresses injection practices. They reviewed the 11th World Health Organization Model List of Essential Medicines to collect information on (1) injectable medicines, (2) diluents, and (3) the recommendations regarding the procurement of injection devices. Of 306 active ingredients on the list, 135 (44%) are mentioned in injectable form. Of these, 41 (30%) need diluents for reconstitution. The list does not mention the need to procure appropriate diluents, injection devices, and safety boxes in quantities that match the quantities of injectable medicines. In addition, the list provides limited information that can be used to forecast the needs of injection devices to administer the injectable medicines that are included in the list. Future revisions of the World Health Organization Model List of Essential Medicines should attempt to reduce the number of injectable formulations on the basis of evidence. In addition, the list should specify that when injectable medicines are being supplied, diluents, single-use syringes, and safety boxes should be supplied. The volume of syringes needed for administration should be specified for each injectable medication on the list to facilitate the forecasting of the needs of injection devices. PMID- 15342612 TI - Population pharmacokinetics of ibuprofen enantiomers in very premature neonates. AB - The objective of the present study was to evaluate the pharmacokinetic parameters for both S- and R-ibuprofen enantiomers in very premature neonates (gestational age strictly inferior to 28 weeks) and possible relationships between the pharmacokinetic parameters and various covariates. Newborns were randomized to receive ibuprofen or placebo for the prophylactic treatment of patent ductus arteriosus (PDA) at an initial dose of 10 mg/kg ibuprofen within 6 hours after birth, followed by two 5-mg/kg doses at 24-hour intervals (n = 52). If a PDA was still present afterwards, a curative course of ibuprofen using the same dosage regimen was administered (n = 10). A sparse sampling strategy was used because only 2 samples were collected after the third prophylactic injection and 1 after the third curative injection. A model including the chiral transformation of R- to S-ibuprofen was fitted to the concentration-time data using a population approach (NONMEM). R- and S-ibuprofen t(1/2) were about 10 hours and 25.5 hours, respectively. After prophylactic treatment, the mean clearance of R-ibuprofen (CLR = 12.7 mL/h) was about 2.5-fold higher than for S-ibuprofen (CLS = 5.0 mL/h). In addition, clearance of R- and S-ibuprofen increased significantly with gestational age. The mean estimation of R-ibuprofen clearance was found to be higher than for S-ibuprofen, and the clearance of both enantiomers increased with gestational age. This should be considered to assess pharmacokinetic pharmacodynamic relationships of ibuprofen in premature neonates and subsequently to understand and refine the use of ibuprofen in managing PDA either as a prophylactic or curative treatment. PMID- 15342613 TI - The effects of modifying in vivo cytochrome P450 3A (CYP3A) activity on etoricoxib pharmacokinetics and of etoricoxib administration on CYP3A activity. AB - To investigate the influence of modifying in vivo cytochrome P450 3A (CYP3A) activity on the pharmacokinetics of etoricoxib, a selective inhibitor of cyclooxygenase-2, and of etoricoxib administration on CYP3A activity, a 3-part, randomized, crossover study was conducted in 3 panels of healthy volunteers. In part I, 8 subjects were administered a single dose of 60 mg etoricoxib alone and following daily doses of 400 mg ketoconazole, a known strong inhibitor of CYP3A. In part II, 8 different subjects were administered a single dose of 60 mg etoricoxib alone and following daily doses of 600 mg rifampin, a known strong inducer of CYP3A. In parts I and II, plasma samples were collected following each etoricoxib dose and analyzed for etoricoxib. In part III, 8 different subjects were administered 120 mg etoricoxib or placebo once daily for 11 days, and the erythromycin breath test was administered on day 11 of each period. Coadministration of etoricoxib with daily doses of ketoconazole resulted in an average 43% increase in etoricoxib AUC; based on previous studies, this increase would not be expected to have any clinically meaningful effect. In contrast, coadministration of etoricoxib with daily doses of rifampin had a potentially clinically important effect on etoricoxib pharmacokinetics (average 65% decrease in etoricoxib AUC). Etoricoxib had no effect on hepatic CYP3A activity, as assessed by the erythromycin breath test. PMID- 15342614 TI - Pharmacokinetics of dextromethorphan after single or multiple dosing in combination with quinidine in extensive and poor metabolizers. AB - Dextromethorphan (DM) pharmacological properties predict that the widely used cough suppressant could be used to treat several neuronal disorders, but it is rapidly metabolized after oral dosing. To find out whether quinidine (Q), a CYP2D6 inhibitor, could elevate and prolong DM plasma profiles, 2 multiple-dose studies identified the lowest oral dose of Q that could be used in a fixed combination with 3 doses of DM. A multiple-dose study in healthy subjects with an extensive or a poor enzyme metabolizer phenotype evaluated the safety and pharmacokinetic profile of a selected fixed-dose combination (AVP-923). Study 1 randomized 46 healthy subjects, who were extensive CYP2D6 metabolizers, to receive 0, 2.5, 10, 25, 50, or 75 mg Q twice daily in combination with 30 mg DM for 7 days. Plasma and urine samples were collected after the first and last doses for the assay of DM, dextrorphan (DX), and Q. Study 2 randomized 65 healthy extensive CYP2D6 metabolizers to 8 groups given twice-daily 45- or 60-mg DM doses combined with 0, 30, 45, or 60 mg Q for 7 days. The effects of increasing Q were not different with doses greater than 25 mg, whereas lower doses showed a dose related increase in plasma DM concentrations. Urinary ratios of DM/DX showed a Q dose- and time-related increase in the number of subjects converted to the poor metabolizer phenotype that reached 100% on day 3 of dosing with 25 mg Q. Results from both studies indicated that 25 to 30 mg Q is adequate to maximally suppress O-demethylation of DM. Study 3 evaluated 7 extensive metabolizers and 2 poor metabolizers given an oral capsule every 12 hours containing 30 mg Q combined with 30 mg DM. DM plasma AUC values increased in both groups of subjects during the 8-day study. The mean urinary metabolic ratio (DM/DX) increased at least 27 fold in extensive metabolizers by day 8. There was no effect of Q on urinary metabolic ratios in poor metabolizers. Safety evaluations, including electrocardiograms, indicated that the combination was well tolerated, with no difference between extensive and poor metabolizer phenotypes. PMID- 15342615 TI - Pharmacodynamics and pharmacokinetics of oral levosimendan and its metabolites in patients with severe congestive heart failure: a dosing interval study. AB - The objective of this study was to explore the pharmacodynamics and pharmacokinetics of oral levosimendan in patients with severe congestive heart failure. This was a randomized, parallel-group, double-blind, placebo-controlled trial. Oral levosimendan 2 to 8 mg daily or placebo was administered to 25 patients with New York Heart Association class III-IV congestive heart failure for 4 weeks. Pharmacodynamic variables consisted of heart rate-corrected electromechanical systole, heart rate, and systolic and diastolic blood pressure. The pharmacokinetics of levosimendan and its metabolites, OR-1855 and OR-1896, was assessed. The 4- to 8-mg daily doses of oral levosimendan showed moderate inotropic effects. Blood pressure remained unchanged with all doses. A moderate increase in heart rate was observed except with the 2-mg dose. Pharmacokinetic parameters of the metabolites increased linearly with the dose (P < or = .002 for Cmax and AUC0-8h for both treatment groups). It was concluded that oral levosimendan has inotropic and chronotropic effects in patients with severe congestive heart failure. Plasma concentrations of its metabolites increase dose dependently. PMID- 15342616 TI - Dermal absorption of camphor, menthol, and methyl salicylate in humans. AB - Camphor, menthol, and methyl salicylate occur in numerous over-the-counter products. Although extensively used, there have been no estimates of human exposure following administration via dermal application. Furthermore, there is little information about the pharmacokinetics of those compounds. The authors report the plasma concentrations of the intact compounds as a function of dose following dermal patch application. Three groups of 8 subjects (4 male, 4 female) applied a different number of commercial patches (2, 4, or 8) to the skin for 8 hours. Plasma samples were assayed using sensitive and selective gas chromatographic methods. For the 8-patch group, the average maximum plasma concentrations (Cmax +/- SD) were 41.0 +/- 5.8 ng/mL, 31.9 +/- 8.8 ng/mL, and 29.5 +/- 10.5 ng/mL for camphor, menthol, and methyl salicylate, respectively. The corresponding values for the 4-patch group were 26.8 +/- 7.2 ng/mL, 19.0 +/- 5.4 ng/mL, and 16.8 +/- 6.8 ng/mL. The harmonic mean terminal half-lives were 5.6 +/- 1.3 hours, 4.7 +/- 1.6 hours, and 3.0 +/- 1.2 hours for camphor, menthol, and methyl salicylate, respectively. The 2-patch group had measurable but low plasma concentrations of each compound. Low-dose dermal application for an extended time results in low plasma concentrations of all 3 compounds. Four and 8 patches, when applied for 8 hours, gave measurable and nearly proportional plasma concentrations. Although unable to determine the absolute dermal bioavailability of these compounds, there appears to be relatively low systemic exposure to these potentially toxic compounds, even when an unrealistically large number of patches are applied for an unusually long time. PMID- 15342617 TI - Pharmacokinetics of frovatriptan in adolescent migraineurs. AB - Frovatriptan is a selective 5-HT(1B/1D) receptor agonist available for acute treatment of migraine in adults (18 years and older). The objective of this study was to determine key pharmacokinetic parameters of frovatriptan in adolescent migraineurs after a single 2.5-mg oral dose and to compare these results with those from an earlier study completed in adults. Subjects were stratified by age (12-14 and 15-17 years) and gender, and serial blood and urine samples were collected over 48 hours. A total of 25 subjects (13 male, 12 female) completed the study. Pharmacokinetic profiles for adolescent subjects were similar to those observed in adults. The median tmax ranged from 2 (male subjects) to 3 (female subjects) hours. The AUC0-24h and Cmax were slightly lower in adolescent subjects as compared with adults. As seen in adults, AUC0-24h and Cmax values were approximately 2-fold higher in females than in their male counterparts (AUC mean range 40.5-59.8 ng x h/mL vs 21.2-23.5 ng x h/mL and Cmax mean range 4.02-6.14 vs ng x h/mL 2.52-2.99 ng/mL, in female and male adolescent subjects, respectively). Elimination was biphasic, with an approximate terminal elimination half-life (t(1/2)) between 12.2 and 25.5 hours. Renal clearance was similar in adolescents and adults, being somewhat higher in female than male subjects. Frovatriptan was well tolerated with no serious or treatment-related adverse effects. In addition, there were no clinically significant changes in safety parameters. Overall, the pharmacokinetic profile of frovatriptan in adolescents (12-17 years) is similar to that seen in adults, and dosing adjustments are unlikely to be needed. PMID- 15342618 TI - Hand-foot syndrome in patients treated with capecitabine-containing combination chemotherapy. AB - Clinical characteristics and risk factors of hand-foot syndrome were investigated in patients who received capecitabine-containing chemotherapy. Toxicity data were analyzed from 179 patients in 4 prospective clinical trials testing docetaxel/capecitabine/cisplatin in stomach cancer, capecitabine/cisplatin in biliary or stomach cancer, and vinorelbine/capecitabine in breast cancer. Hand foot syndrome was reported in 116/179 (64.8%) of patients, with grade 3 hand-foot syndrome in 8/179 (4.5%). Hand-foot syndrome first developed within the first 3 chemotherapy cycles in 100/116 (86.2%) patients, with the median onset for all 3 treatment regimens occurring during cycle 2. Because severe reactions were rare, hand-foot syndrome was not a major factor influencing treatment schedule. Risk factor analyses showed that combined use of docetaxel and preceding chemotherapy related stomatitis were significant risk factors for the development of hand-foot syndrome. Our results suggest that a combined treatment agent and a patient's susceptibility to chemotherapy-related toxicity may increase the risk of capecitabine-induced hand-foot syndrome. PMID- 15342619 TI - Intramuscular haloperidol or lorazepam and QT intervals in schizophrenia. AB - The objective of this study was to estimate the effects of intramuscular haloperidol and lorazepam on the QT interval in volunteers with schizophrenia. Intramuscular haloperidol and intramuscular lorazepam are standard treatments in the acute management of agitation and aggression. Although prolongation of the QT interval and sequelae, including torsade de pointes and death, have been reported for haloperidol (but not lorazepam), formal studies have been lacking. Volunteers with schizophrenia (n = 12) were administered a single intramuscular injection of 7.5 mg haloperidol or 4 mg lorazepam in a blinded, randomized, placebo-controlled crossover design. Serial EKGs and concurrent blood samples were obtained over 6 hours following each injection. Changes in the QT interval were evaluated, as were plasma drug and prolactin concentrations. Haloperidol injection increased the heart rate-corrected QT interval an average of 5.1 msec using Bazett's correction (QTb 90% confidence interval [CI]: 0.3, 9.8), 3.6 msec using Fridericia's correction (QTf 90% CI: 0.02, 7.2), and 4.2 msec using an empirically derived "baseline correction" (QT(ii) 90% CI: 0.3, 8.0). Effects of lorazepam on QT were nullified by correction for the heart rate elevation (QTb 3.8 msec, 90% CI: 0.6, 7.1; QTf 0.0 msec, 90% CI: -3.2, 3.4; QTii -2.3 msec, 90% CI: -6.6, 2.0). An association between QT prolongation and occurrence of extrapyramidal symptoms was observed. On average, intramuscular haloperidol led to minimal prolongation of the QT interval. This propensity is of theoretical concern in individuals with risk factors for torsade de pointes but seems unlikely to be a problem in the vast majority of patients. PMID- 15342621 TI - N-glucosylation of cytokinins by glycosyltransferases of Arabidopsis thaliana. AB - Cytokinins are plant hormones that can be glucosylated to form O-glucosides and N glucosides. The glycoconjugates are inactive and are thought to play a role in homeostasis of the hormones. Although O-glucosyltransferases have been identified that recognize cytokinins, the enzymes involved in N-glucosylation have not been identified even though the process has been recognized for many years. This study utilizes a screening strategy in which 105 recombinant glycosyltransferases (UGTs) of Arabidopsis have been analyzed for catalytic activity toward the classical cytokinins: trans-zeatin, dihydrozeatin, N(6)-benzyladenine, N(6) isopentenyladenine, and kinetin. Five UGTs were identified in the screen. UGT76C1 and UGT76C2 recognized all cytokinins and glucosylated the hormones at the N(7) and N(9) positions. UGT85A1, UGT73C5, and UGT73C1 recognized trans-zeatin and dihydrozeatin, which have an available hydroxyl group for glucosylation and formed the O-glucosides. The biochemical characteristics of the N glucosyltransferases were analyzed, and highly effective inhibitors of their activities were identified. Constitutive overexpression of UGT76C1 in transgenic Arabidopsis confirmed that the recombinant enzyme functioned in vivo to glucosylate cytokinin applied to the plant. The role of the N glucosyltransferases in cytokinin metabolism is discussed. PMID- 15342622 TI - The stress kinase MRK contributes to regulation of DNA damage checkpoints through a p38gamma-independent pathway. AB - DNA damage induced by ionizing radiation (IR) activates a complex cellular response that includes checkpoints leading to cell cycle arrest. The stress activated mitogen-activated protein kinase (MAPK) p38gamma has been implicated in the G(2) phase checkpoint induced by IR. We recently discovered MRK as a member of the MAPK kinase kinase family that activates p38gamma. Here we investigated the role of MRK in the checkpoint response to IR. We identified autophosphorylation sites on MRK that are important for its kinase activity. A phosphospecific antibody that recognizes these sites showed that MRK is activated upon IR in a rapid and sustained manner. MRK depletion by RNA interference resulted in defective S and G(2) checkpoints induced by IR that were accompanied by reduced Chk2 phosphorylation and delayed Cdc25A degradation. We also showed that Chk2 is a substrate for MRK in vitro and is phosphorylated at Thr(68) by active MRK in cells. MRK depletion also increased sensitivity to the killing effects of IR. In addition, MRK depletion reduced IR-induced activation of p38gamma but had no effect on p38alpha activation, indicating that MRK is a specific activator of p38gamma after IR. Inhibition of p38gamma by RNA interference, however, did not impair IR-induced checkpoints. Thus, in response to IR MRK controls two independent pathways: the Chk2-Cdc25A pathway leading to cell cycle arrest and the p38gamma MAPK pathway. PMID- 15342623 TI - 3,3',5-Triiodo-L-thyronine up-regulation of Na,K-ATPase activity and cell surface expression in alveolar epithelial cells is Src kinase- and phosphoinositide 3 kinase-dependent. AB - We previously reported that thyroid hormone, 3,3',5-triiodo-l-thyronine (T3), increased Na,K-ATPase activity of adult rat alveolar epithelial cells in a transcription-independent manner via increased cell surface expression of the alpha(1) and beta(1) subunits of Na,K-ATPase. Now we sought to identify signaling molecules necessary for T3 stimulation of Na,K-ATPase activity in alveolar epithelial cells. Whereas protein kinase A inhibitor H-8 and protein kinase C inhibitor bisindolymaleimide did not block the T3-induced increase in Na,K-ATPase activity, two inhibitors of phosphoinositide 3-kinase (PI3K), wortmannin and Ly294002, and two Src kinase inhibitors, PP1 and PP2, blocked the T3-induced Na,K ATPase activity. T3 stimulated the activity of PI3K as measured by phosphatidylinositol 3-phosphate. T3 also stimulated the serine 473 phosphorylation of the PI3K downstream molecule PKB/Akt in a dose-dependent manner. Transient expression of a constitutively active mutant of the PI3K catalytic subunit p110 augmented Na,K-ATPase activity and increased the amount of cell surface Na,K-ATPase alpha(1) subunit protein. T3 also stimulated Src family kinase activity. Transient expression of a constitutively active Src kinase increased Na,K-ATPase activity, PI3K activity, and phosphorylation of PKB/Akt at serine 473. PP1 or PP2 blocked T3-stimulated PKB/Akt phosphorylation at serine 473 and PI3K activity that was activated by an active mutant of Src; however, wortmannin did not inhibit the T3-stimulated Src kinase activity. Although PP1 and wortmannin abolished the increase in Na,K-ATPase activity induced by the active mutant of Src, PP1 did not inhibit the active mutant of PI3K-up-regulated Na,K-ATPase activity. In summary, T3 stimulates the PI3K/PKB pathway via the Src family of tyrosine kinases, and activation of both the Src family kinases and PI3K is required for the T3-induced stimulation of Na,K-ATPase activity and its cell surface expression in adult rat alveolar epithelial cells. PMID- 15342624 TI - Evolution of the biphenyl dioxygenase BphA from Burkholderia xenovorans LB400 by random mutagenesis of multiple sites in region III. AB - It is now established that several amino acids of region III of the biphenyl dioxygenase (BPDO) alpha subunit are involved in substrate recognition and regiospecificity toward chlorobiphenyls. However, the sequence pattern of the amino acids of that segment of seven amino acids located in the C-terminal portion of the alpha subunit is rather limited in BPDOs of natural occurrence. In this work, we have randomly mutated simultaneously four residues (Thr(335) Phe(336)-Ile(338)-Ile(341)) of region III of Burkholderia xenovorans LB400 BphA. The library was screened for variants able to oxygenate 2,2'-dichlorobiphenyl (2,2'-CB). Replacement of Phe(336) with Met or Ile with a concomitant change of Thr(335) to Ala created new variants that transformed 2,2'-CB into 3,4-dihydro 3,4-dihydroxy-2,2'-dichlorobiphenyl, which is a dead end metabolite that was not cleaved by BphC. Replacement of Thr(335)-Phe(336) with Ala(335)-Leu(336) did not cause this type of phenotypic change. Regiospecificity toward congeners other than 2,2'-CB that were oxygenated more efficiently by variant Ala(335)-Met(336) than by LB400 BPDO was similar for both enzymes. Thus structural changes that altered the regiospecificity toward 2,2'-CB did not affect the metabolite profile of other congeners, although it affected the rate of conversion of these congeners. It was especially noteworthy that both LB400 BPDO and the Ala(335) Met(336) variant generated 2,3-dihydroxy-2',4,4'-trichlorobiphenyl as the sole metabolite from 2,4,2',4'-CB and 4,5-dihydro-4,5-dihydroxy-2,3,2',3' tetrachlorobiphenyl as the major metabolite from 2,3,2',3'-CB. This shows that 2,4,2',4'-CB is oxygenated principally onto vicinal ortho-meta carbons 2 and 3 and that 2,3,2',3'-CB is oxygenated onto meta-para carbons 4 and 5 by both enzymes. The data suggest that interactions between the chlorine substitutes on the phenyl ring and specific amino acid residues of the protein influence the orientation of the phenyl ring inside the catalytic pocket. PMID- 15342625 TI - Revisiting the regiospecificity of Burkholderia xenovorans LB400 biphenyl dioxygenase toward 2,2'-dichlorobiphenyl and 2,3,2',3'-tetrachlorobiphenyl. AB - 2,2'-Dichlorobiphenyl (CB) is transformed by the biphenyl dioxygenase of Burkholderia xenovorans LB400 (LB400 BPDO) into two metabolites (1 and 2). The most abundant metabolite, 1, was previously identified as 2,3-dihydroxy-2' chlorobiphenyl and was presumed to originate from the initial attack by the oxygenase on the chlorine-bearing ortho carbon and on its adjacent meta carbon of one phenyl ring. 2,3,2',3'-Tetrachlorobiphenyl is transformed by LB400 BPDO into two metabolites that had never been fully characterized structurally. We determined the precise identity of the metabolites produced by LB400 BPDO from 2,2'-CB and 2,3,2',3'-CB, thus providing new insights on the mechanism by which 2,2'-CB is dehalogenated to generate 2,3-dihydroxy-2'-chlorobiphenyl. We reacted 2,2'-CB with the BPDO variant p4, which produces a larger proportion of metabolite 2. The structure of this compound was determined as cis-3,4-dihydro 3,4-dihydroxy-2,2'-dichlorobiphenyl by NMR. Metabolite 1 obtained from 2,2'-CB d(8) was determined to be a dihydroxychlorobiphenyl-d(7) by gas chromatographic mass spectrometric analysis, and the observed loss of only one deuterium clearly shows that the oxygenase attack occurs on carbons 2 and 3. An alternative attack at the 5 and 6 carbons followed by a rearrangement leading to the loss of the ortho chlorine would have caused the loss of more than one deuterium. The major metabolite produced from catalytic oxygenation of 2,3,2',3'-CB by LB400 BPDO was identified by NMR as cis-4,5-dihydro-4,5-dihydroxy-2,3,2',3'-tetrachlorobiphenyl. These findings show that LB400 BPDO oxygenates 2,2'-CB principally on carbons 2 and 3 and that BPDO regiospecificity toward 2,2'-CB and 2,3,2,',3'-CB disfavors the dioxygenation of the chlorine-free ortho-meta carbons 5 and 6 for both congeners. PMID- 15342626 TI - Irreversible thermal denaturation of glucose oxidase from Aspergillus niger is the transition to the denatured state with residual structure. AB - Glucose oxidase (GOX; beta-d-glucose:oxygen oxidoreductase) from Aspergillus niger is a dimeric flavoprotein with a molecular mass of 80 kDa/monomer. Thermal denaturation of glucose oxidase has been studied by absorbance, circular dichroism spectroscopy, viscosimetry, and differential scanning calorimetry. Thermal transition of this homodimeric enzyme is irreversible and, surprisingly, independent of GOX concentration (0.2-5.1 mg/ml). It has an apparent transition temperature of 55.8 +/- 1.2 degrees C and an activation energy of approximately 280 kJ/mol, calculated from the Lumry-Eyring model. The thermally denatured state of GOX after recooling has the following characteristics. (i) It retains approximately 70% of the native secondary structure ellipticity; (ii) it has a relatively low intrinsic viscosity, 7.5 ml/g; (iii) it binds ANS; (iv) it has a low Stern-Volmer constant of tryptophan quenching; and (v) it forms defined oligomeric (dimers, trimers, tetramers) structures. It is significantly different from chemically denatured (6.67 m GdmHCl) GOX. Both the thermal and the chemical denaturation of GOX cause dissociation of the flavin cofactor; however, only the chemical denaturation is accompanied by dissociation of the homodimeric GOX into monomers. The transition temperature is independent of the protein concentration, and the properties of the thermally denatured protein indicate that thermally denatured GOX is a compact structure, a form of molten globule-like apoenzyme. GOX is thus an exceptional example of a relatively unstable mesophilic dimeric enzyme with residual structure in its thermally denatured state. PMID- 15342627 TI - The auxiliary protein HypX provides oxygen tolerance to the soluble [NiFe] hydrogenase of ralstonia eutropha H16 by way of a cyanide ligand to nickel. AB - The hypX gene of the facultative lithoautotrophic bacterium Ralstonia eutropha is part of a cassette of accessory genes (the hyp cluster) required for the proper assembly of the active site of the [NiFe]-hydrogenases in the bacterium. A deletion of the hypX gene led to a severe growth retardation under lithoautotrophic conditions with 5 or 15% oxygen, when the growth was dependent on the activity of the soluble NAD+ -reducing hydrogenase. The enzymatic and infrared spectral properties of the soluble hydrogenase purified from a HypX negative strain were compared with those from an enzyme purified from a HypX positive strain. In activity assays under anaerobic conditions both enzyme preparations behaved the same. Under aerobic conditions, however, the mutant enzyme became irreversibly inactivated during H2 oxidation with NAD+ or benzyl viologen as the electron acceptor. Infrared spectra and chemical determination of cyanide showed that one of the four cyanide groups in the wild-type enzyme was missing in the mutant enzyme. The data are consistent with the proposal that the HypX protein is specifically involved in the biosynthetic pathway that delivers the nickel-bound cyanide. The data support the proposal that this cyanide is crucial for the enzyme to function under aerobic conditions. PMID- 15342628 TI - Intrinsic promoter recognition by a "core" RNA polymerase. AB - Two classes of RNA polymerases transcribe RNA from promoters on DNA templates: promoter recognition-competent single polypeptides and multisubunit enzymes that require separable promoter recognition factors. Eukaryotic mitochondria utilize an unusual hybrid of these classes composed of a "core" RNA polymerase related to the single polypeptide enzymes plus a "specificity factor" necessary for promoter utilization. Using supercoiled or premelted templates, we have discovered that the yeast core mitochondrial RNA polymerase (Rpo41) has the intrinsic ability to initiate from promoters without its specificity factor (Mtf1). Rpo41 requires the mitochondrial promoter sequence (ATATAAGTA) for this activity. On premelted templates addition of Mtf1 actually inhibits the promoter selective activity of Rpo41. Mtf1 increases abortive relative to productive transcription by Rpo41, possibly by stabilizing the promoter complex and reducing escape into elongation. The requirement for Mtf1 on closed but not open templates indicates that Mtf1 facilitates melting but not recognition of promoters. PMID- 15342629 TI - Coupling DNA supercoiling to transcription in defined protein systems. AB - Transcription of closed circular DNA templates in the presence of DNA gyrase is known to stimulate negative DNA supercoiling both in vivo and in vitro. It has proven elusive, however, to establish a general system in vitro that supports transcription-coupled DNA supercoiling (TCDS) by the "twin-domain" mechanism (Liu, L. F. and Wang, J. C. (1987) Proc. Natl. Acad. Sci. USA 84, 7024-7027) that operates in bacteria. In this report, we examine the properties of TCDS in defined protein systems that minimally contained T7 RNA polymerase and DNA gyrase. Specifically designed plasmid DNA templates permitted us to control the location and length of RNA transcripts. We demonstrate that TCDS takes place by two separate, and apparently independent, mechanistic pathways in vitro. The first supercoiling pathway, which is not likely to be significant in vivo, was found to be dependent on R-loop formation and could be suppressed by the presence of RNase H or bacterial HU protein. The second pathway for TCDS was much more potent, but became predominant in vitro only when sequence-specific DNA-bending proteins were present during transcription, and RNA transcript lengths exceeded 3 kb. This major supercoiling route was shown to be resistant to RNase H and had functional properties consistent with those predicted for the twin-domain mechanism. For example, DNA supercoiling activity was proportional to RNA transcript length and was greatly stimulated by macromolecular crowding agents. Under optimal conditions, the twin domain pathway of TCDS rapidly and efficiently generated superhelicity levels more than twice that typically found in vivo. PMID- 15342630 TI - Processing and joining of DNA ends coordinated by interactions among Dnl4/Lif1, Pol4, and FEN-1. AB - The repair of DNA double-strand breaks is critical for maintaining genetic stability. In the non-homologous end-joining pathway, DNA ends are brought together by end-bridging factors. However, most in vivo DNA double-strand breaks have terminal structures that cannot be directly ligated. Thus, the DNA ends are aligned using short regions of sequence microhomology followed by processing of the aligned DNA ends by DNA polymerases and nucleases to generate ligatable termini. Genetic studies in Saccharomyces cerevisiae have implicated the DNA polymerase Pol4 and the DNA structure-specific endonuclease FEN-1(Rad27) in the processing of DNA ends to be joined by Dnl4/Lif1. In this study, we demonstrated that FEN-1(Rad27) physically and functionally interacted with both Pol4 and Dnl4/Lif1 and that together these proteins coordinately processed and joined DNA molecules with incompatible 5' ends. Because Pol4 also interacts with Dnl4/Lif1, our results have revealed a series of pair-wise interactions among the factors that complete the repair of DNA double-strand breaks by non-homologous end joining and provide a conceptual framework for delineating the end-processing reactions in higher eukaryotes. PMID- 15342631 TI - UV light-induced DNA damage and tolerance for the survival of nucleotide excision repair-deficient human cells. AB - DNA damage can cause cell death unless it is either repaired or tolerated. The precise contributions of repair and tolerance mechanisms to cell survival have not been previously evaluated. Here we have analyzed the cell killing effect of the two major UV light-induced DNA lesions, cyclobutane pyrimidine dimers (CPDs) and 6-4 pyrimidine-pyrimidone photoproducts (6-4PPs), in nucleotide excision repair-deficient human cells by expressing photolyase(s) for light-dependent photorepair of either or both lesions. Immediate repair of the less abundant 6 4PPs enhances the survival rate to a similar extent as the immediate repair of CPDs, indicating that a single 6-4PP lesion is severalfold more toxic than a CPD in the cells. Because UV light-induced DNA damage is not repaired at all in nucleotide excision repair-deficient cells, proliferation of these cells after UV light irradiation must be achieved by tolerance of the damage at replication. We found that RNA interference designed to suppress polymerase zeta activity made the cells more sensitive to UV light. This increase in sensitivity was prevented by photorepair of 6-4PPs but not by photorepair of CPDs, indicating that polymerase zeta is involved in the tolerance of 6-4PPs in human cells. PMID- 15342632 TI - Proliferating cell nuclear antigen-dependent coordination of the biological functions of human DNA polymerase iota. AB - Y-family DNA polymerases are believed to facilitate the replicative bypass of damaged DNA in a process commonly referred to as translesion synthesis. With the exception of DNA polymerase eta (poleta), which is defective in humans with the Xeroderma pigmentosum variant (XP-V) phenotype, little is known about the cellular function(s) of the remaining human Y-family DNA polymerases. We report here that an interaction between human DNA polymerase iota (poliota) and the proliferating cell nuclear antigen (PCNA) stimulates the processivity of poliota in a template-dependent manner in vitro. Mutations in one of the putative PCNA binding motifs (PIP box) of poliota or the interdomain connector loop of PCNA diminish the binding between poliota and PCNA and concomitantly reduce PCNA dependent stimulation of poliota activity. Furthermore, although retaining its capacity to interact with poleta in vivo, the poliota-PIP box mutant fails to accumulate in replication foci. Thus, PCNA, acting as both a scaffold and a modulator of the different activities involved in replication, appears to recruit and coordinate replicative and translesion DNA synthesis polymerases to ensure genome integrity. PMID- 15342633 TI - Mechanistic analysis of pause site-dependent and -independent recombinogenic strand transfer from structurally diverse regions of the HIV genome. AB - Retroviral recombinants are generated by strand transfers occurring within internal regions of the viral genome and are a major source of genetic variability. Strand transfer has been linked to "pausing" occurring at secondary structures during synthesis by reverse transcriptase. Yet, weakly structured templates lacking strong pause sites also undergo efficient transfer. In this report, transfer crossover sites on high and low structured templates from the gag-pol frameshift region (GagPol) and the env (Env) regions, respectively, were determined by using a reconstituted in vitro strand transfer assay. The assay tested transfers occurring between a donor and acceptor template over a 150 nucleotide homologous region. The majority of crossovers were in a small 23 nucleotide region near a major pause site on GagPol, clearly indicating a pause driven mechanism. In contrast, on Env, transfers were more dispersed clustering toward the end of the homologous region. Slowing down polymerization on Env by decreasing the dNTP concentration resulted in crossovers shifting toward the beginning of the homologous region. Removal of a small 38-nucleotide region at the 3'-end of the Env acceptor had a large effect on the level of strand transfer despite very few crossovers mapping to this region. This implicated this part of the acceptor in transfers occurring at downstream positions. For Env the results support a mechanism where the acceptor rapidly binds nascent DNA, then "zippers" downstream catching up with the donor-DNA hybrid and displacing the donor. Such a mechanism may be important to recombination in low structure regions of the HIV genome. PMID- 15342634 TI - The ISG15 isopeptidase UBP43 is regulated by proteolysis via the SCFSkp2 ubiquitin ligase. AB - The Skp2 oncoprotein belongs to the family of F-box proteins that function as substrate recognition factors for SCF (Skp1, cullin, F-box protein) E3 ubiquitin ligase complexes. Binding of the substrate to the SCFSkp2 complex catalyzes the conjugation of ubiquitin molecules to the bound substrate, resulting in multi ubiquitination and rapid degradation by the 26 S proteasome. Using Skp2 as bait in a yeast two-hybrid screen, we have identified UBP43 as a novel substrate for Skp2. UBP43 belongs to the family of ubiquitin isopeptidases and specifically cleaves ISG15, a ubiquitin-like molecule that is induced by cellular stresses, such as type 1 interferons (IFN), nephrotoxic damage, and bacterial infection. UBP43 was originally identified as an up-regulated gene in knock-in mice expressing an acute myelogenous leukemia fusion protein, AML1-ETO, as well as in melanoma cell lines treated with IFN-beta. The phenotype of UBP43 knockout mice includes shortened life span, hypersensitivity to IFN, and neuronal damage, suggesting that tight regulation of ISG15 conjugation is critical for normal cellular function. In this study, we demonstrate that UBP43 is ubiquitinated in vivo and accumulates in cells treated with proteasome inhibitors. We also show that Skp2 promotes UBP43 ubiquitination and degradation, resulting in higher levels of ISG15 conjugates. In Skp2-/- mouse cells, levels of UBP43 are consistently up-regulated, whereas levels of ISG15 conjugates are reduced. Our results demonstrate that the SCFSkp2 is involved in controlling UBP43 protein levels and may therefore play an important role in modulating type 1 IFN signaling. PMID- 15342635 TI - Protein kinase CK2 is an inhibitor of the neuronal Cdk5 kinase. AB - The complex of Cdk5 and its neuronal activator p35 is a proline-directed Ser/Thr kinase that plays an important role in various neuronal functions. Deregulation of the Cdk5 enzymatic activity was found to associate with a number of neurodegenerative diseases. To search for regulatory factors of Cdk5-p35 in the brain, we developed biochemical affinity isolation using a recombinant protein comprising the N-terminal 149 amino acids of p35. The catalytic alpha-subunit of protein kinase CK2 (formerly known as casein kinase 2) was identified by mass spectrometry from the isolation. The association of CK2 with p35 and Cdk5 was demonstrated, and the CK2-binding sites were delineated in p35. Furthermore, CK2 displayed strong inhibition toward the Cdk5 activation by p35. The Cdk5 inhibition is dissociated from the kinase function of CK2 because the kinase-dead mutant of CK2 displayed the similar Cdk5 inhibitory activity as the wild-type enzyme. Further characterization showed that CK2 blocks the complex formation of Cdk5 and p35. Together, these findings suggest that CK2 acts as an inhibitor of Cdk5 in the brain. PMID- 15342636 TI - Cytoplasmic targeting signals in transmembrane invariant surface glycoproteins of trypanosomes. AB - Protein targeting mechanisms in flagellated protozoan parasites have received considerable interest because of a huge bias in these organisms toward the glycosylphosphatidylinositol anchor as a mechanism for the membrane attachment of cell surface macromolecules. In this study, the trafficking of invariant surface glycoprotein 65 (ISG65), a family of type I transmembrane proteins, was examined. Analysis of the C-terminal domains of ISG65 family members demonstrated a high level of conservation and, in particular, the presence of three lysine residues contained within the cytoplasmic tails of all ISG65s. ISG65 was expressed on the cell surface, in agreement with earlier work, but an intracellular pool of ISG65 was also detected within a Rab5A early endosome. Transplantation of the C terminal 74 amino acids of ISG65 (encompassing the 23 C-terminal residues of the extracellular domain, the transmembrane peptide, and the cytoplasmic domain) onto the N-terminal domain of BiP (BiPN) was sufficient to target the chimera to the same internal compartments as native ISG65. Further, site-directed mutagenesis indicated that the cytoplasmic tail was required for endoplasmic reticulum exit and that at least two of the cytoplasmic domain lysine residues are needed for endosomal targeting, as removal of all three led to surface expression. Kinetic measurements demonstrate that the BiPN fusion protein (containing the ISG65 C terminus) has a short half-life, indicating rapid turnover. In contrast, BiPN fusion proteins containing a glycosylphosphatidylinositol anchor instead of the ISG65 C-terminal region are stably expressed on the surface, confirming the requirement for the ISG65 sequence for endosomal targeting. We suggest that the lack of surface expression of the BiPN-ISG65 fusion protein is likely due to more efficient internalization compared with ISG65. Taken together, these data demonstrate the presence of a lysine-dependent endocytosis signal in the ISG65 family. PMID- 15342637 TI - Human but not murine toll-like receptor 2 discriminates between tri-palmitoylated and tri-lauroylated peptides. AB - Toll-like receptors (TLRs) mediate activation of the immune system upon challenge with microbial agonists, components of disintegrating cells of the body, or metabolic intermediates of lipidic nature. Comparison of murine (m) and human (h) TLR2 primary sequences revealed 65% of identical residues within the extracellular domains in contrast to 84% in the intracellular domains. Comparative analysis of TLR2-driven cell activation by various TLR2 agonists showed that the tri-lauroylated lipopeptide analog (Lau(3)CSK(4)) is recognized efficiently through mTLR2 but not hTLR2. Genetically complemented human embryonic kidney 293 cells and murine TLR2(-/-) embryonic fibroblasts, as well as human and murine macrophage cells, were used for this analysis. In contrast to cellular activation, which depended on blockable access of the TLR2-ligand to TLR2, cellular uptake of Lau(3)CSK(4) and tri-palmitoylated peptide (P(3)CSK(4)) was independent of TLR2. A low-conserved region spanning from leucine-rich repeat (LRR) motif 7 to 10 was found to control TLR2 species-specific cell activation. Exchange of mLRR8 for hLRR8 in mTLR2 abrogated mTLR2-typical cell activation upon cellular challenge with Lau(3)CSK(4) but not P(3)CSK(4), implicating mLRR8 as a central element of Lau(3)CSK(4) recognition. The point mutation L112P within LRR3 abrogated hTLR2-dependent recognition of lipopeptides but merely attenuated mTLR2 function, whereas deletion of the N-terminal third of each LRR-rich domain (LRRs 1 to 7) had the opposite effect on P(3)CSK(4) recognition. Despite similar domain structure of both TLR2 molecules, species-specific properties thus exist. Our results imply distinct susceptibilities of humans and mice to challenge with specific TLR2 ligands. PMID- 15342638 TI - Angiotensin receptor type 1 forms a complex with the transient outward potassium channel Kv4.3 and regulates its gating properties and intracellular localization. AB - We report a novel signal transduction complex of the angiotensin receptor type 1. In this complex the angiotensin receptor type 1 associates with the potassium channel alpha-subunit Kv4.3 and regulates its intracellular distribution and gating properties. Co-localization of Kv4.3 with angiotensin receptor type 1 and fluorescent resonance energy transfer between those two proteins labeled with cyan and yellow-green variants of green fluorescent protein revealed that Kv4.3 and angiotensin receptor type I are located in close proximity to each other in the cell. The angiotensin receptor type 1 also co-immunoprecipitates with Kv4.3 from canine ventricle or when co-expressed with Kv4.3 and its beta-subunit KChIP2 in human embryonic kidney 293 cells. Treatment of the cells with angiotensin II results in the internalization of Kv4.3 in a complex with the angiotensin receptor type 1. When stimulated with angiotensin II, angiotensin receptors type 1 modulate gating properties of the remaining Kv4.3 channels on the cell surface by shifting their activation voltage threshold to more positive values. We hypothesize that the angiotensin receptor type 1 provides its internalization molecular scaffold to Kv4.3 and in this way regulates the cell surface representation of the ion channel. PMID- 15342639 TI - The Traf2- and Nck-interacting kinase as a putative effector of Rap2 to regulate actin cytoskeleton. AB - Rap2 belongs to the Ras family of small GTP-binding proteins, but its specific roles in cell signaling remain unknown. In the present study, we have affinity purified from rat brain a Rap2-interacting protein of approximately 155 kDa, p155. By liquid chromatography tandem mass spectrometry, we have identified p155 as Traf2- and Nck-interacting kinase (TNIK). TNIK possesses an N-terminal kinase domain homologous to STE20, the Saccharomyces cerevisiae mitogen-activated protein kinase kinase kinase kinase, and a C-terminal regulatory domain termed the citron homology (CNH) domain. TNIK induces disruption of F-actin structure, thereby inhibiting cell spreading. In addition, TNIK specifically activates the c Jun N-terminal kinase (JNK) pathway. Among our observations, TNIK interacted with Rap2 through its CNH domain but did not interact with Rap1 or Ras. TNIK interaction with Rap2 was dependent on the intact effector region and GTP-bound configuration of Rap2. When co-expressed in cultured cells, TNIK colocalized with Rap2, while a mutant TNIK lacking the CNH domain did not. Rap2 potently enhanced the inhibitory function of TNIK against cell spreading, but this was not observed for the mutant TNIK lacking the CNH domain. Rap2 did not significantly enhance TNIK-induced JNK activation, but promoted autophosphorylation and translocation of TNIK to the detergent-insoluble cytoskeletal fraction. These results suggest that TNIK is a specific effector of Rap2 to regulate actin cytoskeleton. PMID- 15342640 TI - The biochemical characterization of two carotenoid cleavage enzymes from Arabidopsis indicates that a carotenoid-derived compound inhibits lateral branching. AB - Enzymes that are able to oxidatively cleave carotenoids at specific positions have been identified in animals and plants. The first such enzyme to be identified was a nine-cis-epoxy carotenoid dioxygenase from maize, which catalyzes the rate-limiting step of abscisic acid biosynthesis. Similar enzymes are necessary for the synthesis of vitamin A in animals and other carotenoid derived molecules in plants. In the model plant, Arabidopsis, there are nine hypothetical proteins that share some degree of sequence similarity to the nine cis-epoxy carotenoid dioxygenases. Five of these proteins appear to be involved in abscisic acid biosynthesis. The remaining four proteins are expected to catalyze other carotenoid cleavage reactions and have been named carotenoid cleavage dioxygenases (CCDs). The hypothetical proteins, AtCCD7 and AtCCD8, are the most disparate members of this protein family in Arabidopsis. The max3 and max4 mutants in Arabidopsis result from lesions in AtCCD7 and AtCCD8. Both mutants display a dramatic increase in lateral branching and are believed to be impaired in the synthesis of an unidentified compound that inhibits axillary meristem development. To determine the biochemical function of AtCCD7, the protein was expressed in carotenoid-accumulating strains of Escherichia coli. The activity of AtCCD7 was also tested in vitro with several of the most common plant carotenoids. It was shown that the recombinant AtCCD7 protein catalyzes a specific 9-10 cleavage of beta-carotene to produce the 10 black triangle down-apo beta-carotenal (C27) and beta-ionone (C13). When AtCCD7 and AtCCD8 were co expressed in a beta-carotene-producing strain of E. coli, the 13-apo-beta carotenone (C18) was produced. The C18 product appears to result from a secondary cleavage of the AtCCD7-derived C27 product. The sequential cleavages of beta carotene by AtCCD7 and AtCCD8 are likely the initial steps in the synthesis of a carotenoid-derived signaling molecule that is necessary for the regulation lateral branching. PMID- 15342641 TI - The crystal structure of an oxidatively stable subtilisin-like alkaline serine protease, KP-43, with a C-terminal beta-barrel domain. AB - The crystal structure of an oxidatively stable subtilisin-like alkaline serine protease, KP-43 from Bacillus sp. KSM-KP43, with a C-terminal extension domain, was determined by the multiple isomorphous replacements method with anomalous scattering. The native form was refined to a crystallographic R factor of 0.134 (Rfree of 0.169) at 1.30-A resolution. KP-43 consists of two domains, a subtilisin-like alpha/beta domain and a C-terminal jelly roll beta-barrel domain. The topological architecture of the molecule is similar to that of kexin and furin, which belong to the subtilisin-like proprotein convertases, whereas the amino acid sequence and the binding orientation of the C-terminal beta-barrel domain both differ in each case. Since the C-terminal domains of subtilisin-like proprotein convertases are essential for folding themselves, the domain of KP-43 is also thought to play such a role. KP-43 is known to be an oxidation-resistant protease among the general subtilisin-like proteases. To investigate how KP-43 resists oxidizing reagents, the structure of oxidized KP-43 was also determined and refined to a crystallographic R factor of 0.142 (Rfree of 0.212) at 1.73-A resolution. The structure analysis revealed that Met-256, adjacent to catalytic Ser-255, was oxidized similarly to an equivalent residue in subtilisin BPN'. Although KP-43, as well as proteinase K and subtilisin Carlsberg, lose their hydrolyzing activity against synthetic peptides after oxidation treatment, all of them retain 70-80% activity against proteinaceous substrates. These results, as well as the beta-casein digestion pattern analysis, have indicated that the oxidation of the methionine adjacent to the catalytic serine is not a dominant modification but might alter the substrate specificities. PMID- 15342642 TI - A GABAA receptor mutation linked to human epilepsy (gamma2R43Q) impairs cell surface expression of alphabetagamma receptors. AB - A mutation in the gamma2 subunit of the gamma-aminobutyric acid (GABA) type A receptor (GABAR), which changes an arginine to a glutamine at position 43 (R43Q), is linked to familial idiopathic epilepsies. We used radioligand binding, immunoblotting, and immunofluorescence techniques to examine the properties of wild-type alpha1beta2gamma2 and mutant alpha1beta2gamma2R43Q GABARs expressed in HEK 293 cells. The gamma2R43Q mutation had no affect on the binding affinity of the benzodiazepine flunitrazepam. However, in cells expressing alpha1beta2gamma2R43Q GABARs, the number of binding sites for [3H]flunitrazepam relative to wild-type receptors was decreased 75%. Using surface protein biotinylation, affinity purification, and immunoblotting, we demonstrated that expression of cell surface alpha1beta2gamma2R43Q GABARs was decreased. Surface immunostaining of HEK 293 cells expressing alpha1beta2gamma2R43Q GABARs confirmed that surface expression of the gamma2R43Q subunit was reduced. These data demonstrate that the gamma2R43Q mutation impairs expression of cell surface GABARs. A deficit in surface GABAR expression would reduce synaptic inhibition and result in neuronal hyperexcitability, which could explain why families possessing the gamma2R43Q subunit have epilepsy. PMID- 15342643 TI - The tryptophan-rich motifs of the thrombospondin type 1 repeats bind VLAL motifs in the latent transforming growth factor-beta complex. AB - Transforming growth factor-beta (TGF-beta) is secreted as a latent complex of the latency-associated peptide (LAP) and the mature domain, which must be activated for TGF-beta to signal. We previously identified thrombospondin 1 (TSP1) as a physiologic activator of TGF-beta in vitro and in vivo. The WSXW sequences in the type 1 repeats of TSP1 interact with the mature domain of TGF-beta, and WSXW peptides inhibit TSP1-mediated activation by blocking TSP1 binding to the TGF beta latent complex. However, the binding site for the WSXW sequence was not identified. In this report, we show that the WSXW sequences bind the (61)VLAL sequence in mature TGF-beta and also bind (77)VLAL in LAP. A glutathione S transferase (GST) fusion protein of the second TSP1 type 1 repeat (GST-TSR2) binds immobilized VLAL peptide. VLAL peptides inhibit binding of LAP and mature TGF-beta to soluble GST-TSR2 and immobilized WSXW peptide. VLAL peptide inhibits TSP1-mediated activation of recombinant and endothelial cell-derived latent TGF beta. Furthermore, TGF-beta or LAP deleted in the VLAL sequence fails to bind immobilized WSXW or soluble GST-TSR2, indicating that binding to both VLAL sequences is important for association of TSP1 and the latent complex. Additionally, TSP1 is unable to activate latent TGF-beta when VLAL is deleted from the mature domain. These data show that the WSXW motif binds VLAL on both LAP and mature TGF-beta, and these interactions are critical for TSP1-mediated activation of the TGF-beta latent complex. PMID- 15342644 TI - "Respirasome"-like supercomplexes in green leaf mitochondria of spinach. AB - Higher plant mitochondria have many unique features compared with their animal and fungal counterparts. This is to a large extent related to the close functional interdependence of mitochondria and chloroplasts, in which the two ATP generating processes of oxidative phosphorylation and photosynthesis, respectively, take place. We show that digitonin treatment of mitochondria contaminated with chloroplasts from spinach (Spinacia oleracea) green leaves at two different buffer conditions, performed to solubilize oxidative phosphorylation supercomplexes, selectively extracts the mitochondrial membrane protein complexes and only low amounts of stroma thylakoid membrane proteins. By analysis of digitonin extracts from partially purified mitochondria of green leaves from spinach using blue and colorless native electrophoresis, we demonstrate for the first time that in green plant tissue a substantial proportion of the respiratory complex IV is assembled with complexes I and III into "respirasome"-like supercomplexes, previously observed in mammalian, fungal, and non-green plant mitochondria only. Thus, fundamental features of the supramolecular organization of the standard respiratory complexes I, III, and IV as a respirasome are conserved in all higher eukaryotes. Because the plant respiratory chain is highly branched possessing additional alternative enzymes, the functional implications of the occurrence of respiratory supercomplexes in plant mitochondria are discussed. PMID- 15342645 TI - An inhibitor of O-glycosylation induces apoptosis in NIH3T3 cells and developing mouse embryonic mandibular tissues. AB - The family of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases (ppGaNTases) is responsible for initiating mucin-type O-linked glycosylation in higher eukaryotes. To begin to examine the biological role of O-linked glycosylation, mammalian cells were treated with a small molecule inhibitor (designated 1-68A, Ref. 15) of ppGaNTase activity. NIH3T3 cells exposed to the inhibitor were shown to undergo a significant reduction in cell surface O glycosylation as detected by staining with jacalin and peanut agglutinin lectins after 30 min of treatment; no reduction in staining using antibodies to O-linked N-acetylglucosamine or the lectin concanavalin A was detected. Apoptosis was also observed in treated cells after 45 min of exposure, ostensibly following the O glycosylation reduction. Overexpression of several different ppGaNTase isoforms restored cell surface O-glycosylation and rescued inhibitor-induced apoptosis. Additionally, mouse embryonic mandibular organ cultures exposed to 1-68A developed abnormally, presumably because of epithelial and mesenchymal apoptosis that followed a reduction in jacalin and peanut agglutinin staining. Our studies suggest that mucin-type O-linked glycosylation may be required for normal development and that ppGaNTases may play a role in the regulation of apoptosis. PMID- 15342646 TI - Synaptic activity augments muscarinic acetylcholine receptor-stimulated inositol 1,4,5-trisphosphate production to facilitate Ca2+ release in hippocampal neurons. AB - Intracellular Ca2+ store release contributes to activity-dependent synaptic plasticity in the central nervous system by modulating the amplitude, propagation, and temporal dynamics of cytoplasmic Ca2+ changes. However, neuronal Ca2+ stores can be relatively insensitive to increases in the store-mobilizing messenger inositol 1,4,5-trisphosphate (IP3). Using a fluorescent biosensor we have visualized M1 muscarinic acetylcholine (mACh) receptor signaling in individual hippocampal neurons and observed increased IP3 production in the absence of concurrent Ca2+ store release. However, coincident glutamate-mediated synaptic activity elicited enhanced and oscillatory IP3 production that was dependent upon ongoing mACh receptor stimulation and S-alpha-amino-3-hydroxy-5 methyl-4-isoazolepropionic acid receptor activation of Ca2+ entry. Moreover, the enhanced levels of IP3 now mobilized Ca2+ from intracellular stores that were refractory to the activation of mACh receptors alone. We conclude that convergent ionotropic and metabotropic receptor inputs can facilitate Ca2+ signaling by enhancing IP3 production as well as augmenting release by Ca2+-induced Ca2+ release. PMID- 15342647 TI - Trafficking of exogenous peptides into proteasome-dependent major histocompatibility complex class I pathway following enterotoxin B subunit mediated delivery. AB - The B-subunit component of Escherichia coli heat-labile enterotoxin (EtxB), which binds to cell surface GM1 ganglioside receptors, was recently shown to be a highly effective vehicle for delivery of conjugated peptides into the major histocompatibility complex (MHC) class I pathway. In this study we have investigated the pathway of epitope delivery. The peptides used contained the epitope either located at the C terminus or with a C-terminal extension. Pretreatment of cells with cholesterol-disrupting agents blocked transport of EtxB conjugates to the Golgi/endoplasmic reticulum, but did not affect EtxB mediated MHC class I presentation. Under these conditions, EtxB conjugates entered EEA1-positive early endosomes where peptides were cleaved and translocated into the cytosol. Endosome acidification was required for epitope presentation. Purified 20 S immunoproteasomes were able to generate the epitope from peptides in vitro, but 26 S proteasomes were not. Only presentation from the C-terminal extended peptide was proteasome-dependent in cells, and this was found to be significantly slower than presentation from peptides with the epitope at the C terminus. These results implicate the proteasome in the generation of the correct C terminus of the epitope and are consistent with proteasome-independent N-terminal trimming. Epitope presentation was blocked in a TAP-deficient cell line, providing further evidence that conjugated peptides enter the cytosol as well as demonstrating a requirement for the peptide transporter. Our findings demonstrate the utility of EtxB-mediated peptide delivery for rapid and efficient loading of MHC class I epitopes in several different cell types. Conjugated peptides are released from early endosomes into the cytosol where they gain access to proteasomes and TAP in the "classical" pathway of class I presentation. PMID- 15342648 TI - Draper-mediated and phosphatidylserine-independent phagocytosis of apoptotic cells by Drosophila hemocytes/macrophages. AB - The mechanism of phagocytic elimination of dying cells in Drosophila is poorly understood. This study was undertaken to examine the recognition and engulfment of apoptotic cells by Drosophila hemocytes/macrophages in vitro and in vivo. In the in vitro analysis, l(2)mbn cells (a cell line established from larval hemocytes of a tumorous Drosophila mutant) were used as phagocytes. When l(2)mbn cells were treated with the molting hormone 20-hydroxyecdysone, the cells acquired the ability to phagocytose apoptotic S2 cells, another Drosophila cell line. S2 cells undergoing cycloheximide-induced apoptosis exposed phosphatidylserine on their surface, but their engulfment by l(2)mbn cells did not seem to be mediated by phosphatidylserine. The level of Croquemort, a candidate phagocytosis receptor of Drosophila hemocytes/macrophages, increased in l(2)mbn cells after treatment with 20-hydroxyecdysone, whereas that of Draper, another candidate phagocytosis receptor, remained unchanged. However, apoptotic cell phagocytosis was reduced when the expression of Draper, but not of Croquemort, was inhibited by RNA interference in hormone-treated l(2)mbn cells. We next examined whether Draper is responsible for the phagocytosis of apoptotic cells in vivo using an assay for engulfment based on assessing DNA degradation of apoptotic cells in dICAD mutant embryos (which only occurred after ingestion by the phagocytes). RNA interference-mediated decrease in the level of Draper in embryos of mutant flies was accompanied by a decrease in the number of cells containing fragmented DNA. Furthermore, histochemical analyses of dispersed embryonic cells revealed that the level of phagocytosis of apoptotic cells by hemocytes/macrophages was reduced when Draper expression was inhibited. These results indicate that Drosophila hemocytes/macrophages execute Draper-mediated phagocytosis to eliminate apoptotic cells. PMID- 15342649 TI - AMP-activated protein kinase-regulated phosphorylation and acetylation of importin alpha1: involvement in the nuclear import of RNA-binding protein HuR. AB - Nuclear import of HuR, a shuttling RNA-binding protein, is associated with reduced stability of its target mRNAs. Increased function of the AMP-activated protein kinase (AMPK), an enzyme involved in responding to metabolic stress, was recently shown to reduce the cytoplasmic levels of HuR. Here, we provide evidence that importin alpha1, an adaptor protein involved in nuclear import, contributes to the nuclear import of HuR through two AMPK-modulated mechanisms. First, AMPK triggered the acetylation of importin alpha1 on Lys(22), a process dependent on the acetylase activity of p300. Second, AMPK phosphorylated importin alpha1 on Ser(105). Accordingly, expression of importin alpha1 proteins bearing K22R or S105A mutations failed to mediate the nuclear import of HuR in intact cells. Our results point to importin alpha1 as a critical downstream target of AMPK and key mediator of AMPK-triggered HuR nuclear import. PMID- 15342650 TI - Components of the DNA methylation system of chromatin control are RNA-binding proteins. AB - The view that autosomal gene expression is controlled exclusively by protein trans-acting factors has been challenged recently by the identification of RNA molecules that regulate chromatin. In the majority of cases where RNA molecules are implicated in DNA control, the molecular mechanisms are unknown, in large part because the RNA.protein complexes are uncharacterized. Here, we identify a novel set of RNA-binding proteins that are well known for their function in chromatin regulation. The RNA-interacting proteins are components of the mammalian DNA methylation system. Genomic methylation controls chromatin in the context of transposon silencing, imprinting, and X chromosome dosage compensation. DNA methyltransferases (DNMTs) catalyze methylation of cytosines in CGs. The methyl-CGs are recognized by methyl-DNA-binding domain (MBD) proteins, which recruit histone deacetylases and chromatin remodeling proteins to effect silencing. We show that a subset of the DNMTs and MBD proteins can form RNA.protein complexes. We characterize the MBD protein RNA-binding activity and show that it is distinct from the methyl-CG-binding domain and mediates a high affinity interaction with RNA. The RNA and methyl-CG binding properties of the MBD proteins are mutually exclusive. We speculate that DNMTs and MBD proteins allow RNA molecules to participate in DNA methylation-mediated chromatin control. PMID- 15342652 TI - The dental curriculum at North American dental institutions in 2002-03: a survey of current structure, recent innovations, and planned changes. AB - This study examined the current format of curricula at North American dental schools, determined curriculum evaluation strategies, and identified recently implemented changes as well as planned future innovations. The academic affairs deans of sixty-four North American dental schools received an email survey in August 2002; a second, follow-up survey was sent to nonresponders in February 2003. Online responses were collected and analyzed using SurveyTracker software. The final response rate was 87 percent, with forty-eight U.S. schools and eight Canadian schools responding. Respondents were asked to select descriptive statements about the general organization of their curricula and the degree to which problem-based learning (PBL), case-reinforced learning (CRL), curricular integration, and community-based clinical treatment experiences were incorporated. They were also requested to identify strategies employed to evaluate the curriculum and to report recently completed and desired future curriculum modifications. In regard to desired future curriculum innovations, respondents identified why they were considering curriculum changes and identified resources needed to implement the planned changes. Sixty-six percent of those who responded defined their current curriculum organization as primarily discipline-based with a few interdisciplinary courses. Nearly 60 percent of schools reported that they used PBL and CRL in specific courses or for components of certain courses, but only 5 percent of the respondents indicated that all of their courses used PBL. Regarding integration of major sections of the curriculum, only 7 percent reported that their entire curriculum was organized around themes of interrelated topics. Sixty-four percent reported that their curriculum had required community-based clinical treatment experiences for students. The most frequent innovations in the past three years were increased use of computer and web-based learning (86 percent), creation of patient care experiences early in the curriculum (84 percent), enhancement of competency evaluation methods (84 percent), and curriculum decompression (79 percent). These items plus increased community-based care were the most frequently identified future curricular innovations. There were virtually no differences between the responses of Canadian and U.S. dental schools. The results of this study help to broadly characterize dental curricula at North American dental institutions and identify curriculum modifications anticipated by the academic dean respondents. PMID- 15342653 TI - Perspectives on affirmative action in academic dental institutions: the U.S. Supreme Court rulings in the University of Michigan cases. AB - In June 2003 the U.S. Supreme Court upheld the constitutionality of using race as a factor in higher education admissions decisions. This article considers the impact of the Supreme Court decisions on admissions procedures at selected academic dental institutions (ADI) and their parent institutions. We interviewed fifty-eight leaders considered to be individual stakeholders at seven ADI and their related parent institutions, state dental associations, and state legislatures using a common set of questions about the Supreme Court decisions. Educators from the ADI and their parent institutions were consistent in their responses that the rulings upheld affirmative action as necessary to achieve diversity. State organized dentistry officials did not appear to be as aware as others of the rulings, whereas legislators were mixed in their responses. Except for the University of Michigan undergraduate admissions procedures, it remains to be seen what the impact will be for other higher education institutions and for academic dental institutions. Although the rulings have provided guidelines for achieving diversity using race/ ethnicity as one of several factors, the rulings will possibly be challenged, thus requiring vigilance on the part of parent institutions and their ADI to ensure compliance with the spirit of the rulings and to avoid attack from opponents of affirmative action. PMID- 15342654 TI - The pathway to dentistry for minority students: from their perspective. AB - The small number of minorities in the field of dentistry is a serious concern. While the United States as a whole has become more diverse with minorities making up 25 percent of the total U.S. population, only a handful (14 percent) are currently practicing dentistry, and only 11 percent entering dental schools are underrepresented minorities. Pipeline, Profession, & Practice: Community-Based Dental Education is a national foundation-sponsored program designed to address this issue in dental education. To understand the reasons why dentistry attracts so few underrepresented minority (URM) students, we conducted focus groups and in depth interviews to determine the challenges facing minority students when they apply to and attend dental school. Ten focus groups were conducted with a total of ninety-two minority students (fourteen undergraduate students and seventy eight students currently enrolled in dental schools) at six universities in four geographic regions. In addition, four in-depth interviews were held with faculty advisors who teach, mentor, and recruit minority students. The major findings of the study are as follows: 1) early and frequent exposure to dentistry and dentists in practice is essential for minority students to consider this profession; 2) while many dental schools have earnestly tried to recruit minority applicants, most URM students find out about dental programs by a family member or friend and not as a result of an intentional recruiting effort; and 3) hearing directly from minority students could be a solid first step in understanding the dental school experience from a different vantage point. This study has important implications for the methods dental schools use to both recruit minority students and foster a learning environment that is sensitive to students from diverse backgrounds. PMID- 15342655 TI - Characteristics of dental school feeder institutions. AB - A major challenge faced by all dental schools is the need to attract highly qualified student applicants. The purpose of this study was to use 2002-03 AADSAS data to identify and characterize feeder colleges and universities that are the major source of applicants to U.S. dental schools. Feeder schools were defined as any institutions with five or more applicants, and minority-feeder schools as those with two or more minority applicants. Feeder schools were ranked by their total numbers of applicants (Category 1) and by their ratio of applicants to total undergraduate enrollment (Category 2). Feeder institutions were compared using total enrollment, degree status, geographic distribution, religious affiliation, numbers of minority applicants, and college admissions selectivity criteria. The top fifty Category 1 schools had an average enrollment of over 19,000 students and an average of sixty-seven applicants. The top fifty Category 2 schools had an average enrollment of approximately 8,500 students and an average of forty-nine applicants. Less than 1 percent of applicants from the top feeder institutions attended the nation's most competitive schools. California and Utah accounted for 28 percent of the total applicants from feeder institutions, followed by Florida (6.2 percent) and New York (5.7 percent). Seventeen of the top twenty-five Category 2 schools (68 percent) were affiliated with or had student bodies associated with a particular religion, with the Seventh-Day Adventist and Mormon institutions accounting for 544 applicants. The majority of all applicants from feeder institutions attended schools in the Southwest. The majority of black and Hispanic feeder institutions were in Florida, Tennessee, Louisiana, and Puerto Rico. Results suggest that factors such as school size, geographic location, religious affiliation, and admissions selectivity criteria of colleges and universities may have a direct impact on the dental applicant pool. PMID- 15342656 TI - Hypertension in a dental school patient population. AB - Patient records were retrospectively reviewed to investigate the incidence of hypertensive patients seen at a U.S. dental school. This research was conducted to create an awareness of the current problems in diagnosing and treating hypertensive patients in the dental environment. Nine hundred and seventy-six records of patients seen between January 1, 1999 and January 1, 2000 were reviewed. Five hundred records that met specific study criteria related to health history, medications used, recorded blood pressure, and other criteria were selected for the study. Factors examined included demographic data consisting of age, sex, and ethnicity; history of hypertension; Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure classification; control of hypertension; and medications used. The data demonstrated that 32 percent of the patients were hypertensive, 49 percent of whom were unaware of their high blood pressure prior to their dental visit. Nearly 9 percent of the hypertensive patients with elevated blood pressure had to be immediately sent for medical consult before they could receive dental treatment. The average blood pressure of the hypertensive patients was systolic 145.6 and diastolic 87.9 with a range of 110 to 240 systolic, 60 to 135 diastolic. Of the diagnosed patients, 41.9 percent were taking antihypertensive medication for their condition, and 13 percent were taking two or more medications. Nearly one third of a sample of 500 dental school clinic patients had high blood pressure in this retrospective study. This study demonstrates that it is crucial that dental providers take blood pressure readings for screening, monitoring of hypertensive patients, and appropriate dental care. PMID- 15342657 TI - Developing competency in research management, entrepreneurship, and technology transfer: a workshop course. AB - In July 1999, the National Institute of Dental and Craniofacial Research (NIDCR) convened a Blue Ribbon Panel that recommended management skills, entrepreneurship, and technology transfer should be included in dental education. The panel's recommendations were implemented in an NIDCR-funded pilot project, "Workshop Course to Promote and Develop Dental Products and Technologies." The workshop consisted of lectures presented by seven faculty members recruited from academia, government, and business, along with an analysis of a professor's invention and the barriers encountered in transforming the invention into a product. Evaluation consisted of a pre- and post-workshop survey. The workshop was presented to twenty-two participants on November 8 and 9, 2003 at the University of Connecticut School of Dental Medicine and, to refine the presentation further, will be tested at five additional dental schools (University of Pennsylvania, Harvard University, New York University, Nova Southeastern University, and University of Southern California). The results indicated that the workshop's courses would be helpful to the commercialization of inventions. In addition, dental students with experience in basic research expressed an interest in research of projects of use in dental practice. These findings suggest that pursuing research and an academic career might be more appealing if their research was product-oriented. PMID- 15342658 TI - A communication skills course for undergraduate dental students. AB - Sixty-seven third-year dental students in Dunedin, New Zealand, participated in a communication skills course, using simulated patients, case-based scenarios, videotaped interviews, and class roleplays. The course introduced active listening techniques, taking a medical history, and emotion-handling skills. This course was adapted from an existing course for medical students run by the Department of Psychological Medicine, Dunedin School of Medicine. The results of the student evaluation questionnaire (n = 59) indicated that students rated the course very highly. Retrospective ratings indicated that the students considered communication skills to be significantly more important as a component of their undergraduate training after completion of the course than prior to it. As might be expected, students whose ratings were higher after the course also reported that the course helped them to develop new communication skills and techniques; increased their interest in the subject and their self-confidence; rated the tutor as more effective and the course materials as more helpful; and considered the course to be significantly more stimulating than those students whose ratings of the importance of communication skills remained the same or decreased. PMID- 15342659 TI - Taxonomy for competency-based dental curricula. AB - The objective of this article is to propose a classification of dental competencies. Interest in dental competencies has grown consistently during the last three decades. However, the dental education literature suggests that the term "competency" is understood and used differently by dental schools around the world. The taxonomic classification of dental competencies we propose follows a systematic approach starting at the highest level of complexity, i.e., the professional profile the teaching institution envisions for its graduates, and following in a decreasing degree of complexity to competency function, task, step, movement, and moment. This taxonomy has proved to be useful for more than thirty years in the Dental School of the Peruvian University Cayetano Heredia. Graduates of this school are successful practitioners, teachers, and researchers in Peru and other countries. The classification proposed here should clarify terms, facilitate curriculum design and learning assessment, stimulate further discussion on the matter, and facilitate communication among the dental education establishment. PMID- 15342660 TI - Reliability and validity of a manual dexterity test to predict preclinical grades. AB - The University of California, San Francisco School of Dentistry wanted to determine if a predental school manual dexterity test predicts: 1) subsequent grades in preclinical restorative courses, and 2) faculty perceptions of satisfactory performance in these skills that would indicate the student is ready to advance to the clinic. The study population was comprised of all 244 applicants admitted to UCSF School of Dentistry's D.D.S. program from Classes of 2000 to 2002 and who matriculated into the program. The manual dexterity test (MDT) consisted of a two-hour block-carving test. Three preclinical faculty, three clinical faculty, and two basic science faculty graded the blocks. Even after instruction and calibration, faculty varied greatly in their grading (intra rater reliability kappa statistics ranging from 0.34 to 1.00). Two of three preclinical raters gave No Passes for the MDT in 9.8 percent of the incoming, first-year dental students. Of these twenty-three students, only four (17 percent) were in the lower 10 percent of their classes according to their five preclinical restorative laboratory courses after two years, and four (33 percent) were among the twelve students the three preclinical laboratory directors identified as laboratory cautions. The MDT did not significantly (p=0.342) predict students in the bottom 10 percent after five restorative preclinical laboratory courses, above and beyond current admissions criteria. Among current admissions criteria, PAT score was the only item at least moderately correlated with preclinical average percentile class rank (Spearman correlation = 0.34). In conclusion, the MDT did not appear to add information to the current admissions criteria. PMID- 15342661 TI - Implementing evidence-based practice in undergraduate teaching clinics: a systematic review and recommendations. AB - The objective of this project was to identify an effective methodology of approaching and implementing evidence-based principles in undergraduate teaching clinics to promote evidence-based dentistry in future clinical practice. A systematic review was undertaken to examine evidence-based clinical teaching and faculty continuing education. Research published from 1996 to 2002 was retrieved by searching several databases and the Internet, along with conducting hand searches and reviewing bibliographies maintained by faculty experts. Qualitative checklists for different types of studies were created to evaluate the literature. Relevant studies were selected if they met all four predetermined essential criteria and a minimum of two out of three desirable criteria. Systematic reviews were chosen if they met all five essential criteria. Data from selected articles were extracted, and study quality was assessed. We found that three systematic reviews and nine original research articles were deemed methodologically acceptable. Problem-based learning and evidence-based health care interventions increased student knowledge of medical topics and their ability to search, evaluate, and appraise medical literature. Dental students in a problem-based learning curriculum, emphasizing evidence-based practices, scored higher on the NDB I (National Dentistry Boards, Part I) than students in traditional curricula. While effective modules in implementing theoretical evidence-based principles exist, very few high-quality studies are available that examine these principles in dental undergraduate teaching clinics. No methodologically appropriate studies on the education of faculty in the implementation of evidence-based principles are available. Some studies promote promising theories and methodologies of teaching evidence-based care; based on these theories, a comprehensive model is proposed in this article. Considering the strength of evidence in the reviewed literature, we concluded that an evidence-based approach to clinical care is effective. PMID- 15342662 TI - Annual ADEA survey of dental school seniors: 2003 graduating class. PMID- 15342663 TI - Clinical trials for malignant lymphoma in Japan. AB - The results of the clinical trials by the Lymphoma Study Group of the Japan Clinical Oncology Group (JCOG-LSG) and those of the industry-supported trials mainly conducted by the members of JCOG-LSG are summarized. In the treatment of advanced aggressive non-Hodgkin's lymphoma (NHL), we investigated the efficacy of granulocyte colony-stimulating factor (G-CSF)-supported, dose-intensified strategies. Based on the results of a randomized phase II study (JCOG9505), we conducted a phase III study, JCOG9809, comparing CHOP and biweekly CHOP. However, JCOG9809 was terminated early based on the results of a planned interim analysis, because it was deemed highly unlikely that biweekly CHOP would be superior to standard CHOP. For aggressive ATL, a G-CSF-supported, dose-intensified, multi agent regimen (JCOG9303; LSG15) showed superior efficacy to our historical controls. To establish a new standard for ATL, we conducted a phase III study, JCOG9801, comparing LSG15 and biweekly CHOP. To develop new agents for lymphoid malignancies, we focused on irinotecan hydrochloride, interferon-alpha, cladribine and oral fludarabine. Among them, cladribine and oral fludarabine are promising for indolent B-cell malignancies. The Japanese phase I and II studies of rituximab, a chimeric anti-CD20 monoclonal antibody, in relapsed indolent and aggressive B-NHL showed high efficacy with minimal toxicities, which led us to conduct combination studies with chemotherapy for B-NHL. In addition, a phase I study of a radiolabeled anti-CD20 antibody (ibritumomab tiuxetan) was completed in 2003, and a phase II study for indolent B-NHL will be initiated. The multicenter trials by the JCOG-LSG and industry-supported new agent studies will contribute to further improvement in the treatment of malignant lymphoma. PMID- 15342664 TI - Evaluation of oxidative stress and nitric oxide levels in patients with oral cavity cancer. AB - OBJECTIVE: The aim of this study was to evaluate the magnitude of oxidative stress and levels of nitric oxide in patients with oral cavity cancer by analyzing the levels of lipid peroxidation products, antioxidants and nitric oxide products. METHODS: This prospective study was conducted on 15 patients with biopsy proven squamous cell cancer of the oral cavity with clinical stage III/IV and an equal number of age and sex matched healthy subjects. The levels of lipid peroxidation products, antioxidants and nitric oxide products were determined by colorimetric methods. RESULTS: Lipid peroxidation products like lipid hydroperoxide (LHP) and malondialdehyde (MDA) and nitric oxide products like nitrite (NO(2)(-)), nitrate (NO(3)(-)) and total nitrite (TNO(2)(-)) were significantly elevated, whereas enzymatic and non-enzymatic antioxidants were significantly lowered in oral cavity cancer patients when compared to normal healthy subjects. CONCLUSIONS: Enhanced lipid peroxidation with concomitant decrease in antioxidants is indicative of oxidative stress that provides evidence of the relationship between lipid peroxidation and oral cavity cancer. Increased nitric oxide production represents a general mechanism in its pathogenesis. PMID- 15342665 TI - Management of primary malignant germ cell tumor of the mediastinum. AB - BACKGROUND: Primary mediastinal malignant germ cell tumors (GCTs) are rare and have a worse prognosis than their gonadal counterparts. Although multimodality treatment is a standard therapeutic strategy in mediastinal GCTs, the clinical implications of surgical intervention remain unclear. METHODS: Forty-eight patients with primary mediastinal malignant GCT who were treated at the National Cancer Center Hospital, Tokyo, from 1962 to 2002 were studied retrospectively with regard to their histology and clinical profile. RESULTS: Mediastinal GCT occurred predominantly in young males, with a mean age of 28.8 years at the time of diagnosis. There were 46 males (96%) and two females (4%). Histologically, seven patients (15%) were diagnosed as having pure seminoma and 41 (85%) had non seminomatous GCT. Treatment consisted of surgery alone in nine patients, surgery followed by chemotherapy in two, and chemotherapy followed by surgery in 20. The other 17 patients received chemotherapy and/or radiotherapy without surgery. Of these latter 17 patients, 14 developed progressive disease and three were followed up with a sustained partial response. Among the 31 patients who underwent surgery, complete resection was performed in 27 (87%) and incomplete resection was performed in four (13%). Twelve (41%) patients had elevated serum tumor marker levels preoperatively. Among the 20 patients who received preoperative chemotherapy, viable cells were found in the resected specimen in six (30%). With regard to tumor recurrence in patients with surgical intervention, the preoperative serum tumor marker levels and the presence of viable cells in the resected specimen were significantly associated with recurrence. There was no significant association between surgical curability and recurrence. The 5-year overall survival rate in all 48 patients was 45.5%. CONCLUSIONS: Surgical intervention for mediastinal GCT may be needed to remove a chemotherapy-refractory tumor or to assess the pathological response to chemotherapy to determine the indications for further chemotherapy. PMID- 15342666 TI - Validation of the Functional Assessment of Cancer Therapy-General (FACT-G) scale for measuring the health-related quality of life in Korean women with breast cancer. AB - BACKGROUND: The Functional Assessment of Cancer Therapy-General (FACT-G) scale, which was developed and validated in the USA, is widely used to measure the health-related quality of life in cancer patients. The purpose of the present study was to empirically validate the FACT-G scale with Korean breast cancer patients. METHODS: A convenience sample of 193 women with breast cancer was recruited from a university hospital. The subjects were asked to complete the Korean version of the FACT-G scale. The data were analyzed using exploratory factor analysis with varimax rotation to determine factor construct validity. The loading criterion was set at 0.40 and above, inter-subscale correlations were computed using Pearson correlation, and the reliability of the internal consistency for the total scale and its subscales were assessed by Cronbach's alpha. RESULTS: The factor structure of the Korean version of the FACT-G scale paralleled that of the English version: the physical, social/family, emotional, and functional well-being subscales were constructively valid in Korean breast cancer patients. However, there is the possibility of culture-specific differences in the social/family well-being subscale, and some problematic translations were revealed. Cronbach's alpha for the total scale was 0.89 and that for the subscales ranged from 0.78 to 0.90. CONCLUSION: The Korean version of the FACT-G scale was demonstrated as reliable and valid. Therefore, the scale can be used in research and clinical settings to assess the quality of life of Korean breast cancer patients. PMID- 15342667 TI - Single-agent capecitabine in patients with metastatic colorectal cancer refractory to 5-fluorouracil/leucovorin chemotherapy. AB - OBJECTIVE: The effectiveness of capecitabine, an oral fluoropyrimidine carbamate, is well documented in previously untreated metastatic colorectal cancer patients (overall response rate: 25%). However, its efficacy in patients with metastatic colorectal cancer refractory to 5-fluorouracil/leucovorin (5-FU/LV) has not been determined. This study was performed to evaluate the efficacy and to identify the side-effects of capecitabine in patients with metastatic colorectal cancer showing progression despite 5-FU/LV-based combination chemotherapy. METHODS: Fifty-one metastatic colorectal cancer patients who showed progressive disease in 5-FU/LV-containing regimens (median: two regimes) were treated with capecitabine 1,250 mg/m(2) twice daily (days 1-14 repeated every 3 weeks). RESULTS: Only one partial response was observed (response rate: 2%). Twenty-seven patients (53%) showed stable disease after two cycles. The median time to disease progression of either a partial response or stable disease was 3.4 months. Hand-foot syndrome was the main toxicity of capecitabine and occurred in 35% of cases (grade 3 or 4 in 6%). The median number of cycles administered was two and the relative dose intensity of capecitabine was 80%. CONCLUSION: The response rate to capecitabine was low in metastatic colorectal cancers that were refractory to 5-FU/LV containing chemotherapy. However, disease stabilization was seen in a significant number of patients. PMID- 15342668 TI - Diagnosis of peripheral nerve sheath tumors around the pelvis. AB - OBJECTIVE: To distinguish between benign and malignant peripheral nerve sheath tumors around the pelvis. METHODS: A retrospective study of 30 patients with benign and malignant peripheral nerve sheath tumors located around the pelvis was performed. Clinical, imaging and histological features of 19 benign and 11 malignant peripheral nerve sheath tumors around the pelvis were reviewed retrospectively. RESULTS: Nearly all patients exhibited pain at presentation in cases involving both benign and malignant tumors. Although tumor size, duration of symptoms and presence of sensory disturbance possessed little value in differential diagnosis, severe motor weakness was observed exclusively in patients presenting with malignant tumors. On CT or MRI, central enhancement was apparent in 11 of the 19 benign tumors; in contrast, central enhancement was evident in one of the 11 malignant tumors. Fine needle aspiration cytology was performed in 11 tumors; correct diagnosis was achieved in four tumors. Core needle biopsy was performed in five tumors, all of which were correctly diagnosed with no neurological deficits. Immunohistochemically, all benign tumors were diffusely positive for S-100 protein, whereas malignant tumors were negative or focally positive for S-100 protein. Ki-67 index was less than 4% in all benign tumors; additionally, this index was 7-36% in malignant tumors. CONCLUSION: Central enhancement pattern on imaging studies strongly suggests a benign tumor; in contrast, severe motor weakness suggests malignant lesions. Core needle biopsy was reliable with respect to preoperative diagnosis. PMID- 15342669 TI - Prostate stem cell antigen (PSCA) expression in human prostate cancer tissues: implications for prostate carcinogenesis and progression of prostate cancer. AB - OBJECTIVE: Prostate stem cell antigen (PSCA) is a recently defined homolog of the Thy-1/Ly-6 family of glycosylphosphatidylinositol (GPI)-anchored cell surface antigens. The objective of the present study was to examine the expression status of PSCA protein and mRNA in clinical specimens of human prostate cancer (PCa) and to validate it as a potential molecular target for diagnosis and treatment of PCa. METHODS: Immunohistochemical (IHC) and in situ hybridization (ISH) analyses of PSCA expression were simultaneously performed on paraffin-embedded sections of 20 benign prostatic hyperplasia (BPH), 20 prostatic intraepithelial neoplasm (PIN) and 48 prostate cancer (PCa) tissues, including 9 androgen-independent prostate cancers. The level of PSCA expression was semiquantitatively scored by assessing both the percentage and intensity of PSCA-positive staining cells in the specimens. We then compared the PSCA expression between BPH, PIN and PCa tissues and analyzed the correlations of PSCA expression level with pathological grade, clinical stage and progression to androgen-independence in PCa. RESULTS: In BPH and low grade PIN, PSCA protein and mRNA staining were weak or negative and less intense and uniform than that observed in high grade PIN (HGPIN) and PCa. Moderate to strong PSCA protein and mRNA expression were noted in 8 of 11 (72.7%) HGPIN and in 40 of 48 (83.4%) PCa specimens examined by IHC and ISH analyses, and their statistical significance was compared with BPH (20%) and low grade PIN (22.2%) specimens (P < 0.05). The expression level of PSCA increased with a higher Gleason grade, advanced stage and progression to androgen independence (P < 0.05). In addition, IHC and ISH staining revealed a high degree of correlation between PSCA protein and mRNA overexpression. CONCLUSIONS: Our data demonstrate that PSCA as a new cell surface marker is overexpressed in a majority of cases of human PCa. PSCA expression correlates positively with adverse tumor characteristics, such as increasing pathological grade (poor cell differentiation), worsening clinical stage and androgen-independence and speculatively with prostate carcinogenesis. PSCA may possess prognostic utility and may be a promising molecular target for diagnosis and treatment of PCa. PMID- 15342670 TI - Efficacy of modest dose irradiation in combination with long-term endocrinal treatment for high-risk prostate cancer: a preliminary report. AB - BACKGROUND: Although radiotherapy in combination with endocrinal manipulation has been identified as an effective treatment for patients with high-risk prostate cancer, the optimal dose for locoregional control of prostate cancer in combination with hormonal therapy has not yet been determined. METHODS: The efficacy of modest doses of irradiation (60-62 Gy) combined with long-term endocrinal treatment for patients with high-risk prostate cancer (defined as a pretreatment prostate-specific antigen (PSA) level greater than 20 ng/ml or a Gleason's score of 8-10 or T3-T4 disease) was analyzed in 60 Japanese patients. The patients included in this study had received radical radiotherapy with long term endocrinal manipulation in the period between 1993 and 2000. The median age of the patients was 70 years (range, 56-83). Neoadjuvant hormonal therapy with a median duration of 3.9 months was performed prior to radiotherapy, and hormonal therapy was continued until recurrence. A median dose of 61.4 Gy (range, 44-71.4) was delivered to the prostate. Pelvic node irradiation was performed in 49 patients (81.6%). RESULTS: After a median follow-up period of 28.5 months, the overall survival, cause-specific survival and biochemical relapse-free survival at 3 years were 94.4%, 96% and 89.8%, respectively. Local failure was observed in one patient, distant metastases were observed in three patients and a late toxic effect greater than Grade 2 was not observed in any patients. CONCLUSIONS: This study, though preliminary due to a short-term follow-up period, reveals the possibility that modest doses of irradiation combined with long-term endocrinal treatment could be an effective means of achieving excellent local control of high-risk prostate cancer. PMID- 15342671 TI - Picture archiving and communication system introduced to a new Japanese cancer center hospital. AB - Picture archiving and communication systems (PACS) have been widely introduced as a credible alternative to the traditional film-based radiological service. Shizuoka Cancer Center Hospital (SCCH) introduced PACS and hospital information systems as an alternative to the film- and traditional paper-based hospital system at the opening of the hospital. Using PACS, whole images and their reports can be available at any nearby terminal. Users can select the display format for monitor viewing on the bases of individual preference and navigate through cases by buttons and rollerballs on the mouse with easy handling and quick response. Most clinicians in SCCH evaluated such medical circumstances well. Filmless image management systems will become popular in all hospitals in the near feature. All staff in each hospital should investigate the merits and demerits of this system and how to introduce it effectively. PMID- 15342672 TI - Scintimammographic findings of phyllodes tumor of the breast in a double-phase study with Tc-99m (V)DMSA and Tc-99m MIBI: a case report. AB - We present a case of phyllodes tumor of the breast in a 78-year-old woman evaluated with Tc-99m (V)DMSA and Tc-99m MIBI scintimammography, acquired in separate sessions (10 and 60 min post-injection). Tc-99m (V)DMSA was accumulated intensely within the mass [tumor to background ratio (T/B) >3], whereas Tc-99m MIBI had significantly lower uptake (T/B 1.9). Histology revealed a phyllodes tumor (maximum diameter 15 cm) and approximately three mitoses over 10 fields of view (FOV) x40, foci of epithelial hyperplasia and apocrine metaplasia. Stromal Ki-67 expression was 7%. The tumor was considered to be benign and the patient underwent mastectomy. One year later the patient presented with local malignant recurrence of the disease with over 15 mitoses per 10 FOV. Tc-99m (V)DMSA seems to have an advantage over Tc-99m MIBI in detecting mesenchymal tumors with unforeseen biological behavior and Ki-67 over-expression, such as phyllodes tumors, even with primary negative histological findings. PMID- 15342673 TI - Mortality trend of mouth and pharynx, esophagus, stomach, larynx and lung cancer in Japan by birth cohort. PMID- 15342674 TI - A review of CETP and its relation to atherosclerosis. AB - Although the atheroprotective role of HDL cholesterol (HDL-c) is well documented, effective therapeutics to selectively increase plasma HDL-c levels are not yet available. Recent progress in unraveling human HDL metabolism has fuelled the development of strategies to decrease the incidence and progression of coronary artery disease (CAD) by raising HDL-c. In this quest for novel drugs, cholesteryl ester transfer protein (CETP) represents a pivotal target. The role of this plasma protein in HDL metabolism is highlighted by the discovery that genetic CETP deficiency is the main cause of high HDL-c levels in Asian populations. The use of CETP inhibitors to effectively increase HDL-c concentration in humans was recently published and data with regard to the effect on human atherosclerosis are expected shortly. This review discusses the potential of CETP inhibitors to protect against atherosclerosis in the context of the current knowledge of CETP function in both rodents and humans. PMID- 15342675 TI - Phospholipids modify substrate binding and enzyme activity of human cytochrome P450 27A1. AB - Cytochrome P450 27A1 (P450 27A1) is an important metabolic enzyme involved in bile acid biosynthesis and the activation of vitamin D3 in mammals. Recombinant P450 27A1 heterologously expressed in Escherichia coli was found to be copurified with phospholipids (PLs). The PL content varied in different preparations and was dependent on the purification protocol. A link between the increased amounts of PLs and deterioration of the enzyme substrate binding properties was also observed. Tandem negative ionization mass spectrometry identified phosphatidylglycerol (PG) as the major PL copurified with P450 27A1. Subsequent reconstitution of P450 into exogenous PG vesicles assessed the effect of this contamination on substrate binding and enzyme activity. Two other PLs, phosphatidylethanolamine (PE) and phosphatidylserine (PS), were also tested. PG and PE increased the Kd for 5beta-cholestane-3alpha,7alpha,12alpha-triol and cholesterol binding, whereas PS had no effect on either substrate binding. PG and PE did not significantly alter 5beta-cholestane-3alpha,7alpha,12alpha-triol hydroxylase activity and even stimulated cholesterol hydroxylase activity. PS inhibited 5beta-cholestane-3alpha,7alpha,12alpha-triol hydrolyase activity and had no effect on cholesterol hydroxylase activity. Our study shows the potential for PLs to regulate the activity of P450 27A1 in vivo and alter the amount of cholesterol degraded through the "classical" and "alternative" bile acid biosynthetic pathways. PMID- 15342676 TI - Hydrophobic sequences target and anchor perilipin A to lipid droplets. AB - Perilipins regulate triacylglycerol storage and hydrolysis in adipocytes. The central 25% of the perilipin A sequence, including three hydrophobic sequences (H1, H2, and H3) and an acidic region, targets and anchors perilipins to lipid droplets. Thus, we hypothesized that H1, H2, and H3 are targeting and anchoring motifs. We now show that deletion of any single hydrophobic sequence or combinations of H1 and H3 or H2 and H3 does not prevent targeting of the mutated perilipin to lipid droplets. In contrast, mutated perilipin lacking H1 and H2 showed reduced targeting, whereas perilipin lacking H1, H2, and H3 targeted poorly to lipid droplets; thus, H3 is a weak targeting signal and either H1 or H2 is required for optimal targeting. Complete elimination of perilipin targeting was observed only when all three hydrophobic sequences were deleted in combination with either the acidic region or N-terminal sequences predicted to form amphipathic beta-strands. Unlike intact perilipin A, mutated perilipin lacking either H1 and H2 or H1, H2, and H3 was released from lipid droplets after alkaline carbonate treatment, suggesting that these forms are loosely associated with lipid droplets. The three hydrophobic sequences play a major role in targeting and anchoring perilipins to lipid droplets. PMID- 15342677 TI - Validation of deuterium-labeled fatty acids for the measurement of dietary fat oxidation during physical activity. AB - Measurement of 13C-labeled fatty acid oxidation is hindered by the need for acetate correction, measurement of the rate of CO2 production in a controlled environment, and frequent collection of breath samples. The use of deuterium labeled fatty acids may overcome these limitations. Herein, d31-palmitate was validated against [1-13C]palmitate during exercise. Thirteen subjects with body mass index of 22.9 +/- 3 kg/m2 and body fat of 19.6 +/- 11% were subjected to 2 or 4 h of exercise at 25% maximum volume oxygen consumption (VO2max). The d31 palmitate and [1-13C] palmitate were given orally in a liquid meal at breakfast. The d3-acetate and [1-13C]acetate were given during another visit for acetate sequestration correction. Recovery of d31-palmitate in urine at 9 h after dose was compared with [1-13C] palmitate recovery in breath. Cumulative recovery of d31-palmitate was 10.6 +/- 3% and that of [1-13C]palmitate was 5.6 +/- 2%. The d3 acetate and [1-13C]acetate recoveries were 85 +/- 4% and 54 +/- 4%, respectively. When [1-13C]acetate recovery was used to correct 13C data, the average recovery differences were 0.4 +/- 3%. Uncorrected d31-palmitate and acetate-corrected [1 13C]palmitate were well correlated (y=0.96x + 0; P <0.0001) when used to measure fatty acid oxidation during exercise. Thus, d31-palmitate can be used in outpatient settings as it eliminates the need for acetate correction and frequent sampling. PMID- 15342678 TI - Apolipoprotein A-I, A-II, and VLDL-B-100 metabolism in men: comparison of a low fat diet and a high-monounsaturated fatty acid diet. AB - The impact of a low-fat diet and a high-MUFA diet on apolipoprotein A-I (apoA-I), apoA-II, and VLDL-apoB-100 metabolism in conditions of unrestricted (ad libitum) energy intake was compared in 65 men randomly assigned to one of two predefined experimental diets. A subsample of 18 men participated in the kinetic study. Before and after the 6-7 week dietary intervention, kinetic subjects received a primed-constant infusion of [5,5,5-2H3]L-leucine for 12 h under feeding conditions. ApoA-I production rate (PR; -31.5%; P <0.001) and fractional catabolic rate (FCR; -24.3%; P <0.05) were significantly decreased after the low fat diet. These changes in apoA-I PR and FCR with the low-fat diet were also significantly different from those observed with the high-MUFA diet (P <0.01 and P <0.05, respectively). ApoA-II FCR was significantly increased in the high-MUFA group only. No significant within- or between-diet difference was found in VLDL apoB-100 PR or FCR. These results emphasize the differential impact of the low fat diet and high-MUFA diet on HDL metabolism. PMID- 15342679 TI - Active and low-active forms of serum phospholipid transfer protein in a normal Finnish population sample. AB - Human serum phospholipid transfer protein (PLTP) exists as a catalytically active (HA-PLTP) and a low-active (LA-PLTP) form. In this study, the association of PLTP activity and the concentrations of both forms with lipid and carbohydrate parameters were investigated. In a random Finnish population sample, serum PLTP concentration (n=250) was 6.56 +/- 1.45 mg/l, the mean lipoprotein-independent (PLTPexo) phospholipid transfer activity was 6.59 +/- 1.66 micromol/ml/h, and the mean lipoprotein-dependent (PLTPendo) activity was 1.37 +/- 0.29 micromol/ml/h. Of the serum PLTP concentration, approximately 46% was in a catalytically active form. HA-PLTP concentration correlated positively with serum PLTPexo activity (r=0.380, P <0.001), HDL cholesterol (r=0.291, P <0.001), and apolipoprotein A-I (r=0.187, P <0.01). Of the potential regulatory factors for PLTP, apolipoprotein E showed a weak positive correlation with serum PLTPexo (r=0.154, P <0.05) and PLTPendo (r=0.192, P <0.01) activity but not with PLTP concentration. Weak associations were also observed between PLTP parameters and determinants of glucose homeostasis (glucose, insulin, and homeostasis model assessment for insulin resistance). The present data on PLTP activity and concentration reveal novel connections of the two PLTP forms to lipid and carbohydrate metabolism. PMID- 15342680 TI - Novel lipoidal derivatives of pregnenolone and dehydroepiandrosterone and absence of their sulfated counterparts in rodent brain. AB - A new sample preparation method coupled to GC-MS analysis was developed and validated for quantification of sulfate esters of pregnenolone (PREG-S) and dehydroepiandrosterone (DHEA-S) in rat brain. Using a solid-phase extraction recycling protocol, the results show that little or no PREG-S and DHEA-S (<1 pmol/g) is present in rat and mouse brain. These data are in agreement with studies in which steroid sulfates were analyzed without deconjugation. We suggest that the discrepancies between analyses with and without deconjugation are caused by internal contamination of brain extract fractions, supposed to contain steroid sulfates, by lipoidal forms of PREG and DHEA (L-PREG and L-DHEA, respectively). These derivatives can be acylated very efficiently with heptafluorobutyric anhydride and triethylamine, and their levels in rodent brain (approximately 1 nmol/g) are much higher than those of their unconjugated counterparts. They are distinct from fatty acid esters, and preliminary data do not favor structures such as sulfolipids or sterol peroxides. Noncovalent interactions between steroids and proteolipidic elements, such as lipoproteins, could account for some experimental data. Given their abundance in rodent brain, the structural characterization and biological functions of L-PREG and L-DHEA in the central nervous system merit considerable attention. PMID- 15342681 TI - Fatty acid binding proteins stabilize leukotriene A4: competition with arachidonic acid but not other lipoxygenase products. AB - Leukotriene A(4) (LTA(4)) is a chemically reactive conjugated triene epoxide product derived from 5-lipoxygenase oxygenation of arachidonic acid. At physiological pH, this reactive compound has a half-life of less than 3 s at 37 degrees C and approximately 40 s at 4 degrees C. Regardless of this aqueous instability, LTA(4) is an intermediate in the formation of biologically active leukotrienes, which can be formed through either intracellular or transcellular biosynthesis. Previously, epithelial fatty acid binding protein (E-FABP) present in RBL-1 cells was shown to increase the half-life of LTA(4) to approximately 20 min at 4 degrees C. Five FABPs (adipocyte FABP, intestinal FABP, E-FABP, heart/muscle FABP, and liver FABP) have now been examined and also found to increase the half-life of LTA(4) at 4 degrees C to approximately 20 min with protein present. Stabilization of LTA(4) was examined when arachidonic acid was present to compete with LTA(4) for the binding site on E-FABP. Arachidonate has an apparent higher affinity for E-FABP than LTA(4) and was able to completely block stabilization of the latter. When E-FABP is not saturated with arachidonate, FABP can still stabilize LTA(4). Several lipoxygenase products, including 5-hydroxyeicosatetraenoic acid, 5,6-dihydroxyeicosatetraenoic acid, and leukotriene B(4), were found to have no effect on the stability of LTA(4) induced by E-FABP even when present at concentrations 3-fold higher than LTA(4). PMID- 15342682 TI - Demonstration of reverse fatty acid transport from rat cardiomyocytes. AB - Fatty acids flow from adipocytes to nonadipose tissues during fasting and exercise and normally are fully oxidized. To determine if nonadipose tissues can export unoxidized FA when FA influx exceeds oxidation, neonatal cardiomyocytes were cultured in 1 microCi (14)C-palmitate in the presence of etomoxir to block oxidation. The cells took up and stored 25% of the radioactivity as (14)C triacylglycerol in 12 h, but 4.5% of the label was released in 3 h and comigrated with (14)C-palmitate. Both uptake and release of radioactivity were increased by insulin and reduced by the nonspecific inhibitors of FA transporters phloretin and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). Perfused hearts from etomoxir-treated lean rats released 221 +/- 59 nmol/10 min of FA. Hearts from high-fat-fed lean rats released 366 +/- 172 nmol/10 min (P < 0.05). Hearts from obese rats released 744 +/- 260 and 1,578 +/- 630 nmol/10 min at 8 and 12 weeks of age, respectively. Perfusion with insulin increased FA release by 32%. In vitro and ex vivo findings suggest that nonadipose tissues such as myocardium can export FA when the unoxidized lipid content is excessive. PMID- 15342683 TI - Hypercholesterolemia in ENU-induced mouse mutants. AB - Hypercholesterolemia is caused by multiple environmental factors and genetic predispositions, and plays an important role in the development and pathogenesis of various human diseases. In this study, we aimed to establish randomly mutant mouse lines showing hypercholesterolemia for their further use in the detection of novel causative alleles. In the Munich ENU Mouse Mutagenesis Project, clinical chemistry blood analysis was performed on more than 15,000 G1 mice and 230 G3 pedigrees of chemically mutagenized mice to detect dominant and recessive mutations leading to an increased plasma total cholesterol level. Using inbred C3HeB/FeJ mice we identified more than 100 animals consistently showing hypercholesterolemia. Transmission of the altered phenotype to the subsequent generations led to the production of nine hypercholesterolemic lines. A single line showed further obvious deviations in the analysis of additional clinical chemistry blood parameters. Thus, the lines produced will contribute to the search for alleles that selectively cause primary hypercholesterolemia. PMID- 15342684 TI - Effect of increasing the expression of cholesterol transporters (StAR, MLN64, and SCP-2) on bile acid synthesis. AB - There are two major pathways of bile acid synthesis: the "neutral" pathway, initiated by highly regulated microsomal cholesterol 7alpha-hydroxylase (CYP7A1), and an "alternative" pathway, initiated by mitochondrial sterol 27-hydroxylase (CYP27A1). In hepatocyte cultures, overexpression of CYP7A1 increases bile acid synthesis by >8-fold. However, overexpression of CYP27A1 in hepatocytes only increases it by 1.5-fold, suggesting that additional rate-limiting steps must be involved in the regulation of this pathway. The effects of intracellular cholesterol transport proteins on bile acid synthesis have been investigated in the current study. Under culture conditions in which the neutral pathway was inactive, selective overexpression of the gene encoding steroidogenic acute regulatory protein (StAR), MLN64 (StAR homolog protein), and sterol carrier protein-2 (SCP-2) led to 5.7-, 1.2-, and 1.7-fold increases, respectively, in the rates of bile acid synthesis in primary rat hepatocytes. Surprisingly, co overexpression of MLN64 with StAR, SCP-2, or CYP7A1 blunted the upregulated bile acid synthesis by 48, 47, and 45%, respectively. These results suggest that MLN64, in its full-length form, is not responsible for the transport of cholesterol to the mitochondria or the endoplasmic reticulum, where CYP27A1 or CYP7A1 is located, respectively. PMID- 15342685 TI - Farnesoid X receptor represses hepatic lipase gene expression. AB - The farnesoid X receptor (FXR) is a nuclear receptor that regulates gene expression in response to bile acids (BAs). FXR plays a central role in BA, cholesterol, and lipoprotein metabolism. Here, we identify HL, an enzyme involved in the metabolism of remnant and high density lipoproteins, as a novel FXR regulated gene. The natural FXR ligand, chenodeoxycholic acid (CDCA), downregulates HL gene expression in a dose- and time-dependent manner in human hepatoma HepG2 cells. The nonsteroidal synthetic FXR agonist GW4064 also decreases HL mRNA levels in HepG2 cells and in primary human hepatocytes. Moreover, the decrease of HL mRNA levels after treatment with FXR agonists was associated with a significant decrease in secreted enzymatic activity. In addition, FXR-specific gene silencing using small interfering RNAs demonstrated that CDCA- and GW4064-mediated downregulation of HL transcript levels occurs via an FXR-dependent mechanism. Finally, using transient transfection experiments, it is shown that FXR represses transcriptional activity of a reporter driven by the 698/+13 bp human HL promoter. Taken together, these results identify HL as a new FXR-regulated gene in human liver cells. In view of the role of HL in plasma lipoprotein metabolism, our results further emphasize the central role of FXR in lipid homeostasis. PMID- 15342686 TI - Purified human serum PON1 does not protect LDL against oxidation in the in vitro assays initiated with copper or AAPH. AB - Purified serum paraoxonase (PON1) had been shown to attenuate the oxidation of LDL in vitro. We critically reevaluated the antioxidant properties of serum PON1 in the in vitro assays initiated with copper or the free radical generator 2,2' azobis-2-amidinopropane hydrochloride (AAPH). The antioxidant activity of different purified PON1 preparations did not correlate with their arylesterase (AE), lactonase, or phospholipase A2 activities or with the amounts of detergent or protein. Dialysis of three of these preparations resulted in a 30-40% loss of their AE activities but in a complete loss of their antioxidant activities. We also followed the distribution of the antioxidant activity during human serum PON1 purification by two purification methods. The antioxidant activity of the anion-exchange chromatography fractions did not copurify with PON1 using either method and could largely be accounted for by the "antioxidant" activity of the detergent present. In conclusion, using the copper or AAPH in vitro assays, no PON1-mediated antioxidant activity was detected, suggesting that the removal of PON1 from its natural environment may impair its antioxidative activity and that this assay with highly purified PON1 may be an inappropriate method with which to study the antioxidative properties of the enzyme. PMID- 15342687 TI - Stanol esters decrease plasma cholesterol independently of intestinal ABC sterol transporters and Niemann-Pick C1-like 1 protein gene expression. AB - Possible mechanisms for the cholesterol-lowering effects of plant stanol esters were addressed by feeding hamsters diets containing stanol esters, cholesterol, or cholestyramine/lovastatin. ABCA1, ATP binding cassette G1 (ABCG1), ABCG5, ABCG8, and Niemann-Pick C1-like 1 (NPC1L1) mRNA levels were then estimated in duodenum, jejunum, and ileum. Plasma cholesterol was decreased by 36% and 94% in animals fed stanol esters and cholestyramine/lovastatin, respectively. Cholesterol feeding increased plasma cholesterol by 2.5-fold. Plasma plant sterols were unchanged by stanol ester feeding but became undetectable by feeding cholestyramine/lovastatin. Cholesterol and stanols accumulated in enterocytes of animals fed cholesterol and stanol esters, respectively. ABCG5 and ABCG8 mRNA levels were decreased by stanol esters and cholestyramine/lovastatin. Cholesterol feeding markedly increased ABCA1 and ABCG1 expression and modestly increased ABCG5/ABCG8. NPC1L1 mRNA was not significantly altered by any of the diets. ABCG1, ABCG5, ABCG8, and NPC1L1 mRNAs were highest in cells of the upper villus, whereas ABCA1 mRNA was highest in cells of the lower villus. The results suggest that cholesterol lowering effect of stanol esters is unrelated to changes in mRNA levels of intestinal ABC sterol transporters or NPC1L1. Cholesterol flux regulates ABC expression but not NPC1L1. The different localization of ABCA1 suggests a different function for this protein than for ABCG1, ABCG5, ABCG8, and NPC1L1. PMID- 15342688 TI - Influence of the APOA5 locus on plasma triglyceride, lipoprotein subclasses, and CVD risk in the Framingham Heart Study. AB - Several polymorphisms in the APOA5 gene have been associated with increased plasma triglyceride (TG) concentrations. However, associations between APOA5 and lipoprotein subclasses, remnant-like particles (RLPs), and cardiovascular disease (CVD) risk have been less explored. We investigated associations of five APOA5 single-nucleotide polymorphisms (SNPs; -1131T>C, -3A>G, 56C>G IVS3+ 476G>A, and 1259T>C) with lipoprotein subfractions and CVD risk in 1,129 men and 1,262 women participating in the Framingham Heart Study. Except for the 56C>G SNP, the other SNPs were in significant linkage disequilibria, resulting in three haplotypes (11111, 22122, and 11211) representing 98% of the population. SNP analyses revealed that the -1131T>C and 56C>G SNPs were significantly associated with higher plasma TG concentrations in both men and women. For RLP and lipoprotein subclasses, we observed gender-specific association for the -1131T>C and 56C>G SNPs. Female carriers of the -1131C allele had higher RLP concentrations, whereas in males, significant associations for RLPs were observed for the 56G allele. Moreover, haplotype analyses confirmed these findings and revealed that the 22122 and 11211 haplotypes exhibited different associations with HDL cholesterol concentrations. In women, the -1131C allele was associated with a higher hazard ratio for CVD (1.85; 95% confidence interval, 1.03-3.34; P = 0.04), in agreement with the association of this SNP with higher RLPs. PMID- 15342689 TI - Overexpression of apoC-III produces lesser hypertriglyceridemia in apoB-48-only gene-targeted mice than in apoB-100-only mice. AB - The adaptive value of apolipoprotein B-48 (apoB-48), the truncated form of apoB produced by the intestine, in lipid metabolism remains unclear. We crossed human apoC-III transgenic mice with mice expressing either apoB-48 only (apoB48/48) or apoB-100 only (apoB100/100). Cholesterol levels were higher in apoB48/48 mice than in apoB100/100 mice but triglyceride levels were similar. Lipid levels were increased by the apoC-III transgene. However, triglyceride levels were significantly higher in apoB100/100C-III than in apoB48/48C-III mice (895 +/- 395 mg/dl vs. 690 +/- 252 mg/dl; P <0.01), whereas cholesterol levels were higher in the apoB48/48C-III mice than in apoB100/100C-III (144 +/- 35 mg/dl vs. 94 +/- 30 mg/dl; P <0.00001). Triglyceride clearance from VLDL was impaired to a greater extent in apoB100/100C-III vs. apoB100/100 mice than in apoB48/48C-III vs. apoB48/48 mice. Triglyceride secretion rates were no different in apoC-III transgenic mice than in their nontransgenic littermates. ApoB-48 triglyceride rich lipoproteins were more resistant to the triglyceride-increasing effects of apoC-III but appeared more sensitive to the remnant clearance inhibition. Our findings support a coordinated role for apoB-48 in facilitating the delivery of dietary triglycerides to the periphery. Consistent with such a mechanism, glucose levels were significantly higher in apoB48/48 mice vs. apoB100/100 mice, perhaps on the basis of metabolic competition. PMID- 15342690 TI - Role of N-linked glycosylation in the secretion and activity of endothelial lipase. AB - Human endothelial lipase (EL), a member of the triglyceride lipase gene family, has five potential N-linked glycosylation sites, two of which are conserved in both lipoprotein lipase and hepatic lipase. Reduction in molecular mass of EL after treatment with glycosidases and after treatment of EL-expressing cells with the glycosylation inhibitor tunicamycin demonstrated that EL is a glycosylated protein. Each putative glycosylation site was examined by site-directed mutagenesis of the asparagine (Asn). Mutation of Asn-60 markedly reduced secretion and slightly increased specific activity. Mutation of Asn-116 did not influence secretion but increased specific activity. In both cases, this resulted from decreased apparent K(m) and increased apparent V(max). Mutation of Asn-373 did not influence secretion but significantly reduced specific activity, as a result of a decrease in apparent V(max). Mutation of Asn-471 resulted in no reduction in secretion or specific activity. Mutation of Asn-449 resulted in no change in secretion, activity, or molecular mass, indicating that the site is not utilized. The ability of mutants secreted at normal levels to mediate bridging between LDL and cell surfaces was examined. The Asn-373 mutant demonstrated a 3 fold decrease in bridging compared with wild-type EL, whereas Asn-116 and Asn-471 were similar to wild-type EL. PMID- 15342691 TI - Requirement of PPARalpha in maintaining phospholipid and triacylglycerol homeostasis during energy deprivation. AB - The peroxisome proliferator-activated receptor alpha (PPARalpha) has been implicated as a key control of fatty acid catabolism during the cellular fasting. However, little is known regarding changes of individual fatty acids in hepatic triacylglycerol (TG) and phospholipid (PL) as a result of starvation. In the present work, the effects of 72 h fasting on hepatic TG and PL fatty acid profiles in PPARalpha-null (KO) mice and their wild-type (WT) counterparts were investigated. Our results indicated that mice deficient in PPARalpha displayed hepatomegaly and hypoketonemia following 72 h starvation. Histochemical analyses revealed that severe fatty infiltration was observed in the livers of KO mice under fasted conditions. Furthermore, 72 h fasting resulted in a 2.8-fold higher accumulation of hepatic TG in KO mice than in WT mice fasted for the same length of time. Surprisingly, the total hepatic PL contents in fasted KO mice decreased by 45%, but no significant change in hepatic PL content was observed in WT mice following starvation. Gas chromatographic analysis indicated that KO mice were deprived of arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acids during fasting. Taken together, these results show that PPARalpha plays an important role in regulation of fatty acid metabolism as well as phospholipid homeostasis during energy deprivation. PMID- 15342692 TI - The genetics of strabismus. AB - Strabismus (misalignment of the eyes; also known as "squint") comprises a common heterogeneous group of disorders characterised by a constant or intermittent ocular deviation often associated with amblyopia (uniocular failure of normal visual development) and reduced or absent binocular vision. The associated poor cosmetic appearance may also interfere with social and psychological development. Extensive twin and family studies suggest a significant genetic component to the aetiology of strabismus. The complexity of the molecular basis of strabismus is now beginning to be elucidated with the identification of genetic loci and disease causing genes. Currently greater insights have been gained into the incomitant subtype (differing magnitude of ocular misalignment according to direction of gaze), whereas less is known about the pathogenesis of the more common childhood concomitant strabismus. It is hoped that a greater understanding of the molecular genetics of these disorders will lead to improved knowledge of disease mechanisms and ultimately to more effective treatment. The aim of this paper is to review current knowledge of the molecular genetics of both incomitant and concomitant strabismus. PMID- 15342693 TI - CYP1B1 mutations in French patients with early-onset primary open-angle glaucoma. AB - INTRODUCTION: Primary open-angle glaucoma (POAG) is a leading cause of visual impairment worldwide and a complex genetic disorder that affects mostly adults. Mutations in the MYOCILIN (MYOC) and OPTINEURIN genes account for rare forms with a Mendelian inheritance and for <5% of all POAG cases. The CYP1B1 gene, a member of the cytochrome P450 gene family, is a major cause of primary congenital glaucoma (PCG), a rare and severely blinding disease with recessive inheritance. However, CYP1B1 mutations have also been associated with cases of juvenile-onset glaucoma in some PCG families or shown to modify the age of onset of glaucoma linked to a MYOC mutation in a large family. OBJECTIVE: To investigate the role of CYP1B1 mutations in POAG predisposition, irrespective of the presence of a MYOC mutation. METHODS AND SUBJECTS: CYP1B1 coding region variation was characterised by denaturing high performance liquid chromatography (DHPLC) and sequencing in 236 unrelated French Caucasian POAG patients and 47 population matched controls. RESULTS: Eleven (4.6%) patients carried one or two mutated CYP1B1 gene(s) and no MYOC mutation. They showed juvenile or middle-age onset of disease (median age at diagnosis, 40 years, range 13-52), significantly earlier than in non-carrier patients. Apart from one, all mutations detected in POAG patients were previously associated with PCG. CONCLUSION: CYP1B1 mutations might pose a significant risk for early-onset POAG and might also modify glaucoma phenotype in patients who do not carry a MYOC mutation. PMID- 15342694 TI - Genomewide scan identifies susceptibility locus for dyslexia on Xq27 in an extended Dutch family. AB - CONTEXT: Dyslexia is a common disorder with a strong genetic component, but despite significant research effort, the aetiology is still largely unknown. OBJECTIVE: To identify loci contributing to dyslexia risk. METHODS: This was a genomewide linkage analysis in a single large family. Dutch families with at least two first degree relatives suffering from dyslexia participated in the study. Participants were recruited through an advertisement campaign in papers and magazines. The main outcome measure was linkage between genetic markers and dyslexia phenotype. RESULTS: Using parametric linkage analysis, we found strong evidence for a locus influencing dyslexia on Xq27.3 (multipoint lod = 3.68). Recombinations in two family members flanked an 8 cM region, comprising 11 currently confirmed genes. All four males carrying the risk haplotype had very low scores on the reading tests. The presentation in females was more variable, but 8/9 females carrying the risk haplotype were diagnosed dyslexic by our composite score, so we considered the putative risk allele to be dominant with reduced penetrance. Linkage was not found in an additional collection of affected sibling pairs. CONCLUSIONS: A locus influencing dyslexia risk is probably located between markers DXS1227 and DXS8091 on the X chromosome, closely situated to a locus indicated by a published genome scan of English sibling pairs. Although the locus may not be a common cause for dyslexia, the relatively small and gene poor region offers hope to identify the responsible gene. PMID- 15342695 TI - Association of a STAT 6 haplotype with elevated serum IgE levels in a population based cohort of white adults. AB - BACKGROUND: Several studies have shown linkage of chromosome 12q 13-24 with atopy related phenotypes. Among candidate genes in this region is STAT6 (signal transducer and activator of transcription), which is essential for Th2 cell differentiation, recruitment, and effector function. METHODS: We evaluated six polymorphisms of STAT6 for evidence of associations with serum IgE levels and atopic diseases in a population based cross sectional cohort of 1407 German adults. Genotyping was performed using the matrix assisted laser desorption ionisation-time of flight mass spectrometry method. Haplotypes were estimated using the SAS/Genetics module, and population-derived IgE percentiles (50% IgE>53 kU/l, 66% IgE>99 kU/l and 90% IgE>307 kU/l) were modelled as outcome variables in haplotype trend regression analysis. RESULTS: All polymorphisms were genotyped successfully. Haplotype reconstruction revealed 8/64 possible haplotypes, reaching estimated frequencies of 1% or more. One polymorphism in intron 2 (rs324011) showed a significant association with total serum IgE (p = 0.015). A STAT6 risk haplotype for elevated IgE showing odds ratios of 1.7 (p = 0.015) for IgE cut-off 100 kU/l, and 1.54 (p = 0.032), 1.6 (p = 0.025), and 2.54 (p = 0.007) for IgE percentiles 50%, 66%, and 90%, respectively was detected. The increased risk of this haplotype was confirmed by linear haplotype trend regression on log transformed IgE values (p = 0.007). Analysis further revealed a risk haplotype for specific sensitisation and a risk haplotype for asthma. CONCLUSION: The data indicate that genetic variants within STAT6 contribute significantly to IgE regulation and manifestation of atopic diseases. PMID- 15342696 TI - BRAF screening as a low-cost effective strategy for simplifying HNPCC genetic testing. AB - BACKGROUND: According to the international criteria for hereditary non-polyposis colorectal cancer (HNPCC) diagnostics, cancer patients with a family history or early onset of colorectal tumours showing high microsatellite instability (MSI-H) should receive genetic counselling and be offered testing for germline mutations in DNA repair genes, mainly MLH1 and MSH2. Recently, an oncogenic V600E hotspot mutation within BRAF, a kinase encoding gene from the RAS/RAF/MAPK pathway, has been found to be associated with sporadic MSI-H colon cancer, but its association with HNPCC remains to be further clarified. METHODS: BRAF-V600E mutations were analysed by automatic sequencing in colorectal cancers from 206 sporadic cases with MSI-H and 111 HNPCC cases with known germline mutations in MLH1 and MSH2. In addition, 45 HNPCC cases showing abnormal immunostaining for MSH2 were also analysed. RESULTS: The BRAF-V600E hotspot mutation was found in 40% (82/206) of the sporadic MSI-H tumours analysed but in none of the 111 tested HNPCC tumours or in the 45 cases showing abnormal MSH2 immunostaining. CONCLUSIONS: Detection of the V600E mutation in a colorectal MSI-H tumour argues against the presence of a germline mutation in either the MLH1 or MSH2 gene. Therefore, screening of these mismatch repair (MMR) genes can be avoided in cases positive for V600E if no other significant evidence, such as fulfilment of the strict Amsterdam criteria, suggests MMR associated HNPCC. In this context, mutation analysis of the BRAF hotspot is a reliable, fast, and low cost strategy which simplifies genetic testing for HNPCC. PMID- 15342697 TI - Array comparative genomic hybridisation analysis of boys with X linked hypopituitarism identifies a 3.9 Mb duplicated critical region at Xq27 containing SOX3. AB - INTRODUCTION: Array comparative genomic hybridisation (array CGH) is a powerful method that detects alteration of gene copy number with greater resolution and efficiency than traditional methods. However, its ability to detect disease causing duplications in constitutional genomic DNA has not been shown. We developed an array CGH assay for X linked hypopituitarism, which is associated with duplication of Xq26-q27. METHODS: We generated custom BAC/PAC arrays that spanned the 7.3 Mb critical region at Xq26.1-q27.3, and used them to search for duplications in three previously uncharacterised families with X linked hypopituitarism. RESULTS: Validation experiments clearly identified Xq26-q27 duplications that we had previously mapped by fluorescence in situ hybridisation. Array CGH analysis of novel XH families identified three different Xq26-q27 duplications, which together refine the critical region to a 3.9 Mb interval at Xq27.2-q27.3. Expression analysis of six orthologous mouse genes from this region revealed that the transcription factor Sox3 is expressed at 11.5 and 12.5 days after conception in the infundibulum of the developing pituitary and the presumptive hypothalamus. DISCUSSION: Array CGH is a robust and sensitive method for identifying X chromosome duplications. The existence of different, overlapping Xq duplications in five kindreds indicates that X linked hypopituitarism is caused by increased gene dosage. Interestingly, all X linked hypopituitarism duplications contain SOX3. As mutation of this gene in human beings and mice results in hypopituitarism, we hypothesise that increased dosage of Sox3 causes perturbation of pituitary and hypothalamic development and may be the causative mechanism for X linked hypopituitarism. PMID- 15342698 TI - A splice site mutation in the methyltransferase gene FTSJ1 in Xp11.23 is associated with non-syndromic mental retardation in a large Belgian family (MRX9). AB - Mental retardation is the most frequent cause of serious handicap in children and young adults. The underlying causes of this heterogeneous condition are both acquired and genetically based. A recently performed refinement of the linkage interval in a large Belgian family with mild to severe non-syndromic X linked mental retardation, classified as MRX9, revealed a candidate region of 11.3 Mb between markers DXS228 and DXS1204 on the short arm of the X chromosome. In order to identify the underlying disease gene in the MRX9 family, we established a gene catalogue for the candidate region and performed comprehensive mutation analysis by direct sequencing. A human homologue of the bacterial 23S rRNA methyltransferase Fstj, the FTSJ1 gene, is located within this region and displayed a sequence alteration in the conserved acceptor splice site of intron 3 (IVS3-2A>G) in all tested patients and carrier females of this family. In contrast, it was absent in all unaffected male family members tested. The mutation results in skipping of exon 4 and introduces a premature stop codon in exon 5, probably leading to a severely truncated protein. Our finding indicates that a protein, possibly associated with ribosomal stability, can be linked to X linked mental retardation (XLMR). PMID- 15342699 TI - Somatic NKX2-5 mutations as a novel mechanism of disease in complex congenital heart disease. AB - NKX2-5 is a pivotal transcription factor in heart development. Previous studies on lymphocytic DNA provided evidence of familial NKX2-5 gene mutations in cardiac malformations. Common mutations are rare in unrelated families. We analysed, by direct sequencing, the gene encoding NKX2-5 in the diseased heart tissues of 68 patients with complex congenital heart disease, focussing particularly on atrial, ventricular, and atrioventricular septal defects. We identified 35 non-synonymous NKX2-5 mutations in the diseased heart tissues of patients. These mutations were mainly absent in normal, for example, unaffected, heart tissue of the same patient, indicating the somatic nature and mosaicism of mutations. We also observed multiple mutations and multiple haplotypes, as well as mutations in Down's syndrome patients with cardiac malformations. Taken collectively, the above results suggest the somatic nature of NKX2-5 mutations associated with complex cardiac malformations. Somatic mutations in transcription factor genes of cardiac progenitor cells provide a novel mechanism of disease. PMID- 15342700 TI - Mild Wolf-Hirschhorn syndrome: micro-array CGH analysis of atypical 4p16.3 deletions enables refinement of the genotype-phenotype map. PMID- 15342701 TI - Identification of novel locus for autosomal dominant butterfly shaped macular dystrophy on 5q21.2-q33.2. PMID- 15342702 TI - An Alu-mediated partial SDHC deletion causes familial and sporadic paraganglioma. PMID- 15342703 TI - Novel locus on chromosome 12q22-q23.3 responsible for familial temporal lobe epilepsy associated with febrile seizures. PMID- 15342704 TI - Lamin A expression levels are unperturbed at the normal and mutant alleles but display partial splice site selection in Hutchinson-Gilford progeria syndrome. PMID- 15342707 TI - OPA3 gene mutations responsible for autosomal dominant optic atrophy and cataract. PMID- 15342708 TI - Single nucleotide polymorphism (SNP) analysis of mouse quantitative trait loci for identification of candidate genes. PMID- 15342709 TI - Transmission disequilibrium test of stromelysin-1 gene variation in relation to Crohn's disease. PMID- 15342710 TI - SALL4 deletions are a common cause of Okihiro and acro-renal-ocular syndromes and confirm haploinsufficiency as the pathogenic mechanism. PMID- 15342711 TI - Mutation screening of the BARD1 gene: evidence for involvement of the Cys557Ser allele in hereditary susceptibility to breast cancer. PMID- 15342712 TI - Stimulus dependence of disparity coding in primate visual area V4. AB - Disparity tuning in visual cortex has been shown using a variety of stimulus types that contain stereoscopic depth cues. It is not known whether different stimuli yield similar disparity tuning curves. We studied whether cells in visual area V4 of the macaque show similar disparity tuning profiles when the same set of disparity values were tested using bars or dynamic random dot stereograms, which are among the most commonly used stimuli for this purpose. In a majority of V4 cells (61%), the shape of the disparity tuning profile differed significantly for the two stimulus types. The two sets of stimuli yielded statistically indistinguishable disparity tuning profiles for only a small minority (6%) of V4 cells. These results indicate that disparity tuning in V4 is stimulus-dependent. Given the fact that bar stimuli contain two-dimensional (2-D) shape cues, and the random dot stereograms do not, our results also indicate that V4 cells represent 2-D shape and binocular disparity in an interdependent fashion, revealing an unexpected complexity in the analysis of depth and three-dimensional shape. PMID- 15342713 TI - A comparison of neuron response properties in areas A1 and CM of the marmoset monkey auditory cortex: tones and broadband noise. AB - The purpose of this study was to compare response properties of two adjacent areas of the marmoset monkey auditory cortex. Multiunit responses to 50 ms tones and broadband noise bursts (BBN) were recorded in the core area, A1, and the caudomedial belt area, CM, of ketamine-anesthetized animals. Neurons in A1 and CM exhibited robust low-threshold short-latency responses to BBN and tones, whereas neurons in adjoining lateral belt areas were poorly responsive or unresponsive to tones and noise. Except for a population of broadly tuned units in CM, the characteristic frequency (CF) could be determined for all recording sites in A1 and CM. Both areas were tonotopically organized and shared a high CF border. Whereas the tonotopic gradient in A1 was smooth and continuous across the field, the gradient in CM was discontinuous, and the intermediate CF range was underrepresented. For BBN stimuli, rate level functions were largely monotonic in A1 and CM. Response profiles were also similar in both areas. As a population, neurons in CM were distinguished from A1 by significantly shorter response latencies, lower thresholds, and broader tuning bandwidth at higher intensities. The results indicated that, while A1 and CM represent anatomically and physiologically distinct areas, their response profiles under anesthesia overlapped considerably compared with the lateral belt areas. Therefore refinements of current models of the primate auditory cortex may be needed to account for differences in organization among the auditory belt areas. PMID- 15342714 TI - The speed of auditory low-side suppression. AB - The nonlinear cochlear phenomenon of two-tone suppression is known to be very fast, but precisely how fast is unknown. We studied the timing of low-side suppression in the auditory nerve of the cat using multitone complexes as auditory stimuli. An evalution of the group delays of the responses to these complexes allowed us to measure the timing of the responses with sub-millisecond accuracy for a large number of fibers with characteristic frequencies (CFs) between 2 and 40 kHz. In particular, we measured the delays with which the same below-CF tone complexes affected the response either as an excitor (when presented alone) or as a suppressor (when combined with a CF probe). For CFs <10 kHz, we found that the delay of suppression was larger than the delay of excitation by several hundred microseconds. The difference between the delay of suppression and that of excitation decreased with increasing CF, becoming negligible for CFs >15 kHz. The results are analyzed in terms of traveling-wave delays and a purported cochlear gain control. The data suggest that suppression originates from a gain-control mechanism with an integration time in the order of two cycles of CF. PMID- 15342715 TI - Effects of local nicotinic activation of the superior colliculus on saccades in monkeys. AB - To examine the role of competitive and cooperative neural interactions within the intermediate layer of superior colliculus (SC), we elevated the basal SC neuronal activity by locally injecting a cholinergic agonist nicotine and analyzed its effects on saccade performance. After microinjection, spontaneous saccades were directed toward the movement field of neurons at the injection site (affected area). For visually guided saccades, reaction times were decreased when targets were presented close to the affected area. However, when visual targets were presented remote from the affected area, reaction times were not increased regardless of the rostrocaudal level of the injection sites. The endpoints of visually guided saccades were biased toward the affected area when targets were presented close to the affected area. After this endpoint effect diminished, the trajectories of visually guided saccades remained modestly curved toward the affected area. Compared with the effects on endpoints, the effects on reaction times were more localized to the targets close to the affected area. These results are consistent with a model that saccades are triggered by the activities of neurons within a restricted region, and the endpoints and trajectories of the saccades are determined by the widespread population activity in the SC. However, because increased reaction times were not observed for saccades toward targets remote from the affected area, inhibitory interactions in the SC may not be strong enough to shape the spatial distribution of the low-frequency preparatory activities in the SC. PMID- 15342716 TI - Tone frequency maps and receptive fields in the developing chinchilla auditory cortex. AB - Single-unit responses to tone pip stimuli were isolated from numerous microelectrode penetrations of auditory cortex (under ketamine anesthesia) in the developing chinchilla (laniger), a precocious mammal. Results are reported at postnatal day 3 (P3), P15, and P30, and from adult animals. Hearing sensitivity and spike firing rates were mature in the youngest group. The topographic representation of sound frequency (tonotopic map) in primary and secondary auditory cortex was also well ordered and sharply tuned by P3. The spectral temporal complexity of cortical receptive fields, on the other hand, increased progressively (past P30) to adulthood. The (purported) refinement of initially diffuse tonotopic projections to cortex thus seems to occur in utero in the chinchilla, where external (and maternal) sounds are considerably attenuated and might not contribute to the mechanism(s) involved. This compares well with recent studies of vision, suggesting that the refinement of the retinotopic map does not require external light, but rather waves of (correlated) spontaneous activity on the retina. In contrast, it is most probable that selectivity for more complex sound features, such as frequency stacks and glides, develops under the influence of the postnatal acoustic environment and that inadequate sound stimulation in early development (e.g., due to chronic middle ear disease) impairs the formation of the requisite intracortical (and/or subcortical) circuitry. PMID- 15342717 TI - Kinematic and kinetic constraints on arm, trunk, and leg segments in target reaching movements. AB - We studied target reaching tasks involving not only the arms but also the trunk and legs, which necessitated some trunk flexion. Such tasks can be successfully completed using an infinite number of combinations of segment motions due to the inherent kinematic redundancy with the excessive degrees of freedom (DOFs). Sagittal plane motions of six segments (shank, thigh, pelvis, trunk, humerus, and forearm) and dynamic torques of six joints (ankle, knee, hip, lumbar, shoulder, and elbow) were analyzed separately by principal component (PC) analyses to determine if there was a commonality among the shapes of the respective waveforms. Additionally, PC analyses were used to probe for constraining relationships among the 1) relative magnitudes of segment excursions and 2) the peak-to-peak dynamic joint torques. In summary, at the kinematic level, the tasks are simplified by the use of a single common waveform for all segment excursions with 89.9% variance accounted for (VAF), but with less fixed relationships among the relative scaling of the magnitude of segment excursions (62.2% VAF). However, at the kinetic level, the time course of the dynamic joint torques are not well captured by a single waveform (72.7% VAF), but the tasks are simplified by relatively fixed relationships among the scaling of dynamic joint torque magnitudes across task conditions (94.7% VAF). Taken together, these results indicate that, while the effective DOFs in a multi-joint task are reduced differently at the kinematic and kinetic levels, they both contribute to simplifying the neural control of these tasks. PMID- 15342718 TI - Cross-correlation and joint spectro-temporal receptive field properties in auditory cortex. AB - Recordings were made from the right primary auditory cortex in 17 adult cats using two eight-electrode arrays. We recorded the neural activity under spontaneous firing conditions and during random, multi-frequency stimulation, at 65 dB SPL, from the same units. Multiple single-unit (MSU) recordings (281) were stationary through 900 s of silence and during 900 s of stimulation. The cross correlograms of 545 MSU pairs with peak lag times within 10 ms from zero lag time were analyzed. Stimulation reduced the correlation in background activity, and as a result, the signal-to-noise ratio of correlated activity in response to the stimulus was enhanced. Reconstructed spectro-temporal receptive fields (STRFs) for coincident spikes showed larger STRF overlaps, suggesting that coincident neural activity serves to sharpen the resolution in the spectro-temporal domain. The cross-correlation for spikes contributing to the STRF depended much stronger on the STRF overlap than the cross-correlation during either silence or for spikes that did not contribute to the STRF (OUT-STRF). Compared with that for firings during silence, the cross-correlation for the OUT-STRF spikes was much reduced despite the unchanged firing rate. This suggests that stimulation breaks up the large neural assembly that exists during long periods of silence into a stimulus related one and maybe several others. As a result, the OUT-STRF spikes of the unit pairs, now likely distributed across several assemblies, are less correlated than during long periods of silence. Thus the ongoing network activity is significantly different from that during stimulation and changes afterng arousal during stimulation. PMID- 15342719 TI - Cortical injury affects short-term plasticity of evoked excitatory synaptic currents. AB - The hypothesis that plastic changes in the efficacy of excitatory neurotransmission occur in areas of chronic cortical injury was tested by assessing short-term plasticity of evoked excitatory synaptic currents (EPSCs) in neurons of partially isolated neocortical islands (undercut cortex). Whole cell recordings were obtained from layer V pyramidal neurons of sensorimotor cortical slices prepared from P36-P43 control and undercut rats. AMPA/kainate receptor mediated EPSCs elicited by stimuli delivered at 40 to 66.7 Hz exhibited more paired-pulse depression (PPD) in undercut cortex than control, the time constant of depression evoked by trains of 20- to 66.7-Hz stimuli was faster, and the steady-state amplitude of EPSCs reached after five to seven EPSCs was lower. An antagonist of the glutamate autoreceptor, group II mGluR, increased the steady state amplitude of EPSCs from undercut but not control cortex, suggesting that activation of presynaptic receptors by released glutamate is more prominent in undercut cortex. In contrast, the GABA(B) receptor antagonist (2S)-3-[[(1S)-1 (3,4-dichlorophenyl)ethyl]amino-2-hydroxypropyl](phenylmethyl)phosphinic acid had no effect. Increasing [Ca(2+)](o) from 2 to 4 mM increased PPD, with a smaller effect in neurons of the undercut. The I-V relationship of AMPA/kainate receptor mediated EPSCs was close to linear in both control and undercut neurons, and spermine had no significant effect on the EPSCs, suggesting that decreases in postsynaptic glutamate receptors containing the GluR2 subunit were not involved in the alterations in short-term plasticity. Results are compatible with an increase in the probability of transmitter release at excitatory synapses in undercut cortex due to functional changes in presynaptic terminals. PMID- 15342720 TI - A limited set of muscle synergies for force control during a postural task. AB - Recently developed computational techniques have been used to reduce muscle activation patterns of high complexity to a simple synergy organization and to bring new insights to the long-standing degrees of freedom problem in motor control. We used a nonnegative factorization approach to identify muscle synergies during postural responses in the cat and to examine the functional significance of such synergies for natural behaviors. We hypothesized that the simplification of neural control afforded by muscle synergies must be matched by a similar reduction in degrees of freedom at the biomechanical level. Electromyographic data were recorded from 8-15 hindlimb muscles of cats exposed to 16 directions of support surface translation. Results showed that as few as four synergies could account for >95% of the automatic postural response across all muscles and all directions. Each synergy was activated for a specific set of perturbation directions, and moreover, each was correlated with a unique vector of endpoint force under the limb. We suggest that, within the context of active balance control, postural synergies reflect a neural command signal that specifies endpoint force of a limb. PMID- 15342721 TI - Modulation of grasping forces during object transport. AB - Subjects held an instrumented object in a tripod grasp and moved it in the horizontal plane in various directions. The contact forces at the digits were measured and the grip force was decomposed into 2 components: a manipulating force responsible for accelerating the object and a grasping force responsible for holding the object steady. The grasping forces increased during the movement, reaching a peak near the time of peak velocity. The grasping forces also exhibited directional tuning, but this tuning was idiosyncratic for each subject. Although the overall grip forces should be modulated with acceleration, the load force did not vary during the task. Therefore the increase in the grasping force is not required to prevent slip. Rather, it is suggested that grasping force increases during translational motion to stabilize the orientation of grasped objects. PMID- 15342722 TI - Role of mossy fiber sprouting and mossy cell loss in hyperexcitability: a network model of the dentate gyrus incorporating cell types and axonal topography. AB - Mossy cell loss and mossy fiber sprouting are two characteristic consequences of repeated seizures and head trauma. However, their precise contributions to the hyperexcitable state are not well understood. Because it is difficult, and frequently impossible, to independently examine using experimental techniques whether it is the loss of mossy cells or the sprouting of mossy fibers that leads to dentate hyperexcitability, we built a biophysically realistic and anatomically representative computational model of the dentate gyrus to examine this question. The 527-cell model, containing granule, mossy, basket, and hilar cells with axonal projections to the perforant-path termination zone, showed that even weak mossy fiber sprouting (10-15% of the strong sprouting observed in the pilocarpine model of epilepsy) resulted in the spread of seizure-like activity to the adjacent model hippocampal laminae after focal stimulation of the perforant path. The simulations also indicated that the spatially restricted, lamellar distribution of the sprouted mossy fiber contacts reported in in vivo studies was an important factor in sustaining seizure-like activity in the network. In contrast to the robust hyperexcitability-inducing effects of mossy fiber sprouting, removal of mossy cells resulted in decreased granule cell responses to perforant-path activation in agreement with recent experimental data. These results indicate the crucial role of mossy fiber sprouting even in situations where there is only relatively weak mossy fiber sprouting as is the case after moderate concussive experimental head injury. PMID- 15342723 TI - Modulation of gaze-evoked blinks depends primarily on extraretinal factors. AB - Gaze-evoked blinks are contractions of the orbicularis oculi (OO)-the lid closing muscle-occurring during rapid movements of the head and eyes and result from a common drive to the gaze and blink motor systems. However, blinks occurring during shifts of gaze are often suppressed when the gaze shift is made to an important visual stimulus, suggesting that the visual system can modulate the occurrence of these blinks. In head-stabilized, human subjects, we tested the hypothesis that the presence of a visual stimulus was sufficient, but not necessary, to modulate OO EMG (OOemg) activity during saccadic eye movements. Rapid, reorienting movements of the eyes (saccades) were made to visual targets that remained illuminated (visually guided trials) or were briefly flashed (memory-guided trials) at different amplitudes along the horizontal meridian. We measured OOemg activity and found that the magnitude and probability of OOemg activity occurrence was reduced when a saccade was made to the memory of the spatial location as well as to the actual visual stimulus. The reduced OOemg activity occurred only when the location of the target was previously cued. OOemg activity occurred reliably with spontaneous saccades that were made to locations with no explicit visual stimulus, generally, back to the fixation location. Thus the modulation of gaze-evoked OOemg activity does not depend on the presence of visual information per se, but rather, results from an extraretinal signal. PMID- 15342724 TI - Visuotopic mapping through a multichannel stimulating implant in primate V1. AB - We report on our efforts to establish an animal model for the development and testing of a cortical visual prostheses. One-hundred-fifty-two electrodes were implanted in the primary visual cortex of a rhesus monkey. The electrodes were made from iridium with an activated iridium oxide film, which has a large charge capacity for a given surface area, and insulated with parylene-C. One-hundred fourteen electrodes were functional after implantation. The activity of small (2 3) neuronal clusters was first recorded to map the visually responsive region corresponding to each electrode. The animal was then trained in a memory (delayed) saccade task, first with a visual target, then to a target defined by direct cortical stimulation with coordinates specified by the stimulating electrode's mapped receptive field. The SD of saccade endpoints was approximately 2.5 larger for electrically stimulated versus visual saccades; nevertheless, when trial-to-trial scatter was averaged out, the correlation between saccade end points and receptive field locations was highly significant and approached unity after several months of training. Five electrodes were left unused until the monkey was fully trained; when these were introduced, the receptive field-saccade correlations were high on the first day of use (R = 0.85, P = 0.03 for angle, R = 0.98, P < 0.001 for eccentricity), indicating that the monkey had not learned to perform the task empirically by memorizing reward zones. The results of this experiment suggest the potential for rigorous behavioral testing of cortical visual prostheses in the macaque. PMID- 15342725 TI - The expression of matrix metalloproteinase-12 by oligodendrocytes regulates their maturation and morphological differentiation. AB - Matrix metalloproteinases (MMPs) have important roles in many processes of the developing CNS requiring proteolytic activity such as the migration of neuronal precursors, axonal outgrowth, and vascularization. Another developmental event involving proteolysis is myelin formation, whereby the extensive processes elaborated from oligodendrocytes (OLs) enwrap axons. Here we find MMP-12 transcripts to be produced by OLs in much higher levels than other MMP members examined. MMP-12 activity correlated with the ability of OLs to extend processes in vitro, suggesting a role for MMP-12 in the morphological differentiation of OLs. This was corroborated by results that OL lineage cells from MMP-12 null mice were retarded in their ability to differentiate morphologically and that this deficiency was overcome by the exogenous addition of active MMP-12. Finally, the maturation of oligodendrocyte precursor cells (OPCs) to OLs was significantly reduced in cultures from MMP-12 null mice compared with wild-type controls. We conclude that OL lineage cells express MMP-12 during their maturation and that MMP-12 activity has functional involvement both in maturation of OPCs and in the ability of OPCs and OLs to extend processes. PMID- 15342726 TI - Suprachiasmatic GABAergic inputs to the paraventricular nucleus control plasma glucose concentrations in the rat via sympathetic innervation of the liver. AB - Daily peak plasma glucose concentrations are attained shortly before awakening. Previous experiments indicated an important role for the biological clock, located in the suprachiasmatic nuclei (SCN), in the genesis of this anticipatory rise in plasma glucose concentrations by controlling hepatic glucose production. Here, we show that stimulation of NMDA receptors, or blockade of GABA receptors in the paraventricular nucleus of the hypothalamus (PVN) of conscious rats, caused a pronounced increase in plasma glucose concentrations. The local administration of TTX in brain areas afferent to the PVN revealed that an important part of the inhibitory inputs to the PVN was derived from the SCN. Using a transneuronal viral-tracing technique, we showed that the SCN is connected to the liver via both branches of the autonomic nervous system (ANS). The combination of a blockade of GABA receptors in the PVN with selective removal of either the sympathetic or parasympathetic branch of the hepatic ANS innervation showed that hyperglycemia produced by PVN stimulation was primarily attributable to an activation of the sympathetic input to the liver. We propose that the daily rise in plasma glucose concentrations is caused by an SCN-mediated withdrawal of GABAergic inputs to sympathetic preautonomic neurons in the PVN, resulting in an increased hepatic glucose production. The remarkable resemblance of the presently proposed control mechanism to that described previously for the control of daily melatonin rhythm suggests that the GABAergic control of sympathetic preautonomic neurons in the PVN is an important pathway for the SCN to control peripheral physiology. PMID- 15342727 TI - Glyceraldehyde-3-phosphate dehydrogenase is a GABAA receptor kinase linking glycolysis to neuronal inhibition. AB - Protein phosphorylation is crucial for regulating synaptic transmission. We describe a novel mechanism for the phosphorylation of the GABA(A) receptor, which mediates fast inhibition in the brain. A protein copurified and coimmunoprecipitated with the phosphorylated receptor alpha1 subunit; this receptor-associated protein was identified by purification and microsequencing as the key glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Molecular constructs demonstrated that GAPDH directly phosphorylates the long intracellular loop of GABA(A) receptor alpha1 subunit at identified serine and threonine residues. GAPDH and the alpha1 subunit were found to be colocalized at the neuronal plasma membrane. In keeping with the GAPDH/GABA(A) receptor molecular association, glycolytic ATP produced locally at plasma membranes was consumed for this alpha1 subunit phosphorylation, possibly within a single macrocomplex. The membrane-attached GAPDH is thus a dual-purpose enzyme, a glycolytic dehydrogenase, and a receptor-associated kinase. In acutely dissociated cortical neurons, the rundown of the GABA(A) responses was essentially attributable to a Mg(2+)-dependent phosphatase activity, which was sensitive to vanadate but insensitive to okadaic acid or fluoride. Rundown was significantly reduced by the addition of GAPDH or its reduced cofactor NADH and nearly abolished by the addition of its substrate glyceraldehyde-3-phosphate (G3P). The prevention of rundown by G3P was abolished by iodoacetamide, an inhibitor of the dehydrogenase activity of GAPDH, indicating that the GABA(A) responses are maintained by a glycolysis-dependent phosphorylation. Our results provide a molecular mechanism for the direct involvement of glycolysis in neurotransmission. PMID- 15342728 TI - GABA release and uptake regulate neuronal precursor migration in the postnatal subventricular zone. AB - In the postnatal subventricular zone (SVZ), astrocyte-like cells tightly encapsulate chains of migrating neuronal precursors, although an influence of the astrocyte-like cells on precursor migration has not yet been demonstrated. Cell migration was studied in acute sagittal brain slices to determine whether GABA signaling between astrocyte-like cells and neuronal precursors controls the speed of neuronal precursor migration in the anterior SVZ and rostral migratory stream of juvenile and adult mice. Application of GABA at 10 microm, a nondesensitizing concentration for GABA(A) receptors (GABA(A)Rs), reduced the rate (mean of approximately 50 microm/hr) of cell migration by 21% via GABA(A)R activation. Application of the GABA(A)R antagonist bicuculline enhanced the migration rate by 30%, suggesting that endogenous GABA tonically reduces the speed of cell migration via GABA(A)R activation. Using immunohistochemistry, we found that astrocyte-like cells express the high-affinity GABA transporter subtype GAT4 on processes ensheathing neuronal precursors that contain GABA. Inhibition of GABA uptake into astrocyte-like cells or enhancement of GABA release from neuronal precursors during high K(+) application further reduced the migration rate by increasing ambient GABA levels. GABA altered the migration speed by interfering with intracellular Ca(2+) signaling independently of cell depolarization, because high K(+) application did not alter the speed of cell migration in the presence of bicuculline. These data indicate that astrocyte-like cells create a microenvironment in which their uniquely positioned GABA transporters control the degree of GABA(A)R activation and the migration of neuronal precursors. PMID- 15342729 TI - Severe defects in dorsal thalamic development in low-density lipoprotein receptor related protein-6 mutants. AB - Mice with mutations in the Wnt coreceptor low-density lipoprotein receptor related protein-6 (LRP6) have a smaller and severely disorganized dorsal thalamus and lack thalamocortical projections. Using molecular markers, we showed that most dorsal thalamic and epithalamic neurons were missing, and most of the major dorsal thalamic nuclei were not identifiable. However, the ventral thalamus was essentially unaffected, although the dorsal thalamic defect leads to rostral displacement of portions of the ventral thalamus. Analysis of younger embryos showed that epithalamic and dorsal thalamic neurons were not produced at early stages of development, whereas ventral thalamic neurons were still produced. These defects were accompanied by improper formation of the boundary between dorsal and ventral thalamus, the zona limitans interthalamica (ZLI). Furthermore, the expression of an early marker of posterior forebrain development that marks the compartment from the midbrain-hindbrain junction to the ZLI (including the future dorsal thalamus, pretectum, and midbrain) was disrupted, supporting the idea that diencephalic development is abnormal from very early in embryogenesis. This study provides compelling in vivo evidence that thalamic development requires normal activity of the LRP6-mediated canonical Wnt signaling pathway. PMID- 15342730 TI - The isoazimuthal perception of sounds across distance: a preliminary investigation into the location of the audio egocenter. AB - Evidence indicates that both visual and auditory input may be represented in multiple frames of reference at different processing stages in the nervous system. Most models, however, have assumed that unimodal auditory input is first encoded in a head-centered reference frame. The present work tested this conjecture by measuring the subjective auditory egocenter in six blindfolded listeners who were asked to match the perceived azimuths of sounds that were alternately played between a surrounding arc of far-field speakers and a hand held point source located three different distances from the head. If unimodal auditory representation is head centered, then "isoazimuth" lines fitted to the matching estimates across distance should intersect near the midpoint of the interaural axis. For frontomedially arranged speakers, isoazimuth lines instead converged in front of the interaural axis for all listeners, often at a point between the two eyes. As far-field sources moved outside the visual field, however, the auditory egocenter location implied by the intersection of the isoazimuth lines retreated toward or even behind the interaural axis. Physiological and behavioral evidence is used to explain this change from an eye centered to a head-centered auditory egocenter as a function of source laterality. PMID- 15342731 TI - Reduction in size of perforated postsynaptic densities in hippocampal axospinous synapses and age-related spatial learning impairments. AB - A central problem in the neurobiology of normal aging is why learning is preserved in some aged individuals yet impaired in others. To investigate this issue, we examined whether age-related deficits in spatial learning are associated with a reduction in postsynaptic density (PSD) area in hippocampal excitatory synapses (i.e., with a structural modification that is likely to have a deleterious effect on synaptic function). A hippocampus-dependent version of the Morris water maze task was used to separate Long-Evans male rats into young adult, aged learning-unimpaired, and equally aged learning-impaired groups. Axospinous synapses from the CA1 stratum radiatum were analyzed using systematic random sampling and serial section analyses. We report that aged learning impaired rats exhibit a marked ( approximately 30%) and significant reduction in PSD area, whereas aged learning-unimpaired rats do not. The observed structural alteration involves a substantial proportion of perforated synapses but is not observed in nonperforated synapses. These findings support the notion that many hippocampal perforated synapses become less efficient in aged learning-impaired rats, which may contribute to cognitive decline during normal aging. PMID- 15342732 TI - Selective neurotoxic lesions of basolateral and central nuclei of the amygdala produce differential effects on fear conditioning. AB - In the fear conditioning literature, it is generally hypothesized that neurons in the basolateral amygdalar complex (BLA) (lateral and basal nuclei) support the formation of conditioned fear memory and project to neurons in the central nucleus (CeA) for the expression of conditioned fear responses. According to this serial processing-transmission view, damage to either BLA or CeA would comparably disrupt the expression of a variety of conditioned fear responses. In the present study, we further investigated the roles of BLA and CeA in fear conditioning by concurrently assessing freezing and 22 kHz ultrasonic vocalization (USV) as dependent measures of fear in rats. Selective neurotoxins, NMDA for the BLA and ibotenic acid for the CeA, were used to destroy intrinsic neurons [evidenced by thionin dye and NeuN (neuronal nuclei) antibody stainings] without damaging the fibers of passage (confirmed by myelin staining). During the 10 tone-footshock paired training, postshock freezing and USV responses were significantly impaired in BLA-lesioned animals, whereas CeA-lesioned animals exhibited only mild deficits. Similarly, conditioned fear responses assessed 24 hr after training were severely reduced in BLA-lesioned animals but not in CeA-lesioned animals. In contrast to ibotenic lesions of the CeA, small electrolytic lesions of the CeA strongly affected both postshock and conditioned freezing and USV. Together, these results do not support the currently espoused BLA-to-CeA serial processing transmission view of fear conditioning. Instead, the expression of conditioned fear appears to primarily involve BLA projections that course through the CeA en route to downstream fear response structures. PMID- 15342733 TI - An element in the alpha1-tubulin promoter is necessary for retinal expression during optic nerve regeneration but not after eye injury in the adult zebrafish. AB - We have shown previously that a 1.696 kb upstream fragment of the goldfish alpha1 tubulin promoter was capable of driving green fluorescent protein (GFP) expression in the developing and regenerating zebrafish CNS in a pattern closely mimicking the endogenous alpha1-tubulin gene. Comparison of fish and rat alpha1 tubulin promoters identified a 64 bp region with a conserved repetitive homeodomain (HD) consensus sequence core (TAAT) and a nearby basic helix-loop helix binding E-box sequence (CANNTG), which led us to speculate that it could be of importance for regulating alpha1-tubulin gene transcription. To address this issue, we examined the ability of deletion mutants of the 1.696 kb promoter to drive expression of GFP in zebrafish retinal cells under normal conditions and after injury. Interestingly, although wild-type 1.696 kb and mutant promoters, lacking the E-box and/or HD sequences, exhibited rather similar patterns of GFP expression in the developing retina, significant differences were noticed in the mature retina. First, although the 1.696 kb promoter directed transgene expression to retinal neurons and progenitor cells, the activity of mutant promoters was drastically reduced. Second, we found that the E-box and HD sequences were necessary for transgene reinduction during optic nerve regeneration, but were not as important for transgene expression in regenerating retinal neurons after eye injury. In this latter lesion model, remarkably, both 1.696 kb and mutant promoters targeted GFP expression to Muller glia-like cells, some of which re-entered the cell cycle. These new findings will be useful for identifying the molecular signals necessary for successful CNS regeneration. PMID- 15342735 TI - Hippocampal plasticity across multiple days of exposure to novel environments. AB - The hippocampus is essential for learning complex spatial relationships, but little is known about how hippocampal neural activity changes as animals learn about a novel environment. We studied the formation of new place representations in rats by examining the changes in place-specific firing of neurons in the CA1 region of the hippocampus and the relationship between these changes and behavioral change across multiple days of exposure to novel places. We found that many neurons showed very rapid changes on the first day of exposure to the novel place, including many cases in which a previously silent neuron developed a place field over the course of a single pass through the environment. Across the population, the largest changes in neural activity occurred on day 2 of exposure to a novel place, but only if the animal had little experience (<4 min) in that location on day 1. Longer exposures on day 1 were associated with smaller changes on day 2, suggesting that hippocampal neurons required 5-6 min of experience to form a stable spatial representation. Even after the representation stabilized, the animals' behavior remained different in the novel places, suggesting that other brain regions continued to distinguish novel from familiar locations. These results show that the hippocampus can form new spatial representations quickly but that stable hippocampal representations are not sufficient for a place to be treated as familiar. PMID- 15342734 TI - Umami taste responses are mediated by alpha-transducin and alpha-gustducin. AB - The sense of taste comprises at least five distinct qualities: sweet, bitter, sour, salty, and umami, the taste of glutamate. For bitter, sweet, and umami compounds, taste signaling is initiated by binding of tastants to G-protein coupled receptors in specialized epithelial cells located in the taste buds, leading to the activation of signal transduction cascades. Alpha-gustducin, a taste cell-expressed G-protein alpha subunit closely related to the alpha transducins, is a key mediator of sweet and bitter tastes. Alpha-gustducin knock out (KO) mice have greatly diminished, but not entirely abolished, responses to many bitter and sweet compounds. We set out to determine whether alpha-gustducin also mediates umami taste and whether rod alpha-transducin (alpha(t-rod)), which is also expressed in taste receptor cells, plays a role in any of the taste responses that remain in alpha-gustducin KO mice. Behavioral tests and taste nerve recordings of single and double KO mice lacking alpha-gustducin and/or alpha(t-rod) confirmed the involvement of alpha-gustducin in bitter (quinine and denatonium) and sweet (sucrose and SC45647) taste and demonstrated the involvement of alpha-gustducin in umami [monosodium glutamate (MSG), monopotassium glutamate (MPG), and inosine monophosphate (IMP)] taste as well. We found that alpha(t-rod) played no role in taste responses to the salty, bitter, and sweet compounds tested or to IMP but was involved in the umami taste of MSG and MPG. Umami detection involving alpha-gustducin and alpha(t-rod) occurs in anteriorly placed taste buds, however taste cells at the back of the tongue respond to umami compounds independently of these two G-protein subunits. PMID- 15342736 TI - Visual neurons in the pigeon brain encode the acceleration of stimulus motion. AB - Seeing target motion is a vital capability of the visual system in humans and animals. Physically, motion is described by its acceleration, speed, and direction. Motion-sensitive neurons in all the visual areas examined to date are selective for the direction and speed of motion. Here, we show by single-unit recording that one-third of motion-sensitive neurons in the pigeon's pretectal nucleus also encode the acceleration of stimulus motion. These neurons are characterized by plateau-shaped speed-tuning curves in which the firing rate is the same over a wide range of speeds, a feature that allows these neurons to encode unambiguously the rate of change of speed over time. Acceleration sensitive neurons also show transient responses to the offset of motion in the preferred and/or nonpreferred directions; acceleration-insensitive neurons do not. We observed the same sensitivity to target acceleration for brief ramps of stimulus speed and for sinusoidal modulation of speed. The locations of acceleration-sensitive and -insensitive neurons are segregated in the pretectal nucleus. The visual responses of pretectal neurons indicate that the visual and vestibular systems share not only a spatial but also a temporal reference frame that can detect the acceleration produced by self-motion of an organism. PMID- 15342737 TI - Chronic morphine treatment reduces recovery from opioid desensitization. AB - Tolerance and dependence result from long-term exposure to opioids, and there is growing evidence linking acute receptor desensitization to these more long-term processes. Receptor desensitization encompasses a series of events leading to the loss of receptor function and internalization. This study examines the onset and recovery from desensitization in locus ceruleus neurons recorded in brain slices taken from animals that have been chronically treated with morphine. After chronic morphine treatment, desensitization was altered as follows. First, the rate of desensitization was increased. Second, recovery from desensitization was always incomplete, even after a brief (1-2 min) exposure to agonist. This contrasts with experiments in controls in which recovery from desensitization, after a brief exposure to agonist, was complete within 25 min. Finally, morphine 6-beta-D-glucuronide, a metabolite of morphine that was ineffective at causing desensitization in controls, induced significant desensitization in slices from morphine-treated animals. When brain slices from controls were treated with inhibitors of PKC or monensin, agents known to compromise G-protein-coupled receptor resensitization, desensitization was increased, and recovery was significantly reduced. These results indicate that receptor resensitization maintains signaling during periods of intense and sustained stimulation. After chronic morphine treatment, desensitization is potentiated, and receptor resensitization is compromised. PMID- 15342738 TI - Neutralization of transthyretin reverses the neuroprotective effects of secreted amyloid precursor protein (APP) in APPSW mice resulting in tau phosphorylation and loss of hippocampal neurons: support for the amyloid hypothesis. AB - Alzheimer's disease (AD) may be caused by the abnormal processing of the amyloid precursor protein (APP) and the accumulation of beta-amyloid (Abeta). The amyloid precursor protein can be proteolytically cleaved into multiple fragments, many of which have distinct biological actions. Although a high level of Abeta can be toxic, the alpha-secretase cleaved APP (sAPPalpha) is neuroprotective. However, the mechanism of sAPPalpha protection is unknown. Here, we show that sAPPalpha increases the expression levels of several neuroprotective genes and protects organotypic hippocampal cultures from Abeta-induced tau phosphorylation and neuronal death. Antibody interference and small interfering RNA knock-down demonstrate that the sAPPalpha-driven expression of transthyretin and insulin like growth factor 2 is necessary for protection against Abeta-induced neuronal death. Mice overexpressing mutant APP possess high levels of sAPPalpha and transthyretin and do not develop the tau phosphorylation or neuronal loss characteristic of human AD. Chronic infusion of an antibody against transthyretin into the hippocampus of mice overexpressing APP with the Swedish mutation (APP(Sw)) leads to increased Abeta, tau phosphorylation, and neuronal loss and apoptosis within the CA1 neuronal field. Therefore, the elevated expression of transthyretin is mediated by sAPPalpha and protects APP(Sw) mice from developing many of the neuropathologies observed in AD. PMID- 15342739 TI - Muscarinic receptor modulation of slow afterhyperpolarization and phasic firing in rat supraoptic nucleus neurons. AB - A slow posttrain afterhyperpolarization (sAHP) was studied in rat magnocellular neurosecretory cells (MNCs) in vitro. The sAHP was isolated from other afterpotentials by blocking the depolarizing afterpotential (DAP) with Cs(+) and the medium afterhyperpolarization (mAHP) with apamin. The sAHP amplitude increased logarithmically with activity ( approximately 3 mV per e-fold increase in number of impulses) and, when firing stopped, decayed exponentially with a time constant of 2 sec. The sAHP was associated with increased membrane conductance, and its amplitude varied linearly with voltage, reversing at the K(+) equilibrium potential. The sAHP was blocked by Cd(2+) but not by charybdotoxin or iberiotoxin, blockers of intermediate- and big-conductance-type Ca(2+)-dependent K(+) (K(Ca)) channels. The sAHP was reversibly inhibited by muscarine, an effect antagonized by atropine, indicating involvement of muscarinic cholinergic receptors. Muscarine did not affect Ca(2+)-dependent features of action potentials, DAPs, or the mAHP in MNCs, indicating selective modulation of K(Ca) channels causing the sAHP. Muscarinic inhibition of the sAHP enhanced plateau potentials and increased the mean firing rate and duration of afterdischarges that followed spike trains evoked from voltages near threshold. Similarly, the frequency and duration of the spontaneous phasic bursts that characterize physiologically activated vasopressin-releasing MNCs were enhanced by muscarine. MNCs thus express apamin- and voltage-insensitive K(Ca) channels that mediate an sAHP. The activity dependence and kinetics of the sAHP cause it to mask DAPs in a manner that attenuates the amplitude of plateau potentials. Muscarinic inhibition of the sAHP provides an effective mechanism for promoting phasic firing in MNCs. PMID- 15342740 TI - Brain-derived neurotrophic factor regulates the onset and severity of motor dysfunction associated with enkephalinergic neuronal degeneration in Huntington's disease. AB - The mechanism that controls the selective vulnerability of striatal neurons in Huntington's disease is unclear. Brain-derived neurotrophic factor (BDNF) protects striatal neurons and is regulated by Huntingtin through the interaction with the neuron-restrictive silencer factor. Here, we demonstrate that the downregulation of BDNF by mutant Huntingtin depends on the length and levels of expression of the CAG repeats in cell cultures. To analyze the functional effects of these changes in BDNF in Huntington's disease, we disrupted the expression of bdnf in a transgenic mouse model by cross-mating bdnf(+/ -) mice with R6/1 mice. Thus, we compared transgenic mice for mutant Huntingtin with different levels of BDNF. Using this double mutant mouse line, we show that the deficit of endogenous BDNF modulates the pathology of Huntington's disease. The decreased levels of this neurotrophin advance the onset of motor dysfunctions and produce more severe uncoordinated movements. This behavioral pathology correlates with the loss of striatal dopamine and cAMP-regulated phosphoprotein-32-positive projection neurons. In particular, the insufficient levels of BDNF cause specific degeneration of the enkephalinergic striatal projection neurons, which are the most affected cells in Huntington's disease. This neuronal dysfunction can specifically be restored by administration of exogenous BDNF. Therefore, the decrease in BDNF levels plays a key role in the specific pathology observed in Huntington's disease by inducing dysfunction of striatal enkephalinergic neurons that produce severe motor dysfunctions. Hence, administration of exogenous BDNF may delay or stop illness progression. PMID- 15342741 TI - The type IV-specific phosphodiesterase inhibitor rolipram and its effect on hippocampal long-term potentiation and synaptic tagging. AB - We investigated the effects of rolipram, a selective cAMP phosphodiesterase (PDE) inhibitor, on late plastic events during functional CA1 plasticity in vitro in rat hippocampal slices. We present data showing that an early form of long-term potentiation (LTP) (early-LTP) that normally decays within 2-3 hr can be converted to a lasting LTP (late-LTP) if rolipram is applied during tetanization. This rolipram-reinforced LTP (RLTP) was NMDA receptor and protein synthesis dependent. cAMP formation in region CA1 during late-LTP requires dopaminergic receptor activity (Frey et al., 1989, 1990). Thus, we studied whether RLTP was influenced by inhibitors of the D(1)/D(5) receptor. Application of the specific D(1)/D(5) antagonist SCH23390 (0.1 microm) did not prevent RLTP, suggesting that the phosphodiesterase inhibitor acts downstream of the D(1)/D(5) receptors. We also studied whether rolipram can interact with processes of synaptic tagging, because RLTP was also dependent on protein synthesis, similar to late-LTP. Inhibition of PDE and subsequent induction of RLTP in one synaptic population were able to transform early-LTP into late-LTP in a second, independent synaptic population of the same neurons. This supports our hypothesis that cAMP-dependent processes are directly involved in the synthesis of plasticity-related proteins. PMID- 15342742 TI - Noradrenergic inputs mediate state dependence of auditory responses in the avian song system. AB - Norepinephrine (NE) plays a complex role in the behavioral state-dependent regulation of sensory processing. However, the role of forebrain NE action in modulating high-order sensory activity has not been directly addressed. In this study, we take advantage of the discrete, feedforward organization of the avian song system to identify a site and mechanism of NE action underlying state dependent modulation of sensory processing. We have developed an experimental paradigm in which brief arousal repeatedly suppresses song system auditory responsiveness. Using pharmacological manipulations in vivo, we show that infusion of alpha-adrenergic antagonists into the NIf (nucleus interfacialis of the nidopallium), an auditory forebrain area, blocks this state-dependent modulation. We also demonstrate dose-dependent enhancement and suppression of song system auditory response properties by NE and adrenergic agonists. Our results demonstrate that noradrenergic release in a single forebrain area is a mechanism underlying behavioral state-dependent regulation of auditory processing in a neural system specialized for vocal learning. PMID- 15342743 TI - Olfactory receptor proteins in axonal processes of chemosensory neurons. AB - Olfactory receptors are supposed to act not only as molecular sensors for odorants but also as cell recognition molecules guiding the axons of olfactory neurons to their appropriate glomerulus in the olfactory bulb. This concept implies that olfactory receptor proteins are located in sensory cilia and in the axons. To approach this critical issue, antibodies were generated against two peptides, one derived from olfactory receptor mOR256-17, one derived from the "mOR37" subfamily. By means of immunohistochemistry and double-labeling studies using transgenic mouse lines as well as Western blot analyses, it was demonstrated that the newly generated antibodies specifically recognized the receptor proteins. To scrutinize the hypothesis that olfactory receptor proteins may also be present in the axonal processes and the nerve terminals, serial sections through the olfactory bulb were probed with the antibodies. Two glomeruli in each bulb were stained by anti-mOR256-17, one positioned in the medial, one in the lateral hemisphere. Fiber bundles approaching the glomeruli through the outer nerve layer also displayed intense immunofluorescence. A similar picture emerged for the antibody anti-mOR37, a small number of glomeruli in the ventral domain of the bulb was stained. On serial sections through the olfactory bulb of mOR37-transgenic mouse lines, double-labeling experiments demonstrated that distinct immunoreactive glomeruli corresponded to glomeruli that were targeted by neurons expressing a particular member of the mOR37 receptor subfamily. These data indicate that olfactory receptor (OR) proteins are indeed present in the axonal processes and nerve terminals of olfactory sensory neurons, thus supporting the notion that ORs may participate in the molecular processes underlying the fasciculation and targeting of olfactory axons. PMID- 15342744 TI - Impaired repression at a vasopressin promoter polymorphism underlies overexpression of vasopressin in a rat model of trait anxiety. AB - Two inbred rat lines have been developed that show either high (HAB) or low (LAB) anxiety-related behavior. The behavioral phenotype correlates with arginine vasopressin (AVP) expression at the level of the hypothalamic paraventricular nucleus (PVN), but not supraoptic nucleus, with HAB animals overexpressing the neuropeptide in both magnocellular and parvocellular subdivisions of the PVN. We detected a number of single nucleotide polymorphisms (SNPs) in the AVP locus that differ between the HAB and LAB animals, two of which were embedded in cis regulatory elements. The HAB-specific allele of the AVP gene promoter occurs in 1.5% of outbred Wistar rats and is more transcriptionally active in vivo, as revealed by allele-specific transcription studies in cross-mated HAB/LAB F1 animals. Interestingly, one specific SNP [A(-1276)G] conferred reduced binding of the transcriptional repressor CArG binding factor A (CBF-A) in the HAB allele, the consequent differential regulation of the AVP promoter resulting in an overexpression of AVP in vitro and in vivo. Furthermore, CBF-A is highly coexpressed in AVP-containing neurons of the PVN supporting an important role for regulation of AVP gene expression in vivo. Taken together, our results demonstrate a role for an AVP gene polymorphism and CBF-A in elevated AVP expression in the PVN of HAB rats likely to contribute to their behavioral and neuroendocrine phenotype. PMID- 15342745 TI - Changing ethical standards in scientific publication. PMID- 15342746 TI - Potential donor rotational bone grafts using vascular territories in the foot and ankle. AB - BACKGROUND: An improved understanding of the extraosseous and intraosseous blood supply of the distal aspect of the tibia, distal aspect of the fibula, cuboid, and cuneiforms should identify vascular territories that would enable surgeons to perform rotational vascularized pedicle bone-grafting procedures in the foot and ankle. METHODS: We investigated the blood supply of twenty cadaveric lower extremities using two vascular injection techniques. In order to define the extraosseous and intraosseous arterial anatomy in this region, ten specimens were sequentially subjected to injection with Batson's compound, soft-tissue digestion, and bone-clearing according to a modified Spalteholz technique. To further characterize the extraosseous vascular anatomy, the other ten specimens were injected with latex and dissected. RESULTS: We identified a consistent and previously unnamed blood supply to the distal aspect of the tibia, distal aspect of the fibula, cuboid, and cuneiforms. Four vessels, each present in all of our specimens, provided distinct vascular territories to bone. A branch of the proximal lateral tarsal artery supplied a consistent vascular territory in the cuboid with an average of fifteen nutrient vessels. Similarly, a branch of the distal medial tarsal artery to the first cuneiform supplied an average of nine nutrient vessels superior to the tibialis anterior tendon insertion. A branch of the anterior lateral malleolar artery to the fibula supplied an average of seven nutrient vessels to the lateral malleolus. A branch of the distal lateral tarsal artery provided the midsection of the third cuneiform with an average of seven nutrient vessels. In the latex-injected specimens, harvesting of the vascularized pedicle bone grafts designed from these data demonstrated their anatomic plausibility and arcs of rotation. CONCLUSIONS: Four new rotational vascularized pedicle bone grafts have been identified in the foot and ankle. These grafts were present in all of our specimens, were well vascularized, had wide arcs of rotation, and were relatively easy to harvest. PMID- 15342747 TI - Fracture of the neck of the femur after surface arthroplasty of the hip. AB - BACKGROUND: There are two main modes of failure of the femur following surface arthroplasty of the hip: femoral neck fracture and aseptic loosening. The purpose of the present study was to present our experience with femoral neck fractures that occurred after metal-on-metal hybrid surface arthroplasty and to assess their cause. METHODS: A series of 600 metal-on-metal surface arthroplasties was performed between late 1996 and early 2003. Failures that occurred during this period were assessed radiographically and with implant retrieval analysis to determine their cause. RESULTS: Five femoral neck fractures occurred in this series (prevalence, 0.83%). Four of the five fractures occurred at the component neck junction within the first five months (average, three months) after surgery. All five fractures were associated with a traumatic episode, but all five also were associated with structural and/or technical risk factors, which we believe weakened the femoral neck. The most important technical deficiency that contributed to three of the five fractures was the failure to cover all of the reamed bone with the component. CONCLUSIONS: It is important to avoid or at least minimize notching of the femoral neck by performing the cylindrical reaming at the recommended angle of 140 degrees and to stop reaming before the reamer touches the lateral cortex. Osteophytes should be removed judiciously only if there is notable impingement when the hip is flexed to 90 degrees and internally rotated. We believe that understanding the factors that contribute to femoral neck fracture after surface arthroplasty may reduce the prevalence of this mode of failure. PMID- 15342748 TI - Replacement of the torn posterior cruciate ligament with a mid-third patellar tendon graft with use of a modified tibial inlay method. AB - BACKGROUND: The tibial inlay method for reconstruction of the posterior cruciate ligament has been performed with the patient in the prone or lateral decubitus position. The purpose of this report is to present a modification of this method wherein the patient is positioned supine throughout the procedure. METHODS: Between May 1995 and September 1998, twelve patients who had an isolated tear of the posterior cruciate ligament underwent reconstruction with use of the modified tibial inlay technique. Eleven patients were evaluated after a minimum duration of follow-up of two years. Stability was measured on posterior stress radiographs and with a maximum manual displacement test performed with a KT-1000 arthrometer. Clinical evaluation was carried out with use of the scoring systems of the Orthopadische Arbeitsgruppe Knie and the International Knee Documentation Committee. Second-look arthroscopy was performed in five patients at the time of follow-up. RESULTS: The mean side-to-side difference in displacement (and standard deviation) was reduced from 10.8 +/- 1.9 mm preoperatively to 3.4 +/- 2.4 mm at the time of follow-up as measured on the stress radiographs, and it was reduced from 9.0 +/- 2.1 mm preoperatively to 1.8 +/- 1.2 mm at the time of follow-up as measured with the KT-1000 arthrometer. The average Orthopadische Arbeitsgruppe Knie score was improved from 71.6 +/- 6.8 to 92.5 +/- 4.8 points. All eleven patients had a satisfactory clinical outcome at the time of the final clinical evaluation. The second-look arthroscopic examination in the five patients showed no evidence of partial tearing or abrasion of the graft. CONCLUSIONS: Use of our modified tibial inlay technique for reconstruction of the posterior cruciate ligament achieved a good clinical result in eleven of twelve patients. The advantages of the technique are (1) minimal tendon abrasion at the posterior opening of the tibial tunnel, and (2) elimination of the need to change the patient's position during surgery. PMID- 15342749 TI - Surgical management of trapezius palsy. AB - BACKGROUND: Injury to the spinal accessory nerve in the posterior cervical triangle leads to paralysis of the trapezius muscle. The aim of this study was to determine the indications for nerve repair or reconstructive surgery according to the etiology, the duration of the preoperative delay, and specific patient characteristics. METHODS: Of twenty-seven patients with a trapezius palsy, twenty were treated with neurolysis or surgical repair (direct or with a graft) of the spinal accessory nerve and seven were treated with the Eden-Lange muscle transfer procedure. Lymph node biopsy was the main cause of the nerve injury. The nerve repairs were performed at an average of seven months after the injury, and the reconstructive procedures were done at an average of twenty-eight months. Nerve repair was performed for iatrogenic injuries of the spinal accessory nerve, within twenty months after the onset of symptoms, and in one patient with spontaneous palsy. Reconstructive surgery was performed for cases of trapezius palsy secondary to radical neck dissection, for spontaneous palsies, and after failure of nerve repair or neurolysis. The mean follow-up period was thirty-five months. The functional outcome was assessed clinically on the basis of active shoulder abduction, pain, strength of the trapezius on manual muscle-testing, and level of subjective patient satisfaction. RESULTS: The results were good or excellent in sixteen of the twenty patients treated with nerve repair and in four of the seven patients treated with the Eden-Lange procedure. Poor results were seen in older patients and in patients with a previous radical neck dissection. CONCLUSIONS: Good results can be expected from a repair of the spinal accessory nerve if it is performed within twenty months after the injury, as the nerve is basically a purely motor nerve and the distance from the injury to the motor end plates is short. Muscle transfer should be performed in patients with spontaneous trapezius palsy, when previous nerve surgery has failed, or when the time from the injury to treatment is over twenty months. Treatment is less likely to succeed when the patient is older than fifty years of age or the palsy was due to a radical neck dissection, penetrating injury, or spontaneous palsy. PMID- 15342750 TI - Long-term follow-up of female patients with idiopathic scoliosis treated with the Wilmington orthosis. AB - BACKGROUND: A spinal orthosis is commonly utilized in the nonoperative treatment of idiopathic scoliosis. The purpose of this study was to evaluate the long-term radiographic and functional outcomes of female patients with idiopathic scoliosis who had completed a program of treatment with the Wilmington thoracic-lumbar spinal orthosis. METHODS: We retrospectively reviewed the clinical records and radiographs of all female patients who had successfully completed a course of treatment with the orthosis between 1973 and 1983. Ninety-one patients met the criteria for inclusion, and fifty-five women returned for a follow-up evaluation. Their mean age was thirty-one years at the time of follow-up, which was carried out at a mean of 14.6 years after the completion of treatment. The patients were evaluated clinically and radiographically, and they each completed a comprehensive questionnaire assessing their ability to perform twenty-six activities of daily living, their overall physical appearance, the cosmetic appearance of the back, their self-image, and the severity of any back pain. The questionnaire was also administered to a control group of fifty-five women without scoliosis matched for age, number of children, and occupation. RESULTS: Seven patients (13%) demonstrated >or=5 degrees of progression of the curve, compared with the curve at the start of treatment, after discontinuing use of the orthosis. No curve progressed >17 degrees compared with the deformity at the time of the initial treatment. There was no significant overall difference between the orthotic treatment group and the control group in terms of back pain, physical activities, functional activities (with the exception of shopping) or self-care activities. As a group, the patients reported significantly greater difficulty with selected positional activities (p = 0.007). Fifty-one (93%) of the fifty five treated women reported no subjective deterioration in their physical appearance, the cosmetic appearance of the back, or their self-image in the period since they discontinued using the brace. CONCLUSIONS: The majority of patients who successfully complete treatment with a Wilmington thoracic-lumbar spinal orthosis for idiopathic scoliosis with an initial magnitude of between 20 degrees and 45 degrees can anticipate that the curve will remain stable into middle adulthood. Any apparent correction of the curve that occurs during treatment can be expected to be lost over time, resulting in a deformity that is equal or nearly equal in magnitude to that measured at the initiation of the orthotic management. Because some patients did demonstrate some progression of the curve by the third or fourth decade of life, it is reasonable to recommend a spinal radiograph during that time to monitor the status of the curve. PMID- 15342751 TI - Loss of fixation of the volar lunate facet fragment in fractures of the distal part of the radius. AB - BACKGROUND: The purpose of the present study is to report on a cohort of patients with a volar shearing fracture of the distal end of the radius in whom the unique anatomy of the distal cortical rim of the radius led to failure of support of a volar ulnar lunate facet fracture fragment. METHODS: Seven patients with a volar shearing fracture of the distal part of the radius who lost fixation of a volar lunate facet fragment with subsequent carpal displacement after open reduction and internal fixation were evaluated at an average of twenty-four months after surgery. One fracture was classified as B3.2 and six were classified as B3.3 according to the AO comprehensive classification system. All seven fractures initially were deemed to have an adequate reduction and internal fixation. Four patients required repeat open reduction and internal fixation, and one underwent a radiocarpal arthrodesis. At the time of the final follow-up, all patients were assessed with regard to their self-reported level of functioning and with use of Sarmiento's modification of the system of Gartland and Werley. RESULTS: At a mean of two years after the injury, six patients had returned to their previous level of function. The result was considered to be excellent for one patient, good for four, and fair for two. The average wrist extension was 48 degrees, or 75% of that of the uninjured extremity. The average wrist flexion was 37 degrees, or 64% of that of the uninjured extremity. The one patient who underwent radiocarpal arthrodesis had achievement of a solid union. The four patients who underwent repeat internal fixation had maintenance of reduction of the lunate facet fragment. The two patients who declined additional operative intervention had persistent dislocation of the carpus with the volar lunate facet fragment. CONCLUSIONS: The stability of comminuted fractures of the distal part of the radius with volar fragmentation is determined not only by the reduction of the major fragments but also by the reduction of the small volar lunate fragment. The unique anatomy of this region may prevent standard fixation devices for distal radial fractures from supporting the entire volar surface effectively. It is preferable to recognize the complexity of the injury prior to the initial surgical intervention and to plan accordingly. PMID- 15342752 TI - Association between hospital and surgeon procedure volume and the outcomes of total knee replacement. AB - BACKGROUND: The annual volume of major cardiovascular and oncologic procedures performed in hospitals and by surgeons has been inversely associated with the rates of perioperative mortality and complications. The relationship between hospital and surgeon volume and perioperative outcomes following total knee replacement has received little study. METHODS: We analyzed claims data for Medicare patients who had elective primary total knee replacement between January 1 and August 31, 2000. Hospital and surgeon volumes were defined as the number of primary and revision total knee replacements performed in the hospital or by the surgeon in Medicare recipients in 2000. We examined the associations between the annual volumes of total knee replacement performed in the hospitals and by the surgeons and the rates of mortality and complications (infection, pulmonary embolus, myocardial infarction, or pneumonia) in the first ninety days postoperatively. The analyses were adjusted for age, gender, comorbid conditions, Medicaid eligibility (a marker of low income), and arthritis diagnosis. Analyses of hospital volume were adjusted for surgeon volume and vice versa. RESULTS: Twenty-five percent of the primary total knee replacements were done by surgeons who performed twelve of these procedures or fewer in the Medicare population annually, and 11% were done in hospitals with an annual volume of twenty-five of these procedures or fewer. Compared with the patients who had a primary total knee replacement in hospitals with an annual volume of twenty-five procedures or fewer, those managed in hospitals with an annual volume exceeding 200 procedures had a lower risk of pneumonia (odds ratio, 0.65; 99% confidence interval, 0.47 to 0.90) and any of the adverse outcomes examined (death, pneumonia, pulmonary embolus, acute myocardial infarction, or deep infection) (odds ratio, 0.74; 99% confidence interval, 0.60 to 0.90). Similarly, patients who had a primary total knee replacement done by surgeons who performed more than fifty such procedures in Medicare recipients annually had a lower risk of pneumonia (odds ratio, 0.72; 99% confidence interval, 0.54 to 0.95) and any adverse outcome (odds ratio, 0.81; 99% confidence interval, 0.68 to 0.98) compared with patients of surgeons with an annual volume of twelve procedures or fewer. CONCLUSIONS: Patients managed at hospitals and by surgeons with greater volumes of total knee replacement have lower risks of perioperative adverse events following primary total knee replacement. Patients and clinicians should incorporate these findings into discussions about selecting a surgeon and a hospital for total knee replacement. These data should also be integrated into the policy debate about the advantages and drawbacks of regionalizing total joint replacement to high-volume centers. PMID- 15342753 TI - Subchondral fatigue fracture of the femoral head in military recruits. AB - BACKGROUND: Subchondral stress fracture of the femoral head is a rare condition that usually occurs as an insufficiency fracture in people with poor bone quality. We evaluated the clinical characteristics of subchondral fatigue fractures of the femoral head that occurred in young, healthy military recruits. METHODS: Between January 1998 and November 2001, seven subchondral fatigue fractures of the femoral head were treated in five patients. The characteristics of this condition were ascertained by assessing the clinical course as well as radiographs, bone scintigrams, and magnetic resonance images. RESULTS: All patients were male military recruits in their early twenties in whom pain had developed within five months after recruitment. Definite abnormal findings were observed on the initial radiographs of four hips in three patients, and the femoral head was markedly collapsed in two of these four hips. Bone scintigrams were made of five hips in four patients, and all of them showed increased radionuclide uptake in the femoral head. In all affected hips, magnetic resonance images demonstrated a localized or diffuse bone-marrow-edema pattern in the femoral head and/or neck. A subchondral fracture line (a magnetic resonance crescent sign) was identified in all hips. In the patients who did not have collapse of the femoral head, the pain decreased gradually and disappeared completely within six months, with correspondingly improved findings on sequential magnetic resonance images. The patients with femoral head collapse were treated with total hip arthroplasty or an iliac bone strut graft. CONCLUSIONS: When a military recruit or an athlete reports hip pain, a diagnosis of subchondral fatigue fracture of the femoral head should be considered. PMID- 15342754 TI - Fractures of the radial head and neck treated with radial head excision. AB - BACKGROUND: The reported long-term outcomes of the treatment of radial head and neck fractures with excision of the radial head have been mixed. The purpose of the present study was to evaluate the long-term outcomes of primary or delayed radial head excision for the treatment of these fractures. METHODS: Sixty-one individuals (mean age, forty-four years) with thirty-nine Mason type-II, ten Mason type-III, and twelve Mason type-IV fractures were evaluated subjectively, objectively, and radiographically at a mean of eighteen years (range, eleven to thirty-three years) after treatment. Forty-three fractures were treated with primary radial head excision, and the remaining eighteen were treated with delayed radial head excision at a median of five months (range, one to 238 months) after the injury. RESULTS: At the time of follow-up, twenty-eight individuals had no symptoms, twenty-seven had occasional elbow pain, and six had daily pain. Four individuals with daily pain had had a Mason type-IV fracture. The range of motion of the formerly injured upper extremities was slightly less than that of the uninjured upper extremities in terms of flexion (139 degrees +/- 11 degrees compared with 142 degrees +/- 8 degrees ), extension (-7 degrees +/- 12 degrees compared with -1 degrees +/- 6 degrees ), and supination (77 degrees +/- 20 degrees compared with 85 degrees +/- 10 degrees ) (all p < 0.01). A higher percentage of formerly injured elbows than uninjured elbows had cysts, sclerosis, and osteophytes (73% compared with 7%; p < 0.001), but none had a reduced joint space. No differences were found between the outcomes for individuals treated with a primary radial head excision and those for individuals treated with a delayed excision. CONCLUSIONS: Following a displaced radial head or neck fracture, excision of the radial head often leads to a good or fair result. We found no differences in outcome between primary and delayed radial head excisions following a Mason type-II, III, or IV fracture. The outcomes are associated with the type of fracture, with Mason type-IV fractures having the worst results, rather than with the timing of the radial head excision (primary or delayed). PMID- 15342755 TI - Medial unicompartmental knee arthroplasty with the Miller-Galante prosthesis. AB - BACKGROUND: Unicompartmental knee arthroplasty has become a popular treatment alternative for osteoarthritis that is confined to the medial part of the knee. Excellent intermediate-term results recently have been reported in association with the Miller-Galante unicompartmental implant. The purpose of the present study was to report on our longer-term experience with the Miller-Galante medial unicompartmental knee implant. METHODS: We evaluated the results of 113 medial unicompartmental knee arthroplasties that had been performed with use of the Miller-Galante implant in eighty-four patients between 1989 and 2000. The mean age of the patients at the time of surgery was sixty-eight years. Forty-five patients were men, and thirty-nine were women. Thirteen patients (sixteen knees) died at a mean of seven years after the index arthroplasty. No patient was lost to follow-up. The remaining seventy-one patients (ninety-seven knees) were followed for a mean of ten years and were evaluated with use of the Knee Society clinical and radiographic rating system. RESULTS: Eleven knees were revised at a mean of four years after the index procedure. The mean Knee Society knee and function scores for the sixty-one patients (eighty-six knees) who were living and who had not had a revision improved from 48 and 53 points preoperatively to 93 and 80 points at the time of the most recent evaluation. The five and ten-year rates of survival were 94% and 90%, respectively, with revision to tricompartmental knee arthroplasty as the end point and 93% and 86%, respectively, with revision or radiographic loosening as the end point. CONCLUSIONS: The Miller-Galante medial unicompartmental knee arthroplasty provided excellent pain relief and restoration of function in carefully selected patients and demonstrated durable implant survival at ten years. PMID- 15342756 TI - Muscle and tendon morphology after reconstruction of the anterior cruciate ligament with autologous semitendinosus-gracilis graft. AB - BACKGROUND: The autologous semitendinosus-gracilis graft is the first choice of many orthopaedic surgeons when reconstructing the anterior cruciate ligament. The effect that graft harvest has on muscle and tendon morphology remains unclear. The purpose of this study was to describe these effects more completely. METHODS: Magnetic resonance images were acquired from eight patients before the anterior cruciate ligament reconstruction with semitendinosus-gracilis autograft and then again postoperatively after they had returned to sports. Muscle and tendon morphology was described by determining the volume and peak cross-sectional area of each structure on digitally reconstructed images. The effects that the procedure had on muscle and tendon length were evaluated separately and then together as a muscle-tendon complex. RESULTS: Anterior cruciate ligament reconstruction with semitendinosus-gracilis autograft resulted in a marked decrease in volume, cross-sectional area, and length of the semitendinosus and gracilis muscles. Tendon regeneration occurred in varying degrees in nearly all subjects. The morphology of the biceps femoris and semimembranosus muscles suggested that they had been compensating for the reduced semitendinosus and gracilis muscle function. Although semitendinosus and gracilis muscle retraction occurred following tendon stripping, nearly all of the subjects displayed evidence of at least partial tendon regeneration. CONCLUSIONS: Anterior cruciate ligament reconstruction with semitendinosus-gracilis autograft had a marked impact on semitendinosus and gracilis muscle morphology. However, this altered muscle morphology did not appear to have a clinically important impact on short term outcomes. The biceps femoris and semimembranosus muscles appear to compensate for reduced semitendinosus and gracilis function. Tendon regeneration is observed in most people, but it is often incomplete at six months. PMID- 15342757 TI - Magnetic resonance imaging after total hip arthroplasty: evaluation of periprosthetic soft tissue. AB - BACKGROUND: The evaluation of periprosthetic osteolysis in patients who have had a total hip arthroplasty is challenging, and traditional imaging techniques, including magnetic resonance imaging and computerized tomography, are limited by metallic artifact. The purpose of the present study was to investigate the use of modified magnetic resonance imaging techniques involving commercially available software to visualize periprosthetic soft tissues, to define the bone-implant interface, and to detect the location and extent of osteolysis. METHODS: Twenty eight hips in twenty-seven patients were examined to assess the extent of osteolysis (nineteen hips), enigmatic pain (five), heterotopic ossification (two), suspected tumor (one), or femoral nerve palsy (one). The results were correlated with conventional radiographic findings as well as with intraoperative findings (when available). RESULTS: Magnetic resonance imaging demonstrated the bone-implant interface and the surrounding soft-tissue envelope in all hips. Radiographs consistently underestimated the extent and location of acetabular osteolysis when compared with magnetic resonance imaging. Magnetic resonance imaging also disclosed radiographically occult extraosseous soft-tissue deposits that were similar in signal intensity to areas of osteolysis, demonstrated the relationship of these deposits to adjacent neurovascular structures, and allowed further visualization of hypertrophic synovial deposits that accompanied the bone resorption in twenty-five of the twenty-eight hips. CONCLUSIONS: Magnetic resonance imaging is effective for the assessment of the periprosthetic soft tissues in patients who have had a total hip arthroplasty. While not indicated for every patient who has pain at the site of an arthroplasty, these techniques can be effective for the evaluation of the surrounding soft-tissue envelope as well as intracapsular synovial deposits and are more effective than radiographs for the detection and evaluation of osteolysis, thus aiding in clinical management. PMID- 15342758 TI - Anterior release of the elbow for extension loss. AB - BACKGROUND: There are many causes of elbow contracture. When nonoperative techniques fail to increase the arc of motion of the elbow, surgical intervention may be indicated. The purpose of this study was to report the outcomes of surgical correction, predominantly with an anterior release, of elbow flexion contractures. In addition, we evaluated the efficacy of continuous passive motion in the immediate postoperative period. METHODS: We retrospectively reviewed the outcomes of 106 consecutive patients who had undergone anterior elbow release for the treatment of a flexion contracture between July 1975 and June 2001. Twenty nine patients were excluded because they had been followed for less than twelve months, leaving a study group of seventy-seven patients. Postoperatively, fifty four of the seventy-seven patients were treated with continuous passive motion and the other twenty-three patients were treated with extension splinting. The average duration of follow-up was thirty-three months. The average patient age was thirty-four years. The results were evaluated on the basis of both preoperative and postoperative radiographs as well as clinical measurements of elbow motion, all performed by the same examiner using the same large (47-cm long) goniometer. RESULTS: The mean preoperative extension in the seventy-seven patients was 52 degrees, which decreased to 20 degrees postoperatively. The mean flexion increased from 111 degrees preoperatively to 117 degrees postoperatively, and the mean total arc of motion increased from 59 degrees to 97 degrees. The total arc of motion in the patients treated with continuous passive motion increased 45 degrees, compared with an increase of 26 degrees in those treated with extension splinting. There were eleven complications in ten patients. The majority were traction neuropathies. There were two infections (one superficial and one deep), both of which resolved following treatment. CONCLUSIONS: Release of a pathologically thickened anterior elbow capsule through a predominantly anterior approach to correct diminished elbow extension is a safe and effective technique. Furthermore, compared with splinting in extension alone, the utilization of continuous passive motion during the postoperative period increases the total arc of motion. PMID- 15342759 TI - Recombinant human bone morphogenetic protein-2 delivered in an injectable calcium phosphate paste accelerates osteotomy-site healing in a nonhuman primate model. AB - BACKGROUND: In recent clinical trials demonstrating the efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2) for the acceleration of bone healing, investigators used carriers requiring open surgery for administration. In this study, we used a nonhuman primate fibular osteotomy model to evaluate injectable rhBMP-2/carrier formulations that can be administered in closed fractures. METHODS: The fibular osteotomy model was first characterized by evaluating surgically harvested fibular segments containing untreated osteotomy sites (controls) from seventy adult male Cynomolgus monkeys at eight weeks (twenty-four monkeys), ten weeks (thirty-four), twelve weeks (six), and fourteen weeks (six). Fibular segments, from twenty-four animals, in which an osteotomy had not been performed served as normal controls (intact). The contralateral limb of twenty-four of the animals was then used to evaluate the effect of rhBMP-2 administered, three hours after the osteotomy, in eight carrier formulations (buffer, calcium phosphate paste, and hyaluronan gel, hyaluronan paste, and gelatin foam formulated with and without tricalcium phosphate granules). Each carrier was used in three monkeys. At ten weeks, the fibulae with the treated osteotomy sites were harvested and were compared with the contralateral, untreated osteotomized fibulae (paired control). The most promising carrier, calcium phosphate paste (alpha bone substitute material, or alpha-BSM), was then evaluated in eleven additional animals. The outcomes included the findings on radiographs made weekly until the time of fibular harvest, the callus area, the biomechanical properties, and the histologic findings. RESULTS: Radiographic and histologic studies confirmed complete bridging of the control osteotomy sites in most animals by fourteen weeks. The mean torsional stiffness and maximum torque of the control osteotomy sites were 42.7% and 53.7%, 55.2% and 60.4%, 66.7% and 66.4% of the mean torsional stiffness and maximum torque of the intact fibulae at eight, ten, and twelve weeks, respectively, but they were not substantially different from the mean torsional stiffness and maximum torque of the intact fibulae at fourteen weeks (82.3% and 79.8%). In the carrier screening study, outcome measures of healing were more consistently enhanced in the rhBMP-2/alpha BSM-treated osteotomy sites. In the confirmatory study, the mean callus area, torsional stiffness, and maximum torque were 86%, 72%, and 68% greater in the rhBMP-2/alpha-BSM-treated osteotomy sites than in the paired-control osteotomy sites at ten weeks (p < 0.001). The torsional stiffness and maximum torque in the rhBMP-2/alpha-BSM-treated osteotomy sites were equal to those in the intact fibulae, whereas those parameters in the paired-control osteotomy sites were only 55% and 58%, respectively, of the torsional stiffness and maximum torque of the intact fibulae. Histologic analysis confirmed complete osseous bridging of the rhBMP-2/alpha-BSM-treated osteotomy sites but incomplete bridging of the paired control osteotomy sites at ten weeks. CONCLUSIONS: A single percutaneous injection of rhBMP-2/alpha-BSM accelerates the healing of fibular osteotomy sites in nonhuman primates by approximately 40% compared with the healing of untreated osteotomy sites. PMID- 15342760 TI - Effect of tendon release and delayed repair on the structure of the muscles of the rotator cuff: an experimental study in sheep. AB - BACKGROUND: Ruptures of the tendons of the rotator cuff lead to profound and possibly irreversible changes in the structure and physiological properties of the rotator cuff muscles. Muscle atrophy and fatty infiltration are important prognostic factors that affect the natural history and outcome of treatment. The purpose of this study was to examine the amount of muscle atrophy and fatty infiltration in an animal model and to determine whether the repair of a long standing tendon tear can reverse these changes. METHODS: The infraspinatus tendon in six sheep was released and encased in a silicone tube to prevent spontaneous healing. The musculotendinous unit was allowed to retract for forty weeks. Throughout this period, the muscular changes were studied with use of computed tomography, histological analysis, and electron microscopy. At forty weeks, the elasticity, intramuscular pressure, and perfusion were measured intraoperatively and a tendon repair was carried out. The structural changes of the muscle were studied for thirty-five weeks after the repair. The animals were then killed, and the musculotendinous units were examined macroscopically and by computed tomography, histological analysis, and electron microscopy. RESULTS: At the time of the tendon release, the infraspinatus showed no fatty changes. The force needed to cause a tendon excursion of 1 cm was a mean (and standard deviation) of 6.8 +/- 1 N. The application of tension on the tendon did not alter the perfusion and decreased the intramuscular pressure. After the tendon release, muscular atrophy developed and there was a significant increase (p < 0.001) in interfascicular and intrafascicular fat, representing fatty infiltration rather than fatty degeneration. Furthermore, there was an increase of interstitial connective tissue. At the time of the tendon repair, between forty and forty-two weeks after the release, there was a sevenfold poorer elasticity of the musculotendinous unit but preserved muscle perfusion. The structural changes increased six weeks after the repair and then recovered partially at twelve and thirty-five weeks thereafter but only to the amount demonstrated before the repair. CONCLUSIONS: Musculotendinous retraction induced by tendon release is associated with profound changes in the structure and function of the affected muscle. Vascularization, intramuscular pressure, and individual fiber composition are not markedly affected, and muscle fibers do not appear to degenerate. However, muscle atrophy, infiltration by fat cells, and an increase of interstitial connective tissue lead to impairment of the physiological properties of the muscle. These changes were irreversible under the conditions of this experiment with the repair technique used. PMID- 15342761 TI - Thirty-day mortality following hip arthroplasty for acute fracture. AB - BACKGROUND: Hip fractures are associated with a substantial mortality rate. Previous reports on perioperative mortality associated with hip arthroplasty for the treatment of acute fracture have not documented demographic and surgical characteristics that increase the likelihood of death. The purpose of the present study was to determine the prevalence of, and associated risk factors for, perioperative death following hip arthroplasty for the treatment of acute fracture. METHODS: Data were compiled from the computerized total joint registry at a single institution to determine the mortality rate following hip arthroplasty according to age, gender, diagnosis, implant type, and fixation mode. A review of this database revealed that 7774 consecutive patients had undergone hip arthroplasty for the treatment of an acute fracture between 1969 and 1997. The medical records of all patients who had died within thirty days after hip arthroplasty were reviewed retrospectively. RESULTS: The overall mortality rate within thirty days after hip arthroplasty for the treatment of an acute fracture was 2.4% (186 of 7774), yet notable variations in the mortality rate were seen within clinical subgroups. The thirty-day mortality rate was significantly higher for patients who had received a cemented implant, female patients, elderly patients, patients with cardiorespiratory comorbidities, and patients with intertrochanteric fractures. With the numbers available, there was no significant difference in mortality between patients who had been managed with total hip arthroplasty and those who had been managed with hemiarthroplasty. CONCLUSIONS: Hip arthroplasty for the diagnosis of acute fracture is associated with a nearly tenfold higher rate of perioperative mortality compared with elective hip arthroplasty. Medical optimization, appropriate choice of implants, and vigilant intraoperative management of these patients are essential. PMID- 15342762 TI - Two-stage revision hip arthroplasty for infection: comparison between the interim use of antibiotic-loaded cement beads and a spacer prosthesis. AB - BACKGROUND: A two-stage revision is a well-accepted method for the treatment of a deep infection of a hip with a joint implant. In the present study, the results associated with the interim use of antibiotic-loaded cement beads were compared with those associated with the interim use of an antibiotic-loaded cement prosthesis. METHODS: One hundred and twenty-eight consecutive patients who were managed with a two-stage revision hip arthroplasty for the treatment of an infection were followed clinically and radiographically for an average of 4.9 years. Cement beads were implanted following resection arthroplasty in the first seventy hips, and a custom cement prosthesis was implanted in the subsequent fifty-eight hips. RESULTS: There was no evidence of recurrent infection in 122 patients (95.3%); the infection-free rates in both groups were similar. The use of a spacer prosthesis was associated with a higher hip score, a shorter hospital stay, and better walking capacity in the interim period; a decreased operative time, less blood loss, and a lower transfusion requirement at the time of reimplantation; and fewer postoperative dislocations. CONCLUSIONS: The present study supports the safety and efficacy of the routine use of an antibiotic-loaded cement prosthesis in the interim between the stages of a two-stage revision procedure for the treatment of an infection at the site of a hip arthroplasty. PMID- 15342763 TI - Clinical and radiographic results associated with a modern, cementless modular cup design in total hip arthroplasty. AB - BACKGROUND: First-generation cementless modular cups reproducibly achieved fixation to bone but were associated with unacceptable rates of pelvic osteolysis and mechanical failure. Consequently, second-generation cups were developed with shells that had a limited number of holes (or no holes) as well as improved locking mechanisms, a polished inner surface, and increased conformity with the liner. The purpose of the present study was to evaluate the clinical and radiographic results associated with the use of a second-generation acetabular component for primary total hip arthroplasty. METHODS: Two hundred and ninety seven patients underwent 335 consecutive primary total hip arthroplasties that were performed by a single surgeon with a second-generation modular acetabular component. All cups were implanted with a press-fit technique. Ten patients were lost to follow-up, and sixteen died from unrelated causes. The remaining 271 patients (308 hips) were followed clinically (with the Hospital for Special Surgery hip-scoring system) and radiographically for four to seven years. RESULTS: One cup was revised because of aseptic loosening. There were seven additional revisions: five were performed because of aseptic loosening of the stem with a well-fixed cup, and two were performed because of deep infection. Among the 271 patients who were alive at the time of the last follow-up, 266 (98%) had retention of the cup and 264 (97%) had retention of both components with a good or excellent clinical result. In the group of 229 patients (262 hips) with complete radiographic follow-up, 259 cups were well fixed and the average wear rate (for the 246 hips for which this rate could be calculated) was 0.09 mm/yr. Osteolysis was detected in twelve hips (5%) and was associated with male gender (p = 0.001) and the annual wear rate (p = 0.004). The extent of calcar resorption was also associated with the annual wear rate (p < 0.001). CONCLUSIONS: This second-generation acetabular cup design predictably achieved bone fixation and was associated with low rates of revision for loosening and osteolysis after intermediate-term follow-up. PMID- 15342764 TI - Operative correction of adolescent idiopathic scoliosis in male patients. A radiographic and functional outcome comparison with female patients. AB - BACKGROUND: The outcomes following surgical treatment of adolescent idiopathic scoliosis have traditionally been assessed on the basis of radiographic parameters and, more recently, functional outcome measures. However, we know of no published studies in which radiographic and functional outcomes following surgery were compared between male and female patients. METHODS: Fifty-two male patients who had had surgery for adolescent idiopathic scoliosis were compared retrospectively with two groups of female patients: a random sample of 130 female patients who had had surgical treatment for adolescent idiopathic scoliosis during the same time period and a subgroup of fifty-two of these female patients who had been matched to the male patients with regard to curve type and magnitude. Radiographic parameters were compared between the male and female patients, and the Scoliosis Research Society outcome questionnaire was used to compare functional results between the male patients and the matched female group. RESULTS: Compared with the random sample of female patients, the male patients were older at the time of presentation (average [and standard deviation], 13.9 +/- 1.9 compared with 12.8 +/- 1.4 years) and at the time of surgery (average, 15.9 +/- 2.0 compared with 14.2 +/- 1.4 years) (p < 0.05). The male and female patients presented with primary coronal curves of similar magnitudes (average, 48 degrees +/- 19 degrees compared with 47 degrees +/- 13 degrees ), but the male patients had larger curves at the time of surgery (average, 62 degrees +/- 11 degrees compared with 56 degrees +/- 10 degrees ) with greater coronal plane imbalance. Compared with the subset of female patients matched for curve type and magnitude, the male patients had, on average, a longer surgical time (263 +/- 61 compared with 202 +/- 40 minutes), greater intraoperative blood loss (1148 +/- 660 compared with 944 +/- 408 mL), and less curve correction in the coronal plane (from 62 degrees +/- 11 degrees to 31 degrees +/- 11 degrees compared with from 59 degrees +/- 10 degrees to 23 degrees +/- 9 degrees ) (p < 0.05). The loss of coronal plane correction, the final coronal balance, all measured sagittal plane parameters, and the prevalence of complications were the same in these two groups. With regard to functional outcome, the scores were similar with the exception that the male patients had lower scores in the category of "function from back condition" when compared with the matched female patients (3.8 +/- 0.2 compared with 4.3 +/- 0.3 points) (p < 0.05). CONCLUSIONS: Adolescent idiopathic scoliosis is identified at a later age in male patients than in female patients with similar curve types. The curve magnitudes in the male patients are greater at the time of surgery. When surgeons are planning operative correction of adolescent idiopathic scoliosis in male patients, they should expect longer operative time, greater blood loss, and less coronal plane correction of the primary curve. However, balance in the coronal and sagittal planes should be achieved and complication rates and functional outcomes can be expected to be similar to those in female patients. PMID- 15342765 TI - Orientation of the femoral component in surface arthroplasty of the hip. A biomechanical and clinical analysis. AB - BACKGROUND: Although the orientation of the femoral component has been shown to influence the outcome of total hip replacement, its effect on the clinical outcome of surface arthroplasty has not been studied, to our knowledge. The purpose of this study was to examine the relationship between femoral component positioning and the outcome of a surface arthroplasty of the hip. METHODS: We reviewed the results of ninety-four hybrid metal-on-metal surface arthroplasties in patients who were forty years old or younger at the time of the operation and were followed for a minimum of two years or until the prosthesis failed. Measurements of the hip reconstruction were made on the anteroposterior pelvic radiograph. The correlation between the orientation of the femoral component and the outcome of the arthroplasty was evaluated, as were stresses within the resurfaced femoral head as a function of the orientation of the femoral component. RESULTS: The mean duration of follow-up was 4.2 years. Thirteen hips had an adverse outcome, defined as conversion to a total hip replacement, radiolucency of >1 mm in thickness adjacent to the femoral stem, or narrowing of the femoral neck of >10%. The mean femoral stem-shaft angle in the coronal plane was 138 degrees, with the hips that had an adverse outcome having a significantly lower mean angle than the rest of the cohort (133 degrees compared with 139 degrees, p = 0.03). Hips with an angle of 10 kb) of AhR responsive genes were analyzed by the genetic algorithm and a variety of other computational methods. Rules were established on the local oligonucleotide context in the flanks of the AhR binding sites, on the occurrence of clusters of AhR recognition elements, and on the presence in the promoters of specific combinations of multiple binding sites for the transcription factors cooperating in the AhR regulatory network. Our rules were applied to search for yet unknown Ah-receptor target genes. Experimental evidence is presented to demonstrate high fidelity of this novel in silico approach. PMID- 15342793 TI - Protocols for regulation and study of diphosphoinositol polyphosphates. AB - The roles of diphosphoinositol polyphosphates (DIPs) in mammalian cell biology have been difficult to determine because of the lack of tools known to regulate their levels. I have determined a series of protocols that regulate these DIPs, and these can be used to further our understanding of these molecules. Sorbitol and sucrose significantly raised levels of bis-diphosphoinositol tetrakisphosphate ([PP]2-InsP4) but slightly lowered levels of diphosphoinositol pentakisphosphate (PP-InsP5) in DDT1 MF-2 cells. These effects correlate with the ability of hyperosmotic stress to interfere with protein trafficking described previously and suggest that [PP]2-InsP4 specifically impedes protein trafficking. The effects on [PP]2-InsP4 were not regulated by extracellular signal-regulated kinase or phospholipase D, as exemplified by the lack of effect of U0126 and butan-1-ol. I have also found that genistein potently and rapidly lowers levels of [PP]2-InsP4, whereas a similar inhibitor, herbimycin, was without effect. Thapsigargin, a sarcoplasmic-endoplasmic reticulum Ca(2+)-ATPase pump inhibitor previously shown to selectively lower PP-InsP5 after short-term treatment, also selectively raises PP-InsP5 after a longer treatment. The calmodulin inhibitors N (6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) and chlorpromazine significantly lowered all higher inositol phosphates, as well as DIPs, whereas the calmodulin-dependent kinase inhibitors methyl 9-(S)-12-(R)-epoxy-1H diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-i][1,6]benzodiazocine-2,3,9,10,11,12 hexahydro-10-(R)hydroxy-9-methyl-1-oxo-10-carboxylate (K-252a) and 2-[N-(2 hydroxyethyl)-N-(4-methoxybenzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N methylbenzylamine (KN-93) were without effect. W-7 and chlorpromazine also lowered levels of phosphatidylinositol 4,5-bisphosphate and ATP but greatly increased levels of phosphatidylinositol 4-phosphate. Trypan blue exclusion deemed that these doses were not cytotoxic. These results identify an increasing number of reagents that regulate DIP levels. Using these tools, and those described previously, we can further understand the roles of the DIPs in cell biology. PMID- 15342794 TI - Analysis of ATP-binding cassette transporter expression in drug-selected cell lines by a microarray dedicated to multidrug resistance. AB - Discovery of the multidrug resistance protein 1 (MDR1), an ATP-binding cassette (ABC) transporter able to transport many anticancer drugs, was a clinically relevant breakthrough in multidrug resistance research. Although the overexpression of ABC transporters such as P-glycoprotein/ABCB1, MRP1/ABCC1, and MXR/ABCG2 seems to be a major cause of failure in the treatment of cancer, acquired resistance to multiple anticancer drugs may also be multifactorial, involving alteration of detoxification processes, apoptosis, DNA repair, drug uptake, and overexpression of other ABC transporters. As a tool for the study of such phenomena, we designed and created a microarray platform, the ABC-ToxChip, to evaluate relative levels of transcriptional activation among genes involved in the various mechanisms of resistance. In the ABC-ToxChip, a comprehensive set of genes important in toxicological responses (represented by 2200 cDNA probes) is complemented with probes specifically matching ABC transporters as well as oligonucleotides representing 18,000 unique human genes. By comparing the transcriptional profiles of KB-3-1 and DU-145 parental cells with resistant derivatives selected in colchicine (KB-8-5), and 9-nitro-camptothecin (RCO.1), respectively, we demonstrate that ABC transporters (ABCB1/MDR1 and ABCC2/MRP2, respectively) show dramatic overexpression, whereas the glutathione S-transferase gene GST-Pi shows the strongest decrease in expression among the 20,000 genes studied. The results were confirmed by quantitative reverse transcription polymerase chain reaction and immunohistochemistry. The custom-designed ABC-Tox microarray presented here will be helpful to elucidate mechanisms leading to anticancer drug resistance. PMID- 15342795 TI - Different regulatory mechanisms underlie similar transposable element profiles in pufferfish and fruitflies. AB - Comparative analysis of recently sequenced eukaryotic genomes has uncovered extensive variation in transposable element (TE) abundance, diversity, and distribution. The TE profile in the sequenced pufferfish genomes is more similar to that of Drosophila melanogaster than to human or mouse, in that pufferfish TEs exhibit low overall abundance, high family diversity, and localization in the heterochromatin. It has been suggested that selection against the deleterious effects of ectopic recombination between TEs has structured the TE profile in Drosophila and pufferfish but not in humans. We test this hypothesis by measuring the sample frequency of 48 euchromatic TE insertions in the genome of the green spotted pufferfish (Tetraodon nigroviridis). We estimate the strength of selection acting on recent insertions by analyzing the site frequency spectrum using a maximum-likelihood approach. We show that in contrast to Drosophila, euchromatic TE insertions in Tetraodon are selectively neutral and that the low copy number and compartmentalized distribution of TEs in the Tetraodon genome must be caused by regulation by means other than purifying selection acting on recent insertions. Inference of regulatory processes governing TE profiles should take into account factors such as effective population size, incidence of inbreeding/outcrossing, and other species-specific traits. PMID- 15342796 TI - A broad survey of recombination in animal mitochondria. AB - Recombination in mitochondrial DNA (mtDNA) remains a controversial topic. Here we present a survey of 279 animal mtDNA data sets, of which 12 were from asexual species. Using four separate tests, we show that there is widespread evidence of recombination; for one test as many as 14.2% of the data sets reject a model of clonal inheritance and in several data sets, including primates, the recombinants can be identified visually. We show that none of the tests give significant results for obligate clonal species (apomictic pathogens) and that the sexual species show significantly greater evidence of recombination than asexual species. For some data sets, such as Macaca nemestrina, additional data sets suggest that the recombinants are not artifacts. For others, it cannot be determined whether the recombinants are real or produced by laboratory error. Either way, the results have important implications for how mtDNA is sequenced and used. PMID- 15342797 TI - Evolution of vertebrate genes related to prion and Shadoo proteins--clues from comparative genomic analysis. AB - Recent findings of new genes in fish related to the prion protein (PrP) gene PRNP, including our recent report of SPRN coding for Shadoo (Sho) protein found also in mammals, raise issues of their function and evolution. Here we report additional novel fish genes found in public databases, including a duplicated SPRN gene, SPRNB, in Fugu, Tetraodon, carp, and zebrafish encoding the Sho2 protein, and we use comparative genomic analysis to analyze the evolutionary relationships and to infer evolutionary trajectories of the complete data set. Phylogenetic footprinting performed on aligned human, mouse, and Fugu SPRN genes to define candidate regulatory promoter regions, detected 16 conserved motifs, three of which are known transcription factor-binding sites for a receptor and transcription factors specific to or associated with expression in brain. This result and other homology-based (VISTA global genomic alignment; protein sequence alignment and phylogenetics) and context-dependent (genomic context; relative gene order and orientation) criteria indicate fish and mammalian SPRN genes are orthologous and suggest a strongly conserved basic function in brain. Whereas tetrapod PRNPs share context with the analogous stPrP-2-coding gene in fish, their sequences are diverged, suggesting that the tetrapod and fish genes are likely to have significantly different functions. Phylogenetic analysis predicts the SPRN/SPRNB duplication occurred before divergence of fish from tetrapods, whereas that of stPrP-1 and stPrP-2 occurred in fish. Whereas Sho appears to have a conserved function in vertebrate brain, PrP seems to have an adaptive role fine tuned in a lineage-specific fashion. An evolutionary model consistent with our findings and literature knowledge is proposed that has an ancestral prevertebrate SPRN-like gene leading to all vertebrate PrP-related and Sho-related genes. This provides a new framework for exploring the evolution of this unusual family of proteins and for searching for members in other fish branches and intermediate vertebrate groups. PMID- 15342798 TI - Adaptive evolution drives the diversification of zinc-finger binding domains. AB - The human genome is estimated to contain 700 zinc-finger genes, which perform many key functions, including regulating transcription. The dramatic increase in the number of these genes as we move from yeast to C. elegans to Drosophila and to humans, as well as the clustered organization of these genes in humans, suggests that gene duplication has played an important role in expanding this family of genes. Using likelihood methods developed by Yang and parsimony methods introduced by Suzuki and Gojobori, we have investigated four clusters of zinc finger genes on human chromosome 19 and found evidence that positive selection was involved in diversifying the family of zinc-finger binding motifs. PMID- 15342799 TI - Culture and depression. PMID- 15342800 TI - Cultural competence--marginal or mainstream movement? PMID- 15342801 TI - Conflicts of interest at the NIH--resolving the problem. PMID- 15342802 TI - SIDS--a syndrome in search of a cause. PMID- 15342803 TI - Body fat, leptin, and hypothalamic amenorrhea. PMID- 15342804 TI - Tamoxifen with or without breast irradiation in women 50 years of age or older with early breast cancer. AB - BACKGROUND: We determined the effect of breast irradiation plus tamoxifen on disease-free survival and local relapse in women 50 years of age or older who had T1 or T2 node-negative breast cancer. METHODS: Between December 1992 and June 2000, 769 women with early breast cancer (tumor diameter, 5 cm or less) were randomly assigned to receive breast irradiation plus tamoxifen (386 women) or tamoxifen alone (383 women). The median follow-up was 5.6 years. RESULTS: The rate of local relapse at five years was 7.7 percent in the tamoxifen group and 0.6 percent in the group given tamoxifen plus irradiation (hazard ratio, 8.3; 95 percent confidence interval, 3.3 to 21.2; P<0.001), with corresponding five-year disease-free survival rates of 84 percent and 91 percent (P=0.004). A planned subgroup analysis of 611 women with T1, receptor-positive tumors indicated a benefit from radiotherapy (five-year rates of local relapse, 0.4 percent with tamoxifen plus radiotherapy and 5.9 percent with tamoxifen alone; P<0.001). Overall, there was a significant difference in the rate of axillary relapse at five years (2.5 percent in the tamoxifen group and 0.5 percent in the group given tamoxifen plus irradiation, P=0.049), but no significant difference in the rates of distant relapse or overall survival. CONCLUSIONS: As compared with tamoxifen alone, radiotherapy plus tamoxifen significantly reduces the risk of breast and axillary recurrence after lumpectomy in women with small, node-negative, hormone receptor-positive breast cancers. PMID- 15342805 TI - Lumpectomy plus tamoxifen with or without irradiation in women 70 years of age or older with early breast cancer. AB - BACKGROUND: In women 70 years of age or older who have early breast cancer, it is unclear whether lumpectomy plus tamoxifen is as effective as lumpectomy followed by tamoxifen plus radiation therapy. METHODS: Between July 1994 and February 1999, we randomly assigned 636 women who were 70 years of age or older and who had clinical stage I (T1N0M0 according to the tumor-node-metastasis classification), estrogen-receptor-positive breast carcinoma treated by lumpectomy to receive tamoxifen plus radiation therapy (317 women) or tamoxifen alone (319 women). Primary end points were the time to local or regional recurrence, the frequency of mastectomy for recurrence, breast-cancer-specific survival, the time to distant metastasis, and overall survival. RESULTS: The only significant difference between the two groups was in the rate of local or regional recurrence at five years (1 percent in the group given tamoxifen plus irradiation and 4 percent in the group given tamoxifen alone, P<0.001). There were no significant differences between the two groups with regard to the rates of mastectomy for local recurrence, distant metastases, or five-year rates of overall survival (87 percent in the group given tamoxifen plus irradiation and 86 percent in the tamoxifen group, P=0.94). Assessment by physicians and patients of cosmetic results and adverse events uniformly rated tamoxifen plus irradiation inferior to tamoxifen alone. CONCLUSIONS: Lumpectomy plus adjuvant therapy with tamoxifen alone is a realistic choice for the treatment of women 70 years of age or older who have early, estrogen-receptor-positive breast cancer. PMID- 15342806 TI - Second-trimester maternal serum levels of alpha-fetoprotein and the subsequent risk of sudden infant death syndrome. AB - BACKGROUND: Unexplained stillbirth and the sudden infant death syndrome (SIDS) share some features. A raised maternal serum level of alpha-fetoprotein during the second trimester of pregnancy is a marker of placental dysfunction and a strong predictor of the risk of unexplained stillbirth. It is unknown whether alpha-fetoprotein levels also predict the risk of SIDS. METHODS: We linked a prenatal-screening database for women in western Scotland with databases of maternity, perinatal death, and birth and death certifications to assess the association between second-trimester levels of maternal serum alpha-fetoprotein and the subsequent risk of SIDS. RESULTS: Among 214,532 women with singleton births, there were 114 cases of SIDS (incidence, 2.7 per 10,000 births among women with alpha-fetoprotein levels in the lowest quintile and 7.5 per 10,000 births among those with levels in the highest quintile). When the lowest quintile was used as a referent, the unadjusted odds ratios for SIDS for the second through fifth quintiles were 1.7 (95 percent confidence interval, 0.8 to 3.5), 1.8 (95 percent confidence interval, 0.9 to 3.7), 2.5 (95 percent confidence interval, 1.3 to 4.8), and 2.8 (95 percent confidence interval, 1.4 to 5.4), respectively (P for trend = 0.001). The risk of SIDS varied inversely with the birth-weight percentile and the gestational age at delivery; after adjustment for these factors, the odds ratios for SIDS were 1.7 (95 percent confidence interval, 0.8 to 3.5), 1.7 (95 percent confidence interval, 0.8 to 3.5), 2.2 (95 percent confidence interval, 1.1 to 4.4), and 2.2 (95 percent confidence interval, 1.1 to 4.3), respectively (P for trend = 0.01). CONCLUSIONS: There is a direct association between second-trimester maternal serum alpha-fetoprotein levels and the risk of SIDS, which may be mediated in part through impaired fetal growth and preterm birth. PMID- 15342807 TI - Recombinant human leptin in women with hypothalamic amenorrhea. AB - BACKGROUND: Disruptions in hypothalamic-gonadal and other endocrine axes due to energy deficits are associated with low levels of the adipocyte-secreted hormone leptin and may result in hypothalamic amenorrhea. We hypothesized that exogenous recombinant leptin replacement would improve reproductive and neuroendocrine function in women with hypothalamic amenorrhea. METHODS: Eight women with hypothalamic amenorrhea due to strenuous exercise or low weight were studied for one month before receiving recombinant human leptin and then while receiving treatment for up to three months. Six control subjects with hypothalamic amenorrhea received no treatment and were studied for a mean (+/-SD) of 8.5+/-8.1 months. RESULTS: Luteinizing hormone (LH) pulsatility, body weight, ovarian variables, and hormone levels did not change significantly over time in the controls and during a one-month control period before recombinant leptin therapy in the treated subjects. In contrast, recombinant leptin treatment increased mean LH levels and LH pulse frequency after two weeks and increased maximal follicular diameter, the number of dominant follicles, ovarian volume, and estradiol levels over a period of three months. Three patients had an ovulatory menstrual cycle (P<0.05 for the comparison with an expected rate of spontaneous ovulation of 10 percent); two others had preovulatory follicular development and withdrawal bleeding during treatment (P<0.05). Recombinant leptin significantly increased levels of free triiodothyronine, free thyroxine, insulin-like growth factor 1, insulin-like growth factor-binding protein 3, bone alkaline phosphatase, and osteocalcin but not cortisol, corticotropin, or urinary N-telopeptide. CONCLUSIONS: Leptin administration for the relative leptin deficiency in women with hypothalamic amenorrhea appears to improve reproductive, thyroid, and growth hormone axes and markers of bone formation, suggesting that leptin, a peripheral signal reflecting the adequacy of energy stores, is required for normal reproductive and neuroendocrine function. PMID- 15342808 TI - Management of cutaneous melanoma. PMID- 15342809 TI - Images in clinical medicine. Detection of liver abscesses by T1-weighted magnetic resonance imaging. PMID- 15342810 TI - Clinical problem-solving. Undercover and overlooked. PMID- 15342811 TI - Breast radiotherapy after lumpectomy--no longer always necessary. PMID- 15342812 TI - No pancreatic endocrine stem cells? PMID- 15342813 TI - Homocysteine as a predictive factor for hip fracture in older persons. PMID- 15342814 TI - Myostatin mutation associated with gross muscle hypertrophy in a child. PMID- 15342815 TI - Plan B--the FDA and emergency contraception. PMID- 15342816 TI - Hypoparathyroidism and autoimmune polyendocrine syndromes. PMID- 15342817 TI - Case 18-2004: a 61-year-old man with rectal bleeding. PMID- 15342818 TI - Imatinib mesylate for cerebral Langerhans'-cell histiocytosis. PMID- 15342819 TI - Images in clinical medicine. Bilateral renal arteriovenous malformation. PMID- 15342820 TI - Asthma update: epidemiology and pathophysiology. PMID- 15342821 TI - Domestic violence: a pediatric concern. PMID- 15342822 TI - Periorbital and orbital infections. PMID- 15342823 TI - Index of suspicion. PMID- 15342824 TI - Thimerosal exposure in infants and developmental disorders: a prospective cohort study in the United kingdom does not support a causal association. AB - OBJECTIVE: There is an established link between exposure to mercury and impaired childhood cognitive development and early motor skills. Thimerosal (also known as thiomersal), a preservative used in a number of children's vaccines, contains ethylmercury (an organic compound of mercury), and there has been concern that this exposure to mercury may be of some detriment to young children. The aim of this research was to test in a large United Kingdom population-based cohort whether there is any evidence to justify such concerns. METHODS: We used population data from a longitudinal study on childhood health and development. The study has been monitoring >14,000 children who are from the geographic area formerly known as Avon, United Kingdom, and were delivered in 1991-1992. The age at which doses of thimerosal-containing vaccines were administered was recorded, and measures of mercury exposure by 3, 4, and 6 months of age were calculated and compared with a number of measures of childhood cognitive and behavioral development covering the period from 6 to 91 months of age. RESULTS: Contrary to expectation, it was common for the unadjusted results to suggest a beneficial effect of thimerosal exposure. For example, exposure at 3 months was inversely associated with hyperactivity and conduct problems at 47 months; motor development at 6 months and at 30 months; difficulties with sounds at 81 months; and speech therapy, special needs, and "statementing" at 91 months. After adjustment for birth weight, gestation, gender, maternal education, parity, housing tenure, maternal smoking, breastfeeding, and ethnic origins, we found 1 result of 69 to be in the direction hypothesized-poor prosocial behavior at 47 months was associated with exposure by 3 months of age (odds ratio: 1.12; 95% confidence interval: 1.01-1.23) compared with 8 results that still supported a beneficial effect. CONCLUSIONS: We could find no convincing evidence that early exposure to thimerosal had any deleterious effect on neurologic or psychological outcome. PMID- 15342825 TI - Thimerosal exposure in infants and developmental disorders: a retrospective cohort study in the United kingdom does not support a causal association. AB - OBJECTIVE: After concerns about the possible toxicity of thimerosal-containing vaccines in the United States, this study was designed to investigate whether there is a relationship between the amount of thimerosal that an infant receives via diphtheria-tetanus-whole-cell pertussis (DTP) or diphtheria-tetanus (DT) vaccination at a young age and subsequent neurodevelopmental disorders. METHODS: A retrospective cohort study was performed using 109 863 children who were born from 1988 to 1997 and were registered in general practices in the United Kingdom that contributed to a research database. The disorders investigated were general developmental disorders, language or speech delay, tics, attention-deficit disorder, autism, unspecified developmental delays, behavior problems, encopresis, and enuresis. Exposure was defined according to the number of DTP/DT doses received by 3 and 4 months of age and also the cumulative age-specific DTP/DT exposure by 6 months. Each DTP/DT dose of vaccine contains 50 microg of thimerosal (25 microg of ethyl mercury). Hazard ratios (HRs) for the disorders were calculated per dose of DTP/DT vaccine or per unit of cumulative DTP/DT exposure. RESULTS: Only in 1 analysis for tics was there some evidence of a higher risk with increasing doses (Cox's HR: 1.50 per dose at 4 months; 95% confidence interval [CI]: 1.02-2.20). Statistically significant negative associations with increasing doses at 4 months were found for general developmental disorders (HR: 0.87; 95% CI: 0.81-0.93), unspecified developmental delay (HR: 0.80; 95% CI: 0.69-0.92), and attention-deficit disorder (HR: 0.79; 95% CI: 0.64-0.98). For the other disorders, there was no evidence of an association with thimerosal exposure. CONCLUSIONS: With the possible exception of tics, there was no evidence that thimerosal exposure via DTP/DT vaccines causes neurodevelopmental disorders. PMID- 15342826 TI - Sleep of excessively crying infants: a 24-Hour Ambulatory Sleep Polygraphy study. AB - OBJECTIVE: Parents' reports suggest that excessively crying or colicky infants sleep less compared with control subjects. The aim of this study was to determine the sleep-wake structure of excessively crying infants throughout a 24-hour cycle. METHODS: A 24-hour sleep polygraphy study was conducted at home for 24 excessively crying infants and 23 control subjects at the age of 6 weeks. In addition, parental diaries were kept for 4 days. RESULTS: In sleep polygraphy recordings, no major differences between study groups were observed in either the duration or the structure of the 24-hour sleep. In the diaries, the parents overestimated the amount of sleep in both study groups. The parents of the control infants overestimated the amount of sleep more than the parents of excessively crying infants (69.8 minutes [standard deviation: 79.3] compared with 27.1 minutes [standard deviation: 65.4], respectively). In excessively crying infants, the proportion of rapid eye movement sleep was higher during the 3-hour period from the beginning of the first long sleep in the evening and lower during the preceding 3-hour period compared with the control group. CONCLUSIONS: The results of this study suggest that diary-based studies are prone to be biased as the parents of the control infants are more likely to overestimate the amount of infant's sleep and, therefore, report more sleep than the parents of the crying infants. Although no differences in the total amount of sleep or proportions of sleep stages were observed, excessively crying infants may be characterized by a disturbance that affects rapid eye movement and non-rapid eye movement sleep stage proportion during evening hours. PMID- 15342827 TI - Diagnosis and treatment of behavioral health disorders in pediatric practice. AB - OBJECTIVE: There has been a strong push toward the recognition and treatment of children with behavioral health problems by primary care pediatricians. This study was designed to assess the extent to which a sample of primary care pediatricians diagnose and treat behavioral health problems and to identify factors that may contribute to their behavioral health practice. METHODS: A standard interview was conducted with 47 pediatricians who work in primary care settings in a predominantly urban setting in North Carolina. Pediatricians' responses to questions about the estimated percentage of children in their practice with a behavioral health disorder, tools used to make diagnoses, frequent and infrequent diagnoses made, comfort level with making a diagnosis, reasons for not making a diagnosis, use of psychotropic medications, types of nonmedication interventions provided, educational background, and needs involving behavioral health issues were evaluated. RESULTS: Pediatricians estimated that the average percentage of children in their practices with a behavioral health disorder was 15%. The study did not find significant differences in perceptions related to time in practice or gender of the pediatric provider. The most frequent behavioral health diagnosis was attention-deficit/hyperactivity disorder (ADHD), and the majority incorporated behavioral questionnaires, expressed a high level of comfort with the diagnosis, and frequently or occasionally prescribed stimulants. Variability was noted in both practice and comfort for other behavioral health disorders. Slightly fewer than half of the pediatricians frequently diagnosed anxiety and depression. Those who make these diagnoses commonly incorporated questionnaires and reported frequent or occasional use of selective serotonin reuptake inhibitors. Comfort in making the diagnosis of anxiety was highly associated with use of selective serotonin reuptake inhibitors. The vast majority (96%) of pediatricians provided nonmedication interventions, including supportive counseling, education for coping with ADHD, behavior modification, and/or stress management. Diagnosis and treatment of severe behavioral health disorders were infrequent throughout the pediatric practices. Areas of greatest educational interest included psychopharmacology, diagnosis and treatment of depression and anxiety, and updates on ADHD. The majority of pediatric providers did not identify a need for education about several high-prevalence disorders that they do not frequently diagnose or treat, including conduct disorder and substance abuse. CONCLUSIONS: Pediatricians in this sample frequently diagnosed and treated ADHD. For all other behavioral health disorders, pediatricians reported variability in both comfort and practice. They frequently provided both pharmacologic and nonpharmacologic treatments for children and adolescents with mild to moderate behavioral health disorders but not for severe disorders. Although they identified needs for additional education for anxiety and depression, the majority did not identify educational needs for several high-prevalence behavioral health disorders, including conduct disorder and substance abuse. PMID- 15342828 TI - Effects of atopic dermatitis on young American children and their families. AB - OBJECTIVE: The psychologic, physical, and social impact of atopic dermatitis is complex and varies among children of different ages, and the effects of atopic dermatitis on the quality of life of very young American children and their families are not well understood. This study was conducted to document these effects of atopic dermatitis on young children and their families. METHODS: Directed focus sessions were performed with parents of 26 young children with atopic dermatitis and 6 expert clinicians. Specific mentions of the ways in which atopic dermatitis affected the children and parents were reviewed, rank ordered, and categorized according to similarity in content. The categories were examined to determine the domains represented, and the domains were used to compose a conceptual framework of all of the ways that atopic dermatitis affects children and their families. RESULTS: Parents and experts mentioned a total of 181 specific quality-of-life effects. A conceptual framework, developed from the 181 effects, contains the domains of physical health, emotional health, physical functioning, and social functioning; each domain includes effects on both the child and the parents. CONCLUSIONS: Atopic dermatitis greatly affects the quality of life of afflicted children and their families. The comprehensive conceptual framework summarizes the ways in which atopic dermatitis affects the quality of life in young American children. This conceptual framework forms the basis from which quality-of-life instruments can be developed. PMID- 15342829 TI - Perinatal stroke in children with motor impairment: a population-based study. AB - OBJECTIVE: Risk factors for perinatal arterial stroke (PAS) are poorly understood. Most previous studies lack an appropriate control group and include only infants with symptoms in the newborn period. We set out to determine prenatal and perinatal risk factors for PAS. METHODS: In a population-based, case control study nested within the cohort of 231 582 singleton infants who were born at >or=36 weeks' gestation in Northern California Kaiser hospitals from 1991 to 1998, we searched electronically for children with motor impairment and reviewed their medical records to identify diagnoses of PAS. Control subjects were randomly selected from the study population. A medical record abstractor reviewed delivery records without knowledge of case status. RESULTS: The prevalence of PAS with motor impairment was 17/100,000 live births. Of 38 cases, 26 (68%) presented after 3 months of age with hemiparesis or seizures. All 12 newborns with acute stroke symptoms had seizures. A delayed presentation was more common in children with moderate to severe motor impairment than among infants with only mild motor abnormalities (24 of 31 vs 2 of 7). Prepartum risk factors significantly associated with PAS in multivariate analysis were preeclampsia (odds ratio [OR]: 3.6; 95% confidence interval [CI]: 1.1-11.4) and intrauterine growth restriction (OR: 5.3; 95% CI: 1.5-18.6). Newborns with PAS were also at higher risk of delivery complications, such as emergency cesarean section (OR: 6.8; 95% CI: 2.7 16.6), 5-minute Apgar <7 (OR: 23.6; 95% CI: 4.1-237), and resuscitation at birth (OR: 4.5; 95% CI: 1.6-12.3). CONCLUSIONS: Preeclampsia and intrauterine growth restriction (IUGR) may be independent risk factors for perinatal stroke resulting in motor impairment. Large multicenter studies that include all children with perinatal stroke are needed to determine further the risk factors and outcome of perinatal stroke. PMID- 15342830 TI - Effect of botulinum toxin in the treatment of drooling: a controlled clinical trial. AB - OBJECTIVE: To investigate the clinical effectiveness of botulinum neurotoxin type A (BoNT) to reduce drooling in children with cerebral palsy (CP). METHODS: A controlled clinical trial was performed in which the results of single-dose BoNT injections in the submandibular glands were compared with treatment with scopolamine. Forty-five children who had CP and experienced severe drooling were enrolled. Drooling severity was measured at baseline, during application of scopolamine, and at different intervals after BoNT injections up to 24 weeks, using the Drooling Quotient (DQ), the Teacher Drooling Scale (TDS), and Visual Analog Scales (VAS). RESULTS: Drooling was reduced during scopolamine application as well as after BoNT injections. Compared with baseline, the mean DQ showed a significant decrease throughout the study. Greatest reductions were achieved 2 to 8 weeks after BoNT injection. No significant differences were found between scopolamine measurements and those up to 24 months after BoNT injection. Using VAS, parents recorded the effect on drooling in which significant differences were found between baseline VAS score and all follow-up assessments. According to our definition of "success to therapy," demanding a 2-point decrease on the TDS, 61.5% of patients responded to BoNT injections. Analysis of the DQ demonstrated a response rate of 53% of the patients to scopolamine and 48.7% to BoNT until 24 weeks after BoNT injections, the actual duration of this study. As a reaction to scopolamine, 71.1% of the patients had moderate to severe side effects. Only nonsevere, incidental side effects were reported after BoNT injections. CONCLUSIONS: During scopolamine application as well as after intraglandular BoNT injections, a clinically relevant reduction in drooling was achieved in children with CP, demonstrating maximum effect 2 to 8 weeks after injections. This is the first controlled clinical trial that confirmed a significant effect of BoNT injections in the treatment of drooling. General anesthesia was needed for all children. BoNT injections show fewer and less serious side effects than transdermal scopolamine treatment. PMID- 15342831 TI - Continuous quality improvement: reducing unplanned extubations in a pediatric intensive care unit. AB - OBJECTIVE: Unplanned extubation (UEX) is a potentially serious complication of mechanical ventilation. Limited information is available regarding factors that contribute to UEXs and subsequent reintubation of children. We monitored UEXs in our pediatric intensive care unit (PICU) for a 5-year period to assess the incidence and patient conditions associated with UEX and to evaluate whether targeted interventions were associated with a reduced rate of UEXs. METHODS: Over a 5-year period, demographic and clinical information was collected prospectively on all patients who required an artificial airway while admitted to the PICU. Additional information was collected for patients who experienced an UEX. Educational sessions and care management protocols were developed, implemented, and modified according to issues identified via the monitoring program. RESULTS: From a total of 2192 patients who required 13 630 airway days (AWD), 141 (6%) patients experienced 164 UEXs. The overall rate of UEX for the study period was 1.2 UEXs per 100 AWD, and this rate decreased from 1.5 in the first year to 0.8 in the last year. UEXs were more common in children who were younger than 5 years (1.6 vs 0.6 UEX per 100 AWD) compared with older children. The UEX children experienced significantly longer length of mechanical ventilation (6 vs 3 days) and longer length of PICU stay (8 vs 4 days) compared with non-UEX children. Forty-six percent of the UEXs occurred in patients who were weaning from mechanical ventilation, and 22% of those patients required reintubation. CONCLUSIONS: We conclude that UEX in pediatric patients is associated with longer length of mechanical ventilation and length of stay in the PICU. A continuous quality improvement monitoring and educational program that identified high-risk patients for UEX (younger patients) and patients who were at low risk for subsequent reintubation (weaning patients) contributed to a reduction of these potentially adverse events. PMID- 15342833 TI - Ascertainment bias in Turner syndrome: new insights from girls who were diagnosed incidentally in prenatal life. AB - OBJECTIVE: To evaluate differences in phenotype and other clinical features between patients who have Turner syndrome diagnosed incidentally (on the basis of a prenatal karyotype performed for reasons unrelated to suspicion of Turner syndrome, eg, advanced maternal age) or traditionally (on the basis of either a prenatal karyotype performed for abnormal ultrasound findings or a postnatal karyotype performed for clinical findings suggesting Turner syndrome). METHODS: Analysis was performed on baseline data from 88 girls, aged 9 months to 4 years, who were randomized into a multicenter growth hormone intervention trial. Baseline information included a detailed medical history (especially of cardiac and renal anomalies), examination for presence of phenotypic features characteristic of Turner syndrome, length or height (depending on age) expressed as a standard deviation score, and weight standard deviation score. Patients were classified to either the "incidental" (N = 16) or the "traditional" (N = 72) diagnosis group as described above. RESULTS: The incidental group had significantly fewer total phenotypic features of Turner syndrome than the traditional group (3.6 +/- 2.9 vs 6.7 +/- 2.9). When subgrouped by karyotype, the proportion of patients who manifested phenotypic features was greatest in patients with a 45,X nonmosaic karyotype, lowest in the patients with 45,X/46,XX mosaicism, and intermediate in those with "other" karyotypes. Fewer congenital cardiac defects were observed in the incidental group (31%) compared with the traditional group (64%), but there was no difference between the groups in the prevalence of renal defects. Karyotype distribution differed significantly between the traditional and incidental groups: 74% versus 19% had the nonmosaic 45,X karyotype in the traditional group and 7% versus 56% had the mosaic 45,X/46,XX karyotype. CONCLUSIONS: Patients whose Turner syndrome was diagnosed incidentally had significantly fewer phenotypic features and cardiac defects, as well as a greater proportion of mosaic karyotypes, compared with patients whose Turner syndrome was diagnosed clinically. These results support the theory that significant ascertainment bias exists in our understanding Turner syndrome, with important implications for prenatal counseling. PMID- 15342832 TI - A population-based comparison of clinical and outcome characteristics of young children with serious inflicted and noninflicted traumatic brain injury. AB - OBJECTIVE: Diagnosing inflicted traumatic brain injury (TBI) in young children is difficult in practice. Comparisons of children with inflicted and noninflicted TBI may help to identify markers of inflicted TBI. The objective of this study was to compare inflicted and noninflicted TBI in terms of presenting complaints, clinical features, and hospital outcomes. METHODS: The presenting complaint, clinical finding, hospital course, and outcome of all children who were aged 2 years or younger in North Carolina and were admitted to a pediatric intensive care unit or died with a TBI in 2000 and 2001 were reviewed. Clinical presentation and injury types were compared between children with inflicted and noninflicted TBI. Risk ratios were used to compare clinical and outcome characteristics between the 2 groups. Among survivors, multivariate binomial regression was used to examine the adjusted risk of a poor outcome dependent on injury type. RESULTS: A total of 80 (52.6%) children had inflicted and 72 (47.3%) children had noninflicted TBI. Children with noninflicted TBI (not in a motor vehicle crash) were more likely to present to the emergency department asymptomatic (44.8% vs 8.3%) and to have a specific history of trauma than children with inflicted TBI. Retinal hemorrhage, metaphyseal fracture, rib fracture, and subdural hemorrhage were more commonly found in children with inflicted compared with noninflicted TBI. Skeletal survey and ophthalmologic examination combined would have missed 8 (10.0%) inflicted TBI cases. CONCLUSIONS: Manner of presentation and injury types are helpful in distinguishing inflicted TBI. Clinicians should not rule out inflicted TBI on the basis of skeletal survey and ophthalmoscopy alone but should proceed to computed tomography and/or magnetic resonance imaging. PMID- 15342834 TI - Longitudinal assessment of adaptive behavior in infants and young children with newly diagnosed epilepsy: influences of etiology, syndrome, and seizure control. AB - OBJECTIVES: The outcomes of childhood-onset epilepsy are highly varied and have several potential determinants. We examined the independent effects of syndrome type, seizure control, and etiology over time on adaptive behavior as measured by the Vineland Scales of Adaptive Behavior. METHODS: As part of a prospective community-based study of newly diagnosed epilepsy, parents of children who were younger than 3 years at the time of initial onset of epilepsy completed the Vineland Adaptive Behavior Scales screener version at entry into the study and once a year thereafter for up to 3 years. Longitudinal analyses were performed on the composite score as the primary outcome and on the 4 domain scores (communication, socialization, motor, and daily living) as confirmatory/secondary outcomes to determine the effects of syndrome (epileptic encephalopathy or other), seizure control (intractable or not), and etiology (symptomatic or not) on adaptive behavior at the time of initial diagnosis and over time. RESULTS: A total of 613 children were enrolled in the study, and 191 (31%) of these children met the age criterion for this analysis. Of these, 172 (90%) had adequate follow up and had completed baseline and at least 1 subsequent Vineland assessment. Overall, Vineland scores (composite and individual domains) were somewhat below average at baseline (initial diagnosis). All declined significantly over time. All of the effects at baseline, however, were limited to children with epileptic encephalopathies and symptomatic etiology. Substantial declines over time occurred in these children, and there was an independent effect of intractable seizures as well. In children with none of these factors ( approximately 75% of the study group), baseline scores were consistent with average performance for the test norms and there was no evidence of any decline over time. CONCLUSIONS: Children with an underlying symptomatic etiology or a syndrome that can be characterized as 1 of the epileptic encephalopathies demonstrate impaired adaptive behavior at the time of initial diagnosis and experience additional declines in assessments of age-adjusted performance over time. Our results suggest that future seizure outcome is not strongly reflected in adaptive behavior at initial diagnosis but that it takes its toll on the child over time. Understanding how each of these factors affects development and how they interact with each other is the next step in designing effective interventions for lessening the impact of these disorders on the child. In the majority of children with onset of epilepsy during infancy or early childhood, adaptive behavior is within the normal range and does not show any evidence of declining over time. Although this is encouraging, it does not contradict other studies that have demonstrated behavioral and relatively subtle cognitive difficulties in school aged children with epilepsy. Long-term follow-up in this cohort will permit us to examine the predictive value of the Vineland for later behavioral and cognitive difficulties in this group that, so far, seems to be doing well. PMID- 15342835 TI - Delivery room continuous positive airway pressure/positive end-expiratory pressure in extremely low birth weight infants: a feasibility trial. AB - OBJECTIVE: Although earlier studies have suggested that early continuous airway positive pressure (CPAP) may be beneficial in reducing ventilator dependence and subsequent chronic lung disease in the extremely low birth weight (ELBW) infant, the time of initiation of CPAP has varied, and there are no prospective studies of infants who have received CPAP or positive end-expiratory pressure (PEEP) from initial resuscitation in the delivery room (DR). Current practice for the ELBW infant includes early intubation and the administration of prophylactic surfactant, often in the DR. The feasibility of initiating CPAP in the DR and continuing this therapy without intubation for surfactant has never been determined prospectively in a population of ELBW infants. This study was designed to determine the feasibility of randomizing ELBW infants of <28 weeks' gestation to CPAP/PEEP or no CPAP/PEEP during resuscitation immediately after delivery, avoiding routine DR intubation for surfactant administration, initiating CPAP on neonatal intensive care unit (NICU) admission, and assessing compliance with subsequent intubation criteria. METHODS: Infants who were of <28 weeks' gestation, who were born in 5 National Institute of Child Health and Human Development Neonatal Research Network NICUs from July 2002 to January 2003, and for whom a decision had been made to provide full treatment after birth were randomized to receive either CPAP/PEEP or not using a neonatal T-piece resuscitator (NeoPuff). Infants would not be intubated for the sole purpose of surfactant administration in the DR. After admission to the NICU, all nonintubated infants were placed on CPAP and were to be intubated for surfactant administration only after meeting specific criteria: a fraction of inspired oxygen of >0.3 with an oxygen saturation by pulse oximeter of <90% and/or an arterial oxygen pressure of <45 mm Hg, an arterial partial pressure of carbon dioxide of >55 mm Hg, or apnea requiring bag and mask ventilation. RESULTS: A total of 104 infants were enrolled over a 6-month period: 55 CPAP and 49 control infants. No infant was intubated in the DR for the exclusive purpose of surfactant administration. Forty-seven infants were intubated for resuscitation in the DR: 27 of 55 CPAP infants and 20 of 49 control infants. Only 4 of the 43 infants who had a birth weight of <700 g and 3 of the 37 infants of <25 weeks' gestation were resuscitated successfully without positive pressure ventilation, and no difference was observed between the treatment groups. All infants of 23 weeks' gestation required intubation in the DR, irrespective of treatment group, whereas only 3 (14%) of 21 infants of 27 weeks' required such intubation. For infants who were not intubated in the DR, 36 infants (16 CPAP infants and 20 control infants) were subsequently intubated in the NICU by day 7, in accordance with the protocol. Overall, 80% of studied infants required intubation within the first 7 days of life. The care provided for 52 (95%) of 55 CPAP infants and 43 (88%) of the 49 control infants was in compliance with the study protocol, with an overall compliance of 91%. CONCLUSIONS: This study demonstrated that infants could be randomized successfully to a DR intervention of CPAP/PEEP compared with no CPAP/PEEP, with intubation provided only for resuscitation indications, and subsequent intubation for prespecified criteria. Forty-five percent (47 of 104) of infants <28 weeks' gestation required intubation for resuscitation in the DR. CPAP/PEEP in the DR did not affect the need for intubation at birth or during the subsequent week. Overall, 20% of infants did not need intubation by 7 days of life. This experience should be helpful in facilitating the design of subsequent prospective studies of ventilatory support in ELBW infants. PMID- 15342836 TI - Does sleep or sleep deprivation increase epileptiform discharges in pediatric electroencephalograms? AB - OBJECTIVE: Sleep deprivation before obtaining an electroencephalogram (EEG) is believed both to increase the likelihood of sleep during an EEG and to increase the detection of interictal epileptiform discharges. However, depriving a child of sleep poses a burden on both the parent and the child. The objective of this study was to compare the effects of sleep, standard sleep deprivation, partial sleep deprivation, and no sleep deprivation on the odds of an epileptiform abnormality in outpatient pediatric EEGs. METHODS: Data were collected from all pediatric EEGs performed at a busy, university-based neurologic practice during two 2-month periods. During the first period, all EEGs were performed as ordered, either standard sleep-deprived (SSD) or non-sleep-deprived (NSD). During the second 2 months, SSD EEGs were performed per routine. However, non-SSD families were instructed to keep their children awake 2 hours later the night before the EEG. Those who complied were classified as partially sleep-deprived (PSD). Patient characteristics across protocols were compared with chi(2) and analysis of variance tests as appropriate. The odds of epileptiform and abnormal findings associated with sleep, NSD, PSD, and SSD EEGs were calculated using logistic regression. RESULTS: Of 820 eligible EEGs, sleep occurred in 22% of NSD, 44% of PSD, and 57% of SSD EEGs. The sample size of this study allowed for an 85% power, with alpha of.05, to detect an absolute increased EEG yield of 10%. Neither the presence of sleep (odds ratio [OR]: 0.99; 95% confidence interval [CI]: 0.69 1.42) nor the use of PSD (OR: 0.90; 95% CI: 0.50-1.62) or SSD (OR: 0.96; 95% CI: 0.63-1.47) protocols increased the odds of epileptiform EEGs. CONCLUSIONS: Sleep deprivation should not be used routinely to increase the yield of pediatric EEGs. PMID- 15342837 TI - The EPIBEL study: outcomes to discharge from hospital for extremely preterm infants in Belgium. AB - OBJECTIVE: To determine mortality and morbidity at discharge from the hospital of a large population-based cohort of infants who were born at or=2500 g. Children who were born at 1000 to 1499 g (n = 839) generated 49% higher costs. Other birth conditions, independent of birth weight, were associated with higher kindergarten costs: family poverty (31%), congenital anomalies (29%), maternal education less than high school (20%), and no prenatal care (14%). Because of their prevalence, family poverty and low maternal education accounted for >75% of excess kindergarten costs. If 9% of infants who weighed between 1500 and 2499 g (n = 1027) could be delivered at 2500 g, then the state of Florida potentially could save 1 million dollars in kindergarten costs. Savings of a similar magnitude might be achieved if 3% of mothers who left school without a diploma (n = 1528) were to graduate. CONCLUSIONS: Any policy recommendation aimed at reducing education costs in kindergarten must take into consideration 3 factors: the prevalence of risk conditions whose amelioration is desired, the potential cost savings associated with reducing those conditions, and the costs of amelioration. Projecting these costs from information that is available at birth can assist school districts and state agencies in allocating resources. PMID- 15342846 TI - Use of incident reports by physicians and nurses to document medical errors in pediatric patients. AB - OBJECTIVES: To describe the proportion and types of medical errors that are stated to be reported via incident report systems by physicians and nurses who care for pediatric patients and to determine attitudes about potential interventions for increasing error reports. METHODS: A survey on use of incident reports to document medical errors was sent to a random sample of 200 physicians and nurses at a large children's hospital. Items on the survey included proportion of medical errors that were reported, reasons for underreporting medical errors, and attitudes about potential interventions for increasing error reports. In addition, the survey contained scenarios about hypothetical medical errors; the physicians and nurses were asked how likely they were to report each of the events described. Differences in use of incident reports for documenting medical errors between nurses and physicians were assessed with chi(2) tests. Logistic regression was used to determine the association between health care profession type and likelihood of reporting medical errors. RESULTS: A total of 140 surveys were returned, including 74 from physicians and 66 by nurses. Overall, 34.8% of respondents indicated that they had reported <20% of their perceived medical errors in the previous 12 months, and 32.6% had reported <40% of perceived errors committed by colleagues. After controlling for potentially confounding variables, nurses were significantly more likely to report >or=80% of their own medical errors than physicians (odds ratio: 2.8; 95% confidence interval: 1.3-6.0). Commonly listed reasons for underreporting included lack of certainty about what is considered an error (indicated by 40.7% of respondents) and concerns about implicating others (37%). Potential interventions that would lead to increased reporting included education about which errors should be reported (listed by 65.4% of respondents), feedback on a regular basis about the errors reported (63.8%) and about individual events (51.2%), evidence of system changes because of reports of errors (55.4%), and an electronic format for reports (44.9%). Although virtually all respondents would likely report a 10-fold overdose of morphine leading to respiratory depression in a child, only 31.7% would report an event in which a supply of breast milk is inadvertently connected to a venous catheter but is discovered before any breast milk goes into the catheter. CONCLUSIONS: Medical errors in pediatric patients are significantly underreported in incident report systems, particularly by physicians. Some types of errors are less likely to be reported than others. Information in incident reports is not a representative sample of errors committed in a children's hospital. Specific changes in the incident report system could lead to more reporting by physicians and nurses who care for pediatric patients. PMID- 15342847 TI - Prevalence, stability, and predictors of clinically significant behavior problems in low birth weight children at 3, 5, and 8 years of age. AB - OBJECTIVES: To examine the prevalence, stability, and predictors of clinically significant behavior problems in 869 preterm low birth weight (LBW) infants at 3, 5, and 8 years of age. METHODS: A prospective cohort study was conducted. Clinically significant behavior problems were assessed using dichotomized total problem Child Behavior Checklist scores in LBW children at ages 3, 5, and 8 years. Baseline sociodemographic and obstetric data were collected. Maternal General Health Questionnaire performed at 40 weeks' gestation was dichotomized at a score of 12 to give a measure of maternal psychological distress. Prevalence and stability of behavior problems at ages 3, 5, and 8 were determined and potential predictors of behavior problems at age 3, 5, and 8 were examined using multiple logistic regression. RESULTS: Prevalence of behavior problems remained at approximately 20% at 3, 5, and 8 years. Stability of behavior problems between different ages was approximately 50%. Significant behavior problems at ages 3, 5, and 8 were predicted by maternal psychological distress at 40 weeks (odds ratio [OR]: 1.59; 95% confidence interval [CI]: 1.21-2.09), maternal cigarette smoking during pregnancy (OR: 1.57; 95% CI: 1.20-2.04), Hispanic ethnicity (OR: 2.00; 95% CI: 1.24-3.24), and maternal age (OR: 0.97; 95% CI: 0.94-0.99). CONCLUSIONS: This sample had double the prevalence of behavior problems expected in the general child population. These problems showed stability over time. Cigarette smoking in pregnancy, maternal psychological distress at 40 weeks' gestation, maternal age, and Hispanic ethnicity all were significant predictors of the development of behavior problems from ages 3 to 8. These findings have implications for health policies on smoking and postnatal depression. PMID- 15342849 TI - Older age is a risk factor for the development of cardiovascular sequelae in Kawasaki disease. AB - OBJECTIVES: To clarify the characteristics of Kawasaki disease (KD) in children 6 years and older and to determine whether age is a risk factor for cardiovascular abnormalities. METHODS: Patients who had KD and were reported between 1999 and 2000 in the 16th nationwide survey of KD in Japan (n = 15,314) were analyzed. Patients who were aged 6 years or older (older group) were matched with patients who were aged 6 months to 3 years and were treated at the same hospital (younger groups). The total number of analyzed patients was 1498 (749 matched pairs). RESULTS: The proportion of complete KD in the older group was similar to that in the younger group. Recurrent cases in the older group were significantly more common than those in the younger group (9% vs 2%). The proportion of patients who were treated with intravenous gamma-globulin in the older group was significantly lower than that in the younger group (82% vs 87%). The proportion of older group patients who were treated with intravenous gamma-globulin at or after 7 days of illness was significantly higher than that in the younger group (35% vs 14%). There was a higher prevalence of cardiovascular abnormalities in the older group than in the younger group (20% vs 15%). Multivariate logistic regression analysis showed that older age was an independent risk factor for cardiovascular sequelae (odds ratio: 1.58; 95% confidence interval: 1.01-2.46). CONCLUSIONS: In children older than 6 years, age is an independent risk factor for cardiovascular sequelae in KD. PMID- 15342848 TI - Height and social adjustment: are extremes a cause for concern and action? AB - OBJECTIVE: Growth hormone treatment of hormone-sufficient short youths is predicated on the belief that short stature is associated with social problems with peers. This study assessed peer relations and social adjustment as a function of height in a community sample. METHODS: A cross-sectional study was conducted at 1 public school district in Western New York of students of both genders who attended grades 6 to 12 (N = 956). Target groups included participants of short stature (height or=1.6 standard deviation; >or=95th percentile; n = 58) and classmates of average height (between the 25th and 75th percentiles; n = 123) used for within-class comparisons. All remaining unclassified classmates (n = 704) also provided data. Self- and peer-rated assessment of social reputation and social acceptance was measured. RESULTS: Minimal effects of height on measures of social functioning were detected despite substantial statistical power. We detected no significant relationships between height and measures of friendship, popularity, or reputation with peers. Findings did not vary by gender of participant, by peer or self-report, or by whether data from the entire sample were used or target groups were contrasted with comparison participants. Shorter students were perceived by peers to be younger than their age. This influence was restricted to lower grades and did not have an impact on measures of social acceptance or reputation with peers. CONCLUSIONS: Extremes of stature in the general population either short or tall-have minimal detectable impact on peer perceptions of social behavior, friendship, or acceptance. Findings are not supportive of the need to intervene a priori because of the potential risk of negative stature-related social sequelae. Furthermore, if problems with peer relationships are identified among short or tall youths, then factors other than stature should be considered as etiologically important. PMID- 15342850 TI - Population-based impact of pneumococcal conjugate vaccine in young children. AB - OBJECTIVE: To determine the population impact of pneumococcal conjugate vaccine (PCV) on pneumococcal-related diseases, including pneumonia and otitis media. METHODS: Using administrative data from Tennessee Medicaid and 3 commercial insurance plans in upstate New York, we measured annual rates of medical visits for pneumococcal-related diseases (pneumococcal and nonspecific pneumonia and invasive disease; otitis media) and pneumococcal-unrelated diseases (other acute respiratory illnesses). Disease rates before (1995-2000 in Tennessee; 1998-2000 in New York) and after (2000-2002) PCV licensure were calculated for children aged <2 years (eligible for PCV) and those 3 to 5 years (not routinely given PCV). Because annual variations should affect both age groups similarly and vaccine-related outcomes should preferentially decline in younger children, ratios (<2:3-5 years) of disease rates before and after PCV licensure were compared. Expected disease rates were calculated for children aged <2 years in each postvaccine year. The difference between observed and expected disease rates was the estimated vaccine effect. RESULTS: In 2001-2002, there were 67 380 and 9485 child-years of observation for Tennessee and New York children aged <2 years, respectively. We observed fewer visits for pneumonia and invasive disease per 1000 children than expected in both regions: 20 fewer emergency department or outpatient visits in Tennessee and 33 fewer outpatient visits in New York. Otitis media visits declined by 118 and 430 per 1000 children in Tennessee and New York, respectively. CONCLUSIONS: Adding PCV to the childhood immunization schedule was associated with a 10-fold greater reduction in pneumonia and a 100-fold greater reduction in otitis media than the previously reported reduction in culture confirmed invasive pneumococcal diseases of 1.3 episodes per 1000 children aged <2 years. Although additional studies are needed to confirm the impact of routine immunization with PCV on pneumococcal-related disease, these results suggest that its impact is substantially greater than the effects on invasive disease alone. PMID- 15342851 TI - Duration of mechanical ventilation in life-threatening pediatric asthma: description of an acute asphyxial subgroup. AB - OBJECTIVE: Acute asphyxial asthma (AAA) is well described in adult patients and is characterized by a sudden onset that may rapidly progress to a near-arrest state. Despite the initial severity of AAA, mechanical ventilation often restores gas exchange promptly, resulting in shorter durations of ventilation. We believe that AAA can occur in children and can lead to respiratory failure that requires mechanical ventilation. Furthermore, children with rapid-onset respiratory failure that requires intubation in the emergency department (ED) are more likely to have AAA and a shorter duration of mechanical ventilation than those intubated in the pediatric intensive care unit (PICU). METHODS: An 11-year retrospective chart review (1991-2002) was conducted of all children who were aged 2 through 18 years and had the primary diagnosis of status asthmaticus and required mechanical ventilation. RESULTS: During the study period, 33 (11.4%) of 290 PICU admissions for status asthmaticus required mechanical ventilation. Thirteen children presented with rapid respiratory failure en route, on arrival, or within 30 minutes of arrival to the ED versus 20 children who progressed to respiratory failure later in their ED course or in the PICU. Mean duration of mechanical ventilation was significantly shorter in the children who presented with rapid respiratory failure versus those with progressive respiratory failure (29 +/- 43 hours vs 88 +/- 72 hours). Children with rapid respiratory failure had greater improvements in ventilation and oxygenation than those with progressive respiratory failure as measured by pre- and postintubation changes in arterial carbon dioxide pressure, arterial oxygen pressure/fraction of inspired oxygen ratio, and alveolar-arterial gradient. According to site of intubation, 23 children required intubation in the ED, whereas 10 were intubated later in the PICU. Mean duration of mechanical ventilation was significantly shorter in the ED group versus the PICU group (42 +/- 63 hours vs 118 +/- 46 hours). There were significantly greater improvements in ventilation and oxygenation in the ED group versus the PICU group as measured by pre- and postintubation changes in arterial carbon dioxide pressure and arterial oxygen pressure/fraction of inspired oxygen ratio. CONCLUSIONS: AAA occurs in children and shares characteristics seen in adult counterparts. Need for early intubation is a marker for AAA and may not represent a failure to maximize preintubation therapies. AAA represents a distinct form of life-threatening asthma and requires additional study in children. PMID- 15342852 TI - Daytime sleepiness and hyperactivity in children with suspected sleep-disordered breathing. AB - OBJECTIVES: Excessive daytime sleepiness (EDS) is seen less frequently as a presenting complaint in children with sleep-disordered breathing than in adults. Instead, symptoms of hyperactivity are often described. We hypothesized that children with suspected sleep-disordered breathing (S-SDB) were both sleepier and more hyperactive than control subjects. Furthermore, we hypothesized that overnight polysomnographic parameters correlated with sleepiness and hyperactivity. METHODS: A cross-sectional study was conducted at a university affiliated hospital and a community-based pediatric clinic. A total of 108 patients with S-SDB (mean [standard deviation] age: 7 +/- 4 years) and 72 control subjects (8 +/- 4 years) were recruited. A modified Epworth Sleepiness Scale (ESS) and the Conners Abbreviated Symptom Questionnaire were administered. Polysomnography was performed in patients with S-SDB. RESULTS: Patients with S SDB had a higher ESS (8.1 +/- 4.9 vs 5.3 +/- 3.9) and a higher Conners score (12.8 +/- 7.6 vs 9.0 +/- 6.2) than control subjects. On the basis of adult criteria, 28% of patients had EDS. There was no difference in the ESS and Conners scores of patients with primary snoring and patients with obstructive sleep apnea. The ESS had weak correlations with polysomnographic parameters. CONCLUSIONS: Although the ESS score of children with S-SDB was within the normal range for adults, these children were sleepier and more hyperactive than control subjects. However, these data should be confirmed by a population-based study. PMID- 15342853 TI - Exception from informed consent for pediatric resuscitation research: community consultation for a trial of brain cooling after in-hospital cardiac arrest. AB - OBJECTIVES: When prospective informed consent is not feasible, clinical research that presents more than minimal risk can proceed only after a community consultation and public disclosure process and the granting of exception from informed consent from the federal government. The applicability of exception from informed consent to pediatric resuscitation research has not been described. The objectives of this study were 1) to perform a community consultation and public disclosure process specific to a trial of induced hypothermia immediately after pediatric cardiac arrest and 2) to determine the applicability of exception from informed consent to randomized, controlled trials of emergency interventions after resuscitation from inpatient pediatric cardiac arrest. METHODS: Focus groups, information sheets with options for written responses, posted notices, e mails, and telephone conversations with parents of critically ill children and hospital staff were conducted at a tertiary care children's hospital. Data were stored, organized, and retrieved using NVivo qualitative analysis software (QSR International). RESULTS: In focus groups (n = 8), parents (n = 23) and hospital staff (n = 33) concluded that prospective informed consent is not feasible for a trial of induced hypothermia after inpatient pediatric cardiac arrest. Focus group participants endorsed exception from informed consent for a trial of induced hypothermia but only if study information is easily available prospectively and if all parents have an explicit opportunity to decline participation in a verbal conversation before study enrollment. Separate from and without knowledge of the focus group results, 7 (100%) of 7 parents of past or current patients and 21 (50%) of 42 hospital staff who provided written opinions endorsed exception from informed consent for this study. Five (12%) of 42 hospital staff opposed, and 16 (38%) of 42 were neutral. In telephone conversations, 14 (70%) of 20 parents of children who were previously resuscitated from cardiac arrest endorsed exception from informed consent for this study, 3 (15%) of 20 opposed, and 3 (15%) of 20 were unsure. CONCLUSIONS: Community consultation for inpatient resuscitation research can be conducted in a children's hospital, with hospital staff and parents of patients as the relevant community. Exception from informed consent is necessary and appropriate for a randomized trial of induced hypothermia begun within 30 minutes after pediatric cardiac arrest. A process in which families are informed prospectively and have a pre-enrollment option to decline participation will likely be acceptable to families, health care providers, and the institution. PMID- 15342854 TI - Intussusception in early childhood: a cohort study of 1.7 million children. AB - OBJECTIVE: To describe incidence and temporal trends of intussusceptions in Danish children during 1980 to 2001. METHODS: A population-based cohort study was conducted of 1.67 million children who were younger than 5 years during 1980 to 2001 and were followed up for 6.66 million person-years. The Danish National Patient Registry was used to identify cases of intussusception in the cohort. Age specific incidence rates were main outcome measure. RESULTS: A total of 1814 cases of intussusception among children who were younger than 5 years were reported from 1980 to 2001. The incidence rate remained fairly constant during 1980 to 1990 but decreased by 55% (95% confidence interval: 43%-65%) from 1990 to 2001. The reduction was most pronounced among children aged 3 to 5 months. CONCLUSIONS: The incidence of intussusception among Danish children declined significantly during the 1990s, particularly among infants 3 to 5 months of age. PMID- 15342855 TI - Decline in varicella-related hospitalizations and expenditures for children and adults after introduction of varicella vaccine in the United States. AB - OBJECTIVE: Universal childhood immunization against varicella in the United States, first recommended in 1995, was predicted to lead to significant decreases in varicella-related hospitalization rates and corresponding charges. Previous studies have not found such effects. We studied trends in varicella-related hospitalization rates and associated charges before and after introduction of varicella vaccine. METHODS: We examined hospitalization and charge data from the Nationwide Inpatient Sample for the years 1993-2001, representative of national hospitalization patterns for children and adults. We derived weighted estimates of population-adjusted, varicella-related hospitalization rates and inflation adjusted, varicella-related hospital charges. RESULTS: The annual varicella related hospitalization rate exceeded 0.5 hospitalizations per 10,000 US population from 1993 to 1995, declined to 0.26 per 10,000 by 1999, and again halved to 0.13 per 10,000 by 2001. Hospitalization rates declined most substantially among individuals primarily targeted for vaccination (0- to 4-year old children) but decreased among youths aged 5 to 19 years and among adults as well. Concomitantly, varicella-related hospital charges declined from 161.1 million dollars (95% confidence interval: 130.5 million dollars-191.8 million dollars) in 1993 to 66.3 million dollars (95% confidence interval: 50.9 million dollars-81.7 million dollars) in 2001 (all 2001 US dollars). Among expected primary payers, inflation-adjusted declines in varicella-related hospital discharges-related charges accrued to Medicaid, private insurance, and "other" payers (including uninsured and self-pay) but not to Medicare. CONCLUSIONS: This national analysis indicates a clinically and statistically significant reduction in varicella-related hospitalizations for children and adults associated with childhood varicella immunization in the United States and a corresponding significant decrease in hospital charges. PMID- 15342856 TI - Thimerosal-containing vaccines and autistic spectrum disorder: a critical review of published original data. AB - OBJECTIVE: The issue of thimerosal-containing vaccines as a possible cause of autistic spectrum disorders (ASD) and neurodevelopmental disorders (NDDs) has been a controversial topic since 1999. Although most practitioners are familiar with the controversy, many are not familiar with the type or quality of evidence in published articles that have addressed this issue. To assess the quality of evidence assessing a potential association between thimerosal-containing vaccines and autism and evaluate whether that evidence suggests accepting or rejecting the hypothesis, we systematically reviewed published articles that report original data pertinent to the potential association between thimerosal-containing vaccines and ASD/NDDs. METHODS: Articles for analysis were identified in the National Library of Medicine's Medline database using a PubMed search of the English-language literature for articles published between 1966 and 2004, using keywords thimerosal, thiomersal, mercury, methylmercury, or ethylmercury alone and combined with keywords autistic disorder, autistic spectrum disorder, and neurodevelopment. In addition, we used the "related links" option in PubMed and reviewed the reference sections in the identified articles. All original articles that evaluated an association between thimerosal-containing vaccines and ASD/NDDs or pharmacokinetics of ethylmercury in vaccines were included. RESULTS: Twelve publications that met the selection criteria were identified by the literature search: 10 epidemiologic studies and 2 pharmacokinetic studies of ethylmercury. The design and quality of the studies showed significant variation. The preponderance of epidemiologic evidence does not support an association between thimerosal-containing vaccines and ASD. Epidemiologic studies that support an association are of poor quality and cannot be interpreted. Pharmacokinetic studies suggest that the half-life of ethylmercury is significantly shorter when compared with methylmercury. CONCLUSIONS: Studies do not demonstrate a link between thimerosal-containing vaccines and ASD, and the pharmacokinetics of ethylmercury make such an association less likely. Epidemiologic studies that support a link demonstrated significant design flaws that invalidate their conclusions. Evidence does not support a change in the standard of practice with regard to administration of thimerosal-containing vaccines in areas of the world where they are used. PMID- 15342857 TI - The effect of chronic or intermittent hypoxia on cognition in childhood: a review of the evidence. AB - OBJECTIVE: A review of the evidence concerning the effect of chronic or intermittent hypoxia on cognition in childhood was performed by using both a systematic review of the literature and critical appraisal criteria of causality. Because of the significant impact of behavioral disorders such as attention deficit/hyperactivity disorder on certain cognitive functions as well as academic achievement, the review also included articles that addressed behavioral outcomes. METHODS: Both direct and indirect evidence were collected. A structured Medline search was conducted from the years 1966-2000 by using the OVID interface. Both English- and non-English-language citations were included. Significant articles identified by the reviewers up to 2003 were also included. To be included as direct evidence, an article needed to be an original report in a peer-reviewed journal with data on cognitive, behavioral, or academic outcomes in children up to 14 years old, with clinical conditions likely to be associated with exposure to chronic or intermittent hypoxia. Indirect evidence from other reviews and publications in closely related fields, including experimental studies in adults, was used to help formulate conclusions. Two reviewers screened abstracts and titles. Each article included as direct evidence received a structured evaluation by 2 reviewers. Adjudication of differences was performed by a group of 2 reviewers and a research consultant. After this review, tables of evidence were constructed that were used as the basis for group discussion and consensus development. Indirect evidence assigned by topic to specific reviewers was also presented as part of this process. A formal procedure was used to rank the studies by design strength. The critical appraisal criteria for causation described in Evidence Based Pediatrics and Child Health (Moyer V, Elliott E, Davis R, et al, eds. London, United Kingdom: BMJ Books; 2000:46-55) were used to develop consensus on causality. RESULTS: A total of 788 literature citations were screened. For the final analysis, 55 articles met the criteria for inclusion in the direct evidence. Of these, 43 (78.2%) reported an adverse effect. Of the 37 controlled studies, 31 (83.8%) reported an adverse effect. Adverse effects were noted at every level of arterial oxygen saturation and for exposure at every age level except for premature newborns. The studies were classified into 5 clinical categories: congenital heart disease (CHD), sleep-disordered breathing (SDB), asthma, chronic ventilatory impairment, and respiratory instability in infants. Two of these categories, CHD and SDB, which accounted for 42 (76.4%) of the included articles, fulfilled the Evidence Based Pediatrics and Child Health criteria for causation. The indirect evidence included 8 reviews, 1 meta analysis, and 10 original reports covering the fields of adult anoxia, animal research, SDB in adults, natural and experimental high-altitude studies, perinatal hypoxic-ischemic encephalopathy, anemia, and carbon-monoxide poisoning. The studies of high-altitude and carbon-monoxide poisoning provided evidence for causality. CONCLUSIONS: Adverse impacts of chronic or intermittent hypoxia on development, behavior, and academic achievement have been reported in many well designed and controlled studies in children with CHD and SDB as well as in a variety of experimental studies in adults. This should be taken into account in any situation that may expose children to hypoxia. Because adverse effects have been noted at even mild levels of oxygen desaturation, future research should include precisely defined data on exposure to all levels of desaturation. PMID- 15342858 TI - The Internet--a new medical problem or invaluable ally? AB - The public's increasing use of the Internet as a personal medical consultant poses new problems and special opportunities for the practicing physician. This article outlines how the Internet is already changing medical practice and how physicians can begin to use this resource to an advantage in their daily medical practice. PMID- 15342859 TI - Healthy Steps: a case study of innovation in pediatric practice. AB - Healthy Steps (HS) represents a significant innovation in the way pediatric primary care can be delivered. Based on the standards and principles of Bright Futures and the American Academy of Pediatrics Health Supervision Guidelines, HS enhances and expands traditional pediatric care by including a child development specialist (Healthy Steps specialist) as part of the pediatric practice team. Services offered by this person, typically a nurse, early childhood educator, or social worker, include more time to spend discussing preventive issues during well-child visits, home visits, a telephone information line exclusively addressing developmental and behavioral concerns, new written materials, and more seamless linkages to community resources and parent support groups. The original HS cohort consisted of 15 pediatric practices in a variety of settings (private practices, health centers, pediatric training programs). Evaluated for the effects of HS on their family were 3737 intervention and comparison families. HS families received significantly more preventive and developmental services, compared to families in the control group. HS families were also less likely to be dissatisfied with their pediatric primary care. Additionally, HS had a positive impact on parenting in many areas including adherence to health visits, nutritional practices, developmental stimulation, appropriate disciplinary techniques, and correct sleeping position. Other outcome measures (such as initiation or duration of breastfeeding, child development knowledge, sense of competence, and reports of child language development at 2 years of age) did not differ between intervention and comparison group. Compared to other early childhood intervention efforts, HS offers a comparable positive impact on parenting at a relatively inexpensive cost: an estimated 400 dollars per family per year (compared to 4500 dollars from Early Head Start). Approximately 3 years after the evaluation of HS ended, 10 of the original 24 sites are still in operation, and an additional 24 sites have started up. Although funding and reimbursement remain an important barrier, continued growth of HS suggests an abiding interest in this approach to expand and enhance preventive and developmental care in pediatric primary care. PMID- 15342860 TI - 2003 C. Anderson Aldrich award lecture: enhancing developmental services in child health supervision--an idea whose time has truly arrived. PMID- 15342861 TI - The state of pediatrics residency training: a period of transformation of graduate medical education. AB - Graduate medical education is in a period of transformation. This article reviews the state of pediatrics residency training by summarizing the changing demographics within training programs, examining the new educational paradigm with an emphasis on competency-based education and continuous professional development, and describing forces influencing the workplace and the focus on work-life balance. Strategies are suggested for leaders in graduate medical education to meet the challenges experienced during this period of transformation. PMID- 15342862 TI - Preschoolers, computers, and school readiness: are we on to something? PMID- 15342863 TI - Palliative care: a supportive adjunct to pediatric phase I clinical trials for anticancer agents? PMID- 15342864 TI - Water births: a naked emperor. PMID- 15342865 TI - Head Start's National Reporting System: a work in progress. PMID- 15342866 TI - Resuscitation research and the final rule: is there an impasse? PMID- 15342867 TI - Nontherapeutic use of antimicrobial agents in animal agriculture: implications for pediatrics. AB - Antimicrobial resistance is widespread. Overuse or misuse of antimicrobial agents in veterinary and human medicine is responsible for increasing the crisis of resistance to antimicrobial agents. The American Academy of Pediatrics, in conjunction with the US Public Health Service, has begun to address this problem by disseminating policies on the judicious use of antimicrobial agents in humans. Between 40% and 80% of the antimicrobial agents used in the United States each year are used in food animals; many are identical or very similar to drugs used in humans. Most of this use involves the addition of low doses of antimicrobial agents to the feed of healthy animals over prolonged periods to promote growth and increase feed efficiency or at a range of doses to prevent disease. These nontherapeutic uses contribute to resistance and create health dangers for humans. This report will describe how antimicrobial agents are used in animal agriculture and review the mechanisms by which such uses contribute to resistance in human pathogens. Although therapeutic use of antimicrobial agents in agriculture clearly contributes to the development of resistance, this report will concentrate on nontherapeutic uses in healthy animals. PMID- 15342868 TI - Dealing with the parent whose judgment is impaired by alcohol or drugs: legal and ethical considerations. AB - An estimated 11 to 17.5 million children are being raised by a substance-abusing parent or guardian. The importance of this statistic is undeniable, particularly when a patient is brought to a pediatric office by a parent or guardian exhibiting symptoms of judgment impairment. Although the physician-patient relationship exists between the pediatrician and the minor patient, other obligations (some perceived and some real) should be considered as well. In managing encounters with impaired parents who may become disruptive or dangerous, pediatricians should be aware of their responsibilities before acting. In addition to fulfilling the duty involved with an established physician-patient relationship, the pediatrician should take reasonable care to safeguard patient confidentiality; protect the safety of the patient and other patients, visitors, and employees; and comply with reporting mandates. This clinical report identifies and discusses the legal and ethical concepts related to these circumstances. The report offers implementation suggestions when establishing anticipatory office procedures and training programs for staff on what to do (and not do) in such situations to maximize the patient's well-being and safety and minimize the liability of the pediatrician. PMID- 15342869 TI - Classifying recommendations for clinical practice guidelines. AB - Clinical practice guidelines are intended to improve the quality of clinical care by reducing inappropriate variations, producing optimal outcomes for patients, minimizing harm, and promoting cost-effective practices. This statement proposes an explicit classification of recommendations for clinical practice guidelines of the American Academy of Pediatrics (AAP) to promote communication among guideline developers, implementers, and other users of guideline knowledge, to improve consistency, and to facilitate user understanding. The statement describes 3 sequential activities in developing evidence-based clinical practice guidelines and related policies: 1) determination of the aggregate evidence quality in support of a proposed recommendation; 2) evaluation of the anticipated balance between benefits and harms when the recommendation is carried out; and 3) designation of recommendation strength. An individual policy can be reported as a "strong recommendation," "recommendation," "option," or "no recommendation." Use of this classification is intended to improve consistency and increase the transparency of the guideline-development process, facilitate understanding of AAP clinical practice guidelines, and enhance both the utility and credibility of AAP clinical practice guidelines. PMID- 15342870 TI - Overcrowding crisis in our nation's emergency departments: is our safety net unraveling? AB - Emergency departments (EDs) are a vital component in our health care safety net, available 24 hours a day, 7 days a week, for all who require care. There has been a steady increase in the volume and acuity of patient visits to EDs, now with well over 100 million Americans (30 million children) receiving emergency care annually. This rise in ED utilization has effectively saturated the capacity of EDs and emergency medical services in many communities. The resulting phenomenon, commonly referred to as ED overcrowding, now threatens access to emergency services for those who need them the most. As managers of the pediatric medical home and advocates for children and optimal pediatric health care, there is a very important role for pediatricians and the American Academy of Pediatrics in guiding health policy decision-makers toward effective solutions that promote the medical home and timely access to emergency care. PMID- 15342871 TI - Prenatal screening and diagnosis for pediatricians. AB - The pediatrician who cares for a child with a birth defect or genetic disorder may be in the best position to alert the family to the possibility of a recurrence of the same or similar problems in future offspring. The family may wish to know about and may benefit from methods that convert probability statements about recurrence risks into more precise knowledge about a specific abnormality in the fetus. The pediatrician also may be called on to discuss abnormal prenatal test results as a way of understanding the risks and complications that the newborn infant may face. Along with the increase in knowledge brought about by the sequencing of the human genome, there has been an increase in the technical capabilities for diagnosing many chromosome abnormalities, genetic disorders, and isolated birth defects in the prenatal period. The purpose of this report is to update the pediatrician about indications for prenatal diagnosis, current techniques used for prenatal diagnosis, and the status of maternal screenings for detection of fetal abnormalities. PMID- 15342872 TI - Aggression, mania, and hypomania induction associated with atomoxetine. PMID- 15342873 TI - Delay in referral to early-intervention services. PMID- 15342874 TI - Compassion or opportunism? PMID- 15342875 TI - Compassion or opportunism? PMID- 15342876 TI - Where there is no shame, there is no honor (Ethiopian proverb). PMID- 15342877 TI - Military pediatricians in Southwest Asia. PMID- 15342878 TI - Socioeconomics and otitis media. PMID- 15342879 TI - Reduced bone mineralization in infants fed palm olein-containing formula: a randomized, double-blinded, prospective trial. PMID- 15342880 TI - The need for consistency in 407 reviews. PMID- 15342881 TI - Management of hyperbilirubinemia in newborns: measuring performance using a benchmarking model. PMID- 15342882 TI - Hyperbilirubinemia benchmarking. PMID- 15342883 TI - Yes, steroids are safe in infants with asthma-like symptoms. PMID- 15342884 TI - Missed opportunities for perinatal HIV prevention among HIV-exposed infants born 1996-2000, pediatric spectrum of HIV disease cohort. PMID- 15342885 TI - Rheumatic Fever in children under 5 years. PMID- 15342886 TI - A simple treatment for head lice: dry-on, suffocation-based pediculicide. AB - OBJECTIVES: The emergence of drug-resistant lice has created the need for new therapies. This study assesses a new method without neurotoxins, extensive household cleaning, or nit removal. METHODS: One hundred thirty-three subjects participated in 2 open clinical trials. In the first trial, 93 subjects completed treatment using a nontoxic, dry-on, suffocation-based, pediculicide lotion, minimal household cleaning measures, and physical removal of the nits. In the second trial, 40 subjects completed treatment using an identical protocol except that the nits were not removed. Head lice infestation was defined as a wet combing test showing lice. Cure was defined as a wet combing test showing no lice, with an absence of symptoms. Subjects were contacted 6 months later, for assessment of their disease status. RESULTS: Cure was achieved for 97% of the patients in the first trial and 95% in the second trial. Remission at the follow up assessment was reported for 77 of 82 subjects (94%) in the first trial and 36 of 38 subjects (95%) in the second trial. There was no statistical difference in cure rates or remission rates between the protocols with and without nit removal. The overall cure rate was 96%, with a remission rate of 94%. There were no adverse effects. CONCLUSIONS: Dry-on, suffocation-based, pediculicide lotion effectively treats head lice without neurotoxins, nit removal, or extensive house cleaning. These results are comparable or superior to the results previously reported for treatments with permethrin, pyrethrin, and malathion. PMID- 15342887 TI - Watching sex on television predicts adolescent initiation of sexual behavior. AB - BACKGROUND: Early sexual initiation is an important social and health issue. A recent survey suggested that most sexually experienced teens wish they had waited longer to have intercourse; other data indicate that unplanned pregnancies and sexually transmitted diseases are more common among those who begin sexual activity earlier. The American Academy of Pediatrics has suggested that portrayals of sex on entertainment television (TV) may contribute to precocious adolescent sex. Approximately two-thirds of TV programs contain sexual content. However, empirical data examining the relationships between exposure to sex on TV and adolescent sexual behaviors are rare and inadequate for addressing the issue of causal effects. DESIGN AND PARTICIPANTS: We conducted a national longitudinal survey of 1792 adolescents, 12 to 17 years of age. In baseline and 1-year follow up interviews, participants reported their TV viewing habits and sexual experience and responded to measures of more than a dozen factors known to be associated with adolescent sexual initiation. TV viewing data were combined with the results of a scientific analysis of TV sexual content to derive measures of exposure to sexual content, depictions of sexual risks or safety, and depictions of sexual behavior (versus talk about sex but no behavior). OUTCOME MEASURES: Initiation of intercourse and advancement in noncoital sexual activity level, during a 1-year period. RESULTS: Multivariate regression analysis indicated that adolescents who viewed more sexual content at baseline were more likely to initiate intercourse and progress to more advanced noncoital sexual activities during the subsequent year, controlling for respondent characteristics that might otherwise explain these relationships. The size of the adjusted intercourse effect was such that youths in the 90th percentile of TV sex viewing had a predicted probability of intercourse initiation that was approximately double that of youths in the 10th percentile, for all ages studied. Exposure to TV that included only talk about sex was associated with the same risks as exposure to TV that depicted sexual behavior. African American youths who watched more depictions of sexual risks or safety were less likely to initiate intercourse in the subsequent year. CONCLUSIONS: Watching sex on TV predicts and may hasten adolescent sexual initiation. Reducing the amount of sexual content in entertainment programming, reducing adolescent exposure to this content, or increasing references to and depictions of possible negative consequences of sexual activity could appreciably delay the initiation of coital and noncoital activities. Alternatively, parents may be able to reduce the effects of sexual content by watching TV with their teenaged children and discussing their own beliefs about sex and the behaviors portrayed. Pediatricians should encourage these family discussions. PMID- 15342888 TI - Use of voice recognition software in an outpatient pediatric specialty practice. AB - BACKGROUND: Voice recognition software (VRS), with specialized medical vocabulary, is being promoted to enhance physician efficiency, decrease costs, and improve patient safety. This study reports the experience of a pediatric subspecialist (pediatric gastroenterology) physician with the use of Dragon Naturally Speaking (version 6; ScanSoft Inc, Peabody, MA), incorporated for use with a proprietary electronic medical record, in a large university medical center ambulatory care service. METHODS: After 2 hours of group orientation and 2 hours of individual VRS instruction, the physician trained the software for 1 month (30 letters) during a hospital slowdown. Set-up, dictation, and correction times for the physician and medical transcriptionist were recorded for these training sessions, as well as for 42 subsequently dictated letters. Figures were extrapolated to the yearly clinic volume for the physician, to estimate costs (physician: 110 dollars per hour; transcriptionist: 11 dollars per hour, US dollars). RESULTS: The use of VRS required an additional 200% of physician dictation and correction time (9 minutes vs 3 minutes), compared with the use of electronic signatures for letters typed by an experienced transcriptionist and imported into the electronic medical record. When the cost of the license agreement and the costs of physician and transcriptionist time were included, the use of the software cost 100% more, for the amount of dictation performed annually by the physician. CONCLUSIONS: VRS is an intriguing technology. It holds the possibility of streamlining medical practice. However, the learning curve and accuracy of the tested version of the software limit broad physician acceptance at this time. PMID- 15342889 TI - Comparison of two programs to teach firearm injury prevention skills to 6- and 7 year-old children. AB - BACKGROUND: Each year, hundreds of children unintentionally kill or injure other children while playing with firearms in the United States. Although the numbers of these deaths and injuries are distressing, few prevention programs have been developed to prevent gun play among children. OBJECTIVE: This study compared the efficacy of 2 programs designed to prevent gun play among young children. DESIGN: A posttest-only, control group design with 2 treatment groups was used. Children were randomly assigned to 1 of 2 treatment groups or a control group. For all children who did not exhibit the skills after training, 1 in situ (ie, real-life situation) training session was conducted. SETTING: Participant recruitment, training sessions, and assessments were all conducted in the children's after school program facility. PARTICIPANTS: Forty-five children, 6 or 7 years of age, were recruited for participation. INTERVENTIONS: The efficacy of the Eddie Eagle GunSafe Program, developed by the National Rifle Association, and a behavioral skills training program that emphasized the use of instruction, modeling, rehearsal, and feedback was evaluated. MAIN OUTCOME MEASURES: The criterion firearm safety behaviors included both motor and verbal responses, which were assessed in a naturalistic setting and then assigned a numerical value based on a scale of 0 to 3. RESULTS: Both programs were effective for teaching children to verbalize the safety skills message (don't touch the gun, get away, and tell an adult). However, children who received behavioral skills training were significantly more likely to demonstrate the desired safety skills in role playing assessments and in situ assessments than were children who received Eddie Eagle program training. In addition, in situ training was found to be effective for teaching the desired safety skills for both groups of children. CONCLUSIONS: Injury prevention programs using education-based learning materials are less effective for teaching children the desired safety skills, compared with programs incorporating active learning approaches (eg, modeling, rehearsal, and feedback). The efficacy of both types of injury prevention programs for teaching the desired skills could be significantly enhanced with the use of in situ training. This program, when implemented with 6- and 7-year-old children, was effective in teaching the desired safety skills. PMID- 15342890 TI - Puberty and the onset of substance use and abuse. AB - OBJECTIVE: Substance abuse remains one of the major threats to adolescent health in Western cultures. The study aim was to ascertain the extent of association between pubertal development and early adolescent substance use. METHODS: The design was a cross-sectional survey of 10- to 15-year-old subjects in the states of Washington, United States, and Victoria, Australia. Participants were 5769 students in grades 5, 7, and 9, drawn as a 2-stage cluster sample in each state, and the questionnaire was completed in the school classrooms. The main outcomes of the study were lifetime substance use (tobacco use, having been drunk, or cannabis use), recent substance use (tobacco, alcohol, or cannabis use in the previous month), and substance abuse (daily smoking, any binge drinking, drinking at least weekly, or cannabis use at least weekly). RESULTS: The odds of lifetime substance use were almost twofold higher (odds ratio [OR]: 1.7; 95% confidence interval [CI]: 1.4-2.1) in midpuberty (Tanner stage III) and were threefold higher (OR: 3.1; 95% CI: 2.4-4.2) in late puberty (Tanner stage IV/V), after adjustment for age and school grade level. Recent substance use was moderately higher (OR: 1.4; 95% CI: 1.0-1.9) in midpuberty and more than twofold higher (OR: 2.3; 95% CI: 1.7-3.3) in late puberty. The odds of substance abuse were twofold higher (OR: 2.0; 95% CI: 1.2-3.2) in midpuberty and more than threefold higher (OR: 3.5; 95% CI: 2.2-5.4) in late puberty. Reporting most friends as substance users was more likely in the later stages of pubertal development, a relationship that accounted in part for the association found between later pubertal stage and substance abuse. CONCLUSIONS: Pubertal stage was associated with higher rates of substance use and abuse independent of age and school grade level. Early maturers had higher levels of substance use because they entered the risk period at an earlier point than did late maturers. The study findings support prevention strategies and policies that decrease recreational substance use within the peer social group in the early teens. PMID- 15342891 TI - Predictive value of a cross-cultural asthma case-detection tool in an elementary school population. AB - OBJECTIVE: Bronchial asthma, which affects approximately 5 million US children, is vastly underdiagnosed and treated, particularly among minorities and those of low socioeconomic status. Because current methods of detecting those at greatest risk of asthma in a multicultural setting appear inadequate, we assessed the validity and reliability of a new asthma questionnaire across 3 dominant cultures in Orange County, California (white, Hispanic, and Vietnamese). METHODS: Children in grades 1, 3, and 5 and their families, in 3 different schools representative of these major ethnic groups, were randomly selected to participate in the validation process. Two schools with low socioeconomic status and dominant Hispanic or Vietnamese minorities were designated inner-city schools, whereas the third school was a suburban school with predominately white students. Participants completed a 7-question, 11-element questionnaire in their primary language, followed by an asthma evaluation (history, physical examination, and spirometry) by an asthma specialist (who was blinded with respect to the results of the questionnaire), at their respective schools. The physician then made a determination regarding the presence and severity (according to National Institutes of Health guidelines) of asthma. Several weeks later, the entire student body was asked to complete the questionnaire at home and return it to school for analysis. Validation of each item was evaluated for sensitivity, specificity, and positive and negative predictive values, and application of univariate analyses provided an estimated probability of an asthma diagnosis by the asthma specialist. A "best-fit" algorithm was determined with all 11 elements, if possible, and an abbreviated algorithm that selected the fewest question combination that yielded the best asthma predictability was established. Reliability was established with the percent agreement between the 2 questionnaires and the kappa statistic. RESULTS: Of the 401 children/families who participated in the validation analysis, 45% were Hispanic, 22% white, 19% Vietnamese, and 15% other. The overall prevalence of asthma specialist-diagnosed asthma was 28%, with 65% of cases being graded as intermittent and 35% as persistent. Sixty-two percent of the children had not been previously diagnosed with asthma. There were no significant differences among cultures in sensitivity or specificity for any of the individual questions or the complete or abbreviated algorithms. The abbreviated algorithm with 3 questions, ie, question 1 (asthma in the past 2 years), question 4 (cough, chest tightness, trouble breathing, or wheezing with exercise), and question 6 (same symptoms in the morning or day in the past 4 weeks) yielded comparable sensitivity and specificity for the complete algorithm in all groups. The abbreviated algorithm had >86% predictability in detecting children with persistent asthma and 56% predictability in detecting children with intermittent asthma. Reliability was also excellent, with percent agreement usually > 80% and kappa values of >.70. CONCLUSIONS: This asthma detection tool has been shown to be suitable for detecting persistent asthma in a multicultural inner-city population, as well as in a suburban setting. An abbreviated algorithm with 3 questions and >80% predictability in detecting persistent asthma seems ideal for evaluating large numbers of school-aged children. The school setting is an excellent site for identifying children with asthma. Although there is concern that subjects detected in the school setting might not have access to ongoing medical care, case detection is an important first step that could lead to earlier diagnosis and treatment. Reducing the barriers to good care in inner-city environments is the next step. PMID- 15342892 TI - Hawaii's healthy start home visiting program: determinants and impact of rapid repeat birth. AB - OBJECTIVE: Healthy People 2010 calls for reductions in rapid repeat births (RRBs), defined as births occurring within 24 months after a previous birth for women of all ages, and prevention of repeat births during adolescence, regardless of the birth interval. Home visiting has been promoted as a mechanism to prevent child abuse and neglect and to improve pregnancy outcomes. This study aims to assess the impact of home visiting in preventing RRB and its malleable determinants and assesses the influence of RRB on the mother and the index child. We hypothesized that maternal desire to have a RRB, access to a family planning site, and use of birth control would be significant malleable determinants and that the effects of the program in preventing RRB would be mediated through its influence on these variables. We also hypothesized that the occurrence of RRB would result in increased stress and family dysfunction, resulting in adverse maternal and child outcomes such as severe maternal stress, maternal neglect of the index child, decreased maternal warmth toward the index child, and increased behavior problems of the index child. METHODS: The Healthy Start Program (HSP) is a home visiting program to prevent child abuse and neglect and to promote child health and development among newborns of families identified as being at risk for child maltreatment. This study was a randomized, controlled trial of Hawaii's HSP, in which eligible families were randomly assigned to home-visited and control groups. A total of 643 families at risk for child abuse were enrolled between November 1994 and December 1995. Data to measure RRB and malleable determinants were collected through structured maternal interviews and observation of the home environment. We measured RRB through maternal self-report by asking about a subsequent birth in follow-up interviews at 1, 2, and 3 years. To measure the malleable determinants, we measured the mother's desire for a RRB at baseline and at the 1-year interview and determined whether she had access to a family planning site. The mother was also asked which contraceptive methods she had ever used in the past and which methods, if any, she used in the year following the index child's birth. We measured 3 maternal parenting outcomes at the year 3 follow-up interview, ie, parenting stress, neglectful behavior toward the index child, and warmth toward the index child. We used odds ratios with 95% confidence intervals (CIs) to measure the strength of associations. Multiple logistic regression was used to assess 1) program effects on RRB and its malleable determinants, 2) the impact of the malleable determinants on RRB, and 3) the association between RRB and adverse maternal and child outcomes. RESULTS: Each year, 88% of the sample completed a follow-up interview; 81% completed all 3 follow-up interviews. There was no program impact on RRB for mothers overall (HSP: 21%; control: 20%; adjusted odds ratio [AOR]: 1.05; 95% CI: 0.69-1.58). HSP and control groups did not differ significantly in any of the malleable determinants of RRB. When we combined the 2 study groups, malleable determinants had significant effects on RRB. Mothers with a desire to have a child within 2 years after the index birth were significantly more likely to have a RRB, whether this desire was expressed at baseline (AOR: 2.48; 95% CI: 1.32-4.64) or at the year 1 interview (AOR: 2.86; 95% CI: 1.57-5.22). Lack of access to a family planning site at baseline was not associated with RRB, but there was a trend toward a greater likelihood of RRB among those lacking a site at 1 year (AOR: 1.61; 95% CI: 0.93-2.79). Women who had never used birth control before the index birth were more likely to have a RRB (AOR: 1.89; 95% CI: 1.20-2.98), and there was a trend toward a greater likelihood of RRB among women who did not use birth control in the year following the index child's birth (AOR: 1.67; 95% CI: 0.98 2.82). At the 3-year follow-up interview, mothers with a RRB were more likely to have adverse maternal and child outcomes. There was greater likelihood of severe maternal parenting stress (AOR: 2.29; 95% CI: 1.17-4.48), neglectful behavior toward the index child (AOR: 2.42; 95% CI: 1.41-4.18), and poor warmth toward the index child (AOR: 2.84; 95% CI: 1.71-4.42). In families with a RRB, the index child was more likely to exhibit internalizing behavior (AOR: 1.64; 95% CI: 1.04 2.58) and there was a trend toward higher odds of externalizing behavior (AOR: 1.56; 95% CI: 0.98-2.49). CONCLUSIONS: Overall, 20% of the mothers in our sample of at-risk families had a RRB, which was far greater than the national average of 11%. RRB was associated with a greater likelihood of adverse consequences for both the mother and the index child. The lack of program effects can be traced to shortcomings in the program's design and implementation system. HSP contracts required only that family planning be introduced any time during a family's first year of enrollment. Because conception can occur very soon after the index birth, a better design would be to introduce family planning counseling early in a family's enrollment in home visiting. Another shortcoming was that, although fathers could be included in counseling, they took part in only approximately one fifth of home visits. It is possible that program effects were attenuated in families in which the father wanted a child. In conclusion, the Hawaii HSP did not reduce RRB or alter its malleable determinants. RRB was associated with adverse outcomes for both the mother and the index child. This is particularly relevant for this population of families that are already at risk for child maltreatment, for which we have found parenting stress to be associated with abusive parenting behavior by the mother. Our findings support and broaden the rationale for the Healthy People 2010 objective to reduce RRB. We think our findings are valuable for guiding the future development of home visiting in general and this widely replicated paraprofessional model in particular. PMID- 15342893 TI - Low-normal gestational age as a predictor of asthma at 6 years of age. AB - BACKGROUND: Perinatal factors, including gestational age and birth weight, influence the development of atopy in early life. However, the role of these factors in the development of asthma in later life among children who do not develop perinatal respiratory disease remains unclear. METHODS: Four hundred fifty-four infants who had a history of allergy or asthma in at least 1 parent, were born in the 36th week of gestation or later, and did not develop perinatal respiratory distress were monitored for at least 6 years. Associations between predictor variables and asthma and wheeze were assessed with multivariate logistic regression and repeated-event analyses. RESULTS: Although we previously observed a relationship between low birth weight and persistent wheeze in the first 1 year of life, we did not observe similar associations between low birth weight and asthma at 6 years of age (odds ratio [OR]: 1.05; 95% confidence interval [CI]: 0.40-2.73). However, a strong relationship was found between low normal gestational age and asthma at 6 years of age (OR: 4.7; 95% CI: 2.1-10.5). The effects of low-normal gestational age were significantly greater among boys than among girls (boys: OR: 8.15; 95% CI: 2.98-22.3; girls: OR: 1.90; 95% CI: 0.38-13.83). Longitudinal analysis of the relationship between gestational age and wheeze during the 6 years of observation confirmed these gender differences. CONCLUSIONS: Among children at high risk of developing atopic disease, late prematurity might be an important additional determinant of asthma later in life, and these effects are gender specific. PMID- 15342894 TI - Computer-aided anthropometry in the evaluation of dysmorphic children. AB - OBJECTIVE: Although anthropometry is widely regarded as the technique of choice for the evaluation of dysmorphic features among children, it is only rarely used in clinical practice. Possible reasons for this are the limited access of pediatricians to appropriate reference values and the time-consuming way measurements must be plotted onto growth charts. This article describes a computer program that overcomes these problems and greatly facilitates the use of anthropometric measurements in pediatric medicine. DESIGN: A computer program for PalmOS-based, handheld computers that compares entered anthropometric measurements with age- and gender-matched reference values was developed. The software is distributed free of charge and can be downloaded (www.medgen.unizh.ch/abase/). RESULTS: The ABase software contains age- and gender-matched reference values for 18 anthropometric measures frequently used for the evaluation of dysmorphic children. The proband's age is automatically calculated from the entered birth date and can be corrected for prematurity. Entered measurements are compared with the internal reference values; the results are displayed as percentile ranks or z scores and are plotted on digital growth charts. CONCLUSIONS: The presented software makes anthropometric reference values easily accessible on the ward or in the clinic and greatly reduces the time needed to plot the measurements onto growth charts. This should promote more widespread use of anthropometry in pediatric medicine. PMID- 15342895 TI - Prediction models for evaluation of total-body bone mass with dual-energy X-ray absorptiometry among children and adolescents. AB - OBJECTIVE: The performance of dual-energy x-ray absorptiometry (DXA) in identifying children with decreased bone mass is increasing, but there is no consensus regarding how to interpret the results. The World Health Organization diagnostic categories for normal, osteopenia, and osteoporosis, based on T scores, are not applicable to children and adolescents who have not yet reached peak bone mass. The pediatric reference standards provided by DXA manufacturers have been questioned. Bone mineral density determined with DXA is "areal" density (a 2-dimensional measurement of a 3-dimensional structure), and its misleading nature among growing and maturing children is well recognized. Few published pediatric reference values for bone mineral density measured with DXA include factors that are known to affect the results besides age and gender. Our objective was to develop an algorithm for the evaluation of bone mass among children that included known determinants of bone mass and of its measurement with DXA. METHODS: Height, weight, pubertal status, and total-body bone mineral content, total-body bone area, and total-body bone mineral density measured with DXA were recorded for an ethnically diverse group of healthy pediatric subjects (n = 1218; age: 6-18 years). Prediction models for bone measurements were developed and validated with healthy pediatric subjects and then applied to children with medical disorders. RESULTS: There was a significant gender effect, as well as an interaction between gender and ethnicity. Separate models were developed for log total-body bone mineral content, log total-body bone area, and 1/total-body bone mineral density for girls and boys. The variability explained for each measurement increased from level 1, including age and ethnicity (76 86%), to level 2, including age, ethnicity, height, and weight (84-97%), and to level 3, including age, ethnicity, height, weight, and bone area (89-99%). Pubertal stage was an additional significant predictor of bone measurements but increased the explained variability by only 0.1% with height and weight in the models. The values predicted with each model were not different from measured values for the validation group but were different for patients with medical disorders, with different patterns according to the diagnoses. CONCLUSIONS: These models, including known determinants of bone mass and of bone measurements with DXA, provide an evaluation of pediatric bone mass that proceeds in steps from level 1 to level 3. The outcomes were different for patients at risk for compromised bone mass, compared with healthy children, with specific patterns for each medical disorder. We propose an algorithm for evaluation of bone measurements that follows levels 1 to 3. Our findings suggest that application of this algorithm to well-characterized groups of pediatric patients could identify disease-specific features of DXA results. We recommend this approach as a basis for consensus regarding the clinical evaluation of pediatric bone mass, and we suggest that it could lead to meaningful classification of pediatric bone disorders, investigation of pathophysiologic processes, and development of appropriate interventions. PMID- 15342896 TI - Nonadherence with pediatric human immunodeficiency virus therapy as medical neglect. AB - OBJECTIVE: To examine the results of an interventionist approach applied to human immunodeficiency virus (HIV)-infected children for whom caregiver nonadherence was suspected as the cause of treatment failure. METHODS: The medical records of a cohort of 16 perinatally HIV-infected children whose care was managed at the Arkansas Children's Hospital Pediatric HIV Clinic for an uninterrupted period of >or=3 years were reviewed through July 2003. Data collected included date of birth, dates of and explanations for clinic visits and hospitalizations, dates of laboratory evaluations, CD4(+) T cell percentages, plasma HIV-1 RNA levels, antiretroviral medications, viral resistance tests (eg, phenotype and genotype), and physician-initiated interventions to enhance adherence to the medication regimen. A stepwise interventionist approach was undertaken when patients continued to demonstrate high viral loads, despite documented viral sensitivity to the medication regimen and caregivers' insistence that medications were being administered regularly. Step 1 was prescribing a home health nurse referral, step 2 was administering directly observed therapy (DOT) while the patient was hospitalized for 4 days, and step 3 was submitting a physician-initiated medical neglect report to the Arkansas Department of Human Services. RESULTS: The results for 6 patients for whom this stepwise approach was initiated are reported. Home health nurse referrals failed to result in sustained improvements in adherence in all 6 cases. Viral load assays performed before and after DOT provided an objective measure of the effect of adherence, with 12 hospitalizations resulting in a mean +/- SD decrease in HIV RNA levels of 1.09 +/- 0.5 log(10) copies per mL, with a range of 0.6 to 2.1 log(10) copies per mL. Four families responded to DOT hospitalization, and sustained decreases in the respective patients' viral loads were noted. In 2 cases, medical neglect reports were submitted when DOT did not result in improved adherence. These patients were eventually placed in foster care, with subsequent improvements in their viral loads and CD4(+) T cell percentages. CONCLUSIONS: Nonadherence with antiretroviral therapy can be established on the basis of persistently elevated HIV RNA levels that decrease with DOT. Nonadherence poses a danger to the child that is grave and potentially irreversible. Caregivers should be offered all available resources to help them adhere to a sound treatment plan. In cases of demonstrated inability to provide needed care, it is necessary to consider seeking child protection, even for apparently healthy children. PMID- 15342897 TI - Placement changes and emergency department visits in the first year of foster care. AB - OBJECTIVES: Despite great needs, many children in foster care do not receive adequate medical care. Suboptimal care may be attributable in part to placement changes, which disrupt continuity of care by both a consistent surrogate parent and potentially a primary care physician. These disruptions in turn may lead to increased use of the emergency department (ED) for outpatient care. The primary aim of this study was to test whether a greater rate of placement changes was associated with increasing use of the ED among children in their first year of foster care. The secondary aim was to compare ED visit rates with rates of visits to other ambulatory care settings among children in foster care and other Medicaid-eligible children not in foster care. METHODS: Using Medicaid claims linked to foster care administrative data, we assembled a retrospective cohort of foster children in a large urban municipality from 1993 to 1996. Eligible children spent at least 9 months in a new episode of foster care and were continuously eligible for Medicaid during a 1-year follow-up period. A comparison cohort was drawn from Medicaid-eligible children not in foster care during fiscal year 1995. The dependent variable was the rate of visits to the ED or other ambulatory care settings during a 1-year follow-up period. A negative binomial model estimated visit rates to the ED and other ambulatory care settings as the number of foster care placements increased. Potential interactions were considered between age and location of service use (ED or ambulatory care setting), between age and foster care placements, and between location of service use and number of foster care placements. RESULTS: The 2358 children in the sample accounted for 1206 ED visits during the follow-up period; 38% experienced >or=2 placement changes. Children of all ages exhibited increasing reliance on the ED for ambulatory care services as the number of placements increased, with the rates of ED use more than doubling for all age groups beyond infancy. However, other ambulatory care service use increased by only 41% to 53%; there was much less utilization in these nonemergency settings for all children but particularly toddlers and infants, compared with their Medicaid-eligible peers. Although the absolute rates of all visits for younger foster children (<6 years of age) were low, older foster children had increasingly greater ED use than did their Medicaid-eligible peers, with nearly double the rate of ED visits in the adolescent age group. A temporal relationship between placements and ED visits was also identified; 75% of ED visits occurring within 3 weeks of a placement change occurred in the period after a placement change. CONCLUSIONS: Foster children received fewer overall outpatient services than did their Medicaid eligible peers, but with age and increasing numbers of placements, had higher visit rates and received a greater proportion of their overall outpatient care in the ED. These results suggest that poor access to nonemergent ambulatory care settings might have contributed to an increasing reliance on ED settings as foster care placements increased. The temporal relationship between ED visits and placement changes underscores the need for better health care management for foster children, particularly in the period after placement changes. PMID- 15342898 TI - Circumstances surrounding the deaths of hospitalized children: opportunities for pediatric palliative care. AB - OBJECTIVES: Little is known regarding the assessment and treatment of symptoms during end-of-life (EOL) care for children. This study was conducted to describe the circumstances surrounding the deaths of hospitalized terminally ill children, especially pain and symptom management by the multidisciplinary pediatric care team. DESIGN: Patients in the neonatal intensive care unit, pediatric critical care unit, or general pediatric units of Vanderbilt Children's Hospital who were hospitalized at the time of death, between July 1, 2000, and June 30, 2001, were identified. Children eligible for the survey had received inpatient EOL care at the hospital for at least 24 hours before death. A retrospective medical record review was completed to describe documentation of care for these children and their families during the last 72 hours of life. RESULTS: Records of children who had received inpatient EOL care were identified (n = 105). A majority (87%) of children were in an intensive care setting at the time of death. Most deaths occurred in the pediatric critical care unit (56%), followed by the neonatal intensive care unit (31%). Pain medication was received by 90% of the children in the last 72 hours of life, and 55% received additional comfort care measures. The presence of symptoms other than pain was infrequently documented. CONCLUSIONS: The duration of hospitalization for most children dying in this inpatient setting was sufficient for provision of interdisciplinary pediatric palliative care. Management of pain and other symptoms was accomplished for many children. The documentation of pain and symptom assessment and management can be improved but requires new tools. PMID- 15342899 TI - Uncomplicated intraventricular hemorrhage is followed by reduced cortical volume at near-term age. AB - BACKGROUND: Intraventricular hemorrhage (IVH) is the most common brain injury among premature infants. Neonates with IVH are at greater risk of impaired neurodevelopmental outcomes, compared with those without IVH. IVH causes destruction of the germinal matrix and glial precursor cells, with possible effects on cortical development. OBJECTIVE: To investigate cortical development after uncomplicated IVH (with no parenchymal involvement and no posthemorrhagic hydrocephalus). We hypothesized that uncomplicated IVH would be followed by reduced cortical volume among premature infants at near-term age. METHODS: A prospective cohort study was conducted, with preset selection criteria. Infants with small-for-gestational age birth weight, congenital abnormalities or brain malformations, metabolic disorders, recurrent sepsis, or necrotizing enterocolitis were excluded. Also, infants with posthemorrhagic hydrocephalus, parenchymal involvement of hemorrhage, cystic periventricular leukomalacia, or persistent ventriculomegaly were excluded, on the basis of routine serial ultrasonographic assessments. Three-dimensional images were acquired for 23 infants at near-term age, with 3-T magnetic resonance imaging and a magnetization prepared rapid gradient echo sequence. Image analysis and segmentation of the cerebrum in different tissue types were based on signal contrast and anatomic localization. The cortical gray matter (CGM), subcortical gray matter, white matter, and intraventricular cerebrospinal fluid volumes of 12 infants with uncomplicated IVH were compared with those of 11 infants without IVH, using multivariate analysis of variance. RESULTS: The multivariate analysis of variance for the regional brain volumes in the 2 groups indicated significance (Wilks' lambda = 0.546). The CGM volume was significantly reduced in the IVH group (no IVH group: 122 +/- 12.9 mL; IVH group: 102 +/- 14.6 mL; F = 13.218). This finding remained significant after testing for possible confounding factors and adjustment for size differences between the infants (F = 9.415). There was no difference in the volumes of subcortical gray matter, white matter, and cerebrospinal fluid. CONCLUSIONS: This is the first study to document impaired cortical development after uncomplicated IVH. The impairment was demonstrated by a 16% reduction in cerebral CGM volume at near-term age. The finding supports concerns regarding possible glial precursor cell loss after germinal matrix IVH, but its clinical significance is still unclear. The alteration in brain development demonstrated in this report supports closer neurodevelopmental follow up monitoring of preterm infants with uncomplicated IVH. PMID- 15342900 TI - Blood pressure at 6 years of age after prenatal exposure to betamethasone: follow up results of a randomized, controlled trial. AB - OBJECTIVE: To determine whether prenatal exposure to betamethasone for the prevention of neonatal respiratory distress syndrome (RDS) alters blood pressure in childhood. DESIGN: Prospective follow-up study of a randomized, double-blind, placebo-controlled trial. SETTING: National Women's Hospital (Auckland, New Zealand). PARTICIPANTS: Two hundred twenty-three 6-year-old children of mothers who presented with unplanned premature labor and took part in a randomized, controlled trial of prenatal betamethasone therapy for the prevention of neonatal RDS. INTERVENTION: Mothers received 2 doses of betamethasone (12 mg) or placebo, administered through intramuscular injection, 24 hours apart. MAIN OUTCOME MEASURES: Systolic and diastolic blood pressure at 6 years of age. RESULTS: Children exposed prenatally to betamethasone (n = 121) did not differ in systolic or diastolic blood pressure from children exposed to placebo (n = 102) (mean difference: systolic: -1.6 mm Hg; 95% confidence interval: -4.1 to 0.8 mm Hg; diastolic: -0.3 mm Hg; 95% confidence interval: -2.5 to 1.8 mm Hg). CONCLUSION: Prenatal exposure to betamethasone for prevention of neonatal RDS does not alter blood pressure at 6 years of age. PMID- 15342901 TI - "Social medication" and the control of children: a qualitative study of over-the counter medication among Australian children. AB - OBJECTIVE: The aim of the study was to identify the patterns of use of over-the counter (OTC) medications among children. METHODS: The study used a qualitative design, with in-depth interviews of 40 parents with children <5 years of age. RESULTS: There were 3 striking and readily apparent themes in the use of OTC medications among children. One was the administration of OTC medications as a form of "social medication," to give parents control over children's behavior that they perceived as fractious and irritating. A related theme was the use of OTC medications to reduce the inconvenience to the parents of having a sick child, again giving parents greater control and better time-management abilities. Finally, acetaminophen was considered by many parents to have almost miraculous properties in calming, sedating, and lifting the mood of children. CONCLUSIONS: The use of OTC medications for the treatment of minor ailments among children is widespread, despite the lack of evidence of efficacy of the most commonly used medications and the potential for toxicity. With the increasing propensity to look to medication as a means of supporting changing lifestyles, there is an urgent need to review the prevalence and effects of social medication. PMID- 15342902 TI - Temporary removal of navel piercing jewelry for surgery and imaging studies. AB - BACKGROUND: Navel piercings are becoming increasingly popular among adolescents. Body piercing jewelry must be removed before surgery, because electrical burns can occur with electrocauterization. Also, piercing jewelry creates artifacts during magnetic resonance imaging. Even temporary piercing jewelry removal may lead to closure of the subcutaneous tract; therefore, temporary replacement with a nonmetallic spacer is indicated. OBJECTIVE: This report describes a simple, safe, effective method for perioperative temporary replacement of navel piercing jewelry with a universally available, nonmetallic, sterile, intravenous catheter. METHODS: The umbilical piercing jewelry of 5 adolescent female patients was removed before surgical or radiologic interventions by using the following technique: 1) disinfecting the piercing and umbilical area with 70% isopropyl alcohol, 2) opening the piercing jewelry by removing the bead from the bar, 3) placing the tip of a tight-fitting intravenous catheter (14 or 16 gauge, without the needle) over the threaded tip of the bar, 4) advancing the intravenous catheter caudally, pushing the piercing out of the skin tract, and 5) removing the jewelry and leaving the intravenous catheter in the subcutaneous skin tract as a spacer. After the intervention, the piercing jewelry was reinserted by following the described steps in reverse order. The patients were evaluated after 1 week, for assessment of any adverse effects, including infection. RESULTS: With this method, piercing jewelry removal and reinsertion required <2 minutes for all patients. No procedural complications were noted. All jewelry was reinserted successfully after the procedure. The intravenous catheter left in situ did not interfere with surgery, and it did not produce any artifacts or adverse effects during magnetic resonance imaging. Follow-up evaluations at 1 week demonstrated no signs of irritation, infection, or other side effects of the procedure for any of the patients. CONCLUSIONS: Periprocedural removal of navel piercing jewelry with this new technique proved to be quick, feasible, and safe. Piercings in other locations on the body may be replaced for surgery or imaging studies in the same way, with minor modifications. PMID- 15342903 TI - Congenital aplastic anemia caused by mutations in the SBDS gene: a rare presentation of Shwachman-Diamond syndrome. AB - CLINICAL FINDINGS: Aplastic anemia was diagnosed at birth for a first child from healthy nonconsanguineous parents. The girl had hypoglycemia, which normalized within 2 months. Cow milk allergy was suspected initially, because of skin lesions and diarrhea, followed by severe growth retardation. Clinical and radiologic symptoms gradually became typical for Shwachman-Diamond syndrome. Two common mutations in the SBDS gene (183-184TA-->CT [K62X] and IVS2(258)+2T--> C [C84fs]) were found. RESULTS: Bone marrow transplantation from a matched unrelated donor was unsuccessful. The genetic information from the deceased patient enabled us to perform prenatal molecular studies during the subsequent pregnancy, successfully predicting a nonaffected child. CONCLUSIONS: This report describes for the first time the hematologic abnormalities of congenital aplastic anemia and prolonged neonatal hypoglycemia as the presenting symptoms of Shwachman-Diamond syndrome. The finding of common mutations in the presence of these symptoms at birth suggests the lack of a clear phenotype-genotype relationship in this syndrome. PMID- 15342904 TI - A renaissance of metabolite sensing and signaling: from modular domains to riboswitches. PMID- 15342905 TI - A network of interacting factors triggering different cell fates. PMID- 15342906 TI - Proapoptotic N-truncated BCL-xL protein activates endogenous mitochondrial channels in living synaptic terminals. AB - Neuronal death is often preceded by functional alterations at nerve terminals. Anti- and proapoptotic BCL-2 family proteins not only regulate the neuronal death pathway but also affect excitability of healthy neurons. We found that exposure of squid stellate ganglia to hypoxia, a death stimulus for neurons, causes a cysteine protease-dependent loss of full-length antiapoptotic BCL-xL, similar to previous findings in mammalian cells. Therefore, to determine the direct effect of the naturally occurring proapoptotic cleavage product of BCL-xL on mitochondria, recombinant N-truncated BCL-xL was applied to mitochondria inside the squid presynaptic terminal and to purified mitochondria isolated from yeast. N-truncated BCL-xL rapidly induced large multi-conductance channels with a maximal conductance significantly larger than those produced by full-length BCL xL. This activity required the hydrophobic C terminus and the BH3 domain of BCL xL. Moreover, N-truncated BCL-xL failed to produce any channel activity when applied to plasma membranes, suggesting that a component of the mitochondrial membrane is necessary for its actions. Consistent with this idea, the large channels induced by N-truncated BCL-xL are inhibited by NADH and require the presence of VDAC, a voltage-dependent anion channel present in the outer mitochondrial membrane. These observations suggest that the mitochondrial channels specific to full-length and N-truncated BCL-xL contribute to their opposite effects on synaptic transmission, and are consistent with their opposite effects on the cell death pathway. PMID- 15342907 TI - CD1d-restricted T cell activation by nonlipidic small molecules. AB - In addition to NK T cells expressing invariant Valpha14 or Valpha24 T cell receptors (TCRs), the CD1d-restricted T cell repertoire is comprised of T cells with diverse TCRs that mediate inflammation during autoimmune and infectious disease. Here we describe the isolation of human Valpha24(-) T cells that are activated by antigen and CD1d. Mass spectrometric and NMR studies revealed that the stimulatory compounds were neither peptidic nor lipidic but instead were composed of sulfur and aromatic hydrocarbon rings, corresponding to the general structure of phenyl pentamethyldihydrobenzofuran sulfonates. Studies of the molecular mechanism of T cell activation showed that a clonotypic Valpha2/Vbeta21 TCR transmitted activating signals, which were highly specific for hydroxylation and methylation patterns at the terminal structures of stimulatory compounds. These studies provide evidence for noninvariant CD1d-restricted T cells in humans and identify the complete molecular structure of a nonlipidic small molecule that activates T cells through an alphabeta TCR. PMID- 15342908 TI - Mast cells: a unique source of renin. AB - In addition to the traditional renin-angiotensin system, a great deal of evidence favors the existence of numerous independent tissue-specific renin-angiotensin systems. We report that mast cells are an additional source of renin and constitute a unique extrarenal renin-angiotensin system. We use renin-specific antibodies to demonstrate that cardiac mast cells contain renin. Extending this observation to the human mast cell line HMC-1, we show that these mast cells also express renin. The HMC-1 renin RT-PCR product is 100% homologous to Homo sapiens renin. HMC-1 cells also contain renin protein, as demonstrated both by immunoblot and immunocytochemical analyses. Renin released from HMC-1 cells is active; furthermore, HMC-1 cells are able to synthesize renin. It is known that, in the heart, mast cells are found in the interstitium in close proximity to nerves and myocytes, which both express angiotensin II receptors. Inasmuch as myocardial interstitium contains angiotensinogen and angiotensin-converting enzyme, and because we were able to detect renin only in mast cells, we postulate that the release of renin from cardiac mast cells is the pivotal event triggering local formation of angiotensin II. Because of the ubiquity of mast cells, our results represent a unique paradigm for understanding local renin-angiotensin systems, not just in the heart, but in all tissues. Our findings provide a rationale for targeting mast cells in conjunction with renin-angiotensin system inhibitors in the management of angiotensin II-related dysfunctions. PMID- 15342909 TI - Chromosome painting using repetitive DNA sequences as probes for somatic chromosome identification in maize. AB - Study of the maize (Zea mays L.) somatic chromosomes (2n = 20) has been difficult because of a lack of distinguishing characteristics. To identify all maize chromosomes, a multicolor fluorescence in situ hybridization procedure was developed. The procedure uses tandemly repeated DNA sequences to generate a distinctive banding pattern for each of the 10 chromosomes. Fluorescence in situ hybridization screening trials of nonsubtracted or subtracted PCR libraries resulted in the isolation of microsatellite 1-26-2, subtelomeric 4-12-1, and 5S rRNA 2-3-3 clones. These three probes, plus centromeric satellite 4 (Cent4), centromeric satellite C (CentC), knob, nucleolus-organizing region (NOR), pMTY9ER telomere-associated sequence, and tandemly repeated DNA sequence 1 (TR-1) were used as a mixture for hybridization to root-tip chromosomes. All 10 chromosomes were identified by the banding and color patterns in the 14 examined lines. There was significant quantitative variation among lines for the knob, microsatellite, TR-1, and CentC signals. The same probe mixture identifies meiotic pachytene, late prophase I, and metaphase I chromosomes. The procedure could facilitate the study of chromosomal structure and behavior and be adapted for other plant species. PMID- 15342910 TI - Deletion of Bax eliminates sex differences in the mouse forebrain. AB - Several of the best-studied sex differences in the mammalian brain are ascribed to the hormonal control of cell death. This conclusion is based primarily on correlations between pyknotic cell counts in development and counts of mature neurons in adulthood; the molecular mechanisms of hormone-regulated, sexually dimorphic cell death are unknown. We asked whether Bax, a member of the Bcl-2 family of proteins that is required for cell death in many developing neurons, might be essential for sex differences in neuron number. We compared Bax knockout mice and their WT siblings, focusing on two regions of the mouse forebrain that show opposite patterns of sexual differentiation: the principal nucleus of the bed nucleus of the stria terminalis, in which males have more neurons than do females, and the anteroventral periventricular nucleus (AVPV), where females have more neurons overall and many more dopaminergic neurons than do males. Testosterone, or its metabolites, is responsible for the sex differences in both nuclei. A null mutation of the Bax gene completely eliminated sex differences in overall cell number in both the principal nucleus of the bed nucleus of the stria terminalis and AVPV. Thus, Bax-dependent cell death is required for sexual differentiation of cell number, regardless of whether testosterone decreases or increases cell death. In contrast, the sex difference in AVPV dopaminergic cell number, as measured by tyrosine hydroxylase immunohistochemistry, was not affected by Bax gene deletion, demonstrating heterogeneity of mechanisms controlling cell number within a single nucleus. PMID- 15342911 TI - Naturally occurring H-DNA-forming sequences are mutagenic in mammalian cells. AB - Naturally occurring DNA sequences can form noncanonical structures such as H-DNA, which are abundant and regulate the expression of several disease-linked genes. Here, we show that H-DNA-forming sequences are intrinsically mutagenic in mammalian cells. This finding suggests that DNA is a causative factor in mutagenesis and not just the end product. By using the endogenous H-DNA-forming sequence found in the human c-myc promoter, mutation frequencies in a reporter gene were increased approximately 20-fold over background in COS-7 cells. H-DNA induced double-strand breaks (DSBs) were detected near the H-DNA locus. The structures of the mutants revealed microhomologies at the breakpoints, consistent with a nonhomologous end-joining repair of the DSBs. These results implicate H DNA-induced DSBs in c-myc gene translocations in diseases such as Burkitt's lymphoma and t(12;15) BALB/c plasmacytomas, where most breakpoints are found near the H-DNA-forming site. Thus, our findings suggest that H-DNA is a source of genetic instability resulting from DSBs and demonstrate that naturally occurring DNA sequences are mutagenic in mammals, perhaps contributing to genetic evolution and disease. PMID- 15342912 TI - Proteasomal degradation of the FoxO1 transcriptional regulator in cells transformed by the P3k and Akt oncoproteins. AB - The P3k oncoprotein [homolog of the catalytic subunit p110alpha of class 1A phosphoinositide 3-kinase (PI3K)] and its downstream effector Akt induce oncogenic transformation in cultures of chicken embryo fibroblasts (CEF). The winged helix transcription factor FoxO1 is a growth-attenuating and proapoptotic protein and serves as a substrate of Akt. Here we show that FoxO1 expression is constitutively suppressed in CEF transformed by P3k or Akt. The FoxO1 protein level is high in serum-starved normal CEF, but platelet-derived growth factor treatment induces rapid phosphorylation and disappearance of FoxO1. PI3K inhibitors or the proteasome inhibitor lactacystin interfere with this process. These data suggest that phosphorylation-dependent degradation of FoxO1 by means of proteasomes plays a role in oncogenic transformation by P3k and Akt. A dominant negative mutant of FoxO1 containing the repressor domain of the Drosophila Engrailed protein induces partial oncogenic transformation of CEF and interferes with FoxO1-dependent transcriptional activation. The FoxG1 oncoprotein also inhibits transcriptional activation by FoxO1. Inhibition of FoxO1, albeit by different mechanisms, appears to be a common denominator of the PI3K and FoxG1 oncogenic pathways. PMID- 15342913 TI - A cationic lipid-formulated plasmid DNA vaccine confers sustained antibody mediated protection against aerosolized anthrax spores. AB - DNA vaccines provide an attractive technology platform against bioterrorism agents due to their safety record in humans and ease of construction, testing, and manufacture. We have designed monovalent and bivalent anthrax plasmid DNA (pDNA) vaccines encoding genetically detoxified protective antigen (PA) and lethal factor (LF) proteins and tested their immunogenicity and ability to protect rabbits from an aerosolized inhalation spore challenge. Immune responses after two or three injections of cationic lipid-formulated PA, PA plus LF, or LF pDNAs were at least equivalent to two doses of anthrax vaccine adsorbed (AVA). High titers of anti-PA, anti-LF, and neutralizing antibody to lethal toxin (Letx) were achieved in all rabbits. Eight or nine animals in each group were challenged with 100x LD(50) of aerosolized anthrax spores 5 or 9 weeks after vaccination. An additional 10 animals vaccinated with PA pDNA were challenged >7 months postvaccination. All animals receiving PA or PA plus LF pDNA vaccines were protected. In addition, 5 of 9 animals receiving LF pDNA survived, and the time to death was significantly delayed in the others. Groups receiving three immunizations with PA or PA plus LF pDNA showed no increase in anti-PA, anti-LF, or Letx neutralizing antibody titers postchallenge, suggesting little or no spore germination. In contrast, titer increases were seen in AVA animals, and in surviving animals vaccinated with LF pDNA alone. Preclinical evaluation of this cationic lipid-formulated bivalent PA and LF vaccine is complete, and the vaccine has received U.S. Food and Drug Administration Investigational New Drug allowance. PMID- 15342914 TI - DNA heterogeneity and phosphorylation unveiled by single-molecule electrophoresis. AB - Broad-spectrum analysis of DNA and RNA samples is of increasing importance in the growing field of biotechnology. We show that nanopore measurements may be used to assess the purity, phosphorylation state, and chemical integrity of nucleic acid preparations. In contrast with gel electrophoresis and mass spectrometry, an unprecedented dynamic range of DNA sizes and concentrations can be evaluated in a single data acquisition process that spans minutes. Because the molecule information is quantized and digitally recorded with single-molecule resolution, the sensitivity of the system can be adjusted in real time to detect trace amounts of a particular DNA species. PMID- 15342915 TI - Cell-type-specific binding of the transcription factor CREB to the cAMP-response element. AB - The cAMP-response element-binding protein (CREB) transcription factor was initially identified as a mediator of cAMP-induced gene expression. CREB binds to a target sequence termed the cAMP-response element (CRE) found in many cellular and viral gene promoters. One of the best-characterized CREs resides in the promoter of the gene encoding the neuropeptide somatostatin, and this element has served as a model for studies of CREB function. Phosphorylation of CREB by protein kinase A allows recruitment of the coactivator CREB-binding protein (CBP). A central tenet of the CREB-CBP model is that CREB binds constitutively to the CRE and that regulation occurs through the phosphorylation-dependent recruitment of CBP. In this report, we use chromatin immunoprecipitation assays to show that CREB does not interact in vivo with the somatostatin CRE, or similar elements in several other genes, in PC12 cells, a standard model for studies of CREB function. Rather, CREB binding in vivo is regulated in a cell-specific manner, a finding that was confirmed by using in vivo genomic footprinting assays. The CREs in other genes were also found to interact differentially with CREB in PC12 cells, hepatoma cells, and cortical neurons. We conclude that the family of CREB target genes differs from one cell type to another and that the ability of CREB to bind to a particular CRE represents an important component of gene regulation. PMID- 15342916 TI - Delayed neuropathology after carbon monoxide poisoning is immune-mediated. AB - The neuropathological sequelae of carbon monoxide (CO) poisoning cannot be explained by hypoxic stress alone. CO poisoning also causes adduct formation between myelin basic protein (MBP) and malonylaldehyde, a reactive product of lipid peroxidation, resulting in an immunological cascade. MBP loses its normal cationic characteristics, and antibody recognition of MBP is altered. Immunohistochemical evidence of degraded MBP occurs in brain over days, along with influx of macrophages and CD-4 lymphocytes. Lymphocytes from CO-poisoned rats subsequently exhibit an auto-reactive proliferative response to MBP, and there is a significant increase in the number of activated microglia in brain. Rats rendered immunologically tolerant to MBP before CO poisoning exhibit acute biochemical changes in MBP but no lymphocyte proliferative response or brain microglial activation. CO poisoning causes a decrement in learning that is not observed in immunologically tolerant rats. These results demonstrate that delayed CO-mediated neuropathology is linked to an adaptive immunological response to chemically modified MBP. PMID- 15342917 TI - Tumor-promoting phorbol esters and activated Ras inactivate the tuberous sclerosis tumor suppressor complex via p90 ribosomal S6 kinase. AB - Tuberous sclerosis complex (TSC) is a genetic disorder caused by mutations in either of the two tumor suppressor genes TSC1 or TSC2, which encode hamartin and tuberin, respectively. Tuberin and hamartin form a complex that inhibits signaling by the mammalian target of rapamycin (mTOR), a critical nutrient sensor and regulator of cell growth and proliferation. Phosphatidylinositol 3-kinase (PI3K) inactivates the tumor suppressor complex and enhances mTOR signaling by means of phosphorylation of tuberin by Akt. Importantly, cellular transformation mediated by phorbol esters and Ras isoforms that poorly activate PI3K promote tumorigenesis in the absence of Akt activation. In this study, we show that phorbol esters and activated Ras also induce the phosphorylation of tuberin and collaborates with the nutrient-sensing pathway to regulate mTOR effectors, such as p70 ribosomal S6 kinase 1 (S6K1). The mitogen-activated protein kinase (MAPK) activated kinase, p90 ribosomal S6 kinase (RSK) 1, was found to interact with and phosphorylate tuberin at a regulatory site, Ser-1798, located at the evolutionarily conserved C terminus of tuberin. RSK1 phosphorylation of Ser-1798 inhibits the tumor suppressor function of the tuberin/hamartin complex, resulting in increased mTOR signaling to S6K1. Together, our data unveil a regulatory mechanism by which the Ras/MAPK and PI3K pathways converge on the tumor suppressor tuberin to inhibit its function. PMID- 15342918 TI - Osteoarthritis. AB - In this case study, we describe the symptoms, evaluation, and management of a woman with osteoarthritis (OA). OA is the most common form of arthritis worldwide and it is a major cause of disability in the elderly. Although there are several aging-related changes in the musculoskeletal system that may contribute to the pathogenesis of this disease, research suggests that OA is not merely an inevitable result of aging. OA is most likely a multifactorial process whereby non-aging-related factors also contribute to the onset, progression, and symptomatology of the disease. Specifically, both biochemical factors, including physiological properties of cartilage and bone, and biomechanical factors such as muscle strength, proprioception, and joint loading have been implicated in the pathogenesis of OA. Newer nonpharmacological treatment options are focusing on how to improve symptoms and prevent progression of the disease through mechanical interventions. PMID- 15342919 TI - Stuck in the craw.Parkinson's proteins choke cell's recycling system. PMID- 15342920 TI - Hastening death to delay aging. New approach speeds hunt for manipulations that retard fly decrepitude . PMID- 15342921 TI - Regenerating regeneration.Salamanders, flatworms, and other creatures engage in dazzling feats of renewal. Now, researchers are beginning to uncover the molecular bases for these body-building tricks, hoping to decipher how humans might perform similar stunts. PMID- 15342922 TI - Deja vu. The history of in vitro fertilization could offer insights into how the controversy over the use of human embryos for stem cell research might play out. PMID- 15342923 TI - Not wisely but too well: aging as a cost of neuroendocrine activity. AB - Progressive decline of some neuroendocrine signaling systems has long been assumed to cause age-related physiological impairments and limit life span. However, hypophysectomy--removal of the pituitary gland--can delay many aspects of the aging process, and recent genetic studies have confirmed that reducing the secretion of pituitary hormones can increase the life span of laboratory organisms. Most strikingly, reducing activity of the insulin/insulin-like growth factor-1 signaling system substantially increases life span. Conversely, activity of the reproductive system or activation of stress responses can curtail life span. Because caloric restriction also reduces the activity of several neuroendocrine systems while increasing life span, it now appears that the aging process is driven, at least in part, by neuroendocrine activity rather than by its decline with age. PMID- 15342924 TI - Aging of the human adrenal cortex. AB - The most striking age-related change in the human adrenal cortex is the decline in secretion of dehydroepiandrosterone and its sulfate, steroids synthesized by the inner zone of the cortex, the zona reticularis. Because these steroids are of essentially unknown function, the importance of this age-related change is the subject of considerable debate. It is likely that the age-related change in these steroids results from loss of zona reticularis cells or impairment of their function. During aging, cumulative damage to the zona reticularis could occur through ischemia-related infarcts and other causes of cell death. Cellular senescence could contribute to a loss of the ability of the tissue to replace lost cells. In contrast, feedback mechanisms that regulate adrenocortical growth cause compensatory local tissue hyperplasias called nodules. The effect of imperfect repair of damage combined with compensatory overgrowth in the form of nodules leads to an increasingly abnormal tissue architecture. PMID- 15342925 TI - The relationship between disease activity and quality of life in systemic lupus erythematosus. AB - OBJECTIVE: To determine the quality of life (QOL) in SLE patients and correlate it with disease activity. METHODS: Lupus patients fulfilling the ACR 1997 criteria for SLE were included in this cross-sectional study. Patients were administered the World Health Organization Quality of Life-Bref (WHOQOL-Bref) to assess their quality of life. Disease activity was measured using Mexican Systemic Lupus Erythematosus Disease Activity Index (Mex-SLEDAI). RESULTS: The study group comprised 73 lupus patients (70 females and three males) with mean age 35.22 +/- 11.15 yr and mean disease duration 5.62 +/- 5.14 yr. Mean Mex SLEDAI score was 3.31 +/- 3.19. Higher disease activity scores were associated with lower QOL scores in the physical (P = 0.001) and psychological domains (P = 0.01) but showed no significant correlation with the domains of social and environmental QOL. Patients with clearly active and probably active disease showed significantly lower scores in the physical (P = 0.01) and psychological (P = 0.02) domains than patients with inactive disease. However, no significant difference was found in the domains of social and environmental QOL. Age or disease duration did not affect the QOL in any of the domains. CONCLUSIONS: Physical and psychological QOL are impaired to a larger extent in active lupus. However, social and environmental QOL do not correlate with the disease activity status in lupus patients. PMID- 15342926 TI - Muscle involvement in juvenile idiopathic arthritis. AB - OBJECTIVE: An observational study of changes in muscle structure and the relation to muscle strength in juvenile idiopathic arthritis (JIA). METHODS: Fifteen children and teenagers (eight girls and seven boys) with JIA, aged 9-19 yr (mean age 16.1), were studied. Muscle biopsies were obtained from the anterior tibial muscle and were examined using histopathological and immunohistochemical methods. Muscle fibre types were classified and fibre areas measured. As markers of inflammation, the major histocompatibility complex (MHC) class I and class II and the membrane attack complex (MAC) were analysed. Results were compared with biopsies from the gastrocnemius muscle in 33 young (19-23 yr) healthy controls. Isometric and isokinetic muscle strengths were measured in ankle dorsiflexion. Strength was compared with reference values for healthy age-matched controls. Nerve conduction velocities were recorded in the peroneal and sural nerves. RESULTS: Four of the 15 muscle biopsies were morphologically normal. Eleven biopsies showed minor unspecific changes. Two of these also showed minor signs of inflammation. MHC class II expression was found in 4/15 patients, which was significantly more than in the healthy controls (P = 0.0143). The expression of MHC class I and MAC did not differ from that in the controls. The mean area of type I fibres was lower than that of type IIA fibres in 12/13 biopsies. Muscle strength was significantly reduced in the patient group. There was a significant positive correlation between muscle fibre area and muscle strength. Nerve conduction studies were normal in all cases. CONCLUSIONS: Changes in leg muscle biopsies appear to be common in children and teenagers with JIA. The presence of inflammatory cells in the muscle and expression of MHC class II on muscle fibres may be a sign of inflammatory myopathy. There are no findings of type II muscle fibre hypotrophy or neuropathy, as in adults with RA. PMID- 15342927 TI - Predictors of neuropsychiatric damage in systemic lupus erythematosus: data from the Maryland lupus cohort. AB - OBJECTIVE: To identify factors predictive of significant neuropsychiatric (NP) damage in systemic lupus erythematosus (SLE). METHODS: One hundred and thirty patients with SLE were followed at the University of Maryland Lupus Clinic from 1992 until 2003. NP manifestations were defined according to the revised American College of Rheumatology (ACR) nomenclature and case definitions for NP-SLE syndromes. Disease activity was measured using the SLE Disease Activity Index (SLEDAI), organ damage using the Systemic Lupus International Collaborating Clinics Damage Index SLICC/ACR (SDI); NP damage (NPDI) was measured with the corresponding domain of the SDI. At end of study period, 64 patients exhibited no NP damage (NPDI = 0) and 66 patients developed significant NP damage (defined as NPDI > or =1). The baseline features for these two patient groups were compared, and variables found to be significantly different were examined by multivariable analyses to determine their contribution to NP damage. RESULTS: Significant NP damage is common in SLE; mortality is infrequent and the cause of death is unrelated to NP damage. Independent predictors of significant NP damage were disease activity, Caucasian ethnicity and the presence of antiphospholipid antibodies and anti-Ro/SSA antibody. Certain clinical features at baseline predicted specific NP damage. For example, higher disease activity at baseline was predictive of psychosis and cognitive impairment, anti-dsDNA was predictive of polyneuropathy, and antiphospholipid antibodies were predictive of seizures and cerebrovascular accidents. CONCLUSIONS: In this longitudinal SLE cohort, significant cumulative NP damage occurred. Early aggressive therapy targeted towards NP manifestations may prevent the occurrence of NP damage. PMID- 15342929 TI - Human placenta-derived cells have mesenchymal stem/progenitor cell potential. AB - Mesenchymal stem/progenitor cells (MSCs) are widely distributed in a variety of tissues in the adult human body (e.g., bone marrow [BM], kidney, lung, and liver). These cells are also present in the fetal environment (e.g., blood, liver, BM, and kidney). However, MSCs are a rare population in these tissues. Here we tried to identify cells with MSC-like potency in human placenta. We isolated adherent cells from trypsin-digested term placentas and established two clones by limiting dilution. We examined these cells for morphology, surface markers, gene expression patterns, and differentiation potential and found that they expressed several stem cell markers, hematopoietic/ endothelial cell-related genes, and organ-specific genes, as determined by reverse transcription polymerase chain reaction and fluorescence-activated cell sorter analysis. They also showed osteogenic and adipogenic differentiation potentials under appropriate conditions. We suggest that placenta-derived cells have multilineage differentiation potential similar to MSCs in terms of morphology, cell-surface antigen expression, and gene expression patterns. The placenta may prove to be a useful source of MSCs. PMID- 15342930 TI - Specific knockdown of Oct4 and beta2-microglobulin expression by RNA interference in human embryonic stem cells and embryonic carcinoma cells. AB - We have used RNA interference (RNAi) to downregulate beta2-microglobulin and Oct4 in human embryonal carcinoma (hEC) cells and embryonic stem (hES) cells, demonstrating that RNAi is an effective tool for regulating specific gene activity in these human stem cells. The knockdown of Oct4 but not beta2 microglobulin expression in both EC and ES cells resulted in their differentiation, as indicated by a marked change in morphology, growth rate, and surface antigen phenotype, with respect to SSEA1, SSEA3, and TRA-1-60 expression. Expression of hCG and Gcm1 was also induced following knockdown of Oct4 expression, in both 2102Ep hEC cells and in H7 and H14 hES cells, consistent with the conclusion that, as in the mouse, Oct4 is required to maintain the undifferentiated stem cell state, and that differentiation to trophectoderm occurs in its absence. NTERA2 hEC cells also differentiated, but not to trophectoderm, suggesting their equivalence to a later stage of embryogenesis than other hEC and hES cells. PMID- 15342931 TI - Generation of tyrosine hydroxylase positive neurons from human embryonic stem cells after coculture with cellular substrates and exposure to GDNF. AB - Tyrosine hydroxylase (TH)-positive neurons were generated from human embryonic stem (hES) cells by coculturing on astrocytes or PA6 stromal cells. After 3 to 4 weeks in culture, TH-positive cells with neuronal morphology developed. Coculture with astrocytes from the embryonic striatum produced a larger number of TH positive cells than did coculture with astrocytes from embryonic mesencephalon (329 +/- 149 versus 33 +/- 16 TH-positive cells per well, p < .05). In other experiments using PA6 cells as a substrate, glial-derived neurotrophic factor (GDNF) was added to the media of differentiating hES cells, and this led to a doubling of the number of TH-positive cells (PA6: 443 +/- 105 TH-positive cells per well versus PA6 + GDNF: 934 +/- 136, p < .05). We conclude that substrates of striatal astrocytes and PA6 cells can promote differentiation of human embryonic stem cells to a TH-positive phenotype and that GDNF can increase the number of cells expressing that phenotype. PMID- 15342932 TI - Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion. AB - Human marrow stromal cells (MSCs) can be isolated from bone marrow and differentiate into multiple tissues in vitro and in vivo. These properties make them promising tools in cell and gene therapy. The lack of a specific MSC marker and the low frequency of MSCs in bone marrow necessitate their isolation by in vitro expansion prior to clinical use. This may severely reduce MSC proliferative capacity to the point that the residual proliferative potential is insufficient to maintain long-term tissue regeneration upon reinfusion. In this study we determined the effect of in vitro expansion on the replicative capacity of MSCs by correlating their rate of telomere loss during in vitro expansion with their behavior in vivo. We report that even protocols that involve minimal expansion induce a rapid aging of MSCs, with losses equivalent to about half their total replicative lifespan. PMID- 15342933 TI - Induction of tolerance in quadruple chimeric mice. AB - Human cord blood (CB) contains hematopoietic stem cells and progenitors. Because the major limitation to a widespread use of CB for transplantation lies in its limited volume, it is necessary to combine the CB from several donors. In this study, we show that lethally irradiated mice can be reconstituted with the injection of a mixture of T cell-depleted bone marrow cells (BMCs; total, 3 x 10(6)) obtained from three fully allogeneic mouse strains in two different mouse combinations. A higher survival rate was obtained in the triple injection group than in mice injected with BMCs (1 x10(6)) obtained from a single mouse strain. In the mixed chimeric mice, three kinds of donor-type and recipient-type cells were detected in all the hematopoietic organs 1 month after bone marrow transplantation (BMT). Mixed-lymphocyte reaction showed that the tolerance to both recipient-type and donor-type major histocompatibility complex determinants was induced in the chimeric mice. In the peripheral blood (PB) of these mice, only one type of cells from the three different donor strains became dominant in most chimeric mice and reached a stable level about 4 months after BMT. Polymerase chain reaction analyses, however, revealed that the skins from all the donors were accepted even when no cells with their phenotypes could be detected in the PB. These results suggest that both hemato-lymphoid reconstitution and stable tolerance to not only the recipient strain but also all the donor strains can be achieved in chimeric mice, indicating the possibility of mixed CB transplantation in humans. PMID- 15342934 TI - Elevation of platelet activation markers and chemokines during peripheral blood stem cell harvest with G-CSF. AB - The kinetics of peripheral blood stem cell mobilization in response to recombinant human granulocyte colony-stimulating factor is well established. However, there have been few investigations of platelet activation markers during peripheral blood stem cell harvest. We measured the levels of the platelet activation markers, chemokines, and soluble factors in plasma obtained from patients undergoing peripheral blood stem cell harvest. The number of leukocytes, CD34+ cells, neutrophils, monocytes, and lymphocytes peaked on day 5 after granulocyte colony-stimulating factor treatment, but the numbers of eosinophils and basophils showed no significant change. Regulated on activation normally T cell expressed and secreted (RANTES) level increased through day 10, and the monocyte chemotactic peptide-1 (MCP-1) level peaked on day 5. Platelet counts continued to increase through day 10. The level of thrombopoietin significantly increased on day 3, peaked on day 5, and decreased slightly by day 10. The levels of soluble CD40 ligand and soluble P-selectin increased up to day 5. The platelet derived microparticle level peaked on day 5, and then began to decline. CD34+ cell numbers significantly correlated with those of leucocytes, neutrophils, monocytes, and lymphocytes, as well as levels of MCP-1, and the CD34+ cells exhibited changes similar to platelet-derived microparticles. The patterns of change in MCP-1, platelet-derived microparticles, and the CD34+ cell count are similar in that each peaks on day 5 and decreases thereafter. Further study is required to determine if a cause-and-effect relationship in their pattern of change exists among them. PMID- 15342935 TI - AC133+ G0 cells from cord blood show a high incidence of long-term culture initiating cells and a capacity for more than 100 million-fold amplification of colony-forming cells in vitro. AB - AC133+ cells may provide an alternative to CD34+ cells as a target for cell expansion and gene therapy protocols. We examined the differences in proliferative potential between cord blood selected for AC133 or CD34 in serum free, stroma cell-free culture for up to 30 weeks. Because most hemopoietic stem cells reside within the G0/G1 phase of the cell cycle, we combined enrichment according to AC133 or CD34 expression with G0 position in the cell cycle to identify populations enriched for putative stem cells. Our results show that AC133+ G0 cells demonstrated a long-term culture-initiating cell incidence of 1 in 4.2 cells, had a colony-forming cell incidence of 1 in 2.8 cells, were capable of producing 660 million-fold expansion of nucleated cells and 120 million-fold expansion of colony-forming units-granulocyte-macrophage over a period of 30 weeks, and were consistently superior to CD34+ G0 cells according to these parameters. Furthermore, we have shown that AC133+CD34- cells have the ability to generate CD34+ cells in culture, which suggests that at least some AC133+ cells are ancestral to CD34+ cells. We conclude that AC133 isolation provides a better means of selection for primitive hemopoietic cells than CD34 and that, in combination with isolation according to G0 phase of the cell cycle, AC133 isolation identifies a highly enriched population of putative stem cells. PMID- 15342936 TI - Simultaneous maintenance of human cord blood SCID-repopulating cells and expansion of committed progenitors at low O2 concentration (3%). AB - In the present work, we tested the hypothesis that liquid cultures (LCs) of cord blood CD34+ cells at an appropriate low O2 concentration could simultaneously allow colony-forming cell (CFC) expansion and nonobese diabetic/severe combined immunodeficiency mice-repopulating cell (SRC) maintenance. We first found that 3% was the minimal O2 concentration, still allowing the same rate of CFC expansion as at 20% O2. We report here that 7-day LCs of cord blood CD34+ cells at 3% O2 maintain SRC better than at 20% O2 and allow a similar amplification of CFCs (35- to 50-fold) without modifying the CD34+ cell proliferation. Their phenotypic profile (antigens: HLA-DR, CD117, CD33, CD13, CD11b, CD14, CD15, and CD38) was not modified, with exception of CD133, whose expression was lower at 3% O2. These results suggest that low O2 concentrations similar to those found in bone marrow participates in the regulation of hematopoiesis by favoring stem cell-renewing divisions. This expansion method that avoids stem cell exhaustion could be of paramount interest in hematopoietic transplantation by allowing the use of small size grafts in adults. PMID- 15342937 TI - Selective generation of different dendritic cell precursors from CD34+ cells by interleukin-6 and interleukin-3. AB - There is a growing interest in generating dendritic cells (DCs) for using as vaccines. Several cytokines, especially stem cell factor (SCF) and FLT3-ligand (FL), have been identified as essential to produce large numbers of myeloid precursors and even to increase DC yield obtained by the action of granulocyte macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor alpha (TNF-alpha). However, there are few studies on the effect of the early-acting cytokines, commonly used to expand CD34+ progenitor cells, on DC generation. We report here that in the absence of serum, SCF, FL, and thrombopoietin (TPO) plus interleukin-6 (IL-6) and SCF, FL, and TPO plus IL-3 were able to generate CD14+CD1a- and CD14- CD1a+ myeloid DC precursors from CD34+ cells, but IL-6 had an inhibitory effect on the generation of CD14- CD1a+ cells. Both DC precursors differentiated into mature DCs by GM-CSF, IL-4, and TNF-alpha, and DCs obtained from both types of culture exhibited equal allostimulatory capacity. CD1a+ DCs generated could be identified on the basis of DC-specific intracellular adhesion molecule-grabbing nonintegrin (DC-SIGN) expression, a novel C-type lectin receptor expressed on dermal DCs but not on Langerhans cells. In addition, the inclusion of IL-3 to the culture medium induced the appearance of CD13- cells that differentiated into plasmacytoid DC (DC2) on the addition of TNF-alpha, allowing the identification of developmental stages of DC2. Like true plasmacytoid DCs, these cells secreted interferon-alpha after TLR9-specific stimulation with a specific CpG nucleotide. PMID- 15342938 TI - Effects of telomerase modulation in human hematopoietic progenitor cells. AB - Loss of telomeric repeats has been causally linked to replicative senescence and aging in human cells. In contrast to normal somatic cells, which are telomerase negative, hematopoietic stem cells have low levels of telomerase, which can be transiently upregulated upon cytokine stimulation. To examine whether ectopic expression of telomerase can overcome telomere erosion in hematopoietic progenitor cells, we overexpressed telomerase in CD34+ and AC133+ cord blood (CB) cells using retroviral vectors containing hTERT, the catalytic component of telomerase. Although the hTERT-transduced CB cells exhibited significantly elevated telomerase activity (approximately 10-fold), the mean telomere length was only increased up to 600 bp, which was in contrast to hTERT-transduced fibroblast cells gaining more than 2-kb telomeric repeats. Moreover, ectopic telomerase activity did not prevent overall telomere shortening, which was in the range of 1.3 kb in serum-free expansion culture. We also blocked endogenous telomerase activity by ectopic expression of dominant-negative hTERT. Whereas CB cells with absent telomerase activity showed reduced absolute numbers of colony forming cells, we observed increased rates only for burst-forming units erythroid when the enzyme was overexpressed. These results suggest that telomere shortening in human hematopoietic progenitor cells cannot be compensated by increased levels of telomerase alone and is likely to be dependent on other factors, such as telomere binding proteins. Furthermore, telomerase function seems to be directly associated with the proliferative capacity of stem cells and may exert an additional role in lineage differentiation. PMID- 15342939 TI - Exposure of hematopoietic stem cells to benzene or 1,4-benzoquinone induces gender-specific gene expression. AB - Chronic exposure to benzene results in progressive decline of hematopoietic function and may lead to the onset of various disorders, including aplastic anemia, myelodysplastic syndrome, and leukemia. Damage to macromolecules resulting from benzene metabolites and misrepair of DNA lesions may lead to changes in hematopoietic stem cells (HSCs) that give rise to leukemic clones. We have shown previously that male mice exposed to benzene by inhalation were significantly more susceptible to benzene-induced toxicities than females. Because HSCs are targets for benzene-induced cytotoxicity and genotoxicity, we investigated DNA damage responses in HSC from both genders of 129/SvJ mice after exposure to 1,4-benzoquinone (BQ) in vitro or benzene in vivo. 1,4-BQ is a highly reactive metabolite of benzene that can cause cellular damage by forming protein and DNA adducts and producing reactive oxygen species. HSCs cultured in the presence of 1,4-BQ for 24 hours showed a gender-independent, dose-dependent cytotoxic response. RNA isolated from 1,4-BQ-treated HSCs and HSCs from mice exposed to 100 ppm benzene by inhalation showed altered expression of apoptosis, DNA repair, cell cycle, and growth control genes compared with unexposed HSCs. Rad51, xpc, and mdm-2 transcript levels were increased in male but not female HSCs exposed to 1,4-BQ. Males exposed to benzene exhibited higher mRNA levels for xpc, ku80, ccng, and wig1. These gene expression differences may partially explain the gender disparity in benzene susceptibility. HSC culture systems such as the one used here will be useful for testing the hematotoxicity of various substances, including other benzene metabolites. PMID- 15342940 TI - Fetal immune suppression as adjunctive therapy for in utero hematopoietic stem cell transplantation in nonhuman primates. AB - In utero hematopoietic stem cell transplantation could potentially be used to treat many genetic diseases but rarely has been successful except in severe immunodeficiency syndromes. We explored two ways to potentially increase chimerism in a nonhuman primate model: (a) fetal immune suppression at the time of transplantation and (b) postnatal donor stem cell infusion. Fetal Macaca nemestrina treated with a combination of the corticosteroid betamethasone (0.9 mg/kg) and rabbit thymoglobulin (ATG; 50 mg/kg) were given haploidentical, marrow derived, CD34+ -enriched donor cells. Animals treated postnatally received either donor-derived T cell-depleted or CD34+ -enriched marrow cells. Chimerism was determined by traditional and real-time polymerase chain reaction from marrow, marrow progenitors, peripheral blood, and mature peripheral blood progeny. After birth, the level of chimerism in the progenitor population was higher in the immune-suppressed animals relative to controls (11.3% +/- 2.7% and 5.1% +/- 1.5%, respectively; p = .057). Chimerism remained significantly elevated in both marrow (p = .02) and fluorescence-activated cell sorted and purified CD34+ cells (p = .01) relative to control animals at > or = 14 months of age. Peripheral blood chimerism, both at birth and long term, was similar in immune-suppressed and control animals. In the animals receiving postnatal donor cell infusions, there was an initial increase in progenitor chimerism; however, at 6-month follow-up, the level of chimerism was unchanged from the preinfusion values. Although fetal immune suppression was associated with an increase in the level of progenitor and marrow chimerism, the total contribution to marrow and the levels of mature donor progeny in the peripheral blood remained low. The level of long-term chimerism also was not improved with postnatal donor cell infusion. PMID- 15342941 TI - LIF/STAT3 signaling fails to maintain self-renewal of human embryonic stem cells. AB - Murine embryonic stem (mES) cells remain undifferentiated in the presence of leukemia inhibitory factor (LIF), and activation of signal transducer and activator of transcription 3 (STAT3) via LIF receptor (LIFR) signaling appears sufficient for maintenance of mES cell pluripotency. Anecdotal and contradictory accounts exist for the action of LIF in the culture of human embryonic stem cells, and the nature of LIF signaling and whether the LIF-STAT3 pathway is conserved in human embryonic stem cells (hESCs) has not been systematically explored. In this study, we show that the LIFRbeta and the signaling subunit gp130 are expressed in hESCs and that human LIF can induce STAT3 phosphorylation and nuclear translocation in hESCs. Nevertheless, despite the functional activation of the LIF-STAT3 signaling pathway, human LIF is unable to maintain the pluripotent state of hESCs. Feeder-free culture conditions that maintain hESCs in an undifferentiated state do not show activation of STAT3, suggesting that distinct signaling mechanisms govern the self-renewal of hESCs. PMID- 15342942 TI - An efficient and safe xeno-free cryopreservation method for the storage of human embryonic stem cells. AB - Human embryonic stem cells (hESCs) promise to revolutionize reparative medicine through their potential in developing cell replacement therapies for diseases like diabetes and parkinsonism. Most of the existing hESC lines available for research, including all National Institutes of Health-registered lines, have been derived and maintained on mouse embryonic fibroblast feeders in the presence of xenoproteins. For future clinical application, many more hESC lines derived and grown in current good manufacturing practice, good tissue culture practice, and xeno-free conditions need to be developed. Concurrently, effective cryopreservation methods that prevent or limit the accidental contact of hESCs with nonsterile liquid nitrogen during periods of long-term storage have to be formulated. We describe a safe, xeno-free cryopreservation protocol for hESCs involving vitrification in closed sealed straws using human serum albumin as opposed to fetal calf serum as the main protein source in the cryoprotectant and long-term storage in the vapor phase of liquid nitrogen. After thaw, hESCs exhibited high thaw-survival rates and low differentiation rates, remained pluripotent, and maintained normal diploid karyotypes throughout extended passage. The cryopreservation technique we describe here should complement xeno free culture conditions for hESCs already in refinement and will prove very useful for the setting up of hESC banks throughout the world. PMID- 15342943 TI - Derivation of human embryonic stem cells from day-8 blastocysts recovered after three-step in vitro culture. AB - Human embryonic stem cells (hESCs) have been derived from the inner cell mass (ICM) of day 5-7 blastocysts and hold great promise for research into human developmental biology and the development of cell therapies for the treatment of human diseases. We report here that our novel three-step culture conditions successfully support the development of day-8 human blastocysts, which possess significantly (p <.01) more ICM cells than day-6 blastocysts. Plating of ICMs isolated from day-8 blastocysts resulted in the formation of a colony with hESC morphology from which a new hESC line (hES-NCL1) was derived. Our stem cell line is characterized by the expression of specific cell surface and gene markers: GTCM-2, TG343, TRA1-60, SSEA-4, alkaline phosphatase, OCT-4, NANOG, and REX-1. Cytogenetic analysis of the hESCs revealed that hES-NCL1 line has a normal female (46, XX) karyotype. The pluripotency of the cell line was confirmed by the formation of teratomas after injection into severely combined immunodeficient mice and spontaneous differentiation under in vitro conditions. PMID- 15342944 TI - Requirement for neurogenesis to proceed through the division of neuronal progenitors following differentiation of epidermal growth factor and fibroblast growth factor-2-responsive human neural stem cells. AB - Epidermal growth factor (EGF)- and fibroblast growth factor-2 (FGF-2)-responsive human neural stem cells may provide insight into mechanisms of neural development and have applications in cell-based therapeutics for neurological disease. However, their biology after expansion in vitro is currently poorly understood. Cells grown in either EGF or FGF-2 or a combination of both mitogens displayed characteristically similar levels of transcriptional activation and comparable proliferative profiles with linear cell-cycle kinetics and possessed similar neuronal differentiation capabilities. These data support the view that human neurospheres at later stages of expansion (>10 weeks) are comprised overwhelmingly of a single type of stem cell responsive to both EGF and FGF-2. After mitogen withdrawal and neurosphere plating, bromodeoxyuridine pulse-chase experiments revealed that the stem cells did not undergo differentiation directly into neurons. Instead, most immature neurons arose via the division of emerging progenitor cells in the absence of exogenous EGF or FGF-2. Neurogenesis was abolished by application of high concentrations of either EGF/FGF-2 or the mitotic inhibitor cytosine-b-arabinofuranoside, suggesting that there is an obligatory requirement for at least one round of cell division in the absence of mitogens as a prelude to terminal neuronal differentiation. The differentiation of human neurospheres provides a useful model of human neurogenesis, and the data presented indicate that it proceeds through the division of committed neuronal progenitor cells rather than directly from the neural stem cell. PMID- 15342945 TI - Contribution of bone marrow-derived cells to skin: collagen deposition and wound repair. AB - The bone marrow provides inflammatory cells and endothelial progenitor cells to healing cutaneous wounds. To further explore the bone marrow contribution to skin and healing wounds, we used a chimeric mouse model in which the bone marrow from enhanced green fluorescent protein (EGFP) transgenic mice is transplanted into normal C57BL mice. We found that normal skin is a target organ for bone marrow derived cells from both the hematopoietic and the mesenchymal stem cell pool. We present evidence that the bone marrow contribution to normal skin and the healing cutaneous wound is substantially greater than the previously recognized CD45+ subpopulation, where 15%-20% of the spindle-shaped dermal fibroblasts were bone marrow-derived (EGFP+). Furthermore, the bone marrow-derived cells were able to contract a collagen matrix and transcribe both collagen types I and III, whereas the skin-resident cells transcribed only collagen type I. Whereas endothelial progenitor cells were found early during the wound repair process, bone marrow derived endothelial cells were not seen after epithelialization was complete. Our data show that wound healing involves local cutaneous cells for reconstituting the epidermis but distant bone marrow-derived cells and the adjacent uninjured dermal mesenchymal cells for reconstituting the dermal fibroblast population. PMID- 15342946 TI - Isolation of a highly clonogenic and multipotential subfraction of adult stem cells from bone marrow stroma. AB - Attempts have been made to develop cell and gene therapies using the adult stem cells from bone marrow referred to as mesenchymal stem cells or marrow stromal cells (MSCs). However, the results have been variable in part because there are no standardized protocols for preparing and characterizing MSCs. In the experiments presented here, we developed a standardized assay by light scattering to measure the content of rapidly self-renewing cells (RS cells) in preparations of MSCs. The assay quickly identifies preparations of MSCs that replicate rapidly in subsequent culture. In addition, the standardized assay enabled us to isolate RS cells that were up to 90% clonogenic and that generated single cell-derived colonies that differentiated into either mineralizing cells or adipocytes with appropriate additions to the medium. PMID- 15342947 TI - Inhibition of growth by p205: a nuclear protein and putative tumor suppressor expressed during myeloid cell differentiation. AB - p205 belongs to a family of interferon-inducible proteins called the IFI-200 family, which have been implicated in the regulation of cell growth and differentiation. While p205 is induced in hematopoietic stem cells during myeloid cell differentiation, its function is not known. Therefore, the aim of this study was to determine the role of p205 in regulating proliferation in hematopoietic progenitor cells and in nonhematopoietic cell lines. We found that p205 localizes to the nucleus in hematopoietic and nonhematopoietic cell lines. Transient expression of p205 in murine IL-3-dependent BaF3 and 32D-C123 progenitor cell lines inhibited IL-3-induced growth and proliferation. The closely related IFI 200 family members, p204 and p202, similarly inhibited IL-3-dependent progenitor cell proliferation. p205 also inhibited the proliferation and growth of normal hematopoietic progenitor cells. In nonhematopoietic cell lines, p205 and p204 expression inhibited NIH3T3 cell colony formation in vitro, and microinjection of p205 expression vectors into NIH3T3 fibroblasts inhibited serum-induced proliferation. We have determined the functional domains of p205 necessary for activity, which were identified as the N-terminal domain in apoptosis and interferon response (DAPIN)/PYRIN domain, and the C-terminal retinoblastoma protein (Rb)-binding motif. In addition, we have demonstrated that a putative ataxia telangiectasia, mutated (ATM) kinase phosphorylation site specifically regulates the activity of p205. Taken together, these data suggest that p205 is a potent cell growth regulator whose activity is mediated by its protein-binding domains. We propose that during myelomonocytic cell differentiation, induction of p205 expression contributes to cell growth arrest, thus allowing progenitor cells to differentiate. PMID- 15342948 TI - Expression profiling and functional analysis of wnt signaling mechanisms in mesenchymal stem cells. AB - Through their broad differentiation potential, mesenchymal stem cells (MSCs) are candidates for a range of therapeutic applications, but the precise signaling pathways that determine their differentiated fate are not fully understood. Evidence is emerging that developmental signaling cues may be important in regulating stem cell self-renewal and differentiation programs. Here we have identified a consistent expression profile of Wnt signaling molecules in MSCs and provide evidence that an endogenous canonical Wnt pathway functions in these cells. Wnts bind to Frizzled (Fz) receptors and subsequent canonical signaling inhibits glycogen synthase kinase-3beta (GSK-3beta), causing beta-catenin translocation into the nucleus to induce target gene expression. In human MSCs isolated from bone marrow of different donors, we appear to have identified a common Wnt/Fz expression profile using reverse transcriptase polymerase chain reaction (RT-PCR). Associated Wnt signaling components, including low-density lipoprotein receptor-related protein-5 (LRP-5), kremen-1, dickkopf-1 (Dkk-1), secreted Frizzled-related peptide (sFRP)-2, sFRP3, sFRP4, Disheveled (Dvl), GSK 3beta, adenomatous polyposis coli (APC), beta-catenin,T-cell factor (TCF)-1, and TCF-4, were also identified. Nuclear beta-catenin was observed in 30%-40% of MSCs, indicative of endogenous Wnt signaling. Exposure to both Wnt3a and Li+ ions, which promotes canonical Wnt signaling by inhibiting GSK-3beta, reduced phosphorylation of beta-catenin in MSCs and increased beta-catenin nuclear translocation approximately threefold over that of the controls. Our findings indicate that autocrine Wnt signaling operates in primitive MSC populations and supports previous evidence that Wnt signaling regulates mesenchymal lineage specification. The identification of a putative common Wnt/Fz molecular signature in MSCs will contribute to our understanding of the molecular mechanisms that regulate self-renewal and lineage-specific differentiation. PMID- 15342949 TI - STAT3 is dispensable for maintenance of self-renewal in nonhuman primate embryonic stem cells. AB - The leukemia inhibitory factor (LIF)/glycoprotein 130 (gp130)/signal transducer and activator of transcription 3 (STAT3) pathway plays an essential role in the maintenance of self-renewal and pluripotency in mouse embryonic stem (ES) cells. However, in primate ES cells, including those from humans and monkeys, LIF alone is not sufficient to maintain self-renewal. The precise role of the LIF/gp130/STAT3 pathway for self-renewal in primate ES cells is still unclear. In this study, we found that stimulation of cynomolgus monkey ES cells with LIF or interleukin (IL)-6/soluble IL-6 receptor leads to STAT3 phosphorylation, an effect seen previously in murine ES cells. Concomitant with this notion, nuclear translocalization and transcriptional activation of STAT3 were observed in a LIF dependent manner. Moreover, the analysis of a dominant interfering mutant, STAT3F, showed that even though the phosphorylation, nuclear translocalization, and transcriptional activation of endogenous STAT3 after LIF stimulation were completely abrogated by over-expressing STAT3F in monkey ES cells, they continued to proliferate in an undifferentiated state, retaining their pluripotency. These results demonstrate that the LIF/gp130/STAT3 pathway functions in cynomolgus monkey ES cells but is not essential for the maintenance of self-renewal. They also suggest that cynomolgus monkey ES cells, unlike murine ES cells, are maintained in an undifferentiated state through LIF/gp130/STAT3-independent signaling. PMID- 15342950 TI - The molecular perspective: polycyclic aromatic hydrocarbons. PMID- 15342952 TI - A toxicogenomic approach to drug-induced phospholipidosis: analysis of its induction mechanism and establishment of a novel in vitro screening system. AB - Phospholipidosis is a lipid storage disorder in which excess phospholipids accumulate within cells. Some cationic amphiphilic compounds are known to have the potential to induce phospholipidosis. This study was undertaken to examine the molecular mechanisms that contribute to the development of phospholipidosis and to identify specific markers that might form the basis of an in vitro screening test. Specifically, we performed a large-scale gene expression analysis using DNA microarrays on human hepatoma HepG2 cells after they were treated with each of 12 compounds known to induce phospholipidosis. In electron microscopy, HepG2 cells developed lamellar myelin-like bodies in their lysosomes, the characteristic change of phospholipidosis, after treatment with these compounds for 72 h. DNA microarray analysis performed 6 and 24 h after treatment showed alterations in gene expression reflecting the inhibition of lysosomal phospholipase activity and lysosomal enzyme transport, and the induction of phospholipid and cholesterol biosynthesis. Seventeen genes that showed a similar expression profile following treatment were selected as candidate markers. Real time PCR analysis confirmed that 12 gene markers showed significant concordance with lamellar myelin-like body formation. Furthermore, the average fold change values of these markers correlated well with the magnitude of this pathological change. In conclusion, microarray analysis revealed that factors such as alterations in lysosomal function and cholesterol metabolism were involved in the induction of phospholipidosis. Furthermore, comprehensive gene expression analysis enabled us to identify biomarkers of this condition that we then used to develop a rapid and sensitive in vitro screening test for drug-induced phospholipidosis. PMID- 15342953 TI - Arsenic speciation in bile and urine following oral and intravenous exposure to inorganic and organic arsenics in rats. AB - Although inorganic arsenate (iAsV) and arsenite (iAsIII) are metabolized in liver and excreted into bile and urine, the metabolites in the bile after the oral intake of iAs remain unclear. Male Sprague-Dawley rats were orally (po) or intravenously (iv) exposed to iAs and methylated arsenics, and the arsenic speciation in the urine and bile was analyzed by high performance liquid chromatography-inductively coupled argon plasma mass spectrometry. Arsenic caused induction of multidrug resistance-associated protein 2 (MRP2), and changes of glutathione (GSH) levels in the liver and bile were also determined. The metabolic speciation studies revealed that arsenic was excreted into bile in the methylarsenic-diglutathione (MADG) and/or dimethylarsenic acid (DMAV) forms in iAsIII- or iAsV-po rats, but that MADG and arsenic-triglutathione (ATG) are the main forms excreted into bile both in iAsIII- and iAsV-iv rats. In MADG-po rats, the MADG was excreted into bile in the MADG and DMAV forms. Monomethylarsonic acid (MMAV)- and DMAV-iv rats did not excrete significant amounts of either MMAV or DMAV into bile and mostly excreted into urine in the unchanged chemical forms. Taken together, the DMAV detected in the bile is mostly supposed to be the dissociation of dimethylarsenic-glutathione (DMAG). Urinary arsenic speciation showed that arsenic metabolized to 43% methylated DMAV, 47% unmethylated iAsIII, and 10% iAsV in iAsIII-iv rats, whereas only 3% methylated DMAV, 87% unmethylated iAsV, and 10% iAsIII were detected in iAsV-iv rats. Arsenic was accumulated dose dependently, and arsenic concentration was significantly higher in the iAsIII-po rat liver than in the iAsV-po rat liver. GSH levels in the bile were decreased by relatively higher doses of iAsV-po, but significantly increased by iAsIII- or iAsV-iv. iAs-exposure increased the expression of MRP2 in the liver. Pretreatment with buthionine sulfoximine predominantly inhibited arsenic excretion into bile in iAs-iv rats. In conclusion, our data demonstrated that biliary and urinary arsenic excretion and speciation are affected by the route, dose, and chemical forms of arsenical administration, and GSH plays a key role in arsenic metabolism. We are also first to show that DMAV that probably originated from DMAG is excreted into the bile in iAs-po rats. PMID- 15342954 TI - Effect of dibromoacetic acid on DNA methylation, glycogen accumulation, and peroxisome proliferation in mouse and rat liver. AB - Dibromoacetic acid (DBA) is a drinking water disinfection by-product. Its analogs, dichloroacetic acid (DCA) and trichloroacetic acid (TCA), are liver carcinogens in rodents. We evaluated the ability of DBA to cause DNA hypomethylation, glycogen accumulation, and peroxisome proliferation that are activities previously reported for the two other haloacetic acids. Female B6C3F1 mice and male Fischer 344 rats were administered 0, 1,000, and 2,000 mg/l DBA in drinking water. The animals were euthanized after 2, 4, 7, and 28 days of exposure. Dibromoacetic acid caused a dose-dependent and time-dependent decrease of 20%-46% in the 5-methylcytosine content of DNA. Hypomethylation of the c-myc gene was observed in mice after 7 days of DBA exposure. Methylation of 24 CpG sites in the insulin-like growth factor 2 (IGF-II) gene was reduced from 80.2% +/ 9.2% to 18.8% +/- 12.9% by 2,000 mg/l DBA for 28 days. mRNA expression of the c myc and IGF-II genes in mouse liver was increased by DBA. A dose-dependent increase in the mRNA expression of the c-myc gene was also observed in rats. In both mice and rats, DBA caused dose-dependent accumulation of glycogen and an increase of peroxisomal lauroyl-CoA oxidase activity. Hence, DBA, like DCA and TCA, induced hypomethylation of DNA and of the c-myc and IGF-II genes, increased mRNA expression of both genes, and caused peroxisome proliferation. Again like DCA, DBA also induced glycogen accumulation. These results indicate that DBA shares biochemical and molecular activities in common with DCA and/or TCA, suggesting that it might also be a liver carcinogen. PMID- 15342955 TI - The cyanobacterial toxin cylindrospermopsin inhibits pyrimidine nucleotide synthesis and alters cholesterol distribution in mice. AB - The hepatotoxin Cylindrospermopsin, a sulfated-guanidinium alkaloid with substituted dioxypyrimidine (uracil) moiety, was isolated from several cyanobacteria species. Our previous studies on the toxicity of cylindrospermopsin and its derivatives suggested that the uracil moiety is crucial for the toxicity and that such toxicity could partly stem from competitive binding of the toxin to a catalytic site(s) involved in the synthesis of pyrimidine nucleotides (i.e., uridine). In the present study we demonstrated that cylindrospermopsin inhibited in a noncompetitive manner the in vitro activity of uridine monophosphate (UMP) synthase complex (responsible for the conversion of orotic acid to UMP) in a cell free liver extract from mice, with an inhibition constant, KI, of 10 microM. Exposure of mice to cylindrospermopsin at subacute concentrations, via drinking water, only slightly affected the in vitro activity of UMP synthase. The typical metabolic disorder associated with the inhibition of UMP synthase activity, known as "orotic aciduria," was not observed under these conditions, but other anomalous metabolic responses related to cholesterol metabolism were developed. PMID- 15342956 TI - Analyses of glutathione reductase hypomorphic mice indicate a genetic knockout. AB - A strain of mice (Gr1a1Neu) that exhibited tissue glutathione reductase (GR) activities that were substantially lower (less than 10% in liver) than the corresponding activities in control mice has been reported. The present report describes characterization of the mutation(s) in the GR gene of these mice. RT PCR of mRNA from the Neu mice indicated a substantial deletion in the normal GR coding sequence. Southern blots revealed that the deletion involved a region spanning from intron 1 through intron 5. The exact breakpoints of the deletion were characterized by PCR and sequencing through the region encompassing the deletion. The deletion involves nucleotides 10840 through 23627 of the genomic GR gene and functionally deletes exons 2 through 5. In addition, the deletion produces a frame shift in exon 6 and introduces a stop codon in exon 7 that would prevent translation of the remainder of the protein. Consequently, the Neu mice are incapable of producing a functional GR protein and appear to be genetic knockouts for GR. The Neu mice offer live animal models with which to test hypotheses regarding oxidant mechanisms of tissue injury in vivo. PMID- 15342957 TI - Block of neuronal nicotinic acetylcholine receptors by organophosphate insecticides. AB - Chronic and acute exposure to organophosphate (OP) pesticides may lead to persistent neurological and neurobehavioral effects, which cannot be explained by acetylcholinesterase (AChE) inhibition alone. It is suggested that other brain proteins are involved. Effects of commonly used organophosphate pesticides on rat neuronal alpha4beta2 nicotinic acetylcholine receptors (nAChRs) expressed in Xenopus laevis oocytes have been investigated using the two-electrode voltage clamp technique. Several OP pesticides, e.g., parathion-ethyl, chlorpyrifos and disulfoton, inhibited the ACh-induced ion current with potencies in the micromolar range. The potency of inhibition increased with increasing concentrations of the agonist ACh. Comparison of the potency of nAChR inhibition with the potency of AChE inhibition demonstrated that some OPs inhibit nAChRs more potently than AChE. Binding experiments on alpha4beta2 nAChRs showed that the OPs noncompetitively interact with nAChRs. The inhibitory effects on nAChRs are adequately described and explained by a sequential two-step mechanism, in which rapidly reversible OP binding to a separate binding site leads to inhibition followed by a stabilization of the blocked state or receptor desensitization. It is concluded that OPs interact directly with neuronal alpha4beta2 nAChRs to inhibit the agonist-induced response. This implicates that neuronal alpha4beta2 nAChRs are additional targets for some OP pesticides. PMID- 15342958 TI - Functional innervation in tissue engineered models for in vitro study and testing purposes. AB - The biotechnology industry is rapidly expanding and the emerging field of tissue engineering is projected to have a high impact in the near future. Recently the field of cellular, drug, and prosthetic delivery has melded with the field of tissue engineering to make simulated tissues. In addition to their roles as tissue substitutes for transplantation, these simulated tissues may provide more accurate models and environments for toxicology testing and the study of peripheral nerves. The current study demonstrates the importance of innervation, in general, for the function of engineered tissues. We observe that the presence of nerves in a tissue engineered (TE) human cornea model enhances the growth of the epithelium and the formation of its protective mucin layer. Innervation also confers protection to the epithelium from chemical insult, as determined by the level of post-treatment epithelial cell death. We demonstrate differential responses of the nerves to chemical stimuli by changes in intracellular sodium as measured by 2-photon microscopy. The 2-photon imaging techniques also allow for the visualization and study of the fine sensory axon fibers within the 3 dimensional tissue. This work demonstrates a role for innervation in the protective quality and function of the engineered tissue, and the potential to use the nerves themselves as indicators of the severity of an insult. These results are important to consider for the development of any optimized TE models for in vitro study and testing purposes. PMID- 15342959 TI - The accuracy of extended histopathology to detect immunotoxic chemicals. AB - The accuracy of extended histopathology to detect immunotoxic chemicals in female B6C3F1 mice was evaluated under the auspices of the National Toxicology Program (NTP). A workgroup was formed consisting of four pathologists who conducted extended histopathological evaluation of lymphoid tissues obtained from a subset of NTP toxicology studies, in which previously detailed immunotoxicity assessment was performed. In addition, a positive control data set of three known immunosuppressive agents, one negative control data set, and an additional negative control group composed of the vehicle only treated groups were included. Data obtained from extended histopathology evaluations were compared to more traditional immune test results (both functional and nonfunctional) from previously conducted immunotoxicity assessments. Analyses of the data indicated that the ability to identify immunotoxic chemicals using histological endpoints decreased linearly as the level of stringency used to determine significant histopathological changes increased. A relatively high (80%) accuracy level was achieved when histological changes were considered in toto (i.e., any histological abnormality in the three tissues examined), using minimal or mild criteria for scoring. When minimal or mild histological changes were considered significant for a specific tissue, a 60% level of accuracy in identifying immunotoxic chemicals was obtained as compared to a 90% accuracy level that was achieved with this data set using the antibody plaque forming cell response, considered to represent the most predictive functional test. A minimal classification was obtained in the analyses of the negative control groups, suggesting that use of the minimal classification for hazard identification is inappropriate as it will likely result in a high incidence of false positives. This was not the case when mild classifications were used as an indicator of significance, which in most instances allowed the successful identification of negatives. When moderate to marked histopathological changes were used to identify immunotoxic chemicals, the level of accuracy that could be achieved was poor. A considerably higher level of accuracy was obtained for the positive control data set than the test chemical data set suggesting that the ability to detect an immunotoxic agent histologically is proportional to the potency of the immunotoxic agent. Comparison of immune function test results and histopathological results obtained from the high-dose treatment groups and the lower-dose treatment group did not reveal any significant differences between the two endpoints to predict immunotoxicity as a function of dose. Of the three lymphoid organs examined, (i.e., lymph node, thymus, and spleen), the most consistent and discernible histological lesions were observed in the thymus cortical region. These lesions correlated with thymus: body weight ratios and to a slightly lesser extent, the antibody plaque forming cell response. Addition of general toxicological endpoints such as body weight and leukocyte counts did not significantly improve the sensitivity of extended histopathology for this data set. Taken together, these data suggest that, while not as sensitive as functional analyses, extended histopathology may provide a reasonable level of accuracy as a screening test to identify immunotoxic chemicals, provided the level of stringency used to score histological lesions is carefully considered to allow for detection of immunotoxic agents while limiting false positives. PMID- 15342960 TI - EGR1 is a novel target for AhR agonists in human lung epithelial cells. AB - The transcription factor early growth response 1 (EGR1) was previously identified as a potential novel target of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in human lung epithelial cells by toxicogenomic analysis. EGR1 has been implicated in the pathogenesis of vascular disease and is altered by a number of factors that include stress, inflammation, and hypoxia. Depending on its downstream targets or protein interactions, EGR1 regulates important biological processes that include cell growth, apoptosis, and differentiation. The following experiments were conducted to determine if EGR1 is indeed a target of TCDD and polycyclic aromatic hydrocarbons (PAHs) that can act through a similar mechanism. Pulmonary epithelial cells were exposed to TCDD for 24 h and an increase in EGR1 mRNA was measured. In addition, EGR1 protein was increased by TCDD and PAHs that have binding affinity to the aryl hydrocarbon receptor. The transcriptional activity of the EGR1 promoter was measured with a luciferase construct; however, no increases in luciferase activity were detected in TCDD or PAH-treated cells. Using actinomycin to inhibit RNA synthesis, we found that TCDD increased the half life of EGR1 mRNA from 13 to 22 min. Thus, the increase in EGR1 expression appears to be mediated through a post-transcriptional mechanism that leads to the higher EGR1 protein levels in TCDD and PAH treated cells, compared to vehicle treated cells. Increased expression of a transcription factor EGR1 with tumorigenic and other biological activities could contribute to the deleterious pulmonary effects of exposure to these environmental agents. PMID- 15342961 TI - Toxin-induced tail phosphorylation of hepatocellular S6 kinase: evidence for a dual involvement of the AMP-activated protein kinase in S6 kinase regulation. AB - Several protein phosphatase-inhibitory toxins (okadaic acid, microcystin, calyculin A, cantharidin, tautomycin) administered to isolated rat hepatocytes were found to induce phosphorylation in the tail region of S6 kinase (S6K; p70S6K1) as detected with a phosphospecific antibody against doubly phosphorylated Thr-421/Ser424. 5-Aminoimidazole-4-carboxamide riboside (AICAR), an adenosine analogue that elicits activation of the hepatocellular AMP-activated protein kinase (AMPK), similarly stimulated S6K tail phosphorylation. The flavonoid naringin prevented the effects of AICAR, okadaic acid, and microcystin on AMPK activation as well as on S6K tail phosphorylation, suggesting AMPK as a mediator of the latter. The effects of AICAR and the toxins were rapamycin resistant; in contrast, amino acids induced an S6K tail phosphorylation that was rapamycin sensitive, suggesting mediation by the protein kinase mammalian target of rapamycin (mTOR). Amino acids activated S6K by phosphorylation at Thr-389, but the toxins did not, and AICAR in fact suppressed the activating phosphorylation induced by the amino acids. The possibility thus must be considered that the phosphorylated S6K tail may transmit a toxin-induced signal independently of S6K enzymatic activity. Despite their inability to activate S6K, the toxins (but not AICAR) stimulated phosphorylation of the ribosomal protein S6, presumably by activating some other S6-phosphorylating protein kinase. PMID- 15342962 TI - Morphological and biochemical characterization of macrophages activated by carrageenan and lipopolysaccharide in vivo. AB - Macrophages are able to recognize, internalize and destroy a large number of pathogens, thus restricting the infection until adaptive immunity is initiated. In this work our aim was to analyze the surface charge of cells activated by carrageenan (CAR) and lipopolysaccharide (LPS) through light and electron microscopy approaches as well as the release of inflammatory mediators in vitro. The ultrastuctural analysis and the light microscopy data showed that in vivo administration of CAR represents a potent inflammatory stimulation for macrophages leading to a high degree of spreading, an increase in their size, in the number of the intracellular vacuoles and membrane projections as compared to the macrophages collected from untreated animals as well as mice submitted to LPS. Our data demonstrated that CAR stimulated-macrophages displayed a remarkable increase in nitric oxide production and PGE2 release as compared to the cells collected from non-stimulated and stimulated mice with LPS in vivo. On the other hand, non-stimulated macrophages as well as macrophages stimulated by LPS produce almost the same quantities of TNF-alpha, while in vivo stimulation by CAR leads to a 30-40% increase of cytokine release in vitro compared to the other groups. In conclusion, our morphological and biochemical data clearly showed that in vivo stimulation with CAR induces a potent inflammatory response in macrophages representing an interesting model to analyze inflammatory responses. PMID- 15342963 TI - Expression profile of mouse BWF1, a protein with a BEACH domain, WD40 domain and FYVE domain. AB - We isolated a mouse cDNA encoding a protein that contains a BEACH domain, 5 WD40 repeats and a FYVE domain, which we designated as BWF1. The mRNA is approximately 10 kb in size and encodes a protein consisting of 3508 amino acids with a predicted molecular weight of 385 kDa. BWF1 has 45% homology with the Drosophila protein, blue cheese (BCHS). The BWF1 gene consists of 67 exons, which span 270 kb of genomic sequence, and has been mapped to mouse chromosome 5. Northern blot analysis revealed that it was strongly expressed in the liver, moderately in the kidney and testis, and weakly in the brain of adult mice. During the development of the mouse brain, BWF1 mRNA was abundant on embryonic day (E) 14-16; after birth, the level of BWF1 mRNA expression decreased markedly to reach the adult level at postnatal day 3. In situ hybridization analysis revealed that the expressed BWF1 mRNA was restricted to the marginal region both in E14 and E16 embryonic brain, but became diffuse after birth. Confocal microscopy studies of the epitope-tagged BWF1 protein showed that the protein was a cytoplasmic one. PMID- 15342964 TI - An ultrastructural and biochemical study of foot structure in "catch" smooth muscle cells of a clam. AB - The foot structure of molluscan (clam) catch muscle cells was studied from the structural and biochemical standpoints. In vertebrate cross striated muscle cells, foot structures are situated in the interspaces between T-tubules and sarcoplasmic reticula (SRs). By contrast, T-tubules were not observed in clam catch muscle cells, but foot structures were ultrastructurally identified in the interspaces between the SRs and cell membranes. We isolated the SR fraction from muscle cells which contained vesicles with SRs and cell membranes. Foot structures were also observed in the SR fraction by thin sectioning. The size and shape of the foot structure in both intact muscle cells and the SR fractions appeared to be slightly smaller than those of vertebrates. However, the molecular weight of the foot structures (foot proteins) as determined by SDS-PAGE (450 kD) was similar to ryanodine receptors (RyRs) which were reported previously in cross striated muscle cells from pecten and vertebrates. The protein showing the 450 kD band reacted to an anti-ryanodine receptor by Western blotting. These findings are discussed in comparison with previous studies of foot structures and RyRs of vertebrates and invertebrates. PMID- 15342965 TI - Systematic analysis of SNARE molecules in Arabidopsis: dissection of the post Golgi network in plant cells. AB - In all eucaryotic cells, specific vesicle fusion during vesicular transport is mediated by membrane-associated proteins called SNAREs (soluble N-ethyl-maleimide sensitive factor attachment protein receptors). Sequence analysis identified a total of 54 SNARE genes (18 Qa-SNAREs/Syntaxins, 11 Qb-SNAREs, 8 Qc-SNAREs, 14 R SNAREs/VAMPs and 3 SNAP-25) in the Arabidopsis genome. Almost all of them were ubiquitously expressed through out all tissues examined. A series of transient expression assays using green fluorescent protein (GFP) fused proteins revealed that most of the SNARE proteins were located on specific intracellular compartments: 6 in the endoplasmic reticulum, 9 in the Golgi apparatus, 4 in the trans-Golgi network (TGN), 2 in endosomes, 17 on the plasma membrane, 7 in both the prevacuolar compartment (PVC) and vacuoles, 2 in TGN/PVC/vacuoles, and 1 in TGN/PVC/plasma membrane. Some SNARE proteins showed multiple localization patterns in two or more different organelles, suggesting that these SNAREs shuttle between the organelles. Furthermore, the SYP41/SYP61-residing compartment, which was defined as the TGN, was not always located along with the Golgi apparatus, suggesting that this compartment is an independent organelle distinct from the Golgi apparatus. We propose possible combinations of SNARE proteins on all subcellular compartments, and suggest the complexity of the post Golgi membrane traffic in higher plant cells. PMID- 15342966 TI - Induction chemotherapy with docetaxel, cisplatin and 5-fluorouracil for tongue cancer. AB - Induction chemotherapy consisting of one cycle of docetaxel, cisplatin and 5 fluorouracil (5-FU) was evaluated for its primary effects on squamous cell carcinoma of the tongue. The patients were five men and one woman, 50 to 67 (mean 57.8) years of age. Clinical staging of the tongue cancers showed two patients with stage III and four with stage IV disease. All patients underwent one cycle of intravenous chemotherapy with docetaxel (60 mg/m2, on day 1), cisplatin (10 mg/m2, from days 1 to 5) and 5-FU (500 mg/body, from days 1 to 5) before surgical operation. The overall response rate was 83.3% (five PRs and one NC), with two cases with grade II A in the Ohboshi & Shimosato classification and four with grade II B. The main side effect was severe neutropenia, which was effectively managed with granulocyte colony-stimulating factor (G-CSF). In conclusion, induction chemotherapy with docetaxel, cisplatin and 5-FU for tongue cancer was tolerated well by all the patients and showed an excellent response rate in spite of only one cycle of administration. PMID- 15342967 TI - The role of human papilloma virus in the molecular biology of cervical carcinogenesis. AB - Research exploring the E6-p53 and E7-pRb model has resulted in the identity of the viral gene's actions on numerous cellular proteins and processes normally involved in cell growth and proliferation. Specially, several findings have established the various ways by which the HPV-infected cell may escape controls governing cell growth and proliferation, including the fidelity of the host cell's genome and apoptosis. A large body of knowledge already generated in this area supports the view that high-risk HPV types have the ability to transform cells into a malignant phenotype. Such ability, however, is not sufficient to actually and inevitably produce cervical carcinoma, as indicated by the frequent spontaneous clearance of HPV infection and the long delay between the onset of persistent infection and emergence of the malignancy. Delay in the participation of cofactors has been suggested as explanation in this regard. However, it remains unclear how and when cofactors or factors that are innate in the HPV infected cells launch the host cells into an irreversible progression to carcinoma. PMID- 15342968 TI - PDGF-alpha receptor expression following hypoxic-ischemic injury in the neonatal rat brain. AB - Hypoxia-ischemia (HI) causes injury to oligodendrocytes (OLs), cells which create the myelin sheath in the developing brain. OLs pass successively through progenitor and immature stages during differentiation into mature OLs. Only the OLs in the progenitors stage can express the platelet-derived growth factor-a receptor (PDGF-R(alpha)) so that its expression is one of the cellular markers of OL progenitors. Activation of PDGF-R(alpha) results in OL proliferation, but not OL differentiation. To study the response of OL progenitors after neonatal HI brain injury, we investigated the expression of PDGF-R(alpha) in a neonatal rat stroke model (combination of left common carotid artery ligation and exposure to 8% O2 for 2 h). In the injured cerebral cortex, PDGF-R(alpha) mRNA levels increased significantly (p<0.01) with a peak at 0.5 h after HI insult, and returned to baseline levels within 48 h post-injury. PDGF-R(alpha) protein levels increased significantly at 72-96 h (p<0.05) and then returned to basal levels. Immunohistochemistry showed clear staining of PDGF-R(alpha) only in the injured cerebral cortex at 72 h after HI insult. In contrast, no staining was observed in the cortex of sham-operated controls. These results indicate that the expression of PDGF-R(alpha) increases rapidly and transiently only in the injured cerebral cortex after HI insult and may play a protective role through modulating the glial differentiation under the condition of cellular damage in the developing brain. PMID- 15342969 TI - Patient and family fact sheet. Traumatic brain injury. PMID- 15342970 TI - Acquired methemoglobinemia: a retrospective series of 138 cases at 2 teaching hospitals. AB - Methemoglobin is a form of hemoglobin that does not bind oxygen. When its concentration is elevated in red blood cells, functional anemia and tissue hypoxia may occur. We performed a retrospective case series to describe the cases of acquired methemoglobinemia (methemoglobin level >2%) detected and the clinical circumstances under which they occurred at 2 tertiary care hospitals and affiliated outpatient clinics over 28 months. We surveyed co-oximetry laboratory data to identify patients with methemoglobinemia. We reviewed these patients' medical records to extract the clinical information and context. One hundred thirty-eight cases of acquired methemoglobinemia were detected over the 28 months. There was no gender predisposition, and the condition occurred over a wide range of ages (patients aged 4 days to 86 years). Cases occurred in many areas of the hospital, including outpatient clinics. One fatality and 3 near fatalities were directly attributable to methemoglobinemia. Dapsone was the most common etiology of acquired methemoglobinemia, accounting for 42% of all cases. The mean peak methemoglobin level among these individuals was 7.6%. In 5 of the patients with the most severely elevated levels, 20% benzocaine spray (Hurricaine Topical Anesthetic spray, Beutlich Pharmaceuticals, Waukegan, IL) was the etiology, associated with a mean peak methemoglobin level of 43.8%. Eleven pediatric patients developed methemoglobinemia either from exogenous exposure, such as drugs, or due to serious illness, such as gastrointestinal infections with dehydration. Almost all (94%) patients with methemoglobinemia were anemic. Drugs that cause acquired methemoglobinemia are ubiquitous in both the hospital and the outpatient setting. Acquired methemoglobinemia is a treatable condition that causes significant morbidity and even mortality. We hope that a heightened awareness of methemoglobinemia will result in improved recognition and treatment. Primary prevention efforts have the potential to reduce the morbidity and mortality associated with this condition. PMID- 15342971 TI - Serial lactate determinations for prediction of outcome after cardiac arrest. AB - We investigated the relationship between lactate clearance and outcome in patients surviving the first 48 hours after cardiac arrest. We conducted the study in the emergency department of an urban tertiary care hospital. We analyzed the data for all 48-hour survivors after successful resuscitation from cardiac arrest during a 10-year period. Serial lactate measurements, demographic data, and key cardiac arrest data were correlated to survival and best neurologic outcome within 6 months after cardiac arrest. Parameters showing significant results in univariate analysis were tested for significance in a logistic regression model. Of 1502 screened patients, 394 were analyzed. Survivors (n = 194, 49%) had lower lactate levels on admission (median, 7.8 [interquartile range, 5.4-10.8] vs 9 [6.6-11.9] mmol/L), after 24 hours (1.4 [1-2.5] vs 1.7 [1.1 3] mmol/L), and after 48 hours (1.2 [0.9-1.6] vs 1.5 [1.1-2.3] mmol/L). Patients with favorable neurologic outcome (n = 186, 47%) showed lower levels on admission (7.6 [5.4-10.3] vs 9.2 [6.7-12.1] mmol/L) and after 48 hours (1.2 [0.9-1.6] vs 1.5 [1-2.2] mmol/L). In multivariate analysis, lactate levels at 48 hours were an independent predictor for mortality (odds ratio [OR]: 1.49 increase per mmol/L, 95% confidence interval [CI]: 1.17-1.89) and unfavorable neurologic outcome (OR: 1.28 increase per mmol/L, 95% CI: 1.08-1.51). Lactate levels higher than 2 mmol/L after 48 hours predicted mortality with a specificity of 86% and poor neurologic outcome with a specificity of 87%. Sensitivity for both end points was 31%. Lactate at 48 hours after cardiac arrest is an independent predictor of mortality and unfavorable neurologic outcome. Persisting hyperlactatemia over 48 hours predicts a poor prognosis. PMID- 15342972 TI - Neurologic manifestations in primary Sjogren syndrome: a study of 82 patients. AB - Neurologic involvement occurs in approximately 20% of patients with primary Sjogren syndrome (SS). However, the diagnosis of SS with neurologic involvement is sometimes difficult, and central nervous system (CNS) manifestations have been described rarely. We conducted the current study to describe the clinical and laboratory features of SS patients with neurologic manifestations and to report their clinical outcome. We retrospectively studied 82 patients (65 women and 17 men) with neurologic manifestations associated with primary SS, as defined by the 2002 American-European criteria. The mean age at neurologic onset was 53 years. Neurologic involvement frequently preceded the diagnosis of SS (81% of patients). Fifty-six patients had CNS disorders, which were mostly focal or multifocal. Twenty-nine patients had spinal cord involvement (acute myelopathy [n = 12], chronic myelopathy [n = 16], or motor neuron disease [n = 1]). Thirty-three patients had brain involvement and 13 patients had optic neuropathy. The disease mimicked relapsing-remitting multiple sclerosis (MS) in 10 patients and primary progressive MS in 13 patients. We also recorded diffuse CNS symptoms: some of the patients presented seizures (n = 7), cognitive dysfunction (n = 9), and encephalopathy (n = 2). Fifty-one patients had peripheral nervous system involvement (PNS). Symmetric axonal sensorimotor polyneuropathy with a predominance of sensory symptoms or pure sensory neuropathy occurred most frequently (n = 28), followed by cranial nerve involvement affecting trigeminal, facial, or cochlear nerves (n = 16). Multiple mononeuropathy (n = 7), myositis (n = 2), and polyradiculoneuropathy (n = 1) were also observed. Thirty percent of patients (all with CNS involvement) had oligoclonal bands. Visual evoked potentials were abnormal in 61% of the patients tested. Fifty-eight patients had magnetic resonance imaging (MRI) of the brain. Of these, 70% presented white matter lesions and 40% met the radiologic criteria for MS. Thirty-nine patients had a spinal cord MRI. Abnormalities were observed only in patients with spinal cord involvement. Among the 29 patients with myelopathy, 75% had T2-weighted hyperintensities. Patients with PNS manifestations had frequent extraglandular complications of SS. Anti-Ro/SSA or anti-La/SSB antibodies were detected in 21% of patients at the diagnosis of SS and in 43% of patients during the follow-up (mean follow-up, 10 yr). Biologic abnormalities were more frequently observed in patients with PNS involvement than in those with CNS involvement (p < 0.01). Fifty-two percent of patients had severe disability, and were more likely to have CNS involvement than PNS involvement (p < 0.001). Treatment by cyclophosphamide allowed a partial recovery or stabilization in patients with myelopathy (92%) or multiple mononeuropathy (100%). The current study underlines the diversity of neurologic complications of SS. The frequency of neurologic manifestations revealing SS and of negative biologic features, especially in the event of CNS involvement, could explain why SS is frequently misdiagnosed. Screening for SS should be systematically performed in cases of acute or chronic myelopathy, axonal sensorimotor neuropathy, or cranial nerve involvement. The outcome is frequently severe, especially in patients with CNS involvement. Our study also underlines the efficacy of cyclophosphamide in myelopathy and multiple neuropathy occurring during SS. PMID- 15342973 TI - Acute renal embolism. Forty-four cases of renal infarction in patients with atrial fibrillation. AB - Acute renal embolus is rarely reported in the medical literature; thus, accurate data regarding presentation, laboratory tests, diagnostic techniques, and treatment are lacking. To better define this condition, we examined the medical records of all patients admitted to Kaplan Medical Center and Sheba Medical Center in central Israel from 1984 to 2002 who had a diagnosis of renal infarction and atrial fibrillation. We noted demographic, clinical, and laboratory parameters; method of diagnosis; treatment received; and patient outcome. We identified 44 cases of renal embolus: 23 females and 21 males, with an average age of 69.5 +/- 12.6 years. Thirty (68%) patients had abdominal pain, and 6 (14%) had a previous embolic event. Nine patients were being treated with warfarin on admission, 6 (66%) of whom had an international normalized ratio (INR) < 1.8. Hematuria was present in 21/39 (54%), and 41 (93%) patients had a serum lactate dehydrogenase (LDH) level > 400 U/dL. The mean LDH was 1100 +/- 985 U/dL. Diagnostic techniques included renal isotope scan, which was abnormal in 36/37 cases (97%); contrast-enhanced computed tomography (CT) scan, which was diagnostic in 12/15 cases (80%); and ultrasound, which was positive in only 3/27 cases (11%). Angiography was positive in 10/10 cases (100%). Twenty-three (61%) of 38 patients had normal renal function on follow-up. The 30-day mortality was 11.4%. Renal embolus was diagnosed mainly in patients aged more than 60 years, some of whom had a previous embolic event. Most of those receiving anticoagulant therapy had a subtherapeutic INR. Abdominal pain was common, as well as hematuria and an elevated LDH. These patients are at risk of subsequent embolic events to other organs. The most sensitive diagnostic technique in this population is a renal isotope scan, but contrast-enhanced CT scan requires further assessment. PMID- 15342974 TI - A case series and focused review of nocardiosis: clinical and microbiologic aspects. AB - Nocardia species are ubiquitous soil organisms that often infect patients with underlying immune compromise, pulmonary disease, or a history of surgery or trauma. We report 5 cases of nocardiosis representing various aspects of this "great imitator": 1) pneumonia in the setting of underlying malignancy, 2) chronic pneumonia with drug-resistant organism, 3) bacteremia and empyema with chronic hematologic malignancy, 4) primary cutaneous disease, and 5) sternal wound infection. We present a summary of the English literature from 1966 to 2003 with a focus on the teaching points of each of our 5 cases as well as the background epidemiology and microbiology of the Nocardia genus. Isolation of the organism may be achieved with routine media but longer incubation times may be necessary, delaying diagnosis and appropriate therapy. Treatment with a sulfa containing regimen is standard of care, but resistance testing is warranted given emerging drug resistance, high rates of discontinuation due to adverse reactions, and the potential for nephrotoxicity in transplant recipients on cyclosporine. PMID- 15342975 TI - A unique immunofluorescence method promotes accurate diagnosis in MYH9 disorders: a case report. AB - The identification of a mutation in the MYH9 gene in hereditary macrothrombocytopenia has established a distinct entity proposed as "MYH9 disorders," which previously have often been misdiagnosed as chronic immune thrombocytopenic purpura. The authors describe clinical and laboratory characterization of a family with the disorder demonstrating giant platelets, thrombocytopenia, and leukocyte inclusion bodies. The authors emphasize the efficacy of a unique immunofluorescence method for the nonmuscle myosin heavy chain A in the diagnosis, because it is more sensitive than May-Grunwald-Giemsa staining and more practical than electron microscopy or direct sequencing. MYH9 disorders may be much more common than previously realized if accurately diagnosed. PMID- 15342977 TI - Prenatal diagnosis in Li-Fraumeni syndrome. AB - PURPOSE: The hallmark of Li-Fraumeni syndrome (LFS), a familial cancer syndrome, is constitutional TP53 mutation. The authors addressed the complex question of predictive prenatal genetic testing for cancer risk associated with inheritance of TP53 mutation. METHODS: A classic LFS family including the proband (a 20-month old boy with rhabdomyosarcoma), his 36-year-old father with osteosarcoma, and his 40-year-old paternal aunt with bilateral breast cancer were identified as carriers of a TP53 germline mutation, a novel 1 base pair deletion in exon 5. A few years later, the mother became pregnant twice, and the parents requested prenatal diagnosis on each occasion. Genetic counseling, psychological evaluation, and support were provided by a multidisciplinary team including a pediatric oncologist, a geneticist, a psychosocial worker, a prenatal care provider, and an ethical representative. After providing overall information on LFS, including the high risk of developing secondary multiple neoplasms in LFS survivors, the committee approved prenatal diagnosis at the request of the family. RESULTS: In the two pregnancies, the two fetuses were found to be carriers of the same mutation. Nine years from diagnosis of the first tumor, the proband, and a month later his father, developed second tumors, multifocal osteosarcoma and leiomyosarcoma, respectively. CONCLUSIONS: Children with primary tumors belonging to LFS should be considered for screening for germline mutations and genetic counseling by a multidisciplinary team. Whether family members are found to be positive or negative as carriers, such measures may provide, by reducing uncertainty, psychological benefit to high-risk families. PMID- 15342978 TI - Predictive genetic testing for childhood cancer: taking the road less traveled by. PMID- 15342979 TI - Ataxia and secretory diarrhea: two unusual paraneoplastic syndromes occurring concurrently in the same patient with ganglioneuroblastoma. AB - The presence of rare paraneoplastic syndromes, the opsoclonus-myoclonus-ataxia syndrome (OMA), presumably caused by antineuronal antibody production, and diarrhea, caused by vasoactive intestinal peptide (VIP) secreted by neuroblastoma, may strongly signal the presence of neuroblastoma. The authors describe a child who presented with both syndromes concurrently; this has never been described previously in the same patient. However, diagnosis of neuroblastoma was delayed by a workup focused on the prolonged diarrhea rather than the ataxia. The diarrhea resolved after tumor resection, whereas OMA required further therapy. Increased awareness of VIP-secretory diarrhea, especially in an ataxic child, might contribute to an earlier diagnosis of neuroblastoma. PMID- 15342980 TI - Paraneoplastic syndromes in neuroblastoma. PMID- 15342981 TI - Intensive chemotherapy without radiotherapy gives more than 85% event-free survival for non-Hodgkin lymphoma without central nervous involvement: a 6-year population-based study from the nordic society of pediatric hematology and oncology. AB - BACKGROUND: The prognosis in childhood non-Hodgkin lymphoma (NHL) has improved dramatically during recent decades. The authors report the results from a 6-year population-based study of clinical characteristics and treatment results of NHL from the five Nordic countries. METHODS: All children younger than 15 years of age at diagnosis with NHL diagnosed from 1995 to 2000 were stratified and treated according to immunophenotypic classification and stage of disease. RESULTS: A total of 230 patients were diagnosed with primary NHL, which gives an annual incidence of 0.9/100.000 children, with a median age of 8 years. Seven percent of the children were below 3 years of age at diagnosis. The male/female ratio was 2.3 and was unrelated to age. Patients with pre-B and T-cell NHL constituted 33%, B-cell NHL 53%, and anaplastic large cell lymphoma (ALCL) 14%. According to Murphy's classification, 14% had stage 1, 17% stage 2, 50% stage 3, and 19% stage 4 disease, 12 of whom (28%) had central nervous involvement (CNS) at diagnosis. By January 1, 2003, four children had died during induction, three children died in remission (2, 6, and 26 months from diagnosis), and 24 children experienced a relapse. At 5 years, the probability of event-free survival (p-EFS) was 86+/-2% for all children. The 5-year p-EFS values for stages 1 through 4 were 94%, 97%, 83%, and 79%, respectively. The 5-year p-EFS values were 91% for B-cell, 87% for pre-B, 81% for ALCL, and 79% for T-cell NHL. The 12 patients with CNS involvement at diagnosis had a significantly poorer outcome than stage 4 patients with CNS involvement (p-EFS = 50% vs. 90%, P < 0.01). The 218 patients without CNS disease at diagnosis had a 5-year p-EFS of 88%. CONCLUSIONS: With modern intensive chemotherapy, more than 85% of NHL patients will achieve long-lasting first remission. In the future, preventing death during induction and remission and improving therapy for patients with CNS disease would have a major impact on the overall p-EFS. PMID- 15342982 TI - An assessment of published pediatric dosage guidelines for enoxaparin: a retrospective review. AB - OBJECTIVE: To evaluate the ability of published dosage guidelines for enoxaparin to achieve therapeutic anticoagulation and to determine whether the routine monitoring of anti-Xa levels is still necessary at a tertiary care pediatric institution. METHODS: Consecutive charts and laboratory records were reviewed for all patients receiving treatment doses of enoxaparin for thrombosis in the authors institution over a 4-year period (1998-2002). RESULTS: Sixty-six percent (25/38) of the anti-Xa levels were within the recommended therapeutic range (0.5 1.0 [+/- 10%] U/mL) after two doses. The success rates of achieving therapeutic levels were 1/6, 2/3, 6/9, 10/11, and 6/9, for patients 2 months or younger, more than 2 months to 1 year, more than 1 year to 6 years, more than 6 years to 12 years, and more than 12 years of age, respectively. Patients with cardiac or renal disease were more likely to achieve high anti-Xa levels. Thirty-seven percent of patients reported adverse effects. The most common effects were injection site-related bruising and minor bleeding. One patient experienced a major bleed that was not life-threatening. CONCLUSIONS: Most patients achieved therapeutic anticoagulation when dosed according to the published guidelines. Children with cardiac conditions or renal insufficiency or those younger than 2 months were more likely to require dosage adjustments to achieve the therapeutic range. Routine monitoring of anti-Xa levels is still necessary in these patient populations, particularly when the early establishment of therapeutic anticoagulation may be critical. Enoxaparin appears to be well tolerated in the authors' patient population. PMID- 15342983 TI - Increased IL-16 levels in hemophagocytic lymphohistiocytosis. AB - Serum levels of interleukin-16 (IL-16) were measured to investigate its role in the pathophysiology of hemophagocytic lymphohistiocytosis (HLH). Serum IL-16 levels in patients with acute HLH were significantly higher than those in healthy controls and patients with infectious mononucleosis. They returned to normal levels in the convalescent phase of the disease. In contrast to serum interferon gamma (IFN-gamma) levels, serum IL-16 levels showed a gradual decrease over the course of the disease. Serum IL-16 levels showed a significant positive correlation with serum levels of soluble IL-2 receptor, IFN-gamma, and interleukin-18, body temperature, and serum lactic dehydrogenase (LDH) levels. An increase in IL-16 mRNA expression was detected in the liver of an HLH patient. These results suggest that IL-16 plays an important role in the pathophysiology of HLH by TH1 cell recruitment and activation at organs with inflammation. PMID- 15342984 TI - Validity of the child health questionnaire for use in children with sickle cell disease. AB - OBJECTIVE: To provide accurate data on health-related quality of life (HRQL), there must be a valid tool to measure this outcome. The objective of this study was to determine the validity of the Child Health Questionnaire (CHQ) as a measure of HRQL in sickle cell disease (SCD) by examining the relationship between HRQL and disease severity. METHODS: This was a cross-sectional study of children conducted at two urban, hospital-based clinics. The study participants were children with SCD ages 5 to 18 years who presented for a routine visit to the comprehensive SCD clinic. The main outcome was HRQL, as measured by the CHQ Parent Form 28 (PF28). A t test was used to compare HRQL between those with mild and severe disease. RESULTS: Parents/caretakers of 95 children completed the CHQ PF28. Children with mild SCD had a significantly better HRQL, as evidenced by a higher mean physical summary score (39.1), than those with severe disease (28.0) (difference=11.1, 95% confidence interval 5.03-18.11). There was no significant difference in psychosocial summary scores between groups. CONCLUSIONS: The CHQ is a valid tool to assess HRQL in children with SCD and could serve as an important adjunct to determine the effect of SCD on the lives of children. PMID- 15342985 TI - KL-6: marker for pulmonary involvement in Langerhans cell histiocytosis in infants. AB - The purpose of this study was to investigate serum KL-6 levels in an infant with Langerhans cell histiocytosis (LCH) and pulmonary involvement. The histologic diagnosis of LCH was established by skin biopsy. Imaging of the chest confirmed marked interstitial shadowing throughout both lung fields. Acutely, serum KL-6 was elevated to 9,400 U/mL. Following induction chemotherapy, clinical manifestations of LCH improved and the levels of serum KL-6 returned to within normal limits. During the maintenance therapy phase, there was a resurgence of the LCH, but without involvement of the lungs, and the levels of KL-6 remained normal. The authors conclude that KL-6 may be a useful marker for pulmonary involvement in infants with LCH. PMID- 15342986 TI - Bullous herpes zoster in a child with leukemia: case report and review of the literature. AB - Blistering disorders in childhood are usually difficult to diagnose and pose complicated management dilemmas. The incidence of herpes zoster in children with malignancy and immunodeficiency is greatly increased compared to normal children of comparable age. Although herpes zoster is known to occur in children with malignancy, the bullous form of herpes zoster is rare; to the authors' knowledge, there was no previous report of this phenomenon in children in general and in children with cancer in particular. The authors describe a 3.5-year-old girl who was diagnosed with acute lymphoblastic leukemia; 7 months after presentation, during chemotherapy treatment, she developed the bullous form of herpes zoster on her right hand. The authors describe the method of diagnosis and provide a review of the literature concerning this rare phenomenon. Recognizing this entity and differentiating it from other bullomatous conditions enable the application of appropriate treatment, without unnecessary delay. PMID- 15342987 TI - Juvenile myelomonocytic leukemia presenting with features of hemophagocytic lymphohistiocytosis in association with neurofibromatosis and juvenile xanthogranulomas. AB - An association exists among neurofibromatosis 1 (NF1), juvenile xanthogranulomas (JXGs), and juvenile myelomonocytic leukemia (JMML). The authors describe a patient with the triple association of JXG, NF1, and JMML initially presenting with features of hemophagocytic lymphohistiocytosis (HLH). An 18-month old boy had multiple cutaneous and gastrointestinal JXG and NF1. At 3 years of age he developed anemia, thrombocytopenia, and hepatosplenomegaly. A bone marrow biopsy revealed features of HLH. Despite chemotherapy, he went on to develop JMML, which proved fatal. PMID- 15342988 TI - Topotecan disposition in an anephric child. AB - Although limited data are available about topotecan disposition in patients with renal insufficiency, nothing has been reported in anephric patients. The objective of this report is to characterize topotecan disposition in an anephric child with Wilms tumor, both on and off hemodialysis. The patient received topotecan and cyclophosphamide for four cycles; topotecan was administered daily for 5 days, with hemodialysis on the second and fourth day. Therapy was well tolerated, with grade 3 thrombocytopenia and grade 2 neutropenia noted after cycle four. The median topotecan lactone clearance was 15.5 L/h/m off hemodialysis and 18.7 L/h/m on hemodialysis. Topotecan clearance was minimally affected by hemodialysis and was similar to that observed in children without renal failure. PMID- 15342989 TI - Emergence and compartmentalization of fatal multi-drug-resistant cytomegalovirus infection in a patient with autosomal-recessive severe combined immune deficiency. AB - The authors describe a patient with autosomal-recessive severe combined immunodeficiency (SCID) with severe, multiorgan cytomegalovirus (CMV) disease. In the face of appropriate therapy, the patient developed a 100-fold gradient in viral load across the blood-brain barrier. Disseminated disease, including pneumonitis, contributed to a fatal outcome. Serial genotypic analyses revealed multiple UL97 and UL54 (DNA polymerase) mutations that conferred phenotypic resistance to all currently licensed systemic CMV antivirals. PMID- 15342990 TI - A case of juvenile myelomonocytic leukemia with concomitant cytomegalovirus infection. AB - Infantile cytomegalovirus (CMV)-associated disease and juvenile myelomonocytic leukemia (JMML) frequently present with similar clinical features, and thus the differential diagnosis is often difficult. An early and definite diagnosis of these disorders is required because their therapeutic approaches are very different. The authors describe a 2-month-old Japanese girl with JMML and CMV infection. The CMV antigen was detected by immunologic staining of leukocytes using the peroxidase-labeled monoclonal antibody HRP-C7. To assess clonality, the X-chromosome inactivation pattern was evaluated using polymerase chain reaction analysis of the human androgen receptor gene with or without predigestion of chromosomal DNA with HhaI or HpaII. The patient showed evidence of monoclonal origin of mononuclear cells at diagnosis. Although CMV infection mimicking JMML has previously been reported in two patients, to the authors' knowledge this is the first report describing a firm and definitive diagnosis of JMML based on the study of X-chromosome inactivation patterns. PMID- 15342991 TI - Usefulness of rh-G-CSF in early-onset severe neutropenia in neonatal lupus syndrome. AB - Neutropenia is a mild and transient manifestation of neonatal lupus syndrome (NLS) in the second or third month of life. The authors describe a newborn with an early-onset severe neutropenia due to anti-Ro/SSA. In the second day of life, neutropenia has been treated with recombinant human granulocyte colony stimulating factor (rh-G-CSF). This is the first case in which rh-G-CSF was used in NLS, and the authors studied the pharmacologic action of the drug in relationship to the pathophysiology of NLS. PMID- 15342994 TI - Induced seizures as psychiatric therapy: Ladislas Meduna's contributions in modern neuroscience. PMID- 15342995 TI - Learning from the history of neuroscience: dogma and patient interests: comment to Dr. Finks commentary. PMID- 15342996 TI - Bifrontal electroconvulsive therapy in the elderly: a 2-year retrospective. AB - OBJECTIVES: We sought to examine the clinical effect of bifrontal (BF) electroconvulsive therapy (ECT) in depressed patients aged 65 years and older. METHODS: A retrospective chart review of all patients who received BF ECT for a depressive disorder between January 2000 and December 2002 was made. RESULTS: Fourteen patients, with a mean age of 73.9 years, received BF ECT. Nine had unipolar and 5 had bipolar depression. Twelve patients (86%) responded unequivocally after a mean of 8.5 treatments. Five (35%) experienced cognitive side effects. Ninety-two percent of patients were discharged on lithium (0.6 mmol/L) with 86% also receiving antidepressants or antipsychotics. Only 1 patient relapsed, the remainder remaining well at follow-up after a mean of 18.7 months. CONCLUSION: BF ECT was found to be clinically effective and associated with cognitive side effects in elderly patients who were experiencing a depressive episode of either unipolar or bipolar origin. PMID- 15342997 TI - Pretreatment neurophysiologic function and ECT response in depression. AB - OBJECTIVES: Recent brain imaging studies have provided evidence that brain function assessed prior to treatment of depression may be associated with eventual treatment response. The present study tested the hypothesis that brain activity in midline apical quantitative EEG (QEEG) electrodes would be associated with therapeutic response to electroconvulsive therapy (ECT). METHODS: Ten treatment-refractory patients with unipolar or bipolar depression received a Hamilton Rating Scale for Depression (Ham-D) at baseline, during, and following ECT treatment. Resting, eyes-closed, 35-lead QEEG recordings were done 1 day before the initial ECT treatment. Data were analyzed using QEEG power and cordance. RESULTS: The mean of the theta-band pretreatment cordance from the central brain region was strongly associated with percentage decrease in Ham-D score over the course of treatment (r = 0.80, P = 0.005). QEEG cordance from other brain regions and power from all brain regions did not show an association with clinical improvement. CONCLUSIONS: Depressed subjects with higher pretreatment central cordance appear to be more likely to experience therapeutic benefits of ECT. The location of central electrodes over the cingulate cortex may indicate that pretreatment cingulate activity is associated with response to ECT. PMID- 15342998 TI - Comparison of clinical efficacy and side effects for bitemporal and bifrontal electrode placement in electroconvulsive therapy. AB - OBJECTIVES: Bifrontal (BF) placement of electrodes in electroconvulsive therapy (ECT) has become a popular alternative to bitemporal (BT) placement. This study compares the clinical efficacy, side effects, and rehospitalization rates of BT and BF electrode placement in a community hospital setting. METHODS: Charts from 76 patients receiving ECT treatments at Harborview Medical Center from 1994 to 2000 were reviewed to extract data on the characteristics of the course of ECT, clinical response, total headaches, narcotic and nonsteroidal anti-inflammatory drug doses, as well as documentation of confusion, disorientation, memory loss, and treatment emergent need for assistance with activities of daily living. RESULTS: The BT patients experienced more clinical improvement during their stay (a 7-point greater change in Psychiatric Symptom Assessment Scale score, P < 0.05) and were significantly less likely to be rehospitalized within a 1-year time frame (odds ratio = 4.9, P = <0.05), even after controlling for relevant covariates. Although the two patient groups had equal rates of headache and analgesic administration, the BT placement caused significantly more cognitive impairment. CONCLUSIONS: This study suggests that BT electrode placement offers better efficacy but modestly greater cognitive impairment than BF electrode placement. PMID- 15342999 TI - A study into predictors for the speed of response to electroconvulsive therapy. AB - The efficacy of electroconvulsive therapy (ECT) has been firmly established in the treatment of depression. However, prediction of the speed of response to ECT is an issue that needs to be further explored. This study aims to examine the presence of predictors for the speed of response. In a retrospective chart review using 57 patients suffering from major depression who received ECT, the relation of several patient and ECT variables with the speed of response was explored. Response was defined as a drop of at least 35% in Hamilton rating scale of depression (HRSD) score from baseline after 3 or 4 ECT sessions. Patients received ECT with an aged-based stimulus dosage in a clinical setting. Multiple regression analysis showed that high baseline HRSD score and high seizure energy index (SEI) were significantly and independently associated with a rapid response. In a regression model, baseline HRSD score and SEI can be used to predict the speed of response to ECT. Rapid responders to ECT achieved remission significantly more often than slow responders did. SEI can be modified by the clinician. This offers the possibility to optimize ECT treatment. PMID- 15343000 TI - The maximum-likelihood strategy for determining transcranial magnetic stimulation motor threshold, using parameter estimation by sequential testing is faster than conventional methods with similar precision. AB - BACKGROUND: The resting motor threshold (rMT) is the basic unit of transcranial magnetic stimulation (TMS) dosing. Traditional methods of determining rMT involve finding a threshold of either visible movement or electromyography (EMG) motor evoked potentials, commonly approached from above and below and then averaged. This time-consuming method typically uses many TMS pulses. Mathematical programs can efficiently determine a threshold by calculating the next intensity needed based on the prior results. Within our group of experienced TMS researchers, we sought to perform an illustrative study to compare one of these programs, the Maximum-Likelihood Strategy using Parameter Estimation by Sequential Testing (MLS PEST) approach, to a modification of the traditional International Federation of Clinical Neurophysiology (IFCN) method for determining rMT in terms of the time and pulses required and the rMT value. METHODS: One subject participated in the study. Five researchers determined the same subject's rMT on 4 separate days twice using EMG and twice using visible movement. On each visit, researchers used both the MLS-PEST and the IFCN methods, in alternating order. RESULTS: The MLS PEST approach was significantly faster and used fewer pulses to estimate rMT. For EMG-determined rMT, MLS-PEST and IFCN derived similar rMT, whereas for visible movement MLS-PEST rMT was higher than for IFCN. CONCLUSIONS: The MLS-PEST algorithm is a promising alternative to traditional, time-consuming methods for determining rMT. Because the EMG-PEST method is totally automated, it may prove useful in studies using rMT as a quickly changing variable, as well as in large scale clinical trials. Further work with PEST is warranted. PMID- 15343001 TI - Electroconvulsive therapy in Scottish clinical practice: a national audit of demographics, standards, and outcome. AB - OBJECTIVES: Many of the commonly expressed concerns about the effectiveness of electroconvulsive therapy (ECT) have been addressed by scientific studies. However, this has done little to reassure service users and the public at large about its use in routine clinical practice. This 3-year study between 1997 and 1999 consisted of a series of audit cycles to systematically answer questions about ECT demographics and outcome across Scotland. METHODS: Audit standards were agreed and each ECT clinic was visited at least twice to assess facilities and adherence to protocol as set down by the Royal College of Psychiatrists, UK. Demographics and outcome after ECT were measured during 2 9-month sample periods. Adequacy of teaching and supervision was assessed directly and by questionnaire. RESULTS: Facilities and equipment at ECT centers were up to date and generally of a high standard. ECT was given at a rate of 142 treatments per 100,000, mainly to white adult patients suffering from a depressive disorder. The ratio of females to males was approximately 2 to 1, and ECT was not given disproportionately to the elderly. 76% of patients were of informal status and 82% gave informed consent, with the remaining 18% receiving treatment under the safeguards of the Mental Health (Scotland) Act 1984. There was a definite clinical improvement with treatment, defined as at least a 50% fall in the Montgomery Asberg Depression Scale, in 71% of those treated for depressive illness. 65% of those treated for a non -depressive psychosis were rated as at least 'much improved' on the Clinical Global Impression scale. CONCLUSIONS: ECT given in routine clinical settings across Scotland meets Royal College of Psychiatrists standards and is an effective treatment of the clear majority of patients. PMID- 15343002 TI - Electroconvulsive therapy, depression, and cognitive outcomes: an Australian audit. AB - OBJECTIVES: We sought to compare cognitive and other outcomes of 2 groups of mood disorder patients, those who received ECT and those who did not, from 2 private South Australian hospitals during a 12-month period. METHODS: Patients were assessed at admission and discharge from hospital on 2 validated instruments: the Health of the Nation Outcome Scales (HoNOS) and an abbreviated version of the Short-Form 36-item Health Status Questionnaire (SF-36). RESULTS: Patient scores on the majority of subscales of both the HoNOS and SF-36 indicated a positive response to either ECT or non-ECT. There was no significant difference in HoNOS depression score between the ECT and non-ECT groups upon discharge, despite the ECT group being significantly more depressed at admission. Cognitive functioning of those who received ECT did not change significantly from admission to discharge. However, they did not share the significant improvement in cognitive functioning of those depressed persons who had non-ECT treatment. CONCLUSIONS: ECT treatment of depression was not associated with a deterioration of cognitive functioning. PMID- 15343003 TI - One-year outcome of elderly inpatients with major depressive disorder treated with ECT and antidepressants. AB - We report the acute response and outcome in 1-year follow-up of 51 elderly depressive inpatients with major depressive disorder treated with electroconvulsive therapy (ECT) (n = 30) and/or antidepressant therapy (n = 21). The patients were assessed at admission, at discharge, and after 1 year according to the Montgomery and Asberg Depression Rating Scale, the Beck Depression Inventory, and the Clinical Global Impression Scale. The acute response was good. Montgomery and Asberg Depression Rating Scale total scores diminished significantly during index hospitalization within both groups (from 31.6 +/- 8.5, to 8.1 +/- 6.0 in the ECT group and from 28.5 +/- 5.4 to 13.4 +/- 10.6 in the antidepressant group). The 1-year rehospitalization rate for the entire group, however, was 21 of 51 patients (41%), 13 of 30 patients (43%) in the ECT group, and 8 of 21 (38%) in the antidepressant group. Six of 13 patients in the ECT group and 1 of 8 patients in the antidepressant group were rehospitalized during the first month after discharge. The results suggest a good acute therapeutic response to both ECT and antidepressive therapy in elderly patients with MDD. The major finding in this study was the relatively high rehospitalization rate, which emphasizes the need for careful follow-up of the elderly patients who have recovered from a depressive episode. PMID- 15343004 TI - Implications of herbal alternative medicine for electroconvulsive therapy. AB - OBJECTIVES: To clarify the implications of herbal alternative medicine use during electroconvulsive therapy (ECT). BACKGROUND: A substantial proportion of patients with mental disorders report frequent use of herbal alternative medicines. Our current understanding of the biology of such remedies suggests that they may have implications for ECT practice. METHODS: We conducted electronic literature searches using Medline (via PubMed), Cochrane Database of Systematic Reviews, ACP Journal Club, PsychINFO, Database of Abstracts of Reviews of Effects, and Cochrane Central Register of Controlled Trials from their inception to April 2003. The search items were five selected herbal alternative medicines (Ginkgo biloba, ginseng, St. John's wort, valerian, kava-kava) in combination with the terms "drug interaction," "adverse effects," "side effects," "adverse drug reactions," "safety," and "toxicity." All data were included regardless of whether they were case reports, case series, clinical trials, or reviews. RESULTS: Our literature review revealed several potential effects of herbal alternative medicines upon ECT outcome. CONCLUSIONS: The growing use of herbal alternative medicine by patients with psychiatric illness may have implications for ECT practice. Our current knowledge is sparse and incomplete, however, indicating the need for more research. PMID- 15343005 TI - Asystole and bradycardia related to anesthetic induction during ECT: a case report. PMID- 15343006 TI - Tumor necrosis factor-alpha, depression, and ECT: toward a better understanding of the relationships. PMID- 15343007 TI - Ventilator-associated lung injury in patients without acute lung injury at the onset of mechanical ventilation. AB - OBJECTIVE: Although ventilation with small tidal volumes is recommended in patients with established acute lung injury, most others receive highly variable tidal volume aimed in part at normalizing arterial blood gas values. We tested the hypothesis that acute lung injury, which develops after the initiation of mechanical ventilation, is associated with known risk factors for ventilator induced lung injury such as ventilation with large tidal volume. DESIGN: Retrospective cohort study. SETTING: Four intensive care units in a tertiary referral center. PATIENTS: Patients who received invasive mechanical ventilation for > or = 48 hrs between January and December 2001. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The main outcome of interest, acute lung injury, was assessed by independent review of daily digital chest radiographs and arterial blood gases. Ventilator settings, hemodynamics, and acute lung injury risk factors were extracted from the Acute Physiology and Chronic Health Evaluation III database and the patients' medical records. Of 332 patients who did not have acute lung injury from the outset, 80 patients (24%) developed acute lung injury within the first 5 days of mechanical ventilation. When expressed per predicted body weight, women were ventilated with larger tidal volume than men (mean 11.4 vs. 10.4 mL/kg predicted body weight, p <.001) and tended to develop acute lung injury more often (29% vs. 20%, p =.068). In a multivariate analysis, the main risk factors associated with the development of acute lung injury were the use of large tidal volume (odds ratio 1.3 for each mL above 6 mL/kg predicted body weight, p <.001), transfusion of blood products (odds ratio, 3.0; p < 0.001), acidemia (pH < 7.35; odds ratio, 2.0; p =.032) and a history of restrictive lung disease (odds ratio, 3.6; p =.044). CONCLUSIONS: The association between the initial tidal volume and the development of acute lung injury suggests that ventilator-associated lung injury may be an important cause of this syndrome. Height and gender should be considered when setting up the ventilator. Strong consideration should be given to limiting large tidal volume, not only in patients with established acute lung injury but also in patients at risk for acute lung injury. PMID- 15343008 TI - Persistent microcirculatory alterations are associated with organ failure and death in patients with septic shock. AB - OBJECTIVE: To characterize the time course of microcirculatory alterations and their relation to outcome in patients with septic shock. DESIGN: Prospective, observational study. SETTING: Thirty-one-bed, medico-surgical intensive care unit in a university hospital. PATIENTS: Forty-nine patients with septic shock. INTERVENTIONS: The sublingual microcirculation was investigated with an orthogonal polarization spectral imaging device on the day of onset of septic shock (baseline) and each day until resolution of shock. MEASUREMENTS AND MAIN RESULTS: Five sequences of 20 secs each were recorded and analyzed off-line by a semiquantitative method. Data were analyzed with nonparametric tests and presented as median (25th-75th percentiles). Three patients died after the resolution of shock from unrelated causes and were excluded. Of the other 46 patients, 26 survived and 20 died: 13 due to unresolving shock and seven due to persistent multiple organ failure after resolution of shock. At the onset of shock, survivors and nonsurvivors had similar vascular density (5.6 [4.7-7.0] vs. 6.2 [5.4-7.0]/mL; p = nonsignificant) and percentage of perfused small vessels (65.0 [53.1-68.9] vs. 58.4 [47.5-69.1]%; p = nonsignificant). Small vessel perfusion improved over time in survivors (analysis of variance, p <.05 between survivors and nonsurvivors) but not in nonsurvivors. Despite similar hemodynamic and oxygenation profiles and use of vasopressors at the end of shock, patients dying after the resolution of shock in multiple organ failure had a lower percentage of perfused small vessels than survivors (57.4 [46.6-64.9] vs. 79.3 [67.2-83.2]%; p =.02). CONCLUSIONS: Microcirculatory alterations improve rapidly in septic shock survivors but not in patients dying with multiple organ failure, regardless of whether shock has resolved. PMID- 15343009 TI - Half the family members of intensive care unit patients do not want to share in the decision-making process: a study in 78 French intensive care units. AB - OBJECTIVE: To evaluate the opinions of intensive care unit staff and family members about family participation in decisions about patients in intensive care units in France, a country where the approach of physicians to patients and families has been described as paternalistic. DESIGN: Prospective multiple-center survey of intensive care unit staff and family members. SETTING: Seventy-eight intensive care units in university-affiliated hospitals in France. PATIENTS: We studied 357 consecutive patients hospitalized in the 78 intensive care units and included in the study starting on May 1, 2001, with five patients included per intensive care unit. INTERVENTIONS: We recorded opinions and experience about family participation in medical decision making. Comprehension, satisfaction, and Hospital Anxiety and Depression Scale scores were determined in family members. MEASUREMENTS AND MAIN RESULTS: Poor comprehension was noted in 35% of family members. Satisfaction was good but anxiety was noted in 73% and depression in 35% of family members. Among intensive care unit staff members, 91% of physicians and 83% of nonphysicians believed that participation in decision making should be offered to families; however, only 39% had actually involved family members in decisions. A desire to share in decision making was expressed by only 47% of family members. Only 15% of family members actually shared in decision making. Effectiveness of information influenced this desire. CONCLUSION: Intensive care unit staff should seek to determine how much autonomy families want. Staff members must strive to identify practical and psychological obstacles that may limit their ability to promote autonomy. Finally, they must develop interventions and attitudes capable of empowering families. PMID- 15343010 TI - A proactive approach to improve end-of-life care in a medical intensive care unit for patients with terminal dementia. AB - OBJECTIVES: To compare usual care with a proactive case-finding approach for critically ill patients with terminal dementia using an inpatient palliative care service. DESIGN: Prospective comparison to historical control. SETTING: Urban, university-affiliated hospital. PATIENTS: Total of 52 men and women with end stage dementia, 26 subjects in each control and intervention group. INTERVENTIONS: Proactive case-finding by the palliative care service was done to offer early assistance to the intensive care unit staff about the treatment of patients with terminal dementia. Results were compared with data obtained retrospectively. MEASUREMENTS AND MAIN RESULTS: Measurements included age, gender, Acute Physiology and Chronic Health Evaluation Score, Therapeutic Intervention Scoring System, mortality, intensive care unit and hospital lengths of stay, frequency, timing, and goals of do-not-resuscitate orders. The proactive, case-finding approach decreased hospital and medical intensive care unit length of stay. More important, a proactive palliative intervention decreased the time between identification of the poor prognosis and the establishment of do-not-resuscitate goals, decreased the time terminal demented patients remained in the intensive care unit, and reduced the use of nonbeneficial resources, thus reducing patient burden and the cost of care while having the potential to afford the patient and family increased comfort and psychoemotional support. CONCLUSIONS: Proactive interventions from a palliative care consultant within this subset of patients improved end-of-life care and decreased use of superfluous resources. PMID- 15343011 TI - Expiratory trigger setting in pressure support ventilation: from mathematical model to bedside. AB - OBJECTIVE: To evaluate the feasibility of relying on a mathematical model to adjust the optimal level of expiratory trigger, materialized by the ratio of inspiratory flow at the end of inspiratory effort (V'ti) and peak inspiratory flow (V'peak), or V'ti/V'peak, during pressure support, by comparing its predicted values with those measured in intubated patients. DESIGN: Prospective observational study. SETTING: Medical intensive care unit, university hospital. PATIENTS: There were 28 intubated patients undergoing pressure support. INTERVENTIONS: Pressure support as set by the clinician in charge. MEASUREMENTS AND MAIN RESULTS: A significant correlation was found between predicted and measured V'ti/V'peak ratios (r =.70; p <.001; mean +/- sd difference, -0.025 +/- 0.07; 95% confidence interval, -0.161 to 0.111). Overall, delayed cycling occurred in obstructive conditions, the delay increasing as obstructive disease was more severe. CONCLUSIONS: A significant correlation was observed between predicted values of V'ti/V'peak and those values measured in patients undergoing pressure support. These findings should stimulate further research into the possible applications of this mathematical model to optimize expiratory trigger setting. Furthermore, our findings suggest that expiratory trigger should be adjustable and provide a wider range of cutoff levels than that which is currently available. PMID- 15343012 TI - Effect of long-term and high-dose antithrombin supplementation on coagulation and fibrinolysis in patients with severe sepsis. AB - OBJECTIVE: Sepsis is frequently associated with coagulatory activation, which may contribute to deteriorated organ function. Antithrombin is one important endogenous coagulation inhibitor that is therapeutically applied during sepsis. This study investigates the effect of 14-day antithrombin application on coagulatory variables. DESIGN: Prospective study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: Forty patients with severe sepsis. INTERVENTIONS: Patients with severe sepsis were randomly assigned to receive either conventional intensive care treatment (n = 20, controls) or antithrombin substitution that aimed at a plasma antithrombin activity > or =120% during a long-term (14-day) study period (n = 20, antithrombin). To allow comparative analysis of laboratory variables over time, all patients who did not survive the 14-day-period (five controls and six antithrombin patients) were prospectively excluded from the final evaluation. Their data were included in an intent-to treat analysis. MEASUREMENTS AND MAIN RESULTS: Antithrombin supplementation normalized global coagulation tests and increased prothrombin activity as well as fibrinogen concentration, reflecting less coagulation factor consumption (percent change from baseline in prothrombin activity, p <.01 vs. controls at days 9, 11 14 of antithrombin vs. controls [unpaired Student's t-test]; fibrinogen concentration, p <.01 vs. controls at days 10, 11, 13, and 14 of antithrombin). Simultaneously, antithrombin reduced contact system activation as indicated by increasing prekallikrein activities over time (% change, p <.01 vs. controls at days 6, 9-14) and increased protein C activities when compared with controls (% change, p <.01 vs. controls at days 10-14). Most changes occurred from day 7 to day 14 of antithrombin supplementation. Antithrombin did not influence C1 esterase inhibitor, plasminogen, alpha2 antiplasmin, or platelet counts (p >.01). CONCLUSION: In this first study on long-term antithrombin therapy, antithrombin significantly reduced septic coagulatory response in patients with severe sepsis when given over 14 days. PMID- 15343013 TI - Myocardial infarction rate in acute pulmonary edema: noninvasive pressure support ventilation versus continuous positive airway pressure. AB - OBJECTIVE: Noninvasive pressure support ventilation (NIPSV) delivered by face mask has proved an effective treatment for patients with acute pulmonary edema. However, an increase in acute myocardial infarction rate has been reported with this ventilation modality. We investigated whether the use of NIPSV increases the incidence of acute myocardial infarction compared with continuous positive airway pressure (CPAP) in patients with acute pulmonary edema. DESIGN: Randomized, prospective, controlled study. SETTING: Emergency Department, Niguarda Hospital of Milano (Italy). PATIENTS: Forty-six patients affected by acute pulmonary edema. INTERVENTIONS: The patients received either NIPSV (24 patients) or CPAP (22 patients) through a face mask. MEASUREMENTS AND MAIN RESULTS: Cardiac enzymes (myoglobin, creatine kinase isoenzyme MB, and troponin I) were determined and electrocardiographic and physiologic measurements made over the subsequent 36 hrs. No significant differences were observed in the incidence of acute myocardial infarction in the CPAP group (13.6%) compared with the NIPSV group (8.3%). Both modalities of noninvasive ventilation improved ventilation and vital signs in patients with acute pulmonary edema. Two patients of the NIPSV group (8.3%) and one of the CPAP group (4.5%) required endotracheal intubation because vital signs and arterial blood gases worsened 1 hr after the start of noninvasive ventilation. No significant differences were found in in-hospital mortality rate. CONCLUSIONS: NIPSV proved to be equally effective in improving vital signs and ventilation without increasing acute myocardial infarction rate in patients with nonischemic acute pulmonary edema in comparison to CPAP alone. However, because the study lacked statistical power and excluded patients with acute coronary syndromes, caution is still advised when applying NIPSV to the latter subgroup of patients. PMID- 15343014 TI - Postoperative troponin-T predicts prolonged intensive care unit length of stay following cardiac surgery. AB - OBJECTIVE: To evaluate the use of postoperative cardiac troponin T (cTnT) for the prediction of prolonged intensive care unit length of stay following cardiac surgery. DESIGN: Prospective, single-center, observational cohort study of patients following cardiac surgical procedures. The enrollment period was from October through December 2000. Patients were enrolled on admission to the intensive care unit and followed until hospital discharge. SETTING: The cardiac surgical intensive care unit of the Massachusetts General Hospital. PATIENTS: A total of 222 consecutive patients were enrolled. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Perioperative clinical factors and serum concentrations of cTnT measured every 8 hrs after surgery were recorded. These clinical factors and the results of serum cTnT measurement were correlated with the need for prolonged intensive care unit length of stay (defined as >24 hrs). Univariable analysis identified factors predictive of prolonged intensive care unit length of stay. Stepwise logistic regression identified independent predictors of prolonged intensive care unit length of stay. Multiple linear regression was used to explore the direct relationship between cTnT concentrations at several postoperative time points and intensive care unit length of stay. At each time point assessed, cTnT concentrations from patients requiring a prolonged intensive care unit length of stay were significantly higher (all p <.001) than in those individuals with normal length of stay. In contrast, creatine kinase isoenzymes were not significantly different between patients with normal or prolonged intensive care unit length of stay. Multivariable analysis demonstrated that an immediate postoperative cTnT concentration > or =1.58 ng/mL was the strongest predictor of a prolonged intensive care unit length of stay (odds ratio, 5.6; 95% confidence interval, 2.9 10.8). Multiple linear regression analysis revealed that intensive care unit length of stay increased by 0.32 days with each incremental 1.0 ng/mL increase in cTnT measured at 18-24 hrs postprocedure. CONCLUSIONS: Elevated postoperative cTnT concentrations can prospectively identify patients requiring prolonged intensive care unit length of stay after cardiac surgery. PMID- 15343015 TI - Effects of L-NAME and inhaled nitric oxide on ventilator-induced lung injury in isolated, perfused rabbit lungs. AB - OBJECTIVE: To determine whether nitric oxide (NO) might modulate ventilator induced lung injury. DESIGN: Randomized prospective animal study. SETTING: Animal research laboratory in a university hospital. SUBJECTS: Isolated, perfused rabbit heart-lung preparation. INTERVENTIONS: Thirty-six isolated, perfused rabbit lungs were randomized into six groups (n = 6) and ventilated using pressure-controlled ventilation for two consecutive periods (T1 and T2). Peak alveolar pressure during pressure-controlled ventilation was 20 cm H2O at T1 and was subsequently (T2) either reduced to 15 cm H2O in the three low-pressure control groups (Cx) or increased to 25 cm H2O in the three high-pressure groups (Px). In the control and high-pressure groups, NO concentration was increased to approximately equal to 20 ppm (inhaled NO groups: CNO, PNO), reduced by NO synthase inhibition (L-NAME groups: CL-Name, PL-Name), or not manipulated (groups CE, PE). MEASUREMENTS AND MAIN RESULTS: Changes in ultrafiltration coefficients (deltaKf [vascular permeability index: g.min(-1).cm H2O(-1).100 g(-1)]), bronchoalveolar lavage fluid 8-isoprostane, and NOx (nitrate + nitrite) concentrations were the measures examined. Neither L-NAME nor inhaled NO altered lung permeability in the setting of low peak alveolar pressure (control groups). In contrast, L-NAME virtually abolished the change in permeability (deltaKf: PL-Name (0.10 +/- 0.03) vs. PNO [1.75 +/- 1.10] and PE [0.37 +/- 0.11; p <.05]) and the increase in bronchoalveolar lavage 8-isoprostane concentration induced by high-pressure ventilation. Although inhaled NO was associated with the largest change in permeability, no significant difference between the PE and PL-NAME groups was observed. The change in permeability (deltaKf) correlated with bronchoalveolar lavage NOx (r2 =.6; p <.001). CONCLUSIONS: L-NAME may attenuate ventilator induced microvascular leak and lipid peroxidation and NO may contribute to the development of ventilator-induced lung injury. Measurement of NO metabolites in the bronchoalveolar lavage may afford a means to monitor lung injury induced by mechanical stress. PMID- 15343016 TI - Influence of systemic inflammatory response syndrome on host resistance against bacterial infections. AB - OBJECTIVE: To determine the relationship between systemic inflammatory response syndrome (SIRS) and host innate immunities against bacterial infections. DESIGN: Controlled animal study. SETTING: University research laboratory. SUBJECTS: Male BALB/c mice, 8-10 wks of age. INTERVENTIONS: Morbidity and mortality rates of severe SIRS mice were compared with those of mild SIRS mice after infection with Enterococcus faecalis or methicillin-resistant Staphylococcus aureus (MRSA) or exposure to infectious complications induced by cecal ligation and puncture (CLP). In addition, a function of effector cells related to antibacterial innate immunities for these infections was analyzed in these two groups. Furthermore, SCIDbgMN mice (SCIDbg mice depleted of antibacterial effector cells) were reconstituted with effector cells from mild or severe SIRS mice and exposed to various infections. MEASUREMENTS AND MAIN RESULTS: Severe SIRS mice were greatly susceptible to E. faecalis, MRSA, and CLP-induced sepsis. On the other hand, as compared with normal mice, mild SIRS mice were resistant to these infections. All of SCIDbgMN mice inoculated with peritoneal macrophages (PMphi) from severe SIRS mice died after infection with E. faecalis or MRSA, whereas all SCIDbgMN mice inoculated with PMphi from mild SIRS mice survived after the same infection. SCIDbgMN mice that were inoculated with PMphi from normal mice and exposed to E. faecalis, MRSA, or CLP survived at rates of 50%, 50%, or 60%, respectively. PMphi from mild SIRS mice exhibited typical properties for classically activated macrophages (CAMphi), whereas those from severe SIRS mice exhibited typical properties for alternatively activated macrophages (AAMphi). CONCLUSIONS: Mphi associated host antibacterial innate immunities are greatly influenced by SIRS levels. CAMphi, effector cells for the antibacterial innate immunity against E. faecalis, MRSA, and CLP-induced sepsis, are induced in mild SIRS mice. AAMphi with no antibacterial capabilities are generated in mice with severe SIRS. Induction of CAMphi may protect severe SIRS patients against infections. PMID- 15343017 TI - Comparison between single-beat and multiple-beat methods for estimation of right ventricular contractility. AB - OBJECTIVE: It was investigated whether pharmacologically induced changes in right ventricular contractility can be detected by a so-called "single-beat" method that does not require preload reduction. DESIGN: Prospective animal research. SETTING: Laboratory at a large university medical center. SUBJECTS: Eight anesthetized pigs. INTERVENTIONS: End-systolic elastance values obtained by a recently proposed single-beat method (Eessb) were compared with those obtained using the reference multiple-beat method (Eesmb). MEASUREMENTS AND MAIN RESULTS: Administration of dobutamine increased Eesmb from 1.6 +/- 0.3 to 3.8 +/- 0.5 mm Hg/mL (p =.001), whereas there was only a trend toward an increase in Eessb from 1.5 +/- 0.2 to 1.7 +/- 0.4 mm Hg/mL. Esmolol decreased Eesmb from 1.7 +/- 0.3 to 1.1 +/- 0.2 mm Hg/mL (p =.006), whereas there was only a trend for a decrease in Eessb from 1.5 +/- 0.2 to 1.3 +/- 0.1. CONCLUSIONS: The present method using single-beat estimation to assess right ventricular contractility does not work as expected, since it failed to detect either increases or decreases in right ventricular contractility induced by pharmacologic interventions. PMID- 15343018 TI - Systemic and renal macro- and microcirculatory responses to arginine vasopressin in endotoxic rabbits. AB - OBJECTIVE: Arginine vasopressin is being used increasingly to treat vasodilatory hypotension, although little is known of its effects on regional perfusion. Arginine vasopressin hemodynamic effects in physiology are mainly mediated through the V1a receptor on blood vessels. To investigate this further, we studied the effect of arginine vasopressin on systemic and renal blood flow in anesthetized, ventilated rabbits given either intravenous saline or endotoxin, and the impact of blocking V1a receptors. DESIGN: Prospective, randomized, controlled study. SETTING: Animal research laboratory. SUBJECTS: Male White New Zealand rabbits. INTERVENTIONS: Measurement was made of mean arterial blood pressure, aortic and renal blood flow velocities (pulsed Doppler), and renal cortical and medullary flow (laser Doppler). MEASUREMENTS AND MAIN RESULTS: In a first series of animals, incremental intravenous boluses of arginine vasopressin ranging from 1 to 1000 ng were administered 90 mins postendotoxin or saline. In control rabbits (n = 9), increasing doses of arginine vasopressin elevated mean arterial blood pressure but reduced both aortic and renal blood flow velocity and renal cortical flow (p <.05). In endotoxic animals (n = 6), arginine vasopressin produced a similar increase in mean arterial blood pressure although aortic flow was maintained while renal blood flow velocity increased, mostly in its diastolic component (p <.05). Pretreatment with the V1a receptor antagonist in a second series of animals blunted all the effects observed in both control (n = 5) and endotoxic (n = 6) animals, suggesting that arginine vasopressin acted mainly through V1a subtype in this early phase of sepsis. CONCLUSIONS: Preservation of renal blood flow with arginine vasopressin during endotoxemia, in particular to the cortex, suggests it could be a promising agent for hemodynamic support during septic shock. PMID- 15343019 TI - Mild preseptic hypothermia is detrimental in rats. AB - OBJECTIVE: We evaluated the effects of mild hypothermia (32 degrees C), established before experimental intra-abdominal sepsis, on outcome, cytokine pattern, and muscle tissue oxygenation. DESIGN: Clinic modeling randomized laboratory trial. SETTING: University laboratory. SUBJECTS: Ninety-six male rats. INTERVENTIONS: In a group-sequential design, using 42 rats per group, we compared mild hypothermia with normothermia before peritonitis. Peritoneal inoculation with human stool bacteria was performed to simulate clinical trial conditions. Additionally, 12 rats underwent preoperative mild hypothermia without infection. MEASUREMENTS AND MAIN RESULTS: Primary end point was mortality at 120 hrs. Secondary end points were systemic cytokine concentrations, granulocyte counts, and muscle oxygen partial pressure. Survival rate was 40% (16 of 42) after preseptic hypothermia and 62% (26 of 42) after preseptic normothermia (p =.048). All hypothermic rats without infection survived. Interleukin-10 concentrations were 1843 +/- 96 pg/mL after preseptic hypothermia, 945 +/- 225 pg/mL with preseptic normothermia, and 520 +/- 121 pg/mL after hypothermia without infection (p<.001). Macrophage inflammatory protein-2 was comparable in the treatment groups. Interleukin-6 concentrations were 106 +/- 24 pg/mL after preseptic hypothermia and 276 +/- 76 pg/mL with preseptic normothermia (p<.05). Postinfection granulocyte count was 1.7 x 10(9)/L after hypothermia and 2.4 x 10(9)/L after normothermia (p =.2). After infection, muscle oxygen partial pressure was 47 +/- 10 mm Hg with preseptic hypothermia, 85 +/- 12 mm Hg in preseptic normothermia, and 49 +/- 9 mm Hg after hypothermia without infection (p =.7). CONCLUSIONS: In this rat model of intra-abdominal sepsis, mild preseptic hypothermia (32 degrees C) reduced survival, impaired granulocyte recruitment, and changed cytokine balance, suggesting immunosuppression. PMID- 15343020 TI - Importance of platelets and fibrinogen in neutrophil-endothelial cell interactions in septic shock. AB - OBJECTIVE: To examine the role of platelets, fibrin, and adhesion molecules in mediating neutrophil-endothelial cell interactions in septic shock. DESIGN: Controlled experiments using phase contrast microscopy to examine neutrophil, platelet, and endothelial cell interactions in flowing cell suspensions under simulated physiologic conditions. SETTING: University research laboratory. PATIENTS: Adult patients with septic shock and normal volunteers. INTERVENTIONS: Microslides were coated with human umbilical vein endothelial cells. Neutrophils were removed from control subjects and patients in septic shock and were perfused over endothelial cells at rates representing a range of physiologic shear stresses. In an attempt to examine the effects of fibrin deposition on neutrophil endothelial cell interactions, neutrophils, with and without platelets, were suspended in plasma and serum was removed from patients in septic shock. In addition, blocking monoclonal antibodies against the platelet receptor P-selectin and neutrophil receptor CD11b/CD18, and a platelet glycoprotein IIb/IIIa inhibitor, were incubated with cells suspended in plasma. Phase contrast video microscopy was used to count the number of neutrophils/mm adherent to endothelial cells during cessation of flow. Neutrophil rolling velocity was calculated as the time required for neutrophils to move across a 1-mm field (mm/sec). Leukoaggregation was defined as the number of neutrophils in aggregates (three or more nuclei) across a 1-mm field. MEASUREMENTS AND MAIN RESULTS: Normal neutrophils exposed to plasma from patients with septic shock demonstrated significant increases in aggregation and endothelial cell adherence with associated decreases in neutrophil rolling velocity. These changes were significantly enhanced in the presence of platelets and significantly attenuated in the presence of serum, which is fibrinogen depleted. Preincubation with antibodies to the surface receptors P-selectin, CD11b/CD18, and glycoprotein IIb/IIIa abrogated the changes in neutrophil aggregation, adhesion, and rolling velocity. CONCLUSIONS: These data suggest that platelets and fibrinogen play an important role in mediating neutrophil-endothelial cell adherence in septic shock. PMID- 15343021 TI - Effects of carbenoxolone on alveolar fluid clearance and lung inflammation in the rat. AB - OBJECTIVES: 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2), which requires oxidized nicotinamide adenine dinucleotide as a cofactor, metabolizes endogenous glucocorticoids. Since 11beta-HSD2 has been detected in lung epithelial cells, we examined whether carbenoxolone, a potent inhibitor of 11beta HSD, would enhance endogenous glucocorticoid action on lung fluid balance and inflammation. DESIGN: Controlled laboratory study. SETTING: University research laboratory. SUBJECTS: Adult Sprague-Dawley rats (n = 66). INTERVENTIONS: Rats were intraperitoneally injected with carbenoxolone (2 x 10 mg.kg(-1).day(-1) for 3 days) and allowed free access to water and food. Rats were further challenged with endotoxin instillation (1 mg/kg). MEASUREMENTS AND MAIN RESULTS: We discovered that carbenoxolone significantly increased messenger RNA expression of all three epithelial sodium channel subunits in distal lung tissues (two-fold increase of alpha-subunit, four-fold increase of beta-subunit, and two-fold increase of gamma-subunit) as well as in trachea. Carbenoxolone increased the amiloride-sensitive alveolar fluid clearance significantly. When rats were further challenged by endotoxin instillation (1 mg/kg), pretreatment with carbenoxolone significantly inhibited endotoxin-induced increase in lung neutrophils as well as tumor necrosis factor-alpha and cytokine-induced neutrophil chemoattractant-1 concentrations in serum and bronchoalveolar lavage fluid. CONCLUSIONS: These beneficial effects of carbenoxolone on lung fluid balance and inflammation are very similar to those expected when glucocorticoids are introduced exogenously. We conclude that carbenoxolone increased the actions of endogenous bioactive glucocorticoids on lung cells by reducing local steroid breakdown. PMID- 15343022 TI - Adrenergic agents modify cerebral edema and microvessel ultrastructure in porcine sepsis. AB - OBJECTIVE: To investigate the effects of adrenergic agents on the cerebral response to sepsis. DESIGN: Prospective, randomized, controlled, experimental animal study. SETTING: Medical school research laboratories. SUBJECTS: Twenty eight middle white pigs (25-30 kg). INTERVENTIONS: Pigs were anesthetized, mechanically ventilated, and randomly assigned to one of the following groups: cecal peritonitis (n = 5), cecal peritonitis with dopexamine (n = 5), cecal peritonitis with dopexamine and the beta2-adrenergic receptor antagonist ICI 118,551 (n = 4), cecal peritonitis with methoxamine (n = 5), cecal peritonitis with dopexamine and methoxamine (n = 4), and sham-operated (n = 5). Sham-operated pigs were killed after laparotomy, and pigs with cecal peritonitis were killed 8 hrs after its induction. Samples of frontal cerebral cortex were taken immediately after death, processed for light and electron microscopy, and then subjected to morphometric analysis. MEASUREMENTS AND MAIN RESULTS: There was significantly more (p <.0005) cerebral perimicrovessel edema in pigs with cecal peritonitis (80.2 microm2 +/- 5.3 sem) than in sham-operated pigs (26.2 microm2 +/- 2.7 sem) and significantly less (p <.0005) perimicrovessel edema in dopexamine-treated pigs with cecal peritonitis (39.8 microm2 +/- 5.5 sem) than in pigs with cecal peritonitis alone (80.2 microm2 +/- 5.3 sem). There was no significant difference between the amount of perimicrovessel edema in pigs with cecal peritonitis treated with dopexamine plus ICI118,551 and pigs with cecal peritonitis alone. The mean cerebral microvessel endothelial cell cross-sectional area in methoxamine-treated pigs with cecal peritonitis (26.3 microm2 +/- 2.6 sem) was significantly greater than that in pigs with cecal peritonitis alone (16.3 microm2 +/- 2.1 sem, p =.008) or in sham-operated pigs (12.3 microm2 +/- 1.3 sem, p =.0005). CONCLUSIONS: Dopexamine protects against cerebral edema formation in sepsis by stimulation of beta2-adrenergic receptors, whereas the alpha1 adrenoceptor agonist methoxamine induces cerebral microvessel endothelial cell swelling. PMID- 15343023 TI - Translating family satisfaction data into quality improvement. AB - BACKGROUND: Improvement of clinical care requires measurement of key dimensions of health care quality and action based on these measurements. Families, data analysts, clinicians, and administrators all have important roles to play. OBJECTIVE: To outline an approach to the measurement and utilization of family satisfaction data so that these data can be translated into health care quality improvement initiatives. DESIGN: Using a synthesis of existing knowledge about translation of satisfaction data into improvement strategies, this approach starts with selecting and implementing a satisfaction survey that reflects the key processes, providers, and places for the delivery of critical care. The survey results can be expressed in a way that prioritizes the opportunities for improvement. A comparison of results across sites, or use of a performance importance grid, can assist in this prioritization process. High-priority items can then be addressed by the multidisciplinary intensive care unit team using a systematic, evidence-based approach to improvement that includes implementation strategies that have been proven to effectively change clinician behavior. PMID- 15343024 TI - Practice parameters for hemodynamic support of sepsis in adult patients: 2004 update. AB - OBJECTIVE: To provide the American College of Critical Care Medicine with updated guidelines for hemodynamic support of adult patients with sepsis. DATA SOURCE: Publications relevant to hemodynamic support of septic patients were obtained from the medical literature, supplemented by the expertise and experience of members of an international task force convened from the membership of the Society of Critical Care Medicine. STUDY SELECTION: Both human studies and relevant animal studies were considered. DATA SYNTHESIS: The experts articles reviewed the literature and classified the strength of evidence of human studies according to study design and scientific value. Recommendations were drafted and graded levels based on an evidence-based rating system described in the text. The recommendations were debated, and the task force chairman modified the document until <10% of the experts disagreed with the recommendations. CONCLUSIONS: An organized approach to the hemodynamic support of sepsis was formulated. The fundamental principle is that clinicians using hemodynamic therapies should define specific goals and end points, titrate therapies to those end points, and evaluate the results of their interventions on an ongoing basis by monitoring a combination of variables of global and regional perfusion. Using this approach, specific recommendations for fluid resuscitation, vasopressor therapy, and inotropic therapy of septic in adult patients were promulgated. PMID- 15343025 TI - What is taught, what is tested: findings and competency-based recommendations of the Undergraduate Medical Education Committee of the Society of Critical Care Medicine. AB - INTRODUCTION: Addressing an unexpected shortfall of intensivists requires early identification and training of appropriate personnel. The purpose of this study was to determine how U.S. medical students are currently educated and tested on acute care health principles. HYPOTHESIS/METHODS: A survey of critical care education with telephone follow-up was mailed to the deans of all 126 medical schools. Web site review of medical school curricula for critical care education was performed. Upon invited request, four members of the Undergraduate Medical Education Committee (UGMEC) reviewed 1,200 pool questions of step II of the U.S. Medical Licensing Examination (USMLE) given to graduating medical students for critical care content. Descriptive statistics are employed. RESULTS: Survey response rate was 49% and 88% by the second mailing with Web site review. Forty five percent of U.S. medical schools responding had formal undergraduate critical care didactic curricula averaging 12 +/- 3 hrs: 60% were elective, 60% taught in the 4th year. Eighty percent of clinical ICU rotations offered were elective. Sixty percent of schools taught 11 key critical care procedures in the 3rd or 4th year; 17% required them to graduate. Nineteen percent of Step II USMLE questions had critical care content; 58% dealt with pulmonary or cardiac disease. CONCLUSIONS: Graduating medical students are tested (and licensed accordingly) on critical care knowledge, despite an inconsistent exposure to the discipline in medical school. The UGMEC has drafted competency-based recommendations for acute health care delivery that encourage mandatory didactic and procedural critical care training. The UGMEC recommends that critical care rotations with didactic curricula be required for undergraduate education and that acute care procedural skills be an important component of these curricula. PMID- 15343026 TI - Biventricular assist device for scombroid poisoning with refractory myocardial dysfunction: a bridge to recovery. AB - We report the usefulness of biventricular mechanical circulatory support in a 36 yr-old woman with refractory myocardial dysfunction resulting from scombroid poisoning. DESIGN: Case report. SETTING: Medical and surgical university care units. PATIENT: A previously healthy 36-yr-old woman with severe myocardial dysfunction unresponsive to epinephrine (1.3 microg/kg/min) and dobutamine (18 microg/kg/min) after the ingestion of cooked fresh tuna. INTERVENTION: Implantation at day 3 of a biventricular assist device consisting of two paracorporeal pneumatic pumps set at 70 beats/min to reach an output of 5.6 L/min during 8 days. MAIN RESULTS: The biventricular mechanical circulatory assist device allowed weaning of the inotropic drugs, maintenance of end-organ function, and support of the patient until myocardial recovery. The patient was successfully explanted 11 days after ingestion. Cardiac function had totally recovered, but a stroke was noted. At 3-yrs follow-up, there was no cardiac or neurologic sequela. CONCLUSIONS: This report describes severe myocardial dysfunction secondary to scombroid poisoning and demonstrates the usefulness of a mechanical circulatory assist device as a bridge to recovery. PMID- 15343027 TI - Acute lung injury and mechanical ventilation: need for quality assurance. PMID- 15343028 TI - Volutrauma, barotrauma, and ventilator-induced lung injury: lessons learned from the animal research laboratory. PMID- 15343029 TI - Microcirculation in distress: a new resuscitation end point? PMID- 15343030 TI - Do cultural differences in communication and visiting result in decreased family desire to participate in decision making? PMID- 15343031 TI - Shared decision-making in the ICU: entering a new era. PMID- 15343032 TI - Improvement in coagulation markers with antithrombin--beneficial in severe sepsis? PMID- 15343033 TI - Risk stratification in the changing field of cardiac surgery. PMID- 15343034 TI - Alternative activated macrophage: a new key for systemic inflammatory response syndrome and sepsis treatment? PMID- 15343035 TI - Is low-dose vasopressin the new reno-protective agent? PMID- 15343036 TI - Hypothermia, sepsis, and the granulocytes: lessons to learn beyond the cytokines. PMID- 15343037 TI - Evidence-based approach to family care in the intensive care unit: why can't we just be decent? PMID- 15343038 TI - Further considerations regarding the effects of physical restraint in the intensive care unit. PMID- 15343040 TI - Adrenal insufficiency: the link between low apolipoprotein A-I levels and poor outcome in the critically ill? PMID- 15343042 TI - Endotracheal tube narrowing during mechanical ventilation. PMID- 15343043 TI - Obesity-related excess mortality rate in an adult intensive care unit: a risk adjusted matched cohort study. PMID- 15343044 TI - Hemodynamic monitoring in obese patients: a big issue. PMID- 15343045 TI - Standardized order forms and very high drug levels. PMID- 15343047 TI - Large right-to-left shunts are associated with both spinal and cerebral decompression illness. PMID- 15343049 TI - Are medical emergency teams picking up enough patients with increased respiratory rate? PMID- 15343051 TI - Airway management is a crucial area of critical care practice. PMID- 15343055 TI - Examining novel concepts of the pathophysiology of depression in the chronic psychosocial stress paradigm in tree shrews. AB - Despite decades of research on psychiatric disorders, the aetiology and precise biological mechanisms that underlie depressive diseases are still poorly understood. There is increasing evidence that psychiatric disorders not only have a neurochemical basis but are also associated with morphological alterations in central nervous neurons and/or glial cells. Antidepressants may act by restoring structure as well as function of neural networks, meaning that they may, as a fundamental principle, affect neural plasticity underlying normal brain functioning. To examine these novel concepts of the pathophysiology of depression and antidepressant medication we have carried out a series of experiments using the chronic psychosocial stress paradigm in male tree shrews, an animal model with a high validity for the pathophysiology of depressive disorders, in which the animals were treated with the tricyclic antidepressant compound clomipramine. We found that one month of stress reduced cell proliferation in the dentate gyrus, and decreased the total hippocampal volume. Gene transcription analysis revealed that, under these experimental conditions, expression of genes known to be involved in processes of cell differentiation is suppressed. These effects of social conflict on hippocampal cells, including gene transcription, and on the entire hippocampal volume could be counteracted by chronic treatment with the antidepressant clomipramine. Stress also induced a constant hyperactivity of the hypothalamic-pituitary-adrenal axis, and suppressed both motor and marking behaviour. These neuroendocrine and behavioural stress-induced changes were also re-normalized by clomipramine. PMID- 15343056 TI - Drug-induced neurobehavioral plasticity: the role of environmental context. AB - Repeated administrations of addictive drugs produce long-lasting changes in brain and behavior. However, drug-induced neurobehavioral plasticity is not a mere function of the neuropharmacological actions of drugs, but the result of complex drug-environment interactions. In the present review we summarize results obtained in a series of studies using an animal model of drug-environment interaction, showing that environmental context and past drug history interact to modulate the effects of amphetamine, cocaine and morphine on behavior, gene expression and structural plasticity. These findings may help shed some light on the conditions necessary for addictive drugs to enduringly alter brain and behavior. PMID- 15343057 TI - Windows of vulnerability to psychopathology and therapeutic strategy in the adolescent rodent model. AB - Adolescence comes in association with puberty, when maturation and rearrangement of major neurotransmitter pathways and functions are still taking place. The neurobiological processes occurring in the brain during this developmental period have been so far poorly investigated. Yet, it is during adolescence that some major neuropsychiatric disorders may become evident, including ADHD, schizophrenia, and drug abuse. Moreover, the age-related neurobehavioural plasticity renders adolescents particularly vulnerable to the consequences of psychoactive drug exposure. In this view, there is an increased likelihood that addiction will develop when psychoactive drug use starts early during adolescence. From all these observations adolescence emerges as a critical phase in development. In the present review, we focus on recent neurobiological characterization of adolescent rats and mice. As for vulnerability to addictive behaviour, nicotine exposure during adolescence dose-dependently down-regulated levels of AMPA GluR2/3 subunits in the striatum, suggesting a reduced neurobehavioural plasticity in adult subjects. Comparable exposure during adulthood had opposite effects. It was found consistently that exposure to nicotine during adolescence, but not similar exposure in the post-adolescent period, increased the expression of specific subunits of the acetylcholine receptor in adult rats, thus enhancing the reinforcing efficacy of nicotine in a self-administration paradigm. The present data identified a specific age-window, characterized by long-term effects on behavioural and neurochemical indexes, of vulnerability. With respect to potential therapeutic approaches in ADHD, we studied the adolescent spontaneously-hypertensive-rat (SHR) in an intolerance-to delay operant-behaviour paradigm. The model was further validated by the finding that impulsivity was reduced by chronic methylphenidate administration. Impulsive SHR animals were characterized by reduced cannabinoid CB1 receptor density in the prefrontal cortex. Interestingly, an acute cannabinoid agonist increased levels of self-control behaviour in these animals. The present data suggest that pharmacological modulation of the cannabinoid system might improve some behavioural anomalies seen in ADHD. In conclusion, modelling the adolescent phase in rats and mice appears to be useful for the investigation of determinants of vulnerability to addiction and to other early-onset neuropsychiatric disorders. PMID- 15343058 TI - Epigenetic control of neurobehavioural plasticity: the role of neurotrophins. AB - Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are two neurotrophins involved in the differentiation, growth and maintenance of selected peripheral and central populations of neuronal cells, during development and at adulthood. Furthermore, neuronal activity enhances expression and action of these neurotrophins, modifying synaptic transmission and connectivity. Neurotrophin production has been shown to be experience-dependent. In particular, during early developmental phases, experiences such as maternal deprivation or exposure to an enriched environment markedly affect NGF and BDNF levels. At adulthood, psychosocial stress has been shown to markedly alter NGF and BDNF levels, both in plasma and selected brain areas, including the hypothalamus and hippocampus. These results have been extended to humans, showing that NGF levels are enhanced by emotional stress induced by parachute jumping. Overall, these findings suggest a role of neurotrophins as factors mediating both short- and long-term effects of experience on brain structure and function. PMID- 15343059 TI - More questions than answers. Commentary on Fuchs et al., examining novel concepts of the pathophysiology of depression in the chronic psychosocial stress paradigm in tree shrews. PMID- 15343060 TI - Depressed neurogenesis and its role in the genesis of depression. Commentary on Fuchs et al., examining novel concepts of the pathophysiology of depression in the chronic psychosocial stress paradigm in tree shrews. PMID- 15343061 TI - Neural plasticity and the promise of novel pharmacotherapies for depression. Commentary on Fuchs et al., examining novel concepts of the pathophysiology of depression in the chronic psychosocial stress paradigm in tree shrews. PMID- 15343062 TI - Novel ideas about novelty. Commentary on Badiani and Robinson drug-induced neurobehavioral plasticity: the role of environmental context. PMID- 15343063 TI - Behavioural plasticity-induced changes in drug response. Commentary on Badiani and Robinson drug-induced neurobehavioral plasticity: the role of environmental context. PMID- 15343064 TI - Context gives meaning. Commentary on Badiani and Robinson drug-induced neurobehavioral plasticity: the role of environmental context. PMID- 15343065 TI - Blockade of D2 dopamine receptors in the VTA induces a long-lasting enhancement of the locomotor activating effects of amphetamine. AB - The present study examined the effects of pre-exposure to eticlopride, a D2 dopamine receptor antagonist, in the ventral tegmental area (VTA) on the subsequent locomotor activating effects of amphetamine (AMPH). Rats were pre exposed to one of three doses of eticlopride (0.75, 3.0 or 12.0 microg/0.5 microl per side) or saline (0.5 microl/side) in the VTA, once every third day, for a total of three infusions. Locomotor activity was recorded for 2 h following each pre-exposure injection. The low and intermediate doses of eticlopride produced no effects, while the high dose decreased locomotor activity compared to saline controls. 10-14 days following the last pre-exposure injection, all rats were challenged with AMPH (1.0 mg/kg, ip) and locomotor activity was recorded. Rats pre-exposed to the low dose of eticlopride exhibited enhanced locomotor activity whereas those pre-exposed to the intermediate or high doses did not differ from saline pre-exposed controls, suggesting that blockade of D2 dopamine receptors in the VTA can lead to sensitized locomotor responding to AMPH. To investigate the possible mechanism by which the low dose of eticlopride induced sensitization, extracellular levels of dopamine were measured as increasing concentrations of eticlopride (0.1, 1.0, 10.0 and 100.0 micromol/l) were perfused through a microdialysis probe implanted in the VTA. Only the lowest eticlopride concentration elevated extracellular dopamine levels. Therefore, as in the case of AMPH-induced sensitization, the induction by eticlopride of sensitization to AMPH may be initiated by the ability of eticlopride to increase extracellular levels of dopamine in the VTA. PMID- 15343066 TI - The GABAB agonist baclofen blocks the expression of sensitisation to the locomotor stimulant effect of amphetamine. AB - The purpose of the present study was to test the possible influence of baclofen, a GABAB agonist, on the long-term sensitisation to amphetamine in rats. As expected, chronic amphetamine treatment (1.5 mg/kg i.p. daily for 10 days) led to an increased locomotor response to amphetamine (0.75 mg/kg i.p.), when the animals were challenged 20 days after the end of repeated treatment. Baclofen (2 mg/kg i.p.), administered before the test session, did not significantly modify the spontaneous locomotor activity of rats, but decreased the normal and, to a greater extent, the sensitised locomotor response to amphetamine; thus baclofen prevented the expression of sensitisation to amphetamine. Moreover a previous chronic treatment with baclofen (2 mg/kg i.p. daily for 10 days) attenuated the amphetamine-induced locomotor activity in sensitised, but not in control animals. This effect was observed 20 days after the last baclofen administration. In conclusion, the present results demonstrate that GABAB receptors play an important role in the expression of the sensitised behavioural response to amphetamine and further support a potential role of GABAB agonists in the treatment of psychostimulant addiction. PMID- 15343067 TI - Influence of handling or aversive stimulation during rats' neonatal or adolescence periods on oral cocaine self-administration and cocaine withdrawal. AB - Adverse early life events may influence vulnerability for drug intake. The influence of handling or aversive stimulation during neonatal or adolescent periods on adult cocaine oral self-administration and withdrawal were investigated. Neonatal or adolescent rats were exposed to a modified unpredictable stress paradigm or handling for 10 days. When adults, oral cocaine was offered through the two-bottle choice paradigm for 30 days. Rats were submitted to the forced swimming test after cocaine withdrawal. Overall, there was a significant increase of cocaine choice throughout the days of cocaine consumption and an interaction between interventions and cocaine daily choice. Control rats started cocaine intake at a lower level and increased cocaine choice over time, while animals submitted to neonatal interventions started cocaine intake at higher levels of choice, with less increase in cocaine intake during the period of cocaine exposure. Rats receiving aversive stimulation during adolescence also started taking cocaine solution at higher levels. Significantly higher immobility duration and shorter latency to immobility during the forced swimming were detected in these same adolescents that received unpredictable stress, when compared to the control or handled rats, while there was no difference for rats stimulated neonatally. Therefore, early life events increase initial preference for cocaine and promote changes in its abuse pattern, according to the intensity of the event and the age of the individual at the time of the event. Moreover, adverse experiences during adolescence, but not in neonatal phases, increase the vulnerability to depressive-like behaviors during cocaine withdrawal of adult rats. PMID- 15343068 TI - The impaired coping induced by early deprivation is reversed by chronic fluoxetine treatment in adult fischer rats. AB - The symptoms of depression include feelings of reduced coping ability and increased helplessness. Early life adversity increases vulnerability to depression. In rats, the quantification of ability to cope with adverse challenge can be achieved using preexposure to an inescapable aversive stimulus and subsequent assessment of escape or avoidance deficits in the same environment. Here we investigated the predictive validity of a model in which, in the Fischer rat strain, postnatal isolation leads in adulthood to a state of increased sensitivity to develop an escape or avoidance deficit. On days 1-14 rat pups were isolated for 4 hours (early deprivation, ED) or for 15 minutes (early handling, EH), or were left completely undisturbed (non-handling, NH). In adulthood, subjects were placed in a shuttle box and half were exposed to brief, mild foot shocks (preexposure, PE) and the other half were non-preexposed (NPE). Half of the PE and NPE subjects were then treated for 21 days with fluoxetine and the other half with vehicle. In males, although there was no overall preexposure effect on avoidance behaviour, ED-PE and ED-NPE and EH-PE and EH-NPE demonstrated an avoidance deficit relative to NH. Fluoxetine attenuated this deficit and most notably in ED-PE. In females, vehicle ED-PE demonstrated an avoidance deficit relative to NH-PE; fluoxetine attenuated this ED effect. These findings provide supportive evidence for the predictive validity of this depression model. PMID- 15343069 TI - Nucleus accumbens dopamine receptors in the consolidation of spatial memory. AB - Nucleus accumbens dopamine is known to play an important role in motor activity and in behaviours governed by drugs and natural reinforcers, as well as in non associative forms of learning. At the same time, activation of D1 and D2 dopamine receptors has been suggested to promote intracellular events related to neural plasticity. Therefore, in this study we wished to investigate the role of the two classes of dopamine receptors within the nucleus accumbens on the consolidation of spatial information. On day 1, CD1 male mice were placed in an open field containing five different objects and, immediately after three sessions of habituation, the animals were focally injected within the nucleus accumbens with either the D1 antagonist SCH 23390 (12.5, 25 or 50 ng/side), or the D2 antagonist sulpiride (25, 50, 75 or 100 ng/side). Twenty-four hours later the ability of mice to discriminate an object displacement was assessed. Both the D1 and the D2 antagonists impaired the ability of mice to detect the spatial change. If the highest doses of the two antagonists were injected 2 h after the end of the last of the habituation sessions, no effect was observed in the reactivity to spatial change examined 24 h later. These data demonstrate that activation of both D1 and D2 receptors within the accumbens is necessary in the early stages of the consolidation of spatial information. The data are discussed in terms of involvement of nucleus accumbens dopamine in information processing in the absence of explicit reinforcers. PMID- 15343070 TI - Object recognition impairment in Fmr1 knockout mice is reversed by amphetamine: involvement of dopamine in the medial prefrontal cortex. AB - Fragile X syndrome is an X-linked form of mental retardation including, among others, symptoms such as stereotypic behaviour, hyperactivity, hyperarousal, and cognitive deficits. We hypothesized that hyperactivity and/or compromised attentional, cognitive functions may lead to impaired performance in cognitive tasks in Fmr1 knockout mice, the most widely used animal model of fragile X syndrome, and suggested that psychostimulant treatment may improve performance by acting on one or both components. Since hyperactivity and cognitive functions have been suggested to depend on striatal and prefrontal cortex dopaminergic dysfunction, we assessed whether amphetamine produced beneficial, positive effects by acting on dopaminergic corticostriatal systems. Our results show that Fmr1 knockout mice are not able to discriminate between a familiar object and a novel one in the object recognition test, thus showing a clear-cut cognitive impairment that, to date, has been difficult to demonstrate in other cognitive tasks. Amphetamine improved performance of Fmr1 knockout mice, leading to enhanced ability to discriminate novel versus familiar objects, without significantly affecting locomotor activity. In agreement with behavioural data, amphetamine produced a greater increase in dopamine release in the prefrontal cortex of Fmr1 knockout compared with the wild-type mice, while a weak striatal dopaminergic response was observed in Fmr1 knockout mice. Our data support the view that the psychostimulant ameliorates performance in Fmr1 knockout mice by improving merely cognitive functions through its action on prefrontal cortical dopamine, irrespective of its action on motor hyperactivity. These results indicate that prefrontal cortical dopamine plays a major role in cognitive impairments characterizing Fmr1 knockout mice, thus pointing to an important aetiological factor in the fragile X syndrome. PMID- 15343076 TI - Quick tips. Get the WOUND PICTURE. PMID- 15343077 TI - Skin IQ. Pouching a wound. PMID- 15343078 TI - ICD-9-CM coding changes impact wound care practices. PMID- 15343079 TI - The interdisciplinary CWS, 5 years later. PMID- 15343080 TI - What caused a "blood blister" to develop? PMID- 15343081 TI - Why won't some wounds heal? PMID- 15343082 TI - Assessing protective sensation with a monofilament. PMID- 15343084 TI - Hair-thread tourniquet syndrome in a cognitively impaired nursing home resident. PMID- 15343085 TI - Retrospective analysis of negative pressure wound therapy in open foot wounds with significant soft tissue defects. AB - OBJECTIVE: To compare treatment outcomes of open foot wounds with significant soft tissue defects using either negative pressure wound therapy (NPWT) or standard wet-to-moist saline-soaked gauze dressings. SUBJECTS: : Forty-seven subjects with open wounds with significant soft tissue defects. DESIGN: A retrospective analysis of sequentially selected records of patients treated with NPWT or standard wet-to-moist dressings. RESULTS: Risk of complications, subsequent foot surgeries, and hospital readmissions (secondary outcomes) were all reduced by 70% or more for the patients treated with NPWT. Length of stay during readmissions tended to be shorter; rates of wound cavity filling and wound healing (primary outcomes) tended to be greater with NPWT, although the differences were not statistically significant. CONCLUSION: Patients included in the study treated with NPWT had fewer foot-related complications, less additional foot surgery, and fewer readmissions for the study foot than did patients treated with standard therapy. Further research using randomized clinical trials is needed to confirm the findings of the current study's data. CLINICAL RELEVANCE: Wound cavity filling time and time to complete closure were similar in both study groups, despite the fact that patients receiving NPWT were older, had larger wounds, and had poorer nutrition. NPWT may be an effective modality for the healing of open wounds with significant defects and for reducing subsequent care. PMID- 15343087 TI - Wound assessment: tips and techniques. PMID- 15343088 TI - Incorporating laboratory values in chronic wound management. PMID- 15343090 TI - [After the "era of antibiotics", the "era of resistance"]. PMID- 15343091 TI - [Toxoplasmosis seroconversion in pregnant women. The differing attitudes in France]. AB - INTRODUCTION: Since 1992 France has been running a toxoplasmosis prevention program based on the serological detection of infections during pregnancy. In the absence of a consensus, the seroconversions discovered are managed in different ways, varying from one centre to another. OBJECTIVE: To describe the habits of the specialised centres in France and propose means to reduce the heterogeneity. METHODS: A survey using a questionnaire mailed to the centres of parasitology in France specialised in the management of toxoplasmosis seroconversion during pregnancy. RESULTS: All the 30 centres surveyed replied. Five of them do not provide recommendations for treatment and were excluded from analysis. The attitudes of the 25 other centres varied greatly with regard to the indications for therapeutic abortion and amniocentesis, treatment protocols with pyrimethamine and sulfamides, as well as in the frequency of sonographical monitoring. CONCLUSION: In the absence of National guidelines, the management of seroconversions discovered during the prenatal prevention of congenital toxoplasmosis program is left to the discretion of the specialised centre. This results in variations from one town to the next. This heterogeneity underlines the lack of knowledge on the most efficient and acceptable means of preventing and treating congenital toxoplasmosis. In order to improve this situation, the French parasitologists have launched a program of consensual meetings aimed at harmonising the treatment protocols and identifying the points that require further studies. PMID- 15343092 TI - [Non-justified visits to emergency units. Proposal of differentiated care]. AB - OBJECTIVE: The number of visits to emergency units in public hospital settings in France increases every year. The adequation between admission to an SAU--Service d'accueil d'urgence (emergency unit) and the clinical status of the patient must be checked to improve handling upstream of the SAU. METHOD: A prospective study was conducted in the SAU of the University Hospital in Nantes to assess the proportion of patients who would benefit from direct hospitalisation, scheduled in a department of specialised or polyvalent medicine. RESULTS: This proportion was of 10%. Seventy-three percent of the patients were aged over 60. They were referred in 77% of cases by their treating physician and in 10.4% of cases by the physician on duty. Thirty-three percent of cases were non-specified organ diseases, 20% were dermatological affections, 12% broncho-pulmonary infections and in the same proportion rheumatological pathologies; other affections were rare. COMMENTS: The results of this study must be confirmed in a pilot study in which the general practitioner would refer any patient, that he would have sent to an SAU, directly to a medical department without passing through the SAU. To do so, using a cell phone, the practitioner would contact the hospital physician who would find a hospital bed. The impact of this new modality of hospitalisation on the SAU could be assessed in terms of the number of admissions avoided to the SAU. PMID- 15343093 TI - [Typology of the French medical learned societies. Survey of 129 organizations]. AB - INTRODUCTION: A study on the relationship between the ANDEM--Agence Nationale pour le Developpement de l'Evaluation Medicale (French agency for the development of health technology assessment) and the learned societies showed that a definition of the role of these organisations and criteria to define learned societies were lacking. We conducted a survey among the learned societies in the field of medicine so as to elaborate a definition. METHODS: We used the files of the learned societies of the Anaes--Agence Nationale d'Accreditation et d'Evaluation en Sante (French Agency for accreditation and evaluation in health) in 1998, which included 225 organisations. We sent a letter together with a single-page questionnaire and copies of the 2 publications on the relationship between the ANDEM and the learned societies. To analyse the suggestions for a definition, having read the replies, a segmentation of the population, the means and modalities of action and the aims and fields of action were used. A proposal for a definition was discussed during 2002/2003 with the board of the Federation des Specialites Medicales (FSM) (Federation of medical specialties) regrouping 33 medical and 12 surgical societies. Three meetings between a representative of the Anaes and the board of the FSM resulted in the proposition of so-called "validation" criteria for learned societies. A search on the French Internet listing such societies was made using a research motor (March 20, 2004). RESULTS: Out of the 225 organisations contacted, 129 (57%) replied. Among the latter, 115 considered themselves as a learned society and 14 did not. Among the 115 organisations defining themselves as a learned society, 97 proposed a definition, 16 made use of the definitions proposed in the questionnaire and 81 proposed their own definition. The analysis identified 6 important elements (reporting, knowledge, education, research, diffusion, promotion). The data analysed permitted the elaboration of a definition based on the proposals of the learned societies: "An organised group, within the framework of a given discipline, the members' aim of which is to report on their work, to improve knowledge on their discipline, to ensure education and research, to diffuse information on their work and research and to support and promote their discipline". Thirty-two of the 45 societies of the FSM accepted the 11 validation criteria proposed, some of which were commented on. Only one society abstained and 12 did not reply. No French Internet web site listing the learned societies specified a definition of criteria for inclusion in the lists. DISCUSSION: A definition of the learned societies and validation criteria, obtained through the participation of the principle actors, could serve as a basis for the recognition and identification of such societies. PMID- 15343094 TI - [The risk of rabies in France and the illegal importation of animals from rabid endemic countries]. AB - INTRODUCTION: Since October 2001, France has the status of a country free from rabies regarding warm-blooded animals. Nevertheless, the risk of rabies is ever present and related to the illegal importation of animals from countries in which rabies is considered to be endemic. OBSERVATION: In 2001, a puppy imported illegally from Morocco died from rabies in Gironde, requiring the prophylactic treatment of 21 persons exposed and the euthanasia of a contact animal, as well as the application of legal measures against the couple at the origin of the importation. CONCLUSION: Illegal importations of animal from areas of rabid enzootics are frequent. It is important to inform all the tourists who visit these countries on the sanitary risks involved, in terms of human and animal health, in bringing such animals back to their own country. PMID- 15343095 TI - [Extended medullary necrosis revealing a homozygote sickle-cell anaemia]. PMID- 15343096 TI - [Pulmonary sequestration]. PMID- 15343097 TI - [Taking into account their pain]. PMID- 15343098 TI - [The elaboration of analgesic protocols for the elderly. The experience of a multidisciplinary group]. AB - TAKING INTO CONSIDERATION THE PAIN: This has become a right for the patient, an ethical and deontological necessity for the physician, and a near-judicial obligation for care centres. The current regulations authorise nurses to apply analgesic treatments, within the framework of written protocols, signed and dated by the physician. A NEW METHOD: In order to determine the main acute pains of the elderly in institutions and to develop analgesic protocols permitting rapid and efficient relief, with priority given to non-medical treatments, a multidisciplinary group consisting of nurses and non-nursing care workers has been made up. Training tools have been created and distributed. CONCRETE RESULTS: After 14 months' work, the initial objective has been reached. Five analgesic protocols taking into account acute pain in the elderly have been drawn-up. All the care workers have been trained by their nursing colleagues. Following validation, the protocols have been applied for the past year in the department and are now being circulated to other care structures. PMID- 15343099 TI - [Morning stiffness]. AB - STIFFNESS AND UNSTRETCHING: Morning stiffness in the elderly is a painful entity often ignored, but frequent in institutions. Unstretching in the morning is part of the criteria in the diagnosis of inflammatory pathologies, but morning stiffness in old patients appears different from that described in rheumatological diseases and arthrosis. FROM AN AETIOLOGICAL POINT OF VIEW: Various affections can be accompanied by morning stiffness in an elderly patient: arthrosis, but also various consequential affections, static disorders, deformity. We felt it was important to define the precise criteria that individualise this pathology so as to improve its management. MANAGEMENT: Non medicinal measures often reduce the intensity and duration of this symptom and implicate all those attending to the patient. Medicinal treatment has a more limited place in the fragile elderly patient, but its role remains to be assessed. PMID- 15343100 TI - [Headaches in the elderly]. AB - ACCORDING TO AGE: It is generally thought that the prevalence of headaches decreases with ageing. However recent studies, with stricter epidemiology and methodology, clearly indicate that this decreases is less obvious than that perceived. PRIMARY AND SECONDARY HEADACHES: In elderly patients, primary headaches and notably migraine (often with altered presentation) are less frequent, even though new authentic cases may appear. However the other types of headache are clearly present. In particular, the incidence and prevalence of secondary headaches slightly increases and they represent up to 30% of all the headaches observed, compared with less than 10% in young or adult patients. The causes of such symptomatic headaches are multiple and it is important to be able to identify them since an aetiological treatment is often possible. REGARDING TREATMENT: The symptomatic treatment of headaches in the elderly follows the same principles as that of younger patients. However, the side effects and drug interactions related to pharmacological treatments can be disastrous in fragilised patients. It is therefore important to emphasize the interest of physical methods of analgesia, particularly adapted to the elderly. Some of these methods have demonstrated their efficacy and are recommended by international consensuses. PMID- 15343101 TI - [Physical analgesic therapy in the elderly. Its interest in the management of headaches]. AB - A FREQUENT AND DISABLING AFFECTION: Headaches in elderly patients are far more frequent that what is generally presumed; their prevalence is estimated of between 25 and 54% after the age of 70. They must be treated because of the potential impact on the patients' mental, functional and social status. NON MEDICINAL ANALGESIC TREATMENTS: Various treatments have demonstrated their efficacy on different types of headache and are recommended in the international consensuses. This is notably the case with electrotherapy, cryotherapy, massage and biofeedback the correct use by physiotherapists of which leads to a decrease in the pain, duration and frequency of the attacks, and improves the quality of life of these patients. OTHER PHYSICAL TREATMENTS: Techniques such as acupuncture, relaxation, manual therapy or postural training are less documented but can be proposed. Most of them, alone or in supplement to medicinal treatment, are particularly adapted to elderly patients and limit the need for drugs, the side effects and interactions of which may have severe consequences in fragile patients and those suffering from multiple diseases. PMID- 15343102 TI - [The Brugada syndrome. To be evoked in case of malaise in a young adult]. AB - A DIAGNOSIS TO BE EVOKED: The Brugada syndrome is a rare but serious inherited disease that causes sudden cardiac death by ventricular tachycardia and fibrillation, especially in younger men. Diagnostic problems are related to the possible absence of symptoms although the electrocardiogram (ECG) reveals the characteristics of a Brugada syndrome and to the variations in the ECG in the same patient over time. THREE ELECTROCARDIOLOGICAL ASPECTS: Type 1 corresponds to the historical description with ST segment elevation at point J of at least 3 mm from its summit and upward convex ST segment followed by a negative T wave. In Type 2, the extent of point I is of 2 mm, the ST segment has a saddle form and remains at least 1 mm above the isoelectric line, the T wave is positive or biphasic. In Type 3, the terminal section of the ST segment never exceeds 1 mm above the isoelectric line. In the case of a Type 1 ECG, a pharmacodynamic test is of no interest. REGARDING TREATMENT: The only treatment to have demonstrated its efficacy is the implantable automatic defibrillator, indicated in symptomatic patients. PMID- 15343104 TI - [The management of a genital ulcer 2/2]. PMID- 15343105 TI - [Profession: vascular doctor. Inaugural address of the President]. PMID- 15343107 TI - [Vascular resistances in lower limb arterial disease used to evaluate the quality of peripheral arteries. Preliminary study]. AB - Peripheral resistance of lower limb arteries is not a common clinical measurement despite easy-to-apply non-invasive techniques. The aim of our trial has been to test this new parameter as a marker of arterial lesions. MATERIAL AND METHODS: In this prospective study, hemodynamic data acquired non-invasively [Ankle brachial index (ABI), peak systolic velocity (PSV), end-diastole velocity (VTD) and peripheral artery quality] in patients with lower limb arterial disease were compared with the resistance index measured in the common femoral artery (Arbeille index: IR=VTD/PSV). RESULTS: Between February 1, 2003 and April 30, 2003, 150 measurements of resistance index were made in 99 individuals with ischemia following a strain (69% men, 31% women, average age 64.8+/-15.2 years). Forty-one of the resistance measurements (27%) were also made in patients with arterial disease factors (diabetics, renal replacement therapy). The resistance index was closely associated with end-diastole velocity. (r=-0.76; DDL=148; p<0.01) as well as peripheral artery quality (r=0.81; DDL=148; p<0.01); on the contrary the resistance index was not affected by the arterial disease factors (0.3+/-0.1 vs 0.33+/-0.12; p=NS). CONCLUSION: These findings suggest that the hemodynamic impact might be evaluated more precisely diabetics and renal replacement therapy patients whose hemodynamic parameters are difficult to assess due to their arterial disease factors. Prospective studies are needed to determine the role of the resistance index in the follow up of these patients, and its contribution in comparison with the measurement of end-diastole velocity. PMID- 15343108 TI - [Management of venous thromboembolic disease in ambulatory and hospital settings: an assessment of general practices in France]. AB - OBJECTIVE: In spite of increasing use of prophylactic treatments, venous thromboembolic disease (VTD) remains an important public health problem because of its high morbidity and mortality, in particular after major orthopedic surgery. The aim of this study is to provide an assessment of current medical management of VTD and post-thrombotic disease (PTD) in France. METHOD: A random sample of 210 investigators including hospital practitioners (orthopedic surgeons and anesthesiologists), general practitioners (GP), and angiologists working in the ambulatory setting completed a standardized questionnaire about their current medical practice. RESULTS: VTD led to an additional 8.5 days of hospital stay needed for the management of deep vein thrombosis (DVT) and 15.8 days in the event of pulmonary embolism (PE). Doppler diagnosis of DVT was obtained in 97% to 100% of patients. When PE was suspected, a lung scan was performed in 62% of patients managed by hospital practitioners and in 43% of patients treated by GPs. Whereas the rate of hospitalization for distal DVT diagnosed in outpatients did not exceed 14% of patients, the rate of hospitalization for proximal DVT reached 40%-48%. Eighty-eight percent of the patients with PE were managed in a hospital setting. Two-thirds of the patients with PE were given low-molecular-weight heparin for an average duration of 9.3 days and one-third of patients were treated with non-fractioned heparin for an average duration of a 11 days. Doppler diagnosis of PTD was established in 98% of patients and 98% of the patients with PTD were treated with elastic contention stockings. CONCLUSION: Based on physicians' report of their current medical practice, this study provides useful data on diagnostic and therapeutic practices in the management of VTD and PTD. PMID- 15343109 TI - [Therapeutic education of patients receiving anticoagulants for thromboembolic venous disease: description of the Educ'AVK program]. AB - Educ'AVK is an education program designed for patients starting an oral anticoagulant treatment following a thromboembolic event (deep venous thrombosis or pulmonary embolism). Patients enrolled are judged capable of self-management of their drug treatment. The intervention, adapted to a short-term treatment, consists in an 3-step one-on-one teaching session conducted by a trained pharmacist. 1) Identification of the patient's needs--or educational diagnosis. 2) Definition of educational objectives adapted, first, to the patient's cognition: to be able to describe indications for therapy and potential risks factors for anticoagulant bleeding, to interpret INR lab tests and to solve a problematic treatment situation related second, to the patient's behaviour: to be able to anticipate the risk (how to enhance compliance, how to deal with a missed dose.), to take relevant decisions when dealing a therapeutic incident (bleeding, INR outside of the targeted range). 3) Choice of appropriate teaching contents and methods to reach the objectives. We used the "individual guidance" technique associated with original tools: a picture book to describe the pathology and the treatment; a booklet synthesizing all the information given by the educator during the consultation. This booklet presents 3 key-points: the information is specifically targeted to thromboembolic venous disease; the information integrates the risk-level of the patient (3 different booklets according to the patient's level of risk, of bleeding, of thromboemboly, no specific risk); there is a section where the patient writes down his/her INR results in a table specifically adapted to his/her risk level--bleeding, thromboemboly, no specific risk--and giving recommendations for the management of INR out of the targeted range. PMID- 15343110 TI - [Impact of an education program on patient behaviour favoring prevention of drug related adverse events: a pilot study in patients receiving oral anticoagulants for thromboembolic venous disease]. AB - OBJECTIVE: Adverse events related to oral anticoagulants represent a major public health problem. Including patient education as part of the prevention strategy could contribute to improved effectiveness and safer use of drugs. The present study aimed at evaluating the outcomes of a patient education program inspired by recommendations from French Health Authorities (AFSSAPS) and based on an "individual guidance" approach. METHOD: The study was conducted in two groups of hospitalized patients treated with oral anticoagulants for thromboembolic disease. Each patient in the first (intervention) group attended an individual teaching session conducted at discharge by a trained pharmacist. Patients in the second (control) group were given usual care. These two groups were compared at inclusion (before intervention) and three months later. The outcomes considered were the acquisition of: 1) knowledge, 2) risk anticipation and compliance behaviours characterized by the stability of INR and the incidence of hemorrhagic episodes during the period of observation. RESULTS: Fifty-nine patients (average age 65 years) were included (29 in the intervention group and 30 in the control group). Three months after the intervention, the intervention group exhibited 1) better knowledge (higher rate of restitution of treatment-related information- name of the drug, administration plan, targeted range for INR (...), interpretation of INR results (p<0.05), management of a specific scenario where INR declines concomitant to elevation of anticoagulant dose (p<0.05)); 2) higher rates of relevant behaviours (p<0.05)--in the event of a missed dose, anticipating an event with a high risk of bleeding, dealing with signs of overdose--and higher compliance profile (ns) (stability of INR, and number of hemorragic episodes). A multivariate model integrating the potential explanatory variables for frequency of hemorrhagic episodes at 3 months (demographic data, history of thrombotic disease, INR stability, reference group (intervention/control)), showed that the only variable significantly associated with frequency of bleeding events was the reference group of the patient (p=0.05; odds-ratio=4.5, interval of confidence: [1-21]). CONCLUSION: Multivariate analysis demonstrated that the probability of developing a hemorrhagic event when taking an oral anticoagulant is on average 4-fold greater in patients given usual care than in patients given individual guidance a pharmacist. A larger randomized trial is currently under way in the Rhone-Alpes region, France, to validate these exploratory results. PMID- 15343111 TI - [Intracardiac thrombus in Behcet's disease. A report of three cases]. AB - Intracardiac thrombus formation is exceptional in Behcet's disease (less than 50 cases had been reported). Among 180 patients with Behcet's disease (according to the criteria of the international study group for Behcet's disease), three were diagnosed as having intracardiac thrombus. All were male, the mean age at the time of the diagnosis of the intracardiac thrombus was 27 years and the mean disease duration was 4.8 years. The presenting symptom was hemoptysis in two patients and heart failure in the third. Surgery was the first attempted treatment in two patients but thrombus recurred and resolved after medical treatment. The third patient was given corticosteroids and anticoagulants as the first line treatment. The thrombus did not resolve and the patient declined surgery. Intracardiac thrombus formation should be considered when a young Behcet's disease patient presents with an intracardiac mass. Medical treatment including corticosteroids, immunosuppressive drugs and anticoagulants should be considered as the first line treatment and surgery should be considered when there is no resolution of the thrombus or when it becomes massive and extensive. In certain cases, thrombolytic treatment becomes an interesting alternative to surgery. PMID- 15343112 TI - [Arteriovenous fistula of the renal vessels due to septic embolism in a patient with bacterial endocarditis]. AB - A 27 year-old woman developed acute pain of the right flank during the course of an infectious endocarditis. A septic arteriovenous fistula of the renal vessels of a solitary functioning kidney was demonstrated. The cardiac valvular lesions required a prosthetic aortic and mitral replacement valves. An attempt to occlude the fistula by embolization with several coils was unsuccessful and led to surgery: extracorporeal repair enabled complete closure of the fistula. During the long-term follow-up, the aortic prosthetic valve had to be changed. Renal function was satisfactory and remained stable over time. Renal arteriovenous fistula is an exceptional complication of bacterial endocarditis despite the frequency of septic emboli during the course of the disease. PMID- 15343113 TI - [Gadolinium-medium filter-protected percutaneous stended carotid angioplasty]. AB - We report on a patient who underwent a percutaneous carotid angioplasty with stenting. The procedure was protected by a filter and used gadolinium instead of iodine. No neurological or renal complication occurred. This observation represents one of the first publications describing the use of this contrast medium in a complex neuroradiological intervention. PMID- 15343114 TI - [Reversed carotid filter for the prevention of distal emboli during an iliac recanalization]. AB - We describe the use of a carotid filter in preventing distal emboli during the recanalization of an occluded left iliac stent two months earlier. The filter was reversed and introduced through an ipsilateral 6 French sheath along its 0.014" guidewire and placed against the occluded stent. The stent is recanalized using a hydrophilic guidewire controlaterally and a new stent was inserted into the old one. The filter was retracted into the introducer by pulling cut the special 0.014 guidewire, with a 0.018" diameter tip, and the whole system was removed via the left common femoral artery. Many emboli were found in the removed filter. We noted that the recanalization of an occluded stent causes more distal embolization than the native iliac artery. This technique allows placing the filter between the embolic lesion and the distal arterial territory, without passing through the lesion, especially since fibrinolysis may not be efficient in 2-months-old occlusions. PMID- 15343115 TI - [Cyclosporine 2% eye drops preparation]. AB - PURPOSE: Cyclosporine administration is very effective in the case of immunological diseases of the cornea, conjunctive or uvea. Moreover, it is widely used in the case of high-risk rejection corneal transplantation. We present a preparation of cyclosporine 2% eye drops. METHODS: Cyclosporine 2% eye drops are prepared following a particular formulation including one part commercially available cyclosporine oral solution (Sandimmun) diluted in four parts of sterile castor oil. Manufacturing procedures maintain the sterile state of the preparation with a laminar airflow hood placed in a particulate controlled room, with pharmacists, technicians and clerical personnel wearing sterile clothes. Physical and chemical monitoring during and after manufacture for each batch guarantees the process and minimizes the risk of batch rejection. Chemical analysis of cyclosporine is conducted using a validated stability-indicating high performance liquid chromatographic assay (reverse-phase). Blood dosages taken after the first administration at the 24th hour (after administration of the 6th drop) check for systemic integration. RESULTS: Cyclosporine 2% eye drops are fairly stable: 12 months after manufacturing, concentrations result in levels not statistically different from concentrations measured the day of preparation. After a daily regimen of six drops in the eye, cyclosporine 2% eye drops have a very low systemic bioavailability, because the blood concentrations only reach the detection limit of the fluorescence polarization immunoassay used for cyclosporine drug monitoring. This explains the absence of systemic toxicity. CONCLUSION: Cyclosporine 2% eye drops can be available in the hospital pharmacy. The eye drops are stable at room temperature and can be delivered up to 12 months after manufacture. No local adverse effects have been noted, probably in relation with the very low concentration of ethanol in the ocular preparation. PMID- 15343116 TI - [Abnormal eyelid positions in Brown syndrome]. AB - PURPOSE: To describe the possible abnormal upper eyelid positions in congenital Brown syndrome (BS) and discuss physiopathology. CONTEXT: A typical symptom of BS, the incapacity to raise the eye in adduction, is usually accompanied by a retraction of the upper eyelid. However, as the abnormal position of the eye often refers to a patient looking straight ahead into the distance, abnormal eyelid positions are possible but are on the whole unknown. PATIENTS AND METHODS: The study investigated 82 patients with unilateral congenital BS, who consulted between 1989 and 2001. The primary position of the upper eyelid concerned was carefully measured, each side separately. The patients were then classified into three groups according to the type of difficulty encountered when raising the upper eyelid: group 1--those with a raising deficiency in adduction (42 cases), group 2--those with a raising deficiency in adduction and in primary position (26 cases), group 3--those with a global raising deficiency with predominance in adduction (14 cases). Fifty patients were operated on for strabismus, after which the position of the upper eyelid was remeasured. RESULTS: An abnormal position of the upper eyelid was noted in 23 patients (28%) and the distribution between the different groups was as follows: group 1, 14%; group 2, 30%; group 3, 63%. The following malpositions were found: pseudoptosis with hypotropia, eight cases; ptosis, six cases; pseudoretraction with vertical oculomotor disorder, five cases; real upper eyelid retraction, four cases. Ptosis and pseudoptosis were observed more often when BS involved the nondominant eye; retraction or pseudoretraction was observed more often when BS involved the dominant eye. After surgery for vertical strabismus, the abnormal position of the upper eyelid had remarkably decreased and only three cases required specific surgery for ptosis. CONCLUSION: The abnormal position of the upper eyelid in primary position was found in nearly one-third (28%) of the patients with BS, therefore a quite frequent occurrence. It sometimes involved associated congenital ptosis. However, the most frequent symptom was a deficiency of the eyelid that was caused either by the patient being incapable of preventing the eyelid from falling (ptosis and pseudoptosis) or the patient making a compensatory effort to raise the eyelid (retraction and pseudoretraction). PMID- 15343117 TI - [Prismatic treatment in low-vision rehabilitation of patients with age-related macular degeneration]. AB - PURPOSE: To evaluate visual improvement in an age-related macular degeneration (ARMD) population with bilateral central scotomas, after a 3- to 6-month prismatic treatment. PATIENTS AND METHODS: Prisms prescribed for both eyes were oriented according to eccentric fixation. Criteria were visual acuity (VA), reading ability, contrast sensitivity subjective visual comfort, prism wearing frequency and autonomy level. RESULTS: Thirty-one patients (23 women, 8 men) 65 95 years old (mean, 78 years) were selected. Mean long distance VA remained stable (0.249 at inclusion and 0.261 after prismatic treatment); long distance VA was unchanged for 51.6% of patients, improved by at least one line for 35.5% and decreased for 12.9%. Reading ability was slightly improved (average of 1 line), as was short distance VA (P14 at inclusion, P12 after prismatic treatment); reading ability was unchanged for 45.2% of patients, improved by at least one line for 38.7%, and decreased for 16.1%. Contrast sensitivity was improved for all levels: to 98% of contrasts (mean at inclusion 0.248 and 0.280 after prismatic treatment), to 50% of contrasts (0.214 at inclusion and 0.238 after prismatic treatment) and to 25% of contrasts (0.157 at inclusion and 0.185 after prismatic treatment). Visual comfort improved immediately after prismatic treatment for 90.3% of patients; 80.6% of patients wore their prismatic correction regularly. DISCUSSION AND CONCLUSION: Prismatic treatment in this population mainly improves contrast sensitivity and visual comfort. It provides patients with better initiation and stabilization of their eccentric viewing. Most of the time, prismatic treatment must be associated with low-vision rehabilitation. PMID- 15343118 TI - [Failure of photocoagulation therapy for diabetic macular edema]. AB - PURPOSE: To analyse clinical and tomographic features of diabetic macular edemas (DME) unresponsive to laser photocoagulation. METHODS: Retrospective study of 43 consecutive eyes presenting with severe DME defined by a macular thickening 450 microm or more, involving the fovea, and associated with either cystoid spaces and/or hard exudates threatening the center of the macula. All 43 eyes underwent subthreshold grid laser therapy (a mean of 2.3 sessions). Mean follow-up was 21 months. RESULTS: Photocoagulation was beneficial in 39% of the eyes (n=17) and failed in 60% (n=26). Comparing these two groups, we found that some tomographic and clinical signs were strikingly more frequent in the unresponsive eyes than in the responsive ones. OCT findings reported foveal thickening greater than 700 microm (38%), subclinical serous foveal detachment (65%), subclinical vitreomacular interface abnormalities (19%) and as clinical signs, 44% of the unresponsive eyes exhibited rapid progression of retinal ischemia preceding ME constitution. CONCLUSION: Long-standing diffuse DME carries an unfavorable prognosis. In cases of unresponsive DME exhibiting these tomographic and clinical signs, alternative therapies such as intravitreal injections of steroids and/or vitrectomy with posterior hyaloid removal should be considered, instead of iatrogenic and useless iterative grid laser treatments. PMID- 15343119 TI - [Sleep-related breathing disorders and normal or high-tension glaucoma: 35 patients with polysomnographic records]. AB - PURPOSE: To look for early stage glaucoma symptoms in patients referred for sleep related breathing disorders or sleep apnea syndrome. PATIENTS AND METHOD: Thirty five patients between October 2001 and April 2002 consecutively admitted for polysomnographic evaluation were investigated. Each patient had complete ophthalmologic examination before the nocturnal record. A blue-on-yellow Humphrey computerized perimetry test was given, and a standard perimetry with a color vision test if abnormalities were found. The criteria studied were intraocular pressure, optic nerve cupping and aspect, visual field indices and visual field aspect, which were compared to the respiratory disturbance index. Twenty-seven subjects were included in this analysis, nine women and 18 men. RESULTS: In eight patients, the respiratory disturbance index was higher than 10. All intraocular pressures were normal. Visual field defects were found in two patients with both standard and blue-on-yellow perimetry, and in 12 patients with only blue-on yellow perimetry. Mean deviation and visual field aspect correlated well with the respiratory disturbance index (p=0.026, p=0.033). Other visual field indices were not correlated with the respiratory disturbance index. CONCLUSION: In this study, we found visual field alterations with blue-on-yellow computerized perimetry that did not exist with standard perimetry. The visual field defects were more frequent when the respiratory disturbance index was poor. PMID- 15343120 TI - [Orbital meningoencephalocele: two case studies]. AB - Primitive orbital meningocele is a rare congenital malformation. It is defined as a herniation of meninges into the orbit through a congenital defect in the orbital bones. In meningoencephalocele, there is atrophic brain tissue in the herniated meningeal sac. We report the cases of two babies who presented with a medial orbital tumoral syndrome. Computed tomography showed a cystic tumor close to a bony defect in the anterior frontoethmoidal junction. A transcranial approach confirmed the diagnosis of meningoencephalocele; it was resected and the bony and dural defects were closed. The outcome was uneventful. Orbital meningoencephalocele is probably linked to an abnormal closure of the rostral neuropore. It generally presents in the first months of life with very suggestive radiological and clinical features. Surgical treatment using a transcranial approach is indicated in all cases and provides definitive recovery. PMID- 15343121 TI - [Poor visual outcome in acute multifocal placoid pigment posterior epitheliopathy. A clinical case]. AB - We report a case of a fifty-year-old women suffering from acute multifocal posterior placoid pigment epitheliopathy (AMPPPE), very atypical in terms of clinical appearance and course. From this observation, we provide a progress report on the usual course of AMPPPE as well as atypical features of this affection through reports in the literature; finally we detail disorders ressembling both AMPPPE and serpiginous choroidopathy, the main differential diagnosis. PMID- 15343122 TI - [Hereditary retinal pathologies and AMD: new therapeutic perspectives]. AB - This article summarizes our current knowledge on two types of retinal diseases: hereditary retinal degeneration and wet aged-related macular degeneration. Our understanding of retinal physiopathology in hereditary retinal degeneration and the successful experimental therapeutic results on animal models call for a new approach to these patients to prepare future clinical trials. Longitudinal follow up of the functional alteration rhythm based on international standards and by expert centers is an essential prerequisite to including these patients in future clinical trials. Creating international databases that include data on follow-up using electrophysiological, psychophysical and morphological analyses would require standards defining how each of these procedures should be carried out. Furthermore, the relevance and value of the various examinations would then be evaluated in a longitudinal manner. The repeated use of these procedures in the various centers would bring to light any limitations these techniques may have for use in prospective studies. Continuous re-evaluation of these investigative techniques will therefore be necessary, a crucial factor in the preparation of multicenter clinical studies. The inclusion of patients phenotyped at different centers would require that certification procedures be set up for these centers. A great leap forward, clinical trials on new antiangiogenic approaches for the treatment of neovascular AMD are currently underway. The reasons for the switch between the neovascular and atrophic forms of AMD are as yet unknown, but these new approaches are based on the events that occur sequentially during the angiogenic response. PMID- 15343124 TI - [Examination and surgical indications for blepharoplasty]. AB - Cosmetic blepharoplasty has become very popular these last years and can easily be performed with local anesthesia. Its benefits are numerous and side effects minimal. Nevertheless, the different types of eyelid impairment are numerous and the patient is often not aware of them. This paper discusses the importance of the clinical examination targeting proper diagnosis of the type of eyelid impairment so as to recommend the most adapted surgical technique. The different types of cosmetic eyelid impairment are presented. PMID- 15343125 TI - [Upper cosmetic blepharoplasty and the eye brow]. AB - Upper cosmetic blepharoplasty allows to treat cutaneo-orbicularis excess of the upper eyelid, which is often associated with fat prolapse. Using the same surgical way, it is also possible to treat external heaviness of the brow, mild brow ptosis and glabellar wrinkles. We will describe the most important stages of these different surgical techniques. PMID- 15343126 TI - [Lower lid blepharoplasty]. AB - Baggy lower eyelids result from herniated orbital fat, excess skin, hypertrophic or lax orbicularis sometimes associated with lower lid laxity resulting from degeneration of the lateral canthal tendon with age. Two main approaches are used: skin-muscle flap or the transconjunctival approach in younger patients who have herniated orbital fat with minimal or not excessive lower eyelid skin. Preoperative evaluation of the patient is extremely important in lower lid blepharoplasty before any surgical procedure: excessive skin, herniated orbital fat, retraction and laxity must be evaluated. In our practice, a lateral canthoplasty is often performed in case of horizontal lower laxity in order to avoid round eye, scleral show or lower lid ectropion. In some patients, subperiosteal elevation of malar soft tissue is associated with lower blepharoplasty via subciliary incision. PMID- 15343127 TI - [Complications of blepharoplasty]. AB - Blepharoplasty complications are infrequent, most often minor and transitory, rarely major and permanent with functional or esthetic consequences. Treatment is above all preventive: screening at risk patients with a history of ophthalmic problems, but also general illnesses that would contraindicate blepharoplasty. Patients must be informed of possible risks through informative booklets with the most important points underlined. Complications can be purely ophthalmological, the more serious sequelae being partial or complete visual loss due to ischemic optical neuropathy, with very poor prognosis, or more rarely compression of the ocular globe by intraorbital hemorrhage, which has a better prognosis provided the origins are quickly recognized and treated immediately. Other visual complications include oculomotor problems, keratoconjunctivitis sicca, epiphora, and chemosis of lymphatic origin. Eyelid complications are more frequent: ptosis on the upper eyelid or lagophthalmia caused by incorrect resection of the skin, scarring and eyelid fold anomalies. The most serious esthetic complication is the malposition of the lower eyelid, which can manifest as retraction, lagophthalmia, ectropion, deformation of the external canthus, or lower eyelid tissue relaxation. These malpositions are quite often minor, sometimes reversible, but at times major, with psychological, esthetic and functional consequences that are difficult for the patient. Other local complications also arise: enophthalmia with a sunken lid, as well as under- and overcorrection. General complications can include scarring related to pigmentation problems and residual hematomas, and exceptionally infections going as far as the orbital fat tissue. Finally, other complications are related to new laser surgical techniques that are responsible for ectropion of the lower eyelid and even burns or residual redness, or complications related to periocular injections of filling material. A comprehensive review of prevention, diagnosis, and management of complications after blepharoplasty is presented. PMID- 15343128 TI - [Palpebral pseudoedema: rare etiologies]. PMID- 15343129 TI - Magnetic resonance imaging (MRI) and arthro-MRI in the evaluation of the chondral pathology of the knee joint. PMID- 15343130 TI - Evaluation of patellar chondromalacia with MR: comparison between T2-weighted FSE SPIR and GE MTC. AB - PURPOSE: To compare two different MR sequences to tissue signal suppression in the study of patellar cartilage abnormalities. MATERIALS AND METHODS: We examined 26 patients with magnetic resonance (MR) imaging: sequences included spectral presaturation with inversion recovery (SPIR), with fat suppression and T2 weighted images, magnetization transfer contrast (MTC) sequences, T1-weighted and T2-weighted spin-echo sequences. All patients underwent conventional knee arthroscopy and in all patients a hyaline cartilage lesion was assessed in three articular zones: the patellar medial facet, the lateral facet and the patellar crista. Was assessed 78 articular facets. The lesions were classified using a standard arthroscopic grading system adapted to MR imaging: normal cartilage that corresponds to the grade 0 according to the Noyes grading system, low grade lesions that correspond to the grade I e IIa and high grade lesions that correspond to grades IIb and III. The arthroscopic results were compared with MR images. We assessed the MR diagnostic accuracy, sensitivity, specificity and MR positive predictive value and negative predictive value of the two sequences taking into consideration total lesions, and high-grade and low grade lesions separately. RESULTS: Twenty-four low grade lesions (16 grade I e 8 grade IIa) and 18 high grade lesions (10 grade IIb e 8 grade III) were diagnosed by arthroscopy. Regarding low grade and high-grade lesions together, the accuracy was 77% for MTC sequences and 90% for SPIR sequences. In identifying low-grade lesions, the sensitivity was 88% for SPIR sequence and 42% for MTC sequences. Specificity for the detection of all lesions was 89% for the SPIR sequences and 94% for the MTC sequences. The SPIR sequence visualised water content abnormalities in degenerating cartilage, which are representative of low-grade lesions. The sensitivity of the sequence enabled us to obtain improved contrast for detecting cartilage surface irregularities. The MTC sequences allowed us to grade high grade lesions susceptible to surgery and small cartilage defects in the presence of joint fluid. The MTC sequences were insufficient in the diagnosis of early stages of chondromalacia because the suppression of the signal of bonded water reduced the contrast among areas of articular cartilage with different water content. For this reason cartilage oedema and early superficial fibrillation were not identified. CONCLUSIONS: In our experience the SPIR sequence proved superior to the MTC sequence in the identification of low grade lesions of the patellar cartilage. The overall value of such sequences in the study of articular pathology also needs to be assessed in the others sites where the articular cartilage is thinner and surfaces more curvilinear. PMID- 15343131 TI - Association between monoclonal gammopathy of undetermined significance (MGUS) and diffuse idiopathic skeletal hyperostosis (DISH). AB - PURPOSE: To determine whether an association exists between monoclonal gammopathy of undetermined significance (MGUS) and diffuse idiopathic skeletal hyperostosis (DISH). MATERIALS AND METHODS: Radiological studies of skeleton were performed in a series of 76 patients (51 males and 25 females) affected by MGUS, observed in a consecutive period of 36 months. They revealed hyperostotic lesions similar to those described in DISH. RESULTS: Patients affected by MGUS complicated by DISH were 35 (24 males and 11 females). The prevalence of skeletal hyperostosis (46%) appears higher than that reported by the literature in the general population (mean 15 %). Typically, the vertebral column is the predominant site of abnormalities in MGUS as well as in DISH: involvement of dorsal spine was observed in 16 males and 10 females; cervical spine in 12 males and 4 females, and lumbar in 9 males and 2 females. Peripheral ossifying enthesopathy ("whiskering" in the pelvis) was found in 7 cases, 5 males and 2 females. CONCLUSIONS: DISH is probably an ossifying diathesis of unknown aetiology; its occurrence in multiple myeloma and especially in MGUS is much higher because it is a fortuitous association. Many causes (accidental, dysmetabolic or degenerative) of DISH-like syndrome complicating dysgammaglobulinemias have been supposed, but at the present time this association remains a mere guess. A pathogenetic "event" responsible for hyperostosis may be supposed: in fact, in MGUS bone stimuli induced by osteoclast activating factor (OAF) and plasma cell proliferation -- even if lower than those observed in multiple myeloma -- are much more prolonged in time (see the slow progression of this condition, before it becomes symptomatic), so that the rate of calcium mobilized from skeleton and localized into soft tissues (i.e., tendons and ligaments) is higher. In this manner, the incidence of DISH-like ossification is higher than that observed in multiple myeloma and in the general population. PMID- 15343132 TI - Hip election radiographic plane (HERP) in the preoperative planning of prostheses with femoral neck preservation. AB - In the preoperative radiographic planning of total hip prosthesis implantation, the hip election radiographic plane (HERP) is an essential, "dedicated" tool, i.e. it is indispensable for correct implant planning, especially as regards the femoral component and specifically some types of devices which, like the Biodynamic or CFP prostheses, allow preservation of the femoral neck. PMID- 15343133 TI - High-frequency ultrasound in the study of male breast palpable masses. AB - PURPOSE: To evaluate the role of high frequency colour-Doppler ultrasound in the study of diffuse or local breast nodules in males, and compare these results with those of mammography. MATERIALS AND METHODS: We studied 105 men aged between 12 and 82 years (mean age 42.3 years) with a palpable breast mass with or without pain. All patients underwent clinical and ultrasound examination. Seventy-eight also underwent mammography, whereas 27 did not because of young age (under 25 years) (n=10), no clinical or sonographic suspicion of a malignant mass (n=16), and ulcerated neoplastic lesion (n=1). The final diagnosis derived from surgery in six patients and from three-year follow-up for 99 patients. RESULTS: Eighty nine patients had gynecomastia (85%), nine had adipomastia (8%), one had fibrolipoma (1%), five had carcinoma (5%) (invasive ductal carcinoma in four and bifocal ductal carcinoma and lobular carcinoma in one patient) and one had liposarcoma (1%). The clinical examination detected a bilateral (n=66) or unilateral (n=39) breast mass. Ultrasonography provided the correct diagnosis of diffuse nodular pathology in all cases and orientated diagnosis towards malignancy (6 cases) or benignity (1 case) of the lesions. No additional information was obtained from mammography, as compared to high-frequency ultrasonography. The integration of colour-Doppler in the examination was of little use as it demonstrated extensive vascularisation of most (5 cases) of the malignant lesions. CONCLUSIONS: Our results indicate that ultrasound allows the detection and characterization of palpable breast masses as well as correct local staging of neoplasms by identifying the degree of infiltration of the surrounding tissues. PMID- 15343134 TI - Magnetic resonance with manganese-DPDP (mangafodipir) of focal solid pancreatic lesions. AB - PURPOSE: To assess the accuracy of magnetic resonance (MR) with Mangafodipir (Mn DPDP) in the identification of focal solid pancreatic lesions. The possibility of lesion characterisation based on quantification of Mn uptake was also investigated. MATERIALS AND METHODS: Thirty-four patients (11 females, 23 males, aged 21-75, mean age 57) selected at sonography (US) and spiral CT (SCT) for clinically suspected focal solid pancreatic lesion, were studied between June 2000 and July 2001. Patients eligible for surgery underwent MR imaging (1.5 T) before and after infusion of 5 micromol/kg of Mn-DPDP (0.5 ml/kg Teslascan, Nycomed Amersham Health, Oslo, Norway). The baseline examination included FSE T2 weighted sequences and fat-saturated and non fat-saturated breath-hold gradient echo T1-weighted sequences (FMP SPGR). After Teslascan infusion, fat-saturated and non fat-saturated FMP SPGR sequences were repeated. Thirty of the 34 patients had a definitive diagnosis provided by reference standards such as post-operative histology (22 cases), cytology (FNAB) and/or a follow-up period of at least 6 months (8 patients). As regards lesion characterisation, the signal-to-noise ratio (S/N ratio) and contrast-to-noise ratio (C/N ratio) of lesions compared to the pancreatic parenchyma were calculated using ROIs before and after contrast infusion. RESULTS: The definitive diagnosis was pancreatic malignancy in 18 patients, focal pancreatitis in 5 and neuroendocrine tumours in 3. Four patients with suspected lesions at US and/or SCT were free of focal pancreatic disease. Mn DPDP MR identified 17/18 malignancies, 2/3 endocrine neoplasms 5/5 focal pancreatitis; the 4 patients with no pancreatic lesions were correctly identified. The Mn-DPDP MR accuracy in detecting focal pancreatic solid lesions was 93%. MR missed 1 small adenocarcinoma (the only pT1 in our group) and 1 insulinoma (with Mn uptake similar to the surrounding parenchyma). Due to the small number of inflammatory lesions included in the study, no significant differences were found in signal intensity and Mn-DPDP uptake between focal pancreatitis and neoplasms. CONCLUSIONS: MR with Mn-DPDP is very accurate in the identification of focal solid pancreatic lesions. Mangafodi-pir is also very useful for excluding the presence of pancreatic lesions suspected at US or CT. The characterisation of lesions, in particular of inflammatory versus neoplastic lesions, remains problematic and requires further investigation. PMID- 15343135 TI - Comparison between the site of multislice CT signs of gastrointestinal perforation and the site of perforation detected at surgery in forty perforated patients. AB - PURPOSE: To compare the site of multislice spiral computed tomography (MSCT) signs of gastrointestinal perforation and the site of perforation at surgery in forty perforated patients. MATERIALS AND METHODS: Between January 1 and July 31, 2003, a total of 40 patients (23 men and 17 women) underwent surgery for gastrointestinal perforation. In all cases, plain radiography of the abdomen was integrated by MSCT with the following parameters: 0.5 seconds gantry rotation time, 2.5-5.0 mm slice thickness, 3.75 reconstruction interval, 120 kV, 250-300 mAs, pitch 1.5, after intravenous administration of 140 ml of contrast agent at 3 ml/s with an automatic injector and a delay time of 70 seconds from the injection of the contrast agent. The MSCT findings were: free air and free fluid observed in supramesocolic compartments and/or in inframesocolic compartments and bowel wall discontinuity. The sites of the MSCT findings were compared with the site of perforation observed at surgery. RESULTS: Free air was detected in 60%, free intraperitoneal fluid in 92.5%, and a combination of both findings in 57.5% of the 40 cases examined. There were no cases of bowel wall discontinuity. In nine patients with gastroduodenal perforation, free air and free fluid were detected in combination and free air was localised in supramesocolic compartments in all cases; in two patients with jejunal perforation, free intraperitoneal fluid was observed both in supramesocolic and inframesocolic compartments; in six patients with acute perforated appendicitis, free air was never detected, while free fluid was observed in all cases in inframesocolic compartments; in six patients with isolated sigmoid perforation free air was observed in four cases in supramesocolic compartments while free fluid was seen in both supramesocolic and inframesocolic compartments. CONCLUSIONS: MSTC is the most reliable diagnostic method with which to assess gastrointestinal perforation as it allows detection of even small amounts of free air in the abdomen, which are a sign of perforation. In our study, the comparison of the sites of MSCT signs of perforation with those observed at surgery showed that in gastroduodenal perforations free air and free fluid are present in combination and free air is localised in supramesocolic compartments; in acute perforated appendicitis free air is absent, while free fluid is present in inframesocolic compartments; in isolated sigmoid perforations free air, if present, is localised in supramesocolic compartments, while free fluid is seen in both compartments. PMID- 15343136 TI - Abdominal ultrasonography and chest radiography are of limited value in the emergency room diagnostic work-up of severe trauma patients with hypotension on the scene of accident. AB - PURPOSE: To evaluate the reliability of chest radiograph (CR) and abdominal ultrasonography (US) performed in the Emergency Room (ER) in identifying life threatening thoracic or abdominal lesions in a group of severely injured patients, who developed arterial hypotension immediately after a trauma. MATERIALS AND METHODS: The results of all abdominal US and CX performed in the ER in severe blunt trauma patients, with on-the-scene systolic blood pressure 90 mmHg, from November 2000 to November 2002, were analysed. When these initial investigations failed to identify a possible cause of hypotension, a computed tomography (CT) of the chest and abdomen was obtained. RESULTS: Overall, 54 patients were studied; twenty-two patients (40.8%) were hypotensive both on accident scene and on the arrival at the ER and 32 (59.2%) were hypotensive on accident scene, but not in the ER. Forty-five patients had an US in the ER, in 3 patients it was inconclusive, whereas 2 other patients died before the confirmatory investigations could be performed. Among the remaining 40 patients, 11 had a hemoperitoneum (HP) hat was diagnosed by US in 7 cases and missed in 4. Twenty-nine patients had no HP and their US was negative in 24 cases and positive in 5; the US had a sensibility of 63.6% and a specificity of 82.8% The CR was obtained in the ER in 39 patients and it was able to identify 6 pneumothoraces (PNX) out of 20 and 2 hemothoraces (HT) out of 17; the sensitivity for PNX and HT was 30.0% and 11.8%, respectively. One patient with an aortic dissection had a normal CX. CONCLUSIONS: Both CX and US are not reliable to identify possible PNX, HT and HP in hypotensive trauma patients and can delay the treatment of life threatening conditions. In these patients, a CT of the torso is warranted. PMID- 15343137 TI - Virtual endoscopy in odontogenic sinus disease. Study technique and main pathological findings. AB - The use of CT scans in dental pathology is an established technique. The potential applications of Dentascan are further enhanced by the use of virtual navigation software, resulting in endoscopy-like imaging of the maxillary sinus, thus optimising both the diagnostic and therapeutic approach to sinus pathology of dental origin. The aim of this paper is to illustrate the technical methodological aspects of maxillary sinus virtual endoscopy with Dentascan software and to document the most important and frequent diseases. PMID- 15343138 TI - Magnetic resonance imaging in simple snoring and obstructive sleep apnea-hypopnea syndrome. AB - PURPOSE: Obstructive Sleep Apnea-Hypopnea Syndrome (OSAHS) is a condition characterised by periodic cessation of breathing during sleep, associated with Upper Air-Digestive Ways (UADW) morphologic abnormalities that can be detected, in awake patients, by using various imaging techniques. The purpose of this study is to determine the usefulness of MR imaging and new original morphometrical measurements that we are proposing in patients with Sleep Obstructive Breathing Disordered (SOBD). MATERIALS AND METHODS: We studied 70 patients (52 with OSAHS and 18 snoring without OSAHS) using 1.5T and 0.5T MR imagers with neck and head coils and T1-DP-T2-weighted SE sequences. During the procedure, the patients were awake and with tidal breathing. We also evaluated sagittal pharyngeal diameters at different levels; the length and maximum width of soft palate; the distance between the hyoid bone and the C2C3-Me line (ideally joining the geometrical centre of the C2-C3 intervertebral space to the lower point of mandibular symphysis) measured on the perpendicular; the angle resulting from the longitudinal axis of the cervical spine and the epiglottis axis (alpha); the slope angle of the tongue -- resulting from the longitudinal axis of the cervical spine and the longitudinal axis of the tongue (beta). We used sagittal, coronal and axial sections of the head and neck. RESULTS: In OSAHS patients, pharynx calibre medium sizes were reduced compared with simple snoring patients. Only in OSAHS patients (not in simple snorers without OSAHS) we observed: 16 patients with narrowing sites = or <3 mm. On axial images we observed three different narrowing patterns: rounded, with greater anterior-posterior axis; with greater axis in lateral direction. In OSAHS patients we also observed, on average, increase of the distance between the hyoid bone (Hmr point) and the line C2C3-Me; increase in the angle resulting between cervical rachis and epiglottis (alpha); reduction of sloping angle of the tongue (beta). CONCLUSIONS: MR imaging, together with the morphometrical measurements we are proposing, is useful to evaluate UADW in SOBD. In particular, we noted that increase of the distance between the hyoid bone (Hmr point) and the line C2C3-Me (due to lowering of the hyoid bone), increase in the angle resulting between the cervical rachis and the epiglottis (alpha) and the reduction of the sloping angle of the tongue (beta), are highly specific and sensitive indexes in OSAHS. There are different levels and findings of narrowing in OSAHS and their identification is very important for a surgical approach: the uvulo-palato-pharyngoplasty (UPPP) has a higher success rate in patients with obstruction at retro-palatopharynx site, but it is associated with no (or poor) results in hypopharyngeal obstruction. PMID- 15343139 TI - Percutaneous treatment of carotid stenosis with protection system: preliminary experience. AB - PURPOSE: To assess the feasibility, safety, and efficacy of carotid artery stenting with filter device. MATERIALS AND METHODS: Between May 2001 and July 2002, a total of 96 consecutive patients (100 lesions) who presented with symptomatic >70% diameter stenosis, asymptomatic >90% diameter stenosis or post endarterectomy as well as post-radiotherapy hemodynamically significant stenosis underwent carotid artery stenting with filter protection at our institute. The mean age of the patients was 72 (range 51-91). There were 22 female (23%), and 74 male (77%). Of the 96 patients, 62 patients (65%) did not filled the NASCET surgical criteria and 24 (26%) were classified as ASA 4. All the patients underwent pretreatment with antiplatelet agents. All procedures were performed with a standardized monitoring system by a multidisciplinary team. Filtration system for cerebral protection was consistently used. Outcome measures were procedural atheroembolic events, including all-stroke and death rates up to 30 days. RESULTS: A protection system type "TRAP" (Microvena, Italy) or a protection system type "EPIFILTER" (Boston Scientific, USA) were employed in 28% and 67% of cases, respectively. In the remaining 5% it was not possible to cross the stenosis with the filter. The stent placement was achieved in 95 of the procedures (95%). In 86% of cases a primary stenting technique was performed, with balloon pre-dilation of the lesion and subsequent stent advancement through the stenosis accounting for 9% of cases. At the time of protection system removal, 21 filter (21%) showed presence of macroscopic particles. We recorded 8 (8%) homolateral neurological events: two major strokes, one minor stroke and five TIAs (all the patients with TIA fully recovered within 2 hours). General complications (5%) included one lesion of the median nerve, two groin haematomas (one with disability for the patient, and one requiring surgery), one myocardial infarct and one lower limb ischaemia requiring surgical intervention. All major complications (4%) (two major strokes, one myocardial infarct and one lesion of the median nerve) occurred within the first 31 cases, the 69 following procedures not showing any neurological or non-neurological major events. At 30 days all patient were still alive. CONCLUSIONS: Filter protection during carotid artery stenting seems technically feasible as well as reasonably safe and effective. The EPIFILTER system turned out to be of easier employment due to its "monorail" system and because it does not usually need a catheter of dedicated recovery. Further studies in larger groups of patients are required to best characterize criteria guiding the choice for the right protection system. PMID- 15343140 TI - Subintimal angioplasty of infrapopliteal artery occlusions in the treatment of critical limb ischaemia. Short-term results. AB - AIM: To evaluate the feasibility and efficacy of subintimal infrapopliteal angioplasty (SIA) as a method for recanalization of occluded tibial arteries in the treatment of critical limb ischaemia (CLI). MATERIALS AND METHODS: Between January 2002 and September 2003, 20 patients with CLI were submitted to SIA; of these, 16 had diabetes mellitus. All patients had foot ulceration or gangrene and ten had rest pain. All patients were treated with SIA of one or more vessels of the popliteal district. Overall, thirty-four arteries of the infrapopliteal district underwent revascularization; in 9 cases, SIA of superficial femoral artery occlusions was associated. Technical success was evaluated on angiography at the end of the procedure: revascularization of at least one of the 3 leg vessels with re-establishment of arterial flow to the foot was regarded as a technical success. Pain relief (when pain was present) and healing of foot ulceration, without above-the-ankle major amputation (limb salvage), were defined as clinically successful. During the follow-up (mean: 9 months; range: 6-21 months) all patients were checked 6 months after the procedure by clinical examination and colour-Doppler ultrasound. RESULTS: The technical success rate of SIA in the revascularization of the infrapopliteal vessels was 85%. In the 17 technically successful cases, pain had entirely resolved in 9/10 cases and trophic lesions of the foot healed in 14/17 cases. In this group, 9 patients underwent minor amputation; 2 underwent major above-the-ankle amputation; one underwent to surgery 20 days after the SIA and required a femoro-tibial by-pass. In the 3 cases of technical failure (15%), revascularization of the entire occluded tract could not be achieved. Of these, one patient subsequently underwent major amputation. Nine months after SIA, the cumulative limb salvage rate was 85% (17/20 clinically successful cases) and the survival rate was 90%. Colour-Doppler US at 6 months showed 70% primary patency. No major complication occurred during the procedure. Five minor complications in four patients were managed endovascularly or healed spontaneously. CONCLUSIONS: SIA is a feasible and effective technique for foot revascularization in patients with CLI. Long occlusions or diffusely calcified arteries are suitable indications. Technical failure does not preclude conventional surgery. In patients treated with SIA, the risk of major amputation is low and mortality rate is nil. Minor complications can be managed using endovascular techniques. PMID- 15343141 TI - CT guided percutaneous fine needle biopsy of small lung lesions in outpatients. Safety and efficacy of the procedure compared to inpatients. AB - PURPOSE: To compare the safety and efficacy of CT-guided fine needle biopsy (FNAB) of small (<15 mm) lung lesions in inpatients and outpatients. MATERIALS AND METHODS: 108 consecutive inpatients (69 M, 39 F, mean age 56) and 121 consecutive outpatients (90 M, 31 F, mean age 50) who underwent CT-guided FNAB of small lung lesions were included. Lesion size, depth, number of needle passes, presence of emphysema were recorded. 22 G Chiba needles and the roll-over technique were used for all patients; if no significant pneumothorax was detected after FNAB, outpatients were allowed to go home and instructed to return in case of complications. The incidence of pneumothorax and other complications, sensitivity, specificity, diagnostic accuracy were calculated. RESULTS: 12 inpatients and 33 outpatients were lost to follow-up. No statistical differences were observed in lesion size, depth, needle passes, presence of emphysema between the groups. We had 15 pneumothoraces in inpatients, 4 requiring a chest tube, 12 in outpatients, 2 requiring a tube. Diagnostic accuracy was 92.7% in inpatients and 90.9% in outpatients. There were 7 false negatives in inpatients and 8 in outpatients, with negative predictive value of 79% and 78%, respectively. There were no false positives. All differences are nonsignificant. CONCLUSIONS: CT guided FNAB of small lung lesions is an equally safe and effective procedure in inpatients and outpatients; outpatient performance of FNAB can decrease costs. PMID- 15343142 TI - Evaluation of the dose to pediatric patients undergoing micturating cystourethrography examination and optimization of the examination. AB - PURPOSE: The aim of our work was to estimate the dose to paediatric patients undergoing micturating cystourethrography (MCU), and to optimize those examination procedures that were found to be particularly significant from the point of view of the radiation dose. At the same time it was also decided to evaluate the absorbed dose to the parents, who frequently assist the child during MCU. MATERIALS AND METHODS: The study was carried out on 220 children undergoing micturatig cystourethrography, by measuring in vivo the dose equivalent entering and exiting from the patient with thermoluminescent dosimetry and the KAP (Kerma Area Product). From the latter, the imparted energy and the average absorbed dose to the patients were subsequently calculated. RESULTS: The average absorbed dose was 0.69+/-0.54 mGy, with a variation interval of 0.126-3.110 mGy. A remarkable dispersion of the absorbed dose values was observed, also for subjects of similar size. This indicates that radiation protection of paediatric patients still allows for wide margins of optimization. In fact it was possible to verify that the dose depends not only on the size of the child, but also on his/her degree of cooperation, and on the technical ability of the operator. The latter aspect was estimated by comparing the doses delivered by two different physicians; differences by a factor of 5 were observed. The application of additional filtration of 2 mm of Al (total 5 mm Al) concurred to a dose reduction without significant variations in image quality. The effective dose received by the parents present during the examination was 4+/-7 microSv. CONCLUSIONS: A specific feature of paediatric MCU is not only the amount of radiation dose, often not negligible, but also the remarkable variation of the dose values, even within the same age group. The specialist has an important role in the justification of the examination: the adoption of a specific protocol has allowed reduction of the number of children undergoing MCU. Optimization of the examination requires quality control on the equipment, as well as the adoption of good radiographic techniques and the provision of suitable environments for children. The present study has helped to focus the attention of practitioners on radiation protection and, consequently, to reduce the dose delivered to the children. The effective dose received by the parents showed that their presence can be justified provided that they are adequately protected. PMID- 15343143 TI - Surveillance for fatal and nonfatal injuries--United States, 2001. AB - PROBLEM/CONDITION: Each year in the United States, an estimated one in six residents requires medical treatment for an injury, and an estimated one in 10 residents visits a hospital emergency department (ED) for treatment of a nonfatal injury. This report summarizes national data on fatal and nonfatal injuries in the United States for 2001, by age; sex; mechanism, intent, and type of injury; and other selected characteristics. REPORTING PERIOD COVERED: January-December 2001. DESCRIPTION OF SYSTEM: DESCRIPTION OF THE SYSTEM: Fatal injury data are derived from CDC's National Vital Statistics System (NVSS) and include information obtained from official death certificates throughout the United States. Nonfatal injury data, other than gunshot injuries, are from the National Electronic Injury Surveillance System All Injury Program (NEISS-AIP), a national stratified probability sample of 66 U.S. hospital EDs. Nonfatal firearm and BB/pellet gunshot injury data are from CDC's Firearm Injury Surveillance Study, being conducted by using the National Electronic Injury Surveillance System (NEISS), a national stratified probability sample of 100 U.S. hospital EDs. RESULTS: In 2001, approximately 157,078 persons in the United States (age adjusted injury death rate: 54.9/100,000 population; 95% confidence interval [CI] = 54.6-55.2/100,000) died from an injury, and an estimated 29,721,821 persons with nonfatal injuries (age-adjusted nonfatal injury rate: 10404.3/100,000; 95% CI = 10074.9-10733.7/ 100,000) were treated in U.S. hospital EDs. The overall injury-related case-fatality rate (CFR) was 0.53%, but CFRs varied substantially by age (rates for older persons were higher than rates for younger persons); sex (rates were higher for males than females); intent (rates were higher for self harm-related than for assault and unintentional injuries); and mechanism (rates were highest for drowning, suffocation/inhalation, and firearm-related injury). Overall, fatal and nonfatal injury rates were higher for males than females and disproportionately affected younger and older persons. For fatal injuries, 101,537 (64.6%) were unintentional, and 51,326 (32.7%) were violence-related, including homicides, legal intervention, and suicide. For nonfatal injuries, 27,551,362 (92.7%) were unintentional, and 2,155,912 (7.3%) were violence related, including assaults, legal intervention, and self-harm. Overall, the leading cause of fatal injury was unintentional motor-vehicle-occupant injuries. The leading cause of nonfatal injury was unintentional falls; however, leading causes vary substantially by sex and age. For nonfatal injuries, the majority of injured persons were treated in hospital EDs for lacerations (25.8%), strains/sprains (20.2%), and contusions/abrasions (18.3%); the majority of injuries were to the head/neck region (29.5%) and the extremities (47.9%). Overall, 5.5% of those treated for nonfatal injuries in hospital EDs were hospitalized or transferred to another facility for specialized care. INTERPRETATION: This report provides the first summary report of fatal and nonfatal injuries that combines death data from NVSS and nonfatal injury data from NEISS-AIP. These data indicate that mortality and morbidity associated with injuries affect all segments of the population, although the leading external causes of injuries vary substantially by age and sex of injured persons. Injury prevention efforts should include consideration of the substantial differences in fatal and nonfatal injury rates, CFRs, and the leading causes of unintentional and violence-related injuries, in regard to the sex and age of injured persons. PMID- 15343144 TI - Cholera epidemic associated with raw vegetables--Lusaka, Zambia, 2003-2004. AB - Zambia experienced widespread cholera epidemics in 1991 (13,154 cases), 1992 (11,659), and 1999 (11,327). In response to the large outbreak in 1999, the Zambian Ministry of Health (ZMOH) urged use of in-home chlorination with the locally produced solution, Clorin, and the practice increased substantially Clorin had been introduced in Zambia in 1998 as part of the Safe Water System (SWS), a point-of-use water disinfection and safe-water storage strategy launched by the Society for Family Health, in partnership with ZMOH, the U.S. Agency for International Development, and CDC. Although no outbreaks were reported during 2000-2002, cholera remained endemic. Epidemic cholera returned to Zambia in November 2003, when cases of toxigenic Vibrio cholerae O1, serotype Ogawa, biotype El Tor were confirmed in the capital city, Lusaka. During November 28, 2003-January 4, 2004, an estimated 2,529 cholera cases and 128 cholera deaths (case-fatality rate [CFR] = 5.1%) occurred in Lusaka. In February 2004, the Lusaka District Health Management Team (LDHMT) invited CDC to assist in an investigation of the epidemic. This report summarizes the results of that investigation, which implicated foodborne transmission via raw vegetables and demonstrated a protective role for hand washing with soap. The results underscore the importance of hygiene, clean water, and sanitary food handling for cholera prevention. PMID- 15343145 TI - Suspension of rotavirus vaccine after reports of intussusception--United States, 1999. AB - On July 16, 1999, CDC recommended that health-care providers suspend use of the licensed rhesus-human rotavirus reassortant-tetravalent vaccine (RRV-TV) (RotaShield, Wyeth Laboratories, Inc., Marietta, Pennsylvania) in response to 15 cases of intussusception (i.e., a bowel obstruction in which one segment of bowel becomes enfolded within another segment) among infants who received RRV-TV. The Vaccine Adverse Event Reporting System (VAERS) monitored for adverse events following licensure of RRV-TV on August 31, 1999. After the recommendation to suspend use of the vaccine, no additional cases were reported. This report describes the surveillance activities used to identify this vaccine adverse event, the emergency response, and follow-up investigations. Suspension of RRV-TV after the initial cases of intussusception parallels the removal of the Broad Street pump handle in response to John Snow's epidemiologic studies; both were decisive, life-saving public health actions. PMID- 15343146 TI - Outbreak of aflatoxin poisoning--eastern and central provinces, Kenya, January July 2004. AB - In May 2004, CDC Kenya, trainees of the CDC-supported Field Epidemiology and Laboratory Training Program (FELTP) in Kenya, the World Health Organization, and CDC were invited by the Kenya Ministry of Health (KMOH) to participate in the investigation of an outbreak of jaundice with a high case-fatality rate (CFR) in the districts of Makueni and Kitui, Eastern Province. Preliminary laboratory testing of food collected from the affected area revealed high levels of aflatoxin, suggesting that the outbreak was caused by aflatoxin poisoning, as was a previous outbreak in the same area in 1981. In the United States, aflatoxin concentrations are limited to 20 parts per billion (ppb), a level also adopted by Kenyan authorities. The 2004 outbreak resulted from widespread aflatoxin contamination of locally grown maize, which occurred during storage of the maize under damp conditions. Urgent replacement of the aflatoxin-contaminated maize with noncontaminated maize proved to be a critical intervention; however, as of July 21, a limited number of new cases continued to be detected. This report summarizes the preliminary results of the outbreak investigation. Aflatoxin poisoning likely will continue to be a public health problem until culturally appropriate storage methods for dry maize are implemented by the local population. In addition, enhanced surveillance for human aflatoxin poisoning and testing of commercially sold maize for aflatoxin levels will lead to long-term improvements in public health. PMID- 15343147 TI - An outbreak of norovirus gastroenteritis at a swimming club--Vermont, 2004. AB - John Snow's historic investigation of a severe epidemic of cholera traced the cause of infection to a common water source. Today, 150 years later, waterborne diseases remain a public health problem, and similar investigations are used to identify the source of infection. On February 3, 2004, the Vermont Department of Health (VDH) was notified of an outbreak of acute gastroenteritis among children whose only common exposure was attendance at a swimming club the previous weekend (January 31-February 1). This report summarizes the results of an investigation conducted by VDH and CDC, which determined the cause of the outbreak to be a combination of stool contamination, a blocked chlorine feed tube, and multiple lapses of pool-maintenance procedures. The findings underscore the importance of correct pool maintenance for rapid identification of water-quality problems to prevent outbreaks of swimming pool-associated illness. PMID- 15343148 TI - West Nile virus activity--United States, August 25-31, 2004. AB - During August 25-31, a total of 210 cases of human West Nile virus (WNV) illness were reported from 14 states (Arizona, California, Florida, Illinois, Indiana, Kansas, Maryland, Minnesota, Montana, Nevada, New Mexico, North Dakota, Oklahoma, and Pennsylvania). PMID- 15343149 TI - Early transcriptional changes of retinal and choroidal TGFbeta-2, RALDH-2, and ZENK following imposed positive and negative defocus in chickens. AB - PURPOSE: Imposing defocus to the retina results in compensatory changes of axial eye growth. It is not clear which factors initially contribute to this process and whether they act on the post-translational, translational, or transcriptional level. We have measured early changes in mRNA levels, in response to imposed negative and positive defocus, of the transcription factor ZENK, the retinoic acid synthesis enzyme RALDH-2, and the growth factor TGFbeta-2. METHODS: Chickens 11 days of age were unilaterally treated with positive or negative spectacle lenses of 7 D power. After 0, 15, 30, and 120 min, mRNA was extracted from retina and choroid, and the concentration of the mRNAs of the three candidates was measured by quantitative real time PCR in both eyes. RESULTS: ZENK in the retina and RALDH-2 in the choroid displayed parallel signs of defocus dependent changes in mRNA levels after 15 or 30 min, respectively. ZENK mRNA levels were reduced in the retina after 15 min with both types of lenses but were then up regulated at 30 min with positive lenses and down regulated with negative lenses, similar to the previously observed changes in ZENK protein levels. Changes of RALDH-2 and TGFbeta-2 mRNA levels were confined to the choroid. Treatment with negative lenses resulted in a rapid (15 min) and persistent decrease in TGFbeta-2 mRNA concentration in the choroid. Negative lenses provoked parallel but less pronounced alterations in the open fellow eyes. CONCLUSIONS: Imposed defocus triggers extensive transcriptional changes of ZENK in the retina, and of TGFbeta 2 and RALHD-2 in the choroid. Changes in retina and choroid are rapid, show no phase delay with respect to each other, and can be considered, in the case of RALDH-2 and ZENK, as specific for the sign of imposed defocus. They occur prior to any morphological changes. This is consistent with a role in causing or controlling later changes in eye growth. PMID- 15343150 TI - The ldis1 lens mutation in RIIIS/J mice maps to chromosome 8 near cadherin 1. AB - PURPOSE: We have discovered a spontaneous and severe mutation that leads to partial or complete disruption of the lens and cataract in the RIIIS/J inbred strain of mice. The purpose of this study was to determine the mode of inheritance, specificity, and range of phenotypes using histological, ophthalmic, quantitative electron microscopic, and microarray-based methods. We also have fine-mapped the mutation, ldis1 (lens disrupter 1), and have evaluated positional candidate genes. METHODS: Eyes from mutant RIIIS/J animals and from an F2 intercross between RIIIS/J and DBA/2J were examined and scored to map the ldis1 mutation. Axons in the optic nerve were counted. Messenger RNA from mutant eyes was hybridized to Affymetrix short oligomer microarrays and compared to five control strains. Expression differences were used to evaluate molecular sequellae of the mutation. RESULTS: Mice that are homozygous for ldis1 have small eyes. Lenses are without exception opaque, deformed, dislocated, fragmented, and small. In contrast, retinal architecture and ganglion cell numbers are within normal range. We have not detected any other ldis1-associated ocular or systemic abnormalities. ldis1 is recessive and maps to chromosome 8 at about 106.5 Mb between D8Mit242 and D8Mit199 with a peak LOD score near cadherin 1. The homologous human chromosomal interval is 16q22.1. The expression of several downstream crystallin transcripts are severely affected in the mutant, as are the expression levels of multiple members of the transforming growth factor superfamily and the glutathione S-transferases. CONCLUSIONS: We have discovered and mapped a recessive mutation to mouse chromosome 8 between 105 and 109 Mb. Homozygous mutant mice have a selective and severe effect on lens integrity. On the basis of the phenotype and the locus position, several candidate genes have been identified. PMID- 15343151 TI - Retention and resistance of preparations for cast restorations. 1980. PMID- 15343152 TI - The effect of adult growth on an anterior maxillary single-tooth implant: a clinical report. AB - This clinical report describes an apparent change in the position of a metal ceramic crown cemented, for approximately 1 decade, on a single anterior implant. The continuing growth of the adult facial bones is presented as a plausible explanation for this clinical scenario. PMID- 15343153 TI - Correcting a nonparallel implant abutment for a mandibular overdenture retained by two implants: a clinical report. AB - A mandibular overdenture supported by 2 implants is an effective treatment alternative for the maladaptive denture wearer. The implants may be either splinted, using a bar, or nonsplinted and retained by attachments, such as balls or magnets. When the use of a ball attachment is contemplated, the implant abutments should be parallel to one another along the path of insertion. The following report describes the procedure to correct a nonparallel implant abutment to retain a mandibular overdenture. PMID- 15343154 TI - Fixed partial denture supported by all-ceramic copings: a clinical report. AB - This clinical report describes the prosthodontic treatment of a partially edentulous patient who had a surgical closure of bilateral cleft of the lips, alveolar processes, and palate. The prosthodontic treatment included the fabrication of a "telescopic" fixed partial denture supported by reinforced all ceramic primary copings. The use of all-ceramic primary copings, rather than gold copings, offers an alternative fixed partial denture and improves the esthetic result in certain clinical situations. PMID- 15343155 TI - Combined prosthodontic and orthodontic treatment of a patient with a Class III skeletal malocclusion: a clinical report. AB - This clinical report describes a multidisciplinary approach for the treatment of a patient with Angle Class III skeletal malocclusion and decreased occlusal vertical dimension. An overlay removable partial denture (ORPD) was used to reestablish the occlusal vertical dimension (OVD). After the trial and adjustment period, the reduced lower anterior dentofacial height was orthodontically increased and the negative horizontal overlap was corrected. A maxillary precision attachment RPD and a mandibular fixed partial denture and metal ceramic crowns were fabricated to satisfy esthetic and functional requirements. PMID- 15343156 TI - Resin bonded attachments for maxillary obturator retention: a clinical report. AB - This clinical report describes the prosthetic rehabilitation of a maxillary defect with an obturator retained by extracoronal resilient attachments. Resin bonded cast-metal units were used to retain and stabilize the obturator through abutment contour modification, addition of retentive elements, and primary abutment splinting. The resulting rehabilitation conserved tooth structure and satisfied both esthetic and functional needs. PMID- 15343157 TI - Investigation into the nature of dentin resin tags: a scanning electron microscopic morphological analysis of demineralized bonded dentin. AB - STATEMENT OF PROBLEM: While the formation of the dentin/adhesive hybrid layer has been generally established, the infiltration and flow of the adhesive resin inside the acid treated dentinal tubules remains controversial. PURPOSE: The aim of the present study was to investigate and review the current interpretation of resin tags by means of scanning electron microscopic (SEM) observation. MATERIAL AND METHODS: Eight noncarious, human third molars were cut transversally and then longitudinally to obtain 8 middle-to-deep dentinal surfaces. The dentin was etched with 37% phosphoric acid (H 3 PO 4 ) gel for 10 seconds and then rinsed with water for 20 seconds. The dentin was kept moist by removing the excess water with a damp cotton pellet. The conditioned dentin was treated with a dentin bonding agent (Single Bond) and was light-polymerized for 20 seconds. A 0.2- to 0.5-mm layer of flowable composite (Tetric Flow) was then applied to the bonded dentin followed by 2 layers (2 mm each) of composite (Z 250). Each composite was light-polymerized for 40 seconds. Subsequently, the specimens were cut lengthwise into 2 halves and randomly divided into 4 groups (n=4), according to the surface preparation modality of the sectioned surface: Group EA: ethylenediamine tetraacetic acid, Group PA3: H 3 PO 4, Group PA120: H 3 PO 4 + NaOCl, and Group CA: HCl + NaOCl. Two additional teeth (Group N) were cut lengthwise into 2 halves and served as the control. The sectioned surfaces were treated with HCl and NaOCl. All specimens were processed for SEM observation. RESULTS: Specimens from Groups EA, PA3, PA120, CA, and N showed filamentous structures that were tens of microns long. Some filaments presented split-ends with hollow structures and very thin walls. Others made sharp hairpin turns indicating they were soft and compliant. CONCLUSIONS: Conventional SEM techniques, which are currently used to detect resin tags, actually identified filamentous organic structures, supposedly glycosaminoglycans, which were resistant to conventional specimen preparation techniques. The organic component showed a strong resemblance to the lamina limitans contained within the dentinal tubules. Over-reliance on SEM morphology has led to much confusion about the depth of penetration of resin tags. PMID- 15343158 TI - Influence of polymerization mode of dual-polymerized resin direct core foundation systems on bond strengths to bovine dentin. AB - STATEMENT OF PROBLEM: Little is known about how various polymerization combinations of dual-polymerized adhesives and resin core foundation pastes affect dentin bond strength. PURPOSE: This study examined the influence of different polymerization modes of dual-polymerized resin direct core foundation systems on dentin bond strength using bovine dentin. MATERIAL AND METHODS: Two commercially available dual-polymerized resin direct core foundation systems, Clearfil DC Core and UniFil Core, were studied. Bovine mandibular incisors were mounted in autopolymerizing resin and the facial dentin surfaces were ground wet on 600-grit SiC paper. Dentin surfaces were treated according to manufacturer's recommendations, and dual-polymerized adhesives were applied and dual-polymerized (control) or only chemically polymerized. The core foundation resins were condensed into the mold (4 x 2-mm) and were also either dual-polymerized or only chemically polymerized. Eighty specimens (n=10) were stored in 37 degrees C water for 24 hours, then shear tested at a crosshead speed of 1.0 mm/min in a universal testing machine. Fourier transformation infrared spectroscopy was used to measure the degree of conversion of dual-polymerized resin core foundation pastes with both modes of polymerization. One- and 2-way ANOVA, Tukey HSD tests, and Student t -tests were performed (alpha=.05). RESULTS: The highest mean bond strengths (+/ SD) were obtained when the adhesives and resin pastes were dual-polymerized for both core foundation systems (14.6 +/- 2.1 MPa for Clearfil DC Core, 15.6 +/- 2.2 MPa for UniFil Core). Chemical polymerization of dual-polymerized adhesives and resin pastes resulted in significantly lower bond strength (2.8 +/- 0.6 MPa for Clearfil DC Core, 8.1 +/- 2.2 MPa for UniFil Core) ( P <.001). The degree of conversion for the resin pastes decreased significantly ( P <.05) when only chemical polymerization was employed, from 81.5% to 76.1% for Clearfil DC Core and 86.7% to 81.2% for UniFil Core. CONCLUSION: The data suggests that the dentin bond strengths of the dual-polymerized resin direct core foundation systems are highest when both adhesive and resin core paste are dual polymerized. PMID- 15343159 TI - The effect of airborne-particle abrasion on the shear bond strength of four restorative materials to enamel and dentin. AB - STATEMENT OF PROBLEM: The role of airborne-particle abrasion on the bond strength of restorative materials to enamel and dentin is controversial. PURPOSE: The purpose of this study was to evaluate the effect of airborne-particle abrasion on the shear bond strengths of 4 restorative materials to enamel and dentin. MATERIAL AND METHODS: One hundred twelve extracted human maxillary anterior teeth were divided into 2 groups of 56 teeth each. In the first group flat enamel surfaces were prepared, and in the second flat dentin surfaces were prepared. Half the teeth in each group (n=28) were airborne-particle abraded. The remaining 28 teeth in each group were prepared with silicone carbide paper (control). Specimens in both groups were divided into 4 subgroups (n=7). A composite, a compomer, a resin-modified glass ionomer cement, and a conventional glass ionomer cement were applied to enamel and dentin surfaces (3 mm diameter and 4 mm height). After 24 hours of storage in water, specimens were subjected to shear forces in a universal testing machine at a crosshead speed of 0.5 mm/min until fracture. Shear bond strength data (MPa) were analyzed with 3-way ANOVA and Tukey HSD tests (alpha=.05). RESULTS: Airborne-particle-abraded specimens showed significantly higher shear bond strengths than control specimens. The 2-way interaction between tooth structure and restorative materials was significant. CONCLUSION: The use of airborne-particle abrasion increased the shear bond strength of restorative materials tested to enamel and dentin. PMID- 15343160 TI - Vertical marginal discrepancy of ceramic copings with different ceramic materials, finish lines, and luting agents: an in vitro evaluation. AB - STATEMENT OF PROBLEM: Prosthetic restorations that fit poorly may affect periodontal health and occlusion. Studies that have evaluated the accuracy of fit of ceramic restorations before and after cementation assessed primarily intracoronal restorations. PURPOSE: This in vitro study evaluated the effect of different finish lines, ceramic manufacturing techniques, and luting agents on the vertical discrepancy of ceramic copings. MATERIAL AND METHODS: Two stainless steel molars were prepared for complete crowns with 2 different finish lines (heavy chamfer and rounded shoulder); each molar was duplicated to fabricate 90 copings. A total of 180 copings generated 18 groups (n=10 for each finish line coping material-luting agent combination). Luting agents tested included zinc phosphate, resin-modified glass ionomer (Fuji Plus), and resin composite cements (Panavia F). A metal frame was developed on which to screw the stainless steel model and a ceramic coping; the distance (microm) between 2 predetermined points was measured before and after cementation by a profile projector under a torquing force. A 4-way ANOVA with repeated measurements was performed to assess the influence of each factor in the vertical marginal discrepancy: 3 between-coping factors (finish line-coping material-luting agent) and 1 within-coping factor (before and after cementation) (alpha=.05). RESULTS: Procera copings presented the lowest mean values ( P <.05) of vertical marginal discrepancy before and after cementation (25/44 microm) when compared to Empress 2 (68/110 microm) and InCeram Alumina copings (57/117 microm), regardless of any combinations among all finish lines and luting agents tested. CONCLUSION: Considering each factor separately, the ceramic manufacturing technique appeared to be the most important factor tested for the definitive vertical discrepancy of all-ceramic copings, with lower mean values for Procera copings. PMID- 15343161 TI - Fracture resistance of metal-free composite crowns-effects of fiber reinforcement, thermal cycling, and cementation technique. AB - STATEMENT OF PROBLEM: The improved mechanical properties of contemporary composites have resulted in their extensive use for the restoration of posterior teeth. However, the influence of fiber reinforcement, cementation technique, and physical stress on the fracture resistance of metal-free crowns is unknown. PURPOSE: This in vitro study evaluated the effect of fiber reinforcement, physical stress, and cementation methods on the fracture resistance of posterior metal-free Sinfony crowns. MATERIAL AND METHODS: Ninety-six extracted human third molars received a standardized tooth preparation: 0.5-mm chamfer preparation and occlusal reduction of 1.3 to 1.5 mm. Sinfony (nonreinforced crowns, n=48) and Sinfony-Vectris (reinforced crowns, n=48) crowns restoring original tooth contour were prepared. Twenty-four specimens of each crown type were cemented, using either glass ionomer cement (GIC) or resin cement. Thirty-two crowns (one third) were stored in humidity for 48 hours. Another third was exposed to 10,000 thermal cycles (TC) between 5 degrees C and 55 degrees C. The remaining third was treated with thermal cycling and mechanical loading (TCML), consisting of 1.2 million axial loads of 50 N. The artificial crowns were then vertically loaded with a steel sphere until failure occurred. Significant differences in fracture resistance (N) between experimental groups were assessed by nonparametric Mann Whitney U-test (alpha=.05). RESULTS: Fifty percent of the Sinfony and Sinfony Vectris crowns cemented with glass ionomer cement loosened after thermal cycling. Thermal cycling resulted in a significant reduction in the mean fracture resistance for Sinfony crowns cemented with GIC, from 2037 N to 1282 N (P=.004). Additional fatigue produced no further effects. Fiber reinforcement significantly increased fracture resistance, from 1555 N to 2326 N (P=.001). The minimal fracture resistance was above 600 N for all combinations of material, cement and loading. CONCLUSION: Fracture resistance of metal-free Sinfony crowns was significantly increased by fiber reinforcement. Adhesive cementation may be recommended to avoid cementation failure. PMID- 15343162 TI - In vitro shear bond strength of cementing agents to fixed prosthodontic restorative materials. AB - STATEMENT OF PROBLEM: Durable bonding to fixed prosthodontic restorations is desirable; however, little information is available on the strength of the bond between different cements and fixed prosthodontic restorative materials. PURPOSE: This study determined the shear-bond strength of cementing agents to high-gold content alloy castings and different dental ceramics: high-strength aluminum oxide (Procera AllCeram), leucite-reinforced (IPS Empress), and lithium disilicate glass-ceramic (IPS Empress 2). MATERIAL AND METHODS: Prepolymerized resin composite cylinders (5.5 mm internal diameter, n=20) were bonded to the pretreated surfaces of prosthodontic materials. High-gold-content alloy and high strength aluminum oxide surfaces were airborne-particle-abraded, and pressable ceramics were hydrofluoric acid-etched and silanized prior to cementing. The cementing agents tested were a zinc-phosphate cement (Fleck's zinc cement), glass ionomer cements (Fuji I, Ketac-Cem), resin-modified glass ionomer cements (Fuji Plus, Fuji Cem, RelyX Luting), resin cements (RelyX ARC, Panavia F, Variolink II, Compolute), and a self-adhesive universal resin cement (RelyX Unicem). Half the specimens (n=10) were tested after 30 minutes; the other half (n=10) were stored in distilled water at 37 degrees C for 14 days and then thermal cycled 1000 times between 5 degrees C and 55 degrees C prior to testing. Shear-bond strength tests were performed using a universal testing machine at a constant crosshead speed of 0.5 mm/min. Statistical analysis was performed by multifactorial analysis of variance taking interactions between effects into account. For multiple paired comparisons, the Tukey method was used (alpha=.05). RESULTS: In a 3-way ANOVA model, the main factors substrate, cement, time, and all corresponding interactions were statistically significant (all P <.0001). In subsequent separate 1-way or 2-way ANOVA models for each substrate type, significant differences between cement types and polymerizing modes were found (all P <.001). None of the cement types provided the highest bonding values with all substrate types. CONCLUSION: After 14 days of water storage followed by thermal cycling, only the self-adhesive universal resin cement (RelyX Unicem) and 2 of the resin cements (Panavia F and Compolute) exhibited strong bond strengths to specific prosthodontic materials. In contrast, zinc-phosphate, glass ionomer, and resin modified glass ionomer cements showed the lowest values of all tested cementing agents after 14 days of water storage followed by thermal cycling. PMID- 15343163 TI - Effect of casting technique on surface roughness and consequent mass loss after polishing of NiCr and CoCr base metal alloys: a comparative study with titanium. AB - STATEMENT OF PROBLEM: Surface roughness of cast metal frameworks may lead to difficulties in finishing or polishing procedures and weaken the framework. PURPOSE: The aim of this study was to assess the surface roughness of 2 base metal alloys, submitted to different casting techniques, to determine the influence of surface roughness on loss of mass after polishing compared to commercially pure titanium castings. MATERIAL AND METHODS: Forty disk-shaped wax patterns (8 x 2 mm) were randomly assigned to 5 groups (n=8): (1) CoCr alloy (Wironit), acetylene-oxygen flame casting (WFC), (2) NiCr alloy (Verabond II), acetylene-oxygen flame casting (VFC), (3) CoCr alloy, vacuum casting (WVC), (4) NiCr alloy, vacuum casting (VVC), and (5) commercially pure Ti alloy (Ti) melted by electric arc in argon gas atmosphere. WFC and VFC served as controls. Both NiCr and CoCr alloys were invested in phosphate-bonded investment (Termocast). Ti was invested in ammonium dihydrogen phosphate (Rematitam Plus). The wax patterns and casting techniques were standardized. All specimens were cleaned with airborne-particle abrasion (aluminum oxide). Surface roughness (Ra) was assessed using a surface-test analyzer at 3 different sites. To evaluate the loss of mass, the specimens were weighed on a precision balance, then polished with 150-600 grain sandpaper until clinically acceptable (minimum roughness of 0.09 microm). The specimens were then reweighed. A 1-way analysis of variance (ANOVA) and Tukey HSD test (alpha=.05) were performed. RESULTS: Statistical analysis showed that VVC and WVC had significantly (P=.0050; P=.0057) smoother surfaces (2.43 +/- 0.53 and 2.23 +/- 0.49, respectively) than VFC and WFC (2.99 +/- 0.44 and 2.83 +/- 0.61, respectively), but were not significantly different from Ti (2.49 +/- 0.62). The loss of mass (in %) was not significantly different for any group (3.18 +/- 0.72, 3.14 +/- 0.93, 3.36 +/- 1.05%, and 4.14 +/- 1.28% for VVC, WVC, VFC, and WFC, respectively). The mass loss of Ti was 4.32 +/- 1.16. CONCLUSION: Within the limitations of this study, the base metal alloys submitted to vacuum casting showed decreased surface roughness, similar to that of titanium, compared to base metal alloys submitted to acetylene-oxygen flame casting. There were no significant differences in loss of mass after polishing for all tested specimens. PMID- 15343164 TI - Tensile properties and hardness of cast Fe-Pt magnetic alloys. AB - STATEMENT OF PROBLEM: There is little information about the mechanical properties of castable magnetic attachments made of iron-platinum (Fe-Pt) alloys after heat treatment. MATERIAL AND METHODS: Dumbbell-shaped tensile specimens were cast with 5 alloys: 2 types of Fe-Pt alloys for magnets and keepers, pure Ti, Ti-6Al-7Nb, and Co-Cr alloy. The tensile specimens of the Fe-Pt magnets and keepers underwent different heat treatments to obtain the appropriate properties for a magnet and a keeper. Vickers hardness was measured prior to testing. Tensile testing was conducted in a universal testing machine at a crosshead speed of 2 mm/min until failure. Ultimate tensile strength, yield strength, elongation, and modulus of elasticity were recorded. The data were analyzed using 1-way analysis of variance and the Tukey test (alpha=.05), fractured surfaces were examined using a scanning electron microscope (SEM). RESULTS: The highest tensile and yield strengths were found for Ti-6Al-7Nb, followed by Co-Cr, CP Ti, and the Fe-Pt keeper. There was no significant difference in ultimate tensile strength between the CP Ti and the Fe-Pt keeper. The lowest elongation was found for the Fe-Pt magnet (2.8%), whereas the Fe-Pt keeper had the highest elongation (14.0%). The Fe-Pt magnet and keeper had similar modulus of elasticity values. The hardness of the cast Fe-Pt magnets was the highest among the metals tested. SEM micrographs of the Fe-Pt keepers showed a microstructure with ductile dimple fractures, whereas the Fe-Pt magnets were characterized by brittle fractures at the grain boundaries. CONCLUSIONS: The mechanical properties of the cast Fe-Pt keepers were similar to those of the cast pure Ti. The Fe-Pt magnets were extremely hard and brittle. PMID- 15343165 TI - Effect of denture base surface pretreatment on microleakage of a silicone-based resilient liner. AB - STATEMENT OF PROBLEM: Microleakage between resilient liner and denture base resins is a significant clinical problem, often responsible for debonding of the resilient liner from the denture base resin. PURPOSE: This study investigated the effect of 2 surface treatments, airborne-particle abrasion (APA) and wetting with methyl methacrylate monomer (MMA), on microleakage between a silicone-based resilient liner and denture base resin using a gamma camera imaging technique. MATERIAL AND METHODS: Thirty-three specimens, each having 2 plates measuring 40 x 40 x 2 mm, were prepared by packing and processing an acrylic denture base resin (QC-20) into square plates following manufacturer's instructions. Specimens were divided into 3 groups (n=11) as APA-, MMA-, and control-treatment groups. For the APA group, the inner surfaces of both plates were airborne-particle abraded with 250-microm Al 2 O 3 particles and, for the MMA group, surfaces were treated with monomer (QC-20). Control specimens were not surface treated. Following application of an adhesive (Ufi Gel P-specific), a silicone lining material (Ufi Gel P) was prepared and applied to the inner surfaces of all 33 specimens. Eleven size-matched polymethyl methacrylate (PMMA) specimen blocks (40 x 40 x 6 mm) were prepared to calculate the level of residual radioactivity for the denture base itself, the entire outer surface count (OSC). All specimens and PMMA blocks were immersed in a radioactive solution (thallium-201 chloride) for 24 hours. Specimen activities (gamma-ray cts/sec, representing thallium-201 concentration) were then measured using a high-resolution gamma camera. The amount of OSC-subtracted total specimen counts was a direct indicator of the quantity of inward diffusing tracer. The subtracted values were analyzed using a 1-way analysis of variance (ANOVA) and Bonferroni multiple comparison tests (alpha=.05). RESULTS: OSC levels averaged 754 +/- 110 gamma-ray cts/sec. OSC-subtracted APA, control, and MMA values were 5,546 +/- 1,534, 3,392 +/- 738, and 1,405 +/- 392 gamma-ray cts/sec, respectively. All 3 groups were significantly different ( P <.05) from each other. Surface wetting with MMA showed the lowest microleakage values among all specimen groups. CONCLUSION: In terms of microleakage, surface treatment with MMA monomer preceding the adhesive application demonstrated lower values than adhesive application alone. APA pretreatment resulted in 4 times the microleakage found in MMA-treated specimens, and 1.5 times the microleakage of the untreated control. None of the surface pretreatments completely prevented microleakage. Microleakage between the silicone-based resilient liner and denture base resin can be quantitatively determined using the gamma camera imaging technique. PMID- 15343166 TI - Evaluation of varying amounts of thermal cycling on bond strength and permanent deformation of two resilient denture liners. AB - STATEMENT OF PROBLEM: Two problems found in prostheses with resilient liners are bond failure to the acrylic resin base and increased permanent deformation due to material aging. PURPOSE: This in vitro study evaluated the effect of varying amounts of thermal cycling on bond strength and permanent deformation of 2 resilient denture liners bonded to an acrylic resin base. MATERIAL AND METHODS: Plasticized acrylic resin (PermaSoft) or silicone (Softliner) resilient lining materials were processed to a heat-polymerized acrylic resin (QC-20). One hundred rectangular specimens (10 x 10-mm 2 cross-sectional area) and 100 cylindrically shaped specimens (12.7-mm diameter x 19.0-mm height) for each liner/resin combination were used for the tensile and deformation tests, respectively. Specimen shape and liner thickness were standardized. Specimens were divided into 9 test groups (n=10) and were thermal cycled for 200, 500, 1000, 1500, 2000, 2500, 3000, 3500, and 4000 cycles. Control specimens (n=10) were stored for 24 hours in water at 37 degrees C. Mean bond strength, expressed as stress at failure (MPa), was determined with a tensile test using a universal testing machine at a crosshead speed of 5 mm/min. Analysis of failure mode, expressed as a percent (%), was recorded as either cohesive, adhesive, or both, after observation. Permanent deformation, expressed as a percent (%), was determined using ADA specification no. 18. Data from both tests were examined with a 2-way analysis of variance and a Tukey test (alpha=.05). RESULTS: For the tensile test, Softliner specimens submitted to different thermal cycling regimens demonstrated no significantly different bond strength values from the control; however, there was a significant difference between the PermaSoft control group (0.47 +/- 0.09 MPa [mean +/- SD]) and the 500 cycle group (0.46 +/- 0.07 MPa) compared to the 4000 cycle group (0.70 +/- 0.20 MPa) ( P <.05). With regard to failure type, the Softliner groups presented adhesive failure (100%) regardless of specimen treatment. PermaSoft groups presented adhesive (53%), cohesive (12%), or a combined mode of failure (35%). For the deformation test, there was no significant difference among the Softliner specimens. However, a significant difference was observed between control and PermaSoft specimens after 1500 or more cycles (1.88% +/- 0.24%) ( P <.05). CONCLUSIONS: This in vitro study indicated that bond strength and permanent deformation of the 2 resilient denture liners tested varied according to their chemical composition. PMID- 15343167 TI - Effect of multi-adhesive layering on retention of extraoral maxillofacial silicone prostheses in vivo. AB - STATEMENT OF PROBLEM: Loss of retention of maxillofacial prostheses often makes the margin visible or the prosthesis dislodge. Using several medical adhesives in combination may improve retention. PURPOSE: The purpose of this study was to investigate the effect of single- and multi-adhesive layering of 2 adhesives on the retention of maxillofacial silicone elastomer strips adhered to the skin of human forearms using a peel test. MATERIAL AND METHODS: Power analysis from a previous study and a pilot trial specified at least 20 subjects. Eight Silastic Adhesive A/MDX4-4210 silicone rubber strips (N=240) were applied in a predetermined random order to the left and right ventral forearms of 30 IRB approved human subjects. Skin-Prep Protective Dressing was applied. Secure 2 Medical Adhesive (SMA) and Epithane-3 (E3) adhesive were used alone or as SMA/E3 or E3/SMA sandwiches (from skin to prosthesis) to adhere strips. Strips were peeled 6 hours later in a universal testing machine at 10 cm/min and data reported in N/m. Paired t tests were used to evaluate left and right arm differences. A Friedman test for nonparametric correlated data with within subject design was performed, determining differences between both adhesives singly and in combination (alpha=.05). RESULTS: Tests of left-right differences were insignificant ( P =0.43), so the data from both arms were combined. Many strips with E3 did not adhere before testing and were counted as 0 adhesion. Median peel strengths (and 25th and 75th percentiles) in N/m were: SMA = 76.1 (47.1-107), E3 = 6.75 (0.0-25.9), SMA/E3 = 107 (78.0-132), and E3/SMA= 19.6 (6.99 42.4). All 4 variables were significantly different ( P <.0005). CONCLUSION: The multi-adhesive combination of SMA/E3 had the highest adhesion, followed, in order, by SMA alone, E3/ SMA, and E3 alone. Both E3 groups left a difficult-to remove residue on the skin. SMA/E3 left a halo-like residue on the skin at the periphery of the strips from the E3 leaking around the SMA. SMA remained adherent to the prosthetic material. PMID- 15343168 TI - A selective-pressure impression technique for the edentulous maxilla. AB - This article describes a selective-pressure impression technique for the edentulous maxilla that is intended to compensate for the polymerization shrinkage of heat-polymerized polymethyl methacrylate resin and provides improved palatal adaptation of the definitive denture base. PMID- 15343169 TI - Using silicone occlusal registration material as a guide for tooth preparation. PMID- 15343170 TI - A method for preventing aspiration or ingestion of fixed restorations. PMID- 15343171 TI - Academic leadership: reflections in the key of C. PMID- 15343172 TI - Novel oligosaccharides in human milk: understanding mechanisms may lead to better prevention of enteric and other infections. PMID- 15343173 TI - Assessment of maldigestion in cystic fibrosis. PMID- 15343174 TI - Expanding the role of hydroxyurea in children with sickle cell disease. PMID- 15343175 TI - Tips for the use of antihypertensive drugs: DELTAREPROSI. PMID- 15343176 TI - Alcohol research in the emergency department setting: methodological challenges and real-world implications. PMID- 15343177 TI - Ethical issues in newborn screening research: lessons from the Wisconsin cystic fibrosis trial. PMID- 15343178 TI - Human milk oligosaccharides are associated with protection against diarrhea in breast-fed infants. AB - OBJECTIVE: To determine the association between maternal milk levels of 2-linked fucosylated oligosaccharide and prevention of diarrhea as a result of Campylobacter, caliciviruses, and diarrhea of all causes in breast-fed infants. STUDY DESIGN: Data and banked samples were analyzed from 93 breast-feeding mother infant pairs who were prospectively studied during 1988-1991 from birth to 2 years with infant feeding and diarrhea data collected weekly; diarrhea was diagnosed by a study physician. Milk samples obtained 1 to 5 weeks postpartum were analyzed for oligosaccharide content. Data were analyzed by Poisson regression. RESULTS: Total 2-linked fucosyloligosaccharide in maternal milk ranged from 0.8 to 20.8 mmol/L (50%-92% of milk oligosaccharide). Moderate-to severe diarrhea of all causes (n=77 cases) occurred less often (P=.001) in infants whose milk contained high levels of total 2-linked fucosyloligosaccharide as a percent of milk oligosaccharide. Campylobacter diarrhea (n=31 cases) occurred less often (P=.004) in infants whose mother's milk contained high levels of 2'-FL, a specific 2-linked fucosyloligosaccharide, and calicivirus diarrhea (n=16 cases) occurred less often (P=.012) in infants whose mother's milk contained high levels of lacto-N-difucohexaose (LDFH-I), another 2-linked fucosyloligosaccharide. CONCLUSION: This study provides novel evidence suggesting that human milk oligosaccharides are clinically relevant to protection against infant diarrhea. PMID- 15343179 TI - Survey of vitamin A supplementation for extremely-low-birth-weight infants: is clinical practice consistent with the evidence? AB - OBJECTIVE: To survey the attitudes and practices among level III neonatal intensive care units in the United States regarding vitamin A supplementation for extremely-low-birth-weight (ELBW; birth weight < or =1000 g) infants. Study design A pretested questionnaire regarding vitamin A supplementation was distributed to all (n=102) neonatal-perinatal training program directors (TPD) and 105 randomly selected directors of level III neonatal intensive care units (nontraining program directors, NTPD). RESULTS: Ninety-nine percent of TPD and 94% of NTPD responded. In a minority of programs (20% TPD, 13% NTPD), >90% of eligible extremely-low-birth-weight neonates are supplemented with vitamin A, whereas in most programs (69% TPD, 82% NTPD), routine supplementation is not practiced. Most centers (91% TPD, 81% NTPD) supplementing vitamin A use a dose of 5000 IU IM 3 times per week for 4 weeks. The most common reason that TPD give for not supplementing vitamin A is the perceived small benefit, whereas the most common reason for NTPD is that they consider the intervention unproven. CONCLUSIONS: These findings indicate inconsistency in practicing evidence-based medicine in neonatal practice, where therapies are often administered on the basis of weaker evidence of safety and benefit than supports vitamin A supplementation. Educational interventions may be required to endorse the benefits and safety of vitamin A supplementation. PMID- 15343180 TI - Use of nonsterile instruments for examination for retinopathy of prematurity in the neonatal intensive care unit. AB - OBJECTIVE: Use of nonsterile eyelid speculae and scleral depressors during screening examinations for retinopathy of prematurity (ROP) in the neonatal intensive care unit (NICU) violates existing infection control recommendations. The objective of this study was to characterize current instrument management and to estimate the prevalence of use of nonsterile instruments. STUDY DESIGN: NICU nurse managers were surveyed. Questionnaires were mailed to 735 NICUs from The United States Neonatologists and Perinatologists Directory. RESULTS: Of 300 surveys returned, 290 reported ROP examinations in their NICUs. Two hundred ten (72%) of 290 NICUs reported supplying instruments. Seventy-five (36%) of 210 NICUs supplying instruments reported a best practice on the basis of current guidelines. Twenty-six (9%) of 290 NICUs reported eye infections as a result of ROP examinations. Reported infection was more frequent (19%) among NICUs reporting reuse of instruments ( P =.019) and more frequent (17%) among NICUs in which ophthalmologists brought their own instruments ( P =.021). Only one (2%) NICU reporting best practice also reported eye infections ( P =.004). CONCLUSIONS: ROP instrument management is inconsistent within and between NICUs. Without the exclusive use of sterile instruments, an increased risk of nosocomial infection might exist. Greater attention to sterile instrument management is recommended. PMID- 15343181 TI - Is surfactant therapy beneficial in the treatment of the term newborn infant with congenital diaphragmatic hernia? AB - OBJECTIVES: To determine the impact of surfactant replacement on survival, need for extracorporeal membrane oxygenation (ECMO), and chronic lung disease in term infants with prenatally diagnosed congenital diaphragmatic hernia (CDH). STUDY DESIGN: Prenatally diagnosed infants born at > or =37 weeks' gestation with immediate distress at delivery and no other major congenital anomalies, who were enrolled in the CDH Registry, were analyzed. For univariate analysis, chi 2 tests were used for categoric variables and unpaired t tests for nominal variables. Multiple logistic regression was used to calculate adjusted odds ratios. RESULTS: Eligible infants (n = 522) were identified. Demographic variables were similar between the surfactant-treated (n = 192) and nonsurfactant-treated (n = 330) groups, with the exception of race (white, 88.0% vs 71.2%; P =.0007). The use of ECMO and incidence of chronic lung disease were higher (59.8 vs 50.6, P =.04; 59.9 vs 47.6, P =.0066) and survival lower in the surfactant-treated cohort (57.3 vs 70.0, P =.0033). Adjusted logistic regression for use of ECMO, survival, and chronic lung disease resulted in odds ratios inconsistent with an improved outcome associated with surfactant use. CONCLUSIONS: This analysis shows no benefit associated with surfactant therapy for term infants with a prenatal diagnosis of isolated CDH. PMID- 15343182 TI - Ursodeoxycholic acid (UDCA) therapy in very-low-birth-weight infants with parenteral nutrition-associated cholestasis. AB - OBJECTIVE: To determine the effect of ursodeoxycholic acid (UDCA) in very-low birth-weight (VLBW) infants with parenteral nutrition-associated cholestasis (PNAC). STUDY DESIGN: A retrospective study of all VLBW infants with PNAC who were admitted to a tertiary referral center was conducted. Patients were classified as treatment group (receiving UDCA within 14 days after onset of cholestasis) or control group (no medical treatment). Patients who received abdominal surgery were excluded. RESULTS: A total of 30 patients were recruited, including 12 in the treatment group and 18 in the control group. The demographic data, total fasting duration, onset of cholestasis, age to tolerance of full feeds, and the duration of parenteral nutrition (PN) before the onset of cholestasis were comparable between the two groups. There was a trend in the control group to later onset of cholestasis. The patients who received UDCA therapy with doses of 10 to 30 mg/kg/day had a shorter duration of cholestasis than the control group (62.8 vs 92.4 days, P=.006). Furthermore, the peak serum levels of direct bilirubin also was significantly lower in the treatment group. CONCLUSION: UDCA can improve the course of PNAC in VLBW infants. PMID- 15343184 TI - Use of fecal elastase-1 to classify pancreatic status in patients with cystic fibrosis. AB - OBJECTIVE: To test the hypothesis that some patients with cystic fibrosis (CF) are misclassified as pancreatic insufficient, using fecal elastase-1 (FE-1) to define pancreatic status. STUDY DESIGN: Subjects with CF at 33 CF centers filled out questionnaires and submitted a stool specimen that was analyzed for FE-1. Subjects taking pancreatic enzyme supplements (PES) were asked to discontinue them and perform a 3-day fecal fat balance study if their FE-1 was >200 microg/g stool and they had never had pancreatitis. RESULTS: The median value for FE-1 in 1215 subjects was 0 microg/g stool (range, 0-867). There was a significant difference between patients who had been prescribed PES (n=1131) and those who had FE-1 <200 microg/g stool (n=1074; P<.0001). Sixty-seven subjects met criteria for discontinuation of PES. The mean coefficient of fat absorption for these subjects was 96.1%. CONCLUSIONS: FE-1 is an accurate, easily obtained screening test to classify pancreatic status in patients with CF. This information is important for prognostication, treatment, and to avoid misclassification in clinical research. Measurement of FE-1 should become a standard of care for patients with CF. PMID- 15343185 TI - Cystic fibrosis-associated liver disease: a population-based study. AB - OBJECTIVES: The aim of this study was to explore the clinical factors associated with the development of cystic fibrosis-associated liver disease (CFALD). STUDY DESIGN: This was a case-control study of all children (age 5-18 years) with established CFALD in the Republic of Ireland between January 1999 and June 2000. Each child was pair matched for age and sex with a patient with cystic fibrosis (CF) without evidence of liver disease. Only children with clinically overt liver disease were enrolled in the disease group. RESULTS: Patients with established CFALD (n = 42; 26 boys) were enrolled. Children with CFALD had worse forced expiratory volume in 1 second values than those without CFALD. However, chest radiography and clinical scores did not differ between groups. Height (mean difference, -4.2 cm [95% confidence interval [CI], -7.41 to -0.90], P =.014), weight (mean difference, -3.21 kg [95% CI, -6.03 to -0.40], P =.026), and mid upper arm circumference (mean difference, -1.23 cm [95% CI, -2.35 to -0.12], P =.031) were significantly lower among children with CFALD. Children with CFALD were given diagnoses of CF later than children without liver disease. There were more children with meconium ileus in the control group (14 vs 4) than among those with CFALD. CONCLUSIONS: Children with established CFALD have impaired growth and nutrition, altered body composition, and worse forced expiratory volume in 1 second values. CFALD is associated with later age of diagnosis of CF. PMID- 15343186 TI - Liver disease in children with primary immunodeficiencies. AB - OBJECTIVE: To investigate clinical features and to establish optimal management in children with primary immunodeficiency (PID) and liver disease. Study design A retrospective analysis of medical records of 147 children with PID who presented with abnormal liver tests to a tertiary center. RESULTS: Clinical evidence of liver disease was documented in 35 (23.8%) patients. Of these, 22 (63%) had hepatomegaly and 14 (40%) had splenomegaly. Sclerosing cholangitis (SC) was diagnosed in 21 children (60%), based on radiological and histological criteria; 4 patients with SC on cholangiography had no biliary changes in the liver biopsy. Ultrasonography demonstrated a dilated biliary system in 14 (67%) children with SC. Of 27 children investigated for Cryptosporidium parvum (CSP), 12 (44%) were positive, including 9 of 12 with SC. Overall, 7 (20%) patients died, including 3 boys with disseminated recurrent CSP infection after successful liver transplantation (LT). Temporary deterioration of liver injury was observed in 2 CSP-positive boys with CD40 ligand deficiency (CD40LD) who were undergoing nonmyeloablative hematopoietic stem cell transplantation (HSCT). Successive liver and HSCT was curative in 1 patient with CD40LD and end-stage liver disease. CONCLUSION: SC is the most common hepatic complication of PID. Mild liver involvement could be arrested by early nonmyeloablative HSCT, whereas advanced disease may warrant combined liver and HSCT. PMID- 15343187 TI - Interferon treatment for chronic hepatitis B: enhanced response in children 5 years old or younger. AB - OBJECTIVE: To test the hypothesis that there is an improved response to interferon in children with chronic hepatitis B (HBV) who are < or =5 years of age. STUDY DESIGN: Retrospective chart review of 22 consecutive children with chronic HBV (ages 17 months to 17 years; median, 83.9 months; 14 male, 8 female) treated with interferon-alpha2b. RESULTS: Ten patients (48%) responded to treatment [HBeAg (-), Anti-HBe (+), HBV DNA (-), HBsAg (+) and normal alanine aminotransferase/aspartate aminotransferase (ALT/AST) at 6 months after treatment], and 5 seroconverted HBsAg [above plus HBsAg negative and anti-HBs (+)]. Seven of 9 patients (78%) < or =5 years of age responded (5 cleared HBsAg). Three of 13 patients (23%) >5 years of age responded. Patient age at treatment was significantly lower in responders (63 +/- 70 months) versus nonresponders (104 +/- 55 months, P =.005). AST, ALT, and HBV DNA at the start of treatment were not different between responders and nonresponders or between patients < or =5 and >5 years old. CONCLUSIONS: Interferon treatment may be more effective in younger children with chronic hepatitis B. PMID- 15343189 TI - Prevention of secondary stroke and resolution of transfusional iron overload in children with sickle cell anemia using hydroxyurea and phlebotomy. AB - OBJECTIVE: Transfusions prevent secondary stroke in children with sickle cell anemia (SCA) but also cause iron overload. Alternatives for stroke prophylaxis with effective therapy to reduce iron burden are needed. STUDY DESIGN: For 35 children with SCA and stroke, transfusions were prospectively discontinued. Hydroxyurea was prescribed for stroke prophylaxis, and phlebotomy removed excess iron. Initial patients discontinued transfusions before hydroxyurea therapy, but later patients overlapped transfusions with hydroxyurea until tolerating full dose therapy. RESULTS: Children received hydroxyurea for 42 +/- 30 months (range, 3-104 months). Hydroxyurea (26.7 +/- 4.8 mg/kg per day) led to mild neutropenia (3.9 +/- 2.3 x 10(9)/L) with significant increases in hemoglobin concentration, mean corpuscular volume, and fetal hemoglobin. Stroke recurrence rate was 5.7 events per 100 patient-years, but children receiving overlapping hydroxyurea therapy had only 3.6 events per 100 patient-years. For 26 children with >6 months of phlebotomy, 14,311 +/- 12,459 mL blood (315 +/- 214 mL/kg) was removed, with serum ferritin decreasing from a median of 2722 to 298 ng/mL. Among patients completing phlebotomy, liver biopsy documented normal histology and no excess iron deposition. CONCLUSIONS: For children with SCA and stroke, hydroxyurea effectively prevents secondary stroke and serial phlebotomy leads to complete resolution of transfusional iron overload. PMID- 15343191 TI - A randomized, placebo-controlled trial of amlodipine in children with hypertension. AB - OBJECTIVES: Evaluation of the efficacy and safety of amlodipine in hypertensive children. STUDY DESIGN: A randomized, double blinded, placebo-controlled, parallel-group, dose-ranging study was conducted at 49 centers in North and South America. The primary end point was the effect of amlodipine on systolic blood pressure (BP); secondary end points included the effect of amlodipine on diastolic BP, the effect of amlodipine as a function of dose and body size, and evaluation of safety. RESULTS: We enrolled 268 hypertensive children (mean age, 12.1 +/- 3.3 years); 84 (31.3%) had primary hypertension, and 177 (66%) were boys. Amlodipine produced significantly greater reductions in systolic BP than placebo; these were -6.9 mm Hg for 2.5 mg daily (P=.045 vs placebo) and -8.7 mm Hg for 5 mg daily (P=.005 vs placebo). The underlying cause of hypertension had no effect on the response to amlodipine. There was a significant dose-response effect of amlodipine on both systolic and diastolic BP beginning at doses > or =0.06 mg/kg per day. Systolic BP < or =95(th) percentile was achieved in 34.6% of subjects with systolic hypertension. Amlodipine was well tolerated, with just 6 children withdrawn from treatment because of drug-related adverse events. CONCLUSIONS: Amlodipine effectively lowers systolic BP in a dose-dependent manner in hypertensive children who require drug treatment. PMID- 15343192 TI - Predicting bicycle helmet stage-of-change among middle school, high school, and college cyclists from demographic, cognitive, and motivational variables. AB - OBJECTIVES: To apply Prochaska's Transtheoretical model of behavior change to bicycle helmet use among middle school, high school, and college students. STUDY DESIGN: A battery of questionnaires was administered to cyclists in the seventh and ninth grades and to college students in Phoenix, Arizona (N=797). The battery included: (1) a question to determine respondent's stage of behavior change in Prochaska's Transtheoretical model; (2) items assessing the perceived pros and cons of helmet use; (3) a bicycle safety knowledge test; and (4) demographic information. RESULTS: Forty-three percent of the students were in "Precontemplation," 17% were in either "Contemplation" or "Preparation," 16% were in either "Action" or "Maintenance," and 24% were in the "Relapse" stage of change. Grade, Sex, Knowledge, Pros, and Cons, and the Grade by sex and the Grade by knowledge interactions were significant predictors of helmet use stages. Compared with students in Precontemplation, students in the Contemplation stage were disproportionately younger and had higher Pro scores, lower Con scores, and more knowledge (except in the ninth grade). CONCLUSIONS: The Transtheoretical model of behavior change is a viable theoretical framework for designing interventions aimed at increasing bicycle helmet use in children and adolescents. PMID- 15343193 TI - Neuropsychological outcome of infant heart transplant recipients. AB - OBJECTIVE: To assess long-term neuropsychological outcomes of infant heart transplant recipients. STUDY DESIGN: Fifty-five children (mean age, 6.4 years) with a primary diagnosis of hypoplastic left heart syndrome or other cardiac disorders participated in the study. Six areas of risk were assessed (birth; preoperative, intraoperative, and postoperative status; long-term medical complications; and socioeconomic factors). Neuropsychological evaluations included the following: Wechsler Scales of Intelligence, Wechsler Individual Achievement Test, Berry Visual Motor Integration Scale, Comprehensive Evaluation of Language Function, and Achenbach Child Behavioral Checklist. RESULTS: Intellectual functioning scores revealed a mean full-scale IQ of 81 +/- 19. Intraoperative and postoperative variables (bypass time, hospital course, and lowest posttransplant pH), long-term medical complications (serious infections and posttransplant surgical procedures), and socioeconomic factors were associated with cognitive functioning. Visual-motor functioning was also related to bypass and total support time. CONCLUSIONS: Long-term outcomes reveal low average neuropsychological functioning and are related to intrasurgical postsurgical, long-term medical, and socioeconomic factors. The neuropsychological and psychosocial functioning of infant heart transplant recipients will be important to address as they mature during adolescence and adulthood. PMID- 15343194 TI - Serious illness in childhood: the different threats of cancer and diabetes from a parent perspective. AB - OBJECTIVES: To compare the incidence of disease-related distress symptoms in parents of children with cancer and diabetes. STUDY DESIGN: A total of 675 parents of patients with cancer, patients with diabetes, and control subjects were assessed for 11 distress symptom clusters. Patient and control parent mean differences were tested by 2-tailed t tests; illness groups were compared by means of analysis of variance. Distress variations as a function of time since diagnosis were examined by regression analysis. RESULTS: The distress levels of patient parents exceeded those of control parents for global distress ( P <.0001) and for most symptom subcategories. Distress levels of parents of patients with cancer (CP) significantly exceeded those of parents of patients with diabetes (DP) in anxiety ( P <.0001), physical and psychologic distress ( P <.0001), depression ( P <.005), and loneliness ( P <.05). Levels in DP matched those of CP in uncertainty, loss of control/the patient, self-esteem, disease-related fear, and sleep disturbances. Distress levels were lower in CP most distant in time from diagnosis, whereas DP showed a reversed trend. CONCLUSIONS: Parental distress patterns in childhood illness depend on illness type and time passed since diagnosis. Symptom profiles verify the need for psychosocial attention at the initial shock after the cancer diagnosis and indicate long-term consequences for many parents. In pediatric diabetes, the persistence or intensification of distress over time is of specific clinical relevance. PMID- 15343195 TI - A randomized, controlled study of insulin pump therapy in diabetic preschoolers. AB - OBJECTIVE: To compare glycemic control, safety, and parental satisfaction in preschool-aged diabetic children randomized to treatment either with continuous subcutaneous insulin infusion (CSII) or intensive insulin injection therapy. STUDY DESIGN: This clinical trial enrolled 42 patients <5 years of age who had been diagnosed with diabetes for at least 12 months. Children were randomly assigned to CSII (n = 21) or intensive insulin injection therapy (n = 21). Hemoglobin A1c (HbA1c) level was measured at baseline, 3, and 6 months. Secondary outcomes included severe hypoglycemic events, meter-detected hypoglycemia, blood sugar variability, body mass index (BMI), and satisfaction with therapy. RESULTS: Thirty-seven patients completed 6 months of therapy. There was a significant decrease in HbA1c during the study period for both groups (from 8.9% +/- 0.6% to 8.6% +/- 0.6% at 3- and 6-month visits). At 3 months, children using pumps had a significantly lower HbA1c than the injection group (8.4% vs 8.8%); however, by 6 months the two groups were similar (8.5% vs 8.7%). No differences in pre-meal blood sugar variabilities were seen between groups. Children on pumps had increases in the number of meter-detected episodes of hypoglycemia. Pump therapy was safe and well tolerated. No episodes of ketoacidosis occurred in either group, whereas one hypoglycemic seizure occurred in each group. Parents reported satisfaction with CSII, with 95% of families continuing on CSII beyond the 6 month study period. CONCLUSION: Pump therapy in preschool-aged children was not associated with clinically significant differences in glycemic control as compared with intensive injection therapy. The rationale for initiating CSII in this age group should be based on patient selection and lifestyle preference. PMID- 15343196 TI - CD25+CD4+ regulatory T cells in patients with Kawasaki disease. AB - OBJECTIVE: To investigate whether the CD25 + CD4 + regulatory T-cell population, which plays important roles not only in maintaining immunologic self-tolerance but also in controlling the magnitude and character of antimicrobial immune responses, is related to the pathophysiology of Kawasaki disease (KD). STUDY DESIGN: The patient group consisted of 54 patients (median age, 30 months; 27 female and 27 male patients) fulfilling the criteria for KD. Age-matched control subjects included 17 patients with active infections and 24 healthy children. We analyzed CD25 + CD4 + cells and the mRNA expression of Foxp3, cytotoxic T lymphocyte-associated antigen 4 (CTLA4), glucocorticoid-induced tumor necrosis factor receptor (GITR), and transforming growth factor beta in peripheral blood mononuclear cells and purified CD4 + T cells. RESULTS: The proportions of CD25 + CD4 + cells in patients with acute-phase KD (median, 2.35% of total lymphocytes) were significantly lower than those in healthy control subjects (median, 3.14%) and control subjects with disease (median, 3.15%). The proportions returned to the normal level after intravenous gammaglobulin treatment (median, 3.86%). The mRNA expression of Foxp3, CTLA4, and GITR showed similar tendencies. CONCLUSIONS: The decrease of CD25 + CD4 + regulatory T cells in the acute phase might have a role in the development of KD. PMID- 15343197 TI - Repetitive arm and hand movements (complex motor stereotypies) in children. AB - OBJECTIVE: To characterize clinical features, associated problems, and outcomes for children with complex motor stereotypies who do not have mental retardation or pervasive developmental disorders. STUDY DESIGN: We performed a record review for 40 children (63% male) aged 9 months to 17 years with complex motor stereotypies between 1993 and 2003. RESULTS: Age at onset was at or before 3 years in 90% of the sample. Symptoms occurred at least once daily in 90%. Excitement was identified as a trigger in 70%. Movements stopped when cued in 98%, and none had stereotypies during sleep. A total of 25% had comorbid attention deficit hyperactivity disorder (ADHD), and 20% had a learning disability. Family history of stereotypies was identified in 25%, tics in 33%, ADHD in 10%, and mood-anxiety disorder in 38%. Pharmacotherapy to target associated conditions was used in 40%, and behavioral therapy was used in 23%. A total of 53% identified symptoms for more than 5 years. Movements resolved in 5% of the children, improved in 33%, were unchanged in 50%, and worsened in 13%. CONCLUSIONS: The clinical course of complex motor stereotypies appears chronic. Better understanding of the clinical features of complex stereotypies in primary care settings is essential for early diagnosis and management. PMID- 15343198 TI - A randomized clinical trial of a brief motivational intervention for alcohol positive adolescents treated in an emergency department. AB - We tested whether a brief motivational interview (MI) would reduce alcohol related consequences and use among adolescents treated in an emergency department (ED) after an alcohol-related event. Patients aged 13 to 17 years (N = 152) with a positive blood alcohol concentration (BAC) by lab test or self-report were recruited in the ED and randomly assigned to receive either MI or standard care (SC). Both conditions resulted in reduced quantity of drinking during the 12 month follow-up, whereas alcohol-related negative consequences were relatively low and stayed low at follow-up. Adolescents who screened positive for problematic alcohol use at baseline reported significantly more improvement on 2 of 3 alcohol use outcomes (average number of drinking days per month and frequency of high-volume drinking) if they received MI compared with SC. We conclude that brief interventions are recommended for adolescents who present to an ED with an alcohol-related event and report preexisting problematic alcohol use. PMID- 15343199 TI - Maintaining the nelfinavir trough concentration above 0.8 mg/L improves virologic response in HIV-1-infected children. AB - Differences in virologic response were compared in 32 HIV-infected children with a nelfinavir trough concentration either below (n=7) or above (n=25) 0.8 mg/L. Virologic response at week 48 was observed in 29% of children with subtherapeutic nelfinavir troughs versus 80% in children with therapeutic nelfinavir troughs (P=.02). PMID- 15343200 TI - Monozygotic twins with discordant karyotypes: a case report. AB - A monochorionic twin pregnancy had normal male karyotype on chorionic villous sampling. At delivery, one twin presented as morphologically normal, the other as trisomy 21. A twinning event and chromosome division error shortly after conception resulted in monozygotic twins with discordant tissue karyotypes and blood chromosome chimerism for trisomy 21. PMID- 15343201 TI - Incidence of spinal cord lesions in patients with intractable constipation. AB - Abstract Spinal magnetic resonance imaging of children with intractable constipation was reviewed. Nine percent of patients had spinal cord abnormalities. Tethered cord was the most common lesion in 75% of patients. Magnetic resonance imaging is a useful tool to evaluate children with intractable constipation. PMID- 15343202 TI - Hemolytic uremic syndrome in a child with laboratory-acquired Escherichia coli O157:H7. AB - A 6-year-old girl touched an agar plate containing Escherichia coli O157:H7 while visiting a hospital laboratory, and subsequently, colitis and hemolytic uremic syndrome developed. Pulsed-field gel electrophoresis patterns of the isolate cultured from her stool and that from the laboratory were identical. PMID- 15343203 TI - Congenital segmental dilatation of the small bowel. PMID- 15343205 TI - Management and outcomes of care of fever in early infancy. PMID- 15343204 TI - Effectiveness over time of varicella vaccine. PMID- 15343206 TI - Diagnosis and testing in bronchiolitis: a systematic review. PMID- 15343207 TI - Otitis media and speech and language: a meta-analysis of prospective studies. PMID- 15343208 TI - Topiramate for migraine prevention: a randomized controlled trial. PMID- 15343211 TI - Resident attrition: is gender a factor? AB - OBJECTIVE: This study was undertaken to identify the causes of resident attrition from obstetrics and gynecology on the basis of gender. STUDY DESIGN: Two sequential questionnaires were sent to the 246 obstetrics and gynecology residency programs in the United States in March 2001 and May 2002. The programs were asked for the number of residents who left from 1997 to 2001, the postgraduate years, and genders of those residents, the reasons for leaving, and program demographics. Statistical analysis was performed with chi 2 and odds ratio calculations. RESULTS: The rate of attrition was 3% over 4 years. Female residents were 2.5 to 5 times as likely as male residents to leave because of family issues, specifically for reasons related to spouses ( P =.002). Female residents were one quarter as likely to leave to change specialty (P =.002), whereas this was the principal reason of more than half of the male residents who left. CONCLUSION: Important gender differences do exist in the reasons for attrition from obstetrics and gynecology residency programs. PMID- 15343212 TI - Cohort study of forceps delivery and the risk of epilepsy in adulthood. AB - OBJECTIVE: The purpose of this study was to investigate the relationship between forceps delivery and epilepsy in adulthood. Study design We conducted a cohort study of 21,441 births with record linkage to data from the Tayside Medicine Monitoring unit (MEMO) and Scottish morbidity records (SMR1). RESULTS: Delivery by forceps was not associated with epilepsy compared with all other deliveries, adjusted odds ratio (OR) 1.0 (95 % CI, 0.6-1.8). Epilepsy in adulthood was associated with a family history of epilepsy, adjusted OR 2.4 (95% CI, 1.7-3.2), increasing social deprivation, adjusted OR 1.1 for each Carstairs score (95% CI, 1.0-1.2), and male gender, adjusted OR 1.4 (95% CI, 1.0-1.8). Preterm birth was associated with an increased risk of epilepsy, adjusted OR 2.0 (95% CI, 1.2-3.2) but no other antenatal, intrapartum, or neonatal risk factors were identified. CONCLUSION: These findings do not suggest an association between forceps delivery and epilepsy in adulthood; however, preterm birth may be an important risk factor. PMID- 15343213 TI - Prescription drug use in pregnancy. AB - OBJECTIVE: The purpose of this study was to provide information on the prevalence of the use of prescription drugs among pregnant women in the United States. STUDY DESIGN: A retrospective study was conducted with the use of the automated databases of 8 health maintenance organizations that are involved in the Health Maintenance Research Network Center for Education and Research on Therapeutics. Women who delivered of an infant in a hospital from January 1, 1996, through December 31, 2000, were identified. Prescription drug use according to therapeutic class and the United States Food and Drug Administration risk classification system was evaluated, with the assumption of a gestational duration of 270 days, with three 90-day trimesters of pregnancy, and with a 90 day period before pregnancy. Nonprescription drug use was not assessed. RESULTS: During the period 1996 through 2000, 152,531 deliveries were identified that met the criteria for study. For 98,182 deliveries (64%), a drug other than a vitamin or mineral supplement was prescribed in the 270 days before delivery: 3595 women (2.4%) received a drug from category A; 76,292 women (50.0%) received a drug from category B; 57,604 women (37.8%) received a drug from category C; 7333 women (4.8%) received a drug from category D, and 6976 women (4.6%) received a drug from category X of the United States Food and Drug Administration risk classification system. Overall, 5157 women (3.4%) received a category D drug, and 1653 women (1.1%) received a category X drug after the initial prenatal care visit. CONCLUSION: Our finding that almost one half of all pregnant women received prescription drugs from categories C, D, or X of the United States Food and Drug Administration risk classification system highlights the importance of the need to understand the effects of these medications on the developing fetus and on the pregnant woman. PMID- 15343214 TI - To the point: reviews in medical education teaching techniques. AB - Medical educators are faced with the difficult dilemma of decreased time available for teaching as a result of increased demands for clinical time. Yet few physicians have formal training in teaching methods. Educational research is reviewed to assist educators to select the best teaching technique to meet the learning objectives. PMID- 15343215 TI - Thrombophilia and pregnancy complications. AB - OBJECTIVE: This systematic review examines the strength of the association between thrombophilia and recurrent pregnancy loss and other serious obstetric complications. Study design Electronic databases and manual bibliography searches were used to identify studies evaluating the association between thrombophilia and pregnancy loss, preeclampsia, fetal growth retardation, and placental abruption. RESULTS: Thrombophilic disorders are associated with an increased risk of fetal loss in the majority of case control and cohort studies. The risk is increased throughout pregnancy, but may be higher in the second and third trimester. The common pathologic finding of placental infarction suggests unexplained fetal loss may result from uteroplacental insufficiency and thrombosis. Thrombophilic disorders are not consistently associated with preeclampsia, fetal growth retardation, or placental abruption. Preliminary data suggest prophylactic anticoagulation may improve outcome in thrombophilic women with unexplained recurrent fetal loss. CONCLUSION: Women with thrombophilia have an increased risk of pregnancy loss and possibly other serious obstetric complications, although definition of the magnitude of risk will require prospective longitudinal studies. Preliminary data suggesting prophylactic anticoagulation may improve gestational outcome provide a rationale for prospective randomized trials in thrombophilic women with unexplained recurrent fetal loss. PMID- 15343216 TI - Uterine rupture: differences between a scarred and an unscarred uterus. AB - OBJECTIVES: This study aimed to compare risk factors, site of rupture, and outcome of uterine rupture among patients with a scarred versus an unscarred uterus. Study design We conducted a comparison between all cases of uterine rupture (n=53) in women with a scarred versus an unscarred uterus, occurring between January 1988 and July 2002. RESULTS: During the study period, there were ruptures among 26 patients with a scarred uterus and 27 patients without a uterine scar. No significant differences were noted between the scarred and unscarred groups while comparing risk factors such as birth order, birth weight, hydramnios, oxytocin induction, diabetes, and malpresentation. The main site of involvement in both groups was the lower uterine segment representing 92.6% of the ruptures in the unscarred group and 92.3% of the ruptures in the scarred uterus group. Cervical involvement was significantly more common among patients without a previous uterine scar (33.3% vs 7.7%; odds ratio [OR]=6.0, 95% CI, 1.16 31.23, P=.04). Conversely, uterus corpus involvement did not differ between the groups. Perinatal mortality did not differ between the groups. In addition, no significant differences were noted regarding maternal morbidity such as the need for hysterectomy, blood transfusion, or length of hospitalization. CONCLUSION: Although cervical involvement was significantly more prevalent in the rupture of an unscarred uterus, no significant differences in maternal or perinatal morbidity were noted between rupture of a scarred versus an unscarred uterus. PMID- 15343217 TI - Colposcopically directed biopsy, random cervical biopsy, and endocervical curettage in the diagnosis of cervical intraepithelial neoplasia II or worse. AB - OBJECTIVES: The purpose of this study was to determine the relative importance of colposcopically directed biopsy, random biopsy, and endocervical curettage (ECC) in diagnosing > or =cervical intraepithelial neoplasia (CIN) II. Study design During a screening study, 364 women with satisfactory colposcopy and > or =CIN II were diagnosed. All colposcopically detected lesions were biopsied. If colposcopy showed no lesion in a cervical quadrant, a random biopsy was obtained at the squamocolumnar junction in that quadrant. ECC was then performed. RESULTS: The diagnosis of > or =CIN II was made on a colposcopically directed biopsy in 57.1%, random biopsy in 37.4%, and ECC in 5.5% of women. The yield of > or =CIN II for random biopsy when cytology was high grade (17.6%) exceeded that when cytology was low grade (2.8%). One of 20 women diagnosed solely by ECC had invasive cancer. CONCLUSION: Even when colposcopy is satisfactory, ECC should be performed. If cytology is high grade, random biopsies should be considered. PMID- 15343218 TI - Hysteroscopic injection of tracers in sentinel node detection of endometrial cancer: a feasibility study. AB - OBJECTIVE: The purpose of this study was to evaluate the feasibility of sentinel lymph node (SLN) detection in endometrial cancer patients with hysteroscopic injection of tracers. Study design Eighteen patients with endometrial adenocarcinoma were submitted to hysteroscopic injection of technetium-99m labeled colloids and blue-dye subendometrially around the lesion followed by lymphoscintigraphic scans. The SLN was detected by direct visualization of blue dye marked nodes and by a radio-guided surgery (RGS). RESULTS: Seventeen hysteroscopic procedures were satisfactory regarding the visualization of the uterine cavity. The compliance to the procedure was acceptable in 15 cases, with no severe complication. A hysterectomy and bilateral salpingo-oophorectomy were performed in all cases, and pelvic lymphadenectomy in 14 cases. The RGS detected a total of 45 SLN with a mean of 3 SLN/patient (range 2 to 4). Blue-dye uptake was observed in 6 (33%) cases. No case presented blue-dye uptake and radioactive colloid negativity. CONCLUSION: The results seem promising for further controlled studies. PMID- 15343219 TI - Assessing the variation in antiphospholipid antibody (APA) assays: comparison of results from 10 centers. AB - OBJECTIVE: This study was undertaken to determine the variability of antiphospholipid antibody (APA) assay results of cardiolipin and other frequently tested phospholipids. Study design Ten centers performing APA assays submitted samples that were positive in their assay. Twenty samples were identified to cover a broad range of APA results. Samples were distributed to the 10 participating centers for evaluation of immunoglobulin G (IgG), IgM, and IgA antibodies to phospholipids. RESULTS: Of 20 patients, 9 (45%) were identified as positive by all 10 centers and 2 of 20 patients (10%) were identified as positive by 8 or 9 centers. However, 9 of 20 samples (45%) returned with mixed results. The average percent of positive samples was 29%, but results from the 10 centers ranged from a low number of positives (13%) to a high number of (37%). CONCLUSION: When considering a clinical diagnosis of APA syndrome, laboratory concordance of results from 20 patient samples among the 10 centers was only 55%. However, when considering a single phospholipid of a single immunoglobulin isotype, agreement of test results was 83.8%. Isolated positive APA results should be correlated with the clinical history and confirmed by repeat testing. PMID- 15343220 TI - Use of a new platelet function analyzer to detect von Willebrand disease in women with menorrhagia. AB - OBJECTIVE: The purpose of this study was to assess the usefulness of a new platelet function analyzer in the detection of von Willebrand disease and platelet dysfunction in women with menorrhagia. STUDY DESIGN: Women with menorrhagia and control subjects were tested with a platelet function analyzer. If the results were abnormal, further testing was performed for possible von Willebrand disease or platelet dysfunction. Results were compared. RESULTS: Of the 108 women with menorrhagia, 28 had an abnormal platelet function analysis results: 7 results were suggestive of von Willebrand disease; 17 results were suggestive of platelet dysfunction, and 4 results were inconclusive. Of the 100 control subjects, 18 subjects had an abnormal platelet function analysis result: 2 results were suggestive of von Willebrand disease; 12 results were suggestive of platelet dysfunction, and 4 results were inconclusive. The prevalence of suspected von Willebrand disease was 6% among women with menorrhagia and 2% among control subjects. Among white women, the prevalence was 10% compared with 1% among control subjects. CONCLUSION: We demonstrated that the platelet function analyzer could be used to detect von Willebrand disease in women with menorrhagia. PMID- 15343221 TI - Racial and ethnic disparities in the provision of epidural analgesia to Georgia Medicaid beneficiaries during labor and delivery. AB - OBJECTIVE: The purpose of this study was to measure racial and ethnic differences in the proportion of Medicaid patients who receive epidural analgesia during labor and delivery. STUDY DESIGN: Using 1998 Georgia Medicaid claims data in a standard State Medicaid Research File format, we identified claims for epidural analgesia among all women who had a normal vaginal delivery during 1998. RESULTS: There were 29,833 women who met our inclusion criteria, of whom 15,936 (53.4%) had epidural analgesia. Epidural analgesia rates were lower for black women (49.5%), Hispanic women (35.3%), and Asian women (48.1%) than for white, non Hispanic women (59.6%; P<.001). Rural women had lower epidural rates (39.2%) than urban women (62.1%). CONCLUSION: The study subjects all had identical Medicaid insurance and met the same low-income Medicaid eligibility criteria, yet race/ethnicity was still a significant predictor of epidural analgesia after we had controlled for age, rural-urban residence, and availability of anesthesiologists. Further studies are needed to assess perceived benefits, risks, costs, and obstacles to epidural analgesia that are perceived by patients, physicians, nurses, and midwives. PMID- 15343222 TI - Parity and route of delivery: does cesarean delivery reduce bladder symptoms later in life? AB - OBJECTIVE: This study was undertaken to investigate the impact of reproductive factors on the prevalence of urinary symptoms. STUDY DESIGN: Participants were women scheduled for hysterectomy (n=1299). Before surgery, urinary symptoms were assessed by questionnaire. Multiple logistic regression analysis was used to investigate the association between bladder symptoms and parity, route of delivery, and other characteristics. RESULTS: Stress incontinence and urinary urgency were more prevalent among parous than nulliparous women (P <.01). Controlling for parity and other characteristics, women who had a history of cesarean delivery were significantly less likely to report stress incontinence than women with a history of vaginal delivery (odds ratio 0.60; 95% CI 0.39 0.93). CONCLUSION: Women who have undergone vaginal delivery are more likely to report stress incontinence than women who have delivered by cesarean section. Although this suggests that cesarean delivery might reduce incontinence later in life, further research is needed to clarify the long-term risks, benefits, and costs of cesarean delivery. PMID- 15343223 TI - Delayed interval delivery and infant survival: a population-based study. AB - OBJECTIVE: Delaying delivery of the remaining fetus(es) in a multifetal pregnancy is feasible in some cases. However, the impact of this procedure on infant survival is unclear. STUDY DESIGN: We used the US 1995-1998 Matched Multiple Birth File. We identified 200 twin pregnancies in which the first twin was delivered between 17 and 29 weeks of gestation and the second twin was delivered 2 or more days later. We individually matched the delayed deliveries with 374 twin pregnancies in which the second twin was delivered on the same or next calendar day. Perinatal outcomes and infant survival were compared between the delayed and nondelayed twins. RESULTS: Among the 200 pregnancies with delayed delivery, the mean gestational age at first delivery was 23 weeks and the median duration of delay was 6 days (ranging from 2-107 days). One week of delay in delivery was associated with an increase in infant birth weight of 131 g on average (95% CI: 115-147 g). Moreover, 56% of the delayed second twins survived to 1 year of age, whereas only 24% of the nondelayed second twins survived to 1 year of age (P <.001). However, 11% of the second twin in delayed delivery (95% CI: 6%-16%) experienced fetal death before 24 weeks. CONCLUSION: Delayed delivery of the remaining fetus(es) before 30 weeks of gestation for 2 or more days was associated with improved infant survival. PMID- 15343224 TI - Selective fetocide reverses preeclampsia in discordant twins. AB - OBJECTIVE: Preeclampsia occurs in some twin gestations in association with a placental disorder of one but not both fetuses, thereby placing both at risk. We investigated a novel method of treating preeclampsia in these pregnancies. STUDY DESIGN: Three patients with second trimester preeclampsia linked to a lethal condition in one twin were treated with selective fetocide in an effort to reverse preeclampsia. Two patients presented with twins discordant for severe fetal growth restriction, and 1 patient presented with Ballantyne syndrome and twins discordant for fetal hydrops. RESULTS: Preeclampsia resolved in all 3 patients, allowing continuation of the pregnancy for an additional 9 to 23 weeks before delivery of the remaining fetus. Resolution of preeclampsia occurred in a timeframe consistent with placental involution documented in similar clinical circumstances. CONCLUSION: Selective fetocide may be an option for treating preeclampsia in some twin pregnancies, presumably by causing involution of the pathologic placenta. Delivery is not the only cure for preeclampsia. PMID- 15343225 TI - Intrauterine growth restriction increases morbidity and mortality among premature neonates. AB - OBJECTIVE: Intrauterine growth restriction (IUGR) is an important reason for premature delivery and has been reported to be associated with increased mortality, but in some studies paradoxically, improved morbidities. Data on neonatal outcomes for infants with IUGR at each viable gestational age at birth from large numbers of deliveries are lacking. More particularly, data on perinatal outcome related to an antenatal diagnosis of IUGR compared with a neonatal diagnosis are particularly deficient. Therefore, by using a large contemporary database, we evaluated the outcomes of neonates with IUGR and the gestational age-specific associations between growth restriction, morbidity, and mortality. STUDY DESIGN: With the use of a database formed from a computer assisted tool that generates clinical progress notes and discharge summaries on neonatal intensive care unit (NICU) admissions, we reviewed data on neonates discharged from 124 NICUs between January 1, 1997, and December 31, 2001. We evaluated singleton, inborn neonates who delivered between 23 and 34 weeks, excluding major congenital anomalies. We compared 3 measures of IUGR: antenatally diagnosed IUGR; a birth weight below the 10th percentile (small for gestational age [SGA]), and newborn infants with either or both of these diagnoses against a control group of gestational age-matched infants meeting none of these criteria whose birth weights were no greater than the 90th percentile. RESULTS: Our sample included 29,916 prematurely born neonates; 1,451 (4.8%) with IUGR, 2,936 (9.8%) who were SGA, and 3,708 (12.3%) had at least 1 of these 2 markers. There were 22,798 (76%) normally grown control neonates. Within each gestational age group from 25 to 32 weeks, each marker of IUGR was associated with increased mortality, necrotizing enterocolitis, need for respiratory support at 28 days of age, and retinopathy of the premature. When corrected for gestational age, exposure to antenatal steroids, gender, and mode of delivery, these associations remained significant. CONCLUSION: IUGR remains a serious problem that is associated with increased morbidity and mortality among prematurely born neonates, regardless of the definition used or whether the diagnosis is made antenatally or after birth. These results are important for obstetric counseling and decision making and for the anticipation and treatment of premature newborn infants. PMID- 15343226 TI - Necrotic cell injury in the preterm and near-term ovine fetal brain after intermittent umbilical cord occlusion. AB - OBJECTIVE: This study was undertaken to determine the extent of necrotic cell death as a measure of neurologic injury in the preterm and near-term ovine fetal brain in response to intermittent umbilical cord occlusion (UCO) with severe, but limited hypoxia and no cumulative acidosis to ensure longer-term survival. STUDY DESIGN: Fetal sheep (control and experimental groups at 0.75 and 0.90 of gestation) were studied over 4 days with UCOs performed in the experimental group animals by complete inflation of an occluder cuff for 90 seconds every 30 minutes for 3 to 5 hours each day. Animals were then euthanized and the fetal brain perfusion-fixed and prepared for subsequent histology and assessment of necrotic cell injury by using standard staining with hematoxylin and eosin (H&E), and with a novel fluorescent marker, Fluoro-Jade B, that targets degenerating neurons. RESULTS: In both preterm and near-term animal groups, UCO caused a large decline in arterial Po(2) (to approximately 7 mm Hg), a modest decline in pH (to approximately 7.30), and a modest rise in Pco(2) (to approximately 61 mm Hg) (all P<.01), but with a return to control values after the occluder release and no cumulative acidosis over each day of study. Overall, very low levels of H&E stained necrotic-appearing cells and Fluoro-Jade B-stained positive cells were observed across all brain regions studied with values not significantly different from zero, excepting that for the gray matter of the preterm control (by H&E staining), preterm and near-term cord occlusion (by H&E and Fluoro-Jade B staining), and the thalamus of the near-term cord occlusion (by H&E staining) animals. Although there were no differences in the levels of H&E-stained necrotic appearing cells and Fluoro-Jade B-stained positive cells between respective control and cord occlusion group animals for most of the brain regions studied, a significant increase in Fluoro-Jade B-stained positive cells was observed in the gray matter of both the preterm and near-term cord occlusion animals (P<.05). CONCLUSION: Intermittent cord occlusion insult with severe but limited fetal hypoxemia and no cumulative acidosis, was generally well tolerated in both the preterm and near-term animal groups as assessed by measures of necrotic cell injury throughout the brain with minimal evidence for such. However, compensatory mechanisms which are protective for the brain may become limited with repetitive hypoxia insult over time as suggested by the low level of Fluoro-Jade B-stained positive cells noted in the gray matter tissues for both occlusion groups. PMID- 15343227 TI - Structural proteins during brain development in the preterm and near-term ovine fetus and the effect of intermittent umbilical cord occlusion. AB - OBJECTIVE: The purpose of this study was to determine the immunoreactivity of selected structural proteins in the preterm and near-term ovine fetal brain and the response to intermittent umbilical cord occlusion as a measure of altered cellular growth. The intermediate filament proteins nestin, vimentin, and glial fibrillary acidic protein was used as markers for astroglial maturation and astrogliosis, and myelin basic protein as a marker for oligodendrocytes and myelin formation. STUDY DESIGN: Fetal sheep (control and experimental groups at 0.75 and 0.90 of gestation) were studied over 4 days; umbilical cord occlusion was performed in the experimental group by complete inflation of an occluder cuff for 90 seconds every 30 minutes for 3 to 5 hours each day. Animals were then killed, and the fetal brain was perfusion fixed and processed for immunohistologic examination of the gray and white matter. Immunoreactivity was quantified with an image analysis system and expressed as the fractional area positive stain for each protein. RESULTS: In both preterm and near-term animal groups, umbilical cord occlusion caused a large decline in arterial Po(2) (to approximately 7 mm Hg), a modest decline in pH (to approximately 7.30), and a modest rise in Pco(2) (to approximately 61 mm Hg; all P <.01), with a return to control values after the occluder release and no cumulative acidosis over each day of study. Vimentin and glial fibrillary acidic protein immunoreactivity showed reciprocal changes, with vimentin decreased and glial fibrillary acidic protein increased in both the gray and white matter of the control group from 0.75 to 0.90 of gestation, which can be attributed to the transition of radial glia into mature astrocytes. Myelin basic protein immunoreactivity increased approximately 3-fold in the white matter of the control group with advancing gestation, which likely reflected active oligodendrocyte differentiation and increased myelination at this time of development. Intermittent umbilical cord occlusion over 4 days resulted in an approximately 60% decrease in nestin, vimentin, and glial fibrillary acidic protein immunoreactivity, which was qualitatively similar for both the gray and white matter and likely indicative of altered protein synthesis and/or degradation, but only in the preterm group and with no change in myelin basic protein immunoreactivity. CONCLUSION: There is considerable change in the immunoreactivity of structural proteins within the ovine fetal brain over the latter part of gestation and consistent with a high rate of protein turnover, as previously reported. Intermittent umbilical cord occlusion as studied with minimal evidence for necrotic cell injury appears capable of altering selected protein synthesis/degradation, more so in younger animals when protein turnover is higher, which might then impact on the brain's development. PMID- 15343228 TI - The effect of advancing paternal age on pregnancy and live birth rates in couples undergoing in vitro fertilization or gamete intrafallopian transfer. AB - OBJECTIVE: This study was undertaken to determine effects of male aging on sperm parameters, fertilization, pregnancy, and live birth rates among in vitro fertilization (IVF) or gamete intrafallopian transfer (GIFT) couples. The impact of female age was also investigated. STUDY DESIGN: Prospective study was made up of 221 IVF and GIFT couples. RESULTS: Pregnancy rates declined as the male subjects aged. Each additional year of paternal age was associated with 11% increased odds (P=.007) of not achieving a pregnancy, and 12% odds (P=.01) of not having a successful live birth. For first-time IVF/GIFT recipients, each additional year of paternal age was associated with a 5% increased odds of not achieving a pregnancy, whereas for repeaters it was 40% (P=.01). Advancing maternal age was associated with decreased numbers of oocytes retrieved or fertilized. Women 40 years or older compared with younger than 35 years had greater than 4-fold risk of not becoming pregnant, and greater than 20-fold risk of not achieving a live birth. CONCLUSION: Advancing paternal (and maternal) age had a deleterious effect on IVF and GIFT outcomes. PMID- 15343229 TI - Increased fetal DNA in the maternal circulation in early pregnancy is associated with an increased risk of preeclampsia. AB - OBJECTIVE: The aim of our study was to determine if fetal DNA is present in the maternal circulation in early pregnancy before the clinical manifestation of preeclampsia, and if this could be predictive of the development of preeclampsia. STUDY DESIGN: Blood were obtained from patients attending for a first antenatal visit. Cases were asymptomatic women who subsequently developed preeclampsia matched to control women for parity and gestational age. Real-time polymerase chain reaction (PCR) using TaqMan primers and probes directed against SRY gene sequences quantified fetal DNA in the maternal circulation. RESULTS: There were 88 cases of women with preeclampsia and 176 control women, both sampled at a mean gestation (+/-SD) of 15.7 +/- 3.6 weeks. The presence of fetal DNA in the maternal circulation in early pregnancy is associated with an 8-fold increased risk of developing preeclampsia. CONCLUSION: Increased fetal DNA is present in the maternal circulation in early pregnancy in women who subsequently develop pre eclampsia and there appears to be a graded response between the quantity of fetal DNA and the risk of developing pre-eclampsia. PMID- 15343230 TI - Vaginal versus ultrasound examination of fetal occiput position during the second stage of labor. AB - OBJECTIVE: The purpose of this study was to determine whether ultrasonography is more accurate than vaginal examination in the determination of fetal occiput position in the second stage of labor. STUDY DESIGN: Eighty-eight patients in the second stage of labor were evaluated by vaginal examination and by combined transabdominal and transperineal ultrasound examination to determine occiput position. These predictions of position were compared with the actual delivery position at vaginal delivery after spontaneous restitution or at cesarean delivery. Different examiners performed the vaginal examinations and the ultrasound examinations. Each examiner was blinded to the determination of the other examiner. RESULTS: Vaginal examination determined fetal occiput position correctly 71.6% of the time; ultrasound examination determined fetal occiput position correctly 92.0% of the time (P=.018). CONCLUSION: Ultrasound examination is more accurate than vaginal examination in the diagnosis of fetal occiput position in the second stage of labor. PMID- 15343231 TI - HLA-G protein concentrations in maternal serum and placental tissue are decreased in preeclampsia. AB - OBJECTIVE: The aim of this study were to determine whether soluble human leukocyte antigen-G protein levels in serum and/or human leukocyte antigen protein in placental tissues differ between women with preeclampsia versus uncomplicated pregnancies. Study design human leukocyte antigen-G levels were determined with the use of a specific enzyme-linked immunosorbent assay in 20 subjects with preeclampsia and 14 normal control subjects. RESULTS: Both serum and placental human leukocyte antigen-G levels were decreased significantly in the preeclampsia group (median, 0.026 microg/mL in serum; median, 0.026 microg/mg protein in placenta), in comparison with normal pregnant women (median, 0.093 microg/mL in serum; median, 0.088 microg/mg protein in placenta; P=.0112 and P=.0406, respectively). There was a significant correlation between serum and placental human leukocyte antigen-G levels (r=0.603; P=0.0002). CONCLUSION: The reduced expression of placental human leukocyte antigen-G and reduced release of this protein into the maternal circulation in preeclampsia may alter the maternal fetal immune relationship and thus be involved in the cause of this disorder. PMID- 15343232 TI - Efficacy of an intervention to prevent excessive gestational weight gain. AB - OBJECTIVE: This study was undertaken to evaluate the efficacy of an intervention directed at preventing excessive gestational weight gain. STUDY DESIGN: Healthy pregnant women with normal and overweight early pregnancy body mass index were monitored from early pregnancy until 1-year postpartum. One hundred seventy-nine women in the intervention group had their gestational weight gains monitored by health care providers and also received by-mail patient education. Three hundred eighty-one women formed an historical control group. The proportions in each group gaining more weight in pregnancy than is recommended and retaining more than 2.27 kg at 1-year postpartum were compared using logistic regression analysis. RESULTS: Low-income women who received the intervention had a significantly reduced risk of excessive gestational weight gain (odds ratio [OR]=0.41, 95% CI=0.20-0.81). Overweight women within this income subgroup were at significantly reduced risk of retaining more than 2.27 kg (OR=0.24, 95% CI=0.07-0.89). CONCLUSION: The intervention appeared to reduce the risk of excessive gestational weight gain only in the low-income subgroup. PMID- 15343233 TI - Serum homocysteine levels after preterm premature rupture of the membranes. AB - OBJECTIVE: Preterm premature rupture of the membranes (PPROM) is believed to be caused, in part, by abnormalities of collagen and increased levels of oxidative stress. Elevated homocysteine levels have been shown to induce these same pathophysiologic changes. We tested the hypothesis that serum homocysteine levels would be higher in women with PPROM when compared with matched control women. STUDY DESIGN: A secondary analysis derived from 2 previously completed studies performed in the National Institutes of Child Health and Human Development Maternal-Fetal Medicine Units (MFMU) Network. We identified 99 study cases with PPROM (24 to 32 weeks' gestation) and matched them with 99 asymptomatic control women from an observational study of preterm birth prediction. Cases and control women were matched for race, gestational age at sampling, and MFMU Network center. Serum homocysteine levels were determined by immunoassay in batch fashion by personnel masked to study arm and clinical outcomes. Serum homocysteine levels were compared between groups, as were the baseline characteristics of maternal age, cigarette smoking, nulliparity, infections during pregnancy, and body mass index (BMI) <19.8 kg/m 2. Serum homocysteine levels were dichotomized as >75th, 90th, and 95th %ile of control women, and the likelihood of elevated homocysteine levels was determined in women who smoked, had a BMI <19.8 kg/m 2, or who had PPROM. Statistical analyses included the Wilcoxon rank sum, chi-square, and Pearson correlation coefficient, where appropriate. Baseline characteristics were controlled with a logistic regression model. RESULTS: Serum homocysteine levels measured in patients with PPROM were not significantly different from matched control women: median and (25th to 75th %ile): 4.9 (3.5-6.2) vs 4.8 (3.9-6.2 micromol/L), P =.73. In our population, neither the number of cigarettes smoked ( r = -0.08, P =.57), nor BMI ( r = -0.08, P =.24) correlated with serum homocysteine levels. The strongest association was seen in women with PPROM having serum homocysteine levels >95th %ile of control women (odds ratio [OR] 2.7, P =.10). After adjusting for baseline characteristics, no correlation between serum homocysteine level and the presence of PPROM was seen, OR 1.0 (.9 1.1); P =.99. CONCLUSION: Women presenting with PPROM did not have significantly increased serum homocysteine levels when compared with control women. PMID- 15343234 TI - Clinical correlates of pain with amniocentesis. AB - OBJECTIVE: The purpose of this study was to determine whether sensory or affective dimensions of pain with genetic amniocentesis are associated with identifiable clinical correlates. STUDY DESIGN: Women completed the short-form McGill Pain Questionnaire after second-trimester genetic amniocentesis. The effect of maternal weight, parity, previous amniocentesis, previous surgery, history of menstrual cramps, maternal anxiety, presence of fibroid tumors, and depth and location of needle insertion on pain intensity was determined. The T test, correlation matrix, Kruskal-Wallis test, and multiple logistic regression were used for analysis; a probability value of <.05 was considered significant. RESULTS: One hundred twenty-one women were enrolled: 19.3% reported no pain, 42.9% described the pain as mild, 31.1% described the pain as discomforting, and 6.7% described the pain as distressing or horrible. Mean intensity of pain was 1.6+/-1.3 (on a scale of 0-7). Pain was most often described as sharp, cramping, fearful, and stabbing. Anxiety and pain were increased in women with an indication of abnormal serum screen as compared with women with advanced maternal age. Anxiety and a history of menstrual cramps were associated with increased affective dimensions of pain and had moderate correlation with quantified pain intensity. A history of previous amniocentesis and needle insertion in the lower one third of the uterus were associated with increased pain. Maternal weight, parity, previous surgery, fibroid tumors, and depth of needle insertion were not correlated with perceived pain. Presence or absence of an accompanying person was not associated with pain intensity. CONCLUSION: Women report mild pain or discomfort with genetic amniocentesis. Increased pain is associated with increased maternal anxiety, a history of menstrual cramps, a previous amniocentesis, and insertion of the needle in the lower uterus. PMID- 15343235 TI - Screening for post-miscarriage psychiatric morbidity. AB - OBJECTIVE: The purpose of this study was to evaluate 12-item General Health Questionnaire (GHQ-12) in screening for psychiatric morbidity after miscarriage. STUDY DESIGN: A prospective cohort study was carried out involving 222 patients. Six weeks after miscarriage, the GHQ-12 was applied. Psychiatric "case" or "non case" was diagnosed by the psychiatrist with use of the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders-III-R. The patients were computer randomized into Groups A or B. A receiver operating characteristic (ROC) curve was constructed for Group A. The optimal cutoff value of GHQ-12 was determined, and this value was applied to Group B. The test characteristics were assessed. RESULTS: Twenty-seven patients were found to be psychiatric cases. An ROC with area under curve of 0.93 (95% CI 0.87-0.99, P<.001) was constructed. The best GHQ-12 cutoff score was > or =4 in detecting psychiatric caseness. A sensitivity of 83%, specificity of 90%, positive predictive value of 50%, and negative predictive value of 98% were obtained. CONCLUSION: GHQ-12 is an effective screening tool in detecting psychiatric morbidity after miscarriage. PMID- 15343236 TI - Quantification of fetomaternal hemorrhage: a comparative study of the manual and automated microscopic Kleihauer-Betke tests and flow cytometry in clinical samples. AB - OBJECTIVE: The purpose of this study was to evaluate the quantification of fetomaternal hemorrhage by the manual and automated microscopic analysis of Kleihauer-Betke stained slides and by flow cytometry. STUDY DESIGN: Blood smears were stained and evaluated manually according to the Kleihauer-Betke test. The same slides were used for automated microscopy. In addition, blood flow cytometry was performed by anti-hemaglobin F immunostaining. RESULTS: Fetomaternal hemorrhage >0.1% was detected in 4 patients by manual and automated Kleihauer Betke test and by blood flow cytometry. Fetomaternal hemorrhage was absent according to all 3 methods in 13 patients; fetomaternal hemorrhage<0.1% was detected in 27 patients by either manual or automated Kleihauer-Betke test or both. Moderate agreement was observed between the manual and automated Kleihauer Betke test (weighted kappa, 0.56; 95% CI, 0.33-0.78). Agreement between the manual Kleihauer-Betke test and blood flow cytometry was fair (weighted kappa, 0.40; 95% CI, 0.15-0.66). CONCLUSION: Automated microscopic detection of fetal blood cells in clinical samples provides accurate quantification that is comparable to the manual Kleihauer-Betke test in both small and large fetomaternal hemorrhage. Blood flow cytometry is capable only of quantifying fetomaternal hemorrhage of >0.1%. PMID- 15343237 TI - Maternal-fetal transfer and amniotic fluid accumulation of protease inhibitors in pregnant women who are infected with human immunodeficiency virus. AB - OBJECTIVE: The purpose of this study was to investigate placental transfer and amniotic fluid concentrations of protease inhibitors when they are given to pregnant women who are infected with human immunodeficiency virus. STUDY DESIGN: Fifty-eight mothers who received antiretroviral therapy that included > or =1 protease inhibitors for clinical indications at the time of delivery were enrolled in the study. Maternal blood samples and amniotic fluid were obtained during delivery or cesarean delivery, and paired cord blood samples were obtained by venipuncture immediately after the delivery. Drug concentrations were measured with high performance liquid chromatography. RESULTS: Most maternal protease inhibitor plasma concentrations (38/66 concentrations) were below the trough concentrations that are recommended for therapeutic drug monitoring. Cord blood concentrations were below the assay limit of detection in 10 of 40 samples for nelfinavir and 25 of 40 samples for its metabolite M8, 9 of 11 samples for ritonavir, 4 of 6 samples for indinavir, 5 of 6 samples for saquinavir but were detectable in 3 of 3 samples for amprenavir. Among the 24 amniotic fluid samples that were available, the concentrations below the detection limit were 10 of 16 samples for nelfinavir, 11 of 16 samples for M8, 1 of 3 samples for indinavir, 4 of 4 samples for ritonavir, and 0 of 1 samples for amprenavir. There were significant correlations between cord blood and maternal concentrations of nelfinavir and its metabolite M8. CONCLUSION: Placental transfer of the human immunodeficiency virus protease inhibitors is generally low; however, it may differ greatly according to the molecule. PMID- 15343238 TI - Homocysteine thiolactone induces apoptosis in cultured human trophoblasts: a mechanism for homocysteine-mediated placental dysfunction? AB - OBJECTIVE: Hyperhomocystinemia is a thrombophilic condition associated with placental dysfunction. We tested the hypothesis that homocysteine-thiolactone, a metabolite of homocysteine, induces apoptosis in cultured trophoblasts. STUDY DESIGN: Cytotrophoblasts from term human placentas were cultured for 72 hours or less in the presence or absence of 50 to 400 micromol/L homocysteine-thiolactone or 400 micromol/L cysteine (control), with or without vitamin C, vitamin E, folate, or N-acetylcysteine. Cell death was assessed by cellular adenosine triphosphate concentration, medium lactate dehydrogenase level, and immunocytochemical staining for the cleavage products of cytokeratin 18 and poly(adenosine diphosphate ribose) polymerase. Changes in expression of p53, Bcl 2, Bax, and Bak were quantified by Western immunoblotting. RESULTS: Homocysteine thiolactone induced a concentration dependent increase in total cell death and death by apoptosis, compared with control. Vitamin C ameliorated apoptosis in cytotrophoblasts, whereas N-acetylcysteine mitigated cell death in syncytiotrophoblasts. Apoptosis in both phenotypes occurred with increased expression of p53 and Bak, but no change in Bcl-2 or Bax. CONCLUSION: Homocysteine-thiolactone enhances apoptosis in cultured human trophoblast, and the effect can be limited by antioxidants. PMID- 15343239 TI - L-arginine erythrocyte transport increases during pregnancy and immediately postpartum. AB - OBJECTIVE: This study was undertaken to evaluate erythrocyte membrane transport of L-arginine in pregnancy and immediately postpartum. STUDY DESIGN: The study comprised 103 women with normal pregnancy, initially evaluated at the second trimester (II), followed into the third trimester (III), and immediately postpartum (PP). Total erythrocyte L-arginine uptake was measured with (14)C-L arginine, at 37 degrees C, for 3 minutes. The maximal transport capacity (V(max)) and half-saturation constant (K(m)) were obtained with the use of Michaelis Menten kinetics. Results are expressed as mean+/-SD. Analysis of variance, followed by Tukey test, was used in statistical analysis (alpha< or =.05). RESULTS: V(max) (micromol/L cells per hour) progressively increased at each consecutive time period: 779+/-283, 946+/-289, and 1349+/-390, at II, III, and PP, respectively (P<.001). Similarly, K(m) (micromol/L) values increased from 56+/-20 at time II, to 62+/-18 at time III, and 69+/-24 at PP (P<.001). CONCLUSION: Total erythrocyte L-arginine uptake (V(max) and K(m)) increases progressively along normal pregnancy, with a further increase immediately postpartum. PMID- 15343240 TI - The postprandial glucose profile in the diabetic pregnancy. AB - OBJECTIVE: A controversy exists regarding the time to monitor blood glucose in the diabetic pregnancy (60 or 120 minutes after meals). Using a novel approach that provides continuous measurement of blood glucose, we sought to determine postprandial glucose profile in the diabetic pregnancy. STUDY DESIGN: Subjects were connected to a continuous glucose monitoring system for 72 consecutive hours. A continuous glucose monitoring system measures the interstitial glucose levels in subcutaneous tissue every 5 minutes. Women were instructed to record the time of each meal during the study period. For each meal, the first 240 minutes were analyzed. RESULTS: Sixty-five women participated in the study: 26 women were treated by diet alone; 19 women received insulin therapy, and 20 women had type 1 diabetes mellitus. The time interval from meal to peak postprandial glucose levels was similar in all the evaluated types of diabetic pregnancies and in good and poor control insulin-treated patients with gestational diabetes mellitus (approximately 90 minutes). Failure to return to preprandial glucose values within a 3-hour observation period was identified in approximately 50% of the patients. A similar postprandial glucose peak time was obtained for breakfast, lunch, and dinner in all study groups. Postprandial hypoglycemia events were noted in approximately 10% of the meals and occurred about 160 minutes after mealtime. CONCLUSION: The time interval for postprandial glucose peak in diabetic pregnancies is approximately 90 minutes after meals throughout the day and is not affected by the level of glycemic control. This information should be considered in the treatment of diabetes mellitus in pregnancy. PMID- 15343241 TI - Assessment of resident competency in the performance of sonohysterography: does the level of training impact the accuracy? AB - OBJECTIVE: The objective of this study was to examine the true-positive rates by level of training of the operator and to determine whether the accuracy of the procedure differed by the level of education after formalized training. STUDY DESIGN: This was a retrospective analysis. The records of patients who underwent surgery for the indication of an abnormal saline sonohysterography were reviewed from January 2001 to April 2003 (n=73 patients). The nature of the saline sonohysterography abnormality, the intraoperative findings, and the level of training of the provider were recorded. Findings at saline sonohysterography were compared with findings at hysteroscopy or surgery. Statistical significance was determined by chi(2) test. RESULTS: The overall true-positive rate was 86.3% (63/73 patients). The true-positive rates for nurse practitioners, second- and fourth-year residents, and fellows were 84%, 80%, 90%, and 89%, respectively. There was no significant difference among providers (P=.96). CONCLUSION: The true positive rates for saline hysterography were comparable among different provider levels. PMID- 15343242 TI - The slippery slopes of advanced reproductive technologies: Presidential address. PMID- 15343244 TI - Qualitative human chorionicgonadotropin testing of cervicovaginal washings for the detection of preterm premature rupture of membranes. AB - OBJECTIVE: This study was performed to determine whether a qualitative human chorionicgonadotropin (hCG) test using cervicovaginal washings obtained from pregnant women is a useful predictor of recent preterm premature rupture of the membranes (PPROM). STUDY DESIGN: A prospective observational analysis of qualitative hCG testing on cervicovaginal washings in patients with either known PPROM or with intact membranes was performed. Patients with singleton gestations between 24 and 34 completed weeks were included. PPROM patients had documented confirmatory tests, including visualized pooling of amniotic fluid in the vaginal vault that was nitrazine-positive and demonstrated ferning. Those with vaginal bleeding or an anomalous fetus were excluded. Before collection, all patients were screened for bacterial vaginosis and trichomoniasis. Washings were then collected from the posterior vaginal fornix with the use of a 5-mL sterile saline irrigation and aspiration technique. Samples were then agitated manually and applied to the Quickvue Onestep qualitative pregnancy test (Quidell, San Diego, Calif) with a threshold of 25 mIU/mL. Data were analyzed with the t test, chi 2, and Fischer exact tests. Sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values were calculated. When applicable, all 2-tailed P <.05 were considered significant. RESULTS: Fifty-two patients were enrolled in the study with 24 in the PPROM group and 28 in the control group. The 2 groups were similar with respect to demographic characteristics and wet preparation results. The mean gestation age was 31.4 +/- 2.4 (X +/- SD) and 30.8 +/- 2.7 weeks in the PPROM and control groups, respectively. The hCG test was positive in 19 (79%) of the PPROM patients and in 1 (3.6%) of the controls (sensitivity 79%, specificity 96%, PPV 95%, NPV 84%). CONCLUSION: Qualitative hCG testing of cervicovaginal washings appears to be an useful predictor of PPROM. PMID- 15343246 TI - Results of in vitro fertilization in patients with endometriomas: is surgical removal beneficial? AB - OBJECTIVE: The purpose of this study was to investigate the effect of endometriomas on clinical pregnancy rates of in vitro fertilization-embryo transfer. STUDY DESIGN: Infertile patients with endometriosis who underwent in vitro fertilization-embryo transfer were divided into group 1 patients who had a history of ovarian endometriomas and group 2 patients who did not. The patients in group 1 were further divided into group 1A who had their endometriomas removed by laparoscopic ovarian cystectomy before the start of in vitro fertilization embryo transfer and group 1B who underwent in vitro fertilization-embryo transfer with the presence of endometriomas. The clinical pregnancy rates of these groups were compared. RESULTS: Clinical pregnancy rates per transfer were similar in group 1 and group 2 (40% vs 47%; P =.38) and similar in patients in group 1A and group 1B (47% vs 34%; P =.28), although the direction was toward improved pregnancy rates in the groups of patients without endometriomas present at the time of in vitro fertilization stimulation. When patients >39 years of age were excluded, the overall pregnancy rate was higher in those patients who had no current endometriomas than in those patients who had endometriomas present at the time of stimulation (65% vs 39%; P =.05). For patients with unilateral endometriomas, similar numbers of mature oocytes were aspirated between the affected and unaffected ovaries. CONCLUSION: Although the history of endometriomas does not appear to affect in vitro fertilization-embryo transfer outcomes adversely, there may be a benefit in the removal of endometriomas before in vitro fertilization. PMID- 15343248 TI - Diagnostic mid trimester amniocentesis: how safe? AB - OBJECTIVE: Genetic mid-trimester amniocentesis is a common invasive procedure. The origin of the commonly quoted 0.5% rate of procedure-related pregnancy loss after amniocentesis is obscure and is in conflict with the only randomized prospective study that reported a procedure-related rate of loss of 1.0%. This review was performed to establish an estimate of procedure-related pregnancy loss after mid-trimester amniocentesis. The impact of placental puncture on the rate of loss and the risk of direct needle injury to the fetus were also examined. STUDY DESIGN: The National Library of Medicine database was used to identify English language reports of >1000 amniocenteses with sufficient detail and follow up data to allow the calculation of the rate of spontaneous pregnancy loss after amniocentesis but before 28 completed weeks. These reports were divided into 2 groups to assess the impact of the ultrasound technique. Group I described only preamniocentesis ultrasound evaluation; group II described primarily concurrent ultrasound needle guidance. Pregnancy loss between the 2 groups was compared. The impact of placental puncture and reported direct fetal trauma were examined. The significance of differences was tested using chi-square analysis, with significance at a probability value of < or =.05. RESULTS: Twenty-nine reports that totaled 68,119 amniocenteses were examined. In a comparison of all studies in group I with all studies in group II, there was a lower rate of loss after amniocenteses with the use of concurrent guidance (1.4%) compared with the use of preamniocentesis ultrasound evaluation (2.1%) that was significant ( P <.001). Among only the 5 controlled studies that used preamniocentesis ultrasound evaluation, the difference in rate of loss between amniocentesis patients and control subjects was 0.6% ( P =.0042; 95% CI, 0.19, 1.03), which was identical to the difference in the rate of loss of 0.6% between amniocentesis patients and control subjects from the 5 controlled studies that used concurrent ultrasound needle guidance ( P <.0001; 95% CI, 0.31, 0.90). Multiple case reports and small series of presumed fetal needle trauma were reviewed, but most of these attributed causation to the amniocentesis needle based only on circumstantial association. Two cases with direct evidence of fetal needle trauma are discussed. Finally, the rate of loss after placental puncture from among 9 reports that provided this detail was 1.4% and not different from the overall rate of loss that was noted in group II. CONCLUSION: This examination of experience with 68,119 amniocenteses from both controlled and uncontrolled studies provides a substantive basis for several conclusions: (1) Contemporary amniocentesis with concurrent ultrasound guidance in controlled studies appears to be associated with a procedure-related rate of excess pregnancy loss of 0.6% (95% CI, 0.31, 0.90). To determine the total rate of loss, this must be added to the reported natural rate of loss without amniocentesis among control patients of 1.08%. (2) The use of concurrent ultrasound guidance appears to reduce the number of punctures and the incidence of bloody fluid. Concurrent ultrasound guidance was associated with a reduced rate of loss when all studies were compared, but not among controlled studies. (3) Direct fetal needle trauma is rare, and rarely proved, but may occur more frequently than is reported because of a failure to diagnose and a failure of the consistent production of sequelae. (4) This experience does not substantiate an increased rate of pregnancy loss if placental puncture is required. PMID- 15343249 TI - On prediction of the cesarean delivery risk in a large private practice. AB - OBJECTIVE: This study was undertaken to develop an easily usable integrated formula for predicting the probability of cephalopelvic disproportion/failure to progress (CPD) and cesarean delivery (CS) as a function of demographic factors in a middle-class private practice, and to evaluate risk factors for CS in a low risk primiparous population. METHODS: We studied 3355 primiparous women who delivered singleton births between February 1993 and July 2001 in a large private practice. We calculated body mass index (BMI) and weight gain during pregnancy by using clinical data from a comprehensive clinical database. Multivariable logistic regression analysis was used to estimate the relationship between the probability of CS and CS carried out for CPD and 6 demographic factors: maternal age, maternal height, initial pregnancy BMI, pregnancy weight gain, gestational age, and birth weight. Three methods were used to assess the accuracy of the model: the Hosmer-Lemeshow goodness-of-fit statistic, association of predicted probabilities, and direct comparison of the formula to the actual data. Odds ratios with a 95% CI are also calculated for each of these factors. RESULTS: The overall rate of primiparous CS for the practice is 21.7%, with 11.7% carried out for CPD. Formulas were developed to predict the probability of CS and the probability of CS caused by CPD. Our analysis shows that both the risk of CS and the risk of CS caused by CPD are significantly associated with all 6 demographic factors. We also develop an easily usable Web page-based calculator to instantly estimate any woman's probability of a CS or CPD at the beginning or at the end of her pregnancy. CONCLUSION: The probability of CS and CS performed for CPD is higher for shorter, older, more obese women with large pregnancy weight gains, larger fetal birth weights, and longer gestation ages. Accurate formulas that predict the probabilities of CS and CS performed for CPD in this large private practice have been developed. PMID- 15343251 TI - Estrogen receptor-alpha messenger RNA variants that lack exon 5 or exon 7 are coexpressed with wild-type form in human endometrium during all phases of the menstrual cycle. AB - OBJECTIVE: We have assessed the expression levels of messenger RNA for estrogen receptor-alpha and splice variants lacking exon 5 or exon 7 that presumably exert dominant positive (splice variants lacking exon 5) and negative (splice variants lacking exon 7) effects, respectively, on estrogen responses in the human endometrium. STUDY DESIGN: This was a prospective study that was conducted at an academic community-based hospital. The patients, aged 18 to 40 years, underwent hysterectomy for benign gynecologic causes. Eighty-one endometrial specimens (46 proliferative, 35 secretory) were analyzed with the use of reverse transcription polymerase chain reaction assay for the messenger RNA levels of estrogen receptor alpha, and splice variants lacking exon 5 and exon 7. RESULTS: Wild-type estrogen receptor-alpha and splice variants splice variants lacking exon 5 and lacking exon 7 messenger RNAs were detected in all endometrial specimens throughout the menstrual cycle. In addition, a double-splice estrogen receptor-alpha messenger RNA variant (splice variants lacking exon 5 and exon 7) was detected at constant low levels of expression. Semiquantitative analysis showed higher levels of estrogen receptor-alpha messenger RNA in the early and mid proliferative endometrial phases than in late proliferative and secretory endometrium ( P <.05). The splice variant lacking exon 7 messenger RNA expression level was about 10-fold higher than the splice variant lacking exon 5 messenger RNA relative to wild-type estrogen receptor-alpha messenger RNA ( P <.001). The expression of splice variants lacking exon 5 compared with wild-type estrogen receptor-alpha messenger RNA is relatively constant throughout endometrial development. In contrast, an examination of the ratio of the levels of splice variants lacking exon 7 to wild-type estrogen receptor-alpha messenger RNA indicated a small, but significantly higher, splice variant lacking exon 7 level in the mid secretory phase (postovulatory days 5-8) than the mid proliferative and early secretory phases ( P <.05). CONCLUSION: We found no evidence of differential coexpression of the positive dominant estrogen receptor variant, splice variants lacking exon 5, with wild-type estrogen receptor-alpha. We did find that the dominant negative splice variant lacking exon 7 was slightly increased relative to wild-type estrogen receptor-alpha in the postovulatory phase. Future investigation is required to suggest the biologic significance of the observed increased relative expression of the splice variants lacking exon 7 in secretory endometrium and to determine the function of splice variants lacking exon 5 and splice variants lacking exon 7. PMID- 15343253 TI - Clinical opinion: guidelines for hysterectomy. AB - OBJECTIVE: Abdominal hysterectomy remains the predominant method of uterine removal in the United States, despite evidence that vaginal hysterectomy offers advantages in regard to operative time, complication rates, return to normal activities, and overall cost of treatment. STUDY DESIGN: The predominance of the abdominal approach may be based on factors other than clinical considerations that include resident training, use of obsolete or limited guidelines, a perception rather than a confirmation that pathologic conditions exist that may suggest contraindications to the vaginal approach, misconceptions regarding the cost and safety of vaginal hysterectomy, and increased third-party reimbursement for the abdominal procedure. RESULTS: Evidence-based practice guidelines that were developed by the Society of Pelvic Reconstructive Surgeons and were adopted by the National Guidelines Clearinghouse have demonstrated that, in a number of studies that span several years, a dramatic shift toward the vaginal approach occurred when the guidelines were applied prospectively. CONCLUSION: The guidelines demonstrate that transvaginal hysterectomy is both feasible and optimal for many patients who long have been considered inappropriate candidates for vaginal hysterectomy. This clinical opinion attempts to address the reasons for the predominant use of the abdominal approach. PMID- 15343254 TI - Current opinions and understandings of menopausal women about hormone replacement therapy (HRT)-the University of Florida experience. AB - OBJECTIVE: This study was designed to assess the opinions and understanding of patients about hormone replacement therapy (HRT) since the release of the Women's Health Initiative (WHI) HRT trial data, which was widely publicized in the lay press. METHODS: All patients between the ages of 45 and 65 years seen in the last 3 years in the University of Florida Women's Clinics were mailed a survey asking about their attitudes and perceptions of HRT. RESULTS: A total of 6468 surveys were mailed with the return of 1076 completed surveys (16.6%); 78% of the respondents were menopausal and 65% had taken HRT. Of the women taking HRT, 70% stated it was for symptom relief and 30% for long-term benefits, including protection from heart disease (10%), Alzheimer's (4%), and osteoporosis (16%). Attitude changes about HRT since the release of WHI results were reported in 42% of respondents. Women currently on HRT had a more positive view of therapy (88%) compared with respondents overall (58%). Despite a great deal of media attention given to the topic of HRT, many women dramatically overestimated the risks of HRT, with 31% believing HRT increases the risk of heart disease 10% to 30% per year and 53% believing HRT increases the risk of breast cancer 10% to 30% per year. Despite this overestimation of risks, only 35% of respondents would not recommend HRT to a friend. CONCLUSION: This study suggests that WHI results did not impact attitudes about HRT in the majority of women taking HRT, as they were usually on therapy for relief of menopausal symptoms. However, there is a great need for consumer education about the attributable risks of HRT, approximately 1 per 1000 per year for heart disease, stroke, DVT, and breast cancer, so they can better weigh the benefits of HRT versus the risks. PMID- 15343256 TI - A randomized, multicenter study comparing the efficacy of recombinant FSH vs recombinant FSH with Ganirelix during superovulation/IUI therapy. AB - OBJECTIVE: The purpose of this study was to determine if use of a Gonadotropin releasing hormone (GnRH) antagonist, Ganirelix (Antagon), can improve pregnancy rates during superovulation with recombinant follicle-stimulating hormone (rFSH) followed by intrauterine insemination (IUI). STUDY DESIGN: This was a multicenter, prospective, randomized, open-label, assessor-blind, controlled trial of females (n = 54), ages 18 to 39 undergoing superovulation/IUI with up to 4 cycles of superovulation/IUI without Ganirelix (n = 66), or up to 4 cycles of superovulation/IUI with the addition of Ganirelix (n = 52). RESULTS: No statistically significant difference in clinical pregnancy rates per cycle initiated was found for patients in the treatment or control group (12% vs 7%, P =.29). Other variables assessed, including endometrial thickness, size of follicles, peak serum estradiol levels, mid-lutea progesterone levels, and total vials of rFSH used also showed no statistically significant difference. CONCLUSION: Superovulation/IUI cycles using Ganirelix produce similar pregnancy rates when compared with cycles not using a GnRH antagonist, although there is a trend towards better pregnancy rates in cycles with Ganirelix. PMID- 15343258 TI - Completion hysterectomy after radiation therapy for bulky cervical cancer stages IB, IIA, and IIB: complications and survival rates. AB - OBJECTIVE: The purpose of this study was to assess survival and morbidity when completion hysterectomy follows radiation for bulky cervical cancer. STUDY DESIGN: This was a retrospective observational descriptive review that assessed the survival and morbidity of patients with bulky cervical cancer that was treated with radiation followed by completion hysterectomy between 1993 and 2002. Chemotherapy, external beam radiation, and brachytherapy data were collected. RESULTS: Fifty-five cases were reviewed. Fifty-three patients received brachytherapy. Twenty-nine patients underwent sensitizing chemotherapy. All patients had hysterectomies. There were 12 early postoperative complications (21.8%) and 10 late complications (19.6%). Eleven patients are dead of disease (21.6%); 3 patients are alive with disease (5.9%), and 37 patients are free of disease (72.5%). Four patients were lost to follow-up. Seven patients who are free of disease had residual cancer in the specimen at hysterectomy. CONCLUSION: Complications of combined therapy were comparable to radiation or radical hysterectomy alone. In cases in which an incomplete response to radiation and chemotherapy leaves potential residual carcinoma, adjuvant hysterectomy may be a reasonable treatment option. PMID- 15343260 TI - Amniotic fluid index vs single deepest pocket technique during modified biophysical profile: a randomized clinical trial. AB - OBJECTIVE: The purpose of this study was to determine the superior technique, if either, of the amniotic fluid index (AFI) vs the single deepest pocket technique in predicting an adverse pregnancy outcome among high-risk patients undergoing antenatal testing. STUDY DESIGN: Patients having modified biophysical profile (nonstress test plus sonographic estimation of amniotic fluid) were randomized to either have AFI or determination of the presence or absence of a 2 x 1-cm single deepest pocket. RESULTS: Between January of 1997 and December of 2001, 1080 women were randomized with 530 women in the AFI arm, and 558 in the 2 x 1 pocket arm. The maternal demographics and prenatal complications were similar between groups. Significantly more patients were identified as having oligohydramnios using AFI (17%) compared with using 2 x 1 pocket (10%) ( P =.002). The overall rate of cesarean section for nonreassuring fetal heart rate (FHR) tracing was 3.8% (30 cases, with 16 cases in the AFI-monitored, and 14 cases in the 2 x 1 pocket monitored groups, respectively, P =.608). Logistic regression analysis showed no difference between the groups with respect to the ability to identify patients who underwent cesarean section for nonreassuring FHR tracing during labor ( P =.999). The umbilical artery pH <7.1 ( P =.688) and admission to the newborn intensive care unit were also comparable between groups. CONCLUSION: During antepartum fetal surveillance, use of single deepest pocket compared with amniotic fluid index is associated with a significantly lower rate of suspected oligohydramnios. PMID- 15343262 TI - Obesity: physiologic changes and challenges during laparoscopy. AB - OBJECTIVE: The purpose of this report was to describe the evidence for the benefits of laparoscopic surgery in obese patients, to review the physiologic abnormalities that are associated with obesity, and to explore surgical techniques that will enable surgeons to perform laparoscopy successfully in obese patients. STUDY DESIGN: This article is a review of the available literature on obesity, the physiologic changes that occur in obese patients, and the impact of these changes on laparoscopy outcomes. RESULTS: Obesity is associated with sudden death and a wide range of morbid conditions such as hypertension, atherosclerosis, angina, chronic obstructive disease, and diabetes mellitus. Recent studies have demonstrated that obese patients who undergo laparoscopy have shorter hospital stays, less postoperative pain, and fewer wound infections when compared with obese patients who undergo laparotomy. Laparoscopy patients also have fewer postoperative ileus and fevers. CONCLUSION: With thorough preparation and careful preoperative evaluation, laparoscopy can be performed safely and is the preferred surgical method in obese patients. PMID- 15343263 TI - Low selenium is associated with the occurrence of preeclampsia in women from the United Kingdom. PMID- 15343265 TI - HLA-DRB1, TAP2/TAP1, and HLA-DPB1 haplotypes in Finnish juvenile idiopathic arthritis: more complexity within the MHC. AB - This study further defines genetic susceptibility to JIA in the region centromeric to HLA-DRB1. DNA from 234 Finnish JIA nuclear families and 639 elderly Finnish control individuals was genotyped for five functional SNPs within the TAP2 and TAP1 loci ( approximately 200 kb centromeric of HLA-DRB1). Subsets of the controls (186) and patients (145) that had been previously typed for HLA DRB1 were also genotyped by sequence for the HLA-DPB1 locus. Case/control and transmission disequilibrium test (TDT) methods revealed an association with the DPB1(*)030101 allele for JIA (OR 2.3, 95% CI 1.5-3.5). Notably, a detailed haplotypic analysis of the TAP2/TAP1 loci and their interaction with the HLA DPB1(*)030101 and DRB1(*)08 and (*)11 alleles showed a variety of over represented and under-represented TAP2/TAP1 haplotypes not evident in the single marker analysis. The strongest effect was observed in the polyarticular RF negative JIA subgroup for the 2-2-1-2-1 TAP2/TAP1 haplotype (TAP2B and TAP1A alleles) which showed an independent effect from both DRB1(*)08 and (*)11 (P<0.000003) and DPB1(*)030101 (P=0.02). We have provided evidence that the extended haplotypes (including HLA-DRB1, TAP2/TAP1, and HLA-DPB1) of pauciarticular and polyarticular RF negative disease are distinct. This observation may have implications for functional etiological differences between the pauciarticular and polyarticular JIA patients. PMID- 15343266 TI - Defective apoptosis and B-cell lymphomas in mice with p53 point mutation at Ser 23. AB - Phosphorylation of the p53 tumor suppressor at Ser20 (murine Ser23) has been proposed to be critical for disrupting p53 interaction with its negative regulator, MDM2, and allowing p53 stabilization. To determine the importance of Ser23 for the function of p53 in vivo, we generated a mouse in which the endogenous p53 locus was targeted to replace Ser23 with alanine. We show that, in mouse embryonic fibroblasts generated from Ser23 mutant mice, Ser23 mutation did not dramatically reduce IR-induced p53 protein stabilization or p53-dependent cell cycle arrest. However, in Ser23 mutant thymocytes and in the developing cerebellum, p53 stabilization following IR was decreased and resistance to apoptosis was observed. Homozygous Ser23 mutant animals had a reduced lifespan, but did not develop thymic lymphomas or sarcomas that are characteristic of p53-/ mice. Instead, Ser23 mutant animals died between 1 and 2 years with tumors that were most commonly of B-cell lineage. These data support an important role for Ser20/23 phosphorylation in p53 stabilization, apoptosis and tumor suppression. PMID- 15343267 TI - Lats2/Kpm is required for embryonic development, proliferation control and genomic integrity. AB - The Drosophila melanogaster warts/lats tumour suppressor has two mammalian counterparts LATS1/Warts-1 and LATS2/Kpm. Here, we show that mammalian Lats orthologues exhibit distinct expression profiles according to germ cell layer origin. Lats2(-/-) embryos show overgrowth in restricted tissues of mesodermal lineage; however, lethality ultimately ensues on or before embryonic day 12.5 preceded by defective proliferation. Lats2(-/-) mouse embryonic fibroblasts (MEFs) acquire growth advantages and display a profound defect in contact inhibition of growth, yet exhibit defective cytokinesis. Lats2(-/-) embryos and MEFs display centrosome amplification and genomic instability. Lats2 localizes to centrosomes and overexpression of Lats2 suppresses centrosome overduplication induced in wild-type MEFs and reverses centrosome amplification inherent in Lats2(-/-) MEFs. These findings indicate an essential role of Lats2 in the integrity of processes that govern centrosome duplication, maintenance of mitotic fidelity and genomic stability. PMID- 15343268 TI - Tandem LIM domains provide synergistic binding in the LMO4:Ldb1 complex. AB - Nuclear LIM-only (LMO) and LIM-homeodomain (LIM-HD) proteins have important roles in cell fate determination, organ development and oncogenesis. These proteins contain tandemly arrayed LIM domains that bind the LIM interaction domain (LID) of the nuclear adaptor protein LIM domain-binding protein-1 (Ldb1). We have determined a high-resolution X-ray crystal structure of LMO4, a putative breast oncoprotein, in complex with Ldb1-LID, providing the first example of a tandem LIM:Ldb1-LID complex and the first structure of a type-B LIM domain. The complex possesses a highly modular structure with Ldb1-LID binding in an extended manner across both LIM domains of LMO4. The interface contains extensive hydrophobic and electrostatic interactions and multiple backbone-backbone hydrogen bonds. A mutagenic screen of Ldb1-LID, assessed by yeast two-hybrid and competition ELISA analysis, identified key features at the interface and revealed that the interaction is tolerant to mutation. These combined properties provide a mechanism for the binding of Ldb1 to numerous LMO and LIM-HD proteins. Furthermore, the modular extended interface may form a general mode of binding to tandem LIM domains. PMID- 15343269 TI - Transcriptional complexes engaged by apo-estrogen receptor-alpha isoforms have divergent outcomes. AB - Unliganded (apo-) estrogen receptor alpha (ERalpha, NR3A1) is classically considered as transcriptionally unproductive. Reassessing this paradigm demonstrated that apo-human ERalpha (ERalpha66) and its N-terminally truncated isoform (ERalpha46) are both predominantly nuclear transcription factors that cycle on the endogenous estrogen-responsive pS2 gene promoter in vivo. Importantly, isoform-specific consequences occur in terms of poising the promoter for transcription, as evaluated by determining (i) the engagement of several cofactors and the resulting nucleosomal organization; and (ii) the CpG methylation state of the pS2 promoter. Although transcriptionally unproductive, cycling of apo-ERalpha66 prepares the promoter to respond to ligand, through sequentially targeting chromatin remodeling complexes and general transcription factors. Additionally, apo-ERalpha46 recruits corepressors, following engagement of cofactors identical to those recruited by apo-ERalpha66. Together, these data describe differential activities of ERalpha isoforms. Furthermore, they depict the maintenance of a promoter in a repressed state as a cyclical process that is intrinsically dependent on initial poising of the promoter. PMID- 15343270 TI - Protocadherin FAT1 binds Ena/VASP proteins and is necessary for actin dynamics and cell polarization. AB - Cell migration requires integration of cellular processes resulting in cell polarization and actin dynamics. Previous work using tools of Drosophila genetics suggested that protocadherin fat serves in a pathway necessary for determining cell polarity in the plane of a tissue. Here we identify mammalian FAT1 as a proximal element of a signaling pathway that determines both cellular polarity in the plane of the monolayer and directed actin-dependent cell motility. FAT1 is localized to the leading edge of lamellipodia, filopodia, and microspike tips where FAT1 directly interacts with Ena/VASP proteins that regulate the actin polymerization complex. When targeted to mitochondrial outer leaflets, FAT1 cytoplasmic domain recruits components of the actin polymerization machinery sufficient to induce ectopic actin polymerization. In an epithelial cell wound model, FAT1 knockdown decreased recruitment of endogenous VASP to the leading edge and resulted in impairment of lamellipodial dynamics, failure of polarization, and an attenuation of cell migration. FAT1 may play an integrative role regulating cell migration by participating in Ena/VASP-dependent regulation of cytoskeletal dynamics at the leading edge and by transducing an Ena/VASP independent polarity cue. PMID- 15343271 TI - The polarity-inducing kinase Par-1 controls Xenopus gastrulation in cooperation with 14-3-3 and aPKC. AB - Par (partitioning-defective) genes were originally identified in Caenorhabditis elegans as determinants of anterior/posterior polarity. However, neither their function in vertebrate development nor their action mechanism has been fully addressed. Here we show that two members of Par proteins, 14-3-3 (Par-5) and atypical PKC (aPKC), regulate the serine/threonine kinase Par-1 to control Xenopus gastrulation. We find first that Xenopus Par-1 (xPar-1) is essential for gastrulation but not for cell fate specification during early embryonic development. We then find that xPar-1 binds to 14-3-3 in an aPKC-dependent manner. Our analyses identify two aPKC phosphorylation sites in xPar-1, which are essential for 14-3-3 binding and for proper gastrulation movements. The aPKC phosphorylation-dependent binding of xPar-1 to 14-3-3 does not markedly affect the kinase activity of xPar-1, but induces relocation of xPar-1 from the plasma membranes to the cytoplasm. Finally, we show that Xenopus aPKC and its binding partner Xenopus Par-6 are also essential for gastrulation. Thus, our results identify a requirement of Par proteins for Xenopus gastrulation and reveal a novel interrelationship within Par proteins that may provide a general mechanism for spatial control of Par-1. PMID- 15343272 TI - A single-centre assessment of long-term quality-of-life status after sibling allogeneic stem cell transplantation for chronic myeloid leukaemia in first chronic phase. AB - A total of 75 patients underwent sibling allogeneic stem cell transplantation (SCT) for chronic myeloid leukaemia in first chronic phase from 1984 to 2000. Of these patients, 51 (68%) were alive at a median follow-up of 98 months (range 34 217 months). Nine (18%) patients relapsed and seven (14%) received donor lymphocyte transfusions. Quality of life (QoL) was assessed cross-sectionally using the EORTC QLQ-C30, a Leukaemia-BMT-specific module and questionnaires on sexual functioning, fertility and late effects. A total of 46 (90%) replied. Scores for Role (P=0.018) and Cognitive (P<0.001) function were significantly lower when compared to an age-adjusted general population. Dyspnoea (P=0.022) and Financial Difficulties (P<0.001) were significantly more common in the SCT group. No difference was found for scores in the Physical, Emotional and Social domains or the overall Global Health Status/QoL. Decreased sexual functioning was found in one-third of respondents. Although most BMT recipients reported a good QoL, a minority have difficulty with reintegration into professional roles and consequent monetary problems. Identified cognitive and sexual impairments highlight the need for long-term access to psychosocial support. PMID- 15343273 TI - BRCA1 and BRCA2: 1994 and beyond. AB - The discovery of the first gene associated with hereditary breast cancer, BRCA1, was anticipated to greatly increase our understanding of both hereditary and sporadic forms of breast cancer, and to lead to therapeutic and preventive breakthroughs. Much has been learned during the past decade about the genetic epidemiology of breast cancer, the ethnic distribution and clinical consequences of BRCA1 and BRCA2 mutations, and the central role of DNA repair in breast cancer susceptibility. The ability to translate this knowledge into novel treatments, however, remains elusive. PMID- 15343274 TI - Nuclear structure in cancer cells. AB - Nuclear architecture - the spatial arrangement of chromosomes and other nuclear components - provides a framework for organizing and regulating the diverse functional processes within the nucleus. There are characteristic differences in the nuclear architectures of cancer cells, compared with normal cells, and some anticancer treatments restore normal nuclear structure and function. Advances in understanding nuclear structure have revealed insights into the process of malignant transformation and provide a basis for the development of new diagnostic tools and therapeutics. PMID- 15343275 TI - Oncogenic mechanisms of the Helicobacter pylori CagA protein. AB - Infection with strains of Helicobacter pylori that carry the cytotoxin-associated antigen A (cagA) gene is associated with gastric carcinoma. Recent studies have shed light on the mechanism through which the cagA gene product, CagA, elicits pathophysiological actions. CagA is delivered into gastric epithelial cells by the bacterial type IV secretion system, where it deregulates the SHP2 oncoprotein. Intriguingly, CagA is noted for its variation, particularly at the SHP2-binding site, which could affect the potential of different strains of H. pylori to promote gastric carcinogenesis. PMID- 15343276 TI - Gallbladder cancer: lessons from a rare tumour. AB - Gallbladder cancer is a relatively rare form of malignancy about which our knowledge is scant. However, a unique combination of predisposing factors - including genetic predisposition, geographic distribution, female gender bias, chronic inflammation and congenital developmental abnormalities - makes this type of cancer unique and offers potential for understanding cancer pathogenesis in general. An understanding of how these risk factors contribute to the molecular basis of the disease is essential for understanding the origins of this unusual cancer. PMID- 15343277 TI - Gene-promoter hypermethylation as a biomarker in lung cancer. AB - Silencing of genes by aberrant promoter hypermethylation is now recognized as a crucial component in cancer initiation and progression. Highly sensitive assays have been developed to assess gene-promoter methylation in biological fluids. The detection of methylated genes in sputum could lead to the development of a screening test to non-invasively identify early cancer in high-risk people. PMID- 15343278 TI - Switching on kinases: oncogenic activation of BRAF and the PDGFR family. AB - The cytoplasmic serine/threonine kinase BRAF and receptor tyrosine kinases of the platelet-derived growth factor receptor (PDGFR) family are frequently activated in cancer by mutations of an equivalent amino acid. Structural studies have provided important insights into why these very different kinases share similar oncogenic hot spots and why the PDGFR juxtamembrane region is also a frequent oncogenic target. This research has implications for other kinases that are mutated in human tumours and for the treatment of cancer using kinase inhibitors. PMID- 15343279 TI - Transcription - guarding the genome by sensing DNA damage. AB - Cells induce the expression of DNA-repair enzymes, activate cell-cycle checkpoints and, under some circumstances, undergo apoptosis in response to DNA damaging agents. The mechanisms by which these cellular responses are triggered are not well understood, but there is recent evidence that the transcription machinery might be used in DNA-damage surveillance and in triggering DNA-damage responses to suppress mutagenesis. Transcription might also act as a DNA-damage dosimeter where the severity of blockage determines whether or not to induce cell death. Could transcription therefore be a potential therapeutic target for anticancer strategies? PMID- 15343280 TI - Radiation oncology: a century of achievements. AB - Over the twentieth century the discipline of radiation oncology has developed from an experimental application of X-rays to a highly sophisticated treatment of cancer. Experts from many disciplines - chiefly clinicians, physicists and biologists - have contributed to these advances. Whereas the emphasis in the past was on refining techniques to ensure the accurate delivery of radiation, the future of radiation oncology lies in exploiting the genetics or the microenvironment of the tumour to turn cancer from an acute disease to a chronic disease that can be treated effectively with radiation. PMID- 15343282 TI - What does 'harder than diamond' mean? PMID- 15343283 TI - Food for thought. AB - Food is a fascinating and important material that requires study of the same intellectual rigour as that performed for the more usual substances reported within materials science journals. A look at some basic foodstuffs helps to identify why, and also why the science involved is intrinsically exciting. PMID- 15343285 TI - Friction: terms of detachment. PMID- 15343286 TI - Biomolecular self-assembly: stacks of viruses. PMID- 15343287 TI - Polymers: a multitude of macromolecules. PMID- 15343288 TI - Light-emitting devices: turning the tables on surface plasmons. PMID- 15343289 TI - Biomimetic membrane modelling: pictures from the twilight zone. PMID- 15343291 TI - Diamond-structured photonic crystals. AB - Certain periodic dielectric structures can prohibit the propagation of light for all directions within a frequency range. These 'photonic crystals' allow researchers to modify the interaction between electromagnetic fields and dielectric media from radio to optical wavelengths. Their technological potential, such as the inhibition of spontaneous emission, enhancement of semiconductor lasers, and integration and miniaturization of optical components, makes the search for an easy-to-craft photonic crystal with a large bandgap a major field of study. This progress article surveys a collection of robust complete three-dimensional dielectric photonic-bandgap structures for the visible and near-infrared regimes based on the diamond morphology together with their specific fabrication techniques. The basic origin of the complete photonic bandgap for the 'champion' diamond morphology is described in terms of dielectric modulations along principal directions. Progress in three-dimensional interference lithography for fabrication of near-champion diamond-based structures is also discussed. PMID- 15343292 TI - Not just academic. PMID- 15343293 TI - Distributing the costs of climate change. PMID- 15343294 TI - Ethics review slams government panels over conflicts of interest. PMID- 15343295 TI - Disaster movie highlights transatlantic divide. PMID- 15343296 TI - Organizers claim success for Stockholm science jamboree. PMID- 15343297 TI - Wildlife campaigners fight planned site for Scripps Florida. PMID- 15343298 TI - Lab chiefs fear European rules will cost postdoc jobs. PMID- 15343299 TI - Thailand faces dilemma over bird flu vaccine. PMID- 15343300 TI - Microchip industry proposes broad survey of worker health. PMID- 15343301 TI - Five-year grant gets bird database off to a flying start. PMID- 15343303 TI - Climate change: crunch time for Kyoto. PMID- 15343304 TI - Physics and music: Brothers in art. PMID- 15343305 TI - Need for economists to set global priorities. PMID- 15343306 TI - Biosecurity must be internationally supervised. PMID- 15343307 TI - Linnean Society backs Godfray on use of web. PMID- 15343308 TI - Politics, morals and embryos. PMID- 15343313 TI - Midsummer madness. PMID- 15343314 TI - Astrobiology: message in a bottle. PMID- 15343315 TI - Evolutionary biology: time, space and genomes. PMID- 15343316 TI - Statistical physics: hear the noise. PMID- 15343317 TI - Cell biology: regulated self-cannibalism. PMID- 15343318 TI - Planet formation: the core problem. PMID- 15343319 TI - Theoretical biology: mushrooms in cyberspace. PMID- 15343320 TI - Cancer: cell survival guide. PMID- 15343321 TI - Astronomy: the quiet one. PMID- 15343324 TI - Animal behaviour: use of dung as a tool by burrowing owls. AB - Reports of tool usage by birds tend to be anecdotal as only a few individuals may be involved and the behaviour observed can often be interpreted in other ways. Here we describe the widespread collection of mammalian dung by burrowing owls (Athene cunicularia) and show that they use this dung as a bait to attract dung beetles, a major item of prey. Our controlled investigation provides an unambiguous estimate of the importance of tool use in a wild animal. PMID- 15343325 TI - Botany: a new self-pollination mechanism. AB - Pollen grains from most flowering plants are transported by wind or animals and deposited on the receptive surface of the stigma of a different individual, but self-pollination is also common. We have discovered a new process for self pollination in the laterally orientated flowers of a Chinese herb, in which a film of pollen is transported from the anther (pollen sacs) by an oily emulsion that slides sideways along the flower's style and into the individual's own stigma. This mode of self-pollination is a new addition to the broad range of genetic and morphological mechanisms that have evolved in flowering plants (angiosperms), and may be common in species growing in shady, windless and insect poor habitats. PMID- 15343326 TI - Active foundering of a continental arc root beneath the southern Sierra Nevada in California. AB - Seismic data provide images of crust-mantle interactions during ongoing removal of the dense batholithic root beneath the southern Sierra Nevada mountains in California. The removal appears to have initiated between 10 and 3 Myr ago with a Rayleigh-Taylor-type instability, but with a pronounced asymmetric flow into a mantle downwelling (drip) beneath the adjacent Great Valley. A nearly horizontal shear zone accommodated the detachment of the ultramafic root from its granitoid batholith. With continuing flow into the mantle drip, viscous drag at the base of the remaining approximately 35-km-thick crust has thickened the crust by approximately 7 km in a narrow welt beneath the western flank of the range. Adjacent to the welt and at the top of the drip, a V-shaped cone of crust is being dragged down tens of kilometres into the core of the mantle drip, causing the disappearance of the Moho in the seismic images. Viscous coupling between the crust and mantle is therefore apparently driving present-day surface subsidence. PMID- 15343327 TI - Inscribed matter as an energy-efficient means of communication with an extraterrestrial civilization. AB - It is well known that electromagnetic radiation-radio waves-can in principle be used to communicate over interstellar distances. By contrast, sending physical artefacts has seemed extravagantly wasteful of energy, and imagining human travel between the stars even more so. The key consideration in earlier work, however, was the perceived need for haste. If extraterrestrial civilizations existed within a few tens of light years, radio could be used for two-way communication on timescales comparable to human lifetimes (or at least the longevities of human institutions). Here we show that if haste is unimportant, sending messages inscribed on some material can be strikingly more energy efficient than communicating by electromagnetic waves. Because messages require protection from cosmic radiation and small messages could be difficult to find among the material clutter near a recipient, 'inscribed matter' is most effective for long archival messages (as opposed to potentially short "we exist" announcements). The results suggest that our initial contact with extraterrestrial civilizations may be more likely to occur through physical artefacts-essentially messages in a bottle-than via electromagnetic communication. PMID- 15343328 TI - Spectroscopy of spontaneous spin noise as a probe of spin dynamics and magnetic resonance. AB - Not all noise in experimental measurements is unwelcome. Certain fundamental noise sources contain valuable information about the system itself-a notable example being the inherent voltage fluctuations (Johnson noise) that exist across any resistor, which allow the temperature to be determined. In magnetic systems, fundamental noise can exist in the form of random spin fluctuations. For example, statistical fluctuations of N paramagnetic spins should generate measurable noise of order N spins, even in zero magnetic field. Here we exploit this effect to perform perturbation-free magnetic resonance. We use off-resonant Faraday rotation to passively detect the magnetization noise in an equilibrium ensemble of paramagnetic alkali atoms; the random fluctuations generate spontaneous spin coherences that precess and decay with the same characteristic energy and timescales as the macroscopic magnetization of an intentionally polarized or driven ensemble. Correlation spectra of the measured spin noise reveal g-factors, nuclear spin, isotope abundance ratios, hyperfine splittings, nuclear moments and spin coherence lifetimes-without having to excite, optically pump or otherwise drive the system away from thermal equilibrium. These noise signatures scale inversely with interaction volume, suggesting a possible route towards non perturbative, sourceless magnetic resonance of small systems. PMID- 15343329 TI - Negative intrinsic resistivity of an individual domain wall in epitaxial (Ga,Mn)As microdevices. AB - Magnetic domains, and the boundaries that separate them (domain walls, DWs), play a central role in the science of magnetism. Understanding and controlling domains is important for many technological applications in spintronics, and may lead to new devices. Although theoretical efforts have elucidated several mechanisms underlying the resistance of a single DW, various experiments report conflicting results, even for the overall sign of the DW resistance. The question of whether an individual DW gives rise to an increase or decrease of the resistance therefore remains open. Here we report an approach to DW studies in a class of ferromagnetic semiconductors (as opposed to metals) that offer promise for spintronics. These experiments involve microdevices patterned from monocrystalline (Ga,Mn)As epitaxial layers. The giant planar Hall effect that we previously observed in this material enables direct, real-time observation of the propagation of an individual magnetic DW along multiprobe devices. We apply steady and pulsed magnetic fields, to trap and carefully position an individual DW within each separate device studied. This protocol reproducibly enables high resolution magnetoresistance measurements across an individual wall. We consistently observe negative intrinsic DW resistance that scales with channel width. This appears to originate from sizeable quantum corrections to the magnetoresistance. PMID- 15343330 TI - Decline of surface temperature and salinity in the western tropical Pacific Ocean in the Holocene epoch. AB - In the present-day climate, surface water salinities are low in the western tropical Pacific Ocean and increase towards the eastern part of the basin. The salinity of surface waters in the tropical Pacific Ocean is thought to be controlled by a combination of atmospheric convection, precipitation, evaporation and ocean dynamics, and on interannual timescales significant variability is associated with the El Nino/Southern Oscillation cycles. However, little is known about the variability of the coupled ocean-atmosphere system on timescales of centuries to millennia. Here we combine oxygen isotope and Mg/Ca data from foraminifers retrieved from three sediment cores in the western tropical Pacific Ocean to reconstruct Holocene sea surface temperatures and salinities in the region. We find a decrease in sea surface temperatures of approximately 0.5 degrees C over the past 10,000 yr, whereas sea surface salinities decreased by approximately 1.5 practical salinity units. Our data imply either that the Pacific basin as a whole has become progressively less salty or that the present salinity gradient along the Equator has developed relatively recently. PMID- 15343331 TI - Osmium isotopic constraints on the nature of the DUPAL anomaly from Indian mid ocean-ridge basalts. AB - The isotopic compositions of mid-ocean-ridge basalts (MORB) from the Indian Ocean have led to the identification of a large-scale isotopic anomaly relative to Pacific and Atlantic ocean MORB. Constraining the origin of this so-called DUPAL anomaly may lead to a better understanding of the genesis of upper-mantle heterogeneity. Previous isotopic studies have proposed recycling of ancient subcontinental lithospheric mantle or sediments with oceanic crust to be responsible for the DUPAL signature. Here we report Os, Pb, Sr and Nd isotopic compositions of Indian MORB from the Central Indian ridge, the Rodriguez triple junction and the South West Indian ridge. All measured samples have higher (187)Os/(188)Os ratios than the depleted upper-mantle value and Pb, Sr and Nd isotopic compositions that imply the involvement of at least two distinct enriched components in the Indian upper-mantle. Using isotopic and geodynamical arguments, we reject both subcontinental lithospheric mantle and recycled sediments with oceanic crust as the cause of the DUPAL anomaly. Instead, we argue that delamination of lower continental crust may explain the DUPAL isotopic signature of Indian MORB. PMID- 15343332 TI - The evolution of mullerian mimicry in multispecies communities. AB - Prey species that are unprofitable to attack often share conspicuous colours and patterns with other coexisting defended species. This phenomenon, termed mullerian mimicry, has long been explained as a consequence of selection on defended prey to adopt a common way of advertising their unprofitability. However, studies using two unpalatable prey types have not always supported this theory. Here we show, using a system of humans hunting for computer-generated prey, that predators do not always generate strong selection for mimicry when there are two unprofitable prey types. By contrast, we demonstrate that when predators are faced with a range of different prey species, selection on unprofitable prey to resemble one another can be intense. Here the primary selective force is not one in which predators evaluate the profitabilities of distinct prey types independently, but one in which predators learn better to avoid unprofitable phenotypes that share traits distinguishing them from profitable prey. This need to simplify decision making readily facilitates the spread of imperfect mimetic forms from rarity, and suggests that mullerian mimicry is more likely to arise in multispecies communities. PMID- 15343333 TI - Hox cluster disintegration with persistent anteroposterior order of expression in Oikopleura dioica. AB - Tunicate embryos and larvae have small cell numbers and simple anatomical features in comparison with other chordates, including vertebrates. Although they branch near the base of chordate phylogenetic trees, their degree of divergence from the common chordate ancestor remains difficult to evaluate. Here we show that the tunicate Oikopleura dioica has a complement of nine Hox genes in which all central genes are lacking but a full vertebrate-like set of posterior genes is present. In contrast to all bilaterians studied so far, Hox genes are not clustered in the Oikopleura genome. Their expression occurs mostly in the tail, with some tissue preference, and a strong partition of expression domains in the nerve cord, in the notochord and in the muscle. In each tissue of the tail, the anteroposterior order of Hox gene expression evokes spatial collinearity, with several alterations. We propose a relationship between the Hox cluster breakdown, the separation of Hox expression domains, and a transition to a determinative mode of development. PMID- 15343334 TI - Biological abnormality of impaired reading is constrained by culture. AB - Developmental dyslexia is characterized by a severe reading problem in people who have normal intelligence and schooling. Impaired reading of alphabetic scripts is associated with dysfunction of left temporoparietal brain regions. These regions perform phonemic analysis and conversion of written symbols to phonological units of speech (grapheme-to-phoneme conversion); two central cognitive processes that mediate reading acquisition. Furthermore, it has been assumed that, in contrast to cultural diversities, dyslexia in different languages has a universal biological origin. Here we show using functional magnetic resonance imaging with reading-impaired Chinese children and associated controls, that functional disruption of the left middle frontal gyrus is associated with impaired reading of the Chinese language (a logographic rather than alphabetic writing system). Reading impairment in Chinese is manifested by two deficits: one relating to the conversion of graphic form (orthography) to syllable, and the other concerning orthography-to-semantics mapping. Both of these processes are critically mediated by the left middle frontal gyrus, which functions as a centre for fluent Chinese reading that coordinates and integrates various information about written characters in verbal and spatial working memory. This finding provides an insight into the fundamental pathophysiology of dyslexia by suggesting that rather than having a universal origin, the biological abnormality of impaired reading is dependent on culture. PMID- 15343335 TI - Netrin-1 controls colorectal tumorigenesis by regulating apoptosis. AB - The expression of the protein DCC (deleted in colorectal cancer) is lost or markedly reduced in numerous cancers and in the majority of colorectal cancers due to loss of heterozygosity in chromosome 18q, and has therefore been proposed to be a tumour suppressor. However, the rarity of mutations found in DCC, the lack of cancer predisposition of DCC mutant mice, and the presence of other tumour suppressor genes in 18q have raised doubts about the function of DCC as a tumour suppressor. Unlike classical tumour suppressors, DCC has been shown to induce apoptosis conditionally: by functioning as a dependence receptor, DCC induces apoptosis unless DCC is engaged by its ligand, netrin-1 (ref. 3). Here we show that inhibition of cell death by enforced expression of netrin-1 in mouse gastrointestinal tract leads to the spontaneous formation of hyperplastic and neoplastic lesions. Moreover, in the adenomatous polyposis coli mutant background associated with adenoma formation, enforced expression of netrin-1 engenders aggressive adenocarcinomatous malignancies. These data demonstrate that netrin-1 can promote intestinal tumour development, probably by regulating cell survival. Thus, a netrin-1 receptor or receptors function as conditional tumour suppressors. PMID- 15343336 TI - A glycolipid of hypervirulent tuberculosis strains that inhibits the innate immune response. AB - Fifty million new infections with Mycobacterium tuberculosis occur annually, claiming 2-3 million lives from tuberculosis worldwide. Despite the apparent lack of significant genetic heterogeneity between strains of M. tuberculosis, there is mounting evidence that considerable heterogeneity exists in molecules important in disease pathogenesis. These differences may manifest in the ability of some isolates to modify the host cellular immune response, thereby contributing to the observed diversity of clinical outcomes. Here we describe the identification and functional relevance of a highly biologically active lipid species-a polyketide synthase-derived phenolic glycolipid (PGL) produced by a subset of M. tuberculosis isolates belonging to the W-Beijing family that show 'hyperlethality' in murine disease models. Disruption of PGL synthesis results in loss of this hypervirulent phenotype without significantly affecting bacterial load during disease. Loss of PGL was found to correlate with an increase in the release of the pro-inflammatory cytokines tumour-necrosis factor-alpha and interleukins 6 and 12 in vitro. Furthermore, the overproduction of PGL by M. tuberculosis or the addition of purified PGL to monocyte-derived macrophages was found to inhibit the release of these pro-inflammatory mediators in a dose dependent manner. PMID- 15343337 TI - Involvement of targeted proteolysis in plant genetic transformation by Agrobacterium. AB - Genetic transformation of plant cells by Agrobacterium represents a unique case of trans-kingdom DNA transfer. During this process, Agrobacterium exports its transferred (T) DNA and several virulence (Vir) proteins into the host cell, within which T-DNA nuclear import is mediated by VirD2 (ref. 3) and VirE2 (ref. 4) and their host cell interactors AtKAP-alpha and VIP1 (ref. 6), whereas its integration is mediated mainly by host cell proteins. The factors involved in the uncoating of T-DNA from its cognate proteins, which occurs before integration into the host genome, are still unknown. Here, we report that VirF-one of the few known exported Vir proteins whose function in the host cell remains unknown-is involved in targeted proteolysis of VIP1 and VirE2. We show that VirF localizes to the plant cell nucleus and interacts with VIP1, a nuclear protein. VirF, which contains an F-box motif, significantly destabilizes both VIP1 and VirE2 in yeast cells. Destabilization of VIP1 in the presence of VirF was then confirmed in planta. These results suggest that VIP1 and its cognate VirE2 are specifically targeted by the VirF-containing Skp1-Cdc53-cullin-F-box complex for proteolysis. The critical role of proteasomal degradation in Agrobacterium-mediated genetic transformation was also evident from inhibition of T-DNA expression by a proteasomal inhibitor. PMID- 15343338 TI - Centrosomes direct cell polarity independently of microtubule assembly in C. elegans embryos. AB - Polarity establishment requires a symmetry-breaking event, resulting in an axis along which determinants are segregated. In Caenorhabditis elegans, oocytes are apolar and are triggered to polarize rapidly along one axis after fertilization. The establishment of this first polarity axis is revealed by the asymmetric distribution of PAR proteins and cortical activity in the one-celled embryo. Current evidence suggests that the centrosome-pronucleus complex contributed by the sperm is involved in defining the polarization axis. Here we directly assess the contribution of the centrosome to polarity establishment by laser ablating the centrosome before and during polarization. We find that the centrosome is required to initiate polarity but not to maintain it. Initiation of polarity coincides with the proximity of the centrosome to the cortex and the assembly of pericentriolar material on the immature sperm centrosome. Depletion of microtubules or the microtubule nucleator gamma-tubulin did not affect polarity establishment. These results demonstrate that the centrosome provides an initiating signal that polarizes C. elegans embryos and indicate that this signalling event might be independent of the role of the centrosome as a microtubule nucleator. PMID- 15343339 TI - Transcriptional regulatory code of a eukaryotic genome. AB - DNA-binding transcriptional regulators interpret the genome's regulatory code by binding to specific sequences to induce or repress gene expression. Comparative genomics has recently been used to identify potential cis-regulatory sequences within the yeast genome on the basis of phylogenetic conservation, but this information alone does not reveal if or when transcriptional regulators occupy these binding sites. We have constructed an initial map of yeast's transcriptional regulatory code by identifying the sequence elements that are bound by regulators under various conditions and that are conserved among Saccharomyces species. The organization of regulatory elements in promoters and the environment-dependent use of these elements by regulators are discussed. We find that environment-specific use of regulatory elements predicts mechanistic models for the function of a large population of yeast's transcriptional regulators. PMID- 15343340 TI - Structure of the acrosomal bundle. AB - In the unactivated Limulus sperm, a 60- micro m-long bundle of actin filaments crosslinked by the protein scruin is bent and twisted into a coil around the base of the nucleus. At fertilization, the bundle uncoils and fully extends in five seconds to support a finger of membrane known as the acrosomal process. This biological spring is powered by stored elastic energy and does not require the action of motor proteins or actin polymerization. In a 9.5-A electron cryomicroscopic structure of the extended bundle, we show that twist, tilt and rotation of actin-scruin subunits deviate widely from a 'standard' F-actin filament. This variability in structural organization allows filaments to pack into a highly ordered and rigid bundle in the extended state and suggests a mechanism for storing and releasing energy between coiled and extended states without disassembly. PMID- 15343343 TI - The skills hunt. PMID- 15343344 TI - A distinctive nuclear morphology in acute myeloid leukemia is strongly associated with loss of HLA-DR expression and FLT3 internal tandem duplication. AB - In a 5-year survey of nonpromyelocytic/nonmonocytic acute myeloid leukemias (AMLs) diagnosed in the University of Washington Hematopathology Laboratory, we identified 19 cases containing distinctive, cup-like nuclear indentation in 10% or more of the blasts ('AML-cuplike'). Fourteen of these cases (74%) demonstrated near-complete loss of HLA-DR expression, while the other five cases showed partial loss of HLA-DR. A total of 16 of the cases (84%) demonstrated internal tandem duplication (ITD) of the Flt3 gene. When compared to a selected set of AMLs lacking this nuclear morphology, AML-cuplike was significantly more likely to lack HLA-DR and CD34 expression, to express CD123 without CD133, to have a normal karyotype, and to harbor the Flt3 ITD. To characterize AML-cuplike in an unselected series of AMLs, we analyzed 42 consecutive nonpromyelocytic/nonmonocytic AMLs diagnosed in our laboratory during a 6-month period in 2002. Strikingly, in this unselected series, there was a statistically significant coincidence of invaginated nuclear morphology, loss of HLA-DR, and presence of the Flt3 ITD beyond that expected if these three features were unrelated, suggesting that AMLs with these three features may represent a distinct AML subset. PMID- 15343345 TI - Antitumor effects of bortezomib (PS-341) on primary effusion lymphomas. AB - Primary effusion lymphomas (PELs) are a rare type of non-Hodgkin's lymphoma that are resistant to cytotoxic chemotherapy. PELs manifest constitutive activation of nuclear factor kappa B (NF-kappaB), and inhibition of NF-kappaB induces apoptosis of PELs and sensitizes to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced death. Bortezomib (PS-341), a peptidyl boronic acid inhibitor of the proteasome, is a potent agent against a wide range of hematologic malignancies and has been shown to inhibit NF-kappaB. Thus, we examined the cytotoxic effects of bortezomib alone and in combination with various drugs. Bortezomib potently inhibited NF-kappaB in PEL cells in a dose-dependent manner. In addition, bortezomib inhibited growth and induced apoptosis of PEL cell lines (IC(50) values of 3.4-5.0 nM). Results of drug interactions between bortezomib and chemotherapy (doxorubicin and Taxol) were schedule-dependent: synergistic interactions were generally observed when PEL cells were pretreated with bortezomib prior to chemotherapy, whereas additive or even antagonistic interactions occurred with chemotherapy pretreatment or simultaneous treatment with bortezomib and chemotherapy. Most schedules of bortezomib and dexamethasone were synergistic, although pretreatment with dexamethasone resulted in additive interactions. Effects of combinations of bortezomib and TRAIL were generally additive. Thus, bortezomib represents a promising potential therapy for the treatment of PEL. PMID- 15343346 TI - Distinct gene signatures of transient and acute megakaryoblastic leukemia in Down syndrome. AB - Approximately 10% of newborns with Down syndrome develop Transient Leukemia (TL), a disorder that is unique to infants with constitutional trisomy 21 (or trisomy 21 mosaicism). TL blasts disappear spontaneously within the first 3 months of life in the majority of cases. Despite the resolution of TL, 20-30% of these newborns will go on to develop acute megakaryoblastic leukemia (AMKL) later in life. In this study, samples from both TL and AMKL patients were examined using cDNA microarrays to study the pathogenic progression from TL to AMKL. TL and AMKL samples partition separately by cluster analysis, and AMKL samples had substantial increases in apolipoprotein C-I, transporter 1, myosin alkali light chain 4, and spermidine/spermine N-acetyltransferase, compared to TL samples. Although these findings will require validation in an independent series of TL and AMKL samples, they indicate that TL and AMKL have distinct gene signatures, and provide a basis for studies of the different mechanisms underlying either the resolution of TL or its progression to AMKL. PMID- 15343347 TI - Defective blood dendritic cells in chronic myeloid leukemia correlate with high plasmatic VEGF and are not normalized by imatinib mesylate. AB - Human blood dendritic cells (DC) comprise plasmacytoid DC (PDC) and myeloid DC (MDC), which both prime antitumor T-cell responses. We prospectively monitored blood DC in 30 chronic myeloid leukemia (CML) patients before and after imatinib mesylate therapy. We found a dramatic reduction in PDC and MDC prior treatment. This reduction was associated with high plasmatic vascular endothelial growth factor (VEGF), a central regulator of angiogenesis which also participates to tumor-associated immune deficiencies. Phenotypic analysis of DC revealed in some patients a deficient expression of BDCA-4/neuropilin-1 on PDC, a molecule involved in angiogenesis and DC-T-cell interactions. High VEGF correlated to an altered Th1/Th2 balance in vivo and shifted PDC-induced T-cell polarization towards Th2 in vitro. Upon imatinib treatment, plasmatic VEGF rapidly decreased and a normal BDCA-4 expression was restored. PDC and MDC increased but did not reach the levels observed in healthy individuals. We conclude that VEGF may be a key player in blood DC deficiency in CML and we show that imatinib inhibits VEGF overproduction. Incomplete recovery of blood DC under imatinib despite VEGF normalization suggests a negative impact of this drug on dendritopoiesis in vivo and may result in a sustained defect in DC-mediated anti-CML responses. PMID- 15343348 TI - Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation. AB - The clinically most suitable method for minimal residual disease (MRD) detection in chronic lymphocytic leukemia is still controversial. We prospectively compared MRD assessment in 158 blood samples of 74 patients with CLL after stem cell transplantation (SCT) using four-color flow cytometry (MRD flow) in parallel with consensus IgH-PCR and ASO IgH real-time PCR (ASO IgH RQ-PCR). In 25 out of 106 samples (23.6%) with a polyclonal consensus IgH-PCR pattern, MRD flow still detected CLL cells, proving higher sensitivity of flow cytometry over PCR genescanning with consensus IgH-primers. Of 92 samples, 14 (15.2%) analyzed in parallel by MRD flow and by ASO IgH RQ-PCR were negative by our flow cytometric assay but positive by PCR, thus demonstrating superior sensitivity of RQ-PCR with ASO primers. Quantitative MRD levels measured by both methods correlated well (r=0.93). MRD detection by flow and ASO IgH RQ-PCR were equally suitable to monitor MRD kinetics after allogeneic SCT, but the PCR method detected impending relapses after autologous SCT earlier. An analysis of factors that influence sensitivity and specificity of flow cytometry for MRD detection allowed to devise further improvements of this technique. PMID- 15343349 TI - CD38 modifications in chronic lymphocytic leukemia: are they relevant? PMID- 15343350 TI - Maternal Ramadan fasting and neonatal health. AB - This retrospective cohort study sought to determine the effects of maternal Ramadan fasting during pregnancy on neonatal birth weight as an important aspect of fetal health. It was carried out among healthy women who were admitted for their neonate delivery at two medical centers in Tehran from January to September 2000. Neonates of 284 mothers with a history of Ramadan fasting during pregnancy were compared with neonates of 255 mothers who did not have a history of fasting during their pregnancies. On univariate analysis, neonatal birth weight of the fasted group was 100 g more than those of the nonfasted group (p=0.009). However, body mass index (BMI) of the fasted mothers was greater than that of the nonfasted mothers. When controlling for maternal BMI on neonatal birth weight, multiple linear regression models showed that neonates of fasted women were 71 g heavier than those of the nonfasted group, which was not statistically significant (p=0.1). We conclude that maternal fasting during Ramadan did not have a significant effect on the neonatal birth weight. Other health effects that we did not observe could have occurred. PMID- 15343351 TI - Can oral sucrose reduce the pain and distress associated with screening for retinopathy of prematurity? AB - OBJECTIVE: Infants undergoing eye exams to screen for retinopathy of prematurity (ROP) demonstrate physiologic and behavioral manifestations of pain and distress. Oral sucrose has analgesic properties that might reduce these effects. AIM: To determine the efficacy of oral sucrose in reducing the pain/distress of eye exams for ROP. METHODS: A total of 32 infants about to undergo ROP screening exams received either oral sucrose [S] (N=16) or sterile water [C] (N=16) in a randomized, prospective and blinded fashion. Outcome measures included HR, RR, O(2) saturation, BP, pain (premature infant pain profile) and percent of time spent crying during the eye exam. RESULTS: The groups were similar in GA (weeks) (28+/-1.6), BW (kg) (1.04+/-0.26), postnatal age (days) 50.8+/-20.3, and study weight (kg) 1.88+/-0.40). Both groups demonstrated significant increases in HR, BP, and pain score in response to the exam. Infants in both groups spent the majority of time actively crying during the exam ([S] 53+/-35% vs [C] 63+/-31%. Infants receiving [S] showed a small but significant drop in O(2) saturation. No significant differences were seen between groups in physiologic or behavioral responses to the eye exam. CONCLUSION: Oral [S] was not effective in reducing pain/distress from the ROP screening exam. Alternative strategies should be considered to achieve adequate pain relief. PMID- 15343352 TI - Prevalence of hypertension, its awareness and control in adult population in Japan. PMID- 15343353 TI - Role of two angiotensinogen polymorphisms in blood pressure variation. AB - Genetic analysis of hypertension has yielded inconsistent results, making it difficult to draw clear conclusions regarding the impact of specific variants on blood pressure regulation. Among the most studied of the candidate genes for blood pressure regulation is angiotensinogen (AGT), but as with other candidate loci associations with blood pressure have been inconsistent. We examined the contributions of two AGT polymorphisms (T174M and M235T) to detect the effects of each on blood pressure variation, using single-site and two-site analyses. We analysed data from a study of 177 subjects from Accra, Ghana. We observed significant single-locus associations of the T174M polymorphism with average systolic (SBP) and diastolic blood pressure (DBP) when age was used as a covariate (P<0.001 and P=0.010, respectively). Also, we observed a significant association of the M235T polymorphism with SBPs and DBPs (P<0.001 and P=0.014, respectively). Finally, we observed a simultaneous significant association of the two polymorphisms with SBP and DBP (P<0.001 and P=0.026, respectively), although the two-loci model is not significantly better than either single-locus model. However, for SBP the two-loci model is marginally better (P=0.08 in comparison to both single-locus models). These results suggest that variants at these two AGT sites together, in conjunction with age, may be significantly associated with elevated SBP, whereas the single-site models are as good models of DBP. It is possible that earlier inconsistent results with these AGT polymorphisms with hypertension in African-derived populations may have resulted from an 'incomplete' model in the different study populations. Given the inconclusive nature of our two-loci results, this possibility requires further investigation. PMID- 15343354 TI - Dose-response of sodium excretion and blood pressure change among overweight, nonhypertensive adults in a 3-year dietary intervention study. AB - A cross-sectional dose-response relationship between sodium intake and blood pressure (BP) has been demonstrated, but evidence for a graded longitudinal effect is limited. Evaluation of BP response to sodium reduction was assessed in a 3-year lifestyle dietary intervention trial. BP changes at 18 and 36 months after enrollment were analysed according to concurrent quantitative changes in sodium excretion and by categories of success in sodium reduction among 1157 men and women, ages 30-54 years, with a diastolic BP (DBP) 83-89 mmHg, systolic BP (SBP) <140 mmHg, body weight 110-165% of sex-specific standard weight, and valid baseline urinary sodium excretion. Participants were randomized to a Sodium Reduction intervention (n=581) or Usual Care (n=576). From a 187 mmol/24 h baseline mean sodium excretion, net decreases were 44 mmol/24 h at 18 months and 38 mmol/24 h at 36 months in Sodium Reduction vs Usual Care. Corresponding net decreases in SBP/DBP were 2.0/1.4 mmHg at 18 months, and 1.7/0.9 mmHg at 36 months. Significant dose-response trends in BP change over quintiles of achieved sodium excretion were seen at both 18 (SBP and DBP) and 36 (SBP only) months; effects appeared stronger among those maintaining sodium reduction. Estimated SBP decreases per 100 mmol/24 h reduction in sodium excretion at 18 and 36 months were 2.2 and 1.3 mmHg before and 7.0 and 3.6 mmHg after correction for measurement error, respectively. DBP changes were smaller and nonsignificant at 36 months. In conclusion, incremental decreases in BP with lower sodium excretion were observed in these overweight nonhypertensive individuals. PMID- 15343355 TI - Associations of short-term weight changes and weight cycling with incidence of essential hypertension in the EPIC-Potsdam Study. AB - The aim of this study was to examine the relationships of short-term weight gain, weight loss, and weight cycling on the odds of developing hypertension. Normotensive middle-aged German men and women (n=12,362) of the European Prospective Investigation into Cancer and Nutrition-Potsdam Study were assigned to categories of 2-year short-term weight changes that were self-reported to have occurred prior to recruitment into the study (gain only, loss only, weight cycling, stable). After 2 years of follow-up after recruitment, 180 cases of incident essential hypertension were identified. In logistic regression models, odds ratios were estimated for the associations between short-term weight changes and risk of developing hypertension. Obesity status (BMI>or=30 or BMI<30 kg/m2) modified the associations between short-term weight change and incidence of diagnosed hypertension. Among obese individuals, short-term weight gain occurring during the 2 years prior to recruitment (OR=2.79, 95% CI 1.19-6.56), weight loss (OR=6.74, 95% CI 2.58-17.6) and weight cycling (OR=4.29, 95% CI 1.55-11.9) were strongly positively associated with incident hypertension, adjusted for age and gender, compared to obese individuals with short-term stable weight. No significant associations between short-term weight changes and risk of diagnosed hypertension were detected among non-obese individuals. Short-term weight changes appeared to present strong risk factors for developing hypertension among obese individuals. The effect seen for weight cycling supports the hypothesis that weight cycling increases the risk of hypertension. The finding for short-term weight loss may be explained by subsequent weight regain and needs further investigation. PMID- 15343356 TI - Blood pressure and atherosclerotic plaques in carotid, aortic and femoral arteries in elderly Finns with diabetes mellitus or impaired glucose tolerance. AB - The aim of the study was to evaluate the occurrence of atheromatous plaques in carotid, aortic and femoral arteries, focusing on blood pressure (BP). The study subjects consisted of 65-year-old Finns drawn from a population-based cohort. Ultrasonographic measurements were performed on 54 diabetic subjects, 97 subjects with impaired glucose tolerance (IGT) and 57 normoglycaemic subjects (NGT). High systolic BP (SBP) was defined as >or=160 mmHg and high diastolic BP as >or=95 mmHg. High pulse pressure (PP) was defined as the highest tertile (>or=75 mmHg) of PP and high mean BP (MBP) as the highest tertile (>or=111 mmHg) of MBP. The prevalence of atheromatous plaques was 77% (160/208) in carotid arteries, 94% (195/208) in aorta and 77% (161/208) in femoral arteries. A total of 64% (134/208) of the subjects had plaques in both carotid and femoral arteries, and they were compared with those who had plaques in 0-1 of these arteries. In addition to male gender and long-lasting smoking, the occurrence of plaques in both carotid and femoral arteries were associated with high SBP and high MBP. According to the results of multiple regression analyses, the adjusted odds ratio for plaques in both carotid and femoral arteries was 3.1 (95% CI 1.5-6.5) in subjects with high SBP compared to those with lower SBP. When SBP was replaced by high MBP, the adjusted odds ratio for it was 2.3 (95% CI 1.1-4.8). PMID- 15343357 TI - The 40th anniversary of the publication in 1964 of the first trial of the treatment of uncomplicated, severe hypertension by Hamilton, Thompson and Wisniewski. PMID- 15343358 TI - Sequential delivery of interferon-alpha gene and DCs to intracranial gliomas promotes an effective antitumor response. AB - Effective presentation of tumor antigens by dendritic cells (DCs) is considered to be essential for the induction of antitumor T-cell responses. Apoptotic and necrotic tumors have been noted to be a robust antigen source for DCs. Because glioma cells undergo apoptosis after transfection with the type I interferon (IFN) gene and type I IFNs promote the stimulatory activity of DCs, we hypothesized that transfection of glioma cells with type I IFN genes and provision of DCs would promote particularly effective antitumor activity by both facilitating apoptosis of glioma cells and the presentation of the glioma antigens, thereby inducing specific immune responses against glioma cells. We have previously reported the proof of this hypothesis in vitro and in a subcutaneous tumor model. Here we report an extension of this approach in intracranial (i.c.) gliomas using adenoviral IFN-alpha (Ad-IFN-alpha) vector. Mice bearing day-5 i.c. GL261 glioma received sequential intratumoral (i.t.) delivery of Ad-IFN-alpha and bone marrow-derived syngeneic DCs. This treatment prolonged survival in that nine of 17 animals survived long term (> 60 days versus 0 of 10 control animals). Specific CTL activity was demonstrated following this regimen in the cervical lymph nodes, and the therapeutic efficacy was dependent upon CD8+ cells. Furthermore, these animals were protected against subsequent re-challenge with GL261 gliomas. DCs injected i.t. survived in the tumor and migrated into cervical lymph node. In vitro migration assays revealed the ability of DCs to migrate toward the tumor, suggesting that i.t. injected DCs migrate through the glioma. Taken together, this combination of gene therapy and cellular immunotherapy may be an effective future strategy for treating human gliomas. PMID- 15343359 TI - Preclinical evaluation of a prostate-targeted gene-directed enzyme prodrug therapy delivered by ovine atadenovirus. AB - Gene-directed enzyme prodrug therapy (GDEPT) based on the Escherichia coli enzyme, purine nucleoside phosphorylase (PNP), provides a novel strategy for treating slowly growing tumors like prostate cancer (CaP). PNP converts systemically administered prodrug, fludarabine phosphate, to a toxic metabolite, 2-fluoroadenine, that kills PNP-expressing and nearby cells by inhibiting DNA, RNA and protein synthesis. Reporter gene expression directed by a hybrid prostate directed promoter and enhancer, PSMEPb, was assayed after plasmid transfection or viral transduction of prostate and non-CaP cell lines. Androgen-sensitive (AS) LNCaP-LN3 and androgen-independent (AI) PC3 human CaP xenografts in nude mice were injected intratumorally with an ovine atadenovirus vector, OAdV623, that carries the PNP gene under PSMEPb, formulated with cationic lipid for enhanced infectivity. Fludarabine phosphate was then given intraperitoneally for 5 days at 75 mg/m2/day. PNP expression was evaluated by enzymic conversion of its substrate using reverse phase HPLC. OAdV623 showed excellent in vitro transcriptional specificity for CaP cells. In vivo, expression of PNP persisted for > 6 days after OAdV623 injection and a single treatment provided 100% increase in tumor doubling time and > 50% inhibition of tumor growth for both LNCaP-LN3 and PC3 lines, with increased tumor necrosis and apoptosis and decreased tumor cell proliferation. OAdV623 significantly suppressed the growth of AS and AI human CaP xenografts in mice. PMID- 15343360 TI - Retroviral delivery of GAD-IgG fusion construct induces tolerance and modulates diabetes: a role for CD4+ regulatory T cells and TGF-beta? AB - Previous studies have demonstrated that antigen-specific tolerance could be induced by lipopolysaccharide (LPS)-stimulated B cells retrovirally transduced with an immunoglobulin-antigen (or epitope-containing peptide) fusion construct. To investigate the mechanism of this gene therapy system, we now adapted this approach to immunotherapy of spontaneous diabetes in nonobese diabetic (NOD) mice, a T-cell-mediated autoimmune disease triggered, in part, by a pathogenic response to glutamate decarboxylase (GAD) 65. We demonstrate that LPS-stimulated splenocytes, retrovirally transfected with GAD-IgG fusion construct, induce a significant antigen-specific hyporesponsiveness at both cellular and humoral levels and reduce the incidence of diabetes in female NOD mice. Parallel with disease protection, we observed a prolonged increase of the numbers of CD4+CD25+ T cells in the periphery of GAD-IgG-treated mice, compared to those treated with a control IgG vector, both in the prediabetic period and persisting even 8 months after gene therapy. This increase appeared to be induced by the repeated stimulation of the antigen in the periphery instead of a result of differentiation of T-cell precursor in the thymus. Moreover, CD4+CD25+ T cells induced by GAD-IgG fusion construct were capable of suppressing the proliferative response of CD4+CD25- T cells in vitro; and ablation of the activity of CD4+CD25+ T cells by blocking antibody against CD25 could reverse GAD-specific T-cell hyporesponsiveness. These results suggested that CD4+CD25+ T-cell subset induced in GAD-IgG-treated NOD mice represented the regulatory or suppressive CD4+CD25+ T cells (Treg) and might play an important role in the induction and maintenance of tolerance in NOD mice. Furthermore, the numbers of splenic CD4+CD62L+ regulatory T cells in GAD-IgG-treated mice during the prediabetic period and serum TGF-beta levels in 34-38-week-old GAD-IgG-protected mice were also increased, compared to control IgG-treated ones. Therefore, we propose that the induction of tolerance and the prevention of diabetes incidence in NOD female mice induced by the GAD IgG fusion construct may require CD4+ regulatory T cells, and the possible mediation of TGF-beta. PMID- 15343363 TI - TRUS and prostate biopsy: current status. PMID- 15343364 TI - Clusterin as a possible predictor for biochemical recurrence of prostate cancer following radical prostatectomy with intermediate Gleason scores: a preliminary report. AB - Disease recurrence following radical prostatectomy is a major concern in prostate cancer patients. Gleason scores are useful in predicting recurrence. Low Gleason scores are usually associated with long disease-free intervals, while high Gleason scores are suggestive of early recurrence. However, prediction of recurrence has been difficult with intermediate Gleason scores. Clusterin is a ubiquitous secretory sulfated glycoprotein. It is also an antiapoptotic mediator in prostate cancer. The objective of the present study is to determine if clusterin can serve as a predictive biomarker for recurrence of prostate cancer with intermediate Gleason scores in patients following radical prostatectomy. Prostatic specimens with Gleason score of 6 (3+3) or 7 (3+4) were obtained from the archival bank. Three groups of specimens were investigated. The first group was from nine patients who developed recurrent disease according to a persistent rise of serum PSA within 3 years following radical prostatectomy. Those in the second group and the third group were from patients who showed no evidence of disease recurrence for at least 5 y (11 patients) and 10 y (eight patients), respectively following the surgery. Histological sections were subjected to immunohistochemical staining using a monoclonal antibody specific for clusterin. The staining intensity was scored as 0, 1, 2, and 3, with 0 being no staining, 1 showing less than 25% positive staining, 2 being 25-50% positive, and 3 showing greater than 75% positive staining. One-way ANOVA with Bonferroni correction was used for statistical analysis. Evaluation of the scores of clusterin staining was carried out according to four specific areas in each specimen. They were (a) benign epithelial cells, (b) malignant epithelial cells (cancer epithelia), (c) stromal cells surrounding benign cells, and (d) stromal cells surrounding malignant cells (cancer stroma). Staining score in prostatic epithelial cells, benign as well as malignant, showed no significant relationship among the three patient groups. However, when staining scores in stromal cells were compared, there was a significant difference between patients with recurrent disease and those showed no evidence of disease recurrence for at least 10 y. Results of this preliminary study support the important role of clusterin in the stromal component for prostate cancer progression. Clusterin immunostaining may be useful to aid the prediction of chance of disease recurrence in patients with Gleason score 6 or 7 prostate cancer following radical prostatectomy. Further studies with a large number of cases are warranted to verify this preliminary finding. PMID- 15343366 TI - Tolerance, not immunity, crucially depends on IL-2. AB - Interleukin-2 (IL-2) was identified based on its potent T-cell growth-factor activity and is widely considered to be a key cytokine in T-cell-dependent immune responses. However, the main non-redundant activity of this cytokine centres on the regulation of T-cell tolerance, and recent studies indicate that a failure in the production of CD4(+)CD25(+) regulatory T cells is the underlying cause of autoimmunity in the absence of IL-2. In marked contrast to the importance of IL-2 in peripheral T-cell tolerance, T-cell immunity is readily elicited to various agents in the absence of IL-2 in vivo. Here, we discuss these findings and, in particular, the action of IL-2 on regulatory T cells and effector cells, and the targeting of IL-2 and/or the IL-2 receptor in clinical settings. PMID- 15343367 TI - TCR zeta-chain downregulation: curtailing an excessive inflammatory immune response. AB - The T-cell receptor (TCR) functions in both antigen recognition and signal transduction, which are crucial initial steps of antigen-specific immune responses. TCR integrity is vital for the induction of optimal and efficient immune responses, including the routine elimination of invading pathogens and the elimination of modified cells and molecules. Of the TCR subunits, the zeta-chain has a key role in receptor assembly, expression and signalling. Downregulation of TCR zeta-chain expression and impairment of T-cell function have been shown for T cells isolated from hosts with various chronic pathologies, including cancer, and autoimmune and infectious diseases. This review summarizes studies of the various pathologies that show this phenomenon and provides new insights into the mechanism responsible for downregulation of zeta-chain expression, its relevance and its clinical implications. PMID- 15343368 TI - Self-representation in the thymus: an extended view. AB - The thymus has been viewed as the main site of tolerance induction to self antigens that are specifically expressed by thymic cells and abundant blood-borne self-antigens, whereas tolerance to tissue-restricted self-antigens has been ascribed to extrathymic (peripheral) tolerance mechanisms. However, the phenomenon of promiscuous expression of tissue-restricted self-antigens by medullary thymic epithelial cells has led to a reassessment of the role of central T-cell tolerance in preventing organ-specific autoimmunity. Recent evidence indicates that both genetic and epigenetic mechanisms account for this unorthodox mode of gene expression. As we discuss here, these new insights have implications for our understanding of self-tolerance in humans, its breakdown in autoimmune diseases and the significance of this tolerance mode in vertebrate evolution. PMID- 15343369 TI - NALT- versus Peyer's-patch-mediated mucosal immunity. AB - Recent studies indicate that the mechanism of nasopharynx-associated lymphoid tissue (NALT) organogenesis is different from that of other lymphoid tissues. NALT has an important role in the induction of mucosal immune responses, including the generation of T helper 1 and T helper 2 cells, and IgA-committed B cells. Moreover, intranasal immunization can lead to the induction of antigen specific protective immunity in both the mucosal and systemic immune compartments. Therefore, a greater understanding of the differences between NALT and other organized lymphoid tissues, such as Peyer's patches, should facilitate the development of nasal vaccines. PMID- 15343370 TI - T-cell trafficking in asthma: lipid mediators grease the way. AB - Recruitment of T cells to the airways is crucial in the pathogenesis of asthma, and it is thought to be mediated mainly by peptide chemokines. By contrast, lipid mediators such as leukotrienes and prostaglandins have classically been thought to contribute to asthma pathogenesis by other mechanisms. However, as we discuss here, the recent molecular identification of leukotriene and prostaglandin receptors, as well as the generation of mice that are genetically deficient in them, has revealed that two of these lipids - leukotriene B(4) and prostaglandin D(2) - also direct T-cell migration and seem to cooperate with chemokines in a non-redundant, sequential manner to recruit T cells to the airways in asthma. PMID- 15343371 TI - Antiprion immunotherapy: to suppress or to stimulate? AB - Although human prion diseases are rare, they are invariably fatal, and treatments remain elusive. Hundreds of iatrogenic prion transmissions have occurred in the past two decades, and the bovine spongiform encephalopathy epidemic has raised concerns about prion transmission from cattle to humans. Research into therapeutics for prion disease is being pursued in several centres and prominently includes immunological strategies. Currently, the options that are being explored aim either to mobilize the innate and adaptive immune systems towards prion destruction or to suppress or dedifferentiate the lymphoreticular compartments that replicate prions. This article reviews the pathophysiology of prion diseases in mouse models and discusses their relevance to immunotherapeutic and immunoprophylactic antiprion strategies. PMID- 15343372 TI - Will telomere erosion lead to a loss of T-cell memory? AB - Evidence is accumulating that elderly individuals are more susceptible to infection with organisms to which they were previously immune. This indicates that there might be a limit to the persistence of immune memory. This fact is particularly disturbing because the average life expectancy of humans has almost doubled in the past 200 years and is still increasing. We discuss mechanisms that might constrain the persistence of memory T cells and consider whether humans will suffer from memory T-cell exhaustion as life expectancy increases. PMID- 15343374 TI - A new face in the line of defense against dementia. PMID- 15343375 TI - Dangerous liaisons at the virological synapse. AB - Cell-to-cell viral transmission facilitates the propagation of HIV-1 and human T cell leukemia virus type 1. Mechanisms of cell-to-cell transmission by retroviruses were not well understood until the recent description of virological synapses (VSs). VSs function as specialized sites of immune cell-to-cell contact that direct virus infection. Deciphering the molecular mechanisms of VS formation provides a fascinating insight into how pathogens subvert immune cell communication programs and achieve viral spread. PMID- 15343376 TI - Adenosine and ATP: traffic regulators in the kidney. AB - Glomerular filtration in the kidney is a continuous process that acts in concert with tubular reabsorption to prevent derangements of body fluid composition. Filtration is regulated by systemic factors, but it is also controlled by an intrinsic mechanism based on the anatomical connection between the distal nephron and the glomerular arterioles. Facing the threat of urinary salt loss, this mechanism causes vasoconstriction and reduces filtration by generating adenosine through the hydrolysis of nucleotide precursors such as 5'-AMP and possibly ATP. PMID- 15343377 TI - Why does diabetes increase atherosclerosis? I don't know! AB - There is a wealth of clinical data showing the relationship between diabetes mellitus and atherosclerosis and its clinical complications. To dissect this relationship, investigators have attempted, usually unsuccessfully, to create a small-animal model in which diabetes accelerates vascular lesion development. This effort has often been complicated by development of hyperlipidemia leading to difficulty in differentiating the effects of hyperglycemia from those of lipid abnormalities. A study in the current issue of the JCI provides data on a new mouse model in which atherosclerosis initiation is accelerated in diabetic mice and is reduced by insulin therapy. Moreover, these animals have greater intra arterial hemorrhage, which might be due to less stable plaques. PMID- 15343378 TI - Anti-C1q autoantibodies amplify pathogenic complement activation in systemic lupus erythematosus. AB - Patients with systemic lupus erythematosus (SLE) often develop glomerulonephritis (i.e., inflammation in the glomeruli of the kidney), commonly referred to as lupus nephritis. Patients with lupus nephritis typically have autoantibodies to the complement classical pathway protein C1q. Whether these anti-C1q antibodies play any role in the development of lupus nephritis has been unclear. In this issue of the JCI, a new study demonstrates that anti-C1q antibodies can amplify glomerular injury but only when they are bound within the glomerulus to C1q that has been already brought to that site by other types of glomerular-reactive autoantibodies. These studies are the first, to our knowledge, to provide a causal link between anti-C1q antibodies and target organ damage in SLE. PMID- 15343379 TI - Stat3 is required for the development of skin cancer. AB - Signal transducer and activator of transcription 3 (Stat3) is a transcription factor that is constitutively activated in a variety of human malignancies, including prostate, lung, brain, breast, and squamous cell carcinomas. Inhibition of activated Stat3 leads to decreased proliferation and apoptosis of many cancer derived cell lines, while the introduction of a constitutively activated form of Stat3 into immortalized human breast epithelial cells and rodent fibroblasts results in cellular transformation. Collectively, these data suggest a role for Stat3 in oncogenesis. A new study from Chan et al. is the first to demonstrate a requirement for Stat3 in de novo epithelial carcinogenesis in vivo. Using the two step model of chemically induced skin carcinogenesis, the authors demonstrated that mice deficient in Stat3 were completely resistant to skin tumor development. PMID- 15343380 TI - An amino-bisphosphonate targets MMP-9-expressing macrophages and angiogenesis to impair cervical carcinogenesis. AB - A mouse model involving the human papillomavirus type-16 oncogenes develops cervical cancers by lesional stages analogous to those in humans. In this study the angiogenic phenotype was characterized, revealing intense angiogenesis in high-grade cervical intraepithelial neoplasias (CIN-3) and carcinomas. MMP-9, a proangiogenic protease implicated in mobilization of VEGF, appeared in the stroma concomitant with the angiogenic switch, expressed by infiltrating macrophages, similar to what has been observed in humans. Preclinical trials sought to target MMP-9 and angiogenesis with a prototypical MMP inhibitor and with a bisphosphonate, zoledronic acid (ZA), revealing both to be antiangiogenic, producing effects comparable to a Mmp9 gene KO in impairing angiogenic switching, progression of premalignant lesions, and tumor growth. ZA therapy increased neoplastic epithelial and endothelial cell apoptosis without affecting hyperproliferation, indicating that ZA was not antimitotic. The analyses implicated cellular and molecular targets of ZA's actions: ZA suppressed MMP-9 expression by infiltrating macrophages and inhibited metalloprotease activity, reducing association of VEGF with its receptor on angiogenic endothelial cells. Given its track record in clinical use with limited toxicity, ZA holds promise as an "unconventional" MMP-9 inhibitor for antiangiogenic therapy of cervical cancer and potentially for additional cancers and other diseases where MMP-9 expression by infiltrating macrophages is evident. PMID- 15343381 TI - Impairment of tubuloglomerular feedback regulation of GFR in ecto-5' nucleotidase/CD73-deficient mice. AB - Adenosine coordinates organ metabolism and blood supply, and it modulates immune responses. In the kidney it mediates the vascular response elicited by changes in NaCl concentration in the macula densa region of the nephron, thereby serving as an important regulator of GFR. To determine whether adenosine formation depends on extracellular nucleotide hydrolysis, we studied NaCl-dependent GFR regulation (tubuloglomerular feedback) in mice with targeted deletion of ecto-5' nucleotidase/CD73 (e-5'NT/CD73), the enzyme responsible for adenosine formation from AMP. e-5'NT/CD73(-/-) mice were viable and showed no gross anatomical abnormalities. Blood pressure, blood and urine chemistry, and renal blood flow were not different between e-5'NT/CD73(+/+) and e-5'NT/CD73(-/-) mice. e 5'NT/CD73(-/-) mice had a significantly reduced fall in stop flow pressure and superficial nephron glomerular filtration rate in response to a saturating increase of tubular perfusion flow. Furthermore, whereas tubuloglomerular feedback responses did not change significantly during prolonged loop of Henle perfusion in e-5'NT/CD73(+/+) mice, a complete disappearance of the residual feedback response was noted in e-5'NT/CD73(-/-) mice over 10 minutes of perfusion. The contractile response of isolated afferent arterioles to adenosine was normal in e-5'NT/CD73(-/-) mice. We conclude that the generation of adenosine at the glomerular pole depends to a major extent on e-5'NT/CD73-mediated dephosphorylation of 5'-AMP, presumably generated from released ATP. PMID- 15343382 TI - Nef stimulates proliferation of glomerular podocytes through activation of Src dependent Stat3 and MAPK1,2 pathways. AB - In collapsing focal segmental glomerulosclerosis (FSGS) of HIV-associated nephropathy (HIVAN), podocytes exhibit a high proliferation rate and loss of differentiation markers. We have found previously that the nef gene of HIV-1 is responsible for these changes. Here, we investigated the signaling pathways induced by Nef and its role in the pathogenesis of HIVAN. Using conditionally immortalized podocytes after differentiation, we found that infection of podocytes with nef increased Src kinase activity and signal transducer and activator of transcription 3 (Stat3) phosphorylation and activated the Ras-c-Raf MAPK1,2 pathway. A dominant negative mutant of Src abolished the Nef effect, whereas inhibition of MAPK1,2 or dominant negative Stat3 reduced Nef effects partially. Reducing the expression of Nef with small interference RNA reversed the Nef effect. Mutation of Nef in the PxxP or R105R106 motifs diminished Nef signaling and the phenotypic changes in podocytes. Both phospho-MAPK1,2 and phospho-Stat3 staining increased in podocytes of kidneys from HIV-1 transgenic mice compared with their littermates and in podocytes of kidneys from HIVAN patients compared with HIV patients with non-HIVAN kidney diseases or non-HIV patients with idiopathic FSGS, classic FSGS, or minimal-change disease. These data suggest that Nef-induced activation of Stat3 and Ras-MAPK1,2 via Src dependent pathways is responsible for podocyte proliferation and dedifferentiation, a characteristic finding in collapsing FSGS of HIVAN. PMID- 15343383 TI - Hypothalamic PI3K and MAPK differentially mediate regional sympathetic activation to insulin. AB - The action of insulin in the central nervous system produces sympathetic nervous system activation (also called sympathoactivation), although the neuronal intracellular mechanisms that mediate this are unclear. We hypothesized that PI3K and MAPK, the major pathways involved in insulin receptor signaling, mediate sympathetic nerve responses to insulin. Intracerebroventricular administration of insulin in rat increased multifiber sympathetic nerve activity to the hindlimb, brown adipose tissue (BAT), adrenal gland, and kidney. Ex vivo biochemical studies of mediobasal hypothalamic tissue revealed that insulin stimulated the association of insulin receptor substrate-1 with the p85alpha subunit of PI3K and also tyrosine phosphorylation of p42 and p44 subunits of MAPK in the hypothalamus. In order to determine whether PI3K and/or MAPK were involved in insulin-mediated sympathoactivation, we tested the effect of specific inhibitors of PI3K (LY294002 and wortmannin) and MAPK (PD98059 and U0126) on regional sympathetic responses to insulin. Interestingly, regional sympathoactivation to insulin was differentially affected by blockade of PI3K and MAPK. Inhibition of PI3K specifically blocked insulin-induced sympathoactivation to the hindlimb, while inhibition of MAPK specifically blocked insulin-induced sympathoactivation to BAT. Sympathoactivation to corticotrophin-releasing factor, however, was not affected by inhibition of PI3K and MAPK. These data demonstrate that PI3K and MAPK are specific and regionally selective mediators of the action of insulin on the sympathetic nervous system. PMID- 15343384 TI - Diabetes and diabetes-associated lipid abnormalities have distinct effects on initiation and progression of atherosclerotic lesions. AB - Diabetes in humans accelerates cardiovascular disease caused by atherosclerosis. The relative contributions of hyperglycemia and dyslipidemia to atherosclerosis in patients with diabetes are not clear, largely because there is a lack of suitable animal models. We therefore have developed a transgenic mouse model that closely mimics atherosclerosis in humans with type 1 diabetes by breeding low density lipoprotein receptor-deficient mice with transgenic mice in which type 1 diabetes can be induced at will. These mice express a viral protein under control of the insulin promoter and, when infected by the virus, develop an autoimmune attack on the insulin-producing beta cells and subsequently develop type 1 diabetes. When these mice are fed a cholesterol-free diet, diabetes, in the absence of associated lipid abnormalities, causes both accelerated lesion initiation and increased arterial macrophage accumulation. When diabetic mice are fed cholesterol-rich diets, on the other hand, they develop severe hypertriglyceridemia and advanced lesions, characterized by extensive intralesional hemorrhage. This progression to advanced lesions is largely dependent on diabetes-induced dyslipidemia, because hyperlipidemic diabetic and nondiabetic mice with similar plasma cholesterol levels show a similar extent of atherosclerosis. Thus, diabetes and diabetes-associated lipid abnormalities have distinct effects on initiation and progression of atherosclerotic lesions. PMID- 15343385 TI - Maturational differences in lung NF-kappaB activation and their role in tolerance to hyperoxia. AB - Neonatal rodents are more tolerant to hyperoxia than adults. We determined whether maturational differences in lung NF-kappaB activation could account for the differences. After hyperoxic exposure (O2 > 95%), neonatal (<12 hours old) lung NF-kappaB binding was increased and reached a maximum between 8 and 16 hours, whereas in adults no changes were observed. Additionally, neonatal NF kappaB/luciferase transgenic mice (incorporating 2 NF-kappaB consensus sequences driving luciferase gene expression) demonstrated enhanced in vivo NF-kappaB activation after hyperoxia in real time. In the lungs of neonates, there was a propensity toward NF-kappaB activation as evidenced by increased lung I-kappaB kinase protein levels, I-kappaBalpha phosphorylation, beta-transducin repeat containing protein levels, and total I-kappaBalpha degradation. Increased lung p JNK immunoreactive protein was observed only in the adult lung. Inhibition of pI kappaBalpha by BAY 11-7085 resulted in decreased Bcl-2 protein levels in neonatal lung homogenates and decreased cell viability in lung primary cultures after hyperoxic exposure. Furthermore, neonatal p50-null mutant (p50(-/-)) mice showed increased lung DNA degradation and decreased survival in hyperoxia compared with WT mice. These data demonstrate that there are maturational differences in lung NF-kappaB activation and that enhanced NF-kappaB may serve to protect the neonatal lung from acute hyperoxic injury via inhibition of apoptosis. PMID- 15343386 TI - Anti-C1q autoantibodies deposit in glomeruli but are only pathogenic in combination with glomerular C1q-containing immune complexes. AB - Anti-C1q autoantibodies are present in sera of patients with several autoimmune diseases, including systemic lupus erythematosus (SLE). Strikingly, in SLE the presence of anti-C1q is associated with the occurrence of nephritis. We have generated mouse anti-mouse C1q mAb's and used murine models to investigate whether anti-C1q autoantibodies actually contribute to renal pathology in glomerular immune complex disease. Administration of anti-C1q mAb JL-1, which recognizes the collagen-like region of C1q, resulted in glomerular deposition of C1q and anti-C1q autoantibodies and mild granulocyte influx, but no overt renal damage. However, combination of JL-1 with a subnephritogenic dose of C1q-fixing anti-glomerular basement membrane (anti-GBM) antibodies enhanced renal damage characterized by persistently increased levels of infiltrating granulocytes, major histological changes, and increased albuminuria. This was not observed when a non-C1q-fixing anti-GBM preparation was used. Experiments with different knockout mice showed that renal damage was dependent not only on glomerular C1q and complement activation but also on Fcgamma receptors. In conclusion, anti-C1q autoantibodies deposit in glomeruli together with C1q but induce overt renal disease only in the context of glomerular immune complex disease. This provides an explanation why anti-C1q antibodies are especially pathogenic in patients with SLE. PMID- 15343387 TI - Cerebral vessel laminins and IFN-gamma define Trypanosoma brucei brucei penetration of the blood-brain barrier. AB - Subspecies of Trypanosoma brucei cause severe brain diseases after penetration of the blood-brain barrier. We investigated whether cytokines that modulate inflammatory cell infiltration into the brain also influence T. brucei neuroinvasion. Migration of a rodent pathogenic T. brucei strain from the cerebral blood vessels into the brain parenchyma was impeded in IFN-gamma(-/-), IFN-gamma receptor(-/-) (IFN-gammaR(-/-)), IL-12p40(-/-), and recombinant activating gene-1(-/-) (RAG-1(-/-)) mice as compared with their WT littermates despite higher levels of parasitemia in the mutant strains. Parasites accumulated in the perivascular compartment, confined between the endothelial and the parenchymal basement membranes, in certain areas of the brains of IFN-gamma(-/-), IFN-gammaR(-/-), and RAG-1(-/-) mice. This accumulation occurred around endothelial basement membranes containing the laminin alpha4 chain, while blood vessels showing robust laminin alpha5 chain immunostaining were not associated with parasite infiltration. The number of CD4+ and CD8+ T cells infiltrating the brain parenchyma was also reduced in the IFN-gamma(-/-) and IFN-gammaR(-/-) mice. Our findings suggest that lymphocyte-derived IFN-gamma facilitates trypanosome penetration across cerebral blood vessels and that the site of penetration is determined by the composition of the basement membranes of these vessels. PMID- 15343388 TI - Mouse retrovirus mediates porcine endogenous retrovirus transmission into human cells in long-term human-porcine chimeric mice. AB - Porcine endogenous retrovirus (PERV) is a potential pathogen in clinical xenotransplantation; transmission of PERV in vivo has been suggested in murine xenotransplantation models. We analyzed the transmission of PERV to human cells in vivo using a model in which immunodeficient NOD/SCID transgenic mice were transplanted with porcine and human lymphohematopoietic tissues. Our results demonstrate, we believe for the first time, that human and pig cells can coexist long-term (up to 25 weeks) without direct PERV infection of human cells. Despite the transplantation of porcine cells that did not produce human-tropic PERV, human cells from the chimeric mice were frequently found to contain PERV sequences. However, this transmission was due to the pseudotyping of PERV-C (a virus without human tropism) by xenotropic murine leukemia virus, rather than to de novo generation of human-tropic PERV. Thus, pseudotyping might account for the PERV transmission previously observed in mice. The absence of direct human cell infection following long-term in vivo coexistence with large numbers of porcine cells provides encouragement regarding the potential safety of using pigs that do not produce human-tropic PERV as source animals for transplantation to humans. PMID- 15343389 TI - The site of primary T cell activation is a determinant of the balance between intrahepatic tolerance and immunity. AB - Hepatic immunobiology is paradoxical: although the liver possesses unusual tolerogenic properties, it is also the site of effective immune responses against multiple pathogens and subject to immune-mediated pathology. The mechanisms underlying this dichotomy remain unclear. Following previous work demonstrating that the liver may act as a site of primary T cell activation, we demonstrate here that the balance between immunity and tolerance in this organ is established by competition for primary activation of CD8+ T cells between the liver and secondary lymphoid tissues, with the immune outcome determined by the initial site of activation. Using a transgenic mouse model in which antigen is expressed within both liver and lymph nodes, we show that while naive CD8+ T cells activated within the lymph nodes were capable of mediating hepatitis, cells undergoing primary activation within the liver exhibited defective cytotoxic function and shortened half-life and did not mediate hepatocellular injury. The implications of these novel findings may pertain not only to the normal maintenance of peripheral tolerance, but also to hepatic allograft tolerance and the immunopathogenesis of chronic viral hepatitis. PMID- 15343390 TI - EVI1 induces myelodysplastic syndrome in mice. AB - Myelodysplasia is a hematological disease in which genomic abnormalities accumulate in a hematopoietic stem cell leading to severe pancytopenia, multilineage differentiation impairment, and bone marrow (BM) apoptosis. Mortality in the disease results from pancytopenia or transformation to acute myeloid leukemia. There are frequent cytogenetic abnormalities, including deletions of chromosomes 5, 7, or both. Recurring chromosomal translocations in myelodysplasia are rare, but the most frequent are the t(3;3)(q21;q26) and the inv(3)(q21q26), which lead to the inappropriate activation of the EVI1 gene located at 3q26. To better understand the role of EVI1 in this disease, we have generated a murine model of EVI1-positive myelodysplasia by BM infection and transplantation. We find that EVI1 induces a fatal disease of several stages that is characterized by severe pancytopenia. The disease does not progress to acute myeloid leukemia. Comparison of in vitro and in vivo results suggests that EVI1 acts at two levels. The immediate effects of EVI1 are hyperproliferation of BM cells and downregulation of EpoR and c-Mpl, which are important for terminal erythroid differentiation and platelet formation. These defects are not fatal, and the mice survive for about 10 months with compensated hematopoiesis. Over this time, compensation fails, and the mice succumb to fatal peripheral cytopenia. PMID- 15343391 TI - Disruption of Stat3 reveals a critical role in both the initiation and the promotion stages of epithelial carcinogenesis. AB - Constitutive activation of signal transducer and activator of transcription 3 (Stat3) has been found in a wide spectrum of human malignancies. Here, we have assessed the effect of Stat3 deficiency on skin tumor development using the 2 stage chemical carcinogenesis model. The epidermis of Stat3-deficient mice showed a significantly reduced proliferative response following treatment with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) because of a defect in G1-to S-phase cell cycle progression. Treatment with the tumor initiator 7,12 dimethylbenz[a]anthracene (DMBA) resulted in a significant increase in the number of keratinocyte stem cells undergoing apoptosis in the bulge region of hair follicles of Stat3-deficient mice compared with nontransgenic littermates. Notably, Stat3-deficient mice were completely resistant to skin tumor development when DMBA was used as the initiator and TPA as the promoter. Abrogation of Stat3 function using a decoy oligonucleotide inhibited the growth of initiated keratinocytes possessing an activated Ha-ras gene, both in vitro and in vivo. In addition, injection of Stat3 decoy into skin tumors inhibited their growth. To our knowledge, these data provide the first evidence that Stat3 is required for de novo epithelial carcinogenesis, through maintaining the survival of DNA damaged stem cells and through mediating and maintaining the proliferation necessary for clonal expansion of initiated cells during tumor promotion. Collectively, these data suggest that, in addition to its emerging role as a target for cancer therapy, Stat3 may also be a target for cancer prevention strategies. PMID- 15343393 TI - Towards nuclear magnetic resonance micro-spectroscopy and micro-imaging. AB - The first successful experiments demonstrating Nuclear Magnetic Resonance (NMR) were a spin-off from the development of electromagnetic technology and its introduction into civilian life in the late forties. It was soon discovered that NMR spectra held chemically relevant information making it useful as an analytical tool. By introducing a new way of detection, moving away from continuous wave spectroscopy, Fourier Transform NMR helped to overcome sensitivity problems and subsequently opened the way for multi-dimensional spectroscopy. As a result NMR has developed into one of the most powerful analysis techniques with widespread applications. Still sensitivity is a limiting factor in the applicability of NMR. Therefore we witness a renaissance of technique development in magnetic resonance striving to improve its receptiveness. This tutorial review introduces the efforts currently made in miniaturizing inductive detection by designing optimal radio-frequency microcoils. A second approach is to introduce a new way of detecting magnetic resonance signals by means of very sensitive micromechanical force detectors. This shows that the detection limits in terms of absolute sensitivity or imaging resolution are still open to significant improvements. PMID- 15343392 TI - Increased DC trafficking to lymph nodes and contact hypersensitivity in junctional adhesion molecule-A-deficient mice. AB - Junctional adhesion molecule-A (JAM-A) is a transmembrane adhesive protein expressed at endothelial junctions and in leukocytes. In the present work, we found that DCs also express JAM-A. To evaluate the biological relevance of this observation, Jam-A(-/-) mice were generated and the functional behavior of DCs in vitro and in vivo was studied. In vitro, Jam-A(-/-) DCs showed a selective increase in random motility and in the capacity to transmigrate across lymphatic endothelial cells. In vivo, Jam-A(-/-) mice showed enhanced DC migration to lymph nodes, which was not observed in mice with endothelium-restricted deficiency of the protein. Furthermore, increased DC migration to lymph nodes was associated with enhanced contact hypersensitivity (CHS). Adoptive transfer experiments showed that JAM-A-deficient DCs elicited increased CHS in Jam-A(+/+) mice, further supporting the concept of a DC-specific effect. Thus, we identified here a novel, non-redundant role of JAM-A in controlling DC motility, trafficking to lymph nodes, and activation of specific immunity. PMID- 15343394 TI - Direct oxidation of haemoglobin at bare boron-doped diamond electrodes. AB - The communication reports the direct oxidation of human haemoglobin at a bare boron-doped diamond electrode under moderately alkaline conditions with detection limit of 0.4 microM. PMID- 15343395 TI - A procedure for calibration transfer between near-infrared instruments--a worked example using a transmittance single tablet assay for piroxicam in intact tablets. AB - A procedure was developed for different modes of calibration transfer in near infrared (NIR) spectroscopy, which included a method for the selection of a subset of samples appropriate for transfer. As a worked example, these guidelines were applied to the transfer of a multivariate calibration model, representing a validated NIR single tablet assay for the active within an intact pharmaceutical product, between three equivalent dispersive NIR transmission instruments. Transfer was first evaluated between two instruments, representing the situation where both were available during calibration development. A spectral correction method alone, applied to the transfer instrument, was not sufficient to facilitate transfer, with further optimisation of the calibration model using a novel wavelength selection algorithm necessary to remove regions of the spectral range that resulted in skewed predictions on the second instrument. Through this approach, a single calibration model was found to be equally accurate and precise on the two instruments. A procedure, using the Kennard-Stone algorithm, is described for determining a reduced number of samples as a transfer set using only the spectral information from the original instrument. The purpose of the subset was to permit transfer to a new instrument where that instrument was not available until after calibration development or where it was undesirable to re measure the full sample set (i.e. due to excessive reference chemistry). Utilising the transfer set, transfer to a third instrument was evaluated. The calibration model, optimised between the first two instruments, was not directly applicable for the third instrument, with further wavelength selection required to remove a small region of spectral data. On completion, using a full statistical evaluation, a single calibration model was found to be equally accurate and precise on all three instruments. PMID- 15343396 TI - MALDI-TOF MS detection of dilute, volume-limited peptide samples with physiological salt levels. AB - This paper presents a highly efficient sample preparation technique for matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The purpose of the research is to use a conventional MALDI support to directly and conveniently detect sub-nM levels of peptides from volume-limited samples with physiological salt levels. In this new method, highly uniform matrix nitrocellulose spots with a 500 microm diameter were conveniently generated by direct contact of a capillary tip to a stainless steel MALDI plate. An array of 50 microspots can be blotted from 1 microL matrix-nitrocellulose solution within 1 min. It was found that the addition of high concentration nitrocellulose to the alpha-cyano-4-hydroxycinnamic acid (CHCA) matrix solution is critical for the formation of microspots. Samples are deposited on top of those microspots and incubated for 3 min. The CHCA-nitrocellulose surface shows a significant peptide binding capability for sub-nM levels of peptide. Restricting the matrix spot diameter to 500 microm gives an analyte enrichment effect because the peptides are confined to a small solid-phase surface area. Selective peptide binding is seen even with >0.15 M salt levels. Loading small aliquots of samples with multiple applications allows low level peptide detection down to 100 pM. Push pull perfusates collected from the rat striatum were successfully analyzed with the microspot method. PMID- 15343397 TI - Development of a novel microfluidic immunoassay for the detection of Helicobacter pylori infection. AB - The miniaturization of laboratory processes offers substantial advantages over traditional techniques in terms of cost, speed, and potential for multistage automation. To date, only a few studies have reported successful microfluidics based immunoassays, most of which rely on fluorescence detection technologies. The goal of this study was to develop a poly(dimethylsiloxane) microfluidics based immunoassay methodology and a versatile colorimetric quantification scheme for the detection of visual colour changes resulting from immune reactions in microchannels. The novel immunoassay technique was applied towards the detection of Helicobacter pylori infection using 20 human serum samples of known infection status, and results compared with conventional nitrocellulose membrane-based dot ELISA. The microchannel immunoassay reliably detected H. pylori antigens in quantities on the order of 10 ng, which provides a sensitivity of detection comparable to conventional dot-blot assays. Sensitivity was 100%, specificity was 90%, positive predictive value 91%, and negative predictive value 100%, with an overall accuracy of 95%. The software developed generated results that were consistent with visual observations and by automatically taking into account background intensity changes, the software minimized subjectivity. Volumes of solutions used were 100-fold less compared with conventional immunoassays. Miniaturization of the ELISA using this technique provides a means for the accurate diagnosis of microbial infection while minimizing waste production. PMID- 15343398 TI - Elastomer-glass micropump employing active throttles. AB - We report a reciprocating microfluidic pump, the Micro Throttle Pump (MTP), constructed in a relatively uncomplicated manner from glass and microstructured poly(dimethylsiloxane)(PDMS). Unconventionally, the MTP employs throttling of fluid flow as distinct from fully-closing valve structures. Accordingly, this technique offers the prospect of solid-phase suspension tolerance. The reported MTP employs piezoelectrically (PZT) actuated deformation of flow constrictions (throttles) fabricated from PDMS at the two ports of a central, PZT actuated pump chamber. By appropriate time-sequencing of the individual PZTs' actuation, pumping can be induced in either direction. PDMS' elasticity further facilitates throttle operation by virtue of allowing significant PZT flexure that is substantially independent of the underlying PDMS microstructure. In contrast, in a rigid substrate such as silicon, deformation is constrained to where underlying microstructured cavities exist and this restricts design options. We describe the construction and performance of a prototype MTP capable of pumping 300 microl min(-1) or alternatively generating a back-pressure of 5.5 kPa. Preliminary modelling of MTP operation is also presented. PMID- 15343399 TI - Non-contact production of oligonucleotide microarrays using the highly integrated TopSpot nanoliter dispenser. AB - For the first time we report on the production of oligonucleotide microarrays using a highly parallel and highly integrated, pressure driven TopSpot nanoliter dispenser. The system enables non-contact printing of different media like oligonucleotides, DNA or protein solutions. We optimized the printing buffer needed for oligonucleotides microarrays production with respect to two major aspects: microfluidical optimum for droplet dispensing and biochemical coupling efficiency on different commercially available microarray slides. Coefficient of variations (CVs) of generated spot diameters were measured to be smaller than 1% within one single dispensing nozzle and smaller than 1.5% within all 24 parallel nozzles of the printhead for all printing buffers used. No carry-over and no cross-talk was found, in extensive experiments with oligonucleotides. Optimized printing buffer compositions and concentrations for oligonucleotide microarrays were found, as well as optimized coupling protocols. Furthermore, buffers and protocols were adapted to a host of different microarray slides used. With this system, prime critical points of microarray production are solved, leading to high quality high throughput microarray fabrication. PMID- 15343400 TI - Amino acid analysis using disposable copper nanoparticle plated electrodes. AB - A disposable copper nanoparticle-plated screen-printed carbon electrode (designated as Cun-SPE(100-nm)) provides a new material for the determination of native amino acids. All 20 underivatized amino acids can be sensitively determined at 0.0 V vs. Ag/AgCl in pH 8 phosphate buffer solution. The precisely controlled copper nanoparticles can boost up the CuIIO/CuI2O redox signal on the working surface without any prior pretreatment procedure. The formation of a reversible 1:1 CuIIO-amino acid complex on the Cun-SPE(100-nm) was proposed to play a key role in the reaction mechanism. Stable detection responses were obtained for all amino acids by flow injection analysis with detection limits (S/N = 3) that lie in the range of 24 nM-2.7 microM. Selected amino acids from six representative chemical natures were separated by HPLC and detected at the Cun-SPE(100-nm) with promising results. PMID- 15343401 TI - Identification of non-peptide species in selenized yeast by MALDI mass spectrometry using post-source decay and orthogonal Q-TOF detection. AB - The potential of tandem mass spectrometry following matrix-assisted laser desorption ionization (MALDI) was studied for speciation of selenium. Non-peptide selenium-containing compounds were isolated from a selenized yeast aqueous extract by size-exclusion chromatography. Post-source decay (PSD) was compared with orthogonal quadrupole collision cell dissociation for the purpose of obtaining fragmentation and structural information. In the PSD mode, the use of ion gate covering the whole isotopic cluster of the parent compound allowed the immediate recognition of fragments containing Se and those in which this element was absent. The tandem mass spectra obtained by orthogonal MALDI Q-TOF were equally informative in terms of the number of fragments but suffered from a poorer sensitivity. The mass accuracy was ca. 20 times better in the oMALDI configuration than in the PSD mode. An unknown selenium compound with an m/z 388 was detected with a mass accuracy of 3 ppm according to the proposed empiric formula. PMID- 15343402 TI - Ultra-smooth glass channels for bioassay with motor proteins. AB - Optically-flat glass channels were fabricated for the non-fluorescent observation of bio-molecules. Two bioassays of motor proteins were successfully performed, in which nanoscale beads and microtubules were clearly observed by a differential interference contrast (DIC) microscope and a dark-field microscope, respectively. The concentration of hydrofluoric acid (HF) was optimized to obtain an optically flat surface which was evaluated by AFM, SEM and dark-field microscopy. A glass channel was etched using a poly(dimethyl siloxane)(PDMS) microfluidic channel as a mask and sealed with a PDMS-coated coverslip. The channel volume of 2-3 microl realized the drastic reduction of proteins required per assay compared with a conventional flow cell method requiring 20 microl. PMID- 15343403 TI - Monitoring of recombinant survival motor neuron protein using fiber-optic surface plasmon resonance. AB - Spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality. SMA is caused by the homozygous loss of the survival motor neuron 1 (SMN1) gene. A nearly identical copy gene exists known as SMN2, however, due to an aberrant splicing event, the SMN2 gene fails to produce sufficient full-length protein to protect against disease development in the absence of SMN1. While a number of compounds have recently been identified that can stimulate full-length survival motor neuron (SMN) expression from the nearly identical copy SMN2, one of the difficulties has been the lack of a highly reproducible and quantitative means to measure the levels of SMN protein. To develop a technique that allows the rapid and highly sensitive measurement of SMN protein, a Surface Plasmon Resonance (SPR) application has been developed. The ability to quantify unassociated SMN protein and monitor the binding of SMN with other proteins in solution using a SPR sensor in less than 15 min and at low ng mL(-1) levels in HEPES Buffer Saline (HBS) has been achieved. The detection limit for the specific binding of SMN in HBS pH 7.4 solution is 0.99 ng mL(-1) with non-specific binding accounting for approximately 30% of the signal. Quantification of SMN is based on an immunoassay performed on the gold surface of the SPR sensor. 16-mercaptohexadecanoic acid (MHA) was reacted with dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) to form a pre-activated thiol (MHA-NHS). Antibodies for SMN were then coupled to the sensor with the pre-activated thiol. Sensor specificity was examined with mixtures of myoglobin (MG) and SMN. SMN sensor response decreases by more than 60% when MG was added to SMN. The decrease in sensor response can be attributed to non-specific binding of SMN to MG, verified with a sensor for MG. PMID- 15343404 TI - Spectrofluorimetric determination of surface-bound thiol groups and its application to the analysis of thiol-modified silicas. AB - A sensitive spectofluorimetric method has been developed for the quantitative measurement of surface-bound thiol groups. The procedure is based on the quantitative esterification of the thiol group with Rhodamine B and its subsequent release from the solid by base hydrolysis for spectrofluorimetric determination. Application of the method to the analysis of thiol-modified nanoparticulate silicas yielded results that compared favourably against alternative approaches based on measurements of mercury capacity and iodometric titration of the thiol groups. Non-specific Rhodamine uptake, assessed using unmodified silica and C18-modified silica, did not significantly influence the analytical results. When applied to a typical 50 mg sample, the detection limit of the procedure was 1 nmole SH g(-1) silica. PMID- 15343407 TI - Large diameter limbs for dilated common iliac arteries in endovascular aneurysm repair: is it safe? AB - In this prospective study we examined whether dilated common iliac arteries (CIAs) can provide a safe distal seal in endovascular aneurysm repair (EVAR) with the use of bifurcated stent grafts with large diameter limbs. Sixteen patients with 26 dilated CIAs with a diameter of > or =6 mm who were offered EVAR using stent grafts with large diameter limbs were included in the study (Group A). Forty-two patients who also underwent EVAR without iliac dilatation, matched for age, sex and surgical risk were used for comparison (controls-Group B). In group A mean CIA diameter was 18.2 mm (16-28) and mean abdominal aortic aneurysm (AAA) diameter was 6.87 +/- 1.05 cm; mean age was 77.2 +/- 4.8 yrs (67-81). Mean follow up was 33.6 months (2.8 yrs). CIA diameter changes and development of endoleaks were assessed by CT angiography (CTA). Overall iliac dilatation was present in 16/58 of our patients (27.6%). In 10 patients dilatation was bilateral (17.3%). Partial or complete flow to the internal iliac artery (IIA) territories was preserved in all patients post-EVAR. On follow-up, stable caliber of the dilated CIAs was observed in 21 patients (84%), enlargement of 1mm in 3 (16%), and failure of the distal attachment in 1 (6.2%). Compared to the control group there was no statistical significance in the incidence of complications. Dilated common iliac arteries provide a safe distal seal in patients who have undergone EVAR, thus obviating the need for additional endovascular procedures and sparing flow in the IIA vascular bed. PMID- 15343409 TI - Balint commentary questioned. PMID- 15343410 TI - Adherence among Mexican Americans with type 2 diabetes: behavioral attribution, social support, and poverty. PMID- 15343411 TI - Prevalence of insulin resistance syndrome in a primary health care center in Kuwait. PMID- 15343412 TI - Teaching learners to use mirroring: rapport lessons from neurolinguistic programming. PMID- 15343413 TI - Physicians' oral health education in Kentucky. AB - OBJECTIVES: The Physicians' Oral Health Education in Kentucky (POHEK) curriculum was developed to teach family medicine residents to (1) perform oral health screening and risk assessment and (2) recognize and manage common oral conditions for children ages 5 years and under. METHODS: Family medicine residents in urban and rural settings received didactics and hands-on experience providing oral screening, risk assessments, and counseling for their pediatric patients. EVALUATION: Residents were evaluated by comparing pretest and posttest means of surveys that assessed attitudes and knowledge. Chart audits were also performed. CONCLUSIONS: Residents' knowledge and attitudes improved in the oral health care of their pediatric patients. PMID- 15343414 TI - Thank you all for coming. PMID- 15343415 TI - "Doctor, are you listening?" A writing and reflection workshop. PMID- 15343416 TI - Entry of US medical school graduates into family medicine residencies: 2003-2004 and 3-year summary. AB - This is the 23rd report prepared by the American Academy of Family Physicians (AAFP) on the percentage of each US medical school's graduates entering family medicine residency programs. Approximately 9.3% of the 15,704 graduates of US medical schools between July 2002 and June 2003 were first-year family medicine residents in 2003, compared with 10.3% in 2002 and 10.9% in 2001. Medical school graduates from publicly funded medical schools were more likely to be first-year family medicine residents in October 2003 than were residents from privately funded schools, 10.6% compared with 7.0%. The West North Central and the Mountain regions reported the highest percentage of medical school graduates who were first-year residents in family medicine programs in October 2003 at 15.2% and 11.7%, respectively; the New England and Middle Atlantic regions reported the lowest percentages at 6.2% and 5.3%, respectively. Nearly half of the medical school graduates (46.1%) entering a family medicine residency program as first year residents in October 2003 entered a program in the same state where they graduated from medical school. The percentages for each medical school have varied substantially from year to year since the AAFP began reporting this information. This article reports the average percentage for each medical school for the last 3 years. Also reported are the number and percentage of graduates from colleges of osteopathic medicine who entered Accreditation Council for Graduate Medical Education-accredited family medicine residency programs, based on estimates provided by the American Association of Colleges of Osteopathic Medicine. PMID- 15343417 TI - Results of the 2004 national resident matching program: family medicine. AB - The results of the 2004 National Resident Matching Program (NRMP) reflect a leveling in the recent trend of declining student interest in family medicine residency training in the United States. Compared with the 2003 Match, 34 more positions (36 fewer US seniors) were filled in family medicine residency programs through the NRMP in 2004, at the same time as 14 fewer (four fewer US seniors) in primary care internal medicine, 10 more in pediatrics-primary care (one more US senior), and 35 more (38 more US seniors) in internal medicine-pediatric programs. In comparison, one less position (one more US senior) was filled in anesthesiology and seven fewer (five more US seniors) in diagnostic radiology, two "marker" disciplines that have shown increases over the past several years. Many different forces, including student perspectives of the demands, rewards, and prestige of the specialty; the turbulence and uncertainty of the health care environment; liability protection issues; and the impact of faculty and resident role models, continue to influence medical student career choices. A total of 165 more positions (12 more US seniors) were filled in categorical internal medicine while 164 more positions (15 more US seniors) were filled in categorical pediatrics programs, where trainees perceive options for either practicing as generalists or entering subspecialty fellowships, depending on the market. With the needs of the nation, especially for rural and underserved populations, continuing to offer opportunities for family physicians, family medicine experienced a slight increase through the 2004 NRMP. The 2004 NRMP suggests that the trend away from family medicine and primary care careers may be leveling off. PMID- 15343418 TI - American Academy of Family Physicians patient education materials: can patients read them? AB - BACKGROUND AND OBJECTIVES: A large proportion of US adults struggle with limited literacy skills, which affect their ability to fully function in health care settings. This study evaluated the readability of English language American Academy of Family Physicians (AAFP) patient education materials. METHODS: A random sample of 171 AAFP patient education materials was selected from a list of 518 health topics available via the Internet. The SMOG grade formula was used to measure readability of the written patient education materials. RESULTS: The mean SMOG grade level of AAFP patient education materials was 9.43 +/- 1.31 (range=6 12). This is higher than the average reading skills of US adults. CONCLUSIONS: Our results support and extend the findings of other readability studies across many medical specialties, all of which find that patient education materials are written at a difficulty level that is too high. The AAFP and individual family physicians should strive to improve communication with their patients by providing educational materials that are written at an appropriate reading level. PMID- 15343419 TI - Assessing health literacy in African American and Caucasian adults: disparities in rapid estimate of adult literacy in medicine (REALM) scores. AB - BACKGROUND AND OBJECTIVES: The influence of literacy on health and health care is an important area of investigation. Studies with a literacy focus are most valuable when literacy is assessed with psychometrically sound instruments. METHODS: This study used a prospective cohort sample of 1,610 primary care patients. Patients provided sociodemographics and took the Rapid Estimate of Adult Literacy in Medicine (REALM), a 66-item word pronunciation literacy test. RESULTS: The sample was 65% African American; 66% were men; 51% had a high school education or less. REALM scores were significantly related to education, age, and race but not gender. When stratified by education, differences between African Americans and Caucasians remained significant. Using 19 different strategies to shorten the 66-item instrument, reliability coefficients above.80 were maintained. CONCLUSIONS: The REALM is a robust assessment of health literacy. However, the discordance in scores between African Americans and Caucasians with similar educational attainment needs to be further addressed. A much shorter instrument would still have internally consistent scores and potentially be more useful in clinical settings. PMID- 15343420 TI - The effect of a literacy training program on family medicine residents. AB - BACKGROUND AND OBJECTIVES: Pediatric literacy promotion programs carried out in the primary care setting, such as Reach Out and Read (ROR), have been associated with improved language skills for preschool children. Primary care physicians have frequent contact with young families and may be well situated for a literacy promotion program for both children and adults. We examined whether introducing ROR and an adult literacy intervention improves family medicine residents' literacy knowledge, attitudes, and practices. METHODS: We conducted a single group pretest/posttest evaluation design study of residents in a family medicine residency program serving low-income families. Residents completed self administered questionnaires assessing literacy knowledge, attitudes, and practice. Then, through educational conferences, precepting, and ROR, residents were trained to assess and counsel patients about literacy. The same questionnaire was readministered 8 months later. RESULTS: All 24 (100%) residents completed both the pre- and post-intervention questionnaires. Literacy knowledge mean scores increased from 74.5% to 83.1%. After the intervention, residents reported a greater sense of comfort in counseling about childhood and adult literacy. After the intervention, a greater proportion of residents reported usually or always asking about literacy milestones (30.2% to 79.2%) and parent child reading (65.2% to 97.8%) during well-child visits. CONCLUSIONS: A family literacy promotion program improved family medicine residents' self-reported literacy knowledge, attitudes, and practices. Such interventions can be incorporated into the education of family medicine residents with meaningful results. PMID- 15343421 TI - Brief questions to identify patients with inadequate health literacy. AB - BACKGROUND AND OBJECTIVES: No practical method for identifying patients with low heath literacy exists. We sought to develop screening questions for identifying patients with inadequate or marginal health literacy. METHODS: Patients (n=332) at a VA preoperative clinic completed in-person interviews that included 16 health literacy screening questions on a 5-point Likert scale, followed by a validated health literacy measure, the Short Test of Functional Health Literacy in Adults (STOHFLA). Based on the STOFHLA, patients were classified as having either inadequate, marginal, or adequate health literacy. Each of the 16 screening questions was evaluated and compared to two comparison standards: (1) inadequate health literacy and (2) inadequate or marginal health literacy on the STOHFLA. RESULTS: Fifteen participants (4.5%) had inadequate health literacy and 25 (7.5%) had marginal health literacy on the STOHFLA. Three of the screening questions, "How often do you have someone help you read hospital materials?" "How confident are you filling out medical forms by yourself?" and "How often do you have problems learning about your medical condition because of difficulty understanding written information?" were effective in detecting inadequate health literacy (area under the receiver operating characteristic curve of 0.87, 0.80, and 0.76, respectively). These questions were weaker for identifying patients with marginal health literacy. CONCLUSIONS: Three questions were each effective screening tests for inadequate health literacy in this population. PMID- 15343422 TI - Health literacy: implications for family medicine. AB - As many as 90 million Americans have difficulty understanding and acting on health information. This health literacy epidemic is increasingly recognized as a problem that influences health care quality and cost. Yet many physicians do not recognize the problem or lack the skills and confidence to approach the subject with patients. In this issue of Family Medicine, several articles address health literacy in family medicine. Wallace and Lennon examined the readability of American Academy of Family Physicians patient education materials available via the Internet. They found that three of four handouts were written above the average reading level of American adults. Rosenthal and colleagues surveyed residents and found they lacked the confidence to screen and counsel adults about literacy. They used a Reach Out and Read program with accompanying resident education sessions to provide a practical and effective means for incorporating literacy assessment and counseling into primary care. Chew and colleagues presented an alternative to existing health literacy screening tests by asking three questions to detect inadequate health literacy. Likewise, Shea and colleagues reviewed the prospect of shortening the Rapid Estimate of Adult Literacy in Medicine (REALM), a commonly used health literacy screening tool. Both the Chew and Shea articles highlight the need for improved methods for recognizing literacy problems in the clinical setting. Further research is required to identify effective interventions that will strengthen the skills and coping strategies of both patients and providers and also prevent and limit poor reading and numeracy ability in the next generation. PMID- 15343426 TI - Prediction of impending rupture in aneurysms using 4D-CTA: histopathological verification of a real-time minimally invasive tool in unruptured aneurysms. AB - The authors describe the use of a 4D-CT angiogram to predict impending rupture in intact aneurysms, as a real-time, less invasive imaging technique. Histopathological verification and immunostaining of the bleb site performed on the study population reveals the significant predictive value of this tool. The point of maximum amplitude of pulsation of the aneurysm wall in unison with the RR interval of the electrocardiogram determines the potential rupture point. This helps in prioritizing the intervention for unruptured aneurysm cases, provides an effective screening of the high-risk population, and aids preoperative planning of clip application. PMID- 15343427 TI - Use of intraoperative dynamic infrared imaging with detection wavelength of 7-14 microm in the surgical obliteration of spinal arteriovenous fistula: case report and technical considerations. AB - BACKGROUND AND OBJECTIVE: Although improvements of spatial and temporal resolution in infrared (IR) imaging have enabled intraoperative real-time acquisition of physiological and pathological information on living organs, the imaging qualities of anatomical delineation of blood vessels and functional delineation of blood flow were insufficient to serve as visual monitoring. The main reason was partly due to the lack of an appropriate IR detection IR band (formerly 3 - 5 micro m), and the broad dynamic range in previous modalities. METHODS: To make a good contrast between blood vessels and surrounding tissues, the detection wavelength was shifted to the long-wave (7 - 14 micro m) part of IR spectrum, which includes the peak IR wave from living tissue (9 - 10 micro m), and the dynamic range was confined to +/- 10 degrees C around 35 degrees C. The novel camera system (IRIS IV infrared imaging system) was used for the visual monitoring of blood flow during the obliteration of a spinal perimedullary arteriovenous fistula at Th 7 in 71-year-old male patient. The temperature resolution of camera was 0.15 degrees C, with its intensity resolution of 16 bit (320 x 240 pixels), and data were stored at a rate of 30 frames/second. High quality delineation of blood vessels and blood flow was obtained all through the procedure without use of cold saline, which was an inevitable procedure to make contrast in the previously used thermography. However, the occasional use of an air blower was helpful to achieve adequate images in the deep and narrow area of the surgical field. The amount of shunt flow reduction was visualized by the intensity in the acquired imaging, which was also confirmed later by digital subtraction angiography. CONCLUSION: From the present experience, it is considered that this type of imaging may be able to substitute intraoperative blood vessel and blood flow monitoring in spinal and other neurosurgical disorders. PMID- 15343428 TI - Third ventriculostomy in communicating hydrocephalus in adult patients -- the role of lumbar and cranial cerebrospinal fluid outflow measurement. AB - OBJECTIVE: To evaluate the value of CSF outflow resistance testing as a criterion for endscopic treatment in case of communicating hydrocephalus. METHODS: 12 patients with communicating hydrocephalus were integrated in a prospective study from 1999 - 2001. Under the assumption of different hydromechanical properties of both spinal and cerebral CSF compartment the resorption capacity was measured. V. p. shunting, ventriculostomy or conservative treatment were performed dependent on the results and clinical outcome was evaluated. RESULTS: Six patients with pathological lumbar testing underwent primary ventriculo-peritoneal shunting. In 6 of the 12 cases the lumbar CSF resorption capacity was found to be normal and cerebral testing was accomplished. Two patients with normal spinal and cerebral resorption capacity were not operated. In the remaining 4 patients cerebral testing was found to be pathological and ventriculostomy was performed. An improvement of clinical symptoms was observed in patients operated with a v.-p. shunt and in the VCS group as well. CONCLUSIONS: This study demonstrates that there is a subgroup of patients with morphological communicating hydrocephalus that is characterized by a functional dissociation of hydromechanical properties of intracranial and spinal CSF compartment. In cases of regular CSF resorption but restricted CSF outflow from the ventricular system to the subarachnoid space ventriculo-peritoneal shunting may be an efficient therapy, but VCS was proved to be an efficient alternative. PMID- 15343429 TI - Hemorrhagic vascular complications of endoscopic transsphenoidal surgery. AB - Two hundred and fifty consecutive patients operated on by an endoscopic endonasal transsphenoidal approach were retrospectively analyzed in order to evaluate hemorrhagic vascular complications occurring during or after the surgical procedure and their appropriate management. Vascular complications of endoscopic transsphenoidal surgery are identical to those of a microsurgical transsphenoidal approach. Damage to the sphenopalatine artery and to the internal carotid artery (ICA), which are the most frequent vascular troubles, may require technical tricks because of some aspects connected to the approach itself and of the physical properties of the endoscope. Furthermore, the progress in interventional neuroradiology in the last decades offers new solutions in respect to the past, where the use of the surgical microscope was already a tremendous progress. The anatomic substrate of each complication is discussed, along with the peculiar surgical details related to it. PMID- 15343430 TI - Endoscopic third ventriculostomy: anesthetic implications. AB - Endoscopic third ventriculostomy (ETV) is one of the recent neurosurgical advances for the treatment of obstructive hydrocephalus. There has been number of publications, which have established the role of ETV in neurosurgical practice, particularly in hydrocephalus. ETV has developed into a therapeutic alternative to shunting for the management of patients with non-communicating hydrocephalus. This procedure requires a general anesthetic and necessitates violation of the brain parenchyma and manipulation via neural structures to access the floor of the third ventricle. This discussion will focus on the anesthetic implications during ETV. PMID- 15343431 TI - Waterjet dissection of gliomas -- experience with 51 procedures. PMID- 15343432 TI - Functional and diffusion-weighted magnetic resonance imaging for visualization of the postthalamic visual fiber tracts and the visual cortex. AB - Diffusion-weighted magnetic resonance imaging (MRI) offers the possibility to study the course of the cerebral white matter tracts whereas functional MRI (fMRI) provides information about the specific functions of cortical areas. We evaluated the combination of fMRI and diffusion-weighted MRI to detect cortical visual areas with their corresponding visual fiber tracts in 15 healthy controls (age: 23 - 53 years, male : female = 8 : 7). We demonstrated activation within the primary visual cortex and white matter bundles connecting the lateral geniculate body and the striate cortex in all subjects investigated. Additional activation could be appreciated in some subjects within the lateral geniculate bodies (n = 2) and the motion-sensitive area V5 (n = 3). The combination of diffusion-weighted and functional imaging allows visualization of the origin, direction and functionality of large white matter tracts. This will prove helpful for imaging structural connectivity within the brain during functional imaging. Moreover, this technique might provide important information for neurosurgical patients presenting with space-occupying lesions close to the cortical and subcortical visual system since this technique can -- in contrast to diffusion tensor imaging -- easily be adopted into a neuronavigation system and can be performed on all MR scanners capable of diffusion-weighted imaging without specific post-processing programs. PMID- 15343433 TI - Successful staged treatment for ruptured blister-like dissecting aneurysm of the intracranial internal carotid artery: acute GDC embolization for the blister-like aneurysm followed by proximal occlusion with extracranial-intracranial bypass in the chronic stage. AB - The surgical treatment of ruptured blister-like dissecting aneurysm on the internal carotid artery (ICA) is still controversial. We report a case of this disease successfully managed by a staged treatment: GDC packing into the blister like aneurysm in the acute stage followed by proximal occlusion in the chronic stage. The merit of this staged treatment is to prevent rerupture in the acute stage and to allow the proximal occlusion in the chronic stage with or without an extracranial-intracranial bypass, after assessment of tolerance of the ICA occlusion. PMID- 15343434 TI - Presurgical ultrasound-assisted neuro-examination in the surgical repair of peripheral nerve injury. AB - In spite of electrodiagnostic examinations, the determination of the precise localization of the injured site along the involved peripheral nerve may remain obscure or uncertain. Before starting the operation, a surgeon should have knowledge about the type of injury, the position of the proximal and distal nerve stumps, and the presence or absence of a neuroma and excessive perilesional scar tissue formation for orientation and planning of the surgical intervention. We hypothesized that real-time ultrasound could be helpful in the determination of the type of injury, the localisation of proximal and distal nerve stumps, as well as for diagnosing a neuroma. Fourteen patients with traumatic peripheral nerve injuries that were verified by neurological examinations and electrodiagnostic tests underwent surgical repair, and were examined by ultrasound before and during the surgical intervention. Visualisation of the injured site, the type of the injury, the position of the nerve stumps and the diagnosis of the neuroma were reliably feasible in all the patients by using ultrasonography. Axonal swelling of a nerve was diagnosed in 4 (29 %) patients, a stump neuroma was diagnosed in 3 (21 %) patients, a total nerve interruption (neurotmesis in the Seddon classification) was diagnosed in 9 (64 %) patients, and surrounding scar tissue was diagnosed in 5 (35 %) patients. Presurgical and intraoperative ultrasound-assisted neuroexamination is a useful diagnostic method in the determination of the precise localisation of the injured site, the type of injury, the position of stumps, and the diagnosis of a neuroma. The use of preoperative and intraoperative ultrasound can enhance the orientation of the surgeon to the surgical field. The application of our method to our patients shows that presurgical ultrasonographic neuroexamination can be used in the surgical repair of peripheral nerve injury. PMID- 15343435 TI - Endonasal endoscopic repair of cerebrospinal fluid rhinorrhea. AB - Cerebrospinal fluid (CSF) rhinorrhea implies a communication between the subarachnoid space and the upper respiratory tract. Trauma and postoperative defects are the more common causes of CSF rhinorrhea. The authors review their results with endoscopic repair of skull base defects associated with CSF rhinorrhea involving the paranasal sinuses. A total of 10 patients, 7 males and 3 females, were treated under endoscopic vision from 1997 to 2001. The causes of CSF rhinorrhea were traumatic (7), postoperative (2) and spontaneous (1). Four patients had the diagnosis and the site confirmed after intrathecal fluorescein saline injection. The obliteration of the CSF leak was achieved with fat free, mucoperichondrial, or mucoperiostal free grafts taken from the middle or inferior turbinate and kept in place by fibrin glue. Primary closure was obtained in 10 patients and one patient developed a recurrence 14 months later. The repair of the CSF rhinorrhea by endonasal endoscopic surgery is safe, very effective and is a valid alternative to the cranial approach. PMID- 15343436 TI - Endoscopic-assisted sciatic nerve exploration. AB - A sciatic nerve lesion is uncommon. We report the utility of the endoscope in an 18-year-old female with multiple lesions of her sciatic nerve. Somatosensory evoked potentials and MRI were indicative of a sciatic nerve lesion. We describe a simple technique of endoscopic exploration of the sciatic nerve to assist intraoperative decision making. PMID- 15343437 TI - Minimally invasive approaches to treat simultaneous occurrence of glioblastoma multiforme and intracranial aneurysm -- case report. AB - Primary brain tumors associated with intracranial aneurysm are rare. A combination of glioblastoma multiforme (GBM) with cerebral aneurysm is even more rare. We present a 67-year-old female, who suffered from progressive weakness of the right limbs and dysphasia for 2 days. She was referred from another hospital with impression of having a brain tumor. The non-enhanced computed tomographic (CT) scan of the brain showed a mass lesion over the left parietal region, and another tiny lesion with relatively high density over the left suprasellar region. After admission, the carotid angiogram revealed a 13 x 12 x 14 mm aneurysm with a base about 8 mm from the anterior wall of the left supraclinoid internal carotid artery (ICA) between the left ophthalmic artery and the left anterior choroid artery. We first clipped the aneurysm by a basal pterion keyhole approach. One week later, the tumor was en bloc removed by a small left parietal craniotomy under the guidance of a navigator. The final diagnosis was confirmed by histopathological examination as GBM. After the surgeries, the patient received palliative radiotherapy with a dose of 4000 cGy. She recovered well with no evidence of recurrence of the tumor one year after the operations. In this report, the careful diagnosis and the possible mechanisms of concurring GBM and cerebral aneurysm, as well as the strategies of surgical treatment involving minimally invasive approaches are discussed. PMID- 15343438 TI - Anterior cervical foraminotomy for unilateral spondylotic radiculopathy. AB - Anterior cervical foraminotomy (ACF) was developed under the concept of functional spine surgery, which directly eliminates compressive pathological factors while preserving functional anatomic features. The authors reviewed their results to determine the efficacy of the approach for unilateral cervical spondylotic radiculopathy (CSR). Nineteen patients were treated with the ACF with a follow-up from 12 to 36 months. There were 10 men and 9 women (mean age 49.8). Fourteen patients had a single ACF, and 5 had procedures at adjacent levels. The procedure involves microsurgical removal of the lateral portion of the uncinate process to identify the nerve root. Seventeen patients (89.5 %) were symptom-free or clearly improved, one (5.3 %) was unchanged and one patient (5.3 %) was worse in the visual analogue scale (VAS) score for radicular pain. One patient had developed contralateral foraminal stenosis at the level of the surgery and had undergone anterior discectomy and fusion. ACF provided good or excellent outcomes, with minimal morbidities, for patients with CSR. The advantages of ACF include direct decompression of the nerve root, and the preservation of the intervertebral disc and the motion segment. Thus, fusion-related sequelae, including graft-related complications, graft site complications and the adjacent level disease are avoided. The ACF procedure appears to be a good alternative for carefully selected patients with unilateral CSR. PMID- 15343439 TI - Novel retractor for endoscopic and microsurgical spinal interventions. AB - Retractors play an important role in surgery, especially using endoscopic techniques like microendoscopic discectomy. As a consequence of the minimally invasive approach the working space is frequently very restricted. All currently available systems still use tubular retractors with limitations regarding flexibility, stability and accessible working space. This paper discusses the technical aspects of a newly designed circular retractor system for endoscopic and microsurgical spinal interventions (SpineGate) and presents first clinical results in a series of 20 consecutive patients. The autoclavable system consists of a circular base plate and several valves varying in length. The variable fixation of the valves from different directions within the 360 degrees geometry of the base plate allows a continuously adjustable working space, depending on the individual anatomy. An additional fixation device is not necessary. The retractor is suitable for microendoscopic and microsurgical techniques. It is easy and safe to handle. The retractor system was successfully used in all cases without complications or instrumental malfunctions. All patients experienced good to excellent relief of their preoperative symptoms. PMID- 15343440 TI - Intraoperative bradycardia and postoperative hyperkalemia in patients undergoing endoscopic third ventriculostomy. PMID- 15343441 TI - [Ocular traumatology]. PMID- 15343442 TI - [Lacrimal lacerations--what is to be done?]. PMID- 15343443 TI - [Lacrimal system injuries -- primary and secondary surgical care]. AB - BACKGROUND: Canalicular lacerations can be the result of sharp or blunt trauma as well as burns of the facial region. The nasolacrimal duct may become obstructed as an after-effect of naso-orbital trauma. MATERIAL AND METHODS: Main principles of surgical repair of the lacrimal system are reviewed with regard to the outcome of our own patients compared with the literature. RESULTS: Success rates in the primary repair of traumatic injuries of canalicular system are about 70 to 82 %. 14 of 20 patients who were managed between 1976 and 1980 in our department were cured permanently. 55 of 272 secondary reconstructions of canalicular obstructions (1976 - 1997) were caused by trauma. Conjunctivodacryocystorhinostomy with polyethylene or silicone tubes (1978 - 1999, n = 37, 21 traumatic) had success rates of 61 % and 72 %, respectively. 11 (4.8 %) of 228 dacryocystorhinostomies (1991 - 2000) were necessary as a result of traumatic injuries and achieved a success rate of 87.5 %. CONCLUSIONS: Canalicular lacerations need urgent primary repair with silicone intubation and special heed to the medial canthal tendon. The success rate of secondary reconstructions of traumatically caused tear-duct system obstructions depends on the microsurgical techniques that were used. PMID- 15343444 TI - [Traumatology of the eyelids--what is to be done for the initial care?]. AB - We describe the injuries of the eyelids depending on the reason, type and severity of the trauma. We are giving a diagnostical guide. By references to the anatomical specialties of the eyelids we deliver therapeutical recommendations for the different types of injuries. PMID- 15343445 TI - A proactive treatment approach for eyes with perforating injury. AB - BACKGROUND: Perforating injuries continue to have a poor prognosis with less than two-thirds of eyes having at least ambulatory final vision, due to proliferation originating from the exit wound. MATERIALS AND METHODS: We developed a new, proactive treatment method, which is applicable in most eyes with perforating trauma. The strategy involves limited, indirect ophthalmoscopic vitrectomy during the primary repair; heavy topical corticosteroid therapy postoperatively; and complete vitrectomy on day 3, including prophylactic retinectomy around the exit wound, evacuation of subretinal blood, laser retinopexy, and silicone oil implantation. RESULTS: In the five consecutive eyes in which the proactive treatment approach was used, no "enscarceration" of the retina into the exit wound, proliferative vitreoretinopathy, retinal detachment, or retinal folding has occurred. The median visual acuity improved from count fingers to 0.6 in the three eyes without macular involvement. CONCLUSIONS: Far-reaching conclusions must not be made based on such a small series, but the proactive treatment approach appears promising in preventing proliferation-related complications such as tractional retinal detachment or retinal fold development. A similar approach should be considered for eyes with deep impact trauma from intraocular foreign bodies and in eyes with retinal incarcerated retina in the rupture wound. PMID- 15343446 TI - [Epidemiology of open globe eye injuries: analysis of 1026 cases in 18 years]. AB - BACKGROUND: Ocular trauma has significant impact on the patient's future quality of life. Progress in the treatment of ocular trauma may ameliorate the outcome, but preventive measures are more effective. However, prevention of ocular trauma can only be effective when the current epidemiology of ocular trauma is known. Therefore, we have now collected data on ocular trauma over nearly 20 years. This paper provides an overview on the development of open globe injuries in the past 20 years. PATIENTS: The records of 1026 patients with open globe injuries who were primarily treated at the Universities of Freiburg and Wurzburg between January 1, 1981 and December 31, 1999, were sufficiently analyzed. The following parameters were evaluated: age, extent of injury, sex, cause. and activity at the time of injury. RESULTS: After correction for demographic distribution, the risk for open globe injury was highest for young adults and lowest for seniors. In recent years, the risk for severe eye injury is more equally distributed and is increasing for old people. We noted decreasing injuries at work and during traffic accidents, especially for young female front-seat passengers. On the other hand, we observed an increasing proportion of domestic eye injuries related to hobby activities. With increasing age we noted more posterior segment injuries. 9 % of all open globe injuries occurred among seniors (> 65 years old), but 41 % of all ruptures occurred in this age group. Preceding cataract surgery was identified as a risk factor. 38 % of injured persons in this age group had already had intraocular surgery. After the introduction of small incision techniques at the beginning of the 1990 s, the frequency of globe ruptures began to decrease again. 3.6 % of the injured eyes developed an endophthalmitis, in agricultural injuries the rate of endophthalmitis was 12 %. CONCLUSION: There is a relative constant incidence of 3.0 open globe injuries per 100,000 population. The circumstances of the injuries underwent major changes within the period of these observations. PMID- 15343447 TI - [Eye trauma in childhood and youth]. AB - Childhood trauma of the eye and its adnexa represents approx. 4 - 20 % of all eye injuries. Due to ambiguous patient history and limited cooperation a comprehensive diagnosis of pediatric eye trauma is often difficult. The weakness of the infantile cornea, the thin and elastic anterior lens capsule, and the firm vitreous with its strong adherence to the retina bear particular intra- or postoperative problems. Moreover, the quick development of secondary cataract and the high risk of PVR may complicate the posttraumatic and postoperative course. Small infants represent a special subgroup in various regards mainly because of the risk of amblyopia which often causes functional loss in spite of a good anatomical reconstruction. The relative frequency of certain injuries differs between children and adults. The ocular manifestation of child abuse as well as bite and fireworks injuries are characteristic for childhood and will be discussed more in detail. PMID- 15343448 TI - [Surgical management of secondary angle closure glaucoma after trauma]. AB - Medical and surgical treatment of secondary angle closure glaucoma has often been disappointing. Therefore the visual prognosis of these eyes is mostly restricted. Transscleral cyclophotocoagulation is a relatively safe method for the treatment of advanced refractory glaucoma and represents the method of choice in secondary angle closure glaucoma due to anterior peripheral synechiae. In younger glaucoma patients and patients with traumatic glaucoma, long-term reduction of the intraocular pressure is only partially achieved. New surgical techniques will increase the possibilities of an effective reduction of the intraocular pressure in secondary angle closure glaucoma. On the other hand, data concerning the clinical efficacy and safety are still limited. These new procedures are endoscopic cyclophotocoagulation, retinectomy and the implantation of drainage devices via the pars plana. Further evaluation and modifications of these surgical techniques should markedly improve the visual prognosis of eyes with secondary angle closure glaucoma. PMID- 15343449 TI - [Contact lens fitting as a possibility for visual rehabilitation in patients after open globe injuries]. AB - INTRODUCTION: Visual rehabilitation after open globe injury may be a challenging process because of ametropia following aphakia, corneal scarring with high or irregular corneal astigmatism or loss of contrast sensitivity due to traumatic aniridia. We report on contact lens fitting for visual rehabilitation in patients after open globe injury. PATIENTS: From 2000 to 2003, contact lenses were fitted unilaterally for the visual rehabilitation in 13 patients after open globe injury. In three patients we found unilateral aphakia, in 8 patients a high or irregular astigmatism after penetrating or autorotation keratoplasty and in two patients a traumatic aniridia, in one case combined with aphakia. RESULTS: 11 rigid contact lenses were fitted with different designs of the front and back surface as well as two iris-print lenses. In 11 patients (86 %) a good visual rehabilitation was achieved with an increase of visual acuity up to 9 lines while obtaining a good contact lens tolerance. One patient wearing an iris-print contact lens was unable to tolerate the contact lens due to its thickness and its weight. In another patient fitting of a contact lens was not possible because of the complicated corneal condition. We did not observed severe contact lens complications at any time. CONCLUSIONS: In addition to operative procedures for visual rehabilitation after open globe injuries, the use of contact lenses is another possible procedure for refractive correction. Different problems such as ametropia following aphakia, irregular or high astigmatism or aniridia can be solved with good visual results, good tolerance and less complications. PMID- 15343450 TI - [Prognosis of corneal transplantation after penetrating eye injury]. AB - BACKGROUND: The success of penetrating keratoplasty (PKP) after penetrating eye injuries is often worse than in keratoplasty with Fuchs' endothelial dystrophy. However, comparable investigations are rare. Therefore, we have retrospectively analyzed the results and complication rates of PKPs in patients who experienced penetrating eye injuries. PATIENTS AND METHODS: Between 1990 and 1997 forty-four patients who received a corneal transplant at Kiel University Eye Hospital after penetrating eye injury (study group S) were analyzed. 29 patients were re examined (visual acuity testing, slit lamp microscopy, applanation tonometry, stereo testing, corneal TMS topography, keratometry, endothelial microscopy, perimetry, indirect retinoscopy, testing of the retinal visual acuity and probatory contact lens fitting). Postoperative follow-up ranged between 1 and 6 years (median 3 years). The results were compared to a group of corneal grafts performed in patients with Fuchs' endothelial dystrophy (K, n = 18 patients, 25 eyes, median follow-up 2.8 years). RESULTS: 38males and 6 women had had a bilateral visual acuity of 0.8 or better before the accident, except for three cases. At the time of eye injury the patients' age ranged from 4 to 76 years. The eye trauma was caused by splinters of metal (45 %), glass (20 %), wood (16 %) or stone (11 %). In 73 % of the cases the injury was confined to the anterior segment of the eye (S1). The posterior part of the eye was involved in 27 % (S2). BCVA of S before keratoplasty (KP) was 1/15 on average (K 0.2), at least one year after KP 0.25 (K 0.4); post KP S1 was better than S2 (p = 0.0234), and K better than S (p = 0.0009). In group S 29 patients received a graft at random because of a central corneal scar, 14 patients received an HLA-typed graft with 2.3 mismatches on average. 93 % of the grafts (41 patients) were clear at the last examination, 8 patients showed rejection episodes, half of them were reversible. 5 patients received a second graft because of an irreversible rejection or endothelial decompensation. BCVA improved by at least one line in 84 % in S (K 92 %). BCVA was unchanged after KP in 6.8 % (K 8 %). In 9.1 % of S BCVA after KP was worse (K 0 %). 32 out of 44 patients had a BCVA of 0.1 or better after the last check-up, 13 out of 44 patients presented with a BCVA of 0.5 or better (K40 %). 15 out of 29 patients separately tested showed stereopsis compared to 8 preoperatively in S. In 48 % of the patients studied stereovision could not be restored. 82 % of the patients suffered from topographical irregular astigmatism in S, the superficial geometry of recipient's cornea seems relevant for astigmatism of the graft. Frequent postsurgical complications consisted of secondary glaucoma (S 27 %, K 4 %), immunological rejections (S 18 %, K 0 %), and amblyopia (S 14 %, K 0 %). CONCLUSION: Compared to patients with Fuchs' dystrophy the results of grafting after eye injuries are significantly worse. This relates to BCVA, astigmatism, frequency of secondary glaucoma and graft rejection. PMID- 15343451 TI - [Anti-infectives in eye injuries]. AB - BACKGROUND: We present the current concepts for the use of anti-infectives parallel to surgery of traumatic eye injuries. MATERIAL AND METHODS: Data from the national and international literature were collected using the PubMed, Medline, Drugs and Pharmacology, ISI, and Cochrane databanks. In addition, the experience from our tertiary center was taken into consideration. RESULTS: For palpepral and globe injuries prophylactic and therapeutic anti-infectives are recommended as supporting measures to surgical repair. These include antiseptics (PVP-iodine), antibiotics (cefuroxime, vancomycin, ceftazidime), antimycotics (voriconazole) and anti-inflammatory substances (prednisolone). CONCLUSIONS: Several drugs are useful for the prevention and/or treatment of infection following eye injury. PMID- 15343452 TI - [Vaccination for stab and bite injuries in the facial region]. AB - In cases of stab and bite injuries to the face there is a risk of general infection and the necessity for an active and/or passive immunization must be considered by the first treating physician. In Central Europe tetanus, rabies, hepatitis B and C as well as HIV must be taken into account as possible consequences of such injuries. With regard to a tetanus immunization the indication for the post-exposition prophylaxis (PEP) should generally be as wide as possible. For other protective vaccinations a differentiated decision in co operation with other fields of activity and public health authorities should be realized. The current recommendations for the systemic infections relevant in Central Europe are described. PMID- 15343453 TI - [Can the optic nerve regenerate after trauma?]. PMID- 15343454 TI - [Neuroprotection and regeneration after traumatic lesion of the optic nerve]. AB - BACKGROUND: After a traumatic lesion of the optic nerve, retinal ganglion cells (RGC) undergo massive degeneration by apoptosis, which leads to loss of vision in the affected eye. Like other neurones in the central nervous system, RGC are not able to regenerate their damaged axons spontaneously. We used special surgical methods and pharmacological measures to achieve enhanced survival and regeneration of damaged RGC. MATERIALS AND METHODS: Studies were performed using the model of RGC degeneration induced by severing the optic nerve of adult rats. RGC were loaded with a fluorescent dye, and several drugs were applied intravitreally. The effects were evaluated after two weeks by counting the surviving RGC. For regeneration studies, an autologous peripheral nerve graft was sutured to the stump of the cut optic nerve, or the ends of the cut optic nerve were re-sutured. Recovery of RGC function was assessed by VEP measurements. RESULTS: The number of RGC surviving an axotomy increased significantly after intravitreal injections of aurintricarboxylic acid, cortisol, a caspase inhibitor, brimonidine or microglia-targeted substances. Regeneration of cut axons was enhanced by aurintricarboxylic acid or cortisol. In addition, considerable neuroprotective and regenerative effects including partial restoration of VEP were induced by lens injury, which results in a gradual release of crystallins into the vitreous, or by intravitreal injection of purified crystallins. CONCLUSION: The loss of vision after an optic nerve trauma can be reduced in this animal model by suitable neuroprotective measures, which raises hope for the treatment of patients. PMID- 15343455 TI - [Traumatic optic neuropathy--the present state]. AB - Direct or indirect trauma may damage the optic nerve and result in permanent visual loss. The diagnosis of traumatic optic neuropathy is not always straightforward and is complicated by unfavourable circumstances for the examination. The diagnosis should only by established if it can be based on a clear objective finding, a relative afferent pupillary defect or a pathological flash-evoked visual response. Concerning therapy, surgical decompression (even without direct injury to the nerve) or megadose steroids or both in combination have been recommended. But even spontaneous improvement may occur. A large multicentre study could not demonstrate a significant advantage for any of the measures recommended, not even against the spontaneous course. However, the study was not randomised. In animal experiments steroids have shown an unfavourable effect. Currently, therapeutic decisions have to be made on an individual basis, in the absence of any evidence-based recommendations. PMID- 15343456 TI - [Eye globe reconstruction after severe injury of the posterior segment]. AB - Severe injuries of the posterior eye segment mostly occur during perforation or rupture of the globe. The first treatment includes primary surgical repair of the ocular wound and prophylactic scleral buckling in case of involvement of the posterior segment. Thereafter, a vitrectomy should be performed to remove vitreous hemorrhage and intraocular foreign bodies and to achieve a stable attachment of the retina. Therefore, temporary perfluorocarbon and permanent silicone oil tamponade are used. A predictor of poor visual outcome is the presence of retinal and choroidal injury. In our patients, the most important prognostic factor was the initial visual acuity after the injury. Vitrectomy can significantly reduce the incidence of enucleation. Currently, better visual outcome is achieved by advances in surgical techniques. PMID- 15343457 TI - [Rupture of the choroid after eyeball contusion--an analysis based on the Erlangen Ocular Contusion Registry (EOCR)]. AB - BACKGROUND: Ocular injuries may lead to severe damage of the posterior segment with manifest visual impairment. Choroidal ruptures are frequently masked by acute subretinal haemorrhage. We analysed possible predictive factors and functional results of eyeballs with rupture of the choroid after ocular contusion. PATIENTS AND METHODS: We performed a retrospective study of 376 consecutive inpatients (Erlangen Ocular Contusion Registry - EOCR, over a 10-year period), who were treated because of a blunt eye injury at our eye hospital (86 % males). Detailed notes regarding the anterior and posterior segments were extracted from the standardised charts. Mean age was 28.8 +/- 16.1 years (4 to 84 years). Eyes with previous trauma or globe ruptures were excluded. RESULTS: Twenty-six of 376 patients developed choroidal rupture due to ocular contusion (6.9 %). A choroidal rupture was more frequent in females (9.4 %) than in males (6.5 %). Patients with choroidal rupture were treated as inpatients 5 days longer than patients without (10.7 vs. 5.5 days; p < 0.001). Twenty-two percent of the injuries occurred during work time. Main causes of choroidal ruptures were water jet (19 %), fireworks (12 %), elastic cords (12 %), metal pieces (12 %), gotcha (8 %) and champagne corks (8 %). The risk for developing a choroidal rupture due to water jet or fireworks injuries was increased 9 or 4 times. Ninety-two percent of choroidal ruptures were located at the posterior pole and concentric, 40 % were submacular, 12 % outside the large temporal vessels (4 % were located both centrally and peripherally). Initial visual acuity (VA) and VA at discharge were decreased significantly in eyes with rupture of the choroid (20/200 and 20/60) in contrast to eyes without (20/40 and 20/25; p < 0.001). Choroidal ruptures were often associated with iridodialysis, lens dislocation and contusion cataract (3 x ), vitreous haemorrhage (4 x ), complete retinal defects (6 x ), ciliary body clefts (7 x ) or hyphema rebleeding (4 x ). No association between the height of hyphema and choroidal ruptures was found. The predictive level of choroidal ruptures was 40 % in eyes with a combination of lens dislocation, traumatic cataract and vitreous bleeding. The final VA was 20/200 or less in 11 eyes associated with a prevalence of 55 % of ruptures submacularly. In contrast to this, eyes with VA > 20/200 developed 26 % submacular choroidal ruptures. CONCLUSIONS: Additional severe traumatic changes of the anterior and posterior segment were found 2 - 7 times more frequently in eyes with choroidal ruptures compared to eyes without those ruptures. The visual improvement was limited due to submacular ruptures. Frequent ophthalmological controls are recommended to minimise the risk of choroidal neovascularisation in a submacular location. PMID- 15343459 TI - Postcholecystectomy syndrome with special regard to children--a review. AB - The incidence of abdominal symptoms after cholecystectomy in adults is high. Up to one third of the patients consult a doctor because of abdominal complaints within 1 year after the operation. In addition, a higher incidence of colon carcinoma after cholecystectomy has been reported in female patients. This article reviews the known facts on the "postcholecystectomy syndrome". However, little is known about postcholecystectomy symptoms in children. Reports on cholecystectomy in children deal rather with feasibility than long-term outcome. Therapeutic concepts for children with symptomatic gallstone disease should consider the differences in the etiology of gallstone formation between children and adults. Therefore, we recommend a specific concept, including laparoscopic cholecystotomy, for children with temporary disorders causing gallstones, and laparoscopic cholecystectomy for all other patients. The impact of these procedures on postcholecystectomy symptoms in children and the impact of cholecystectomy during childhood on the incidence of right-sided colonic carcinoma remains to be determined. PMID- 15343460 TI - Surgery in biliary atresia--futile or futuristic? AB - Kasai's portoenterostomy is, so far, the only option for patients with biliary atresia (BA) to survive with their own liver. The long-term results are closely related to the timing of the procedure and to the experience of the center. However, optimal conditions cannot guarantee stable liver function. Unfortunately, the majority of patients with BA eventually need liver transplantation, making them the largest group of pediatric organ recipients. Thus, surgery in patients with BA treats only the symptoms, but never the cause of the disease. In order to focus on this point, international and interdisciplinary cooperation is mandatory to improve early and effective diagnosis, to optimize surgical therapy, and to coordinate clinical and basic research in BA. Uncovering its unknown aetiology is crucial for developing and modifying new therapeutic attempts to treat the disease, including the opportunities for prophylaxis. Until then, surgical treatment is still the best approach for biliary atresia and, so far, no further prospects are apparent. PMID- 15343461 TI - Parenteral nutrition-associated cholestasis--what do we know, what can we do? AB - In the early days of parenteral nutrition of children liver disease resulting in steatosis and cholestasis was assumed to be an inevitable complication of the procedure. Since then, the management of parenteral nutrition has improved so much that nowadays adolescents have a fair chance of surviving more than 15 to 20 years without severe liver disease. Nevertheless, we still see cases of parenteral nutrition-associated cholestasis (PNAC) due to various conditions such as recurrent infections, inflammatory response, inappropriate composition of the nutrient mixture, contaminants of the nutrient solution, and toxic substances from infusion bags and tubes. Recent research indicates that the administration of ursodesoxycholic acid and cysteine can prevent or even improve the cholestasis. A reversal of PNAC has been documented in an adolescent after small bowel transplantation from Japan. There is ample opportunity for prevention of PNAC with respect to the various pathophysiologic aspects: prevention, early detection, and management of infections, avoiding glucose overloads, cyclic infusion of nutrients, light protection of the solution, choice of paediatric amino acid solutions, and most important, oral or enteral feeding to support the bile flow by stimulating the cholecystokinine release. PMID- 15343462 TI - Conscious sedation: Off-label use of rectal S(+)-ketamine and midazolam for wound dressing changes in paediatric heat injuries. AB - BACKGROUND: Wound dressing changes after heat injuries expose the patient to repeated painful and frightening procedures in short intervals. Safe, adequate, and easily administered analgesia and sedation are required. The goal of this study was to evaluate the off-label use of rectally administered S(+)-ketamine and Midazolam by paediatric surgeons during repeated outpatient dressing changes for paediatric burns and scalding. PATIENTS AND METHODS: A total of 47 dressing changes of 30 children with I - IIa degrees burns were evaluated. Vital signs, side-effects, complications, anxiolysis, and analgesia were recorded during the procedure and for the following two hours. Patients were assessed by a discharge scoring system and an age-appropriate pain scoring system at regular intervals. Before discharge, parents were interviewed on their level of satisfaction with the protocol. RESULTS: Adequate sedation and analgesia was achieved in 44 procedures (94 %). No complications and, in particular, no compromise of breathing, ventilatory, and cardiovascular functions were recorded. The discharge scoring system indicated a return to baseline function 30 minutes after the procedure in all patients. The parents were generally very satisfied with the protocol. All children old enough to be questioned were found to have an anterograde amnesia for the duration of the procedure. CONCLUSION: Conscious sedation with rectally applied S(+)-ketamine and Midazolam allows safe and painless dressing changes after heat injuries in children. PMID- 15343463 TI - Hyperbaric oxygenation therapy for simple adhesive postoperative intestinal obstruction in children: comparison of the use of a short tube versus a long tube. AB - We hypothesized that hyperbaric oxygenation (HBO) combined with a long tube (LT) [HBO + LT] would be more effective than HBO combined with a short tube (ST) [HBO + ST] for simple adhesive postoperative intestinal obstruction (APIO) in children, assuming that there is synergism between HBO and LT. The objective of this study was to determine retrospectively the effect of HBO + LT compared to HBO + ST for simple APIO in children. Seventy-three patients were diagnosed with simple APIO, and 51 of these patients were treated with HBO + LT during 104 HBO sessions, while 22 were treated with HBO + ST during 34 HBO sessions. HBO was performed at a pressure of 2 atmospheres for 60 minutes once daily. The recovery rates after HBO therapy were 87.5 % for the HBO + LT group and 82.4 % for the HBO + ST group (p = 0.4496). HBO was performed 5 +/- 3 (range 1 to 15) times for the HBO + LT group and 4 +/- 2 (range 1 to 8) times for the HBO + ST group (p = 0.9847) for ultimate recovery from simple APIO. The recovery rate after HBO therapy of up to 6 sessions was 78.2 % for HBO + LT and 92.1 % for HBO + ST (p = 0.0360) among the cases that recovered ultimately. The effect of HBO + LT did not significantly differ from that of HBO + ST, but the insertion of an LT is more intricate and the cost is higher than that of an ST. Therefore, we conclude that the use of an LT is not necessarily required for HBO therapy for simple APIO. PMID- 15343464 TI - Distal catheter obstruction from non-infectious cause in ventriculo-peritoneal shunted children. AB - In hydrocephalic children, ventriculo-peritoneal shunting is the preferred treatment with few complications. However, an obviously non-infectious peritoneal reaction to the cerebrospinal fluid (CSF) may occasionally lead to shunt malfunction. In eight hydrocephalic children, shunt malfunction with distal catheter complication was found with abdominal pseudocyst formation in seven cases and accumulation of the CSF in one. All children had a normal CSF cell count and glucose concentration, and white cell count, and C-reactive protein in peripheral blood were normal. No CSF infection could be detected despite prolonged aerobic and anaerobic cultures. After initial externalisation of the shunt and subsequent routine administration of antibiotics because infection initially was suspected, ventriculo-peritoneal shunting was attempted one to three times with identical failure before successful conversion to a ventriculo atrial system. At laparotomy the peritoneum and intestinal serosa were hyperaemic and oedematous in all patients, five of whom also had pseudocysts and two of whom also had intra-abdominal adhesions. Four children had a revision 6-24 years after the ventriculo-atrial conversion due to short atrial catheter with distal obstruction. In three of them, the distal catheter was successfully replaced into the peritoneal cavity. The fourth child, however, developed an infectious abdominal pseudocyst with adhesions due to a then undetected Propionibacterium acnes infection. After externalisation and antibiotics, a new ventriculo-atrial shunt was inserted. At follow-up between 5 months to nearly 6 years later, the three children with peritoneal catheters did not show any signs of shunt malfunction or abdominal problems. Thus hydrocephalic children may develop shunt malfunction with distal catheter obstruction due to a still unexplained, transient, non-infectious peritoneal reaction leading to abdominal pseudocyst formation or accumulation of CSF. In some children, however, it may later be possible to replace the distal catheter into the peritoneal cavity, if no infection is involved. PMID- 15343465 TI - Comparison of closed-tube thoracostomy and open thoracotomy procedures in the management of thoracic empyema in childhood. AB - PURPOSE: There is still an argument concerning the correct management of pleural empyema. The aim of this study is to compare the effectiveness of closed-tube thoracostomy and open thoracotomy procedures in the management of empyema in children. METHODS: This is a prospective study of 30 patients with parapneumonic empyema who were managed randomly either by closed-tube thoracostomy or open thoracotomy procedures. The two procedures were compared based on the respective times to achieving normal body temperature and breath rates, duration of tube drainage, length of hospitalization, and complication rates. Both groups were also assessed by comparing tube drainage duration, pleural fluid pH, agent pathogen and glucose level. RESULTS: Average tube duration was 7.5 +/- 1.1 days and average hospital stay was 9.5 +/- 1.5 days in the open thoracotomy group. In the closed tube thoracostomy group tube, duration was 13.8 +/- 2.3 days and average hospital stay 15.4 +/- 2.3 days. In the open thoracotomy group 73.3% of the patients had achieved normal body temperature and 66.7 % had a normal breath rate within the first 48 hours. In the closed tube thoracostomy group these rates were 40% and 20%, respectively. In both groups, tube drainage duration was found to be longer in patients whose pleural fluid pH was < 7.2. CONCLUSION: The authors conclude that open thoracotomy is a safe, efficient, and easy method of treatment for pleural empyema in children. It was also observed that pleural fluid pH level is the most important prognostic criteria in pleural empyema. PMID- 15343466 TI - Surgical treatment of pulmonary hydatid cysts in children: report of 66 cases. AB - AIM OF THE STUDY: Hydatid disease is a parasitic infestation which is endemic in many sheep and cattle raising areas and is still an important health problem in the world. The aim of our study was to present our surgical experience and strategy in the management of pulmonary hydatid disease. METHODS: Sixty-six patients with 83 pulmonary hydatid cysts underwent surgical treatment in our institution between January 1990 and March 2003. We used double-lumen endotracheal tubes in children older than 12 years who were operated on for hydatic cyst in the last 8 years. RESULTS: There were 38 boys and 28 girls with a mean age of 9.6 +/- 7 years (range 5-15 years). Of the 83 cysts, 61 were intact and 22 were ruptured cysts. Isolated pulmonary hydatid cyst was seen in 61 patients (92.4 %), while 5 patients (7.6 %) had combined pulmonary and hepatic cysts. Lateral thoracotomy was performed in 54 patients (82 %), thoracophrenotomy in 5, bilateral thoracotomy in 4, and median sternotomy in 3 patients. Cystotomy and capitonnage was performed in 58 cysts, cystotomy alone in 21, and resection techniques were used in 4. There were 8 postoperative complications in 7 patients. The most common complication was atelectasis. CONCLUSIONS: Surgery is the treatment of choice for most patients with pulmonary hydatid cysts. The usage of double-lumen tubes may decrease intra- and postoperative complications. Thoracophrenotomy can be chosen as the surgical procedure in the management of hepatic and pulmonary hydatid cysts. PMID- 15343467 TI - Appendicitis in children--impact of US and CT on the negative appendectomy rate. AB - BACKGROUND/PURPOSE: Ultrasonography (US) and computed tomography (CT) have proved useful for the evaluation of suspected appendicitis and were introduced as diagnostic tools at our institution about ten and five years ago, respectively. The aim of this study was to evaluate how the negative appendectomy and perforation rates have changed with increased use of US and CT. METHODS: The medical records of 600 children who underwent appendectomy during years 1991, 1994, 1997, and 2000 were reviewed. Perforation in perforated appendicitis was considered to have occurred after admission if the time interval between the first health professional contact and surgery exceeded 12 hours. RESULTS: The total number of appendectomies during the years 1991, 1994, 1997, and 2000 was 406, 334, 407, and 397, respectively. The negative appendectomy rate for the same years was 23%, 8.7%, 8.0%, and 4.0%, respectively. The overall rate of perforations and the perforation rate after admission was 32% and 12%, 34% and 7.3%, 34% and 13%, and 29% and 2.1%, respectively. The rate of patients who underwent US and CT during each period was 1.0% and 0.0%, 41% and 0.0%, 91% and 21% and 98% and 59%, respectively. CONCLUSIONS: The negative appendectomy rate has been substantially reduced after the introduction of both US and CT. The rate of perforation after admission has not increased. PMID- 15343468 TI - Intra-abdominal, retroperitoneal, and visceral abscesses in children. AB - This review describes the microbiology, diagnosis, and management of intra abdominal abscesses (including subphrenic, hepatic, splenic, and retroperitoneal abscesses) in children. They often occur as a complication of local or generalized peritonitis, commonly secondary to appendicitis, necrotizing enterocolitis, pelvic inflammatory disease, and tubo-ovarian infection, surgery or trauma. The original infection generally occurs because of the entry of enteric microorganisms into the peritoneal cavity through a defect in the wall of the intestine or other viscus as a result of obstruction, infarction, or direct trauma. Mixed aerobic and anaerobic flora can be recovered from most abscesses. The predominant aerobic isolates are Escherichia coli, Staphylococcus aureus, and Enterococcus spp. and the main anaerobic bacteria are Bacteroides fragilis group Peptostreptococcus spp., Fusobacterium spp., and Clostridium spp. The treatment of intraabdominal abscesses includes drainage, surgical correction of pathology, and administration of antimicrobials effective against both aerobic and anaerobic microorganisms. PMID- 15343469 TI - Tracheoesophageal fistula due to disc-battery ingestion. AB - Ingestion of a disc battery can lead to hazardous complications including tracheoesophageal fistula (TEF), especially when the battery is impacted in the esophagus. Urgent esophagoscopic removal of the battery is essential in all cases. Persisting respiratory symptoms and/or feeding difficulty after removal should alert the surgeon to evaluate for TEF. Esophagography and/or esophagoscopy should be performed. Once the TEF is identified, conservative management is the initial treatment of choice. Delayed primary repair can be tried if spontaneous closure does not occur. The authors present a case of TEF secondary to disc battery ingestion with review of the current literature and discuss the management of this rare but potentially life-threatening complication. PMID- 15343470 TI - Uncommon case of a foreign body ingestion with consecutive small-bowel obstruction in a child. AB - A 12-year-old boy with Lennox syndrome presented with an acute abdomen and a history of progressive abdominal pain and vomiting over 3 weeks. The uncommon finding in this case was a foreign body detected in a lower loop of the jejunum causing radiological and clinical signs of jejunitis/ileitis. The foreign body had to be removed surgically and turned out to be a hard (originally soft) plastic part of a towel rack. PMID- 15343471 TI - Congenital esophageal stenosis: a case report and review of the literature. AB - A child with congenital esophageal stenosis (CES) demonstrates the need for accuracy in the diagnosis and management of this rare problem. Other etiologies of dysphagia must be looked into, but when the expected results are not realized CES should be considered. A range of diagnoses and diagnostic studies may help to get children with dysphagia the treatment that they need to resolve their symptoms. PMID- 15343472 TI - Post-traumatic duodenopancreatectomy in a child. AB - We describe a 5-year-old boy who complained of severe abdominal pain after a car accident. He was sitting on the lap of the driver and his abdomen was pressed closely between the pregnant driver and the steering wheel. At operation, associated injuries to the duodenum and pancreas were detected which could not be repaired. The transected pancreas and extensive duodenal injury underwent a Whipple-type reconstruction. On postoperative day six, the patient was re operated due to extensive liver necrosis which was detected on abdominal computerized tomography and a partial liver resection was performed. He was discharged on day 16 after the second operation and has done well so far. PMID- 15343473 TI - Imperforate anus associated with atresia of the transverse colon: a case report. AB - We describe here a rare case of association of imperforate anus with transverse colon atresia in a male neonate. Preoperative X-ray studies demonstrated: a) a distended loop in the epigastrium with fluid levels on plain A/P upright radiography, b) absence of pelvic intestinal gas on the invertogram 16 hours later. A three-stage operative approach was undertaken comprising resection of the atretic loop and colostomy, posterior sagittal anorectoplasty a few months later, and finally closure of the colostomy. The postoperative outcome was good. The association of these anomalies should be kept in mind in neonates with anal atresia and abdominal distention. PMID- 15343475 TI - [Psychotherapy within groups]. PMID- 15343476 TI - [Differential group experiences of cognitive-behavioral and psychodynamic group psychotherapy]. AB - Research concerning the question, whether and to what extent cognitive-behavioral (CB) and psychodynamic (PD) therapy consist of differing process components under clinical representative conditions, is relevant especially for a valid interpretation of comparative outcome research, for identifying differential beneficial factors of psychotherapy and for a systematic indication for, respectively assignment of, patients to the two treatments. In this study it is investigated whether PD and CB differ concerning the realisation of factors of group experience, respectively of beneficial group elements (e. g. cohesion, catharsis, learning by feedback). For this purpose, in a naturalistic design, a stratified sample (N = 36) of 104 videotaped sessions (PD groups, interactional CB groups and indicative CB groups; N = 171 patients with a broad spectrum of F diagnoses of ICD-10, especially F3/F4) were rated by observers using the Kieler Gruppenpsychotherapie-Prozess-Skala (KGPPS). Analyses of variance and a priori Helmert-contrasts reveal differences between PD and CB with at least medium effect sizes in 12 of the 16 factors of group experience. However, differences also were found between the two CB group treatments (9 factors of group experience with differences with large effect sizes). The results suggest that the different treatment approaches foster different qualities and quantities of group experience and that the latter seems not to evolve from the group context "per se" (i. e. by the plurality of the group). PMID- 15343477 TI - [Quality of life in patients with malignant glioma and their relatives]. AB - Maintaining Quality of Life (QoL) plays an important role in oncology as despite an increasing number of therapeutic opportunities numerous tumours can not be cured yet. Among those tumours are malignant gliomas. Their prognosis is usually very poor. For patients and their relatives the diagnosis of this life threatening disease is a crucial moment in their lives which may be followed by significant decrease of QoL due to both the poor prognosis and the existing or increasing neurological deficits. The central question of this study was to determine whether the disease has an impact on the QoL of patients and their relatives and what kind of impact that may be. Therefore, QoL was investigated in patients currently being treated as well as in long-term survivors. The results indicate that the diagnosis of a malignant glioma confronts both the patients and the relatives with a disease process that predisposes to restrictions in QoL but does not lead necessarily to a poor QoL. For this reason, only the evaluation of individual circumstances and strains allows to assess QoL and hence specific interventions to maintain or re-establish QoL. PMID- 15343478 TI - [The measurement of the depersonalisation-derealisation-syndrome with the German version of the Cambridge Depersonalisation Scale (CDS)]. AB - Self-rating scales have proved to be essential in the study of depersonalisation, which regrettably, is still seldom recognised in clinical practice. In recent studies the Cambridge Depersonalisation Scale (CDS) has emerged as an useful instrument for the study of depersonalisation. Here we report a validation study of the authorised German version of the CDS in a sample of 91 inpatients, 43 of whom had pathological depersonalisation and 48 without pathological depersonalisation. The SCID-D Interview for depersonalisation and derealisation was used as the gold standard and the German version of the Dissociative Experiences Scale was used to test the external validity of the scale. The German version of the CDS was found to have high internal consistency and reliability (alpha = 0,95 and Guttman Split-half = 0,95) and could differentiate patients with pathological depersonalisation from the control group. We therefore conclude that the German version of the CDS can be considered as reliable and valid. PMID- 15343479 TI - [The Adult Attachment Scale (AAS) - psychometric evaluation and normation of the German version]. AB - Based on conceptualizations and assessment instruments of adult attachment research, the development of the adult attachment scale is outlined. The AAS scale is a self report instrument reflecting attachment-related attitudes. The dimensional scales of the AAS assess to what extent a person feels close to his partner, to what extent he is able to rely on others, and how much someone is afraid of losing an intimate partner. The German version of the AAS was normed in a representative study and its psychometric properties were investigated. The reliability of all scales was satisfactory (alpha = 0.72 - 0.79), however, the scale structure only acceptable. Thus, the current version might be integrated in studies to analyze the convergence of adult attachment measures. Findings on the convergent and discriminant validity of the scales (sociodemographic data, the SOC-S, the HADS-D, the SOMS and the Whiteley-Index were included) suggest a high prognostic relevance of the scales in the fields of health psychology and psychosomatics. PMID- 15343480 TI - [Religious dimensions and optimistic perception of the future in a sample of students]. AB - Religiosity and the less circumscribed construct of spirituality may be related to resilience and salutogenesis. Most inventories focus more on religious practice, as prayer or visiting church, than systems of belief. The relationship between "existential well-being" and "religious well-being" has been investigated in a sample of young students by using the "Religious" and the "Existential Well Being Scale" (Ellison u. Paloutzian 1982). Optimistic perception of the own future was unrelated to any of the religious well-being items. PMID- 15343481 TI - [The convergence between nature and mind does not exist because nature is mindful but because the mind is natural]. PMID- 15343482 TI - [Alpha-1-antitrypsin-laboratory and German alpha-1-antitrypsin-registry]. PMID- 15343483 TI - [The pneumologist as detective]. PMID- 15343484 TI - [Autofluorescence bronchoscopy]. PMID- 15343485 TI - [Autofluorescence as routine? -- pro]. PMID- 15343486 TI - [Autofluorescence bronchoscopy -- ready for routine? -- contra]. PMID- 15343487 TI - [Quality of life before and during nCPAP]. AB - The obstructive sleep apnea syndrome is accompanied by an impairment of patients' condition during the day. The latter, in turn, is a decisive criterion quality of life. The instruments employed for data gathering, objectification and observation of the course of therapy are questionnaires. Questionnaires like Satisfaction With Life Scale SWLS: for global quality of life; Munchner Lebensqualitats-Dimensionen-Liste MLDL and Short Form-36 of the Health Survey SF 36: for specific areas of life quality are often used. The purpose of this study is to clarify, to what extent and in which areas quality of life of OSAS patients improves both after initiation of therapy and under long-term therapy. By comparing effect size the value for effectiveness of therapy can be estimated. Thus, 41 OSAS patients and 40 controls were given an extensive set of tests addressing parameters of both sleep and quality of life. Testing of global quality of life with the Satisfaction With Life Scale (SWLS) did not reveal any significant difference between the controls and the OSAS patients. In specific araeas of life quality, 2 (of 4) subscales (Physical satisfaction: p = 0.003; Psychological satisfaction: p = 0.001) of the Munchner Lebensqualitats Dimensionen-Liste (MLDL) showed differences between the controls and the OSAS patients. For the Short Form-36 of the Health Survey (SF-36), 7 (of 8) subscales indicated significant differences between this groups (except for Emotional Role Function). The comparison of the effect size revealed the strongest effect in the subscale vitality of the SF-36 after more than 6 months (0.93). Good effect sizes were found in the domains Physical satisfaction (0.70) and Psychological satisfaction of the MLDL (0.56). In conclusion the MLDL is in consideration of its simple performance and evaluation compared with the SF-36 the better instrument in the clinical routine for the coverage of improved Quality of Life through a nCPAP therapy. PMID- 15343488 TI - [Extramedullary hematopoiesis -- a differential diagnosis of intrathoracic paravertebral tumors]. AB - BACKGROUND: Intrathoracic tumors always suggest a malignant clinical picture in the first instance. Especially in the case of paravertebral masses an extramedullary hematopoiesis is possible. It is regarded as compensation mechanism for reduced blood cell formation. PATIENTS: In four patients' radiographic examination of the lung partially bilateral paravertebral masses where detected by chance. In two cases, a chronic anemia existed, one patient suffered from an extended hemangiomatosis of the right brachium and the right hemithorax. In one case, no hematological disease could be diagnosed. RESULTS: The computed tomography revealed malignancy-simulating tumors with partially necrotic pattern. In all cases, transthoracal biopsy showed cytological evidence of typical differentiated bone marrow with a regular hematopoiesis. As the extramedullary hematopoiesis of these four patients was diagnosed by chance and no symptoms or local complications existed, no specific therapy was necessary. In one of these cases a stable radiographic outcome for more than ten years is evident. CONCLUSIONS: Especially in the case of patients with chronic anemia the existence of extramedullary hematopoiesis has to be considered upon occurrence of paravertebral tumors. An hematological disorder as cause may as well be absent. Due to the suspicion of malignancy a morphologic diagnosis is always necessary, but some clinical and radiologic features may indicate a benign process. A progression of the size can be unrecognized for ten years, so a therapeutic intervention is needed only in case of symptoms like myelocompression. PMID- 15343489 TI - [Misting-fountain-alveolitis]. AB - A 22-year-old woman developed recurrent episodes of fever, cough and dyspnea after repeated exposure to a misting fountain at home. A diagnosis of extrinsic allergic alveolitis (EAA) was made by detection of serum antibodies against the fountain water, by culture of Bacillus subtilis, Mucor racemosus, Mucor mucedo, and Saccharomyces cerevisiae from the water, and by detection of specific IgG antibodies against Bacillus subtilis and the Mucores. The diagnosis was confirmed by a restrictive lung function pattern, and a highly increased total cell count with a lymphocytosis of 39 % in the bronchoalveolar lavage. An inhalation challenge with the misting fountain resulted in a positive reaction. Because this humidifier system has recently become widespread at home, clinicians should be aware of this specific type of EAA which may be called "misting fountain alveolitis". PMID- 15343490 TI - [Pleuromesothelioma -- psychology and pathogenesis]. AB - Investigations into the anatomical pathology are of crucial importance for the diagnosis and therapy of pleural tumours. The main objective is to differentiate reactive hyperplastic proliferations of the mesothelium from already manifest primary or secondary malignant pleural tumours. Given the absence of distinct morphological features displayed by pleural mesotheliomas, specific additional immunohistochemical and cytometric tests can provide valuable clues for the differential diagnosis of primary and secondary pleural neoplasms. Confirmation of the diagnosis during the initial stages of tumour development i. e. mesothelioma in-situ and early mesothelioma necessitates invasive diagnostic procedure as early as possible. This holds particularly true if surgery is aimed at cure. However, in order to firmly establish the diagnosis considering the afore mentioned problems, the tissue specimens to be examined by a pathologist must be of adequate size. 90 % of all pleural mesotheliomas are asbestos associated due to occupational exposure. Therefore, all cases of a suspected pleural mesothelioma must be reported to the appropriate professional association as a potential occupational disease. The Bochum based German Mesothelioma Registry, which is supported by the head office of the German Professional Associations, conducts research and acts as an advisory institution in difficult cases, where problems concerning the classification of tumours by their anatomical pathology arise. PMID- 15343491 TI - [The history of bronchial carcinoma]. AB - Up to the mid 19 (th) century primary bronchial carcinoma was unknown. Primary tumours and metastases of malignant lung tumours were often not distinguished one from the other. Only in 1871, Theodor Langhans from Marburg reported the first certain observation of bronchial carcinoma. Definite information on the increasing frequency of lung and bronchial carcinomas at the beginning of the 20 (th) century was only obtained by autopsy statistics. The Cancer Registry of the Saarland showed that the incidence of lung cancer has not increased since 1970 in men, but has tripled in women. Already in 1923, Theodor Fahr from Hamburg referred to the danger of smoking. In Germany the so-called Schneeberg's lung cancer plays an important role among occupational lung cancer diseases. Only after Germany's reunification the drama of the uranium miners of the former so called Wismut AG became fully known. Regarding occupational diseases, asbestos related bronchial carcinomas and mesotheliomas are at the top of the causes of death today. PMID- 15343492 TI - Are nurses at the table? A new nursing degree could help. PMID- 15343493 TI - Get out of town! PMID- 15343494 TI - Lost in translation? PMID- 15343495 TI - The clinical partnership as strategic alliance. AB - The purpose of this article is to describe a renewed partnership between a collegiate school of nursing and a community hospital. Universities and hospitals are searching for creative solutions to increase the number of registered nurses available to meet the demand for nursing care. An affiliation agreement had been in existence for many years, but health care system imperatives made it necessary to redesign the partnership between nursing education and nursing service. The model used to develop this new partnership is based on the work done in the field of management and is in the form of a strategic alliance. The success of a strategic alliance depends on two key factors: the relationship between partners and partnership performance. Identified outcomes show that this partnership is helping to meet the increasing demand for nursing care by building student capacity, satisfying mutual needs of faculty and clinical staff, and removing economic barriers. This article describes the development of the strategic alliance, its current status, and strategies for the future. PMID- 15343496 TI - Genetic discoveries and nursing implications for complex disease prevention and management. AB - The purpose of this article is to examine the management of patients with complex diseases, in light of recent genetic discoveries, and to explore how these genetic discoveries will impact nursing practice and nursing research. The nursing science processes discussed are not comprehensive of all nursing practice but, instead, are concentrated in areas where genetics will have the greatest influence. Advances in genetic science will revolutionize our approach to patients and to health care in the prevention, diagnosis, and treatment of disease, raising many issues for nursing research and practice. As the scope of genetics expands to encompass multifactorial disease processes, a continuing reexamination of the knowledge base is required for nursing practice, with incorporation of genetic knowledge into the repertoire of every nurse, and with advanced knowledge for nurses who select specialty roles in the genetics area. This article explores the impact of this revolution on nursing science and practice as well as the opportunities for nursing science and practice to participate fully in this revolution. Because of the high proportion of the population at risk for complex diseases and because nurses are occupied every day in the prevention, assessment, treatment, and therapeutic intervention of patients with such diseases in practice and research, there is great opportunity for nurses to improve health care through the application (nursing practice) and discovery (nursing research) of genetic knowledge. PMID- 15343497 TI - Evaluation of computerized nursing care plan: instrument development. AB - With the increasingly popular use of information technology in patient care, the need for reliable instrumentation to evaluate information systems has become critical. This article describes the psychometric testing of a scale developed to evaluate a computerized nursing care plan (CNCP) system. A review of the literature generated a 44-item questionnaire, which was then administered to a convenience sample of 729 hospital nurses in Taiwan. Factor analysis (principal component analysis with varimax rotation) and item analysis were applied to establish the scale's construct validity and reliability. Twenty-two items selected from the original 44-item pool were grouped into 6 major constructs: patient care, nursing efficiency, professionalism, usage benefit, education and training, and usability. The alpha coefficient was 0.85. The statistical results showed that nurses generally valued using the CNCP system. Further psychometric analysis of the scale is suggested in other nursing populations, for subscale development and to refine item wording. PMID- 15343498 TI - Research on faculty orientation programs: guidelines and directions for nurse educators. AB - The objective of this study was to review the literature for the quality of evidence available regarding faculty orientation programs and to identify practice and research implications. Computerized searches in Medline, Cumulative Index to Nursing and Allied Health Literature, and Educational Resources Information Center, and references cited in articles, were the data sources reviewed. Keywords used in the search were faculty orientation, faculty development, faculty development programs, faculty mentoring, new faculty needs, nursing, teaching, and mentoring. All relevant articles published after 1980 were evaluated. Nineteen reports of research from indexed journals in English relevant to the keywords were reviewed: 14 were quantitative reports, and 5 were qualitative. Characteristics consistently present in the literature were that an orientation program takes place over a prolonged period of time, is incorporated into faculty development plans, creates or fosters an inviting environment, includes information about the tripartite role of the academician, and identifies a specific institutional resource person such as a mentor. Most research to date has been descriptive, consistent with the third level of quality of evidence (III; Marek, 1995). Faculty for whom orientation programs are offered might immerse themselves more effectively in their new environments. Mentoring relationships can ease faculty transitions. PMID- 15343499 TI - International rural health needs and services research: a nursing and midwifery response. AB - As the importance of evidence-based practice in global health care increases, the need for constant, accurate feedback from those in practice is critical, yet the requirement to rapidly obtain and disseminate data on a global basis is a challenge for all health professionals. The Internet allows for global participation in data collection that dramatically streamlines the traditional survey process. This new paradigm applies to surveys that are short, issue focused, and time sensitive. The Global Network of World Health Organization (WHO) Collaborating Centres for Nursing and Midwifery Development used a sample of international nurses in 70 countries to identify the worldwide rural health issues of WHO priority health needs, treatment modalities, health care interventions, and providers, as well as the current state of rural health research on a global level. This article presents the results of the survey regarding rural health needs and nursing and midwifery's response to them internationally, demonstrates Internet data collection, and shows how this research paradigm can help establish an evidence base for nursing practice. PMID- 15343500 TI - The Nurse Reinvestment Act: the impact of governmental and nongovernmental administrative tools. AB - The Nurse Reinvestment Act has ushered in a new wave of federal involvement intended to address the issue of the critical nursing shortage. To implement this act, administrators will rely on a series of governance tools often called upon to address issues related to the social welfare of the country. Included among these tools are grants and loans through which the intent of the legislation will be formalized. The purpose of this analysis is to provide an overview of the characteristics associated with these tools, specifically as they relate to nursing education. Consideration will be given to the political environment through which funding, and thus the ability to implement the Nurse Reinvestment Act, will be realized, and the potential obstacles when executing federal programs in a market-driven economy. PMID- 15343504 TI - Phyllodes tumor of the breast: local recurrence versus metastatic capacity. PMID- 15343505 TI - Stromal expression of vascular endothelial growth factor correlates with tumor grade and microvessel density in mammary phyllodes tumors: a multicenter study of 185 cases. AB - A retrospective review of 185 mammary phyllodes tumors (105 benign, 51 borderline, 29 malignant) from 4 centers was performed by immunohistochemistry to investigate the expression of vascular endothelial growth factor in the epithelial and stromal cells of mammary phyllodes tumors. The correlation of vascular endothelial growth factor with tumor grade, stromal cell nuclear pleomorphism, cellularity, mitotic rate, margin histomorphology, and the stromal microvessel density was evaluated. Vascular endothelial growth factor expression was found in the epithelium in 29% and in the stromal cells in 31% of cases. There was significant increase of vascular endothelial growth factor expression in the stromal cells with increasing degree of malignancy, but not the epithelium. Microvessel density in the stroma also showed significant correlation with tumor malignancy, and a correlation was shown with the stromal vascular endothelial growth factor expression. Statistical overlap of stromal vascular endothelial growth factor and microvessel density in predicting malignancy suggests that angiogenesis may be an effector mechanism for vascular endothelial growth factor. Assessment of stromal VEGF may be useful as an adjunctive diagnostic criterion in the histologic assessment of malignancy in phyllodes tumors. PMID- 15343506 TI - The effects of glyoxal fixation on the histological evaluation of breast specimens. AB - Glyoxal (GL), a non-formalin-containing aldehyde tissue fixative, is advocated as a superior fixative that is environmentally safe and lacks the purported carcinogenic health hazards associated with formalin use. In addition, it is advertised as requiring no antigen retrieval before immunohistochemical staining. We compared GL fixation to standard formalin fixation of breast specimens removed for microcalcifications or breast tumors. Although the hematoxylin and eosin morphology of GL-fixed and formalin-fixed tissues was equivalent, detection of microcalcifications in GL-fixed breast specimens was hampered by loss of basophilia, likely due to increased calcium solubility in glyoxal. Moreover, estrogen receptor detection in GL-fixed specimens was diminished compared to formalin and did require antigen retrieval. PMID- 15343507 TI - Molecular cytogenetic characterization of recurrent translocation breakpoints in bizarre parosteal osteochondromatous proliferation (Nora's lesion). AB - Bizarre parosteal osteochondromatous proliferation (BPOP), or Nora's lesion, is a rare tumorous lesion with aggressive growth that affects primarily the small tubular bones in the distal extremities and often recurs after excision. No previous cytogenetic data on BPOP are available. In the present study, lesions from 5 patients were investigated by chromosome banding and fluorescence in situ hybridization (FISH) analyses. Patient age ranged from 24 to 46 years, and the lesions were located in the fingers in 4 cases and in a toe in 1 case. Histological sections from all 5 tumors were characterized by a mixture of hypercellular cartilage, cancellous bone, and spindle cell components. Samples from 2 patients were available for cytogenetic analysis. One of these showed a normal female karyotype, and the other revealed a balanced translocation, t(1;17)(q32;q21), as the sole anomaly. The translocation was further characterized by 3-color metaphase FISH analyses, using 17 1q32-specific and 18 17q21-specific bacterial artificial chromosome probes, to map the precise location of the breakpoints. Split signals were detected by the RP11-99A19 probe in chromosome 1 and by the RP11-219F9 probe in chromosome 17. To determine whether these rearrangements are characteristic features of BPOP, paraffin embedded tissue sections from all 5 patients were investigated by interphase FISH analyses. All 5 cases had a break in 1q32, and 4 of the 5 cases showed a break in the 17q21 region. The results strongly indicate that t(1;17)(q32;q21), or variant translocations involving 1q32, are recurrent and unique aberrations in BPOP. Several genes are located within the 2 sequences spanning the breakpoints, and further studies should be performed to determine whether any of these are involved in the formation of a fusion gene. PMID- 15343508 TI - Nonalcoholic steatohepatitis: histologic features and clinical correlations with 30 blinded biopsy specimens. AB - Thirty overweight patients with clinically characterized and biopsy proven nonalcoholic steatohepatitis (NASH) were enrolled in a 48-week treatment trial with rosiglitazone, a peroxisome proliferator-activator receptor (PPAR)-gamma agonist that enhances insulin sensitivity. Improvement in laboratory liver tests, insulin resistance and liver fat content were documented; blinded biopsy review demonstrated decreases in necroinflammatory activity or grade and in individual components of grade, and changes in the relationship of lobular and portal inflammation as well as in the nature of perisinusoidal fibrosis. The current study identified correlations of histological features of the protocol entry biopsy specimens with contemporaneous laboratory and imaging tests. Significant correlations with histologically assessed steatosis were liver fat, evaluated by computed tomography (P = 0.001); mean HbA1C, a measure of glycemic control (P = 0.004); and QUICKI, a measure of insulin sensitivity (P = 0.05). Histologically determined grades of steatohepatitis (SH) correlated with HbA1C (P = 0.01), and a trend toward elevated fasting glucose levels was seen. No subject in the study was cirrhotic at entry; fibrosis scores of the 30 subjects did not significantly correlate with age, gender, body mass index, or clinical tests. All subjects underwent 3 biopsies (prior, entry, and posttreatment), and all had undergone a prior biopsy with diagnostic SH. By blinded analysis, 7 study entry biopsy specimens did not fulfill published strict criteria for SH. Laboratory results from these subjects included normal fasting glucose level and, compared with the 23 subjects with criteria for SH, lower mean alanine aminotransferase and aspartate aminotransferase levels (P = 0.02 for both), less insulin resistance (P = 0.03), and lower mean HbA1C (P = 0.001). We conclude that biopsy findings determined by blinded analysis correlated with image-detected steatosis, laboratory markers of hepatic inflammation, insulin resistance, and long-term glycemia; the findings confirm the usefulness of strict histological criteria in the evaluation of NASH. PMID- 15343509 TI - The role of liver biopsy in the management of chronic hepatitis C in patients infected with the human immunodeficiency virus. AB - We evaluated the role of liver biopsy in the management of chronic hepatitis C in HIV-infected persons. Patients included had abnormal alanine transaminase (ALT) levels, detectable HCV RNA, and an interpretable liver biopsy. Demographic, epidemiologic, clinical, laboratory, histological, and therapeutic data were recorded in all patients. We also registered the clinical diagnosis of cirrhosis (previous to biopsy) and whether the biopsy result deferred the decision of initiating therapy. During the 33-month duration of the study, 112 patients were included. The degree of fibrosis in liver biopsies was none or mild (F0 or F1) in 47 patients (42%) and was significant or severe in 65 (58%). Seventeen patients (15%) had histological cirrhosis. By logistic regression analysis, only portal hypertension (odds ratio [95% confidence interval], 5.3 [1.05-25.9], P = 0.04) was independently associated with significant fibrosis. Overall, cirrhosis was predicted before biopsy in 29 patients (26%) by the caregiving physician, but only 8 of these were confirmed histologically. The clinical prediction of cirrhosis before biopsy had a sensitivity of 47%, a specificity of 78%, a positive predictive value of 28%, and a negative predictive value of 89%. Histological findings changed the decision to initiate HCV therapy in 19 patients (17%) because of little or no fibrosis. Liver biopsy is a useful tool in the management of HCV-HIV-coinfected persons. In addition to allowing grading and staging of the disease far better than any other method or combination of methods, it is important for making management decisions for patients coinfected with HCV and HIV. PMID- 15343510 TI - Fibrosis is worse in HIV-HCV patients with low-level immunodepression referred for HCV treatment than in HCV-matched patients. AB - Hepatitis C virus (HCV) infection is frequent in human immunodeficiency virus (HIV)-infected patients. It is known to have an aggressive course in significantly immunosuppressed patients, and cirrhosis C has become one of the main causes of mortality in HIV-HCV coinfected patients since the improvement of antiretroviral therapy. The reasons for this severe fibrotic evolution are unclear. This prospective study compared chronic HCV lesions, liver immunocompetent cells, fibrosis and liver HCV loads in 2 cohorts of naive patients referred for HCV treatment: 33 HIV-HCV coinfected patients with CD4 >250/microL and 33 HCV-infected patients matched for the main risk factors of fibrosis. Fibrosis, particularly perisinusoidal fibrosis, was more marked in the coinfected patients. This occurred in the absence of a significant difference in disease activity. The number of CD3+ cells in the liver was higher in the HIV-HCV patients than in the HCV patients. Conversely, the number of liver CD4+ cells was lower in HIV-HCV patients than in HCV patients. The numbers of CD8+ and CD68+ cells were similar in the 2 groups. Finally, liver HCV load, assessed by immunostaining and reverse-transcription polymerase chain reaction, was similar in the 2 groups. We conclude that in the population of HIV-HCV coinfected patients with low-level immunosuppression referred for HCV treatment, fibrosis is worse than in HCV patients and the proportion of CD4+ lymphocytes among CD3+ cells is markedly decreased in the liver, whereas intrahepatic viral load is similar. Our data confirm the need to treat such patients against HCV, and suggest that HIV infection could favor fibrosis via the modulation of the intrahepatic immune response. PMID- 15343511 TI - MCL-1 expression in B-cell non-Hodgkin's lymphomas. AB - B-cell non-Hodgkin's lymphomas are known to express BCL-2 family proteins, of which the myeloid cell leukemia-1 (MCL-1) protein is a member. MCL-1 is involved in viability and immortalization of normal and neoplastic B cells, and expression is regulated transcriptionally and posttranscriptionally, resulting in an anti apoptotic (full length) or a pro-apoptotic (short isoform) gene product. In this study, we assessed 151 B-cell lymphomas for MCL-1 expression and analyzed for expression of the full-length and short isoforms of MCL-1 in B-cell lymphoma cell lines. By using immunohistochemistry, a subset of neoplasms in 9 lymphoma types studied expressed MCL-1, but expression was more frequent and intense in high grade (43 of 49, 88%) compared with low-grade (34 of 92, 37%) lymphomas (P < 0.0001). In follicular lymphomas, MCL-1 expression positively correlated with increasing grade; 1 (14%) of eight grade 1, 7 (70%) of ten grade 2, and all 9 (100%) grade 3 were positive (P < 0.0008). All plasma cell myeloma cases assessed were also MCL-1 positive. By using Western blot analysis, 6 of 7 high-grade B cell lymphoma cell lines showed predominant expression of full-length MCL-1, compared with no or weak expression of the short isoform. One myeloma and 1 of 2 mantle cell lymphoma cell lines also tested showed only full-length isoform expression. Our data suggest that MCL-1 is frequently expressed in high-grade B cell lymphomas and plasma cell myeloma, most likely in its full-length isoform that is an active anti-apoptotic gene product. MCL-1 expression also correlates with grade and may contribute to transformation in follicular lymphomas. PMID- 15343512 TI - The STK11/LKB1 Peutz-Jegher gene is not involved in the pathogenesis of sporadic sex cord-stromal tumors, although loss of heterozygosity at 19p13.3 indicates other gene alteration in these tumors. AB - Germ line mutations in the STK11/LKB1 tumor-suppressor gene (chromosome 19p13.3) are responsible for the Peutz-Jeghers syndrome (PJS). PJS patients frequently develop neoplasms of various organs. Ovarian sex cord-stromal tumor (SCST) with annular tubules, which shows a characteristic morphology intermediate between granulosa cell and Sertoli cell tumors, is distinctively associated with PJS. Although somatic mutations of STK11 are reportedly rare in sporadic forms of common cancers linked to PJS, there are no available studies assessing STK11 alterations in larger series of sporadic ovarian tumors with granulosa, Sertoli or combined differentiation. We examined 29 sporadic SCSTs for loss of heterozygosity (LOH) at 19p13.3, mutation, and promoter methylation of STK11. LOH at 19p13.3 was detected in 12 of 29 (41%) SCSTs, with the highest frequency at chromosome marker D19S894, which localizes approximately 3 mb centromeric to STK11, and it was more frequent in granulosa/Sertoli-stromal cell tumors (10 of 19; 52%) than in thecoma-fibroma tumors (2 of 10; 20%). The 2 fibrothecomas harboring LOH contained sex cord elements. None of the LOH-positive SCSTs demonstrated mutations or promoter methylation of STK11. Our results indicate that LOH at 19p13.3 in sporadic SCSTs targets a gene different from STK11, and may play a role in the pathogenesis of sporadic SCSTs, especially in tumors containing sex cord derivatives. PMID- 15343513 TI - Loss of heterozygosity reveals non-VHL allelic loss in hemangioblastomas at 22q13. AB - Hemangioblastomas (HBs) are low-grade (World Health Organization grade I/IV) central nervous system (CNS) tumors that frequently contain VHL (3p26) mutations. They occur sporadically and in von Hippel Lindau (VHL) disease. Encoded pVHL aids degradation of hypoxia-inducible factors (HIFs) in the presence of normal oxygen levels. HBs provide an in vivo view of HIF effects within a CNS tumor. Typically, HBs are cystic tumors containing a mural nodule formed by noninvasive, vacuolated stromal cells that are embedded in a network of capillaries. Nine HBs, consecutively resected from 8 patients at our institution during a recent 2-year time span, were evaluated for additional losses of tumor suppressor genes. Non VHL microsatellites studied for loss of heterozygosity (LOH) are near tumor suppressor genes lost in gliomas, pituitary adenomas, several CNS tumors on 22q, neurofibromatosis 1, and colon carcinomas (13, 2, 2, 1, and 2 markers for each, respectively). LOH in the region of 3p21.3-3p26.3 occurred in 3 of 8 HBs informative for at least 1 marker (D3S1539, D3S2303, or D3S2373). By using 2 markers (D22S417 and D22S532) for 22q13.2, LOH was found in 5 of 8 informative HBs. All 3 HBs with allelic losses near VHL also showed LOH at 22q13.2. No consistent losses were found with markers for 1p34, LMYC, 5q21, 5q32, 9p21, 10q23, 17p13, and 19q13. LOH for the 22q13.2 region in HBs suggests that the loss of another tumor suppressor gene is involved in the pathogenesis of HBs in addition to VHL. Absence of LOH for glioma markers is consistent with the low grade behavior of HBs. PMID- 15343514 TI - Surgical pathology of infected aneurysms of the descending thoracic and abdominal aorta: clinicopathologic correlations in 29 cases (1976 to 1999). AB - Infected aortic aneurysms are uncommon, and only rarely have their surgical pathological features been described. Clinical and histopathologic features were evaluated in patients undergoing surgical repair of infected aneurysms of the descending thoracic or abdominal aorta over a 24-year period. Findings were compared with observations (primarily from autopsy studies) from the previous 25 year period (1950 to 1975) and other more recent reports. Of the 29 patients in our study, 79% were men, 90% had risk factors for atherosclerosis, and 72% had an identifiable risk or source of infection. Fever was present in 76%, and abdominal or back pain was seen in 66%. Among the 20 cases with an identifiable causative organism, staphylococcus accounted for 30%, streptococcus for 20%, salmonella for 20%, Escherichia coli for 15%, and other organisms for 15%. Aneurysms were saccular in 59% and infrarenal in 42%, and had a mean diameter of 5.6 cm. Microscopically, 6 patterns were recognized: acute inflammation superimposed on severe chronic atherosclerosis (55%), atherosclerosis with chronic inflammation (20%), acute inflammation without atherosclerosis (7%), chronic adventitial inflammation (7%), pseudoaneurysm formation (7%), and necrotizing granulomatous inflammation (4%). Special stains for organisms were positive in only 38% of the cases. Pathological findings of this series of surgical specimens spanning the fourth quarter of the twentieth century were not appreciably different from those described in autopsy series from the preceding years, although the causative microorganisms and agents used to treat them, preoperative diagnostic modalities, and surgical approaches have evolved. PMID- 15343515 TI - The use of morphological characteristics and texture analysis in the identification of tissue composition in prostatic neoplasia. AB - Quantitative examination of prostate histology offers clues in the diagnostic classification of lesions and in the prediction of response to treatment and prognosis. To facilitate the collection of quantitative data, the development of machine vision systems is necessary. This study explored the use of imaging for identifying tissue abnormalities in prostate histology. Medium-power histological scenes were recorded from whole-mount radical prostatectomy sections at x 40 objective magnification and assessed by a pathologist as exhibiting stroma, normal tissue (nonneoplastic epithelial component), or prostatic carcinoma (PCa). A machine vision system was developed that divided the scenes into subregions of 100 x 100 pixels and subjected each to image-processing techniques. Analysis of morphological characteristics allowed the identification of normal tissue. Analysis of image texture demonstrated that Haralick feature 4 was the most suitable for discriminating stroma from PCa. Using these morphological and texture measurements, it was possible to define a classification scheme for each subregion. The machine vision system is designed to integrate these classification rules and generate digital maps of tissue composition from the classification of subregions; 79.3% of subregions were correctly classified. Established classification rates have demonstrated the validity of the methodology on small scenes; a logical extension was to apply the methodology to whole slide images via scanning technology. The machine vision system is capable of classifying these images. The machine vision system developed in this project facilitates the exploration of morphological and texture characteristics in quantifying tissue composition. It also illustrates the potential of quantitative methods to provide highly discriminatory information in the automated identification of prostatic lesions using computer vision. PMID- 15343516 TI - The expression of insulin-like growth factor-I receptor correlates with Fuhrman grading of renal cell carcinomas. AB - Recent reports have shown significant correlation between Fuhrman nuclear grade of renal cell carcinoma (RCC) and patient survival. However, no one specific gene alteration has yet been described to account for this correlation. This study investigated the expression of the insulin-like growth factor-I receptor (IGF-IR) in RCC and correlated the results to the tumor Fuhrman nuclear grade. Formalin fixed, paraffin-embedded sections from 68 cases of RCC were stained using the immunohistochemical avidin-biotin-peroxidase method. An anti-human IGF-IR rabbit polyclonal antibody was used. The stains were semiquantitatively evaluated using the Allred score system, assessing intensity of stain and percentage of positive tumor cells. Statistical analysis was performed using the Kruskal-Wallis test. Strong and diffuse cytoplasmic IGF-IR stain (Allred score 7 to 8) was identified in 25 of 25 (100%) of grade 3 and 4 RCCs. Grade 2 RCCs had a median IGF-IR Allred score of 4. Ten of 10 (100%) grade 1 RCCs were negative. Even in the positive high-nuclear-grade tumors, areas of low nuclear grade, when present, were IGF-IR negative. Statistical analysis using the Kruskal-Wallis test demonstrated significant correlation between increasing Fuhrman nuclear grade and increasing IGF-IR Allred score (P <0.0001). Thus we report the novel finding of significant statistical correlation between IGF-IR protein expression and Fuhrman nuclear grade of RCC, and consequentially with patient survival. PMID- 15343517 TI - Silent corticotroph adenomas: further clinical and pathological observations. AB - Adrenocorticotroph cell pituitary adenomas immunoreactive for adrenocorticotropic hormone (ACTH) but unassociated with preoperative signs of hypercortisolism constitute between 6% and 43% of all ACTH adenomas. Few large series have been published. At our referral center for pituitary diseases, we have encountered 12 patients with silent ACTH adenomas, none of whom exhibited definite clinical features of hypercortisolism preoperatively. Two patients presented with apoplexy, and in 2 patients preoperative neuroimaging studies mimicked craniopharyngioma. Pathological examination revealed 8 adenomas with variably basophilic cytoplasm (type I, including 1 each with coarse basophilic granules and Crooke's hyaline change) and 4 with predominantly chromophobic cytoplasm (type II). Diffuse versus patchy (30% to 50% of cells) immunostaining best distinguished these 2 types; calcitonin staining was focal or negative in both. Two patients had unexpected postoperative courses consistent with acute cortisol insufficiency; 1 patient suffered from a severe flu-like illness, and the other had dizziness and was found to have a serum cortisol level of < 1.0 microg/dL. Both patients improved after cortisol replacement followed by a slow taper. Another patient developed 2 separate pituitary adenomas, a silent ACTH adenoma followed by a pure prolactinoma resected months later. Clonality studies demonstrated that the 2 tumors had arisen from different clonal populations. These cases offer additional insights into clinical, neuroimaging, histological, and biological features of silent ACTH adenomas. Because 2 of these patients seemed to require postoperative cortisol supplementation that otherwise would not have been given, clinicians should be notified about ACTH immunostaining in adenomas from patients without preoperative diagnoses of Cushing's disease, to optimize postoperative care. PMID- 15343518 TI - E-cadherin-beta-catenin adhesion complex in neuroendocrine tumors of the lung: a suggested role upon local invasion and metastasis. AB - Dysfunction or loss of the intercellular adhesion complex E-cadherin-beta-catenin is frequent in non-small cell lung carcinomas in which E-cadherin and beta catenin loss has been considered to be a molecular marker of tumor progression and poor prognosis. With an aim of evaluating the expression of the E-cadherin beta-catenin complex and its prognostic role in neuroendocrine tumors (NET) of the lung, immunohistochemical analysis was performed in 102 NET, including 16 low grade typical carcinoids, 8 intermediate-grade atypical carcinoids, 37 large-cell neuroendocrine carcinomas (LCNEC), and 41 small-cell lung carcinomas, both high grade tumors. Impaired E-cadherin expression (loss or cytoplasmic delocalization) was observed in 80 (78%) of 102 samples, and impaired beta-catenin expression was noted in 74 (72%) of 102 cases. The impaired expression of E-cadherin and beta catenin was observed with a higher frequency in high-grade tumors (87% and 83%, respectively) than in carcinoids (50% and 37%, respectively; P < 0.0001). Impaired expression of the E-cadherin and beta-catenin molecules also correlated with lymph node metastasis (P = 0.0001 and P = 0.0005, respectively) and with advanced stage disease (P < 0.0001 for both factors). Moreover, impaired E cadherin expression directly correlated with an extensive disease in carcinoids and in LCNEC (P = 0.02 and P = 0.04, respectively) and with node metastasis in LCNEC (P = 0.01). Levels of E-cadherin and beta-catenin were correlated with each other, consistent with an internal regulatory loop. Our results indicate that down-regulation of the E-cadherin-beta-catenin complex plays a role in NET progression. PMID- 15343519 TI - Atypical extraventricular neurocytoma with oligodendroglioma-like spread and an unusual pattern of chromosome 1p and 19q loss. AB - An insular cortex tumor in a 54-year-old woman showed unequivocal neurocytic features, including open nuclei, distinct nucleoli, and strong synaptophysin immunoreactivity. Ultrastructurally, there were neuritic-type processes with microtubules and hillock-like attachments, and there were dense-core granules. Atypical features were mitotic activity, prominent vasculature, and small foci of necrosis. Peripherally, there was oligodendroglia-like histology with single-cell infiltration of white matter and perineuronal spread in cortex. Fluorescence in situ hybridization analysis with chromosome 1 and 19 probes showed 3 copies of 1q and 2 copies of 1p and showed 2 copies of 19q and 4 copies of 19p. This yielded a 1p-19q loss of heterozygosity pattern similar to that seen in oligodendrogliomas, although the actual chromosomal abnormality is distinct. This tumor, best classified as an atypical neurocytoma with oligodendroglia-like spread, supports suggestions of a close histogenic relationship between oligodendroglial and neurocytic tumors. This case also illustrates the limitations of relying exclusively on loss of heterozygosity analysis for tumor classification. PMID- 15343520 TI - Eosinophilic myocarditis in a patient with idiopathic hypereosinophilic syndrome: insights into mechanisms of myocardial cell death. AB - Idiopathic hypereosinophilic syndrome (HES) consists of a prolonged state of eosinophilia of unknown origin with organ involvement. We describe the case of a patient who developed fatal eosinophilic myocarditis. A 23-year-old woman with an 8-month history of eosinophilia presented with symptoms of myocarditis. Histological evaluation of an endomyocardial biopsy specimen revealed marked endomyocardial eosinophilic infiltration with eosinophil-rich granulomas and areas of myocyte necrosis. A terminal deoxynucleotidil transferase assay revealed apoptosis in several cardiomyocytes and vascular cells, mainly in the myocardial areas with higher eosinophil density. Evaluation of an endomyocardial biopsy specimen obtained after steroid therapy demonstrated that the eosinophils had disappeared, but there was marked myocardiosclerosis and scattered apoptotic cells. The patient slowly developed heart failure and died of sudden arrhythmic death. HES can cause severe myocarditis with extensive myocyte loss, probably due to both necrosis and apoptosis. Myocardial fibrosis may occur despite treatment, and patients may be at risk for fatal arrhythmias. PMID- 15343521 TI - A case of primary mucosa-associated lymphoid tissue lymphoma of the vagina. AB - We report the first case of primary mucosa-associated lymphoid tissue (MALT) lymphoma of the vagina, the diagnosis of which is supported by genetic and immunophenotypic studies. A 65-year-old, para 2 woman presented to our hospital in July 1997 with a history of prolonged vaginal discharge. Although cytologic examination suggested possible malignancy, a biopsy of the vaginal wall was diagnosed as chronic inflammation. In June 2000, she underwent gynecologic examination because of anuria. Excisional biopsy revealed subepithelial infiltration of atypical lymphoid cells that stained for CD20, CD79a, and BCL-2; stained weakly for IgM; and did not stain for CD3, CD5, CD7, CD10, CD56, CD23, and IgD, suggesting marginal zone B-cell lineage. Monoclonality was detected by Southern blot analysis, and this patient was finally diagnosed as having primary MALT lymphoma of the vagina. She received 3 cycles of chemotherapy (THP-COP) and concurrent radiation to the whole pelvis. The patient is alive and well 40 months after treatment. Because the vagina is one of the mucosa-associated tissues, MALT lymphoma, though rare, must be included in the differential diagnosis of the vaginal neoplasms. PMID- 15343522 TI - Prognostic factors in Bilharzial-related bladder cancer. PMID- 15343524 TI - Fifteen-year clinical survivorship of Harris-Galante total hip arthroplasty. AB - We studied the long-term outcome of uncemented total hip arthroplasty using HG-I components in 90 hips (80 patients) with an average age of 57.5 years who were operated on between 1984 and 1986 at our institution. Average follow-up was 14.9 years, and no patient was lost to follow-up. Hip scores improved significantly, and there was clinical and radiographic evidence of bony ingrowth on the acetabular components in all patients. There were 11 revisions in the study population for reasons related to failure of the femoral component (8 hips), acetabular liner dissociation (2 hips), and deep infection (1 hip). At the latest follow-up, 3 femoral components were determined to be loose. No acetabular component was revised for aseptic loosening. In addition to the revisions, there were 2 reoperations, 1 for psoas tendon release, and 1 for excision of heterotopic ossification. Thus, the survivorship free of revision and free of mechanical failure for the acetabular component at 15 years was 95.7% (95% confidence interval [CI], 0.89-1.0) and 91.9% (95% CI, 0.83-0.98), respectively. The survivorship at 15 years for the femoral component was 86.8% (95% CI, 0.78 0.95) free of revision and 82.0% (95% CI, 0.71-0.92) free of mechanical failure. PMID- 15343525 TI - Mobile-bearing total knee prosthesis: a 5- to 9-year follow-up of the first 110 consecutive arthroplasties. AB - This study regards the total articulating cementless knee. This is a mobile bearing knee, the tibial component of which consists of 2 parts: a highly conforming polyethylene insert freely rotating on a metal tray. Our case study relating to the implant of the first 110 knees operated on consecutively from 1991 to 1995 is reported, with an average follow-up of 6.3 years (range, 5-9 years). The average preoperative Knee Society Score was 78 points, and the average postoperative score was 156 points. The complications specifically related to the prosthetic components and which required revision surgery were 4: 2 cases of instability, 1 aseptic loosening of the tibial tray, and 1 traumatic dislocation of the tibial insert. A further 3 patients underwent reoperation for causes not strictly related to the implant: 2 because of intractable patellar pain and 1 because of periprosthetic ossifications that limited flexion. All of the complications were observed in patients operated on during the first 3 years of our experience, thus suggesting a definite learning curve with this prosthesis. No evidence of progressive radiographic periprosthetic osteolysis was recorded, and no relevant polyethylene wear was observed over time. Kaplan-Meier survival curves show the probability of survival to be 93.7% with revision surgery for any reason as an endpoint, and 96.3% with revision surgery for a mechanical reason as an endpoint. Certainty that mobile-bearing total knees are able to assure a longer life of the implant than the conventional models would require an evaluation of results over 15 to 20 years. However, in the meantime, these good preliminary results at least justify continuing the use of this type of prosthesis, which still awaits confirmation of the, as-yet-theoretical, advantage compared with fixed-bearing total knees. PMID- 15343526 TI - Early instability with mobile-bearing total knee arthroplasty: a series of 25 cases. AB - Between December 1987 and January 2002, 25 cases of clinical instability following mobile-bearing total knee arthroplasty (TKA) with meniscal bearings or rotating platforms presented for evaluation at our institution. These cases were retrospectively identified. All were performed at outside institutions by a variety of surgeons. All clinical examinations were performed by the authors. Nine cases were revised at our institution. All 25 cases had clinical evidence of severe coronal-plane instability and pain. Eight cases had polyethylene dislocation or subluxation that was evident radiographically and clinically. Four cases had extensor-mechanism dysfunction. Eighteen cases had symptoms immediately postoperatively. Twenty-three of the 25 cases had symptoms within 2 years' postoperatively. Any potential long-term benefit of design innovations must be balanced with known problems leading to early failure. PMID- 15343527 TI - Relationship between surgical volume and early outcomes of total hip arthroplasty: do results continue to get better? AB - This retrospective study explored the relationship between the volume of total hip arthroplasties (THA) and postoperative mortality and early complications from a single institution. One thousand hip arthroplasties in 932 patients were identified during a 1-year period, which included 786 primary and 214 revision hip arthroplasties. The postoperative 6-month combined medical and orthopedic complication rate for primary and revision THA was 7.9% and 16.5%, respectively. The 6-month mortality rate for the overall group was 0.5% (5 deaths), for the primary hips was 0.4% (3 deaths), and for the revision hip was 0.9% (2 deaths). The mortality and complication rates of many surgical procedures, including joint arthroplasties, are inversely related to hospital and surgical volume. The reduction in complication rate, however, approaches a plateau and does not improve regardless of an increase in the surgical and hospital volume. PMID- 15343528 TI - Comorbid illness affects hospital costs related to hip arthroplasty: quantification of health status and implications for fair reimbursement and surgeon comparisons. AB - Optimized resource allocation, reimbursement negotiations, and provider comparisons hinge on an understanding of the drivers of healthcare costs. Indices of comorbid illness may be useful for stratifying patients based on cost. Total hospital cost was analyzed for 1 surgeon's hip arthroplasty patients (June 1998 March 2001). Three scales of health status were selected as independent predictors. One thousand ninety-two hip arthroplasty inpatient stays were evaluated. The median total hospital cost was 14,011 dollars. An increasing burden of comorbid illness as measured by the All Patient Refined Diagnosis Related Group Severity of Illness scale and the modified Charlson Comorbidity Index was significantly associated with increasing hospital cost. Comorbid illness is associated with cost; scales may be used to stratify patients based on risk of high cost care. PMID- 15343529 TI - Bulk femoral-head autografting in uncemented total hip arthroplasty for acetabular dysplasia: results at 8 to 11 years follow-up. AB - Acetabular dysplasia causes difficulty in achieving adequate coverage of the acetabular component during total hip arthroplasty (THA). Bulk femoral-head autografting is one technique that has been used to achieve better coverage of the acetabular component. Long-term follow-up studies have shown a significant failure rate when this technique has been used in conjunction with a cemented acetabular component; however, with uncemented components, early results have been encouraging. In our study, 15 patients with acetabular dysplasia underwent uncemented THA, during which bulk femoral-head autografts were used. At an average follow-up of 10 years, no cases required revision, and radiologically, the bone graft had united. Our results support the use of bulk femoral-head autografting in patients with acetabular dysplasia requiring hip arthroplasty. PMID- 15343530 TI - Radiographic changes in the patella following quadriceps turndown for revision total knee arthroplasty. AB - Quadriceps turndown is a technique that may be used to enhance exposure of the tight total knee arthroplasty (TKA), particularly in the revision scenario. This technique does, however, compromise the vascularity of the patella, with avascular necrosis (AVN) being a possible sequela. A modified Coonse-Adams quadriceps turndown was performed in 29 revision TKAs in 27 patients. Immediate preoperative and sequential postoperative radiographs were analyzed for changes in the patella, including sclerosis, flattening, fracture, and fragmentation, as evidence of possible AVN, and clinical scores were collected prospectively. Eight patellae had such radiographic changes. Despite a lack of corresponding worsening in clinical outcome in these patients, quadriceps turndown is shown to have a high risk of subsequent changes consistent with patellar AVN, and should be avoided when other available techniques can achieve satisfactory exposure. PMID- 15343531 TI - Femoral nerve block for total knee arthroplasty patients: a method to control postoperative pain. AB - This study was designed to determine the effects of a single-injection femoral nerve block (FNB) using 30 mL of 0.5% bupivacaine with epinephrine 1:200,000, on pain control following total knee arthroplasty (TKA). Forty patients were randomly distributed into 2 groups: Group A received general anesthesia plus a FNB (n = 19), whereas Group B received general anesthesia plus a FNB with 30 mL of preservative-free saline (n = 21). The amount of morphine used, sedation, and average pain perception were measured for the first 24 hours and daily postoperatively. Group A used significantly less morphine (48.1 mg) compared with Group B, which used 76.2 mg during the first 24 hours after surgery (P = 0.003). Group A's sedation scale was significantly less than group B's (2.26 vs 2.67) (P = 0.045). The average pain perception was significantly different (P =.002). Postoperative management of pain following TKA can be improved through a preoperative single-injection FNB with 0.5% bupivacaine plus epinephrine 1:200,000. The cost is minimal, risks appear acceptable, and the procedure is efficacious. PMID- 15343532 TI - Cemented revision hip arthroplasty using strut and impacted cancellous allografts. AB - The purpose of this study was to evaluate the hypothesis that the mechanical failure rate is reduced by the combined use of cortical strut and impacted cancellous allografts. There were 10 men and 14 women in the series, with a mean age at the time of the index revision of 56.6 years (range, 43-69 years). They were followed up for a minimum of 5 years (mean, 5.8 years; range, 5-7 years). The Harris Hip Score improved from a mean of 26 points (range, 10-37 points) before revision surgery to a mean of 83 points (range, 45-100 points) at the latest review. The mean visual analogue scale to pain was 8.4 (4-10), and the mean satisfaction visual analogue scale was 8.7 (4-10). WOMAC scores at the latest review correspond to mild pain, mild stiffness, and only moderate difficulty with physical activity. One femoral stem (4%) had aseptic loosening, and it was revised. Two hips (8%) had infection. All allografts were incorporated to the host femur. The combination of proximal onlay strut grafts with impaction bone grafting in femora with proximal segmental bone defects is associated with very acceptable results at midterm follow-up. PMID- 15343533 TI - Efficacy of abduction bracing in the management of total hip arthroplasty dislocation. AB - A retrospective review of total hip arthroplasty (THA) dislocations was performed to determine the effectiveness of abduction bracing following closed reduction. Patients were grouped as a first-time dislocation (n = 91) or recurrent dislocation (n = 58) and whether or not they received an abduction brace; re dislocation defined failure of treatment. The mean follow-up was 4.0 years in the first-time group and 3.7 years in the recurrent group. Among patients treated for first-time dislocations, 61% re-dislocated with a brace and 64% of nonbraced patients re-dislocated. In the recurrent group, 55% re-dislocated with a brace, whereas 56% re-dislocated without a brace. Chi-square analysis revealed that observed differences were not significant. There was no significant difference among groups with regards to age, sex, operative side, or significant surgical parameters. Abduction bracing following closed reduction of THA dislocation is ineffective in preventing re-dislocation. PMID- 15343534 TI - Insertion of an expandable cement restrictor reduces intramedullary fat displacement. AB - The insertion of an intramedullary cement restrictor can lead to the same pathophysiologic reactions that are well described as fat embolism syndrome after cement and stem implantation. Fat and marrow are forced into the draining veins of the femur. In this study, we compared the extrusion of intramedullary contents produced by the insertion of a commonly used gelatin (Biostop G/IMSET) and a new expandable cement restrictor (REX Cement Stop) in 7 paired, fresh-frozen femora. A further 9 pairs were used to evaluate the function of a custom-made application instrument, which allowed for suction during plug (Biostop G/IMSET) insertion. The oversized restrictor produced, on average, twice as much fat as the expandable restrictor (P = 0.0156). The gelatin plug with additional suction was not associated with reduced fat extrusion, because the suction device in its current design failed because of obstruction in most cases. The expandable restrictor showed favorable characteristics and has the potential to reduce the risk of fat embolism. PMID- 15343535 TI - Bone viability determination in human cancellous bone from patients undergoing revision hip arthroplasty. AB - In December 2000, Sulzer Orthopedics Inc. announced the recall of specific lots of Inter-Op acetabular components as a result of an adverse tissue response to a contaminant on the porous surface of the component. This study was initiated to assess bone viability surrounding the primary and revision components. Twelve patients were placed on a double-tetracycline labeling protocol before revision surgery. Initial and final acetabular reamings were frozen-sectioned using cryostat technology. A mineral apposition rate, preoperative and postoperative radiographs, and Harris Hip Scores were evaluated. The tetracycline data showed that bone viability was not compromised at initial or final reaming levels in this series of patients. Significant improvement was observed in postoperative Harris Hip Scores when compared with preoperative Harris Hip Scores. PMID- 15343536 TI - Results of revision total knee arthroplasty after exposure of the knee with extensor mechanism tenolysis. AB - The purpose of this research was to investigate the utility of 1 surgical exposure method, extensor mechanism tenolysis (EMT), for total knee revision, and to evaluate complications associated with this technique. Two hundred seven knee revisions were performed on 198 patients (9 bilateral) over a 3-year period (September 1997 to October 2000). The surgical exposures used were EMT in 203 cases, V-Y quadricepsplasty in 2 knees, and patellectomy in 2 additional cases. The complications associated with EMT were peripatellar fibrosis requiring arthroscopy, 7.2%; hematoma, 4.8%; stiffness requiring manipulation, 3.9%; patellar subluxation, 1.4%; extensor lag >5 degrees, 1.0%; quadriceps tendon rupture, 0.5%; and instability, 0.5%. EMT is associated with a low complication rate comparable with or better than other exposure methods. PMID- 15343537 TI - "Wandering resident" surgical exposure for 1- or 2-stage revision arthroplasty in stiff aseptic and septic knee arthroplasty. AB - A retrospective study was performed on 18 patients who had undergone revision total knee arthroplasty (TKA) between 1992 and 2000 with stiff knee (average, 50 degrees range of motion). The so-called "wandering resident" surgical exposure was used. This procedure was used twice in 5 cases with low-grade infection in a 2-stage revision of their septic prosthesis. Follow-up was from 1 to 8 years (average, 3.5 years). All patients had good clinical results, with an 86 degrees average range of motion. The Knee Society Score preoperatively ranged from 35 to 52 (average, 40) and improved to 72 to 89 (average, 84). Postoperative rehabilitation was slower, requiring the use of a knee brace in extension for 6 weeks for support while the patient ambulated and without this support for daily physiotherapy and continuous passive motion at 0 degrees to 70 degrees. Full range of motion with no brace was started after 6 weeks. PMID- 15343538 TI - Spacer endoprosthesis for the treatment of infected total hip arthroplasty. AB - We reviewed the treatment of infected total hip arthroplasty with a temporary spacer endoprosthesis. To fabricate the spacer, antibiotic-loaded cement was inserted into a specially designed mold. A central rod pin was superficially imbedded as an endoskeleton once the cement reached a doughy state. After polymerization, the final product was removed from the mold and inserted as an articulating spacer. Twenty patients were followed for an average of 38 months (range, 26-67 months). There were no recurrent or persistent infections. Eighteen patients underwent a successful 2-stage exchange. Two patients retained the spacer as a definitive treatment method. Complications with the spacer included 2 fractures and 2 dislocations. Overall, this cost-effective technique provided efficient local antibiotic delivery, early mobilization, facilitation of reimplantation, and improved patient satisfaction. PMID- 15343539 TI - Antibiotic-loaded articulating cement spacer in the 2-stage exchange of infected total knee arthroplasty. AB - An antibiotic-loaded articulating cement spacer (ALACS) was used in the 2-stage exchange of infected total knee arthroplasty. Specially designed molds produced articulating femoral and tibial spacer components. Twenty-four consecutive patients were followed for an average of 33 months (range, 28-51 months). Two patients (8%) had a persistent infection after the first stage. Twenty-two patients (92%) underwent a successful 2-stage exchange. Minimal soft-tissue contracture and minimal bone loss were encountered during reimplantation. None of these patients developed a recurrent or persistent infection. The average postoperative knee flexion was 104 degrees (range, 89 degrees-122 degrees). The average Hospital for Special Surgery score was 82 (range, 63-96). The ALACS spacer preserved knee function between stages, resulting in effective treatment of infection, facilitation of reimplantation, and improved patient satisfaction. PMID- 15343540 TI - The stability of the cemented tibial component of total knee arthroplasty: posterior cruciate-retaining versus posterior-stabilized design. AB - Micromotion of the tibial component in 40 knee arthroplasties for gonarthrosis was studied using Roentgen stereophotogrammetric analysis. The stability of this component was assessed for 2 years' postoperatively. In all arthroplasties, an attempt was made to reconstruct the preoperative posterior slope. Posterior cruciate-retaining (CR) and posterior-stabilized (PS) components showed at 2 years a maximum total point motion of 0.6 +/- 0.4 mm and 0.7 +/- 0.5 mm, respectively. Whereas 92.5% of the implants were determined to be stable, 1 of the CR group and 2 of the PS group displayed migration between the first and the second year of at least 0.2 mm. A negative correlation between subsidence of the tibial component at 2 years of follow-up and the difference between preoperative and postoperative tibial slope was found. Consequently, we suggest that restoring the original posterior slope of the tibial plateau must be a goal of tibial component implantation. PMID- 15343541 TI - Progression of radiolucent lines in cementless twin-bearing low-contact-stress knee prostheses: a retrospective study. AB - A total of 134 low-contact-stress (LCS) total knee arthroplasties (TKAs) with mobile meniscal bearings were implanted in 121 patients with a mean age of 74 years (range, 49-91 years) at the time of surgery. The radiographic results obtained from 91 prostheses at an average follow-up of 7.5 years (range, 4.8-9.6 years) showed that radiolucent lines (RLL) appeared most frequently near the tibial plateau (97%) and that most RLL were present immediately postoperatively (67%) or appeared within the first year (96%). RLL were predominantly nonprogressive (99%). Progressive RLL did not affect the fixation or stability of the prosthesis. There were no revisions for aseptic loosening. Cementless LCS TKA achieves excellent radiologic and clinical midterm results. PMID- 15343542 TI - Late sciatic nerve palsy caused by hematoma after primary total hip arthroplasty. AB - A case of late sciatic nerve palsy caused by subfascial hematoma after uncemented right total hip arthroplasty is reported. The patient developed respiratory distress 13 days postoperatively and was admitted to another institution, where she was diagnosed with pulmonary embolism and was subsequently therapeutically anticoagulated with heparin. The patient complained of right-leg numbness and tingling 18 days' postoperatively, which progressed to complete sciatic nerve palsy over several hours. PMID- 15343543 TI - Advanced osteoarthritic knee with neglected patellar tendon rupture treated with total patellectomy and total knee arthroplasty. AB - Loss of the knee extensor mechanism results in a change of normal knee joint alignment and functional anteroposterior instability. In patients with neglected or chronic patellar tendon rupture, advanced degenerative change of the knee joints may develop at the later stage. We present a case of a 64-year-old man with chronic left patellar tendon rupture and 10-cm proximal patella migration associated with advanced osteoarthritis of the knee. Total patellectomy and simultaneous total knee arthroplasty (TKA) relieved his symptoms and disability successfully. His left knee still did well at 7-year follow-up. PMID- 15343544 TI - Early tibial tray failure of a Duracon knee with retrieval analysis. AB - We report a case of early tibial tray fracture of a Duracon knee prosthesis. Aside from the clinical, radiographic, and gross analysis of the failed prosthesis, we also performed analysis on the retrieved polyethylene component and the fractured tibial baseplate. In the analyses, we noted significant femoral component malalignment, uneven tray cementation, and inherent metallurgical weakness. It appears that the high compressive load on the medial tray resulted in bending fatigue failure. To avoid this complication, it is important to restore the normal alignment of the knee joint and use a polyethylene insert of higher conformity, at least 6 mm thick. PMID- 15343545 TI - Emergency hemipelvectomy as a result of uncontrolled infection after total hip arthroplasty: two case reports. AB - Described are 2 patients with sepsis caused by an infection of a total hip arthroplasty. In both cases, the infection involved spread to the ipsilateral retroperitonial and proximal femur soft tissues, and substantial parts of the osseous hemipelvis and proximal femur. Furthermore, the infection could not be controlled despite repeated surgical interventions. Progression of the infection of soft tissue and bone resulted in severe bone stock loss, contractures of hip and knee, and severe neurologic deficit of the involved limb. Hemipelvectomy was indicated as a life-saving procedure. Reluctance to perform vigorous debridement, including removal of the components, can lead to a local spread of the infection, resulting in an uncontrollable process leading to a life-threatening situation. PMID- 15343546 TI - The clinician and the researcher. PMID- 15343547 TI - Intervening as a passenger in drinking/driving situations. AB - This study sought to query adolescents about drinking/driving situations and interventions used in these circumstances. A human science qualitative method was used to analyze descriptions of situations and interventions to gain the perspective of the life world of the young person. Findings include the drinking/driving situations of entangled, endangered, and stranded. Passenger interventions included persuading, interfering, planning ahead, and threatening. Practice, education, and research implications for those who work with adolescents are offered. PMID- 15343548 TI - Heart failure patient learning needs after hospital discharge. AB - This descriptive-correlational study examined the perceived learning needs of heart failure patients in postdischarge settings, as well as the influence of demographic variables on these learning needs. The Outpatient Heart Failure Learning Needs Inventory was used to rate subjects' perceptions of the importance of educational topics on a five-point Likert scale. Findings indicated that subjects perceived signs and symptoms and medications as most important to learn and diet, activity, and psychological factors as least important to learn. These findings are consistent with previous research and provide a framework on which to base the development of educational programs for patients with heart failure. A significant finding was that nearly 25% of screened patients were unable to participate because they were unaware that they had been diagnosed with heart failure. PMID- 15343549 TI - Hypertension and diabetes in Detroit Hispanics. AB - The purpose of this study was to explore the prevalence of hypertension and diabetes alone and together in a sample of Hispanics. There were 111 participants with a mean age of 51 years. Twenty-six percent of the participants were hypertensive, a proportion well above the Healthy People 2010 target of 16%. The majority of hypertensive participants had isolated systolic hypertension, which is consistently associated with greater cardiovascular risk. Thirty-seven percent of the hypertensives were also diabetic. Diabetics were significantly more likely than non-diabetics to have stage 3 hypertension than either stage 1 or stage 2 (chi(2) [1] = 7.17, p <.01). It is important for nurses to screen Hispanic clients who are 18 years and older for high blood pressure. Early case finding will help nurses increase awareness and control of high blood pressure among Hispanics, which is crucial to avoid the enormous human and financial burdens of cardiovascular morbidity and mortality. PMID- 15343550 TI - Adolescent scoliosis: effects of corrective surgery, cognitive-behavioral interventions, and age on activity outcomes. AB - The effects of spinal fusion surgery and cognitive-behavioral interventions on 88 adolescents' (11-18 years) activity outcomes were examined using a randomized trial with three intervention groups (information only, coping only, coping plus information) and a control group. The effects included a significant drop from baseline (preoperative) in usual activities and social activities at 1 month postsurgery for all groups, indicating that initially postsurgical recovery is particularly disruptive to patients' lives. At 3 months after surgery, all groups showed increased usual activities and social activities. Between the 3- and 6 month assessments, all groups had increases in social activities. Furthermore, the information only group had a significant increase in usual activities from 3 to 6 months. Younger adolescents (ages 11-14) in the combined information plus coping group and the control group had higher social scores over the postsurgery recovery period compared with those in the coping-only group. No differences were found on academic performance. The adolescents did not return to their baseline social activity levels during the 9-month recovery period (typically when long term recovery is completed), indicating that the surgery itself has a long-term negative effect on patients' social life. PMID- 15343551 TI - Health care providers' attitudes toward current food safety recommendations for pregnant women. AB - Semistructured interviews were conducted with 23 health care professionals who work with pregnant women in a preliminary study to understand health care providers' attitudes regarding current food safety recommendations for pregnant women and interest in education and patient materials on food safety during pregnancy. Only 8 of 23 interviewed currently provided food safety information to their pregnant clients. Limited understanding of food safety issues and limited time with patients were barriers to providing such information. Based on their contact time with patients, background, and interest in food safety issues, nurses, visiting nurses, and Special Supplemental Nutrition Program for Women, Infants and Children professionals should be targeted as conveyers of food safety information to pregnant women. PMID- 15343552 TI - CVD prevention strategies with urban and rural African American women. AB - Using Pender's Health Promotion Model, this study tested a work site cardiovascular disease (CVD) risk factor reduction intervention among low-income African American (LAAW) women. Individual CVD risk profiles were identified at the work site through (1) health risk appraisal, (2) blood pressure measurement, (3) body mass index calculation, (4) individual interviews about diet and exercise behaviors, and (5) total cholesterol analysis. Two LAAW groups, one urban and one rural, exhibited higher or similar pretest CVD relative risks (RR) when statistically compared with a national sample of African American women. The LAAW study samples were younger and more educated. Pretest cholesterol and fat intake for the rural women were higher than for the urban women (<.05). Posttest changes in cholesterol and fat intake risks were more significant in rural LAAW than in urban LAAW (<.05). PMID- 15343553 TI - Considerations for preparing collaborative international research: a Ugandan experience. AB - This article describes issues to consider when planning and conducting international research projects. Key considerations include building collaboration, developing a comprehensive and feasible research plan, funding and budgets, addressing human subjects concerns, and analyzing and disseminating project findings. These considerations and related methodological issues are discussed in the context of a replication pilot project conducted outside Kampala, Uganda. Ongoing dialog, flexibility, and collaboration, in addition to good science, are critical to developing successful international research projects. PMID- 15343554 TI - The effect of neuromuscular electrical stimulation on arthritis knee pain in older adults with osteoarthritis of the knee. AB - The objective of this study was to examine the short- and long-term effects of a home-based, 12-week neuromuscular electrical stimulation (NMES) of the quadriceps femoris to decrease arthritis knee pain in older adults with osteoarthritis of the knee. The study sample (N = 38) was randomly assigned to the NMES treatment plus education group or the arthritis education-only group. Pain was measured in both groups with the McGill Pain Questionnaire (MPQ) at baseline, during the intervention at weeks 4, 8, 12, and at follow-up and with the Arthritis Impact Measurement Scale 2-Pain Subscale (AIMS2-PS) at baseline and week 12. The NMES Pain Diary (PD) was completed 15 minutes before and after each stimulation session. There was a significant 22% decline in pain 15 minutes after as compared with immediately before each NMES treatment (p <.001), as measured by the PD. No significant group differences were found between the 2 groups over the course of the intervention and follow-up. These findings indicate that a home-based NMES intervention reduced arthritis knee pain 15 minutes after a NMES treatment. PMID- 15343555 TI - Using focus groups to identify asthma care and education issues for elderly urban dwelling minority individuals. AB - A growing number of older adults are afflicted with asthma; these older asthmatic individuals suffer more deleterious consequences as compared with younger asthmatic individuals. Asthma is a chronic condition requiring the person's ability to self-manage symptoms. Few educational programs have focused on older asthmatic adults' learning needs and even fewer on those of older minority individuals with this disease. Three focus groups were conducted in East Harlem. Each group consisted of 6 to 10 participants. All focus groups were led by the same co-leaders and were conducted using the same semistructured format. Discussions were audio-taped and transcribed. Transcriptions were reviewed by two independent raters who determined major themes and concerns. Comparisons were made and discrepancies resolved through discussion and consensus with the team of investigators. In this pilot study, there were important similarities in the themes identified by participants in the 3 focus groups. Older individuals with asthma, their unlicensed caregivers, and health professionals all identified the following as important in the care of older adults who are asthmatic: (1) the negative impact of asthma on the individual's quality of life, (2) high cost of medications, (3) nonadherence to the medical regimen, and (4) difficulty that these individuals have in accessing the health care system. PMID- 15343556 TI - Transcultural change: a challenge for the public health system. PMID- 15343557 TI - IOM data standards for patient safety and nursing. PMID- 15343559 TI - Carotid stenting with a new system for distal embolic protection and stenting in high-risk patients: the carotid revascularization with ev3 arterial technology evolution (CREATE) feasibility trial. AB - The purpose of this study was to evaluate the feasibility of carotid artery revascularization using a new system for carotid stenting and distal embolic protection in 30 patients with severe carotid stenosis and high risk for carotid endarterectomy (Carotid Revascularization With ev3 Arterial Technology Evolution, or CREATE). Previous studies suggest that patients with carotid stenosis and serious comorbid cardiopulmonary and anatomic conditions are at high risk for carotid endarterectomy. All patients underwent percutaneous revascularization using the Protege GPS self-expanding nitinol stent (ev3, Plymouth, MN) and the Spider distal embolic protection system (ev3). In-hospital and 30-day outcomes were analyzed. High-risk features included age > 75 years (63%), left ventricular ejection fraction < 35% (20%), and restenosis after prior carotid endarterectomy (53%). Procedural success was 100%. In-hospital complications included severe vasovagal reactions in six patients (20%) and a popliteal embolus in one patient (3.3%), treated by successful embolectomy. During 30 days of follow-up, two patients (6.6%) experienced minor neurological deficits, including transient expressive aphasia that resolved without therapy in one patient and homonymous hemianopsia due to contralateral posterior circulation stroke in one patient. This study supports the feasibility of percutaneous carotid artery revascularization with the Protege GPS self-expanding stent and Spider distal embolic protection system, which will be evaluated in a large multicenter pivotal trial (CREATE Pivotal Trial). PMID- 15343560 TI - Wireless laser-assisted angioplasty of the superficial femoral artery in patients with critical limb ischemia who have failed conventional percutaneous revascularization. AB - Percutaneous revascularization has become an effective treatment for patients suffering from chronic critical limb ischemia (CLI) due to chronic atherosclerotic obstructions, including total occlusions. Unlike other vascular beds, total chronic occlusions of the femoropopliteal arteries are frequently found in patients with severe claudication or CLI. As a consequence, patients with long chronic total occlusions of the femoropopliteal arteries are generally not considered optimal candidates for percutaneous revascularization and are frequently referred for surgical revascularization. In the present study, we sought to evaluate the feasibility, safety, and outcome of a modified wireless laser ablation technique to recanalize total occlusions in patients with CLI who had failed conventional percutaneous techniques for limb salvage. Procedural success, complications, actuarial freedom of limb loss, and surgical revascularization were evaluated in 25 patients after a mean follow-up of 13 +/- 8 months. Procedural success was achieved in 21 patients (84%). Actuarial freedom from surgical revascularization or limb loss was 72%. There was one vascular perforation. No deaths or distal embolization occurred. Three patients (12%) required limb amputation during follow-up, whereas four patients (16%) had surgical revascularization in the presence of feasible vascular targets. Limb salvage was achieved in 88% of patients when laser recanalization was combined with surgical revascularization. These results suggest that the use of laser ablation is safe and facilitates angioplasty and stenting in patients with CLI that failed conventional endovascular revascularization. This technique might prevent limb loss in patients with CLI due to femoropopliteal total occlusions, particularly in patients with unsuitable anatomy for surgical revascularization. PMID- 15343561 TI - Rescue excimer laser angioplasty for treatment of critical limb ischemia. PMID- 15343562 TI - Endovascular repair of carotid artery aneurysm with Jostent covered stent: initial experience and one-year result. AB - Treatment for symptomatic extracranial carotid artery aneurysm is evolving and we describe two cases of successful endovascular treatment of distal internal carotid aneurysm using Jostent, a balloon-expandable coronary polytetrafluoroethylene-covered stent. Both patients remained symptom-free at 1 year after the procedure and patency of the stents was maintained. We conclude that endovascular treatment of carotid artery aneurysm with Jostent is feasible and safe and may be more preferable than conventional strategies. PMID- 15343563 TI - Optimal therapeutic approaches to femoropopliteal artery intervention. AB - Superficial femoral artery disease presents a complex challenge for therapy. The extent of vascular involvement may vary from focal disease with symptoms of intermittent claudication to long total occlusions manifest as critical limb ischemia. Optimal therapy requires understanding the available options including exercise programs, pharmacologic medical therapy, surgery and interventional endovascular therapy. Rapidly advancing endovascular technology for enabling safe intervention in complex, long occlusive segments of the superficial femoral artery continues to emerge. New devices like the SafeCross wire, Excimer laser, Silverhawk Atherectomy catheter, Cryoplasty catheter and new generations of bare metal and drug-eluting nitinol stents are shifting the paradigm for therapy from surgical to more endovascular treatment even for the most complex disease presentation. PMID- 15343564 TI - Angiographic adverse events during percutaneous coronary intervention fail to predict creatine kinase-MB elevation. AB - We attempted to determine if aggressive detection of angiographic adverse events during coronary intervention could predict subsequent creatine kinase (CK)-MB elevations. During coronary intervention, both fluoroscopy and cine angiography were used to detect angiographic adverse events. At least one angiographic adverse event occurred in 133/251 (53%) of procedures. CK-MB elevation occurred in 24% of procedures. Slow flow during the procedure (P=0.002) and chest discomfort at the end of the procedure (P=0.007) were the strongest predictors of CK-MB elevation. Among procedures with no angiographic adverse events, CK-MB elevation occurred in 15/121 (12%), accounting for 25% of CK-MB elevations. We conclude that CK-MB elevation occurs after angiographically uncomplicated coronary interventions even when angiographic adverse events are aggressively detected. Routine monitoring of cardiac enzymes is necessary to detect all patients who will experience myocardial injury after coronary intervention. PMID- 15343565 TI - Invisible showers. PMID- 15343566 TI - Balloon angioplasty plus cilostazol administration versus primary stenting of small coronary artery disease: final results of COMPASS. AB - Efficacy of primary stenting in small coronary artery disease is still controversial. Cilostazol has been reported to control restenosis after balloon angioplasty (BA). The aim was to compare primary stenting with BA plus cilostazol administration in small coronary artery disease. Of 106 lesions located in small coronary artery (reference < 3.0 mm), 50 lesions were randomly assigned to the stenting and 56 lesions to the BA-cilostazol group. Multilink stent was implanted in the stenting group. In the BA-cilostazol group, cilostazol (200 mg/day) without aspirin was administered for 6 months after BA. Ticlopidine was given for 1 month when bailout stent was implanted. Serial quantitative angiography was performed at the procedure and 6 months. The primary endpoint was 6-month angiographic restenosis. Clinical event rates at 1 year were also assessed. Baseline characteristics were similar. All procedures were successful. Bailout stenting was performed in three lesions in the BA-cilostazol group. No side effects of cilostazol were observed. Postprocedural lumen diameter was significantly larger (2.69 vs. 2.03 mm; P < 0.0001) in the stenting group. However, the follow-up lumen diameter was not different (1.76 vs. 1.85 mm, stenting vs. BA-cilostazol). Although the difference was not statistically significant, restenosis rate was lower in the BA-cilostazol group (13.2% vs. 24.5%; P = 0.11). Subacute thrombosis occurred in one patient and target revascularization rate was higher in the stenting group (22.0% vs. 10.7%; P = 0.10). BA plus cilostazol administration seems to be a favorable strategy for small coronary artery disease. PMID- 15343567 TI - Contribution of early lumen loss after balloon angioplasty for in-stent restenosis to lumen loss at follow-up. AB - The treatment of in-stent restenosis using balloon angioplasty alone often produces excellent early results, but is associated with high rate of recurrence. Previous studies have demonstrated significant tissue reintrusion shortly after the treatment of in-stent restenosis with balloon angioplasty. The study was designed to elucidate the contribution of early lumen loss 6 hr after balloon angioplasty to lumen loss at follow-up. We prospectively performed quantitative coronary angiography and intravascular ultrasound in 12 patients with in-stent restenosis before intervention, after the final procedure, 6 hr later (5.6 +/- 1.4 hr), and at follow-up (7.7 +/- 2.3 months). Compared with immediately after balloon angioplasty, by 6 hr postintervention, the minimum lumen diameter (MLD) and lumen cross-sectional area had decreased significantly (2.48 +/- 0.44 to 2.01 +/- 0.57 mm, P = 0.01, and 7.0 +/- 1.2 to 5.5 +/- 1.4 mm2, P = 0.004, respectively). Furthermore, the MLD decreased further between 6 hr postintervention and long-term follow-up (2.01 +/- 0.57 to 1.55 +/- 0.64 mm; P = 0.001). Patients who showed recurrence of restenosis at follow-up had greater early lumen loss than patients without recurrence of restenosis (0.71 +/- 0.31 vs. 0.23 +/- 0.13 mm; P = 0.006). Diffuse lesions had greater early lumen loss compared to focal lesions (0.75 +/- 0.35 vs. 0.28 +/- 0.13 mm; P = 0.008). Early lumen loss is common after the treatment of in-stent restenosis by balloon angioplasty. Within the first 6 hr postintervention, 32% +/- 29% of acute lumen gain is lost, and early lumen loss contributed to 42% +/- 18% of total lumen loss at follow-up. PMID- 15343568 TI - Elective sirolimus-eluting stent implantation for multivessel disease involving significant LAD stenosis: one-year clinical outcomes of 99 consecutive patients- the Rotterdam experience. AB - The aim of this study was to evaluate the effectiveness of sirolimus-eluting stent (SES) implantation for patients with multivessel disease, which included left anterior descending artery (LAD) treatment. Since April 2002, SES has been utilized as the device of choice for all interventions in our institution as part of the Rapamycin-Eluting Stent Evaluated at Rotterdam Hospital (RESEARCH) registry. In the first 6 months of enrollment, 99 consecutive patients (17.6% of the total population) were treated for multivessel disease involving the LAD. The impact of SES implantation on major adverse cardiac events (MACE) was evaluated. All the patients received SES in the LAD. Additional stent implantation in the right coronary artery, the left circumflex, or in all three major vessels was attempted successfully in 32 (32%), 51 (52%), and 16 (16%) of the treated patients respectively. During a mean follow-up of 360 +/- 59 days (range, 297-472 days), we had one death, one non-Q-wave myocardial infarction, and eight patients required subsequent intervention. The event-free survival of MACE at 1 year was 85.6%. SES implantation for multivessel disease in a consecutive series of patients is associated with low incidence of adverse events. The reported results are related predominantly to the reduction in repeat revascularization. PMID- 15343569 TI - Single-digit target vessel revascularization in multivessel coronary interventions with sirolimus-eluting stents: goodbye to coronary bypass grafting. PMID- 15343570 TI - Catheter-based intracardiac echocardiography in the interventional cardiac laboratory. AB - Recent advances in technology have engendered a renewed enthusiasm in the use of intracardiac echocardiography (ICE) to guide and assess cardiac interventions. AcuNav is a phased-array sector imaging probe equipped with color and spectral Doppler capabilities. Previous-generation imaging catheters yielded unfamiliar limited-depth radial images with no flow information. Current imaging technology such as the AcuNav has not only consolidated the role of ICE but opened newer applications in the interventional laboratory. ICE has clear advantages over transesophageal echocardiography as the imaging modality of choice in the cardiac catheterization and electrophysiological laboratories. We review the technical evolution of ICE and describe the expanded utility of the AcuNav imaging catheter during cardiac interventions. PMID- 15343571 TI - Transthoracic access for cardiac catheterization. AB - Patients who have undergone Fontan palliation for congenital heart disease have limited access to the pulmonary venous atrium. We describe a transthoracic approach to the pulmonary venous atrium for diagnostic and therapeutic cardiac catheterization. A total of 22 patients ranging in age from 14 months to 25 years and weighing 7-68 kg underwent transthoracic catheterization. With the aid of fluoroscopic and angiographic markers, a 20 gauge needle was inserted into the pulmonary venous atrium. A floppy-tipped wire was inserted, and a 4 Fr sheath was placed, upsized as needed. Access to the pulmonary venous atrium was obtained for all patients. Therapeutic procedures were completed for all patients. Transthoracic access to the pulmonary venous atrium in patients who have undergone a Fontan operation is a reliable and rapid mode of access, allowing for complex therapeutic procedures to be performed in the catheterization laboratory, obviating the need for additional open heart surgery. PMID- 15343572 TI - Everything old is new again: transthoracic cardiac catheterization. PMID- 15343573 TI - Stent implantation for coarctation of aorta caused by a Rashkind patent ductus arteriosus umbrella. AB - This case demonstrates a novel use of stent implantation for relief of coarctation of the aorta caused by protrusion of a Rashkind patent ductus arteriosus umbrella. Follow-up 3 years after stent implantation shows complete relief of obstruction. PMID- 15343574 TI - Transcatheter closure of fistula between the right pulmonary artery and left atrium using the Amplatzer duct occluder. AB - A congenital fistula between the right pulmonary artery and the left atrium is a rare condition that results in cyanosis. We report a successful catheter-based closure of such a fistula in a 12-year-old using an Amplatzer duct occluder. The patient also had an atrial septal defect that was closed with an Amplatzer septal occluder. PMID- 15343575 TI - What will be left for the surgeons? PMID- 15343576 TI - Percutaneous stent-mounted valve for treatment of aortic or pulmonary valve disease. AB - The objective of this study was to develop a prosthetic cardiac valve designed for percutaneous transcatheter implantation. Percutaneous catheter-based therapies play a limited role in the management of cardiac valve disease. Surgical implantation of prosthetic valves usually requires thoracotomy and cardiopulmonary bypass. The stent-valve is constructed of a rolled sheet of heat treated nitinol. Although malleable when cooled, once released from a restraining sheath at body temperature the stent unrolls, becomes rigid, and assumes its predetermined cylindrical conformation. A ratcheting lock-out mechanism prevents recoil and external protrusions facilitate anchoring. Valve leaflets are constructed of bovine pericardium. The feasibility of catheter implantation, prosthetic valve function, and survival were investigated in an animal model. In vitro and pulse duplicator testing documented valve durability. Endovascular delivery of the prototype stent-valve to the aortic or pulmonary position was feasible. Accurate positioning was required to ensure exclusion of the native valve leaflets and, in the case of the aortic valve, to avoid compromise of the coronary ostia or mitral apparatus. Oversizing of the stent in relation to the valve annulus was desirable to facilitate anchoring and prevent paravalvular insufficiency. Stent-valve implantation proved feasible and compatible with survival in an animal model. Transcatheter implantation of prosthetic valves is possible. Further evolution of this technology will involve lower-profile devices with design features that facilitate vascular delivery, visualization, positioning, deployment, and valvular function. PMID- 15343577 TI - Evaluation and treatment of abnormalities of the interatrial septum. PMID- 15343578 TI - Bifurcation disease: optimal angiographic result yet suboptimal long-term outcome. PMID- 15343579 TI - Nonhealing wound resulting from a foreign body to a radial arterial sheath and sterile inflammation associated with transradial catheterization and hydrophilic sheaths. PMID- 15343582 TI - 'Lipoproteins, glycoxidation and diabetic angiopathy'. AB - The chronic vascular complications of diabetes (nephropathy, retinopathy and accelerated atherosclerosis) are a major cause of morbidity and premature mortality. In spite of the more widespread availability of intensive diabetes management, approximately one in three people with diabetes develop aggressive complications and over 70% die of atherosclerosis-related diseases. Genetic and acquired factors are likely to be contributory. Potential mediators of vascular damage may include the interrelated processes of lipoprotein abnormalities, glycation, oxidation and endothelial dysfunction. Lipoprotein abnormalities encompass alterations in lipid concentrations, lipoprotein composition and subclass distribution and lipoprotein-related enzymes. Nonenzymatic glycation and oxidative damage to lipoproteins, other proteins and to vascular structures may also be deleterious. As atherosclerosis is a chronic condition commencing in youth, and because clinical events may be silent in diabetes, surrogate measures of vascular disease are important for early identification of diabetic patients with or at high risk of vascular damage, and for monitoring efficacy of interventions. The increasing array of biochemical assays for markers and mediators of vascular damage, noninvasive measures of vascular health, and therapeutic options should enable a reduction in the excessive personal and economic burden of vascular disease in type 1 and type 2 diabetes. PMID- 15343583 TI - Fructose-mediated non-enzymatic glycation: sweet coupling or bad modification. AB - The Maillard reaction is a process in which reducing sugars react spontaneously with amino groups in proteins to advanced glycation end products (AGEs). Although an elevated level of glucose had been thought to play a primary role in the Maillard reaction, on a molecular basis, glucose is among the least reactive sugars within biological systems. The formation of AGEs is now also known to result from the action of various metabolites other than glucose, which are primarily located intracellularly and participate in the non-enzymatic glycation reaction at a much faster rate, such as fructose, trioses and dicarbonyl compounds. In this review, we considered the glycation reaction with particular attention to the potential role of fructose and fructose metabolites. The two sources for fructose are an exogenous supply from the diet and the endogenous formation from glucose through the aldose reductase pathway. Despite its approximately eightfold higher reactivity, the contribution of extracellular glycation by fructose is considerably less than that by glucose, because of the low plasma concentration of fructose (5 mmol/L glucose vs 35 micro mol/L fructose). Intracellularly, fructose is elevated in a number of tissues of diabetic patients in which the polyol pathway is active. In the cells of these tissues, the concentrations of fructose and glucose are of the same magnitude. Although direct evidence is not yet available, it is likely that the high reactivity of fructose and its metabolites may substantially contribute to the formation of intracellular AGEs and may contribute to alterations of cellular proteins, dysfunction of cells and, subsequently, to vascular complications. PMID- 15343584 TI - Exercise as a therapeutic intervention for the prevention and treatment of insulin resistance. AB - During the past half-century, there has been a dramatic increase in the incidence of obesity, insulin resistance and type 2 diabetes. A hallmark feature of these conditions is impaired glucose tolerance (IGT). Cross-sectional and retrospective epidemiological studies have provided direct evidence that a lack of physical activity is strongly associated with IGT. Indeed, physical inactivity is an independent risk factor for insulin resistance and type 2 diabetes. The primary defect(s) in the development of whole-body insulin resistance remain unclear. However, during the past decade significant progress has been made towards an understanding of the molecular basis underlying the beneficial effects of exercise training in stimulating the entry of glucose into insulin-sensitive tissues. Accordingly, it is now well accepted that regular physical exercise offers an effective therapeutic intervention to improve insulin action in skeletal muscle in insulin-resistant individuals. This review provides evidence that physical inactivity is significantly associated with IGT and directly contributes to the cascade of events that lead to the expression of the 'exercise deficient phenotype' associated with insulin resistance and type 2 diabetes. In contrast, exercise training will be shown to significantly reduce the risk of developing insulin resistance by improving glucose tolerance and insulin action in individuals predisposed to develop type 2 diabetes. Several putative mechanisms for enhanced glucose uptake after exercise training will be discussed. A determination of the underlying biological mechanisms that result from exercise training is essential in order to define the precise variations in physical activity that result in the most desired effects on targeted risk factors, and to aid in the development of such interventions. PMID- 15343585 TI - The natural history of insulin content in the pancreas of female and male non obese diabetic mouse: implications for trials of diabetes prevention in humans. AB - BACKGROUND: The non-obese diabetic (NOD) mouse is a well-established animal model used to study the pathogenesis of type 1 diabetes. The NOD mouse spontaneously develops an autoimmune form of the disease between 12 and 18 weeks of age, characterized by an infiltration of the endocrine pancreas by autoreactive mononuclear cells. In our colony, all animals showed signs of insulitis, but only approximately 60% of females and 15% of males developed diabetes. The aim of this study was to determine the natural history of insulin content in the pancreas of female and male NOD/Ba mice during their life span. METHODS: Pancreata were collected at two-week intervals, from 4 weeks of age to 30 weeks of age. Four animals at each age as well as 18 diabetic female NOD mice were studied. Pancreata were homogenized, supernatants collected and insulin measured by radioimmunoassay. RESULTS: Female and male non-diabetic NOD mice showed significantly higher levels of insulin in the pancreata in comparison to the diabetic female animals. Pancreata from female (n = 56) animals showed more insulin content than male pancreata (n = 56), suggesting beta-cell hyperactivity as a result of the ongoing beta-cell destruction. However this difference was only significant at an early age (4-12 weeks of age) (p < 0.04). Insulin content in diabetic female NOD pancreas declines with time, and was very low at the age of 25 to 34 weeks. This decline was not observed in male pancreata despite the presence of lymphocytic infiltration. CONCLUSION: We conclude that a reduction in pancreatic insulin content occurs slowly in the natural history of the disease and that such reduction only becomes apparent after diagnosis of hyperglycaemia. Occurrence of extensive lymphocytic infiltration in non-diabetic male mice is not accompanied by a reduction of insulin content in the pancreas. These findings have implications for designing studies in humans which aims to protect residual beta-cell function. PMID- 15343586 TI - Increasing of oxidative stress from mitochondria in type 2 diabetic patients. AB - BACKGROUND: Recent evidence increasingly indicates that oxidative stress may play an important role in the pathogenesis of diabetic vascular complications. Mitochondria has received much attention as an important organ in the generation of oxidative stress. However, the importance of oxidative stress among diabetic patients without vascular complications is unclear. METHODS: We compared oxidative stress produced from mitochondria of the mononuclear cells in peripheral blood obtained from 26 diabetic subjects without clinical vascular complications and 52 healthy age-matched subjects using a flow cytometer. Oxidative stress from the mononuclear cells was evaluated by measuring fluorescence of oxidized production from dihydrorhodamine-123, which is a pro fluorescent compound that selectively accumulates in the mitochondria of living cells. Stimulation of the cells was carried out with phorbol 12-myristate 13 acetate (PMA), a protein kinase C (PKC) activator. We then calculated the relative fluorescence variation (RFV) that indicated an increasing rate of oxidative stress levels by stimulation with PMA against the levels obtained at baseline. Additionally, we measured the urinary stress markers, 8 hydroxydeoxyguanosine (8OHdG) and 8-epi-prostaglandin F2alpha (isoprostane). RESULTS: Compared to healthy subjects, diabetic subjects did not exhibit significantly elevated oxidative stress levels at baseline, but did have significantly elevated basal urinary 8OHdG, urinary isoprostane and oxidative stress levels after PMA stimulation as well as RFV. CONCLUSIONS: Among diabetic subjects without clinical vascular complications, there was a possibility that mitochondrial oxidative stress balance between generation and scavenging against the additive PKC stimulation was thought to have already been lost. PMID- 15343587 TI - Effects of insulin-induced hypoglycaemia on energy intake and food choice at a subsequent test meal. AB - BACKGROUND AND AIMS: Hypoglycaemia is assumed to increase food intake, but there is little data on the magnitude or qualitative nature of this effect. We have therefore investigated the effects of insulin-induced hypoglycaemia on food intake at a test meal. METHODS: Sixteen healthy men (age 29.8 +/- 11 years; mean +/- SD) were studied; either insulin (0.05 units/kg) or saline was given intravenously in a double-blind crossover design. Blood glucose was monitored at regular intervals. Participants were given an ad libitum breakfast 20 min after injections and food intake and appetite scores were recorded. RESULTS: Blood glucose was unchanged following saline (4.3 +/- 0.4 to 4.4 +/- 0.3 mmol/L). There was a transient decline in blood glucose after insulin with a nadir at 20 min (4.31 +/- 0.34 to 2.41 +/- 0.45 mmol/L, p < 0.0001), which returned to baseline at 40 min. Total energy intake was 17% higher (1701.1 +/- 895.3 kcal vs 1427.7 +/ 815 kcal, p = 0.026) following insulin administration compared to that following saline. Macronutrient analysis revealed a significant increase in high-fat foods (muffins) (69.2 +/- 54.1 vs 29 +/- 42.3 g, p = 0.009) after insulin. Appetite scores were similar after saline and insulin despite these changes in food intake. CONCLUSIONS: Transient insulin-induced hypoglycaemia increases energy intake. Participants consumed more fat after insulin compared to that after saline. High-fat foods can lead to passive overconsumption and have a low glycaemic index, which may prolong hypoglycaemia. Both factors could ultimately promote weight gain in individuals with recurrent hypoglycaemia. PMID- 15343588 TI - Effectiveness of progressive dose-escalation of exenatide (exendin-4) in reducing dose-limiting side effects in subjects with type 2 diabetes. AB - BACKGROUND: Exenatide (exendin-4) exhibits dose-dependent glucoregulatory activity, but causes dose-limiting nausea and vomiting. This study was designed to formally assess the possibility of inducing tolerance to the side effects of nausea and vomiting at therapeutic doses of exenatide, using a dose-escalation methodology. METHODS: In this two-arm, triple-blind, multicenter study, 123 subjects with type 2 diabetes were enrolled and randomized; 99 (80.5%) of them completed the study. Subjects in the exenatide-primed arm received subcutaneous exenatide, starting at 0.02 micro g/kg three times a day (TID) and increasing in 0.02 micro g/kg per dose increments every 3 days for 35 days. Subjects in the exenatide-naive arm received placebo TID for 35 days. At the end of this 35-day regimen, subjects in both arms received the same highest dose of exenatide (0.24 micro g/kg TID) for 3 days. Thus, the exenatide-naive arm received exenatide for the first time on Day 35. RESULTS: The exenatide-primed arm had a lower proportion of subjects experiencing nausea and vomiting in response to exposure to the highest dose of exenatide (27 vs 56% in the exenatide-naive arm; p = 0.0018). Kaplan-Meier estimates of cumulative incidence were 0.28 in the exenatide-primed arm, compared with 0.68 in the exenatide-naive arm (p or = 4 mm, 142 fetuses with trisomy 21, 58 with trisomy 18, 19 with trisomy 13, 37 with Turner's syndrome and 24 with isolated heart defects. Groups were compared using Student's t-test and confidence intervals for differences between groups were calculated. RESULTS: Otherwise normal fetuses with increased NT showed no difference in any of the cardiac Doppler parameters from normal controls. Mean E/A ratio was slightly but significantly increased in trisomy 21 fetuses compared with normal controls (0.604 vs. 0.578 on the right, P = 0.011; 0.581 vs. 0.542 on the left, P = 0.0001). E/A ratio was not significantly different between any of the other groups and the normals but there was a small increase in absolute E-wave velocity in trisomy 18 fetuses. MPI was significantly decreased in trisomy 21 fetuses, (0.330 vs. 0.378, P = 0.002 on the left) and also in Turner's syndrome fetuses (0.301 vs. 0.352 on the right, P = 0.04; 0.320 vs. 0.378 on the left, P = 0.034) implying better performance, but not in the other groups. CONCLUSIONS: The magnitude and/or direction of the differences shown do not support a major role for cardiac functional abnormality in the development of NT. Important cardiac dysfunction could not be demonstrated in association with increased NT in normal or abnormal fetuses. PMID- 15343593 TI - Fetal gender and sonographic assessment of crown-rump length: implications for multifetal pregnancy reduction. AB - OBJECTIVES: Fetal size is a common criterion for the selection of an individual fetus to be reduced during multifetal pregnancy reduction. We investigated whether a difference in crown-rump length (CRL) exists between male and female fetuses at 9-13 weeks' gestation. METHODS: A total of 883 singleton pregnancies was evaluated at the time of chorionic villus sampling. The mean gestational age at the time of intervention was 10.9 weeks. Pregnancies were dated by certain last menstrual period (LMP). Pregnancies with uncertain dating by LMP or with a sonographic difference of gestational age > 1 week compared with LMP were excluded, as were all cases with chromosomal abnormalities. CRL differences between male and female fetuses were compared using Student's t-test. RESULTS: A total of 417 female and 466 male fetuses fulfilled our study criteria. Their gestational ages ranged between 9.3 and 13.9 weeks. We found no significant difference in size between the male and female fetuses. CONCLUSION: Using the CRL to guide multifetal pregnancy reduction should not result in a clinically significant selection of either gender. PMID- 15343594 TI - Two- and three-dimensional sonographic assessment of the fetal face. 2. Analysis of cleft lip, alveolus and palate. AB - OBJECTIVES: To describe the sonographic appearance of cleft lip with or without cleft palate (CL +/- P) using two-dimensional and three-dimensional (3D) ultrasound imaging. Also, to evaluate the accuracy of ultrasound to delineate with precision the bony extent of facial clefts, i.e. to differentiate clefts limited to the lips, or extending to the alveolus/premaxilla or the secondary palate. METHODS: This was a retrospective study based on the examination of fetuses diagnosed with an isolated CL +/- P. Cases included were either discovered at systematic screening or referred for further investigation. Clefts were characterized by their precise anatomical location and extent. The defect could include a cleft lip (CL), a cleft alveolus (CA), or a cleft of the secondary palate (CSP). RESULTS: We analyzed 96 cases of CL +/- P. The mean gestational age at examination was 28.2 +/- 4.1 weeks. The sonographic appearance of CL, CA, and CSP was depicted. Strict concordance of the sonographic report with the anatomical defect was present in 84 cases (87.5%). In eight cases, the severity of the cleft was underestimated: three cases of CA, four of CA + CSP and one of CSP were missed. In four cases, the cleft was overestimated as CA was incorrectly suspected. CONCLUSIONS: Systematic screening with sonography to detect prenatally CL +/- P requires the imaging of at least the mid-sagittal and the anterior coronal 'nose-mouth' views. Once the presence of a facial cleft is suspected, the three reference orthogonal planes are imaged in order to characterize the anatomical defect, and for each plane, the serial scans are thoroughly examined. This protocol allows precise delineation of the defect. Inclusion of 3D and 4D ultrasound imaging in the examination protocol allows easier and more rapid screening and more precise evaluation of the different cleft constituents. PMID- 15343595 TI - The role of three-dimensional ultrasound in visualizing the fetal cranial sutures and fontanels during the second half of pregnancy. AB - OBJECTIVES: The aim of this study was to evaluate the significance of three dimensional (3D) ultrasound in visualizing fetal cranial sutures and fontanels and to determine factors that could influence visualization and image quality. METHODS: Serial 3D ultrasound examinations were evaluated for visibility of fetal cranial sutures and fontanels, image quality and possible influencing parameters in the second half of pregnancy. Thirty fetuses were scanned at four different gestational ages providing a data set of 120 cases. RESULTS: Most (82-100%) cranial sutures and fontanels could be visualized with 3D ultrasound. However, the sagittal suture and posterior fontanel were visualized in only 47% and 42%, respectively. Gestational age significantly influenced the visibility of the sutures and fontanels, image quality decreasing with advancing gestational age. CONCLUSIONS: 3D ultrasound can be a reliable technique for visualizing most fetal cranial sutures and fontanels. By performing a sagittal and a transverse scan, most of the sutures and fontanels can be made visible during the second half of pregnancy. Visualization depends on gestational age. PMID- 15343596 TI - Congenital curly toe of the fetus. AB - OBJECTIVES: To assess the prevalence, prenatal detection rate, and associated anomalies of congenital curly toe in an unselected obstetric population. METHODS: Between September 2001 and July 2002, 1167 singleton fetuses in the second and third trimesters underwent routine prenatal ultrasound at our hospital. Congenital curly toe was diagnosed when the fourth or fifth toe was not fully delineated on the axial image with medial and plantar flexion on the coronal image. All neonates underwent a physical examination within 3 days after delivery. Radiography was performed on those infants with curly toe. We assessed the prevalence, prenatal detection rate, and associated anomalies of the condition. RESULTS: There were 38 congenital curly toes among the 1167 neonates, yielding a total prevalence of 32.6 per 1000. In 26 of the 38 the fourth toe was curly and in 12 the fifth toe was curly. Of those in which the fourth toe was curly, 26.9% involved the right toe, 65.4% the left, and 7.7% involved both toes. For a curly fifth toe, these values were 25.0%, 16.7% and 58.3%, respectively. There was no associated structural anomaly, chromosomal abnormality, or syndrome in any case. Sixteen of the 38 curly toes were detected on prenatal ultrasound, including 13 of 26 with the fourth toe being curly and three of 12 with the fifth being curly. CONCLUSIONS: Congenital curly toe is not an uncommon condition, and can be detected on the prenatal ultrasound. Although it usually appears as an isolated finding without clinical significance, thorough ultrasound examination of the fetus should be recommended. PMID- 15343597 TI - Intrapartum sonography to determine fetal occipital position: interobserver agreement. AB - OBJECTIVE: To investigate the interobserver agreement on the intrapartum ultrasonographic definition of the fetal occipital position. METHODS: In 60 singleton pregnancies in labor at term the fetal occipital position was determined by transabdominal ultrasound by two appropriately trained sonographers who were not aware of each other's findings. The Bland-Altman plot was performed and the 95% limits of agreement were calculated. Logistic regression analysis was used to investigate the association between complete agreement in the fetal occipital position between the two observers and maternal and labor characteristics. RESULTS: The two observers had complete agreement on the fetal occipital position in 22/60 (36.7%) cases and disagreement by 15 degrees and 30 degrees in 31 (51.7%) and seven (11.6%) cases, respectively. The mean of the differences between the two observers was 0.25 degrees and the 95% limits of agreement were -28.9 degrees (-32.2 degrees to -25.6 degrees) to 29.4 degrees (26.1 degrees to 32.7 degrees). There were no significant associations between complete agreement and maternal and labor characteristics. CONCLUSION: The interobserver agreement on sonographically determined fetal occipital position during labor is within 15 degrees in nearly 90% of cases and within 30 degrees in all cases. PMID- 15343598 TI - Investigation of occiput posterior delivery by intrapartum sonography. AB - OBJECTIVE: To investigate if occiput posterior delivery is the consequence of persistence of an initial occiput posterior position or malrotation from an initial occiput anterior or transverse position. METHODS: This was a cross sectional study involving transabdominal sonography to determine fetal occipital position in 918 singleton pregnancies with cephalic presentation in active labor at 37-42 weeks of gestation. The relationship between occipital position in labor and at delivery was examined. RESULTS: The occiput was posterior in 33.0% (149/452), 33.9% (101/298) and 19.0% (32/168) of fetuses at the respective cervical dilatations of 3-5, 6-9 and 10 cm and this persisted at delivery in 21.5% (32/149), 31.7% (32/101) and 43.8% (14/32) of cases. In 70% (32/46), 91% (32/35) and 100% (14/14) of occiput posterior deliveries there was persistence from this position at 3-5, 6-9 and 10 cm of cervical dilation. CONCLUSIONS: The majority of occiput posterior positions during labor rotate to the anterior position even at 10 cm of cervical dilatation. However, the vast majority of occiput posterior positions at delivery are a consequence of persistence of this position during labor rather than malrotation from an initial occiput anterior or transverse position. PMID- 15343599 TI - Clinical significance of uterine artery blood flow velocity waveforms during provoked uterine contractions in high-risk pregnancy. AB - OBJECTIVE: To determine whether, in a group of high-risk pregnancies undergoing an oxytocin challenge test (OCT), uterine artery Doppler velocimetry will identify fetuses at risk of distress during the provoked contractions. METHODS: Bilateral uterine artery Doppler velocimetry was performed simultaneously with electronic fetal heart rate (FHR) recordings in 67 high-risk pregnancies subjected to an OCT. Flow velocity waveforms (FVWs) were classified according to pulsatility index (PI), presence of diastolic notching and a novel classification of FVW shapes. The OCT was classified as negative (normal) or positive (late FHR decelerations). Only OCT-negative cases were allowed a trial of vaginal delivery. Non-parametric statistical methods were used to test for differences between the OCT groups. RESULTS: There was no difference in prevalence of high PI or diastolic notching in OCT-positive (n = 10) and OCT-negative (n = 57) cases at basal (resting) measurements or between uterine contractions in either uterine artery (P > or = 0.3). During contractions the PI could not be used for assessment due to the biphasic shape of the FVWs, but there was no difference in distribution of FVW classes between the groups in either the placental side (P > or = 0.3) or contraplacental side (P > or = 0.6) uterine artery. No significant associations between PI or FVW class distribution and birth asphyxia or operative delivery for fetal distress in labor were found (P > or = 0.1). CONCLUSIONS: During uterine contractions there is no difference in uterine artery FVW pattern between OCT-positive and OCT-negative cases. Recording of uterine artery FVWs during the OCT seems to be of limited clinical relevance. PMID- 15343600 TI - Second-trimester uterine artery Doppler and spontaneous preterm delivery. AB - OBJECTIVE: To assess the relationship between second-trimester uterine artery (UtA) Doppler measurements and spontaneous preterm delivery. METHODS: This was a retrospective analysis of UtA Doppler findings at 18-23 weeks' gestation in 234 singleton pregnancies with spontaneous preterm labor and 5472 pregnancies delivered at term. Cases with fetal chromosomal or structural abnormalities, pre eclampsia, small-for-gestational-age (SGA) fetuses, concurrent maternal disease, cervical cerclage or history of cone biopsy were excluded. RESULTS: UtA mean resistance index (RI) and number of protodiastolic notches were not significantly different in the spontaneous preterm labor group compared to the normal group. The 62 preterm deliveries not preceded by spontaneous preterm premature rupture of membranes (PPROM) showed a significantly higher RI and prevalence of bilateral notches than cases of preterm deliveries preceded by PPROM (n = 172). However, this difference was not confirmed in a logistic regression model. CONCLUSION: Despite pathological evidence suggesting that defective placentation is associated with spontaneous preterm delivery, second-trimester UtA resistance, as assessed by Doppler ultrasound investigation, is not different in pregnancies subsequently complicated by preterm labor compared to pregnancies delivered at term. PMID- 15343601 TI - Ultrasound training: the virtual patient. AB - OBJECTIVE: To evaluate an ultrasound training system designed to standardize teaching and learning of gynecological sonography using a virtual model. METHODS: The 'virtual patient' was based on a three-dimensional freehand ultrasound system that allows two-dimensional sonographic offline investigations of previously recorded cases, imitating a real gynecological scan. In the first test phase designed to check the congruence of real and virtual scans, 25 doctors experienced in ultrasound examined three virtual cases. During the second test phase we assessed whether training with the virtual patient helped to establish a satisfactory practical knowledge of gynecological ultrasound. This phase was carried out with 24 medical students without ultrasound experience. RESULTS: All 25 doctors successfully investigated the three cases and generated an accurate diagnosis for the first and second cases. In the third case 14 doctors made the correct diagnosis (uterus bicornis). The measurements of endometrial thickness and the diameter of a fibroid yielded acceptable results compared with the original investigation. After a short standardized video-based instruction, all 24 medical students were able to perform a basic transvaginal scan and to inspect the uterus, ovaries and the urinary bladder. Measurements of endometrial thickness by all students deviated minimally from the actual measurement. CONCLUSIONS: Training with the virtual patient appears to be comparable to performing a live gynecological ultrasound investigation and allows standardized ultrasound teaching and learning. PMID- 15343602 TI - Color Doppler velocimetry and three-dimensional color power angiography of cervical carcinoma. AB - OBJECTIVES: To investigate the blood flow within invasive cervical carcinoma by transvaginal two-dimensional (2D) color spectral Doppler and three-dimensional (3D) color power angiography and to correlate these parameters with the clinicopathological characteristics. METHODS: Seventy-four patients with invasive cervical carcinoma were enrolled for the analysis. Squamous cell carcinoma serum antigen levels (SCC) were obtained for all the patients. Sections of all malignant tissues were analyzed for tumor expression of cyclooxygenase-2 (COX-2). All patients underwent color and spectral Doppler examination and 44 patients had 3D color power angiography. Color spectral Doppler parameters (color score, lowest resistance index (RI), highest peak systolic velocity (PSV)) and 3D color power angiography indices (relative color, average color, flow measure) of FIGO I/II cervical cancers were compared with those obtained in a control group of 24 patients with a normal uterine cervix. Pulsed Doppler parameters and the 3D vascular indices were compared with clinicopathological parameters, SCC serum antigen levels and tumor COX-2 expression. RESULTS: At color Doppler analysis 72 patients (97%) showed intralesional detectable vessels. Color spectral Doppler and 3D-derived parameters were significantly different in FIGO I/II cervical cancers compared with those in women with a normal cervix. A significantly higher color score (P = 0.0008), lower RI (P = 0.032) and higher PSV (P = 0.004) were associated with a tumor diameter > or =4 cm compared with smaller tumors. The highest PSV was significantly higher in patients with FIGO stage III/IV compared with FIGO stage I/II (P = 0.0069). There was a direct correlation between PSV and SCC (r = +0.44, P = 0.003). The median relative color was significantly higher in patients with a higher color score (P = 0.0006). No statistically significant correlations were found between 3D color power angiography parameters and the clinicopathological characteristics or between the 3D vascular parameters and biological factors. CONCLUSIONS: Alterations of 3D ultrasound-derived vascular indices were found in patients with cervical cancer compared with those with a normal cervix. Moreover, some vascular indices proved to be associated with tumor size. The assessment of a possible clinical role of 2D and 3D ultrasound-derived vascular indices in cervical cancer deserves further investigation. PMID- 15343603 TI - Endometrial development in association with ovarian follicular waves during the menstrual cycle. AB - OBJECTIVES: Waves of ovarian follicular development during the menstrual cycle have recently been documented in our laboratory. The objective of this study was to test the hypothesis that ultrasonographically detectable changes in the endometrium during the menstrual cycle would differ between women with two vs. three waves of ovarian follicular development and among women with different major and minor wave patterns of follicle growth. METHODS: Fifty women of reproductive age (mean age +/- SD, 28.0 +/- 6.9 years) underwent daily transvaginal ultrasonography for one interovulatory interval (IOI). Ultrasonographic images of the endometrium were obtained each day, and measurements of endometrial area and perimeter (based on the shape of an ellipse, in the transverse plane) and thickness and pattern (in the sagittal plane) were recorded. Endometrial area, perimeter, thickness and pattern were compared between women with two and three waves of follicle development and among women with different minor and major wave patterns of follicular growth during the IOI. RESULTS: Endometrial area, perimeter, thickness and pattern increased earlier during the follicular phase in women with two vs. three waves of follicular development. In women with two follicle waves, endometrial area and perimeter increased earlier in those with major major vs. minor major follicle wave patterns. CONCLUSIONS: Ultrasonographically detectable changes in the endometrium occurred in association with follicle wave dynamics in women. Earlier development of the endometrium during the follicular phase in women with two vs. three follicle waves was attributed to an earlier increase in dominant follicle estradiol production. PMID- 15343604 TI - Massive multicystic dilatation of the uterine wall with myometrial venous thrombosis during pregnancy. AB - We present a pregnancy complicated by multicystic dilatation of the uterine wall during the second trimester, leading to massive uterine distension, anemia and preterm Cesarean section. The cystic changes detected by ultrasound and magnetic resonance imaging involved the whole uterine wall surrounding the entire amniotic cavity. Histopathological examination revealed the benign nature of the cystic changes, which represented dilated and thrombosed venous lacunae. Disturbed venous drainage, combined with local thrombosis, was likely to have led to the collection of a large volume of blood in the uterine wall and the subsequent multicystic change of the myometrium. PMID- 15343605 TI - Complete duplication of urinary bladder and urethra: prenatal sonographic features. AB - Prenatal sonographic features of the rare anomaly of complete duplication of the urinary bladder and urethra are described in this case report. A coronal scan of the fetal pelvis at 29 weeks of gestation revealed two pyriform cystic structures. The umbilical arteries coursed around both of them. They emptied independently of each other. Postnatally the newborn had two vulvae, two anal openings, two bladders and two uteri. PMID- 15343606 TI - Treatment of fetal heart block with maternal steroid therapy: case report and review of the literature. AB - The presence of maternal autoantibodies to SS-A/Ro and/or SS-B/La is associated with the development of fetal heart block. There are data suggesting that maternal treatment with steroids might reverse heart block. We report on a pregnancy in a mother with secondary Sjogren syndrome and systemic lupus erythematosus with presence of autoantibodies to SS-A/Ro and SS-B/La, which was complicated by the development of incomplete fetal heart block. Oral dexamethasone treatment could not prevent progression to complete heart block and was associated with a number of complications.A review of the literature revealed 19 studies (including ours) in which 93 cases of fetal heart block were treated with maternal steroid therapy. Complete heart block proved irreversible in all cases; and of 13 fetuses with incomplete heart block which received maternal steroid therapy, three had a reduction in their degree of block and one reverted to sinus rhythm. Maternal steroid therapy, initiated early in pregnancy and potentially preventing the onset of heart block, did not decrease the incidence of heart block in nine studies with 43 cases. Furthermore, the literature review revealed numerous serious side effects of maternal steroid administration during pregnancy. Data on these potential side effects are lacking in the 28 studies discussed in this review. Maternal dexamethasone therapy to prevent or treat fetal heart block remains, in our opinion, a questionable intervention and can as yet not be recommended in the clinical situation. PMID- 15343607 TI - Unsuccessful management of acardiac fetus with intrafetal alcohol injection. PMID- 15343608 TI - Lack of normalization of middle cerebral artery flow velocity prior to fetal death before the 30th week of gestation: a report of three cases. PMID- 15343609 TI - Prenatal diagnosis of lobar holoprosencephaly using color Doppler: three cases with the anterior cerebral artery crawling under the skull. PMID- 15343610 TI - Re: Intra-amniotic Doppler measurement of blood flow in placental vascular anastomoses in twin-twin transfusion syndrome. PMID- 15343612 TI - Late onset microcephaly: failure of prenatal diagnosis. PMID- 15343613 TI - Fetal heart screening--have we hit a glass ceiling? PMID- 15343614 TI - Cat-eye syndrome in a fetus with increased nuchal translucency: three-dimensional ultrasound and echocardiographic evaluation of the fetal phenotype. PMID- 15344186 TI - Two-stage adaptive strategy for superiority and non-inferiority hypotheses in active controlled clinical trials. AB - In active controlled trials without a placebo arm, there are usually two study objectives: to test a superiority hypothesis that the experimental treatment is more effective than the active control therapy, and to test a non-inferiority hypothesis that the experimental treatment is therapeutically no worse than the active control within a defined margin. For a two-stage adaptive design, it is not necessary to give a fixed sample size calculation at the planning stage of the study when treatment effect information is often insufficient. Instead, decision and estimation of the design specifications can be made more reliably after the first stage when interim results are available. We propose the use of conditional power approach to determine the sample size and critical values for testing the superiority and non-inferiority hypotheses for the second stage based on the observed result of the first stage. The proposed adaptive procedure preserves the overall type I error rate for both superiority and non-inferiority, and has the flexibility of early termination of the study (for futility or efficacy) or extending the study by appropriate sample size. PMID- 15344187 TI - Power analyses for longitudinal trials and other clustered designs. AB - Existing methods for power and sample size estimation for longitudinal and other clustered study designs have limited applications. In this paper, we review and extend existing approaches to improve these limitations. In particular, we focus on power analysis for the two most popular approaches for clustered data analysis, the generalized estimating equations and the linear mixed-effects models. By basing the derivation of the power function on the asymptotic distribution of the model estimates, the proposed approach provides estimates of power that are consistent with the methods of inference for data analysis. The proposed methodology is illustrated with numerous examples that are motivated by real study designs. PMID- 15344188 TI - Improved confidence intervals for a binomial parameter following a group sequential phase II clinical trial. AB - Group sequential procedures are widely employed in phase II clinical trials. It is often desirable to provide an interval estimate of the true response rate upon termination of a group sequential phase II clinical trial. The confidence intervals proposed by Jennison and Turnbull (Technometrics 1983; 25: 49-58) are conservative. Utilization of auxiliary statistics based on the overall disease status is proposed to reduce the discreteness of the underlying distribution. Confidence intervals generated by the proposed methods have confidence coefficients closer to the nominal level and have shorter average lengths than JT. PMID- 15344189 TI - Imputation for incomplete high-dimensional multivariate normal data using a common factor model. AB - It is common in applied research to have large numbers of variables measured on a modest number of cases. Even with low rates of missingness on individual variables, such data sets can have a large number of incomplete cases. Here we present a new method for handling missing continuously scaled items in multivariate data, based on extracting common factors to reduce the number of covariance parameters to be estimated in a multivariate normal model. The technique is compared in several simulation settings to available-case analysis and to a multivariate normal model with a ridge prior. The method is also illustrated on a study with over 100 variables evaluating an emergency room intervention for adolescents who attempted suicide. PMID- 15344190 TI - Cross calibration in longitudinal studies. AB - In a long-running longitudinal study using complex machinery to obtain measurements, it is sometimes necessary to replace the machine. This can result in lack of continuity in the measurements that can overwhelm any treatment effect or time trend. We propose a Bayesian procedure implemented using Markov chain Monte Carlo to calibrate the measurements on the old machine utilizing both person-specific and population information. The goal is to convert the previous measurements to values that can be treated as though they were made on the new machine. This methodology is applied to a bone mineral density study where the first densitometer uses gadolinium as the energy source (Lunar DP-3) and the second uses X-rays (Hologic QDR-1000W). Finally, simulation results are presented to show the superiority of the proposed method over existing methods of cross calibration. PMID- 15344191 TI - A cost-related approach for evaluating drug development programs. AB - Most pharmaceutical statisticians make a contribution to drug development at the study level by designing and analysing clinical studies. There is less involvement at the drug project level, especially when it comes to making a strategic decision for a drug development program. Motivated by the assessment of drug safety, this paper presents a new decision rule on whether or not to continue developing a new drug based on a cost function. The cost function takes into account both type I and type II errors in a decision making problem. It also incorporates both the cost and benefit from a drug development program. This cost benefit approach enables company senior management to evaluate a drug program objectively and make decisions in a quantitative and rational manner. The probabilities of making correct decisions are illustrated by using a real data example. PMID- 15344192 TI - Latent variable modelling of recovery trajectories: sexual function following radical prostatectomy. AB - As treatments improve and survival time lengthens, the course of recovery and long-term quality of life (QoL) is of greater interest. The application of latent class growth models to longitudinal QoL data provides unique insights into recovery that are not evident with marginal analyses. We examine this approach in the context of a large population-based observational study. This is the first attempt to characterize individuals' recovery over time following prostate cancer surgery, and to determine factors associated with varying recovery experiences of prostate cancer patients. Four major patterns emerged that illustrate typical patterns of recovery following radical prostatectomy. Given a man's baseline data, this method produces estimates of the probability of belonging to each recovery class. The method is presented as a useful tool for identifying hypotheses associated with recovery and potential antecedents of importance. PMID- 15344193 TI - An analysis of clustered categorical data--application in dental health. AB - Quite often in medical studies multiple discrete indicators are used to measure some characters that are defined only conceptually and are difficult to measure directly. Studies of this type exhibit categorical responses of dependent nature. Analysis of such categorical data appears to be extremely difficult (intractable) particularly in the presence of risk (causal) factors. In the present article, our purpose is to develop a latent mixture regression model for analysing such multivariate categorical data. Such a mixture model accommodates correlated and overdispersed data through the incorporation of random effects. Unfortunately, a full likelihood analysis is often hampered by the need for numerical integration. Two different procedures have been considered here. Both involve intensive computations. Numerical investigation has been carried out on the basis of a survey data covering 220 individuals from medical colleges in and around Calcutta (India). The purpose of the study is to compare tooth cleaning efficiency of brushes manufactured by different companies. PMID- 15344194 TI - Regression analysis of multiple source and multiple informant data from complex survey samples. AB - In this tutorial, we describe regression-based methods for analysing multiple source data arising from complex sample survey designs. We use the term 'multiple source' data to encompass all cases where data are simultaneously obtained from multiple informants, or raters (e.g. self-reports, family members, health care providers, administrators) or via different/parallel instruments, indicators or methods (e.g. symptom rating scales, standardized diagnostic interviews, or clinical diagnoses). We review regression models for analysing multiple source risk factors or multiple source outcomes and show that they can be considered special cases of generalized linear models, albeit with correlated outcomes. We show how these methods can be extended to handle the common survey features of stratification, clustering, and sampling weights. We describe how to fit regression models with multiple source reports derived from complex sample surveys using general purpose statistical software. Finally, the methods are illustrated using data from two studies: the Stirling County Study and the Eastern Connecticut Child Survey. PMID- 15344196 TI - Venous hemodynamic changes in the surgical treatment of primary varicose vein of the lower limbs. AB - Venous hemodynamic changes after the surgery of primary varicose veins were evaluated. (Materials and methods) We retrospectively analyzed 1,211 patients (1,407 limbs) who underwent surgery for primary varicose veins from 1994 to 2002. The venous hemodynamics were evaluated using air- plethysmography (APG) preoperatively and one month postoperatively in the viewpoints of ambulatory venous pressure (AVP), venous volume (VV), venous filling index (VFI), and ejection fraction (EF). (Results) The surgical modalities included 958 cases of greater saphenous vein high ligation (GSV HL) and stripping with varicosectomy (VS), 222 cases of short saphenous vein (SSV) HL and VS, 143 cases of external banding valvuloplasty of GSV and VS, and 44 cases using VNUS and VS. The reduction rate of VV was 20.9 +/- 14.1% in the GSV stripping group, 12.0 +/- 14.7% in the GSV valvuloplasty group, 18.3 +/- 16.1% in the VNUS group, and 20.6 +/- 15.9% in the SSV group. The reduction rate of VFI was 63.6 +/- 20.7% in the GSV stripping group, 38.8 +/- 40.9% in the GSV valvuloplasty group, 60.1 +/- 23.9% in the VNUS group, and 37.6 +/- 30.2% in the SSV group. The increasing rate of EF was 25.0 +/- 28.2% in the GSV stripping group, 21.0 +/- 30.0% in the GSV valvuloplasty group, 29.4 +/- 31.9% in the VNUS group, and 30.0 +/- 36.5% in the SSV group. The reduction rate of AVP was 25.4 +/- 32.2% in the GSV stripping group, -6.1 +/- 58.1% in the GSV valvuloplasty group, 28.4 +/- 38.5% in the VNUS group, and 14.1 +/- 49.0% in the SSV group. All of the patients showed improvements in venous hemodynamics by showing a decrease in VV, VFI, AVP, and an increase in EF. However, there was no difference in the change of venous hemodynamics according to the type of surgery. PMID- 15344197 TI - Application of real-time tumor-tracking and gated radiotherapy system for unresectable pancreatic cancer. AB - Herein is reported our experience of radiation therapy using a real-time tumor tracking and gated radiotherapy (RTRT) system for inoperable pancreatic cancer. Three unresectable pancreatic cancer patients were treated with intraoperative electron beam radiation therapy, at the time of open biopsy, and postoperative external beam radiation therapy using an RTRT system with a 2.0 mm diameter gold ball implanted into the pancreas. The total BED'salpha/beta=10 was intended to be equivalent to that of delivering 60 Gy by 2.0 Gy/fraction, while the actual dose schedules were individualized. The movement of the pancreas was analyzed based on the 3-dimensional marker positions during the RTRT. The side effects and tumor responses were evaluated. During the RTRT course, the average movement of markers in the x (left to right), y (cranial to caudal) and z (dorsal to ventral) directions were 3.0 mm (1.7- 5.2 mm), 5.2 mm (3.5 - 6.8 mm) and 3.5 mm (2.7 - 5.1 mm), respectively. During and after the course of postoperative radiation therapy, no acute side effects of RTOG grade II or higher were detected. The objective tumor responses, as evaluated by CT scans 3 months after the treatment, were 2 partial responses and no response in one patient. Using the RTRT technique the margin of treatment planning and the possible errors in target localization were reduced, and the 3-dimensional movement of the internal marker implanted in the pancreas was able to be analyzed. PMID- 15344198 TI - Macrolide resistance in beta-hemolytic streptococci: changes after the implementation of the separation of prescribing and dispensing of medications policy in Korea. AB - This study evaluated the antimicrobial susceptibilities and macrolide resistance mechanisms of beta-hemolytic streptococci (BHS), and an additional objective was to assess the effects of 'the separation of prescribing and dispensing (SPD) of medications' on bacterial resistance rate and distribution of phenotypes and genotypes of erythromycin-resistant BHS by comparing the antimicrobial susceptibility data before (1990- 2000) and after the implementation of SPD at one tertiary care hospital in South Korea. Between the period of January 2001 and December 2002, the minimal inhibitory concentrations of six antimicrobials were determined for 249 clinical isolates of BHS. Resistance mechanisms of erythromycin-resistant (intermediate and resistant) isolates were studied by using the double disk test and PCR. Overall, the resistance rates to tetracycline, erythromycin, and clindamycin were 75.5%, 32.9%, and 32.5%, respectively. Sixty-seven (81.7%) of 82 erythromycin- resistant isolates expressed constitutive resistance to macrolide- lincosamide-streptogramin B antibiotics (a constitutive MLSB phenotype); 11 isolates (13.4%) expressed an M phenotype; and four isolates (4.9%) had an inducible MLSB resistance phenotype. erm(A) was found in isolates with constitutive/ inducible MLSB phenotypes, erm(B) with the constitutive/ inducible MLSB phenotype, and mef(A) with the M phenotype. We found that resistance rates to erythromycin and clindamycin among S. agalactiae, S. pyogenes, and group C streptococci isolates were still high after the implementation of the SPD policy in Korea, and that the constitutive MLSB resistance phenotype was dominant among erythromycin- resistant BHS in this Korean hospital. PMID- 15344199 TI - Increasing prevalence of vancomycin-resistant enterococci, and cefoxitin-, imipenem- and fluoroquinolone-resistant gram-negative bacilli: a KONSAR study in 2002. AB - Continued antimicrobial resistance surveillance can provide valuable information for the empirical selection of antimicrobial agents for patient treatment, and for resistance control. In this 6th annual study for 2002, the susceptibility data at 39 Korean Nationwide Surveillance of Antimicrobial Resistance (KONSAR) hospitals were analyzed. Resistance rates of S. aureus were 67% to oxacillin, and 58% to clindamycin. The ampicillin and vancomycin resistance rates of E. faecium were 89% and 16%, respectively. To penicillin, 71% of S. pneumoniae were nonsusceptible. Resistance rates of E. coli were 11% to cefotaxime, 8% to cefoxitin, and 34% to fluoroquinolone, and those of K. pneumoniae were 22% to ceftazidime, and 16% to cefoxitin. Lowest resistance rates to cephalosporins shown by E. cloacae and S. marcescens were to cefepime, 7% and 17%, respectively. This is the first KONSAR surveillance, which detected imipenem-resistant E. coli and K. pneumoniae. To imipenem, 22% of P. aeruginosa and 9% of Acinetobacter spp. were resistant. Trends of resistances showed a slight reduction in MRSA and in penicillin- nonsusceptible S. pneumoniae, but an increase in ampicillin-resistant E. faecium. Ampicillin-resistant E. coli and H. influenzae remained prevalent. Compared to the previous study, amikacin- and fluoroquinolone- resistant Acinetobacter spp. increased to 60% and 62%, respectively. Ceftazidime- resistant K. pneumoniae decreased slightly, and imipenem- resistant P. aeruginosa and Acinetobacter spp., and vancomycin-resistant E. faecium increased. In conclusion, vancomycin-resistant E. faecium, cefoxitin-resistant E. coli and K. pneumoniae, and imipenem-resistant P. aeruginosa and Acinetobacter spp. increased gradually, and imipenem- resistant E. coli and K. pneumoniae appeared for the first time. Continued surveillance is required to prevent further spread of these serious resistances. PMID- 15344200 TI - Anterior transposition of inferior oblique muscle for treatment of unilateral superior oblique muscle palsy with inferior oblique muscle overaction. AB - Although many weakening procedures for the inferior oblique muscle have been advocated, there is some controversy as to the most beneficial procedure for weakening overacting inferior oblique muscles. This study was undertaken to determine if unilateral anterior transposition of the inferior oblique muscle alone could be a safe and effective procedure for treating unilateral superior oblique palsy from the perspective of hypertropia, inferior oblique overaction, and abnormal head posture. The records of 33 patients, who underwent anterior transposition of the inferior oblique muscle for unilateral superior oblique palsy at our institution between Jan 1995 and Dec 2002, were retrospectively reviewed. The average preoperative inferior oblique overaction was 2.3 +/- 0.64, and the hypertropia in the primary position was 12.3 +/- 7.69 prism diopter (PD). Twenty-six patients showed head tilt to the opposite direction preoperatively. After the anterior transposition of the inferior oblique, inferior oblique overaction was diminished in 32 patients (97%). Twenty-six out of 33 patients (79%) had no hypertropia in the primary position at last postoperative assessment. Of the 26 patients with head tilt before surgery, 21 patients (81%) achieved full correction after surgery. Satisfactory results were obtained in most of the patients in our study with the exception of three patients who required additional surgery. No patient demonstrated postoperative hypotropia in the primary position. None of the patients noticed elevation deficiency or lower lid elevation. The anterior transposition of the inferior oblique was found to be safe and effective for treating superior oblique palsy with secondary overaction of the inferior oblique muscle. PMID- 15344201 TI - Vitreoretinal surgery using transconjunctival sutureless vitrectomy. AB - This report presents the effectiveness of 25-gauge Transconjunctival Sutureless Vitrectomy (TSV) for various vitreoretinal disorders. We performed vitreoretinal surgery on 6 patients using 25-gauge TSV. Minimal or no leakage of intraocular fluid or gas was observed at the entry site. No case required a suture to close the conjunctival or scleral opening site, and no complications resulted from the opening site. Median preoperative visual acuity was 0.04 and median postoperative best corrected visual acuity (BCVA) with a mean follow-up of 12 weeks, was 0.45. Median preoperative intraocular pressure was 12.67 mmHg, and median intraocular pressure on the first postoperative day was 15.67 mmHg. Because transconjunctival sutureless surgery is minimally invasive, it increases the efficiency of vitrectomy, hastens postoperative recovery, and improves outcomes due to the simplified surgical procedure. We feel that the adoption of the 25-gauge TSV would lead to improved patient comfort, care, and management. PMID- 15344202 TI - Surgical outcome of radical maxillectomy in advanced maxillary sinus cancers. AB - We investigated the surgical outcome of radical maxillectomy in advanced maxillary sinus cancers invading through the posterior wall and into the infratemporal fossa. Twenty-eight patients with maxillary sinus squamous cell carcinoma, who visited the Otorhinolaryngology Department at Severance Hospital from March, 1993 to February, 2001 and underwent the surgery, were analyzed retrospectively by reviewing clinical medical records and radiologic test results. The mean follow- up period was 78.8 months.(26-162 months) Local recurrence, sites of local recurrence, and the 2-year disease-free survival rate were analyzed. Of the total 28 cases, 9 cases were T3, and 19 cases were T4. Total maxillectomy was performed in 12 cases (42.9%) and radical maxillectomy in 16 cases (57.1%). Regardless of staging, radical maxillectomy was performed only when cancers invaded through the posterior wall and into the infratemporal fossa. When cancers only maginally or did not invade the posterior wall, total maxillectomy was performed. The 2-year disease-free survival rate was 75% for both total and radical maxillectomy, and the local recurrence rates were 8.3% and 18.7% respectively. All recurrence occurred at the posterior resection margin of the maxillectomy. We strongly recommend the use of radical maxillectomy in the cases of advanced maxillary sinus cancers invading the infratemporal fossa. Radical maxillectomy can provide sufficient safety margins and lower the local recurrence rate. PMID- 15344203 TI - Percutaneous interventional treatment of extracranial vertebral artery stenosis with coronary stents. AB - Stenosis of extracranial vertebral artery (VA) is not an infrequent lesion, and it can pose a significant clinical problem. However, the standard treatment for a significant VA stenosis has still not been established. Here in this study, we report our experiences of VA stenting in 25 patients (age 56.2 +/- 15.2 years, male 76%). The patients had comorbidities as follows: DM (36%), hypertension (64%), Takayasu's (12%) and Behcet's diseases (4%). There were combined involvement of other vessels such as the coronary artery (72%), carotid artery (36%), subclavian artery (32%) and the contralateral vertebral artery (24%). Indications for stenting were prior stroke or symptoms related to vertebrobasilar ischemia in 11 patients, and an asymptomatic but angiographically significant stenosis (> 70% stenosis) in 14 patients. Twenty-three balloon-expandable stents and two self-expandable stents were deployed. A drug-eluting coronary stent and distal balloon protection device were each used in one case. A technically successful procedure was achieved in all patients. The baseline reference diameter was 4.7 +/- 1.3 mm, minimal luminal diameter (MLD) 1.0 +/- 0.6 mm (diameter stenosis 77.8 +/- 12.5%) and lesion length 6.4 +/- 3.9 mm. After stenting and adjuvant dilation, the MLD was increased to 4.5 +/- 0.9 mm (diameter stenosis 3.1 +/- 17.9%). There were no procedure-related complications. During the further follow-up period of 25 (3-49) months, no stroke or death occurred. Restenosis was observed in 4 (30.8%) of 13 eligible patients. In conclusion, VA stenting is feasible with a high degree of technical success, and this treatment is associated with a relatively low incidence of procedure-related complications. However, a relative high rate of in-stent restenosis remains as a problem to be resolved. PMID- 15344204 TI - The impact of the preoperative severity of target-vessel stenosis on the short term patency of radial artery grafts. AB - The fate of a grafted radial artery remains unknown. The purpose of this study was to determine whether the preoperative severity of stenosis of the target vessel influence short-term patency of radial artery (RA) grafts used as coronary artery bypass conduits. In 54 patients who had coronary artery bypass grafting (CABG) with RA grafts, RA patency was determined with multi-slice computed tomography (MSCT) 1 year after CABG. These patients were divided into three groups on the basis of the percentage of the target vessel stenosis: mild (< 60%, n=17), moderate (60% to 79%, n=19), and severe (>/= 80%, n=18). MSCT was also performed 1 week later to exclude early occlusion of RA grafts. In 3 patients, the MSCT failed to adequately discriminate the status of the RA graft due to poor image resolution. The overall incidence of RA occlusion was 23.5% (12 of 51) at 1 year in the entire population. The mild stenosis, moderate stenosis and severe stenosis group showed an occlusion rate of 50% (8 of 16), 23.5% (4 of 17) and 0% (0 of 18), respectively. The severe stenosis group had significantly lower rate of RA graft occlusion compared to the mild stenosis group (p < 0.001) and moderate stenosis group (p < 0.05). No difference in occlusion between grafts used for the different coronary artery branches could be demonstrated. Preoperative severity of the target coronary artery significantly affected the short-term RA grafts patency. Correct indication is the key factor for short-term RA patency. PMID- 15344205 TI - Change in pulmonary function following empyemectomy and decortication in tuberculous and non-tuberculous chronic empyema thoracis. AB - Chronic empyema thoracis results from various etiologies. Improvement in pulmonary function after empyemectomy and decortication has proved difficult to predict when the etiology of chronic empyema thoracis is tuberculosis. The purpose of this study was to confirm the changes in pulmonary function according to the etiology after an operation. Sixty-five patients were classified into two groups according to their etiology: Group A (tuberculous) and Group B (non tuberculous), and they were retrospectively evaluated with regard to their forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), percentage of predicted normal value of FEV1 (% FEV1) and FVC (%FVC). Empyemectomy and decortication was performed for all the patients and the two groups were similar in age, gender and preoperative spirometric parameters. In Group A (n=41), the pre- and postoperative mean values were 2.31 L and 2.8 L in FEV1, 65.8% and 80.5% in %FEV1, 2.62 L and 2.55 L in VC, 61.9% and 71.8% in %VC, respectively. In Group B (n=24), the pre- and postoperative mean values were 2.13 L and 2.49 L in FEV1, 66.4% and 73.8% in %FEV1, 2.55 L and 2.95 L in FVC, 64.9% and 71.8% in %FVC, respectively. All the spirometric parameters improved significantly in both groups compared to their preoperative values. However, no significance was shown in the rate of increase of the spirometric parameters between the two groups. In conclusion, improvement of lung function is expected after empyemectomy and decortication, regardless of the etiology of the chronic empyema thoracis. PMID- 15344206 TI - Characteristics of potassium and calcium currents of hepatic stellate cells (ito) in rat. AB - Hepatic stellate cells (HSCs) are known to play a role in the pathogenesis of the increased intrahepatic vascular resistance found in chronic liver diseases. The aim of this study was to evaluate the K+ and Ca2+ currents in cultured HSCs from rat liver, through the patch-clamp technique. Most cells were positive for desmin immunostain after isolation and in alpha-smooth muscle actin immunostain after 10 - 14 days of culturing. Outward and inward rectifying K+ currents were confirmed. Two different types of K+ currents were distinguished: one with the inward rectifying current and the other without. The outward K+ currents consisted of at least four components: tetraethylammonium (TEA)-sensitive current, 4 aminopyridine (4-AP)-sensitive current, pimozide-sensitive current and three blocker-resistant current. The peaks of the outward K+ currents evoked by a depolarizing pulse were decreased to 32.0 +/- 3.0, 62.8 +/- 3.7 and 32.8 +/- 3.5% by 5 mM TEA, 2 mM 4-AP and 15 micro M pimozide, respectively. Moreover, the combined application of three blockers caused 86.6 +/- 4.8% suppression. The inward currents evoked hyperpolarizing pulses were inwardly rectifying and almost blocked by Ba2+. Elevation of external K+ increased the inward current amplitude and positively shifted its reversal potential. Voltage- dependent Ca2+ currents which were completely abolished by Cd2+ and nimodipine were detected in 14 day cultured HSCs. In this study, the cultured HSCs were found to express outward K+ currents composed of multiple pharmacological components, Ba2+-sensitive inward rectifying K+ current and L-type Ca2+ current. PMID- 15344207 TI - The diagnostic value of the adenosine deaminase activity in the pleural fluid of renal transplant patients with tuberculous pleural effusion. AB - The assessment of the adenosine deaminase activity (ADA) in the pleural effusion is used for the diagnosis of tuberculous pleural effusion (TPE). To examine whether the procedure can be applied to immunocompromised patients, we analyzed the ADA activity in the pleural fluid of renal transplant recipients. We studied 23 renal transplant patients with TPE (21 men and 2 women; the mean age, 33 years). They were treated at the Yonsei University Hospital between January 1985 and December 2001. Patients with granuloma in the pleural biopsy specimen or positive for Mycobacterium tuberculosis in the pleural fluid culture were recruited. The ADA activity in the pleural effusion of 23 renal transplant patients with TPE was compared with 23 immunocompetent patients with TPE. The mean ADA activity was 69.5 +/- 4.6 in renal transplant patients and 65.0 +/- 4.9 U/L in immunocompetent patients. Applying the 40 U/L cut-off point, the positivity of ADA was 91.3% in renal transplant patients, and 86.9% in immunocompetent patients. We thus concluded that the measurement of ADA in the pleural fluid is a useful means in the diagnosis of TPE in renal transplant patients. PMID- 15344208 TI - Comparison of outcomes between Burch colposuspension with and without concomitant abdominal hysterectomy. AB - A total abdominal hysterectomy may cause a postoperative vesicourethral dysfunction due to an injury to the pelvic nerves. However, many incontinent women with benign diseases of the uterus and its adnexae have undergone a Burch colposuspension with a concomitant abdominal hysterectomy. This study was undertaken to compare the outcomes of a Burch colposuspension performed alone with that of a Burch with a concomitant abdominal hysterectomy. This study included 132 women, who, were treated for primary urinary incontinence from February 1999 to February 2002 and were diagnosed with stress urinary incontinence by means of the urodynamic test at the Department of Obstetrics and Gynecology at Yonsei University Hospital. Forty-two women underwent a Burch colposuspension alone (Burch group) and 90 women underwent a Burch colposuspension with a concomitant abdominal hysterectomy (hysterectomy group). Between the Burch and hysterectomy groups, the mean age, parity, menopausal rate, Hormone Replacement Therapy (HRT) rate, 1 year follow-up outcomes and postoperative complications were compared using the subjective and objective stress tests according to the retrospective chart review. The mean age (54.6 +/- 0.5 vs 58.6 +/- 9.2 years, p=0.382), parity (3.3 +/- 1.2 vs 3.6 +/- 1.7), menopausal rate (71.4 vs 77.7%), or HRT rate (23.3 vs 11.2%) of the two groups were similar. Complications related to surgery were encountered in 5 patients (11.9%) in the Burch group and in 7 patients (7.8%) in the hysterectomy group (p=0.842). One year follow-up subjective symptoms were encountered in 2 patients in the Burch group and in 4 patients in the hysterectomy group (p=1.00). The stress test was positive in only one patient in the hysterectomy (p=1.00). No significant difference was observed in the 1 year follow-up outcomes, which were 91.4% (32/35 patients) in the Burch and 91.2% (73/80) in the hysterectomy groups. The results showed that there were no adverse effects on the 1 year follow-up outcomes or complications in patients who underwent a Burch colposuspension with an abdominal hysterectomy. PMID- 15344209 TI - Can a 12 core prostate biopsy increase the detection rate of prostate cancer versus 6 core?: a prospective randomized study in Korea. AB - Several studies suggest that standard 6 core sextant transrectal ultrasound (TRUS) guided biopsies of the prostate provides insufficient material to adequately detect clinically important prostate cancer, and that a larger biopsy cores may improve the cancer detection rate. We performed a prospective randomized trial by comparing 6 and 12 core prostate biopsies to determine whether doubling the number of cores in a sextant biopsy improves the prostate cancer detection rate. We randomized 240 men with an elevated serum total prostate specific antigen (PSA) level, abnormal digital rectal examination (DRE) and/or TRUS suspicious for prostate cancer into a 6 core biopsy group and 12 core biopsy group from Jan. 2002 to Jan. 2003. We acquired 3 cores from the right and left prostate lobes for the 6 core biopsy group and three additional cores from each side more peripheral than a 6 core for the 12 core biopsy group. The 6 core and 12 core biopsy groups were well matched with no significant differences in age, prostate volume, PSA and PSA density. The overall cancer detection rate by prostate biopsy was 15.8% (38/240) and the cancer detection rate was not significantly different between the 6 core biopsy group (14.4%, 17/118) and 12 core biopsy group (17.2%, 21/122) (p=0.60). Our study demonstrates no statistically significant improvement in prostate cancer detection rate by increasing the number of biopsy cores. In conclusion, we believe that the standard 6 core sextant biopsy of the prostate is as effective at detecting prostate cancer as a 12 core biopsy in Korean men. PMID- 15344210 TI - Determinants of one-year response of lumbar bone mineral density to alendronate treatment in elderly Japanese women with osteoporosis. AB - The purpose of this study was to determine factors that could predict the one year response of the lumbar bone mineral density (BMD) to alendronate treatment in elderly Japanese women with osteoporosis. Eighty-five postmenopausal women with osteoporosis, all of whom were between 55-88 years of age, were treated with alendronate (5 mg daily) for 12 months. Serum calcium, phosphorus, and alkaline phosphatase (ALP) and urinary NTX levels were measured at the baseline and 6 months, and lumbar (L1-L4) BMD was measured by dual energy X-ray absorptiometry at the baseline and 12 months. Multiple regression analysis was used to determine factors that were correlated with the percent change in lumbar BMD at 12 months. Lumbar BMD increased by 8.1 % at 12 months with a reduction in the urinary NTX level by 51.0 % at 6 months. Baseline lumbar BMD (R2=0.226, p < 0.0001) and percent changes in serum ALP and urinary NTX levels (R2=0.044, p < 0.05 and R2=0.103, p < 0.001, respectively) had a negative correlation with the percent change in lumbar BMD at month 12, while the baseline number of prevalent vertebral fractures (R2=0.163, p < 0.001), serum ALP level, and urinary NTX level (R2=0.074, p < 0.05 and R2=0.160, p < 0.001, respectively) had a positive correlation with it. However, baseline age, height, body weight, body mass index, years since menopause, serum calcium and phosphorus levels, and percent changes in serum calcium and phosphorus levels at 6 months did not have any significant correlation with the percent change in lumbar BMD at 12 months. These results suggest that lumbar BMD was more responsive to one-year of alendronate treatment in elderly osteoporotic Japanese women with lower lumbar BMD, more prevalent vertebral fractures, and higher bone turnover, who showed a greater decrease in bone turnover at 6 months, regardless of age, years since menopause, and physique. Alendronate may be efficacious in elderly Japanese women with evident osteoporosis that is associated with high bone turnover, and the percent changes in serum ALP and urinary NTX levels at 6 months could predict the one-year response of lumbar BMD to alendronate treatment. PMID- 15344211 TI - Glutathione S-transferase M1 status and breast cancer risk: a meta-analysis. AB - It is not yet clear whether Glutathione S-transferase M1 (GSTM1) polymorphisms affect the risk of breast cancer. The aim of this study is to provide a comprehensive meta-analysis of all the available, published case-control studies on the extent of the possible association between GSTM1 polymorphisms and susceptibility to breast cancer. Twenty case-control studies on GSTM1 and breast cancer were identified using both PUBMED and a manual search. Meta-analysis was conducted by the Peto method. Subgroup analyses were undertaken, in order to explore the relationship between effect sizes and the study characteristics. The overall odds ratio (OR) was found to be 1.06 (95% CI, 0.99-1.14). The OR for post menopausal women with GSTM1 deficiency was determined to be 1.19 (95% CI, 1.05 1.34). In populations with a low frequency of GSTM1 deficiency, a greater increase was observed (OR, 1.20; 95% CI, 1.08-1.34). Furthermore, the highest associations were found in post-menopausal women with a low frequency of GSTM1 deficiency (OR, 1.44; 95% CI, 1.20-1.73). The fact that GSTM1 deficiency is not rare in the general population implies that the attributable risk for breast cancer could be sizable. Further studies focusing on the structure of haplotype blocks of GSTM1 are required in order to find a specific haplotype with a predisposing breast cancer susceptibility allele. PMID- 15344212 TI - Mechanics and fatigability of the rat soleus muscle during early reloading. AB - In order to elucidate muscle functional changes by acute reloading, contractile and fatigue properties of the rat soleus muscle were investigated at three weeks of hindlimb suspension and the following 1 hr, 5 hr, 1 d, and 2 weeks of reloading. Compared to age-matched controls, three weeks of unloading caused significant changes in myofibrillar alignments, muscle mass relative to body mass (-43%), normalized tension (-35%), shortening velocity (+143%), and response times. Further significant changes were not observed during early reloading, because the transitional reverse process was gradual rather than abrupt. Although most of the muscle properties returned to the control level after two weeks of reloading, full recovery of the tissue would require more than the two-week period. Delayed recovery due to factors such as myofibrillar arrangement and fatigue resistance was apparent, which should be considered for rehabilitation after a long-term spaceflight or bed-rest. PMID- 15344213 TI - Effects of vitamin E supplementation on oxidative stress in streptozotocin induced diabetic rats: investigation of liver and plasma. AB - This experimental study was designed to investigate the effects of vitamin E supplementation, especially on lipid peroxidation and antioxidant status elements 3/4 namely, glutathione (GSH), CuZn superoxide dismutase (CuZn SOD), and glutathione peroxidase (GSH Px), both in blood and liver tissues of streptozotocin (STZ) diabetic rats. The extent to which blood can be used to reflect the oxidative stress of the liver is also investigated. In diabetic rats, plasma lipid peroxide values were not significantly different,from control,whereas erythrocyte CuZn SOD (p < 0.01), GSH Px (p < 0.001) activities and plasma vitamin E levels (p < 0.001), were significantly more elevated than controls. Vitamin E supplementation caused significant decreases of erythrocyte GSH level (p < 0.01) in control rats and of erythrocyte GSH Px activity (p < 0.05) in diabetic rats. Liver findings revealed significantly higher lipid peroxide (p < 0.001) and vitamin E (p < 0.01) levels and lower GSH (p < 0.001), CuZn SOD (p < 0.001) and GSH Px (p < 0.01) levels in diabetic rats. A decreased hepatic lipid peroxide level (p < 0.01) and increased vitamin E/lipid peroxide ratio (p < 0.001) were observed in vitamin E supplemented, diabetic rats. A vitamin E supplementation level which did not cause any increase in the concentration of the vitamin in the liver or blood, was sufficient to lower lipid peroxidation in the liver. Vitamin E/lipid peroxide ratio is suggested as an appropriate index to evaluate the efficiency of vitamin E activity,independent of tissue lipid values. Further, the antioxidant components GSH, GSH Px and CuZn SOD and the relationships among them, were affected differently in the liver and blood by diabetes or vitamin E supplementation. PMID- 15344214 TI - Temperature changes in superficial and deep tissue layers with respect to time of cold gel pack application in dogs. AB - Despite the widespread clinical use of cryotherapy, there is only limited and inconsistent data on application times. The aim of this study was to determine the changes in tissue temperature and the duration of this effect. In this experimental study, five adult dogs were used. A cold gel pack (10 x 20 cm) was applied transversally over the right leg femoral region. Temperatures were recorded simultaneously: rectal by a mercury thermometer; right leg skin by probe of Nihon Kohden 6000 polygraph; and right leg subcutaneous, intramuscular, and periosteal, and left leg intramuscular temperatures by a fluorooptic biomedical fiber optic (0.6 mm diameter) thermometer connected to a computer system. Total system accuracy was 0.01 degrees C. Cold gel packs were applied for 10, 15, 20, 25, and 30 minutes duration. The results can be summarized as cooling and rewarming data. 1) The superficial tissues such as skin and subcutaneous demonstrated the most rapid and profound cooling effect. The deeper tissues such as bone and muscle exhibited a smaller and more gradual decline in temperature. 2) There was a prolonged rewarming period in all tissues after the removal of the cold gel pack but this period was longer in deeper tissues. According to cold gel pack application time, the rewarming time in intramuscular layers to baseline or plateau temperatures was about: 60 +/- 3 minutes for 10 minutes application, 100 +/- 4 for 15, 130 +/- 5 for 20, 140 +/- 7 for 25, and 145 +/- 8 for 30. It can be concluded from these results that with increased cold gel pack application time, deep tissue temperature decreased and the duration of cooling effect increased. However, the data indicated that the length of application time and the duration of cooling effect were not linearly related. Especially after 20 minutes of application this ratio decreased progressively. There may be implications of these results for clinical practice. PMID- 15344215 TI - A case of medullomyoblastoma of cerebellopontine angle mimicking acoustic neuroma. AB - Medulloblastoma is a common malignant central nervous system neoplasm found mainly in children. One the contrary, medulloblastoma of the cerebellopontine angle, the location of the tumor is very unusual. This is the the first case of the medullomyoblastoma, a rare form of medulloblastoma, occurring in the cerebellopontine angle. A 15-year-old boy experienced a sudden hearing loss in the left ear. Conservative medical treatment failed, and temporal MR imaging revealed a heterogeneously enhancing mass at the left cerebellopontine angle cistern and in the internal auditory canal; therefore, the lesion was regarded as a typical acoustic neuroma. Few days before surgery, an ipsilateral facial palsy developed, and a follow-up MR imaging showed a rapid growth of the previous lesion. The extended translabyrinthine approach permitted surgical removal. And under pathological diagnosis of malignancy, radiation therapy and series of chemotherapy was performed. PMID- 15344216 TI - Burkitt's lymphoma representing periportal infiltrating mass on CT. AB - Imaging findings of secondary hepatic lymphoma have been reported as variable, ranging from single or multiple small nodules to diffuse infiltrative tumor patterns. We hear present a rare case report concerning aggressive B cells, secondary Burkitt's lymphoma in non-AIDS demonstrating a surprising periportal lymphoma infiltration, without upper abdominal lymphadenopathy or splenomegaly on the sonography and CT scans. Clinically, the case was characterized by atypical and highly aggressive course, with the patient presenting an abruptly developed obstructive jaundice with rapidly deteriorating hepatic function that could be indicative of cholestatic hepatitis, which differs in its clinical manifestations from hepatic lymphoma without functional deterioration in respect of its non tissue destructive growth pattern. We suggest that hepatic lymphoma can sometimes be consistent with periportal infiltrating homogeneous mass, with no lymphadenopathy or splenomegaly on the imaging examination, with a predictable aggressive clinical course of the disease and poor prognosis. PMID- 15344217 TI - Kearns-Sayre syndrome -3 case reports and review of clinical feature. AB - Kearns-Sayre syndrome, first described by Kearns and Sayre in 1958, is a rare disorder consisting of ptosis, limited movement of both eyes and atypical retinal pigmentary change (salt-pepper like appearance). Most cases have shown an increase in the concentration of mitochondria and ragged-red fiber under Gomori trichrome staining on muscle biopsy. Occasionally, it is combined with other neurologic and endocrinologic symptoms such as ataxia, dementia, diabetes, and hyperaldosteronism. We recently experienced three cases of male teenaged patients who expressed the clinical features of Kearns-Sayre syndrome. PMID- 15344218 TI - Malignant fibrous histiocytoma (MFH) in axilla. AB - Palpable axilla mass in woman is relatively rare. Almost all palpable lumps in axilla are axillarys accessory breasts without mass lesion. All diseases develop in breast can also develop in axillarys accessory breasts and other soft tissue mass can occur in axilla. Malignant fibrous histiocytoma (MFH) is the most common malignant soft tissue tumor, but axillarys MFH is extremely rare. We report our experience with a 75-year-old woman with MFH in axilla, treated with wide excision. PMID- 15344219 TI - CT findings of surgically verified acute invasive small bowel anisakiasis resulting in small bowel obstruction. AB - Acute invasive small bowel anisakiasis is an extremely rare cause of small bowel obstruction. The authors report a case of surgically verified small bowel anisakiasis resulting in small bowel obstruction. A 54-year-old man presented with suddenly developed diffuse abdominal pain after ingestion of raw fish. The peripheral blood examination showed leukocytosis without eosinophilia. CT showed a long segment of thickened small bowel accompanied by a focal narrowed portion and combined with ascites. When these findings are noted in patients with a history of recent ingestion of raw or undercooked fish, the diagnosis of small bowel anisakiasis should be considered in order to avoid application of unnecessary surgical treatment, in spite of the severity of the abdominal pain and bowel obstruction. PMID- 15344220 TI - The effect of selective tibial neurotomy and rehabilitation in a quadriplegic patient with ankle spasticity following traumatic brain injury. AB - Ankle spasticity following brain injury leads to abnormal posture and joint contracture; making standing or walking impossible. This study investigates the efficacy of selective tibial neurotomy (STN) and intensive rehabilitation in a patient who suffered ankle spasticity after brain injury. This case describes a 37-year-old man whose traumatic brain injury (TBI) resulted in severe right ankle spasticity and contracture. He was unable to stand due to severe right ankle spasticity and contracture. Intensive rehabilitation and STN allowed him to walk without brace at 6 months and run at 12 months after STN. STN is an effective procedure to resolve localized spasticity of the ankle and it may be considered as a management strategy after local injection to alleviate ankle spasticity and/or contracture prior to orthopaedic surgery. PMID- 15344221 TI - Management of detachment of pilot balloon during intraoral repositioning of the submental endotracheal tube. AB - Submental endotracheal intubation for surgery was used as an alternative to nasotracheal intubation in patients with craniomaxillofacial injury. Generally extubation was performed in the operation room by pulling the tube through the submental incision site. When extubation is not indicated, intraoral indwelling is preferred to submental intubation. We report a case of a 35-year-old male patient with multiple facial bone fractures. At the end of the surgery, we noticed the oropharyngeal edema, and so the submental intubation was converted into a standard orotracheal intubation. During that procedure, the pilot balloon was accidentally detached from the endotracheal tube. The situation was managed by cutting a pilot tube from a new, unused endotracheal tube and connecting it to the intubated tube using a needle connector. PMID- 15344222 TI - Involvement of MAP kinases in apoptosis of macrophage treated with Trichomonas vaginalis. AB - A primitive protozoan parasite Trichomonas vaginalis selectively activates the signal transduction pathways in macrophages (RAW264.7). This study evaluated the correlation of these signaling pathways and T. vaginalis-induced cell apoptosis. In macrophages infected with T. vaginalis, apoptosis was assessed on the basis of DNA fragmentation on agarose gel electrophoresis. Infection of macrophages with T. vaginalis induced tyrosine phosphorylation of several proteins. Infected cells with T. vaginalis were shown to associate with phosphorylation of the extracellular signal-regulated (ERK)1/2 kinase, p38, c-Jun N-terminal kinase (JNK) mitogen-activated protein (MAP) kinases on Western blot analysis. The present finding also demonstrated a link between the ERK1/2, JNK and p38 apoptotic pathways that was modulated by T. vaginalis infection. PMID- 15344223 TI - Peptide mimotopes of Neisseria meningitidis group B capsular polysaccharide. AB - The antigenic similarity between Neisseria meningitidis group B (NMGB) capsular polysaccharide (PS) and human polysialic acid (PSA) has hampered the development of a NMGB PS-based vaccine. But the possibility of a safe vaccine based on NMGB PS has been demonstrated by the existence of the NMGB PS-associated nonautoreactive epitope, which is distinct from those present on human PSA. To obtain peptide mimotopes of NMGB PS, we used HmenB3, a protective and nonautoreactive monoclonal antibody, to screen a phage library with 12 amino acids. We obtained 23 phage clones that bound to HmenB3 but not in the presence of E. coli K1 PS [which is alpha(2-8)-linked PSA like NMGB PS]. The clones contained 3 mimotopes and differed from previously described NMGB PS mimotopes. Immunization with a synthetic peptide of one mimotope elicited anti-NMGB antibodies in BALB/c mice. These mimotopes may be useful in the development of group B meningococcal vaccines. PMID- 15344224 TI - Is rhabdomyolysis a rare manifestation in celiac disease? PMID- 15344225 TI - Proceedings of the Workshop on Receptor-Binding Radiotracers 2003. PMID- 15344226 TI - Pathologic quiz case. An 18-year-old man with pain and swelling of the left hand. Chondroblastoma. PMID- 15344227 TI - Zero-Check: a zero-knowledge protocol for reconciling patient identities across institutions. PMID- 15344229 TI - Proceedings of the 7th International Symposium on Mutations in the Human Genome. 2-6 July 2003, Cairns, Australia. PMID- 15344228 TI - Defective repair replication of DNA in xeroderma pigmentosum. 1968. AB - Most forms of the human hereditary disease xeroderma pigmentation (XP) are due to a defect in nucleotide excision repair of DNA damage in skin cells associated with exposure to sunlight. This discovery by James Cleaver had an important impact on our understanding of nucleotide excision repair in mammals. PMID- 15344230 TI - Mood disorders: an overview--Part II. PMID- 15344231 TI - Darwinism and psychiatry. PMID- 15344232 TI - PTSD and depression. PMID- 15344233 TI - Narcotics prescription in Switzerland. PMID- 15344234 TI - Mood disorders: an overview--Part III. PMID- 15344235 TI - A view of hypnosis. PMID- 15344236 TI - Inheriting dementia. PMID- 15344237 TI - New light on CFS. PMID- 15344238 TI - How should attention deficit disorder be described? PMID- 15344239 TI - Cancer and the mind. PMID- 15344240 TI - Alcoholism in women. PMID- 15344241 TI - A gene for neophilia? PMID- 15344242 TI - What do psychiatrists need to know about prion diseases? PMID- 15344243 TI - Alternative medicine. Time for a second opinion. PMID- 15344244 TI - Listening to St. John's wort. PMID- 15344245 TI - Sleep apnea. No rest for the weary without proper diagnosis. PMID- 15344246 TI - Prevention. Resisting antibiotic use. PMID- 15344247 TI - Medical progress. On the trail of new arthritis treatments. PMID- 15344248 TI - By the way, doctor. I've always used tap water for drinking and cooking, but I wonder if it's safe. Would I be better off buying a water purifier? PMID- 15344249 TI - Monitoring blood pressure reduces need for drugs. PMID- 15344250 TI - Mammogram timing. PMID- 15344251 TI - Prevention. Fighting cancer with food. PMID- 15344253 TI - Herbal remedies. Ginkgo biloba may slow dementia. PMID- 15344252 TI - Heart disease. Keeping arteries open after angioplasty. PMID- 15344254 TI - By the way, doctor. "I've heard that over-the-counter laxatives cause cancer. Is this true"? PMID- 15344255 TI - Controlled drugs. Broadening the use of pain medications. PMID- 15344256 TI - Handle common pain reliever with care. PMID- 15344257 TI - Occupational therapy keeps older people healthy. PMID- 15344258 TI - Cholesterol and stroke. PMID- 15344259 TI - Re: Evidence for an "epidemic" of myopia. PMID- 15344261 TI - Galactagogue? Galactogogue? PMID- 15344260 TI - Response to research spotlight published in February 2004 issue. PMID- 15344262 TI - Chemical analysis and cellular imaging with quantum dots. AB - Quantum dots are tiny light-emitting particles on the nanometer scale. They are emerging as a new class of biological labels with properties and applications that are not available with traditional organic dyes and fluorescent proteins. Their novel properties such as improved brightness, resistance against photobleaching, and multicolor light emission, have opened new possibilities for ultrasensitive chemical analysis and cellular imaging. In this Research Highlight article , we discuss the unique optical properties of semiconductor quantum dots, surface chemistry and bioconjugation, current applications in bioanalytical chemistry and cell biology, and future research directions. PMID- 15344263 TI - Rapid response nursing. PMID- 15344264 TI - Increasing diversity at the top of the NHS. Interview by Mahua Chatterjee. PMID- 15344265 TI - [What examinations are necessary in an exposed, asymptomatic child with a positive tuberculin skin test and normal chest x-ray?]. PMID- 15344266 TI - [Under what circumstances and how should latent tuberculosis be treated?]. PMID- 15344267 TI - [What are the new diagnostic tools for tuberculosis? What is their role in disease management and what are their indications?]. PMID- 15344268 TI - [What are the new diagnostic tools in tuberculosis? What is their role in disease management and what are their indications? Gene amplification tests]. PMID- 15344269 TI - [What attitude to adopt before the sequellae of tuberculosis are evident on chest x-ray? Before a chest x-ray image, how do you confirm that a disturbance is a tuberculosis sequellae?]. PMID- 15344270 TI - [What are the new diagnostic tools in tuberculosis? What is their role in disease management and what are their indications? Diagnostic role of adenosine deaminase activity measurement in tuberculosis]. PMID- 15344271 TI - [What are the new diagnostic tools in tuberculosis? What is their role in disease management and what are their indications? Diagnostic role of B- and T-cell response evidence in the course of tuberculosis]. PMID- 15344272 TI - [What are the new diagnostic tools in tuberculosis? What is their role in disease management and what are their indications? Diagnostic tools of the future]. PMID- 15344273 TI - [What attitude to adopt before the sequellae of tuberculosis are evident on chest x-ray? Risk of progression and role of chemoprophylaxis before the images of inactive tuberculosis depending on prior treatment and course]. PMID- 15344274 TI - [When should standard treatment not be applied for the tuberculosis patient?]. PMID- 15344275 TI - [Can the medical-social and administrative organization improve the individual and collective management of tuberculosis in France?]. PMID- 15344276 TI - Abstracts of the Serono Symposia International, 2nd International Meeting on Multiple System Atrophy. Rome, Italy, June 17-18, 2004. PMID- 15344277 TI - The rising utilization and costs of prescription drugs. PMID- 15344278 TI - 9th International Conference on AD and related disorders (ICAD). PMID- 15344279 TI - Aggressive management of stroke recommended. PMID- 15344282 TI - The growth in diagnostic imaging utilization. PMID- 15344285 TI - Proceedings of the 3rd International Conference on Oxygen/Nitrogen Radicals: Cell Injury and Disease. Morgantown, West Virginia, USA, 1-5 June 2002. PMID- 15344289 TI - New recommendation for childhood influenza vaccine. PMID- 15344290 TI - Medical report cards and hospital quality. PMID- 15344291 TI - The effect of ADHD on educational outcomes. PMID- 15344292 TI - An unusual case of scalp laxity following weight reduction. AB - The advent of fast foods, increased mechanization,advanced telecommunication and improved living standards have caused human beings to adopt sedentary life styles leading to lesser means of expending calories, accumulation of body fat and tendency to be obese. Plastic surgery can be of great help in supporting patients in their attempts at weight reduction by removing resistant body fat and excising redundant skin that may not succumb to other means of weight reduction. The authors present an unusual case of a young gentleman who developed a rare laxity of the scalp following a drastic weight reduction. Scalp reduction led to a significant cosmetic improvement and psychological acceptance of body image. A review of literature and discussion on the management of this unusual case follows. PMID- 15344293 TI - Pasteur in his laboratory. PMID- 15344294 TI - MDCT appearance of idiopathic saccular aneurysm of the inferior vena cava. PMID- 15344295 TI - Collision adrenal tumors on PET/CT. PMID- 15344296 TI - Severe hypercapnia mimicking hypoxic brain edema on cerebral CT. PMID- 15344297 TI - Fat necrosis in a chest wall lipoma. PMID- 15344298 TI - A 14-3-3 protein of Chlamydomonas reinhardtii associated with the endoplasmic reticulum: nucleotide sequence of the cDNA and the corresponding gene and derived amino acid sequence. AB - Two major 14-3-3 proteins of the unicellular green alga Chlamydomonas reinhardtii were purified and partially sequenced. The obtained data show that the 30-kDa isoform predominant in the cytosol is encoded by a previously cloned and sequenced 14-3-3 cDNA whereas the 27-kDa isoform represents a new 14-3-3 protein which is largely associated with the endoplasmic reticulum (ER). Therefore, the corresponding cDNA was cloned and sequenced. The nucleotide sequence of this new cDNA species and the derived amino acid sequence differ considerably from the previously cloned Chlamydomonas 14-3-3 cDNA. The conclusion that the divergent evolution of the corresponding genes must have started rather early as compared to the 14-3-3 genes of other organisms was corroborated by their different genomic organization. The amino acid sequences of both 14-3-3 isoforms were comparatively analysed to find differences which might be responsible for their differential binding to the ER. PMID- 15344299 TI - 12 surefire ways to lose patients. PMID- 15344301 TI - The impact of health economics on healthcare delivery. A primary care perspective. AB - With the increasing emphasis on providing efficient and equitable services from primary care and against a background of increasing demands on limited resources, economic theory seeks to facilitate both the direction of primary care and the decisions that are made within it. This paper argues that the impact of health economics, particularly at the microeconomic level, has been limited. This is because health economists have failed to recognise the importance of context, and also reflects their attempts to force reality into a disciplinary matrix which is not always accessible and acceptable to end users. Argument is made for a closer relationship between health economists and those who commission and deliver primary care. It is also desirable to develop pragmatic decision-making frameworks which draw upon economic concepts and principles but reflect the realities of the environment in which they are applied. PMID- 15344302 TI - Cost effectiveness of therapies for atrial fibrillation. A review. AB - Atrial fibrillation is the most common supraventricular tachyarrhythmia encountered in clinical practice, affecting over 5% of persons over the age of 65 years. A common pathophysiological mechanism for arrhythmia development is atrial distention and fibrosis induced by hypertension, coronary artery disease or ventricular dysfunction. Less frequently, atrial fibrillation is caused by mitral stenosis or other provocative factors such as thyrotoxicosis, pericarditis or alcohol intoxication. Depending on the extent of associated cardiovascular disease, atrial fibrillation may produce haemodynamic compromise, or symptoms such as palpitations, fatigue, chest pain or dyspnoea. Arrhythmia-induced atrial stasis can precipitate clot formation and the potential for subsequent thromboembolism. Comprehensive management of atrial fibrillation requires a multifaceted approach directed at controlling symptoms, protecting the patient from ischaemic stroke or peripheral embolism and possible conversion to or maintenance of sinus rhythm. Numerous randomised trials have demonstrated the efficacy of warfarin--and less so aspirin (acetylsalicylic acid)--in reducing the risk of embolic events. Furthermore, therapeutic strategies exist that can favourably modify symptoms by restoring and maintaining sinus rhythm with cardioversion and antiarrhythmic prophylaxis. However, the risks and benefits of various treatments is highly dependent on patient-specific features, emphasising the need for an individualised approach. This article reviews the findings of cost-effectiveness studies published over the past decade that have evaluated different components of treatment strategies for atrial fibrillation. These studies demonstrate the economic attractiveness of acute management options, long term warfarin prophylaxis, telemetry-guided initiation of antiarrhythmic therapy, approaches to restore and maintain sinus rhythm, and the potential role of transoesophageal echocardiographic screening for atrial thrombus prior to pharmacological or electrical cardioversion. Further, we discuss the merits and limitations of the cost-effectiveness analyses in the context of overall treatment strategies. Finally, we identify areas that will require additional research to achieve the goal of effective and economically efficient management of atrial fibrillation. PMID- 15344303 TI - Socioeconomic issues affecting the treatment of obesity in the new millennium. AB - The prevalence of obesity among the populations of most developed countries has increased to such an extent that the healthcare and social security/disability system will accumulate direct and indirect costs related to obesity that will be more substantial than those for any other primary disease within this generation. For the past decade, the Healthcare Financing Agency, which oversees the Medicare and Medicaid programmes, has required all physicians and healthcare agencies serving beneficiaries of these programmes to include diagnoses using codes established by the ninth revision of the World Health Organization's International Classification of Diseases. This coding system actually distorts data collection and undermines appropriate medical insurance reimbursement for the treatment of obesity. Societal prejudices, inability of governmental agencies to address future concerns and the business community's attempts to control healthcare costs without addressing the underlying issues contributing to these costs have led to confusion on how to confront this emerging epidemic. How will we develop the scientific knowledge and the political willpower to confront this epidemic? First, we need more accurate methods for classifying obesity and for measuring the cost of treatment. We can then determine if it is more cost effective to prevent or treat obesity early in its evolution or pay for its consequences in the form of treatment costs associated with its multiple comorbid diseases, such as hypertension, other cardiovascular disorders, diabetes mellitus, osteoarthritis and cancers, plus the lost productivity from absenteeism, premature retirement and death. PMID- 15344304 TI - Cost effectiveness of human immunodeficiency virus postexposure prophylaxis for healthcare workers. AB - OBJECTIVE: The United States Public Health Service (USPHS) published recommendations for human immunodeficiency virus (HIV) postexposure prophylaxis (PEP) of healthcare workers in May 1998. The aim of this study was to analyse the cost effectiveness of the USPHS PEP guidelines. DESIGN AND SETTING: This was a modelling study in the setting of the US healthcare system in 1989. The analysis was performed from the societal perspective; however, only HIV healthcare costs were considered and health-related losses of productivity were not included. METHODS: A decision tree incorporating a Markov model was created for 4 PEP strategies: the current USPHS recommendations, triple drug therapy, zidovudine monotherapy or no prophylaxis. A probabilistic sensitivity analysis using a Monte Carlo simulation was performed. Confidence intervals (CIs) around cost effectiveness estimates were estimated by a bootstrapping method. RESULTS: The costs (in 1997 US dollars) per quality-adjusted life-year (QALY) save by each strategy were as follows: monotherapy $US688 (95% CI: $US624 to $US750); USPHS recommendations $US5211 (95% CI: $US5126 to $US5293); and triple drug therapy $US8827 (95% CI: $US8715 to $US8940). The marginal cost per year of life saved was: USPHS recommendations $US81 987 (95% CI: $US80 437 to $US83 689); triple drug therapy $US970 451 (95% CI: $US924 786 to $US 1 014 429). Sensitivity testing showed that estimates of the probability of seroconversion for each category of exposure were most influential, but did not change the order of strategies in the baseline analysis. With the prolonged HIV stage durations and increased costs associated with recent innovations in HIV therapy, the marginal cost effectiveness of the USPHS PEP strategy was decreased to $US62 497/QALY saved. All 3 intervention strategies were cost effective compared with no postexposure prophylaxis. CONCLUSIONS: Current USPHS PEP recommendations are marginally cost effective compared with monotherapy, but the additional efficacy of triple drug therapy for all risk categories is rewarded by only a small reduction in HIV infections at great expense. For the foreseeable future, assuming innovations in therapy that employ expensive drug combinations earlier in the HIV disease course to extend life expectancy and the increasing prevalence of HIV drug resistance, our model supports the use of the USPHS PEP guidelines. PMID- 15344305 TI - Cost effectiveness of cephalosporin monotherapy and aminoglycoside/ureidopenicillin combination therapy. For the treatment of febrile episodes in neutropenic patients. AB - OBJECTIVE: To assess the relative cost effectiveness of cephalosporin monotherapy options and aminoglycoside/ureidopenicillin combination therapy for the treatment of febrile episodes in adult patients with neutropenia. DESIGN AND SETTING: This was a retrospective cost-effectiveness analysis conducted from the institutional perspective. METHODS: The analysis was based on 741 febrile episodes in adult patients with neutropenia enrolled in 5 randomised trials: 3 comparing monotherapy with ceftazidime or cefepime, and 2 comparing cefepime monotherapy versus aminoglycoside/ureidopenicillin combination therapy. Resource utilisation included costs for study antibacterials, treatment of adverse effects and failures, and hospitalisation. The primary end-point was the overall cost of treatment per patient. Cost-effectiveness ratios were also analysed. RESULTS: No significant differences in clinical success rates were detected. Median per patient costs in the monotherapy comparisons were $US7849 for cefepime and $US7788 for ceftazidime [1997 values; not significantly different (NS)]. Corresponding costs for the monotherapy versus combination therapy comparisons were $US9780 for cefepime and $US10 159 for gentamicin/ureidopenicillin (NS). Despite a higher acquisition cost for cefepime, there were no statistically significant differences in cost effectiveness compared with either ceftazidime monotherapy or gentamicin/ureidopenicillin combination therapy. Sensitivity analyses revealed that monotherapy can be cost effective compared with combination therapy in many situations. CONCLUSION: There were no economic differences between the 3 regimens tested. Therefore drug cost should not be a deciding factor when choosing antibacterial therapy for the treatment of febrile episodes in adult patients with neutropenia. PMID- 15344306 TI - Cost effectiveness of acute imipramine therapy versus two imipramine maintenance treatment regimens for panic disorder. AB - OBJECTIVE: To examine the medical costs and effectiveness of acute treatment with imipramine versus acute treatment plus 2 different maintenance therapies for panic disorder. METHODS: A clinical decision model was constructed to estimate 18 month costs and outcomes associated with these treatment scenarios based on the medical literature and clinician judgment. The clinical parameters and outcomes for the model were derived from a series of systematic clinical trials with imipramine utilising uniform dosage procedures and validated response criteria. Costs were calculated based on standardised treatment regimens. The outcome measures were 18-month medical costs, quality-adjusted life years (QALYs) and costs per QALY gained. A sensitivity analysis was performed to explore the impact of treatment withdrawals on outcomes. STUDY PERSPECTIVE: US mental healthcare system. RESULTS: Over 18 months, the total costs (1997 values) and QALYs associated with half-dose maintenance therapy (imipramine 1.1 mg/kg/day) [$US3377; QALYs = 0.991] and full-dose maintenance therapy (imipramine 2.25 mg/kg/ day) [$US3361; QALYs = 0.991] were almost identical; both were cost saving compared with acute imipramine therapy (2.25 mg/kg/day) with no maintenance treatment ($US3691; QALYs = 0.979). Whether patients withdrawing from treatment were considered to have continued to respond to treatment or to have relapsed, the half-dose and full-dose maintenance treatments were still cost saving compared with acute treatment alone. CONCLUSIONS: The results indicate that imipramine maintenance treatment is cost effective compared with acute imipramine treatment for patients with panic disorder. The basic findings and conclusions are not affected after modifying model assumptions for clinical response in patients withdrawing from treatment. PMID- 15344307 TI - The cost effectiveness of antiretroviral regimens for the treatment of HIV/AIDS. AB - OBJECTIVE: To estimate survival, the number of life-years gained and cost effectiveness of antiretroviral therapy (ART) regimens, denoted as ERA-I [zidovudine + (didanosine or zalcitabine)]; ERA-II [stavudine + (didanosine or zalcitabine) or lamivudine + (zidovudine or didanosine or zalcitabine or stavudine)]; and ERA-III [2 nucleoside reverse transcriptase inhibitors + (1 protease inhibitor or 1 non-nucleoside reverse transcriptase inhibitor)]. DESIGN: Modelling of drug cost, cost of opportunistic diseases and survival of HIV positive men and women in the province of British Columbia who were first prescribed any ART between October 1992 and June 1996. A 'reference cohort' was modelled upon individuals in a longitudinal cohort of homosexual men followed since 1982. PERSPECTIVE AND SETTING: Third-party payer perspective in British Columbia, Canada. PATIENTS: All HIV-positive men and women aged > or =18 years with CD4+ counts < or =350 cells/microL who were enrolled in the province-wide drug treatment programme. MAIN OUTCOME MEASURES: Annual costs, survival and cost effectiveness ratios of successive ART regimens. RESULTS: Total costs [1997 Canadian dollars ($Can)] at 12 months under ERA-I, -II and -III were $Can4897, $Can6620 and $Can 11 914, respectively. Survival at 12 months under ERA-I, -II and -III was 89.6%, 91.0% and 97.6%, respectively. The annual incremental cost (estimated by the total incremental cost at 12 months) between ERA-II and ERA-I was $Can1723. The incremental cost-effectiveness ratios between ERA-III and ERA I, and between ERA-III and ERA-II were $Can58 806 and $Can46 971 per life-year gained, respectively. CONCLUSION: We found the cost effectiveness of ERA-III ART regimens well within the range of currently funded therapies for the treatment of other chronic diseases. PMID- 15344308 TI - Cocaine medications, cocaine consumption and societal costs. AB - OBJECTIVE: To estimate the benefits of reduced cocaine consumption in terms of reduced societal costs resulting from the introduction of a medication for cocaine dependence with a small incremental treatment effect. STUDY DESIGN: Cost benefit analysis is applied to study the implications of reduced cocaine consumption. A modelling approach extrapolates the magnitude of treatment effects. METHODS: Epidemiological data on cocaine use and consumption as well as economic methods of cost-benefit analysis are utilised. Estimates of societal costs associated with heavy users of cocaine, who are most likely addicted and in need of immediate treatment, are developed using 1995 data. MAIN OUTCOME MEASURES AND RESULTS: In the first analysis, a postulated 1% reduction in consumption of cocaine among heavy users is examined to approximate a small treatment effect, resulting in a minimal consumption benefit. It is estimated that such a reduction would be valued at $US259 million. The cost-benefit analysis indicated that a cocaine medication with a small treatment effect (10 percentage point increase in abstinence rates) would result in a benefit to cost ratio in the range of 1.58 to 5.79, depending on prescribing behaviour and type of patient. CONCLUSIONS: Such estimates of the benefits of these small treatment effects are conservative, and they may be biased downwards since the willingness to pay for such a cocaine medication could far exceed the benefit to cost estimation used in this paper. Nevertheless, the substantial benefits found in this paper indicate how important investment in cocaine medication is for public health policy; costs may be reduced with efficient prescribing behaviour. Market and governmental barriers to the utilisation of a cocaine medication could reduce the benefits and increase costs. Clinical trials, cost-effectiveness studies, and cost-benefit studies must be conducted to establish the actual pattern of benefits and costs that could be obtained for an efficacious and effective cocaine medication. PMID- 15344309 TI - [Aberrant development of pollen in transgenic tobacco expressing bacterial iaaM gene driven by pollen- and tapetum-specific promoters]. AB - Microsporogenesis offers an ideal model for studying gene expression, cell division and cell to cell communication during development. The role of auxin in pollen development was investigated in transgenic tobacco plants expressing the coding region of the iaaM gene from Pseudomonas syringae, under control of the promoters Lat-52 (pollen-specific) or TA-29 (tapetum-specific). IAA level in anther of transgenic plants increased significantly, and transgenic plants displayed morphological aberrations not solely attributable to pollen development(such as adventitious root formation on stems, epinastic leaf growth, delayed flowering). These results suggest that expression of Lat-52 and TA-29 are not strictly limited to anther. Anther shape was changed and the number of pollen grains per anther was reduced, but grains could be stained with aceto-carmine. Almost all flowering plants were fertile, although the number of flowers per inflorescence was reduced compared with the wild-type ones. These results suggest that auxin plays an important role in pollen development, and over-expressing auxin synthesis gene could result in aberrant development of pollen. PMID- 15344310 TI - [Location studies of Smad4 protein in the rat testis during postnatal development]. AB - In order to shed light on the mechanisms of TGF-beta action in the testis,we examined the expression and function of Smad4 protein, the common-mediator Smads, which is one of intracellular signaling molecules of TGF-beta superfamily members, in rat testis during postnatal development. Whole testes were collected from SD rats aged 3 days, 7 days, 14 days and 28 days, and adult. In this study, we examined, by means of western blots, the protein expression of Smad4 during rat testicular development and its cellular localization by immunohistochemical ABC method with glucose oxidase-DAB-nickel enhancement technique. The results showed that the protein of Smad4 was present in rats from 3 days of age to adulthood, and the immunoreactivity for Smad4 was exclusively localized to the cytoplasm of Leydig cells with negative nuclei in the interstitial tissue at any time point. No expression was detected in germ cells. Therefore, our data provide evidence for the molecular mechanism of TGF-beta action in rat testes during postnatal development and spermatogenesis of rats. PMID- 15344311 TI - Immunocytochemical localization of estrogen receptor beta in the rat brain. AB - The localization of the new estrogen receptor, ER-beta, in the rat brain was studied by immunocytochemical technique, and the results revealed that ER-beta immunoactive material was predominantly localized in the neuronal nucleus, but it was also detectable in the cytoplasm and neuronal processes. High levels of ER beta immunopositive signals were detected in the cerebral cortex, vertical limb of the diagonal band, Purkinje cells, locus ceruleus, and motor trigeminal nucleus. Moderate levels were found in the medial septum, lateral amygdaloid nucleus, substantia nigra, and central gray. Weak signals were localized in some subregions of the hypothalamus and amygdaloid complex. Some differences of the expression of ER-beta immunoreactivity between male and female rats were also noticed. The above results provide the first detailed evidence that ER-beta protein is widely distributed in the rat brain, and ER-beta may be involved in some important brain function such as learning and memory. PMID- 15344312 TI - [Characterization of expression of the centrosomal protein, Cenexin, in rat spermatogenesis]. AB - The centrosomal protein, Cenexin, is a molecular marker of mature centriole. To elucidate the variability and function of mature centriole in spermatogenesis, the high titer polyclonal antibody against rat cenexin was obtained by immunizing mice with recombinant cenexin which was made up in this study. The expression of cenexin in rat spermatogenesis was carried out by semi-quantitative RT-PCR, immunofluorescence and Western Blot. The results demonstrated that the level of Cenexin mRNA was higher in spermatogonia and spermatocytes, then decreased in following stages, while Cenexin protein was located on one centriole from spermatogonia to spermatids, showing mature centriole existed in these stages. Cenexin protein was localized to the basal body of the flagellum in elongated spermatids and the stained signal disappeared in the most of epidydimal sperms. These results suggested that the expression pattern of cenexin in rat spermatogenesis might be related to the initiation of the flagella formation. PMID- 15344313 TI - [Changes of thioredoxin mRNA level in neurons insulted by sodium azide]. AB - Oxidative stress has been considered to be associated with many neurodegenerative diseases, while mitochondrial damage is one of the important reasons that aggravate oxidative stress. Sodium azide (NaN3) is a specific inhibitor of mitochondrial cytochrome c oxidase (COX), which can be used to mimic neuronal damage induced by mitochondrial deficiency. In this experiment, the neurotoxic effects of NaN3 on cultured primary neurons were detected by means of cell viability measurement (MTT assay) and morphological observation, and an in vitro model of neuronal injury induced by NaN3 were established. Further the changes of thioredoxin mRNA level in cultured neurons insulted by NaN3 were analyzed by semiquantitative RT-PCR in order to explore the role of Trx, an important redox regulatory protein, in the process of neuronal injury. It was found that cultured neurons could be damaged by NaN3 in a dose- and time-dependent manner and the expression of Trx decreased during certain dose (0-32mmol/L) and time (0-4hr) of NaN3 treatment. The above data suggest that decreased the expression of Trx may be associated with oxidative stress induced by neuronal respiratory chain injury, which could impair neuronal protective mechanism. PMID- 15344314 TI - [Isolation of 5' flanking region of the fruit-specific gene wml1 from watermelon by Uneven PCR]. AB - The 5' flanking region of ADP-Glucose Pyrophosphorylase large subunit gene wml1, which covers 1864 bp, was isolated from watermelon genomic DNA by Uneven PCR. Sequence analysis indicated that there were putative TATA box and CAAT box at position 1237bp and 1284bp of the cloned fragment, respectively. Two transcriptional fusions between 5' regulational sequence of wml1 and beta glucuronidase gene were constructed. Constructs containing 180bp-1752bp fragment could drive GUS gene transient express specifically in watermelon fruit, while 958bp-1752bp had no promoter activity. Putative fruit-specific cis-acting elements may locate between 180bp and 958bp. PMID- 15344315 TI - [Cloning and analysis of highly repetitive sequence fragments from takin (Budorcas taxicolor)]. AB - Takin (Budorcas taxicolor) is a large animal living in China and other adjacent countries, which belongs to Bovidae of Artiodactyla. The anatomy, morphology and behavior of takin are between species of the subfamily Bovinae and Caprinae. It is now in a separated genus or in the same genus with muskox (Ovibos moschatus). To gain insight into the evolution of takin, we purified and cloned its highly repetitive BamHI fragments from the genomic DNA. The sequences of three fragments were highly homologous, indicative of units of a large repetitive DNA arrays. Southern hybridization using these fragments as probes showed identical patterns among individuals in both the same and different subspecies, implying conserved sequences and distributions of the BamHI clusters in takin genome. Sequence comparison with 1.714 and 1.715 satellite DNA of other species in Bovidae supports that takin has a closer relationship with the subfamily Caprinae than with Bovinae. It also suggests that these BamHI fragments may represent repeat units of the centromeric satellite DNA of takin. PMID- 15344316 TI - [Transferring of multiple insect-resisting genes into two-line genetic male sterile rice Pei'ai 64S by particle bombardment]. AB - Transformation of two-line genetic male sterile indica rice variety Pei'ai 64S was conducted by particle bombardment. Three insect-resisting genes ligased into plasmid vector pKC-3 was introduced into calli derived from mature embryo and a total of 33 transgenic plants had been obtained. The PCR and Southern blot analysis of each different gene in R0 plants and also PCR analysis of each different gene in R1 plants showed the integration of three insect-resisting genes into the genome of transgenic plants which could stably pass down. PMID- 15344317 TI - [The ultracytochemical localization of ATPase activity in pollen tube and stigma of Fagopyrum esculentum after compatible and incompatible pollination]. AB - Ultracytochemical localization of adenosine triphosphatase (ATPase) activity in stigmas, pollens and pollen tubes of Fagopyrum esculentum was performed with the cytochemical method of lead phosphate precipitation. The results were as follows: (1) Lower activities of ATPase appeared in stigma cells at 0.5 hour after compatible and incompatible pollination. Stigma surface and pollen grains attached on stigma showed higher ATPase activities after compatible pollination and lower or no activities after incompatible pollination at 0.5 hour. ATPase localized on endoplasmic reticula and sperm cell in pollen grain. (2) Lower ATPase activities appeared both in stigma cells and pollen tubes in style at 1.5 hours after incompatible pollination. Pollen tube stopped growing and the degeneration of its cytoplasma began; on the contrary, at 1.5 hours after compatible pollination, higher ATPase activities were detected both in stigma cells and pollen tube in style. ATPase localized mainly on plasmolemma, in cytoplasmic matrix of stigma cells and in mitochondria, dictyosome, plastid envelop, and on the wall of pollen tube as well. The present study indicated that the stop of pollen tube growth after self-incompatibility of Fagopyrum esculentum resulted not only from the nutrient starvation of pollen tube, but also from its metabolic disorder in incompatible style. PMID- 15344318 TI - [Effects of simulated microgravity on preimplantation embryonic development of Kunming mouse in vitro]. AB - In order to investigate the development of Kunming mouse preimplantation embryos cultured in vitro under simulated microgravtiy, one-cell and 4-cell embryos of Kunming mouse (Fertilization In Situ) were cultured in CZB or KSOM media under simulated microgravity or normal gravity environment respectively. The results showed that under normal gravity, the percentage of passing 2-cell development block embryos was not different in CZB with in KSOM, but the percentage of blastocysts was lower in CZB than that in KSOM significantly. The percentages of passing 2-cell development block embryos and blastocysts in CZB or KSOM were lower under simulated microgravity than those under normal gravity. It is suggested that the frequency of early embryonic lethality is possibly increased by simulated microgravity. PMID- 15344319 TI - [In silico cloning of evolutionarily conserved mouse F-LANa]. AB - Differentially expressed genes between normal liver and hepatocellular carcinomas were investigated using differential display. In previous study, human F-LANa was identified as a differentially expressed gene, up-regulated in hepatocellular carcinoma. In this study, we developed an in silico cloning approach to rapidly and accurately characterize the mouse ortholog of the human F-LANa. Mouse F-LANa encodes a 239 aa protein exhibiting 97.9% similarity to the human ortholog gene. Homology analysis was carried out in various species and showed that F-LANa was evolutionarily conserved from yeast to human. Based on the alignment results, phylogenetic tree was established here. PMID- 15344320 TI - [Interspecies nuclear transfer using nonquiescent somatic cell]. AB - It is the point at issue in intraspecies nuclear transfer whether quiescence is necessary for development of nuclear transfer reconstructed embryos. In the interspecies nuclear transfer, some reports have proved that quiescent cell is able to support preimplantation development of the interspecies reconstructed embryos. Are non-quiescent cells able to support preimplantation development of the interspecies reconstructed embryos? We used non-quiescent somatic cells from C57BL/6 mice and giant pandas as donors to transfer into enucleated rabbit oocytes. After electrofusion (the electrofusion rates were 62.2% and 71.6%, respectively) and electrical activation, 5.1% of those mouse-rabbit reconstructed embryos developed to blastocyst in vitro, and 4.2% of panda-rabbit reconstructed embryos developed to blastocyst after transferring into ligated rabbit oviduct. These results indicate that non-quiescent cell from C57BL/6 mouse and giant panda could be dedifferentiated in enucleated rabbit oocytes and support early embryo development. PMID- 15344321 TI - [Helical structure revealed upon the beta heterochromatin of polytene chromosome of Drosophila virilis]. AB - By using conventional electron microscopy, beta heterochromatin of polytene chromosome of Drosophila virilis was composed of 80-100nm chromonemas. Helical structures consisting of the 30nm chromatin fibres were identified in the chromonema. Based on these observations, the radial loop model and the multiple coiling model were discussed. PMID- 15344322 TI - [Ultrastructural localization of transcription sites of rRNA genes in the nucleolus of wheat]. AB - There were no definite conclusions about transcription sites of rRNA genes in nucleoli of eukaryotes for a long time. We used the wheat as specimen and observed ultrastructure of chromatin in FC of nucleoli by conventional transmission electron microscope. And we employed anti-DNA antibodies to analyze the distribution of DNA in nucleoli, and found that DNA located in FC, DFC and the transitional region between them. Using anti-UBF antibodies to study localization of RNA polymerase I transcription factor UBF in nucleoli, we observed that UBF located in DFC and the transitional region between DFC and FC in wheat, but not in FC. Furthermore, with anti-RNA/DNA hybrid antibodies, we directly and selectively labeled the transcription sites of rRNA genes of nucleoli and revealed that it located in DFC and the transitional region between DFC and FC in wheat. PMID- 15344323 TI - [Expression and characterization of carboxylesterase A2 in E. coli]. AB - The most commonly observed change that has been linked to resistance development is the increase in activity of carboxylesterases. The putative mechanism involves an overproduction of this enzyme for the sequestration and the hydroxylation of various organophosphate and carbamate insecticides. Carboxylesterases A2 cDNA was amplified from Culex quinquefasciatus by RT-PCR and sequenced consequently. Target gene was inserted into pET-28a to create prokaryotic expression plasmid pET-EstA2. When pET-EstA2 was transformed into E. coli BL21, the recombinant was induced by IPTG. A pure recombinant protein was obtained by affinity purification. Compared with carboxylesterase A2 purified from Culex quinquefasciatus, carboxylesterase A2, purified from the product of the transgenic of E. coli, has the same Km, but the Vm was higher than that of it, which shows that carboxylesterase A2, purified from the product of E. coli by affinity, is purer than that from Culex quinquefasciatus. The study on the expression and characterization of carboxylesterase A2 in E. coli is more useful for its future application. PMID- 15344324 TI - [Study on the mechanism of inhibition of tumor cell motility by ascorbic acid derivatives]. AB - Our previous study shows that tumor invasion is inhibited by 2-O-phosphorylated ascorbate-6-O-palmitylester (Asc2P6Plm). In the present study, the mechanism underlying the inhibitory effect of Asc2P6Plm on invasion of human fibrosarcoma cells HT-1080 was attempted to be analysed. Migratory ability of the tumor cells was shown to be inhibited in a dose-dependent manner by treatment with Asc2P6Plm at 50-300 micromol/L for 1 hr. Hydroxyl radicals in homogenates of Asc2P6Plm treated HT-1080 cells were markedly diminished relative to those of non-treated cells as evaluated by electron spin resonance method using the spin-trapping agent DMPO. F-actin was localized in the vicinity of the cell membrane abundantly in nontreated cells, but was diminished in a time-dependent manner in Asc2P6Plm treated cells as shown with the F-actin-directed agent NBD-phallacidin. The cell adhesion-controlling molecule RhoA increased time-dependently in the cell nucleus of Asc2P6Plm-treated cells as shown by Western blots. Thus the inhibition of tumor invasion by Asc2P6Plm was shown to be attributed to decreases in both the cell migratory ability and the F-actin localization near the cell membrane, which may result from an increase in RhoA in the cell nucleus and reduction of intracellular ROS that is achieved by enrichment of intracellular Asc derived from Asc2P6Plm. PMID- 15344325 TI - [Expression of monoclonal nonspecific suppressor factor beta in E. coli, preparation of its antibodies and its tissue orientation in the mouse endometrium]. AB - In order to explore the role of MNSFbeta in the process of implantaion, MNSFbeta and its antibodies are required. The expression plasmid pBV220/MNSFbeta-hCGbeta was constructed, and then transferred into E. coli to express the fusion protein MNSFbeta-hCGbeta. The anti-hCGbeta antibody was used to identify the fusion protein. The result demonstrated that MNSFbeta-hCGbeta was expressed correctly and its molecular weight was consistent with the anticipated one. Finally the expression product MNSFbeta-hCGbeta was preliminarily purified and used to immunize Balb/C mouse to generate the antibodies. In the meantime, the expression plasmid pGEX-4T-2/MNSFbeta was also constructed and transferred into E. coli to express the fusion protein GST-MNSFbeta. GST-MNSFbeta was purified and used to stimulate the immunized mouse before the preparation of hybridomas cells. The prepared polyclonal and monoclonal antibodies against MNSFbeta were checked and measured by fusion protein GST-MNSFbeta. The prepared polyclonal antibody was then used to perform the immunohistochemistry analysis. The result suggested that the level of MNSFbeta in interimplantation sites was significantly higher as compared with implantation sites in the mouse uterine on Day 4.5 of pregnancy. PMID- 15344326 TI - [Double action of dopamine on PC12 cells]. AB - PC12 cell line was used to examine the cytotoxicity of dopamine (DA). The higher dose of DA (400micromol/L) greatly reduced cell survival rate and induced apoptosis,meanwhile the lower dose (100micromol/L) had less cytotoxicity. Further studies showed that AMP (100micromol/L) were not cytotoxic at 24h, but decreased cell survival rae to 78% at 72h compared to control, and also exacerbate the cytotoxicity of DA (100micromol/L) at 72h. By determine the action of GSH level and SOD activity, we found that appropriate intracellular DA may be advantageous to protect cells from the toxicity of higher dose DA. The intracellular calcium concentration showed that DA's toxicity increased intracellular concentration of calcium. The results suggest that it was the extracellular DA that made cytotoxicity by decreasing cell's possibility of antioxide and increasing intracellular free calcium. PMID- 15344327 TI - [Sheep transgenic embryos produced by intracytoplasmic sperm injection]. AB - In this study, the possibility of sheep transgenesis by intracytoplasmic sperm injection (ICSI) was assessed. In experiment 1, activation of ovine oocytes matured in vitro in preparation for ICSI has been investigated with 3.42 mmol/L Ca2+ treatment, ionomycin alone and ionomycin followed by 6-dimethylaminopurine (DMAP) after 3-h delay (group 1, 2 and 3, respectively). After activation, the oocytes were then cultured in SOFaaBSA medium. Cleavage rates were significantly (P<0.05) different among three groups (18.4%, 91.8% and 71.7%, respectively). In additional culture, no parthenotes in group 1, whereas 11% and 17.4% in group 2 and 3 developed to the blastocyst stage. Therefore we used the third activation method in the following ICSI tests. In experiment 2, development of ovine oocytes after ICSI was investigated. Thawed semen from two rams was separated by Percoll centrifugation and was used for ICSI or in vitro fertilization (IVF) trails. A total of 71.8% of oocytes reached the 2-cell stage following living sperm injection, which was significantly (p>0.05) different from those following IVF (41.4%) and sham-ICSI (30.2%). After seven days' culture, no sham-injected oocytes developed into the blastocyst stage, although 7% in ICSI and 16.1% in IVF oocytes developed into the blastocyst stage, but there was no significant difference in ICSI and IVF groups (p>0.05). In the further study, the possibility of sheep transgenesis by ICSI was assessed. After coinjection of ovine oocytes matured in vitro with dead sperm cold to -20 degrees C and exogenous DNA encoding green fluorescent protein (GFP), seventy-three percent of coinjected oocytes developed to 2-cell stage (33/45) and two of them were transgene-expressing embryos. Among ten embryos at the 16-cell stage, all embryonic cells in one transgenic embryo still expressed GFP. Four coinjected blastocysts were thawed and transferred to the uterine of the two progesterone-synchronized recipient ewe. No pregnancies were detected on the 60th day. These results suggested sheep transgenic embryos could be produced by ICSI and further studies should be performed. PMID- 15344328 TI - Ultrastructural localization of ATPase in mature tracheary elements of rice (Oryza sativa L.) stamen. AB - The development of mature tracheary elements (TEs) of rice (Oryza sativa L. cv. Nongkeng 58s-SD) stamen from big-vacuole to trinucleate pollen stage was studied with lead phosphate--deposition technique. At big-vacuole pollen stage, mature TEs, both in filament and connective, were short of ATPase activity. However, ATPase activity appeared at binucleate pollen stage; in filament, one or two vascular bundle cell (s) next to TE degenerated earlier than the rest; and the plasma membrane of degenerating cell (DC) always exhibited high ATPase activity on condition that the adjoining TE was of fully stretched second wall; when the second wall only turned up far off DC, the plasma membrane of DC still showed high ATPase activity except on borderline between DC and TE; in connective at the same stage, TEs also exhibited high ATPase activity, but the vascular bundle cell(s) adjoining TEs degenerated much later than their counterparts in filament. At trinucleate pollen stage, materials, either with or without ATPase activity, were observed both in filament and connective TEs. The above results indicated that (some of the) materials in mature TEs come from filament DC. PMID- 15344329 TI - [Transformation of Bt-CpTi fusion protein gene to cabbage (Brassica oleracea var. capitata) mediated by Agrobacterium tumefaciens and particle bombardment]. AB - Bt-CpTI fusion protein gene was transferred to the explants of hypocotyl, cotyledon with petiole and shoot apex of cabbage (Brassica oleracea L. var. capitata) variety "Zhonggan No 8" via Agrobacterium tumefaciens and particle bombardment, and 13 kanamycin-resistant plants were obtained. PCR and Southern blotting hybridization verified that all these plants of the kanr plants of type I regenerated from hypocotyl and cotyledon with petiole mediated by A. tumefaciens were transgenic plants, 2 karr plants of type II stemed from shoot apex mediated by A. tumefaciens were "false-positive"plants and one of 2 kanr plants of type III regenerated from shoot apex via particle bombardment was non transformed plants. It was showed that part of transgenic plants had high activity of trypsin inhibitor and strong resistance to resist common cabbage worm through the analysis of CpTI relative capacity and insect-resistant test. PMID- 15344330 TI - [Study on the editing sites of the coxII gene transcripts of rice mitochondria]. AB - The term RNA editing is generally used to describe those molecular processes in which the information content is altered in an RNA molecule. This process is not limited to mRNA since alterations of non-informational RNA have also been found. RNA editing exists extensively in the higher plant mitochondria, and is the necessary step for forming functional proteins. In this paper, the research materials are the gametopthyte male sterility line (A), maintainer line (B) and F1 hybrid (F1) of HL-type CMS. 15 editing sites are found in the transcripts of coxII by comparing cDNAs and DNAs sequences. A,B and F1 have same Editing sites. When editing occurs at the first or second position of codons, the encoded amino acid is likely to be altered. As a result, the conservation of the predicted protein is improved as compared with other organisms. PMID- 15344331 TI - [Investigation of nuclear Ca2+ regulation in the isolated cardiac nuclei]. AB - To investigate the regulation of Ca2+ in the isolated cardiac nuclei from rats which may illuminated the mechanism of nuclear calcium transport system. Elocity and isopyknic gradient centrifugation were employed to fractionate rat cardiac nuclei. Then fluo-4 confocal microscopy techniques was used to verify the changes of nuclear Ca2+. There are calcium-dependent Ca2+ uptake in the cardiac nuclear obtained from normal rats. The accumulation Ca2+ of cardiac nuclei in vitro from the incubating medium were not consistent with free [Ca2+] in incubating medium. The nuclear envelope was initially loaded with Ca2+ (1 mmol/L ATP and approximately 100 nmol/L Ca2+), Adequate Ca2+ loading was next confirmed by imaging the nuclear envelope and nucleoplasm. Exposure of Ca2+ -loaded nuclei to IP3, ryanodine or ryanodine + thapsigargin, respectively, resulted in a rapid and transient elevation of nucleoplasmic Ca2+ free concentration, this effects were abolished by pretreatment of cardiac nuclei with Ca2+ -ATPase inhibitor thapsigargin. Thapsigargin and IP3 receptor antagonist heparin induced nucleoplasmic Ca2+ free concentration decrease. Fluorescence experiments indicated that both ryanodine receptors and Ca2+ -ATPase were distributed in the outer layer of nuclear envelope, and inositol 1,4,5-trisphosphate receptors mainly dispersively localized at inner layer of nuclear envelope. The present study demonstrates that nuclear calcium were regulated by free Ca2+, IP3 and ryanodine. The results suggested calcium transport system might be present in the myocardial nuclei, the myocardial nuclei might served as one of calcium pools in myocardial cell. PMID- 15344332 TI - [Cloning and identification of a mouse zinc finger protein gene ZF-12-related pseudogene]. AB - The mouse zinc finger protein ZF-12 gene is homologous to human gene and encodes a protein of 368 amino acids, which contains four tandem C2H2-type zinc finger motifs in the N-terminal and one SCAN domain in the C-terminal. Some recent studies suggest that ZNF191 might be a hepatocarcinogenesis-associated gene. We screened a mouse lambda genomic library with a human ZNF191 cDNA probe and isolated a ZF-12-like gene, named ZF12p (GenBank AY040222). This intronless gene closely resembles ZF-12 but displays several mutations, suggesting that ZF12p represents a ZF-12-related pseudogene. RT-PCR analysis on total RNA from mouse tissue and bioinformatis analysis on promoter region of ZF12p gene, suggest the transcripts of ZF12p may be not synthesized. BLAST on the data of the human genome in the GenBank with ZNF191 cDNA and Southern blotting show there is no any psedogene related to ZNF191 gene in the human genome. The high similarity of ZF12p to ZF-12 might be of considerable importance for mutation and evolution analysis of ZF-12. PMID- 15344333 TI - [Isolation and culture of human pluripotent embryonic germ cells]. AB - To establish human pluripotent embryonic germ (EG) cell lines, human primordial germ cells (PGCs) of embryos aborted in 5-9 week were cultured on inactive mouse STO fibroblast feeder. The medium contained human leukemia inhibitory factor (hLIF), human basic fibroblast growth factor (hbFGF) and forskolin. The EG cells could be passaged continuously until 12 generations. Most cells were positive in alkaline phosphatase staining and expressed cell surface antigen SSEA-3 and pluripotent marker Oct-4. These EG cell populations that retained normal karyotype could form embryoid body in culture and differentiate further into neuron-like cells, mucous epithelial cells, epithelial cells and other types of the cells spontaneously. These results indicated the cell clones derived from human PGCs resemble pluripotent EG cells from mouse PGCs in appearance or nature. PMID- 15344334 TI - [Studies on calcium sensitive period during floral differentiation of cucumber cotyledonary node cultures in vitro]. AB - Cotyledonary nodes of cucumber cultured on calcium-free medium for 0, 1, 2, 3, 4, 5, 6d respectively, were transferred to medium with 6.0 mmol/L CaCl2 for 24h, then returned to calcium-free medium. Cotyledonary nodes cultured on calcium-free or 6.0 mmol/L CaCl2 medium for all time, were taken as controls. Results showed that cotyledonary nodes were transferred to 6.0 mmol/L CaCl2 medium for 24h during 0-3d after the beginning of culture, percentage of floral bud formation at cotyledonary nodes was increased significantly. Transferring cotyledonary nodes on the 3d day after the beginning of culture was achieved best effect, percentage of floral bud formation was up to 34.3%. We deduced that the calcium sensitive period during floral differentiation of cucumber cotyleddonary node cultured in vitro may be 0-4d after the beginning of culture. PMID- 15344335 TI - [Effects of salinity stress on cytosolic calciumin Stevia rebaudiana Bertoni cells]. AB - Fluo-3/AM, a calcium indicator, was introduced by low temprature loading method into callus protoplasts of Stevia rebaudiana Bertoni. The microscope observation results showed that NaCl in different concentrations (30-200mmol/L) can elevate the intracellular calcium concentration in protoplasts. The elevation was related to the amount of salinity. The effect of salinity stress was inhibited by LiCl pretreatment but restored by inositol pretreatment. This suggested that salinity stress promoted the cytoplasmic calcium activity, perhaps by activating the phosphoinoditide system. As a result of salinity stress signals, elevated calcium activity may trigger corresponding metabolic changes, such as synthesis of enzyme via activating other members of calcium signal system, fit the cells for the change of the environment. PMID- 15344336 TI - [Bcl-2 expression in enteric neurons of Hirschsprung's disease and its significance]. AB - The bcl-2 protein, which widely expressed in the developing central and peripheral nervous systems, has the functional role of blocking apoptosis. The purpose of this study was to map bcl-2 expression in the human enteric nervous system and investigate the value of bcl-2 immunohistochemical method in the diagnosis of Hirschsprung's disease (HD), as this has not previously been done. Rectal specimens were obtained from definitive operation of 20 patients with HD. Specimens were analyzed with immunohistochemical methods, using antibodies against bcl-2. The bcl-2 protein was expressed in myenteric and submucous neurons in normal adult and HD expand segment, but no bcl-2 immunoreactive enteric neurons was revealed in the narrow segment. And nerve fibers of the enteric plexuses that were bcl-2 immunoreactive were few in all examined specimens. From the conclusion, expression of bcl-2 is displayed in enteric neurons and immunohistochemical analysis of bcl-2 may also be valuable for identification of the enteric neurons in HD. PMID- 15344337 TI - [Changes in number and distribution of retinal ganglion cells after optic nerve crush in zebrafish]. AB - Changes in number and distribution of retinal ganglion cells were studied after optic nerve crush in zebrafish (Brachydanio rerio) with retinal wholemount. There were approximately 40,000 to 56,000 cells in the retinal ganglion cell layer. The density of ganglion cells was divided into six classes and the area of highest cell density (central area) was located at the temporal area to the optic disc in normal fish. At the early regeneration stages after optic nerve crush, the percentage of lost cells increased gradually. Cell density had fallen first in the central area. At the late regeneration stages, there was an approximately 20% loss of ganglion cells during optic nerve regeneration. The results suggest that the loss of cells may undergo apoptosis rather than necrosis. A wave of cell loss started in the central area and spread progressively further into periphery. The reason caused these changes may be due to temporal interruption of optic nerve function, recovery from crush and the ability to quickly regenerate in optic nerve of the fish. PMID- 15344338 TI - Advance directives. Good, bad or indifferent. AB - The author writes about various alternatives once decisional capacity is lost. So called advance directives come in two forms: the living will and an appointed proxy for health care. The US--were these have been legally binding for over 20 years in all states--is a useful laboratory for studying the effect. Advantages and disadvantages are discussed. In some civilizations decisions about all such matters are made in different ways--these are not superior or inferior but simply different. In the US with its shameful social conditions for a large part of the population more important problems need to be addressed. In civilizations which have at least basic medical care for all, discussing such documents becomes far more important. The EU is likely to make some form of advance directive have legal standing. The last 20 or more years that they have been utilized in the US may provide experience important for the time when the EU also makes such documents legally binding. PMID- 15344339 TI - Advance directives in the primary care setting. AB - Advance directives have been available for more than 20 years, yet only 2% of patients report having had a discussion about them with their physician. Physicians and patients appear to be reluctant to bring up the subject despite evidence that patients not only want help with advance directives, but report more satisfaction with their health care when the topic is addressed. The primary care setting is particularly well-suited to the establishment of advance directives. A clearer understanding of the benefits of advance directives to physicians and their patients can hopefully increase the use of this important health care resource. PMID- 15344340 TI - Current legal status of advance directives in the United States. AB - Advance directive legislation has been in place throughout much of the United States for nearly 3 decades. The right to give an informed consent to or refusal of medical treatment has been recognized by state and federal courts, and that right has been determined to survive the loss of decisional capacity and may be exercised through the execution of instructional or proxy directives. Despite these developments, the percentage of the adult population with a formal advance directive of any type has never exceeded 15%. Moreover, a remarkable number of these directives are ambiguous and/or their existence is unknown to the physicians who are expected to rely upon them. Even unambiguous directives may not be followed at the critical stage in the trajectory of a patient's illness, and the accountability for such disregard by health care institutions or professionals is negligible to nonexistent. Nevertheless, there is real potential for advance directives, as a key element to sound advance care planning, to fulfill their initial promise as instruments of the prospective autonomy of patients. In order for that potential to be realized, primary care physicians must embrace advance care planning as a part of their professional responsibility to patients. PMID- 15344341 TI - Communication through advance directives. AB - There are two kinds of justifications for advance directives. The first is that they extend patient autonomy into the future, allowing individuals to control or at least influence their care after they have lost the capacity to make decisions. The second is that they help physicians and family members make emotionally stressful decisions in difficult circumstances. Whether directives are written or unwritten is of secondary importance. The primary concern is that they are based on an adequate understanding of medical options. Two cases illustrate the relative importance of these features of advance directives. PMID- 15344342 TI - Advance directives in oncology: are they useful? AB - This paper discusses advance directives from the point of view of advanced directives: living wills, enduring power of attorney and combined instruments. They are discussed from the point of view of patients, the health-care teams and the public. It stresses the need for adequate information before such an instrument can be binding and suggests that enunciating goals is the most important task. PMID- 15344343 TI - Advance directives in an academic setting: what should residents be taught? AB - In this paper I wish to introduce a short description of the US medical educational system and show how various types of Advance Directives can be used in the educational process. The danger is that these documents will become one more thing to be gotten out of the way so that no real discussion takes place. In summation, many academic teaching hospitals, and their faculty are asked to teach residents and students about advance directives, but it is often done in a formulaic way, because the nature of the doctors, the patients, and the medical care system impede true inquiry into the fundamental question which makes an advance directive valuable and legitimate: how can we help this person either live or die in a way that is both medically feasible and closest to their desires? PMID- 15344344 TI - Written advance directives: Theuth's blessing... or... curse? AB - Advance directives are discussed from a philosophical standpoint and with reference to their relationship to informed consent. The necessity of an advance directive being truly informed is stressed. The author suggests that the validity of these instruments is made quite difficult when the particular context in which they find themselves is one which neither patient nor physician anticipated. PMID- 15344345 TI - [Advance directives in Europe]. AB - The types and theories of advance directives in different European countries are reviewed and described. The author carefully compares the legal standing of advance directives in these countries. Since there is no doubt that advance directives in some form will become legal instruments in the EU countries, it is useful to examine how these have worked out so far in some European countries. PMID- 15344346 TI - Panel: law punishing animal rights extremists too weak. New terror tactics circumvent federal statute. PMID- 15344347 TI - O.I.E. takes action at annual meeting. Agency acts on measures involving BSE, zoonoses, animal welfare, and the disease notification system. PMID- 15344348 TI - CDC: rabies transmitted through organ donation. Veterinarians urged to remain vigilant. PMID- 15344349 TI - Special pesticide formulations. Veterinarians must follow EPA rules. PMID- 15344350 TI - Managing stress and avoiding burnout. A self-care primer for overly compassionate and overworked veterinarians. PMID- 15344351 TI - Questions definition of terrorism. PMID- 15344352 TI - Another view of animal friendly curriculums. PMID- 15344353 TI - Warning of an adverse effect of etodolac. PMID- 15344354 TI - Behavior wellness concepts for general veterinary practice. PMID- 15344355 TI - What is your diagnosis? Osteomyelitis of a vertebral body. PMID- 15344356 TI - Distribution of AVMA members by state and gender (1999-2003). AB - From 1999 to 2003, the percentage of actively employed females in the AVMA increased 25.4%, whereas the percentage of males decreased by 0.4%. The number of females increased in all 9 census regions. PMID- 15344357 TI - Methods for analysis of cell-mediated immunity in domestic animal species. PMID- 15344358 TI - Extralabel use of medicated feeds for minor species. PMID- 15344359 TI - Adverse drug event reports at the United States Food And Drug Administration Center for Veterinary Medicine. PMID- 15344360 TI - Evaluation of the effects of premedication on gastroduodenoscopy in cats. AB - OBJECTIVE: To evaluate the effects of hydromorphone, hydromorphone and glycopyrrolate, medetomidine, and butorphanol premedication on the difficulty and time required to pass an endoscope into the stomach and duodenum of cats anesthetized with ketamine and isoflurane. DESIGN: Randomized complete block crossover study. ANIMALS: 8 purpose-bred adult female cats. PROCEDURES: Each cat was premedicated and anesthetized 4 times with an interval of at least 7 days between procedures. Cats were premedicated with hydromorphone, hydromorphone and glycopyrrolate, medetomidine, or butorphanol administered IM. Twenty minutes after premedication, sedation was assessed by use of a subjective ordinal scale. Cats received ketamine administered IM, and 10 minutes later a cuffed orotracheal tube was placed and anesthesia maintained with isoflurane. Cats breathed spontaneously throughout the procedure. When end-tidal isoflurane concentration was stable at 1.4% for 15 minutes, endoscopy was begun. The times required to pass the endoscope through the cardiac and pyloric sphincters were recorded, and the difficulty of endoscope passage was scored by use of a subjective ordinal scale. RESULTS: No significant differences in difficulty or time required to pass the endoscope through the cardiac and pyloric sphincters were found among premedicant groups. Premedication with medetomidine resulted in the greatest degree of sedation and longest time to return to sternal recumbency. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that hydromorphone, hydromorphone and glycopyrrolate, medetomidine, and butorphanol at the doses tested can be used satisfactorily to premedicate cats prior to general anesthesia for gastroduodenoscopy. PMID- 15344361 TI - Naturally occurring tularemia in a dog. AB - A 4-year-old spayed female Irish Setter was examined because of acute onset of lethargy, anorexia, and weakness. The dog had eaten an adult rabbit 36 hours earlier. Tularemia was suspected because of the rabbit exposure; however, other common diseases characterized by fever, malaise, and lymphadenopathy of acute onset were also considered (ie, ehrlichiosis and Rocky Mountain spotted fever). The dog was treated with doxycycline (5 mg/kg [2.3 mg/lb], PO, q 24 h) for 14 days as well as supportive treatment with a balanced electrolyte solution (lactated Ringer's solution [200 mL, SC]). The diagnosis was first established by results of bacteriologic cultures of fine-needle aspirates obtained from lymph nodes and confirmed by results of ELISA and a polymerase chain reaction assay Successful and timely antemortem diagnosis of tularemia in dogs can be accomplished through lymph node aspiration and bacteriologic culture. PMID- 15344362 TI - Evaluation of outcome of otitis media after lavage of the tympanic bulla and long term antimicrobial drug treatment in dogs: 44 cases (1998-2002). AB - OBJECTIVE: To evaluate the outcome of otitis media in dogs after video-otoscopic lavage of the tympanic bulla and long-term antimicrobial drug treatment. DESIGN: Retrospective study. ANIMALS: 44 dogs with otitis media treated in an academic referral practice. PROCEDURE: Medical records were reviewed for signalment, duration of ear canal disease, previous medical treatments, dermatologic diagnosis, results of cytologic examination and microbial culture of ear canal exudate, findings during video-otoscopy, medical treatment, days to resolution, and maintenance treatments prescribed. Four independent variables (age, duration of ear canal disease prior to referral, use of corticosteroids in treatment regimens, and infection with Pseudomonas aeruginosa) were evaluated statistically for potential influence on time to resolution. RESULTS: Mean +/- SD (range) duration of ear canal disease prior to referral was 24.9 +/- 21.6 (3 to 84) months. Otitis media in 36 dogs resolved after lavage of the tympanic bulla and medical management; mean +/- SD (range) time to resolution was 117 +/- 86.7 (30 to 360) days. Time to resolution was not significantly influenced by any variable evaluated. Three dogs were lost to follow-up, and 4 dogs eventually required surgical intervention. Seven of 36 dogs in which otitis had resolved relapsed; 4 required additional lavage procedures. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that lavage of the tympanic bulla combined with medical management is an effective and viable option for treatment of otitis media in dogs. PMID- 15344363 TI - Use of the meridian test for the detection of equine herpesvirus type 1 infection in horses with decreased performance. AB - OBJECTIVE: To evaluate use of the acupuncture meridian test for detection of recent or recently reactivated equine herpesvirus type 1 (EHV-1) infection in horses with decreased performance. DESIGN: Case-control study. ANIMALS: 40 horses. PROCEDURE: Physical and neurologic examinations were performed, and acupuncture points on the bladder meridian were tested for sensitivity reactions in case and control horses. Polymerase chain reaction assays were performed to determine whether EHV-1 or equine herpesvirus type 4 (EHV-4) DNA could be detected in peripheral blood mononuclear cells. Complement fixation (CF) tests for detection of antibodies against EHV-1 and EHV-4 and virus neutralization (VN) tests for detection of antibodies against EHV-1 were performed on paired serum samples obtained 3 weeks apart. RESULTS: There was a significant difference in skin sensitivity in the cervical, sacral, and gluteal regions and flank between case and control horses. By use of the meridian test, all case horses were sensitive to manipulation of all acupuncture points believed to be associated with EHV infections, whereas only a few control horses were sensitive at an occasional point. Equine herpesvirus type 1 or EHV-4 viremia was not detected in any horses. Mean +/- SDVN antibody titers against EHV-1 were not significantly different between the 2 groups. Mean +/- SD CF antibody titers against EHV-1 obtained 3 weeks after the initial samples were higher in case horses than control horses; however, unequivocal seroconversion was not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the meridian test in case horses were associated with sensitivity reactions similar to those detected by physical and neurologic examinations; however, an unequivocal association with EHV-1 or EHV-4 infection was not detected. PMID- 15344364 TI - Effects of intravenous administration of dimethyl sulfoxide on cardiopulmonary and clinicopathologic variables in awake or halothane-anesthetized horses. AB - OBJECTIVE: To evaluate the cardiopulmonary and clinicopathologic effects of rapid IV administration of dimethyl sulfoxide (DMSO) in awake and halothane anesthetized horses. DESIGN: Prospective study. ANIMALS: 6 adult horses. PROCEDURES: Horses received IV infusion of 5 L of a balanced electrolyte solution with and without 1 g/kg (0.45 g/lb) of 10% DMSO solution when they were awake and anesthetized with halothane (4 treatments/horse). Arterial and venous blood samples were collected immediately before and at intervals during or after fluid administration and analyzed for blood gases and hematologic and serum biochemical variables, respectively. Heart rate, respiratory rate, and arterial blood pressure variables were recorded prior to, during, and after fluid administration. RESULTS: After administration of fluid with or without DMSO, changes in measured variables were detected immediately, but most variables returned to baseline values within 4 hours. One awake control horse had signs of anxiety; agitation and tachycardia were detected in 2 awake horses administered DMSO. These clinical signs disappeared when the rate of infusion was reduced. In anesthetized horses, increased concentrations of WBCs and plasma fibrinogen and serum creatine kinase activity persisted for 24 hours, which was related to the stress of anesthesia more than the effects of fluid administration. CONCLUSIONS AND CLINICAL RELEVANCE: Infusion of 5 L of balanced electrolyte solution with or without 10% DMSO induced minimal changes in cardiopulmonary function and clinicopathologic variables in either awake or halothane-anesthetized horses. Stress associated with anesthesia and recovery had a greater influence on measured variables in anesthetized horses than fluid administration. PMID- 15344365 TI - Prevalence of Salmonella spp on conventional and organic dairy farms. AB - OBJECTIVE: To describe the occurrence of fecal shedding, persistence of shedding over time, and serogroup classification of Salmonella spp on a large number of dairy farms of various sizes. DESIGN: Longitudinal study. SAMPLE POPULATION: 22,417 fecal samples from cattle and 4,570 samples from the farm environment on 110 organic and conventional dairy farms in Minnesota, Wisconsin, Michigan, and NewYork. PROCEDURE: 5 visits were made to each farm at 2-month intervals from August 2000 to October 2001. Fecal samples from healthy cows, calves, and other targeted cattle groups and samples from bulk tank milk, milk line filters, water, feed sources, and pen floors were collected at each visit. Bacterial culture was performed at 1 laboratory. RESULTS: Salmonella spp were isolated from 4.8% of fecal samples and 5.9% of environmental samples; 92.7% of farms had at least 1 Salmonella-positive sample. The 75th percentile for median within-herd prevalence of Salmonella spp in cattle for 5 sampling visits to a given farm was 2.0% and for maximum within-herd prevalence of Salmonella spp was 13.6%. Farms with a median within-herd prevalence of Salmonella spp of > or = 2.0% accounted for 76.3% of Salmonella-positive samples. There was no significant difference in the prevalence of Salmonella spp between conventional and organic farms. Seasonal differences in Salmonella shedding were observed. More farms had at least 1 serogroup B isolate than any other serogroup, whereas serogroup E1 was the most common among all Salmonella-positive samples. More than 1 serogroup was isolated on 76.4% of Salmonella-positive farms. CONCLUSIONS AND CLINICAL RELEVANCE: Salmonella spp were isolated from > 90% of dairy farms; however, 25% of farms accounted for > 75% of Salmonella-positive samples. This information is critical for the direction of intervention strategies to decrease the prevalence of Salmonella spp on dairy farms. PMID- 15344366 TI - Isolation of Salmonella spp from the environment of dairies without any history of clinical salmonellosis. AB - OBJECTIVE: To determine whether Salmonella spp could be isolated from the environment of free stall dairies in Wisconsin without any history of clinical salmonellosis and determine the serotype and antimicrobial susceptibility of any Salmonella isolates recovered from the environment. DESIGN: Cross-sectional study. STUDY POPULATION: 20 free stall dairies with no history of clinical salmonellosis. PROCEDURES: Dairy owners completed a questionnaire regarding management and production practices. Multiple swab samples were obtained from throughout the free stall facilities and submitted for bacterial culture for Salmonella spp. Odds ratios were calculated to compare herd-level risk factors between dairies from which Salmonella organisms were isolated and herds from which Salmonella organisms were not isolated. RESULTS: Salmonella organisms were isolated from 9 of the 20 (45%) dairies. Salmonella serotype Meleagridis was isolated from 4 dairies, S. Meleagridis and S. Kentucky were isolated from 2 dairies, S. Meleagridis and S. Cyprus were isolated from 1 dairy, S. Cerro was isolated from 1 dairy, and S. Corvallis was isolated from 1 dairy. All isolates were susceptible to all antimicrobial agents tested. None of the potential risk factors analyzed demonstrated a significant association with an increased likelihood of isolating Salmonella spp. CONCLUSIONS AND CLINICAL RELEVANCE: Environmental Salmonella contamination was demonstrated on free stall dairies with no history of clinical salmonellosis. PMID- 15344367 TI - Hemolysis and Heinz body formation associated with ingestion of red maple leaves in two alpacas. AB - Two adult female alpacas were evaluated for acute onset of weakness, pale mucous membranes, and signs of depression of unknown etiology. Both alpacas had intravascular hemolysis, anemia, and Heinz body formation and had been fed wilted red maple leaves. Clinical signs developed several days after ingestion of the leaves. No other toxin exposure was reported, and no parasites were detected. Dietary copper and nitrate-nitrite concentrations were evaluated and determined to be within reference limits. Both alpacas continued to become profoundly anemic with continued hemolysis. This necessitated blood transfusion as a life-saving intervention; however, prior to whole blood being available, transfusion with bovine hemoglobin glutamer was necessary. Both alpacas recovered without complications. PMID- 15344368 TI - Changing nursing education: creating our tipping point. PMID- 15344369 TI - Covering content and the additive curriculum: how can I use my time with students to best help them learn what they need to know? PMID- 15344370 TI - The professional socialization of graduating students in generic and two-plus-two baccalaureate completion nursing programs. AB - This non-experimental, descriptive study explored the differences in professional socialization between graduating students of two-plus-two baccalaureate completion nursing programs and generic baccalaureate nursing (BSN) programs. The survey population was graduating students from NLN-accredited baccalaureate nursing programs in a 15-state region. Autonomy, noted as a mark of professionalism, was measured by the Nursing Activity Scale developed by Schutzenhofer. No significant difference was found between the level of professional socialization of graduating students of two-plus-two baccalaureate completion nursing programs and generic BSN programs. No significant difference was found in professional socialization between students from the two basic RN education programs prior to BSN completion. A positive, but not significant, correlation was found between professional socialization and years of employment as an RN. No significant relationship was found between professional role socialization and area of major nursing experience for the baccalaureate completion nursing students. PMID- 15344371 TI - The competence of nursing graduates from problem-based programs in South Africa. AB - Although a significant body of research regarding problem-based learning (PBL) programs has been conducted during the past 2 decades, most of it relates to medical students and their curricula. There has also been very little research in the context of developing countries. In South Africa, most of the students who are admitted into nursing programs are from disadvantaged backgrounds, and it is important to assess the extent to which process-based curricula are appropriate for this group. The purpose of this study was to describe and evaluate the outcomes of PBL programs in nursing schools in South Africa in terms of the actual clinical practice and competence of graduates, and to compare these outcomes with those of graduates from conventional programs. The objectives of the study were to identify the characteristics of practice of graduates from PBL and conventional schools of nursing in South Africa, as described by the graduates and their supervisors, and secondly, to compare the practice characteristics of the two groups. PMID- 15344372 TI - Integrating bioterrorism education into nursing school curricula. AB - Due to the events of September 11, 2001 and the bioterrorism-related anthrax episodes, the United States has escalated efforts to better prepare the nation for terrorist attacks. Early recognition and management of a biological attack are largely dependent on the clinical expertise of frontline health care personnel. Nurses are recognized as an integral part of this team. Schools of nursing should integrate bioterrorism education into their curricula to address this growing frontier of health care management. This article outlines the necessary components of bioterrorism education for nurses, reviews examples of available resources to facilitate its inclusion, and suggests ways to integrate this material into nursing curricula. PMID- 15344373 TI - Using professional specialty competencies to guide course development. AB - This article describes how an RN-to-BSN community health nursing (CHN) course was reconceptualized from a traditional model to a competency-based model. The traditional course assigned students to CHN preceptors and required a set number of clinical contact hours. As clinical preceptor placement opportunities diminished, students and faculty became increasingly dissatisfied with the course structure and requirements. Faculty endorsed the use of professional competencies to measure course learning outcomes and selected competencies identified by the Association of Community Health Nursing Educators. These competencies were clustered into units, with learning activities and grading criteria based on the critical knowledge, values, and clinical skills needed to demonstrate mastery of specific competencies. Course faculty, rather than agency preceptors, assessed student learning outcomes and mastery of competencies. The students demonstrated mastery of competencies and liked the degree of self-directed learning that built on their professional status as RNs. PMID- 15344374 TI - Promoting integration of genetics core competencies into entry-level nursing curricula. AB - Nurse educators must respond to the growing need to teach genetics content in undergraduate nursing curricula. Recently developed genetics core competencies can be used to guide curriculum assessment and planning. This article describes a 5-year effort to integrate genetics education into a baccalaureate nursing curriculum and provides the results of a curriculum survey based on published genetics core competencies. PMID- 15344375 TI - A rapid and cost-effective screening method for identification of targeted embryonic stem cells. AB - Generation and characterization of knockout mice from targeted embryonic stem (ES) cells have become one of the most powerful approaches to study gene function in vivo. However, experiments to identify targeted ES clones can be both labor intensive and expensive when the targeting efficiency is low. Using the steroid receptor coactivator-3 (SRC-3) gene as a model, we have developed a rapid and cost-effective method for identification of targeted ES clones. Specifically, a promoterless LacZ sequence was fused to the 5'-coding sequence of SRC-3 in the targeting vector. After homologous recombination in ES cells, LacZ expression was regulated by the endogenous gene promoter. In cases of random insertions the beta galactosidase would more likely not be produced due to lacking promoter or missing amino acid-reading frame. Accordingly, targeted ES clones were enriched more than 30 times in the population of blue clones positive to X-gal staining. Therefore, targeted ES clones can be selected quickly and economically by X-gal staining in large scale followed by Southern blot analysis on small number of blue clones if the gene of interest is expressed in ES cells. This strategy is particularly useful when the targeting efficiency is low and a reporter such as LacZ or GFP for mouse gene expression is desired. PMID- 15344376 TI - Responses of membrane protection enzyme system of tobacco leaves on Hg, Cd and Pb stresses in soil. AB - Pot experiment was used to study the responses of membrane protection enzyme system of tobacco leaves on Hg, Cd and Pb stresses in soil. The results showed that POD activity gradually increased with increasing concetrations of Hg, Cd and Pb. CAT and SOD activity gradually decreased under three heavy metals common existing and SOD variation curve showed unimodal curve under single or two elements existing with increase of concentration of Hg, Cd and Pb. The effects of Hg, Cd and Pb in soil: three elemets together > two elements together > single element only. The effects resulted in an imbalance--activated oxygen produce and scavenge and physiological biochemical process disorder. There was a synergistic action for the effect of Hg, Cd and Pb in soil on membrane protection enzyme system in tobacco leaves. PMID- 15344377 TI - [Effects of HMBA on the expression of cell-cycle-associated genes in human hepatocarcinoma SMMC-7721 cells]. AB - In this study, the effects of HMBA on the expression of G0/G1 phase arrest associated genes in human hepatocarcinoma SMMC-7721 cells were investigated. Immunocytochemistry and in situ hybridization assay revealed that the levels of p21WAF1/CIP1 and p16 proteins and the level of p21WAF1/CIP1 mRNA were increased while the levels of CDK4 and Cyclin D1 proteins and c-myc mRNA were decreased in the cells treated with HMBA. These results showed that HMBA could up-regulate the expression of p21WAF1/CIP1 and p16 genes, down-regulate the activity of Cyclin D1 CDK4 and the transcription of c-myc gene which were necessary for cells entering into S phase, and so arrest the cells in G0/G1 phase, and induce the differentiation of human hepatocarcinoma SMMC-7721 cells. PMID- 15344378 TI - [The influence of temperature on the protein expression of human lung cancer cell line A549]. AB - The study on proteome of human cancer is helpful to explain its pathogenesis and make good effect on its prevention and cure. We compared the 2-DE maps of whole proteins of human lung cancer line A549 at 37 degrees C, 42 degrees C and 45 degrees C for the purpose of studying the expression of its heat shock proteins. Three temperature-sensitive differential spots were obtained and named as P1, P2, P3, respectively. Analyzed by MALDI-TOF-MS and Peptident software searched in the SWISS-PROT database, the three differential proteins were elementarily identified. P1 matches with two proteins belong to the Aldo-keto reductase family, P2 may be a new protein and P3 is Zinc finger protein 11A. PMID- 15344379 TI - The intracellular microenvironment in host cell-specific phosphorylation of SNT1 by the FGFR1 tyrosine kinase. AB - The phosphorylation of the fibroblast growth factor receptor (FGFR) kinase substrate SNT1 (also called FGFR substrate 2, FRS2) by FGFR tyrosine kinases is both host cell- and receptor isotype-specific. To study the determinants of the host cell-specific phosphorylation of SNT1 by FGFR1 tyrosine kinase, we constructed a chimeric receptor FGFR2IIIb/R1 that consisted of an FGFR2IIIb ligand-binding ectodomain and an FGFR1 tyrosine kinase domain. The chimeric FGFR2IIIb/R1 kinase mediated robust phosphorylation of SNT1 immediately after transfection in mouse 3T3 cells where the FGFR1 kinase was residential, and in proliferative aged prostate tumor epithelial cells (DTE-R1/100) that ectopically expressed FGFR1 kinase. This is in contrast to the fact that the robust phosphorylation of SNT1 by ectopic FGFR1 kinase is an acquisition property in DTE premalignant prostate epithelial cells, which normally do not express FGFR1. The data suggest that the microenvironment of intracellular, rather than components in the extracellular compartment, of host cells is important in permitting FGFR1 kinase to strongly phosphorylate SNT1. Together with our previous data that the acquisition of SNT1 phosphorylation activity is concurrent with the acquisition of mitogenic activity of FGFR1 in prostate epithelial cells, the results here further demonstrate that the phosphorylation of SNT1 is host cell specific and that alterations induced by chronic exposure to ectopic FGFR1 kinase are involved in acquisition of SNT1 phosphorylation activity to the ectopic FGFR1 in prostate epithelial cells. PMID- 15344380 TI - [Study on high efficiency tissue culture of mature seeds of rice (JiaHe ZaoZhan)]. AB - In order to improve the embryogenic callus induction rate and the regeneration rate of JiaHe-ZaoZhan rice, the influence of different factors were investigated, media with different hormones were used. Induction medium was supplemented with 1.5 mg/L 2,4-D + 3 mg/L NAA + 0.1 mg/L KT + 1 mg/L phytic acid + 20 mg/L PAA. Embryogenic call were treated under the condition of 25 degrees C before transferring to regeneration medium, the regeneration medium contained 0.5 mg/L 6 BA + 3 mg/L NAA + 0.5 mg/L KT + 1 mg/L phytic acid. The experiment results indicated that the hormone treatments had certain effects on the callus induction. Under the optimal medium, culture condition and the hormone treatments, the embryogenic callus induction could reach over 95%, and dry treatment of embryogenic callus had been found to increase the frequency of plant regeneration, significantly the plant regeneration rate could reach over 80%. Transplanted into pots, the young plants grew well. Then a experimental system with stability and high regenerating efficiency has been established for the mature seeds of rice (JiaHe-ZaoZhan). PMID- 15344381 TI - [Cloning and analyzing of the cDNA sequence of CHS-A gene of Narcissus]. AB - Chalcone synthase (CHS) is a key enzyme in the biosynthesis of all classes of flavonoids. The production of flower pigment is specifically regulated by the activity of CHS. We cloned the cDNA sequence of CHS-A gene from Narcissus by PCR and analyzed the coding sequence of gene. The result demonstrated that the sequence of the coding region was 1167bp, encoding a protein of 389 amino acid which was more than 80% homology with CHS of the other 8 plants, such as Nicotine abacus and Solana tuberosum. PMID- 15344382 TI - [Effect of tumor necrosis factor alpha and beta on ionizing radiation-induced apotosis]. AB - Present paper described the effect of human tumor necrosis factor alpha (hTNFalpha) and beta (hTNFbeta) on apoptosis induced by ionizing radiation in human embryonic lung cell line 2BS diploid cells and human lung cancer cell line A549 and SPC cells. The results showed that both hTNFalpha and hTNFbeta significantly inhibited 7Gy gamma-ray-induced apoptosis in 2BS cells. In contrast, hTNFalpha or hTNFbeta can increase the sensitivity of tumor cell lines (A549 and SPC cells) to radiation. Therefore, the results suggested that TNFalpha and TNFbeta had potential as a therapeutic agent to protect normal cell from the radiation-induced apoptosis and to sensitize the tumour cells for the damage effect of ionizing radiation. PMID- 15344383 TI - [Distribution of neuropeptide Y-like and beta-endorphin-like immunoreactivity in the nervous system and Hatschek's pit of amphioxus, Branchiostoma belcheri]. AB - Using immunohischemical method, we have localized for the first time in the amphioxus, Branchiostoma belcheri, neuropeptide Y (NPY)-like and beta-endorphin (beta-Ep)-like immunoreactivity in nervous system and Hatschek's pit. NPY immunoreactive (-ir) perikarya appeared in the front and middle areas of the telencephalon and midbrain, as well as in the hindbrain. Dense plexuses of NPY-ir fibers coursed with NPY-ir neurons and formed fine networks. NPY-ir perikarya and fibers were observed in the dorsal and middle areas of spinal cord. Beta-Ep-like immunoreactive perikarya and fibers were located in the front and middle areas of the midbrain, as well as in the spinal cord, but not in the telencephalon and hindbrain. The regions showing beta-Ep immunoreactivity were less than that of NPY. NPY-like and beta-Ep-like immunoreactivity were also found in the Hatschek's pit of amphioxus. These results suggest that NPY and beta-Ep, as an neurotransmitter as in fish, may be involved in the regulation of Hatschek's pit gonadotrophic cells secretory activity in amphioxus, and provide a new morphological evidence for the close relationship between brain and Hatschek's pit in amphioxus. PMID- 15344384 TI - [The inflammatory mediators induced endothelial cell injury through delay neutrophils apoptosis]. AB - This study aimed at elucidating the relationship between NTP apoptosis and endothelial cell injury. After isolation and purification, activated NTPs were incubated with endothelial cells directly or indirectly contact in a ratio of 5:1 and 10:1, meanwhile 10ng/ml IL-6 and 10% (v/v) burned serum added or not. Apoptosis and necrosis of endothelial cells were detected in situ. Activated NTP alone exerted no obviously detrimental effects on endothelial cells. However, when IL-6-or serum from burn patients was added, NTP apoptosis was delayed and endothelial cells were injured in the form of necrosis. Inflammatory mediators can delay NTP apoptosis, further research demonstrated that direct contact between the two types of cells was essential for NTP to impact injury on endothelial cells. PMID- 15344385 TI - [Immortalization of endothelial cells differentiated from mouse embryonic stem cells]. AB - This study is designed to immortalize endothelial cells differentiated from embryonic stem cells. The embryoid bodies (EB) formed in vitro from embryonic stem cells, were induced to differentiate into many "round cells" (the precursor of endothelial cells) by retinoic acid (RA) and transforming growth factor-beta1 (TGF-beta1). These "round cells" later formed the vascular tube-like structures. Studies by scanning electronic microscopy and light microscopy and immunocytochemistry, demonstrated that these tube-like structures were constituted by a large number of round and flat cells, which were positive for "vWF" and "CD34"staining. These results indicate they are vascular endothelial cells. To immortalize these cells, human telomerase reverse transcriptase (hTERT) cDNA was transfected into "round cells" by lipofectine. hTERT mRNA expression in transfected cells was confirmed by Dot blot, RT-PCR. Furthermore, 95% these transfected cells maintain the characteristic of endothelial cell, can proliferate in large quantity in vitro, and are able to form tubular structures. These results suggest that hTERT cDNA transfection can immortalize the induced endothelial cells and therefore may provide a new source of seed cells for vascular engineering. PMID- 15344386 TI - [Functional analysis of the alpha1G subunit of the T-type calcium channel in cellular proliferation]. AB - Increases of functional T-type calcium channel (T-channel) expression have been associated with cellular proliferation although evidence for this remains controversial. In the present study, we have used a variety of cellular, molecular and electrophysiological techniques to test the hypothesis that T-type channels play a causal role in the signaling pathway leading to proliferation. The results showed that stable over-expression of alpha1G T-channel subunit in HEK-293 cells conferred a significant growth advantage. Thus, cell population doubling time was reduced to 13.7 +/- 0.3 h in alpha1G transfectants, compared to control cultures (22.1 +/- 1.1 h) and flow cytometry analysis showed that this was due to a reduction in the number of alpha1G transfectants residing in the G0/G1 phases of the cell cycle compared to controls. The selective T-type calcium channel blocker, mibefradil, induced a dose-dependent inhibition of proliferation in alpha1G tansfectants. Furthermore, the Western blotting results proved that the level of protein expression of CDK2, cyclin A and cyclin E was high in alpha1G transfectants compared to control cultures. Our results demonstrate that the T-type calcium channel provides a significant growth advantage to HEK-293 cells that might occur via effects on the G1/S cell cycle mechanism. PMID- 15344387 TI - [Effects of different treatments on oocytes activation and parthenogenesis in mouse]. AB - In order to study effects of electro-fusion and strontium chloride (SrCl2) activation in nuclear transfer experiment on activation and development of mouse oocytes, concentration and treatment duration of SrCl2, electro-pulse and electro pulse combining SrCl2 were used to activate mouse oocytes which were obtained after hCG 17h. Activated oocytes were cultured in vitro in CZB medium. The results were as follows: 82.4% activation percentage was obtained when the oocytes were treated with 10mmol/L SrCl2 for 6h, it was significantly (P>0.05) higher than those obtained from that treated with the 5mmol/L or 10mmol/L SrCl2 for 4h. The activation percentage was not significantly different between 5mmol/L and 10mmol/L SrCl2 for 6h, but the percentage of morula and blastocyst in 10mmol/L SrCl2 6h group was significantly (P > 0.05) higher than those in 5mmol/L SrCl2 6h group. In the groups of treatment with electro-pulse, the best activation percentage (70.9%) was obtained when the oocytes were treated with 1.0kv/cm, 320micros, 3 pulses, but M + B percentage (7.9%) was low. In the groups of treatment with electro-pulse combining with SrCl2, the best result was acquired (activation and M + B percentage were 75.0% and 57.3% separately) when the oocytes were treated in 10mmol/L SrCl2 for 6h interval 1h after treated with 1.8kv/cm, 10s, 1pulse. These results show that the treatment with electro-pulse combining SrCl2 is a better way to mouse parthenogenesis. PMID- 15344388 TI - [Effects of different activation methods on in vitro development of bovine somatic cloned embryos]. AB - Effects of different activation methods on the cleavage and in vitro development of bovine somatic cloned embryos constructed by intracytoplasmic nuclear injection were compared. The results show that the cleavage and in vitro development rate were not different significantly for constructed embryos cultured in 6-DMAP comparing with those in 6-DMAP + cytochalasin B (CCB) after activation with Ionomycin. Culture duration (3 to 4 h) in 6-DMAP or 6-DMAP + CCB had no significant effects on the cleavage and in vitro development ability of reconstructed embryos. CCB addition in the activation medium was benefit to the development of constructed embryos, although the effect wasn't significant. Within 1 to 4 h, the longer interval duration of nuclear injection and reconstructed embryo activation was, the higher cleavage and the blastocyst development rate of reconstructed embryos were. PMID- 15344389 TI - [Repopulation with exogenous mitochondria in the cell]. AB - To create human mitochondrial DNA (mtDNA)-transferred cells as cell model of mitochondria defects-related diseases, platelet of normal young and old subjects were isolated as donor of mtDNA, then fused with mtDNA-depleted cells (rho0 cells) under induction of PEG1500. Auxtrophy test, cytochrome c oxidase activity assay and PCR amplification of mtDNA were done to confirm the transferring of mtDNA. Cell clones were visible in the medium 10 to 15 days after fusion, which grew well in medium lacking uridine, had positive COX activity and contained objective fragment of mtDNA by PCR, opposite to rho0 cells. Transferring of mtDNA to rho0 cells was identified, and mtDNA-transferred cell models were successfully created. PMID- 15344390 TI - [Maintenance of bactericidal activity of pectin stabilized aqueous solutions]. AB - Maintenance of the bactericidal activity and organoleptic properties of 3% and 5% stabilized aqueous solutions of pectin (Pepidol PEG, 3% and Pepidol PEG, 5%) stored for prolonged periods and at various temperatures was studied. Pepidol PEG was shown to preserve its bactericidal activity and organoleptic properties for 28 months when stored at rather wide ranges of the environmental temperature. The drug did not lose its properties after freezing and subsequent melting. The results of the study allowed to increase the period of Pepidol PEG use up to 18 months and the storage temperature ranges up to -30 degrees to +30 degrees C. PMID- 15344391 TI - [Molecular mechanisms of cephalosporin resistance in Gram-negative bacteria of the Enterobacteriaceae family]. AB - Extended spectrum beta lactamase genes were detected by the PCR in 87.6% of 231 Enterobacteriaceae strains isolated in medical institutions of Moscow, St. Petersburg, Tomsk and Nazran that showed a decrease in their susceptibility to 3rd generation cephalosporins. Alone or in various combinations TEM type beta lactamases were detected in 43.3% of the isolates, 46.8 and 51.2% of the isolates produced SHV type and CTX type beta lactamases respectively. Combinations of 2 and 3 different determinants were detected in 40 and 14% of the isolates respectively. Production of class C beta lactamases was suspected in 28% of the isolates by their resistance to cefoxitin. The gene of ACT type beta lactamase was detected in 1 strain of Klebsiella pneumoniae and the gene of CMY type beta lactamase was detected in 1 strain of Proteus mirabilis. By the NCCLS 100% of the isolates was susceptible to meropenem, 14% was susceptible to cefotaxime, 64% was susceptible to cefepime, 81% was susceptible to cefoperazone/sulbactam, 47% was susceptible to gentamicin, 57% was susceptible to amikacin and 36% was susceptible to ciprofloxacin. PMID- 15344392 TI - [Clinical efficacy of clarithromycin delayed release dosage form in the treatment of community-acquired pneumonia]. AB - The efficacy of clarithromycin retarded release dosage form in the treatment of patients with community-acquired pneumonia was estimated in a open prospective comparative study. The drug was administered in a dose of 500 mg once a day (29 patients) and in a dose of 500 mg twice a day (28 patients). The clinical efficacy in the compared regimens amounted to 90.6 and 93.3% of the patients respectively and the bacteriological efficacy was 93.3 and 84.6% respectively. The adverse reactions were observed in 15.6 and 30.0% of the cases respectively. PMID- 15344393 TI - [Therapy of bronchopulmonary infection in smokers]. AB - The efficacy and safety of Cifran OD, a new dosage form of ciprofloxacin with prolonged action were studied in the treatment of patients with bronchopulmonary infection (n=23). The drug was used orally in a dose of 1 g once a day. The patients were divided into 2 groups: group 1 included patients with aggravation of chronic bronchitis and group 2 included patients with bacterial pneumonia. The recovery was stated in 77.7% of the patients in group 1, the other 26.3% of the patients showed clinical improvement. In the patients with pneumonia (group 2) the recovery was recorded in 100% of the cases. No significant side effects were observed. PMID- 15344394 TI - [Efficacy of moxifloxacin (Avelox) in prophylaxis of infection in patients with profound neutropenia]. AB - Comparative efficacy of moxifloxacin and ciprofloxacin as prophylactics of infection in cancer patients with severe neutropenia after the chemotherapy was studied. The study included 40 patients with malignant lymphomas and solid tumore who received 52 courses of the aggressive chemotherapy. Twenty four patients (30 courses) received oral moxifloxacin in a dose of 400 mg once a day from the first day of the neutrophil count decrease below 1.0 x 10(9)/l until its recovery to > 1.0 x 10(9)/l or when the signs of infection appeared. In the control group 16 patients (22 courses) received oral ciprofloxacin in a dose of 500 mg twice a day. The patients in both the groups were compatible by the diagnosis, age and neutropenia duration. The median of the days of the febrile neutropenia duration in the patients prophylactically treated with moxifloxacin was statistically lower (2.1 vs 3.6 in the control group, p < 0.05). The incidence of febrile neutropenia in the moxifloxacin group was significantly lower than that in the control group (73 and 100% respectively, p = 0.01). The incidence of bacteriologically confirmed infection in the moxifloxacin group was also lower (6% vs 27.2%, p = 0.04). Therefore, moxifloxacin proved to be a more efficient agent vs ciprofloxacin (standard prophylactic) in prevention of febrile neutropenia and neutropenic infection in cancer patients, which is likely due to its higher activity against grampositive organisms. PMID- 15344395 TI - [Problems of etiotropic therapy of typhoid fever and approaches to their solution]. AB - The main problems of etiotropic therapy for typhoid fever lie in underestimate of the characteristic features of its pathogenesis and particularly in development of typhoid granulomas and their histogenesis, as well as in wide spread of typhoid fever pathogenic strains resistant to the routine chemotherapeutics, i.e. polyresistant strains. Some problems are due to incorrect choice of the antimicrobials and their combinations, optimal doses, administration routes and pathogenetic therapy. In the XXth centure an increase in the emergence and a change in the nature of the typhoid fever pathogen resistance to antimicrobials were observed. It was shown that from the pharmacologic and pharmacodynamic viewpoints the highest efficacy of typhoid fever therapy should be provided by the following antimicrobials: fluoroquinolones (except for norfloxacin), 3rd and 4th generation cephalosporins, aminopenicillins, chloramhenicol (levomycetin), combinations of 2nd and 3rd generation aminoglycosides with biseptol, aminopenicillins or doxycycline, as well as chloramphenicol combinations with aminopenicillins or 2nd to 4th generation cephalosporins. Practical recommendations for the etiotropic therapy of patients with typhoid fever during its outbreak or epidemic are presented. PMID- 15344396 TI - [An aromatic heptaene antibiotic levorin and its derivatives in muscle activity]. AB - The review is concerned with the outlooks for the use of levorin, a membrane active and channel forming polyene antibiotic, and its alkyl derivatives in muscle activity. In complex with cholesterol and ergosterol, the aromatic heptaene antibiotic levorin forms structural ionic channels of the molecular size in the lipid and cell membranes. Levorin increases the membrane permeability for monosucrose and other neutral molecules as follows: H2O > urea > acetamide > glycerine > ribose > arabinose > glucose > saccharose. As a channel forming compound, levorin is able to induce in the cell membranes of the muscle fibres formation of additional channels permeable for the cations and to increase the flow of the energy dependent substrates to the cells and the outburst of the metabolites from them during intensive muscle activity. Levorin several times decreases the surface tension of aqueous solutions. In some models of experimental animals levorin promoted an increase of the blood fluidity and accelerated the blood stream in the blood vessels both in rest and in muscle activity. Physical load in a high power zone increases the intensity of lipid peroxidation that results in fatigue and lower physical efficiency. Possible prevention of an increase of the rate of free radical reactions by levorin and its alkyl derivatives providing higher antioxidant protection is discussed. PMID- 15344397 TI - Investigation of influence of hydrogen gas on Pd/BST/Pt device by impedance spectroscopy. AB - A new application using barium strontium titanate (BST) thin films to fabricate the Pd/BST/Pt heterostructure for hydrogen gas detection has been investigated. To further understand the mechanism of this novel hydrogen sensing properties, impedance spectroscopy of the Pd/BST/Pt has been systematically studied and interpreted satisfactorily based on an equivalent circuit model. The alternate current (AC) electrical data of the extracted parameters and the properties of dielectric permittivity are discussed. The influence of hydrogen on the impedance spectra demonstrates that the charges induced by hydrogen ions mainly occur at the interface rather than in the bulk of the film. These experimental results provide clear evidence for the hydrogen sensing mechanism in the Pd/BST/Pt capacitive device. PMID- 15344398 TI - Flextensional ultrasonic motor using the contour mode of a square piezoelectric plate. AB - This paper presents the design, fabrication, and characterization of a new type of standing wave piezoelectric ultrasonic motor. The motor uses a metallic flextensional amplifier, or cymbal, to convert the contour mode vibrations of a square piezoelectric ceramic plate into flexural oscillations, which are further converted to produce rotary actuation by means of an elastic-fin friction drive. The motor operates on a single-phase electrical supply. A beryllium copper rotor design with three-fin configuration was adopted, and the geometry was varied to include different material thicknesses, fin lengths, and inclinations. The best stall torque and no load speed for a 25-mm square motor were 0.72 Nmm and 895 r/minute, respectively. The behavior of the stator structure was analyzed by ANSYS finite element software using harmonic and modal analyses. The vibration mode estimated by finite element modeling (FEM) was confirmed by laser Doppler vibration measurements. PMID- 15344399 TI - Experimental measurement and numerical analysis on resonant characteristics of piezoelectric disks with partial electrode designs. AB - Three experimental techniques are used in this study to access the influence of the electrode arrangement on the resonant characteristics of piezoceramic disks. These methods, including the amplitude-fluctuation electronic speckle pattern interferometry (AF-ESPI), laser Doppler vibrometer-dynamic signal analyzer (LDV DSA), and impedance analysis, are based on the measurement of full-field displacement, pointwise displacement, and electric impedance, respectively. In this study, one full electrode design and three nonsymmetrical partial electrode designs of piezoelectric disks are investigated. Because the clear fringe patterns measured by the AF-ESPI method will be shown only at resonant frequencies, both the resonant frequencies and the corresponding vibration mode shapes are successfully obtained at the same time for out-of-plane and in-plane motions. The second experimental method is the impedance analysis, which is used to measure the resonant and antiresonant frequencies. In addition to these experimental methods, LDV-DSA is used to determine the resonant frequencies of the vibration mode with out-of-plane motion. From the experimental results, the dependence of electrode design on the vibration frequencies and mode shapes is addressed. Numerical computations based on the finite element method are presented, and the results are compared with the experimental measurements. The effect of different designs of electrode is more significant in the in-plane modes than that in the out-of-plane modes. PMID- 15344400 TI - The noise conversion method for oscillatory systems. AB - The paper addresses a method for calculating the amplitude and phase power spectral density (PSD) functions of an oscillatory system (resonator, oscillator, bandpass filter, selective circuit, etc.) via the PSDs of its intrinsic noise sources and relevant transformation coefficients. A systematic description of the method is given for the scalar and vector noises. As an illustration, the noise transformation coefficients are derived for a piezoelectric series branch with fluctuating motional inductance, capacity, and losses, in which static capacity is disregarded. We then clarify the rules regarding the shaping of either PSD function. The importance of this method resides in the fact that it enables us to study particular finite ranges of the PSD function without using differential equations. PMID- 15344401 TI - On the flicker noise of ferrite circulators for ultra-stable oscillators. AB - The flicker noise of the ferrite circulator is a critical element in ultra-stable microwave oscillators, in which the signal reflected from the input of the reference cavity is exploited to stabilize the frequency. This paper explains why the circulator noise must be measured in isolation mode, proposes a measurement scheme, and provides experimental results. The observed flicker spans from -162 to -170 dB[rad2]/Hz at 1 Hz off the 9.2 GHz carrier, and at +19 dBm of input power. In the same conditions, the instrument limit is below -180 dB[rad2]/Hz. Experiments also give information on the mechanical stability of the microwave assembly, which is in the range of 10(-11) m. The measurement method can be used as the phase detector of a corrected oscillator; and, in the field of solid-state physics, it can be used for the measurement of random fluctuations in magnetic materials. PMID- 15344402 TI - A general form of perfectly matched layers for three-dimensional problems of acoustic scattering in lossless and lossy fluid media. AB - The concept of perfectly matched layer (PML) has been proven very effective in absorbing electromagnetic waves in lossless media. An extension of this method and a complete three-dimensional (3-D) scheme properly suited for finite difference, time-domain (FDTD) modeling of acoustic propagation and scattering in unbounded problems are presented in this paper. This generalized PML is constructed in such a way that it performs significant absorption of traveling waves in acoustics for both lossless and lossy media. Theoretically, no reflections occur when propagating waves encounter the lossy medium-PML interface, no matter what the angle of incidence is, introducing at the same time the possibility for further wave attenuation via the stretched coordinates idea. Numerical results support the suggested PML theory as well as reveal the proper modifications, which lead to the achievement of the optimum absorbing-boundary condition. PMID- 15344403 TI - Shear stress induced by a gas bubble pulsating in an ultrasonic field near a wall. AB - Some of the effects that therapeutic ultrasound has in medicine and biology may be associated with steady oscillations of gas bubbles in liquid, very close to tissue surface. The bubble oscillations induce on the surface steady shear stress attributed to microstreaming. A mathematical simulation of the problem for both free and capsulated bubbles, known as contrast agents, is presented here. The simulation is based on a solution of Laplace's equation for potential flow and existing models for microstreaming. The solution for potential flow was obtained numerically using a boundary integral method. The solution provides the evolution of the bubble shape, the distribution of the velocity potential on the surface, and the shear stress along the surface. The simulation shows that significant shear stresses develop on the surface when the bubble bounces near the tissue surface. In this case, pressure amplitude of 20 kPa generates maximal steady shear stress of several kilo Pascal. Substantial shear stress on the tissue surface takes place inside a circular zone with a radius about half of the bubble radius. The predicted shear stress is greater than stress that causes hemolysis in blood and several orders of magnitude greater than the physiological stress induced on the vessel wall by the flowing blood. PMID- 15344404 TI - An incremental learning algorithm with confidence estimation for automated identification of NDE signals. AB - An incremental learning algorithm is introduced for learning new information from additional data that may later become available, after a classifier has already been trained using a previously available database. The proposed algorithm is capable of incrementally learning new information without forgetting previously acquired knowledge and without requiring access to the original database, even when new data include examples of previously unseen classes. Scenarios requiring such a learning algorithm are encountered often in nondestructive evaluation (NDE) in which large volumes of data are collected in batches over a period of time, and new defect types may become available in subsequent databases. The algorithm, named Learn++, takes advantage of synergistic generalization performance of an ensemble of classifiers in which each classifier is trained with a strategically chosen subset of the training databases that subsequently become available. The ensemble of classifiers then is combined through a weighted majority voting procedure. Learn++ is independent of the specific classifier(s) comprising the ensemble, and hence may be used with any supervised learning algorithm. The voting procedure also allows Learn++ to estimate the confidence in its own decision. We present the algorithm and its promising results on two separate ultrasonic weld inspection applications. PMID- 15344405 TI - Synthesis of slanted and quasi-slanted SAW transducers. AB - This paper is devoted to the synthesis of interdigital transducers (IDTs) with variable electrode periodicity in the direction orthogonal to the SAW propagation, including straight electrodes (slanted or fan-shaped IDTs) and stepped electrodes (quasi-slanted IDTs). A polarity weighting of electrodes is the only tool for realization of high-frequency selectivity in fan-shaped (slanted) transducers. Quasi-slanted transducers (QST) offer additional opportunities: optimization of apertures, periods, and relative shifts of acoustic channels. Moreover, a discrete apodization gives an extra flexibility for QSTs in realization of desired frequency responses without increasing SAW diffraction sensitivity. Original synthesis algorithms have been developed for optimization of slanted and quasi-slanted IDTs in SAW filters with a wide range of characteristics. PMID- 15344406 TI - Design and fabrication of annular arrays for high-frequency ultrasound. AB - The design, fabrication, and performance of miniature high-frequency annular arrays are described. A 50-MHz, 2-mm-diameter, 7-element, equal-area annular array was fabricated and tested. The array elements were defined using photolithography and the electrical contacts were made using ultrasonic wire bonding. The resulting transducer produced pulses with a -6 dB bandwidth of 52% and an insertion loss of -16 dB. A radiation pattern was collected by scanning the transducer array above the tip of a glass fiber. A -6 dB two-way beam width of 75 microns was found at f/2. The radiation pattern decreased smoothly to less than -60 dB at a distance of 550 microns. PMID- 15344408 TI - Novel compact microstrip interdigital bandstop filters. PMID- 15344407 TI - Notch filters with novel microstrip, triangle-type resonators. PMID- 15344409 TI - Measured resonance characteristics of a 2-ghz-fundamental quartz resonator. PMID- 15344410 TI - Variational formulation of the equations for small fields superposed on finite biasing fields in an electroelastic body. PMID- 15344411 TI - Piezoelectromagnetic waves in a ceramic plate. PMID- 15344412 TI - Monitoring thermally-induced lesions with supersonic shear imaging. AB - Thermally-induced lesions are generally stiffer than surrounding tissues. We propose here to use the supersonic shear imaging technique (SSI) for monitoring high-intensity focused ultrasound (HIFU) therapy. This new elasticity imaging technique is based on remotely creating shear sources using an acoustic radiation force at different locations in the medium. In these experiments, an HIFU probe is used to generate lesions in fresh tissue samples. A diagnostic transducer, controlled by our ultrafast scanner, is located in the therapeutic probe focal plane. It is used for both generating the shear waves and imaging the resulting propagation at frame rates reaching 5,000 images/s. Movies of the shear wave propagation can be computed off-line. The therapeutic and imaging sequences are interleaved and a set of wave propagation movies is performed during the heating process. From each movie, elasticity estimations have been performed using an inversion algorithm. It demonstrates the feasibility of detecting and quantifying the hardness of HIFU-induced lesions using SSI. PMID- 15344413 TI - A digital ultrasonic system for small animal imaging. AB - In this paper, a 50 MHz digital ultrasonic imaging system designed for small animals and its applications to mouse embryos and tumors are presented. Utilizing the programmability of a fully digital system, several advanced imaging techniques are developed and implemented. A synthetic aperture focusing technique is implemented to increase the penetration and extend the depth of focus of a mechanical scan system using a high frequency single crystal transducer. An adaptive weighting technique is introduced to further reduce the sidelobes and enhance image contrast. Experimental results demonstrate efficacy of the proposed two-step adaptive focusing technique. For flow imaging, several flow estimation algorithms are evaluated. Simulations and flow phantom experiments show that the butterfly search technique offers superior flow imaging performance particularly when the SNR is low and the pulse bandwidth is high. The system is tested by imaging mouse embryos and microcirculation of a mouse tumor in vivo. It is demonstrated that the system is capable of simultaneously acquiring B-mode, color Doppler mode and power Doppler mode small animal images. The system set-up, adaptive focusing technique, flow estimation algorithms and imaging results are shown. PMID- 15344414 TI - Experimental studies of the thermal effects associated with radiation force imaging of soft tissue. AB - Many groups are studying acoustic radiation force-based imaging modalities to determine the mechanical properties of tissue. Acoustic Radiation Force Impulse (ARFI) imaging is one of these modalities that uses standard diagnostic ultrasound scanners to generate localized, impulsive, acoustic radiation force in tissue. This radiation force generates tissue displacements that are tracked using conventional correlation-based ultrasound methods. The dynamic response of tissue to this impulsive radiation force provides information about the mechanical properties of the tissue. The generation of micron-scale displacements using acoustic radiation force in tissue requires the use of high-intensity acoustic beams, and the soft tissue heating associated with these high-intensity beams must be evaluated to ensure safety when performing ARFI imaging in vivo. Experimental studies using thermocouples have validated Finite Element Method (FEM) models that simulate the heating of soft tissue during ARFI imaging. Spatial maps of heating measured with the thermocouples are in good agreement with FEM model predictions, with cooling time constants measured and modeled to be on the order of several seconds. Transducer heating during ARFI imaging has been measured to be less than 1 degrees C for current clinical implementations. These validated FEM models can now be used to simulate soft tissue heating associated with different transducers, beam spacing, focal configurations and thermal material properties. These experiments confirm that ARFI imaging of soft tissue is safe, although thermal response must be monitored when developing ARFI beam sequences for specific tissue types and organsystems. PMID- 15344415 TI - Two dimensional arrays for real time 3D intravascular ultrasound. AB - We have previously described 2D arrays operating at up to 10.0 MHz consisting of several thousand elements for transthoracic cardiac imaging and over a hundred elements for intracardiac imaging using 7 Fr to 12 Fr catheters. We have begun to explore forward viewing real time 3D phased array intravascular ultrasound, which may require imaging depths of a few centimeters to look down the axis of a vessel to view vulnerable atherosclerotic plaque. We used a noncoaxial based cable technology that allowed 100 signal wires to be placed inside a4.8 French IVUS lumen with an inner diameter of 1.3 mm. We pursued two different fabrication technologies for the building of the transducers. Each transducer was constructed in the forward viewing configuration to allow simultaneous real time B-scans, C scans and volumetric rendering of vessels and vascular stents distal to the catheter tip. In order to obtain the desired penetration depth, each transducer was constructed to operate at 10.0 MHz. The first method included an ordered array of 11 x 11 = 121 elements. In order to conform to the round aperture of the IVUS lumen, the comers were cut off, resulting in a total of 97 signal channels. Real time images include a 4 mm diameter vessel in a tissue mimicking phantom, an expanded stent and a stent in an excised sheep aorta. The second method is based upon a laser dicing technique that cuts the individual elements in a random pattern. This resulted in 61 signal channels. Real time 3D images of the AIUM test object were made with this transducer. PMID- 15344416 TI - Saw palmetto and benign prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is a common health issue that affects 8% of all men at the age of 40, 60% of men in their 70s, and 90% of those greater than 80 years of age. One-fourth of these men will develop moderate to severe lower urinary tract symptoms that greatly affect their quality of life. Recent evidence suggests that the use of saw palmetto leads to improvements in urinary function for those suffering from BPH. The favorable comparison of saw palmetto with tamsulosin, a well-known first line agent in the treatment of urinary tract symptoms, demonstrates promise towards a beneficial effect of this herbal agent, with very few, if any, adverse effects. However, what degree of this beneficial activity is due to placebo effects is yet to be determined. In addition, the precise mechanism of action of saw palmetto in men with BPH remains unclear. PMID- 15344417 TI - Antioxidant activity of extracts of peh-hue-juwa-chi-cao in a cell free system. AB - In traditional Chinese medicine preparations, Hedyotis diffusa (HD), Hedyotis corymbosa (HC) and Mollugo pentaphylla (MP) are often used interchangeably under the name of "Peh-Hue-Juwa-Chi-Cao (PHJCC)." Although studies have been conducted to characterize the therapeutic activities of these different plant species, their antioxidant activity has never been investigated. In this study, our aim was to evaluate the antioxidant and free radical scavenging activities of these three different plant materials. At a concentration of 10 mg/ml, results showed that HD possesses the strongest inhibition on the FeCl2-ascorbic acid induced lipid peroxidation in rat liver homogenate, followed by HC and MP. MP showed a weak anti-lipid peroxidation activity at 1 and 3 mg/ml. Using Electron Spin Resonance (ESR) analysis, the order of superoxide anion scavenging activity was HC > HD > MP. However, MP was found to have the greatest hydroxyl radical scavenging activity compared to HC and HD. In conclusion, all three species used as PHJCC in Taiwan exhibited different antioxidant and free radical scavenging activities; these differences could explain, at least in part, the variation in therapeutic properties of PHJCC products in the market. PMID- 15344418 TI - Effect of dang-gui-ji-hwang-yeum on osteoporosis in ovariectomized rats. AB - Dang-Gui-Ji-Hwang Yeum (DGJHY; an Oriental prescription), has been successfully used for the management of osteoporotic disorders in China, Japan and Korea. In this study, we have characterized the effect of DGJHY on osteoporosis-associated phenomena in ovariectomized (OVX) rats by measuring body weights and bone histomorphometries in sham, OVX and DGJHY-administered OVX rats. Light microscopic analyses showed a porous or an eroded appearance on the tibial trabecular bone surface in OVX rats, whereas those of the sham and DGJHY administered OVX rats were composed of fine particles. The trabecular bone area and the trabecular thickness in OVX rats were significantly lower than those of sham rats. Moreover, these reductions in OVX rats were significantly reversed by the administration of DGJHY for 7 weeks, and osteoclast numbers were also significantly reduced. Although no differences were observed between OVX and DGJHY-administered OVX rats and the sham animals at the T3 level, we have found significant differences between these two groups at the T4 level. However, serum phosphorus, calcium, mechanical strength and the surface appearance of osteoblasts in the DGJHY-administered OVX rats were similar to those of OVX rats. These results suggest that DGJHY is effective at preventing bone loss in OVX rats. PMID- 15344419 TI - Anti-inflammatory effect of bee venom on type II collagen-induced arthritis. AB - Bee venom (BV) has been used to relieve pain and reduce inflammation in traditional Oriental medicine, especially in chronic inflammatory diseases such as rheumatoid arthritis (RA). We previously reported that the BV injection into a traditional acupuncture point (Zusanli) reduced arthritis-associated edema and nociceptive responses in Freund's adjuvant-induced arthritis in rats (Kwon et al., 2001). This study was designed to evaluate the anti-inflammatory and anti cytokine effect of BV on a murine type-II collagen-induced arthritis (CIA) model. Male mice were immunized by spontaneous injection of 100 microg of an emulsion of bovine type-II collagen and complete Freund's adjuvant (CFA), with a booster injection after 2 weeks. In the experimental group, 0.1 ml BV was injected at acupuncture point (Zusanli) near both knees twice a week for a total of 5 times. In the control group, normal saline was injected at the same frequencies. These injections began 5 weeks after the first collagen injection. Starting the 3rd week after the first collagen injection, we examined limb swelling and severity of arthritis twice a week. At 8 weeks, mice were sacrificed and synovial tissue was examined with the light microscope and serum cytokines (IL-1beta and TNF alpha) were measured by ELISA. The incidence of arthritis, the mean arthritis index and the number of arthritic limbs were significantly lower in the treatment compared to the control group (63% versus 75%, 3.4% versus 8.5%, 23% versus 75%, respectively). Among the serum proinflammatory cytokines, the production of TNF alpha in the BV group was suppressed compared to the control group (59 +/- 4.5 versus 99.5 +/- 6.5, p < 0.05), but IL-1beta was not suppressed. The examination of the histopathology of the joints of murine CIA showed decreased inflammation signs and less lymphocyte infiltration after BV acupuncture therapy. Acupuncture therapy with BV suppressed the development of arthritis and caused inhibition of the immune responses in type-II collagen-induced arthritis. PMID- 15344420 TI - Inhibition of substrate-tumor cell adhesion under the effect of gnidilatimonoein purified from Daphne mucronata. AB - Alteration of the cell surface glycoproteins of cancerous cells correlate with malignancy potential. To evaluate the mediation of membrane glycoproteins in wehi 164 cancerous cells, under the effect of D. mucronata crude extract and one of its purified active components, gnidilatimonoein, their attachment to fibronectin coated wells were investigated. The plant extract, 27 microl/ml (equivalent to 0.54 mg/of plant leaves powder per ml of culture medium), as well as gnidilatimonoein (0.94 microM), were capable of quenching by 58% and 64%, respectively, the attachment of wehi-164 cells to fibronectin-coated wells (4 microg/ml). In addition to alteration of cell adhesive properties, the morphology of the treated cells were significantly changed upon treatment with the non-toxic dose of the plant extract or gnidilatimonoein. While the untreated cells have polygonal shapes, the treated cells had spherical shapes. PMID- 15344421 TI - 4-Acetyl-12,13-epoxyl-9-trichothecene-3, 15-diol from Isaria japonica mediates apoptosis of rat bladder carcinoma NBT-II cells by decreasing anti-apoptotic Bcl 2 expression and increasing pro-apoptotic Bax expression. AB - We studied the effect of 4-acetyl-12,13-epoxyl-9-trichothecene-3, 15-diol (AETD) isolated from Isaria japonica, one of the most popular Chinese fungal medicines, on the induction of apoptosis in rat bladder carcinoma NBT-II cells. AETD was cytotoxic to NBT-II cells, and this cytotoxic effect appears to be attributed to its induction of apoptotic cell death, as AETD induced nuclear morphological changes and internucleosomal DNA fragmentation, and increased the proportion of hypodiploid cells and activity of caspase-3. AETD treatment also decreased the expression of the anti-apoptotic protein Bcl-2 and increased the expression of the pro-apoptotic protein Bax. These results provide important information in understanding the mechanism(s) of AETD-induced apoptosis. PMID- 15344422 TI - Shiunko promotes epithelization of wounded skin. AB - Shiunko is a traditional botanic formula (ointment) which is used clinically for the treatment of wounded skin caused by cuts, abrasions, frost or burn. The aim of this study was to evaluate the effect of Shiunko on epithelization of wounded skin. Experimental cutting wounds on the back skin of Sprague-Dawley rats were induced. Different bacterial inoculations (Pseudomonus aeruginosa and Staphylococcus aureus) and treatment (Shiunko, Povidone-iodine and saline) were arranged herein. The incidences of infection and the speed of epithelization were evaluated. We observed that the incidences of wound infection following Pseudomonas aeruginosa inoculation were lower on both the Shiunko-treated group (0%, p < 0.01) and Povidine-iodine-treated group (5%, p < 0.05), than the saline treated group (40%). The Shiunko-treated group reported higher percentages of complete epithelization not only on the sterilized wounds (100%) but also on the contaminated wounds (90%) when compared to the saline-treated group (60% sterilized wounds, 40% and 50% contaminated wounds) on day 7 (p < 0.01). Povidone iodine did not promote epithelization of wounded skin, whereas Shiunko did. PMID- 15344423 TI - Neuroprotective effects of Hypericum perforatum on trauma induced by hydrogen peroxide in PC12 cells. AB - The standard extracts of Hypericum perforatum L. (SEHP), a well-known medicinal plant, are used for the treatment of depression, exhibited upgrading and significant protective effects on the trauma of PC12 cells induced by 200 microM H2O2 in a dose-dependent manner within 24-hour treatment. Cell viability was assessed by the MTT method, and in situ cellular hydrogen peroxide (H2O2)-induced oxidative stress was examined by measurement of reactive oxygen species (ROS) formation using CDCFH procedures. Intra- and extra-cellular ROS levels decreased significantly to 71.9% and 50.0% of the control at a moderate concentration of 20 microg/ml, respectively, suggesting that SEHP could easily enter the cells and play important roles in reducing ROS levels. Our results were proved by detection of DNA fragmentation and inspection of cell morphology of PC12 cells. SEHP can obviously block DNA fragmentation and prevent the cells from shrinking and turning round of H2O2-induced apoptosis in PC12 cells at concentrations of 10 approximately 100 microg/ml. This data suggests SEHP may be a candidate for application in neurodegenerative diseases such as Alzheimer's disease or Parkinson's disease. PMID- 15344424 TI - Acupressure and Transcutaneous Electrical Acupoint Stimulation in improving fatigue, sleep quality and depression in hemodialysis patients. AB - The purpose of this study was to test the effectiveness of acupressure and Transcutaneous Electrical Acupoint Stimulation (TEAS) on fatigue, sleep quality and depression in patients who were receiving routine hemodialysis treatment. The study was a randomized controlled trial; qualified patients were randomly assigned to acupressure, TEAS or control groups. Patients in the acupressure and TEAS groups received 15 minutes of treatment 3 times a week for 1 month, whereas patients in the control group only received routine unit care. A total of 106 patients participated in the study. Methods of measurement included the revised Piper Fatigue Scale (PFS), the Pittsburgh Sleep Quality Index and the Beck Depression Inventory. Data were collected at baseline, during the intervention and post-treatment. The results indicated that patients in the acupressure and TEAS groups had significantly lower levels of fatigue, a better sleep quality and less depressed moods compared with patients in the control group based upon the adjusted baseline differences. However, there were no differences between acupressure and TEAS groups in outcome measures. This study provides an alternative method for health care providers in managing dialysis patients with symptoms of fatigue, poor sleep or depression. PMID- 15344425 TI - A study of electrical conductance of meridian in the obese during weight reduction. AB - This study was designed to investigate the electrical conductance of meridian in the obese during weight reduction. Ten obese including overweight (Body Mass Index, BMI > 26) and 30 healthy (non-overweight) people were recruited from Chung Yuan Christian University. The obese subjects were instructed to follow a weight reduction program that included diet control, exercise and oral intake of Prozac. A device, the design of which is based on the Ryodoraku theorem, was used to measure the electrical conductance of 12 meridians on both sides of the subjects. The results showed that: (1) the average coefficient of variation of the electrical conductances in 24 meridians of the obese group was statistically different from that of the healthy group (p < 0.05); (2) the average coefficient of variation of electrical conductance in the obese after weight reduction was significantly decreased than before the weight reduction program (p < 0.05); (3) the BMI and the electrical conductance of meridian was correlated in the obese (r = -0.77, p < 0.001) as well as in the healthy group (r = -0.92, p < 0.001). These findings suggest that electrical conductance of meridians can be a parameter to monitor weight, especially for obese people. PMID- 15344426 TI - Preventive and therapeutic effects of acupuncture on bone mass in osteopenic ovariectomized rats. AB - This study was designed to (1) test the preventive and therapeutic effects of acupuncture on osteopenia in ovariectomized (OVX) rats, and (2) assess whether treatment of different acupuncture points causes different effects on tibiae, femora or lumbar spines. Thirty-five female Sprague-Dawley rats were divided into four groups: Sham (sham-operated, non-acupuncture); Model (OVX, non-acupuncture); Acp-A [OVX, bilateral needling of points Tsu-San-Li (ST-36) and San-Yin- Chiao (SP-6)]; and Acp-B (OVX, bilateral needling of P'i-Shu (Bl-20) and Shen Shu (Bl 23)). Operations were performed at 8 weeks of age, 1 week later the study was started and continued for 16 weeks. Ovariectomy resulted in decreased bone mineral density (BMD) compared to the sham group over time, and Acp-A tended to have higher BMD than the other OVX groups, especially for tibiae. In addition, the bone ash weight of the Acp groups tended to be heavier than the model group. Deoxypyridinoline, the urinary marker of bone resorption, also appeared to be decreased in both acupuncture groups. Similarly, microarchitecture and bone morphometry of lumbar vertebrae and tibiae, such as bone volume, trabecular thickness, trabecular number, mineralizing surface, bone formation rate, mineral apposition rate, number of nodes and number of node-terminus struts, also showed the same improvement in the acupuncture groups as compared to the model control group. Our findings showed that acupuncture may prevent the development of osteopenia in rats induced by ovariectomy. Needling of Tsu-San-Li (ST-36) and San Yin- Chiao (SP-6) seems more effective than needling of P'i-Shu (Bl-20) and Shen Shu (Bl-23) in bone anabolic regulation. PMID- 15344427 TI - The effectiveness of auriculoacupoint treatment for artificially induced acute hepatic injury in dogs. AB - This study was to investigate effects of auriculoacupoint (AAP) treatment on carbon tetrachloride (CCl4)-induced acute hepatic injury in ten dogs (four females and six males). Dogs have been divided into two groups: the control group (four dogs), not stimulated after induction of hepatic injury and the experimental group (six dogs), stimulated with AAP and massaged at the affected liver region of internal ear after induction of hepatic injury. Serum enzyme activities and histopathological findings were examined after the application of AAP. Compared to the control group, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in the experimental group were significantly decreased at the 4th day (p < 0.05) and at the 5th day (p < 0.05), respectively. Histopathological findings of the experimental group showed decrease of necrotic region and size of lipid droplets compared to the control group. In conclusion, AAP treatment had a therapeutic effect on the recovery of liver injury induced by CCl4 in dogs. PMID- 15344428 TI - Effects of Tai Chi exercise on physical and mental health of college students. AB - The purpose of this pilot study is to examine the effects of Tai Chi Quan, a body mind harmony exercise, on college students' perceptions of their physical and mental health. A three-month intervention of Tai Chi exercise was administered to college students, and multidimensional physical (PHD) and mental (MHD) health scores were assessed using the SF-36v2 health survey questionnaire before and after the intervention. Thirty college students participated in a 1-hour-long Tai Chi exercise intervention twice a week for 3 months. Each practice session included 10 minutes of breathing and stretching exercises followed by 50 minutes of Tai Chi Quan 24-form practice. PHD including physical function (PF), role physical (RP), bodily pain (BP), general health (GH), and MHD including social function (SF), role mental/emotion function (RE), vitality (VT), perceptions of mental health (MH) were assessed. The normalized scores of each variable and the combined PHD or MHD scores before and after the Tai Chi intervention were examined by paired t-test (p < 0.05). Physical measures of BP and GH, and mental measures of RE, VT and MH were significantly improved after Tai Chi exercise intervention. When the overall PHD or MHD scores were evaluated, the MHD had increased significantly. In conclusion, Tai Chi exercise had positive effects on the self-assessed physical and mental health of college students. Scores on the mental health dimension appeared to be particularly sensitive to change. Colleges/universities might consider offering Tai Chi as a component of their ongoing physical activity programs available to students. PMID- 15344429 TI - External Qi therapy to treat symptoms of Agent Orange Sequelae in Korean combat veterans of the Vietnam War. AB - We investigated the efficacy of Qi therapy as a non-pharmacological treatment for various symptoms presented by Korean combat veterans of the Vietnam War with Agent Orange Sequelae. Nine subjects volunteered to receive 30 minutes of Qi therapy, twice per day for 7 days. There was marked improvement in 89% of the patients with impaired physical activity, 86% of those with psychological disorder, 78% of those with heavy drug use, and 67% of those with fatigue, indigestion and high blood glucose levels. This data suggests that Qi therapy combined with conventional treatment has positive effects in reducing and managing the pain, psychosomatic disorders, and substance abuse in patients with Agent Orange Sequelae. We cannot completely discount the possible influence of the placebo effect, and more objective, clinical measures are needed to study the long-term effects of Qi therapy. PMID- 15344430 TI - A culturally sensitive health care practice model--theory construction and its testing. AB - The major purpose of this paper was to develop a model for the use of complementary and alternative health care options. Chinese-American families were selected due to their large proportion among all ethnic populations in the United States. Andersen's (1973 and 1995) perception of predisposing and enabling was adopted to test study variables singly and interactively on their health care use. The dependent variable was defined as health behavior preference towards Chinese treatment (also known as complementary and alternative medicine treatment) or Western treatment, and a combination of Chinese and Western treatment (also known as integrative medicine treatment). Data were randomly collected from 240 adult Chinese-American respondents out of 1080 members of the Chinese American Association in the greater Cleveland area. Findings both in the full and partial research models reveal that the factors contributing the most to their medical preferences were their acculturation level and health beliefs. Inter-generational differences in health care use were also discussed. PMID- 15344431 TI - Professional hallmarks. PMID- 15344432 TI - Standard of care? PMID- 15344433 TI - Assessing 'real science'. PMID- 15344434 TI - Teacher motivates dental professionals with 'real life' story. PMID- 15344435 TI - New advances in science and technology of endodontics. PMID- 15344436 TI - Microbiologic aspects of endodontic infections. AB - Our understanding of endodontic infections and treatment of endodontic disease has increased significantly over the last decade. This article is an update of those findings. Aspects that are reviewed include: portal of entry for microorganisms, virulence and pathogenicity of organisms, descriptions of primary and recurrent endodontic infections, and treatment of endodontic infections. PMID- 15344437 TI - New advances in the management of endodontic pain emergencies. AB - The development of more effective strategies for managing endodontic pain emergencies draws from the research of both clinicians and basic scientists. This review explores evidence-based approaches for managing endodontic pain emergencies and newly emerging pain management strategies based upon molecular, cellular and physiologic research into pain mechanisms. PMID- 15344438 TI - Nonsurgical endodontic retreatment. AB - The purpose of this clinical article is to emphasize that although there is enormous potential for endodontic success, clinicians are, at times, confronted with post-treatment disease. A rationale for endodontic treatment is followed by the goals of nonsurgical retreatment. The focus of this article is to identify the various nonsurgical retreatment categories and provide an overview of the concepts, armamentarium and techniques available to disassemble roots, address deficiencies or repair defects that are pathological or iatrogenic in origin. PMID- 15344439 TI - Contemporary endodontic surgery. AB - During the past decade, endodontics has seen a dramatic shift in the application of periradicular surgery and the role it plays in endodontic treatment. With the introduction of enhanced magnification, periradicular ultrasonics and other associative technologies, teeth that might otherwise be extracted now have a chance for retention. This article describes the role of these advances in contemporary endodontic surgery. PMID- 15344440 TI - The success of endodontic therapy--healing and functionality. AB - Current, relevant knowledge on the outcome of endodontic therapy is key to clinical decision making, particularly when endodontic treatment is weighed against tooth extraction and replacement. Inherent to reviewing the outcome is a definition of "success" in relation to the goals of therapy. As the specific goal set out by the individual patient may either be healing/prevention of disease (apical periodontitis) or just functional retention of the tooth, the potential for both healing and functionality is reviewed. Based on selected follow-up studies that offer the best evidence, the chance of teeth without apical periodontitis to remain free of disease after initial treatment or orthograde retreatment is 92 percent to 98 percent. The chance of teeth with apical periodontitis to completely heal after initial treatment or retreatment is 74 percent to 86 percent, and their chance to be functional over time is 91 percent to 97 percent. Thus there does not appear to be a systematic difference in outcome between initial treatment and orthograde retreatment. The outcome of apical surgery is less consistent than that of the nonsurgical treatment. The chance of teeth with apical periodontitis to completely heal after apical surgery is 37 percent to 85 percent, with a weighted average of approximately 70 percent. However, even with the lower chance of complete healing, the chance for the teeth to be functional over time is 86 percent to 92 percent. Considering the favorable outcome, conservative endodontic therapy, both nonsurgical and surgical, is definitely justified and should be attempted when a good restorative and periodontal prognosis is projected, unless the patient is not motivated to retain the tooth. PMID- 15344441 TI - Fun with gold foil. PMID- 15344442 TI - Shape modelling using Markov random field restoration of point correspondences. AB - A method for building statistical point distribution models is proposed. The novelty in this paper is the adaption of Markov random field regularization of the correspondence field over the set of shapes. The new approach leads to a generative model that produces highly homogeneous polygonized shapes and improves the capability of reconstruction of the training data. Furthermore, the method leads to an overall reduction in the total variance of the point distribution model. Thus, it finds correspondence between semi-landmarks that are highly correlated in the shape tangent space. The method is demonstrated on a set of human ear canals extracted from 3D-laser scans. PMID- 15344443 TI - Optimal deformable surface models for 3D medical image analysis. AB - We present a novel method for building an optimal statistical deformable model from a set of surfaces whose topological realization is homeomorphic to a compact 2D manifold with boundary. The optimal parameterization of each shape is recursively refined by using hierarchical PBMs and tensor product B-spline representation of the surface. A criterion based on MDL is used to define the internal correspondence of the training data. The strength of the proposed technique is demonstrated by deriving a concise statistical model of the human left ventricle which has principal modes of variation that correspond to intrinsic cardiac motions. We demonstrate how the derived model can be used for 3D dynamic volume segmentation of the left ventricle, with its accuracy assessed by comparing results obtained from manual delineation of 3D cine MR data of 8 asymptomatic subjects. The extension of the technique to shapes with complex topology is also discussed. PMID- 15344444 TI - Learning object correspondences with the observed transport shape measure. AB - We propose a learning method which introduces explicit knowledge to the object correspondence problem. Our approach uses an a priori learning set to compute a dense correspondence field between two objects, where the characteristics of the field bear close resemblance to those in the learning set. We introduce a new local shape measure we call the "observed transport measure", whose properties make it particularly amenable to the matching problem. From the values of our measure obtained at every point of the objects to be matched, we compute a distance matrix which embeds the correspondence problem in a highly expressive and redundant construct and facilitates its manipulation. We present two learning strategies that rely on the distance matrix and discuss their applications to the matching of a variety of 1-D, 2-D and 3-D objects, including the corpus callosum and ventricular surfaces. PMID- 15344445 TI - Shape discrimination in the hippocampus using an MDL model. AB - We extend recent work on building 3D statistical shape models, automatically, from sets of training shapes and describe an application in shape analysis. Using an existing measure of model quality, based on a minimum description length criterion, and an existing method of surface re-parameterisation, we introduce a new approach to model optimisation that is scalable, more accurate, and involves fewer parameters than previous methods. We use the new approach to build a model of the right hippocampus, using a training set of 82 shapes, manually segmented from 3D MR images of the brain. We compare the results with those obtained using another previously published method for building 3D models, and show that our approach results in a model that is significantly more specific, general, and compact. The two models are used to investigate the hypothesis that there are differences in hippocampal shape between age-matched schizophrenic and normal control subgroups within the training set. Linear discriminant analysis is used to find the combination of shape parameters that best separates the two subgroups. We perform an unbiased test that shows there is a statistically significant shape difference using either shape model, but that the difference is more significant using the model built using our approach. We show also that the difference between the two subgroups can be visualised as a mode of shape variation. PMID- 15344446 TI - Minimum Description Length shape and appearance models. AB - The Minimum Description Length (MDL) approach to shape modelling is reviewed. It solves the point correspondence problem of selecting points on shapes defined as curves so that the points correspond across a data set. An efficient numerical implementation is presented and made available as open source Matlab code. The problems with the early MDL approaches are discussed. Finally the MDL approach is extended to an MDL Appearance Model, which is proposed as a means to perform unsupervised image segmentation. PMID- 15344447 TI - Evaluation of 3D correspondence methods for model building. AB - The correspondence problem is of high relevance in the construction and use of statistical models. Statistical models are used for a variety of medical application, e.g. segmentation, registration and shape analysis. In this paper, we present comparative studies in three anatomical structures of four different correspondence establishing methods. The goal in all of the presented studies is a model-based application. We have analyzed both the direct correspondence via manually selected landmarks as well as the properties of the model implied by the correspondences, in regard to compactness, generalization and specificity. The studied methods include a manually initialized subdivision surface (MSS) method and three automatic methods that optimize the object parameterization: SPHARM, MDL and the covariance determinant (DetCov) method. In all studies, DetCov and MDL showed very similar results. The model properties of DetCov and MDL were better than SPHARM and MSS. The results suggest that for modeling purposes the best of the studied correspondence method are MDL and DetCov. PMID- 15344448 TI - Localization of anatomical point landmarks in 3D medical images by fitting 3D parametric intensity models. AB - We introduce a new approach for the localization of 3D anatomical point landmarks based on 3D parametric intensity models which are directly fit to the image. We propose an analytic intensity model based on the Gaussian error function in conjunction with 3D rigid transformations as well as deformations to efficiently model tip-like structures of ellipsoidal shape. The approach has been successfully applied to accurately localize anatomical landmarks in 3D MR and 3D CT image data. We have also compared the experimental results with the results of a previously proposed 3D differential operator. It turns out that the new approach significantly improves the localization accuracy. PMID- 15344449 TI - Morphology-based cortical thickness estimation. AB - We describe a new approach to estimating the cortical thickness of human brains using magnetic resonance imaging data. Our algorithm is part of a processing chain consisting of a brain segmentation (skull stripping), as well as white and grey matter segmentation procedures. In this paper, only the grey matter segmentation together with the cortical thickness estimation is described. In contrast to many existing methods, our estimation method is voxel-based and does not use any surface meshes. While this fact poses a principal limit on the accuracy that can be achieved by our method, it offers tremendous advantages with respect to practical applicability. In particular, it is applicable to data sets showing severe cortical atrophies that involve areas of high curvature and extremely thin gyral stalks. In contrast to many other methods, it is entirely automatic and very fast with computation times of a few minutes. Our method has been used in two clinical studies involving a total of 27 patients and 23 healthy subjects. PMID- 15344450 TI - The shape operator for differential analysis of images. AB - This work provides a new technique for surface oriented volumetric image analysis. The method makes no assumptions about topology, instead constructing a local neighborhood from image information, such as a segmentation or edge map, to define a surface patch. Neighborhood constructions using extrinsic and intrinsic distances are given. This representation allows one to estimate differential properties directly from the image's Gauss map. We develop a novel technique for this purpose which estimates the shape operator and yields both principal directions and curvatures. Only first derivatives need be estimated, making the method numerically stable. We show the use of these measures for multi-scale classification of image structure by the mean and Gaussian curvatures. Finally, we propose to register image volumes by surface curvature. This is particularly useful when geometry is the only variable. To illustrate this, we register binary segmented data by surface curvature, both rigidly and non-rigidly. A novel variant of Demons registration, extensible for use with differentiable similarity metrics, is also applied for deformable curvature-driven registration of medical images. PMID- 15344451 TI - Feature selection for shape-based classification of biological objects. AB - This paper introduces a method for selecting subsets of relevant statistical features in biological shape-based classification problems. The method builds upon existing feature selection methodology by introducing a heuristic that favors the geometric locality of the selected features. This heuristic effectively reduces the combinatorial search space of the feature selection problem. The new method is tested on synthetic data and on clinical data from a study of hippocampal shape in schizophrenia. Results on clinical data indicate that features describing the head of the right hippocampus are most relevant for discrimination. PMID- 15344452 TI - Corresponding articular cartilage thickness measurements in the knee joint by modelling the underlying bone (commercial in confidence). AB - We present a method for corresponding and combining cartilage thickness readings from a population of patients using the underlying bone structure as a reference. Knee joint femoral bone and cartilage surfaces are constructed from a set of parallel slice segmentations of MR scans. Correspondence points across a population of bone surfaces are defined and refined by minimising an objective function based on the Minimum Description Length of the resulting statistical shape model. The optimised bone model defines a set of corresponding locations from which 3D measurements of the cartilage thickness can be taken and combined for a population of patients. Results are presented for a small group of patients demonstrating the feasibility and potential of the approach as a means of detecting sub-millimetre cartilage thickness changes due to disease progression. PMID- 15344453 TI - Adapting Active Shape Models for 3D segmentation of tubular structures in medical images. AB - Active Shape Models (ASM) have proven to be an effective approach for image segmentation. In some applications, however, the linear model of gray level appearance around a contour that is used in ASM is not sufficient for accurate boundary localization. Furthermore, the statistical shape model may be too restricted if the training set is limited. This paper describes modifications to both the shape and the appearance model of the original ASM formulation. Shape model flexibility is increased, for tubular objects, by modeling the axis deformation independent of the cross-sectional deformation, and by adding supplementary cylindrical deformation modes. Furthermore, a novel appearance modeling scheme that effectively deals with a highly varying background is developed. In contrast with the conventional ASM approach, the new appearance model is trained on both boundary and non-boundary points, and the probability that a given point belongs to the boundary is estimated non-parametrically. The methods are evaluated on the complex task of segmenting thrombus in abdominal aortic aneurysms (AAA). Shape approximation errors were successfully reduced using the two shape model extensions. Segmentation using the new appearance model significantly outperformed the original ASM scheme; average volume errors are 5.1% and 45% respectively. PMID- 15344454 TI - A unified variational approach to denoising and bias correction in MR. AB - We propose a novel bias correction method for magnetic resonance (MR) imaging that uses complementary body coil and surface coil images. The former are spatially homogeneous but have low signal intensity; the latter provide excellent signal response but have large bias fields. We present a variational framework where we optimize an energy functional to estimate the bias field and the underlying image using both observed images. The energy functional contains smoothness-enforcing regularization for both the image and the bias field. We present extensions of our basic framework to a variety of imaging protocols. We solve the optimization problem using a computationally efficient numerical algorithm based on coordinate descent, preconditioned conjugate gradient, half quadratic regularization, and multigrid techniques. We show qualitative and quantitative results demonstrating the effectiveness of the proposed method in producing debiased and denoised MR images. PMID- 15344455 TI - Object-based strategy for morphometry of the cerebral cortex. AB - Most of the approaches dedicated to automatic morphometry rely on a point-by point strategy based on warping each brain towards a reference coordinate system. In this paper, we describe an alternative object-based strategy dedicated to the cortex. This strategy relies on an artificial neuroanatomist performing automatic recognition of the main cortical sulci and parcellation of the cortical surface into gyral patches. A set of shape descriptors, which can be compared across subjects, is then attached to the sulcus and gyrus related objects segmented by this process. The framework is used to perform a study of 142 brains of the ICBM database. This study reveals some correlates of handedness on the size of the sulci located in motor areas, which seem to be beyond the scope of the standard voxel based morphometry. PMID- 15344456 TI - Genus zero surface conformal mapping and its application to brain surface mapping. AB - It is well known that any genus zero surface can be mapped conformally onto the sphere and any local portion thereof onto a disk. However, it is not trivial to find a general method which finds a conformal mapping between two general genus zero surfaces. We propose a new variational method which can find a unique mapping between any two genus zero manifolds by minimizing the harmonic energy of the map. We demonstrate the feasibility of our algorithm by applying it to the cortical surface matching problem. We use a mesh structure to represent the brain surface. Further constraints are added to ensure that the conformal map is unique. Empirical tests on MRI data show that the mappings preserve angular relationships, are stable in MRIs acquired at different times, and are robust to differences in data triangulation, and resolution. Compared with other brain surface conformal mapping algorithms, our algorithm is more stable and has good extensibility. PMID- 15344457 TI - Coupled multi-shape model and mutual information for medical image segmentation. AB - This paper presents extensions which improve the performance of the shape-based deformable active contour model presented earlier in [9]. In contrast to that work, the segmentation framework that we present in this paper allows multiple shapes to be segmented simultaneously in a seamless fashion. To achieve this, multiple signed distance functions are employed as the implicit representations of the multiple shape classes within the image. A parametric model for this new representation is derived by applying principal component analysis to the collection of these multiple signed distance functions. By deriving a parametric model in this manner, we obtain a coupling between the multiple shapes within the image and hence effectively capture the co-variations among the different shapes. The parameters of the multi-shape model are then calculated to minimize a single mutual information-based cost functional for image segmentation. The use of a single cost criterion further enhances the coupling between the multiple shapes as the deformation of any given shape depends, at all times, upon every other shape, regardless of their proximity. We demonstrate the utility of this algorithm to the segmentation of the prostate gland, the rectum, and the internal obturator muscles for MR-guided prostate brachytherapy. PMID- 15344458 TI - Neighbor-constrained segmentation with 3D deformable models. AB - A novel method for the segmentation of multiple objects from 3D medical images using inter-object constraints is presented. Our method is motivated by the observation that neighboring structures have consistent locations and shapes that provide configurations and context that aid in segmentation. We define a Maximum A Posteriori(MAP) estimation framework using the constraining information provided by neighboring objects to segment several objects simultaneously. We introduce a representation for the joint density function of the neighbor objects, and define joint probability distributions over the variations of the neighboring positions and shapes of a set of training images. By estimating the MAP shapes of the objects, we formulate the model in terms of level set functions, and compute the associated Euler-Lagrange equations. The contours evolve both according to the neighbor prior information and the image gray level information. We feel that this method is useful in situations where there is limited inter-object information as opposed to robust global atlases. Results and validation from various experiments on synthetic data and medical imagery in 2D and 3D are demonstrated. PMID- 15344459 TI - Expectation maximization strategies for multi-atlas multi-label segmentation. AB - It is well-known in the pattern recognition community that the accuracy of classifications obtained by combining decisions made by independent classifiers can be substantially higher that the accuracy of the individual classifiers. In order to combine multiple segmentations we introduce two extensions to an expectation maximization (EM) algorithm for ground truth estimation based on multiple experts (Warfield et al., MICCAI 2002). The first method repeatedly applies the Warfield algorithm with a subsequent integration step. The second method is a multi-label extension of the Warfield algorithm. Both extensions integrate multiple segmentations into one that is closer to the unknown ground truth than the individual segmentations. In atlas-based image segmentation, multiple classifiers arise naturally by applying different registration methods to the same atlas, or the same registration method to different atlases, or both. We perform a validation study designed to quantify the success of classifier combination methods in atlas-based segmentation. By applying random deformations, a given ground truth atlas is transformed into multiple segmentations that could result from imperfect registrations of an image to multiple atlas images. We demonstrate that a segmentation produced by combining multiple individual registration-based segmentations is more accurate for the two EM methods we propose than for simple label averaging. PMID- 15344460 TI - Quantitative analysis of intrathoracic airway trees: methods and validation. AB - A method for quantitative assessment of tree structures is reported allowing evaluation of airway or vascular tree morphology and its associated function. Our skeletonization and branch-point identification method provides a basis for tree quantification or tree matching, tree-branch diameter measurement in any orientation, and labeling individual branch segments. All main components of our method were specifically developed to deal with imaging artifacts typically present in volumetric medical image data. The proposed method has been tested in 343 computer phantom instances subjected to changes of its orientation as well as in a repeatedly CT-scanned rubber plastic phantom width sub-voxel accuracy and high reproducibility. Application to 35 human in vivo trees yielded reliable and well-positioned centerlines and branch-points. PMID- 15344461 TI - Multi-View Active Appearance Models: application to X-ray LV angiography and cardiac MRI. AB - This paper describes a Multi-View Active Appearance Model (AAM) for coherent segmentation of multiple cardiac views. Cootes' AAM framework was adapted by considering shapes and intensities from multiple views, while eliminating trivial difference in object pose in different views. This way, the coherence in organ shape and intensities between different views is modeled, and utilized during image search. The method is validated in two substantially different and novel applications: segmentation of combined end-diastolic and end-systolic left ventricular X-ray angiograms, and simultaneous segmentation of a combination of four chamber, two chamber and short-axis cardiac MR views. PMID- 15344462 TI - Tunnelling descent: a new algorithm for active contour segmentation of ultrasound images. AB - The presence of speckle in ultrasound images makes it hard to segment them using active contours. Speckle causes the energy function of the active contours to have many local minima, and the gradient descent procedure used for evolving the contour gets trapped in these minima. A new algorithm, called tunnelling descent, is proposed in this paper for evolving active contours. Tunnelling descent can jump out of many of the local minima that gradient descent gets trapped in. Experimental results with 70 short axis cardiac ultrasound images show that tunnelling descent has no trouble finding the blood-tissue boundary (the endocardium). This holds irrespective of whether tunnelling descent is initialized in blood or tissue. PMID- 15344463 TI - Improving appearance model matching using local image structure. AB - We show how non-linear representations of local image structure can be used to improve the performance of model matching algorithms in medical image analysis tasks. Rather than represent the image structure using intensity values or gradients, we use measures that indicate the reliability of a set of local image feature detector outputs. These features are image edges, corners, and gradients. Feature detector outputs in flat, noisy regions tend to be ignored whereas those near strong structure are favoured. We demonstrate that combinations of these features give more accurate and reliable matching between models and new images than modelling image intensity alone. We also show that the approach is robust to non-linear changes in contrast, such as those found in multi-modal imaging. PMID- 15344464 TI - Knowledge-driven automated extraction of the human cerebral ventricular system from MR images. AB - This work presents an efficient and automated method to extract the human cerebral ventricular system from MRI driven by anatomic knowledge. The ventricular system is divided into six three-dimensional regions; six ROIs are defined based on the anatomy and literature studies regarding variability of the cerebral ventricular system. The distribution histogram of radiological properties is calculated in each ROI, and the intensity thresholds for extracting each region are automatically determined. Intensity inhomogeneities are accounted for by adjusting intensity threshold to match local situation. The extracting method is based on region-growing and anatomical knowledge, and is designed to include all ventricular parts, even if they appear unconnected on the image. The ventricle extraction method was implemented on the Window platform using C++, and was validated qualitatively on 30 MRI studies with variable parameters. PMID- 15344465 TI - Volumetric texture description and discriminant feature selection for MRI. AB - This paper considers the problem of classification of Magnetic Resonance Images using 2D and 3D texture measures. Joint statistics such as co-occurrence matrices are common for analysing texture in 2D since they are simple and effective to implement. However, the computational complexity can be prohibitive especially in 3D. In this work, we develop a texture classification strategy by a sub-band filtering technique that can be extended to 3D. We further propose a feature selection technique based on the Bhattacharyya distance measure that reduces the number of features required for the classification by selecting a set of discriminant features conditioned on a set training texture samples. We describe and illustrate the methodology by quantitatively analysing a series of images: 2D synthetic phantom, 2D natural textures, and MRI of human knees. PMID- 15344466 TI - CAD tool for burn diagnosis. AB - In this paper a new system for burn diagnosis is proposed. The aim of the system is to separate burn wounds from healthy skin, and the different types of burns (burn depths) from each other, identifying each one. The system is based on the colour and texture information, as these are the characteristics observed by physicians in order to give a diagnosis. We use a perceptually uniform colour space (L*u*v*), since Euclidean distances calculated in this space correspond to perceptually colour differences. After the burn is segmented, some colour and texture descriptors are calculated and they are the inputs to a Fuzzy-ARTMAP neural network. The neural network classifies them into three types of bums: superficial dermal, deep dermal and full thickness. Clinical effectiveness of the method was demonstrated on 62 clinical burn wound images obtained from digital colour photographs, yielding an average classification success rate of 82% compared to expert classified images. PMID- 15344467 TI - An inverse method for the recovery of tissue parameters from colour images. AB - The interpretation of colour images is presented as an inverse problem in which a mapping is sought between image colour vectors and the physiological parameters characterizing a tissue. To ensure the necessary one-to-one correspondence between the image colours and the parameters, the mapping must be unique. This can be established through testing the sign of the determinant of the Jacobian matrix, a multi-dimensional equivalent of a discrete derivative, over the space of all parameter values. Furthermore, an optimisation procedure is employed to find the set of filters for image capture which generate image vectors minimizing the mapping error. This methodology applied to interpretation of skin images shows that the standard RGB system of filters provides for a unique mapping between image values and parameters characterizing the normal skin. It is further shown that an optimal set of filters reduces the error of quantification by a factor of 2, on average. PMID- 15344468 TI - Ideal observer model for detection of blood perfusion and flow using ultrasound. AB - An ideal observer model is developed for the task of detecting blood perfusing or flowing through tissue. The ideal observer theory relies on a linear systems model that describes tissue and blood object functions and electronic noise as random processes. When aliasing is minimal, the system is characterized by a quantity similar to Noise-Equivalent Quanta used in photon imaging modalities. A simple 1-D model is used to illustrate the effect of the system and object parameters on task performance. Velocity and decorrelation are seen to be advantageous for detection. Aliasing can degrade performance. The ideal observer model provides a framework for assessing the performance of Power Doppler ultrasound systems, and may aid in their design. PMID- 15344469 TI - Permutation tests for classification: towards statistical significance in image based studies. AB - Estimating statistical significance of detected differences between two groups of medical scans is a challenging problem due to the high dimensionality of the data and the relatively small number of training examples. In this paper, we demonstrate a non-parametric technique for estimation of statistical significance in the context of discriminative analysis (i.e., training a classifier function to label new examples into one of two groups). Our approach adopts permutation tests, first developed in classical statistics for hypothesis testing, to estimate how likely we are to obtain the observed classification performance, as measured by testing on a hold-out set or cross-validation, by chance. We demonstrate the method on examples of both structural and functional neuroimaging studies. PMID- 15344470 TI - Ideal-observer performance under signal and background uncertainty. AB - We use the performance of the Bayesian ideal observer as a figure of merit for hardware optimization because this observer makes optimal use of signal-detection information. Due to the high dimensionality of certain integrals that need to be evaluated, it is difficult to compute the ideal observer test statistic, the likelihood ratio, when background variability is taken into account. Methods have been developed in our laboratory for performing this computation for fixed signals in random backgrounds. In this work, we extend these computational methods to compute the likelihood ratio in the case where both the backgrounds and the signals are random with known statistical properties. We are able to write the likelihood ratio as an integral over possible backgrounds and signals, and we have developed Markov-chain Monte Carlo (MCMC) techniques to estimate these high-dimensional integrals. We can use these results to quantify the degradation of the ideal-observer performance when signal uncertainties are present in addition to the randomness of the backgrounds. For background uncertainty, we use lumpy backgrounds. We present the performance of the ideal observer under various signal-uncertainty paradigms with different parameters of simulated parallel-hole collimator imaging systems. We are interested in any change in the rankings between different imaging systems under signal and background uncertainty compared to the background-uncertainty case. We also compare psychophysical studies to the performance of the ideal observer. PMID- 15344471 TI - Theoretical evaluation of the detectability of random lesions in Bayesian emission reconstruction. AB - Detecting cancerous lesion is an important task in positron emission tomography (PET). Bayesian methods based on the maximum a posteriori principle (also called penalized maximum likelihood methods) have been developed to deal with the low signal to noise ratio in the emission data. Similar to the filter cut-off frequency in the filtered backprojection method, the prior parameters in Bayesian reconstruction control the resolution and noise trade-off and hence affect detectability of lesions in reconstructed images. Bayesian reconstructions are difficult to analyze because the resolution and noise properties are nonlinear and object-dependent. Most research has been based on Monte Carlo simulations, which are very time consuming. Building on the recent progress on the theoretical analysis of image properties of statistical reconstructions and the development of numerical observers, here we develop a theoretical approach for fast computation of lesion detectability in Bayesian reconstruction. The results can be used to choose the optimum hyperparameter for the maximum lesion detectability. New in this work is the use of theoretical expressions that explicitly model the statistical variation of the lesion and background without assuming that the object variation is (locally) stationary. The theoretical results are validated using Monte Carlo simulations. The comparisons show good agreement between the theoretical predications and the Monte Carlo results. PMID- 15344472 TI - A unified statistical and information theoretic framework for multi-modal image registration. AB - We formulate and interpret several registration methods in the context of a unified statistical and information theoretic framework. A unified interpretation clarifies the implicit assumptions of each method yielding a better understanding of their relative strengths and weaknesses. Additionally, we discuss a generative statistical model from which we derive a novel analysis tool, the auto information function, as a means of assessing and exploiting the common spatial dependencies inherent in multi-modal imagery. We analytically derive useful properties of the auto-information as well as verify them empirically on multi modal imagery. Among the useful aspects of the auto-information function is that it can be computed from imaging modalities independently and it allows one to decompose the search space of registration problems. PMID- 15344473 TI - Information theoretic similarity measures in non-rigid registration. AB - Mutual Information (MI) and Normalised Mutual Information (NMI) have enjoyed success as image similarity measures in medical image registration. More recently, they have been used for non-rigid registration, most often evaluated empirically as functions of changing registration parameter. In this paper we present expressions derived from intensity histogram representations of these measures, for their change in response to a local perturbation of a deformation field linking two images. These expressions give some insight into the operation of NMI in registration and are implemented as driving forces within a fluid registration framework. The performance of the measures is tested on publicly available simulated multi-spectral MR brain images. PMID- 15344474 TI - A new & robust information theoretic measure and its application to image alignment. AB - In this paper we develop a novel measure of information in a random variable based on its cumulative distribution that we dub cumulative residual entropy (CRE). This measure parallels the well known Shannon entropy but has the following advantages: (1) it is more general than the Shannon Entropy as its definition is valid in the discrete and continuous domains, (2) it possess more general mathematical properties and (3) it can be easily computed from sample data and these computations asymptotically converge to the true values. Based on CRE, we define the cross-CRE (CCRE) between two random variables, and apply it to solve the image alignment problem for parameterized (3D rigid and affine) transformations. The key strengths of the CCRE over using the mutual information (based on Shannon's entropy) are that the former has significantly larger tolerance to noise and a much larger convergence range over the field of parameterized transformations. We demonstrate these strengths via experiments on synthesized and real image data. PMID- 15344475 TI - Gray scale registration of mammograms using a model of image acquisition. AB - A parametric technique is proposed to match the pixel-value distributions of two mammograms of the same woman. It can be applied to mammograms of the left and the right breast, or, more effectively, to temporal mammograms, e.g., from two screening rounds. The main reason to match mammograms is to lessen irrelevant differences between images due to acquisition: by varying breast compression, different film types, et cetera. Firstly, a technique like this might reduce the radiologist's efforts to detect relevant differences like abnormal growth in breast tissue that signals breast cancer. Secondly, though not the aim of this study, applications might be found in subtraction radiology or in the computer aided detection of abnormalities in temporal mammograms. Instead of arbitrarily shifting and/or scaling the pixel-values of one image to match the other or directly mapping one histogram to the other, the proposed method is based on general aspects of acquisition. This encompasses (1) breast compression; (2) exposure time; (3) incident radiation intensity; and, (4a) film properties and digitization for screen-film mammograms, or (4b) detector response for unprocessed digital mammograms. The method does not require a priori knowledge about specific settings of acquisition to match histograms; the degrees of freedom are estimated from the pixel-value distributions of the two mammograms themselves. By the method it is possible to match digitized screen-film mammograms (in the next also referred to as analog mammograms) as well as unprocessed digital mammograms in any of the four possible combinations: analog to analog, analog to digital, digital to analog, and digital to digital. PMID- 15344476 TI - Constructing diffeomorphic representations of non-rigid registrations of medical images. AB - The analysis of deformation fields, such as those generated by non-rigid registration algorithms, is central to the quantification of normal and abnormal variation of structures in the registered images. The correct choice of representation is an integral part of this analysis. This paper presents methods for constructing a general class of multi-dimensional diffeomorphic representations of deformations. We demonstrate that these representations are suitable for the description of deformations of medical images in 2 and 3 dimensions. Furthermore, we show that the non-Euclidean metric inherent in this representation is superior to the usual ad hoc Euclidean metrics in that it enables more accurate classification of legal and illegal variations. PMID- 15344477 TI - Topology preservation and regularity in estimated deformation fields. AB - A general formalism to impose topology preserving regularity on a given irregular deformation field is presented. The topology preservation conditions are derived with regard to the discrete approximations to the deformation field Jacobian in a two-dimensional image registration problem. The problem of enforcing topology preservation onto a given deformation field is formulated in terms of the deformation gradients, and solved using a cyclic projections approach. The generalization of the developed algorithm leads to a deformation field regularity control by limiting the per voxel volumetric change within a prescribed interval. Extension of the topology preservation conditions onto a three-dimensional registration problem is also presented, together with a comparative analysis of the proposed algorithm with respect to a Gaussian regularizer that enforces the same topology preservation conditions. PMID- 15344478 TI - Large deformation inverse consistent elastic image registration. AB - This paper presents a new image registration algorithm that accommodates locally large nonlinear deformations. The algorithm concurrently estimates the forward and reverse transformations between a pair of images while minimizing the inverse consistency error between the transformations. It assumes that the two images to be registered contain topologically similar objects and were collected using the same imaging modality. The large deformation transformation from one image to the other is accommodated by concatenating a sequence of small deformation transformations. Each incremental transformation is regularized using a linear elastic continuum mechanical model. Results of ten 2D and twelve 3D MR image registration experiments are presented that tested the algorithm's performance on real brain shapes. For these experiments, the inverse consistency error was reduced on average by 50 times in 2D and 30 times in 3D compared to the viscous fluid registration algorithm. PMID- 15344479 TI - Gaussian distributions on Lie groups and their application to statistical shape analysis. AB - The Gaussian distribution is the basis for many methods used in the statistical analysis of shape. One such method is principal component analysis, which has proven to be a powerful technique for describing the geometric variability of a population of objects. The Gaussian framework is well understood when the data being studied are elements of a Euclidean vector space. This is the case for geometric objects that are described by landmarks or dense collections of boundary points. We have been using medial representations, or m-reps, for modelling the geometry of anatomical objects. The medial parameters are not elements of a Euclidean space, and thus standard PCA is not applicable. In our previous work we have shown that the m-rep model parameters are instead elements of a Lie group. In this paper we develop the notion of a Gaussian distribution on this Lie group. We then derive the maximum likelihood estimates of the mean and the covariance of this distribution. Analogous to principal component analysis of covariance in Euclidean spaces, we define principal geodesic analysis on Lie groups for the study of anatomical variability in medially-defined objects. Results of applying this framework on a population of hippocampi in a schizophrenia study are presented. PMID- 15344480 TI - Non-rigid image registration using a statistical spline deformation model. AB - We propose a statistical spline deformation model (SSDM) as a method to solve non rigid image registration. Within this model, the deformation is expressed using a statistically trained B-spline deformation mesh. The model is trained by principal component analysis of a training set. This approach allows to reduce the number of degrees of freedom needed for non-rigid registration by only retaining the most significant modes of variation observed in the training set. User-defined transformation components, like affine modes, are merged with the principal components into a unified framework. Optimization proceeds along the transformation components rather then along the individual spline coefficients. The concept of SSDM's is applied to the temporal registration of thorax CR-images using pattern intensity as the registration measure. Our results show that, using 30 training pairs, a reduction of 33% is possible in the number of degrees of freedom without deterioration of the result. The same accuracy as without SSDM's is still achieved after a reduction up to 66% of the degrees of freedom. PMID- 15344481 TI - A view-based approach to registration: theory and application to vascular image registration. AB - This paper presents an approach to registration centered on the notion of a view- a combination of an image resolution, a transformation model, an image region over which the model currently applies, and a set of image primitives from this region. The registration process is divided into three stages: initialization, automatic view generation, and estimation. For a given initial estimate, the latter two alternate until convergence; several initial estimates may be explored. The estimation process uses a novel generalization of the Iterative Closest Point (ICP) technique that simultaneously considers multiple correspondences for each point. View-based registration is applied successfully to alignment of vascular and neuronal images in 2-d and 3-d using similarity, affine, and quadratic transformations. PMID- 15344482 TI - Fusion of autoradiographies with an MR volume using 2-D and 3-D linear transformations. AB - The recent development of 3-D medical imaging devices has given access to the 3-D imaging of in vivo tissues, from an anatomical (MR, CT) or even functional point of view (fMRI, PET, SPECT). However, the resolution of these images is still not sufficient to image anatomical or functional details, that can only be revealed by in vitro imaging (e.g. histology, autoradiography). The deep motivation of this work is the comparison of activations detected by fMRI series analysis to the ones that can be observed in autoradiographic images. The aim of the presented work is to fuse the autoradiographic data with the pre-mortem anatomical MR image, to facilitate the above-mentioned comparison. We show that this fusion can be achieved by using only simple global transformations (rigid and affine), yielding a very satisfactory result. PMID- 15344483 TI - Bayesian multimodality non-rigid image registration via conditional density estimation. AB - We present a Bayesian multimodality non-rigid image registration method. Since the likelihood is unknown in the general multimodality setting, we use a density estimator as a drop in replacement to the true likelihood. The prior is a standard small deformation penalty on the displacement field. Since mutual information-based methods are in widespread use for multimodality registration, we attempt to relate the Bayesian approach to mutual information-based approaches. To this end, we derive a new criterion which when satisfied, guarantees that the displacement field which minimizes the Bayesian maximum a posteriori (MAP) objective also maximizes the true mutual information (with a small deformation penalty) as the number of pixels tends to infinity. The criterion imposes an upper bound on the number of configurations of the displacement field. Finally, we compare the results of the Bayesian approach with mutual information, joint entropy and joint probability approaches on synthetic data and simulated T1 and T2 2D MR images. PMID- 15344484 TI - Spatiotemporal localization of significant activation in MEG using permutation tests. AB - We describe the use of non-parametric permutation tests to detect activation in cortically-constrained maps of current density computed from MEG data. The methods are applicable to any inverse imaging method that maps event-related MEG to a coregistered cortical surface. To determine an appropriate threshold to apply to statistics computed from these maps, it is important to control for the multiple testing problem associated with testing 10's of thousands of hypotheses (one per surface element). By randomly permuting pre- and post-stimuius data from the collection of individual epochs in an event related study, we develop thresholds that control the familywise (type 1) error rate. These thresholds are based on the distribution of the maximum intensity, which implicitly accounts for spatial and temporal correlation in the cortical maps. We demonstrate the method in application to simulated data and experimental data from a somatosensory evoked response study. PMID- 15344485 TI - Symmetric BEM formulation for the M/EEG forward problem. AB - The forward M/EEG problem consists in simulating the electric potential and the magnetic field produced outside the head by currents in the brain related to neural activity. All previously proposed solutions using the Boundary Element Method (BEM) were based on a double-layer integral formulation. We have developed an alternative symmetric BEM formulation, achieving a significantly higher accuracy for sources close to tissue interfaces, namely in the cortex. Numerical experiments using a spherical semi-realistic multilayer head model with a known analytical solution are presented, showing that the new BEM performs better than the formulations used in our earlier comparisons, and in most cases outperforms the Finite Element Method (FEM) as far as accuracy is concerned, thus making the BEM a viable choice. PMID- 15344486 TI - Localization Estimation Algorithm (LEA): a supervised prior-based approach for solving the EEG/MEG inverse problem. AB - Localizing and quantifying the sources of ElectroEncephaloGraphy (EEG) and MagnetoEncephaloGraphy (MEG) measurements is an ill-posed inverse problem, whose solution requires a spatial regularization involving both anatomical and functional priors. The distributed source model enables the introduction of such constraints. However, the resulting solution is unstable since the equation system one has to solve is badly conditioned and under-determined. We propose an original approach for solving the inverse problem, that allows to deal with a better-determined system and to temper the influence of priors according to their consistency with the measured EEG/MEG data. This Localization Estimation Algorithm (LEA) estimates the amplitude of a selected subset of sources, which are localized based on a prior distribution of activation probability. LEA is evaluated through numerical simulations and compared to a classical Weighted Minimum Norm estimation. PMID- 15344487 TI - Multivariate group effect analysis in functional Magnetic Resonance Imaging. AB - In functional MRI (fMRI), analysis of multisubject data typically involves spatially normalizing (i.e. co-registering in a common standard space) all data sets and summarizing results in a single group activation map. This widely used approach does not explicitely account for between-subject anatomo-functional variability. Therefore, we propose a group effect analysis method which makes use of a multivariate model to select the main signal variations that are common to all subjects, while allowing final statistical inference on the individual scale. The normalization step is thus avoided and individual anatomo-functional features are preserved. The approach is evaluated by using simulated data and it is shown that sensitivity is drastically improved compared to more conventional individual analysis. PMID- 15344488 TI - Meshfree representation and computation: applications to cardiac motion analysis. AB - For medical image analysis issues where the domain mappings between images involve large geometrical shape changes, such as the cases of nonrigid motion recovery and inter-object image registration, the finite element methods exhibit considerable loss of accuracy when the elements in the mesh become extremely skewed or compressed. Therefore, algorithmically difficult and computationally expensive remeshing procedures must be performed in order to alleviate the problem. We present a general representation and computation framework which is purely based on the sampling nodal points and does not require the construction of mesh structure of the analysis domain. This meshfree strategy can more naturally handle very large object deformation and domain discontinuity problems. Because of its intrinsic h-p adaptivity, the meshfree framework can achieve desired numerical accuracy through adaptive node and polynomial shape function refinement with minimum extra computational expense. We focus on one of the more robust meshfree efforts, the element free Galerkin method, through the moving least square approximation and the Galerkin weak form formulation, and demonstrate its relevancy to medical image analysis problems. Specifically, we show the results of applying this strategy to physically motivated multiframe motion analysis, using synthetic data for accuracy assessment and for comparison to finite element results, and using canine magnetic resonance tagging and phase contrast images for cardiac kinematics recovery. PMID- 15344489 TI - Visualization of myocardial motion using MICSR trinary checkerboard display. AB - Magnetic resonance tagging is used to quantify and visualize myocardial motion. The lack of tag persistence, however, is often a problem in visualizing motion throughout the cardiac cycle, particularly in late diastole. Complementary spatial modulation of magnetization (CSPAMM) improves tag persistence, but requires complex signal subtraction, which is not available on all scanners. This paper analyzes the contrast to noise properties of a new method called magnitude image CSPAMM reconstruction (MICSR), which requires only magnitude images from the scanner and demonstrates improved contrast and tag persistence over CSPAMM. A novel "trinary checkerboard" display of myocardial motion, ideally matched to the contrast to noise properties of MICSR, is also presented. PMID- 15344490 TI - Velocity estimation in ultrasound images: a block matching approach. AB - In this paper, we focus on velocity estimation in ultrasound images sequences. Ultrasound images present many difficulties in image processing because of the typically high level of noise found in them. Recently, Cohen and Dinstein have derived a new similarity measure, according to a simplified image formation model of ultrasound images, optimal in the maximum likelihood sense. This similarity measure is better for ultrasound images than others such as the sum-of-square differences or normalised cross-correlation because it takes into account the fact that the noise in an ultrasound image is multiplicative Rayleigh noise, and that displayed ultrasound images are log-compressed. In this work we investigate the use of this similarity measure in a block matching method. The underlying framework of the method is Singh's algorithm. New improvements are made both on the similarity measure and the Singh algorithm to provide better velocity estimates. A global optimisation scheme for algorithm parameter estimation is also proposed. We show that this optimisation makes an improvement of approximately 35% in comparison to the result obtained with the worst parameter set. Results on clinically acquired cardiac and breast ultrasound sequences, demonstrate the robustness of the method. PMID- 15344491 TI - Construction of a statistical model for cardiac motion analysis using nonrigid image registration. AB - In this paper we present a new technique for tracking the movement of the myocardium using a statistical model derived from the motion fields in the hearts of several healthy volunteers. To build the statistical model we tracked the motion of the myocardium in 17 volunteers using a nonrigid registration technique based on free-form deformations and mapped the motion fields obtained into a common reference coordinate system. A principal component analysis (PCA) was then performed on the motion fields to extract the major modes of variation in the fields between the successive time frames. The modes of variation obtained were then used to parametrize the free-form deformations and build our statistical model. The results of using our model to track the motion of the heart in normal volunteers are also presented. PMID- 15344492 TI - Fast tracking of cardiac motion using 3D-HARP. AB - MR tagging is considered as a valuable technique to evaluate regional myocardial function quantitatively and noninvasively, however the cumbersome and time consuming post-processing procedures for cardiac motion tracking still hinder its application to routine clinical examination. We present a fast and semiautomatic method for tracking 3D cardiac motion from short-axis (SA) and long-axis (LA) tagged MRI images. The technique, called 3D-HARP (HARmonic Phase), is based on the HARP method and extends this method to track 3D motion. A material mesh model is built to represent a collection of material points inside the left ventricle (LV) wall. The phase time-invariance property of material points is used to track the mesh points. For a series of 9 timeframe MRI images, the total time required for initializing settings, building the mesh, and tracking 3D cardiac motion is approximately 10 minutes. Further analysis of Langrangian strain and twist angle demonstrates that during systole, the lateral LV wall shows a greater strain values than the septum and the SA slices from the base to the apex show a gradual change in twist pattern. PMID- 15344493 TI - Analysis of event-related fMRI data using best clustering bases. AB - We explore a new paradigm for the analysis of event-related functional magnetic resonance images (fMRI) of brain activity. We regard the fMRI data as a very large set of time series x(i)(t), indexed by the position i of a voxel inside the brain. The decision that a voxel i0 is activated is based not solely on the value of the fMRI signal at i0, but rather on the comparison of all time series x(i)(t) in a small neighborhood Wi0 around i0. We construct basis functions on which the projection of the fMRI data reveals the organization of the time-series x(i)(t) into "activated", and "non-activated" clusters. These "clustering basis functions" are selected from large libraries of wavelet packets according to their ability to separate the fMRI time-series into the activated cluster and a non activated cluster. This principle exploits the intrinsic spatial correlation that is present in the data. PMID- 15344494 TI - Estimation of the Hemodynamic Response Function in event-related functional MRI: directed acyclic graphs for a general Bayesian inference framework. AB - A convenient way to analyze BOLD fMRI data consists of modeling the whole brain as a stationary, linear system characterized by its transfer function: the Hemodynamic Response Function (HRF). HRF estimation, though of the greatest interest, is still under investigation, for the problem is ill-conditioned. In this paper, we recall the most general Bayesian model for HRF estimation and show how it can beneficially be translated in terms of graphical models, leading to (i) a clear and efficient representation of all structural and functional relationships entailed by the model, and (ii) a straightforward numerical scheme to approximate the joint posterior distribution, allowing for estimation of the HRF, as well as all other model parameters. We finally apply this novel technique on both simulations and real data. PMID- 15344495 TI - Nonlinear estimation and modeling of fMRI data using spatio-temporal support vector regression. AB - This paper presents a new and general nonlinear framework for fMRI data analysis based on statistical learning methodology: support vector machines. Unlike most current methods which assume a linear model for simplicity, the estimation and analysis of fMRI signal within the proposed framework is nonlinear, which matches recent findings on the dynamics underlying neural activity and hemodynamic physiology. The approach utilizes spatio-temporal support vector regression (SVR), within which the intrinsic spatio-temporal autocorrelations in fMRI data are reflected. The novel formulation of the problem allows merging model-driven with data-driven methods, and therefore unifies these two currently separate modes of fMRI analysis. In addition, multiresolution signal analysis is achieved and developed. Other advantages of the approach are: avoidance of interpolation after motion estimation, embedded removal of low-frequency noise components, and easy incorporation of multi-run, multi-subject, and multi-task studies into the framework. PMID- 15344496 TI - A constrained variational principle for direct estimation and smoothing of the diffusion tensor field from DWI. AB - In this paper, we present a novel constrained variational principle for simultaneous smoothing and estimation of the diffusion tensor field from diffusion weighted imaging (DWI). The constrained variational principle involves the minimization of a regularization term in an LP norm, subject to a nonlinear inequality constraint on the data. The data term we employ is the original Stejskal-Tanner equation instead of the linearized version usually employed in literature. The original nonlinear form leads to a more accurate (when compared to the linearized form) estimated tensor field. The inequality constraint requires that the nonlinear least squares data term be bounded from above by a possibly known tolerance factor. Finally, in order to accommodate the positive definite constraint on the diffusion tensor, it is expressed in terms of cholesky factors and estimated. variational principle is solved using the augmented Lagrangian technique in conjunction with the limited memory quasi-Newton method. Both synthetic and real data experiments are shown to depict the performance of the tensor field estimation algorithm. Fiber tracts in a rat brain are then mapped using a particle system based visualization technique. PMID- 15344497 TI - Persistent Angular Structure: new insights from diffusion MRI data. Dummy version. AB - We determine a statistic called the radially Persistent Angular Structure (PAS) from samples of the Fourier transform of a three-dimensional function. The method has applications in diffusion magnetic resonance imaging (MRI), which samples the Fourier transform of the probability density function of particle displacements. The persistent angular structure is then a representation of the relative mobility of particles in each direction. In combination, PAS-MRI computes the persistent angular structure at each voxel of an image. This technique has biomedical applications, where it reveals the orientations of microstructural fibres, such as white-matter fibres in the brain. We test PAS-MRI on synthetic and human brain data. The data come from a standard acquisition scheme for diffusion-tensor MRI in which the samples in each voxel lie on a sphere in Fourier space. PMID- 15344499 TI - My two cents worth. PMID- 15344498 TI - Probabilistic Monte Carlo based mapping of cerebral connections utilising whole brain crossing fibre information. AB - A methodology is presented for estimation of a probability density function of cerebral fibre orientations when one or two fibres are present in a voxel. All data are acquired on a clinical MR scanner, using widely available acquisition techniques. The method models measurements of water diffusion in a single fibre by a Gaussian density function and in multiple fibres by a mixture of Gaussian densities. The effects of noise on complex MR diffusion weighted data are explicitly simluated and parameterised. This information is used for standard and Monte Carlo streamline methods. Deterministic and probabilistic maps of anatomical voxel scale connectivity between brain regions are generated. PMID- 15344500 TI - A brief history of clinical quality and medical errors. PMID- 15344501 TI - The physician's role in the public reporting of hospital quality data. PMID- 15344502 TI - Paying for quality. PMID- 15344503 TI - Building a system for improvement: the experience of McLeod Health. PMID- 15344504 TI - Training resident physicians toward improved patient safety. AB - Residents in training represent both a problem and opportunity in promoting enhanced patients safety in acute care settings. Trainees do not bring the experience and knowledge of attending physicians to the bedside and have an increased risk for medical errors. Yet, the around-the-clock presence of residents in the hospital and their awareness of impediments to safer patient care present a potential for preventing error-related adverse events. Capturing this potential for improved patient safety requires a broadened training curriculum that focuses on patient safety. PMID- 15344505 TI - Keeper of the bar. PMID- 15344506 TI - Quality's perverse economics. PMID- 15344507 TI - Clinical quality and patient safety. PMID- 15344509 TI - Fertility and the freezer. New technology allows women to put their eggs on ice. Doctors urge caution, but some people can't wait. PMID- 15344508 TI - The Village Quack. PMID- 15344510 TI - Physiological changes in largemouth bass exposed to paper mill effluents under laboratory and field conditions. AB - We report here on studies designed to asses the effects of paper mill effluents on non-reproductive functions of free-ranging and captive Florida largemouth bass (Micropterus salmoides floridanus). This was accomplished by conducting an outdoor tank study, in which fish were exposed to well water or to 10%, 20%, 40%, and 80% full strength effluent for 28 or 56 days, and by sampling largemouth bass from sites within the St. Johns River, Florida, upstream and downstream from a paper mill plant. Blood and plasma samples from fish from the tank study and from fish sampled from the ambient sites were analyzed for over 20 variables. We also determined liver and spleen weights and examined them histologically. The most significant finding from the tank study was an increase in the concentration of albumin and hepatosomatic index for bass exposed to > or = 20% effluents for 56 days. Spleenosomatic index and number of melanomacrophage centers were decreased in bass from effluent-dominated sites (Palatka and Rice Creek), whereas concentrations of calcium, phosphorous, glucose, and creatinine were elevated in fish from these sites, compared to fish from reference streams. Fish from Rice Creek also had fewer red blood cells, and male bass from Palatka had lower concentrations of cholesterol. Plasma concentrations of albumin and hepatic concentrations of glutathione were elevated in males from Palatka, and both females and males from Rice Creek had higher concentrations of globulin. These results indicate a complex pattern of effects of paper mill effluents on several physiological functions. However, despite the myriad of treatment and site related effects, most physiological parameters fell within normal ranges when compared to reports on largemouth bass and other freshwater species. PMID- 15344511 TI - Molecular characterization of contaminant-indicative RAPD markers. AB - This paper describes genetic markers which can be used to study selection and genetic adaptation of organisms to radionuclide and other types of contaminant stress. Previous research using the randomly amplified polymorphic DNA (RAPD) technique has identified several markers which revealed genetic differences between contaminated and reference western mosquitofish (Gambusia affinis) populations. Experimental evidence suggested that these markers may be associated with loci involved in determining relative fitness in radionuclide-contaminated environments ("contaminant-indicative markers"). In the present study, Southern blot analyses show these markers to be highly conserved in DNA sequence and molecular length in sea urchins, mosquitofish, herring gulls and humans. Such conservation is thought to be rare among RAPD bands. Results of DNA sequencing efforts did not provide definitive evidence as to the identity of these loci, but indicated that short segments (<40 bp) of known DNA sequences were homologous to various regions of the RAPD sequences. Furthermore, the regions of homology seemed to be non-randomly distributed along the length of the RAPD markers. Although the identity of these bands is still unknown, the high degree of conservatism suggests that these loci might play an important role in molecular processes. PMID- 15344512 TI - The impact of ballast water effluent on the Manila clam, Tapes semidecussatus. AB - The impact of ballast water discharge from an oil refinery in Whitegate Harbour, Co. Cork, Ireland to the Manila clam, Tapes semidecussatus, was investigated using a whole sediment bioassay. Bioassay organisms were exposed to surface sediments for a period of 21 days, including a reference and control sediment, collected from the vicinity of a ballast water discharge pipe. At the whole organism level, mortality and burrowing behaviour (assessed as time taken for organisms to burrow into the sediment) were used to determine the effects of exposure to test sediments. After 21 days, clam mortality was higher in surface sediments collected directly at the point of the ballast water discharge pipe than from all other test sediments assayed. After 10 days exposure to test sediments collected from the vicinity of the ballast water discharge pipe, the burrowing behaviour of animals was significantly different to the behaviour of animals exposed to uncontaminated sediments. PMID- 15344513 TI - Assessment of sediment ecotoxicity and genotoxicity in freshwater laboratory microcosms. AB - One possible fate of dredged sediments is disposal in a submerged gravel pit, but this practice may be a source of contamination of the aquatic environment. In an attempt to assess the risks for the aquatic ecosystem, a microcosm test was conducted in the laboratory on three sediments of channel sections to be dredged. After a 3-day period of stabilization, pelagic organisms (microalgae, daphnids, duckweeds, pond snails) and benthic organisms (amphipods, chironomids) were introduced into microcosms. Survival and growth of organisms were monitored during 28 days. Genetic damage was measured weekly in pond snail hemocytes through the comet assay. The sediments contained high levels of metals (up to 740 mg copper/kg and 1220 mg zinc/kg), but were moderately contaminated by PAHs (total PAH content < 10 mg/kg) and PCBs (total PCB content < 0.6 mg/kg). Only moderate effects on the biota were observed, except for genetic damage on pond snail hemocytes which was significant in all microcosms. The absence of pronounced toxic effects on organisms was not expected since concentrations of some sediment toxicants were above threshold values reported in the literature, especially for metals in two sediments, and for PAHs in the three sediments. Speciation, influence of organic matter and presence of sulfides might explain the low toxicity observed. By contrast, assessment of DNA damage revealed to be more sensitive and enabled a ranking of sediments coherent with their toxic load. In the present work, the microcosm protocol previously set up with a formulated sediment appears to be fitted to natural sediments. PMID- 15344514 TI - The responses of selected terrestrial plants to short (<12 days) and long term (2, 4 and 6 weeks) hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) exposure. Part I: Growth and developmental effects. AB - Soils contaminated with explosive materials like hexahydro-1,3,5-trinitro-1,3,5 triazine (RDX) is a concern nation-wide on military installations and sites where explosives are manufactured, stored, or disposed. Terrestrial plants are a vital group of receptor organism, yet limited published information is available on the potential impacts of RDX exposure in terrestrial plants. This research comprised the initial phases in the development of a short-term (<12 days) screening experiment for assessing the environmental impacts of RDX exposure in terrestrial plants. Fifteen plants (dicots and monocots) were exposed to three soils amended with 0-4000 microg g(-1) of RDX during the short-term screening experiments. Growth responses (maximum root and shoot lengths, percent emergence) and adverse developmental effects were the assessment endpoints. Sunflower was identified as the most RDX sensitive plant and selected for evaluation during the long-term (2, 4, and 6 weeks) experiments. Two life stages of sunflower (embryos and 2-week old seedlings) were exposed to Grenada soil amended with 0-100 microg g(-1) of RDX. The assessment endpoints during the long-term experiments included: biomass, maximum shoot and root length, root bio-volume, maximum stem diameter, number of leaves, and adverse developmental effects. Statistically significant differences were measured in several of the growth parameters following the short and long term exposure studies, however there were no consistent patterns. The consistent indicators of detrimental impacts from RDX exposure were the adverse developmental effects observed, regardless of life stage, soil type, or exposure duration. Typically, more adverse developmental effects were observed in dicots than monocots. The efficacy of the short-term screening experiments for estimating the impacts of long-term RDX exposure was validated. PMID- 15344516 TI - Orthopaedic care in spina bifida: past, present, and future. PMID- 15344515 TI - Toxicity testing of fifteen non-crop plant species with six herbicides in a greenhouse experiment: implications for risk assessment. AB - Estimation of risk to plants not targeted by herbicides when used in agricultural or forestry situations requires appropriate data on multiple species. Currently, many questions remain unresolved as to the adequate type and number of species to be tested. This paper presents the result of a unique greenhouse experiment where testing was performed with 15 non-crop plant species sprayed with 6 herbicides. The herbicides were chosen because of their different modes of action and because they are widely used in several countries. The plants favoured were species commonly found in field margins of Europe and/or North America. This dataset (called thereafter Danish/Canadian) was compared to the crop species that had been submitted to the US EPA for the same herbicides. In general, the selected plant species in the Danish/Canadian database were easy to grow and maintain in the greenhouse. The Danish/Canadian plants were overall more sensitive than the species tested in the US EPA data, yielding to a 5% protection threshold (HC5(50)) that was always more conservative. There was a large variability in plant responses among herbicides. Recommendations are provided on species that should and should not be used for risk assessment of non-target plants. PMID- 15344517 TI - Motor impairments in young children with cerebral palsy: relationship to gross motor function and everyday activities. AB - In this study we assessed the distribution of spasticity, range of motion (ROM) deficits, and selective motor control problems in children with cerebral palsy (CP), and examined how these impairments relate to each other and to gross motor function and everyday activities. Ninety-five children (55 males, 40 females; mean age 58 months, SD 18 months, range 25 to 87 months) were evaluated with the modified Ashworth scale (MAS), passive ROM, the Selective Motor Control scale (SMC), the Gross Motor Function Measure (GMFM), and the Pediatric Evaluation of Disability Inventory (PEDI). Types of CP were hemiplegia (n=19), spastic diplegia (n=40), ataxic diplegia (n=4), spastic quadriplegia (n=16), dyskinetic (n=9), and mixed type (n=7). Severity spanned all five levels of the Gross Motor Function Classification System (GMFCS). The findings highlight the importance of measuring spasticity and ROM in several muscles and across joints. Wide variability of correlations of MAS, ROM, and SMC indicates a complex relationship between spasticity, ROM, and selective motor control. Loss of selective control seemed to interfere with gross motor function more than the other impairments. Further analyses showed that motor impairments were only one component among many factors that could predict gross motor function and everyday activities. Accomplishment of these activities was best predicted by the child's ability to perform gross motor tasks. PMID- 15344518 TI - Comparison of three ankle-foot orthosis configurations for children with spastic diplegia. AB - This study compared the functional efficacy of three commonly prescribed ankle foot orthosis (AFO) configurations (solid [SAFO], hinged [HAFO], and posterior leaf spring [PLS]). Sixteen independently ambulatory children (10 males, six females; mean age 8 years 4 months, SD 2 years 4 months; range 4 years 4 months to 11 years 6 months) with spastic diplegia participated in this study. Four children were classified at level I of the Gross Motor Function Classification System (GMFCS; Palisano et al. 1997); the remaining 12 were at level II. Children were assessed barefoot (BF) at baseline (baseline assessment of energy consumption was performed with shoes on, no AFO) and in each orthotic configuration after three months of use, using gait analysis, oxygen consumption, and functional outcome measures. AFO use did not markedly alter joint kinematics or kinetics at the pelvis, hip, or knee. All AFO configurations normalized ankle kinematics in stance, increased step/stride length, decreased cadence, and decreased energy cost of walking. Functionally, all AFO configurations improved the execution of walking/running/jumping skills, upper extremity coordination, and fine motor speed/dexterity. However, the quality of gross motor skill performance and independence in mobility were unchanged. These results suggest that most children with spastic diplegia benefit functionally from AFO use. However, some children at GMFCS level II demonstrated a subtle but detrimental effect on function with HAFO use, shown by an increase in peak knee extensor moment in early stance, excessive ankle dorsiflexion, decreased walking velocity, and greater energy cost. Therefore, constraining ankle motion by using a PLS or SAFO should be considered for most, but not all, children with spastic diplegia. PMID- 15344519 TI - Non-verbal development of children with deafness with and without cochlear implants. AB - Deprivation of sensory input affects neurological development. Our objective was to explore clinically the role of hearing in development of sensorimotor integration and non-verbal cognition. The study involved 54 children (15 males, 839 females; 5 to 9 years old) with severe or profound bilateral prelocutive deafness but without neurological or cognitive impairment. Of these, 25 had received an early cochlear implant (CIm). Patients were compared with 40 children with normal hearing. All were given a battery of non-verbal neuropsychological tests and a balance test, and were timed for simple and complex movement of limbs. Deafness, whether treated by CIm or not, resulted in a delay in development of complex motor sequences and balance. Lack of auditory input was also associated with lower, but non-pathological, scores in visual gnoso-praxic tasks and sustained attention. Such differences were not observed in children with CIm. Hearing contributes to clinical development of spatial integration, motor control, and attention. An early CIm enables good verbal development and might also improve non-verbal capacities. PMID- 15344520 TI - A volumetric study of parietal lobe subregions in Turner syndrome. AB - Turner syndrome, a genetic disorder that results from the complete or partial absence of an X chromosome in females, has been associated with specific impairment in visuospatial cognition. Previous studies have demonstrated a relationship between parietal lobe abnormalities and visuospatial deficits in Turner syndrome. We used high-resolution magnetic resonance imaging to measure parietal lobe subdivisions in 14 participants with Turner syndrome (mean age 13 years 5 months, SD 5 years) and 14 age-matched controls (mean age 13 years 5 months, SD 4 years 7 months) to localize neuroanatomical variations more closely. Scans were acquired and analyzed for 14 females with Turner syndrome. Analyses of variance were used to investigate differences in regional parietal lobes. Females with Turner syndrome showed a bilateral parietal lobe reduction, specifically in the superior parietal and postcentral gyri. Full-scale IQ scores were significantly positively correlated with postcentral tissue volume in the Turner syndrome group. Structural differences in the parietal lobe are localized specifically to the anterior and superior parietal lobe and might be related to the visuospatial and visuomotor deficits associated with Turner syndrome. PMID- 15344521 TI - Sonographic appearance of the normal and abnormal insula of Reil. AB - Three planes are discerned during ultrasonographic screening of the insula in parasagittal view: opercular, insular, and fissural. Six newborn infants with normal brain anatomy, including two each of 28, 34, and 40 weeks' gestation, were selected for a description of the evolution of these parasagittal planes. Opercularization of the insula begins to be detected on a sonogram at about the 24th gestational week and progresses cranially. On coronal section the insular space forms a shallow groove at 24 weeks, becoming a slit at 28 weeks that grows longer and develops branches after 32 weeks. At 28 weeks the ascending anterior branch of the circular groove and the lateral fissure at the bottom of the insula are clearly seen in parasagittal section. Subsequent change consists of undulation and bifurcation of the lateral fissure, together with elongation of the anterior margin. Secondary gyri become visible in the insular dome between 28 and 34 weeks, forming short anterior and long posterior insular gyri. Five term newborn infants with perisylvian polymicrogyria were observed with both neonatal ultrasound and magnetic resonance imaging: two of unknown cause, and one each due to cytomegalovirus, bifunctional peroxisomal protein deficiency, and monozygous twinning. With polymicrogyria, on parasagittal sonographic examination at fissural level the anterior margin of the circular groove is a short rudiment. In insular view, only rudimentary sulci ascend from the lateral fissure; short and long gyri are not seen. In opercular view, no secondary branches are seen from the lateral fissure, an image akin to that seen at 28 weeks. A complete insular triangle is not recognized in any of these sections. A deep abnormal sulcus may prolong the lateral fissure when schizencephaly is associated with polymicrogyria. The insula in glutaric aciduria type II has no secondary gyri at term and is reduced to a simple and small triangle. Not all instances of polymicrogyria lead to macroscopically recognizable alteration of insular gyri. PMID- 15344522 TI - Congenital suprabulbar palsy: a distinct clinical syndrome of heterogeneous aetiology. AB - Congenital suprabulbar palsy is clinically characterized by problems of feeding, swallowing, drooling, and dysarthria. Epilepsy, delayed motor, cognitive, and language development, as well as learning disabilities may co-exist. Aetiology of the syndrome is diverse, yet studies often attribute it to specific entities. We report on nine patients (seven males, two females; age range 2 to 20 years), highlighting the heterogeneous causes of suprabulbar palsy using neuroimaging and emphasizing the need for systematic investigation for early detection and management. We identified patients with symmetrical infarcts involving the perisylvian region, apart from already-recognized conditions, such as congenital bilateral perisylvian syndrome (CBPS; a neuronal migration disorder) and Worster Drought syndrome. CBPS simulates Foix-Chavany-Marie syndrome in adults because of staged stroke but differs in many respects. Anoxia or ischemia to the developing brain could be a common plausible aetiology. Studies with large groups of patients are required to differentiate the various subgroups and identify essential criteria for diagnosis. PMID- 15344523 TI - Arteriovenous malformations presenting with papilloedema. AB - Cerebral arteriovenous malformations (AVMs) are fairly common and the majority of paediatric patients with this condition also present with intracranial haemorrhage. Two patients who had an incidental finding of an AVM associated with papilloedema are described here. The first was a 13-year-old male who presented after an accidental kick to the eyes. Examination revealed bilateral papilloedema. He gave a 2-year history of intermittent headache. Brain magnetic resonance imaging (MRI) showed an unruptured AVM in the temporal lobe. Lumbar puncture revealed elevated cerebrospinal fluid pressure. Visual acuity and visual fields were normal. He was treated with acetazolamide and improved within a few weeks. He subsequently underwent stereotactic radiosurgery to the AVM. He discontinued acetazolamide due to adverse side effects and there was no recurrence of headache and papilloedema. The second patient was a 14-year-old male who had polyarticular juvenile chronic arthritis and received low-dose steroids and methotrexate. Bilateral papilloedema was discovered during routine ophthalmology surveillance and he was otherwise asymptomatic neurologically. Brain MRI revealed an AVM in the posterior fossa. He had three embolization procedures, which have resulted in significant reduction in lesion size. The papilloedema resolved completely after the first two procedures, and visual acuity and fields remained normal. Here, possible underlying mechanism of raised intracranial pressure and importance of visual assessment in those with AVMs and their management are discussed. PMID- 15344524 TI - Effects of surgical adductor releases for hip subluxation in cerebral palsy: an AACPDM evidence report. PMID- 15344525 TI - From genetous ailments to genetic disorders: Ireland's On Idiocy and Imbecility Revisited. PMID- 15344526 TI - Quality of life of primary caregivers of children with cerebral palsy: a controlled study with Short Form-36 questionnaire. PMID- 15344527 TI - [GERD. Introduction]. AB - In Japan prevalence of GERD is still low as compared to Western countries. Although it is almost certain that the number of GERD patients will not increase by increasing patients who will receive eradication in Western countries, it may be the case in Japan, because most of the H. pylori-infected subjects in Japan have a considerable level of mucosal atrophy, that can be improved after eradication of H. pylori. Indeed, prevalence of H. pylori in Japanese patients with GERD is reported to be approximately 30-40%, suggesting protective role of H. pylori infection in the development of GERD. The disease entity of Non-erosive GERD (NERD) is receiving great attention recently, but effective rate of PPI for NERD is reported to be 60%, suggesting that a considerable number of non-GERD patients are included in patients diagnosed as having NERD. Thus, an effort for improving both sensitivity and specificity of the diagnosing method for NERD is required. In any event, it should be kept in mind that pathophysiology of GERD is quite different between Western and Japanese patients. PMID- 15344528 TI - [Reflux esophagitis--Japan and Western countries]. AB - Although reflux esophagitis (RE) is a common disease of the upper gastrointestinal tract in Western countries, its prevalence is still low in Asian countries, including Japan. On the other hand, the prevalence of H. pylori infection and atrophic gastritis are very high in Japan in comparison to that of other developed countries. Atrophic gastritis decreases gastric acid secretion, which is much lower in the Japanese compared to that in Western persons. The main mechanism of RE is reflux of gastric acid into the esophagus from the stomach. A decrease in gastric acid secretion induced by atrophic gastritis might prevent this acid reflux. This may be the reason RE is an uncommon disease in Japan. PMID- 15344529 TI - [GERD guideline]. AB - GERD guideline 2002 was defined by the experts in Japan after GERD Guideline Workshop 2002, based on the increased incidence of GERD in clinical practice in Japan and concentration of articles from Japanese patients. In the workshop, GERD was defined as follows; Gastroesophageal reflux disease (GERD) indicates those who have physical complications due to gastro-esophageal reflux and/or those who have impairment in their healthy life due to symptoms related to gastroesophageal reflux. In the diagnostic arm, evaluation of subjective symptom was placed as the most important. In the therapeutic arm, proton pump inhibitor (PPI) was named as the first line choice of medicine. PMID- 15344530 TI - [Pathophysiology of GERD: mechanisms of gastroesophageal reflux and prolonged esophageal acid exposure time]. AB - GERD is characterized by excessive esophageal acid exposure time. This suggests that either the rate of gastroesophageal reflux (GER) is higher and/or that the esophageal acid clearance time is longer. Transient LES relaxation (TLESR) is the single most common mechanism underlying GER in both normal subjects and patients with GERD. Whether or not the rate of TLESRs is higher in patients with GERD remains unclear. It is in the sitting or upright position that acid reflux mainly occurs, however, there seems to be no difference in the rate of TLESRs between both groups. The rate of TLESRs accompanied by acid reflux has been consistently shown to be significantly greater in patients with GERD than in normal subjects. Other mechanisms of reflux in patients with severe GERD are a hypotensive LES and ineffective esophageal motility which is found in severe GERD and which impairs bolus clearance of acid and thus increases acid contact time with the esophageal mucosa. PMID- 15344531 TI - [Reflux of duodenal or gastroduodenal contents induces esophageal carcinoma in rats]. AB - We observed the sequential development of columnar lined epithelium associated with adenocarcinoma, squamous dysplasia related with squamous cell carcinoma and adenosquamous carcinoma which were induced by duodeno-esophageal or gastro duodeno-esophageal reflux in rats. Wistar male rats, weighing approximately 250 g were employed. Animals received total gastrectomy and were reconstructed with esophago-jejunostomy, which causes unavoidable duodeno-esophageal reflux. The animals were sacrificed every 10 weeks after surgery until 50 weeks. Erosions and basal cell hyperplasia were observed in the lower esophageal squamous epithelium at 10 weeks after surgery. At 20 weeks, glandular structures featured with galactose oxidase-Schiff-positive staining (foveolar metaplasia) appeared in the basal layer of esophageal squamous epithelium. At 30 weeks, the glands developed and formed cysts which were stained with concanavalin A (pyloric glandular metaplasia) or/and high iron diamine and alcian blue (intestinal metaplasia). Since 40 weeks after surgery, esophageal carcinomas were found. As adenocarcinomas were surrounded by the columnar-lined epithelium, squamous cell carcinoma and adenosquamous carcinoma were accompanied by squamous dysplasia. Persistent duodeno-esophageal reflux can change the stem cells of squamous epithelium to make columnar-lined cells. As part of the sequence of events leading to the development of columnar-lined epithelium, foveolar metaplasia was observed followed by the appearance of pyloric glandular metaplasia and intestinal metaplasia. Chronic duodenal reflux induces the development of esophageal carcinoma not only adenocarcinoma also squamous cell carcinoma and adenosquamous carcinoma. These pathways of carcinogenesis were different dual patterns. PMID- 15344532 TI - [GERD and tight junction proteins of the esophageal mucosa]. AB - Recent advancement in the research of GERD has revealed that endoscopy negative GERD may not be a milder form of erosive GERD and may have different pathogenesis. We have previously proven that hypersensitivity to the acid of the esophageal mucosa plays an important role in its pathogenesis. Regarding the mechanisms for the esophageal hypersensitivity, we hypothesized that the tight junction proteins of the esophageal mucosa are fully or partially impaired in GERD patients. Accordingly, we immunohistologically studied the expression of various tight junction proteins using the rat reflux esophagitis model. The results demonstrated that the several kinds of tight junction proteins are expressed differently in the various parts of esophagus and their expression altered according to the development of reflux esophagitis. PMID- 15344533 TI - [Pro-inflammatory chemokine IL-8 in gastroesophageal reflux disease]. AB - Recent reports have demonstrated significantly higher expression levels of interleukin-8 (IL-8) in patients with gastroesophageal reflux disease (GERD) including non-erosive reflux disease (NERD). The levels of IL-8 mRNA expression were significantly decreased after proton pump inhibitor. The esophageal expression of CINCs, rat IL-8-like chemokines, was markedly enhanced in the models of acute or chronic esophagitis in rats. The production of IL-8 from esophageal mucosal cells was enhanced by the exposure to bile acid. These results suggest that IL-8 chemokine may play a major role in the pathogenesis of esophageal inflammation in GERD. PMID- 15344534 TI - [Diagnosis of gastro esophageal reflux disease and reflux esophagitis by using the questionnaire]. AB - QUEST can be carried out in a short time and having high susceptibility to the patients with reflux esophagitis. QUEST is also useful as a diagnostic tool for the patients with GERD. PMID- 15344535 TI - [Endoscopic findings of GERD]. AB - Reflux esophagitis is divided into four grades, i.e. Grade A to D, according to the severity of its mucosal break in the endoscopic classification of reflux esophagitis (Los Angels classification). In our study, only 14.3% of patients with heartburn had Grade A to D during endoscopy. This means that Los Angels classification is insufficient at least in Japan. Then we have claimed that Grade M and Grade N are to be included in this classification, which mean minimal change and no lesions, respectively. PMID- 15344536 TI - [Significance of PPI-test in the diagnosis of GERD]. AB - Gastroesophageal reflux disease (GERD) is defined as 'Chronic symptoms or mucosal damage produced by the abnormal reflux of gastric contents into the esophagus'. Reflux esophagitis refers to a subgroup of GERD patients with histopathologically demonstrated characteristic changes in the esophageal mucosa. Besides, GERD includes symptoms without endoscopic findings (endoscopic negative GERD) and extra-esophageal symptoms. Therefore, GERD cannot be diagnosed only by endoscopy. Three methods are indispensable in the diagnosis of GERD; endoscopy, evaluation of patient symptoms and acid reflux. Since 'Symptom relief is well correlated with the degree or suppression of gastric acid secretion in GERD', symptom in relation to acid reflux can be evaluated by PPI-test. Characteristics of PPI-test including extra-esophageal GERD diagnosis are discussed in this review. PMID- 15344537 TI - [Test of refractory gastroesophageal reflux disease]. AB - It should be considered that the causes of refractory gastroesophageal reflux disease (GERD) are multifactorial. Esophageal manometry study is useful when we make distinguish patients with esophageal motility disorders from those with refractory GERD. Endoscopic ultrasonography is also performed to observe the thickness of esophageal wall which represents the disturbance of esophageal motor function. Esophageal pH monitoring is useful to detect the acid clearance disturbance and phenomenon of nocturnal acid breakthrough. Both are occurred at night, and are recently considered to be responsible for refractory GERD. Catheter-free pH monitoring system, Bravo, makes it possible to measure esophageal pH under quite physiological conditions. Genotype of CYP2C19 is sometimes checked in patients with PPI resistance GERD. Intra-gastric pH with omeprazole and lansoprazole depends on patient's genotype of CYP2C19. Monitoring of 24-hour bilirubin, Bilitec, is also useful to detect duodeno-gastro-esophageal reflux. PMID- 15344538 TI - [Histopathological examination in gastroesophageal reflux disease (GERD)]. AB - We performed an immunohistochemical analysis of cell cycle-regulating protein (p21, p27 and Ki67) expression in endoscopic biopsy samples from the patients with gastroesophageal reflux disease (GERD) using angled-biopsy forceps. Inflammatory cell accumulation into the lamina propria was detected even in patients with modified Los Angeles (IA) system grades N and/or M. In grade N and M patients with no changes in the epithelium, the area of p21, p27 and Ki67 positive cells was expanded compared to normal mucosa. The area of p21, p27 and Ki67 positive cells tended to expand upward in the epithelium with GERD severity based on the LA classification grading. These indicate that inflammatory cell infiltration into the lamina propria is initial histological change of GERD. PMID- 15344539 TI - [Differential diagnosis of GERD]. AB - It was widely accepted that the prevalence of GERD is lower in Oriental countries compared to Western countries. But the incidence of GERD has recently increased in Japan. The most commonly recognized manifestation of GERD is heartburn or a substernal burning sensation in the chest. Most patients with reflux esophagitis complain of typical symptoms such as heartburn, regurgitation or dysphagia. However, some patients complain of atypical symptoms such as hoarse voice, chronic cough, adult-onset asthma or vocal cord polyps. It is not always easy to diagnose atypical symptomatic patients as GERD. If patients who complain of these atypical symptoms have not improved with common medical treatment, GERD should be the consideration in its differential diagnosis. PMID- 15344540 TI - [Primary care for GEDR]. AB - Current approach for the treatment of gastroesopageal reflux disease (GERD) was reviewed. The most effective treatment of erosive esophagitis or symptomatic GERD is to reduce gastric acid secretion with either an H2 receptor antagonists (H2RA) or a proton pump inhibitor (PPI). The PPI lead to more rapid healing and symptom relief than H2RA. Despite treatment with PPI, some patients with GERD continue to have symptoms or endoscopic evidence of esophagitis. Nocturnal acid breakthrough may be one of the mechanisms responsible for the refractory GERD. There are two approaches to the initial medical treatment of gastroesophageal reflux disease ('step down' therapy or 'step up' approach). Although there are arguments in favour of both approaches, the former is considered to be preferable these days. PMID- 15344541 TI - [Evaluation of the efficacy and the cost-effectiveness of maintenance treatment of gastroesophageal reflux disease: proton pump inhibitor versus histamine-2 receptor antagonist]. AB - GERD is a common condition and acid-suppressing agents are the mainstay of treatment. A cost-effectiveness analysis comparing a PPI, lansoprazole (LPZ) and a H2RA, famotidine (FAM) for the maintenance treatment of reflux esophagitis in Japan was performed using a Markov chain approach. The time period studied was 6 months and payer perspective was chosen. Transition probabilities were estimated from meta-analyses. Expected days without esophagitis (healthy days) were 166 for LPZ 30 mg/day, 161 for LPZ 15 mg/day and 143 for FAM 40 mg/day. Direct costs were 55,624 yen for LPZ 30 mg/day, 42,078 yen for LPZ 15 mg/day and 67,969 yen for FAM 40 mg/day. Cost-effectiveness ratio (direct costs/healthy days) was 335 yen for LPZ 30 mg/day, 262 yen for LPZ 15 mg/day and 477 yen for FAM 40 mg/day. Lansoprazole was superior to famotidine with regard to both efficacy and cost effectiveness and therefore is the preferred therapeutic agent for the maintenance treatment of GERD. PMID- 15344542 TI - [Pathogenesis and treatment of refractory gastroesophageal reflux disease in Japanese patients]. AB - Prevalence of refractory gastroesophageal reflux disease (GERD) defined as a patient who have persistent GERD symptoms during treatment with proton pump inhibitor (PPI) is rare in Japanese patinets. Pathogenesis of refractory GERD is associated with several factors including dysfunction of esophageal motility, presence of severe hiatal hernia, complication such as stricture and short esophagus, extensive metabolizer of CYP2C19 genotype, nocturnal gastric acid breakthrough, absence of H. pylori infection, or bile reflux. Examination by 24 hr pH monitoring is necessary to assess refractory GERD and if acid suppression is insufficient, treatment with double doses of PPIs or combination of PPI and H2 blocker is effective. However, most cases of refractory GERD are required surgical treatment. Endoscopic therapy might be useful for refractory GERD in future. PMID- 15344543 TI - [Surgical treatment for GERD]. AB - Surgical treatment for GERD is still indicated in a few cases. In particular, patients who resist PPI medical treatment, patients who do not comply well with medication, and young patients with strong subjective symptoms are candidates for surgical treatment. Others include patients with free reflux, with short esophagus, with esophageal stenosis, and with respiratory or oto-laryngological complications. The choice of operative method depends on the disease state and stage of GERD. The first choice of treatment in most cases is laparoscopic Nissen technique, and the second choice is laparoscopic Toupet method. The problem for surgeons is the need for extremely delicate technique in the reconstructive parts of the procedure in cases with a high degree of fundoplication and functional disorder. We adopted an operative technique that combines SPV, Toupet method and Hill method; we have found a good rate of patient acclimatization and high postoperative satisfaction. Time is still required, however, to bring laparoscopic surgery to the same level as open surgery. PMID- 15344544 TI - [Endoscopic treatment for gastroesophageal reflux disease]. AB - The endoscopic treatment as a minimally invasive method to gastroesophageal reflux disease (GERD) came to be tried in addition to medication and laparoscopic antireflux surgery. Clinical trials, such as the Endoluminal Gastroplication method, the Full Thickness Plicator method, the Stretta method, the Enteryx method, and the Gatekeeper method, are advancing in the United States and Europe. It is necessary to consider the efficacy, safety, durability, cost effectiveness, an indication, etc. using data from a randomized controlled trial with sufficient observation period. If the usefulness of endoscopic treatment is confirmed in the future, the new minimally invasive strategy over GERD will be established. PMID- 15344545 TI - [GERD: guidance in changing life-style]. AB - Gastroesophageal reflux disease (GERD) is increasing due to adoption of a western life style and an aging society in recent years. The symptoms of GERD, such as heartburn, regurgitation of gastric contents into the mouth, and chest pain, decrease QOL, and continuous treatment is necessary. Treatment of GERD is mainly drug therapy; however, it is believed that providing guidance in changing life style and providing information about the disease will permit long-lasting relief of the symptoms. PMID- 15344546 TI - [ENT manifestations of gastroesophageal reflux disease (GERD)]. AB - Gastroesophageal reflux disease (GERD) has been associated increasingly with ear, nose, and throat (ENT) signs and symptoms. However, the cause and effect relationship between these two clinical entities are far from established. Many patients diagnosed initially with GERD as the cause of laryngeal signs do not symptomatically or laryngoscopically respond to aggressive acid suppression and do not have abnormal esophageal acid exposure by pH monitoring. The hypothesis that GER causes laryngeal symptoms and conditions remains to be definitively proved. In many patients, the cause of laryngeal symptoms may well be multifactorial, and to identify definitively those patients in which GER may be playing a role remains a challenge. This has resulted in frustration on the part of both gastroenterologists and ENT physicians and confusion on the part of patients. In this article we discuss the recent data. PMID- 15344547 TI - [Chronic respiratory disease in patients with GERD]. AB - The association between gastroesophageal reflux disease and chronic respiratory diseases are proved in many report. Patient with GERD have a higher prevalence of asthma, chronic cough, lung fibrosis. Recently, it was reported patients with OSAS had significantly more GER event than controls. Antireflux therapy in asthma and chronic cough patient with GERD results in improvement in symptoms. In this paper, I comments and report about influence of GERD on chronic respiratory illness, and influence of an antireflux therapy for an asthmatic having GERD. PMID- 15344548 TI - [Differential diagnosis in GERD and cardiovascular disease]. AB - In substantial portion of patients referred to the coronary care unit (CCU) with a tentative diagnosis of acute coronary syndrome (ACS), gastro-esophageal diseases are the origins of their symptoms such as chest pain. The differential diagnosis is difficult and the coronary angiography (CAG) and the gastro esophageal endoscopy are often required. We recently evaluated our 100 consecutive cases that underwent CAG as ACS. Among the 100 cases, 72 had significant organic coronary artery lesions, 5 had significant coronary vasospasms proved by the acetylcholine provocation, and the other 23 showed no detectable major coronary artery abnormality. Among the last 23 cases, gastro esophageal endoscopy revealed the gastro-esophageal diseases such as reflux esophagitis in 7 cases. Those 7 cases with gastro-esophageal diseases had less coronary risk factors than the 77 cases with the coronary artery diseases. Gastro esophageal diseases are the major differential diagnoses of ACS, especially in those with few coronary risk factors. PMID- 15344549 TI - [Gastroesophageal reflux disease in diabetic patients]. AB - Esophageal dysfunctions occur frequently in patients with diabetic autonomic neuropathy, and the complication of gastroesophageal reflux disease (GERD) has also been reported. However, the characteristics of the GERD complicated with diabetes are obscure, because no detail assessment was performed. We recorded esophageal motility and acid reflux simultaneously in diabetic patients, and the correlation between esophageal dysfunction and diabetic neuropathy was examined. Esophageal dysfunctions including GERD were significantly related to diabetic motor neuropathy. Although the GERD is frequently complicated with diabetes, the symptoms are not apparent in diabetic patients. Therefore, physicians treating diabetic patients should have GERD in mind regardless of the symptoms. We also examined the effect of aldose reductase inhibitor (ARI) on the esophageal dysfunction in diabetic patients. Significant improvement of gastroesophageal reflux and esophageal motility were observed in diabetic patients by ARI treatment. ARI may be useful for the treatment of GERD complicated with diabetes. PMID- 15344550 TI - [Evaluation and treatment of gastroesophageal reflux in infants and children]. AB - Gastroesophageal reflux (GER), defined as passage of gastric contents into esophagus, and GER disease (GERD), defined as symptoms or complications of GER, are common pediatric problems encountered by both primary and specialty medical providers. Clinical manifestations of GERD in children include vomiting, poor weight gain, dysphagia, abdominal or substernal pain, esophagitis and respiratory disorders. On the other hand, recurrent vomiting is the symptom of hydronephrosis, brain tumor, food allergy, uremia, other metabolic disease, obstruction of intestine etc. It is very important for clinicians dealing with children and infants to understand GERD. The evaluation and treatment of gastroesophageal reflux in infants and children were reviewed here. PMID- 15344551 TI - [Recent view of gastro-esophageal reflux disease according to the guideline for gastric ulcer treatment]. AB - The guideline for gastric ulcer treatment in Japan recommends eradication of Helicobacter pylori (H. pylori) for the first choice. Recently, it is well known that some patients develop gastro-esophageal reflux disease (GERD) after successful eradication. H. pylori infection may play a protective role against GERD by impairing gastric acid secretion. Acid secretion is influenced by the distribution of gastritis in the stomach. Antrum-predominant gastritis is associated with gastric hypersecretion. Patients with corpus-predominant gastritis have decreased acid secretion. The latter is common in Japan and at high risk of GERD after eradication. To arise GERD, both acid secretion and reflux of acid caused by hiatus hernia or gastro-esophageal dysmotility are needed. Although most of GERD developed after eradication are mild, severe GERD are also experienced at times. Recent view of GERD and H. pylori infection is described. PMID- 15344552 TI - [Non-erosive GERD]. AB - Non-erosive reflux disease (NERD) is defined as those who have typical reflux symptoms without evidence of erosive changes in their lower esophageal mucosa. Symptoms should occur more than twice a week, and esophageal mucosa should be free from erosion. However, the differentiation from normal might be difficult because of the ambiguous definition. Pathophysiological mechanism on heartburn is not simply acidic stimuli to esophageal mucosa, but also esophageal distention, hypersensitive esophagus and/or exaggerate perception. NERD is not a simply milder form of reflux disease, because they have poor response to pharmacotherapy. Based on such evidence, newer concept for NERD is required. PMID- 15344553 TI - [PPI: new strategies for GERD]. AB - Proton pump inhibitor (PPI) is the first-line drug for GERD and is far more effective than H2-receptor antagonist (H2RA). H2RA suppresses mainly nocturnal gastric acid secretion from parietal cells, while PPI blocks acid production at nighttime as well as daytime when acid refluxes often occur. PPI-test is a therapeutic diagnosis and can reliably distinguish GERD from other diseases presenting similar symptoms. Initial therapy of GERD should be started with a full dose of PPI. However, most of the GERD patients need maintenance therapy. The maintenance dose of PPI should be individualized with a titration technique ('New Step-down therapy'). A small number of GERD patients resistant to PPI therapy may be due to nocturnal acid breakthrough (NAB) or rapid metabolism of PPI (extensive metabolizer for CYP2C19). PMID- 15344554 TI - [2,4-Dienoyl-CoA reductases: from discovery toward pathophysiological significance]. AB - 2,4-Dienoyl-CoA reductases from Escherichia coli and bovine liver were purified and characterized by the authors and almost simultaneously by Kunau and his colleagues around the early 1980s. The authors purified 2,4-dienoyl-CoA reductase from rat liver, cloned its cDNA and expressed its active form in Escherichia coli for the first time. Schulz and his colleagues have elaborately shown that mutant E. coli, in which the activity of the 2,4-dienoyl-CoA reductase is reduced to 12% of that in the parental strain, can grow on oleic acid but not on (Z)-6 octadecenoic acid as a sole carbon source. A single sporadic human fatality, in which the activities of 2,4-dienoyl-CoA reductases were reduced and accumulation of (2E,4Z) 2,4-decadienoylcarnitine was observed, has so far been reported. These taken together with the discovery of a novel delta3,5-delta2,4-dienoyl CoA isomerase and the absence of 3-hydroxyacyl-CoA epimerase have caused the classical metabolic pathway for beta-oxidation of unsaturated fatty acids, depicted by Stoffel in 1965, to be rewritten, and 2,4-dienoyl-CoA reductases are now known to be essential to beta-oxidation of unsaturated fatty acids. Very recently a single amino acid substitution adjacent to the NADPH binding site has been reported in pigs, suggesting that single nucleotide polymorphism will also be found to occur in humans. PMID- 15344555 TI - Th1/Th2 lymphocyte balance in patients with aplastic anemia. AB - Activated T cell plays an important role in the pathogenesis of aplastic anemia (AA). CD4+ T cells are divided into Th1 cells producing hematopoietic inhibitory cytokines like interferon-gamma and Th2 cells producing interleukin-4. We investigated the Th1/Th2 cell ratio in the peripheral blood of AA patients treated with immunosuppressive therapy (IST). There were 10 patients who responded well to IST (responders) and 3 patients who were refractory to IST (non responders). Th1 cells were lower in responders than in non-responders (16.2+/ 2.4% vs. 28.8+/-5.5%, respectively, p<0.05), whereas Th2 cells did not differ. The Th1/Th2 ratio was also significantly lower in responders than in non responders, being 13.2+/-1.5 and 40.4+/-5.1 (p<0.001), respectively. In three responders, the Th1/Th2 ratio was declined according to the hematological recovery (from 10.6 to 8.3, 16.3 to 10.9 and 11.8 to 9.5). Our results suggest that Th1 lymphocytes are more predominant in AA, and it may be very useful to monitor the Th1/Th2 ratio during IST. PMID- 15344556 TI - [Study of monoclonal gammopathy in Kagawa Prefectural Central Hospital]. AB - We investigated 361 patients with monoclonal gammopathy in whom immunoelectrophoresis was performed (1,037 tests) between 1986 and 2002 at Kagawa Prefectural Central Hospital. In this study, we identified 222 patients with monoclonal gammopathy of undetermined significance (MGUS). Malignant transformation of MGUS to multiple myeloma occurred in 15 patients (6.8%). No significant differences were observed in the means of total protein (TP), albumin (Alb), albumin/globulin ratio (A/G ratio), IgG, IgA, or IgM level in the initial examination between the patients who remained as MGUS and patients with malignant transformation of MGUS. However, the rate of progression to malignancy was high when the levels of normal immunoglobulins other than M protein were below the normal range. Since the number of MGUS cases detected and the number of protein fractionation performed were proportionate, and MGUS was found by protein fractionation in routine tests, protein fractionation is essential for detection of MGUS, and it is necessary to add serum protein fractionation to routine initial examination. In addition, long-term follow-up of patients with monoclonal gammopathy and preparation of a database of patient information are useful for monitoring the outcome. PMID- 15344557 TI - [Seasonal change of Vibrio vulnificus in the slime of tidal flats, seawater, and fishes collected along Ariake Sea, Japan]. AB - To investigate the seasonal change of Vibrio vulnificus in Ariake Sea, Japan, we attempted to isolate V. vulnificus from the slime of tidal flats, seawater, and fishes obtained from three harbors along Ariake Sea. The sample were collected twice a month from January to December, 2001. Also, we determined the biological characteristics of the individual isolates. V. vulnificus were isolated throughout the year, but the isolation ratios were higher during the warmer season from June to October. The isolates in the warmer season were able to grow in culture media containing 0.5% NaCl, whereas those isolated in the remaining seasons could not. Moreover, the isolates in the warmer season showed a greater hemolytic activity than those isolated in the remaining seasons. These results suggest that V. vulnificus isolated in warmer seasons are more vigorous in nature than those isolated in the remaining seasons. PMID- 15344558 TI - [Evaluation of autonomic dysfunction by novel methods]. AB - The autonomic nervous system innervates every organ in the body. Since autonomic disturbances affect patient survival, an understanding and recognition of these disturbances are important. We adopted several new methods to evaluate autonomic function accurately. 123I-metaiodobenzylguanidine scintigraphy can assess the cardiac autonomic function even in the presence of cardiac arrhythmia. Laser Doppler flowmetry, ultrasonographic study in the vessels and near-infrared spectrophotoscopy techniques serve as useful markers for screening the dysfunction of vasomotor neurons and blood circulation. Electrogastrography and the circadian rhythms of protein C secretion could be markers of the visceromotor nerves in the abdomen. Electrogastrography is a particularly useful tool for reflecting on functional changes in gastrointestinal motility. The evaluation of anemia could be a marker of autonomic dysfunction in the kidney and bone marrow in patients with familial amyloidotic polyneuropathy, pandysautonomia, and multiple system atrophy. Normocytic and normochromic anemia correlated with the severity of autonomic dysfunction were shown in these patients. We also evaluated the dysfunction of the neuroendocrine system and sudomotor neuron using our new autonomic function tests. The glucose-tolerance test could become one of the most useful clinical tools for detecting autonomic dysfunction in the endocrine system. Microhydrography and thermography could be useful tools for diagnosing the lesion site of dyshidrosis. Moreover, it is clinically important to check the systemic circulation and autonomic function in patients treated with sildenafil citrate and organ transplantation to save their lives. Our new autonomic function tests, such as laser-Doppler flowmetry and 123I-metaiodobenzylguanidine scintigraphy, are crucial tools in supplying the best symptomatic treatment for such patients. PMID- 15344559 TI - [Determination of biological substances using bioluminescent reaction based on luciferin-luciferase]. AB - Firefly luciferase catalyzes the oxidation of luciferin in the presence of ATP, magnesium ions and molecular oxygen, with a high quantum yield. Due to its high sensitivity and specificity for ATP, luciferase has been used for bioluminescent detection of ATP in various biological samples. However, it has not been well determined how to apply it in immunoassay. In this article, the use of various enzymes as labels in the design and development of immunoassays, and the combination of PCR/novel bioluminescent pyrophosphate assay, for detecting biomolecules is reviewed. PMID- 15344560 TI - [Clinics in prostate cancer]. AB - The prostate gland is characterized by the age-dependent development of abnormal proliferative disease, i.e., benign prostate hyperplasia and prostate cancer. Prostate cancer is the most important disease in our country among the elderly population. The diagnosis and treatment of prostate cancer have been dramatically improved due to the development of PSA screening, transrectal ultrasonography and new anti-androgen drugs, such as LH-RH analog, with fewer side effects. In this article, the pathogenesis, diagnosis and treatment of prostate cancer are summarized. PMID- 15344561 TI - [Advancements in PSA-based screening for prostate cancer]. AB - As the most frequently diagnosed cancer and the second leading cause of cancer death in most Western countries, prostate cancer represents a significant health care problem. The introduction of routine prostate-specific antigen (PSA) screening for asymptomatic men is still controversial. To solve uncertainties regarding the screening for prostate cancer, prospective randomized controlled trials are ongoing in the USA and Europe. The development of an optimal screening system may be one of the most important issues for screening for prostate cancer, and it should be set not only for reducing the mortality rate of prostate cancer, but also for reducing the cost. The best screening modality for the 1st step of mass screening for prostate cancer is the PSA test. Furthermore, the cut-off value should be set in an age-specific manner. The risk of developing prostate cancer in men with PSA levels of 4.0 ng/ml or lower increases when the baseline PSA levels are higher. Therefore, re-screening for men without suspicious findings for prostate cancer at the 1st step of screening should be set relative to the baseline PSA and digital rectal examination status. In the 2nd step of screening, the PSA density adjusted by the transition zone volume (PSATZD) and free/total PSA ratio (%f-PSA) may be useful in the selecting patients who should be biopsied. The optimal cut-offs for PSATZD and %f-PSA have not been confirmed, however, and an age-adjusted setting should be considered to detect clinically significant cancer. The method of prostate biopsy is also very important for improving the diagnostic accuracy for prostate cancer. The number of biopsy cores should be set relative to prostate volume. Furthermore, the clinically significant tumor volume may be smaller in younger men than in older men. Therefore, the optimal number of biopsy cores should be set according to age and prostate volume. Both an optimal screening system and minimally invasive treatments will be available in the future, and screening for prostate cancer may be more useful for elderly males. PMID- 15344562 TI - [Progress in standardization of total PSA immunoassays]. AB - The standardization of the total PSA assay was considered internationally after 1992, when the first Stanford Conference was organized by Prof. Dr. T. Stamey (Stanford University). In Japan, an analytical survey was carried out three times in the past six years by the committee of PSA standardization. The discrepancy in PSA values among each assay system was determined and the cause of discrepancy was studied. As a result, the principal cause of each discrepancy between kits was revealed, and 34% of all kits in the 1997 survey, 73% in the 2000 survey and 84% in 2003 were classified as equimolar response kits. The inter-kit variability of total PSA assay value was improved drastically. The standardization of the PSA assay has made great progress based on the establishment of purification of PSA from the seminal fluid pool and the guideline of value assignment approved by the NCCLS. In Japan, the special committee of PSA standardization was formed by the JCCLA in 2002. In this committee, we are considering the establishment of a reference measurement system including serum-based reference material, which is inactivated protease inhibitor, due to the traceability of the PSA concentration from the primary reference material that was certified by the WHO for patient serum. PMID- 15344563 TI - [Evaluation of the withdrawal of ACE inhibitors in coronary artery surgery]. AB - During cardiac surgery, as a result of surgical aggression, myocardial ischaemia and cardiopulmonary bypass, the renin-angiotensin-aldosterone mechanism is intensely activated. Our aim is to document whether, in the case of patients undergoing chronic treatment with lisinopril, the non-withdrawal of this inhibitor's administration before cardiac surgery and the administering of a last dose on the day of the operation are associated with coronary haemodynamic alterations. A study was made of 18 patients submitted to myocardial revascularization under extracorporeal circulation and distributed in two groups: group A) without ACE inhibitorsplacebo, group B) with ACE inhibitors (Lisinopril). Coronary blood flow (CBF) was determined by inverted thermodilution via Baim's catheter. Coronary and metabolic haemodynamic values were calculated. Lisinopril had no significant influence on the CBF or on the other above mentioned values. Therefore, it is not necessary to withdraw ACE inhibitors in cardiac surgery interventions. PMID- 15344564 TI - [Quality of life in diabetes]. AB - Different clinical features of diabetic patients and type of complications are certainly a critical components of the global individual perception of quality of life (qol); but also personal socio-cultural characteristics interfere concurrently. Qol in diabetes was assessed considering disease grade, complications and level of instruction in a population of 100 diabetic patients (30 m, 70 f), aged 66.99 +/- 13.68 years. Two questionnaires were administered to all patients; Sickness Impact Profile (SIP) and Functional Living Index (FLI). SIP is an index of psycho-social, physical and motor functionality; FLI derives from a scale devised for cancer patients and adapted to diabetic patients. Both were analysed by Spearman correlation test, and assayed vs. sociocultural profile and clinical symptoms. Neuro-muscular ailments were more prevalent in women; men experience severe limitations of working capacities an relational possibilities, with severe discomfort. In the whole, higher scores of SIP (greater disability p: ns) and lower score for FLI (scant well-being perception r = 0.29: p<0.01) are related with lower school instruction level. Global QOL score is related as well with degree of instruction (r = 0.22: p<0.03). So QOL is altered during chronic diseases: however, in diabetes, qol impairment does not seem related with severity, treatment features and complications of diabetes. Socio-cultural elements, and particularly instruction level quantified as school grades achieved, interfere with the manner of living diabetic disease. PMID- 15344565 TI - Psychiatric emergencies and services availability: a comparison of two hospitals in Rome. AB - The development of Psychiatric Emergency Services (PES) as an autonomous organization has allowed the registration of urgent request in different urban areas and their correlation with real clinical needs. Our aim was to investigate different patterns of psychiatric emergencies, considering that A) qualitative differences in diagnostic distribution could emerge in relation to the availability of local hospital services; B) gender differences could condition the type of demand. In particular, the availability of services appears to have a modulating effect on the demand: it's possible to deduce that socio-economic factors can influence demand and change over time the epidemiological features of patients availing of PES. PMID- 15344566 TI - [Management of neuropathic bladder in multiple sclerosis]. AB - It is estimated that almost 70% of patients affected by multiple sclerosis (MS) suffer from urinary symptoms, with devastant impact on Quality of Life (QoL). The major aims of management should be to ameliorate the patients quality of life and to prevent the frequent complications of bladder dysfunction such as infention and renal damage. Therapy can usually eliminate or reduce the symptoms of neuropathic bladder. In the following pages is discussed the complex management of urinary symptoms in MS patients. PMID- 15344567 TI - The pathogenesis of hepatocellular carcinoma is multifactorial event. Novel immunological treatment in prospect. AB - PURPOSE: To discuss exhanstively: complex molecular and cellular mechanism in Hepatocellular Carcinoma (HCC); effect of chronic inflammation and cirrhosis, accompained by regenerative process, on the development of HCC; genetic instability of liver cells of regenerating nodules; the relative role of hepatitis C virus (HCV) and hepatitis B virus (HBV) in hepatocarcinogenesis; tumorigenicity of aflatoxin B1 (AFB1); gene expression profiles in HCC; liver tumors and host defense; future perspectives of HCC treatment. DESIGN: We reviewed the most important studies on HCV. RESULTS: HCC is an aggressive malignancy with poor prognosis and is one of the most common tumor in the world. In the majority of cases, HCC is found in conjunction with cirrhosis of the liver. Chronic inflammation and cirrhosis, accompagnied by regenerative process, function as a tumor promoter, providing a common pathway from chronic HBV or HCV infection to HCC. The direct etiologic role of HBV and HCV for HCC is obscure. Tumor progression may be brought about in HCC by mutation of the p53 tumor suppressor gene. The prevalences of p53 mutations is similar in HBV-associated and HCV-associated HCCs. Another mechanisms of host defense are the production of transforming growth factor beta1 (TGFbeta1), and the induction of cytotoxic T lymphocytes; the failure of there mechanisms permits the process of hepatocarcinogenesis. Treatment with alpha interferon of chronic hepatitis is necessary to delary or prevent the progression to liver cirrhosis and development of HCC. Various therapies, such radical operation, intra-arterial chemoembolization, percutaneous intratumoral ethanol injection, radio-frequency ablation, have been employed, but there is still non satisfactory treatment. Recent advances in recombinant and gene delivery thechnologies suggest that gene therapy may be a promising alternative to explore. Furthemore, immunotherapy may become a modality for patients with HCC. Clinical application of vaccine immunotherapy with NY-ESO-1 derived peptides in HLA-A2 positive HCC patients will be possible. PMID- 15344568 TI - [Procedures and bioethics I]. PMID- 15344569 TI - [Erdheim-Chester disease: a non-Langerhans cell histiocytosis. A clinical-case and review of the literature]. AB - We make a retrospective evaluation of clinical and radiologic features, treatment, and outcome of Erdheim-Chester disease, a rare non-Langerhans cell histiocytosis. We report a case of Erdheim-Chester disease and review 60 cases from the literature. These cases are consider to have Erdheim-Chester disease when they have either typical bone radiographs (symmetrical long bones osteosclerosis) and/or histologic criteria disclosing histiocytic infiltration with distinctive immunohistochemical phenotype of the non-Langerhans cell histiocytes with positive staining for CD68 and negative staining for S-100 protein and CD1a. Our patient undergoes chemiotherapy according to the LCH-II stratification and therapy plan (Vinblastine, Etoposide and Prednisone) and thereafter receives Carboplatin and Etoposide, and Somatostatin. She is alive and clinically well 33 months after onset of symptoms and the lesions don't appear to progress at imaging examinations. In conclusion, Erdheim-Chester disease may be confused with Langerhans cell histiocytosis as it sometimes shares the same clinical (exophthalmos, diabetes insipidus) or radiologic (osteolytic lesions) findings. However, the characteristics radiological pattern of Erdheim-Chester disease together the immunohistochemical phenotype of hystiocytic infiltration supports the theory that Erdheim-Chester disease is a unique disease entity distinct. PMID- 15344570 TI - [Microhematuria]. PMID- 15344571 TI - [Diet and chronic corticosteroid therapy]. AB - Corticosteroid therapy is widely used in the acute and chronic treatment of different diseases, with consequent possible onset of typical side effects on multiple systems of the organism, including also energy metabolism and metabolism of water and minerals (sodium, potassium, calcium and phosphorus). Clinical signs, related to the type and dosage of the steroidal drug, may lead to secondary illnesses with variable degrees of severity, depending on proneness of the individual patient and on the underlying disease that motivated the treatment. The role of dietetic intervention in the management of a patient chronically receiving corticosteroid therapy is not ancillary, although often underestimated, its aim being the reduction of some long-term therapy related side effects, and the correction of major metabolic derangements. In particular, the diet shall be moderately rich in protein (1.5 g/Kg/day of proteins) and low in fat (< 30% of calories, obtained mostly from unsaturated fatty acids), based mainly on complex carbohydrates (80%), providing 50% of the caloric intake. Diet has to be specifically characterized by food containing little sodium and yielding high amounts of water, calcium, magnesium and potassium. Minor directions concern the reduced intake of ethanol and purines. The efficacy of supplementation with antioxidant vitamins (vitamin C and E) and selenium is currently under investigation. PMID- 15344573 TI - Piracy and other dastardly deeds. PMID- 15344572 TI - [Ascites]. PMID- 15344574 TI - Avascular necrosis of the femoral head: etiology, pathophysiology, classification, and current treatment guidelines. PMID- 15344575 TI - Chronic exertional compartment syndrome. AB - Chronic exertional compartment syndrome is an overuse condition affecting primarily active, athletic people. Its etiology is unclear, but several theories have been proposed. The syndrome is characterized by recurrent, often severe muscle compartment pain that occurs with vigorous exercise and subsides with rest. Physical examination usually provides little helpful information. Compartment pressure measurement before and after exercise is the mainstay of objective diagnosis. Conservative management has been highly unsuccessful. Overall, considerable success can be expected from fasciotomy of the involved compartments followed by a rigorous rehabilitation program, though recurrence of symptoms is a reported complication. Recent research efforts have focused on the development of less invasive and more accurate diagnostic modalities and safer and more effective surgical procedures. PMID- 15344576 TI - Immune response in patients receiving fresh osteochondral allografts. AB - Immunologic response to cartilage-specific protein (CSP) antigens was tested in patients treated with fresh osteochondral allografts. Eight of 14 allograft recipients showed immune reactivity against CSP, whereas only 2 of 14 controls were reactive (P<.05). The molecular weights of the most common immunoreactive proteins were 220 kd and 95 kd. The precise identity of these proteins could not be determined using standard anticollagen antibodies. These data indicate the presence of antibodies to CSP--which supports the concept of immunologic response to the cartilage component of osteochondral allografts. PMID- 15344577 TI - Cat-scratch disease of the upper limb with ulnar nerve involvement. PMID- 15344578 TI - Reconstructive operations for the upper limb after brachial plexus palsy. AB - Limited function due to paralysis following brachial plexus lesions can be improved by secondary operations of the bony and soft tissue. Between April 1994 and December 2000, 109 patients suffering from arm-plexus lesions underwent a total of 144 reconstructive operations guided by our concept of integrated therapy. The average age at the time of surgery was 32 years (range: 15-59). The following operations were performed: shoulder arthrodesis (23), trapezius transfer (74), rotation osteotomy of humerus (9), triceps to biceps transposition (9), transposition of forearm flexors or extensors (8), latissimus transfer (7), pectoralis transfer (1), teres major transfer (1), transposition of flexor carpi ulnaris to the tendons of extensor digitorum (10), and wrist arthrodesis (2). Prospectively, in all patients, the grade of muscle power of the affected upper extremity was evaluated prior to surgery. The follow-up period for all 144 operations was, on average, 22 months (range: 6-74). By means of operative measures, almost all patients obtained an improvement of shoulder function (100%) and stability (>90%), elbow flexion (85%), and hand, finger, and thumb (100%). When muscles malfunction after brachial plexus lesions, one should take into account the individual neuromuscular defect, passive joint function, and bony deformities; different procedures such as muscle transpositions, arthrodeses, and corrective osteotomies can then be performed to improve function of the upper extremity. Each form of operative treatment presents patients with certain benefits and all are integrated into a total treatment plan for the affected extremity. PMID- 15344579 TI - Freehand locking of intramedullary nail. AB - Placing a loop of suture on an intramedullary nail locking screw keeps the screw firmly on the screwdriver. The screw can be retrieved should the need arise. PMID- 15344580 TI - Practical procedures & aesthetic dentistry. PMID- 15344581 TI - Sensor-based intraoral digital radiography. PMID- 15344582 TI - Simplified ceramic restorations using CAD/CAM technologies. AB - The advent of CAD/CAM systems that enable computerized, in-office fabrication of porcelain restorations has revolutionized the delivery of precise, durable, and aesthetic restorations. Using specialized software programs that facilitate visualization of the desired restoration on the model prior to actual fabrication and scanning devices that can be applied to the intraoral structures have further improved the clinician's ability to determine the necessary restorative parameters that will ensure an accurate fit. This article presents the fabrication and delivery protocol used for CAD/CAM-fabricated restorations. PMID- 15344583 TI - Aesthetic implant management of multiple adjacent failing anterior maxillary teeth. PMID- 15344584 TI - Replacing a single molar with dental implants. PMID- 15344585 TI - Improving adhesion: total-etch versus self-etch techniques. PMID- 15344586 TI - Identification and correction of common impression concerns: protocol and considerations. AB - Precise impression-making techniques are critical to indirect restorative success. By ensuring that a concise protocol is implemented during the impression capture phase, clinicians can predictably deliver optimal results. Common complications that may arise during impression making include inaccurate marginal detail, bubbles, tears, pits, and voids. Proceeding with impression making when such damage is evident will reduce the accuracy of restorative fit and integrity. This article discusses potential concerns during impression capture and addresses approaches to improve the overall quality of impressions taken. PMID- 15344587 TI - Contemporary curing light technology. PMID- 15344588 TI - Pickup impression technique for fabricating the surgical template for implant placement and provisional restoration. PMID- 15344589 TI - Prophylactic root resection and periapical grafting for anterior implant aethetics: an integration of tissue-reconstruction and tissue-preservation concepts. AB - The presence of active infections involving the labial bony plate of a failing maxillary anterior tooth presents a challenging situation for aesthetic implant replacement. Not only would the septic state of an abscess be detrimental for immediate implant placement and bone augmentation, but the accompanying bony destruction would also increase the likelihood and magnitude of gingival recession, making this a bioaesthetic challenge. This article describes a technique that integrates tissue-reconstruction and tissue-preservation concepts to achieve peri-implant tissue aesthetics as well as its clinical rationale. PMID- 15344590 TI - A working classification of scleroderma spectrum disorders: a proposal and the results of testing on a sample of patients. AB - OBJECTIVE: To propose a working classification for scleroderma spectrum disorders (SDS), based on criteria shown in the past to be specific for the condition: sclerodermatous skin involvement, specific autoantibodies and specific microvascular abnormalities, to obtain mutually exclusive subgroups of SDS and to test this classification on a sample of patients. This is an attempt to improve the current classifications used in research studies, which are all based on the extent of skin involvement only. METHODS: Patients (n= 165) referred to the Microvascular Laboratory of the Division of Rheumatology between 1976 and 1992, who had participated in various other studies in the Division and for whom therefore systematically collected clinical and laboratory data were available. Evaluation of skin involvement, clinical, laboratory, and microvascular studies were performed independently and a subgroup assignment was made "blindly" from coded data. RESULTS: Results demonstrated that significant differences were present between our subgroups of SDS with regard to meaningful variables. Comparisons of our results with previously used classifications such as the ACR preliminary criteria study and later subsets used by more recent investigators, demonstrated that our classification permitted us to include less severe cases than those meeting ACR criteria. CONCLUSION: Our results show that by dividing SDS into groups using the cutaneous, microvascular and autoantibody characteristics, while using well-defined criteria and inclusion-exclusion rules to obtain mutually exclusive subgroups, can reveal differences that may otherwise be masked. This study has suggested new avenues of research. A replication study should be performed with a systematic follow-up. This is a working classification, i.e. it can be modified as new findings using updated technology become available. The focus should be on clearly defining criteria for classification and collected data. Then between-center comparisons will be possible. PMID- 15344591 TI - Undifferentiated connective tissue diseases in 2004. AB - The existence of patients with signs and symptoms suggestive of a systemic autoimmune disease but not fulfilling the classification criteria for defined diseases is a common experience in clinical practice. The first description of these diseases dates back to 1980, when LeRoy proposed the term "Undifferentiated Connective Tissue Syndromes" (UCTS) to define the early phases of connective tissue diseases that at their onset are undefined, as opposed to mixed or overlapping syndromes in which the disease appears well defined. LeRoy also raised the fundamental question as to whether a proportion of patients with an undifferentiated profile may see their condition remain undifferentiated or even experience a remission of all pathologic features, rather than evolving to a definite connective tissue disease (CTD). Many studies conducted on undifferentiated diseases have shown that up to 75% of patients will not develop a defined CTD and that these conditions exhibit typical clinical and serological manifestations and a good prognosis. We and others have defined these conditions as Undifferentiated Connective Tissue Diseases (UCTD) and it is our opinion that the UCTD represent distinct clinical entites. UCTD could offer an ideal clinical model for the study of single autoantibody specificities, the effects of various factors (such as pregnancy) on the disease course, and the general pathogenesis of autoimmune conditions. Before they can be so used, more must be discovered about their nature and characteristics, and studies will be necessary to improve the sensitivity and specificity of the existing preliminary classification criteria. PMID- 15344592 TI - Vascular involvement in systemic sclerosis (SSc). AB - The vascular endothelium came into view almost three decades ago following the introduction of the vascular hypothesis in scleroderma pathogenesis by Dr LeRoy. Since that time, the endothelial cells, and other vascular cells, became the focus of investigations aimed at eludicating the etiology, pathogenesis and treatment of scleroderma. This review will summarize Dr LeRoy's commitment to the disease, the relevant progress made since the introduction of the vascular hypothesis, and what we have learned since then about the vascular disease in scleroderma. PMID- 15344593 TI - Regulation of substance P mRNA expression in human dermal microvascular endothelial cells. AB - Vascular endothelial cells have been identified as a source of substance P (SP) which may act in an autocrine/paracrine fashion to bring about nitric oxide (NO) dependent vasodilatation and mitogen-induced cell division or immunologic and inflammatory responses. Whilst SP is localised in and released from endothelial cells, an endothelial mRNA expression of SP has not previously been shown. In the present study, mRNA expression of SP in human dermal microvascular endothelial cells is demonstrated using in situ hybridisation techniques with enhancement procedures. Incubation of microvascular endothelial cells with nerve growth factor (NGF) under conditions of increased shear stress increases the mRNA expression and release of SP Endothelin (ET) release is also enhanced. These changes are pertinent to circulatory, events that may occur in Raynaud's phenomenon in systemic sclerosis. PMID- 15344594 TI - Gamma/delta T cells in placenta and skin: their different functions may support the paradigm of microchimerism in systemic sclerosis. AB - Increased amounts of foetal cells persisting after pregnancy could be involved in the pathogenesis of systemic sclerosis (SSc) and other autoimmune diseases. Evidence suggests a specific role for a subset of T lymphocytes showing the gamma/delta T cell receptor (TCR) at the foetal/maternal interface. gamma/delta T cells significantly increase in the early pregnancy decidua and recognize trophoblast antigens, probably a highly evolutionarily conserved molecule such as Hsp60 or Hsp60-derived peptides, and are likely to suppress the maternal anti foetal immune response via TGFbeta production, thus contributing to pregnancy maintenance. The similarity between the presence of host gamma/delta T cells in pregnancy decidua and in SSc skin suggests that the functional activities of these cells can be differentially modulated by several mechanisms including the nature of the antigen and the involved organs. To support pregnancy, the decidual microenvironment might induce a Th2 activity of host Vdelta1 + T cells. On the contrary, either the presence of foetal cells in the skin of SSc patients or an as yet unidentified stimulus (i.e. infections), may trigger Vdelta1 + T cells toward the Th1 phenotype with the subsequent activation of cytotoxicity and modulation of the cytotoxic alpha/beta acquired T cell response. On these grounds, understanding the mechanisms which prevent the maternal immune system from rejecting a semiallogenic foetus could be helpful to understand the development of some autoimmune diseases, and potentially to develop new targeted therapeutic strategies. PMID- 15344595 TI - The 72-kilodalton IE-1 protein of human cytomegalovirus (HCMV) is a potent inducer of connective tissue growth factor (CTGF) in human dermal fibroblasts. AB - Latent human cytomegalovirus (HCMV) infection has been implicated in diseases characterized by tissue remodeling. Because of recent evidence indicating the possibility of a partial HCMV reactivation, the purpose of this study was to examine the role of the HCMV immediate early (IE) genes in the regulation of extracellular matrix (ECM) related host genes. Adenoviral vector expressing IE1 was generated to allow efficient gene delivery into human fibroblasts. IE1 stimulated the prolonged expression of connective tissue growth factor (CTGF) and TIMP1. IE1-dependent stimulation of CTGF was partially mediated by TGF-beta. Moreover, whereas collagenous proteins and collagen type 1 mRNA were only transiently induced by IE1 in the majority of fibroblasts, in selected fibroblast strains IE1 induced persistent ECM upregulation for up to 120 hours. This study suggests that transient or limited HCMV reactivation may play a direct role in abnormal matrix remodeling in GVHD, scleroderma, atherosclerosis and other HCMV linked diseases. PMID- 15344596 TI - Immunoglobulin GM genes and IgG antibodies to cytomegalovirus in patients with systemic sclerosis. AB - OBJECTIVE: To determine the role of GM and KM genes--genetic markers of immunoglobulin gamma and kappa chains, respectively--in humoral immunity to human cytomegalovirus (HCMV) in patients with systemic sclerosis (SSc; scleroderma). METHODS: A total of 137 Caucasian patients with SSc and 145 ethnically matched controls were genotyped for GM f/3, z/17, n+/23+, n-/23-, KM 1, and KM 3 alleles by polymerase chain reaction-restriction fragment length polymorphism and direct DNA sequencing methods. IgG antibodies to HCMV were measured by an enzyme-linked immunosorbent assay. RESULTS: In SSc patients, GM f,z genotypes were strongly associated with the occurrence of anti-HCMV IgG antibodies. The frequency of the GM f homozygotes was lower (42.2 vs 62.2%; p= 0.02; OR=0.4) and the frequency of the GM f,z heterozygotes was higher (51.1 vs 26.7%; p = 0.006; OR = 2.8) in SSc patients with IgG antibodies to HCMV than in subjects who lacked these antibodies. This association was not observed in the control group. KM and GM n genotypes were not significantly associated with the prevalence of these antibodies. CONCLUSION: GM f,z alleles or alleles in linkage disequilibrium with them influence the generation of IgG antibodies to HCMV in patients with SSc. PMID- 15344597 TI - Thrombin-mediated cellular events in pulmonary fibrosis associated with systemic sclerosis (scleroderma). AB - The vascular hypothesis for the pathogenesis of systemic sclerosis was perhaps Professor LeRoy's most important scientific contribution. One early and important consequence of vascular injury is the release of activated thrombin. In this manuscript we present our data and review the current understanding of the role played by thrombin in the process of fibrosis, particularly as it relates to scleroderma lung disease. Thrombin's cellular effects are intimately involved in promoting myofibroblast differentiation, endothelial cell activation, extracellular matrix protein deposition, and the induction of important profibrotic factors. Such studies confirm that thrombin is one of the major mediators in the development and progression of pulmonary fibrosis. Therefore, targeting the major receptor of thrombin, PAR-I, and its downstream signaling molecules may lead to novel therapeutic approaches for the management of scleroderma lung fibrosis. We are indebted to Dr LeRoy for his many contributions to the field of scleroderma, and for all that he did to stimulate our interest in these studies. PMID- 15344598 TI - Global expression analysis of the fibroblast transcriptional response to TGFbeta. AB - OBJECTIVES: Transforming Growth Factor-beta (TGFbeta) is the predominant cytokine in all forms of fibrotic reactions. As well as being secreted by immune modulators of fibrosis such as macrophages, it is involved in an autocrine feedback loop of fibroblast stimulation whose regulation is still poorly understood. We wished to gain some insight into the mechanisms of the fibroblast response to TGFbeta. METHODS: We undertook an exhaustive transcript profiling experiment using a widely validated restriction enzyme based method for identifying differentially expressed genes (GeneCalling). Transcriptional responses throughout a 24-hour time course were examined at multiple time points and classified. RESULTS: By 24 hours of TGF treatment over 1000 bands, representing a large number of transcripts, were down- or upregulated greater than 2-fold. All of the known genes responsive to TGFbeta, such as collagen and connective tissue growth factor, were upregulated. CONCLUSIONS: This encyclopedic method revealed many unknown transcriptional responses to TGFbeta including the upregulation of a variety of less expected cytoskeletal and matrix components, as well as interactions between the TGFbeta and tumor necrosis factor (TNF) pathways and alterations in cell death-related pathways. These may in part explain the idiosyncratic responses of mesenchymal cells to TGFbeta. PMID- 15344599 TI - What did we learn by studying scleroderma fibroblasts? AB - Systemic sclerosis (SSc, scleroderma) is a complex disease manifesting itself by fibrosis of skin and other internal organs. Fibroblasts isolated from SSc lesions and cultured in vitro are characterized by the increased synthesis of collagen and other extracellular matrix (ECM) proteins, consistent with the disease phenotype. Cultured SSc fibroblasts therefore serve as a principal experimental model for studying the molecular and cellular mechanisms involved in collagen overproduction in this disease. This article provides an overview of transcription factors that are deregulated in SSc fibroblasts. The possible origin of SSc fibroblasts is also discussed. PMID- 15344600 TI - What's wrong with the scleroderma fibroblast? PMID- 15344601 TI - Scleroderma fibroblasts constitutively express the long pentraxin PTX3. AB - OBJECTIVE: PTX3 is a secreted molecule which consists of a C-terminal domain similar to classical pentraxins (e.g. C-reactive protein) and of an unrelated N terminal domain. Unlike the classical pentraxins, PTX3 is expressed in response to IL-1beta and TNF-alpha but not to IL-6. The present study was designed to investigate the expression of PTX3 in normal and scleroderma fibroblasts. METHODS: Normal and SSc fibroblasts were cultured in the presence and absence of inflammatory cytokines. PTX3 mRNA expression in fibroblasts was evaluated by Northern analysis. PTX3 protein levels in fibroblast culture medium were estimated by ELISA. RESULTS: Normal fibroblasts were induced to express high levels of P7X3 mRNA by IL-1beta and TNF-alpha but not by other cytokines or growth factors. Scleroderma fibroblasts, unlike normal fibroblasts, constitutively expressed high levels of PTX3 in the absence of deliberate stimulation. The constitutive expression of PTX3 in SSc fibroblasts was not modified by anti-TNF-alpha antibodies or IL-1 receptor antagonist. In contrast, IFN-gamma and TGF-beta inhibited the constitutive but not the stimulated expression of PTX3 in SSc fibroblasts. CONCLUSIONS: PTX3 is a main feature of activated scleroderma fibroblasts. PMID- 15344602 TI - Regulation of connective tissue homeostasis in the skin by mechanical forces. AB - Mechanical forces come in a variety of different forms and act on many more tissues than the obvious vascular and musculo-skeletal systems. Next to soluble mediators, they have caught our attention as potent regulatory parameters in modifying the metabolism and phenotype of cells. This paper concentrates on the response of the skin to tensile forces and describes the adaptive phenotype of fibroblasts residing in the dermal connective tissue. Work from a number of different groups suggests that tension induces in these cells many biological properties which have been found in pathological conditions, e.g. in fibrotic skin lesions. Mechanical tension can therefore be regarded as an additional important regulatory parameter, which we would like to better understand with respect to mechanosensing cellular structures and specific or shared signaling pathways. PMID- 15344603 TI - Transcription activity in fibroblasts from the adult tight skin (TSK) mouse. AB - OBJECTIVE: To assess if adult tight-skin mouse (TSK) skin fibroblasts have a parallel increase in transcription of collagen and non-collagen genes. METHODS: One-year-old TSK/+ and +/+ (normal littermate) dermal fibroblasts were transfected (lipotransfection) with plasmid constructs containing Chloramphenicol Acetyl Transferse (CAT) gene, directed by promoters of mouse alpha1(I) alpha2(I) and alpha1(III) collagen genes, and by viral enhancers of Simian virus 40, Rous sarcoma virus and an LTR from a Syrian hamster tumour retrovirus. Syrian hamster derived tumour cell lines MF2B and GRI, and fibroblast cell line 3T3/NIH were used as controls. In some experiments, transfected cells were treated with hormones as transcription activating factors. Mixing experiments of tumour cells and TSK/+ or +/+ fibroblasts were done to study potential inhibitors. RESULTS: Collagen genes promoters failed to induce transcriptional activity in TSK/+ or +/+ fibroblasts, even in the presence of hormone treatment. Mixing experiments did not reveal inhibitor factors acting in these fibroblasts. Viral enhancers induced 2 to 5 times more transcription activity in TSK/+ than in +/+ fibroblasts. CONCLUSION: Increased transcription of viral enhancers and not of collagen genes in adult TSK fibroblasts, suggests the presence of transcription activating factors independent of collagen gene activation. PMID- 15344604 TI - The tight skin mouse: an animal model of systemic sclerosis. AB - The search for an animal model of systemic sclerosis (SSc) was tenaciously pursued by E.C. LeRoy. We studied several aspects of the tight skin mouse (Tsk) genetics and pathogenesis under his stimulating influence that contributed to a better understanding of the fibrotic scleroderma-like phenotype of this mouse. The identification of the fibrillin-1 mutation in the Tsk mouse and the characterization of the cellular and molecular pathways leading to Tsk fibrosis by numerous research groups has opened new avenues in the investigation of human SSc. The enigmatic connections between autoimmunity and ECM homeostasis in fibrotic diseases have received extensive attention in this mouse in which a prirmary alteration of a connective tissue microfibrilar protein leads to the reproduction of cellular and autoimmune abnormalities strikingly similar to human SSc. The use of this mouse as a tool to explore anti-fibrotic therapeutic interventions has demonstrated its value in providing useful information on the search for a therapy for this untreatable facet of human disease. PMID- 15344605 TI - Treatment of early diffuse cutaneous systemic sclerosis patients in Japan by low dose corticosteroids for skin involvement. AB - Systemic sclerosis (SSc) is a multiorgan systemic disorder characterized by excessive collagen deposition. This disease is usually classified into two clinical subsets - diffuse cutaneous SSc (dSSc) and limited cutaneous SSc (lSSc). We have proposed low-dose oral corticosteroid treatment with indication criteria consisting of the following three conditions: (1) early onset; (2) edematous changes; and (3) rapid progression. Cases satisfying 2 or 3 criteria have a definite indication, while those meeting 0 or 1 criterion have no indication. We treated 23 cases of dSSc by low-dose oral corticosteroid (prednisolone 20 mg/day as the initial dosage) and evaluated the effect using the modified Rodnan total skin thickness score (m Rodnan TSS). The mean initial TSS (20.3+/-9.3) decreased significantly to 12.8+/-7.0 after one year of treatment (p < 0.005) and to 8.7+/ 6.1 at final evaluation (p < 0.001). Thus, we confirmed the usefulness of oral corticosteroid treatment for early dSSc in Japanese patients. PMID- 15344606 TI - Guideline on the management of melanoma. AB - OBJECTIVE: 1. The Guideline for the Management of Melanoma has been developed in an attempt to improve management through the process of locating the best available evidence on which to base decisions. It is expected to help to improve the quality of care. 2. Melanoma remains a common cancer in South Africa. Despite the achievement of earlier diagnosis, it would appear from current statistics that at least 850 people continue to die of melanoma each year. Many of these deaths occur at a younger age than for other solid tumours, so the number of years of life lost due to melanoma exceeds that of many other cancers. It is seen as imperative to maximise effective management of melanoma. 3. Prevention of melanoma has not yet been achieved, and there are no conclusive data to show that current promotion of sun avoidance has substantially altered its incidence. 4. Early detection is an important factor in melanoma management, with diagnosis based mainly on changes in colour, diameter, elevation and border (irregularity of outline) of a skin lesion, asymmetry of a lesion, or a lesion different from other naevi. People at high risk of melanoma should be offered a surveillance programme. RECOMMENDATIONS: 1. All clinicians should be trained in the recognition of early melanoma. 2. If there is doubt about a lesion, the patient should be referred for specialist opinion (if readily available) or a biopsy should be undertaken. Biopsy of a pigmented lesion should be done only on the basis of suspicion of melanoma. Excision with a 2 mm margin is adequate. 3. Prophylactic excision of benign naevi is not recommended. In general, elective lymph node dissection is not indicated. 4. People with high-risk primary melanoma, lymph node involvement and melanoma in unusual sites (e.g. mucosal and disseminated melanoma) should be managed with support from a melanoma centre. VALIDATION: Melanoma management involves many medical specialties. Guidelines should therefore be developed through a multidisciplinary consensus. The Melanoma Advisory Board consists of a forum of dermatologists, oncologists, plastic surgeons and pathologists. PMID- 15344607 TI - Guideline for prophylactic anticoagulation. AB - INTRODUCTION: Prophylactic anticoagulation in South Africa is unfortunately under prescribed. This has led to unacceptable mortality and morbidity in numerous patients. METHOD: The Southern African Society of Thrombosis and Haemostasis held two meetings at which all the available literature as well as guidelines from other societies were reviewed. The specialties represented on the committees included Anaesthetics, Cardiology, Clinical Haematology, Critical Care, Gynaecology, Haematopathology, Internal Medicine, Neurology, Orthopaedic Surgery, Pulmonology and Vascular Surgery. A draft document was presented at both meetings, which was altered by consensus agreement. The guidelines were adjudicated by a recognised external international expert to avoid local bias. RESULTS AND CONCLUSION: A guideline for prophylaxis in medical and surgical patients has been produced for the South African situation. It is hoped that this will lead to improved anticoagulation practice in this country, which we believe will directly benefit patient outcome. PMID- 15344608 TI - Does an impaired clinician threaten your practice? Legal considerations. PMID- 15344609 TI - National health care improvements. PMID- 15344610 TI - Beware of 'most-favored-nation' clauses. PMID- 15344611 TI - The winds of change are blowing: get ready for ICD-10. PMID- 15344612 TI - Ockham's razor cuts confusion surrounding management. PMID- 15344613 TI - Web-based practice management system can streamline your operations. PMID- 15344614 TI - Using the process map to improve your bottom line. . . and maintain your sanity. PMID- 15344615 TI - Duck and cover. Medical liability insurance choices in today's market. PMID- 15344616 TI - Has HIPAA reduced your billings? Why your practice isn't seeing the same revenue it used to. PMID- 15344617 TI - Shell-shocked. Looking out for the well-being of physicians--particularly in this malpractice environment--is the mark of a professional medical practice manager. AB - Practice administrators often find that an unwritten part of their job descriptions is running interference when one of the group's physicians faces a malpractice lawsuit. Supporting the doctor in the work environment and encouraging open discussion about malpractice long before lawsuits occur is a key part of running an effective organization. PMID- 15344618 TI - When OSHA comes knock-knock-knocking. Seven rules to avoid the poorhouse. AB - What would happen if an inspector from the Occupational Safety and Health Administration (OSHA) visited your practice? The author provides guidelines for handling an OSHA inspection and describes the procedures of the federal agency and expectations for compliance. She also describes the types of violations and penalties that OSHA can impose on a medical practice. PMID- 15344619 TI - Stark II, phase II. Positive changes and lingering uncertainties. AB - The new physician self-referral rule--referred to as the Stark law--has key implications for medical groups. This second article of a two-part series covers important provisions and changes the government made in phase II, including a temporary noncompliance grace period, new procedures defined as designated health services, a narrow exception for professional courtesy and clarification of the academic medical center exception. Lingering uncertainties about the rule remain, however. PMID- 15344620 TI - Flying high. Alan J. Beason, FACMPE, MGMA member and chief executive officer, Cardiovascular Consultants LLP, Shreveport, La. PMID- 15344621 TI - Evidence and the life-long learner. PMID- 15344622 TI - Comparison of mineral trioxide aggregate and formocresol as pulp-capping agents in pulpotomized primary teeth. AB - PURPOSE: The aim of this study was to use clinical, radiographic, and histologic examinations to compare the relative success of gray mineral trioxide aggregate (MTA), white MTA, and formocresol as pulp dressings in pulpotomized primary teeth. METHODS: Twenty-four children, each with at least 3 primary molars requiring pulpotomy, were selected for this study's clinical and radiographic portion. An additional 15 carious primary teeth planned for serial extraction were selected for this study's histologic portion. All selected teeth were evenly divided into 3 test groups and treated with pulpotomies. Gray MTA was used as the pulp dressing for one third of the teeth, white MTA was the dressing for one third, and the remaining one third were treated with formocresol. The treated teeth selected for the clinical and radiographic evaluations were monitored periodically for 12 months. The treated teeth selected for histologic study were monitored periodically and extracted 6 months postoperatively. RESULTS: Four children with 12 pulpotomized teeth failed to return for any follow-up evaluations in the clinical and radiographic study. Of the remaining 60 teeth in 20 patients, 1 tooth (gray MTA) exfoliated normally and 6 teeth (4 white MTA and 2 formocresol) failed due to abscesses. The remaining 53 teeth appeared to be clinically and radiographically successful 12 months postoperatively. Pulp canal obliteration was a radiographic finding in 11 teeth treated with gray MTA and 1 tooth treated with white MTA. In the histologic study, both types of MTA successfully induced thick dentin bridge formation at the amputation sites, while formocresol induced thin, poorly calcified dentin. Teeth treated with gray MTA demonstrated pulp architecture nearest to normal pulp by preserving the odontoblastic layer and delicate fibrocellular matrix, yet few inflammatory cells or isolated calcified bodies were seen. Teeth treated with white MTA showed a denser fibrotic pattern, with more isolated calcifications in the pulp tissue along with secondary dentin formation. CONCLUSIONS: Gray MTA appears to be superior to white MTA and formocresol as a pulp dressing for pulpotomized primary teeth. PMID- 15344623 TI - Leukocyte esterase and protein levels in saliva, as indicators of gingival and periodontal diseases in children. AB - PURPOSE: This study was to determine if dip stick assays of children's saliva for leukocyte esterase or protein (Serim Research Corporation, Elkhart, Ind) reflect the presence or severity of gingival or periodontal diseases in children. METHODS: The study included 13 children with periodontitis [study group] and a control group of 17 children without periodontitis. The saliva leukocyte esterase and protein values (scales from 1 to 4) were tested with dip stick analyses. The gingival (GI) and plaque indices (PI) presence and number of sites with periodontitis, demographic data, systemic condition, caries prevalence, and the presence of dental restorations were recorded and their relationship to leukocyte esterase or protein values were analyzed. RESULTS: Most children had a protein value of 2 or 3 or a leukocyte esterase value of > or = 3. Significant differences in the distribution of protein values by the presence/absence of periodontitis (chi-square, P<0.001), or the number of sites with periodontitis by protein value (chi-square, P=.005; ANOVA, P=.03) were evident; No. 4 protein values were found only in children with periodontitis, and No. 2 and No. 3 protein values were mostly found in children without it. CONCLUSIONS: Dip stick protein analysis of saliva of children has the potential to differentiate children with periodontitis or with fewer periodontal lesions. PMID- 15344624 TI - Children's fear and behavior in private pediatric dentistry practices. AB - PURPOSE: This study assessed the proportion of children with dental fear, the proportion of children with negative behavior and the relationship of children's dental fear and children's negative behavior in private pediatric dentistry practices in western Washington state. METHODS: A total of 421 children seen in 21 private pediatric dentistry practices in western Washington State participated. The average age of the children was 6.8+/-2.8 years (range=0.8-12.8 years). An average of 21 children were studied per practice (range=7-25 children). Dental fear was measured using the parents' version of the Dental Subscale of the Child Fear Survey Schedule (CFS). Behavior of the child during treatment was rated using the Frankl scale. Additional data were collected regarding the child's previous experiences and parental fear. RESULTS: The children's average item score on the 15-item CFS was 2+/-0.7, corresponding to a total score of 29.6 out of 75, where 75 indicates maximum fear. The proportion of children with dental fear, defined as an average item score of > or = 2.5 (corresponding to a total score of > or = 38), was 20% (85/421, 95% CI=16.3, 24%). The proportion of children who displayed negative behavior during treatment was 21% (95% CI=17.5, 24%). A prevalence ratio of 2.4 was calculated to describe the relationship between children's behavior during treatment and children's dental fear prior to dental treatment. Multivariate logistical regression analysis showed children with dental fear, younger children, and children exposed to treatment involving local anesthesia have higher odds of displaying negative behavior. CONCLUSIONS: The proportion of children with dental fear in private pediatric dentistry practices was 20%, and the proportion of children with negative behavior during treatment was 21%. Children with negative behavior had greater odds of having dental fear and children with dental fear had greater odds of having negative behavior. Screening for dental fear may allow pediatric dentists to prepare children more adequately for positive treatment experiences. PMID- 15344625 TI - Sealing ability of new generation adhesive systems in primary teeth: an in vitro study. AB - PURPOSE: This study evaluated the sealing ability of different types of restorative-adhesive combinations on deciduous molars in vitro. METHODS: Facial and lingual Class V cavities were prepared in 120 primary teeth. They were randomly divided into 8 groups of N= 15, in which different adhesives were used (XE=Xeno III; LP=Adper Prompt L Pop; IB=I Bond; SB=Scotch Bond 1; EP=Etch & Prime 3.0; AS=AdheSE; OB=Optibond Solo plus self-etch primer; CS=Clearfil SE Bond). All cavities were restored with composite Z 250. After thermocycling and immersion in 2% methylene blue, the dye penetration was evaluated under a microscope. RESULTS: In enamel and in cementum: the best seals were obtained with XE and LP, followed by CS, AS, IB, OB, SB, and EP (P=.001). No significant differences were recorded in the microleakage degree between the cementum and the enamel margins (P=.40). CONCLUSIONS: In this in vitro model, Xeno III and Adper Prompt L Pop provided the best seals both at the enamel and the cementum margins of Class V cavities in primary molars. PMID- 15344626 TI - Survey of Oral Health Status of Maryland Schoolchildren, 2000--2001. AB - PURPOSE: The purpose of this study was to describe dental caries experience in the primary and permanent dentitions of Maryland schoolchildren in kindergarten and third grade, with an emphasis on untreated disease. METHODS: Data came from the Survey of the Oral Health Status of Maryland Schoolchildren, 2000--2001. Outcome variables were measures of dental caries experience for primary and permanent teeth. Dental caries was scored only for lesions that were >0.5 mm, measured with a periodontal probe. Radiographs were not used. Descriptor variables included grade level, gender, race/ethnicity, eligibility for free or reduced meals at school, parent/ guardian education, dental insurance status, and region. The statistical software program SUDAAN was used to produce weighted estimates and account for the complex sampling design. RESULTS: More than 2 in 10 schoolchildren had a history of dental caries. In children with a history of disease, approximately half of the experience was due to untreated decay. Controlling for confounders, disparities in untreated dental caries were significantly associated with region, race/ethnicity, and parent/guardian education. CONCLUSIONS: Success in meeting the dental caries treatment needs in children will require attention to multiple family-level, community-level, and system-level barriers, especially among low socioeconomic status populations. Unless barriers are overcome, dental caries disparities will continue in Maryland. PMID- 15344627 TI - Parental satisfaction with bonded resin composite strip crowns for primary incisors. AB - PURPOSE: The purpose of this study was to evaluate the parental satisfaction of bonded resin composite strip crowns for the treatment of maxillary anterior primary incisors and compare their satisfaction with the clinical evaluation and success of the crowns. METHODS: This was a retrospective, clinical study of patients who had strip crowns (SC) placed on maxillary primary incisors, returned for at least 1 recall examination, and whose parents gave consent for them to participate in the study. Color photographs were used for evaluation by 2 independent pediatric dentists. Parental satisfaction regarding the esthetics of the crowns was evaluated by a questionnaire. RESULTS: One hundred and twelve restorations placed in 40 children were evaluated. The evaluations were performed after the crowns had been in place for an average of 18 months (range=6-25 months). Overall parental satisfaction with the treatment was excellent; however, satisfaction with regard to color received the lowest rating. No significant differences were found between dentist and parent evaluations of color, size, and overall appearance (Fisher exact test; P=.194,.776,.291, respectively). Parents rated their overall satisfaction as being positive regardless of their poor ratings of color, size, or overall appearance. However, a significant relationship was found between durability and overall satisfaction (P=.046). Parents who gave poor ratings to durability also rated their overall satisfaction as being poor. CONCLUSIONS: Parental satisfaction with bonded resin composite SCs for the treatment of primary incisors with large or multi-surface caries was excellent. Parents' dissatisfaction was most often related to color of the restorations. However, this did not affect their overall satisfaction with the crowns. The durability of restorations negatively affected the rating of overall satisfaction with the crown. Durability seems to be of more concern than excellent color match to this group of parents. PMID- 15344628 TI - Orthodontic-surgical management of an impacted dilacerated maxillary central incisor: a clinical case report. AB - Dilaceration is one of the causes of maxillary central incisor eruption failure. Surgical excision is frequently the first choice of treatment for a severely dilacerated incisor. In this article, the case of a horizontally impacted and dilacerated maxillary central incisor was diagnosed and treated by surgical exposure using the apically repositioned flap technique combined with orthodontic traction. The dilacerated incisor was successfully moved into alignment, with pulpal vitality and periodontal health present 2 years following treatment. PMID- 15344629 TI - Localized aggressive periodontitis in a six-year-old: a case report. AB - The purpose of this report was to describe an approach to diagnose and effectively treat a pediatric patient with localized aggressive periodontitis. A 6-year-old female presented with clinical and radiographic evidence of severe attachment loss around several primary teeth. She had no history of systemic disease, periodontal disease, or caries prior to the periodontal abscess that prompted her referral. Routine immunological tests did not reveal any functional defects, but DNA testing for periodontal pathogens revealed the presence of all 8 aggressive periodontal pathogens assayed. Treatment consisted of the extraction of 2 severely affected primary teeth, increased frequency of recall appointments, and administration of systemic antibiotics. The patient's periodontal condition was stabilized 18 months post-treatment, and the 8 pathogens were no longer at detectable levels. With a treatment goal of preventing disease progression into the erupting permanent dentition, this treatment regimen provides an effective alternative to more aggressive strategies. PMID- 15344630 TI - Dentoalveolar trauma in a patient with chronic idiopathic thrombocytopenic purpura: a case report. AB - A case is presented of a 13-year-old boy with chronic idiopathic thrombocytopenic purpura (ITP) who sustained traumatic labial luxation of both lower central incisors, with partial alveolar fracture resulting in displacement of the labial alveolar plate. Intravenous immunoglobulin (Fleibogamma, 1 g/kg body weight x 2 days) was administered, resulting in the patient's platelet count rising from 15,000/mm3 to 70,000/mm3. Under general anesthesia, the displaced lower labial alveolus and luxated teeth were repositioned and splinted 2 days following trauma. Healing was uneventful. Subsequently, both lower central incisors became nonvital and were endodontically treated. The dental treatment of this patient with ITP is discussed in terms of emergency management, and subsequent care. PMID- 15344631 TI - Oral ulcerations as a sign of Crohn's disease in a pediatric patient: a case report. AB - Crohn's disease is an inflammatory intestinal disease of unknown etiology. The disease primarily affects whites, with both sexes being affected equally. A genetic predisposition exists. Symptoms frequently present in the second to third decades of life, although they may present in the pediatric and/or geriatric populations. Oral lesions are significant as they are frequently reported to precede intestinal symptoms. Treatment of Crohn's disease is palliative, with a focus on remission. Pediatric and general dentists play a critical role in the early diagnosis of Crohn's disease. Evaluation of a pediatric patient with complaints of oral ulcerations, as well as gastrointestinal symptoms, fatigue, and/or weight loss requires prompt referral to a gastroenterologist for further evaluation for Crohn's disease. PMID- 15344632 TI - Allergic reaction to intranasal midazolam HCl: a case report. AB - An acute allergic reaction in a 5-year-old healthy male, after receiving midazolam by intranasal atomizer for sedation purposes in the dental clinic, was reported. Shortly after the midazolam was provided, the child developed urticaria in both ankles, which rapidly progressed to the lower extremities, stomach, back, arms, neck, and face. The periorbital skin also became edematous. In the emergency room, the diagnosis of an urticaria allergic reaction was confirmed. The child was treated with intramuscular diphenylhydramine, discharged from the emergency room after 5 hours, and prescribed oral diphenylhydramine (Benadryl) and prednisolone (Orapred). Children who receive sedatives such as midazolam in the dental clinic should be carefully monitored from the moment they receive the sedative, in order to disclose and treat undesirable side effects of the sedative agents as early as possible. The implications of allergic reactions to sedative agents in the dental clinic are reviewed. PMID- 15344633 TI - Development of caries in permanent first molars adjacent to primary second molars with interproximal caries: four-year prospective radiographic study. AB - PURPOSE: This study investigated the effect of the primary second molars' distal surface caries on the incidence of the permanent first molars' mesial surface caries in 613 paired tooth surfaces of children ages 6 to 8 years at baseline examination. METHODS: Proximal caries and its progression were diagnosed by bite wing radiographs taken at a 1-year interval over a period of 4 years. The permanent first molars' mesial surfaces and primary second molars' distal surfaces were examined. Recorded were: (1) sound surfaces; (2) carious lesions on the enamel's external and internal half and on the dentin's external, middle, and internal third; (3) filled, extracted, and exfoliated teeth. The logistic model for panel data was employed to estimate the effect of proximal caries of the primary second molars' distal surfaces on the incidence of the permanent first molars' mesial surface caries. The 95% confidence interval probability was used. Sensitivity and specificity as well as the positive and negative predictive rates were computed. RESULTS: The results showed that the presence of proximal caries on each primary second molars' distal surfaces significantly affected the development of proximal caries on the corresponding permanent first molar's mesial surfaces. Age was estimated to exert a positive and highly significant impact, while gender had no effect. The odds ratio values ranged from 4.86 to 63.43. The values of sensitivity and specificity ranged from 45% to 97% and 80% to 89%, respectively, while the positive and negative rates ranged from 40% to 56% and 90% to 99%, respectively. CONCLUSIONS: Proximal caries present on the primary second molars' distal surfaces increases the risk of developing caries on the permanent first molars' mesial surfaces. This risk, however, is different among the paired surfaces studied. PMID- 15344634 TI - Self-reported awareness of unrestored dental caries. Survey of the Oral Health Status of Maryland Schoolchildren, 2000--2001. AB - PURPOSE: The purpose of this study was to assess and compare parental awareness of unrestored dental caries needs in a schoolchildren population according to socioeconomic status. METHODS: Subjective and normative unrestored dental caries data of kindergarten and third-grade children were collected from parental awareness questionnaires and clinical examinations, respectively, used in a 2000- 2001 statewide survey of Maryland schoolchildren. Descriptive, bivariate, and multivariate analyses were performed to assess awareness of unrestored dental caries. Parental perceived need also was compared to clinical findings of unrestored dental caries. RESULTS: Actual (clinically assessed) unrestored dental caries (21%) was found to be 2 times the level of self-reported awareness of dental caries (11%). Those most likely to have significantly incorrect awareness of unrestored dental caries were: (1) low-income populations; (2) residents of the Maryland Eastern Shore. Sensitivity was 34% and specificity 96% in comparing self-assessed, unrestored dental caries need with the clinical findings. CONCLUSIONS: An incorrect perceived oral health need is high in poorer populations and may serve as a barrier to their achieving optimal oral health, given that disease prevalence is higher and access to oral health care services is lower in this population group. PMID- 15344635 TI - Crossing the culture chasm: making the jump from medicine to management. AB - Recognizing the differences between the culture of business and the culture of clinical medicine is a key to success for business leaders. Learn how to identify and adapt to a new management culture. PMID- 15344636 TI - The written word: literature and leadership. AB - Explore what the current literature is saying about great leaders and leadership. Among the learnings: silence can hurt your organization, story telling is critical for inspiring employees and great leaders must be fearless. PMID- 15344637 TI - Want to be CEO? Focus on finesse. PMID- 15344638 TI - Resilient leadership for challenging times. AB - Examine six different leadership styes and consider how each one could be used in different circumstances to achieve favorable outcomes. Most physicians only use two of the styles and they may not always be appropriate. PMID- 15344639 TI - Inspired leadership. PMID- 15344640 TI - Test for better bedside manner fulfills Melnick's mission. PMID- 15344641 TI - Secrets of a chief medical officer. What they didn't teach you in medical school but you wish they had. PMID- 15344642 TI - Leadership insights of Xenophon. AB - Over two centuries ago, the Greek military historian Xenophon wrote the first systematic book on leadership, Anabasis, or March Up Country. According to management guru Peter Drucker, it is still the best book on the subject. Physician executives can learn timeless lessons from Xenophon. PMID- 15344643 TI - Physician leaders require special skills in a corporate environment. PMID- 15344644 TI - The changing landscape of pharmaceutical medicine. PMID- 15344645 TI - Standardizing physician-purchase preferences: from myth to reality. PMID- 15344646 TI - Physician's freedom of choice: fact or fiction? PMID- 15344647 TI - These things do happen--two broken systems. PMID- 15344648 TI - Convenience care: a patient-centered mode duling. PMID- 15344649 TI - How should we measure patient satisfaction? PMID- 15344650 TI - Tobacco policy: addressing public enemy number one. PMID- 15344651 TI - [Bioeffector sphingolipids as stimulators of cell growth and survival]. AB - The effects of various bioactive sphingolipids (sphingosine 1-phosphate, sphingosine 1-phosphocholine, ceramide 1-phosphate, ceramide beta-glucoside and beta-lactoside, and gangliosides) on cell proliferation and apoptosis are reviewed. It is concluded that the balance between the bioeffector sphingolipids determines their overall effect on cell. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344652 TI - [Leu-enkephalin homogeneously labeled with tritium in studying the Selank inhibiting effect on the enkephalin-degrading enzymes of human plasma]. AB - A method of analysis of enkephalinase activity in blood plasma based on the application of Leu-enkephalin generally labeled with tritium at all its amino acid residues was developed. The method allows the simultaneous estimation of activity of several peptidases in microquantities of tissues. [G-3H]Leu enkephalin was prepared by the method of solid phase catalytic isotope exchange (120 Ci/mmol) and subjected to proteolysis by the treatment with blood plasma. The resulting radioactive metabolites were separated by HPLC in the presence of the mixture of unlabeled fragments of Leu-enkephalin as internal standards. It was shown that aminopeptidases, dipeptidylaminopeptidases, and dipeptidylcarboxypeptidases respond for approximately 80%, 2%, and 10% of the total enzymatic activity, respectively. The new pathway of degradation of Leu enkephalin by carboxypeptidase that provides for approximately 6% of the total enkephalin-degrading activity was discovered. Bestatin was shown to predominantly inhibit aminopeptidases and carboxypeptidases, whereas Selank is more specific for carboxypeptidases and dicarboxypeptidases. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344653 TI - [The binding of Semax, ACTH 4-10 heptapeptide, to plasma membranes of the rat forebrain basal nuclei and its biodegradation]. AB - The binding characteristics of the peptide Semax (Met-Glu-His-Phe-Pro-Gly-Pro) to plasma membranes of basal nuclei of the rat forebrain and the dynamics of its degradation during its incubation with these membranes were studied. Binding of the homogeneously labeled [G-3H]Semax was shown to be time-dependent, specific, and reversible. Specific binding of the heptapeptide depended on calcium ions and was characterized by the dissociation constant of the ligand-receptor complex Kd = 2.41 +/- 1.02 x 10(-9) M and by the concentration of binding sites Bmax = 33.5 +/- 7.9 x 10(-15) mol/mg of protein. A method of studying Semax biodegradation in the presence of plasma membranes of rat brain was developed. It is based on the use of the peptide homogeneously labeled with tritium and on an HPLC analysis with UV detection at 220 and 254 nm of the peptide fragments formed. The half life of Semax in the presence of the plasma membranes was demonstrated to be longer than 1 h. Dipeptidylaminopeptidases are considered to be the main enzymes responsible for its biodegradation; they successively cleave Semax to the HFPGP pentapeptide and the PGP tripeptide. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344654 TI - [The release of flavin adenine dinucleotide upon local conformational transition in electron-transferring flavoprotein induced by trimethylamine dehydrogenase]. AB - The electron-transferring proteins, trimethylamine dehydrogenase (TMAD) and electron-transferring flavoprotein (ETF) from the bacterium Methylophilius methylotrophus, were studied in vitro by fluorescence spectroscopy. Flavin adenine dinucleotide (FAD) was found to be capable of a slow and spontaneous release from ETF, which is accompanied by an increase in flavin fluorescence. At a rather high ionic strength (0.1 M NaCl or 50 mM phosphate), the FAD release is sharply activated by TMAD preparations that induce a local conformational transition in ETF. The values of tryptophan fluorescence polarization and lifetime and the use of the Levshin-Perrin equation helped show that the size of protein particles remain unchanged upon the TMAD and ETF mixing; i.e., these proteins themselves do not form a stable complex with each other. The protein mixture did not release flavin from ETF in the presence of trimethylamine and formaldehyde. In this case, a stable complex between the proteins appeared to be formed under the action of formaldehyde. Upon a short-term incubation of ETF with ferricyanide, FAD was hydrolyzed to flavin mononucleotide (FMN) and AMP. This fact explains the previous detection of AMP in ETF preparations by some researches. A fluorescence method was proposed for distinguishing FAD from FMN in solution using ethylene glycol. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344655 TI - [Monomers for oligonucleotide synthesis with linkers carrying reactive residues: I. Synthesis of deoxynucleoside derivatives with methoxyoxalamide groups in heterocyclic bases]. AB - A number of monomers for the standard phosphoamidite oligodeoxynucleotide synthesis that carry reactive methoxyoxalamide groups attached to the thymidine, 2'-deoxycytidine, and 2'-deoxyadenosine heterocyclic bases were prepared. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344656 TI - [Monomers for oligonucleotide synthesis with linkers carrying reactive residues: II. Synthesis of phosphoamidites based of uridine and cytosine and containing a linker with methoxyoxalamide groups in position 2']. AB - A convenient preparative synthesis of 2'-amino-2'-deoxyuridine was developed. Starting from 2'-amino-2'-deoxyuridine and 2'-amino-2'-deoxycytidine, monomers for the phosphoamidite oligonucleotide synthesis were obtained that carry a linker with methoxyoxalamide groups in position 2'. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344657 TI - [New phosphonoformic acid derivatives of 3'-azido-3'-deoxythymidine]. AB - New 5'-alkyl ethoxy- and aminocarbonylphosphonates of 3'-azido-3'-deoxythymidine (AZT) were synthesized, and their antiviral properties in HIV-1-infected cell cultures and stability to chemical hydrolysis were studied. The AZT 5' aminocarbonylphosphonates were shown to be significantly more stable in phosphate buffer (pH 7.2) than the corresponding ethoxycarbonylphosphonates. The therapeutic (selectivity) index of some of the compounds exceeded that of the parent AZT due to their higher antiviral activity. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344658 TI - [Modification of the target cell surface with lipophilic glycoconjugates and interaction of the modified cells with natural killer cells]. AB - An experimental model system involving the modification of carbohydrate composition of the target cell surface with neoglycolipids was developed for studying the role of surface carbohydrates of target cells in the NK-cell mediated cytotoxicity. The polymeric glycoconjugates of the Glyc-PAA-PEA and Glyc PAA(Flu)-PEA types (where Glyc was an oligosaccharide residue, PAA poly(acrylamide) polymer, and PEA the phosphatidylethanolamine residue, and Flu fluorescein residue) capable of incorporation into the cell membrane were synthesized. The optimum structures of neoglycoconjugates and conditions for their incorporation into K562 and Raji cell lines, which differ in their sensitivity to the NK-cell-mediated lysis were selected. The mechanism of association of glycoconjugates with the plasma cell membrane and the kinetics of their elimination from the cell surface were investigated using the fluorescent labeled Glyc-PAA(Flu)-PEA derivatives. The spatial accessibility of the carbohydrate ligands for the interaction with human NK cells was demonstrated. The target cells modified with the Le(x) trisaccharide were shown to be more sensitive to the cytotoxic effect of human NK cells than the intact cells. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344659 TI - [Formylation reaction in series of meso-tetraaryl substituted porphyrins and chlorins]. AB - The formylation of nickel complexes of meso-tetraaryl substituted porphyrins and chlorins was studied. An obvious regioselectivity was observed for the introduction of the first and subsequent formyl groups into chlorins. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344660 TI - [Structural features of pregna-D'-pentaranes determining their interaction with three rat proteins]. AB - Competition of a number of progesterone 16alpha,17alpha-cycloalkane derivatives with 3H-labeled ligands for the binding sites of the rat uterine progesterone receptor, uterine pentaranophilin, and blood serum pentaranophilin was studied. We found that the selective ligands for the progesterone receptor are progesterone, 16alpha,17alpha-cyclopropanoprogesterone, and 16alpha,17alpha cyclopent-3'-enoprogesterone and the selective ligands for serum pentaranophilin are 6alpha-methyl-16alpha,17alpha-cyclohexanopregna-1,4-diene-3,20-dione and 3beta-hydroxy-16alpha,17alpha-cyclohexanopregn-5-en-20-one. No selective ligands for the uterine pentaranophilin were found. The majority of substituents in rings A, B, and D' we studied decreased the affinity of ligands for all the three proteins. The substitution of the delta5-3beta-hydroxy grouping for the delta4-3 keto grouping exerted the strongest negative effect in the case of the progesterone receptor and the uterine pentaranophilin, whereas the introduction of the 3',4'-dimethyl grouping strongly inhibited the ligand affinity for the uterine pentaranophilin. The extent and even the direction of the effect of a substituent on the affinity of ligands for the proteins substantially depended on the presence of other substituents in the steroid molecules. We hypothesized that a certain similarity exists between three proteins studied in respect to the structures of their ligand-binding pockets. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344661 TI - [The synthesis and antiviral activity of glycyrrhizic acid conjugates with alpha D-glucosamine and some glycosylamines]. AB - Glycyrrhizic acid and its 30-methyl ester were conjugated with 2-amino-1,3,4,6 tetra-O-acetyl-2-deoxy-alpha-D-glucopyranose, 2,3,4,6-tetra-O-acetyl-beta-D glucopyranosyl amine, 2,3,4-tri-O-acetyl-apha-L-arabinopyranosyl amine, 2 acetamido-2-deoxy-beta-D-glucopyranosyl amine, and beta-D-galactopyranosyl amine using N,N'-dicyclohexylcarbodiimide and its mixtures with N-hydroxybenzotriazole. Structures of the conjugates were confirmed by IR, UV, 1H, and 13C NMR spectroscopy. The glycoconjugate with the residues of 2-acetamido-2-deoxy-beta-D glucopyranosyl amine in the carbohydrate part of its molecule exhibited antiviral activity (ID50 4 microg/ml) toward the herpes simplex type 1 virus (HSV-1) in the VERO cell culture. Two compounds demonstrated anti-HIV-1 activity (50-70% inhibition of p24) in a culture of MT-4 cells at concentrations of 0.5-20 microg/ml. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344662 TI - [The effect of tetrazole and its amino derivatives on the kinetics of peroxidase oxidation of chromogenic substrates]. AB - The peroxidase-catalyzed oxidation of 2,2'-azino-bis(3-ethylbenzthiazoline-6 sulfonic acid) (ABTS), o-phenylenediamine (PDA), and 3,3',5,5' tetramethylbenzidine (TMB) was found to be activated by tetrazole and 5 aminotetrazole (AT) and weakly inhibited by 1,5-diaminotetrazole. The activating action of tetrazole and AT on the PDA and TMB oxidation was clearly discompetitive and that on ABTS was non-competitive. The coefficients (degrees) of activation alpha were determined for three substrates and two activators; they depended on the substrate type and the buffer nature and increased along with the pH growth from 6.4 to 7.2. For AT and tetrazole, the maximal alpha values were 4140 and 800 M(-1), respectively, upon the PDA oxidation and 3570 and 540 M(-1), respectively, upon the TMB oxidation. Lower alpha values (145 and 58 M(-1) for tetrazole and AT, respectively) were characteristic of the peroxidase oxidation of ABTS. The activation of peroxidase oxidation of the substrates by tetrazole and AT at pH > or = 5.4 was explained by the nucleophilic nature of the activators interacting with the amino acid residues in the peroxidase active site according to the mechanism of acid-base catalysis. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344663 TI - [Synthesis of modified psoralen analogues]. AB - Substituted 3-(5-methyl-7-oxofuro[3,2-g]chromen-6-yl)propanoic acids analogous to psoralen were synthesized by linear annulation of a furan moiety to the coumarin system. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344664 TI - [Aromatic crown ethers as phase transfer catalysts in the synthesis of N acetylglucosamine beta-aryl glycosides]. AB - The crown ether-catalyzed glycosylation of phenol, 4-methoxyphenol, and 4 nitrophenol was studied under phase transfer conditions in solid-liquid system. The asymmetric dibenzocrown esters are superior to [3.3]dibenzo-1 8-crown-6 and 15-crown-5 in the catalysis of these reactions. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. PMID- 15344665 TI - Poliomyelitis eradication in Africa--update on importations. PMID- 15344666 TI - Hepatitis B vaccines. PMID- 15344667 TI - Cambodia protects 75% of children against parasites. PMID- 15344668 TI - Psychogenic disorders: the consultants' responsibility, or "rule out" doctors. PMID- 15344669 TI - Rembrandt and the rat-catchers. PMID- 15344670 TI - The osteoarthritis initiative: an overview. PMID- 15344671 TI - Magnetic resonance imaging of osteoarthritis. PMID- 15344672 TI - Glucosamine for osteoarthritis: part I, review of the clinical evidence. AB - Glucosamine is a popular nutritional supplement for OA. This supplement has shown moderate efficacy in meta-analysis and large industry-sponsored clinical trials. However, smaller independent studies have not shown significant benefit. It is difficult to compare these clinical trials due to heterogeneity in trial design, differences in glucosamine products, and differences in osteoarthritic populations being studied. The National Center for Complementary and Alternative Medicine and the National Institute of Arthritis and Musculoskeletal and Skin Disease (NIAMS/NCCAM) have funded a multicenter five arm placebo controlled study called The Glucosamine Arthritis Intervention Trial (GAIT). GAIT spans 24 weeks, enrolling 1588 subjects, at 13 centers comparing the efficacy of glucosamine sulfate, chondroitin sulfate, glucosamine with chondroitin, to placebo and compared to celecoxib for knee OA. This study may have final data in March 2005. PMID- 15344673 TI - Glucosamine for osteoarthritis: part II, biologic and metabolic controversies. AB - Glucosamine is a popular nutritional supplement, discussed in clinical offices, peer-reviewed literature, and the media. Despite its popularity, much information is still needed regarding its biological effect in-vivo and its effects on glucose metabolism. The biologic effects of other constituents such as sulfate, chondroitin sulfate, and other additives need to be investigated as well. PMID- 15344674 TI - Double inlet left ventricle with transposition of the great vessels. PMID- 15344675 TI - Health risks and access to care among Rhode Island adults, 2002. PMID- 15344676 TI - A review of controlled substances. AB - Physicians who understand the Controlled Substances Act and their regulations will have fewer complications and problems. The tracking/monitoring system helps protect physicians and their patients from problems with abuse and diversion. The understanding that pharmacists are utilizing their skills everyday in reviewing a patient's pharmaceutical history and aiding the patient will no doubt encourage increased interaction among the healthcare fields and increase the quality of treatment for the patient. PMID- 15344677 TI - Miriam Hospital morbidity and mortality conference, a case of transient heart failure. PMID- 15344678 TI - Caring for survivors of childhood sexual abuse in medical practice. PMID- 15344679 TI - Complex issue of surgical outcome and hospital volume for several high-risk procedures. PMID- 15344680 TI - The ceaseless wandering of words. PMID- 15344681 TI - [Natriuretic beta-peptides in heart failure]. PMID- 15344682 TI - [Metabolic therapy in the combined treatment of patients with coronary heart disease]. PMID- 15344683 TI - [NSAID-associated hepatopathy]. PMID- 15344684 TI - [Coronary flow vasodilator reserve in patients with type 2 diabetes mellitus associated arterial hypertension]. AB - The coronary flow vasodilator reserve (CFVR) in the proximal segment of the anterior descending coronary artery was studied in 50 patients with diabetes mellitus (DM), by Doppler study via transesophageal approach. Group 1 included 39 patients with DM concurrent with Stages 1-2 arterial hypertension (AH), of them 14 patients were documented as having coronary heart disease (CHD) in the presence of coronary atherosclerosis (Subgroup 1A) and CHD was excluded in the remaining 25 patients (Subgroup 1B). Group comprised 11 patients with normal blood pressure (BP). For comparison, 6 healthy individuals were examined. CFVR was calculated as a ratio of the peak diastolic coronary flow (CF) velocity during infusion of dipyridamole (0.56 mg/kg) to the baseline CF. CDVR was significantly decreased as compared with the controls (2.07 +/- 0.73 in Subgroup 1A, 2.15 +/- 0.67 in subgroup 1B, 1.78 +/- 0.33 in Group 2, and 3.68 +/- 0.26 in the controls), this decrease being due to low CF velocities in hyperemia in the majority of patients in Subgroup 1A and Group 2 and to higher baseline CF velocity in most patients from Subgroup 1B. In Group 1 patients, CFVR was not linear with age, the duration of the disease, BP and HbA1 levels, but it was related to the carotid distensibility coefficient (rho = 0.60, p = 0.004) and to the blood level of total cholesterol (rho = -0.43, p = 0.0107). In Group 2 patients, the least CF velocities in the presence of vasodilatation were detectable in older patients and in patients with hypercholesterolemia. An all the patients with left ventricular hypertrophy (LVH) had decreased CFVR whose values with the myocardial mass index above 130 g/m2 were significantly less than those in the absence of LVH. Thus, the limited reserve of coronary vasodilatation was detectable in patients with DM irrespective of BP levels and the status of epicardial arteries and it was most pronounced in LVH and hypercholesterolemia. The impaired elastic properties of peripheral arteries in the presence of cholesterolemia may be regarded as a marker of the low reserve of coronary vasodilatation in patients with DM concurrent with AH. PMID- 15344685 TI - [Dilated cardiomyopathy resulting from asymptomatic and subclinical myocarditis]. AB - The paper presents data on 10 patients with dilated cardiomyopathy (DCM) or chronic myocarditis with fatal outcomes in whom heart failure was preceded by abnormal ECG changes originally regarded as the manifestations of coronary heart disease or small myocardial infarction. The clinical course of the disease and morphological studies suggest that the ECG changes were associated with pre existing asymptomatic viral myocarditis. PMID- 15344686 TI - [The parameters of life quality and the body mass index in elderly patients with essential hypertension]. AB - The parameters of life quality were analyzed in 22 elderly hypertensive patients with varying body mass (BM) in relation to age and sex. Worse life quality was observed with excessive BM and obesity in males and particularly in females. PMID- 15344687 TI - [The cardiovascular system in liquidators of consequences of the Chernobyl atomic power station accident]. AB - Liquidators of consequences of the Chernobyl atomic power station accident were found to have cardiovascular regulatory disorders in the late period. A late pressure reaction of systolic blood pressure to exercise was caused by cardiac rhythm rigidity due to the sick sinus syndrome. High exercise tolerance and delayed ischemia were indicative of diminished total peripheral resistance due to the impaired responsiveness of alpha-adrenoblockers in the arterial vessels and of the development of neurocirculatory dystonia. Cardiovascular functional changes in the liquidators are not attended by the higher morbidity rates of hypertensive disease. PMID- 15344688 TI - [New potentialities in the treatment of idiopathic hyperaldosteronism]. AB - The study was undertaken to examine whether the drugs affecting the renin angiotensin system may be used in the combined treatment of patients with idiopathic hyperaldosteronism (IHA). The study covered 20 patients (mean age 46.6 +/- 7.3 years). The diagnosis in all the patients was verified by a complex of instrumental studies, including the determination of plasma aldosterone concentrations (PAC) and plasma renin activity (PRA), and topical diagnostic studies (ultrasonography, computed tomography (CT)/magnetic resonance imaging (MRI), and angiography). The patients were given spironolactone, dihydropyridine calcium blockers, and beta-adrenoblockers, then with the cross-over method, this combination was added by the angiotensin-I receptor blocker (ATRB) losartane or the angiotensin-converting enzyme inhibitor (ACEI) lisinopril for 4 weeks. 24 hour blood pressure (BP) monitoring was used to evaluate the efficiency of therapy. In the combined therapy losartane, 50-100 mg/day, reduced systolic BP (SBP) by 8.4%, diastolic BP (DBP) by 11.1% while lisinopril, 10-20 mg/day lowered SBP and DBP by 7.6 and 9.9%, respectively. Both drugs improved the daily BP profile. When losartane or lisinopril added to the combined treatment, a therapeutic response was achieved in 15 (79%) and 14 (73.7%) patients, respectively. ARB and ACEI exert an antihypertensive effect in patients with IHA and they may be used in the combined treatment of this disease. The use of the above drugs may reduce the dose of spironolactone to the mean therapeutic one. The pathophysiological rationale for the use of ATRB for IHA is that the sensitivity of aldosterone secretion to the stimulating effect of angiotensin II is preserved despite the fact that the PAC is higher and PRA is lower. PMID- 15344689 TI - [Role of endothelial dysfunction and coagulation disorders in the development of pulmonary fibrosis in patients with interstitial lung diseases]. AB - Thirty-two patients with different forms of interstitial lung diseases (ILD), such as idiopathic fibrosing alveolitis (IFA) (n = 17) and fibrosing alveolitis concurrent with diffuse connective tissue diseases (FA-DCTD), were examined. Clinical, echocardiographic, computed tomographic, coagulative, and immunological studies were performed. Enzyme immunoassay was used to determine the levels of a complex of thrombin and antithrombin III (TAT) and platelet factor IV (PF-IV). There were significant increases in the levels of PF-IV (4.36 +/- 0.25 mg/l) and TAT (10.87 +/- 3.8 mg/l) in patients with ILD as compared to the control (2.75 +/ 0.47 and 1.8 +/- 0.2 mg/l, respectively; p < 0.05). In patients with early FA DCTD with the predominance of the milk glass syndrome during high-resolution CT (HRCT), the level of PF-IV was greater than the normal levels (p < 0.05) and decreased with the progression of the disease and with the formation of the honeycomb lung. If there were HRCT signs of active inflammation, the level of TAT was higher than that in the control; this was also in the development of irreversible fibrous changes. PMID- 15344690 TI - [Myocardial inotropic function in patients with chronic obstructive lung diseases as evidenced by biventricular equilibrium ventriculography]. AB - At present chronic obstructive lung diseases (COLD) occupy a prominent place in the morbidity and mortality of the adult population, with regarding the more and more increasing influence of aggressive environmental factors (sulfur dioxide, nitrogen, ozone, black smog and smoking). Due to the fact that clinical symptoms develop in patients with COLD at its late stages when there are irreversible changes in not only the bronchopulmonary apparatus, but, to a greater extent, the cardiovascular system; the efficiency of performed therapy is rather little. This shows it necessary to diagnose myocardial contractile disorders in such patients as soon as possible as the routine accessible cardiac study techniques (ECG, EchoCG) are of low informative value due to significant emphysema. In this view, of particular value is the use of radionuclide studies, including biventricular equilibrium radionuclide ventriculography that provides adequate information on the status of both ventricles. PMID- 15344691 TI - [Osteonecroses in systemic lupus erythematosus and antiphospholipid syndrome]. AB - The purpose of the study was to analyze the incidence of osteonecroses (ON) in systemic lupus erythematosus (SLE) patients with and without antiphospholipid syndrome (APS), primary APS (PAPS) and to define a relationship of the development of ON to some risk factors for vascular diseases. The study included 369 patients, including 293 with SLE, 160 with secondary APS, 76 with PAPS. The patients aged 14 to 63 years (mean 31.9+/-10.9 years). The history of disease was 0.6 to 30 years (mean 9.1+/-7.5 years). Among them 32 (8.7%) patients with aseptic necroses of different bones were selected in accordance with the data of X-ray studies. ON was detected in 8.7 patients with SLE and APS. The X-ray signs corresponded to third-to-sixth-degree ON. The most common site of ON was the head of the femur, although another site of ON was observed and multiple ON was typical. Leukopenia, creatininemia, fibrinogen levels were associated with ON (p < 0.05 by the Mann-Whitney test). The activity of SLE was significantly associated with SLEDAI scale scores (p < 0.05 by the Wald-Wolfovitz test). Six patients with SLE and ON had high scores by this scale--more than 40 scores of the maximum 150 possible scores. APS was diagnosed in 87.5 of the patients with ON (28 of the 32 patients) and only in 61.7% of the patients without ON (in 208 of the 337 patients) (chi2 = 8.4; p = 0.004). The development of ON in the examinees was significantly associated with the presence of APS. The activity of SLE, particularly nephritis, arthritis, positive tests for phospholipid antibodies, the presence of arterial thromboses, thrombocytopenia at the height of disease, therapy with large doses of glucocorticoids. PMID- 15344692 TI - [Use of alflutop in the treatment of patients with osteoarthrosis]. AB - The efficiency of alflutop was studied in patients with osteoarthrosis (OA) of the knee or hip joints. The examinees (n = 24) received alflutop only intramuscularly (in coxarthrosis) or intramuscularly in combination with the intraarticular injection of the agent (in knee osteoarthrosis). Physical examinations were made in control periods: before and immediately, 3, 6, and 12 months after treatment. Arthrosonography was used as an objective method for controlling the efficacy of the drug. The studies have demonstrated that alflutop substantially relieves pain, improves the function of the diseased joints in patients with OA. Arthrosonography has shown that the drug exerts an antiinflammatory effect, retards the progression of a pathological process in the joint. Alflutop has been found to be more effective in knee OA than in hip OA, which is determined by that its combined use regimen may be used in knee OA. The necessity and high efficiency of repeated courses of alflutop therapy are shown. PMID- 15344693 TI - [Device laboratory and postmortem parallels in alcoholic hepatitis during combined therapy using thioctic (alpha-lipoic) acid]. AB - The absence of clinical, biochemical, and instrumental signs that characterize the morphological pattern of the liver presents an urgent problem in the diagnosis of alcoholic hepatitis (AH). Sixty-one patients (mean age 41.7 +/- 8.7 years) were examined to study relationships between the histomorphological parameters and laboratory and instrumental findings in patients with AH on combined treatment including alpha-lipoic acid (Berlition). Indirect assessment of the histological activity of the disease may use the parameters of the serum activity of aspartate aminotransferase and gamma-glutamine aminotransferase, the stages of hepatic fibrosis-ultrasound symptoms: the splenic index at the early stages of the disease, the diameter of the splenic vein in moderate and severe fibrosis. The supplementation of Berlition in the combined therapy for AH decreases the severity of fatty hepatocytic dystrophy, hepatic inflammatory and necrotic changes, no increase or a decrease in the stage of fibrotic changes in the liver tissue. PMID- 15344694 TI - [Leveling the hyperlipidemic effect of beta-adrenoblockers by means of antiatherogenic vegetarian diet]. AB - The purpose of the study was to examine the capacities of correction of impaired lipid metabolism in patients with CHD receiving selective beta-adrenoblockers (beta-AB) by using an antiatherogenic milk-and-vegetable diet. According to the type of antiatherogenic diet, 67 patients were divided into 2 groups: 1) 42 patients were on an antiatherogenic vegetarian diet (a vegetarian group--VG) and 2) 25 patients received routine mixed diet No. 10c (a control group--CG). At the same time all the patients received similar antianginal drug therapy including the selective beta-AB atenolol in a dose of 50 mg/day. The vegetarian diet without special hypolipidemic therapy had a marked normalizing effect on the serum lipid spectrum in patients with CHD. Thus, in VG, by the end of treatment, the level of total cholesterol significantly decreased by 16% while in the controls it increased by 13%. High-density lipoprotein cholesterol increased in VG and decreased in CG, therefore the atherogenicity coefficient considerably rose. These were true for triglycerides and very low-density lipoprotein cholesterol. These parameters significantly decreased in VG (by more than 30%) and increased in CG (by 16%). Among the clinical symptoms, a more pronounced decrease in blood pressure in the patients on vegetarian diet and a more significant increase in their exercise tolerance. Balanced antiatherogenic milk and-vegetable diet in patients with coronary heart disease prevents the hyperlipidemic effect caused by the selective beta-AB atenolol and it is an agent for preventing its negative effect on lipid metabolism. PMID- 15344695 TI - [Efficacy of nebulized budesonide in severe exacerbation of bronchial asthma]. AB - The study was undertaken to examine the clinical effectiveness of nebulizer inhalations of budesonide suspension during severe attacks of bronchial asthma (BA) and the possibility of its use versus systemic glucocorticosteroids (GCSs). Sixty-eight patients admitted for a severe attack of BA were examined. The forced expiratory volume per sec (FEV1) was less than 40% and the peak forced expiration rate (PFER) was less than 150 l/min. First-line therapy included inhaled salbutamol (5-10 mg within 1.5-2 hours), oxygen, and intravenous prednisolone (120 mg). Thirty-two patients showed a good response to first-line therapy (a more than 50% increase in FEV1) as compared with the normal values) (Group 1) and 36 patients with concurrent obstructive lung disease (COLD) had an inadequate response (FEV1 was less than 40% of the normal values) was found in 32 patients (Group 2). Then the patients were randomized: 16 patients from Group 1 took budesonide, 4 mg/day, the other 16 received intravenous prednisolone, 120 mg/day. In Group 2, the efficacy of the drug added to therapy with oral prednisolone, 30 mg/day, was evaluated: 18 patients were given budesonide, 4 mg/day, and 18 received placebo. In patients with a good response to primary therapy, positive clinical and functional changes were found on day 6 of treatment. Group 2 patients receiving budesonide showed a significantly (p < 0.05) more marked positive changes in BA symptoms, FEV1, and PFER, which allowed a course of therapy with systemic GCSs to be reduced. Nebulized budesonide may be used instead of systemic GCSs to treat exacerbated BA if there is a good clinical and functional response to first-line broncholytic therapy, which is indicative of successful abatement of bronchospasm. If there is a poor response to first-line therapy in patients with more severe BA concurrent COLD, the early use of budesonide suspension in addition to systemic GCSs may enhance the efficiency of therapy for BA attacks and reduce the volume of systemic steroidal therapy. PMID- 15344696 TI - [Problems in the diagnosis of brucellosis in internal medicine]. PMID- 15344697 TI - [Advanced intestinal myiasis]. PMID- 15344698 TI - [A PET study of the brain mechanisms underlying perception of phrases with syntagmatic splitting]. PMID- 15344699 TI - [The association of EEG parameters with dominance of motivation of goal attainment during problem solving]. PMID- 15344700 TI - [Neuropsychological characteristics of cortical and subcortical zones of the brain as multidimensional genetic risk factors of schizophrenia]. PMID- 15344701 TI - [Gender differences in hemispheric spatiotemporal EEG patterns upon reproduction of verbal information]. PMID- 15344702 TI - [Electrophysiological mechanisms of the development of the adaptation processes]. PMID- 15344703 TI - [Linear blood velocity in arteries of the brain hemispheres in left-handers and right-handers during hypoxia]. PMID- 15344704 TI - [Changes in psychological and physiological parameters in young women during the individual annual cycle]. PMID- 15344705 TI - [Peculiarities of the heart rate regulation of the abiturients during entrance exams]. PMID- 15344706 TI - [Decoding of the transmembrane action potential into sodium, calcium, and potassium constituents]. PMID- 15344707 TI - [Changes in the heart rate parameters under the influence of academic school activities in adolescents with delayed psychological development]. PMID- 15344708 TI - [Evaluation of changes in correlation dimension of the dynamic series of RR intervals during a functional exercise test]. PMID- 15344709 TI - [Contractile cardiac activity in fetuses with intrauterine growth retardation: correlations between regional nonuniformity, relaxation, and afterload]. PMID- 15344710 TI - [Voluntary and involuntary control of breathing with imposed ventilation parameters]. PMID- 15344711 TI - [Regulation of external respiration and gas exchange during a 20-day exposure to sessions of intermittent normobaric hypoxia]. PMID- 15344712 TI - [Use of a mathematical model to analyze central hemodynamic parameters in humans under antiorthostatic conditions]. PMID- 15344713 TI - [The study of physicochemical parameters of the blood serum and renal function in children with some pathology of the internal organs]. PMID- 15344714 TI - [Relationships between platelets and blood cells in postprandial glycemia in healthy individuals]. PMID- 15344715 TI - [Influence of dosed traction on the mechanical-acoustic properties of the skin of human extremities]. PMID- 15344716 TI - [Age-related changes in turn-amplitude characteristics of the electromyogram recorded during graded isometric contraction]. PMID- 15344717 TI - [Protein adsorption on erythrocytic membranes and its effect on erythrocyte rheology in athletes during competition exercise]. PMID- 15344718 TI - [Evaluation of the excitability of motor neurons in the human cerebral cortex by magnetic stimulation method]. PMID- 15344719 TI - [Correlations between parameters characterizing the volume and number of erythrocytes and platelets in human blood]. PMID- 15344720 TI - [Relationship between hormones of the adrenal cortex, growth, and sexual development in schoolchildren]. PMID- 15344721 TI - [Mold fungi as disease agents. Sick people in sick houses?]. PMID- 15344722 TI - [Ingrown toenail makes walking painful. To wait, clip or cut?]. PMID- 15344723 TI - [Recurrent knee joint swelling. Not gout, although everything speaks in favor of it?]. PMID- 15344725 TI - [Smoking cessation. With hypnosis success in over 90%]. PMID- 15344724 TI - [Private medicine for insured patient. What do you think of individual health treatments?]. PMID- 15344726 TI - [Burns in children. First aid from the water tap? Better not!]. PMID- 15344727 TI - [Intersexual children. Who decides on their sex?]. PMID- 15344728 TI - [Type 1 and type 2 diabetes mellitus in Germany. An epidemic in all age groups]. PMID- 15344729 TI - [Newly diagnosed diabetes mellitus--what to look out for]. AB - Screening for diabetes makes good sense in particular in patients with overweight, hypertension or dyslipidemia. For type 2 diabetes is often not recognized until sequelae have put in an appearance. Consideration must be given to the possible presence of neuropathy, micro- and macroangiopathy and cardiovascular and cerebral disease. The primary therapy recommendations for type 2 diabetics comprise diet, weight loss and increased exercise. Depending on the success of these measures and the patient's constitution, medication with biguanides, sulfonylureas, glinides, glitazones alpha-glucosidase inhibitors or, where indicated, insulin, is then applied. PMID- 15344730 TI - [Practical diagnosis in insulin resistance and beta-cell dysfunction and resulting therapeutic consequences]. PMID- 15344731 TI - [Insulin pump treatment in children and adolescents with type 1 diabetes]. PMID- 15344732 TI - [Cell therapy and gene therapy in diabetes mellitus]. PMID- 15344733 TI - [Acute treatment of migraine in childhood]. PMID- 15344734 TI - [Flunarizine in the prophylactic treatment of childhood migraine]. PMID- 15344735 TI - [The risk of contracting hepatitis A or hepatitis B run by visitors to the Mediterranean and Eastern Europe]. AB - Hepatitis A and B continue to represent a risk for travelers. Those traveling to tropical countries are usually aware of this risk, while those visiting a Mediterranean country often are not. This investigation presents a destination related risk assessment and vaccination recommendations for travellers, on the basis of incidences and prevalences of hepatitis A and hepatitis B. For hepatitis B, the WHO classification based on HBsAg prevalence has been selected. For hepatitis A, countries have been categorized as low-, moderate- or high-risk. A comparison of infection risk with the numbers of airline passengers showed that in particular the large number of travelers to Tunisia, Turkey and Egypt, in conjunction with hygienic conditions in those countries, represent a source of imported hepatitis A. With regard to destinations in the Mediterranean and Eastern Europe, too, the risk of contracting hepatitis A or hepatitis B is not always negligible, so that vaccination may need to be recommended. PMID- 15344736 TI - [Sale of practice becomes more difficult. Therefore you raise the practice value]. PMID- 15344737 TI - [New specialist physician in internal and general medicine. You need testing, for EU doctors it falls into their laps]. PMID- 15344738 TI - [Everything hurts--fibromyalgia]. PMID- 15344739 TI - [Is it a birthmark?]. PMID- 15344740 TI - [From alcohol abstinence to transplantation. Can the liver still be helped?]. PMID- 15344741 TI - [Critical analysis of the medicine system: too much technology, too little soul]. PMID- 15344742 TI - [Bureaucratic madness in DMP is cleared up]. PMID- 15344743 TI - [The brain and the immune system in fluctuation. Why stress makes us sick?]. PMID- 15344744 TI - [Interdisciplinary action against lymphedema. Without compression the best lymph drainage is worth nothing]. PMID- 15344745 TI - [Prevention by life style intervention and medication. To run away from type 2 diabetes]. PMID- 15344746 TI - [Pilocytic astrocytoma in childhood]. PMID- 15344747 TI - [Individualized treatment strategies in adults with supratentorial WHO grade II gliomas]. PMID- 15344748 TI - [How useful are microsurgical operations in the treatment concept for glioblastoma multiforme]. PMID- 15344749 TI - [Nutrition and dietary treatment of chronic inflammatory bowel disease]. AB - The relationships between nutrition and chronic inflammatory bowel disease (CIBD) range from a possible involvement in the etiopathogenesis of the disease to dietary intervention in the various disease stages. CIBDs are often associated with dietary depletion. Prevention and treatment of malnutrition are therefore obligatory elements of therapy. The following stepped dietary treatment strategy applies: dietary counseling, supplementary dietary drinks, feeding via a stomach tube, parenteral alimentation. In the case of uncomplicated CIBD, an adequate and balanced diet (light full diet) is the goal. In the event of malnutrition, during an acute attack, and when complications occur, the diet must be adapted accordingly. Total parenteral alimentation is, per se, more likely to be associated with complications, is more expensive that enteral alimentation, and is thus reserved for specific indications. PMID- 15344750 TI - [Infant vomiting--differential diagnosis and management]. PMID- 15344751 TI - [Thrombosis associated with the Port-a-Cath system]. PMID- 15344752 TI - [Physicians' maintenance starts to crumble]. PMID- 15344753 TI - [Hessen physicians have a special problem. Additional annuities of contract physicians soon no longer financeable]. PMID- 15344754 TI - [New research results give hope. Causal therapy of Crohn disease in sight]. PMID- 15344755 TI - [Colon carcinoma. Outlooks of prevention useful]. PMID- 15344756 TI - [Coagulation self-control after valve replacement. Stable anticoagulation permits lowering of the intensity]. PMID- 15344757 TI - [First fixed combination of sartan and high-dosage diuretic. A strong duo against high blood pressure]. PMID- 15344758 TI - [Bone metastases in breast carcinoma. Bisphosphonate protects from complications]. PMID- 15344759 TI - [Thyrostatic agents are not always indicated. Treatment errors in overactive thyroid gland]. PMID- 15344760 TI - [New medication combines osteo-anabolic and anti-resorptive effects. Double strategy against osteoporosis]. PMID- 15344761 TI - [Counseling services help existential fears and everyday worries. Cancer patients need more than medical care]. PMID- 15344762 TI - [Turbid serum]. PMID- 15344763 TI - [Functional dyspepsia]. PMID- 15344764 TI - [Franz von Paula Gruithuisen -- the 230th birthday.. A Bavarian universal scientist and pioneer of lithotripsy]. PMID- 15344765 TI - Editorial: towards realistic water reuse practice for the "developed" and "developing" worlds. PMID- 15344766 TI - Towards sustainability in water recycling. AB - Those like us who believe in and spread the gospel of planned wastewater reclamation and reuse usually emphasize that this is a step towards sustainability in water resource management, but this is something that is very seldom analyzed. This paper discusses, from a critical point of view, issues such as goals in water reuse and influence on water demands, ecological analysis of the cycle of the main pollutants, health aspects and treatment requirements, energy consumption and measurable environmental benefits, in order to provide a set of criteria to assess sustainability in water recycling projects and to decrease the impact of the cultural water cycle on the environment. PMID- 15344767 TI - Strategic options for sustainable water management at new developments: the application of a simulation model to explore potential water savings. AB - Research on appropriate technologies and infrastructures to support water reuse has progressed rapidly over recent decades and there are now a wide range of source--treatment--reuse options for planners to choose from. Although the economics of water reuse schemes favours application to new developments rather than retrofit projects, there are few studies which have sought to address strategic option selection issues for large developments. The potential advantages of using treatment and reuse systems in new developments require an understanding of the relationships between a wide variety of social, environmental, technological, and operational factors. The operational effectiveness and economic efficiency of specific technology choices will vary as a function of network configuration, wastewater characteristics, how different technologies respond to dynamic loading (variability of feed strength and flow) and potential spiking, as well as equipment reliability, climate and household behaviour. Using a commercially available software package, the study reports the design and implementation of a low resolution simulation tool to explore sustainable water management options for a live case study site in the south of England (a peri-urban development of 4,500 new homes) with particular reference to opportunities for rainwater harvesting, and water reuse. PMID- 15344768 TI - Salt reduction in municipal sewage allocated for reuse: the outcome of a new policy in Israel. AB - Israel is presently reusing about 70% of its sewage and the buildup of salts in soils has been detected in some areas of the country. Prevention of sewage salt enrichment is one of the most immediately available solutions. The discharge of brines to municipal sewers is now prohibited and they are discharged to the sea. Discharge of brines to the sea has increased from 580,000 m3 in 1999 to more than 700,000 m3 in 2002. These salts are thus definitively taken out of the reuse cycle, year after year. The concentration of salts in industrial effluents is presently limited by the national water law and a unique regulation limiting the Boron and Sodium content in detergents has been implemented. Israeli industry has undergone a radical change in recent years. Many factories and all the hospitals have adopted K or Ca for softening and neutralization, while others have shifted to reverse osmosis. The concentration of Cl and Na in the sewage reaching treatment plants is declining. The addition of Cl to sewage has dropped from 120 mg/l to 70 mg/l. The average Boron concentration in sewage has dropped from 0.6 mg/l to 0.3 mg/l and should reach 0.2 mg/l by 2008. PMID- 15344769 TI - QMRA (quantitative microbial risk assessment) and HACCP (hazard analysis and critical control points) for management of pathogens in wastewater and sewage sludge treatment and reuse. AB - Hazard Analysis and Critical Control Points (HACCP) was applied for identifying and controlling exposure to pathogenic microorganisms encountered during normal sludge and wastewater handling at a 12,500 m3/d treatment plant utilising tertiary wastewater treatment and mesophilic sludge digestion. The hazardous scenarios considered were human exposure during treatment, handling, soil application and crop consumption, and exposure via water at the wetland-area and recreational swimming. A quantitative microbial risk assessment (QMRA), including rotavirus, adenovirus, haemorrhagic E. coli, Salmonella, Giardia and Cryptosporidium, was performed in order to prioritise pathogen hazards for control purposes. Human exposures were treated as individual risks but also related to the endemic situation in the general population. The highest individual health risk from a single exposure was via aerosols for workers at the belt press for sludge dewatering (virus infection risk = 1). The largest impact on the community would arise if children ingested sludge at the unprotected storage site, although in the worst-case situation the largest number of infections would arise through vegetables fertilised with sludge and eaten raw (not allowed in Sweden). Acceptable risk for various hazardous scenarios, treatment and/or reuse strategies could be tested in the model. PMID- 15344770 TI - Guidelines for the safe use of wastewater in agriculture: revisiting WHO guidelines. AB - The use of wastewater in agriculture is occurring more frequently because of water scarcity and population growth. Often the poorest households rely on this resource for their livelihood and food security needs. However, there are negative health implications of this practice that need to be addressed. WHO developed Guidelines for the Safe Use of Wastewater in Agriculture in 1989. The Guidelines are currently being revised based on new data from epidemiological studies, quantitative microbial risk assessments and other relevant information. WHO guidelines must be practical and offer feasible risk management solutions that will minimize health threats and allow for the beneficial use of scarce resources. To achieve the greatest impact on health, guidelines should be implemented with other health measures such as: health education, hygiene promotion, provision of adequate drinking water and sanitation, and other health care measures. PMID- 15344771 TI - Setting up microbiological water reuse guidelines for the Mediterranean. AB - Water reuse is a widespread practice in most Mediterranean countries. Some countries have no wastewater treatment facilities and direct reuse of raw wastewater is occurring while others have a well-established national reuse policy. Water reuse microbiological standards, when existing, significantly differ from one country to another. Some countries have adopted regulations close to the California's Water Recycling Criteria whereas other countries have chosen criteria based on the World Health Organisation (WHO) guidelines. California standards are technologically based requirements aimed at eliminating the presence of pathogens. The WHO guidelines relied on epidemiological evidences though few were available. Their revision on the basis of new epidemiological investigations and quantitative microbiological risk assessment (QMRA) provided by Blumenthal et al., together with added QMRA data, helped proposing Mediterranean guidelines. Acceptable annual risks related to bathing and potable water drinking were taken as benchmarks. This proposal is designed to protect individuals against realistic maximum exposures and to provide minimum and affordable requirements which should constitute the basis of water reuse regulations in every country of the region. Inadequacies of the actual knowledge do not allow a definitive position regarding the guideline limits; other scientific and technical basis are still required. PMID- 15344772 TI - Water reuse for irrigation from waste water treatment plants with seasonal varied operation modes. AB - Irrigation periods are usually limited to vegetation periods. The quality requirements for treated wastewater for disposal and for reuse are different. The reuse of water for irrigation allows partly the reuse of the wastewater's nutrients (N and P). Outside the irrigation period the water must be treated for disposal, thus nutrient removal is often required in order to avoid detrimental effects on the receiving surface water body. Only wastewater treatment plants with different operation modes for different seasons can realise these requirements. The nitrification is the most sensitive biological process in the aerobic wastewater treatment process. At low water temperatures the nitrifying bacteria need several weeks to re-start full nitrification after periods without NH4-removal. Therefore it is necessary to develop options for waste water treatment plants which allow a fast re-start of the nitrification process. Based on theoretical considerations and computer simulations of the activated sludge treatment process, one possibility for implementing a wastewater treatment plant with different seasonal operation modes is evaluated. PMID- 15344773 TI - Semi-intensive treatment plants for wastewater reuse in irrigation. AB - Semi-intensive technologies are a middle term between intensive ones (e.g., activated sludge with a retention time of hours) and extensive ones (e.g., stabilization ponds with a retention time of several weeks). The most common semi intensive configuration used in Israel is made of anaerobic ponds followed by aerated lagoons. These small low-energy units remove about 75-80% of the BOD and are followed by wastewater reservoirs for storage and complementary treatment. The reduction in loading allows a flexible operation of the reservoirs for the removal of other pollutants, while providing storage capacity to cope with the changes in water demand for irrigation during the year. In schemes for wastewater reuse in irrigation, this lay-out has proved to be low-cost, low-energy, flexible, reliable and efficient. Variations of this basic configuration are the use of UASB reactors instead of anaerobic ponds, aerated lagoons in series or low rate trickling filters instead of aerated lagoons, constructed wetlands or rock filters for algae removal, etc. Semi-intensive technologies use less energy than intensive ones, and less land than extensive ones. They can remove as much BOD as intensive ones, and as much pathogens and refractory pollutants as extensive ones. They release no or very small amounts of sludge. PMID- 15344774 TI - Role of irrigation and wastewater reuse: comparison of subsurface irrigation and furrow irrigation. AB - Two different irrigation systems, subsurface drip irrigation and furrow irrigation, are tested to investigate the level of viral contamination and survival when tertiary effluent is used in arid and semi-arid regions. The effluent was injected with bacteriophages of PRD1 and MS2. A greater number of PRD1 and MS2 were recovered from the lettuce in the subsurface drip-irrigated plots as compared to those in the furrow-irrigated plots. Shallow drip tape installation and preferential water paths through cracks on the soil surface appeared to be the main causes of high viral contamination in subsurface drip irrigation plots, which led to the direct contact of the lettuce stems with the irrigation water which penetrated the soil surface. The water use efficiency of the subsurface drip irrigation system was higher than that of the furrow irrigation system. Thus, subsurface drip irrigation is an efficient irrigation method for vegetable crops in arid and semi-arid regions if viral contamination can be reduced. Deeper installation of drip tapes, frequent irrigations, and timely harvests based on cumulative heat units may further reduce health risks by ensuring viral die-off under various field conditions. PMID- 15344775 TI - Reclaimed municipal wastewater as source of water and nutrients for plant nurseries. AB - A three year experimental activity was carried out to evaluate the possibility of reusing Reclaimed Wastewater (RWW) in Pistoia's nursery area (Central Italy). The research was aimed at: 1. identifying the best tertiary treatment facilities in order to make the effluent in compliance with the current Italian legislation for RWW reuse, 2. evaluating the effects of RWW irrigation on some ornamental, 3. quantifying fertilizing value and environmental impacts of RWW irrigation. In this paper the results of the last year experimental activity are presented. Experimental results indicated that filtration and disinfection with peracetic acid plus UV were very effective in indicator bacteria removal; as a matter of fact, neither Escherichia coli nor Total Coliforms were detected in any samples of pilot plant effluent. Results of an agronomical experiment indicated the suitability of the tertiary effluent for the irrigation: at the end of the growing season a higher (or equal) total dry mass was detected in RWW irrigated plants than well-water (WW) irrigated ones. Finally, the tertiary effluent can be considered as an important source of fertilizer for container-grown plants. As a matter of fact RWW irrigation showed to be able to replace at least one of the traditional fertilization treatments. PMID- 15344776 TI - Utilization of treated swine wastewater for greenhouse tomato production. AB - An integrated system has been developed to recycle waste organics and treated wastewater from a swine farm to make value-added products and to protect the environment from potential contamination. The farm is a farrow-to-wean swine operation with approximately 4,000 sows. A high-strength wastewater (chemical oxygen demand, 18,000 mg/l; total Khejdal nitrogen, 1,600 mg/l; total phosphorus, 360 mg/l) is produced from the swine operation. An ambient-temperature anaerobic digester has been used to treat the swine wastewater and to produce biogas (from an average 475 m3/day in winter to 950 m3/day in summer). The biogas is combusted in an engine to produce electricity (around 900 kW-hr/day). The digester effluent that is rich in nutrients (N, P, and minerals) is then utilized for fertigation for greenhouse tomato production. A trickling nitrification biofilter has been developed to convert ammonium in the effluent into nitrate. The nitrified anaerobic effluent is used as both fertilizer and irrigation water for approximately 14,400 tomato plants in greenhouses. Experimental data indicate that the tomato greenhouses have used approximately 12 m3 of the effluent and 3.84 kg nitrogen per day. At the same time, the greenhouses have a daily yield of 520 kg (37 g/plant) of marketable fruit. PMID- 15344777 TI - Electro-Fenton as a feasible advanced treatment process to produce reclaimed water. AB - The feasibility of the electro-Fenton process to generate simultaneously both of the Fenton's reagent species (Fe2+/H2O2), was assessed as a potentially more economical alternative to the classical Fenton's reaction to produce reclaimed water. An air-saturated combined wastewater (mixture of municipal and laboratory effluents) was treated in discontinuous and continuous reactors at pH = 3.5. The discontinuous reactor was a 2 L electrochemical laboratory cell fitted with concentric graphite and iron electrodes. The continuous reactor tests used a pilot treatment system comprising the aforementioned electrochemical cell, two clarifiers and one sand filter. Several tests were carried out at different conditions of reaction time (0-60 min) and electrical current values (0.2-1.0 A) in the discontinuous reactor. The best operating conditions were 60 min and 1 A without filtration of effluents. At these conditions, in discontinuous and continuous reactors with filtration, the COD, turbidity and color removal were 65 74.8%, 77-92.3% and 80-100%, respectively. Fecal and total coliforms, Escherichia coli, Shigella and Salmonella sp. were not detected at the end of the pilot treatment system. Electrogeneration of the Fenton's reagent is also economical; its cost is one-fifth the cost reported for Advanced Primary Treatment. PMID- 15344778 TI - Removal of detergents by activated petroleum coke from a clarified wastewater treated for reuse. AB - The removal of detergents from clarified wastewaters by activated petroleum coke (CAPA) was assessed. These substances, owing to their foamy properties, constitute a problem for ammonia removal by the air stripping process that could be installed in a wastewater treatment train to produce reclaimed water. CAPA was evaluated as a more economical alternative than a commercial activated carbon. Experimental work was divided in three stages: 1) production and characterisation of materials; 2) pretreatment of raw wastewater through the Fenton's reagent or coagulation-flocculation process with Al2(SO4)3; and 3) adsorption and bio adsorption tests of clarified effluents. These tests were carried out in the laboratory in discontinuous and continuous reactors, the former by the "point-by point" technique, with and without a previous fixing of bacteria, and the latter by the Rapid Small Scale Column Test. Detergents content, color, COD and UV254nm were measured in raw and treated wastewaters. Results show that the best pretreatment for the adsorption process was coagulation-flocculation rather than Fenton's method. Oxidation by this process decreased the adsorptive properties of detergents. Biomass fixed on the CAPA particles significantly increased the UV254nm and COD removal efficiencies (20% and 170% respectively). The breakthrough curves showed that CAPA could attain the expected detergents removal efficiency (66%) for the alum effluent. PMID- 15344779 TI - Combined membrane bioreactor (MBR) and reverse osmosis (RO) system for thin-film transistor-liquid crystal display TFT-LCD, industrial wastewater recycling. AB - In TFT-LCD industry, water plays a variety of roles as a cleaning agent and reaction solvent. As good quality water is increasingly a scarce resource and wastewater treatment costs rises, the once-through use of industrial water is becoming uneconomical and environmentally unacceptable. Instead, recycling of TFT LCD industrial wastewater is become more attractive from both an economic and environmental perspective. This research is mainly to explore the capacity of TFT LCD industrial wastewater recycling by the process combined with membrane bioreactor and reverse osmosis processes. Over the whole experimental period, the MBR process achieved a satisfactory organic removal. The COD could be removed with an average of over 97.3%. For TOC and BOD5 items, the average removal efficiencies were 97.8 and 99.4% respectively. The stable effluent quality and satisfactory removal performance were ensured by the efficient interception performance of the UF membrane device incorporated with biological reactor. Moreover, the MBR effluent did not contain any suspended solids and the SDI value was under 3. After treatment of RO, excellent water quality of permeate were under 5 mg/l, 2.5 mg/l and 150 micros/cm for COD, TOC and conductivity respectively. The treated water can be recycled for the cooling tower make-up water or other purposes. PMID- 15344780 TI - Treatment techniques for the recycling of bottle washing water in the soft drinks industry. AB - The soft drink production is an important sector in the manufacturing industry of Mexico. Water is the main source in the production of soft drinks. Wastewater from bottle washing is almost 50% of the total wastewater generated by this industry. In order to reduce the consumption of water, the water of the last bottle rinse can be reused in to the bottle pre-rinse and pre-washing cycles. This work presents the characterization of the final bottle washing rinse discharge and the treatability study for the most appropriate treatment system for recycling. Average characteristics of the final bottle wash rinse were as follows: Turbidity 40.46 NTU, COD 47.7 mg/L, TSS 56 mg/L, TS 693.6 mg/L, electrical conductivity 1,194 microS/cm. The results of the treatability tests showed that the final rinse water can be used in the pre-rinse and pre-washing after removing the totality of the suspended solids, 80% of the COD and 75% of the dissolved solids. This can be done using the following treatment systems: filtration-adsorption-reverse osmosis, or filtration-adsorption-ion exchange. The installation of these treatment techniques in the soft drink industry would decrease bottle washing water consumption by 50%. PMID- 15344781 TI - Membrane bio-reactor for advanced textile wastewater treatment and reuse. AB - Textile wastewater contains slowly- or non-biodegradable organic substances whose removal or transformation calls for advanced tertiary treatments downstream Activated Sludge Treatment Plants (ASTP). This work is focused on the treatment of textile industry wastewater using Membrane Bio-reactor (MBR) technology. An experimental activity was carried out at the Baciacavallo Wastewater Treatment Plant (WWTP) (Prato, Italy) to verify the efficiency of a pilot-scale MBR for the treatment of municipal wastewater, in which textile industry wastewater predominates. In the Baciacavallo WWTP the biological section is followed by a coagulation-flocculation treatment and ozonation. During the 5 months experimental period, the pilot-scale MBR proved to be very effective for wastewater reclamation. On average, removal efficiency of the pilot plant (93% for COD, 96% for ammonium and 99% for total suspended solids) was higher than the WWTP ones. Color was removed as in the WWTP. Anionic surf actants removal of pilot plant and WWTP were very similar (92.5 and 93.3% respectively), while the non-ionic surfactants removal was higher in the pilot plant (99.2 vs. 97.1). In conclusion the MBR technology demonstrated to be effective for textile wastewater reclamation, leading both to an improvement of pollutants removal and to a draw plate simplification. PMID- 15344782 TI - Alternative treatment strategy for tannery water reuse and material recovery. AB - Most tanneries use conventional systems for treatment of the mixture of all production effluents. Such an approach makes it possible to meet environmental regulations, but because of the high cost of the treatment facilities, its implementation has been scarce, especially in developing countries. With the waste reduction-elimination concept in view, an alternative strategy for water management is proposed based on individual treatment of the effluents from different processing steps to obtain multiquality recycled water for various reuse purposes, materials recovery and complete reuse of treated water. The methodology includes a database generation of tanneries in Mexico, a mass balance and pollution index determination, formulation of water management scenarios and technical-economical evaluation. To replace the traditionally used sulfde oxidation, a sulfide recovering was proposed. Chromium, grease and protein recovery were considered too. The proposed alternative allows a 90% fresh water reduction, the recovery of more than 95% of chromium and sulfide, 90% of grease, 65% of protein and zero discharge of wastewater. Simultaneous implementation of various water saving methods using in-house wastewater treatment techniques for recovering of chemicals and sub-products reduces substantially the cost of water management. PMID- 15344783 TI - Decontamination of wastewater contaminated by polychlorinated biphenyls (PCBs). AB - Wastewater contaminated by PCBs obtained from three different sources was treated at both laboratory and pilot plant scale conditions by ultraviolet oxidation of organics at the presence of hydrogen peroxide after partial adsorption of impurities and PCBs on activated carbon and/or activated bentonite. The procedure was conducted both with and without a Fe(II) catalyst and considerable reduction of PCB concentration was achieved in both cases. In pilot plant scale experiments, activated carbon polishing step followed UV oxidation. The following three types of contaminated waste water were examined: a) aqueous extracts originated in the course of clean-up of contaminated soil by extraction with aqueous solvents. Concentrations of PCBs in extracts were between 1 microg/L to 3,000 microg/L; b) wastewater condensates originated in the process of thermal desorption of PCB from soils. Concentrations of PCBs in condensates were between 300 microg/L and 5,000 microg/L. c) underground water contaminated by PCBs extracted from the sites of old contamination. The content of PCBs was up to 50,000 ng/L. Biodegradation of PCBs with a mixture of indigenous soil bacteria (selected strains of Pseudomonas and Acitenotobacter) was also tested. It was carried out in a reactor with volume of 1.5 m3 by application of the bacteria in a slurry of bentonite with adsorbed PCBs. PMID- 15344784 TI - Low salt petroleum produced water reuse: a farming alternative outside the food chain. AB - Oil and gas production gives rise to water production depending on the state of maturation of the Field. This means large volumes of water available. Today, this water is partly re-injected into the reservoir. Totalfinaelf's sustainable contribution to preserve water resources is to propose an alternative utilization: the reuse of produced waters from petroleum activities, outside the food chain. The aim of the first part of this study is to demonstrate the feasibility of utilizing low-salt water (<20 g/l) cleaned of hydrocarbons, for agricultural or forest irrigation. Rudimentary technologies such as artificial wetlands were tested to remove hydrocarbon substances and preliminary tests were performed with cotton (Gossypium hirsutum) and hemp (Cannabis sativa). Among the species tested in real conditions (greenhouse), hemp was affected by saline water whereas the results obtained for cotton were representative of the average worldwide production. These results validate the reuse of low-salt produced water in climatic conditions with expected temperatures of up to 37 degrees C in summer and 25 degrees C in winter. Following these results, field pilots are planned. Further research is planned to focus, taking into account local needs and environmental and production constraints. PMID- 15344785 TI - Anaerobic/aerobic treatment of colorants present in textile effluents. AB - The operation of an anaerobic/aerobic process used to degrade the colorants present in textile wastewater is presented. The objective is to produce water that can be reused. Two particular cases were studied: the degradation of a synthetic wastewater containing the colorant disperse blue 79 (DB79) as a model compound and a real textile effluent containing reactive azo dyes. The biodegradation was achieved using a single tank operated as sequencing batch reactor. It was observed that the DB79 was biotransformed to amines in the anaerobic stage decolorizing the wastewater. The amines formed were subsequently mineralized in the aerobic phase. An increase of toxicity was observed in the anaerobic stage due to the amines formation, but the wastewater was detoxified after the aerobic treatment. Removal efficiencies of DB79 around 92% were observed after the treatment. Around 96% of the initial color of the real wastewater was effectively removed. It was observed that the biomass pre acclimatized to the degradation of DB79 was more effective for the color removal than a freshly inoculum used. PMID- 15344786 TI - Grey water characterisation and its impact on the selection and operation of technologies for urban reuse. AB - Characterisation of grey water reveals a source water that is similar in organic strength to a low-medium strength municipal sewage influent but with physical and biodegradability characteristics similar to a tertiary treated effluent. The characteristics of the water suggest biological processes are the most suitable unit processes for treating grey water. The highly variable nature of the source requires that selected technologies must be inherently robust in their operation. One potential area of concern is the high COD/BOD ratio and nutrient deficiency in terms of both macro and micro nutrients which grey water exhibits potentially retard the efficacy of biological processes. PMID- 15344787 TI - Microbiological water quality in a large in-building, water recycling facility. AB - The Thames Water recycling plant at the Millennium Dome, London, reclaimed three sources of water: greywater from the washbasins, rainwater from the Dome roof and groundwater from a borehole on site. These were pre-treated separately, and the mixed stream filtered using ultrafiltration and reverse osmosis membranes. Monitoring for indicator microorganisms was undertaken throughout the plant and in the reclaimed water distribution system, as well as ad-hoc monitoring for the presence of pathogens in the raw waters. Treatment to the level of ultrafiltration was more than adequate to produce a water quality meeting existing worldwide reclaimed water guidelines for toilet flushing. Owing to the excellent quality of the water leaving the plant, no significant microbiological growth was observed in the reclaimed water distribution system during the year. The raw greywater exhibited a higher faecal bacterial load than the rainwater and groundwater, as predicted from more human contact (i.e. hand washing). Environmental strains of Legionella were observed in the three raw greywater samples analysed for pathogens, as was Cryptosporidium, Giardia and faecal enterococci. The rainwater had relatively high levels of faecal bacteria, probably of avian origin. Giardia was detected in one rainwater sample confirming the potential for this water source to contain pathogens. PMID- 15344788 TI - Domestic wastewater reclamation by submerged membrane bioreactor with high concentration powdered activated carbon for stream restoration. AB - This study focuses on the practical application of high concentration powdered activated carbon coupled membrane bio-reactor to domestic wastewater reclamation. The study was conducted in three parts, such as analysis of secondary domestic wastewater effluent, design and operation parameter evaluation and reclaimed water quality estimation for stream restoration. The organic concentration was 25.2-80.2 mgCOD(Cr)/L for the effluent of three domestic wastewater treatment plants. Around 50-75% of the COD was low molecular substances less than 1,000 which were quite biodegradable. The sawdust PAC was estimated to be proper adsorbent for the organics in the secondary effluents. Its Freundlich constant, K value was 5.847 and 1/n, 0.36. Using a system consists of single reactor with high concentration PAC (80 g/L) and submerged hollow fiber MF membrane module with nominal pore size of 0.1 microm, design and operation parameters were obtained, such as HRT of the bioreactor (2.5 hr), PAC concentration (80 g/L), the initial flux (less than 0.5 m/day) and intermittent suction cycle (12 min. suction and 3 min. idling). Organic removal by the system was high enough to produce reclaimed water for urban stream restoration The effluent organic concentration was at the level of 2 mg/L in terms of TOC (around 5 mg/L as COD(Cr)). Substances with molecular weight cut off < 1,000 were removed mostly by adsorption and biodegradation. Those above 1,000 were rejected at PAC cake layer on the membrane and gradually degraded by microorganisms during extended contact. PMID- 15344789 TI - Particle size distribution as a useful tool for microbial detection. AB - Worldwide, raw or treated wastewater is used for irrigation. However, this practice implies that the microbial content must be controlled. Unfortunately, detection techniques for microorganisms are costly, time consuming, and require highly trained personnel. For these reasons, this study used particle size distribution to measure the microbial quality of wastewater through correlations between the number or volume of particles and the concentration of fecal coliforms, Salmonella spp. and helminth ova. Such correlations were obtained for both raw and chemically treated wastewater. The best fit was the one for helminth ova, which applies for both the influent and effluent and also for all the coagulants involved. This technique allows the on-line quantification of helminth ova at a cost of US$3 and it takes only 5 minutes, instead of the US$70 and 5 days for the standard technique. With respect to the coagulants applied, their behavior is different only for particles smaller than 8 microm, and thus this value is considered as the critical size for this particular treatment. The best coagulant was the aluminium polychloride. In addition, this work establishes the distribution of COD, TSS, nitrogen, and phosphorous for particles smaller and larger than 20 microm. PMID- 15344790 TI - Ozonation by-products issued from the destruction of microorganisms present in wastewaters treated for reuse. AB - This work demonstrates the reaction of ozone on the amino acids comprising the covering layer of resistant micro-organisms. A secondary aim was to check the byproducts generated when ozone was applied to synthetic samples (such as Vibrio cholerae NO 01 WFCC-449, Salmonella typhi ATTC-6539, faecal coliforms and Ascaris suum). The ozone was applied at a concentration of 18.4 mgO3/min at pH 3, for different lengths of time. In the case of bacteria, results showed that, at 8 minutes, the number was reduced to the level of the Official Mexican Standards set for treated water destined for irrigation purposes (1,000 MPN/100 mL). Excellent correlation coefficients (0.95 to 0.99) were obtained for microbial concentrations versus ozone contact time. Destruction times required for 100% removal of the initial bacteria population varied between 2 and 14 minutes, while Ascaris suum required 1 hour. When Gram-negative bacteria die due to the effects of ozone, cellular lysis and the liberation of endotoxins (biodegradable) were observed. The ozonation of amino acids in the shell of Ascaris suum eggs, leads to the formation of aldehydes, such as formaldehyde and acetaldehyde, in low concentrations (0.0003 and 0.0005 microg/mL respectively). These levels are not hazardous to human health. PMID- 15344791 TI - Mathematical model for describing reactions of residual chlorine with organic matter in reclaimed wastewater. AB - Among several applications of urban wastewater reuse, use of reclaimed wastewater to sustain stream flows has become attractive in the urban area. Since these rivers are used for recreational purposes and for restoring aquatic eco-system, the adequate control of residual chlorine is essential. Mathematical model for describing reactions between residual chlorine and organic matter in reclaimed wastewater has been developed. The model considers the effect of molecular weight distribution of organic matter on the reaction rate. Lab-scale experiments were performed to estimate reaction rates constants and to examine their temperature dependency. The experiments showed that 1) the smaller organic matter gave the larger reaction rate; 2) temperature effect on reaction rate was described by the Arrhenius formula; 3) decline of free chlorine had more temperature dependency than combined chlorine. The comparison of computed results with data from lab scale experiments confirmed the validity of the model. We used the one dimensional dispersion model with proposed reaction model and examined the seasonal variation of residual chlorine profile along the river sustained by reclaimed wastewater in Sapporo. Simulation showed that seasonal variation of nitrification performance in secondary treatment as well as change in temperature caused seasonal variation in residual chlorine profile along the river. PMID- 15344792 TI - Technical and sanitary aspects of wastewater disinfection by UV irradiation for landscape irrigation. AB - Water reuse for landscape irrigation requires the production of high quality virus-free effluents to minimize risk for human health. In order to establish the relevance of MS2 phages as an appropriate biodosimeter for UV design, a pilot plant study has been carried out with different types of wastewater effluents. The two pilot systems tested (low-pressure high output and medium-pressure UV units) were able to achieve 4 and 5 log MS2 reduction in tertiary filtered effluent at high calculated UV doses of 170 _ 10 and 300 mJ/cm2, respectively. UV disinfection was extremely efficient for MS2 inactivation in high quality effluents after reverse osmosis: detention times as low as one second and UV dose of 40 mJ/cm2 were sufficient to reach 5 log inactivation of MS2. UV irradiation also produced rapid inactivation of human pathogens such as poliovirus type 1 and indigenous enteroviruses at UV doses up to 3 times lower that those for MS2 disinfection. It was concluded that accurate UV unit design for a given type of wastewater could be ensured by pilot tests using laboratory-propagated MS2 as biodosimeter and collimated-beam tests as the calibration-check. PMID- 15344793 TI - Contribution of the colmation layer to the elimination of coliphages by slow sand filtration. AB - River bank or slow sand filtration is a major procedure for processing surface water to drinking water in central europe. In order to model the performance of river bank and slow sand filtration plants, we are studying the different mechanisms by which the elimination of pathogens is realized. An important question concerning the mode of action of slow sand filters and river bank filtration units is the role of the colmation layer or "schmutzdecke" on the elimination of human pathogens. The schmutzdecke is an organic layer which develops at the surface of the sand filter short after the onset of operation. We have inoculated a pilot plant for slow sand filtration with coliphages and determined their rate of breakthrough and their final elimination. In the first experiment, with a colmation layer still missing, the breakthrough of the coliphages in the 80 cm mighty sandy bed amounted to ca. 40 %. In contrast, less than 1 % of coliphages escaped from the filter as the same experiment was repeated two months later, when a substantial colmation layer had developed. Our preliminary conclusions are that the colmation layer is extremely efficient in eliminating of viruses. PMID- 15344794 TI - The role of endocrine disrupters in water recycling: risk or mania? AB - The widespread occurrence of endocrine disrupting chemicals (EDCs), such as steroid hormones, in secondary wastewater effluents has become a major concern in the water recycling practice. This paper investigates the risk of steroid hormone breakthrough during nanofiltration membrane filtration in water recycling applications. The results indicate a dynamic equilibrium between adsorption and desorption of steroid hormone with regard to the membrane. This equilibrium can be pH dependent and there is a possibility for release of steroid hormones at high pH during membrane cleaning procedures or erratic pH variations. Increase in water recovery can severely increase the hormone breakthrough concentration. The results also indicate a possibility of accumulation of steroid hormones in the NF membrane, followed by subsequent release. PMID- 15344795 TI - Investigation on the behaviour of selected pharmaceuticals in the groundwater after infiltration of treated wastewater. AB - In a rural arid area without suitable receiving water the treated wastewater of a low loaded municipal wastewater treatment plant with full nutrient removal and additional post treatment steps is infiltrated into the unsaturated soil for groundwater recharge. Groundwater probes at increasing distances from the infiltration point have been sampled every two months over a period of 14 months. Beside conventional parameters (nutrients, carbon, and bacterial counts) samples were analysed for pharmaceutically active substances. Depending on and corresponding to their chemical structure and their fate during wastewater treatment, the selected substances showed different behaviour in the saturated zone of the groundwater. The antiepileptic Carbamazepine behaves very conservative and only is removed negligible even after long flow times within the subsurface zone. For other substances like the tranquilizer Diazepam or the analgesics Diclofenac a partial elimination during the different steps of wastewater treatment can be observed. Further degradation could be observed during the subsequent subsurface passage. In correlation with flow time additional removal of these substances from the aqueous phase can be observed. The musk substances Galaxolide and Tonalide were removed to some extend but not as good as the previous mentioned compounds. PMID- 15344796 TI - Removal mechanisms of endocrine disrupting compounds (steroids) during soil aquifer treatment. AB - The objective of this study was to determine the primary removal mechanisms of endocrine disruptors such as steroidal hormones present in reclaimed water, specifically 17beta-estradiol, estriol, and testosterone, during groundwater recharge via soil aquifer treatment (SAT). Steroidal hormones were quantified using enzyme-linked immunosorbent assays. Bench-scale studies and laboratory scale soil column experiments were employed to determine what mechanisms (i.e., adsorption, biodegradation, photolytic degradation) dominate the removal of the three compounds of interest during SAT. Findings of these studies revealed that the dominating removal mechanism for the compounds of interest during SAT is adsorption to the porous media matrix and additional attenuation to below the detection limit occurred in the presence of bioactivity. This additional removal occurred regardless of dominating redox conditions (aerobic vs. anoxic) or the type of organic carbon matrix present (hydrophobic acids, hydrophilic carbon vs. colloidal carbon). PMID- 15344797 TI - Pharmaceutical rejection by membranes for wastewater reclamation and reuse. AB - Various membranes, which have different materials and nominal molecular weight cut-offs (MWCO), were compared in terms of rejection of ibuprofen and removal of effluent organic matter (EfOM) from membrane bioreactor (MBR), because pharmaceutical compounds contain a potential risk and EfOM is the precursor of carcinogenic disinfection by-products when reusing for drinking water source. To provide equivalent comparison with respect to hydrodynamic condition, mass transfer parameter, J0/k ratio, was used. A tight-UF membrane with a molecular weight cut off of 8,000 daltons exhibited 25 approximately 95% removal efficiencies of ibuprofen with a molecular weight of 206 with and without presence of EfOM(MBR). EfOM(MBR) caused the reduction of ibuprofen removal efficiency for UF membrane. Rejection of EfOM(MBR) by UF and NF membranes ranged 29 approximately 47% and 69 approximately 86%, respectively. UF membrane could successfully remove ibuprofen at lower J0/k ratio range (< or = 1) in organic free water but could not efficiently reject ibuprofen with a relatively hydrophilic EfOM(MBR) (SUVA < or = 3). PMID- 15344798 TI - Assessing the removal potential of soil-aquifer treatment systems for bulk organic matter. AB - The fate of effluent organic matter (EfOM) during groundwater recharge was investigated by studying the removal behavior of four bulk organic carbon fractions isolated from a secondary effluent: Hydrophilic organic matter (HPI), hydrophobic acids (HPO-A), colloidal organic matter (OM), and soluble microbial products (SMPs). Short-term removal of the bulk organic fractions during soil infiltration was simulated in biologically active soil columns. Results revealed that the four organic fractions showed a significantly different behavior with respect to biological removal. HPI and colloidal OM were prone to biological removal during initial soil infiltration (0-30 cm) and supported soil microbial biomass growth in the infiltrative surface. Additionally, colloidal OM was partly removed by physical adsorption or filtration. HPO-A and SMPs reacted recalcitrant towards biological degradation as indicated by low soil biomass activity responses. Adsorbability assessment of the biologically refractory portions of the fractions onto powered activated carbon (PAC) indicated that physical removal is not likely to play a significantly role in further diminishing recalcitrant HPO-A, HPI and SMPs during longer travel times in the subsurface. PMID- 15344799 TI - Modeling the fate and transport of organic and nitrogen species in soil aquifer treatment process. AB - Soil aquifer treatment (SAT) is a promising technique for wastewater reclamation and reuse. This treatment strategy takes advantage of physicochemical and biological processes in the subsurface. The model employed in this study is based on MODFLOW-SURFACT (HydroGeoLogic, Inc.), a three-dimensional model for variably saturated flow and reactive mass transport. The model accounts for reactions including the nitrification of ammonium, the denitrification of nitrate, and the oxidation of organic carbon. Concentration of dissolved oxygen and biomasses involved in aerobic and anaerobic biological reactions forms the basis for estimates of nonlinear reaction rates formulated using a multiple-Monod expression. Illustrative simulations were conducted in a two-dimensional cross sectional domain, with unsaturated and saturated zones. They examine the effects that site and operational conditions have on the performance of a SAT system. The parameters and conditions of concern included length of the wet/dry cycle, ground surface condition, and infiltration rate. From the simulations, we found that organic carbon was effectively removed in all cases. The availability of oxygen was a key factor in predicting the production and removal of nitrate. Overall, the model successfully described the fate and transport of the key constituents during the wet/dry operational periods in both unsaturated and saturated subsurface. PMID- 15344800 TI - Sorption behaviors of heavy metals in SAT (soil aquifer treatment) system. AB - Degradation or detoxification of pollutants by SAT system was generally focused on organic contaminants although the regulation of water reuse has provisions of heavy metals. This study is to evaluate the feasibility of SAT for metals such as Cd, Cr and Pb with the pilot scaled column experiment. The desorption possibility of sorbed metals was also examined in the condition of continuous water and even acidic water (pH 4.3) injection. Fractional factorial analysis is a tool frequently used to identify factors or variables that have an effect on a certain response. A two-level fractional factorial analysis was planned to study the effect of four factors on Pb sorption from the experiments; soil particle size, TOC in solution, Pb concentration in solution and flow rate. The main effects among the factors were obtained by ANOVA based MINITAB software. The effects of four factors were also converted into coefficients; those values may build an empirical model to predict the metal sorption of soils. PMID- 15344801 TI - Quality assessment of an aquifer recharged with wastewater for its potential use as drinking source: "El Mezquital Valley" case. AB - Mexico City, with a population of 18 million, has been sending its wastewater for more than 100 years to the Tula Valley where it is used to irrigate 90,000 ha. Due to the large wastewater volume (60 m3/s) sent through unlined channels, combined with the use of very high irrigation rates, artificial recharge of the local aquifer has been occurring. This recharge is estimated in more than 25 m3/s. As a consequence, the water table has raised and several springs have appeared in the last decades with flows between 100 to 600 L/s. These springs and several wells are the water sources in the region. An evaluation of the Tula Valley aquifer quality was performed to analyze the use of such water as source of drinking water for Mexico City. The work is divided into 5 individual projects: (a) drinking water quality in the Tula Valley; (b) water availability in the Tula Valley; (c) wastewater treatment due to its use for irrigation, (d) use of membrane processes to treat groundwater; and (d) biota developed in the new surface water reservoirs. Results show that it is feasible to use this reclaimed water as drinking source. PMID- 15344802 TI - Effect of irrigation water quality on organic matter, Cd and Cu mobility in soils of Central Mexico. AB - Untreated wastewater has been used for irrigation since 1912 at the Irrigation District 03, Central Mexico. Accumulated heavy metals are dominantly bound to the organic soil fraction. In a field study we evaluated the effect of wastewater irrigation on the quality of soil organic matter and the amount of water extractable Cu and Cd. In a column experiment we tested if water treatment affects the leaching of both metals and dissolved organic carbon (DOC) out of soils that have been irrigated for more than 90 years with untreated wastewater. The field study shows that long term irrigation increases the mineralizable carbon fraction and the DOC concentrations. The water extractable Cu and Cd concentrations also increase and correlate with DOC. In the column leachates the Cu concentrations also correlate with DOC, the Cd concentrations correlate with the sum of cations, chloride and DOC concentrations. Water treatment reduced Cd leaching, but it did have no substantial effect on Cu leaching. PMID- 15344803 TI - Quality assessment of reclaimed water for its possible use for crop irrigation and aquifer recharge in Ensenada, Baja California, Mexico. AB - The city of Ensenada, Baja California, has three wastewater treatment plants and is one of the few cities in Mexico that treats all the wastewater that it generates. The largest wastewater treatment plant, called El Naranjo, treats on average 316 liters per second and complies with even the most stringent Mexican standards although a stricter control has to be achieved in order to avoid environmental and health problems At the moment, only 2% of the treated wastewater is used for the irrigation of sports fields and public landscape. The reclaimed water could be reused for the irrigation of crops for non-human consumption or ornamental products and/or for aquifer recharge. For reuse practices, two facts must be considered: a) an important part of the valley's production is exported to the USA and b) 30% of the city of Ensenada's water supply is obtained from the Maneadero aquifer. There is currently no Mexican legislation to stipulate adequate standards for aquifer recharge and decisions should be based on legislation from other places. Therefore, at the moment there is still a lack of technical and scientific elements to be able to make the best decision about the reuse of the wastewater. PMID- 15344804 TI - The reuse of treated wastewater for agricultural purposes in Nicaragua; Central America. AB - The first subsurface flow wetland (SSFW) system for about 1,000 PE, was constructed in Nicaragua in 1996 to apply this technology in the form of an integral project, combining the treatment of domestic wastewater with its reuse for crop production in small and medium size communities. The SSFW-effluent meets all standards established in the national regulations for wastewater reuse in agriculture, except for faecal coliforms, existent at an average concentration of 7 x 10(4) MPN/100 ml. A conventional surface irrigation method was used to irrigate different crop species selected to establish their risk of contamination. To judge the potential health risk for consumers and farmers, samples of vegetables and fruits harvested in the dry seasons of the years 1997 to 2002, were analyzed for the presence of pathogenic microorganisms like faecal coliforms, salmonella and shigella. In addition, a yield comparison between crops irrigated with well water using chemical fertilizers, and crops irrigated with the effluent of the SSFW-system was made, to analyze the economical benefits of the wastewater reuse. PMID- 15344805 TI - California's Task Force evaluation of issues confronting water reuse. AB - Water reclamation and reuse have been occurring in California for over a century. The state is arid and semi-arid in large regions and must rely on a variety of water supplies to meet its water demands. Reclaimed water is a potential source to meet a significant portion of new demands expected by 2030. However, there are issues of public health and water quality regulation, cost, public acceptance, institutional and other barriers that must be addressed. The California State Legislature adopted legislation in 2001 to convene the Recycled Water Task Force to advise the state on the opportunities for using reclaimed water in a broad spectrum of applications and in identifying impediments and constraints to increasing the use of reclaimed water. The processes for conducting the Task Force are described to illustrate the actions that were intended to increase the credibility of the Task Force. The recommendations of the Task Force are summarized. PMID- 15344806 TI - Current status of water reuse systems in Korea. AB - In Korea, the current water resources will fall short by 2.6 billion tons to meet the 38 billion ton water demand in the year 2020. To overcome the future water shortage, it is desirable to minimize water consumption and to reuse treated wastewater. There are a total of 99 on-site water-recycling systems in the country. The potential capacity of the 99 systems is 429 thousands tons/day, which is 3.6% of the total service water. Compared to other industrialized countries, the number of the water recycling systems in Korea is extremely small. This is mainly due to the following reasons. First, in Korea, any building with more than 60,000 m2 of total floor space is required to install a water reuse system by law. However, only less than 0.5% of the total buildings have more than 10,000 m2. Therefore, the regulation is ineffective and merely nominal. Second, service water is supplied at low charge (0.20 US-dollar/m3 water). The inexpensive service water often discourages people to recycle treated wastewater. Third, people still think recycled water is not clean enough and can cause diseases. Therefore, they should be informed that a well-maintained recycling system does not fail to produce water with high quality. PMID- 15344807 TI - Evaluation of wastewater effluents for soil aquifer treatment in South Korea. AB - Soil batch and column experiments were performed to characterize the wastewater effluents from seven different wastewater treatment plants in the Jonnam province, South Korea, with the purpose of evaluating the effluents for possible application of a soil aquifer treatment (SAT) in Korea. Batch experiments were conducted to measure the biodegradable dissolved organic carbon (BDOC) while 1 m soil columns, for simulating SAT, were employed to further analyze dissolved organic carbon (DOC) removal. The soils were collected from a river bottom in Jonnam. The BDOC fractions and the residual DOC concentrations for the effluents ranged from 19.3 to 59.9% and from 1.0 to 7.5 mg/L, respectively, depending on the reaction time. Applying the tentative criteria based on the data obtained for the BDOC and residual DOC, three effluents, from Gwangju, Hwasoon, and Jangsung, were found to be the most suitable for SAT applications. It was also concluded that the site characteristics should be also considered with regard to the retention time when evaluating the feasibility of SAT application in a certain region. PMID- 15344808 TI - Chihuahua: a water reuse case in the desert. AB - Water supply for all kind of uses in Chihuahua is mainly groundwater. During the last decade this city has been damaged with a heavy hydrologic crisis because of a persistent drought. This came up with the overexploitation of groundwater aquifers; therefore a deficit between demand and offer was done. To minimize this problem the government authorities have started an integral plan of optimizing hydrologic resources which considers the treatment of wastewater and the use of reclaimed water. The secondary wastewater treatment facility of the city treats about 30,000 m3/d of a wastewater with high organic contents, and produces an effluent with low concentration of suspended solids, organic matter, fats, detergents, and metals. Reclaimed water is conveyed toward strategic sites for the irrigation of great green areas in sport clubs, educational institutions and industrial zones, besides of its utilization on some manufacturing processes, road service, and also over construction industry. The potential reuse of this water goes farther from those activities; the treatment of the secondary effluent until the required levels of the water-bearing recharge criteria are met for drinking water supply is considered as the next step to achieve through a suitable planning strategy for the best integral resource advantage. PMID- 15344809 TI - Water reuse in the Apatlaco River Basin (Mexico): a feasibility study. AB - The aim of this work is to determine the technical and economic feasibility of implementing different reclamation and reuse projects that improve the quality of the Apatlaco river basin located in the central part of Mexico. A special methodology based on a decision support system was developed. This methodology allows to decide if it is convenient or not to finance a reclamation or reuse project for the most common water uses in the basin. This methodology is based on the net present value criteria (NPV) of the effective cash flow during the useful life of the project. The results obtained reveal a technical and economical feasibility for industrial reuse in Jiutepec and for agricultural reuse in Zacatepec and Emiliano Zapata. On the other hand, sanitation projects are not feasible in all cases analyzed. Therefore, Mexican Regulation (Ley Federal de Derechos en Materia de Agua) as currently implemented, does not promote and support this kind of projects. PMID- 15344810 TI - [Agro-ecosystems biodiversity and plant protection]. PMID- 15344811 TI - Management of disease resistance diversity of cultivars of a species in single fields: controlling epidemics. AB - The use of a diversity of resistance genes limits the development of polycyclic epidemics caused by airborne pathogens and reduces the risk that resistance be overcome by virulent races. Diversity can be easily achieved by growing mixtures of cultivars with different resistance genes and homogeneous agronomic traits. The mechanisms by which disease is reduced in cultivar mixtures include the loss of inoculum due to the presence of resistant plants between susceptible ones and resistance induced by avirulent pathogens. The complementary effects of individual mixture components reacting to disease pressure and to abiotic stresses result in greater yield stability compared with pure stands. The quality of products from mixtures is at least equal to that obtained with pure stands. This type of resistance management is applicable to both annual and perennial crops. PMID- 15344812 TI - [Specific diversity of plant populations at rainfed scale and crop protection: the example of banana production in the French West Indies]. AB - Banana is a major crop in the French West Indies, where it is subjected to strong parasite pressure, resulting in pesticide pollution. An increase in plant population diversity in the cultivated ecosystem is generated by changing cultural practices. This results in a decrease in parasite pressure and hence a decrease in pollutant pesticide loads. Agricultural sustainability is therefore reinforced for better coexistence of populated, cultivated and protected areas. PMID- 15344813 TI - Plant parasite control and soil fauna diversity. AB - The use of pesticides to control plant parasites and diseases has generated serious problems of public health and environmental quality, leading to the promotion of alternative Integrated Pest Management strategies that tend to rely more on natural processes and the active participation of farmers as observers and experimenters in their own fields. We present three case studies that point at different options provided by locally available populations of soil organisms, the maintenance of diverse populations of pests or increased resistance of plants to pest attacks by their interactions with earthworms and other useful soil organisms. These examples demonstrate the diversity of options offered by the non planned agro-ecosystem diversity in pest control and the need to identify management options that maintain this biodiversity. PMID- 15344814 TI - Symbiotic microorganisms, a key for ecological success and protection of plants. AB - Plant-associated microbial diversity encompasses symbionts, protecting their host against various aggressions. Mycorrhizal and rhizospheric microorganisms buffer effects of soil toxic compounds and soil-borne pathogens. Endophytic bacteria and fungi, some of which are vertically inherited through seeds, take part in plant protection by acting directly on aggressive factors (mainly pathogens and herbivores) or by enhancing plant responses. Plant protective microbial symbionts determine the ecological success of plants; they drastically modify plant communities and related trophic webs. This review suggests approaches to improve the inventory of diversity and functions of in situ plant-associated microorganisms. PMID- 15344815 TI - Oxidative stress: a theoretical model or a biological reality? AB - Although oxidative stress has been extensively studied the last fifteen years, many physicians and biologists are still sceptical concerning its interest in biology and medicine. This is probably due, in part, to the fact that this subject is a matter of biophysics, and the first studies reported were written using a physical language that inspired these people used to a more concrete problematic very little. Another problem is the difficulty to detect the species mediating oxidative stress, and to determine their role in biological processes. This review is aimed at presenting oxidative stress, as well as reactive oxygen species and free radicals--the molecules that mediate it--in a clear form able to convince all researchers involved in life sciences that these short-lived intermediates are indissociable from any aerobic organism. Moreover, if reactive oxygen species and free radicals are undoubtedly involved in many pathologies, they have physiological functions too. PMID- 15344816 TI - About the equilibrium shape of fibred structures, and biological shapes. AB - Biological morphogenesis has often been modeled with reaction-diffusion models [A.M. Turing, The chemical basis of morphogenesis, Phil. Trans. R. Soc. Lond. B 237 (1952) 37-72]. The interplay of bio-chemical fields is supposed to generate shapes by positional information carried by the values in the field. However, the structure of the biological tissue at the microscopic scale is absent from these models. We show that the fibred nature of biological tissue induces specific morphogenic properties. Fibred shapes can be calculated from physical principles borrowed from the theory of crystallogenesis. These give an intuitive insight into the shape of fruits or vegetables, buds and pins in botany, fingers, muscles, insects abdomen and heart in the animal realm, and also into other fibred structures such as the mitotic spindle. We predict the existence of bumps, apices or cusps at poles of fibred structures. An extrapolation to out-of equilibrium growth predicts that these structures grow forward in the direction of the cusp, and that fibred organs should have a regular branching ordering. However, our model does not take into account the elasto-plastic properties, or the composite nature of the living material. PMID- 15344817 TI - Iron-superoxide dismutase and monodehydroascorbate reductase transcripts accumulate in response to internode rubbing in tomato. AB - A cDNA encoding an iron-superoxide dismutase (Fe-SOD) was isolated by RACE-PCR from a Lycopersicon esculentum cDNA library. The Fe-SOD cDNA consists of a 746-bp open reading frame and is predicted to encode a protein of 249 amino acids with a calculated molecular mass of 27.9 kDa. The deduced amino acid sequence was very similar to other plant Fe-SODs and a potential chloroplastic targeting was found. To study the induction of oxidative burst in response to mechanical stimulation, the accumulation of Fe-SOD and monodehydroascorbate reductase (MDHAR) mRNAs was analysed in response to young growing internode rubbing in tomato plants. Northern analyses show that Fe-SOD mRNA and MDHAR mRNA accumulated in tomato internodes 10 min after the mechanical stimulation. These results suggest that reactive oxygen species are early involved in the response of a plant to a mechanical stimulation, such as rubbing. The nucleotide sequence data reported in this paper will appear in the NCBI Nucleotide Sequence Databases under the accession number AY262025. PMID- 15344818 TI - Growth and cellular fatty-acid composition of a sulphate-reducing bacterium, Desulfatibacillum aliphaticivorans strain CV2803T, grown on n-alkenes. AB - The anaerobic degradation of n-alkenes by a sulphate-reducing bacterium Desulfatibacillum aliphaticivorans strain CV2803T was investigated. Results suggest that enzymes required for alkene degradation are inducible. Moreover, total cellular fatty acids of strain CV2803T were predominantly C-odd when the strain was grown on C-odd substrates and C-even when grown on C-even substrates. In addition to classical bacterial fatty acids, unusual 4-Me-17:1delta11 and 4-Me 18:1delta11 fatty acids and their saturated homologues were detected when strain CV2803T was grown on 1-pentadecene and 1-hexadecene, respectively. These methyl branched monounsaturated fatty acids could constitute specific metabolites of n alkene degradation by sulphate-reducing bacteria. PMID- 15344819 TI - [Subperitoneal cyst in a Friesian mare]. AB - The case of a 5-year-old Friesian mare with a mass in her abdomen and a gait irregularity of the left hind limb is discussed. At rectal ultrasonogrphic examination the mass appeared to be a thin-walled, anechogenic cavity in the left abdomen. By means of laparoscopy the mass could be visualized as a smooth structure, covered by peritoneum and fat. Under laparoscopic guidance the mass was punctured and fluid was aspirated. Cytological and bacteriological findings of the fluid were indicative of a cyst. After drainage of the cyst the locomotion disorder of the left hind limb disappeared. It is concluded that this horse suffered from a subperitoneal cyst, a rare disorder in humans and never previously described in horses. The case emphasizes the value of laparoscopy in the diagnosis and treatment of (intra)abdominal masses and further clearly shows that in the differential diagnosis of locomotion disorders not only structures related to the locomotion system should be taken into account. PMID- 15344820 TI - [Go Holland!]. PMID- 15344821 TI - [The General Food Law: the new European hygiene law]. PMID- 15344822 TI - [The job and the way of working of the veterinarians in mass vaccination: what is the decision of the court?]. PMID- 15344823 TI - [In fifteen years less interest in companion animal veterinarians? Interview with veterinarian Richard Rieter about the future of veterinary medicine in the Netherlands. Interview by Marianne M. Sloet]. PMID- 15344824 TI - [Legal cases, what do you mean?]. PMID- 15344825 TI - [Tall, healthy and happy veterinarian]. PMID- 15344826 TI - [Distribution of orphan drugs by the pharmacist of the Faculty of Veterinary Medicine]. PMID- 15344827 TI - [Book: The Rabbit from the series About Animals]. PMID- 15344828 TI - ["100 questions about animal diseases": a critical point of view]. PMID- 15344829 TI - [Castration of swine with anesthetics?]. PMID- 15344830 TI - [Always hitting?]. PMID- 15344831 TI - [General secretary gave 'taking in stride' a new meaning during his stay with KNMvD. Dr. Tjeerd Jorna: "Veterinarians cannot exist without one another"]. PMID- 15344832 TI - [Farewell word from Tjeerd Jorna]. PMID- 15344833 TI - [Oxygen for the Dutch veterinarian]. PMID- 15344834 TI - [Job inspection after implementation of the X-ray legislation]. PMID- 15344835 TI - [Review of availability and demand in employment vacancies for January-April]. PMID- 15344836 TI - How doctors help the dopers. PMID- 15344837 TI - The flu connection. Does prenatal exposure to influenza raise the risk of schizophrenia. PMID- 15344838 TI - Synthesis of some mono-Mannich bases and corresponding azine derivatives and evaluation of their anticonvulsant activity. AB - Mono-Mannich bases, 3-amino-1-aryl-1-propanone hydrochlorides (Ig1-Ig4), and their corresponding azine derivatives, N,N'-bis(3- amino-1-aryl-propylidene) hydrazine dihydrochlorides (D1-D4), were synthesized and their anticonvulsant activities were evaluated. Alterations in biological activity depending on modifications in chemical structure were also followed. The aryl part was phenyl in Ig1, D1, Ig2, D2, Ig3, D3, or p-hydroxyphenyl in Ig4, and D4. The amine part was dimethylamine in Ig1, D1, Ig4, and D4, piperidine in Ig2, D2 or morpholine in Ig3, D3. Compounds D2, D3, and D4 are new. The anticonvulsant activity was determined by maximal electroshock (MES) and subcutaneous metrazole (pentetrazol; scMet) tests. The rotorod toxicity test was used for determining neurological deficits. While the compounds were not effective in scMet, they were found to exert protective effect in MES. The results of MES are as follows: Compound [dose level (mg/kg), time (h)]: Ig1 [30 (0.5 h), 100 (0.5 h)]; Ig2 [30 (0.5 h, 4 h)]; Ig3 [30 (0.5 h), 100 (0.5 h), 300 (0.5 h, 4 h)]; Ig4 [300 (0.5 h, 4 h), 100 (4 h)]; D1 [30 (0.5 h)]; D3 [100 (0.5 h,4 h), 300 (0.5 h), 30 (4 h)]]; D4 [300 (0.5 h, 4 h)]. D2 did not show any anticonvulsant activity in both tests. Ig1, Ig2, D1, D2, and D3 exhibited neurotoxicity. Compounds Ig2, D1, and D2 were neurotoxic at 100 mg/kg dose level at 0.5 h. Ig1 was neurotoxic at 300 mg at 0.5 h, D3 was neurotoxic at 300 mg at 4 h. Conversion of mono-Mannich bases to their corresponding azine derivatives generally decreased the anticonvulsant activity. Ig3, Ig4 and D4 seem to be promising candidates to develop new anticonvulsant compounds for grand mal epilepsy for further synthesis and in vivo studies, since they were effective in MES screening and no neurotoxicity was observed with them. PMID- 15344839 TI - Behavioural effects of thieno and pyrazolo [2,1] benzothiazepine derivatives in mice. AB - Behavioural studies were conducted in mice with a number of hetero[2,1]benzothiazepine derivatives, analogues of tianeptine. Previously published studies in mice have shown that some of these compounds were effective in the tetrabenazine and Porsolt tests. In the present study, four of the 15 compounds under study potentiated the actions of 5-hydroxytryptophan (5-HTP, 50 mg/kg i.p.), but no significant antagonism of the apomorphine (16 mg/kg s.c.) induced hypothermia and potentiation of the amphetamine actions was found. Moreover, some of them inhibited the stereotyped behaviour and/or climbing behaviour of low doses of apomorphine and compound 2 was effective in the plus maze test. These compounds also produced a slight inhibition of exploratory behaviour in the holeboard test. On the other hand, no significant muscle relaxant and anticonvulsant activities were observed at any dose employed. Together, these data suggest that some of the compounds under study combine the antidepressant effects with additional neuroleptic or anxiolytic activities in mice. PMID- 15344840 TI - Studies on 3-diethylaminocarbonyl-1,4,5,6,7,8-hexahydroquinoline derivatives and their calcium channel antagonistic activities in vitro. AB - In this study, thirteen 2,6,6-trimethyl-3-carbamoyl-4-aryl-5-oxo-1,4,5,6,7,8 hexahydroquinoline derivatives were synthesized and screened for their calcium channel antagonistic actitivities. The hexahydroquinoline derivatives were synthesized according to Hantzsch reaction. The structures of the compounds were elucidated by IR, 1H-NMR, 13C-NMR, Distortionless Enhancement Polarization Transfer (DEPT), Correlation Spectroscopy (COSY), mass spectroscopy and elemental analyses. The calcium antagonistic activities of the compounds were determined by tests performed on isolated rat ileum and lamb carotid artery. In isolated rat ileum, compounds 8, 12 and 13 were found to be more active than nicardipine (CAS 55985-32-5) at a concentration of 10(-5) mol/L. At the concentration 10(-4) mol/L, compounds 7 and 12 were more active than nicardipine and compounds 5, 8 and 13 were equipotent. In lamb carotid artery studies, at the concentration 10( 5) mol/L all compounds were found to be less active than nicardipine; at the concentration 10(-4) mol/L compounds 5 and 13 showed greater inhibition than nicardipine. At this concentration, compound 9 was found to be as active as nicardipine. PMID- 15344841 TI - Determination of alpha-tocopherol in plasma by high performance liquid chromatography with fluorescence detection and stability of alpha-tocopherol under different conditions. AB - In order to study the clinical effects of the antioxidant alpha-tocopherol (tocofersolan, CAS 30999-06-5) a simple, robust, sensitive and reliable high performance liquid chromatography (HPLC) method using fluorescence detection for the daily measurement of alpha-tocopherol concentration in plasma is described. Several methods of HPLC analysis using fluorescence techniques are published, however, investigations of the stability of alpha-tocopherol during sample preparation under different conditions are relevant in the routine measurements and have to date not been reported. Therefore a modified method, both sensitive and robust, qualified for the day-by-day measurement of alpha-tocopherol in plasma was established and the stability of alpha-tocopherol during sample preparation was analysed under different conditions. This method enables the complete separation of alpha-tocopherol from other compounds within 14 min. In addition, investigations on the stability of alpha-tocopherol during sample preparation showed the necessity of rapid sample preparation without delay. Prepared samples can be stored in liquid nitrogen for at least 56 days. PMID- 15344842 TI - Metabolic stability and uptake by human hepatocytes of pitavastatin, a new inhibitor of HMG-CoA reductase. AB - To gain a better understanding of the metabolic stability and transport of pitavastatin (CAS 147526-32-7), a new and potent 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, experiments were conducted using human hepatocytes and oocytes of Xenopus laevis expressing human organic anion transporting polypeptide-2 (OATP2), respectively. Almost the entire radioactivity was from the unchanged substance or lactone form in human hepatocytes, and the cytochrome P450 (CYP)-mediated metabolism of pitavastatin was negligible. The results suggested that CYPs are not critically involved in determining the metabolic fate of pitavastatin. The hepatic uptake of pitavastatin reached saturation with a Km of 2.99 +/- 0.79 micromol/L. Also, the uptake of pitavastatin was mediated by OATP2 expressed in oocytes with a Km of 5.53 +/- 1.70 micromol/L. These results indicated that OATP2 plays a major role in the distribution of pitavastatin in liver. Furthermore, to elucidate the increase in the plasma concentration of pitavastatin in a clinical setting, the inhibitory effect of ciclosporin (cyclosporin A, CAS 59865-13-3) on the uptake of pitavastatin was examined. The uptake of pitavastatin was inhibited in the presence of cyclosporin A and the apparent IC50 value was 2.91 +/- 0.78 micromol/L. This result may at least partly explain the drug-drug interaction between pitavastatin and cyclosporin A. In conclusion, the characterization of transporters needs to be taken into account to avoid transporter-mediated drug drug interaction. PMID- 15344843 TI - Comparison of the inhibition of myeloperoxidase-catalyzed hypochlorite formation in vitro and in whole blood by different plant extracts contained in a phytopharmacon treating functional dyspepsia. AB - Ethanolic extracts from nine medicinal plants are combined in Iberogast (IG). This phytomedicine is successfully used in the treatment of gastrointestinal disorders. Functional gastrointestinal diseases such as non-ulcerous dyspepsia (NUD) are in many cases initiated by, or correlated to, inflammatory processes, where reactive oxygen species (ROS) play a crucial role. In this respect one prominent source of ROS are myeloperoxidase (MPO)-driven oxidation and chlorination reactions, assumed to be mainly responsible for tissue damage. In this study the contribution of the nine extracts to the overall performance of IG was compared with emphasis on MPO produced ROS. Concerning the influence on MPO dependent chlorination reactions, it turned out that of the nine IG-components Iberis amara extract (IAE) exerted the highest activity. Furthermore, this can impressively be reproduced in an ex vivo experiment with whole blood, where neutrophilic leukocytes are activated by zymosan. Moreover, along with the extract of chamomile flowers, IAE counteracts the pro-oxidative properties of caraway, peppermint and celandine. As a consequence. IG was also efficiently inhibiting MPO-catalysed chlorinations. As shown by the addition of catalase, the pro-oxidative effects of caraway, peppermint and celandine are due to their content of hydrogen peroxide. The latter is probably an autoxidation product of certain monoterpenes in the essential oil part of these extracts. If one of the component extracts of IG is omitted, the antioxidant acitivity is reduced. Thus we conclude that all the single extracts combined in IG are of importance for the therapeutical effect, working in concert. PMID- 15344844 TI - Synthesis and evaluation of the analgesic and anti-inflammatory activity of new 3(2H)-pyridazinone derivatives. AB - A series of 6-substituted-3(2H)-pyridazinone derivatives were synthesized and evaluated for analgesic and anti-inflammatory activities. The structures of these new pyridazinone derivatives were confirmed by their IR, 1H-NMR spectra and elementary analysis. Analgesic and anti-inflammatory activities of the title compounds have been evaluated. Four of the ten tested compounds possessed significant analgesic effects in the phenylbenzoquinone-induced writhing test (PBQ test). The most active derivatives 8a, 8b, 8d, 8e were void of gastric ulcerogenic effect or acute toxicity at the maximal dose (200 mg/kg p.o.). In the carrageenan-induced paw edema model, compound 8d (6- [4- (2-fluorophenyl) piperazin-1-yl]-3(2H)-pyridazinone) showed anti-inflammatory activity similar to that of the standard drug indometacin (CAS 53-86-1). A significant dependence of the anti-inflammatory effect on the substituents was observed; The pharmacological study of these compounds confirms that modification of the chemical group at position 6 of the 3(2H)-pyridazinone ring influences analgesic and anti-inflammatory activities. PMID- 15344845 TI - In vitro antispasmodic and anti-inflammatory effects of Drosera rotundifolia. AB - In investigations of the anti-inflammatory and spasmolytic effects of Drosera rotundifolia two extracts were tested in different in vitro assays. An aqueous and an ethanolic extract inhibited human neutrophil elastase, achieving IC50 values of 5 and 1 microg/mL, respectively. The very low naphthoquinone concentrations in the extracts seem not to be responsible for the effect, as the pure compounds were not effective in the test system used. Thus, flavonoids like hyperoside, quercetin and isoquercitrin, which were detected in the extracts in considerable concentrations, may contribute to the activity. These substances showed activity in the assay. Ellagic acid, detected especially in the ethanolic extract in higher amounts, was substantially less active than the flavonoids. In guinea-pig ileum the extracts led to an antispasmodic effect possibly by affecting an allosteric binding site of the muscarinic M3 receptors. PMID- 15344846 TI - Evaluation of anti-prion activity of congo red and its derivatives in experimentally infected hamsters. AB - Among transmissible spongiform encephalopathies (TSE), particularly dreadful are the bovine spongiform encephalopathy (BSE), because of its epidemic character, and the new variant of Creutzfeldt-lakob disease (vCJD) in man, possibly related to BSE prion, through the intake of infected food. To treat TSE, many potentially therapeutic agents have been tested: some of them, among which is Congo Red (CAS 573-58-0, CR), delayed the onset of symptoms in scrapie-infected rodents, and some CR derivatives proved to be effective in vitro. The capacity of a synthesized CR derivative (CR-A) and of the aromatic central benzidine rings of CR (CR-B) to abrogate scrapie-induced disease in experimentally infected hamsters was assayed. CR, used as reference substance, administered i.c. after pre incubation with the scrapie inoculum, was strongly effective in slowing the progression of the infection, while both CR-A and CR-B, administered alone or together, were not effective. Both CR-A and CR, when administered by subcutaneous route in i.c. scrapie-infected animals. prolonged the survival time in comparison to controls; CR-B was not effective. Moreover, both CR and CR-A were very effective in prolonging the survival time of i.p. scrapie-infected hamsters. The hypothesis of possible different mechanisms of interaction between CR or CR-A and the scrapie agent related to the chemical structures of the molecules is discussed. PMID- 15344847 TI - Efficacy of ivermectin in a patient population concomitantly infected with intestinal helminths and ectoparasites. AB - Ivermectin (CAS 70288-86-7) is a potent antiparasitic drug. However, studies have not been published evaluating the efficacy of ivermectin in a patient population concomitantly infected with intestinal helminths and ectoparasites. Here the results of an open trial on the efficacy of ivermectin in a heavily poly parasitized population in northeast Brazil are presented. Two hundred and fifty one patients were enrolled. Two doses of ivermectin (200 microg/kg) were given at an interval of 10 days. The cure rates for intestinal helminthiases were: 100% for strongyloidiasis and enterobiasis, 99% for ascariasis, 84% for trichuriasis, 68% for hookworm disease, and 50% for hymenolepiasis. Cure rates for ectoparasitoses were: 100% for cutaneous larva migrans, 99% for pediculosis, 88% for scabies and 64% for tungiasis. The results show that two doses of ivermectin are highly efficacious for most intestinal helminths and ectoparasites simultaneously present in an impoverished population. The drug seems to be particularly useful when polyparasitism is expected to occur or when public health measures aim to reduce both intestinal helminthiases and parasitic skin diseases by mass chemotherapy. PMID- 15344848 TI - Sound-evoked vestibulo-ocular reflexes (VOR) in trained monkeys. AB - Acoustic stimulation of the vestibular system has been well documented in humans and has been accepted as a useful tool to diagnose vestibular disorders. The goal of this study was to establish an awake and behaving primate model that might be useful for investigating the neural mechanisms underlying acoustic activation of the vestibular system. We recorded sound-evoked eye movements in monkeys while they performed ocular motor tasks. In the first part of the study, an acoustic click (1 ms, 99 to approximately 125 db peak SPL) was delivered to one of the monkeys' ears while they fixated on visual targets of varying eccentricities and viewing distances. Acoustic clicks were found to evoke well-defined biphasic eye velocity responses. For the movement in the horizontal direction, the first eye velocity peaks were always away from the stimulated ear. For the movement in the vertical direction, however, the directions of the first eye velocity peaks varied from monkey to monkey. This variability was difficult to interpret in the absence of torsional measurement. Thus, our analysis in this report was focused on horizontal eye movements. We found that click-evoked eye movements were disjunctive, with larger first horizontal eye velocity peaks from the eye ipsilateral to the stimulated ear (the amplitude ratio was 1.8 +/- 0.3, n=4). The amplitudes of the first horizontal peaks were also linearly correlated with gaze eccentricity and viewing distance. In the second part of the study, we found that a brief tone-pulse (100 ms, 125 db peak SPL) evoked eye movements that exhibited a well-defined frequency tuning with the most effective stimulating frequencies ranging from 1 K to 1.5 KHz. These data demonstrate that the sound-evoked eye movements in behaving monkeys are well defined and reproducible. This paradigm may be useful for studying the neural mechanisms underlying acoustic activation of the vestibular system. PMID- 15344849 TI - Role of lateral non-primary motor cortex in humans as revealed by epicortical recording of Bereitschaftspotentials. AB - In order to clarify the role of the lateral non-primary motor area in the control of voluntary movements, we studied movement-related cortical potentials (MRCPs) by direct epicortical recording from the lateral frontal lobe in nine patients with intractable partial epilepsy as a part of presurgical evaluation. We adopted movement tasks involving different body sites: eye closing, lip pursing, shoulder abduction, middle finger extension, thumb abduction, and foot dorsiflexion. We found that one or two small areas on the caudal lateral convexity of the frontal lobe generated pre-movement potential shifts regardless of the sites of movement (omni-Bereitschaftspotential; "omni-BP"). Such regions were located at or just rostral to the primary motor face area in six subjects, and at or rostral to the primary motor upper extremity area in three. Moreover, half of those areas were identified just adjacent (either rostral or caudal) to the primary negative motor area (PNMA), a cortical area of the lateral frontal lobe where negative motor responses were elicited by electric cortical stimulation. In conclusion, it is suggested that the lateral non-primary motor area plays a significant role, and has a close and direct relationship with other cortical areas in the frontal lobe, just like its counterpart on the mesial frontal cortex (supplementary negative motor area, SNMA). PMID- 15344850 TI - Adaptive changes in vergence eye movements induced by vergence-vestibular interaction training in monkeys. AB - Clear vision of objects moving in three-dimensional space near an observer is attained by a combination of smooth-pursuit and vergence eye movements. The two systems must interact with the vestibular system to maintain the image of the object on the fovea. Previous studies showed that training with smooth-pursuit vestibular interactions resulted in adaptive changes in the smooth-pursuit response. Although vergence and smooth-pursuit systems are thought to have separate neural substrates, recent studies indicate that the caudal parts of the frontal eye fields that receive vestibular inputs contain neurons that discharge in response to combinations of smooth-pursuit and vergence. This combination of discharge sensitivities suggests the possibility that adaptive changes may be induced in the vergence system by vestibular inputs during vergence-pursuit training. To explore this possibility, we examined the effects of training with conflicting vestibular and vergence tracking in four head-stabilized monkeys. Animals were rewarded for tracking a laser spot that moved towards or away from them at 1 Hz in phase with sinusoidal whole-body rotation (+/- 5 degrees) in the pitch plane; the spot moved closer when the monkey's nose moved downward. From the monkey's point of view, the spot moved sinusoidally 10-66 cm in front of them along the mid-sagittal plane, requiring symmetrical vergence eye movements of 4.8 degrees for each eye. Eye movements induced by equivalent spot motion at 0.3-1.0 Hz with or without chair rotation were examined before and after training for each session (0.5-1.0 h). Before training, pitch rotation alone in complete darkness did not induce vergence eye movements in any of the monkeys tested. Vergence tracking without chair rotation showed decreased gain and increased phase lag (re vergence target velocity) at frequencies above 0.5 Hz. After training, the vergence response during chair rotation with the spot showed significantly higher gains and smaller phase lags at 0.3-1.0 Hz in all monkeys. Pitch rotation alone in complete darkness induced vergence eye movements with gains (eye vergence/chair) of 0.15-0.35 after training in two monkeys. These results suggest that vestibular information can be used effectively to modify vergence tracking. PMID- 15344851 TI - Saccade-vergence dynamics and interaction in children and in adults. AB - Peak velocity, duration and accuracy of eye movements (saccade, vergence and combined saccade-vergence eye movements) were investigated in fourteen normal children (4.5 to 12 years of age) and ten normal adults (22 to 44 years of age). Horizontal movements from both eyes were recorded simultaneously by the oculometer, a photoelectric device. Peak velocity of all eye movements, saccades, and vergence (convergence and divergence), attains adult levels by the age of 4.5 years and there is no significant change over the age range studied (4.5 to 44 years). Vergence duration is longer only in young children (below 8 years of age). The reciprocal interaction between saccade and vergence during combined movements known in adults, i.e. acceleration of the vergence by the saccade (increase of velocity and decrease of duration) and deceleration of the saccade by the vergence (decrease of velocity and increase of duration) was found to be similar in children. The accuracy of eye movements is good on average for both saccades and vergence by the age of 4.5 years, and does not change with age; an exception is the variability of saccade amplitude, which is higher in children less than 8 years old. Taken together, the results indicate early maturation of brainstem structures controlling spatio-temporal aspects of saccades, vergence and their interaction. PMID- 15344852 TI - Endomorphins exit the brain by a saturable efflux system at the basolateral surface of cerebral endothelial cells. AB - Endomorphin-1 (EM-1) and endomorphin-2 (EM-2) are two highly selective mu-opiate receptor agonists. We recently demonstrated that EM-1 and EM-2 have a saturable transport system from brain-to-blood in vivo. Since the endothelial cells are the main component of the non-fenestrated microvessels of the blood-brain barrier (BBB), we examined whether these endogenous tetrapeptides have a saturable transport system in cultured cerebral endothelial cells. EM-1 and EM-2 binding and transport were studied in a transwell system in which primary mouse endothelial cells were co-cultured with rat glioma cells. We found that binding of both endomorphins was greater on the basolateral than the apical cell surface. Flux of EM-1 and EM-2 occurred predominantly in the basolateral to apical direction, each showing self-inhibition with an excess of the respective endomorphin. Transport was not influenced by the addition of the P-glycoprotein inhibitor, cyclosporin A. Neither the mu-opiate receptor agonist DAMGO nor the delta-opiate receptor agonist DPDPE had any effect on the transport. Thus, the results show that a saturable transport system for EM-1 and EM-2 occurs at the level of endothelial cells of the BBB, and unlike beta-endorphin and morphine, P glycoprotein is not needed for the brain-to-blood transport. Cross-inhibition of the transport of each endomorphin by the other suggests a shared transport system that is different from mu- or delta-opiate receptors. As endormorphins are mainly produced in the CNS, the presence of the efflux system at the BBB could play an important role in pain modulation and neuroendocrine control. PMID- 15344853 TI - Anticipatory attention to verbal and non-verbal stimuli is reflected in a modality-specific SPN. AB - The time estimation paradigm allows the recording of anticipatory attention for an upcoming stimulus unconfounded by any anticipatory motor activity. Three seconds after a warning signal (WS) subjects have to press a button. A button press within a time window from 2,850 ms to 3,150 ms after the WS is considered 'correct', a movement prior to 2,850 ms after the WS is labelled 'too early' and a movement after 3,150 ms is labelled 'too late'. Two seconds after the button press a Knowledge of Results (KR) stimulus is presented, informing the subject about the correctness of the response. Stimulus Preceding Negativity (SPN) is a slow wave which is recorded prior to the presentation of the KR stimulus. The SPN has a right hemisphere preponderance and is based upon activity in a network in which prefrontal cortex, the insula Reili and the parietal cortex are crucial. In the present study we asked two questions: (1) does the SPN show modality specificity and (2) does the use of verbal KR stimuli influence the right hemisphere preponderance? Auditory and visual stimuli were presented, in a verbal mode and in a non-verbal mode. SPN amplitudes prior to visual stimuli were larger over the visual cortex than prior to auditory stimuli. SPN amplitudes prior to auditory stimuli were larger over the frontal areas than prior to visual stimuli. The use of verbal stimuli did not influence the right hemisphere preponderance. We concluded that apart from the supramodal effect of KR stimuli in general, there is (first) a modality-specific activation of the relevant sensory cortical areas. The supramodal network underlying the attention for and the use of KR information is activated either from different sensory areas or from language processing cortical areas. PMID- 15344854 TI - [Photopsies can have other causes than retinal detachment]. PMID- 15344855 TI - [Hypercoagulation and cancer. New treatment possibilities]. PMID- 15344856 TI - [Mitochondrial diabetes with a focus on maternally inherited diabetes and deafness]. PMID- 15344857 TI - [Should children with breath holding spells be treated with iron?]. PMID- 15344858 TI - [Motivational counseling. Possible in an emergency department?]. PMID- 15344859 TI - [Assessment of clinical factors associated with falls in a population of patients admitted to a geriatric department]. PMID- 15344860 TI - [Administration of medications. Joint charts for prescription--transcription and dispensing errors]. PMID- 15344861 TI - [Computer use and carpal tunnel syndrome: a 1-year follow-up study]. PMID- 15344862 TI - [Cohort study on effects of parathyroid surgery in primary hyperparathyroidism]. PMID- 15344863 TI - [Acalculous cholecystitis and acute pancreatitis after infection with Salmonella enteritidis]. PMID- 15344864 TI - [Melanoma-associated retinopathy in a patient without a primary tumour]. PMID- 15344865 TI - [Extreme hypercalcemic crisis in a patient with malignant lymphoma]. PMID- 15344866 TI - [Circulating endothelial stem cells: a new cardiovascular marker?]. PMID- 15344867 TI - [They promise one thing and promise to keep something else 2]. PMID- 15344868 TI - [Medical error!2]. PMID- 15344869 TI - [Colorectal surgeons--more debate!]. PMID- 15344870 TI - [Overinterpretation of a non-conclusive population study on breast cancer]. PMID- 15344871 TI - [Exposure of infants to outdoor and indoor tobacco smoke pollution]. PMID- 15344872 TI - [I'll have a referral]. PMID- 15344873 TI - [Is the Ethics Committee devoid of honesty?]. PMID- 15344874 TI - [War om the harbours and on the roads]. PMID- 15344875 TI - [Sexual relations between physicians and patients]. PMID- 15344876 TI - [Artificial insemination--what with the children?]. PMID- 15344877 TI - [One wonders...]. PMID- 15344878 TI - Thermodynamics of protein-ligand interactions: history, presence, and future aspects. AB - The understanding of molecular recognition processes of small ligands and biological macromolecules requires a complete characterization of the binding energetics and correlation of thermodynamic data with interacting structures involved. A quantitative description of the forces that govern molecular associations requires determination of changes of all thermodynamic parameters, including free energy of binding (deltaG), enthalpy (deltaH), and entropy (deltaS) of binding and the heat capacity change (deltaCp). A close insight into the binding process is of significant and practical interest, since it provides the fundamental know-how for development of structure-based molecular design strategies. The only direct method to measure the heat change during complex formation at constant temperature is provided by isothermal titration calorimetry (ITC). With this method one binding partner is titrated into a solution containing the interaction partner, thereby generating or absorbing heat. This heat is the direct observable that can be quantified by the calorimeter. The use of ITC has been limited due to the lack of sensitivity, but recent developments in instrument design permit to measure heat effects generated by nanomol (typically 10-100) amounts of reactants. ITC has emerged as the primary tool for characterizing interactions in terms of thermodynamic parameters. Because heat changes occur in almost all chemical and biochemical processes, ITC can be used for numerous applications, e.g., binding studies of antibody-antigen, protein peptide, protein-protein, enzyme-inhibitor or enzyme-substrate, carbohydrate protein, DNA-protein (and many more) interactions as well as enzyme kinetics. Under appropriate conditions data analysis from a single experiment yields deltaH, K(B), the stoichiometry (n), deltaG and deltaS of binding. Moreover, ITC experiments performed at different temperatures yield the heat capacity change (deltaCp). The informational content of thermodynamic data is large, and it has been shown that it plays an important role in the elucidation of binding mechanisms and, through the link to structural data, also in rational drug design. In this review we will present a comprehensive overview to ITC by giving some historical background to calorimetry, outline some critical experimental and data analysis aspects, discuss the latest developments, and give three recent examples of studies published with respect to macromolecule-ligand interactions that have utilized ITC technology. PMID- 15344879 TI - Age-related changes in the properties of the endothelin receptor system at protein and mRNA levels in the rat vas deferens. AB - As age-related changes occur in the properties of the endothelin (ET) receptor system in several mammalian tissues, and as there are significant amounts of functional ET receptors in the vas deferens, we investigated the age-related changes in the ET receptor system at the protein and mRNA levels in the rat vas deferens. The ET system was investigated in the vasa deferentia of 3 weeks, 3 months and 22 months old rats. ET receptors were characterized and quantified at the protein level by radioligand receptor binding, and gene transcript levels of ET-1, ET-3, ET converting enzyme-1 (ECE-1), and ET(A) and ET(B) receptor subtypes were quantified by real-time reverse transcription polymerase chain reaction (RT PCR). The results of radioligand receptor binding assays demonstrate that there is a higher density of total ET receptors in the vas deferens of 3 weeks old rats than in 3 months and 22 months old rats, and that the predominant ET receptor is of the ET(A) subtype in all three ages. Real-time RT-PCR data show that the predominant mRNA expression of ETs and their receptors in all age groups studied are ET-1 and the ET(A) receptor subtype, respectively. Furthermore, ET-1, ET-3, ECE-1, and ET(A) and ET(B) receptor subtype mRNAs are expressed at higher levels in the 3 weeks old rats as compared with the other two age groups. These results demonstrate the presence of age-related changes in the properties of the ET receptor system at both protein and mRNA levels in the rat vas deferens. PMID- 15344880 TI - Inhibition of phosphatidylinositol-3-kinase enhances insulin stimulation of insulin receptor substrate 1 tyrosine phosphorylation and extracellular signal regulated kinases in mouse R- fibroblasts. AB - Insulin stimulates phosphatidylinositol-3-kinase (PI3K) and extracellular signal regulated kinases (ERK) in various mammalian cells. To study the role of PI3K in insulin stimulation of ERK, we employed PI3K inhibitor LY294002 and mouse embryonic R- fibroblasts lacking IGF-1 receptors. In these R- cells, PI3K inhibition by LY294002 enhanced insulin stimulation of ERK phosphorylation whereas LY294002 inhibited insulin stimulation of Akt phosphorylation. The enhanced insulin stimulation of ERK phosphorylation was accompanied by increased IRS-1 tyrosine phosphorylation. Insulin stimulation of insulin receptor tyrosine phosphorylation was not altered. PI3K inhibition increased IRS-1-Grb2 complex formation and ras activity following insulin treatment of cells. Increased insulin stimulation of ERK by PI3K inhibition was mediated by the MEK/ERK pathway, but did not involve inhibitory Ser259 phosphorylation of raf that was reported to be mediated by Akt. In summary, PI3K inhibition in R- cells enhanced insulin stimulation of ERK phosphorylation by mechanisms involving enhancement of IRS-1 tyrosine phosphorylation, IRS-1-Grb2 complex formation and the ras/MEK/ERK pathway. PMID- 15344881 TI - Signaling pathways of the F11 receptor (F11R; a.k.a. JAM-1, JAM-A) in human platelets: F11R dimerization, phosphorylation and complex formation with the integrin GPIIIa. AB - The F11 receptor (F11R) (a.k.a. Junctional Adhesion Molecule, JAM) was first identified in human platelets as a 32/35 kDa protein duplex that serves as receptor for a functional monoclonal antibody that activates platelets. We have sequenced and cloned the F11R and determined that it is a member of the immunoglobulin (Ig) superfamily of cell adhesion molecules. The signaling pathways involved in F11R-induced platelet activation were examined in this investigation. The binding of M.Ab.F11 to the platelet F11R resulted in granule secretion and aggregation. These processes were found to be dependent on the crosslinking of F11R with the Fc gammaRII by M.Ab.F11. This crosslinking induced actin filament assembly with the conversion of discoidal platelets to activated shapes, leading to the formation of platelet aggregates. We demonstrate that platelet secretion and aggregation through the F11R involves actin filament assembly that is dependent on phosphoinositide-3 kinase activation, and inhibitable by wortmannin. Furthermore, such activation results in an increase in the level of free intracellular calcium, phosphorylation of the 32 and 35 kDa forms of the F11R, F11R dimerization coincident with a decrease in monomeric F11R, and association of the F11R with the integrin GPIIIa and with CD9. On the other hand, F11R-mediated events resulting from the binding of platelets to an immobilized surface of M.Ab.F11 lead to platelet adhesion and spreading through the development of filopodia and lammelipodia. These adhesive processes are induced directly by interaction of M.Ab.F11 with the platelet F11R and are not dependent on the Fc gammaRII. We also report here that the stimulation of the F11R in the presence of nonaggregating (subthreshold) concentrations of the physiological agonists thrombin and collagen, results in supersensitivity of platelets to natural agonists by a F11R-mediated process independent of the Fc gammaRII. The delineation of the two separate F11R-mediated pathways is anticipated to reveal significant information on the role of this cell adhesion molecule in platelet adhesion, aggregation and secretion, and F11R-dependent potentiation of agonist-induced platelet aggregation. The participation of F11R in the formation and growth of platelet aggregates and plaques in cardiovascular disorders, resulting in enhanced platelet adhesiveness and hyperaggregability, may serve in the generation of novel therapies in the treatment of inflammatory thrombosis, heart attack and stroke, and other cardiovascular disorders. PMID- 15344882 TI - [Apoptosis and heart disorders]. AB - Papers on the significance of cardiomyocyte apoptosis in cardiovascular disorders development are reviewed. Some distinctive features of regulation and morphological manifestations of cardiomyocyte apoptosis are described. Although a great majority of cardiomyocytes are highly differentiated cells, apoptosis related loss of cellular weight is noted in the number of pathological processes. Data on the role of apoptosis in myocardial ischemia in cardiomyopathies and in heart failure are presented. PMID- 15344883 TI - [Antioxidants-induced rearrangements of actin cytoskeleton in 3T3 and 3T3-SV40 fibroblasts]. AB - Effect of antioxidants on actin cytoskeleton in 3T3 fibroblasts and 3T3 fibroblasts transformed with SV40 virus (3T3-SV40 cells) was studied. Antioxidants used were as follows: N-acetyl-L-cysteine (NAC), (-)-2-oxo-4 thiazolidine-carboxylic acid (OTZ), and glutathione in the reduced form (GSH). Both NAC and OTZ are precursors of GSH in the cell, but, in contrast to NAC, OTZ reduces inside the cell forming L-cysteine. The presence of NAC (5-20 mM) in the culture medium of both cell types resulted in loosening of monolayer, fragmentation of stress fibers, and the appearance of amorphous actin structures. As 3T3-SV40 cells contain less actin stress fibers than 3T3 cells, the NAC induced rearrangements of actin cytoskeleton were stronger in these cells than in 3T3 cells. In contrast to NAC, OTZ (10-20 mM) did not destroy monolayer and did not induce any visible disappearance of stress fibers either in 3T3 or 3T3-SV40 cells. However, in the presence of OTZ, amorphous actin-containing structures were observed in 3T3-SV40 cells. The effect of glutathione on both cell types was similar to that of NAC. The time required for GSH-induced alterations of actin cytoskeleton (about 5 h) was consistent with the increase in the intracellular level of reactive oxygen species (4 h after addition of GSH to the culture medium). Upon removal of the antioxidants from the medium, actin filament structures were reconstructed. However, within 24 h after withdrawal of NAC or GSH, only a partial reconstruction of stress fibers was observed in 3T3 cells. On the contrary, 3T3-SV40 cells demonstrated formation of well-structured actin fibers similar to normal fibroblasts. These results suggest that GSH can act as a pro-oxidant in the absence of oxidative stress. PMID- 15344884 TI - ["Dark" and "pale" cells of the amygdala complex in dynamics of the estrous cycle]. AB - Data on cytological peculiarities of dorsomedial nucleus neurones of the amygdala complex, one of the main zones of sexual dimorphism, in dynamics of estrous cycle are reported. We show that structural and functional characteristics of "dark" and "pale" cells may change depending on the concentrations of gonadal steroids in estrous and metaestrous stages. This specifies the previous hypothesis about mutual reorganization of these cells. PMID- 15344885 TI - [Regeneration of white rat liver during cholestasis]. AB - Cholestasis inhibits expression of early response genes in rat hepatocytes stimulated to proliferation and restricts the process related to reparative regeneration. The occlusion of general bile duct induces bile acid accumulation, which suppresses 11-beta-hydroxysteroid-dehydrogenase activity. Hence, the concentration of corticosterone is increased, which suppresses the expression of early response genes in rat hepatocytes. The restoration of liver mass occurs due to proliferation of reserved cells. An unusual mitotic peak of these cells is registered 34 h after the operation. PMID- 15344886 TI - [Dynamics of structural and functional changes in wheat root cells under the action of protonophore]. AB - Structural and functional changes in wheat root cells during long-term action of a protonophore--carbonyl cyanide 3-chlorophenylhydrazone (CCCP)--were studied. It was demonstrated that CCCP affected the electrical potential and inward resistance of cells, increased K+ ions release to the incubation medium, inhibits oxygen uptake for 1-4 h, which was followed by oxygen uptake stimulation for 6 h of treatment. These changes of physiological processes were accompanied with a variety of ultrastructural changes in cell organization, namely in the structure of mitochondria, endoplasmic reticulum canals, and the nucleus. The role of protons is discussed, in particular, in the regulation of metabolic state of mitochondria, and in general regulation of structural and functional conditions of cells. PMID- 15344887 TI - [Degradation of Cdc25A phosphatase in HeLa cells under normal and stress conditions]. AB - Cdc25A phosphatase regulates cell cycle progression by removing the inhibitory phosphates from cyclin-dependent kinases. Activity of Cdc25A depends on its phosphorylation status. During normal cell cycle progression and after DNA damage phosphorylation by Chk1 (or Chk2) triggers Cdc25A degradation via ubiquitin proteasome pathway. In this study we investigate the role of various phosphorylation sites (Ser123, Ser75, Ser17 and Ser115) in the regulation of Cdc25A stability. We have shown that only S75A mutation abrogates Cdc25A degradation both in normal and stress conditions. We also studied the influence of stable form of Cdc25A on checkpoint progression after DNA damage. We have found out that delay in DNA synthesis after UV and IR does not depend on Cdc25A activity. However, the presence of stable Cdc25A increases the number of mitotic cells after these stresses. PMID- 15344888 TI - [The role of stress-kinase p38 in cell cycle blocking delay under osmotic stress]. AB - Alteration in osmotic conditions leads to activation of some defensive mechanisms resulting in blocking the cell cycle progression. One of stress kinases, activated after osmotic stress, is p38 MAPK. In the present study the role of p38 both in intra-S-phase and G2/M checkpoint is shown. Besides, p38-dependent degradation of Cdc25A phosphatase after osmotic stress is demonstrated. Expression of stable form of Cdc25A results in a partial abrogation of G2/M block of the cell cycle, but has no effect on DNA synthesis arrest. PMID- 15344889 TI - Small stable RNA level depends on the physiological state of the cell. AB - The level of 5S rRNA and tRNAi(Met)1 synthesized by RNA polymerase III was investigated in human epidermoid carcinoma cells A431 at different physiological states: low and high proliferation and apoptosis. The real-time RT-PCR method using SYBR Green I was applied to measure certain RNA species in total cellular RNA. The share of 5S rRNA was practically the same in slowly and actively proliferating A431 cells, but increased about 2.5-fold in apoptotic cells. The share of initiator tRNAi(Met)1 in actively proliferating and apoptotic cells was 1.5-2.0 times higher than in slowly proliferating cells. Our results suggest a possible existence of special mechanisms regulating RNA polymerase III-directed transcription from different type promoters in accordance with the physiological state of the cell. PMID- 15344890 TI - [Purification and ligand-receptor analysis of insulin-related peptides from pedal ganglion of the mollusc Anodonta cygnea]. AB - Six insulin-related peptides (IRPs) from pedal ganglions of the molluscs Anodonta cygnea have been isolated and purified by reverse-phase chromatography. Each peptide (designated as IRP8-IRP13) showed its own retention time on the HPLC column. The testing of IRPs in radioreceptor systems specific for insulin and insulin growth factor-I (IGF-I) showed their ability to bind to both types of receptors. The concentration of IRPs, producing a 50% inhibition of porcine 125I insulin binding with rat liver plasma membrane receptors (IC50) for IRP 10, was 1167 nM, IRP11--833 nM, IRP13--1333 nm. IRP8, IRP9, IRP12 in the maximum concentration of 10(4) ng/ml displaced less than 50% of labeled hormone. All of the six peptides were capable of competing with human 125I-IGF-I for binding to receptors of a fraction of rat brain membranes. IRP8, IRP9 and IRP12 had close means equal to 1167 nM, 1500 nM, 1167 nM, respectively. Another group including IRP10, IRP11 and IRP13 showed a much higher activity (833, 83 and 500 nM, respectively). The results obtained from radioligand analysis revealed the predominance of IGF-I binding properties in all peptides of pedal ganglions. At the same time, apparent proximity of IRP's physico-chemical characteristics to porcine insulin, and also the revealed dose-dependent binding to both insulin and IGF-I receptors suggest a bifunctionality of mollusc peptides. The expression level of this bifunctionality may be associated with the molecular structure pecularities of individual isoforms. PMID- 15344891 TI - [Immunocytochemical localization of atrial natriuretic peptide in endothelial and granular cells of the heart of lophotrochozoa]. AB - In parallel with contraction, vertebrate cardiomyocytes perform endocrine function and produce natriuretic peptides (NP)--ANP and BNP--involved in cardiovascular homeostasis maintenance. ANP-like peptides have been reported also in hearts of some invertebrates, however, their cellular localization was not determined. By electron microscopical immunocytochemistry with polyclonal monospecific antibodies raised against ANP and protein A-gold technique, we have localized ANP-like immunoreactivity in granules within endothelial cells in the heart of the brachiopod Rhynchonella psittacea, the polychaete Arenicola marina, and the gastropod mollusc Achatina fulica--all being representatives of the major phylogenetic group Lophotrochozoa. ANP-like immunoreactivity was also revealed in one of 3 morphologically distinguishable types of granules in the snail heart granular cells. By electron microscopical autoradiography with the use of [3H] thymidine, the ability for DNA synthesis was demonstrated in heart endothelial cells of the investigated animals. Forms of NP-system organization in hearts of Lophotrochozoa and Vertebrates, and close histogenetic relationships of endothelial and granular cells in the snail heart are discussed. PMID- 15344892 TI - [Dynamics of structural changes of Paramecium caudatum and Bursaria truncatella macronuclear chromatin and nucleoli under hypotonic conditions]. AB - Dynamics of structural changes of nucleoli, complex nucleolar aggregates and chromatin bodies in macronuclei (Ma) of ciliates Paramecium candatum and Bursaria truncatella under hypotonic conditions was studied. It was shown that after a 3 min hypotonic treatment nuclei swelled and became highly vacuolated. 3D reconstruction showed that such nucleoli were formed by nucleolonema-like threads about 100-200 nm in thickness. Intranucleolar chromatin bodies decompacted, but remained bound with fibrillar component of the nucleolus by thin fibres about 10 nm thick. After 6 min hypotonic treatment the nucleolar material loosened and had a "gauze", or network-like appearence. After 10 min hypotonic treatment nucleoli dissociated completely. It was shown that a transition of chromatin bodies from completely compact to partially and fully decompacted state occurred cooperatively in different regions of Ma. In particular, chromatin bodies in the central part of complex nucleolar aggregates decompacted much faster than those in the Ma karyoplasm. It evidences for a specific, well-ordered chromatin organization in Ma. Prolonged hypotonic treatment led to a complete dissociation of Ma components; fibres 6-10 nm thick were solely observed in such preparations. Such fibres may represent remnant structures of the nuclear matrix. Dynamics of Ma chromatin bodies decompaction correlates well with that of chromomeres in the nuclei of higher eukaryotes. Our data confirm that chromatin 100-200 nm bodies in the ciliate Ma are analogues of chromomeres--looped discrete chromatin domains, observed in the nuclei of higher eukaryotes. PMID- 15344893 TI - [The role of iron-regulated genes in microbial pathogenesis]. AB - An important component of nonspecific defence of human or animal organisms against microbial invasion is attempt to withhold growth-essential iron from invading bacteria. To overcome the barrier bacterial iron uptake systems is derepressed at low iron concentrations. In many pathogenic and nonpathogenic bacteria the regulation of the corresponding genes depends on the regulatory protein Fur. For cloning Fur-regulated loci, methods based on affinity of Fur to the corresponding promoter sites are used. PMID- 15344894 TI - Spontaneous intracerebral hemorrhage: a review. AB - Spontaneous intracerebral hemorrhage (SICH) is a blood clot that arises in the brain parenchyma in the absence of trauma or surgery. This entity accounts for 10 to 15% of all strokes and is associated with a higher mortality rate than either ischemic stroke or subarachnoid hemorrhage. Common causes include hypertension, amyloid angiopathy, coagulopathy, vascular anomalies, tumors, and various drugs. Hypertension, however, remains the single greatest modifiable risk factor for SICH. Computerized tomography scanning is the initial diagnostic modality of choice in SICH, and angiography should be considered in all cases except those involving older patients with preexisting hypertension in thalamic, putaminal, or cerebellar hemorrhage. Medical management includes venous thrombosis prophylaxis, gastric cytoprotection, and aggressive rehabilitation. Anticonvulsant agents should be prescribed in supratentorial SICH, whereas the management of hypertension is controversial. To date, nine prospective randomized controlled studies have been conducted to compare surgical and medical management of SICH. Although definitive evidence favoring surgical intervention is lacking, there is good theoretical rationale for early surgical intervention. Surgery should be considered in patients with moderate to large lobar or basal ganglia hemorrhages and those suffering progressive neurological deterioration. Elderly patients in whom the Glasgow Coma Scale score is less than 5, those with brainstem hemorrhages, and those with small hemorrhages do not typically benefit from surgery. Patients with cerebellar hemorrhages larger than 3 cm, those with brainstem compression and hydrocephalus, or those exhibiting neurological deterioration should undergo surgical evacuation of the clot. It is hoped that the forthcoming results of the International Surgical Trial in IntraCerebral Hemorrhage will help formulate evidence-based recommendations regarding the role of surgery in SICH. PMID- 15344895 TI - Update on management of intracerebral hemorrhage. AB - Intracerebral hemorrhage (ICH) is a lingering cause of significant mortality and morbidity rates in contemporary society. Despite its established burden, considerably less investigative attention has been devoted to the study of ICH than other forms of stroke. Only a limited number of clinical studies have been performed to examine the surgical (both craniotomy and minimally invasive) and medical management of patients with ICH. No consistently efficacious strategies have been identified through such investigations. Limitations in study design and execution have universally impaired the interpretation and impact of available data. Management of ICH unfortunately remains heterogeneous across institutions, and it continues to suffer from the lack of proven medical and surgical effectiveness. Urgently needed are further prospective randomized controlled trials in which investigators consider the shortcomings of previous endeavors in the management of ICH. In the present article the authors review the current management practices of ICH, discuss the controlled trials, and highlight recent trials and future avenues of further study. PMID- 15344896 TI - Spontaneous intracerebral hemorrhage due to coagulation disorders. AB - Although intracranial hemorrhage accounts for approximately 10 to 15% of all cases of stroke, it is associated with a high mortality rate. Bleeding disorders account for a small but significant risk factor associated with intracranial hemorrhage. In conditions such as hemophilia and acute leukemia associated with thrombocytopenia, massive intracranial hemorrhage is often the cause of death. The authors present a comprehensive review of both the physiology of hemostasis and the pathophysiology underlying spontaneous ICH due to coagulation disorders. These disorders are divided into acquired conditions, including iatrogenic and neoplastic coagulopathies, and congenital problems, including hemophilia and rarer diseases. The authors also discuss clinical features, diagnosis, and management of intracranial hemorrhage resulting from these bleeding disorders. PMID- 15344897 TI - Intracerebral hematoma from aneurysm rupture. AB - OBJECT: Patients who present with an intraparenchymal hematoma associated with a ruptured aneurysm usually require urgent clot evacuation and aneurysm obliteration. The impact of the presence of hematoma on outcome has been poorly characterized. The authors report on 460 patients who had dense subarachnoid hemorrhage (SAH) (Fisher Grades 3 and 4) with and without associated hematoma. METHODS: Of the 959 consecutive patients who presented with SAH, 460 patients with Fisher Grade 3 and 4 SAH were analyzed and divided into two groups: those with (Group 1) and those without (Group 2) hematoma. The presenting Hunt and Hess grade and 6-month outcomes of the two groups were compared. Of the 460 patients, 116 (25%) had intraparenchymal hematomas and admission Hunt and Hess grades were worse in Group 1 compared with Group 2. Outcome scores were worse for Group 1 compared with Group 2; however, when comparing Group 1 and Group 2 within the same initial Hunt and Hess score, there was no statistical difference in outcome. CONCLUSIONS: Intraparenchymal hematoma in association with SAH does not differ significantly from those patients without associated hematomas. We therefore recommend aggressive clot evacuation and aneurysm obliteration. PMID- 15344898 TI - Neonatal venous cerebral hemorrhage. Report of two cases. AB - Intracranial pathological changes can occur as a result of impaired craniocervical venous return. Thrombosis of central venous access catheters was demonstrated in two neonates born at 38 and 27 weeks' gestation. Neither infant developed hemorrhage of prematurity as confirmed on cranial ultrasonography. Clinical evidence of vena cava thrombosis and associated spontaneous intraventricular hemorrhage developed on Day 24 and 36, respectively, and these findings were confirmed on imaging studies. In one infant the hemorrhage was accompanied by communicating hydrocephalus. The cause of the intracranial disease was attributable to the retrograde cerebral venous congestion. This, together with the primitive venous bed developing in the periventricular region, was associated with the spontaneous hemorrhage in the region of the foramen of Monro. To the authors' knowledge, this is the first report in the English-language literature of spontaneous neonatal intracerebral hemorrhage, due to thrombosis of the superior or inferior vena cava. The natural history of this condition is resolution without sequelae after appropriate therapeutic intervention for the vena cava thrombosis. PMID- 15344899 TI - Reliability assessment of computerized tomography scanning measurements in intracerebral hematoma. AB - OBJECT: As one of the aspects of the International Surgical Trial in Intracerebral Haemorrhage (STICH), prerandomization computerized tomography (CT) scans were collected. In the present study the authors determined the inter-and intraobserver variability of various parameters pertinent to CT scans obtained in patients with intracerebral hematomas (ICHs). METHODS: A protocol was devised to analyze CT scans in a uniform and systematic manner. Each observer evaluated the same set of scans twice, with a minimum 2-month interval between assessments. In addition to noting the side and the sites of involvement, the observers measured the scale present on the scan itself and the length, breadth, height, and depth of the spontaneous ICH as well as the midline shift. The intraclass correlation was very high (kappa 0.8-1) for the measurements of volume, depth, and midline shift. Good interobserver agreement (kappa 0.8-1) was demonstrated with regard to involvement of basal ganglia or thalamus, presence of intraventricular extension, and the side of the hematoma. Agreement was substantial (kappa 0.61-0.8) with regard to identifying primary involvement of particular lobes. Agreement was moderate (kappa 0.41-0.6) on the presence or absence of hydrocephalus. When comparing the first and the second sets of readings, the intraobserver agreement was good (80-100%). CONCLUSIONS: The study quantifies the degree of inter- and intraobserver agreement regarding evaluation of CT scans in patients with ICH when conducted in accordance with a set protocol. PMID- 15344900 TI - Endovascular strategies for carotid cavernous and intracerebral dural arteriovenous fistulas. AB - Interventional neuroradiology is a less invasive modality than open neurosurgery for the treatment of a wide range of neurovascular disorders. Refinements in technique and improvements in the design of microcatheters, guide wires, and embolic materials have yielded superior clinical outcomes and provided an impetus to treat more of these lesions endovascularly rather than surgically. Endovascular therapy is the standard of care for direct and indirect carotid artery cavernous sinus fistulas and may also be curative for dural arteriovenous fistulas (AVFs) in other locations. The authors provide review of the clinical features, pathophysiology, and management of intracranial AVFs focusing on contemporary endovascular treatment options. PMID- 15344901 TI - Malignant meningioma of the lateral ventricle. Case report. AB - This 65-year-old man presented with a very rare malignant meningioma in the trigonum of the right lateral ventricle. Neurological examination showed bilateral papilledema. Magnetic resonance imaging revealed a solid, enhancing tumor in the right trigonum with a hypointense cystic component located in the center of the tumor. The lesion was totally resected via a superior parietooccipital transcortical approach. Histological examination showed an anaplastic (malignant) meningioma with architectural disarray, high mitotic activity (20/10 hpf), necrosis, and cytological atypism. As in our case, heterogeneous signal, due to necrotic tissue and frequently demonstrated on both T1- and T2-weighted sequences, is suggestive of an aggressive type of meningioma. PMID- 15344902 TI - Transcranial color-coded Doppler ultrasonography for evaluation of children with hydrocephalus. AB - OBJECT: Hydrocephalus is a common disease process. Transcranial color-coded Doppler (TCCD) ultrasonography is an accepted noninvasive method with which to quantify intracranial blood flow in adults and children. The authors studied the applications of TCCD ultrasonography and the alterations of the flow velocity of the cerebral arteries in children with hydrocephalus. METHODS: One hundred thirty five children were divided into three groups: Group 1 comprised 40 infants with asymptomatic hydrocephalus who had well-functioning ventriculoperitoneal (VP) shunts; Group 2 comprised 10 children with symptomatic hydrocephalus who had malfunctioning shunts that were replaced; and Group 3 was a control group of 85 healthy infants. All patients underwent sequential measurements of cerebral blood flow (CBF) velocities (systolic and diastolic velocities) and resistivity index (RI). One group of patients underwent functional tests (compression of the anterior fontanelle and CO2 vasoreactivity) to determine hemodynamic changes in cerebral circulation. A significant statistical change in RI measurements, end diastolic CBF velocity, and percentage of change in RI was shown in patients with malfunctioning shunts, and in infants with a well-functioning VP shunt vasomotor reactivity was severely reduced. CONCLUSIONS: Transcranial color-coded Doppler ultrasonography can be used to perform follow-up assessments of normal and malfunctioning shunts in children with hydrocephalus; the functional tests are a noninvasive tool for evaluating the cerebral compliance and the cerebral autoregulation in infants with hydrocephalus. The autoregulatory capacity may partly or completely be lost in cases of long-term shunt-treated hydrocephalus, and loss of cerebral vasoreactivity may be responsible for long-term deficits commonly observed in children, which help explain some of symptoms related to slit ventricles. PMID- 15344903 TI - Deferoxamine-induced attenuation of brain edema and neurological deficits in a rat model of intracerebral hemorrhage. AB - OBJECT: In the authors' previous studies they found that brain iron accumulation and oxidative stress contribute to secondary brain damage after intracerebral hemorrhage (ICH). In the present study they investigated whether deferoxamine, an iron chelator, can reduce ICH-induced brain injury. METHODS: Male Sprague-Dawley rats received an infusion of 100 microl of autologous whole blood into the right basal ganglia and were killed 1, 3, or 7 days thereafter. Iron distribution was examined histochemically (enhanced Perl reaction). The effects of deferoxamine on ICH-induced brain injury were examined by measuring brain edema and neurological deficits. Apurinic/apyrimidinic endonuclease/redox effector factor-1 (APE/Ref-1), a repair mechanism for DNA oxidative damage, was quantitated by Western blot analysis. Iron accumulation was observed in the perihematoma zone beginning 1 day after ICH. Deferoxamine attenuated brain edema, neurological deficits, and ICH induced changes in APE/Ref-1. CONCLUSIONS: Deferoxamine and other iron chelators may be potential therapeutic agents for treating ICH. They may act by reducing the oxidative stress caused by the release of iron from the hematoma. PMID- 15344904 TI - Cellular uptake and photocytotoxicity of glycoconjugated porphyrins in HeLa cells. AB - Thirty-two glycoconjugated porphyrins were synthesized by a modification of Lindsey method in the presence of Zn(OAc)(2).2H(2)O as a template. The Zn(2+) ion template strategy improved the yield about three-fold in the case of meta substituted tetraphenylporphyrins. In addition, free-base porphyrins were obtained almost quantitatively by demetalation with 4 M HCl. Sixteen deacetylated glycoconjugated porphyrins were tested as candidate photodynamic therapy (PDT) drugs using HeLa cells. Most of the deacetylated glycoconjugated porphyrins showed higher cellular uptake than tetraphenylporphyrin tetrasulfonic acid (TPPS), and 5,10,15,20-tetrakis[4-(beta-D-arabinopyranosyloxy)phenyl]porphyrin (p 5d) in particular showed 18.5-fold higher uptake than TPPS. The photocytotoxicity of 5,10,15,20-tetrakis[4-(beta-D-glucopyranosyloxy)phenyl]porphyrin (p-5a), p-5d and TPPS was examined with HeLa cells, using a light dose of 16 J/cm(2). These photosensitizers had no cytotoxicity in the dark, but their photocytotoxicity increased in the order of TPPS < p-5a < p-5d. These results suggest p-5d is a good candidate for a PDT drug. PMID- 15344905 TI - Enhancement of endotoxin neutralization by coupling of a C12-alkyl chain to a lactoferricin-derived peptide. AB - Antibacterial peptide acylation, which mimics the structure of the natural lipopeptide polymyxin B, increases antimicrobial and endotoxin-neutralizing activities. The interaction of the lactoferricin-derived peptide LF11 and its N terminally acylated analogue, lauryl-LF11, with different chemotypes of bacterial lipopolysaccharide (LPS Re, Ra and smooth S form) was investigated by biophysical means and was related to the peptides' biological activities. Both peptides exhibit high antibacterial activity against the three strains of Salmonella enterica differing in the LPS chemotype. Lauryl-LF11 has one order of magnitude higher activity against Re-type, but activity against Ra- and S-type bacteria is comparable with that of LF11. The alkyl derivative peptide lauryl-LF11 shows a much stronger inhibition of the LPS-induced cytokine induction in human mononuclear cells than LF11. Although peptide-LPS interaction is essentially of electrostatic nature, the lauryl-modified peptide displays a strong hydrophobic component. Such a feature might then explain the fact that saturation of the peptide binding takes place at a much lower peptide/LPS ratio for LF11 than for lauryl-LF11, and that an overcompensation of the negative LPS backbone charges is observed for lauryl-LF11. The influence of LF11 on the gel-to-liquid-crystalline phase-transition of LPS is negligible for LPS Re, but clearly fluidizing for LPS Ra. In contrast, lauryl-LF11 causes a cholesterol-like effect in the two chemotypes, fluidizing in the gel and rigidifying of the hydrocarbon chains in the liquid-crystalline phase. Both peptides convert the mixed unilamellar/non lamellar aggregate structure of lipid A, the 'endotoxic principle' of LPS, into a multilamellar one. These data contribute to the understanding of the mechanisms of the peptide-mediated neutralization of endotoxin and effect of lipid modification of peptides. PMID- 15344906 TI - Regulation of stability of cyclin-dependent kinase CDK11p110 and a caspase processed form, CDK11p46, by Hsp90. AB - CDK11p110 (cyclin-dependent kinase 11p110, formerly known as PITSLRE) is a member of the CDK superfamily. It associates with cyclin L and is involved in the regulation of transcription and in premRNA splicing. During staurosporine-, Fas- and tumour necrosis factor a-induced apoptosis, CDK11p110, is cleaved by caspases to generate smaller 46-50 kDa proteins containing the catalytic kinase domain. Ectopic expression of the caspase-processed form CDK11p46 induces apoptosis. The mechanisms that regulate activation and stability of CDK11 isoforms are still unclear. In the present study, we demonstrate that in human melanoma cells CDK11p110 and CDK11p46 interact with Hsp90 (heat-shock protein 90) and its co chaperone cdc37. Furthermore, we show that the treatment of cells with the Hsp90 specific inhibitor geldanamycin leads to ubiquitination and enhanced degradation of both CDK11p110 and CDK11p46 through a proteasome-dependent pathway. We also determined that geldanamycin-triggered degradation of CDK11p46 slows down the progression of apoptosis. These results indicate that Hsp90 and cdc37 stabilize CDK11 kinase, and suggest that this stabilization is crucial for its pro apoptotic function. PMID- 15344907 TI - Identification of a novel rat hepatic gene induced early by insulin, independently of glucose. AB - We used mRNA differential display to identify new genes induced early after exposure to insulin. Our screening strategy was based on the comparison of gene expression during the time course of insulin induction in the liver of 12-day-old suckling rats both in vivo and in vitro. A novel, early induced transcript, EIIH, was identified that encodes a 353-amino-acid protein with several features suggesting that it may be secreted or bound to membranes. EIIH is also distantly related to a variety of LRR (leucine-rich repeat) proteins. Insulin treatment increased EIIH mRNA levels in the hepatocytes of suckling, fasted adult and STZ (streptozotocin)-treated diabetic rats, where insulin was required to maintain the basal level of EIIH expression. EIIH expression was induced during the suckling/weaning transition, and remained detectable thereafter. Tissue distribution analysis in adult rats revealed a pattern of expression mainly in the liver, intestine and islets of Langerhans, closely following that of the Glut2 (glucose transporter 2), suggesting that it may play a role in carbohydrate metabolism. EIIH may be a primary target of the transcriptional regulation by insulin, and may therefore constitute a new model to study the mechanisms by which insulin acts on gene transcription. PMID- 15344908 TI - Molecular cloning and characterization of ZFF29: a protein containing a unique Cys2His2 zinc-finger motif. AB - We have cloned a gene, ZFF29 (zinc-finger protein of human fetal liver erythroid cells 29), from human fetal liver erythroid cells. Two types of mature mRNA were identified and designated ZFF29a and ZFF29b. In human genome the ZFF29 gene is on chromosome 9q, and the two forms are splice variants. There is a unique transcription start site, which predicts major mRNAs composed of 2485 bases for ZFF29a and 1801 bases for ZFF29b. The anticipated mRNAs were demonstrated in K562 cells, but not in any adult human tissues examined by Northern blotting. In the mouse, reverse transcription-PCR revealed that the ZFF29 mRNA is present in adult bone marrow and ovary at a higher level than in any other tissues examined. These findings suggest that ZFF29 proteins are expressed in embryonic/fetal erythroid tissues. The deduced polypeptide chains of ZFF29a and ZFF29b are composed of 306 and 350 amino acids respectively. A unique zinc-finger motif composed of two contiguous Cys(2)His(2)-type fingers is common to both forms of ZFF29. They are nuclear proteins and ZFF29b, but not ZFF29a, is an activator of erythroid gene promoters. PMID- 15344909 TI - Identification, purification and characterization of a novel human blood protein with binding affinity for prostate secretory protein of 94 amino acids. AB - PSP94 (prostate secretory protein of 94 amino acids), an abundant protein within semen, has reported local functions within the reproductive tract and reported systemic functions. Mechanisms of action remain poorly understood, but binding to undefined molecules within the prostate, pituitary, testis and blood may initiate some of these actions. PSP94 serum measurements, especially of bound and free forms, have potential clinical utility in prostate cancer management. Identification of the binding molecules will help in the understanding of PSP94's action, and enable further development of PSP94 serum assays. PSPBP (PSP94 binding protein) was purified from human serum by ammonium sulphate fractionation, ion-exchange and affinity chromatography. The glycosylated protein ran as two bands on SDS/PAGE (70 and 95 kDa). N-terminal sequencing yielded a 30 amino-acid sequence, identical with the translated N-terminal region of a previously published cDNA (GenBank accession number AX136261). Reverse transcriptase PCR and plaque hybridization demonstrated PSPBP mRNA in peripheral blood leucocytes and in a prostate cDNA library. Northern blotting showed 2 kb mRNA species in prostate, testis, ovary and intestine. Immunohistochemistry demonstrated PSPBP in tissues, including pituitary and Leydig cells, supporting a role for PSP94 in hormonal control at the pituitary gonadal axis. ELISA demonstrated that PSPBP levels were significantly lower (P=0.0014) in the serum of a prostate cancer population (n=65) compared with a control population (n=70). PSPBP identification will help the understanding of PSP94's functions and facilitate ELISA development to address the clinical value of PSP94 serum assays. PMID- 15344910 TI - Cyclo-oxygenase type 2-dependent prostaglandin E2 secretion is involved in retrovirus-induced T-cell dysfunction in mice. AB - MAIDS (murine AIDS) is caused by infection with the murine leukaemia retrovirus RadLV-Rs and is characterized by a severe immunodeficiency and T-cell anergy combined with a lymphoproliferative disease affecting both B- and T-cells. Hyperactivation of the cAMP-protein kinase A pathway is involved in the T-cell dysfunction of MAIDS and HIV by inhibiting T-cell activation through the T-cell receptor. In the present study, we show that MAIDS involves a strong and selective up-regulation of cyclo-oxygenase type 2 in the CD11b+ subpopulation of T- and B-cells of the lymph nodes, leading to increased levels of PGE2 (prostaglandin E2). PGE2 activates the cAMP pathway through G-protein-coupled receptors. Treatment with cyclo-oxygenase type 2 inhibitors reduces the level of PGE2 and thereby reverses the T-cell anergy, restores the T-cell immune function and ameliorates the lymphoproliferative disease. PMID- 15344911 TI - Rhythmogenesis in vasopressin cells. AB - Many neurones in the central nervous system possess intrinsic pattern-generating properties, including vasopressin magnocellular neurosecretory cells. Synaptic input to vasopressin cells is not rhythmically patterned and yet these neurones fire action potentials in a 'phasic' activity pattern comprised of alternating periods of activity and silence that each last tens of seconds. This review describes the intrinsic and extrinsic mechanisms that generate phasic activity in vasopressin cells, highlighting recent work that has shown phasic activity to result from feedback modulation of synaptic inputs, and of intrinsic membrane properties, by peptides released from the dendrites of vasopressin cells. PMID- 15344912 TI - Activation of arylalkylamine N-acetyltransferase by phorbol esters in bovine pinealocytes suggests a novel regulatory pathway in melatonin synthesis. AB - In all mammalian species investigated, noradrenaline activates a beta adrenoceptor/cAMP/protein kinase A-dependent mechanism to switch on arylalkylamine N-acetyltransferase and melatonin biosynthesis in the pineal gland. Other compounds which are known to influence the melatonin-generating system are phorbol esters. The effect of phorbol esters on regulation of melatonin synthesis has been mainly investigated in rat pinealocytes. In these cells, phorbol esters do not increase cAMP levels and arylalkylamine N acetyltransferase on their own; however, phorbol esters potentiate the effects on cAMP and AANAT activity induced upon beta-adrenoceptor stimulation. In the present study, we investigated the effect of phorbol esters on the regulation of melatonin synthesis in bovine pinealocytes. We show that, in these cells, the phorbol esters 4beta-phorbol 12-myristate 13-acetate (PMA) or phorbol 12,13 dibutyrate have a direct stimulatory effect and induced 4-10-fold increases in AANAT protein levels, AANAT activity and melatonin production. The extent of these effects was similar to those induced by noradrenaline. Notably, responses to PMA were not accompanied by increases in cAMP levels. Northern blot analysis showed that Aanat mRNA levels did not change upon PMA treatment indicating that phorbol esters control AANAT at a post-transcriptional level. The effects on AANAT and melatonin production were reduced by use of protein kinase C inhibitors, but not by blockade of the cyclic AMP/protein kinase A pathway. Our results point towards a novel mechanism in the regulation of melatonin production that is cAMP-independent and involves protein kinase C. The study is of particular interest because regulation of melatonin biosynthesis in bovines may resemble that in primates more closely than that in rodents. PMID- 15344913 TI - Fos expression after mating in noradrenergic cells of the A1 and A2 areas of the medulla is altered by adrenalectomy. AB - In the female rat, the integrity of the ventral noradrenergic bundle (VNAB) is necessary to carry stimuli from the uterine cervix and vagina to brain areas involved in mating-induced pseudopregnancy. Because adrenal hormones are known to alter noradrenergic function, we examined whether adrenalectomy altered mating induced Fos expression in the A1 and A2 noradrenergic cell groups that project through the VNAB. Ovariectomized females were adrenalectomized (ADX) or sham operated (Sham) and, 2 weeks after surgery, were given oestrogen and progesterone and mated. They received 15 intromissions, five intromissions or 15 mounts without-intromission (mounts-only) from a male. Two hours after mating, rats were perfused and brains were collected; controls were perfused after being taken directly from their home cage. After immunocytochemical staining, Fos immunoreactive (Fos-IR) and dopamine-beta-hydroxylase-immunoreactive (DBH-IR) cells and the percentage of DBH cells that were labelled with Fos (% DBH/Fos) were counted. In the A1 area, Fos-IR and percentage DBH/Fos were not affected by adrenalectomy. Although an overall effect of mating treatment was found for both measures, no specific mating treatment increased labelled cells above home cage levels. In the caudal, middle and rostral A2, 15 intromissions induced a significant increase in Fos-IR in Sham females above all other groups and a higher percentage of DBH/Fos in the middle and rostral A2 areas. ADX females showed no rise in either Fos-IR or percentage DBH/Fos after 15 intromissions. However, in the middle and rostral A2, ADX females showed significantly increased Fos-IR and percentage DBH/Fos after mounts-only treatment above Sham mounts-only females and all other ADX groups. These results demonstrate that adrenal hormones suppress activation of A2 cells to mounts-only stimuli but contribute to A2 activation in response to intromissions from males. The latter effect may result from stress associated with receipt of vaginocervical stimulation during mating. PMID- 15344914 TI - Ageing and the diurnal expression of mRNAs for vasoactive intestinal peptide and for the VPAC2 and PAC1 receptors in the suprachiasmatic nucleus of male rats. AB - Ageing alters fundamental aspects of circadian rhythmicity in mammals; the effects include reduced rhythm amplitude and alterations in period length and in entrainment to the light/dark cycle. Such changes may reflect disruptions in cellular function within the suprachiasmatic nucleus (SCN), the site of the predominant circadian pacemaker. In the SCN, vasoactive intestinal peptide (VIP) synthesizing neurones receive various inputs, including retinohypothalamic projections containing pituitary adenylate cyclase activating peptide (PACAP). SCN VIP cells establish connections with local neurones and send efferents beyond the nucleus. Considerable evidence implicates VIP and PACAP in circadian rhythm maintenance and/or entrainment to photic Zeitgebers. These actions involve members of a distinct family of receptors; mRNAs for two such receptors, VPAC2 and PAC1, are present in the SCN. This study used isotopic in situ hybridization to examine the effects of ageing on expression of mRNAs for VIP, VPAC2 and PAC1 in the SCN of male rats under a 12 : 12 h light/dark cycle. Analysis of film autoradiographs from young adult (2-3 months) or aged (19-20 months) rats, at eight time points across the light/dark cycle, showed loss of diurnal rhythmicity and reduced levels for VIP mRNA in the aged group. A diurnal rhythm of VPAC2 receptor mRNA was present in both groups, but its levels were reduced in the aged rats. There were no differences between the two groups for PAC1 receptor mRNA expression. The present results indicate that ageing reduces VIP and VPAC2 receptor mRNA and eliminates diurnal expression of VIP mRNA within the SCN of aged male rats. PMID- 15344915 TI - Behavioural and neuroendocrine adaptations to repeated stress during puberty in male golden hamsters. AB - In adult animals, the consequences of stress are often severe and long lasting. Repeated subjugation in adult male golden hamsters inhibits aggression and increases submissive and avoidant behaviours. By contrast, subjugation during puberty enhances offensive aggression. The goals of this study were to characterize behavioural and neuroendocrine responses of naive and repeatedly subjugated juveniles to social defeat and to assess potential recovery from social stress. From the onset of puberty on postnatal day 28 (P28) to mid puberty (P42), animals were either socially subjugated or placed in a clean and empty cage for 20 min daily. The subjugated and control groups were further divided into subgroups and sacrificed under basal conditions or after social defeat on P28, P35 (early puberty), P45 (mid puberty) and P70 (early adulthood). On P35 and P45, repeatedly subjugated juveniles showed a complete inhibition of olfactory investigation (i.e. risk assessment) towards aggressive adults. Repeatedly subjugated also animals had lower postdefeat cortisol levels than controls on P45. Interestingly, basal cortisol levels increased gradually during puberty but did not differ between treatment groups at any point. Repeated subjugation was also associated with increased tyrosine hydroxylase immunoreactivity (ir-TH) within the extended medial amygdala. After a 4-week recovery period, none of these variables differed between subjugated and control groups. In an additional experiment, subjugated adults also had increased ir-TH in the medial extended amygdala, suggesting that these neurones are particularly responsive to social stress. In conclusion, puberty may be a developmental period characterized by behavioural and neuroendocrine plasticity in stress responsiveness. Furthermore, peri-pubertal changes in stress hormones may explain why juvenile hamsters are more resilient to social stress than adults. PMID- 15344917 TI - Neurosteroids: the StAR protein in the brain. AB - The nervous system is a well-known target for steroid hormones, as these hormones regulate brain functions that include neuronal survival and differentiation, myelination, neurogenesis, plasticity and repair after injury. Furthermore, the brain is also a steroidogenic tissue because it possesses the enzymes required to metabolize the common precursor, cholesterol, into steroids, named 'neurosteroids'. The rate-limiting step in the synthesis of steroid hormones is the access of cholesterol, accumulated in the outer mitochondrial membrane, to the first steroidogenic enzyme, P450scc (cholesterol side-chain cleavage enzyme), located in the inner mitochondrial membrane. In the ovary and the adrenal gland, which comprise classic steroidogenic tissues, this process requires the steroidogenic acute regulatory protein (StAR) to facilitate the shuttle of cholesterol through the intermembrane space. The mechanism used by the brain to regulate the first stage of steroidogenesis remains unknown. Recently, several groups have investigated the potential presence of StAR in the nervous tissue and have concluded that StAR is widely expressed throughout the brain, although restricted to specific cell populations. New results concerning localization, regulation and possible functions of StAR in the brain are discussed. PMID- 15344916 TI - 17Beta-oestradiol modulates glucocorticoid, neural and behavioural adaptations to repeated restraint stress in female rats. AB - Sex steroids have a role in modulating responses that extends beyond reproduction. The current study investigated the influence of the sex steroid 17beta-oestradiol on hypothalamic-pituitary-adrenal (HPA) and behavioural responses to acute or repeated restraint stress. Ovariectomized rats treated with 17beta-oestradiol or peanut oil via a subcutaneous silastic capsule were subjected to daily handling (non stressed), acute (single, 1 h) or daily (10 days, 1 h/day) restraint stress. Blood collected at the end of stress revealed that 17beta-oestradiol treatment augmented the corticosterone response to acute restraint. After daily exposure to restraint, the corticosterone response was noticeably diminished in untreated females but 17beta-oestradiol-treated rats still showed an exaggerated response compared to castrated, untreated females. Brain tissue collected 3 h after the end of restraint was probed using isotopic in situ hybridization for corticotropin-releasing factor (CRF) and vasopressin gene expression in the paraventricular nucleus (PVN) of the hypothalamus. 17beta oestradiol treatment at the higher dose (120 microg/ml) decreased basal CRF mRNA. Stress caused an increase in CRF mRNA expression in 17beta-oestradiol-treated rats but not in the vehicle group. Repeated restraint stress caused an increase in PVN parvocellular vasopressin gene expression, which was more pronounced in 17beta-oestradiol-replaced rats. Animals were exposed to the elevated plus maze for 5 min as a test for anxiety. Non-stressed control rats with or without 17beta oestradiol replacement spent the same percentage amount of time exploring the open arms of the maze. Previous exposure to acute restraint stress caused a marked reduction in the time spent exploring the open arms, indicating an increase in anxiety levels in these rats; this effect was observed in both vehicle and 17beta-oestradiol-treated rats. After repeated restraint stress, 17beta-oestradiol-replaced rats spent as much time exploring the open arms of the maze as controls, indicating adaptation. By contrast, nonreplaced rats were still showing a significant reduction in open arm exploration after repeated restraint. The present study presents novel data showing that the HPA axis remains reactive to repeated stress in 17beta-oestradiol-treated ovariectomized rats, but stress induced anxiety behaviour is reduced. PMID- 15344918 TI - Emerging functions of gonadotropin-releasing hormone II in mammalian physiology and behaviour. AB - Gonadotropin-releasing hormone (GnRH) is the central neuroendocrine regulator of the hypothalamic-pituitary-gonadal axis. Multiple structural variants of GnRH are present in vertebrates. The first isoform isolated in the mammalian brain (GnRH I) was shown to regulate the release of pituitary gonadotropins. Recently, a second form has been discovered in mammals (GnRH II), both in the brain and periphery. Although it is unlikely to be a primary regulator of gonadotropin release, the highly conserved GnRH II variant appears to have a wide array of physiological functions. In the periphery, GnRH I and II have similar roles in regulating cell proliferation and mediating hormonal secretion from the ovary and placenta in an autocrine/paracrine manner. In the brain, GnRH I and II apparently modulate mammalian reproductive behaviours in different but complementary ways: GnRH I stimulates luteinizing hormone/follicle-stimulating hormone secretion (and thus gonadal steroids) and promotes sexual behaviour in ad libitum fed animals. By contrast, GnRH II acts as a permissive regulator of female reproductive behaviour based on energy status, as well as a modifier of short-term food intake. GnRH II has also been implicated in the regulation of calcium and potassium channels in nervous systems of amphibians, functions which may also be present in mammals. Increasing evidence suggests that the effects of GnRH II in both the periphery and brain may be mediated by GnRH receptor subtypes distinct from the type-1 GnRH receptor. It is likely that this evolutionarily conserved peptide has been co-opted over evolutionary time to possess multiple regulatory functions in a broad range of biological aspects, including, but not limited to, reproduction. Here, the proposed actions of both neural and peripheral GnRH II in affecting physiology and behaviour are summarized, and an outline of critical directions for future research is proposed. PMID- 15344919 TI - Reduced aggression in AMPA-type glutamate receptor GluR-A subunit-deficient mice. AB - The importance of AMPA-type glutamate receptors has been demonstrated in neuronal plasticity and in adaptation to drugs of abuse. We studied the involvement of AMPA receptors in social interaction and anxiety and found that in several paradigms of agonistic behavior naive male mice deficient for the GluR-A subunit- containing AMPA receptors are less aggressive than wild-type littermates. GluR-A deficient mice and wild-type littermates exhibited similar basic behavior and reflexes as monitored by observational Irwin's test, but they tended to be less anxious in elevated plus-maze and light-dark tests. Maternal aggression or male female encounters were not affected which suggests that male hormones are involved in the expression of suppressed aggressiveness. However, testosterone levels and brain monoamines can be excluded and found to be similar between GluR A deficient and wild-type littermates. The reduced AMPA receptor levels caused by the lack of the GluR-A subunit, and measured by a 30% reduction in hippocampal [3H]-S-AMPA binding, seem to be the reason for suppressed male aggressiveness. When we analyzed mice with reduced number of functional AMPA receptors mediated by the genomic introduced GluR-A(Q582R) channel mutation, we observed again male specific suppressed aggression, providing additional evidence for GluR-A subunit containing AMPA receptor involvement in aggression. PMID- 15344920 TI - State-dependency in C. elegans. AB - Memory and the expression of learned behaviors by an organism are often triggered by contextual cues that resemble those that were present when the initial learning occurred. In state-dependent learning, the cue eliciting a learned behavior is a neuroactive drug; behaviors initially learned during exposure to centrally acting compounds such as ethanol are subsequently recalled better if the drug stimulus is again present during testing. Although state-dependent learning is well documented in many vertebrate systems, the molecular mechanisms underlying state-dependent learning and other forms of contextual learning are not understood. Here we demonstrate and present a genetic analysis of state- dependent adaptation in Caenorhabditis elegans. C. elegans normally exhibits adaptation, or reduced behavioral response, to an olfactory stimulus after prior exposure to the stimulus. If the adaptation to the olfactory stimulus is acquired during ethanol administration, the adaptation is subsequently displayed only if the ethanol stimulus is again present. cat-1 and cat-2 mutant animals are defective in dopaminergic neuron signaling and are impaired in state dependency, indicating that dopamine functions in state-dependent adaptation in C. elegans. PMID- 15344921 TI - Turning behavior in Drosophila larvae: a role for the small scribbler transcript. AB - The Drosophila larva is extensively used for studies of neural development and function, yet the mechanisms underlying the appropriate development of its stereotypic motor behaviors remain largely unknown. We have previously shown that mutations in scribbler (sbb), a gene encoding two transcripts widely expressed in the nervous system, cause abnormally frequent episodes of turning in the third instar larva. Here we report that hypomorphic sbb mutant larvae display aberrant turning from the second instar stage onwards. We focus on the smaller of the two sbb transcripts and show that its pan-neural expression during early larval life, but not in later larval life, restores wild type turning behavior. To identify the classes of neurons in which this sbb transcript is involved, we carried out transgenic rescue experiments. Targeted expression of the small sbb transcript using the cha-GAL4 driver was sufficient to restore wild type turning behavior. In contrast, expression of this sbb transcript in motoneurons, sensory neurons or large numbers of unidentified interneurons was not sufficient. Our data suggest that the expression of the smaller sbb transcript may be needed in a subset of neurons for the maintenance of normal turning behavior in Drosophila larvae. PMID- 15344922 TI - Sociability and preference for social novelty in five inbred strains: an approach to assess autistic-like behavior in mice. AB - Deficits in social interaction are important early markers for autism and related neurodevelopmental disorders with strong genetic components. Standardized behavioral assays that measure the preference of mice for initiating social interactions with novel conspecifics would be of great value for mutant mouse models of autism. We developed a new procedure to assess sociability and the preference for social novelty in mice. To quantitate sociability, each mouse was scored on measures of exploration in a central habituated area, a side chamber containing an unfamiliar conspecific (stranger 1) in a wire cage, or an empty side chamber. In a secondary test, preference for social novelty was quantitated by presenting the test mouse with a choice between the first, now-familiar, conspecific (stranger 1) in one side chamber, and a second unfamiliar mouse (stranger 2) in the other side chamber. Parameters scored included time spent in each chamber and number of entries into the chambers. Five inbred strains of mice were tested, C57BL/6J, DBA/2J, FVB/NJ, A/J and B6129PF2/J hybrids. Four strains showed significant levels of sociability (spend- ing more time in the chamber containing stranger 1 than in the empty chamber) and a preference for social novelty (spending more time in the chamber containing stranger 2 than in the chamber containing the now-familiar stranger 1). These social preferences were observed in both male and female mice, and in juveniles and adults. The exception was A/J, a strain that demonstrated a preference for the central chamber. Results are discussed in terms of potential applications of the new methods, and the proper controls for the interpretation of social behavior data, including assays for health, relevant sensory abilities and motor functions. This new standardized procedure to quantitate sociability and preference for social novelty in mice provides a method to assess tendencies for social avoidance in mouse models of autism. PMID- 15344923 TI - Automated apparatus for quantitation of social approach behaviors in mice. AB - Mouse models of social dysfunction, designed to investigate the complex genetics of social behaviors, require an objective methodology for scoring social interactions relevant to human disease symptoms. Here we describe an automated, three chambered apparatus designed to monitor social interaction in the mouse. Time spent in each chamber and the number of entries are scored automatically by a system detecting photocell beam breaks. When tested with the automated equipment, juvenile male C57BL/6J mice spent more time in a chamber containing a stranger mouse than in an empty chamber (sociability), similar to results obtained by the observer scored method. In addition, automated scoring detected a preference to spend more time with an unfamiliar stranger than a more familiar conspecific (preference for social novelty), similar to results obtained by the observer scored method. Sniffing directed at the wire cage containing the stranger mouse correlated significantly with time spent in that chamber, indicating that duration in a chamber represents true social approach behavior. Number of entries between chambers did not correlate with duration of time spent in the chambers; entries instead proved a useful control measure of general activity. The most significant social approach behavior took place in the first five minutes of both the sociability and preference for social novelty tests. Application of these methods to C57BL/6J, DBA/2J and FVB/NJ adult males revealed that all three strains displayed tendencies for sociability and preference for social novelty. To evaluate the importance of the strain of the stranger mouse on sociability and preference for social novelty, C57BL/6J subject mice were tested either with A/J strangers or with C57BL/6J strangers. Sociability and preference for social novelty were similar with both stranger strains. The automated equipment provides an accurate and objective approach to measuring social tendencies in mice. Its use may allow higher-throughput scoring of mouse social behaviors in mouse models of social dysfunction. PMID- 15344925 TI - The bugs that came in from the cold. PMID- 15344926 TI - Illuminating the detection chain of bacterial bioreporters. AB - Engineering bacteria for measuring chemicals of environmental or toxicological concern (bioreporter bacteria) has grown slowly into a mature research area. Despite many potential advantages, current bioreporters do not perform well enough to comply with environmental detection standards. Basically, the reasons for this are the lack of engineering principles in the detection chain in the bioreporters. Here, we dissect critical steps in the detection chain and illustrate how bioreporter design could be improved by mutagenizing specificity and selectivity of the sensing and regulatory proteins, by newer expression strategies and application of different signalling networks. Furthermore, we describe how redesigning bioreporter assays with respect to pollutant transport into the cells and application of other detection devices can decrease detection limits and increase the speed of detection. PMID- 15344927 TI - Plasmolysis induced by toluene in a cyoB mutant of Pseudomonas putida. AB - The cyoABCDE gene cluster of Pseudomonas putida DOT-T1E encodes a terminal cytochrome oxidase. A 500-bp 'cyoB' DNA fragment was cloned in pCHESI Omega Km and used to generate a cyoB knock-out mutant in vivo. The mutant strain was not limited in the generation of proton-motif force, although when grown on minimal medium with glucose or citrate, the CyoB mutant exhibited a slight increase in duplication time with respect to the wild-type strain. This effect was even more pronounced when toluene was supplied in the gas phase. In consonance with the negative effect of toluene on the growth was the finding that the CyoB mutant was hypersensitive to sudden 0.3% (v/v) toluene shocks, in contrast with the wild type strain. This effect was particularly exacerbated in cells that reached the stationary phase. The increased sensitivity to solvents of the CyoB mutant did not appear to be related to the inability of the cells to strengthen the membrane package or to induce the efflux pumps in response to the solvent, but rather to solvent-induced plasmolysis that may be triggered by wrinkles in the cytoplasmic membrane at the poles of the mutant cells, and invagination of the outer membranes, which eventually lead to cell death. PMID- 15344928 TI - Diversity of Frankia strains associated to Myrica gale in Western Europe: impact of host plant (Myrica vs. Alnus) and of edaphic factors. AB - Myricaceae can be nodulated by a variety of Frankia strains isolated from other actinorhizal families. Consequently, the genus Myrica has been considered to have low specificity with respect to microsymbiont taxa. In contrast to controlled studies of Myrica infectious capacity, field studies in North America have indicated that M. gale symbionts belong to the genetic group of Alnus-infective strains. Myrica gale is the most widely distributed species in the genus so this study focused on describing the genetic diversity of M. gale-nodulating strains from 10 sites in Western Europe across a range of edaphic conditions. When possible, the specificity of M. gale-infective strains was compared with that of Alnus-infective strains from the same sites. Nodular strains from Belgium, France and Spain were characterized using PCR-RFLP of rrs gene and 16S-23S IGS. rrs-RFLP patterns showed a high level of homogeneity among European strains with one dominant genotype. IGS-RFLP patterns revealed the largest inter and intrasite diversity in France. In Belgium, Frankia strains were found to occur in two groups according to soil pH and organic matter characteristics of the sites. European M. gale-infective strains were genetically different from European Alnus and North American M. gale-infective strains indicating the possibility of different pathways of co-evolution among geographically isolated populations. PMID- 15344929 TI - Intracellular manganese granules formed by a subsurface bacterium. AB - The demonstrated ability of prokaryotes to form internal metal oxide particles during active metabolism has been restricted to Fe. Mineral-bound Mn(IV) is a known electron acceptor during dissimilatory metal reduction by Shewanella putrefaciens, yet no internal deposits of Mn have been reported to form during anaerobic respiration. We observed distinct nanometre-sized Mn-rich granules in the cytoplasm when either birnessite or pyrolusite (beta-MnO(2)) served as the electron acceptor during growth. During rapid Mn reduction, additional precipitates of Mn were also observed in the periplasm together with the cytoplasmic granules. The bacteria did not accumulate detectable Mn in the outer membrane during formation of the internal precipitates. This is the first report of an intracellular Mn solid produced by bacteria and coupled anaerobically to DR. PMID- 15344930 TI - Molecular diversity of methanotrophs in Transbaikal soda lake sediments and identification of potentially active populations by stable isotope probing. AB - Soda lakes are an environment with an unusually high pH and often high salinity. To identify the active methanotrophs in the Soda lake sediments, sediment slurries were incubated with a 10% (v/v) (13)CH(4) headspace and the (13)C labelled DNA was subsequently extracted from these sediments following CsCl density gradient centrifugation. This DNA was then used as a template for PCR amplification of 16S rRNA genes and genes encoding PmoA and MmoX of methane monooxygenase, key enzymes in the methane oxidation pathway. Phylogenetic analysis of 16S rRNA genes, PmoA and MmoX identified that strains of Methylomicrobium, Methylobacter, Methylomonas and 'Methylothermus' had assimilated the (13)CH(4). Phylogenetic analysis of PmoA sequences amplified from DNA extracted from Soda lake sediments before Stable Isotope Probing (SIP) treatment showed that a much wider diversity of both type I and type II methanotroph sequences are present in this alkaline environment. The majority of methanotroph sequences detected in the (13)C-DNA studies were from type I methanotrophs, with 50% of 16S rRNA clones and 100% of pmoA clones from both Lake Suduntuiskii Torom and Lake Gorbunka suggesting that the type I methanotrophs are probably responsible for the majority of methane oxidation in this environment. PMID- 15344931 TI - Quantifying 3H-thymidine incorporation rates by a phylogenetically defined group of marine planktonic bacteria (Bacteriodetes phylum). AB - The rate of [(3)H-methyl] thymidine ((3)H-TdR) incorporation into DNA has been applied extensively to measure cell production by bacterial communities in aquatic environments. Here we describe a method to quantify (3)H-TdR incorporation by specific, phylogenetically defined members of the bacterial community. The method involves selectively capturing DNA from targeted groups of bacteria and then quantifying its (3)H radioactivity. The method was applied to measure (3)H-TdR incorporation by the members of the phylum Bacteriodetes whose members, which include the Cytophaga-Flavobacter cluster, are ubiquitous in coastal waters. (3)H-labelled DNA from Bacteriodetes was selectively biotinylated in PCR-like reactions that contained a Bacteriodetes-specific 16S rRNA gene primer, thermostable DNA polymerase and biotinylated dUTP. The biotinylated DNA was then captured on streptavidin-coated beads and its (3)H radioactivity determined by scintillation counting. We have termed this method 'selective nucleic acid polymerase-biotinylation and capture' or 'SNAP-BAC'. Internal (33)P labelled DNA standards were used to quantify the recovery of (3)H-labelled DNA from the SNAP-BAC reactions. The method was verified by successfully targeting Bacteriodetes in simple laboratory mixtures of (3)H-labelled DNA extracted from pure cultures of Bacteriodetes and gamma-proteobacteria. Field application of this method in Puget Sound and off the Washington coast determined that Bacteriodetes were responsible for 56 +/- 17% and 32 +/- 5% of community (3)H-TdR incorporation (1.3 +/- 0.3 and 9.9 +/- 1.7 pmol l(-1) h(-1)) at these two locations. PMID- 15344932 TI - Impact of lime, nitrogen and plant species on bacterial community structure in grassland microcosms. AB - A microcosm-based approach was used to study impacts of plant and chemical factors on the bacterial community structure of an upland acidic grassland soil. Seven perennial plant species typical of both natural, unimproved (Nardus stricta, Agrostis capillaris, Festuca ovina and F. rubra) and fertilized, improved (Holcus lanatus, Lolium perenne and Trifolium repens) grasslands were either left unamended or treated with lime, nitrogen, or lime plus nitrogen in a 75-day glasshouse experiment. Lime and nitrogen amendment were shown to have a greater effect on microbial activity, biomass and bacterial ribotype number than plant species. Liming increased soil pH, microbial activity and biomass, while decreasing ribotype number. Nitrogen addition decreased soil pH, microbial activity and ribotype number. Addition of lime plus nitrogen had intermediate effects, which appeared to be driven more by lime than nitrogen. Terminal restriction fragment length polymorphism (TRFLP) analysis revealed that lime and nitrogen addition altered soil bacterial community structure, while plant species had little effect. These results were further confirmed by multivariate redundancy analysis, and suggest that soil lime and nitrogen status are more important controllers of bacterial community structure than plant rhizosphere effects. PMID- 15344933 TI - Estimating bacterial diversity from clone libraries with flat rank abundance distributions. AB - There are a number of parametric and non-parametric methods for estimating diversity. However all such methods employ either the proportional abundance of the most abundant taxon in a sample or require that a specific taxon is sampled more than once. Consequently, the available methods for estimating diversity cannot be applied to samples consisting entirely of singletons, which might be characteristic of some hyperdiverse communities. Here we present a non-parametric method that estimates the probability that a given number of unique taxa would be sampled from a community with a particular diversity. We have applied this approach to a well known data set of 100 unique clones from a sample of Amazonian soil (Borneman and Triplett (1997) Appl Environ Microbiol 63: 2647-2653) and determine the probability that this observation would be made from an environment of a given diversity. On this basis we can state this observation would be very unlikely (P = 0.006) if the soil diversity was less than 10(3), and quite unlikely (P = 0.6) if the diversity was less than 10(4), and probable (P = 0.95) if the diversity was about 10(5). There are essentially no contestable assumptions in our method. Thus we are able to offer almost unequivocal evidence that the bacterial diversity, of at least soils, is very large and a method that may be used to interpret samples consisting entirely of singletons from other hyperdiverse communities. PMID- 15344934 TI - Low archaeal diversity linked to subseafloor geochemical processes at the Lost City Hydrothermal Field, Mid-Atlantic Ridge. AB - The recently discovered Lost City Hydrothermal Field (LCHF) represents a new type of submarine hydrothermal system driven primarily by exothermic serpentinization reactions in ultramafic oceanic crust. Highly reducing, alkaline hydrothermal environments at the LCHF produce considerable quantities of hydrogen, methane and organic molecules through chemo- and biosynthetic reactions. Here, we report the first analyses of microbial communities inhabiting carbonate chimneys awash in warm, high pH fluids at the LCHF and the predominance of a single group of methane-metabolizing Archaea. The predominant phylotype, related to the Methanosarcinales, formed tens of micrometre-thick biofilms in regions adjacent to hydrothermal flow. Exterior portions of active structures harboured a diverse microbial community composed primarily of filamentous Eubacteria that resembled sulphide-oxidizing species. Inactive samples, away from regions of hydrothermal flow, contained phylotypes related to pelagic microorganisms. The abundance of organisms linked to the volatile chemistry at the LCHF hints that similar metabolic processes may operate in the subseafloor. These results expand the range of known geological settings that support biological activity to include submarine hydrothermal systems that are not dependent upon magmatic heat sources. PMID- 15344935 TI - Microbial diversity of extant stromatolites in the hypersaline marine environment of Shark Bay, Australia. AB - Stromatolites have been present on Earth, at various levels of distribution and diversity, for more than 3 billion years. Today, the best examples of stromatolites forming in hypersaline marine environments are in Hamelin Pool at Shark Bay, Western Australia. Despite their evolutionary significance, little is known about their associated microbial communities. Using a polyphasic approach of culture-dependent and culture-independent methods, we report the discovery of a wide range of microorganisms associated with these biosedimentary structures. There are no comparable reports combining these methodologies in the survey of cyanobacteria, bacteria, and archaea in marine stromatolites. The community was characterized by organisms of the cyanobacterial genera Synechococcus, Xenococcus, Microcoleus, Leptolyngbya, Plectonema, Symploca, Cyanothece, Pleurocapsa and Nostoc. We also report the discovery of potentially free-living Prochloron. The other eubacterial isolates and clones clustered into seven phylogenetic groups: OP9, OP10, Marine A group, Proteobacteria, Low G+C Gram positive, Planctomycetes and Acidobacteria. We also demonstrate the presence of sequences corresponding to members of halophilic archaea of the divisions Euryarchaeota and Crenarchaeota and methanogenic archaea of the order Methanosarcinales. This is the first report of such archaeal diversity from this environment. This study provides a better understanding of the microbial community associated with these living rocks. PMID- 15344936 TI - Molecular evidence that plastids in the toxin-producing dinoflagellate genus Dinophysis originate from the free-living cryptophyte Teleaulax amphioxeia. AB - Some species of the dinoflagellate genus Dinophysis form red tides and are toxin producers with a great environmental impact. The dinoflagellates as a group display high plastid diversity. Several cases indicate that plastids have been replaced. In the case of the genus Dinophysis, the plastids show characteristics of a plastid originating from a cryptophyte. Recent molecular evidence showed that the plastid indeed originates from a cryptophyte, but the source could not be identified to species or genus level. The data presented here show that both a 799 bp region of the psbA gene and 1,221 bp region of the 16S rRNA gene from Dinophysis spp. are identical to the same loci in Teleaulax amphioxeia SCCAP K434. This strongly indicates that the plastid was acquired recently in Dinophysis and may be a so-called kleptoplastid, specifically originating from a species of Teleaulax. PMID- 15344938 TI - Editorial: Reproductive health and conflict: looking back and moving ahead. PMID- 15344939 TI - Changes in HIV/AIDS/STI knowledge, attitudes and practices among commercial sex workers and military forces in Port Loko, Sierra Leone. AB - Sierra Leone suffered from 11 years of civil war (1991-2002) resulting in tens of thousands of deaths and mutilations together with massive population displacement. In 2001, ARC International, Sierra Leone conducted a baseline survey of 201 commercial sex workers (CSWs) and 202 military respondents on the knowledge, attitudes and practices surrounding HIV/AIDS and STIs in Port Loko, Sierra Leone. In 2003, a comparable post-intervention survey of 202 CSWs and 205 military respondents was performed. Comparison of baseline and post-intervention results showed that HIV/AIDS knowledge increased among both groups, with those able to name three effective means of avoiding AIDS increasing from 5 per cent to 70 per cent among CSWs, and 11 to 75 per cent among the military. Reported condom use during last sex increased among CSWs from 38 to 68 per cent and among military from 39 to 68 per cent. These results demonstrate that, despite the challenges inherent in a post-conflict country, good-quality AIDS-prevention programmes can be effective. PMID- 15344940 TI - Using evidence to improve reproductive health quality along the Thailand-Burma border. AB - The Mae Tao Clinic, located on the Thailand-Burma border, has provided health services for illegal migrant workers in Thailand and internally displaced people from Burma since 1989. In 2001, the clinic launched a project with the primary aim of improving reproductive health services and the secondary aim of building clinic capacity in monitoring and evaluation (M&E). This paper first presents the project's methods and key results. The team used observation of antenatal care and family-planning sessions and client exit interviews at baseline and follow up, approximately 13 months apart, to assess performance on six elements of quality of care. Findings indicated that improving programme readiness contributed to some improvement in the quality of services, though inconsistencies in findings across the methods require further research. The paper then identifies lessons learned from introducing M&E in a resource constrained setting. One key lesson was that a participatory approach to M&E increased people's feelings of ownership of the project and motivated staff to collect and use data for programme decision-making to improve quality. PMID- 15344941 TI - Reproductive health in Afghanistan: results of a knowledge, attitudes and practices survey among Afghan women in Kabul. AB - A reproductive-health knowledge, attitudes and practices (KAP) survey was carried out among 468 Afghan women of reproductive age. A convenience sample of women was selected from attendees in the outpatient departments of four health facilities in Kabul. Seventy-nine per cent of respondents had attended at least one antenatal consultation during their last pregnancy. Two-thirds (67 per cent) delivered their first child between 13 and 19 years. The Caesarean-section rate was low (1.6 per cent). Two-thirds (67 per cent) of deliveries occurred in the home. The contraceptive prevalence rate was 23 per cent (16 per cent modern and 7 per cent natural methods). Twenty-four per cent had knowledge of any STIs, although most of these women did not know correctly how to prevent them. Most of the women (93 per cent) needed authorization from their husband or a male relative before seeking professional health-care. In multivariate analysis, women's schooling was significantly associated with antenatal-care attendance (AOR 4.78), institutional delivery (AOR 2.29), skilled attendance at birth (AOR 2.07) and use of family planning (AOR 4.59). Reproductive-health indicators were noted to be poor even among these women living in Kabul, a group often considered to be the most privileged. To meet the reproductive-health needs of Afghan women, the socio-cultural aspects of their situation--especially their decision-making abilities -- will need to be addressed. A long-standing commitment from agencies and donors is required, in which the education of women should be placed as a cornerstone of the reconstruction process of Afghanistan. PMID- 15344942 TI - Effects of social interaction between Afghan refugees and Iranians on reproductive health attitudes. AB - Afghan refugees in Iran, like many Muslims around the world, think that using contraceptives is against their religious beliefs. The majority of Iranians also thought so until a decade ago. Since then an all-encompassing social movement has emerged in Iran in which women and young people have played decisive roles. This movement has led to an attitude shift towards acceptance of family planning across Iranian society. High-ranking clerics, responding to the social movement, issued progressive edicts indicating that poor families could practise family planning. This paper reports the findings of focus group studies carried out among Afghan refugee men in two communities in Iran. The findings indicate that exposure to Iranian life, especially the rulings of Iranian clerics, have influenced Afghans' views on family planning. The findings also indicate that a system of justification is needed to help people avoid both social sanctions and individual stresses resulting from a controversial practice regarded as a sin for centuries. PMID- 15344943 TI - A determination of the prevalence of gender-based violence among conflict affected populations in East Timor. AB - The Reproductive Health Response in Conflict (RHRC) Consortium designed a standardised questionnaire to measure gender-based violence (GBV) prevalence in conflict-affected settings. A preliminary field test was undertaken July-August 2002 in one urban and one rural district in East Timor to assess the prevalence of GBV among women 18-49 years of age during and after conflict. The field test used a cross-sectional survey design with a two-stage random selection process. During the year preceding East Timor's 1999 crisis, 23.8 per cent of respondents reported physical assault by an intimate partner; this rate was not significantly different in the year preceding the survey (24.8 per cent). Assault by perpetrators outside the family declined significantly from 24.2 per cent during the crisis to 5.8 per cent post-crisis for physical assault (p<.001) and 22.7 per cent during the crisis to 9.7 per cent post-crisis for sexual assault (p=0.046). The field test stimulated and informed additional research in East Timor, and the complementary findings of these research initiatives continue to be used to develop local policies and programming to prevent and address GBV. PMID- 15344944 TI - HIV/AIDS among conflict-affected and displaced populations: dispelling myths and taking action. AB - Conflict, displacement, food insecurity and poverty make affected populations more vulnerable to HIV transmission. However, the common assumption that this vulnerability necessarily translates into more HIV infections and consequently fuels the HIV/AIDS epidemic is not supported by data. Whether or not conflict and displacement affect HIV transmission depends upon numerous competing and interacting factors. This paper explores and explains the epidemiology of HIV/AIDS in conflict and addresses the unique characteristics that must be addressed when planning and implementing HIV/AIDS interventions among populations affected by conflict as compared with those in resource-poor settings. These include targeting at-risk groups, protection, programming strategies, coordination and integration and monitoring and evaluation. Areas for future HIV/AIDS operational research in conflict are discussed. PMID- 15344945 TI - Thinking ahead about reproductive health: contingency planning and emergency preparedness in crisis situations (Iraq and West Africa). AB - The United Nations Population Fund (UNFPA) coordinated efforts to integrate RH into contingency planning for the 2003 Iraq crisis and the 2003 regional response for displaced populations in West Africa. Using UNFPA's network of country offices in the Middle East, staff developed logistics plans, conducted workshops and pre-positioned RH supplies. Though refugee movements did not occur, the contingency planning enhanced the response capacity of UNFPA offices and made it possible to rapidly provide assistance inside Iraq. In West Africa, multi-country workshops and follow-up resulted in country-level and regional action plans useful during the renewed crises of 2003; scarce funding, however, limited their full implementation. UNFPA's experiences show that contingency planning requires committing resources for crises, some of which will not occur; new staff skills; and follow-up. Moreover, RH is considered by some to be additional to the core elements of contingency planning. RH's political sensitivity, particularly with certain donors, further complicated integrated planning. PMID- 15344946 TI - Abstracts of the 43rd Annual Scientific Meeting of the British Society for Clinical Cytology. September 12-15, 2004. Sheffield, United Kingdom. PMID- 15344948 TI - Mycophenolates in transplantation. AB - The discovery of mycophenolic acid (MPA) as a potent immunosuppressant, able to inhibit B- and T-cell proliferation by blocking production of guanosine nucleotides required for DNA synthesis, allowed its potential in the field of transplantation to be realized. Mycophenolate mofetil (MMF), an MPA prodrug, has been shown to be an effective immunosuppressant in transplant therapy. Clinical trials in renal, heart, and liver transplant recipients have demonstrated that, in combination with cyclosporine and steroids, MMF therapy can reduce the incidence and severity of acute rejection episodes and improve graft and patient survival as well as graft function. Although MMF is generally well tolerated, optimal therapy may be limited by associated side effects, in particular gastrointestinal (GI) toxicity, which may occur in over 40% of patients. Dose changes resulting from GI side effects may potentially lead to sub-therapeutic dosing and impaired clinical outcomes. An enteric-coated formulation delivering MPA - enteric-coated mycophenolate sodium (EC-MPS) has been developed to improve MPA-related upper GI adverse events. EC-MPS delays the release of MPA, consistent with a functional enteric-coating. Recent clinical trials have demonstrated that EC-MPS is as effective and safe as MMF in both de novo and maintenance renal transplant patients. PMID- 15344949 TI - Early experience with two-dose daclizumab in the prevention of acute rejection in cardiac transplantation. AB - BACKGROUND: Daclizumab is a human monoclonal antibody that binds to the interleukin-2 receptor. It has been used as induction therapy in heart transplantation with repeated administrations over several weeks. At our institution, we use a two-dose regimen of daclizumab based on its extended half life. We sought to determine the incidence of acute rejection with 2-dose daclizumab in cardiac transplantation. METHODS: Eighteen consecutive heart transplants performed at a single center were analyzed retrospectively. Patients received daclizumab (2 mg/kg) within 8 h of cardiac transplantation and a second dose (1 mg/kg) 2 wk thereafter. Maintenance immunosupression included mycophenolate mofetil, prednisone and either cyclosporine or tacrolimus, based on side-effect profile. The endpoint was the incidence of acute rejection as defined by a histologic grade >2 according to the classification of the International Society of Heart and Lung Transplantation. RESULTS: Four patients had acute rejections (all were 3A) during the first 3 months post-transplantation. All four patients had rejection at the first biopsy and only two had rejection thereafter. None of the rejections were hemodynamically significant and no patients were hospitalized. All except one rejection was seen in the context of low 2-h cyclosporine levels. The two-dose regimen was easier to administer on an outpatient basis and resulted in lower cost. CONCLUSIONS: This preliminary report suggests that induction therapy with a two-dose regimen of daclizumab appears to be safe and well tolerated in patients undergoing cardiac transplantation. PMID- 15344950 TI - Optimal timing for living-related liver transplantation in children. AB - BACKGROUND: The pediatric end stage liver disease (PELD) score has been used widely to prioritize children awaiting cadaveric liver transplantation (LTx). To establish the objective parameter for optimal timing of living-related LTx (LRLTx), we have assessed our cases using the PELD score. METHODS: From 1997 to 2002, 24 children were evaluated 28 times for the indication of LRLTx. Among them, 15 were for jaundice and nine for growth failure, hepatopulmonary syndrome, and variceal bleeding. Nine of 24 children underwent LRLTx. They were divided into several groups according to their clinical course. The PELD score consisted of age, albumin, total bilirubin, prothrombine time-international ratio (INR) and growth failure. A cut-off value was obtained by the highest positive and negative predictive value. RESULTS: The PELD score in cases whose indication for LRLTx was approved was significantly higher compared with the cases who were not, and a cut off value of 4 was obtained. The PELD score in cases who were alive after LRLTx was significantly lower compared with the cases who died after LRLTx or evaluation of the indication, and a cut-off value of 22 was established. CONCLUSION: LRLTx may be considered when the PELD score exceeds 4, and LRLTx may be required immediately when the PELD score exceeds 22. PMID- 15344951 TI - Non-resective ablation therapy for hepatocellular carcinoma: effectiveness measured by intention-to-treat and dropout from liver transplant waiting list. AB - BACKGROUND: Orthotopic liver transplantation (OLT) for patients with small hepatocellular carcinoma (HCC) is widely accepted, and the usefulness of local ablation techniques as a bridge for liver transplantation is still under investigation. METHODS: From December 1997 to February 2003, patients with cirrhosis and T0-T1-T2-T3 stage HCC received multi-modality ablative therapy (MMT) for the treatment of their HCC and were evaluated for OLT; listed, and transplanted when an allograft became available. MMT included radiofrequency ablation (RFA), and/or Trans-Arterial Chemo-Embolization (TACE), and alcohol (EtOH) ablation, followed by Trans-Arterial Chemo-Infusion (TACI), with repeated treatments based on follow up hepatic magnetic resonance imaging (MRI) during the waiting period for OLT. RESULTS: A total of 135 HCC patients were seen at our center within this time frame. The intention-to-treat group included 33 (24.4%) patients with T0, T1, T2, T3 HCC and cirrhosis. There were 31 men and two women. The mean age was 53.6 +/- 7.2 yr. All patients received MMT with a mean of 2.90 +/- 1.5 procedures per patient. Tumor-node-metastasis (TNM) stages at time of listing were: T0 in one patient, T1 in nine patients, T2 in 17 patients, and T3 in six patients. Twenty-eight (85%) patients have received OLT. Five (12.19%) patients were listed and removed (dropout) from the transplant waiting list after waiting 5, 5, 5, 8, and 14 months respectively. The waiting time of the HCC listed group was 9.1 +/- 14.8 months with a mean follow up of 32 months. OLT patient survival and cancer-free survival are 92.9% and 95.24%, respectively; the overall survival of intention-to-treat group was 79% at 32 months follow up. Predictors of dropout included an alpha-fetoprotein (AFP, >400 ng/mL) and T3 HCC stage. CONCLUSION: Aggressive ablation therapy with a short transplant waiting time optimizes the use of OLT for curative intent in selective cirrhotic HCC patients. PMID- 15344952 TI - Detection of cardiac allograft rejection by real-time PCR analysis of circulating mononuclear cells. AB - BACKGROUND: Detection of cardiac allograft rejection is based on the histological examination of endomyocardial biopsies (EMB). We have explored the possibility of whether graft rejection could be detected by characteristic gene expression patterns in peripheral blood mononuclear cells (PBMC) of heart-transplant recipients. METHODS: The study included 58 blood samples of 44 patients. On the day of EMB, mononuclear cells were isolated from peripheral blood, and gene expression was measured by quantitative real-time PCR. Thirty-nine parameters, including cytokine and chemokine genes were analyzed. Gene expression results were correlated with histological assessment of concomitant evaluated EMB according to International Society for Heart and Lung Transplantation (ISHLT) nomenclature. RESULTS: Gene expression of perforin, CD95 ligand, granzyme B, RANTES, CXCR3, COX2, ENA 78 and TGF-beta1 was significantly different in PBMC of patients with mild to moderate degrees of allograft rejection (> or =grade 2) compared with patients exhibiting no or minor forms of rejection ( or =grade 2 vs. or =60 yr', 'waiting for transplant > or =6 yr', and 'human leukocyte antigen DR (HLA-DR) mismatch', based on the multivariate model. These factors may reflect the scarcity of organ donations in Israel, which leads to a prolonged waiting time, higher tolerance for HLA-DR mismatches, and utilization of kidneys from elderly donors. PMID- 15344963 TI - Long-term follow-up of pediatric transplant recipients: mycophenolic acid trough levels are not a good indicator for long-term graft function. AB - BACKGROUND: Mycophenolate mofetil (MMF) has the potential of decreasing acute rejection episodes early following renal transplantation. Pharmocokinetic monitoring of mycophenolic acid (MPA) trough levels is performed by many centers. MMF has also proved successful in improving long-term graft function in patients with chronic allograft nephropathy (CAN). However, no data for long-term monitoring of MPA in children have yet been published. METHODS: MMF therapy with a dose of 600 mg/m2 twice daily was initiated in 42 children (median age 9.4 yr, range 1.4-15.1) after a median period of 3.8 yr (range 1.0-10.6) post transplantation-- according to significant increases in serum creatinine. CAN was diagnosed by renal biopsy and the amount of fibrosis was quantified with PicroSiriusRed staining. MMF therapy was combined with ciclosporin A and prednisolone. MPA-C0-levels, measured by high-pressure liquid chromatography, were tested every 3 months. In 12 children a full MPA area under the curve concentration (AUC) was measured. The glomerular filtration rate (GFR) was calculated at the start of MMF therapy and 2 yr later. RESULTS: After initiation of MMF, the calculated GFR did not decrease further in 22 children and mean GFR remained stable for 2 yr in the whole study group. There was a significant correlation between MPA levels 75 min after administration and the full AUC (r = 0.94, p < 0.001) but no correlation between trough levels and AUC (r = -0.07, p > 0.05). The mean MPA trough level was 2.8 +/- 1.3 ng/mL. The intra-individual coefficient of variation was 2.6 +/- 1.4. There was no correlation between mean MPA trough levels and GFR development after 2 yr (r = 0.03, p > 0.05). In children with an MPA level below 1.2 mg/L (n = 5), the mean GFR decline was no different to those with a higher level (p > 0.05). CONCLUSIONS: Drug monitoring of MPA trough levels had no impact on long-term graft function in kidney recipients. MPA levels taken 75 min after administration showed a high correlation with MPA-AUC whereas C0-levels did not correlate. The value of C75 drug measurements for monitoring renal allograft survival will have to be judged in future studies. PMID- 15344964 TI - Endoscopic diagnosis of cytomegalovirus infection of upper gastrointestinal tract in solid organ transplant recipients: Hungarian single-center experience. AB - BACKGROUND: Cytomegalovirus (CMV) is considered to be the major cause of upper gastrointestinal (GI) symptoms in organ transplant recipients. In the diagnosis of GI CMV infection the detection of the virus in the mucosa is essential. The aim of the study was to evaluate the significance of CMV, detected in biopsy specimens from stomach and duodenum of solid organ transplant recipients. METHODS: Data of 227 elective upper endoscopies on symptomatic organ transplant recipients were evaluated for clinical symptoms, endoscopic changes and conventional histologic alterations of mucosal biopsy samples. Qualitative PCR was performed for detection of the presence of CMV-DNA in each biopsy materials. RESULTS: CMV-DNA was detected in biopsy samples of 91 patients (40.1%) while only in 20 cases (8.8%) the signs of CMV infections were found by conventional histology (p < 0.00001). No considerable differences could be observed in symptomatic, histologic alterations between CMV-PCR positive and negative groups. There were no endoscopic changes in 25.3% of CMV-PCR positive and 5.1% of negative patients. CONCLUSIONS: Qualitative PCR is an accurate method for the detection of CMV in the mucosa of the GI tract. Further investigations are needed for determination of the exact pathological role of detected CMV. PMID- 15344965 TI - Severe elevations of FK506 blood concentration due to diarrhea in renal transplant recipients. AB - BACKGROUND: The rate of metabolism in the intestine of oral administered FK506 decreases as FK506 passes on to the lower intestine. In transplant recipients with diarrhea given oral FK506, the main areas for absorption of FK506 shift to the lower intestine, where the ability to metabolize FK506 is weaker. Therefore it is considered likely that when FK506 is administered to recipients with diarrhea, the blood concentration of FK506 will be higher. MATERIAL AND METHODS: Twenty recipients experiencing episodes of diarrhea were investigated to determine the trough level of FK506 and the time required for the FK506 trough level to return to the level that obtained before diarrhea. AUC0-4h and Cmax of FK506 were investigated in eight recipients. In cases with severe diarrhea, the daily fluctuations of FK506 blood concentration were also investigated. RESULT: The FK506 trough level (p < 0.0001), AUC (p = 0.0173), and Cmax (p = 0.0173) were found to be significantly higher during episodes of diarrhea. In almost all cases, it took between 2 and 4 wk for the elevated FK506 trough level to return to its previous level following a bout of diarrhea. In the daily fluctuations of FK506 concentration, Tmax was prolonged. In some cases, the concentration was highest just before administration of FK506, when it should have been at trough level. CONCLUSIONS: Diarrhea caused significant elevations of trough level, AUC0 4h and Cmax of FK506, and the prolongation of Tmax in renal transplant recipients administered FK506. PMID- 15344966 TI - The microextraction of RNA from archival cardiac allografts embedded in paraffin. AB - One of the difficulties encountered in studying rejection in patients is the availability of tissue. The goal of our study was to isolate RNA from archival allograft tissue, and to demonstrate that it is of suitable quality for further molecular experimentation. Thirty-two paraffin embedded cardiac and five renal allograft archival samples were obtained after IRB approval, from a total of 18 transplant patients (13 cardiac/five renal transplant patients) from a search of the University of Tennessee's teaching hospitals. RNA was extracted from the paraffin blocks and amplified by reverse transcription-polymerase chain reaction (RT-PCR). Grade 3A and higher and non-rejection samples were tested. In addition, normal mouse liver tissue was isolated for comparison. Negative control samples were also included. RT-PCR amplification of 18s RNA, 324 bp target sequences, revealed readily detectable bands. Only one block that was 3 years old did not yield detectable RNA secondary to presumed degradation. The negative control showed no bands at 324 bp. We conclude that RNA from archived allograft tissue can be used for further experiments. The use of this tissue offers some distinct advantages when studying correlation of gene expression with clinical outcome and therapeutic response. PMID- 15344967 TI - Cardiovascular disease in stable renal transplant patients in Norway: morbidity and mortality during a 5-yr follow-up. AB - BACKGROUND: Although cardiovascular disease is a major cause of death after renal transplantation (Tx), predictors for cardiovascular events have not been well defined. Aims of this cross-sectional study were first to assess cardiovascular morbidity and mortality in stable renal Tx patients, and to identify predictors for cardiovascular events during long-term follow-up. METHODS: In all, 406 renal Tx patients (mean age: 47 yr, 60.1% males, 70.9% using cyclosporine A) commenced a baseline registration (median) 48 months after Tx, and 405 was thereafter followed in 5 yr. Kaplan-Meier plots and multivariate regression analysis (Cox proportional hazards model) were used to identify and characterize predictors for cardiovascular events. RESULTS: There were 88 deaths (average annual mortality: 4.4%), and 74% of these were cardiovascular. In age groups 40-49, 50-59, and 60 69 yr, odds ratio for cardiovascular mortality in patients vs. general population was 46.2, 20.1, and 8.0, respectively. Death from ischemic heart disease (IHD) was independently predicted by baseline congestive heart failure (relative risk: RR 5.33), diabetes (RR 2.28), systolic blood pressure (mmHg, RR 1.02), age (yr, RR 1.06), and high-density lipoprotein cholesterol (mmol/L, RR 0.36). Predictors for a major ischemic heart event (death from or onset of IHD) were in addition baseline total cholesterol (mmol/L, RR 1.18) and cerebrovascular disease (RR 2.98). CONCLUSIONS: Thus, IHD was the major cause of death late after renal Tx, and a major ischemic heart event was predicted by baseline congestive heart failure, diabetes, age, hypertension, and hypercholesterolemia. PMID- 15344968 TI - Feasibility of sputum induction in lung transplant recipients. AB - Sputum induction (SI) is nowadays being applied as a non-invasive and safe method to investigate airway inflammation in pulmonary diseases. We investigated the feasibility of SI after lung transplantation (LTX), and compared sputum and bronchoalveolar lavage (BAL) cellular characteristics and interleukin-8 (IL-8) levels. Results were also compared with 11 healthy subjects. SI as performed between 26 and 1947 d after LTX in 19 recipients, was successful in 16 of 22 attempts (73%). Six patients failed to produce sputum after induction, mostly just post-LTX and with having a lower forced expiratory volume in 1 s (FEV1). The success rate in clinically stable patients after the first month post-LTX was 93%. Side-effects were absent. Sputum recovery, viability and squamous cell contamination were comparable between LTX patients and healthy subjects. In the LTX group, total cell counts, neutrophil percentages and IL-8 levels were much higher in SI than BAL (1.6 x 10(6)/mL, 65.5% and 54.2 ng/mL vs. 0.1 x 10(6)/mL, 3.0% and 0.01 ng/mL; p < 0.001). Although LTX-neutrophil percentages in SI and BAL correlated properly (rho=0.72, p=0.04), both techniques are not interchangeable. We conclude that sputum induction is feasible, well tolerated, and without major side-effects in stable patients after the first month post-LTX. Induced sputum may be a useful tool to study inflammatory changes of the airways after LTX, and because of the large quantity of neutrophils sampled, especially for further studies on the pathogenesis of bronchiolitis obliterans. PMID- 15344969 TI - Heel ultrasonography is not a good screening tool for bone loss after kidney and pancreas transplantation. AB - BACKGROUND: Solid organ transplant recipients, particularly simultaneous pancreas kidney recipients, are at high fracture risk. We tested whether quantitative ultrasonography (QUS) of the heel predicts bone mineral density (BMD) by dual energy X-ray absorptiometry (DXA) in solid organ transplant recipients. METHODS: Thirty-eight transplant recipients (22 Female/16 Male) were studied. Spine and hip BMD was measured with a Hologic DXA scanner. 'Stiffness' of the heel was measured with a Lunar Ultrasound densitometer and compared with BMD by DXA. Contributing factors to bone loss were also assessed. RESULTS: Mean age was 43.1 +/- 1.3 yr. Simultaneous pancreas-kidney, kidney, and pancreas alone transplant recipients were assessed. Mean time post-transplantation was 3.0 +/- 0.6 yr. Mean DXA spine T-score was -1.15 +/- 0.22 (mean +/- SEM) and hip T-score was -1.22 +/- 0.20. There was no difference in mean T-score between women and men at the hip or spine. Mean right heel stiffness T-score was -0.97 +/- 0.25. There was no correlation between QUS and DXA at either the hip or spine in women or men. QUS had a false negative rate for identifying osteopenia or osteoporosis of 17% compared with DXA. The false positive rate for identifying osteopenia was 61%. CONCLUSIONS: The QUS is an unacceptable tool for identifying those at risk for bone loss after kidney or pancreas transplantation. PMID- 15344970 TI - Renal transplantation offers a better survival in HCV-infected ESRD patients. AB - Sezer S, Ozdemir FN, Akcay A, Arat Z, Boyacioglu S, Haberal M. Renal transplantation offers a better survival in HCV-infected ESRD patients. Clin Transplant 2004 DOI: 10.1111/j.1399-0012.2004.00252. Abstract: The presence of hepatitis C virus (HCV) infection has been found to adversely affect the morbidity and mortality rates in the dialysis population. Renal transplantation is a treatment option after a careful pre-transplant evaluation. We designed this study to find the impact of HCV infection on patient survival, co-morbidity and allograft survival in a selected group of hemodialysis (HD) and transplant population. We retrospectively analyzed 116 renal transplant patients (94 HCV negative, 22 HCV-positive) and 136 HD patients (106 HCV-negative, 30 HCV positive) who had renal transplantation or underwent dialysis before 1996. The HCV-infected patients were evaluated by liver biopsy for the absence of advanced liver disease before transplantation. There was no clinical or laboratory decompensation of liver disease in transplant and dialysis patient groups. The overall 5-yr survival rates were 85.2% for renal transplant recipients and 74.5% for those on HD. The comparison results revealed a significant difference between HCV-infected patients with and without transplantation. The 3-yr renal allograft survival rates were comparable in HCV-positive and -negative patients, but the risk of chronic allograft nephropathy (CAN) and graft failure were higher at the fifth year in HCV-positive patients. In conclusion, renal transplantation should the preferred therapy in HCV-infected dialysis patients as it improves the survival rates. The presence of HCV infection increases the CAN rate and the influence on allograft survival is evident at the fifth year of assessment. PMID- 15344972 TI - Evidence-based guidelines. PMID- 15344973 TI - Consent for the Aegus: when black is white. PMID- 15344974 TI - Imaging guideline 1: Blunt cervical spine trauma. PMID- 15344975 TI - Fat necrosis in the breast: review of the mammographic and ultrasound features, and a strategy for management. AB - The present study reviews the mammographic and ultrasound features of fat necrosis, the degree of correlation between the two modalities, and proposes a strategy for management, especially in cases with atypical appearances. PMID- 15344976 TI - Role of Tc-glucoheptonic acid brain single photon emission computed tomography in differentiation of recurrent brain tumour and post-radiation gliosis. AB - Blood-brain barrier imaging of brain tumours is fast attracting interest now that it has been demonstrated that disruption of the blood-brain barrier is essential for uptake of all tumour-seeking agents. The aim of the present study was to differentiate recurrent tumour from post-radiation gliosis using (99m)technetium glucoheptonate ((99m)Tc-GHA) as a tumour-seeking agent. Brain single photon emission computed tomography (SPECT) with (99m)Tc-GHA was performed in 73 patients with primary malignant brain tumours after radiotherapy, and the results were correlated with the clinical behaviour of the disease on follow up. The SPECT was suggestive of recurrent tumour in 55 patients. The clinical course was consistent with recurrence in 51 of the 55 patients. The clinical course was consistent with radiation necrosis in the remaining 21 patients, which included 17 patients with a negative SPECT and four patients with a positive SPECT study. Mean GHA index in recurrent tumour and post-radiation gliosis was 7.04 +/- 4.35 and 1.88 +/- 1.70, respectively (P = 0.0001). Mean GHA index in high-grade and low-grade glioma was 7.78 +/- 4.73 and 3.15 +/- 2.44, respectively (P = 0.001). (99m)Technetium-glucoheptonate brain SPECT is a sensitive and reliable diagnostic modality to differentiate recurrent tumour from post-radiation gliosis. PMID- 15344977 TI - Intraductal pancreatic calculi in patients with choledochal cyst. AB - Summary Intraductal pancreatic calculi are a rare association in patients with choledochal cyst (CC). Reported herein is a retrospective analysis of the data on pancreatic calculi in 37 patients (10 children and 27 adults) who underwent endoscopic retrograde cholangiopancreatography (ERCP) for CC over a period of 15 years. Five patients (one child and four adults) were identified as having intraductal pancreatic calculi. Three had type I and two had type IV CC. Four underwent cyst excision. No intervention was carried out for the pancreatic calculi. There was no evidence of pancreatic insufficiency on follow up and none had recurrence of abdominal pain. One patient died of cholangitis, which was the initial presenting problem. Intraductal pancreatic calculi associated with CC, although uncommon, are not rare and are not always symptomatic. PMID- 15344978 TI - Role of mammography in diagnosis of axillary abnormalities in women with normal breast examination. AB - The mammograms of 43 patients presenting with palpable unilateral masses in the axilla and normal breasts on physical examination were retrospectively reviewed to determine the cause and imaging characteristics of axillary abnormalities, and the usefulness of mammography in detecting occult breast carcinoma. Cytological or histological confirmation was obtained in all patients. Forty of 43 patients had axillary lymphadenopathy while three had lipoma, fibroadenoma and haematoma, respectively. Causes of malignant lymphadenopathy (n = 22) were metastatic diseases from non-mammary primary malignancy (n = 8), occult ipsilateral breast carcinoma (n = 5), and previous contralateral breast carcinoma (n = 9). Causes of benign lymphadenopathy (n = 18) were reactive nodal hyperplasia (n = 6), collagen vascular diseases (n = 2), and acute bacterial (n = 2) and tuberculous (n = 8) lymphadenitis. Nodal size was not significantly different between benign and malignant lymph nodes. Benign and malignant nodal margins were variable. Intranodal microcalcifications were found in two cases of breast carcinoma metastasis. Intranodal macrocalcifications were found in three cases of tuberculous lymphadenitis. Occult primary breast carcinoma was detected on mammograms in four of five patients with axillary lymphadenopathy due to ipsilateral breast carcinoma. Mammographical features of benign and malignant lymphadenopathy may be indistinguishable, but presence of intranodal calcifications is helpful. Mammography is also valuable in depicting occult primary breast carcinoma. PMID- 15344979 TI - Embolization for major lower gastrointestinal haemorrhage: five-year experience. AB - The management of major lower gastrointestinal haemorrhage has changed dramatically in the last 15 years. Innovations in coaxial catheter technology have allowed the interventional radiologist to reach the small peripheral mesenteric arteries and perform superselective embolization with a variety of agents. The present large series represents the 5-year experience of this technique at the Alfred Hospital, Melbourne, in a patient cohort with a high number of comorbidities. Technical success was achieved in 96% of cases. The clinical symptoms of mesenteric ischaemia developed in four patients after embolization and were managed conservatively in two. The procedure-related mortality was low when compared with the published complication rates for emergency surgery, in this clinical setting. PMID- 15344980 TI - Computed tomography evaluation of cavitary necrosis in complicated childhood pneumonia. AB - The purpose of the present study was to retrospectively investigate the chest radiograph (CR) and CT findings of childhood pneumonia complicated by cavitary necrosis, and to evaluate the role of CT in decision-making for surgical intervention. Chest CT was performed in 51 patients presenting with persistent or progressive pneumonia, respiratory distress and sepsis despite 7-10 days of appropriate antibiotic treatment and closed tube drainage. Chest radiograph and CT findings were retrospectively evaluated in 23 patients (45%) with cavitary necrosis. Chest radiographs showed consolidation in 19 of 23 patients, cavitation in five patients, parapneumonic effusions in 17 patients and air-fluid levels in the pleural space in one patient. The CT scans demonstrated consolidation and cavitary necrosis in all patients. There were parapneumonic effusions in all patients with concomitant loculated collections in six patients. Twenty-two of 23 patients had pleural thickening. In seven patients there were air-fluid levels in the pleural space. In five of these patients, CT scans demonstrated bronchopleural fistulae. On the basis of the CT and clinical findings, 11 patients underwent surgical intervention. Computed tomography is superior to CR for demonstrating cavitary necrosis complicating pneumonia, and other parenchymal and pleural complications. It also has a crucial decision-making role for surgery. PMID- 15344981 TI - Perfusion magnetic resonance imaging and magnetic resonance spectroscopy of cerebral gliomas showing imperceptible contrast enhancement on conventional magnetic resonance imaging. AB - The purpose of the present paper was to evaluate the utility of perfusion MRI in cerebral gliomas showing imperceptible contrast enhancement on conventional MRI, and to evaluate the relationships of perfusion MRI and magnetic resonance (MR) spectroscopic results in these tumours. Twenty-two patients with histopathologically proven cerebral gliomas and showing insignificant contrast enhancement on conventional MR were included in the present study. All patients underwent perfusion MRI and MR spectroscopy on a 1.5-T MR system. Significant differences of the relative cerebral blood volume (rCBV) values and the choline : creatine ratios were noted between low-grade and anaplastic gliomas (P < 0.01). Good correlation was found between the rCBV values and the choline : creatine values (y = 0. 532x + 1.5643; r = 0.67). Perfusion MRI can be a useful tool in assessing the histopathological grade of non-contrast-enhancing cerebral gliomas. Along with MR spectroscopic imaging it can serve as an important technique for preoperative characterization of such gliomas, so that accurate targeting by stereotactic biopsies is possible. PMID- 15344982 TI - New endoluminal technology: efficacy and safety--whose responsibility? AB - New endoluminal technology is being increasingly used in the management of arterial disease. With this increased usage of often experimental devices there is a growing responsibility for clinicians to collect data and report adverse events particularly in the longer term, when initial feasibility studies have been concluded. Many of these devices will have a perceived advantage so great that randomized controlled trials are not feasible. It is important that these perceptions are based on robust data not clinical impressions. The following article discusses methods of device assessment and looks at who is responsible for ongoing device evaluation. PMID- 15344983 TI - Problem-based learning and medical student radiology teaching. AB - The present study describes some practical advantages and disadvantages of problem-based learning (PBL), and makes suggestions to improve PBL radiology teaching of medical students. PMID- 15344984 TI - Radiation-induced brain disorders in patients with pituitary tumours. AB - Radiation-induced brain disorders (RIBD) are uncommon and they are grave sequelae of conventional radiotherapy. In the present report, we describe the clinical spectrum of RIBD in 11 patients who received post-surgery conventional megavoltage irradiation for residual pituitary tumours. Of these 11 patients (nine men, two women), seven had been treated for non-functioning pituitary tumours and four for somatotropinomas. At the time of irradiation the age of these patients ranged from 30 to 59 years (mean, 39.4 +/- 8.3; median, 36) with a follow-up period of 6-96 months (mean, 18.3 +/- 26.4; median, 11). The dose of radiation ranged from 45 to 90 Gy (mean, 51.3 +/- 13.4; median, 45), which was given in 15-30 fractions (mean, 18.6 +/- 5.0; median, 15) with 2.8 +/- 0.3 Gy (median, 3) per fraction. The biological effective dose calculated for late complications in these patients ranged from 78.7 to 180 Gy (mean, 99.1 +/- 27.5; median, 90). The lag time between tumour irradiation and the onset of symptoms ranged from 6 to 168 months (mean, 46.3 +/- 57.0; median, 57). The clinical spectrum of RIBD included new-onset visual abnormalities in five, cerebral radionecrosis in the form of altered sensorium in four, generalized seizures in four, cognitive dysfunction in five, dementia in three and motor deficits in two patients. Magnetic resonance imaging (MRI)/CT of the brain was suggestive of radionecrosis in eight, cerebral oedema in three, cerebral atrophy in two and second neoplasia in one patient. Associated hormone deficiencies at presentation were hypogonadism in eight, hypoadrenalism in six, hypothyroidism in four and diabetes insipidus in one patient. Autopsy in two patients showed primitive neuroectodermal tumour (PNET) and brainstem radionecrosis in one, and a cystic lesion in the left frontal lobe following radionecrosis in the other. We conclude that RIBD have distinctive but varying clinical and radiological presentations. Diabetes insipidus and PNET as a second neoplastic disorder in adults following pituitary irradiation have not been reported previously. PMID- 15344985 TI - Multicentre analysis of treatment planning information: technical requirements, possible applications and a proposal. AB - Digital data from 3-D treatment planning computers is generally used for patient planning and then never considered again. However, such data contains enormous quantities of information regarding patient geometries, tissue outlining, treatment approaches and dose distributions. Were such data accessible from planning systems from multiple manufacturers, there would be substantial opportunities for undertaking quality assurance of radiotherapy clinical trials, prospective assessment of trial outcomes and basic treatment planning research and development. The technicalities of data exchange between planning systems are outlined, and previous attempts at producing systems capable of viewing and/or manipulating imaging and radiotherapy digital data reviewed. Development of a software system for enhancing the quality of Australasian clinical trials is proposed. PMID- 15344986 TI - Do written consent forms provide medicolegal protection from litigation in radiotherapy? AB - Two methods were used in an attempt to determine whether written consent forms serve a useful medicolegal purpose in Australian radiotherapy departments. First a survey was posted to all practising radiation oncologists to ask about their use of written consent forms and whether they had ever been useful in any claims made against them. Second, the largest medical defence organization in Australia was contacted for de-identified details about every claim ever made against a radiation oncologist. The survey yielded a response rate of 50.3%. Only one respondent indicated that any consent form had ever played a role in any claim and was not prepared to discuss it. Many radiation oncologists did not routinely use a consent form and its use could not be considered standard practice. The submitted consent forms generally did not contain warnings to the patient about specific risks of treatment. The details of the previous claims provided by the medical defence organization indicated that more often claims arose from circumstances in which a written consent form could not provide any useful protection such as radiation dose calculation errors. The medicolegal value of written consent forms in radiotherapy seems low, although further research may be necessary before dispensing with them completely. PMID- 15344987 TI - Acute radiation toxicity assessment of a 3-D conformal head and neck radiation treatment technique. AB - Patients undergoing radiation treatment for head and neck cancer have many critical structures within or adjacent to the treatment area. Avoiding these critical structures is more efficient and easier using 3-D conformal planning. At Sir Charles Gairdner Hospital an investigation into the acute radiation toxicity was undertaken when the head and neck treatment technique was changed from the conventional 2-D plan to the conformal 3-D plan. Although the primary target volume (PTV), fraction size and overall dose remain the same for the two techniques, differing beam configurations mean that treatment fields enter and exit through different parts of the head and neck. Ten patients were initially assessed to gain a baseline appearance of their head and neck region, and then graded weekly according to the toxicity criteria during treatment and at 4, 8 and 12 weeks after treatment. The results of the assessment indicate that there is no increase in toxicity as a result of treating using the conformal head and neck technique. PMID- 15344988 TI - Assessing outcomes of cancer care: lessons to be learned from a retrospective review of the management of small cell lung cancer at the Cancer Therapy Centre, Liverpool Hospital, January 1996-July 2000. AB - The purpose of the present paper was to review the outcomes of care of small cell lung cancer (SCLC) at one Sydney teaching hospital. A retrospective cohort study was carried out of patients with SCLC seen between January 1996 and July 2000. The main outcomes were relapse-free and overall survival. Secondary outcomes of interest were the uniformity of staging investigations, initial treatment, use of prophylactic cranial irradiation (PCI), patterns of relapse and treatment received following relapse. One hundred and three patients with SCLC were treated at the Liverpool Hospital Cancer Therapy Centre during this period. There were 58 men (56%) and 45 women (44%). Forty-two patients (41%) had limited stage disease (LD) and 61 (59%) had extensive stage disease (ED). There was considerable variation in staging investigations. There was little variation in systemic treatment of SCLC. Only 32 of 42 patients with limited stage SCLC were candidates for thoracic radiotherapy and only seven patients received PCI. Median relapse free survival was 11.2 months (95% confidence interval (CI): 7.7-14.8) for patients with LD and 6 months (95%CI: 4.4-7.5) for ED. Median overall survival was 15.1 months (95%CI: 11-19.1) for patients with LD and 8.9 months (95%CI: 7.5 10.2) for ED. Some health outcomes similar to that reported in clinical trials can be achieved in clinical practice. Measuring health outcomes is an important process of maintaining quality of care. PMID- 15344989 TI - All delays before radiotherapy risk progression of Merkel cell carcinoma. AB - Prolonged waiting times for radiotherapy have resulted in many centres assigning priorities to various patient or diagnostic groups. A high risk of progression on a waiting list is one factor that would reasonably influence the priority. The present descriptive study of 27 patients with Merkel cell carcinoma (MCC) found that a median wait of 24 days for radiotherapy is associated with a high risk of progression. Eleven (41%) of 27 patients developed progressive disease, including five (45%) of 11 patients waiting for adjuvant radiotherapy. Patients treated adjuvantly also had longer waiting times prior to their initial radiotherapy consultation (median 41 days), which may have contributed to the rate of progression. Merkel cell carcinoma is an aggressive but curable malignancy and appropriate management should include efforts to minimize all potential delays prior to the commencement of radiotherapy. PMID- 15344990 TI - Breast-conserving therapy in young women with invasive carcinoma of the breast. AB - Higher local recurrence rates have been reported in young women with invasive carcinoma of the breast treated with breast-conserving therapy (BCT). However, age itself may not be responsible for this increased risk of recurrence. To investigate this further, a computerized literature search of MEDLINE was performed using data from 1996 to May 2003. The research was limited to female patients with localized, invasive adenocarcinoma of the breast but also included patients of young age with ductal carcinoma in situ. Women of young age with breast cancer, treated with BCT are at an increased risk of recurrence ranging from 7.5 to 35%. However, the data would suggest that the increased risk is secondary to the association of young age with more aggressive tumours and a positive family history of breast cancer. Other factors that may explain the adverse prognosis in women of a young age include associated genetic abnormalities and the lack of mammographic screening programmes for women of young age. Young age is a risk factor for breast recurrence after BCT. However, management decisions should be based on tumour stage, grade and other related prognostic features rather than on young age alone. PMID- 15344991 TI - Cardiac metastasis from carcinoid tumour: magnetic resonance imaging findings. AB - A case of myocardial metastasis from a gastrointestinal carcinoid is presented. The colon was the primary site. The patient did not manifest symptoms of carcinoid syndrome and had no echocardiographic or MRI evidence of carcinoid valvular heart disease. PMID- 15344992 TI - Atypical diffusion-weighted magnetic resonance findings in glioblastoma multiforme. AB - The present report describes two patients with glioblastoma multiforme with the tumour demonstrating low central apparent diffusion coefficient values similar to those found in cerebral abscesses. Although conventional MR images were fairly specific for tumour, the cases illustrate the need for exercising caution when using diffusion-weighted (DW) MR images for the differentiation of necrotic brain tumours from abscesses. The DW MR imaging information should always be integrated with the conventional spin-echo MR images. PMID- 15344993 TI - Accurate localization of supernumerary mediastinal parathyroid adenomas by a combination of structural and functional imaging. AB - Reoperation for refractory or recurrent hyperparathyroidism following parathyroidectomy carries the potential for increased morbidity and the possibility of failure to localize and remove the lesion intraoperatively. Reported herein are three cases demonstrating the combined use of sestamibi scintigraphy, CT and MR for accurate localization of mediastinal parathyroid adenomas. PMID- 15344994 TI - Bladder pheochromocytoma encountered on sonography. AB - Pheochromocytomas of the bladder are rare neoplasms, constituting < 0.06% of all vesical tumours. Common presenting features of this tumour include episodes of sweating, hypertension, haematuria and postmicturition syncope. We describe a case of bladder pheochromocytoma in a 66-year-old man whose only symptom of macroscopic haematuria was initially assessed with ultrasonography. Clinical presentation highlights the need for a high index of suspicion during sonographic evaluation of bladder neoplasms because such tumours might present without symptoms of adrenergic excess. PMID- 15344995 TI - Role of embolization in the management of splenic trauma. AB - The management of splenic trauma is determined by factors related to a patient's clinical state. Currently, non-surgical management is favoured because of morbidity related to laparotomy and splenectomy. However, the presence of a pseudoaneurysm might necessitate selective splenic arteriography and embolization. Even though this also has associated morbidity, conservation of viable splenic tissue is a desired result. PMID- 15344996 TI - Sarcoidosis of the thoracic and lumbar spine: imaging findings with an emphasis on magnetic resonance imaging. AB - Vertebral sarcoidosis is exceedingly rare with only a few cases reported in the published literature. The case of a 40-year-old man with vertebral sarcoidosis is presented, emphasizing the role of MRI and biopsy in confirming the diagnosis. A brief review of the published literature is also presented. PMID- 15344997 TI - Cherubism: review of four affected members in a Kuwaiti family. AB - Cherubism is a rare autosomal dominant fibro-osseous disorder of childhood, mostly limited to the maxilla and mandible. Extra-cranial skeletal involvement is rare. Post-pubertal involution of the process and jaw remodeling occurs in adulthood. The aim of this case report is to present four cherubs, a father and his three daughters, in whom the diagnosis was made based on history, physical examination, laboratory tests and typical radiological features. The imaging characteristics are discussed and the published literature is reviewed. PMID- 15344998 TI - Concurrent emphysematous pyelonephritis and emphysematous cholecystitis in type 2 diabetes. AB - Concurrence of emphysematous pyelonephritis and emphysematous cholecystitis in a patient with type 2 diabetes has not been reported in the published literature, to the authors' knowledge. A patient who had both at the time of presentation is reported. The mechanism of gas formation and the management strategies are discussed. PMID- 15344999 TI - Ultrasound and magnetic resonance imaging features in a patient with eosinophilic fasciitis. AB - Eosinophilic fasciitis is a rare rheumatological disorder. We present a case of a young male patient who presented with this disease, and we describe how the findings on ultrasound and MRI correlated with the underlying pathology and assisted the diagnosis and the management of the patient. PMID- 15345000 TI - Direct thrombin injection into aneurysmal sac in a patient with a type II endoleak. AB - Endovascular grafting of abdominal aortic aneurysms provides a good alternative to open surgery, especially in high-risk patients. Endoleaks are a well recognized complication and are typically diagnosed on CT. We describe a case in which a patient's endoleak was evaluated by MRI and successfully treated by direct thrombin injection into the site of the leak. PMID- 15345001 TI - Primary papillary serous carcinoma of the peritoneum: four cases and review of computed tomography findings. AB - Primary papillary serous carcinoma of the peritoneum is an uncommon primary malignancy of the peritoneum and is histologically indistinguishable from papillary serous carcinoma of the ovary. The diagnosis of primary peritoneal papillary serous carcinoma should be considered in the presence of peritoneal and omental masses in the absence of an ovarian mass. Although it has been extensively documented in the pathological and gynaecological oncology literature, the CT appearance of primary papillary serous carcinoma of the peritoneum has been reported in only 51 cases in five reports. We present four patients with CT findings of pathologically proven primary papillary serous carcinoma of the peritoneum. There were a total of 23 patients with a histopathologically proven diagnosis of primary papillary serous carcinoma of the peritoneum between 1980 and 2002 with CT imaging. However, only four of the 23 patients' CT films were retrieved for retrospective evaluation. The rest of the films were not available as either patients had misplaced the films or patients were deceased. PMID- 15345002 TI - Angioplasty and stenting of the superior mesenteric artery in acute mesenteric ischaemia. AB - Acute mesenteric ischaemia is a challenging diagnostic problem with a very high mortality. Traditionally, laparotomy is required for definitive management. We describe a successful case of angioplasty and stenting of the superior mesenteric artery in a surgically unfit patient. We recommend that stenting be considered only in situations where the diagnosis has been made prior to bowel infarction, and if the patient poses a poor operative risk. PMID- 15345003 TI - Cerebellar metastases from prostatic carcinoma. AB - A 67-year-old man presented with worsening headaches and gait disturbance, and with minimal clinical signs. A brain MRI showed multiple solid cerebellar metastases. The pathological diagnosis of metastatic carcinoma of the prostate was further suggested by an elevation in prostate specific antigen, and was pathologically confirmed following the neurosurgical removal of the tumours. The rarity of this presentation, as documented in the clinical literature, is reviewed. PMID- 15345004 TI - Charles Stuart (1917-2003). PMID- 15345005 TI - Ross Richard Bowey (1925-2004). PMID- 15345007 TI - RE: Viewing of lateral radiographs. PMID- 15345006 TI - RE: C Acton et al.--Audit of neonatal renal scans. PMID- 15345008 TI - RE: Cranial CT interpretation by senior emergency department staff. PMID- 15345012 TI - Philanthropy and prosthodontics. PMID- 15345009 TI - RE: How safe is Australian radiotherapy? PMID- 15345013 TI - Biaxial flexural strength and microstructure changes of two recycled pressable glass ceramics. AB - PURPOSE: This study evaluated the biaxial flexural strength and identified the crystalline phases and the microstructural features of pressed and repressed materials of the glass ceramics, Empress 1 and Empress 2. MATERIALS AND METHODS: Twenty pressed and 20 repressed disc specimens measuring 14 mm x 1 mm per material were prepared following the manufacturers' recommendations. Biaxial flexure (piston on 3-ball method) was used to assess strength. X-ray diffraction was performed to identify the crystalline phases, and a scanning electron microscope was used to disclose microstructural features. RESULTS: Biaxial flexural strength, for the pressed and repressed specimens, respectively, were E1 [148 (SD 18) and 149 (SD 35)] and E2 [340 (SD 40), 325 (SD 60)] MPa. There was no significant difference in strength between the pressed and the repressed groups of either material, Empress 1 and Empress 2 (p > 0.05). Weibull modulus values results were E1: (8, 4.7) and E2: (9, 5.8) for the same groups, respectively. X ray diffraction revealed that leucite was the main crystalline phase for Empress 1 groups, and lithium disilicate for Empress 2 groups. No further peaks were observed in the X-ray diffraction patterns of either material after repressing. Dispersed leucite crystals and cracks within the leucite crystals and glass matrix were features observed in Empress 1 for pressed and repressed samples. Similar microstructure features--dense lithium disilicate crystals within a glass matrix--were observed in Empress 2 pressed and repressed materials. However, the repressed material showed larger lithium disilicate crystals than the singly pressed material. CONCLUSIONS: Second pressing had no significant effect on the biaxial flexural strength of Empress 1 or Empress 2; however, higher strength variations among the repressed samples of the materials may indicate less reliability of these materials after second pressing. PMID- 15345014 TI - An experimental study on particular physical properties of several interocclusal recording media. Part II: Linear dimensional change and accompanying weight change. AB - PURPOSE: The purpose of this study was to evaluate the linear dimensional change and accompanying weight change of several elastomeric interocclusal recording media. MATERIALS AND METHODS: Ten 50 mm long, 8 mm wide, 3 mm deep Teflon molds and a stainless steel die were constructed for the purposes of this study. The die was made in order to produce standardized reference lines, which were used for the measurement of the linear change. After homogeneous mixing, each material was carried from the mixing pad or directly from the mixing tip to the Teflon mold. The mold was inverted onto the stainless steel die. The materials were allowed to set for the manufacturers' suggested setting time plus an additional 3 minutes to ensure polymerization of the material. An electronic scale (Galaxy 110, Ohaus, Pine Brook, NJ) was used for the measurement of the weight change of the specimens and a traveling micrometer microscope (Griffin Ltd., London, England) was used to measure the linear changes at 5 time intervals: 0, 1 hour, 24, 48, and 72 hours. RESULTS: Analysis of variance for a significance level of 5% revealed that there was a statistically significant effect of the "material" factor on the weight changes (F= 2,229.98, p < 0.0005). There was also a statistically significant effect of the "time" factor on the weight changes (F= 2,332.04, p < 0.0005). Descriptive statistics and Tukey's Honest Significant Difference Test revealed that the elastomeric material with the greatest weight changes is polyether (Ramitec). Analysis of variance for a significance level of 5% revealed that there is a statistically significant effect of the "material" factor on the linear changes (F= 215.54, p < 0.0005). There is also a statistically significant effect of the "time" factor on the linear changes (F= 1,996.01, p < 0.0005). Descriptive statistics and Tukey's Honest Significant Difference test revealed that the elastomeric material with the smallest linear changes is polyether (Ramitec). CONCLUSIONS: Of all materials tested, Ramitec (polyether) presented the smallest linear change at all time intervals. Addition reaction silicones presented statistically significant differences in recordings of linear changes among them only at the 1st and the 24th hour. Linear changes did not seem to be associated with weight changes. PMID- 15345015 TI - Patient evaluation of treatment with fixed prostheses supported by implants or a combination of teeth and implants. AB - PURPOSE: The objective of this study was to compare treatment outcomes among subjects with complete arch fixed prostheses in the maxilla, supported by implants or a combination of natural teeth and dental implants. MATERIALS AND METHODS: Twenty-one subjects with maxillary tooth- and implant-supported fixed prostheses and 21 subjects with maxillary implant-supported fixed prostheses were identified and included in the study. All abutment teeth in the group with tooth- and implant-supported prostheses were provided with cemented copings that incorporated threads for vertical locking screws. Frameworks were fabricated with a gold alloy that was veneered with acrylic resin or ceramic materials. All frameworks were screw-retained to implants and copings. Frameworks in the group with implant-supported prostheses were fabricated with milled titanium or gold alloy to which denture teeth and resin base material were applied. All prostheses had a minimum of 8 units, at least 4 of which were in one quadrant. Subjects in both groups were mailed a questionnaire consisting of 15 questions focused on various factors related to treatment outcome, such as oral function and patient satisfaction. RESULTS: The response rate was 86%. Both groups reported a high satisfaction rate for most items with few regretting their choice of treatment. Most individuals in both groups reported great improvement in chewing ability and few reported phonetic disturbances. No statistically significant differences were found between the groups. CONCLUSION: The results of the present study showed similarity in questionnaire responses between the 2 groups of participants. High satisfaction was reported both among subjects who received a complete arch fixed prosthesis in the maxilla supported by dental implants only, as well as among those whose prostheses were supported by a combination of natural teeth and dental implants. PMID- 15345017 TI - Effect of denture cleansers on the surface roughness and hardness of a microwave cured acrylic resin and dental alloys. AB - PURPOSE: This study evaluated the effect of denture cleansers on the surface hardness of a denture base resin, and on the surface roughness of the resin and Co-Cr and Ti-6Al-4V alloys. MATERIALS AND METHODS: Forty-eight disc-shaped specimens were fabricated of a microwave-cured acrylic resin, each having one of the alloys attached to its surface. The specimens were randomly divided into 6 groups, each consisting of 8 samples. Specimens were exposed to one of the three cleansing treatments (polident, manipulation pharmacy cleanser, and water) as follows. Group I: Co-Cr + polident; Group II: Co-Cr + manipulation; Group III: Co Cr + water; Group IV: Ti-6Al-4V + polident; Group V: Ti-6Al-4V + manipulation; and Group VI: Ti-6Al-4V + water. Three exposures lasting 5 minutes each were conducted daily, and repeated after storage periods of 1, 14, and 29 days in artificial saliva at 37 degrees C. Hardness and roughness measurements were undertaken immediately after specimen preparation (T0) and on the 1st (T1), 15th (T15), and 30th (T30) day following the beginning of storage. Three roughness and hardness evaluations were carried out for each sample and testing time, and mean values were calculated. Results were analyzed using ANOVA and linear regression. RESULTS: The Knoop hardness test demonstrated differences (p < 0.05) between Groups I and IV at T1 and T30 (14.30 +/- 2.78; 14.06 +/- 1.76) and between Groups II and V at T15 (16.99 +/- 2.24). Significant differences (p < 0.05) in resin roughness (in microm) were observed between Groups I and IV at T15 and T30 (0.14 +/- 0.06; 0.21 +/- 0.38). With regard to Co-Cr, roughness data showed differences (p < 0.05) for all groups at T30 (Group I: 0.15 +/- 0.07; Group II: 2.43 +/- 0.66; Group III: 4.05 +/- 1.03), for Group II at T1 (0.10 +/- 0.03), and for Group I at T15 (0.15 +/- 0.02). There were significant differences (p < 0.05) in titanium roughness for Group IV at T15 (0.12 +/- 0.01) and T30 (0.11 +/- 0.04). CONCLUSIONS: Manipulated cleanser containing sodium perborate increased surface roughness and hardness, probably due to its incapacity to remove the pellicle formed on the acrylic resin and dental alloys. PMID- 15345016 TI - The effects of a commercial aluminum airpolishing powder on dental restorative materials. AB - INTRODUCTION: JET-Fresh (Dentsply International, York, PA), a new airpolishing powder that contains aluminum trihydroxide as the abrasive agent, has been introduced for use with Prophy-Jet trade mark (Dentsply International). PURPOSE: The aim of this study was to investigate the surface effects of aluminum trihydroxide airpolishing powder on a series of restorative materials. MATERIALS AND METHODS: A total of 6 Class V preparations with enamel margins were created on the buccal surface of extracted human molars. The preparations were restored with Type III gold, a high copper spherical alloy amalgam, porcelain, a light activated hybrid composite material, a light-activated microfilled composite material, and a light-activated, resin-modified glass ionomer material. The distal half of each restoration was covered with fiberglass tape and the mesial half subjected to treatment with the aluminum trihydroxide powder via the Prophy Jet for 5 seconds. After removal of the protective tape, the buccal surface of each treated tooth was replicated with impression material. Replicas were generated using epoxy resin and prepared for evaluation with scanning electron microscopy. RESULTS: The aluminum trihydroxide produced surface alterations that were apparent visually and when viewed by a scanning microscope of the hybrid and microfilled composites and the glass ionomer restorations. The surfaces of the amalgam and gold restorations were altered, but not to the extent that the resin based materials were. No disruption of the surface characterization of the porcelain was detected; however, with gold and porcelain materials, the aluminum trihydroxide removed notable amounts of the luting cements (the results are consistent with the data gathered with sodium bicarbonate powder). CONCLUSION: Aluminum trihydroxide as the abrasive agent in an airpolishing system should be avoided on resin composites, resin-modified composites, and around the margins of cemented restorations. PMID- 15345018 TI - Fabrication of a maxillofacial prosthesis using a computer-aided design and manufacturing system. AB - PURPOSE: Maxillofacial prostheses are usually fabricated on the basis of impressions made with dental-impression material. The extent to which the prosthesis reproduces normal facial morphology depends on the clinical judgment of the individual fabricating the prosthesis. This paper describes a computer aided design and manufacturing (CAD/CAM) system for the fabrication of maxillofacial prostheses. This system will provide a more consistently accurate reproduction of facial morphology. MATERIALS AND METHODS: Facial measurements were taken using a non-contact three-dimensional laser morphological measurement system. The measurements were sent to a computer numerical controlled (CNC) milling machine to generate a cast of the patient's face for the fabrication of prosthesis. RESULTS: Facial contours were measured using a laser. This method minimizes patient discomfort and avoids soft tissue distortion by impression material. Moreover, the digital data obtained is easy to store and transmit, and mirror-images can be readily generated by computer processing. CONCLUSION: This method offers an objective, quantified approach for fabricating maxillofacial prostheses. PMID- 15345019 TI - Mechanical failure of an implant-retained bar: a clinical report. AB - This clinical report describes the mechanical failure of an implant-retained bar where the bar fractured midway between two implants. The location of the fracture is not typical for implant-retained bars. Possible causes of the fracture are discussed. The prosthodontic treatment provided to manage the mechanical failure and prevent it from recurring is also discussed. PMID- 15345020 TI - Implant-supported rehabilitation of the severely atrophic maxilla: a clinical report. AB - Implant-supported rehabilitation in the posterior maxillary region presents a series of challenges because of the possibility of increased pneumatization of the maxillary sinus after dental loss. In cases where significant maxillary bone resorption has occurred, efforts center on maximizing the use of the remaining bone to afford primary stability to the implants. This clinical report describes a patient with extreme maxillary atrophy for whom fixed rehabilitation supported on 8 implants was accomplished. A zygomatic implant, a pterygomaxillary implant, and 2 implants mesial and distal to both canine eminences were placed. This rehabilitation was accomplished with a less invasive technique and in a much shorter time period compared to a sinus lift procedure. One year after prosthetic loading, the clinical and radiological results remain satisfactory. PMID- 15345021 TI - Simple(r) soft tissue masque for individual fixed restorations. AB - This article describes a simplified technique for creating a soft tissue masque to aid in shaping the gingival contours of fixed restorations. The additional clinical visit that has been previously described is circumvented, and only one master cast is required. PMID- 15345022 TI - The history and development of the dental surveyor--Part III. PMID- 15345027 TI - Survival of patients with metastatic breast cancer: twenty-year data from two SEER registries. AB - BACKGROUND: Many researchers are interested to know if there are any improvements in recent treatment results for metastatic breast cancer in the community, especially for 10- or 15-year survival. METHODS: Between 1981 and 1985, 782 and 580 female patients with metastatic breast cancer were extracted respectively from the Connecticut and San Francisco-Oakland registries of the Surveillance, Epidemiology, and End Results (SEER) database. The lognormal statistical method to estimate survival was retrospectively validated since the 15-year cause specific survival rates could be calculated using the standard life-table actuarial method. Estimated rates were compared to the actuarial data available in 2000. Between 1991 and 1995, further 752 and 632 female patients with metastatic breast cancer were extracted respectively from the Connecticut and San Francisco-Oakland registries. The data were analyzed to estimate the 15-year cause-specific survival rates before the year 2005. RESULTS: The 5-year period (1981-1985) was chosen, and patients were followed as a cohort for an additional 3 years. The estimated 15-year cause-specific survival rates were 7.1% (95% confidence interval, CI, 1.8-12.4) and 9.1% (95% CI, 3.8-14.4) by the lognormal model for the two registries of Connecticut and San Francisco-Oakland respectively. Since the SEER database provides follow-up information to the end of the year 2000, actuarial calculation can be performed to confirm (validate) the estimation. The Kaplan-Meier calculation for the 15-year cause-specific survival rates were 8.3% (95% CI, 5.8-10.8) and 7.0% (95% CI, 4.3-9.7) respectively. Using the 1991-1995 5-year period cohort and followed for an additional 3 years, the 15-year cause-specific survival rates were estimated to be 9.1% (95% CI, 3.8-14.4) and 14.7% (95% CI, 9.8-19.6) for the two registries of Connecticut and San Francisco-Oakland respectively. CONCLUSIONS: For the period 1981-1985, the 15-year cause-specific survival for the Connecticut and the San Francisco-Oakland registries were comparable. For the period 1991-1995, there was not much change in survival for the Connecticut registry patients, but there was an improvement in survival for the San Francisco-Oakland registry patients. PMID- 15345028 TI - TM4SF10 gene sequencing in XLMR patients identifies common polymorphisms but no disease-associated mutation. AB - BACKGROUND: The TM4SF10 gene encodes a putative four-transmembrane domains protein of unknown function termed Brain Cell Membrane Protein 1 (BCMP1), and is abundantly expressed in the brain. This gene is located on the short arm of human chromosome X at p21.1. The hypothesis that mutations in the TM4SF10 gene are associated with impaired brain function was investigated by sequencing the gene in individuals with hereditary X-linked mental retardation (XLMR). METHODS: The coding region (543 bp) of TM4SF10, including intronic junctions, and the long 3' untranslated region (3 233 bp), that has been conserved during evolution, were sequenced in 16 male XLMR patients from 14 unrelated families with definite, or suggestive, linkage to the TM4SF10 gene locus, and in 5 normal males. RESULTS: Five sequence changes were identified but none was found to be associated with the disease. Two of these changes correspond to previously known SNPs, while three other were novel SNPs in the TM4SF10 gene. CONCLUSION: We have investigated the majority of the known MRX families linked to the TM4SF10 gene region. In the absence of mutations detected, our study indicates that alterations of TM4SF10 are not a frequent cause of XLMR. PMID- 15345029 TI - A robust, low- to medium-throughput prnp genotyping system in sheep. AB - BACKGROUND: In many countries breeding programs for resistance to scrapie in sheep are established. Therefore, the demand on genotyping capacities of the polymorphisms of the prion protein gene (prnp) relevant to presently known disease associations and EU regulations is steadily increasing. Most published typing methods are not well suited for routine typing of large sample numbers in smaller service laboratories for different reasons: they require partly manual data processing, sophisticated and sensitive protocols, high efforts regarding time and manpower, multiple step reactions or substantial hardware investments. To overcome these drawbacks, we developed a prnp typing method that is based on a 'multiplex amplification refractory mutation system' (ARMS) reaction. METHODS: In this study we combined the amplification refractory mutation system (ARMS) with standard fluorescent based fragment length analyses method to develop a prnp genotyping method (PRNP ARMS). RESULTS: By optimised primer design it was possible to type the 4 relevant single nucleotide polymorphisms (SNPs) in the prnp simultaneously in one multiplex reaction. Automated fragment length analysis enabled automated allele designation. Suitability of the PRNP ARMS for routine application was proven by typing samples with known genotypes and larger sample numbers from half-sib families. CONCLUSION: The ARMS PRNP typing method established in this study is universally suited for a broad range of typing projects with different requirements. It provides an efficient and inexpensive diagnostic mutation analysis that will improve the quality of prnp genotyping compared with other low-cost methods. It can be implemented by most molecular genetic laboratories using standard equipment. PMID- 15345030 TI - Exonuclease activity and P nucleotide addition in the generation of the expressed immunoglobulin repertoire. AB - BACKGROUND: Immunoglobulin rearrangement involves random and imprecise processes that act to both create and constrain diversity. Two such processes are the loss of nucleotides through the action of unknown exonuclease(s) and the addition of P nucleotides. The study of such processes has been compromised by difficulties in reliably aligning immunoglobulin genes and in the partitioning of nucleotides between segment ends, and between N and P nucleotides. RESULTS: A dataset of 294 human IgM sequences was created and partitioned with the aid of a probabilistic model. Non-random removal of nucleotides is seen between the three IGH gene types with the IGHV gene averaging removals of 1.2 nucleotides compared to 4.7 for the other gene ends (p < 0.001). Individual IGHV, IGHD and IGHJ gene subgroups also display statistical differences in the level of nucleotide loss. For example, within the IGHJ group, IGHJ3 has average removals of 1.3 nucleotides compared to 6.4 nucleotides for IGHJ6 genes (p < 0.002). Analysis of putative P nucleotides within the IgM and pooled datasets revealed only a single putative P nucleotide motif (GTT at the 3' D-REGION end) to occur at a frequency significantly higher then would be expected from random N nucleotide addition. CONCLUSIONS: The loss of nucleotides due to the action of exonucleases is not random, but is influenced by the nucleotide composition of the genes. P nucleotides do not make a significant contribution to diversity of immunoglobulin sequences. Although palindromic sequences are present in 10% of immunologlobulin rearrangements, most of the 'palindromic' nucleotides are likely to have been inserted into the junction during the process of N nucleotide addition. P nucleotides can only be stated with confidence to contribute to diversity of less than 1% of sequences. Any attempt to identify P nucleotides in immunoglobulins is therefore likely to introduce errors into the partitioning of such sequences. PMID- 15345031 TI - Performance evaluation of commercial short-oligonucleotide microarrays and the impact of noise in making cross-platform correlations. AB - BACKGROUND: Despite the widespread use of microarrays, much ambiguity regarding data analysis, interpretation and correlation of the different technologies exists. There is a considerable amount of interest in correlating results obtained between different microarray platforms. To date, only a few cross platform evaluations have been published and unfortunately, no guidelines have been established on the best methods of making such correlations. To address this issue we conducted a thorough evaluation of two commercial microarray platforms to determine an appropriate methodology for making cross-platform correlations. RESULTS: In this study, expression measurements for 10,763 genes uniquely represented on Affymetrix U133A/B GeneChips and Amersham CodeLink UniSet Human 20 K microarrays were compared. For each microarray platform, five technical replicates, derived from the same total RNA samples, were labeled, hybridized, and quantified according to each manufacturers' standard protocols. The correlation coefficient (r) of differential expression ratios for the entire set of 10,763 overlapping genes was 0.62 between platforms. However, the correlation improved significantly (r = 0.79) when genes within noise were excluded. In addition to levels of inter-platform correlation, we evaluated precision, statistical-significance profiles, power, and noise levels for each microarray platform. Accuracy of differential expression was measured against real-time PCR for 25 genes and both platforms correlated well with r values of 0.92 and 0.79 for CodeLink and GeneChip, respectively. CONCLUSIONS: As a result of this study, we recommend using only genes called 'present' in cross-platform correlations. However, as in this study, a large number of genes may be lost from the correlation due to differing levels of noise between platforms. This is an important consideration given the apparent difference in sensitivity of the two platforms. Data from microarray analysis need to be interpreted cautiously and therefore, we provide guidelines for making cross-platform correlations. In all, this study represents the most comprehensive and specifically designed comparison of short-oligonucleotide microarray platforms to date using the largest set of overlapping genes. PMID- 15345032 TI - Automatic annotation of protein motif function with Gene Ontology terms. AB - BACKGROUND: Conserved protein sequence motifs are short stretches of amino acid sequence patterns that potentially encode the function of proteins. Several sequence pattern searching algorithms and programs exist foridentifying candidate protein motifs at the whole genome level. However, a much needed and important task is to determine the functions of the newly identified protein motifs. The Gene Ontology (GO) project is an endeavor to annotate the function of genes or protein sequences with terms from a dynamic, controlled vocabulary and these annotations serve well as a knowledge base. RESULTS: This paper presents methods to mine the GO knowledge base and use the association between the GO terms assigned to a sequence and the motifs matched by the same sequence as evidence for predicting the functions of novel protein motifs automatically. The task of assigning GO terms to protein motifs is viewed as both a binary classification and information retrieval problem, where PROSITE motifs are used as samples for mode training and functional prediction. The mutual information of a motif and aGO term association is found to be a very useful feature. We take advantage of the known motifs to train a logistic regression classifier, which allows us to combine mutual information with other frequency-based features and obtain a probability of correct association. The trained logistic regression model has intuitively meaningful and logically plausible parameter values, and performs very well empirically according to our evaluation criteria. CONCLUSIONS: In this research, different methods for automatic annotation of protein motifs have been investigated. Empirical result demonstrated that the methods have a great potential for detecting and augmenting information about the functions of newly discovered candidate protein motifs. PMID- 15345033 TI - SED, a normalization free method for DNA microarray data analysis. AB - BACKGROUND: Analysis of DNA microarray data usually begins with a normalization step where intensities of different arrays are adjusted to the same scale so that the intensity levels from different arrays can be compared with one other. Both simple total array intensity-based as well as more complex "local intensity level" dependent normalization methods have been developed, some of which are widely used. Much less developed methods for microarray data analysis include those that bypass the normalization step and therefore yield results that are not confounded by potential normalization errors. RESULTS: Instead of focusing on the raw intensity levels, we developed a new method for microarray data analysis that maps each gene's expression intensity level to a high dimensional space of SEDs (Signs of Expression Difference), the signs of the expression intensity difference between a given gene and every other gene on the array. Since SED are unchanged under any monotonic transformation of intensity levels, the SED based method is normalization free. When tested on a multi-class tumor classification problem, simple Naive Bayes and Nearest Neighbor methods using the SED approach gave results comparable with normalized intensity-based algorithms. Furthermore, a high percentage of classifiers based on a single gene's SED gave good classification results, suggesting that SED does capture essential information from the intensity levels. CONCLUSION: The results of testing this new method on multi-class tumor classification problems suggests that the SED-based, normalization-free method of microarray data analysis is feasible and promising. PMID- 15345034 TI - Making sense of centromeres. AB - Comparative analysis of the proteins that bind exclusively at the centromere provides evidence of an evolutionary battle that may make sense of sex. PMID- 15345035 TI - Adaptive evolution of centromere proteins in plants and animals. AB - BACKGROUND: Centromeres represent the last frontiers of plant and animal genomics. Although they perform a conserved function in chromosome segregation, centromeres are typically composed of repetitive satellite sequences that are rapidly evolving. The nucleosomes of centromeres are characterized by a special H3-like histone (CenH3), which evolves rapidly and adaptively in Drosophila and Arabidopsis. Most plant, animal and fungal centromeres also bind a large protein, centromere protein C (CENP-C), that is characterized by a single 24 amino-acid motif (CENPC motif). RESULTS: Whereas we find no evidence that mammalian CenH3 (CENP-A) has been evolving adaptively, mammalian CENP-C proteins contain adaptively evolving regions that overlap with regions of DNA-binding activity. In plants we find that CENP-C proteins have complex duplicated regions, with conserved amino and carboxyl termini that are dissimilar in sequence to their counterparts in animals and fungi. Comparisons of Cenpc genes from Arabidopsis species and from grasses revealed multiple regions that are under positive selection, including duplicated exons in some grasses. In contrast to plants and animals, yeast CENP-C (Mif2p) is under negative selection. CONCLUSIONS: CENP-Cs in all plant and animal lineages examined have regions that are rapidly and adaptively evolving. To explain these remarkable evolutionary features for a single-copy gene that is needed at every mitosis, we propose that CENP-Cs, like some CenH3s, suppress meiotic drive of centromeres during female meiosis. This process can account for the rapid evolution and the complexity of centromeric DNA in plants and animals as compared to fungi. PMID- 15345036 TI - The phosphatidylserine receptor has essential functions during embryogenesis but not in apoptotic cell removal. AB - BACKGROUND: Phagocytosis of apoptotic cells is fundamental to animal development, immune function and cellular homeostasis. The phosphatidylserine receptor (Ptdsr) on phagocytes has been implicated in the recognition and engulfment of apoptotic cells and in anti-inflammatory signaling. To determine the biological function of the phosphatidylserine receptor in vivo, we inactivated the Ptdsr gene in the mouse. RESULTS: Ablation of Ptdsr function in mice causes perinatal lethality, growth retardation and a delay in terminal differentiation of the kidney, intestine, liver and lungs during embryogenesis. Moreover, eye development can be severely disturbed, ranging from defects in retinal differentiation to complete unilateral or bilateral absence of eyes. Ptdsr -/- mice with anophthalmia develop novel lesions, with induction of ectopic retinal-pigmented epithelium in nasal cavities. A comprehensive investigation of apoptotic cell clearance in vivo and in vitro demonstrated that engulfment of apoptotic cells was normal in Ptdsr knockout mice, but Ptdsr-deficient macrophages were impaired in pro- and anti inflammatory cytokine signaling after stimulation with apoptotic cells or with lipopolysaccharide. CONCLUSION: Ptdsr is essential for the development and differentiation of multiple organs during embryogenesis but not for apoptotic cell removal. Ptdsr may thus have a novel, unexpected developmental function as an important differentiation-promoting gene. Moreover, Ptdsr is not required for apoptotic cell clearance by macrophages but seems to be necessary for the regulation of macrophage cytokine responses. These results clearly contradict the current view that the phosphatidylserine receptor primarily functions in apoptotic cell clearance. PMID- 15345037 TI - Fame is a bubble, but not for some. AB - Brilliant, acerbic, not given to suffer much of anybody gladly, let alone fools, Francis Crick had enormous influence that was due to his style and high scientific standards. PMID- 15345038 TI - Predicting and validating microRNA targets. AB - Given that microRNAs select their targets by nucleotide base-pairing, it follows that it should be possible to find microRNA targets computationally. There has been considerable progress, but assessing success and biological significance requires a move into the 'wet' lab. PMID- 15345039 TI - What's in a centromere? AB - The complete sequence of rice centromere 8 reveals a small amount of centromere specific satellite sequence in blocks interrupted by retrotransposons and other repetitive DNA, in an arrangement that is strikingly similar in overall size and content to other centromeres of multicellular eukaryotes. PMID- 15345040 TI - Chemical genomics in yeast. AB - Many drugs have unknown, controversial or multiple mechanisms of action. Four recent 'chemical genomic' studies, using genome-scale collections of yeast gene deletions that were either arrayed or barcoded, have presented complementary approaches to identifying gene-drug and pathway-drug interactions. PMID- 15345041 TI - The cyclooxygenases. AB - Cyclooxygenases (COXs) catalyze the rate-limiting step in the production of prostaglandins, bioactive compounds involved in processes such as fever and sensitivity to pain, and are the target of aspirin-like drugs. COX genes have been cloned from coral, tunicates and vertebrates, and in all the phyla where they are found, there are two genes encoding two COX isoenzymes; it is unclear whether these genes arose from an early single duplication event or from multiple independent duplications in evolution. The intron-exon arrangement of COX genes is completely conserved in vertebrates and mostly conserved in all species. Exon boundaries largely define the four functional domains of the encoded protein: the amino-terminal hydrophobic signal peptide, the dimerization domain, the membrane binding domain, and the catalytic domain. The catalytic domain of each enzyme contains distinct peroxidase and cyclooxygenase active sites; COXs are classified as members of the myeloperoxidase family. All COXs are homodimers and monotopic membrane proteins (inserted into only one leaflet of the membrane), and they appear to be targeted to the lumenal membrane of the endoplasmic reticulum, where they are N-glycosylated. In mammals, the two COX genes encode a constitutive isoenzyme (COX-1) and an inducible isoenzyme (COX-2); both are of significant pharmacological importance. PMID- 15345042 TI - RNAi for research and therapy. PMID- 15345043 TI - Structural genomics and structural biology: compare and contrast. PMID- 15345044 TI - Celebrating 40 years of biochemistry in Europe. PMID- 15345045 TI - Computational identification of developmental enhancers: conservation and function of transcription factor binding-site clusters in Drosophila melanogaster and Drosophila pseudoobscura. AB - BACKGROUND: The identification of sequences that control transcription in metazoans is a major goal of genome analysis. In a previous study, we demonstrated that searching for clusters of predicted transcription factor binding sites could discover active regulatory sequences, and identified 37 regions of the Drosophila melanogaster genome with high densities of predicted binding sites for five transcription factors involved in anterior-posterior embryonic patterning. Nine of these clusters overlapped known enhancers. Here, we report the results of in vivo functional analysis of 27 remaining clusters. RESULTS: We generated transgenic flies carrying each cluster attached to a basal promoter and reporter gene, and assayed embryos for reporter gene expression. Six clusters are enhancers of adjacent genes: giant, fushi tarazu, odd-skipped, nubbin, squeeze and pdm2; three drive expression in patterns unrelated to those of neighboring genes; the remaining 18 do not appear to have enhancer activity. We used the Drosophila pseudoobscura genome to compare patterns of evolution in and around the 15 positive and 18 false-positive predictions. Although conservation of primary sequence cannot distinguish true from false positives, conservation of binding-site clustering accurately discriminates functional binding-site clusters from those with no function. We incorporated conservation of binding-site clustering into a new genome-wide enhancer screen, and predict several hundred new regulatory sequences, including 85 adjacent to genes with embryonic patterns. CONCLUSIONS: Measuring conservation of sequence features closely linked to function--such as binding-site clusterin--makes better use of comparative sequence data than commonly used methods that examine only sequence identity. PMID- 15345046 TI - Global nucleosome occupancy in yeast. AB - BACKGROUND: Although eukaryotic genomes are generally thought to be entirely chromatin-associated, the activated PHO5 promoter in yeast is largely devoid of nucleosomes. We systematically evaluated nucleosome occupancy in yeast promoters by immunoprecipitating nucleosomal DNA and quantifying enrichment by microarrays. RESULTS: Nucleosome depletion is observed in promoters that regulate active genes and/or contain multiple evolutionarily conserved motifs that recruit transcription factors. The Rap1 consensus was the only binding motif identified in a completely unbiased search of nucleosome-depleted promoters. Nucleosome depletion in the vicinity of Rap1 consensus sites in ribosomal protein gene promoters was also observed by real-time PCR and micrococcal nuclease digestion. Nucleosome occupancy in these regions was increased by the small molecule rapamycin or, in the case of the RPS11B promoter, by removing the Rap1 consensus sites. CONCLUSIONS: The presence of transcription factor-binding motifs is an important determinant of nucleosome depletion. Most motifs are associated with marked depletion only when they appear in combination, consistent with a model in which transcription factors act collaboratively to exclude nucleosomes and gain access to target sites in the DNA. In contrast, Rap1-binding sites cause marked depletion under steady-state conditions. We speculate that nucleosome depletion enables Rap1 to define chromatin domains and alter them in response to environmental cues. PMID- 15345047 TI - A first-draft human protein-interaction map. AB - BACKGROUND: Protein-interaction maps are powerful tools for suggesting the cellular functions of genes. Although large-scale protein-interaction maps have been generated for several invertebrate species, projects of a similar scale have not yet been described for any mammal. Because many physical interactions are conserved between species, it should be possible to infer information about human protein interactions (and hence protein function) using model organism protein interaction datasets. RESULTS: Here we describe a network of over 70,000 predicted physical interactions between around 6,200 human proteins generated using the data from lower eukaryotic protein-interaction maps. The physiological relevance of this network is supported by its ability to preferentially connect human proteins that share the same functional annotations, and we show how the network can be used to successfully predict the functions of human proteins. We find that combining interaction datasets from a single organism (but generated using independent assays) and combining interaction datasets from two organisms (but generated using the same assay) are both very effective ways of further improving the accuracy of protein-interaction maps. CONCLUSIONS: The complete network predicts interactions for a third of human genes, including 448 human disease genes and 1,482 genes of unknown function, and so provides a rich framework for biomedical research. PMID- 15345048 TI - Comprehensive analysis of pseudogenes in prokaryotes: widespread gene decay and failure of putative horizontally transferred genes. AB - BACKGROUND: Pseudogenes often manifest themselves as disabled copies of known genes. In prokaryotes, it was generally believed (with a few well-known exceptions) that they were rare. RESULTS: We have carried out a comprehensive analysis of the occurrence of pseudogenes in a diverse selection of 64 prokaryote genomes. Overall, we find a total of around 7,000 candidate pseudogenes. Moreover, in all the genomes surveyed, pseudogenes occur in at least 1 to 5% of all gene-like sequences, with some genomes having considerably higher occurrence. Although many large populations of pseudogenes arise from large, diverse protein families (for example, the ABC transporters), notable numbers of pseudogenes are associated with specific families that do not occur that widely. These include the cytochrome P450 and PPE families (PF00067 and PF00823) and others that have a direct role in DNA transposition. CONCLUSIONS: We find suggestive evidence that a large fraction of prokaryote pseudogenes arose from failed horizontal transfer events. In particular, we find that pseudogenes are more than twice as likely as genes to have anomalous codon usage associated with horizontal transfer. Moreover, we found a significant difference in the number of horizontally transferred pseudogenes in pathogenic and non-pathogenic strains of Escherichia coli. PMID- 15345049 TI - Prediction and identification of Arabidopsis thaliana microRNAs and their mRNA targets. AB - BACKGROUND: A class of eukaryotic non-coding RNAs termed microRNAs (miRNAs) interact with target mRNAs by sequence complementarity to regulate their expression. The low abundance of some miRNAs and their time- and tissue-specific expression patterns make experimental miRNA identification difficult. We present here a computational method for genome-wide prediction of Arabidopsis thaliana microRNAs and their target mRNAs. This method uses characteristic features of known plant miRNAs as criteria to search for miRNAs conserved between Arabidopsis and Oryza sativa. Extensive sequence complementarity between miRNAs and their target mRNAs is used to predict miRNA-regulated Arabidopsis transcripts. RESULTS: Our prediction covered 63% of known Arabidopsis miRNAs and identified 83 new miRNAs. Evidence for the expression of 25 predicted miRNAs came from northern blots, their presence in the Arabidopsis Small RNA Project database, and massively parallel signature sequencing (MPSS) data. Putative targets functionally conserved between Arabidopsis and O. sativa were identified for most newly identified miRNAs. Independent microarray data showed that the expression levels of some mRNA targets anti-correlated with the accumulation pattern of their corresponding regulatory miRNAs. The cleavage of three target mRNAs by miRNA binding was validated in 5' RACE experiments. CONCLUSIONS: We identified new plant miRNAs conserved between Arabidopsis and O. sativa and report a wide range of transcripts as potential miRNA targets. Because MPSS data are generated from polyadenylated RNA molecules, our results suggest that at least some miRNA precursors are polyadenylated at certain stages. The broad range of putative miRNA targets indicates that miRNAs participate in the regulation of a variety of biological processes. PMID- 15345050 TI - Discovery of estrogen receptor alpha target genes and response elements in breast tumor cells. AB - BACKGROUND: Estrogens and their receptors are important in human development, physiology and disease. In this study, we utilized an integrated genome-wide molecular and computational approach to characterize the interaction between the activated estrogen receptor (ER) and the regulatory elements of candidate target genes. RESULTS: Of around 19,000 genes surveyed in this study, we observed 137 ER regulated genes in T-47D cells, of which only 89 were direct target genes. Meta analysis of heterogeneous in vitro and in vivo datasets showed that the expression profiles in T-47D and MCF-7 cells are remarkably similar and overlap with genes differentially expressed between ER-positive and ER-negative tumors. Computational analysis revealed a significant enrichment of putative estrogen response elements (EREs) in the cis-regulatory regions of direct target genes. Chromatin immunoprecipitation confirmed ligand-dependent ER binding at the computationally predicted EREs in our highest ranked ER direct target genes, NRIP1, GREB1 and ABCA3. Wider examination of the cis-regulatory regions flanking the transcriptional start sites showed species conservation in mouse-human comparisons in only 6% of predicted EREs. CONCLUSIONS: Only a small core set of human genes, validated across experimental systems and closely associated with ER status in breast tumors, appear to be sufficient to induce ER effects in breast cancer cells. That cis-regulatory regions of these core ER target genes are poorly conserved suggests that different evolutionary mechanisms are operative at transcriptional control elements than at coding regions. These results predict that certain biological effects of estrogen signaling will differ between mouse and human to a larger extent than previously thought. PMID- 15345051 TI - An Ambystoma mexicanum EST sequencing project: analysis of 17,352 expressed sequence tags from embryonic and regenerating blastema cDNA libraries. AB - BACKGROUND: The ambystomatid salamander, Ambystoma mexicanum (axolotl), is an important model organism in evolutionary and regeneration research but relatively little sequence information has so far been available. This is a major limitation for molecular studies on caudate development, regeneration and evolution. To address this lack of sequence information we have generated an expressed sequence tag (EST) database for A. mexicanum. RESULTS: Two cDNA libraries, one made from stage 18-22 embryos and the other from day-6 regenerating tail blastemas, generated 17,352 sequences. From the sequenced ESTs, 6,377 contigs were assembled that probably represent 25% of the expressed genes in this organism. Sequence comparison revealed significant homology to entries in the NCBI non-redundant database. Further examination of this gene set revealed the presence of genes involved in important cell and developmental processes, including cell proliferation, cell differentiation and cell-cell communication. On the basis of these data, we have performed phylogenetic analysis of key cell-cycle regulators. Interestingly, while cell-cycle proteins such as the cyclin B family display expected evolutionary relationships, the cyclin-dependent kinase inhibitor 1 gene family shows an unusual evolutionary behavior among the amphibians. CONCLUSIONS: Our analysis reveals the importance of a comprehensive sequence set from a representative of the Caudata and illustrates that the EST sequence database is a rich source of molecular, developmental and regeneration studies. To aid in data mining, the ESTs have been organized into an easily searchable database that is freely available online. PMID- 15345052 TI - Microarray analysis of microRNA expression in the developing mammalian brain. AB - BACKGROUND: MicroRNAs are a large new class of tiny regulatory RNAs found in nematodes, plants, insects and mammals. MicroRNAs are thought to act as post transcriptional modulators of gene expression. In invertebrates microRNAs have been implicated as regulators of developmental timing, neuronal differentiation, cell proliferation, programmed cell death and fat metabolism. Little is known about the roles of microRNAs in mammals. RESULTS: We isolated 18-26 nucleotide RNAs from developing rat and monkey brains. From the sequences of these RNAs and the sequences of the rat and human genomes we determined which of these small RNAs are likely to have derived from stem-loop precursors typical of microRNAs. Next, we developed a microarray technology suitable for detecting microRNAs and printed a microRNA microarray representing 138 mammalian microRNAs corresponding to the sequences of the microRNAs we cloned as well as to other known microRNAs. We used this microarray to determine the profile of microRNAs expressed in the developing mouse brain. We observed a temporal wave of expression of microRNAs, suggesting that microRNAs play important roles in the development of the mammalian brain. CONCLUSION: We describe a microarray technology that can be used to analyze the expression of microRNAs and of other small RNAs. MicroRNA microarrays offer a new tool that should facilitate studies of the biological roles of microRNAs. We used this method to determine the microRNA expression profile during mouse brain development and observed a temporal wave of gene expression of sequential classes of microRNAs. PMID- 15345054 TI - Improving identification of differentially expressed genes in microarray studies using information from public databases. AB - We demonstrate that the process of identifying differentially expressed genes in microarray studies with small sample sizes can be substantially improved by extracting information from a large number of datasets accumulated in public databases. The improvement comes from more reliable estimates of gene-specific variances based on other datasets. For a two-group comparison with two arrays in each group, for example, the result of our method was comparable to that of a t test analysis with five samples in each group or to that of a regularized t-test analysis with three samples in each group. Our results are further improved by weighting the results of our approach with the regularized t-test results in a hybrid method. PMID- 15345053 TI - Function-informed transcriptome analysis of Drosophila renal tubule. AB - BACKGROUND: Comprehensive, tissue-specific, microarray analysis is a potent tool for the identification of tightly defined expression patterns that might be missed in whole-organism scans. We applied such an analysis to Drosophila melanogaster Malpighian (renal) tubule, a defined differentiated tissue. RESULTS: The transcriptome of the D. melanogaster Malpighian tubule is highly reproducible and significantly different from that obtained from whole-organism arrays. More than 200 genes are more than 10-fold enriched and over 1,000 are significantly enriched. Of the top 200 genes, only 18 have previously been named, and only 45% have even estimates of function. In addition, 30 transcription factors, not previously implicated in tubule development, are shown to be enriched in adult tubule, and their expression patterns respect precisely the domains and cell types previously identified by enhancer trapping. Of Drosophila genes with close human disease homologs, 50 are enriched threefold or more, and eight enriched 10 fold or more, in tubule. Intriguingly, several of these diseases have human renal phenotypes, implying close conservation of renal function across 400 million years of divergent evolution. CONCLUSIONS: From those genes that are identifiable, a radically new view of the function of the tubule, emphasizing solute transport rather than fluid secretion, can be obtained. The results illustrate the phenotype gap: historically, the effort expended on a model organism has tended to concentrate on a relatively small set of processes, rather than on the spread of genes in the genome. PMID- 15345055 TI - A catalog of human cDNA expression clones and its application to structural genomics. AB - We describe here a systematic approach to the identification of human proteins and protein fragments that can be expressed as soluble proteins in Escherichia coli. A cDNA expression library of 10,825 clones was screened by small-scale expression and purification and 2,746 clones were identified. Sequence and protein-expression data were entered into a public database. A set of 163 clones was selected for structural analysis and 17 proteins were prepared for crystallization, leading to three new structures. PMID- 15345056 TI - Large-scale exploration of growth inhibition caused by overexpression of genomic fragments in Saccharomyces cerevisiae. AB - We have screened the genome of Saccharomyces cerevisiae for fragments that confer a growth-retardation phenotype when overexpressed in a multicopy plasmid with a tetracycline-regulatable (Tet-off) promoter. We selected 714 such fragments with a mean size of 700 base-pairs out of around 84,000 clones tested. These include 493 in-frame open reading frame fragments corresponding to 454 distinct genes (of which 91 are of unknown function), and 162 out-of-frame, antisense and intergenic genomic fragments, representing the largest collection of toxic inserts published so far in yeast. PMID- 15345057 TI - Is HIV-1 RNA dimerization a prerequisite for packaging? Yes, no, probably? AB - During virus assembly, all retroviruses specifically encapsidate two copies of full-length viral genomic RNA in the form of a non-covalently linked RNA dimer. The absolute conservation of this unique genome structure within the Retroviridae family is strong evidence that a dimerized genome is of critical importance to the viral life cycle. An obvious hypothesis is that retroviruses have evolved to preferentially package two copies of genomic RNA, and that dimerization ensures the proper packaging specificity for such a genome. However, this implies that dimerization must be a prerequisite for genome encapsidation, a notion that has been debated for many years. In this article, we review retroviral RNA dimerization and packaging, highlighting the research that has attempted to dissect the intricate relationship between these two processes in the context of HIV-1, and discuss the therapeutic potential of these putative antiretroviral targets. PMID- 15345058 TI - Emergence of new Salmonella Enteritidis phage types in Europe? Surveillance of infections in returning travellers. AB - BACKGROUND: Among human Salmonella Enteritidis infections, phage type 4 has been the dominant phage type in most countries in Western Europe during the last years. This is reflected in Salmonella infections among Swedish travellers returning from abroad. However, there are differences in phage type distribution between the countries, and this has also changed over time. METHODS: We used data from the Swedish infectious disease register and the national reference laboratory to describe phage type distribution of Salmonella Enteritidis infections in Swedish travellers from 1997 to 2002, and have compared this with national studies conducted in the countries visited. RESULTS: Infections among Swedish travellers correlate well with national studies conducted in the countries visited. In 2001 a change in phage type distribution in S. Enteritidis infections among Swedish travellers returning from some countries in southern Europe was observed, and a previously rare phage type (PT 14b) became one of the most commonly diagnosed that year, continuing into 2002 and 2003. CONCLUSIONS: Surveillance of infections among returning travellers can be helpful in detecting emerging infections and outbreaks in tourist destinations. The information needs to be communicated rapidly to all affected countries in order to expedite the implementation of appropriate investigations and preventive measures. PMID- 15345059 TI - Disability weights for the burden of oral disease in South Australia. AB - BACKGROUND: Australian burden of disease estimates appeared inconsistent with the reported repetitive and ubiquitous nature of dental problems. The aims of the study were to measure the nature, severity and duration of symptoms for specific oral conditions, and calculate disability weights from these measures. METHODS: Data were collected in 2001-02 from a random sample of South Australian dentists using mailed self-complete questionnaires. Dentists recorded the diagnosis of dental problems and provided patients with self-complete questionnaires to record the nature, severity and duration of symptoms using the EuroQol instrument. Data were available from 378 dentists (response rate = 60%). RESULTS: Disability weights were highest for pulpal infection (0.069), caries (0.044) and dentinal sensitivity (0.040), followed by denture problems (0.026), periodontal disease (0.023), failed restorations (0.019), tooth fractures (0.014) and tooth wear (0.011). Aesthetic problems had a low disability weight (0.002), and both recall/maintenance care and oral hygiene had adjusted weights of zero. CONCLUSIONS: Disability weights for caries (0.044), periodontal disease (0.023) and denture problems (0.026) in this study were higher than comparable oral health conditions in the Australian Burden of Disease and Injury Study (0.005 for caries involving a filling and 0.014 for caries involving an extraction, 0.007 for periodontal disease, and 0.004 for edentulism). A range of common problems such as pulpal infection, failed restorations and tooth fracture that were not included in the Australian Burden of Disease and Injury Study had relatively high disability weights. The inclusion of a fuller range of oral health problems along with revised disability weights would result in oral health accounting for a larger amount of disability than originally estimated. PMID- 15345060 TI - Immature rats show ovulatory defects similar to those in adult rats lacking prostaglandin and progesterone actions. AB - Gonadotropin-primed immature rats (GPIR) constitute a widely used model for the study of ovulation. Although the equivalence between the ovulatory process in immature and adult rats is generally assumed, the morphological and functional characteristics of ovulation in immature rats have been scarcely considered. We describe herein the morphological aspects of the ovulatory process in GPIR and their response to classical ovulation inhibitors, such as the inhibitor of prostaglandin (PG) synthesis indomethacin (INDO) and a progesterone (P) receptor (PR) antagonist (RU486). Immature Wistar rats were primed with equine chorionic gonadotropin (eCG) at 21, 23 or 25 days of age, injected with human chorionic gonadotropin (hCG) 48 h later, and sacrificed 16 h after hCG treatment, to assess follicle rupture and ovulation. Surprisingly, GPIR showed age-related ovulatory defects close similar to those in adult rats lacking P and PG actions. Rats primed with eCG at 21 or 23 days of age showed abnormally ruptured corpora lutea in which the cumulus-oocyte complex (COC) was trapped or had been released to the ovarian interstitum, invading the ovarian stroma and blood and lymphatic vessels. Supplementation of immature rats with exogenous P and/or PG of the E series did not significantly inhibit abnormal follicle rupture. Otherwise, ovulatory defects were practically absent in rats primed with eCG at 25 days of age. GPIR treated with INDO showed the same ovulatory alterations than vehicle-treated ones, although affecting to a higher proportion of follicles. Blocking P actions with RU486 increased the number of COC trapped inside corpora lutea and decreased ovulation. The presence of ovulatory defects in GPIR, suggests that the capacity of the immature ovary to undergo the coordinate changes leading to effective ovulation is not fully established in Wistar rats primed with eCG before 25 days of age. PMID- 15345061 TI - 17beta-estradiol induced vitellogenesis is inhibited by cortisol at the post transcriptional level in Arctic char (Salvelinus alpinus). AB - This study was performed to investigate stress effects on the synthesis of egg yolk precursor, vitellogenin (Vtg) in Arctic char (Salvelinus alpinus). In particular the effect of cortisol (F) was determined since this stress hormone has been suggested to interfere with vitellogenesis and is upregulated during sexual maturation in teleosts. Arctic char Vtg was purified and polyclonal antibodies were produced in order to develop tools to study regulation of vitellogenesis. The Vtg antibodies were used to develop an enzyme-linked immunosorbent assay. The corresponding Vtg cDNA was cloned from a hepatic cDNA library in order to obtain DNA probes to measure Vtg mRNA expression. Analysis of plasma from juvenile Arctic char, of both sexes, exposed to different steroids showed that production of Vtg was induced in a dose dependent fashion by 17beta estradiol (E2), estrone and estriol. Apart from estrogens a high dose of F also upregulated Vtg. In addition, F, progesterone (P) and tamoxifen were tested to determine these compounds ability to modulate E2 induced Vtg synthesis at both the mRNA and protein level. Tamoxifen was found to inhibit E2 induced Vtg mRNA and protein upregulation. P did not alter the Vtg induction while F reduced the Vtg protein levels without affecting the Vtg mRNA levels. Furthermore the inhibition of Vtg protein was found to be dose dependent. Thus, the inhibitory effect of F on Vtg appears to be mediated at the post-transcriptional level. PMID- 15345062 TI - The Menopause Rating Scale (MRS) scale: a methodological review. AB - BACKGROUND: This paper compiles data from different sources to get a first comprehensive picture of psychometric and other methodological characteristics of the Menopause Rating Scale (MRS) scale. The scale was designed and standardized as a self-administered scale to (a) to assess symptoms/complaints of aging women under different conditions, (b) to evaluate the severity of symptoms over time, and (c) to measure changes pre- and postmenopause replacement therapy. The scale became widespread used (available in 10 languages). METHOD: A large multinational survey (9 countries in 4 continents) from 2001/ 2002 is the basis for in depth analyses on reliability and validity of the MRS. Additional small convenience samples were used to get first impressions about test-retest reliability. The data were centrally analyzed. Data from a postmarketing HRT study were used to estimate discriminative validity. RESULTS: Reliability measures (consistency and test-retest stability) were found to be good across countries, although the sample size for test-retest reliability was small. VALIDITY: The internal structure of the MRS across countries was astonishingly similar to conclude that the scale really measures the same phenomenon in symptomatic women. The sub scores and total score correlations were high (0.7-0.9) but lower among the sub scales (0.5-0.7). This however suggests that the subscales are not fully independent. Norm values from different populations were presented showing that a direct comparison between Europe and North America is possible, but caution recommended with comparisons of data from Latin America and Indonesia. But this will not affect intra-individual comparisons within clinical trials. The comparison with the Kupperman Index showed sufficiently good correlations, illustrating an adept criterion-oriented validity. The same is true for the comparison with the generic quality-of-life scale SF-36 where also a sufficiently close association has been shown. CONCLUSION: The currently available methodological evidence points towards a high quality of the MRS scale to measure and to compare HRQoL of aging women in different regions and over time, it suggests a high reliability and high validity as far as the process of construct validation could be completed yet. PMID- 15345064 TI - Women's Health Surveillance Report: Introduction. PMID- 15345063 TI - Strategies for the recovery of active proteins through refolding of bacterial inclusion body proteins. AB - Recent advances in generating active proteins through refolding of bacterial inclusion body proteins are summarized in conjunction with a short overview on inclusion body isolation and solubilization procedures. In particular, the pros and cons of well-established robust refolding techniques such as direct dilution as well as less common ones such as diafiltration or chromatographic processes including size exclusion chromatography, matrix- or affinity-based techniques and hydrophobic interaction chromatography are discussed. Moreover, the effect of physical variables (temperature and pressure) as well as the presence of buffer additives on the refolding process is elucidated. In particular, the impact of protein stabilizing or destabilizing low- and high-molecular weight additives as well as micellar and liposomal systems on protein refolding is illustrated. Also, techniques mimicking the principles encountered during in vivo folding such as processes based on natural and artificial chaperones and propeptide-assisted protein refolding are presented. Moreover, the special requirements for the generation of disulfide bonded proteins and the specific problems and solutions, which arise during process integration are discussed. Finally, the different strategies are examined regarding their applicability for large-scale production processes or high-throughput screening procedures. PMID- 15345065 TI - The Social Context of Women's Health. AB - HEALTH ISSUE: The discussion of health emphasizes the importance of analyses of social determinants of health. Social determinants permit the targeting of policies towards the social factors that impair or improve health. Two broad questions are considered: (i) what do we know about the social determinants of women's health? (ii) are there gender-related differences in health problems, and how we might explain them? KEY FINDINGS: While 'sex' may be used to denote the biological difference between women and men, it is an imperfect measure of 'gender'. It is argued that a single measure cannot hope to capture the complexity of gender nor the ways in which gender relations change over time and give rise to or exacerbate health problems. The literature on the social determinants of health shows the importance of placing a primary emphasis on addressing the social and economic sources of ill health at national, provincial and community levels. DATA GAPS AND RECOMMENDATIONS: Recent studies of gender differences in health point to a lack of data and to the importance of understanding changing gender relations; differences in power and access to resources between women and men, and changing expectations of appropriate gender roles and behaviours. Poverty, social exclusion, unemployment, poor working conditions and unequal gender relations have a profound influence on patterns of health and illness. We suggest some material markers of change that might be used in health surveillance. With a more complete understanding of gender's role in shaping daily lives, these markers could be refined and expanded. PMID- 15345066 TI - Multiple Roles and Women's Mental Health in Canada. AB - HEALTH ISSUE: Research on the relationship between women's social roles and mental health has been equivocal. Although a greater number of roles often protect mental health, certain combinations can lead to strain. Our study explored the moderating affects of different role combinations on women's mental health by examining associations with socioeconomic status and differences in women's distress (depressive symptoms, personal stress (role strain) and chronic stress (role strain plus environmental stressors). KEY FINDINGS: Women with children, whether single or partnered, had a higher risk of personal stress. Distress, stress and chronic stress levels of mothers, regardless of employment, or marital status, are staggeringly high. Single, unemployed mothers were significantly more likely than all other groups to experience financial stress and food insecurity. For partnered mothers, rates of personal stress and chronic stress were significantly lower among unemployed partnered mothers. Married and partnered mothers reported better mental health than their single counterparts. Lone, unemployed mothers were twice as likely to report a high level of distress compared with other groups. Lone mothers, regardless of employment status, were more likely to report high personal and chronic stress. DATA GAPS AND RECOMMENDATIONS: National health surveys need to collect more data on the characteristics of women's work environment and their care giving responsibilities. Questions on household composition should include inter generational households, same sex couples and multifamily arrangements. Data disaggregation by ethno-racial background would be helpful. Data should be collected on perceived quality of domestic and partnership roles and division of labours. PMID- 15345067 TI - Personal Health Practices. AB - HEALTH ISSUE: There are differences in health practices and self-rated health among different socio-demographic groups of women. The relationship between socio demographic status and a) a range of health behaviours and b) a combination of multiple risk and multiple health promoting practices were examined. The relationship between self-rated health and health practices was also assessed. KEY FINDINGS: There were geographic differences in health practices with women in British Columbia having the highest odds of engaging in multiple health promoting practices, while women in Quebec had the lowest. Reports of engaging in multiple risk behaviours were most common in Ontario. Women from Ontario had the highest odds of reporting very good/excellent health and women from British Columbia had among the lowest odds.The data supported a strong social gradient between an increase in income/education and healthy practices, especially those that are health promoting. However, women with higher education were more likely to be overweight and those with higher incomes were more likely to drink alcohol regularly.Immigrant women were less likely to engage in multiple health risk practices compared to Canadian-born women. However, they were less likely to report very good/ excellent health than non- immigrants. While marriage appeared to have a generally protective effect on women's health practices, single women were more likely to be physically active and have a normal weight. DATA GAPS AND RECOMMENDATIONS: More sensitive indicators need to be developed to better understand possible reasons for the socioeconomic gradient. Data collection should focus on both rural and Aboriginal populations. PMID- 15345068 TI - Body Weight and Body Image. AB - HEALTH ISSUE: Body weight is of physical and psychological importance to Canadian women; it is associated with health status, physical activity, body image, and self-esteem. Although the problems associated with overweight and obesity are indeed serious, there are also problems connected to being underweight. Weight prejudice and the dieting industry intensify body image concerns for Canadian women and can have a major negative impact on self-esteem. KEY FINDINGS: Women have lower BMIs than men, a lower incidence of being overweight and a higher incidence of being underweight. However, women across all weight categories are more dissatisfied with their bodies. Sixty percent of women are inactive, and women with a BMI of 27 or higher are more likely to be inactive than women with lower BMIs. The data show that women are aware of the health benefits of exercise, but there is a gap between knowledge and practice. When asked about barriers to health improvement, 39.7% of women cited lack of time and 39.2% lack of willpower. DATA GAPS AND RECOMMENDATIONS: Weight prejudice must be made unacceptable and positive body image should be encouraged and diversity valued. Health policies should encourage healthy eating and healthy activity. Health curricula for young students should include information about healthy eating, active lifestyle, and self-esteem. Physical activities that mothers can participate in with their families should be encouraged. Research should be funded to elucidate the most effective methods of getting women to become and remain physically active without focusing on appearance. PMID- 15345069 TI - Physical Activity and Obesity in Canadian Women. AB - HEALTH ISSUE: Overweight and obesity have been recognized as major public health concern in Canada and throughout the world. Lack of physical activity, through its impact on energy balance, has been identified as an important modifiable risk factor for obesity. Physical activity and obesity are also important risk factors for a variety of chronic diseases. This chapter provides an overview of the current state of physical activity and overweight/obesity among Canadian women. KEY FINDINGS: For all ages combined more women (57%) than men (50%) are physically inactive (energy expenditure <1.5 KKD). Physical activity increases as income adequacy and educational level decrease. Physical inactivity also varies by ethnicity. The prevalence of both overweight (BMI 25.0 - 29.9 kg/m2) and obese (BMI >/= 30 kg/m2) Canadian women has increased 7% since 1985. Obesity increases with age and is highest among women reporting low and lower middle incomes and lower levels of education. The prevalence of obesity is highest among Aboriginal women and men (28% and 22% respectively). DATA GAPS AND RECOMMENDATIONS: There is currently no surveillance system in Canada to monitor the level of physical activity among children, those performing activity at work, at school or in the home. There is a gap in the knowledge surrounding socio-cultural and ecological determinants of physical activity and obesity and the associations of these to chronic disease among women and minority populations. Multi-sectoral policy interventions that act to decrease the broad systemic barriers to physical activity and healthy weights among all women are needed. PMID- 15345070 TI - Gender Differences in Smoking and Self Reported Indicators of Health. AB - HEALTH ISSUE: Smoking among Canadian women is a serious public health issue. Using the 1998-99 National Population Health Survey, this study examined underlying factors contributing to differences in prevalence of smoking among subgroups of women and men, and its effects on self-reported indicators of health. KEY FINDINGS: In Canada, 26.4% of women and 29.2% of men were classified as current smokers. Higher levels of education and income were associated with decreased odds of current smoking. Adjusting for all other factors, being an ethnic minority decreased the odds of current smoking for both men and women (OR:0.35, 99%CI:0.23-0.54; OR:0.13, 99%CI: 0.09-0.20 respectively). Single mothers had the highest odds of smoking (OR: 2.12, 99%CI: 1.28-3.51) when compared to married mothers with children under 25 years of age. Current women smokers and current and former men smokers were less likely to report very good or excellent health compared with never smokers (OR: 0.83, 99%CI: 0.70-0.98; OR: 0.49, 99%CI: 0.41-0.60; OR: 0.75, 99%CI: 0.63-0.90 respectively). Women who were current smokers had increased odds of needing health care and not receiving it (OR: 1.50, 99%CI: 1.10-2.05). DATA GAPS AND RECOMMENDATIONS: Key issues for Canadian women include an increased prevalence of smoking among young girls and the strong association between smoking and social and economic disadvantage. Tobacco control policies and programs must target high-risk groups more effectively. Of particular importance is the development of programs and policies that do not serve to reinforce existing inequities, but rather, contribute to their amelioration. PMID- 15345071 TI - Women and Substance Abuse Problems. AB - HEALTH ISSUES: Differences exist in the prevalence and physical health impacts of problem substance use among men and women. These differences are also found in the mental health and trauma events related to substance use, barriers to treatment and harm-reduction services and the impact of substance use on pregnancy and parenting. Data from the 2000-2001 Canadian Community Health Survey and Canada's Alcohol and Other Drugs Survey (1994) were used to explore this issue further. KEY FINDINGS: While women use alcohol and illicit drugs at lower rates than men, the health impact of their use is significant, and in some cases greater than for men. Women are more likely to use prescribed psychoactive drugs (e.g. pain relievers, sleeping pills, tranquillizers) and most of these drugs have addictive potential and long-term negative consequences on health. Research collected from treatment centres in Canada show high rates of victimization experienced by women, which have implications for both their substance use treatment and improvement in mental health. DATA GAPS AND RECOMMENDATIONS: Significant gaps exist in our knowledge on the level, type, and impact of substance use and the adequacy of programming for Canadian women. Information that might be used to guide prevention initiatives, such as the amount of alcohol that might safely be used in pregnancy and the incidence of fetal alcohol syndrome, related birth defects and developmental disabilities are unknown. Improved surveillance, sensitive and comprehensive screening for substance use problems, accessible treatment and harm reduction programming, and coordination with the mental health and violence fields are recommended. PMID- 15345072 TI - Mortality: life and health expectancy of Canadian women. AB - HEALTH ISSUE: The sex differences in mortality, life expectancy, and, to a lesser extent, health expectancy, are well recognized in Canada and internationally. However, the factors explaining these differences between women and men are not well understood. This chapter explores the contribution of various causes of death (such as preventable, and sex-specific deaths) on these differences between women and men. KEY FINDINGS: "External" preventable causes of death (e.g. smoking related, injuries, etc.) were responsible for a large portion of the sex gap in mortality and life expectancy. When excluding these causes from the calculations, the sex gap in life expectancies were largely reduced, decreasing from approximately 5.5 years (life expectancy being 81.4, years in women, and 75.9 years in men) to approximately 2.2 years (84.9 in women and 82.7 in men). Sex gaps in corresponding health expectancies entirely disappeared when these preventable causes of death were excluded. Moreover, a larger death burden was observed among women than men for sex-specific causes of death (eg. excess breast cancer, gynaecological cancers, maternal mortality). Significant disparities were also observed in the mortality rates of various subgroups of women by geographic regions of Canada. DATA GAPS AND RECOMMENDATIONS: These results indicate that women do not appear to have a large biological survival advantage but, rather, are at lower risk of preventable deaths. They also provide additional information needed for the development of policies aimed at reducing disparities in life and health expectancies in Canada and other developed countries. PMID- 15345073 TI - Morbidity Experiences and Disability Among Canadian Women. AB - HEALTH ISSUE: Women are more frequently affected by chronic conditions and disability than men. Although some of these sex differences have been in part attributed to biological susceptibility, social determinants of health and other factors, these gaps have not been fully explained in the current literature. This chapter presents comparisons of hospitalization rates, and the prevalence of chronic conditions and physical disability between Canadian women and men and between various subgroups of women, adjusting for selected risk factors. The Canadian Hospital Morbidity Database (2000-2001) and Canadian Community Health Survey (2000-2001) were used to examine inpatient hospital morbidity, prevalence of chronic conditions and disability. KEY FINDINGS: Hospitalization rates were 20% higher among women than men. This was due to the large number of hospitalizations for pregnancies and childbirth. When "normal" deliveries were excluded, hospitalization rates remained higher among women. Women had slightly lower rates of hospitalizations for ambulatory-care sensitive conditions than men. Prevalence of activity limitation (mild and severe) was higher among women than men, and differences remained after adjusting for age, chronic conditions, socio-economic status, and smoking. Women who reported a disability were less likely than men to be in a partnered relationship, have less tangible social support, and have lower income and employment rates. DATA GAPS AND RECOMMENDATIONS: The impact of morbidity and disability on Canadian women is substantial. These results identify areas for interventions among more vulnerable subgroups, and point to the need for further research in the area of risk factors for the prevention of morbidity and disability in the population. PMID- 15345074 TI - The impact of a reduced fertility rate on women's health. AB - HEALTH ISSUE: Total fertility rates (TFRs) have decreased worldwide. The Canadian fertility rate has gone from 3.90 per woman in 1960 to 1.49 in 2000. However, not many studies have examined the impact on women's health of reduced fertility rates, delayed fertility and more births to unmarried women. This paper presents information on the relation between family size and specific determinants of health. KEY FINDINGS: The rate of TFR decline varies considerably by geographic location and socio-demographic subgroup. Further, the associations between family size and selected determinants of health are different for women and men. For example a woman with one child is almost four times more likely to be "coupled" than a childless woman, and if she has two children she is significantly more likely to be "coupled" than if she had only one child. However, a man with one or more children is over six times more likely to be "coupled" than his childless counterpart, and this does not vary with family size. DATA GAPS AND RECOMMENDATIONS: There is a paucity of data on the impact of reduced fertility rates on women's health in general and on how women's roles affect their decision to have children. While it would be useful to examine longer-term health outcomes by parity and age of first birth, as well as socio-economic and role-related variables these longitudinal and detailed "role related" data are not available. Given the differing profiles of women and men with children, further health policies research is needed to support vulnerable women with children. PMID- 15345075 TI - Breast Cancer in Canadian Women. AB - HEALTH ISSUE: Although lung cancer is the leading cause of cancer deaths for Canadian women, breast cancer is the most frequently diagnosed. About 5400 women are expected to die from this disease in 2003. In 1998, a woman's lifetime risk of breast cancer was about one in nine. KEY FINDINGS: A number of risk factors for breast cancer have been identified. These include advancing age, hormonal factors (eg. early menarche, late menopause and late age at first full-term pregnancy), familial risk, BRCA-1 and BRCA-2 gene mutations, diet and postmenopausal obesity.Several interventions have been introduced to assist women at high risk for breast cancer, including genetic counseling and testing for women who have strong family histories of breast cancer; selective estrogen receptor modifiers, such as tamoxifen, that has been shown to reduce breast cancer rates; prophylactic mastectomy and screening. DATA GAPS AND RECOMMENDATIONS: Guidelines are unclear in several areas, particularly in screening. Where clinical guidelines are available, health services research or ongoing monitoring (by provincial/territorial cancer agencies) is needed to assess compliance with the guidelines and to ensure equity of access within the provinces/territories.Key components of organized screening programs need to be established, in part to ensure that screening is carried out in high-quality, co ordinated programs. There is also a need to develop ways to involve women fully in informed decision-making and to address several policy issues to prevent disparities in access to high-quality services. Patenting issues associated with genetic tests also need to be clarified. PMID- 15345076 TI - Cancer of the Uterine Cervix. AB - HEALTH ISSUE: Cervical cancer is one of the most common malignant diseases of women; it is diagnosed in almost half a million women every year and half as many die from it annually. In Canada and other industrialized countries, its incidence has decreased due to cytology screening. However, invasive cases still occur, particularly among immigrant groups and native Canadian women. Although incidence of squamous cell carcinomas has decreased, the proportion of adenocarcinomas has increased because Pap cytology is ineffective to detect these lesions. KEY FINDINGS: In Canada, cervical cancer will cause an estimated 11,000 person-years of life lost. In most Canadian provinces, early detection is dependent on opportunistic screening. Primary prevention can be achieved through health education (sexual behavior modification) and vaccination to prevent infection from Human Papillomavirus (HPV). The initial results from vaccination trials are encouraging but wide scale use is more than a decade away. DATA GAPS AND RECOMMENDATIONS: Most cases of cervical cancer occur because the Pap smear was either false negative, was not done or not done often enough. Appropriate recommendations and guidelines exist on implementation of cytology-based programs. However, most Canadian women do not have access to organized screening. Further research is needed to 1) evaluate automated cytology systems; 2) define appropriate management of precursor lesions and 3) deliver definitive evidence of HPV testing efficacy in long-term follow-up studies with invasive cancer as an outcome and 4) provide Canadian data to justify augmenting or modifying current programs to use HPV testing in secondary triage of equivocal Pap smears. PMID- 15345077 TI - Other Gynecologic Cancers: endometrial, ovarian, vulvar and vaginal cancers. AB - HEALTH ISSUE: In Canada, cancers of the endometrium, ovaries, vulva, vagina, placenta and adnexa account for 11% of all malignant neoplasms in women and 81% of all genital cancers. Although the incidence and mortality from vulvar and vaginal cancers are very low, endometrium and ovarian cancer are important public health problems. KEY FINDINGS: In Canada, there has been no appreciable improvement in survival for women with advanced endometrial (EC) or ovarian cancer (OC) over the past 30 years. The prognosis of EC is good for most patients because diagnosis is made at early stages. However, survival of OC is poor; more than 70% of cases are diagnosed at late stages. Up to 10% of OCs is linked to familial aggregation. Cancers of the vulva and of the vagina are very rare. The survival experience for women with the latter is worse than for those with the former. Both share many risk factors with cervical cancer and the recent developments in the study of HPV infection should be applicable to these diseases as well. Of particular interest will be the advent of vaccines for the primary prevention of HPV infection. DATA GAPS AND RECOMMENDATIONS: At present, the best available means to diagnose gynecologic malignancies is a detailed clinical examination, considering the totality of information on potential and proven risk factors, such as age, reproductive health, sexual practices, use unopposed estrogens or of oral contraceptives or tubal ligation, obesity, diet, smoking, and the familial clustering of some of these cancers. PMID- 15345078 TI - Cardiovascular Disease. AB - HEALTH ISSUE: Cardiovascular disease (CVD) is the leading cause of death in Canadian women and men. In general, women present with a wider range of symptoms, are more likely to delay seeking medial care and are less likely to be investigated and treated with evidence-based medications, angioplasty or coronary artery bypass graft than men. KEY FINDINGS: In 1998, 78,964 Canadians died from CVD, almost half (39,197) were women. Acute myocardial infarction, which increases significantly after menopause, was the leading cause of death among women.Cardiovascular disease accounted for 21% of all hospital admissions for Canadian women over age 50 in 1999. Admissions to hospital for ischemic heart disease were more frequent for men, but the mean length of hospital stay was longer for women.Mean blood pressure increases with age in both men and women. After age 65, however, high blood pressure is more common among Canadian women. More than one-third of postmenopausal Canadian women have hypertension.Diabetes increases the mortality and morbidity associated with CVD in women more than it does in men. Depression also contributes to the incidence and recovery from CVD, particularly for women who experience twice the rate of depression as men. DATA GAPS AND RECOMMENDATIONS: CVD needs to be recognized as a woman's health issue given Canadian mortality projections (particularly heart failure). Health professionals should be trained to screen, track, and address CVD risk factors among women, including hypertension, elevated lipid levels, smoking, physical inactivity, depression, diabetes and low socio-economic status. PMID- 15345079 TI - Diabetes in Canadian Women. AB - HEALTH ISSUE: Diabetes mellitus (DM) is a chronic health condition affecting 4.8% of Canadian adults >/= 20 years of age. The prevalence increases with age. According to the National Diabetes Surveillance System (NDSS) (1998-1999), approximately 12% of Canadians aged 60-74 years are affected. One-third of cases may remain undiagnosed. The projected increase in DM prevalence largely results from rising rates of obesity and inactivity. KEY FINDINGS: DM in Canada appears to be more common among men than women. However, among Aboriginal Canadians, two thirds of affected individuals are women. Although obesity is more prevalent among men than women (35% vs. 27%), the DM risk associated with obesity is greater for women. Socio-economic status is inversely related to DM prevalence but the income-related disparities are greater among women. Polycystic ovarian syndrome affects 5-7% of reproductive-aged women and doubles their risk for DM. Women with gestational diabetes frequently develop DM over the next 10 years. DATA GAPS AND RECOMMENDATIONS: Studies of at risk ethnic/racial groups and women with gestational diabetes are needed. Age and culturally sensitive programs need to be developed and evaluated. Studies of low-income diabetic women are required before determining potential interventions. Lifestyle programs in schools and workplaces are needed to promote well-being and combat obesity/inactivity, together with lobbying of the food industry for needed changes. High depression rates among diabetic women influence self-care ability and health care expenditures. Health professionals need further training in the use of effective counseling skills that will assist people with DM to make and maintain difficult behavioural changes. PMID- 15345080 TI - Chronic Pain: The Extra Burden on Canadian Women. AB - HEALTH ISSUE: Chronic pain is a major health problem associated with significant costs to both afflicted individuals and society as a whole. These costs seem to be disproportionately borne by women, who generally have higher prevalence rates for chronic pain than do men. KEY FINDINGS: Data obtained from 125,574 respondents to the Canadian Community Health Survey (2000-2001) indicated that 18% of Canadian women suffered from chronic pain, compared to 14% of men. This gender discrepancy, however, seemed to be linked primarily to differences in age, income, and education between adult men and women in this large sample. Age, income, depression and functional interference with activities were strongly associated with chronic pain in general. No gender differences were found in the intensity of pain experienced. Ethnicity was not strongly associated with chronic pain prevalence, although Asians were the group with the highest chronic pain prevalence in the over-65 age group and Aboriginal Canadians had the highest prevalence in the under-65 age group. DATA GAPS AND RECOMMENDATIONS: Current gaps in our knowledge include the types of chronic pain women experience, their impact on domestic responsibilities and parenting and health care utilization patterns of women with chronic pain. Data sources such as provincial databases of billing claims may be useful in the future to enrich our knowledge of health care utilization and analgesic medication use. Enhanced surveillance, assessment, and early identification of pain disorders are recommended to improve outcomes. Considering current demographic patterns toward an older population, there is also some urgency to the development of patient education and self-management programs. PMID- 15345081 TI - The Impact of Arthritis on Canadian Women. AB - HEALTH ISSUE: Arthritis is one of the most prevalent chronic conditions in Canada and a leading cause of long-term disability, pain, and increased health care utilization. It is also a far more prevalent condition among women than men. Information was obtained primarily from the 1998-99 National Population Health Survey and the Canadian Joint Replacement Registry. KEY FINDINGS: In 1998, the overall prevalence of self-reported arthritis or rheumatism in Canadian women was 20.0%. This rate increased to 55.6% among women over 75 years of age. Compared to women with chronic conditions, women with arthritis were more likely to experience long-term disability; report worse health; experience more pain; be dependent upon others and consult general practitioners, specialists, and physiotherapists more frequently. While men and women with arthritis under utilize total joint replacement surgery, the degree of under-use was over three times greater for women. DATA GAPS AND RECOMMENDATIONS: There is a lack of detailed information on the use of health care services by women with arthritis. There are also no systematic data available on the prescribing of medications, access to services such as assistive devices or exercise programs, or use of community support, self-management strategies, or rehabilitation services. The burden of arthritis both on women and on society is expected to increase as the population ages. A comprehensive health strategy to reduce the impact of arthritis is required to ensure that health and support services are available in a timely manner and provided in such a way to meet the needs of Canadian women. PMID- 15345082 TI - Depression. AB - HEALTH ISSUE: Depression causes significant distress or impairment in physical, social, occupational and other key areas of functioning. Women are approximately twice as likely as men to experience depression. Psychosocial factors likely mediate the risks for depression incurred by biological influences. KEY FINDINGS: Data from the 1999 National Population Health Survey show that depression is more common among Canadian women, with an annual self-reported incidence of 5.7% compared with 2.9% in men. The highest rates of depression are seen among women of reproductive age. Predictive factors for depression include previous depression, feeling out of control or overwhelmed, chronic health problems, traumatic events in childhood or young adulthood, lack of emotional support, lone parenthood, and low sense of mastery. Although depression is treatable, only 43% of depressed women had consulted a health professional in 1998/99 and only 32.4% were taking antidepressant medication. People with lower education, inadequate income, and fewer contacts with a health professional were less likely to receive depression treatment. DATA GAPS AND RECOMMENDATIONS: A better understanding of factors that increase vulnerability and resilience to depression is needed. There is also a need for the collection and analysis of data pertaining to: prevalence of clinical anxiety; the prevalence of depression band 12 months after childbirth factors contributing to suicide contemplation and attempts among adolescent girls, current treatments for depression and their efficacy in depressed women at different life stages; interprovincial variation in depression rates and hospitalizations and the impact and costs of depression on work, family, individuals, and society. PMID- 15345083 TI - Dementia / Alzheimer's Disease. AB - HEALTH ISSUES: Dementia, including Alzheimer's disease (AD) increases exponentially with age from the age of 65. The number of people with dementia will increase significantly over the next three decades as the population ages. While prevalence and incidence rates do not differ markedly in women, compared to men, women live longer on average, so the number of women with dementia is greater than for men. Also, women are more frequently caregivers for people with dementia. Thus, dementia is an important health problem for women. KEY FINDINGS: The Canadian Study of Health and Aging showed an increase in prevalence of dementia with age for both sexes, approximately doubling every five years of age. Rates of AD were higher in women whereas rates of vascular dementia were higher in men. The risk of AD increased with increasing age, lower education, and apolipoprotein E epsilon4. Regular physical activity was clearly protective in women; this was less clear for men. Use of non-steroidal anti-inflammatory drugs, wine consumption, and past exposure to vaccines decreased the risk of AD. Estrogen replacement therapy did not reduce the risk of AD. About three quarters of caregivers for dementia patients were women. DATA GAPS AND RECOMMENDATIONS: The protective effect of regular physical activity for AD provides an additional reason to promote regular physical activity at all ages. Ongoing surveillance of the incidence, prevalence and risks for dementia is needed to monitor the impact of treatments as well as the aging of the population on the burden of dementia. PMID- 15345084 TI - Eating Disorders. AB - HEALTH ISSUE: Eating disorders are an increasing public health problem among young women. Anorexia and bulimia may give rise to serious physical conditions such as hypothermia, hypotension, electrolyte imbalance, endocrine disorders, and kidney failure. KEY ISSUES: Eating disorders are primarily a problem among women. In Ontario in 1995, over 90% of reported hospitalized cases of anorexia and bulimia were women. In addition to eating disorders, preoccupation with weight, body image and self-concept disturbances, are more prevalent among women than men.Women with eating disorders are also at risk for long-term psychological and social problems, including depression, anxiety, substance abuse and suicide. For instance, in 2000, the prevalence of depression among women who were hospitalized with a diagnosis of anorexia (11.5%) or bulimia (15.4 %) was more than twice the rate of depression (5.7 %) among the general population of Canadian women. The highest incidence of depression was found in women aged 25 to 39 years for both anorexia and bulimia. DATA GAPS AND RECOMMENDATIONS: Hospitalization data are the most recent and accessible information available. However, this data captures only the more severe cases. It does not include the individuals with eating disorders who may visit clinics or family doctors, or use hospital outpatient services or no services at all. Currently, there is no process for collecting this information systematically across Canada; consequently, the number of cases obtained from hospitalization data is underestimated. Other limitations noted during the literature review include the overuse of clinical samples, lack of longitudinal data, appropriate comparison groups, large samples, and ethnic group analysis. PMID- 15345085 TI - Violence against Canadian Women. AB - HEALTH ISSUE: Exposure to violence as children or as adults places a woman at higher risk of poor health outcomes, both physical and psychological. Abused women use more health care services and have poorer social functioning than non abused women. Knowledge of the prevalence of violence against women, and of which women are at risk, should assist in the planning of services for abuse prevention and treatment of the health consequences of abuse. KEY FINDINGS: The highest rates of any partner violence were in Alberta (25.5%) and British Columbia (23%). The lowest rates were in Ontario (18.8%). Women aged 15-24 had the highest rates in all regions in Canada, compared with older women. Aboriginal women in Manitoba/Saskatchewan and Alberta had higher rates of violence (57.2% and 56.6% respectively) than non-Aboriginal women (20.6%). Lower rates of partner-related violence were reported among women not born in Canada (18.4%) than among Canadian born women (21.7%). Visible minority women reported lower rates of lifetime sexual assault (5.7%) than non-visible minority women (12.3%). Perceptions of violence may vary by ethnicity. DATA GAPS AND RECOMMENDATIONS: More information is required concerning the prevalence of violence among Aboriginal women, immigrant and refugee women, women with disabilities, lesbian women and pregnant women. Future national population-based surveys need better questions on the health consequences of violence and related resource utilization. Further research is needed to identify the health care system's role in prevention, management and rehabilitation as they relate to violence against women. Future programs and policies must be based on valid, reliable and comprehensive empirical data. PMID- 15345086 TI - Perimenopausal and Postmenopausal Health. AB - HEALTH ISSUE: The average age of natural menopause in Western societies is estimated to be 51 years; women in Canada can therefore expect to live, on average, a third of their lives in post-menopausal years. During these years women are at increased risk of chronic diseases such as osteoporosis and cardiovascular disease. KEY FINDINGS: Clinical and epidemiological data on women in perimenopause are limited. There are no adequate Canadian data on symptom severity and prevalence among perimenopausal and postmenopausal women. Scientific evidence is lacking to support or refute claims that commonly used botanical products can offer therapeutic relief of menopausal symptoms.Recent data from the Women's Health Initiative suggest that combined estrogen plus therapy increases the risk of stroke, coronary artery disease and breast cancer. Hormone therapy is no longer recommended for the prevention of chronic diseases for asymptomatic women. Stroke is an important issue for perimenopausal and postmenopausal women and sex differences may exist in the progestin treatment of stroke. Osteoporosis affects an estimated one in six women over the age of 50. DATA GAPS AND RECOMMENDATIONS: There is a need to conduct clinical and epidemiological research aimed at better understanding the menopausal transition and defining its clinical phases. Investigations aimed at alternative combinations and doses of hormone therapy and non-pharmaceutical alternatives in light of known risks and benefits are also necessary. Health care practitioners and women need to be educated on the risks and effective treatment related to cardiovascular disease so they can present for treatment more quickly and receive the most effective therapies. PMID- 15345087 TI - Sexual Health. AB - HEALTH ISSUE: Much attention is devoted to women's reproductive health, but the formative and mature stages of women's sexual lives are often overlooked. We have analyzed cross-sectional data from the Sexual Behaviour module of the 2000/2001 Canadian Community Health Survey (CCHS), and reviewed the literature and available indicators of the sexual health of Canadian women. KEY FINDINGS: Contemporary Canadian adolescents are becoming sexually active at younger ages than in previous generations. The gender gap between young males and females in age at first intercourse has virtually disappeared. The mean age at first intercourse for CCHS respondents aged 15-24 years was between 16 and 17. Canadian born respondents are significantly younger at first intercourse than those who were born outside of Canada. Few adolescents recognize important risks to their sexual health. Older Canadians are sexually active, and continue to find emotional and physical satisfaction in their sexual relationships. DATA GAPS AND RECOMMENDATIONS: Both health surveys and targeted research must employ a broader understanding of sexuality to measure changes in and determinants of the sexual health of Canadians. There is reluctance to direct questions about sexual issues to younger Canadians, even though increased knowledge of sexual health topics is associated with delayed onset of sexual intercourse. Among adults, sex-positive resources are needed to address aspects of aging, rather than medicalizing age related sexual dysfunction. Age and gender-appropriate sexual health care, education, and knowledge are important not only for women of reproductive age, but for Canadians at all stages of life. PMID- 15345088 TI - Contraception. AB - HEALTH ISSUE: Contraception choices affect the long-term sexual health and fertility of women and men. Data from the 1998 Canadian Contraception Study and the 2000/2001 Canadian Community Health Survey were assessed for measures of contraceptive use and familiarity with various methods among Canadian women. KEY FINDINGS: The oral contraceptive (OC) pill is the dominant method of contraception for Canadian women. Canadian women demonstrate high awareness of the benefits of condom use, but 75% are unaware of the female condom. Among youth, condom use drops as OC use increases. Contraception use in sexually active females aged 15-17 is fairly high, but use is inconsistent. Sexually active adolescent females report high awareness of emergency contraception but poor knowledge of the time frame within which it is most effective. Women aged 35-44 are more familiar with and likely to choose sterilization than younger women. There has also been a shift away from tubal ligation in favour of vasectomies. DATA GAPS AND RECOMMENDATIONS: National data to guide policy and program development are limited. More data are needed on contraception use among males, and factors affecting accessibility, adherence and negotiation of choice. The importance of dual protection, and correct and consistent use of the chosen contraceptive method must be communicated to younger Canadians, as well as health care providers and educators. All women of reproductive age should be made aware of emergency contraception methods and increased efforts on sexual health promotion and education are required. Further research is essential to develop expanded contraceptive choices. PMID- 15345089 TI - Gender Differences in Bacterial STIs in Canada. AB - HEALTH ISSUE: The incidence of bacterial sexually transmitted infections (STIs) is rising in Canada. If these curable infections were prevented and treated, serious long-term sequelae including infertility, and associated treatment costs, could be dramatically reduced. STIs pose a greater risk to women than men in many ways, and further gender differences exist in screening and diagnosis. KEY FINDINGS: Reported incidence rates of chlamydia, gonorrhea, and infectious syphilis declined until 1997, when the trend began to reverse. The reported rate of chlamydia is much higher among women than men, whereas the reverse is true for gonorrhea and infectious syphilis. Increases in high-risk sexual behaviour among men who have sex with men were observed after the introduction of potent HIV suppressive therapy in 1996, but behavioural changes in women await further research. DATA GAPS AND RECOMMENDATIONS: STI surveillance in Canada needs improvement. Reported rates underestimate the true incidence. Gender-specific behavioural changes must be monitored to enhance responsiveness to groups at highest risk, and more research is needed on effective strategies to promote safer sexual practices. Geographic and ethnic disparities, gaps, and needs must be addressed. Urine screening for chlamydia should be more widely available for women as well as men, particularly among high-risk men in order to prevent re infections in their partners. As women are more likely to present for health examinations (e.g. Pap tests), these screening opportunities must be utilized. Female-controlled methods of STI prevention, such as safer topical microbicides, are urgently needed. PMID- 15345090 TI - Women and HIV. AB - HEALTH ISSUE: The epidemic of human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) in developed countries has changed from the early epidemic that affected primarily men who have sex with men, to one that increasingly affects other groups such as injecting drug users (IDU) and heterosexuals. As a result, the number and percentage of women with HIV and AIDS is increasing. KEY FINDINGS: The number of women in Canada living with HIV, including those with AIDS, has increased over time. An estimated 6,800 women were living with HIV at the end of 1999, an increase of 48.0 % from the 1996 estimate of 4,600. On an annual basis, women account for a growing proportion of positive HIV test reports among adults in Canada. This proportion increased from 10.7% in the period 1985-95 to 25% in 2001. Heterosexual contact is the main risk factor for HIV infection in women, accounting for 63% of newly diagnosed cases of HIV infection in adult Canadian women in 2001; the majority of the remainder is due to IDU. KEY DATA GAPS AND RECOMMENDATIONS: Research is needed to address specific information gaps regarding risk behaviours, testing patterns and HIV incidence and prevalence in women. This research needs to include the broader contextual factors that influence women's lives and their risk of HIV infection. Programmes and prevention efforts must be gender and age-specific and should target not only individual behaviours, but also the social and cultural context in which these behaviours occur. PMID- 15345091 TI - Perinatal Care in Canada. AB - HEALTH ISSUE: Canada's standard of perinatal care ranks among the highest in the world, but there is still room for improvement, both in terms of regional differences in care and global comparisons of approaches to care in Canada and elsewhere. Data from the Canadian Perinatal Surveillance System (CPSS) was used to evaluate morbidity and mortality among mothers and infants. KEY FINDINGS: Maternal mortality rates in Canada dropped to 4.4 per 100,000 live births in 1993 1997 and are among the lowest in the world. Rates of Caesarean section increased from 15.3 per 100 deliveries in 1994 to 19.1 in 1997. Although the infant mortality rate in Canada is among the lowest in the world (5.3-8.8 per 1,000 live births 1990-2000), there are unacceptable disparities between subpopulations. In Aboriginal populations, rates of stillbirth and perinatal mortality are 2-2.5 times the Canadian average. There has been a steady increase in the proportion of births among older women who have the highest risk of preterm births and pregnancy complications.The increasing rate of multiple births has accelerated recently and is of concern as these carry a higher risk of complications and are associated with an increased risk of preterm birth. The costs to the health care system are likely to be high. DATA GAPS AND RECOMMENDATIONS: CPSS data, including economic indicators, needs to be collected in a more timely and uniform manner across Canada. The CPSS should provide an evaluation of how well Canada fares in relation to international standards of perinatal care. PMID- 15345092 TI - Factors Associated with Women's Medication Use. AB - HEALTH ISSUE: Research has consistently shown that while women generally live longer than men, they report more illness and use of health care services (including medication). In the literature, the reasons for women's elevated medication use are not clear. This paper investigates the associations between over-the-counter (OTC) and prescription (Rx) medication use and selected social and demographic variables in men and women. KEY FINDINGS: While a larger proportion of women than men used medication throughout the study, the proportion of people using medication did not increase. The use of OTC and Rx medication increased by number of physician visits for women and men.Medication use increased with age, chronic disease and number of physician visits, and decreased with the perception of good to excellent health. The relationship with other factors varied for women and men depending on their education level, income and social roles. For women, the social roles of being married or previously married, being employed or being a parent did not increase their likelihood of medication use. Reported income adequacy is not associated with the chances of mediation use among highly educated women, but for women with low levels, medication use increases as income adequacy decreases. DATA GAPS AND RECOMMENDATIONS: More complete data are needed about social roles and their relation to mediation use. Data that would allow an assessment of the appropriateness of OTC and Rx drug use or the reasons for such use need to be collected. More research is needed to better understand the distribution and determinants of specific medication use. PMID- 15345093 TI - Synthesis : Pulling It All Together. PMID- 15345094 TI - Women's Health Surveillance: Implications for Policy. PMID- 15345095 TI - Integrating Ethnicity and Migration As Determinants of Canadian Women's Health. AB - HEALTH ISSUE: This chapter investigates (1) the association between ethnicity and migration, as measured by length of residence in Canada, and two specific self reported outcomes: (a) self-perceived health and (b) self-reports of chronic conditions; and (2) the extent to which these selected determinants provide an adequate portrait of the differential outcomes on Canadian women's self-perceived health and self-reports of chronic conditions. The 2000 Canadian Community Health Survey was used to assess these associations while controlling for selected determinants such as age, sex, family structure, highest level of education attained and household income. KEY FINDINGS: * Recent immigrant women (2 years or less in Canada) are more likely to report poor health than Canadian-born women (OR = 0.48 CI: 0.30-0.77). Immigrant women who have been in Canada 10 years and over are more likely to report poor health than Canadian-born women (OR = 1.31 CI: 1.18-1.45).* Although immigrant women are less likely to report chronic conditions than Canadian-born women, this health advantage decreased over time in Canada (OR from 0.35 to 0.87 for 0-2 years to 10 years and above compared with Canadian born women). DATA GAPS AND RECOMMENDATIONS: * Migration experience needs to be conceptualized according to the results of past studies and included as a social determinant of health above and beyond ethnicity and culture. It is expected that the upcoming longitudinal survey of immigrants will help enhance surveillance capacity in this area.* Variables need to be constructed to allow women and men to best identify themselves appropriately according to ethnic identity and number of years in the host country; some of the proposed categories used as a cultural group may simply refer to skin colour without capturing associated elements of culture, ethnicity and life experiences. PMID- 15345096 TI - Health Care Utilization by Canadian Women. AB - HEALTH ISSUES: While women are reported to be more frequent users of health services in Canada, differences in women's and men's health care utilization have not been fully explored. To provide an overview on women's healthcare utilization, we selected two key issues that are important for public policy purposes: access to care and patterns of utilization. These issues are examined using primarily data from the 1998/99 National Population Health Survey, complemented by the 2000 Canadian Community Health Survey and the 2001 Health Service Access Survey. KEY FINDINGS: * Women are twice as likely as men to report a regular family physician, but that proportion is very low (15.8%).* Women report significantly shorter specialist wait times (20.9 days) than men (55.4 days) for mental health, while the reverse is true for asthma and other breathing conditions (10.8 for men, 78.8 for women).* Reported mean wait times are significantly lower for men than for women pertaining to overall diagnostic tests: for MRI, 70.3 days for women compared to 29.1 days for men. DATA GAPS AND RECOMMENDATIONS: * Measurement of possible system bias and its implication for equitable and quality healthcare for women requires larger provincial samples of the national surveys, along with a longitudinal design.* Either a national database on preventive services, or better alignment of provincial databases pertaining to health promotion and preventive services, is needed to facilitate data linkage with national surveys to undertake longitudinal studies that support gender based analyses.en are reported to be more frequent users of health services in Canada, differences in women's and men's health care utilization have not been fully explored. To provide an overview on women's healthcare utilization, we selected two key issues that are important for public policy purposes: access to care and patterns of utilization. These issues are examined using primarily data from the 1998/99 National Population Health Survey, complemented by the 2000 Canadian Community Health Survey and the 2001 Health Service Access Survey. KEY FINDINGS: * Women are twice as likely as men to report a regular family physician, but that proportion is very low (15.8%).* Women report significantly shorter specialist wait times (20.9 days) than men (55.4 days) for mental health, while the reverse is true for asthma and other breathing conditions (10.8 for men, 78.8 for women).* Reported mean wait times are significantly lower for men than for women pertaining to overall diagnostic tests: for MRI, 70.3 days for women compared to 29.1 days for men. DATA GAPS AND RECOMMENDATIONS: * Measurement of possible system bias and its implication for equitable and quality healthcare for women requires larger provincial samples of the national surveys, along with a longitudinal design.* Either a national database on preventive services, or better alignment of provincial databases pertaining to health promotion and preventive services, is needed to facilitate data linkage with national surveys to undertake longitudinal studies that support gender based analyses. PMID- 15345097 TI - Integrating Socio-Economic Determinants of Canadian Women's Health. AB - HEALTH ISSUE: The association between a number of socio-economic determinants and health has been amply demonstrated in Canada and elsewhere. Over the past decades, women's increased labour force participation and changing family structure, among other changes in the socio-economic environment, have altered social roles considerably and lead one to expect that the pattern of disparities in health among women and men will also have changed. Using data from the CCHS (2000), this chapter investigates the association between selected socio-economic determinants of health and two specific self-reported outcomes among women and men: (a) self-perceived health and (b) self-reports of chronic conditions. KEY FINDINGS: The descriptive picture demonstrated by this CCHS dataset is that 10% of men aged 65 and over report low income, versus 23% of women within the same age bracket. The results of the logistic regression models calculated for women and men on two outcome variables suggest that the selected socio-economic determinants used in this analysis are important for women and for men in a differential manner. These results while supporting other results illustrate the need to refine social and economic characteristics used in surveys such as the CCHS so that they would become more accurate predictors of health status given that there are personal, cultural and environmental dimensions to take into account. RECOMMENDATIONS: Because it was shown that socio economic determinants of health are context sensitive and evolve over time, studies should be designed to examine the complex temporal interactions between a variety of social and biological determinants of health from a life course perspective. Examples are provided in the chapter. PMID- 15345098 TI - Is acupuncture a useful adjunct to physiotherapy for older adults with knee pain?: the "acupuncture, physiotherapy and exercise" (APEX) study [ISRCTN88597683]. AB - BACKGROUND: Acupuncture is a popular non-pharmacological modality for treating musculoskeletal pain. Physiotherapists are one of the largest groups of acupuncture providers within the NHS, and they commonly use it alongside advice and exercise. Conclusive evidence of acupuncture's clinical effectiveness and its superiority over sham interventions is lacking. The Arthritis Research Campaign (arc) has funded this randomised sham-controlled trial which addresses three important questions. Firstly, we will determine the additional benefit of true acupuncture when used by physiotherapists alongside advice and exercise for older people presenting to primary care with knee pain. Secondly, we will evaluate sham acupuncture in the same way. Thirdly, we will investigate the treatment preferences and expectations of both the participants and physiotherapists participating in the study, and explore the effect of these on clinical outcome. We will thus investigate whether acupuncture is a useful adjunct to advice and exercise for treating knee pain and gain insight into whether this effect is due to specific needling properties. METHODS/DESIGN: This randomised clinical trial will recruit 350 participants with knee pain to three intervention arms. It is based in 43 community physiotherapy departments in 21 NHS Trusts in the West Midlands and Cheshire regions in England. Patients aged 50 years and over with knee pain will be recruited. Outcome data will be collected by self-complete questionnaires before randomisation, and 6 weeks, 6 months and 12 months after randomisation and by telephone interview 2 weeks after treatment commences. The questionnaires collect demographic details as well as information on knee-related pain, movement and function, pain intensity and affect, main functional problem, illness perceptions, self-efficacy, treatment preference and expectations, general health and quality of life. Participants are randomised to receive a package of advice and exercise; or this package plus real acupuncture; or this package plus sham acupuncture. Treatment details are being collected on a standard proforma. Interventions are delivered by experienced physiotherapists who have all received training in acupuncture to recognised national standards. The primary analysis will investigate the main treatment effects of real or sham acupuncture as an adjunct to advice and exercise. DISCUSSION: This paper presents detail on the rationale, design, methods, and operational aspects of the trial. PMID- 15345099 TI - Lactobacillus casei strain GG in the treatment of infants with acute watery diarrhea: a randomized, double-blind, placebo controlled clinical trial [ISRCTN67363048]. AB - BACKGROUND: Adjuvant therapy to ORT with probiotic bacteria for infants with acute watery diarrhea has been under active investigation. Most studies have been done in the developed world showing benefit only for viral mild gastroenteritis. We evaluated the effect of a milk formula containing one billion (109) cfu/ml of Lactobacillus casei strain GG (LGG) upon duration and severity of diarrhea in infants in an environment with more severe acute diarrhea, where etiologic agents other than rotavirus are involved more frequently, and where mixed infections are more prevalent. METHODS: Male infants aged 3-36 months brought for treatment of acute watery diarrhea of less than 48 hours were eligible. After rehydration was completed with the WHO's oral rehydration solution, patients were randomly assigned to receive a milk formula either containing LGG or not. Stool volume was periodically measured using a devise suited to collect stools separate from urine. Duration of diarrhea was estimated based on stools physical characteristics. RESULTS: Eighty nine patients received the placebo milk formula and ninety received the LGG containing formula. Both groups were comparable in their baseline characteristics. Total stool output was significantly larger (p = 0.047) in the LGG group (247.8 ml/kg) than in the placebo group (195.0 ml/kg). No significant differences were found in duration of diarrhea (58.5 hours with LGG vs. 50.4 hours with placebo), rate of treatment failure (21.1% with LGG vs. 18.0% with placebo), and proportion of patients with unresolved diarrhea after 120 hours (12.2% with LGG vs. 12.5% with placebo). The rate of stools with reducing substances after 24 hours of treatment increased significantly in both groups (from 41.4% to 72.2% with LGG and from 45.9% to 68.0% with placebo). CONCLUSION: This study did not show a positive effect of LGG on the clinical course of acute watery diarrhea. Positive beneficial effects of LGG, as had been reported elsewhere, could have been masked in our study by worsening diarrhea due to transient lactose malabsorption. Further studies with low-lactose or non-lactose conveyors of LGG are desirable. PMID- 15345100 TI - Certificates of confidentiality. PMID- 15345101 TI - Impact of genetic counseling and testing on colorectal cancer screening behavior. PMID- 15345102 TI - Preconceptional cystic fibrosis carrier screening: attitudes and intentions of the target population. AB - The aim of this study was to assess the attitudes and intentions of individuals planning a pregnancy with regard to preconceptional cystic fibrosis (CF) carrier screening and to determine factors associated with a positive and negative/neutral intention to have the test. A survey, based on a questionnaire, was conducted among a stratified random sample of 303 recently married couples (606 individuals). Of the eligible individuals, 70% (n = 380) participated. Of the respondents, 73% had a positive attitude toward a routine offer of preconceptional CF carrier screening, and 56% had the intention to participate in a screening program. A positive intention to have the test was associated with high perceived anticipation of regret, intended preconceptional behavior, high perceived pressure from experts, high perceived consequences of the test results, low perceived barriers, and low perceived negative consequences for family members. These results suggest that the offer of routine preconceptional CF carrier screening would lead to substantial acceptance among couples planning a pregnancy. Several variables related with intention were identified. PMID- 15345103 TI - Attitudes about and psychosocial outcomes of HFE genotyping for hemochromatosis. AB - We examined attitudes regarding genetic testing and psychosocial outcomes of HFE genotyping for hemochromatosis. A total of 87 persons with hemochromatosis (patients) (39 women, 48 men), who underwent HFE genotyping, and 50 persons with hypertension (controls) (22 women, 28 men), who had not undergone HFE genotyping, completed a structured interview in which they reported attitudes about benefits and disadvantages of genetic testing and their understanding of genetics and hemochromatosis. Among patients, adherence to treatment for hemochromatosis was assessed. Controls estimated the likelihood of experiencing several potential positive and negative psychosocial outcomes after a positive genetic test. Patients reported their experience pertinent to these outcomes. Patients received information about hemochromatosis when they were diagnosed, and controls read a brief description of hemochromatosis before answering questions. Patients correctly answered 65% of knowledge questions and controls correctly answered 59%. Most participants believed genetic testing is beneficial and described few negative aspects of testing. Controls expected to experience more anxiety, depression, and anger related to a positive genetic test than was reported by patients (p < 0.001). One patient reported discrimination related to the HFE genotype. Most patients were compliant with the iron depletion and maintenance phases of treatment for hemochromatosis. Race, sex, marital status, income, education, barriers to treatment, and knowledge were not significantly associated with adherence to maintenance phlebotomy. We conclude that HFE genotyping appears to be viewed positively and would be generally accepted were it offered as part of a screening program for hemochromatosis. Persons who have not undergone genetic testing may overestimate their emotional responses to a positive test result. In the present hemochromatosis patients, few reported that HFE genotyping was accompanied by negative psychosocial outcomes. PMID- 15345104 TI - Hereditary hemochromatosis genetic testing of at-risk children: what is the appropriate age? AB - The objective of this study was to consider the objective evidence and ethical arguments for the appropriate age to test children at risk of developing hereditary hemochromatosis. A literature search for information on iron overload in children, onset of disease expression for hemochromatosis, and recommendations for age of cascade screening was undertaken. We examined the objective evidence and arguments for testing in early childhood and those for delaying testing until later teenage years. Cascade testing of offspring of people with hemochromatosis is widely advocated because it is an easily preventable disease. The ideal age to test those offspring is a matter of debate. Some authorities advocate testing at a very young age whereas others recommend delaying testing until late teenage years. To date there has been no published overview of the objective evidence and arguments central to this debate. In children who are C282Y homozygous, iron overload is rare in the first two decades of life and associated morbidity has only been documented in 1 patient. In the cascade setting, genetic testing for hemochromatosis need not be offered until late teenage years. PMID- 15345105 TI - Rapid detection of common southeast Asian beta-thalassemia mutations by nonisotopic multiplex PCR-SSCP analysis. AB - This report describes the detection of seven beta-thalassemia mutations common in Southeast Asia by amplifying three short PCR fragments in two separate tubes, followed by single-strand conformation polymorphism (SSCP) analysis in single lanes. These mutations are -28 A --> G, codon 17 A --> T, IVS1 + 5 G --> C, codon 41/42 -CTTT, codon 43 G --> T, codon 71/72 + A, and IVS2 + 654 C --> T, and account for 70% to over 95% of the cases in this region. This rapid nonisotopic method was also found capable of detecting other mutations within the amplified fragments. It is simple, rapid, and cheap, and thus suitable for carrier screening and prenatal diagnosis in Southeast Asia. PMID- 15345106 TI - Informing carriers of beta-thalassemia: giving the good news. AB - This study explored the value of informing beta-thalassaemia carriers of the advantages, as well as the disadvantages of carrier status. Twenty-eight carriers of beta-thalassaemia were interviewed immediately after counselling, and again 2 weeks later. Both interviews included administration of a psychological scale (previously used for cystic fibrosis). Immediately after the first interview the intervention group (n = 18) were informed of the protective effect of the beta thalassaemia trait against malaria and coronary heart disease. The control group (n = 10) was given the same information after the second interview. The effect of giving the positive information was assessed by comparing participants' scores at the first and second interview. Knowledge of carrier status aroused several negative feelings, including shock, sadness, and anger, but little feeling of stigmatization. Two weeks later, negative feelings were unchanged in the control group, but they were reduced in all members of the intervention group. All members of the intervention group considered it important to inform carriers of the positive aspects as well as the risks associated with carrier status. Carriers of recessive disorders with a known heterozygote advantage should be informed of the advantage. This information has now been incorporated into the comprehensive information system for hemoglobin disorders available at http://www.chime.ucl.ac.uk/ApoGI/. PMID- 15345107 TI - Prevalence of major depression one year after predictive testing for Huntington's disease. AB - Psychiatric hospitalizations, completed suicides, and suicide attempts are rare after predictive testing for Huntington's disease (HD). Case studies have shown that major depression can be a consequence of being tested, although no studies have shown how common this is. The present study evaluated the prevalence of major depression during the first year after disclosure. We conducted retrospective data and chart reviews of 153 persons (50 testing positive, 103 testing negative) evaluated every 3 months for depression. There was no significant baseline difference in the percentage of "positives" and "negatives" who had pre-testing major depressive episodes (14% vs. 12%, respectively). A senior psychiatrist reviewed data from the Schedule for Affective Disorders and Schizophrenia-Change Version, from the Beck Depression Inventory, and from clinical notes for every follow-up contact completed. The 1-year prevalence of major depression among positives was 6.0%, compared to 3.0% among negatives (p = 0.30), and an estimated 3% population prevalence. One-year prevalence of clinically significant depressive symptoms, whether or not major depression was diagnosed, was 20.0% in positives and 12.6% in negatives (p = 0.17). Although not statistically significant, depressive symptoms and major depression occurred more frequently among those who tested positive. Despite some evidence to the contrary, including our own studies, a positive predictive test for HD is not psychologically benign. Clinical testing programs should assess patients for depressive symptoms after testing, and patients with clinically significant complaints should be referred to a mental health professional. PMID- 15345108 TI - The role of test accuracy in predicting acceptance of genetic susceptibility testing for Alzheimer's disease. AB - A survey questionnaire regarding perceptions of risk and genetic susceptibility to Alzheimer's disease (AD) was completed by 518 offspring of AD cases from families with multiple affected, ascertained as part of a genetic linkage study of late onset AD. The questionnaire focused on respondents' perceptions of their own risk for AD as well as on the properties of real and hypothetical susceptibility tests, including error rates for false-positive and false-negative test results. Our findings showed that about 20% of the sample would refuse a susceptibility test with zero error rates, about 40% would accept tests with very high error rates in both directions, and the remainder would exercise some discrimination. Acceptance of high test error rates was significantly associated with male gender, low education, and high perceived lifetime risk of AD. In a previous paper related to this work, we showed that physicians caring for these families exercised much more discrimination in judging the acceptability of genetic tests they would offer to these same respondents. The findings show that there is a pressing need to educate the public, particularly those with relatives affected by a complex disease, to expect standards of accuracy for genetic tests comparable to those that prevail in other diagnostic and prognostic testing efforts in the broad field of clinical medicine. PMID- 15345109 TI - BRCA1 mutations in a population-based study of breast cancer in Stockholm County. AB - The mutation frequency of BRCA1 and BRCA2 in women with breast cancer varies according to family history, age at diagnosis and ethnicity. The contribution of BRCA1 and BRCA2 mutations in breast cancer populations, unselected for age and family history, has been examined in several studies reporting mutation frequencies between 1% and 12% by screening methods, population sizes, and to what extent the gene/s were screened differed in the studies. We wanted to clarify the proportion of breast cancer attributable to mutations in BRCA1 in an unselected breast cancer population from the Stockholm region. All incident breast cancer patients treated surgically in a 19-month period were eligible for the study and 70% (489/696) participated. Exon 11 of BRCA1 was screened for mutations using the protein truncation test, and the mutation frequency was estimated from that. In previous studies on high-risk families from Stockholm, more than 70% of the mutations were detected in exon 11. Two mutations were found, both in patients with a family history or their own medical history of ovarian cancer, giving a mutation frequency in exon 11 of 0.4% and an estimated BRCA1 mutation frequency of <1%. Mutations in BRCA1 in unselected breast cancer cases in our region are rare and likely to be found only in high-risk families. Our BRCA1 prevalence is the lowest of all studies on unselected breast cancer patients, probably reflecting the comparatively low rates detected also in high risk breast cancer families from the region. PMID- 15345110 TI - Characterization of two novel BRCA1 germ-line mutations involving splice donor sites. AB - Deleterious BRCA1 mutations have significant clinical implications for the patients that carry them. Point mutations in critical functional domains and frameshift mutations that lead to early termination of protein translation are associated with a 60-80% risk of breast cancer and a 20-40% risk of ovarian cancer. In contrast, the significance of mutations located in intronic regions of BRCA1, even in the setting of a family history of breast and ovarian cancer, is not always clear. Some of these mutations occur in splice donor/acceptor consensus sites. These mutations can affect heteronuclear RNA (hnRNA) processing, leading to the loss of functional BRCA1 protein and thus may be disease associated. However, it is important to verify the effect of these mutations, because splicing alterations cannot be predicted from genomic sequence alone. We report here the characterization of two novel BRCA1 mutations identified in families seen in our cancer risk evaluation clinic that alter splice donor sites of BRCA1. We show that both mutations alter transcript splicing and result in truncated BRCA1. IVS17 + 1G --> T leads to inclusion of part of intron 17 after the coding sequence of exon 17, resulting in early termination of BRCA1 protein following codon 1692. 252del5insT abolishes the splice donor site in exon 3, leading to the skipping of exon 5 and BRCA1 protein truncation following codon 45. Thus, both mutations result in loss of BRCA1 function, and carriers of these mutations should be counseled in the same manner as carriers of other truncating BRCA1 mutations. PMID- 15345111 TI - Subjective and objective risk of breast cancer in Ashkenazi Jewish individuals at risk for BRCA1/2 mutations. AB - The aims of the study were to (1) examine the differences between subjective and objective estimates of the risk of breast cancer in those being tested for BRCA1/2 mutations, (2) explore new ways to conceptualize risk, and (3) examine the change in subjective risk of developing breast cancer throughout the process of genetic counseling and testing. Participants were 86 Ashkenazi Jewish women with a family or personal history indicating risk for BRCA1/2 mutations. Surveys to assess subjective risk of breast cancer (percentage risk, projected age of onset, and survival time) were administered before counseling, after counseling, and after receipt of test results. Subjective percentage risk of breast cancer was compared to estimated objective risk to determine accuracy. Those with no personal history of cancer receiving positive results became more accurate from post-counseling to post-result. Those receiving positive results increased their estimate of their percentage risk, and those receiving uninformative negative results decreased their estimate of their percentage risk from post-counseling to post-result. Those without a personal history of cancer decreased in perceived risk from post-counseling to post-result. No change in projected age of onset of breast cancer or survival time with breast cancer was seen from pre- to post counseling or from post-counseling to post-result, and no change in accuracy or in percentage risk of breast cancer was seen from pre- to post-counseling. Individuals use information from genetic counseling to form estimates of percentage risk following receipt of test results; however, projected age of onset and survival time with breast cancer, areas not targeted by genetic counseling that may be more closely linked to health behavior, do not change. PMID- 15345112 TI - High-risk premenopausal women's experiences of undergoing prophylactic oophorectomy: a descriptive study. AB - Women who are at increased risk of developing ovarian cancer because of their family history may need to make decisions about the medical management of their cancer risk--whether to have ovarian screening or undergo prophylactic surgery. This qualitative study explores the perceived physical and emotional implications of undergoing preventative surgery using data collected during interviews with 23 high-risk premenopausal women who had undergone prophylactic oophorectomy because of their family history of cancer. Despite the fact that all of these women regarded their decision to undergo surgery extremely positively, 20 women also described what they regarded as the costs of undergoing surgery. These included post-operative complications, the onset of menopausal symptoms, side effects of hormone replacement therapy, and negative effects on body image and gender identity. The perceived benefits of surgery were described as risk reduction, enabling one to fulfil family obligations, removing the need for gynecological screening, cessation of menstruation, and positive side effects of hormone replacement therapy. This study suggests there is a need to inform women about potential physical and emotional sequelae of oophorectomy prior to undergoing this procedure. PMID- 15345113 TI - Mutation analysis of hMSH2 and hMLH1 in colorectal cancer patients in India. AB - The aim of this work was to study the mutation profile in hMSH2 and hMLH1 genes in hereditary nonpolyposis colorectal cancer (HNPCC) patients in India. On the basis of the Bethesda criteria, 31 colorectal cancer patients were studied first for microsatellite instability, using the five markers recommended by the Bethesda guidelines. Twelve of 31 tumor samples were found to be MSI-H, 9 of 31 were MSI-L, and the rest were MSS. The 12 patients with MSI-H were analyzed for mutations in hMSH2 and hMLH1 genes using PCR-denaturing high-performance liquid chromatography (dHPLC), followed by sequencing of samples showing abnormal peaks. Of the five mutations detected, three were found to be deleterious mutations (hMSH2-R680X, hMLH1-E671X, and a splice junction mutation IVS16-2A --> G); one had a mutation of probable significance (hMLH1-C680G) and one was of unknown significance (hMSH2-R171K). This study has also shown that most of the early onset colon (4/7) and early-onset rectal (15/21) cancers are MSS or MSI-L. This is the first study to describe the mutation in hMSH2 and hMLH1 in Indian patients, a low incidence region for colorectal cancer. A two-stage procedure using MSI testing followed by PCR-dHPLC was found to be an efficient method in studying the mutation profile in high-risk patients. PMID- 15345114 TI - Description of the first two seemingly unrelated Greek Cypriot families with a common C618R RET proto-oncogene mutation. AB - Germ-line mutations of the RET proto-oncogene cause three different cancer syndromes: multiple endocrine neoplasia type 2A (MEN2A), multiple endocrine neoplasia type 2B, and familial medullary thyroid carcinoma (FMTC). The objective of the present study was the clinical and molecular characterization of the first two Greek Cypriot families diagnosed with MEN2A and FMTC. The clinical diagnosis of the probands was based on clinical presentation and supported with laboratory findings (calcitonin and carcinoembryonic antigen tumor marker levels). We screened the RET gene by direct DNA sequencing of exons 10, 11, and 16 using genomic DNA as templates. After identification of the mutation, we also developed the amplification refractory mutation system (ARMS) as an alternative method to direct sequencing for genetic diagnosis of 22 additional individuals from both families. We identified the germ-line missense mutation T --> C of codon 618 of exon 10 (C618R) in the probands of both families. By using ARMS, two members of the MEN2A family and five members of the FMTC family were also found positive for the C618R mutation. These are the first seemingly unrelated families in Cyprus investigated clinically and molecularly in detail and shown to transmit this common RET proto-oncogene mutation. PMID- 15345115 TI - Attitude toward genetic testing for cancer risk in Istanbul. AB - To identify attitudes toward genetic testing, and the effects of this information on decisions regarding issues such as pregnancy, abortion, and prophylactic surgery, several subsets of the Turkish population were surveyed in hospital settings. Individuals (n = 179) chosen arbitrarily from four different subsets of a Turkish population were asked to participate in a confidential 23-question survey. Survey participants were familiar with the concept of cancer being a familial disease (85.5%), and 84.7% of them expressed interest in genetic testing to determine cancer risk, 83.9% would have their fetuses tested for such cancer risk, 65.1% would terminate their pregnancies, 92.2% would have their children tested if they were determined to have an increased cancer risk, 71.9% would agree to undergo prophylactic oophorectomy or orchiectomy and 67.6% would have mastectomy/prostatectomy should there be an increased cancer risk to these organs. It appears that at least the sampled segment of a Turkish population is willing to undergo genetic testing to determine if they are at increased risk for cancer. The feasibility and acceptance of genetic testing and the influence of education and genetic counseling in the Turkish people should further be evaluated with a larger stratified sample of the population. PMID- 15345116 TI - Heterozygosity for Tay-Sachs and Sandhoff diseases in non-Jewish Americans with ancestry from Ireland, Great Britain, or Italy. AB - Previous reports have found that non-Jewish Americans with ancestry from Ireland have an increased frequency of heterozygosity for Tay-Sachs disease (TSD), although frequency estimates are substantially different. Our goal in this study was to determine the frequency of heterozygosity for TSD and Sandhoff diseases (SD) among Irish Americans, as well as in persons of English, Scottish, and/or Welsh ancestry and in individuals with Italian heritage, who were referred for determination of their heterozygosity status and who had no known family history of TSD or SD or of heterozygosity for these conditions. Of 610 nonpregnant subjects with Irish background, 24 TSD heterozygotes were identified by biochemical testing, corresponding to a heterozygote frequency of 1 in 25 (4%; 95% CI, 1/39-1/17). In comparison, of 322 nonpregnant individuals with ancestry from England, Scotland, or Wales, two TSD heterozygotes were identified (1 in 161 or 0.62%; 95% CI, 1/328-1/45), and three TSD heterozygotes were ascertained from 436 nonpregnant individuals with Italian heritage (1 in 145 or 0.69%; 95% CI, 1/714-1/50). Samples from 21 Irish heterozygotes were analyzed for HEXA gene mutations. Two (9.5%) Irish heterozygotes had the lethal + 1 IVS-9 G --> A mutation, whereas 9 (42.8%) had a benign pseudodeficiency mutation. No mutation was found in 10 (47.6%) heterozygotes. These data allow for a frequency estimate of deleterious alleles for TSD among Irish Americans of 1 in 305 (95% CI, 1/2517 1/85) to 1 in 41 (95% CI, 1/72-1/35), depending on whether one, respectively, excludes or includes enzyme-defined heterozygotes lacking a defined deleterious mutation. Pseudodeficiency mutations were identified in both of the heterozygotes with ancestry from other countries in the British Isles, suggesting that individuals with ancestry from these countries do not have an increased rate of TSD heterozygosity. Four SD heterozygotes were found among individuals of Italian descent, a frequency of 1 in 109 (0.92%; 95% CI, 1/400-1/43). This frequency was higher than those for other populations, including those with Irish (1 in 305 or 0.33%; 95% CI, 1/252-1/85), English, Scottish, or Welsh (1 in 161 or 0.62%; 95% CI, 1/1328-1/45), or Ashkenazi Jewish (1 in 281 or 0.36%; 95% CI, 1/1361-1/96) ancestry. Individuals of Irish or Italian heritage might benefit from genetic counseling for TSD and SD, respectively. PMID- 15345117 TI - GJB2 and GJB6 mutations in 165 Danish patients showing non-syndromic hearing impairment. AB - Thirty-two genes causing non-syndromic hearing impairment (NSHI) have been cloned, including GJB2 and GJB6 encoding the gap junction subunits connexin 26 and connexin 30, respectively. One mutation in GJB2, 35delG, accounts for a large percentage of GJB2 hearing impairment in Southern Europe whereas a considerably lower frequency has been reported from Northern European populations. Recently, a 342-kb deletion implicating GJB6 was found in 22 out of 44 NSHI patients of Spanish origin with only one mutated allele of GJB2. We report the first study of GJB2 and GJB6 mutations in Danish patients with NSHI. We tested 165 individuals and found GJB2 mutations in 16 individuals. The deletion implicating GJB6 was found in two individuals out of 9 heterozygous for GJB2 mutation. Furthermore, we screened 509 unselected samples from the Danish newborn population for the 35delG mutation in GJB2. We found 9 samples heterozygous for 35delG and 11 samples heterozygous for mutations leading to amino acid variants in GJB2 protein. In conclusion, our data are in accordance with results from other Northern European populations. Furthermore, our data on the GJB6 deletion suggest that routine screening for this deletion could help to explain hearing impairment in some Northern European NSHI patients heterozygous for a mutation in GJB2. PMID- 15345118 TI - Rapid combined genotyping assay for four achondroplasia and hypochondroplasia mutations by real-time PCR with multiple detection probes. AB - Achondroplasia (ACH) and hypochondroplasia (HYCH) are the most prevalent genetic short-stature syndromes. Whereas the diagnosis of ACH can be established on clinical and radiologic grounds alone in the majority of cases, HYCH is more difficult to confirm. Molecular genetic analysis of both skeletal dysplasias can be especially helpful for the purpose of prenatal diagnosis, in early childhood to differentiate definitively between the largely overlapping phenotypes, and in atypical presentations. The two most prevalent mutations for each syndrome cause substitution of a single respective nucleotide. These mutations can be identified by a variety of molecular methods, including PCR with restriction enzyme digestion or direct DNA sequencing. We have developed a single-step, real-time PCR assay in which two detection probes are applied in combination with a single anchor probe at each mutation position. Because the two most prevalent mutations for each syndrome cause substitution of a single respective nucleotide, this approach guarantees optimal differentiation during probe dissociation analysis after amplification. This assay, which is performed on the LightCycler thermocycler, enables the rapid and reliable detection of the two most common FGFR3 mutations associated with ACH (1138G --> A and 1138G --> C; G380R) and HYCH (1620C --> A and 1620 C --> G; N540K) in a single test. PMID- 15345119 TI - PCR-RFLP genotyping assay for a lactase persistence polymorphism upstream of the lactase-phlorizin hydrolase gene. AB - The majority of the world's human population experiences a decline of lactase gene expression during maturation, so-called lactase nonpersistence. Thus, adults with lactase nonpersistence are susceptible to developing symptoms of lactose intolerance. By contrast, lactase persistence is an autosomal dominant heritable condition that results in a high level of lactase gene expression throughout adulthood and sustained lactose tolerance. Lactase persistence has recently been correlated with a single nucleotide genetic variant (a C --> T mutation) located 13,910 bases upstream from the lactase structural gene. We aimed to develop a restriction fragment length polymorphism (RFLP) method of detecting the C/T variants as a means of identifying individuals genetically inclined toward lactase persistence or nonpersistence. Genomic DNA in a 210-bp region surrounding the -13,910-bp variant site was PCR amplified with unique primers designed to avoid or mutate adjacent restriction sites. The amplified DNA was digested with a restriction enzyme, CviJI, that recognizes the base pair sequence generated by the lactase nonpersistence variant. Restriction digest gel analysis yielded DNA fragments of the expected diagnostic molecular weight sizes for individuals that were homozygote or heterozygote for the lactase persistence and nonpersistence variants. The genotypes predicted by the RFLP-based method were confirmed by DNA sequence analysis. The RFLP-based method provides a quick and noninvasive means of molecular detection of the presence or absence of the lactase persistence variant. PMID- 15345120 TI - Comprehensive analysis of genetic polymorphisms in the interleukin-10 promoter: implications for immune regulation in specific ethnic populations. AB - The association of interleukin-10 (IL-10) promoter single-nucleotide polymorphisms (SNPs) as risk factors for certain inflammatory diseases, viral infections, cancers, and transplant rejection have been the subject of recent studies. The SNPs -1082 G --> A, -819 C --> T, and -592 C --> A, which have been associated with differential IL-10 production, are strongly linked with ethnicity. In this study, we determined the ethnic distribution of IL-10 promoter SNPs and their haplotype rates among Hispanics, African Americans, and Caucasians from Texas and Ashkenazi Jews from New York. Significant differences in prevalence rates of IL-10 SNPs (and their haplotype distribution) were found. African Americans and Hispanics have a lower rate of putative high-producer SNPs and a higher rate of low IL-10 producers when compared to Caucasians or Ashkenazi Jews. No statistically significant differences in allelic frequencies and haplotype rates were observed between Caucasians and Ashkenazi Jews. This study provides critical new information on the ethnic distribution of IL-10 promoter SNPs in a regional U. S. population and is the first to analyze the rate of SNPs in an unstudied ethnic population, Ashkenazi Jews. Knowledge of IL-10 promoter polymorphisms may prove useful in prediction of immunization responses, disease severity, and in the intelligent design of customized immunotherapy. PMID- 15345121 TI - Patterns of chromosomal translocations identified by a birth defects registry, Hawaii, 1986-2000. AB - Using a birth defects registry, this investigation examined the distribution of translocations by type of translocation, chromosomes involved in the translocation, pregnancy outcome, method of diagnosis, inheritance, and diagnosis of major structural birth defects. A total of 121 cases were identified through a statewide population-based birth defects registry. The translocations were reciprocal in 89 (73.6%) cases, Robertsonian in 32 (26.4%) cases, balanced in 86 (71.1%) cases, and unbalanced in 35 (28.9%) cases. Live births accounted for 76 (88.4%) of balanced translocations and 22 (62.9%) of unbalanced translocations. Diagnosis was made by amniocentesis or chorionic villus sampling in 72 (83.7%) of balanced translocations and 11 (31.4%) of unbalanced translocations. Of cases of known inheritance, the translocation was of maternal origin in 38 (46.3%) cases, paternal origin in 25 (30.5%) cases, and de novo in 19 (23.2%) cases. Major structural birth defects were diagnosed in 17 (19.8%) of balanced translocations and 20 (57.1%) of unbalanced translocations. Translocations were more likely to be reciprocal, balanced, and of maternal origin. Infants and fetuses with unbalanced translocations were less likely to be live births and diagnosed by amniocentesis or chorionic villus sampling and more likely to be diagnosed with major structural birth defects. PMID- 15345122 TI - Human subject protections in genetic research. AB - The Certificate of Confidentiality (COC) is a voluntary tool to protect researchers from being compelled to release identifying information about their subjects. Institutional review board (IRB) review and informed consent (IC) procedures are mandatory tools to protect human subjects. Although many studies reveal poor documentation of IRB and IC procurement, most published research undergoes IRB review and has appropriate IC procedures. There are no empirical data about the use of COCs. We examined the procurement and documentation of all these human subject protections in the genetics literature. A total of 112 (55%) articles documented IRB review, 108 (53%) document IC, and 82 (41%) documented both. None documented the procurement of a COC. Returned surveys provided additional information that confirmed that at least 74% (n = 150) of research had received appropriate IRB review, at least 71% (n = 143) had procured IC, and at least 10% (n = 21) had obtained a COC. An additional 22 respondents had procured COCs for other research, whereas 17 respondents were unaware of them and their purpose. In this era of public scrutiny of medical research, we recommend greater familiarity with and documentation of all human subject protections. PMID- 15345123 TI - Certificates of confidentiality in research: rationale and usage. AB - Certificates of confidentiality (COCs) are a tool to protect researchers from being compelled to release identifying information about their subjects. Whereas institutional review board (IRB) review and informed consent procedures are mandatory tools to protect human subjects, COCs are voluntary. There are limited data about who procures COCs and why, and whether they are useful. Three Institutes of the National Institutes of Health (NIH) provided data on 114 research projects that had received COCs. Eighty-three researchers had procured a single COC and 11 researchers had procured 31 COCs. One hundred and four (91%) of the COCs were obtained by researchers at academic sites, and 17 institutions collectively accounted for 82 COCs. The most commonly cited sources of information about COCs came from colleagues (n = 18, 35%) and previous experience (n = 17, 33%). The most common reasons for procuring a COC were that the research involved genetics (n = 28, 54%), the research could lead to social stigmatization or discrimination (n = 22, 42%), or the research could damage an individual's financial standing, employability, or reputation (n = 21, 40%). These findings show that COCs are often congregated within institutions and by particular individuals. This may be because others are unaware of COCs or because others do not believe they are necessary or useful. PMID- 15345124 TI - Technological advances in stem cell research: developing a consensus. PMID- 15345125 TI - Culture and characterization of human embryonic stem cells. AB - Human embryonic stem (ES) cells offer substantial opportunities for providing well-defined differentiated cells for drug discovery, toxicology, and regenerative medicine, but the development of efficient techniques for their large-scale culture under defined conditions, and for controlling and directing their differentiation, presents a substantial challenge. Markers for defining the undifferentiated cells are well established, based upon previous studies of embryonal carcinoma (EC) cells, their malignant counterparts from teratocarcinomas. These provide valuable tools for monitoring human ES cultures and their state of differentiation. However, current culture techniques are suboptimal and involve the use of poorly defined culture media and the use of feeder cells. Over time, the cells may also acquire karyotypic changes, reflecting genetic selection and adaptation to in vitro culture conditions. Nevertheless, progress is being made. Originally, human ES cells were derived and maintained in medium containing fetal calf serum. They are now widely cultured in a proprietary serum-free formulation (serum replacement from Invitrogen Corp., Carlsbad, CA), and recently we have derived a new human ES line in this medium without fetal calf serum. Human fibroblasts can also be used to replace mouse embryo fibroblasts as feeder cells. We have now found it possible to culture a subline of human ES cells on Matrigel, or purified collagen type IV, laminin, and fibronectin, without feeders or feeder-conditioned medium. These cells nevertheless retain the features of undifferentiated human ES cells, including a capacity for differentiation. Although these cells also carried karyotypic changes, further research focused upon understanding the mechanisms that control self-renewal, apoptosis, and commitment to differentiation will facilitate the development of defined culture conditions that minimize genetic change and optimize the maintenance of the undifferentiated stem cells. PMID- 15345126 TI - Culture and expansion of the human embryonic stem cell line HS181, evaluated in a double-color system. AB - An approach of using RFP-transfected human foreskin fibroblasts (hFS-RFP) to support the growth of GFP expressing human embryonic stem cells (hES; HS181-GFP) is reported. The two-color system was applied to detect interactions between hFS and human embryonic stem cells (hES). After overnight culture, the hES cell colonies showed a behavior of "pushing away" the underlying feeder cells. This phenomenon occurred with both a low and high density of feeders. The density of the feeder cell layer, however, influenced the growth pattern of hES cell colonies. At a high feeder cell density, the hES colonies were more pointed and aligned with the direction of the fibroblasts, whereas less dense feeder layers allowed a more rounded and flat hES colony formation. Not surprisingly, a small fraction of mitotically inactivated feeder cells reattached after passage and remained viable in the cultures for up to four subsequent passages. The prospect of using the two-color system for detection of possible fusion events between hES cells and feeder cells was assessed by screening a large number of cell cultures for double RFP/EGFP expressing cells. The results indicate that fusion events are extremely rare (<10(-6)), or alternatively that after fusion the dual expression of both EGFP and RFP is not easily detected for other reasons. In summary, a two color system allows analysis of colony formation and also helps to identify and follow the differentiation of cells. PMID- 15345127 TI - Endothelial progenitor cell culture for vascular regeneration. AB - A certain population of mononuclear cells in the peripheral blood is capable of contributing to new vessel formation by differentiating into endothelial cells. The basis for native as well as therapeutic neovascularization is not restricted to angiogenesis but includes postnatal vasculogenesis. These cells were discovered by Asahara in 1997 and named endothelial progenitor cells (EPCs). Clinical usefulness of EPCs from human peripheral blood is also suggested in the animal experiments. These results indicate that administering EPCs can be a new clinical strategy for treating ischemic disease, diabetic retinopathy, or neoplasm in which the promotion or inhibition of neovascular formation is critical. However, expansion of EPCs ex vivo is not currently suitable for the clinical setting because of the animal products that are necessary for EPC expansion. To approach the autologous cell-based clinical application of EPCs, we established EPC culture using auto serum. In this article, we will discuss the sources that can generate EPCs and show the usefulness and potential of EPC culture using auto serum in the basic and clinical setting of neovascular formation. PMID- 15345128 TI - Impact of leukapheresis cell composition on immunomagnetic cell selection with the Baxter Isolex 300i device: a statistical analysis. AB - Immunomagnetic cell selection (ICS) of CD34(+) cells is increasingly adopted in allogeneic and autologous transplant settings. Because many variables can affect the final results of ICS, we focused our study toward the influence exerted by the leukapheresis (LKF) cell composition on recovery, purity, and log of T and B depletion of the immunoselected cells. A total of 39 consecutive CD34(+) ICS were performed with the Isolex 300i (Baxter) device on 39 LKF from 9 HLA haploidentical donors and 20 patients. Flow cytometric analysis was performed both on the leukapheresis content and on the immunoselected cells. The statistical analysis was performed utilizing the Pearson's correlation test and the Mann-Whitney U test. The median purity and recovery of the immunoselected CD34(+) cells were 95.3% (IR: 93.0-99.0) and 55.1% (IR: 41.8-68.2), respectively. The median log of T and B depletion were 3.87 (IR: 3.5-4.3) and 2.9 (IR: 2.5 3.5), respectively. Our data indicate that not only the CD34(+) cell load but also the ratio among the cells belonging to the starting fraction can influence the results of ICS. LKF collection protocols have to be addressed to collect an high number of CD34(+) cells (>500 x 10(6)) without taking care of the contaminating cells when the Baxter Isolex 300i device is employed. PMID- 15345129 TI - Comparison of two continuous-flow systems for the collection of peripheral progenitor cells. AB - New advances in apheresis technology allow for the safe and efficient collection of peripheral progenitor cells (PPC). Two blood cell separators were compared with respect to separation results such as PPC yield and contamination of the products. A total of 11 patients (6 multiple myeloma, 4 non-Hodgkin lymphoma, and 1 medulloblastoma) underwent PPC collections with either the Amicus (Baxter) or AS. TEC (Fresenius) blood cell separator. PPC were mobilized by chemotherapy and granulocyte colony-stimulating factor (G-CSF) application. Blood counts were determined before and after apheresis as well as in the PPC product. CD34 antigen expressing cells were measured in the peripheral blood and in the PPC product by flow cytometry. Median baseline CD34 antigen-expressing cells were higher in patients undergoing PPC collection with the Amicus device. More PPC/kg of body weight were collected with this machine (5.3 x 10(6)/kg body weight versus 1.7 x 10(6) in the AS. TEC). The median volume was 129 ml (range 80-156 ml) for Amicus products and 111 ml (range 66-202 ml) for the AS. TEC, respectively. The median platelet contamination of the products from the Amicus blood cell separator was significantly lower than in products from the AS. TEC machine (0.17 x 10(11) versus 0.65 x 10(11), p < 0.001). The data show that a higher yield of PPC was collected with the Amicus machine. The platelet contamination of the products obtained from the two blood cells separators was significantly different. PMID- 15345130 TI - Effect of addition of FLT-3 ligand and megakaryocyte growth and development factor on hemopoietic cells in serum-free conditions. AB - The aim of this study was to clarify the mechanisms that regulate hematopoietic cell expansion in vitro by identifying defined culture conditions. We report the results of experiments with CD34(+) cells from cord blood (CB, n = 13), bone marrow (BM, n = 4), and mobilized peripheral blood stem cells (PBSC, n = 5) using two combinations of cytokines: (A) granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), interleukin-6 (IL-6), stem cell factor (SCF), erythropoietin (EPO), insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (FGF-b) and (B) combination A plus FLT3 ligand (FL) and megakaryocyte growth and development factor (PEG rhMGDF). Cultures of immunoselected CD34(+) cells were performed in serum-free liquid medium without serum substitutes. The area under the curve (AUC) obtained by plotting the logarithm of the total number of viable cells, CD34(+) cells, and CFC per well, toward the week of culture was used as an index of cell expansion. With CB, a significant difference was obtained between the two combinations of cytokines with regard to the total number of viable cells, GM-CFC, and CD34(+) cells. The difference between the two combinations of cytokines obtained with BM was significant with respect to the total number of viable cells and CD34(+) cells but not for the erythroid and myeloid progenitors. When CD34(+) cells from peripheral blood stem cells (PBSC) were cultured in presence of the two combinations of cytokines, the difference in terms of AUC was not statistically significant. Our data indicate additional effects in terms of proliferation and expansion of hematopoietic cells in serum-free conditions when FL and polyethylene glycol (PEG) rhMGDF are included in culture and suggest a differential activity of these cytokines on cells from different hematopoietic sources. PMID- 15345131 TI - Neural differentiation from embryonic stem cells: which way? AB - Embryonic stem (ES) cells can in theory produce all cell types of a living organism while renewing themselves with a stable genetic background. These unique features make ES cells a favorable tool for biomedical researches as well as a potential source for therapeutic application. A first step for approaching to ES cells is the directed differentiation to cells of interest, such as the neural cell lineage. Here, we summarize the up and down sides of each category of neural differentiation protocols that have so far been used in mouse and human ES cells, and introduce an efficient and plausible method used in our laboratory for derivation of neuroectodermal cells from human ES cells. This synthesis has led to our suggestions on issues for future design of neural differentiation protocols. PMID- 15345132 TI - In vitro proliferation and expansion of hematopoietic progenitors present in mobilized peripheral blood from normal subjects and cancer patients. AB - In the present study, we have assessed, in a comparative manner, the in vitro proliferation and expansion potentials of hematopoietic progenitor cells (HPC) present in mobilized peripheral blood from normal subjects (MPB-n; n = 18) and cancer patients (MPB-c; n = 18). The latter included patients with breast cancer (BrCa; n = 8), Hodgkin disease (HD; n = 4), non-Hodgkin lymphoma (NHL; n = 3), and acute myeloid leukemia (AML; n = 3). Progenitor cells from normal bone marrow (BM) and umbilical cord blood (UCB) were included as controls. HPC, enriched by a negative selection procedure, were cultured for 25 days, in serum-free liquid media in the presence of a cytokine combination including early- and late-acting cytokines. Our results demonstrate that the in vitro biological properties of progenitor cells present in MPB differ, depending on whether they are derived from healthy individuals, from patients with solid tumors, or from patients with hematological neoplasias. Among all cell sources analyzed, UCB-derived progenitors showed the greatest proliferation and expansion potentials (1000-fold increase in total cell numbers on day 15, and a 22-fold increase in myeloid progenitor cell numbers, at day 10). Progenitor cells present in MPB from hematologically normal individuals showed proliferation and expansion potentials comparable to those of HPC from normal BM (500-fold increase in total cell numbers on day 15, and a 14-fold increase in myeloid progenitor cell numbers, at day 10). The proliferation/expansion potentials of MPB progenitors from BrCa patients were also within the normal range, although in the lower levels (327 fold increase in total cell numbers, on day 15, and 11.8-fold increase in myeloid progenitors, at day 10). In contrast, progenitors present in MPB from patients with HD, NHL, and especially AML, showed reduced in vitro capacities (119-, 102-, and 51-fold increase in total cell numbers, respectively; and 8-, 4-, and 2.6 fold increase in myeloid progenitor cells, respectively). To our knowledge, this is the first report in which the in vitro proliferation and expansion potentials of HPC from MPB from normal subjects and cancer patients are assessed simultaneously in a comparative manner. PMID- 15345133 TI - Engraftment of splenic tissue as a method to investigate repopulation by hematopoietic cells from host and donor marrow. AB - The lymphohematopoietic function of the spleen in mice varies dependent on age and hematopoietic requirements. A method was developed to study splenic repopulation of mature and progenitor cell populations by grafting neonatal or adult spleen tissue under the renal capsule of splenectomized mice. Two weeks following implant of irradiated syngeneic neonatal spleens into B6-Ly 5.1 or B6 gfp recipients, host lymphoid (B220(+), CD4/8(+)) and myeloid cells (CD11b(+)) had repopulated the splenic grafts and constituted the majority of cells contained in these heterotopic implants. Notably, the percentage of lymphoid and myeloid cells approximated adult levels in contrast to preimplant neonatal spleen levels. This observation indicated relatively rapid repopulation of the grafted tissue by adult host cells and suggests that the repopulation patterns were regulated by the host. Three months post-implantation, the cell composition in the graft remained comparable to adult levels. Microscopic examination demonstrated normal splenic architecture including follicles and red pulp. Lymphocytes within the graft were functional as indicated by their proliferation in response to lipopolysaccharide (LPS) and concanavalin A (ConA) stimulation. Progenitor cell activity determined by colony-forming unit interleukin-3 (CFU-IL 3) levels was also present in these grafts. Splenic implants were then assessed in transplant models following lethal irradiation and syngeneic or allogeneic bone marrow transplantation (BMT). Two weeks post-BMT, adult splenic tissue implants contained donor-derived B cells, T cells, and myeloid cell populations. As typically detected in the host spleen post-BMT, the grafted tissue also contained elevated levels of donor progenitor cells. By 3 months post-BMT, CFU IL3 levels in the graft reflected the decreased levels characteristic of adult levels. The functional integrity of post-transplant splenocytes in the implants was also demonstrated by mitogenic responsiveness. In summary, this method should provide a useful model for the transfer of the splenic microenvironment to study the biology of the spleen in non-transplant and BMT settings. PMID- 15345134 TI - Human embryonal carcinoma stem cells: models of embryonic development in humans. AB - There are few reliable experimental systems available to study the molecular mechanisms that govern human embryonic development. Embryonal carcinoma (EC) cells are pluripotent stem cells derived from teratocarcinomas and are considered the malignant counterparts of human embryonic stem (ES) cells. Several of the existing human EC stem cell lines provide robust and simple culture systems to study certain aspects of cellular differentiation in a manner pertinent to human embryogenesis. Here we review the strategies used to derive and characterize the established and recognized human EC stem cell line TERA2.cl.SP12. Furthermore, we demonstrate the value of human EC stem cells as a model of early development and focus on cell fate determination in the embryonic ectoderm. PMID- 15345135 TI - Generation of neuroprogenitor-like cells from adult mammalian bone marrow stromal cells in vitro. AB - Recently, it has been proposed that bone marrow stromal cells (BMSCs) have a broader capacity for differentiation than previously contemplated. In vitro studies have indicated that BMSCs may have the capacity to differentiate into neuroectodermal-like cells in response to various growth conditions, including those commonly used to maintain and differentiate cultures of primary neural stem cells (NSCs). Interpreting the wealth of data on this subject has been difficult because of variation in the starting cell population and the differences between the methods used to induce their differentiation. Here we evaluate how cultures of expanded BMSCs with a consistent immunophenotype respond to a variety of growth conditions and induction agents and review their ability to form neural like derivatives. In addition, we report on some modifications to previously published techniques for the generation of neural-like cells from BMSCs in vitro. PMID- 15345136 TI - Organized development from human embryonic stem cells after injection into immunodeficient mice. AB - Information concerning the development and differentiation of human embryonic stem (hES) cells in vivo is limited. The present study has focused on the in vivo outcome and differentiation of the hESC line HS181, after injection into SCID/beige mice. hES cell-derived teratomas were explored using histological evaluation and by the identification of markers for differentiated cells and tissues. The analyses identified predominant differentiation along a neuronal lineage, the formation of bone/cartilage and epithelia. Fluorescent in situ hybridization (FISH) analysis with a human-specific probe showed the teratomas to be mainly of human origin, with the most organized areas being exclusively human. Importantly, the study revealed interactions between mouse and human tissues, most notably in the formation of vessels. Both mouse and human cells contributed to specific microstructures in which mouse cells could be observed to take on the appropriate histiotypic appearance. Hence, HS181 cells were able to develop into defined mature tissues, supporting the relevant use of this hES cells model for studies of early human development, given the use of appropriate controls for host contribution. Although extensive mitotic activity implicated progenitor cell activity, no detectable multipotent or malignant areas were observed during the observation period. Persisting undifferentiated hESC were not detected. PMID- 15345137 TI - Mesenchymal stem cells: isolation and therapeutics. AB - Mesenchymal stem cells (MSCs) are progenitors of all connective tissue cells. In adults of multiple vertebrate species, MSCs have been isolated from bone marrow (BM) and other tissues, expanded in culture, and differentiated into several tissue-forming cells such as bone, cartilage, fat, muscle, tendon, liver, kidney, heart, and even brain cells. Recent advances in the practical end of application of MSCs toward regeneration of a human-shaped articular condyle of the synovial joint is one example of their functionality and versatility. The present review not only outlines several approaches relevant to the isolation and therapeutic use of MSCs, but also presents several examples of phenotypic and functional characterization of isolated MSCs and their progeny. PMID- 15345138 TI - The heme oxygenase system: past, present, and future. PMID- 15345139 TI - Role of NF-kappaB and p38 MAP kinase signaling pathways in the lipopolysaccharide dependent activation of heme oxygenase-1 gene expression. AB - Heme oxygenase (HO)-1 is the inducible isoform of the rate-limiting enzyme of heme degradation, which is up-regulated by a host of stress stimuli. The bacterial cell membrane component lipopolysaccharide (LPS) is a prototypical activator of monocytic cells. Here, it is shown that LPS induced the endogenous HO-1 gene expression in RAW264.7 monocytic cells. To investigate the molecular mechanisms of HO-1 gene induction by LPS, we performed transfection experiments with reporter gene constructs containing sequences of the proximal rat HO-1 gene promoter. Deletion and mutation analysis indicated that a cyclic AMP response element/activator protein-1 site (-664/-657), but not an E-box motif (-47/-42), played a major role for LPS-dependent HO-1 gene induction. Up-regulation of HO-1 promoter activity by LPS was decreased by pharmacological nuclear factor-kappaB (NF-kappaB) inhibitors and by cotransfected expression vectors with dominant negative isoforms of NF-kappaB-inducing kinase, inhibitor of NF-kappaB (IkappaB) kinase beta, and IkappaBalpha. Moreover, the p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 and overexpressed dominant negative p38beta decreased, whereas dominant negative p38delta increased, LPS-dependent induction of HO-1 gene expression. The results suggest that the NF-kappaB and p38 MAPK signaling pathways mediate the LPS-dependent induction of HO-1 gene expression via DNA sequences of the proximal promoter region. PMID- 15345140 TI - Ethyl ferulate, a lipophilic polyphenol, induces HO-1 and protects rat neurons against oxidative stress. AB - In the CNS, the heme oxygenase (HO) system has been reported to be active and to operate as a fundamental defensive mechanism for neurons exposed to an oxidant challenge. We have recently shown that both curcumin and caffeic acid phenethyl ester, two phenolic natural compounds, potently induce HO-1 expression and activity in rat astrocytes. We have extended our previous findings examining the effects of two other plant-derived phenolic compounds, with analogous chemical structures, in rat astrocytes and neurons. Ethyl ferulate (ethyl 4-hydroxy-3 methoxycinnamate) (EFE), the naturally occurring ester of ferulic acid, was able to induce HO-1 protein expression. Maximal expression of HO-1 mRNA and protein and a significant increase in HO activity were detected after 6 h of incubation with 15 microM EFE in astrocytes and 5 microM EFE in neurons. Higher concentrations of EFE (50 microM) caused a substantial cytotoxic effect with no change in HO-1 protein expression and activity. Exposure of astrocytes to resveratrol, a phytoalexin derived from grapes, resulted in an increase of HO-1 mRNA, but it was not able to induce HO-1 protein expression and activity. Interestingly, preincubation (12 h) of neurons with EFE resulted in an enhanced cellular resistance to glucose oxidase-mediated oxidative damage; this cytoprotective effect was considerably attenuated by zinc protoporphyrin IX, an inhibitor of HO activity. This study identifies a novel natural compound that could be used for therapeutic purposes as a potent inducer of HO-1 for the protection of brain cells against oxidative and neurodegenerative conditions. PMID- 15345141 TI - Why heme needs to be degraded to iron, biliverdin IXalpha, and carbon monoxide? AB - A large amount of hemoglobin is degraded daily to heme and globin and is replenished by biosynthesis in the bone marrow erythroblasts. "Free heme" can be dissociated from apohemoglobin in vitro and, conversely, native hemoglobin can be renatured from them. Then why does heme need to be degraded to iron, biliverdin IXalpha, and carbon monoxide in vivo? Free heme, i.e., a protein-unbound heme, exists in cells at a very minute concentration and exerts regulatory functions such as the repression of nonspecific delta-aminolevulinate synthase expression and the induction of microsomal heme oxygenase-1 (HO-1). The latter gene expression occurs by way of free heme-mediated derepression of Bach1, a mammalian heme-responsive transcription factor that suppresses the activation of the HO-1 gene. All these events occur at free heme concentrations below 1 microM. In contrast, free heme concentration greater than 1 microM can be toxic because it catalyzes the production of reactive oxygen species. To cope with this problem, the body is equipped with various defense mechanisms against high free heme concentrations. HO is one of the major players in these mechanisms, and it catabolizes free heme to iron, biliverdin IXalpha, and carbon monoxide. These three metabolites of heme by HO reactions have additional important functions and are involved in various critical cellular events. Thus, the breakdown of heme to smaller elements has its own significance in essential cellular metabolism. PMID- 15345142 TI - Bacterial heme oxygenases. AB - The importance of heme oxygenases in heme catabolism, iron utilization, and cellular signaling has been recognized for many years and is a well studied process in eukaryotes. Through the accessibility of an increasing number of bacterial genomes, it has become evident that heme oxygenases are also widespread in prokaryotes. In these organisms, the heme oxygenase reaction serves a similar function as in eukaryotes. A major role of bacterial heme oxygenases has been attributed to acquisition of iron in prokaryotic pathogens, but other functions, such as involvement in phytobilin biosynthesis, have been described. This review summarizes the current state of the art on bacterial heme oxygenase research. The various biological roles of this enzyme in prokaryotes and their biochemical properties will be discussed. PMID- 15345143 TI - Solar ultraviolet A radiation: an oxidizing skin carcinogen that activates heme oxygenase-1. AB - Ultraviolet A (UVA: 320-380 nm) radiation is an oxidizing carcinogen that has proved an ideal agent for demonstrating the oxidant inducibility of the mammalian heme oxygenase-1 (HO-1) gene. The UVA response in cultured human skin fibroblasts and other cell types is mediated by singlet oxygen and is strongly influenced by cellular reducing equivalents. Free heme, an entity that can be generated by UVA irradiation of cells, also appears to be a critical intermediate that can directly influence both the transcriptional activation and repression of the HO-1 gene. Heme release is likely to be of central importance to the inflammatory response in skin and its abrogation by HO. PMID- 15345144 TI - Antioxidant activities of bile pigments. AB - Biliverdin and bilirubin are reducing species and hence potential antioxidants formed by the action of heme oxygenase and biliverdin reductase. Indeed, there is increasing evidence for the suggestion that a beneficial role of the potentially toxic bilirubin may be to act as a powerful chain-breaking antioxidant in biological systems, and that bilirubin may contribute to the cellular and tissue protection seen with increased heme oxygenase. This article reviews the in vitro antioxidant activities of the two bile pigments with emphasis on the different physiological forms of bilirubin and types of oxidants, and discusses these properties in light of the presence and reactivity other nonproteinaceous antioxidants. PMID- 15345145 TI - Alteration in heme oxygenase-1 and nitric oxide synthase-2 gene expression during endotoxemia in cyclooxygenase-2-deficient mice. AB - Sepsis is a systemic inflammatory response to a blood-borne infection that is associated with an extremely high rate of morbidity and mortality. The present article reviews our recent studies involving the role of cyclooxygenase (COX)-2 in host responses to bacterial endotoxemia and its role in the regulation of nitric oxide synthase (NOS)2 and heme oxygenase (HO)-1. COX-2-deficient (-/-) mice display a blunted and delayed induction of the cytokine-inducible genes NOS2 and HO-1 after administration of Escherichia coli lipopolysaccharide (LPS or endotoxin). Translocation and activation of transcription factors important for signaling events during an inflammatory response, such as nuclear factor-kappaB and activating protein-1, are also reduced. In addition, COX-2(-/-) mice have reduced leukocyte infiltration into critical organs (kidneys and lungs) after LPS administration. Interestingly, the absence of COX-2 does not alter the LPS induction of several proinflammatory cytokines in tissue macrophages, but induction of the antiinflammatory cytokine interleukin-10 is exaggerated. After LPS administration, 50% of wild-type (+/+) mice die; however, COX-2(-/-) mice display a dramatic improvement in survival during endotoxemia. Taken together, our findings suggest that COX-2(-/-) mice are resistant to many of the detrimental consequences of endotoxemia. PMID- 15345146 TI - Complex role of heme oxygenase-1 in angiogenesis. AB - Angiogenesis occurring during reparative or pathological processes is driven by various inflammatory mediators that influence the synthesis of growth factors. It has been recognized recently that reactive oxygen species (ROS) and nitric oxide (NO) are important modulators of the synthesis and activity of vascular endothelial growth factor (VEGF), a major angiogenic molecule. Moreover, heme oxygenase-1 (HO-1), a ubiquitous stress-inducible enzyme that is induced by ROS and NO, was recently discovered to be involved in angiogenesis. Genetic overexpression of HO-1 enhanced VEGF synthesis and augmented formation of vascular capillaries, improving the blood flow in ischemic tissues. In addition, by-products of HO-1 exert numerous effects that can also influence angiogenesis in both positive and negative ways. Therefore, the antiinflammatory effects of HO 1 can attenuate the excess formation of blood vessels in inflammatory angiogenesis. In this review, the recent data on the role of HO-1 in angiogenesis are critically discussed. It is suggested that further studies using potent and specific augmentation of HO-1 gene expression by viral vectors, as well as targeted, specific inhibition of HO-1 expression, are required to elucidate fully the complex role of this enzymatic pathway in angiogenesis. PMID- 15345147 TI - Protective role of heme oxygenase-1 in renal ischemia. AB - Oxidative stress, which has been implicated in the pathogenesis of ischemic renal injury, degrades heme proteins, such as cytochrome P450, and causes the elevation in the level of cellular free heme, which can catalyze the formation of reactive oxygen species. Heme oxygenase-1 (HO-1), the rate-limiting enzyme in heme degradation, is induced not only by its substrate, heme, but also by oxidative stress. In various models of oxidative tissue injuries, the induction of HO-1 confers protection on tissues from further damages by removing the prooxidant heme, or by virtue of the antioxidative, antiinflammatory, and/or antiapoptotic actions of one or more of the three products, i.e., carbon monoxide, biliverdin IXalpha, and iron by HO reaction. In contrast, the abrogation of HO-1 induction, or chemical inhibition of HO activity, abolishes its beneficial effect on the protection of tissues from oxidative damages. In this article, we review the protective role of HO-1 in renal ischemic injury, and its potential therapeutic applications. In addition, we summarize recent findings in the regulatory mechanism of ho-1 gene expression. PMID- 15345148 TI - Heme oxygenase and its products in the nervous system. AB - Heme oxygenase (HO) cleaves the tetrapyrrolic ring of cellular heme moieties liberating carbon monoxide (CO) and equimolar amounts of free iron and biliverdin (BV). BV is in turn converted into bilirubin (BR) by the cytosolic enzyme BV reductase. Three HO isoforms have been described to date: HO-1, HO-2, and HO-3. All these isoforms are present in nervous tissue with different localizations and regulation. CO, the gaseous product of HO, exerts its biological effects through the activation of soluble guanylyl cyclase, but alternative signaling pathways, such as the activation of cyclooxygenase, have also been reported in the brain. In vitro and in vivo studies showed that CO, at the hypothalamic level, plays a key role in the modulation of stress response because it inhibits the release of antiinflammatory neuropeptides, such as corticotropin-releasing hormone and arginine vasopressin, and increases body temperature in rodents exposed to psychological stressors (stress fever). In the last few years, a new role of BR as an endogenously produced antioxidant has emerged, and several reports have shown that BR contributes to prevent cell damage mediated by reactive oxygen species, as well as nitric oxide and its congeners. PMID- 15345149 TI - Heme catabolism and heme oxygenase in neurodegenerative disease. AB - Heme oxygenase, the rate-limiting step in heme catabolism, appears to play an important role in a number of neurodegenerative disorders, such as Alzheimer disease. Interestingly, the spatial distribution of heme oxygenase-1 expression in diseased brain is essentially identical to that of the pathological expression of tau, suggesting a key role for both in disease progression. Like heme oxygenase, the expression, phosphorylation, and aggregation of tau are regulated through signal cascades, including the extracellular signal-regulated kinases, whose activities are modulated by oxidative stress. Therefore, the expression of tau and heme oxygenase-1 in a coordinated manner likely plays a pivotal role in the cytoprotection of neuronal cells. This places heme oxygenase at the center of disease pathogenesis and offers a novel therapeutic approach targeted at either the causes or consequences of enzyme induction. PMID- 15345150 TI - Redox regulation in neurodegeneration and longevity: role of the heme oxygenase and HSP70 systems in brain stress tolerance. AB - Efficient functioning of maintenance and repair processes seems to be crucial for both survival and physical quality of life. This is accomplished by a complex network of the so-called longevity assurance processes, which are composed of several genes termed "vitagenes," among these, the heat shock system, a highly conserved mechanism responsible for the preservation and repair of cellular macromolecules, such as proteins, RNAs, and DNA. Recent studies have shown that the heat shock response contributes to establishing a cytoprotective state in a wide variety of human diseases, including ischemia and reperfusion damage, inflammation, cancer, as well as metabolic and neurodegenerative disorders. Recently, the involvement of the heme oxygenase (HO) pathway in antidegenerative mechanisms has received considerable attention, as it has been demonstrated that the expression of HO is closely related to that of amyloid precursor protein. HO induction occurs together with the induction of other heat shock proteins during various physiopathological conditions. The vasoactive molecule carbon monoxide and the potent antioxidant bilirubin, products of HO-catalyzed reaction, represent a protective system potentially active against brain oxidative injury. Given the broad cytoprotective properties of the heat shock response, molecules inducing this defense mechanism appear to be possible candidates for novel cytoprotective strategies. Particularly, manipulation of endogenous cellular defense mechanisms, via the heat shock response, through nutritional antioxidants or pharmacological compounds, may represent an innovative approach to therapeutic intervention in diseases causing tissue damage, such as neurodegeneration. Consistently, by maintaining or recovering the activity of vitagenes, it is feasible to delay the aging process and decrease the occurrence of age-related diseases with resulting prolongation of a healthy life span. PMID- 15345151 TI - Role of oxidative stress in ischemia-reperfusion-induced changes in Na+,K(+) ATPase isoform expression in rat heart. AB - The aim of this study was to assess whether depression of cardiac Na+,K(+)-ATPase activity during ischemia/reperfusion (I/R) is associated with alterations in Na+,K(+)-ATPase isoforms, and if oxidative stress participates in these I/R induced changes. Na+,K(+)-ATPase alpha1, alpha2, alpha3, beta1, beta2, and beta3 isoform contents were measured in isolated rat hearts subjected to I/R (30 min of global ischemia followed by 60 min of reperfusion) in the presence or absence of superoxide dismutase plus catalase (SOD+CAT). Effects of oxidative stress on Na+,K(+)-ATPase isoforms were also examined by perfusing the hearts for 20 min with 300 microM hydrogen peroxide or 2 mM xanthine plus 0.03 U/ml xanthine oxidase (XXO). I/R significantly reduced the protein levels of all alpha and beta isoforms. Treatment of I/R hearts with SOD+CAT preserved the levels of alpha2, alpha3, beta1, beta2, and beta3 isoforms, but not that of the alpha1 isoform. Perfusion of hearts with hydrogen peroxide and XXO depressed all Na+,K(+)-ATPase alpha and beta isoforms, except for alpha1. These results indicate that the I/R induced decrease in Na+,K(+)-ATPase may be due to changes in Na+,K(+)-ATPase isoform expression and that oxidative stress plays a role in this alteration. Antioxidant treatment attenuated the I/R-induced changes in expression of all isoforms except alpha1, which appears to be more resistant to oxidative stress. PMID- 15345152 TI - Regulation of heme oxygenase-1 gene transcription: recent advances and highlights from the International Conference (Uppsala, 2003) on Heme Oxygenase. AB - Recent investigations into the regulation of heme oxygenase-1 gene (hmox-1) transcription have exposed mechanisms of increasing diversity and complexity worthy of a gene whose expression is modulated by a seemingly endless array of physiological, pathophysiological, and nonphysiological agents and conditions. For instance, contrary to initial and prevalent assumptions that inducer dependent gene stimulation is mediated principally by the positive action of transcription activators, it now appears that such induction may occur secondarily to deactivation of the repressor protein, Bach1. As a further complication, heme and cadmium, two potent inducers of the hmox-1 gene, inhibit Bach1 function by different mechanisms-by inhibition of DNA binding or promotion of nuclear export, respectively. Bach1 also plays a role in signal-dependent hmox 1 gene repression, an increasingly appreciated phenomenon that is manifested in a species- and cell-specific manner. Although extreme concentrations of the heme oxygenase-1 protein resulting from the opposing phenomena of gene activation and repression have physiological consequences, even minor modulation in the level of this enzyme, as elicited by variations in the length of a dinucleotide repeat region within the human hmox-1 promoter, may be of clinical relevance. Finally, mechanistic diversity is also apparent in the type and combination of protein kinase-dependent, signal transduction pathways used during hmox-1 gene activation. PMID- 15345153 TI - Long-lasting antioxidant activity in a 600-year-old fermented fruit juice. PMID- 15345154 TI - Experience with seven cases of massive splenomegaly. AB - BACKGROUND: Laparoscopic excision of extremely large spleens has been variously reported, but the usual consensus in the literature is that any patient with a spleen anything over 3000 g is simply not a proper candidate for laparoscopy. This report details our experience with 7 patients (out of 95 operated on) with spleens ranging in size up to 4800 g. METHODS: Our operative procedure involved 3 or 4 trocars placed along a virtually semicircular line centered over the splenic hilum. Splenic attachments were excised with the ultrasonic dissector, and the hilum divided with a stapler. Due to the size of the spleens, Pfannenstiel's incisions were utilized for hand-port placement in the extraction of the specimen. RESULTS: Surgery was successful in all 7 cases, and required no conversion to an open procedure. The average splenic weight was 3450 g (range, 3000-4800 g). Mean operative time was 168 minutes (range, 127-250 minutes). CONCLUSION: Because of improved instrumentation (i.e., laparoscopic stapler and ultrasonic dissector) and refinement of technique, spleens very much larger than what was once considered practicable can now be excised laparoscopically with similarly low morbidity as compared with open splenectomy. PMID- 15345155 TI - Laparoscopic assisted colectomies in kidney transplant recipients with colon cancer. AB - BACKGROUND: Kidney transplant recipients have increased operative risks for major abdominal surgery. The purpose of this study is to present the results of laparoscopic assisted colectomies (LAC) in patients who have received a kidney transplant, and evaluate the difficulty and potential benefits or hazards inherent in this approach. PATIENTS AND METHODS: From September 1993 to March 2003, 820 patients underwent LAC in our service. We studied all patients with kidney transplant and LAC. RESULTS: Three kidney transplantation recipients were included. Two patients were female and one male. The mean age was 65 years (range, 54-73 years). The average time elapsed since transplantation was 8 years (range, 6-10 years), and no patient had experienced problems with rejection. All patients had colon cancer. All of the allografts were contralateral to the side of the colon resection. The mean operative time was 103 minutes (range, 100-105 minutes). There were no complications, renal function remained intact, and there was no need to stop immunosuppression. The average length of hospital stay was 5 days (range, 4-7 days). The mean followup time has been 17 months (range, 3-40 months). Since surgery there have been no episodes of rejection and the patients have been free of cancer. CONCLUSION: The benefits of minimal access surgery seem to be shared by kidney transplant recipients. A key feature may be to avoid stopping immunosuppression perioperatively, therefore lowering the potential risk of rejection. Also, lessening the number of wound-related problems appears important for these patients. LAC in experienced hands must be considered a safe alternative for elective colon resections in highly selected patients with kidney transplants. PMID- 15345156 TI - Increased tumor growth after high pressure pneumoperitoneum with helium and air. AB - BACKGROUND: Tumor growth appears proportional to the pressure of carbon dioxide insufflation during laparoscopic surgery. Air and helium are alternative insufflation gases. The objective of this study is to assess tumor growth after air and helium insufflation at different pressures. METHOD: Ninety-six WAG rats were allocated to either air or helium. In both arms, rats were randomly exposed to a one hour gasless procedure, or to 4 mm Hg, 10 mm Hg, or 16 mm Hg insufflation. At the start of the procedure, 500,000 CC531 tumor cells were injected intraperitoneally. After three weeks, intraperitoneal tumor growth was assessed. RESULTS: Higher insufflation pressures were associated with greater tumor growth. No difference of tumor growth between air and helium insufflation was found. CONCLUSION: In this experimental model, insufflation pressure appeared to have a greater impact on tumor growth than the type of gas. Further studies are necessary but it seems prudent to recommend employment of lower insufflation pressures in laparoscopic oncologic surgery. PMID- 15345157 TI - A prospective randomized trial comparing 2-octyl cyanoacrylate to conventional suturing in closure of laparoscopic cholecystectomy incisions. AB - OBJECTIVE: To compare 2-octyl cyanoacrylate (Dermabond) with conventional suturing for closure of laparoscopic cholecystectomy (LC) wounds in a prospective randomized controlled trial. SUBJECTS: Twenty-five consecutive patients underwent LC at a teaching hospital in the United Kingdom. Patients were randomly allocated to have skin closed with 2-octyl cyanoacrylate or absorbable suture. METHODS: Fifty-one wounds underwent skin closure with absorbable subcuticular suture (3/0 polydiaxanone) and 48 wounds were closed with tissue adhesive. The time to close the wounds, including the placement of dressings, was recorded. At 6 to 8 weeks, the incisions were evaluated with the Hollander wound evaluation scale (HWES). Using a visual analogue scale (VAS), the wounds were also rated by a plastic surgeon who was blinded to the method of closure. RESULTS: The wounds were closed significantly faster in the Dermabond group (mean 165 seconds versus 356 seconds, P = 0.03). There were no differences in the percentage of wounds achieving optimal scores on the HWES (suture 64.7% versus tissue adhesive 60.4%, P = 0.42) nor on the mean VAS (suture 62 mm versus tissue adhesive 59 mm, P = 0.45). CONCLUSION: The tissue adhesive 2-octyl cyanoacrylate is a safe and fast method for closure of LC wounds, with cosmetic results comparable to suturing. PMID- 15345158 TI - Prophylactic antiemetics for laparoscopic cholecystectomy: droperidol, metoclopramide, and droperidol plus metoclopramide. AB - BACKGROUND: Postoperative nausea and vomiting (PONV) is one of the most significant problems in laparoscopic surgery. The antiemetic effects of metoclopramide and droperidol used alone or in combination for prevention of PONV after laparoscopic cholecystectomy (LC) were assessed in this prospective, double blind, placebo controlled randomized study. PATIENTS AND METHODS: A series of 140 patients, ASA physical status I or II, were included in the study. Patients were randomized to one of the following groups: 1, placebo; 2, metoclopramide 10 mg after the induction of anesthesia and placebo at 12 h postoperatively; 3, droperidol 1.25 mg after the induction of anesthesia and droperidol 1.25 mg at 12 h postoperatively; and 4, droperidol 1.25 mg plus metoclopramide 10 mg after the induction of anesthesia and droperidol 1.25 mg at 12 h postoperatively. Patients were observed for 24 hours for PONV, pain, need for rescue analgesics, and adverse events. RESULTS: Data were analyzed using the Student's t-test and chi square test, with P < 0.05 considered statistically significant. The mean incidence of PONV was 54% with placebo, 42% with metoclopramide, 14% with two doses of droperidol alone, and 11% with a combination of metoclopramide plus droperidol. The patients receiving a combination of metoclopramide and droperidol had a significantly lower rate of PONV than those administered metoclopramide alone (P < 0.05) or placebo (P < 0.001). Those receiving two-dose droperidol alone also had a significantly lower incidence of PONV compared with metoclopramide (P < 0.05) and placebo (P < 0.001). There was no statistically significant difference between the metoclopramide and placebo groups. Sedation was significantly greater in patients administered droperidol 12 h postoperatively. CONCLUSION: The combination of metoclopramide and droperidol, and two-dose droperidol alone, were found to significantly decrease the incidence of PONV after LC, whereas metoclopramide alone proved inefficient. PMID- 15345159 TI - Effects of intraperitoneal and extraperitoneal carbon dioxide insufflation on blood gases during the perioperative period. AB - The aim of this study was to evaluate the effects of intraperitoneal and extraperitoneal CO2 insufflation on blood gases during and after laparoscopic surgery. Forty patients were included in this study. Twenty patients underwent elective laparoscopic cholecystectomy with intraperitoneal insufflation (intraperitoneal group) and 20 patients underwent laparoscopic inguinal hernia repair with extraperitoneal insufflation (extraperitoneal group). Arterial blood gases were analyzed at four points: 10 minutes after induction, 10 minutes after insufflation, 10 minutes after desufflation, and 30 minutes after the operation in the recovery room. PaCO2 values in the intraperitoneal group at the four points were 36.8 +/- 4, 39.6 +/- 5.9, 40.7 +/- 4.4, and 42.3 +/- 4.8 mm Hg; in the extraperitoneal group, 35.8 +/- 3.9, 37.4 +/- 4, 42.8 +/- 6.6, and 46.2 +/- 5.9 mm Hg. In the extraperitoneal group, there was a significant increase in postoperative PaCO2 compared to the desufflation PaCO2. In our study, extraperitoneal CO2 insufflation caused increases in PaCO2 values that started perioperatively and continued in the postoperative period. PMID- 15345161 TI - Laparoscopic management of the wandering spleen. AB - Wandering spleen is a rare clinical condition associated with a high incidence of splenic torsion and infarction. The preferred treatment for this condition is splenopexy to reposition the spleen in the left upper quadrant of the abdomen and to preserve splenic function. We present two cases of wandering spleen managed by laparoscopic splenopexy. Both young women presented with intermittent abdominal pain. A CT scan was diagnostic of wandering spleen in each case. Both patients were successfully treated with the laparoscopic splenopexy. The spleen was inserted in a Vicryl mesh bag and fixed in the left upper quadrant in both cases. Both patients had resolution of their preoperative symptoms. Laparoscopic splenopexy has many advantages over open splenopexy or splenectomy, including preserving splenic function, minimal postoperative pain, early discharge from the hospital, and rapid recovery. PMID- 15345160 TI - Endoscopic intraluminal suture plication of the gastric pouch and stoma in postoperative Roux-en-Y gastric bypass patients. AB - BACKGROUND: Endoscopic intraluminal suturing is currently used to treat gastroesophageal reflux disease. This new field of intraluminal gastric surgery may benefit postoperative Roux-en-Y gastric bypass patients. While gastric bypass is highly successful in the majority of patients, significant weight regain can occur over time due to stretching of the gastric pouch and stoma. METHODS: Between November 2002 and January 2003, four patients who previously underwent gastric bypass (GBP) surgery presented with dilated gastrojejunostomy (GJ) anastomosis and weight regain. They gave their consent and were taken to the operating room for upper endoscopy, where they were placed under general anesthesia. Using a flexible endoscopic suturing device with a standard 11-mm endoscope, the dilated GJ anastomosis was plicated to reduce its size, and in two of the patients the gastric pouch was also plicated. RESULTS: Successful stomal plication was performed on all four patients to narrow their dilated stomas that measured > 2 cm preoperatively to < 15 mm postoperatively. Patients were told to go on a puree diet for 4 weeks. One patient had a repeat procedure due to rupture of one suture after eating solid food two weeks after the surgery. The stoma was then plicated with 3 sutures. Two patients had their gastric pouch plicated near the stoma. All patients to date report feeling full earlier with decrease caloric consumption and subsequent weight loss. CONCLUSION: Upper endoscopic intraluminal suturing represents a new field of emerging technology that will certainly find its role in the postoperative bariatric patient. Both the gastric pouch and stoma are within reach for endoscopic intraluminal therapy. How it can aid our patients is currently being studied. PMID- 15345162 TI - Is laparoscopic enucleation the gold standard in selected cases with insulinoma? AB - We present the case of a 49-year-old male with the diagnosis of pancreatic insulinoma who underwent laparoscopic enucleation. The surgical procedure was completed successfully. The patient suffered from postoperative low-volume pancreatic fistula, which was self-limited. This was the first laparoscopic enucleation of an insulinoma in Turkey. The laparoscopic enucleation procedure is an appropriate method for superficially located pancreatic corpus and tail insulinomas. PMID- 15345163 TI - Laparoscopic partial adrenalectomy for recurrent pheochromocytoma in a boy with Von Hippel-Lindau disease. AB - We describe a case of a boy with Von Hippel-Lindau disease who presented with recurrent right adrenal pheochromocytoma 4.5 years after laparoscopic bilateral partial adrenalectomy. The boy had a second laparoscopic adrenal-sparing removal of the tumor. By this technique, not only the recurrent tumor was successfully removed but also the unaffected adrenal cortex could be preserved for the second time. To our knowledge, this is the first published case of its type. PMID- 15345164 TI - Endoscopic management of a retroperitoneal hydatid cyst. AB - Hydatid cyst is a significant health problem in endemic regions. Although progress has been made in medical treatment and interventional radiology techniques, surgery is currently the treatment of choice. The hydatid cysts are known to occur at several unusual sites in the body. With increasing experience in laparoscopy and retroperitoneoscopy, attempts have been made to offer the advantages of minimal access procedures to hydatid disease patients. We present a case report of such a patient whose retroperitoneal hydatid cyst was removed endoscopically. PMID- 15345165 TI - Laparoscopic removal of a retained surgical instrument. AB - We report the case of a patient who underwent laparoscopic removal of a retained surgical spatula two months after a total abdominal hysterectomy and bilateral salpingo-oophorectomy for endometrial carcinoma. The foreign body was detected on a routine radiograph while the patient was been prepared for radiotherapy. Laparoscopy allowed us to explore the entire abdominal cavity for any potential complications of the retained instrument, and the procedure was completed successfully. This case demonstrates the feasibility of using a minimally invasive technique in the removal of a retained foreign body. PMID- 15345166 TI - Endoscopic treatment of gastroesophageal reflux. AB - The treatment of gastroesophageal reflux (GERD) has improved dramatically in recent years. The new endoluminal treatments may bring a relatively simple, minimally invasive therapy that can be simultaneously applied in patients with GERD undergoing diagnostic upper endoscopy. Although the initial results are promising, these new techniques are generating some controversy and require further evaluation with randomized studies and longer follow-ups to define their precise role in the long-term management of the gastroesophageal reflux disease. PMID- 15345167 TI - A simple technique to fix the reservoir in laparoscopic gastric banding. AB - Laparoscopic gastric banding is an established approach for the surgical correction of morbid obesity. Fixation of the reservoir to the anterior abdominal wall is a cumbersome procedure that invariably involves an extension of the skin incision and rather tedious suturing in the depth of the wound. Inadequate completion of this step may lead to tilting of the reservoir, which may hinder future band inflations. This paper describes a simple technique to overcome this difficulty. PMID- 15345168 TI - Ethics in reporting: truth and the scientific literature. PMID- 15345169 TI - Photobiomodulation improves cutaneous wound healing in an animal model of type II diabetes. AB - OBJECTIVE: We investigated the effects of photobiomodulation (PBM) on cutaneous wound healing in an animal model of type II diabetes, Psammomys obesus (Sand Rats). BACKGROUND DATA: 632-nm light has been established as the most effective wavelength for treatment of cutaneous wounds; however, the inconsistent efficacy of PBM may be due to inadequate treatment parameter selection. METHODS: Using 632 nm light, an initial series of experiments were done to establish optimal treatment parameters for this model. Following creation of bilateral full thickness skin wounds, non-diabetic Sand Rats were treated with PBM of differing dosages. Wound healing was assessed according to wound closure and histological characteristics of healing. Optimal treatment parameters were then used to treat type II diabetic Sand Rats while a diabetic control group received no irradiation. In order to elucidate the mechanism behind an improvement in wound healing, expression of basic fibroblast growth factor (bFGF) was assessed. RESULTS: Significant improvement in wound healing histology and wound closure were found following treatment with 4 J/cm(2) (16 mW, 250-sec treatments for 4 consecutive days; p < 0.05). The 4 J/cm(2) dosage significantly improved histology and closure of wounds in the diabetic group in comparison to the non irradiated diabetic group. Quantitative analysis of bFGF expression at 36 h post injury revealed a threefold increase in the diabetic and non-diabetic Sand Rats after PBM. CONCLUSIONS: The results demonstrate that PBM at an energy density of 4 J/cm(2) is effective in improving the healing of cutaneous wounds in an animal model of type II diabetes, suggesting that PBM (632 nm, 4 J/cm(2)) would be effective in treating chronic cutaneous wounds in diabetic patients. PMID- 15345170 TI - Thermographical and morphological studies of Er,Cr:YSGG laser irradiation on root canal walls. AB - OBJECTIVE: This study was performed to investigate the temperature rises on root surfaces and morphological changes on root canal walls of extracted human teeth after irradiation by erbium, chromium: yttrium, scandium, gallium, garnet (Er, Cr:YSGG) laser and to evaluate the efficiency in removing smear layer and debris from the prepared root canal walls in vitro. BACKGROUND DATA: No report on Er, Cr:YSGG laser effects under various conditions and specific tips for endodontic treatments has been reported. METHODS: Sixty extracted human teeth with single and straight root were used in this study. The teeth were prepared at 1 mm short of the apical foramen by a conventional technique using K-files. Fifty-four specimens were irradiated by Er, Cr:YSGG laser at the output powers of 2 W, 3 W, and 5 W for 7 sec with three fiber tips (200, 320, and 400 microm diameters). Temperature changes were recorded thermographically. All teeth were examined by light microscopy or scanning electron microscopy (SEM). RESULTS: Thermographic study showed that the average temperature rises on root surfaces were less than 8 degrees C. Observation by light microscopy revealed the ablation at the apical stop, and that by SEM indicated that laser irradiation at 5 W using a fiber tip with 400 microm diameter was efficient for removing smear layer and debris without carbonization or melting. CONCLUSIONS: The results of this study suggested that the temperature rises during Er, Cr:YSGG laser irradiation at the parameters used in this study are minimal to cause the damage on periodontal and bone tissues. Moreover, it was suggested that it is efficient to remove smear layer and debris without causing any carbonization and melting. PMID- 15345171 TI - Laser-assisted end-to-end vascular anastomosis of elastin heterograft to carotid artery with an albumin stent in vivo. AB - BACKGROUND DATA: Laser-assisted end-to-end vascular anastomosis of an elastin heterograft to native artery may avoid the problems associated with currently available vascular synthetic grafts and conventional suture anastomosis. METHODS: A total of 21 anastomoses in the carotid arteries of seven domestic pigs were performed with an 800-nm laser and an albumin stent plus solder. There were five artery to artery and 16 elastin heterograft to native artery anastomoses. Operative parameters, vascular patency, and histology of the anastomoses were evaluated. RESULTS: Out of 21 anastomoses, 20 were patent at 3 h. The average amount of total energy used was 212 Joules in artery to artery anastomosis and 273 Joules in elastin heterograft to native artery. Histology shows coagulative necrosis of the adventitia, hypereosinophilic contraction band in the media in native artery, and no change in elastin heterografts. CONCLUSIONS: Laser-assisted vascular anastomosis of elastin heterograft to medium size vessel is possible. Albumin stent played an important role in strength of the anastomosis. Chronic studies are warranted to determine long-term patency and histology of the laser assisted vascular anastomosis. PMID- 15345172 TI - Nd:YAG laser influence on microleakage of class V composite restoration. AB - OBJECTIVE: The purpose of this research was to evaluate Nd:YAG laser influence on microleakage of class V composite restorations. BACKGROUND DATA: There are few studies concerning laser with restorative materials, considering microleakage. One more sent. METHODS: Thirty six cavities were prepared and divided into three groups (n = 12): group 1, control; group 2, Nd:YAG laser irradiation before adhesive technique; group 3, Nd:YAG laser irradiation after adhesive technique was used as the single bottle. Adhesive system Nd:YAG laser parameters were 320 microm of fiberoptic; energy/pulse of 40 mJ of energy/pulse; repetition rate of 15 Hz; power of 0.6 W; pulsed and non-contact, 1 mm from the surface; 30 seconds in scanning mode; energy density was 49.76 J/cm(2). Cavities were restored with microhybrid composite. After having used a polishing technique, thermocycling and impermeabilization procedures were performed and specimens were submitted to a microleakage test consisting of immersion in aqueous solution of 50% silver nitrate for 8 h, in the total absence of light. After washing and drying procedures, teeth were embedded to facilitate buccal-lingual vertical sectioning. Microleakage was revealed by light. RESULTS: We accomplished Kruskal-Wallis test at 5% level of confidence and observed no statistically significant difference between the tested groups. CONCLUSIONS: Based on the obtained results, it was possible to conclude that Nd:YAG laser in the parameters used, has no influence on marginal microleakage in composite restorations, independent of the moment the laser was used. PMID- 15345173 TI - Comparison of laser, dry needling, and placebo laser treatments in myofascial pain syndrome. AB - OBJECTIVE: We aimed to evaluate the effectiveness of laser therapy in myofascial pain syndrome treatment. BACKGROUND DATA: Myofascial pain syndrome is a disease that is characterized by hypersensitive points called trigger points found in one or more muscles and/or connective tissues. It can cause pain, muscle spasm, sensitivity, stiffness, weakness, limitation of range of motion and rarely autonomic dysfunction. Physical therapy modalities and exercise are used in the treatment of this frequently encountered disease. METHODS: The placebo controlled, prospective, long-term follow up study was planned with 60 patients who had trigger points in their upper trapezius muscles. The patients were divided into three groups randomly. Stretching exercises were taught to each group and they were asked to exercise at home. Treatment duration was 4 weeks. Placebo laser was applied to group 1, dry needling to group 2 and laser to group 3. He-Ne laser was applied to three trigger points in the upper trapezius muscles on both sides with 632.8 nm. The patients were assessed at before, post treatment, and 6 months after-treatment for pain, cervical range of motion and functional status. RESULTS: We observed a significant decrease in pain at rest, at activity, and increase in pain threshold in the laser group compared to other groups. Improvement according to Nottingham Health Profile gave the superiority of the laser treatment. However, those differences among the groups were not observed at 6-month follow up. CONCLUSIONS: Laser therapy could be useful as a treatment modality in myofascial pain syndrome because of its noninvasiveness, ease, and short-term application. PMID- 15345174 TI - The caries-preventive potential of subablative Er:YAG and Er:YSGG laser radiation in an intraoral model: a pilot study. AB - OBJECTIVE: The aim of the present study was to investigate the effect of sub ablative Er:YAG and Er:YSGG laser radiation on the demineralization of human dental enamel in situ. METHODS: Eighteen enamel specimens were prepared to this end, nine of which were irradiated at 8 J/cm(2) with the Er:YSGG laser and nine at 6 J/cm(2) with the Er:YAG laser (125 pulses per surface). Each test surface was assigned a control surface on the same specimen. Three healthy volunteers from each group subsequently wore three enamel specimens in situ for a period of 1 week. The demineralization was assessed by determining the surface microhardness according to Knoop (KH) before and after wearing. RESULTS: Before wearing, the mean surface hardness measured on the control surfaces was 233 (SD 99) KH (CYSGG) and 162 (SD 59) KH (CYAG). After sub-ablative laser irradiation, the surface hardness values measured were already found to be lower compared to the untreated control surfaces (YSGG: 195 [SD 110] KH/YAG: 112 [SD 72] KH). Marked demineralization was observed on the control surfaces after 1 week of wearing in situ (CYSGG*: 60 [SD 57] KH; CYAG*: 53 [SD 9] KH). After wearing, the hardness on the irradiated test surfaces was ErYSGG*: 133 [SD 58] KH and ErYAG*: 89 [SD 28] KH, and was thus higher than on the control surfaces. CONCLUSIONS: The results indicate a tendency towards increased caries resistance following sub ablative erbium laser irradiation. However, in an analysis of variance model with repeated measures, the tendency in this study failed to reach statistical significance (alpha = 0.05). PMID- 15345175 TI - Circumcision with the Nd:YAG laser contact technique compared with conventional surgery. AB - OBJECTIVE: The present study was undertaken to assess the degree of efficacy of the Nd:YAG laser contact technique as compared with conventional surgery in circumcision. BACKGROUND DATA: Neodymium:yttrium-aluminum-garnet (Nd:YAG) laser in urology has many advantages over conventional surgical techniques. Clinical comparative studies between the Nd:YAG laser and conventional circumcision in boys have not yet been reported. METHODS: This study compared the clinical effects of the Nd:YAG laser contact technique with those of a conventional technique on the grounds of certain perioperative parameters, including operative time, length of hospital stay, postoperative complications and morbidity. Seventy five patients undergoing circumcision were reviewed retrospectively. Operative time, length of hospital stay, and postoperative morbidity were analyzed. Differences between contact Nd:YAG laser surgery and conventional surgery were compared. RESULTS: Operative time in the Nd:YAG laser treated group was significantly increased compared with conventionally treated patients (31.6 +/- 3.6 min vs. 19.8 +/- 1.9 min, p < 0.001). The length of hospital stay and early and late postoperative morbidity were significantly decreased in circumcised patients with the Nd:YAG laser contact technique compared with the conventional technique regardless of the indication for surgery (9.71 +/- 4.5 h vs. 11.8 +/- 5.1 h, p < 0.05 and 2% vs. 9%, p < 0.05 and 0% vs. 7.2%, p < 0.05, respectively). CONCLUSIONS: These results indicate that the Nd:YAG laser contact technique is an effective laser-assisted procedure alternative to the conventional technique in circumcision with virtually no significant postoperative morbidity. PMID- 15345176 TI - The efficacy of low-power lasers in tissue repair and pain control: a meta analysis study. AB - OBJECTIVE: We used statistical meta-analysis to determine the overall treatment effects of laser phototherapy on tissue repair and pain relief. BACKGROUND DATA: Low-power laser devices were first used as a form of therapy more than 30 years ago. However, their efficacy in reducing pain or promoting tissue repair remains questionable. METHODS: Following a literature search, studies meeting our inclusion criteria were identified and coded. Then, the effect size of laser treatment, that is, Cohen's d, was calculated from each study using standard meta analysis procedures. RESULTS: Thirty-four peer-reviewed papers on tissue repair met our inclusion criteria and were used to calculate 46 treatment effect sizes. Nine peer-reviewed papers on pain control met the inclusion criteria and were used to calculate nine effect sizes. Meta-analysis revealed a positive effect of laser phototherapy on tissue repair (d = +1.81; n = 46) and pain control (d = +1.11; n = 9). The positive effect of treatment on specific indices of tissue repair was evident in the treatment effect sizes determined as follows: collagen formation (d = +2.78), rate of healing (d = +1.57), tensile strength (d = +2.13), time needed for wound closure (d = +0.76), tensile stress (d = +2.65), number and rate of degranulation of mast cells (d = +1.87), and flap survival (d = +1.95). Further, analysis revealed the positive effects of various wavelengths of laser light on tissue repair, with 632.8 nm having the highest treatment effect (d = +2.44) and 780 nm the least (d = 0.60). The overall treatment effect for pain control was positive as well (d = +1.11). The fail-safe number-that is, the number of studies in which laser phototherapy has negative or no effect-needed to nullify the overall outcome of this analysis was 370 for tissue repair and 41 for pain control. CONCLUSIONS: These findings mandate the conclusion that laser phototherapy is a highly effective therapeutic armamentarium for tissue repair and pain relief. PMID- 15345177 TI - Structural changes of Er:YAG laser-irradiated human dentin. AB - OBJECTIVE: The aim of this study was to investigate the phase, compositional, and morphological changes of Er:YAG laser-irradiated dentin. BACKGROUND DATA: To date, nothing comprehensive has been reported about the phase and compositional changes of human dentin after Er:YAG laser irradiation. METHODS: The human dentin was irradiated by Er:YAG laser with irradiation energies from 300 mJ/pulse-10 pps 10 sec to 700 mJ/pulse-10 pps-10 sec with and without water spray. After irradiation, the specimens were analyzed by means of x-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). RESULTS: After exposure to Er:YAG laser, dentin showed no phase changes on the x-ray diffractometer. In Fourier transform infrared analysis, two absorption bands at 2200 and 2015 cm(-1) could be traced on dentin treated by Er:YAG laser with irradiation energies beyond 300 mJ/pulse-10 pps-10 sec and without water spray. The OH(-) band disappeared at 1630 cm(-1). Scanning electron micrographs revealed that laser energy of 500 mJ/pulse-10 pps-10 sec was sufficient to prompt melting and recrystallization of dentin crystals. CONCLUSIONS: Our study demonstrated that Er:YAG laser irradiation on dentin with water spray would not significantly change the structure and composition. Therefore, water cooling is important for reducing the thermal effect of Er:YAG laser. PMID- 15345178 TI - Effects on periradicular periodontal tissues of root canal irradiation with Er:YAG laser in rats. AB - OBJECTIVE: The effects of Er:YAG laser irradiation on the periodontal region during root canal preparation were evaluated using rats histopathologically. BACKGROUND DATA: The effects on periodontal tissues along the root surface and apical area during root canal preparation using Er:YAG laser irradiation in vivo are not clear. METHODS: One hundred mesial root canals of mandibular first molars in rats were divided into four groups. In three of these groups, root canals were irradiated using an Er:YAG laser at 2 Hz and 34, 68, and 102 mJ/pulse for 30 sec. Non-irradiated canals served as controls. The effects of laser irradiation on the periodontal tissues along the root surface and apical area were evaluated at 0, 2 days, 1, 2, and 4 weeks after irradiation under light microscopy. RESULTS: At 4 weeks after treatment, no inflammation or resorption was observed in any cases in the control or 34 mJ/pulse-irradiated groups. However, moderate to severe inflammation with resorption of root surface was observed in four of five cases (80%) in the 102 mJ/pulse-irradiated group, which was significantly different from the control (p < 0.01). CONCLUSIONS: These results suggest that the effects on periodontal tissues during root canal preparation by Er:YAG laser irradiation after pulpectomy are minimal, if appropriate parameters are selected, and this is a potential therapy for human root canals of teeth. PMID- 15345179 TI - Histological and TEM examination of early stages of bone healing after Er:YAG laser irradiation. AB - OBJECTIVE: The aim of this study was to analyze the early healing process of bone tissue irradiated by Er:YAG laser and compare it with that treated by mechanical drilling and CO(2) laser. BACKGROUND DATA: Er:YAG laser has a great potential for cutting hard tissues as it is capable of ablation with less thermal damage. METHODS: Twenty-four male Wistar rats were used for this study. The calvarial bone of rats was exposed and straight grooves were prepared by Er:YAG laser, mechanical bur and continuous wave CO(2) laser. Four rats each were sacrificed at six time points: 10 min, 6 and 24 h and 3, 7, and 14 days post-surgery. Sections were prepared for light and transmission electron microscopic (TEM) observations. RESULTS: Compared to mechanical bur and CO(2) groups, the inflammatory cell infiltration adjacent to the irradiated bone surface, fibroblastic reaction, and revascularization were more pronounced in the Er:YAG laser-irradiated tissues. A cell-rich granulation tissue with fibroblasts and osteoblasts was predominant in 7-day specimens of Er:YAG laser group. Histopathological analysis of 14-day specimens in the Er:YAG group also revealed significantly greater new bone formation, compared with the mechanical bur and CO(2) laser groups. CONCLUSIONS: Initial bone healing following Er:YAG laser irradiation occurred faster than that after mechanical bur and CO(2) laser. Er:YAG laser treatment may be advantageous for wound healing of bone tissue, presumably by providing a favorable surface for cell attachment. PMID- 15345180 TI - Tensile bond strength of a one-bottle adhesive system to indirect composites treated with Er:YAG laser, air abrasion, or fluoridric acid. AB - OBJECTIVE: This in vitro study evaluated the tensile bond strength of Single Bond adhesive system to indirect composite surfaces treated with Er:YAG laser, fluoridric acid, or air abrasion. BACKGROUND DATA: Application of adhesive techniques is well established in restorative dentistry, yet the influence of surface treatments on the bond strength of adhesive systems to indirect composite restorations prior to luting or repair procedures remains unclear. METHODS: Two hundred and ten samples made of three indirect composites (BelleGlass, Sculpture, and Targis) were divided into 21 groups (n = 10) and treated with (1) control (no treatment); (2) silane (RelyX 3M ceramic primer); (3) 10% fluoridric acid; (4) Er:YAG laser; (5) Er:YAG laser + 10% fluoridric acid; (6) air abrasion; and (7) air abrasion + 10% fluoridric acid. Afterwards, silane and Single Bond adhesive system were applied on each treated surface, and a direct composite resin (Z250) cone was built. The prepared specimens were submitted to tensile bond strength test and stereoscopy evaluation to determine the type of failure after rupture. RESULTS: Bond strength mean values were statistically different in function of indirect composite and surface treatment. BelleGlass treated with Er:YAG laser had the highest mean value (22.92 MPa), and Sculpture conditioned with fluoridric acid had the lowest mean (11.88 MPa). Stereoscopy showed a predominant cohesive failure in Targis resin and mixed type for Sculpture and BelleGlass. CONCLUSIONS: These results suggest that Single Bond tensile bond strength improves with Er:YAG laser as a surface treatment compared to fluoridric acid or air abrasion. PMID- 15345181 TI - ArF-193 excimer laser and Emdogain in the treatment of experimental periodontitis: an experimental study in rabbits. AB - OBJECTIVE: We aimed to investigate how the progress made on laser technology during the last ten years could overcome this obstacle and allow the use of lasers in periodontology, together with the application of a number of products permitting the regeneration of periodontal tissues. BACKGROUND DATA: The use of lasers in dentistry remains controversial, in spite of their increasing application in medical practice. The main reason for this discrepancy is the frequent report of damage to surrounding tissues and the dental pulp, due to the energy transfer, from the site of laser impact. METHODS: Experimental periodontitis was initiated in fifteen rabbits. Animals were divided into five equal groups. In the control group, no therapy was applied. The remaining four groups were treated with curettage or ArF 193 excimer laser, under conditions of strict control of frequency, fluency, and application, without or with the application of a periodontal healing product (Emdogain). Laser was applied by the use of a new, articulated arm for beam delivery. Pocket depth and microscopic analysis were performed three weeks after treatment. RESULTS: Our results show that all treatment groups decreased pocket depth significantly. ArF193 excimer laser does not produce any histological damage to the dental pulp, and facilitates periodontal regeneration. This result is highly facilitated by the application of Emdogain). CONCLUSIONS: The use of UV lasers, under a tight control of its energy, may be a valuable tool for the treatment of periodontal diseases, especially combined with the use of healing products. Further study is need to confirm these results. PMID- 15345183 TI - Photomedicine and laser literature watch. PMID- 15345184 TI - Work, environment, immune system and human health. PMID- 15345185 TI - Role of the HLA-DP Glu 69 and the TNF-alpha TNF-alpha 2 gene markers in susceptibility to beryllium hypersensitivity. AB - Berylliosis is an environmental chronic inflammatory disorder of the lung caused by inhalation of beryllium dusts, characterized by the accumulation of CD4+ T cells and macrophages in the lower respiratory tract. Beryllium presentation to CD4+ T cells from patients with berylliosis results in T cell activation and these Be-specific CD4+ T cells undergo clonal proliferation and Th1-type cytokine production such as interleukin-2, interferon-gamma and tumor necrosis factor alpha. In exposed workers, genetic susceptibility to this granulomatous disorder is associated with major histocompatibility gene and the TNF-alpha gene. The HLA DP glutamic 69 residue was shown to be the MHC genetic marker associated with disease susceptibility; furthermore the TNF-alpha TNFA-308*2 allele was found to be independently associated with HLA-DP Glu69 in the determination of berylliosis risk. PMID- 15345186 TI - Subclinical neurophysiological effects of manganese in welding workers. AB - High-level occupational manganese (Mn) exposure has been reported to induce irreversible brain alterations determining a Parkinson-like disease. This study aimed to assess subclinical neurophysiological alterations in welding workers. They were employed in a machine building factory with an average Mn exposure <200 mg/m3. Sixty-eight workers (mean age: 34 years; mean Mn exposure duration: 16 years) and 42 flour factory workers (control group) with similar age and smoking habit were recruited. Autonomic nervous function test battery (ANSFT), composed of Valsalva maneuvre-induced heart rate variation (HR-V), heart rate variation following deep breathing (HR-DB) and heart rate variation following immediate standing up (HR-IS) was assessed. Electroencephalogram (EEG), brain electricity activity mapping (BEAM) were also performed. HR-V, HR-DB, and HR-IS were significantly lower in Mn-exposed subjects showing altered autonomic nervous system activity, parasympathetic-sympathetic imbalance and, and consequently, altered cardiovascular regulation and reactivity. The EEG of the Mn-exposed workers evidenced beta-wave rhythms significantly reduced, theta-waves markedly increased and abnormal wave activities of either localized or diffusive type. In the same workers BEAM revealed higher theta, delta and beta power values in the F7 area, lower d power values in the FP1, FP2 and C4 areas as well as dissymmetry in the central area, parietal region and occipital region. This study suggests that Mn impairs neuron activity within central nervous system. In this context, brainstem parasympathetic and sympathetic centers receiving axon projections from cortical and diencephalic areas, may reflect Mn effects on upper pathways. However, direct actions of Mn on these centers cannot be excluded. PMID- 15345187 TI - Neurobehavioral functions, serum prolactin and plasma renin activity of manganese exposed workers. AB - Objective of this study was to assess effects of manganese (Mn) exposure on 56 workers employed in a Mn welding workshop of a machine building factory in Taiyuan (Shanxi Province, P.R. China) for a mean period of 16.1 years. The mean air Mn level in the workshop was 138.4 microg/m3. Neurobehavioral Core Test Battery (NCTB), including the Profile fo Mood States, (POMS), was performed. Blood pressure (BP) increase following immediate stand-up (BP-IS), serum prolactin (PRL) and plasma renin activity (PRA) in supine position were also determine. Most of the NCTB scores of the Mn-exposed workers were lower than those of controls, while the POMS scores were higher, indicating a Mn-induced impairment of neurophysiological functions and a deflection of mood towards negative emotion states. PRL values of the Mn-exposed workers were higher than those of the controls. BP-IS of Mn-exposed workers was significantly lower than that of the controls. PRA of the same workers was augmented more that 200%. In the Mn-exposed workers, the higher PRL values are possibly due to a reduced inhibitory effect on pituitary lactotrope cells by the tubero-infundibular dopamine system; the decreased BP-IS was referred to imbalance between the sympathetic and parasympathetic activities, whereas the higher basal PRA was thought to depend on neuroendocrine changes (including increased central sympathetic tone) and/or on a direct effect of Mn on renal juxta-glomerular cells. On the whole, this study demonstrates that occupational Mn exposure is responsible for neurobehavioral changes coexisting with alterations of neuroendocrine and humoral systems. PMID- 15345188 TI - Study on OELs for enzyme-containing detergent in China. AB - This study is aimed at setting occupational exposure levels for total detergent dust and enzymes in detergent industries. The study population consisted of 795 workers from four enzyme-containing detergent manufacturing plants (A1, A2, B1 and B2), and 156 control workers from an electronic assembly factory. Work environment monitoring was conducted using high volume of air sampler fro measuring the concentration of total dust (mg/m3), and analyzing the level of enzyme (ng/m3) by ELISA method. A standard questionnaires, pulmonary function test, and skin prick test are used to assess health effects. The levels of detergent total dust varied from 0.2 mg/m3 to 12.54 mg/m3. For enzyme levels, in A1, B1 and B2, the concentration ranged from non-detectable to 9.92 ng/m3 and in A2, the concentration was analyzed by enzyme activity methods and was expressed as Gu/m3 (1 Gu/m3 = 16 ng/m3). The concentration is between 0.16-31.36 ng/m3. Non specific irritation rates in exposed workers were significantly higher than that in controls. Based on the data collected from A1, B1 and control plants, 95% benchmark dose lower bound were calculated as 1.17 mg/m3. The difference of pulmonary function between exposed workers and controls is not significant. The results of SPT showed that neither Savinase- nor Alcalase-induced sensitization was found in controls. The prevalence rates of sensitization for Savinase and Alcalase were ranged between 3.2% and 31% in all enzyme-containing detergent manufacturers investigated. No case of occupational asthma was observed. For total dust, 1 mg/m3 is suggested as permissible concentration-time weighted average (PC-TWA), and 2 mg/m3 as permissible concentration-short term exposure limit (PC-STEL). For the enzyme Subtilisins, 15 ng/m3 is suggested as PC-TWA, and 30 ng/m3 as PC-STEL. PMID- 15345189 TI - Are thyroid dysfunctions related to stress or microwave exposure (900 MHz)? AB - In the last decade, numerous scientific evidence suggested possible adverse health effects from exposure to electromagnetic fields (EMF'S) and the use of mobile phones. According to some studies EMF induced changes of trans-membrane Ca++ flux may lead to altered metabolism and/or secretion of neurohormones including TSH, ACTH, GH, prolactin and melatonin. The aim of this research was to analyse the effects of mobile phone use on thyroid function and to evaluate the possible role of occupational stress. 2598 employees (1355 men and 1243 women) with different duties (vendors, operators and network technicians) were included in the study. Exposure to EMF'S, generated by mobile phones, was assessed both by submitting a questionnaire directly to the employees and acquiring data regarding conversation times. The workers were divided into three groups on the basis of their personal mobile phone use. Moreover, a group of 160 workers with TSH values below 0.4 UI/l was characterized. No statistically significant difference regarding TSH values below 0.4 UI/l was observed among workers with different duties but there was a greater prevalence of subjects with low SH values among 192 employees with more than 33 hrs./month conversation time; this difference was statistically significant (p<0.05). On the basis of our data, it is not possible to establish whether this result is determined by exposure to EMF'S from mobile phones of by the stress of using these instruments. PMID- 15345190 TI - Clastogenic but not apoptotic effects on human artery endothelial cells by concentrations of inorganic lead inhibiting their nitric oxide production. AB - Human coronary artery endothelial cells (HCAEC 5156) were cultured as monolayers and exposed to concentrations of lead (as acetate, Pb) in the culture medium similar or lower than those commonly found in the blood of human beings occupationally or environmentally exposed to this element. Only at the concentration of 200 ng/mL, Pb reduced growth rate of HCAEC 5156 cells starting from the 3rd day and up too the 5th day of incubation. On the other hand, Pb (0.2, 2 and 200 ng/mL) increased concentration-dependently micronuclei formation in binucleated HCAEC 5156 cells, as it was shown by the cytokinesis-blocked micronucleus assay (CMBN assay) carried out after 48 hours of exposure to the metal. However Pb was unable, at all the above concentrations to induce apoptosis in the HCAEC 5156 cells following a 48 hour-exposure, as shown by an electorphoretic apoptotic DNA fragmentation test. Moreover, Pb (2 and 200 ng/mL) reduced significantly the concentration of nitric oxide (NO, determined analytically as L-citrulline) in both culture medium and cytosol of HCAEC 5156 cells following a 7 day-exposure to the element. Results were discussed also in relation so evidences of other studies reporting genotoxic and/or apoptotic effects of Pb on various cell types at very elevated dosages of concentrations. The observed clastogenic effects of Pb were explained through a series of mechanisms involving interactions between oxygen reactive species and NO and/or reduced NO synthesis in the endothelium, thus leading to a depressed NO bioavailability. This research first shows that Pb is provided with clastogenic but not apoptotic effects on cultured human endothelial cells. It was emphasized that such effects are induced by Pb concentrations similar to those commonly found in blood and tissues of laboratory animals showing Pb induced cardiovascular and/or neuropsychological alterations. PMID- 15345191 TI - Secretory IgA in saliva and academic stress. AB - Several reports have proposed that the concentration of secretory immunoglobulin A (S-IgA) in saliva is an indicator of psychological stress. With this in mind, we decided to examine it in 10 second year medical student volunteers at Kawasaki Medical School course between May 4 and July 13, 2000 and discussed the relationship between S-IbA and the stress from academic examinations. Saliva was collected three times (on rising, at forenoon, and at bedtime) every Thursday. During this period, sporadic academic examinations were held twice and term end examination occurred during the last two weeks. Results showed the concentration of S-IgA significantly higher at the on rising time-point than at the other two time-points. There was also a tendency for the S-IgA level in saliva to be higher on the day before academic examinations and during them and lower on the days between these examinations. In addition, daily variations in the S-IgA concentration sometimes seemed to be disturbed by other academic stress. Therefore it may be possible to use this measurement to monitor psychological stress in students and workers. PMID- 15345192 TI - Stressful life events of dental students and salivary immunoglobulin A. AB - To study the perceived sources of stressful events in dental students and the relationship between their self-perceived stress levels and salivary IgA. Undergraduates as well as postgraduates at the Faculty of Dentistry, National University of Singapore were surveyed one month after the new term. A 38-item dental environmental stress (DES) questionnaire, with subscales of academic work (AW), clinical factors (CF), faculty and administration factors (FA) and personal factors (PF), was used to identify the potential stressors in the dental environment. A 4-point perceived stress scale was used to rank their self perceived stress levels. Enzyme linked immunosorbent assay method was used to determine the salivary IgA level. One hundred and thirty students (81.3%--valid response rate) participated in the study. Overall, students ranked AW with the highest score (mean 2.76), followed by CF (2.67), FA (2.24) and PF (2.16). Among the 38 items of DES questionnaire, 1st year students perceived "fear of being unable to catch up if behind" as the most stressful event (mean 3.30). For 2nd and 3rd year students, examination and grades had the highest scores (mean 3.28, 3.19, respectively). Completing graduation requirements was the most important stressor for 4th year students (mean 3.89). Post graduates perceived atmosphere created by clinical faculty was most stressful to them (mean 3.05). The mean total perceived stress scores were highest (22.1) in 1st year students and lowest (21.0) in postgraduates, however, no significant different among various classes. First year students had had the lowest IgA secretion rates (geometric mean [GM] 46.8 microg/min), significantly lower (p<0.05) than postgraduates (GM 79.4 microg/min). An inverse correlation was noted between perceived stress scale and log IgA secretion rates (r= -0.20, p= 0.002). AW was also significantly inversely correlated with salivary IgA (r= -0.18, p= 0.04). Dental students in different academic years perceived different important stressors. Salivary IgA secretion rate correlated inversely with self perceived stress. PMID- 15345193 TI - Study of immune alterations in patients with chronic fatigue syndrome with different etiologies. AB - The Chronic Fatigue Syndrome (CFS) is characterized by symptoms lasting for at least six months and accompanied by disabling fatigue. The etiology of CFS is still unclear. At the National Center for Study of the Infectious Diseases Department of the Chieti University some immune investigations were performed with the purpose of detecting markers of the disease. CD4+, CD8+, NK CD56+ and B CD19+ lymphocytes were studied in 92 male and 47 female patients and in 36 control subjects. CFS patients were divided in three groups with a post infectious onset (PI-CFS), an non post-infectious onset (NPI-CFS) and a non post infectious onset with associated infections (NPI-CFS + AI). Both CD4+ and CD8+ lymphocytes were reduced in the CFS patients. However, the CD4+/CD8+ ratio was increased in the CFS patients without difference between males and females. CD56+ cells of CFS patients were also reduced. In particular, blood CD56+ cells counts were significantly higher in PI-CFS patients than in the NPI-CFS subjects. These data confirm our preliminary results suggesting a key-role of a dysfunction of the immune system as a precipitating and-or perpetuating factor of the syndrome. PMID- 15345194 TI - Immune effects of nickel. AB - Data on nickel immunomodulation are contradictory. The most consistent immune effects are suppression of immune responses. It has been shown that T-lymphocytes and NK cells are more susceptible to nickel toxicity than are B lymphocytes or macrophages. Data reported about cytokine production in human and nickel reactive T-cell clones are also conflicting. Some authors studied showed a higher synthesis of IL4, IL5 and IL13 but not of IFN gamma and TNFalpha in Ni allergic subjects. We found that the addiction on NiSo4 to the PBMC cultures of non sensitised subjects induces a reduction of release of IL5, IFN gamma and TNFalpha. Our studies demonstrate a clear difference in the NK cell activity between nickel-tolerant and intolerant individuals. In particular NK cell activity in reduced in sensitised patients respect to the normal subjects and the addition of Ni has immunotoxic potential. Researches are in progress in an Attempt to correlate the present data with other immune parameters and to measure the effects of a Ni Free diet on the immune system of subjects with Ni intolerance. The comprehension of the mechanisms inducing these changes requires further studies in the uptake and intracellular distribution and binding of the metal. PMID- 15345195 TI - Genomic organization and cytokine-mediated inducibility of the human TRIM-8/Gerp gene. AB - Cytokine signaling is negatively regulated by a set of SH2 domain-containing proteins, the Suppressors of Cytokine Signaling (SOCS) acting as intracellular modulators. Experimental evidence indicates that SOCS gene expression is induced by cytokines and pro-inflammatory stimuli and is highly controlled both at transcription and translation level. Furthermore, SOCS proteins appear rapidly degraded inside the cells, mostly controlling their stability by interacting with specific molecules such as elongin B and C. It has been shown that SOCS-1/JAB, a member of the SOCS family, interacts with TRIM-8/Gerp, a new ring protein specifically binding SOCS-1 recombitant polypeptide in-vitro and in-vivo. Trim 8/Gerp, transcribes IFN-gamma in epithelial and lymphoid cells and is expressed mostly ubiquitously in murine and human tissues. Here in this report we present the genomic organization of this new SOCS-1 interactor, and we add new tools for extending investigation of the complex mechanism that undergoes negatively regulation of cytokine signaling. PMID- 15345196 TI - Occupational asthma due to low molecular weight agents. AB - Occupational asthma is defined as variable airflow obstruction and airways hyperresponsiveness caused by exposure to agents present in the workplace. Low molecular weight agents such as isocyanates, aldehydes, anhydrides, colophony, dyes, persulphate, amines, acrylates and metals are steadily increasing as causative agents of occupational asthma. Isocyanates, aldehydes and anhydrides my cause sensitisation through an IgE mediated response in some workers. These agents act as haptens which combine with a carrier protein to form a complete antigen. Assays for the detection of specific IgE are standardized for very few agents and have a good specificity, but poor sensitivity. The diagnosis of occupational asthma relies not only on a suggestive hystory showing that asthma is caused or exacerbated specifically by work exposure, but in most cases needs to be confirmed by objective means. Combined monitoring of lung function parameters, such as peak expiratory flow rate at the work site and non specific bronchial hyperresponsiveness during and away from exposure, is necessary. The "gold standard" for confirming a diagnosis in an individual worker still remains the specific bronchoprovocation test, which has now reached a high degree of sensitivity, specificity and reproducibility for agents such a s isocyanates. In occupation asthma due to low molecular weight agents there are no individual risk factors which could predict the susceptibility to develop the disease. The primary prevention is based on appropriate interventions tn the workplace. The strict medical surveillance of workers may allow the early diagnosis and removal from further exposure in order to prevent morbidity and disability. PMID- 15345197 TI - The relationship of atopy, smoking, and sensitization to methyltetrahydrophthalic anhydride. AB - We investigated the association of smoking, atopy and helper T (Th) cytokines with sensitization to methyltetrahydrophthalic anhydride (MTHPA) in occupationally exposed subjects. A population of 147 workers from two condenser plants using epoxy resin with MTHPA underwent a questionnaire survey and serologic investigation. Total and MTHPA-specific IgE levels were measured by the Pharmacia CAP system, and serum levels of interleukin (IL)-4, IL-13, and interferon-gamma (IFN-g) by enzyme immunoassay. The Pharmacia CAP-Phadiatop test, which detects serum IgE specific to most common aeroallergens, was also used. Ninety-six (65%) of the currently exposed workers had positive MTHPA-specific IgE. A significant difference was found in the frequency of positive specific IgE between atopic and non-atopic subjects (P<0.01), but not between smokers and non smokers. As for smoking, the frequency of positive specific IgE was significantly (P<0.005) higher in smokers than that in non-smokers in non-atopic subjects. Multiple logistic regression analysis also confirmed significant contribution of atopy and smoking to the development of specific IgE (odds ration=3.2 and 10.5, respectively), suggesting that atopic subjects who became sensitized to P<0.01 may become sensitized to common aeroallergens. On the other hand, none of the Th cytokines contributed to the elevation of specific IgE levels. These results suggest that atopic subjects and non-atopic smokers are at increased rist of sensitization by P<0.01. However, to evaluate conclusively the effect of atopy on sensitization, further prospective studies are necessary. PMID- 15345198 TI - Respiratory risks in tunnel construction workers. AB - Respiratory diseases are frequent in tunnel construction workers. A group of 144 subjects randomly selected from the population of 2516 workers engaged in the construction of the railway tunnel under the Appennine Mountains, from Bologna to Firenze was investigated. A group of 69 males comparable for age, living area and habits was studied as a control group. Assessment of air pollutants (NO, NO2, SO2, total dust, silica %) was carried out by means of fixed monitoring stations as well as personal monitors. All the subjects included in the study were examined with a standardised protocol which included physical examination, lung function tests (before and after shift work) and a questionnaire to college respiratory symptoms. Low concentrations of environmental pollutants were evidenced. Significantly lower values of FEV1 and PEF were determined in the worker group pared to controls. A significant decrease in respiratory parameters was shown after shift work. Variables capable of influencing the decrease in parameters include smoking habits, work activity, presence of cough and expectoration, period of the year (spirometries resulted worse in the winter time). Significantly lower values of FEV1 and PEF were evidenced in the workers compared to controls. In spite of the present low work environmental exposure conditions, some physiologic parameters appear altered in tunnel construction workers. This may depend on a variety of noxious agents present in the working environment. PMID- 15345199 TI - Altered cytokine production in mice exposed to lead acetate. AB - Previous investigations have shown that Pb exerts immunotoxic effects. Object of this study were Th1 and Th2-type immune responses of mice to Pb exposure. Adult Swiss male mice were administered 0, 40 and 400 mg/l of Pb (as acetate) in drinking water for 14 days. At the end of the treatment, blood Pb was determined and two Th1 cytokines (IL-2, INF-gamma) and one Th2 serum cytokine (IL-4) were measured. A significant increase in IL-4 production was observed in the mice exposed to 40 mg/l of Pb, while a further increase in IL-4 production was associated with a decrease in INF-gamma production in mice exposed to 400 mg/l of Pb. On the other hand, Pb exposure did not induce changes of serum IL-2 (involved also in the Th0 immune pattern). Our findings indicate that low level Pb exposure enhances a Th2 response. A high Pb does can either stimulate the Th2 immune activity or reduce the Th1 response; the result is an imbalance between Th1 and Th2 activation. PMID- 15345200 TI - Sick building syndrome in like symptoms in emergency prefabricated accommodation. AB - The present study investigated the sources of discomfort and the symptoms reported by earthquake victims residing in temporary emergency prefabricated accommodation (prefab). The investigation was carried out by means of a questionnaire. 203 prefab occupants and 13 inhabitants of houses, who were chosen as reference population, replied in winter and 233 prefab occupants and 154 inhabitants of houses replied in summer. In both seasons more people living in prefabs indentified dry air, stuffy air, stale air, dust, dampness, uncomfortable temperature and bad odours as sources of discomfort. They also complained of general symptoms (headache, irritablility, insomnia, difficulty in concentration) and irritative symptoms of the eyes, upper and lower airways and skin. Multiple regression analysis identified the type of accommodation as the variable that most influenced the onset of general, ocular, upper and lower airway symptoms. Intrinsic characteristics of the prefabs (being constructed with synthetic materials, combustion sources, poor ventilation and insulation) and psychosocial factors e. losing their home, could have contributed to the onset of symptoms. PMID- 15345201 TI - Naive (CD4+CD45RA+) T cell subpopulation is susceptible to various types of hazardous substances in the workplace. AB - To find the most susceptible lymphocyte subpopulation with exposure to various occupational hazardous substances, we reanalyzed our previous data. We measured T, B and natural killer NK cell subpopulations by means of flowcytometry among workers who were exposed to aromatic amines, chromate, mercury lead and organic solvents. The exposed/non-exposed ratio was defined as the mean value of the absolute number of lymphocyte subpopulations in the exposed workers divided by the corresponding value of the non-exposed control group in 5 studies reported previously by us. The ratio of CD4+CD45RA+ T cells in all the exposed groups examined in this study was less than 1.0, and the mean of these ratio was 0.77, which was the lowest among lymphocyte subpopulations examined. The mean ratio was 1.06 for CD4+CD29+ T cells, 0.91 for CD4+ T cells, 0.99 for CD8+ T cells, 0.93 for CD3+ T cells, 0.99 for CD19+B cells, 0.96 for CD57-CD16+ NK cells, 0.82 for CD57+CD16+ NK cells, 1.25 for CD57+CD16- NK cells and 0.89 for total lymphocytes. With regard to the difference between exposed and non-exposed workers the estimated overall difference in the mean absolute number of CD4+CD45RA+ T cell subpopulation between exposed and non-exposed workers among five studies was -135 ((% confidence interval, CI: (-194,-75)) cells/mm3 (p<0.01). In conclusion, naive (CD4+CD45RA+) T cell subpopulation was the most susceptible to the effects of various toxic substances as compared with other lymphocyte subpopulations. This lympohocyte subpopulation may be useful for monitoring immune system of workers exposed to various types of substances in the workplace. PMID- 15345202 TI - "In vitro" comparative immune effects of different titanium compounds. AB - Exposure to Ti compounds is today an occupational and environmental health hazard. Object of this study was to determine "in vitro" effects of different Ti salts on cultured human peripheral blood mononuclear cells (PBMC) proliferation and cytokine release. 10(-4) and 10(-7) M Ti compounds did not modify spontaneous PBMC proliferation. Ti dioxide (a biocompatible material and sunscreen component) did not exert effects on phytoemagglutinin (PHA) stimulated PBMC proliferation and on PHA stimulated IFN-gamma and TNF-alpha release from PBMC. On the other hand, 10(-4) M Ti oxalate (with wide industrial applications) and Ti ascorbate (used mainly in agriculture) inhibited about 70% the PHA stimulate PBMC proliferation; both these Ti compounds at 10(-4) and 10(-7) M concentrations significantly inhibited TNF-alpha release, while only Ti oxalate inhibited that of IFN-gamma. Titanocene (used in chemotherapy) did no exert effects on PBMC proliferation but markedly inhibited IFN-gamma and TNF-alpha release. On the whole, this study demonstrates that Ti dioxide is not immunotoxic; Ti oxalate shows marked immunotoxicity; titanocene exerts selective toxicity on cytokine release but not on PBMC proliferation, while Ti ascorbate affects TNF-alpha release from PBMC but not iFN-gamma release. In conclusion, the data show that immunotoxicity fo Ti depends on speciation. PMID- 15345203 TI - Reversibility of thymulin production impairment by L-arginine supplementation in mice exposed to inorganic mercury. AB - Immunotoxicological effects fo mercury on peripheral immune system are known. We had previously in vitro found that mercuric chloride inhibits thymulin production in mouse thymus cultures at concentrations as low as 10(-8) M. In this study, thymus efficiency, assessed as production of active and total thymulin, was evaluated in vivo using young mice that were injected sc every 3 days for 4 weeks with saline containing mercuric chloride at different concentrations (0 -controls , 0.001 or 1.0 mg HgCl2/kg body weight). The results show that both the doses are able to cause a significant reduction in active and total thymulin production. Since arginine enhances immune efficiency some of the animals also received a diet supplemented with arginine in order to evaluate a possible role of arginine during mercury intoxication. The data show that arginine has a protective effect on thymic endocrine efficiency. Mice, treated with the lowest dose of mercury and receiving and arginine supplemented diet, produced active and total thymulin like mercury untreated mice. Arginine is an aminoacid which may be found in various amounts in different foods, some foods are particularly rich in arginine i.e. peanuts, stock fish. We suggest that the daily arginine intake may account for individual susceptibility to the mercury-induced immunological effects which are found in mercury occupationally exposed workers. PMID- 15345204 TI - Pancreatic Transplantation: Beta Cell Replacement. AB - Diabetes is a leading cause of morbidity and mortality worldwide. Complications of diabetes including renal failure, retinopathy, neuropathy, and cardiovascular disease limit both survival and quality of life. Pancreatic transplantation can restore euglycemia thereby stabilizing or even reversing secondary complications of diabetes as well as improving quality of life particularly in patients with labile diabetes. Recent evidence also shows an improved survival in diabetic patients that undergo pancreatic transplantation when combined with a kidney transplant. Pancreatic transplantation should more properly be referred to as beta cell replacement as the field today encompasses both whole organ and islet cell transplantation. We have outlined herein the indications and contraindications to islet or whole organ pancreas transplantation and we have described periprocedure care and short- and long-term prognosis. PMID- 15345205 TI - Advances in the Genetic Screening, Work-up, and Treatment of Pancreatic Cancer. AB - Familiarity with the updated results in genetic screening and work-up presented here is essential to early diagnosis and possible cure. In the metastatic setting, we most frequently begin with the GTX regimen, consisting of Gemcitabine, Taxotere, and Xeloda. The regimen is based on our laboratory data demonstrating a synergistic increase in cell killing of pancreatic cancer cell lines. The combination takes advantage of the selective cell cycle effects of each of the three drugs. In our initial experience, we have seen a response rate of 40% at metastatic sites and 31% at the primary site after nine cycles of GTX. We are now conducting a formal phase II protocol to confirm these results. The median survival of this group of patients (at least 10.4 months) is as long as, or longer than other currently used regimens. In those patients who do not tolerate GTX or progress despite the regimen, we have found that a regimen of the same three drugs, administered on a different schedule, can produce responses. In the neoadjuvant (unresectable) setting, we treat with GTX initially and then follow with radiation; gemcitabine is used as a radiosensitizer during this treatment. An aggressive surgical approach with a team of surgeons were able to resect for cure 12 of the 16 patients who were initially unresectable; one year survival of these 12 was 100%; 2 year survival was 50%. Future work in this disease should focus on targeted agents such as bevacizumab. PMID- 15345206 TI - Neuroendocrine Tumors of the Pancreas. AB - Neuroendocrine tumors (NETs) are rare tumors of the endocrine pancreas that require a high degree of suspicion for timely diagnosis. Diagnosis is often delayed due to the nonspecific and intermittent presentation of symptoms. As many as 45% to 55% of tumors are nonfunctional and are typically diagnosed secondary to mass effect related symptoms or found incidentally. Functional tumors often are symptom specific and are diagnosed at an earlier stage than nonfunctional tumors. The challenging aspects of treating NETs are localizing the tumors, treating extensive or metastatic disease, and palliating symptoms. Most NETs have an indolent course, and aggressive multimodality treatment is often indicated and encouraged. PMID- 15345207 TI - Treatment of Post-pancreaticoduodenectomy Complications. AB - Pancreaticoduodenectomy is a complex operation that is becoming more common as treatment for both malignant and benign diseases. While postoperative mortality has improved over the last two decades, morbidity continues to remain high. The improvement in mortality is attributed to better perioperative care, including aggressive early diagnosis of complications and a multidisciplinary approach to their treatment. In addition to a high clinical suspicion for postoperative complications, ready access to state of the art diagnostic radiology and endoscopy are essential for the early and accurate diagnosis of complications. After the diagnosis of a complication is made, the patient should have expertise available in interventional radiology, gastroenterology, and hepatobiliary and pancreatic surgery. Optimal treatment may involve any one of these specialties or an orchestrated effort from them all. The need for a critical mass of expertise in many specialties and subspecialties has prompted the development of new tertiary centers devoted to hepaticopancreaticobiliary (HPB) diseases. These HPB centers are becoming more prevalent in the United States as the value of this subspecialty is becoming more recognized. These HPB centers should continue to show improvements in quality and cost of care in treating complex liver, pancreatic, and biliary diseases that have rapidly evolving treatment options. PMID- 15345208 TI - Treatment Options for Hepatitis C Infection in Children. AB - Multiple factors support treatment of hepatitis C virus (HCV) infection in children. These factors include the anticipated long duration of infection after early acquisition, relatively good tolerance of antiviral medications, and avoidance of social stigmatization. Nevertheless, careful selection of appropriate candidates for therapy is important. If a contraindication to current therapeutic agents is present, treatment should be withheld until this has resolved or until new agents are available. Children without contraindications to the medications used for HCV should undergo liver biopsy to determine the presence and degree of fibrosis. In the absence of fibrosis, treatment may be deferred. If any degree of hepatic fibrosis is present, antiviral therapy for HCV should be considered. At present, in the United States, the only therapy approved for children by the Food and Drug Administration (FDA) is a combination of interferon (IFN) alfa-2b and ribavirin. No safe therapies have been established for children younger than 3 years of age. Pegylated interferon in combination with ribavirin may be considered in adolescents older than 16 years of age who are post-pubertal, or in younger children in the context of clinical trials. Multicenter trials are currently underway to determine the safety and effectiveness of other forms of therapy for HCV infection in children. PMID- 15345209 TI - Management of Fecal Incontinence in Children Without Functional Fecal Retention. AB - The management of the fecal incontinence in children is difficult, and its social consequences are usually devastating. The general objectives of any bowel program are to produce social continence, predictability, and eventually independence. How to achieve those goals depends in part on the underlying condition. In children, fecal incontinence can occur from a variety of conditions. The most common is overflow incontinence from functional fecal retention, but it can also occur in otherwise healthy children with functional nonretentive fecal soiling or in children with organic causes of fecal incontinence, such as congenital malformations, or any other condition affecting the anorectum, anal sphincters, or the spinal cord. The therapeutic regimen that is recommended in patients with nonretentive fecal soiling consists of explanation and support for the child and parents, a nonaccusatory approach, and a toilet training program with a rewarding system. Biofeedback does not play an important role, and laxatives need to be used with caution, as they may exacerbate the incontinence. For those patients with congenital/neuropathic incontinence a combination of maneuvers to change stool consistency, colonic transit, anorectal function, and rectosigmoid evacuation is used. Stool consistency can be changed with the use of dietary interventions or medications. Stool transit can be slowed (antimotility agents) or accelerated (laxatives) with the use of medications. Anorectal function can be improved with the use of biofeedback or procedures to alter sphincter pressure, and the production of a bowel movement can be induced with maneuvers to empty the sigmoid (suppositories, enemas). With the recent advent of the Antegrade Colonic Enema (ACE), the patient is then able to be predictable and independent. This procedure creates a continent conduit from the skin to the cecum that can be catheterized or accessed for self-administration of enemas. The ACE has revolutionized the treatment of children with fecal incontinence. PMID- 15345210 TI - Targeting Therapy in Pediatric Inflammatory Bowel Disease. AB - Progress in our understanding of the interaction between the environment and the immune system in disease pathogenesis has led to major advancements in the area of inflammatory bowel disease (IBD) therapeutics. Biotechnology is keeping pace with these scientific advances. Current therapies target the various elements of the inflammatory cascade implicated in the pathogenesis of IBD. The anti inflammatory and immunomodulatory properties of the pharmacologic therapies used in IBD vary from actions that are extremely broad to those that are cellular or cytokine specific. Despite the various therapeutic options available for IBD patients, chosen therapies should be based on the overall treatment goal for individual patients. Therapeutics can be broadly categorized as induction therapies (goal to treat active disease) and maintenance therapies (goal to prevent relapse of disease). The modern thinking behind drug development is that IBD therapy should be disease modifying so to avoid complications and alter the long term natural history of disease. This review will cover both current and emerging agents and highlight the pathogenesis of IBD and how it relates to therapeutic targets. PMID- 15345211 TI - Treatment of Helicobacter pylori in Pediatrics. AB - Helicobacter pylori (H. pylori) is among the most common bacterial infections in humans. In 1982, H. pylori was discovered by Marshal and Warren, demonstrating an association between H. pylori and ulcer disease. H. pylori is a gram-negative, S shaped rod that produces enzymes like urease, catalase and oxidase. The mechanism of acquisition and transmission of H. pylori is unclear, although the most likely mode of transmission is fecal-oral and oral-oral. The mode of transmission is supported by studies that demonstrate viable H. pylori organisms can be cultured from the stool or vomitus of infected patients. Risk factors such as minimal education and low socio-economic status during childhood affect the prevalence. Children infected with H. pylori develop histologic chronic active gastritis despite the fact that they are generally asymptomatic. A small percentage of these children will go on to develop peptic ulcer disease, and even gastric cancer. In contrast, the association of abdominal pain and H. pylori infection remains controversial. In the year 2000, the North American Society of Pediatric Gastroenterology guidelines on H. pylori reported that there is no evidence demonstrating a link between H. pylori-associated gastritis and abdominal pain, except in rare cases in which gastric or duodenal ulcer disease is present. Currently, treatment with a combination of two antimicrobial agents in conjunction with a proton pump inhibitor (PPI) continues to be recommended for the treatment of H. pylori associated peptic ulcer disease. PMID- 15345212 TI - Treatment of Autoimmune Hepatitis: Current and Future Therapies. AB - Autoimmune hepatitis is an uncommon liver disease affecting children and adults. Early diagnosis and start of treatment improve the response and long-term outcome. Initial treatment depends on patient's age and the clinical, laboratory, and histological features that allow the prediction of the response, as well as the presence or absence of associated extrahepatic disorders. In specialized centers, short-term cyclosporine is used safely and successfully to control the liver inflammatory process. Low doses of prednisone in association with azathioprine are sufficient to sustain the response. Maintenance treatment must be administered for several years, and withdrawal can be attempted after at least 4 years of a complete and sustained response. Future research should focus on the recovery of immune homeostasis in these patients by less aggressive means. PMID- 15345213 TI - Mysterious origin of plasmacytoid dendritic cell precursors. AB - The origin of plasmacytoid dendritic cell precursors (pDC) has been controversial. This manuscript shows that CLP and CMP both can give rise to pDCs. pDCs derived from CMP express RAG gene products and show IgHD-J rearrangement, suggesting that pDC represent a unique lineage whose gene expression program shows substantial plasticity. PMID- 15345214 TI - Bidirectional promoters regulate the monoallelically expressed Ly49 NK receptors. AB - Members of the Ly49 gene family of natural killer (NK) cell receptors in mice are expressed in seemingly stochastic combinations such that each NK cell expresses a handful of family members. A transcriptional switch appears to establish this interesting pattern of expression. PMID- 15345215 TI - Cutaneous gammadelta T cells-selecting a destination and asking for directions in the thymus. AB - T cell receptor-defined subsets of gammadelta T cells differentially localize in cutaneous, intestinal, or reproductive epithelium. In this issue of Immunity, Xiong et al. show that murine Vgamma3+ T cells undergo TCR-dependent selection in the fetal thymus that drives expression of a cutaneous homing receptor. PMID- 15345216 TI - The Cbl family and other ubiquitin ligases: destructive forces in control of antigen receptor signaling. AB - Regulation of tyrosine kinase-mediated cellular activation through antigen receptors is of great biological and practical significance. The evolutionarily conserved Cbl family ubiquitin ligases have emerged as key negative regulators of activated tyrosine kinase-coupled receptors, and their impaired function switches a normal immune response into autoimmunity. Cbl proteins facilitate the ubiquitinylation of activated tyrosine kinases and other signaling proteins and of the signaling chains of receptors themselves; monoubiquitin tag promotes sorting of activated receptors and associated proteins into internal vesicles of the multivesicular body, facilitating their lysosomal degradation, whereas polyubiquitin tag promotes proteasomal degradation. Notably, increased expression of Cbl proteins and other ubiquitin ligases is a component of anergic signaling program in T cells. Thus, controlled destruction of the signaling apparatus has emerged as a key to fine-tuning antigen receptor signaling. Further studies of this pathway are likely to elucidate the pathogenesis of autoimmune diseases and offer new therapeutic targets. PMID- 15345217 TI - The mitogen-induced increase in T cell size involves PKC and NFAT activation of Rel/NF-kappaB-dependent c-myc expression. AB - Cell growth during the G1 stage of the cell cycle is partly controlled by inducing c-myc expression, which in B cells is regulated by the NF-kappaB1 and c Rel transcription factors. Here, we show that c-myc-dependent growth during T cell activation requires c-Rel and RelA and that blocking this growth by inhibiting protein kinase C theta (PKCtheta) coincides with a failure to upregulate c-myc due to impaired RelA nuclear import and inhibition of NFAT dependent c-rel transcription. These results demonstrate that different Rel/NF kappaB dimers regulate the mitogenic growth of mature T and B cells, with a signaling pathway incorporating PKCtheta and NFAT controlling c-Rel/RelA-induced c-myc expression in activated T cells. PMID- 15345218 TI - Endothelial and hematopoietic cell fate of human embryonic stem cells originates from primitive endothelium with hemangioblastic properties. AB - The cellular organization and relationships among precursors that initiate embryonic angiogenesis and hematopoiesis in the human have yet to be characterized. Here, we identify a subpopulation of primitive endothelial-like cells derived from human embryonic stem cells (hESCs) that express PECAM-1, Flk 1, and VE-cadherin, but not CD45 (CD45negPFV cells), and that are uniquely responsible for endothelial and hematopoietic development. Molecular profiling of CD45negPFV cells is consistent with endothelial and hematopoietic competency. Clonal isolation demonstrates that the CD45negPFV population includes bipotent cells with endothelial and hematopoietic capacity. We suggest that human hematopoiesis and endothelial maturation originate exclusively from a subset of embryonic endothelium that possesses hemangioblastic properties and offers a model system to study these lineage relationships in the human. PMID- 15345219 TI - Plasmacytoid dendritic cells activate lymphoid-specific genetic programs irrespective of their cellular origin. AB - The developmental origin of type I interferon (IFN)-producing plasmacytoid dendritic cells (PDCs) is controversial. In particular, the rearrangement of immunoglobulin heavy chain (IgH) genes in murine PDCs and the expression of pre-T cell receptor alpha (pTalpha) gene by human PDCs were proposed as evidence for their "lymphoid" origin. Here we demonstrate that PDCs capable of IFN production develop efficiently from both myeloid- and lymphoid-committed progenitors. Rearranged IgH genes as well as RAG transcripts were found in both myeloid- and lymphoid-derived PDCs. The human pTalpha transgenic reporter was activated in both myeloid- and lymphoid-derived PDCs at a level comparable to pre-T cells. PDCs were the only cell population that activated murine RAG1 knockin and human pTalpha transgenic reporters outside the lymphoid lineage. These results highlight a unique developmental program of PDCs that distinguishes them from other cell types including conventional dendritic cells. PMID- 15345220 TI - Identification of probabilistic transcriptional switches in the Ly49 gene cluster: a eukaryotic mechanism for selective gene activation. AB - Murine natural killer cells selectively express members of the Ly49 family of class I MHC receptors; however, the molecular mechanism controlling probabilistic expression of Ly49 proteins has not been defined. A pair of overlapping, divergent promoters discovered in the Ly49g gene functions as a molecular switch that can produce a forward transcript containing the coding region of the gene (on position) or a noncoding transcript in the opposite direction (off position), and this element maintains transcription in the chosen direction. Competition of C/EBP and TBP transcription factors for overlapping binding sites determines the relative strength of the competing promoters and the probability of transcription in a given direction. Similar elements precede all Ly49 family members, and the relative strength of the forward promoter in each inhibitory Ly49 gene correlates with the percentage of natural killer cells that express a given receptor, supporting a promoter competition model of selective gene activation. PMID- 15345221 TI - Signaling through Itk promotes T helper 2 differentiation via negative regulation of T-bet. AB - The Tec family tyrosine kinase, Itk, is critical for PLC-gamma1 activation downstream of the TCR. Studies of Itk-/- mice have demonstrated a requirement for Itk in Th2 cytokine production and protective immunity to parasitic infections. Here we address the mechanism by which Itk regulates Th2 differentiation. We find that naive Itk-/- CD4+ T cells respond normally to cytokine skewing signals and can differentiate efficiently into either Th1 or Th2 lineage cells. In the absence of skewing cytokines, wild-type CD4+ T cells stimulated with low-avidity ligands preferentially express GATA-3 mRNA and differentiate into Th2 cells. Under these same stimulation conditions, Itk-/- T cells produce large amounts of T-bet mRNA and differentiate into IFN-gamma-producing cells. Furthermore, Itk is upregulated during Th2 differentiation, while Rlk, a related Tec kinase, disappears rapidly from differentiating Th2 cells. Together, these findings provide a molecular explanation for the essential role of Itk in Th2 differentiation. PMID- 15345222 TI - XBP1, downstream of Blimp-1, expands the secretory apparatus and other organelles, and increases protein synthesis in plasma cell differentiation. AB - The differentiation of B cells into immunoglobulin-secreting plasma cells is controlled by two transcription factors, Blimp-1 and XBP1. By gene expression profiling, we defined a set of genes whose induction during mouse plasmacytic differentiation is dependent on Blimp-1 and/or XBP1. Blimp-1-deficient B cells failed to upregulate most plasma cell-specific genes, including xbp1. Differentiating xbp1-deficient B cells induced Blimp-1 normally but failed to upregulate genes encoding many secretory pathway components. Conversely, ectopic expression of XBP1 induced a wide spectrum of secretory pathway genes and physically expanded the endoplasmic reticulum. In addition, XBP1 increased cell size, lysosome content, mitochondrial mass and function, ribosome numbers, and total protein synthesis. Thus, XBP1 coordinates diverse changes in cellular structure and function resulting in the characteristic phenotype of professional secretory cells. PMID- 15345223 TI - Activation of PPARgamma specifies a dendritic cell subtype capable of enhanced induction of iNKT cell expansion. AB - Little is known of the transcriptional events controlling the differentiation and function of dendritic cells (DC). We found that the ligand-activated transcription factor Peroxisome Proliferator Activated Receptor gamma (PPARgamma) is immediately upregulated after the induction of monocyte-derived DC differentiation. Activation of PPARgamma changed the expression pattern of cell surface receptors and enhanced the internalizing activity of DC. Unexpectedly, we found that CD1 glycoproteins, a class of molecules responsible for the presentation of self and foreign modified lipids, were coordinately regulated by PPARgamma activation. CD1a levels were reduced, while CD1d expression was induced. Enhanced expression of CD1d was coupled to the selective induction of invariant natural-killer T cell (iNKT cell) proliferation in the presence of alpha-GalCer. These results suggest that PPARgamma orchestrates a transcriptional response leading to the development of a DC subtype with increased internalizing capacity, efficient lipid presentation, and the augmented potential to activate iNKT cells. PMID- 15345224 TI - TLR9-dependent recognition of MCMV by IPC and DC generates coordinated cytokine responses that activate antiviral NK cell function. AB - Natural interferon-producing cells (IPC) respond to viruses by secreting type I interferon (IFN) and interleukin-12 (IL-12). Toll-like receptor (TLR) 9 mediates IPC recognition of some of these viruses in vitro. However, whether TLR9-induced activation of IPC is necessary for an effective antiviral response in vivo is not clear. Here, we demonstrate that IPC and dendritic cells (DC) recognize murine cytomegalovirus (MCMV) through TLR9. TLR9-mediated cytokine secretion promotes viral clearance by NK cells that express the MCMV-specific receptor Ly49H. Although depletion of IPC leads to a drastic reduction of the IFN-alpha response, this allows other cell types to secrete IL-12, ensuring normal IFN-gamma and NK cell responses to MCMV. We conclude that the TLR9/MyD88 pathway mediates antiviral cytokine responses by IPC, DC, and possibly other cell types, which are coordinated to promote effective NK cell function and MCMV clearance. PMID- 15345225 TI - Positive selection of dendritic epidermal gammadelta T cell precursors in the fetal thymus determines expression of skin-homing receptors. AB - The role of cellular selection in the development of gammadelta T cells remains unclear. Knockout mice lacking a subset of Vgamma genes, including Vgamma3, contain abundant gammadelta T cells but are devoid of dendritic epidermal gammadeltaT cells (DETCs), which normally express an invariant Vgamma3/Vdelta1 gammadelta TCR. A rearranged Vgamma2 transgene restored DETC development, but the restored DETCs selectively expressed a unique Vdelta gene other than Vdelta1, indicating that DETC development involves TCR-based selection. In both normal and transgenic/knockout mice, specific DETC precursors in the fetal thymus were activated and expressed the IL-15 receptor beta chain, skin-homing receptors, and thymic exiting receptors. In vitro activation of irrelevant precursors also led to upregulation of the skin-homing receptor, providing an explanation for how thymic selection is coordinated with development of epidermal gammadelta T cells. PMID- 15345226 TI - Involvement of Mycobacterium smegmatis undecaprenyl phosphokinase in biofilm and smegma formation. AB - We describe a Mycobacterium smegmatis mutant with impaired biofilm and smegma formation. A gene homologous to Escherichia coli bacA, which has been proposed to play a role as undecaprenyl phosphokinase (Upk) was unmarked in-frame deleted from M. smegmatis. Though Upk is involved in cell wall synthesis, the surface of the mutant strain appeared virtually comparable to that of the wild type by electron microscopy. The absence of Upk influenced colony morphology and bacitracin resistance. The M. smegmatis Deltaupk mutant developed a biofilm characterized by scattered islands of bacteria distinct from the completely covered biofilm surface observed for wild-type bacteria. We further demonstrate biological consequences of upk deletion for smegma development in an in vivo model. These results suggest the upk gene to be essential in biofilm and smegma development. PMID- 15345227 TI - Bacteriological follow-up of pulmonary tuberculosis treatment: a study with a simple colorimetric assay. AB - The viability of Mycobacterium tuberculosis (MTB) in serial sputum specimens from persistently smear positive patients was evaluated. The assay was based on oxidation-reduction of Alamar Blue and Malachite Green dyes that change their color in response to MTB growth. A total of 280 sputum specimens from 40 persistently smear positive TB patients and 40 sputa from non-tuberculosis patients were digested, decontaminated and examined microscopically. To check the MTB viability, the sediments from decontaminated samples were inoculated into three culture media: Lowenstein-Jensen (LJ) slants, Alamar Blue and Malachite Green culture tubes. We found that out of 280 smear positive specimens, the LJ culture was positive in 124 (44%). The numbers of correctly identified S+/C+ cases by Alamar Blue and Malachite Green were 118 (95%) and 116 (93%), respectively. The mean time required for reporting the positive signal in Alamar Blue culture tubes was 9 versus 11 days by Malachite Green culture tubes. In the standard LJ culture media the average detection time was 27 days (P < 0.05). The sensitivity of LJ was 99%, Alamar Blue 95% and Malachite Green 93%. The specificity was 100%, 92% and 93%, respectively. The oxidation-reduction method is rapid, sensitive and inexpensive in monitoring the treatment response of patients with pulmonary TB. Thus, using this method can be of paramount importance, particularly in resource-constrained areas. PMID- 15345228 TI - Leishmania braziliensis isolates differing at the genome level display distinctive features in BALB/c mice. AB - Leishmania braziliensis is the species responsible for the majority of cases of human cutaneous leishmaniasis in Brazil. In the present study, L. braziliensis isolates from two different geographic areas in Brazil were studied by RAPD, using arbitrary primers. We also evaluated other biological features of these two isolates. We compared (a) the clinical features they initiate or not once delivered subcutaneously as stationary-phase promastigotes in the footpad of BALB/c mice; (b) the parasite load in both the footpad and the draining lymph node; (c) the cytokines present in the supernatant of cultures of the cell suspensions from the draining lymph nodes; and (d) the cell types present at the site of parasite delivery. The results show that the L. braziliensis strain from Ceara (H3227) is genotypically different from the L. braziliensis strain from Bahia (BA788). H3227-parasitized mice developed detectable lesions, whereas BA788 parasitized mice did not. Fifteen days post parasite inoculation there was an increase in the numbers of macrophages and lymphocytes in the footpads, whatever the parasite inoculum. Parasite load at the inoculation site--namely the footpad- did not differ significantly; in draining lymph nodes, however, it increased over the period under study. Early after parasite inoculation, the cells recovered from the draining lymph nodes of BA788-parasitized mice produced higher levels of IFN-gamma, a feature coupled to a higher number of NK cells. Later, after the parasite inoculation, there was an increased content of IL-12p70 and IL-10 in the supernatant of cells recovered from the lymph nodes of H3227-parasitized mice. This comparative analysis points out that L. braziliensis isolates differing in their genomic profiles do establish different parasitic processes in BALB/c mice. PMID- 15345229 TI - Human dendritic cells are less potent at killing Candida albicans than both monocytes and macrophages. AB - Dendritic cells (DC) function as professional phagocytes to kill Candida albicans and subsequently present it to the adaptive immune system. Monocytes, macrophages and DC were generated from five individual donors and their Candida-killing capacity and cytokine release were assessed. Compared to monocytes and macrophages, DC from healthy volunteers were significantly less effective in C. albicans--stimulated cytokine release, killing of C. albicans blastoconidia and damaging of C. albicans hyphae. In conclusion, while important as antigen presenting cells and initiators of the adaptive immune system, DC are poor in both intracellular killing and damaging of C. albicans hyphae. Effective handling of large numbers of C. albicans is the prime task of the innate immune system consisting of large numbers of neutrophils and monocytes. PMID- 15345230 TI - Lipopolysaccharide needs soluble CD14 to interact with TLR4 in human monocytes depleted of membrane CD14. AB - Toll-like receptors recognize specific patterns of microbial components and regulate the activation of both innate and adaptive immunity. TLR4 recognizes lipopolysaccharide (LPS) in monocytes/macrophages with the help of other molecules like CD14 and MD-2, which indicates that the functional LPS receptor forms a large complex. The functional relationship between the components has been the subject of debate, as have the modifications induced by the ligand in the expression of some of these components. Moreover, as for other members of this family of receptors, the possible direct interaction of receptors and their ligands is a matter of discussion. In this paper we address the question of whether the expression of some of the components influences the expression of the rest. Human monocytes in which CD14 has been downregulated through interference in the turnover of the molecule at the Golgi level, show normal membrane TLR4 expression, when compared with control cells. On the other hand, LPS alters membrane TLR4 expression by monocytes devoid of membrane CD14 only in the presence of human serum. The effect of serum is blocked by anti-CD14 monoclonal antibodies, which strongly suggests a functional role for soluble CD14/LPS complexes in the interaction with TLR4. Our data add information on the relationship between the components of the LPS receptor and the characteristics of the interaction of LPS and TLR4 in cells devoid of membrane CD14. PMID- 15345231 TI - Natural and experimental infection of immunocompromised rhesus macaques (Macaca mulatta) with the microsporidian Enterocytozoon bieneusi genotype D. AB - Microsporidia are obligate intracellular parasites that cause opportunistic infections in AIDS and other immunocompromised patients. Eight simian immunodeficiency virus (SIV)-infected rhesus macaque monkeys (Macaca mulatta) were inoculated orally with Enterocytozoon bieneusi spores isolated from intestinal lavage fluid of an AIDS patient (genotype D) to study the natural history of this infection. Four monkeys were already naturally infected with E. bieneusi (also genotype D), and were included to determine if a second inoculum affected the course of illness. Spore shedding was detected in feces of all eight monkeys within the first week of experimental infection. Five monkeys died within 3.5 months of experimental E. bieneusi inoculation. Three of these five monkeys began the study with CD4+CD29+ T cell levels well below 20% of total T lymphocytes. Deaths were due to a variety of AIDS-related manifestations. Microsporidia did not appear to directly contribute to mortality but may have contributed to morbidity. At necropsy, microsporidia were found in bile and tissue sections of the gallbladder but not in the gut, kidneys, or liver. The percent CD4+CD29+ levels of the last three monkeys remained near the level observed at the time of inoculation. These monkeys lived more than 2 years after the end of the study and continued to shed spores. This study corroborates previous reports that E. bieneusi can be reliably transmitted to SIV-infected rhesus monkeys but indicates that the use of SIV-infected monkeys for the study of microsporidiosis is complicated by the confounding effect of other opportunistic or AIDS-related infections. PMID- 15345233 TI - Nosocomial lung infections in adult intensive care units. AB - Nosocomial respiratory tract infections are the leading type of nosocomial infections. Despite the development of new antibiotic therapies, they are associated with an increased morbidity and mortality. Patients with comorbidities are especially predisposed to acquire these infections, as are patients exposed to respiratory therapy. Aspiration of colonized secretions from the oropharynx is the main mechanism of infection development. Barrier techniques to reduce aspiration and antimicrobial agents to alter bacterial flora are important in preventing pneumonia episodes. The initial institution of an adequate antibiotic regimen is a determinant of outcome. Nosocomial pneumonias are often difficult to treat due to antibiotic-resistant bacteria. Antibiotic policies are crucial in avoiding a progression in antibiotic resistance. PMID- 15345234 TI - Intra-abdominal infections. AB - Intra-abdominal infections in the hospitalized patient differ from those arising in the community in their clinical presentation, sites of involvement, and characteristic microbiology. They are also associated with greater morbidity and mortality. New onset organ dysfunction, more than acute abdominal pain and tenderness, is the predominant clinical manifestation. Successful management depends on aggressive resuscitation and hemodynamic support, administration of adequate antimicrobial therapy, and the timely use of source control measures appropriate to the clinical situation. PMID- 15345235 TI - Catheter-associated urinary tract infections in intensive care units. AB - The purpose of this review is to analyze literature concerning the diagnosis, prevention, and management of catheter-associated urinary tract infection (CAUTI) occurring in patients hospitalized in the intensive care unit (ICU). Analysis was performed from personal and "Pubmed" data, crossing the following keywords: "urinary tract infection", "catheter', and "intensive care unit". Few clinical trials including ICU patients were found despite the abundance of expert opinions. There is no consensus on the use of urinary reagent tests for diagnosis. The prevention of CAUTI in ICU patients does not require expensive devices. Neither complex closed drainage systems nor silver-coated urinary catheters have demonstrated efficacy in comparative randomized clinical trials. Bladder irrigation should not be used, except when an obstruction of the catheter is highly likely. The administration of prophylactic antimicrobial therapy, although effective in reducing the incidence of urinary bacteria, cannot be recommended in ICU patients. The management of CAUTI in ICU patients has not been evaluated in clinical trials. The level of evidence provided in this field is weak, and underlines the need for randomized studies to improve management of patients. PMID- 15345236 TI - Catheter-related infections. AB - Nosocomial infections are a leading cause of morbidity and mortality among hospitalized patients. These infections have made newspaper headlines recently in many countries, and both patients and their relatives are now perfectly aware of their existence and of the risks which are inherent to any medical activity. However, significant improvements in the knowledge of the pathophysiology and epidemiology of nosocomial infections allow us to prevent them efficiently. Accordingly, they should no longer be considered as an inevitable tribute to pay to the continuous progress of medicine, but as a real challenge in the process of improving the quality of patient care. This is particularly the case for bloodstream infections, of which at least 80% are considered to be catheter associated. This paper reviews the epidemiology and impact of infections associated with the use of intravenous catheters. Principles of therapy are reviewed, as well as major aspects of prevention. PMID- 15345237 TI - Microbiology of nosocomial infections: progress and challenges. AB - Technological advances have expanded the role of the clinical microbiology laboratory in supporting management and control of health-care-associated infections (HAI). Diagnostic capabilities have been complemented by new methods for characterisation of microbial antigens and nucleic acids. Genomic "fingerprinting" enables the study of transmission of nosocomial pathogens. Monitoring of their antimicrobial resistance patterns has gained critical importance. However, cost-effective application of microbiological testing for improved treatment and prevention of HAI requires further study. PMID- 15345238 TI - RhoA, Rac1, and cdc42 intracellular distribution shift during hippocampal neuron development. AB - Differences in cytoskeleton organization are key determinants of the architecture and dynamics of axons and dendrites. This is most clearly illustrated by the diverse pools of microtubule-associated proteins in axons and dendrites. Whether similar polarized organization occurs for actin regulatory proteins remains to be determined. To address this issue, we analyzed the intracellular distribution of the Rho GTPases, RhoA, Rac1, and cdc42 in hippocampal neurons in culture. We report that all three Rho members are evenly distributed during the time of axon and dendrite sprouting. This is not the case in mature neurons, as RhoA enriches in dendrites, Rac1 in axons, and Cdc42 is equally abundant in both domains. Polarized segregation of the actin regulatory machinery in mature neurons might play an important role in axonal and dendritic architectural plasticity. PMID- 15345239 TI - p59(fyn)-mediated phosphorylation regulates the activity of the tissue-specific splicing factor rSLM-1. AB - The Sam68-like mammalian protein SLM-1 is a member of the STAR protein family and is related to SAM68 and SLM-2. Here, we demonstrate that rSLM-1 interacts with itself, scaffold-attachment factor B, YT521-B, SAM68, rSLM-2, SRp30c, and hnRNP G. rSLM-1 regulates splice site selection in vivo via a purine-rich enhancer. In contrast to the widely expressed SAM68 and rSLM-2 proteins, rSLM-1 is found primarily in brain and, to a much smaller degree, in testis. In the brain, rSLM-1 and rSLM-2 are predominantly expressed in different neurons. In the hippocampal formation, rSLM-1 is present only in the dentate gyrus, whereas rSLM-2 is found in the pyramidal cells of the CA1, CA3, and CA4 regions. rSLM-1, but not rSLM-2, is phosphorylated by p59(fyn). p59(fyn)-mediated phosphorylation abolishes the ability of rSLM-1 to regulate splice site selection, but has no effect on rSLM-2 activity. This suggests that rSLM-1-positive cells could respond with a change of their splicing pattern to p59(fyn) activation, whereas rSLM-2-positive cells would not be affected. Together, our data indicate that rSLM-1 is a tissue specific splicing factor whose activity is regulated by tyrosine phosphorylation signals emanating from p59(fyn). PMID- 15345240 TI - Specific beta1 integrins mediate adhesion, migration, and differentiation of neural progenitors derived from the embryonic striatum. AB - Early inductive signals within the embryonic mammalian forebrain establish two major germinal regions along the dorsal-ventral axis. The dorsal germinal zone eventually forms the cerebral cortex while the ventral ganglionic eminence primarily forms the striatum and globus pallidus. The mechanisms leading to patterning of specific forebrain structures from these distinct germinal regions are not fully understood but may involve the adhesive and migratory properties of regionally specified cells and their interactions with the extracellular environments in which they reside. In the present study, we isolated ganglionic eminence neural progenitor cells (geNPC), precursors of the adult striatum, from the ventral forebrain germinal zone and analyzed adhesion, migration, and differentiation of geNPC on various extracellular matrix (ECM) substrates in vitro. Specifically, we evaluated the role of beta1 integrins, a family of cell surface receptors important in neural development, in mediating geNPC behavior on ECM molecules expressed in embryonic brain tissue. Adhesion and migration of geNPC were significantly enhanced on laminin (LN) and fibronectin (FN) relative to other ECM substrates. Antibody perturbation experiments revealed that although geNPC express several beta1 integrins (alpha1beta1, alpha2beta1, alpha3beta1, alpha5beta1, alpha6beta1, alphavbeta1), adhesion and migration on LN and FN were primarily mediated by alpha6beta1 and alpha5beta1, respectively, and these interactions were confirmed by biochemical cross-link/extraction procedures. Finally, neuronal differentiation of geNPC was enhanced on LN, indicating a role for LN in geNPC differentiation. beta1 integrin-ECM interactions may contribute to basic mechanisms of striatal development and may explain the potent migratory capacity of geNPC transplanted into the adult brain. PMID- 15345241 TI - Involvement of Ath3 in CNTF-mediated differentiation of the late retinal progenitors. AB - The cellular diversity of the mammalian retina is underpinned by multipotential neural progenitors that generate retinal neurons and glia with temporal and spatial specificity. It is thought, based on studies using a variety of approaches, that the fate of retinal progenitors is determined through interactions between temporally and spatially arrayed epigenetic cues with intrinsic factors that regulate the competence of cells to respond to such cues. Here, we demonstrate interactions between an intrinsic factor Ath3, a neural bHLH protein, and an extrinsic factor CNTF during the differentiation of the late retinal progenitors along the bipolar cell lineage. Expression of Ath3 is predominantly associated with the late stage of retinal histogenesis when bipolar cells are specified, and in adult it is detected in cells expressing bipolar cell specific markers. We demonstrate that CNTF-induced bipolar cell differentiation is accompanied by an increase in levels of Ath3 transcripts and compromised when Ath3 expression is attenuated. Our study suggests that the influence of CNTF on the differentiation of late retinal progenitors is mediated through Ath3. PMID- 15345242 TI - The transcriptional repressor Mecp2 regulates terminal neuronal differentiation. AB - Rett syndrome (RTT) is a severe neurodevelopmental disorder with features of autism that results from mutation of the gene encoding the transcriptional repressor methyl-CpG binding protein (MECP2). The consequences of loss of a transcription factor may be complex, affecting the expression of many proteins, thus limiting understanding of this class of diseases and impeding therapeutic strategies. This is true for RTT. Neither the cell biological mechanism(s) nor the developmental stage affected by MECP2 deficiency is known. In vivo analysis of the olfactory system demonstrates that Mecp2 deficiency leads to a transient delay in the terminal differentiation of olfactory neurons. This delay in maturation disrupts axonal targeting in the olfactory bulb, resulting in abnormal axonal projections, subglomerular disorganization, and a persistent reduction in glomerular size. These results indicate a critical cell biological function for Mecp2 in mediating the final stages of neuronal development. PMID- 15345243 TI - Axon fasciculation defects and retinal dysplasias in mice lacking the immunoglobulin superfamily adhesion molecule BEN/ALCAM/SC1. AB - The immunoglobulin superfamily adhesion molecule BEN (other names include ALCAM, SC1, DM-GRASP, neurolin, and CD166) has been implicated in the control of numerous developmental and pathological processes, including the guidance of retinal and motor axons to their targets. To test hypotheses about BEN function, we disrupted its gene via homologous recombination and analyzed the resulting mutant mice. Mice lacking BEN are viable and fertile, and display no external morphological defects. Despite grossly normal trajectories, both motor and retinal ganglion cell axons fasciculated poorly and were occasionally misdirected. In addition, BEN mutant retinae exhibited evaginated or invaginated regions with photoreceptor ectopias that resembled the "retinal folds" observed in some human retinopathies. Together, these results demonstrate that BEN promotes fasciculation of multiple axonal populations and uncover an unexpected function for BEN in retinal histogenesis. PMID- 15345244 TI - Gene expression analysis of the critical period in the visual cortex. AB - The development of the primary visual cortex in animals possessing binocular vision is a classical paradigm for the study of activity-dependent neuronal plasticity. To elucidate the genetic determinants of this period of substantial plasticity, we conducted an unbiased and comprehensive transcript profiling analysis with differential display and DNA array techniques. We characterized the transcripts that change significantly between the critical and postcritical periods in the rat binocular visual cortex. We determined if these changes are specific for the visual cortex by simultaneously profiling the hippocampus and examined the impact of sensory experience on the accumulation of the identified transcripts. Our results uncover visual cortex-specific and unspecific transcription programs. Transcripts for protein kinases and phosphatases are particularly regulated. The identified transcripts support the notion that the critical period provides a permissive state for plasticity. PMID- 15345245 TI - [Thrombocytopenia and intensive care unit mortality: a simple marker not to be neglected!]. PMID- 15345246 TI - [No crisis in anaesthesia in France: no shortage of trainees in view]. PMID- 15345247 TI - [Consensus of Societies concerned by emergency medicine]. PMID- 15345248 TI - [Risk factors of mortality in critically ill patients with thrombocytopenia]. AB - OBJECTIVE: Determine predictive factors of intensive care mortality in thrombocytopenic patients. STUDY DESIGN: Retrospective study. PATIENTS AND METHODS: Data including the last 100 patients hospitalised in intensive care unit during period from March 2002 to January 2003 and having presented, at least one time, a platelet count <150 x 10(9)/l. Comparison between patients was realized according to the definitive issue (death or discharge from intensive care). RESULTS: Incidence of thrombocytopenia was 27%. Mortality rate was 53%. Thrombocytopenia in admission was noted in 44% of the patients. IGS II score (OR = 1.05 and p = 0.014), sepsis (OR = 34.2 and p = 0.002) or hepatic dysfunction cases (OR = 42.5 and p = 0.026) were predictive factors of death in intensive care unit when thrombocytopenia occurred. CONCLUSION: Our results concerning prognostic value of thrombocytopenia in intensive care unit are partly similar to those of literature with a surmortality associated to thrombocytopenia related to sepsis and hepatic dysfunction. PMID- 15345249 TI - [Intraoperative arterial hypotension recorded by an anaesthesia information management system]. AB - OBJECTIVES: In order to evaluate the benefits arising from regular recording of intraoperative adverse events, we extracted from our database all episodes of intraoperative hypotension and studied the risk factors of this event. STUDY DESIGN: Retrospective analysis of a large database from two university teaching hospitals evaluating the incidence and the risk factors of intraoperative hypotension by logistic regression. PATIENTS AND METHODS: A data collection chart describing the patient's characteristics, the anaesthetic technique and selected intraoperative incidents was filled for each anaesthetised patient in the operating room and then recorded in the computer database. Data collected in 2001 in patients undergoing general anaesthesia for orthopaedic and general surgery were reviewed and univariate and multivariate analysis were performed using Statview 5.0 and Stata 7.0. RESULTS: Among 11 820 patients who underwent anaesthesia, 2691 were selected. The incidence of intraoperative arterial hypotension was 16.8%. The associated factors were duration of surgery, age and ASA status of the patients. The use of etomidate for induction was not associated with a decreased risk of intraoperative hypotension. CONCLUSION: Systematic recording of intraoperative events in a database has been suggested as useful by many experts for quality-assurance and safety analysis purposes. Analysis of a frequent anaesthesia-related (i.e. hypotension) event did not disclose any relevant factor that might lead to improvement. Running such databases is time consuming and may be expensive. This leads us to question the efficiency of such databases. PMID- 15345250 TI - [French residents in anaesthesia and critical care - a nationwide survey]. AB - OBJECTIVE: We surveyed the residents in their first (R1) and fourth (R4, last) years of residency in anaesthesia and intensive care in France. METHODS: The questionnaires mailed to each resident were designed to obtain personal data, motivation for specializing in anaesthesia and their opinion of their training. RESULTS: The response rates were 48% for the R1 residents and 77% for the R4 residents. There were 40% females in this population and the R1 residents were 25 +/- 1 year old, 29 +/- 2 year-old for the R4 residents. Almost half (46%) of the R1 responders were married as were 74% of the R4 residents. They spent an average of 726 each year on books, computers and other educational items. Most (71%) had opted for anaesthesia after considering other medical specialties, but only 12% had considered surgery. Half (51%) were on a training program that was far away from their home. They thought highly of their training, with the clinical training being rated above the non-clinical component. About half of them had obtained specialized certificates (mainly additional certification in intensive care and antibiotic therapy) during their residency. Two thirds of those in R4 expected to work in a public hospital and about one third expected to work in intensive care. Money was an important factor in their choice of profession. A majority planned to remain in the area where they had graduated in anaesthesia after their residency. Finally, 96% declared that they would choose anaesthesia again if they had to do so. CONCLUSION: French residents in anaesthesia are satisfied of their initial choice for anaesthesia and don't regret it at the end of their residency training. PMID- 15345251 TI - [Disposable material for tracheal intubation: pilot study using Penlon Crystal blades and Penlon Lary Gard sheaths]. AB - OBJECTIVE: The risk of cross infection due to non-conventional infectious agents has led to the development of single-use disposable medical material. For laryngoscopy, two attitudes have been proposed: the use of disposable blades or disposable translucide cover sheaths with steel blades. Practice studies assessing the quality of these devices are scarce, leaving the physician to his own experience. STUDY DESIGN: Open pilot study assessing satisfaction of the users during tracheal intubation. PATIENTS AND METHODS: Hundred patients of general surgery requiring tracheal intubation underwent laryngoscopy in routine condition with disposable blades (Penlon Crystal) or disposable cover sheaths (Penlon Lary Gard) with steel blades. The users gave a general appreciation of the device and specific comments on the easiness of insertion of the blade, intensity of light and its focus, quality of glottic exposure, easiness of tracheal tube insertion, and solidity of the device. RESULTS: With both devices, intubation was impossible in 6% of the patients. The users considered that intubation was performed in comfortable conditions with both devices. Dim light was noticed with Crystal blade. CONCLUSION: In case of use of disposable material, both devices seem to be close in terms of quality of intubation. PMID- 15345252 TI - [Preoperative betablockers use in vascular and thoracic surgery]. AB - OBJECTIVE: The aim of this study was to evaluate betablockers use and effect in the preoperative period in vascular and thoracic surgery. PATIENTS AND METHODS: Retrospective study concerning patients scheduled for high or intermediate surgical risk procedure (n = 321) in a French vascular and thoracic surgery department during 2002. Eligibility for betablocker therapy was defined as previously described for inclusion criteria in Mangano's Study (N Engl J Med 1996;335:1713-20). Therapy effect was evaluated with heart rate in the operating room. RESULTS: Forty-six percent (99/213) of eligible patients for betablockers were really treated in the preoperative period. In patients treated with betablockers, 40% appeared in the operating room with the most favorable heart rate (55-65 b/min). During surgical procedure, heart rate over 80 b/min was observed in more than 30% of patients with betablocker therapy. In patients eligible for betablocker therapy but untreated with these drugs, only 12% have received betablockers in the preoperative period. CONCLUSION: Betablockers underuse is important in the preoperative period, and effect on heart rate is rarely optimal. This fact should encourage physicians to apply protocols for perioperative betablockers administration. PMID- 15345253 TI - [Laboratory animal anaesthesia: influence of anaesthetic protocols on experimental models]. AB - The use of experimental animals requires anaesthesia to provide immobility and analgesia. Animals require anaesthesia not only for ethical reasons but also because pain and stress can alter the quality of research results. Recognition of pain, and its treatment is important throughout the procedure. Before anaesthesia, animals are acclimated and rehydrated. Except in small rodents and in ruminants, in order to avoid vomiting, a fast of 8 to 12 hours before anaesthesia is recommended. In order to protect animals against suffering and distress during transfer, restraint and management, a premedication is administered. Most human anaesthetic products can be used in animals. There are some specific veterinary anaesthetics. Moreover, the anaesthetic effects could be different from specie to an other. In most big animals, induction is realized by intravenous administration. In small rodents, venous puncture and contention could be difficult, and anaesthetic agents may be injected via intraperitoneal or intramuscular way. The principal inconvenient of these administration routes is the impossibility to adjust dose to animal response. In large animals, human anaesthesia material can be used. Some technical adaptations could be necessary in smaller animals. In rodents or in neonatology, specific devices are recommended. ECG, arterial pressure, tidal volume, expired CO(2) and oxygen saturation monitoring assess quality of, and tolerance to anaesthesia. If animals are awaked after anaesthesia, postoperative management is closed to human clinical problems. During animal experimentations, anaesthesia may interact with results. All anaesthetic drugs alter normal physiology in some way and may confound physiologic results. In the literature, most publications do not mention this possible interaction. Investigators need to understand how animals are affected by anaesthetic drugs in order to formulate anaesthetic protocols with minimal effects on data. Extrapolation between different animal species and human and animals about the effects of anaesthetic agents are very hazardous. Great differences exist between the effects observed in vitro and in whole animals. The effects of the anaesthetics could be totally different if they are used alone or in association. The same anaesthetic could have opposite effects from an organ to another. For results validation, the anaesthesia side effects (hypoventilation, hypotension, cooling em leader ) have to be minimized. All new experimental models should require discussing the possible interferences between anaesthesia and results and to compare results obtained with different anaesthetic protocols. PMID- 15345254 TI - [Insertion of catheter for temporary transvenous ventricular pacing at prehospital scene after sudden malfunction of pacemaker]. AB - A 70-year-old patient who had been fitted with a pacemaker for 27 years consulted his cardiologist to check the latest device, which had been changed in 1998. During the examination there was a sudden malfunction of the pacemaker, which became uncontrollable, resulting in an escape rhythm of 15 beats/min and circulatory insufficiency. Attempted treatment by isoprenaline and external transcutaneous systolic stimulation failed, and an intraventricular catheter was therefore positioned successfully as an emergency prehospital manoeuvre. This produced good haemodynamic stability, allowing the transfer of the patient to hospital where his pacemaker was changed. PMID- 15345255 TI - [Prolonged asystole after spinal anesthesia in a patient with Gallavardin's syndrome]. AB - A clinical case of spontaneous ventricular dysrythmia in a 47-year-old patient scheduled for ankle osteosynthesis is reported. During initial peripheral vein canulation, a spontaneous ventricular tachycardia occurred and disappeared spontaneously in about 3 min. It was decided to proceed with surgery. Thirty minutes after spinal anaesthesia, asystole occurred. Normal sinus rhythm was rapidly restored after basic life support. There was no harmful consequence for the patient. He had a history of repetitive monomorphic ventricular tachycardia (Gallavardin type). The aetiologies of asystole after spinal anaesthesia are well known and will be not discussed in the text. Although the origin of the asystole is unclear in this case, the literature on Gallavardin's syndrome is reviewed, showing that a prolonged and complex preoperative assessment is not mandatory in this syndrome. PMID- 15345256 TI - [Pulmonary embolism of cement during vertebroplasty]. AB - A 62-year-old woman with severe osteoporosis experienced pulmonary embolism by polymethylmethacrylate after percutaneous vertebroplasty. The patient immediately developed respiratory and cardiac distress, and a computed tomographic scan revealed the presence of cement in the pulmonary circulation. Proper techniques can minimize the risk of pulmonary embolism during percutaneous vertebroplasty: adequate preparation of cement and fluoroscopy during the procedure are recommended. PMID- 15345257 TI - [Right uncontrollable haemothorax revealing a liver injury with diaphragmatic rupture]. AB - We report a case of a woman with a blunt thoracic trauma and haemorrhagic shock after a road traffic accident. The clinical and complementary examinations revealed an isolated right haemothorax, which was compressive and uncontrollable. The source of bleeding was discovered with delay and during a surgical exploration: it was a liver injury with diaphragmatic rupture but without hepatic herniation and peritoneal effusion. The diagnostic features of blunt diaphragmatic rupture are discussed. PMID- 15345258 TI - [A case of hypertriglycideremia-induced pancreatitis in pregnancy: value of heparin]. AB - We report the case of a 28-year-old secundipari with a history of severe hypertriglycideremia-induced pancreatitis 3 years ago who was admitted in the 37th week of gestation with abdominal pain. A blood sample had a milky aspect and plasma concentrations were as follows: triglycerides 8,5g/l, cholesterol 1000 mg/dl, amylase 574 IU/l, lipase 1310 IU/l. Acute pancreatitis was diagnosed, a caesarean was performed under spinal anaesthesia. The diagnosis was confirmed by CT-scan. Treatment with 15,000 IU heparin per day and intravenous nutrition decreased triglycerides level to less than 1g/l within 48 h. She was discharged 28 days later. Heparin could be a low-cost alternative to plasmapheresis in hypertriglycideremia-induced pancreatitis. PMID- 15345259 TI - "Code blue": monitoring the vital signs of academic anesthesia. PMID- 15345260 TI - The parturient with a scarred uterus, uterine rupture and labor analgesia: facts and fiction. PMID- 15345261 TI - [Assessment of 44 heat and moisture exchange filters. What to choose?]. PMID- 15345262 TI - [Locoregional anesthesia for hand surgery]. PMID- 15345263 TI - [Emergency department policy Implementation and management of the resuscitation room]. PMID- 15345264 TI - Crossing boundaries: implications of advances in basic sciences for the management of addiction. PMID- 15345265 TI - Advances in molecular genetics and the prevention and treatment of substance misuse: Implications of association studies of the A1 allele of the D2 dopamine receptor gene. AB - Substance misuse is influenced by multiple genetic and environmental factors. Recent research has identified a number of potential genetic markers of risk and those associated with drug reward substrates show particular promise. The current study reexamines the extant published data of the association between the D2 dopamine receptor (DRD2) gene minor Taq 1A (A1) allele and substance misuse risk. A series of meta-analyses was performed on 64 studies examining DRD2 A1+ allelic status and substance misuse. In addition, personality was examined as a possible endophenotype. Significant association was found between the A1 allele and severe substance dependence in both Caucasian and non-Caucasian groups. The data did not support a significant association between the A1 allele and personality features. While the specific mechanism underlying these associations requires further elucidation, this genetic marker shows promise as a marker of brain reinforcement processes. Possible ways of utilising the A1 allele to inform prevention and treatment initiatives are discussed. PMID- 15345266 TI - Genes and gene expression in the brain of the alcoholic. AB - Chronic alcoholism leads to localized brain damage, which is prominent in superior frontal cortex but mild in motor cortex. The likelihood of developing alcohol dependence is associated with genetic markers. GABAA receptor expression differs between alcoholics and controls, whereas glutamate receptor differences are muted. We determined whether genotype differentiated the localized expression of glutamate and gamma-aminobutyric acid (GABA) receptors to influence the severity of alcohol-induced brain damage. Cerebrocortical tissue was obtained at autopsy from alcoholics without alcohol-related disease, alcoholics with cirrhosis, and matched controls. DRD2A, DRD2B, GABB2, EAAT2, and 5HTT genotypes did not divide alcoholic cases and controls on N-methyl-d-aspartate (NMDA) receptor parameters. In contrast, alcohol dehydrogenase (ADH)3 genotype interacted significantly with NMDA receptor efficacy and affinity in a region specific manner. EAAT2 genotype interacted significantly with local GABAA receptor beta subunit mRNA expression, and GABB2 and DRD2B genotypes with beta subunit isoform protein expression. Genotype may modulate amino acid transmission locally so as to mediate neuronal vulnerability. This has implications for the effectiveness of pharmacological interventions aimed at ameliorating brain damage and, possibly, dependence. PMID- 15345267 TI - Pleasure into pain: the consequences of long-term opioid use. AB - One consequence of repeated drug administration is the development of adaptations in the nervous system, sometimes termed 'drug-opposite' responses. During administration, the effects of the drug are diminished by these adaptations (tolerance), while cessation of drug use results in the emergence of these drug opposite responses as the withdrawal syndrome. Recent evidence on pain responses challenges this simple notion of withdrawal and suggests that aversive drug opposite states may play a more important role in drug dependence than previously thought. While opioids such as heroin produce analgesia, people with a history of opioid self-administration are hypersensitive to certain kinds of pain during the time they are under the influence of the analgesic drug. This suggests that in pain systems, the drug-opposite response exceeds the pain inhibiting effect of the drug itself. This hyperalgesia is evident in people with a history of heroin use and is not modified by methadone or buprenorphine treatment but is reduced by long-term abstinence from opioids. This same pattern of the drug-opposite response exceeding the drug effect may also occur for mood. While opioids cause elevation of mood, commonly described as euphoria and reduction of emotional distress, methadone maintenance participants show significant negative mood disturbance relative to controls. Thus, for pain and mood, the chronic opioid user under the influence of the drug does not experience an opioid effect diminished by tolerance but a state opposite to the effect of the drug. Increases in drug concentration arising from administration serve only to reduce the degree of pain and mood disturbance. These aversive pain and mood states may contribute to the motivation for continued drug use and the dysfunction associated with drug dependence. PMID- 15345268 TI - Imbalance between neuroexcitatory and neuroinhibitory amino acids causes craving for ethanol. AB - Long-term exposure to ethanol leads to an imbalance in different excitatory and inhibitory amino acids. When ethanol consumption is reduced or completely stopped, these imbalances in different amino acids and neurotransmitters are behaviorally expressed in the form of ethanol withdrawal. Glutamate, a major excitatory amino acid, and GABA, a major inhibitory amino acid, are responsible, at least partly, for ethanol withdrawal symptoms. The hypofunction of GABAA receptors and enhanced function of NMDA receptors are suggested to be responsible for the increase in the behavioral susceptibility during ethanol withdrawal. This imbalance between receptors may be exacerbated by repeated withdrawal. Because multiple and repeated periods of chronic ethanol consumption and withdrawal often occur in alcohol abusers, animal studies on the neurochemical changes in different amino acids following chronic ethanol treatment (CET) that is interrupted by repeated ethanol withdrawal episodes may be of clinical relevance for the development of treatment strategies. Brain glutamate increases during the first cycle of ethanol withdrawal, and this increase is much higher during the third cycle of ethanol withdrawal. The elevated glutamate released in the hippocampus during the first cycle of ethanol withdrawal episode was exacerbated in subsequent withdrawal episodes. Acamprosate, a drug used during human alcohol detoxification, is able to completely block the glutamate increase observed during the first as well as the third withdrawal of ethanol. In ethanol-naive rats, there was no change in the glutamate microdialysate content after an acute ethanol injection. However, when repeated ethanol injections were cued with a vinegar stimulus that had previously been associated with the same ethanol injection, a significant increase in glutamate microdialysate content was assayed. Furthermore, when the cue was omitted, the ethanol injection induced no changes in glutamate microdialysate content in rats that had been previously ethanol conditioned. By comparison, a saline injection had no effect on extracellular glutamate concentration in rats naive for ethanol as well as in rats daily administered with repeated ethanol injections that were not paired with the cue. It appears probable that these conditioned responses by extracellular glutamate concentrations may participate in the environmental cue induced conditioned cravings for ethanol that are thought to be related to the high frequency of relapse in detoxified alcoholics. PMID- 15345269 TI - Rats on the grog: novel pharmacotherapies for alcohol craving. AB - Current pharmacotherapies for alcohol dependence in humans (e.g., naltrexone, acamprosate) are meeting with only limited therapeutic success. The development of novel pharmacotherapies is urgently needed but is reliant upon the screening of large numbers of candidate "anticraving" drugs using appropriate animal models. The development of animal models is complex because (1) laboratory animals are often reluctant to consume large quantities of alcohol, (2) inducing a state of alcohol dependence, analogous to the human condition, may require many months of alcohol exposure, (3) concluding that a given drug selectively reduces alcohol craving requires very carefully controlled experiments, and (4) false positives and false negatives may result from the sometimes distinct physiology and psychology of the alcohol-addicted human and rat. To address some of these problems, our laboratory has recently developed the "beer model" of alcohol dependence and craving. Rats, like humans, have a prodigious appetite for beer and will drink much more beer than equivalent ethanol solutions in water. Beer consumption in rats leads to clear signs of intoxication, anxiety reduction, and signs of withdrawal when beer access is suddenly denied. We have found that beer craving in rats is selectively reduced by the cannabinoid receptor antagonist SR 141716 and the opioid receptor antagonist naltrexone. Combining these two drugs appears to have a synergistic anticraving effect. Other promising pharmacotherapies for the future are discussed. PMID- 15345270 TI - Beating the urge: implications of research into substance-related desires. AB - Despite the advent of improved pharmacological treatments to alleviate substance related desires, psychological approaches will continue to be required. However, the current psychological treatment that most specifically focuses on desires and their management--cue exposure (CE)--has not lived up to its original promise. This paper argues that current psychological approaches to desire do not adequately incorporate our knowledge about the factors that trigger, maintain, and terminate episodes of desire. It asserts that the instigation and maintenance of desires involve both associative and elaborative processes. Understanding the processes triggering the initiation of intrusive thoughts may assist in preventing some episodes, but occasional intrusions will be inevitable. A demonstration of the ineffectiveness of thought suppression may discourage its use as a coping strategy for desire-related intrusions, and mindfulness meditation plus cognitive therapy may help in accepting their occurrence and letting them go. Competing tasks may be used to reduce elaboration of desires, and competing sensory images may have particular utility. The application of these procedures during episodes that are elicited in the clinic may allow the acquisition of more effective strategies to address desires in the natural environment. PMID- 15345271 TI - Negative alcohol consumption outcome associations in young and mature adult social drinkers: a route to drinking restraint? AB - Alcohol 'cognitions' were explored using an implicit methodology [Stacy, Leigh and Weingardt, 1994]. In Study 1, an Associations Questionnaire was developed with young adult undergraduates (median=20 years) comprising culturally available (i.e., high-frequency occurrence) and idiosyncratic (i.e., low-frequency occurrence) positive and negative alcohol consumption outcomes and positive and negative outcomes of behaviors-other-than-alcohol consumption. In Study 2, the relationship was explored between the alcohol consumption of young adult undergraduates (median age = 19 years) and implicit alcohol-related associations made through the Associations Questionnaire. A significant positive relationship was found between consumption and positive and negative culturally available (experimental) outcomes but not for the other two types of (control) outcome. In Study 3, the relationship between alcohol consumption and alcohol-related associations was explored in mature adults (median = 45 years) with many more years' drinking experience. The results of Study 1 were replicated except that a significant positive relationship between consumption and some control outcomes was now found--e.g., negative outcomes of what where hitherto behaviors-other than-alcohol consumption. In Study 4, however, using the same techniques that were used to develop the Associations Questionnaire in Study 1, an extended set of negative alcohol consumption outcomes was found in mature adults (median = 44 years) that included a proportion of the negative outcomes of behaviors-other than-alcohol consumption that had served as controls in Studies 2 and 3. A reanalysis of the data from Study 3, with such items removed from the controls and designated 'new' negative alcohol consumption outcomes, showed a positive relationship between consumption and implicit alcohol-related associations made through these 'new' items of the Associations Questionnaire. The changing profile of associations with negative outcomes of consumption is discussed and related to negative expectancy research and drinking restraint. PMID- 15345272 TI - Reward drive and rash impulsiveness as dimensions of impulsivity: implications for substance misuse. AB - One of the primary personality dimensions or traits that has consistently been linked to substance abuse is impulsivity. However, impulsivity is not a homogenous construct and although many of the measures of impulsivity are correlated, the most recent review of published factor analytic studies has proposed two independent dimensions of impulsivity: reward sensitivity, reflecting one of the primary dimension of J. A. Gray's personality theory, and rash impulsiveness. These two facets of impulsivity derived from the field of personality research parallel recent developments in the neurosciences where changes in the incentive value of rewarding substances has been linked to alterations in neural substrates involved in reward seeking and with a diminished capacity to inhibit behavior due to chronic drug exposure. In this paper, we propose a model that integrates the findings from research into individual differences with recent models of neural substrates implicated in the development of substance misuse. PMID- 15345273 TI - Cognitive and psychological correlates of smoking abstinence, and predictors of successful cessation. AB - The neural circuitry implicated in addictive drug use, which appears to be down regulated in early abstinence, corresponds closely with brain reward pathways. A literature review suggests that responses to incentive stimuli and the ability to inhibit reflexive responses, both of which have been associated with normal functioning in these pathways, might be weakened during acute abstinence from chronic drug use. In an ongoing study, 82 smokers, abstinent overnight before two separate testing occasions, have been assessed after administration of nicotine and placebo lozenges (order of sessions counterbalanced). Nicotine administration is associated with a significant reduction in anhedonia, a near-significant increase in response to financial incentive, enhanced ability to inhibit reflexive eye movements, and increased attentional bias to words with appetitive significance. Fifty-nine participants then initiated a quit attempt and 19 reported relapsing within 7 days. Comparing their performance in the two prequit lozenge assessment sessions, relapsers showed a stronger effect of nicotine on enhancing their ability to inhibit reflexive eye movements and a near-significant trend towards greater nicotine-induced increases in attentional bias toward appetitive words. PMID- 15345274 TI - Biochemical alcohol screening in primary health care. AB - Alcohol biomarkers such as carbohydrate-deficient transferrin (CDT) and gamma glutamyltransferase (GGT) have significant potential for enhancing the quality of medical treatment in primary health care settings. Recent studies demonstrate that these laboratory tests can help the general practitioner in several ways. First, CDT and GGT can detect current heavy drinking in primary care patients with a fair degree of sensitivity (approximately 60% to 70%), with CDT being more specific (approximately 90%). When combined with self-report tests, they can provide a clinically useful alcohol screening battery. Second, elevated CDT and GGT levels have been correlated with specific alcohol-sensitive diseases (e.g., hypertension) and, as such, can serve as risk indicators for those diseases. Third, alcohol biomarkers have proven to be useful in monitoring the effectiveness of brief alcohol interventions with medical patients. Unfortunately, preliminary findings indicate that physicians have little knowledge of current biomarker research as applied to primary health care. Translational studies are needed on methods to facilitate knowledge and use of alcohol biomarkers by general practitioners. PMID- 15345275 TI - Agonist-like, replacement pharmacotherapy for stimulant abuse and dependence. AB - Stimulant abuse and dependence are disproportionately problematic due to the combination of legal and social issues added to the serious behavioural and biological features of the disorders. These problems are compounded by adverse consequences for families and society. Illegality and stigma multiply the consequences of use and difficulties in providing treatment. Specific behavioural interventions have been demonstrated as useful in treatment of substance use disorders (SUDs). Medications also have an important role in treatment. Effective agonist and antagonist pharmacotherapies as well as symptomatic treatments exist for opioid and nicotine dependence. Neither agonists nor antagonists have been approved as uniquely effective for treatment of stimulant abuse or dependence. Still, promising results are emerging for an agonist-like or 'replacement' strategy paralleling that for nicotine and opioid dependence. Supporting data have emerged from both preclinical and clinical research environments. There are scientific, clinical, social, and legal impediments to application of an agonist like approach to stimulant abuse and dependence. Some resemble past and current concerns about opioid replacement. Others are unique to the stimulant agents, effects, and clinical features. Here, the authors consider (1) agonist and antagonist pharmacotherapy strategies; (2) preclinical research, including methodological approaches, opioid and nicotine replacement, and agonists for stimulant dependence; (3) clinical reports with stimulant medications in cocaine dependence, and the amphetamine replacement strategy for amphetamine dependence; (4) application of agonist-like/replacement strategies, including clinical requirements and risks; and (5) directions for research. PMID- 15345276 TI - Topiramate-induced neuromodulation of cortico-mesolimbic dopamine function: a new vista for the treatment of comorbid alcohol and nicotine dependence? AB - Alcohol and nicotine dependence are commonly occurring disorders that together represent the most important preventable causes of morbidity and mortality in the United States. While there have been differences of opinion as to which disorder to treat first when they occur, there is growing evidence that a management strategy addressing both conditions contemporaneously would be optimal. Advances in the neurosciences have demonstrated not only that the reinforcing effects of both alcohol and nicotine are mediated by similar mechanisms resulting in enhanced activity of the cortico-mesolimbic dopamine system, but that their neurochemical interactions can lead to an aggregation of these effects. Despite this striking neurobiological overlap between alcohol and nicotine consumption, few studies have sought to take advantage of this commonality by devising a pharmacological approach that serves to treat both disorders. The results of our proof-of-concept study showed that topiramate is a promising medication for the treatment of both alcohol and nicotine dependence, presumably by its ability to modulate cortico-mesolimbic dopamine function profoundly; however, other mechanisms might also contribute to this effect. Further studies are ongoing to establish and extend topiramate's efficacy in the treatment of each and both disorders. PMID- 15345277 TI - Neuroscience research on the addictions: a prospectus for future ethical and policy analysis. AB - The increasing evidence that many addictive phenomena have a genetic and neurobiological basis promises improvements in societal responses to addiction that raise important ethical and social policy issues. One of the major potential benefits of such research is improved treatment of drug addiction, but in order to do the research required to realize this promise, it will be necessary to address ethical doubts raised about the capacity of addicted persons to give free and informed consent to participate in studies that involve the administration of drugs of dependence. Neuroscience research on addiction promises to transform the long running debate between moral and medical models of addiction by providing a detailed causal explanation of addiction in terms of brain processes. We must avoid causal models of addiction being misinterpreted as supporting simple-minded social policies, e.g., that we identify the minority of the community that is genetically and biologically vulnerable to addiction and hence can neglect social policy options for reducing addiction, including drug control policies. Causal accounts of addiction supplied by neuroscience and genetic research may also be seen to warrant the use of pharmacotherapies and drug vaccines under legal coercion. Neuroscientists also need to anticipate the ethical issues that may arise if the knowledge that they produce delivers interventions that enhance human cognitive and other capacities. Advances in neuroimaging that enable us to identify "addicts" or predict future risk of addiction will raise concerns about invasion of privacy, third-party use of neuroimaging data, the powers of courts to coerce defendants to undergo such tests, and consumer protection against the overinterpretation of test results. Given the strong public and media interest in the results of their research, neuroscientists and geneticists have a moral obligation, and a professional interest, to minimize popular misunderstandings of their work in the media that may rebound to its detriment. PMID- 15345278 TI - From basic research to public health policy: WHO report on the neuroscience of substance dependence. AB - The World Health Organization (WHO) report on the Neuroscience of Psychoactive Substance Use and Dependence represents a summary of the latest scientific knowledge on the role of the brain in substance dependence. The findings of the report have important applications to both interventions and public health policy. Specifically, the report notes that psychoactive substances have physical and symbolic effects that are rewarding to the user. It notes that adverse consequences of psychoactive substance use are related to the level, pattern, mode, and context of use. Dependence is seen as a complex disorder involving brain mechanisms rather than a failure of will. The report notes that psychoactive substance use often coexists with other mental health problems, and that stigma associated with substance misuse inhibits treatment seeking. There is growing evidence for the efficacy and effectiveness of interventions, including advances in pharmacotherapy. However, these advances raise ethical issues that need to be addressed. PMID- 15345279 TI - Harnessing zebrafish for the study of white blood cell development and its perturbation. AB - Considerable progress has been made in understanding the molecular basis of normal white blood cell development and its perturbation in disease through the use of clinical studies and traditional animal and cell line models. Despite this, however, many questions are still being answered and white blood cell disorders, including leukemia and lymphoma, remain a significant health problem. The zebrafish (Danio rerio) has emerged as a powerful alternative vertebrate model for the study of development and disease. We review the recent application of zebrafish to the study of white blood cell development and its disruption, particularly leukemogenesis. Such studies have highlighted the overall conservation of these processes throughout vertebrates, and establish zebrafish as a useful experimental model. This organism is now poised to make an important contribution to our understanding of the underlying genetic control of white blood cell development and its disruption, as well as the identification of new therapeutic agents. PMID- 15345281 TI - Transcript profile of CD4+ and CD8+ T cells from the bone marrow of acquired aplastic anemia patients. AB - OBJECTIVE: Immune-mediated destruction of hematopoietic stem and progenitor cells is pathophysiologic in most cases of aplastic anemia (AA). We have successfully determined the gene expression profile of the marrow CD34+ target cells in AA. T cells producing IFN-gamma and TNF-alpha have been implicated in hematopoietic destruction in AA. We sought to characterize T cells as immune mediators using the microarray approach. MATERIALS AND METHODS: We applied Affymetrix GeneChip techniques to determine the detailed profile of mRNA expression of CD4+ and CD8+ cells from the BM of newly diagnosed AA patients and healthy volunteers. For validation, we confirmed our microarray results using quantitative real-time PCR. RESULTS: Compared to healthy controls, there were 178 and 183 differentially expressed genes in patients' CD4+ cells and CD8+ T cells, respectively; activities of 22 selected genes were confirmed using real-time PCR. Dysregulated genes included those encoding cytokines/chemokines, and involved in transcription regulation, calcium and ion channel formation, and cell adhesion. Unexpected findings were overexpression of toll-like receptor genes in marrow CD4+ cells of patients and of genes for killer-cell immunoglobulin-like receptors (KIR) in AA marrow CD8+ cells. CONCLUSIONS: Our detailed results at the mRNA level provide insights into the mechanism of AA. Both innate and adaptive immune responses of CD4+ and CD8+ T cells appear to be active in immune-mediated marrow destruction. A variety of cytokines and chemokines active in pathophysiologic cells likely play important roles in the recruitment and activation of lymphocytes to cytotoxic effectors for marrow hematopoietic target cells in AA. PMID- 15345280 TI - A new interferon, limitin, displays equivalent immunomodulatory and antitumor activities without myelosuppressive properties as compared with interferon-alpha. AB - OBJECTIVE: Limitin is a new member of type I interferon (IFN) identified with an expression cloning based on the growth suppression of a myelomonocytic leukemia cell line WEHI3. Although limitin uses the IFN-alpha/beta receptor, its signal transduction pathways to express the antiviral effects are different from those of IFN-alpha. To clarify the characteristics of limitin, we compared the biological activities of limitin, such as the antiviral, immunomodulatory, antitumor, and myelosuppressive effects, with IFN-alpha. MATERIALS AND METHODS: Limitin and IFN-alpha were titered with a cytopathic effect dye binding assay. Induction of MHC class I on a keratinocyte cell line PAM212 was estimated with flow cytometry. Induction of OVA-restricted cytotoxic T lymphocyte (CTL) activity was analyzed with 51Cr release assay. Antiproliferative effects were evaluated with 3H-thymidine incorporation assay using WEHI3 and a lymphoblast cell line L1210. Myelosuppresive effects were evaluated with colony assay. In vivo side effects were estimated after the injection of limitin or IFN-alpha. RESULTS: Limitin had relatively higher antiviral activity than IFN-alpha. Limitin induced the surface expression of MHC class I, the enhancement of CTL activity, and the growth inhibition of lymphohematopoietic cell lines as strong as IFN-alpha. Nevertheless, the treatment of mice with limitin showed neither myelosuppression nor fever that are common adverse effects of IFN-alpha. CONCLUSIONS: Strong immunomodulatory, antitumor, and antiviral effects with weak myelosuppressive and weak acute toxic effects of limitin indicate that it may be useful as a new therapeutic drug for virus-hepatitis and cancers. PMID- 15345282 TI - Fanconi anemia type C-deficient hematopoietic cells are resistant to TRAIL (TNF related apoptosis-inducing ligand)-induced cleavage of pro-caspase-8. AB - OBJECTIVE: The pathophysiology of bone marrow failure in Fanconi anemia (FA) patients is thought to involve excessive apoptosis involving signaling triggered by fas ligation and tumor necrosis factor (TNF)-alpha, or interferon (IFN)-gamma exposure. We investigated whether a new member of the TNF family, TRAIL (TNF related apoptosis-inducing ligand), would similarly trigger preferential apoptotic cell death in FA phenotype cells. MATERIAL AND METHODS: Hematopoietic cells from FANCC(-/-) transgenic mice and human FA-C lymphoblasts (HSC536N) as well as their phenotypically corrected counterparts (FANCC(+/+), HSC536/FA-Cneo) were compared for their response to apoptosis induction by TRAIL and fas ligation in the presence or absence of IFN-gamma. Cells were also studied for the protein and gene expression of TRAIL-receptors, caspase-8 and its inhibitory protein, FLIP. RESULTS: TRAIL exposure by itself or in combination with IFN-gamma did not lead to preferential apoptosis induction in human and murine FA-C phenotype hematopoietic cells. This resistance was unrelated to the expression of TRAIL receptors or FLIP isoforms, but correlated with absent cleavage of pro-caspase-8. Results were validated by those from gene expression profiling of relevant genes in the two lymphoblast cell lines. CONCLUSION: TRAIL, in contrast to fas ligation, does not induce preferential apoptosis in FA-C phenotype cells despite shared downstream signaling described in non-FA models. These data provide further insight into the complexity of FA-C-regulated apoptotic signaling. PMID- 15345283 TI - Deterministic dynamics control oscillations of bone marrow cell proliferation. AB - OBJECTIVE: The production of blood cells in vivo, both normal and tumoral, displays oscillatory dynamics. Many cells in long-term cultures also show large amplitude oscillations of proliferative rate. Therefore we examined the proliferation dynamics of mouse bone marrow cells (MBM) and their clonogenic progenitor production (BMP), in order to characterize these dynamics. METHODS: Five Dexter-type cultures of MBM cells and their clonogenic BMP production were examined for up to seven-months periods of time. The recorded time series exhibited a complex pattern of oscillations with variable amplitudes. We previously reported a method that allowed analysis of such nonlinear dynamics of hepatoma cell proliferation. We applied this method, based on the two-dimensional recurrent representation of data, to analyze the fluctuations of bone marrow cells proliferation. RESULTS: The proliferation rate of mouse bone marrow cells shows large amplitude oscillations every 2 to 3 weeks. Mathematical analysis revealed a deterministic mechanism that controls all proliferation local maxima of MBM cells. Dynamics for progenitor production resembled that of parental cells. This reflects a predominant negative feedback on bone marrow cell proliferation. CONCLUSION: These dynamics were opposite of that previously described for hepatoma cells where the dominant control is applied to the local minima (troughs of proliferation). Therefore, the complex system of cell proliferation is controlled by a bipolar mechanism, with a predominant dampening command depending on the cell type. We propose that the dominant dampening control of local maxima in bone marrow cells protects the stock of stem cells. PMID- 15345284 TI - DNA microarray analysis of dysplastic morphology associated with acute myeloid leukemia. AB - OBJECTIVE: Acute myeloid leukemia (AML) develops de novo or secondarily to either myelodysplastic syndrome (MDS) or anticancer treatment (therapy-related leukemia, TRL). Prominent dysplasia of blood cells is apparent in individuals with MDS related AML as well as in some patients with TRL or even with de novo AML. The clinical entity of AML with multilineage dysplasia (AML-MLD) is likely to be an amalgamation of MDS-related AML and de novo AML-MLD. The aim of this study was to clarify, by the use of high-density oligonucleotide microarrays, whether these subcategories of AML are intrinsically distinct from each other. MATERIALS AND METHODS: The AC133+ hematopoietic stem cell-like fractions were purified from the bone marrow of individuals with de novo AML without dysplasia (n = 15), AML-MLD (n = 11), MDS-related AML (n = 11), or TRL (n = 2), and were subjected to the synthesis of cRNA which was subsequently hybridized to microarray harboring oligonucleotide corresponding to more than 12,000 probe sets. RESULTS: We could identify many genes whose expression was specific to these various subcategories of AML. Furthermore, with the correspondence analysis/three-dimensional projection strategy, we were able to visualize the independent, yet partially overlapping, nature of current AML subcategories on the basis of their transcriptomes. CONCLUSION: Our data indicate the possibility of subclassification of AML based on gene expression profiles of leukemic blasts. PMID- 15345285 TI - The comparative analysis of serum proteomes for the discovery of biomarkers for acute myeloid leukemia. AB - OBJECTIVE: Acute myeloid leukemia (AML) develops as the consequence of a series of genetic changes in a hematopoietic precursor cell. However, the definitive diagnostic protein biomarkers for AML are still unclear. In our study to identify the biomarkers for an initial diagnosis, detection of relapse, and monitoring the minimal residual disease in AML by a less invasive method, serum proteins reflecting alterations in their proteomes were analyzed. MATERIALS AND METHODS: We compared the two-dimensional electrophoresis patterns of human sera of 12 patients with AML with those of 12 normal subjects. The differentially expressed spots were identified by matrix-assisted laser desorption/ionization time-of flight and electrospray ionization quadupole time-of-flight mass spectrometries. RESULTS: Eight proteins that expressed differentially in the AML group were found. The expression levels of alpha-2-HS-glycoprotein, complement-associated protein SP-40, 40, RBP4 gene product, lipoprotein C-III, and an unknown protein were downregulated in serum of AML patients, whereas the other three proteins, including immunoglobulin heavy-chain variant, proteosome 26S ATPase subunit 1, and haptoglobin-1 were upregulated. CONCLUSION: These results suggest that these proteins can be used as less invasive diagnostic and monitoring biomarkers of AML if further studies are done. PMID- 15345286 TI - Hematopoietic activity of common marmoset CD34 cells isolated by a novel monoclonal antibody MA24. AB - OBJECTIVE: We focused on a small New World monkey, the common marmoset (Callithrix jacchus), to establish a nonhuman primate model of the treatment of hematological disorders. In this study, we developed the first monoclonal antibodies (MAbs) against marmoset CD34 and tested the in vitro and in vivo hemopoietic activity of cell populations isolated using one of these MAbs. METHODS AND RESULTS: Marmoset cDNA encoding a human CD34 homologue was cloned from bone marrow (BM)-derived RNA using reverse transcription polymerase chain reaction and rapid amplification of cDNA ends. The amino acid sequence of the marmoset CD34 had 81% homology with the human sequence. Five mouse MAbs were raised against marmoset CD34 transfectant. One representative MAb, MA24 (IgM), reacted with approximately 0.5 to 1% of BM mononuclear cells (MNCs), where the colony-forming unit granulocyte/macrophage (CFU-GM) was enriched approximately 11 to 75-fold as compared with the whole BM MNCs. Multilineage differentiation of marmoset CD34+ cells in NOD/SCID mice was confirmed by flow cytometry 1 month after xenotransplantation. CONCLUSION: These results demonstrated that MA24 is useful for the analysis and enrichment of hematopoietic progenitor cells in the marmoset model for preclinical experiments. PMID- 15345287 TI - Mutant N-ras preferentially drives human CD34+ hematopoietic progenitor cells into myeloid differentiation and proliferation both in vitro and in the NOD/SCID mouse. AB - OBJECTIVES: Ras oncogene mutations are the most frequently observed genetic abnormality (20-40% of patients) in acute myeloid leukemia (AML), and in the preleukemic conditions myelodysplastic syndrome (MDS) and myeloproliferative disorder (MPD). We have previously shown that mutant N-ras (N-rasm) can induce myeloproliferative disorders and apoptosis in a murine reconstitution system. In the present study we investigated the effect of N-rasm in human primary hematopoietic progenitor cells (HPC). METHODS: Cord blood CD34+ hematopoietic progenitor cells (HPC) were transduced with retroviral vectors containing green fluorescence protein (GFP) alone, or in combination with N-rasm. Cells were then cultured in vitro with a cytokine supplement or cocultured with murine stroma MS 5 cells. The in vivo behavior of transduced cells was examined in the NOD/SCID mouse model. RESULTS: N-rasm-transduced cells exhibited greater proliferative capacity; a higher frequency of granulocyte-macrophage colony-forming unit (CFU GM); and an increase in myelomonocytic lineage cells with a concomitant decrease in lymphoid and erythroid cells. Analysis of transduced HPC in NOD/SCID mice revealed higher bone marrow engraftment by N-rasm HPC and increased numbers of myeloid lineage cells. CONCLUSIONS: The results demonstrate that N-rasm in HPC induces myeloproliferation both in vitro and in the NOD/SCID mouse model as a primary event that does not appear to be dependent on cooperating transforming events. PMID- 15345288 TI - Allogeneic bone marrow-derived flk-1+Sca-1- mesenchymal stem cells leads to stable mixed chimerism and donor-specific tolerance. AB - OBJECTIVE: To investigate the possibility of flk-1+Sca-1- bone marrow-derived mesenchymal stem cells (bMSCs) to induce stable mixed chimerism and donor specific graft tolerance. METHODS: Allogeneic flk-1+Sca-1- bMSCs and syngeneic bone marrow (BM) cells were cotransplanted into lethally irradiated (8.5 Gy) recipient mice. FACS was used to analyze the chimerism 150 days later. Donor-type skin transplantation was performed to observe donor-specific immunotolerance in recipient mice. Mixed lymphocyte reaction (MLR) and mitogen proliferative assays were performed to evaluate proliferative response of splenocytes from recipient mice. RESULTS: More than 5% donor-derived CD3+ cells were detected in splenocytes of recipient mice. Long-term survival of donor-type skin grafts was observed. MLR and mitogen proliferative assays showed that recipient mice had low immunoresponse to donor cells but retained normal ConA-induced proliferative response compared with normal mice. CONCLUSION: Our results show for the first time that induction of stable mixed hematopoietic chimerism can be achieved with allogeneic flk-1+Sca-1- bMSC transplantation, which leads to permanent donor specific immunotolerance in allogeneic host and results in long-term allogeneic skin graft acceptance. PMID- 15345289 TI - Circulating hematopoietic stem cells do not efficiently home to bone marrow during homeostasis. AB - OBJECTIVE: Hematopoietic stem cells (HSC), normally resident in bone marrow, can be detected in the murine and human circulation. It is thought that HSC move in and out of bone marrow daily and that returning HSC are generally equivalent to their bone marrow counterparts in phenotype and function. However, large numbers of mononuclear blood cells are required to rescue animals from lethal irradiation, indicating either that the prevalence of circulating HSC is low, or they are inherently deficient in their repopulating ability. Accordingly, recent data suggest that circulating HSC may be unable to stably engraft WBM under homeostatic conditions. The purpose of this study was to explore these dynamics in detail using parabiosis and bone marrow transplantation. MATERIALS AND METHODS: The WBM and skeletal muscle HSC stem cell compartments of parabiosed CD45 congenic mice were analyzed functionally (via bone marrow transplantation) and phenotypically (via flow cytometry) for circulating stem cells at specific time points postparabiosis and after surgical separation. RESULTS: Surprisingly, we find that stem cells trafficking out of bone marrow and into the circulation do not stably return to bone marrow, although long-lived lymphoid precursors do stably re-engraft. Circulating HSC do, however, take up residence in skeletal muscle, wherein they account for HSC activity. CONCLUSION: Circulating HSC are not in flux with the bone marrow HSC and can persist in peripheral tissues. PMID- 15345290 TI - Evidence for megakaryocyte engraftment following reduced-intensity conditioning. AB - Assessment of donor chimerism is becoming increasingly important in patients undergoing reduced-intensity conditioning (RIC) allogeneic bone marrow transplants, due to the possibility of mixed chimeras. This regimen has been used successfully for patients with leukemia and genetic disorders with donor chimerism occurring in the myeloid, lymphoid, and/or erythroid lineages. Less toxic RIC expands the potential application of stem cell transplants to patients with nonmalignant disorders of hematopoiesis, such as the severe form of Glanzmann thrombasthenia, who previously were not considered suitable candidates based on risk-benefit analysis. To assess megakaryocyte/platelet chimerism after stem cell transplantation conducted with RIC, we used restriction fragment length polymorphism (RFLP) and sequence analyses of the HPA-3 polymorphism in the megakaryocyte/platelet-specific glycoprotein alphaIIb. In this study we show that at 23 weeks post-RIC, a leukemia patient acquired the HPA-3 donor phenotype at the DNA and platelet RNA levels. PMID- 15345291 TI - Large-scale methacrylate monolithic columns: design and properties. AB - Monoliths represent a special class of chromatographic supports. In contrast to other stationary phases, they consist of a single piece of highly porous material through which a sample is mainly transported by convection. As a consequence, monoliths enable fast separations and exhibit flow-unaffected properties, which make them attractive for purification of macromolecules like proteins or DNA. In this work, methacrylate-based monolithic columns with the bed volume up to 8000 ml are characterized. They perform high-resolution separations of several hundreds of grams of proteins per hour by utilizing liter per minute flow rates. They are incompressible under these operating conditions and resistant to strong alkaline conditions. PMID- 15345292 TI - Analysis of amino acid-carbohydrate mixtures by anion exchange chromatography and integrated pulsed amperometric detection. AB - A review is presented of recent developments in the area of analysis of amino acid-carbohydrate mixtures. Based on its broad selectivity, the AminoPac PA10 column exhibits a remarkable capability to perform simultaneous separations of amino acids and carbohydrates. This ability is further enhanced by the equal sensitivity for carbohydrates and amino acids exhibited by the "amino acid" integrated pulsed electrochemical detection (IPAD) waveforms. Equimolar levels of carbohydrates and amino acids are separated either by optimized elution gradients alone or by a combination of modified gradients and a bi-modal IPAD waveform. Samples containing large amounts of carbohydrates may be analyzed after suitable sample preparation. Both a manual off-line method and a fully automatic on-line method are discussed. In addition, we will review the application of these methods to various types of samples, including cell culture media, glycoprotein hydrolysates, beverages, condiments and soil extracts. PMID- 15345293 TI - Recent developments in electrolytic devices for ion chromatography. AB - Recent developments in new electrolytic devices that utilize the electrolysis of water and charge-selective electromigration of ions through ion-exchange media have significantly changed the routine operation of ion chromatographic methods. Examples of these new electrolytic devices include on-line eluent generators that produce high-purity electrolyte eluents using deionized water as the carrier stream, continuously regenerated trap columns that remove ionic contaminants in the eluents, and continuously regenerated suppressors that reduce eluent background conductance prior to conductivity detection. The combined use of these electrolytic devices has made it possible to perform various ion chromatographic separations using only deionized water as the mobile phase. This paper reviews the operation principles of these electrolytic devices and their applications in the ion chromatographic determination of anionic and cationic analytes. PMID- 15345294 TI - Instrumentation for advanced microseparations in pharmaceutical analysis and proteomics. AB - Small-volume chromatographic columns are only able to generate narrow peaks when flow rates, injection volume and instrument components, such as detector, connecting tubing and fittings, are matched to the peak dispersion from the column. Criteria for the proper design of chromatographic instrumentation are therefore derived from a general model on total dispersion. The performance of such a system is then experimentally evaluated from applications run on narrow bore, small-volume columns. In order to achieve flow rates that match the dimensions of such columns, a new concept for electronic flow control (EFC) is introduced. A theoretical optimization of column efficiency and throughput is discussed and the results verified with practical examples on short, narrow-bore columns packed with small, porous and superficially porous particles. For complex sample mixtures, the concept of peak capacity is introduced and applied to orthogonal separation principles in multiple chromatographic dimensions through column switching techniques. PMID- 15345295 TI - Peptide separation by Hydrophilic-Interaction Chromatography: a review. AB - Recent developments in the separation of peptides by high-performance liquid chromatography (HPLC) using polar sorbents with less polar eluents are summarized in this review. This separation mode is now commonly referred to as Hydrophilic Interaction Chromatography (HILIC). The retention mechanism and chromatographic behavior of polar solutes under HILIC conditions are studied on TSKgel Amide-80 columns, which consist of carbamoyl groups bonded to a silica gel matrix, using a mixture of acetonitrile (MeCN)-water containing 0.1% trifluoroacetic acid (TFA). Some applications are given in peptide field using Hydrophilic-Interaction Chromatography. PMID- 15345296 TI - Comparison of selected analytical techniques for protein sizing, quantitation and molecular weight determination. AB - Protein analysis techniques are developing fast due to the growing number of proteins obtained by recombinant DNA techniques. In the present paper we compare selected techniques, which are used for protein sizing, quantitation and molecular weight determination: sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), lab-on-a-chip or microfluidics technology (LoaC), size exclusion chromatography (SEC) and mass spectrometry (MS). We compare advantages and limitations of each technique in respect to different application areas, analysis time, protein sizing and quantitation performance. PMID- 15345297 TI - Process control and drug analysis with an on-line capillary electrophoresis system. AB - Inorganic and organic ions in pulp and paper process waters and drug mixtures were studied. Real-time measurements and on-line simulated analyses were made. A novel on-line capillary electrophoresis instrument equipped with a fixed wavelength UV detector having a 254-nm filter was used for the analyses. Three dynamic sample and electrolyte micromodules provided on-line sampling, sample introduction and solution feedings. The system was used to monitor process samples containing thiosulphate, chloride, sulphate, oxalate, acetate and carbonate. It approved good performance in separation of ethacrynic acid, furosemide, probenecid, bumetanide and hydrochlorothiazide as well as that of normethanephrine, methanephrine and dopamine standard mixtures. The LOD and LOQ values of the ions and drugs ranged from 0.1 to 1 mg/l for LOD and from 1 to 10 mg/l for LOQ. Sensitivity of the drugs was higher due to the single wavelength available in the system. It required that the drugs were to be identified by indirect UV detection. Repeatability of the analyses was good (RSD% below 5). PMID- 15345298 TI - CarboPac PA20: a new monosaccharide separator column with electrochemical detection with disposable gold electrodes. AB - This report documents the development of a new monosaccharide separator column (CarboPac PA20, 3x150 mm) that allows fast, efficient monosaccharide separations with good spacing. It is based on a new chemistry with a reduced resin particle size (from 10 to 6.5 microm). Faster, more efficient separations of glycoprotein monosaccharides with better spacing were achieved across a range of isocratic NaOH concentrations at lower flow rates. Detection sensitivity was improved, enabling routine low to sub pmol monosaccharide determinations. Glycoprotein monosaccharides eluted in less than 10 min at a flow rate of 0.5 ml/min. Furthermore, when used with an AminoTrap guard column, the protein matrix consisting of amino acids and peptides (released by acid hydrolysis of glycoprotein) did not interfere with monosaccharide analysis. Compared to previous CarboPac columns (CarboPac PA1 and CarboPac PA10), the CarboPac PA20 has improved selectivity with respect to glycoprotein monosaccharides. The improved selectivity results in better separation of glucosamine and galactose, enabling the accurate determination of monosaccharide ratios for undergalactosylated glycoproteins. Finally, disposable gold working electrodes that eliminate the possibility of working electrode recession affecting peak area response were used. PMID- 15345299 TI - Applications and performance of a MALDI-ToF mass spectrometer with quadratic field reflectron technology. AB - A new matrix-assisted laser desorption/ionization time of flight mass spectrometer (MALDI-ToF MS), developed specifically for the identification and characterization of proteins and peptides in proteomic investigations, is described. The mass spectrometer which can be integrated with the 2-D gel electrophoresis workflow is a bench-top instrument, enabling rapid, reliable and unattended protein identification in low-, as well as high-throughput proteomics applications. To obtain precise information on peptide sequences, the instrument utilizes a timed ion gate and a unique quadratic field reflectron (Z2 technology), allowing single-run, post-source decay (PSD) of selected peptides. In this study, the performance of the instrument in reflectron, PSD and linear mode, respectively, was investigated. The results showed that the limit of detection for a single peptide in reflectron mode was 125 amol with a signal to noise ratio exceeding 20. Average mass resolution for peptides larger than 2000 u was around 13,000 full width, half maximum (FWHM). The limit for protein identification during peptide mass fingerprinting (PMF) was 500 amol with a sequence coverage of 18%. Mass error during PMF analysis was less than 15 ppm for 17 out of 25 (68%) identified peptides. In PSD mode, a complete series of y-ions of a CAF-derivatized peptide could be obtained from 3.75 fmol of material. The average mass error of PSD-generated fragments was less than 0.14 u. Finally, in linear mode, intact proteins with molecular masses greater than 300,000 u were detected with mass errors below 0.2%. PMID- 15345300 TI - The methodology for determining the efficacy of antibody-mediated immunity. AB - The basic method for evaluating the efficacy of antibody-mediated immunity (AMI) dates from the 1890s and involves the administration of specific Ab to an immunologically naive host followed by microbial or toxin challenge. Other methods used to evaluate AMI involve correlating the presence of specific Ab with resistance to microbial disease and associating susceptibility to certain microbes with host immunoglobulin deficits. Unfortunately, each method has theoretical and practical problems that limit their usefulness when negative results are obtained. The application of hybridoma technology to investigate the efficacy of AMI has shown that it is possible to generate protective monoclonal antibodies even to microbes for which the standard methodologies indicate no role for AMI. Furthermore, studies with monoclonal antibodies suggest various explanations for the inability of standard methods to demonstrate the potential efficacy of AMI for certain pathogens. PMID- 15345301 TI - The development and application of a surface plasmon resonance-based inhibition immunoassay for the determination of warfarin in plasma ultrafiltrate. AB - Warfarin is the most widely prescribed oral anticoagulant for the management of a wide variety of thromboembolic disorders such as atrial fibrillation and deep vein thrombosis. A panel of warfarin-protein conjugates were produced and characterised and subsequently used for the production of monoclonal antibodies to warfarin. Following characterisation, the monoclonal antibodies were used in the development of a surface plasmon resonance-based inhibition immunoassay for the determination of the physiologically active 'nonprotein'-bound fraction of the drug in plasma ultrafiltrate. The inhibition immunoassay was compared with an existing high-performance liquid chromatography (HPLC) chromatographic technique for the determination of warfarin in plasma ultrafiltrate, and an excellent correlation was achieved between the two independent analytical techniques. The ligand-binding capacity and stability of various immobilised ligands were also compared. The BIACore-based inhibition immunoassay demonstrated an assay precision range of approximately 4-250 ng/ml, which is within the clinical range and demonstrated good reproducibility and robustness. PMID- 15345302 TI - Validation and comparative analysis of a multiplexed assay for the simultaneous quantitative measurement of Th1/Th2 cytokines in human serum and human peripheral blood mononuclear cell culture supernatants. AB - There is increasing evidence suggesting a relationship between cytokine levels and disease pathogenesis, which has led to interest in analyzing multiple cytokines in biological fluids and culture supernatants for various research and clinical studies. The introduction of methodologies allowing simultaneous measurement of interrelated biomarkers/cytokines has further revolutionized this process. In contrast to tissue culture supernatant, the measurement of cytokines in serum has proven to be difficult to characterize in multiplexed formats because of the presence of large dynamic concentration ranges of proteins and other interfering factors that are present in this matrix. In the present study, we have used the microsphere-based multiplex method to simultaneously quantitate and compare six analytes, encompassing a representation of the Th1/Th2 cytokine panel (interleukin (IL)-2, IL-4, IL-5, interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and IL-10), in both serum and culture supernatants from peripheral blood mononuclear cells (PBMCs). A detailed validation procedure for these determinations is described along with a comparative analysis of the performance of the multiplexed assay in serum and culture supernatant matrices. Our results indicate that precision of the multiplexed assay is comparable in both culture supernatant and serum. However, the accuracy of quantification of cytokines in the serum matrix but not in culture supernatant may be compromised depending upon the cytokine being analyzed. Therefore, one must use caution when interpreting data from such complex matrices. Nevertheless, this assay format is appropriate to profile cytokines in clinical trial samples. PMID- 15345303 TI - A novel, rapid and sensitive heterotypic cell adhesion assay for CD2-CD58 interaction, and its application for testing inhibitory peptides. AB - The immunoglobulin CD2 is a cell adhesion molecule that mediates T-cell activation by binding to its receptor CD58 on antigen-presenting cells (APCs). Modulation or inhibition of this interaction has been shown to be therapeutically useful. E-rosetting assay is usually applied in the study of the modulation of CD2-CD58 interaction. In this study, we demonstrated a novel, rapid and sensitive heterotypic cell adhesion assay for CD2-CD58 interaction. The CD2 expression on the surface of Jurkat cells and the CD58 expression on the Caco-2 cells were confirmed by flow cytometry and ELISA studies, respectively. Then Jurkat cells were fluorescent-labeled with 2 microM of BCECF-AM for 45 min at 37 degrees C before adding to confluent Caco-2 monolayers cultured in 96-well culture dishes. After 30 min, non-adherent Jurkat cells were removed by washing with PBS, while the monolayer-associated Jurkat cells were lysed with 0.5 ml of 2% Triton X-100 in 0.1 M NaOH. Fluorescence (FL) was quantitated using a microplate fluorescence analyzer with BCECF's excitation maximum of 485 nm and emission maximum of 535 nm. This method was successfully applied for testing inhibitory peptides to CD2 CD58 interaction. PMID- 15345304 TI - A robust human T-cell culture method suitable for monitoring CD8+ and CD4+ T-cell responses from cancer clinical trial samples. AB - Many tumor antigenic determinants have been identified and included in cancer clinical trials. Due to low T-cell frequencies even after vaccination, few T-cell responses can be revealed ex vivo without in vitro stimulation. Various expansion protocols have been employed for this purpose and the outcomes tend to be quite variable, partly due to the high complexity involved in the protocols. Here we systematically studied various common culture conditions including sera, cytokines and feeders and describe a reliable "bulk" culture method that is robust, simpler and more economical. We demonstrated that fetal calf serum (FCS) supported T-cell proliferation better than multiple commercially available pooled human AB sera. IL-2 is critical in our cultures, but IL-7, IL-15 and anti-CTLA-4 in combination with IL-2 did not further enhance T-cell expansion. We typically achieve more than a 40-fold expansion within a 10-day culture period for antigen specific T cells measured by HLA-peptide tetramer before and after culture. This method was not only validated by multiple operators as a standard operating procedure for monitoring T-cell responses but was also successfully used for discovering novel CD8+ and CD4+ T cells specific to previously unknown epitopes from the NY-ESO-1 tumor antigen. PMID- 15345305 TI - A novel ELISpot method for adherent cells. AB - The purpose of this study was to develop an enzyme-linked immunospot assay (ELISpot assay) that can be used with human adherent cells. While standard enzyme linked immunosorbent assays (ELISAs) are available and widely used and ELISpot assays are used for nonadherent lymphocytes, no ELISpot assay has been developed for adherent cells. We used primary human fibroblasts from four different tissues (myometrium, lung, gingiva, and orbit), either unstimulated or interleukin (IL) 1beta-activated, to evaluate an ELISpot assay. Antibody pairs for IL-6 and IL-8 were used and results were compared to a standard ELISA. We found that we could reliably detect IL-6 and IL-8 spots with as few as 10 fibroblasts. Optimal cell numbers were 50 cells per well incubated for 8 h, although spots appeared as early as 2 h after incubation. Spots were absent when cells, primary, or secondary anti-cytokine antibodies were omitted from the protocol. Spot number and size can be ascertained using current automated ELISpot reader technology. The frequency of IL-6 and IL-8-producing human fibroblasts could also be determined. For example, 60% of the lung fibroblasts express IL-6, but IL-8 can be detected from only 40% of the cells. Approximately 80% of the human orbital fibroblasts make IL-6, whereas approximately 50% generate IL-8 following IL-1beta stimulation. These new findings show that fibroblasts from different human tissues display different frequencies of cytokine production and this further supports the concept of fibroblast diversity. The sensitivity of this new ELISpot assay is adequate for cytokine detection in just a few cells, unlike the standard ELISA. It should permit ascertaining the frequency of fibroblasts and other adherent cells that produce cytokines and, if desired, can be used in tandem with a standard ELISA to determine total cytokine produced. Moreover, the assay is suitable for normal human adherent cells that are often short-lived and difficult to cultivate. PMID- 15345306 TI - Successful live cell harvest from bisected sentinel lymph nodes research report. AB - Sentinel lymph nodes provide an excellent opportunity to study early immune responses to cancer. However, harvesting live cells has not previously been possible, because it conflicts with the need to preserve tissue for histological interpretation. This study used scrape cytology on 26 sentinel and 8 non-sentinel nodes, harvested from 17 stage I/II melanoma patients undergoing sentinel node biopsy. Numbers of viable cells harvested before and after cryopreservation were measured and the effect on subsequent histology assessed. The mean number of cells harvested from 26 sentinel nodes was 7.06 x 10(6) (range 0.1-32.2), with a mean viability of 99.5% (range 87-100, lower 95% CI 98.5%). Furthermore, counts and viabilities were well maintained after cryopreservation. Flow cytometry confirmed CD3+, CD20+ and lineage-1-/HLA-DR+ subpopulations, consistent with T lymphocytes, B-lymphocytes and dendritic cells, respectively. Importantly, there was no discernible change in histological detail and the proportion of positive sentinel nodes remained unchanged. This technique will allow more functional and quantitative approaches to sentinel lymph node research. PMID- 15345307 TI - Responses of peptide-specific T cells to stimulation with polystyrene beads carrying HLA class I molecules loaded with single peptides. AB - Cell-sized microbeads carrying single peptide-loaded HLA class I molecules were prepared for HLA-A2 and HLA-B7 by a simple procedure which transfers single peptide-loaded HLA class I molecules from cultured cells to polystyrene beads using anti-peptide antibodies directed to an intracellular segment of HLA-A alpha chains. The surface density of peptide-loaded HLA class I molecules on beads was comparable to that on the peptide-loaded cells. HLA-A2 beads loaded with an HCV peptide HCV1073 were tested for stimulation activity on an HCV1073-specific CD8+ T cell clone NS3-1. A substantial level of gamma-IFN production was induced. The stimulation was peptide-specific. The efficiency was dependent on the bead concentration and the surface HLA class I density on beads and enhanced significantly by co-coupling of anti-CD28 to peptide-loaded beads. The peptide loading efficiency on HLA class I molecules and the transfer efficiency of HLA class I molecules to polystyrene beads were reasonably high for HLA-A2 and HLA B7. Thus, polystyrene beads carrying these single peptide-loaded HLA class I molecules are potentially useful in further analysis of the co-stimulatory or inhibitory factors involved in CD8+ T cell responses and eventually in detection of cytotoxic T cells in PBLs. PMID- 15345308 TI - Development of a monoclonal antibody-based enzyme-linked immunoabsorbent assay for the binding of gp130 to the IL-6/IL-6R complex and its competitive inhibition. AB - The proinflammatory cytokine IL-6 binds to the membrane bound or soluble IL-6 receptor (IL-6R) and activates an intracellular signaling cascade after complex formation with two gp130 molecules. These mediate general homeostasis and orchestrates the immune response during disease. Trans-signalling via the soluble IL-6R has importance for the development and maintenance of human diseases like Crohn's disease, peritonitis and rheumatoid arthritis. We have developed an enzyme-linked immunoabsorbent assay (ELISA) that detects the binding of gp130 to the IL-6/sIL-6R complex. Competitive binding of sgp130-Fc, viral IL-6, and the inhibitory drug Suramin to gp130 has been demonstrated. The assay can be used for high-throughput screening of peptide and chemical compound libraries for the identification of new agonists and antagonists of the binding between gp130 and IL-6/sIL-6R. PMID- 15345309 TI - The induction of eosinophil peroxidase release: improved methods of measurement and stimulation. AB - The production and release of eosinophil peroxidase (EPO) has been associated with human pathology. Degranulation assays with eosinophils are typically very difficult to do, with very low release values. EPO is unique for its high cationic charge. As such, it adheres to most extracellular surfaces, rendering it more difficult to measure compared with other released cellular proteins. Based on the understanding of the sticky nature of EPO, we were concerned that EPO released in vitro cannot be reproducibly measured in the supernatants of stimulated cells. Instead, we suspected that much of the released EPO was left adherent to the tube walls. We chose to investigate the measurement of EPO activity using the peroxidase substrate, O-phenylenediamine (OPD). Unlike other peroxidase substrates, OPD is soluble in aqueous physiological solutions, which do not lyse cell membranes, thereby allowing us to add OPD directly to eosinophils and exclusively measure extracellular EPO. This novel approach would remove the concerns of incorrect EPO measurements due to its adhesive nature. In addition, we developed this method to quantify EPO release in terms of EPO concentration. Finally, using this technique, we have been able to demonstrate secretory IgA (s-IgA)-induced release of EPO. By using OPD, we have developed a more sensitive and specific method to analyze the release of extracellular EPO. PMID- 15345310 TI - High efficiency creation of human monoclonal antibody-producing hybridomas. AB - The native human antibody repertoire holds unexplored potential for the development of novel monoclonal antibody therapeutics. Current techniques that fuse immortal cells and primary B-lymphocytes are sub-optimal for the routine production of hybridomas that secrete human monoclonal antibodies. We have found that a murine cell line that ectopically expresses murine interleukin-6 (mIL-6) and human telomerase (hTERT) efficiently forms stable human antibody-secreting heterohybridomas through cell fusion with primary human B-lymphocytes. The hybrid cells maintain secretion of human antibodies derived from the primary B lymphocytes through multiple rounds of cloning. Using splenic B-lymphocytes from a patient immunized with a Streptococcus pneumoniae capsular polysaccharide vaccine, we have succeeded in creating hybridomas that secrete human monoclonal antibodies specific for S. pneumoniae antigens. Using peripheral blood lymphocytes, we have similarly cloned a human antibody that binds a viral antigen. These experiments establish that SP2/0-derived cell lines ectopically expressing mIL-6 and hTERT will enable the rapid cloning of native human monoclonal antibodies. PMID- 15345311 TI - A cell-based time-resolved fluorescence assay for selection of antibody reagents for G protein-coupled receptor immunohistochemistry. AB - A cell-based time-resolved fluorescence (celTRF) immunoassay is described for pre screening antibodies to G protein-coupled receptor (GPCR) peptides that predicts suitability for immunohistochemistry (IHC). Rat GPCRs were expressed in Saos-2 human osteosarcoma cells via recombinant baculoviruses designed for mammalian cell expression, i.e., the transduced cells were used as a "screening lawn". The lawn was fixed and permeabilized similarly to IHC tissue. The celTRF, a dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA), employed Eu labelled goat anti-rabbit IgG. It exhibited a broad dynamic range upon which enzyme-linked immunosorbant assay (ELISA)-positive affinity-purified anti-peptide antibody reagents were examined for specificity and potency. Over 150 anti peptide reagents to 27 GPCRs were characterized. All celTRF-positive antibodies were found to be suitable for IHC, whereas ELISA alone did not predict IHC utility. Examples are illustrated with five rabbit anti-neuropeptide FF receptor 1 (NPFF1) antibodies, where a strong correlation between celTRF potency and IHC utility was observed in both applications. In contrast, two high anti-peptide ELISA titer but celTRF-negative antibodies failed to recognize the NPFF1 receptor in IHC. The celTRF assay was performed manually and in an automated fashion, in our case, using a Biomek FX station and Sami scheduling software. The celTRF is the first in vitro automated assay that offers confident pre-selection of antibodies for IHC and the versatility to accommodate the rapid screening of large numbers of GPCRs. The celTRF is readily applicable to other protein target classes. PMID- 15345312 TI - Isolation of human prostate cancer cell reactive antibodies using phage display technology. AB - Here we describe a phage display strategy for the selection of rabbit monoclonal antibodies that recognize cell surface tumor-associated antigens expressed in prostate cancer. Two immune rabbit/human chimeric Fab libraries were displayed on phage and used to search for tumor-associated antigens by panning on DU145 human prostate cancer cells. For this, we developed a novel whole-cell panning protocol with two negative selection steps designed to remove antibodies reacting with common antigens. After three rounds of subtractive panning, a majority of clones bound to DU145 cells as detected by flow cytometry. Among these, we identified several clones that bound selectively to DU145 cells but not to primary human prostate epithelial cell line PrEC. In summary, our work demonstrates the potential of immune rabbit antibody libraries for target discovery in general and the identification of cell surface tumor-associated antigens in particular. PMID- 15345313 TI - Use of retroviral vectors for the analysis of SIV/HIV-specific CD8 T cell responses. AB - CD8+ T cell responses and particularly cytotoxic T lymphocyte (CTL) activity are critical factors in controlling SHIV, SIV or HIV replication during natural infection and represent key parameters which need to be monitored during vaccine development. In order to improve the methodology for measuring CD8+ T cell responses, retroviral vectors expressing the full-length SIV-Gag or HIV-Env proteins were constructed and used to transduce B lymphoblastoid cell lines (BLCL) from cynomolgus monkeys infected with SHIV89.6P. Continuous expression of Gag and Env proteins was detected in stably transduced BLCL, which induced Gag- or Env-specific T cell responses, as measured by both IFNgamma-ELISPOT and chromium release assays, upon in vitro stimulation of PBMC from the SHIV89.6P infected monkeys. Moreover, induction of Gag-specific CTL using BLCL transduced with retroviral vector expressing the SIV-Gag protein was more efficient and specific compared to that obtained using BLCL infected with a recombinant vaccinia virus (rVV) encoding for the same antigen. Assays on purified CD4+ and CD8+ T cells indicated that both populations specifically produced IFNgamma, but only the CD8+ T cells mediated Gag- and Env-specific cytotoxicity, indicating preferential expansion of these effector cells. Thus, this method represents an alternative tool for the analysis of CTL responses during vaccination protocols in those animal models where little information is available on MHC class I alleles or CTL epitopes. PMID- 15345314 TI - Chemiluminescent immunosensor for CA19-9 based on antigen immobilization on a cross-linked chitosan membrane. AB - A novel chemiluminescent immunosensor for carbohydrate antigen 19-9 (CA19-9) based on the immobilization of CA19-9 on the cross-linked chitosan membrane was developed. The different membranes were characterized by atomic force microscopy (AFM) and infrared spectrum, respectively. Based on a noncompetitive immunoassay format, this proposed chemiluminescent immunosensor enabled a low-cost, flexible and rapid determination for CA19-9 in combination with flow injection analysis (FIA). After an off-line incubation of the analyte CA19-9 with horseradish peroxidase (HRP)-labeled anti-CA19-9, the mixture was injected into the immunosensor, which led to the trapping of free HRP-labeled anti-CA19-9 by the immobilized antigen in the immunosensor. The trapped HRP-labeled antibody was detected by chemiluminescence due to its catalytic activity following the reaction of luminol and H2O2. Under optimal conditions, the decreased chemiluminescent signal of the immunosensor was proportional to the CA19-9 concentration in the range of 2.0-25 U/ml with a detection limit of 1.0 U/ml. The immunosensor showed an acceptable accuracy and good reproducibility. The results of 20 human serum samples detected by this method were in acceptable agreement with those obtained by immunoradiometric assay. The proposed immunosensor provided a new promising tool for practical clinical detection of the serum CA19 9 level. PMID- 15345315 TI - Evaluation of Elispot assays: influence of method and operator on variability of results. AB - In this study, a comprehensive comparative analysis of different evaluation methods of Elispot plates was performed. Three investigators using three different evaluation approaches read 50 randomly selected wells at three different time points. The methods were: (1) manual evaluation using a stereomicroscope, (2) automated evaluation using an image analysis reader system with reading parameters established by each investigator, and (3) automated evaluation using a reader system with preset reading parameters using assay specific controls. We demonstrate that manual evaluation had the highest variability both within the same method and when comparing all methods, followed by automated evaluation with investigator-dependent parameters. The variability was low only when all investigators used the same parameters established using assay-specific controls. This variability was independent of operator or spot number per well. Based on this study, recommendations for standardization and validation procedures of Elispot assay performance and evaluation procedures are presented. PMID- 15345316 TI - Novel application of a whole blood intracellular cytokine detection assay to quantitate specific T-cell frequency in field studies. AB - We optimized a whole blood intracellular cytokine assay to quantitate the frequency of specific CD4+ and CD8+ T cells in small volumes of whole blood from infants from developing countries. The assay is performed in two steps. First, whole blood is stimulated in the presence of specific antigens for 6-18 h, ending with cryopreservation of fixed white cells. These stimulation steps were specifically adapted to be practical and reliable in a rural, developing country field setting. Later, in a more resourceful setting, interferon-gamma producing CD4+ or CD8+ T cells are detected by flow cytometry. The assay proved sensitive and specific for detecting mycobacteria-specific immunity 10 weeks after Bacillus Calmette-Guerin (BCG) vaccination of newborns from a rural field site. PMID- 15345317 TI - The novel histone deacetylase inhibitor BL1521 inhibits proliferation and induces apoptosis in neuroblastoma cells. AB - Neuroblastoma is a childhood cancer arising from the sympathetic nervous system. Disseminated neuroblastoma has a poor prognosis despite intensive multimodality treatment. Histone deacetylases (HDACs) were recently discovered as a potential target for pharmacological gene therapy in cancer. HDACs have an important function in regulating DNA packaging in chromatin, thereby affecting the transcription of genes. In this paper, we tested the efficacy of a newly developed histone deacetylase inhibitor, BL1521, on neuroblastoma in vitro by investigating the changes in: acetylation of histone H3, in situ HDAC activity, p21(WAF1/CIP1) and MYCN expression, metabolic activity, proliferation, morphology and the amount of apoptosis present. BL1521 inhibited the in situ HDAC activity of a panel of neuroblastoma cell lines by at least 85%. Western analysis showed an increase of histone H3 acetylation in neuroblastoma cells after incubation with BL1521. Northern analysis showed an increase in the expression of p21(WAF1/CIP1) and a decrease in the expression of MYCN in neuroblastoma cells after incubation with BL1521. Proliferation as well as the metabolic activity of neuroblastoma cells decreased significantly in response to treatment with BL1521, regardless of the MYCN status of the cells. BL1521 induced poly-(ADP-ribose) polymerase cleavage in a time- and dose-dependent manner, indicating the induction of apoptosis. Furthermore, when compared to the HDAC inhibitors Trichostatin A and 4-phenylbutyrate, BL1521 has an intermediate efficacy. Our results show that BL1521 is a potent inhibitor of HDAC and that HDACs are an attractive target for selective chemotherapy in neuroblastoma. PMID- 15345318 TI - Tissue distribution and activity studies of 1,24-dihydroxyvitamin D2, a metabolite of vitamin D2 with low calcemic activity in vivo. AB - The active vitamin D compound 1alpha,24(S)-dihydroxyvitamin D(2) (1,24(OH)(2)D(2)) is under development as a therapy for disorders including cancer and secondary hyperparathyroidism. 1,24(OH)(2)D(2) is a potent inhibitor of cell proliferation in vitro and, relative to calcitriol (1,25(OH)(2)D(3)), has reduced calcemic activity in vivo. To examine the mechanisms underlying this reduced calcemic activity, we studied the tissue distribution in rats of radiolabeled 1,24(OH)(2)D(2) or 1,25(OH)(2)D(3) over 24h. Serum levels of 1,24(OH)(2)D(2) were lower than those of 1,25(OH)(2)D(3) at all time points; however, tissue levels of radiolabeled compounds followed different patterns. In duodenum and kidney, 1,24(OH)(2)D(2) and 1,25(OH)(2)D(3) rose to similar levels at early time points; 1,24(OH)(2)D(2) levels then declined more rapidly. In bone marrow, 1,24(OH)(2)D(2) and 1,25(OH)(2)D(3) were present at similar levels at all time points. In liver, 1,24(OH)(2)D(2) levels were two-fold higher than 1,25(OH)(2)D(3) at 1h post-injection, declining to similar levels by 8h. In vitamin D-deficient rats, doses of 1,24(OH)(2)D(2) 30-fold higher than 1,25(OH)(2)D(3) were required to produce equal stimulation of intestinal calcium absorption. In the same deficient animals, 1,24(OH)(2)D(2) and 1,25(OH)(2)D(3) were nearly equipotent at stimulating bone calcium mobilization. In cultured bone cells, 1,24(OH)(2)D(2) and 1,25(OH)(2)D(3) were equipotent at stimulating osteoclast formation and bone resorption. In summary, the reduced calcemic activity of 1,24(OH)(2)D(2) may result from altered pharmacokinetics relative to 1,25(OH)(2)D(3), resulting in relatively rapid decreases in 1,24(OH)(2)D(2) levels and activity in target organs such as intestine. Further studies will be necessary to confirm these findings and to confirm the clinical utility of 1,24(OH)(2)D(2). PMID- 15345319 TI - Cyclic antimicrobial peptides based on Limulus anti-lipopolysaccharide factor for neutralization of lipopolysaccharide. AB - Bacterial endotoxin (lipopolysaccharide, LPS) is responsible for the septic shock syndrome. As potential therapeutic agents cyclic cationic antimicrobial peptides of different length, based on the Limulus anti-lipopolysaccharide factor (LALF), were synthesized, and their interaction with LPS was characterized physico chemically and related to results in biological assays. All peptides inhibited the LPS-induced cytokine production in human mononuclear cells and the Limulus amebocyte lysate in a concentration-dependent way, with the peptide comprising the complete LPS-binding loop of the LALF (cLALF22) being the most effective. The peptides were neither cytotoxic nor hemolytic, except a slight effect of cLALF22. The peptides were able to displace Ca(2+) cations from a LPS monolayer, with cLALF22 being again most effective in accordance with results from isothermal titration calorimetry, in which saturation of binding was observed at an equimolar [cLALF22]:[LPS] ratio, and at a ratio 2-2.5 for the other peptides. For cLALF22, zeta (xi) potential experiments exhibited a complete compensation of the negative charges of LPS, whereas for the other peptides a residual negative potential of -20 to -40mV was found. X-ray diffraction experiments showed that the mixed unilamellar/cubic inverted aggregate structure of the lipid A part of LPS was converted into a multilamellar one. The gel to liquid crystalline phase transition of the acyl chains of LPS was changed upon cLALF22 binding, leading to a clear fluidization, which was not observed or only to a lesser degree for the other peptides. The affinity of the peptides for LPS led to a reduced binding of lipopolysaccharide-binding protein (LBP) to target membranes and hence to an inhibition of cytokine induction in human mononuclear cells. PMID- 15345320 TI - Insulin and glucose induced changes in expression level of nucleoside transporters and adenosine transport in rat T lymphocytes. AB - Adenosine is an endogenous agent exerting potent action on the immune system including regulation of lymphocyte functioning. Impaired T lymphocyte functioning is a common feature of diabetes. The aims of this study were to examine the effects of glucose and insulin on nucleoside transporters (NT) expression level and adenosine (Ado) transport in rat T lymphocytes cultured under the defined concentrations of glucose and insulin. Performed experiments revealed that rat T lymphocytes expressed the equilibrative nucleoside transporter type 1 and 2 (rENT1, rENT2) and concentrative nucleoside transporter type 2 (rCNT2). The mRNA levels of rENT2 and rCNT2 were highly dependent on insulin but were not affected by changes in extracellular glucose concentration. Exposition of T cells to 10nM insulin resulted in 73% increase in rENT2 mRNA and 50% decrease in the rCNT2 mRNA level. The level of rENT1 mRNA was sensitive to extracellular glucose concentration but not to insulin. The highest differences among cells cultured in high (20mM) and low (5mM) glucose were observed in equilibrative nitrobenzylthioinosine sensitive adenosine transport, which was lowered by 65% in cells cultured at high glucose. Alterations in adenosine transport were accompanied by changes in adenosine accumulation in the cell. These results indicate that adenosine transport in rat T lymphocytes is independently and differentially regulated by glucose and insulin by means of changes in the nucleoside transporters expression level. Altered adenosine transport has a great impact on its intracellular level. This suggests that under diabetic conditions adenosine action on T lymphocytes might be altered. PMID- 15345321 TI - Immunomodulatory effects of thalidomide analogs on LPS-induced plasma and hepatic cytokines in the rat. AB - Thalidomide has shown to inhibit, selectively and mainly the cytokine tumor necrosis factor-alpha (TNF-alpha), thus, thalidomide has inhibitory consequences on other cytokines; this is ascribed as an immunomodulatory effect. Novel thalidomide analogs are reported with immunomodulatory activity. The aim of this work was to synthesize some of these analogs and to assess them as immunomodulatory agents in an acute model of LPS-induced septic challenge in rat. Animal groups received orally twice a day vehicle carboxymethylcellulose (0.9%), or thalidomide in suspension (100mg/kg), or analogs in an equimolar dose. Two hours after last dose, rats were injected with saline (NaCl, 0.9%, i.p.) or LPS (5mg/kg, i.p.). Groups were sacrificed 2h after injection and samples of blood and liver were obtained. TNF-alpha, interleukin-6, -1beta, and -10 (IL-6, IL 1beta, IL-10) were quantified by enzyme linked immunosorbent assay (ELISA) and studied in plasma and liver. After 2h of LPS-induction, different patterns of measured cytokines were observed with thalidomide analogs administration evidencing their immunomodulatory effects. Interestingly, some analogs decreased significantly plasma and hepatic levels of LPS-induced proinflammatory TNF-alpha and others increased plasma concentration of anti-inflammatory IL-10. Thalidomide analogs also showed slight effects on the remaining proinflammatory cytokines. Differences among immunomodulatory effects of analogs can be related to potency, mechanism of action, and half lives. Thalidomide analogs could be used as a pharmacological tool and in therapeutics in the future. PMID- 15345322 TI - Callipeltin A: sodium ionophore effect and tension development in vascular smooth muscle. AB - Callipeltin A is a cyclic depsidecapeptide isolated from the marine sponges Callipelta sp. and Latrunculia sp. that has been previously shown to increase the force of contraction of guinea-pig atria through the inhibition of Na+/Ca2+ exchanger (NCX). We investigated the effect of callipeltin A on guinea-pig aortic rings contracted by procedures that activate NCX in "calcium entry mode". Callipeltin A did not inhibit these contractions. Resting aorta responded to callipeltin A with a remarkable contraction that was concentration-dependent (EC50 0.44microM). This contraction was not inhibited by the calcium channel blocker verapamil and was not mediated by the activation of alpha-adrenergic or endothelin-1 receptors. Pre-incubation of aortic rings with 0.5mM amiloride, an inhibitor of NCX, completely prevented callipeltin A-induced contraction. Furthermore, callipeltin A (EC50 0.51microM) increased Na+ efflux of Na-loaded erythrocytes. 1H and 13C NMR resonances of callipeltin A revealed small but significant changes in the titration with K+ and Na+ salts. It is suggested that the effect of callipeltin A on cardiac and vascular preparations is linked to a Na-ionophore action. PMID- 15345323 TI - UCP3 and thyroid hormone involvement in methamphetamine-induced hyperthermia. AB - Here, we determined the extent of hypothalamic-pituitary-thyroid (HPT) axis and uncoupling protein-3 (UCP3) involvement in methamphetamine (METH)-induced hyperthermia. Sprague-Dawley rats treated with METH (40mg/kg, s.c.) responded with a hyperthermic response that peaked 1h post-treatment and was sustained through 2h. After METH treatment, thyroparathyroidectomized (TX) animals developed hypothermia that was sustained for the 3h monitoring period. In TX animals supplemented for 5 days with levothyroxine (100microg/kg, s.c.), METH induced hypothermia was eliminated and the hyperthermic response was restored. Thyroid hormone levels (T3 and T4), measured in euthyroid animals 1h after METH, remained unchanged. As seen in rats, 1h post-METH (20mg/kg, i.p.) treatment, wild type (WT) mice developed profound hyperthermia that was sustained for 2h. In marked contrast, UCP3-/- animals developed a markedly blunted hyperthermic response at 1h compared to WT animals. Furthermore, UCP3-/- mice could not sustain this slight elevation in temperature. Two hours post-METH treatment, UCP3 /- animal temperature returned to baseline temperatures. UCP3-/- mice were also completely protected against the lethal effects of METH, whereas 40% of WT mice succumbed to the hyperthermia. These findings suggest that thyroid hormone plays a permissive role in the thermogenic effects induced by METH. Furthermore, the findings indicate that UCP3 plays a major role in the development and maintenance of the hyperthermia induced by METH. The relationship of these results to the hyperthermia induced by 3,4-methylenedioxymethamphetamine (MDMA) is also discussed. PMID- 15345324 TI - Mitochondria-mediated apoptosis operating irrespective of multidrug resistance in breast cancer cells by the anticancer agent prodigiosin. AB - Prodigiosin (PG) is a red pigment produced by Serratia marcescens with pro apoptotic activity in haematopoietic and gastrointestinal cancer cell lines, but no marked toxicity in non-malignant cells. Breast cancer is the most frequent malignancy among women in the European Union and better therapies are needed, especially for metastatic tumors. Moreover, multidrug resistance is a common phenomenon that appears during chemotherapy, necessitating more aggressive treatment as prognosis worsens. In this work, we extend our experiments on PG induced apoptosis to breast cancer cells. PG was potently cytotoxic in both estrogen receptor positive (MCF-7) and negative (MDA-MB-231) breast cancer cell lines. Cytochrome c release, activation of caspases-9, -8 and -7 and cleavage of poly (ADP-ribose) polymerase protein typified the apoptotic event and caspase inhibition revealed that PG acts via the mitochondrial pathway. In a multidrug resistant subline of MCF-7 cells that over-expresses the breast cancer resistance protein, the cytotoxic activity of PG was slightly reduced. However, flow cytometry analysis of PG accumulation and efflux in MCF-7 sublines showed that PG is not a substrate for this resistance protein. These results suggest that PG is an interesting and potent new pro-apoptotic agent for the treatment of breast cancer even when multidrug resistance transporter molecules are present. PMID- 15345325 TI - Evidence for multiple glucuronide transporters in rat liver microsomes. AB - The transport of glucuronides across the endoplasmic reticulum membrane is an important step in the overall process of biotransformation, although the mechanism remains unclear and the participating transporters are unidentified. Using a rapid filtration assay in combination with liquid chromatography-mass spectrometry, we measured the transport of a variety of beta-D-glucuronides in rat liver microsomes and investigated the substrate specificity of the participating transporter(s) by inhibition studies. Time-dependent and bi directional transport of phenolphthalein glucuronide was detected and the kinetic parameters for transport were determined. The K(m) and V(max) values of high affinity transport were 26microM and 3.9nmol/min/mg protein, respectively. Phenolphthalein glucuronide transport was inhibited by 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid and N-ethylmaleimide. Transport inhibition studies revealed competition between three glucuronides: phenolphthalein glucuronide, estradiol 17-glucuronide and naphthol AS-BI glucuronide indicating that they share a common transporter in the endoplasmic reticulum membrane. Their transport was inhibited by phenolphthalein, but was not affected by p-nitrophenyl glucuronide, naphthyl glucuronide or d-glucuronate. Morphine 3-glucuronide transport was not inhibited by any of the latter four compounds or by phenolphthalein glucuronide. This novel experimental approach has produced data consistent with the presence of multiple (at least three) transporters catalyzing the transport of glucuronides through the endoplasmic reticulum membrane. These data also indicate that the size and/or shape of the aglycone rather than the glucuronic acid moiety per se is an important determinant of transporter specificity. PMID- 15345326 TI - Induction of cellular resistance to nucleoside reverse transcriptase inhibitors by the wild-type breast cancer resistance protein. AB - Breast cancer resistance protein (BCRP/ABCG2) is a novel member of ATP-binding cassette transporters, which induce multidrug resistance in cancer cells. We previously reported that a high level of BCRP expression in CD4(+) T cells conferred cellular resistance to nucleoside reverse transcriptase inhibitors (NRTIs) of human immunodeficiency virus type 1 (HIV-1). However, this BCRP was found to have a mutation of Arg to Met at position 482 (BCRP(R482M)). The present study demonstrated that the wild-type BCRP (BCRP(WT)) also conferred cellular resistance to NRTIs. MT-4 cells (a CD4(+) T-cell line) highly expressing BCRP(WT) (MT-4/BCRP) were generated and the expression of BCRP(WT) was confirmed by genotypic and phenotypic analyses. Compared to the parental MT-4 cells, MT-4/BCRP cells displayed resistance to zidovudine (AZT) in terms of antiviral activity as well as drug cytotoxicity. In addition, other NRTIs were also less inhibitory to HIV-1 replication in MT-4/BCRP cells than in MT-4 cells. Significant reduction of intracellular AZT accumulation was observed in MT-4/BCRP cells. An analysis for intracellular metabolism of AZT suggested that the resistance was attributed to the increased efflux of AZT and its metabolites in MT-4/BCRP cells. Furthermore, the BCRP-specific inhibitor fumitremorgin C completely restored the reduction of AZT in MT-4/BCRP cells. These results indicate that, like BCRP(R482M), BCRP(WT) also plays an important role in cellular resistance to NRTIs. PMID- 15345327 TI - Determination of interleukin-4-responsive region in the human cytochrome P450 2E1 gene promoter. AB - Cytochrome P450 2E1 (CYP2E1) gene expression is known to be induced by interleukin-4 (IL4) and repressed by inflammatory cytokines, such as interleukin 1beta3 (IL1beta3) in human hepatocytes. The mechanisms involved in these transcriptional regulations remain elusive. In order to study these mechanisms, various constructs of the human CYP2E1 promoter were prepared and transfected into the human HepG2 hepatoma cell line. Our findings revealed that an IL4 responsive region of 128bp (-671/-544) was required to mediate induction by IL4. IL1beta caused moderate but significant decrease of the promoter activity, which was abolished when the two cytokines were combined. The IL1beta inhibitory effect is mediated through a regulatory sequence independent of that of IL4. Furthermore, by using specific signaling pathway inhibitors, we demonstrated that IL4 activation required protein kinase C (PKC) activation. In addition, our results suggest that induction by IL4 was not dependent on a single binding site but rather on a complex region which includes putative binding sites for signal transducer and activator of transcription (STAT)6, activator protein (AP)-1, nuclear factor kappa-B (NFkappaB), nuclear factor of activated T cells (NFAT) and CCAAT enhancer binding protein (C/EBP). Electrophoretic mobility shift assays suggest that AP1 and NFAT transcription factors are able to bind to three sites in the IL4-responsive region. PMID- 15345328 TI - GABA(A)/central benzodiazepine receptor and peripheral benzodiazepine receptor ligands as inducers of phenobarbital-inducible CYP2B and CYP3A. AB - A sequence critical for phenobarbital (PB) induction, the PB response unit (PBRU), situated upstream of the rat CYP2B1 and CYP2B2 genes, includes two nuclear receptor binding sites, NR1 and NR2. When NR1 and NR2 are mutated PB responsiveness is abolished. While no nuclear receptor for which PB is an agonist ligand has yet been identified, PB is a ligand of GABA(A) receptors and it can displace [(3)H] 1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3 isoquinolinecarboxamide (PK 11195) from its binding site on the peripheral benzodiazepine receptor (PBR). We assessed CYP2B levels in primary rat hepatocytes following treatment with 10 ligands of either or both of these receptors. All compounds tested were found to be CYP2B1/CYP2B2 inducers and most were CYP3A inducers. Five had not previously been described as CYP2B1/CYP2B2 inducers: bicuculline, flunitrazepam, 4'-chlorodiazepam (Ro5-4864), N,N-dihexyl-2 (4-fluorophenyl)indole-3-acetamide (FGIN 1-27) and 7-(dimethylcarbamoyloxy)-6 phenylpyrrolo-[2,1-d][1,5]benzothiazepine (DCPPBT). Reporter gene analysis demonstrated that CYP2B induction by these agents and other PBR or GABA(A) receptor ligands is mediated through the PBRU and the NR1/NR2 sites, suggesting a molecular mechanism similar to that for PB induction. The potencies for PBRU dependent induction by 11 ligands of PBR or the GABA(A) receptor was evaluated. FGIN-127, DCPPBT and PK 11195 exhibited EC(50) values for PBRU-dependent transcription activation about three orders of magnitude higher than the reported affinities of the PBR for these agents, arguing against the involvement of the PBR in PB induction. However the EC(50) values found for the agents tested encourage further investigation on the possible involvement of the GABA(A) receptor in PB induction. PMID- 15345329 TI - Nephroblastoma overexpressed gene (NOV) expression in rat hepatic stellate cells. AB - Using the expression-profiling method, we identified nephroblastoma overexpressed gene (NOV) mRNA as one member of the mRNA population that was upregulated in cultured activated hepatic stellate cell (HSC). Northern analysis showed that NOV mRNA was increasingly expressed during progressive activation of cultured rat HSCs, and a significant increase was observed in both the carbon tetrachloride induced and bile duct ligation/scission rat models of liver fibrosis. RT-PCR showed human NOV mRNA was increased in most fibrotic livers compared with normal livers. The expression of NOV protein in fibrotic rat and human livers was predominantly located in areas of ductular proliferation and HSC of the fibrous septa. HSCs stimulated with transforming growth factor beta1 showed increased expression of NOV protein without changing its mRNA levels. Dexamethasone stimulated the expression of NOV mRNA and protein. Furthermore, we demonstrated that bile acids have a modulating effect on the induction of NOV mRNA expression. In conclusion, this study suggests that NOV is expressed during liver fibrogenesis and HSCs may be an important source of hepatic NOV. PMID- 15345330 TI - Inhibition of hyaluronan export from human fibroblasts by inhibitors of multidrug resistance transporters. AB - In a previous report we described the export of hyaluronan from Streptococcus pyogenes by an ABC transporter. Extending these findings a sequence homology search against human proteins revealed a strong homology to the multidrug resistance transporter ABC-B (MDR-1) and ABC-C (MRP 5). Using several inhibitors directed against these and other transporters, a decreased hyaluronan production in cell culture as well as in hyaluronan synthase activity in purified membrane fractions was observed. The inhibitory capacity (IC(50) concentrations) was compared the with reported IC(50)- or the K(i)-concentrations for individual transporters. These analyses revealed that hyaluronan is synthesized within the cytoplasm of mammalian cells and actively secreted into the pericellular space by energy dependent transport proteins. While inhibition of several transport proteins resulted in a decrease of hyaluronan export, inhibition of the MRP5 transporter was the most effective one to decrease hyaluronan in the cell culture supernatant indicating that hyaluronan export is one physiological role of this transport protein. PMID- 15345331 TI - Prevention of age-related changes in rat cortex transcription factor activator protein-1 by hypolipidemic drugs. AB - We sought to investigate if, similar to what has been described in other rodent tissues, ageing changes the activity of several transcription factors, namely signal transducer and activator of transcription, nuclear factor-kappa B (NFkappaB), activated protein-1 (AP-1) and peroxisome proliferator-activated receptor (PPAR), in cortex of Sprague-Dawley rats. We also investigated if the administration of two hypolipidemic drugs, gemfibrozil (GFB) and atorvastatin (ATV), could prevent those changes. To this purpose, we determined the expression and binding activity of these transcription factors in cortex samples from 3 month and 18-month old male and female rats, and in 18-month old rats of both sexes treated for 21 days with a daily dose of 3mg GFB/kg or 10mg ATV/kg. Ageing increased rat cortex NFkappaB binding activity by 35-40%, and decreased by 22-26% the amount of PPARalpha in rats of both sexes, while cortex AP-1 binding activity and c-Jun content were reduced only in old females (-26 and -50%, respectively). Cortex cyclooxigenase-2 (COX-2) and receptor for activated C-kinase 1 (RACK1) expression was also reduced by old age. Hypolipidemic drugs prevented the age related decrease of cortex AP-1 in old females and increased AP-1 binding activity and c-Jun protein in cortex from both old male and female rats. GFB increased also by 80% the cortex PPARalpha content in old males. Our data indicate that 18-month old rats show signs of cortex biochemical deterioration related to the ageing process, and that hypolipidemic drug administration partially prevents the appearance of some of the age-related changes in cortex biochemistry. PMID- 15345332 TI - Activation of osteoblastic functions by a mediator of pain, bradykinin. AB - We investigated the effects of bradykinin (BK) on the production of interleukin (IL)-6 and prostaglandin PGE(2), whose molecules are capable of stimulating the development of osteoclasts from their hematopoietic precursors as well as the signal transduction systems involved, in human osteoblasts (SaM-1 cells). BK receptors B1 (B1R) and B2 (B2R) were expressed in SaM-1 and osteosarcoma (SaOS-2, HOS, and MG-63) cells. Treatment of SaM-1 cells with BK increased the synthesis of both IL-6 and PGE(2) and the increase in both was blocked by HOE140 (B2R antagonist), but not by Des-Arg(9)-[Leu(8)]-BK (B1R antagonist). U-73122, a phospholipase C (PLC) inhibitor, suppressed BK-induced IL-6 and PGE(2) synthesis in SaM-1 cells. In addition, BK caused an increase in the intracellular Ca(2+) concentration ([Ca(2+)]i), which was inhibited by pretreatment with HOE140 or 2 aminoethoxydiphenyl borate (2-APB), an inositol 1,4,5-trisphosphate (IP(3)) receptor (IP(3)R) blocker. Furthermore, both SB203580 (an inhibitor of p38 mitogen-activated protein kinase [MAPK]) and PD98059 (an inhibitor of MEK, upstream of ERK) attenuated the BK-induced IL-6 and PGE(2) synthesis. BK treatment resulted in the phosphorylation of p38 MAPK and extracellular signal regulated kinase (ERK)1/2, and 2-APB could suppress BK-induced phosphorylation of ERK1/2. These findings suggest that BK increased both IL-6 and PGE(2) synthesis in osteoblastic cells via B2R and that PLC, IP(3)-induced [Ca(2+)]i, MEK, and MAPKs were involved in the signal transduction in these cells. PMID- 15345333 TI - The cytotoxic mechanism of glyoxal involves oxidative stress. AB - Glyoxal is a reactive alpha-oxoaldehyde that is a physiological metabolite formed by lipid peroxidation, ascorbate autoxidation, oxidative degradation of glucose and degradation of glycated proteins. Glyoxal is capable of inducing cellular damage, like methylglyoxal (MG), but may also accelerate the rate of glycation leading to the formation of advanced glycation end-products (AGEs). However, the mechanism of glyoxal cytotoxicity has not been precisely defined. In this study we have focused on the cytotoxic effects of glyoxal and its ability to overcome cellular resistance to oxidative stress. Isolated rat hepatocytes were incubated with different concentrations of glyoxal. Glyoxal by itself was cytotoxic at 5mM, depleted GSH, formed reactive oxygen species (ROS) and collapsed the mitochondrial membrane potential. Glyoxal also induced lipid peroxidation and formaldehyde formation. Glycolytic substrates, e.g. fructose, sorbitol and xylitol inhibited glyoxal-induced cytotoxicity and prevented the decrease in mitochondrial membrane potential suggesting that mitochondrial toxicity contributed to the cytotoxic mechanism. Glyoxal cytotoxicity was prevented by the glyoxal traps d-penicillamine or aminoguanidine or ROS scavengers were also cytoprotective even when added some time after glyoxal suggesting that oxidative stress contributed to the glyoxal cytotoxic mechanism. PMID- 15345334 TI - Dynamics of interactions of photosensitizers with lipoproteins and membrane models: correlation with cellular incorporation and subcellular distribution. AB - The incorporation and subcellular localization of photosensitizers are critical determinants of their efficiency. Here, we correlate these properties with the interactions of photosensitizers with membrane-models and low density lipoproteins (LDL) in acellular systems. Focus was given on dynamics aspects. Two amphiphilic photosensitizers, deuteroporphyrin (DP) and aluminum phthalocyanine sulfonated on two adjacent isoindole units (AlPcS2a) were selected. The phthalocyanine was bound to LDL with an overall association constant around 5 x 10(7)M(-1). Biphasic association kinetics was indicative of two types of sites. The release of the phthalocyanine into the bulk aqueous medium occurred within less than a second. A similar behavior was found previously for deuteroporphyrin although its affinity was somewhat higher (5.5 x 10(8)M(-1)). Both compounds were previously characterized by high affinity for membrane-models and quick exchange with the bulk solution. However, they strongly differed by their rate of transfer through the lipid bilayer, in the range of seconds for the porphyrin, several hours for the phthalocyanine. In the case of the porphyrin, fluorescence microscopy on human fibroblasts showed diffuse labeling with no significant modification of the distribution upon vectorization by LDL. In contrast, the phthalocyanine was localized in intracellular vesicles. Vectorization by LDL favored lysosomal localization although little effect was found on the overall uptake as shown by extraction experiments. The role of lipoproteins in the cellular localization of photosensitizers is significantly more important for photosensitizers not freely diffusing through bilayers. The dynamics of the interactions of photosensitizers with membranes appears as an important determinant of their subcellular localization. PMID- 15345335 TI - Molecular mechanisms of ZD1839-induced G1-cell cycle arrest and apoptosis in human lung adenocarcinoma A549 cells. AB - Epithelial growth factor receptor (EGFR) has been proposed as a target for anticancer therapy. ZD1839 (Iressa) is a quinazoline derivative that selectively inhibits the EGFR tyrosine kinase activity and is under clinical use in cancer patients. However, the molecular mechanisms involved in ZD1839-mediated anticancer effects remain largely uncharacterized. In this study, exposure of human lung adenocarcinoma A549 cells to ZD1839 caused G1 arrest, and subsequently induced apoptosis. Moreover, ZD1839 increased the protein levels of p27(KIP1) and retinoblastoma-related Rb2/p130 while decreased the expression of cyclin dependent kinase-2 (CDK2), CDK4, CDK6 and cyclin-D1, cyclin-D3. In vitro kinase assay showed that ZD1839 decreased these CDKs expression in A549 cells, leading to significantly reduce their kinase activities. In addition, ZD1839-induced death of A549 cells with characteristics of apoptosis including apoptotic morphological changes, DNA fragmentation and enhancement of TUNEL-positive cell. These events were accompanied by a marked increase of Fas protein expression, and activation of caspase-2, -3, -8. Co-treatment of cells with Fas antagonist antibody significantly blocked ZD1839-induced apoptosis. Caspase-8 and caspase-3 inhibitors, but not a caspase-9 inhibitor, were also capable of restoring cell viability. Our results indicate that downregulation of the expression and function of CDK2, CDK4, CDK6, cyclin-D1 and cyclin-D3, as well as upregulation of p27(KIP1) and pRb2/p130, are strong candidates for the cell cycle regulator that arrests ZD1839-treated A549 cells at G1 phase. Furthermore, upregulation of Fas appears to play a major role in the initiation of ZD1839-induced apoptosis, activation of caspase-8/caspase-3 cascade is involved in the execution phase of this death program. PMID- 15345336 TI - Evaluation of glutathione deficiency in rat livers by microarray analysis. AB - Hepatic glutathione content was measured and gene expression data were obtained using an Affymetrix RG U34 array after treatment with tap water containing 20mM l buthionine (S, R)-sulfoximine (BSO) to male F344 rats for four consecutive days. Both Spearman's and Pearson's correlation coefficients were calculated between the glutathione content and the mRNA content level obtained from the microarray analysis individually. Sixty-nine gene probes, which were statistically significant (Spearman's correlation, P < 0.05) and showed a Pearson's correlation coefficients (Pearson's r) less than -0.8 between mRNA content and hepatic glutathione content, were identified as glutathione deficiency-correlated probes. By comparing the hepatic gene expression profiles between BSO- and butylated hydroxyanisole (BHA)-treated rats, 14 probes of genes that showed an increase in the corresponding gene mRNA levels only after the BSO treatment were thought to be good indicators of glutathione deficiency. A principal component analysis successfully illustrated the time-course of hepatic gene expression after the treatment with acetaminophen, phenobarbital and clofibrate, and the expression profiles were thought to reflect the changes in hepatic glutathione levels. The identified gene probes in the present study would be useful as markers for assessing hepatocellular glutathione deficiency, or oxidative stress level, based on microarray data. PMID- 15345337 TI - Dioxin revisited: developments since the 1997 IARC classification of dioxin as a human carcinogen. AB - In 1997 the International Agency for Research on Cancer (IARC) classified 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD; the most potent dioxin congener) as a group 1 carcinogen based on limited evidence in humans, sufficient evidence in experimental animals, and extensive mechanistic information indicating that TCDD acts through a mechanism involving the aryl hydrocarbon receptor (AhR), which is present in both humans and animals. The judgment of limited evidence in humans was based primarily on an elevation of all cancers combined in four industrial cohorts. The group 1 classification has been somewhat controversial and has been challenged in the literature in recent years. In this article we review the epidemiologic and mechanistic evidence that has emerged since 1997. New epidemiologic evidence consists primarily of positive exposure-response analyses in several of the industrial cohorts, as well as evidence of excesses of several specific cancers in the Seveso accident cohort. There are also new data regarding how the AhR functions in mediating the carcinogenic response to TCDD. The new evidence generally supports the 1997 IARC classification. PMID- 15345338 TI - The science and practice of carcinogen identification and evaluation. AB - Several national and international health agencies have established programs with the aim of identifying agents and exposures that cause cancer in humans. Carcinogen identification is an activity grounded in the scientific evaluation of the results of human epidemiologic studies, long-term bioassays in experimental animals, and other data relevant to an evaluation of carcinogenicity and its mechanisms. In this commentary, after a brief discussion of the science basis common to the evaluation of carcinogens across different programs, we discuss in more detail the principles and procedures currently used by the IARC Monographs program. PMID- 15345339 TI - Pesticide product use and risk of non-Hodgkin lymphoma in women. AB - A population-based, incidence case-control study was conducted among women in upstate New York to determine whether pesticide exposure is associated with an increase in risk of non-Hodgkin lymphoma (NHL) among women. The study involved 376 cases of NHL identified through the State Cancer Registry and 463 controls selected from the Medicare beneficiary files and state driver's license records. Information about history of farm work, history of other jobs associated with pesticide exposure, use of common household pesticide products, and potential confounding variables was obtained by telephone interview. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using an unconditional logistic regression model. The risk of NHL was doubled (OR = 2.12; 95% CI, 1.21-3.71) among women who worked for at least 10 years at a farm where pesticides were reportedly used. When both farming and other types of jobs associated with pesticide exposure were combined, there was a progressive increase in risk of NHL with increasing duration of such work (p = 0.005). Overall cumulative frequency of use of household pesticide products was positively associated with risk of NHL (p = 0.004), which was most pronounced when they were applied by subjects themselves. When exposure was analyzed by type of products used, a significant association was observed for mothballs. The associations with both occupational and household pesticides were particularly elevated if exposure started in 1950 1969 and for high-grade NHL. Although the results of this case-control study suggest that exposure to pesticide products may be associated with an increased risk of NHL among women, methodologic limitations related to selection and recall bias suggest caution in inferring causation. PMID- 15345340 TI - A Bayesian hierarchical approach for relating PM(2.5) exposure to cardiovascular mortality in North Carolina. AB - Considerable attention has been given to the relationship between levels of fine particulate matter (particulate matter < or = 2.5 microm in aerodynamic diameter; PM(2.5) in the atmosphere and health effects in human populations. Since the U.S. Environmental Protection Agency began widespread monitoring of PM(2.5) levels in 1999, the epidemiologic community has performed numerous observational studies modeling mortality and morbidity responses to PM(2.5) levels using Poisson generalized additive models (GAMs). Although these models are useful for relating ambient PM(2.5) levels to mortality, they cannot directly measure the strength of the effect of exposure to PM(2.5) on mortality. In order to assess this effect, we propose a three-stage Bayesian hierarchical model as an alternative to the classical Poisson GAM. Fitting our model to data collected in seven North Carolina counties from 1999 through 2001, we found that an increase in PM(2.5) exposure is linked to increased risk of cardiovascular mortality in the same day and next 2 days. Specifically, a 10- microg/m3 increase in average PM(2.5) exposure is associated with a 2.5% increase in the relative risk of current-day cardiovascular mortality, a 4.0% increase in the relative risk of cardiovascular mortality the next day, and an 11.4% increase in the relative risk of cardiovascular mortality 2 days later. Because of the small sample size of our study, only the third effect was found to have > 95% posterior probability of being > 0. In addition, we compared the results obtained from our model to those obtained by applying frequentist (or classical, repeated sampling-based) and Bayesian versions of the classical Poisson GAM to our study population. PMID- 15345341 TI - Serum dioxin concentrations and age at menarche. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a widespread environmental contaminant, is associated with delays in pubertal development in animal studies. On 10 July 1976, as a result of a chemical explosion, residents of Seveso, Italy, experienced the highest levels of TCDD exposure experienced by a human population. Twenty years later, we initiated the Seveso Women's Health Study (SWHS), a retrospective cohort study of female residents of the most contaminated areas, to determine whether the women were at higher risk for reproductive disease. We examined the association of TCDD serum levels, based on measurements in serum collected soon after the explosion, with reported age at menarche among the 282 SWHS women who were premenarcheal at the time of the explosion. We found no change in risk of onset of menarche with a 10-fold increase in TCDD (e.g., 10 100 ppt; hazard ratio = 0.95; 95% confidence interval, 0.83-1.09; p-value for trend = 0.46). When TCDD levels were categorized, there was also no evidence of a dose-response trend (p = 0.65). In summary, we found that individual serum TCDD measurements are not significantly related to age at menarche among women in the SWHS cohort. The women in this study experienced substantial TCDD exposure during the postnatal but prepubertal developmental period. Given that animal evidence suggests in utero exposure has the most significant effect on onset of puberty, continued follow-up of the offspring of the SWHS cohort is important. PMID- 15345342 TI - Exhaled breath condensate as a suitable matrix to assess lung dose and effects in workers exposed to cobalt and tungsten. AB - The aim of the present study was to investigate whether exhaled breath condensate (EBC), a fluid formed by cooling exhaled air, can be used as a suitable matrix to assess target tissue dose and effects of inhaled cobalt and tungsten, using EBC malondialdehyde (MDA) as a biomarker of pulmonary oxidative stress. Thirty-three workers exposed to Co and W in workshops producing either diamond tools or hard metal mechanical parts participated in this study. Two EBC and urinary samples were collected: one before and one at the end of the work shift. Controls were selected among nonexposed workers. Co, W, and MDA in EBC were analyzed with analytical methods based on mass spectrometric reference techniques. In the EBC from controls, Co was detectable at ultratrace levels, whereas W was undetectable. In exposed workers, EBC Co ranged from a few to several hundred nanomoles per liter. Corresponding W levels ranged from undetectable to several tens of nanomoles per liter. A parallel trend was observed for much higher urinary levels. Both Co and W in biological media were higher at the end of the work shift in comparison with preexposure values. In EBC, MDA levels were increased depending on Co concentration and were enhanced by coexposure to W. Such a correlation between EBC MDA and both Co and W levels was not observed with urinary concentration of either element. These results suggest the potential usefulness of EBC to complete and integrate biomonitoring and health surveillance procedures among workers exposed to mixtures of transition elements and hard metals. Key words: cobalt, exhaled breath condensate, hard metals, lung, malondialdehyde, oxidative stress, tungsten. PMID- 15345343 TI - Particulate matter exposure impairs systemic microvascular endothelium-dependent dilation. AB - Acute exposure to airborne pollutants, such as solid particulate matter (PM), increases the risk of cardiovascular dysfunction, but the mechanisms by which PM evokes systemic effects remain to be identified. The purpose of this study was to determine if pulmonary exposure to a PM surrogate, such as residual oil fly ash (ROFA), affects endothelium-dependent dilation in the systemic microcirculation. Rats were intratracheally instilled with ROFA at 0.1, 0.25, 1 or 2 mg/rat 24 hr before experimental measurements. Rats intratracheally instilled with saline or titanium dioxide (0.25 mg/rat) served as vehicle or particle control groups, respectively. In vivo microscopy of the spinotrapezius muscle was used to study systemic arteriolar dilator responses to the Ca2+ ionophore A23187, administered by ejection via pressurized micropipette into the arteriolar lumen. We used analysis of bronchoalveolar lavage (BAL) samples to monitor identified pulmonary inflammation and damage. To determine if ROFA exposure affected arteriolar nitric oxide sensitivity, sodium nitroprusside was iontophoretically applied to arterioles of rats exposed to ROFA. In saline-treated rats, A23187 dilated arterioles up to 72 +/- 7% of maximum. In ROFA- and TiO2-exposed rats, A23187 induced dilation was significantly attenuated. BAL fluid analysis revealed measurable pulmonary inflammation and damage after exposure to 1 and 2 mg ROFA (but not TiO2 or < 1 mg ROFA), as evidenced by significantly higher polymorphonuclear leukocyte cell counts, enhanced BAL albumin levels, and increased lactate dehydrogenase activity in BAL fluid. The sensitivity of arteriolar smooth muscle to NO was similar in saline-treated and ROFA-exposed rats, suggesting that pulmonary exposure to ROFA affected endothelial rather than smooth muscle function. A significant increase in venular leukocyte adhesion and rolling was observed in ROFA-exposed rats, suggesting local inflammation at the systemic microvascular level. These results indicate that pulmonary PM exposure impairs systemic endothelium-dependent arteriolar dilation. Moreover, because rats exposed to < 1 mg ROFA or TiO2 did not exhibit BAL signs of pulmonary damage or inflammation, it appears that PM exposure can impair systemic microvascular function independently of detectable pulmonary inflammation. PMID- 15345344 TI - Effects of low sulfur fuel and a catalyzed particle trap on the composition and toxicity of diesel emissions. AB - In this study we compared a "baseline" condition of uncontrolled diesel engine exhaust (DEE) emissions generated with current (circa 2003) certification fuel to an emissions-reduction (ER) case with low sulfur fuel and a catalyzed particle trap. Lung toxicity assessments (resistance to respiratory viral infection, lung inflammation, and oxidative stress) were performed on mice (C57Bl/6) exposed by inhalation (6 hr/day for 7 days). The engine was operated identically (same engine load) in both cases, and the inhalation exposures were conducted at the same exhaust dilution rate. For baseline DEE, this dilution resulted in a particle mass (PM) concentration of approximately 200 microg/m3 PM, whereas the ER reduced the PM and almost every other measured constituent [except nitrogen oxides (NOx)] to near background levels in the exposure atmospheres. These measurements included PM, PM size distribution, PM composition (carbon, ions, elements), NOx, carbon monoxide, speciated/total volatile hydrocarbons, and several classes of semivolatile organic compounds. After exposure concluded, one group of mice was immediately sacrificed and assessed for inflammation and oxidative stress in lung homogenate. Another group of mice were intratracheally instilled with respiratory syncytial virus (RSV), and RSV lung clearance and inflammation was assessed 4 days later. Baseline DEE produced statistically significant biological effects for all measured parameters. The use of low sulfur fuel and a catalyzed trap either completely or nearly eliminated the effects. PMID- 15345345 TI - Developmental dental aberrations after the dioxin accident in Seveso. AB - Children's developing teeth may be sensitive to environmental dioxins, and in animal studies developing teeth are one of the most sensitive targets of toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Twenty-five years after the dioxin accident in Seveso, Italy, 48 subjects from the contaminated areas (zones A and B) and in patches lightly contaminated (zone R) were recruited for the examination of dental and oral aberrations. Subjects were randomly invited from those exposed in their childhood and for whom frozen serum samples were available. The subjects were frequency matched with 65 subjects from the surrounding non-ABR zone for age, sex, and education. Concentrations of TCDD in previously analyzed plasma samples (zone ABR subjects only) ranged from 23 to 26,000 ng/kg in serum lipid. Ninety-three percent (25 of 27) of the subjects who had developmental enamel defects had been < 5 years of age at the time of the accident. The prevalence of defects in this age group was 42% (15 of 36) in zone ABR subjects and 26% (10 of 39) in zone non-ABR subjects, correlating with serum TCDD levels (p = 0.016). Hypodontia was seen in 12.5% (6 of 48) and 4.6% (3 of 65) of the zone ABR and non-ABR subjects, respectively, also correlating with serum TCDD level (p = 0.05). In conclusion, developmental dental aberrations were associated with childhood exposure to TCDD. In contrast, dental caries and periodontal disease, both infectious in nature, and oral pigmentation and salivary flow rate were not related to the exposure. The results support our hypothesis that dioxins can interfere with human organogenesis. PMID- 15345346 TI - Neurologic abnormalities in workers of a 1-bromopropane factory. AB - We reported recently that 1-bromopropane (1-BP; n-propylbromide, CAS Registry no. 106-94-5), an alternative to ozone-depleting solvents, is neurotoxic and exhibits reproductive toxicity in rats. The four most recent case reports suggested possible neurotoxicity of 1-BP in workers. The aim of the present study was to establish the neurologic effects of 1-BP in workers and examine the relationship with exposure levels. We surveyed 27 female workers in a 1-BP production factory and compared 23 of them with 23 age-matched workers in a beer factory as controls. The workers were interviewed and examined by neurologic, electrophysiologic, hematologic, biochemical, neurobehavioral, and postural sway tests. 1-BP exposure levels were estimated with passive samplers. Tests with a tuning fork showed diminished vibration sensation of the foot in 15 workers exposed to 1-BP but in none of the controls. 1-BP factory workers showed significantly longer distal latency in the tibial nerve than did the controls but no significant changes in motor nerve conduction velocity. Workers also displayed lower values in sensory nerve conduction velocity in the sural nerve, backward recalled digits, Benton visual memory test scores, pursuit aiming test scores, and five items of the Profile of Mood States (POMS) test (tension, depression, anxiety, fatigue, and confusion) compared with controls matched for age and education. Workers hired after May 1999, who were exposed to 1-BP only (workers hired before 1999 could have also been exposed to 2-BP), showed similar changes in vibration sense, distal latency, Benton test scores, and depression and fatigue in the POMS test. Time-weighted average exposure levels in the workers were 0.34-49.19 ppm. Exposure to 1-BP could adversely affect peripheral nerves or/and the central nervous system. PMID- 15345347 TI - Subcutaneous injection of mercury: "warding off evil". AB - Deliberate injection of mercury, especially subcutaneous injection, is rare but is seen in psychiatric patients, individuals who attempt suicide, those who are accidentally injected, and boxers who wish to build muscle bulk. Metallic mercury plays a major role in ethnic folk medicine. Neurologic and renal complications can result from high systemic levels of mercury, and subcutaneous injection usually results in sterile abscesses. Urgent surgical evacuation and close monitoring for neurologic and renal functions as well as chelation (if toxicity is indicated) are key aspects of treatment. Education of the adverse effects and dangers of mercury is important, especially in pregnant women and children. As increased immigration changes demographic patterns, proper disposal of mercury and preventing its sale and use should become urgent societal priorities. Psychiatric consultation should be obtained whenever appropriate. PMID- 15345348 TI - Water arsenic exposure and children's intellectual function in Araihazar, Bangladesh. AB - Exposure to arsenic has long been known to have neurologic consequences in adults, but to date there are no well-controlled studies in children. We report results of a cross-sectional investigation of intellectual function in 201 children 10 years of age whose parents participate in our ongoing prospective cohort study examining health effects of As exposure in 12,000 residents of Araihazar, Bangladesh. Water As and manganese concentrations of tube wells at each child's home were obtained by surveying all wells in the study region. Children and mothers came to our field clinic, where children received a medical examination in which weight, height, and head circumference were measured. Children's intellectual function on tests drawn from the Wechsler Intelligence Scale for Children, version III, was assessed by summing weighted items across domains to create Verbal, Performance, and Full-Scale raw scores. Children provided urine specimens for measuring urinary As and creatinine and were asked to provide blood samples for measuring blood lead and hemoglobin concentrations. Exposure to As from drinking water was associated with reduced intellectual function after adjustment for sociodemographic covariates and water Mn. Water As was associated with reduced intellectual function, in a dose-response manner, such that children with water As levels > 50 microg/L achieved significantly lower Performance and Full-Scale scores than did children with water As levels < 5.5 microg/L. The association was generally stronger for well-water As than for urinary As. PMID- 15345349 TI - Postweaning exposure to aflatoxin results in impaired child growth: a longitudinal study in Benin, West Africa. AB - Aflatoxins are dietary contaminants that are hepatocarcinogenic and immunotoxic and cause growth retardation in animals, but there is little evidence concerning the latter two parameters in exposed human populations. Aflatoxin exposure of West African children is known to be high, so we conducted a longitudinal study over an 8-month period in Benin to assess the effects of exposure on growth. Two hundred children 16-37 months of age were recruited from four villages, two with high and two with low aflatoxin exposure (50 children per village). Serum aflatoxin-albumin (AF-alb) adducts, anthropometric parameters, information on food consumption, and various demographic data were measured at recruitment (February) and at two subsequent time points (June and October). Plasma levels of vitamin A and zinc were also measured. AF-alb adducts increased markedly between February and October in three of the four villages, with the largest increases in the villages with higher exposures. Children who were fully weaned at recruitment had higher AF-alb than did those still partially breast-fed (p < 0.0001); the major weaning food was a maize-based porridge. There was no association between AF-alb and micronutrient levels, suggesting that aflatoxin exposure was not accompanied by a general nutritional deficiency. There was, however, a strong negative correlation (p < 0.0001) between AF-alb and height increase over the 8 month follow-up after adjustment for age, sex, height at recruitment, socioeconomic status, village, and weaning status; the highest quartile of AF-alb was associated with a mean 1.7 cm reduction in growth over 8 months compared with the lowest quartile. This study emphasizes the association between aflatoxin and stunting, although the underlying mechanisms remain unclear. Aflatoxin exposure during the weaning period may be critical in terms of adverse health effects in West African children, and intervention measures to reduce exposure merit investigation. PMID- 15345350 TI - Follow-up study of adolescents exposed to di(2-ethylhexyl) phthalate (DEHP) as neonates on extracorporeal membrane oxygenation (ECMO) support. AB - Di(2-ethylhexyl) phthalate (DEHP) is used to make polyvinyl chloride (PVC) plastic tubing soft and flexible. Animal data show that adverse effects of DEHP exposure may include reduced fertility, reduced sperm production in males, and ovarian dysfunction in females. Known treatments that involve high DEHP exposures are blood exchange transfusions, extracorporeal membrane oxygenation (ECMO), and cardiovascular surgery. Although potential exposure to DEHP in ECMO patients is significant, the exposure has not been associated with short-term toxicity. To evaluate long-term toxicity, we undertook a study of neonatal ECMO survivors to assess their onset of puberty and sexual maturity. We evaluated 13 male and 6 female subjects at 14-16 years of age who had undergone ECMO as neonates. All subjects had a complete physical examination including measurements for height, weight, head circumference, and pubertal assessment by Tanner staging. The testicular volume and the phallic length were measured in male participants. Laboratory tests included thyroid, liver, and renal function as well as measurements of luteinizing hormone, follicle-stimulating hormone, testosterone for males, and estradiol for females. Except for one patient with Marfan syndrome, the rest had normal growth percentile for age and sex. All had normal values for thyroid, liver, and renal functions. Sexual hormones were appropriate for the stage of pubertal maturity. Our results indicate that adolescents exposed to significant quantities of DEHP as neonates showed no significant adverse effects on their physical growth and pubertal maturity. Thyroid, liver, renal, and male and female gonadal functions tested were within normal range for age and sex distribution. Key Words: DEHP, ECMO, toxicity. PMID- 15345351 TI - Breast milk: an optimal food. PMID- 15345352 TI - Activities and organophosphate exposures: need for the numbers. PMID- 15345353 TI - Prenatal lead exposure and schizophrenia: a plausible neurobiologic connection. PMID- 15345355 TI - Electromagnetic fields and free radicals. PMID- 15345357 TI - Complexity of factors involved in human population growth. PMID- 15345358 TI - Past and future considerations for heavy-duty diesel engine emissions. PMID- 15345360 TI - Monitoring for asbestos: U.S. EPA methods. PMID- 15345361 TI - Evaluating the toxicity of chemical mixtures. PMID- 15345362 TI - Electronics, lead, and landfills. PMID- 15345363 TI - Beyond the bench: Fish tales to ensure health. PMID- 15345364 TI - Nanotechnology: looking as we leap. PMID- 15345365 TI - Botanical supplements: weeding out the health risks. PMID- 15345366 TI - New spin on an old fiber. PMID- 15345368 TI - Gene interaction network suggests dioxin induces a significant linkage between aryl hydrocarbon receptor and retinoic acid receptor beta. AB - Gene expression arrays (gene chips) have enabled researchers to roughly quantify the level of mRNA expression for a large number of genes in a single sample. Several methods have been developed for the analysis of gene array data including clustering, outlier detection, and correlation studies. Most of these analyses are aimed at a qualitative identification of what is different between two samples and/or the relationship between two genes. We propose a quantitative, statistically sound methodology for the analysis of gene regulatory networks using gene expression data sets. The method is based on Bayesian networks for direct quantification of gene expression networks. Using the gene expression changes in HPL1A lung airway epithelial cells after exposure to 2,3,7,8 tetrachlorodibenzo-(Italic)p(/Italic)-dioxin at levels of 0.1, 1.0, and 10.0 nM for 24 hr, a gene expression network was hypothesized and analyzed. The method clearly demonstrates support for the assumed network and the hypothesis linking the usual dioxin expression changes to the retinoic acid receptor system. Simulation studies demonstrated the method works well, even for small samples. PMID- 15345369 TI - Valproic acid teratogenicity: a toxicogenomics approach. AB - Embryonic development is a highly coordinated set of processes that depend on hierarchies of signaling and gene regulatory networks, and the disruption of such networks may underlie many cases of chemically induced birth defects. The antiepileptic drug valproic acid (VPA) is a potent inducer of neural tube defects (NTDs) in human and mouse embryos. As with many other developmental toxicants however, the mechanism of VPA teratogenicity is unknown. Using microarray analysis, we compared the global gene expression responses to VPA in mouse embryos during the critical stages of teratogen action in vivo with those in cultured P19 embryocarcinoma cells in vitro. Among the identified VPA-responsive genes, some have been associated previously with NTDs or VPA effects [vinculin, metallothioneins 1 and 2 (Mt1, Mt2), keratin 1-18 (Krt1-18)], whereas others provide novel putative VPA targets, some of which are associated with processes relevant to neural tube formation and closure [transgelin 2 (Tagln2), thyroid hormone receptor interacting protein 6, galectin-1 (Lgals1), inhibitor of DNA binding 1 (Idb1), fatty acid synthase (Fasn), annexins A5 and A11 (Anxa5, Anxa11)], or with VPA effects or known molecular actions of VPA (Lgals1, Mt1, Mt2, Id1, Fasn, Anxa5, Anxa11, Krt1-18). A subset of genes with a transcriptional response to VPA that is similar in embryos and the cell model can be evaluated as potential biomarkers for VPA-induced teratogenicity that could be exploited directly in P19 cell-based in vitro assays. As several of the identified genes may be activated or repressed through a pathway of histone deacetylase (HDAC) inhibition and specificity protein 1 activation, our data support a role of HDAC as an important molecular target of VPA action in vivo. PMID- 15345370 TI - Discriminating different classes of toxicants by transcript profiling. AB - Male rats were treated with various model compounds or the appropriate vehicle controls. Most substances were either well-known hepatotoxicants or showed hepatotoxicity during preclinical testing. The aim of the present study was to determine if biological samples from rats treated with various compounds can be classified based on gene expression profiles. In addition to gene expression analysis using microarrays, a complete serum chemistry profile and liver and kidney histopathology were performed. We analyzed hepatic gene expression profiles using a supervised learning method (support vector machines; SVMs) to generate classification rules and combined this with recursive feature elimination to improve classification performance and to identify a compact subset of probe sets with potential use as biomarkers. Two different SVM algorithms were tested, and the models obtained were validated with a compound based external cross-validation approach. Our predictive models were able to discriminate between hepatotoxic and nonhepatotoxic compounds. Furthermore, they predicted the correct class of hepatotoxicant in most cases. We provide an example showing that a predictive model built on transcript profiles from one rat strain can successfully classify profiles from another rat strain. In addition, we demonstrate that the predictive models identify nonresponders and are able to discriminate between gene changes related to pharmacology and toxicity. This work confirms the hypothesis that compound classification based on gene expression data is feasible. PMID- 15345371 TI - Assessment of prediction confidence and domain extrapolation of two structure activity relationship models for predicting estrogen receptor binding activity. AB - Quantitative structure-activity relationship (QSAR) methods have been widely applied in drug discovery, lead optimization, toxicity prediction, and regulatory decisions. Despite major advances in algorithms and software, QSAR models have inherent limitations associated with a size and chemical-structure diversity of the training set, experimental error, and many characteristics of structure representation and correlation algorithms. Whereas excellent fit to the training data may be readily attainable, often models fail to predict accurately chemicals that are outside their domain of applicability. A QSAR's utility and, in the case of regulatory decisions, justification for usage increasingly depend on the ability to quantify a model's potential for predicting unknown chemicals with some known degree of certainty. It is never possible to predict an unknown chemical with absolute certainty. Here we report on two QSAR models based on different data sets for classification of chemicals according to their ability to bind to the estrogen receptor. The models were developed by using a novel QSAR method, Decision Forest, which combines the results of multiple heterogeneous but comparable Decision Tree models to produce a consensus prediction. We used an extensive cross-validation process to define an applicability domain for model predictions based on two quantitative measures: prediction confidence and domain extrapolation. Together, these measures quantify the accuracy of each prediction within and outside of the training domain. Despite being based on large and diverse training sets, both QSAR models had poor accuracy for chemicals within the domain of low confidence, whereas good accuracy was obtained for those within the domain of high confidence. For prediction in the high confidence domain, accuracy was inversely proportional to the degree of domain extrapolation. The model with a larger training set of 1,092, compared with 232 for the other, was more accurate in predicting chemicals at larger domain extrapolation, and could be particularly useful for rapidly prioritizing potential endocrine disruptors from large chemical universe. PMID- 15345372 TI - Biokinetics and subchronic toxic effects of oral arsenite, arsenate, monomethylarsonic acid, and dimethylarsinic acid in v-Ha-ras transgenic (Tg.AC) mice. AB - Previous research demonstrated that 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment increased the number of skin papillomas in v-Ha-ras transgenic (Tg.AC) mice that had received sodium arsenite [(As(III)] in drinking water, indicating that this model is useful for studying the toxic effects of arsenic in vivo. Because the liver is a known target of arsenic, we examined the pathophysiologic and molecular effects of inorganic and organic arsenical exposure on Tg.AC mouse liver in this study. Tg.AC mice were provided drinking water containing As(III), sodium arsenate [As(V)], monomethylarsonic acid [(MMA(V)], and 1,000 ppm dimethylarsinic acid [DMA(V)] at dosages of 150, 200, 1,500, or 1,000 ppm as arsenic, respectively, for 17 weeks. Control mice received unaltered water. Four weeks after initiation of arsenic treatment, TPA at a dose of 1.25 microg/200 microL acetone was applied twice a week for 2 weeks to the shaved dorsal skin of all mice, including the controls not receiving arsenic. In some cases arsenic exposure reduced body weight gain and caused mortality (including moribundity). Arsenical exposure resulted in a dose-dependent accumulation of arsenic in the liver that was unexpectedly independent of chemical species and produced hepatic global DNA hypomethylation. cDNA microarray and reverse transcriptase-polymerase chain reaction analysis revealed that all arsenicals altered the expression of numerous genes associated with toxicity and cancer. However, organic arsenicals [MMA(V) and DMA(V)] induced a pattern of gene expression dissimilar to that of inorganic arsenicals. In summary, subchronic exposure of Tg.AC mice to inorganic or organic arsenicals resulted in toxic manifestations, hepatic arsenic accumulation, global DNA hypomethylation, and numerous gene expression changes. These effects may play a role in arsenic-induced hepatotoxicity and carcinogenesis and may be of particular toxicologic relevance. PMID- 15345373 TI - Toxicogenomics in risk assessment: communicating the challenges. PMID- 15345374 TI - Regulatory acceptance of toxicogenomics data. PMID- 15345376 TI - Standards for microarray data: an open letter. PMID- 15345377 TI - Encyclopedia of DNA. PMID- 15345378 TI - Cooperation achieves results at UNC-CH. PMID- 15345379 TI - Toxicogenomics data: the road to acceptance. AB - Regulatory agencies believe microarray data could be extremely useful in testing prospective chemical products and investigating safety problems after chemicals have been marketed. However, it has not yet been clearly established how this information will be used by agencies in the approval of pharmaceuticals and other chemicals--or even whether companies will be required to submit microarray data. Although some private companies are already voluntarily submitting microarray data along with their drug and pesticide applications, others are hesitant to do so. Most of the stakeholders involved agree that standardization of microarray experiment procedures and of genomic signatures are key to the broad acceptance and use of these data. PMID- 15345382 TI - Comparison of diversities and compositions of bacterial populations inhabiting natural forest soils. AB - The diversity and composition of soil bacterial communities were compared among six Austrian natural forests, including oak-hornbeam, spruce-fir-beech, and Austrian pine forests, using terminal restriction fragment length polymorphism (T RFLP, or TRF) analysis and sequence analysis of 16S rRNA genes. The forests studied differ greatly in soil chemical characteristics, microbial biomass, and nutrient turnover rates. The aim of this study was to relate these differences to the composition of the bacterial communities inhabiting the individual forest soils. Both TRF profiling and clone sequence analysis revealed that the bacterial communities in soils under Austrian pine forests, representing azonal forest types, were distinct from those in soils under zonal oak-hornbeam and spruce-fir beech forests, which were more similar in community composition. Clones derived from an Austrian pine forest soil were mostly affiliated with high-G+C gram positive bacteria (49%), followed by members of the alpha-Proteobacteria (20%) and the Holophaga/Acidobacterium group (12%). Clones in libraries from oak hornbeam and spruce-fir-beech forest soils were mainly related to the Holophaga/Acidobacterium group (28 and 35%), followed by members of the Verrucomicrobia (24%) and the alpha-Proteobacteria (27%), respectively. The soil bacterial communities in forests with distinct vegetational and soil chemical properties appeared to be well differentiated based on 16S rRNA gene phylogeny. In particular, the outstanding position of the Austrian pine forests, which are determined by specific soil conditions, was reflected in the bacterial community composition. PMID- 15345383 TI - Characterization and heterologous expression of the oxalyl coenzyme A decarboxylase gene from Bifidobacterium lactis. AB - Oxalyl coenzyme A (CoA) decarboxylase (Oxc) is a key enzyme in the catabolism of the highly toxic compound oxalate, catalyzing the decarboxylation of oxalyl-CoA to formyl-CoA. The gene encoding a novel oxalyl-CoA decarboxylase from Bifidobacterium lactis DSM 10140 (oxc) was identified and characterized. This strain, isolated from yogurt, showed the highest oxalate-degrading activity in a preliminary screening with 12 strains belonging to Bifidobacterium, an anaerobic intestinal bacterial group largely used in probiotic products. The oxc gene was isolated by probing a B. lactis genomic library with a probe obtained by amplification of the oxalyl-CoA decarboxylase gene from Oxalobacter formigenes, an anaerobic bacterium of the human intestinal microflora. The oxc DNA sequence analysis revealed an open reading frame of 1,773 bp encoding a deduced 590-amino acid protein with a molecular mass of about 63 kDa. Analysis of amino acid sequence showed a significant homology (47%) with oxalyl-CoA decarboxylase of O. formigenes and a typical thiamine pyrophosphate-binding site that has been reported for several decarboxylase enzymes. Primer extension experiments with oxc performed by using RNA isolated from B. lactis identified the transcriptional start site 28 bp upstream of the ATG start codon, immediately adjacent to a presumed promoter region. The protein overexpressed in Escherichia coli cross reacted with an anti-O. formigenes oxalyl-CoA decarboxylase antibody. Enzymatic activity, when evaluated by capillary electrophoresis analysis, demonstrated that the consumption substrate oxalyl-CoA was regulated by a product inhibition of the enzyme. These findings suggest a potential role for Bifidobacterium in the intestinal degradation of oxalate. PMID- 15345384 TI - Display of bacterial lipase on the Escherichia coli cell surface by using FadL as an anchoring motif and use of the enzyme in enantioselective biocatalysis. AB - We have developed a novel cell surface display system by employing FadL as an anchoring motif, which is an outer membrane protein involved in long-chain fatty acid transport in Escherichia coli. A thermostable Bacillus sp. strain TG43 lipase (44.5 kDa) could be successfully displayed on the cell surface of E. coli in an active form by C-terminal deletion-fusion of lipase at the ninth external loop of FadL. The localization of the truncated FadL-lipase fusion protein on the cell surface was confirmed by confocal microscopy and Western blot analysis. Lipase activity was mainly detected with whole cells, but not with the culture supernatant, suggesting that cell lysis was not a problem. The activity of cell surface-displayed lipase was examined at different temperatures and pHs and was found to be the highest at 50 degrees C and pH 9 to 10. Cell surface-displayed lipase was quite stable, even at 60 and 70 degrees C, and retained over 90% of the full activity after incubation at 50 degrees C for a week. As a potential application, cell surface-displayed lipase was used as a whole-cell catalyst for kinetic resolution of racemic methyl mandelate. In 36 h of reaction, (S)-mandelic acid could be produced with the enantiomeric excess of 99% and the enantiomeric ratio of 292, which are remarkably higher than values obtained with crude lipase or cross-linked lipase crystal. These results suggest that FadL may be a useful anchoring motif for displaying enzymes on the cell surface of E. coli for whole cell biocatalysis. PMID- 15345385 TI - Influence of complex nutrient source on growth of and curvacin a production by sausage isolate Lactobacillus curvatus LTH 1174. AB - Lactobacillus curvatus LTH 1174, a fermented sausage isolate, produces the antilisterial bacteriocin curvacin A. Its biokinetics of cell growth and bacteriocin production as a function of various concentrations of a complex nutrient source were investigated in vitro during laboratory fermentations with modified MRS medium. A modification of the nutrient depletion model was used to fit the data describing growth and bacteriocin production. Both cell growth and bacteriocin activity were influenced by changes in the complex nutrient source concentration. Standard MRS medium clearly limited the growth of L. curvatus LTH 1174. Higher nutrient concentrations, up to a certain degree, led to improved growth, a higher attainable biomass concentration, and a higher bacteriocin activity in the supernatant. A lower concentration of complex nutrient source caused severe growth inhibition, leading to a lower biomass concentration but a much higher specific bacteriocin production. When examining the separate components of the complex nutrient source, a stimulating effect of bacteriological peptone on growth was found without an adverse effect on bacteriocin production, resulting in increased curvacin A activity. Furthermore, specific depletion of the amino acids tyrosine, serine, and asparagine/aspartic acid was observed for this strain. PMID- 15345386 TI - Calicivirus inactivation by nonionizing (253.7-nanometer-wavelength [UV]) and ionizing (gamma) radiation. AB - Noroviruses (previously Norwalk-like viruses) are the most common viral agents associated with food- and waterborne outbreaks of gastroenteritis. In the absence of culture methods for noroviruses, animal caliciviruses were used as model viruses to study inactivation by nonionizing (253.7-nm-wavelength [UV]) and ionizing (gamma) radiation. Here, we studied the respiratory feline calicivirus (FeCV) and the presumed enteric canine calicivirus (CaCV) and compared them with the well-studied bacteriophage MS2. When UV irradiation was used, a 3-log(10) reduction was observed at a fluence of 120 J/m(2) in the FeCV suspension and at a fluence of 200 J/m(2) for CaCV; for the more resistant phage MS2 there was a 3 log(10) reduction at a fluence of 650 J/m(2). Few or no differences were observed between levels of UV inactivation in high- and low-protein-content virus stocks. In contrast, ionizing radiation could readily inactivate MS2 in water, and there was a 3-log(10) reduction at a dose of 100 Gy, although this did not occur when the phage was diluted in high-protein-content stocks of CaCV or FeCV. The low protein-content stocks showed 3-log(10) reductions at a dose of 500 Gy for FeCV and at a dose of 300 for CaCV. The inactivation rates for both caliciviruses with ionizing and nonionizing radiation were comparable but different from the inactivation rates for MS2. Although most FeCV and CaCV characteristics, such as overall particle and genome size and structure, are similar, the capsid sequences differ significantly, making it difficult to predict human norovirus inactivation. Adequate management of UV and gamma radiation processes for virus inactivation should limit public health risks. PMID- 15345387 TI - Diversity and seasonal changes of uncultured Planctomycetales in river biofilms. AB - Cell counts of planctomycetes showed that there were high levels of these organisms in the summer and low levels in the winter in biofilms grown in situ in two polluted rivers, the Elbe River and the Spittelwasser River. In this study 16S rRNA-based methods were used to investigate if these changes were correlated with changes in the species composition. Planctomycete-specific clone libraries of the 16S rRNA genes found in both rivers showed that there were seven clusters, which were distantly related to the genera Pirellula, Planctomyces, and Gemmata. The majority of the sequences from the Spittelwasser River were affiliated with a cluster related to Pirellula, while the majority of the clones from the Elbe River fell into three clusters related to Planctomyces and one deeply branching cluster related to Pirellula. Some clusters also contained sequences derived from freshwater environments worldwide, and the similarities to our biofilm clones were as high as 99.8%, indicating the presence of globally distributed freshwater clusters of planctomycetes that have not been cultivated yet. Community fingerprints of planctomycete 16S rRNA genes were generated by temperature gradient gel electrophoresis from Elbe River biofilm samples collected monthly for 1 year. Sixteen bands were identified, and for the most part these bands represented organisms related to the genus Planctomyces. The fingerprints showed that there was strong seasonality of most bands and that there were clear differences in the summer and the winter. Thus, seasonal changes in the abundance of Planctomycetales in river biofilms were coupled to shifts in the community composition. PMID- 15345388 TI - The gnyRDBHAL cluster is involved in acyclic isoprenoid degradation in Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa PAO1 mutants affected in the ability to degrade acyclic isoprenoids were isolated with transposon mutagenesis. The gny cluster (for geranoyl), which encodes the enzymes involved in the lower pathway of acyclic isoprenoid degradation, was identified. The gny cluster is constituted by five probable structural genes, gnyDBHAL, and a possible regulatory gene, gnyR. Mutations in the gnyD, gnyB, gnyA, or gnyL gene caused inability to assimilate acyclic isoprenoids of the citronellol family of compounds. Transcriptional analysis showed that expression of the gnyB gene was induced by citronellol and repressed by glucose, whereas expression of the gnyR gene had the opposite behavior. Western blot analysis of citronellol-grown cultures showed induction of biotinylated proteins of 70 and 73 kDa, which probably correspond to 3 methylcrotonoyl-coenzyme A (CoA) carboxylase and geranoyl-CoA carboxylase (GCCase) alpha subunits, respectively. The 73-kDa biotinylated protein, identified as the alpha-GCCase subunit, is encoded by gnyA. Intermediary metabolites of the isoprenoid pathway, citronellic and geranic acids, were shown to accumulate in gnyB and gnyA mutants. Our data suggest that the protein products encoded in the gny cluster are the beta and alpha subunits of geranoyl CoA carboxylase (GnyB and GnyA), the citronelloyl-CoA dehydrogenase (GnyD), the gamma-carboxygeranoyl-CoA hydratase (GnyH), and the 3-hydroxy-gamma carboxygeranoyl-CoA lyase (GnyL). We conclude that the gnyRDBHAL cluster is involved in isoprenoid catabolism. PMID- 15345389 TI - Effect of length of time before incorporation on survival of pathogenic bacteria present in livestock wastes applied to agricultural soil. AB - In response to reports that the contamination of food can occur during the on farm primary phase of food production, we report data that describes a possible cost-effective intervention measure. The effect of time before soil incorporation of livestock wastes spread to land on the rate of decline of zoonotic agents present in the waste was investigated. Fresh livestock wastes were inoculated with laboratory-cultured Salmonella, Listeria, and Campylobacter spp. and Escherichia coli O157 before they were spread onto soil. Incorporation of the spread wastes was either immediate, delayed for 1 week, or did not occur at all. Bacterial decline was monitored over time and found to be significantly more rapid for all waste types when they were left on the soil surface. There were no significant differences in initial bacterial decline rates when wastes were spread in summer or winter. Our results indicate that not incorporating contaminated livestock wastes into soil is a potential intervention measure that may help to limit the spread of zoonotic agents further up the food chain. The implications of these findings are discussed in relation to current advice for livestock waste disposal. PMID- 15345390 TI - Comparison of ATPase-encoding type III secretion system hrcN genes in biocontrol fluorescent Pseudomonads and in phytopathogenic proteobacteria. AB - Type III protein secretion systems play a key role in the virulence of many pathogenic proteobacteria, but they also occur in nonpathogenic, plant-associated bacteria. Certain type III protein secretion genes (e.g., hrcC) have been found in Pseudomonas sp. strain SBW25 (and other biocontrol pseudomonads), but other type III protein secretion genes, such as the ATPase-encoding gene hrcN, have not been found. Using both colony hybridization and a PCR approach, we show here that hrcN is nevertheless present in many biocontrol fluorescent pseudomonads. The phylogeny of biocontrol Pseudomonas strains based on partial hrcN sequences was largely congruent with the phylogenies derived from analyses of rrs (encoding 16S rRNA) and, to a lesser extent, biocontrol genes, such as phlD (for 2,4 diacetylphloroglucinol production) and hcnBC (for HCN production). Most biocontrol pseudomonads clustered separately from phytopathogenic proteobacteria, including pathogenic pseudomonads, in the hrcN tree. The exception was strain KD, which clustered with phytopathogenic pseudomonads, such as Pseudomonas syringae, suggesting that hrcN was acquired from the latter species. Indeed, strain KD (unlike strain SBW25) displayed the same organization of the hrpJ operon, which contains hrcN, as P. syringae. These results indicate that the occurrence of hrcN in most biocontrol pseudomonads is not the result of recent horizontal gene transfer from phytopathogenic bacteria, although such transfer might have occurred for a minority of biocontrol strains. PMID- 15345391 TI - Clonality and diversity of the fish pathogen Lactococcus garvieae in Mediterranean countries. AB - Infection with Lactococcus garvieae is considered the most important risk factor for the European trout industry, and the losses are approximately 50% of the total production. To improve our understanding of the genetic links among strains originating from different countries, we examined the population structure of L. garvieae by comparing 81 strains isolated from different sources and ecosystems (41 farms in six countries) in which the bacterium is commonly found. Genetic similarities (as assessed with molecular tools, including restriction fragment length polymorphism ribotyping with two endonucleases) were compared with serological data. The combined results reveal that in endemic sites the bacterial population displays a clonal structure, whereas bacterial diversity characterizes sites where the infection is sporadic. PMID- 15345392 TI - Surveillance of bulk raw and commercially pasteurized cows' milk from approved Irish liquid-milk pasteurization plants to determine the incidence of Mycobacterium paratuberculosis. AB - Over the 13-month period from October 2000 to November 2001 (inclusive), the Food Safety Authority of Ireland (FSAI) carried out surveillance of Irish bulk raw (n = 389) and commercially pasteurized (n = 357) liquid-milk supplies to determine the incidence of Mycobacterium paratuberculosis. The pasteurization time temperature conditions were recorded for all pasteurized samples. Overall, 56% of whole-milk pasteurized samples had been heat treated at or above a time temperature combination of 75 degrees C for 25 s. All analyses were undertaken at the Department of Food Science (Food Microbiology) laboratory at Queen's University Belfast. Each milk sample was subjected to two tests for M. paratuberculosis: immunomagnetic separation-PCR (IMS-PCR; to detect the presence of M. paratuberculosis cells, live or dead) and chemical decontamination and culture (to confirm the presence of viable M. paratuberculosis). Overall, M. paratuberculosis DNA was detected by IMS-PCR in 50 (12.9%; 95% confidence interval, 9.9 to 16.5%) raw-milk samples and 35 (9.8%; 95% confidence interval, 7.1 to 13.3%) pasteurized-milk samples. Confirmed M. paratuberculosis was cultured from one raw-milk sample and no pasteurized-milk samples. It is concluded that M. paratuberculosis DNA is occasionally present at low levels in both raw and commercially pasteurized cows' milk. However, since no viable M. paratuberculosis was isolated from commercially pasteurized cows' milk on retail sale in the Republic of Ireland, current pasteurization procedures are considered to be effective. PMID- 15345393 TI - Modulation of Aspergillus awamori thaumatin secretion by modification of bipA gene expression. AB - Two different strains, Aspergillus awamori TGDTh-4 and A. awamori TGP-3 overexpressing a synthetic gene encoding the plant sweet protein thaumatin, showed an unfolded protein response. To facilitate protein secretion, the chaperone BiPA gene was expressed in A. awamori under control of the strong constitutive promoter of the gpdA gene. A good correlation was observed between the level of the bipA transcript in different strains and the amount of thaumatin secreted. Thaumatin secretion was increased 2- to 2.5-fold in transformants overexpressing the bipA gene compared with the parental strain. Secretion of the homologous proteins alpha-amylase and glucoamylase was not affected by the bipA gene overexpression. The requirement for BiPA for secretion of thaumatin was confirmed by attenuation of the endogenous bipA gene expression with an antisense RNA cassette. The decrease in bipA expression reduced the amount of secreted thaumatin up to 80% without affecting the secretion of the homologous alpha amylase and glucoamylase proteins. The BiPA protein is, therefore, very important for secretion of some heterologous proteins, such as thaumatin in A. awamori. PMID- 15345394 TI - Pulsed-field gel electrophoresis analysis of Vibrio vulnificus strains isolated from Taiwan and the United States. AB - Vibrio vulnificus is a marine bacterium that causes human wound infections and septicemia with a high mortality rate. V. vulnificus strains from different clinical and environmental sources or geographic regions have been successfully characterized by ribotyping and several other methods. Pulsed-field gel electrophoresis (PFGE) is a highly discriminative method, but previous studies suggested that it was not suitable for examining the correlation of V. vulnificus strains from different origins. We employed PFGE to determine its efficacy for characterizing V. vulnificus strains from different geographic regions, characterizing a total of 153 strains from clinical and environmental origins from the United States and Taiwan after SfiI or NotI digestion. V. vulnificus strains showed a high intraspecific diversity by PFGE after SfiI or NotI digestion, and about 12% of the strains could not be typed by the use of either of these enzymes. For PFGE with SfiI digestion, most of the clinical and environmental strains from the United States were grouped into cluster A, while the strains from Taiwan were grouped into other clusters. Clinical strains from the United States showed a higher level of genetic homogeneity than clinical strains from Taiwan, and environmental strains from both regions showed a similarly high level of heterogeneity. PFGE with NotI digestion was useful for studying the correlation of clinical strains from the United States and Taiwan, but it was not suitable for analyzing environmental strains. The results showed that PFGE with SfiI digestion may be used to characterize V. vulnificus strains from distant geographic regions, with NotI being a recommended alternative enzyme. PMID- 15345395 TI - Assessment of production conditions for efficient use of Escherichia coli in high yield heterologous recombinant selenoprotein synthesis. AB - The production of heterologous selenoproteins in Escherichia coli necessitates the design of a secondary structure in the mRNA forming a selenocysteine insertion sequence (SECIS) element compatible with SelB, the elongation factor for selenocysteine insertion at a predefined UGA codon. SelB competes with release factor 2 (RF2) catalyzing translational termination at UGA. Stoichiometry between mRNA, the SelB elongation factor, and RF2 is thereby important, whereas other expression conditions affecting the yield of recombinant selenoproteins have been poorly assessed. Here we expressed the rat selenoprotein thioredoxin reductase, with titrated levels of the selenoprotein mRNA under diverse growth conditions, with or without cotransformation of the accessory bacterial selA, selB, and selC genes. Titration of the selenoprotein mRNA with a pBAD promoter was performed in both TOP10 and BW27783 cells, which unexpectedly could not improve yield or specific activity compared to that achieved in our prior studies. Guided by principal component analysis, we instead discovered that the most efficient bacterial selenoprotein production conditions were obtained with the high-transcription T7lac-driven pET vector system in presence of the selA, selB, and selC genes, with induction of production at late exponential phase. About 40 mg of rat thioredoxin reductase with 50% selenocysteine content could thereby be produced per liter bacterial culture. These findings clearly illustrate the ability of E. coli to upregulate the selenocysteine incorporation machinery on demand and that this is furthermore strongly augmented in late exponential phase. This study also demonstrates that E. coli can indeed be utilized as cell factories for highly efficient production of heterologous selenoproteins such as rat thioredoxin reductase. PMID- 15345396 TI - Genotyping and toxigenic potential of Bacillus subtilis and Bacillus pumilus strains occurring in industrial and artisanal cured sausages. AB - Artisanal and industrial sausages were analyzed for their aerobic, heat-resistant microflora to assess whether new emerging pathogens could be present among Bacillus strains naturally contaminating cured meat products. Sixty-four isolates were characterized by randomly amplified polymorphic DNA (RAPD)-PCR and fluorescent amplified fragment length polymorphism (fAFLP). The biotypes, identified by partial 16S rRNA gene sequence analysis, belonged to Bacillus subtilis, Bacillus pumilus, and Bacillus amyloliquefaciens species. Both RAPD-PCR and fAFLP analyses demonstrated that a high genetic heterogeneity is present in the B. subtilis group even in strains harvested from the same source, making it possible to isolate 56 different biotypes. Moreover, fAFLP analysis made it possible to distinguish B. subtilis from B. pumilus strains. The strains were characterized for their toxigenic potential by molecular, physiological, and immunological techniques. Specific PCR analyses revealed the absence of DNA sequences related to HBL, BcET, NHE, and entFM Bacillus cereus enterotoxins and the enzymes sphingomyelinase Sph and phospholipase PI-PLC in all strains; also, the immunological analyses showed that Bacillus strains did not react with NHE- and HBL-specific antibodies. However, some isolates were found to be positive for hemolytic and lecithinase activity. The absence of toxigenic potential in Bacillus strains from the sausages analyzed indicates that these products can be considered safe under the processing conditions they were produced; however, great care should be taken when the ripening time is shortened, particularly in the case of traditional sausages, which could contain high amounts of Bacillus strains and possibly some B. cereus cells. PMID- 15345397 TI - Copper ions stimulate polyphosphate degradation and phosphate efflux in Acidithiobacillus ferrooxidans. AB - For some bacteria and algae, it has been proposed that inorganic polyphosphates and transport of metal-phosphate complexes could participate in heavy metal tolerance. To test for this possibility in Acidithiobacillus ferrooxidans, a microorganism with a high level of resistance to heavy metals, the polyphosphate levels were determined when the bacterium was grown in or shifted to the presence of a high copper concentration (100 mM). Under these conditions, cells showed a rapid decrease in polyphosphate levels with a concomitant increase in exopolyphosphatase activity and a stimulation of phosphate efflux. Copper in the range of 1 to 2 microM greatly stimulated exopolyphosphatase activity in cell extracts from A. ferrooxidans. The same was seen to a lesser extent with cadmium and zinc. Bioinformatic analysis of the available A. ferrooxidans ATCC 23270 genomic sequence did not show a putative pit gene for phosphate efflux but rather an open reading frame similar in primary and secondary structure to that of the Saccharomyces cerevisiae phosphate transporter that is functional at acidic pH (Pho84). Our results support a model for metal detoxification in which heavy metals stimulate polyphosphate hydrolysis and the metal-phosphate complexes formed are transported out of the cell as part of a possibly functional heavy metal tolerance mechanism in A. ferrooxidans. PMID- 15345398 TI - Direct correlation between rates of anaerobic respiration and levels of mRNA for key respiratory genes in Geobacter sulfurreducens. AB - The predominance of Geobacter species in environments in which Fe(III) reduction is important has suggested that Fe(III) reduction rates might be estimated in Geobacter-dominated environments by assessing in situ activity with molecular techniques. To determine whether mRNA levels of key respiratory genes might be correlated with respiration rates in Geobacter sulfurreducens, studies were conducted with fumarate as the electron acceptor and acetate as the limiting electron donor in anaerobic continuous cultures. Levels of mRNA for a fumarate reductase gene, frdA, quantified by real-time reverse transcription-PCR were directly correlated with fumarate reduction rates. In similar studies with Fe(III) as the electron acceptor, mRNA levels for omcB, a gene for an outer membrane c-type cytochrome involved in Fe(III) reduction, were positively correlated with Fe(III) reduction rates. Levels of mRNA for frdA and omcB were also positively correlated with fumarate and Fe(III) reduction rates, respectively, when growth was limited by the availability of fumarate or Fe(III), but mRNA levels were higher than in acetate-limited cultures. Levels of mRNA for omcC, which encodes a c-type cytochrome highly similar to OmcB but not necessary for Fe(III) reduction, followed patterns different than those of omcB. This agrees with the previous finding that OmcC is not involved in Fe(III) reduction and suggests that changes in mRNA levels of omcB are related to its role in Fe(III) reduction. These results demonstrate that mRNA levels for respiratory genes might be used to estimate in situ Fe(III) reduction rates in Geobacter dominated environments but suggest that information on environmental conditions and/or the metabolic state of Geobacter species is also required for accurate rate estimates. PMID- 15345399 TI - Cell density-dependent gene contributes to efficient seed colonization by Pseudomonas putida KT2440. AB - We have characterized the expression pattern of a gene, ddcA, involved in initial colonization of corn seeds by Pseudomonas putida KT2440. The ddcA gene codes for a putative membrane polypeptide belonging to a family of conserved proteins of unknown function. Members of this family are widespread among prokaryotes and include the products of a Salmonella enterica serovar Typhimurium gene expressed during invasion of macrophages and psiE, an Escherichia coli phosphate starvation inducible gene. Although its specific role is undetermined, the presence of ddcA in multicopy restored the seed adhesion capacity of a KT2440 ddcA mutant. Expression of ddcA is growth phase regulated, being maximal at the beginning of stationary phase. It is independent of RpoS, nutrient depletion, or phosphate starvation, and it is not the result of changes in the medium pH during growth. Expression of ddcA is directly dependent on cell density, being also stimulated by the addition of conditioned medium and of seed exudates. This is the first evidence suggesting the existence of a quorum-sensing system in P. putida KT2440. The potential implication of such a signaling process in seed adhesion and colonization by the bacterium is discussed. PMID- 15345400 TI - Identification of specific in vivo-induced (ivi) genes in Yersinia ruckeri and analysis of ruckerbactin, a catecholate siderophore iron acquisition system. AB - This work reports the utilization of an in vivo expression technology system to identify in vivo-induced (ivi) genes in Yersinia ruckeri after determination of the conditions needed for its selection in fish. Fourteen clones were selected, and the cloned DNA fragments were analyzed after partial sequencing. In addition to sequences with no significant similarity, homology with genes encoding proteins putatively involved in two-component and type IV secretion systems, adherence, specific metabolic functions, and others were found. Among these sequences, four were involved in iron acquisition through a catechol siderophore (ruckerbactin). Thus, unlike other pathogenic yersiniae producing yersiniabactin, Y. ruckeri might be able to produce and utilize only this phenolate. The genetic organization of the ruckerbactin biosynthetic and uptake loci was similar to that of the Escherichia coli enterobactin gene cluster. Genes rucC and rupG, putative counterparts of E. coli entC and fepG, respectively, involved in the biosynthesis and transport of the iron siderophore complex, respectively, were analyzed further. Thus, regulation of expression by iron and temperature and their presence in other Y. ruckeri siderophore-producing strains were confirmed for these two loci. Moreover, 50% lethal dose values 100-fold higher than those of the wild-type strain were obtained with the rucC isogenic mutant, showing the importance of ruckerbactin in the pathogenesis caused by this microorganism. PMID- 15345401 TI - Acidified litter benefits the intestinal flora balance of broiler chickens. AB - The alterations in the balance of the normal intestinal bacterial flora of chickens exposed to acidified wood-derived litter were analyzed and compared to those of a control group exposed to nonacidified litter. A total of 1,728 broilers were divided into two groups, with six replicates in each. One group was exposed to dry wood-derived litter, and the other was exposed to dry wood-derived litter sprayed with a mixture of sodium lignosulfonate, formic acid, and propionic acid. At five different times, five chickens from each pen were killed and the intestinal contents from ileum and caeca were collected. The samples were diluted and plated onto selective media to identify coliforms, Lactobacillus spp., Clostridium perfringens, and Enterococcus spp. Covariance analysis of bacterial counts showed significantly lower counts for C. perfringens in the caeca and the ileum and for Enterococcus spp. and Lactobacillus spp. in the ileum in chickens exposed to the acidified litter. Lactobacillus spp. showed significantly higher counts in the caeca in chickens exposed to acidified litter. There was no difference between the two litters with regard to coliforms in the ileum and the caeca or to Enterococcus spp. in the caeca. The study shows that exposing the chickens to acidified litter lowers the intestinal bacterial number, especially in the ileum, without negative consequences for the chicken's health or performance. Of special interest are the lower counts of C. perfringens and Enterococcus spp. that might reduce the risk of developing clinical or subclinical necrotic enteritis and growth depression. PMID- 15345402 TI - Requirement of a relatively high threshold level of Mg(2+) for cell growth of a rhizoplane bacterium, Sphingomonas yanoikuyae EC-S001. AB - Mg(2+) is one of the essential elements for bacterial cell growth. The presence of the magnesium cation (Mg(2+)) in various concentrations often affects cell growth restoration in plant-associating bacteria. This study attempted to determine whether Mg(2+) levels in Sphingomonas yanoikuyae EC-S001 affected cell growth restoration in the host plant and what the threshold level is. S. yanoikuyae EC-S001, isolated from the rhizoplane of spinach seedlings grown from surface-sterilized seeds under aseptic conditions, displayed uniform dispersion and attachment throughout the rhizoplane and phylloplane of the host seedlings. S. yanoikuyae EC-S001 did not grow in potato-dextrose broth medium but grew well in an aqueous extract of spinach leaves. Chemical investigation of the growth factor in the spinach leaf extract led to identification of the active principle as the magnesium cation. A concentration of ca. 0.10 mM Mg(2+) or more allowed S. yanoikuyae EC-S001 to grow in potato-dextrose broth medium. Some saprophytic and/or diazotrophic bacteria used in our experiment were found to have diverse threshold levels for their Mg(2+) requirements. For example, Burkholderia cepacia EC-K014, originally isolated from the rhizoplane of a Melastoma sp., could grow even in Mg(2+)-free Hoagland's no. 2 medium with saccharose and glutamine (HSG medium) and requires a trace level of Mg(2+) for its growth. In contrast, S. yanoikuyae EC-S001, together with Bacillus subtilis IFO12113, showed the most drastic restoring responses to subsequent addition of 0.98 mM Mg(2+) to Mg(2+) free HSG medium. Our studies concluded that Mg(2+) is more than just the essential trace element needed for cell growth restoration in S. yanoikuyae EC S001 and that certain nonculturable bacteria may require a higher concentration of Mg(2+) or another specific essential element for their growth. PMID- 15345403 TI - In situ monitoring of streptothricin production by Streptomyces rochei F20 in soil and rhizosphere. AB - The onset of streptothricin (ST) biosynthesis in Streptomyces rochei F20 was studied by using reverse transcription-PCR (RT-PCR) to detect transcripts of ST genes during growth in liquid medium, soil, and the rhizosphere. In situ results correlated with those obtained in vitro, illustrating the growth phase-dependent manner of ST production by F20. Maximal transcription of ST resistance (sttR) and biosynthesis (sttA) genes occurred during the transition between the exponential and stationary phases of growth, when the specific growth rate (micro) started to decline. A higher level of gene expression of sttR versus sttA was observed in all experiments. In liquid culture, maximal transcript accumulation of the sttA gene was only ca. 40% that of the sttR gene. sttA and sttR mRNAs were detected in soil containing approximately 10(6) CFU of growing cells g of soil(-1). sttR mRNA was detected in sterile and nonsterile rhizosphere colonized with growing mycelium of F20 at 1.2 x 10(6) and 4.0 x 10(5) CFU g of soil(-1), respectively. However, neither sttR nor sttA transcripts were detected by RT-PCR in the rhizoplane, which supported a lower population density of F20 than the rhizosphere. PMID- 15345404 TI - Nonmarine crenarchaeol in Nevada hot springs. AB - Glycerol dialkyl glycerol tetraethers (GDGTs) are core membrane lipids of the Crenarchaeota. The structurally unusual GDGT crenarchaeol has been proposed as a taxonomically specific biomarker for the marine planktonic group I archaea. It is found ubiquitously in the marine water column and in sediments. In this work, samples of microbial community biomass were obtained from several alkaline and neutral-pH hot springs in Nevada, United States. Lipid extracts of these samples were analyzed by high-performance liquid chromatography-mass spectrometry and by gas chromatography-mass spectrometry. Each sample contained GDGTs, and among these compounds was crenarchaeol. The distribution of archaeal lipids in Nevada hot springs did not appear to correlate with temperature, as has been observed in the marine environment. Instead, a significant correlation with the concentration of bicarbonate was observed. Archaeal DNA was analyzed by denaturing gradient gel electrophoresis. All samples contained 16S rRNA gene sequences which were more strongly related to thermophilic crenarchaeota than to Cenarchaeum symbiosum, a marine nonthermophilic crenarchaeon. The occurrence of crenarchaeol in environments containing sequences affiliated with thermophilic crenarchaeota suggests a wide phenotypic distribution of this compound. The results also indicate that crenarchaeol can no longer be considered an exclusive biomarker for marine species. PMID- 15345405 TI - Substrate-induced production and secretion of cellulases by Clostridium acetobutylicum. AB - Clostridium acetobutylicum ATCC 824 is a solventogenic bacterium that grows heterotrophically on a variety of carbohydrates, including glucose, cellobiose, xylose, and lichenan, a linear polymer of beta-1,3- and beta-1,4-linked beta-D glucose units. C. acetobutylicum does not degrade cellulose, although its genome sequence contains several cellulase-encoding genes and a complete cellulosome cluster of cellulosome genes. In the present study, we demonstrate that a low but significant level of induction of cellulase activity occurs during growth on xylose or lichenan. The celF gene, located in the cellulosome-like gene cluster and coding for a unique cellulase that belongs to glycoside hydrolase family 48, was cloned in Escherichia coli, and antibodies were raised against the overproduced CelF protein. A Western blot analysis suggested a possible catabolite repression by glucose or cellobiose and an up-regulation by lichenan or xylose of the extracellular production of CelF by C. acetobutylicum. Possible reasons for the apparent inability of C. acetobutylicum to degrade cellulose are discussed. PMID- 15345406 TI - Molecular characterization of CcpA and involvement of this protein in transcriptional regulation of lactate dehydrogenase and pyruvate formate-lyase in the ruminal bacterium Streptococcus bovis. AB - A ccpA gene that encodes global catabolite control protein A (CcpA) in Streptococcus bovis was identified and characterized, and the involvement of CcpA in transcriptional control of a gene (ldh) encoding lactate dehydrogenase (LDH) and a gene (pfl) encoding pyruvate formate-lyase (PFL) was examined. The ccpA gene was shown to be transcribed as a monocistronic operon. A catabolite responsive element (cre) was found in the promoter region of ccpA, suggesting that ccpA transcription in S. bovis is autogenously regulated. CcpA required HPr that was phosphorylated at the serine residue at position 46 (HPr-[Ser-P]) for binding to the cre site, but glucose 6-phosphate, fructose 1,6-bisphosphate, and NADP had no effect on binding. Diauxic growth was observed when S. bovis was grown in a medium containing glucose and lactose, but it disappeared when ccpA was disrupted, which indicates that CcpA is involved in catabolite repression in S. bovis. The level of ccpA mRNA was higher when cells were grown on glucose than when they were grown on lactose, which was in line with the level of ldh mRNA. When cells were grown on glucose, the ldh mRNA level was lower but the pfl mRNA level was higher in a ccpA-disrupted mutant than in the parent strain, which suggests that ldh transcription is enhanced and pfl transcription is suppressed by CcpA. The ccpA-disrupted mutant produced less lactate and more formate than the parent, probably because the mutant had reduced LDH activity and elevated PFL activity. In the upper region of both ldh and pfl, a cre-like sequence was found, suggesting that the complex consisting of CcpA and HPr-[Ser-P] binds to the possible cre sites. Thus, CcpA appears to be involved in the global regulation of sugar utilization in S. bovis. PMID- 15345407 TI - Anaerobic growth of Bacillus mojavensis and Bacillus subtilis requires deoxyribonucleosides or DNA. AB - Bacillus mojavensis strains JF-2 (ATCC 39307), ROB2, and ABO21191(T) and Bacillus subtilis strains 168 (ATCC 23857) and ATCC 12332 required four deoxyribonucleosides or DNA for growth under strict anaerobic conditions. Bacillus licheniformis strains L89-11 and L87-11, Bacillus sonorensis strain TG8 8, and Bacillus cereus (ATCC 14579) did not require DNA for anaerobic growth. The requirement for the deoxyribonucleosides or DNA did not occur under aerobic growth conditions. The addition of a mixture of five nucleic acid bases, four ribonucleotides, or four ribonucleosides to the basal medium did not replace the requirement of B. mojavensis JF-2 for the four deoxyribonucleosides. However, the addition of salmon sperm DNA, herring sperm DNA, Escherichia coli DNA, or synthetic DNA (single or double stranded) to the basal medium supported anaerobic growth. The addition of four deoxyribonucleosides to the basal medium allowed aerobic growth of B. mojavensis JF-2 in the presence of hydroxyurea. B. mojavensis did not grow in DNA-supplemented basal medium that lacked sucrose as the energy source. These data provide strong evidence that externally supplied deoxyribonucleosides can be used to maintain a balanced deoxyribonucleotide pool for DNA synthesis and suggest that ribonucleotide reductases may not be essential to the bacterial cell cycle nor are they necessarily part of a minimal bacterial genome. PMID- 15345408 TI - Combined use of cultivation-dependent and cultivation-independent methods indicates that members of most haloarchaeal groups in an Australian crystallizer pond are cultivable. AB - Haloarchaea are the dominant microbial flora in hypersaline waters with near saturating salt levels. The haloarchaeal diversity of an Australian saltern crystallizer pond was examined by use of a library of PCR-amplified 16S rRNA genes and by cultivation. High viable counts (10(6) CFU/ml) were obtained on solid media. Long incubation times (> or =8 weeks) appeared to be more important than the medium composition for maximizing viable counts and diversity. Of 66 isolates examined, all belonged to the family Halobacteriaceae, including members related to species of the genera Haloferax, Halorubrum, and Natronomonas. In addition, isolates belonging to a novel group (the ADL group), previously detected only as 16S rRNA genes in an Antarctic hypersaline lake (Deep Lake), were cultivated for the first time. The 16S rRNA gene library identified the following five main groups: Halorubrum groups 1 and 2 (49%), the SHOW (square haloarchaea of Walsby) group (33%), the ADL group (16%), and the Natronomonas group (2%). There were two significant differences between the organisms detected in cultivation and 16S rRNA sequence results. Firstly, Haloferax spp. were frequently isolated on plates (15% of all isolates) but were not detected in the 16S rRNA sequences. Control experiments indicated that a bias against Haloferax sequences in the generation of the 16S rRNA gene library was unlikely, suggesting that Haloferax spp. readily form colonies, even though they were not a dominant group. Secondly, while the 16S rRNA gene library identified the SHOW group as a major component of the microbial community, no isolates of this group were obtained. This inability to culture members of the SHOW group remains an outstanding problem in studying the ecology of hypersaline environments. PMID- 15345409 TI - Contribution of fungi and bacteria to leaf litter decomposition in a polluted river. AB - The contribution of fungi and bacteria to the decomposition of alder leaves was examined at two reference and two polluted sites in the Ave River (northwestern Portugal). Leaf mass loss, microbial production from incorporation rates of radiolabeled compounds into biomolecules, fungal biomass from ergosterol concentration, sporulation rates, and diversity of aquatic hyphomycetes associated with decomposing leaves were determined. The concentrations of organic nutrients and of inorganic nitrogen and phosphorus in the stream water was elevated and increased at downstream sites. Leaf decomposition rates were high (0.013 day(-1) < k < 0.042 day(-1)), and the highest value was estimated at the most downstream polluted site, where maximum values of microbial production and fungal biomass and sporulation were found. The slowest decomposition occurred at the other polluted site, where, along with the nutrient enrichment, the lowest current velocity and dissolved-oxygen concentration in water were observed. At this site, fungal production, biomass, and sporulation were depressed, suggesting that stimulation of fungal activity by increased nutrient concentrations might be offset by other factors. Although bacterial production was higher at polluted sites, fungi accounted for more than 94% of the total microbial net production. Fungal yield coefficients varied from 10.2 to 13.6%, while those of bacteria were less than 1%. The contribution of fungi to overall leaf carbon loss (29.0 to 38.8%) greatly exceeded that of bacteria (4.2 to 13.9%). PMID- 15345410 TI - Comparative proteomic analysis of extracellular proteins of enterohemorrhagic and enteropathogenic Escherichia coli strains and their ihf and ler mutants. AB - Enterohemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC, respectively) strains are closely related human pathogens that are responsible for food-borne epidemics in many countries. Integration host factor (IHF) and the locus of enterocyte effacement-encoded regulator (Ler) are needed for the expression of virulence genes in EHEC and EPEC, including the elicitation of actin rearrangements for attaching and effacing lesions. We applied a proteomic approach, using two-dimensional polyacrylamide gel electrophoresis in combination with matrix-assisted laser desorption ionization-time of flight mass spectrometry and a protein database search, to analyze the extracellular protein profiles of EHEC EDL933, EPEC E2348/69, and their ihf and ler mutants. Fifty-nine major protein spots from the extracellular proteomes were identified, including six proteins of unknown function. Twenty-six of them were conserved between EHEC EDL933 and EPEC E2348/69, while some of them were strain-specific proteins. Four common extracellular proteins (EspA, EspB, EspD, and Tir) were regulated by both IHF and Ler in EHEC EDL933 and EPEC E2348/69. TagA in EHEC EDL933 and EspC and EspF in EPEC E2348/69 were present in the wild-type strains but absent from their respective ler and ihf mutants, while FliC was overexpressed in the ihf mutant of EPEC E2348/69. Two dominant forms of EspB were found in EHEC EDL933 and EPEC E2348/69, but the significance of this is unknown. These results show that proteomics is a powerful platform technology for accelerating the understanding of EPEC and EHEC pathogenesis and identifying markers for laboratory diagnoses of these pathogens. PMID- 15345411 TI - Degradation of estrogens by Rhodococcus zopfii and Rhodococcus equi isolates from activated sludge in wastewater treatment plants. AB - We have isolated four strains of Rhodococcus which specifically degrade estrogens by using enrichment culture of activated sludge from wastewater treatment plants. Strain Y 50158, identified as Rhodococcus zopfii, completely and rapidly degraded 100 mg of 17beta-estradiol, estrone, estriol, and ethinyl estradiol/liter, as demonstrated by thin-layer chromatography and gas chromatography-mass spectrometry analyses. Strains Y 50155, Y 50156, and Y 50157, identified as Rhodococcus equi, showed degradation activities comparable with that of Y 50158. Using the random amplified polymorphism DNA fingerprinting test, these three strains were confirmed to have been derived from different sources. R. zopfii Y 50158, which showed the highest activity among these four strains, revealed that the strain selectively degraded 17beta-estradiol during jar fermentation, even when glucose was used as a readily utilizable carbon source in the culture medium. Measurement of estrogenic activities with human breast cancer-derived MVLN cells showed that these four strains each degraded 100 mg of 17beta estradiol/liter to 1/100 of the specific activity level after 24 h. It is thus suggested that these strains degrade 17beta-estradiol into substances without estrogenic activity. PMID- 15345412 TI - Multiple nonidentical reductive-dehalogenase-homologous genes are common in Dehalococcoides. AB - Degenerate primers were used to amplify large fragments of reductive-dehalogenase homologous (RDH) genes from genomic DNA of two Dehalococcoides populations, the chlorobenzene- and dioxin-dechlorinating strain CBDB1 and the trichloroethene dechlorinating strain FL2. The amplicons (1,350 to 1,495 bp) corresponded to nearly complete open reading frames of known reductive dehalogenase genes and short fragments (approximately 90 bp) of genes encoding putative membrane anchoring proteins. Cloning and restriction analysis revealed the presence of at least 14 different RDH genes in each strain. All amplified RDH genes showed sequence similarity with known reductive dehalogenase genes over the whole length of the sequence and shared all characteristics described for reductive dehalogenases. Deduced amino acid sequences of seven RDH genes from strain CBDB1 were 98.5 to 100% identical to seven different RDH genes from strain FL2, suggesting that both strains have an overlapping substrate range. All RDH genes identified in strains CBDB1 and FL2 were related to the RDH genes present in the genomes of Dehalococcoides ethenogenes strain 195 and Dehalococcoides sp. strain BAV1; however, sequence identity did not exceed 94.4 and 93.1%, respectively. The presence of RDH genes in strains CBDB1, FL2, and BAV1 that have no orthologs in strain 195 suggests that these strains possess dechlorination activities not present in strain 195. Comparative sequence analysis identified consensus sequences for cobalamin binding in deduced amino acid sequences of seven RDH genes. In conclusion, this study demonstrates that the presence of multiple nonidentical RDH genes is characteristic of Dehalococcoides strains. PMID- 15345413 TI - Modified serial analysis of gene expression method for construction of gene expression profiles of microbial eukaryotic species. AB - Serial analysis of gene expression (SAGE) is a powerful approach for the identification of differentially expressed genes, providing comprehensive and quantitative gene expression profiles in the form of short tag sequences. Each tag represents a unique transcript, and the relative frequencies of tags in the SAGE library are equal to the relative proportions of the transcripts they represent. One of the major obstacles in the preparation of SAGE libraries from microorganisms is the requirement for large amounts of starting material (i.e., mRNA). Here, we present a novel approach for the construction of SAGE libraries from small quantities of total RNA by using Y linkers to selectively amplify 3' cDNA fragments. To validate this method, we constructed comprehensive gene expression profiles of the toxic dinoflagellate Pfiesteria shumwayae. SAGE libraries were constructed from an actively toxic fish-fed culture of P. shumwayae and from a recently toxic alga-fed culture. P. shumwayae-specific gene transcripts were identified by comparison of tag sequences in the two libraries. Representative tags with frequencies ranging from 0.026 to 3.3% of the total number of tags in the libraries were chosen for further analysis. Expression of each transcript was confirmed in separate control cultures of toxic P. shumwayae. The modified SAGE method described here produces gene expression profiles that appear to be both comprehensive and quantitative, and it is directly applicable to the study of gene expression in other environmentally relevant microbial species. PMID- 15345414 TI - Phenotypic and molecular typing of Salmonella strains reveals different contamination sources in two commercial pig slaughterhouses. AB - This study aimed to define the origin of Salmonella contamination on swine carcasses and the distribution of Salmonella serotypes in two commercial slaughterhouses during normal activity. Salmonellae were isolated from carcasses, from colons and mesenteric lymph nodes of individual pigs, and from the slaughterhouse environment. All strains were serotyped; Salmonella enterica serotype Typhimurium and Salmonella enterica serotype Derby isolates were additionally typed beyond the serotype level by pulsed-field gel electrophoresis (PFGE) and antibiotic resistance profiling (ARP); and a subset of 31 serotype Typhimurium strains were additionally phage typed. PFGE and ARP had the same discriminative possibility. Phage typing in combination with PFGE could give extra information for some strains. In one slaughterhouse, 21% of the carcasses were contaminated, reflecting a correlation with the delivery of infected pigs. Carcass contamination did not result only from infection of the corresponding pig; only 25% of the positive carcasses were contaminated with the same serotype or genotype found in the corresponding feces or mesenteric lymph nodes. In the other slaughterhouse, 70% of the carcasses were contaminated, and only in 4% was the same genotype or serotype detected as in the feces of the corresponding pigs. The other positive carcasses in both slaughterhouses were contaminated by genotypes present in the feces or lymph nodes of pigs slaughtered earlier that day or from dispersed sources in the environment. In slaughterhouses, complex contamination cycles may be present, resulting in the isolation of many different genotypes circulating in the environment due to the supply of positive animals and in the contamination of carcasses, probably through aerosols. PMID- 15345415 TI - Identification and inactivation of genetic loci involved with Lactobacillus acidophilus acid tolerance. AB - Amino acid decarboxylation-antiporter reactions are one of the most important systems for maintaining intracellular pH between physiological limits under acid stress. We analyzed the Lactobacillus acidophilus NCFM complete genome sequence and selected four open reading frames with similarities to genes involved with decarboxylation reactions involved in acid tolerance in several microorganisms. Putative genes encoding an ornithine decarboxylase, an amino acid permease, a glutamate gamma-aminobutyrate antiporter, and a transcriptional regulator were disrupted by insertional inactivation. The ability of L. acidophilus to survive low-pH conditions, such as those encountered in the stomach or fermented dairy foods, was investigated and compared to the abilities of early- and late stationary-phase cells of the mutants by challenging them with a variety of acidic conditions. All of the integrants were more sensitive to low pH than the parental strain. Interestingly, each integrant also exhibited an adaptive acid response during logarithmic growth, indicating that multiple mechanisms are present and orchestrated in L. acidophilus in response to acid challenge. PMID- 15345416 TI - Role of hexose transport in control of glycolytic flux in Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae predominantly ferments glucose to ethanol at high external glucose concentrations, irrespective of the presence of oxygen. In contrast, at low external glucose concentrations and in the presence of oxygen, as in a glucose-limited chemostat, no ethanol is produced. The importance of the external glucose concentration suggests a central role for the affinity and maximal transport rates of yeast's glucose transporters in the control of ethanol production. Here we present a series of strains producing functional chimeras between the hexose transporters Hxt1 and Hxt7, each of which has distinct glucose transport characteristics. The strains display a range of decreasing glycolytic rates resulting in a proportional decrease in ethanol production. Using these strains, we show for the first time that at high glucose levels, the glucose uptake capacity of wild-type S. cerevisiae does not control glycolytic flux during exponential batch growth. In contrast, our chimeric Hxt transporters control the rate of glycolysis to a high degree. Strains whose glucose uptake is mediated by these chimeric transporters will undoubtedly provide a powerful tool with which to examine in detail the mechanism underlying the switch between fermentation and respiration in S. cerevisiae and will provide new tools for the control of industrial fermentations. PMID- 15345417 TI - Transmission and infectious dose of Escherichia coli O157:H7 in swine. AB - Escherichia coli O157:H7 is only occasionally isolated from healthy swine, but some experimentally infected animals will shed the organism in their feces for at least 2 months. Potential explanations for the paucity of naturally occurring infections in swine, as compared to cattle, include a lack of animal-to-animal transmission so that the organism cannot be maintained within a herd, a high infectious dose, or herd management practices that prevent the maintenance of the organism in the gastrointestinal tract. We hypothesized that donor pigs infected with E. coli O157:H7 would transmit the organism to naive pigs. We also determined the infectious dose and whether housing pigs individually on grated floors would decrease the magnitude or duration of fecal shedding. Infected donor pigs shedding <10(4) CFU of E. coli O157:H7 per g transmitted the organism to 6 of 12 naive pigs exposed to them. The infectious dose of E. coli O157:H7 for 3 month-old pigs was approximately 6 x 10(3) CFU. There was no difference in the magnitude and duration of fecal shedding by pigs housed individually on grates compared to those housed two per pen on cement floors. These results suggest that swine do not have an innate resistance to colonization by E. coli O157:H7 and that they could serve as a reservoir host under suitable conditions. PMID- 15345418 TI - Effect of forage or grain diets with or without monensin on ruminal persistence and fecal Escherichia coli O157:H7 in cattle. AB - Twelve ruminally cannulated cattle, adapted to forage or grain diet with or without monensin, were used to investigate the effects of diet and monensin on concentration and duration of ruminal persistence and fecal shedding of E. coli O157:H7. Cattle were ruminally inoculated with a strain of E. coli O157:H7 (10(10) CFU/animal) made resistant to nalidixic acid (Nal(r)). Ruminal and fecal samples were collected for 11 weeks, and then cattle were euthanized and necropsied and digesta from different gut locations were collected. Samples were cultured for detection and enumeration of Nal(r) E. coli O157:H7. Cattle fed forage diets were culture positive for E. coli O157:H7 in the feces for longer duration (P < 0.05) than cattle fed a grain diet. In forage-fed cattle, the duration they remained culture positive for E. coli O157:H7 was shorter (P < 0.05) when the diet included monensin. Generally, ruminal persistence of Nal(r) E. coli O157:H7 was not affected by diet or monensin. At necropsy, E. coli O157:H7 was detected in cecal and colonic digesta but not from the rumen. Our study showed that cattle fed a forage diet were culture positive longer and with higher numbers than cattle on a grain diet. Monensin supplementation decreased the duration of shedding with forage diet, and the cecum and colon were culture positive for E. coli O157:H7 more often than the rumen of cattle. PMID- 15345419 TI - Comparison of fluorescence microscopy and solid-phase cytometry methods for counting bacteria in water. AB - Total direct counts of bacterial abundance are central in assessing the biomass and bacteriological quality of water in ecological and industrial applications. Several factors have been identified that contribute to the variability in bacterial abundance counts when using fluorescent microscopy, the most significant of which is retaining an adequate number of cells per filter to ensure an acceptable level of statistical confidence in the resulting data. Previous studies that have assessed the components of total-direct-count methods that contribute to this variance have attempted to maintain a bacterial cell abundance value per filter of approximately 10(6) cells filter(-1). In this study we have established the lower limit for the number of bacterial cells per filter at which the statistical reliability of the abundance estimate is no longer acceptable. Our results indicate that when the numbers of bacterial cells per filter were progressively reduced below 10(5), the microscopic methods increasingly overestimated the true bacterial abundance (range, 15.0 to 99.3%). The solid-phase cytometer only slightly overestimated the true bacterial abundances and was more consistent over the same range of bacterial abundances per filter (range, 8.9 to 12.5%). The solid-phase cytometer method for conducting total direct counts of bacteria was less biased and performed significantly better than any of the microscope methods. It was also found that microscopic count data from counting 5 fields on three separate filters were statistically equivalent to data from counting 20 fields on a single filter. PMID- 15345420 TI - RsbT and RsbV contribute to sigmaB-dependent survival under environmental, energy, and intracellular stress conditions in Listeria monocytogenes. AB - Sigma B (sigma(B)) is a stress-responsive alternative sigma factor that has been identified in various gram-positive bacteria. Seven different regulators of sigma B (Rsbs) are located in the sigB operons of both Bacillus subtilis and Listeria monocytogenes. In B. subtilis, these proteins contribute to regulation of sigma(B) activity by conveying environmental and energy stress signals through two well-established branches of a signal transduction pathway. RsbT contributes to regulation of sigma(B) activity in response to environmental stresses, while RsbV contributes to sigma(B) activation under both environmental and energy stresses in B. subtilis. To probe L. monocytogenes Rsb roles in sigma(B)-mediated responses to various stresses, in-frame deletions were created in rsbT and rsbV. Phenotypic characterization of the L. monocytogenes rsbT and rsbV null mutants revealed that both mutants were similar to the DeltasigB strain in their abilities to survive under environmental stress conditions (exposure to synthetic gastric fluid, pH 2.5, acidified brain heart infusion broth [BHI], or oxidative stress [13 mM cumene hydroperoxide]). Under energy stress conditions (carbon starvation in defined media, entry into stationary phase, or reduced intracellular ATP), both DeltarsbT and DeltarsbV showed survival reductions similar to that of the DeltasigB strain. These observations suggest that the pathways for Rsb-dependent regulation of sigma(B) activity differ between L. monocytogenes and B. subtilis. As sigma(B) also activates transcription of the L. monocytogenes prfAP2 promoter, we evaluated virulence-associated characteristics of DeltaprfAP1rsbT and DeltaprfAP1rsbV double mutants in hemolysis and tissue culture assays. Both double mutants showed identical phenotypes to DeltaprfAP1P2 and DeltaprfAP1sigB double mutants, i.e., reduced hemolysis activity and reduced plaque size in mouse fibroblast cells. These findings indicate that RsbT and RsbV both contribute to sigma(B) activation in L. monocytogenes during exposure to environmental and energy stresses as well as during tissue culture infection. PMID- 15345421 TI - Localization and characterization of two novel genes encoding stereospecific dioxygenases catalyzing 2(2,4-dichlorophenoxy)propionate cleavage in Delftia acidovorans MC1. AB - Two novel genes, rdpA and sdpA, encoding the enantiospecific alpha-ketoglutarate dependent dioxygenases catalyzing R,S-dichlorprop cleavage in Delftia acidovorans MC1 were identified. Significant similarities to other known genes were not detected, but their deduced amino acid sequences were similar to those of other alpha-ketoglutarate dioxygenases. RdpA showed 35% identity with TauD of Pseudomonas aeruginosa, and SdpA showed 37% identity with TfdA of Ralstonia eutropha JMP134. The functionally important amino acid sequence motif HX(D/E)X(23 26)(T/S)X(114-183)HX(10-13)R/K, which is highly conserved in group II alpha ketoglutarate-dependent dioxygenases, was present in both dichlorprop-cleaving enzymes. Transposon mutagenesis of rdpA inactivated R-dichlorprop cleavage, indicating that it was a single-copy gene. Both rdpA and sdpA were located on the plasmid pMC1 that also carries the lower pathway genes. Sequencing of a 25.8-kb fragment showed that the dioxygenase genes were separated by a 13.6-kb region mainly comprising a Tn501-like transposon. Furthermore, two copies of a sequence similar to IS91-like elements were identified. Hybridization studies comparing the wild-type plasmid and that of the mutant unable to cleave dichlorprop showed that rdpA and sdpA were deleted, whereas the lower pathway genes were unaffected, and that deletion may be caused by genetic rearrangements of the IS91-like elements. Two other dichlorprop-degrading bacterial strains, Rhodoferax sp. strain P230 and Sphingobium herbicidovorans MH, were shown to carry rdpA genes of high similarity to rdpA from strain MC1, but sdpA was not detected. This suggested that rdpA gene products are involved in the degradation of R dichlorprop in these strains. PMID- 15345422 TI - Heads or tails: host-parasite interactions in the Drosophila-Wolbachia system. AB - Wolbachia strains are endosymbiotic bacteria typically found in the reproductive tracts of arthropods. These bacteria manipulate host reproduction to ensure maternal transmission. They are usually transmitted vertically, so it has been predicted that they have evolved a mechanism to target the host's germ cells during development. Through cytological analysis we found that Wolbachia strains display various affinities for the germ line of Drosophila. Different Wolbachia strains show posterior, anterior, or cortical localization in Drosophila embryos, and this localization is congruent with the classification of the organisms based on the wsp (Wolbachia surface protein) gene sequence. This embryonic distribution pattern is established during early oogenesis and does not change until late stages of embryogenesis. The posterior and anterior localization of Wolbachia resembles that of oskar and bicoid mRNAs, respectively, which define the anterior posterior axis in the Drosophila oocyte. By comparing the properties of a single Wolbachia strain in different host backgrounds and the properties of different Wolbachia strains in the same host background, we concluded that bacterial factors determine distribution, while bacterial density seems to be limited by the host. Possible implications concerning cytoplasmic incompatibility and evolution of strains are discussed. PMID- 15345423 TI - Biofuel cells select for microbial consortia that self-mediate electron transfer. AB - Microbial fuel cells hold great promise as a sustainable biotechnological solution to future energy needs. Current efforts to improve the efficiency of such fuel cells are limited by the lack of knowledge about the microbial ecology of these systems. The purposes of this study were (i) to elucidate whether a bacterial community, either suspended or attached to an electrode, can evolve in a microbial fuel cell to bring about higher power output, and (ii) to identify species responsible for the electricity generation. Enrichment by repeated transfer of a bacterial consortium harvested from the anode compartment of a biofuel cell in which glucose was used increased the output from an initial level of 0.6 W m(-2) of electrode surface to a maximal level of 4.31 W m(-2) (664 mV, 30.9 mA) when plain graphite electrodes were used. This result was obtained with an average loading rate of 1 g of glucose liter(-1) day(-1) and corresponded to 81% efficiency for electron transfer from glucose to electricity. Cyclic voltammetry indicated that the enhanced microbial consortium had either membrane bound or excreted redox components that were not initially detected in the community. Dominant species of the enhanced culture were identified by denaturing gradient gel electrophoresis and culturing. The community consisted mainly of facultative anaerobic bacteria, such as Alcaligenes faecalis and Enterococcus gallinarum, which are capable of hydrogen production. Pseudomonas aeruginosa and other Pseudomonas species were also isolated. For several isolates, electrochemical activity was mainly due to excreted redox mediators, and one of these mediators, pyocyanin produced by P. aeruginosa, could be characterized. Overall, the enrichment procedure, irrespective of whether only attached or suspended bacteria were examined, selected for organisms capable of mediating the electron transfer either by direct bacterial transfer or by excretion of redox components. PMID- 15345424 TI - Microautoradiographic study of Rhodocyclus-related polyphosphate-accumulating bacteria in full-scale enhanced biological phosphorus removal plants. AB - The ecophysiology of uncultured Rhodocyclus-related polyphosphate-accumulating organisms (PAO) present in three full-scale enhanced biological phosphorus removal (EBPR) activated sludge plants was studied by using microautoradiography combined with fluorescence in situ hybridization. The investigations showed that these organisms were present in all plants examined and constituted 5 to 10, 10 to 15, and 17 to 22% of the community biomass. The behavior of these bacteria generally was consistent with the biochemical models proposed for PAO, based on studies of lab-scale investigations of enriched and often unknown PAO cultures. Rhodocyclus-related PAO were able to accumulate short-chain substrates, including acetate, propionate, and pyruvate, under anaerobic conditions, but they could not assimilate many other low-molecular-weight compounds, such as ethanol and butyrate. They were able to assimilate two substrates (e.g., acetate and propionate) simultaneously. Leucine and thymidine could not be assimilated as sole substrates and could only be assimilated as cosubstrates with acetate, perhaps serving as N sources. Glucose could not be assimilated by the Rhodocyclus related PAO, but it was easily fermented in the sludge to products that were subsequently consumed. Glycolysis, and not the tricarboxylic acid cycle, was the source that provided the reducing power needed by the Rhodocyclus-related PAO to form the intracellular polyhydroxyalkanoate storage compounds during anaerobic substrate assimilation. The Rhodocyclus-related PAO were able to take up orthophosphate and accumulate polyphosphate when oxygen, nitrate, or nitrite was present as an electron acceptor. Furthermore, in the presence of acetate growth was sustained by using oxygen, as well as nitrate or nitrite, as an electron acceptor. This strongly indicates that Rhodocyclus-related PAO were able to denitrify and thus played a role in the denitrification occurring in full-scale EBPR plants. PMID- 15345425 TI - Epiphytic cyanobacteria on Chara vulgaris are the main contributors to N(2) fixation in rice fields. AB - The distribution of nitrogenase activity in the rice-soil system and the possible contribution of epiphytic cyanobacteria on rice plants and other macrophytes to this activity were studied in two locations in the rice fields of Valencia, Spain, in two consecutive crop seasons. The largest proportion of photodependent N(2) fixation was associated with the macrophyte Chara vulgaris in both years and at both locations. The nitrogen fixation rate associated with Chara always represented more than 45% of the global nitrogenase activity measured in the rice field. The estimated average N(2) fixation rate associated with Chara was 27.53 kg of N ha(-1) crop(-1). The mean estimated N(2) fixation rates for the other parts of the system for all sampling periods were as follows: soil, 4.07 kg of N ha(-1) crop(-1); submerged parts of rice plants, 3.93 kg of N ha(-1) crop(-1); and roots, 0.28 kg of N ha(-1) crop(-1). Micrographic studies revealed the presence of epiphytic cyanobacteria on the surface of Chara. Three-dimensional reconstructions by confocal scanning laser microscopy revealed no cyanobacterial cells inside the Chara structures. Quantification of epiphytic cyanobacteria by image analysis revealed that cyanobacteria were more abundant in nodes than in internodes (on average, cyanobacteria covered 8.4% +/- 4.4% and 6.2% +/- 5.0% of the surface area in the nodes and internodes, respectively). Epiphytic cyanobacteria were also quantified by using a fluorometer. This made it possible to discriminate which algal groups were the source of chlorophyll a. Chlorophyll a measurements confirmed that cyanobacteria were more abundant in nodes than in internodes (on average, the chlorophyll a concentrations were 17.2 +/- 28.0 and 4.0 +/- 3.8 microg mg [dry weight] of Chara(-1) in the nodes and internodes, respectively). These results indicate that this macrophyte, which is usually considered a weed in the context of rice cultivation, may help maintain soil N fertility in the rice field ecosystem. PMID- 15345426 TI - New expression system tightly controlled by zinc availability in Lactococcus lactis. AB - Here we developed the new expression system P(Zn) zitR, based on the regulatory signals (P(Zn) promoter and zitR repressor) of the Lactococcus lactis zit operon, involved in Zn(2+) high-affinity uptake and regulation. A P(Zn) zitR-controlled expression vector was constructed, and expression regulation was studied with two reporter genes, uspnuc and lacLM; these genes encode, respectively, a protein derived from Staphylococcus aureus secreted nuclease and Leuconostoc mesenteroides cytoplasmic beta-galactosidase. Nuclease and beta-galactosidase activities of L. lactis MG1363 cells expressing either uspnuc or lacLM under the control of P(Zn) zitR were evaluated on plates and quantified from liquid cultures as a function of divalent metal ion, particularly Zn(2+), availability in the environment. Our results demonstrate that P(Zn) zitR is highly inducible upon divalent cation starvation, obtained either through EDTA addition or during growth in chemically defined medium, and is strongly repressed in the presence of excess Zn(2+). The efficiency of the P(Zn) zitR expression system was compared to that of the well-known nisin-controlled expression (NICE) system with the same reporter genes cloned under either P(Zn) zitR or P(nisA) nisRK control. lacLM induction levels reached with both systems were on the same order of magnitude, even though the NICE system is fivefold more efficient than the P(Zn) zitR system. An even smaller difference or no difference was observed after 3 h of induction when nuclease was used as a reporter for Western blotting detection. P(Zn) zitR proved to be a powerful expression system for L. lactis, as it is tightly controlled by the zinc concentration in the medium. PMID- 15345427 TI - Construction of a xylan-fermenting yeast strain through codisplay of xylanolytic enzymes on the surface of xylose-utilizing Saccharomyces cerevisiae cells. AB - Hemicellulose is one of the major forms of biomass in lignocellulose, and its essential component is xylan. We used a cell surface engineering system based on alpha-agglutinin to construct a Saccharomyces cerevisiae yeast strain codisplaying two types of xylan-degrading enzymes, namely, xylanase II (XYNII) from Trichoderma reesei QM9414 and beta-xylosidase (XylA) from Aspergillus oryzae NiaD300, on the cell surface. In a high-performance liquid chromatography analysis, xylose was detected as the main product of the yeast strain codisplaying XYNII and XylA, while xylobiose and xylotriose were detected as the main products of a yeast strain displaying XYNII on the cell surface. These results indicate that xylan is sequentially hydrolyzed to xylose by the codisplayed XYNII and XylA. In a further step toward achieving the simultaneous saccharification and fermentation of xylan, a xylan-utilizing S. cerevisiae strain was constructed by codisplaying XYNII and XylA and introducing genes for xylose utilization, namely, those encoding xylose reductase and xylitol dehydrogenase from Pichia stipitis and xylulokinase from S. cerevisiae. After 62 h of fermentation, 7.1 g of ethanol per liter was directly produced from birchwood xylan, and the yield in terms of grams of ethanol per gram of carbohydrate consumed was 0.30 g/g. These results demonstrate that the direct conversion of xylan to ethanol is accomplished by the xylan-utilizing S. cerevisiae strain. PMID- 15345428 TI - Shewanella oneidensis MR-1 restores menaquinone synthesis to a menaquinone negative mutant. AB - The mechanisms underlying the use of insoluble electron acceptors by metal reducing bacteria, such as Shewanella oneidensis MR-1, are currently under intensive study. Current models for shuttling electrons across the outer membrane (OM) of MR-1 include roles for OM cytochromes and the possible excretion of a redox shuttle. While MR-1 is able to release a substance that restores the ability of a menaquinone (MK)-negative mutant, CMA-1, to reduce the humic acid analog anthraquinone-2,6-disulfonate (AQDS), cross-feeding experiments conducted here showed that the substance released by MR-1 restores the growth of CMA-1 on several soluble electron acceptors. Various strains derived from MR-1 also release this substance; these include mutants lacking the OM cytochromes OmcA and OmcB and the OM protein MtrB. Even though strains lacking OmcB and MtrB cannot reduce Fe(III) or AQDS, they still release a substance that restores the ability of CMA-1 to use MK-dependent electron acceptors, including AQDS and Fe(III). Quinone analysis showed that this released substance restores MK synthesis in CMA 1. This ability to restore MK synthesis in CMA-1 explains the cross-feeding results and challenges the previous hypothesis that this substance represents a redox shuttle that facilitates metal respiration. PMID- 15345429 TI - Simultaneous fluorescence in situ hybridization of mRNA and rRNA in environmental bacteria. AB - We developed for Bacteria in environmental samples a sensitive and reliable mRNA fluorescence in situ hybridization (FISH) protocol that allows for simultaneous cell identification by rRNA FISH. Samples were carbethoxylated with diethylpyrocarbonate to inactivate intracellular RNases and pretreated with lysozyme and/or proteinase K at different concentrations. Optimizing the permeabilization of each type of sample proved to be a critical step in avoiding false-negative or false-positive results. The quality of probes as well as a stringent hybridization temperature were determined with expression clones. To increase the sensitivity of mRNA FISH, long ribonucleotide probes were labeled at a high density with cis-platinum-linked digoxigenin (DIG). The hybrid was immunocytochemically detected with an anti-DIG antibody labeled with horseradish peroxidase (HRP). Subsequently, the hybridization signal was amplified by catalyzed reporter deposition with fluorochrome-labeled tyramides. p Iodophenylboronic acid and high concentrations of NaCl substantially enhanced the deposition of tyramides and thus increased the sensitivity of our approach. After inactivation of the antibody-delivered HRP, rRNA FISH was performed by following routine protocols. To show the broad applicability of our approach, mRNA of a key enzyme of aerobic methane oxidation, particulate methane monooxygenase (subunit A), was hybridized with different types of samples: pure cultures, symbionts of a hydrothermal vent bivalve, and even sediment, one of the most difficult sample types with which to perform successful FISH. By simultaneous mRNA FISH and rRNA FISH, single cells are identified and shown to express a particular gene. Our protocol is transferable to many different types of samples with the need for only minor modifications of fixation and permeabilization procedures. PMID- 15345430 TI - Clearance of human-pathogenic viruses from sludge: study of four stabilization processes by real-time reverse transcription-PCR and cell culture. AB - Sludges derived from wastewater treatment are foul-smelling, biologically unstable substances. As well as containing numerous pathogenic microorganisms, they also consist of organic matter that can be used as agricultural fertilizer. Legislation nevertheless requires sludges to be virologically tested prior to spreading by the counting of infectious enterovirus particles. This method, based on culture of enterovirus on BGM cells, is lengthy and not very sensitive. The aim of this study was to propose an alternative method of genome quantification for all enteroviruses that is applicable to verifying the elimination of viruses in complex samples such as sludges. Our complete protocol was compared to the official method, consisting of enterovirus enumeration with the most probable number of cythopathic unit (MPNCU) assay through the study of four stabilization procedures: liming, composting, heat treatment, and mesophile anaerobic digestion. Enterovirus quantities at the start of the stabilization procedures were between 37 and 288 MPNCU/g on the one scale and between 4 and 5 log genome copies/g on the other. It was shown that all procedures except mesophile anaerobic digestion were highly effective in the elimination of enterovirus particles and genomes in wastewater sludges. Reduction of viruses by mesophile anaerobic digestion was by only 1 log (infectious particles and genomes). In conclusion, stabilization processes can indeed be checked by virological quality control of sludges with gene amplification. However, the infectivity of genomes needs to be confirmed with cell culture or a correlation model if the virological risk inherent in the agricultural use of such sludges is to be fully addressed. PMID- 15345431 TI - Comparison of atomic force microscopy interaction forces between bacteria and silicon nitride substrata for three commonly used immobilization methods. AB - Atomic force microscopy (AFM) has emerged as a powerful technique for mapping the surface morphology of biological specimens, including bacterial cells. Besides creating topographic images, AFM enables us to probe both physicochemical and mechanical properties of bacterial cell surfaces on a nanometer scale. For AFM, bacterial cells need to be firmly anchored to a substratum surface in order to withstand the friction forces from the silicon nitride tip. Different strategies for the immobilization of bacteria have been described in the literature. This paper compares AFM interaction forces obtained between Klebsiella terrigena and silicon nitride for three commonly used immobilization methods, i.e., mechanical trapping of bacteria in membrane filters, physical adsorption of negatively charged bacteria to a positively charged surface, and glutaraldehyde fixation of bacteria to the tip of the microscope. We have shown that different sample preparation techniques give rise to dissimilar interaction forces. Indeed, the physical adsorption of bacterial cells on modified substrata may promote structural rearrangements in bacterial cell surface structures, while glutaraldehyde treatment was shown to induce physicochemical and mechanical changes on bacterial cell surface properties. In general, mechanical trapping of single bacterial cells in filters appears to be the most reliable method for immobilization. PMID- 15345432 TI - Identification of members of the metabolically active microbial populations associated with Beggiatoa species mat communities from Gulf of Mexico cold-seep sediments. AB - In this study, the composition of the metabolically active fraction of the microbial community occurring in Gulf of Mexico marine sediments (water depth, 550 to 575 m) with overlying filamentous bacterial mats was determined. The mats were mainly composed of either orange- or white-pigmented Beggiatoa spp. Complementary 16S ribosomal DNA (crDNA) was obtained from rRNA extracted from three different sediment depths (0 to 2, 6 to 8, and 10 to 12 cm) that had been subjected to reverse transcription-PCR amplification. Domain-specific 16S PCR primers were used to construct 12 different 16S crDNA libraries containing 333 Archaea and 329 Bacteria clones. Analysis of the Archaea clones indicated that all sediment depths associated with overlying orange- and white-pigmented microbial mats were almost exclusively dominated by ANME-2 (95% of total Archaea clones), a lineage related to the methanogenic order Methanosarcinales. In contrast, bacterial diversity was considerably higher, with the dominant phylotype varying by sediment depth. An equivalent number of clones detected at 0 to 2 cm, representing a total of 93%, were related to the gamma and delta classes of Proteobacteria, whereas clones related to delta-Proteobacteria dominated the metabolically active fraction of the bacterial community occurring at 6 to 8 cm (79%) and 10 to 12 cm (85%). This is the first phylogenetics-based evaluation of the presumptive metabolically active fraction of the Bacteria and Archaea community structure investigated along a sediment depth profile in the northern Gulf of Mexico, a hydrocarbon-rich cold-seep region. PMID- 15345433 TI - Geochemical rate-RNA integration study: ribulose-1,5-bisphosphate carboxylase/oxygenase gene transcription and photosynthetic capacity of planktonic photoautotrophs. AB - A pilot field experiment to assess the relationship between traditional biogeochemical rate measurements and transcriptional activity of microbial populations was carried out at the LEO 15 site off Tuckerton, N.J. Here, we report the relationship between photosynthetic capacity of autotrophic plankton and transcriptional activity of the large subunit gene (rbcL) for ribulose-1,5 bisphosphate carboxylase/oxygenase (RubisCO), the enzyme responsible for primary carbon fixation during photosynthesis. Similar diel patterns of carbon fixation and rbcL gene expression were observed in three of four time series, with maxima for photosynthetic capacity (P(max)) and rbcL mRNA occurring between 10 a.m. and 1 p.m. The lowest P(max) and rbcL levels were detected between 6 p.m. and 10:30 p.m. A significant correlation was found between P(max) and form ID rbcL mRNA (R(2) = 0.56) and forms IA and IB (R(2) = 0.41 and 0.47, respectively). The correlation between the abundance of "diatom" rbcL and P(max) mRNA was modest (R(2) = 0.49; n = 12) but improved dramatically (R(2) = 0.97; n = 10) upon removal of two outliers which represented afternoon samples with high P(max) but lower mRNA levels. Clone libraries from reverse transcription-PCR-amplified rbcL mRNA indicated the presence of several chromophytic algae (diatoms, prymnesiophytes, and chrysophytes) and some eukaryotic green flagellates. Analogous results were obtained from amplified small rRNA sequences and secondary pigment analysis. These results suggest that diatoms were a major contributor to carbon fixation at LEO 15 at the time of sampling and that photosynthetic carbon fixation was partially controlled by transcriptional regulation of the RubisCO gene. PMID- 15345434 TI - Effects of temperature and salinity on Vibrio vulnificus population dynamics as assessed by quantitative PCR. AB - The abundance of Vibrio vulnificus in coastal environments has been linked to water temperature, while its relationship to salinity is less clear. We have developed a culture-independent, most-probable-number quantitative PCR approach to examine V. vulnificus population dynamics in Barnegat Bay, N.J. Based on the combined analysis of our results from Barnegat Bay and from the literature, the present data show that (i) V. vulnificus population dynamics are strongly correlated to water temperature and (ii) although the general trend is for V. vulnificus abundance to be inversely correlated with salinity, this relationship depends on salinity levels. Irrespective of temperature, high abundances of V. vulnificus are observed at 5 to 10 ppt, which thus appears to be the optimal salinity regime for their survival. At 20 to 25 ppt, V. vulnificus abundances show a positive correlation to salinity. Unsuccessful attempts to resuscitate V. vulnificus, combined with our inability to detect cells during the winter despite an assay adapted to detect viable but nonculturable (VBNC) cells, suggest that the decline and eventual disappearance of V. vulnificus from the water column during the winter months is due primarily to a significant reduction in population size and is not only the consequence of cells entering the VBNC state. These findings are in line with the hypothesis that the sediment serves as a refuge for a subpopulation of V. vulnificus over the winter and weather-driven mixing events during the spring initiate a summer bloom in the water column. PMID- 15345435 TI - The pool of ADP and ATP regulates anaerobic product formation in resting cells of Lactococcus lactis. AB - Lactococcus lactis grows homofermentatively on glucose, while its growth on maltose under anaerobic conditions results in mixed acid product formation in which formate, acetate, and ethanol are formed in addition to lactate. Maltose was used as a carbon source to study mixed acid product formation as a function of the growth rate. In batch and nitrogen-limited chemostat cultures mixed acid product formation was shown to be linked to the growth rate, and homolactic fermentation occurred only in resting cells. Two of the four lactococcal strains investigated with maltose, L. lactis 65.1 and MG1363, showed more pronounced mixed acid product formation during growth than L. lactis ATCC 19435 or IL-1403. In resting cell experiments all four strains exhibited homolactic fermentation. In resting cells the intracellular concentrations of ADP, ATP, and fructose 1,6 bisphosphate were increased and the concentration of P(i) was decreased compared with the concentrations in growing cells. Addition of an ionophore (monensin or valinomycin) to resting cultures of L. lactis 65.1 induced mixed acid product formation concomitant with decreases in the ADP, ATP, and fructose 1,6 bisphosphate concentrations. ADP and ATP were shown to inhibit glyceraldehyde-3 phosphate dehydrogenase, lactate dehydrogenase, and alcohol dehydrogenase in vitro. Alcohol dehydrogenase was the most sensitive enzyme and was totally inhibited at an adenine nucleotide concentration of 16 mM, which is close to the sum of the intracellular concentrations of ADP and ATP of resting cells. This inhibition of alcohol dehydrogenase might be partially responsible for the homolactic behavior of resting cells. A hypothesis regarding the level of the ATP ADP pool as a regulating mechanism for the glycolytic flux and product formation in L. lactis is discussed. PMID- 15345436 TI - Integration of microbial ecology and statistics: a test to compare gene libraries. AB - Libraries of 16S rRNA genes provide insight into the membership of microbial communities. Statistical methods help to determine whether differences in library composition are artifacts of sampling or are due to underlying differences in the communities from which they are derived. To contribute to a growing statistical framework for comparing 16S rRNA libraries, we present a computer program, integral -LIBSHUFF, which calculates the integral form of the Cramer-von Mises statistic. This implementation builds upon the LIBSHUFF program, which uses an approximation of the statistic and makes a number of modifications that improve precision and accuracy. Once integral -LIBSHUFF calculates the P values, when pairwise comparisons are tested at the 0.05 level, the probability of falsely identifying a significant P value is 0.098 for a study with two libraries, 0.265 for three libraries, and 0.460 for four libraries. The potential negative effects of making the multiple pairwise comparisons necessitate correcting for the increased likelihood that differences between treatments are due to chance and do not reflect biological differences. Using integral -LIBSHUFF, we found that previously published 16S rRNA gene libraries constructed from Scottish and Wisconsin soils contained different bacterial lineages. We also analyzed the published libraries constructed for the zebrafish gut microflora and found statistically significant changes in the community during development of the host. These analyses illustrate the power of integral -LIBSHUFF to detect differences between communities, providing the basis for ecological inference about the association of soil productivity or host gene expression and microbial community composition. PMID- 15345437 TI - Regulation of the N-acyl homoserine lactone-dependent quorum-sensing system in rhizosphere Pseudomonas putida WCS358 and cross-talk with the stationary-phase RpoS sigma factor and the global regulator GacA. AB - Quorum sensing is a cell population-density dependent regulatory system which in gram-negative bacteria often involves the production and detection of N-acyl homoserine lactones (AHLs). Some Pseudomonas putida strains have been reported to produce AHLs, and one quorum-sensing locus has been identified. However, it appears that the majority of strains do not produce AHLs. In this study we report the identification and regulation of the AHL-dependent system of rhizosphere P. putida WCS358. This system is identical to the recently identified system of P. putida strain IsoF and very similar to the las system of Pseudomonas aeruginosa. It is composed of three genes, the luxI family member ppuI, the putative repressor rsaL, and the luxR family member ppuR. A genomic ppuR::Tn5 mutant of strain WCS358 was identified by its inability to produce AHLs when it was cross streaked in close proximity to an AHL biosensor, whereas an rsaL::Tn5 genomic mutant was identified by its ability to overproduce AHL molecules. Using transcriptional promoter fusions, we studied expression profiles of the rsaL, ppuI, and ppuR promoters in various genetic backgrounds. At the onset of the stationary phase, the autoinducer synthase ppuI gene expression is under positive regulation by PpuR-AHL and under negative regulation by RsaL, indicating that the molecules could be in competition for binding at the ppuI promoter. In genomic rsaL::Tn5 mutants ppuI expression and production of AHL levels increased dramatically; however, both processes were still under growth phase regulation, indicating that RsaL is not involved in repressing AHL production at low cell densities. The roles of the global response regulator GacA and the stationary phase sigma factor RpoS in the regulation of the AHL system at the onset of the stationary phase were also investigated. The P. putida WCS358 gacA gene was cloned and inactivated in the genome. It was determined that the three global regulatory systems are closely linked, with quorum sensing and RpoS regulating each other and GacA positively regulating ppuI expression. Studies of the regulation of AHL quorum-sensing systems have lagged behind other studies and are important for understanding how these systems are integrated into the overall growth phase and metabolic status of the cells. PMID- 15345438 TI - Cloning and characterization of gluconolactone oxidase of Penicillium cyaneo fulvum ATCC 10431 and evaluation of its use for production of D-erythorbic acid in recombinant Pichia pastoris. AB - A D-erythorbic acid-forming soluble flavoprotein, gluconolactone oxidase (GLO), was purified from Penicillium cyaneo-fulvum strain ATCC 10431 and partially sequenced. Peptide sequences were used to isolate a cDNA clone encoding the enzyme. The cloned gene exhibits high levels of similarity with the genes encoding other known eukaryotic lactone oxidases and also with the genes encoding some putative prokaryotic lactone oxidases. Analysis of the coding sequence of the GLO gene indicated the presence of a typical secretion signal sequence at the N terminus of GLO. No other targeting or anchoring signals were found, suggesting that GLO is the first known lactone oxidase that is secreted rather than targeted to the membranes of the endoplasmic reticulum or mitochondria. Experimental evidence, including the N-terminal sequence of mature GLO and data on glycosylation and localization of the enzyme in native and recombinant hosts, supports this analysis. The GLO gene was expressed in Pichia pastoris, and recombinant GLO was produced by using the strong methanol-induced AOX1 promoter. In order to evaluate the suitability of purified GLO for production of D erythorbic acid, we immobilized it on N-hydroxysuccinimide-activated Sepharose and found that the immobilized GLO retained full activity during immobilization but was rather unstable under reaction conditions. Our results show that both soluble and immobilized forms of GLO can, in principle, be used for production of D-erythorbic acid from D-glucono-delta-lactone or (in combination with glucose oxidase and catalase) from glucose. We also demonstrated the feasibility of glucose-D-erythorbic acid fermentation with recombinant strains coexpressing GLO and glucose oxidase genes, and we analyzed problems associated with construction of efficient D-erythorbic acid-producing hosts. PMID- 15345439 TI - Experimental and theoretical bases of specific affinity, a cytoarchitecture-based formulation of nutrient collection proposed to supercede the Michaelis-Menten paradigm of microbial kinetics. AB - A theory for solute uptake by whole cells was derived with a focus on the ability of oligobacteria to sequester nutrients. It provided a general relationship that was used to obtain the kinetic constants for in situ marine populations in the presence of naturally occurring substrates. In situ affinities of 0.9 to 400 liters g of cells(-1) h(-1) found were up to 10(3) times smaller than those from a "Marinobacter arcticus " isolate, but springtime values were greatly increased by warming. Affinities of the isolate for usual polar substrates but not for hydrocarbons were diminished by ionophores. A kinetic curve or Monod plot was constructed from the best available data for cytoarchitectural components of the isolate by using the theory together with concepts and calculations from first principles. The order of effect of these components on specific affinity was membrane potential > cytoplasmic enzyme concentration > cytoplasmic enzyme affinity > permease concentration > area of the permease site > translation coefficient > porin concentration. Component balance was influential as well; a small increase in cytoplasmic enzyme concentration gave a large increase in the effect of permease concentration. The effect of permease concentration on specific affinity was large, while the effect on K(m) was small. These results are in contrast to the Michaelis-Menten theory as applied by Monod that has uptake kinetics dependent on the quality of the permease molecules, with K(m) as an independent measure of affinity. Calculations demonstrated that most oligobacteria in the environment must use multiple substrates simultaneously to attain sufficient energy and material for growth, a requirement consistent with communities largely comprising few species. PMID- 15345440 TI - Phylogenetic analysis of polyketide synthase I domains from soil metagenomic libraries allows selection of promising clones. AB - The metagenomic approach provides direct access to diverse unexplored genomes, especially from uncultivated bacteria in a given environment. This diversity can conceal many new biosynthetic pathways. Type I polyketide synthases (PKSI) are modular enzymes involved in the biosynthesis of many natural products of industrial interest. Among the PKSI domains, the ketosynthase domain (KS) was used to screen a large soil metagenomic library containing more than 100,000 clones to detect those containing PKS genes. Over 60,000 clones were screened, and 139 clones containing KS domains were detected. A 700-bp fragment of the KS domain was sequenced for 40 of 139 randomly chosen clones. None of the 40 protein sequences were identical to those found in public databases, and nucleic sequences were not redundant. Phylogenetic analyses were performed on the protein sequences of three metagenomic clones to select the clones which one can predict to produce new compounds. Two PKS-positive clones do not belong to any of the 23 published PKSI included in the analysis, encouraging further analyses on these two clones identified by the selection process. PMID- 15345441 TI - Competition between ammonia-oxidizing bacteria and benthic microalgae. AB - The abundance, activity, and diversity of ammonia-oxidizing bacteria (AOB) were studied in prepared microcosms with and without microphytobenthic activity. In the microcosm without alga activity, both AOB abundance, estimated by real-time PCR, and potential nitrification increased during the course of the experiment. AOB present in the oxic zone of these sediments were able to fully exploit their nitrification potential because NH(4)(+) did not limit growth. In contrast, AOB in the alga-colonized sediments reached less than 20% of their potential activity, suggesting starvation of cells. Starvation resulted in a decrease with time in the abundance of AOB as well as in nitrification potential. This decrease was correlated with an increase in alga biomass, suggesting competitive exclusion of AOB by microalgae. Induction of N limitation in the oxic zone of the alga colonized sediments and O(2) limitation of the majority of AOB in darkness were major mechanisms by which microalgae suppressed the growth and survival of AOB. The competition pressure from the algae seemed to act on the entire population of AOB, as no differences were observed by denaturing gradient gel electrophoresis of amoA fragments during the course of the experiment. Enumeration of bacteria based on 16S rRNA gene copies and d-amino acids suggested that the algae also affected other bacterial groups negatively. Our data indicate that direct competitive interaction takes place between algae and AOB and that benthic algae are superior competitors because they have higher N uptake rates and grow faster than AOB. PMID- 15345442 TI - Characterization of a highly enriched dehalococcoides-containing culture that grows on vinyl chloride and trichloroethene. AB - A highly enriched culture that reductively dechlorinates trichloroethene (TCE), cis-1,2-dichloroethene (cDCE), and vinyl chloride (VC) to ethene without methanogenesis is described. The Dehalococcoides strain in this enrichment culture had a yield of (5.6 +/- 1.4) x 10(8) 16S rRNA gene copies/micromol of Cl( ) when grown on VC and hydrogen. Unlike the other VC-degrading cultures described in the literature, strains VS and BAV1, this culture maintained the ability to grow on TCE with a yield of (3.6 +/- 1.3) x 10(8) 16S rRNA gene copies/micromol of Cl(-). The yields on an electron-equivalent basis measured for the culture grown on TCE and on VC were not significantly different, indicating that both substrates supported growth equally well. PCR followed by denaturing gradient gel electrophoresis, cloning, and phylogenetic analyses revealed that this culture contained one Dehalococcoides 16S rRNA gene sequence, designated KB-1/VC, that was identical (over 1,386 bp) to the sequences of previously described organisms FL2 and CBDB1. A second Dehalococcoides sequence found in separate KB-1 enrichment cultures maintained on cDCE, TCE, and tetrachloroethene was no longer present in the VC-H(2) enrichment culture. This second Dehalococcoides sequence was identical to that of BAV1. As neither FL2 nor CBDB1 can dechlorinate VC to ethene in a growth-related fashion, it is clear that current 16S rRNA gene-based analyses do not provide sufficient information to distinguish between metabolically diverse members of the Dehalococcoides group. PMID- 15345443 TI - Lactococcal plasmid pNP40 encodes a novel, temperature-sensitive restriction modification system. AB - A novel restriction-modification system, designated LlaJI, was identified on pNP40, a naturally occurring 65-kb plasmid from Lactococcus lactis. The system comprises four adjacent similarly oriented genes that are predicted to encode two m(5)C methylases and two restriction endonucleases. The LlaJI system, when cloned into a low-copy-number vector, was shown to confer resistance against representatives of the three most common lactococcal phage species. This phage resistance phenotype was found to be strongly temperature dependent, being most effective at 19 degrees C. A functional analysis confirmed that the predicted methylase-encoding genes, llaJIM1 and llaJIM2, were both required to mediate complete methylation, while the assumed restriction enzymes, specified by llaJIR1 and llaJIR2, were both necessary for the complete restriction phenotype. A Northern blot analysis revealed that the four LlaJI genes are part of a 6-kb operon and that the relative abundance of the LlaJI-specific mRNA in the cells does not appear to contribute to the observed temperature-sensitive profile. This was substantiated by use of a LlaJI promoter-lacZ fusion, which further revealed that the LlaJI operon appears to be subject to transcriptional regulation by an as yet unidentified element(s) encoded by pNP40. PMID- 15345444 TI - Isolation and characterization of a rolling-circle-type plasmid from Rhodococcus erythropolis and application of the plasmid to multiple-recombinant-protein expression. AB - We isolated, sequenced, and characterized the cryptic plasmid pRE8424 from Rhodococcus erythropolis DSM8424. Plasmid pRE8424 is a 5,987-bp circular plasmid; it carries six open reading frames and also contains cis-acting elements, specifically a single-stranded origin and a double-stranded origin, which are characteristic of rolling-circle-replication plasmids. Experiments with pRE8424 derivatives carrying a mutated single-stranded origin sequence showed that single stranded DNA intermediates accumulated in the cells because of inefficient conversion from single-stranded DNA to double-stranded DNA. This result indicates that pRE8424 belongs to the pIJ101/pJV1 family of rolling-circle-replication plasmids. Expression vectors that are functional in several Rhodococcus species were constructed by use of the replication origin from pRE8424. We previously reported a cryptic plasmid, pRE2895, from R. erythropolis, which may replicate by a theta-type mechanism, like ColE2 plasmids. The new expression vectors originating from pRE8424 were compatible with those derived from pRE2895. Coexpression experiments with these compatible expression vectors indicated that the plasmids are suitable for the simultaneous expression of multiple recombinant proteins. PMID- 15345445 TI - Implicating the glutathione-gated potassium efflux system as a cause of electrophile-induced activated sludge deflocculation. AB - The glutathione-gated K(+) efflux (GGKE) system represents a protective microbial stress response that is activated by electrophilic or thiol-reactive stressors. It was hypothesized that efflux of cytoplasmic K(+) occurs in activated sludge communities in response to shock loads of industrially relevant electrophilic chemicals and results in significant deflocculation. Novosphingobium capsulatum, a bacterium consistent with others found in activated sludge treatment systems, responded to electrophilic thiol reactants with rapid efflux of up to 80% of its cytoplasmic K(+) pool. Furthermore, N. capsulatum and activated sludge cultures exhibited dynamic efflux-uptake-efflux responses very similar to those observed by others in Escherichia coli K-12 exposed to the electrophilic stressors N ethylmaleimide and 1-chloro-2,4-dinitrobenzene and the reducing agent dithiothreitol. Fluorescent LIVE/DEAD stains were used to show that cell lysis was not the cause of electrophile-induced K(+) efflux. Nigericin was used to artificially stimulate K(+) efflux from N. capsulatum and activated sludge cultures as a comparison to electrophile-induced K(+) efflux and showed that cytoplasmic K(+) efflux by both means corresponded with activated sludge deflocculation. These results parallel those of previous studies with pure cultures in which GGKE was shown to cause cytoplasmic K(+) efflux and implicate the GGKE system as a probable causal mechanism for electrophile-induced, activated sludge deflocculation. Calculations support the notion that shock loads of electrophilic chemicals result in very high K(+) concentrations within the activated sludge floc structure, and these K(+) levels are comparable to that which caused deflocculation by external (nonphysiological) KCl addition. PMID- 15345446 TI - Genetic structure of a Spodoptera frugiperda nucleopolyhedrovirus population: high prevalence of deletion genotypes. AB - A Nicaraguan field isolate (SfNIC) of Spodoptera frugiperda nucleopolyhedrovirus was purified by plaque assay on Sf9 cells. Nine distinct genotypes, A to I, were identified by their restriction endonuclease profiles. Variant SfNIC-B was selected as the standard because its restriction profile corresponded to that of the wild-type isolate. Physical maps were generated for each of the variants. The differences between variants and the SfNIC-B standard were confined to the region between map units 9 and 32.5. This region included PstI-G, PstI-F, PstI-L, PstI-K and EcoRI-L fragments. Eight genotypes presented a deletion in their genome compared with SfNIC-B. Occlusion body-derived virions of SfNIC-C, -D and -G accounted for 41% of plaque-purified clones. These variants were not infectious per os but retained infectivity by injection into S. frugiperda larvae. Median 50% lethal concentration values for the other cloned genotypes were significantly higher than that of the wild type. The variants also differed in their speed of kill. Noninfectious variants SfNIC-C and -D lacked the pif and pif-2 genes. Infectivity was restored to these variants by plasmid rescue with a plasmid comprising both pif and pif-2. Transcription of an SfNIC-G gene was detected by reverse transcription-PCR in insects, but no fatal disease developed. Transcription was not detected in SfNIC-C or -D-inoculated larvae. We conclude that the SfNIC population presents high levels of genetic diversity, localized to a 17-kb region containing pif and pif-2, and that interactions among complete and deleted genotypic variants will likely influence the capacity of this virus to control insect pests. PMID- 15345447 TI - Blakeslea trispora genes for carotene biosynthesis. AB - We cloned the carB and carRA genes involved in beta-carotene biosynthesis from overproducing and wild-type strains of Blakeslea trispora. The carB gene has a length of 1,955 bp, including two introns of 141 and 68 bp, and encodes a protein of 66.4 kDa with phytoene dehydrogenase activity. The carRA gene contains 1,894 bp, with a single intron of 70 bp, and encodes a protein of 69.6 kDa with separate domains for lycopene cyclase and phytoene synthase. The estimated transcript sizes for carB and carRA were 1.8 and 1.9 kb, respectively. CarB from the beta-carotene-overproducing strain B. trispora F-744 had an S528R mutation and a TAG instead of a TAA stop codon. The overproducing strain also had a P143S mutation in CarRA. Both B. trispora genes could complement mutations in orthologous genes in Mucor circinelloides and could be used to construct transformed strains of M. circinelloides that produced higher levels of beta carotene than did the nontransformed parent. The results show that these genes are conserved across the zygomycetes and that the B. trispora carB and carRA genes are functional and potentially useable to increase carotenoid production. PMID- 15345448 TI - Alkaline anaerobic respiration: isolation and characterization of a novel alkaliphilic and metal-reducing bacterium. AB - Iron-reducing enrichments were obtained from leachate ponds at the U.S. Borax Company in Boron, Calif. Based on partial small-subunit (SSU) rRNA gene sequences (approximately 500 nucleotides), six isolates shared 98.9% nucleotide identity. As a representative, the isolate QYMF was selected for further analysis. QYMF could be grown with Fe(III)-citrate, Fe(III)-EDTA, Co(III)-EDTA, or Cr(VI) as electron acceptors, and yeast extract and lactate could serve as electron donors. Growth during iron reduction occurred over the pH range of 7.5 to 11.0 (optimum, pH 9.5), a sodium chloride range of 0 to 80 g/liter (optimum, 20 g/liter), and a temperature range of 4 to 45 degrees C (optimum, approximately 35 degrees C), and iron precipitates were formed. QYMF was a strict anaerobe that could be grown in the presence of borax, and the cells were straight rods that produced endospores. Sodium chloride and yeast extract stimulated growth. Phylogenetic analysis of the SSU rRNA gene indicated that the bacterium was a low-G+C gram-positive microorganism and had 96 and 92% nucleotide identity with Alkaliphilus transvaalensis and Alkaliphilus crotonatoxidans, respectively. The major phospholipid fatty acids were 14:1, 16:1omega7c, and 16:0, which were different from those of other alkaliphiles but similar to those of reported iron-reducing bacteria. The results demonstrated that the isolate might represent a novel metal reducing alkaliphilic species. The name Alkaliphilus metalliredigens sp. nov. is proposed. The isolation and activity of metal-reducing bacteria from borax contaminated leachate ponds suggest that bioremediation of metal-contaminated alkaline environments may be feasible and have implications for alkaline anaerobic respiration. PMID- 15345449 TI - Cloning and characterization of the bile salt hydrolase genes (bsh) from Bifidobacterium bifidum strains. AB - Biochemical characterization of the purified bile salt hydrolase (BSH) from Bifidobacterium bifidum ATCC 11863 revealed some distinct characteristics not observed in other species of Bifidobacterium. The bsh gene was cloned from B. bifidum, and the DNA flanking the bsh gene was sequenced. Comparison of the deduced amino acid sequence of the cloned gene with previously known sequences revealed high homology with BSH enzymes from several microorganisms and penicillin V amidase (PVA) of Bacillus sphaericus. The proposed active sites of PVA were highly conserved, including that of the Cys-1 residue. The importance of the SH group in the N-terminal cysteine was confirmed by substitution of Cys with chemically and structurally similar residues, Ser or Thr, both of which resulted in an inactive enzyme. The transcriptional start point of the bsh gene has been determined by primer extension analysis. Unlike Bifidobacterium longum bsh, B. bifidum bsh was transcribed as a monocistronic unit, which was confirmed by Northern blot analysis. PCR amplification with the type-specific primer set revealed the high level of sequence homology in their bsh genes within the species of B. bifidum. PMID- 15345450 TI - Implications of amino acid substitutions in GyrA at position 83 in terms of oxolinic acid resistance in field isolates of Burkholderia glumae, a causal agent of bacterial seedling rot and grain rot of rice. AB - Oxolinic acid (OA), a quinolone, inhibits the activity of DNA gyrase composed of GyrA and GyrB and shows antibacterial activity against Burkholderia glumae. Since B. glumae causes bacterial seedling rot and grain rot of rice, both of which are devastating diseases, the emergence of OA-resistant bacteria has important implications on rice cultivation in Japan. Based on the MIC of OA, 35 B. glumae field isolates isolated from rice seedlings grown from OA-treated seeds in Japan were divided into sensitive isolates (OSs; 0.5 microg/ml), moderately resistant isolates (MRs; 50 microg/ml), and highly resistant isolates (HRs; > or =100 microg/ml). Recombination with gyrA of an OS, Pg-10, led MRs and HRs to become OA susceptible, suggesting that gyrA mutations are involved in the OA resistance of field isolates. The amino acid at position 83 in the GyrA of all OSs was Ser, but in all MRs and HRs it was Arg and Ile, respectively. Ser83Arg and Ser83Ile substitutions in the GyrA of an OS, Pg-10, resulted in moderate and high OA resistance, respectively. Moreover, Arg83Ser and Ile83Ser substitutions in the GyrA of MRs and HRs, respectively, resulted in susceptibility to OA. These results suggest that Ser83Arg and Ser83Ile substitutions in GyrA are commonly responsible for resistance to OA in B. glumae field isolates. PMID- 15345451 TI - Identification and functional characterization of a Xenorhabdus nematophila oligopeptide permease. AB - The bacterium Xenorhabdus nematophila is a mutualist of Steinernema carpocapsae nematodes and a pathogen of insects. Presently, it is not known what nutrients the bacterium uses to thrive in these host environments. In other symbiotic bacteria, oligopeptide permeases have been shown to be important in host interactions, and we therefore sought to determine if oligopeptide uptake is essential for growth or symbiotic functions of X. nematophila in laboratory or host environments. We identified an X. nematophila oligopeptide permease (opp) operon of two sequential oppA genes, predicted to encode oligopeptide-binding proteins, and putative permease-encoding genes oppB, oppC, oppD, and oppF. Peptide-feeding studies indicated that this opp operon encodes a functional oligopeptide permease. We constructed strains with mutations in oppA(1), oppA(2), or oppB and examined the ability of each mutant strain to grow in a peptide-rich laboratory medium and to interact with the two hosts. We found that the opp mutant strains had altered growth phenotypes in the laboratory medium and in hemolymph isolated from larval insects. However, the opp mutant strains were capable of initiating and maintaining both mutualistic and pathogenic host interactions. These data demonstrate that the opp genes allow X. nematophila to utilize peptides as a nutrient source but that this function is not essential for the existence of X. nematophila in either of its host niches. To our knowledge, this study represents the first experimental analysis of the role of oligopeptide transport in mediating a mutualistic invertebrate-bacterium interaction. PMID- 15345452 TI - Molecular fingerprinting of dairy microbial ecosystems by use of temporal temperature and denaturing gradient gel electrophoresis. AB - Numerous microorganisms, including bacteria, yeasts, and molds, constitute the complex ecosystem present in milk and fermented dairy products. Our aim was to describe the bacterial ecosystem of various cheeses that differ by production technology and therefore by their bacterial content. For this purpose, we developed a rapid, semisystematic approach based on genetic profiling by temporal temperature gradient electrophoresis (TTGE) for bacteria with low-G+C-content genomes and denaturing gradient gel electrophoresis (DGGE) for those with medium- and high-G+C-content genomes. Bacteria in the unknown ecosystems were assigned an identity by comparison with a comprehensive bacterial reference database of approximately 150 species that included useful dairy microorganisms (lactic acid bacteria), spoilage bacteria (e.g., Pseudomonas and Enterobacteriaceae), and pathogenic bacteria (e.g., Listeria monocytogenes and Staphylococcus aureus). Our analyses provide a high resolution of bacteria comprising the ecosystems of different commercial cheeses and identify species that could not be discerned by conventional methods; at least two species, belonging to the Halomonas and Pseudoalteromonas genera, are identified for the first time in a dairy ecosystem. Our analyses also reveal a surprising difference in ecosystems of the cheese surface versus those of the interior; the aerobic surface bacteria are generally G+C rich and represent diverse species, while the cheese interior comprises fewer species that are generally low in G+C content. TTGE and DGGE have proven here to be powerful methods to rapidly identify a broad range of bacterial species within dairy products. PMID- 15345453 TI - Raw cow milk bacterial population shifts attributable to refrigeration. AB - We monitored the dynamic changes in the bacterial population in milk associated with refrigeration. Direct analyses of DNA by using temporal temperature gel electrophoresis (TTGE) and denaturing gradient gel electrophoresis (DGGE) allowed us to make accurate species assignments for bacteria with low-GC-content (low GC%) (<55%) and medium- or high-GC% (>55%) genomes, respectively. We examined raw milk samples before and after 24-h conservation at 4 degrees C. Bacterial identification was facilitated by comparison with an extensive bacterial reference database ( approximately 150 species) that we established with DNA fragments of pure bacterial strains. Cloning and sequencing of fragments missing from the database were used to achieve complete species identification. Considerable evolution of bacterial populations occurred during conservation at 4 degrees C. TTGE and DGGE are shown to be a powerful tool for identifying the main bacterial species of the raw milk samples and for monitoring changes in bacterial populations during conservation at 4 degrees C. The emergence of psychrotrophic bacteria such as Listeria spp. or Aeromonas hydrophila is demonstrated. PMID- 15345454 TI - Metabolic primers for detection of (Per)chlorate-reducing bacteria in the environment and phylogenetic analysis of cld gene sequences. AB - Natural attenuation of the environmental contaminant perchlorate is a cost effective alternative to current removal methods. The success of natural perchlorate remediation is dependent on the presence and activity of dissimilatory (per)chlorate-reducing bacteria (DPRB) within a target site. To detect DPRB in the environment, two degenerate primer sets targeting the chlorite dismutase (cld) gene were developed and optimized. A nested PCR approach was used in conjunction with these primer sets to increase the sensitivity of the molecular detection method. Screening of environmental samples indicated that all products amplified by this method were cld gene sequences. These sequences were obtained from pristine sites as well as contaminated sites from which DPRB were isolated. More than one cld phylotype was also identified from some samples, indicating the presence of more than one DPRB strain at those sites. The use of these primer sets represents a direct and sensitive molecular method for the qualitative detection of (per)chlorate-reducing bacteria in the environment, thus offering another tool for monitoring natural attenuation. Sequences of cld genes isolated in the course of this project were also generated from various DPRB and provided the first opportunity for a phylogenetic treatment of this metabolic gene. Comparisons of the cld and 16S ribosomal DNA (rDNA) gene trees indicated that the cld gene does not track 16S rDNA phylogeny, further implicating the possible role of horizontal transfer in the evolution of (per)chlorate respiration. PMID- 15345455 TI - Microbial species involved in production of 1,2-sn-diacylglycerol and effects of phosphatidylcholine on human fecal microbiota. AB - 1,2-sn-Diacylglycerols (DAGs) are activators of protein kinase C (PKC), which is involved in the regulation of colonic mucosal proliferation. Extracellular DAG has been shown to stimulate the growth of cancer cell lines in vitro and may therefore play an important role in tumor promotion. DAG has been detected in human fecal extracts and is thought to be of microbial origin. Hitherto, no attempts have been made to identify the predominant fecal bacterial species involved in its production. We therefore used anaerobic batch culture systems to determine whether fecal bacteria could utilize phosphatidylcholine (0.5% [wt/vol]) to produce DAG. Production was found to be dependent upon the presence of the substrate and was enhanced in the presence of high concentrations of deoxycholate (5 and 10 mM) in the growth medium. Moreover, its production increased with the pH, and large inter- and intraindividual variations were observed between cultures seeded with inocula from different individuals. Clostridia and Escherichia coli multiplied in the fermentation systems, indicating their involvement in phosphatidylcholine metabolism. On the other hand, there was a significant decrease in the number of Bifidobacterium spp. in the presence of phosphatidylcholine. Pure-culture experiments showed that 10 of the 12 strains yielding the highest DAG levels (>50 nmol/ml) were isolated from batch culture enrichments run at pH 8.5. We found that the strains capable of producing large amounts of DAG were predominantly Clostridium bifermentans (8 of 12), followed by Escherichia coli (2 of 12). Interestingly, one DAG-producing strain was Bifidobacterium infantis, which is often considered a beneficial gut microorganism. Our results have provided further evidence that fecal bacteria can produce DAG and that specific bacterial groups are involved in this process. Future strategies to reduce DAG formation in the gut should target these species. PMID- 15345456 TI - Electrophoretic Mobility of Mycobacterium avium Complex Organisms. AB - The electrophoretic mobilities (EPMs) of 30 Mycobacterium avium complex organisms were measured. The EPMs of 15 clinical isolates ranged from -1.9 to -5.0 microM cm V(-1) s(-1), and the EPMs of 15 environmental isolates ranged from -1.9 to 4.6 microM cm V(-1) s(-1) at pH 7. PMID- 15345457 TI - Inhibition of Listeria monocytogenes in fish and meat systems by use of oregano and cranberry phytochemical synergies. AB - Optimized phenolics from oregano and cranberry extracts were evaluated for antimicrobial activity against Listeria monocytogenes in laboratory media and in beef and fish. The antimicrobial activity increased when oregano and cranberry extracts were mixed at a ratio of 75% oregano and 25% cranberry (wt/wt) with 0.1 mg of phenolic per disk or ml, and the efficacy was further enhanced by lactic acid. The inhibition by phytochemical and lactic acid synergies was most effective when beef and fish slices were stored at 4 degrees C. PMID- 15345458 TI - Potential role for fish in transmission of Mycobacterium ulcerans disease (Buruli ulcer): an environmental study. AB - This study reports a potential role that fish may play in the transmission of Mycobacterium ulcerans disease (Buruli ulcer). Fish found positive for M. ulcerans DNA all appear to feed on insects or plankton and are believed to concentrate M. ulcerans from this usual food source. These observations provide additional data supporting our previous hypothesis on sources of M. ulcerans and modes of transmission. PMID- 15345459 TI - Effect of Escherichia coli morphogene bolA on biofilms. AB - Biofilm physiology is established under a low growth rate. The morphogene bolA is mostly expressed under stress conditions or in stationary phase, suggesting that bolA could be implicated in biofilm development. In order to verify this hypothesis, we tested the effect of bolA on biofilm formation. Overexpression of bolA induces biofilm development, while bolA deletion decreases biofilms. PMID- 15345460 TI - Increasing the carbon flux toward synthesis of short-chain-length--medium-chain length polyhydroxyalkanoate in the peroxisome of Saccharomyces cerevisiae through modification of the beta-oxidation cycle. AB - Short-chain-length-medium-chain-length polyhydroxyalkanoates were synthesized in Saccharomyces cerevisiae from intermediates of the beta-oxidation cycle by expressing the polyhydroxyalkanoate synthases from Aeromonas caviae and Ralstonia eutropha in the peroxisomes. The quantity of polymer produced was increased by using a mutant of the beta-oxidation-associated multifunctional enzyme with low dehydrogenase activity toward R-3-hydroxybutyryl coenzyme A. PMID- 15345461 TI - Methanogenic inhibition by arsenic compounds. AB - The acute acetoclastic methanogenic inhibition of several inorganic and organic arsenicals was assayed. Trivalent species, i.e., methylarsonous acid and arsenite, were highly inhibitory, with 50% inhibitory concentrations of 9.1 and 15.0 microM, respectively, whereas pentavalent species were generally nontoxic. The nitrophenylarsonate derivate, roxarsone, displayed moderate toxicity. PMID- 15345462 TI - Fluorogenic selective and differential medium for isolation of Enterobacter sakazakii. AB - 4-Methylumbelliferyl-alpha-D-glucoside, the fluorogenic substrate of alpha glucosidase, was used as a selective marker to develop a differential medium for Enterobacter sakazakii. This bacterium showed strong fluorogenic characteristics clearly distinguishable from other microorganisms. On the basis of reducing background noise, an optimum basal medium and nitrogen source were selected. Incubation conditions were optimized. PMID- 15345463 TI - Rapid estimation of numbers of fecal Bacteroidetes by use of a quantitative PCR assay for 16S rRNA genes. AB - Assessment of health risk associated with fecal pollution requires a reliable fecal indicator and a rapid quantification method. We report the development of a Taq nuclease assay for enumeration of 16S rRNA genes of Bacteroidetes. Sensitivity and correlation with standard fecal indicators provide experimental evidence for application of the assay in monitoring fecal pollution. PMID- 15345464 TI - Large-scale population structure of human commensal Escherichia coli isolates. AB - The study of several Escherichia coli intestinal commensal isolates per individual in 265 healthy human subjects belonging to seven populations distributed worldwide showed that the E. coli population is highly structured, with major differences between the tropical and temperate populations. PMID- 15345465 TI - PCR-based identification of hyperthermophilic archaea of the family Thermococcaceae. AB - A method for rapid detection and identification of hyperthermophilic archaea of the family Thermococcaceae based on PCR amplification of 16S rRNA gene fragments with primers TcPc 173F (5'-TCCCCCATAGGYCTGRGGTACTGGAAGGTC-3') and TcPc 589R (5' GCCGTGRGATTTCGCCAGGGACTTACGGGC-3') was developed and used for identification of new isolates. PMID- 15345466 TI - Production of minicellulosomes from Clostridium cellulovorans in Bacillus subtilis WB800. AB - Two genes encoding EngB endoglucanase and mini-CbpA1 scaffolding protein of Clostridium cellulovorans were constructed and coexpressed in Bacillus subtilis WB800. The resulting minicellulosomes were isolated by gel filtration chromatography and characterized. Biochemical and immunological evidence indicated that fraction II contained minicellulosomes consisting of mini-CbpA1 and EngB. The in vivo synthesis of minicellulosomes suggests that it will be possible in the future to insert into B. subtilis cellulosomal genes that will allow growth on cellulosic materials and the production of various designer cellulosomes with specific functions. PMID- 15345467 TI - Widespread occurrence of a novel division of bacteria identified by 16S rRNA gene sequences originally found in deep marine sediments. AB - Phylogenetic analysis of 16S rRNA gene sequences from deep marine sediments identified a deeply branching clade, designated candidate division JS1. Primers for PCR amplification of partial 16S rRNA genes that target the JS1 division were developed and used to detect JS1 sequences in DNA extracted from various sedimentary environments, including, for the first time, coastal marine and brackish sediments. PMID- 15345468 TI - Inhibition of ACh-stimulated exocytosis by NSAIDs in guinea pig antral mucous cells: autocrine regulation of mucin secretion by PGE2. AB - The effects of indomethacin (IDM) and aspirin (ASA) on ACh (10 microM) stimulated exocytotic events were studied in guinea pig antral mucous cells by using video optical microscopy. IDM or ASA, which inhibits cyclooxygenase (COX), decreased the frequency of ACh-stimulated exocytotic events by 30% or 60%, respectively. The extent of inhibition induced by ASA (60%) decreased by 30% when IDM or arachidonic acid (AA, the substrate of COX) was added. IDM, unlike ASA, appears to induce the accumulation of AA, which enhances the frequency of ACh stimulated exocytotic events in ASA-treated cells. ONO-8713 (100 microM; an inhibitor of the EP1-EP4 prostaglandin receptors) and N-[2-((p bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide, HCl (H-89, 20 microM; an inhibitor of PKA) also decreased the frequency of ACh-stimulated exocytotic events by 60%. However, the supplementation of PGE(2) (1 microM) prevented the IDM-induced decrease in the frequency of ACh-stimulated exocytotic events. SC-560 (an inhibitor of COX-1) decreased the frequency of ACh-stimulated exocytotic events by 30%, but NS-398 (an inhibitor of COX-2) did not. Moreover, IDM decreased the frequency of exocytotic events stimulated by ionomycin, suggesting that COX-1 activity is stimulated by an increase in intracellular Ca(2+) concentration ([Ca(2+)](i)). ACh and ionomycin increased PGE(2) release in antral mucosal cells. In conclusion, in ACh-stimulated antral mucous cells, an increase in [Ca(2+)](i) activates Ca(2+)-regulated exocytotic events and PGE(2) release mediated by COX-1. The released PGE(2) induces the accumulation of cAMP, which enhances the Ca(2+)-regulated exocytosis. The autocrine mechanism mediated by PGE(2) maintains the high-level mucin release from antral mucous cells during ACh stimulation. PMID- 15345469 TI - Activation of proteinase-activated receptor-1 inhibits neurally evoked chloride secretion in the mouse colon in vitro. AB - The proteinase-activated thrombin receptor-1 (PAR-1) belongs to a unique family of G protein-coupled receptors activated by proteolytic cleavage. We studied the effect of PAR-1 activation in the regulation of ion transport in mouse colon in vitro. Expression of PAR-1 in mouse colon was assessed by RT-PCR and immunohistochemistry. To study the role of PAR-1 activation in chloride secretion, mouse colon was mounted in Ussing chambers. Changes in short-circuit current (Isc) were measured in tissues exposed to either thrombin, saline, the PAR-1-activating peptide TFLLR-NH2, or the inactive reverse peptide RLLFT-NH2, before electrical field stimulation (EFS). Experiments were repeated in the presence of either a PAR-1 antagonist or in PAR-1-deficient mice to assess receptor specificity. In addition, studies were conducted in the presence of chloride-free buffer or the muscarinic antagonist atropine to assess chloride dependency and the role of cholinergic neurons in the PAR-1-induced effect. PAR-1 mRNA was expressed in full-thickness specimens and mucosal scrapings of mouse colon. PAR-1 immunoreactivity was found on epithelial cells and on neurons in submucosal ganglia where it was colocalized with both VIP and neuropeptide Y. After PAR-1 activation by thrombin or TFLLR-NH2, secretory responses to EFS but not those to forskolin or carbachol were significantly reduced. The reduction in the response to EFS was not observed in the presence of the PAR-1 antagonist, in PAR-1-deficient mice, when chloride was excluded from the bathing medium, or when atropine was present. PAR-1 is expressed in submucosal ganglia in the mouse colon and its activation leads to a decrease in neurally evoked epithelial chloride secretion. PMID- 15345470 TI - Reduced hepatic extraction of palmitate in steatosis correlated to lower level of liver fatty acid binding protein. AB - Nonalcoholic fatty liver disease is the most common of all liver diseases. The hepatic disposition [(3)H]palmitate and its low-molecular-weight metabolites in perfused normal and steatotic rat liver were studied using the multiple indicator dilution technique and a physiologically based slow diffusion/bound pharmacokinetic model. The steatotic rat model was established by administration of 17 alpha-ethynylestradiol to female Wistar rats. Serum biochemistry markers and histology of treated and normal animals were assessed and indicated the presence of steatosis in the treatment group. The steatotic group showed a significantly higher alanine aminotransferase-to-aspartate aminotransferase ratio, lower levels of liver fatty acid binding protein and cytochrome P-450, as well as microvesicular steatosis with an enlargement of sinusoidal space. Hepatic extraction for unchanged [(3)H]palmitate and production of low-molecular-weight metabolites were found to be significantly decreased in steatotic animals. Pharmacokinetic analysis suggested that the reduced extraction and sequestration for palmitate and its metabolites was mainly attributed to a reduction in liver fatty acid binding protein in steatosis. PMID- 15345471 TI - Decreased epoxygenase and increased epoxide hydrolase expression in the mesenteric artery of obese Zucker rats. AB - Previous studies suggest that epoxyeicosatrienoic acids (EETs) are vasodilators of the mesenteric artery; however, the production and regulation of EETs in the mesenteric artery remain unclear. The present study was designed 1) to determine which epoxygenase isoform may contribute to formation of EETs in mesenteric arteries and 2) to determine the regulation of mesenteric artery cytochrome P-450 (CYP) enzymes in obese Zucker rats. Microvessels were incubated with arachidonic acid, and CYP enzyme activity was determined. Mesenteric arteries demonstrate detectable epoxygenase and hydroxylase activities. Next, protein and mRNA expressions were determined in microvessels. Although renal microvessels express CYP2C23 mRNA and protein, mesenteric arteries lacked CYP2C23 expression. CYP2C11 and CYP2J mRNA and protein were expressed in mesenteric arteries and renal microvessels. In addition, mesenteric artery protein expression was evaluated in lean and obese Zucker rats. Compared with lean Zucker rats, mesenteric arterial CYP2C11 and CYP2J proteins were decreased by 38 and 43%, respectively, in obese Zucker rats. In contrast, soluble epoxide hydrolase mRNA and protein expressions were significantly increased in obese Zucker rat mesenteric arteries. In addition, nitric oxide-independent dilation evoked by acetylcholine was significantly attenuated in mesenteric arteries of obese Zucker rats. These data suggest that the main epoxygenase isoforms expressed in mesenteric arteries are different from those expressed in renal microvessels and that decreased epoxygenases and increased soluble epoxide hydrolase are associated with impaired mesenteric artery dilator function in obese Zucker rats. PMID- 15345472 TI - Temporal expression profiles of organic anion transport proteins in placenta and fetal liver of the rat. AB - Physiological cholestasis linked to immature hepatobiliary transport systems for organic anions occurs in rat and human neonates. In utero, the placenta facilitates vectorial transfer of certain fetal-derived solutes to the maternal circulation for elimination. We compared the ontogenesis of organic anion transporters in the placenta and the fetal liver of the rat to assess their relative abundance throughout gestation and to determine whether the placenta compensates for the late maturation of transporters in the developing liver. The mRNA of members of the organic anion transporting polypeptide (Oatp) superfamily, the multidrug resistance protein (Mrp) family, one organic anion transporter (OAT), and the bile acid carriers Na(+)-taurocholate cotransporting polypeptide (Ntcp) and bile salt export pump (Bsep) was quantified by real-time PCR. The most abundant placental transporters were Oatp4a1, whose mRNA increased 10-fold during gestation, and Mrp1. Mrp1 immunolocalized predominantly to epithelial cells of the endoplacental yolk sac, suggesting an excretory role that sequesters fetal derived solutes in the yolk sac cavity, and faintly to the basal syncytiotrophoblast surface. The mRNA levels of Oatp2b1, Mrp3, and Bsep in the placenta exceeded those in the fetal liver until day 20 of gestation, suggesting that the fetus relies on placental clearance of substrates when expression in the developing liver is low. Mrp3 immunolocalized to the epithelium of the endoplacental yolk sac and less abundantly in the labyrinth zone and endothelium of the maternal arteries. The placental expression of Oatp1a1, Oatp1a4, Oatp1a5, Oatp1b2, Oat, Ntcp, Mrp2, and Mrp6 was low. PMID- 15345473 TI - Vertebrate phylogeny of hydrogen sulfide vasoactivity. AB - Hydrogen sulfide (H(2)S) is a recently identified endogenous vasodilator in mammals. In steelhead/rainbow trout (Oncorhynchus mykiss, Osteichthyes), H(2)S produces both dose-dependent dilation and a unique dose-dependent constriction. In this study, we examined H(2)S vasoactivity in all vertebrate classes to determine whether H(2)S is universally vasoactive and to identify phylogenetic and/or environmental trends. H(2)S was generated from NaHS and examined in unstimulated and precontracted systemic and, when applicable, pulmonary arteries (PA) from Pacific hagfish (Eptatretus stouti, Agnatha), sea lamprey (Petromyzon marinus, Agnatha), sandbar shark (Carcharhinus milberti, Chondrichthyes), marine toad (Bufo marinus, Amphibia), American alligator (Alligator mississippiensis, Reptilia), Pekin duck (Anas platyrhynchos domesticus, Aves), and white rat (Rattus rattus, Mammalia). In otherwise unstimulated vessels, NaHS produced 1) a dose-dependent relaxation in Pacific hagfish dorsal aorta; 2) a dose-dependent contraction in sea lamprey dorsal aorta, marine toad aorta, alligator aorta and PA, duck aorta, and rat thoracic aorta; 3) a threshold relaxation in shark ventral aorta, dorsal aorta, and afferent branchial artery; and 4) a multiphasic contraction-relaxation-contraction in the marine toad PA, duck PA, and rat PA. Precontraction of these vessels with another agonist did not affect the general pattern of NaHS vasoactivity with the exception of the rat aorta, where relaxation was now dominant. These results show that H(2)S is a phylogenetically ancient and versatile vasoregulatory molecule that appears to have been opportunistically engaged to suit both organ-specific and species-specific homeostatic requirements. PMID- 15345474 TI - Excitatory convergence of periaqueductal gray and somatic afferents in the solitary tract nucleus: role for neurokinin 1 receptors. AB - Our previous studies (Boscan P, Kasparov S, and Paton JF. Eur J Neurosci 16: 907 920, 2002) showed that activation of somatic afferents attenuated the baroreceptor reflex via neurokinin type 1 (NK(1)) and GABA(A) receptors within the nucleus of the solitary tract (NTS). The periaqueductal gray matter (PAG) can also depress baroreceptor reflex function and project to the NTS. In the present study, we have tested the possibility that the dorsolateral (dl)-PAG projects to the NTS neurons that also respond to somatic afferent input. In an in situ, arterially perfused, unanesthetized decerebrate rat preparation, somatic afferents (brachial plexus), cervical spinal cord, and dl-PAG were stimulated electrically, whereas NTS neurons were recorded extracellularly. From 45 NTS neurons excited by either brachial plexus or dl-PAG stimulation, 41 received convergence excitatory inputs from both afferents. Onset latency and evoked peak discharge frequency from brachial plexus afferents were 39.4 +/- 4.7 ms and 10.7 +/- 1.1 Hz, whereas this was 43.9 +/- 6.4 ms and 7.9 +/- 1 Hz, respectively, following dl-PAG stimulation. As revealed by using a paired pulse stimulation protocol, monosynaptic connections were found in 9 of 36 neurons tested from both spinal cord and dl-PAG. We tested NK(1)-receptor sensitivity in 38 neurons that received convergent inputs from brachial plexus/PAG. Fifteen neurons were sensitive to selective antagonism of NK(1) receptors. CP-99994, the NK(1) antagonist, failed to alter ongoing firing activity but reduced the evoked peak discharge frequency following stimulation of both brachial plexus (from 12.3 +/- 1.8 to 7.2 +/- 1.3 Hz; P < 0.01) and PAG (from 7.8 +/- 1.5 to 4.5 +/- 1 Hz; P < 0.01). We conclude that 1) somatic brachial and PAG afferents can converge onto single NTS neurons; 2) this convergence occurs via either direct or indirect pathways; and 3) NK(1) receptors are activated by some of these inputs. PMID- 15345475 TI - Actions of neuropeptide W in paraventricular hypothalamus: implications for the control of stress hormone secretion. AB - Neuropeptide W (NPW) is produced in neurons located in hypothalamus and brain stem, and its receptors are present in the hypothalamus, in particular in the paraventricular nucleus (PVN). Intracerebroventricular (ICV) administration of NPW activated, in a dose-related fashion, the hypothalamic-pituitary-adrenal axis, as determined by plasma corticosterone levels in conscious rats but, at those same doses, did not stimulate the release of oxytocin or vasopressin into the peripheral circulation or alter blood pressure or heart rate. The ability of ICV-administered NPW to stimulate the hypothalamic-pituitary-adrenal axis in conscious male rats was blocked by intravenous pretreatment with a corticotropin releasing hormone antagonist. This suggested an action of NPW in the parvocellular division of the PVN. Indeed, in hypothalamic slice preparations (whole cell patch recording), bath application of NPW depolarized and increased the spike frequency of the majority of electrophysiologically identified putative neuroendocrine PVN neurons. Effects on membrane potential were maintained in the presence of TTX, suggesting them to be direct postsynaptic actions on these neuroendocrine cells. Our data suggest that endogenous NPW, produced in brain, may play a physiologically relevant role in the neuroendocrine response to stress. PMID- 15345476 TI - Reduced plasma volume and mesenteric vascular reactivity in obese Zucker rats. AB - Obese Zucker rats (OZR) are mildly hypertensive with an apparently elevated sympathetic vasomotor tone compared with lean Zucker rats (LZR). Studies have also suggested enhanced adrenergic pressor reactivity in OZR but assumed comparable baroreflexes, or blood volume-to-body weight ratio, to LZR. In 15-wk old OZR and LZR, we measured plasma volume and vascular reactivity to norepinephrine (NE) and phenylephrine (PE) with doses evaluated by body weight and plasma volume. Plasma volume measured by dye dilution (Evans blue; 200 microl of 0.5%) showed that OZR had comparable blood volumes to LZR but lower blood volume-to-body weight ratio (3.4 +/- 0.2 ml/100 g) than LZR (5.7 +/- 0.2 ml/100 g, P < 0.05). Ganglionic blockade (mecamylamine, 4 mg/kg) in isoflurane anesthetized rats produced larger decreases in arterial pressure in OZR compared with LZR (52 +/- 2 vs. 46 +/- 2 mmHg). Pressor responses to NE (0.01-10 microg/kg) were exaggerated with doses analyzed by body weight but not analyzed by drug quantity. Pressor responses to PE (1-24 microg/kg) showed no difference with doses analyzed by body weight, but, analyzed by drug quantity, OZR showed a slight decrease in pressor reactivity. PE-induced increases in vascular resistance were exaggerated in the hindlimb circulation of OZR, normal in the renal circulation, and attenuated in the mesenteric circulation. The timing of the peak pressor response to PE corresponded with the increase in mesenteric vascular resistance, followed by rises in hindlimb and renal resistance. These data suggest that systemic adrenergic pressor reactivity is not enhanced in OZR, despite exaggerated vascular reactivity in the hindlimb of the OZR. PMID- 15345477 TI - Pharmacological preconditioning with resveratrol: role of CREB-dependent Bcl-2 signaling via adenosine A3 receptor activation. AB - Recent studies demonstrated that resveratrol, a grape-derived polyphenolic phytoalexin, provides pharmacological preconditioning (PC) of the heart through a NO-dependent mechanism. Because adenosine receptors play a role in PC, we examined whether they play any role in resveratrol PC. Rats were randomly assigned to groups perfused for 15 min with 1) Krebs-Henseleit bicarbonate buffer (KHB) only; 2) KHB containing 10 microM resveratrol; 3) 10 microM resveratrol + 1 microM 8-cyclopentyl-1,3-dimethylxanthine (CPT; adenosine A(1) receptor blocker); 4) 10 microM resveratrol + 1 microM 8-(3-chlorostyryl)caffeine (CSC; adenosine A(2a) receptor blocker); 5) 10 microM resveratrol + 1 microM 3-ethyl-5-benzyl-2 methyl-4-phenylethynyl-6-phenyl-1,4-(+/-)-dihydropyridine-3,5-dicarboxylate (MRS 1191; adenosine A(3) receptor blocker); or 6) 10 microM resveratrol + 3 microM 2 (4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride [LY-294002, phosphatidylinositol (PI)3-kinase inhibitor], and groups perfused with adenosine receptor blockers alone. Hearts were then subjected to 30-min ischemia followed by 2-h reperfusion. The results demonstrated significant cardioprotection with resveratrol evidenced by improved ventricular recovery and reduced infarct size and cardiomyocyte apoptosis. CPT and MRS 1191, but not CSC, abrogated the cardioprotective abilities of resveratrol, suggesting a role of adenosine A(1) and A(3) receptors in resveratrol PC. Resveratrol induced expression of Bcl-2 and caused its phosphorylation along with phosphorylation of cAMP response element binding protein (CREB), Akt, and Bad. CPT blocked phosphorylation of Akt and Bad without affecting CREB, whereas MRS 1191 blocked phosphorylation of all compounds, including CREB. LY-294002 partially blocked the cardioprotective abilities of resveratrol. The results indicate that resveratrol preconditions the heart through activation of adenosine A(1) and A(3) receptors, the former transmitting a survival signal through PI3-kinase-Akt-Bcl-2 signaling pathway and the latter protecting the heart through a CREB-dependent Bcl-2 pathway in addition to an Akt-Bcl-2 pathway. PMID- 15345478 TI - Profibrotic influence of high glucose concentration on cardiac fibroblast functions: effects of losartan and vitamin E. AB - Long-standing diabetes can result in the development of cardiomyopathy, which can be accompanied by myocardial fibrosis. Although exposure of cultured kidney and skin fibroblasts to high glucose (HG) concentration is known to increase collagen synthesis, little is known about cardiac fibroblasts (CFs). Therefore, we determined the influence of HG conditions on CF functions and the effects of losartan and vitamin E in these responses. We cultured rat CFs in either normal glucose (NG; 5.5 mM) or HG (25 mM) media and assessed changes in protein and collagen synthesis, matrix metalloproteinase (MMP) activity, and levels of mRNA for ANG II type 1 (AT(1)) receptors. Results indicate that HG-level CFs synthesized more protein and collagen, and these effects were not due to changes in osmotic pressure. The addition of ANG II stimulated protein and collagen synthesis in NG-concentration but not HG-concentration CFs. Interestingly, losartan pretreatment blocked the HG- or ANG II-induced increases in both protein and collagen synthesis. HG or ANG II decreased total MMP activity. Decreases in MMP activity were blocked by losartan. AT(1) mRNA levels were upregulated with HG concentration. Vitamin E pretreatment blocked the effects of HG on total protein synthesis and stimulated MMP activity. Results suggest that HG levels may promote fibrosis by increasing CF protein and collagen synthesis and decreasing MMP activity. HG levels may cause these effects via the upregulation of AT(1) receptors, which can be blocked by losartan. However, vitamin E can alter HG concentration-induced changes in CF functions independently of AT(1) mRNA levels. PMID- 15345479 TI - Direct observation of epicardial coronary capillary hemodynamics during reactive hyperemia and during adenosine administration by intravital video microscopy. AB - Using high-resolution intravital charge-coupled device video microscopy, we visualized the epicardial capillary network of the beating canine heart in vivo to elucidate its functional role under control conditions, during reactive hyperemia (RH), and during intracoronary adenosine administration. The pencil lens video-microscope probe was placed over capillaries fed by the left anterior descending artery in atrioventricular-blocked hearts of open-chest, anesthetized dogs paced at 60-90 beats/min (n = 17). In individual capillaries under control conditions, red blood cell flow was predominant during systole or diastole, indicating that the watershed between diastolic arterial and systolic venous flows is located within the capillaries. Capillary flow increased during RH and reached a peak flow velocity (2.1 +/- 0.6 mm/s), twice as high as control (1.2 +/ 0.5 mm/s), with enhancement of intercapillary cross-connection flow and enlargement of diameter (by 17%). With adenosine, capillary flow velocity significantly increased (1.8 +/- 0.7 mm/s). However, the increase in volumetric capillary flow with adenosine estimated from red blood cell velocity and diameter was less than the increase in arterial flow, whereas that during RH was nearly equivalent to the increase in arterial flow. There was a time lag of approximately 1.5 s for refilling of capillaries during RH, indicating their function as capacitance vessels. In conclusion, the coronary capillary network functions as 1) the major watershed between diastolic-dominant arterial and systolic-dominant venous flows, 2) a capacitor, and 3) a significant local flow amplifier and homogenizer of blood supply during RH, but with adenosine the increase in capillary flow velocity was less than the increase in arterial flow. PMID- 15345480 TI - PPAR-gamma agonist rosiglitazone ameliorates ventricular dysfunction in experimental chronic mitral regurgitation. AB - Previously we reported that the beneficial effects of beta-adrenergic blockade in chronic mitral regurgitation (MR) were in part due to induction of bradycardia, which obviously affects myocardial energy requirements. From this observation we hypothesized that part of the pathophysiology of MR may involve faulty energy substrate utilization, which in turn might lead to potentially harmful lipid accumulation as observed in other models of heart failure. To explore this hypothesis, we measured triglyceride accumulation in the myocardia of dogs with chronic MR and then attempted to enhance myocardial metabolism by chronic administration of the peroxisome proliferator-activated receptor (PPAR)-gamma agonist rosiglitazone. Cardiac tissues were obtained from three groups of dogs that included control animals, dogs with MR for 3 mo without treatment, and dogs with MR for 6 mo that were treated with rosiglitazone (8 mg/day) for the last 3 mo of observation. Hemodynamics and contractile function (end-systolic stress strain relationship, as measured by K index) were assessed at baseline, 3 mo of MR, and 6 mo of MR (3 mo of the treatment). Lipid accumulation in MR (as indicated by oil red O staining score and TLC analysis) was marked and showed an inverse correlation with the left ventricular (LV) contractility. LV contractility was significantly restored after PPAR therapy (K index: therapy, 3.01 +/- 0.11*; 3 mo MR, 2.12 +/- 0.34; baseline, 4.01 +/- 0.29; ANOVA, P = 0.038; *P < 0.05 vs. 3 mo of MR). At the same time, therapy resulted in a marked reduction of intramyocyte lipid. We conclude that 1) chronic MR leads to intramyocyte myocardial lipid accumulation and contractile dysfunction, and 2) administration of the PPAR-gamma agonist rosiglitazone ameliorates MR-induced LV dysfunction accompanied by a decline in lipid content. PMID- 15345481 TI - Distinct cAMP signaling pathways differentially regulate alpha2C-adrenoceptor expression: role in serum induction in human arteriolar smooth muscle cells. AB - The physiological role of alpha(2)-adrenoceptors (alpha(2)-ARs) in cutaneous, arteriolar, vascular smooth muscle cells (VSMs) is to mediate cold-induced constriction. In VSMs cultured from human cutaneous arterioles, there is a selective increase in alpha(2C)-AR expression after serum stimulation. In the present study, we examined the cellular mechanisms contributing to this response. Serum induction of alpha(2C)-ARs was paralleled by increased expression of cyclooxygenase-2 (COX-2), increased release of prostaglandins, and increased intracellular concentration of cAMP. Inhibition of COX-2 by acetyl salicylic acid (1 mM), NS-398 (5 microM), or celecoxib (3 microM) abolished the increase in cAMP and markedly reduced alpha(2C)-AR induction in response to serum stimulation. The cAMP agonists, forskolin (10 microM), isoproterenol (10 microM), and cholera toxin (0.1 microg/ml) each dramatically increased expression of alpha(2C)-ARs in human cutaneous VSMs. The A-kinase inhibitor H-89 (2 microM) inhibited phosphorylation of cAMP response element binding protein, but not the increase in alpha(2C)-AR expression in response to these agonists. cAMP-dependent but A kinase independent signaling can involve activation of guanine nucleotide exchange factors for the GTP-binding protein, Rap. Indeed, pull-down assays demonstrated Rap1 activation by serum and forskolin in VSM. Transient transfections using alpha(2C)-AR promoter-luciferase reporter construct demonstrated that Rap1 increased reporter activity, whereas the A-kinase catalytic subunit decreased reporter activity. These results indicate that cAMP signaling can have dual effects in cutaneous VSMs:activation of alpha(2C)-AR transcription mediated by Rap1 GTPase and suppression mediated by A-kinase. The former effect predominates in serum-stimulated VSMs leading to a COX-2, cAMP, and Rap 1-dependent increase in alpha(2C)-AR expression. Such increased expression of alpha(2C)-ARs may contribute to enhanced cold-induced vasoconstriction and Raynaud's phenomenon. PMID- 15345482 TI - Proteomic identification of 3-nitrotyrosine-containing rat cardiac proteins: effects of biological aging. AB - Proteomic techniques were used to identify cardiac proteins from whole heart homogenate and heart mitochondria of Fisher 344/Brown Norway F1 rats, which suffer protein nitration as a consequence of biological aging. Soluble proteins from young (5 mo old) and old (26 mo old) animals were separated by one- and two dimensional gel electrophoresis. One- and two-dimensional Western blots with an anti-nitrotyrosine antibody show an age-related increase in the immunoresponse of a few specific proteins, which were identified by nanoelectrospray ionization tandem mass spectrometry (NSI-MS/MS). Complementary proteins were immunoprecipitated with an immobilized anti-nitrotyrosine antibody followed by NSI-MS/MS analysis. A total of 48 proteins were putatively identified. Among the identified proteins were alpha-enolase, alpha-aldolase, desmin, aconitate hydratase, methylmalonate semialdehyde dehydrogenase, 3-ketoacyl-CoA thiolase, acetyl-CoA acetyltransferase, GAPDH, malate dehydrogenase, creatine kinase, electron-transfer flavoprotein, manganese-superoxide dismutase, F1-ATPase, and the voltage-dependent anion channel. Some contaminating blood proteins including transferrin and fibrinogen beta-chain precursor showed increased levels of nitration as well. MS/MS analysis located nitration at Y105 of the electron transfer flavoprotein. Among the identified proteins, there are important enzymes responsible for energy production and metabolism as well as proteins involved in the structural integrity of the cells. Our results are consistent with age dependent increased oxidative stress and with free radical-dependent damage of proteins. Possibly the oxidative modifications of the identified proteins contribute to the age-dependent degeneration and functional decline of heart proteins. PMID- 15345483 TI - Morphine mimics the antiapoptotic effect of preconditioning via an Ins(1,4,5)P3 signaling pathway in rat ventricular myocytes. AB - Morphine has cardioprotective effects against ischemic-reperfusion injuries. This study investigates whether morphine could mimic the antiapoptotic effect of preconditioning using a model of cultured neonatal rat cardiomyocytes subjected to metabolic inhibition (MI). To quantify MI-induced apoptosis, DNA fragmentation and mitochondrial cytochrome c release levels were measured by ELISA. MI dependent DNA fragmentation was prevented by both Z-VAD-fmk (20 microM), a pan caspase inhibitor, and cyclosporine A (CsA; 5 microM), a mitochondrial pore transition blocker, added during MI (36% and 54% decrease, respectively). MI dependent cytochrome c release was not blocked by Z-VAD-fmk but was decreased (38%) by CsA during MI. Metabolic preconditioning (MIP) and preconditioning with morphine (1 microM) were also assessed. MI-dependent DNA fragmentation and cytochrome c release were prevented by MIP (40% and 45% decrease, respectively) and morphine (34% and 45%, respectively). The antiapoptotic effect of morphine was abolished by naloxone (10 nM), a nonselective opioid receptor antagonist, or xestospongin C (XeC, 400 nM), an inhibitor of inositol (1,4,5)-trisphosphate [Ins(1,4,5)P(3)]-mediated Ca(2+) release. Ca(2+) preconditioning, induced by increasing extracellular Ca(2+) from 1.8 to 3.3 mM, mimicked the antiapoptotic effect of morphine on DNA fragmentation (24% decrease) and cytochrome c release (57% decrease). This effect mediated by extracellular Ca(2+) was also abolished by XeC. Measurements of intracellular Ca(2+) concentration using fura-2 microspectrofluorimetry showed that morphine induces Ins(1,4,5)P(3)-dependent Ca(2+) transients abolished by 2-aminoethoxydiphenyl borate (2-APB), a cell permeable Ins(1,4,5)P(3) antagonist. These results suggest that morphine preconditioning prevents simulated ischemia-reperfusion-induced apoptosis via an Ins(1,4,5)P(3) signaling pathway in rat ventricular myocytes. PMID- 15345484 TI - Impact of combined NO and PG blockade on rapid vasodilation in a forearm mild-to moderate exercise transition in humans. AB - We tested the hypothesis that nitric oxide (NO) and prostaglandins (PGs) contribute to the rapid vasodilation that accompanies a transition from mild to moderate exercise. Nine healthy volunteers (2 women and 7 men) lay supine with forearm at heart level. Subjects were instrumented for continuous brachial artery infusion of saline (control condition) or combined infusion of N(G)-nitro-L arginine methyl ester (L-NAME) and ketorolac (drug condition) to inhibit NO synthase and cyclooxygenase, respectively. A step increase from 5 min of steady state mild (5.4 kg) rhythmic, dynamic forearm handgrip exercise (1 s of contraction followed by 2 s of relaxation) to moderate (10.9 kg) exercise for 30 s was performed. Steady-state forearm blood flow (FBF; Doppler ultrasound) and forearm vascular conductance (FVC) were attenuated in drug compared with saline (control) treatment: FBF = 196.8 +/- 30.8 vs. 281.4 +/- 34.3 ml/min and FVC = 179.3 +/- 29.4 vs. 277.8 +/- 34.8 ml.min(-1).100 mmHg(-1) (both P < 0.01). FBF and FVC increased from steady state after release of the initial contraction at the higher workload in saline and drug conditions: DeltaFBF = 72.4 +/- 8.7 and 52.9 +/- 7.8 ml/min, respectively, and DeltaFVC = 66.3 +/- 7.3 and 44.1 +/- 7.0 ml.min(-1).100 mmHg(-1), respectively (all P < 0.05). The percent DeltaFBF and DeltaFVC were not different during saline infusion or combined inhibition of NO and PGs: DeltaFBF = 27.2 +/- 3.1 and 28.1 +/- 3.8%, respectively (P = 0.78) and DeltaFVC = 25.7 +/- 3.2 and 26.0 +/- 4.0%, respectively (P = 0.94). The data suggest that NO and vasodilatory PGs are not obligatory for rapid vasodilation at the onset of a step increase from mild- to moderate-intensity forearm exercise. Additional vasodilatory mechanisms not dependent on NO and PG release contribute to the immediate and early increase in blood flow in an exercise-to-exercise transition. PMID- 15345485 TI - Effects of menstrual cycle and oral contraceptive use on calf venous compliance. AB - Numerous studies have shown that the female sex hormones estrogen and progesterone have multiple effects on the vasculature. Thus our goal was to investigate the effects of estrogen and progesterone on calf venous compliance by looking for cyclic changes during the early follicular, ovulatory, and midluteal phases of the menstrual cycle and during high and low hormone phases of oral contraceptive use. Additionally, we wanted to compare the venous compliance of normally menstruating women, oral contraceptive users, and men. We studied eight normally menstruating women (23 +/- 1 yr of age) during the early follicular, ovulatory, and midluteal phases of the menstrual cycle. Nine triphasic oral contraceptive users (21 +/- 1 yr of age) were studied during weeks of high and low hormone concentrations. Eight men (23 +/- 1 yr of age) were studied twice within 2-4 wk. With the use of venous occlusion plethysmography with mercury in Silastic strain gauges, lower limb venous compliance was measured by inflating a venous collection cuff that was placed on the thigh to 60 mmHg for 8 min and then reducing the pressure to 0 mmHg at a rate of 1 mmHg/s. Venous compliance was calculated as the derivative of the pressure-volume curves. There were no differences between early follicular, ovulatory, and midluteal phases of the menstrual cycle or between high and low hormone phases of oral contraceptive use (P > 0.05). Male venous compliance was significantly greater than in normally menstruating women (P < 0.001) and oral contraceptive users (P < 0.002). These data support a sex difference but also suggest that venous compliance does not change with menstrual cycle phase or during the course of oral contraceptive use. PMID- 15345486 TI - Role of hydrogen peroxide in ACh-induced dilation of human submucosal intestinal microvessels. AB - The endothelium plays an important role in maintaining vascular homeostasis by synthesizing and releasing several mediators of vasodilation, which include prostacyclin (PGI(2)), nitric oxide, and endothelium-derived hyperpolarizing factor (EDHF). We have recently defined the role of nitric oxide and PGI(2) in the dilation of submucosal intestinal arterioles from patients with normal bowel function. However, significant endothelium-dependent dilator capacity to ACh remained after inhibiting both these mediators. The current study was designed to examine the potential role of EDHF in human intestinal submucosal arterioles. ACh elicited endothelium-dependent relaxation in the presence of inhibitors of nitric oxide synthase and cyclooxygenase (23 +/- 10%, n = 6). This ACh-induced relaxation was inhibited and converted to constriction by catalase (-53 +/- 10%, n = 6) or KCl (-30 +/- 3%, n = 7), whereas 17-octadecynoic acid and 6-(2 propargylloxyphenyl) hexanoic acid, two inhibitors of cytochrome P450 monooxygenase, had no significant effect (3 +/- 1% and 20 +/- 8%, n = 5, respectively). Exogenous H(2)O(2) elicited dose-dependent relaxation of intact microvessels (52 +/- 10%, n = 7) but caused frank vasoconstriction in arterioles denuded of endothelium (-73 +/- 8%, n = 7). ACh markedly increased the dichlorofluorescein fluorescence in intact arterioles in the presence of nitric oxide synthase and cyclooxygenase inhibitors compared with control and compared with catalase-treated microvessels (363.6 +/- 49, 218.8 +/- 10.6, 221.9 +/- 27.9, respectively, P < 0.05 ANOVA, n = 5 arbitrary units). No changes in the dichlorofluorescein fluorescence were recorded in vessels treated with ACh alone. These results indicate that endothelial production of H(2)O(2) occurs in response to ACh in human gut mucosal arterioles but that H(2)O(2) is not an EDHF in this tissue. Rather, we speculate that it stimulates the release of a chemically distinct EDHF. PMID- 15345487 TI - Effects of PPAR-gamma ligands on vascular smooth muscle marker expression in hypertensive and normal arteries. AB - Having previously demonstrated that glucose transporter-4 (GLUT4) expression was reduced in aortas and carotid arteries of deoxycorticosterone acetate (DOCA) salt hypertensive rats, we hypothesized that troglitazone (TG), through activation of peroxisome proliferator-activated receptor-gamma (PPAR-gamma), would stabilize GLUT4 expression and possibly preserve the differentiated phenotype in vascular smooth muscle cells. In DOCA salt-hypertensive rats treated with TG (100 mg/day), there was a significant (P < 0.001) decrease in systolic blood pressure (BP; 149.9 +/- 4.4 mmHg) compared with the untreated DOCA salt-hypertensive rats (202.2 +/- 10.34 mmHg). Separate trials with rosiglitazone (RS; 3 mg/day) demonstrated a significant (P < 0.001) decrease in BP (DOCA salt, 164.2 +/- 9.8 vs. DOCA-RS, 124.9 +/- 3.7 mmHg) comparable to that with TG. Expression of GLUT4, h-caldesmon, and smooth muscle myosin heavy chain SM2 was significantly decreased in aortas of DOCA salt-hypertensive rats and was reversed by TG to levels similar to those in aortas of sham-treated rats. TG (50 microM) induced GLUT4 and h caldesmon expression in 24-h culture of explanted carotid arteries of DOCA salt hypertensive rats, and the endogenous PPAR-gamma ligand 15-deoxy-Delta(12-14) prostaglandin J(2) (PGJ(2); 20 microM) and TG (50 microM) similarly increased GLUT4, h-caldesmon, and SM2 protein expression in explanted aortas. The expression of activated, phosphorylated Akt was increased by PGJ(2) and TG with no significant effect on total Akt levels. Inhibition of phosphorylated Akt expression using the phosphatidylinositol 3-kinase inhibitor LY-294002 (16 microM) abrogated the increased expression of h-caldesmon and SM2. These data demonstrate that PPAR-gamma agonists maintain or induce expression of markers of the contractile phenotype independently of their effects on hypertension, and that this effect may be mediated through activation of phosphatidylinositol 3 kinase/Akt. PMID- 15345488 TI - Angiotensin II-induced hypertrophy is potentiated in mice overexpressing p22phox in vascular smooth muscle. AB - Increased reactive oxygen species (ROS) are implicated in several vascular pathologies associated with vascular smooth muscle hypertrophy. In the current studies, we utilized transgenic (Tg) mice (Tg(p22smc)) that overexpress the p22(phox) subunit of NAD(P)H oxidase selectively in smooth muscle. These mice have a twofold increase in aortic p22(phox) expression and H(2)O(2) production and thus provide an excellent in vivo model in which to assess the effects of increased ROS generation on vascular smooth muscle cell (VSMC) function. We tested the hypothesis that overexpression of VSMC p22(phox) potentiates angiotensin II (ANG II)-induced vascular hypertrophy. Male Tg(p22smc) mice and negative littermate controls were infused with either ANG II or saline for 13 days. Baseline blood pressure was not different between control and Tg(p22smc) mice. ANG II significantly increased blood pressure in both groups, with this increase being slightly exacerbated in the Tg(p22smc) mice. Baseline aortic wall thickness and cross-sectional wall area were not different between control and Tg(p22smc) mice. Importantly, the ANG II-induced increase in both parameters was significantly greater in the Tg(p22smc) mice compared with control mice. To confirm that this potentiation of vascular hypertrophy was due to increased ROS levels, additional groups of mice were coinfused with ebselen. This treatment prevented the exacerbation of hypertrophy in Tg(p22smc) mice receiving ANG II. These data suggest that although increased availability of NAD(P)H oxidase derived ROS is not a sufficient stimulus for hypertrophy, it does potentiate ANG II-induced vascular hypertrophy, making ROS an excellent target for intervention aimed at reducing medial thickening in vivo. PMID- 15345489 TI - Cytosolic NADPH may regulate differences in basal Nox oxidase-derived superoxide generation in bovine coronary and pulmonary arteries. AB - Because systems controlled by basal NAD(P)H oxidase activity appear to contribute to differences in responses of endothelium-removed bovine coronary (BCA) and pulmonary (BPA) arteries to hypoxia, we characterized the Nox oxidases activities present in these vascular segments and how cytosolic NAD(P)H redox systems could be controlling oxidase activity. BPA generated approximately 60-80% more lucigenin (5 microM) chemiluminescence detectable superoxide than BCA. Apocynin (10 microM), a NAD(P)H oxidase inhibitor, and 6-aminonicotinamide (1 mM), a pentose phosphate inhibitor (PPP), both attenuated (approximately by 50-70%) superoxide detected in BPA and BCA. There was no significant difference in the expression of Nox2 or Nox4 mRNA or protein detected by Western blot analysis. NADPH and NADH increased superoxide in homogenates and isolated microsomal membrane fractions in a manner consistent with BPA and BCA having similar levels of oxidase activity. BPA had 4.2-fold higher levels of NADPH than BCA. The activity and protein levels of glucose-6-phosphate dehydrogenase (G6PD), the rate limiting PPP enzyme generating cytosolic NADPH, were 1.5-fold higher in BPA than BCA. Thus BPA differ from BCA in that they have higher levels of G6PD activity, NADPH, and superoxide. Because both arteries have similar levels of Nox expression and activity, elevated levels of cytosolic NADPH may contribute to increased superoxide in BPA. PMID- 15345490 TI - Effects of sodium nitroprusside in aortic stenosis associated with severe heart failure: pressure-volume loop analysis using a numerical model. AB - In the recently published clinical study [Use of Nitroprusside in Left Ventricular Dysfunction and Obstructive Aortic Valve Disease (UNLOAD)], sodium nitroprusside (SNP) improved cardiac function in patients with severe aortic stenosis (AS) and left ventricular (LV) systolic dysfunction. We explored the possible mechanisms of these findings using a series of numerical simulations. A closed-loop lumped parameters model that consists of 24 differential equations relating pressure and flow throughout the circulation was used to analyze the effects of varying hemodynamic conditions in AS. Hemodynamic data from UNLOAD study subjects were used to construct the initial simulation. Systemic vascular resistance (SVR), heart rate, and aortic valve area were directly entered into the model while end-systolic and end-diastolic pressure-volume (P-V) relationships were adjusted using previously published data to match modeled and observed end-systolic and end-diastolic pressures and volumes. Initial simulation of SNP treatment by a reduction of SVR was not adequate. To obtain realistic model hemodynamics that reliably reproduce SNP treatment effects, we performed a series of simulations while simultaneously changing end-systolic elastance (E(es)), end-systolic volume at zero pressure (V(0)), and diastolic P-V shift. Our data indicate that either an E(es) increase or V(0) decrease is necessary to obtain realistic model hemodynamics. In five patients, we corroborated our findings by using the model to duplicate individual P-V loops obtained before and during SNP treatment. In conclusion, using a numerical model, we identified ventricular function parameters that are responsible for improved hemodynamics during SNP infusion in AS with LV dysfunction. PMID- 15345491 TI - Why is flow-mediated dilation dependent on arterial size? Assessment of the shear stimulus using phase-contrast magnetic resonance imaging. AB - Flow-mediated dilation (FMD) is strongly dependent on arterial size, but the reasons for this phenomenon are poorly understood. We have previously shown that FMD is greater in small brachial arteries because the shear stress stimulus is greater in small brachial arteries. However, it is unclear why the shear stimulus is greater in small arteries. Furthermore, this relationship has not been investigated in other, differently sized arterial beds. Postischemic systolic shear stress and resulting FMD were evaluated in the brachial and femoral arteries of 24 young, healthy adults using phase-contrast magnetic resonance imaging. Arterial shear and radius were calculated from the velocity profile via a best-fit parabola before and after occlusion. Summing the velocity pixels provided hyperemic systolic flow. FMD was proportional to hyperemic shear in the brachial and femoral arteries (P < 0.0001, r = 0.60). Hyperemic systolic flow was proportional to radius2 (P < 0.0001, r = 0.93). Applying this relationship to the Poiseuille equation (shear is proportional to flow/radius3) shows that hyperemic shear is proportional to radius2/radius3 and, therefore, explains why hyperemic shear is proportional to 1/radius. We conclude that FMD is proportional to hyperemic systolic shear stress in both the brachial and the femoral arteries. The hyperemic shear stimulus for FMD is greater in small arteries due to the dependence of postischemic systolic flow on radius squared. Therefore, greater FMD in small arteries does not necessarily reflect better conduit artery endothelial function. Evaluating the shear stimulus using phase-contrast magnetic resonance imaging enhances the understanding of mechanisms underlying FMD. PMID- 15345492 TI - Contrasting effects of ischemia on the kinetics of membrane voltage and intracellular calcium transient underlie electrical alternans. AB - Repolarization alternans has been considered a strong marker of electrical instability. The objective of this study was to investigate the hypothesis that ischemia-induced contrasting effects on the kinetics of membrane voltage and intracellular calcium transient (Ca(i)T) can explain the vulnerability of the ischemic heart to repolarization alternans. Ischemia-induced changes in action potential (AP) and Ca(i)T resulting in alternans were investigated in perfused Langendorff guinea pig hearts subjected to 10-15 min of global no-flow ischemia followed by 10-15 min of reperfusion. The heart was stained with 100 microl of rhod-2 AM and 25 microl of RH-237, and AP and Ca(i)T were simultaneously recorded with an optical mapping system of two 16 x 16 photodiode arrays. Ischemia was associated with shortening of AP duration (D) but delayed upstroke, broadening of peak, and slowed decay of Ca(i)T resulting in a significant increase of Ca(i)T-D. The changes in APD were spatially heterogeneous in contrast to a more spatially homogeneous lengthening of Ca(i)T-D. Ca(i)T alternans could be consistently induced with the introduction of a shorter cycle when the upstroke of the AP occurred before complete relaxation of the previous Ca(i)T and generated a reduced Ca(i)T. However, alternans of Ca(i)T was not necessarily associated with alternans of APD, and this was correlated with the degree of spatially heterogeneous shortening of APD. Sites with less shortening of APD developed alternans of both Ca(i)T and APD, whereas sites with greater shortening of APD could develop a similar degree of Ca(i)T alternans but slight or no APD alternans. This resulted in significant spatial dispersion of APD. The study shows that the contrasting effects of ischemia on the duration of AP and Ca(i)T and, in particular, on their spatial distribution explain the vulnerability of ischemic heart to alternans and the increased dispersion of repolarization during alternans. PMID- 15345493 TI - Sodium and potassium handling by the aldosterone-sensitive distal nephron: the pivotal role of the distal and connecting tubule. AB - Sodium reabsorption and potassium secretion in the distal convoluted tubule and in the connecting tubule can maintain the homeostasis of the body, especially when dietary sodium intake is high and potassium intake is low. Under these conditions, a large proportion of the aldosterone-regulated sodium and potassium transport would occur in these nephron segments before the tubular fluid reaches the collecting duct. The differences between these two segments and the collecting duct would be more quantitative than qualitative. The collecting duct would come into play when the upstream segments are overloaded by a primary genetic defect that affects sodium and/or potassium transport or by a diet that is exceedingly poor in sodium and rich in potassium. It is likely that the homeostatic role of the distal convoluted and connecting tubules, which are technically difficult to study, has been underestimated, whereas the role of the more easily accessible collecting duct may have been overemphasized. PMID- 15345494 TI - Ammonia production and secretion by S3 proximal tubule segments from acidotic mice: role of ANG II. AB - ANG II has potent effects on ammonia production and secretion rates by the proximal tubule and is found in substantial concentrations in the lumen of the proximal tubule in vivo. Because our previous studies demonstrated that acid loading enhanced the stimulatory effects of ANG II on ammonia production and secretion by S2 proximal tubule segments, we examined the effect of ANG II on ammonia production and secretion by isolated, perfused S3 segments from nonacidotic control mice and acidotic mice given NH4Cl for 7 days. In the absence of ANG II, ammonia production and secretion rates were no different in S3 segments from acidotic and control mice. By contrast, when ANG II was present in the luminal perfusion solution, ammonia production and secretion rates were stimulated, in a losartan-inhibitable manner, to a greater extent in S3 segments from acidotic mice. Ammonia secretion rates in S3 segments were largely inhibited by perfusion with a low-sodium solution containing amiloride in the presence or absence of ANG II. These results demonstrated that isolated, perfused mouse S3 proximal tubule segments produce and secrete ammonia, that NH4Cl-induced acidosis does not affect the basal rates of ammonia production and secretion, and that ANG II, added to the luminal fluid, stimulates ammonia production and secretion to a greater extent in S3 segments from acidotic mice. These findings suggest that S3 segments, in the presence of ANG II, can contribute to the enhanced renal excretion that occurs with acid loading. PMID- 15345495 TI - Effect of muscimol and L-NAME in the PVN on the RSNA response to volume expansion in conscious rabbits. AB - In the present study, we have investigated whether the hypothalamic paraventricular nucleus (PVN) contributed to the reflex reduction in renal sympathetic nerve activity (RSNA) normally elicited by volume expansion in the conscious rabbit. RSNA was monitored after volume expansion (Dextran 70, 2 ml/min for 30 min) in animals microinjected into, and outside, the PVN with muscimol (10 nmol), to acutely inhibit neuronal function. Because nitric oxide within the PVN inhibits RSNA, we also examined the effect of NG-nitro-L-arginine methyl ester (L NAME; 20 nmol) to block nitric oxide synthase. Compared with vehicle, the reduction in RSNA elicited by volume expansion was abolished by injection of muscimol into the PVN. The effect was specific to the PVN because microinjections of muscimol outside the PVN had no effect on the response. L-NAME microinjected into or outside the PVN had no effect on the RSNA response. The findings suggest that the PVN is essential in the central pathways mediating the renal sympathetic nerve response elicited by elevations in plasma volume but that nitric oxide does not play a major role. PMID- 15345496 TI - Extracellular Ca2+ regulates the stimulation of Na+ transport in A6 renal epithelia. AB - We investigated the involvement of intracellular and extracellular Ca2+ in the stimulation of Na+ transport during hyposmotic treatment of A6 renal epithelia. A sudden osmotic decrease elicits a biphasic stimulation of Na+ transport, recorded as increase in amiloride-sensitive short-circuit current (Isc) from 3.4 +/- 0.4 to 24.0 +/- 1.3 microA/cm2 (n = 6). Changes in intracellular Ca2+ concentration ([Ca2+]i) were prevented by blocking basolateral Ca2+ entry with Mg2+ and emptying the intracellular Ca2+ stores before the hyposmotic challenge. This treatment did not noticeably affect the hypotonicity-induced stimulation of Isc. However, the absence of extracellular Ca2+ severely attenuated Na+ transport stimulation by the hyposmotic shock, and Isc merely increased from 2.2 +/- 0.3 to 4.8 +/- 0.7 microA/cm2. Interestingly, several agonists of the Ca2+-sensing receptor, Mg2+ (2 mM), Gd3+ (0.1 mM), neomycin (0.1 mM), and spermine (1 mM) were able to substitute for extracellular Ca2+. When added to the basolateral solution, these agents restored the stimulatory effect of the hyposmotic solutions on Isc in the absence of extracellular Ca2+ to levels that were comparable to control conditions. None of the above-mentioned agonists induced a change in [Ca2+]i. Quinacrine, an inhibitor of PLA2, overruled the effect of the agonists on Na+ transport. In conclusion, we suggest that a Ca2+-sensing receptor in A6 epithelia mediates the stimulation of Na+ transport without the interference of changes in [Ca2+]i. PMID- 15345497 TI - Expression of resistance markers to methotrexate predicts clinical improvement in patients with rheumatoid arthritis. AB - BACKGROUND: Methotrexate is transported into the cell by the reduced folate carrier (RFC) and out of the cell by members of the multidrug resistance protein family (MRP). Transport proteins may affect the therapeutic efficacy of this drug in patients with rheumatoid arthritis. OBJECTIVE: To investigate the potential benefit of the presence of RFC and the absence of functional MRP for the efficacy of methotrexate treatment. METHODS: The study involved 163 patients (116 female, 47 male; mean age 59.5 years) on methotrexate (mean weekly dose 12.2 mg). RFC was determined using reverse transcriptase polymerase chain reaction, and MRP function by flow cytometry, using a calcein acetoxymethylesther/probenecid assay. Clinical response to methotrexate was evaluated by the EULAR response criteria and the ACR 20% improvement criteria. The clinical data were obtained at the beginning of methotrexate treatment and at the time of blood sampling during ongoing therapy. Patients were divided into four groups according to the presence (+) or absence (-) of RFC and functional (f) MRP. RESULTS: fMRP+/RFC+ and fMRP /RFC- patients more often had good EULAR response rates (60%, p = 0.014, and 53%, p = 0.035, respectively) in comparison with the fMRP-/RFC+ group (29%); fMRP+/RFC patients had a low frequency of good disease activity responses. CONCLUSIONS: Absence of fMRP plus presence of RFC did not prove to be related to beneficial effects of methotrexate, but the lack or the presence of both fMRP and RFC led to a significantly better therapeutic outcome. Determination of these markers may predict responsiveness to methotrexate. PMID- 15345498 TI - Outcomes of a multicentre randomised clinical trial of etanercept to treat ankylosing spondylitis. AB - OBJECTIVE: A double blind, randomised, placebo controlled study to evaluate the safety and efficacy of etanercept to treat adult patients with ankylosing spondylitis (AS). METHODS: Adult patients with AS at 14 European sites were randomly assigned to 25 mg injections of etanercept or placebo twice weekly for 12 weeks. The primary efficacy end point was an improvement of at least 20% in patient reported symptoms, based on the multicomponent Assessments in Ankylosing Spondylitis (ASAS) response criteria (ASAS 20). Secondary end points included ASAS 50 and ASAS 70 responses and improved scores on individual components of ASAS, the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), acute phase reactants, and spinal mobility tests. Safety was evaluated during scheduled visits. RESULTS: Of 84 patients enrolled, 45 received etanercept and 39 received placebo. Significantly more etanercept patients than placebo patients responded at the ASAS 20 level as early as week 2, and sustained differences were evident up to week 12. Significantly more etanercept patients reported ASAS 50 responses at all times and ASAS 70 responses at weeks 2, 4, and 8; reported lower composite and fatigue BASDAI scores; had lower acute phase reactant levels; and had improved spinal flexion. Etanercept was well tolerated. Most adverse events were mild to moderate; the only between-group difference was injection site reactions, which occurred significantly more often in etanercept patients. CONCLUSIONS: Etanercept is a well tolerated and effective treatment for reducing clinical symptoms and signs of AS. PMID- 15345499 TI - Galectin-3 is induced in rheumatoid arthritis synovial fibroblasts after adhesion to cartilage oligomeric matrix protein. AB - BACKGROUND: Galectin-3 is expressed in the synovial tissue of patients with rheumatoid arthritis (RA), particularly at sites of joint destruction. OBJECTIVE: To explore the possibilities that galectin-3 is induced either by proinflammatory cytokines or by adhesion to cartilage components. METHODS: Cell culture plates were coated with fibronectin, collagens I-VI, or cartilage oligomeric matrix protein (COMP), and the suspended cells were then added. The medium was changed after 1 hour at 37 degrees C. Adherent cells were further incubated for 18 hours in the presence or absence of tumour necrosis factor alpha (TNF alpha) or interleukin 1 beta. Cells were pretreated with murine IgG1, anti-CD29, -CD51, CD61 (integrins), or -CD3 monoclonal antibodies and transferred to culture plates coated with COMP. Adherent cells were counted by light microscopy. The expression of intracellular galectin-3, or cell surface CD29, CD51, and CD61 was determined by flow cytometry before and after adhesion. RESULTS: Four times more RA synovial fibroblasts (SF) than osteoarthritis SF adhered to COMP. RA SF presented more cell surface integrins, and monoclonal antibodies against CD51 inhibited the adhesion to COMP by 80%. TNF alpha reduced the expression of CD61 and the adhesion to COMP, but did not reverse the adhesion once it had taken place. The adhesion of RA SF to COMP was found to increase the intracellular level of galectin-3. In contrast, intracellular galectin-3 decreased after exposure to TNF alpha. CONCLUSION: The increase of galectin-3 occurs after adhesion to COMP, and the alpha V beta 3 receptor (CD51/CD61) has a pivotal role in this process. PMID- 15345500 TI - A randomised, placebo controlled, comparative trial of the gastrointestinal safety and efficacy of AZD3582 versus naproxen in osteoarthritis. AB - OBJECTIVE: To evaluate the gastrointestinal safety and efficacy of the COX inhibiting nitric oxide donator AZD3582 in patients with hip or knee osteoarthritis. METHODS: 970 patients were randomised (7:7:2) to AZD3582 750 mg twice daily, naproxen 500 mg twice daily, or placebo twice daily in a double blind study. The primary end point was the six week incidence of endoscopic gastroduodenal ulcers (diameter > or =3 mm). Overall damage measured on the Lanza scale was a secondary end point. Safety and tolerability assessments included endoscopic upper gastrointestinal erosions and the gastrointestinal symptom rating scale (GSRS). Efficacy was primarily assessed by WOMAC. RESULTS: The incidence of ulcers with AZD3582 was 9.7% and with naproxen 13.7% (p = 0.07, NS), v 0% on placebo. The incidence of Lanza scores >2 was higher with naproxen (43.7%) than with AZD3582 (32.2%) (p<0.001). Compared with baseline, significantly fewer ulcers and erosions developed in stomach and stomach/duodenum combined, and fewer erosions developed in stomach, duodenum, and both combined on AZD3582 than on naproxen. GSRS reflux and abdominal pain subscale scores were lower for AZD3582 than for naproxen but there was no difference for indigestion, constipation, and diarrhoea. AZD3582 was as effective as naproxen at improving WOMAC scores. Both agents were well tolerated, with no significant effects on blood pressure. CONCLUSIONS: At doses with similar efficacy in relieving osteoarthritis symptoms, the primary end point of six week endoscopic gastroduodenal ulcer incidence was not significantly different between AZD3582 and naproxen. Most secondary endoscopic gastrointestinal end points favoured AZD3582. PMID- 15345501 TI - Therapeutic drug monitoring of A77 1726, the active metabolite of leflunomide: serum concentrations predict response to treatment in patients with rheumatoid arthritis. AB - BACKGROUND: Leflunomide is the prodrug of the disease modifying antirheumatic metabolite A77 1726. More than 50% of patients withdraw from leflunomide treatment within one year, mainly because of adverse drug reactions. Therapeutic drug monitoring of A77 1726 may be useful in predicting the efficacy of leflunomide treatment. OBJECTIVE: To study the relation between A77 1726 steady state serum concentrations and disease activity using the 28 joint (DAS28) response. METHODS: Outpatients with rheumatoid arthritis on a stable leflunomide dose for >4 months were included. DAS28 score and adverse drug reactions were recorded. Blood samples were taken for determination of A77 1726 concentrations. The primary end point was the relation of serum A77 1726 concentrations with DAS28 response category. RESULTS: Serum A77 1726 concentrations were determined in 52 patients. A receiver operating characteristic (ROC) curve showed an area under the curve (AUC) of 0.73 (95% confidence interval, 0.54 to 0.93) (p<0.05). The sensitivity exceeded 99% at concentrations below 16 mg/l. DAS28 values at the point of sampling showed no relation with A77 1726 concentrations (AUC of the ROC curve = 0.50 (0.33 to 0.67) (NS)). CONCLUSIONS: A77 1726 steady state serum concentrations show a relation with DAS28 response. Determination of serum A77 1726 concentrations in patients with insufficient response to treatment may help when decisions have to be made about continuation of treatment or dose adjustment. PMID- 15345502 TI - Preliminary results of safety and efficacy of the interleukin 1 receptor antagonist anakinra in patients with severe lupus arthritis. AB - BACKGROUND: Joint involvement occurs in most patients with systemic lupus erythematosus (SLE), and severe lupus arthritis is often refractory to conventional treatments. Anakinra is used in the treatment of rheumatoid arthritis, but its therapeutic potential has not been proved in patients with SLE. OBJECTIVE: To determine the safety/tolerability and efficacy of anakinra in patients with SLE with leading joint involvement. METHODS: In patients with SLE with active polyarthritis and no other uncontrolled systemic/organ manifestations, 100 mg/day anakinra was self administered subcutaneously for 3 months. Disease activity was assessed by VAS, number of swollen/tender joints, ECLAM score, and serological and immunological measures. RESULTS: Four patients with SLE were studied; anakinra was safe in all four patients and no drug related serious adverse events occurred. A subjective benefit was seen in all patients and a trend towards better activity measures after 4 weeks. After an initial response, one patient left the study because of an arthritic flare after 6 weeks. CONCLUSION: In this study anakinra was apparently safe and well tolerated and led to clinical and serological improvement. Anakinra might be an interesting alternative in individual patients with lupus arthritis not responding to conventional treatments. PMID- 15345503 TI - Methotrexate ameliorates T cell dependent autoimmune arthritis and encephalomyelitis but not antibody induced or fibroblast induced arthritis. AB - OBJECTIVE: To investigate the mode of action of methotrexate (MTX) in different types of models for rheumatoid arthritis (RA) and multiple sclerosis (MS). METHODS: Models for RA and MS were selected known to have different pathogenesis- that is, fibroblast induced arthritis in SCID mice, collagen induced arthritis (CIA), anticollagen II antibody induced arthritis (CAIA), and experimental autoimmune encephalomyelitis (EAE) in (Balb/c x B10.Q)F1 and B10.Q mice, and Pristane induced arthritis in DA rats (PIA). The MTX treatment was started 1 day after the onset of disease and continued for 14 days to compare effects on the different models. RESULTS: All models known to be critically dependent on T cell activation (CIA, PIA, and EAE) were effectively down regulated by titrated doses of MTX. In contrast, no effects were seen on fibroblast induced arthritis or CAIA. No effects were seen on the levels of anticollagen II antibodies in the CIA experiment. CONCLUSION: The data show that MTX has strong ameliorative effect on both classical models of RA, like CIA and PIA, but also on a model for MS, EAE. It also suggests that MTX operates only in diseases which are preceded by, and dependent on, T cell activation. A comparison of CAIA and CIA suggested that MTX operates independently of arthritogenic antibodies. These results demonstrate that different animal models reflect the complexity of the corresponding human diseases and suggest that several models should be used for effective screening of new therapeutic agents. PMID- 15345504 TI - Interleukin 10 promoter microsatellite polymorphisms are associated with response to long term treatment with etanercept in patients with rheumatoid arthritis. AB - OBJECTIVES: To analyse the association of interleukin 10 (IL10) promoter polymorphisms, which have been shown to be related to IL10 secretion capacity, with the response to long term treatment with etanercept in patients with rheumatoid arthritis (RA). METHODS: Fifty patients with active RA were treated for up to 4 years (median 39 months, range 3-52) with stable doses of etanercept as monotherapy. Treatment response was assessed as defined by the EULAR criteria in an intention to treat analysis, with the last observation carried forward. IL10 promoter microsatellite polymorphisms IL10.R and IL10.G were genotyped by fragment length analysis in patients and 189 healthy controls matched for ethnicity, age, and sex. Haplotypes were reconstructed using a method based on bayesian, coalescent theory with the PHASE software. RESULTS: IL10 microsatellite polymorphisms were not associated with susceptibility to RA. When patients with good treatment response (n = 25) were compared with patients with moderate (n = 17) or no response (n = 8), a significantly different distribution of the prevailing alleles R2, R3 and G9, G13, respectively, became evident. Good treatment response was associated with carriage of the R3 allele or R3-G9 haplotype, whereas the allele G13 and the haplotype R2-G13 predominated in patients with moderate or no response. CONCLUSION: Genotyping of the IL10 promoter microsatellites may be useful in predicting the clinical response to etanercept in patients with RA. The high prevalence of the presumptive IL10 low producer allele R3 in patients with a favourable response suggests that IL10 promotes disease activity in RA under the specific condition of tumour necrosis factor antagonism. PMID- 15345505 TI - Asynchronous shear stress and circumferential strain reduces endothelial NO synthase and cyclooxygenase-2 but induces endothelin-1 gene expression in endothelial cells. AB - OBJECTIVE: Endothelium-derived vasoactive agents NO, endothelin-1 (ET-1), and prostacyclin (PGI2) not only regulate vascular tone but also influence atherogenic processes, including smooth muscle migration and proliferation, as well as monocyte and platelet adhesion. Complex hemodynamics characterized by the temporal phase angle between mechanical factors circumferential strain and wall shear stress (stress phase angle [SPA]) have been implicated in regions prone to pathologic development, such as atherosclerosis and intimal hyperplasia, in coronary and peripheral arteries where the mechanical forces are highly asynchronous (SPA=-180 degrees ). We determined the gene expression of endothelial NO synthase (eNOS), ET-1, and cyclooxygenase-2 (COX-2) affected by asynchronous hemodynamics (SPA=-180 degrees ) relative to normal hemodynamics (SPA=0 degrees ) in bovine aortic endothelial cells. METHODS AND RESULTS: Quantitative competitive RT-PCR analysis showed that eNOS production (at 5 and 12 hours) and COX-2 production (at 5 hours) were reduced at the gene expression level by asynchronous hemodynamics (SPA=-180 degrees) compared with synchronous hemodynamics (SPA=0 degrees ), whereas ET-1 exhibited an opposite trend (at 5 and 12 hours). NO, ET-1, and PGI2 secretion followed their respective gene expression profiles after 5 and 12 hours. CONCLUSIONS: Together, these data suggest that highly asynchronous mechanical force patterns (SPA=-180 degrees ) can elicit proatherogenic vasoactive responses in endothelial cells at the gene expression level, indicating a novel mechanism that induces cardiovascular pathology. PMID- 15345506 TI - Genetic influence on inflammation variables in the elderly. AB - BACKGROUND: Inflammation variables (C-reactive protein [CRP], fibrinogen, and soluble intercellular adhesion molecule-1 [sICAM-1]) have been identified as risk factors for cardiovascular disease. It is still not known how much the regulation of inflammatory risk factors is determined by genetic factors, and the aim of this study was to determine the heritability of these inflammation variables and of the acute phase regulating cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) at older ages. METHODS AND RESULTS: The heritability of CRP, fibrinogen, sICAM-1, IL-6, and TNF-alpha was determined in a twin study consisting of 129 monozygotic twin pairs and 153 dizygotic same-sex twins aged 73 to 94 years who participated in the Longitudinal Study of Aging of Danish Twins. Furthermore, we determined the influence of selected genetic polymorphisms on the plasma level variations. Genetic factors accounted for 20% to 55% of the variation in plasma levels of the inflammation variables. The highest heritability was found for sICAM-1. The genetic polymorphisms we studied explained only a small, insignificant part of the heritability. CONCLUSIONS: This study in elderly twins provides evidence for a substantial genetic component of inflammatory cardiovascular risk factors among the elderly. PMID- 15345507 TI - Beta3-integrin regulates vascular endothelial growth factor-A-dependent permeability. AB - OBJECTIVE: Beta3-integrin deficiency has been implicated in increasing levels of Flk-1 expression on endothelial cells and enhancing vascular endothelial growth factor (VEGF)-induced angiogenesis. We determined the role of beta3-integrin in mediating VEGF-A-induced blood vessel permeability through Flk-1. METHODS AND RESULTS: Using the Miles assay, we demonstrated that VEGF-A-induced plasma leakage was enhanced in beta3-null mice when compared with wild-type controls. This was not caused by any changes in blood vessel structure (as detected by light or electron microscopy) or by changes in endothelial cell-cell adhesion proteins (as determined by Western blot analysis, flow cytometry, and immunofluorescence). Circulating levels of VEGF, baseline blood vessel leakage, and leakage in response to an acute inflammatory stimulus were identical in wild type and beta3-null mice. However, VEGF-A-induced leakage was abolished in beta3 null mice by the inhibition of Flk-1, indicating that the elevated levels of Flk 1 on beta3-null endothelial cells enhance VEGF-A-induced permeability. CONCLUSIONS: beta3-integrin-deficiency increases the sensitivity of endothelial cells to VEGF-A by elevating Flk-1 expression and, as a consequence, enhances VEGF-A-mediated permeability. PMID- 15345508 TI - Statin inhibition of Fc receptor-mediated phagocytosis by macrophages is modulated by cell activation and cholesterol. AB - OBJECTIVES: An inflammatory response to altered lipoproteins that accumulate in the arterial wall is a major component of the pathogenesis of atherosclerosis. Statins reduce plasma levels of low-density lipoprotein (LDL) and are effective treatments for atherosclerosis. It is hypothesized that they also modulate inflammation. The aim of this study was to examine whether lovastatin inhibits macrophage inflammatory processes and clarify its mechanism of action. METHODS AND RESULTS: We examined the effects of statins on phagocytosis of antibody coated red blood cells by cultured human monocytes and mouse peritoneal macrophages. Lovastatin, simvastatin, and zaragozic acid, a squalene synthase inhibitor, blocked Fc receptor-mediated phagocytosis by cultured human monocytes and mouse peritoneal macrophages. The inhibitory effect of lovastatin on Fc receptor-mediated phagocytosis was prevented completely by addition of mevalonate, farnesyl pyrophosphate, LDL, or cholesterol to the culture medium. The inhibitory effect of zaragozic acid was reversed by addition of LDL, but not by the addition of geranylgeranyl pyrophosphate, to the medium. In addition, the effect of lovastatin on phagocytosis is a function of cell activation because treatment of cells with tumor necrosis factor-alpha or lipopolysaccharide prevented inhibition of phagocytosis by lovastatin. CONCLUSIONS: The inhibition of Fc receptor-mediated phagocytosis of lovastatin is related to its effect on cholesterol biosynthesis rather than its effect on the formation of isoprenoids. PMID- 15345509 TI - Arterial permeability and efflux of apolipoprotein B-containing lipoproteins assessed by in situ perfusion and three-dimensional quantitative confocal microscopy. AB - OBJECTIVES: There is accumulating evidence that an increased risk of cardiovascular disease (CVD) is not simply caused by the degree of arterial exposure to plasma lipoproteins but, in addition, is determined by the affinity of the vasculature for different lipoprotein phenotypes. In this study we compare the delivery and efflux of 2 atherogenic lipoproteins to further understand the factors that regulate cholesterol accumulation in early atherogenesis. METHODS AND RESULTS: Lipoproteins containing apolipoprotein (apo) B100 (a low-density lipoprotein [LDL]) and apoB48 (chylomicron remnants) were isolated and differentially conjugated with fluorophores and simultaneously perfused at equivalent concentrations in situ through rabbit carotid vessels. Perfusion systems were established to quantify and differentiate between lipoprotein arterial delivery and efflux. The total average rate of delivery for LDL particles (23 nm) compared with chylomicron remnants (50 nm) was 4427 particles/min(-1) per microm3 and 452 particles/min(-1) per microm3, respectively. In contrast, the average rate of efflux was 3195 particles/min(-1) per microm3 and 163 particles/min(-1) per microm3 for LDL and chylomicron remnants, respectively. CONCLUSIONS: Results indicate that although LDL particles have a higher rate of delivery, they efflux more readily from arterial tissue compared with the larger chylomicron remnants. Collectively, our findings highlight that lipoproteins permeate through arterial tissue differently and may be dependent on the phenotype and potential interactions with extracellular matrix components. PMID- 15345510 TI - Id family of transcription factors and vascular lesion formation. AB - Vascular smooth muscle cell (VSMC) modulation to a de-differentiated phenotype and proliferation are key components of vascular lesion formation. Understanding how these processes are regulated is essential to understanding the progression of vascular diseases such as atherosclerosis and in-stent restenosis. The Id family of helix-loop-helix (HLH) transcription factors has emerged as important regulators of cellular growth and differentiation. Recent published findings have implicated the Id proteins as important regulators of growth and phenotypic modulation in VSMC and in the vascular response to injury. In this review, we summarize what is known regarding how the Id proteins function to control cellular growth and differentiation and their role in vascular lesion formation. PMID- 15345511 TI - Transcription factor CHF1/Hey2 regulates neointimal formation in vivo and vascular smooth muscle proliferation and migration in vitro. AB - OBJECTIVE: To determine the role of the cardiovascular-restricted, hairy-related bHLH transcription factor, CHF1/Hey2, in the biological response to vascular injury. METHODS AND RESULTS: We investigated the response of CHF1/Hey2-deficient mice to vascular injury in vivo and the response of primary cultured vascular smooth muscle cells (VSMCs) from these mice to growth factors in vitro. Neointima formation after arterial wire injury is decreased in knockout (KO) compared with wild-type (WT) mice (0.025+/-0.011 mm2 in WT [n=13]) versus 0.016+/-0.008 mm2 in KO (n=12; P<0.05) and is accompanied by reduced cellular proliferation. CHF1/Hey2 deficient VSMCs proliferate slowly compared with WT VSMCs and also show decreased migration in response to platelet-derived growth factor (PDGF) (62.6+/-10.3 CPF versus 37.2+/-13.5 CPF; P<0.01) and heparin-binding epidermal growth factor-like growth factor (HB-EGF) (27.4+/-7.7 CPF versus 6.4+/-3.7 CPF, P<0.05). Furthermore, lamellipodia formation and membrane ruffling induced by these chemoattractants are diminished in KO VSMCs, which is correlated with decreased activation of the small GTPase Rac1. Although total Rac1 protein was not changed in KO VSMCs, the level of the Rac guanine exchange factor (GEF), Sos1, was decreased. CONCLUSIONS: CHF1/Hey2 is an important regulator of vascular smooth muscle cell (VSMC) accumulation during vascular remodeling and responsiveness to growth factors in vitro. PMID- 15345512 TI - Lipoprotein(a): an elusive cardiovascular risk factor. AB - Lipoprotein (a) [Lp(a)], is present only in humans, Old World nonhuman primates, and the European hedgehog. Lp(a) has many properties in common with low-density lipoprotein (LDL) but contains a unique protein, apo(a), which is structurally different from other apolipoproteins. The size of the apo(a) gene is highly variable, resulting in the protein molecular weight ranging from 300 to 800 kDa; this large variation may be caused by neutral evolution in the absence of any selection advantage. Apo(a) influences to a major extent metabolic and physicochemical properties of Lp(a), and the size polymorphism of the apo(a) gene contributes to the pronounced heterogeneity of Lp(a). There is an inverse relationship between apo(a) size and Lp(a) levels; however, this pattern is complex. For a given apo(a) size, there is a considerable variation in Lp(a) levels across individuals, underscoring the importance to assess allele-specific Lp(a) levels. Further, Lp(a) levels differ between populations, and blacks have generally higher levels than Asians and whites, adjusting for apo(a) sizes. In addition to the apo(a) size polymorphism, an upstream pentanucleotide repeat (TTTTA(n)) affects Lp(a) levels. Several meta-analyses have provided support for an association between Lp(a) and coronary artery disease, and the levels of Lp(a) carried in particles with smaller size apo(a) isoforms are associated with cardiovascular disease or with preclinical vascular changes. Further, there is an interaction between Lp(a) and other risk factors for cardiovascular disease. The physiological role of Lp(a) is unknown, although a majority of studies implicate Lp(a) as a risk factor. PMID- 15345513 TI - Nitric oxide-releasing aspirin derivative, NCX 4016, promotes reparative angiogenesis and prevents apoptosis and oxidative stress in a mouse model of peripheral ischemia. AB - BACKGROUND: Recently, nitric oxide (NO) donors have been developed that mimic the physiological intracellular release of NO. We evaluated whether one of these new compounds, consisting of aspirin coupled to an NO-releasing moiety (NCX 4016), would protect limbs from supervening arterial occlusion. METHODS AND RESULTS: Mice were assigned to receive regular chow or chow containing NCX 4016 or aspirin (both at 300 mumol/kg body weight, daily) throughout the 3-week experimental period. One week after randomization, they underwent surgical excision of the left femoral artery. Limb blood flow recovery (laser Doppler flowmetry) was accelerated by NCX 4016 as compared with aspirin or vehicle (P<0.05). In controls, histological analysis revealed a 35% increase in the capillary density of ischemic muscles compared with contralateral ones, indicative of spontaneous angiogenesis. Neovascularization was enhanced by NCX 4016 (91%; P<0.05 versus vehicle), but not by aspirin (51%; P=NS versus vehicle). Furthermore, NCX 4016 reduced endothelial cell (EC) apoptosis (4.3+/-1.0 versus 8.7+/-2.0 in aspirin and 12.6+/-3.3 ECs/1000 cap in vehicle; P<0.05 for either comparison) as well as caspase-3 mRNA levels in ischemic muscles ([caspase-3/GAPDH]*100 = 0.09+/-0.04 versus 2.30+/-0.44 in aspirin and 2.30+/-0.32 in vehicle; P<0.01 for either comparison). Nitrite levels and the ratio of reduced to oxidized glutathione were selectively increased in ischemic muscles by NCX 4016. Vascular endothelial growth factor-A expression was reduced by aspirin, with this effect being blunted by NCX 4016. CONCLUSIONS: Pretreatment with the new oral NO-releasing aspirin derivative stimulates reparative angiogenesis and prevents apoptosis and oxidative stress, thereby alleviating the consequences of supervening arterial occlusion. PMID- 15345514 TI - Altered PDGF-BB-induced p38 MAP kinase activation in diabetic vascular smooth muscle cells: roles of protein kinase C-delta. AB - OBJECTIVE: We investigated the regulation of p38 mitogen-activated protein kinase (MAPK) by platelet-derived growth factor (PDGF)-BB and its biological effects in cultured normal and diabetic rat vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: VSMC growth from diabetic rats was faster than that from normal rats. The expression of the PDGF beta-receptor in diabetic VSMCs was significantly elevated compared with that in normal cells, and PDGF-BB-induced p38 phosphorylation in diabetic cells was more enhanced via MAPK kinase (MKK) 3/6. The level of PKC activity in diabetic cells increased more than that in normal cells with or without PDGF-BB. Although protein kinase C (PKC)-betaII and PKC-delta were activated by diabetes, PDGF-BB could further enhance the level of PKC-delta alone. PDGF-BB-induced cell migration was more elevated in diabetic VSMCs, and the increase was significantly inhibited by SB-203580, rottlerin, and antisense oligodeoxynucleotides for PKC-delta. PDGF-BB-induced p38 phosphorylation also regulated cell growth, cyclooxygenase-2 levels, and arachidonic acid release, but not apoptosis. These levels were more elevated in diabetic cells, which were inhibited by SB-203580. CONCLUSIONS: Our study established that PDGF-BB phosphorylated p38 via PKC-delta and the subsequent MKK 3/6, leading to cell growth regulation and the progression of a chronic inflammatory process in diabetic VSMCs. PMID- 15345515 TI - Enzymatic modification of low-density lipoprotein in the arterial wall: a new role for plasmin and matrix metalloproteinases in atherogenesis. AB - OBJECTIVE: Functionally interactive proteases of the plasminogen/plasmin and the matrix metalloproteinase (MMP) system degrade and reorganize the extracellular matrix of the vessel wall in atherosclerosis. Here we investigated whether such proteases are able to confer atherogenic properties onto low density lipoprotein by nonoxidative modification. METHODS AND RESULTS: Similar to the recently described enzymatically-modified low-density lipoprotein (E-LDL), native LDL exposed to plasmin or matrix MMP-2 or MMP-9 and cholesterylester-hydrolase (CEH) showed extensive deesterification, with ratios of free cholesterol to total cholesterol rising to 0.8 compared with 0.2 in native LDL. When the ratio exceeded 0.6, both plasmin/CEH-LDL and MMP/CEH-LDL fused into larger particles. In parallel, they gained C-reactive protein-dependent complement-activating capacity. E-LDL produced with any protease/CEH combination was efficiently taken up by human macrophages, whereby marked induction of MMP-2 expression by E-LDL was observed. These in vitro findings had their in vivo correlates: urokinase type plasminogen activator, MMP-2, and MMP-9 were detectable in both early and advanced human atherosclerotic lesions in colocalization with E-LDL. CONCLUSIONS: Plasmin and MMP-2/MMP-9 may not only be involved in remodeling of the extracellular matrix in progressing plaques, but they may also be involved in lipoprotein modification during genesis and progression of atherosclerotic lesions. PMID- 15345516 TI - Inhibition of tumor necrosis factor-alpha reduces atherosclerosis in apolipoprotein E knockout mice. AB - OBJECTIVE: Inflammation plays an important role in atherosclerosis. One of the most potent pro-inflammatory cytokines is tumor necrosis factor-alpha (TNF alpha), a cytokine identified to have a pathogenic role in chronic inflammatory diseases such as rheumatoid arthritis (RA). The aim of the study was to evaluate the importance of TNF-alpha in atherogenesis. METHODS AND RESULTS: Mice deficient in both apolipoprotein E (apoE) and TNF-alpha were compared regarding their atherosclerotic burden. Mice were fed a Western-style diet (WD) or normal chow. Mice deficient in both apoE and TNF-alpha exhibited a 50% (P=0.035) reduction of relative lesion size after 10 weeks of WD. Bone marrow transplantation of apoE(o) mice with apoE(o)tnf-alpha(o) bone marrow resulted in a 83% (P=0.021) reduction after 25 weeks on WD. In apoE knockout mice treated with recombinant soluble TNF receptor I releasing pellets, there was a reduction in relative lesion size after 25 weeks of 75% (P=0.018). CONCLUSIONS: These findings demonstrate that TNF-alpha is actively involved in the progression of atherosclerosis. Accordingly, TNF alpha represents a possible target for prevention of atherosclerosis. This may be of particular importance in rheumatoid arthritis because these patients have an increased risk for cardiovascular disease. PMID- 15345517 TI - Myocardin--not quite MyoD. PMID- 15345518 TI - Unique pathway for cholesterol uptake in fat cells. PMID- 15345519 TI - How vascular NAD(P)H oxidase activity and nox isoform expression are regulated. PMID- 15345520 TI - CD40L-positive platelets induce CD40L expression de novo in endothelial cells: adding a loop to microvascular inflammation. PMID- 15345521 TI - Subantimicrobial doses of tetracycline. PMID- 15345522 TI - A deeper look into single-secretory vesicle dynamics. PMID- 15345523 TI - A signal transduction pathway model prototype I: From agonist to cellular endpoint. AB - The postgenomic era is providing a wealth of information about the genes involved in many cellular processes. However, the ability to apply this information to understanding cellular signal transduction is limited by the lack of tools that quantitatively describe cellular signaling processes. The objective of the current studies is to provide a framework for modeling cellular signaling processes beginning at a plasma membrane receptor and ending with a measurable endpoint in the signaling process. Agonist-induced Ca(2+) mobilization coupled to down stream phosphorylation events was modeled using knowledge of in vitro and in vivo process parameters. The simulation process includes several modules that describe cellular processes involving receptor activation phosphoinositide metabolism, Ca(2+)-release, and activation of a calmodulin-dependent protein kinase. A Virtual Cell-based simulation was formulated using available literature data and compared to new and existing experimental results. The model provides a new approach to facilitate hypothesis-driven investigation and experimental design based upon simulation results. These investigations may be directed at the timing of multiple phosphorylation/dephosphorylation events affecting key enzymatic activities in the signaling pathway. PMID- 15345524 TI - A signal transduction pathway model prototype II: Application to Ca2+-calmodulin signaling and myosin light chain phosphorylation. AB - An agonist-initiated Ca(2+) signaling model for calmodulin (CaM) coupled to the phosphorylation of myosin light chains was created using a computer-assisted simulation environment. Calmodulin buffering was introduced as a module for directing sequestered CaM to myosin light chain kinase (MLCK) through Ca(2+) dependent release from a buffering protein. Using differing simulation conditions, it was discovered that CaM buffering allowed transient production of more Ca(2+)-CaM-MLCK complex, resulting in elevated myosin light chain phosphorylation compared to nonbuffered control. Second messenger signaling also impacts myosin light chain phosphorylation through the regulation of myosin light chain phosphatase (MLCP). A model for MLCP regulation via its regulatory MYPT1 subunit and interaction of the CPI-17 inhibitor protein was assembled that incorporated several protein kinase subsystems including Rho-kinase, protein kinase C (PKC), and constitutive MYPT1 phosphorylation activities. The effects of the different routes of MLCP regulation depend upon the relative concentrations of MLCP compared to CPI-17, and the specific activities of protein kinases such as Rho and PKC. Phosphorylated CPI-17 (CPI-17P) was found to dynamically control activity during agonist stimulation, with the assumption that inhibition by CPI 17P (resulting from PKC activation) is faster than agonist-induced phosphorylation of MYPT1. Simulation results are in accord with literature measurements of MLCP and CPI-17 phosphorylation states during agonist stimulation, validating the predictive capabilities of the system. PMID- 15345525 TI - Probing protein mechanics: residue-level properties and their use in defining domains. AB - It is becoming clear that, in addition to structural properties, the mechanical properties of proteins can play an important role in their biological activity. It nevertheless remains difficult to probe these properties experimentally. Whereas single-molecule experiments give access to overall mechanical behavior, notably the impact of end-to-end stretching, it is currently impossible to directly obtain data on more local properties. We propose a theoretical method for probing the mechanical properties of protein structures at the single-amino acid level. This approach can be applied to both all-atom and simplified protein representations. The probing leads to force constants for local deformations and to deformation vectors indicating the paths of least mechanical resistance. It also reveals the mechanical coupling that exists between residues. Results obtained for a variety of proteins show that the calculated force constants vary over a wide range. An analysis of the induced deformations provides information that is distinct from that obtained with measures of atomic fluctuations and is more easily linked to residue-level properties than normal mode analyses or dynamic trajectories. It is also shown that the mechanical information obtained by residue-level probing opens a new route for defining so-called dynamical domains within protein structures. PMID- 15345526 TI - Neutron frequency windows and the protein dynamical transition. AB - Proteins undergo an apparent dynamical transition on temperature variation that has been correlated with the onset of function. The transition in the mean-square displacement, , that is observed using a spectrometer or computer simulation, depends on the relationship between the timescales of the relaxation processes activated and the timescale accessible to the instrument or simulation. Models are described of two extreme situations---an "equilibrium" model, in which the long-time dynamics changes with temperature and all motions are resolved by the instrument used; and a "frequency window" model, in which there is no change in the long-time dynamics but as the temperature increases, the relaxation frequencies move into the instrumental range. Here we demonstrate that the latter, frequency-window model can describe the temperature and timescale dependences of both the intermediate neutron scattering function and derived from molecular dynamics simulations of a small protein in a cryosolution. The frequency-window model also describes the energy-resolution and temperature dependences of obtained from experimental neutron scattering on glutamate dehydrogenase in the same solvent. Although equilibrium effects should also contribute to dynamical transitions in proteins, the present results suggests that frequency-window effects can play a role in the simulations and experiments examined. Finally, misquotations of previous findings are discussed in the context of solvent activation of protein dynamics and the possible relationship of this to activity. PMID- 15345527 TI - Lipid bilayer topology of the transmembrane alpha-helix of M13 Major coat protein and bilayer polarity profile by site-directed fluorescence spectroscopy. AB - This article presents a new formalism to perform a quantitative fluorescence analysis using the Stokes shift of AEDANS-labeled cysteine mutants of M13 major coat protein incorporated in lipid bilayers. This site-directed fluorescence spectroscopy approach enables us to obtain the topology of the bilayer-embedded transmembrane alpha-helix from the orientation and tilt angles, and relative bilayer location. Both in pure dioleoylphosphatidylcholine and dioleoylphosphatidylcholine/dioleoylphosphatidylglycerol (4:1 mol/mol) bilayers, which have a similar bilayer thickness, the tilt angle of the transmembrane helix of the coat protein turns out to be 23 degrees +/- 4. Upon decreasing the hydrophobic thickness on going from dieicosenoylphosphatidylcholine to dimyristoylphosphatidylcholine, the tilt angle and orientation angle of the transmembrane alpha-helix change. The protein responds to an increase of hydrophobic stress by increasing the tilt angle so as to keep much of its hydrophobic part inside the bilayer. At the same time, the transmembrane helix rotates at its long axis so as to optimize the hydrophobic and electrostatic interactions of the C-terminal phenylalanines and lysines, respectively. The increase of tilt angle cannot completely keep the hydrophobic protein section within the bilayer, but the C-terminal part remains anchored at the acyl chain/glycerol backbone interface at the cost of the N-terminal section. In addition, our analysis results in the profile of the dielectric constant of the hydrophobic domain of the bilayer. For all phospholipid bilayers studied the profile has a concave shape, with a value of the dielectric constant of 4.0 in the center of the bilayer. The dielectric constant increases on approaching the headgroup region with a value of 12.4 at the acyl-chain/glycerol backbone interface for the various phosphatidylcholines with different chain lengths. For dioleoylphosphatidylcholine/dioleoylphosphatidylglycerol (4:1 mol/mol) bilayers the value of the dielectric constant at the acyl-chain/glycerol backbone interface is 18.6. In conclusion, the consistency of our analysis shows that the applied cysteine-scanning mutagenesis method with AEDANS labeling of a helical transmembrane protein in combination with a quantitative formalism offers a reliable description of the lipid bilayer topology of the protein and bilayer properties. This also indicates that the spacer link between the protein and AEDANS label is long enough to monitor the local polarity of the lipid environment and not that of the amino-acid residues of the protein, and short enough to have the topology of the protein imposing on the fluorescence properties of the AEDANS label. PMID- 15345528 TI - Molecular dynamics simulations of forced conformational transitions in 1,6-linked polysaccharides. AB - Recent atomic force microscopy stretching measurements of single polysaccharide molecules suggest that their elasticity is governed by force-induced conformational transitions of the pyranose ring. However, the mechanism of these transitions and the mechanics of the pyranose ring are not fully understood. Here we use steered molecular dynamics simulations of the stretching process to unravel the mechanism of forced conformational transitions in 1,6 linked polysaccharides. In contrast to most sugars, 1,6 linked polysaccharides have an extra bond in their inter-residue linkage, C5-C6, around which restricted rotations occur and this additional degree of freedom increases the mechanical complexity of these polymers. By comparing the computational results with the atomic force microscopy data we determine that forced rotations around the C5-C6 bond have a significant and different impact on the elasticity of alpha- and beta linked polysaccharides. Beta-linkages of a polysaccharide pustulan force the rotation around the C5-C6 bonds and produce a Hookean-like elasticity but do not affect the conformation of the pyranose rings. However, alpha-linkages of dextran induce compound conformational transitions that include simultaneous rotations around the C5-C6 bonds and chair-boat transitions of the pyranose rings. These previously not-recognized transitions are responsible for the characteristic plateau in the force-extension relationship of dextran. PMID- 15345529 TI - Physics of RecA-mediated homologous recognition. AB - To accomplish its DNA strand exchange activities, the Escherichia coli protein RecA polymerizes onto DNA to form a stiff helical nucleoprotein filament within which the DNA is extended by 50%. Homology search and recognition occurs between ssDNA within the filament and an external dsDNA molecule. We show that stretching the internal DNA greatly enhances homology recognition by increasing the probability that the homologous regions of a stretched DNA molecule and a parallel, unstretched DNA molecule will be "in register" at some position. We also show that the stretching and stiffness of the filament act together to ensure that initiation of homologous exchange between the substrate DNA molecules at one position precludes initiation of homologous exchange at any other position. This prevents formation of multiple exchange site "topological traps" which would prevent completion of the exchange reaction and resolution of the products. PMID- 15345530 TI - Dependence of DNA polymerase replication rate on external forces: a model based on molecular dynamics simulations. AB - Molecular dynamics simulations are presented for a Thermus aquaticus (Taq) DNA polymerase I complex (consisting of the protein, the primer-template DNA strands, and the incoming nucleotide) subjected to external forces. The results obtained with a force applied to the DNA template strand provide insights into the effect of the tension on the activity of the enzyme. At forces below 30 pN a local model based on the parameters determined from the simulations, including the restricted motion of the DNA bases at the active site, yields a replication rate dependence on force in agreement with experiment. Simulations above 40 pN reveal large conformational changes in the enzyme-bound DNA that may have a role in the force induced exonucleolysis observed experimentally. PMID- 15345531 TI - Michaelis-Menten kinetics under spatially constrained conditions: application to mibefradil pharmacokinetics. AB - Two different approaches were used to study the kinetics of the enzymatic reaction under heterogeneous conditions to interpret the unusual nonlinear pharmacokinetics of mibefradil. Firstly, a detailed model based on the kinetic differential equations is proposed to study the enzymatic reaction under spatial constraints and in vivo conditions. Secondly, Monte Carlo simulations of the enzyme reaction in a two-dimensional square lattice, placing special emphasis on the input and output of the substrate were applied to mimic in vivo conditions. Both the mathematical model and the Monte Carlo simulations for the enzymatic reaction reproduced the classical Michaelis-Menten (MM) kinetics in homogeneous media and unusual kinetics in fractal media. Based on these findings, a time dependent version of the classic MM equation was developed for the rate of change of the substrate concentration in disordered media and was successfully used to describe the experimental plasma concentration-time data of mibefradil and derive estimates for the model parameters. The unusual nonlinear pharmacokinetics of mibefradil originates from the heterogeneous conditions in the reaction space of the enzymatic reaction. The modified MM equation can describe the pharmacokinetics of mibefradil as it is able to capture the heterogeneity of the enzymatic reaction in disordered media. PMID- 15345532 TI - A computational model of the human left-ventricular epicardial myocyte. AB - A computational model of the human left-ventricular epicardial myocyte is presented. Models of each of the major ionic currents present in these cells are formulated and validated using experimental data obtained from studies of recombinant human ion channels and/or whole-cell recording from single myocytes isolated from human left-ventricular subepicardium. Continuous-time Markov chain models for the gating of the fast Na(+) current, transient outward current, rapid component of the delayed rectifier current, and the L-type calcium current are modified to represent human data at physiological temperature. A new model for the gating of the slow component of the delayed rectifier current is formulated and validated against experimental data. Properties of calcium handling and exchanger currents are altered to appropriately represent the dynamics of intracellular ion concentrations. The model is able to both reproduce and predict a wide range of behaviors observed experimentally including action potential morphology, ionic currents, intracellular calcium transients, frequency dependence of action-potential duration, Ca(2+)-frequency relations, and extrasystolic restitution/post-extrasystolic potentiation. The model therefore serves as a useful tool for investigating mechanisms of arrhythmia and consequences of drug-channel interactions in the human left-ventricular myocyte. PMID- 15345533 TI - In silico activation of KcsA K+ channel by lateral forces applied to the C termini of inner helices. AB - Crystallographic studies of K(+) channels in the closed (KcsA) and open (MthK) states suggest that Gly(99) (KcsA numbering) in the inner helices serves as a gating hinge during channel activation. However, some P-loop channels have larger residues in the corresponding position. The comparison of x-ray structures of KcsA and MthK shows that channel activation alters backbone torsions and helical H-bonds in residues 95-105. Importantly, the changes in Gly(99) are not the largest ones. This raises questions about the mechanism of conformational changes upon channel gating. In this work, we have built a model of the open KcsA using MthK as a template and simulated opening and closing of KcsA by constraining C ends of the inner helices at a gradually changing distance from the pore axis without restraining mobility of the helices along the axis. At each imposed distance, the energy was Monte Carlo-minimized. The channel-opening and channel closing trajectories arrived to the structures in which the backbone geometry was close to that seen in MthK and KcsA, respectively. In the channel-opening trajectory, the constraints-induced lateral forces caused kinks at midpoints of the inner helices between Val(97) and Gly(104) but did not destroy interdomain contacts, the pore helices, and the selectivity filter. The simulated activation of the Gly(99)Ala mutant yielded essentially similar results. Analysis of interresidue energies shows that the N-terminal parts of the inner helices form strong attractive contacts with the pore helices and the outer helices. The lateral forces induce kinks at the position where the helix-breaking torque is maximal and the intersegment contacts vanish. This mechanism may be conserved in different P-loop channels. PMID- 15345534 TI - Coupling of protein relaxation to ligand binding and migration in myoglobin. AB - Protein relaxation, ligand binding, and ligand migration into a hydrophobic cavity in myoglobin are unified by a bounded diffusion model which produces an accurate fit to complex ligand rebinding data over eight decades in time and a 160 K temperature range, in qualitative agreement with time-resolved x-ray crystallography. Protein relaxation operates in a cyclic manner to move the ligand away from the binding site. PMID- 15345535 TI - Influence of the solvent structure on the electrostatic interactions in proteins. AB - The proper estimation of the influence of the many-body dynamic solvent microstructure on a pairwise electrostatic interaction (PEI) at the protein solvent interface is very important for solving many biophysical problems. In this work, the PEI energy was calculated for a system that models the interface between a protein and an aqueous solvent. The concept of nonlocal electrostatics for interfacial electrochemical systems was used to evaluate the contribution of a solvent orientational polarization, correlated by the network of hydrogen bonds, into the PEI energy in proteins. The analytical expression for this energy was obtained in the form of Coulomb's law with an effective distance-dependent dielectric function. The asymptotic and numerical analysis carried out for this function revealed several features of dielectric heterogeneity at the protein solvent interface. For charges located in close proximity to this interface, the values of the dielectric function for the short-distance electrostatic interactions were found to be remarkably smaller than those determined by the classical model, in which the solvent was considered as the uniform dielectric medium of high dielectric constant. Our results have shown that taking into consideration the dynamic solvent microstructure remarkably increases the value of the PEI energy at the protein-solvent interface. PMID- 15345536 TI - Continuum diffusion reaction rate calculations of wild-type and mutant mouse acetylcholinesterase: adaptive finite element analysis. AB - As described previously, continuum models, such as the Smoluchowski equation, offer a scalable framework for studying diffusion in biomolecular systems. This work presents new developments in the efficient solution of the continuum diffusion equation. Specifically, we present methods for adaptively refining finite element solutions of the Smoluchowski equation based on a posteriori error estimates. We also describe new, molecular-surface-based models, for diffusional reaction boundary criteria and compare results obtained from these models with the traditional spherical criteria. The new methods are validated by comparison of the calculated reaction rates with experimental values for wild-type and mutant forms of mouse acetylcholinesterase. The results show good agreement with experiment and help to define optimal reactive boundary conditions. PMID- 15345537 TI - LMProt: an efficient algorithm for Monte Carlo sampling of protein conformational space. AB - A new and efficient Monte Carlo algorithm for sampling protein configurations in the continuous space is presented; the efficiency of this algorithm, named Local Moves for Proteins (LMProt), was compared to other alternative algorithms. For this purpose, we used an intrachain interaction energy function that is proportional to the root mean square deviation (rmsd) with respect to alpha carbons from native structures of real proteins. For phantom chains, the LMProt method is approximately 10(4) and 20 times faster than the algorithms Thrashing (no local moves) and Sevenfold Way (local moves), respectively. Additionally, the LMProt was tested for real chains (excluded-volume all-atoms model); proteins 5NLL (138 residues) and 1BFF (129 residues) were used to determine the folding success xi as a function of the number eta of residues involved in the chain movements, and as a function of the maximum amplitude of atomic displacement delta r(max). Our results indicate that multiple local moves associated with relative chain flexibility, controlled by appropriate adjustments for eta and delta r(max), are essential for configurational search efficiency. PMID- 15345538 TI - Effects of receptor interaction in bacterial chemotaxis. AB - Signaling in bacterial chemotaxis is mediated by several types of transmembrane chemoreceptors. The chemoreceptors form tight polar clusters whose functions are of great biological interest. Here, we study the general properties of a chemotaxis model that includes interaction between neighboring chemoreceptors within a receptor cluster and the appropriate receptor methylation and demethylation dynamics to maintain (near) perfect adaptation. We find that, depending on the receptor coupling strength, there are two steady-state phases in the model: a stationary phase and an oscillatory phase. The mechanism for the existence of the two phases is understood analytically. Two important phenomena in transient response, the overshoot in response to a pulse stimulus and the high gain in response to sustained changes in external ligand concentrations, can be explained in our model, and the mechanisms for these two seemingly different phenomena are found to be closely related. The model also naturally accounts for several key in vitro response experiments and the recent in vivo fluorescence resonance energy transfer experiments for various mutant strains. Quantitatively, our study reveals possible choices of parameters for fitting the existing experiments and suggests future experiments to test the model predictions. PMID- 15345539 TI - Simulating induced interdigitation in membranes. AB - In this study we introduce a mesoscopic lipid-water-alcohol model. Dissipative particle dynamics (DPD) simulations have been used to investigate the induced interdigitation of bilayers consisting of double-tail lipids by adding alcohol molecules to the bilayer. Our simulations nicely reproduce the experimental phase diagrams. We find that alcohol can induce an interdigitated structure where the common bilayer structure changes into monolayer in which the alcohol molecules screen the hydrophobic tails from the water phase. At low concentrations of alcohol the membrane has domains of the interdigitated phase that are in coexistence with the common membrane phase. We compute the effect of the chain length of the alcohol on the phase behavior of the membrane and show that the stability of the interdigitated phase depends on the length of the alcohol. We show that we can reproduce the experimental hydrophobic thickness of the bilayer for various combinations of lipids and alcohols. We use our model to clarify some of the experimental questions related to the structure of the interdigitated phase and put forward a simple model that explains the alcohol chain length dependence of the stability of this interdigitated phase. PMID- 15345540 TI - A model of effective diffusion and tortuosity in the extracellular space of the brain. AB - Tortuosity of the extracellular space describes hindrance posed to the diffusion process by a geometrically complex medium in comparison to an environment free of any obstacles. Calculating tortuosity in biologically relevant geometries is difficult. Yet this parameter has proved very important for many processes in the brain, ranging from ischemia and osmotic stress to delivery of nutrients and drugs. It is also significant for interpretation of the diffusion-weighted magnetic resonance data. We use a volume-averaging procedure to obtain a general expression for tortuosity in a complex environment. A simple approximation then leads to tortuosity estimates in a number of two-dimensional (2D) and three dimensional (3D) geometries characterized by narrow pathways between the cellular elements. It also explains the counterintuitive fact of lower diffusion hindrance in a 3D environment. Comparison with Monte Carlo numerical simulations shows that the model gives reasonable tortuosity estimates for a number of regular and randomized 2D and 3D geometries. Importantly, it is shown that addition of dead end pores increases tortuosity in proportion to the square root of enlarged total extracellular volume fraction. This conclusion is further supported by the previously described tortuosity decrease in ischemic brain slices where dead-end pores were partially occluded by large macromolecules introduced into the extracellular space. PMID- 15345541 TI - How optimal are the binding energetics of barnase and barstar? AB - The extracellular ribonuclease barnase and its intracellular inhibitor barstar bind fast and with high affinity. Although extensive experimental and theoretical studies have been carried out on this system, it is unclear what the relative importance of different contributions to the high affinity is and whether binding can be improved through point mutations. In this work, we first applied Poisson Boltzmann electrostatic calculations to 65 barnase-barstar complexes with mutations in both barnase and barstar. The continuum electrostatic calculations with a van der Waals surface dielectric boundary definition result in the electrostatic interaction free energy providing the dominant contribution favoring barnase-barstar binding. The results show that the computed electrostatic binding free energy can be improved through mutations at W44/barstar and E73/barnase. Furthermore, the determinants of binding affinity were quantified by applying COMparative BINding Energy (COMBINE) analysis to derive quantitative structure-activity relationships (QSARs) for the 65 complexes. The COMBINE QSAR model highlights approximately 20 interfacial residue pairs as responsible for most of the differences in binding affinity between the mutant complexes, mainly due to electrostatic interactions. Based on the COMBINE model, together with Brownian dynamics simulations to compute diffusional association rate constants, several mutants were designed to have higher binding affinities than the wild-type proteins. PMID- 15345542 TI - Rational design of solution additives for the prevention of protein aggregation. AB - We have developed a statistical-mechanical model of the effect of solution additives on protein association reactions. This model incorporates solvent radial distribution functions obtained from all-atom molecular dynamics simulations of particular proteins into simple models of protein interactions. In this way, the effects of additives can be computed along the entire association/dissociation reaction coordinate. We used the model to test our hypothesis that a class of large solution additives, which we term "neutral crowders," can slow protein association and dissociation by being preferentially excluded from protein-protein encounter complexes, in a manner analogous to osmotic stress. The magnitude of this proposed "gap effect" was probed for two simple model systems: the association of two spheres and the association of two planes. Our results suggest that for a protein of 20 A radius, an 8 A additive can increase the free energy barrier for association and dissociation by as much as 3-6 kcal/mol. Because the proposed gap effect is present only for reactions involving multiple molecules, it can be exploited to develop novel additives that affect protein association reactions although having little or no effect on unimolecular reactions such as protein folding. This idea has many potential applications in areas such as the stabilization of proteins against aggregation during folding and in pharmaceutical formulations. PMID- 15345543 TI - Kinetics of target site localization of a protein on DNA: a stochastic approach. AB - It is widely recognized that the cleaving rate of a restriction enzyme on target DNA sequences is several orders-of-magnitude faster than the maximal one calculated from the diffusion-limited theory. It was therefore commonly assumed that the target site interaction of a restriction enzyme with DNA has to occur via two steps: one-dimensional diffusion along a DNA segment, and long-range jumps coming from association-dissociation events. We propose here a stochastic model for this reaction which comprises a series of one-dimensional diffusions of a restriction enzyme on nonspecific DNA sequences interrupted by three dimensional excursions in the solution until the target sequence is reached. This model provides an optimal finding strategy which explains the fast association rate. Modeling the excursions by uncorrelated random jumps, we recover the expression of the mean time required for target site association to occur given by Berg et al. in 1981, and we explicitly give several physical quantities describing the stochastic pathway of the enzyme. For competitive target sites we calculate two quantities: processivity and preference. By comparing these theoretical expressions to recent experimental data obtained for EcoRV-DNA interaction, we quantify: 1), the mean residence time per binding event of EcoRV on DNA for a representative one-dimensional diffusion coefficient; 2), the average lengths of DNA scanned during the one-dimensional diffusion (during one binding event and during the overall process); and 3), the mean time and the mean number of visits needed to go from one target site to the other. Further, we evaluate the dynamics of DNA cleavage with regard to the probability for the restriction enzyme to perform another one-dimensional diffusion on the same DNA substrate following a three-dimensional excursion. PMID- 15345544 TI - Direct simulation of transmembrane helix association: role of asparagines. AB - The forces contributing to the association of transmembrane helices in folded membrane proteins have received considerable attention recently. In this study we investigate the importance of hydrogen bonding by studying the effect of a single Asn residue in the center of an otherwise hydrophobic transmembrane peptide using computer simulations. We use the model peptide MS1 which has been derived from the leucine zipper coiled-coil dimer of the transcription factor peptide GCN4-P1. We follow the trajectory of 36 initially monomeric MS1 transmembrane helical peptides in a membrane-mimicking octane layer as they associate into larger structures. These peptides predominately form dimers. The interaction between the polar asparagine residues, capable of simultaneously being a hydrogen-bond donor and acceptor, contributes strongly to the stability of associated helices. Only dimers with interhelical hydrogen bonds form stable structures, whereas aggregates without any hydrogen-bonding interactions form very transient structures. We examine the hydrogen-bonding patterns and find that there are two forms of dimer, one with symmetric hydrogen bonds and one with asymmetric hydrogen bonds. Based on the structures in our simulation we propose a model with a monomer <--> symmetric dimer <--> asymmetric dimer <--> trimer equilibrium. PMID- 15345545 TI - Model-based fitting of single-channel dwell-time distributions. AB - Single-channel recordings provide unprecedented resolutions on kinetics of conformational changes of ion channels. Several approaches exist for analysis of the data, including the dwell-time histogram fittings and the full maximal likelihood approaches that fit either the idealized dwell-time sequence or more ambitiously the noisy data directly using hidden Markov modeling. Although the full maximum likelihood approaches are statistically advantageous, they can be time-consuming especially for large datasets and/or complex models. We present here an alternative approach for model-based fitting of one-dimensional and two dimensional dwell-time histograms. To improve performance, we derived analytical expressions for the derivatives of one-dimensional and two-dimensional dwell-time distribution functions and employed the gradient-based variable metric method for fast search of optimal rate constants in a model. The algorithm also has the ability to allow for a first-order correction for the effects of missed events, global fitting across different experimental conditions, and imposition of typical constraints on rate constants including microscopic reversibility. Numerical examples are presented to illustrate the performance of the algorithm, and comparisons with the full maximum likelihood fitting are discussed. PMID- 15345546 TI - Wide nanoscopic pore of maxi-anion channel suits its function as an ATP conductive pathway. AB - The newly proposed function of the maxi-anion channel as a conductive pathway for ATP release requires that its pore is sufficiently large to permit passage of a bulky ATP(4-) anion. We found a linear relationship between relative permeability of organic anions of different size and their relative ionic mobility (measured as the ratio of ionic conductance) with a slope close to 1, suggesting that organic anions tested with radii up to 0.49 nm (lactobionate) move inside the channel by free diffusion. In the second approach, we, for the first time, succeeded in pore sizing by the nonelectrolyte exclusion method in single-channel patch-clamp experiments. The cutoff radii of PEG molecules that could access the channel from intracellular (1.16 nm) and extracellular (1.42 nm) sides indicated an asymmetry of the two entrances to the channel pore. Measurements by symmetrical two-sided application of PEG molecules yielded an average functional pore radius of approximately 1.3 nm. These three estimates are considerably larger than the radius of ATP(4-) (0.57-0.65 nm) and MgATP(2-) (approximately 0.60 nm). We therefore conclude that the nanoscopic maxi-anion channel pore provides sufficient room to accommodate ATP and is well suited to its function as a conductive pathway for ATP release in cell-to-cell communication. PMID- 15345547 TI - Exterior site occupancy infers chloride-induced proton gating in a prokaryotic homolog of the ClC chloride channel. AB - The ClC family of anion channels mediates the efficient, selective permeation of Cl(-) across the biological membranes of living cells under the driving force of an electrochemical gradient. In some eukaryotes, these channels are known to exhibit a unique gating mechanism, which appears to be triggered by the permeant Cl(-) anion. We infer details of this gating mechanism by studying the free energetics of Cl(-) occupancy in the pore of a prokaryotic ClC homolog. These free energetics were gleaned from 30 ns of molecular dynamics simulation on an approximately 133,000-atom system consisting of a hydrated membrane embedded StClC transporter. The binding sites for Cl(-) in the transporter were determined for the cases where the putative gating residue, Glu(148), was protonated and unprotonated. When the glutamate gate is protonated, Cl(-) favorably occupies an exterior site, S(ext), to form a queue of anions in the pore. However, when the glutamate gate is unprotonated, Cl(-) cannot occupy this site nor, consequently, pass through the pore. An additional, previously undetected, site was found in the pore near the outer membrane that exists regardless of the protonation state of Glu(148). Although this suggests that, for the prokaryotic homolog, protonation of Glu(148) may be the first step in transporting Cl(-) at the expense of H(+) transport in the opposite direction, an evolutionary argument might suggest that Cl(-) opens the ClC gate in eukaryotic channels by inducing the conserved glutamate's protonation. During an additional 20 ns free dynamics simulation, the newly discovered outermost site, S(out), and the innermost site, S(int), were seen to allow spontaneous exchange of Cl(-) ions with the bulk electrolyte while under depolarization conditions. PMID- 15345548 TI - Proton conductance of influenza virus M2 protein in planar lipid bilayers. AB - Purified M2 protein from the Udorn strain of influenza virus was reconstituted into planar lipid bilayers from liposomes. In 1 mM HCl, the single-channel conductance was measured as 6 pS with open probability of < or =0.03. The current voltage curve is linear over the achievable voltage range. The current amplitude is amantadine sensitive. In HCl solutions, the single-channel current was essentially invariant with changes in [Cl(-)], [Na(+)], and [tetraethylammonium] ([TEA(+)]), but dependent on [H(+)]. The reversal potential, determined with asymmetrical hydrogen chloride solution, is very close to the equilibrium potential of hydrogen. This appears to be the first report of single-channel proton currents with the full-length M2 protein. PMID- 15345549 TI - Conductance studies on trichotoxin_A50E and implications for channel structure. AB - Trichotoxin_A50E is an 18-residue peptaibol whose crystal structure has recently been determined. In this study, the conductance properties of trichotoxin_A50E have been investigated in neutral planar lipid bilayers. The macroscopic current voltage curves disclose a moderate voltage-sensitivity and the concentration dependence suggests the channels are primarily hexameric. Under ion gradients, shifts of the reversal potential indicate that cations are preferentially transported. Trichotoxin displays only one single-channel conductance state in a given experiment, but an ensemble of experiments reveals a distribution of conductance levels. This contrasts with the related peptaibol alamethicin, which produces multiple channel levels in a single experiment, indicative of recruitment of additional monomers into different multimeric-sized channels. Based on these conductance measurements and on the recently available crystal structure of trichotoxin_A50E, which is a shorter and straighter helix than alamethicin, a tightly-packed hexameric model structure has been constructed for the trichotoxin channel. It has molecular dimensions and surface electrostatic potential compatible with the observed conductance properties of the most probable and longer-lived channel. PMID- 15345550 TI - Interaction of poly(L-lysine)-g-poly(ethylene glycol) with supported phospholipid bilayers. AB - Interactions between the graft copolymer poly(L-lysine)-g-poly(ethylene glycol), PLL-g-PEG, and two kinds of surface-supported lipidic systems (supported phospholipid bilayers and supported vesicular layers) were investigated by a combination of microscopic and spectroscopic techniques. It was found that the application of the copolymer to zwitterionic or negatively charged supported bilayers in a buffer of low ionic strength led to their decomposition, with the resulting formation of free copolymer-lipid complexes. The same copolymer had no destructive effect on a supported vesicular layer made up of vesicles of identical composition. A comparison between poly(L-lysine), which did not induce decomposition of supported bilayers, and PLL-g-PEG copolymers with various amounts of PEG side chains per backbone lysine unit, suggested that steric repulsion between the PEG chains that developed upon adsorption of the polymer to the nearly planar surface of a supported phospholipid bilayer (SPB) was one of the factors responsible for the destruction of the SPBs by the copolymer. Other factors included the ionic strength of the buffer used and the quality of the bilayers, pointing toward the important role defects present in the SPBs play in the decomposition process. PMID- 15345551 TI - The 4,5-double bond of ceramide regulates its dipole potential, elastic properties, and packing behavior. AB - The biological activities of ceramides show a large variation with small changes in molecular structure. To help understand how the structure regulates the activity of this important lipid second messenger, we investigated the interfacial features of a series of synthetic ceramide analogs in monomolecular films at the argon-buffer interface. To minimize differences arising from the N acyl moiety, each analog had either a N-hexadecanoyl or a N-cis-4-hexadecenoyl moiety amide linked to the nitrogen of the sphingosine backbone. We found that the trans 4,5-unsaturation in the sphingosine backbone promoted closer packing and lower compressibilities of ceramide analogs in interfaces relative to comparable saturated species. Moreover, structures with this feature exhibited dipole potentials as much as 150-250 mV higher than comparable compounds lacking 4,5-unsaturation. The results support the hypothesis by M.C. Yappert and co workers that trans unsaturation in the vicinity of C4 of the sphingoid backbone augments intramolecular hydration/hydrogen bonding in the polar region. This intramolecular hydration may allow the close packing of the ceramide molecules and engender their high dipole potentials. These properties of ceramides and their analogs may be important determinants of biological function. PMID- 15345552 TI - Insertion of Alzheimer's A beta 40 peptide into lipid monolayers. AB - The amyloid beta (A beta) peptide is the major component found in the amyloid deposits in the brains of Alzheimer's disease patients. In vitro studies have demonstrated that the aggregation of A beta can take place at three orders of magnitude lower concentrations in the presence of phospholipid molecules compared to bulk peptide studies, suggesting that membrane lipids may mediate A beta toxicity. To understand the interaction of A beta with lipid membranes, we have examined A beta 40 with anionic dipalmitoylphosphatidylglycerol (DPPG), zwitterionic dipalmitoylphosphatidylcholine (DPPC), and cationic dipalmitoyltrimethylammonium propane (DPTAP) monolayers under different subphase conditions. We have used a constant surface pressure insertion assay to assess the degree of peptide insertion into the lipids. Simultaneously, we monitored the surface morphology of the monolayers with fluorescence microscopy. We have also performed dual-probe fluorescence measurements where both the peptide and lipid are tagged with chromophores. Isotherm measurements show that A beta inserts into both DPTAP and DPPG monolayers under physiologically relevant conditions. Insertion into DPPC occurs at lipid densities below that found in a bilayer. The level of insertion is inversely proportional to the lipid packing density. Our results indicate that lipids need not be anionic to interact with A beta. Electrostatic effects involved in A beta 40-lipid interaction are discussed. PMID- 15345553 TI - Domains in binary SOPC/POPE lipid mixtures studied by pulsed field gradient 1H MAS NMR. AB - We studied domain formation in mixtures of the monounsaturated lipids SOPC and POPE as a function of temperature and composition by NMR. Magic angle spinning at kHz frequencies restored resolution of (1)H NMR lipid resonances in the fluid phase, whereas the linewidth of gel-phase lipids remained rather broad and spinning frequency dependent. In regions of fluid- and gel-phase coexistence, spectra are a superposition of resonances from fluid and gel domains, as indicated by the existence of isosbestic points. Quantitative determination of the amount of lipid in the coexisting phases is straightforward and permitted construction of a binary phase diagram. Lateral rates of lipid diffusion were determined by (1)H MAS NMR with pulsed field gradients. At the onset of the phase transition near 25 degrees C apparent diffusion rates became diffusion time dependent, indicating that lipid movement is obstructed by the formation of gel phase domains. A percolation threshold at which diffusion of fluid-phase lipid becomes confined to micrometer-size domains was observed when approximately 40% of total lipid had entered the gel phase. The results indicate that common phosphatidylethanolamines may trigger domain formation in membranes within a physiologically relevant temperature range. This novel NMR approach may aid the study of lipid rafts. PMID- 15345554 TI - Oleic and docosahexaenoic acid differentially phase separate from lipid raft molecules: a comparative NMR, DSC, AFM, and detergent extraction study. AB - We have previously suggested that the omega-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA) may in part function by enhancing membrane lipid phase separation into lipid rafts. Here we further tested for differences in the molecular interactions of an oleic (OA) versus DHA-containing phospholipid with sphingomyelin (SM) and cholesterol (CHOL) utilizing (2)H NMR spectroscopy, differential scanning calorimetry, atomic force microscopy, and detergent extractions in model bilayer membranes. (2)H NMR and DSC (differential scanning calorimetry) established the phase behavior of the OA-containing 1 [(2)H(31)]palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (16:0-18:1PE d(31))/SM (1:1) and the DHA-containing 1-[(2)H(31)]palmitoyl-2-docosahexaenoyl-sn glycero-3-phosphoethanolamine (16:0-22:6PE-d(31))/SM (1:1) in the absence and presence of equimolar CHOL. CHOL was observed to affect the OA-containing phosphatidylethanolamine (PE) more than the DHA-containing PE, as exemplified by >2 x greater increase in order measured for the perdeuterated palmitic chain in 16:0-18:1PE-d(31)/SM (1:1) compared to 16:0-22:6PE-d(31)/SM (1:1) bilayers in the liquid crystalline phase. Atomic force microscopy (AFM) experiments showed less lateral phase separation between 16:0-18:1PE-rich and SM/CHOL-rich raft domains in 16:0-18:1PE/SM/CHOL (1:1:1) bilayers than was observed when 16:0-22:6PE replaced 16:0-18:1PE. Differences in the molecular interaction of 16:0-18:1PE and 16:0-22:6PE with SM/CHOL were also found using biochemical detergent extractions. In the presence of equimolar SM/CHOL, 16:0-18:1PE showed decreased solubilization in comparison to 16:0-22:6PE, indicating greater phase separation with the DHA PE. Detergent experiments were also conducted with cardiomyocytes fed radiolabeled OA or DHA. Although both OA and DHA were found to be largely detergent solubilized, the amount of OA that was found to be associated with raft rich detergent-resistant membranes exceeded DHA by almost a factor of 2. We conclude that the OA-PE phase separates from rafts far less than DHA-PE, which may have implications for cellular signaling. PMID- 15345555 TI - Nucleotide dependent intrinsic fluorescence changes of W29 and W36 in smooth muscle myosin. AB - The intrinsic fluorescence of smooth muscle myosin is sensitive to both nucleotide binding and hydrolysis. We have examined this relationship by making MDE mutants containing a single tryptophan residue at each of the seven positions found in the wild-type molecule. Previously, we have demonstrated that a conserved tryptophan residue (W512) is a major contributor to nucleotide dependent changes of intrinsic fluorescence in smooth muscle myosin. In this study, an MDE containing all the endogenous tryptophans except W512 (W512 KO-MDE) decreases in intrinsic fluorescence upon nucleotide binding, demonstrating that the intrinsic fluorescence enhancement of smooth muscle myosin is not solely due to W512. Candidates for the observed quench of intrinsic fluorescence in W512 KO MDE include W29 and W36. Whereas the intrinsic fluorescence of W36-MDE is only slightly sensitive to nucleotide binding, that of W29-MDE is paradoxically both quenched and blue-shifted upon nucleotide binding. Steady-state and time-resolved experiments suggest that fluorescence intensity changes of W29 involve both excited-state and ground-state quenching mechanisms. These results have important implications for the role of the N-terminal domain (residues 1-76) in smooth muscle myosin in the molecular mechanism of muscle contraction. PMID- 15345556 TI - Nanometer localization of single green fluorescent proteins: evidence that myosin V walks hand-over-hand via telemark configuration. AB - Myosin V is a homodimeric motor protein involved in trafficking of vesicles in the cell. It walks bipedally along actin filaments, moving cargo approximately 37 nm per step. We have measured the step size of individual myosin heads by fusing an enhanced green fluorescent protein (eGFP) to the N-terminus of one head of the myosin dimer and following the motion with nanometer precision and subsecond resolution. We find the average step size to be 74.1 nm with 9.4 nm (SD) and 0.3 nm (SE). Our measurements demonstrate nanometer localization of single eGFPs, confirm the hand-over-hand model of myosin V procession, and when combined with previous data, suggest that there is a kink in the leading lever arm in the waiting state of myosin V. This kink, or "telemark skier" configuration, may cause strain, which, when released, leads to the powerstroke of myosin, throwing the rear head forward and leading to unidirectional motion. PMID- 15345557 TI - Cardiac length dependence of force and force redevelopment kinetics with altered cross-bridge cycling. AB - We examined the influence of cross-bridge cycling kinetics on the length dependence of steady-state force and the rate of force redevelopment (k(tr)) during Ca(2+)-activation at sarcomere lengths (SL) of 2.0 and 2.3 microm in skinned rat cardiac trabeculae. Cross-bridge kinetics were altered by either replacing ATP with 2-deoxy-ATP (dATP) or by reducing [ATP]. At each SL dATP increased maximal force (F(max)) and Ca(2+)-sensitivity of force (pCa(50)) and reduced the cooperativity (n(H)) of force-pCa relations, whereas reducing [ATP] to 0.5 mM (low ATP) increased pCa(50) and n(H) without changing F(max). The difference in pCa(50) between SL 2.0 and 2.3 microm (Delta pCa(50)) was comparable between ATP and dATP, but reduced with low ATP. Maximal k(tr) was elevated by dATP and reduced by low ATP. Ca(2+)-sensitivity of k(tr) increased with both dATP and low ATP and was unaffected by altered SL under all conditions. Significantly, at equivalent levels of submaximal force k(tr) was faster at short SL or increased lattice spacing. These data demonstrate that the SL dependence of force depends on cross-bridge kinetics and that the increase of force upon SL extension occurs without increasing the rate of transitions between nonforce and force-generating cross-bridge states, suggesting SL or lattice spacing may modulate preforce cross-bridge transitions. PMID- 15345558 TI - The two motor domains of KIF3A/B coordinate for processive motility and move at different speeds. AB - KIF3A/B, a kinesin involved in intraflagellar transport and Golgi trafficking, is distinctive because it contains two nonidentical motor domains. Our hypothesis is that the two heads have distinct functional properties, which are tuned to maximize the performance of the wild-type heterodimer. To test this, we investigated the motility of wild-type KIF3A/B heterodimer and chimaeric KIF3A/A and KIF3B/B homodimers made by splicing the head of one subunit to the rod and tail of the other. The first result is that KIF3A/B is processive, consistent with its transport function in cells. Secondly, the KIF3B/B homodimer moves at twice the speed of the wild-type motor but has reduced processivity, suggesting a trade-off between speed and processivity. Third, the KIF3A/A homodimer moves fivefold slower than wild-type, demonstrating distinct functional differences between the two heads. The heterodimer speed cannot be accounted for by a sequential head model in which the two heads alternate along the microtubule with identical speeds as in the homodimers. Instead, the data are consistent with a coordinated head model in which detachment of the slow KIF3A head from the microtubule is accelerated roughly threefold by the KIF3B head. PMID- 15345559 TI - Alternative N-terminal regions of Drosophila myosin heavy chain tune muscle kinetics for optimal power output. AB - We assessed the influence of alternative versions of a region near the N-terminus of Drosophila myosin heavy chain on muscle mechanical properties. Previously, we exchanged N-terminal regions (encoded by alternative exon 3s) between an embryonic (EMB) isoform and the indirect flight muscle isoform (IFI) of myosin, and demonstrated that it influences solution ATPase rates and in vitro actin sliding velocity. Because each myosin is expressed in Drosophila indirect flight muscle, in the absence of other myosin isoforms, this allows for muscle mechanical and whole organism locomotion assays. We found that exchanging the flight muscle specific exon 3 region into the embryonic isoform (EMB-3b) increased maximum power generation (P(max)) and optimal frequency of power generation (f(max)) threefold and twofold compared to fibers expressing EMB, whereas exchanging the embryonic exon 3 region into the flight muscle isoform (IFI-3a) decreased P(max) and f(max) to approximately 80% of IFI fiber values. Drosophila expressing IFI-3a exhibited a reduced wing beat frequency compared to flies expressing IFI, which optimized power generation from their kinetically slowed flight muscle. However, the slower wing beat frequency resulted in a substantial loss of aerodynamic power as manifest in decreased flight performance of IFI-3a compared to IFI. Thus the N-terminal region is important in tuning myosin kinetics to match muscle speed for optimal locomotory performance. PMID- 15345560 TI - Cross-bridge versus thin filament contributions to the level and rate of force development in cardiac muscle. AB - In striated muscle thin filament activation is initiated by Ca(2+) binding to troponin C and augmented by strong myosin binding to actin (cross-bridge formation). Several lines of evidence have led us to hypothesize that thin filament properties may limit the level and rate of force development in cardiac muscle at all levels of Ca(2+) activation. As a test of this hypothesis we varied the cross-bridge contribution to thin filament activation by substituting 2 deoxy ATP (dATP; a strong cross-bridge augmenter) for ATP as the contractile substrate and compared steady-state force and stiffness, and the rate of force redevelopment (k(tr)) in demembranated rat cardiac trabeculae as [Ca(2+)] was varied. We also tested whether thin filament dynamics limits force development kinetics during maximal Ca(2+) activation by comparing the rate of force development (k(Ca)) after a step increase in [Ca(2+)] with photorelease of Ca(2+) from NP-EGTA to maximal k(tr), where Ca(2+) binding to thin filaments should be in (near) equilibrium during force redevelopment. dATP enhanced steady-state force and stiffness at all levels of Ca(2+) activation. At similar submaximal levels of steady-state force there was no increase in k(tr) with dATP, but k(tr) was enhanced at higher Ca(2+) concentrations, resulting in an extension (not elevation) of the k(tr)-force relationship. Interestingly, we found that maximal k(tr) was faster than k(Ca), and that dATP increased both by a similar amount. Our data suggest the dynamics of Ca(2+)-mediated thin filament activation limits the rate that force develops in rat cardiac muscle, even at saturating levels of Ca(2+). PMID- 15345561 TI - Ca2+ and ionic strength dependencies of S1-ADP binding to actin-tropomyosin troponin: regulatory implications. AB - Skeletal and cardiac muscle contraction are inhibited by the actin-associated complex of tropomyosin-troponin. Binding of Ca(2+) to troponin or binding of ATP free myosin to actin reverses this inhibition. Ca(2+) and ATP-free myosin stabilize different tropomyosin-actin structural arrangements. The position of tropomyosin on actin affects the binding of ATP-free myosin to actin but does not greatly affect myosin-ATP binding. Ca(2+) and ATP-free myosin alter both the affinity of ATP-free myosin for actin and the kinetics of that binding. A parallel pathway model of regulation simulated the effects of Ca(2+) and ATP-free myosin binding on both equilibrium binding of myosin-nucleotide complexes to actin and the general features of ATPase activity. That model was recently shown to simulate the kinetics of myosin-S1 binding but the analysis was limited to a single condition because of the limited data available. We have now measured equilibrium binding and binding kinetics of myosin-S1-ADP to actin at a series of ionic strengths and free Ca(2+) concentrations. The parallel pathway model of regulation is consistent with those data. In that model the interaction between adjacent regulatory complexes fully saturated with Ca(2+) was destabilized and the inactive state of actin was stabilized at high ionic strength. These changes explain the previously observed change in binding kinetics with increasing ionic strength. PMID- 15345562 TI - Organization of Ca2+ release units in excitable smooth muscle of the guinea-pig urinary bladder. AB - Ca(2+) release from internal stores (sarcoplasmic reticulum or SR) in smooth muscles is initiated either via pharmaco-mechanical coupling due to the action of an agonist and involving IP3 receptors, or via excitation-contraction coupling, mostly involving L-type calcium channels in the plasmalemma (DHPRs), and ryanodine receptors (RyRs), or Ca(2+) release channels of the SR. This work focuses attention on the structural basis for the coupling between DHPRs and RyRs in phasic smooth muscle cells of the guinea-pig urinary bladder. Immunolabeling shows that two proteins of the SR: calsequestrin and the RyR, and one protein the plasmalemma, the L-type channel or DHPR, are colocalized with each other within numerous, peripherally located sites located within the caveolar domains. Electron microscopy images from thin sections and freeze-fracture replicas identify feet in small peripherally located SR vesicles containing calsequestrin and distinctive large particles clustered within small membrane areas. Both feet and particle clusters are located within caveolar domains. Correspondence between the location of feet and particle clusters and of RyR- and DHPR-positive foci allows the conclusion that calsequestrin, RyRs, and L-type Ca(2+) channels are associated with peripheral couplings, or Ca(2+) release units, constituting the key machinery involved in excitation-contraction coupling. Structural analogies between smooth and cardiac muscle excitation-contraction coupling complexes suggest a common basic mechanism of action. PMID- 15345563 TI - Contrasting the excited-state dynamics of the photoactive yellow protein chromophore: protein versus solvent environments. AB - Wavelength- and time-resolved fluorescence experiments have been performed on the photoactive yellow protein, the E46Q mutant, the hybrids of these proteins containing a nonisomerizing "locked" chromophore, and the native and locked chromophores in aqueous solution. The ultrafast dynamics of these six systems is compared and spectral signatures of isomerization and solvation are discussed. We find that the ultrafast red-shifting of fluorescence is associated mostly with solvation dynamics, whereas isomerization manifests itself as quenching of fluorescence. The observed multiexponential quenching of the protein samples differs from the single-exponential lifetimes of the chromophores in solution. The locked chromophore in the protein environment decays faster than in solution. This is due to additional channels of excited-state energy dissipation via the covalent and hydrogen bonds with the protein environment. The observed large dispersion of quenching timescales observed in the protein samples that contain the native pigment favors both an inhomogeneous model and an excited-state barrier for isomerization. PMID- 15345564 TI - Incoherent manipulation of the photoactive yellow protein photocycle with dispersed pump-dump-probe spectroscopy. AB - Photoactive yellow protein is the protein responsible for initiating the "blue light vision" of Halorhodospira halophila. The dynamical processes responsible for triggering the photoactive yellow protein photocycle have been disentangled with the use of a novel application of dispersed ultrafast pump-dump-probe spectroscopy, where the photocycle can be started and interrupted with appropriately tuned and timed laser pulses. This "incoherent" manipulation of the photocycle allows for the detailed spectroscopic investigation of the underlying photocycle dynamics and the construction of a fully self-consistent dynamical model. This model requires three kinetically distinct excited-state intermediates, two (ground-state) photocycle intermediates, I(0) and pR, and a ground-state intermediate through which the protein, after unsuccessful attempts at initiating the photocycle, returns to the equilibrium ground state. Also observed is a previously unknown two-photon ionization channel that generates a radical and an ejected electron into the protein environment. This second excitation pathway evolves simultaneously with the pathway containing the one photon photocycle intermediates. PMID- 15345565 TI - Conformational changes in azurin from Pseudomona aeruginosa induced through chemical and physical protocols. AB - Azurin from Pseudomona aeruginosa is a small copper protein with a single tryptophan (Trp) buried in the structure. The Gibbs free energies associated with the folding of holo azurin, calculated monitoring Trp fluorescence and changes in absorbance on the ligand-to-metal band, are different because these techniques probe their local environments, thereby being able to probe different conformational changes. The presence of an intermediate state was observed during the chemical denaturation of the protein. Upon denaturation, a 30-fold increase is observed in the magnitude of the quenching constant of the tryptophan fluorescence by acrylamide, because this residue becomes more accessible to the quencher. Entrapping the protein in sol-gel materials lowers its stability possibly because the solvation properties of the macromolecule are changed. The thermal denaturation of azurin immobilized in a sol-gel monolith is irreversible, which tends to rule out an aggregation mechanism to account for the irreversibility of the denaturation of the protein free in solution. Unlike the Cu(II) ion, the Gd(III) ion accommodates in site B of azurin with high affinity and the folding free energy of Gd-azurin is larger than that of apo azurin. PMID- 15345566 TI - Time-resolved visible and infrared study of the cyano complexes of myoglobin and of hemoglobin I from Lucina pectinata. AB - The dynamics of the ferric CN complexes of the heme proteins Myoglobin and Hemoglobin I from the clam Lucina pectinata upon Soret band excitation is monitored using infrared and broad band visible pump-probe spectroscopy. The transient response in the UV-vis spectral region does not depend on the heme pocket environment and is very similar to that known for ferrous proteins. The main feature is an instantaneous, broad, short-lived absorption signal that develops into a narrower red-shifted Soret band. Significant transient absorption is also observed in the 360-390 nm range. At all probe wavelengths the signal decays to zero with a longest time constant of 3.6 ps. The infrared data on MbCN reveal a bleaching of the C triple bond N stretch vibration of the heme-bound ligand, and the formation of a five-times weaker transient absorption band, 28 cm(-1) lower in energy, within the time resolution of the experiment. The MbC triple bond N stretch vibration provides a direct measure for the return of population to the ligated electronic (and vibrational) ground state with a 3-4 ps time constant. In addition, the CN-stretch frequency is sensitive to the excitation of low frequency heme modes, and yields independent information about vibrational cooling, which occurs on the same timescale. PMID- 15345567 TI - A structural model of polyglutamine determined from a host-guest method combining experiments and landscape theory. AB - Modeling the structure of natively disordered peptides has proved difficult due to the lack of structural information on these peptides. In this work, we use a novel application of the host-guest method, combining folding theory with experiments, to model the structure of natively disordered polyglutamine peptides. Initially, a minimalist molecular model (C(alpha)C(beta)) of CI2 is developed with a structurally based potential and captures many of the folding properties of CI2 determined from experiments. Next, polyglutamine "guest" inserts of increasing length are introduced into the CI2 "host" model and the polyglutamine is modeled to match the resultant change in CI2 thermodynamic stability between simulations and experiments. The polyglutamine model that best mimics the experimental changes in CI2 thermodynamic stability has 1), a beta strand dihedral preference and 2), an attractive energy between polyglutamine atoms 0.75-times the attractive energy between the CI2 host Go-contacts. When free-energy differences in the CI2 host-guest system are correctly modeled at varying lengths of polyglutamine guest inserts, the kinetic folding rates and structural perturbation of these CI2 insert mutants are also correctly captured in simulations without any additional parameter adjustment. In agreement with experiments, the residues showing structural perturbation are located in the immediate vicinity of the loop insert. The simulated polyglutamine loop insert predominantly adopts extended random coil conformations, a structural model consistent with low resolution experimental methods. The agreement between simulation and experimental CI2 folding rates, CI2 structural perturbation, and polyglutamine insert structure show that this host-guest method can select a physically realistic model for inserted polyglutamine. If other amyloid peptides can be inserted into stable protein hosts and the stabilities of these host-guest mutants determined, this novel host-guest method may prove useful to determine structural preferences of these intractable but biologically relevant protein fragments. PMID- 15345568 TI - Probing the self-assembly and the accompanying structural changes of hydrophobin SC3 on a hydrophobic surface by mass spectrometry. AB - The fungal class I hydrophobin SC3 self-assembles into an amphipathic membrane at hydrophilic-hydrophobic interfaces such as the water-air and water-Teflon interface. During self-assembly, the water-soluble state of SC3 proceeds via the intermediate alpha-helical state to the stable end form called the beta-sheet state. Self-assembly of the hydrophobin at the Teflon surface is arrested in the alpha-helical state. The beta-sheet state can be induced at elevated temperature in the presence of detergent. The structural changes of SC3 were monitored by various mass spectrometry techniques. We show that the so-called second loop of SC3 (C39-S72) has a high affinity for Teflon. Binding of this part of SC3 to Teflon was accompanied by the formation of alpha-helical structure and resulted in low solvent accessibility. The solvent-protected region of the second loop extended upon conversion to the beta-sheet state. In contrast, the C-terminal part of SC3 became more exposed to the solvent. The results indicate that the second loop of class I hydrophobins plays a pivotal role in self-assembly at the hydrophilic-hydrophobic interface. Of interest, this loop is much smaller in case of class II hydrophobins, which may explain the differences in their assembly. PMID- 15345569 TI - The role of electrostatic interactions in calmodulin-peptide complex formation. AB - The complex between calmodulin and the calmodulin-binding portion of smMLCKp has been studied. Electrostatic interactions have been anticipated to be important in this system where a strongly negative protein binds a peptide with high positive charge. Electrostatic interactions were probed by varying the pH in the range from 4 to 11 and by charge deletions in CaM and smMLCKp. The change in net charge of CaM from approximately -5 at pH 4.5 to -15 at pH 7.5 leaves the binding constant virtually unchanged. The affinity was also unaffected by mutations in CaM and charge substitutions in the peptide. The insensitivity of the binding constant to pH may seem surprising, but it is a consequence of the high charge on both protein and peptide. At low pH it is further attenuated by a charge regulation mechanism. That is, the protein releases a number of protons when binding the positively charged peptide. We speculate that the role of electrostatic interactions is to discriminate against unbound proteins rather than to increase the affinity for any particular target protein. PMID- 15345570 TI - Stability of the heme environment of the nitric oxide synthase from Staphylococcus aureus in the absence of pterin cofactor. AB - We have used resonance Raman spectroscopy to probe the heme environment of a recently discovered NOS from the pathogenic bacterium Staphylococcus aureus, named SANOS. We detect two forms of the CO complex in the absence of L-arginine, with nu(Fe-CO) at 482 and 497 cm(-1) and nu(C-O) at 1949 and 1930 cm(-1), respectively. Similarly to mammalian NOS, the binding of L-arginine to SANOS caused the formation of a single CO complex with nu(Fe-CO) and nu(C-O) frequencies at 504 and 1,917 cm(-1), respectively, indicating that L-arginine induced an electrostatic/steric effect on the CO molecule. The addition of pterins to CO-bound SANOS modified the resonance Raman spectra only when they were added in combination with L-arginine. We found that (6R) 5,6,7,8 tetra-hydro L-biopterin and tetrahydrofolate were not required for the stability of the reduced protein, which is 5-coordinate, and of the CO complex, which does not change with time to a form with a Soret band at 420 nm that is indicative of a change of the heme proximal coordination. Since SANOS is stable in the absence of added pterin, it suggests that the role of the pterin cofactor in the bacterial NOS may be limited to electron/proton transfer required for catalysis and may not involve maintaining the structural integrity of the protein as is the case for mammalian NOS. PMID- 15345571 TI - Oligomeric beta-structure of the membrane-bound HIV-1 fusion peptide formed from soluble monomers. AB - The human immunodeficiency virus type 1 (HIV-1) fusion peptide serves as a useful model system for understanding viral/target cell fusion, at least to the lipid mixing stage. Previous solid-state NMR studies have shown that the peptide adopts an oligomeric beta-strand structure when associated with a lipid and cholesterol mixture close to that of membranes of host cells of the virus. In this study, this structure was further investigated using four different peptide constructs. In aqueous buffer solution, two of the constructs were primarily monomeric whereas the other two constructs had significant populations of oligomers/aggregates. NMR measurements for all membrane-associated peptide constructs were consistent with oligomeric beta-strand structure. Thus, constructs that are monomeric in solution can be converted to oligomers as a result of membrane association. In addition, samples prepared by very different methods had very similar NMR spectra, which indicates that the beta-strand structure is an equilibrium rather than a kinetically trapped structure. Lipid mixing assays were performed to assess the fusogenicities of the different constructs, and there was not a linear correlation between the solution oligomeric state and fusogenicity. However, the functional assays do suggest that small oligomers may be more fusogenic than either monomers or large aggregates. PMID- 15345572 TI - Using atomic force microscopy to study nucleosome remodeling on individual nucleosomal arrays in situ. AB - In eukaryotes, genomic processes like transcription, replication, repair, and recombination typically require alterations in nucleosome structure on specific DNA regions to operate. ATP-dependent nucleosome remodeling complexes provide a major mechanism for carrying out such alterations in vivo. To learn more about the action of these important complexes, we have utilized an atomic force microscopy in situ technique that permits comparison of the same individual molecules before and after activation of a particular process, in this case nucleosome remodeling. This direct approach was used to look for changes induced by the action of the human Swi-Snf remodeling complex on individual, single-copy mouse mammary tumor virus promoter nucleosomal arrays. Using this technique, we detect a variety of changes on remodeling. Many of these changes are larger in scale than suggested from previous studies and involve a number of DNA-mediated events, including a preference for the removal of a complete turn (80 basepairs) of nucleosomal DNA. The latter result raises the possibility of an unanticipated mode of human Swi-Snf interaction with the nucleosome, namely via the 11-nm histone surface. PMID- 15345573 TI - All-optical constant-force laser tweezers. AB - Optical tweezers are a powerful tool for the study of single biomolecules. Many applications require that a molecule be held under constant tension while its extension is measured. We present two schemes based on scanning-line optical tweezers to accomplish this, providing all-optical alternatives to force-clamp traps that rely on electronic feedback to maintain constant-force conditions for the molecule. In these schemes, a laser beam is rapidly scanned along a line in the focal plane of the microscope objective, effectively creating an extended one dimensional optical potential over distances of up to 8 microm. A position independent lateral force acting on a trapped particle is created by either modulating the laser beam intensity during the scan or by using an asymmetric beam profile in the back focal plane of the microscope objective. With these techniques, forces of up to 2.69 pN have been applied over distances of up to 3.4 microm with residual spring constants of <26.6 fN/microm. We used these techniques in conjunction with a fast position measurement scheme to study the relaxation of lambda-DNA molecules against a constant external force with submillisecond time resolution. We compare the results to predictions from the wormlike chain model. PMID- 15345574 TI - Simultaneous topography and recognition imaging using force microscopy. AB - We present a method for simultaneously recording topography images and localizing specific binding sites with nm positional accuracy by combining dynamic force microscopy with single molecule recognition force spectroscopy. For this we used lysozyme adsorbed to mica, the functionality of which was characterized by enzyme immunoassays. The topography and recognition images were acquired using tips that were magnetically oscillated during scanning and contained antibodies directed against lysozyme. For cantilevers with low Q-factor (approximately 1 in liquid) driven at frequencies below resonance, the surface contact only affected the downward deflections (minima) of the oscillations, whereas binding of the antibody on the tip to lysozyme on the surface only affected the upwards deflections (maxima) of the oscillations. The recognition signals were therefore well separated from the topographic signals, both in space (Delta z approximately 5 nm) and time (approximately 0.1 ms). Topography and recognition images were simultaneously recorded using a specially designed electronic circuit with which the maxima (U(up)) and the minima (U(down)) of each sinusoidal cantilever deflection period were depicted. U(down) was used for driving the feedback loop to record the height (topography) image, and U(up) provided the data for the recognition image. PMID- 15345575 TI - Three-dimensional tracking of single secretory granules in live PC12 cells. AB - Deconvolution wide-field fluorescence microscopy and single-particle tracking were used to study the three-dimensional mobility of single secretory granules in live PC12 cells. Acridine orange-labeled granules were found to travel primarily in random and caged diffusion, whereas only a small fraction of granules traveled in directed fashion. High K(+) stimulation increased significantly the percentage of granules traveling in directed fashion. By dividing granules into the near membrane group (within 1 microm from the plasma membrane) and cytosolic group, we have revealed significant differences between these two groups of granules in their mobility. The mobility of these two groups of granules is also differentially affected by disruption of F-actin, suggesting different mechanisms are involved in the motion of the two groups of granules. Our results demonstrate that combined deconvolution and single-particle tracking may find its application in three-dimensional tracking of long-term motion of granules and elucidating the underlying mechanisms. PMID- 15345576 TI - Micro-volume couette flow sample orientation for absorbance and fluorescence linear dichroism. AB - Linear dichroism (LD) can be used to study the alignment of absorbing chromophores within long molecules. In particular, Couette flow LD has been used to good effect in probing ligand binding to DNA and to fibrous proteins. This technique has been previously limited by large sample requirements. Here we report the design and application of a new micro-volume Couette flow cell that significantly enhances the potential applications of flow LD spectroscopy by reducing the sample requirements for flow linear dichroism to 25 microL (with concentrations such that the absorbance maximum of the sample in a 1-cm pathlength cuvette is approximately 1). The micro-volume Couette cell has also enabled the measurement of fluorescence-detected Couette flow linear dichroism. This new technique enables the orientation of fluorescent ligands to be probed even when their electronic transitions overlap with those of the macromolecule and conversely. The potential of flow-oriented fluorescence dichroism and application of the micro-volume Couette LD cell are illustrated by the collection of data for DNA with minor groove and intercalating ligands: DAPI, Hoechst, and ethidium bromide. As with conventional fluorescence, improved sensitivity compared with absorbance LD is to be expected after instrumentation optimization. PMID- 15345577 TI - The mobility of phytochrome within protonemal tip cells of the moss Ceratodon purpureus, monitored by fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) is a versatile tool for investigating the mobilities of fluorescent molecules in cells. In this article, we show that it is possible to distinguish between freely diffusing and membrane-bound forms of biomolecules involved in signal transduction in living cells. Fluorescence correlation spectroscopy was used to measure the mobility of phytochrome, which plays a role in phototropism and polarotropism in protonemal tip cells of the moss Ceratodon purpureus. The phytochrome was loaded with phycoerythrobilin, which is fluorescent only in the phytochrome-bound state. Confocal laser scanning microscopy was used for imaging and selecting the xy measuring position in the apical zone of the tip cell. Fluorescence correlation was measured at ancient z positions in the cell. Analysis of the diffusion coefficients by nonlinear least square fits showed a subcellular fraction of phytochrome at the cell periphery with a sixfold higher diffusion coefficient than in the core fraction. This phytochrome is apparently bound to the membrane and probably controls the phototropic and polarotropic response. PMID- 15345578 TI - Coordinated behavior of mitochondria in both space and time: a reactive oxygen species-activated wave of mitochondrial depolarization. AB - Reactive oxygen species (ROS) can trigger a transient burst of mitochondrial ROS production via ROS activation of the mitochondrial permeability transition pore (MPTP), a phenomenon termed ROS-induced ROS release (RIRR). The goal of this study was to investigate if the generation of ROS in a discrete region of a cardiomyocyte could serve to propagate RIRR-mediated mitochondrial depolarizations throughout a cell. Our experiments revealed that localized RIRR activated either RIRR-mediated fluctuations in mitochondrial membrane potential (time period: 3-10 min) or a traveling wave of depolarization of the cell's mitochondria (velocity: approximately 5 microm/min). Both phenomena appeared to be mediated by the mitochondrial permeability transition pore and eventually encompassed the majority of the mitochondrial population of both isolated rat and rabbit cardiomyocytes. Furthermore, depolarization was often reversible; the waves of depolarization were then followed by a rapid (approximately 40 microm/min) repolarization wave of the mitochondria. We show that the RIRR can function to communicate the mitochondrial permeability transition from one mitochondrion to another in the isolated adult cardiomyocyte. PMID- 15345579 TI - Pseudopod projection and cell spreading of passive leukocytes in response to fluid shear stress. AB - Recent evidence suggests that circulating leukocytes respond to physiological levels of fluid shear stress. This study was designed to examine the shear stress response of individual leukocytes adhering passively to a glass surface. Human leukocytes were exposed to a step fluid shear stress with amplitude between 0.2 and 4 dyn/cm(2) and duration between 1 and 20 min. The response of the cells was determined in the form of projected cell area measurements by high-resolution observation before, during, and after fluid shear application. All cells selected initially had a round morphology. After application of fluid shear many cells projected pseudopodia and spread on the glass surface. The number of leukocytes responding with pseudopod projection and the extent of cell spreading increased with increasing amplitude and duration of fluid shear stress. Pseudopod projection after exposure to a step fluid shear occurs following a delay that is insensitive to the shear stress amplitude and duration. Leukocytes that did not project pseudopodia and spread in response to low shear stress could be shown to respond to a second shear step of higher amplitude. The spreading response requires an intact actin network and activated myosin molecules. Depleting the cell glycocalyx with protease treatment enhances the spreading response in sheared leukocytes. These results indicate that passive leukocytes respond to fluid shear stress with active pseudopod projection and cell spreading. This behavior may contribute to cell spreading on endothelium and other cells as well as to transendothelial migration of leukocytes in the microcirculation. PMID- 15345580 TI - A Brownian dynamics study of the interaction of Phormidium laminosum plastocyanin with Phormidium laminosum cytochrome f. AB - The interaction of Phormidium laminosum plastocyanin (PC) with P. laminosum cytochrome f (cyt f) was studied using Brownian dynamics (BD) simulations. Few complexes and a low rate of electron transfer were observed for wild-type PC. Increasing the positive electrostatic field on PC by the addition of a Zn(2+) ion in the neighborhood of D44 and D45 on PC (as found in crystal structure of plastocyanin) increased the number of complexes formed and the calculated rates of electron transfer as did PC mutations D44A, D45A, E54A, and E57A. Mutations of charged residues on Phormidium PC and Phormidium cyt f were used to map binding sites on both proteins. In both the presence and absence of the Zn(2+) ion, the following residues on PC interact with cyt f: D44, D45, K6, D79, R93, and K100 that lie in a patch just below H92 and Y88 and D10, E17, and E70 located on the upper portion of the PC molecule. In the absence of the Zn(2+) ion, K6 and K35 on the top of the PC molecule also interact with cyt f. Cyt f residues involved in binding PC, in the absence of the Zn(2+) ion, include E165, D187, and D188 that are located on the small domain of cyt f. The orientation of PC in the complexes was quite random in accordance with NMR results. In the presence of the Zn(2+) ion, K53 and E54 in the lower patch of the PC molecule also interact with cyt f and PC interacts with E86, E95, and E123 on the large domain of cyt f. Also, the orientation of PC in the complexes was much more uniform than in the absence of the Zn(2+) ion. The difference may be due to both the larger electrostatic field and the greater asymmetry of the charge distribution on PC observed in the presence of the Zn(2+) ion. Hydrophobic interactions were also observed suggesting a model of cyt f-PC interactions in which electrostatic forces bring the two molecules together but hydrophobic interactions participate in stabilizing the final electron-transfer-active dock. PMID- 15345581 TI - A mitochondrial oscillator dependent on reactive oxygen species. AB - We describe a unique mitochondrial oscillator that depends on oxidative phosphorylation, reactive oxygen species (ROS), and mitochondrial inner membrane ion channels. Cell-wide synchronized oscillations in mitochondrial membrane potential (Delta Psi(m)), NADH, and ROS production have been recently described in isolated cardiomyocytes, and we have hypothesized that the balance between superoxide anion efflux through inner membrane anion channels and the intracellular ROS scavenging capacity play a key role in the oscillatory mechanism. Here, we formally test the hypothesis using a computational model of mitochondrial energetics and Ca(2+) handling including mitochondrial ROS production, cytoplasmic ROS scavenging, and ROS activation of inner membrane anion flux. The mathematical model reproduces the period and phase of the observed oscillations in Delta Psi(m), NADH, and ROS. Moreover, we experimentally verify model predictions that the period of the oscillator can be modulated by altering the concentration of ROS scavengers or the rate of oxidative phosphorylation, and that the redox state of the glutathione pool oscillates. In addition to its role in cellular dysfunction during metabolic stress, the period of the oscillator can be shown to span a wide range, from milliseconds to hours, suggesting that it may also be a mechanism for physiological timekeeping and/or redox signaling. PMID- 15345582 TI - Multiphysics simulation of left ventricular filling dynamics using fluid structure interaction finite element method. AB - To relate the subcellular molecular events to organ level physiology in heart, we have developed a three-dimensional finite-element-based simulation program incorporating the cellular mechanisms of excitation-contraction coupling and its propagation, and simulated the fluid-structure interaction involved in the contraction and relaxation of the human left ventricle. The FitzHugh-Nagumo model and four-state model representing the cross-bridge kinetics were adopted for cellular model. Both ventricular wall and blood in the cavity were modeled by finite element mesh. An arbitrary Lagrangian Eulerian finite element method with automatic mesh updating has been formulated for large domain changes, and a strong coupling strategy has been taken. Using electrical analog of pulmonary circulation and left atrium as a preload and the windkessel model as an afterload, dynamics of ventricular filling as well as ejection was simulated. We successfully reproduced the biphasic filling flow consisting of early rapid filling and atrial contraction similar to that reported in clinical observation. Furthermore, fluid-structure analysis enabled us to analyze the wave propagation velocity of filling flow. This simulator can be a powerful tool for establishing a link between molecular abnormality and the clinical disorder at the macroscopic level. PMID- 15345583 TI - Microscopic Kinetics of DNA Translocation through synthetic nanopores. AB - We have previously demonstrated that a nanometer-diameter pore in a nanometer thick metal-oxide-semiconductor-compatible membrane can be used as a molecular sensor for detecting DNA. The prospects for using this type of device for sequencing DNA are avidly being pursued. The key attribute of the sensor is the electric field-induced (voltage-driven) translocation of the DNA molecule in an electrolytic solution across the membrane through the nanopore. To complement ongoing experimental studies developing such pores and measuring signals in response to the presence of DNA, we conducted molecular dynamics simulations of DNA translocation through the nanopore. A typical simulated system included a patch of a silicon nitride membrane dividing water solution of potassium chloride into two compartments connected by the nanopore. External electrical fields induced capturing of the DNA molecules by the pore from the solution and subsequent translocation. Molecular dynamics simulations suggest that 20-basepair segments of double-stranded DNA can transit a nanopore of 2.2 x 2.6 nm(2) cross section in a few microseconds at typical electrical fields. Hydrophobic interactions between DNA bases and the pore surface can slow down translocation of single-stranded DNA and might favor unzipping of double-stranded DNA inside the pore. DNA occluding the pore mouth blocks the electrolytic current through the pore; these current blockades were found to have the same magnitude as the blockade observed when DNA transits the pore. The feasibility of using molecular dynamics simulations to relate the level of the blocked ionic current to the sequence of DNA was investigated. PMID- 15345584 TI - Stabilization of IFN-gamma mRNA by MAPK p38 in IL-12- and IL-18-stimulated human NK cells. AB - The rapid induction of interferon-gamma (IFN-gamma) by innate cytokines such as interleukin 12 (IL-12) and IL-18 is critical for immunity against infectious pathogens. We investigated the molecular mechanisms underlying this response. IL 12 and IL-18 rapidly and synergistically induced the secretion of IFN-gamma by freshly purified human peripheral blood lymphocytes. At early time points, IFN gamma was expressed almost exclusively by natural killer cells and in both CD56bright and CD56dim subpopulations. Mitogen-activated protein kinase p38 was activated strongly by IL-18 and weakly by IL-12 in natural killer cells but was not activated by either cytokine in T cells. The expression of IFN-gamma mRNA and protein was dose-dependently blocked by SB203580, a specific inhibitor of mitogen activated protein kinase p38, which also caused a dramatic destabilization of IFN gamma mRNA. The 3' untranslated region (UTR) of IFN-gamma mRNA conferred p38 responsiveness to a heterologous reporter mRNA. Therefore, the synergistic induction of IFN-gamma by IL-12 and IL-18 in natural killer cells is mediated at least in part by p38-dependent and 3' UTR-mediated stabilization of IFN-gamma mRNA. PMID- 15345585 TI - Vascular endothelial growth factor regulation of Weibel-Palade-body exocytosis. AB - Vascular endothelial growth factor (VEGF) not only regulates angiogenesis, vascular permeability, and vasodilation but also promotes vascular inflammation. However, the molecular basis for the proinflammatory effects of VEGF is not understood. We now show that VEGF activates endothelial cell exocytosis of Weibel Palade bodies, releasing vasoactive substances capable of causing vascular thrombosis and inflammation. VEGF triggers endothelial exocytosis in part through calcium and phospholipase C-gamma (PLC-gamma) signal transduction. However, VEGF also modulates endothelial cell exocytosis by activating endothelial nitric oxide synthase (eNOS) production of nitric oxide (NO), which nitrosylates N ethylmaleimide sensitive factor (NSF) and inhibits exocytosis. Thus, VEGF plays a dual role in regulating endothelial exocytosis, triggering pathways that both promote and inhibit endothelial exocytosis. Regulation of endothelial exocytosis may explain part of the proinflammatory effects of VEGF. PMID- 15345586 TI - CD1d-unrestricted human NKT cells release chemokines upon Fas engagement. AB - Attempts at inducing allograft immune privilege by enforced Fas ligand expression have shown accelerated rejection mediated by neutrophils. While it has been proposed that Fas ligand was directly chemotactic toward neutrophils, several lines of evidence argue for an indirect recruitment mechanism. This question was addressed by using in vitro migration assays that used highly purified human leukocyte subsets. Granulocytes did not migrate in response to Fas engagement and required the presence of T cells expressing several natural killer (NK) cell markers. These rare CD8 memory T cells expressed T and NK cell markers and were not restricted to CD1d, showing that they are distinct from conventional natural killer T (NKT) cells. These cells were able to kill both NK-sensitive and insensitive targets and secreted several CC and CXC chemokines active toward granulocytes, monocytes, and NK cells upon Fas engagement. Chemotactic factor release depended on caspase activity, in the absence of NKT cell apoptosis. The ability of CD1d-unrestricted NKT cells to recruit innate immune system cells might play a role in cancer cell eradication and contribute to inflammatory diseases. PMID- 15345587 TI - Expression of hepcidin is down-regulated in TfR2 mutant mice manifesting a phenotype of hereditary hemochromatosis. AB - Transferrin receptor 2 (TfR2) is a membrane glycoprotein that mediates cellular iron uptake from holotransferrin. Homozygous mutations of this gene cause one form of hereditary hemochromatosis in humans. We recently reported that homozygous TfR2(Y245X) mutant mice, which correspond to the TfR2(Y250X) mutation in humans, showed a phenotype similar to hereditary hemochromatosis. In this study, we further analyzed the phenotype as well as iron-related gene expression in these mice by comparing the TfR2-mutant and wild-type siblings. Northern blot analyses showed that the levels of expression of hepcidin mRNA in the liver were generally lower, whereas those of duodenal DMT1, the main transporter for uptake of dietary iron, were higher in the TfR2-mutant mice as compared to the wild-type siblings. Expression of hepcidin mRNA in the TfR2 mutant mice remained low even after intraperitoneal iron loading. In isolated hepatocytes from both wild-type and TfR2 mutant mice, interleukin-6 and lipopolysaccharide each induced expression of hepcidin mRNA. These results suggest that up-regulation of hepcidin expression by inflammatory stimuli is independent of TfR2 and that TfR2 is upstream of hepcidin in the regulatory pathway of body iron homeostasis. PMID- 15345588 TI - Interleukin-15 induces IL-12 receptor beta1 gene expression through PU.1 and IRF 3 by targeting chromatin remodeling. AB - Interleukin-12 receptor beta1 (IL12RB1) is expressed on a variety of immune cells, including T and natural killer (NK) cells and macrophages, and is involved in innate and adaptive immune responses. Levels of IL12RB1 mRNA are dynamically regulated by various cytokines, including interferon-gamma (IFN-gamma) and IL-15. To reveal the regulatory mechanisms governing IL12RB1 gene expression, we analyzed the transcriptional regulatory region of the mouse IL12RB1 gene. Promoter analyses in a mouse macrophage cell line, RAW264.7, revealed that the 2508-bp region upstream of the transcriptional start site is sufficient for the full transcriptional activation of the IL12RB1 gene by IFN-gamma or IL-15. Analyses of the deletion mutants revealed critical roles of IRE/ISRE and ETS/PU.1 elements, to which IRF3 and PU.1, respectively, bound. Notably, chromatin immunoprecipitation (ChIP) assays revealed IL-15 rapidly induced histone H3 acetylation at the IL12RB1 promoter. Consistently, IL-15, as a histone deacetylase inhibitor, synergistically enhanced IL12RB1 gene expression and promoter activation by IFN-gamma through increased protein binding to ETS/PU.1 and IRE/ISRE sites. Additionally, IL12RB1 promoter activation by IFN-gamma was enhanced by the coexpression of a coactivator protein, CBP. Thus, IL-15 induces chromatin remodeling of the IL12RB1 gene promoter, increasing IL12RB1 mRNA expression in synergy with IFN-gamma through the recruitment of PU.1 and IRF3. PMID- 15345589 TI - Gene expression profiling in follicular lymphoma to assess clinical aggressiveness and to guide the choice of treatment. AB - Follicular lymphoma (FL) is a disease characterized by a long clinical course marked by frequent relapses that vary in clinical aggressiveness over time. Therefore, the main dilemma at each relapse is the choice for the most effective treatment for optimal disease control and failure-free survival while at the same time avoiding overtreatment and harmful side effects. The selection for more aggressive treatment is currently based on histologic grading and clinical criteria; however, in up to 30% of all cases these methods prove to be insufficient. Using supervised classification on a training set of paired samples from patients who experienced either an indolent or aggressive disease course, a gene expression profile of 81 genes was established that could, with an accuracy of 100%, distinguish low-grade from high-grade disease. This profile accurately classified 93% of the FL samples in an independent validation set. Most important, in a third series of FL cases where histologic grading was ambiguous, precluding meaningful morphologic guidance, the 81-gene profile shows a classification accuracy of 94%. The FL stratification profile is a more reliable marker of clinical behavior than the currently used histologic grading and clinical criteria and may provide an important alternative to guide the choice of therapy in patients with FL both at presentation and at relapse. PMID- 15345590 TI - Increased circulating hematopoietic and endothelial progenitor cells in the early phase of acute myocardial infarction. AB - Endothelial progenitor cell (EPC) mobilization has been reported following tissue damage, whereas no data are available regarding the mobilization of hematopoietic progenitor cells (HPCs). We performed the phenotypic and functional analysis of circulating CD34+ progenitor cells in patients with acute myocardial infarction (AMI), assessed from admission up to 60 days, in patients with stable angina pectoris (SA), and in healthy controls (CTRLs). In patients with AMI at admission (T0), the number of circulating CD34+ cells was higher (P < .001) than in CTRLs and became comparable with CTRLs within 60 days. Both the number of CD34+ cells coexpressing CD33, CD38, or CD117 and the number of HPCs was higher (P < .02 for all) in patients with AMI at T0 than in CTRLs, as was the number of hematopoietic colonies (P < .03). Patients with AMI (T0) had a significantly increased number of CD34+ vascular endothelial growth factor receptor 2-positive (VEGFR-2+) cells (P < .002) with respect to CTRLs, including CD34(+) CD133(+)VEGFR-2+ and CD34+ CD117(+)VEGFR-2+ EPCs. The number of endothelial colonies was higher in patients with AMI (T0) than in CTRLs (P < .05). No significant difference was documented between patients with SA and CTRLs. Spontaneous mobilization of both HPCs and EPCs occurs within a few hours from the onset of AMI and is detectable until 2 months. PMID- 15345591 TI - Receptor-binding residues lie in central regions of Duffy-binding-like domains involved in red cell invasion and cytoadherence by malaria parasites. AB - Erythrocyte invasion by malaria parasites and cytoadherence of Plasmodium falciparum-infected erythrocytes to host capillaries are 2 key pathogenic mechanisms in malaria. The receptor-binding domains of erythrocyte-binding proteins (EBPs) such as Plasmodium falciparum EBA-175, which mediate invasion, and P falciparum erythrocyte membrane protein 1 (PfEMP-1) family members, which are encoded by var genes and mediate cytoadherence, have been mapped to conserved cysteine-rich domains referred to as Duffy-binding-like (DBL) domains. Here, we have mapped regions within DBL domains from EBPs and PfEMP-1 that contain receptor-binding residues. Using biochemical and molecular methods we demonstrate that the receptor-binding residues of parasite ligands that bind sialic acid on glycophorin A for invasion as well as complement receptor-1 and chondroitin sulfate A for cytoadherence map to central regions of DBL domains. In contrast, binding to intercellular adhesion molecule 1 (ICAM-1) requires both the central and terminal regions of DBLbetaC2 domains. Determination of functional regions within DBL domains is the first step toward understanding the structure-function bases for their interaction with diverse host receptors. PMID- 15345592 TI - Detection of BCR-ABL kinase mutations in CD34+ cells from chronic myelogenous leukemia patients in complete cytogenetic remission on imatinib mesylate treatment. AB - The BCR-ABL kinase inhibitor imatinib mesylate induces complete cytogenetic response (CCR) in a high proportion of chronic myelogenous leukemia (CML) patients. However, patients in CCR usually demonstrate evidence of residual BCR ABL-positive progenitors. The mechanisms underlying persistence of small numbers of malignant progenitors in imatinib-sensitive patients are unclear. BCR-ABL kinase domain mutations affecting drug binding can lead to secondary resistance to imatinib. We show here that kinase mutations could be detected in CD34+ cells isolated from CML patients in CCR on imatinib. Most mutations seen have not been reported in previous clinical studies. Interestingly, several of the involved amino acid positions have been implicated in an in vitro mutagenesis screen. These BCR-ABL mutations were associated with varying levels of imatinib resistance. Two of 5 patients in whom mutations were detected on initial evaluation have relapsed. In addition, 4 patients in whom mutations were not initially detected, but with rising BCR-ABL mRNA levels on quantitative polymerase chain reaction (Q-PCR) analysis, had mutations detected on follow-up evaluation. We conclude that BCR-ABL kinase mutations can be detected in CD34+ cells from CML patients in CCR on imatinib, may contribute to persistence of small populations of malignant progenitors, and could be a potential source of relapse. PMID- 15345593 TI - Identification of a novel activating mutation (Y842C) within the activation loop of FLT3 in patients with acute myeloid leukemia (AML). AB - Fms-like tyrosine kinase 3 (FLT3) receptor mutations as internal tandem duplication (ITD) or within the kinase domain are detected in up to 35% of patients with acute myeloid leukemia (AML). N-benzoyl staurosporine (PKC412), a highly effective inhibitor of mutated FLT3 receptors, has significant antileukemic efficacy in patients with FLT3-mutated AML. Mutation screening of FLT3 exon 20 in AML patients (n = 110) revealed 2 patients with a novel mutation (Y842C) within the highly conserved activation loop of FLT3. FLT3-Y842C transfected 32D cells showed constitutive FLT3 tyrosine phosphorylation and interleukin 3 (IL-3)-independent growth. Treatment with PKC412 led to inhibition of proliferation and apoptotic cell death. Primary AML blasts bearing FLT3-Y842C mutations showed constitutive FLT3 and signal transducer and activator of transcription 5 (STAT-5) tyrosine phosphorylation. Ex vivo PKC412 treatment of primary blasts resulted in suppression of constitutive FLT3 and STAT-5 activation and apoptotic cell death. Inspection of the FLT3 structure revealed that Y842 is the key residue in regulating the switch from the closed to the open (= active) conformation of the FLT3 activation loop. Overall, our data suggest that mutations at Y842 represent a significant new activating mutation in AML blasts. Since FLT3 tyrosine kinase inhibitors (TKIs) such as PKC412 are currently being investigated in clinical trials in AML, extended sequence analysis of FLT3 may be helpful in defining the spectrum of TKI-sensitive FLT3 mutations in AML. PMID- 15345594 TI - The adaptor protein 3BP2 associates with VAV guanine nucleotide exchange factors to regulate NFAT activation by the B-cell antigen receptor. AB - Engagement of the B-cell antigen receptor (BCR) activates kinases of the Src and Syk families and signaling complexes assembled by adaptor proteins, which dictate B-cell fate and function. The adaptor 3BP2/SH3BP2, an Abl Src homology domain 3 (SH3)-binding and Syk-kinases interacting protein, exhibits positive regulatory roles in T, natural killer (NK), and basophilic cells. However, its involvement in BCR signaling is completely unknown. Here we show that 3BP2 is tyrosine phosphorylated following BCR aggregation on B lymphoma cells, and that 3BP2 is a substrate for Syk and Fyn, but not Btk. To further explore the function of 3BP2 in B cells, we screened a yeast 2-hybrid B-lymphocyte library and found 3BP2 as a binding partner of Vav proteins. The interaction between 3BP2 and Vav proteins involved both constitutive and inducible mechanisms. 3BP2 also interacted with other components of the BCR signaling pathway, including Syk and phospholipase C gamma (PLC-gamma). Furthermore, overexpression and RNAi blocking experiments showed that 3BP2 regulated BCR-mediated activation of nuclear factor of activated T cells (NFATs). Finally, evidence was provided that 3BP2 functionally cooperates with Vav proteins and Rho GTPases to activate NFATs. Our results show that 3BP2 may regulate BCR-mediated gene activation through Vav proteins. PMID- 15345595 TI - Transition of late-stage effector T cells to CD27+ CD28+ tumor-reactive effector memory T cells in humans after adoptive cell transfer therapy. AB - In humans, the pathways of memory T-cell differentiation remain poorly defined. Recently, adoptive cell transfer (ACT) of tumor-reactive T lymphocytes to metastatic melanoma patients after nonmyeloablative chemotherapy has resulted in persistence of functional, tumor-reactive lymphocytes, regression of disease, and induction of melanocyte-directed autoimmunity in some responding patients. In the current study, longitudinal phenotypic analysis was performed on melanoma antigen specific CD8+ T cells during their transition from in vitro cultured effector cells to long-term persistent memory cells following ACT to 6 responding patients. Tumor-reactive T cells used for therapy were generally late-stage effector cells with a CD27Lo CD28Lo CD45RA- CD62 ligand- (CD62L-) CC chemokine receptor 7- (CCR7-) interleukin-7 receptor alphaLo (IL-7RalphaLo) phenotype. After transfer, rapid up-regulation and continued expression of IL-7Ralpha in vivo suggested an important role for IL-7R in immediate and long-term T-cell survival. Although the tumor antigen-specific T-cell population contracted between 1 and 4 weeks after transfer, stable numbers of CD27+)CD28+ tumor reactive T cells were maintained, demonstrating their contribution to the development of long-term, melanoma-reactive memory CD8+ T cells in vivo. At 2 months after transfer, melanoma-reactive T cells persisted at high levels and displayed an effector memory phenotype, including a CD27+ CD28+ CD62L- CCR7- profile, which may explain in part their ability to mediate tumor destruction. PMID- 15345596 TI - Soluble factors elaborated by human brain endothelial cells induce the concomitant expansion of purified human BM CD34+CD38- cells and SCID-repopulating cells. AB - The CD34(+)CD38- phenotype identifies a population in the bone marrow that is enriched in the steady state for hematopoietic stem cells (HSCs). Following ex vivo culture of CD34(+) cells, HSC content is difficult to measure since committed CD34(+)CD38+ progenitors down-regulate CD38 surface expression during culture. In this study, we sought to define the phenotype of human HSCs following ex vivo culture under conditions that support the expansion of human cells capable of repopulating non-obese diabetic/severe combined immunodeficiency (SCID)-repopulating cells (SRCs). Contact coculture of fluorescence-activated cell sorter (FACS)-sorted bone marrow (BM) CD34(+)CD38- cells with human brain endothelial cells (HUBECs) supported a 4.4-fold increase in CD34(+)CD38- cells with a concordant 3.6-fold increase in SRCs over 7 days. Noncontact HUBEC cultures and the addition of thrombopoietin, stem cell factor (SCF), and macrophage colony stimulating factor I receptor (Fms)-like tyrosine kinase 3 (Flt 3) ligand supported further increases in CD34(+)CD38- cells (6.4-fold and 13.1 fold), which correlated with significant increases in SRC activity. Moreover, cell-sorting studies performed on HUBEC-cultured populations demonstrated that SRCs were significantly enriched within the CD34(+)CD38- subset compared with the CD34(-)CD38- population after culture. These results indicate that human HSCs can be identified and characterized by phenotype following expansion culture. These studies also demonstrate that HUBEC-elaborated soluble factors mediate a unique and potent expansion of human HSCs. PMID- 15345597 TI - Patients with acute myeloid leukemia and an activating mutation in FLT3 respond to a small-molecule FLT3 tyrosine kinase inhibitor, PKC412. AB - Leukemic cells from 30% of patients with acute myeloid leukemia (AML) have an activating mutation in the FLT3 (fms-like tyrosine kinase) gene, which represents a target for drug therapy. We treated 20 patients, each with mutant FLT3 relapsed/refractory AML or high-grade myelodysplastic syndrome and not believed to be candidates for chemotherapy, with an FLT3 tyrosine kinase inhibitor, PKC412 (N-benzoylstaurosporine), at a dose of 75 mg 3 times daily by mouth. The drug was generally well tolerated, although 2 patients developed fatal pulmonary events of unclear etiology. The peripheral blast count decreased by 50% in 14 patients (70%). Seven patients (35%) experienced a greater than 2-log reduction in peripheral blast count for at least 4 weeks (median response duration, 13 weeks; range, 9-47 weeks); PKC412 reduced bone marrow blast counts by 50% in 6 patients (2 of these to < 5%). FLT3 autophosphorylation was inhibited in most of the Corresponding patients, indicating in vivo target inhibition at the dose schedule used in this study. PKC412 is an oral tyrosine kinase inhibitor with clinical activity in patients with AML whose blasts have an activating mutation of FLT3, suggesting potential use in combination with active agents, such as chemotherapy. PMID- 15345598 TI - Stromal cell-derived factor-1alpha/CXCL12-induced chemotaxis of T cells involves activation of the RasGAP-associated docking protein p62Dok-1. AB - Events mediating stromal cell-derived factor-1 (SDF-1alpha/CXCL12) chemotaxis of lymphocytes are not completely known. We evaluated intracellular signaling through RasGAP-associated protein p62Dok-1 (downstream of tyrosine kinase [Dok 1]) and associated proteins. SDF-1alpha/CXCL12 stimulated Dok-1 tyrosine phosphorylation and association with RasGAP, adaptor protein p46Nck, and Crk-L in Jurkat T cells. The phosphorylation of Dok-1 was blocked by pretreatment of cells with the src kinase inhibitor PP2. Src kinase family member Lck was implicated. SDF-1alpha/CXCL12 did not phosphorylate Dok-1 in J.CaM1.6 cells, a Jurkat derivative not expressing Lck, but did phosphorylate Dok-1 in J.CaM1.6 cells expressing Lck. SDF-1alpha/CXCL12 induced the tyrosine phosphorylation of Pyk2 and the association of Pyk2 with zeta chain-associated protein-70 kilodaltons (Zap-70) and Vav. SDF-1alpha/CXCL12 enhanced the association of RasGAP with Pyk2. CXCR4-expressing NIH3T3 and Baf3 cells transfected with full-length Dok-1 cDNA were suppressed in their responses to SDF-1alpha/CXCL12-induced chemotaxis; mitogen-activated protein (MAP) kinase activity was also decreased. Chemotaxis to SDF-1/CXCL12 was significantly enhanced in Dok-1(-/-) CD4+ and CD8+ splenic T cells. These results implicate Dok-1, Nck, Crk-L, and Src kinases-especially Lck, Pyk2, Zap-70, Vav, and Ras-GAP-in intracellular signaling by SDF-1alpha/CXCL12, and they suggest that Dok-1 plays an important role in SDF-1alpha/CXCL12-induced chemotaxis in T cells. PMID- 15345599 TI - Temporal relation between depression and cognitive impairment in old age: prospective population based study. AB - OBJECTIVE: To examine the temporal relation between depression and cognitive impairment in old age. DESIGN: Prospective, population based study with four years of follow up. SETTING: City of Leiden, the Netherlands. PARTICIPANTS: 500 people aged 85 years at recruitment. MAIN OUTCOME MEASURES: Annual assessments of depressive symptoms (15 item geriatric depression scale), global cognitive function (mini-mental state examination), attention (Stroop test), processing speed (letter digit coding test), and immediate and delayed recall (12 word learning test). RESULTS: At 85 years old, participants' depressive symptoms and cognitive impairment were highly significantly correlated (P < 0.001). During follow up, an accelerated annual increase of depressive symptoms was associated with impaired attention (0.08 points (95% confidence interval 0.01 to 0.16)), immediate recall (0.17 points (0.09 to 0.25)), and delayed recall (0.10 points (0.02 to 0.18)) at baseline. In contrast, depressive symptoms at baseline were not related to an accelerated cognitive decline during follow up (P > 0.05). CONCLUSION: Caregivers should be aware of the development of depressive symptoms when cognitive impairment is present. However, the presence of depression only does not increase the risk of cognitive decline. PMID- 15345600 TI - Re-engineering systems for the treatment of depression in primary care: cluster randomised controlled trial. AB - OBJECTIVE: To test the effectiveness of an evidence based model for management of depression in primary care with support from quality improvement resources. DESIGN: Cluster randomised controlled trial. SETTING: Five healthcare organisations in the United States and 60 affiliated practices. PATIENTS: 405 patients, aged > or = 18 years, starting or changing treatment for depression. INTERVENTION: Care provided by clinicians, with staff providing telephone support under supervision from a psychiatrist. MAIN OUTCOME MEASURES: Severity of depression at three and six months (Hopkins symptom checklist-20): response to treatment (> or = 50% decrease in scores) and remission (score of < 0.5). RESULTS: At six months, 60% (106 of 177) of patients in intervention practices had responded to treatment compared with 47% (68 of 146) of patients in usual care practices (P = 0.02). At six months, 37% of intervention patients showed remission compared with 27% for usual care patients (P = 0.014). 90% of intervention patients rated their depression care as good or excellent at six months compared with 75% of usual care patients (P = 0.0003). CONCLUSION: Resources such as quality improvement programmes can be used effectively in primary care to implement evidence based management of depression and improve outcomes for patients with depression. PMID- 15345601 TI - Preventing the spread of MRSA. PMID- 15345602 TI - Health care in Armenia. PMID- 15345603 TI - Treating hypertension with guidelines in general practice. PMID- 15345604 TI - Bioterrorism and compulsory vaccination. PMID- 15345605 TI - Stopping clinical trials early. PMID- 15345606 TI - Study indicates nine risk factors explain most heart attacks. PMID- 15345607 TI - UK audit will review pathologists' reports on adult autopsies. PMID- 15345608 TI - Slow progress on sanitation puts 2.6 billion people at risk. PMID- 15345613 TI - Pakistan medical association warns of potential rise in hepatitis. PMID- 15345615 TI - Figures show more Americans do not have insurance. PMID- 15345618 TI - Nearly a third of US adults have high blood pressure. PMID- 15345621 TI - South Africa and Britain reach agreement to curb poaching of healthcare staff. PMID- 15345622 TI - UN warns that aid for Sudan is "grossly underfunded". PMID- 15345623 TI - United States prepares for another flu pandemic. PMID- 15345624 TI - Counterfeits of impotence drug appear in the United Kingdom. PMID- 15345626 TI - Isolation measures in the hospital management of methicillin resistant Staphylococcus aureus (MRSA): systematic review of the literature. AB - OBJECTIVE: To evaluate the evidence for the effectiveness of isolation measures in reducing the incidence of methicillin resistant Staphylococcus aureus (MRSA) colonisation and infection in hospital inpatients. DESIGN: Systematic review of published articles. DATA SOURCES: Medline, Embase, CINAHL, Cochrane Library, System for Information on Grey Literature in Europe (SIGLE), and citation lists (1966-2000). REVIEW METHODS: Articles reporting MRSA related outcomes and describing an isolation policy were selected. No quality restrictions were imposed on studies using isolation wards or nurse cohorting. Other studies were included if they were prospective or employed planned comparisons of retrospective data. RESULTS: 46 studies were accepted; 18 used isolation wards, nine used nurse cohorting, and 19 used other isolation policies. Most were interrupted time series, with few planned formal prospective studies. All but one reported multiple interventions. Consideration of potential confounders, measures to prevent bias, and appropriate statistical analysis were mostly lacking. No conclusions could be drawn in a third of studies. Most others provided evidence consistent with a reduction of MRSA acquisition. Six long interrupted time series provided the strongest evidence. Four of these provided evidence that intensive control measures including patient isolation were effective in controlling MRSA. In two others, isolation wards failed to prevent endemic MRSA. CONCLUSION: Major methodological weaknesses and inadequate reporting in published research mean that many plausible alternative explanations for reductions in MRSA acquisition associated with interventions cannot be excluded. No well designed studies exist that allow the role of isolation measures alone to be assessed. None the less, there is evidence that concerted efforts that include isolation can reduce MRSA even in endemic settings. Current isolation measures recommended in national guidelines should continue to be applied until further research establishes otherwise. PMID- 15345627 TI - Golden rules. PMID- 15345628 TI - Women's reasons for not participating in follow up visits before starting short course antiretroviral prophylaxis for prevention of mother to child transmission of HIV: qualitative interview study. AB - OBJECTIVE: To find out why pregnant women who receive HIV-1 positive test results and are offered short course antiretroviral prophylaxis to prevent transmission of HIV from mother to child do not participate in necessary follow up visits before starting prophylaxis. DESIGN: Qualitative interview study. SETTING: A programme aiming to prevent transmission of HIV from mother to child at a public antenatal clinic in Abidjan, Cote d'Ivoire. PARTICIPANTS: Purposive sample of 27 women who had received HIV-1 positive test results and were invited to return for monthly follow up visits before starting prophylaxis with zidovudine at 36 weeks' gestation, but who had either refused or discontinued the visits. None of the women started prophylaxis. RESULTS: Most of the women explained their non participation in follow up visits by referring to negative experiences that they had had while interacting with programme staff or to their views about the programme. Additional reasons concerned their disbelief of HIV positive test results and personal factors. CONCLUSIONS: Difficulties experienced by women during their contacts with staff working on the prevention programme and negative views that they have about the programme can contribute to their non participation in prophylaxis. Training and supervision of programme staff may increase the likelihood of positive interactions between staff and clients, thereby facilitating women's participation in preventing transmission of HIV from mother to child. Outreach and mobilisation in communities that are served by prevention programmes may complement these measures at programme level by contributing to increased social support for women's efforts to prevent transmission of HIV from mother to child. PMID- 15345629 TI - Relation between online "hit counts" and subsequent citations: prospective study of research papers in the BMJ. PMID- 15345630 TI - Recent developments in Bell's palsy. PMID- 15345631 TI - Rare causes of haemoptysis in suspected pulmonary embolism. PMID- 15345632 TI - Sexual problems associated with infertility, pregnancy, and ageing. PMID- 15345633 TI - Understanding health care in the south Caucasus: examples from Armenia. PMID- 15345634 TI - Consent to the publication of patient information. PMID- 15345635 TI - Guidelines from the British Hypertension Society: cholesterol values seem to diverge. PMID- 15345636 TI - Guidelines from the British Hypertension Society: life in the real world may not allow recommendations to be implemented. PMID- 15345637 TI - Guidelines from the British Hypertension Society: BHS is set to bankrupt NHS. PMID- 15345638 TI - Guidelines from the British Hypertension Society: numbers are missing. PMID- 15345639 TI - Guidelines from the British Hypertension Society: is hypertension really a disease? PMID- 15345640 TI - NICE clinical guidelines: maybe health economists should participate in guideline development. PMID- 15345641 TI - NICE clinical guidelines: account of guideline development was inadequate. PMID- 15345642 TI - NICE clinical guidelines: health economics must engage with complexity of issues. PMID- 15345643 TI - Eradication of MRSA by "ring fencing" orthopaedic beds: stand alone orthopaedic units may be way forward in reducing MRSA. PMID- 15345644 TI - "Serious" and "severe" adverse drug reactions need defining. PMID- 15345645 TI - Stigma of AIDS needs to be overcome. PMID- 15345646 TI - Shouldn't patients decide who should access their records? PMID- 15345647 TI - Burns rehabilitation is more than skin deep. PMID- 15345648 TI - How protective is the working time directive? Two doctors mull over personal horror stories. PMID- 15345649 TI - How protective is the working time directive? New Zealand is still dealing with the issues 20 years on... PMID- 15345650 TI - How protective is the working time directive?... but Australia may have a way forward. PMID- 15345651 TI - Transgenic expression of Bcl-2 modulates energy metabolism, prevents cytosolic acidification during ischemia, and reduces ischemia/reperfusion injury. AB - The antiapoptotic protein Bcl-2 is targeted to the mitochondria, but it is uncertain whether Bcl-2 affects only myocyte survival after ischemia, or whether it also affects metabolic functions of mitochondria during ischemia. Hearts from mice overexpressing human Bcl-2 and from their wild-type littermates (WT) were subjected to 24 minutes of global ischemia followed by reperfusion. During ischemia, the decrease in pH(i) and the initial rate of decline in ATP were significantly reduced in Bcl-2 hearts compared with WT hearts (P<0.05). The reduced acidification during ischemia was dependent on the activity of mitochondrial F1F0-ATPase. In the presence of oligomycin (Oligo), an F1F0-ATPase inhibitor, the decrease in pH(i) was attenuated in WT hearts, but in Bcl-2 hearts, Oligo had no additional effect on pH(i) during ischemia. Likewise, addition of Oligo to WT hearts slowed the rate of decline in ATP during ischemia to a level similar to that observed in Bcl-2 hearts, but addition of Oligo had no significant effect on the rate of decline in ATP in Bcl-2 hearts during ischemia. These data are consistent with Bcl-2-mediated inhibition of consumption of glycolytic ATP. Furthermore, mitochondria from Bcl-2 hearts have a reduced rate of consumption of ATP on uncoupler addition. This could be accomplished by limiting ATP entry into the mitochondria through the voltage-dependent anion channel, and/or the adenine nucleotide transporter, or by direct inhibition of the F1F0-ATPase. Immunoprecipitation showed greater interaction between Bcl-2 and voltage-dependent anion channel during ischemia. These data indicate that Bcl-2 modulation of metabolism contributes to cardioprotection. PMID- 15345652 TI - Tumor necrosis factor-alpha-converting enzyme (ADAM17) mediates GPIbalpha shedding from platelets in vitro and in vivo. AB - Interaction of the platelet receptor glycoprotein (GP) Ib-V-IX with von Willebrand factor exposed at a site of vascular injury is an essential step in the initiation of a hemostatic plug. Proteolytic cleavage (shedding) of the GPIbalpha subunit was first described >25 years ago, the protease mediating this event as well as its physiological function, however, have not been elucidated. We reported recently that shedding of GPIbalpha induced by platelet storage or mitochondrial injury involves a platelet-derived metalloproteinase(s). Here we show that GPIbalpha shedding in response to mitochondrial injury or physiological activation is inhibited in platelets obtained from chimeric mice, which express inactive tumor necrosis factor-alpha converting enzyme (TACE(DeltaZn/DeltaZn)) in blood cells only. Shedding was also inhibited in mouse and human platelets in the presence of 2 potent TACE inhibitors: TAP1 and TMI-1. Our data further suggest that TACE is important in the regulation of GPIbalpha expression in vivo because we observed an approximately 90% reduction in soluble GPIbalpha (glycocalicin) in plasma of TACE(DeltaZn/DeltaZn) chimeras as well as significantly increased levels of GPIbalpha on circulating platelets. In contrast, shedding of P-selectin from activated platelets was not affected by the mutation in TACE. Damaged TACE(DeltaZn/DeltaZn) platelets were further characterized by a markedly improved post-transfusion recovery and hemostatic function in mice. In conclusion, our data demonstrate that TACE is expressed in platelets and that it is the key enzyme mediating shedding of GPIbalpha. PMID- 15345653 TI - Flow-dependent regulation of endothelial Toll-like receptor 2 expression through inhibition of SP1 activity. AB - Innate immune system activation is associated with atherosclerotic lesion development. The specific sites of lesion development are believed to be defined by the shear stress of blood flow. Consequently, we investigated the responsiveness of human coronary artery endothelial cells (HCAECs) to Toll-like receptor (TLR) 2 and 4 agonists in an in vitro model of chronic laminar flow. HCAECs under chronic laminar flow were found to be normally responsive to lipopolysaccharide (and tumor necrosis factor) in terms of E-selectin expression but were found to be hyporesponsive to stimulation with the specific TLR2 ligands macrophage activating lipopeptide-2, PAM2-Cys, and Lip19; this was observed to be attributable to downregulation of TLR2 transcription and protein expression. We found that laminar flow induced SP1 serine phosphorylation by protein kinase CK2 and thereby blocked SP1 binding to the TLR2 promoter, which is required for TLR2 expression. This regulatory mechanism also blocked lipopolysaccharide- and tumor necrosis factor-induced TLR2 upregulation in HCAECs and could be important for suppression of other flow-sensitive endothelial proteins. These results extend the role of flow in controlling endothelial responsiveness. Given the current evidence that TLRs are proatherogenic, flow suppression of TLR2 expression may be atheroprotective. PMID- 15345654 TI - Mechanisms underlying conduction slowing and arrhythmogenesis in nonischemic dilated cardiomyopathy. AB - Heart Failure (HF) is associated with an increased risk of sudden death caused by ventricular tachyarrhythmias. Recent studies have implicated repolarization abnormalities and, in particular, exaggerated heterogeneity of transmural repolarization in the genesis of polymorphic ventricular tachycardia in a canine model of nonischemic dilated cardiomyopathy. The presence and degree to which conduction abnormalities play a role in arrhythmogenesis in this model are uncertain. HF was produced in dogs by rapid RV-pacing for 3 to 4 weeks. High resolution optical action potentials were recorded from epicardial and endocardial surfaces of arterially perfused canine wedge preparations isolated from LV and RV of normal and failing dogs. Cellular and molecular determinants of conduction were investigated using patch-clamp recordings, Western blot analysis, and immunocytochemistry. HF was associated with marked prolongation (by 33%) of the QRS duration of the volume conducted electrocardiogram and significant (>20%) slowing of epicardial and endocardial conduction velocities (CV) in both LV and RV. Cx43 expression was reduced by >40% in epicardial and endocardial layers of the LV, but was unchanged in the RV of failing hearts. Despite greater epicardial than endocardial Cx43 expression, epicardial CV was consistently slower (P<0.01). Immunocytochemical analysis revealed predominant colocalization of Cx43 with N cadherin in normal versus failing samples, because Cx43 was redistributed from the intercalated disk to lateral cell borders in failing tissue. Moreover, a significant (P<0.05) increase in hypophosphorylated Cx43 was detected in the LV and RV of failing hearts. Action potential upstroke velocities in isolated ventricular myocytes from normal and failing hearts were not different (P=0.8, not significant), and Masson trichrome staining revealed no significant change in fibrosis content in HF. Nonischemic dilated cardiomyopathy is associated with significant slowing of CV that was not directly related to reduced Cx43 expression. Changes in phosphorylation and localization of Cx43 may contribute to gap-junction dysfunction, CV slowing, and arrhythmias in HF. PMID- 15345655 TI - 17beta-estradiol reduces cardiomyocyte apoptosis in vivo and in vitro via activation of phospho-inositide-3 kinase/Akt signaling. AB - Female gender and estrogen-replacement therapy in postmenopausal women are associated with improved heart failure survival, and physiological replacement of 17beta-estradiol (E2) reduces infarct size and cardiomyocyte apoptosis in animal models of myocardial infarction (MI). Here, we characterize the molecular mechanisms of E2 effects on cardiomyocyte survival in vivo and in vitro. Ovariectomized female mice were treated with placebo or physiological E2 replacement, followed by coronary artery ligation (placebo-MI or E2-MI) or sham operation (sham) and hearts were harvested 6, 24, and 72 hours later. After MI, E2 replacement significantly increased activation of the prosurvival kinase, Akt, and decreased cardiomyocyte apoptosis assessed by terminal deoxynucleotidyltransferase dUTP nick-end labeling (TUNEL) staining and caspase 3 activation. In vitro, E2 at 1 or 10 nmol/L caused a rapid 2.7-fold increase in Akt phosphorylation and a decrease in apoptosis as measured by TUNEL staining, caspase 3 activation, and DNA laddering in cultured neonatal rat cardiomyocytes. The E2-mediated reduction in apoptosis was reversed by an estrogen receptor (ER) antagonist, ICI 182,780, and by phospho-inositide-3 kinase inhibitors, LY294002 and Wortmannin. Overexpression of a dominant negative-Akt construct also blocked E2-mediated reduction in cardiomyocyte apoptosis. These data show that E2 reduces cardiomyocyte apoptosis in vivo and in vitro by ER- and phospho-inositide-3 kinase-Akt-dependent pathways and support the relevance of these pathways in the observed estrogen-mediated reduction in myocardial injury. PMID- 15345656 TI - Passive stiffness changes caused by upregulation of compliant titin isoforms in human dilated cardiomyopathy hearts. AB - In the pathogenesis of dilated cardiomyopathy, cytoskeletal proteins play an important role. In this study, we analyzed titin expression in left ventricles of 19 control human donors and 9 severely diseased (nonischemic) dilated cardiomyopathy (DCM) transplant-patients, using gel-electrophoresis, immunoblotting, and quantitative RT-PCR. Both human-heart groups coexpressed smaller (approximately 3 MDa) N2B-isoform and longer (3.20 to 3.35 MDa) N2BA isoforms, but the average N2BA:N2B-protein ratio was shifted from approximately 30:70 in controls to 42:58 in DCM hearts, due mainly to increased expression of N2BA-isoforms >3.30 MDa. Titin per unit tissue was decreased in some DCM hearts. The titin-binding protein obscurin also underwent isoform-shifting in DCM. Quantitative RT-PCR revealed a 47% reduction in total-titin mRNA levels in DCM compared with control hearts, but no differences in N2B, all-N2BA, and individual N2BA transcripts. The reduction in total-titin transcripts followed from a decreased area occupied by myocytes and increased connective tissue in DCM hearts, as detected by histological analysis. Force measurements on isolated cardiomyofibrils showed that sarcomeric passive tension was reduced on average by 25% to 30% in DCM, a reduction readily predictable with a model of wormlike-chain titin elasticity. Passive-tension measurements on human-heart fiber bundles, before and after titin proteolysis, revealed a much-reduced relative contribution of titin to total passive stiffness in DCM. Results suggested that the titin isoform shift in DCM depresses the proportion of titin-based stiffness by approximately 10%. We conclude that a lower-than-normal proportion of titin-based stiffness in end-stage failing hearts results partly from loss of titin and increased fibrosis, partly from titin-isoform shift. The titin-isoform shift may be beneficial for myocardial diastolic function, but could impair the contractile performance in systole. PMID- 15345657 TI - Acetaminophen hepatotoxicity: NO to the rescue. AB - Severe liver injury as a result of overdose or chronic use of acetaminophen (paracetamol) remains a significant clinical problem, accounting for as much as 40% of cases of acute liver failure. The mechanisms underlying the liver injury caused by acetaminophen have become much better understood in recent years. In this issue, Fiorucci et al. report that delivery of nitric oxide (NO) in small amounts to the liver, via a novel derivative of the bile acid ursodeoxycholic acid, results in significant protection of the liver from acetaminophen-induced damage. NO appears to produce these beneficial actions through several mechanisms, including the suppression of synthesis of several proinflammatory cytokines. There is also substantial evidence that a NO-releasing derivative of acetaminophen offers several advantages over acetaminophen itself, including enhanced analgesic potency and reduced liver toxicity. PMID- 15345658 TI - Liver delivery of NO by NCX-1000 protects against acute liver failure and mitochondrial dysfunction induced by APAP in mice. AB - 1. NCX-1000, (3alpha, 5beta, 7beta)-3,7-dihydroxycholan-24oic acid[2-methoxy-4-[3 [4-(nitroxy)butoxy]-3-oxo-1-propenyl]phenyl ester, is a nitric oxide (NO) derivative of ursodeoxyxholic acid (UDCA) that selectively release NO in the liver. 2. Here, we demonstrated that administering mice with 40 micromol kg(-1) NCX-1000, but not UDCA, improves liver histopathology and reduces mortality caused by 330 micromol kg(-1) APAP from 60 to 25% (P<0.01). Administration of NCX 1000, in a therapeutic manner, that is, 2 h after acetaminophen (APAP) intoxication reduced mortality, improved liver histopathology and prevented liver IFN-gamma, TNF-alpha, Fas/Fas ligand and inducible nitric oxide synthase (iNOS) mRNA accumulation caused by APAP. 3. In vitro exposure of primary cultures of mouse hepatocytes to APAP, 6.6 mm, resulted in apoptosis followed by necrosis. Loss of cell viability correlates with early mitochondrial membrane potential (Deltapsi(m)) hyperpolarization followed by depolarization and cytochrome c translocation from mitochondria to cytosol. APAP-induced apoptosis associated with procaspase-3 and -9 cleavage, appearance of truncated Bid and activation of poly(ADP-ribose) polymerase (PARP). 4. Treating primary culture of hepatocytes with 5 microm cyclosporine and 10 microm trifluoperazine for eight resulted in significant reduction of apoptosis induced by APAP suggesting that loss of Deltapsim was mechanistically involved in apoptosis induced by APAP in vitro. 5. NCX-1000, but not UDCA, concentration-dependently (ED(50)=16 microm) protected against Deltapsi(m) depolarization and reduced transition from apoptosis to necrosis caused by 6.6 mm APAP. 6. Treating primary cultures of hepatocytes with the NO-donor DETA-NO, 100 microm, reduced apoptosis induced by APAP and prevented caspase activation. 7. In conclusion, NCX-1000 is effective in protecting against APAP-induced hepatotoxicity when administered in a therapeutic manner. This protection may involve the inhibition of apoptosis and the maintenance of mitochondrial integrity. PMID- 15345659 TI - P2X2 and P2Y1 immunofluorescence in rat neostriatal medium-spiny projection neurones and cholinergic interneurones is not linked to respective purinergic receptor function. AB - 1. The presence of ionotropic P2X receptors, targets of ATP in fast synaptic transmission, as well as metabotropic P2Y receptors, known to activate K(+) currents in cultured neostriatal neurones, was investigated in medium-spiny neurones and cholinergic interneurones contained in neostriatal brain slices from 5-26-day-old rats. 2. In these cells, adenosine-5'-triphosphate (ATP) (100-1000 microm), 2-methylthioadenosine-5'-triphosphate (2MeSATP), alpha,beta methyleneadenosine-5'-triphosphate (alpha,betameATP, 30-300 microm, each) and adenosine-5'-O-(3-thiotriphosphate (ATPgammaS) (100 microm) failed to evoke P2X receptor currents even when 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 0.1 microm), apyrase (10 U ml(-1)) or intracellular Cs(+) was used to prevent occluding effects of the ATP breakdown product adenosine, desensitisation of P2X receptors by endogenous ATP and an interference with the activation of K(+) channels, respectively. P2X receptor agonists were also ineffective in outside out patches withdrawn from the brain slice tissue. Muscimol (10 microm) evoked GABA(A) receptor-mediated currents under all these conditions. 3. When used as a control, locus coeruleus neurones responded with P2X receptor-mediated currents to ATP (300 microm), 2MeSATP and alpha,betameATP (100 microm, each). 4. ATP and adenosine-5'-diphosphate (ADP) (100 microm, each) did not activate K(+) currents in the neostriatal neurones. 5. Despite the observed lack of function, P2X(2) and P2Y(1) immunofluorescence was found in roughly 50% of the medium-spiny neurones and cholinergic interneurones. 6. A role of ATP in synaptic transmission to striatal medium-spiny neurones and cholinergic interneurones appears unlikely, however, the otherwise silent P2X and P2Y receptors may gain functionality under certain yet unknown conditions. PMID- 15345660 TI - Mechanism of the drug interaction between valproic acid and carbapenem antibiotics in monkeys and rats. AB - The Ministry of Health and Welfare, Japan banned coadministration of carbapenems, such as panipenem/betamipron (PAPM), meropenem (MEPM), and valproic acid (VPA) because clinical reports have indicated that the coadministration caused seizures in epileptic patients due to lowered plasma levels of VPA. In this study, we have clarified the mechanism of the drug-drug interaction using PAPM, MEPM, and doripenem [S-4661; (+)-(4R,5S,6S)-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-3 [[(3S,5S)-5-[(sulfamoylamino)methyl]-3-pyrrolidinyl]thio]-1-azabicyclo[3.2.0]hept 2-ene-2-caboxylic acid monohydrate], a newly synthesized carbapenem. In vitro experiments using monkey liver slices suggested that the apparent synthetic rate of VPA glucuronide (VPA-G) increased in the presence of carbapenems. However, no such increase was observed in the experiment using monkey liver microsomes. Although no increase of uridine 5'-diphosphate D-glucuronic acid was found in monkey liver slices in the presence of carbapenems, potent inhibitory activity of carbapenems for the hydrolysis of VPA-G was found in monkey and rat liver homogenate. In vivo hydrolysis of VPA-G was clearly shown by the existence of VPA in plasma after dosing of VPA-G to rats, and its inhibition by carbapenems was also clearly shown by the negligible levels of VPA in rat plasma after coadministration of carbapenems and VPA-G. These results clearly indicate one of the important causes of drug interaction as follows: carbapenems would inhibit the hydrolytic enzyme, which is involved in the hydrolysis of VPA-G to VPA, resulting in a decrease of plasma concentration of VPA. PMID- 15345661 TI - Paraoxonase and superoxide dismutase gene polymorphisms and noise-induced hearing loss. AB - BACKGROUND: Noise-induced cochlear epithelium damage can cause hearing loss in industrial workers. In experimental systems, noise induces the release of free radicals and may damage the cochlear sensorial epithelium. Therefore, genes involved in regulating the reactive oxygen species manganese-superoxide dismutase (SOD2) and the antioxidant paraoxonase (PON) could influence cochlea vulnerability to noise. We evaluated whether susceptibility to noise-induced hearing loss (NIHL) is associated with SOD2, PON1, and PON2 polymorphisms in workers exposed to prolonged loud noise. METHODS: We enrolled 94 male workers from an aircraft factory in the study. The SOD2 gene was screened by denaturing reversed-phase HPLC, and the PON1 (Q192R and M55L) and PON2 (S311C) polymorphisms were analyzed by PCR amplification followed by digestion with restriction endonucleases. RESULTS: Three known (A16V, IVS3-23T/G, and IVS3-60T/G) and two new SOD2 polymorphisms (IVS1+ 8A/G and IVS3+107T/A) were identified. Regression analysis showed that PON2 (SC+CC) [odds ratio (OR) = 5.01; 95% confidence interval (CI), 1.11-22.54], SOD2 IVS3-23T/G and IVS3-60T/G (OR = 5.09; 95% CI, 1.27-20.47), age (OR = 1.22; 95% CI, 1.09-1.36), and smoking (OR = 49.49; 95% CI, 5.09-480.66) were associated with NIHL. No association was detected for PON1 (QQ+RR) and PON1 (LL) genotypes. CONCLUSIONS: Our data suggest that SOD2 and PON2 polymorphisms, by exerting variable local tissue antioxidant roles, could predispose to NIHL. However, caution should be exercised in interpreting these data given the small sample size and the difficulty in matching cases to controls regarding the overwhelming risk factor, i.e., smoking at least 10 cigarettes/day. PMID- 15345662 TI - Specific immunoassays for detection of intact and cleaved forms of the urokinase receptor. AB - BACKGROUND: The cell surface receptor (uPAR) for urokinase plasminogen activator (uPA) is a strong prognostic marker in several types of cancer. uPA cleaves the three-domain protein uPAR(I-III) into two fragments: uPAR(I), which contains domain I; and uPAR(II-III), which contains domains II and III. Established immunoassays measure a combination of uPAR forms. Our aim was to design immunoassays for specific quantification of the individual forms of uPAR. METHODS: Using appropriate combinations of epitope-mapped monoclonal antibodies (Mabs) for capture and europium-labeled detection Mabs, we designed two-site sandwich time-resolved fluorescence immunoassays (TR-FIAs): TR-FIA 1 to measure uPAR(I-III) alone; TR-FIA 2 to measure both uPAR(I-III) and uPAR(II-III); and TR FIA 3 to measure uPAR(I). To avoid detection of uPAR(I-III) in TR-FIA 3, we used a combination of the peptide uPAR antagonist AE120 and a domain I antibody, R3. AE120 blocks the binding of R3 to uPAR(I-III). In contrast, AE120 does not interact with liberated domain I and therefore does not interfere with the binding of R3 to uPAR(I). RESULTS: The limits of quantification (CV <20%) determined by adding the proteins to uPAR-depleted plasma were <3 pmol/L in all three assays. The interassay CVs in plasma with added analytes were <11%, and recoveries were between 93% and 105%. Cross-reactivities of purified proteins in the three TR-FIAs were no more than 4%. Studies on chymotrypsin cleavage of uPAR and size-exclusion chromatography of plasma with and without added protein further supported the specificity of the assays. CONCLUSIONS: The three novel TR FIAs accurately quantify uPAR(I-III) alone, uPAR(I-III) together with uPAR(II III), and uPAR(I), respectively, in biological samples, including plasma, and thus are well suited for studies of the diagnostic and prognostic value of individual uPAR forms in cancer patients. PMID- 15345663 TI - Detection of antinuclear antibodies by solid-phase immunoassays and immunofluorescence analysis. AB - BACKGROUND: Antinuclear antibodies (ANAs) are associated with several inflammatory rheumatic diseases. The aim of the present work was to evaluate enzyme immunoassays (EIAs) and compare them with classic immunofluorescent analysis (IFA) for the detection of ANA. METHODS: Seven enzyme immunoassays were used in this study. All assays were applied as described by the manufacturers. Three populations were included in the study: (a) a population of patients with well-established autoimmune inflammatory disease (n = 102); (b) a population in which a rheumatic disease was diagnosed up to 5 years after an IFA was performed (n = 164); and (c) a population of consecutive outpatients suspected to have a rheumatic disease (n = 101). The current clinical diagnoses of the patients served as the standard against which performance of the assays was evaluated. RESULTS: In patients with well-established rheumatic disorders, the newly developed EIA in which HEp-2 extracts were included had sensitivities and specificities comparable to or in some instances better than the IFA. The assays without HEp-2 extracts included had significantly lower sensitivities and specificities. In the outpatient population, up to 51% of patients had positive ANA tests that did not correspond to classic ANA-associated disease. However, in the assays in which the HEp-2 extracts were not included, the false-positive rate was <10%. The false-negative rate judged against IFA differed from assay to assay and disease to disease and was mostly <10%. CONCLUSIONS: In this study, the sensitivities of EIAs and IFA were largely comparable. However, EIAs without HEp 2 extracts included had a low sensitivity but a high specificity, particularly in nonselected populations. The choice of test is highly dependent on the clinical setting in which the ANA test is to be used and on laboratory policy. PMID- 15345664 TI - High intraindividual variation of B-type natriuretic peptide (BNP) and amino terminal proBNP in patients with stable chronic heart failure. AB - BACKGROUND: Plasma B-type natriuretic peptide (BNP) and N-terminal proBNP (NT proBNP) are promising markers for heart failure diagnosis, prognosis, and treatment. Insufficient data on the intraindividual biological variation (CV(i)) of BNP and NT-proBNP hamper interpretation of changes in concentration on disease progression or treatment optimization. We therefore investigated CV(i) values in stable heart failure patients. METHODS: We recruited 43 patients with stable chronic heart failure living in Curacao (22 males, 21 females; median age, 63 years; range, 20-86 years; New York Heart Association classes I-III). Samples were collected for within-day CV(i) (n = 6; every 2 h starting at 0800), day-to day CV(i) (n = 5; samples collected between 0800 and 1000 on 5 consecutive days), and week-to-week CV(i) (n = 6; samples collected between 0800 and 1000 on the same day of the week for 6 consecutive weeks). NT-proBNP (Roche) and BNP (Abbott) were measured by immunoassay. RESULTS: Median (range) concentrations were 134 (0 1630) ng/L (BNP) and 570 (17-5048) ng/L (NT-proBNP). Analytical variation, week to-week CV(i), and reference change values were 8.4%, 40%, and 113% (BNP), and 3.0%, 35%, and 98% (NT-proBNP). Week-to week CV(i)s were inversely related to median BNP concentrations. Week-to week CV(i)s for BNP were 44% (BNP < or =350 ng/L) and 30% (BNP >350 ng/L). Both BNP and NT-proBNP increased between 0800 and 1000. Median NT-proBNP/BNP ratios were inversely related to median BNP concentrations. CONCLUSIONS: The high CV(i)s hamper interpretation of changes in BNP and NT-proBNP concentrations and may partly explain their poor diagnostic values in chronic heart failure. Easily modifiable determinants to lower CV(i) have not been identified. The value of BNP and NT-proBNP for chronic heart failure diagnosis, and especially for follow-up and treatment optimization of individuals, remains largely to be established. PMID- 15345665 TI - Beating ischemia: a new feat of EETs? PMID- 15345666 TI - Cell-matrix signaling and thrombospondin: another link to myocardial matrix remodeling. PMID- 15345667 TI - Influence of mechanical, cellular, and molecular factors on collateral artery growth (arteriogenesis). AB - Growth of collateral blood vessels (arteriogenesis) is potentially able to preserve structure and function of limbs and organs after occlusion of a major artery. The success of the remodeling process depends on the following conditions: (1) existence of an arteriolar network that connects the preocclusive with the postocclusive microcirculation; (2) activation of the arteriolar endothelium by elevated fluid shear stress; (3) invasion (but not incorporation) of bone marrow-derived cells; and (4) proliferation of endothelial and smooth muscle cells. Most organs of most mammals including man can rely on the existence of interconnecting arterioles in most organs and tissues with heart being the exception in rodents and pigs. Arterial occlusion lowers the pressure in the distal vasculature thereby creating a pressure gradient favoring increased flow through preexisting collaterals. This increases fluid shear stress leading to endothelial activation with cellular edema, upregulation of adhesion molecules, mitogenic-, thrombogenic-, and fibrinolytic factors, leading to monocyte invasion with matrix digestion. Smooth muscle cells migrate and proliferate and the vessel enlarges under the influence of increasing circumferential wall stress. Growth factors involved belong to the FGF family and signaling proceeds via the Ras/Raf- and the Rho cascades. Increases in vascular radius and wall thickness restore fluid shear stress and circumferential wall stress to normal levels and growth stops. Although increases in collateral vessel size are very substantial their maximal conductance amounts to only 40% of normal. Forced increases in FSS can reach almost 100%. PMID- 15345669 TI - Modeling alveolar capillary dysplasia. PMID- 15345668 TI - Heart valve development: endothelial cell signaling and differentiation. AB - During the past decade, single gene disruption in mice and large-scale mutagenesis screens in zebrafish have elucidated many fundamental genetic pathways that govern early heart patterning and differentiation. Specifically, a number of genes have been revealed serendipitously to play important and selective roles in cardiac valve development. These initially surprising results have now converged on a finite number of signaling pathways that regulate endothelial proliferation and differentiation in developing and postnatal heart valves. This review highlights the roles of the most well-established ligands and signaling pathways, including VEGF, NFATc1, Notch, Wnt/beta-catenin, BMP/TGF beta, ErbB, and NF1/Ras. Based on the interactions among and relative timing of these pathways, a signaling network model for heart valve development is proposed. PMID- 15345670 TI - Insulin-like growth factor-i regulation of hepatic scavenger receptor class BI. AB - High-density lipoprotein mediates a normal physiological process called reverse cholesterol transport. This process enables the transfer of cholesterol from peripheral tissues to the liver for further metabolism and eventual secretion in the form of bile. The scavenger receptor of the B class (SR-BI), human homolog of SR-BI, and CD36 and LIMPII analogous-1 (CLA-1) are different names for the same receptor that facilitates hepatocellular uptake of cholesterol from high-density lipoprotein. The pivotal role of this receptor in enterohepatic circulation of cholesterol and bile salts underlies our interest to study the regulation of hepatic SR-BI gene in response to the actions of IGF-I. The results of our studies showed that endogenous expression of SR-BI/CLA-1 was suppressed by exposure to GH or IGF-I in cultured HepG2 cells. This observation extended to a whole animal model of rats continuously infused with IGF-I. IGF-I decreased transcriptional activity of the SR-BI promoter. However, the inhibitory effect of IGF-I on SR-BI/CLA-1 promoter activity was abrogated by wortmannin, a specific inhibitor of phosphoinositide 3-kinase (PI3-K). Exposure of HepG2 cells to IGF-I elicited a rapid phosphorylation of Akt. We also demonstrated that the constitutively active form of both p110, a subunit of PI3-K, and Akt inhibited activity of the human SR-BI/CLA-1 promoter. Furthermore, the dominant-negative mutant of Akt abolished the ability of IGF-I to suppress activity of the SR BI/CLA-1 promoter. In conclusion, PI3-K/Akt pathways participate in IGF-I suppression of SR-BI/CLA-1 expression, which suggests that the activation of Akt plays an important role in cholesterol metabolism in liver. PMID- 15345671 TI - Connective tissue growth factor mediates high glucose effects on matrix degradation through tissue inhibitor of matrix metalloproteinase type 1: implications for diabetic nephropathy. AB - High glucose concentration inhibits matrix degradation and affects the activities of the enzymes responsible, the matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). Connective tissue growth factor (CTGF) expression is increased in diabetic nephropathy and is a downstream mediator of TGF-beta actions. However, whether CTGF regulates matrix degradation and the mechanism of effect in diabetes has not been reported. Human mesangial cells were cultured in media containing 5 or 25 mM glucose and, in some experiments, with recombinant human (rh)CTGF (0-1000 ng/ml) and/or appropriate neutralizing antibodies. Matrix degradation was inhibited by rhCTGF in a dose-dependent manner, and the decrease in matrix degradation caused by high glucose and by TGF-beta was significantly attenuated by addition of CTGF-neutralizing antibody (by 40.2 and 69.1%, respectively). Similar to 25 mM glucose, addition of rhCTGF increased MMP-2, TIMP 1, and TIMP-3 mRNA by 2.5-, 2.1-, and 1.6-fold, respectively (P < 0.05) but had no effect on membrane-type (MT)1-MMP or TIMP-2. Addition of TIMP-1 antibody to conditioned medium abolished the decrease in degradation caused by rhCTGF and partially prevented (by 79%) the glucose-induced inhibition of matrix degradation. In vivo studies of glomeruli from diabetic and control rats showed that intensive insulin treatment prevented the increase in expression of CTGF and TIMP-1 and attenuated the decreased matrix degradation seen in diabetes. In summary, CTGF inhibits matrix degradation by increasing TIMP-1 expression, and by this action it contributes to the inhibition of matrix breakdown by high glucose, implying that CTGF has a role in the reduced matrix degradation observed in diabetic nephropathy. PMID- 15345672 TI - Estradiol regulates the thioredoxin antioxidant system in the mouse uterus. AB - The rodent uterus responds to acute estradiol (E2) treatment with a series of well characterized physiological responses. In a recent screen for genes involved in this response, we found that several genes in the thioredoxin (Txn) pathway were rapidly modified after E2 treatment in the mouse uterus. Txn is a 12-kDa protein with multiple roles in the cell, including protection against oxidative stress and apoptosis, regulation of transcription factor activity, and regulation of cellular proliferation. Txn in combination with Txn reductase (Txnrd) and Txn interacting protein (Txnip) constitute the mammalian Txn pathway. This pathway exists in multiple locations in the cell, including the cytosol and mitochondria. To analyze the levels of Txn, Txnrd, and Txnip in the uterus, we treated ovariectomized adult mice with a time course of E2 and analyzed mRNA levels by real-time PCR. E2 rapidly decreased the expression of Txnip, but increased the levels of cytosolic Txn1 and Txnrd1 as well as mitochondrial Txn2. Using the ER antagonist, ICI 182,780, and mice lacking functional estrogen receptor alpha (ERalpha), we demonstrate that these E2-mediated changes require ERalpha, but not ERbeta. The repression of Txnip by E2 was also demonstrated in vitro in MCF-7 human breast cancer cells. This repression was blocked by treatment with the histone deacetylase inhibitor, trichostatin A, suggesting that repression by E2 may involve regulation of histone acetylation. We conclude that the rapid E2 mediated activation of the Txn pathway is an important step in the response of the mammalian uterus to estrogen. PMID- 15345673 TI - Ataxia-telangiectasia mutated gene controls insulin-like growth factor I receptor gene expression in a deoxyribonucleic acid damage response pathway via mechanisms involving zinc-finger transcription factors Sp1 and WT1. AB - The IGF-I receptor (IGF-IR) has a central role in cell cycle progression as well as in the establishment of the transformed phenotype. Increased expression of the IGF-IR gene, in addition, is correlated with acquisition of radioresistance for cell killing. The ataxia-telangiectasia mutated (ATM) gene product has a pivotal role in coordinating the cellular response to DNA damage. The present study was aimed at testing the hypothesis that the ability of ATM to coordinate the DNA damage response that will lead to cell survival or, alternatively, to apoptosis depends, to a significant extent, on its capacity to control IGF-IR gene expression. The potential involvement of ATM in regulation of IGF-IR expression and function was investigated in isogenic cells with and without ATM function [AT22IJE-T/pEBS7 (ATM -/-) and ATM-corrected AT22IJE-T/YZ5 (ATM +/+) cells and 293 human embryonic kidney cells transfected with small interfering RNAs targeted to ATM]. In addition, the effect of ATM on IGF-IR expression was assessed in nonisogenic cells with ATM function (HFF + human telomerase reverse transcriptase) and without ATM function (GM5823 + human telomerase reverse transcriptase). Results obtained showed that IGF-IR gene expression and IGF-IR promoter activity were largely reduced in ATM -/- cells. Addition of the radiomimetic agent neocarzinostatin for 4 h, however, induced a significant increase in IGF-IR levels in cells without ATM function. In addition, IGF-I induced IGF-IR and insulin receptor substrate-1 phosphorylation were greatly impaired in ATM-deficient cells. Furthermore, we identified zinc-finger transcription factors Sp1 and WT1 as potential mediators of the effect of ATM on IGF-IR gene expression. The present data suggests that the IGF-IR gene is a novel downstream target in an ATM-mediated DNA damage response pathway. Deregulated expression of the IGF-IR gene after ionizing radiation may be linked to genomic instability and enhanced transforming capacity. PMID- 15345674 TI - Expression and regulation of type 2 iodothyronine deiodinase in rat aorta media. AB - Here, we have found that type 2 iodothyronine deiodinase (D2) is present in rat aorta media and that there is a circadian variation in the D2 expression. The D2 mRNA was approximately 4-fold higher at 0900 h than at 2100 h, and the activity was approximately 6-fold higher at noon than at 2100 h. The increase in aorta media D2 activity is preceded by the increase in its mRNA. The increase in D2 mRNA and activity in the circadian variation was reduced by the administration of prazosin, an alpha1-adrenergic antagonist, and propranolol, a beta- adrenergic antagonist. Furthermore, phenylephrine, an alpha1-adrenergic agonist, and isoproterenol, a beta-adrenergic agonist, caused a significant increase in D2 mRNA and activity. In the hypothyroid rats, aorta mediae D2 mRNA at both 0900 and 2100 h were not significantly different when compared with those in the euthyroid rats. On the other hand, aorta mediae D2 activity at both 1200 and 2100 h in the hypothyroid rats were approximately 2-fold higher. From these results, we suggest that D2 activity of rat aorta media is increased by both alpha1- and beta adrenergic stimulation, at least partly, at the pretranslational level. We also suggest that both alpha1- and beta-adrenergic mechanisms may be involved, at least partly, in the circadian variation of the activity. In the hypothyroid state, the aorta media D2 activity is increased mainly by the posttranslational mechanism, and the similar circadian variation of the D2 expression is present as in the euthyroid state. PMID- 15345675 TI - Thyrotropin-releasing hormone-stimulated thyrotropin expression involves islet brain-1/c-Jun N-terminal kinase interacting protein-1. AB - Islet-brain-1 (IB1)/c-Jun N-terminal kinase interacting protein 1 (JIP-1) is a scaffold protein that is expressed at high levels in neurons and the endocrine pancreas. IB1/JIP-1 interacts with the c-Jun N-terminal kinase and mediates the specific physiological stimuli (such as cytokines). However, the potential role of the protein in the pituitary has not been evaluated. Herein, we examined expression of the gene encoding IB1/JIP-1 and its translated product in the anterior pituitary gland and a pituitary cell line, GH3. We then examined the potential role of IB1/JIP-1 in controlling TSH-beta gene expression. Exposure of GH3 cells to TRH stimulated the expression of IB1/JIP-1 protein levels, mRNA, and transcription of the promoter. The increase of IB1/JIP-1 content by transient transfection study of a vector encoding IB1/JIP-1 or by the stimulation of TRH stimulates TSH-beta promoter activity. This effect is not found in the presence of a mutated nonfunctional (IB1S59N) IB1/JIP-1 protein. Together, these facts point to a central role of the IB1/JIP-1 protein in the control of TRH-mediated TSH-beta stimulation. PMID- 15345676 TI - Peroxisome proliferator-activated receptor gamma-dependent activation of p21 in Panc-28 pancreatic cancer cells involves Sp1 and Sp4 proteins. AB - 1,1-Bis(3'-indolyl)-1-(p-trifluoromethylphenyl)methane (DIM-C-pPhCF3) and troglitazone activate peroxisome proliferator-activated receptor gamma (PPARgamma) in Panc-28 pancreatic cancer cells and also inhibit cell proliferation. DIM-C-pPhCF3 was more active than troglitazone and was used as a model to investigate the mechanism of PPARgamma-dependent inhibition of Panc-28 cell growth. DIM-C-pPhCF3 significantly inhibited G0/G1-->S phase progression, as determined by FACS analysis, and this was associated with decreased retinoblastoma protein phosphorylation and increased p21 protein and mRNA expression, but no change in p27 or cyclin D1. PPARgamma antagonists blocked DIM C-pPhCF3-induced growth inhibition and induction of p21 protein, and similar inhibitory effects were observed in Panc-28 cells transfected with a construct (pWWP) containing a -2325 to +8 p21 promoter insert. Deletion analysis of the p21 promoter indicated that PPARgamma-dependent activation of p21 promoter constructs by DIM-C-pPhCF3 required GC-rich sites 3 and 4 in the proximal region (-124 to 60) of the p21 promoter. The results of RNA interference and protein expression/DNA binding assays suggest that DIM-C-pPhCF3 induced p21 expression through a novel mechanism that involves PPARgamma interactions with both Sp1 and Sp4 proteins bound to the proximal GC-rich region of the p21 promoter. PMID- 15345677 TI - Developmental expression of estrogen receptor (ER) alpha and ERbeta in the hamster ovary: regulation by follicle-stimulating hormone. AB - Perinatal expression of estrogen receptor (ER) protein and mRNA and the influence of FSH on this process were examined by immunofluorescence and RT-PCR using ovaries from fetal (d 13-15 of gestation) and postnatal [postnatal d 1-15 (P1 P15)] hamsters and from 8-d-old hamsters exposed in utero to an anti-FSH serum on d 12 of gestation and saline or equine chorionic gonadotropin (eCG) on P1. A few somatic cells expressing ERalpha immunoreactivity appeared first on d 14 of gestation and increased markedly by P8-P15 in the interstitial cells and granulosa cells of primordial follicles. In contrast, appreciable ERbeta immunoreactivity was localized on d 13 of gestation, and more cells expressed ERbeta immunoreactivity by P1-P8. By P7, ERbeta immunoreactivity was present in cells adjacent to the oocytes, and by P8, ERbeta was preferentially localized in the granulosa cells. Receptor immunoreactivities decreased markedly in P8 ovaries exposed in utero to the FSH antiserum but were reversed with postnatal eCG replacement. Oocytes and somatic cells expressed ERalpha and ERbeta mRNA, and levels of ER mRNA in the ovary increased by P7-P8, corresponding to the appearance of primordial follicles. Thereafter, only ERbeta mRNA levels increased progressively with postnatal ovary development. Similar to ER protein, mRNA levels decreased significantly in FSH antiserum-treated ovaries but were restored by eCG. These results indicate that both ER subtypes are expressed in undifferentiated somatic cells and the oocytes during perinatal ovary development in the hamster; however, ERbeta expression segregates with the differentiation of granulosa cells. Furthermore, ER expression and differentiation of somatic cells to granulosa cells depend on perinatal FSH action. PMID- 15345678 TI - Rapid nongenomic effects of 3,5,3'-triiodo-L-thyronine on the intracellular pH of L-6 myoblasts are mediated by intracellular calcium mobilization and kinase pathways. AB - L-T3 and L-T4 activated the Na+/H+ exchanger of L-6 myoblasts, with a fast nongenomic mechanism, both in the steady state and when cells undergo acid loading with ammonium chloride. Monitored with the intracellular pH-sensitive fluorescent probe 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein, activation of the exchanger appeared to be initiated at the plasma membrane, because T3-agarose reproduced the effect of L-T3, and triiodothyroacetic acid, a hormone analog previously shown to inhibit membrane actions of thyroid hormone, blocked the action of L-T3 on the exchanger. We show here for the first time that transduction of the hormone signal in this nongenomic response requires tyrosine kinase-dependent phospholipase C activation and two different signaling pathways: 1) mobilization of intracellular calcium, assessed by the fluorescent probe fura 2, through activation of inositol trisphosphate receptors and without contributions from extracellular calcium or ryanodine receptors; and 2) protein phosphorylation involving protein kinase C and MAPK (ERK1/2), as shown by the use of kinase inhibitors and by immunoblotting for activated kinases. PMID- 15345679 TI - Expression of human hormone-sensitive lipase (HSL) in postmeiotic germ cells confers normal fertility to HSL-deficient mice. AB - Hormone-sensitive lipase (HSL, Lipe, E.C.3.1.1.3) is a multifunctional fatty acyl esterase that is essential for male fertility and spermatogenesis and that also plays important roles in the function of adipocytes, pancreatic beta-cells, and adrenal cortical cells. Gene-targeted HSL-deficient (HSL-/-) male mice are infertile, have a 2-fold reduction in testicular mass, a 2-fold elevation of the ratio of esterified to free cholesterol in testis, and unique morphological abnormalities in round and elongating spermatids. Postmeiotic germ cells in the testis express a specific HSL isoform. We created transgenic mice expressing a normal human testicular HSL cDNA from the mouse protamine-1 promoter, which mediates expression specifically in postmeiotic germ cells. Testicular cholesteryl esterase activity was undetectable in HSL-/- mice, but in HSL-/- males expressing the testicular transgene, activity was 2-fold greater than normal. HSL transgene mRNA became detectable in testes between 19 and 25 days of age, coinciding with the first wave of postmeiotic transcription in round spermatids. In contrast to nontransgenic HSL-/- mice, HSL-/- males expressing the testicular transgene were normal with respect to fertility, testicular mass, testicular esterified/free cholesterol ratio, and testicular histology. Their cauda epididymides contained abundant, normal-appearing spermatozoa. We conclude that human testicular HSL is functional in mouse testis and that the mechanism of infertility in HSL-deficient males is cell autonomous and resides in postmeiotic germ cells, because HSL expression in these cells is in itself sufficient to restore normal fertility. PMID- 15345681 TI - Transcriptional regulation of agouti-related protein (Agrp) in transgenic mice. AB - Agouti-related protein (Agrp) encodes a hypothalamic neuropeptide that promotes positive energy balance by stimulating food intake and reducing energy expenditure. Agrp expression in the brain is restricted to neurons within the arcuate nucleus of the hypothalamus, and expression levels are elevated as a consequence of food deprivation. We tested a series of bacterial artificial chromosome reporter constructs with varying amounts of sequence flanking the Agrp transcription unit in transgenic mice to identify and refine a region of DNA capable of recapitulating characteristics of Agrp expression. We report that a 42.5-kb region upstream of Agrp, containing three distinct regions that are evolutionarily conserved between mouse and human, is necessary and sufficient to consistently drive reporter expression specifically within AgRP neurons in a fasting-responsive manner. In addition, we demonstrate that this region allows for the stable expression of Cre recombinase in transgenic mice, providing a genetic tool for studying anabolic neural circuits that control energy balance. PMID- 15345680 TI - Postnatal regression of hypothalamic dopaminergic neurons in prolactin-deficient Snell dwarf mice. AB - Both Snell (Pit-1(dw) or (dwj), dw/dw) and Ames (Prophet of Pit-1(df), df/df) dwarf mice fail to produce prolactin (PRL) as well as GH due to deficient transcription factor Pit-1 activity and have reduced numbers of hypothalamic PRL inhibiting area A12 tuberoinfundibular dopaminergic (TIDA) neurons. It has been reported that the TIDA deficit in Ames dwarf mice develops postnatally as a reduction in number after an initial increase that is comparable to that of normal siblings. The present study was designed to characterize A12 TIDA neuronal development in the Snell dwarf (dw/dw) compared with littermate normal mice. Brains of normal (DW/?) and dw(j)/dw(j) mice were examined at 7, 14, 21, 30, and > or = 60 postnatal days (d) by catecholamine fluorescence and quantification of neuron number after tyrosine hydroxylase immunostaining in dopaminergic (DA) areas A12, A13 (medial zona incerta), and A14 (periventricular nucleus). Fluorescence was less in dw/dw than in DW/? A12 perikarya and median eminence but was not reduced in other DA areas, such as substantia nigra, at all ages; A12 fluorescence was virtually absent in Snell dwarf adults. Numbers of TIDA neurons were comparable in normal and Snell dwarf mice at 7 d. In normal (DW/?) mice, A12 neurons increased in number to adult levels at 14 d and were significantly higher than in Snell dwarf (dw/dw) mice at 14 d (P < 0.05) and at subsequent ages (P < 0.01). In Snell dwarf mice, numbers of A12 neurons did not differ at 7, 14, and 21 d, decreased at 30 d (P < 0.05), and reached, at 60 d, 23% of the population in normal sibling mice (P < 0.01 compared with earlier ages). Neuron numbers in nonhypophysiotropic DA area A13 did not vary with age or phenotype. In A14, cell number was higher in both phenotypes at 14 d (P < 0.05 for DW/?; P < 0.01 for dw/dw); neuron number was lower in dw/dw than in DW/? mice at 30 d (P < 0.05) and 60 d (P < 0.01). Thus, compared with normal mice of the same strain, the A12 deficit is more severe in Snell (dw/dw) than in Ames (df/df) dwarf hypothalamus (48% of DF/?), as previously reported, and develops as a decline from the population present at 7 d rather than first increasing. A reduction in A14 neuron number also occurs in the Snell dwarf. Treatment of DW/dw- and dw/dw-containing litters with ovine PRL (50 mug/d, ip), beginning at 12 or 7 d and continuing until 42 d, resulted in TIDA neuron numbers in Snell dwarfs that were lower than those in normal siblings (P < 0.01 for both) but were higher than in untreated adult dwarfs and comparable to the TIDA population size in dwarfs at 7 d, indicating that PRL maintained this maximal number and prevented TIDA neuron dedifferentiation, which occurs in dwarf postnatal development. PMID- 15345682 TI - Interaction of stress-activated protein kinase-interacting protein-1 with the interferon receptor subunit IFNAR2 in uterine endometrium. AB - During early pregnancy in ruminants, a type I interferon (IFN-tau) signals from the conceptus to the mother to ensure the functional survival of the corpus luteum. IFN-tau operates through binding to the type I IFN receptor (IFNR). Here we have explored the possibility that IFNAR2, one of the two subunits of the receptor, might interact with hitherto unknown signal transduction factors in the uterus that link IFN action to pathways other than the well established Janus kinase-signal transducer and activator of transcription pathways. A yeast two hybrid screen of an ovine (ov) endometrial cDNA library with the carboxyl terminal 185 amino acids of ovIFNAR2 as bait identified stress-activated protein kinase-interacting protein 1 (ovSin1) as a protein that bound constitutively through its own carboxyl terminus to the receptor. ovSin1 is a little studied, 522-amino acid-long polypeptide (molecular weight, 59,200) that is highly conserved across vertebrates, but has identifiable orthologs in Drosophila and yeast. It appears to be expressed ubiquitously in mammals, although in low abundance, in a wide range of mammalian tissues in addition to endometrium. Sin1 mRNA occurs in at least two alternatively spliced forms, the smaller of which lacks a 108-bp internal exon. ovSin1, although not exhibiting features of a membrane-spanning protein, such as IFNAR2, is concentrated predominantly in luminal and glandular epithelial cells of the uterine endometrium. When ovSin1 and ovIFNAR2 are coexpressed, the two proteins can be coimmunoprecipitated and colocalized to the plasma membrane and to perinuclear structures. Sin1 provides a possible link among type I IFN action, stress-activated signaling pathways, and control of prostaglandin production. PMID- 15345683 TI - Altering dietary nutrient intake that reduces glycogen content leads to phosphorylation of nuclear p38 MAP kinase in human skeletal muscle: association with IL-6 gene transcription during contraction. AB - To determine the effect of glycogen availability and contraction on intracellular signaling and IL-6 gene transcription, eight males performed 60 min of exercise on two occasions: either with prior ingestion of a normal (Con) or low carbohydrate (LCHO) diet that reduced pre-exercise muscle glycogen content. Muscle biopsies were obtained and analyzed for IL-6 mRNA. In addition, nuclear proteins were isolated from the samples and analyzed for the mitogen- activated protein kinases (MAPK) c-jun amino-terminal kinase (JNK) 1 and 2 and p38 MAPK. Nuclear fractions were also analyzed for the phosphorylated forms of JNK (p-JNK) and p38 MAPK (p-p38 MAPK) and the abundance of the nuclear transcription factors nuclear factor of activated T cells (NFAT) and nuclear factor kappa-beta (NF kappabeta). No differences were observed in the protein abundance of total JNK 1/2, p38 MAPK, NFAT, or NF-kappabeta before exercise, but the nuclear abundance of p-p38 MAPK was higher (P<0.05) in LCHO. Contraction resulted in an increase (P<0.05) in nuclear p-JNK 1/2, but there were no differences when comparing CON with LCHO. The fold increase in IL-6 mRNA with contraction was potentiated (P<0.05) in LCHO. A correlation between pre-exercise nuclear phosphorylated p38 MAPK and contraction-induced fold increase in IL-6 mRNA was performed, revealing a highly significant correlation (r=0.96; P<0.01). We next incubated L6 myotubes in ionomycin (a compound known to induce IL-6 mRNA) with or without the pyridinylimidazole p38 MAPK inhibitor SB203580. Treatments did not affect total nuclear p38 MAPK, but ionomycin increased (P<0.05) both nuclear p-p38 MAPK and IL 6 mRNA. The addition of SB203580 to ionomycin decreased (P<0.05) nuclear p-p38 MAPK and totally abolished (P<0.05) the ionomycin- induced increase in IL-6 mRNA. These data suggest that reduced carbohydrate intake that results in low intramuscular glycogen leads to phosphorylation of p38 MAPK at the nucleus. Furthermore, phosphorylation of p38 MAPK in the nucleus appears to be an upstream target for IL-6, providing new insights into the regulation of IL-6 gene transcription. PMID- 15345684 TI - INGAP peptide improves nerve function and enhances regeneration in streptozotocin induced diabetic C57BL/6 mice. AB - INGAP peptide comprises the core active sequence of Islet Neogenesis Associated Protein (INGAP), a pancreatic cytokine that can induce new islet formation and restore euglycemia in diabetic rodents. The ability of INGAP peptide in vitro to enhance nerve growth from sensory ganglia suggests its potential utility in peripheral nerve disorders. In this study, INGAP peptide was administered alone or in combination with insulin to streptozotocin-induced diabetic mice exhibiting signs of peripheral neuropathy. Following a 2-wk treatment period, thermal hypoalgesia in diabetic mice was significantly improved in groups that received INGAP peptide, without development of hyperalgesia. Explanted dorsal root ganglia (DRG) from these groups showed enhanced nerve outgrowth and evidence of increased mitochondrial activity. Western blotting experiments revealed attenuation of neurofilament hyperphosphorylation, up-regulation of beta-tubulin and actin, and increased phosphorylation of the transcription factor STAT3 in DRG. These findings suggest that INGAP peptide can activate some of the signaling pathways implicated in nerve regeneration in sensory ganglia, thereby providing a means of improvement of nociceptive dysfunction in the peripheral nervous system. PMID- 15345685 TI - Retinoid-modulated MAT1 ubiquitination and CAK activity. AB - Human cyclin-dependent kinase (CDK)-activating kinase (CAK) has a dual function in cross-regulation of cell cycle and differentiation, whereas menage a trois 1 (MAT1) assembles CAK and determines CAK's substrate specificity. Although the dynamic state of MAT1 protein levels is found to modulate CAK activity, how intracellular regulation of MAT1 controls CAK activity is unknown. Recent studies demonstrate that retinoic acid (RA)-induced human HL60 cell proliferation/differentiation (P/D) transition is accompanied by MAT1 degradation and decreased CAK phosphorylation of retinoic acid receptor alpha (RARa). Thus, we investigated the biochemical pathway of MAT1 degradation and its relationship with CAK phosphorylation of RARa. We find that RA induces ubiquitination proteolysis of MAT1 and that ubiquitin-proteasome targets CAK-free MAT1 only. RA induced MAT1 ubiquitination reduces CAK abundance and decreases CAK phosphorylation of RARalpha, whereas inhibition of MAT1 ubiquitination resists this RA-effect. These findings reveal that RA induces MAT1 ubiquitination to decrease CAK phosphorylation of RARalpha, suggesting a novel mechanism of RA mediated P/D transition in which MAT1 ubiquitination may act as an integral part of RA-effect to decrease CAK activity in the switch from proliferation to differentiation. PMID- 15345686 TI - A novel bifunctionality: PAT1 and PAT2 mediate electrogenic proton/amino acid and electroneutral proton/fatty acid symport. AB - Recently, the PAT family of proton-dependent amino acid transporters has been identified as a novel class of mammalian amino acid symporters. PAT1 and PAT2 members mediate electrogenic uptake of small, neutral amino acids and derivatives by cotransport of protons. Analysis of the structural requirements for substrate recognition by PAT1 identified that a free amino group in a substrate is not essential for recognition. We therefore hypothesized that PAT1 and its ortholog PAT2 may also be able to recognize and transport the homologous short-chain fatty acids (SCFAs) such as acetate, propionate, and butyrate. We examined in Xenopus laevis oocytes whether the SCFAs interact with the transporter by employing flux studies, electrophysiology and intracellular pH recordings. SCFAs did not induce positive inward currents but inhibited glycine-induced transport currents. PAT mediated uptake of radiolabeled proline was also dose-dependently reduced by SCFA and could be described by first order competition kinetics with apparent Ki values for butyrate of 6.0 +/- 0.7 and 7.6 +/- 1.3 mM for PAT1 and PAT2, respectively. Acetate as well as propionate uptake was significantly enhanced in oocytes expressing PAT1 or PAT2. An electroneutral H+/SCFA symport mode was demonstrated by recording intracellular pH changes under voltage clamp conditions with rate constants for the initial intracellular acidification in the presence of SCFAs significantly increased in PAT-expressing oocytes. In conclusion, our data demonstrate that the PAT1 and PAT2 proteins are capable to transport selected SCFAs in an electroneutral and the homologous amino acids in an electrogenic mode and are therefore a paradigm for bifunctional solute carriers. PMID- 15345687 TI - Endothelin induces differentiation of ANP-EGFP expressing embryonic stem cells towards a pacemaker phenotype. AB - Currently, only limited insight into mechanisms promoting the differentiation and specification of the mammalian cardiac conduction system is available. Therefore, we established a murine embryonic stem (ES) cell line stably expressing the enhanced green fluorescent protein (EGFP) under the transcriptional control of the human atrial natriuretic peptide (ANP) promoter to further characterize the development of very early stages of the mammalian cardiac conduction tissue. The cardiac nature of ANP-EGFP positive cells was confirmed by immunostaining. In ANP EGFP expressing ES cell-derived cardiomyocytes, a distinct sublineage of pacemaker cells could be identified. Pacemaker cells displayed a spindle shape and exhibited a higher spontaneous beating rate, faster If current activation and larger If current densities compared with triangular atrial-like cardiocytes. Exposure to endothelin-1 significantly increased the percentage of pacemaker-like cells without affecting their electrophysiological properties. These findings were corroborated by immunostaining with antibodies against connexin 40 and connexin 45, known markers for cardiac conduction tissue. Conversely, treatment of ANP-EGFP expressing ES cells with neuregulin-1 exhibited no effect on differentiation. These results indicate that ANP-EGFP expression enables the identification of ES cell-derived pacemaker cells by their fluorescence and morphology and that endothelin-1 promotes the development of ANP-EGFP positive cardiomyocytes to a pacemaker-like phenotype. PMID- 15345688 TI - Amyloidogenesis recapitulated in cell culture: a peptide inhibitor provides direct evidence for the role of heparan sulfate and suggests a new treatment strategy. AB - To date 22 different polypeptides, including Abeta in Alzheimer's disease and PrP(Sc) in prion disorders, are known to re-fold and assemble into highly organized fibrils, which associate with heparan sulfate (HS) proteoglycans to form tissue deposits called amyloid. Mononuclear phagocytes have long been thought to be involved in this process, and we describe a monocytic cell culture system that can transform the acute-phase protein serum amyloid A (SAA1.1) into AA-amyloid and appears to recapitulate all the main features of amyloidogenesis observed in vivo. These features in common include nucleation-dependent kinetics, identical proteolytic processing of SAA1.1, and co-deposition of HS with the fibrils. Heparin and polyvinylsulfonate previously reported to block AA amyloidogenesis in mice are also effective inhibitors in this cell culture model. Furthermore, a synthetic peptide (27-mer) corresponding to a HS binding site of SAA, blocks amyloid deposition at a concentration that is several-orders-of magnitude lower than any other peptide-based inhibitor previously reported. The 27-mer's inhibitory activity may target the amyloidogenic pathway specifically as it does not interfere with the binding of SAA to monocytes. These data provide direct evidence that SAA1.1:HS interactions are a critical step in AA amyloidogenesis and suggest a novel treatment strategy for other amyloidoses. PMID- 15345689 TI - Putting copper into action: copper-impregnated products with potent biocidal activities. AB - Copper ions, either alone or in copper complexes, have been used for centuries to disinfect liquids, solids, and human tissue. Today copper is used as a water purifier, algaecide, fungicide, nematocide, molluscicide, and antibacterial and antifouling agent. Copper also displays potent antiviral activity. We hypothesized that introducing copper into clothing, bedding, and other articles would provide them with biocidal properties. A durable platform technology has been developed that introduces copper into cotton fibers, latex, and other polymeric materials. This study demonstrates the broad-spectrum antimicrobial (antibacterial, antiviral, antifungal) and antimite activities of copper impregnated fibers and polyester products. This technology enabled the production of antiviral gloves and filters (which deactivate HIV-1 and other viruses), antibacterial self-sterilizing fabrics (which kill antibiotic-resistant bacteria, including methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci), antifungal socks (which alleviate symptoms of athlete's foot), and anti-dust mite mattress covers (which reduce mite-related allergies). These products did not have skin-sensitizing properties, as determined by guine pig maximization and rabbit skin irritation tests. Our study demonstrates the potential use of copper in new applications. These applications address medical issues of the greatest importance, such as viral transmissions; nosocomial, or healthcare-associated, infections; and the spread of antibiotic-resistant bacteria. PMID- 15345690 TI - CpG motifs in bacterial DNA delay apoptosis of neutrophil granulocytes. AB - Human neutrophil granulocytes die rapidly, and their survival is contingent upon rescue from programmed cell death by signals from the environment. We now show that a novel signal for delaying neutrophil apoptosis is unmethylated CpG motifs prevalent in bacterial DNA (CpG- DNA). Human neutrophils express toll-like receptor 9 that recognizes these motifs. CpG-DNA, but not mammalian DNA or methylated bacterial DNA, markedly enhanced neutrophil viability by delaying spontaneous apoptosis. Endosomal maturation of CpG-DNA is prerequisite for these actions and was coupled to concurrent activation of the extracellular signal regulated kinase (ERK) and phosphatidylinositol 3-kinase/Akt signaling pathways, leading to phosphorylation of BAD at Ser112 and Ser136, respectively, and to prevention of decreases in mitochondrial transmembrane potential, cytochrome c release and caspase-3 activation. Consistently, pharmacological inhibition of either ERK or phosphatidylinositol 3-kinase partially reversed these actions of CpG-DNA; however, they did not produce additive inhibition. Furthermore, intravenous injection of CpG-DNA (200 microg/kg) into rats evoked slight decreases in blood pressure and induced a modest leukocytosis, whereas it effectively suppressed neutrophil apoptosis as assayed ex vivo. Our results indicate that unmethylated CpG motifs in bacterial DNA promote neutrophil survival by suppressing the apoptotic machinery and may therefore contribute to prolongation and amplification of inflammation. PMID- 15345691 TI - Protective effect of TAT-delivered alpha-synuclein: relevance of the C-terminal domain and involvement of HSP70. AB - Alpha-synuclein (alpha-syn) is a 140-amino acid presinaptic protein whose mutations A30P and A53T have been linked to familiar Parkinson's disease (PD). Many data suggest that alpha-syn aggregation is the key event that triggers alpha syn-mediated neurotoxicity. Nevertheless, other lines of evidence proposed a protective role of alpha-syn against oxidative stress (a major feature of PD), even if the exact mechanism of this protective action and the role of the pathogenetic mutations to this respect have not been elucidated yet. To address these points, we developed an in vitro model of oxidative stress by exposing PC12 cells to hydrogen peroxide (H2O2) (150 microM) for 72 h, and we evaluated alpha syn-mediated protection delivering increasing amounts of alpha-syn (wild type [WT] or mutated) inside cells using the fusion proteins TAT-alpha-syn (WT, A30P, and A53T). We found that nanomolar amounts of TAT-alpha-syn-mediated protected against oxidative stress and other cellular injuries (6-hydroxydopamine and serum deprivation), whereas micromolar amounts of the fusion proteins were intrinsically toxic to cells. The protective effect was independent from the presence of the mutations A30P and A53T, but no protection occurred when cells were challenged with the proteasome inhibitors lactacystin and MG132. We verified that the protection mechanism required the presence of the C-terminal domain of alpha-syn, as nanomolar amounts of the C-terminal truncated fusion protein TAT alpha-syn (WT[1-97]) failed in preventing H2O2 toxicity. To further characterize the molecular mechanisms at the basis of alpha-syn protection, we investigated the possible involvement of the chaperone protein HSP70 that is widely implicated in neuroprotection. We found that, at nanomolar concentrations, TAT-alpha-syn was able to increase HSP70 protein level, whereas at the micromolar scale, TAT-alpha syn decreased HSP70 at the protein level. These effects on HSP70 were independent from the presence of alpha-syn pathogenetic mutations but required the alpha-syn C-terminal domain. The implications for alpha-syn-mediated neurotoxicity and for PD pathogenesis and progression are discussed. PMID- 15345692 TI - The N-terminal copper-binding domain of the amyloid precursor protein protects against Cu2+ neurotoxicity in vivo. AB - The amyloid precursor protein (APP) contains a Cu binding domain (CuBD) localized between amino acids 135 and 156 (APP135-156), which can reduce Cu2+ to Cu1+ in vitro. The physiological function of this APP domain has not yet being established; nevertheless several studies support the notion that the CuBD of APP is involved in Cu homeostasis. We used APP synthetic peptides to evaluate their protective properties against Cu2+ neurotoxicity in a bilateral intra-hippocampal injection model. We found that human APP135-156 protects against Cu2+-induced neurotoxic effects, such as, impairment of spatial memory, neuronal cell loss, and astrogliosis. APP135-156 lacking two histidine residues showed protection against Cu2+; however, APP135-156 mutated in cysteine 144, a key residue in the reduction of Cu2+ to Cu1+, did not protect against Cu2+ neurotoxicity. In accordance with recent reports, the CuBD of the Caenorhabditis elegans, APL-1, protected against Cu2+ neurotoxicity in vivo. We also found that Cu2+ neurotoxicity is associated with an increase in nitrotyrosine immunofluorescence as well as with a decrease in Cu2+ uptake. The CuBD of APP therefore may play a role in the detoxification of brain Cu. PMID- 15345693 TI - PrPc capping in T cells promotes its association with the lipid raft proteins reggie-1 and reggie-2 and leads to signal transduction. AB - The cellular prion protein (PrPc) resides in lipid rafts, yet the type of raft and the physiological function of PrPc are unclear. We show here that cross linking of PrPc with specific antibodies leads to 1) PrPc capping in Jurkat and human peripheral blood T cells; 2) to cocapping with the intracellular lipid raft proteins reggie-1 and reggie-2; 3) to signal transduction as seen by MAP kinase phosphorylation and an elevation of the intracellular Ca2+ concentration; 4) to the recruitment of Thy-1, TCR/CD3, fyn, lck and LAT into the cap along with local tyrosine phosphorylation and F-actin polymerization, and later, internalization of PrPc together with the reggies into limp-2 positive lysosomes. Thus, PrPc association with reggie rafts triggers distinct transmembrane signal transduction events in T cells that promote the focal concentration of PrPc itself by guiding activated PrPc into preformed reggie caps and then to the recruitment of important interacting signaling molecules. PMID- 15345694 TI - Interleukin-6: a cytokine to forget. AB - It is known that proinflammatory cytokines such as interleukin-6 (IL-6) are expressed in the central nervous system (CNS) during disease conditions and affect several brain functions including memory and learning. In contrast to these effects observed during pathological conditions, here we describe a physiological function of IL-6 in the "healthy" brain in synaptic plasticity and memory consolidation. During long-term potentiation (LTP) in vitro and in freely moving rats, IL-6 gene expression in the hippocampus was substantially increased. This increase was long lasting, specific to potentiation, and was prevented by inhibition of N-methyl-D-aspartate receptors with (+/-)-2-amino-5 phosphonopentanoic acid (AP-5). Blockade of endogenous IL-6 by application of a neutralizing anti-IL-6 antibody 90 min after tetanus caused a remarkable prolongation of LTP. Consistently, blockade of endogenous IL-6, 90 min after hippocampus-dependent spatial alternation learning resulted in a significant improvement of long-term memory. In view of the suggested role of LTP in memory formation, these data implicate IL-6 in the mechanisms controlling the kinetics and amount of information storage. PMID- 15345695 TI - Transplantation of low dose CD34+KDR+ cells promotes vascular and muscular regeneration in ischemic limbs. AB - Hematopoietic progenitor cell transplantation can contribute to revascularization of ischemic tissues. Yet, the optimal cell population to be transplanted has yet to be determined. We have compared the therapeutic potential of two subsets of human cord blood CD34+ progenitors, either expressing the VEGF-A receptor 2 (KDR) or not. In serum-free starvation culture, CD34+KDR+ cells reportedly showed greater resistance to apoptosis and ability to release VEGF-A, as compared with CD34+KDR- cells. When injected into the hind muscles in immunodeficient SCIDbg mice subjected to unilateral ischemia, a low number (10(3)) of CD34+KDR+ cells improved limb salvage and hemodynamic recovery better than a larger dosage (10(4)) of CD34+KDR- cells. The neovascularization induced by KDR+ cells was significantly superior to that promoted by KDR- cells. Similarly, endothelial cell apoptosis and interstitial fibrosis were significantly attenuated by KDR+ cells, which differentiated into mature human endothelial cells and also apparently skeletal muscle cells. This study demonstrates that a low number of CD34+KDR+ cells favors reparative neovascularization and possibly myogenesis in limb ischemia, suggesting the potential use of this cell population in regenerative medicine. PMID- 15345696 TI - Adenosine up-regulation of the mucin gene, MUC2, in asthma. AB - Mucus hypersecretion is a hallmark of asthma that contributes to airway obstruction. While the etiology is not well understood, hypersecretion has been linked to the presence of cytokines such as IL-4, IL-5, IL-9, and IL-13 in the inflamed airway. The presence of adenosine has also been noted in asthmatic airways, and adenosine-mediated signaling in mast cells has been implicated in the severe bronchoconstriction and inflammation prevalent in these patients (1, 2). Here we examine the possibility that adenosine also contributes to mucus hypersecretion by airway epithelial cells. Results in cultured airway epithelial cells showed that MUC2 mucin expression increased in response to adenosine. This appeared to be mediated by a pathway initiated at the adenosine A1 receptor that transduced signals through a Ca2+-activated Cl- channel and EGFR. That this signaling cascade is relevant to asthmatic hypersecretion was indicated by results showing that mucin induction by asthmatic tracheal aspirates was reduced by A1, CLCA1, and EGFR inhibitors. These results suggest that adenosine cooperates with inflammatory cytokines to stimulate mucin production in the asthmatic airway and supports the use of A1, CLCA1, and EGFR inhibitors in the treatment of asthma. PMID- 15345697 TI - Life after corpse engulfment: phagocytosis of apoptotic cells leads to VEGF secretion and cell growth. AB - Removal of apoptotic cells by neighboring viable cells or professional phagocytes is essential for the maintenance of tissue homeostastis. Here we show that the phagocytosis of apoptotic Jurkat T cells by mouse epithelial cells (HC-11) and peritoneal macrophages leads to the secretion of growth and survival factors. We characterized VEGF as one of these factors which subsequently promote the proliferation of endothelial cells. Further we demonstrate that the phagocytosis of apoptotic bodies inhibits both spontanous and UV-irradiation-induced apoptosis in endothelial and epithelial cells. These effects were not observed when phagocytes had been exposed to viable or necrotic Jurkat T cells. We conclude that phagocytosis of apoptotic cells leads to secretion of growth and survival factors by phagocytes that represents a new form of life-promoting cell-cell interaction. PMID- 15345698 TI - Eliminating tobacco use, a continuing challenge. PMID- 15345699 TI - In opposition to Resolution 42. PMID- 15345700 TI - Conclusion of frequently used unsupported by data. PMID- 15345702 TI - Cigarette Smoking Among Adults--United States, 2002. PMID- 15345701 TI - Tobacco dependence curricula in undergraduate osteopathic medical education. AB - CONTEXT: Tobacco use has been identified as the primary preventable cause of premature deaths and disability, yet results of a previous survey show that undergraduate allopathic medical schools do not adequately address this topic. OBJECTIVE: To assess the content and extent of tobacco education and intervention skills in osteopathic medical schools' curricula. DESIGN: A mailed survey with 19 questions similar to one used for allopathic medical schools. SETTING: Nineteen osteopathic medical schools. PARTICIPANTS: Responses were obtained from each associate dean for medical education or representative. MAIN OUTCOME MEASURES: Curriculum in seven basic science and six clinical science content areas (elective or required), hours of tobacco use intervention education, and resource materials used to design curricula. RESULTS: Average number of total content areas covered was 10.2 (6 +/- 1.6 basic science areas, 4.17 +/- 1.54 clinical areas) with a range of 2 to 13. Nine (47%) schools reported covering all seven basic science areas, and one school reported covering none. Eleven (64.7%) of seventeen schools reported less than 3 hours of training in tobacco dependence treatment techniques during all 4 years. Sixty percent of schools do not require clinical training in smoking cessation techniques. Thirty-six percent require clinical training in an artificial setting without patients. None of the schools require clinical training with live patients. The schools founded after 1920 covered an average of almost twice as many content areas as those founded before 1920 (11.1 vs 6.6; P = .018). CONCLUSIONS: Most US osteopathic medical school graduates are not being adequately educated to treat nicotine dependence as recommended by the National Cancer Institute expert panel and the Public Health Service Clinical Practice Guideline. Specifically, osteopathic medical education is deficient in clinical nicotine dependence treatment during the third and fourth years. PMID- 15345703 TI - Cigarette Use Among High School Students--United States, 1991-2003. PMID- 15345704 TI - Relation between variables of preadmission, medical school performance, and COMLEX-USA levels 1 and 2 performance. AB - The purpose of this study was to investigate the relation between preadmission academic variables, osteopathic medical school performance in the first 2 years, and performance on the Comprehensive Osteopathic Medical Licensing Examination (COMLEX-USA) Levels 1 and 2. The study group comprised 174 students in the class of 2001 of the New York College of Osteopathic Medicine of the New York Institute of Technology, Old Westbury. Preadmission academic variables were the Medical College Admission Test (MCAT) subscores and undergraduate grade point averages (UGPAs). Physical sciences (physical MCAT) and biological sciences MCAT (biological MCAT) subscores were significantly correlated with COMLEX-USA Level 1 performance, and verbal reasoning, physical, and biological MCAT subscores were correlated with Level 2 performance. COMLEX-USA Level 1 performance was correlated with the year 1 grade point average (GPA) (0.78) and the year 2 GPA (0.83). COMLEX-USA Level 2 performance was correlated with the year 1 GPA (0.64) and the year 2 GPA (0.68). Strong correlations existed between all year 1 and most year 2 course grades and COMLEX-USA Level 1 scores. School-specific regression models that were highly predictive of school performance and COMLEX USA Level 1 performance were developed. COMLEX-USA Level 1 predictive models used preadmission variables combined with the year 1 and year 2 course grades. The year 2 courses' model had a higher predictive value for COMLEX-USA Level 1 performance (R2 = 0.81) than the year 1 courses' model (R2 = 0.77). Significant predictors of COMLEX-USA Level 1 performance in the combined year 1 and 2 courses' model were the pharmacology II, neuropathology, and pulmonary pathology grades, and the verbal and physical MCAT subscores (R2 = 0.820). PMID- 15345705 TI - Sequence variants of the gene encoding chemoattractant receptor expressed on Th2 cells (CRTH2) are associated with asthma and differentially influence mRNA stability. AB - The gene, CRTH2, encoding a receptor for prostaglandin D(2) (PGD(2)), is located within the peak linkage region for asthma on chromosome (Chr.) 11q reported in African American families. Family-based analysis of asthma and two common SNPs [G1544C and G1651A (rs545659)] in the 3'-untranslated region of CRTH2 showed significant evidence of linkage in the presence of disequilibrium for the 1651G allele (P = 0.003) of SNP rs545659. Haplotype analysis yielded additional evidence of linkage disequilibrium for the 1544G-1651G haplotype (P < 0.001). Population-based case-control analyses were conducted in two independent populations, and demonstrated significant association of the 1544G-1651G haplotype with asthma in an African American population (P = 0.004), and in a population of Chinese children (P < 0.001). Moreover, in the Chinese children the frequency of the 1651G allele in near-fatal asthmatics was significantly higher than mild-to-moderate asthmatics (P = 0.001) and normal controls (P < 0.001). The 1651G allele of SNP re545659 was also associated with a higher degree of bronchial hyperresponsiveness (P < 0.027). Transcriptional pulsing experiments showed that the 1544G-1651G haplotype confers a significantly higher level of reporter mRNA stability, when compared with a non-transmitted haplotype (1544C 1651A), suggesting that the CRTH2 gene on Chr. 11q is a strong candidate gene for asthma. PMID- 15345706 TI - Evidence of novel neuronal functions of dysbindin, a susceptibility gene for schizophrenia. AB - Genetic variation in dysbindin (DTNBP1: dystrobrevin-binding protein 1) has recently been shown to be associated with schizophrenia. The dysbindin gene is located at chromosome 6p22.3, one of the most promising susceptibility loci in schizophrenia linkage studies. We attempted to replicate this association in a Japanese sample of 670 patients with schizophrenia and 588 controls. We found a nominally significant association with schizophrenia for four single nucleotide polymorphisms and stronger evidence for association in a multi-marker haplotype analysis (P = 0.00028). We then explored functions of dysbindin protein in primary cortical neuronal culture. Overexpression of dysbindin induced the expression of two pre-synaptic proteins, SNAP25 and synapsin I, and increased extracellular basal glutamate levels and release of glutamate evoked by high potassium. Conversely, knockdown of endogenous dysbindin protein by small interfering RNA (siRNA) resulted in the reduction of pre-synaptic protein expression and glutamate release, suggesting that dysbindin might influence exocytotic glutamate release via upregulation of the molecules in pre-synaptic machinery. The overexpression of dysbindin increased phosphorylation of Akt protein and protected cortical neurons against neuronal death due to serum deprivation and these effects were blocked by LY294002, a phosphatidylinositol 3 kinase (PI3-kinase) inhibitor. SiRNA-mediated silencing of dysbindin protein diminished Akt phosphorylation and facilitated neuronal death induced by serum deprivation, suggesting that dysbindin promotes neuronal viability through PI3 kinase-Akt signaling. Genetic variants associated with impairments of these functions of dysbindin could play an important role in the pathogenesis of schizophrenia. PMID- 15345707 TI - Mutation in saposin D domain of sphingolipid activator protein gene causes urinary system defects and cerebellar Purkinje cell degeneration with accumulation of hydroxy fatty acid-containing ceramide in mouse. AB - The sphingolipid activator proteins (saposins A, B, C and D) are small homologous glycoproteins that are encoded by a single gene in tandem within a large precursor protein (prosaposin) and are required for in vivo degradation of some sphingolipids with relatively short carbohydrate chains. Human patients with prosaposin or specific saposin B or C deficiency are known, and prosaposin- and saposin A-deficient mouse lines have been generated. Experimental evidence suggests that saposin D may be a lysosomal acid ceramidase activator. However, no specific saposin D deficiency state is known in any mammalian species. We have generated a specific saposin D(-/-) mouse by introducing a mutation (C509S) into the saposin D domain of the mouse prosaposin gene. Saposin D(-/-) mice developed progressive polyuria at around 2 months and ataxia at around 4 months. Pathologically, the kidney of saposin D(-/-) mice showed renal tubular degeneration and eventual hydronephrosis. In the nervous system, progressive and selective loss of the cerebellar Purkinje cells in a striped pattern was conspicuous, and almost all Purkinje cells disappeared by 12 months. Biochemically, ceramides, particularly those containing hydroxy fatty acids accumulated in the kidney and the brain, most prominently in the cerebellum. These results not only indicate the role of saposin D in in vivo ceramide metabolism, but also suggest possible cytotoxicity of ceramide underlying the cerebellar Purkinje cell and renal tubular cell degeneration. PMID- 15345708 TI - Variation in the alcohol content of a 'drink' of wine and spirit poured by a sample of the Scottish population. AB - The standard UK alcohol unit is used to record alcohol consumption and, in health promotion, as a useful yardstick by which the public may be encouraged to monitor their own drinking levels. To investigate the correspondence between this standard unit and the actual amount contained in the 'usual' drink poured by a sample of the Scottish public, participants (n=251) were recruited from three employers in a major city--a manufacturer, an academic and a financial institution. Following a brief questionnaire, participants were asked to pour their usual drink of wine, and then spirit, into a glass. Among drinkers (n=238), the mean amount of alcohol in a drink of wine corresponded to not 1, but 1.92 UK units. For spirit, the corresponding figure was 2.3 UK units. For wine, 43% of the sample poured more than 2 units, for spirit, 55%. (Males poured significantly more spirit than females.) These findings may have important implications for individuals who wish to promote and to adopt sensible drinking practices when consuming wine and spirit at home. Also, the reliability of many consumption surveys, where there is often the implicit assumption that a 'drink' is equivalent to a 'standard unit', must be questioned. PMID- 15345710 TI - The leucine-rich repeat protein LRIG1 is a negative regulator of ErbB family receptor tyrosine kinases. AB - The molecular mechanisms by which mammalian receptor tyrosine kinases are negatively regulated remain largely unexplored. Previous genetic and biochemical studies indicate that Kekkon-1, a transmembrane protein containing leucine-rich repeats and an immunoglobulin-like domain in its extracellular region, acts as a feedback negative regulator of epidermal growth factor (EGF) receptor signaling in Drosophila melanogaster development. Here we tested whether the related human LRIG1 (also called Lig-1) protein can act as a negative regulator of EGF receptor and its relatives, ErbB2, ErbB3, and ErbB4. We observed that in co-transfected 293T cells, LRIG1 forms a complex with each of the ErbB receptors independent of growth factor binding. We further observed that co-expression of LRIG1 with EGF receptor suppresses cellular receptor levels, shortens receptor half-life, and enhances ligand-stimulated receptor ubiquitination. Finally, we observed that co expression of LRIG1 suppresses EGF-stimulated transformation of NIH3T3 fibroblasts and that the inducible expression of LRIG1 in PC3 prostate tumor cells suppresses EGF- and neuregulin-1-stimulated cell cycle progression. Our observations indicate that LRIG1 is a negative regulator of the ErbB family of receptor tyrosine kinases and suggest that LRIG1-mediated receptor ubiquitination and degradation may contribute to the suppression of ErbB receptor function. PMID- 15345711 TI - Reduced beta-amyloid production and increased inflammatory responses in presenilin conditional knock-out mice. AB - Mutations in presenilins (PS) 1 and 2 are the major cause of familial Alzheimer's disease. Conditional double knock-out mice lacking both presenilins in the postnatal forebrain (PS cDKO mice) exhibit memory and synaptic plasticity impairments followed by progressive neurodegeneration in the cerebral cortex. Here we further investigate the molecular events that may underlie the observed phenotypes and identify additional neuropathological markers in the PS cDKO brain. Enzyme-linked immunosorbent assay analysis showed reduced levels of the toxic beta-amyloid (Abeta) peptides in the cerebral cortex of PS cDKO mice. Interestingly, the reduction in Abeta40 and Abeta42 peptides is similar in PS1 conditional knock-out and PS cDKO mice. We further examined the gene expression profile by oligonucleotide microarrays in the PS cDKO cerebral cortex and found that a high number of genes are differentially expressed, most notably a group of up-regulated inflammatory genes. Quantitative real-time reverse transcription PCR and Western analyses confirmed the elevated levels of glial fibrillary acidic protein, complement component C1q, and cathepsin S, up-regulation of which has been associated with inflammatory responses in various neurodegenerative processes. Immunohistochemical analysis revealed that the increase in complement component C1q is confined to the hippocampal formation, whereas glial fibrillary acidic protein and cathepsin S are up-regulated throughout the entire neocortex and hippocampus. In addition, strong microglial activation occurs in the hippocampus and the deeper cortical layers of PS cDKO mice. These results indicate that the memory impairment and neurodegeneration in PS cDKO mice are not caused by Abeta accumulation and that loss of PS function leads to differential up-regulation of inflammatory markers in the cerebral cortex. PMID- 15345712 TI - Crystal structure of severe acute respiratory syndrome coronavirus spike protein fusion core. AB - Severe acute respiratory syndrome coronavirus is a newly emergent virus responsible for a recent outbreak of an atypical pneumonia. The coronavirus spike protein, an enveloped glycoprotein essential for viral entry, belongs to the class I fusion proteins and is characterized by the presence of two heptad repeat (HR) regions, HR1 and HR2. These two regions are understood to form a fusion active conformation similar to those of other typical viral fusion proteins. This hairpin structure likely juxtaposes the viral and cellular membranes, thus facilitating membrane fusion and subsequent viral entry. The fusion core protein of severe acute respiratory syndrome coronavirus spike protein was crystallized, and the structure was determined at 2.8 A of resolution. The fusion core is a six helix bundle with three HR2 helices packed against the hydrophobic grooves on the surface of central coiled coil formed by three parallel HR1 helices in an oblique antiparallel manner. This structure shares significant similarity with the fusion core structure of mouse hepatitis virus spike protein and other viral fusion proteins, suggesting a conserved mechanism of membrane fusion. Drug discovery strategies aimed at inhibiting viral entry by blocking hairpin formation, which have been successfully used in human immunodeficiency virus 1 inhibitor development, may be applicable to the inhibition of severe acute respiratory syndrome coronavirus on the basis of structural information provided here. The relatively deep grooves on the surface of the central coiled coil will be a good target site for the design of viral fusion inhibitors. PMID- 15345713 TI - Nuclear import of proinflammatory transcription factors is required for massive liver apoptosis induced by bacterial lipopolysaccharide. AB - Stimulation of macrophages with lipopolysaccharide (LPS) leads to the production of cytokines that elicit massive liver apoptosis. We investigated the in vivo role of stress-responsive transcription factors (SRTFs) in this process focusing on the precipitating events that are sensitive to a cell-permeant peptide inhibitor of SRTF nuclear import (cSN50). In the absence of cSN50, mice challenged with LPS displayed very early bursts of inflammatory cytokines/chemokines, tumor necrosis factor alpha (1 h), interleukin 6 (2 h), interleukin 1 beta (2 h), and monocyte chemoattractant protein 1 (2 h). Activation of both initiator caspases 8 and 9 and effector caspase 3 was noted 4 h later when full-blown DNA fragmentation and chromatin condensation were first observed (6 h). At this time an increase of pro-apoptotic Bax gene expression was observed. It was preceded by a decrease of anti-apoptotic Bcl2 and BclX(L) gene transcripts. Massive apoptosis was accompanied by microvascular injury manifested by hemorrhagic necrosis and a precipitous drop in blood platelets observed at 6 h. An increase in fibrinogen/fibrin degradation products and a rise in plasminogen activator inhibitor 1 occurred between 4 and 6 h. Inhibition of SRTFs nuclear import with the cSN50 peptide abrogated all these changes and increased survival from 7 to 71%. Thus, the nuclear import of SRTFs induced by LPS is a prerequisite for activation of the genetic program that governs cytokines/chemokines production, liver apoptosis, microvascular injury, and death. These results should facilitate the rational design of drugs that protect the liver from inflammation-driven apoptosis. PMID- 15345714 TI - Polyglutamine expansion in ataxin-3 does not affect protein stability: implications for misfolding and disease. AB - Polyglutamine proteins that cause neurodegenerative disease are known to form proteinaceous aggregates, such as nuclear inclusions, in the neurons of affected patients. Although polyglutamine proteins have been shown to form fibrillar aggregates in a variety of contexts, the mechanisms underlying the aberrant conformational changes and aggregation are still not well understood. In this study, we have investigated the hypothesis that polyglutamine expansion in the protein ataxin-3 destabilizes the native protein, leading to the accumulation of a partially unfolded, aggregation-prone intermediate. To examine the relationship between polyglutamine length and native state stability, we produced and analyzed three ataxin-3 variants containing 15, 28, and 50 residues in their respective glutamine tracts. At pH 7.4 and 37 degrees C, Atax3(Q50), which lies within the pathological range, formed fibrils significantly faster than the other proteins. Somewhat surprisingly, we observed no difference in the acid-induced equilibrium and kinetic un/folding transitions of all three proteins, which indicates that the stability of the native conformation was not affected by polyglutamine tract extension. This has led us to reconsider the mechanisms and factors involved in ataxin-3 misfolding, and we have developed a new model for the aggregation process in which the pathways of un/folding and misfolding are distinct and separate. Furthermore, given that native state stability is unaffected by polyglutamine length, we consider the possible role and influence of other factors in the fibrillization of ataxin-3. PMID- 15345715 TI - The tumor suppressor p53 abrogates Smad-dependent collagen gene induction in mesenchymal cells. AB - The pleiotropic cytokine transforming growth factor-beta (TGF-beta) is a potent inducer of collagen synthesis and is implicated in the pathogenesis of fibrosis. Acting in concert with transcriptional coactivators p300/CBP, the Smads mediate TGF-beta stimulation of collagen synthesis in human dermal fibroblasts. Little information exists regarding positive and negative modulation of physiological TGF-beta responses. Because the tumor suppressor p53 is implicated in connective tissue homeostasis, here we examined the regulation of collagen gene expression by p53. Forced expression of ectopic p53 in dermal fibroblasts repressed basal and TGF-beta-stimulated collagen gene expression, whereas the absence of cellular p53 was associated with significantly enhanced transcriptional activity of the Type I collagen gene (COL1A2) and collagen synthesis. Ectopic expression of p53 also repressed TGF-beta stimulation of promoter activity driven by minimal Smad binding elements, suggesting that p53 modulated Smad-dependent intracellular signaling. Inhibition was not due to altered levels, phosphorylation, or nuclear translocation of cellular Smads. Treatment of fibroblasts with etoposide, a potent inducer of cellular p53, abrogated TGF-beta stimulation of COL1A2 promoter activity and collagen synthesis in a p53-dependent manner. Overexpression of the transcriptional coactivator p300 rescued TGF-beta stimulation of COL1A2 promoter activity in fibroblasts overexpressing p53. Furthermore, the ligand-induced interaction of cellular Smad3 with p300 or with its cognate Smad-binding DNA element and recruitment of p300 to the DNA-protein complex assembled on the Smad binding element were markedly reduced in p53-overexpressing fibroblasts. Collectively, these results indicate, for the first time, that p53 is a potent and selective endogenous repressor of TGF-beta-regulated collagen gene expression in dermal fibroblasts. The ligand-dependent interaction of Smad3 with p300 may be one of the targets of p53-mediated inhibition of TGF-beta responses. These findings suggest that a novel and important physiologic function for the tumor suppressor p53 is the regulation of fibrotic cellular responses. PMID- 15345716 TI - Rapid plasma membrane-endosomal trafficking of the lymph node sinus and high endothelial venule scavenger receptor/homing receptor stabilin-1 (FEEL-1/CLEVER 1). AB - The sinusoidal endothelia of liver, spleen, and lymph node are major sites for uptake and recycling of waste macromolecules through promiscuous binding to a disparate family of scavenger receptors. Among the most complex is stabilin-1, a large multidomain protein containing tandem fasciclin domains, epidermal growth factor-like repeats, and a C-type lectin-like hyaluronan-binding Link module, which functions as an endocytic receptor for acetylated low density lipoprotein and advanced glycation end products. Intriguingly, stabilin-1 has also been reported to mediate both homing of leukocytes across lymph node high endothelial venules and adhesion of metastatic tumor cells to peritumoral lymphatic vessels. Currently, however, it is not clear how stabilin-1 mediates these distinct functions. To address the issue, we have investigated the tissue and subcellular localization of stabilin-1 in detail and assessed the functional status of its Link module. We show that stabilin-1 is almost entirely intracellular in lymph node high endothelial venules, lymphatic sinus endothelium, and cultured endothelial cells but that a finite population, detectable only by fluorescent antibody or fluorescein-labeled (Fl)-acetylated low density lipoprotein uptake, cycles rapidly between the plasma membrane and EEA-1+ve (early endosome antigen 1) early endosomes. In addition, we show using full-length stabilin-1 cDNA and a stabilin-1/CD44 chimera in HeLa cells that intracellular targeting is influenced by the transmembrane domain/cytoplasmic tail, which contains a putative dileucine (DXXLL) Golgi to endosomal sorting signal. Finally, we provide evidence that the stabilin-1 Link domain binds neither hyaluronan nor other glycosaminoglycans. These properties support a role for stabilin-1 as a rapidly recycling scavenger receptor and argue against a role in cell adhesion or lymphocyte homing. PMID- 15345717 TI - Integrin activation state determines selectivity for novel recognition sites in fibrillar collagens. AB - Only three recognition motifs, GFOGER, GLOGER, and GASGER, all present in type I collagen, have been identified to date for collagen-binding integrins, such as alpha(2)beta(1). Sequence alignment was used to investigate the occurrence of related motifs in other human fibrillar collagens, and located a conserved array of novel GER motifs within their triple helical domains. We compared the integrin binding properties of synthetic triple helical peptides containing examples of such sequences (GLSGER, GMOGER, GAOGER, and GQRGER) or the previously identified motifs. Recombinant inserted (I) domains of integrin subunits alpha(1), alpha(2) and alpha(11) all bound poorly to all motifs other than GFOGER and GLOGER. Similarly, alpha(2)beta(1) -containing resting platelets adhered well only to GFOGER and GLOGER, while ADP-activated platelets, HT1080 cells and two active alpha(2)I domain mutants (E318W, locked open) bound all motifs well, indicating that affinity modulation determines the sequence selectivity of integrins. GxO/SGER peptides inhibited platelet adhesion to collagen monomers with order of potency F >/= L >/= M > A. These results establish GFOGER as a high affinity sequence, which can interact with the alpha(2)I domain in the absence of activation and suggest that integrin reactivity of collagens may be predicted from their GER content. PMID- 15345718 TI - Identification of SNF1/AMP kinase-related kinase as an NF-kappaB-regulated anti apoptotic kinase involved in CD95-induced motility and invasiveness. AB - The death receptor CD95 (APO-1/Fas) induces apoptosis in many tissues. However, in apoptosis-resistant tumor cells, stimulation of CD95 induces up-regulation of a defined number of mostly anti-apoptotic genes, resulting in increased motility and invasiveness of tumor cells. The majority of these genes are known NF-kappaB target genes. We have identified one of the CD95-regulated genes as the serine/threonine kinase (SNF1/AMP kinase-related kinase (SNARK)), which is induced in response to various forms of metabolic stress. We demonstrate that up regulation of SNARK in response to CD95 ligand and tumor necrosis factor alpha depends on activation of NF-kappaB. Overexpression of SNARK rendered tumor cells more resistant, whereas a kinase-inactive mutant of SNARK sensitized cells to CD95-mediated apoptosis. Furthermore, small interfering RNA-mediated knockdown of SNARK increased the sensitivity of tumor cells to CD95 ligand- and TRAIL-induced apoptosis. Importantly, cells with reduced expression of SNARK also showed reduced motility and invasiveness in response to CD95 engagement. SNARK therefore represents an NF-kappaB-regulated anti-apoptotic gene that contributes to the tumor-promoting activity of CD95 in apoptosis-resistant tumor cells. PMID- 15345719 TI - Gbetagamma activation of Src induces caveolae-mediated endocytosis in endothelial cells. AB - Caveolae-mediated endocytosis in endothelial cells is stimulated by the binding of albumin to gp60, a specific albumin-binding protein localized in caveolae. The activation of gp60 induces its cell surface clustering and association with caveolin-1, the caveolar-scaffolding protein. This interaction leads to G(i) induced Src kinase activation, which in turn signals dynamin-2-mediated fission and directed migration of caveolae-derived vesicles from apical to basal membrane. In this study, we investigated the possible role of the Gbetagamma heterodimer in signaling G(i)-induced Src activation and subsequent caveolae mediated endocytosis. We observed using rat lung microvascular endothelial cells that expression of the C terminus of beta-adrenergic receptor kinase (ct betaARK), an inhibitor Gbetagamma signaling, prevented gp60-dependent Src activation as well as caveolae-mediated endocytosis and transcellular transport of albumin and uptake of cholera toxin subunit B, a specific marker of caveolae internalization. Expression of ct-betaARK also prevented Src-mediated tyrosine phosphorylation of caveolin-1 and dynamin-2 and the resultant phosphorylation dependent association of dynamin-2 and caveolin-1. Also, the direct activation of Gbetagamma using a specific cell-permeant activating peptide (myristoylated SIRKALNILGYPDYD) simulated the effects of gp60 in inducing Src activation, caveolin-1, and dynamin-2 phosphorylation as well as caveolae-mediated endocytosis of cholera toxin subunit B. The myristoylated-SIRKALNILGYPDYD peptide induced responses were inhibited by the expression of ct-betaARK. Taken together, our results demonstrate that Gbetagamma activation of Src signals caveolae mediated endocytosis and transendothelial albumin transport via transcytosis. PMID- 15345720 TI - Identification of a novel epitope in the thyroid-stimulating hormone receptor ectodomain acting as intramolecular signaling interface. AB - Glycoprotein hormone receptors (GPHRs) differ from the other seven transmembrane receptors mainly through a complex activation mechanism that requires the binding of a large hormone toward a large N-terminal ectodomain. The intramolecular mechanism of the signal transduction to the serpentine domain upon hormone binding at the ectodomain is not understood. To identify determinants at the GPHR ectodomain that may be involved in signal transduction, we first searched for homologous structural features. Based on high sequence similarity to the determined structures of the Nogo-receptor ectodomain and the intermolecular complex of the Interleukin-8 ligand (IL8) and the N-terminal peptide of the IL8 receptor (IL8RA), the hypothesis was developed that portions of the intramolecular components, Cysteine-box-2 and Cysteine-box-3, of the GPHR ectodomain interact and localize at the interface between ectodomain and serpentine domain. Indeed, point mutations within the D403EFN406 motif at Cysteine-box-3 of the thyrotropin receptor resulted in increased basal cAMP levels, suggesting that this motif may be important for transduction of the signal from the ectodomain to the transmembrane domain. New indications are provided about the tight spatial cooperation and relative location of the new epitope and other determinants at the thyrotropin receptor ectodomain, such as the leucine-rich repeat motif Ser281 and the cysteine boxes. According to the high sequence conservation, the results are of general relevance for the signal transduction mechanism of other glycoprotein hormone receptors such as choriogonadotrophic/luteinizing hormone receptor and follicle-stimulating hormone receptor. PMID- 15345721 TI - The inhibitor-1 C terminus facilitates hormonal regulation of cellular protein phosphatase-1: functional implications for inhibitor-1 isoforms. AB - Inhibitor-1 (I-1) is a selective inhibitor of protein phosphatase-1 (PP1) and regulates several PP1-dependent signaling pathways, including cardiac contractility and regulation of learning and memory. The human I-1 gene has been spliced to generate two alternative mRNAs, termed I-1alpha and I-1beta, encoding polypeptides that differ from I-1 in their C-terminal sequences. Reverse transcription-PCR established that I-1alpha and I-1beta mRNAs are expressed in a developmental and tissue-specific manner. Functional analysis of I-1 in a Saccharomyces cerevisiae strain dependent on human I-1 for viability established that a novel domain encompassing amino acids 77-110 is necessary for PP1 inhibition in yeast. Expression of human I-1 in S. cerevisiae with a partial loss of-function eukaryotic initiation factor-2alpha (eIF2alpha) kinase (Gcn2p) mutation permitted growth during amino acid starvation, consistent with the inhibition of Glc7p/PP1, the yeast eIF2alpha phosphatase. In contrast, human I 1alpha, which lacks amino acids 83-134, and I-1 with C-terminal deletions were significantly less effective in promoting yeast growth under starvation conditions. These data suggest that C-terminal sequences of I-1 enhance regulation of the eukaryotic eIF2alpha phosphatase. In vitro studies established that C-terminal sequences, deleted in both I-1alpha and I-1beta, enhance PP1 binding and inhibition. Expression of full-length and C-terminally truncated I-1 in HEK293T cells established the importance of the I-1 C terminus in transducing cAMP signals that promote eIF2alpha phosphorylation. This study demonstrates that multiple domains in I-1 target cellular PP1 complexes and establishes I-1 as a cellular regulator of eIF2alpha phosphorylation. PMID- 15345722 TI - In vivo evidences that insulin regulates human polymorphonuclear neutrophil functions. AB - Polymorphonuclear neutrophils (PMN) are able to destroy invasive mircoorganisms by a wide variety of functions. Whereas insulin does not stimulate hexose transport in PMN, previous reports have clearly shown that this hormone regulates glucose metabolism inside this cell, raising the question of insulin action on PMN functions in humans. It is interesting that in vitro studies established a strong relationship between specific binding of insulin to its PMN membrane receptor and the activation of the main PMN functions. Therefore, investigation in healthy subjects under strict euglycemia and physiological insulinemia was performed to understand the in vivo-specific action of insulin on PMN functions without hyperglycemia interferences. We determined numerous PMN functions before and after hyperinsulinemia (0.5 mU/kg/min) and euglycemia (0.9 g/l) clamp for 4 h in eight adult healthy volunteers (24+/-6 years). The total number of PMN and the number of PMN expressing CD11b, CD15, CD62L, and CD89 were significantly increased over baseline (P<0.001), whereas the density of these receptors was down-regulated (P<0.01) by insulin. PMN chemotaxis (+117%, P<0.05), phagocytosis (+29%, P<0.001), and bactericidal (+17-25%, P<0.001) capacities were increased during the insulin clamp (P<0.05). Therefore, insulin treatment may modulate PMN functions not only by attainment of a better metabolic control, as suggested by in vivo studies in diabetic patients, but also through a direct effect of insulin. PMID- 15345723 TI - G-CSF-mobilized CD34+ cells cultured in interleukin-2 and stem cell factor generate a phenotypically novel monocyte. AB - To study the early stages of development from stem cells of the CD56+ cell population [which includes natural killer (NK) cells], granulocyte-colony stimulating factor-mobilized peripheral blood CD34+ cells from healthy donors were sorted to >99% purity and cultured in the presence of stem cell factor and interleukin (IL)-2. After 3 weeks in culture, the majority of cells acquired CD33, with or without human leukocyte antigen-DR and CD14. In 20 stem cell donors tested, 8.7 +/- 8.8% of cells were CD56+. Two major CD56+ subsets were identified: CD56(bright), mainly CD33- cells (7+/-10%, n=11) with large, granular lymphocyte morphology, and CD56dim, mainly CD33+ (2.5+/-2, n=11) cells with macrophage morphology. The CD56bright population had cytoplasmic granzyme A but lacked killer inhibitory receptor, suggesting they were immature NK cells. The CD56dim, CD33+, population lacked NK markers. They may represent a minor subset of normal monocytes at a developmental stage comparable with the rare CD56+ CD33+ hybrid myeloid/NK cell leukemia. Consistent with a monocyte nature, CD56dimCD33+ proliferated and produced a variety of cytokines upon lipopolysaccharide stimulation, including IL-8, IL-6, monocyte chemoattractant protein-1, and macrophage-derived chemokine but not interferon-gamma. In a short-term cytotoxicity assay, they failed to kill but powerfully inhibited the proliferation of the NK-resistant cell line P815. The generation of CD56+ cells was negatively regulated by hyaluronic acid and IL-4, indicating that extracellular matrix may play an important role in the commitment of CD34+ cells into CD56 myeloid and lymphoid lineages. PMID- 15345725 TI - Emerging technologies: trendy RNA tools for aging research. AB - Aging is an inevitable biological phenomenon. Attempts to understand its mechanisms and, consequently, to therapeutically decelerate or even reverse the process are limited by its daunting complexity. Rapid and robust functional genomic tools suited to a wide array of experimental model systems are needed to dissect the interplay of individual genes during aging. In this article, we review principles that transcend the view of RNA, from a molecule merely mediating the flow of genetic information, into a unique molecular tool. In the form of catalytic molecular scissors (ribozymes), antibody-like antagonists (aptamers) and gene silencers (interfering RNAs, RNAi) can be effectively used to dissect biofunctions conserved throughout the evolution. In this review, application of recent RNA tools in aging research is discussed. PMID- 15345724 TI - Stabilin-1 localizes to endosomes and the trans-Golgi network in human macrophages and interacts with GGA adaptors. AB - Stabilin-1 and stabilin-2 constitute a novel family of fasciclin domain containing hyaluronan receptor homologues recently described by us. Whereas stabilin-1 is expressed in sinusoidal endothelial cells and in macrophages in vivo, stabilin-2 is absent from the latter. In the present study, we analyzed the subcellular distribution of stabilin-1 in primary human macrophages. Using flow cytometry, expression of stabilin-1 was demonstrated on the surface of interleukin-4/dexamethasone-stimulated macrophages (MPhi2). By immunofluorescence and confocal microscopy, we established that stabilin-1 is preferentially localized in early endosome antigen-1-positive early/sorting endosomes and in recycling endosomes identified by transferrin endocytosis. Association of stabilin-1 was infrequently seen with p62 lck ligand-positive late endosomes and with CD63-positive lysosomes but not in lysosome-associated membrane protein-1 positive lysosomes. Stabilin-1 was also found in the trans-Golgi network (TGN) but not in Golgi stack structures. Glutathione S-transferase pull-down assay revealed that the cytoplasmic tail of stabilin-1 but not stabilin-2 binds to recently discovered Golgi-localized, gamma-ear-containing, adenosine 5' diphosphate-ribosylation factor-binding (GGA) adaptors GGA1, GGA2, and GGA3 long, mediating traffic between Golgi and endosomal/lysosomal compartments. Stabilin-1 did not bind to GGA3 short, which lacks a part of the Vps27p/Hrs/STAM domain. Deletion of DDSLL and LL amino acid motifs resulted in decreased binding of stabilin-1 with GGAs. A small portion of stabilin-1 colocalized with GGA2 and GGA3 in the TGN in MPhi2. Treatment with brefeldin A resulted in accumulation of stabilin-1 in the TGN. Our results suggest that stabilin-1 is involved in the GGA mediated sorting processes at the interface of the biosynthetic and endosomal pathways; similarly to other GGA-interacting proteins, stabilin-1 may thus function in endocytic and secretory processes of human macrophages. PMID- 15345726 TI - Divergent effects of caloric restriction on gene expression in normal and long lived mice. AB - Long-lived Ames dwarf mice share many phenotypic characteristics with animals subjected to caloric restriction (CR) but they are not CR mimetics. CR prolongs longevity in both normal and Ames dwarf mice. Using real-time polymerase chain reaction and western blot, we have examined the expression of genes related to insulin signaling in the liver of normal and dwarf mice subjected to 30% CR. The results revealed divergent responses of dwarf and normal animals to CR raising an interesting possibility that CR may affect longevity of normal and dwarf mice by different mechanisms. Moreover, effects of dwarfism on the expression of the examined genes differed from the effects of CR, thus adding to the evidence that these long-lived mutants are not CR mimetics. PMID- 15345727 TI - How much should we eat? The association between energy intake and mortality in a 36-year follow-up study of Japanese-American men. AB - Energy restriction extends life span and lowers mortality from age-related diseases in many species, but the effects in humans are unknown. We prospectively examined this relationship in a large epidemiological study of Japanese-American men. We followed 1915 healthy nonsmokers, aged 45-68 years at study onset, for 36 years. Twenty-four-hour recall of diet was recorded at baseline, and follow-up was for all-cause mortality. After adjustment for age and other confounders, there was a trend toward lower mortality in the second quintile of energy intake, suggesting that men who consumed 15% below the group mean were at the lowest risk for all-cause mortality. Increased mortality was seen with intakes below 50% of group mean. Thus, we observed trends between low energy intake and reduced risk for all-cause mortality in humans until energy intake fell to less than half the group mean, consistent with previous findings in other species. PMID- 15345728 TI - Dual-energy x-ray absorptiometry-measured lean soft tissue mass: differing relation to body cell mass across the adult life span. AB - Lean soft tissue (LST) measured by dual-energy X-ray absorptiometry (DEXA) is used as a metabolic measure in aging research despite evidence of extracellular fluid expansion and a corresponding reduction in body cell mass (BCM) in older participants. We investigated the hypothesis that the fraction of LST as BCM is smaller with greater age. Men and women (n = 2043) had DEXA and 40K-counting for body potassium and BCM measured on the same day. Both BCM and LST were lower with greater age but the relative lowering was larger for BCM. A multiple linear regression model was fitted with BCM/LST as the dependent variable, and age, sex, and interaction terms as independent variables. Men had a mean BCM/LST greater (p < .001) than women; quadratic and cubic age terms were also significant or approached significance. Thus, the fraction of LST as BCM is smaller in older adults, a finding that has implications for the interpretation of DEXA results. PMID- 15345729 TI - Oxygen consumption during maximal exercise in Fischer 344 x Brown Norway F1 hybrid rats. AB - We characterized O2 consumption (VO2) during treadmill exercise in 12-, 24-, and 35-month-old Fischer 344 x Brown Norway F1 hybrid (F344BNF1) rats. When accounting for differences in body mass (Mb), (O2)peak decreased by 10% and 33% in 24- and 35-month-old rats, respectively, compared with rats at 12 months (analysis of covariance, p < .01). O2 cost per unit work at VO2peak (i.e., VO2peak/work) was greater in 35-month-old rats compared with 12- and 24-month-old rats (p < .001). During submaximal exercise, the O2 cost was greater in 24- and 35-month-old than 12-month-old rats (p < .01). Analysis of covariance revealed similar patterns irrespective of differences in Mb or lean Mb as covariates. The underlying mechanism responsible for increasing O2 consumption in aged F344BNF1 rats during exercise, although partly explained by mechanical inefficiencies of locomotion, still remains to be determined. PMID- 15345730 TI - Dehydration and delirium--not a simple relationship. PMID- 15345731 TI - Bioelectrical impedance analysis and other hydration parameters as risk factors for delirium in rural nursing home residents. AB - BACKGROUND: The study investigators conducted a vigorous screening protocol for delirium in rural long-term care (LTC) facilities for a period of 28 days focusing on Bioelectrical Impedance Analysis (BIA) and other hydration parameters as risk factors. METHODS: A two-stage cluster sampling procedure was used to randomly select participants (n = 313) from 13 LTC facilities located in southeastern Iowa, stratified on facility bed size. BIA was used to estimate intracellular water (ICW), extracellular water (ECW), and total body water (TBW) on four occasions--baseline and follow-up days 7, 14, and 28. Volume estimates were calculated as a percent of body weight (%WT). Serum electrolytes and hematology were also measured. Delirium was measured with four strict criteria: a NEECHAM Confusion Scale score < 25, Vigilance "A" score > 2, a Mini-Mental Status Examination < baseline, and a positive Confusion Assessment Method score. RESULTS: There were n = 69 delirium cases (22.0%). Blood urea nitrogen/creatinine ratios greater than 21:1 (odds ratio = 1.76, 95% confidence interval 1.02-3.06). No significant risk for delirium was associated with ICW, ECW, or TBW as a percent of body weight. CONCLUSIONS: Some changes were observed with a slight decrease in ICW between day 7 and day 14 of follow-up that tended to follow an increase in delirium events, but in general the BIA measures did not predict delirium events. PMID- 15345732 TI - Cerebral white matter changes and geriatric syndromes: is there a link? AB - Cerebral white matter lesions (WMLs), also called "leukoaraiosis," are common neuroradiological findings in elderly people. WMLs are often located at periventricular and subcortical areas and manifest as hyperintensities in magnetic resonance imaging. Recent studies suggest that cardiovascular risk factors are associated with the development of WMLs. These lesions are associated with different geriatric syndromes such as falls, executive cognitive impairment, depressive symptoms, and urinary incontinence. Damage to associative pathways in frontal and subcortical regions due to hypoperfusion may disrupt frontal executive, motor control, and other systems, resulting in these manifestations. WMLs are associated with substantial disability and should not be considered a benign and silent condition as once believed. Interventions addressing cardiovascular risk factors should be undertaken in early or mid-life in order to prevent late-life functional impairment associated with WMLs. After these lesions develop and impair executive cognitive functions, the patient's ability to comply with a complex risk reduction program may be significantly compromised. PMID- 15345733 TI - The effect of different doses of micronized 17beta-estradiol on C-reactive protein, interleukin-6, and lipids in older women. AB - BACKGROUND: The authors evaluated the effect of 3 doses (0.25 mg/day, 0.5 mg/day, and 1 mg/day) of micronized 17beta-estradiol (E2) on C-reactive protein (CRP), interleukin-6 (IL-6), and lipids, compared with placebo, in healthy older women participating in an osteoporosis study. METHODS: This randomized, double-blind, placebo-controlled study was conducted in a University clinical research center. Participants were healthy, community-living women older than 65 years. The primary outcome measure of the study was bone metabolism as estimated by serum and urine markers of bone turnover. For this analysis, the authors measured serum markers of CRP, IL-6, lipids, intracellular adhesion molecule-1, and E-selectin at baseline, after 12 weeks of treatment, and after 12 weeks with no treatment. RESULTS: A significant dose-response effect of estrogen occurred on CRP levels. After 12 weeks of treatment, CRP decreased 59% in the 0.25 mg/day E2 group and increased 65% in the 1 mg/day E2 group, compared with placebo. The CRP level continued to be elevated (92%), compared with placebo, 12 weeks after treatment was discontinued in the 1 mg/day E2 group. High-density lipoprotein (HDL) and HDL2 cholesterol increased and low-density lipoprotein (LDL) cholesterol decreased at 12 weeks in the 1 mg/day E2 group, with a significant dose-response effect. E-selectin decreased significantly in the 1 mg/day E2 group 12 weeks after discontinuation of treatment (-7%), and there was a significant dose response effect at this time. The 2 lower doses did not affect any of these parameters. Total and HDL3 cholesterol, triglycerides, lipoprotein(a), intracellular adhesion molecule-1, and IL-6 did not change with any dose of E2. CONCLUSIONS: C-reactive protein, an inflammation marker associated with increased risk for cardiovascular disease, decreased in women taking the lowest estrogen dose but increased in women assigned to the highest estrogen dose, suggesting decreased inflammation with lower dose E2. However, with 3 months of treatment, 0.25 or 0.5 mg/day E2 did not have the same beneficial effects on HDL or LDL cholesterol as did 1 mg/day E2. These data suggest that estradiol doses have differential short-term effects on markers of cardiovascular disease. Low-dose E2 decreased CRP, an important marker of inflammation, but did not affect lipid parameters, whereas the highest dose increased CRP and had a beneficial effect on lipid parameters. The long-term consequences of these effects are unknown, but it is possible that estradiol dose should be considered when risk:benefit ratios are evaluated for individual women before estrogen replacement therapy is initiated. PMID- 15345734 TI - Physical activity and mortality in frail, community-living elderly patients. AB - BACKGROUND: The authors describe the prevalence of moderate-intensity physical activity in a population of older persons living in the community. In addition, they explore the relationship between physical activity and mortality. METHODS: In this longitudinal observational study, the authors analyzed data from patients admitted to home care programs collected as part of the Italian Silver Network Home Care project. Twelve home health agencies participated in the project, which evaluated the implementation of the Minimum Data Set for Home Care (MDS-HC) instrument. A total of 2757 patients were enrolled in the current study. The primary outcome measures were the prevalence of 2 or more hours per week of physical activity and survival. RESULTS: Fewer than 20% of patients had regular physical activity. During a median follow-up period of 10 months from the initial MDS-HC assessment, 442 (16%) patients died. After adjusting for sex, physical and cognitive disability, and all potential risk factors for death, active patients were less likely to die compared with those with no or very low-intensity physical activity (relative risk ratio [RR], 0.51; 95% confidence interval [CI], 0.35-0.73). This inverse relationship was also significant in patients aged 80 years and older (RR 0.55; 95% CI, 0.32-0.95). CONCLUSIONS: Physical activity is associated with a significantly lower risk of all-cause mortality. The current findings support the possibility that moderate-intensity physical activity has an independent effect on survival even among frail and old persons. PMID- 15345735 TI - Activities of daily living function and disability in older adults in a randomized trial of the health enhancement program. AB - BACKGROUND: Disability in basic activities of daily living (ADLs) implies a loss of independence and increases the risk for hospitalization, nursing home admission, and death. Little is known about ways by which ADL disability can be prevented or reversed. The authors evaluated the efficacy of the Health Enhancement Program in preventing and reducing ADL disability in community dwelling older adults. METHODS: The authors analyzed data from a 12-month, randomized, single-blinded, controlled trial of a disability prevention, chronic disease self-management program involving 201 adults aged 70 years and older that was conducted from February 1995 to June 1996 at a senior center in western Washington state. Activities of daily living disability incidence, improvement, and worsening were assessed using intention-to-treat methods. RESULTS: The cumulative incidence of ADL disability among those who were not ADL disabled at baseline (n = 56 in the intervention group, n = 57 in the control group) was modestly lower in the intervention group than in the control group at 12 months (14.3% vs 21.3%, p = .466). Cumulative improvement in ADL function among those who reported any ADL disability at baseline (n = 41 in the intervention group, n = 43 in the control group) was greater in the intervention group at 12 months (80.5% vs 46.5%, p = .026). The likelihood for ADL improvement was greater in the intervention group compared with controls at 12 months (adjusted hazard ratio, 1.84; 95% confidence interval, 1.05 to 3.22; p = .020). Cumulative worsening of ADL function was slightly lower in the intervention group at 12 months (18.6% vs 26.5%, p = .237). Intervention participants tended to be at lower risk for ADL worsening (adjusted hazard ratio, 0.71; 95% confidence interval, 0.38 to 1.30; p = .266) compared with control participants. CONCLUSION: The Health Enhancement Program intervention led to improved ADL functioning in those who were disabled initially and thereby offers a promising strategy for limiting or reversing functional decline in disabled elderly persons. PMID- 15345736 TI - Naps and circadian rhythms in postmenopausal women. AB - BACKGROUND: Napping patterns and relationships among naps, circadian rhythms, and nocturnal sleep were evaluated in postmenopausal women. Naps and nocturnal sleep were compared between depressed women and healthy volunteers. METHODS: The study included 436 postmenopausal women between the ages of 50 and 81 years. Psychiatric interviews were conducted using the Structured Clinical Interview for DSM-IV Axis I Disorders. Each participant wore a wrist-activity monitor and kept a daily sleep log at home for 1 week. The major urinary melatonin metabolite (6 sulfatoxymelatonin [aMT6s]) was measured for two 24-hour periods at home. RESULTS: There was no significant difference in naps and nocturnal sleep patterns between depressed participants (n = 30) and those with no history of mental disorder (n = 222). Three peaks occurred in the timing of napping: approximately 1 hour before bedtime, 8 to 9 hours after wake-up time, and 2 hours after waking, in descending order of magnitude. Significant inverse correlations were observed between evening nap duration and both wake-up time and aMT6s acrophase. The onset time of aMT6s excretion was advanced by 32 minutes in evening nappers. CONCLUSIONS: Evening napping, a characteristic of these postmenopausal women, may be related to an advance of circadian rhythms in this age group. Relationships between evening napping and earlier wake-up time may be a common manifestation of advanced circadian rhythms or may be a homeostatic effect. PMID- 15345737 TI - The effects of serum beta-carotene concentration and burden of inflammation on all-cause mortality risk in high-functioning older persons: MacArthur studies of successful aging. AB - BACKGROUND: It remains unclear to what extent the associations between low serum beta-carotene concentration and increased risk for cardiovascular disease and cancers are attributable to inflammation. The objective of this study was to evaluate simultaneously the effects of serum beta-carotene concentration and inflammation on the subsequent all-cause mortality risk in high-functioning older persons. METHODS: The authors conducted a prospective cohort study using information from 672 participants from the MacArthur Studies of Successful Aging. Baseline information was obtained for serum concentrations of beta-carotene, C reactive protein, interleukin-6, cholesterols, and albumin; body mass index; waist:hip ratio; prevalent medical conditions; health behaviors; and medications. Sex-specific univariate and multivariate logistic regression analyses were used to study the effects of low beta-carotene, high inflammation burden, or both on 7 year all-cause mortality rates while adjusting for other confounders. RESULTS: The serum beta-carotene concentration was inversely associated with C-reactive protein and interleukin-6 levels. After adjustment for inflammation markers and other covariates, the relative risks for low beta-carotene for the 7-year all cause mortality risk were 2.30 (95% confidence interval [CI], 1.23 to 4.31) in men and 0.85 (95% CI, 0.42 to 1.75) in women. Compared with men with high beta carotene levels and low inflammation, the multiply adjusted relative risk for low beta-carotene and high inflammation burden was 3.78 (95% CI, 1.69 to 8.47) in men. CONCLUSIONS: Low levels of serum beta-carotene are independently associated with an increased all-cause mortality risk in older men, even after adjustment for the effects of inflammation and other risk factors. In men, but not women, a synergistic effect occurs between low beta-carotene concentration and high inflammation burden in predicting higher mortality rates. PMID- 15345738 TI - Effects of megestrol acetate on circulating interleukin-15 and interleukin-18 concentrations in healthy elderly men. AB - BACKGROUND: Interleukin-15 (IL-15) and interleukin-18 (IL-18) are potential regulators of body composition in humans. The authors previously reported that megestrol acetate ingestion causes a large accumulation of adipose tissue and reduces muscle mass. Therefore, the purpose of this investigation was to evaluate the effects of megestrol acetate ingestion on circulating IL-15 and IL-18 concentrations in healthy elderly men. METHODS: All participants received 800 mg of megestrol acetate per day during this 12-week study. Megestrol acetate was combined with testosterone injections (100 mg/week), placebo injections, resistance training, or resistance training and testosterone. Resting IL-15 and IL-18 concentrations were measured by enzyme-linked immunosorbent assay at week 0 (pre), week 6 (mid), and week 12 (post). RESULTS: The time effect for IL-15 was significant (p = .0008), with the mid and post values being significantly greater than the pre value. The change in IL-15 concentration was not significantly related to the change in muscle mass (r = -.31; p > .05), nor was it related to the change in fat mass (r =.17; p > .05). Differences among groups or over time were not significant for IL-18, nor were correlations between pre body weight and pre IL-18 (r = -.03), pre fat mass and pre IL-18 (r = .14), or the change in fat mass and the change in IL-18 (r = -.07). CONCLUSIONS: IL-15 was increased as a result of megestrol acetate ingestion; however, megestrol acetate did not affect circulating IL-18 concentrations, and the change in IL-18 did not correlate with any body composition variables. PMID- 15345739 TI - A twin study of the genetic contribution to age-related functional impairment. AB - BACKGROUND: A key element in the quality of later life is the prevalence of age related functional impairments. The objective of this study was to quantify the genetic and environmental influences on age-related functional impairment in a population of white male twin elders who were fit in young adulthood when entering military service. The extent of genetic influence on functioning in later life affects the role of public health, personal initiative, and service interventions. METHODS: Indicators of functional impairment were determined by telephone survey and by twin pair responses to 10 indicators of basic, instrumental, and social activities, and mobility. Responses were analyzed using structural equation modeling. Prevalence and concordances were determined by zygosity status. Covariance was partitioned between twins in a pair into components attributable to additive genetics, common environment, and unique environment. RESULTS: Data from 2721 twin pairs (1384 monozygotic and 1337 dizygotic) were analyzed for the 10 dichotomous indicators of functional impairment and for a subscale of 8 of these indicators. For the subscale, additive genes accounted for approximately 21% of covariance in liability for a higher score, whereas unique environment accounted for approximately 78% of variance, with age accounting for a very small proportion. In two indicators there were nontrivial effects of common environment. CONCLUSIONS: Within the expressed limits on generalization, the study findings suggest a major potential role for interventions aimed at a person's unique environment to maintain good functioning in aging and to lengthen the period of active life. Genetic effects play a modest but also important role in age-related functional impairment. PMID- 15345740 TI - A longitudinal study of parkinsonism and disability in a community population of older people. AB - BACKGROUND: Parkinsonian signs in patients without Parkinson's disease are often undetected but occur frequently in older people, and are often considered benign. We measured the association between parkinsonism and subsequent disability. METHODS: We conducted a prospective, longitudinal, community-based population study in East Boston, Massachusetts, using a stratified random sample of 455 community residents, aged 65 years and older. Four categories of parkinsonian signs were measured at baseline: bradykinesia, gait disturbance, rigidity, and tremor. Parkinsonism was defined as the presence of two or more categories of parkinsonian signs, and cases of Parkinson's disease were excluded from analyses. Disability was assessed annually over a mean of 4.7 years using the Katz, Rosow Breslau, and Nagi disability measures. RESULTS: Parkinsonism at baseline strongly predicted subsequent disability as assessed with the three disability measures (on the Katz measure, beta = -1.30, p < .001). On average, a person with parkinsonism had a disability level at follow-up equal to that of a comparable person approximately 16.7 years for men and 8 years for women. The rate of developing disability for persons with parkinsonism increased each year; on the Katz measure, participants with parkinsonism declined on average 16.4% faster than those without. Gait impairment and bradykinesia strongly predicted subsequent disability; tremor and rigidity did not. The effect of parkinsonism was attenuated but still persistent in persons with coexistent moderate-to-severe cognitive impairment or stroke. CONCLUSIONS: Parkinsonism strongly predicts progressive disability in the older community population and has a marked aging effect on disability level. PMID- 15345741 TI - The extracellular matrix gene Frem1 is essential for the normal adhesion of the embryonic epidermis. AB - Fraser syndrome is a rare recessive disorder characterized by cryptophthalmos, syndactyly, renal defects, and a range of other developmental abnormalities. Because of their extensive phenotypic overlap, the mouse blebbing mutants have been considered models of this disorder, and the recent isolation of mutations in Fras1 in both the blebbed mouse and human Fraser patients confirms this hypothesis. Here we report the identification of mutations in an extracellular matrix gene Fras1-related extracellular matrix gene 1 (Frem1) in both the classic head blebs mutant and in an N-ethyl-N-nitrosourea-induced allele. We show that inactivation of the gene results in the formation of in utero epidermal blisters beneath the lamina densa of the basement membrane and also in renal agenesis. Frem1 is expressed widely in the developing embryo in regions of epithelial/mesenchymal interaction and epidermal remodeling. Furthermore, Frem1 appears to act as a dermal mediator of basement membrane adhesion, apparently independently of the other known "blebs" proteins Fras1 and Grip1. Unlike both Fras1 and Grip1 mutants, collagen VI and Fras1 deposition in the basement membrane is normal, indicating that the protein plays an independent role in epidermal differentiation and is required for epidermal adhesion during embryonic development. PMID- 15345742 TI - Targeted cytoplasmic irradiation induces bystander responses. AB - The observation of radiation-induced bystander responses, in which cells respond to their neighbors being irradiated, has important implications for understanding mechanisms of radiation action particularly after low-dose exposure. Much of this questions the current dogma of direct DNA damage driving response in irradiated systems. In this study, we have used a charged-particle microbeam to target individual helium ions ((3)He(2+)) to individual cells within a population of radioresistant glioma cells cultured alone or in coculture with primary human fibroblasts. We found that even when a single cell within the glioma population was precisely traversed through its cytoplasm with one (3)He(2+) ion, bystander responses were induced in the neighboring nonirradiated glioma or fibroblasts so that the yield of micronuclei was increased by 36% for the glioma population and 78% for the bystander fibroblast population. Importantly, the yield of bystander induced micronuclei was independent of whether the cytoplasm or nucleus of a cell was targeted. The bystander responses were fully eliminated when the populations were treated with 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3 oxide or filipin, which scavenge nitric oxide (NO) and disrupt membrane rafts, respectively. By using the probe 4-amino-5-methylamino-2',7'-difluorofluorescein, it was found that the NO level in the glioma population was increased by 15% after 1 or 10 cytoplasmic traversals, and this NO production was inhibited by filipin. This finding shows that direct DNA damage is not required for switching on of important cell-signaling mechanisms after low-dose irradiation and that, under these conditions, the whole cell should be considered a sensor of radiation exposure. PMID- 15345743 TI - Specialized and redundant roles of TBP and a vertebrate-specific TBP paralog in embryonic gene regulation in Xenopus. AB - The general transcription factor TATA-binding protein (TBP) is a key initiation factor involved in transcription by all three eukaryotic RNA polymerases. In addition, the related metazoan-specific TBP-like factor (TLF/TRF2) is essential for transcription of a distinct subset of genes. Here we characterize the vertebrate-specific TBP-like factor TBP2, using in vitro assays, in vivo antisense knockdown, and mRNA rescue experiments, as well as chromatin immunoprecipitation. We show that TBP2 is recruited to promoters in Xenopus oocytes in the absence of detectable TBP recruitment. Furthermore, TBP2 is essential for gastrulation and for the transcription of a subset of genes during Xenopus embryogenesis. In embryos, TBP2 protein is much less abundant than TBP, and moderate overexpression of TBP2 partially rescues an antisense knockdown of TBP levels and restores transcription of many TBP-dependent genes. TBP2 may be a TBP replacement factor in oocytes, whereas in embryos both TBP and TBP2 are required even though they exhibit partial redundancy and gene selectivity. PMID- 15345745 TI - Estrogen receptor (ER)-mediated transcriptional regulation of the human corticotropin-releasing hormone-binding protein promoter: differential effects of ERalpha and ERbeta. AB - CRH-binding protein (CRH-BP) regulates activation of the hypothalamic-pituitary adrenal (HPA) axis by binding and inhibiting CRH. We investigated for the first time transcriptional regulation of the human CRH-BP promoter using transient transfections. Estrogen receptors (ERs) contributed to ligand-independent constitutive activation of the promoter, whereas in the presence of estradiol ERalpha induced and ERbeta repressed promoter activity in a dose-dependent manner. TNFalpha inhibited promoter induction by ERalpha in the absence and presence of estradiol. Three ERE half-sites in the CRH-BP promoter bound ERalpha and ERbeta in an EMSA, and disruption of ERE half-sites by site-directed mutagenesis abolished ligand-independent induction by ERalpha and ERbeta and promoter enhancement by estradiol-activated ERalpha. Repression by estradiol/ERbeta was unaffected by disruption of ERE half-sites, activating protein 1, cAMP response element, GATA, or nuclear factor kappaB sites, and reversed to promoter induction by estrogen antagonists, tamoxifen and ICI 182,780, suggesting corepressor involvement. In hypothalamic GT1-7 cells, Western blotting demonstrated rapid induction of endogenous CRH-BP expression by estradiol-bound ER, which was inhibited by TNFalpha. We propose a model in which ERs maintain basal CRH-BP expression in pituitary and neurosecretory cells, whereas in the presence of ERalpha estrogen enhances CRH-BP transcription, causing down-regulation of the HPA axis, and nuclear factor kappaB-activating cytokines activate the HPA axis by inhibiting ERalpha. PMID- 15345744 TI - Comparative structural modeling and inference of conserved protein classes in Drosophila seminal fluid. AB - The constituents of seminal fluid are a complex mixture of proteins and other molecules, most of whose functions have yet to be determined and many of which are rapidly evolving. As a step in elucidating the roles of these proteins and exposing potential functional similarities hidden by their rapid evolution, we performed comparative structural modeling on 28 of 52 predicted seminal proteins produced in the Drosophila melanogaster male accessory gland. Each model was characterized by defining residues likely to be important for structure and function. Comparisons of known protein structures with predicted accessory gland proteins (Acps) revealed similarities undetectable by primary sequence alignments. The structures predict that Acps fall into several categories: regulators of proteolysis, lipid modifiers, immunity/protection, sperm-binding proteins, and peptide hormones. The comparative structural modeling approach indicates that major functional classes of mammalian and Drosophila seminal fluid proteins are conserved, despite differences in reproductive strategies. This is particularly striking in the face of the rapid protein sequence evolution that characterizes many reproductive proteins, including Drosophila and mammalian seminal proteins. PMID- 15345746 TI - A conformationally sensitive GHR [growth hormone (GH) receptor] antibody: impact on GH signaling and GHR proteolysis. AB - The GH receptor (GHR) mediates metabolic and somatogenic actions of GH. Its extracellular domain (ECD; residues 1-246) has two subdomains, each with seven beta strands organized into two antiparallel beta sheets, connected by a short hinge region. Most of the ECD residues involved in GH binding reside in subdomain 1, whereas subdomain 2 harbors a dimerization interface between GHR dimers that alters conformation in response to GH. A regulated GHR metalloprotease cleavage site is in the membrane-proximal stem region of subdomain 2. We have identified a monoclonal anti-ECD antibody, anti-GHR(ext-mAb), which recognizes the rabbit and human GHRs by immunoprecipitation, but less so after GH treatment. By immunoblotting and immunoprecipitation, anti-GHR(ext-mAb) recognized a glutathione-S-transferase (GST) fusion incorporating subdomain 2, but not one including subdomain 1. In transient transfection experiments, anti-GHR(ext-mAb) failed to recognize by immunoprecipitation a previously characterized dimerization interface mutant GHR that is incompetent for signaling. In signaling experiments, brief pretreatment of GH-responsive human fibrosarcoma cells with anti-GHR(ext-mAb) dramatically inhibited GH-induced Janus kinase 2 and signal transducer and activator of transcription 5 tyrosine phosphorylation and prevented GH-induced GHR disulfide linkage (a reflection of GH-induced conformational changes). In contrast, anti-GHR(ext-mAb) only partially inhibited radiolabeled GH binding, suggesting its effects on signaling were not simply via inhibition of binding. Furthermore, anti-GHR(ext-mAb) prevented phorbol ester stimulated GHR proteolysis, but GHR cleavage site mutants were normally recognized by the antibody, indicating that the stem region cleavage site is not a direct epitope. A Fab fragment of anti-GHR(ext-mAb) inhibited GH-induced GHR disulfide linkage and signaling, as well as phorbol ester-induced GHR proteolysis, in a fashion similar to the intact antibody. Thus, our findings suggest that anti-GHR(ext-mAb) has promise as a GH antagonist and as a tool in studies of conformational changes required for GHR activation. PMID- 15345747 TI - Phosphoproteomic analysis of the developing mouse brain. AB - Proper development of the mammalian brain requires the precise integration of numerous temporally and spatially regulated stimuli. Many of these signals transduce their cues via the reversible phosphorylation of downstream effector molecules. Neuronal stimuli acting in concert have the potential of generating enormous arrays of regulatory phosphoproteins. Toward the global profiling of phosphoproteins in the developing brain, we report here the use of a mass spectrometry-based methodology permitting the first proteomic-scale phosphorylation site analysis of primary animal tissue, identifying over 500 protein phosphorylation sites in the developing mouse brain. PMID- 15345748 TI - Reducing chloride conductance prevents hyperkalaemia-induced loss of twitch force in rat slow-twitch muscle. AB - Exercise-induced loss of skeletal muscle K(+) can seriously impede muscle performance through membrane depolarization. Thus far, it has been assumed that the negative equilibrium potential and large membrane conductance of Cl(-) attenuate the loss of force during hyperkalaemia. We questioned this idea because there is some evidence that Cl(-) itself can exert a depolarizing influence on membrane potential (V(m)). With this study we tried to identify the possible roles played by Cl(-) during hyperkalaemia. Isolated rat soleus muscles were kept at 25 degrees C and twitch contractions were evoked by current pulses. Reducing [Cl(-)](o) to 5 mM, prior to introducing 12.5 mM K(o), prevented the otherwise occurring loss of force. Reversing the order of introducing these two solutions revealed an additional effect, i.e. the ongoing hyperkalaemia-related loss of force was sped up tenfold after reducing [Cl(-)](o). However, hereafter twitch force recovered completely. The recovery of force was absent at [K(+)](o) exceeding 14 mM. In addition, reducing [Cl(-)](o) increased membrane excitability by 24%, as shown by a shift in the relationship between force and current level. Measurements of V(m) indicated that the antagonistic effect of reducing [Cl( )](o) on hyperkalaemia-induced loss of force was due to low-Cl(-)-induced membrane hyperpolarization. The involvement of specific Cl(-) conductance was established with 9-anthracene carboxylic acid (9-AC). At 100 microm, 9-AC reduced the loss of force due to hyperkalaemia, while at 200 microm, 9-AC completely prevented loss of force. To study the role of the Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) in this matter, we added 400 microm of the NKCC inhibitor bumetanide to the incubation medium. This did not affect the hyperkalaemia-induced loss of force. We conclude that Cl(-) exerts a permanent depolarizing influence on V(m). This influence of Cl(-) on V(m), in combination with a large membrane conductance, can apparently have two different effects on hyperkalaemia-induced loss of force. It might exert a stabilizing influence on force production during short periods of hyperkalaemia, but it can add to the loss of force during prolonged periods of hyperkalaemia. PMID- 15345749 TI - Transcellular thiocyanate transport by human airway epithelia. AB - Human airway mucosa synthesizes and secretes lactoperoxidase (LPO). As H(2)O(2) and thiocyanate (SCN(-)) are also present, a functional LPO antibacterial defence system exists in the airways. SCN(-) concentrations in several epithelial secretions are higher than in serum, although the mechanisms of transepithelial transport and accumulation in these secretions are unknown. To examine SCN(-) accumulation in secretions, human airway epithelial cells, re-differentiated at the air-liquid interface, were used in open-circuit conditions. [(14)C]SCN(-), in the basolateral medium, was transported across the epithelium and concentrated tenfold at the apical surface. Measurement of the transepithelial potential showed that the basolateral compartment was positive relative to the apical surface (13.7 +/- 1.8 mV) and therefore unfavourable for passive movement of SCN( ). Transport was dependent on basolateral [SCN(-)] and saturable (K(m,app) = 69 +/- 25 microM); was inhibited by increased apical [SCN(-)]; and was dependent on the presence of basolateral Na(+). Perchlorate (K(i,app) = 0.6 +/- 0.05 microM) and iodide (K(i,app) = 9 +/- 8 microM) in the basolateral medium reversibly inhibited transport, but furosemide did not. Iodide was also transported (K(m,app) = 111 +/- 69 microM). RT-PCR and immunohistochemistry confirmed expression of Na(+)-I(-) symporter (NIS) in the airways. SCN(-) transport was insensitive to apical disulphonic acid Cl(-) channel blockers, but sensitive to apical glibenclamide and arylaminobenzoates. Forskolin and dibutyryl cAMP increased transport. These data suggest SCN(-) transport may occur through basolateral NIS-mediated SCN(-) concentration inside cells, followed by release through an apical channel, perhaps cystic fibrosis transmembrane conductance regulator. PMID- 15345750 TI - Inhibition of KATP channel activity augments baroreflex-mediated vasoconstriction in exercising human skeletal muscle. AB - In the present investigation we examined the role of ATP-sensitive potassium (K(ATP)) channel activity in modulating carotid baroreflex (CBR)-induced vasoconstriction in the vasculature of the leg. The CBR control of mean arterial pressure (MAP) and leg vascular conductance (LVC) was determined in seven subjects (25 +/- 1 years, mean +/- S.E.M.) using the variable-pressure neck collar technique at rest and during one-legged knee extension exercise. The oral ingestion of glyburide (5 mg) did not change mean arterial pressure (MAP) at rest (86 versus 89 mmHg, P > 0.05), but did appear to increase MAP during exercise (87 versus 92 mmHg, P = 0.053). However, the CBR-MAP function curves were similar at rest before and after glyburide ingestion. The CBR-mediated decrease in LVC observed at rest (approximately 39%) was attenuated during exercise in the exercising leg (approximately 15%, P < 0.05). Oral glyburide ingestion partially restored CBR-mediated vasoconstriction in the exercising leg (approximately 40% restoration, P < 0.05) compared to control exercise. These findings indicate that K(ATP) channel activity modulates sympathetic vasoconstriction in humans and may prove to be an important mechanism by which functional sympatholysis operates in humans during exercise. PMID- 15345751 TI - Local protein synthesis and GABAB receptors regulate the reversibility of long term potentiation at murine hippocampal mossy fibre-CA3 synapses. AB - Reversal of long-term potentiation (LTP) by long trains of low-frequency stimulation is generally referred to as depotentiation. One of the intriguing aspects of depotentiation is that the magnitude of depotentiation is inversely proportional to the time lag of depotentiation stimulation following LTP induction. Although the mechanisms underlying depotentiation have been widely explored, the factors that regulate the susceptibility of LTP to depotentiation stimulation remain largely unclear. We now report that multiple trains of high frequency stimulation provide immediate synaptic resistance to depotentiation stimulation at the mossy fibre-CA3 synapses. The synaptic resistance to depotentiation stimulation depends on the amount of synaptic stimulation used to induce LTP; it is prevented by protein synthesis inhibitors and is input specific. In contrast, neither the transection of mossy fibre axons near granule cell somata nor the application of RNA synthesis inhibitors influences synaptic resistance to depotentiation stimulation. We also provide evidence that the induction of depotentiation is regulated by GABA(B) receptors. Application of a GABA(B) receptor antagonist significantly promoted the synaptic resistance to depotentiation stimulation, whereas inhibition of GABA transport delayed the onset of this synaptic resistance. These results suggest that local protein synthesis is required for the development of synaptic resistance to depotentiation stimulation, whereas the activation of GABA(B) receptors promotes the susceptibility to depotentiation stimulation. These two factors may crucially regulate the reversal and stability of long-term information storage. PMID- 15345752 TI - Induction of novel agonist selectivity for the ADP-activated P2Y1 receptor versus the ADP-activated P2Y12 and P2Y13 receptors by conformational constraint of an ADP analog. AB - ADP is the cognate agonist of the P2Y1, P2Y12, and P2Y13 receptors. With the goal of identifying a high potency agonist that selectively activates the P2Y1 receptor, we examined the pharmacological selectivity of the conformationally constrained non-nucleotide analog (N)-methanocarba-2MeSADP [(1'S,2'R, 3'S,4'R,5'S)-4-[(6-amino-2-methylthio-9H-purin-9-yl)-1 diphosphoryloxymethyl]bicyclo[3.1.0]hexane-2,3-diol] among the three ADP activated receptors. Each P2Y receptor was expressed transiently in COS-7 cells, and inositol lipid hydrolysis was quantified as a measure of receptor activity. In the case of the Gi-linked P2Y12 and P2Y13 receptors, a chimeric G protein, Galphaq/i, was coexpressed to confer a capacity of these Gi-linked receptors to activate phospholipase C. 2MeSADP (2-methylthio-ADP) was a potent agonist at all three receptors exhibiting EC50 values in the sub to low nanomolar range. In contrast, whereas (N)-methanocarba-2MeSADP was an extremely potent (EC50=1.2 +/- 0.2 nM) agonist at the P2Y1 receptor, this non-nucleotide analog exhibited no agonist activity at the P2Y12 receptor and very low activity at the P2Y13 receptor. (N)-Methanocarba-2MeSADP also failed to block the action of 2MeSADP at the P2Y12 and P2Y13 receptors, indicating that the (N)-methanocarba analog is not an antagonist at these receptors. The P2Y1 receptor selectivity of (N) methanocarba-2MeSADP was confirmed in human platelets where it induced the shape change promoted by P2Y1 receptor activation without inducing the sustained platelet aggregation that requires simultaneous activation of the P2Y12 receptor. These results provide the first demonstration of a high-affinity agonist that discriminates among the three ADP-activated P2Y receptors, and therefore, introduce a potentially important new pharmacological tool for delineation of the relative biological action of these three signaling proteins. PMID- 15345753 TI - Calcium-activated potassium channel triggers cardioprotection of ischemic preconditioning. AB - We tested the hypothesis that the high-conductance calciumactivated potassium (K(Ca)) channel is involved in the cardioprotection of preconditioning with ischemic insults. In the isolated perfused rat heart subjected to ischemia/reperfusion, effects of ischemic preconditioning (IPC) on infarct size and lactate dehydrogenase (LDH) release were abolished by 1 microM paxilline (Pax), an inhibitor of the K(Ca) channel, administered 30 min before, but not during, ischemia. In isolated ventricular myocytes subjected to metabolic inhibition and anoxia (MI/A), preconditioning with MI/A increased their viability, and the effect was abolished by administering Pax before MI/A. Like IPC, 10 microM NS1619 (1,3-dihydro-1-[2-hydroxy-5-(trifluoromethyl)phenyl]-5 trifluoromethyl-2Hbenzimidazol-2-one; NS), an opener of K(Ca) channels, reduced infarct size and LDH release, effects attenuated by Pax. The harmful and protective effects of blockade and activation of the K(Ca) channel were accompanied by impaired and improved left ventricular contractile functions, respectively. In addition, the effect of NS was not altered by 100 microM 5 hydroxydecanoate, an inhibitor of the K(ATP) channel. Neither was the effect of 100 microM diazoxide, an activator of the K(ATP) channel, altered by Pax. Furthermore, opening of the mitochondrial permeability transition pore (mPTP) with 20 microM atractyloside abolished the beneficial effects of IPC or NS in the isolated rat heart and myocyte. Inhibition of mPTP opening with 0.2 microM cyclosporin A decreased the infarct size and LDH release and improved the contractile function, effects not attenuated by Pax. In conclusion, the study provides evidence that the K(Ca) channel triggers cardioprotection of IPC, which involves mPTP. PMID- 15345754 TI - Health care and the homeless. PMID- 15345755 TI - State mental health policy: Another chance to do it right: redesigning public behavioral health care in New Mexico. PMID- 15345756 TI - Datapoints: Service availability for persons with co-occurring conditions. PMID- 15345757 TI - State mental health policy: A unique approach to designing a comprehensive behavioral health system in New Mexico. PMID- 15345758 TI - Rehab rounds: Cultural adaptation of the basic conversational skills module for a Chinese population. PMID- 15345759 TI - Practical application of pharmacotherapy with long-acting risperidone for patients with schizophrenia. AB - OBJECTIVE: It is now generally accepted that the use of second-generation, or atypical, antipsychotics for schizophrenia represents an advance over conventional antipsychotic agents. However, adherence continues to be a problem, as with other medications for chronic disorders. Long-acting formulations of conventional antipsychotics partly address adherence problems, but their use is limited by tolerability issues. This article provides practical advice to physicians on the characteristics of patients who would benefit from treatment with long-acting atypical antipsychotic agents and offers suggestions on how to initiate treatment. METHODS: A literature search for studies published between 1980 and 2003 that evaluated the treatment of patients with schizophrenia with long-acting atypical agents was conducted by using MEDLINE and EMBASE. The primary search parameters were "schizophrenia," "atypical," "antipsychotic," and "long-acting." As expected, long-acting risperidone was the only long-acting atypical agent identified; thus this article focuses on practical advice and suggestions on how to initiate therapy with long-acting risperidone. RESULTS AND DISCUSSION: From the results of the literature search and the discussion of a panel of experts at a meeting held in Dublin in 2003 and supported by Johnson & Johnson, it is possible to conclude that long-acting risperidone has demonstrated efficacy and tolerability, even among patients who are considered clinically stable on other antipsychotics. Most patients can switch safely and effectively to long-acting risperidone if appropriate strategies are applied. Long-acting risperidone provides a new and promising therapeutic option for the treatment of schizophrenia. PMID- 15345760 TI - Relationship between antipsychotic medication treatment and new cases of diabetes among psychiatric inpatients. AB - OBJECTIVE: This study examined data on patients with serious and persistent mental illness in a large state hospital system to determine whether patients who took second-generation antipsychotics were more likely to develop diabetes mellitus than patients who took first-generation antipsychotics. METHODS: A case control study design was used. A new prescription of an antidiabetic medication was used to identify new cases of diabetes mellitus. Odds ratios were calculated for exposure to second-generation antipsychotics (clozapine, risperidone, olanzapine, quetiapine, and multiple second-generation antipsychotics) compared with exposure to first-generation antipsychotics. Cases and controls were identified by using a database that contained drug prescription information from the inpatient facilities that were operated by the New York State Office of Mental Health. Data from January 1, 2000, to December 31, 2002, were examined. Among 13,611 unique patients who received antipsychotics, 8,461 met entry criteria of being hospitalized for at least 60 days and not having an antidiabetic medication prescribed in the past. A total of 181 of these inpatients received prescriptions for an antidiabetic medication at least 30 days after their admission. Eight controls (N=1,448) for each case (N=181) were matched by calendar year, length of observation period, race, age group, and diagnosis, giving a total sample of 1,629 patients. RESULTS: Statistically significant elevations in risk were seen among patients who received more than one second-generation antipsychotic or clozapine or quetiapine, compared with patients who received first-generation antipsychotics alone. Although not statistically significant, odds ratios for olanzapine and risperidone were also elevated. Conditional logistic regression adjusting for gender and age did not change the results. CONCLUSIONS: Exposure to multiple second-generation antipsychotics or clozapine or quetiapine significantly increased the risk of treatment-emergent diabetes mellitus. PMID- 15345761 TI - Age-related modifiers of clinical presentation and health service use among veterans with bipolar disorder. AB - OBJECTIVE: This retrospective analysis of a large Department of Veterans Affairs (VA) database evaluated differences in clinical presentation, health service use, and use of psychotropic medications between older and younger adults with bipolar disorder. METHODS: The results presented in this article focus on veterans with bipolar disorder who were active in the VA's National Psychosis Registry in federal fiscal year 2001 (FY01). RESULTS: The registry listed 65,556 individuals as having bipolar disorder in FY01. In FY01, nearly 25 percent of veterans with bipolar disorder (more than 16,000 individuals) were aged 60 years or older, and more than 10 percent were aged 70 years or older. Elderly persons were hospitalized at similar or slightly lower rates than middle-aged persons but tended to have longer hospital stays. Use of outpatient services was also higher in older populations. Patterns of use of psychotropic medications indicated that valproate is the most commonly used mood-stabilizing agent in the VA, in use by 72.9 percent of patients who receive a mood stabilizer. Use of lithium is substantial as well, with more than 40 percent of patients taking this agent. In addition, just over 40 percent of all veterans with bipolar disorder receive antipsychotic medication. CONCLUSIONS: Bipolar disorder affects large numbers of veterans across the entire life span, and use of VA resources by this population becomes greater with advancing age. Data from this study provide new information about the unique treatment requirements and clinical presentations of older patients with bipolar disorder. PMID- 15345762 TI - Treatment retention among children entering a new episode of mental health care. AB - OBJECTIVE: This study examined use of mental health services among children and adolescents with private insurance who were entering treatment. Variations in service use were examined by age, gender, diagnosis, recent psychiatric hospitalization, and type of insurance. Differences between children who received treatment from mental health professionals and those who were treated by primary care physicians were also examined. METHODS: Drawn from a large database, the sample comprised 11,659 new users of mental health services. Service use was defined as the total number of days children were retained in treatment and the total number of mental health contacts recorded. RESULTS: The overall mean number of visits within a six-month period was 3.9. The average duration of treatment was 75.36 days. Children who were treated by a mental health specialist were less likely to drop out of treatment and had a larger number of visits. Severity of illness, psychiatric hospitalization, and managed care insurance coverage were also associated with lower risk of dropout and greater intensity of care. CONCLUSIONS: Children's access to services does not guarantee sustained involvement in treatment. To more fully address the nature of service use among children, a closer look at specific barriers to continued involvement in services is needed. PMID- 15345763 TI - Impact of family burden and affective response on clinical outcome among patients with bipolar disorder. AB - OBJECTIVE: This study evaluated the direct and indirect effects of family burden and affective response on medication adherence and outcome among patients with bipolar disorder. METHODS: Data were examined for 126 patients who were consecutively admitted to the psychiatric service at a university-affiliated hospital and who met research diagnostic criteria for bipolar I or II disorder or for schizoaffective disorder, manic type, and their family caregivers. A total of 101 pairs of patients and family caregivers (80 percent) completed 15 months of study and were included in the analyses. Patients and their identified caregivers were assessed within two weeks of either discharge from the index inpatient admission or initiation of outpatient treatment (baseline assessment). Patients and caregivers were also assessed seven and 15 months after the baseline assessment. Structural equation modeling was used to evaluate caregivers' influences on patients' medication adherence seven months after baseline and on clinical outcome 15 months after baseline. RESULTS: The indexes of overall fit for the path model confirmed the a priori measurement model. Significant paths were found from the caregiver's perceived burden at baseline to the caregiver's emotional overinvolvement at baseline, from the caregiver's emotional overinvolvement at baseline to the patient's medication adherence at the seven month follow-up, and from the patient's medication adherence at the seven-month follow-up to the patient's outcome at the 15-month follow-up. The paths from the caregiver's perceived burden at baseline to the patient's medication adherence seven months after baseline and the patient's outcome 15 months after baseline were not significant. CONCLUSIONS: When caregivers of patients with bipolar illness experience a high burden, patient outcome is adversely affected. This relationship is mediated through families' affective response and patients' medication adherence. PMID- 15345764 TI - Mental health issues among female clients of domestic violence programs in North Carolina. AB - OBJECTIVE: This study estimated the prevalence of mental health problems among clients of domestic violence programs in North Carolina, determined whether domestic violence program staff members routinely screen clients for mental health problems, described how domestic violence programs respond to clients who have mental health problems, and ascertained whether domestic violence program staff members and volunteers have been trained in mental health-related issues. METHODS: A survey was mailed to all known domestic violence programs in North Carolina. RESULTS: A total of 71 of the 84 known programs responded to the survey (85 percent response rate). A majority of programs estimated that at least 25 percent of their clients had mental health problems (61 percent) and stated that they routinely asked their clients about mental health issues (72 percent). More than half the programs (54 percent) reported that less than 25 percent of their staff members and volunteers had formal training on mental health issues. An even smaller percentage of programs (23 percent) reported that they had a memorandum of agreement with a local mental health center. CONCLUSIONS: The substantial percentage of domestic violence clients with concurrent mental health needs and the limited mental health services that are currently available have important implications for domestic violence and mental health service delivery. PMID- 15345765 TI - Multimodal treatment for ADHD among youths in three Medicaid subgroups: disabled, foster care, and low income. AB - OBJECTIVE: This study compared the use of treatments for attention-deficit hyperactivity disorder (ADHD) among three distinct subpopulations of Medicaid insured youths who have very different mental health needs and patterns of service use: those with federally documented disability, those in foster care, and those in families with low income. METHODS: This one-year, cross-sectional study of community mental health services used administrative data. Individuals who were younger than 20 years, who were continuously enrolled in one Mid Atlantic state Medicaid program, and who had two or more medical encounters associated with an ADHD diagnosis in 1998 were identified (N=1,296). Measures of the use of mental health services were the number of different classes of psychopharmacologic medications, the psychopharmacologic regimen, and the combined use of pharmacotherapy and psychotherapy treatments (multimodal treatment). RESULTS: Use of multiple psychopharmacologic agents was greater in the disabled and foster care groups compared with the low-income group. Significantly fewer mental health provider visits, but greater use of stimulant treatment only, were observed in the low-income group compared with the other groups. Youths in the disabled group were significantly more likely than youths in the low-income group, but not more likely than youths in the foster care group, to receive multimodal treatments. Children in foster care were significantly more likely than those in the other groups to use a substance abuse service. CONCLUSIONS: Among a cohort of Medicaid-enrolled youths with ADHD, co existing psychiatric disorders and complex psychopharmacologic treatments were more common in the disabled and foster care groups than in the low-income group. Youths with disabilities were significantly more likely than youths in the low income group to receive multimodal treatment. PMID- 15345766 TI - Comparative prevalence of psychotropic medications among youths enrolled in the SCHIP and privately insured youths. AB - The one-year prevalence of use of psychotropic medications among youths enrolled in a Mid-Atlantic State Children's Insurance Program (SCHIP) in 1999 or 2000 was compared with the 1998-2000 prevalence findings for youths enrolled in four large multistate private medical insurance programs. Prevalence was significantly higher among youths enrolled in the SCHIP than among those with private insurance in all eight medication group comparisons. Unlike private insurance, the surveyed state SCHIP program did not require a copayment or have a restrictive formulary. These administrative differences may partly explain the comparatively higher rate of psychotropic use under the SCHIP. PMID- 15345767 TI - The independent living scales as a measure of functional outcome for schizophrenia. AB - The Independent Living Scales (ILS) measures cognitive skills required for independent living and is intended to provide guidelines for appropriate supervision requirements for persons in residential placement. To assess the validity of the ILS among persons with schizophrenia, the instrument was administered to 162 individuals with schizophrenia who were living in three gradations of care: maximum supervision, moderate supervision, and minimal supervision. Scores on the ILS differed significantly across the three levels of care, whereas scores on the Global Assessment of Functioning (GAF) that were provided by clinicians discriminated only two levels of care. The ILS can be used among patients with schizophrenia to measure cognition as it affects functional outcome. PMID- 15345768 TI - Survey of stress reactions among health care workers involved with the SARS outbreak. AB - The outbreak of severe acute respiratory syndrome (SARS) was unique because it was highly concentrated in health care settings and a large number of health care workers were infected. This study investigated stress reactions among 338 staff members in a hospital in East Taiwan that discontinued emergency and outpatient services to prevent possible nosocomial outbreak. Seventeen staff members (5 percent) suffered from an acute stress disorder; stepwise multiple logistic regression analysis determined that quarantine was the most related factor. Sixty six staff members (20 percent) felt stigmatized and rejected in their neighborhood because of their hospital work, and 20 of 218 health care workers (9 percent) reported reluctance to work or had considered resignation. PMID- 15345769 TI - Psychiatrists' attitudes toward involuntary hospitalization. AB - This study examined whether psychiatrists' attributions of responsibility for mental illnesses affect their decisions about involuntary hospitalization. A survey that was mailed in 2002 to members of the Illinois Psychiatric Society elicited recommendations for involuntary commitment for vignette characters. The survey also sought respondents' attributions of personal responsibility for the onset and recurrence of mental illnesses. A total of 432 psychiatrists responded to the survey. Decisions to involuntarily hospitalize persons with mental illness increased significantly with the level of risk of harm and varied significantly between psychiatric diagnoses. Attributions of responsibility were not related to commitment decisions. PMID- 15345770 TI - Posttraumatic stress disorder, depression, and perceived safety 13 months after September 11. AB - This study assessed relationships between exposure to the September 11, 2001, terrorist attack, current posttraumatic stress disorder (PTSD), current major depression, and current safety perceptions in a sample of 212 Pentagon staff members 13 months after the attack. Forty-eight respondents (23 percent) had possible PTSD; eight (4 percent) had probable major depression. Respondents who were directly exposed to the attack were more likely to have PTSD and major depression and were less likely to have a perception of safety at work and in usual activities and travel only. In contrast, respondents with PTSD reported a lower perception of safety at home, at work, and in usual activities and travel. PMID- 15345771 TI - A case series of nicotine nasal spray in the treatment of tobacco dependence among patients with schizophrenia. AB - A retrospective case series of 12 smokers with schizophrenia or schizoaffective disorder who had not successfully quit smoking with previous treatments for tobacco dependence were treated with nicotine nasal spray. All but one patient (92 percent) tolerated the nasal spray well, and nine (75 percent) used it at maximal doses for prolonged periods. After treatment five patients (42 percent) were abstinent from smoking for more than 90 days, and four patients (33 percent) substantially reduced the amount that they smoked. Ten patients (83 percent) used the spray in combination with other medications, and all received psychosocial support. Nicotine nasal spray was found to be well tolerated. PMID- 15345772 TI - Hargrave v. Vermont and the quality of care. PMID- 15345774 TI - Antihostility effects of adjunctive divalproex. PMID- 15345776 TI - Positive experiences with giving gifts to patients. PMID- 15345777 TI - Human PAD4 regulates histone arginine methylation levels via demethylimination. AB - Methylation of arginine (Arg) and lysine residues in histones has been correlated with epigenetic forms of gene regulation. Although histone methyltransferases are known, enzymes that demethylate histones have not been identified. Here, we demonstrate that human peptidylarginine deiminase 4 (PAD4) regulates histone Arg methylation by converting methyl-Arg to citrulline and releasing methylamine. PAD4 targets multiple sites in histones H3 and H4, including those sites methylated by coactivators CARM1 (H3 Arg17) and PRMT1 (H4 Arg3). A decrease of histone Arg methylation, with a concomitant increase of citrullination, requires PAD4 activity in human HL-60 granulocytes. Moreover, PAD4 activity is linked with the transcriptional regulation of estrogen-responsive genes in MCF-7 cells. These data suggest that PAD4 mediates gene expression by regulating Arg methylation and citrullination in histones. PMID- 15345778 TI - Observation of superflow in solid helium. AB - We report on the observation of nonclassical rotational inertia in solid helium-4 confined to an annular channel in a sample cell under torsional motion, demonstrating superfluid behavior. The effect shows up as a drop in the resonant oscillation period as the sample cell is cooled below 230 millikelvin. Measurement of 17 solid samples allows us to map out the boundary of this superfluid-like solid or supersolid phase from the melting line up to 66 bars. This experiment indicates that superfluid behavior is found in all three phases of matter. PMID- 15345779 TI - Avian H5N1 influenza in cats. AB - During the 2003 to 2004 outbreak of avian influenza A (H5N1) virus in Asia, there were anecdotal reports of fatal infection in domestic cats, although this species is considered resistant to influenza. We experimentally inoculated cats with H5N1 virus intratracheally and by feeding them virus-infected chickens. The cats excreted virus, developed severe diffuse alveolar damage, and transmitted virus to sentinel cats. These results show that domestic cats are at risk of disease or death from H5N1 virus, can be infected by horizontal transmission, and may play a role in the epidemiology of this virus. PMID- 15345780 TI - Physics. Superfluidity in a crystal? PMID- 15345781 TI - Tricyclic antidepressants, QT interval prolongation, and torsade de pointes. AB - The authors postulate mechanisms linking tricyclic antidepressants, QT interval prolongation, torsade de pointes, and sudden cardiac death. Case reports identify amitriptyline and maprotiline as the tricyclic antidepressants most likely to provoke torsade de pointes. Risk factors of family history of congenital long QT syndrome, age, female sex, metabolic and cardiovascular disease, metabolic inhibitors, hypokalemia, drug overdose, and co-prescription of drugs associated with QT interval prolongation were found in cases of torsade de pointes associated with tricyclic antidepressants. Clinicians should be cautious when prescribing tricyclic antidepressants with other drugs, such as thioridazine, that are capable of prolonging the QT interval. PMID- 15345782 TI - Predictors of 1-year survival assessed at the time of bone marrow transplantation. AB - The purpose of this prospective cohort study was to identify pretransplant and transplant predictors of 1-year survival after allogeneic hematopoietic stem cell transplantation (HSCT) for chronic myelogenous leukemia. Psychosocial and behavioral variables such as alcohol abuse and cigarette smoking were systematically assessed at the time of HSCT with structured diagnostic instruments. A total of 114 patients participated, with an overall 1-year survival rate of 66%. Lifetime alcohol and other substance use, cigarette smoking, depression, and quality of life prior to transplant were not found to affect 1-year survival. However, other clinical variables prior to transplant and once transplant occurred were found to predict survival. PMID- 15345783 TI - Interrater reliability, prevalence, and relation to ICD-10 diagnoses of the Diagnostic Criteria for Psychosomatic Research in consultation-liaison psychiatry patients. AB - The Diagnostic Criteria for Psychosomatic Research (DCPR) have been proposed by an international group of psychosomatic investigators as an operationalized tool for the assessment of psychological distress in medical patients. The aims of the present study were to evaluate interrater reliability, the distribution of DCPR syndromes, and their relationship with ICD-10 diagnostic categories. One hundred consecutive patients who were referred for psychiatric consultation in a university general hospital consented to assessment for DCPR syndromes as elicited in a joint interview conducted by two researchers. The results showed excellent interrater agreement, with kappa values for the 11 DCPR syndromes ranging from 0.69 to 0.97. More patients met criteria for one or more of the DCPR (87%) than for an ICD-10 diagnosis (75%). Four DCPR syndromes were particularly prevalent: demoralization, alexithymia, illness denial, and type A behavior. DCPR criteria appear to be a useful, reliable, and promising approach in the assessment and description of psychological distress in medical patients. They may serve as a focus of intervention studies in this population. PMID- 15345784 TI - Depressive symptoms, neurocognitive impairment, and adherence to highly active antiretroviral therapy among HIV-infected persons. AB - The association of depressive symptoms, neurocognitive impairment, and adherence to highly active antiretroviral therapy (HAART) was evaluated in 135 HIV-infected persons. Thirty percent reported nonadherence to HAART. Depressive symptoms (assessed with the Montgomery-Asberg Depression Rating Scale) and neurocognitive impairment (assessed with a neuropsychological test battery) were documented in 24% and 12%, respectively, of the study participants. Nonadherence to HAART was independently associated with worse depression rating scale scores (odds ratio=1.05, 95% confidence interval [CI]=1.00-1.10), acquisition of HIV through injection of drugs (odds ratio=2.59, 95% CI=1.05-6.39), and complaints about impairment of sexual activity (odds ratio=6.62, 95% CI=1.16-37.6). The presence of depressive symptoms, but not neurocognitive impairment, was associated with nonadherence. PMID- 15345785 TI - Menopausal symptoms and psychological distress in women with and without tubal sterilization. AB - Data from self-administered mailed questionnaires were used to examine the relationship between menopausal and psychological/psychiatric factors in women age 45-50 years with and without tubal sterilization. In multivariate logistic regression analysis, tubal sterilization was associated with current flushing (odds ratio = 8.78, 95% confidence interval [CI] = 1.58-48.67) and current symptoms of psychological distress (odds ratio = 3.37, 95% CI = 1.27-8.95); psychological distress was associated with tubal sterilization (odds ratio = 3.28, 95% CI = 1.28-8.42) and with being perimenopausal (odds ratio = 3.93, 95% CI = 1.08-14.34). Sterilization was marginally associated with a history of physician-diagnosed depression (univariate age-adjusted odds ratio = 2.24, 95% CI = 0.90-5.60). The menopausal and psychological symptoms of women with tubal sterilization should be taken seriously and treated appropriately. PMID- 15345786 TI - Assessment of vital exhaustion and identification of subjects at increased risk of myocardial infarction in general practice. AB - Vital exhaustion, a state characterized by unusual fatigue, loss of energy, increased irritability, and feelings of demoralization, is one of the cardiovascular risk factors. The authors investigated whether vital exhaustion contributes to the identification of subjects at increased risk of myocardial infarction in general practice. In this prospective cohort study, vital exhaustion was assessed with the Maastricht Interview on Vital Exhaustion. Other cardiovascular risk factors established were age, gender, systolic and diastolic blood pressure, total cholesterol, body mass index, smoking habits, cardiovascular disease, and diabetes mellitus. A Cox regression analysis was used. The subjects were adults (41-66 years) in an average Dutch village population. Outcome measures were fatal and nonfatal myocardial infarction. At the univariate level, vital exhaustion doubled the risk of myocardial infarction. The effect of exhaustion was confounded by gender; women had higher exhaustion scores and a lower incidence of myocardial infarction. With control for gender, age, systolic blood pressure, total cholesterol, smoking habits, self-reported cardiovascular disease, and diabetes mellitus, vital exhaustion almost tripled the risk of myocardial infarction. Assessment of vital exhaustion contributes to the identification of subjects at increased risk of myocardial infarction in general practice. PMID- 15345787 TI - Comorbid medical illness and relapse of major depressive disorder in the continuation phase of treatment. AB - The authors examined the impact of comorbid medical illness on the rate of relapse of major depressive disorder during continuation therapy. Subjects (N = 128) with major depressive disorder (according to DSM-III-R criteria) achieved clinical remission (a 17-item Hamilton Depression Rating Scale score < or = 7) after 8 weeks of treatment with fluoxetine and entered the continuation phase of antidepressant treatment. They used the Cumulative Illness Rating Scale to measure the severity of comorbid medical illness. Eight patients (6.3%) relapsed during the 28-week continuation phase. With logistic regression, the total burden and the severity of comorbid medical illness significantly predicted the relapse of major depressive disorder during continuation therapy with fluoxetine. Greater medical comorbidity was also associated with higher increases in self-reported symptoms of depression, anxiety, and anger during the follow-up. PMID- 15345788 TI - Instrument for detection of delirium in general hospitals: adaptation of the confusion assessment method. AB - Delirium is a common and severe disorder that is often misdiagnosed. The use of screening instruments is advisable for its early detection and treatment. In this study, the authors present an adaptation of the Confusion Assessment Method in order to improve its psychometric properties. One hundred fifty-three elderly inpatients were assessed in a four-phase procedure. Interrater reliability was high (kappa = 0.89). Sensitivity was 90%, and specificity was 100%; the value for negative predictive accuracy was 97%, and the value for positive predictive accuracy was 100%. The adaptation has convergent agreement with two other mental status tests, the Mini-Mental Status Examination and the Delirium Rating Scale. Our results suggest that the adaptation of the Confusion Assessment Method is sensitive, specific, reliable, and easy to use by clinicians. PMID- 15345789 TI - Is joint hypermobility related to anxiety in a nonclinical population also? AB - This study examines the association between joint hypermobility syndrome and anxiety in a nonclinical sample. Subjects (N = 526) receiving a medical check-up were assessed with the Hospital del Mar hypermobility criteria and the State Trait Anxiety Inventory. Scores for trait anxiety, and to a lesser extent state anxiety, were significantly higher among subjects with joint hypermobility syndrome than among subjects without this syndrome (median trait anxiety scores for women: 17 versus 11; median scores for men: 13 versus 1). These findings indicate that the association of joint hypermobility syndrome and anxiety holds even for subjects with no psychiatric diagnosis. Therefore, it seems that this benign connective tissue disorder is a predisposing factor for trait anxiety. However, it is necessary to further explore and define the biological basis of this syndrome, as well as its associations and clinical expressions, which interact with great complexity. PMID- 15345790 TI - Change in psychosocial functioning and quality of life of patients with body dysmorphic disorder treated with fluoxetine: a placebo-controlled study. AB - In a 12-week placebo-controlled study of fluoxetine in the treatment of body dysmorphic disorder, the authors investigated change in psychosocial functioning and mental health-related quality of life in 60 subjects. The subjects were assessed with the LIFE-RIFT (a measure of impaired functioning), Social and Occupational Functioning Scale (SOFAS), and Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) before and after receiving fluoxetine or placebo. At baseline, the patients had impaired psychosocial functioning and markedly poor mental health-related quality of life. Compared to placebo, fluoxetine was associated with significantly greater improvement in LIFE-RIFT and SOFAS scores and with improvement on the mental health subscale of the SF-36 that approached significance. Decrease in the severity of body dysmorphic disorder, as measured by the Yale-Brown Obsessive Compulsive Scale Modified for Body Dysmorphic Disorder, was significantly correlated with improvement in functioning and quality of life. PMID- 15345791 TI - Depression, estrogen, and the Women's Health Initiative. AB - This clinical observation report compares hormone use and clinical presentation in a series of middle-aged depressed women before and after publication of the Women's Health Initiative. Depressed women over age 40 seen at a general hospital academic women's affective disorders practice 6 months before and after publication of the Women's Health Initiative were compared for medication changes, hormone therapy, lifetime depressive episodes, time since last episode, time to depression recurrence after hormone cessation, symptoms, and treatment response. More women stopped hormone therapy and reported onset of depression within 3 weeks of hormone discontinuation after than before publication of the Women's Health Initiative. Depression in most women responded to reinstitution of estrogen or initiation or increase in antidepressant dose. Discontinuation of hormone therapy appears to be associated with the rapid recurrence of depression in some women with a history of depression. Randomized controlled trials in middle-aged depressed women of estrogen or a selective estrogen receptor modulator as monotherapy or as an augmentation agent are urgently needed. PMID- 15345792 TI - Does "fight or flight" need updating? PMID- 15345793 TI - Hypertensive urgency with clonidine and mirtazepine. PMID- 15345794 TI - Torsades de pointes caused by a small dose of risperidone in a terminally ill cancer patient. PMID- 15345795 TI - Apolipoprotein B/apolipoprotein A-I in relation to the metabolic syndrome and change in carotid artery intima-media thickness during 3 years in middle-aged men. AB - BACKGROUND AND PURPOSE: The apolipoprotein B (apoB)/apolipoprotein A-I (apoA-I) ratio is a measure of the relationship between different lipoprotein particles and a powerful predictor of coronary death. The aim was to examine whether apoB/apoA-I was associated with the metabolic syndrome (MetS) at baseline and also with the future change in carotid artery intima-media thickness (IMT). METHODS: In 313 58-year-old men, carotid artery IMT was measured bilaterally by high-resolution B-mode ultrasound at baseline and after 3 years of follow-up. Serum apolipoprotein concentrations and the components of MetS were measured at study entry. RESULTS: ApoB/apoA-I showed statistically significant associations with body mass index, waist-to-hip ratio, high-density lipoprotein (HDL) cholesterol, triglycerides, low-density lipoprotein (LDL) particle size, insulin, and diastolic blood pressure. Two thirds of the patients with MetS had high apoB/apoA-I ratios (>0.90) compared with one third of those without the syndrome (P<0.001). The IMT change was associated with apoB, total cholesterol, LDL cholesterol, triglycerides, and inversely with HDL cholesterol and LDL particle size at entry, and there was a strong colinearity between these variables. The subjects with apoB/apoA-I above the first tertile (0.74) had a 20-microm-higher (95% CI, 7 to 33) annual increase in IMT compared with those below this level after adjustment for blood pressure and smoking. CONCLUSIONS: The apoB/apoA-I ratio was strongly associated with MetS and its components at baseline. ApoB/apoA I at baseline was related to the change in carotid artery IMT during 3 years of follow-up. There was a strong colinearity between apoB/apoA and the atherogenic lipids. PMID- 15345796 TI - Findings from the reanalysis of the NINDS tissue plasminogen activator for acute ischemic stroke treatment trial. AB - BACKGROUND AND PURPOSE: Following publication of concerns about the results of the National Institute of Neurological Disorders and Stroke (NINDS) intravenous tissue plasminogen activator (t-PA) in acute stroke treatment trial, NINDS commissioned an independent committee "to address whether there is concern that eligible stroke patients may not benefit from t-PA given according to the protocol used in the trials and, whether the subgroup imbalance (in baseline stroke severity) invalidates the entire trial." METHODS: The original NINDS trial data were reanalyzed to assess the t-PA treatment effect, the effect of the baseline imbalance in stroke severity between the treatment groups on the t-PA treatment effect, and whether subgroups of patients did not benefit from receiving t-PA. RESULTS: A clinically important and statistically significant benefit of t-PA therapy was identified despite subgroup imbalances in baseline stroke severity and an increased incidence of symptomatic intracerebral hemorrhage in t-PA treated patients. The adjusted t-PA to placebo odds ratio (OR) of a favorable outcome was 2.1 (95% CI, 1.5 to 2.9). Although these exploratory analyses found no statistical evidence that the t-PA treatment effect differed among patient subgroups, the study was not powered to detect subgroup treatment differences. CONCLUSIONS: These findings support the use of t-PA to treat patients with acute ischemic stroke within 3 hours of onset under the NINDS t-PA trial protocol. Health professionals should work collaboratively to develop guidelines to ensure appropriate use of t-PA in acute ischemic stroke patients. PMID- 15345797 TI - The relative impact of inadequate primary and secondary prevention on cardiovascular mortality in the United States. AB - BACKGROUND AND PURPOSE: We developed a model to estimate the costs incurred by ineffective primary and secondary prevention in terms of excess cardiovascular disease (CVD) mortality in a nationally representative sample of the US population. METHODS: Cox proportional hazards analyses were used to examine the effect of inadequate risk factor control on the incidence of fatal stroke and myocardial infarction (MI) during a follow-up period of 13.4+/-3.6 years after adjusting for differences in age, gender, and ethnicity in a national cohort of 9252 adults who participated in the Second National Health and Nutrition Examination Survey (NHANES) Mortality Follow-up Study. Inadequate risk factor modification was defined by presence of either blood pressure >140/90 mm Hg, serum cholesterol >200 mg/dL, or active cigarette smoking. Using the data from 4115 adults screened in the NHANES 1999 to 2000, population attributable risk (PAR) percent and associated cost incurred (expressed as proportion of total 1 year cost incurred for CVD mortality in year 2001) was estimated. RESULTS: CVD mortality risk increased in a stepwise manner for persons with no previous MI or stroke and > or =2 inadequately controlled risk factors (2x); and previous history of MI and stroke and adequately controlled risk factors (2.6x), 1 inadequately controlled risk factor (4.3x), and > or =2 inadequately controlled risk factors (5.7x). The PAR was 14% (estimated cost incurred 13.2 billion dollars) among persons with > or =2 inadequately controlled risk factors without previous MI or stroke (estimated 17% of total US population). Among persons with previous MI or stroke, the PAR was 7% (cost incurred 6.2 billion dollars) and 8% (cost incurred 7.4 billion dollars) for 1 inadequately controlled risk factor and > or =2 inadequately controlled risk factors, respectively. An excess of cost of 13.6 billion dollars was spent on 4% of the total population (persons with inadequate secondary prevention). CONCLUSIONS: The model demonstrates the differential risk of mortality from inadequately controlled cardiovascular risk factors in primary and secondary prevention settings. The large financial cost incurred by inadequate primary and secondary prevention justifies intensive efforts directed toward detection and treatment of cardiovascular risk factors. PMID- 15345798 TI - Systematic review of computed tomographic angiography for assessment of carotid artery disease. AB - BACKGROUND AND PURPOSE: To review the literature on the diagnostic accuracy of computed tomographic angiography (CTA) compared with arteriography/intra-arterial digital subtraction angiography as reference standard for assessment of symptomatic carotid artery disease. METHODS: The PubMed, MEDLINE, PREMEDLINE, EMBASE, and CINAHL databases were searched to retrieve studies published between 1990 and July 2003, comparing CTA and intra-arterial digital subtraction angiography in patients with symptomatic carotid artery disease that presented raw data for detection of a <70% or 70% to 99% stenosis or an occlusion. Two observers independently assessed methodological quality and constructed 2x2 tables for sensitivity and specificity of CTA for detection of a 70% to 99% stenosis versus <70% stenosis or an occlusion, and for <99% stenosis versus occlusion. A bivariate random effects model was used to calculate the pooled sensitivity and specificity of CTA for detection of these lesions. RESULTS: Some 864 patients (66% male) with a mean age of 66 years were studied in the 28 studies included in the meta-analysis. In all studies, a single-slice CT-scan was used. Only 8 studies satisfied all methodological quality criteria. The pooled sensitivity and specificity for detection of a 70% to 99% stenosis were 85% (95% CI, 79% to 89%) and 93% (95% CI, 89% to 96%), respectively. For detection of an occlusion, the sensitivity and specificity were 97% (95% CI, 93% to 99%) and 99% (95% CI, 98% to 100%), respectively. Incomplete reporting of demographic characteristics and technical differences in the individual studies obstructed a meaningful subgroup analysis. CONCLUSIONS: CTA is an accurate modality for detection of severe carotid artery disease, especially for detection of occlusions. The fair methodological quality of the included studies must be taken into account when interpreting these results. PMID- 15345799 TI - Central nervous system entry of peripherally injected umbilical cord blood cells is not required for neuroprotection in stroke. AB - BACKGROUND AND PURPOSE: To date, stem cell graft-mediated neuroprotection is equated with graft survival and secretion of neurotrophic factors in the brain. Here, we examined whether neuroprotection by systemically delivered human umbilical cord blood (HUCB) cells was dependent on their entry into the central nervous system in a rodent model of acute stroke. METHODS: Adult male Sprague Dawley rats were subjected to right middle cerebral artery occlusion for 60 minutes. During the 1-hour occlusion, animals were randomly assigned to 1 of the following treatments: intravenous injection of HUCB (a subtherapeutic dose of 200,000 cells in 10 microL) with blood-brain barrier (BBB) permeabilizer (1.1 mol/L mannitol at 4 degrees C) or vehicle, intravenous vehicle alone, or intravenous mannitol alone. Behavioral tests, using elevated body swing test and passive avoidance test, were conducted at day 3 poststroke, and thereafter, animals were euthanized for: (1) immunohistochemical examination of HUCB, which were lentivirally labeled with green fluorescent protein; (2) cerebral infarction analysis using 2,3,5-triphenyl-tetrazolium chloride; and (3) enzyme-linked immunosorbent assay of trophic factors within the striatal region. RESULTS: We did not detect intravenously administered low dose of HUCB cells in the brains of animals at day 3 after stroke even when cells were coinfused with a BBB permeabilizer (mannitol). However, HUCB-mannitol treatment significantly increased brain levels of neurotrophic factors, which correlated positively with reduced cerebral infarcts and improved behavioral functions. CONCLUSIONS: Our data show that central nervous system availability of grafted cells is not a prerequisite for acute neuroprotection provided that therapeutic molecules secreted by these cells could cross the BBB. PMID- 15345800 TI - Fibrinogen concentration and risk of ischemic stroke and acute coronary events in 5113 patients with transient ischemic attack and minor ischemic stroke. AB - BACKGROUND AND PURPOSE: Fibrinogen is an independent risk factor for coronary events in population-based studies and in patients with coronary heart disease, but there is uncertainty about prediction of stroke, particularly in secondary prevention. METHODS: We studied unpublished data from 3 prospective studies of patients with recent transient ischemic attack (TIA) or minor ischemic stroke: the United Kingdom TIA Aspirin (UK-TIA) trial (n=1860); the Dutch TIA trial (n=2960); and the Oxford TIA Study (n=293). By separate and pooled analysis, we used Cox models to determine the relationship between fibrinogen and risk of ischemic stroke and other vascular events during 23,272 patient-years of follow up and adjusted for other risk factors. RESULTS: There was no significant heterogeneity in fibrinogen risk associations between studies. Fibrinogen predicted subsequent ischemic stroke, with a pooled hazard ratio (HR) for values above the median of 1.34 (95% CI, 1.13 to 1.60; P=0.001). The association tended to be stronger in patients with nonlacunar (HR=1.42; 95% CI, 1.13 to 1.78; P=0.002) than lacunar syndromes (HR=1.09; 95% CI, 0.80 to 1.49; P=0.58), but was not significantly so (P=0.18). There was no association with hemorrhagic stroke (adjusted HR=1.09; 95% CI, 0.55 to 2.17; P=0.81). Fibrinogen predicted acute coronary events (adjusted HR=1.42; 95% CI, 1.18 to 1.70; P<0.001) and all ischemic vascular events (adjusted HR=1.31; 95% CI, 1.15 to 1.49; P<0.001), but not nonvascular death (adjusted HR=1.24; 95% CI, 0.90 to 1.70; P=0.19). CONCLUSIONS: In patients with a previous TIA or ischemic stroke, risks of recurrent ischemic stroke and acute coronary events increase linearly with fibrinogen levels, but the relationships are weaker than in some previous population-based studies. PMID- 15345801 TI - Transplantation of circulating endothelial progenitor cells restores endothelial function of denuded rabbit carotid arteries. AB - BACKGROUND AND PURPOSE: Circulating endothelial progenitor cells (EPCs) play an important role in repair of injured vascular endothelium and neovascularization. The present study was designed to determine the effect of EPCs transplantation on the regeneration of endothelium and recovery of endothelial function in denuded carotid arteries. METHODS: Isolated mononuclear cells from rabbit peripheral blood were cultured in endothelial growth medium for 7 days, yielding EPCs. A rabbit model of common carotid artery denudation by passage of a deflated balloon catheter was used to evaluate the effects of EPCs on endothelial regeneration and vasomotor function. Immediately after denudation, autologous EPCs (10(5) cells in 200 microL saline) or 200 microL saline alone (control) were administered into the lumen of injured artery. RESULTS: Four weeks after transplantation, fluorescence-labeled colonies of EPCs were found in the vessel wall. Local transplantation of EPCs as compared with saline administration accelerated endothelialization and significantly improved endothelium-dependent relaxation when assessed 4 weeks after denudation (n=4 to 5, P<0.05). Transplantation of EPCs did not affect vasomotor function of arterial smooth muscle cells. Protein array analysis of conditioned media obtained from cultured EPCs demonstrated the ability of these cells to produce and release a number of proangiogenic cytokines. CONCLUSIONS: We conclude that local delivery of cultured circulating EPCs into the lumen of denuded carotid arteries accelerates endothelialization and improves endothelial function. Paracrine effects of EPCs may contribute to regenerative properties of EPCs. PMID- 15345802 TI - Complications and pitfalls in rat stroke models for middle cerebral artery occlusion: a comparison between the suture and the macrosphere model using magnetic resonance angiography. AB - BACKGROUND AND PURPOSE: Investigating focal cerebral ischemia requires animal models that are relevant to human stroke. Complications and side effects are common among these models. The present study describes potential pitfalls in 3 techniques for middle cerebral artery occlusion (MCAO) in rats using magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA). METHODS: Rats were subjected to temporary MCAO for 90 minutes using the suture technique (group I; n=10) or to permanent MCAO using the suture technique (group II; n=10) or the macrosphere technique (group III; n=10). Clinical evaluation was performed after 3 hours and 24 hours. After 24 hours, animals underwent MRI and MRA to determine lesion size and the intracranial vascular status. RESULTS: Hemispheric lesion volume was significantly smaller in group I (14.6%) compared with groups II (35.2%; P<0.01) and III (21.3%; P<0.05). Two animals (1 each in group II and III) did not demonstrate neurological deficits and had no lesion on MRI and a patent MCA main stem on MRA. Subarachnoid hemorrhage was detected in 2 animals (1 each in group I and II). MRA indicated a patent MCA main stem in 2 animals (group II), although both rats displayed neurological deficits. Hypothalamic infarction with subsequent pathological hyperthermia was detected in all animals in group II and in 1 rat in group III. CONCLUSIONS: Model failures occurred frequently in all groups. MRI and MRA helps to identify animals that need to be excluded from experimental stroke studies. PMID- 15345803 TI - Homocysteine and the risk of ischemic stroke in a triethnic cohort: the NOrthern MAnhattan Study. AB - BACKGROUND AND PURPOSE: The level of total homocysteine (tHcy) that confers a risk of ischemic stroke is unsettled, and no prospective cohort studies have included sufficient elderly minority subjects. We investigated the association between mild to moderate fasting tHcy level and the incidence of ischemic stroke, myocardial infarction, and vascular death in a multiethnic prospective study. METHODS: A population-based cohort was followed for vascular events (stroke, myocardial infarction, and vascular death). Baseline values of tHcy and methylmalonic acid were measured among 2939 subjects (mean age, 69+/-10; 61% women, 53% Hispanics, 24% blacks, and 20% whites). Cox proportional models were used to calculate hazard ratios (HRs) and 95% CIs in tHcy categories after adjusting for age, race, education, renal insufficiency, B12 deficiency, and other risk factors. RESULTS: The adjusted HR for a tHcy level > or =15 micromol/L compared with <10 micromol/L was greatest for vascular death (HR=6.04; 95% CI, 3.44 to 10.60), followed by combined vascular events (HR=2.27; 95% CI, 1.51 to 3.43), ischemic stroke (HR=2.01; 95% CI, 1.00 to 4.05), and nonvascular death (HR=2.02; 95% CI, 1.31 to 3.14). Mild to moderate elevations of tHcy of 10 to 15 micromol/L were not significantly predictive of ischemic stroke, but increased the risk of vascular death (2.27; 95% CI, 1.44 to 3.60) and combined vascular events (1.42; 95% CI, 1.06 to 1.88). The effect of tHcy was stronger among whites and Hispanics, but not a significant risk factor for blacks. CONCLUSIONS: Total Hcy elevations above 15 micromol/L are an independent risk factor for ischemic stroke, whereas mild elevations of tHcy of 10 to 15 micromol/L are less predictive. The vascular effects of tHcy are greatest among whites and Hispanics, and less among blacks. PMID- 15345805 TI - Cerebral hypoperfusion and amyloid-induced synergistic impairment of hippocampal CA1 synaptic efficacy and spatial memory in young adult rats. AB - Alzheimer's disease (AD) is characterized by amyloid plaques, neurofibrillary tangles in the brain, cerebral hypoperfusion/hypometabolism, and amyloid angiopathy. The former two and cell loss occur late in the disease and are probably not the leading causes for the initial memory decline. Cerebral hypoperfusion is a pre-clinical event in AD and represents the most accurate indicator predicting the probable AD patients to develop AD in a future time. However, in young animals, cerebral hypoperfusion as those matching the reduction in AD has no significant effects on learning and memory. Here, we report that association of cerebral hypoperfusion (2-vessel occlusion) with cerebrovascular amyloid (internal-carotid 0.5 mg beta(25-35), an active fragment of Abeta) significantly impaired spatial learning and memory of young adult rats, while neither the same insult alone had significant impact. At the time when the spatial memory was impaired, in vitro recording revealed that the associated cerebral hypoperfusion and internal-carotid amyloid reduced the ability of the hippocampal CA1 network to generate cholinergic theta and the synaptic modification evoked by associative activation of cholinergic and GABAergic inputs. The results suggest that cerebral hypoperfusion and amyloid angiopathy may play an important role as associated events in initiating the early memory decline in AD. PMID- 15345806 TI - Curcumin interaction with copper and iron suggests one possible mechanism of action in Alzheimer's disease animal models. AB - Curcumin is a polyphenolic diketone from turmeric. Because of its anti-oxidant and anti-inflammatory effects, it was tested in animal models of Alzheimer's disease, reducing levels of amyloid and oxidized proteins and preventing cognitive deficits. An alternative mechanism of these effects is metal chelation, which may reduce amyloid aggregation or oxidative neurotoxicity. Metals can induce Abeta aggregation and toxicity, and are concentrated in AD brain. Chelators desferrioxamine and clioquinol have exhibited anti-AD effects. Using spectrophotometry, we quantified curcumin affinity for copper, zinc, and iron ions. Zn2+ showed little binding, but each Cu2+ or Fe2+ ion appeared to bind at least two curcumin molecules. The interaction of curcumin with copper reached half-maximum at approximately 3-12 microM copper and exhibited positive cooperativity, with Kd1 approximately 10-60 microM and Kd2 approximately 1.3 microM (for binding of the first and second curcumin molecules, respectively). Curcumin-iron interaction reached half-maximum at approximately 2.5-5 microM iron and exhibited negative cooperativity, with Kd1 approximately 0.5-1.6 microM and Kd2 approximately 50-100 microM. Curcumin and its metabolites can attain these levels in vivo, suggesting physiological relevance. Since curcumin more readily binds the redox-active metals iron and copper than redox-inactive zinc, curcumin might exert a net protective effect against Abeta toxicity or might suppress inflammatory damage by preventing metal induction of NF-kappaB. PMID- 15345807 TI - Increase of white matter string vessels in Alzheimer's disease. AB - String vessels are collagenous structures connected to capillaries. They have no endothelial cells or lumen. We assessed collagen IV-labeled string vessels in the white matter (WM) of subjects with Alzheimer's disease (AD) (n = 12) and non-AD controls (n = 11) using 100 microm celloidin sections. Ten standard fields were digitally captured and the number and length of normal vessels and string vessels were quantified by computerized image analysis. The WM of the AD-diagnosed individuals contained more strings per mm2 (3.95 +/- 0.49) than comparable WM from controls (1.36 +/- 0.39) (p = 0.0005) and had increased total string vessel length in mm/mm2 (AD = 0.29 +/- 0.04; control = 0.10 +/- 0.03; p = 0.0015). There was a 25% increase (not statistically significant) in vessel density in mm/mm2 in AD subjects (AD = 11.88 +/- 0.87; control = 9.53 +/- 0.78; p = 0.06), presumably due to brain atrophy in the white matter. Although vessel length was slightly increased in AD subjects, they still had more than double the string length per total vessel length (AD = 2.88 +/- 0.38) compared to controls (1.36 +/- 0.27) (p = 0.0057). This increase in string vessels in the white matter of AD subjects suggests a decrease in vascular supply in this disease. PMID- 15345808 TI - Reduction of brain lipid peroxidation by CSF drainage in Alzheimer's disease patients. AB - Alzheimer's disease (AD) is associated with an increase in cerebrospinal fluid (CSF) levels of the isoprostane 8,12-iso-iPF2alpha-VI, a specific marker of in vivo lipid peroxidation. Poor cerebral clearance of end products of oxidative reactions via CSF circulation may contribute to and sustain ongoing stress. CSF drainage via a low-flow ventriculoperitoneal (VP) shunt may improve removal of these products, reducing oxidative stress. We quantified this biomarker in patients with AD undergoing to VP shunt placement at baseline and after one-year period. CSF sampling occurred at baseline and quarterly visits for one year. Levels of this isoprostane were determined simultaneously at the end of the study by gas chromatography negative ion chemical ionization mass spectrometry. Over one-year, CSF 8,12-iso-iPF2alpha-VI levels consistently decreased versus baseline (51% of initial level), while CSF protein, glucose, cell count and IgG concentrations remained within normal limits. This finding supports the hypothesis that improving CSF drainage enhances extra-cellular clearance of end products of oxidative reactions and lowers brain lipid peroxidation. PMID- 15345809 TI - Apolipoprotein E4 allele and ribosomal genes in Alzheimer's disease. AB - Ribosomal genes are involved in cellular transcription, translation and gene expression modulation process. An association between 28S/18S rRNA ratio levels with apoptosis and aging has been reported. Moulder et al. [22] and Hashimoto et al. [8] showed an association between apolipoprotein E4 allele and neuronal cell apoptosis through diverse mechanisms. The apoE 4 allele is considered a late onset Alzheimer's disease (AD) risk factor associated with AD pathogenesis. We evaluated the association between apoE4 allele genotyping by PCR and rRNA 28S/18S ratio by slot blotting technique using peripheral blood samples of 18 Alzheimer's disease patients, 18 elderly controls and 18 young controls. A rRNA ratio decrease was observed in AD individuals confirming our previous results but this association is independently of the ApoE4 allele genotype. Thus our results pointed that two different mechanisms are involved in the etiology of Alzheimer disease each one leading independently to cell death. Further studies could investigate these factors. PMID- 15345810 TI - Effects of the ApoE epsilon4 allele on olfactory function in Down syndrome. AB - The present study investigated the effects of the apolipoprotein E (ApoE) epsilon4 allele, a risk factor for Alzheimer's disease (AD), on olfactory function in Down syndrome (DS). Brain areas critical to olfactory processing, particularly the entorhinal cortex, show the earliest neuropathological changes in AD. Functionally, odor identification has been shown to be impaired in AD and in persons with the epsilon4 allele. DS is also a risk factor for AD. Thus, we hypothesized greater impairment in epsilon4 positive DS participants. Olfactory function was assessed with the San Diego Odor Identification Test in 34 participants with DS and 34 normal controls. Genomic DNA was prepared from blood samples to obtain ApoE status for the DS participants. Results indicate (1) that participants with DS had significant deficits in olfactory functioning; and (2) that among DS participants, those with an epsilon4 allele had poorer odor identification than those without an epsilon4 allele. The results support the hypothesis that individuals with DS who have an additional genetic risk factor for AD, the ApoE epsilon4 allele, exhibit greater deficits in odor identification. Areas of the brain involved in odor identification may be particularly affected in individuals with DS who carry the epsilon4 allele. PMID- 15345811 TI - Fruit extracts antagonize Abeta- or DA-induced deficits in Ca2+ flux in M1 transfected COS-7 cells. AB - Evidence suggests that there is a selective sensitivity to oxidative stress (OSS) among muscarinic receptor (MAChR) subtypes with M1, M2 and M4 showing > OSS than M3 or M5 subtypes in transfected COS-7 cells. This may be important in determining the regional specificity in neuronal aging and Alzheimer disease (AD). We assessed the effectiveness of blueberry (BB) and other high antioxidant (HA) fruit extracts (boysenberry, BY; cranberry, CB; black currant, BC; strawberry, SB; dried plums, DP; and grape, GR) on the toxic effects of Abeta 25 35 (100 microM, 24 hrs) and DA (1 mM, 4 hrs) on calcium buffering (Recovery) following oxotremorine (750 microM) -induced depolarization in M1AChR-transfected COS-7 cells, and on cell viability following DA (4 hrs) exposure. The extracts showed differential levels of Recovery protection in comparisons to the non supplemented controls that was dependent upon whether DA or Abeta was used as the pretreatment. Interestingly, assessments of DA-induced decrements in viability revealed that all of the extracts had some protective effects. These findings suggest that the putative toxic effects of Abeta or DA might be reduced by HA fruit extracts. PMID- 15345812 TI - Estrogen increases brain expression of the mRNA encoding transthyretin, an amyloid beta scavenger protein. AB - Estrogen replacement therapy in postmenopausal women is associated with a reduced risk of Alzheimer's Disease (AD). The multiple mechanisms by which estrogen protects against AD are still unknown. To conduct a broad screen for estrogen regulated AD-related genes in the brain, we used cDNA array assays of brain mRNA samples from ovariectomized (ovx) adult female mice treated with either 17beta estradiol or vehicle at 1 or 5 weeks post-ovx. The gene encoding transthyretin (TTR), which has been reported to scavenge amyloid beta peptides and reduce amyloid plaque formation, is increased by estradiol treatment at both 1 and 5 weeks post-ovx. Northern blot analyses and RNase protection assays performed on whole brain samples obtained from estradiol- or vehicle-treated mice confirmed the cDNA array assays showing a significant increase in TTR mRNA with estradiol treatment. Qualitative in situ hybridization or immunocytochemistry performed on brain sections demonstrated that TTR mRNA is expressed only in choroid plexus and leptomeninges, and that both estrogen receptor proteins, alpha and beta, are present in choroid plexus cells. These novel findings suggest that estrogen may reduce the risk of AD by acting on choroid plexus cells to increase TTR gene expression, leading to enhanced sequestration and reduced aggregation of amyloid beta peptides. PMID- 15345813 TI - Early changes in neurons of the hippocampus and neocortex in transgenic rats expressing intracellular human a-beta. AB - Alzheimer's disease (AD) studies typically focus on the extracellular impact of the amyloid-beta (Abeta) protein, however recent findings also implicate intracellular Abeta (iAbeta) accumulation in the disease's molecular neuropathology. In a double mutant transgenic rat model (AbetaPP and PS1 mutations, UKUR25), stably expressing intracellular human Abeta fragments in an environment devoid of both amyloid plaques and neurofibrillary tangles, we investigated the impact of iAbeta burden on both the incidence and relative cross sectional areas of the Golgi apparatus, lysosomes and lipofuscin bodies. Pyramidal cells within the hippocampus and neocortex of both transgenic and non transgenic age matched controls were compared. This comparison revealed a significant increase in both the proportional area occupied by Golgi apparatus elements as well as in the mean individual cross sectional area of Golgi compartments in the hippocampus of transgenic rats as compared to controls. Elevated lysosome and lipofuscin elements in the hippocampi of transgenic rats were observed, as was an increase in the mean individual, cross sectional area of lipofuscin bodies in the cortex of transgenic rats as compared to controls. These findings support the hypothesis that intracellular Abeta accumulation not only has an impact on subcellular compartments but also potentially contributes to the neuronal cell pathology observed in AD. PMID- 15345814 TI - Demonstration of a role for alpha-synuclein as a functional microtubule associated protein. AB - Alpha-synuclein is a major constituent of pathological intracellular inclusion bodies, a common feature of several neurodegenerative diseases. Two missense mutations in the alpha-synuclein gene have been identified in confirmed autosomal dominant familial Parkinson's disease, which segregate with the illness. However, the physiological function of alpha-synuclein remains unknown. After biochemical investigations we have revealed tubulin to be an alpha-synuclein associated/binding protein. Here, we show that alpha-synuclein induces polymerization of purified tubulin into microtubules. Mutant forms of alpha synuclein lose this potential. The binding site of alpha-synuclein to tubulin is identified, and co-localization of alpha-synuclein with microtubules is shown in cultured cells. To our knowledge, this is the first demonstration of microtubule polymerizing activity of alpha-synuclein. Now we can see a striking resemblance between alpha-synuclein and tau: both have the same physiological function and pathological features, making abnormal structures in diseased brains known as synucleinopathies and tauopathies. The discovery of a physiological role for alpha-synuclein may provide a new dimension in researches into the mechanisms of alpha-synuclein-associated neurodegenerative diseases. PMID- 15345815 TI - Alkali scandium arsenates. I. The framework structures of KSc(HAsO4)2 and RbScAs2O7. AB - The crystal structures of hydrothermally synthesized potassium scandium hydrogen arsenate(V), KSc(HAsO4)2, (I), and rubidium scandium diarsenate(V), RbScAs2O7, (II), were determined from single-crystal X-ray diffraction data collected at room temperature. Compound (I) represents a new microporous structure type, designated MCV-3, which is characterized by a three-dimensional framework of corner-sharing alternating ScO6 octahedra and HAsO4 tetrahedra. Intersecting tunnels parallel to [101] and [110] host eight-coordinate K atoms. There is one hydrogen bond of medium strength [O...O = 2.7153 (18) A]. Compound (II) is the first reported diarsenate with a KAlP2O7-type structure and is isotypic with at least 27 A(I)M(III) diphosphates. The average Sc-O bond lengths in (I) and (II) are 2.09 (2) and 2.09 (3) A, respectively. The K and Sc atoms in (I) lie on an inversion centre and a twofold axis, respectively. All atoms in (II) are in general positions. PMID- 15345816 TI - Alkali scandium arsenates. II. The framework structures of alpha- and beta CsSc(HAsO4)2. AB - The crystal structures of hydrothermally synthesized alpha-, (I), and beta caesium scandium bis[hydrogen arsenate(V)], (II), both CsSc(HAsO4)2, have been determined from single-crystal X-ray diffraction data collected at room temperature. The dimorphs are both characterized by a three-dimensional negatively charged framework of corner-sharing alternating ScO6 octahedra and HAsO4 tetrahedra. The charge-balancing Cs+ cations are located in a system of three intersecting tunnels in (I) and in tunnels parallel to the a axis in (II). Strong to weak hydrogen bonds reinforce both frameworks. The average Sc-O bond lengths are 2.098 and 2.094 A, respectively. Compound (I) is triclinic and isotypic with (NH4)Fe(III)(HPO4)2, alpha-A(I)V(III)(HPO4)2 (A is NH4 or Rb) and alpha-(NH4)(Al(0.64)Ga(0.36))(HPO4)2. Compound (II) is monoclinic and isotypic with (H3O)Fe(III)(HPO4)2, beta-A(I)V(III)(HPO4)2 (A is NH4 or Rb), CsIn(HPO4)2 and RbSc(HPO4)2. Both (I) and (II) represent the first arsenate examples of their structure types. The Cs and Sc atoms in (I) lie on inversion centres. In (II), all atoms are in general positions. A brief overview is presented of the six structure types shown by A(I)M(III)(HXO4)2 compounds (X is P or As). PMID- 15345817 TI - A new chromate of tetravalent cerium: Ce2(CrO4)4.2H2O. AB - Dicerium(IV) tetrachromate(VI) dihydrate, Ce2(IV)(CrO4)4.2H2O, has been prepared from an acidic aqueous solution at room temperature. Its novel crystal structure, which was solved from single-crystal X-ray diffraction data, is built from isolated CrO4 tetrahedra and isolated Ce(O,H2O)n (n = 8 and 9) polyhedra. All atoms are in general positions. The mean Ce-O and Cr-O bond lengths are 2.358 and 1.651 A, respectively. Comparisons are drawn with the structure of Ce(IV)(CrO4)2.2H2O. PMID- 15345818 TI - Powder neutron diffraction of SrNbO2N at room temperature and 1.5 K. AB - The structure of strontium niobium dioxygen nitride, SrNbO2N, has been solved by powder neutron diffraction at room temperature and 1.5 K. SrNbO2N crystallizes in the tetragonal space group I4/mcm, with a = 5.7056 (4) and c = 8.1002 (9) A at room temperature, and a = 5.6938 (4) and c = 8.0974 (8) A at 1.5 K. The crystal structure is derived from the cubic perovskite archetype by a slight rotation of the Nb(O,N)6 octahedra with respect to the tetragonal axis. A partially ordered distribution of oxygen and nitrogen on the anionic sites was found. PMID- 15345819 TI - Mg7(AsO4)2(HAsO4)4: a new magnesium arsenate with a very strong hydrogen bond. AB - A new compound, heptamagnesium bis(arsenate) tetrakis(hydrogenarsenate), Mg7(AsO4)2(HAsO4)4, was synthesized by a hydrothermal method. The structure is based on a three-dimensional framework of edge- and corner-sharing MgO6, MgO4(OH)2, MgO5, AsO3(OH) and AsO4 polyhedra. Average Mg-O and As-O bond lengths are in the ranges 2.056-2.154 and 1.680-1.688 A, respectively. Each of the two non-equivalent OH groups is bonded to both an Mg and an As atom. One OH group is involved in a very short hydrogen bond [O.O = 2.468 (3) A]. The formula unit is centrosymmetric, with all atoms in general positions except for one Mg atom, which has site symmetry -1. The compound is isotypic with Mn7(AsO4)2(HAsO4)4 and M7(PO4)2(HPO4)4, where M is Fe, Co or Mn. PMID- 15345820 TI - Bis[1,3-bis(2-methyltetrazol-5-yl-kappaN4)triazenido-kappaN2]nickel(II). AB - The title compound, [Ni(BMTT)2], where BMTT is 1,3-bis(2-methyltetrazol-5 yl)triazenide (C4H6N11), presents a molecular complex with tridentate ligands. The tridentate mode of the ligand is realised through the central N atom of the triazene group and two N atoms of the two tetrazole rings. The [Ni(BMTT)2] molecule is the meridional isomer, with crystallographic 4 symmetry in space group P4(2)/n. The nickel centre has a distorted octahedral environment, with two axial Ni-N bonds of 2.041 (2) A and four equatorial Ni-N bonds of 2.0739 (14) A. The molecules are linked together by van der Waals interactions only. PMID- 15345821 TI - Bis(mu-2-sulfonatobenzoato)bis[(1,10-phenanthroline)lead(II)] dihydrate. AB - In the title centrosymmetric dimer, [Pb2(sbc)2(phen)2].2H2O [sbc is the 2 sulfonatobenzoate dianion (C7H4O5S) and phen is 1,10-phenanthroline (C12H8N2)], each Pb(II) ion is six-coordinated by four O atoms, viz. carboxylate and sulfonate O atoms from two sbc anions, and two N atoms from a 1,10-phenanthroline ligand. One 1,10-phenanthroline ligand and the carboxylate group of one sbc ligand are chelated to each Pb(II) cation, and the sulfonate group of the other sbc unit is monodentate. One O atom of the chelated carboxylate group also bridges to the other Pb(II) cation, so that each pair of Pb(II) ions is bridged by two sbc anions and has the same coordination environment, forming a dinuclear ring. Each pair of Pb(II) ions is thus connected by two different kinds of bridges, namely a carboxylate short bridge and a carboxylate-sulfonate long bridge. There is also a special position of 1 site symmetry at the centre of the two Pb(II) cations. PMID- 15345822 TI - Two phases of bis(tetraethylammonium) di-mu-chloro-bis[dichloropalladium(II)]. AB - A phase transition was found to occur at approximately 153 K in the title compound, (C8H20N)2[PdCl6]. The structures of the two phases are reported at 292 and 130 K. The low-temperature phase is twinned. The phase transition is accompanied by a minor displacement of the ions. There are C-H...Cl interactions as short as approximately 2.80 A, indicating the existence of hydrogen bonds, and this was confirmed by vibrational spectroscopy. The [Pd2Cl6]2- anion occupies sites of mmm and 2/m symmetry in the room-temperature and low-temperature phases, respectively. PMID- 15345823 TI - An unexpected chelate conformation in trans-[(R)-N,N'-bis(alpha phenylsalicylidene)propane-1,2-diaminato(2-)]bis(1-methyl-1H-imidazole kappaN3)cobalt(III) perchlorate. AB - The title compound, [Co(C29H24N2O2)(C4H6N2)2]ClO4, contains an optically active tetradentate Schiff base ligand in an equatorial plane and two 1-methylimidazole ligands at apical positions. The central N-C-C-N chelate ring of the Schiff base ligand has an envelope structure with a lambda conformation, which is different from the solution structure predicted from circular dichroism and 1H NMR spectra. PMID- 15345824 TI - Bis(4,4'-bipyridine-kappaN)bis(hydrogen phthalato)-kappa2O,O';kappaO-zinc(II). AB - The title compound, [Zn(C8H5O4)2(C10H8N2)2], was obtained by the hydrothermal reaction of ZnSO4.7H2O with phthalic acid (H2pht) and 4,4'-bipyridine (4,4' bipy). Crystallographic analysis shows that it has a one-dimensional double-chain structure via hydrogen-bonding interactions. Each Zn(II) atom, adopting a distorted tetrahedral geometry, is coordinated by two N atoms from two 4,4'-bipy ligands, with Zn-N distances of 2.054 (4) and 2.104 (4) A, and by two O atoms from symmetry-related Hpht- ligands, with Zn-O distances of 1.921 (4) and 2.019 (4) A. PMID- 15345825 TI - Bis(mu-cystamine-kappa4N,S:S',N')bis[(2-aminoethanethiolato-kappa2N,S) iridium(III)] tetrabromide dihydrate. AB - In the complex cation of the title compound, [Ir2(C2H6NS)2(C4H12N2S2)2]Br4.2H2O, which was obtained by rearrangement of [Re[Ir(aet)3]2]3+ (aet is 2 aminoethanethiolate) in an aqueous solution, two approximately octahedral fac(S) [Ir(NH2CH2CH2S)3] units are linked by two coordinated disulfide bonds. The complex cation has a twofold axis, and the two non-bridging thiolate S atoms in the complex are located on opposite sides of the two disulfide bonds. Considering the absolute configurations of the two octahedral units (Delta and Lambda) and the four asymmetric disulfide S atoms (R and S), the complex consists of the Delta(RR)Delta(RR) and Lambda(SS)Lambda(SS) isomers, which combine to form the racemic compound. PMID- 15345826 TI - catena-Poly[[[mu-pyridinium-3-carboxylato-bis[dioxomolybdenum(VI)]]-di-mu3-oxo] monohydrate]. AB - A novel chain molybdenum compound, [[Mo2O6(C6H5NO2)].H2O]n, which was synthesized under hydrothermal conditions, consists of an infinite rail-like chain formed by molybdenum oxide units linked by zwitterionic nicotinic acid ligands. Each Mo atom is coordinated octahedrally by six O atoms and the MoO6 octahedra are linked to one another via edge-sharing to produce a zigzag polymeric chain, with nicotinic acid ligands located, alternately, on each side of the rail-like chain plane. PMID- 15345827 TI - Mu-biphenyl-2,2'-dithiolato-kappa2S:S'-bis[(triphenylphosphine-kappaP)gold(I)]. AB - The reaction of ClAuPPh3 and 1,1'-biphenyl-2,2'-dithiol in the presence of trimethylbenzylammonium chloride and K2CO3 in a tetrahydrofuran/methanol solution gives the title complex, [Au2(C12H8S2)(C18H15P)2]. The molecule contains P-Au-S units which 'cross' with torsion angles of approximately 90 degrees [P-Au-Au-P = 86.23 (5) degrees and S-Au-Au-S = 95.62 (5) degrees]. The intra- and intermolecular Au.Au distances [3.9064 (3) and 6.3797 (5) A, respectively] are outside the range for typical Au...Au interactions. However, the Au atoms appear to be drawn together, leading to a significant bending of the P-Au-S angles [170.24 (5) and 169.52 (5) degrees]. PMID- 15345828 TI - A one-dimensional ladder-like coordination polymer derived from chains formed via hydrogen bonds: catena-poly[[aquadipyridinenickel(II)]-mu-2,2'-dithiodibenzoato kappa3O,O':O"]. AB - The title one-dimensional chain nickel(II)-disulfide complex, [Ni(C14H8O4S2)(C5H5N)2(H2O)]n, has each Ni(II) cation coordinated by two N atoms from two pyridine ligands, three carboxylate O atoms from two different dithiodibenzoate ligands and one O atom from a coordinated water molecule, in a distorted octahedral coordination geometry. Each dithiodibenzoate ion links two Ni(II) cations through its carboxylate O atoms, making the structure polymeric. Hydrogen-bond interactions between two shoulder-to-shoulder chains lead to the formation of a ladder-like structure. PMID- 15345829 TI - Bis[mu-2-[3 (cyclohexylammonio)propyliminomethyl]phenolato]bis[dichlorocopper(II)]. AB - The title compound, [Cu2(C16H24N2O)2Cl4], is a dinuclear copper(II) complex with inversion symmetry. Each Cu(II) atom is five-coordinated by two O and one N atom from two Schiff base ligands, and by two Cl atoms, giving an approximately trigonal-bipyramidal coordination environment. PMID- 15345830 TI - catena-Poly[1,4-diazoniabicyclo[2.2.2]octane [silver(I)-tri-mu-thiocyanato kappa6S:S]]. AB - The title compound, [(C6H14N2)[Ag(NCS)3]]n, is a polymeric silver(I) complex. The Ag(I) atom is hexacoordinated by the S atoms of six thiocyanate anions, with each thiocyanate S atom acting in a bridging mode to link the Ag atoms together. The unique Ag(I) atom lies at a cell origin and has crystallographically imposed 3 symmetry. The diazonia[2.2.2]octane molecule lies about a site with 6 imposed symmetry with the unique N atom on a threefold axis. The S and N atoms of the thiocyanate ligands sit on a mirror plane and a threefold axis, respectively. The crystal structure consists of one-dimensional chains, which are stabilized by N H...N hydrogen bonds to form a three-dimensional network. PMID- 15345831 TI - [Pd(CH3COO)2]n from X-ray powder diffraction data. AB - The water-insoluble title compound, catena-poly[palladium(II)-di-mu-acetato kappa4O:O'], [Pd(C2H3O2)2]n, was obtained from a nitratopalladium solution and acetic acid as a pale-pink powder. Ab initio crystal structure determination was carried out using X-ray powder diffraction techniques. Patterson and Fourier syntheses were used for atom location and the Rietveld technique was applied for the final structure refinement. The structure consists of palladium acetate complexes connected into polymeric chains running along b, in which two Pd atoms are bridged by two acetate groups that are in a cis configuration with respect to one another. The unique Pd atom lies on a site with 2/m symmetry and the acetate moieties have imposed m symmetry; these are joined into infinite chains running along the b direction. The shortest Pd...Pd distance in the row is 2.9192 (1) A. The planes of adjacent palladium complexes are inclined towards each other, the angle between the planes being approximately 30 degrees. PMID- 15345832 TI - Tetraaqua(3,4,7,8-tetramethyl-1,10-phenanthroline-kappa2N,N')zinc(II) thiosulfate. AB - In the title complex of zinc(II) with 3,4,7,8-tetramethyl-1,10-phenanthroline (tmph), viz. [Zn(C16H16N2)(H2O)4](S2O3), the metal atom has a monomeric octahedral ZnN2O4 complex environment comprising two N-atom donors from the tmph group and four aqua O-atom donors. The complex cation is connected to four thiosulfate anions through a compact hydrogen-bonding network involving all coordinated aqua H-atom donors and all the outer acceptors (O and S) of the anion. PMID- 15345833 TI - Aquabis(2-nitrobenzoato-kappaO)(1,10-phenanthroline-kappa2N,N')zinc(II). AB - The title compound, [Zn(C7H4NO4)2(C12H8N2)(H2O)], has been synthesized. X-Ray analysis reveals that it is a neutral zinc(II) mononuclear carboxylate complex based on mixed N- and O-donor ligands. The Zn atom is five-coordinate in a distorted trigonal-bipyramidal coordination environment involving two O atoms of two monodentate 2-nitrobenzoate molecules, two N atoms of a 1,10-phenanthroline molecule and one O atom of a water molecule. The axial positions are occupied by a carboxylate O atom from the 2-nitrobenzoate ligand and an N atom from the 1,10 phenanthroline ligand [N-Zn-O = 167.66 (9) degrees]. PMID- 15345834 TI - Bis(micro-ethyl pyridine-2,6-dicarboxylato)bis[diaquachlorocadmium(II)] dihydrate. AB - The reaction of cadmium chloride with pyridine-2,6-dicarboxylic acid (PDA) and 98% H2SO4 in ethanol led to the formation of the title compound, bis[mu-6 (ethoxycarbonyl)pyridine-2-carboxylato]-1:2kappa4O6,N,O2:O2;1:2kappa4O2:O2,N,O6 bis[diaquachlorocadmium(II)] dihydrate, [Cd2(C9H8NO4)2Cl2(H2O)4].2H2O. PDA is esterified to monoethyl pyridine-2,6-dicarboxylate (MEPD) by the catalysis of H2SO4 during the reaction. The dinuclear Cd(II) complex lies about an inversion centre and the unique Cd atom has a pentagonal-bipyramidal geometry. The two Cd atoms are bridged by two carboxylate O atoms, forming a planar Cd2O2 unit. Stair like chains are formed via O-H.Cl hydrogen bonds and these are further arranged into two-dimensional layers via hydrogen bonds involving solvate water molecules. PMID- 15345835 TI - Poly[[bis[aqua(1,10-phenanthroline)lanthanum(III)]-mu-aqua-di-mu-5 sulfonatoisophthalato] monohydrate] and poly[[[triaqualanthanum(III)]-mu-5 sulfonatoisophthalato] monohydrate]. AB - In the two related polymeric title compounds, [[La2(sip)2(phen)2(H2O)3].H2O]n [sip is the 5-sulfonatoisophthalate trianion (C8H3O7S3-) and phen is 1,10 phenanthroline (C12H8N2)], (I), and [[La(sip)(H2O)3].H2O]n, (II), the lanthanum(III) ions are nine-coordinate, with similar distorted monocapped square antiprism coordination geometry. The two crystal structures are very different. In (I), the sip anion acts as a pentadentate ligand, one of the coordinated water molecules lies on a twofold axis and further inversion, n-glide and translation operations generate a two-dimensional framework. In (II), the sip anion functions as a hexadentate ligand and a three-dimensional network with trinuclear 24 membered rings is developed via inversion, n-glide, twofold-screw and translation operations. Both structures also have extensive O-H...O hydrogen-bonded networks and pi-pi interactions. PMID- 15345836 TI - Amidobis[eta5-1,3-bis(trimethylsilyl)cyclopentadienyl]titanium(III). AB - The title compound, [Ti(C11H21Si2)2(NH2)], crystallizes as a bent metallocene, with the NH2 group oriented to maximize the pi interaction between the N atom and the Ti(III) center. The increased pi bonding is reflected in a short Ti-N bond length of 1.933 (3) A. PMID- 15345837 TI - Three 5H-indeno[1,2-c]pyridazin-5-one derivatives, potent type-B monoamine oxidase inhibitors. AB - The structures of three compounds, namely 7-methoxy-2-[3-(trifluoromethyl)phenyl] 9H-indeno[1,2-c]pyridazin-9-one, C19H11F3N2O2, (Id), 6-methoxy-2-[3 (trifluoromethyl)phenyl]-9H-indeno[1,2-c]pyridazin-9-one, C19H11F3N2O2, (IId), and 2-methyl-6-(4,4,4-trifluorobutoxy)-9H-indeno[1,2-c]pyridazin-9-one, C16H13F3N2O2, (IIf), which are potent reversible type-B monoamine oxidase (MAO-B) inhibitors, are presented and discussed. Compounds (Id) and (IId) crystallize in a nearly planar conformation. The crystal structures are stabilized by weak C H...O hydrogen bonds. The packing is dominated by pi-pi stacking interactions between the heterocyclic central moieties of centrosymmetrically related molecules. In compound (IIf), the trifluoroethyl termination is almost perpendicular to the plane of the ring. PMID- 15345838 TI - N-(3H-thiazol-2-ylidene)nitramine and N-methyl-N-(thiazol-2-yl)nitramine. AB - The geometries of the thiazole ring and the nitramino groups in N-(3H-thiazol-2 ylidene)nitramine, C3H3N3O2S, (I), and N-methyl-N-(thiazol-2-yl)nitramine, C4H5N3O2S, (II), are very similar. The nitramine group in (II) is planar and twisted along the C-N bond with respect to the thiazole ring. In both structures, the asymmetric unit includes two practically equal molecules. In (I), the molecules are arranged in layers connected to each other by N-H...N and much weaker C-H...O hydrogen bonds. In the crystal structure of (II), the molecules are arranged in layers bound to each other by both weak C-H...O hydrogen bonds and S...O dipolar interactions. PMID- 15345839 TI - 6beta-Azido-7alpha-hydroxy-17-oxo-5alpha-androstan-3beta-yl acetate. AB - In the title compound, C21H31N3O4, a potential inhibitor of aromatase, all rings are fused trans. Rings A, B and C have chair conformations which are slightly flattened. Ring D has a 14alpha-envelope conformation. The steroid nucleus has a small twist, as shown by the C19-C10...C13-C18 torsion angle of 6.6 (2) degrees. Ab initio calculations of the equilibrium geometry of the molecule reproduce this small twist, which appears to be due to the steric effect of the 6beta-azide substituent rather than to packing effects. PMID- 15345840 TI - The 1:1 adduct of hexamethylenetetramine (HMT) with racemic trans-1,2 cyclohexanedicarboxylic acid (CDA). AB - Hexamethylenetetramine and rac-trans-1,2-cyclohexanedicarboxylic acid crystallize in a 1:1 ratio as a neutral molecular adduct, C6H12N4.C8H12O4. Two dicarboxylic acid molecules and two tetramine molecules form a hydrogen-bonded ring, in the shape of a rhombus, which lies on a crystallographic twofold axis bisecting the two diacid molecules. The O-H...N hydrogen bonds have lengths 2.6808 (19) and 2.6518 (19) A, and, in each ring, both acid molecules have the same handedness. PMID- 15345841 TI - Two biologically active thiophene-3-carboxamide derivatives. AB - The compounds 2-[[(E)-(4-methoxyphenyl)methylene]amino]-N-(3-methylphenyl) 4,5,6,7-tetrahydro-1-benzothiophene-3-carboxamide, C24H24N2O2S, (I), and N-(4 methylphenyl)-2-[[(E)-(4-methylphenyl)methylene]amino]-4,5,6,7-tetrahydro-1 benzothiophene-3-carboxamide, C24H24N2OS, (II), show antibacterial and antifungal activities. The m-toluidine ring in (I) and the p-toluidine ring in (II) are coplanar with their respective thiophene rings. In (I), an intermolecular C-H...O hydrogen bond is present, whereas (II) does not exhibit any significant intermolecular interactions. However, in both compounds, an intramolecular N-H.N hydrogen bond forms a pseudo-six-membered ring, thus locking the molecular conformation and eliminating conformational flexibility. PMID- 15345842 TI - 4',4',6',6'-Tetrachloro-3,4-dihydro-3-(6-methylpyridin-2-yl)spiro[1,3,2 benzoxazaphosphinine-2,2'-(2lambda5,4lambda5,6lambda5-cyclotriphosphazene)]. AB - The title compound, C13H12Cl4N5OP3, is a phosphazene derivative with a bulky substituted spirocyclic ring. The C3NPO spirocyclic ring has a twist-boat conformation, while the phosphazene ring has a very flattened boat conformation. PMID- 15345843 TI - Diethyl 3,8-dimethyl-4,7-diazadeca-2,8-dienedioate. AB - The title compound, C14H24N2O4, consists of two symmetric moieties related through a twofold axis. The whole molecule has a cis conformation. Both the ionic enol form and the non-ionic keto form make comparable contributions to the structure. In the crystal structure, infinite supramolecular chains are formed through N-H...O hydrogen bonds. PMID- 15345844 TI - Crystal engineering via C-H...F and C-H...pi interactions in two substituted indoles. AB - The crystal structures of 1-(4-fluorophenyl)-2-phenyl-4,5,6,7-tetrahydro-1H indole, C20H18FN, and 1-(4-fluorophenyl)-6,6-dimethyl-2-phenyl-4,5,6,7-tetrahydro 1H-indole, C22H22FN, have been determined in order to study the role of 'organic fluorine' in crystal engineering. These molecules pack in the crystal structure via different types of molecular motifs utilizing weak C-H...F and C-H...pi interactions. PMID- 15345845 TI - 1-[4-(Methylsulfonyl)phenyl]-5-phenyl-1H-pyrazole derivatives. AB - Three related compounds containing a pyrazole moiety with vicinal phenyl rings featuring a methylsulfonyl substituent are described, namely 3-methyl-1-[4 (methylsulfonyl)phenyl]-5-phenyl-1H-pyrazole, C17H16N2O2S, ethyl 1-[4 (methylsulfonyl)phenyl]-5-phenyl-1H-pyrazole-3-carboxylate, C19H18N2O4S, and 1-[4 (methylsulfonyl)phenyl]-3-[3-(morpholino)phenoxymethyl]-5-phenyl-1H-pyrazole, C27H27N3O4S. The design of these compounds was based on celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, in order to study the influence of various substituents on COX-2 and 5-lipoxygenase (5-LOX) inhibition. PMID- 15345846 TI - Secondary interactions in the isomorphous compounds 2,6 bis(chloromethyl)pyridinium chloride and 2,6-bis(bromomethyl)pyridinium bromide. AB - The title compounds, C7H8Cl2N+.Cl- and C7H8Br2N+.Br-, are isomorphous. In the crystal packing, layers parallel to the ac plane are formed by a classical N(+) H...X- hydrogen bond (X = halogen) and two X...X contacts. A third X...X contact links the layers, and a fourth, which is however very long, completes a ladder like motif of halogen atoms. Hydrogen bonds of the form C-H...X play at best a subordinate role in the packing. PMID- 15345847 TI - Preference for the Diels-Alder addition of dienes syn to the O atom in cross conjugated spirocyclic cyclohexadienones. AB - In the Diels-Alder reaction, the preferred addition of dienes syn to the O atom in cross-conjugated cyclohexadienones containing an oxa-spiro ring system is observed. The two structures reported here, namely rel-(1R,4aR,9S,9aS,10R) 4a,9,9a,10-tetrahydro-9,10-diphenylspiro[9,10-epoxyanthracene-1(4H),2'-oxiran]-4 one, C27H20O3, and rel-(1R,4aS,9R,9aS,10S)-4a,9,9a,10-tetrahydro-9,10 diphenylspiro[9,10-epoxyanthracene-1(4H),2'-oxetane]-4-one, C28H22O3, are the minor and sole products, respectively, of the reactions of diphenylisobenzofuran with two slightly different cyclohexadienones. These structures differ in the size of the oxa-spiro ring, by one C atom, and in the relative configuration at the spirocyclic ring C atom, leading to some minor conformational differences between the two compounds. PMID- 15345848 TI - (Z)-2-(1-Phenylsulfonyl-1H-indol-3-ylmethylene)-1-azabicyclo[2.2.2]octan-3-one and (Z)-(S)-2-(1-phenylsulfonyl-1H-indol-3-ylmethylene)-1-azabicyclo[2.2.2]octan 3-ol. AB - The title compounds, C22H20N2O3S, (I), and C22H22N2O3S, (II), crystallize in space groups P-1 and P2(1)2(1)2(1), respectively. The indole rings are planar and the benzene ring of the phenylsulfonyl group makes a dihedral angle with the mean plane of the indole ring of 90.2 (2) degrees in (I) and 94.0 (2) degrees in (II). In both molecules, the double bond connecting the aza-bicyclic and indole moieties has a Z geometry. Compound (II) was obtained as an enantiomerically pure crystal and has the 3S configuration. PMID- 15345849 TI - 6-(4-Methoxybenzylamino)purin-3-ium chloride. AB - The title compound, C13H14N5O+.Cl-, belongs to the group of aromatic cytokinins. These compounds affect a variety of important physiological processes in plants and animals as well as in bacteria, including cell division, differentiation and senescence. The structure consists of a 6-(4-methoxybenzylamino)purinium cation and a Cl- anion. The cation moiety exists as the N3-protonated N7 tautomer. The cation contains nearly planar benzene and purine ring systems, with a dihedral angle of 77.46 (5) degrees. The crystal structure is stabilized by N(amino) H...N(purine) hydrogen bonds connecting two adjacent molecules, thus forming centrosymmetric dimers. PMID- 15345850 TI - 3,6-Bis(2-pyridyl)di-1,2,4-triazolo[3,4-a:4',3'-c]phthalazine. AB - The title compound, C20H12N8, (I), has been prepared by the reaction of 1,4 dihydrazinophthalazine and pyridine-2-carboxaldehyde, followed by an oxidative cyclization by treatment with bromine. In the solid state, the molecules of (I) are discrete, comprising a fused and twisted four-ring system with an overall helical appearance. The distance between the two intramolecular pyridyl N atoms is 3.075 (2) A, this short contact distance suggesting a pi-pi interaction. PMID- 15345851 TI - Desorption kinetics of a xanthenol-dioxane clathrate. AB - The host xanthenol compound forms a 1:1 clathrate with dioxane, namely 9-(1 naphthyl)-9H-xanthen-9-ol-1,4-dioxane, C23H16O2.C4H8O2. The structure of this clathrate is reported, along with a study of the kinetics of desolvation and the determination of an activation energy. The guest molecules are stabilized by O(host)-H...O(guest) hydrogen bonds [O-H = 0.968 (2) A, O...O = 2.7532 (13) A and O-H...O = 151.9 (4) degrees]. PMID- 15345852 TI - Structure-activity relationships in 4- and 5-androstene: 3beta-acetoxy-17-methyl 17-oxo-16,17-seco-5-androstene-16-carbonitrile and 17-methyl-3,17-dioxo-16,17 seco-4-androstene-16-carbonitrile. AB - The title compounds, C22H31NO3 and C20H27NO2, have similar conformations except in the molecular geometry and the bonding of two of the rings. These differences lead to marked differences in the biological activities of these compounds. Molecules of both compounds are linked by weak C-H...O hydrogen bonds in the crystal structures. PMID- 15345853 TI - 1-Methyl-1-phenyl-3-[1-hydroxyimino-2-(succinimido)ethyl]cyclobutane. AB - In the title compound, C17H20N2O3, the cyclobutane ring is puckered, with a dihedral angle of 19.11 (15) degrees. The 1-phenyl and 3-[1-hydroxyimino-2 (succinimido)ethyl] groups are in cis positions. The molecules are linked by O H...O and C-H...pi(benzene) interactions, forming a two-dimensional network. PMID- 15345854 TI - 2-Amino-2-thiazoline and its 1:1 organic salt with 2-naphthoxyacetic acid. AB - The crystal structures of 2-amino-2-thiazoline, C3H6N2S, and 2-amino-2 thiazolinium 2-naphthoxyacetate, C3H7N2S+.C12H9O3-, are reported. The structure of 2-amino-2-thiazoline consists of two unique molecules that construct a convoluted hydrogen-bonded ribbon involving R(2)2(8) graph-set association via both N-H...N and N-H...S interactions. The organic salt structure consists of the two molecules associated via an R(2)2(8) graph-set dimer through N-H...O interactions, with the hydrogen-bonding network propagated via additional N-H...O three-centre interactions from the second 2-amine H atom. PMID- 15345855 TI - Dimethyl 2-(4-bromophenyl)-10,11-dimethoxy-2,3,7,8-tetrahydrospiro[azepino[2,1 a]isoquinoline-3,9'-fluorene]-4,5-dicarboxylate. AB - The title compound, C38H32BrNO6, is a new photochromic tetrahydroazepinoisoquinoline (THAI). The longest spiro bond [1.589 (4) A] can be broken very easily by UV light, leading to ring opening. This explains the photochromic behaviour. PMID- 15345856 TI - Orthorhombic and monoclinic polymorphs of 1,3,5-triphenylperhydro-1,3,5-triazine 2,4,6-trione at 120 K: chains and sheets formed by C-H...pi(arene) hydrogen bonds. AB - The title compound, C21H15N3O3, crystallizes in two polymorphic forms. In the orthorhombic polymorph, (I), in space group Fdd2 with Z' = 1, the molecules lie in general positions, while in the monoclinic polymorph, (II), in space group C2/c with Z' = 1/2, the molecules lie across twofold rotation axes. In each polymorph, the molecules are linked by a single C-H.pi(arene) hydrogen bond, forming chains in polymorph (I) and sheets in (II). PMID- 15345857 TI - Hydrogen-bonded dimers in 2-nitrobenzaldehyde hydrazone and a three-dimensional hydrogen-bonded framework in 3-nitrobenzaldehyde hydrazone. AB - Molecules of 2-nitrobenzaldehyde hydrazone, C7H7N3O2, where Z' = 2, are linked by two N-H...N hydrogen bonds into isolated dimers, whereas in the isomeric 3 nitrobenzaldehyde hydrazone, where Z' = 1, the molecules are linked by one N H...O hydrogen bond and one N-H...N hydrogen bond into a three-dimensional framework structure. PMID- 15345858 TI - 1-Methylpiperazine-1,4-diium 4-nitrophthalate(2-) 4-nitrophthalic acid monohydrate. AB - The title adduct, C5H14N2(2+).C8H3NO6(2-).C8H5NO6.H2O, crystallizes in the monoclinic space group P2(1). All O atoms of the 4-nitrophthalate anions and neutral 4-nitrophthalic acid molecules are involved in hydrogen bonding with the piperazine dication and the water molecule of crystallization. PMID- 15345859 TI - C-H...pi, pi-pi and C-H...Cl interactions in chloro-substituted Schiff bases and 4-chloro-N-[4-(dimethylamino)benzylidene]aniline. AB - Molecular packing analyses were carried out on 15 crystal data sets of chloro substituted Schiff bases, including that of the title compound, C15H15ClN2. C H...pi and pi-pi interactions play a major role in the molecular self-assembly in the crystal. The former interactions favor molecules assembling into a screw, with a non-centrosymmetric crystal structure. When the molecular dipole is small, pi-pi interactions favor a parallel, but not usually antiparallel, mode of packing. Weak C-H...X hydrogen bonds (X = Cl or Br) and X...X interactions seem to be a secondary driving force in packing. The title molecule takes the trans form and the two benzene rings are twisted around the central linkage in opposite directions. In the crystal structure, molecules interact through C-H...pi and pi pi interactions, forming a 'dimer' and further forming double chains along [001]. The double chains are extended along [101] through C-H...Cl hydrogen bonds, forming double layers in (010). In the third direction, there are only ordinary, weaker, van der Waals interactions, which explains the crystal habit (i.e. thin plate). PMID- 15345860 TI - 1-(4-Bromophenyl)-3-(4-methylphenyl)prop-2-en-1-one. AB - In the molecule of the title compound, C16H13BrO, the two benzene rings are rotated in opposite directions with respect to the central C-C=C-C part of the molecule. The phenone O atom deviates from the least-squares plane of the molecule by 0.300 (3) A. In the crystal structure, molecules are paired through C H...pi interactions. The molecular pairs along [001] are hydrogen bonded through three translation-related co-operative hydrogen bonds in the 'bay area', forming molecular chains, which are further hydrogen bonded through C-H...Br weak interactions, forming (010) molecular layers. In the third direction, there are only weak van der Waals interactions. The co-operative hydrogen bonds in the 'bay area' are discussed briefly. PMID- 15345861 TI - Diethyl piperazine-1,4-diyldioxalate. AB - The ethyl oxamate group, N-C(O)-C(O)-OEt, in the title compound, alternatively called diethyl N,N':N,N'-bis(ethylene)dioxamate, C12H18N2O6, can be considered as being composed of two singly bonded amide and ester functionalities. The ethyl oxamate group is not planar. The two carbonyl groups are almost perpendicular, with an oxalyl O=C-C=O torsion angle of -111.34 (17) degrees. The molecule is located on an inversion centre. Infinite supramolecular tapes, propagating along the b axis, are formed through soft C-H...O interactions which form a centrosymmetric R(2)2(12) motif. PMID- 15345862 TI - N1-(4-tert-Butylphenyl)-N2-hydroxy-alpha-oxo-alpha-phenylacetamidine and N2 hydroxy-N1-(4-nitrophenyl)-alpha-oxo-alpha-phenylacetamidine hemihydrate. AB - In the title compounds, C18H20N2O2, (I), and C14H11N3O4.0.5H2O, (II), respectively, the oxime groups have an E configuration. In (I), the molecules exist as polymers bound by intermolecular C-H...O and O-H...N hydrogen bonds around inversion centres. In (II), intermolecular OW-H...N, OW-H...O and O-H...OW interactions stabilize the molecular packing. PMID- 15345863 TI - Two enamines derived from 1-n-alkyl-3-methylpyrazol-5-ones. AB - The first two crystal structures of enamines derived from 1-n-alkyl-3-methyl-5 pyrazolones, namely 1-(n-hexyl)-3-methyl-4-[1-(phenylamino)propylidene]-2 pyrazolin-5-one, C19H27N3O, (I), and N,N'-bis[1-[1-(n-hexyl)-3-methyl-5-oxo-2 pyrazolin-4-ylidene]ethyl]hexane-1,6-diamine, C30H52N6O2, (II), are reported. The molecule of (II) lies about an inversion centre. Both (I) and (II) are stabilized by intramolecular N-H...O hydrogen bonding. This confirms previous results based on spectroscopic evidence alone. PMID- 15345864 TI - AluElements: Repetitive DNA as Facilitators of Chromosomal Rearrangement. AB - Alu repeats are the most common type of repetitive DNA sequences dispersed throughout the human genome. Technical advances in the field of cytogenetics and molecular biology have facilitated the analysis of epithelial tumors and hematologic malignancies which has led to the observation of Alu elements in and near sites often involved in chromosomal rearrangements. Repair mechanisms of double strand breaks (DSB) such as homol-ogous recombination (HR) may rely on the sequence homology of Alu repeats, potentially leading to chromosomal rearrange ments. Databases have confirmed the strong association between Alu repeats, specifically the 26 bp consensus sequence and chro-mosomal regions involved in deletions and translocations. Although the Alu repetitive sequence is a potential "hotspot" during homologous recombination, there are other cellular mech-anisms that may play a more prominent role in the initiation of chromosomal rearrangements. PMID- 15345865 TI - Fanconi Anemia: A Decade of Discoveries. AB - In 1967, Guido Fanconi, a Swiss pediatrician, described Fanconi anemia (FA) in two siblings with similar physical anom-alies and bone marrow failure.1 A few years earlier, Schroeder and colleagues had described an increase in spontaneous chromosome breaks in blood samples from patients afflicted with what was then called familial panmyelopathy,2 now known to have been FA. Since then, the field of FA research has undergone an amazing revolution, especially in the last decade. Because of its relation-ship to DNA repair and association with a susceptibility to neo-plastic transformation, FA has gained much attention, particularly following the elucidation of some of the genes that are mutated in FA patients. In 2002, the discovery of a connection between FA, breast cancer, and other well-known DNA repair diseases sparked numerous publications on the subject. In this review, we discuss the clinical and genetic aspects of FA. PMID- 15345866 TI - Amnioinfusion in thick meconium. AB - OBJECTIVE: There are conflicting reports regarding the results of amnioinfusion in the management of meconium passage in utero. This study was done to evaluate transcervical amnioinfusion for meconium stained amniotic fluid during labour. METHODS: 196 women at term in early labour with meconium were randomized to receive either transcervical intrapartum amnioinfusion with saline (96) or routine obstetrical care (100). Transcervical amnioinfusion of one liter saline infused over 30-45 minutes. End points were relief of decelerations, incidence of vaginal delivery, presence of meconium below the neonatal cords, and X-ray evidence of meconium aspiration. RESULTS: Amnioinfusion resulted in relief of decelerations in 75% of cases as compared to 7% in the control group. Eighty eight percent of patients delivered vaginally as compared to 58% in the control group (p< 0.001). Neonatal outcome was significantly better in the infusion group. The incidence of meconium below the vocal cords was reduced from 48% to 17% (p< 0.004) using amnioinfusion with positive X-rays for meconium aspiration in only 12.5% versus 26% (p < 0.5). CONCLUSIONS: We concluded that transcervical intrapartum amnioinfusion is a safe, simple and inexpensive technique that reduces operative intervention and improves neonatal outcome, and is of tremendous relevance in developing countries. PMID- 15345867 TI - Fat-modified diets during pregnancy and lactation and serum lipids after birth. AB - OBJECTIVE: This study investigated the influence of modifying the maternal dietary fat on the serum lipids of infants at birth and at one year of age. METHODS: This single-blind randomized clinical trial was done on 180 4-month pregnant women. All subjects proved to have a fat-unmodified diet through a 4-day food record dietary questionnaire. They were divided randomly into two groups. The intervention group was kept on a fat-modified diet including saturated fatty acid (SFA) < 10%, monounsaturated fatty acids: (MUFA) 10-5%, polyunsaturated fatty acid (PUFA) upto 10% and cholesterol < 300 mg/day with dietary advice for the pregnancy period. The control group was given only the latter advice. All subjects were followed up monthly. The serum lipids including total cholesterol (T.cho), triglyceride (TG), and HDL cholesterol (HDL-C) were analyzed through enzymatic methods. The level of LDL-cholesterol (LDL-C) was calculated by Friedewald formula. The comparison of mean cord and one-year-old infant serum lipids were done through unpaired T-test in two groups. RESULTS: The mean level of T.cho in the intervention and control group was (70.3+/-15.9, vs 81.4+/-17.2, P< 0.009), TG (85.3 +/- 16.7 vs 97.5 +/- 18.2, P< 0.007), LDL-C (27.8 +/- 15.2 vs 34.8 +/- 17.1, P< 0.04) and non-HDL-C (44.5+/-7.2 vs 54.5 +/- 8.1, P< 0.02) and in one year old infant the comparison of serum lipids were as follows. T.cho (145.7 +/- 51.4 vs 161.4 +/- 56.2, P< 0.003), TG (90.1 +/- 31.8 vs 98.3 +/- 33.1, P< 0.02), LDL-C (85.6 +/- 20.4 vs 92.3 +/- 19.6, P< 0,05) and non-HDL-C (113.6 +/ 30.2 vs 128.8 +/- 34.8, P< 0.04). However, there was no significant difference in HDL-C of both groups. CONCLUSION: There is a significant decrease of T.cho, TG, LDL-C and non-HDL-C levels with no significant increase of HDL-C in the intervention group with the fat-modified diet. Maternal fat-modified diet could be suitable way to prevent cardiovascular disease among infants from the beginning of the life. PMID- 15345868 TI - Foreign body in upper digestive tract. AB - OBJECTIVE: Foreign body ingestion is a common clinical problem, encountered in children. METHODS: A total number of 104 patients with ingested foreign body admitted in our hospital are reviewed. Endoscopic removal was done for all foreign objects impacted in esophagus. RESULT: In 84.6% cases, history of having swallowed the foreign body was most common symptom. Majority of patients (61.53%) presented within 24 hours after ingestion of foreign body. Coins were the most frequent offending agents in children (87.5%). Seventy six cases of coins were impacted in the postcricoid region. Complications of retropharyngeal abscess was seen in two cases (1.92%), which was associated with sharp foreign body. CONCLUSION: Early removal of these sharp foreign bodies must be considered to reduce the risk of this complication. PMID- 15345869 TI - Psychopathology and coping in parents of chronically ill children. AB - OBJECTIVE: To evaluate psychopathology and coping mechanisms in parents of children with a chronic illness. METHODS: 30 parents whose children had Thalassaemia were randomly selected from the Thalassaemia Day Care Centre of a teaching general hospital. The parents were interviewed on a semi structured proforma and also rated on SCL-90-R and Mechanisms of Coping scales. The data was analyzed using Pearson's correlation coefficient. RESULTS: The parents were seen to have high psychopathology on SCL-90-R. Depression was the subscale with the maximum elevation. Somatisation was also significantly elevated The GSI (General Symptomatic Index), a measure of general distress was extremely high in about 86.66% of the parents. Most parents were fatalistic in their approach towards the illness. Fatalism positively correlated with somatisation and depression. Avoidant coping strategies were associated with higher scores on various dimensions of the SCL-90-R. CONCLUSION: Chronic illness in children affects the psychological health of the parents. Active coping strategies are associated with fewer distress indices and thus if inculcated may improve the ability to bear the burden of the illness without becoming themselves affected by psychiatric illnesses. PMID- 15345870 TI - Suicide among children and adolescents in South Delhi (1991-2000). AB - OBJECTIVE: Suicide among youth is of great concern and a subject requires thorough study to formulate prevention strategy. In this paper the incidence and trends of suicide among children and adolescent of South Delhi have been reported. METHODS: A retrospective analysis was carried out on 222 cases of suicidal deaths pertaining to age group of 10-18 years, the postmortem examination on the body of which were conducted in Department of Forensic Medicine and Toxicology, All India Institute of Medical Sciences, New Delhi during the period from 1st January 1991 to 31st December 2000. The particular of cases were analyzed according to age group, sex, method used and causes of committing suicide. RESULTS: Out of 222 cases 123(55.4%) were of girls (Female: Male 1.24:1). Commonest age group involved was 15-18 years in both the sexes. Commonest method used for committing suicide was hanging (57% in girls, 49.5% in boys) followed by poisoning (37.4% in girls, 49.5% in boys). CONCLUSION: Methods used to commit suicide are widely available and are difficult to restrict. Therefore, suicide prevention strategy based on risk factors could be more effective rather than limiting the access to methods. PMID- 15345871 TI - Control of Japanese encephalitis in India: a reality. AB - Japanese encephalitis is considered as a serious disease due to the complexity of the disease and lack of specific treatment. A secular trend towards declining of JE has been brought in China, Korea, and Japan with widespread use of JE vaccine. In India, the actual JE burden could be estimated only by strengthening diagnostic facilities for JE confirmation in hospitals. However, the available records at present indicate a rising trend in JE- occurrence and expansion of the disease into JE non-endemic areas, which cannot be ignored. JE control through vector control methods has limitations owing to sustainability and cost effectiveness of the programs. Under these circumstances, feasibility of JE vaccination in India has to be considered as a preventive measure, for which identification of risk areas, target populations to be immunized, cost-evaluation of immunization is emphasized. Since, JE vaccine is produced in India, extension of the availability of this vaccine into routine JE-immunization programs is not remote. China has proved that countries with limited sources can produce safe and effective JE vaccines. PMID- 15345872 TI - Kinase inhibitors translate lab discoveries into exciting new cures for cancers. AB - Genetic mutations can lead to abnormal activation of certain kinases that in turn lead to excessive cell division seen in cancers. Inhibitors of over activated kinases can theoretically inhibit cancer causing pathways and result in tumor shrinkage. These discoveries have sparked a revolution in drug discovery with many small molecule kinases inhibitors now being used in cancer clinical trials. The amazing success of Imatinib, a blocker of the bcr-abl kinase in chronic myeloid leukemia has shown that the drugs based on these strategies can improve cure rates in cancer. In this article, the authors review the concepts of kinase inhibition in cancer and principles behind the success of imitanib. The authors also review other promising kinase inhibitors being used in clinical trials that are expected to aid the fight against cancer. PMID- 15345874 TI - Newer drugs for asthma. AB - Several new drugs have resulted from innovative pharmacological and immunomodulatory approaches towards treatment of asthma. Diverse therapeutic targets have provided new agents. The new classes of drugs available are newer "on site activated inhaled corticisteroids" and soft steroids, anti IgE compounds, leukotriene inhibitors and single isomer agents which are available for clinical use. Anti interleukin agents and phosphodiesterase inhibitors are in the stage of clinical trials. Antisense therapy and pharmacogenetics are the on horizon for treatment of asthma. PMID- 15345875 TI - Management of unresponsive asthma. AB - Difficult asthma is defined as asthma that is not controlled despite treatment with> 800 micro g budesonide or equivalent per day. Poor control is defined as the need for bronchodilators more than three times a week, school absence of more than five days a term, or one episode or more of wheezing each month. Common causes of poor response to treatment include; wrong diagnosis, inappropriate medications or improper inhalation technique, poor adherence to medications and co-morbidity. Steroid resistant asthma is uncommon and estimated to be 1 in 1000 10000 asthmatic patients. If there is no functional improvement to prednisolone 2 mg/kg/day for 2 weeks with adherence checked by measuring serum prednisolone and cortisol levels, a fibreoptic bronchoscopic examination with bronchoalveolar lavage and large airway biopsy should be considered. Eosinophilic inflammation identified on the biopsy in a child who is unresponsive to prednisolone may benefit from alternative anti-inflammatory treatments such as cyclosporin. Neutrophilic infiltration in biopsy may benefit with macrolide antibiotics, 5 lipogenase inhibitors or theophyllines. PMID- 15345876 TI - Evidence based treatment of bronchiolitis. PMID- 15345877 TI - Bronchoscopist talks to you. AB - Fiberoptic bronchoscopy (FBS) is an important entry in the armamentarium of procedures listed in management of respiratory problems. It is a simple and a safe procedure. FBS has a great scope for diagnosis as well as therapy in pediatric respiratory illnesses. This article gives a practical overview of FBS in pediatric practice. PMID- 15345878 TI - Acute lung injury and acute respiratory distress syndrome. AB - Acute lung injury and acute respiratory distress syndrome are an important challenge for pediatric intensive care units. These disorders are characterized by a significant inflammatory response to a local (pulmonary) or remote (systemic) insult resulting in injury to alveolar epithelial and endothelial barriers of the lung, acute inflammation and protein rich pulmonary edema. The reported rates in children vary from 8.5 to 16 cases / 1000 pediatric intensive care unit (PICU) admissions. The pathological features of ARDS are described as passing through three overlapping phases - an inflammatory or exudative phase (0 7 days), a proliferative phase (7-21 days) and lastly a fibrotic phase (from day 10). The treatment of ARDS rests on good supportive care and control of initiating cause. The goal of ventilating patients with ALI/ARDS should be to maintain adequate gas exchange with minimal ventilator induced lung injury. This can be achieved by use of optimum PEEP, low tidal volume and appropriate FiO2. High frequency ventilation can improve oxygenation but does affect the outcomes. Prone positioning is a useful strategy to improve oxygenation. Pharmacological strategies have not made any significant impact on the outcomes. Preliminary data suggests some role for use of corticosteroids in non-resolving ARDS. The mortality rates have declined over the last decade chiefly due to the advances in supporting critically ill patients. PMID- 15345879 TI - Unilateral internal jugular phlebectasia. AB - Jugular phlebectasia is a rare cause of cervical swelling in children. It is a fusiform dilatation of any part of the jugular venous system and can involve the external, internal or anterior jugular veins. Previous reports suggest that the entity is often ignored or misdiagnosed. Unilateral internal jugular phlebactasia presenting as an intermittent neck swelling in a ten-year-old girl is reported. The clinical features are analyzed and the appropriate use of noninvasive imaging modalities is highlighted. The literature is also briefly reviewed. PMID- 15345880 TI - Separation of thoraco-omphalopagus Siamese twin. AB - A pair of female thoraco-omphalopagus twins, with two pairs of lungs, common diaphragm and separate pleural cavities was separated at the age of 5(1/2) months after a parasitic relationship had developed between them. Before separation both the babies developed recurrent respiratory tract infection and frequent diarrhea. They were treated with medical therapy and made fit for anesthesia. The combined weight of the twins was 4(1/2) kg at birth and the combined weight at the time of separation was 9(1/2) kg. One baby was smaller in growth compared to the other baby. The closure of anterior abdominal wall was difficult in both babies. However, we could close it without use of any biodegradable patches or tissue expander. Both the babies survived and are doing well after 3 months of separation. PMID- 15345881 TI - Bjornstad syndrome. AB - A 9-month-old boy presented with the complaints of loose motion, vomiting and difficulty in breathing. His scalp hairs were thin, brittle, and sparse and were of differing lengths with twisted appearance. Hair shaft microscopy revealed alternate light and dark segments and twisting of the hair shafts by 180 degrees along the axis. Serum copper levels were normal. The audiological testing revealed bilateral sensorineural hearing loss. Child was diagnosed as a case of Bjornstad Syndrome. PMID- 15345882 TI - Choroid plexus papilloma associated with developmental delay. AB - Choroid plexus papillomas are rare, benign tumors of neuroectodermal origin usually observed in the lateral ventricles of children. The usual presenting signs of choroid plexus papillomas are related to hydrocephalus and increased intracranial pressure. A child presented to us with clinical features of delayed milestones, which was later diagnosed as a case of choroid plexus papilloma with hydrocephalus. He underwent complete excision of the tumour with gradual recovery of milestones. PMID- 15345883 TI - Typhoid vaccination after enteric fever. PMID- 15345885 TI - Health information on the internet: patient empowerment or patient deceit? AB - Internet is the biggest medical library in the world. It has transformed the way many health seekers find health information. Seekers on net have exponentially increased from 54 million in 1998 to 110 million in 2002 (U.S. figures) and are ever increasing. Act of looking for health or medical information is the third most popular activities online. Search engines are used by almost 81% of the e patients to look for the information they want. Internet is fast becoming an influential force as more than 70% consumers say that the information on the net has influenced their treatment decisions. However, the reliability of all the health information available on the Internet is questionable. Numerous studies have indicated deficiencies in the quality of information on the Internet. Due to technical and commercial reasons, the results provided by various search engines can be potentially biased. Only about a quarter of health seekers thoroughly check the source, timeliness of information every time they search for health information. In fact, most adults from USA, Japan, France and Germany who participated in a recent survey thought online health care information to be trustworthy, of good quality, easy to understand and easy to find. Efforts are now being taken to ensure the quality of health information on the Internet. Patients need to be educated about the worthiness of a site and also be prescribed the right sites to be consulted for information. PMID- 15345886 TI - Characteristics of acute bacterial meningitis in Southeast Turkey. AB - BACKGROUND: Acute bacterial meningitis (ABM) remains a significant worldwide cause of death in adults. Even in the antibiotic era, the mortality rate in ABM remains significant and has been reported in the range of 8-40%. AIM: The aim of this study was to assess the characteristics of epidemiology, clinical manifestations, treatment modalities and outcome of patients with ABM in Southeast Turkey. SETTINGS AND DESIGN: This retrospective study included all cases of community-acquired ABM diagnosed and treated in Dicle University Hospital between June 1996 and December 2002. METHODS AND PATIENTS: The study group consisted of 186 adult patients (110 male, 76 female) with ABM, those patients who are older than 14 years, followed up at Dicle University Hospital from June 1996 to December 2002. Patients' charts were retrospectively reviewed, clinical characteristics were recorded and final data were analyzed. STATISTICAL ANALYSIS: In statistical analyses, the Chi-square test was used for binary variables and Student's t-test for continuous variables. RESULTS: The patients' mean age was 30.2 +/- 15.3 years (range 14 to 90 years). On admission, typical symptoms of meningitis were found in most of the patients: headache in 92.5%, fever in 88.2%, and nuchal rigidity in 80.1%. The main predisposing factor for ABM was otitis media (40 patients, 21.5%) and closed head trauma (12 patients, 6.5%). Streptococcus pneumonia was the most common identified pathogen. Twenty nine patients (15.6%) died during hospitalization period. In multivariate analyses, the significant mortality factor was found as initial level of consciousness, low cerebrospinal fluid/blood glucose ratio, high erythrocyte sedimentation rate and initial treatment by penicillin G. CONCLUSIONS: Although still remains as a serious infection, early diagnosis and effective treatment may reduce fatal outcome and improve the course of the disease in patients with ABM. Ceftriaxone should be considered as the drug of choice for initial empirical therapy, while waiting culture results and vancomycin must be withheld for patients having increased risk of penicillin resistant pneumococci strains. PMID- 15345887 TI - Injection practices in a metropolis of North India: perceptions, determinants and issues of safety. AB - BACKGROUND: At least 50 percent of the injections administered each year are unsafe, more particularly in developing countries, posing serious health risks. An initial assessment to describe injection practices; their determinants and adverse effects can prevent injection-associated transmission of blood borne pathogens by reducing injection frequency and adoption of safe injection practices. AIMS: To assess the injection practices in a large metropolitan city encompassing varied socio-cultural scenarios. STUDY SETTING AND DESIGN: Field based cross sectional survey covering urban non-slum, slum and peri-urban areas of a large metropolitan city. METHODS AND MATERIAL: Injection prescribers, providers and community members selected by random sampling from the study areas. Pre tested questionnaires assessed knowledge and perceptions of study subjects towards injections and their possible complications. Observation of the process of injection and prescription audit also carried out. STATISTICAL ANALYSIS: MS Access for database and SPSS ver 11 for analysis. Point estimates, 95% confidence intervals, Chi Square, t test, one-way ANOVA. RESULTS: The per capita injection rate was 5.1 per year and ratio of therapeutic to immunization injections was 4.4:1. Only 22.5%of injections were administered with a sterile syringe and needle. The level of knowledge about HIV and HBV transmission by unsafe injections was satisfactory amongst prescribers and community, but inadequate amongst providers. HCV was known to a very few in all the groups. The annual incidence of needle stick injuries among providers was quite high. CONCLUSION: A locally relevant safe injection policy based on multi disciplinary approach is required to reduce number of injections, unsafe injections and their attendant complications. PMID- 15345888 TI - Role of myeloperoxidase index in differentiation of megaloblastic and aplastic anemia. AB - BACKGROUND: Elevated neutrophil myeloperoxidase may have a role in the diagnosis of megaloblastic erythropoiesis. AIMS: To study the differentiating role of myeloperoxidase index in megaloblastic and aplastic anemia. SETTINGS AND DESIGN: The myeloperoxidase index (MPXI) was studied in 96 patients with megaloblastic and aplastic anemia diagnosed on bone marrow aspiration and biopsy examinations. METHODS AND MATERIALS: MPXI was measured with Technicon H1 (Bayer) automated analyzer. Nonparametric Mann-Whitney statistical test was used to compare the MPXI values between groups. RESULTS: The mean MPXI in megaloblastics and aplastic anemia was 18.3 and 1.8 (p< 0.001) respectively. MPXI> 20 denoted megaloblastic and MPXI <-11.6 denoted aplastic anemia. CONCLUSION: MPXI measurement may assist differentiation of megaloblastic from aplastic anemia, while MPXI> 20 rules out aplastic and MPXI <-11.6 rules out megaloblastic anemia. PMID- 15345889 TI - Concomitant gout and rheumatoid arthritis--a case report. AB - We report a case of definite rheumatoid arthritis and co-existing gout. Although gout and rheumatoid arthritis are relatively common entities individually, the co existence of these two conditions is rare. PMID- 15345890 TI - Pulmonary artery catheter insertion in a patient of dextrocardia with anomalous venous connections. AB - In a young adult patient having situs solitus with dextrocardia the attempted pulmonary artery catheter placement for emergency mitral valve replacement required an unduly long length (50cm) of catheter insertion to get into right ventricle and then into pulmonary artery. Although catheter coiling was suspected initially, chest x-ray taken after successfully placement revealed an uncommon congenital anomalous venous connection i.e. right internal jugular opening into left sided superior vena cava then into inferior vena cava after running all along the left border of the heart. With the result, it required to pass 50cm of PA catheter to get into right ventricle in our patient. This emphasizes the need to look for abnormal venous connections during echocardiography and x-ray screening in congenital heart disease. Fluoroscopy is recommended when an unusual length of pulmonary artery catheter insertion is required to enter the pulmonary artery. PMID- 15345891 TI - Judicial activism for making operational all anganwadi centres in India. PMID- 15345892 TI - Topotecan, Ara-C, cisplatin and prednisolone (TOPOSHAP) for patients with refractory and relapsing lymphomas: results of a phase I trial. AB - We designed a phase I trial to assess the feasibility of the combination of topotecan, Ara-C, cisplatin and solumedrol (TOPOSHAP) in patients with relapsed or primary refractory lymphomas. We included 9 patients with measurable non Hodgkin's (n = 8) and Hodgkin's (n = 1) lymphomas. Level 1 consisted of topotecan 1.0 mg/m(2)/day, i.v., given on days 1-3, cisplatin 25 mg/m(2)/day, i.v., on days 1-3, Ara-C 500 mg/m(2), i.v., on day 4, methylprednisolone 250 mg, i.v., on days 1-4. The regimen was repeated every 3-4 weeks. The maximum tolerated dose was already reached at level 1. G-CSF was added systematically after the 5th patient was included. The most significant toxicity in this trial was hematologic (all had neutropenia WHO grade 4 and 7 had grade 4 thrombocytopenia). Three patients had neutropenic fever. We observed two instances of WHO grade 3 and one of grade 4 diarrhea. Two patients achieved a complete response and 6 a partial response. We conclude that TOPOSHAP with G-CSF support is feasible and should be further studied in phase II studies. PMID- 15345893 TI - Effect of iron deficiency anemia on the levels of hemoglobin A1c in nondiabetic patients. AB - The major form of glycohemoglobin is hemoglobin A1c (HbA1c). The HbA1c fraction is abnormally elevated in chronic hyperglycemic diabetic patients and correlates positively with glycemic control. Previous studies suggest that iron deficiency anemia (IDA) affects the levels of HbA1c. The aim of this study was to determine the effect of IDA on HbA1c levels in nondiabetic patients. The population studied consisted of 50 patients (30 women, 20 men, mean age 35.7 +/- 11.9 years) with IDA and 50 healthy subjects that were matched for age and sex. Patients who had glucose tolerance abnormalities (impaired glucose tolerance or diabetes mellitus), hemoglobinopathies, hemolytic anemia, chronic alcohol ingestion and chronic renal failure were excluded from the study. Hematologic investigations, fasting and postprandial glucose and HbA1c levels were measured in all subjects before iron therapy. All patients with IDA were treated with iron 100 mg/day for 3 months. We repeated the laboratory investigation after iron therapy. Before iron treatment, the mean HbA1c (7.4 +/- 0.8%) level in patients with IDA was higher than in a healthy group (5.9% +/- 0.5) (p < 0.001). In patients with IDA, HbA1c decreased significantly after iron treatment from a mean of 7.4% +/- 0.8 to 6.2% +/- 0.6 (p < 0.001). Iron deficiency must be corrected before any diagnostic or therapeutic decision is made based on HbA1c. PMID- 15345894 TI - Lymphocyte populations in lymph nodes in different histological types of Hodgkin's disease in children. AB - Proliferating Reed-Sternberg and Hodgkin's cells within the lymph node in Hodgkin's disease (HD) affect the surrounding cells, which results in alterations of the proportion between T and B lymphocytes and the activation of these cells. The purpose of this study was to assay the changes within lymphocyte populations in lymph nodes in different types of HD. The lymph nodes of 10 children with nonmalignant, reactive changes, and 31 children with HD of the following types: mixed cellularity (MC; 11 cases), nodular sclerosis (NS; 17 cases) and lymphocyte depletion (LD; 3 cases) were included in the study. The percentage of T lymphocytes (CD3+), B lymphocytes (CD22+), T cell (CD4+ and CD8+) and B cell subsets (kappa+ and lambda+ light chain) were assayed with flow cytometry. The ratio of T:B lymphocytes in reactive lymph nodes and in NS type of HD was similar. An increase of the T cell percentage (resembling an increase of the CD4+ subset) and a decrease of B cells resulted in a high ratio of T:B and was noted in the MC and LD types of HD. These HD types were associated with marked changes in lymphocytes within the lymph node, which suggested a different effect of proliferating Reed-Sternberg and Hodgkin cells in HD on the surrounding cells. PMID- 15345895 TI - Amifostine stimulates the formation of hematopoietic bone marrow progenitors from B-cell chronic lymphocytic leukemia. AB - Amifostine is a phosphorylated aminothiol that not only protects hematopoietic progenitor cells from chemotherapy and radiotherapy, but also stimulates normal hematopoiesis. The effect of amifostine on the in vitro growth of hematopoietic progenitors derived from B-cell chronic lymphocytic leukemia(B-CLL) was investigated. The colony-forming units (CFU)-granulocyte macrophage (CFU-GM), the burst-forming units-erythroid (BFU-E) and the CFU-granulocyte erythroid macrophage megakaryocytes (CFU-GEMM) increased 38, 20 and 100%, respectively, after the incubation with amifostine. There was no statistical difference in the in vitro progenitor growth of patients grouped according to their disease stage, bone marrow lymphocytic infiltration or therapy. Our data indicate that apart from cytoprotection the parallel use of amifostine and chemotherapy in patients with B-CLL could enhance bone marrow recovery. PMID- 15345896 TI - Stanford V regimen plus consolidative radiotherapy is an effective therapeutic program for bulky or advanced-stage Hodgkin's disease. AB - Since September 1996, 48 untreated patients with bulky or advanced-stage Hodgkin's disease received the 12-week Stanford V chemotherapy regimen followed by consolidation radiotherapy at a dose of 36 Gy to bulky mediastinal disease and 30.6 Gy to the initial sites of disease > or =3 cm in transverse diameter. After the combined therapy, 46 of 48 (96%) achieved complete remissions. With a median follow-up of 48 months, the 5-year overall survival was 95% and freedom from progression 86%. There were no treatment-related deaths. All but one premenopausal female patient (who received pelvic and inguinal irradiation) recovered normal menses. Until now no case of secondary leukemia or myelodysplasia was observed. Our results confirm that the Stanford V regimen with consolidation radiotherapy is safe and effective in patients with bulky or advanced-stage Hodgkin's disease, achieving very high remission and overall 5 year survival rates. Longer follow-up is necessary to evaluate the extent of all complications. PMID- 15345897 TI - Is the treatment of Hodgkin's disease detrimental to the parathyroid gland? AB - The thyroid gland is often injured by supradiaphragmatic irradiation for Hodgkin's lymphoma. The aim of the present study was to examine whether the parathyroid gland gets injured by the treatment for Hodgkin's disease. Calcium, phosphorus and parathormone levels of 143 patients with primary treatment for Hodgkin's disease and in complete remission for 2 years were measured as well as the presence of antiparathyroid antibody in patients having antithyroid antibody. Out of the 143 patients studied, 104 received neck irradiation (with or without chemotherapy); among them laboratory alterations were observed in 7 cases. 39 patients received only chemotherapy; 3 of them had alterations. In contrast to the injury of the thyroid gland, no damage to the parathyroid glands associated with the treatment for Hodgkin's disease was noted. It has been concluded that the use of high-dose external radiotherapy does not mean a higher risk as regards the parathyroid gland but further follow-up studies of the patients may result in the revelation of the development of parathyroid lesions. PMID- 15345898 TI - Analysis of the optimal blood sampling conditions for estimation of hematopoietic progenitor cell count by the SE-9000 automated hematolyzer. AB - We evaluated the optimal conditions for blood sampling for hematopoietic progenitor cells (HPCs) as estimated by the immature information program of the SE-9000 automated hematology analyzer. The HPC count was most stable when the blood samples were incubated at room temperature with ethylene-diaminetetraacetic acid dipotassium (EDTA-2K) as an anticoagulant. The HPC count should, however, be measured within 4 h after blood collection, even under optimal conditions. In contrast, the CD34+ cell count estimated by flow cytometric analysis was stable for at least 21 h after the blood samples were incubated with EDTA-2K at room temperature or 4 degrees C. When appropriate blood samples were used, the HPC count in the peripheral blood significantly correlated with the CD34+ cell count in the peripheral blood and in the apheresis yields (r = 0.798 and 0.635, respectively); therefore, the HPC count is a reliable predictor for initiation of apheresis procedures to obtain sufficient HPCs for peripheral blood stem cell transplantation. PMID- 15345899 TI - Severe hypercalcemia in a child with acute nonlymphocytic leukemia: the role of parathyroid hormone-related protein and proinflammatory cytokines. AB - Among the hematological malignancies, hypercalcemia has often been reported in lymphoid malignancies such as multiple myeloma and adult T cell leukemia/lymphoma, but it has only rarely been described in acute nonlymphocytic leukemia. We describe here a 14-month-old girl with acute monocytic leukemia complicated by severe hypercalcemia (4.6 mmol/l) at presentation. A bone survey showed generalized bone resorption, but no localized osteolytic lesions. A search for the etiology of the hypercalcemia revealed that the serum levels of parathyroid hormone-related protein (PTHrP) and also proinflammatory cytokines with stimulatory effects on osteolytic bone resorption - TNF-alpha, IL-6 and M CSF - were elevated. The patient achieved complete remission with induction chemotherapy, and the levels of PTHrP and the cytokines became normalized. In this case, PTHrP and cytokines might have acted cooperatively to exacerbate bone resorption, resulting in severe hypercalcemia. PMID- 15345900 TI - Eosinophilic gastroenteritis or eosinophilic chloroma? AB - Granulocytic sarcoma of the small intestine preceding or as the presenting feature of acute myelogenous leukemia with chromosome 16 abnormalities has been observed in at least 4 patients. We report the case of a patient initially diagnosed with eosinophilic gastroenteritis, responding to corticosteroid treatment for 21 months and eventually transforming into an acute myelogenous leukemia with eosinophilia (M4Eo variant). PMID- 15345901 TI - Transient pancytopenia preceding T lineage acute lymphoblastic leukemia. AB - Transient pancytopenia preceding acute lymphoblastic leukemia (pre-ALL) is a rare occurrence usually affecting children with subsequent development of B lineage ALL. We report a case of pre-ALL characterized by a T cell immunophenotype and abnormal karyotype t (11; 14) (q10; q10). The patient achieved a transient complete remission after initial therapy, but relapsed within a few months and died of leukemic encephalopathy. To the best of our knowledge, this is the first report of T lineage pre-ALL. PMID- 15345902 TI - Serum basic fibroblast growth factor and vascular endothelial growth factor levels are related in patients with polycythaemia vera. PMID- 15345903 TI - Uptake of magnetic beads by dendritic cells, a major obstacle for the use of beads in biological assays. PMID- 15345904 TI - Reported case of severe hepatotoxicity likely due to fluconazole and not desloratadine. PMID- 15345905 TI - Bisphenol A exerts thyroid-hormone-like effects on mouse oligodendrocyte precursor cells. AB - We report studies on the mechanism of action of bisphenol A (BPA) on the differentiation of oligodendrocyte precursor cells (OPCs). Our results show that: (1) BPA inhibits the differentiation of OPCs induced by exposure to thyroid hormone (T3). (2) The effect is mediated through various mechanisms via the thyroid hormone receptor (TRbeta1) which is considered to be responsible for OPC differentiation. (3) The action of BPA on OPC differentiation does not involve the FcRgamma-Fyn-myelin basic protein (MBP) cascade as an inducer of OPC differentiation nor does it suppress CREB phosphorylation, which is considered to be induced by the T3-TR complex. (4) The presence of MBP isoforms (21.5, 18.5, 17.0 and 14.0 kDa) was detected in OPCs, and the expression of exon 2-containing isoforms (i.e. 17.0 and 21.5 kDa) was upregulated upon treatment with T3. In contrast, expression of MBP was inhibited by BPA. PMID- 15345906 TI - Social hierarchy affects gene expression for catecholamine biosynthetic enzymes in rat adrenal glands. AB - Social stressors, like other stressors, are powerful activators of the sympathoadrenomedullary system. Differential housing (single vs. group) and social defeat of rats is known to alter the activity of catecholamine synthesizing enzymes in the medulla. The present studies examined the effect of 70 days of triad (3 rats per large cage) and individual housing of male rats on adrenal mRNA levels of tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT) and on TH protein levels. Behavioral ratings carried out at the triad formation indicated that dominant rats exhibited mostly offensive aggressive behaviors. By contrast, subordinate rats expressed primarily defensive behaviors, while the subdominant rats displayed intermediate levels of these behaviors. Overall, compared with single housing, triad housing resulted in lower gene expression for TH, DBH and PNMT and lower TH protein in the adrenals. Within triads, gene expression for these enzymes and TH protein concentration were higher in subordinate compared with dominant and subdominant rats. The dominant rats tended to have the lowest gene expression of these enzymes. These data indicate that in rodents, individual housing and a subject's social rank have a differential impact on the regulation of catecholamine biosynthesis already during the process of gene expression of catecholamine biosynthetic enzymes in the adrenals. PMID- 15345907 TI - Elevated levels of IgG and IgG4 to Malassezia allergens in atopic eczema patients with IgE reactivity to Malassezia. AB - BACKGROUND: The opportunistic yeast Malassezia is considered to be one of the factors that can contribute to atopic eczema (AE). Elevated serum IgE levels, T cell proliferation and positive skin prick test (SPT) and atopy patch test (APT) reactions to Malassezia are found among AE patients. METHODS: Sera from 127 AE patients, 14 patients with seborrheic dermatitis (SD) and 33 healthy controls were investigated for IgE and IgG4 to M. sympodialis extract and four recombinant Malassezia allergens; rMala s 1, rMala s 5, rMala s 6, and rMala s 9. In addition, IgG to the recombinant allergens was analyzed. The IgG and IgG4 levels were compared to IgE levels and in vivo reactions (SPT and APT) to Malassezia. RESULTS: AE patients with serum IgE levels >0.35 kU/l to M. sympodialis extract had significantly higher IgG4 levels to M. sympodialis extract than AE patients without detectable serum IgE to M. sympodialis extract, SD patients and healthy controls. Among the AE patients with and without detectable serum IgE to M. sympodialis extract, respectively, there were no differences in IgG4 levels between patients with positive or negative in vivo reactions to M. sympodialis extract. IgG4 to the rMala s allergens was almost exclusively found among patients with IgE to the same allergen. Within the four tested rMala s allergens, most IgG4 reactions were found to rMala s 6, an allergen with homology to cyclophilin. CONCLUSIONS: Elevated serum IgG4 to M. sympodialis extract accompanies elevated serum IgE to the extract. This is further confirmed by the association between IgG/IgG4 and IgE to recombinant Malassezia allergens. PMID- 15345908 TI - Isoforms of the major peanut allergen Ara h 2: IgE binding in children with peanut allergy. AB - BACKGROUND: The major peanut allergen Ara h 2 consists of two isoforms, namely Ara h 2.0101 and Ara h 2.0201. The recently identified Ara h 2.0201 isoform contains an extra 12 amino acids including an extra copy of the reported immunodominant epitope DPYSPS. This study aimed to evaluate the IgE binding of the two Ara h 2 isoforms. METHODS: Ten clones of Ara h 2 were sequenced to assess the relative frequency of the Ara h 2 isoforms and to identify whether there was further variation in the Ara h 2 sequence. IgE binding to Ara h 2.0101 and Ara h 2.0201 was measured for 70 peanut-allergic children using an IgE DELFIA assay to quantitate specific IgE binding. A competition assay was used to measure whether Ara h 2.0201 contained IgE epitopes other than those found for Ara h 2.0101. RESULTS: The original Ara h 2.0101 sequence was found for 6/10 clones and Ara h 2.0201 was found for 2/10 clones. Ara h 2.0201 had the expected insertion of 12 amino acids as well as substitutions at positions 40 (40G) and 142 (142E). Two new isoforms were identified as different polymorphisms of position 142. One Ara h 2.01 clone (Ara h 2.0102) contained 142E and one Ara h 2.02 clone (Ara h 2.0202) contained 142D. A polymorphism that was previously identified by other investigators at position 77 (77Q or 77R) was not found for any of the 10 sequences. Although the level of IgE binding to Ara h 2.0201 of individual patients was frequently higher than the binding to Ara h 2.0101 (p < 0.01), there was a strong correlation in binding to both isoforms (r = 0.987, p < 0.0001) and when analyzed as a group the means were similar. Ara h 2.0101 was not as efficient at blocking reactivity to Ara h 2.0201 indicating there is an additional IgE specificity for the Ara h 2.0201 isoform. CONCLUSIONS: Ara h 2.0201 has similar but higher IgE binding than the originally sequenced Ara h 2.0101 isoform and contains other IgE specificities. PMID- 15345909 TI - Food allergens are protected from degradation during CD23-mediated transepithelial transport. AB - BACKGROUND: CD23 (FcepsilonRII) is expressed by intestinal epithelial cells (IEC) following allergic stimulation and increases the uptake of IgE/allergen complexes. The aim of this study was to further analyze the role of CD23 in the intraepithelial processing of food allergens during transepithelial transport. METHODS: Balb-C mice were sensitized intraperitoneally with horseradish peroxidase (HRP) or beta-lactoglobulin (beta-LG) in the presence of pertussis toxin. In control and sensitized mice, 3H-HRP, intact HRP, or 14C-beta-LG fluxes were measured across jejunal segments mounted in Ussing chambers, in the presence or absence of mucosal anti-CD23 antibodies. HPLC analysis of serosal buffer was performed to detect HRP- or beta-LG-derived radiolabelled metabolites generated during transepithelial transport. RESULTS: In HRP-sensitized mice, 3H-HRP fluxes and intact HRP fluxes (3,836 +/- 476 and 290 +/- 86 ng/h x cm2, respectively) were significantly increased compared to control mice (1,677 +/- 297 ng/h x cm2, p < 0.01, and 106 +/- 23 ng/h x cm2, p < 0.02, respectively). HPLC analysis indicated the presence of intact HRP in the serosal compartment already 10 min after addition of HRP to the mucosal compartment, a result not observed in the control mice. In the presence of anti-CD23 antibodies, intact HRP fluxes were significantly decreased (131 +/- 27 ng/h x cm2) compared to control values in sensitized mice (290 +/- 86 ng/h x cm2, p < 0.02), suggesting that CD23 is involved is this 'protected' transport pathway. A similar protection during intestinal transport was observed for beta-LG in beta-LG sensitized mice. CONCLUSIONS: These results confirm that CD23 is involved in the rapid transepithelial transport of intact allergens in sensitized animals, and indicate that CD23 opens a 'protected' transport pathway in IECs. PMID- 15345910 TI - An experimental model of allergic asthma in cats sensitized to house dust mite or bermuda grass allergen. AB - BACKGROUND: Animal models are used to mimic human asthma, however, not all models replicate the major characteristics of the human disease. Spontaneous development of asthma with hallmark features similar to humans has been documented to occur with relative frequency in only one animal species, the cat. We hypothesized that we could develop an experimental model of feline asthma using clinically relevant aeroallergens identified from cases of naturally developing feline asthma, and characterize immunologic, physiologic, and pathologic changes over 1 year. METHODS: House dust mite (HDMA) and Bermuda grass (BGA) allergen were selected by screening 10 privately owned pet cats with spontaneous asthma using a serum allergen-specific IgE ELISA. Parenteral sensitization and aerosol challenges were used to replicate the naturally developing disease in research cats. The asthmatic phenotype was characterized using intradermal skin testing, serum allergen-specific IgE ELISA, serum and bronchoalveolar lavage fluid (BALF) IgG and IgA ELISAs, airway hyperresponsiveness testing, BALF cytology, cytokine profiles using TaqMan PCR, and histopathologic evaluation. RESULTS: Sensitization with HDMA or BGA in cats led to allergen-specific IgE production, allergen specific serum and BALF IgG and IgA production, airway hyperreactivity, airway eosinophilia, an acute T helper 2 cytokine profile in peripheral blood mononuclear cells and BALF cells, and histologic evidence of airway remodeling. CONCLUSIONS: Using clinically relevant aeroallergens to sensitize and challenge the cat provides an additional animal model to study the immunopathophysiologic mechanisms of allergic asthma. Chronic exposure to allergen in the cat leads to a variety of immunologic, physiologic, and pathologic changes that mimic the features seen in human asthma. PMID- 15345911 TI - Dietary nucleic acid and intestinal microbiota synergistically promote a shift in the Th1/Th2 balance toward Th1-skewed immunity. AB - BACKGROUND: Intestinal microbiota are known to play an important role in the establishment of oral tolerance, thereby protecting the organism from food allergies. Dietary intake of nucleic acid (NA) is also reported to have such an anti-allergic effect; however, one unsolved question is whether or not dietary NA would act through a process of toll-like receptor 9 signaling activated by DNA containing a CpG motif, a well-known sequence leading to immunostimulatory activity. In this study, we focused on the question of whether the addition of dietary NA lacking CpG motifs would allow continued modulation of the Th1/Th2 balance. METHODS: Germ free (GF) and Bifidobacterium-infantis-monoassociated BALB/c mice were maintained on either an NA-free casein diet or on an NA supplemented casein diet for 4 weeks. Thereafter, both the in vivo anti-casein antibody levels and in vitro splenocyte cytokine secretion pattern were evaluated. RESULTS: Feeding with a casein diet elicited a substantial increase in the serum anti-casein-specific IgG1, IgG2a, and IgE levels of GF mice fed the NA free-diet. The in vitro cytokine production profile showed that enhanced IL-4 production in the GF mice fed the NA free-diet was markedly reduced by the supplementation with dietary NA in both the GF and B.-infantis-monoassociated mice. In addition, IFN-gamma secretion increased in the B.-infantis-reconstituted mice fed the diet containing NA. CONCLUSIONS: These results suggest that dietary intake of NA devoid of CpG motifs may prevent the development of allergies via acceleration of Th1-dominant immunity. PMID- 15345912 TI - Skin reactivity to histamine and codeine in unselected 9-year-old children from Italy, Poland and Libya. AB - BACKGROUND: Previous studies have shown that histamine skin reactivity (the dimensions of a skin wheal elicited by a prick with histamine 10 mg/ml) in unselected school children has increased in Italy during the past two decades and is higher in Italy than in Poland. Hence this variable can probably be influenced by a changing or different lifestyle. The aim of this study was to compare skin reactivity to histamine and codeine (a marker of histamine releasability from mast cells) in schoolchildren from countries with different lifestyles. METHODS: Six previously unstudied unselected populations of 9-year-old schoolchildren (two each from Poland, Italy, and Libya; n = 863 subjects; 49.0% males) were pricked with two concentrations of histamine (10 and 1 mg/ml) and codeine (90 and 9 mg/ml). RESULTS: The higher concentrations of both pharmacologic agents tested yielded significantly different wheal areas in the three countries: Poland < Italy < Libya (histamine, 11.8, 16.1 and 20.7 mm2; codeine, 9.2, 13.2 and 16.2 mm2; p < 0.001 for all comparisons). The lower concentrations elicited almost matching results. Histamine wheal areas correlated closely with areas elicited by codeine in the same individual: angular coefficients of the histamine to codeine regression lines were 0.535, Italy; 0.551, Libya; 0.612, Poland; and 0.581 for the whole population. More histamine was needed to produce a wheal in Poland than in Libya: a 20-mm2 wheal required an injected histamine concentration of about 8.8 mg/ml in Libya, 29.5 mg/ml in Italy and 102.1 mg/ml in Poland. CONCLUSION: More studies are necessary to explain the observed international differences in skin histamine reactivity and their effect on the prevalence of positive allergen skin tests. PMID- 15345913 TI - Comparison of the effects in the nose and skin of a single dose of desloratadine and levocetirizine over 24 hours. AB - BACKGROUND: Desloratadine (DL) and levocetirizine (LCZ) are the newest commercialized antihistamines. Pharmacokinetics, pharmacodynamics and clinical data are available for both drugs, but there is to date no direct comparison involving the nose and skin at the same time. We compared the effects of a single dose of the two drugs in the nose and skin over 24 h. METHODS: Twenty-three patients with symptomatic allergic rhinitis were enrolled in a randomized double blind crossover administration of DL and LCZ. The histamine-induced wheal and flare was measured at baseline and 2 and 24 h after dosing. A reflective total symptom score (rTSS) for the previous 24 h was assessed before and after each dose. An instant symptom score was also measured at various time points after each drug. RESULTS: LCZ provided greater inhibition of the flare at 2 h (p = 0.05) and at 24 h (p = 0.007) and greater inhibition of the wheal only at 2 h (p = 0.02). The decrease in wheal and flare was significant versus baseline (p = 0.007) with both drugs. The rTSS of the previous 24 h decreased significantly with both LCZ (11.53 vs. 8.0; p < 0.05) and DL (11.3 vs. 7.9; p < 0.05). The instant TSS progressively decreased in parallel with both drugs, but a difference in favor of LCZ was seen 2 h after dosing. CONCLUSIONS: Single doses of DL and LCZ had a comparable effect on nasal symptoms, but LCZ was faster and displayed a greater effect on histamine wheal. PMID- 15345914 TI - Clinical significance of anticardiolipin and anti-beta2-glycoprotein I antibodies. AB - BACKGROUND: Anticardiolipin (aCl) and anti-beta2-glycoprotein I (anti-beta2-gpI) antibodies are autoantibodies associated with the antiphospholipid syndrome (APS), which is characterized by both arterial and venous thrombosis and miscarriages. The scope of this study was to explore the clinical characteristics of patients with aCl and anti-beta2-gpI antibodies. METHODS: ACl were tested in 3,600 consecutive sera in our laboratory between January 1999 and June 2001. The clinical diagnosis and prevalence of thrombosis and pregnancy morbidity were retrospectively reviewed in aCl-positive patients. Furthermore, the frequency of anti-beta2-gpI antibodies, lupus anticoagulant (LA), prolonged activated partial thromboplastin time (aPTT), and thrombocytopenia were investigated in aCl positive patients. RESULTS: 147 aCl-positive patients, 110 women and 37 men with a mean age of 41 years (range 7.8-82.5), were identified. 42 (28.6%) aCl-positive patients fulfilled the criteria for APS which was secondary to a connective tissue disorder in 8 patients. The frequency of anti-beta2-gpI antibodies and LA, prolonged aPTT, and thrombocytopenia in aCl-positive patients was 23.8, 27.2, 25.7 and 9.2%, respectively. The presence of both aCl and anti-beta2-gpI antibodies was strongly associated with clinical symptoms of APS (p = 0.007) compared to p = 0.008 for LA. CONCLUSION: Our data suggest that assessment of anti-beta2-gpI antibodies in addition to aCl is a valuable diagnostic tool in the workup of patients with APS. PMID- 15345915 TI - Canine mast cell activation via human IgG1 and IgG4. AB - BACKGROUND: We have reported that canine mastocytoma-derived CM-MC cells are activated via canine IgG and express a high-affinity IgG receptor (canine FcgammaRI). The predicted amino acid sequence of the canine FcgammaRI alpha subunit was found to be 72% similar to that of humans. These results suggest that canine FcgammaRI have binding activity with human IgG and led us to investigate CM-MC activation via canine FcgammaRI and human IgG. METHODS: The binding of human IgG to canine FcgammaRI was examined by flow cytometry using FITC conjugated human IgG. [Ca2+]i increase or histamine release via canine FcgammaRI and the four human IgG subclasses was measured following aggregation of IgG-bound FcgammaRIs by anti-human IgG. To determine the binding activity of canine FcgammaRI with human IgG1 or IgG3, the displacement of 125I-labeled canine IgG from canine FcgammaRI was examined by unlabeled human IgG1 or IgG3. RESULTS: The fluorescence intensity of CM-MC cells was markedly (about 50 times) elevated by incubation with FITC-human IgG compared with the fluorescence of the control cells. A significant (p < 0.01) calcium response and histamine release were observed following aggregation of canine FcgammaRIs bound with human IgG1 or IgG4. 125I-labeled canine IgG was displaced from canine FcgammaRI by preincubation with unlabeled total human IgG or human IgG1 dose-dependently, whereas no displacement was detected by preincubation with human IgG3. CONCLUSIONS: Canine FcgammaRI possesses a significant binding activity with human IgG1 or IgG4, while IgG2 or IgG3 did not significantly react with canine FcgammaRI on CM-MC cells. PMID- 15345916 TI - Serum levels of interleukin-18 are elevated in the subacute phase of kawasaki syndrome. AB - BACKGROUND: Elevations of various cytokines, including Th1 and Th2 cytokines, have been reported in the acute phase of Kawasaki syndrome (KS). As interleukin (IL)-18 plays an important role in the Th1 cell response, investigating the relevance of IL-18 in KS should be helpful in determining the pathophysiology of KS. Therefore, we examined the IL-18 values in KS. METHODS: Serum IL-18 values were measured by an enzyme-linked immunosorbent assay. Samples were obtained from 41 patients in the acute and subacute phase of KS, 35 age-matched febrile controls and 13 afebrile controls. RESULTS: No difference was observed in the values of white blood cell counts or C-reactive protein between acute-phase KS patients and febrile controls. On the contrary, acute-phase KS patients showed a significantly lower mean IL-18 value (398 +/- 206 pg/ml) than that of febrile controls (584 +/- 307 pg/ml) (p = 0.006). Subacute-phase KS patients showed a significantly elevated level of IL-18 (517 +/- 276 pg/ml) compared to acute-phase patients (p = 0.0008). The IL-18 values in the subacute-phase patients showed a significant positive correlation with the duration of fever (r = 0.427, p = 0.0055) and also with the presence of coronary artery abnormalities (r = 0.332, p = 0.0340). The incidence of elevated IL-18 values in the subacute-phase patients was significantly higher than that in the afebrile controls (p = 0.048). CONCLUSIONS: Patients with KS showed normal IL-18 values in the acute phase and elevated values in the subacute phase. IL-18 pathways were activated in the subacute phase of KS, and subacute IL-18 values might be reflected in the severity of KS. PMID- 15345917 TI - Genetic heterogeneity in autosomal dominant pseudohypoaldosteronism type I: exclusion of claudin-8 as a candidate gene. AB - BACKGROUND/AIMS: Pseudohypoaldosteronism type I (PHAI) is an inherited disorder characterized by renal salt wasting, hyperkalemic metabolic acidosis, and hyperaldosteronism. Its known causes are mutations in the mineralocorticoid receptor and the epithelial sodium channel (ENaC), but there are reports of genetic heterogeneity. Claudin-8 is a tight junction protein that acts as a paracellular cation barrier in the distal nephron. The aim of this study was to test the hypothesis that mutations in claudin-8, which would be expected to induce a distal tubule cation leak, can be a cause of PHAI. METHODS: We identified 10 patients with autosomal dominant PHAI in whom mutations in the mineralocorticoid receptor and ENaC had been excluded. The claudin-8 gene and upstream region was sequenced in all patients. RESULTS: No disease-associated claudin-8 mutations were identified. A novel polymorphic allele in the 3' untranslated region was identified in 2 patients, but was also found in 15% of individuals in a panel of normal controls. CONCLUSION: We present further evidence for locus heterogeneity in PHAI. Mutations in claudin-8 are unlikely to be a cause of PHAI. Further studies of other claudins in this disease are warranted. PMID- 15345929 TI - Using thoracic epidural catheters for pain management. PMID- 15345918 TI - Influence of the type of membrane and heparin on serum lipid parameters during a dialysis session: a pilot study. AB - BACKGROUND/AIM: The type of heparin and membrane used might influence the lipids in patients on hemodialysis (HD). However, there are limited and debatable data concerning the lipid changes during an HD session. The aim of our study was to examine the changes in serum lipid parameters during the HD session in relation to the heparin and dialysis membrane used. METHODS: Ten patients on HD 3 times/week participated in the study. The study was performed in three phases (A, B, C), each of 1 week's duration. The types of membranes used were Hemophan (phase A), ethylene vinyl alcohol (phase B) and polyacrylonitrile (phase C), respectively, in a random order. During the midweek session of each phase we used classic heparin, while during the session at the end of the week low molecular weight heparin was administered. Serum total cholesterol, triglycerides, HDL cholesterol, Lp(a), albumin and total proteins were measured before and 5 min after the HD and hourly during the HD session. RESULTS: In all phases, we found a progressive increase in all lipid parameters during the HD session, except Lp(a). This increase, however, was possibly due to hemoconcentration. CONCLUSIONS: This pilot study showed that (1) the type of heparin and membrane used does not seem to affect the serum lipid profile during a single HD procedure, and (2) the changes observed in serum lipid parameters are mainly due to hemoconcentration. PMID- 15345930 TI - Introducing patient-controlled oral analgesia. PMID- 15345931 TI - Burning beds. PMID- 15345932 TI - Documenting short-staffing: a delicate balance. PMID- 15345933 TI - What is "renal dose" dopamine? PMID- 15345948 TI - Abdominal trauma: dealing with the damage. PMID- 15345949 TI - Taking a stand against fear. PMID- 15345951 TI - Patient education series. Coping with grief. PMID- 15345950 TI - Nosing into flu season. AB - Learn how to administer influenza vaccine intranasally in a few steps. PMID- 15345955 TI - Locking down tetanus. PMID- 15345956 TI - A different kind of shower. PMID- 15345957 TI - More Americans than ever use CAM, says CDC. PMID- 15345960 TI - What you should know about needleless I.V. systems. PMID- 15345961 TI - Bloodborne pathogen exposure. PMID- 15345962 TI - A U.S. Army Forward Surgical Team's experience in Operation Iraqi Freedom. AB - BACKGROUND: The Forward Army Surgical Team (FST) was designed to provide surgical capability far forward on the battlefield to stabilize and resuscitate those soldiers with life and limb threatening injuries. Operation Iraqi Freedom represents the largest military operation in which the FST concept of health care delivery has been employed. The purpose of our review is to describe the experience of the 555FST during the assault phase of Operation Iraqi Freedom. METHODS: During the 23 days beginning 21 March 2003, data on all patients seen by the 555 FST were recorded. These data included combatant status, injuries according to anatomic location, and operative procedures performed. RESULTS: During the twenty-three day period, the 555 FST evaluated 154 patients. There were 52 EPWs, 79 U.S. soldiers, and 23 Iraqi civilians treated. Injuries to the lower extremity and chest (48% and 25%) were the most common in the EPW group. Upper extremity and lower extremity injuries were the most common in the civilian (57% and 39%) and U.S. soldier groups (32% and 30%). The number of injured regions per patient were 1.14 for U.S. soldiers, 1.33 for EPWs, and 1.52 for Iraqi civilians (p < 0.003). EPWs had proportionately more thoracic and abdominal injuries than the other groups (p < 0.05). CONCLUSIONS: Majority of the life threatening injuries evaluated involved EPWs. A combination of body armor and armored vehicles used by U.S. soldiers limited the number of torso injuries presenting to the FST. Early resuscitation and stabilization of U.S. soldiers, EPWs, and civilians can be successfully accomplished at the front lines by FSTs. Further modification of the FST's equipment will be needed to improve its ability in providing far forward surgical care. PMID- 15345963 TI - Fatal and non-fatal injuries among U.S. Air Force personnel resulting from the terrorist bombing of the Khobar Towers. AB - BACKGROUND: Culminating with the events of September 11, 2001, U.S. civilian and military populations are vulnerable to terrorist attacks. Information about the patterns of injuries and their causes inform disaster management planning and can improve structural and architectural design. This report documents the injuries following an estimated 20K bomb at the Khobar Towers military compound in Dhahran, Saudi Arabia. METHODS: US Air Force personnel in the compound at the time of the bombing were evaluated. Data were collected through a mail survey, and a review of military medical records and autopsy reports. Outcome measures included injury types, treatment status, severity, and causes of injuries. RESULTS: The study identified 574 persons who were injured in the bombing; 19 persons died. A total of 420 (73%) persons were injured directly in the blast and 154 (27%) persons were injured during evacuation, search and rescue, during cleanup, or sustained only auditory or inhalation trauma. Sixteen percent of survivors injured directly in the blast were hospitalized; 6% were critical. Soft tissue and foreign body injuries of the lower extremities were the most common injuries among survivors, most often caused by glass. Persons who died suffered extensive, multiple injuries from blunt trauma and one death was caused by glass. CONCLUSIONS: All deaths were immediate and the majority of survivors suffered minor to moderate injuries. Injuries and deaths were consistent with extensive glass damage in the compound and fragmentation of building concrete. Improved building designs and retrofits such as blast-resistant glass and prevention of structural collapse or building fragmentation should be considered in building design and construction in high threat areas. PMID- 15345964 TI - Effects of low temperature on shear-induced platelet aggregation and activation. AB - BACKGROUND: Hemorrhage is a major complication of trauma and often becomes more severe in hypothermic patients. Although it has been known that platelets are activated in the cold, studies have been focused on platelet behavior at 4 degrees C, which is far below temperatures encountered in hypothermic trauma patients. In contrast, how platelets function at temperatures that are commonly found in hypothermic trauma patients (32-37 degrees C) remains largely unknown, especially when they are exposed to significant changes in fluid shear stress that could occur in trauma patients due to hemorrhage, vascular dilation/constriction, and fluid resuscitation. METHODS: Using a cone-plate viscometer, we have examined platelet activation and aggregation in response to a wide range of fluid shear stresses at 24, 32, 35, and 37 degrees C. RESULTS: We found that shear-induced platelet aggregation was significantly increased at 24, 32, and 35 degrees C as compared with 37 degrees C and the enhancement was observed in whole blood and platelet-rich plasma. In contrast to observation made at 4 degrees C, the increased shear-induced platelet aggregation at these temperatures was associated with minimal platelet activation as determined by the P-selectin expression on platelet surface. Blood viscosity was also increased at low temperature and the changes in viscosity correlated with levels of plasma total protein and fibrinogen. CONCLUSION: We found that platelets are hyper reactive to fluid shear stress at temperatures of 24, 32, and 35 degrees C as compared with at 37 degrees C. The hyperreactivity results in heightened aggregation through a platelet-activation independent mechanism. The enhanced platelet aggregation parallels with increased whole blood viscosity at these temperatures, suggesting that enhanced mechanical cross-linking may be responsible for the enhanced platelet aggregation. PMID- 15345965 TI - Thermal injury resulting from application of a granular mineral hemostatic agent. AB - BACKGROUND: Uncontrolled hemorrhage accounts for the majority of deaths in combat. Effective topical hemostatic agents suitable for use on the battlefield may be valuable in controlling hemorrhage until definitive surgical intervention is possible. In an effort to identify a hemostatic agent suitable for battlefield use, we evaluated several potential hemostatic agents in a swine injury model and noted thermal injury to tissues with a granular mineral hemostatic agent (QuikClot). METHODS: Anesthetized swine were maintained with a mean arterial pressure in excess of 60 mm Hg. Cutaneous, muscular, hepatic, splenic, venous, and arterial wounds were created in a standardized fashion. Topical hemostatic agents were immediately applied to the wounds and the amount of bleeding and time to hemostasis were noted. RESULTS: The results reported here are part of a larger study in which a variety of hemostatic agents were evaluated. Only the findings related to the granular mineral hemostatic agent are discussed here. Application of the agent resulted in elevated tissue surface temperatures in excess of 95 degrees C and internal tissue temperatures exceeding 50 degrees C, 3 mm deep to the bleeding surface. Necrosis of fat and muscle were noted as well as full and partial thickness cutaneous burns. CONCLUSIONS: Topical administration of a granular mineral hemostatic agent to a variety of wounds in an experimental swine model resulted in thermal tissue injury and necrosis. Suggestions for reducing the extent of injury with this product are offered. PMID- 15345966 TI - Treatment of field water with sodium hypochlorite for surgical irrigation. AB - BACKGROUND: Early irrigation and surgical debridement of high-energy wounds and open fractures effectively prevents infection. Rapid wound care has been maximized by the United States military's "forward surgical teams." However, the volume of sterile irrigant required to treat multiple patients with multiple wounds presents a significant logistical burden. Using ground-derived field water could eliminate this burden. METHODS: We collected 100 water samples from five sources. An initial bacterial count (CFU/mL) was determined before treatment. 5% sodium hypochlorite was then added to each sample to derive a concentration of 0.025%. After treatment, a final bacterial colony count was performed. RESULTS: We found no bacterial growth in 99/100 samples. One post-treatment sample grew a single colony of a Bacillus species not present in the pretreatment culture and was determined to be an air contaminant. CONCLUSIONS: Our field-expedient modification of Dakin's solution could substitute for sterile irrigation fluid when it is neither available nor logistically feasible. PMID- 15345967 TI - Dynamic effects of a 9 mm missile on cadaveric skull protected by aramid, polyethylene or aluminum plate: an experimental study. AB - BACKGROUND: Most military helmets are designed to prevent penetration by small firearms using composite materials in their construction. However, the transient deformation of the composite helmet during a non penetrating impact may result in severe head injury. METHOD: Two experimental designs were undertaken to characterize the extend of injuries imparted by composite panels using in protective helmets. In the first series, 21 dry skulls were protected by polyethylene plates, with gaps between the protective plate and skull ranging from 12 to 15 mm. In another design, using 9 cadavers, heads were protected by aluminum, aramid, or polyethylene plates. Specimens were instrumented with pressure gauges to record the impact response. The ammunition used in these experiments was 9 mm caliber and had a velocity of 400 m/s. A macroscopic analysis of the specimens quantified fractures and injuries, which were then related to the measured pressures. RESULTS: Protective plates influenced both the levels of injury and the intracranial pressure. Injuries were accentuated as the plates was changed from aluminum to composite materials and ranged from skin laceration to extensive skull fractures and brain contusion. Fractures were associated with brain parenchymal pressures in excess of 560 kPa and cerebrospinal fluid pressure of 150 kPa. An air gap of a few millimeters between the plate and the head was sufficient to decrease these internal pressures by half, significantly reducing the level of injury. CONCLUSIONS: Ballistic helmets made of composite materials could be optimized to avoid extensive transient deformation and thus reduce the impact and blunt trauma to the head. However, this deformation cannot be completely removed, which is why the gap between the helmet and the head must be maintained at more than 12 mm. PMID- 15345968 TI - Laser Doppler flow and brain tissue PO2 after cortical impact injury complicated by secondary ischemia in rats treated with arginine. AB - BACKGROUND: Traumatic brain injury (TBI) makes the brain susceptible to secondary insults such as ischemia. This study tested the hypothesis that L-arginine would increase regional CBF (rCBF) and brain tissue PO2 (PbtO2) at the injury site. METHODS: A secondary insult model was employed in rodents. rCBF was measured with laser doppler flowmetry (LDF) and PbtO2 with a PO2 catheter at the impact site. Animals were randomized to receive L-arginine, D-arginine or saline intravenously, 5 minutes after impact. RESULTS: In animals who received L arginine, the percentage rCBF from baseline (%CBF) was higher at the impact site after impact (p < 0.001), during bilateral carotid occulation (BCO) (p = 0.001) and during reperfusion (p = 0.032). In contrast, PbtO2 was not significantly increased throughout the experiment for the L-arginine group. CONCLUSIONS: Administration of L-arginine increased rCBF in the injured brain tissue, and resulted in better preservation of CBF during BCO than D-arginine and saline. PMID- 15345969 TI - Early tracheostomy versus prolonged endotracheal intubation in severe head injury. AB - BACKGROUND: To see if early tracheostomy (fifth day) reduces duration of mechanical ventilation, ICU stay, incidence of pneumonia and mortality in comparison with prolonged intubation (PI) in patients with head injury. METHODS: Patients were prospectively included in this study if they met the following criteria: isolated head injury, Glasgow coma scale (GCS) score < or =8 on first and fifth day, with cerebral contusion on CT scan. On the fifth day, randomization was done in two groups: early tracheostomy group (T group, n = 31) and prolonged endotracheal intubation group (I group, n = 31). We evaluated total time of mechanical ventilation, ICU stay, pneumonia incidence and mortality. Complications related to each technique were noted. Analysis of data were performed using Yates and Kruskall Walis tests. p < 0.05 was considered significant. RESULTS: The two groups were comparable in term of age, sex, and Simplified Acute Physiologic Score (SAPS). The mean time of mechanical ventilatory support was shorter in T group (14.5 +/- 7.3) versus I group (17.5 +/ 10.6) (p = 0.02). After pneumonia was diagnosed, mechanical ventilatory time was 6 +/- 4.7 days for ET group versus 11.7 +/- 6.7 days for PEI group (p = 0.01). There was no difference in frequency of pneumonia or mortality between the two groups. CONCLUSION: In severe head injury early tracheostomy decreases total days of mechanical ventilation or mechanical ventilation time after development of pneumonia. PMID- 15345970 TI - Prognosis of traumatic head injury in South Tunisia: a multivariate analysis of 437 cases. AB - BACKGROUND: This study aimed to determine predictive factors of mortality after posttraumatic brain injury. METHODS: A retrospective study conducted over a 3 year period (1997-1999) involved 437 adult patients with head injury admitted to the intensive care unit of a university hospital in Sfax, Tunisia. Basic demographic, clinical, biologic, and radiologic data were recorded at admission and during the intensive care unit stay. RESULTS: This study included 393 men (90%) and 44 women with a mean age of 36 +/- 17 years. Traffic accidents were the main cause of trauma (85.6%). In 58% of the cases, the injury was serious (Glasgow Coma Score, <8). The mean simplified acute physiology score was 39 +/- 15, and the mean Injury Severity Score was 34.5 +/- 17. Of the 437 patients, 127 (29.1%) died. According to multivariate analysis, the factors that correlated with a poor prognosis were age older than 40 years (p < 0.01), simplified acute physiology score exceeding 40 (p < 0.001), Glasgow Coma Score lower than 7 (p = 0.03), intracranial mass lesion (p = 0.02), a cerebral herniation (p < 0.001), diabetes insipidus (p < 0.001), and blood sugar level higher than 10 mmol/L (p < 0. 001). CONCLUSIONS: In Tunisia, head injury is a frequent cause of hospitalization, comprising 14.4% of all adult admissions. It is observed most often among young patients involved in traffic accidents. The short-term prognosis is poor, with a high (29%) mortality rate, and determined by demographic, clinical, radiologic, and biologic factors. Prevention is highly advised. PMID- 15345971 TI - D-lactate increases pulmonary apoptosis by restricting phosphorylation of bad and eNOS in a rat model of hemorrhagic shock. AB - Resuscitation with racemic lactated Ringer's solution (containing equal amounts of D and L isomers of lactate) has been shown to induce pulmonary apoptosis. Substitution of DL-isomer lactate with ketone bodies (beta-hydroxybutyrate, BHB), sodium pyruvate, or L-isomer of lactate decrease this injury without changing the energy status of the tissues or the expression of apoptotic genes. These modified solutions however alter the function of apoptotic proteins through an unknown mechanism. We postulated that DL-LR induces apoptosis by restricting the phosphorylation of key apoptotic proteins. METHODS: Male Sprague Dawley rats (n = 30, 5/group) were subjected to a three stage, volume-controlled hemorrhage and randomized to the following groups. 1) No hemorrhage (Sham); 2) Hemorrhage and no resuscitation (NR); 3) Resuscitation with 3x shed blood volume of racemic LR (DL LR); 4) Resuscitation with 3x shed blood volume of LR containing only the L isomer of lactate (L-LR); 5) Resuscitation with 3s shed blood volume of pyruvate Ringer's (PR); 6) Resuscitation with 3s shed blood volume of ketone Ringer's (KR). The modified Ringer's solutions were identical to racemic LR except for equimolar substitution of DL-lactate for L-lactate, pyruvate and BHB respectively. Lung tissue was obtained 2 hours later and subjected to Western Blotting. The levels of Akt, Bad, and eNOS (total and phosphorylated) proteins were measured. Finally, the expression of gene coding for protein 14-3-3 was measured using RT-PCR. RESULTS: Resuscitation with DL-LR caused a significant (p < 0.05) increase in the total Bad and a decrease in phosphorylated Bad protein expression in the lung. It also caused an increase in the phosphorylated Akt levels and a decrease in gene coding for protein 14-3-3. These changes were consistent with signaling imbalances that favor apoptosis. Modified LR solutions, on the other hand, did not cause these alterations. Phosphorylation pattern of eNOS supported the involvement of PI3K/Akt pathway in this process. CONCLUSION: Racemic lactate plays a role in the induction of pulmonary apoptosis by restricting phosphorylation of Bad and eNOS proteins. PMID- 15345972 TI - The usefulness of transcatheter arterial embolization for patients with blunt polytrauma showing transient response to fluid resuscitation. AB - BACKGROUND: This study aimed to determine whether nonsurgical management using transcatheter arterial embolization (TAE) is safe for patients with blunt multiple trauma who transiently respond to the initial fluid resuscitation. METHODS: Contrast computed tomography was performed for patients with blunt abdominal injuries, excluding those who did not respond to initial fluid resuscitation. Angiography was performed for patients with injuries showing contrast extravasation or solid organ injury classified, according to the American Association for the Surgery of Trauma, as grade 3 or higher on computed tomography. Transcatheter arterial embolization was performed when angiography showed arterial extravasation. The protocol was abandoned for any patients who became profoundly hypotensive (with systolic blood pressure 60 mm Hg or lower) during computed tomography or angiography. RESULTS: Between January 2000 and December 2002, 269 patients with blunt abdominal injuries underwent TAE immediately after admission. Of these patients, 41 had injuries in at least two regions and underwent TAE for these regions. Among them, 22 patients were hemodynamically stable or showed rapid response to fluid resuscitation. The nonsurgical treatment was successful in all these cases. The remaining 19 patients (Injury Severity Score, 37.3 +/- 8.2), who showed a transient response, were the subjects of this study. Of these patients, 15 underwent TAE for injuries in two regions (13 pelvic fractures, 7 splenic injuries, 6 hepatic injuries, 3 facial bleeding, and 1 renal injury), and 4 patients underwent TAE for injuries in three regions (4 had splenic injuries, 3 hepatic injuries, 2 renal injuries, 2 pelvic fractures, and 1 facial bleeding). For all these patients, TAE was successfully performed. Before TAE, the systolic blood pressure was 79.9 +/- 8.4 mm Hg, and the shock index was 1.45 +/- 0.25 mm Hg. After TAE, the corresponding values were 120.6 +/- 19.3 mm Hg and 0.87 +/- 0.16 mm Hg, respectively (p < 0.001). The rate of fluid administration required after TAE (214.2 +/- 139.3 mL/hour) was significantly less than that required before TAE (1244.2 +/- 347.1 mL/hour; range, 632-1,728 mL/hour) (p < 0.001). The deaths of two patients were classified as nonpreventable on the basis of the Trauma and Injury Severity Score (TRISS), and their respective probabilities of survival were determined to be 0.13 and 0.03. CONCLUSION: Nonsurgical management using TAE can be performed safely even for patients with blunt multiple trauma who are in hemorrhagic hypotension if their hemodynamics are improved by resuscitation with 2 L of fluid. PMID- 15345973 TI - Free abdominal fluid on ultrasound in unstable pelvic ring fracture: is laparotomy always necessary? AB - BACKGROUND: In unstable pelvic ring fractures free abdominal fluid on ultrasound (US) may be caused by retroperitoneal hematoma that passes into the abdominal cavity or by an additional intraabdominal lesion. In this study a clinical pathway for the therapy of potentially combined lesions was analyzed. PATIENTS AND METHODS: All patients treated in the ED for severe trauma underwent basic sonographical and radiologic diagnostics within 15 minutes. of admission. Data were prospectively documented. According to the treatment protocol unstable pelvic ring fractures with initial free fluid on US received laparotomy. Patients with stable vital conditions had abdominal CT-Scan before surgery. RESULTS: 1472 consecutive severely injured patients (ISS 20, age: 39 years) were included. Eighty subjects had sustained type B (47) or C (33) pelvic ring fracture. Early free abdominal fluid on US was absent in 49 cases. Three patients in this group required celiotomy later on, during ICU treatment. In 31 patients free fluid was present. All of them had laparotomy. Only one patient showed retroperitoneal hematoma alone, while all others had one or more significant lesions (rupture) that required surgical repair. Simultaneously with laparotomy pelvic stabilization was performed by external (19) or internal (6) fixation. In all cases with massive pelvic hemorrhage and free fluid in US bleeding was controlled by internal tamponade and external fixation. CONCLUSION: The finding of intraperitoneal fluid on US in the emergency department strongly correlates with significant intraabdominal lesions requiring surgical intervention. Early laparotomy appears indicated in these cases. Shock control in pelvic bleeding can be sufficiently achieved by internal tamponade and external fixation. PMID- 15345974 TI - Hand-held thoracic sonography for detecting post-traumatic pneumothoraces: the Extended Focused Assessment with Sonography for Trauma (EFAST). AB - BACKGROUND: Thoracic ultrasound (EFAST) has shown promise in inferring the presence of post-traumatic pneumothoraces (PTXs) and may have a particular value in identifying occult pneumothoraces (OPTXs) missed by the AP supine chest radiograph (CXR). However, the diagnostic utility of hand-held US has not been previously evaluated in this role. METHODS: Thoracic US examinations were performed during the initial resuscitation of injured patients at a provincial trauma referral center. A high frequency linear transducer and a 2.4 kg US attached to a video-recorder were used. Real-time EFAST examinations for PTXs were blindly compared with the subsequent results of CXRs, a composite standard (CXR, chest and abdominal CT scans, clinical course, and invasive interventions), and a CT gold standard (CT only). Charts were reviewed for in-hospital outcomes and follow-up. RESULTS: There were 225 eligible patients (207 blunt, 18 penetrating); 17 were excluded from the US examination because of battery failure or a lost probe. Sixty-five (65) PTXs were detected in 52 patients (22% of patients), 41 (63%) being occult to CXR in 33 patients (14.2% whole population, 24.6% of those with a CT). The US and CXR agreed in 186 (89.4%) of patients, EFAST was better in 16 (7.7%), and CXR better in 6 (2.9%). Compared with the composite standard, the sensitivity of EFAST was 58.9% with a likelihood ratio of a positive test (LR+) of 69.7 and a specificity of 99.1%. Comparing EFAST directly to CXR, by looking at each of 266 lung fields with the benefit of the CT gold standard, the EFAST showed higher sensitivity over CXR (48.8% versus 20.9%). Both exams had a very high specificity (99.6% and 98.7%), and very predictive LR+ (46.7 and 36.3). CONCLUSION: EFAST has comparable specificity to CXR but is more sensitive for the detection of OPTXs after trauma. Positive EFAST findings should be addressed either clinically or with CT depending on hemodynamic stability. CT should be used if detection of all PTXs is desired. PMID- 15345975 TI - Prospective study of the clinical predictors of a positive abdominal computed tomography in blunt trauma patients. AB - BACKGROUND: CT scans are often used in the evaluation of blunt trauma patients. Many scans are negative. Clinical predictors of positive abdominal CT scans would be beneficial in patient care. METHODS: A prospective study of 213 patients at a Level I trauma center presenting with blunt trauma who underwent abdominal CT scan. Indications for CT scan were analyzed statistically, using univariate and multivariate models. RESULTS: Univariate chi2 tests showed abnormal pelvis x-ray (p = 0.0002) and an intubated patient (p = 0.03) were predictors of a positive CT scan. When subjected to multivariate logistic regression, these two indications were significant predictors of a positive CT scan, abnormal pelvis x-ray (p = 0.0005, OR=6.6, 95% CI), and an intubated patient (p = 0.02, OR=2.6, 95% CI). Univariate chi2 tests also showed that alcohol intoxication was statistically significant predictor of a negative CT scan (p = 0.03). CONCLUSION: Our data suggest that an abnormal pelvis x-ray and intubation are significant risk factors for a positive CT scan. Alcohol intoxication, mechanism of injury, and unreliable examination, without other associated indication for a scan, may warrant further study. PMID- 15345976 TI - Blunt cardiac trauma caused by fatal falls from height: an autopsy-based assessment of the injury pattern. AB - BACKGROUND: Falls from height are contributing widely to population morbidity and mortality, especially in urban settings. The presence of blunt cardiac injuries can increase morbidity among these patients, leading even to death. Some clinical studies and case reports have been published on the subject, but a systematic autopsy-based approach to the subject is missing in the literature of recent decades. METHODS: This study reviewed 61 cases of fatal fall from height that were subjected to a full autopsy at the Institute of Legal Medicine, Hamburg, Germany, from 1998 to 2002. The autopsy protocols and available clinical information were evaluated for assessment of the cardiac injury pattern. RESULTS: Cardiac injuries were found in 33 cases (54%), all of which involved falls from heights exceeding 6 m. In 16 cases, the cardiac injuries were the cause of death or contributed to the fatal outcome. In five of these cases, the individuals possibly could have recovered from their trauma if their heart injury had been sufficiently diagnosed and adequately treated in time. The most frequent finding was pericardial tearing (45%). Tears caused by stretching of the epicardium in the area wherein the inferior vena cava leads into the right atrium and epicardial hematoma were present in 11 cases (33%). Endocardial tears of the atria were found in six cases (18%), and did not occur during falls from heights lower than 11 m. Transmural tears to the right atrium were present in 10 cases (39%), and to the left atrium in 6 cases (18%). These tears occurred with increasing frequency in relation to greater heights. When the heights were lower than 15 m, these tears were smaller than 1 cm in diameter, but when the heights exceeded 15 m, extensive irregular tears were observed. Sternal fractures were seen in 76% of all cases involving heart injuries. In 16% of these cases, the fractures were multiple. Only 18% of the cases without cardiac injuries had sternal fractures, and none of these was multiple. Thus, the presence of severe sternal fractures can be used as an indicator of possible cardiac trauma. CONCLUSIONS: A thorough cardiologic diagnosis should always be performed for patients who survive a fall from height. These patients should be transported to a unit capable of performing cardiopulmonary bypass, and explorative thoracotomy should be considered. PMID- 15345977 TI - Selective operative management of major blunt renal trauma. AB - BACKGROUND: We reviewed the management and outcomes of patients at our Level I trauma center suffering major blunt renal trauma diagnosed and staged by CT scan. METHODS: We retrospectively reviewed the cases of 26 patients with blunt trauma at our institution who were initially hemodynamically stable and diagnosed with grade 4 or 5 renal injuries by CT scan. Patients were broken down into two groups based on whether they were managed conservatively or surgically. Patient characteristics and morbidity were analyzed. RESULTS: There were 14 patients managed conservatively and 12 patients managed surgically. There was no statistically significant difference in morbidity between the two groups. The only statistically significant predictor of failure of conservative management was a coexisting solid organ intra-abdominal injury. CONCLUSIONS: Conservative management of major blunt renal trauma is appropriate in hemodynamically stable patients. PMID- 15345978 TI - Renovascular injury: an argument for renal preservation. AB - BACKGROUND: Renovascular injury is uncommon among children. This study hypothesized that preservation of the severely injured kidney can be achieved safely without renal insufficiency, postinjury hypertension, or the need for hemodialysis. METHODS: Retrospective chart review of renal injuries seen between 1997 and 2001 at a level 1 pediatric trauma center was conducted. Severity of injury was graded by the American Association for the Surgery of Trauma Organ Injury Severity Scale. The outcome variables included the need for hemodialysis, impaired renal function (creatinine), and postinjury hypertension. RESULTS: In this study, 34 children presented with grade 1, 2, or 3 injury (74%), whereas 13 children presented with grade 4 or 5 renovascular injury (28%). The children with unilateral renovascular injury who underwent either nephrectomy or renal preservation had comparable outcomes with no hypertension, hemodialysis, or renal insufficiency in either group. CONCLUSIONS: The treatment outcomes were not different between the patients who underwent renal preservation and those who had immediate nephrectomy. The authors conclude that renal preservation should be attempted for all children with grade 4 or 5 renovascular injury. PMID- 15345979 TI - Is bilateral protected specimen brush sampling necessary for the accurate diagnosis of ventilator-associated pneumonia? AB - BACKGROUND: Clinical acumen alone is unreliable in establishing a diagnosis of ventilator-associated pneumonia (VAP) and controversy exists over which diagnostic tools should be utilized to confirm a clinical suspicion of VAP. The purpose of this study was to determine the reliability of blind protected specimen brush (PSB) sampling in the diagnosis of VAP and if bilateral PSB sampling is necessary. METHODS: Prospective study comparing blind PSB sampling with bronchoscopic directed PSB sampling in thirty-four consecutive SICU patients with a clinical suspicion of VAP. All patients underwent blind PSB sampling followed by bronchoscopic directed contralateral PSB sampling. RESULTS: Twenty four of 34 patients (71%) were diagnosed to have VAP. The concordance rate between blind and directed PSB samples was 53% (18/34). When blind PSB was positive (15/34), the contralateral sample yielded a different microorganism in three patients (9%). When blind PSB was negative (19/34), infection was present in the contralateral lung in nine patients (26%). Blind PSB sampling alone was inaccurate in 35% of patients. CONCLUSIONS: The low concordance between blind and directed PSB suggests the need to sample both lung fields. Bilateral PSB sampling can identify unsuspected pathogenic microorganisms in the contralateral lung. PMID- 15345980 TI - Restraint use and lower extremity fractures in frontal motor vehicle collisions. AB - BACKGROUND: Seat belts and air bags have been shown to significantly reduce morbidity and mortality following MVCs. Research suggests that restraint use does not protect against lower extremity fracture; however, no population-based studies of this association exist. The purpose of this study is to compare the effectiveness of combined seat belt and airbag restraint systems with airbag alone, seat belt alone, and no restraints with respect to incidence and location of lower extremity fractures. METHODS: A retrospective analysis of front seat occupants involved in police-reported, tow-away frontal MVCs was conducted using data from the 1995 through 2000 National Automotive Sampling System (NASS). Incidence and relative risk (RR) of fracture to specific bony regions were measured according to seat belt use and airbag deployment. RESULTS: Compared with unrestrained occupants, occupants restrained with airbag only had significantly higher risk for all types of lower extremity fractures whereas those occupants restrained with either seat belt only or seat belt and airbag had lower risk of fracture. The greatest difference was seen with tibia/fibula fractures in airbag only (RR, 2.14) but this trend continued to be significant with femur and pelvic fractures (RR, 1.13 and 1.23, respectively). CONCLUSION: While airbags may reduce the risk of death when used alone or in combination with seat belts, the results of this study demonstrate that air bags increase the risk of lower extremity fractures when used as the sole method of passenger protection. Also, they may do so differentially according to skeletal region. This data strongly support the consideration of developing accessory knee bolster airbags to prevent the "submarining" or sliding under the airbag that may be responsible for this finding. PMID- 15345981 TI - Goal-directed ultrasound in the detection of long-bone fractures. AB - BACKGROUND: New portable ultrasound (US) systems are capable of detecting fractures in the remote setting. However, the accuracy of ultrasound by physicians with minimal ultrasound training is unknown. METHODS: After one hour of standardized training, physicians with minimal US experience clinically evaluated patients presenting with pain and trauma to the upper arm or leg. The investigators then performed a long-bone US evaluation, recording their impression of fracture presence or absence. Results of the examination were compared with routine plain or computer aided radiography (CT). RESULTS: 58 patients were examined. The sensitivity and specificity of US were 92.9% and 83.3%, and of the physical examination were 78.6% and 90.0%, respectively. US provided improved sensitivity with less specificity compared with physical examination in the detection of fractures in long bones. CONCLUSION: Ultrasound scans by minimally trained clinicians may be used to rule out a long-bone fracture in patients with a medium to low probability of fracture. PMID- 15345982 TI - Pressurization of the metastatic femur during prophylactic intramedullary nail fixation. AB - BACKGROUND: Venting during prophylactic nailing of metastatic femoral lesions may decrease intramedullary pressures and the risk of fat and tumor embolization. This study aimed to quantify maximum intramedullary pressure levels and durations, and to examine tumor displacement during prophylactic reamed intramedullary nailing of vented and nonvented metastatically involved femora. METHODS: Proximal metastatic defects were simulated in six pairs of cadaveric femora and prophylactically treated using reamed intramedullary nailing with or without proximal and distal vents. Maximum pressures, durations of pressure exceeding embolic threshold levels (25 and 200 mm Hg), and tumor displacement were recorded. RESULTS: Maximum pressures and durations of pressure exceeding embolic thresholds decreased more than 50% with venting (p < 0.05). Distal pressures surpassed the 25-mm Hg threshold even with venting. Post-test sectioning showed increased tumor disbursement in vented specimens. CONCLUSIONS: Venting reduces pressurization during prophylactic reamed intramedullary nailing of femoral metastases, but may increase the spread of tumor to extraskeletal tissue if vented tissue is not contained. PMID- 15345983 TI - Treatment of complex tibial plateau fractures using the less invasive stabilization system plate: clinical experience and a laboratory comparison with double plating. AB - BACKGROUND: Bicondylar tibial plateau fractures are complex injuries, historically associated with high complication rates. The purpose of this study was: 1) to evaluate the clinical use L.I.S.S plating system for stabilization of bicondylar tibial plateau fractures. 2) To compare the biomechanics of this plating system with a double plate construct. METHODS AND MATERIALS: Thirty-eight patients who sustained a complex tibial plateau fracture (OTA type 41C) at one of three level-one trauma centers were stabilized using the Less Invasive Stabilization System (L.I.S.S.). The cohort of patients was evaluated clinically and radiographically for outcomes at a mean 15 months. In phase 2 of this study a model of a bicondylar tibial plateau fractures was made in six matched pairs of embalmed, human tibia and randomized to fixation with either a L.I.S.S plate or a standard double plate construct. The tibias were then subjected to an axial cyclic load of 500N for 10 cycles (3Hz) to approximate 2 months in vivo and displacements measured. RESULTS: Thirty-six of /38 (95%) patients united at 4 months after surgery with no loss of fixation nor infection. Two patients underwent prophylactic autogenous bone grafting for bone loss and united by 3 months postgrafting. Significant loss of knee range of motion (<90) was seen in five patients.Biomechanically, no differences in permanent inferior displacement of the medial fragment were found in initial axial loading and after 10 cycles between the two plate constructs. However, when loaded to 500N the L.I.S.S plate construct demonstrated almost twice the displacement of the medial fragment compared with the dual plate construct. No specimen lost fixation during cycling. CONCLUSION: The L.I.S.S plating system provides stable fixation of complex bicondylar tibial plateau fractures allowing early range of knee motion with favorable clinical results. PMID- 15345984 TI - Chemiluminescence: comparison of whole blood with isolated polymorphonuclear leukocytes after major trauma. AB - INTRODUCTION: Neutrophil (PMN) mediated tissue injury is central to the development of post-traumatic ARDS/MOF. Changes in activity caused by PMN isolation may be avoided by studying respiratory burst activity using whole blood chemiluminescence (WBCL). METHODS: WBCL and PMNCL were measured in 5 normal laboratory volunteers (NLV) and 9 patients sustaining major trauma, within 2 hours of admission. Receptor mediated (STZ) and independent (PMA) activating agents were used. RESULTS: PMA activation confirmed significant priming both in WBCL and PMNCL after major trauma. With STZ, priming was confirmed in the WBCL study, but the trauma patient PMNCL showed no difference in response to those of NLV. Although the study population was small, those patients later developing ARDS demonstrated significantly greater STZ activated WBCL, 8 hours after admission. CONCLUSION: PMN isolation alters behavior in vitro. This may lead to important differences of in vivo PMN function being obscured when studied in the laboratory setting. Further study of CL response and surface receptor expression is clearly warranted, both in WB and PMN preparations. PMID- 15345985 TI - Violence-related injury and intimate partner violence in an urban emergency department. AB - BACKGROUND: To facilitate the identification of ED patients at risk for intimate partner violence (IPV), we assessed the relationship of acute violence-related injury and history of IPV victimization or perpetration. METHODS: : This cross sectional study systematically sampled patients presenting to an urban ED. Reason for visit, past year history of IPV victimization and perpetration, alcohol and drug use and abuse, and sociodemographic factors were assessed. We hypothesized that violence-related injury would be positively associated with a history of IPV victimization and with IPV perpetration. RESULTS: The odds of violence-related injury was increased three-fold among persons with a history of IPV victimization and nearly two-fold (although not statistically significant) among those with IPV perpetration history. Male gender, younger age, and problem drinking were independent risk factors in both models. CONCLUSION: Screening for IPV among individuals presenting with a violence-related injury may be helpful in identifying individuals at risk of partner violence. PMID- 15345986 TI - Posttraumatic distress, alcohol disorders, and recurrent trauma across level 1 trauma centers. AB - BACKGROUND: Injured survivors of individual and mass trauma receive their initial evaluation in acute care. Few investigations have comprehensively screened for posttraumatic stress disorder (PTSD) symptoms and related comorbidities across sites. METHODS: This investigation included 269 randomly selected injury survivors hospitalized at two level 1 trauma centers. All patients were screened for PTSD, depressive, and peritraumatic dissociative symptoms during their surgical inpatient admission. Prior traumatic life events and alcohol abuse/dependence also were assessed. RESULTS: In this study, 58% of the patients demonstrated high levels of immediate posttraumatic distress or alcohol abuse/dependence. Regression analyses identified greater prior trauma, female gender, nonwhite ethnicity, and site as significant independent predictors for high levels of posttraumatic distress. CONCLUSIONS: High levels of posttraumatic distress, recurrent trauma, and alcohol abuse/dependence were present in more than half of acute care inpatients. Early mental health screening and intervention procedures that target both PTSD and alcohol use should be developed for acute care settings. PMID- 15345987 TI - Prediction of psychological health after an accidental burn. AB - BACKGROUND: Burn victims often display psychological symptoms that can impede recovery, but knowledge about risk factors for psychopathology is limited. This study aimed to predict psychological health 3 months after burn injury from coping and trauma-related factors assessed early in hospitalization. METHODS: For this study, 34 burn patients were interviewed during hospitalization about their accident and coping. Questionnaires were administered during hospitalization and 3 months after the burn including the Impact of Event Scale-Revised for posttraumatic stress symptoms (intrusion, avoidance, arousal) and the Hospital Anxiety and Depression Scale for mood. RESULTS: Anxiety, depressive, and avoidant symptoms at 3 months were highly predicted by baseline levels of these symptoms and avoidant coping. The life threat at the burn event predicted intrusive and arousal symptoms, and coping by self-control predicted less intrusive symptoms. Burn severity was not predictive of psychological health. CONCLUSIONS: Coping style, life threat during the accident, and early symptoms are strong predictors of psychopathology after a burn. PMID- 15345988 TI - Improving risk adjustment in critically ill trauma patients: the TRISS-SAPS Score. AB - OBJECTIVES: To test (a) the prognostic performance of TRISS and SAPS II scoring systems in a large sample of trauma patients admitted to Austrian ICUs, and (b) the hypothesis that the prognostic performance of TRISS could be improved by adding SAPS II. METHODS: Prospective multicenter cohort study comprising 5,538 trauma patients out of 35,637 patients admitted to 31 ICUs in Austria over a 4 year period. RESULTS: Separately, TRISS and SAPS II showed lack of calibration in the cohort of trauma patients. The database was then split into two equal samples, development and validation. Using the development sample, a new scoring system was developed, with vital status at hospital discharge as the dependent variable and TRISS and SAPS II as independent variables. The prognostic performance of the new TRISS-SAPS system was then assessed in the validation cohort: Both, discrimination (as shown by area under the ROC curve), and calibration (using Hosmer-Lemeshow goodness-of-fit statistics) was excellent. CONCLUSIONS: We improved risk adjustment in critically ill trauma patients by combining TRISS and SAPS II. This new scoring system might aid in evaluating and comparing specialized trauma ICUs. PMID- 15345989 TI - Advanced trauma life support study: quality of diagnostic and therapeutic procedures. AB - BACKGROUND: The introduction of the ATLS course in The Netherlands in 1995 provided for an opportunity to compare data of trauma patients between a pre-ATLS and a post-ATLS period. MATERIALS AND METHODS: Over a 3-year period (May 1996 - September 1997 pre ATLS; December 1997-April 1999 post ATLS) 63 trauma patients with an AIS-ISS > or = 16 (n = 31, pre-ATLS and n = 32, post-ATLS) were prospectively studied in two community residency training (ACS Level III) hospitals. All diagnostic and therapeutic procedures were recorded by a video camera and evaluated by a neutral faculty of six experienced ATLS trained specialists. RESULTS: Ten out of 14 interventions were performed qualitatively better in the post-ATLS group, while also the overall score was highly significantly better (4.2 pre-ATLS and 5.8 post-ATLS, p < 0.0001). CONCLUSION: Using the opinion of an expert team, this study identified a significantly lower number of patients with inadequate management. PMID- 15345990 TI - Safe and delayed repair in acute aortic traumatic insufficiency based on echocardiographic criteria. PMID- 15345991 TI - Extracorporeal life support in pulmonary failure after traumatic rupture of the thoracic aorta: a case report. PMID- 15345992 TI - Chronic atlantoaxial rotatory fixation with anterolisthesis: case report. PMID- 15345993 TI - An orbitocranial knife injury without functional deficit in a child. PMID- 15345994 TI - Traumatic aneurysm of the superficial temporal artery: case report. PMID- 15345995 TI - Trauma to the long thoracic nerve and associated scapula winging in a low velocity rear-end automobile collision: case report. PMID- 15345996 TI - Splenic artery embolization for post-traumatic splenic artery pseudoaneurysm in children. AB - Although rare, traumatic splenic artery pseudoaneurysm (SAP) can be life threatening. The diagnostic approaches as well as the methods of treatment of SAP are yet to be determined. We present the case of a 10-year-old boy treated conservatively for a grade III blunt splenic injury (BSI). The child was discharged to home after a 5-day uneventful hospitalization but was found on routine follow-up CT scan to have a large SAP. The pseudoaneurysm was successfully angiographically embolized and subsequent abdominal CT demonstrated successful resolution of the pseudoaneurysm with a small residual splenic cyst. We reviewed the eight cases of post-traumatic SAP in children that have been published in the English literature. Unlike SAP in adult patients, the severity of the splenic injury does not have predictive value for development of SAP in children. Abdominal pain was the most frequent symptom of SAP, but three children were asymptomatic at the time of diagnosis. Therefore, the possibility of SAP should be investigated even in the asymptomatic child with mild splenic injury. When a splenic pseudoaneurysm is diagnosed, we believe splenic artery embolization is indicated. PMID- 15345997 TI - Blunt traumatic injury of the gallbladder. PMID- 15345998 TI - Working Group on Trauma Research Program summary report: National Heart Lung Blood Institute (NHLBI), National Institute of General Medical Sciences (NIGMS), and National Institute of Neurological Disorders and Stroke (NINDS) of the National Institutes of Health (NIH), and the Department of Defense (DOD). PMID- 15345999 TI - Massive bilateral pulmonary embolus requiring pulmonary embolectomy in a low-risk trauma patient. PMID- 15346000 TI - Injuries are the leading cause of death among children less than 15 years old. PMID- 15346003 TI - Let us set the records straight for the future of the communication systems. PMID- 15346004 TI - Congenital trismus secondary to masseteric fibrous bands: a 7-year follow-up report as an approach to management. AB - A 7-year prospective follow-up report, which was previously presented in this journal as an initial pediatric case report, is presented as an approach to management of congenital trismus secondary to masseteric fibrous bands. Adams and Rees discussed management, including endoscopic exploration at 18 months of age with early recurrence of trismus. Under the care of the same plastic surgeon and his team, the progress of this patient over 7 years has given us an insight into management. The cause of trismus is not fully elucidated, but the condition can result in compromised caloric intake, speech development, facial appearance, dental care, and oral hygiene. The decreased oral opening may be secondary to shortening of the muscles of mastication, which may cause tension moulding and distortion of the coronoid process; yet, there is no consensus on the optimal management of temporomandibular joint trismus and all its causes. The patient presented in this report, now aged 7 years, has proceeded through to open surgery on two occasions yet, regrettably, has persistently tight masseter muscles and only 8 mm of jaw opening. PMID- 15346005 TI - The role of bone morphogenetic proteins BMP-2 and BMP-4 and their related postreceptor signaling system (Smads) in distraction osteogenesis of the mandible. AB - Distraction osteogenesis has become a widely used clinical approach in the treatment of craniofacial and orthopedic disorders. The exact biological mechanism of bone formation remains illusive, however. The aim of this study was to evaluate the role of bone morphogenetic protein-2, bone morphogenetic protein 4, and transforming growth factor-beta superfamily-related postreceptor signaling glycoproteins Smads 1 through 5 in distraction osteogenesis. Twelve sheep randomly divided into two groups were distracted to 24 mm at 1 or 4 mm/d using a submandibular osteotomy and an external distractor. After a 5-week fixation period, the mandibles were harvested. Employing immunohistochemical techniques, the sections were investigated for the previous antigens. Osteoblasts and periosteal lining cells were strongly positive. The matrix did not stain for the antigens investigated. Osteocytes demonstrated weak staining for the antigens. No significant difference between the groups was noted. In fracture healing, bone morphogenetic proteins 2 and 4 have been localized to the cambial layer of the periosteum, where healing occurs by intramembranous ossification. Their diffuse staining of the osteoblasts in the distracted region supports a similar role in distraction osteogenesis, where bone formation is predominantly through intramembranous ossification. Furthermore, bone morphogenetic proteins 2 and 4 have been demonstrated to promote mesenchymal cell conversion to osteoblasts. This is similar to the process observed in distraction osteogenesis. The presence of related Smads confirms postreceptor activity of these bone morphogenetic proteins in the process of distraction osteogenesis. This study supports induction of bone morphogenetic proteins 2 and 4, their related postreceptor signaling system (Smads), and intramembranous bone formation associated with mechanical strain in distraction osteogenesis. PMID- 15346006 TI - Craniofacial reconstruction with high-density porous polyethylene implants. AB - Deformities of the facial skeleton may be reconstructed using autogenic or allogenic materials. Porous polyethylene is one of the few alloplastic materials currently in use having a well-documented history of reconstruction or augmentation in the maxillofacial region. High-density porous polyethylene, which is shown to be effective as a biomaterial, has additional advantages like tissue ingrowth, no capsule formation around it, and easy fixation. In this study, 83 implants in 71 patients were evaluated. Seven patients were in need of a second intervention. Three of the seven secondary interventions were for contour alignment, and four interventions were for extraction of the implants because of extrusion or infection. Placement of porous polyethylene implants directly under the skin without coverage of periosteum or another fascial envelope has an increased risk of early and especially late exposure. In cases like nasal dorsum or microtia reconstruction, we prefer autogenic grafts instead of allogenic materials. PMID- 15346007 TI - Rapid lengthening of rabbit mandibular ramus by using nitinol spring: a preliminary study. AB - Nitinol is well known for its shape memory and superelasticity, which means it can return to its original shape from large deflections and produce moderate forces in the process. Based on these characteristics, the authors attempted to use an internal Nitinol spring for automatic lengthening of mandibular ramus. Twenty adult white rabbits underwent transverse osteotomy in unilateral mandibular ramus to remove a 15.0 mm condylar segment and L-osteotomy in remnant ramus to create a vascularized transport disc. The S-shaped Nitinol spring, with two ends bent as perpendicular underprops, was constricted and secured across the transverse gap of L osteotomy by the underprops. Six animals were killed 6 days after operation for observation of bone transport rate. Other surviving rabbits were killed 8 weeks later for morphologic and histologic examination. Mechanical testing revealed moderate output of tensile elasticity by constricted spring on shape recovery in 37 degrees C. Rapid lengthening of mandibular ramus, more than 9.0 mm in 5 to 6 days, and new bone formation in 8 weeks were found. The radiograph and histologic appearance of regenerated bone was similar to those seen with traditional distraction osteogenesis. This preliminary study demonstrates the possibility of spring-mediated rapid mandibular lengthening across an osteotomy. However, the method needs additional scientific investigation for its clinical feasibility. PMID- 15346008 TI - The existence of the vomeronasal organ in human beings. AB - The frequency, location, and function of the vomeronasal organ, also known as the Jacopson organ, in human beings remains poorly understood. In this study, a search for the frequency of the vomeronasal organ was performed by nasal examinations of 346 adult patients and 21 cadaver heads by anterior rhinoscopy and videotaped rigid 30 degrees endoscopy. The vomeronasal organ was identified in 112 patients (32%) and in 8 cadaver heads (38%). The location, shape, type, and relation to sex of the vomeronasal organ were described. Ten specimens were examined histologically and histochemically for neuron-specific enolase (anti neuron-specific enolase), high-molecular-weight cytokeratin (anti-high-molecular weight cytokeratin), and low-molecular-weight cytokeratin (anti-low-molecular weight cytokeratin). Considering its variability in shape and the lack of immunohistochemical characteristics of nerve tissue, the present results are not suited to argue for functional significance of the vomeronasal organ in human beings. PMID- 15346009 TI - BMP-9-transduced prefabricated muscular flaps for the treatment of bony defects. AB - Autologous bone grafting techniques involve the use of tissues that need to be extracted from healthy sites. This can lead to significant donor site morbidity that causes a one-site defect to become a two-site defect. Bone grafts can be especially difficult to manipulate, because bone is a relatively nonmoldable tissue. Furthermore, the inability of a bone graft to contain a transplantable vascular supply also limits the possible size that such a bone graft can be. Because of these limitations, a graft that was moldable with a vascular supply would possess significant advantages in reconstructive applications. In this research, gene therapy techniques were used to create such a graft. An adenovirus expressing BMP-9 was injected into the latissimus dorsi of a nude animal to cause bony differentiation of that muscle. Differentiation of the muscle to cartilage in bone was measured by reverse transcription polymerase chain reaction and immunohistochemistry to determine the optimal time of flap elevation. After injection of the BMP-9 virus, the animals were biopsied weekly over a 3-week period. Both bone and cartilage markers were discovered in these tissues over the study period. Optimal flap elevation time was established to be 2 weeks after injection of the virus. PMID- 15346011 TI - Volumetric assessment of the distracted human mandible. AB - The mandibles of five patients who underwent unilateral or bilateral distraction osteogenesis were analyzed using computed tomography scans. The mandibles were reconstructed in three dimensions using General Electric computed tomography software. The total volume of each hemimandible was determined before and after distraction and compared. The mandibles were then segmented at 1-cm intervals, and the volumes of the segments were determined. The volumes of the mandibular segments before and after distraction were compared to determine the distribution of new bone through the mandible and the quantity of bone generated by distraction. The distracted hemimandibles increased in total volume by an average of 19.9% (statistically significant by the Student paired t test; P < 0.0001), whereas the nondistracted hemimandibles increased in total volume by an average of 5.2%. The segmental volumes of the distracted mandible were of similar or greater volume when compared to the preoperative mandibular segments. In addition, the distribution of regenerate bone mirrored the physiological distribution of the preoperative mandibular bone. This study indicates that distraction not only creates new bone but distributes that bone through the remodeled mandible in an anatomical pattern similar to that of the preoperative bone. PMID- 15346013 TI - In vitro analysis of the elution of tobramycin from a calcium sulfate bone void filler. AB - The use of synthetic calcium-based bone replacements has a recent history in craniofacial surgery as a non- to minimally load-bearing graft substitute. Given its avascular nature at the time of placement, infection is an ever present risk. Antibiotics are often added to the material during surgery based on an empirical impression. Whether effective antibiotic release actually occurs and over what time are not known for many of the available calcium-based preparations. The purpose of this study was to investigate the release kinetics of tobramycin from molded cylinders of Calcigen S bone void filler (Biomet, Warsaw, IN) and to compare this with the release kinetics of tobramycin from plaster of Paris-based systems as reported in the literature. Calcigen S bone void filler is a form of calcium sulfate that is closely related to plaster of Paris yet remains distinct. In vitro studies as well as clinical series have demonstrated that plaster of Paris is an effective vehicle to deliver tobramycin in which therapeutic doses of the antibiotic are released after implantation. In vitro analysis of the elution of tobramycin from Calcigen S bone void filler substrate is similar to that of plaster of Paris. PMID- 15346014 TI - Alopecia treatment with scalp expansion: some surgical fine points and a simple modification to improve the results. AB - In the current study, authors present their clinical experience with the esthetic reconstruction of alopecia by means of a tissue expansion technique in 74 consecutive patients who were treated between May 1986 and June 2002 in the Department of Plastic and Reconstructive Surgery. The principles of the conventional technique are mentioned briefly, but the authors essentially tried to explain a number of surgical fine points together with some simple modifications so as to get the maximum profit from the expanded tissue and to decrease the complication rate. In the first 39 patients of this study, who were treated with a conventional tissue expansion technique, the major complication rate was found to be 15.4%. In the last 35 patients, this rate was found to be decreased to 5.7%. The improvement in the major complication rate is attributed to the authors' surgical modifications compared with the conventional technique. The article provides a supplement to the existing literature, underscoring the importance of some surgical fine points and outlining a systematic way of planning expander placement and tissue expansion. PMID- 15346015 TI - Free flap transfer for closure and interposition-arthroplasty in noma defects of the lateral face associated with bony ankylosis. AB - Noma defects of the anterolateral face are often associated with fibrous or bony ankylosis fusing the mandibula to the skull base. According to the extent of the ankylosis, the temporomandibular joint mobility can be restricted or even completely frozen. In third world conditions the surgical approach to severe forms of bony ankylosis consists of a single linear opening osteotomy (trismus release) and the closure of the noma defect with locoregional flaps. Relapse of jaw immobility is common and may be caused by minor bone resection, the lack of adequate postoperative physiotherapy, or even the scarring of the defect coverage. In 4 years the authors have gained increasing experience with folded free flaps for simultaneous closure of outer and inner lining of large noma defects and the maintenance and training of re-established jaw function by the use of a dynamic external distractor fixed between the zygoma and the mandibular body. The authors report the bony reankylosis can be reduced by extended wedge osteotomies of the bony bridge and tip-like shaping of the ascending mandibular ramus. To preclude the reossification of the osteotomy site and fibrous scar formation, a dermofatty or muscular tail of the free flap is interposed into the bone gap. Two cases were treated according to this concept with a free parascapular and a latissimus dorsi flap in combination with simultaneous arthroplasty. During a 6-month follow-up period, no signs of a recurrent reduction of mandibular movement were noted in either case. PMID- 15346017 TI - Surgical treatment of ossified cephalhematoma. AB - Cephalhematoma is a collection of blood beneath the periosteum of the bones in the cranial vault. The treatment strategy of most cephalhematoma is conservative. However, in cases of ossified cephalhematoma causing deformities of skull, surgical management is indicated. From 1996 to 2002, the authors treated three cases of ossified cephalhematoma with the suggested pathogenesis of ossification, which cephalhematoma goes to ossification process, rather than calcification. Surgical treatment, which is bony shaving with a burr on the irregular margin site, was performed in one case in which the thickness of the skull in the bulging area was the same as in the normal area in preoperative computed tomography (CT) scan, and a depression did not exist in the operative field. In two cases in which there was any difference in bony density in the preoperative CT scan and depression after removal of cephalhematoma, the bony cap was remodeled into multiple pieces and the depressed region was reconstructed with a remodeled bone cap as an onlay bone graft. There was no evidence of complications and recurrence, and there was good reconstitution of the skull contour in all cases. PMID- 15346018 TI - Selective neurectomy of the masseteric nerve in masseter hypertrophy. AB - Asian women prefer to have more delicate and feminine features rather than prominent squared mandibular angles. It was proposed that a selective neurectomy of the masseteric nerve might result in muscular atrophy and subsequent reduction of the hypertrophic mass of the masseter muscle. This study is aimed at an elucidation of the precise course of the masseteric nerve and the impact of a neurectomy in case of masseter hypertrophy. Eleven heads of Korean cadavers were halved and dissected. The masseteric nerve and artery in the mandibular notch were clearly located. A selective neurectomy of the masseteric nerve was performed in four cases of masseter hypertrophy. The masseteric nerve and artery behind the temporalis muscle at the mandibular notch cross over to the masseter. The masseteric nerve was found 11.3 +/- 2.6 mm horizontally from the anterior border of the ramus and 11.3 +/- 2.6 mm above the mandibular notch and the superior masseteric artery, 25.6 +/- 3.7 mm from the ramus, and 7.8 +/- 2.6 mm above the notch. A neurectomy of the masseteric nerve can be used to reduce the hypertrophied masseter and as an ancillary procedure of mandibular angle ostectomy. One procedure can accompany another in the reduction of a prominent mandibular angle. PMID- 15346019 TI - Transmaxillary osteogenesis distraction combined with orthopedics and orthodontics in the correction of a severe retrusion of the upper maxilla. AB - A patient having adequate records and diagnosed as having Binder syndrome is presented. Nasomaxillary hypoplasia requires a definitive treatment, use of bone grafts, upper maxillary osteotomies, and advancement or a combination of both. Bone grafts can be reabsorbed, and complete maxillary advance modifies normal occlusion in a certain way, because the posterior sector is not compromised. The aim of this case treatment is to create a new osseous surface that makes the eruption of the permanent dental pieces easier and gives the correct skeletal position to the upper maxilla with distraction osteogenesis combined with an orthopedic appliance, transmaxillary segment osteotomy, and subsequent orthodontics. The absence of the osseous surface in the upper maxilla and the presence of supernumeraries in the anterior region determined the permanence of most of the temporary dental pieces, resulting in the impossibility of making the exchange to permanent dental pieces. The technique and the devices are simple and easy to manipulate. After the treatment, an adequate dental relation is restored as well as effective advance of the upper maxillary bone; thus, an excellent functional and stable esthetic result is achieved, avoiding any complication of velopharingeal incompetence. PMID- 15346020 TI - New regenerative surgical treatment of cystic diseases of the jaw by utilizing grafting of cancellous iliac bone and replanting of patient's teeth. AB - The authors developed a new regenerative surgical technique for cystic diseases of the jaw. In this surgery, all teeth that are rooted in or located adjacent to the cyst are extracted for replanting, and attached soft tissues, including cystic wall as well as dental pulp, are completely removed. Gingiva is cut open at the alveolar crest and abraded subperiosteally, the jaw on the cystic lesion is widely exposed, and cortical bone on the frontal wall of the cystic lesion is dissected. After the entire cyst is exposed, it is completely removed with scrapers and bars. The defect is reconstructed by the following procedures: the extracted and treated teeth are replanted; cancellous iliac bone (bone marrow) is grafted around the teeth and in the defect; and after thorough washing and making numerous small holes, the frontal wall of the cortical bone is returned to the original position and fixed. Three to 4 months later when the jaw has regenerated and the replanted teeth have taken, a prosthesis is set on the replanted teeth and biting is started. This technique was applied to a 16-year-old male patient who had odontogenic keratocysts on the lower right teeth (numbers 6, 5, 4, 3, 2, and 1), the lower left teeth (numbers 1, 2, 3, and 4), and the upper right teeth (numbers 5, 4, 3, and 2). The patient regained normal biting capability, and the esthetic outcome was also satisfactory. This technique would be a useful treatment method for cystic diseases of the jaw. PMID- 15346021 TI - Facelift approach with a hybrid SMAS rotation advancement flap in parotidectomy for prevention of scars and contour deficiency affecting the neck and sweat secretion of the cheek. AB - Tumors of the parotid gland are generally be removed by the standard external bayonet-shaped incision approach without reconstruction of the parotid bed. The disadvantage of this approach is frequently an obvious scar affecting the neck and a conspicuous hollow contour around the angle of the mandible in addition to a sweat secretion of the cheek (Frey syndrome). To overcome these disadvantages, especially the facial depressed deformity subsequent to parotid surgery, during the last several years, the author has concentrated on facelift incision used in combination with a hybrid SMAS rotation advancement flap. Twelve patients (7 male; 5 female) ranging in age from 32 to 73 years (mean age, 57.8 years) fulfilled the selection criterion of having a clinically benign discrete parotid lump with a benign preoperative fine-needle cytology result. Parotidectomy was performed using the modified facelift incision in conjunction with the rotation advancement hybrid SMAS flap. All patients were followed up every 3 months during the first year. During follow-up, the patients were specifically asked about their satisfaction with their postoperative appearance and whether they would consent to the operation again. The vascularized hybrid vicryl mesh/SMAS rotation advancement flap is clinically simple to perform and provides satisfactory cosmetic and functional results in patients undergoing conservative parotidectomy and prevents the gustatory sweating. There are no drawbacks to the use of the modified facelift incision to remove tumors of the parotid gland. PMID- 15346022 TI - The effect of timing of surgery for velopharyngeal dysfunction on speech. AB - The timing of surgery for velopharyngeal dysfunction has been based on assumptions about the relation between age, speech development, and velopharyngeal dysfunction. Cleft palate teams often counsel parents to have an intervention for velopharyngeal dysfunction performed earlier rather than later, believing that earlier interventions result in more rapid or better normalization of speech. The objective of this retrospective chart review study is to determine whether the age at surgical intervention for velopharyngeal dysfunction has an effect on the subsequent length of speech therapy. Of 174 patients included in the study database, 36 had velopharyngeal dysfunction for which further velopharyngeal management was required. Of the 36 patients who received surgical velopharyngeal dysfunction management, 27 had verifiable speech therapy records. These 27 patients represent the study population. The outcome measure was the total length of subsequent speech therapy until speech normalization. The data suggest that there is no relation between the age at velopharyngeal dysfunction surgical management and the amount of speech therapy needed to achieve normalization of the speech impairments secondary to velopharyngeal dysfunction after that management. In conclusion, 1) the age at surgical velopharyngeal dysfunction management (pharyngeal flap or sphincter pharyngoplasty) does not have an effect on subsequent normalization of speech as measured by the duration of speech therapy necessary to achieve normalization of the speech impairments secondary to velopharyngeal dysfunction after that management, and 2) the age at surgical velopharyngeal dysfunction management does not affect the likelihood of subsequent surgical velopharyngeal dysfunction management procedures. PMID- 15346023 TI - Spring-mediated cranial reshaping for craniosynostosis. AB - The evolution of modern craniofacial surgery has come full circle from the early strip craniectomies to the complete calvarial remodeling and now back to attempts at minimally invasive surgical interventions. The goal of the craniofacial surgeon has always been the correction of form and function with minimization of associated morbidity and mortality. Particularly problematic has been the ability to maintain the anatomical correction beyond the result seen on the operating room table secondary to changes with growth. The ability to improve the clinical result in a growing and developing child has been the impetus for dynamic treatment modalities. Dr Claes Lauritzen's pioneering work in this area has been particularly successful using internal springs to correct craniofacial deformities. The purpose of this study is to assess this treatment modality clinically in terms of safety and efficacy and to develop a methodology for the spring formation that would be easily reproducible. This is an institutional review board-approved prospective study of 15 children (11 male, 4 female) with non-syndromic sagittal suture synostosis. All patients were treated with a sagittal strip craniectomy and placement of 2 omega-shaped stainless steel springs at a mean age of 3.9 months. Patients were followed clinically and with cephalograms; after reossification of the intervening bone, the springs were removed at a mean age of 8.2 months. The mean force applied at initial placement of the springs was 6.9 N, and the mean spring deflection at formation was 6.87 cm. All patients completed the study protocol without any significant morbidity or any mortality. Perioperative variables, including blood loss, transfusion rate, operative time, intensive care unit stay, hospital stay, and hospital charges, were all significantly less (P < 0.05) in this study group compared with children with the same diagnosis treated with cranial vault reshaping during the same period. Furthermore, the preoperative mean cephalic index of 64.3 corrected to 77.6 after surgery and was maintained over time. Spring-mediated cranial reshaping is efficacious and safe for the treatment of sagittal synostosis. Long term study of cranial development and clinical morphology are ongoing to validate further the effectiveness of this treatment modality. PMID- 15346025 TI - Outcomes of tongue-lip adhesion for neonatal respiratory distress caused by Pierre Robin sequence. AB - Tongue-lip adhesion can be effective in relieving tongue-based airway obstruction in the neonatal period. However, longitudinal evaluation of these patients demonstrates they require additional procedures for ultimate control of their airway, feeding, and orthognathic relationship. Charts of patients with Pierre Robin sequence who underwent unsuccessful nonoperative treatment and tongue-lip adhesion procedures by the senior author between 1989 to 1999 were reviewed. In particular, secondary interventions were quantified and qualified to determine if tongue-lip adhesion was a definitive treatment of the pathology caused by this sequence. Eleven patients with ages ranging from 2 to 6 weeks underwent initial tongue-lip adhesion. Seven patients were syndromic (3 Stickler's, 1 Goldenhar's, 1 Fragile X, 1 Miller's, 1 chromosome 15 rearrangement), and four had isolated Pierre Robin sequence. Two patients had dehiscence of their initial repair within 2 weeks, for a primary surgical success rate of 82%, consistent with other published reports. A detailed follow-up of these patients revealed that 10 of 11 (91%) required additional intervention for airway or feeding issues. Of those requiring secondary intervention, an additional 1.9 secondary procedures were performed per patient to achieve control of the airway, feeding, and orthognathic problems. More than half of the patients needed gastrostomy tubes to overcome feeding difficulties. Five patients necessitated secondary surgery for recurrent airway obstruction within 4 months of the initial surgical treatment (four distractions and one repeat tongue-lip adhesion). Four others required distraction at an older age for orthognathic indications. Two patients were successfully treated with tongue-lip adhesion only. Tongue-lip adhesion has a high initial success rate for correction of neonatal airway obstruction. However, long-term follow-up indicates a high incidence of secondary intervention requirements. In retrospect, perhaps tongue-lip adhesion should be considered a temporizing procedure for most patients with Pierre Robin sequence. PMID- 15346026 TI - Genetic profiling of granular cell myoblastoma. AB - Granular cell tumor (GCT), or granular cell myoblastoma, is a relatively uncommon lesion of the soft tissues. It can occur in any organ, and the tongue is more often affected. GCT has unknown etiology, uncertain histogenesis, and a not always benign nature. Benign myoblastomas are the great majority, but rare malignant lesions have been reported. To have more information regarding the genetic events involved in GCT, the authors decided to perform an expression profile. A sample was derived from a surgically resected GCT of the tongue. RNA extracted from normal tongue (mucosa plus muscle) was used as control. By using DNA microarrays containing 19,200 genes, the authors identified several genes for which expression was significantly up- or down-regulated. The differentially expressed genes cover a broad range of functional activities: (1) signal transduction, (2) cell cycle regulation, and (3) cytoskeleton organization. It was also possible to detect some genes whose function is unknown. The data reported are, to the authors' knowledge, the first genetic portrait of GCT. Mutations in some of the described genes are related to neural alterations and mental diseases, and this fact supports the idea of a neural origin of myoblastoma. Several markers have been identified that will help in identifying the biological behavior (when malignant lesions will be described), as well as the gene whose products could be potentially disease-specific targets for therapy. PMID- 15346027 TI - In search of causative factors of deformational plagiocephaly. AB - This is a preliminary study of 37 infants with deformational plagiocephaly. In this first effort to create a profile of factors that may contribute to the onset and progress of developmental plagiocephaly, parents were interviewed concerning selected practices before and immediately after the birth of their infants. Results of the study do not reveal a clear or concise pattern of factors that may contribute to this condition. There needs to be additional research on the benefits and potential hazards of sleep position to infants as one of the factors that may contribute to deformational plagiocephaly. PMID- 15346028 TI - Do we need three-dimensional computed tomography in maxillofacial surgery? AB - In a retrospective clinical study (2000-2003), 121 patients' radiographs and computed tomography scans were reviewed to establish the clinical value of three dimensional computed tomography. Eighty patients had computed tomography scans; 48 had three-dimensional computed tomography scans for diagnosing facial fractures, 3 for diagnosing temporomandibular joint ankylosis, 1 for tumor with bone destruction, and 1 for a mandibular cyst. It is concluded that axial, coronal, and three-dimensional computed tomography is of crucial importance and should be mandatory for all suspected comminuted and displaced midface fractures instead of plain radiographs. Three-dimensional computed tomography is also recommended for comminuted mandibular fractures and temporomandibular joint ankylosis. Three-dimensional computed tomography is not recommended for the diagnosis of minimally displaced fractures. PMID- 15346030 TI - Cervical impalement injury. AB - Impacted injuries of the head and neck are uncommon. Associated injuries can be present and can be a serious problem. We present an unusual clinical case and discuss the management of this complex injury. Head and neck injuries deserve special attention because of their enormous functional and esthetic significance. Contusion, abrasion, retained foreign bodies, laceration, and avulsion are some of the most frequent injuries. Impalement injuries are not so common. These lesions combine aspects of blunt and penetrating trauma and usually result from penetration by a large, rigid, blunt-tipped object that traverses a certain body area in a through-and-through fashion and often remains in situ at the time of presentation. There have been many case reports describing these injuries in the trunk and extremities; however, cases of head and neck impalement are still lacking in number. A case of an unusual cervical impalement is presented, together with a discussion of the management of this complex injury. PMID- 15346031 TI - Ossifying fibroma of the skull: interactive image-guided minimally invasive localization and resection. AB - Ossifying fibroma is a benign fibro-osseous tumor commonly affecting the craniofacial bones. It is considered to be a locally aggressive and quickly expansible bone lesion. Because of its aggressive nature and high recurrence rate, early detection and complete surgical removal are essential. Usually, these lesions are excised extensively by craniectomy, and bone loss is reconstructed by cranioplasty using acrylic resin or titanium implants. Alternatively, in the management of skull-ossifying fibroma, an image-guided technique using surgical navigation may provide precise information about localization, enabling complete removal, thereby operating with minimal exposure and within narrow resection borders and avoiding significant bone deformity. A 39-year-old male patient with a history of renal cell carcinoma was admitted to our hospital because a radionuclide scintigraphic bone scan revealed increased uptake in a small area located at the left lateral skull bone. The high-resolution computed tomography scan showed that the lesion was located inside the diploe, destroying the inner table of the calvarium. The patient underwent minimally invasive bone lesion removal using an interactive image-guided approach. Complete resection of the neoplastic lesion was achieved. The histopathological examination revealed an ossifying fibroma. The postoperative course was uneventful, and the patient was discharged 3 days after intervention. To date, there has been no evidence of local recurrence. Interactive multimodal planning and intraoperative image guidance offer an interesting approach for biopsy and minimally invasive removal of small ossifying fibroma lesions of the skull, especially in less accessible locations. PMID- 15346032 TI - Orthognathic surgery: an informed consent model. AB - In this study, the authors propose an informed consent form for orthognathic surgery. A careful review of the international literature and clinical practice suggested the feasibility of dividing the informed consent form into two parts. In first part, the diagnostic procedures and the therapeutic and surgical times are described. The patient must sign it as soon he or she accepts the orthodontic/surgical therapy proposed. The second part describes the possible problems and difficulties of the procedure, and it must be read by the patient before starting the therapy. In the opinion of the authors, this informed consent form allows the patient to know the risks related to the orthodontic/surgical therapy, thus preserving the surgeons from the civil risk and penalties of omission. PMID- 15346033 TI - Unusual thyroid gland vascularization. AB - During a dissection course, an unusual vascularization of the thyroid gland was found in a 69-year-old human male cadaver. On the right side, two arteries from the thyrocervical trunk supplied the lower half of the right lobe of the thyroid gland: the inferior thyroid artery and an additional vessel with a main trunk, found to be split into an upper and lower branch at the level of the longus colli muscle. The upper branch supplied the inferior constrictor pharyngeal muscle and the esophagus, whereas the lower branch coursed, at first, onto the trachea and finally leaving it to anastomose with the inferior thyroid artery. PMID- 15346034 TI - Situs inversus totalis with accompanying craniodiaphysial dysplasia: a new syndrome? AB - A 20-year-old man with craniofacial anomalies of craniodiaphysial dysplasia (CDD), facial paralysis, mental retardation, and situs inversus totalis is described. Similar features except situs inversus totalis are also present in two of his sisters. The authors believe this to be the first reported case of CDD with accompanying situs inversus totalis. PMID- 15346035 TI - Outer table craniotomy for frontal sinus mucocele. AB - Variants of the external osteoplastic flap procedure or endoscopy have been used to approach the frontal sinus mucocele. The authors introduce a modified external approach for radical resection of the mucocele. Using the bicoronal skin incision, the skin flap exposed the right upper orbital rim. The outer table craniotomy was then performed to expose the frontal sinus cavity while carefully preserving the inner table, with radical removal of the mucocele mucosa. The sinus cavity was irrigated with hyper-oxide solution to ensure adequate destruction of possible residual mucosa. The fascia of the frontalis muscle was split, with one part placed into the mucocele cavity to plug the nasofrontal duct. The advantages of the procedures for mucocele removal include clear visualization of the frontal sinus for radical resection of the mucosa, preservation of the inner table avoiding dura manipulation, prevention of central nervous system infection, possibly lower rates of recurrence, prevention of mucosal ingrowth by plugging of the nasofrontal ducts with fascia, and favorable cosmetic outcome. The disadvantages are more intensive surgery comparable to the endoscopic approaches and execution difficulties when the frontal mucocele is small. Additional clinical studies are needed to evaluate the efficiency and safety of this procedure. PMID- 15346037 TI - A reflectable case of obstructive sleep apnea in an infant with Crouzon syndrome. AB - Obstructive sleep apnea has recently drawn attention as a cause of sudden death among infants. Life-threatening obstruction of the upper airway is encountered in patients with syndromic craniosynostosis. Early definitive management of obstructive sleep apnea can conquer this critical situation. Although early tracheostomy can solve the problem, successful early midfacial distraction has been reported. In this report, a reflectable case of sudden death caused by a severe obstructive sleep apnea attack at home just before the midfacial distraction, during the waiting period for the surgery of midfacial distraction, is described. The authors stress the importance of preoperative care of the upper airway and the early definitive treatment using distraction osteogenesis for midfacial hypoplasia in infantile syndromic craniosynostosis. PMID- 15346039 TI - Treatment of temporomandibular joint ankylosis in children: is it necessary to perform mandibular distraction simultaneously? AB - Temporomandibular joint (TMJ) ankylosis in children disturbs not only mandibular growth, but also facial skeletal development. Costochondral graft was used to ensure growth, but it had proven to be unpredictable. The authors evaluate retrospectively 41 patients who underwent temporomandibular joint reconstruction during the last 10 years. Twenty were treated by costochondral graft, 15 by arthroplasty, and 6 by other surgical procedures, and they were excluded. The etiology was septic in 54% of the cases. Follow-up was at least 12 months in all cases. Arthroplasty was a quicker and easier procedure than the costochondral graft, reducing operating time, risk of blood transfusion, and hospital stays and costs. It also was associated with less risk of reankylosis, 13%vs 25%. Furthermore, it was associated with a minor morbidity and secondary complications. Seventy-five percent of the patients treated with bone graft required additional secondary surgery. Radiographically, the authors observed a remodeled neocondyle at the level of proximal mandibular end in cases treated by arthroplasty. On clinical examination, patients showed variable degrees of facial deformity and an unknown potential of mandibular growth after TMJ arthroplasty. The authors also observed improved clinical and radiologic appearance after ankylosis correction. Is it reasonable to perform ankylosis release and mandibular distraction simultaneously without knowing which patients will be able to experience growth with time? In that case it would be necessary a predict growth to apply the exact amount of mandibular distraction for obtaining stable results. Timing of mandibular distraction, after TMJ arthroplasty is performed and mandibular function restored, must be specific to each patient's needs, assuring the best distraction conditions and planning. The authors present their treatment protocol, including TMJ joint arthroplasty with temporal muscle interposition, and mandibular distraction osteogenesis, as a second procedure, to correct residual asymmetry or retrognathism if necessary. PMID- 15346041 TI - Fronto-orbital mucoceles: our experience. AB - Frontal sinus mucoceles are rare benign neoplasms that can result in bony erosion extending from the borders of the sinus into the orbital cavity. The authors report the fronto-orbital mucoceles they have observed in the last 8 years. The authors used an "open surgery" approach in 12 fronto-orbital mucoceles, characterized by an osteoplastic frontal flap, through a coronal incision or Lynch incision. In this way, they were able to expose the frontal sinus and to remove completely the mucocele from the periorbita. Only one recurrence is reported 2 years after surgery. Excellent aesthetic results have been reached in all cases. In the literature, many different positions are reported for the treatment of mucoceles. The endoscopic surgery of the paranasal sinuses has become the procedure of choice for mucoceles of maxillary, ethmoidal, and sphenoidal sinuses. However, few authors have recommended the endoscopic approach for frontal mucoceles. The indications and limitations of endoscopic and "open" surgery are critically discussed in the light of the authors' personal experience and current literature. PMID- 15346042 TI - Improved esthetic results with fine-tip Dermabond application technique. AB - Tissue glues and adhesives have achieved increasing popularity as alternatives to small wound closure. When applying these substances, it is often difficult to avoid contact with the surrounding skin and foreign objects such as surgical gloves. A technique for the application of Dermabond is described in this report. Twenty wounds less than 10 cm in length were reapproximated using a fine-tip tuberculin syringe applicator. The wounds were evaluated immediately after the application, and 2, 4, and 6 weeks after surgery. Use of the tuberculin fine-tip technique allowed reapproximation of the wound edges with no clinical evidence of surrounding tissue damage. There were no apparent decreases in wound strength or associated discoloration or fuzziness onto the skin. Esthetic results were considered good to excellent by patients. Dermabond can be accurately applied with a tuberculin syringe, avoiding the surrounding tissue damage and foreign object adhesion reported in the literature. PMID- 15346043 TI - Changing ethical standards in scientific publication. PMID- 15346045 TI - The principles and technique of resection of soft parts for sarcoma. 1958. PMID- 15346046 TI - Pioneers in musculoskeletal oncology: Musculoskeletal Tumor Society Founders Lecture. AB - I was invited to deliver the third annual Musculoskeletal Tumor Society's Founder's Lecture, at our annual meeting on May 2, 2003 in Chicago, IL. The Musculoskeletal Tumor Society began in 1977 when a small group of physicians, mainly orthopaedic surgeons, met to discuss ways of improving the outcome of the rare and usually fatal primary malignant neoplasms of bone. Treatment of patients with these types of cancers most often was amputation. Initial criteria for membership included participation in group clinical studies. As individuals and collectively as a tumor society, we have witnessed and participated in a revolution in the care of these patients resulting in markedly increasing survival from less than 20% to nearly 70% and dramatically lessening disabilities by using limb-sparing surgery instead of amputation. I have elected to present brief descriptions of some of the individuals and their contributions that have led to these noteworthy improvements in patient care and final results. These individuals lived in different countries during different times and are from various fields of medicine. The time constraints imposed by the lecture have made it impossible to include all the deserving pioneers. PMID- 15346047 TI - Treatment of advanced metastatic lesions of the acetabulum using the saddle prosthesis. AB - Current methods of treating advanced patients with metastatic periacetabular disease are complex and result in high complication rates. The purpose of this study was to show whether the implantation of the saddle prosthesis would serve as an additional tool to help treat metastatic disease in these patients. From 1991 to 2003, 20 patients with advanced metastatic periacetabular lesions (Harrington Class III) were treated using the saddle prosthesis. Goals of surgery were a decrease in pain, functional restoration, and ambulation. The mean age was 61 years. Average length of followup was 20 months. Postoperatively, ambulation was achieved in 16 of 20 patients. There were four postoperative complications (20%) in three patients. Surgical goals were met in 18 of 20 patients. The MSTS ISOLS emotional score was 2.9 of 5. The average total MSTS-ISOLS score was 16.6 of 30 (55%). Using the Allan scoring system consisting of analgesia, independence and ambulation, and mobility, all scores had significant improvements postoperatively. Careful surgical indications and technique should result in a stable, functional reconstruction allowing patients the ability to ambulate outside the house with a cane. Patients can expect to be emotionally satisfied with the procedure while using nonnarcotic analgesia and can expect an improved quality of life despite bone metastasis. PMID- 15346048 TI - VEGF expression in osteosarcoma correlates with vascular permeability by dynamic MRI. AB - Dynamic enhanced magnetic resonance imaging has been used to assess tumor angiogenesis in osteosarcoma. Vascular endothelial growth factor has been shown to correlate with pulmonary metastasis and a poor prognosis in osteosarcoma. The purpose of this investigation was to determine whether vascular endothelial growth factor expression in osteosarcoma correlates with vascular permeability detected by dynamic enhanced magnetic resonance imaging and to explore the role of dynamic enhanced magnetic resonance imaging as a noninvasive means of assessing tumor angiogenic activity. Fifty-five osteosarcoma patients with osteosarcoma enrolled in a treatment protocol that included dynamic enhanced magnetic resonance imaging. In 15 patients, tumor tissues were available for vascular endothelial growth factor immunohistochemical studies. A two-compartment model used the exchange rate constants (kep) between the plasma and tumor compartments to quantify vascular permeability during dynamic magnetic resonance imaging studies. Immunohistochemical staining for vascular endothelial growth factor was graded according to the intensity and number of positively stained cells. Vascular endothelial growth factor-positive tumors showed higher mean vascular permeability when compared with vascular endothelial growth factor negative tumors. Vascular permeability also correlated with increasing vascular endothelial growth factor expression. The preliminary results in this study show an association between vascular endothelial growth factor and dynamic MR signal enhancement in osteosarcoma. Dynamic enhanced magnetic resonance imaging should be investigated as a means to prognosticate osteosarcoma patients with osteosarcoma according to their tumor angiogenic activity. PMID- 15346049 TI - Dog osteogenic sarcoma microvasculature observed with a Spalteholz technique. AB - A well-distributed, patent microvascular network is essential for adequate, uniform delivery of chemotherapy into solid tumors. This network has not been evaluated in osteogenic sarcoma. Spalteholz tissue clarification was used to observe the microvasculature of canine humeri bearing osteogenic sarcoma. Freshly amputated limbs, obtained from therapeutic amputation, were infused with a micron sized carbon particle solution, frozen, and then cut into sagittal and axial 0.5 mm thick sections. They were photographed, then radiographed using high resolution Faxitron xray, chemically treated to clarify the tissue, and then rephotographed. Microvasculature was identified by the localization of carbon particles, which were unaffected by the clarification process, within the clarified sections. Clarified section photographs were digitized to gray scale levels and analyzed using IMAGE software; levels are directly related to capillary density. Faxitron and original images were registered to the clarified images to identify tissue regions. Multiple regions of interest from normal muscle, fat, bone, and tumor regions were selected and averaged. The microvasculature of the tumor was inhomogeneous, whereas its density was considerably lower than normal adjacent muscle and bone (range, 56-72% lower). These findings suggest that insufficient microvascular density and distribution may provide additional explanation for the poor response of solid tumors to chemotherapy and radiation therapy. PMID- 15346050 TI - Functional outcome of endoprosthetic proximal femoral replacement. AB - Endoprosthetic proximal femur replacement is a well-accepted method for treatment of primary bone tumors; however the functional results of treatment are not well documented. To evaluate functional outcomes, we recorded the Toronto Extremity Salvage Score and Musculoskeletal Tumor Society 1987 and 1993 scores in 29 patients, and also recorded Musculoskeletal Tumor Society scores alone in four more patients treated with endoprosthetic proximal femur replacement. The mean followup was 3 years. Twelve patients had a total hip endoprosthetic proximal femur replacement, and 21 had a bipolar hip endoprosthetic proximal femur replacement. In nine patients, the greater trochanter was attached to the femoral prosthesis. Sixteen patients had an abductor soft tissue repair, and in eight patients, no abductor repair was possible. The mean Musculoskeletal Tumor Society 1987 score was 23.2 +/- 4.1 points of 35 points. The mean Musculoskeletal Tumor Society 1993 score was 67.7 +/- 12.0%. The Toronto Extremity Salvage Score mean was 76.2 +/- 16.2 points of 100 points. Functional scores did not differ significantly between abductor repair types. There was a trend toward less disability in patients with abductor soft tissue repair compared with patients with no abductor repair. Functional results were similar in patients receiving bipolar and total hip replacements. PMID- 15346051 TI - Kyphoplasty enhances function and structural alignment in multiple myeloma. AB - We prospectively evaluated 19 patients with multiple myeloma who had kyphoplasty for vertebral compression fractures. Functional status was assessed preoperatively and 3 months postoperatively using the Oswestry Disability Index. Restoration of anterior and midvertebral height was assessed using lateral radiographs. Meaningful improvement occurred in 16 of 19 patients, with a reduction of the average Oswestry Disability Index from 49 +/- 16.6 to 32.6 +/- 13.6. Partial restoration of anterior vertebral body height was achieved in 76% of levels with an average of 37.8% restoration of the defect. Partial restoration of midvertebral body height was achieved in 91% of levels with an average restoration of 53.4% of the defect. There were no significant complications. These results were compared with results of a cohort of 26 patients with osteoporotic compression fractures treated with kyphoplasty at 37 levels. There was no difference between the groups in terms of Oswestry Disability Index improvement and midvertebral height restoration after 3 months. Greater anterior vertebral height restoration was achieved in the osteoporotic group (51.2% versus 37.8%). Kyphoplasty is a safe treatment modality for myeloma-related vertebral compression fractures. Efficacy in terms of pain relief and functional outcome is comparable with the results in patients with osteoporosis. PMID- 15346052 TI - Gene expression in cancer cells is influenced by contact with bone cells in a novel coculture system that models bone metastasis. AB - Contact between bone cells and cancer cells (heterotypic cell contact) is thought to play a central role in the initial growth and progression of metastatic cells. Attempts at studying heterotypic contact in vitro and in vivo have been confounded by difficulty in controlling how and when heterotypic contact occurs between unlike cells. A novel model, the micropatterned coculture system, is described that quantifies and controls heterotypic contact between cancer cells and bone cells in vitro. The micropatterned coculture system is biocompatible, and is modified easily to accommodate two or more different populations of cells. Immunofluorescence of cocultures of prostate cancer-3 cells and osteoblasts show the precise control of cell interactions. Ribonucleic acid of sufficient quantity and quality is isolated readily from cells cocultured on the micropatterned coculture system. The expression of the metastasis associated genes urokinase plasminogen activator, insulinlike growth factor binding protein-1 and insulinlike growth factor binding protein-3 are regulated in response to heterotypic contact and soluble factors respectively. A model of bone metastasis based on the micropatterned coculture system technology will streamline the process for testing therapeutic agents, so that more molecules can be identified for animal and clinical testing at less cost and in less time than using conventional methods. PMID- 15346053 TI - hTERT Is highly expressed in Ewing's sarcoma and activated by EWS-ETS oncoproteins. AB - Although the fusion proteins EWS-ETS are uniquely associated with Ewing's sarcoma and have been shown to have transformational properties, they are not the only determinants of oncogenesis. Therefore, before molecular-based therapy can be initiated, a better understanding of the molecular network specific to Ewing's sarcoma is mandatory. Specimens from 31 patients with Ewing's sarcoma were analyzed immunohistochemically. We found that human telomerase reverse transcriptase was expressed highly (78%) in Ewing's sarcoma. The mean followup was 7 years (range, 1-21 years), and human telomerase reverse transcriptase expression was correlated with outcome. Because we did not find an association between expression pattern and survival, human telomerase reverse transcriptase may not serve as a tumor marker in Ewing's sarcoma. However, the human telomerase reverse transcriptase promoter is shown to be activated by the fusion proteins. Therefore, transcriptional regulation via EWS-ETS may account for the high human telomerase reverse transcriptase expression in Ewing's sarcoma. PMID- 15346054 TI - Survival data for patients with malignant schwannoma. AB - Malignant schwannomas are uncommon sarcomas that arise from the sheath of Schwann surrounding peripheral nerve fibers. These tumors may arise spontaneously in adult patients or may occur with reportedly increased frequency in patients with neurofibromatosis Type I. The tumors have had a reputation for malignancy with rapid metastasis, especially when they arise in relation to neurofibromatosis. We have reviewed the experience of the connective tissue oncology unit with patients with these tumors during the past 25 years. There were 80 malignant schwannomas identified in 41 females and 39 males with a mean age of 36 +/- 17 years. The largest numbers of tumors were seen in the thigh, shoulder, and pelvis but they also occurred in more central (abdomen and thorax) and distal (forearm, hand and foot) locations. The Musculoskeletal Tumor Society stages showed that the majority of the patients had Stage II (high grade but not metastasized) tumors; however, additionally, 10 of the patients had metastases at outset (Stage III). Patient gender was not a factor but stage was a notable determinant of survival. Anatomic location also had a significant effect. Overall, the entire series of patients had a survival of 85% at 11 +/- 5 years and even more remarkable was the fact that there was no difference between the numbers and outcome statistics for tumors arising spontaneously and those occurring in patients with neurofibromatosis. PMID- 15346055 TI - Long-term effects of connective tissue cancer treatment. AB - In 1999, we began a study to assess the long-term effect of connective tissue cancer treatment on clinical, social, and psychologic aspects of the lives of surviving patients. A specially designed computer program generated an 85-item questionnaire, which was sent to more than 2000 patients with malignant bone and soft tissue neoplasms. Twelve hundred forty-four patients responded. The data were entered into a computer system and were correlated with the clinical information already contained in the system for the individual patients. Although there are many possible uses for these data, we chose to do a study comparing the lifestyle and physical and sociologic problems for 144 patients treated with chemotherapy and surgery for high-grade osteosarcoma against a control population consisting of 61 patients treated surgically for benign giant cell tumors of bone. The data show a remarkable degree of compensation on the part of the patients with the malignant tumors in terms of some problems but some significant differences particularly in physical status and functional limitations. PMID- 15346056 TI - Inferior vena cava filters prevent pulmonary emboli in patients with metastatic pathologic fractures of the lower extremity. AB - The records of 47 consecutive patients with metastatic pathologic fractures of the lower extremity were analyzed with respect to thromboembolic complications. All patients were unable to receive pharmacologic deep venous thrombosis prophylaxis, and were stratified into two groups, based on use of an inferior vena cava filter. Group I (n = 24) consisted of patients who had an inferior vena cava filter plus mechanical deep venous thrombosis prophylaxis (compression stockings and sequential compression boots); Group II (n = 23) consisted of a group of patients receiving only mechanical deep venous thrombosis prophylaxis. All patients had routine lower extremity venous duplex imaging preoperatively, postoperatively, and before hospital discharge. At final followup, patients were examined for deep venous thrombosis and reviewed for thromboembolic events. At a mean followup of 11.5 months, Group I had two detectable deep venous thromboses and no pulmonary emboli; Group II had one detectable deep venous thrombosis and five pulmonary embolisms. In Group II, 40% (two of five) of pulmonary embolisms were fatal, yielding an 8.7% (two of 23) group mortality rate. Overall, the entire group had an approximately 17% deep venous thrombosis rate. Only 6.4% (three of 47) of deep venous thromboses were detectable by standard duplex imaging. The majority of deep venous thromboses (five of eight, 62.5%) were nondetectable by duplex imaging. Overall, a 4.3% (two of 47) death rate was attributable to pulmonary embolism. In contrast, an 8.6% (four of 47) mortality rate occurred in Group II alone. All pulmonary embolisms occurred in patients who did not receive an inferior vena cava filter. The majority of venous thromboses (62.5%) were not detectable on duplex scanning, therefore were thought to arise from the pelvic venous system. Complications related to inferior vena cava filter insertion were minimal. For patients with metastatic pathologic fractures of the lower extremities who are unable to receive pharmacologic deep venous thrombosis prophylaxis, the use of inferior vena cava filters, in conjunction with standard mechanical deep venous thrombosis prophylaxis, is a procedure that has a low risk and is useful adjunct to prevent fatal pulmonary embolisms. PMID- 15346057 TI - The incidence and prognosis of osteosarcoma skip metastases. AB - The poor prognosis for patients with osteosarcoma skip metastases has been established in the literature; however, most of the data in the literature concerning osteosarcoma skip metastases were collected before routine use of magnetic resonance imaging and before the use of modern multiagent chemotherapy regimens. This retrospective multiinstitutional study was done to evaluate the efficacy of magnetic resonance imaging to detect skip metastases and to evaluate the prognosis for these patients when treated with modern multiagent chemotherapy protocols. In a group of 155 patients who were younger than 26 years when diagnosed and treated for high-grade osteosarcoma of the long bones, 10 (6.5%) patients with skip metastases were identified. Eight of the 10 lesions were detected on pretreatment magnetic resonance imaging scans and two were detected only from the surgical pathology specimens. Five patients had concomitant pulmonary metastases at presentation and five patients had no other known site of diseases. All 10 patients died, with an average survival of 32.5 months (range 11 71 months). The five patients with only skip metastases at presentation survived an average of 27.2 months (range 15-44 months). Despite advances in the treatment of patients with osteosarcomas, those with skip metastases continue to have a poor prognosis, and they should be counseled regarding the limitations of current therapies. PMID- 15346058 TI - Intercalary femur and tibia segmental allografts provide an acceptable alternative in reconstructing tumor resections. AB - Intercalary femur and tibia segmental allografts were implanted in 59 consecutive patients after segmental resection-52 for malignant and seven for benign aggressive bone tumors. The patients were followed up for an average of 5 years. Allograft survival was determined with the Kaplan-Meier method. Infection, fracture, and nonunion rates were determined. The overall 5-year survivorship for the 59 intercalary allografts was 79%, and we found no significant differences between allograft survival in patients receiving or not receiving adjuvant chemotherapy. Infection and fracture rates were 5% and 7% respectively. From 118 host-donor junctions, 11 did not initially heal (9%). The nonunion rate (10 of 69 osteotomies) for diaphyseal junctions was higher than the rate (one of 49 osteotomies) for metaphyseal junctions. Although some patients required reoperations because of allograft complications, it seems that the use of intercalary allograft clearly has a place in the reconstruction of a segmental defect created by the resection of a tumor in the diaphyseal and /or metaphyseal portion of the femur or tibia. PMID- 15346059 TI - Bisphosphonates may reduce recurrence in giant cell tumor by inducing apoptosis. AB - Giant cell tumor of bone is an aggressive tumor characterized by extensive bone destruction and high recurrence rates. This tumor consists of stromal cells and hematopoietic cells that interact in an autocrine manner to produce tumoral osteoclastogenesis and bone resorption. This autocrine regulation may be disrupted by novel therapeutic agents. Nonspecific local adjuvant therapies such as phenol or liquid nitrogen have been used in the treatment of giant cell tumor, but specific adjuvant therapies have not been described. The bisphosphonates pamidronate and Zoledronate can induce apoptosis in giant cell tumor culture in a dose-dependent manner. We established giant cell tumor cultures from patients with extensive destruction of bone. One of the four cultures formed osteoclastlike giant cells in vitro after more than six passages without exogenous receptor activator of NF-kappaB ligand or macrophage colony stimulating factor. Annexin V staining, presence of active cleaved form of caspase-3, and disappearance of poly (ADP-ribose) polymerase on Western blotting indicated activation of apoptosis by bisphosphonates in giant cell tumor. These results indicate that topical or systemic use of pamidronate or zoledronate can be a novel adjuvant therapy for giant cell tumor by targeting osteoclastlike giant cells, mononuclear giant cell precursor cells, and the autocrine loop of tumor osteoclastogenesis. PMID- 15346060 TI - Combinations of radioprotectants spare radiation-induced damage to the physis. AB - Radiotherapy used in the treatment of bone and soft tissue sarcomas in pediatric patients often results in undesirable growth plate damage. Radioprotectants may hold promise in the selective protection of growth plate tissue in this setting. In an animal model, the hypothesis tested was that pentoxifylline, selenium, or misoprostol, used in combination with amifostine, would significantly reduce longitudinal growth loss during one radiation dose exposure to a greater extent than the protection provided by only amifostine without increased morbidity or mortality or adverse effects on bone mineral density. Amifostine alone and in combination with each of the other radioprotectants resulted in limb discrepancy reduction to levels significantly less than radiated controls. The tibial length discrepancy in the selenium and amifostine group was 12.1 +/- 0.8%, less than the 15.5 +/- 2.6% tibial length discrepancy in the animals treated with amifostine alone, and less than the mean 18.8% tibial length discrepancy in the radiated limbs without radioprotection. There were no adverse effects on bone density in any group, but the selenium and amifostine group showed some increased mortality. Combinations of amifostine with these radioprotectants show efficacy in growth plate radioprotection and therefore warrant additional study in a clinically relevant fractionated model. PMID- 15346061 TI - Telomerase reverse transcriptase subunit expression is associated with chondrosarcoma malignancy. AB - Expression of the telomerase reverse transcriptase subunit telomerase reverse transcriptase gene is associated with most human malignancies. Because telomerase reverse transcriptase is rarely expressed in normal tissue, its presence in pathologic specimens is considered a marker of transformed cells. Moreover, high levels of expression have been correlated with poor prognosis in many cancers. Although telomerase activity has been found in chondrosarcomas, its prognostic significance in these malignant cartilage tumors is unknown. Malignancy in cartilage-derived tumors is assessed routinely by histomorphologic grading, but even well differentiated, low-grade lesions can metastasize. This unpredictable behavior greatly complicates the clinical treatment of cartilage tumors, making better prognostic indicators desirable. To address this issue we used immunohistochemistry to compare telomerase reverse transcriptase expression in a collection of 61 tumors consisting of malignant chondrosarcomas of varying grade and benign enchondromas. Associated case histories were reviewed to test the hypothesis that telomerase reverse transcriptase expression levels correlated with subsequent tumor recurrence. We found that the relative abundance of telomerase reverse transcriptase-expressing cells correlated significantly with grade and recurrence. These findings indicate that telomerase reverse transcriptase immunostaining may be a useful adjunct to the conventional three level grading system. PMID- 15346062 TI - Use of a smooth press-fit stem preserves physeal growth after tumor resection. AB - We conducted this study to determine whether the small-diameter, press-fit stem of a novel, noninvasive expandable endoprosthetic device implanted in the limbs of 6 pediatric patients with Stage IIB osteosarcoma affected the growth of the physis through which the stem was inserted. Local control of the tumor necessitated resection of the involved growth plate, and implantation of the device required penetration of the adjacent uninvolved bone, through the growth plate, by the stem of the device. We measured longitudinal growth and the rate of growth of the adjacent uninvolved bone in the salvaged limb and of the equivalent bone in the unoperated contralateral limb. In all cases but one (in which the patient's growth in the contralateral limb appeared complete), longitudinal growth continued in the limb into which the device was implanted: the adjacent uninvolved bone in the salvaged limb grew by an average of 2.4 cm, and the equivalent bone in the unoperated contralateral limb grew by an average of 2.3 cm. We conclude that implantation of a smooth, press-fit stem through the central portion of the uninvolved adjacent physis does not result in growth retardation or arrest. PMID- 15346063 TI - Lymph node metastasis in soft tissue sarcoma in an extremity. AB - For patients with soft tissue sarcoma in an extremity, the outcome is thought to be poor if lymph node metastasis develops. The purpose of this study was to examine the impact of lymphatic involvement from soft tissue sarcoma on patient survival. Thirty-nine (3.7%) of 1066 patients who had surgery for soft tissue sarcoma in an extremity had lymph node metastases develop. Three (20%) of 15 patients with epithelioid sarcoma, four (19%) of 21 patients with rhabdomyosarcoma, two (11.1%) of 18 patients with clear cell sarcoma, and two (11.1%) of 18 patients with angiosarcoma had lymphatic involvement. Thirty patients who had resection of involved lymph nodes had an estimated 5-year survival of 57%, whereas nine patients treated without surgery all died within 30 months. An estimated 4-year survival of 71% for patients with isolated lymph node metastases was significantly better than 21% for patients with synchronous systemic and lymph node involvement. There was no difference in outcome for patients with isolated lymphatic involvement compared with patients with American Joint Committee on Cancer Stage III extremity sarcomas. These results suggest that long-term survival is possible after surgical resection of lymphatic metastases from soft tissue sarcoma. The American Joint Committee on Cancer should consider separating isolated nodal metastases from systemic involvement in patients with Stage IV sarcoma. PMID- 15346064 TI - Dorsal transscaphoid-transtriquetral perilunate dislocation in pseudarthrosis of the scaphoid. AB - A patient reported that he sustained a minor fall on the outstretched hand in hyperextension, pronation, and in ulnar deviation. Initial radiographs suggested dorsal transscaphoid-transtriquetral perilunate dislocation. Traditionally, however, this injury is the result of a high-energy impact. A CT scan obtained after closed reduction of the dislocation revealed not only a fresh fracture of the triquetrum but also two corticalized fragments of the scaphoid. A former major fall on this hand and a normal scaphoid of the other hand made pseudarthrosis more likely than scaphoid bipartition. Arthrography revealed intact lunotriquetral and scapholunate ligaments, precluding the possibility of preexisting ligamentous instability. Pseudarthrosis of the scaphoid with a loss of scaphoid function as a mechanical tie-rod of the carpus is most likely responsible for this complex injury. This is the first clinical study that shows that single scaphoid discontinuity without preexisting ligamentous carpal instability may lead to complex perilunar dislocation in minor trauma. PMID- 15346065 TI - Does obesity affect the results of lumbar decompressive spinal surgery in the elderly? AB - The prevalence of obesity among the population is increasing, including in many elderly people. The purpose of this study was to evaluate whether lumbar spinal surgery in elderly patients with different body mass indices influences pain, satisfaction rate, and activities of daily living. Two hundred ninety-eight elderly patients (older than 65 years), 153 women and 145 men, who had decompressive laminectomy, discectomy, or combinations of these procedures during 1990 to 2000 were followed up. Indications for surgery included limitation in doing activities of daily living, severe pain, or both. The patients were classified into one of four categories in terms of their body mass index. The operative parameters, pain reduction, satisfaction rate, and activities of daily living using the Barthel index were assessed. The more obese patients were younger, tended to be female, and were more symptomatic. All four groups of patients had reduction in pain, improvement in activities of daily living, and were satisfied with the operation. Our data suggest that it is reasonable to operate on patients who are elderly and obese and who have lumbar symptoms, with the appropriate indications. PMID- 15346066 TI - Acetabular labral tears rarely occur in the absence of bony abnormalities. AB - We evaluated the percentage of patients with acetabular labral tears who have a structural hip abnormality detectable by conventional radiography. Records from our institution from 1996 through 2002 were reviewed to identify all patients with labral tears. Patients were excluded who had classic hip dysplasia, advanced osteoarthritis, or a history of pelvic or femoral osteotomy. The hip radiographs were evaluated for abnormalities of Tonnis angle, center-edge angle of Wiberg, acetabular version, femoral neck-shaft angle, congruency between the femoral head and acetabulum, anterior femoral head-neck offset, and presence of femoral head osteophytes. Twenty-seven of the 31 patients (87%) had at least one abnormal finding and 35% had more than one abnormality. Ten patients had a retroverted acetabulum, 16 had coxa valga, 11 had an abnormal femoral head-neck offset, and 14 had osteophytes on the femoral head. Four of 31 patients (13%) had no identifiable structural abnormalities. To our knowledge, this is the first study to document that the majority of patients with labral tears have a structural hip abnormality detectable with conventional radiographs. Familiarity with these structural abnormalities is important for early detection and accurate diagnosis, and may impact optimal treatment planning and prognosis. PMID- 15346067 TI - Twenty-five years followup of patients who had valgus osteotomy for arthritic hips. AB - We reviewed the long-term outcomes of intertrochanteric valgus femoral osteotomies in patients with arthritic hips to clarify any influencing factors. One hundred six patients (127 hips) were followed up during an average of 25 years. The average age of the patients at surgery was 42 years. The preoperative extent of degenerative change was classified radiologically into one of four grades according to the criteria of Tonnis. Radiographic measurements of acetabular coverage were made using AP radiographs obtained immediately after surgery. Thirty-eight patients (41 hips) had total hip arthroplasties; the 25 year survival rate was 69%. Radiologic evaluations of patients with mild preoperative degenerative changes (Tonnis Grade 1) improved and good clinical outcomes were obtained. In addition, radiologic evaluations of patients whose hips had better acetabular coverage (center-edge angle > 0 degrees, sharp angle < 50 degrees, or acetabular head index > 60%) also improved. However, radiographic measurements did not influence clinical scores. The mean score of patients younger than 50 years at surgery was higher than patients older than 50 years. The mean score of patients with unilateral hip involvement was higher than patients with bilateral involvement. Therefore, valgus osteotomies seem appropriate for younger patients with unilateral involvement. PMID- 15346068 TI - Reduced variability of acetabular cup positioning with use of an imageless navigation system. AB - Positioning the acetabular component is one of the most important steps in total hip arthroplasty; malpositioned components can result in dislocations, impingement, limited range of motion, and increased polyethylene wear. Conventional surgery makes use of specialized alignment guides provided by the manufacturers of the implants. The use of mechanical guides has been shown to result in large variations of cup inclination and version. We investigated acetabular cup alignment with the nonimage-based hip navigation system compared with a conventional mechanically guided procedure in 12 human cadavers. Postoperative cup position relative to the pelvic reference plane was assessed in both groups with the use of a three-dimensional digitizing arm. In the navigated group, a median inclination of 45.5 degrees and a median anteversion of 21.9 degrees (goals, 45 degrees and 20 degrees) were reached. In the control group, the median inclination was 41.8 degrees and the median anteversion was 24.6 degrees. The ninetieth percentile showed a much wider range for the control group (36.1 degrees-51.8 degrees inclination, 15 degrees-33.5 degrees anteversion) than for the navigated group (43.9 degrees-48.2 degrees inclination, 18.3 degrees-25.4 degrees anteversion). This cadaver study shows that computer-assisted cup positioning using a nonimage-based hip navigation system allowed for more consistent placement of the acetabular component. PMID- 15346069 TI - Mini-incision anterior approach does not increase dislocation rate: a study of 1037 total hip replacements. AB - Correct positioning of the prosthetic components in total hip replacements is important to prevent dislocations. Correct positioning is made easier by extensive approaches, but it also is possible using the mini-incision approach. The mini-incision used to facilitate early rehabilitation should not produce a higher dislocation rate than that of the more conventional approaches. The anterior surgical approach we describe allows for good exposure, despite the reduced size of the skin incision. Its length is 5-10 cm and usually 6-8 cm for patients with normal corpulence. Our mini-incision anterior approach using intermuscular planes allows a surgical approach to the hip and implantation of a total prosthesis with no muscle, tendon, or trochanteric section, even partially. This is not possible with any other surgical approach. A series of 1037 primary total hip replacements done between June 1993 and June 2000 was studied retrospectively. The dislocation rate was 0.96% (10 of 1037 hips). The mini incision approach allows for adequate positioning of the two prosthetic components. Preserving the muscular potential also may contribute to dynamic stabilization of the hip. PMID- 15346070 TI - Computer-assisted cup placement techniques in total hip arthroplasty improve accuracy of placement. AB - Malposition of the acetabular component during hip arthroplasty increases the occurrence of impingement, reduces range of motion, and increases the risk of dislocation and long-term wear. To prevent malpositioned hip implants, an increasing number of computer-assisted orthopaedic systems have been described, but their accuracy is not well established. The purpose of this study was to determine the reproducibility and accuracy of conventional versus computer assisted techniques for positioning the acetabular component in total hip arthroplasty. Using a lateral approach, 150 cups were placed by 10 surgeons in 10 identical plastic pelvis models (freehand, with a mechanical guide, using computer assistance). Conditions for cup implantations were made to mimic the operating room situation. Preoperative planning was done from a computed tomography scan. The accuracy of cup abduction and anteversion was assessed with an electromagnetic system. Freehand placement revealed a mean accuracy of cup anteversion and abduction of 10 degrees and 3.5 degrees, respectively (maximum error, 35 degrees). With the cup positioner, these angles measured 8 degrees and 4 degrees (maximum error, 29.8 degrees), respectively, and using computer assistance, 1.5 degrees and 2.5 degrees degrees (maximum error, 8 degrees), respectively. Computer-assisted cup placement was an accurate and reproducible technique for total hip arthroplasty. It was more accurate than traditional methods of cup positioning. PMID- 15346071 TI - Navigation improves accuracy of rotational alignment in total knee arthroplasty. AB - Successful total knee arthroplasty is dependent on the correct alignment of implanted prostheses. Major clinical problems can be related to poor femoral component positioning, including sagittal plane and rotational malalignment. A prospective randomized study was designed to test whether an optical navigation system for total knee arthroplasty achieved greater implantation precision than a nonnavigated technique. The primary variable was rotation of the femoral component in the transverse plane, measured from postoperative radiographs and computed tomography images. Sixty-four patients were included in the study. All patients received the Duracon total knee prosthesis. The patients were randomly divided into two groups: Group C patients had conventional total knee arthroplasty without navigation; Group N patients had total knee arthroplasty using a computer-assisted knee navigation system. Analysis showed that patients in Group N had significantly better rotational alignment and flexion angle of the femoral component than patients in Group C. In addition, superior postoperative alignment of the mechanical axis, posterior tibial slope, and rotational alignment was achieved for patients in Group N. The use of a navigation system provides improved alignment accuracy, and can help to avoid femoral malrotation and errors in axial alignment. PMID- 15346072 TI - Implant design affects knee arthroplasty kinematics during stair-stepping. AB - Knee implant motions have a direct influence on patient function and implant longevity. The purpose of this study was to determine if there were consistent differences in knee motions among three groups of knee implants. Two hundred thirteen knees in 173 patients, with 25 implant designs, were studied using fluoroscopy during stair-stepping. All knee implants were assigned to one of three groups based on the design: fixed-bearing posterior-stabilized, fixed bearing posterior cruciate-retaining, and mobile-bearing. All types of implants had the same pattern of internal/external rotations, but different designs had different anteroposterior translations. Seventy-five percent of posterior stabilized knee implants had a medial center of rotation, indicating posterior femoral translation with flexion. Sixty-three percent of cruciate-retaining fixed bearing knee implants had a lateral center of rotation. Eighty-six percent of mobile-bearing knee implants had a lateral center of rotation, indicating anterior femoral translation with flexion. Knee motion in patients with successful total knee arthroplasties is related directly to the constraints of the implant design. PMID- 15346073 TI - Effect of tibial slope or posterior cruciate ligament release on knee kinematics. AB - An experimental study using fresh human cadaver knees was designed to evaluate the effect of partial posterior cruciate ligament release or posterior tibial slope on knee kinematics after total knee arthroplasty. Varus and valgus laxity, rotational laxity, anteroposterior laxity, femoral rollback, and maximum flexion angle were evaluated in a normal knee, an ideal total knee arthroplasty, and a total knee arthroplasty in which the ligaments were made to be too tight in flexion. The total knee arthroplasty specimens then were subjected to either partial posterior cruciate ligament release or increased posterior tibial slope, and the tests were repeated. Posterior tibial slope increased varus and valgus laxity, anteroposterior laxity, and rotational laxity in the knee that had flexion tightness. Posterior cruciate ligament release corrected only anteroposterior tightness, and had no effect on the abnormal collateral ligament tightness. Increased posterior tibial slope significantly improved varus and valgus laxity and rotational laxity in the knee that was tight in flexion more than with release of the posterior cruciate ligament. Therefore increasing posterior tibial slope is preferable for a knee that is tight in flexion during total knee arthroplasty. PMID- 15346074 TI - Long-term results on abduction and external rotation of the shoulder after latissimus dorsi transfer for sequelae of obstetric palsy. AB - Transfer of the latissimus dorsi to the rotator cuff is widely used for restoring shoulder abduction and external rotation in sequelae of brachial plexus palsy; however, its long-term results are not well known. Because persistence of clinical benefits is crucial for children with brachial plexus palsy, the aim of our study was to evaluate retrospectively the long-term results of this transfer in 203 children. Patients were classified according to type of paralysis (C5-C6, C5-C6-C7, and complete), degree of preoperative shoulder function according to Gilbert, and age at surgery. Active abduction and external rotation were measured at 1, 3, 6, 10, and 15 years and the results were analyzed statistically. Children with sequelae of C5-C6 palsy gained in abduction and external rotation more than children with C5-C6-C7 or complete palsy. Patients with mild preoperative shoulder dysfunction achieved the best results. The data showed the clinical results were related to the type of paralysis and to preoperative shoulder function, but not to age at surgery. They also showed progressive deterioration of abduction began at 6 years despite preserved active external rotation. PMID- 15346075 TI - Forearm supination deformity after obstetric paralysis. AB - We retrospectively evaluated 66 patients who had radius osteotomies (44 patients; average age, 79.9 months) or soft tissue procedures (22 patients; average age, 57.4 months) to improve persistent supination at rest secondary to obstetric brachial plexus lesions and select the optimal treatment indication for each patient. Followups averaged 64.3 months and 72.57 months, respectively. In the osteotomy group there was a significant difference when preoperative supination at rest (average 31 degrees), intraoperative pronation obtained (average 114 degrees), and position of the forearm at last followup (average 92 degrees), were compared, whereas in the soft tissue procedures group there was not a significant difference between intraoperative pronation obtained (average 116 degrees) and forearm position at rest at last followup (average 108 degrees). Shoulder function was not affected by any of these interventions. Elbow extension was decreased at the last followup in both groups. Wrist and finger extension were improved in both groups. Complications included two delayed unions, one nonunion, and nine recurrences, all secondary to radius osteotomies. The procedure of choice for each patient must be based on preoperative forearm motion, condition of the proximal and distal radioulnar joints, triceps function, deformities of the shoulder and elbow, and hand function. PMID- 15346076 TI - Atypical slipped capital femoral epiphysis after radiotherapy and chemotherapy. AB - Atypical slipped capital femoral epiphysis after radiotherapy and chemotherapy is uncommon. Only 32 cases have been reported in the literature. Because patients may have slippage at atypical ages, we report two cases of slipped capital femoral epiphysis in children and review the 32 cases previously reported to heighten clinicians' awareness of this condition in patients who have received radiation and chemotherapy for pelvic tumors. The controversy over prophylactic pinning of the uninvolved hip in radiotherapy-associated slipped capital femoral epiphysis is unresolved. It may be justifiable to fix the nonslipped epiphysis if possible prodromal signs of abnormal radiographic findings are detected. Because radiotherapy and chemotherapy were used in the two children reported, it is not possible to state whether one or both forms of treatment were responsible for the atypical slipped capital femoral epiphysis. PMID- 15346077 TI - New indications for computer-assisted surgery: tumor resection in the pelvis. AB - The resection of recurrent malignant pelvic tumors was supported by a commercially available navigation system in three patients. Preoperatively three dimensional images from the pelvis were obtained by computed tomography or magnetic resonance imaging to identify the tumor extension. During surgery navigated tools oriented the surgeon to excise the tumor with adequate virtual margins. Navigation was helpful for tumor identification in one patient with a recurrent presacral mesenchymal chondrosarcoma. In the other two patients the tumor resection in the bone was done with three-dimensional observation of the osteotomies in the sacrum. In all three patients the histopathologic analysis confirmed that the neoplasms were excised accurately within their margins. We think that computer-assisted surgery is a potential method to increase the accuracy of tumor resections. PMID- 15346078 TI - Low-voltage electrochemotherapy with low-dose methotrexate enhances survival in mice with osteosarcoma. AB - Methotrexate plays a key role in adjuvant chemotherapy for treatment of osteosarcoma, but is used at a high dose because it can pass through a cell membrane only with difficulty. Therefore, if the drug delivery of methotrexate to the tumor could be enhanced, antitumor effect and survival would improve. We examined whether enhancement of the antitumor effect of electrochemotherapy was feasible by using low-dose methotrexate in mice with osteosarcoma. The tumor bearing mice were divided into four groups: no treatment, methotrexate treatment alone, electroporation alone, and methotrexate treatment followed by electroporation. In single-treatment series, the size of the tumors in mice treated with electrochemotherapy decreased substantially 6 days after treatment, whereas continuous growth was observed in the other groups. In the series of treatments repeated three times at 6-day intervals, the original tumors in the electrochemotherapy group decreased consistently and the tumors disappeared in four of seven animals within 16 days. In the other groups, the tumors continued to grow and all host animals died within 58 days. These results show the usefulness of electroporation to enhance the effects of low-dose methotrexate and the potential benefits of electrochemotherapy for the treatment of human osteosarcoma. PMID- 15346079 TI - Scapular chondrosarcomas have high rates of local recurrence and metastasis. AB - This is the first report of a large series of patients with scapular chondrosarcomas. The grade distributions, locations of the tumors in the scapula, surgical techniques, status of margins, chondrosarcoma subtypes, Enneking stages, adjuvant therapies, local recurrence rates, metastasis rates, and survival prognoses of patients with scapular chondrosarcoma were evaluated. Forty-seven patients treated between 1921 and 1999 were analyzed retrospectively. Grade 3 disease was significantly associated with a poorer survival prognosis when compared with Grades 1 or 2 disease. Patients with tumors smaller than 5 cm were treated mainly with partial scapulectomy, and patients with tumors larger than 5 cm often were treated with total scapulectomy. The survival prognoses of patients with intralesional resections at initial surgery showed a tendency toward poorer survival when compared with patients with wide resections at initial surgery. Metastasis and local recurrence (21.3% and 40.4%) were higher in scapular chondrosarcomas than rates reported for patients with general chondrosarcomas, and local recurrence or metastasis was associated with limited survival. The 5- and 15-year survival probabilities subsequent to diagnosis were 79% and 53%, respectively. The high rates of local recurrence and metastasis likely were caused by the difficult anatomic relationships encountered during scapular resections. This study shows the importance of wide margins which must be achieved to provide local disease control. PMID- 15346080 TI - Segmental amputation for recurrent malignant bone tumors of the distal femur. AB - We describe an alternative method for lengthening a short femoral stump after wide amputation of a malignant bone tumor of the distal femur in two patients. The method consists of two procedures during the operation. The first procedure is amputation of the affected cylindrical segment of the involved limb. The second procedure is elongation of the amputation stump using the tumor-free segment of the ipsilateral lower leg as a free composite osseous myocutaneous graft. Both patients had good function and were satisfied with the results with no complications or tumor recurrence 4 years postoperatively. We think the segmental amputation is a good procedure that results in a longer functional stump in patients who have above-knee amputation. PMID- 15346081 TI - Advantages of submuscular bridge plating for complex pediatric femur fractures. AB - Conventional treatments of pediatric femoral shaft fractures may result in an unacceptable rate of complications, especially in complex fractures. These fractures include high-energy injuries resulting in unstable fracture patterns, fractures in the proximal or distal third, and fractures occurring in large or multiply injured children. Our goal was to evaluate whether a minimally invasive submuscular bridge plating technique provides stability for early functional treatment (without protective casting or bracing) and predictable healing. Fifty one patients with an average age of 10 years were studied. Sixty-seven percent had high-energy injuries and 55% had unstable fracture patterns. With an average followup of 14.2 months, all fractures united with excellent clinical results. Two (4%) significant complications occurred: fracture of one 3.5-mm LC-DCP Ti plate, and refracture of a pathologic fracture after early plate removal. Four patients (8%) had a leg-length discrepancy ranging from 23-mm short to 10-mm long. The average operative time was 106 minutes, with average fluoroscopy time of 84 seconds. Procedures were done by 15 surgeons in five university medical centers. This technique offers the advantage of adequate stability for early functional treatment and predictable healing with maintenance of length and alignment for all pediatric femoral shaft fractures. PMID- 15346082 TI - Femur-LISS and distal femoral nail for fixation of distal femoral fractures: are there differences in outcome and complications? AB - We evaluated the functional and radiologic outcomes after stabilization of distal femoral fractures using the distal femoral nail and a less invasive stabilization system to determine if the new implants are superior to other implants (especially the condylar blade plate) regarding the rates of axial deviation, nonunion, and infection and if one of these new implants (Less Invasive Stabilization System, or distal femoral nail) is superior to the other. Two groups, each with 16 patients, were documented prospectively and the results were compared. To record the findings objectively, the Lysholm-Gillquist score was used. A conversion procedure was done in two patients in the plate group and one patient of the nail group. At the 1-year followup mobility of the knee was on average 110 degrees in the plate group and 103 degrees in the nail group. The Lysholm-Gillquist score did not show any significant differences between the groups. There were clinically relevant varus or outer rotation deviations in three patients in the plate group and two patients in the nail group. The two minimally invasive implants used were good in terms of technique and outcome for treatment of distal femoral fractures and did not differ significantly for epidemiology, fracture type, conversion procedures, infection rate, malalignments, and subjective and objective findings at the 1-year followup. They were also superior to the condylar plate in terms of infection and axial malalignments. PMID- 15346083 TI - The tension required at repair to reappose the supraspinatus tendon to bone rapidly increases after injury. AB - Rotator cuff tears occur frequently and can cause significant pain and reduced shoulder function. A high percentage of patients are satisfied after surgical repair of rotator cuff tears, but a smaller percentage of patients with chronic tears continue to have pain and poor shoulder function. This may be partly attributable to an increase in the repair tension, the force required at repair to reappose the tendon to its original insertion site on the humerus. Increases in repair tension have been shown to occur for long-standing ruptures of the supraspinatus tendon, but the precise tension at various times after injury are unknown. Therefore, the objective of the current study was to determine the repair tension at various times after a rotator cuff tear. This was achieved by creating a full-thickness supraspinatus tendon tear in a rat model and measuring the mechanical characteristics of the musculotendinous unit at 0, 2, 4, 9, and 16 weeks after injury. The repair tension rapidly increased initially after injury followed by a progressive, but less dramatic, increase with additional time. These findings suggest that rotator cuff tears should be repaired early in the clinical setting. Future studies will investigate the effect of repair tension on tendon to bone healing after repair. PMID- 15346084 TI - Cartilage tolerates single impact loads of as much as half the joint fracture threshold. AB - We hypothesized that one mechanical insult could affect cellular proliferation, matrix turnover, and the structural integrity of cartilage, and that these effects would be dose dependent and time dependent. One impact load of low impact (14.4 MPa +/- 2.1 MPa), medium impact (22.8 MPa +/- 5.8 MPa), or high impact (55.5 MPa +/- 12.6 MPa) was administered to the posterior aspect of the medial femoral condyle of New Zealand White rabbits using a previously validated pendulum device. Animals were euthanized at 2, 6, and 12 weeks after impact, and the impacted and sham (contralateral limb) cartilage were harvested. Each specimen was assessed by light microscopy and by immunohistochemical methods. Although impacted specimens had greater loss of proteoglycan staining than sham cartilage, these changes were neither dose dependent nor time dependent. No structural damage, enzymatic proteoglycan or collagen breakdown, or cellular proliferation was identified in the different impact groups. Articular cartilage is a resilient tissue, particularly in situ, and can tolerate single impact loads of as much as 45% of the joint fracture threshold without considerable disruption or degradation. PMID- 15346085 TI - Thigh mass in a 6-year-old girl. PMID- 15346086 TI - Approaches to restenosis: mechanical and pharmacological strategies. Overview. PMID- 15346087 TI - Prevention of restenosis after stenting: the emerging role of inflammation. AB - Restenosis after stent implantation is mainly caused by neointimal proliferation through the stent struts. Experimental studies indicate a marked activation of inflammatory cells at the site of stent struts, which are likely to play a key role in the process of neointimal proliferation and restenosis. Coronary stenting is a strong inflammatory stimulus and the acute systemic inflammatory response to local inflammation produced by coronary stenting is highly individual and predicts restenosis and event-free survival. The benefit of anti-inflammatory therapy during the periprocedural period and long-term follow-up is dependent on the inflammatory status. Measurement of cytokine and acute phase proteins, such as C-reactive protein, may be important to identify high-risk subjects and develop specific treatment tailored to the individual patient. PMID- 15346088 TI - Drug-eluting stents: caution and concerns for long-term outcome. AB - Recent publications on drug-eluting stents (DES) report a significant reduction in restenosis rates as compared to bare metal stents in patients mostly with single vessel disease. We have recently observed however, late stent thrombosis following CYPHER DES implantation. The patient developed a hypersensitivity reaction around stent struts limited to the polymer with aneurysmal dilatation and extensive inflammation of the arterial wall in the absence of vascular healing. This incidence promotes a cautionary view and perhaps supports the use of DES only in high-risk patients. PMID- 15346089 TI - The role of brachytherapy and Cutting Balloon angioplasty in the current treatment of stent restenosis. AB - The Gamma I, START and INHIBIT trials conclusively demonstrate the feasibility, safety and efficacy of intracoronary radiation as the treatment of choice for stent restenosis. Further reports confirm this finding and extend the indications. Vascular brachytherapy should be made available for all patients with diffuse stent restenosis. Specific devices such as cutting balloons may improve the procedure but does not seem to have an impact alone on the evolution. PMID- 15346090 TI - The role of platelet inhibition in the drug-eluting stent era. AB - The use of antiplatelet agents in the management of patients undergoing percutaneous coronary interventions involves the administration of aspirin, thienopyridines, and in high-risk patients, GPIIb-IIIa antagonists. Drug-eluting stents now account for greater than 50% of stenting procedures. This review focuses on the limited available data describing the use of antiplatelet agents in patients undergoing drug-eluting stent implantation. PMID- 15346091 TI - Association between C-reactive protein and hypertension in healthy middle-aged men and women. AB - OBJECTIVE: To ascertain whether C-reactive protein (CRP), an inflammatory marker related to increased cardiovascular risk, is associated with blood pressure in a sample of healthy, middle-aged people. METHODS AND RESULTS: A case-control study among 904 participants, 39-50 years old, from a cardiovascular risk screening study. Participants with systolic blood pressure > or =140 mmHg or diastolic blood pressure > or =90 mmHg (n=120) were considered as case participants and all others as control participants (n=784). Exposure was defined using quintiles of high-sensitivity CRP among control participants. A continuous increase in blood pressure was observed across CRP quintiles. Systolic blood pressure increased 1.17 mmHg [95% confidence interval (CI), 0.60-1.74] and diastolic blood pressure 1.04 mmHg (95% CI, 0.64-1.45) from one quintile to the next. The prevalence of hypertension was 13.3% and it increased with CRP exposure: Q1, 8.9%; Q2, 11.9%; Q3, 12.2%; Q4, 14.3%; and Q5, 18.6%. After adjustment for sex, obesity, race, serum insulin level and family history of coronary heart disease, odds ratios for hypertension increased progressively across CRP quintiles. Participants in the highest CRP quintile were 2.35 times more likely to have hypertension than those in the lowest quintile (P=0.03, trend test P=0.04). CONCLUSION: These results are consistent with a continuous, independent association between serum CRP and elevated blood pressure. PMID- 15346092 TI - Correlation of changes in nitric oxide synthase, superoxide dismutase and nitrotyrosine with endothelial regeneration and neointimal hyperplasia in the balloon-injured rabbit subclavian artery. AB - OBJECTIVES: Alterations in nitric oxide (NO) and superoxide production within the wall of injured vessels may modulate the development and eventual extent of neointima after balloon injury. METHODS: In this study we have characterized a rabbit model of subclavian artery injury and have used immunocytochemistry to detect NO synthase (NOS) isoforms, Cu-Zn superoxide dismutase (SOD) and nitrotyrosine in the injured vessel from 2 h to 28 days after injury. RESULTS: At 48 h after injury, when cellular proliferation that will ultimately form the neointima is commencing, there was upregulation of inducible NOS, Cu-Zn SOD and nitrotyrosine. Recovery of endothelial NOS occurred at 28 days after injury, when the neointima is stabilizing and cellular proliferation has slowed down. There was no increase in neuronal NOS at any time point studied. CONCLUSIONS: NO may serve to limit the development of neointima while superoxide may attenuate the effect of NO by formation of peroxynitrite, detected as increased nitrotyrosine staining. Upregulation of Cu-Zn SOD would limit superoxide both at sites of inflammation in the vessel wall from 48 h and in the adventitia up to 28 days after injury. Very early intervention to protect NO may reduce neointimal size. PMID- 15346093 TI - The effect of long-term clopidogrel use on neointimal formation after percutaneous coronary intervention. AB - OBJECTIVE: The purpose of this study was to evaluate the long term effect of clopidogrel-based antiplatelet therapy on neointimal formation. METHODS: This study comprised 78 patients with typical stable angina pectoris or documented myocardial ischaemia, and with only one angiographic lesion in one native coronary artery undergoing successful stent implantation without predilatation with C-reactive protein levels < or =5 mg/l at 72 h after the procedure. All patients received dual antiplatelet therapy with 75 mg/day clopidogrel and 300 mg/day aspirin for four weeks. Clopidogrel was switched to isochronous placebo in half of the patients (n=39) at the end of the fourth week. This allocation was maintained for 20 weeks, and at week 24 of the study, coronary angiography and intravascular ultrasound imaging were performed again in all cases in order to evaluate the changes that had occurred in the in-stent neointimal formation; rates of restenosis were also recorded RESULTS: At the end of the follow-up period, angiographic stenosis diameter and restenosis rates were smaller in the clopidogrel group than in the placebo group (23.3% versus 35.6%, p=0.05 and 5.12% versus 10.25%; p=0.03 respectively); the intravascular ultrasonographic neointimal cross sectional area was also smaller in the clopidogrel group (3.6 +/ 2.7 mm(2) versus 5.2 +/- 2.5 mm(2), p=0.03). CONCLUSIONS: Long-term clopidogrel administration significantly reduced neointimal formation at the stent site as well as reducing major clinical events in patients who did not develop high-risk systemic inflammatory response after percutaneous coronary intervention. PMID- 15346094 TI - Diabetes mellitus and unstable coronary artery disease: improved clinical outcome of coronary artery stenting in an era of glycoprotein IIb/IIIa inhibitors and lipid-lowering therapy. AB - OBJECTIVE: The short- and long-term clinical outcomes of coronary artery stenting in diabetic patients with unstable coronary artery disease were assessed and compared with a cohort of non-diabetic patients in the context of platelet glycoprotein IIb/IIIa inhibitors and lipid-lowering therapy. METHODS: The study population comprised 252 consecutive patients with unstable angina who underwent coronary artery stenting; of these 46 were diabetic and 206 were non-diabetic. In hospital results and clinical outcome during follow-up (24 +/- 13 months, range 7 56 months) were obtained in all patients without major in-hospital complications. Survival curves and multivariate models for any late clinical event were reported. RESULTS: A high clinical success rate and no difference in in-hospital complications between the two groups of patients were observed. Complete revascularization rate was similar in diabetic and non-diabetic patients (48% compared with 52%). A greater proportion of diabetic than non-diabetic patients received IIb/IIIa inhibitors during the procedure and lipid-lowering drugs at hospital discharge (87% compared with 46%, P=0.001 and 83% compared with 61%, P=0.006 respectively). At 2-year clinical follow-up, the incidences of death and myocardial infarction were similar in both groups; the need for any revascularization was only slightly higher in diabetic patients (P=NS). Incomplete revascularization and multi-vessel disease were independent predictors of any revascularization. CONCLUSIONS: Coronary artery stenting combined with glycoprotein IIb/IIIa inhibitor infusion and long-term lipid-lowering therapy is an effective therapeutic strategy in diabetic patients with unstable coronary artery disease and is associated with good short- and long-term results, comparable to those observed in non-diabetic patients. PMID- 15346095 TI - Impact of invasive strategy for the management of patients with cardiogenic shock after acute myocardial infarction. AB - OBJECTIVE: This study evaluates the influence of early revascularization (with percutaneous transluminal coronary angioplasty (PTCA) and coronary surgery) on short- and long-term survival in patients with cardiogenic shock complicating acute myocardial infarction (AMI). METHODS AND RESULTS: In-hospital and 6-month survival were retrospectively determined on day 193 (65-270, median +/- 25th and 75th percentiles) in 87 patients who either underwent early invasive reperfusion (group A, n=60) or those who were treated conservatively (group B, n=27). In hospital mortality was 37% in group A and 56% in group B (P=0.192). Six-month mortality was statistically lower in group A than in group B (30 patients (50%) compared with 25 patients (93%), P=0.005). Being a woman and older age were found to be factors increasing mortality. Lower mortality in the long term was strongly associated with revascularization (odds ratio=0.08, 95% confidence interval=1.54 109). PTCA was found to be an independent predictor of long-term survival (odds ratio= 0.22, 95% confidence interval=0.049-1.00, P=0.050), by multiple logistic regression. CONCLUSIONS: In conclusion, this study suggests that early revascularization improves long-term survival of patients with cardiogenic shock complicating AMI, even after adjustment for baseline differences between patients who underwent early revascularization and those who did not. PMID- 15346096 TI - Bibliography. Current world literature. PMID- 15346097 TI - Improving the pharmacological regimen for patients with diabetes mellitus. AB - Type 2 diabetes mellitus affects nearly 17 million people in the United States, and prevalence rates are expected to double within 2 decades. Although there has been a downward trend in cardiovascular morbidity and mortality in recent years, cardiovascular disease remains the leading cause of death among patients with diabetes. This observation had led many to reevaluate current treatment goals and pharmacologic regimens for at-risk patients with type 2 diabetes mellitus. This review focuses on the current adjunctive pharmacologic treatment regimen that is well-suited for these patients. PMID- 15346098 TI - Circadian variations in cardiovascular disease: chronotherapeutic approaches to the management of hypertension. AB - Blood pressure (BP) exhibits strong circadian variation, and this variation may contribute to the increase of acute cardiovascular events that peak in the morning hours. Reducing morning BP may prevent these occurrences, so identifying data on the true duration of action of antihypertensive agents is essential. Ambulatory BP monitoring has uncovered important differences in commonly used once-daily therapies and has provided insights into the cardiovascular risks associated with BP variability. This article will explore chronotherapeutic antihypertensive agents that have been formulated to address the circadian challenges in controlling BP, and will consider the implications of chronotherapeutics in managing cardiovascular disease. PMID- 15346099 TI - Aspirin resistance: current concepts. AB - Aspirin is an effective antiplatelet agent with proven benefit in the prevention of atherothrombotic complications of cardiovascular disease. The antithrombotic effects of aspirin, however, are variable among individuals and this might explain, in part, why the absolute risk of recurrent vascular events in patients receiving aspirin therapy remains relatively high (8% - 18% after 2 years). Although formal diagnostic criteria are lacking, aspirin resistance generally describes the failure of aspirin to produce an expected biological response or the failure of aspirin to prevent atherothrombotic events. Aspirin resistance has been reported to occur in 5% to 45% of the general population; therefore, its detection is potentially of clinical importance. The biological mechanisms, population prevalence, laboratory methods for detection, and clinical relevance of aspirin resistance are discussed in this review. PMID- 15346101 TI - Cardiac mass in the setting of acute myocardial infarction. AB - When a middle-aged woman with a complex cardiac history presented with a cardiac mass in the setting of an evolving myocardial infarction, it created a diagnostic dilemma. This case report describes the diagnostic evaluation of such a patient and discusses the rare finding of an intramyocardial hematoma. PMID- 15346102 TI - HIV disease and an atherosclerotic ascending aortic aneurysm. AB - Cardiovascular dysfunction appears to be an important complication of human immunodeficiency virus (HIV) infection and is being reported with greater frequency. There have been recent reports in the literature of HIV patients who suffer from vascular lesions such as large artery vasculopathy secondary to vasculitis, as well as accelerated atherosclerosis of the coronary arteries. The latter has been linked to patients on protease inhibitors that are used as part of a highly active antiretroviral therapy (HAART) regimen and have also been implicated in a lipodystrophy syndrome. We report a rare case of an HIV-infected patient on HAART who presented with a large ascending aortic aneurysm associated with symptomatic severe aortic regurgitation. A noteworthy finding on pathological analysis of the aorta was an etiology of accelerated atherosclerosis rather than the more expected vasculitis. PMID- 15346103 TI - An update on diagnostic criteria for diabetes and the metabolic syndrome. PMID- 15346105 TI - Ranolazine: a new drug and a new paradigm for management of myocardial ischemia and angina. PMID- 15346104 TI - Conventional risk factors and cardiac events--debunking an old myth about prevalence. PMID- 15346106 TI - BMP and activin receptor expression in lens development. AB - PURPOSE: Members of the TGFbeta super-family have been shown to play important roles in lens development, including lens placode formation and fiber differentiation, and also induce changes characteristic of some forms of cataract. Previous studies demonstrated expression of TGFbeta receptors during lens morphogenesis. However, the expression patterns of activin and BMP receptors or their signaling mediators, the Smad proteins, have not been well documented. In this study we examine the spatio-temporal expression patterns of activin receptors (ActRIIA, ActRIIB, ALK1, and ALK2), BMP receptors (BMPRII, ALK3, and ALK6), and the distribution of the phosphorylated forms of Smad1 and Smad2 during normal lens development (E12-P21) and aberrant development in transgenic mice that express dominant negative TGFbeta receptors. METHODS: RT-PCR was used to identify receptor expression in total RNA isolated from P2 and P21 rat lenses. cDNAs were cloned and used for in situ hybridization analysis of spatio-temporal expression patterns in wild type and transgenic (OVE550 and OVE591) lenses. Expression of ALK3 was also examined by immunofluorescence and immunoblotting. Antibodies for phosphorylated forms of Smad1 and Smad2 were used to examine activation of BMP and activin signaling. RESULTS: RT-PCR of RNA from postnatal lenses showed distinct expression of ActRIIA, ActRIIB, BMPRII, and ALK3 but not ALK1, ALK2, or ALK 6. In situ hybridization with specific probes for BMPRII, ActRII, and ALK3 showed ubiquitous expression in ectoderm, lens pit, optic vesicle, and peri-optic mesenchyme during early lens formation at E12. During subsequent lens differentiation, from E14 onwards, expression of these receptors became increasingly restricted to the lens epithelium and to the equatorial region, including the germinative and transitional zones, where cells proliferate and commence differentiation, respectively. Expression for both receptors declined rapidly with fiber differentiation and maturation. Immunofluorescence with specific antibodies for phospho-Smad1 and phospho-Smad2 showed distinct localization of these signaling mediators in epithelial cells of the germinative zone and in fibers undergoing early differentiation in the transitional zone. Further investigation of the expression of these receptors in lenses of transgenic mice, which ectopically express a truncated TbetaRII, showed marked up regulation and aberrant expression of ALK3, but not BMPRII or ActRII. CONCLUSIONS: These results indicate that multiple members of the TGFbeta family have the potential to signal during lens fiber differentiation and suggest there may be cross-talk between different signaling pathways. PMID- 15346107 TI - Arthroscopic visualization of the posterior compartments of the knee. AB - PURPOSE: The purpose of this study is to test the hypothesis that the efficacy of routine arthroscopic evaluation of the posterior compartments of the knee outweighs the morbidity or inefficiency. TYPE OF STUDY: Prospective cohort analysis. METHODS: One hundred consecutive knees underwent posteromedial and posterolateral evaluation according to an algorithm designed to maximize efficiency of technique. Loose bodies were defined as expected when preoperative imaging or arthroscopy of the rest of the knee revealed loose bodies. The number of attempts required for visualization or visualization not accomplished, morbidity (subjective femoral condylar scuffing or complications associated with the technique), and findings were recorded. RESULTS: With regard to posteromedial, directly inserting the camera (as opposed to an obdurator) resulted in instrument breakage. In this study, 82% of posteromedial compartments were visualized on the first attempt, and 3% were not visualized. We found that 66% of femoral condyles had no damage, 28% mild, 3% moderate, and 3% severe. Loose bodies were found in 4 of 11 (36%) when expected and 0 of 86 when not expected, a significant difference. Five other posteromedial positive findings were observed. With regard to posterolateral, 93% of compartments were visualized on the first attempt, directly with the camera via the anterolateral portal. All were visualized. We found that 96% of femoral condyles had no damage, 4% mild, and 0% moderate or severe. Damage occurred significantly less frequently than with posteromedial. No findings were noted. CONCLUSIONS: Despite inefficiency, we recommend an obdurator for posteromedial visualization via the anterolateral portal. Posteromedial visualization was associated with morbidity but was efficacious in some cases and is strongly recommended when loose bodies are expected. Posterolateral visualization directly with the camera and via the anterolateral portal is efficient and associated with minimal morbidity but was not efficacious in the cohort evaluated. LEVEL OF EVIDENCE: Level II. PMID- 15346108 TI - Arthroscopic autogenous osteochondral transplantation for treating knee cartilage defects: a 2- to 5-year follow-up study. AB - PURPOSE: The purpose of this retrospective analysis was to evaluate the medium term results of arthroscopic treatment of chondral and osteochondral lesions of the knee with autogenous osteochondral transplantation (AOT). TYPE OF STUDY: Case series. METHODS: Thirty-three skeletally mature patients with symptomatic, full thickness cartilage lesions of the femoral condyles, between 1 and 2.5 cm in diameter, were treated with arthroscopic AOT. Grafts were harvested from the superior and lateral intercondylar notch and press-fit into holes drilled into the defect. All patients were evaluated both preoperatively and postoperatively with the Lysholm knee score, International Knee Documentation Committee (IKDC) Standard Evaluation Form, and knee joint radiographs. RESULTS: Thirty patients of a mean age of 44.6 years were followed-up for a mean time of 45.1 months. Symptom duration ranged from 1 month to 15 years (median, 9.5 months). The mean Lysholm score significantly improved from 43.6 preoperatively to 87.5 postoperatively ( P <.001). Excellent or good outcome was accomplished in 25 of the patients (83%). Using the IKDC assessment, 26 of the patients (87%) reported their knee as being normal or nearly normal. Repeat arthroscopy with needle biopsy of the graft was performed in 9 patients. Seven of these had complete healing and 2 partial healing. The histologic examination revealed viable chondrocytes and normal hyaline cartilage in the completely healed cases. Congruency of the articular surface was restored in 11 of 12 patients (92%) who underwent magnetic resonance imaging examination. Abnormal marrow signal in the subchondral bone beneath the region of cartilage repair was present in 9 of 12 patients (75%), even 4 years after the procedure. CONCLUSIONS: Arthroscopic AOT is an effective and safe method of treating symptomatic full-thickness chondral defects of the femoral condyles in appropriately selected cases. However, further studies with long-term follow-up are needed to determine if the grafted area will maintain structural and functional integrity over time. LEVEL OF EVIDENCE: Level IV, therapeutic, case series (no, or historical, control group). PMID- 15346109 TI - Influence of obesity on outcome after knee arthroscopy. AB - PURPOSE: To compare the clinical outcomes in overweight women with those of a group of normal-weight women 4 to 11 years after arthroscopic debridement of the knee. METHODS: A questionnaire containing the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), the short form health survey SF-36, and a survey regarding satisfaction with surgery was mailed to 121 patients. Questionnaire data were supplemented by chart review. RESULTS: Ninety patients (74%) returned completed questionnaires. All scored significantly lower than age matched normative data on all domains of the SF-36. The normal-weight group had better scores than the obese group, particularly in the physical functioning ( P =.01), role physical ( P =.05), mental health ( P =.11), and role emotional ( P =.11) domains. They were also more satisfied with their surgery ( P <.05), more likely to indicate that they would have the surgery again, and had fewer recurrences of knee pain ( P <.05). CONCLUSIONS: Overweight women have poorer outcomes related to quality of life and satisfaction with surgery than normal weight women. LEVEL OF EVIDENCE: Level III, Case control study. PMID- 15346110 TI - The value of clinical examination versus magnetic resonance imaging in the diagnosis of meniscal tears and anterior cruciate ligament rupture. AB - PURPOSE: The purpose of this study was to compare the accuracy of clinical examination versus magnetic resonance imaging (MRI) in diagnosing meniscal and anterior cruciate ligament (ACL) pathology. TYPE OF STUDY: Prospective, longitudinal study. METHODS: Between August 2001 and December 2001, we prospectively evaluated 50 consecutive patients (37 male, 13 female) with 65 pathologies of medial meniscal tears, lateral meniscal tears, and/or ACL rupture. The average preoperative period for the patients was 5 weeks (range, 5 days to 5 months) and their mean age was 22 years (range, 12 to 42 years). After initial clinical examination, the same sports medicine fellowship-trained orthopaedic surgeon (10-year practice profile of 100% sports medicine) evaluated the MRI of the patients and performed their arthroscopic procedure. Accuracy, sensitivity, specificity, and positive and negative predictive values were calculated comparing clinical examination, MRI, and arthroscopic evaluation. RESULTS: There was no statistical difference between MRI or clinical examination in diagnosing medial or lateral meniscal tears or ACL tears ( P >.05). The accuracy of the clinical examination and MRI evaluation was equal for diagnosing meniscal tears and ACL ruptures. CONCLUSIONS: A well-trained qualified surgeon can safely rely on clinical examination for diagnosing meniscal and ACL injuries. Clinical examination is at least as accurate as MRI in the skilled orthopaedic surgeon's hand. MRI should be reserved for more complicated and confusing cases. The routine ordering of an MRI scan of the knee before examination by a well-trained orthopaedic surgeon is not recommended. LEVEL OF EVIDENCE: Level II, diagnostic. PMID- 15346111 TI - Internal impingement: findings on magnetic resonance imaging and arthroscopic evaluation. AB - PURPOSE: The purpose of this study was to examine preoperative shoulder magnetic resonance imaging (MRI) and intraoperative arthroscopic findings in throwing athletes with a clinical diagnosis of internal impingement. TYPE OF STUDY: Retrospective review. METHODS: A retrospective review of the surgical indications for 769 shoulder arthroscopic procedures performed from 1997 to 2000 revealed 9 throwing athletes with a diagnosis of severe internal impingement. Each had failed treatment that included a minimum of 3 months of rehabilitation. Preoperative MRIs in all 9 were read by a fellowship-trained musculoskeletal radiologist blinded to the operative findings. Findings at arthroscopy were culled from the operative reports. RESULTS: On both MRI and at arthroscopy, all 9 athletes had posterosuperior labral lesions, 1 had a humeral head articular cartilage lesion, and 1 had a SLAP lesion. Additional findings on MRI included humeral head edema, subchondral cystic changes, posterosuperior glenoid sclerosis, and rotator cuff tendonopathy predominantly in the infraspinatus. At arthroscopy, there were additional findings of articular-sided partial rotator cuff tears not shown on MRI. CONCLUSIONS: Both MRI and arthroscopic findings described labral and humeral head articular cartilage lesions. These findings may be indicative of internal impingement in throwing athletes. LEVEL OF EVIDENCE: Level III, diagnostic. PMID- 15346112 TI - A serial comparison of arthroscopic repairs for partial- and full-thickness rotator cuff tears. AB - PURPOSE: To compare the outcome of arthroscopic rotator cuff repair and subacromial decompression in partial-thickness rotator cuff tears (PTRCT) with those in full-thickness rotator cuff tears (FTRCT). TYPE OF STUDY: Prospective serial follow-up study. METHODS: Of 46 consecutive patients who were treated with arthroscopic rotator cuff repair, 42 patients who were followed-up serially for 2 years were enrolled as study subjects. The average age of the patients at the time of the operation was 55 years, and the mean duration of follow-up was 34 months. The subjects included 22 cases of PTRCT and 20 cases of FTRCT. RESULTS: At the final follow-up, the PTRCT group showed changes in scores from 7.2 to 0.9 for average pain and from 34 to 91 for the shoulder functional evaluation score of the American Shoulder and Elbow Society (ASES score). The FTRCT group showed changes in scores from 7.6 to 1.2 for pain and from 29 to 88 for the ASES score. There were no significant differences between the 2 groups ( P >.05). The average range of shoulder motion was significantly improved in both groups at the final follow-up versus their preoperative values. Evaluation at the final follow-up showed that 93% of the total subjects showed good or excellent results, and 95% showed satisfactory results with regard to pain reduction and functional outcomes. The 2 fair results were the result of acromioclavicular arthritis. CONCLUSIONS: It may be anticipated that arthroscopic rotator cuff repair and subacromial decompression will give satisfactory postoperative outcomes in both PTRCT and FTRCT in terms of pain relief and functional recovery. However, careful preoperative examination of the acromioclavicular joint is critical to avoid procedural failure. LEVEL OF EVIDENCE: Level IV. PMID- 15346113 TI - Kim's lesion: an incomplete and concealed avulsion of the posteroinferior labrum in posterior or multidirectional posteroinferior instability of the shoulder. AB - PURPOSE: The purpose of this article is to report a new clinical entity of posterior instability of the shoulder and the results of its treatment. TYPE OF STUDY: Case series. METHODS: The Kim's lesion, which is an incomplete and concealed avulsion of the posteroinferior labrum, was arthroscopically identified in 15 patients who presented with posterior or multidirectional posteroinferior instability. Patients were treated by arthroscopic labroplasty and capsular shift. At a minimum follow-up of 2 years, the outcome was evaluated using subjective (pain and function visual analogue scale) and objective (UCLA, ASES, and Rowe scores) measurements. RESULTS: When visualized under an arthroscope, Kim's lesion apparently had an intact labral attachment and appeared to have a superficial crack at the junction between the articular cartilage of the glenoid and the posteroinferior labrum. However, probing of the lesion revealed detachment of the deep portion of the posteroinferior labrum. The posteroinferior labrum was flat with loss of normal height, which resulted in the retroversion of the chondrolabral glenoid. Incision of the superficial portion of the lesion exposed a loose deep portion of the labrum. Labroplasty was performed to restore the labral height, as well as capsular shift with or without rotator interval closure. The surgical outcome was satisfactory in 14 patients and unsatisfactory in 1 patient. Shoulders were stable in all patients with unidirectional posterior instability. There was 1 recurrence of multidirectional posteroinferior instability. CONCLUSIONS: Kim's lesion is an incomplete avulsion of the posteroinferior labrum, which is concealed by apparently intact superficial portion. The clinical significance of this lesion is the need for surgeons to convert this concealed incomplete lesion to a complete tear and repair it with the posterior band of the inferior glenohumeral ligament. A failure to address this lesion may result in persistent posterior instability. LEVEL OF EVIDENCE: Level IV, therapeutic, Case Series. PMID- 15346114 TI - Endoscopic carpal tunnel release: modification of Menon's technique and data from 191 cases. AB - PURPOSE: Endoscopic carpal tunnel release using Menon's technique has been shown to reduce recovery time, although previous studies have shown that there is still a considerable risk of nerve complications. The purpose of this study was to evaluate data from 227 hands of 191 patients who underwent releases with Menon's technique. TYPE OF STUDY: Prospective study. METHODS: Endoscopic releases were performed as an outpatient intervention by one surgeon. Follow-up evaluations included were analysis of satisfaction, quantitative measurements of grip strength, return to work time, and complications. After the surgery on the first 50 hands in 41 consecutive cases, the technique was modified. Prospectively, changes in the technique are due to difficulties maintaining the knife within the center of the cannula's slot and a relatively high complication rate. In the modified technique, a 2.7-mm 25 degrees endoscope and a triangular diamond-tipped knife were used to allow for more room for the instruments and a more safe procedure. RESULTS: During the first month after the surgery, 91% of the patients had better subjective satisfaction scores and by 12 weeks, 81% obtained 75% to 100% or greater grip strength. Within 3 weeks, 70% of the patients had returned to work. Twelve of 50 hands operated on using the original technique had nerve disturbance diagnosed at follow-up. Three of these 12 patients developed reflex sympathetic dystrophy. One patient had partial median nerve injury that was repaired at the time of the index operation. There was no serious complication observed in 177 hands of 150 cases operated on using the modified Menon's technique, except one postoperative hypoesthesia along the long and ring fingers that improved with time. CONCLUSIONS: The procedure is suitable for outpatient surgery and the risk of inadvertent damage to the neurovascular structures can be dramatically reduced with the modifications to the technique. LEVEL OF EVIDENCE: Level IV. PMID- 15346116 TI - The keeled acromion: an aggressive acromial variant--a series of 20 patients with associated rotator cuff tears. AB - This study describes our experience with an unusual type of acromial spur. In the last 7 years, we have treated 20 patients with a previously undescribed "keeled" acromion. The keel refers to a central, longitudinal, downward sloping spur on the acromial undersurface. Patients with a keeled acromion are at significant risk of bursal-sided as well as full-thickness rotator cuff tears. All patients in this series had significant bursal-sided tears, and 12 of 20 patients (60%) had full-thickness rotator cuff tears associated with an acromial "keel." In addition, patients with a keeled acromion are younger (average age, 52.7 years) than would be expected for patients with rotator cuff disease. Often these patients are misdiagnosed, and effective treatment is delayed. Diagnosis of a keeled acromion relies on a high level of suspicion, as well as knowledge of clinical and radiographic signs consistent with an acromial keel. We discuss the clinical presentation, radiographic findings, associated rotator cuff pathology, as well as methods to avoid intraoperative technical errors when treating patients with an acromial keel. The goal is to raise awareness of this relatively uncommon but clinically important entity and to help guide the treating surgeon's therapeutic measures. PMID- 15346115 TI - Meniscal allograft transplantation--part I: background, results, graft selection and preservation, and surgical considerations. AB - Removal of the meniscus leads to progressive degenerative arthritis of the knee on a long-term basis. Therefore, meniscal allograft transplantation has been proposed as an alternative to meniscectomy. Although several experimental and clinical studies have documented that meniscal allografts show capsular ingrowth in meniscectomized knees, it remains to be established whether meniscal allograft transplantation can prevent degenerative changes after meniscectomy. Part 1 of this Current Concepts review will discuss the function, anatomy, and composition of the meniscus, followed by the history of surgery of meniscal tears and the healing of meniscal allografts in experimental and clinical studies. In addition, issues concerning preservation techniques, immunological reactions, sizing, disease transmission, indications, surgical technique, graft fixation, rehabilitation, and complications, will be taken into consideration. It can be concluded that the use of meniscal allografts in clinical practice has progressed to a point where relief of pain may be expected for the short-term. PMID- 15346117 TI - Arthroscopic removal of bullet fragments from the subacromial space of the shoulder. AB - This article describes a case of arthroscopic removal of a bullet fragment from the subacromial space. The bullet tore the rotator cuff and imbedded itself on the inferior surface of the acromion, producing pain and impingement. The bullet was removed, and the rotator cuff repaired arthroscopically. The patient noted complete relief of pain and improved range of motion. PMID- 15346118 TI - Arthroscopic anterior cruciate ligament distal graft rupture: a method of salvage. AB - We describe a rare case of anterior cruciate ligament (ACL) distal graft rupture in a high-demand rugby player. Fifteen months before this episode, he underwent an ACL reconstruction (autologous patellar tendon graft surgery) plus posterolateral reconstruction with direct suture and fascia lata augmentation. Radiographs revealed correct positioning of tunnels and fixation screws. Magnetic resonance imaging showed that the graft rupture was close to the tibial bone block and presented a signal compatible to the optimal graft incorporation. Surgery recording and clinical records were reviewed. No failures were found. After careful evaluation we concluded that the primary cause of failure was trauma. Based on these findings a salvage surgery technique was performed. Return to sport activities was allowed after four months when sufficient strength and range of motion had returned. Recent follow up (2 years 8 months postoperative) has shown an excellent result with a Lysholm score of 100, International Knee Documentation Committee (IKDC) score of 100, and a KT-1000 arthrometer reading of between 0 and 5 mm. The athlete has returned to his previous professional level. We believe this simple, specific, nonaggressive, and anatomic reconstructive technique may be used in the case of avulsion or distal detachment caused only by trauma and with a graft that is likely to heal. PMID- 15346119 TI - Patellar dislocation: arthroscopic patellar stabilization with anchor sutures. AB - There are many procedures to treat recurrent patellar subluxations and dislocations. Most of these procedures are open repairs for acute conditions or open reconstructions for chronic conditions, and both surgeries need large skin incisions. Although there are many arthroscopic procedures for meniscal and ligament injuries, there are very few arthroscopic procedures described for patellofemoral instability. In this report, we describe a new arthroscopic method to stabilize the patella for acute dislocation. Our technique includes an arthroscopic lateral release and a medial structure repair. We suggest that this procedure is minimally invasive and easier than traditional open procedures. Moreover, this may prevent patients from developing chronic instability of the patellofemoral joint. PMID- 15346120 TI - Identification of essential anatomic landmarks in performing arthroscopic single- and double-interval slides. AB - Contracted and immobile massive rotator cuff tears, which represent a small percentage of all massive tears, are believed by many to be irreparable. The development of arthroscopic single- and double-interval slides has allowed for the successful anatomic and tension-free repair of the majority of contracted and immobile massive rotator cuff tears. However, the safe and effective performance of interval slides requires clear and precise identification of certain anatomic landmarks, specifically the scapular spine and the base of the coracoid. This report describes how to arthroscopically identify and expose these anatomic landmarks. PMID- 15346121 TI - Margin convergence of the posterior rotator cuff to the biceps tendon. AB - Massive rotator cuff tears can often be repaired arthroscopically with satisfactory results. Tear pattern recognition, knot security, and loop security are essential in the repair of large and massive rotator cuff tears. In the case of a massive U-shaped tear, with a contracted immobile supraspinatus and a deficient rotator interval, a type of margin convergence technique can be used. This technique allows for the partial closure of the rotator cuff defect along with the proximal advancement of the posterior cuff by securing the posterior leaf of the cuff tendon to the tendon of the long head of the biceps brachii. Because of the mechanical effect of strain reduction in margin convergence, this procedure decreases the strain at the margin of the posterior cuff, thereby protecting its repair to bone. Our goal with this procedure is not to obtain healing of the cuff to the biceps, but to optimize the conditions for healing of the rotator cuff to bone. PMID- 15346123 TI - Long-term results after combined medial meniscal allograft transplantation and anterior cruciate ligament reconstruction: minimum 8.5-year follow-up study. PMID- 15346126 TI - A ganglion cyst that developed from the infrapatellar fat pad of the knee. AB - A ganglion cyst may be seen in all joints with varying frequency depending on location, but it is rare in the knee joint. A 30-year-old man presented with a mass in the anterolateral aspect of his left knee. Magnetic resonance imaging revealed an intra-articular tumor arising from the infrapatellar fat pad of the knee. This multilobular tumor showed low signal intensity on T1-weighted images and high signal intensity on T2-weighted images. Complete open resection was performed after arthroscopic examination. Histologic examination of the specimen confirmed the diagnosis of ganglion cyst. At the 1-year follow-up examination, the patient's left knee was completely asymptomatic with full range of motion and no recurrence. Ganglion cyst should be considered in the differential diagnosis of intra-articular masses. PMID- 15346127 TI - Donor-site morbidity after osteochondral autograft transfer procedures. AB - We report on 2 patients who had donor-site morbidity after an autogenous osteochondral grafting was performed. Both patients had fibrocartilage hypertrophy at the donor sites that contributed to knee pain and occasional locking; the second patient also had a lack of fibrocartilaginous regrowth with symptomatic residual osteocartilaginous defects. Additional arthroscopic surgery was required in both cases to trim the fibrocartilage. In addition, for the second case, a fresh osteoarticular allograft was used to transfer osteocartilaginous plugs back into the original knee donor sites due to continued knee pain. When performing an osteochondral autograft transfer, the benefits provided at the recipient site must be weighed against the possible donor-site morbidity that may result. PMID- 15346128 TI - Arthroscopic partial meniscectomy of a medial meniscus bucket-handle tear using the posteromedial portal. AB - Arthroscopic resection of irreparable bucket-handle tears of the medial meniscus is a commonly performed procedure. Adequate visualization of the posterior horn of the medial meniscus can be a challenging task with the conventional use of the anterior portal. An attempt to resect the posterior horn in a blind fashion may result in iatrogenic damage of the articular cartilage in the posterior compartment, over-resection of a remnant meniscus, or an insufficient resection of the torn fragment. We describe the use of the posteromedial portal for an accurate visualization and resection of the posterior attachment of a bucket handle tear for arthroscopic partial meniscectomy, as well as detection of other injuries that may be involved in the posteromedial compartment, while avoiding injury to other intra-articular structures during the arthroscopic procedure. We found that the use of the posteromedial portal is a safe and efficient method in removing a bucket-handle tear of the medial meniscus in one piece. PMID- 15346129 TI - Preliminary experience of fully arthroscopic repair of triangular fibrocartilage complex lesions. AB - Triangular fibrocartilage complex (TFCC) lesions are commonly repaired arthroscopically. Most of the interest in repairing TFCC lesions is focused on the development of an internal technique to avoid an extra incision of the skin. The most common techniques in use today are the inside-out or the outside-in, both of which have the disadvantage of requiring an additional opening to tie the suture. We suggest a personal method that allows an all-inside repair for type 1B lesions with a technique similar to the one used for rotator cuff repair in the shoulder. PMID- 15346131 TI - Neuropathology: the foundation for new treatments in spinal cord injury. AB - The first step essential in the search for a cure of human spinal cord injury (SCI) is to appreciate the complexity of the disorder. In this regard, it is not only the loss of ambulation but the sensory and autonomic changes that are equally important in recovery. In addition, there are the serious social emotional psychological and lifestyle effects of SCI which should also be taken into account. It is also true that no two SCI lesions are alike as each is the result of a SCI unique to that individual. Clinically of utmost importance is the segmental level of injury and whether it is complete, incomplete or discomplete (loss of all neurological functions below the injury but with physiological or anatomical continuity of Central nervous system tracts across the lesion). We are not concerned here with primary and secondary prevention or methods designed to limit the severity of the lesion after the event, important as they are, but with the requirements for a cure. Clearly, the greater the number of nerve fibers that can be preserved in the acute stage, the better will be the end result. Our focus at present is on the end-stage lesion with the aim of showing that a cure for SCI will depend upon establishing functionally useful central axonal regeneration and reestablishing physiological reconnections. Existing experimental methods are based on stimulating axonal regeneration by neutralizing inhibitory factors, adding positive trophisms and creating a permissive environment. Better results are obtained by bridging the gap with grafts of peripheral nerves or transplants of Schwann cells and genetically engineered fibroblasts. Recently, the potential for stem cells to enhance this process has created great interest. This is because of the ability of pluripotential cells to differentiate into neural tissue. A cure based on the physiopathology of SCI requires pyramidal, extrapyramidal, sensory, cerebellar and autonomic pathways to be regenerated with their appropriate neurotransmitters restored and reflexes integrated physiologically and in synchrony. In human SCI, there is a very long distance anatomically for axonal regrowth to occur in order to reach their relevant nuclei. This is because of continuing Wallerian degeneration. It also presumes that the target neurons are intact and that there has been no transneuronal degeneration above or below the lesion. Alternatively, in place of regenerated long axons, a multisynaptic pathway may be constructed from stem cells that have developed into neurons. Whether such a pathway would restore useful neurological functions is unknown. At present, the transplant and grafting research teams are exploring these possibilities in experimental animals. Moderate success in gaining axonal regeneration has been reported; however, it must be appreciated that the human lesion differs considerably from that of the experimental animal. In order to be successful, the neuropathology and neurophysiology of human SCI must be taken into account. The purpose of this review is to place the requirements for a cure, using stem cells, within the context of the neuropathology of human SCI. PMID- 15346132 TI - The Yin and Yang of the H-Channel and Its Role in Epilepsy. AB - Voltage-gated ion channels clearly are involved in the pathogenesis of epilepsy, with evidence implicating derangement of Na(+), K(+), and Ca(2+) voltage-gated channels, in both inherited and acquired forms of epilepsy ((1)). A newcomer to this list of ion channels involved in epilepsy is the hyperpolarization-activated cation channel or h-channel (otherwise known as I(h) or the pacemaker channel). This voltage-gated channel now is known to play a significant role in regulating neuronal excitability and recently has been shown to be modulated by seizures. Unlike other channels implicated in epilepsy whose function in normal neurons can clearly be labeled "excitatory" (Na(+) and Ca(2+)) or "inhibitory" (K(+)), the unique physiologic behavior of the h-channel allows it to both augment and decrease the excitability of neurons. Thus the role of I(h) in epilepsy, at present, is controversial and is a growing area of intense investigation ((2)(3)). PMID- 15346133 TI - T-Channel Defects in Patients with Childhood Absence Epilepsy. PMID- 15346134 TI - Susceptibility Gene for Juvenile Myoclonic Epilepsy Rings True. PMID- 15346135 TI - New Candidate Genes for Infantile Spasms and Mental Retardation. PMID- 15346136 TI - Modeling Poststroke Epileptogenesis by Middle Cerebral Artery Occlusion: Can It Work? PMID- 15346137 TI - Information Processing During Spike-Wave Discharges. PMID- 15346138 TI - GluR5 Kainate Receptors and Topiramate: A New Site of Action for Antiepileptic Drugs? PMID- 15346139 TI - Synchronized GABAergic Inhibition Drives Epileptiform Activity. PMID- 15346140 TI - How Does the Ketogenic Diet Work? PMID- 15346142 TI - Function in Cortical Malformations. PMID- 15346141 TI - Mortality Associated with Status Epilepticus. PMID- 15346143 TI - The Apolipoprotein E Epsilon (epsilon) 4 Allele Is Important for Trauma-related Epilepsy. PMID- 15346144 TI - Do New Antiepileptic Drugs Interact with Oral Contraceptives? PMID- 15346145 TI - Antiepileptic Drugs: Don't Sweat It! PMID- 15346146 TI - A New Anticonvulsant on the Horizon: How Does It Measure Up? PMID- 15346147 TI - Does Dual Pathology Always Require Dual Resection? PMID- 15346148 TI - Functional MRI Predicts Naming Deficits after Temporal Lobectomy. PMID- 15346150 TI - Treatment and Outcome of Psychogenic Nonepileptic Seizures. PMID- 15346149 TI - Transcranial Magnetic Stimulation in Epilepsy. PMID- 15346151 TI - Bilateral Frontoparietal Polymicrogyria Linked to Chromosome 16. PMID- 15346152 TI - Depression in Epilepsy Is Much More Than a Reactive Process. PMID- 15346153 TI - Drug Resistant Epilepsy: A Compliance Problem or an abnormality of Transport Proteins? PMID- 15346154 TI - Postmarketing Results: Useful, But Proceed with Caution. PMID- 15346155 TI - Intravenous Valproate for Pediatric Status Epilepticus. PMID- 15346156 TI - Can f MRI Substitute for the Wada Test? PMID- 15346157 TI - The Various Neuropathological "Faces" of Temporal Lobe Epilepsy. PMID- 15346158 TI - Progressive Damage in Epilepsy. PMID- 15346160 TI - Calcium and Autosomal Dominant Nocturnal Frontal Lobe Epilepsy (ADNFLE). PMID- 15346159 TI - SCN1A in SMEI, ICEGTC, and GEFS+: Alphabet Soup or Emerging Genotypic-Phenotypic Clarity? PMID- 15346161 TI - Dormant Basket Cell Hypothesis Revisited em leader Again. PMID- 15346162 TI - Does Hippocampal Neurogenesis after Kainate-induced Apoptosis In Neonatal Rats Replace Lost Neurons? PMID- 15346163 TI - Altered Sodium Channels Underlie Anticonvulsant Drug Insensitivity. PMID- 15346164 TI - Local Cooling: A Therapy for Intractable Neocortical Epilepsy. AB - Therapy for many of the neocortical epilepsies remains unsatisfactory. Recent research has demonstrated that focal cooling, using thermoelectric (Peltier) devices, may be capable of terminating, or possibly even preventing, some types of seizures. PMID- 15346166 TI - Epileptogenesis and Neurogenesis Share the Same Genes. PMID- 15346165 TI - Light at the End of the "TUNEL"? Role of Ceramide in Seizure-induced Programmed Cell Death. PMID- 15346167 TI - Role of Neuronal Loss in the Pathogenesis of Recurrent Spontaneous Seizures. PMID- 15346168 TI - Molecular Defect in Kindling Epilepsy. PMID- 15346170 TI - Psychogenic Nonepileptic Seizures Are Bad for Your Health. PMID- 15346171 TI - Cognitive Aging after ATL. PMID- 15346169 TI - Does Serotonin Play a Role in Epilepsy? AB - Studies in experimental models have suggested a potential role for serotonergic transmission in epilepsy, and interest in this research has been increased by the development of positron emission tomography (PET) ligands that can be used to study 5-hydroxytryptamine (5-HT) receptors and transporters. The serotonergic system is very complex. At least 13 distinct G protein-coupled 5-HT receptors and one ligand-gated ion channel receptor (5-HT(3)) are divided into seven distinct classes (5-HT(1) to 5-HT(7)) ((1)). The receptors vary widely in their distribution and effects, innervating vascular structures and gut smooth muscle as well as neuronal tissue. Several receptor subtypes may be relevant to epilepsy. PMID- 15346172 TI - Brain Abnormalities in Relatives of Patients with MCD. PMID- 15346173 TI - A novel method to analyze leukocyte rolling behavior in vivo. AB - Leukocyte endothelial cell interaction is a fundamentally important process in many disease states. Current methods to analyze such interactions include the parallel-plate flow chamber and intravital microscopy. Here, we present an improvement of the traditional intravital microscopy that allows leukocyte endothelial cell interaction to be studied from the time the leukocyte makes its initial contact with the endothelium until it adheres to or detaches from the endothelium. The leukocyte is tracked throughout the venular tree with the aid of a motorized stage and the rolling and adhesive behavior is measured off-line. Because this method can involve human error, methods to automate the tracking procedure have been developed. This novel tracking method allows for a more detailed examination of leukocyte-endothelial cell interactions. PMID- 15346175 TI - Review of ozone and temperature lidar validations performed within the framework of the Network for the Detection of Stratospheric Change. AB - The use of assimilation tools for satellite validation requires true estimates of the accuracy of the reference data. Since its inception, the Network for Detection of Stratospheric Change (NDSC) has provided systematic lidar measurements of ozone and temperature at several places around the world that are well adapted for satellite validations. Regular exercises have been organised to ensure the data quality at each individual site. These exercises can be separated into three categories: large scale intercomparisons using multiple instruments, including a mobile lidar; using satellite observations as a geographic transfer standards to compare measurements at different sites; and comparative investigations of the analysis software. NDSC is a research network, so each system has its own history, design, and analysis, and has participated differently in validation campaigns. There are still some technological differences that may explain different accuracies. However, the comparison campaigns performed over the last decade have always proved to be very helpful in improving the measurements. To date, more efforts have been devoted to characterising ozone measurements than to temperature observations. The synthesis of the published works shows that the network can potentially be considered as homogeneous within +/-2% between 20-35 km for ozone and +/-1 K between 35-60 km for temperature. Outside this altitude range, larger biases are reported and more efforts are required. In the lower stratosphere, Raman channels seem to improve comparisons but such capabilities were not systematically compared. At the top of the profiles, more investigations on analysis methodologies are still probably needed. SAGE II and GOMOS appear to be excellent tools for future ozone lidar validations but need to be better coordinated and take more advantage of assimilation tools. Also, temperature validations face major difficulties caused by atmospheric tides and therefore require intercomparisons with the mobile systems, at all sites. PMID- 15346176 TI - Measurements of atmospheric aerosol size distributions by co-located optical particle counters. AB - Ambient aerosol number concentrations and size distributions were measured in both indoor and outdoor environments using two identical co-located and concurrently operated optical particle counters (OPCs). Indoor measurements were performed in a research laboratory, whereas two different locations were used for outdoor measurements; the sampling duration exceeded 12 hours and one hour respectively. Results from the two OPCs have been presented for eight size classes between 0.5 and 20 [micro sign]m, represented by central value diameters 0.875, 1.5, 2.75, 4.25, 6.25, 8.75, 12.5 and 15 microm. Overall, for the six indoor and outdoor experiments conducted at different times of day, the mean particle count ratios from the two OPCs for the individual samples showed +/-20% variation for indoor experiments and +/-50% variations for outdoor experiments. Significant random departures of the mean ratios from unity at all size classes were noticed even for indoor sample periods exceeding 20 hours. However, the coefficient of determination (R(2)) for the plots of readings from the two OPCs indicated higher consistency for "fine" particles (0.5-3.5 microm) than for "coarse" particles (10-20 microm), with average R(2) > 0.8 and R(2) < 0.5 respectively. Poisson counting statistics help to explain the divergence in the latter case where number concentrations were very low for the outdoor experiments. However, it cannot explain the divergence for indoor measurements where the concentrations were much higher. Increasing the averaging period reduced the scatter, especially in size classes with low number concentration. However, this procedure may lead to over-smoothing of data for environments with rapidly changing number concentration. These results indicate that, when two such analysers are used for comparative studies, the divergence between their responses may generate significant values of source contribution or deposition flux, even for nominally similar aerosol populations. PMID- 15346177 TI - Indoor air quality during renovation actions: a case study. AB - A temporary renovation activity releases considerably high concentrations of particulate matter, viable and non-viable, into air. These pollutants are a potential contributor to unacceptable indoor air quality (IAQ). Particulate matter and its constituents lead, sulfate, nitrate, chloride, ammonium and fungi as well as fungal spores in air were evaluated in a building during renovation action. Suspended dust was recorded at a mean value of 6.1 mg m(-3) which exceeded the Egyptian limit values for indoor air (0.15 mg m(-3)) and occupational environments (5 mg m(-3)). The highest particle frequency (23%) of aerodynamic diameter (dae) was 1.7 microm. Particulate sulfate (SO(4)(2-)), nitrate (NO(3)(-)), chloride (Cl(-)), ammonium (NH(4)(+)) and lead components of suspended dust averaged 2960, 28, 1350, 100 and 13.3 microg m(-3), respectively. Viable fungi associated with suspended dust and that in air averaged 1.11 x 10(6) colony forming unit per gram (cfu g(-1)) and 92 colony forming unit per plate per hour (cfu p(-1) h(-1)), respectively. Cladosporium(33%), Aspergillus(25.6%), Alternaria(11.2%) and Penicillium(6.6%) were the most frequent fungal genera in air, whereas Aspergillus(56.8%), Penicillium(10.3%) and Eurotium(10.3%) were the most common fungal genera associated with suspended dust. The detection of Aureobasidium, Epicoccum, Exophiala, Paecilomyces, Scopulariopsis, Ulocladium and Trichoderma is an indication of moisture-damaged building materials. Alternaria, Aureobasidium, Cladosporium, Scopulariopsis and Nigrospora have dae > 5 microm whereas Aspergillus, Penicillium and Verticillium have dae < 5 microm which are suited to penetrate deeply into lungs. Particulate matter from the working area infiltrates the occupied zones if precautionary measures are inadequate. This may cause deterioration of IAQ, discomfort and acute health problems. Renovation should be carefully designed and managed, in order to minimize degradation of the indoor and outdoor air quality. PMID- 15346178 TI - Background concentrations of individual and total volatile organic compounds in residential indoor air of Schleswig-Holstein, Germany. AB - During a monitoring campaign concentrations of volatile organic compounds (VOCs) were measured in indoor air of 79 dwellings where occupants had not complained about health problems or unpleasant odour. Parameters monitored were the individual concentration of 68 VOCs and the total concentration of all VOCs inside the room. VOCs adsorbed by Tenax TA were then analysed by means of thermal desorption, gas chromatography and mass spectrometry. The analytical procedure and quantification was done according to the recommendation of the ECA-IAQ Working Group 13 which gave a definition of the total volatile organic compound (TVOC) concentration. Using this recommendation TVOC-concentrations ranged between 33 and 1600 microg m(-3) with a median of 289 microg m(-3). Compounds found in every sample and with the highest concentrations were 2-propanol, alpha pinene and toluene. Save for a few samples, all concentrations measured have been a factor 2 to 10 lower, compared to data from similar studies. Only a few terpenes and aldehydes were found exceeding published reference data or odour threshold concentrations. However, it has been found that sampling and analysing methods do have a considerable impact on the results, making direct comparisons of studies somewhat questionable. 47% of all samples revealed concentrations exceeding the threshold value of 300 microg TVOC m(-3) set by the German Federal Environmental Agency as a target for indoor air quality. Using the TVOC concentration as defined in the ECA-IAQ methodology is instrumental in assessing exposure to VOCs and identifying sources of VOCs. The background concentrations determined in this study can be used to discuss and interpret target values for individual and total volatile organic compounds in indoor air. PMID- 15346179 TI - Accumulation of polychlorinated naphthalenes in domestic animal related samples. AB - Concentrations of polychlorinated naphthalene (PCN) congeners were measured in domestic animal related samples such as feed ingredients, mixed feed and animal fat. Mean concentrations of total PCNs in feed ingredients ranged from 500 to 1500 pg g(-1) lipid wt with a high concentration found in fish meal. Total PCN concentrations were similar among mixed feeds, which ranged from 98 to 110 pg g( 1) lipid wt. The total PCN concentration in chickens was more than twice the amount in pigs. Tetra-CNs were the predominant homologues in all samples. Biomagnification of higher chlorinated PCN congeners, especially penta- and hexa CNs, was a few fold greater in chickens compared to pigs. The estimated concentrations of 2,3,7,8-tetrachloro dibenzo-p-dioxin equivalents (TEQs) of some selected PCNs in feed ingredients, mixed feeds, chickens and pigs were 0.008 to 0.063, 0.001 to 0.002, 0.033 and 0.011 pg g(-1) lipid wt, respectively. Based on predicted luciferase inducing potency for each PCN congener, the estimated PCN TEQs in feed ingredients and animal fat were similar to those that were estimated from selected PCNs. PMID- 15346180 TI - Analysis of HO-PCBs and PCP in blood plasma from individuals with high PCB exposure living on the Chukotka Peninsula in the Russian Arctic. AB - A trace analytical method is presented for the analysis of hydroxylated polychlorinated biphenyl metabolites (HO-PCBs) and pentachlorophenol (PCP) in human plasma. The described methodology is a modification of a previously validated method used for PCB and organochlorine pesticide analysis. The modified method enables the combined analysis of phenolic and neutral halogenated compounds. A tandem Florisil column is used for separating the HO-PCBs and PCP from the neutral fraction, instead of the more common chemical partitioning. In the same step the neutral fraction is purified for GC analysis. The extraction of the HO-PCBs and PCP was found to be highly dependent on sufficient acidification of the sample and the polarity of the extracting solvent. Analysis of plasma samples gave recovery rates for (13)C(6)-PCP and (13)C(12)-4-HO-CB 187 of 64 and 72%, respectively. The limit of detection ranged between 2-20 pg g(-1) plasma for the HO-PCBs and 5 pg g(-1) plasma for PCP. No matrix interferences were observed in the chromatograms. In plasma samples (n = 15) from the native Chukchi people in Uelen (Russian Arctic), a population with high PCB exposure, the median ratio of sum HO-PCBs to sum PCBs was as high as 0.4 and the sum HO-PCBs and PCBs were significantly correlated (r(2) > 0.7, p < 0.01). The median sum HO-PCBs (10 congeners) was 5920 pg g(-1) plasma with 4-HO-CB 107 as the dominating congener (median: 1670 pg g(-1) plasma). The median PCP level was measured at 642 pg g(-1) plasma. PMID- 15346181 TI - Origin of lead associated with different reactive phases in Scottish upland soils: an assessment made using sequential extraction and isotope analysis. AB - Soil samples (0-25 cm) have been taken annually since 1991 from three protected plots set up at an upland location at Glensaugh in Aberdeenshire, Scotland. The soils were analysed using the original BCR sequential extraction procedure and the lead isotopic composition was determined in each of the fractions, as well as the unfractionated soil using thermal ionisation mass spectrometry (TIMS). The lead concentrations in all the soils, including those well away from the road, were much higher than typical background values indicating that the whole area has been subject to deposition of anthropogenic lead. The distribution of lead between the different fractions was similar for the two non-roadside soils with most lead present in the oxidizable fraction. Although most lead in the roadside soil was also present in the oxidizable fraction, a substantial proportion (about 10%) was in the easily soluble fraction suggesting that roadside lead could be more mobile than lead in the other soils. Good reproducibility was obtained for the isotope analyses in all the fractions. The ratios calculated for the bulk soil from the ratios in the individual fractions agreed very closely with those measured directly in the unfractionated soil thereby demonstrating both reproducibilty and accuracy. The lowest (206)Pb/(207)Pb ratios were found in the roadside soil consistent with the recent deposition of petrol lead. The (206)Pb/(207)Pb ratios in all fractions of the other soils fell into a narrow band and it was necessary to use (204)Pb ratios to differentiate between lead in the extractable fractions and lead in the residual component. It is probable that lead in the non-roadside soils was deposited a considerable time ago and is characterised by a relatively high (206)Pb/(207)Pb ratio. Use of the (204)Pb ratios showed that the residual components in each of the three soils were isotopically distinct. PMID- 15346182 TI - Influence of ripening on pHstat leaching behaviour of heavy metals from dredged sediments. AB - The influence of oxidation and ripening on the kinetics of heavy metal release from anoxic sediments was investigated by means of porewater analysis and pH(stat) leaching tests. During simulated land disposal of an anoxic river sediment, a considerable heavy metal (Cd, Zn and Ni) release into the porewater was observed after 25 days of exposure of the sediment to the air. The release kinetics of Cd and Zn during pH(stat) leaching indicated that their release was related to the oxidation of Cd- and Zn-sulfides. It was not completely clear whether the release of Ni was due to the oxidation of Ni-sulfides or to the introduction of H(+)(generated by the oxidation of sulfides or introduced by titration). The release of Cu seemed to be explained by the oxidation of both Fe sulfides and Cu-sulfides. As such, pH(stat) experiments provided an indirect way to assess the mineralogy of the anoxic sediment. During ripening of the sediment, the release rate of Zn and Cd in pH(stat) leaching tests increased. For Ni, however, a similar release pattern was observed in the anoxic sediment and in the ripened sediments. Two weeks' drying at 100 degrees C resulted in an increase in the leachability of Cu. Nevertheless, at pH 4 and 6, the thermally treated sediment displayed a slower leaching of Cd and Zn than the ripened sediment. Although thermal treatment is often used to simulate ageing in soils, it cannot be used to simulate ripening of anoxic sediments. PMID- 15346183 TI - Photoremovable protecting groups based on electron transfer chemistry. AB - Photoremovable protecting groups (also known as photolabile protecting groups, phototriggers, or caged molecules) are functional groups that are attached to a molecule in such a way as to render the latter inactive. Exposure to light releases the protecting group, restoring functionality to the molecule. The use of photoremovable protecting groups (PRPGs) allows for precise spatial and temporal control of chemical reactions. Such groups have found use in many diverse applications, ranging from time resolved studies of physiological processes, to fabrication of spatially resolved combinatorial libraries of DNA. Recent research efforts have focused on designing protecting groups that are removed through photoinduced electron transfer (PET), rather than by direct photolysis. The PET strategy allows the light absorption step to be decoupled from the bond breaking step, thus permitting more control over the wavelengths of light used in the release process. The application of these types of protecting groups to the photochemical release of amines, alcohols, ketones, and carboxylic acids is described. PMID- 15346184 TI - Photodynamic therapy in oesophageal carcinoma: an overview. AB - Over the last five years, photodynamic therapy has become increasingly regarded as a viable option for palliative treatment of advanced oesophageal carcinoma, alongside more established modalities such as chemotherapy and radiotherapy. A large number of reports on the efficacy of photodynamic therapy as both a palliative and curative treatment for malignancy of the oesophagus have appeared in the 20 years since the first report on the use of this technique to treat oesophageal carcinoma, and this review aims to draw them all together to provide an overview of the current state of the field. PMID- 15346185 TI - Photochemistry of aryloxiranes: free radical 1,2-alkyl migrations in a constrained system. AB - The photolysis of alkylidenefluorene oxides resulted in free radical 1,2-alkyl migrations. PMID- 15346186 TI - Photooxidative degradation of beer bittering principles: product analysis with respect to lightstruck flavour formation. AB - Isohumulones, the main bittering agents in beer, are decomposed by light-induced reactions, thereby leading to radical precursors on the pathway to lightstruck flavour formation. Excited flavins, formed on visible-light irradiation, readily interact with isohumulones, as well as with reduced and oxidized derivatives thereof. From identification of both volatile and non-volatile reaction products thus formed, feasible degradation mechanisms are proposed. PMID- 15346187 TI - Photocatalytic reduction of aromatic azides to amines using CdS and CdSe nanoparticles. AB - We have shown that CdS and CdSe nanoparticles can act as very efficient and highly chemoselective photocatalysts for the reduction of aromatic azides to aromatic amines. In several cases, the reaction proceeds with quantum yields near 0.5, which approaches the theoretical maximum for a two-electron process. The wide scope of the reaction was confirmed with compounds containing electron withdrawing (-NO(2), CO(2)R, COR) and electron donating groups (-OMe, -R, -Cl) at the para-, meta-, and ortho-positions. Remarkably, the reaction is relatively insensitive to the electron demands of the substituent. However, azides with meta substituents give slightly lower yields than those with the same substituent at the ortho- or para-position. PMID- 15346188 TI - Evidence of homolytic and heterolytic pathways in the photodissociation of iminosulfonates and direct detection of the p-toluenesulfonyloxyl radical. AB - The mechanistic aspects of the photochemistry of several iminosulfonate photoacid generators (PAGs) have been studied based on product analysis, nanosecond laser flash photolysis, and determination of acid generation efficiencies. Our findings support a competition between homolytic and heterolytic N-O dissociation mechanisms. By measuring the efficiencies of acid generation for each PAG in the presence and absence of an ion quencher, we were able to roughly quantify the degree of branching between heterolytic and homolytic photocleavage pathways for each PAG. The p-toluenesulfonyloxyl radical was detected upon laser flash photolysis of several PAGs and was found to have a lambda(max) at 540 nm. By quenching the 540 nm transient with a variety of reactive species, the rate constants for reaction of the p-toluenesulfonyloxyl radical with these substrates were determined. The p-toluenesulfonyloxyl radical is shown to be a highly reactive species, which undergoes rapid hydrogen transfer and is a powerful oxidizer. PMID- 15346189 TI - Effect of thienyl groups on the photoisomerization and rotamerism of symmetric and asymmetric diaryl-ethenes and diaryl-butadienes. AB - Five symmetric (bis-substituted) and asymmetric (mono-substituted) analogues of E stilbene and EE-1,4-diphenylbutadiene, where one or both the side aryls are 2' thienyl or 3'-thienyl groups, have been studied by stationary and pulsed fluorimetric techniques, laser flash photolysis, conventional photochemical methods and theoretical calculations. The results obtained for these compounds and the comparison with those previously reported for three other compounds of the same series, allowed the effects of the position of the heteroatom and of the extension of the olefin chain on the excited state relaxation properties to be understood. The presence of one or two thienyl groups and their positional isomerism affect the spectral behaviour, the relaxation properties (radiative/reactive competition), the photoisomerization mechanism (singlet/triplet) and the ground state rotamerism. For the dienes containing the 3'-thienyl substituent(s), two rotamers were evidenced whose radiative and photochemical properties were obtained by selective excitation. PMID- 15346190 TI - Effect of oligothiophene substituents on the photophysical and photochromic properties of a naphthopyran. AB - A spectrokinetic study of the photophysical and photochemical properties of a series of photochromes bearing thienyl (T(1)), dithienyl (T(2)) and terthienyl (T(3)) groups linked to a naphthopyran in position 3 (sp(3) carbon) or 8 (naphtho moiety) has been carried out. The effect of the number and position of the thienyl groups on the excited state properties of the six compounds in the singlet and triplet manifold has been investigated by stationary and pulsed fluorimetric techniques, laser flash photolysis and kinetic analysis of the colouration/decolouration processes upon continuous irradiation. On increasing the length of the oligothiophene moiety, the contribution of fluorescence to the deactivation processes becomes detectable, while the photocolouration yield decreases. Moreover, only for compounds with T(3) groups was the triplet absorption observed by laser flash photolysis; its decay did not contribute to photocolouration. The competition between physical (fluorescence emission and intersystem crossing) and chemical relaxations markedly affects the behaviour of the compound bearing the T(3) group at the naphtho moiety, which shows the most efficient intersystem crossing and fluorescence emission while does not lead to photoproduction of the coloured open form. A parallel study of the NMR spectra of the compounds bearing one thiophene group allowed two and four photomerocyanine type photoproducts to be identified for the naphtho-substituted and pyran substituted compounds, respectively. The compound with a thiophene group at the naphtho moiety was studied in more detail at different temperatures. On the basis of the combined optical and NMR spectrometric results, a reasonable mechanism of its photochromic behaviour is proposed. PMID- 15346191 TI - Dynamics of the excited states of chromenes studied by fast and ultrafast spectroscopies. AB - In this paper, the photodynamics of three chromenes (2,2-spiro-adamantyl-7,8 benzo(2H)chromene, 2,2-diphenyl(2H)chromene and 2,2-diphenyl-5,6 benzo(2H)chromene) has been investigated by nano- and femtosecond time resolved techniques in hydrocarbon solutions at room temperature. Using pump-probe techniques, ultrafast steps of molecular dynamics characterizing the photoresponse of chromenes have been investigated: the breakage of the C-O bond was found to occur within a few hundred femtoseconds producing a short-lived transient that converts to the cisoid-cis open form in less than one picosecond and then to the metastable transoid-cis structure within a few picoseconds. The effect of different excitation wavelengths on the formation rate of the first shortest-lived transient of 5,6DPBC is in agreement with the model previously proposed to explain the wavelength dependence of the reaction and emission quantum yields in photostationary experiments. The results obtained in the nanosecond time domain show that the triplet marginally participates in the photoreaction for only one of the studied compounds (2,2-spiro-adamantyl-7,8 benzo(2H)chromene). PMID- 15346192 TI - Observation of the two triplet state conformations of alkyl phenylglyoxylates. AB - Step-scan time-resolved FT-IR spectra of alkyl phenylglyoxylates in hexane with a 4 cm(-1) spectral resolution reveal splitting of the transient absorption signal near 1650 cm(-1) into two closely located peaks with different lifetimes on the nanosecond time scale. This signal had been previously assigned to the triplet state of the starting material that gives rise to the alpha-hydroxyphenylketene. In the current article, evidence is presented to assign these two peaks to different triplet state conformers only one of which undergoes fast Norrish Type II photoelimination. This assignment was made on the basis of chemical reactivity, steric effects, and kinetic data. PMID- 15346193 TI - Forward (singlet-singlet) and backward (triplet-triplet) energy transfer in a dendrimer with peripheral naphthalene units and a benzophenone core. AB - The photochemical and photophysical behaviour of two dendrimers consisting of a benzophenone core and branches that contain four (4) and eight (5) naphthalene units at the periphery has been investigated in CH(2)Cl(2) solution (298 K) and in CH(2)Cl(2)/CHCl(3) 1:1 v/v rigid matrix (77 K). For comparison purposes, the photophysical properties of dimethoxybenzophenone (1), 2-methylnaphthalene (2) and of a dendron containing four naphthalene units (3) have also been studied. In both dendrimers 4 and 5, excitation of the peripheral naphthalene units is followed by fast (1.1 x 10(9) s(-1) at 298 K, > 2.5 x 10(9) s(-1) at 77 K for 5; 2.9 x 10(8) s(-1) at 298 K, 7 x 10(5) s(-1) at 77 K for 5) singlet-singlet energy transfer to the benzophenone core. On a longer time scale (>1 x 10(6) s(-1) at 298 K, >6 x 10(3) s(-1) at 77 K for 4; 3.1 x 10(7) s(-1) at 298 K, ca. 3 x 10(2) s(-1) at 77 K for 5) a back energy transfer process takes place from the triplet state of the benzophenone core to the triplet state of the peripheral naphthalene units. Selective excitation of the benzophenone unit is followed by intersystem crossing and triplet-triplet energy transfer to the peripheral naphthalene units. In hydrogen donating solvents, the benzophenone core is protected from degradation by the presence of the naphthalene units. In solutions containing Tb(CF(3)SO(3))(3), sensitization of the green Tb(3+) luminescence is observed on excitation of both the peripheral naphthalene units and the benzophenone core of 5. Upon excitation of the naphthalene absorption band (266 nm) with a laser source, intradendrimer triplet-triplet annihilation of naphthalene excited states leads to delayed naphthalene fluorescence (lambda(max)= 335 nm), that can also be obtained upon excitation at 355 nm (benzophenone absorption band). The results obtained show that preorganization of photoactive units in a dendritic structure can be exploited for a variety of useful functions, including photosensitized emission, protection from undesired photoreactions, and energy up-conversion. PMID- 15346194 TI - The role of propeptide in the refolding of human group IB phospholipase A2. AB - Human group IB phospholipase A2 (IB-PLA2) and its zymogen (proIB-PLA2) were purified from E.coli. Refolding was carried out by diluting the denatured forms of both IB-PLA2 and proIB-PLA2 with renaturation buffer in which the disulfide bonds were completely reduced. The refolding yield of proIB-PLA2 was increased by about 50% over that of the mature enzyme. The refolding of IB-PLA2 usually produced aggregates under normal conditions, as determined by light scattering. In addition, the unfolding experiments showed that the mature enzyme was more stable than the proenzyme toward denaturants in the presence of DTT. Results suggested that the N-terminal sequence rather than its conformation of human proIB-PLA2 played an important role in the refolding process. PMID- 15346195 TI - Recombinant bivalent vaccine against foot-and-mouth disease virus serotype O/A infection in guinea pig. AB - In this study, two DNA fragments encoding amino acid (141-160)-(21-140)-(141-160) of the VP1 of FMDV (foot-and-mouth disease virus) serotype O and (138-160)-(21 40)-(138-160) of the serotype A FMDV were chemically synthesized. These two tandem-repeat fragments were ligated and transfected into prokaryotic expression vector pTrcHis A to construct pTH-O-A. The other vector called pTH-O-scIgG-A was constructed similarly only that the two tandem-repeat DNA fragments were linked by the bovine-IgG heavy chain coding sequence. Guinea pigs immunized with the two bivalent vaccines pTH-O-A and pTH-O-scIgG-A showed both specific antibody activity and T cell proliferation responses. FMDV challenge tests showed that 85% and 70% of guinea pigs vaccinated twice with 200 mg of the fusion protein of pTH O-A were protected from FMDV serotype O and serotype A infection respectively. 70% and 57% of the guinea pigs immunized with the fusion protein of pTH-O-scIgG-A were protected from FMDV serotype O and serotype A infection respectively. PMID- 15346196 TI - Association between alcohol, smoking and HLA-DQA1*0201 genotype in psoriasis. AB - Psoriasis is a chronic skin disease triggered by genetic, environment or other risk factors such as infection, drugs, stress, moisture, alcohol, and smoking. A major psoriasis susceptibility locus at 6p21.3 has been identified. Further studies found that HLA-DQA1*0201 allele was associated with psoriasis. However, there were few data exploring an association between the environmental factors and susceptibility genes. In this study, the samples of 189 patients with psoriasis and 333 healthy controls were collected with their consent and were carried on analysis through polymerase chain reaction sequence-specific primer (PCR-SSP) method. The proportion of male psoriasis patients engaging in the smoking and alcohol was much higher than that of the control group (P<0.005). The HLA-DQA1*0201 allele was present at significantly higher frequency in the patients with psoriasis (OR=4.25, P<1.0 x 10(-6)). Association was found between smoking, alcohol and HLA-DQA1*0201 in male patients with psoriasis (OR>6.91, P<1.0 x 10(-4)). PMID- 15346197 TI - pH-dependent stability of EGX, a multi-functional cellulase from mollusca, Ampullaria crossean. AB - The cellulase activity and stability of EGX, a multi-functional cellulase previously purified from the mollusca Ampullaria crossean, was systematically studied under different pH. The pH induced con-formation and stability change of EGX have been investigated by using the intrinsic fluorescence, ANS fluorescence and CD spectrum. It has been found that the conformation and activity of this cellulase were strongly dependent on the pH. EGX was stable for both the enzyme activity and the conformation from pH 5.6 to pH 7.4. As shown by intrinsic and ANS fluorescence, no red shift of emission maximum occurred and a negligible intensity change was observed at pH 5.6-7.4. The activity of EGX remained about 80% in pH 5.6-7.4 and obviously decreased out of side the pH range. Urea-induced changes in EGX at pH 5.4 and pH 8.0 were measured by intrinsic fluorescence and CD spectrum. At pH 5.4, a significantly red shift of emission maximum occurred when the concentration of urea was 5 M compared to the concentration was 3 M at pH 8.0. The alpha-helix at pH 5.4 was 40.51% in the absence of urea and 31.04% in the presence of 4 M urea. At pH 8.0 the alpha-helix was 7.23% in the presence of 4 M urea. The data indicated that EGX was much susceptible to urea-induced unfolding at pH 8.0 and much stable at pH 5.4. The greater pH dependent stability of EGX may allow the enzyme to adequately catalyze the hydrolysis of cellulosic materials under natural or industrial extreme conditions. PMID- 15346198 TI - Cleavage of supercoiled circular double-stranded DNA induced by a eukaryotic cambialistic superoxide dismutase from Cinnamomum camphora. AB - A eukaryotic cambialistic superoxide dismutase (SOD) has been purified to homogeneity from mature seeds of the disease- and insect-resistant camphor tree (Cinnamomum camphora). Besides the known role of this SOD in protecting cells against oxidative stress, it can induce the cleavage of supercoiled double stranded DNA into nicked and linear DNA. It can not cleave linear DNA or RNA, demonstrating there is no DNase or RNase in the purified cambialistic SOD. Furthermore, the SOD can linearize circular pGEM-4Z DNA that is relaxed by topoisomerase I. This result indicates that the DNA-cleaving activity requires substrates being topologically constrained. The supercoiled DNA-cleaving activity of the cambialistic SOD can be inhibited by either SOD inhibitor (azide) or catalase and hydroxyl radical scavengers (ethanol and mannitol). The chelator of iron, diethylenetriaminepentaacetic acid (DTPA), also inhibits the supercoiled DNA-cleaving activity. These results show that the dismutation activity is crucial for the supercoiled DNA cleavage. The modification of tryptophan residue of the cambialistic SOD with N-bromosuccinimide (NBS) shows that these two activities are structurally correlative. The reaction mechanism is proposed that the hydroxyl radical formed in a transition-metal-catalyzing Fenton-type reaction contributes to the DNA-cleaving activity. In addition, the cleavage sites in supercoiled pGEM-4Z DNA are random. PMID- 15346199 TI - Trichostatin A extends the lifespan of Drosophila melanogaster by elevating hsp22 expression. AB - The level of acetylation of histones in nucleosomes is related to the longevity of yeast and animals. However, the mechanisms by which acetylation and deacetylation affect longevity remain unclear. In present study, we investigated the influence of histone acetylation modification on the expression of hsp22 gene and the lifespan in Drosophila melanogaster using histone deacetylase (HDAC) inhibitor Trichostatin A (TSA). The results showed that TSA could extend the lifespan of Drosophila melanogaster. Furthermore, TSA significantly promoted the hsp22 gene transcription, and affected the chromatin morphology at the locus of hsp22 gene along the polytene chromosome. Present data implicate that TSA may affect the lifespan of Drosophila through changing the level of histone acetylation and influencing the expression of hsp22 gene that is related to aging. PMID- 15346200 TI - Interaction between plasminogen activator inhibitor type-2 and pre-mRNA processing factor 8. AB - The plasminogen activator inhibitor type-2 (PAI-2) dependent apoptosis protection is due to the 33 amino acids fragment located between helix C and D of PAI-2, this fragment may interact with some unknown intracellular proteins. In this study we used the fragment between helix C and D of PAI-2 as a bait to perform a yeast two-hybrid screen using a cDNA library constructed with HeLa cells during apoptosis, and retrieved a clone encoding 94 amino acid residues of C-terminus of pre-mRNA processing factor 8 (PRPF8). Co-immunoprecipitation experiments confirmed that PAI-2 could interact with PRPF8 in vivo. PAI-2 could bind PRPF8 C terminal in both the inside and outside of nuclear. These results suggested that the interaction between these two proteins might not be involved in the apoptosis process. PMID- 15346201 TI - A new peptide with membrane-permeable function derived from human circadian proteins. AB - Basic peptides such as human immunodeficiency virus type 1 (HIV-1) Tat-(48-60) and Drosophila Antennapedia-(43-58) have been reported to have a membrane permeability and a carrier function for intracellular protein delivery. Based on the fluorescence microscopic observations of the vascular endothelial cells (ECV 304) and the primary cultured neuroglial cells, we found that human Clock protein DNA-binding peptide [residue 35-47, hClock-(35-47)] had a translocation activity very similar to Tat-(48-60). The cellular uptake of hClock-(35-47) increases with the increase of incubation time and concentration. The internalization effect at 4 degrees was same as that at 37 degrees C. Internalization of hClock-(35-47) was saturable and could be inhibited by the excess of the other MPPs. Moreover, the uptake of these peptides were significantly inhibited in the presence of heparan sulfate. These results strongly suggested that the hClock-(35-47) shared a common or very similar internalization pathway with other MPPs. Furthermore, we injected rat through the common carotid artery with hClock-(35-47)-FITC peptide, and cryostat sections of the brain were prepared and observed using a fluorescence microscope. Result showed that the peptide had the ability to translocate through the blood-brain barrier. It is promising to provide a new safe carrier for the intracellular and encephalic treatment. PMID- 15346202 TI - Preliminarily investigating the polymorphism of self-organized actin filament in vitro by atomic force microscope. AB - With the atomic force microscope (AFM), we preliminarily investigated the large scale structure of actin filaments formed in low concentration protein solution (5 microg/ml) via self-organization without the presence of any F-actin dynamic interfering factors (such as phalloidin) in vitro. It was found that the G-actin could be polymerized into ordered filamentous structures with different diameter from the slimmest filament of single F-actin to giant filament in tree-like branched aggregates. The observed polymerized actin filaments, to which our most intense attention was attracted, was discretely distributed and showed obvious polymorphism distinctly different from those in the presence of phalloidin or actin binding proteins (fimbrin, gelsolin, etc.) in previous experiments. Latter structures were mainly composed of single F-actin and/or multifilaments clearly consisting of several single F-actin. The experimental results clearly demonstrated that non-interference with the F-actin intrinsic dynamics in self organizing could lead to the polymorphism of actin filamentous structures, and further analysis implied that the disturbance of normal F-actin dynamics by many factors could prevent the emergence of structural polymorphism, more often than not, give rise to formation of specific structures instead and different interference would bring about various particular structures under certain conditions. PMID- 15346203 TI - Preparation of polyclonal antibodies of Rubisco large and small subunits and their application in the functional analysis of the genes. AB - Spinach Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) large (rbcL) and small (rbcS) subunits were separated by SDS-PAGE, and protein amount and purity were determined by Bradford assay. Polyclonal antibodies against rbcL and rbcS subunit were generated in female BALB/c mice and had no cross-reaction with each other. A total of 81 microg antigens were used and 0.3 ml anti-sera with titer of 1:5000 were yielded. The antibodies were also applicable to study rbcL and rbcS in tobacco plant Nicotiana benthamiana. Potato virus X vector pGR107 induced silencing of rbcS gene by Agrobacterium in Nicotiana benthamiana was performed. The expression level of rbcL and rbcS was lower in rbcS silenced plants than that in control plants as detected by the corresponding antibodies. This implied that the expression of rbcL was regulated by rbcS. PMID- 15346204 TI - Quality improvement guidelines for uterine artery embolization for symptomatic leiomyomata. AB - Uterine artery embolization (UAE) is assuming an important role in the treatment of women with symptomatic uterine leiomyomata worldwide. The following guidelines, which have been jointly published with the Society of Interventional Radiology in the Journal of Vascular and Interventional Radiology, are intended to ensure the safe practice of UAE by identifying the elements of appropriate patient selection, anticipated outcomes, and recognition of possible complications and their timely address. PMID- 15346205 TI - Management of tracheobronchial strictures. AB - Recently, the use of tracheobronchial stents has increased greatly sue to the advantages of easy placement and prompt airway relief. in addition, trachebronchial stents provide an alternative to open surgical procedures in select patients with benign tracheobronchial stenosis or obstruction, in particular those with tracheobronchial tuberculosis. This paper review the indications for tracheobronchial stent placement, appropriate stent selection, pre-procedure evaluation, and the results of stent placement for benign and malignant tracheobronchial strictures. PMID- 15346206 TI - Peritoneovenous shunt insertion for intractable ascites: a district general hospital experience. AB - Ascites often contributes to patient morbidity and discomfort. When refractory to medical management, it has been managed with repeated paracentesis, implantable ports for drainage, or diversion to the urinary bladder. Peritoneovenous shunt insertion has been a technique that was traditionally performed only by surgeons but is now within the realm of interventional radiologists. Its advantage is that protein-rich ascitic fluid is returned to the intravascular compartment. This retrospective study elaborates on the successes and problems encountered during shunt insertion in our first 13 patients. Two patients are well with a functioning shunt at 14 and 32 weeks. In 6 the shunt functioned until the patients' death from the underlying malignancy. Two required revision with variable success and in 2, shunt function could not be salvaged. There was one procedure-related mortality. Radiological insertion of these shunts is feasible and should be considered for selected patients. Options are available for assessment and salvaging of dysfunctional shunts. PMID- 15346207 TI - Percutaneous CT-guided core needle biopsy versus fine needle aspiration in diagnosing pneumonia and mimics of pneumonia. AB - The purpose of this study was to determine the diagnostic value of percutaneous core needle biopsy (CNB) relative to fine needle aspiration (FNA) in patients with pneumonia and pneumonia mimics. In this prospective study we present our experience with 48 thoracic FNAs and CNBs carried out on 48 patients with pneumonia and pneumonia mimics. Samples were obtained from all patients using both CNB (with an automated 18-G core biopsy needle and a gun) and FNA (with a 22 G needle). A specific diagnosis was made in 10/48 cases (20.83%) by FNA and in 42/48 (87.5%) by CNB. The main complications encountered were pneumothorax (n = 4) and hemoptysis (n = 2), yielding a total complication rate of 12.5%. We concluded that CNB using an automated biopsy gun results in a higher diagnostic accuracy for pneumonia and pneumonia mimic biopsies than FNA. Complications should be considered and proper patient observation should follow the procedure. PMID- 15346208 TI - Percutaneous balloon dilatation for the treatment of early and late ureteral strictures after renal transplantation: long-term follow-up. AB - We report our experience with percutaneous balloon dilatation (PBD) for the treatment of ureteral strictures in patients with renal allografts. Of the 422 consecutive patients after renal transplantation in our center 10 patients had ureteral strictures. An additional 11 patients were referred from other centers. The 21 patients included 15 men and 6 women aged 16 to 67 years. Strictures were confirmed by sonography and scintigraphy in all cases. Patients underwent 2 to 4 PBDs at 7-10-day intervals. Clinical success was defined as resolution of the stenosis and hydronephrosis on sequential ultrasound and normalization of creatinine levels. Patients were divided into two groups: those who underwent transplantation more than 3 months previously and those who underwent transplantation less than 3 months previously. PBD was successful in 13 of the 21 patients (62%). There was no statistically significant difference in success rate between the patients with early (n = 12) and those with late (n = 9) obstruction: 58.4% and 66%, respectively. No major complications were documented. PBD is a safe and simple tool for treating ureteral strictures and procedure-related morbidity is low. It can serve as an initial treatment in patients with early or late ureteral strictures after renal transplantation. PMID- 15346209 TI - Treatment of benign and malignant tracheobronchial obstruction with metal wire stents: experience with a balloon-expandable and a self-expandable stent type. AB - Over the last few years various types of metal wire stents have been increasingly employed in the treatment of both malignant and benign tracheobronchial obstruction. To date, however, few studies have investigated the in vivo properties of different stent types. We implanted 26 balloon-expandable tantalum Strecker stents (18 patients) and 18 self-expandable Wallstents (16 patients) into the tracheobronchial system of 30 patients with combined stenting in 4 patients. Mean age was 51 years (range: 0.5-79 years). Malignant disease was present in 23 patients, benign disease in seven patients. Both patients and individual stents were monitored clinically and radiographically. The probability of stents remaining within the tracheobronchial system, and of their remaining undislocated and uncompressed was calculated using Kaplan-Meier analysis for both stent types. Average stent follow-up time was 112 days until explantation and 115 days until patients' death or discharge. Kaplan-Meier analysis revealed a higher probability for the Wallstent to remain within the tracheobronchial system. Dislocation and compression occurred more rarely. Explantation, however, if desired, was more difficult compared to the Strecker stent. The Wallstent also led to the formation of granulation tissue, especially at the proximal stent end, frequently requiring reintervention. Both stent types proved to be effective therapeutic options in the management of obstructive tracheobronchial disease. The mechanical properties of the Strecker stent seem to be less favorable compared to the Wallstent but removal is easy. For benign disease, however, the Wallstent reveals limitations due to significant side effects. PMID- 15346210 TI - Treatment of benign tracheal stenosis utilizing self-expanding nitinol stents. AB - We assessed the results of self-expanding metallic stent insertion into benign proximal tracheal stenosis in patients not appropriate or unfit for surgical repair. Proximal benign tracheal stenoses had occurred in 11 patients (7 men, 4 women, mean age 68.8 years) after long-time intubation (n = 6), tracheostomy (n = 4), or chondropathia (n = 1). Fourteen self-expanding nitinol stents were placed in the patients under general anesthesia with endoscopical and fluoroscopical guidance. Stent insertion was successful in all cases and led to immediate relief of the morphological and functional airway obstruction. No immediate complications were noted. During the mean follow-up period of 67.5 weeks we observed one recurrent dyspnea 3 months after implantation and granuloma formation at the stent insertion site in another patient. Both complications were successfully treated with additional stent insertion in one case and laser resection of granulomas in the other. Self-expanding nitinol stents should be considered for the treatment of benign proximal tracheal obstruction in selected patients for whom surgical repair is contraindicated. PMID- 15346211 TI - Embolization of large gastric varices using vena cava filter and coils. AB - A 40-year-old male with alcoholic cirrhosis and portal hypertension presented with acute variceal hemorrhage. Abdominal CT scan and endoscopy revealed large gastric varices. The patient underwent a TIPS procedure. Portal venography demonstrated persistent filling of the large gastric varices with associated high flow spontaneous splenorenal shunt. Because of the large size of the varices, a Simon-Nitinol filter was used in conjunction with multiple embolization coils to enable successful occlusion of the varices. PMID- 15346212 TI - An unusual complication of a colonic wallstent. AB - We report a case of delayed perforation of normal colonic wall by the wire tips of an enteral Wallstent, which had successfully been used to treat a malignant obstruction of the sigmoid colon. Perforation occurred 5 days following insertion and despite surgery, resulted in fatality. Though perforation at the tumor site is a recognized complication during or following colonic stent placement, it is rare for the ends of the stent to perforate through nondiseased bowel wall. The site of the obstructing lesion and thus the position of the stent on a bend in the colon may be a contributory factor. PMID- 15346213 TI - Extracranial internal carotid artery aneurysms: report of a ruptured case and review of the literature. AB - Aneurysms of the extracranial carotid arteries (ECAA) are extremely rare. Schechter et al. [1] documented 835 cases in the literature up to 1977. One hundred and sixteen cases of ECAA have been documented in the Chinese literature since 1981, suggesting a higher prevalence of carotid aneurysmal disease in China than in the West. Four percent of all peripheral artery aneurysms are reported to be ECAA [2]. Those arising from the internal carotid artery (EICAA) are even more rare. Two recent reviews [3, 4] reported 24 and 25 cases of EICAA during 21 and 17 years, respectively, the majority of them is treated surgically. Our literature review revealed only a few true EICAA managed endovascularly, but none of them with a covered stent. We describe a rare such case of ruptured atherosclerotic EICAA which was treated percutaneously. PMID- 15346214 TI - Coil embolization of superior sagittal sinus dural arteriovenous fistula by transarterial intrasinus catheterization. PMID- 15346215 TI - Angiography in planning a posterior approach to the popliteal artery following reduction of an open posterior knee dislocation. PMID- 15346226 TI - Correlations between lactate and ventilatory thresholds and the maximal lactate steady state in elite cyclists. AB - We investigated the validity of different lactate and ventilatory threshold methods, to estimate heart rate and power output corresponding with the maximal lactate steady-state (MLSS) in elite cyclists. Elite cyclists (n = 21; 21 +/- 0.4 y; VO2peak, 5.4 +/- 0.2 l x min (-1)) performed either one (n = 10) or two (n = 11) maximal graded exercise tests, as well as two to three 30-min constant-load tests to determine MLSS, on their personal race bicycle which was mounted on an ergometer. Initial workload for the graded tests was 100 Watt and was increased by either 5 % of body mass (in Watt) with every 30 s (T30 s), or 60 % of body mass (in Watt) with every 6 min (T6min). MLSS was defined as the highest constant workload during which lactate increased no more than 1 mmol x l (-1) from min 10 to 30. In T30 s and T6 min the 4 mmol (TH-La4), the Conconi (TH-Con) and dmax (TH Dm) lactate threshold were determined. The dmax lactate threshold was defined as the point that yields the maximal distance from the lactate curve to the line formed by the lowest and highest lactate values of the curve. In T30 s also ventilatory (TH-Ve) and Vslope (TH-Vs) thresholds were calculated. Time to exhaustion was 36 +/- 1 min for T30 s versus 39 +/- 1 min for T6 min. None of the threshold measures in T30 s, except TH-Vs (r2 = 0.77 for heart rate) correlated with either MLSS heart rate or power output. During T6 min, power output at TH-Dm was closely correlated with MLSS power (r2=0.72). Low correlations were found between MLSS heart rate and heart rate measured at TH-Dm (r2=0.46) and TH-La4 (r2=0.25), respectively, during T6 min. It is concluded that it is not possible to precisely predict heart rate or power output corresponding with MLSS in elite cyclists, from a single graded exercise test causing exhaustion within 35-40 min. The validity of MLSS predicted from an incremental test must be verified by a 30 min constant-load test. PMID- 15346227 TI - Modification of the Wingate anaerobic power test for rowing: optimization of the resistance setting. AB - The purpose of the study was to determine the resistance factor that would elicit the highest peak 5 s and mean 30 s power output (PO) during a maximal 30 s anaerobic power test on a rowing ergometer. Thirty-one rowers (17 male and 14 female) were recruited based on the light-weight (LW) (6 male; age 23 +/- 6 yrs and 6 female; age 19 +/- 2 yrs) and heavy-weight (HW) (11 male; age 24 +/- 4 yrs and 8 female; age 27 +/- 8 yrs) rowing categories. Each group completed 5 randomized series of maximal 30 s sprints equivalent to the following forces: 58.9, 63.8, 68.7, 73.7 and 78.6 N for LW males; 83.5, 88.4, 93.4, 98.2 and 103.1 N for HW males; 29.4, 34.3, 39.2, 44.1 and 49.1 N for LW females; and 44.1, 49.1, 54.0, 58.6 and 63.8 N for HW females. The tests were performed on a Gjessing rowing ergometer modified to accommodate greater resistance settings and computer linked to obtain the necessary data. The peak 5 s and mean 30 s PO (W) were determined for each test. A relative load factor (RLF) for determining the amount of resistance to be applied was calculated based on body mass (BM). The RLF settings that elicited the highest peak 5 s PO were 0.109 and 0.102 kg. kg (-1) BM for LW and HW male rowers and 0.111 kg. kg (-1) BM and 0.076 kg. kg (-1) BM for LW and HW female rowers, respectively. The RLF settings for eliciting the highest mean 30 s PO were 0.102 and 0.095 kg. kg (-1) BM for LW and HW male rowers and 0.103 kg. kg (-1) BM and 0.068 kg. kg (-1) BM for LW and HW female rowers, respectively. A 30 second anaerobic test was also performed on a Concept II rowing machine for comparison and it was found to elicit a significantly lower peak 5 s but not 30 s PO in both male and female rowers. Our findings provide RLFs for assessing anaerobic power using a 30 s test in male and female rowers. As well, peak 5 s but not mean 30 s PO is underestimated using the Concept II rowing machine. PMID- 15346228 TI - Oxygen uptake-work rate relationship during two consecutive ramp exercise tests. AB - The performance of prior high intensity constant work rate (CWR) exercise significantly influences the gain of the fundamental oxygen uptake (VO2) response during subsequent high intensity CWR exercise. The purpose of the present study was to investigate whether equivalent effects could be elicited in the second of two bouts of exhaustive ramp exercise. We therefore hypothesised that a prior bout of exhaustive ramp exercise would increase the VO2-work rate (DeltaVO2/DeltaWR) slope during subsequent ramp exercise. Nine healthy males performed two ramp exercise tests to exhaustion on an electrically braked cycle ergometer separated by a 10-min period of cycling at 20 W. Pulmonary VO2 was measured breath-by-breath throughout both tests, and the mean response time (MRT) and the DeltaVO2/DeltaWR slope for exercise below the gas exchange threshold (GET) (S1), above the GET (S2), and over the S1 + S2 region (ST) were determined. Paired t-tests were used to analyse the data with significance accepted at p < 0.05. Blood [lactate] was higher at the onset of the second ramp test compared to the first (mean +/- SEM 1.2 +/- 0.1 vs. 6.2 +/- 0.7 mM; p < 0.01), but baseline VO2 was not significantly different between tests (0.93 +/- 0.05 vs. 0.99 +/- 0.06 L. min (-1)). The MRT (42 +/- 4 vs. 40 +/- 5 s) did not differ between tests, but the DeltaVO2/DeltaWR slope was steeper in the second ramp test for S2 (9.1 +/- 0.4 vs. 9.8 +/- 0.5 ml. min (-1). W (-1); p < 0.01) and ST (9.0 +/- 0.4 vs. 9.6 +/- 0.5 ml. min (-1). W (-1); p < 0.05). The demonstration that prior ramp exercise increases the DeltaVO2/DeltaWR slope during subsequent ramp exercise is consistent with the results of previous CWR studies and indicates that exercise economy is sensitive to the prior activity of the engaged muscles. PMID- 15346229 TI - Sleep deprivation, energy expenditure and cardiorespiratory function. AB - Participants in the sport of adventure racing often choose to go without sleep for a period of greater than 24 h while partaking in prolonged submaximal exercise. This study examined the effect of 30 h of sleep deprivation and intermittent physical exercise, on the cardiorespiratory markers of submaximal exercise in six subjects. Six subjects with the following physical characteristics participated in the study (mean +/- SD): age 22 +/- 0.3 years, height 180 +/- 5 cm, body mass: 77 +/- 5 kg, VO2peak 44 +/- 5 ml. kg (-1). min ( 1). Three subjects engaged in normal sedentary activities while three others cycled on a cycle ergometer at 50 % VO2peak for 20 min out of every two hours during thirty hours of sleep deprivation. One week later sleep deprivation was repeated with a cross over of subjects. Every four hours, subjects completed assessments of cardiorespiratory function during 50 % VO2peak cycling. A 3 x 8 repeated measures ANOVA revealed a significantly lower heart rate with sleep deprivation (p < 0.05), but no other significant effects (p > 0.05) on respiratory gas exchange variables. Neither sleep deprivation, nor a combination of sleep deprivation and five hours of moderate intensity cycling, appear to be limiting factors to the physiological capacity to perform submaximal exercise. PMID- 15346230 TI - The -55 C/T polymorphism within the UCP3 gene and performance during the South African Ironman Triathlon. AB - Uncoupling protein 3 is believed to be involved in total body energy expenditure, including the regulation of fat and glucose metabolism. These biochemical processes may distinguish top ultra-endurance triathletes from slower competitors. The aim of this study was to determine whether the uncoupling protein 3 gene is associated with the performance capacity of ultra-endurance Ironman triathletes. Two triathlete groups consisting of the 89 fastest and 89 slowest Caucasian, male triathletes who completed either the 2000 or 2001 South African Ironman triathlon events were genotyped for the -55 C/T polymorphism within the uncoupling protein 3 gene. A control group consisting of 92 Caucasian males who had not trained for or participated in an ultra-endurance athletic event was also genotyped. There was no significant difference in the genotype (CC, CT and TT) frequency distribution of the -55 C/T polymorphism within the uncoupling protein 3 gene between the fast triathlete, slow triathlete and control groups. In addition, no significant differences were observed between the frequencies of the C and T alleles between the three groups. The two triathlete groups were combined and grouped according to their genotype. No particular genotype or allele was associated with the time taken by the triathletes to complete the entire triathlon, or either the swim, cycle or run legs of the event. Thus no association was found between the -55 C/T polymorphism within the uncoupling protein 3 gene and the ultra-endurance performance of triathletes who completed either the 2000 or 2001 South African Ironman triathlons. PMID- 15346231 TI - Effects of downhill treadmill running on uncoupling protein 3 mRNA expression. AB - Eccentric biased exercise has been reported to elicit more muscle injury than concentric or isometric exercise and potentially generate increased oxidative stress one to two days post exercise. Increased oxidative stress has been shown to up-regulate the expression of UCP3 mRNA. The aim of this study was to investigate the effects of downhill running on skeletal muscle UCP3 mRNA expression. Twenty-four male Sprague Dawley rats were randomly assigned to run continuously for 30 minutes (30-C, n = 6), or run six 5-minute bouts separated by rest periods of 2 minutes (2-R, n = 6), 4 minutes (4-R, n = 6), and 6 minutes (6 R, n = 6) on a 16 degree declined treadmill at a speed of 16 m. min (-1). Sham control animals (n = 8) were placed in a treadmill chamber during the 30-minute run session. Semi-quantitative RT-PCR was conducted to evaluate UCP3 mRNA levels in the plantaris, a muscle used eccentrically during downhill running and tibialis anterior, a muscle which undergoes very little eccentric muscle contraction during this exercise. The level of gene expression was normalized to 18 S ribosomal mRNA expression from the same PCR product. Results are reported as mean +/- standard error. UCP3 of the plantaris muscles from 2-R animals (2.36 +/- 0.13) was significantly greater than UCP3 of the plantaris from control animals (1.72 +/- 0.13), p < 0.05. UCP3 of the tibialis anterior from the continuous group (1.51 +/- 0.17) was significantly less than the UCP3 of the tibialis anterior of the control group (2.09 +/- 1.4), p < 0.05. These data suggest that downhill treadmill running is associated with an increase in UCP3 mRNA expression in the plantaris muscle. These results indicate that exercise which is biased toward eccentric exercise may up-regulate UCP3 mRNA during the period post exercise when muscle damage and repair is elevated. PMID- 15346232 TI - Composition and turnover of contractile proteins in volume-overtrained skeletal muscle. AB - The purpose of this study was to find the composition shift of myosin heavy chain (MyHC) isoforms in overtraining in fast- and slow-twitch skeletal muscles and different changes in MyHC isofom composition, synthesis and turnover rate between 4-week and 6-week overtraining. Male Wistar rats were randomly assigned to 4-week and 6-week endurance training, 4-week and 6-week overtraining groups. Plantaris (Pla), extensor digitorum longus (EDL), and soleus (Sol) muscles were studied. Daily excretion of 3-methylhistidine (3-MeHis) pool as an indicator for protein degradation increased in the 4-week and 6-week overtraining group to 4.04 +/- 0.21 and 4.32 +/- 0.23 %/day subsequently in comparison with the control group (2.16 +/- 14 %/day, p < 0.001). In Pla muscle MyHC I isoform synthesis rate was 33 200 +/- 2150 (after 6-week overtraining 27 100 +/- 1800, p < 0.05), IIa 32 600 +/- 2100; IId 27 300 +/- 1890 and IIb isoform 20 100 +/- 1600 (after 6-week overtraining 15 500 +/- 1400, p < 0.05) dpm/M leucine/min. Actin synthesis rate increased in fast-twitch muscles during 4- and 6-week overtraining, and in soleus muscle during 6-week overtraining. In EDL and Sol muscle MyHC isoform composition during 6-week overtraining did not change significantly. During the 6-week overtraining the relative content of MyHC I and IIb isoforms decreased and IIa and IId isoforms increased in Pla muscle. The initial increase of MyHC IIb isoform after 4-week overtraining shows the higher stability of this isoform in comparison with MyHC I isoform in fast-twitch muscles during high volume exercise. PMID- 15346233 TI - D-amphetamine-induced hydrogen peroxide production in skeletal muscle is modulated by monoamine oxidase inhibition. AB - The aim of this paper was to study the influence of d-amphetamine administration as a sympathomimetic drug on the synthesis of hydrogen peroxide (H2O2) in mouse soleus muscle and to investigate the modulating effects of pargyline, an inhibitor of monoamine oxidase (MAO) in this context. Charles River mice were assigned to four groups: Control, d-amphetamine treated, pargyline treated, and amphetamine + pargyline treated. Their soleus muscles were removed 0, 15, 30, 60, and 120 min after treatment. The amount of hydrogen peroxide formation within the muscles was estimated using an indirect method. The control data showed a continuous production of hydrogen peroxidase. Pargyline administration lead to an initial increase of H2O2 production that later faded below control levels. Administration of amphetamine finally stimulated H2O2 production much above control levels. When combining amphetamine and pargyline treatment, H2O2 production was accelerated in the initial phase but dropped to control levels at 30 min. It is concluded that in skeletal muscle MAO is an important source of hydrogen peroxide production triggered by amphetamine administration and that this tissue plays a hitherto not described role in oxidizing circulating biogenic monoamines. PMID- 15346234 TI - Fatigue and recovery after high-intensity exercise part I: neuromuscular fatigue. AB - The contribution of central and peripheral factors to muscle fatigue were quantified following a high-intensity uphill running exercise. Eight male volunteers performed an intermittent exercise at 120 % of maximal aerobic speed on a treadmill with an 18 % grade. Electrically evoked and voluntary contractions of the knee extensors and EMG of the two vastii were analyzed before and immediately after the high-intensity exercise. Isometric maximal voluntary contraction decreased slightly (-7+/-8 %; p < 0.05) after exercise but no changes were found in the level of maximal activation or in the torque produced by a 80 Hz maximal stimulation applied to the femoral nerve. Following exercise, the single twitch was characterized by lower peak torque, maximal rate of force development, and relaxation (-28+/-11%, -25+/-12%, -31+/-15% respectively, p < 0.001), and higher surface of the M-wave for both vastii. The ratio between the torques evoked by 20 Hz and 80 Hz stimulation declined significantly (-22+/-10%, p < 0.01) after exercise. These findings indicate that muscle fatigue after high intensity running exercise is due to significant alteration in excitation contraction coupling and that this type of exercise does not induce significant central fatigue or changes at the crossbridge level. PMID- 15346235 TI - Prolonged asystolia in a young athlete: a case of sinus arrest during REM sleep. AB - A young asymptomatic male athlete came to our laboratory to be enrolled in a research protocol on physical fatigue. Routine clinical and cardiological evaluations including echocardiogram were in the normal range. Several consecutive cardiopulmonary tests showed a fairly good tolerance to exercise, with no symptoms even when the effort was abruptly arrested. On the other hand, Holter ECG recordings showed long nocturnal sinus pauses. As he was absolutely asymptomatic and free from any structural heart disease, he underwent a follow-up with repeated Holter monitorings for one year. During this period he decided on his own to stop practising sports; in spite of this sharp reduction in his overall physical activity, consecutive Holter monitorings showed that the sinus pauses were progressively increasing in duration (up to 9.2 seconds). With the hypothesis of a malignant vagotonia, he underwent a tilt test; however, we could not elicit any pauses or symptoms. The pauses grew longer over time; a endocavitary electrophysiologic test was performed, which showed no evidence of disease. To rule out the hypothesis of a sleep apnoea syndrome, he also underwent a polysomnography, including EEG, eye movement electromyography, arterial blood oxygen saturation and thoracic impedance: no alterations were detected with the exception of the sinus pauses, which appeared to be strictly linked to REM sleep, as suggested by the concurrent increase in rapid eye movements and desynchronized EEG. We hence made a diagnosis of sinus arrest during REM sleep (SAdRS), a very uncommon disease belonging to the parasomnias. Pauses were then quantified for one month by implanting a ECG loop recorder. As the patient became more and more upset and worried, and the pauses increased to nearly 12 seconds, we decided to implant a pacemaker, which is the only therapeutic option established in the literature for patients with SAdRS. PMID- 15346236 TI - Cortisol--essential adaptation hormone in exercise. PMID- 15346237 TI - A comparison of rowing technique at different stroke rates: a description of sequencing, force production and kinematics. AB - Low back pain is the commonest musculoskeletal complaint in rowers. Research into the relationship between rowing technique, the forces generated during the rowing stroke and the kinematics of spinal motion are increasing, but to date none have investigated the impact of different rowing intensities on this relationship. A technique has been developed using an electromagnetic motion system and strain gauge instrumented load cell to measure spinal and pelvic motion and force generated at the handle during rowing on an exercise rowing ergometer. Using this technique ten collegiate male rowers (mean age 22.1+/-2.8 years) from local rowing clubs were investigated. The test protocol consisted of rowing on an ergometer at three different stroke ratings; 17-20 strokes per minute; 24-28 strokes per minute; and 28-36 strokes per minute. Each rating was held for four minutes, with a five-minute rest between each rating. Marked changes in the force output curve and lumbopelvic kinematics were observed at the different rowing intensities. Although there was no change in the magnitude of peak torque generated during the different rating, there was a marked shift in when this occurred during the stroke. In terms of kinematic changes, these centred around changes in pelvic rotation at the catch and finish stages of the stroke with significantly less anterior rotation occurring at the catch position at higher rowing intensities. To conclude, this study suggests that rowing kinematics and force profiles do change at higher rowing intensities. These changes may be an important factor with respect to injury mechanisms, however, further work is required at an elite level. PMID- 15346238 TI - Immune system alteration in response to increased physical training during a five day soccer training camp. AB - Leukocyte and monocyte subpopulations were investigated in ten elite male soccer players before and after a 5-day training camp. It was hypothesized that with increased training intensity and duration, the immune system would show signs of depression. Blood samples were taken at rest before and after the training camp and cell surface antigens were investigated by four-colour flow cytometry. After five days of intensified training, there was a significant decrease in the number of T helper, T cytotoxic and B cells, the expression of CD11 b on leukocytes increased and the NK cell population did not change significantly. It is concluded that after a period of intensified training, soccer players may experience decreased T and B cell numbers in circulation, possibly affecting their capability to activate the immune system and resist infections. However, in contrast to the acute decrease in the number of circulating NK cells commonly observed after physical exercise, no change in this cell population was observed at rest after a period of intensified physical training. Exercise-induced immunological changes were highly differentiated between different leukocyte subpopulations. PMID- 15346239 TI - Chronic hepatitis C. PMID- 15346240 TI - The natural history of hepatitis C. AB - Hepatitis C virus (HCV) is the most common cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma (HCC) in the United States and infects an estimated 170 million people worldwide. In the United States, overall hepatitis C prevalence is 1.8% and is higher in African Americans and Hispanics than it is in white Americans. The age distribution places 65% of all subjects with anti-HCV antibodies between 30 and 49 years of age. Before serological screening of blood products was initiated, hepatitis C was most commonly transmitted by transfusion of contaminated blood products, but now it is acquired primarily via intravenous drug use. Only 10 to 25% of infected adult patients spontaneously resolve their infection, and the remaining 75% remain persistently viremic and often asymptomatic. Progression of liver disease in 20 to 30% of patients can lead to compensated and eventually decompensated cirrhosis or hepatocellular carcinoma, or both. This article looks briefly at the epidemiology and natural history of hepatitis C. PMID- 15346241 TI - Use of diagnostic testing for managing hepatitis C virus infection. AB - Serological and virological testing for the hepatitis C virus (HCV) is essential in the management of patients with chronic HCV infection. Recent advances in molecular virology tests and the development of a new international unit standard have greatly simplified the use and interpretation of these tests. In the absence of treatment, baseline serum HCV RNA does not change over time and does not correlate with the severity of hepatic inflammation or fibrosis on liver biopsy. In contrast, changes in serum HCV RNA level are important predictors of virological response to treatment with pegylated interferon and ribavirin. PMID- 15346242 TI - Treatment of hepatitis C virus: the first decade. AB - Interferon alfa therapy emerged as an early treatment option for patients with chronic hepatitis C. This therapy, however, fails to produce a sustained virological response in most patients. Various host and viral baseline characteristics, some of which include hepatitis C virus genotype, viral load, presence of cirrhosis, and patient age, affect the response to interferon therapy. The addition of ribavirin to interferon therapy significantly improves long-term virological response in treatment-naive patients and is also more effective than repeat interferon therapy is in patients who fail to initially achieve sustained virological or biochemical responses. However, ribavirin can induce reversible hemolytic anemia, and combination therapy with a ribavirin/interferon regimen is not tolerated as well as interferon is alone. Pegylated interferons used alone or in combination with ribavirin provide improved treatment options for different patient groups with chronic hepatitis C. PMID- 15346243 TI - Predictors of response to therapy for chronic hepatitis C. AB - Pretreatment prediction of successful interferon-based therapy is less accurate than is prediction based on serum hepatitis C virus (HCV) RNA measurements taken during the first few weeks of therapy. Virological response, defined as loss of detectable virus RNA during the first weeks of treatment, identifies interferon sensitive virus strains. Studies on viral decline kinetics have shown that interferon-based therapies produce a two-phase reduction in viral RNA levels. Sustained virological response is associated with rapid biphasic viral decline kinetics, resulting in elimination of virus from the blood within 4 to 12 weeks. Rapid viral elimination correlates positively with low incidence of relapse in end-of-treatment virological responders and is, therefore, an important parameter for determining duration or type of therapy. This article reviews the various patient and viral factors that help predict response to various interferon-based therapies, ranging from interferon alone to the new pegylated interferons. PMID- 15346244 TI - Development and pharmacokinetics and pharmacodynamics of pegylated interferon alfa-2a (40 kD). AB - Polyethylene glycol (PEG) moieties can be attached to proteins to improve their pharmacological properties. Variables such as the molecular weight and conformation of PEG chains have a bearing on the conjugate's properties and should be considered when developing pegylated proteins with desired characteristics. Recombinant interferon (IFN) alfa has been used, with limited success, to treat patients infected with hepatitis C virus. The recommended thrice-weekly administration of IFN alfa does not maintain sustained plasma concentrations of the drug, thereby adversely affecting the potential virological response. Pegylation alters the pharmacokinetic properties of IFN alfa and allows for once-weekly administration. Pegylated IFNs contain either linear PEG chains of small molecular weight, as is pegylated IFN alfa-2b (12 kD), or larger branched moieties, as in pegylated IFN alfa-2a (40 kD). There are pharmacokinetic and pharmacodynamic differences between the two IFNs. The much-increased sustained virological response rates observed with pegylated IFN alfa-2a (40 kD) and pegylated IFN alfa-2b (12 kD) support the rationale for pegylation of IFN. PMID- 15346245 TI - Pegylated interferon monotherapy for chronic hepatitis C. AB - Interferon (IFN) alpha-2a has been attached to a branched 40-kD PEG molecule and IFN alpha-2b to a linear 12-kD PEG molecule leading to elimination half-lives of approximately 75 and approximately 30 hours, respectively. In one pivotal trial, 531 patients with chronic hepatitis C were assigned to receive either 180 microg of pegylated IFN alpha-2a once weekly for 48 weeks or 3 x 6 mIU standard IFN for 12 weeks, followed by 3 x 3 mIU for 36 weeks. Sustained virological response rates were 39 and 19% for pegylated and standard IFN alpha-2a, respectively. In a second trial in patients with hepatitis C virus (HCV)-associated cirrhosis and bridging fibrosis, sustained virological response rates were 8% (3 x 3 mIU IFN three times a week), 15% (90 microg PEG-IFN alpha-2a four times a week), and 30% (180 microg PEG-IFNalpha-2a four times a week). In a third trial, 1219 patients with chronic hepatitis C were randomly assigned to receive either standard IFN alpha-2b (3 x 3 mIU) or once weekly pegylated IFN alpha-2b (0.5, 1.0, or 1.5 microg/kg). Sustained virological response rates were highest in the 1.0 microg/kg dose and achieved 25% compared with 12% in the standard IFN group. In conclusion, each regimen of pegylated IFN given once weekly is more effective than a regimen of standard IFN given three times weekly. PMID- 15346246 TI - Treatment of chronic hepatitis C infection with peginterferons plus ribavirin. AB - Interferon monotherapy provided the first hope for patients with chronic hepatitis C that the virus could be permanently eradicated. An important development in treating this disease was the recognition that the effects of interferon could be greatly enhanced by combining it with ribavirin, a nucleoside analogue. This combination regimen essentially doubled the sustained virological response rates seen with interferon alone. Recently, modified forms of interferon have been developed that-when used in combination with ribavirin-demonstrate even better efficacy. Thus, peginterferon alfa-2a (Hoffman La-Roche, Nutley, NJ) and peginterferon alfa-2b (Schering-Plough, Kenilworth, NJ) are the latest innovations for the treatment of chronic hepatitis C. The addition of a polyethylene glycol molecule to the native interferon protein favorably alters the pharmacokinetic profile, allowing for once-weekly administration, and leads to superior efficacy compared to standard interferon preparations. This article reviews the data from clinical trials of peginterferon alfa-2a and peginterferon alfa-2b. PMID- 15346247 TI - Histological response to interferon alfa-based therapies in hepatitis C. AB - Histological response is defined as an improvement in the severity of liver inflammation with or without an improvement in fibrosis in patients following treatment for an acute or chronic liver disorder. Histological response occurs in patients with chronic hepatitis C virus (HCV) infection who have had a sustained virological response following treatment with interferon (IFN) therapy and in a percentage of nonresponders. Continuing IFN long term as maintenance therapy has been shown to prevent histological progression in patients who had both a marked decline in serum HCV RNA level during treatment and a histological response on repeat histological analysis. Prospective controlled trials are currently evaluating the benefits of low-dose maintenance therapy with pegylated IFN in HCV patients who have failed to achieve sustained virological response. In the future, the development of serum markers of fibrosis may allow for monitoring of hepatic fibrosis and histological response to therapy without the need to perform repeat liver biopsy. PMID- 15346248 TI - Hepatitis C virus in the setting of HIV or hepatitis B virus coinfection. AB - Because of shared routes of transmission, coinfection with hepatitis C virus (HCV) or hepatitis B virus (HBV), or both, is common among HIV-infected persons, affecting approximately 15 to 30% and 10 to 15% of patients, respectively. Advances in antiretroviral therapy have improved the life expectancy of patients infected with HIV, and, as a consequence, HCV-related liver disease has emerged as a significant comorbid disease among such patients. Concurrent HIV infection may be associated with higher serum HCV RNA levels, accelerated progression of hepatic fibrosis, increased risk of end-stage liver disease, hepatocellular carcinoma and death among persons coinfected with hepatitis C. Similarly, coinfection with HCV and HBV may lead to more severe liver disease and greater risk of hepatocellular carcinoma (HCC) than does HCV infection alone. Although definitive randomized controlled trials are not yet completed, current guidelines recommend the use of pegylated interferon alfa plus ribavirin for the treatment of chronic HCV in eligible HIV-infected persons. Conversely, the optimal treatment of chronic HCV in persons with chronic HBV infection has not been defined but may include pegylated interferon alfa plus ribavirin, with or without additional antiviral agents, such as lamivudine or adefovir, or both. PMID- 15346249 TI - Hepatitis C infection associated with renal disease and chronic renal failure. AB - Screening of blood products and attention to stricter infection control measures in hemodialysis units have reduced the incidence of hepatitis C virus (HCV) infection among dialysis patients. HCV can be transmitted via transplanted organs. Renal transplantation may accelerate the course of liver disease, which has an impact on patient and graft survival. Interferon (IFN) alfa monotherapy has produced promising results during treatment but disappointing long-term results in patients with HCV-associated glomerulonephritis. Dialysis patients with HCV infection respond well to IFN-based therapy, and there appears to be clinical benefit in clearing HCV in renal transplantation candidates. Larger prospective trials are required to fully determine the role of IFN in these patient groups, including the potential use of IFN plus ribavirin and pegylated IFNs. IFN therapy in renal transplantation patients is not recommended because of potential graft rejection. PMID- 15346250 TI - Natural history and management of hepatitis C infection after liver transplantation. AB - With almost universal virological recurrence and a projected several-fold increase in HCV-related end-stage liver disease within the next decade, a clear understanding of the natural history of posttransplant HCV infection and optimization of the management of recurrent HCV infection are imperative. This article describes the natural history, risk factors, and current strategies for the amelioration of posttransplant HCV infection. PMID- 15346251 TI - Role of amantadine and other adjuvant therapies in the treatment of hepatitis C. AB - There is room for improvement in the treatment of chronic hepatitis C with standard interferon (IFN) alfa. In the search for treatment adjuvants, the antiviral compound ribavirin has been found to significantly increase sustained virological response. Despite this improvement, the rate of "cure" remains low at approximately 40%, thus stimulating the search for additional adjuvants. In 1997, it was suggested that amantadine monotherapy could be used successfully to treat patients with chronic hepatitis C who had previously failed IFN alfa therapy, but ensuing studies could not support these findings. Instead, researchers have studied amantadine as an adjuvant to either IFN alfa alone or IFN alfa plus ribavirin, and promising results have been published. In this article, the author reviews the role of amantadine alone or as part of combination therapy regimens for chronic hepatitis C and briefly looks at the use of other agents as potential adjuvants. PMID- 15346252 TI - Past, present, and future hepatitis C treatments. AB - Conventional interferon (IFN) alfa has been used for many years in the treatment of chronic hepatitis C. However, few patients achieve sustained virological responses with IFN. Combining IFN with ribavirin improves efficacy considerably, but at the expense of diminished tolerability attributable to ribavirin. Pegylated interferons have improved pharmacokinetic profiles, may be administered once weekly, and are more effective than IFN is alone or in combination with ribavirin. In addition to enhanced efficacy, pegylated interferon alfa-2a (40 kD) also improves health-related quality of life during therapy compared with IFN based therapy. New adjuvant agents have the potential to further improve sustained response rates and tolerability; however, pegylated interferons will likely remain the backbone of therapy in the foreseeable future. Therapies under development and evaluation for patients with HCV infection include adjunctive use of the antiviral agent amantadine and the immunomodulatory agent thymalfasin. Novel small molecules include the ribavirin analogues, viramidine and levovirin, and BILN 2061, an inhibitor of HCV serine protease. Other therapeutic strategies that have reached the clinic include antisense oligonucleotides (ISIS 14803), nuclease-resistant ribozymes targeting HCV RNA (Heptazyme), human monoclonal antibodie, and human antibody fragments directed at HCV helicase. PMID- 15346253 TI - [Modern imaging of the heart, MRI or MSCT?]. PMID- 15346254 TI - [Micro-computed tomography of the vasculature in parenchymal organs and lung alveoli]. AB - Micro-CT has become a powerful technique in non-destructive 3D imaging and morphometric analysis. First results were limited to the investigation of osteoporosis in cancellous bone. But the availability of systems with almost microscopic resolution and sufficient soft tissue contrast has opened up entirely new applications for laboratory investigation of blood vessels and soft tissues. This article gives an overview of micro-CT technology and the potential of three dimensional imaging of the vessel wall and soft-tissue architecture imaging in different organs using different contrast perfusion and staining techniques. Micro-CT provides quantitative information on human plaque morphology equivalent to histomorphometric analysis. Based on differences in grey-scale attenuations, micro-CT also correctly identifies atherosclerotic lesions that are histologically classified as fibrous plaques, calcified lesions, fibroatheroma, and lipid rich lesions. Micro-CT is a promising method to visualize the architecture of the renal vasculature and, importantly, to separate cortex and medulla for the visualization of glomeruli and their afferent and efferent arterioles. Micro-CT can determine the vascular surface in a defined placental volume. Combining of micro-CT data and total placental volume enables an estimation of the approximate surface of the placental vasculature. The diameter of opacified vessels in the investigated samples ranged from 2 mm (chorion plate artery) to 14 micro m (smallest vessel diameter, terminal loop). Recognizing that lung parenchyma can only be visualized if the alveoli are completely expanded and the contrast of the thin alveolar walls is enhanced, we tested two preparation methods: (1) fixation of lung tissue with formalin vapour and staining with silver nitrate, and (2) intravenous injection of a barium sulfate-gelatine-thymol mixture in vivo in the anesthetized animal. We evaluated the ability of this mixture to enter the pulmonary microcirculation and the technical feasibility of micro-CT to assess lung micro-architecture. PMID- 15346255 TI - Analysis of mice tumor models using dynamic MRI data and a dedicated software platform*. AB - PURPOSE: To implement a software platform (DynaVision) dedicated to analyze data from functional imaging of tumors with different mathematical approaches, and to test the software platform in pancreatic carcinoma xenografts in mice with severe combined immunodeficiency disease (SCID). MATERIALS AND METHODS: A software program was developed for extraction and visualization of tissue perfusion parameters from dynamic contrast-enhanced images. This includes regional parameter calculation from enhancement curves, parametric images (e. g., blood flow), animation, 3D visualization, two-compartment modeling, a mode for comparing different datasets (e. g., therapy monitoring), and motion correction. We analyzed xenograft tumors from two pancreatic carcinoma cell lines (BxPC3 and ASPC1) implanted in 14 SCID mice after injection of Gd-DTPA into the tail vein. These data were correlated with histopathological findings. RESULTS: Image analysis was completed in approximately 15 minutes per data set. The possibility of drawing and editing ROIs within the whole data set makes it easy to obtain quantitative data from the intensity-time curves. In one animal, motion artifacts reduced the image quality to a greater extent but data analysis was still possible after motion correction. Dynamic MRI of mice tumor models revealed a highly heterogeneous distribution of the contrast-enhancement curves and derived parameters, which correlated with differences in histopathology. ASPC1 tumors showed a more hypervascular type of curves with faster and higher signal enhancement rate (wash-in) and a faster signal decrease (wash-out). BXPC3 tumors showed a more hypovascular type with slower wash-in and wash-out. This correlated with the biological properties of the tumors. CONCLUSION: With the described software, it was possible to analyze tissue perfusion parameters in small xenograft tumor models in mice. Our data correlated with histopathological data, and the qualitative and quantitative perfusion parameters could distinguish two tumor entities with different growth characteristics. PMID- 15346256 TI - Feasibility of MR-guided angioplasty of femoral artery stenoses using real-time imaging and intraarterial contrast-enhanced MR angiography. AB - PURPOSE: To show the feasibility of magnetic resonance (MR) for guided interventional therapy of femoral and popliteal artery stenoses with commercially available materials supported by MR real-time imaging and intraarterial MR angiography. MATERIALS AND METHODS: Three patients (1 female, 2male), suffering from symptomatic arterial occlusive disease with stenoses of the femoral (n = 2) or popliteal (n = 1) arteries were included. Intraarterial digital subtraction angiography was performed in each patient pre- and post-interventionally as standard of reference to quantify stenoses. The degree of the stenoses reached from 71 - 88 %. The MR images were acquired on a 1.5 T MR scanner (Magnetom Sonata; Siemens, Erlangen, Germany). For MR-angiography, a Flash 3D sequence was utilized following injection of 5 mL diluted gadodiamide (Omniscan; Amersham Buchler, Braunschweig, Germany) via the arterial access. Two maximum intensity projections (MIP) were used as road maps and localizer for the interactive positioning of a continuously running 2D-FLASH sequence with a temporal solution of 2 images per second. During the intervention, an MR compatible monitor provided the image display inside the scanner room. Safety guidelines were followed during imaging in the presence of a conductive guidewire. The lesion was crossed by a commercially available balloon catheter (Wanda, Boston Scientific; Ratingen, Germany), which was mounted on a 0.035" guidewire (Terumo; Leuven, Belgium). The visibility was provided by radiopaque markers embedded in the balloon and was improved by injection of 1 mL gadodiamide into the balloon. After dilation, the result was checked by intraarterial MR angiography and catheter angiography. RESULTS: The stenoses could be correctly localized by intraarterial MR angiography. There was complete correlation between intraarterial MR angiography and digital subtraction angiography. The combination of guidewire and balloon was visible and the balloon was placed correctly to cover the entire stenoses. Balloon dilation reduced the degree of stenosis about 45 % on average. A prolonged dilation was necessary in one patient due to a relevant residual stenosis, which was recognized by intraarterial MR angiography. CONCLUSION: MR guided balloon dilatation of femoral and popliteal artery stenoses supported by real time imaging and intraarterial MR angiography is feasible with commercially available materials in the above mentioned way. PMID- 15346257 TI - [Evaluation of regional wall motion abnormalities of the heart--comparison with doppler tissue echocardiography, MR-tagging and levocardiography]. AB - PURPOSE: To compare the visual analysis of magnetic resonance imaging (MRI) with the tagging technique and Doppler tissue echocardiography with invasive ventriculography in detecting and quantifying regional left ventricular wall motion abnormalities. MATERIALS AND METHODS: Sixteen patients with coronary artery disease and a history of prior myocardial infarction underwent invasive ventriculography, Doppler tissue echocardiography and MR-tagging within one week. Regional wall motion abnormalities (WMA) were detected in all patients. WMA were graded as normal = 1; hypokinetic = 2; akinetic = 3; or dyskinetic = 4. For agreement between MRI, echocardiography, and ventriculography the kappa coefficient (kappa) according to Cohen was calculated. RESULTS: The kappa coefficient (kappa) was 0.962 for agreement between MRI and echocardiography and 0.602 for agreement between MRI and ventriculography as well as between echocardiography and ventriculography. CONCLUSION: Reliable analysis of regional left ventricular wall motion abnormalities is feasible using visual analysis of MR-tagging. MRI and Doppler tissue echocardiography detect more WMA than invasive ventriculography and grade them as more severe. PMID- 15346258 TI - [Anthracycline-induced cardiotoxicity: cardiac MRI after treatment for childhood cancer]. AB - PURPOSE: Quantification of left and right ventricular function using MRI in young cancer patients treated with cardiotoxic anthracyclines. MATERIALS AND METHODS: Twenty-eight patients (mean age 16.4 years) underwent cardiac MRI at 1.5 T. The study protocol consisted of morphologic T2-weighted images with fat suppression and cine steady-state free precession sequences (SSFP) for functional analysis. Seven patients were examined at the end of chemotherapy, two of them also repeatedly during therapy, and 21 patients following an average period of three years after finishing chemotherapy (range one month--20 years) RESULTS: The end systolic volume index increased and the ejection fraction of the left and right ventricle decreased during anthracycline therapy. Two of seven patients showed a myocardial edema at the end of the therapy. In 15 of all 28 patients, the left ventricular ejection fraction was reduced to less than 55 % (minimum 44 %). No clinical signs of cardiac insufficiency or cardiomyopathy were observed. CONCLUSION: MRI is able to detect acute as well as chronic subclinical cardiotoxic effects of anthracyclines. Impairment of the right ventricular function should be considered in the diagnosis of anthracycline-induced cardiomyopathy. PMID- 15346259 TI - [Ultrasound follow-up of palpable solid, probably benign breast lesions (BI-RADS category III)]. AB - PURPOSE: To determine whether palpable non-calcified solid breast masses with benign morphology on ultrasound can be classified as probably benign and whether these lesions fulfill the criteria required for BI-RADS (TM) category III. MATERIALS AND METHODS: This analysis includes 78 patients with 80 palpable non calcified solid masses that were classified as probably benign at initial sonography. Of 78 patients, 66 had periodic imaging surveillance (2 - 8 years, mean 4.6 years). The remaining 12 patients had a biopsy after initial imaging. RESULTS: In the 64 patients under surveillance, palpable lesions remained stable at follow-up. In 2 patients lesions increased in size during follow-up and no malignancy was diagnosed at subsequent open biopsy. In 19 of the 66 patients, 29 additional nonpalpable lesions of similar morphology were detected on ultrasound. No malignancy was diagnosed in the 12 patients, who had a biopsy after initial imaging. CONCLUSION: Our data suggest that palpable non-calcified solid breast masses with benign morphology on ultrasound can be classified as probably benign (BI-RADS (TM) category III). Biopsy can be averted when lesions remain stable at follow-up. PMID- 15346260 TI - [Computed tomography (CT) of acute diverticulitis of the cecum and ascending colon]. AB - Acute diverticulitis of the cecum and ascending colon, also called right-sided diverticulitis, represents a relatively rare disorder in the western hemisphere. Pseudodiverticula and, less frequently, solitary congenital diverticula are regarded as the underlying causes of acute diverticulitis. We report the helical CT findings in four patients with acute right-sided colonic diverticulitis. The CT was performed with a collimation of 8 mm, a pitch of 1.5 and an increment of 8 mm, and with variable administration of intravenous, oral and rectal contrast material. In two of the four patients, the acute diverticulitis was detected in the cecum and ascending colon, respectively. In two patients, the diagnosis could be confirmed during surgery and subsequent histologic examination of the resected specimen. On the initial CT studies, acute diverticulitis was correctly diagnosed in two patients and suspected in one patient without identifying an inflamed diverticulum. In one patient, the offending diverticulum in the ascending colon caused an inflammatory pseudotumor at the level of the ileocecal region. This process was initially mistaken as Crohn's disease. The CT diagnosis of a right sided colonic diverticulitis is based on an inflamed diverticulum in the center of pericolic inflammatory changes and a preserved wall enhancement (target sign). Other CT findings, such as fatty pericolic infiltration and colon wall thickening, are rather non-specific and can also be found in a number of different ileocolic disorders, especially in colon cancer. In selected cases, the diagnosis can only be established by follow up CT after the pericolic infiltration has markedly subsided and an offending diverticulum has emerged. PMID- 15346261 TI - [Value of pelvic MRI in the preoperative diagnosis of endometriosis]. AB - PURPOSE: To determine the value of magnetic resonance imaging (MRI) of the pelvis in the preoperative diagnosis of endometriosis. MATERIALS AND METHODS: Over a period of 8 months, preoperative MRI of the pelvis were obtained in 13 patients with suspected endometriosis (mean patient age 34.6 years; range 25 - 47 years). RESULTS: In 9 of 13 patients (69 %), the diagnosis of endometriosis was made by MRI and confirmed by laparoscopy in 8 cases. In 2 of 13 patients, endometriotic lesions were detectable by laparoscopy only. In the remaining 2 patients, no endometriosis was visible on MRI or by laparoscopy. MRI was able to visualize a total of 19 endometriotic lesions, with 14 (74 %) confirmed by histopathologic examination following laparoscopy. Five of these 19 lesions (26 %) visible on MRI were not seen by laparoscopy. Using laparoscopy and subsequent histopathologic examination, 27 endometriotic lesions were diagnosed, with 13 (48 %) not seen on the preoperative MRI. CONCLUSION: MRI and laparoscopy are complementary diagnostic tools that will best document the full extent of endometriosis when combined. MRI can visualize additional lesions inaccessible to laparoscopy. Thus, MRI of the pelvis should used preoperatively for surgical treatment planning. PMID- 15346262 TI - [Diagnosis and classification of calcaneal fractures in computed tomography]. AB - PURPOSE: To compare image quality of single-slice spiral CT (SSCT) and multislice spiral CT (MSCT) in the diagnosis and classification of calcaneal fractures and to present a rapid and precise algorithm for the classification system of Stuermer. MATERIALS AND METHODS: In 102 patients with 124 calcaneal fractures, spiral CT was performed, in 82 cases as SSCT with a slice thickness (SD) of 3 mm, a table speed (TS) of 3 - 4 mm/rot and an increment of 1.5 mm. In 42 cases, patients were scanned using MSCT (SD of 1.25 mm, increment 0.8 mm). For these examinations, 2 different scan protocols were used, with a TS of 3.75 mm/rot in one group (n = 21) and a TS of 7.5 mm/rot in the other group (n = 21). The image quality of axial sections and reconstructed images was assessed on a scale from 1 to 5 (1 = very good; 5 = insufficient). The fractures were evaluated using a classification system according to Stuermer, which assigns three main groups (A/B/C) and three subtypes (1/2/3). RESULTS: MSCT had substantial advantages over SSCT with respect to scan time and image quality, especially for multiplanar reformatting (median 1.5 versus 4.0). TS showed no significant influence on the image quality. The standardized evaluation of the images enabled a classification of fractures within 5 minutes. All fractures could be assigned to the different types and subtypes. This classification system takes into account the severity of the fracture and the therapeutic approach. The most common type (90 of 124 fractures) were "joint depression" fractures (type C), which were treated by surgery in 92 %. Type A fractures were treated conservatively in 72 %. CONCLUSION: Spiral CT, especially MSCT, allows rapid diagnosis and precise classification of calcaneal fractures, achieved with high quality multiplanar reformatting. The presented classification in different fracture types and subtypes allows an adequate planning of therapy. PMID- 15346263 TI - [Evaluation of tibial torsion--comparison of clinical methods and computed tomography]. AB - PURPOSE: To determine variability between clinical goniometric methods and computed tomography (CT) in measuring posttraumatic malrotation of the tibia. MATERIALS AND METHODS: In a prospective study, absolute tibial torsion of both legs after unilateral fracture of the tibia as well as the difference between both legs (intra-individual torsional difference) was postoperatively determined with two goniometric and two CT methods in 40 patients (female : male = 16 : 24, mean age = 46 +/- 34 years). RESULTS: The mean difference between goniometric and CT methods in determining intra-individual torsional difference was not significant. Nevertheless, variance of values was higher in goniometric measurement (up to +/- 11 degree), with the measurements of torsional difference showing two times greater standard deviation in interobserver variability of goniometric methods as compared to CT. CONCLUSION: Goniometric methods do not significantly differ from the CT methods for determination of intraindividual torsional difference. The higher variance of interobserver values limits accuracy of goniometric methods. Therefore, goniometric determination of tibial torsion can only be considered an estimate but not a precise measurement. PMID- 15346264 TI - [Dynamic MRI of the bone marrow for monitoring multiple myeloma during treatment with thalidomide as monotherapy or in combination with CED chemotherapy]. AB - PURPOSE: To quantify changes of bone marrow microcirculation in multiple myeloma (MM) using contrast enhanced dynamic MRI (dMRI) during thalidomide as antiangiogenic monotherapy or in combination with chemotherapy (cyclophosphamide, etoposide, dexamethasone). MATERIALS AND METHODS: The study includes 63 patients with refractory or relapsed MM, who underwent dMRI with high temporal resolution (T1w-turboFLASH) of the lumbar spine before and following treatment. The contrast uptake was quantified using a two compartment model with the output parameters amplitude and k (ep) (exchange rate constant). The evaluation considered the initial dMRI finding (pathological or non-pathological) and the clinical therapeutic response (response or no response). RESULTS: During monotherapy with thalidomide (n = 38), no significant changes of the dMRI parameters were found, even when considering the initial dMRI finding (positive n = 22) and the therapeutic response (responder n = 14). The combination with chemotherapy (n = 25) had a significant reduction of k (ep) (p = 0.01) in 18 patients with positive initial dMRI finding and therapeutic response. Reduction of the amplitude was seen in most cases, but in the end without any significance (p = 0.09). CONCLUSION: dMRI can quantify significant changes of bone marrow microcirculation solely during treatment with thalidomide combined with chemotherapy, not with thalidomide alone. PMID- 15346265 TI - [Influence of sequence type on the extent of the susceptibility artifact in MRI- a shoulder specimen study after suture anchor repair]. AB - PURPOSE: To compare the extent of susceptibility artifacts after metallic suture anchor implantation by analyzing 14 different MRI sequences. MATERIALS AND METHODS: A metallic suture anchor was implanted in the central area of three glenoid porcine specimens. The specimens were imaged with a 1.5 T scanner using a protocol of 14 standard sequences including gradient echo, spin echo and turbo spin echo sequences with and without fat-saturation. Artifact size was measured for each specimen and sequence. The resulting mean artifact areas were determined for each type of sequence and the mean values of the three specimens compared. RESULTS: Gradient echo-sequences produced significantly larger artifact areas than the spin echo and turbo spin echo sequences, whereby the artifacts of the 3D gradient echo sequences were smaller than the artifacts of the 2D-gradient echo sequences. A turbo spin echo sequence with a high readout band width and a short effective echo time showed the best results. For the conventional spin echo sequence, a reduction in the echo time did not significantly decrease the artifact size. Spectral fat-saturation did not affect the area of the susceptibility artifact compared to the non-saturated sequence. CONCLUSION: Gradient echo sequences should not be used after metallic suture anchor repair. Turbo spin echo sequences showed a decrease in the artifact size compared to conventional spin echo sequences and should be performed with a short effective echo time and a high band width. Spectral fat- saturation did not increase the artifact size significantly. PMID- 15346266 TI - [PTA versus Palmaz stent placement in femoropopliteal artery stenoses: results of a multicenter prospective randomized study (REFSA)]. AB - PURPOSE: To evaluate whether stent placement is superior to percutaneous transluminal angioplasty (PTA) in the treatment of chronic symptoms in short femoropopliteal arterial stenoses. MATERIALS AND METHODS: One hundred twenty-four limbs in 116 patients, who ranged in age from 39 to 87 years (mean age, 67 years), were randomized to PTA (n = 53) versus PTA followed by implantation of long-medium Palmaz-Stents (n = 71). Inclusion criteria were intermittent claudication or chronic critical limb ischemia, short stenosis or occlusion (lesion length < or = 5 cm), and at least one patent run-off vessel at angiography. The follow-up included clinical assessment, measurement of ankle/brachial index (ABI), color duplex ultrasound, and/or angiography at 6, 12 and 24 months. Angiographic follow-up between 12 and 36 months was available in 54 limbs (45 %). RESULTS: Initial technical success was achieved in 50 of 53 limbs (94.4 %) in the PTA group versus 70 of 71 limbs (98.6 %) in the stent group. Overall, major complications occurred in 9.5 % (n = 11), with n = 4 in the PTA group compared to n = 7 in the stent group. No difference was found between the groups of treatment: clinical success at 1 and 2 years was 80.5 and 77.1 % in the PTA group versus 78.1 and 71.0 % in the stent group. The cumulative 1-year and 2-year angiographic primary patency rates were 66.1 and 49.1 % in the stent group versus 76.1 and 66.1 % in the PTA group. The secondary 1-year and 2-year angiographic patency rates were 88.5 and 53.3 % in the stent group versus 82.7 % and 76.2 % in the PTA group. CONCLUSION: The primary success rate was slightly higher after stent placement than after PTA. However, the angiographic, clinical and hemodynamic success after 1 and 2 years tends to be slightly better for PTA. PMID- 15346267 TI - [In vitro examination of the visibility of 11 stent catheters with real-time MR imaging]. AB - PURPOSE: To evaluate artifacts of unexpended stents and to determine their exact position for MR-guided percutaneous transluminal angioplasty using real-time sequences. MATERIALS AND METHODS: By using an in vitro model, 11 unexpended stents - 9 nitinol, 1 stainless steel, and 1 cobalt alloy - were investigated by MR. Each stent was studied in a vessel-phantom filled with saline solution. Imaging was performed using five different real-time sequences: fast low angle shot (Flash 2D), fast imaging with steady precession (true FISP, FISP, interactive true FISP) and segmented echo planar imaging (seg. EPI). Artifacts of the introducer system and the stent were calculated by four blinded radiologists (scale: 1 - artifacts, making an excellent contribution to visualization; 2 - artifacts, making mainly a contribution to visualization; 3 - artifacts, making no contribution to visualization). Furthermore, an evaluation of the visibility of the tip of the stent-catheter and the proximal and distal end of the stent was performed using a four-point scale (very good visibility to invisible). RESULTS: The artifacts of the introducer system and stent were rated best for Omnilink (1.3 +/- 0.47), Wallstent (1.6 +/- 0.5), Jostent (1.65 +/- 0.5) and Luminexx (1.65 +/- 0.5). The differences between Omnilink and Jostent as well as Omnilink and Luminexx were significant. A very good to good visibility of the catheter tip was observed with a mean of 1.7 +/- 0.66 for Omnilink followed by the Jostent (1.95 +/- 0.69), by the Wallstent(R) (2.1 +/- 0.72) and by Luminexx (2.5 +/- 1.14). Differences between Omnilink and Luminexx were significant. The visibility of the proximal and distal end of the stent was evaluated as very good to good in 4 stent catheters (Omnilink, Wallstent, Jostent, Luminexx). However, the differences between Omnilink and Jostent as well as between Omnilink and Luminexx, and between Wallstent and Luminexx were significant. For all evaluation criteria, the segmented EPI and the interactive true FISP were rated to be most suitable for visualization. There were no significant differences in the evaluation of these two sequences. CONCLUSION: Omnilink stent, Jostent, Wallstent, and Luminexx stent are suitable for MR-angioplasty due to their good visibility in MR. Segmented EPI- and a newly developed interactive true FISP sequence enable an effective visualization of these stent-catheters. PMID- 15346268 TI - [Reimbursement of radiologically guided vascular interventions within the DRG system: what will change?]. AB - PURPOSE: To evaluate reimbursement within the DRG-system ("diagnosis-related groups") compared with traditional reimbursement for interventional therapy of hospitalized patients. MATERIALS AND METHODS: Reimbursement calculation was prospectively analyzed in two respects for 30 consecutive patients who underwent percutaneous transluminal angioplasty (PTA) of the lower extremity arteries: (1) based on the DRG-system; (2) based on the traditional system. Additional evaluation was performed for five further, typical vascular procedures on the basis of real documentation and calculation data (stenting of the carotid artery, fibrinolytic therapy of basilar artery occlusion, stenting of renal artery stenosis, angioplasty of hemodialysis-shunt stenosis and aspiration thrombectomy of an infrapopliteal arterial occlusion). RESULTS: In our hospital, the introduction of the DRG system would reduce reimbursement by approximately 1100 euro per PTA patient. However, the other vascular radiological procedures can be expected to increase the payments by up to 4500 euro. CONCLUSION: To minimize imminent reduction of reimbursement for patients with peripheral PTA, complete documentation and economical patient management is mandatory. Payment may increase significantly for patients with the other reported vascular interventional procedures. PMID- 15346269 TI - [Cyclosporin A-induced posterior reversible encephalopathy syndrome (PRES) in MRI]. PMID- 15346270 TI - [Selective catheter embolization in idiopathic refractory epistaxis]. PMID- 15346271 TI - [Rectal manifestation of B-cell non-Hodgkin's lymphoma. A rare cause of anorectal pain]. PMID- 15346272 TI - [Silicone granuloma in intrathoracic lymph nodes after rupture of a silicone double-chamber prosthesis]. PMID- 15346273 TI - [Granular Echinococcus]. PMID- 15346282 TI - [Non-invasive assessment of coronary artery bypass grafts -- an update]. AB - The limited lifetime and the correlation between graft occlusion and recurring symptoms underline the need for repeated imaging of coronary artery bypass grafts. CT and MRI allow for non-invasive imaging of coronary bypasses with high accuracies concerning the patency of these vessels. Multidetector CT seems to be the CT technique of choice, especially after the introduction of 16 slice CT scanners for morphologic assessment of coronary artery bypass grafts. Compared with MRI, CT is a robust technique for assessment of cardiac anastomoses, native coronary arteries, and for the detection of graft stenoses. MRI, however, is able to deliver functional information about the grafts and the recipient coronary arteries by determining the coronary flow reserve. Furthermore, it can be integrated in a multiparametric MR examination protocol. The follow-up of asymptomatic patients can primarily be done by these non-invasive techniques as nearly every third patient reveals an asymptomatic bypass occlusion 5 years after operation. Furthermore, patients with atypical complaints after the operation may undergo non-invasive imaging as long as documented patency of the bypass averts coronary angiography. Patients with recurrent angina pectoris and/or myocardial ischemia discovered by other cardiologic tests have to undergo coronary angiography. PMID- 15346283 TI - [Radiation exposure and image sharpness of mammographic units in bavaria with respect to the European guidelines]. AB - In a survey including all mammography units in Bavaria, entrance doses in air and other quality-relevant parameters were measured with standard settings and under comparable conditions. The study was aimed to discover discrepancies between practical mammographic imaging techniques and the demands established in the European Guidelines. Test images were made by the operators themselves with phantoms used for constancy checks. Entrance dose values were measured with official film dosimeters. The survey showed that more than 95 % of all mammographic units in Bavaria comply with the European guidelines with respect to radiation exposure and imaging sharpness. PMID- 15346284 TI - MR elastography of the prostate: initial in-vivo application. AB - PURPOSE: To analyze the initial assessment of the technical feasibility of in vivo MR elastography (MRE) of the prostate gland in healthy volunteers. MATERIALS AND METHODS: Dynamic sinusoidal MR elastography was performed in 7 healthy volunteers in prone position. The mechanical wave was induced via an external oscillator attached to the pubic bone. A 1.5 Tesla MR system (Philips Medical Systems, Netherland) was used with 4 combined surface coils for signal reception. MRE data acquisition was performed with a motion-sensitive spin-echo MR sequence that was phase-locked to the mechanical oscillation. Subsequently, these images were used to reconstruct the local distribution of elasticity inside the prostate gland. The applied reconstruction algorithm was tested by means of phantom measurements. RESULTS: Sufficient penetration of the mechanical wave into the prostate gland was achieved in all volunteers, allowing the acquisition of utilizable image data sets. The reconstructed distribution of elasticity (shear modulus) inside the healthy prostate gland correlated with the zonal anatomy of the gland. The elasticity of the central portion (2.2 +/- 0.3 kPa) appeared to be lower than the peripheral prostatic portion (3.3 +/- 0.5 kPa). CONCLUSION: In vivo MRE of the prostate gland is technically feasible. The proposed experimental set-up allows the efficient insertion of the mechanical wave into the prostate gland and provides a successful MR data acquisition. PMID- 15346285 TI - Two-dimensional parallel acquisition technique in 3D MR colonography. AB - PURPOSE: In-vitro and in-vivo examinations to investigate the influence of one (1D)- and two-dimensional (2D) parallel acquisition techniques (PAT) on contrast to-noise ratio (CNR) and image quality for bright-lumen 3D MR colonography. MATERIALS AND METHODS: In-vitro measurements were performed on a 1.5 T whole-body MR scanner (MAGNETOM AVANTO(R), Siemens AG, Erlangen) with a standard spoiled 3D gradient-echo (3D GRE) sequence and a volume interpolated 3D GRE (VIBE) sequence using a home-built colon phantom. The relative CNR was determined and image quality evaluated for different acceleration factors (PAT factors). A bright lumen 3D MR colonography with PAT factors 2, 3, 4 and 6 was performed on a 39 year-old volunteer. The 3D data sets were compared with regard to image quality. RESULTS: The mean relative CNR values for the in-vitro measurements were 1, 0.81, 0.73, 0.52 and 0.4 (3D GRE) and 1, 0.8, 0.65, 0.45 and 0.3 (VIBE) for the PAT factors 1, 2, 3, 4 and 6, respectively. Residual aliasing artifacts not affecting image quality in a distinct manner were visible for 2D PAT, especially for the VIBE sequence. Increasing the PAT factor up to 6 and decreasing the acquisition time down to 10 seconds for the bright-lumen 3D MR colonography could achieve adequate image quality with significantly reduced image artifacts caused by peristalsis and pulsations. DISCUSSION: Even for high PAT factors up to 6, 2D PAT only leads to a moderate CNR loss. For a 3D MR colonography, distinct shorter acquisition times can be achieved with identical resolution. PMID- 15346286 TI - [Virtual MR endoscopy of the ventricles prior to neurosurgical interventional endoscopy -- evaluation of different presentation techniques]. AB - PURPOSE: In the past, virtual endoscopies have been performed for planning of endoscopic interventions or for diagnostic purposes in various organ systems with increasing frequency. This study evaluates the ability of virtual ventricular endoscopy to depict anatomical structures and the use for planning of real endoscopy. MATERIALS AND METHODS: In a prospective study, 4 volunteers and 8 patients were examined with MRI. In 3 of the patients endoscopy was performed by our neurosurgeons thereafter. The calculation of the virtual endoscopy was based on 1 mm sagittal T2-weighted images. Comparison of surface rendering and volume rendering was made by means of video sequencing of individual views, and these were compared with the intraoperative endoscopic videos concerning the depictability of anatomical landmarks. RESULTS: The reconstructions using volume rendering were more significant and easier to calculate than those based on surface rendering. Virtual endoscopy in the transparent mode allowed visualization of hazardous structures outside the ventricular system such as the basilar artery tip. Transparent 3D images of the ventricles gave a good overview on the depicted structures and enabled a better orientation during the virtual camera flight than surface rendered views. CONCLUSION: MR-based virtual endoscopy of the ventricular system can be obtained on the basis of surface- and volume rendered views of sagittal T2-weighted thin sections. Preoperative utilization of this method simplifies the planning of endoscopy by visualization of anatomical structures. PMID- 15346287 TI - [Metabolic imaging to follow stereotactic radiation of gliomas -- the role of 1H MR spectroscopy in comparison to FDG-PET and IMT-SPECT]. AB - PURPOSE: To evaluate the clinical value of 1H MR spectroscopy (1H MRSI) for follow-up of irradiated glioma compared to positron emission tomography (PET) with [18F]-2-fluoro-deoxy-D-glucose (FDG-PET) and single photon emission tomography with [123I]-a-methyl-L-tyrosine (IMT-SPECT). MATERIALS AND METHODS: Twenty-four patients with irradiated gliomas were examined using 1H MRSI (2D spectroscopic imaging; PRESS; TE = 135 msec; 1.5T Magnetom Vision, Siemens; Voxel size 9 x 9 x 15 mm (3)). MR spectra (n = 233) were evaluated in areas suspicious of tumor (n = 86) as well as in healthy appearing brain tissue (n = 147). Relative signal intensity ratios of choline (Cho), creatine (Cr) and N-acetyl aspartate (NAA) were calculated. PET scans (n = 19) were performed with 200 - 250 MBq FDG, IMT-SPECT examinations (n = 14) with 200 - 250 mBq IMT. Based on clinical and MRI/CT, follow-up lesions were classified as either neoplastic [PT] or non-neoplastic [nPT]. RESULTS: True positive results for the diagnosis of PT/nPT were 88/89 % (1H MRSI), 73/100 % (PET) and 100/75 % (SPECT). Cho/Cr showed highly significant changes for PT. Determinating a correlation between Cho, Cr, NAA and IMT-SPECT as well as FDG-PET was not possible because of different location of maximum tracer uptake and acquired 2D 1H MRSI. CONCLUSION: IMT-SPECT seems to be superior to detect tumor progression in irradiated gliomas. 1H MRSI was more suitable than FDG-PET to differentiate between recurrence and radiation induced changes. FDG-PET plays a role as sensitive method for detecting high grade tumors. PET and SPECT allowed the examination of the entire tumor including surrounding brain tissue with higher spatial resolution than the acquired 2D 1H MRSI. A main limitation of our study was that only 2D 1H MRSI was used, with only parts of the tumor evaluated. The use of 3D MR spectroscopic imaging may further increase the diagnostic accuracy. PMID- 15346288 TI - [Indications of HR-CT in the early postoperative phase of stapedotomy]. AB - PURPOSE: This is an analysis of high-resolution computed tomography (HR-CT) performed in patients with and without symptoms after stapes surgery during the early postoperative phase. MATERIALS AND METHODS: We examined 37 patients who underwent stapes surgery between January 1995 and December 2000. HR-CT of the temporal bone was performed in axial and coronal planes with 1-mm-slice thickness and table feed. A reoperation was performed in 7 patients. CT findings were compared with clinical follow up and surgical findings. RESULTS: All patients showed postoperative opacification of the tympanic cavity and external auditory canal as well as partial opacification of the mastoid on CT scans. A dislocation of the prosthesis was found in 11 cases and air bubbles were seen in the vestibule of 5 patients. In one case, small bone fragments were detected in the vestibule. In 6 of 7 cases, CT results were confirmed by retympanotomy. CONCLUSIONS: Due to overlapping of normal and pathological findings in the early postoperative period, indications for HR-CT are limited in this period. PMID- 15346289 TI - [Phantom study for the detection of simulated lesions in five different digital and one conventional mammography system]. AB - PURPOSE: Experimental phantom study for the detection of simulated lesions with five different digital and one conventional screen-film mammography system. MATERIALS AND METHODS: Three radiographs were obtained at various configurations of the phantom with one conventional screen-film system (Mammomat 3000 N) (Siemens), five digital systems (high resolution computed radiography system [Fuji/Siemens], one a-Si detector [GE Medical Systems], two a-Se detectors [Siemens; Hologic / Lorad] and one CCD detector [Fischer Imaging]), applying the same exposure parameters. The Wisconsin Mammographic Random Phantom, Model 152 A, was used. Five investigators with different experience in mammography (3 months to more than 4 years) evaluated the 18 randomly selected radiographs. RESULTS: No significant differences were found in the detection rate of simulated breast lesions for conventional screen-film mammography (84.9 %), high resolution computed radiography (86.7 %) and digital mammography with an a-Si detector (89.8 %). Highly significantly better results (p < 0.001) were found with the two a-Se systems (97.3 %) and the CCD system (100 %). CONCLUSION: Conventional screen-film mammography can be replaced by high resolution computed radiography and digital mammography with a-Si-, a-Se- and CCD-detectors. This has to be confirmed in further clinical studies. PMID- 15346290 TI - [The value of applying nitroglycerin in 3D coronary MR angiographie with real time navigation technique]. AB - PURPOSE: Nitroglycerin administration results in dilation of epicardial coronary vessels and in an increase in coronary blood flow, and has been suggested to improve MR coronary angiography. This study evaluates systematically whether administration of nitroglycerin improves the visualization of coronary arteries and, as a result, the detection of coronary artery stenosis during free breathing 3D coronary MR angiography. MATERIALS AND METHODS: Coronary MR angiography was performed in 44 patients with suspected coronary artery disease at a 1.5 Tesla System (Intera, Philips Medical Systems) (a) with and (b) without continuous administration of intravenous nitroglycerin at a dose rate of 2.5 mg/h, using an ECG gated gradient echo sequence with real-time navigator correction (turbo field echo, in-plane resolution 0.70 x 0.79 mm(2), acquisition window 80 ms). Equivalent segments of the coronary arteries in the sequences with and without nitroglycerin were evaluated for visualized vessel length and diameter, qualitative assessment of visualization using a four point grading scale and detection of stenoses > 50 %. Catheter coronary angiography was used as a gold standard. RESULTS: No significant differences were found between scans with and without nitroglycerin as to average length of the contiguously visualized vessel length (p > 0.05) and diameter (p > 0.05). There was also no significant difference in the coronary MR angiography with and without nitroglycerin in the average qualitative assessment score of the visualization of LM, proximal LAD, proximal CX, and proximal and distal RCA (2.1 +/- 0.8 and 2.2 +/- 0.7; p > 0.05). Sensitivity (77 % [17/22] vs. 82 % [18/22] p > 0.05) and specificity (72 % [13/18] vs. 72 % [13/18] p > 0.05) for the detection of coronary artery stenosis also did not differ significantly between scans with and without intravenous administration of nitroglycerin. CONCLUSION: Administration of nitroglycerin does not improve visualization of the coronary arteries and detection of coronary artery stenosis in free breathing 3D coronary MR angiography. PMID- 15346291 TI - [Computer simulation of a clinical magnet resonance tomography scanner for training purposes]. AB - PURPOSE: The idea for this project was born by the necessity to offer medical students an easy approach to the theoretical basics of magnetic resonance imaging. The aim was to simulate the features and functions of such a scanner on a commercially available computer by means of a computer program. MATERIALS AND METHODS: The simulation was programmed in pure Java under the GNU General Public License and is freely available for a commercially available computer with Windows, Macintosh or Linux operating system. The graphic user interface is oriented to a real scanner. In an external program parameter, images for the proton density and the relaxation times T1 and T2 are calculated on the basis of clinical examinations. From this, the image calculation is carried out in the simulation program pixel by pixel on the basis of a pulse sequence chosen and modified by the user. The images can be stored and printed. In addition, it is possible to display and modify k-space images. RESULTS: Seven classes of pulse sequences are implemented and up to 14 relevant sequence parameters, such as repetition time and echo time, can be altered. Aliasing and motion artifacts can be simulated. As the image calculation only takes a few seconds, interactive working is possible. CONCLUSION: The simulation has been used in the university education for more than 1 year, successfully illustrating the dependence of the MR images on the measuring parameters. This should facititate the approach of students to the understanding MR imaging in the future. PMID- 15346292 TI - [Bavarian mammography screening: results of consistency tests in X-ray units over three months]. AB - PURPOSE: To find out whether the quality requirements in daily, weekly or monthly tests of the "European protocol for the quality control of the physical and technical aspects of mammographic screening (EPQC)" can be maintained when screening the population of a large area. Furthermore, to check the specific tolerances of different test positions given by the EPQC for redundancy and statistical correlation. MATERIALS AND METHODS: Daily test images were obtained with an additional exposition of a sensitometric step wedge according DIN 6868 - 2. Weekly densitometry and visual inspection were performed at the University of Applied Sciences, Cologne. RESULTS: All units were able to maintain basic requirements on image quality as long as films of the same batch number were used. Some tolerance requirements of the EPQC are inconsistent and redundant. CONCLUSION: The occasionally considerable fluctuations of film processing require a daily check on the processing conditions. Daily test images of the units are only obligatory in the first 4 to 5 weeks of a new operation. All test results should be analyzed retrospectively in an independent test laboratory with emphasis on the statistical behavior of the unit. The tolerances prescribed by the EPQC are not always consistent and should be adapted to today's scientific standard when applied in mammography screening centers. PMID- 15346293 TI - [Development of a secure and cost-effective infrastructure for the access of arbitrary web-based image distribution systems]. AB - PURPOSE: To build an infrastructure that enables radiologists on-call and external users a teleradiological access to the HTML-based image distribution system inside the hospital via internet. In addition, no investment costs should arise on the user side and the image data should be sent renamed using cryptographic techniques. MATERIALS AND METHODS: A pure HTML-based system manages the image distribution inside the hospital, with an open source project extending this system through a secure gateway outside the firewall of the hospital. The gateway handles the communication between the external users and the HTML server within the network of the hospital. A second firewall is installed between the gateway and the external users and builds up a virtual private network (VPN). A connection between the gateway and the external user is only acknowledged if the computers involved authenticate each other via certificates and the external users authenticate via a multi-stage password system. All data are transferred encrypted. External users get only access to images that have been renamed to a pseudonym by means of automated processing before. RESULTS: With an ADSL internet access, external users achieve an image load frequency of 0.4 CT images per second. More than 90 % of the delay during image transfer results from security checks within the firewalls. Data passing the gateway induce no measurable delay. CONCLUSION: Project goals were realized by means of an infrastructure that works vendor independently with any HTML-based image distribution systems. The requirements of data security were realized using state-of-the-art web techniques. Adequate access and transfer speed lead to a widespread acceptance of the system on the part of external users. PMID- 15346294 TI - [Primary bilateral renal Burkitt lymphoma in childhood]. PMID- 15346295 TI - [Sonographic demonstration of a non-palpable Implanon implant using tissue harmonic imaging--three case reports]. PMID- 15346296 TI - [Use of multi-detector tier CT (16x) for angiographic intervention planning in pulmonary atrioventricular malformation]]. PMID- 15346297 TI - [Diagnosis of a neurofibroma, a small malformation associated with endometrial carcinoma in neurofibromatosis type 1 (Recklinghausen)]. PMID- 15346298 TI - [The prevalence of pain and deformities in the feet of adolescents. Results of a cross-sectional study]. AB - AIM: The purpose of the study was to evaluate the prevalence of foot pain and foot deformities in adolescents. It was aimed to obtain information on the association between foot pain and foot deformities. METHOD: A total of 2 368 adolescents (age 14.5 +/- 0.7 years) were evaluated. The frequency of foot pain was probed by using a self-reporting questionnaire. The foot deformities were evaluated during clinical examinations by school doctors. RESULTS: The prevalence of foot pain was 14.0 % and the prevalence of foot deformities was 13.7 %. The prevalence of pain was significantly higher in adolescents with foot deformity (17.8 %) than in persons without deformity (13.4 %), p < 0.05. The prevalence of a flexible flat foot was 6.2 % and the prevalence of a rigid flat foot was 0.5 %. Other deformities registered were splay foot (2.3 %) and flexible splay-flat foot (2.0). The prevalence of hallux-valgus deformity was 3.5 %. A total of 3.5 % patients were suffering from a plantar hyperkeratosis. This was significantly correlated to a high pain prevalence (the hyperkeratosis was significantly associated with a high prevalence of pain). Significant factors which were significantly associated with foot pain were foot deformity (1.4 fold) and hyperkeratosis (75 fold). Foot pain was 1.4 fold higher in children with foot deformity and 75 fold higher in feet with hyperkeratosis. CONCLUSION: The prevalence of foot pain and foot deformity in adolescent is high. Mild deformities (flexible flat foot and splay foot) are physiological variations without any association to foot pain. Pathological conditions that are associated with foot pain are the rigid flat foot, the hallux valgus and the cavus deformity. Plantar hyperkeratosis is an indicator of foot pathology. PMID- 15346299 TI - [Epiphyseal dysplasia--symptoms and differential diagnostic aspects]. AB - AIM: Children complaining of hip pain most likely suffer from Perthes' disease. Similar morphological changes are seen in the hereditary diseases of the epiphyseal dysplasias which have a prevalence of about 40 of 100 000 inhabitants. We now show the differentiation between bilateral Perthes' disease, epiphyseal dysplasia and skeletal disorders like mucopolysaccharidosis and pseudoachondroplasia. METHOD: We describe the diseases of multiple epiphyseal dysplasia and spondyloepiphyseal dysplasia on the basis of two case reports. Furthermore, we discuss differential diagnostic aspects, therapeutic options and prognosis. RESULTS: Epiphyseal dysplasias are classified as osteochondral dysplasia. The characteristic feature of these heredopathies is short statue due to skeletal dysplasias at various sites. Multiple epiphyseal dysplasia is caused by an abnormality of enchondral ossification. A distinction is made between the severe Fairbank form, the milder Ribbing form and a mild but rather localised Meyer form. In addition to the femoral head, the spinal bodies can also be affected by the same enchondral ossification disorders and this is defined as spondyloepiphyseal dysplasia. CONCLUSION: The diagnosis of a Perthes' disease can be made from an X-ray of the hip, showing either an advanced necrosis of the femoral head or early signs of it. Typical radiological findings allow the differentiation to other skeletal dysplasias like epiphyseal dysplasia and metabolic disorders. PMID- 15346300 TI - [Prospective evaluation of braces as treatment in idiopathic scoliosis]. AB - AIM: To prospectively evaluate the results of brace treatment in idiopathic scoliosis and to define risk factors of treatment failure. METHOD: Fifty-two patients with a Cobb angle of between 25 and 40 degrees were included in the study. Prior to initiation of brace treatment with the Cheneau-Toulouse-Muenster orthesis, skeletal age and flexibility of the curve (bending films) were evaluated. The average follow-up after weaning of the brace was 42 months (36-78 months). RESULTS: An average initial Cobb angle of 31 degrees was corrected to 18 degrees (43 %) under brace treatment with a flexibility to 6 degrees Cobb angle on bending films. Three years after weaning there was an overall increase of the Cobb angle to 37 degrees on average. The apical vertebral rotation was corrected from 16 degrees to 11 degrees (31 %) and increased to 20 degrees during follow up. Thoracic kyphosis changed from 24 degrees to 18 degrees during treatment. At the latest follow-up kyphosis had returned to the pre-treatment angle again. Twenty-two patients had a curve progression during or after brace treatment of more than 5 degrees. In 14 patients surgical correction and fusion have been indicated. There was a positive correlation between flexibility and Cobb angle correction during brace treatment and a negative correlation between Cobb angle correction during brace treatment and curve progression (p < 0.05). CONCLUSIONS: Curve progression was prevented in 58 %. Prognostic risk factors are a young age at initiation of brace treatment, a thoracic curve, unsatisfactory curve correction in the brace and a male gender. PMID- 15346301 TI - [Juxta-facet cysts as a differential diagnosis for lumbar neuralgia]. AB - AIM: The aim of the current investigation it was to evaluate the incidence, clinical symptoms and the results of surgical treatment of lumbar juxta-facet cysts. METHOD: Between January 2002 and July 2003 305 patients underwent decompression of the lumbar spinal canal. In 3.6 % of these (n = 11) juxta-facet cyst were found to be responsible for the complaints and resection of the cyst was performed. All patients underwent standardized clinical examination and pain evaluation by the means of a visual analogue scale in a follow-up of 6.8 month on average. RESULTS: The average history of lumbar pain was 26 months, that of leg pain 23 weeks, respectively. Radicular symptoms appeared in 7 patients, and 9 of 11 patients complained about spinal claudication. Clinical examination showed a typical pain provocation during reclination of the lumbar spine. However none showed a segmental instability. Preoperative MRI revealed facet cysts with an average diameter of 9 mm (4-18 mm). All of the patients showed signs of degenerative spondylarthritis. Postoperative examination revealed in 8 cases a very good and in 2 cases a good result. One patient complained about persisting sciatica in spite of MR-tomographic demonstration of complete resection of the cyst. The walking-distance improved by least 50 % in 10 cases. Although no spinal fusion was performed, no patient developed a postoperative segmental instability. CONCLUSION: Juxta-facet cysts represent a not uncommon differential diagnosis in patients with lumbar radiculopathy. In the case of missing signs of segmental instability, resection of the cyst without simultaneous spinal fusion seems to be an appropriate therapy. PMID- 15346302 TI - [BWS-mobility in patients with an impingement syndrome compared to healthy subjects--an inclinometric study]. AB - AIM: The aim of this study was to analyse the relation between the mobility of the thoracic spine and an impingement syndrome of the shoulder. METHOD: In a prospective study, 50 patients with an impingement syndrome and 50 healthy test subjects were examined for the mobility of their thoracic spines. All patients and test subjects were examined according to a standardized protocol. The experiments were carried out in the biomechanical laboratory of our clinic with the Plurimetercompass and the Inclinometer of Rippstein. RESULTS: In 23 patients a tendinosis calcarea was diagnosed radiologically, 27 patients suffered from a plain impingement without calcification, hence both groups were analyzed separately. The mobility of the thoracic spine in the sagittal and frontal planes and in rotation was significantly different between the three groups. The highest mobility was found in the healthy test subjects, the lowest in patients with a plain impingement. No differences were found concerning the initial posture of the thoracic spine. CONCLUSION: There is a relation between mobility of the thoracic spine and an impingement syndrome. This should be respected in diagnosis and therapy. PMID- 15346303 TI - [Cervical collars: a clinical and biomechanical study]. AB - AIM: Cervical collars have been used for years in the non-operative and postoperative management of cervical spine problems. They offer considerable differences in design, stability, bearing comfort and costs. In the background of controversies about the indication and in view of the limited number of scientific publications on the topic we have conceived this paper as a biomechanical and clinical investigation. METHODS: During the biomechanical investigations a defined load was exerted on 10 cervical collars of 4 producers. The clinical part was the measurement of the limitation of movement due to the 10 cervical collars in 30 healthy subjects aged 20 to 60 years. RESULTS: The biomechanical investigations showed differences of stiffness which were obvious during axial load, sideward and backward bending and less obvious in forward bending. The clinical study in 30 subjects indicated, on the other hand, only small differences in the limitation of movement. Softer cervical collars were slightly better accepted in the subjective judgment. CONCLUSION: For the aimed limitation of movement, a minimum of stiffness is obviously necessary, which is common with every tested collar. Higher degrees of stiffness do not result in further limitation of movement but only in a reduction of bearing comfort. PMID- 15346304 TI - [Mid-term clinical results of in-patient conservative treatment of cervicobrachialgia syndromes]. AB - AIM: The aim of this study was to evaluate the mid-term results of an in-patient conservative therapy regimen for discogenic cervicobrachialgia. METHODS: A retrospective follow-up of 100 patients after an average of 4.01 years with clinical examination and a questionnaire was performed. RESULTS: The subjective outcome rating for pain on a scale from 0 (pain-free) to 10 (maximum imaginable pain) was reduced from 7.91 at beginning of the therapy to 3.19 at the end of the therapy and to 1.5 at follow-up (p < 0.005). Duration of pain, reductions of motion and palsy improved significantly. The patients' appraisal of required surgical therapy could be reduced from 26 % at beginning to 2 % at the end of the therapy. CONCLUSION: The intensive conservative in-patient treatment for discogenic cervicobrachialgia shows a high satisfaction of the treated patients even after a follow-up of four years. The natural course of the disease shows a rapid and stable remission after wearing off the acute or subacute complaints. PMID- 15346305 TI - [Outcome sensitivity of the North American Spine Society Instrument with special consideration for the neurogenic symptoms of chronic back pain]. AB - BACKGROUND: Back pain is one of the most common chronic diseases in developed countries. The related enormous direct and indirect costs demand evidence-based decisions on rehabilitative care. Our study is intended to evaluate the two NASS outcome dimensions pain and neurological symptoms regarding their sensitivity to change within an inpatient rehabilitation scheme for chronic back pain. METHODS: The study observed a cohort of consecutive patients for one year (n = 70). By applying two outcomes instruments which are in common use internationally (SF-36 and NASS) it enhances comparability with other studies. Moreover the FFbH-R was applied for cross validation. RESULTS: On discharge from hospital an improvement of physical and emotional health as well as pain reduction were found. The results of the various instruments are consistent and support each other and suggest sensitivity to change of the NASS instrument regarding its pain and neurology subdimensions. CONCLUSION: The NASS outcome instrument monitors well the established therapeutic effects of chronic back pain inpatient rehabilitation schemes in the short run and in the long run over a twelve-month period following hospital discharge. PMID- 15346306 TI - [Outcome between surgical and non-surgical treatment of metastatic tumors of the spine: a retrospective study of 259 patients]. AB - AIM: The purpose of this study was to evaluate the clinical outcome of patients with metastatic tumors of the spine after surgical and non-surgical treatment. METHODS: The charts of 259 patients with metastatic tumors of the spine were reviewed retrospectively to define predictors of outcome. Our data included patient demographics, primary tumor, location of the metastatic tumor within the spine, indication for surgical or non-surgical treatment, type of surgical and non-surgical intervention, post-treatment outcome in terms of neurology, use of adjuvant radiation therapy or systemic therapy. RESULTS: The most frequent indication for surgical treatment was the combination of neurological deficit (ND), pathological vertebral fracture, and pain (50 %). Surgical intervention was performed by the posterior approach in 67 %, by the anterior approach in 13 %, and by an anterior/posterior approach in 10 %. The post-surgical outcome, depending on the type of surgical approach in terms of ND, was for the posterior approach 29 % improved, for the anterior approach 53 % improved, and for the anterior/posterior approach 15 % improved. DISCUSSION: Our data suggest that the indications for metastatic tumor surgery in the spine depend on the location of the metastatic tumor in the spine, clinical symptoms, and prognosis. PMID- 15346307 TI - [Vertebral body replacement with expandable titanium cages]. AB - OBJECTIVES: The aim of vertebral body replacement is the stabilisation and restoration of the anterior column of the spine with removal of the diseased region. We present our results of stabilisation, pain reduction and neurological improvement using vertebral-body replacement systems METHODS: Between April 1997 and December 2002, 53 patients with malignant vertebral destruction or instability due to traumatic and osteoporotic fracture were treated. We evaluated the results after vertebrectomy and vertebral body replacement by using expandable titanium cages in a retrospective study. RESULTS: The average follow up time was 18.9 +/- 19.9 months. The mean operation time was 173.2 +/- 77.4 minutes. Intraoperatively, we saw no implant-related complications. Perioperatively, complications appeared in 18 patients (34.0 %). 4 of them were severe, with 2 patients dying. In the total follow-up, 16 patients died, 10 of them (62.5 %) due to tumour progression. Pain reduction was reported in 52.7 %, neurological improvement in 48.0 %. One patient had a loosening of his posterior instrumentation in the further follow-up due to tumour growth with dislocation of the cage and a deterioration of his neurological deficit. CONCLUSION: By using vertebral body replacement systems, sufficient stabilisation of the vertebral column, pain reduction and neurological improvement can be achieved with an acceptable perioperative risk. PMID- 15346308 TI - [Minimally invasive therapy for osteoid osteoma]. AB - AIM: We describe a minimally invasive procedure for the treatment of osteoidosteoma and present the outcome of the treatment in our collective. Additionally, a review of the literature for minimally invasive therapy for osteoidosteoma was performed. METHODS: Within approximately three years, ten male patients with the clinical and radiological diagnosis of osteoidosteoma were treated. First, during the procedure for the purpose of orientation, a CT scan of the affected area was performed. Then, under CT-guidance, the nidus of the osteoidosteoma was marked with a k-wire and destroyed with a drill, moulding cutter or the k-wire. The patients' charts and films were reviewed in a retrospective manner, all patients participated in a personal telephone interview. RESULTS: In all cases the intraoperative course was free of complications. After surgery 90 % of the patients were free of pain, in 10 % a significant reduction of the pain was reported. Both in those 10 % and the whole collective there were no clinical symptoms of relapse, the average follow-up in all cases was 19 months. Our results are in accord with data in the literature. CONCLUSION: In our collective the CT-guided, percutaneous removal of osteoidosteoma has been shown to be a safe and effective method. Under DRG conditions it can be performed within a short period of hospitalization or in the out-patient clinic. PMID- 15346309 TI - [Antibacterial effectiveness of high-energetic extracorporeal shock waves: an in vitro verification]. AB - AIM: Extracorporeal shock wave therapy (ESWT) is used for a multitude of different indications in modern orthopedics. Local bacterial infections, like infected pseudarthrosis, are still considered as contraindications. The goal of the present study was to determine the effect of ESWT on the growth of clinically relevant bacteria in orthopedic and trauma surgery. METHODS: Standardized suspensions of five bacterial strains of bone and implant-associated infections were treated with 4 000 impulses of high-energy shock waves with an energy flux density (ED) of 0.96 mJ/mm (2) and a frequency of 2 Hz. Subsequently, viable bacteria were quantified and compared with an untreated control. RESULTS: A highly significant antibacterial effect of the ESWT was demonstrated for all bacterial strains with a reduction of growth to values between 1.1 % and 29.7 % (p < 0.01). Reference strains of Staphylococcus aureus and Staphylococcus epidermidis reacted with the highest sensitivity whereas Enterococcus faecium demonstrated the highest resistance towards high-energy shock waves. CONCLUSION: ESWT proved to exert a significant antibacterial effect on clinically relevant pathogens. Further investigations on energy flux density and impulse rates might contribute to an optimization of the bactericidal effectiveness. Infections as possible indications of the ESWT should therefore be assessed in further studies and the clinical relevance should be verified in an animal model. PMID- 15346310 TI - [Induction of bone tissue on different matrices: an in vitro and a in vivo pilot study in the SCID mouse]. AB - AIM: Three resorbable biomaterials were evaluated regarding proliferation and osteogenic differentiation of human bone marrow stromal cells (BMSC) in vitro. In a second step, the new biomaterial, calcium-deficient hydroxyapatite (CDHA), was tested in a pilot in vivo study by subcutaneous implantation in the severe combined immunodeficiency (SCID) mouse. METHODS: CDHA, beta-tricalcium phosphate (beta-TCP), and demineralized bone matrix (DBM) were seeded with human BMSC and cultured in osteogenic supplements for 3 weeks. In the pilot in vivo study, CDHA was seeded with BMSC and kept in osteogenic media for 2 weeks (group A) before subcutaneous implantation in 8 SCID mice for 3 and 8 weeks. In addition, CDHA seeded with BMSC without prior osteogenic induction (group B) and empty ceramics were implanted in each mouse. RESULTS: Total protein content and the values for specific alkaline phosphatase (ALP) increased significantly in vitro on all matrices, but no significant difference between the groups was noted. In the pilot in vivo study all ceramics were well penetrated by cells. After 8 weeks 2 of 4 samples in group B and 1 of 4 samples in group A revealed cells resembling hypertrophic chondrocytes. Specific ALP was higher in the group B (p = 0.012, Z = - 2.5) compared to empty ceramics. There were no significant differences between groups A and B. Differences between group A and the empty control did not become significant (p = 0.069, Z = - 1.8). CONCLUSION: All three matrices promoted BMSC proliferation and differentiation to osteogenic cells in vitro. Human BMSC on CDHA showed signs of osteogenic differentiation after subcutaneous implantation into SCID mice. PMID- 15346311 TI - [Normalization of histomorphometric and biomechanical parameters through metabolic control of fracture healing in spontaneously diabetic BB/OK rats]. AB - AIM: The objective of this study was to examine changes in the histomorphometrical and mechanical parameters of bone during fracture healing in dependence on the diabetic metabolic state in spontaneously diabetic BB/O(ttawa)K(arlsburg) rats. METHODS: Based on blood glucose values at the time of surgery, postoperative blood glucose course and postoperative insulin requirements, 90 spontaneously diabetic BB/OK rats were divided into groups with well compensated or poorly compensated metabolic states. Forty-five LEW.1A rats served as the normoglycemic controls. The femurs were fractured in a standardized procedure and then allowed to heal for 2, 4 and 6 weeks. RESULTS: In terms of bone histomorphometry, our study showed that, within the first four weeks after fracture, severe mineralization disorders occurred exclusively in the rats with poorly compensated diabetic metabolic states, with a significant decrease of all fluorochrome- based parameters of mineralization, apposition, formation and a significantly prolonged timing of mineralization, as well as significantly decreased values of biomechanical properties in comparison to the spontaneously diabetic rats with well compensated metabolic states and to the control rats. CONCLUSION: With a strictly controlled insulin therapy and a resulting well compensated diabetic metabolic state, severe mineralization disorders and the decreased values of biomechanical properties in the fracture repair in the poorly compensated diabetic metabolic state of the experimental animal can be avoided. PMID- 15346312 TI - [Secondary hypertrophic osteoarthropathy in colitis ulcerosa--a case demonstration]. AB - We report the case of a patient suffering from ulcerative colitis with secondary hypertrophic osteoarthropathy. We describe the pathological findings and present a model of the pathogenesis in the Strumpell-Pierre-Marie-Bamberger syndrome. PMID- 15346313 TI - [Intramuscular application of diclofenac--case report and critical consideration of a therapeutic measure]. AB - Diclofenac is a non-steroidal anti-inflammatory drug of the phenylacetic class. It is frequently given as an intramuscular injection. However several, sometimes severe, side effects have been described after an i.m. administration. Based on a short case report about a fatal complication in the context of the i.m. administration of diclofenac, the arguments for and against the intramuscular injection of the drug are critically discussed. As a result, the administration of diclofenac as an intramuscular injection should be critically reviewed and alternatives -- as suppositories are available -- should be taken into account. PMID- 15346314 TI - Subcranial transnasal repair of cerebrospinal fluid rhinorrhea with free autologous grafts by the combined overlay and underlay techniques. AB - OBJECTIVE: To determine the efficacy of subcranial transnasal repair of cerebrospinal fluid rhinorrhea with free autologous grafts by the combined overlay and underlay techniques using the surgical microscope and/or endoscope. PATIENTS AND METHODS: Twenty patients with CSF rhinorrhea were included in this retrospective study. They were 13 males and 7 females. Their age ranged from 7 to 62 years (mean: 39.35). The etiologies of the leak were iatrogenic in 10 cases, spontaneous in 5 cases, traumatic in 4 cases and one case was associated with meningeo-encephalocele. Preoperative nasal endoscopic examination, computed tomography (CT) with intrathecal non-ionic contrast and magnetic resonance imaging (MRI) were done when indicated. Endoscopic and/or microscopic repair of the CSF fistula was done by a combination of both underlay and overlay repair with free autologous grafts as follows: Gelfoam with fibrin glue, strips of fat, facia lata, Gelfoam with fibrin glue (underlay), septal cartilage, Gelfoam with fibrin glue and strips of fat (overlay). RESULTS: Complete closure of the leak was achieved in all patients. In one case of spontaneous CSF leak which was operated endoscopically, the leak recurred 6 months postoperatively and ceased spontaneously after a month with conservative medical treatment. No major complications were seen and no patients developed meningitis or postoperative anosmia. CONCLUSION: Subcranial transnasal repair with free autologous grafts by the combined overlay and underlay techniques using the endoscope or surgical microscope is a safe and successful method of treating CSF leaks, provided that the CSF leak is precisely located and the site can be reached with the endoscope or surgical microscope. PMID- 15346315 TI - Stereotactic radiosurgery without radiation therapy providing high local tumor control of multiple brain metastases from renal cell carcinoma. AB - The aim of this study was to analyse treatment effects after stereotactic radiosurgery (SRS) without whole brain radiation therapy (WBRT) as primary treatment for patients harboring brain metastases of renal cell carcinoma (RCC). During an 8-year period, 85 patients with 376 brain metastases from RCC underwent 134 outpatient SRS procedures. 65 % of all patients had multiple brain metastases. The median tumor volume was 1.2 cm (3) (range: 0.1 - 14.2 cm (3)). Mean prescribed tumor dose was 21.2 (+/- 3.2) Gy. Local/distant tumor recurrences were treated by additional SRS in cases of stable systemic disease. Overall median survival was 11.1 months after SRS. The local tumor control rate after SRS was 94 %. Most patients (78 %) died because of systemically progressing cancer. A KPS > 70 and RTOG class I were related to prolonged survival time. Patients of the RTOG groups I, II and III survived for 24.2 months, 9.2 months and 7.5 months, respectively. There was no permanent morbidity after SRS. 11 patients (12.9 %) showed transient radiogenic complications and 3 patients (3.5 %) died because of intratumoral bleedings after SRS. Stereotactic radiosurgery alone achieves excellent local tumor control rates for patients with small brain metastases from renal cell carcinoma. PMID- 15346316 TI - Endoscopic treatment of encephaloceles of the lateral wall of the sphenoid sinus. AB - OBJECTIVE: The suitability of the endoscopic approach for the treatment of an encephalocele of the lateral wall of the sphenoid is discussed. This is a retrospective review of 4 cases diagnosed with temporosphenoidal encephalocele and having a history of CSF leak who were surgically treated using an endoscopic endonasal approach between January 2001 and June 2002 at the Department of Otolaryngology of Sant'Orsola-Malpighi University Hospital and the Department of Neurosurgery at Bellaria Hospital in Bologna. METHODS: Three patients were female between 48 and 73 years of age (mean: 61 years). All patients had suffered from a CSF leak for 5 months to 18 years. None of the patients had a past medical history of head trauma. A fourth patient had undergone a previous microscopic approach for a previously misdiagnosed CSF leak wrongly ascribed to an empty sella. Three patients underwent an ethmoid-pterygo-sphenoidal endoscopic approach (EPSEA), while the patient who had undergone previous microscopic surgery, was treated using a transnasal transsphenoidal endoscopic approach. RESULTS: The follow-up of the patients ranged from 10 to 26 months (mean: 18 months) and no case of a recurrent CSF leak was observed postoperatively. CONCLUSIONS: In our report, the endoscopic approach was a useful tool for the treatment of encephaloceles of the lateral wall of the sphenoid sinus. In skilled hands, this technique permits both the resection of the encephalocele and the subsequent reconstruction of the defect also with a low rate of morbidity. PMID- 15346317 TI - Virtual endoscopy is a useful device for training and preoperative planning of transsphenoidal endoscopic pituitary surgery. AB - Virtual endoscopy (vE) allows simulated three-dimensional (3-D) visualisation of anatomical structures by computerised reconstruction of radiological images. The aim of this study was to evaluate the feasibility of vE and its potential benefits for endoscopic transsphenoidal pituitary surgery. vE was realised using a commercially available ray-casting software plugin of a picture archiving and communications system (PACS). For this study, the vE system was enhanced with volume segmentation, transparency and cutting tools. The data for vE were derived from high resolution computed tomography (CT) scans of 22 patients with pituitary pathology (20 pituitary adenomas, 2 Rathke's cleft cysts) preoperatively. Anatomic structures were identified on vE images and compared with the intraoperative endoscopic views. The simulated 3-D vE images were found to be comparable to the intraoperative endoscopic anatomy in terms of distortion and angle of view. vE was found to be particularly useful for the preoperative depiction of 1) the nasal anatomy and its variations for choosing the side of the approach, 2) the sphenoid sinus septae and chambers for improved intraoperative orientation, 3) the transparent 3-D simulated visualisation of the pituitary gland, tumour and adjacent anatomic structures in relation to the sphenoid sinus landmarks for planning the opening of the sellar floor. We conclude that vE harbours the potential to become a valuable tool in endoscopic pituitary surgery for training purposes and preoperative planning. Furthermore, vE may add to the safety of interventions in case of anatomic variations. PMID- 15346318 TI - The supraorbital keyhole approach via an eyebrow incision for resection of tumors around the sella and the anterior skull base. AB - OBJECTIVE: This study evaluates the technique, indications, advantages and limitations of the minimal invasive supraorbital keyhole approach via an eyebrow skin incision for resection of tumors around the sella and the anterior skull base. METHODS AND RESULTS: In 9 patients (5 males, 4 females) different tumors (6 meningiomas, 1 craniopharyngioma, 1 Rathke's cleft cyst and 1 hypophysitis) with a maximum diameter of 30 millimeters were resected via a small eyebrow incision and a supraorbital keyhole craniotomy. Details of the operative procedure include the use of a high-speed drill, a microsaw, bayonet-shaped instruments, careful microsurgical dissection, the use of a neuroendoscope and miniplates for closure. In all patients complete tumor removal was achieved. Mean duration of surgery was 01 h 51 min and mean hospital stay was 8.9 days. There were no significant postoperative complications. Clinical examination and MR imaging after a mean follow-up of 313 days revealed no tumor recurrence and a very satisfying overall functional and cosmetic result in all patients. CONCLUSION: The eyebrow incision supraorbital keyhole approach proved to be safe, effective and time-sparing. The authors recommend this approach for resection of small tumors around the sella and the anterior skull base in selected cases as a valuable alternative to standard skull base approaches. PMID- 15346319 TI - Spontaneous resolution of "protruded" lumbar discs. AB - Decreasing size of disc material that has herniated, whether "contained" or "protruded" has previously been described and sometimes this can be so complete that residual material is barely visible. In a retrospective clinical survey, from among almost 2180 consecutive patients admitted during 1994 - 2002 with low back pain; due to our low follow-up ratios and high price of the magnetic resonance imaging (MRI), only 42 patients with a lumbar disc protrusion could be found who had two MRI scans obtained at least five weeks apart. Among these, 4 patients were spotted with a totally resolved disc protrusion. T (2)-weighted MRI images were suggesting shrinkage due to dehydration and regression within the annulus of protruded disc fragments that had not fully migrated. Our patients are further examples for total resolution of the large "protruded" disc without any treatment; and since concomitant disc protrusions at other levels persisted, the resolution of these discs is supposed to be spontaneous. PMID- 15346320 TI - Endoscopic endonasal transsphenoidal removal of an intra-suprasellar schwannoma mimicking a pituitary adenoma. AB - Intracranial schwannomas, accounting for 8 to 10 % of all primary brain neoplasms, are relatively frequent intracranial tumors, but a "pure" intrasellar localization is exceptional. We report the case of an intra-suprasellar schwannoma mimicking a non-functioning pituitary macroadenoma both radiographically and clinically. A 73-year-old man presented with an episode of lipothymia followed by episodes of mental confusion. The neurological investigations revealed a bitemporal hemianopia and a hypopituitaric status. The neuroradiological investigations showed an intra-suprasellar mass resembling a pituitary adenoma. The patient underwent surgery performed by means of an endoscopic endonasal transsphenoidal approach, with a subtotal excision of the tumor. The histopathological studies revealed a cellular schwannoma. The review of the literature disclosed another 8 cases of intrasellar schwannomas. The possibility of an intrasellar schwannoma has to be considered in the differential diagnoses of neoplastic and non-neoplastic lesions of the sellar area. PMID- 15346321 TI - Relief of hemifacial spasm after radiosurgery for intracanalicular vestibular schwannoma. AB - Secondary hemifacial spasm due to vestibular schwannoma is very rare. This is the first reported case of hemifacial spasm responsive to gamma knife radiosurgery in a patient with an intracanalicular vestibular schwannoma. Both the resolution of the spasm as well as tumor growth control were achieved with a single session of gamma knife radiosurgery. We report a 49-year-old male patient with a 6-month history of right-sided hearing loss and hemifacial spasm. MR examination revealed an intracanalicular vestibular schwannoma. The patient was treated with radiosurgery and received 13 Gy to the 50 % isodose line. Tumor growth control was achieved and no change in the tumor volume was present at the last follow-up at 22 months. The hemifacial spasm completely resolved after one year. Surgical removal of the presumably causative mass lesion has been reported to be the sole treatment in secondary hemifacial spasm. This case report indicates that it may be responsive to gamma knife radiosurgery. Whether or not this might be a treatment option in selected refractory cases of hemifacial spasm remains to be defined. PMID- 15346322 TI - Minimally invasive management of the third ventricle glioma in a patient without hydrocephalus: neurofiberscopic biopsy followed by gamma knife radiosurgery. AB - There is usually concern that ventricular enlargement is necessary for safe neuroendoscopic procedures. However, newly developed models of neurofiberscopes with a small outer diameter can be effectively used even in patients without ventriculomegaly. We present the case of a successful neurofiberscopic biopsy of a third ventricle anaplastic astrocytoma in a previously shunted patient without hydrocephalus. Subsequent gamma knife radiosurgery led to complete disappearance of the tumor. It seems that presence of collapsed ventricles, and even symptoms consistent with the slit ventricle syndrome, should not be considered as contraindications for neurofiberscopic surgery when brain compliance is sufficiently preserved. The latter should be considered as the key point of safe endoscopic manipulations. The combination of the neuroendoscopy and stereotactic radiosurgery provides wide possibilities for minimally invasive management of selected intracranial tumors. PMID- 15346323 TI - The surgical management of traumatic C6 - C7 spondyloptosis in a patient without neurological deficits. AB - An unusual case of traumatic C6 - 7 total spondyloptosis with neurologically intactness at the time of injury is reported in a 35-year-old man. The patient was treated with a single-stage combined anterior-posterior and anterior operation to restore the cervical spondyloptosis, and creation of a three-column stabilization of the spine without neurological deficits. To the best of the authors' knowledge, there is no case report of traumatic spondyloptosis of cervical spine, presenting without neurological deficits in the pre- and postoperative periods. A brief summary of the clinical presentation, the surgical technique, and a review of the relevant literature are presented. PMID- 15346324 TI - Minicraniotomy treatment of an intracerebral epidermoid cyst. AB - Intracerebral epidermoid cysts are extremely rare. We report the case of a 16 year-old girl with an intracerebral epidermoid cyst in the left temporal lobe. The lesion was totally removed through a minimally invasive minicraniotomy by which we could obtain satisfactory intraoperative disclosure and manipulation. The incidence, clinical course, radiological features, surgical treatment, and complications of such a rare lesion are discussed. PMID- 15346325 TI - Combined three-dimensional anisotropy contrast imaging and magnetoencephalography guidance to preserve visual function in a patient with an occipital lobe tumor. AB - OBJECTIVE: Three-dimensional anisotropy contrast (3-DAC) magnetic resonance imaging and magnetoencephalography (MEG) of visually evoked magnetic fields (VEFs) were used to accurately localize the optic radiation and primary visual cortex before surgery for an occipital tumor. PATIENT AND METHODS: A 26-year-old male presented with an occipital lobe tumor located intrinsically underneath the right calcarine fissure. 3-DAC imaging showed that the right optic radiation was located along the superior and lateral surfaces of the lesion. Mapping of the VEFs demonstrated that the primary visual cortex was located superior and lateral to the lesion. The lesion was totally resected via an infero-medial cortical incision using a frameless stereotactic system. Histopathology indicated a pilocytic astrocytoma. No visual deficit was found before or after surgery. CONCLUSION: Combined 3-DAC imaging and MEG can provide essential information about the optic radiation and primary visual cortex for planning the surgical treatment of occipital lobe tumors. PMID- 15346326 TI - A case of severe low back pain associated with Richard disease (lumbosacral transitional vertebra). AB - The imaging findings in some cases of low back pain are difficult to describe and the pathophysiology is not clear because subjective findings are usually difficult to document. This report concerns a rare case about the lumbosacral joint causing persistent severe back pain which in itself does not usually have any pathological meaning. We describe the diagnosis, treatment, and outcome of a patient suffering from low back pain with lumbosacral transitional vertebra. The radiological examination showed the congenitally malformed transverse processes of the 5th lumbar vertebra to create a facet joint-like shape at the sacral ala region on both sides. Surgery resulted in immediate disappearance of the low back pain. When the lumbosacral junction shows some instability without fusion, the joint-like region could be the cause of low back pain. The lumbosacral transitional vertebra may be associated with low back pain. Electric denervation of the space between the transverse process of 5th lumbar vertebra and sacral ala is minimally invasive and effective for pain eradication. PMID- 15346327 TI - Non-traumatic compression fractures of the thoracic spine following a seizure -- treatment by percutaneous kyphoplasty. AB - A 26-year-old male presented with acute mid-thoracic back pain following a witnessed grand mal seizure. There was no trauma and the patient was on steroids for systemic lupus erythematosus. X-rays and CT scans of the thoracic spine revealed compression fractures at T5 and T6, with 50 % loss of vertebral height and kyphosis. He underwent percutaneous kyphoplasty of both vertebrae, with symptomatic improvement. Non-traumatic compression fractures of the thoracic spine following seizures are a rare injury. This may be related to the compressive forces exerted on the vertebral column by the contractions of the truncal muscles, during a seizure. These compression fractures are suitable for treatment by minimally invasive techniques, such as kyphoplasty. PMID- 15346328 TI - Thecaloscopy/Thecoscopy. PMID- 15346329 TI - Temporal and geographic stability of the serogroup-specific invasive disease potential of Streptococcus pneumoniae in children. AB - A meta-analysis study design was used to analyze 7 data sets of invasive and carriage pneumococcal isolates recovered from children, to determine whether invasive disease potential differs for each serotype and, if so, whether it has changed over time or differs geographically. Serotype- and serogroup-specific odds ratios (ORs) were calculated for each study and as a pooled estimate, with use of serotype 14 as the reference group. ORs varied widely: the serotypes with the highest ORs (1, 5, and 7) were 60-fold more invasive than those with the lowest ORs (3, 6A, and 15). There was a significant inverse correlation between invasive disease and carriage prevalence for the serotypes that we considered, which implies that the most invasive serotypes and serogroups were the least commonly carried and that the most frequently carried were the least likely to cause invasive disease. There was no evidence of any temporal change or major geographical differences in serotype- or serogroup-specific invasive disease potential. PMID- 15346330 TI - Differences in survival, brain damage, and cerebrospinal fluid cytokine kinetics due to meningitis caused by 3 different Streptococcus pneumoniae serotypes: evaluation in humans and in 2 experimental models. AB - BACKGROUND: Experimental meningitis with Streptococcus pneumoniae serotypes 1, 3, and 9 has resulted in pronounced differences in disease severity, but clinical meningitis studies addressing serotype-related differences in case-fatality rates are lacking. METHODS: Study subjects were Danish patients with pneumococcal meningitis due to serotype 1 (n=38), 3 (n=69), or 9V (n=59) during 1990-2002 for whom clinical information was available. The 3 serotypes were tested for brain damage and cerebrospinal fluid (CSF) inflammatory kinetics in 2 experimental models of meningitis. RESULTS: Patients with serotype 1 had a significantly lower case-fatality rate (3%), compared with patients with serotypes 3 (23%) and 9V (32%) (P=.0047, log-rank test). Age and serotype were independent prognostic factors for fatal outcome. In experimental meningitis, the median number of areas per brain slide with brain damage was significantly lower in rats infected with serotype 1 than in rats infected with serotypes 3 and 9V. Three distinct patterns of brain damage were observed: serotype 1, cortical hemorrhage; serotype 3, cortical necrosis and abscess formation; and serotype 9V, subcortical (callosal) abscess formation. Serotype 1 caused the poorest bacterial growth and lowest CSF levels of white blood cells, tumor necrosis factor- alpha, and interleukin-8 (P<.05). CONCLUSION: Case-fatality rates of patients with pneumococcal meningitis, the degree and pattern of brain damage, and CSF cytochemical alterations in experimental pneumococcal meningitis differ according to serotype. PMID- 15346331 TI - T cells mediate cross-protective immunity between spotted fever group rickettsiae and typhus group rickettsiae. AB - Rickettsioses are severe infections caused by obligately intracellular bacteria that preferentially infect the endothelium lining the vasculature. The causative agents, rickettsiae, have been divided according to biological, genetic, and antigenic parameters into 2 main groups: spotted fever and typhus. They have not been thought to stimulate cross-reactive protective immune responses; however, in this study, we show that, in relevant animal models that mimic human rickettsial infections, there is reciprocal immunological cross-protection between spotted fever group and typhus group rickettsiae. Furthermore, we present evidence that T cells are responsible for this cross-immunity and that cross-stimulation of T cells also occurs in humans. PMID- 15346332 TI - Immune responses to Bacillus anthracis protective antigen in patients with bioterrorism-related cutaneous or inhalation anthrax. AB - Anti-protective antigen (PA) immunoglobulin (Ig) G, toxin neutralization, and PA specific IgG memory B cell responses were studied in patients with bioterrorism related cutaneous or inhalation anthrax and in a patient with laboratory-acquired cutaneous anthrax. Responses were determined for >1 year after the onset of symptoms. Eleven days after the onset of symptoms (15 days after likely exposure), anti-PA IgG was detected in 16 of 17 patients with confirmed or suspected clinical anthrax who were tested. Anti-PA IgG remained detectable 8-16 months after the onset of symptoms in all 6 survivors of inhalation anthrax and in 7 of 11 survivors of cutaneous anthrax who were tested. Anti-PA IgG levels and serum toxin neutralizing activity were strongly associated (R2=0.83). PA-specific IgG memory B cells were detectable in all 6 survivors of inhalation anthrax but in only 2 of 7 patients with cutaneous anthrax who were tested. Anti-PA IgG is an important diagnostic marker of anthrax, a predictor of serum anti-toxin activity, and a marker of immunological memory against anthrax. PMID- 15346333 TI - Regions of high antigenicity within the hypothetical PPE major polymorphic tandem repeat open-reading frame, Rv2608, show a differential humoral response and a low T cell response in various categories of patients with tuberculosis. AB - The function of the PE/PPE families of proteins, which represent approximately 10% of the coding capacity of the Mycobacterium tuberculosis genome, has remained relatively unknown. We earlier described a PPE family member, Rv2430c, as an immunodominant antigen. We now report another PPE family gene, Rv2608, a member of the major polymorphic tandem repeat subfamily, for its ability to elicit a high humoral and a low T cell response. Rv2608 was also found to be polymorphic in different clinical isolates of M. tuberculosis, as determined by polymerase chain reaction-restriction fragment-length polymorphism analysis. A total of 51 clinically confirmed patients with tuberculosis (TB), belonging to 3 different categories--fresh infection (n=22), relapsed infection (n=21), and extrapulmonary infection (n=8)--and 10 healthy control subjects were included in the study. Recombinant Rv2608 protein showed positive reactivity to patients' serum samples. Enzyme-linked immunosorbent assays and T cell-proliferation assays with synthetic peptides corresponding to predicted regions of high antigenicity showed a predominantly humoral response in patients with relapsed TB. We additionally identified the Gly-X-Gly-Asn-X-Gly repeat motifs as being primarily responsible for eliciting a humoral immune response. PMID- 15346334 TI - Loss of microbicidal activity and increased formation of biofilm due to decreased lactoferrin activity in patients with cystic fibrosis. AB - Intractable formation of biofilm by and infection with the opportunistic pathogen Pseudomonas aeruginosa are hallmarks of cystic fibrosis (CF). Lactoferrin, an innate immunity protein, has recently been shown to inhibit the formation of P. aeruginosa biofilm. Partial cleavage of lactoferrin by the proteases neutrophil elastase and Pseudomonas elastase has previously been described in CF. Here, we show that cathepsins in CF secretions are responsible for complete and rapid cleavage of lactoferrin. We demonstrate that levels of lactoferrin in P. aeruginosa-positive sputum samples are decreased when corrected for inflammatory burden and that P. aeruginosa-positive sputum samples have significantly higher cathepsin activity and significantly reduced ability to inhibit formation of biofilm, compared with P. aeruginosa-negative sputum samples. We also show that cleavage of lactoferrin by cathepsin results in loss of both its microbicidal and antibiofilm activity. Loss of such a vital innate immunity protein clearly has important implications for the pathogenesis of chronic P. aeruginosa lung infection in patients with CF. PMID- 15346335 TI - Cross-reactivity of human immunoglobulin G2 recognizing phosphorylcholine and evidence for protection against major bacterial pathogens of the human respiratory tract. AB - Phosphorylcholine (ChoP) is an antigenic component on the cell surface of many commensal and pathogenic bacteria that reside in the upper airway. In the present study, human ChoP-specific antibody was affinity-purified from pooled serum gamma globulin. This naturally acquired antibody, which is primarily of the immunoglobulin (Ig) G2 subtype, recognized ChoP on the lipoteichoic acid of Streptococcus pneumoniae and on the lipopolysaccharide of Haemophilus influenzae, 2 of the leading etiologic agents of infection involving the human respiratory tract. In in vitro killing assays, anti-ChoP IgG2 was effective against some clinical isolates of nontypeable H. influenzae and against isolates of several common serotypes of S. pneumoniae. Moreover, passively administered human anti ChoP antibody protected mice against lethal challenge with a transparent isolate of S. pneumoniae type 6A. The effectiveness of human antibody to this conserved bacterial structure suggests that, if it can be manipulated to broaden its activity, it could function as a single vaccine antigen that targets multiple pathogens. PMID- 15346336 TI - Long-term persistence of immunity to hepatitis B after vaccination during infancy in a country where endemicity is low. AB - BACKGROUND: The long-term response to hepatitis B vaccination during infancy has not been fully evaluated in countries where endemicity is low. METHODS: The present study was a serological investigation of immunity to hepatitis B during adolescence. In a cohort of children who were born to hepatitis B virus carrier mothers and who were vaccinated during infancy, evidence of past or current infection and the response to a single booster dose of vaccine were analyzed. Sixty-four children whose antibody levels were measured after immunization (group 1) and 52 younger siblings who did not undergo postvaccination antibody tests (group 2) were studied. RESULTS: One child in each group showed evidence of natural infection. In group 1, 32 children (50%) had undetectable antibody to hepatitis B surface antigen (anti-HBs), and only 8 (13%) had levels >100 mIU/mL. After a booster dose of vaccine, only 7 (11%) still had undetectable anti-HBs, 3 (5%) showed a primary response, and 50 (78%) had levels >100 mIU/mL. Five of the 7 vaccine nonresponders and all of the primary responders had also received hepatitis B-specific immunoglobulin (HBIG) at birth. The children in group 2 showed a similar response to the vaccine, but with slightly higher levels of anti HBs. CONCLUSIONS: Most children vaccinated at birth retain immunological memory to hepatitis B vaccine for 15 years, but those who did not were more likely to have received HBIG at birth, suggesting that passive antibody may interfere with the induction of immunological memory. PMID- 15346337 TI - Clearance of hepatitis C virus after newly acquired infection in injection drug users. AB - A retrospective cohort of injection drug users with newly acquired hepatitis C virus (HCV) infection was established to examine viral clearance. Newly acquired HCV infection was defined by anti-HCV antibody seroconversion within a 2-year interval. Stored serum samples were tested for HCV RNA, with viral clearance defined as >/=2 consecutive negative HCV RNA test results after infection. Ninety nine cases of HCV infection were identified; 57 had >/=2 HCV RNA test results after infection. Viral clearance occurred in 24 (42%) cases, with Kaplan-Meier estimated probabilities of 23%, 38%, and 40% at 6, 12, and 24 months, respectively. PMID- 15346338 TI - Provirus load in breast milk and risk of mother-to-child transmission of human T lymphotropic virus type I. AB - In a prospective study of 101 mother-child pairs in Jamaica, we examined the association of provirus load in breast milk and the risk of mother-to-child transmission of human T lymphotropic virus type I. The provirus load in breast milk was a strong predictor of risk of transmission to children (relative risk, 2.34/quartile), after adjustment for other known risk factors. The risk of transmission increased from 4.7/1000 person-months when the provirus load in breast milk was <0.18% to 28.7/1000 person-months when it was >1.5%. Provirus detection in maternal breast milk predicted transmission months before infection in children was detected by serologic testing. PMID- 15346339 TI - Polymorphism in the interleukin-10 promoter affects both provirus load and the risk of human T lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis. AB - To investigate non-human leukocyte antigen candidate genes that influence the outcome of human T cell lymphotropic virus (HTLV) type I infection, we analyzed 6 single-nucleotide polymorphisms in the interleukin (IL)-10 promoter region in 280 patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and 255 HTLV-I-seropositive asymptomatic carriers from an area where HTLV-I is endemic. The IL-10 -592 A allele, which shows lower HTLV-I Tax-induced transcriptional activity than the C allele in the Jurkat T cell line, was associated with a >2-fold reduction in the odds of developing HAM/TSP (P=.011; odds ratio [OR], 0.50 [95% confidence interval, 0.30-0.86]) by reducing the provirus load in the whole cohort (P=.009, analysis of variance). Given the OR and the observed frequency of IL-10 -592 A, we demonstrate that this allele prevents approximately 44.7% (standard deviation, +/-13.1%) of potential cases of HAM/TSP, which indicates that it defines one component of the genetic susceptibility to HAM/TSP in the cohort. PMID- 15346340 TI - Induction of human T cell-mediated immune responses after primary and secondary smallpox vaccination. AB - BACKGROUND: Postexposure vaccination strategies rely on a rapid induction of poxvirus-specific immune responses. Postvaccination cell-mediated immune (CMI) responses have not been compared by use of controlled trials in previously vaccinated (vaccinia-nonnaive) and nonvaccinated (vaccinia-naive) individuals. METHODS: To assess the time course of vaccinia-specific CMI responses, 20 previously vaccinated and 10 vaccinia-naive individuals were vaccinated with Dryvax, and serial blood samples were drawn. RESULTS: Both groups developed peak levels of vaccinia-specific interferon (IFN)- gamma -producing T cells by day 14 after vaccination. In vaccinia-nonnaive individuals, vaccinia-specific CMI responses were detected by day 7 after vaccination and preceded the increase in antibody titers. IFN- gamma enzyme-linked immunospot responses were significantly different between the 2 groups on days 7 (greater in vaccinia-nonnaive than in vaccinia-naive individuals) and 14 (greater in vaccinia-naive than in vaccinia nonnaive individuals). Lymphoproliferation responses in vaccinia-nonnaive individuals were significantly higher on days 3 and 7, but cytotoxic T cell lysis activity was not statistically different at any time point. Antibody responses conformed to expected primary and secondary patterns of induction. CONCLUSIONS: This study demonstrates that the kinetics of CMI responses are different after primary vaccination versus after revaccination and indicates that memory can exist in individuals vaccinated >/=30 years ago. These data support the epidemiological observation in smallpox outbreaks that successful revaccination within 4 days of exposure is partially protective. In vaccinia-nonnaive individuals, protection against smallpox during the postexposure revaccination period may require T cell memory as an essential component for the rapid induction of protective cellular and humoral responses. PMID- 15346342 TI - Codetection of a mixed population of candHPV62 containing wild-type and disrupted E1 open-reading frame in a 45-year-old woman with normal cytology. AB - We have cloned, sequenced, and characterized the complete genome of a novel human papillomavirus (HPV), candHPV62. During cloning, 2 candHPV62 viral isolates were recovered from a single cervical sample; 1 had all anticipated HPV open-reading frames (ORFs) intact, whereas the other exhibited an E1 frame-shift mutation. Further experiments indicated that the 2 strains were equivalent in abundance. It appears that an early mutation occurred within the E1 ORF, which was transcomplemented by an intact E1 protein. A search of the HPV database identified disruption of the E1 ORF in the cloned reference isolates of HPV16, HPV53, HPV56, and HPV72. These data suggest that disruption of the E1 ORF in genital HPVs is not uncommon. PMID- 15346341 TI - Clinical and immune responses after revaccination of israeli adults with the Lister strain of vaccinia virus. AB - BACKGROUND: During the winter of 2002-2003, the Israeli health authorities launched a campaign to vaccinate first responders against smallpox. METHODS: In an open study, 159 healthy, preimmunized adults, 24-52 years old, who participated in the campaign were vaccinated with the Lister strain of vaccinia virus by the multipuncture technique. The safety, immunogenicity, and reactogenicity of the vaccine were assessed. RESULTS: Successful vaccination rates were 61% and 56%, on the basis of clinical take and seroconversion, respectively. Adverse events among the vaccinees were minor. Seventy-nine (88%) of the 90 vaccinees with clinical take also seroconverted ( kappa =0.779). The level of preexisting antibodies inversely correlated with the rates of clinical take and seroconversion (P or = 20 mm Hg and/or decrease in diastolic BP > or = 10 mm Hg) was reported in another 13% patients. Perindopril/indapamid treatment led to a significant decrease in interventricular thickness (10.7 vs. 10.2, P < 0.01) and to the improvement in the left ventricle diastolic function (E/A 1.06 vs. 1,14, P < 0.04). No significant changes in basal laboratory tests were reported during the study. Mild side effects were observed in a few patients. CONCLUSION: Fixed, low-dose perindopril/indapamide combination is a new and potent antihypertensive compound with very good tolerability without a negative influence on metabolic parameters. Perindopril/indapamide treatment resulted in the improvement of diastolic function and in the regression of the left ventricle mass. PMID- 15346639 TI - [Infection and hemostasis]. AB - Localised and following systemic inflammatory reaction accompanying progression of infection causes generation of anti-inflammatory cytokines. They activate leucocytes, endothelium, coagulation and fibrinolysis. Sepsis is usually accompanied by already decompensated disseminated intravascular coagulation which significantly affects mortality of patients with this disease. The main cause of hypercoagulation state during sepsis seems to be inhibition of fibrinolysis as a result of overproduction of plasminogen activator inhibitor-1 in later stages of the disease. Some microorganisms have specific properties which affect individual components of hemostasis and thus increase their virulence. Because natural inhibitors of coagulation have not only anticoagulation but also strong anti inflammatory effect, they seem to be an optimum remedy for fluorid coagulopathy during sepsis. Moreover, their use usually does not increase risk of bleeding. PMID- 15346640 TI - [Clinical importance of aspirin resistance]. AB - The antiplatelet effect of aspirin is mostly explained by the irreversible cyclooxygenase-1 inhibition resulting in the suppression of thromboxane A2 synthesis. The benefit of aspirin was proved in various cardiovascular diseases. However, the inter- and intraindividual variability of its antiplatelet effect is well known. Aspirin resistance can be understood from the clinical point of view- as a failure of the protective effect of aspirin from thrombotic complication or can be defined from the laboratory aspect--as an inability to cause in vitro detectable platelet function inhibition. The cause of this phenomenon has not been completely explained yet and more mechanisms have been proposed, incomplete suppression of thromboxane A2 generation being one of them. Laboratory diagnostics of aspirin resistance is based on the demonstration of the insufficient inhibition of platelet aggregation or the incomplete suppression of thromboxane A2 synthesis (assay for its metabolite, 11-dehydrothromboxane B2 in urine). The results of some trials raise the possibility that aspirin resistance could be a new independent predictor of cardiovascular events. PMID- 15346641 TI - [Families at risk of colon cancer I. Familial adenomatous polyposis]. AB - Familial adenomatous polyposis (FAP) is a well-defined autosomal dominant inherited disease characterised by a diffuse polyposis of the colon and rectum leading to inevitable colorectal cancer by 50 years. The purpose of this review is to summarize the current knowledge regarding this entity with the focus on recent knowledge on genetic testing, surveillance guidelines and therapy of FAP. Available medical databases were searched from 1998 to May 2003 using keywords "familial adenomatous polyposis", followed by further search for particular issues. Additional articles were identified through the reference sections of retrieved articles and from personal archives of authors. Approximately 300 papers on FAP are published yearly. There has been a large progress in our understanding of the genetics of FAP leading to the development of genetic counselling, reliable genetic tests and screening strategies. There is accumulating evidence about genotype-phenotype associations with direct clinical implications. Our knowledge about the extracolonic manifestations is also expanding resulting in new surveillance and treatment strategies for FAP patients after proctocolectomy. Although still representing a serious burden for affected patients and their families, the research of last decades together with national registers improved the life expectancy and the quality of life of FAP patients dramatically. Further research in the area of molecular genetics, genetic testing and emerging gene therapy for FAP patients is to be expected in the near future. PMID- 15346642 TI - [Comment on the "European Stroke Initiative" (EUSI) and "Recommendations for care of patients with cerebrovascular stroke", version 2002]. PMID- 15346643 TI - [Comment by an arrhythmia specialist on the "European Stroke Initiative" (EUSI) and Recommendations on the care of patients with cerebrovascular stroke" version 2002]. PMID- 15346644 TI - Is serum gamma glutamyltransferase a marker of oxidative stress? AB - The primary role of cellular gamma glutamyltransferase (GGT) is to metabolize extracellular reduced glutathione (GSH), allowing for precursor amino acids to be assimilated and reutilized for intracellular GSH synthesis. Paradoxically, recent experimental studies indicate that cellular GGT may also be involved in the generation of reactive oxygen species in the presence of iron or other transition metals. Although the relationship between cellular GGT and serum GGT is not known and serum GGT activity has been commonly used as a marker for excessive alcohol consumption or liver diseases, our series of epidemiological studies consistently suggest that serum GGT within its normal range might be an early and sensitive enzyme related to oxidative stress. For example, serum and dietary antioxidant vitamins had inverse, dose-response relations to serum GGT level within its normal range, whereas dietary heme iron was positively related to serum GGT level. More importantly, serum GGT level within its normal range positively predicted F2-isoprostanes, an oxidative damage product of arachidonic acid, and fibrinogen and C-reactive protein, markers of inflammation, which were measured 5 or 15 years later, in dose-response manners. These findings suggest that strong associations of serum GGT with many cardiovascular risk factors and/or events might be explained by a mechanism related to oxidative stress. Even though studies on serum and/or cellular GGT is at a beginning stage, our epidemiological findings suggest that serum GGT might be useful in studying oxidative stress related issues in both epidemiological and clinical settings. PMID- 15346645 TI - Dual role of oxidized LDL on the NF-kappaB signaling pathway. AB - Atherosclerosis is a slowly evolutive age-linked disease of large arteries, characterized by a local lipid deposition associated with a chronic inflammatory response, leading potentially to acute plaque rupture, thrombosis and ischemic heart disease. Atherogenesis is a complex sequence of events associating first expression of adhesion molecules, recruitment of mononuclear cells to the endothelium, local activation of leukocytes and inflammation, lipid accumulation and foam cell formation. Low density lipoproteins (LDLs) become atherogenic after undergoing oxidation by vascular cells, that transform them into highly bioreactive oxidized LDL (oxidized LDLs). Oxidized LDLs are involved in foam cell formation, and trigger proatherogenic events such as overexpression of adhesion molecules, chemoattractant agents growth factors and cytokines involved in the inflammatory process, cell proliferation and apoptosis. Moreover, this toxic effect of oxidized LDLs plays probably a role in plaque erosion/rupture and subsequent atherothrombosis. Several biological effects of oxidized LDLs are mediated through changes in the activity of transcription factors and subsequently in gene expression. Oxidized LDLs exert a biphasic effect on the redox-sensitive transcription factor NF-kappaB, which can be activated thereby up regulating proinflammatory gene expression, such as adhesion molecules, tissue factor, scavenger receptor LOX-1. On the other hand, higher concentrations of oxidized LDLs may inhibit NF-kappaB activation triggered by inflammatory agents such as LPS, and may thereby exert an immunosuppressive effect. This review is an attempt to clarify the mechanism by which oxidized LDLs may up- or down-regulate NF-kappaB, the role of NF-kappaB activation (or inhibition), and the consequences of the oxidized LDLs-mediated NF-kappaB dysregulation and their potential involvement in atherosclerosis. PMID- 15346646 TI - Paraoxonase genotype, LDL-oxidation and carotid atherosclerosis in male life-long smokers. AB - Paraoxonase (PON-1) is a high-density lipoprotein (HDL) associated enzyme that hydrolyzes lipid peroxides in vitro, which may therefore protect against the onset of atherosclerosis. Heavy smokers are more exposed to oxidative stress and hence at high-risk for oxidative modification of LDL. Our hypothesis is that the anti-oxidative properties of PON-1 inhibit LDL oxidation, especially in populations exposed to high oxidative stress. We have studied the effects of PON 1 genotype and smoking to variation in oxidative status parameters and intima media thickness (IMT), a surrogate marker of atherosclerosis. The contribution of two common polymorphisms in the PON-1 gene (Q192R and L55M) to LDL oxidizability, autoantibodies directed against oxLDL and IMT were studied in 207 male life-long smokers. Smokers were classified into average, heavy and excessive smokers based on pack years of cigarettes smoked. PON-1 genotype was not associated with autoantibodies to oxLDL, LDL oxidizability or IMT. Smoking was associated with IMT in subgroups with the high levels of LDL, but not in the population at large. The lack of association of PON-1 genotype with oxidative status parameters and IMT suggests that PON-1 is not a major inhibitor of LDL oxidation in a population of life-long smokers. PMID- 15346647 TI - Investigation of the molecular nature of low-molecular-mass cobalt(II) ions in isolated osteoarthritic knee-joint synovial fluid. AB - High field 1H NMR spectroscopy demonstrated that addition of Co(II) ions to osteoarthritic knee-joint synovial fluid (SF) resulted in its complexation by a range of biomolecules, the relative efficacies of these complexants/chelators being citrate >> histidine - threonine >> glycine - glutamate - glutamine - phenylalanine tyrosine > formate > lactate >> alanine > valine > acetate > pyruvate > creatinine, this order reflecting the ability of these ligands to compete for the available Co(II) in terms of (1) thermodynamic equilibrium constants for the formation of their complexes and (2) their SF concentrations. Since many of these SF Co(II) complexants (e.g. histidinate) serve as powerful *OH scavengers, the results acquired indicate that any of this radical generated from the Co(II) source in such complexes via Fenton or pseudo-Fenton reaction systems will be "site-specifically" scavenged. The significance of these observations with regard to cobalt toxicity and the in vivo corrosion of cobalt containing metal alloy joint prostheses (e.g. CoCr alloys) is discussed. PMID- 15346648 TI - Oxidative status in chronic hepatitis C: the influence of antiviral therapy and prognostic value of serum hydroperoxide assay. AB - The effect of alpha-interferon (alpha-IFN) and ribavirin (RBV) treatment on oxidative status in chronic hepatitis C (CHC) is unknown. AIM: To study the time course of oxidative status in patients with CHC during alpha-IFN and RBV administration, and to evaluate the role of oxidative status in order to predict the therapeutic response. PATIENTS AND METHODS: Fifty one patients with CHC were studied. All received a combination of alpha-IFN and RBV for 6 or 12 months in relation to the type of response. The hydroperoxides concentration in serum test samples by D-ROM test was measured in all of the patients before therapy. In 27 patients, hydroperoxides were also measured during the treatment and during the 12 subsequent months. RESULTS: Cross-sectional analysis demonstrates that patients with a successive long-term response had a lower basal serum hydroperoxide concentration than non-responders (280 +/- 40.8 vs 337 +/- 83 CARR Units, p < 0.05). This resulted to be an independent factor predictive of long term response in the multi-varied analysis. Longitudinal observation on 27 patients showed that the mean hydroperoxide concentration decreased significantly during treatment (T0 329 +/- 79.2 vs T12 272 +/- 34.5 CARR Units) and that the decrease in the mean values was mainly due to variations in the relapsers group. CONCLUSIONS: Normal basal hydroperoxide concentration helps to predict long-term response to combination therapy. The D-ROM test may be used for screening patients before treatment. PMID- 15346649 TI - Nitrite generates an oxidant stress and increases nitric oxide in EA.hy926 endothelial cells. AB - Nitrite is a breakdown product of nitric oxide that in turn is oxidized to nitrate in cells. In this work, we investigated whether reactive oxidant species mightbe generated during nitrite metabolism in cultured EA.hy926 endothelial cells. Nitrite was taken up by the cells in a time- and concentration-dependent manner and oxidized to nitrate, which accumulated in cells to concentrations almost 10-fold those of nitrite. Conversion of low millimolar concentrations of nitrite to nitrate was associated with increased oxidant stress in the cells. This manifested as increased oxidation of dihydrofluorescein in tandem with depletion of both GSH and ascorbate. Further, loading cells with ascorbate or treatment with desferrioxamine prevented nitrite-induced dihydrofluorescein oxidation. Nitrite within cells also increased the fluorescence of 4-amino-5 methylamino-2',7'-difluorofluorescein and inhibited the activity of cellular glyceraldehyde 3-phosphate dehydrogenase, which are markers of intracellular nitrosation reactions. Intracellular ascorbate partially prevented both of these effects of nitrite. Although ascorbate can reduce nitrite to nitric oxide at low pH, in endothelial cells loaded with ascorbate, its predominant effect at high nitrite concentrations is to prevent potentially damaging nitrosation reactions. PMID- 15346650 TI - Do mitochondria make nitric oxide? no? AB - Several papers have claimed that mitochondria contain nitric oxide synthase (NOS) and make nitric oxide (NO*) in amounts sufficient to affect mitochondrial respiration. However, we found that the addition of L-arginine or the NOS inhibitor L-NMMA to intact rat liver mitochondria did not have any effect on the respiratory rate in both State 3 and State 4. We did not detect mitochondrial NO* production by the oxymyoglobin oxidation assay, or electrochemically using an NO* electrode. An apparent NO* production detected by the Griess assay was identified as an artifact. NO* generated by eNOS added to the mitochondria could easily be detected, although succinate-supplemented mitochondria appeared to consume NO*. Our data show that NO* production by normal rat liver mitochondria cannot be detected in our laboratory, even though the levels of production claimed in the literature should easily have been measured by the techniques used. The implications for the putative mitochondrial NOS are discussed. PMID- 15346651 TI - Reduction of DNA fragmentation and hydroxyl radical production by hyaluronic acid and chondroitin-4-sulphate in iron plus ascorbate-induced oxidative stress in fibroblast cultures. AB - Glycosaminoglycans (GAGs), components of extracellular matrix, are thought to play important roles in cell proliferation and differentiation in the repair process of injured tissue. Oxidative stress is one of the most frequent causes of tissue and cell injury and the consequent lipid peroxidation is the main manifestation of free radical damage. It has been found to play an important role in the evolution of cell death. Since several reports have shown that hyaluronic acid (HYA) and chondroitin-4-sulphate (C4S) are able to inhibit lipid peroxidation during oxidative stress, We investigated the antioxidant capacity of these GAGs in reducing oxidative damage in fibroblast cultures. Free radicals production was induced by the oxidizing system employing iron (Fe2+) plus ascorbate. We evaluated cell death, membrane lipid peroxidation, DNA damage, protein oxidation, hydroxyl radical (OH*) generation and endogenous antioxidant depletion in human skin fibroblast cultures. The exposition of fibroblasts to FeSO4 and ascorbate caused inhibition of cell growth and cell death, increased OH* production determined by the aromatic trap method; furthermore it caused DNA strand breaks and protein oxidation as shown by the DNA fragments analysis and protein carbonyl content, respectively. Moreover, it enhanced lipid peroxidation evaluated by the analysis of conjugated dienes (CD) and decreased antioxidant defenses assayed by means of measurement of superoxide dismutase (SOD) and catalase (CAT) activities. When fibroblasts were treated with two different doses of HYA or C4S a protective effect, following oxidative stress induction, was shown. In fact these GAGs were able to limit cell death, reduced DNA fragmentation and protein oxidation, decreased OH* generation, inhibited lipid peroxidation and improved antioxidant defenses. Our results confirm the antioxidant activity of HYA and C4S and this could represent a useful step in the understanding of the exact role played by GAGs in living organisms. PMID- 15346652 TI - Involvement of ceramide in the mechanism of Cr(VI)-induced apoptosis of CHO cells. AB - Mitochondria reduce Cr(VI) to Cr(V) with concomitant generation of reactive oxygen species, thereby exhibiting cytotoxic effects leading to apoptosis in various types of cells. To clarify the mechanism by which Cr(VI) induces apoptosis, we examined the effect of Cr(VI) on Chinese hamster ovary (CHO) cells. Cr(VI) increased cellular levels of ceramide by activating acid sphingomyelinase (ASMase) and inhibiting the phosphorylation of pleckstrin homology domain containing protein kinase B (Akt). Cr(VI) also induced cyclosporin A- and trifluoperazine-sensitive depolarization of mitochondria and activated caspase-3, 8 and 9, thereby causing fragmentation of cellular DNA. The presence of desipramine, an inhibitor of ASMase, and membrane permeable pCPT-cAMP suppressed the Cr(VI)-induced activation of caspases and DNA fragmentation. These results suggested that accumulation of ceramide play an important role in the Cr(VI) induced apoptosis of CHO cells through activation of mitochondrial membrane permeability transition. PMID- 15346653 TI - Oxidation of uric acid in rheumatoid arthritis: is allantoin a marker of oxidative stress? AB - Free radicals are implicated in many diseases including atherosclerosis, cancer and also in rheumatoid arthritis. Reaction of uric acid with free radicals, such as hydroxyl radical and hypochlorous acid (HOCl) results in allantoin production. In this study, we measured the serum allantoin levels, oxidation products of uric acid, as a marker of free radical generation in rheumatoid arthritis. Fasting blood samples were obtained from 21 rheumatoid patients and 15 healthy controls. In this study, the serum allantoin and uric acid levels were measured by a gas chromatography-mass spectrometry method and the ratios were calculated. The mean allantoin and uric acid levels and ratios in the patient group were 22.1 +/- 11.3, 280.5 +/- 65.0 and 8.0 +/- 3.7 microM, while in the control group they were 13.6 +/- 6.3, 278.3 +/- 53.6 and 4.9 +/- 2.1 microM, respectively. The effects of gender, age, menopausal status, duration of disease and medications on serum allantoin and uric acid levels of the patient and control groups were studied. Our results suggest that uric acid acts as a free radical scavenger and thus is converted to allantoin. Increased allantoin levels suggest the possible involvement of free radicals in rheumatoid arthritis. PMID- 15346654 TI - NADPH oxidase-mediated generation of reactive oxygen species is critically required for survival of undifferentiated human promyelocytic leukemia cell line HL-60. AB - Nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) mediated generation of reactive oxygen species (ROS) was originally identified as the powerful host defense machinery against microorganism in phagocytes. But recent reports indicated that some non-phagocytic cells also have the NADPH oxidase activity, and the ROS produced by it may act as cell signal molecule. But as far as today, whether the NADPH oxidase also plays similar role in phagocyte has not been paid much attention. Utilizing the undifferentiated HL-60 promyelocytic leukemia cells as a model, the aim of the present study was to determine whether NADPH oxidase plays a role on ROS generation in undifferentiated HL-60, and the ROS mediated by it was essential for cell's survival. For the first time, we verified that the release of ROS in undifferentiated HL-60 was significantly increased by the stimulation with Calcium ionophore or opsonized zymosan, which are known to trigger respiration burst in phagocytes by NADPH oxidase pathway. Diphenylene iodonium (DPI) or apocynin (APO), two inhibitors of NADPH oxidase, significantly suppressed the increasing of ROS caused by opsonized zymosan. Cell survival assay and fluorescence double dyeing with acridine orange and ethidium bromide showed that DPI and APO, as well as superoxide dismutase (SOD) and catalase (CAT) concentration-dependently decreased the viability of undifferentiated HL-60 cells, whereas exogenous H2O2 can rescue the cells from death obviously. Our results suggested that the ROS, generated by NADPH oxidase play an essential role in the survival of undifferentiated HL-60 cells. PMID- 15346655 TI - The effect of catechol O-methylation on radical scavenging characteristics of quercetin and luteolin--a mechanistic insight. AB - The biological effect of flavonoids can be modulated in vivo due to metabolism. The O-methylation of the catechol group in the molecule by catechol O-methyl transferase is one of the important metabolic pathways of flavonoids. In the present study, the consequences of catechol O-methylation for the pH-dependent radical scavenging properties of quercetin and luteolin were characterized both experimentally and theoretically. Comparison of the pKa values to the pH dependent TEAC profiles reveals that O-methylation not only affects the TEAC as such but also modulates the effect of changing pH on this radical scavenging activity due to an effect on the pKa for deprotonation. The pH-dependent TEAC curves and computer calculated electronic parameters: bond dissociation energy (BDE) and ionisation potential (IP) even indicate that O-methylation of the luteolin catechol group affects the radical scavenging potential only because it shifts the pKa for deprotonation. O-Methylation of the quercetin catechol moiety affects radical scavenging capacity by both an effect on the pKa, and also by an effect on the electron and hydrogen atom donating properties of the neutral (N) and the anionic (A) form of the molecule. Moreover, O-methylation of a catechol OH-group in quercetin and luteolin has a similar effect on their TEAC profiles and on calculated parameters as replacement of the OH-group by a hydrogen atom. Altogether, the results presented provide new mechanistic insight in the effect of catechol O-methylation on the radical scavenging characteristics of quercetin and luteolin. PMID- 15346656 TI - Promoting effects of polyunsaturated fatty acids on chromosomal giant DNA fragmentation associated with cell death induced by glutathione depletion. AB - Glutamate and buthionine sulfoximine (BSO) both reduce intracellular glutathione (GSH) concentration but by different mechanisms, and thereby induce cell death in C6 rat glioma cells. The effects of lipid peroxidation on chromosomal DNA damage during the GSH depletion-induced cell death were assessed. Polyunsaturated fatty acids (PUFA), such as arachidonic acid (AA), gamma-linolenic acid and linoleic acid enhanced lipid peroxidation, induced a loss of membrane integrity and consequently promoted 1-2 Mbp giant DNA fragmentation under both glutamate- and BSO-induced GSH-depletion. Treated C6 cells had 3'-OH termini in their DNA which were recognized by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) analysis. Antioxidants capable of scavenging reactive oxygen species and lipid radicals and iron or copper scavengers inhibited both lipid peroxidation and 1-2 Mbp giant DNA fragmentation, consequently protecting against cell death under GSH depletion. These results suggest that GSH depletion induces lipid peroxidation and leads to 1-2 Mbp giant DNA fragmentation; and that PUFAs can promote giant DNA fragmentation and 3'-OH termini in chromosomal DNA enhancing lipid peroxidation of C6 cells. PMID- 15346657 TI - Oxidation products and antioxidant markers in plasma of patients with Graves' disease and toxic multinodular goiter: effect of methimazole treatment. AB - Oxidative stress plays an important role in hyperthyroidism-induced tissue damage, as well as in development of autoimmune disorders. To clarify influence of thyroid metabolic status and autoimmune factors on blood extracellular indices of reactive oxygen species (ROS) generation and free radical scavenging in hyperthyroidism, we studied patients with newly diagnosed and untreated Graves' disease without infiltrative ophthalmopathy (17 female and 8 male, aged 41.8 +/- 8.9) and toxic multinodular goiter (15 female and 9 male, aged 48.4 +/- 10.1) under the same antithyroid treatment protocol. Initially and after achievement of stable euthyroidism with methimazole, plasma levels of hydrogen peroxide (H202), lipid hydroperoxides (ROOH) and ceruloplasmin (CP) and serum concentrations of thiobarbituric acid-reacting substances (TBARS) were determined. Similarly, activities of plasma superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) were assayed. The results were compared to those of age- and sex-matched controls. Average duration of hyperthyroidism and treatment period were similar in both patients groups. H202, ROOH and TBARS concentrations were significantly higher in hyperthyroid patients compared to controls. Hyperthyroidism caused an evident increase in SOD and CAT activities and CP level, as well as a decrease in GPx and GR activities. Achievement of euthyroidism resulted in normalization of all analyzed parameters in both hyperthyroid patients groups. These findings suggest that the changes in blood extracellular indices of oxidative stress and free radical scavenging in hyperthyroid patients are influenced by thyroid metabolic status, and are not directly dependent on autoimmune factors present in Graves' disease. PMID- 15346658 TI - Evaluation of the effect of tibolone and transdermal estradiol on triglyceride level in hypertriglyceridemic and normotriglyceridemic postmenopausal women. AB - The objective of the present study was to quantify the magnitude of the association between the change in triglycerides and high-density lipoprotein (HDL) cholesterol levels with the use of tibolone and transdermal estrogen in postmenopausal women with hypertriglyceridemia and normotriglyceridemia. This prospective randomized study enrolled 140 postmenopausal women who had all been hysterectomized for almost a year or more. All subjects completed the 3-month follow-up. The 140 patients were divided into two groups: 70 were given transdermal 17beta-estradiol 0.05 mg/day, and 70 were given tibolone 2.5 mg/day. We compared the effects of tibolone and transdermal 17beta-estradiol on lipids and dimacteric symptoms of the patients. To evaluate the effects of tibolone and transdermal estrogens on triglycerides and HDL cholesterol in postmenopausal women with normotriglyceridemia and hypertriglyceridemia, the women were assigned to five groups according to triglyceride levels (0-100, 101-200, 201-300, 301-400 and > or = 401 mg/dl). We compared changes in the triglyceride and HDL cholesterol levels of each group after treatment. All 140 postmenopausal women completed the trial. No significant differences were found in baseline characteristics of the patients. The tibolone group showed a 22.6% decrease whereas the transdermal estrogen group had a 10.9% decrease in the mean triglyceride levels after 3 months of treatment. The mean decrease of triglyceride level with transdermal estradiol was approximately 11% in normotriglyceridemic and hypertriglyceridemic postmenopausal women. The mean decrease of triglyceride level was 17% in the normotriglyceridemic group and 22 30% in the hypertriglyceridemic groups with tibolone. While the mean HDL cholesterol level increased in the transdermal estrogen group (3.6%), it decreased in the tibolone group (9.3%). We found that tibolone decreased triglyceride levels much more than did transdermal estradiol. However, HDL cholesterol was decreased by tibolone and increased by transdermal estradiol. Tibolone had a more marked decreasing effect in postmenopausal women who had higher initial triglyceride levels. It is suggested that the beneficial effect of tibolone on the cardiovascular system might be greater in women with a high level of triglycerides. PMID- 15346659 TI - High local endometrial effect of vaginal progesterone gel. AB - Our objective was to investigate the first-pass effect to the uterus of progesterone gel administered vaginally. This was a prospective, randomized study of 32 postmenopausal women, attending our menopause clinic. All women used transdermal estradiol (50 microg/day, a patch each week) for 2 weeks. They used either vaginal progesterone gel or intramuscular progesterone in oil 50 mg after 24 h to oppose the transdermal estradiol. Serum progesterone levels and endometrial tissue progesterone levels were determined. Serum progesterone levels were higher in women who used the intramuscular rather than the vaginal route. Although serum progesterone levels in the vaginal group were lower than in the intramuscular group, the endometrial tissue concentration of progesterone was higher. It is concluded that progesterone gel, used vaginally, has a high local effect on the endometrium, without any systemic side-effects due to high plasma progesterone levels. PMID- 15346661 TI - Ultrasound measurement of pregnancy-induced changes in maternal bone mass: a longitudinal, cross-sectional and biochemical study. AB - Our objective was to investigate the changes in maternal bone status in pregnancy. Amplitude-dependent speed of sound (AD-SoS) by quantitative ultrasound was measured in 100 healthy, White, pregnant women at term, and 100 White, healthy, non-pregnant and premenopausal women matched for age, as controls. In a subgroup of 50 women a longitudinal evaluation throughout pregnancy was performed. A significant reduction of AD-SoS was observed during pregnancy (controls: 2170 +/- 55 m/s; first trimester: 2118 +/- 46 m/s; second trimester: 2085 +/- 49 m/s; third trimester: 2081 +/- 51 m/s), showing a negative correlation with body mass index (r -0.31; p < 0.05) and a positive correlation with daily calcium intake (r 0.33; p < 0.05). Increased levels of urinary markers of bone resorption confirmed bone turnover (p < 0.05). Ultrasonographic study of bone is a simple, low-cost and safe method for measuring maternal bone mass in pregnancy. During pregnancy, there was a significant loss in AD-SoS that is an indicator of bone status; this decrease was higher in the second and third trimesters (p < 0.05), associated with a high bone turnover. It was more intense in women with a low calcium intake (p < 0.05). PMID- 15346660 TI - Effect of hormone replacement therapy, tibolone and raloxifene on serum lipids, apolipoprotein A1, apolipoprotein B and lipoprotein(a) in Greek postmenopausal women. AB - The aim of this study was to assess the effect of estrogen, two regimens of continuous combined hormone replacement therapy (HRT), tibolone and raloxffene on serum lipid, apolipoprotein A1 and B and lipoprotein(a) levels in Greek postmenopausal women. A total of 350 postmenopausal women were studied in a prospective open design. Women were assigned to one of the following regimens depending on the presence of risk factors for osteoporosis, dimacteric symptoms and an intact uterus: conjugated equine estrogen 0.625 mg (CEE, n = 34), continuous combined CEE 0.625 mg plus medroxyprogesterone acetate (MPA) 5 mg, (n = 80), continuous combined 17beta-estradiol 2 mg plus norethisterone acetate (NETA) 1 mg (n = 58), tibolone 2.5 mg (n = 83) and raloxifene HCl 60 mg (n = 50). Forty-five postmenopausal women with no indications for HRT served as controls. Total cholesterol (TC), low-density lipoprotein (LDL) cholestrol and high-density lipoprotein (HDL) cholesterol, triglyceride (TG), apolipoprotein A1 (ApoA1), apolipoprotein B (ApoB) and lipoprotein(a) (Lp(a)) levels were assessed in each subject at baseline, and at 6 and 12 months of therapy. All therapy regimens lowered TC levels compared to baseline (4.2-8.0% decrease). This effect was more prominent in the subgoup of women with high baseline TC levels (9.1-20.4% decrease). LDL cholesterol decreased significantly in CEE, CEE/MPA and raloxifene groups (-11.2%, -11.9% and -11.0%, respectively). Hypercholesterolemic women exhibited a steeper decrease in LDL cholesterol (10.6-27.8% in all therapy groups). TG levels increased significantly in the CEE and CEE/MPA groups (23.7% and 21.8%, respectively), while estradiol/NETA had no effect on TG levels. Tibolone decreased TG levels markedly, by 20.6%, while raloxifene had no TG lowering effect. HDL cholesterol and ApoA1 were increased by CEE and CEE/MPA (HDL cholesterol, 7.4% and 11.8%, respectively; ApoA1, 17.8% and 7.9%, respectively) and decreased by tibolone (HDL cholesterol, -13.6%; and ApoA1, -9.9%). All therapy regimens except raloxifene lowered Lp(a) levels, with tibolone having the more pronounced effect (-13.2 to -29.0%). In conclusion, each therapy regimen had a diferent effect on lipid-lipoprotein levels, exerting favorable and unfavorable modifications. Hypercholesterolemic women seemed to benefit more from the cholesterol-lowering effect of estrogen replacement therapy/HRT. The choice for a particular regimen should be based on individual needs, indications and lipid lipoprotein profile. PMID- 15346662 TI - Augmentation of leptin and hypoxia-inducible factor 1alpha mRNAs in the pre eclamptic placenta. AB - The placenta is a major source of leptin in the fetomaternal circulation, although its physiological role remains to be clarified. Leptin in the fetomaternal circulation is proposed to be a marker of acute stress in the fetus, and the fetus suffering from pre-eclampsia would be under chronic stress. In 16 pre-eclamptic placentas, the expressions of leptin, hypoxia-inducible factor 1alpha (HIF1alpha) and leptin receptor mRNAs were analyzed by semi-quantitative reverse-transcriptase-polymerase chain reaction and compared with clinical data. The co-expressions of leptin and the isoforms of the leptin receptor were observed in all the pre-eclamptic placentas. Leptin mRNA was significantly augmented in the pre-eclamptic placentas, although the level in fetal plasma was not high. The level of the expression of leptin mRNA was correlated with the placental HIF1alpha mRNA level and fetal body weight, but not with the levels of the leptin receptor isoforms in the pre-eclamptic placentas. This observation may suggest that autocrine/paracrine regulation of leptin exists in the human placenta and is upregulated in the pre-eclamptic placenta. PMID- 15346663 TI - The human chorionic gonadotropin molecule from patients with trophoblastic diseases has a high thyrotropic activity but is less active in the ovary. AB - To examine the pathogenesis of hyperthyroidism in women with trophoblastic diseases, the biological activity of human chorionic gonadotropin (hCG) molecules in women with normal pregnancy (n = 85) and in women with trophoblastic diseases (vesicular mole, n = 30; and choriocarcinoma, n = 12) was compared. Hyperthyroidism (thyroid stimulating hormone (TSH) < 0.3 mIU/l) was observed more frequently in women with trophoblastic diseases. All the sera were then subjected to Chinese hamster ovary cells transfected with the human TSH receptor (CHO-hTSHr cells) and cAMP production was compared. Sera from the women with choriocarcinoma showed the highest cAMP production. Interestingly, significant correlation between serum hCG level and cAMP production in CHO-hTSHr cells was observed only in women with trophoblastic disease. All the sera were then applied to CHO cells transfected with hCG/luteinizing hormone (LH) receptor (CHO-hCG/LHr cells). In contrast to the findings with the TSH receptor, sera from the women with normal pregnancy showed the highest cAMP production in these cells. Correlation between serum hCG level and cAMP production in CHO-hCG/LHr cells was significant only in normal pregnancy. These results indicate that the hCG molecule from women with trophoblastic diseases displays enhanced thyrotropic activity. PMID- 15346664 TI - Present concept of unexplained infertility. AB - Despite improvements in both diagnostic assessment and treatment of infertile couples, many couples still have no explanation for their infertility. Unexplained infertility (the failure to conceive of a couple in whom no definitive cause for infertility can be found) has an incidence of 10-20% in all infertile couples. The incidence varies with the population studied and with the criteria used. Unexplained infertility is not an absolute condition but rather a relative inability to conceive, and many of these couples may conceive without treatment. The treatment options for unexplained infertility are several and the treatment results are promising. Expectant management can be recommended if the woman is under 28-30 years of age and the infertility duration is less than 2-3 years. In vitro fertilization (IVF) has revolutionized the treatment of infertile couples, as well as profoundly increasing the basic understanding of human reproduction. IVF can be used as both a diagnostic and a therapeutic tool in couples with unexplained infertility. The pregnancy rates with IVF are good, at 40% per treatment cycle. In addition, the outcome of pregnancies among women with unexplained infertility is generally comparable to that of spontaneous and other pregnancies using assisted reproductive technologies. PMID- 15346666 TI - Postpartum depression beyond the early postpartum period. AB - OBJECTIVE: To review the literature concerning maternal postpartum depression beyond the early postpartum period. DATA SOURCES: A literature search was conducted using Cinahl, Medline, and PsychInfo electronic databases. Keywords included postpartum depression, postpartum depressive symptoms, course, prevalence, incidence, and remission. STUDY SELECTION: Studies selected included incidence of maternal depression or depressive symptoms, existing in the early postpartum period, and measured again at postpartum points from 6 months through 2 1/2 years after delivery. Only studies published in English were included. Twenty-three articles were located, and a recent relevant study conducted by the current author also was included. DATA EXTRACTION: Studies were reviewed and data organized according to year, sample characteristics, time of depression assessment, instrument used, cutoff score, rate of depression, and factors associated with depression at later postpartum time points. DATA SYNTHESIS: For a significant percentage of women, postpartum depressive symptoms continue for months or even years after giving birth. Factors associated with postpartum depression at later time points are identified. CONCLUSIONS: Continued evaluation of women with elevated depression levels at initial screening, and treatment for women whose depression does not remit spontaneously within the first few weeks or months postpartum is recommended. Further research is needed to understand the phenomenon of persistent postpartum depression. PMID- 15346667 TI - Family-centered care: do we practice what we preach? AB - OBJECTIVE: To determine nurses' perceptions and practices of identified elements of family-centered care. DESIGN: Descriptive. SETTING: Neonatal intensive care unit (NICU), pediatrics, and pediatric intensive care unit (PICU) in an acute care hospital. PARTICIPANTS: Sixty-two licensed nurses, 37 working in the NICU and 25 working in pediatrics or the PICU. MAIN OUTCOME MEASURES: Scores for the Necessary and Current scales of the Family-Centered Care Questionnaire. RESULTS: Scores representing current nursing practice of family-centered care were significantly lower than those representing its necessity (p = .000). Nurses with 10 years or fewer of neonatal or pediatric experience scored significantly higher on both the total Necessary Scale (p = .02) and total Current Scale (p = .017) than did those with 11 years or more. Nurses who work in the NICU scored significantly lower on the total Necessary Scale (p = .013) than did nurses who work in pediatrics or PICU. CONCLUSIONS: Although nurses agree the identified elements of family-centered care are necessary, they do not consistently apply those elements in their everyday practice. Years of experience and clinical work setting influenced both perceptions and practices of family-centered care. PMID- 15346668 TI - Barriers to prenatal care for homeless pregnant women. AB - OBJECTIVE: The purpose of this study was to explore and describe the barriers to prenatal care for homeless pregnant women. DESIGN: A descriptive survey with written questionnaires. PARTICIPANTS: About 183 pregnant homeless women live in Northeast Florida in any given month. More than 250 surveys were distributed to homeless pregnant women via agencies that provide shelter and/or services to homeless people in Northeast Florida. Forty-seven surveys were returned, representing 25.7% of the estimated population. MAIN OUTCOME MEASURE: Barriers to prenatal care were measured using Melnyk's Barriers Scale, a 27-item Likert-type scale. RESULTS: Of the respondents, 75.61% perceived barriers to prenatal care. Site-related factors were the most significant, followed by provider/client relationship, inconvenience, fear, and cost. CONCLUSIONS: Although prenatal care is available to all pregnant women in Northeast Florida, barriers to accessing this care for homeless pregnant women are significant and need to be addressed by health care providers and administrators. Additional qualitative study of these issues is warranted. PMID- 15346669 TI - Predictors of prenatal attachment in mothers of twins. AB - OBJECTIVES: To explore the predictive relationship between prenatal attachment and selected demographic and biopsychosocial factors and to compare these predictors to those identified in previous research on maternal attachment in singleton pregnancies. DESIGN: A predictive correlational descriptive design. SETTING: Data were obtained via mailed surveys. PARTICIPANTS: 214 women expecting twins were recruited from a national mother of twins support group. MAIN OUTCOME MEASURES: Instruments included the Rosenberg Self-Esteem Scale, Norwood Social Support Apgar, Miller Prenatal Attachment Inventory, and a demographic data tool to collect data on perinatal factors of gestational age, infertility history, perceived risk status, fetal movement, and planning of pregnancy. RESULTS: Women who were younger, with lower income, a history of infertility, greater self esteem, who had experienced quickening, and were further along in their pregnancy reported greater prenatal attachment to their twins (Adjusted R2 = 19.4%, p < .001). Perceived risk, social support, and planning of pregnancy did not predict level of attachment. CONCLUSION: Factors that influence attachment in women experiencing a twin pregnancy are complex, and much is still unknown. Development and testing of theory is needed to guide future research and practice. PMID- 15346670 TI - The utility of fFN for the prediction of preterm birth in twin gestations. AB - OBJECTIVE: To determine the sensitivity, specificity, and positive predictive value (PPV) of fetal fibronectin (fFN) and to determine its usefulness, in conjunction with selected other clinical assessment measures, in the prediction of preterm birth for women with twin gestations. DESIGN: A prospective, descriptive, longitudinal design. SETTING: An obstetrical high-risk clinic that received patient referrals from several surrounding communities in central Texas. PATIENTS/PARTICIPANTS: Forty-eight women identified with twin gestations prior to the 22nd week of pregnancy; primarily of Hispanic ethnicity. MAIN OUTCOME MEASURES: A substantial number of outcome variables were assessed in this study. In the present report, data derived from weekly assessments for the identification of the presence of fFN, the diagnosis of bacterial vaginosis, and the measurement of cervical length were reviewed for their relationship to prematurity, birth weight, birth weight discordancy and placental chorionicity. RESULTS: The relative risk of birth prior to 35 weeks gestation, fetal death, or discordance of twin birth weights of greater than 20% was 2.22 (CI: 1.09, 4.55, P < 0.015) when fFN was found to be positive at any weekly testing after 22 to 24 weeks gestation (sensitivity 76.82%, specificity 58.33%, PPV 66.7%). The presence of fFN was most highly predictive of preterm birth when performed during the 24th to 28th gestational week. Shorter cervical lengths were highly correlated with preterm birth (r = -0.6). An association between bacterial vaginosis and preterm birth was not demonstrated in this sample. CONCLUSION: Sampling for the presence of fetal fibronectin can be easily accomplished by RNs in labor triage units and by advanced practice nurses in outpatient settings. The identification of fFN, particularly during the 24 to 28 weeks gestational time frame, is highly predictive of preterm birth, and particularly so for women with twin gestations. PMID- 15346671 TI - Achieving a new balance: women's transition to second-time parenthood. AB - OBJECTIVE: To describe the experience of transition to second-time parenthood from the mother's perspective. DESIGN: Descriptive phenomenology using Colaizzi's approach. SETTING: Participants were interviewed in their homes. PARTICIPANTS: Ten second-time mothers with a second child between the ages of 6 months and 24 months. RESULTS: The birth of the second child motivated these women to seek and achieve a new balance to integrate a second child into the existing family. Seven themes emerged as common elements in all of the women's lives: (a) balancing the positive and negative elements of the early weeks, (b) knowing what to expect, (c) establishing a new routine, (d) maintaining the marital relationship, (e) taking a break, (f) seeking out support, and (g) nurturing relationships among family members. CONCLUSIONS: Mothers overwhelmingly found the addition of a second child to the family to be a positive experience regardless of the effort it took to accommodate their lives to the needs of two children. Although second time mothers approach the experience with a wealth of knowledge, this research highlights that these women may have concerns that are not addressed. The nurse can focus assessments and offer interventions to meet the needs of these women as they become mothers for the second time. PMID- 15346672 TI - Maternal perspectives on postpartum return to the workplace. AB - OBJECTIVE: To describe maternal perceptions about postpartum return to the workplace. DESIGN AND SETTING: The research framework was McCubbin and McCubbin's Resiliency Model of Family Stress, Adjustment, and Adaptation. Employed, postpartum women, who resided in six different states, were recruited from day care and primary care settings to participate in this descriptive study. PARTICIPANTS: Seventy-four employed, married women, who returned to the workplace within one year after childbirth. MAIN OUTCOME MEASURE: As part of a larger study, perceptions about combining multiple roles after return to the workplace are the focus for this report. RESULTS: Participants responded to open-ended questions about their return to the workplace during the first months after giving birth. Content analysis of those responses resulted in two major categories, Resiliency Challenges: Negative Aspects (role conflict/overload, family stress, family/child issues, finances, psychosocial issues) and Resiliency Building: Positive Aspects (social support, maternal role satisfaction, positive adaptation, career role satisfaction). CONCLUSIONS: The data support the premise that employed women view returning to the workforce as having more challenges than they expected because the experience was viewed as being mostly negative. Preparing for return to the workplace is an important, but neglected, topic that needs to be addressed and defined more clearly in the literature. Therefore, future research is needed to identify resources and interventions that will help women experience fewer challenges associated with postpartum employment. PMID- 15346673 TI - Nurses' attitudes toward pregnancy termination in the labor and delivery setting. AB - OBJECTIVE: To examine nurses' attitudes toward pregnancy termination in the labor and delivery setting and the frequency of nurse refusal to care for patients undergoing pregnancy termination. DESIGN: Nonexperimental, descriptive study. SETTING: Six central and northern California hospitals, including Level 1, 2, and 3 facilities. PARTICIPANTS: Seventy-five labor and delivery registered nurses. METHOD: Anonymous survey with visual analog scales. RESULTS: Ninety-five percent of the nurses indicated they would agree to care for patients terminating a pregnancy because of fetal demise, 77% would care for patients terminating a fetus with anomalies that were incompatible with life, and 37% would care for patients terminating for serious but nonlethal anomalies, with a significant drop in agreement as gestation advanced. Few nurses would agree to care for patients undergoing termination for sex selection, selective reduction, or personal reasons. Nurses both accepting and refusing patient care assignments were criticized by coworkers. CONCLUSION: Clear guidelines should be established on how to handle nurse refusal to care for patients terminating pregnancy in advance. Open discussions should be encouraged between staff and management to minimize criticism. PMID- 15346674 TI - Survivors of childhood sexual abuse: implications for perinatal nursing care. AB - Childhood sexual abuse has a lifelong impact on its survivors and may affect the childbearing experience in a variety of ways. Nurses caring for women during the perinatal period can benefit from understanding the phenomenon of child sexual abuse, its prevalence, and the sequelae experienced by childbearing women. Signs, symptoms, and correlates of prior sexual abuse manifested during the perinatal period are described, and nursing care responses are suggested. Nurses may be able to use this information to recognize survivors and facilitate both a positive childbirth experience and the healing process, by helping the survivor learn to trust, respect, and care for her body. PMID- 15346675 TI - Childbearing women's perceptions of nursing care that promotes dignity. AB - OBJECTIVE: To gain an understanding of the perceptions of childbearing women about the maintenance of dignity while laboring and giving birth. DESIGN: Descriptive qualitative study. SETTING: A university community in the western United States. PATIENTS/PARTICIPANTS: Twenty low-risk primiparous women who had recently given birth to healthy term neonates. MAIN OUTCOME MEASURES: Semistructured audio-taped interviews were conducted in the homes of participants using an interview guide. RESULTS: The following themes were identified: (a) nurses played a pivotal role in preserving dignity during childbirth, (b) women appreciated feeling valued and respected, and (c) dignity was enhanced by nursing care that gave women their preferred level of control. CONCLUSION: Nursing behaviors that demonstrate valuing and respect of childbearing women are essential in preserving the quality of the birth experience. PMID- 15346676 TI - Making meaning: women's birth narratives. AB - Birth stories are personal narratives grounded in the pivotal life experience of giving birth. Richly descriptive birth narratives from culturally diverse childbearing women document the importance of listening to the voices of women. Benefits of sharing birth stories include the opportunity for integration of a major event into the framework of a mother's life; the opportunity to share a significant life experience; the opportunity to discuss fears, concerns, "missing pieces" or feelings of inadequacy or disappointment; the opportunity for the woman to gain an understanding of her strengths; and the opportunity to connect with other women. Providing women with the opportunity to share their birth stories is an important nursing intervention. PMID- 15346677 TI - Is globalization reducing poverty and inequality? AB - Over the past 20 years or so, India, China, and the rest of East Asia experienced fast economic growth and falls in the poverty rate, Latin America stagnated, and the former Soviet Union, Central and Eastern Europe, and sub-Saharan Africa regressed. But what are the net trends? The neoliberal argument says that world poverty and income inequality fell over the past two decades for the first time in more than a century and a half, thanks to the rising density of economic integration across national borders. The evidence therefore confirms that globalization in the context of the world economic regime in place since the end of Bretton Woods generates more "mutual benefit" than "conflicting interests." This article questions the empirical basis of the neoliberal argument. PMID- 15346678 TI - Social austerity versus structural reform in European health systems: a four country comparison of health reforms. AB - Cost containment has captured the attention of health policymakers in most OECD countries, and deliberations about creating powerful financial incentives dominate health care politics. Some European health systems are now implementing hospital payment schemes that mirror the U.S. model of diagnosis-related groups (DRGs) and are raising premiums and copayment levels in an effort to limit public expenditures. Though financial incentives may indeed help rein in health expenditures, focusing predominantly on financial incentives hinders due consideration of needed structural reforms that improve the continuity, quality, and appropriateness of health care service delivery. This article focuses on the structural specifics of two legally enacted health insurance systems (Germany and Austria) and two national health systems (Great Britain and Denmark) to discuss the influence of structural characteristics on cost-containment efforts. Structural reform strategies discussed include increasing reliance on general practitioners, improving coordination of community and hospital-based specialty care, addressing the stark divide between ambulatory and hospital-based care that exists in some European health systems, and improving continuity of care by better integrating medical and social care sectors. Also discussed is the relative focus on financial incentives versus structural deficits in recent European-health care reform strategies. PMID- 15346679 TI - Public health sector unions and deregulation in Europe. AB - Deregulation and liberalization of health services take several forms in Europe: public-private partnerships; contracting out of services; and corporatization of health care institutions. The impact on health workers includes changes in terms and conditions of employment, breakup of collective bargaining agreements, and often more stressful working conditions. The author examines four types of trade union responses to deregulation. National health trade union action has used campaigning, awareness raising, and judicial review. Health workers' unions in alliance with other trade unions have taken part in wider campaigns against privatization and in promoting public services. Health workers' unions joining with social movements have become involved in wider alliances that link with broader public policy issues such as poverty reduction and urban/regional regeneration. European-wide action, seen through the work of the European Federation of Public Service Unions, has concentrated on the development of an alternative health policy, and the promotion of social dialogue at a European level. Trade unions must adopt a range of approaches to challenge the effects of deregulation. Increasingly, trade union members need to be aware of how to take action at both the national and European levels. PMID- 15346680 TI - Developing countries and trade in health services: which way is forward? AB - International trade in health services appears to be increasing It may receive a further boost when liberalized and bound under international trade agreements, such as the General Agreement on Trade in Services (GATS). Liberalization of trade in health services can create opportunities, but may also exacerbate preexisting problems. Moreover, once liberalization is locked in under international trade agreements, reversing policies becomes difficult, especially for developing countries. Making undue commitments to liberalize health services under GATS may therefore result in the loss of policy space. Yet the GATS agreement contains considerable flexibility to fine-tune commitments in accordance with national (health) objectives. But flexibility entails complexity, and for GATS the complexity is compounded by the fact that some of its rules are still being developed. Moreover, flexibility is meaningless unless used, and used well, which calls for a profound analysis of the alternatives and their implications. This article provides an overview of considerations related to (international) trade in health services and key features of GATS that are relevant to public health. It highlights that policymakers have something at stake in GATS negotiations; thus, they should take an interest in liberalization of trade in health services, analyze its implications, and give input and guidance to their country's trade negotiators. Moreover, to convey their concerns and aspirations effectively, they must learn the trade language, while staying focused on (public) health objectives. PMID- 15346681 TI - International trade agreements: hazards to health? AB - Since the 1980s, neoliberal policies have prescribed reducing the role of governments, relying on market forces to organize and provide health care and other vital human services. In this context, international trade agreements increasingly serve as mechanisms to enforce the privatization, deregulation, and decentralization of health care and other services, with important implications for democracy as well as for health. Critics contend that social austerity and "free" trade agreements contribute to the rise in global poverty and economic inequality and instability, and therefore to increased preventable illness and death. Under new agreements through the World Trade Organization that cover vital human services such as health care, water, education, and energy, unaccountable, secret trade tribunals could overrule decisions by democratically elected officials on public financing for national health care systems, licensing and training standards for health professionals, patient safety and quality regulations, occupational safety and health, control of hazardous substances such as tobacco and alcohol, the environment, and affordable access to safe water and sanitation. International negotiations in 2003 in Cancun and in Miami suggested that countervailing views are developing momentum. A concerned health care community has begun to call for a moratorium on trade negotiations on health care and water, and to reinvigorate an alternative vision of universal access to vital services. PMID- 15346682 TI - Heritable anomalies among the inhabitants of regions of normal and high background radiation in Kerala: results of a cohort study, 1988-1994. AB - In a genetic epidemiological and fertility survey among 70,000 inhabitants in a high-background radiation region (HBRR) and normal radiation region (NRR) in Kerala, India, 985 persons were found to have heritable anomalies. Suggested etiologies for the anomalies were chromosomal and Mendelian, 15 percent; multifactorial, 60 percent; and congenital, 25 percent. There was a statistically significant increase of Down syndrome, autosomal dominant anomalies, and multifactorial diseases and an insignificant increase of autosomal recessive and X-linked recessive anomalies in the HBRR. The total fertility rate was 3.85 per couple; 9 percent of live-born children were reported dead. The rate of untoward pregnancy outcome--death of the offspring or presence of an anomaly in a living child--was 6.4 percent among the unrelated couples in the NRR, with one spouse born outside the area of current residence ("migrant"). Considering this as the base, the excess relative risks in the other groups are: "NRR-nonmigrant," 35 percent; "HBRR-nonmigrant," 69 percent; "NRR-consanguineous," 76 percent; and "NBRR-consanguineous," 157 percent. Ionizing radiation, consanguinity, and nearness of birthplace of the spouse are risk factors for the death of offspring and for anomalies. The higher risk among the "nonmigrant" couples may be due to geographic inbreeding. The findings are suggestive of an autosomal recessive etiology for the majority of the multifactorial anomalies. PMID- 15346683 TI - A free market solution for prescription drug crises. AB - The cost of prescription drugs is imposing an ever greater burden on families and varying levels of government. The vast majority of this cost is attributable to patent protection, since most drugs are actually relatively cheap to produce. The temporary monopolies provided by patent protection have been the main mechanism through which corporations have financed their drug research. This article examines the efficiency of publicly supported drug research relative to the current patent system. The author shows that even if publicly funded research were considerably less efficient on a dollar-per-dollar basis than patent supported research, there would still be enormous gains from switching to a system of publicly supported research. The main reason for this conclusion is that patent monopolies lead to enormous economic distortions, including expensive sales promotion efforts, research into "copycat drugs," incentives to conceal unfavorable research findings, and other inefficiencies that economic theory predicts would result from a government-created monopoly. The gains from publicly supported research, coupled with a free market in the production of drugs, could reach into several hundred billion dollars annually within a decade. PMID- 15346684 TI - Transnational capital and confessional politics: the paradox of the health care system in Lebanon. AB - For the past two decades there has been a debate over the implementation of structural adjustment policies in the health sector of developing countries, much of it focusing on the political and economic relevance of the reform process for public health provision. However, very few studies have been able to assess the relevance of the private sector, which has had a central role in the restructuring of health services worldwide. Lebanon provides just such a case, with a predominantly private provider and the role of the state relegated to financing, with few controls over supply. This situation has ensured the systematic destruction of what remained of public provision in the 1970s. The country is now faced with one of the most expensive health services in the world, and one in which much of the population continues to live under conditions of considerable economic deprivation. The unique situation of Lebanon is maintained by its politics of confessionalism, with sociopolitical relations dominated by primordial ties of family, tribe, and kin, which does not seem to be an obstacle to the process of globalization. The authors suggest that this context reinforces the gross inequalities in access to health care; they explore the complex relationship between state, finance capital, and confessional politics in the context of health sector reform, in particular the financing of health care. PMID- 15346685 TI - United States accused of sabotaging obesity strategy. AB - In early 2004, the U.S. Department of Health and Human Services disputed some of the scientific evidence underlying the World Health Organization's proposals for curbing obesity. Critics of the DHHS position suggest that it puts the interests of the junk food industry ahead of people's health. PMID- 15346686 TI - Clinical trials seen shifting overseas. AB - A Tufts University study released in mid-2003 indicates that increasing numbers of clinical trials for U.S.-produced pharmaceutical are being conducted outside the United States, mainly by contract research organizations. This process speeds up clinical trials, but raises ethical issues of the possible coercion or exploitation of more vulnerable and naive patient populations. PMID- 15346687 TI - Specter orange. AB - Nearly 30 years after the Vietnam War, a chemical weapon used by U.S. troops is still exacting a hideous toll on each new generation in Vietnam. The dioxin (TCCD) that contaminated the herbicide Agent Orange is one of the most toxic molecules known to science. The contaminant persists in the soil. The United States has done nothing to combat the medical and environmental catastrophe that is overwhelming the country. PMID- 15346688 TI - Health of prisoners, and the public health: no man is an island. PMID- 15346689 TI - The carceral limb of the public body: jail inmates, prisoners, and infectious disease. PMID- 15346690 TI - Actual medication pill burden and dosing frequency in HIV-infected patients with undetectable viral loads. AB - Studies have shown equivalent or improved outcomes with once-daily antiretroviral (ARV) regimens, but have failed to quantify the true daily dosing frequency and pill burden of HIV-infected patients, by not considering concomitant medications. This study was conducted to describe the total daily number (pill burden) and dosing frequency for all oral medicines, and not just ARV medications in HIV infected patients with undetectable viral loads. A retrospective chart review of patients with undetectable viral loads in an HIV primary care clinic identified dosing frequency and pill burden for all prescribed medications. These patients were divided into those with concomitant medications (WITH) and those only on ARV medications (WITHOUT). One hundred and seventeen patients qualified for the evaluation. Ninety-three were in the WITH group and 24 were in the WITHOUT group. WITH patients were older (p < 0.05), and had advanced US Centers for Disease Control and Prevention (CDC)-defined HIV disease stage (p< 0.05), a longer duration of HIV infection (p= 0.03), and a longer duration of AIDS (p= 0.02). WITH patients had a significantly greater total pill burden (p<0.01) and dosing frequency (p<0.001) when considering all medications. Assessing only ARV medications, a significant difference existed only in pill burden (mean +/- SD) (WITH = 9.91 +/- 2.1 versus WITHOUT = 6.6 +/- 1; p < 0.01), and no difference existed in ARV dosing frequency of 2.09 +/- 1.9 and 1.83 +/- 1.7, respectively (p = 0.07). Higher dosing frequencies and pill burdens were present in patients with co-morbid conditions. All patients were on more than once-daily therapies. Dosing requirements for non-ARV medications should also be considered when optimizing or simplifying ARV regimens. PMID- 15346691 TI - Epidemic HIV-associated Kaposi's sarcoma: not as common as it used to be. PMID- 15346692 TI - [The natural history of dilated cardiomyopathy: a review of the Heart Muscle Disease Registry of Trieste]. AB - Dilated cardiomyopathy (DCM), a heart muscle disease characterized by ventricular dilation and dysfunction, is a leading cause of mortality and morbidity. In the present paper we will consider the main results of studies on the natural history of DCM in 581 consecutive patients prospectively enrolled and systematically followed in the Heart Muscle Disease Registry of Trieste in the last 25 years. In the last decades prognosis of DCM significantly improved over time, mainly as a consequence of optimized treatment with ACE-inhibitors and beta-blockers. However, a strong heterogeneity of prognosis was observed among patients both in familial and sporadic cases. Early diagnosis and treatment allowed to recognize two distinct subgroups, one with a rapidly progressive downhill course, high mortality and urgent indication to heart transplantation, another with a more favorable outcome. Long-term optimized treatment with ACE-inhibitors (in 90% of cases) and beta-blockers (in 87% of cases) was associated with a remarkable clinical improvement in 50% of patients and apparent "healing" in 16% of cases. A systematic and accurate echocardiographic follow-up showed in these cases a significant improvement of the left ventricular ejection fraction (LVEF) with "reverse remodeling", frequently associated with a decrease of severity of functional mitral regurgitation and regression of the restrictive filling pattern. The response to optimal treatment showed a strong relation to long-term outcome. The 8-year transplant-free survival, starting from the evaluation at 2 years, was 31% in patients with persistent NYHA class III-IV, 64% in NYHA class I II and LVEF < or = 40%, 83% in NYHA class I-II and LVEF > 40% and 94% in patients with apparent "healing" (p < 0.0001). Long-term follow-up showed a significant clinical progression of the disease in 33% of cases, independently of the initial clinical response to treatment. Predictive factors of a favorable response to beta-blocker treatment associated with ACE-inhibitors were a history of mild hypertension, an early diagnosis and treatment and the presence of sinus tachycardia. The risk of sudden death was increased particularly in patients with long-term persistent or progressive left ventricular dilation and dysfunction. A rigorous pharmacological approach (optimization of beta-blockers, withdrawal or decrease of dosage of digitalis), and selective non-pharmacological strategy (automated implantable cardioverter-defibrillators for primary prevention in high risk patients) are potentially effective to decrease the incidence of sudden death during long-term follow-up. In conclusion, the Heart Muscle Disease Registry of Trieste gave us in the last 25 years new insights into the natural history of DCM, underlying the importance of a rigorous and systematic approach both at clinical presentation and during long-term follow-up on optimized medical treatment. PMID- 15346693 TI - [Equivalence studies in cardiovascular diseases]. AB - An increasing number of equivalence and non-inferiority trials appeared in recent years among randomized clinical trials, particularly for the evaluation of efficacy of treatments in cardiovascular medicine. These studies are characterized by remarkable methodological issues and important ethical implications. The aim of this review was to underscore some topic methodological problems: setting the equivalence boundary, the special logic used to establish equivalence, and the determinants of sample size. A Medline search was performed in order to identify the main problems in study design and in the interpretation of results of the equivalence and non-inferiority trials in cardiovascular diseases. PMID- 15346694 TI - [Off-pump myocardial revascularization in patients with multivessel coronary disease]. AB - BACKGROUND: Some criticisms have been addressed to off-pump coronary surgery technique concerning the possibility of its systematic use with the respect of the completeness of revascularization. We report our experience with off-pump revascularization in patients with multivessel coronary disease. METHODS: Between September 1997 and April 2003, 868 patients with multivessel coronary disease were scheduled for off-pump surgical revascularization. From September 2000, the percentage of patients operated on without cardiopulmonary bypass has been stably > 90%. Fifteen patients (1.7%) had a conversion to cardiopulmonary bypass for anatomical reasons (n = 6) or clinical instability (n = 9). RESULTS: An average of 2.5 +/- 0.8 (range 1-5) anastomoses per patient were completed. Bilateral mammary artery was used in 573 patients (66%); totally arterial revascularization was accomplished in 479 patients (55.2%). In-hospital mortality rate was 0.6% (5 patients). Total incidence of non-fatal postoperative complications (bleeding requiring re-exploration, perioperative myocardial infarction, stroke, new onset of acute renal failure) was 3.5%. Mean postoperative hospital stay was 4.8 +/- 3.8 days. At a mean follow-up of 21.6 +/- 15.6 months (range 1-65 months), the postoperative actuarial survival rates were 97.3, 93.7 and 86.7% at 1, 3 and 5 years postoperatively. Actuarial freedom rates from new revascularization were 98.7, 96.6 and 96.6% at 1, 3 and 5 years postoperatively. CONCLUSIONS: Early- and intermediate-term results of this study demonstrate the feasibility of off-pump revascularization in all patients with multivessel coronary disease, respecting the criterion of complete myocardial revascularization. PMID- 15346695 TI - [A comprehensive management system for heart failure improves clinical outcomes and reduces medical resource utilization]. AB - BACKGROUND: Hospital admissions for heart failure are common and readmission rates are high. Many admissions and readmissions may be avoidable, so that alternative strategies are needed to improve long-term management. METHODS: We conducted a randomized trial of the effect of a guideline-based intervention on rates of readmission within 90 days of hospital discharge and costs of care for patients who were hospitalized due to decompensated heart failure. The intervention consisted of comprehensive education of the patient and family, a prescribed diet and intensive application of guidelines' recommendations on pharmacological therapy. The intervention started before discharge and continued thereafter with follow-up visits for up to 3 months. Two hundred and nine guideline-managed patients were compared to 209 concurrent normally-discharged patients. RESULTS: Patients in the study group were more prescribed beta blockers, ACE-inhibitors, angiotensin receptor blockers, and spironolactone. Sixteen patients (8%) in the intervention group and 31 (15%) among controls were readmitted for DRG 127, within 3 months of discharge (Fisher's exact test, p < 0.01), while the 6-month mortality rate was similar between groups (9 and 11.5% respectively). Quality of life significantly improved from 5.6 +/- 1.0 to 6.1 +/- 1.9 (Mann-Whitney U-test, p < 0.05). The overall costs of care were lower for guideline-managed patients (110 vs 150 Euro per patient per month), due to the lower readmission rates. CONCLUSIONS: Our study showed that a guideline-based management program for patients with heart failure at discharge improves quality of life and reduces readmission for DRG 127 and total bed days, allowing relevant cost savings. PMID- 15346696 TI - [Repair of post-infarct left ventricular rupture with pericardial patch]. AB - The devolpment of a left ventricular rupture after myocardial infarction is not a rare complication. We report a case of 70-year-old male patient with clinical and instrumental signs of cardiac tamponade after left ventricular rupture. We repaired this lesion with pericardial patch and biological Glue without extracorporeal circulation and sutureless. PMID- 15346697 TI - [Heart failure in a subject treated with anthracyclines in childhood: myocardial dysfunction is not always the only reason of it]. AB - Anthracyclines, found to be efficacious in the treatment of a broad spectrum of pediatric malignancies, are cardiotoxic and may lead to heart failure even a long time after successful treatment of cancer. It is thought that subtle abnormalities can progress to the more permanent myocardial disease, resulting in cardiomyopathy which may progress to congestive heart failure. There are some precipitating factors leading to the sudden onset of cardiac symptoms such as increase in afterload or preload. We describe a young patient with congestive heart failure treated with doxorubicin (cumulative mean dose 420 mg/m2) in infancy because of pelvic sarcoma in whom the appearance of symptoms was related to pulmonary embolism. Four years before hospital admission, the patient presented echocardiographic abnormalities such as left ventricular fractional shortening and thickness reduction and he was treated with ACE-inhibitors. The myocardial ischemia, which is present in pulmonary embolism, probably worsened the left ventricular systolic function and caused congestive heart failure. PMID- 15346698 TI - [Task Force on the management of chest pain]. PMID- 15346699 TI - Dextromethorphan in pain control--an old drug with new clinical application. PMID- 15346700 TI - A local scavenging system to remove waste anesthetic gases during general anesthesia. AB - BACKGROUND: A local scavenging system was constructed and tested in both the operating room and the laboratory to remove the waste anesthetic gases so as to lower the exposure risk of the anesthetic personnel. METHODS: A local scavenging system was developed to suck away the waste anesthetic gases (e.g., N2O and sevoflurane) escaping from the mouth and nostrils of a patient. The local scavenging system used was composed of an inlet funnel (with a diameter of 20 cm), a flexible connecting tubing, a high efficiency particulate air (HEPA) filter and a vacuum pump. To help evaluate the performance of the local scavenging system, a tracer gas (SF6) of a fixed concentration (= 200 ppm) and flow rate (= 5 l/min) was introduced around the nostrils of the patient during anesthesia. The concentrations of the gases (SF6, N2O and SEV) drawn away by the scavenging system were then determined by an extractive Fourier transform infrared (FTIR) spectrometer and those spreading around the breathing zone of the anesthesiologist were obtained by the other FTIR. In the laboratory tests, the relationship between the scavenging efficiency and the inlet funnel position was obtained using the aforementioned SF6-FTIR techniques. RESULTS: With the application of this local scavenging system, during three surgical operations, the average personnel exposure concentrations of N2O and sevoflurane (SEV) as measured were 8.7 and 0.06 ppm, respectively. Both measured concentrations were lower than the TWA values recommended by the US-NIOSH for N2O (= 25 ppm) and SEV (= 2 ppm). Based on the tracer gas (SF6) results, it was found that the average scavenging efficiency was equal to 87%, which was lower than the laboratory testing results of 95%. The (scavenging) efficiency difference between the laboratory and on-site tests could be due to the movement and action of the anesthesiologist during anesthesia. To optimize the performance of the local scavenging device, the inlet (funnel) should be placed close to the breathing region (e.g., noses and mouth) of the patient in the front direction. CONCLUSIONS: The application of the local scavenging system was found to greatly reduce the concentrations of the waste anesthetic gases (e.g., N2O and SEV) to the levels lower than those recommended by the US-NIOSH. With this scavenging device, the exposure health risk of the anesthesiologists could be greatly reduced. PMID- 15346701 TI - A long-term analysis of the spinal somatosensory evoked potential of rats with sciatic constriction injury. AB - BACKGROUND: This study was undertaken to evaluate the long-term effect of a peripheral neuropathic pain model, chronic constriction injury (CCI) of the sciatic nerve, through analyses of electrophysiologic change, sciatic nerve function, and pain behavior. METHODS: CCI of the sciatic nerve was induced in twelve rats as described by Bennett and Xie. Three parameters were monitored: spinal somatosensory evoked potential (SSEP) elicited by supramaximal stimulation of the posterior tibial nerve and recorded from the thoracolumbar junctional space; thermal hyperalgesia assessed by measuring paw withdrawal latency (PWL); and sciatic function index (SFI). All the values of these parameters were obtained before the CCI procedure (day 1 as a preoperative baseline, and again on days 5, 12, 19, 26, 33, 47, 61, 75, and 89). SSEP was also measured 3 hr after the CCI operation. Data were also obtained from contralateral limbs. RESULTS: All rats with CCI developed thermal hyperalgesia on day 5, as indicated by a significant reduction in PWL in the CCI limbs and a deterioration of the SFI compared with baseline values. These effects persisted to day 89. In the electrophysiologic study, 3 hrs after the CCI operation, the amplitude significantly decreased and latency significantly increased in all SSEP recordings. The changes persisted and showed no further statistically significant deterioration or recovery. The data demonstrated that the major electrophysiologic change after a constriction injury was the loss of conduction ability across the injury site in the fast-conducting fibers, but in the slow conducting fibers the conduction ability was still preserved, which occurred immediately after the operation and persisted consistently for 89 days in the rats with behavior manifestations of neuropathic pain. CONCLUSIONS: Our findings suggest that SSEP is simple to obtain and sensitive in the acute phase of electrophysiological changes, but is limited for long-term evaluation after CCI. PMID- 15346702 TI - Effect of intravenous midazolam premedication on postoperative nausea and vomiting after cholecystectomy. AB - BACKGROUND: Several drugs and techniques have been used to reduce the incidence of postoperative nausea and vomiting (PONV). However PONV continues to be a common postoperative complication. Midazolam premedication in pediatric patients has been reported to reduce the incidence of PONV. In the present study the effect of intravenous midazolam premedication on the incidence and severity of PONV was investigated in a sample of adult patients undergoing anesthesia for cholecystectomy. METHODS: Eighty-two adult patients undergoing general anesthesia for cholecystectomy were randomly divided into two groups to receive either midazolam 75 microg/kg or a same volume of normal saline intravenously fifteen minutes prior to induction of anesthesia. Incidence and severity of PONV together with the total amount of administered metoclopramide during the first postoperative day were compared between two groups. RESULTS: Severity of nausea was significantly lightened in midazolam group during the first six hours after recovery period compared with placebo group (3.7 +/- 1.6 of a ten point visual analog scale vs. 4.9 +/- 2.2 in placebo group; P < 0.05). Mean number of vomiting episodes was significantly lower in midazolam group (0.4 +/- 0.7 vs. 1.1 +/- 1.4 in placebo group; P < 0.05). Midazolam group received a significantly less amount of metoclopramide during the first postoperative day (2.1 +/- 3.7 mg vs. 5.3 +/- 6.8 mg in placebo group; P < 0.05). CONCLUSIONS: The results of this study suggest the effectiveness of prophylactic intravenous midazolam premedication to reduce the incidence and severity of postoperative nausea and vomiting. Possible mechanisms for this effect of midazolam may be GABA receptor antagonism, inhibition of dopamine release, and anxiolytic effects. PMID- 15346703 TI - Predicting blood pressure change caused by rapid injection of propofol during anesthesia induction with a logistic regression model. AB - BACKGROUND: Propofol is a common intravenous agent for induction and maintenance of anesthesia. The advantage of propofol is rapid recovery of consciousness when the continuous infusion is stopped. Additionally, it has antiemetic effect of reducing postoperative nausea and vomiting. On the other hand, rapid infusion of propofol is painful and may cause hypotension. In this study, we aimed to develop a logistic regression model to accurately predict blood pressure change caused by rapid infusion of propofol. METHODS: Seventeen variables (including demographic data, past medical history, laboratory data, and blood pressure before induction) were assessed in 200 patients who received propofol for induction of anesthesia for routine surgery. A logistic regression model was derived using these values as independent variables to predict whether a patient would suffer a significant blood pressure change (> 30% decrease from baseline). Sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) were calculated to evaluate the performance of our prediction model. RESULTS: A cut-off value of 0.17 in the logistic regression model predicted decreased blood pressure with 90.0% sensitivity and, 67.3% specificity. The area under the receiver operating characteristic curve was 0.855. CONCLUSIONS: Our prediction model predicts propofol-induced hypotension with acceptable accuracy. Because of the straightforward mathematic formula used, our model can be integrated effortlessly into a hospital information system, providing a reliable and useful decision support for clinical anesthesia staff. PMID- 15346704 TI - Oral slow release dextromethorphan premedication provides a pethidine spare effect on posthemorrhoidectomy pain management. AB - BACKGROUND: In our previous study, we had demonstrated that intramuscular (i.m.) dextromethorphan (DM) could provide a preemptive analgesic effect and improve postoperative pain management. Regrow is a long-duration slow-release oral dextromethorphan available for clinical use with good patient compliance. The present study was designed to examine whether oral regrow may also offer the same preemptive analgesic effect as i.m. DM does in postoperative pain management. METHODS: Seventy-five patients, ASA status I and II, scheduled for hemorrhoidectomy were included and randomly assigned to the control and study groups. In the control group patients received placebo orally 8 h before surgical incision. In the study group, patients received regrow orally either 120 mg (R 120) or 240 mg (R-240) 8 h before skin incision. Pethidine (1 mg/kg, i.m.) was given for postoperative pain relief on demand. The time to first pethidine injection, total pethidine consumption, worst pain score, and pethidine-related side effects were recorded for two days. RESULTS: The times to first pethidine injection were 5.4 +/- 3.1, 6.5 +/- 3.5 and 12.7 +/- 5.7 h in the control, R-120 and R-240 groups, respectively. Total pethidine consumptions were 150 +/- 12, 132 +/- 11.8 and 82 +/- 12.5 mg in the control, R-120 and R-240 groups, respectively. The worst visual analog scale pain scores were respectively 7.2 +/- 0.4, 6.9 +/- 0.2 and 5.5 +/- 0.4 in the control, R-120 and R-240 groups during the 2-day observation. Five and three patients suffered pethidine-related side effects in the control and R-120 groups, respectively. CONCLUSIONS: This study revealed that premedication of oral regrow 240 mg provided a preemptive analgesic effect, thus reducing the severity of postoperative pain and pethidine requirement in post hemorrhoidectomy patients. PMID- 15346705 TI - Opioid tolerance: is there a dialogue between glutamate and beta-arrestin? AB - Opioids are widely used as analgesics in clinical pain management for decades. However, opioid tolerance is a serious problem which limits their usefulness. The mechanisms of opioid tolerance are complex which involve many regulatory factors. Glutamate is an important extracellular neurotransmitter which activates glutamate receptor and induces a series of signal transduction to regulate the development of opioid tolerance. Previous studies have indicated an involvement of NMDA receptors in the development of beta-opioid tolerance and associated abnormal pain sensitivity. Many NMDA receptor antagonists had been demonstrated to regulate morphine tolerance development. In the NMDA-mediated intracellular mechanisms of opioid tolerance, protein kinase C (PKC) modulates beta-opioid receptor activation. Besides, the opioid receptor desensitization involves phosphorylation of receptors and subsequent binds to beta-arrestin. In knockout mice, lacking beta-arrestin-2, desensitization of beta-opioid receptor did not occur after chronic morphine treatment, and these animals also failed to develop antinociceptive tolerance. Moreover, morphine tolerance can be completely reversed with a low dose of the classical PKC inhibitor chelerythrine in the beta arrestin-2 knock-out, but not wild-type mice. These findings indicate that, in the absence of beta-arrestin-2, contributions of PKC-dependent regulatory system would become apparent. In summary, PKC is regulated by NMDA receptors to affect the development of opioid tolerance, beta-arrestin-2 also influences PKC-induced opioid receptor desensitization. PKC may play an important role to coordinate these factors which regulate opioid tolerance. PMID- 15346706 TI - Anesthetic management of Boerhaave's syndrome. AB - Boerhaave's syndrome or spontaneous esophageal perforation is a life threatening condition which demands early diagnosis and urgent management. Although very selective patients can be treated nonoperatively, in most patients, better overall results can only be attained with early aggressive surgery. Clinical data and accounts of anesthetic management of this condition scarely appear in medical literature. Managing these patients for surgery is among the most challenging tasks facing the anesthesiologist because they may develop septic syndrome with shock. A rapid-sequence induction is mandatory, and the procedures that may aggravate the injury to esophagus should be avoided. Inotropic support and close attention to fluid balance may be required during operation. Because Boerhaave's syndrome is rare, we report here two cases to illustrate possible anesthetic implications of this disease. Both patients underwent thoracotomy to relieve empyema of mediastinum under general anesthesia. The first patient recovered completely after operation due to early diagnosis and treatment, but the second patient developed multiple organ failure and died after operation due to delayed diagnosis of esophageal rupture and severe sepsis. Because survival is directly related to the time to diagnosis and treatment, all clinicians need to be aware of this lethal disease. PMID- 15346707 TI - Fatal hemoptysis after suction through endotracheal tube in a patient with pneumoconiosis--a case report. AB - A 76-year-old man, with a history of pneumoconiosis, developed massive hemoptysis after suction through endotracheal tube. Emergent bronchial artery angiography revealed that rupture of an aneurysm was the suspected culprit and coil embolization was performed. However, the bleeding persisted and he eventually succumbed. Here we bring forward the clinical evaluation and management of massive hemoptysis for discussion. It is note worthy that in patients with chronic inflammatory lung diseases, such as pneumoconiosis, massive hemoptysis may develop in the act of trachcobronchial toilet with suction. PMID- 15346708 TI - High frequency jet ventilation in exploration of tracheostomy--a case report. AB - Exploration of tracheostomy is not uncommon, however, in case a patient with a facial reconstruction flap, trouble may arise because of difficulty in intubation. We hereby report a patient who had a fresh facial reconstruction flap after radical resection of buccal carcinoma, sustained bleeding around the tracheostomy and was scheduled for exploration of tracheostomy. Fiberoptic oral intubation was failed because of profuse secretion and edematous oral mucosa; high frequency jet ventilation (HFJV) was thus applied and made possible with a suction catheter put through the tracheostomy tube. Exploration was performed following withdrawal of the tracheostomy tube. The tracheostomy tube was reinserted under the guidance of the in-place suction tube after uneventful exploration. The probable concomitant complications and contraindications of using HFJV are discussed here. With thorough preparation and careful monitoring, and under the supervision of experienced physicians, transtracheal HFJV can substitute fiberoptic intubation for ventilation in case of exploration of tracheostomy. PMID- 15346709 TI - Postoperative transient ischemic attack complicated by CO2 retention and hypoxemia in a patient receiving meperidine injection for postoperative pain relief. AB - Transient ischemic attack (TIA) or stroke is not a rare complication, particularly in aged patients after surgery and anesthesia. If it is not properly managed, unmendable injury may inevitably occur. Patients who experience postoperative TIA or stroke usually have underlying diseases without resolution or good control. We bring forward here for discussion a fatality due to unappropriately managed TIA, which occurred in a 72-year-old male, within 24 hours after surgery. Besides making accurate diagnosis, selection of drugs for pain relief and appropriate overall management are key points of discussion. PMID- 15346710 TI - Successful detection and management of unexpected endotracheal tube kinking during neurosurgery--a case report. AB - Kinking of endotracheal tube (ETT) is not an infrequent problem during general anesthesia. However, failure to secure free airway following airway obstruction due to kinking of endotracheal tube or other tube problems may lead to lethality instead of lifesaving. We describe a case of unexpected kinking of ETT intraorally while craniotomy was underway. The kink might result from over bending of the softening tube in the oral cavity due to oral temperature and neck flexion. Early detection and immediate management of the ETT kinking may reduce the possibility of morbidity and mortality in anesthesia. It could be more difficult to carry out reintubation in such an awful situation when the operation was proceeding. In this case report we would like to highlight how to accurately and quickly diagnose and manage the ETT kinking intraorally while craniotomy was in process under anesthesia. Emphasis is also laid on the importance of properly positioning of the head and neck prior to any operation especially in craniotomy. Besides, the flexometallic (FM) or armoured endotracheal tubes may be good substitutes for the polyvinyl chloride (PVC) ones in preventing the ETT kinking during general anesthesia. PMID- 15346711 TI - You can't burnout unless you've been on fire! PMID- 15346712 TI - Developing a plan for pediatric spiritual care. AB - During life-changing events, people turn to spirituality for comfort, hope, and relief. This article raises nurses' awareness of and intent to provide spiritual care for children and families as part of overall quality care. Essential nursing knowledge for the development of a plan of care that includes the child's spirituality, religion, and culture, developmental stage, age-appropriate spiritual care activities, and the needs of the family are presented. PMID- 15346713 TI - Diethylstilbestrol exposure: it's still an issue. AB - Persons exposed to diethylstilbestrol have higher risks of certain health problems. Nurses have an important role in identifying women who received diethylstilbestrol while pregnant and the children of those pregnancies. Identification of these persons allows for risk analysis and counseling, and assists in appropriate referrals for further evaluation and treatment. The following information will help you facilitate this role. PMID- 15346714 TI - Know the benefits and risks of using common herbal therapies. AB - Although Americans are increasingly using herbal products for a variety of health conditions, they may not be aware of the demonstrated risks and benefits of these products. This study found public knowledge of 5 commonly used herbal products to be quite low, even among users of the investigated herbs. PMID- 15346715 TI - Healing ourselves, healing others: third in a series. AB - A review of the literature (Part 1 in this series) revealed the power and pitfalls of being female and being a healer. A qualitative study of medical surgical nurses (Part 2) gave a glimpse of the satisfaction, frustration, and confusion nurses experience as healers. In particular, participants did not engage in regular self-care behaviors and did not articulate ways to create healing environments that would support nurses as well as patients. In this third and final article, implications and recommendations beyond the study will be shared. Education, clinical practice, leadership, and research are aspects of professional nursing that may benefit from these interpretations. PMID- 15346716 TI - Remember me: mask making with chronically and terminally ill children. AB - Mask making is one of many alternative methods nurses can integrate into the care of chronically and terminally ill children. Healthcare systems abound with materials that may be used in this endeavor. This article explores how mask making offers children an opportunity to gain some control over their environment and thus heal themselves, even in the worst of circumstances. PMID- 15346717 TI - Acupuncture usage shows relief of chronic symptoms. PMID- 15346718 TI - Copper treatment alters the barrier functions of human intestinal Caco-2 cells: involving tight junctions and P-glycoprotein. AB - This study investigated the effects of copper on paracellular permeability and P glycoprotein (P-gp) in Caco-2 cells. Apical treatment with 100-300 microM CuSO4 in Hanks' balanced salt solution (HBSS, up to 3 hours) induced a time- and concentration-dependent increase in permeability of Caco-2 cell monolayers monitored by transepithelial electrical resistance (TEER). Copper treatment also induced a concentration-dependent reduction of F-actin stain, but not of tight junctional protein ZO-1. In addition, without any adverse effects on TEER, apical treatment with 300 microM CuSO4 in complete medium (for 24 hours) could reduce basolateral-to-apical transport, and increase apical-to-basolateral transport of rhodamine-123 (Rho-123) and accumulation of Rho-123 in Caco-2 cells. Treatment with 10-100 microM CuSO4 in HBSS (up to 3 hours) also induced a time- and concentration-dependent increase in accumulation of Rho-123 in Caco-2 cells. The results indicated that copper treatment increased the paracellular permeability probably by perturbing F-actin skeleton, and inhibited P-gp, thus altering the barrier functions of Caco-2 cells. PMID- 15346719 TI - Aniline: early indicators of toxicity in male rats and their relevance to spleen carcinogenicity. AB - Early indicators of aniline hydrochloride (AH) toxicity were investigated in male Fisher 344 rats for 1 or 4 weeks at dietary dose levels of 10, 30 or 100 mg/kg body weight (bw)/day (actual intake at least 6, 17 and 57 mg/kg). The doses were based on earlier studies that had shown spleen toxicity and carcinogenicity in male rats at 100 mg/kg/day but not at 10 mg/kg/day. In the present study a dose related formation of haemoglobin adducts and Heinz bodies was found from 10 and 30 mg/kg bw/ day, respectively, onwards. Signs of anaemia (decreased red blood cell counts and increased reticulocytes) were recorded from 30 mg/kg onwards. At 100 mg/kg, an overt haemolytic anaemia was associated with increases in serum transferrin concentration and total iron binding capacity in the blood reflecting major perturbations in iron metabolism. At this dose there was an increase in peripheral neutrophil leucocytosis in the blood, indicating an inflammatory process in the spleen. Histopathologic evaluation showed a focal perisplenitis and haemosiderin deposition in sinusoidal Kupffer cells of the liver at 100 mg/kg. These results corroborate the contention that carcinogenic doses of aniline cause early effects on haematological parameters, inflammatory reaction in the spleen and perturbations in iron metabolism as a result of haemolytic anaemia. Accordingly, the carcinogenicity of aniline may be linked to definable threshold-related processes. PMID- 15346720 TI - The protective effect of vitamin C, vitamin E and selenium combination therapy on ethanol-induced duodenal mucosal injury. AB - In this study, the effect of a combination of vitamin C, vitamin E and selenium on ethanol-induced duodenal mucosal damage in rats was investigated morphologically and biochemically. The duodenal mucosal injury was produced by oral administration of 1 mL of absolute ethanol to each rat. Animals received vitamin C (250 mg/ kg), vitamin E (250 mg/kg) and selenium (0.5 mg/kg) for 3 days and absolute ethanol 1 hour after last antioxidant administration and were sacrificed 1 hour after absolute ethanol. Extreme degeneration in intestinal mucosa of rats given ethanol was observed morphologically. In addition, an increase in neuronal nitric oxide synthase immunoreactive areas was observed in the rats of the group given ethanol. On the other hand, a normal morphological appearance and a decrease in neuronal nitric oxide synthase immunoreactive areas were detected in the rats given ethanol+vitamin C+vitamin E+ selenium. In the group to which ethanol was administered, an increase in serum cholesterol and a decrease in serum albumin levels were determined. On the other hand, in the group to which ethanol+vitamin C+vitamin E+selenium were administered, serum cholesterol value decreased, and the serum albumin level increased. As a result, we can say that the combination of vitamin C, vitamin E and selenium has a protective effect on ethanol-induced duodenal mucosal injury. PMID- 15346721 TI - Male reproductive toxicity of sodium arsenite in mice. AB - The effect of chronic oral exposure to arsenic on male mouse testicular and accessory sex organ weights, sperm parameters and testicular marker enzymes was studied. In addition, the distribution of arsenic in reproductive organs was measured using atomic absorption spectrophotometry. Sodium arsenite administered to mice (Mus musculus) via drinking water at a dose of 53.39 micromol/L (4 ppm As) for 365 days caused a decrease in the absolute and relative testicular weight. However, epididymal and accessory sex organ weight was similar to control. The activities of marker testicular enzymes such as sorbitol dehydrogenase, acid phosphatase and 17beta-hydroxy-steroid dehydrogenase (17beta HSD) were significantly decreased, but those of lactate dehydrogenase and gamma glutamyl transpeptidase (gamma-GT) were significantly increased. A decrease in sperm count and sperm motility, along with an increase in abnormal sperm, was observed in arsenite-exposed mice. A significant accumulation of arsenic in testes, epididymis, seminal vesicle and prostate gland was observed in treated animals. Thus long term exposure (365 days) at the dose level of 53.39 micromol/L sodium arsenite (4 ppm As), to which human beings are likely to be exposed via drinking water, may cause testicular and spermatotoxic effect. PMID- 15346722 TI - Statistical evaluation of an acute dermal toxicity test using the dermal fixed dose procedure. AB - The conventional method for the assessment of acute dermal toxicity (OECD Test Guideline 402, 1987) uses death of animals as an endpoint to identify the median lethal dose (LD50). A new OECD Testing Guideline called the dermal fixed dose procedure (dermal FDP) is being prepared to provide an alternative to Test Guideline 402. In contrast to Test Guideline 402, the dermal FDP does not provide a point estimate of the LD50, but aims to identify that dose of the substance under investigation that causes clear signs of nonlethal toxicity. This is then used to assign classification according to the new Globally Harmonised System of Classification and Labelling scheme (GHS). The dermal FDP has been validated using statistical modelling rather than by in vivo testing. The statistical modelling approach enables calculation of the probability of each GHS classification and the expected numbers of deaths and animals used in the test for imaginary substances with a range of LD50 values and dose-response curve slopes. This paper describes the dermal FDP and reports the results from the statistical evaluation. It is shown that the procedure will be completed with considerably less death and suffering than guideline 402, and will classify substances either in the same or a more stringent GHS class than that assigned on the basis of the LD50 value. PMID- 15346723 TI - Genoprotective role of vitamin E and selenium in rabbits anaesthetized with sevoflurane. AB - In this study, genotoxic effects of repeated sevoflurane anaesthesia were investigated in rabbits with or without antioxidant supplementation. Twenty-one New Zealand male rabbits were included in the study and randomized into three groups as: placebo treated (Group I), vitamin E supplemented (Group II) and selenium supplemented (Group III). Vitamin E and selenium were given intraperitoneally for 15 days before anaesthesia treatment. Anaesthesia was administered using 3% sevoflurane in 4 L/min oxygen for a 3-hour period and continued for 3 days. Blood samples were collected before anaesthesia (Sample 1), after the first, second and third days of sevoflurane administration (Sample 2, Sample 3 and Sample 4 respectively) and the last samples were taken 5 days after the last sevoflurane administration (Sample 5). Genotoxic damage was examined using the comet assay. The degree of damage is assessed by grading the cells into three categories of no migration (NM), low migration (LM) and high migration (HM) depending on the fraction of DNA pulled out into the tail under the influence of the electric field. The number of comets in each sample was calculated (1 x number of comets in category NM+2 x number of comets in category LM+3 x number of comets in category HM) and expressed as the total comet score (TCS), which summarizes the damage frequencies. In Group I, a significant increase in the mean TCSs was observed for Samples 3 and 4 as compared with Sample 1. However, there were no significant differences between Samples 1, 2 and 5. The mean TCS of Sample 4 was significantly higher than Sample 1, 2 and 3 in Group II. Group III demonstrated no significant mean TCSs for any experimental conditions. Statistical differences were also observed between the groups with significant P values. This experimental study points out the presence of DNA damage with repeated sevoflurane anaesthesia and the genoprotective role of antioxidant supplementation on DNA damage in mononuclear leukocytes of rabbits by highly sensitive comet assay. PMID- 15346724 TI - 401(k) plan asset allocation, account balances, and loan activity in 2003. PMID- 15346725 TI - Infection after cholecystectomy, hysterectomy or appendectomy. AB - OBJECTIVES: This article uses patient-linked data to focus on hospitalization with post-operative infection following cholecystectomy, hysterectomy or appendectomy. The average number of hospital days and the costs of readmission are also estimated. DATA SOURCE: Data for surgeries in fiscal years 1997/98, 1998/99 and 1999/00 are from the Health Person-Oriented Information Database. ANALYTICAL TECHNIQUES: Bivariate tabulations were used to estimate the percentage of patients hospitalized with post-operative infection after cholecystectomy, hysterectomy or appendectomy between 1997/98 and 1999/00. Logistic regression was used to explore associations between infection and patient characteristics, readmission, and peri-operative mortality, while controlling for surgical characteristics. MAIN RESULTS: Hospitalization with post-operative infection was relatively rare, occurring in 1.4% of cholecystectomy, 2.0% of hysterectomy, and 3.8% of appendectomy patients. The associated costs of readmission for post operative infection for the three surgeries were estimated at 5.4 to 6.3 million dollars annually. Old age, being male, surgical complexity and approach, and diabetes were associated with hospitalization involving a post-operative infection. PMID- 15346726 TI - Inflammatory bowel disease--hospitalization. AB - OBJECTIVES: This analysis examines trends in hospitalization for Crohn's disease and ulcerative colitis, the two main forms of inflammatory bowel disease (IBD). DATA SOURCES: Data are from the Hospital Morbidity Database for 1983/84 to 2000/01, and from the Health Person-Oriented Information Database for 1994/95 to 2000/01. ANALYTICAL TECHNIQUES: Sex- and age-specific rates were calculated for separations attributed to Crohn's disease and ulcerative colitis. Rates and hospital days were also calculated for hospitalizations in which IBD was among the first five diagnostic codes on a patient's discharge abstract. The frequency of rehospitalization was examined. MAIN RESULTS: From the early 1980s to the mid 1990s, annual rates of hospitalization for Crohn's disease and ulcerative colitis rose slightly, but have since levelled off. Hospitalization rates for both conditions are highest among people in their twenties. The average length of stay for patients with either disease fell from about 2 weeks in 1983/84 to 9 or 10 days in 2000/01. More than a quarter of patients hospitalized for Crohn's disease and over 20% of those with ulcerative colitis were readmitted within the same year. PMID- 15346727 TI - Use of cannabis and other illicit drugs. PMID- 15346728 TI - Creutzfeldt-Jakob disease. PMID- 15346729 TI - Pregnancy and smoking. PMID- 15346730 TI - Cobalt-based alloys for orthopaedic applications studied by electrochemical and XPS analysis. AB - The composition of the passive layers formed by electrochemical oxidation at different passivation potentials on Co-Cr-Mo and Co-Ni-Cr-Mo alloys in simulated physiological solution (SPS), with and without the complexing agent EDTA, was studied by X-ray photoelectron spectroscopy. Composition as a function of depth, cationic fraction and thickness of the passive film was determined. Chromium oxide is shown to be the major constituent of the passive layer on both Co-Cr-Mo and Co-Ni-Cr-Mo alloys. The minor constituents of the passive layers, Co- and Mo oxide in the case of Co-Cr-Mo alloy and Ni-, Co- and Mo-oxides in the case of Co Ni-Cr-Mo alloy, are also located in the outer part of the layer. EDTA affects the formation of the passive layer on each alloy. The content of Co-, Ni- and Mo oxide in the passive layer is lower in the presence of EDTA, thus indicating increased solubility associated with higher stability constants for complexes of metal cations with EDTA. PMID- 15346731 TI - Mechanical failure of human fetal membrane tissues. AB - Mechanical integrity of the chorioamnion membrane, and the component chorion and amnion layers, was assessed with biaxial puncture testing. Fetal membranes were obtained from term placentas following labored natural delivery or scheduled cesarean section. Preterm specimens were obtained from deliveries prior to 37 weeks gestation. Dividing and peripheral membranes were obtained from multiple gestation pregnancies. Specimens were gripped between parallel plates with circular openings and loaded with an instrumented, hand-held blunt probe until rupture occurred. Peak force was recorded and rupture sites were examined. Defects in multi-layered membranes differed in both size and shape in the individual layers. Compared with chorion and whole chorioamnion, amnion was more mechanically sensitive to different obstetrical conditions. Amnion varied in response at different physical locations within the same patient. Membrane and component puncture force data were used to calculate biaxial failure strength. Membrane stresses arising from amniotic fluid pressure were computed as a function of gestational age, and compared to membrane strength to examine the criterion for membrane failure in vivo. Possible mechanical conditions for preterm membrane rupture were examined. PMID- 15346732 TI - Physical characteristics of polymer complexes in suspension obtained from cellulosic latexes with ondansetron. AB - Ondansetron is a carbazol with antiemetic properties. It is used primarily to control nausea and vomiting caused by cytotoxic chemotherapy and radiotherapy, as well as in postoperative vomiting in gynecological surgery. Ondansetron has a half-life of approximately 4 h, hence it is a matter of great interest to determine the ideal conditions for the formation of a drug-polymer complex in order to prolong the duration of the therapeutic action. A stability study of the active drug was first carried out on each of the polymers (Aquateric and Aquacoat). The adsorption of ondansetron on the lattices was determined with respect to time, pH and concentration. The results obtained suggest that both polymers are suitable as drug carriers for the controlled-release formulations obtained. We conclude that an acid pH is evidently fundamental in the adsorption process of this drug in the latexes. Moreover, the Aquateric latex would seem to be the best-suited polymer to use as a vehicle for drug delivery. PMID- 15346733 TI - Mechanical and biological properties of hydroxyapatite reinforced with 40 vol. % titanium particles for use as hard tissue replacement. AB - Hydroxyapatite (HA)-based composite reinforced with 40 vol. % Ti particles was fabricated by the optimal technical condition of hot pressing technique. The mechanical and biological properties of the composite were studied by mechanical and in vivo methods. The experimental results show that HA and Ti phases are the predominant phases of the composite with partially decomposition of HA phase into alpha-Ca3(PO4)2 and Ca4O(PO4)2. Comparing with HA-20 vol. % Ti composite manufactured under the same conditions, HA-40 vol. % Ti composite with similar elastic modulus (79.3 GPa) and Vicker's hardness (2.94GPa) has a higher bending strength (92.1 MPa). Moreover, fracture toughness of HA-40 vol. % Ti composite with crack bridging as the chief toughening mechanisms can reach 2.692 MPa m(-1) , which can meet the basic toughness demand of the replaced hard tissues for heavy load-bearing applications. Work of fracture of HA-40 vol. % Ti composite is 91.2J m2, which is 22.9 times that of pure HA ceramic and even 2.4 times that of Al2O3 bioceramic. The results of in vivo studies show HA-40 vol. % Ti composite has excellent biocompatibility and could integrate with bone. In the early stage after the implantation of the samples, the osteointegration ability of the composite is better than that of pure titanium. PMID- 15346734 TI - Silk fibroin modified porous poly(epsilon-caprolactone) scaffold for human fibroblast culture in vitro. AB - In order to develop scaffolds with improved biocompatibility for cell culture, hybrid scaffolds were fabricated by modifying poly(epsilon-caprolactone) (PCL) with silk fibroin (SF) in a porous structure. Scanning electronic microscopy revealed that the morphology of the PCL-SF hybrid scaffold was affected by the concentration of the SF solution. Availability of SF on the surface and the conformational transition induced by methanol treatment were proved by attenuated total reflection Fourier transformed infrared spectroscopy (ATR-FTIR), and wettability of the hybrid scaffold was greatly improved. To evaluate scaffold biocompatibility, human fibroblasts were cultured on the hybrid scaffold with the unmodified PCL scaffold as control. An MTT assay indicated that although fewer cells were initially held on the hybrid scaffold after one day of culture, comparable cell numbers were achieved after four days and significantly more cells proliferated on the hybrid after seven days. The cell morphology also indicated that the PCL-SF hybrid scaffold was favorable for cell culture. This study suggests that surface modification with SF would be an effective way to improve the biocompatibility of PCL, facilitating its application in practical tissue engineering. PMID- 15346735 TI - Synthesis and characterisation of poly(2-hydroxyethyl methacrylate) polyelectrolyte complexes. AB - Copolymerisation of charged and neutral monomers is a well-known methodology to introduce charged moieties in a polymeric chain to obtain polyelectrolytes. New polyelectrolyte complexes have been synthesised by radical copolymerisation of neutral methacrylic monomer 2-hydroxyethyl methacrylate (HEMA) with cationic 2 methacryloyloxyethyltrimethyl ammonium chloride and anionic 2-acrylamido-2 methylpropane-sulphonic acid monomers in 10:1:1 and 10:1:2 stechiometric ratio. Chemical structure of the synthesised terpolymers was confirmed by FT-IR spectroscopy, moreover, X-ray photoelectron spectroscopy showed the presence of a cationic charge excess on the 10:1:2 terpolymer surface with respect to 10:1:1 terpolymer. Swelling studies for 10:1:2 terpolymers showed a high water content in the swollen state and a "smart behaviour" upon changes in external stimuli such as pH, while, 10:1:1 terpolymer presented the behaviour of a neutral polymer. Mechanical and differential scanning calorimetry analysis confirmed that terpolymer networks were stabilised by ionic co-operative interactions. Infact, the inclusion of oppositely ionic charges in the polymeric network of p(HEMA) represent a way to achieve higher elastic modulus as they stabilise the terpolymer networks. Cytotoxicity and cytocompatibility studies demonstrated that all materials were not toxic, moreover, the presence of a cationic charge excess on 10:1:2 terpolymer surface was able to promote fibroblast adhesion. PMID- 15346736 TI - Effects of Ce on the short-term biocompatibility of Ti-Fe-Mo-Mn-Nb-Zr alloy for dental materials. AB - Effects of Ce on the short-term biocompatibility of Ti-Fe-Mo-Mn-Nb-Zr alloy designed for implant materials were studied by acute toxicity test, hemolytic test, and MTT assay. The elements and their concentration in surface films and extraction media of Ti alloys were investigated with XPS and ICP, respectively. The primary compositions of the surface films of Ti alloys with 0.3% Ce and without Ce were TiO2 and Nb2O5. There were 0.2 mg/l Fe and 0.16 mg/l Mn in the extraction medium of Ti alloy without Ce, while 0.27 mg/l Fe and 0.87 mg/l Mn in the extraction medium of Ti alloy with 0.3% Ce. The concentrations of Fe and Mn in the medium were too low to have any significant effects on human health. There was no sign of cytotoxicity in these tests. The cytotoxicity levels of Ti alloys without Ce and with 0.3% Ce were graded 0 and 1, respectively. The hemolytic degrees of Ti alloys without Ce and with 0.3% Ce were 0.558% and 0.67%, respectively. The cells being incubated in the extraction medium were normal. These phenomena indicated that Ce was innocuous within the concentration range of this study. In addition, the hemolytic ratio and toxicity level of Ti alloy with 0.3% Ce were a little higher than that of Ti alloy without Ce. This meant that Ce would slightly increase the toxicity of Ti alloy. PMID- 15346737 TI - Osteoblast response and calcium deposition on phospholipid modified surfaces. AB - In this study, the effect of calcium phosphate complexed phospholipid (Ca-PL-PO4) coatings on solid surfaces on the in vitro calcium (Ca) deposition and on the osteoblast responses was evaluated. Commercially available phospholipids were converted to their Ca-PL-PO4, and were coated on glass Petri dishes. The coated dishes were immersed in the simulated body fluid for up to 14 days under sterilized conditions at 37 degrees C, and the amount of calcium (Ca) deposited was quantified. Similarly, by measuring the alkaline phosphatase specific activity, the differentiation of osteoblast precursor cells were evaluated after seeding the cells on Ca-PL-PO4 coated cell culture plastics. It was observed that all Ca-PL-PO4 enhanced Ca deposition on coated surfaces. The, polar head group of phospholipids in coated surfaces was observed to have an influence on the Ca deposition as well as the osteoblast differentiation. Among the phospholipids evaluated, phosphatidylserine (Ca-PS-PO4) exhibited the strongest calcium deposition and more enhanced alkaline phosphatase specific activity. It was therefore concluded from this study that Ca-PS-PO4 surface modification may be an alternative method for enhancing bone-implant interactions. PMID- 15346738 TI - Screening biomaterials with a new in vitro method for potential calcification: porcine aortic valves and bovine pericardium. AB - Calcification is still a major cause of failure of implantable biomaterials. A fast and reliable in vitro model could contribute to the study of its mechanisms and to testing different anticalcification techniques. In this work, we attempted to investigate the potential calcification of biomaterials using an in vitro model. We purposed to test the ability of this model to screening possible anticalcification efficacy of different biomaterials. Porcine heart valve (PAV) and bovine pericardial (BP) tissues, fixed with glutaraldehyde were immersed into biological mimicking solution, where the pH and the initial concentrations of calcium and phosphoric ions were kept stable by the addition of precipitated ions during calcification. Kinetics of calcification was continuously monitored. The evaluation of biomaterials was carried out by comparing the kinetic rates of formation of calcific deposits. After 24 h, the calcific deposits on PAVs were found to be developed at significant higher rates (ranged from 0.81 x 10(-4)-2.18 x 10(-4)mol/min m2) than on BP (0.19 x 10(-4)-0.52 x 10(-4)mol/min m2) (one-way ANOVA, p < 0.05) depending on the experimental conditions (supersaturation of the solution). Parallel tests for similar biomaterials implanted subcutaneously in animal (rat) model showed after 49 days that significant higher amounts of total minerals deposited on PAV (236.73+/-139.12, 9 animals mg minerals/g dry net tissue) (mean+/-standard deviation) compared with that formed on BP (104.36+/ 79.21, #9 mg minerals/g dry net tissue) (ANOVA, p < 0.05). There is evidence that in vitro calcification was correlated well with that of animal model and clinical data. PMID- 15346739 TI - The effect of hot pressing on the physical properties of glass reinforced hydroxyapatite. AB - Hydroxyapatite (HA), being of physiological importance, can be developed synthetically for implant application. A number of avenues have been explored in order to improve the physical and biological properties of a variety of hydroxyapatite composites. However, the fact remains, hydroxyapatite lacks the mechanical properties needed to sustain high loads. This study investigates the advantages of hot pressing on the physical properties of HA and glass reinforced HA (GR-HA). The results show a significant enhancement in the mechanical properties of GR-HA composites compared to HA e.g. flexural bending strength values were given at 91.75 and 88.87 M Nm(-2) for GR-HA (CP15F) and GR-HA (CP20F) respectively, compared to 78.9 M Nm(-2) for HA. The results for other properties such as elastic modulus, fracture toughness, Vicker's hardness, density and porosity also demonstrate the benefit of adding phosphate based glasses as a sintering aid. This is supported by XRD analysis, highlighting the presence of a secondary phase (beta-TCP) in GR-HA systems and the positive effect it has on the physical properties. It must be brought to attention that densification of hot pressed HA and GR-HA composites is reached at a lower temperature compared to a previous study on the same materials that have undergone pressureless sintering. PMID- 15346740 TI - Differences between the wear couples metal-on-polyethylene and ceramic-on-ceramic in the stability against dislocation of total hip replacement. AB - After total hip replacement an insufficient range of motion (ROM) can lead to contact between femoral neck and rim of the cup (= impingement) causing dislocation and consecutive material failure. The purpose of this study was to analyse the influence of different wear couples on the ROM and stability against dislocation. By means of a special testing device the ROM until impingement, the ROM until dislocation as well as the resisting moment against levering the head out of the cup were experimentally determined. Various total hip systems with cup inserts made of ceramic and polyethylene were comparatively examined in different implant positions. Maximum resisting moment as well as the ROM until impingement and dislocation were clearly influenced by the implant position. Furthermore, the stability against dislocation was affected by design parameters, whereas in the case of appropriate implant position differing wear couples (metal-on polyethylene vs. ceramic-on-ceramic) had a minor impact. However, as shown by tests under lubricant conditions, ceramic-on-ceramic couples provided less dislocation stability in unfavourable implant position in comparison to metal-on polyethylene. Therefore, ceramic-on-ceramic couples should only be applied in the case of optimised implant orientation preventing impingement and dislocation with subsequent material failure like chipping off or breakage. PMID- 15346741 TI - Tissue response to the implantation of biodegradable polyhydroxyalkanoate sutures. AB - Polyhydroxyalkanoate (PHA) sutures were implanted to test animals intramuscularly, and tissue reaction was investigated and compared with the reaction to silk and catgut. Tested monofilament sutures made of PHAs of two types--polyhydroxybutyrate (PHB) and a copolymer of hydroxybutyrate and hydroxyvalerate (PHV)--featured the strength necessary for the healing of muscle fascial wounds. The reaction of tissues to polymeric implants was similar to their reaction to silk and was less pronounced than the reaction to catgut; it was expressed in a transient post-traumatic inflammation (up to four weeks) and the formation of a fibrous capsule less than 200 microm thick, which became as thin as 40-60 microm after 16 weeks, in the course of reverse development. Macrophages and foreign-body giant cells with a high activity of acid phosphatase were actively involved in this process. PHB and PHB/PHV sutures implanted intramuscularly for an extended period (up to one year) did not cause any acute vascular reaction at the site of implantation or any adverse events, such as suppurative inflammation, necrosis, calcification of the fibrous capsule or malignant tumor formation. No statistically significant differences were revealed in the tissue response to polymer sutures of the two types. Capsules around silk and catgut sutures did not become significantly thinner. PMID- 15346742 TI - In vivo characterisation of a novel bioresorbable poly(lactide-co-glycolide) tubular foam scaffold for tissue engineering applications. AB - Polylactide-co-glycolide (PLGA) foams of tubular shape were assessed for their use as soft-tissue engineering scaffolds in vitro and in vivo. Porous membranes were fabricated by a thermally induced phase separation process of PLGA solutions in dimethylcarbonate. The parameters investigated were the PLGA concentration and the casting volume of solution. Membranes produced from 5 wt/v % polymer solutions and a 6 ml casting volume of polymer solution were selected for fabricating tubes of 3 mm diameter, 20 mm length and a nominal wall thickness of 1.5 mm. Scanning electron microscopy revealed that the structure of the tubular foams consisted of radially oriented and highly interconnected pores with a large size distribution (50-300 microm). Selected tubes were implanted subcutaneously into adult male Lewis rats. Although the lumen of the tubes collapsed within one week of implantation, histological examination of the implanted scaffolds revealed that the foam tubes were well tolerated. Cellular infiltration into the foams, consisting mainly of fibrovascular tissue, was evident after two weeks and complete within eight weeks of implantation. The polymer was still evident in the scaffolds after eight weeks of implantation. The results from this study demonstrate that the PLGA tubular foams may be useful as soft-tissue engineering scaffolds with modification holding promise for the regeneration of tissues requiring a tubular shape scaffold such as intestine. PMID- 15346743 TI - TEM and molecular simulation studies on the hydroxylapatite structure with Si and Mg impurities. AB - Transmission electron microscopy (TEM) and molecular simulation studies of traces of chemical elements such as Mg, Si, and OH in the hydroxylapatite (CaHAP) crystal structure obtained from the sand dollar were carried out. Two different types of CaHAP crystal morphologies in the samples synthesized by the hydrothermal method used were observed. Reflections with regular intensity in the experimental electron diffraction patterns obtained from these morphologies were observed. However, when these results were compared with a simulated electron diffraction pattern (which was obtained using the crystalline structure proposed for the hydroxylapatite) some forbidden reflections in the experimental pattern were observed. Then, in order to reproduce the experimental patterns Si and Mg atoms in the crystalline lattice were introduced. These elements in the elemental chemical analysis carried out by X-ray energy dispersive spectroscopy (EDS) in the typical CaHAP morphologies were detected. The positions of these atoms in the asymmetric unit were obtained using molecular simulation and during the relaxation process, the structure did not show changes in the lattice parameters. Subsequently, the crystalline structure was reproduced and matched the electron diffraction patterns simulated resulting in the experimental electron diffraction pattern. Experimental and simulated X-ray diffraction spectra were also matched. PMID- 15346744 TI - Noting a most notable retirement. PMID- 15346745 TI - Comparison of an allograft in an experimental putty carrier and a bovine-derived xenograft used in ridge preservation: a clinical and histologic study in humans. AB - PURPOSE: The aim of this randomized, controlled, blinded clinical study was to compare ridge dimensions and histologic characteristics of ridges preserved with 2 different graft materials. MATERIALS AND METHODS: Twenty-four subjects, each requiring a nonmolar extraction and delayed implant placement, were randomly selected to receive ridge preservation treatment with either an allograft in an experimental putty carrier plus a calcium sulfate barrier (PUT) or a bovine derived xenograft (BDX) plus a collagen membrane. Horizontal and vertical ridge dimensions were determined using a digital caliper and a template. At 4 months postextraction, a trephine core was obtained for histologic analysis. RESULTS: The average ridge width decreased by 0.50 mm for both groups (P < .05). The midbuccal vertical change for the PUT group was a loss of 0.3+/-0.7 mm versus a gain of 0.7+/-1.2 mm for the BDX group, a difference of 1.0 mm (P > .05). Histologic analysis revealed vital bone in the PUT group of about 61%+/-9% versus 26%+/-20% for the BDX group (P < .05). DISCUSSION: Greater vital bone fill in the PUT group may be attributable to earlier and greater vascular invasion of the carrier material. The putty material was characterized by ease of handling, simple placement, and enhanced graft particle containment. CONCLUSIONS: Allograft mixed with an experimental putty carrier produced significantly more vital bone fill than did the use of a xenograft with no carrier material. Ridge width and height dimensions were similarly preserved with both graft materials. PMID- 15346746 TI - Long-term bone response to titanium implants coated with thin radiofrequent magnetron-sputtered hydroxyapatite in rabbits. AB - PURPOSE: The present study was designed to investigate the long-term bone response around machined screw-type uncoated and calcium phosphate (CaP) -coated commercially pure titanium implants. MATERIALS AND METHODS: Using a magnetron sputtering technique, implants with a CaP coating similar in composition and CaP ratio to hydroxyapatite were produced. Heat treatment was subsequently used to increase the crystallinity of the coatings. Four types of coatings (0.1 and 2.0 microm amorphous and 0.1 and 2.0 microm crystalline) were manufactured; uncoated implants served as a control. Three hundred twenty implants (64 of each type) were randomly placed in the tibial cortical and trabecular femoral bones of 40 rabbits. The rabbits were sacrificed 9 months after implant placement. RESULTS: Histomorphometric evaluation carried out on ground sections revealed that the crystalline CaP coatings achieved the highest bone-implant contact in both tibiae and femora compared with amorphous CaP-coated and uncoated titanium. DISCUSSION: The present study suggests that submicron crystalline hydroxyapatite coating adds bioactive properties to titanium oral implants. CONCLUSION: An ultra-thin, 0.1 microm crystalline CaP coating can elicit and maintain an improved long-term bone response compared to amorphous coated or uncoated Ti implants, without any adverse tissue reactions. PMID- 15346747 TI - Direct assessment of profilometric roughness variability from typical implant surface types. AB - PURPOSE: Protocols for quantifying the surface roughness of implants are varied and dependent upon the roughness parameter produced by the particular measurement device. The objective of this study was to examine the accuracy and precision of typical roughness characterization instruments used in the dental implant industry. MATERIALS AND METHODS: The average roughness (Ra) was measured using 2 common surface characterization instruments: an interferometer and a stylus profilometer. Titanium disks were prepared to represent 4 typical dental implant surfaces: machined, acid-etched, hydroxyapatite-coated, and titanium plasma sprayed. Repeated measurements from multiple sites on each surface were undertaken to establish statistical inferences. Qualitative images of the surfaces were also acquired using a laser scanning confocal microscope. After surface measurements were conducted, the disks were diametrically cut and cross sectional profiles were examined using a scanning electron microscope (SEM) as a comparative measure of surface topography. An analysis of variance was applied to isolate the effects of the measurement site, measurement sequence, surface treatment, and instrument type on Ra values. RESULTS: The results indicated that surface treatment (P = .0001) and instrument (P = .0001) strongly influenced Ra data. By design, measurement site (diametrical: P = .9859; area: P = .9824) and measurement sequence (P = .9990) did not influence roughness. In the assessment of individual instrument accuracy, the interferometer was the most accurate in predicting SEM-based roughness (P = .6688) compared with the stylus (P = .0839). As a measure of aggregate precision over all measurements, the most repeatable instrument was the stylus (coefficient of variation [CV] = 0.108), followed by the interferometer (CV = 0.125) and SEM (CV = 0.273). DISCUSSION: These results indicate dependencies in accuracy and precision related to the surface characterization technique. CONCLUSION: Instrument variability may obscure functional correlations between implant surface topography and osseointegration. PMID- 15346748 TI - Accuracy of 2 impression techniques for ITI implants. AB - PURPOSE: The aim of this study was to compare the accuracy of casts produced by 2 impression techniques and elastomeric impression materials commonly used for the fabrication of implant-supported fixed prostheses. MATERIALS AND METHODS: A master model with 4 synOcta ITI implants placed unilaterally in place of the mandibular right central incisor, canine, first premolar, and first molar was constructed. Implant-level impressions were made by direct and indirect techniques. In the direct technique, synOcta impression caps with integral guide screws were used to transfer the implants using a custom-made acrylic resin tray and a polyether impression material (the PE direct technique). In the indirect technique, synOcta plastic positioning cylinders with impression caps were used to transfer the implants with either a custom-made acrylic resin tray and polyether impression material (the PE indirect technique) or with a stock tray with a vinylpolysiloxane impression material (the VPS indirect technique). After impression making, all casts (n = 21) were poured in type IV dental stone. Linear changes in -x or -y direction and numeric and descriptive angular changes between the implants were quantified using a coordinate measuring machine. RESULTS: Seven of 12 distance measurements (6 for -x direction, 6 for -y direction) showed differences between groups (P < .05). Of these, 5 were associated with the PE direct versus PE indirect and PE indirect versus VPS indirect, and 3 were associated with PE direct versus VPS indirect. Two implants also showed angular changes but only for the PE indirect technique versus the VPS indirect technique (P < .05). CONCLUSION: The snap-on VPS indirect impression technique using a stock tray, which has the advantages of being clinically convenient and eliminating repositioning after removal of the impression, resulted in dimensional accuracy similar to that achieved with the PE direct technique. PMID- 15346749 TI - Immediate loading with fixed screw-retained provisional restorations in edentulous jaws: the pickup technique. AB - PURPOSE: This article describes (a) an immediate loading technique in the treatment of edentulous arches with screw-retained provisional restorations and (b) the effort to determine whether the described technique is compatible with the predictable achievement of osseointegration. MATERIALS AND METHODS: Eight patients with either 1 or 2 edentulous arches were treated. A diagnostic tooth arrangement was carried out for each patient and was then duplicated twice to fabricate a provisional template and a surgical guide. Six to 10 solid-screw ITI implants were placed around the dental arch to reach the first molar regions. On the same day, all patients received splinted metal-free screw-retained provisional restorations according to the pickup technique. The provisional prostheses were retrieved every 2 weeks during the healing phase. RESULTS: Seventy-eight implants were placed in 11 edentulous arches. Two implants were not immediately loaded because of inadequate primary stability. In an 8- to 20-month follow-up period (mean 14 months), two 8-mm implants were lost after 5 weeks of functional loading, resulting in an overall survival rate of 97.4%. All implants were assessed by resonance frequency analysis. After 4 months of functional loading, the mean implant stability quotient was 60+/-4.1 units (range 51 to 72 ISQ units) for maxillary implants and 65+/-6.5 units (range 47 to 74 ISQ units) for mandibular implants. DISCUSSION AND CONCLUSION: The immediate loading of implants placed in edentulous arches with screw-retained 1-piece (cross-arch) provisional restorations does not appear to jeopardize the achievement of osseointegration. Neither the metal-free design of the provisional prostheses nor the removal of the provisional prostheses during the healing phase adversely affected osseointegration. The pickup technique for immediate provisionalization represents a reproducible treatment option. PMID- 15346750 TI - Immediate provisional restoration of Osseotite implants: a clinical report of 18 month results. AB - PURPOSE: The purpose of this study was to assess the survival rates and interproximal bone levels for Osseotite implants that were restored with fixed provisional crowns without occlusion immediately after implant placement. MATERIALS AND METHODS: Ninety-three implants were placed in 38 partially edentulous patients. All implants were immediately restored with prefabricated abutments and cement-retained provisional crowns without centric or eccentric occlusal contacts. The implants were restored with definitive restorations approximately 8 to 12 weeks after implant placement. All patients included in the study were followed for at least 18 months after implant placement (average 20.3 months). RESULTS: Seventy-seven of the 93 implants satisfied the inclusion criteria. Seventy-five implants became osseointegrated. The overall survival rate was 97.4%. Radiographic bone loss 18 months after implant placement (the mean of both interproximal surfaces) was 0.76 mm. The exact binomial confidence interval was 0.32% to 9.07%. For the exact binomial test with the null hypothesis proportion = .05, P was .3334 and was not statistically significant. DISCUSSION: Immediate nonocclusal loading of single-unit dental implants differs from immediate loading of multiple, splinted implants. Unsplinted, restored implants without occlusal loading may still be subject to lateral and occlusal loads secondary to the proximate location of the food bolus. Immediate restoration of dental implants significantly reduces treatment time and may be beneficial in reducing the morbidity associated with loss of teeth, contraction of the alveolus, and loss of interdental papillae associated with the traditional method of treatment following tooth loss. CONCLUSIONS: The results of this study suggest that immediate restoration of Osseotite implants can be accomplished with results that are similar to the results obtained with the traditional 1- or 2-stage surgical, unloaded healing protocols. PMID- 15346751 TI - Assessment of surgical skills in implant dentistry. AB - PURPOSE: This study aimed to (1) compare 2 scales in the assessment of first stage implant surgery, (2) assess the interrater reliability of these scales, and (3) compare self-assessment with observer assessment. MATERIALS AND METHODS: Twenty-three patients underwent first-stage implant surgery. One assessor, an experienced dental surgeon, assisted and supervised the operator, while the second, a postgraduate trained in assessment, observed the procedure closely. The assessment scales consisted of a checklist and a global rating scale. RESULTS: A significant correlation was found between the checklist and the global rating scale scores (r = 0.47, P = .002). The British Standards Reproducibility Coefficients were 2.5 (checklist) and 7.4 (global rating scale) for interrater reproducibility and 7.0 (checklist) and 12.6 (global rating scale) for self assessment versus assessor reproducibility. Finally, analysis of the intraclass correlation coefficients between the assessors (0.74 and 0.64 for the checklist and the global rating scale, respectively) and between the surgeons' and trainers' scores (0.09 for the checklist and 0.18 for the global rating scale) showed a much weaker agreement for the latter. DISCUSSION: There was good correlation between scores using the 2 different methods of assessment. The interrater reliability was substantial for both scales. However, training of assessors to ensure higher levels of interrater reliability may be necessary. These results also demonstrated the inability of some surgeons to assess their performance accurately. CONCLUSION: Both the checklist and the global rating scales provided useful assessment data, and both were considered of value by the assessors and surgeons in providing feedback. The development of assessment and self-assessment skills in implant surgery is necessary if we are to establish a culture of commitment to lifelong learning. PMID- 15346752 TI - Soft tissue healing around implants placed immediately after tooth extraction without incision: a clinical report. AB - PURPOSE: The purposes of the present study were to evaluate implants placed immediately after tooth extraction without incision or primary flap closure and to observe the peri-implant soft tissue healing. MATERIALS AND METHODS: Fifteen patients (9 men and 6 women) aged 31 to 54 years were included in this study. Each patient had a tooth that required extraction, and each had at least 4 mm of bone beyond the root apex. Teeth with multiple roots were excluded from this study. After tooth extraction, the implants were immediately placed without incision or flap elevation. Implant sites showing bone fenestrations, bone dehiscences, or peri-implant bone defects exceeding 2 mm were excluded from this study. In these cases, a standard guided bone regeneration procedure with a surgical flap elevation was used. The second-stage surgical procedure was performed 6 months after the first procedure. The following clinical parameters were evaluated at the time of implant placement and at second-stage surgery: levels of mesial and distal papillae, width of keratinized mucosa, position of mucogingival junction relating to the surrounding tissues, and peri-implant radiolucency and marginal bone loss, which were evaluated radiographically. RESULTS: The postsurgical healing period was uneventful for all patients. Soft tissue closure over the implant sites was achieved in 1 to 3 weeks after surgery at all sites. At second-stage surgery, no peri-implant bone defects were observed or detected by probing around all the experimental implants. The soft tissue anatomy was considered clinically acceptable in all patients. DISCUSSION AND CONCLUSION: Successful osseointegration and complete bone healing were observed for all patients. The soft tissue healing and morphology were satisfactory; additional mucogingival surgery was not required before definitive prosthetic rehabilitation. PMID- 15346753 TI - Alternative bone expansion technique for immediate placement of implants in the edentulous posterior mandibular ridge: a clinical report. AB - PURPOSE: The aim of this study was to evaluate the effectiveness of a split-crest bone augmentation technique performed for immediate implant placement in thin edentulous posterior mandibular ridges. MATERIALS AND METHODS: In the present study, 125 implants were placed in posterior mandibular ridges of 30 patients. The mandibular buccal walls were split, expanded, and grafted with a combination of platelet-rich plasma and Cerasorb. The split segments were held in place by cortical bone screws. Before loading, periodontal status was checked, implant stability was evaluated with the Periotest, and panoramic radiographs and computerized tomography scans were obtained. Second-stage surgery (cover screw removal and healing abutment placement) was performed after 3 to 4 months. RESULTS: All implants osseointegrated successfully and underwent loading after 4 months. Optimal healing occurred 3 to 4 months earlier than the usual 6 to 9 months required, and no lip paresthesia was noted. DISCUSSION: Although onlay inlay grafts, sandwich osteotomies, guided bone regeneration, piezoelectricity, and alveolar distraction have been indicated for augmentation in the posterior mandibular region, each of these techniques involves risks and complications. The crest-splitting bone expansion technique enables single-stage immediate implant placement and lateral ridge augmentation in thin crests and may prevent neurosensorial deficiencies. CONCLUSIONS: The split-crest surgical technique is a valid reconstructive procedure for sharp posterior mandibular ridges. If performed using platelet-rich plasma and Cerasorb, it can shorten the osseointegration period. PMID- 15346754 TI - The effect of sinus membrane perforation and repair with Lambone on the outcome of maxillary sinus floor augmentation: a radiographic assessment. AB - PURPOSE: The present study compared the regenerative outcome of sinus graft procedures in a group of patients who underwent the repair of an intraoperatively diagnosed sinus membrane perforation to that of a group of patients without sinus membrane perforations. MATERIALS AND METHODS: A sinus floor augmentation procedure was performed in 73 sinuses in 63 patients. In 28% of these sinuses a significant (> 5 mm) membrane perforation was observed intraoperatively. In these cases, the perforation was sealed with a freeze-dried human lamellar bone sheet, and the grafting procedure was carried out as planned. The following parameters were measured on panoramic radiographs immediately postoperatively and at the 6- and 24-month follow-up examinations: (1) the distance between the occlusal edge of the implant and the preoperative sinus floor, (2) the distance between the occlusal edge of the implant and the postoperative sinus floor, and (3) the distance between the occlusal edge of the implant and the alveolar crest. RESULTS: The patients whose sinus membranes were perforated experienced no complications. No statistically significant differences were found between the 2 groups in the parameters measured. DISCUSSION: Lambone was used in all cases in the present study. In no case did the sinus augmentation procedure have to be abandoned. CONCLUSION: It can be concluded that membrane elevation must be carefully executed to avoid membrane perforation, but that if it occurs, it is still possible to continue the procedure safely after repair. PMID- 15346755 TI - The extremely resorbed mandible: a comparative prospective study of 2-year results with 3 treatment strategies. AB - PURPOSE: The aim of this prospective clinical study was to compare the clinical and radiographic results of 3 modes of implant treatment in combination with an overdenture in patients with extremely resorbed mandibles. The 3 treatment strategies used were a transmandibular implant, augmentation of the mandible with an autologous bone graft followed by placement of 4 endosseous implants, and the placement of 4 endosseous implants only. MATERIALS AND METHODS: Sixty edentulous patients met the inclusion criteria and were assigned according to a balanced allocation method to 1 of the 3 groups. Postoperative complications, implant survival, periodontal indices, change in mandibular bone height, and prosthetic complications were assessed during a 2-year evaluation period. RESULTS: During the evaluation period significantly more implants were lost in the transmandibular implant and the augmentation groups compared to the group with endosseous implants only. Except for the Bleeding Index and the Periotest values, the periodontal parameters did not differ significantly among the groups. In all 3 groups, there was no significant bone loss at most locations. Minimal prosthetic retreatment was necessary. DISCUSSION: Although implant loss is a frequently used outcome measure for success, the necessity of retreatment seems to be of more relevance for both the patient and the clinician. CONCLUSIONS: The results of this study suggested that patients with extremely resorbed mandibles can be treated with implants alone in this patient population. PMID- 15346756 TI - A comparison between cutting torque and resonance frequency in the assessment of primary stability and final torque capacity of standard and TiUnite single-tooth implants under immediate loading. AB - PURPOSE: In this study, standard Branemark System implants and Branemark TiUnite implants were evaluated regarding primary stability and placement torque. The correlation between placement torque and primary stability as well as the influence of implant design on placement torque and primary stability were examined. MATERIALS AND METHODS: Twelve patients who presented bilateral loss of either maxillary lateral incisors or premolars were treated with 24 immediately loaded dental implants. Each patient received 1 standard 3.75 x 13-mm Branemark System implant and one 3.75 x 13-mm TiUnite Mk III implant. One at a time, each implant was connected by a transducer to an Osstell machine that automatically translated a resonance frequency value for the implant into an implant stability quotient value. Osseocare equipment was used to measure the placement torque for both types of implants. RESULTS: Statistical analysis showed higher mean values for standard implants in relation to placement torque and resonance frequency values. DISCUSSION: There was no overall correlation between placement torque and resonance frequency values; this finding supported previous studies. CONCLUSION: Stability was shown to be higher for the standard implants. Mean values of torque resistance were higher for the standard implants than for the TiUnite implants. Implant design appeared to influence primary stability and placement torque. PMID- 15346757 TI - Proximal tibia bone harvest: review of technique, complications, and use in maxillofacial surgery. AB - Autologous bone grafts for alveolar ridge augmentation are the gold standard for restoring atrophic residual ridges in preprosthetic surgery. Many indications, donor sites, and techniques have been reported. The purpose of this article is to review the anatomy, surgical technique, and potential complications associated with proximal tibia bone harvest. A consecutive series of 44 patients who underwent proximal tibia bone graft harvest between 2000 and 2003 was studied by retrospective chart review. Five major and 7 minor complications were observed; overall morbidity was low. A significant amount of corticocancellous bone may be harvested from the proximal tibia with minimal morbidity. PMID- 15346758 TI - Cross-sectional study of the factors that influence radiographic magnification of implant diameter and length. AB - PURPOSE: To study the factors that influence radiographic magnification of implant diameter and length. MATERIALS AND METHODS: The dental records and panoramic radiographs of 80 patients with 210 dental implants treated with implant-supported prostheses at Bundang Jesaeng Hospital in South Korea from January 2000 through February 2003 were reviewed. The panoramic radiographs were developed under standardized conditions. The patient's gender and the anatomic locations of implants were identified from the dental records. To prevent bias, a blinded investigator measured implant diameter and length on a panoramic radiograph. To evaluate intra-examiner variability, the intraclass correlation coefficient (R(I)) was calculated. The Mann-Whitney rank-sum test and the Kruskal Wallis test were used to determine the statistical significance of the difference between actual length and radiographic length. RESULTS: The intraclass correlation coefficients (R(I)) were 0.83 for diameter and 0.87 for length. There was no statistically significant difference in length in regard to gender (P = .08). Magnification of diameter did differ on the basis of gender (P = .03; 25% magnification in radiographs of women; 20% in men). No difference in diameter was found in regard to anatomic location (P = .51), however, while evidence of difference in length in regard to anatomic location was found (P = .01). DISCUSSION: Radiographic magnification of implant dimensions in diameter and length can have different influencing factors. CONCLUSIONS: This study found that radiographic magnification of implant diameter was influenced by gender, whereas radiographic magnification of implant length was influenced by anatomic location. Each anatomic location had a different amount of radiographic magnification for implant length. PMID- 15346759 TI - Crestal bone resorption 5 years after implant loading: clinical and radiologic results with a 2-stage implant system. AB - PURPOSE: The aim of the present study was to assess crestal bone resorption 5 years after loading by conducting a clinical and radiographic evaluation of 112 Frialit-2 implants consecutively placed in 51 patients from January 1994 through June 1994. MATERIALS AND METHODS: All implants were placed in the same private practice clinic by the same surgeon. Clinical assessment included plaque score monitoring, bleeding on probing, probing depth, type of occlusion, and prosthetic adaptation. Intraoral radiographs were taken and compared using suitable software to accurately measure peri-implant bone resorption. RESULTS: The survival rate of the implants was 100%. Plaque was present on 47 (42%) implants. Bleeding on probing was detectable at 17 implants (15.5%). Probing depth was > 5 mm for 5 implants (4.5%). Crestal bone resorption was > 3 mm for 32 implants (28.6%); the average observed crestal bone resorption was 2.17+/-1.6 mm. DISCUSSION: The survival rate of the implants may be the result of the relatively short functional period as well as the strict and frequent clinical evaluations associated with oral hygiene procedures during the supportive periodontal therapy. CONCLUSION: The results suggest that with strict plaque control, and provided that the patient follows a regular program of supportive therapy, crestal bone resorption around a 2-stage implant system may be limited. PMID- 15346760 TI - Functional and esthetic rehabilitation after mandibular resection in a child using a tooth/implant-supported distraction device: a case report. AB - After resection and autogenous bone grafting in infancy and childhood, hypotrophy of the concerned jaw can often be observed. A 6-year-old male patient with osteogenic sarcoma was treated with partial resection of the mandible from the left first molar to the right first premolar. The aim was to rehabilitate the patient functionally and esthetically as he grew. At the age of 16, after the placement of 4 Branemark System implants in the consolidated autogeneous bone graft, the patient was treated with horizontal distraction osteogenesis. A new and unconventional type of individual tooth/implant-supported distraction device was used to lengthen the mandible by 16 mm. The implants placed to support the device were later used for prosthodontic rehabilitation. Progress in bone reconstruction, plastic coverage, and implant dentistry, as well as distraction osteogenesis, have enabled the compensation of functional and esthetic impairments caused by resection, especially in infants and young children. PMID- 15346761 TI - Identification of LZP gene from Mus musculus and Rattus norvegicus coding for a novel liver-specific ZP domain-containing secretory protein. AB - Zona pellucida (ZP) domain has been recognized in a number of receptor-like eukaryotic glycoproteins, which involved in many important biological processes, such as signal transduction, development, differentiation and so on. Here we report the identification of Mus musculus and Rattus norvegicus orthologues of Homo sapiens LZP gene which codes for a novel ZP domain-containing protein. Sequence analysis revealed that human, rat and mouse LZP proteins are highly conserved. Mouse LZP gene has two transcripts, 2.4 and 2.8 KB long respectively, coding for identical protein. Mouse LZP mRNA is expressed specifically in hepatocytes. Our data also showed that mouse LZP localizes mostly on nuclear envelope, and at the same time, it can be secreted into blood in a truncated form. PMID- 15346762 TI - CDNA cloning and characterization of the Ve homologue gene StVe from Solanum torvum Swartz. AB - Verticillium wilt is a disastrous disease causing significant yield losses of many crops. Isolation of verticillium wilt resistance gene is a fundamental work for controlling this disease through genetic engineering. In this report, we describe the cloning and characterization of a Ve like gene (StVe) from Solanum torvum Swartz. The nucleotide sequence of StVe is 3640 bp long with an open reading frame of 3414 bp encoding a protein precursor of 1138 aa. Sharing high homologies to tomato verticillium wilt disease resistance genes Ve1 and Ve2, the leucine rich (15.89%) protein StVe has a calculated molecular weight of 126.48kDa with an isoelectric point of 5.62. It possesses a hydrophobic N-terminal signal peptide of 20 aa and 38 predicted leucine-rich repeats containing 32 potential N glycosylation sites (28 being significant). Fifty-seven predicted phosphorylation sites (36 for S, 8 for T and 13 for Y) distribute in StVe protein. A PEST-like sequence and a mammalian endocytosis signals YCVF are found within the C-terminal region. The C terminus of StVe concludes with the residues KKF similar to the KKX motif that confers endoplasmic reticulum localization in plants as well as mammals and yeast. The sequence analysis of the StVe gene implies that the StVe is a potential verticillium wilt disease resistance gene encoding a cell surface like receptor protein. PMID- 15346763 TI - Nucleotide sequence of a portion of the camphor-degrading gene cluster from Rhodococcus sp. NCIMB 9784. AB - Rhodococcus sp. NCIMB 9784 is a camphor-degrading Gram-positive organism originally isolated from activated sewage sludge. A 5.4kbp portion of a proposed camphor degradation gene cluster from this organism was cloned and its nucleotide sequence determined. Four open reading frames (ORFs) were identified encoding proteins possibly involved in camphor metabolism; sequence alignment of the translation products suggested that the ORFs encode for a ferredoxin reductase, acyl-CoA ligase, epimerase and an acyl-CoA dehydrogenase. The last three activities are thought to be involved in the poorly understood late stage of camphor degradation. Our findings are entirely consistent with the proposed formation of a branched 9-carbon acid intermediate (3,4,4-trimethyl-5-oxo-trans-2 hexenoic acid) which has been isolated from the fermentation broth of camphor grown cells. PMID- 15346764 TI - Medicago truncatula Mt-ZFP1 encoding a root enhanced zinc finger protein is regulated by cytokinin, abscisic acid and jasmonate, but not cold. AB - A Medicago truncatula zinc finger protein cDNA (Mt-ZFP1) was isolated from a M.truncatula seedling cDNA library using RT-PCR product as a probe. The predicted amino acid sequence of Mt-ZFP1 is over 79% similar to S-SCOF-1 from soybean, a novel cold-inducible zinc finger protein involved in cold stress signal transduction mediated by abscisic acid (ABA). The secondary structure of Mt-ZFP1 protein was almost identical to that of S-SCOF-1. Mt-ZFP1 also contained a typical C2H2-type zinc finger domain and a putative nuclear located signal. RNA gel blot hybridization demonstrated that the Mt-ZFP1 gene was actively expressed in roots, with a lower abundance in leaf and stem tissues. Cold treatment did not induce the expression of Mt-ZFP1 in either leaves and stems or roots. Exogenous application of cytokinins marginally increased the accumulation of Mt-ZFP1 mRNA, while ABA and jasmonate treatments decreased the levels of Mt-ZFP1 mRNA. DNA gel blot analysis demonstrated that Mt-ZFP1 is present as a single copy gene in the M. truncatula genome. These data suggest that Mt-ZFP1 is a novel zinc finger protein with different physiological functions to that of S-SCOF-1. The similar cold-inducible factor like S-SCOF-1 might not exist in M. truncatula. PMID- 15346765 TI - Cloning and characterization of a novel avirulence gene (arp3) from Xanthomonas oryzae pv. oryzae. AB - A novel avirulence gene was cloned from Xanthomonas oryzae pv. oryzae strain PX0339, which is the standard representative of the Philippines race 9a. The full length gene spans 2118 bp and encodes a protein of 705 amino acids. BLAST search in NCBI indicated that the gene belongs to avrBs3 gene family, and designated arp3 (AvrBs3-related protein 3, arp3). The central region of the arp3 contains only 5.5 copies of 102bp repeats, the smallest copy number of repeats found in avrBs3 gene family by now. Together with the repeats is heptad repeats, resembling leucine zippers. Three functional nuclear localization signals and an acidic activation domain are also found in the C-terminal region. However, the arp3 lacks of two segments in its N-terminal region, which is unique in avrBs3 gene family. Southern blotting data showed that the arp3 is present as a single copy in genomic DNA of PX0339 and locus in plasmid clone. The arp3 could be expressed in vitro in Escherichia coli BL21 and a 128kDa fusion protein was detected by Western analysis. PMID- 15346766 TI - Cloning and overproduction of the rpoZ gene encoding an RNA polymerase omega subunit from a deep-sea piezophilic Shewanella violacea strain DSS12. AB - We have cloned the rpoZ gene, encoding RNA polymerase omega protein, by PCR approach from the deep-sea piezophilic and psychrophilic bacterium, Shewanella violacea strain DSS12. The cloned gene, 285bp in length, was found to encode a protein consisting of 94 amino acid residues with a molecular mass of 10,327 Da. Significant homology was evident comparing the RpoZ protein of S. violacea with that of Shewanella oneidensis (69% identity), Vibrio cholerae (65% identity), Escherichia coli K-12 (64% identity) and Haemophilus influenzae (61% identity). From the Northern blot analysis, S. violacea rpoZ gene was expressed constitutively under pressure conditions of 0.1, 30 and 50MPa. We constructed expression plasmid to overproduce the RpoZ protein and transformed into E. coli JM109 as a host of overproduction. Upon induction, the recombinant protein encoded by plasmid pQrpoZ was overexpressed and purified using Ni2+ affinity column. PMID- 15346767 TI - Analysis of a 108-kb region of the Saccharopolyspora spinosa genome covering the obscurin polyketide synthase locus. AB - A 108-kb genomic DNA region of Saccharopolyspora spinosa NRRL 18395, producer of the agriculturally important insecticidal antibiotics spinosyns, has been cloned, sequenced and analyzed to reveal clustered genes encoding a type I polyketide synthase (PKS) complex. The genes for the PKS are flanked by genes encoding homologs of enzymes that are involved in the urea cycle, valine, leucine and isoleucine biosynthesis and energy metabolism. While the disruption of the PKS genes by insertional inactivation was not expected to abolish the production of spinosyns, no differences were found in the antibacterial, antifungal, or insecticidal activities either of the parental and the knockout mutant strains under the growth conditions tested. Deduction of the most likely structure of the polyketide core of the cryptic metabolite, termed obscurin, from the predicted modules and domains of the PKS suggests the formation of a highly unsaturated substituted C22 carboxylic acid that might undergo further processing after its release from the PKS. PMID- 15346768 TI - Dampable waves along nucleic acid sequences mediating nucleotides' interactions. AB - This work studied the relationship of any two nucleotides in genomic sequences, coding sequences and full-length cDNAs. We made a statistical hypothesis that there exist no interactions between any two nucleotides in sequences, therefore, a hypothetical combination distribution of two nucleotides is considered and the difference between the hypothetical combination distribution and the actual distribution is used to measure the average interaction between the two nucleotides. As a result, we found that the interactions between any two nucleotides are clearly and closely related with dampable wavelike patterns along the sequences. Based on the results we daringly make some hypotheses on several biological topics. Further, studies on the wave may provide new clues for gene prediction and genome structure study. PMID- 15346769 TI - Molecular cloning and characterization of SGT1.2, a novel splice variant of Homo sapiens SGT1. AB - SGT1.2, a novel splice variant of Homo sapiens SGT1 was isolated from a human fetal brain cDNA library. This cDNA is 1404 bp and contains an open reading frame from 68 to 1165 encoding a putative protein of 365 amino acids (SGT1.2) that share 90% identities to Homo sapiens SGT1, suppressor of G2 allele of SKP1 at protein level. RPS-BLAST searching revealed that SGT1.2 have a TPR domain, a p23 domain, a SGS domain and a CS domain. According to the search of the human genome database, SGT1.2 was mapped to human chromosome 13q14.13. Reversed transcription polymerase chain reaction analysis indicated that it widely expressed in human adult tissues. PMID- 15346770 TI - Cloning and identification of a novel human gene PDLIM5, a homolog of AD associated neuronal thread protein (AD7c-NTP). AB - A novel human gene cDNA was successfully cloned from the human fetal brain cDNA library constructed by our lab, and this gene was termed PDLIM5 after acquiring the agreement of HUGO. BLASTX searching revealed that the hypothetical protein is a homolog of AD-associated neuronal thread protein (AD7c-NTP), which is over expressed in Alzheimer disease (AD) beginning early in the course of disease, and over-expression of the AD7c-NTP gene would cause neuritic sprouting and cell death. SMART analysis showed that both our predicted protein and AD7c-NTP comprise BCL domain (only contains BH1 and BH2 regions). RT-PCR experiment revealed that the expression level of PDLIM5 in brain, skeletal muscle, prostate, colon and leukocyte is obviously higher than that in other tissues. PMID- 15346771 TI - Genomic organization of mouse desmocollin genes reveals evolutionary conservation. AB - Desmosomal cadherins are a family of calcium regulated proteins involved in the formation of desmosomes, a type of cell junction important in maintaining cell adhesion and tissue stability. The desmosomal plaque consists of members of the desmosomal cadherin, plakin and armadillo family of proteins. Desmosomal cadherins are transmembrane glycoproteins that interact with desmosomal cadherins of the adjacent cells via their extracellular repeat domains and are divided in two subfamilies, the desmogleins (Dsg) and the desmocollins (Dsc). On the cytoplasmic side, the cadherins connect to the intermediate filament (IF) network indirectly by interacting with plakin and armadillo proteins. Here, we report the elucidation of the genomic structure of two mouse desmocollin genes, Dsc2 and Dsc3. Interestingly, at the genomic level, desmocollins show a higher degree of similarity to the classical cadherins, such as E-cadherin, than to the desmogleins. PMID- 15346772 TI - cDNA cloning of translationally controlled tumor protein/histamine releasing factor (TCTP/HRF) from the intertidal harpacticoid copepod Tigriopus japonicus. AB - We synthesized a cDNA library from the intertidal copepod Tigriopus japonicus, converted it to phagemids and sequenced expressed sequence tags (ESTs). Of these, Tigriopus translationally controlled tumor protein/histamine releasing factor (TCTP/HRF) was further characterized. The Tigriopus TCTP/HRF gene encoded 172 amino acid residues and showed high similarity to Drosophila but moderate similarity to other annelids (e.g. Brugia, Wuchereria and C. elegans). The Tigriopus TCTP/HRF gene appeared in the same clade as the annelids. Here, we describe the analysis of the Tigriopus TCTP/HRF gene. PMID- 15346773 TI - Arsenic sorption by carbonate-rich aquifer material, a control on arsenic mobility at Zimapan, Mexico. AB - Arsenic retention by carbonate-rich aquifer material at Zimapan, Mexico, was studied to gain insight into the processes controlling arsenic mobility in natural systems. Batch experiments showed that retention of soluble As (V) on carbonate-rich aquifer material was 35.3-90% in the pH range of 7-9 found in Zimapan natural water. Chemical and mineralogical compositions, point of zero charge (PZC), and experimental arsenic retention at various pH of three composite samples of limestone (M1, M2, M3) showed that sorption (adsorption and coprecipitation) may be one of the main processes controlling arsenic mobility in the Zimapan As-polluted aquifer. The PZC values approximately corresponded to the PZC reported for the main minerals present in each sample: hydrous ferric oxides (M1), calcite (M2), and the range from hydrous ferric oxides and calcite (M3). The chemical and mineralogical compositions of each sample explain the obtained PZC values. Experimental and modeled arsenic retention at various pH values on sample M1 corresponded to reported arsenic adsorption onto hydrous ferric oxides. Coprecipitacion of complex Ca arsenates or arsenic adsorption onto calcite or clay minerals could be the main processes of arsenic retention on samples M2 and M3. Groundwater flow through the granular, carbonate-rich, shallow aquifer may decrease the water As content as a result of these interactions. A remediation method based on the promotion of polluted water flow into the shallow aquifer could be developed from these results. PMID- 15346774 TI - Assessment of the pollution status of alluvial plains: a case study for the dredged sediment-derived soils along the Leie River. AB - The concept of an integrated multifunctional river management is gaining importance. For major rivers, restoring the contact between rivers and alluvial plains is an important goal, as riparian areas have a specific role for several riverine processes. However, former and current human activities are an obstruction or a limitation for river restoration. We studied the influence of former dredging activities along the river Leie on the alluvial plain quality. A soil survey and an archive query for reconstructing the history of dredging operations were conducted simultaneously. The geographical impact expressed as topographical changes and covering of the original soil profile and related processes and biota was large. The pollution status of dredged sediment-derived soils was found to be far from negligible: concentrations of Cd, Cr, and Zn were, in 10% of the cases, higher than 20, 480, and 2800 mg kg(-1) DM, respectively. Both agriculture and nature rehabilitation on dredged sediment-derived soils can only be accepted after profound risk assessment, and management should focus on ecological risk reduction. Results indicate the importance of soil quality assessment in alluvial plains for an integrated river management, rather than a priori assuming pristine soil conditions. The collected "off-line" sediment data can be used as a reconstruction of past sediment pollution, especially when long term sediment monitoring programs are not available. PMID- 15346775 TI - Assessment of heavy metal contamination in roadside surface soil and vegetation from the West Bank. AB - Concentrations of heavy metals (Pb, Cd, Cu, Zn, Fe, Mn, Ni, and Cr) were investigated in roadside surface soil and the common perennial herb inula (Inula viscosa L., Compositae). Samples were collected at different distances (0-200 m) perpendicular to a main road that connects two main cities in the West Bank. Average concentrations of metals in soil samples were: Pb, 87.4; Cd, 0.27; Cu, 60.4; Zn, 82.2; Fe, 15,700; Mn, 224; Ni, 18.9; and Cr, 42.4 microg x g(-1). In plant leaves, concentrations were: Pb, 7.25; Cd, 0.10; Cu, 10.6; Zn, 47.6; Fe, 730; Mn, 140; Ni, 4.87; and Cr, 7.03 microg x g(-1). Roadside contamination was obvious by the significant negative correlations between concentrations of metals in soil and plant samples and distance from road edge. Only cadmium concentrations in soil and plant samples were not associated with roadside pollution. Roadside contamination in plants and soil did not extend much beyond a 20 m distance from road. I. viscosa reflected roadside contamination better than soil and their metal concentrations showed much less fluctuations than those in soil samples. Washing plant leaves decreased Pb and Fe concentrations significantly, indicating a significant aerial deposition of both. I. viscosa can be considered as a good biomonitor for roadside metal pollution. PMID- 15346776 TI - Effects of pig slurry on the sorption of sulfonamide antibiotics in soil. AB - Sorption of p-aminobenzoic acid (pABA) and five sulfonamide antibiotics to loess Chernozem topsoil amended with varied additions of pig slurry was investigated in batch trials. In unfertilized soil, partition coefficients (Kd) of sulfonamides ranged from 0.3 to 2.0. Strong sorption nonlinearity (1/n = 0.5 to 0.8) was best fitted by the Freundlich isotherm (R2 = 0.7 to 1.0) and was indicative for specific sorption mechanisms. Adsorption to pig slurry was much stronger, and nondesorbable portions were increased compared with soil. However, in a mixture of soil and slurry (50:1 w/w), sorption of the antibiotics was significantly decreased at a lower concentration range of pABA and the sulfonamides. This was attributed to competitive adsorption of dissolved organic matter (DOM) constituents from manure. An increase in pig slurry amendment resulted in increased total organic matter, DOM concentration, and ionic strength, but pH decreased. As a result, the nonadsorbed portions of pABA, sulfanilamide, and sulfadiazine (logD(ow) < -0.4) ranged from 47% to 82% of the applied concentration in the differently manured substrates. Dissolved fractions of the antibiotics reached a maximum at a soil-slurry ratio of 9:1 and decreased with further addition of manure. This decrease was related to the formation of less effective DOM associates in solution. The adsorbed and desorbed portions of the less-polar substances--sulfadimidine, sulfadimethoxine, and sulfapyridine (logD(ow) > 0.1)--remained nearly constant in the presence of increased manure input. The pH changes caused by manure amendment strongly affected ionisation status of the latter compounds, thus resulting in increased adsorption, which compensated the mobilizing effect of DOM. It is suggested that the effect of manure be considered in test methods to determine the soil retention of pharmaceutical substances. PMID- 15346777 TI - Development of a sublethal test to determine the effects of copper and lead on scleractinian coral larvae. AB - A new sublethal toxicity test was developed in this study to measure the effect of copper and lead on the motility of coral larvae. Larval motility was significantly affected by copper and lead doses immediately after dosing. The copper EC50 values for motility of Goniastrea aspera brain coral larvae (12 h, 21 microg/L; 24 h, 16 microg/L; 48 h, 22 microg/L) were much lower than the copper LC50 values for G. aspera larval survival (6 h, 260 microg/L, and 24 h, 121 microg/L, for 5-day-old larvae and 6 h, 248 microg/L, and 24 h, 136.64 microg/L, for 6-day-old larvae) during the early part of the experiments. However, at later times, the LC50 values (48 h, 40 microg/L, for 5-day-old larvae and 48- h, 87 microg/L, for 6-day-old larvae) were similar to the EC50 values for larval motility. The lead 72-h EC50 value for G. aspera larval motility (2900 microg/L) was much lower than the lead 72-h LC50 value for larval survival (9890 microg/L). The results show that larval motility can be a useful parameter to measure in order to determine the sublethal effects of trace metals on coral larvae. PMID- 15346778 TI - Response of zooplankton and phytoplankton communities to creosote-impregnated Douglas fir pilings in freshwater microcosms. AB - Creosote has been used extensively as an industrial wood preservative for the protection of marine pilings, railway ties, and utility poles and is a common source of polycyclic aromatic hydrocarbons (PAHs) into aquatic environments. At present, there is little information by which to judge the potential for creosote leached from impregnated pilings to cause toxicity to biota in aquatic environments. The objective of the current study was to assess the effects of creosote on zooplankton and phytoplankton populations in freshwater microcosms in relation to changes in the concentration and composition of PAHs leached from creosote-impregnated Douglas fir pilings during an 83-day exposure period. The study consisted of single microcosms containing one half, one, two, three, four, and six treated pilings. Two microcosms that received untreated pilings were used as controls. The total surface area of pilings in each microcosm was normalized by adding the appropriate number of untreated pilings. Samples were collected periodically between -14 and 83 days pre- and postexposure to determine aqueous concentrations of 15 priority PAHs and to assess the response of zooplankton and phytoplankton communities. Plankton community response to creosote was analyzed using principle responses curves. Peak aqueous concentrations of sigmaPAH occurred at day 7, ranging from 7.3 to 97.3 microg/L. Zooplankton abundance decreased in all microcosms after introduction of the impregnated pilings, with the magnitude of response varying as a function of aqueous creosote concentration. Using inverse regression, a no-observed-effect concentration for the zooplankton community of 11.1 microg/L was estimated. In contrast, algal abundance and diversity increased in all treatments between 7 and 21 days and attained levels up to twice that in control microcosms. This trend most likely reflected decreased grazing pressure because of the decrease in zooplankton populations, but it may also have reflected growth stimulation resulting from exposure to various constituents within the creosote mixture. Our results indicate that creosote leached from impregnated pilings deployed under typical conditions (e.g., wharves) may cause transient toxicity to benthic or limnetic communities shortly after deployment, but this likely poses few long-term risks to aquatic freshwater plankton communities. PMID- 15346779 TI - Assessing the risks to zooplankton grazers of continuous versus pulsed cypermethrin exposures from marine cage aquaculture. AB - Survival responses of adult stages of the calanoid copepod A. tonsa during and after pulse exposures to cypermethrin were used to evaluate the environmental risks of cypermethrin treatments in the marine environment. To control sea lice, whole cages of salmon are exposed to cypermethrin over 1 h, after which the effluent is released into the environment and subsequently dissipates. Here we simulate this short presence of the chemical in the water column. The present study revealed that delayed toxicity in A. tonsa adults, here expressed as decreased survival, occurred following short term pulse exposures to cypermethrin. Persistent, sensitive responses to cypermethrin during and after pulse exposures of 24 h were similar to those reported in conventional acute tests, suggesting that damage caused by accumulated levels of cypermethrin during the first 24 h of exposure could explain long-term toxicity responses in A. tonsa. Delayed toxicity following 1 h pulse exposures to cypermethrin impaired male survival and therefore changed copepod sex ratios. The proposed delayed toxicity tests were considered sensitive and suggest potentially detrimental effects of cypermethrin on copepod populations within the vicinity of the cages. By monitoring toxic responses following pulsed exposures, delayed toxicity tests are especially suitable to assess environmental risks of poorly water-soluble substances that dissipate rapidly within the water column. PMID- 15346780 TI - Determination of laboratory and field elimination rates of polychlorinated biphenyls (Pcbs) in the freshwater mussel, Elliptio complanata. AB - Chemical elimination rate constants (k2) were determined for 41 polychlorinated biphenyl (PCB) congeners in dosed freshwater mussels, Elliptio complanta, following a 150-day laboratory depuration period. Congener-specific elimination rates were inversely dependent on the n-octanol/water partition coefficient (K(ow)) of the chemical according to the regression equation: logk2 = (-0.59 +/- 0.05) logK(ow) + (2.05 +/- 0.28) (R2 = 0.80, p < 0.001). PCB elimination rate constants in E. complanta were lower than reported for zebra mussels and green lipped mussels but similar in magnitude and K(ow) dependence to data reported for American oysters. In order to validate the laboratory-derived PCB elimination rate constants, mussels dosed with [13C]PCB153 were allowed to depurate at one of the biomonitoring stations utilized in the Detroit River Biomonitoring Program and sampled at the same time intervals as laboratory animals. The field elimination rate constant for [13C]PCB153 was significantly greater than, but within a factor of 3 of, the laboratory elimination rate constant determined for unlabeled PCB153. This similarity in estimates of k2 for labeled and unlabeled PCB153 indicates that there is relatively little error introduced by using laboratory PCB elimination rate constants to estimate PCB153 toxicokinetics in mussels deployed at this field station. Elimination rate constants determined for PCBs in this species were lower and exhibited a stronger K(ow) dependence than elimination rate constants reported for selected PAHs. This suggests that E. complanata may possess some capability for PAH biotransformation. PMID- 15346781 TI - Polychlorinated naphthalenes and polychlorinated biphenyls in benthic organisms of a Great Lakes food chain. AB - Invasion of zebra mussels, Dreissena polymorpha, and round gobies, Neogobius melanostomus, into the Great Lakes has altered the food web structure and thereby the pathways of toxic contaminants such as polychlorinated biphenyls (PCBs) and polychlorinated naphthalenes (PCNs). In this study, concentrations of PCNs and PCBs were measured in organisms of a Great Lakes benthic food chain encompassing zebra mussels. PCNs were found in all of the benthic organisms, including phytoplankton, algae, amphipods, zebra mussels, round goby, and smallmouth bass, Micropterus dolomieui. Concentrations of PCNs were greater in samples collected from the Raisin River than in samples from the St. Clair River. Biomagnification factors (BMF) for tetra- through octa-CN congeners in going from algae to zebra mussels from the St. Clair River ranged from 3 to 10. No major biomagnification of PCNs was found in round gobies, when concentrations were related to those in their prey species, zebra mussels. The biomagnification potential of PCNs appears to be similar to that of PCBs in the benthic food chain investigated in this study, despite the fact that PCNs may be metabolized by organisms higher in the food chain. Among several congeners, the BMFs of PCN congeners 35, 42, 43/45, 52/60, 58, and 66/67 were highest in round gobies. PCNs accounted for 1-22% of the total TEQs (toxic equivalents) of PCBs and PCNs in benthic organisms analyzed in this study. PCB congener 126 was the major contributor to TEQs, accounting for 72-99% of the PCB-TEQs in the food chain organisms analyzed. PMID- 15346782 TI - Temporal trends of organochlorine pesticides in prawn (Macrobrachium nipponense) from Lake Kasumigaura, Japan, during 1978-2000. AB - Temporal trends of organochlorine pesticides such as beta-hexachlorocyclohexane (beta-HCH), hexachlorobenzene (HCB), and 1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (p,p'-DDE), were determined in freshwater prawn (Macrobrachium nipponense) collected from Lake Kasumigaura in 1978 to 2000. The organochlorine pesticides elucidated a 72.4% decrease in beta-HCH, from 42 (1978) to 11.6 ng/g fat (2000); an 89.6% decrease in HCB, from 1.64 (1978) to 0.17 ng/g fat (2000); and an 87.3% decrease in p,p'-DDE, from 97.5 (1978) to 12.4 ng/g fat (1992). After organochlorine pesticide prohibition in 1971, the declining trend of beta HCH, HCB, and p,p'-DDE concentrations persisting in the prawns could be approximated by exponential equations. PMID- 15346783 TI - Effects of chlorinated solvents on four species of North American amphibians. AB - Tetrachloroethylene (PCE), a dry cleaning and degreasing solvent, can enter groundwater through accidental leaks or spills, and concentrations as high as 75 mg/L have been reported in Canadian aquifers. Amphibians in wetlands receiving contaminated groundwater may be exposed to PCE and its degradation products, but little information is available on the impacts of these compounds on indigenous amphibian species. Acute (96-h static renewal) exposures to PCE and its major degradation products, trichloroethylene (TCE) and cisand trans-dichloroethylene, were conducted on embryos of four North American amphibian species: wood frogs (Rana sylvatica), green frogs (R. clamitans), American toads (Bufo americanus), and spotted salamanders (Ambystoma maculatum). Subsequently, chronic exposures to PCE and TCE were conducted with the larvae of American toads. Both PCE and TCE were teratogenic to amphibian embryos; median effective concentrations (EC50s) for developmental deformities produced by PCE and TCE exposure for wood frogs and green frogs were 12 and 40 mg/L, respectively. Embryonic survivorship, however, was not compromised at these concentrations. American toads were less sensitive; the EC50 for developmental abnormalities was not attained at the highest test concentrations, 45 and 85 mg/L PCE and TCE, respectively. These results are pertinent in assessing the impact of groundwater pollution on an aquifer-fed wetland. PMID- 15346784 TI - Copper/metal ratios in the gills of rainbow trout (Oncorhynchus mykiss) provide evidence of copper exposure under conditions of mixed-metal exposure. AB - Previous work has suggested that the ratio of copper residues to zinc in the gills of rainbow trout may indicate short-term exposure to increased levels of waterborne copper. However, the effect of exposure to a combination of increased copper and zinc concentrates in the water column was unknown. We exposed rainbow trout to 8 +/- 2 microg L(-1), 40 +/- 2 microg L(-1) and 90 +/- 9 microg L(-1) of waterborne copper and 21 +/- 3 microg L(-1), 129 +/- 40 microg L(-1), and 202 +/- 40 microg L(-1) of waterborne zinc in a 2-factor experiments and gill copper and zinc residues were examined. Other gill parameters analyzed included the concentrations of calcium, magnesium, sodium, and potassium, liver copper and zinc concentrations and plasma copper, calcium, sodium, and potassium are also reported here. Copper residues in the gill filaments were significantly higher in the highest level of copper exposure (high Cu, 4.06 microg g(-1); low Cu 2.41 microg g(-1); 0 Cu 2.01 microg g(-1); P = 0.001), whereas no differences were seen in zinc concentrations at any treatment level. Gill sodium and plasma calcium concentrations were also decreased at the highest waterborne copper concentrations. Although copper-zinc ratios in the gills were significantly different between the highest and lowest copper treatments (P = 0.002, F = 6.59), copper-sodium and copper-magnesium ratios were more sensitive to waterborne copper exposure (P = 0.001, F = 17.91 and P = 0.002, F = 15.45, respectively). These copper-metal ratios may be better indicators of copper loading in the water column. PMID- 15346785 TI - Trace metals in tissues of resident and migratory birds from a lagoon associated with an agricultural drainage basin (SE Gulf of California). AB - With the aim of knowing Cd, Cu, Fe, Mn, Pb, and Zn concentrations in selected tissues of birds from two places at Altata-Ensenada del Pabellon lagoon in the SE Gulf of California, 39 specimens of 14 species were analyzed. Migratory birds in this study showed the highest values of Cd, Cu, and Pb in liver; Fe and Mn, in viscera; and Zn, in feathers. Concerning the resident avifauna, the highest levels of Cd, Cu, and Fe were detected in the liver, Mn in viscera; and Pb and Zn, in feathers. Regarding Cu and Fe, higher concentrations were detected in migratory avifauna, while Mn and Zn were more accumulated in resident waterfowl. In the case of Cd and Pb, both elements showed a tendency to be more accumulated in resident seabirds. Statistical comparisons showed that in carnivorous and omnivorous birds the resident component was the group with higher levels in more comparisons where mainly Cu and Fe were involved. In the rest of the metals the differences in the concentrations in the distinct tissues of migratory versus resident species were not clearly evident in one group in particular. In a few cases, carnivorous birds had higher levels of Cd than herbivorous and omnivorous birds; for the rest of the metals there was not a clear trend of metal accumulation. PMID- 15346786 TI - Contaminant exposure and reproductive success of ospreys (Pandion haliaetus) nesting in Chesapeake Bay regions of concern. AB - The Chesapeake Bay osprey population has more than doubled in size since restrictions were placed on the production and use of DDT and other toxic organochlorine contaminants in the 1970s. Ospreys are now nesting in the most highly polluted portions of the Bay. In 2000 and 2001, contaminant exposure and reproduction were monitored in ospreys nesting in regions of concern, including Baltimore Harbor and the Patapsco River, the Anacostia and middle Potomac rivers, and the Elizabeth River, and a presumed reference site consisting of the South, West, and Rhode rivers. A "sample egg" from each study nest was collected for contaminant analysis, and the fate of eggs remaining in each nest (n = 14 16/site) was monitored at 7- to 10-day intervals from egg incubation through fledging of young. Ospreys fledged young in regions of concern (observed success: 0.88-1.53 fledglings/active nest), although productivity was marginal for sustaining local populations in Baltimore Harbor and the Patapsco River and in the Anacostia and middle Potomac rivers. Concentrations of p,p'-DDE and many other organochlorine pesticides or metabolites, total PCBs, some arylhydrocarbon receptor-active PCB congeners and polybrominated diphenyl ether congeners, and perfluorooctanesulfonate were often greater in sample eggs from regions of concern compared to the reference site. Nonetheless, logistic regression analyses did not provide evidence linking marginal productivity to p,p'-DDE, total PCBs, or arylhydrocarbon receptor-active PCB congener exposure in regions of concern. In view of the moderate concentrations of total PCBs in eggs from the reference site, concerns related to new and emerging toxicants, and the absence of ecotoxicological data for terrestrial vertebrates in many Bay tributaries, a more thorough spatial evaluation of contaminant exposure in ospreys throughout the Chesapeake may be warranted. PMID- 15346787 TI - [The karyotype of Omisus caledonicus (Edwards) (Diptera, Chironomidae)]. AB - Karyotype of Omisus caledonicus with 2n = 14 is described, all chromosomes are acrocentric, heterochromatin centromere is vacuolized. The following sequence discovered: A1, A2, B1, B2, C1, C2, D1, D2, E1, F1, G1 and G2. Simple inversions are found in chromosomes A-C and G, a complex reconstruction with 3 break points are observed in chromosome D. All reconstructions are large (from 66 to 99% of bands). According to phylogenetic reconstructions for morphological characters (Saether, 1979) and data on karyotypes it may be considered that O. caledonicus is one of much more primitive species in the tribe Chironomini. PMID- 15346788 TI - [Different sensitivity of inbred mice to hepatocarcinogen ortho-aminoazotoluene may be due to differences in the negative control mechanisms of hepatocyte proliferation]. AB - A most convenient model to study mechanisms of live organism response to chemical carcinogens is tumor induction in murine liver by aminoazodyes, in particular by ortho-aminoazotoluene (OAT). We studied both early and late stages of hepatocarcinogenesis on several lines of inbred mice differing in sensibility to OAT. By means of autoradiography, we examined proliferative activity of hepatocytes obtained from the liver of sensitive (A/He, DD, SWR) and resistant to OAT AKR, CC57Br, BALB/c lines of mice, which were injected carcinogen. The level of p53, p21Cip1, bax, mdm2, cyclin G, gadd45 genes expression in the liver of mice of different lines given OAT injection was studied by multiplex PCR method. Carcinogen caused a decrease of hepatocyte proliferative activity induced by partial hypatectomy (PHE), and an increase in p53, p21Cip, bax, mdm2, and cyclin G genes within mice of A/He, DD and SWR lines. Cell fusion experiments on hepatocytes obtained from regenerating murine liver sensitive to A/He line carcinogen and given long-time OAT administrations with resting and proliferating fibroblasts of NIH 3T3 mice revealed no obvious suppression of DNA synthesis in heterokaryons. Unlike, in fusion experiments on serum-stimulated fibroblasts with hepatocytes obtained from the liver of BALB/c line mice also given OAT suppression of DNA synthesis in stimulated fibroblasts in heterokaryons was observed 15 days following PHE. These results enable us to conclude that OAT administrations break negative endogenous mechanisms of hepatocyte proliferation control in the liver of mice sensitive to carcinogenes. PMID- 15346789 TI - [Mechanisms of protective action of wheat germ agglutinin on cell growth in wheat seedling roots under salinity]. AB - Effects of 20 nM wheat germ agglutinin (WGA) on relative growth rate, mitotic index (MI) and the cell area in the root extension zone were investigated in seedling of Triticum aestivum L. under the influence of 2% NaCl. It was elucidated that pretreatment of wheat seedling with WGA prevented a salinity induced inhibition of root cell growth, and accelerated the restoration of cell growth after stress removal. The protective WGA effect on root cell growth may be due, presumably, to reorganization of phytohormone balance caused by WGA treatment, which could lead to accumulation of LAA and decrease in the ABA level. PMID- 15346790 TI - [Molecular mechanisms of regulatory action of adrenergic receptor agonists on functional activity of adenylyl cyclase signaling system of the ciliates Dileptus anser and Tetrahymena pyriformis]. AB - Adenylyl cyclase signaling system (ACS) of the higher eukaryotes involves the following main components: receptor, heterotrimeric G protein, adenylyl cyclase (AC), and protein kinase A. At present, these components have been found in cells of different species of the lower eukaryotes. Hence, the signal transduction through ACS of unicellular eukaryotes may have some features in common with those of the higher eukaryotes. We showed earlier that agonists of adrenergic receptors (ARs) regulate AC activity of ciliates Dileptus anser and Tetrahymena pyriformis. The aim of this work was to study molecular mechanisms of AR ligand action on the functional activity of different components of ACS of the ciliates. It has been shown that beta-AR antagonist [3H]-dihydroalprenolol binds membranes of the ciliates with a comparatively lower affinity than those of the higher eukaryotes (Kd for D. anser was 13.4 nM, for T. pyriformis--27 nM). Beta-AR ligands--agonist (-)-isoproterenol and antagonists propranolol and atenolol in competition manner displace [3H]-dihydroalprenolol with IC50 that are 10-100 times higher than corresponding IC50 of beta-AR of the higher eukaryotes. In the presence of GTP, the right shift of competition curves of [3H]-dihydroalprenolol displacement by isoproterenol was obtained, being most considerable in the case of D. anser. Adrenaline and isoproterenol in a dose-dependent manner stimulated GTP-binding in cell cultures of D. anser and T. pyriformis. Suramin (10(-5) M), the inhibitor of heterotrimeric G proteins, completely blocked effects of these hormones. In D. anser culture, adrenaline and isoproterenol in a dose-dependent manner, stimulated AC activity, and its stimulating effects in the presence of beta-AR blockers vanished (propranolol) or decreased to a great extent (atenolol). At the same time the effects were unchanged in the presence of alpha2-AR antagonists yohimbine and idazoxan. These data show the involvement of G protein-coupled beta AR in signal transduction induced by AR agonists in D. anser cells. In cell culture of T. pyriformis isoproterenol weakly stimulated AC activity, and its effect was completely blocked by beta-AR blockers. Adrenaline in T. pyriformis cells in a dose-dependent manner inhibited AC activity. Inhibiting effect of hormone was decreased in the presence of alpha2-AR blockers. On the basis of the obtained data we concluded that adrenaline in T. pyriformis cells inhibited AC activity through G protein-coupled receptor, being close to alpha2-AR of vertebrate animals. PMID- 15346791 TI - [Modulation of miniature inhibitory potentials in motoneurons of the turtle spinal cord by group II metabotropic glutamate receptors]. AB - The role of group II metabotropic glutamate receptors (mGluRs) in modulation of inhibitory synaptic activity was studied by intracellular recording of motoneuron miniature inhibitory spontaneous postsynaptic potentials (mIPSPs) in isolated lumbar segments of the turtle spinal cord in the medium containing TTX, CNQX, AP 5. The ratio of mIPSPs with fast and slow kinetics (83% vs 17%) is in accordance with the ratio shown for glycine- and GABA-mediated IPSP or IPSCs (Jones et al., 1988; Gao et al., 2001). In the majority of investigated motoneurons, the selective group II mGluRs antagonist EGLU (100-250 microM) increased the frequency of mIPSPs by 106.6 +/- 74.4% (n = 9) without affecting average amplitude, suggesting a presynaptic site of mGluRs action providing for the transmitter release reduction. The analysis of EGLU action on mIPSPs with different time courses (selection by half-width) showed that the frequency of inhancement of miniature inhibitory activity is caused by predominantly short duration mIPSPs (ba 84.0 +/- 18.2%; n = 9), which are probably glycineergic. However, EGLU did not influence the mIPSPs frequency under condition of GABA receptor blockade by bicuculline (20 microM). This fact suggest that group II mGluRs could modulate glycinergic transmission to the turtle spinal motoneurons on the necessary condition that GABergic system is active. PMID- 15346792 TI - [In vitro cell adhesion and integrin expression by calcium ionophore-treated mononuclear cells from patients with acute myeloid leukemia]. AB - As shown elsewhere, cultured acute myeloid leukaemia blasts acquire certain characteristics of dendritic cells upon stimulation with cytokines and calcium ionophore. The ability of leukaemia-derived dendritic-like cells to express immune costimulatory molecules and dendritic cell marker CD83 has been extensively investigated. Although migratory capacity is a major attribute of dendritic cells, the ability of in vitro modified blasts for adhesion, chemotaxis and homing remain elusive. In the present paper, we show that after stimulation with calcium ionophore acute myeloid leukaemia blasts as well as normal dendritic cell precursors demonstrate increased capacity of binding fibronectin and denatured collagen. The expression pattern of integrins on dendritic-like leukaemic cells in general closely resembles that of monocyte-derived dendritic cells, however, variation in cell properties isolated from blood of individual patients are observed. PMID- 15346793 TI - [NADPH-diaphorase localization in the radial nerve cords of the starfish Patiria pectinifera]. AB - The presence and localization of NADPH-diaphorase (NADPH-d) in the radial nerve cords of Patiria pectonifera was shown by electron histochemistry. NADPH-d positive structures were found in ectoneural and hyponeural regions of the radial nerve cord. Ultrastructural localization of NADPH-d was detected in neurons, sensory cells, supporting cells, and in the nerve plexus. The highest enzymatic activity in ectoneural region of the radial nerve cord is due, presumably, to the involvement of NADPH in sensory signal processing. PMID- 15346794 TI - [Expression of urokinase plasminogen activator, its receptor and plasminogen activator inhibitor type 1 in different types of atherosclerotic lesion in human aorta]. AB - The role of plasminogen activators in the regulation of key processes of atherosclerosis progression stays unclear. The aim of this study was to evaluate the expression of urokinase plasminogen activator (uPA), its receptor (uPAR) and the plasminogen activator inhibitor type 1 (PAI-1) in human aorta, and to balance them with the stage of atherosclerotic lesion. We have shown that uPA and uPAR in normal aorta are mostly expressed by intimal smooth muscle cells. The expression of these proteins was up-regulated in diseased aorta compared to normal artery. The most part of cells in both fatty streak and fibro-fatty lesion were monocytes/macrophages, and about 60% of these cells expressed uPA and its receptor. PAI-1 was mostly localized on the lumonal part of the aorta and in the extracellular matrix of the intima. We observed a moderate increase of PAI-1 expression in atherosclerotic lesion. Thus, our data indicate participation of plasminogen system in atherogenesis. PMID- 15346795 TI - [Changes in human burn fluid biological activity during normal burn healing]. AB - The main goal of this work was monitoring the changes occurring in human burn fluid biological activity during normal burn healing. The fluid available in the burn until healing makes a good material for controlling biochemical microenvironment of burn cells. This environment involves factors, such as extracellular matrix proteins and matrix metalloproteinases. In this work our previous studies of the influence of wound and burn fluids on the functional activity of cells were extended to include the effect of burn fluid on fibroblasts and keratinocytes, i. e. human skin cells present in the wound and involved in wound healing. It was shown that human burn fluid biological activity depends on the time that passed after burning, and on the correctness of healing. Migration of human fibroblasts becomes more intensive under the influence of such a fluid independently on the time of fluid sampling. Unlike, keratinocyte migration was inhibited by burn fluid sampled 1-3 days after burning but was enhanced by fluids sampled 6 days following burning. The obtained data are to be necessarily taken into consideration at burn treatment and also at transplantation of cells for healing of wounds of different nature. PMID- 15346796 TI - [Nuclei and nucleolar organizing regions in chromosomes of lymphocytes on different stages of periodic disease]. AB - By scanning cytomorphometry a cytological study was first performed on the behavior of nuclei and nucleolar organizing regions (NOR) in chromosomes of peripheral blood lymphocytes of healthy men and of patients with periodic disease (familial Mediterranean fever, FMF) on different stages of development, including its complication with amyloidosis. The volume and total surface of nuclei, the sum total volume and sum total surface of NOR, the mean number of NOR for one nucleus and distribution of nuclei according to NOR number were measured. It is shown that the parameters of nuclei and NOR for patients with FMF on all stages clearly and trustworthy differ from those for healthy men. They are sufficiently informative, can be successfully used in clinical practice and even serve as an early diagnostic test for amyloidosis complication. PMID- 15346797 TI - [DNA quantification in nuclei of cultivated mushroom with DAPI staining]. AB - Agaricus bisporus (Lange) Imbach is actively cultivated amphithallic basidiomycete, in which various strains are primary homothallic, heterothallic or secondary homothallic. Countings of relative nuclear DNA content by means of DAPI stain and its comparison in different strains can help to understand the mushroom's life cycle features. The authors for the first time observed change of nuclear phases in basidia of A. bisporus strains with different types of life cycle and revealed that DNA content in diploid nuclei is about 1.3 times higher than in haploid ones. The method is highly sensitive and can be used for quantitative measurings of nuclear DNA even in objects with nuclei of about 1 mkm in diameter. PMID- 15346798 TI - Healing or curing. PMID- 15346799 TI - Management of acute exacerbations of chronic obstructive pulmonary disease in the elderly: physician practices in the community hospital setting. AB - Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death and sixth most common reason that Medicare patients are hospitalized. We performed retrospective chart review on a statewide random sample of 409 Medicare patients discharged from October 1, 2000, through January 31, 2001, with a diagnosis of COPD. The most commonly performed diagnostic tests were chest radiography (96.8%) and oxygenation assessment (94.9%). The most common treatments provided were inhaled short-acting beta-agonist bronchodilators (98.5%) and oxygen (94.4%). Antibiotics (89.0%) and systemic corticosteroids (85.1%) were prescribed less frequently. The median length of stay was 5 days. The readmission rate was 27.4% (CI, 23.1-32.0) at 30 days and 43.0% (CI, 38.2 47.9) at 180 days. The in-hospital mortality rate was 1.7% (CI, 0.7-3.5) and the 180-day mortality rate was 23.7% (CI, 19.7-28.1). The morbidity and mortality associated with acute exacerbations of COPD remain high. There are opportunities to improve quality of care for this condition. PMID- 15346800 TI - Spontaneous rupture of liver in a non-pregnant patient: case report. AB - Spontaneous rupture of the liver during pregnancy associated with preeclampsia is an uncommon and frequently fatal complication. A case of a 61-year-old non pregnant female is described here who took estrogen replacement for 16 years and presented with spontaneous rupture of the right lobe of the liver with hemoperitoneum. The underlying cause of the rupture was not clear. PMID- 15346801 TI - Radio frequency fistulotomy--a novel modification of the conventional technique in low anal fistula. PMID- 15346802 TI - Oklahoma and the issue of tobacco. AB - Tobacco use, particularly cigarette smoking, continues to be the leading cause of preventable morbidity and mortality in the United States. An important public health issue, this has been the focus of much study. Several organizations have issued statements and recommendations for handling this issue. Reducing tobacco use is a key component of Healthy People 2010, the national action plan for improving the health of all Americans. The Surgeon General, Agency for Healthcare Research and Quality (AHRQ) and the U.S. Preventive Services Task Force (USPSTF) have put forth valuable guidelines concerning preventive services in the primary care setting, and programs such as Putting Prevention into Practice (PPIP) and state based initiatives have aimed to implement them. States that have taken these messages to heart and acted upon them to fully implement these preventive measures and interventions have had significant success, most notably Oregon, California and Massachusetts. In less than a decade, the Massachusetts Tobacco Control Program has evolved into one of the leading public health initiatives of our time. Perhaps by learning from their successes we can bring about a similar change in the state of Oklahoma. PMID- 15346803 TI - Ben H. Nicholson, MD: a remarkable physician. PMID- 15346804 TI - Doctors' new tool to fight lawsuits: saying 'I'm sorry.' Malpractice insurers find owning up to errors soothes patient anger. 'The risks are extraordinary'. AB - A lot of attention and energy has been spent over the past several years on reducing the amount of settlements and awards in malpractice cases. Of course these are important issues, but the best situation for physicians is not to be sued at all. Therefore, the medical community needs to start focusing on ways to prevent lawsuits from being filed in the first place. Recent studies and publications indicate that physicians may have more control over the lawsuit lottery than they realize. An article that appeared on the front page of the May 18, 2004 edition of the Wall Street Journal is reprinted below with permission. This article supports the proposition that the best tool to minimize the possibility of being sued may be as simple as expressing condolence and empathy when there is a bad outcome. The lawsuit reform bill that recently passed the Oklahoma legislature, H.B. 2661, contains an "I'm Sorry Law" that permits physicians to express condolence without those statements being used against them in court. For more information regarding the power of an apology, physicians may want to obtain the book by Michael S. Woods, M.D. (a speaker at the OSMA Physician Survival Summit) titled: "Healing Words: The Power of Apology in Medicine." The book can be obtained from: Doctors in Touch, 708.697.6447 or info@doctorsintouch.com. PMID- 15346805 TI - The economic impacts of Oklahoma's Family Medicine residency programs. AB - The enactment of Medicare and Medicaid created a new demand for medical services in Oklahoma, particularly in rural areas. The state of Oklahoma responded by creating The Oklahoma Physician Manpower Training Commission in 1975. The overall purpose of the Commission was to increase the number of primary care physicians and influence distribution into non-metro areas. This analysis concerns the public policy value of this ongoing program. The PMTC has provided resident stipend funding to each of Oklahoma's publicly funded Family Medicine residency programs. Since 1975, the PMTC has provided over 139 million dollars in resident stipend funding and support; and there have been 749 program graduates with 431 practicing in Oklahoma. This model calculates that the Oklahoma-based physicians have created a cumulative 3.7 billion dollars of economic impact on the state; and conservatively estimates that only 10% of the practice decisions/locations were influenced by the PMTC. This creates an estimated return of 370 million dollars on an "investment" of 139 million dollars. Additionally the model demonstrates that the current cohort of physicians is annually responsible for 15,530 jobs and an associated payroll of 428 million dollars. PMID- 15346806 TI - Rungs on the ladder: developing a career structure for European researchers. AB - When you are at the bottom of the career ladder in the life sciences it's often hard to see how high you can go or where it might lead you. In Europe, the academic ladder is missing at least one essential rung, and young researchers need better training to step out in new directions. PMID- 15346807 TI - The exosome, a molecular machine for controlled RNA degradation in both nucleus and cytoplasm. AB - One of the most important protein complexes involved in maintaining correct RNA levels in eukaryotic cells is the exosome, a complex consisting almost exclusively of exoribonucleolytic proteins. Since the identification of the exosome complex, seven years ago, much progress has been made in the characterization of its composition, structure and function in a variety of organisms. Although the exosome seems to accumulate in the nucleolus, it has been clearly established that it is also localized in cytoplasm and nucleoplasm. In accordance with its widespread intracellular distribution, the exosome has been implicated in a variety of RNA processing and degradation processes. Nevertheless, many questions still remain unanswered. What are the factors that regulate the activity of the exosome? How and where is the complex assembled? What are the differences in the composition of the nuclear and cytoplasmic exosome? What is the detailed structure of exosome subunits? What are the mechanisms by which the exosome is recruited to substrate RNAs? Here, we summarize the current knowledge on the composition and architecture of this complex, explain its role in both the production and degradation of various types of RNA molecules and discuss the implications of recent research developments that shed some light on the questions above and the mechanisms that are controlling the exosome. PMID- 15346808 TI - Regulation of nuclear functions--nucleocytoplasmic transport in context. PMID- 15346809 TI - The molecular composition of the sarcomeric M-band correlates with muscle fiber type. AB - The M-band is the transverse structure that cross-links the thick filaments in the center and provides a perfect alignment of the A-band in the activated sarcomere. The molecular composition of the M-bands in adult mouse skeletal muscle is fiber-type dependent. All M-bands in fast fibers contain M-protein while M-bands in slow fibers contain a significant proportion of the EH-myomesin isoform, previously detected only in embryonic heart muscle. This fiber-type specificity develops during the first postnatal weeks. However, the ratio between the amounts of myosin and of myomesin, taken as sum of both isoforms, remains nearly constant in all studied muscles. Ultrastructural analysis demonstrates that some of the soleus fibers show a diffuse appearance of the M-band, resembling the situation in the embryonic heart. A model is proposed to explain the functional consequence of differential M-band composition for the physiological and morphological properties of sarcomeres in different muscle types. PMID- 15346810 TI - Characterization of proteins secreted during maize microspore culture: arabinogalactan proteins (AGPs) stimulate embryo development. AB - To study molecules secreted from cultured plant cells that promote development, maize microspores were transferred into culture and the conditioned media were collected over time and analysed. Electrophoresis indicated that both non glycosylated and glycosylated proteins including arabinogalactan proteins (AGPs) appeared in the medium and their concentration increased during the time of culture. The development of embryos was correlated with the presence of specific extracellular proteins, using an experimental system based on a tunicamycin inhibition test. In addition, a precise protein analysis was conducted using MALDI-TOF and ESI-MS-MS techniques. These approaches have allowed the identification of 5 other types of proteins: a cell wall invertase, two thaumatin isoforms, one 1-3 beta-glucanase and two chitinase isoforms. Altogether these experiments and results open ways for research aimed at understanding which molecules stimulate embryo formation. Moreover, AGPs may be used to stimulate the development of microspores (pollen embryogenesis) prepared from non-responsive genotypes. PMID- 15346811 TI - Changes in pectins and MAPKs related to cell development during early microspore embryogenesis in Quercus suber L. AB - The occurrence and significance of changes in cell wall components and signalling molecules has been investigated during early microspore embryogenesis in cork oak (Quercus suber L.) in relation to cell proliferation and cell differentiation. Microspore embryogenesis has been induced in in vitro anther cultures of Q. suber by the application of a stress treatment of 33 degrees C. After the treatment, microspores at the responsive developmental stage of vacuolate microspore switched towards proliferation and the embryogenesis pathway to further produce haploid plantlets. Ultrastructural and immunocytochemical analysis revealed changes in cell organisation after induction at different developmental stages, the cellular features displayed being in relation to the activation of proliferative activity and the beginning of differentiation in young and late proembryos. Immunogold labelling with JIM5 and JIM7 antibodies showed a different presence of pectin and level of its esterification in cell walls at different developmental stages. Non-esterified pectins were found in higher proportions in cells of late proembryos, suggesting that pectin de-esterification could be related to the beginning of differentiation. The presence and subcellular distribution of Erk 1/2 MAPK homologues have been investigated by immunoblotting, immunofluorescence and immunogold labelling. The results showed an increase in the expression of these proteins with a high presence in the nucleus, during early microspore proembryos development. The reported changes during early microspore embryogenesis are modulated in relation to proliferation and differentiation events. These findings provided new evidences for a role of MAPK signalling pathways in early microspore embryogenesis, specifically in proliferation, and would confer information for the cell fate and the direction of the cell development. PMID- 15346812 TI - 'Fetal programming' and 'functional teratogenesis': on epigenetic mechanisms and prevention of perinatally acquired lasting health risks. AB - Alterations of the intrauterine and early postnatal nutritional, metabolic, and hormonal environment may cause predispositions to the development of disorders and diseases in later life. Mechanisms responsible for this perinatally acquired 'malprogramming' still remain unclear. It has long been known, however, that hormones are environment-dependent organizers of the developing 'neuroendocrine immune network', which regulates all fundamental processes of life. When present in nonphysiological concentrations during critical ontogenetic periods, hormones can therefore also act as 'endogenous functional teratogens'. Fetal and neonatal hyperinsulinism is a pathognomic feature in the offspring of diabetic mothers. Perinatal hyperinsulinism also occurs due to early postnatal overfeeding. Data obtained by our group indicate that elevated insulin concentrations during critical periods of perinatal life may induce a lasting 'malprogramming' of neuroendocrine systems regulating body weight, food intake, and metabolism. Similar characteristics may occur due to perinatal hyperleptinism, hypercortisolism etc. Since mechanisms of early 'programming' of obesity, diabetes, and the metabolic syndrome X are unclear, a complex 'neuroendocrine malprogramming' of the regulation of body weight and metabolism may provide a general etiopathogenetic concept in this context, exemplarily revealing critical new implications for chances and challenges of perinatal preventive medicine in the future. PMID- 15346813 TI - The baby or the bathwater: which one should be discarded? PMID- 15346814 TI - Waterbirths compared with landbirths: an observational study of nine years. AB - AIMS: This study compares neonatal and maternal morbidity and mortality between waterbirths and landbirths (spontaneous singleton births in cephalic presentation, vacuum extractions are excluded). METHODS: In this observational study covering nine years, standardized questionnaires were used to document 9,518 spontaneous singleton cephalic presentation births, of which 3,617 were waterbirths and 5,901 landbirths. RESULTS: Landbirths show higher rates of episiotomies as well as third and fourth degree perineal lacerations. Waterbirths show a higher rate of births "without injuries", first and second-degree perineal lacerations, vaginal and labial tears. After a waterbirth, there is an average loss of 5.26 g/l blood; this is significantly less than landbirths where there is an 8.08 g/l blood loss on average. In 69.7% waterbirths required no analgesic, compared to 58.0% for landbirths. Water and landbirths do not differ with respect to maternal and neonatal infections. After landbirths, there was a higher rate of newborn complications with subsequent transfer to an external NICU. During the study, there were neither maternal nor neonatal deaths related to spontaneous labor. CONCLUSIONS: Waterbirths are associated with low risks for both mother and child when obstetrical guidelines are followed. PMID- 15346815 TI - Anti-hypertensive therapy and the feto-placental circulation: effects on umbilical artery resistance. AB - OBJECTIVE: To investigate and compare the direct effects of compounds used in the treatment of hypertensive disease in pregnancy on human umbilical artery resistance in vitro. METHODS: Isometric tension recordings were performed under physiological conditions on human umbilical arterial rings (n=30). The in vitro effects of labetolol, hydralazine, alpha-methyldopa, nifedepine and magnesium sulphate (at concentration ranges from 1 nanomolar to 1 millimolar), and their respective vehicle controls, were measured. Results were expressed as -logEC50 (pD2) and mean maximal inhibition values for each compound. RESULTS: All compounds investigated, except alpha methyldopa, exerted a significant relaxant effect on umbilical arterial tone. Alpha-methyldopa was significantly less potent when compared to all other compounds (mean maximal inhibition value [20.89+/ 7.99%] versus all other agents [range 63.15+/-8.70-84.12+/-3.84%] (P<0.01)). The dose response curve of nifedipine yielded a significantly greater PD2 value when compared to that of hydralazine, labetalol, and magnesium sulphate (PD2 value [5.82+/-0.34] versus the above groups [range 3.10+/-0.09-3.52+/-0.14] (P <0.01)). CONCLUSION: These findings demonstrate that agents commonly used for the treatment of hypertensive disease in pregnancy, excluding alpha-methyldopa, have significant direct effects on the feto-placental circulation. These results suggest that alpha-methyldopa administration during pregnancy is less likely to produce significant direct effects on fetal vasculature then other agents used. PMID- 15346816 TI - A 2 hour versus the 3 hour 100 g glucose tolerance test for diagnosing gestational diabetes mellitus. AB - AIM: To determine how the omission of the third hour glucose measurement of the 100 g oral glucose tolerance test (GTT) affects the diagnosis of gestational diabetes mellitus (GDM). METHODS: Retrospective chart review of 876 women delivered in a tertiary care hospital in Israel during a three-year period, who underwent a 100 g 3 hour oral GTT following an abnormal 50 g glucose screen. GDM was diagnosed according to the "criterion standard" accepted in the Fourth International Workshop Conference on GDM. The results of the 100 g 3 hour oral GTT were then retrospectively re-evaluated by omission of the third hour plasma glucose measurement from the "criterion standard". RESULTS: GDM was diagnosed in 28.4% of the study. patients, while the omission of the third hour glucose measurement resulted in a 26.4% diagnosis of GDM. The perinatal data of the 18 omitted cases suggests that their exclusion from the GDM group would not have altered substantially the perinatal outcome of the study cohort. CONCLUSIONS: A 100 g 2 hour oral GTT is a simple and economic alternative to the 100 g 3 hour oral GTT. PMID- 15346817 TI - Mycoplasma hominis in mid-trimester amniotic fluid: relation to pregnancy outcome. AB - OBJECTIVE: The relationship between detection of Mycoplasma hominis in mid trimester amniotic fluid and subsequent pregnancy outcome was investigated. STUDY DESIGN: Amniotic fluids from 456 women of European background who underwent a transabdominal amniocentesis at weeks 15-17 of pregnancy were tested for M. hominis by polymerase chain reaction (PCR). The amplicons were hybridized to an internal probe and detected by ELISA. Pregnancy outcomes and clinical data were subsequently obtained. RESULTS: M. hominis were identified in 29 (6.4%) of the amniotic fluids. The rate of preterm labor in women positive for M. hominis (14.3%) was higher than in the negative women (3.3%) (p=0.01). Similarly, a spontaneous preterm birth with intact membranes occurred in 10.7% of the M. hominis-positive women as opposed to only 1.9% of the negative women (p = 0.02). The presence of this mycoplasma was not correlated with fetal chromosomal aberrations, intrauterine growth restriction or preeclampsia. CONCLUSIONS: Detection of M. hominis in second-trimester amniotic fluids can identify women at increased risk for subsequent preterm labor and delivery. PMID- 15346818 TI - Changes in the size of maternal inferior vena cava during pregnancy. AB - AIMS: To investigate the changes in the size of maternal inferior vena cava (IVC) during normal pregnancy. METHODS: The diameter and the area of IVC were measured in 32 non-pregnant and 214 pregnant women by ultrasonography. They were measured not only in the supine but also in the complete left lateral position when gestation was over 16 weeks. RESULTS: IVC in the 4-7 weeks pregnant group was larger than that in the non-pregnant group, and it decreased as gestation progressed during an early stage of pregnancy. After 16 weeks gestation, it demonstrated no change, at least for depending on the periods when the mother was in the supine position. On the contrary, they increased with the gestational progress as for the maternal left lateral position. CONCLUSIONS: The size of IVC at an early stage of pregnancy is enlarged, suggesting an underfilling state of arterial circulation. In the second and third trimesters, it is stationary when the mother is supine. In the left lateral position, it increases as the gestation progresses, reflecting blood volume expansion during pregnancy. PMID- 15346819 TI - Maternal plasma procalcitonin concentrations in patients with preterm labor and intact membranes--prediction of preterm delivery and admission-to-delivery interval. AB - The purpose of this study was to evaluate procalcitonin (PCT) plasma levels in pregnancy complicated by preterm labor and to determine their value in the prediction of preterm delivery and the length of the admission-to-delivery interval. The study population consisted of 53 patients with preterm labor and 31 healthy pregnants. The study patients were divided according to the delivery time and to the admission-to-delivery interval. Plasma PCT concentrations were higher in preterm labor than in healthy pregnants. Although at the onset of preterm labor plasma PCT concentrations in patients who delivered prematurely were higher than in patients who, after tocolytic treatment, delivered at term, the difference was not significant. Also in cases of preterm labor delivered within and after three and seven days of admission no differences were observed. The highest values in the prediction of preterm delivery and the length of admission to-delivery interval corresponded to a PCT concentration of 1.7 ng/ml. These findings suggest that although preterm labor is associated with increased PCT concentrations in maternal plasma, there is no significant association either between plasma concentration of PCT at the moment of threat and preterm delivery, or the admission-to-delivery interval. The predictive value of plasma PCT determinations is unsatisfactory. PMID- 15346820 TI - Determinant factors in Ecuador related to pregnancy among adolescents aged 15 or less. AB - OBJECTIVE: Determine social factors related to pregnancy among young adolescents. METHODS: We obtained socio-demographic data from early adolescent nulliparas aged < or = 15 years. RESULTS: During the study period at the Enrique C. Sotomayor Obstetrics and Gynecology Hospital, Guayaquil-Ecuador 201 early adolescent nulliparous patients of low socio-economic status were surveyed and compared with 201 low socio-economic nulliparous controls aged 20 to 30. Mean age of adolescents was 14+/-0.6 years, 58.7% of them were aged 14. Age of menarche and sexual initiation was lower among adolescents (11.8+/-0.9 vs 12.8+/-2 years and 12.6+/-1.1 vs 16.2+/-5 years, respectively, p<0.05). A higher rate of adolescents initiated sexually before menarche onset (18.4% vs 5%, p<0.05). The rate of those who wanted to become pregnant, had adequate prenatal care and had knowledge of conception, used contraception or had knowledge of any contraceptive method prior to pregnancy was significantly lower in adolescents (16.9% vs 86.5%; 37.3% vs 94.5%; 18% vs 70.1%; 6.5% vs 89.5%; 42.8% vs 84.5%, respectively, p<0.05). Age of sexual partner was lower in the adolescent group (20.4+/-3.4 vs 30+/-8 years, p<0.05). Compared to controls, higher rates of adolescents were school dropouts, had problems with the law and were involved in domestic violence (87% vs 9.9%, 7.5% vs 0.5%, 44.7% vs 2.5%, respectively, p <0.05). None of the adolescents were living with mother and father in a complete family structure. Parental illiteracy was higher among adolescents (9% vs 3.5%, p < 0.05). CONCLUSION: In this low socio-economic population, early sexual initiation, poor reproductive health knowledge and the disruption of family structure were the main social factors related to pregnancy among adolescents aged 15 or less. PMID- 15346821 TI - Different maternal serum hCG levels in pregnant women with female and male fetuses: does fetal hypophyseal--adrenal--gonadal axis play a role? AB - Fetal gender has a significant effect on maternal and cord blood hCG levels, particularly during the last trimester of the pregnancy. However, the reason for this difference is obscure. The aim of the present study was to investigate whether term fetal hypophyseal - adrenal - gonadal axis differs between female and male fetuses thereby causing different hCG levels. The study consisted of 60 women with singleton pregnancies in the third trimester. Thirty-one pregnant women were carrying female fetuses, whereas 29 were carrying male. Human chorionic gonadotropin (hCG), estradiol, progesterone, testosterone, dehydro epiandrosteron-sulfate (DHEAS), prolactin and growth hormone levels were measured in maternal serum and umbilical cord blood. In female bearing pregnancies maternal and cord blood hCG levels were significantly higher than in male bearing pregnancies (P<0.001). Maternal and cord blood estradiol, progesterone, testosterone, DHEAS, prolactin and growth hormone levels were not significantly different in either fetal gender. When all patients were considered as a group there were no correlations between fetal hCG levels and any of the measured hormones. Term fetal DHEAS, estrogen, progesterone, testosterone, growth hormone and prolactin levels do not contribute to different hCG levels between female and male fetuses. It is possible that fetal hypophyseal-adrenal gonadal axis does not play a central role as the cause of different hCG levels. PMID- 15346822 TI - Behavioral pattern continuity from prenatal to postnatal life--a study by four dimensional (4D) ultrasonography. AB - AIM: To investigate whether the same behavioral patterns were present pre- and postnatally, and whether there were any differences in the frequency of movements observed in fetal and in early neonatal life. SUBJECTS AND METHODS: Ten out of 37 pregnant women in the third trimester of pregnancy (median gestational age 34 weeks, range 33 to 35 weeks) in the two-month period (from November 1st to December 31st, 2003) were enrolled in the investigation. Ten term, appropriate for gestational age newborns (seven born vaginally, three by elective SC, six girls, six first-born) and were enrolled in the study. All 4D examinations were performed on Voluson 730 (Kretztechnik, Zipt, Austria) and Acuvix (Medison, Korea) with transabdominal 5 MHz transducer. After standard assessment in 2D B mode ultrasound, a 4D mode was switched on and live 3D image was reconstructed by selecting the ideal representative 2D image placed in the region of interest (ROI). The recordings of neonatal behavior were made on the Sony P-612 OHMPL videotape by video camera (Sony Camcoder CC DTRV 318 Hv8) and reviewed on the videocassette recorder (Sony VHS SLV-N 900). The median of newborns' age at the moment of recording was 49 hours (range 4 to 112). During the examination, newborns were lying in the bed, separated from other infants in the nursery, dressed, and lying on their backs in a supine position with unrestrained hands. The temperature in the room was 22 to 24 degrees C. The video recording was performed mainly while the children were actively awake or during alert inactivity. RESULTS: There were no movements observed in fetal life that were not present in neonatal life, while the Moro reflex was present only in neonates. The most frequent fetal and neonatal movements were scowling, eye and mouth opening, and hand to face, hand to eye and hand to head movements. Isolated blinking, mouth to eyelid movement, yawning, tongue expulsion and scowling were more frequent in neonates than in fetuses, although the difference was not statistically significant. Hand to mouth movements were more frequent in neonatal than in fetal life while all other hand movements were less frequent in neonates than in fetuses, although the differences did not reach statistical significance. Spearman rank order correlation reached statistical significance in smiling (R=0.71; t=2.91; P=0.02) and in hand to ear movement (R = 0.80; t= 3.86; P = 0.005), and was almost statistically significant in isolated eye blinking (R=0.61; t=2.17; P =0.06), while the correlations between the rest of the movements were not statistically significant. CONCLUSIONS: 4D ultrasonography is a powerful tool in the assessment of fetal behavior, and our study showed that there is a continuity from fetal to neonatal behavior, especially in terms of isolated eye blinking movements, mouth and eyelid opening, yawning, tongue expulsion, smiling, scowling and hand movements directed to other parts of the face. PMID- 15346823 TI - Evidence of fetal pulmonary aspiration of intra-amniotic administered surfactant in animal experiment. AB - INTRODUCTION: The instillation of surfactant into the airways of patients with respiratory distress syndrome (RDS), especially in the neonatal period, is a proven therapy. The preventive therapy of RDS through intra-amniotic injection of surfactant has been reported recently. It has not been conclusively shown, however, that the surfactant administered in this way actually reaches the fetal pulmonary airways. OBJECTIVE: To study the distribution in fetal organs of a natural surfactant labeled with technetium-99m and injected through amniocentesis into the amniotic sac of guinea pigs in the last third of pregnancy. METHODS: After stimulating fetal respiratory movements with aminophylline 0.3 ml of an aqueous suspension containing 0.75 mg of phospholipids of a natural bovine surfactant labeled with technetium-99m, together with 0.1 ml of the biological dye carmine indigo, were injected into the amniotic sac. One hour later fetuses were delivered by cesarean section. In those that were dye-stained, dosimetric and gammagraphic tests were applied to trachea, lungs, esophagus, stomach, heart, liver, kidneys and placenta. RESULTS: Significant radio isotopic activity was found in both lungs of six treated fetuses, with a dose capture of between 1.0% and 5.3% of total dose. The level of activity in the stomachs was similar to that in the lungs (0.9% to 3.0% dose capture), whereas activity in other organs was negligible except in two placentae. No radio isotopic activity was found in non injected control fetuses. CONCLUSIONS: In the present animal model natural surfactant injected intra-amniotically is aspirated into the lungs within one hour. PMID- 15346824 TI - Impact of phototherapy on vasoactive mediators: NO and VEGF in the newborn. AB - AIM: The objective of this study was to evaluate the effects of close and remote phototherapy on serum nitric oxide (NO) and vascular endothelial growth factor (VEGF) levels as well as on body temperature heart rate and blood pressure in neonates of different gestational ages. PATIENTS AND METHOD: Term (gestational age > or = 37 weeks) and preterm neonates (GA < 37 weeks) with hyperbilirubinemia requiring phototherapy were included in the study. All patients except for the ones in incubators were randomized to receive either close phototherapy (15 cm above the patient) or remote phototherapy (30-45 cm above patient). Body temperature, heart rate and blood pressure were measured before treatment, six hours into treatment and one hour after cessation of treatment. Blood samples for NO and VEGF measurements were also taken at the same times. RESULTS: Sixty-one term newborns and 37 preterm newborns were included in the study. Patients were distributed into four groups according to the dose of treatment together with gestational age, i.e. term close and remote photoherapy groups (n = 29, n = 32, respectively), preterm close and remote photoherapy groups (n=10, n=27, respectively). Body temperature increased significantly with phototherapy in all groups but was not at hyperthermia level. Heart rate increased in all groups except for term newborns in the remote phototherapy group and blood pressure decreased in term infants but was unchanged in preterms. None of these changes were at the level of tachycardia or hypotension for a newborn. Phototherapy did not result in elevation of NO or VEGF levels. CONCLUSION: This study showed that in our group of patients close or remote phototherapy caused some body temperature, heart rate and blood pressure changes that were not clinically significant and did not result in increased levels of NO or VEGF, which are well known vasodilator mediators. PMID- 15346825 TI - Reexpansion pulmonary edema following patent ductus arteriosus ligation in a preterm infant. AB - Reexpansion pulmonary edema (RPE) is rare and usually follows rapid reexpansion of a collapsed lung. We report on a preterm infant who developed pulmonary edema within an hour of surgical ligation of patent ductus arteriosus (PDA). There were no other cardiac anomalies, fluid overload or airway obstruction to explain the change in clinical status. With supportive treatment the patient's condition became stable and was extubated within 48 hours. Lung retraction for better field exposure is often needed when performing PDA ligation in preterm infants. Reinflation of a retracted lung is thought to be the cause of our patient's pulmonary edema. We conclude that RPE, although uncommon, may occur following surgical ductal ligation and that clinicians should be aware of such a possible complication. PMID- 15346826 TI - Prostaglandin E1 treatment in patent ductus arteriosus dependent congenital heart defects. AB - Prostaglandin E1 (PGE1) treatment can be life saving in patients suffering from ductus dependent congenital heart defect. We analyzed the indications and side effects of PGE1 therapy over a five-year period. The purpose of the study was also to examine whether a change in serum electrolyte levels could be detected. Forty-nine patients were treated with PGE1 during this period. PGE1 treatment was indicated by ductus dependent systemic circulation in 16 cases, ductus dependent pulmonary circulation in 17 cases, transposition of the great arteries in 13 cases and pulmonary hypertension (persistent fetal circulation) in three cases. As early side-effects of the treatment, fever occurred in 27/49 cases while apnoea was observed in 15 patients. In a one-week-old neonate with coarctation of the aorta grade III intraventricular hemorrhage developed. A mild decrease of sodium, potassium and chloride levels and a slight shift of pH levels toward metabolic alkalosis could be detected after one day and one week of PGE1 treatment. Because of these side-effects of PGE1 patients should be monitored in an intensive care unit. According to our observations electrolyte levels may exhibit a slight decrease; however, in the case of a short-term therapy extra salt supplementation is not necessary. PMID- 15346827 TI - Maternal anaphylaxis and fetal brain damage after intrapartum chemoprophylaxis. AB - We report a case of maternal anaphylaxis following intrapartum chemoprophylaxis. The term fetus developed severe brain damage as a consequence of intrapartum asphyxia. The lesions resulted from maternal hypotension following anaphylaxis. We discuss the feto-maternal risks and the controversial treatment of such a condition. The increasing number of penicillin-treated parturients will result in further cases of maternal anaphylaxis than previously found. PMID- 15346828 TI - Atypical sonographic presentation of fetal unilateral inguinoscrotal hernia in a multiple gestation. AB - Ultrasonographic demonstration of bowel peristalsis within the fetal scrotum has been described as a pathognomonic sign of inguinoscrotal bowel herniation. We present the sonographic features and neonatal outcome of a fetus with a scrotal mass seen at 34 weeks of gestation in a twin pregnancy. This mass was diagnosed postnatally as a non-reducible inguinoscrotal hernia, in which bowel peristalsis had not been observed by real-time ultrasound in utero. PMID- 15346829 TI - A case report of polysplenia syndrome associated with genital tract duplication anomaly. AB - Polysplenia syndrome is a condition that persists as a defect of lateralisation, the embryonic process by which the site of body organs is determined. The most frequent manifestations of this syndrome, in addition to polysplenia are complex cardiac malformations, situs inversus, and bilobed lungs. Laterality defects have been known to be due to autosomal recessive inheritance. We report a unique case of polysplenia syndrome in association with genital tract duplication anomaly. PMID- 15346830 TI - Hyperphenylalaninemia in a premature infant with heterozygosity for phenylketonuria. AB - Hyperphenylalaninemia in preterm neonates with heterozygosity for phenylketonuria has previously not been described. We report on a very low birth weight infant, born at a gestational age of 27+5 weeks with a birth weight of 1080 g. Due to a positive family history prenatal diagnosis for phenylketonuria was performed, revealing heterozygosity for classic phenylketonuria. Yet the girl showed hyperphenylalaninemia with a maximum serum phenylalanine concentration of 515 micromol/l on the eighth day of life. Phenylalanine-restrictive parenteral and enteral nutrition was kept from the eighth until the 41st day of life. At term serum phenylalanine concentrations had normalized. We hypothesize that heterozygosity for phenylketonuria may be a risk factor for hyperphenylalaninemia in preterm born infants. Prematurity and the resulting immaturity of liver function with the genetically determined reduced activity of phenylalanine hydroxylase might have caused hyperphenylalaninemia in this girl. PMID- 15346831 TI - Burnout and personality in intensive care: an empirical study. AB - Previous research into the causes of burnout has mainly been concerned with external triggers, such as onerous work criteria or organizational or social influences. Factors such as individual reactions and personality have largely been ignored as a possible etiology of burnout. In preparation for a long-term study, this general cross-sectional study investigates the relationship between burnout and personality variables. Different personality variables that have a possible impact on burnout were determined in a number of prestudies. The data were gathered from 119 people working in intensive care units. The Maslach Burnout Inventory (MBI) was used as well as certain subscales of the following personality questionnaires: Eysenck Personality Inventory (EPI), Inventory of Aggressivity (IA), Trier Personality Questionnaire (TPQ), Scales of Control (SC), Locus of Control (LC), and the Logo-test (LOGO). The scales of mental health, respectively Psychoprotection, external locus of control, and neuroticism, were confirmed as being statistically relevant concerning burnout. The application and significance of this study for future burnout research are discussed. PMID- 15346832 TI - Internal auditing in hospitals. AB - The authors analyzed two national surveys to determine answers for two basic questions: How do the roles of internal auditors compare with those of their counterparts in other industries and to what extent over the past 6 years have the activities of internal auditors changed? Internal auditors in hospitals allocate their time primarily to financial/compliance and operational types of audits, as do their counterparts. The current trend is toward more operational types of audits. In the early years of employment, staff turnover in hospitals is significantly higher than in all combined industries, often leading to internal auditors' filling other positions in the organization. Hospital staff salaries are higher than are salaries in other industries combined. Staff composition continues to reflect the growing presence of women in the field. The majority of internal auditing directors believe that their salaries are fair, would recommend internal auditing as a career position, and are treated as valued consultants in the organization. PMID- 15346833 TI - Impact of practice arrangements on physicians' satisfaction. AB - This research examines the effect of practice arrangements on five dimensions of physicians' satisfaction (i.e., personal factors, resources, peer review, profession, and state regulations) and the moderating effect of job autonomy and decision making on this relationship. This research finds that physicians who work for HMOs and hospitals are more satisfied with job resources, regulatory climate, and their professions, compared with physicians who are self-employed (solo and group practices). Physicians who work for HMOs and hospitals have less autonomy and decision-making power, compared with self-employed physicians. Also, job autonomy partially moderates the relationship between organizational arrangement and physicians' satisfaction with job resources and satisfaction with the profession. Decision making does not moderate the organizational arrangement and physicians' satisfaction relationship. PMID- 15346834 TI - No good deed goes unpunished: the Provena covenant case. PMID- 15346835 TI - A brief review of worker disability with a call for a consumer perspective. PMID- 15346836 TI - Assessment of aortic stenosis severity. PMID- 15346837 TI - Ischemic heart disease: metabolic approaches to management. AB - The number of patients with coronary artery disease and its risk factors is increasing in Western nations. New treatments for these patients may soon include a class of agents known as the metabolic modulators. This group of agents consists of the partial fatty acid oxidation inhibitors trimetazidine and ranolazine, as well as dichloroacetate, which promotes carbohydrate utilization. Metabolic modulators also include the nutriceuticals L-carnitine and D-ribose. The available evidence regarding the benefits of each of these five agents is reviewed. PMID- 15346838 TI - Impact of echocardiography on indications for surgery in chronic mitral and aortic regurgitation. AB - Echocardiography is the key examination in the assessment of mitral and aortic regurgitation, as it is able to describe the etiology and mechanism of the disease and assess its severity and its repercussions on cardiac cavities and pulmonary pressures. Surgery now tends to be indicated at earlier stages, largely based on criteria drawn from echocardiography. In severe mitral regurgitation, surgery is not disputed in presence of left ventricular ejection fraction < 60% and/or end-systolic diameter > 45 mm. In a selected group of patients aged < 75 years, with a very high likelihood of successful repair and a low operative risk, surgery should be considered earlier (if ejection fraction is > 60% and end systolic diameter < 45 mm) in degenerative regurgitation such as flail leaflets. In aortic regurgitation, surgery should be considered in presence of an acceptable operative risk as soon as end-systolic diameter exceeds 25 mm/m2 (and/or end-diastolic diameter > 70 mm) or resting ejection fraction < 55%. Surgery should be performed rapidly in dystrophic aortic regurgitation, independent of its severity, if the diameter of the ascending aorta exceeds 50 or 55 mm, or if it increases rapidly during follow-up. Echocardiography is therefore at the center of the strategic discussion concerning the indication for and timing of surgery. PMID- 15346839 TI - Images in cardiology: The electrocardiogram that saved a life. PMID- 15346840 TI - Images in cardiology: A saphenous vein graft aneurysm fistula causing pulmonary artery dissection. PMID- 15346841 TI - Thrombus aspiration prior to primary angioplasty in acute myocardial infarction: estimation of rescued myocardial tissue by return of ST-segment elevation. AB - BACKGROUND: The goal of the study was to validate the clinical benefit of adjunctive thrombus aspiration prior to balloon or stent angioplasty in patients with acute myocardial infarction. HYPOTHESIS: Adjunctive thrombus aspiration reduces no-reflow by eliminating peripheral thromboembolism. METHODS: In all, 143 consecutive patients underwent primary angioplasty for acute myocardial infarction in our institution. We excluded patients with subacute stent thrombosis or bundle-branch pattern of the electrocardiogram (ECG) and divided the remaining 131 into two groups: 62 patients who underwent intervention without a thrombus aspiration device (Group 1) and 69 patients who were treated as soon as a specifically designed thrombus aspiration catheter became available at our institution (Group 2). The intention was to use thrombus aspiration in every suitable case with proximal occlusion of a major vessel. The reduction of the sum of ST-segment elevation following angioplasty as a marker of rescued myocardial tissue was compared between the two groups. RESULTS: Thrombus aspiration was technically feasible in 23 of 25 attempted cases. The reduction of the sum of ST segment elevation following intervention was 0.41 +/- 0.39 mV in Group 1 and 0.37 +/- 0.42 mV in Group 2 (p > 0.05). CONCLUSIONS: Thrombus aspiration may be helpful during primary angioplasty in selected cases but does not improve the general outcome. Other reasons for "no-reflow," apart from peripheral thromboembolism, seem to be responsible for the lack of clinical benefit. PMID- 15346842 TI - Association of genetic polymorphisms in the fibrinogen and platelet glycoprotein genes with unstable angina in Chinese patients. AB - BACKGROUND: Inherited predisposition has been associated with coronary artery disease (CAD) in the white population. HYPOTHESIS: The objective of this study was to investigate the association between the risk of unstable angina (UA) and genetic factors believed to be associated with an increased tendency toward thrombosis (the variable number of tandem repeats [VNTR] polymorphism of the platelet glycoprotein [GP] Ib alpha gene, Pl(A1/A2) of the platelet GP IIIa gene, 448G/A of the Bbeta fibrinogen gene and Thr312Ala of the Aalpha fibrinogen gene) in Chinese patients with UA. METHODS: We performed a case/control study evaluating 69 Chinese patients (43 men, 26 women) with UA and 69 control subjects without CAD, individually matched for age and gender. The restriction fragment length polymorphism (RFLP) method was used to determine the genetic polymorphisms. RESULTS: The frequencies of GP Ib alpha C/B genotype and Bbeta fibrinogen 448A allele were higher in patients with UA (46.4 vs. 30.4%, odds ratio [OR] 1.977, 95% confidence interval [CI] 0.98-3.97, p = 0.054, and 49.3 vs. 20.3%, OR 3.816, 95% CI 1.797-8.103, p = 0.000, respectively). Only four subjects (two cases, two controls) with GP IIIa Pl(A2) allele were found, and there was no association between Aalpha fibrinogen Thr312Ala polymorphism and UA. CONCLUSIONS: Chinese patients with UA had increased frequencies of GP Ib alpha C/B genotype and Bbeta fibrinogen 448A allele. These data suggest that some genetic factors may influence the development of UA. PMID- 15346843 TI - Effectiveness of enhanced external counterpulsation in patients with left main disease and angina. AB - BACKGROUND: Enhanced external counterpulsation (EECP) is a noninvasive device that uses three pairs of sequentially inflated pneumatic cuffs applied to the lower extremities and synchronized with the heart beat to provide diastolic augmentation, increase coronary blood pressure and flow, venous return and cardiac output, and decrease afterload. HYPOTHESIS: This study examines the safety and effectiveness of EECP therapy in patients with significant left main coronary artery disease (LMD). METHODS: In all, 2,861 patients enrolled in the International EECP Patient Registry (IEPR) were divided into three groups, those without LMD (n = 2,377), those with LMD and prior CABG (n = 431), and those with unbypassed LMD (n = 53). RESULTS: Patients with LMD, with or without prior CABG, were significantly more likely to have triple-vessel disease (98.1 and 88.7%, respectively) than patients without LMD (41.9%). Post-EECP, 74% without LMD, 75% with LMD with prior CABG, and 65% with unbypassed LMD improved their Canadian Cardiovascular Society (CCS) angina by at least one class (p = NS). There were no differences in the mean decrease in weekly angina episodes (7.1 vs. 8.0 vs. 7.6) and in the mean frequency of weekly nitroglycerin use (6.6 vs. 8.1 vs. 8.9). At 6 month follow-up, the CCS class improved further in all three groups, and there was a further reduction in mean weekly angina episodes (4.7 vs. 4.6 vs. 5.3) and nitroglycerin use (6.5 vs. 6.8 vs. 8.2). Kaplan-Meier life table analysis 8 months after starting EECP demonstrated a major cardiovascular event rate of 11.2% in patients without LMD, 15.6% in LMD with CABG, and 24.3% in LMD without prior CABG. Late mortality in unbypassed LMD was 13.2% (confidence interval [CI] 3.3-23.1) versus 4.8% (CI 2.7-7.1) in LMD with CABG, and 2.8% (CI 2.1-3.5) without LMD (p = 0.0039 by log-rank test). CONCLUSION: Enhanced external counterpulsation is equally effective in relieving angina in patients with or without LMD. However, the significantly increased late mortality in patients with LMD without prior CABG suggests that early revascularization should be considered in these patients. PMID- 15346844 TI - Depletion of antioxidants is associated with no-reflow phenomenon in acute myocardial infarction. AB - BACKGROUND: No-reflow phenomenon is observed in approximately one-third of patients after percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI), and is associated with poor functional and clinical outcomes. On the other hand, the formation of free radicals in vasculature exerts deleterious effects on coronary microcirculation. HYPOTHESIS: We hypothesized that redox state in coronary circulation may play a crucial role in no-reflow phenomenon in AMI. METHODS: Consecutive 26 patients with first AMI who underwent primary PCI < 24 h after onset were enrolled. Before PCI, blood samples were obtained from coronary sinus to measure plasma or serum antioxidative vitamins (vitamin C, vitamin E, and beta-carotene) and antioxidative enzymes (extracellular glutathione peroxidase [GPX], superoxide dismutase, and catalase). After PCI, the corrected Thrombolysis In Myocardial Infarction (TIMI) frame count (CTFC) was measured in the target vessel. Patients with TIMI < or = 2 flow despite an optimal PCI result were designated as no-reflow group (Group NR, n = 6) and the others as reflow group (Group R, n = 20). RESULTS: Levels of vitamin C, vitamin E, and GPX before PCI were significantly lower in Group NR than in Group R. The CTFC correlated inversely with levels of vitamin C, vitamin E, and GPX (p < 0.05). CONCLUSIONS: Depletion of antioxidants is associated with no reflow phenomenon in AMI. These findings strongly suggest that the redox state in coronary circulation plays an important role in the pathogenesis of no-reflow phenomenon. PMID- 15346845 TI - Serum carboxyl-terminal propeptide of procollagen type I in exercise-induced left ventricular hypertrophy. AB - BACKGROUND: Left ventricular hypertrophy (LVH) induced by exercise is considered to be a physiologic adaptive mechanism without fibrogenic hyperactivity, as occurs in pathologic hypertrophy. HYPOTHESIS: This study investigated serum markers of collagen synthesis and echo parameters of left ventricular diastolic function (LVdf) in 22 male athletes. METHODS: Twenty-two highly competitive male athletes (10 cyclists, 12 soccer players) were studied with full history, clinical examination, Doppler echocardiogram, and serum concentration of the carboxyl-terminal propeptide of collagen type I (PIP). They were divided into two groups: normal left ventricular mass (N) with left ventricular mass index (LVMI) < 125 g/m2 (14 athletes) and LVH with LVMI > 125 g/m2 (8 athletes). RESULTS: Age, body surface area, blood pressure, heart rate, and systolic function were not different between the groups. Serum concentration of PIP (N: 163 +/- 44.1 microg/l, LVH: 172.7 +/- 61.2 microg/l--NS) and LVdf (early to atrial peak mitral flow velocity ratio: [E/A] N: 1.77 +/- 0.47, LVH: 1.98 +/- 0.70--NS, and early to atrial peak mitral annulus velocity ratio: [Ea/Aa] N: 2.63 +/- 0.70, LVMI: 2.55 +/- 0.90 LV 1.61--NS) were similar in both groups. CONCLUSIONS: Normal serum concentration of PIP in athletes with LVH in association with normal LVdf indicates the possibility that in this type of physiologic hypertrophy there is mainly an increase of myocyte size without interstitial fibrosis. PMID- 15346846 TI - Therapeutic strategies, immediate and mid-term outcomes in non-ST-segment elevation acute coronary syndromes with respect to age: a single-center registry of 488 consecutive patients. AB - BACKGROUND: Elderly patients with non-ST-elevation acute coronary syndromes (NSTE ACS) may receive benefit from an early invasive strategy. However, aged patients often suffer from comorbidities that may contraindicate an invasive approach and affect prognosis adversely. The impact of comorbidities on an invasive approach to NSTE-ACS in the elderly has not been fully investigated. HYPOTHESIS: This study sought to examine the outcome of an unselected population of patients with NSTE-ACS stratified according to age and treatment approach. METHODS: The feasibility and efficacy of an invasive strategy for NSTE-ACS and the 6-month outcome were assessed in 253 unselected consecutive patients > or = 70 years (elderly) and compared with those of 235 unselected consecutive patients < 70 years. RESULTS: Angiography was not performed in 69 patients (86% > or = 70 years) because of contraindications. In the whole population, the 6-month event rate was significantly higher in elderly compared with younger patients (22 vs. 14%; odds ratio 1.8, 95% confidence interval 1.1-2.9; p < 0.02). This difference was driven by the high event rate observed in the elderly with contraindications to angiography (47 vs. 16% in the elderly treated invasively; p < 0.002). On the other hand, no significant difference was observed in the 6-month event rate between elderly and younger patients undergoing an invasive approach (16 vs. 13%; p = 0.36). Contraindications to angiography-namely, creatinine > or = 1.5 mg/dl and elevated troponin I at admission--were the only independent predictors of 6 month outcome. CONCLUSIONS: The invasive approach was feasible in 77% of patients > or = 70 years. Those with contraindications to angiography showed a poor mid term prognosis. The early invasive strategy was associated with more favorable outcomes regardless of age. PMID- 15346847 TI - Effect of quinapril on the attenuated heart rate recovery of type 2 diabetic subjects without known coronary artery disease. AB - BACKGROUND: Heart rate (HR) recovery at 1 min is a measure of the vagal reactivation that occurs after cessation of exercise. Despite ample evidence about the association of attenuated HR recovery with increased mortality, pharmacologic modification of this predictor has not been shown. On the other hand, angiotensin-converting enzyme (ACE) inhibitors are known to have vagomimetic activity. HYPOTHESIS: We hypothesized that ACE inhibition would increase HR recovery in a group of subjects known to have reduced HR recovery, namely diabetics. METHODS: Maximal treadmill exercise stress test was performed in 31 type 2 diabetic and 31 nondiabetic male subjects with similar age, body mass index, and hypertensive status. None of the subjects had known heart disease or evidence of myocardial ischemia during the test. The diabetic subjects, after 2 weeks of treatment with quinapril, underwent a second exercise test. A third test was performed 2 to 3 weeks after cessation of quinapril treatment. RESULTS: At baseline, despite similar exercise capacity, the diabetics had a lower HR recovery at 1 min than nondiabetics (25 +/- 8 vs. 31 +/- 8 beats/min, p < 0.01). Quinapril significantly increased HR recovery at 1 min in diabetics (25 +/- 8 beats/min at baseline vs. 28 +/- 8 beats/min with quinapril vs. 25 +/- 7 beats/min off-therapy, p < 0.01 by analysis of variance). CONCLUSIONS: The attenuated HR recovery of type 2 diabetics can be improved by quinapril. Whether the improvement in HR recovery with ACE inhibition can lead to decreased mortality is currently unknown. PMID- 15346848 TI - Heinrich Ewald Hering and the carotid sinus reflex. PMID- 15346850 TI - A method of obtaining starting values of k(in) and k(out) for the indirect response models. AB - A method based on the multivariate technique known as principal component analysis is proposed to obtain starting values for the rate constants of indirect response models. The method is not iterative and only requires standard deviation calculations for two quantities, which are simple functions of the measured pharmacodynamic response. An algorithm is provided which can be implemented in a spreadsheet. The methodology is justified theoretically herein, nevertheless, two examples are provided to illustrate the method and demonstrate its viability. PMID- 15346849 TI - Stereoselective hepatic disposition of model diastereomeric acyl glucuronides. AB - Numerous studies have previously been conducted with the impulse-response isolated perfused rat liver (IR-IPRL) to establish the role of both physiological and physicochemical factors in determining solutes' pattern of hepatic disposition, however the impact of optical isomerism on hepatic disposition has hardly been studied using this methodology. In this study, the IR-IPRL was used to assess the extent of stereoselectivity exhibited by the kinetic processes involved in the hepatic disposition of the diastereomeric acyl glucuronides of (R)- and (S)-2-phenylpropionic acid (i.e. (R)- and (S)-PPAG). Moment and model dependent (distributed model and axial dispersion model) analyses were conducted of the hepatic outflow profiles generated upon bolus administration of (R)-(14)C PPAG or (S)-(14)C-PPAG and 3H-Sucrose (used as a marker of the hepatic vascular space) into the portal inflow of isolated perfused livers of male Sprague-Dawley rats (n = 4). Significant differences between (R)- and (S)-PPAG were apparent in the pharmacokinetic parameters defining the total hepatic disposition of the two diastereomers, the most marked being the hepatic availabilities, where the value for (S)-PPAG (0.721 +/- 0.059) was significantly lower than that of (R)-PPAG (0.909 +/- 0.042). The distributed and axial dispersion model analyses suggested that the more extensive hepatic extraction of (S)-PPAG was (at least in part) due to the higher sinusoidal membrane permeability-surface area product (PS UPT) of the diastereomer, and this has been considered in light of the emerging evidence regarding the role of hepatocellular membrane transport mechanisms. Furthermore, given the potential immunogenicity of acyl glucuronides (through covalent binding to plasma and intracellular proteins), the results of this study suggest that diastereomeric glucuronides may exhibit differing toxicity due to differences in their access to intracellular proteins. PMID- 15346851 TI - A population pharmacokinetic analysis of milrinone in pediatric patients after cardiac surgery. AB - The purpose of this study was to ascertain the optimal pharmacokinetic model for milrinone in pediatric patients after cardiac surgery when milrinone was administered as a slow loading dose followed by a constant-rate infusion. The data used for pharmacokinetic analysis were collected in a prospective, randomized, placebo-controlled multi-center trial of milrinone as prophylaxis for the development of low cardiac output syndrome after surgery for repair of complex congenital cardiac defects. Two blood samples were randomly collected from each patient for determination of plasma milrinone concentrations with subsequent population pharmacokinetic modeling. The pharmacokinetics of milrinone in pediatric patients under 6 year's age were best described by a weight normalized one compartment model after a slow loading dose followed by a constant rate infusion. The volume of distribution was 482 ml kg(-1) and was independent of age. Clearance was a linear function of age given by Cl = 2.42 ml kg(-1) min( 1) [1 + 0.396*age]. PMID- 15346852 TI - Evaluation of type I error rates when modeling ordered categorical data in NONMEM. AB - The development of non-linear mixed pharmacokinetic/pharmacodynamic models for continuous variables is usually guided by graphical assessment of goodness of fit and statistical significance criteria. The latter is usually the likelihood ratio test (LR). When the variable to be modeled is categorical, on the other hand, the available graphical methods are less informative and/or more complicated to use and the modeler needs to rely more heavily on statistical significance assessment in the model development. The aim of this study was to evaluate the type I error rates, obtained from using the LR test, for inclusion of a false parameter in a non-linear mixed effects model for ordered categorical data when modeling with NONMEM. Data with four ordinal categories were simulated from a logistic model. Two nested multinomial models were fitted to the data, the model used for simulation and a model containing one additional parameter. The difference in fit (objective function value) between models was calculated. Three types of models were explored; (i) a model without interindividual variability (IIV) where the addition of a parameter describing IIV was assessed, (ii) a model with IIV where the addition of a drug effect parameter (either categorical or continuous drug exposure measure) was evaluated, and (iii) a model including IIV and drug effect where the inclusion of a random effects parameter on the drug effect was assessed. Alterations were made to the simulation conditions, for example, varying the number of individuals and the size and distribution of the IIV, to explore potential influences on the type I error rate. The estimated type I error rate for inclusion of a false random effect parameter in model (i) and (iii) were, as expected, lower than the nominal. When the additional parameter was a fixed effects parameter describing drug effect (model(II)) the estimated type I error rates were in agreement with the nominal. None of the different simulation conditions tried changed this pattern. Thus, the LR test seems appropriate for judging the statistical significance of fixed effects parameters when modeling categorical data with NONMEM. PMID- 15346853 TI - A Bayesian approach to tracking patients having changing pharmacokinetic parameters. AB - This paper considers the updating of Bayesian posterior densities for pharmacokinetic models associated with patients having changing parameter values. For estimation purposes it is proposed to use the Interacting Multiple Model (IMM) estimation algorithm, which is currently a popular algorithm in the aerospace community for tracking maneuvering targets. The IMM algorithm is described, and compared to the multiple model (MM) and Maximum A-Posteriori (MAP) Bayesian estimation methods, which are presently used for posterior updating when pharmacokinetic parameters do not change. Both the MM and MAP Bayesian estimation methods are used in their sequential forms, to facilitate tracking of changing parameters. Results indicate that the IMM algorithm is well suited for tracking time-varying pharmacokinetic parameters in acutely ill and unstable patients, incurring only about half of the integrated error compared to the sequential MM and MAP methods on the same example. PMID- 15346854 TI - A comparative study of seed crystals for the phosphorus crystallization process. AB - This study was performed to select a preferred seed crystal material for the phosphorus crystallization process through a comparative study of four materials: electron arc furnace, blast furnace and converter slag, and phosphate rock. Leaching and phosphorus removal tests were conducted to evaluate the efficacy of the four materials as seeding agents. Converter slag demonstrated a much larger leaching capacity with respect to calcium and hydroxide ions than did either electron arc furnace or blast furnace slag. The average phosphorus removal efficiencies of phosphorus rock, and the electron arc furnace, blast furnace and converter slag were 35.9%, 74.2%, 99.1%, and 94.5% following 20, 17, 32, and 175 days of leach time, respectively. Consideration of both technical and cost factors suggests that converter slag may have competitive advantages over the other three seed materials. X-ray diffraction (XRD) analysis showed that peak patterns for hydroxyapatite increased slightly as the hydrous flow time was extended. Scanning electron micrograph (SEM) images revealed that finely distributed cubic crystals were deposited on the surfaces of converter slag. When analyzed by energy dispersive x-ray spectrometer (EDS) mapping, the crystals gave a composition mole ratio (Ca/P) of 1.97 and they were determined to be hydroxyapatite. PMID- 15346855 TI - Bench-scale assessment of the effectiveness of an anaerobic selector in controlling filamentous bulking. AB - A laboratory scale sequencing batch reactor (SBR) was used to assess the ability of an anaerobic selector to control filamentous growth in an activated sludge plant experiencing bulking problems. The SBR was operated under a sequence of 60 min anaerobic followed by 120 min of aerobic period at 20 degrees C, with a total solids retention time (SRT) of 4 d and a hydraulic retention time (HRT) of 8 h. The apparent soluble organic carbon (SOC) removal from the liquid phase averaged 18% during the anaerobic period, which is significantly less than the 80-90% removal generally considered to be required to suppress the growth of filamentous bacteria. The anaerobic selector was, however, able to control sludge bulking, resulting in an average sludge volume index (SVI) below 140 ml g(-1). The true amount of the SOC uptake was masked by simultaneous hydrolysis of particulate organic matter during the anaerobic period, a process which should be considered in future studies and comparisons. PMID- 15346856 TI - Adsorption of phthalates by activated sludge and its biopolymers. AB - This study shows diethyl phthalate (DEP) and dibutyl phthalate (DBP) were substantially adsorbed by activated sludge and its extracellular polymeric substance (EPS). The adsorption characteristics followed Freundlich and Langmuir isotherms. According to the Langmuir isotherm, each gram of activated sludge at maximum adsorbed 0.73 mg of DEP and 17.6 mg of DBP, and each gram of centrifugation-extracted EPS adsorbed 14.3 mg of DEP and 10.6 mg of DBP. The adsorption increased with the hydrophobicity of phthalates. This suggests most phthalates, which are of higher hydrophobicity than DEP and DBP, are likely to be removed from wastewater through adsorption by the activated sludge in the biological treatment process. PMID- 15346857 TI - Application of the IAS theory combining to a three compartments description of natural organic matter to the adsorption of atrazine or diuron on activated carbon. AB - The study of natural organic matter (NOM) adsorption on an activated carbon showed that equilibrium cannot be described according to a simple model such as a Freundlich isotherm and confirms the need for a closer description of the organic matter to simulate the competitive adsorption with micropollutants. A representation of the organic matter in three fractions is chosen: non adsorbable, weak and strong adsorbable. The Ideal Adsorbed Solution Theory (IAST) can, under restrictive conditions, be used to effectively predict the competition between the pesticides and the organic matter. Therefore, it was noted that the model simulated with good precision the competition between atrazine or diuron and natural organic matter in aqueous solution for two activated carbons (A and B). The same parameters for the modeling of organic matter adsorption (Freudlich constants for two absorbable fractions) are used with the two pesticides. However, IAST does not allow correct modeling of pesticide adsorption onto two other (C and D) activated carbons in solution in natural water to be described. IAS theory does not reveal competition between diuron and NOM and pore blockage mechanism by the NOM is proposed as the major effect for the adsorption capacity reduction. However, the difference observed between the two pesticides could be due to in addition to the pore blockage effect, a particular phenomenon with the diuron, especially with D activated carbon. We can suppose specific interactions between the diuron and the adsorbed organic matter and a competition between adsorption sites of NOM and activated carbon surface. PMID- 15346858 TI - Impact of seasonal variation on treatment of swine wastewater. AB - Swine wastewater (agricultural wastewater) is normally stored in a storage holding tank for a certain time before released to the other treatment units, such as anaerobic lagoon and aerobic wetland. One of the characteristics for this treatment approach is that all the processes are open systems that are generally more passive in design and operation. As a consequence, seasonal variability including temperature and precipitation can have substantial impact on treatment efficacy and effluent water quality. This paper examines seasonal impacts of temperature on swine wastewater quality and treatment efficacy at a farm in East Leicester, Nova Scotia, Canada. During warm temperatures denitrification was noticeable in the anaerobic conditions, which would reduce the TSS removal rate from 76.6% in moderate temperatures to 42.1% in the warmest period recorded. Rainfall improved final effluent water quality, although this was shown to be through dilution rather than improvement of treatment efficacy. Following precipitation events the contaminant removals were negatively impacted in the aerobic lagoon, as BOD5 removal decreased from 61.6% before rainfall to 41.5% after rainfall, TSS from 71.4% to 59.3%, VSS from 73.4% to 59.3%, TKN from 59.9% to 42.1%, and NH4+ -N from 51.3% to 41.6%. In comparison to the aerobic conditions, the removal rates were increased for anaerobic condition with the rainfall dilution (e.g., TSS from 18.2% to 34.3%), which lead to an overall treatment improvement for the entire system. Thus the case study data presented in this paper provides an assessment of the operational and design issues that are particularly relevant for passive treatment systems that are used in the agriculture industry. PMID- 15346859 TI - Final decontamination of the filtrates obtained after interaction of two ligand containing Cu2+ and Zn2+ electroplating solutions. AB - A cheap and simple way to decontaminate two ligand-containing rinsing waters of metal finishing has been previously suggested. Both the copper diphosphate containing rinsing wastewater and acidic zinc ammonium-containing rinsing wastewater may be decontaminated by mixing them which results in precipitation of solid solutions of copper-zinc-potassium-ammonium diphosphates. This way of decontamination requires no expensive reagents since only a small amount of H2SO4, is needed for pH adjustment. 80-99.5 % of environmentally dangerous substances, viz. zinc, copper and diphosphate, are removed from the mixture. However, Cu2+ and Zn2+ amounts in filtrates significantly exceed the discharge consent level (DCL). Besides, high concentrations of undesirable diphosphate and ammonium ions, which cause the eutrophication of natural water reservoirs, are present in the filtrates. There are relatively small concentrations of heavy metals: Cu2+ (approximately 0.4 mM dm-3) and Zn2+ (approximately 1 mM dm-3), and relatively large quantities of ligands: P2O7(4-) (approximately 20 mM dm-3) and NH4+/NH3 (approximately 500 mM dm-3) in these filtrates. Our investigation has shown, that decontamination may be easily achieved in industry by using cheap materials or industrial by-products: lime, MgO, phosphogypsum, extractive phosphoric acid and spent steel etching solution. Using lime, Cu2+ and Zn2+ ions may be removed below their DCL. After that, diphosphate ion concentration may be reduced approximately 1000 fold by addition of spent steel etching solution or continuous mixing with phosphogypsum. The amount of NH4+/NH3 may be reduced 20 to 30 fold, the concentration of soluble phosphates -- 5 to 40 fold and both c Cu2+ and c Zn2+ -- lower than their DCL by precipitating barely soluble fine crystalline MgNH4PO46H2O (struvite). PMID- 15346860 TI - The removal of Direct Orange 39 by pulsed corona discharge from model wastewater. AB - Untreated wastewater from the dye industry and dyehouses cannot be directly discharged into the environment due to the high content of organic matter and intensive colouration, even with low concentrations of dye. In this paper, the application of a high voltage pulsed electrical discharge in the aqueous phase has been assessed for the dye degradation. Experiments were conducted in a batch reactor using model wastewater of the commercial water-soluble monoazo dye C.I. Direct Orange 39 (DO39). The effects of zeolite and ferrous sulphate in combination with the corona discharge were examined. Experiments were conducted for a range of process parameters including pH, conductivity, type and amount of zeolite, and ferrous sulphate concentration. A mathematical model to describe the kinetics of DO39 degradation in the corona reactor was developed. Aqueous phase pulsed streamer corona discharge as a method for coloured wastewater treatment showed very high effectiveness in the case of iron salt addition (Fenton's reaction). Low pH enhanced dye removal by corona in the absence of zeolite, thus implying that the acid properties of zeolites are important in dye degradation. Ecological parameters such as COD, TC, IC, TOC and IC50 measured before and after corona treatment showed that the treated wastewater can be discharged into the environment or reused as process water. PMID- 15346861 TI - Fe(III)-EDTA complex abatement using a catechol driven Fenton reaction combined with a biological treatment. AB - A combined chemical oxidation (catechol-driven Fenton reaction) followed by a biological treatment was used to degrade Fe(III)-EDTA (1.34 mM). The chemical treatment was inspired in fungal non-enzymatic wood rot mechanisms that use dihydroxybenzens in order to promote the Fenton reaction to breakdown wood structures. This chemical pre-oxidation used catechol (50 microM) and H202 (20 mM) and the reaction products were identified by GC-MS. In addition, a biological treatment was coupled using the baker's yeast Saccharomyces cerevisiae. The combined chemical biological treatment achieved 100% EDTA degradation, 68 % total organic carbon removal and 90% iron removal. PMID- 15346862 TI - Development and analysis of anaerobic biofilms onto hydrophobic and hydrophilic surfaces. AB - Fluorescent in situ hybridization (FISH) with domain and group specific probes that target intracellular 16S rRNA were used to investigate microbial composition of anaerobic biofilms developed on polypropylene (hydrophobic) and glass (hydrophilic) surfaces fitted inside a Modified Robbins Device (MRD). Crushed anaerobic granular sludge was used as inoculum for biofilm development in the MRD. The inoculum and biofilms formed showed nearly the same microbial composition, both were dominated by hydrogenotrophic methanogenic Archaea related to the Methanobacteriaceae as detected by the specific probe (MB1174). This group accounted for 44 to 90% of the DAPI-stained cells. Cells which hybridized to the Bacteria specific probe (EUB338) accounted for 3-18% of the DAPI-stained cells. After the first day of the biofilm formation experiment, a larger number of cells, 4.6 x 10(4) cells mm-2, could be seen colonizing the polypropylene coupon compared to the glass, 8.2 x 10(3) cells mm-2. However, after 9 days these numbers were very similar, i.e. 6.3 x 10(5) cells mm-2 and 7.2 x 10(5) cells mm 2, for the glass and polypropylene coupons, respectively. Our data suggest that the hydrophobicity of the support material did not influence the initial development and the microbial composition of anaerobic biofilms developed in the MRD. PMID- 15346863 TI - Evaluation of the sorption properties of a Mexican organo clinoptilolite-rich tuff for phenol and 4-chlorophenol. AB - A Mexican clinoptilolite-rich tuff was modified with the hexadecyltrimethyl ammonium (HDTMA) and used for the sorption of phenol and 4-chlorophenol. Whereas the zeolitic tuff had no affinity for phenol and 4-chlorophenol, the modified zeolite removed these organic compounds. The sorption behavior from aqueous solutions was studied by means of batch type and column experiments and isotherms were determined. It was found that the uptake was higher for 4-chlorophenol than for phenol. PMID- 15346864 TI - Fate of E. coli across mechanical dewatering processes. AB - Five UK sludge treatment plants have been monitored for Escherichia coli (E.coli) variation after mechanical belt press and centrifuge dewatering processes. A complementary laboratory trial was also completed to examine the effects of varying centrifugal force on raw sludge E.coli content. An E.coli balance between the numbers contained in the flows entering and exiting four full scale centrifuge dewatering systems indicated a minimum 63 % increase in E.coli numbers between the input feed and sludge cake for a digested sludge input to the centrifuge. For two of the centrifuge sites this increase was statistically significant and corresponded to an increase in E.coli concentration ranging up to 1.4 Log after centrifugation. However, E.coli variation was found to be dependent on the type of sludge, as centrifuge dewatering of raw sludge at full scale resulted in a 40 % decrease in E.coli numbers. The complementary laboratory centrifuge work confirmed that E.coli numbers decreased in raw sludge after centrifugation. E.coli numbers were not observed to increase in digested sludge which had been dewatered using a belt press. A decrease of 44 % was observed. PMID- 15346865 TI - Growth of water hyacinth in municipal landfill leachate with different pH. AB - Batch experiments were conducted to investigate the effect of municipal landfill leachate pH on the growth of water hyacinth (Eichhornia crassipes). These experiments were carried out in a green house environment on leachate samples collected from Essex-Windsor Regional Landfill, Windsor, Ontario, Canada. It was found that water hyacinth plants survived in a pH range of 4.0 to 8.0. Both alkaline pH (above 8.0) and highly acidic pH (below 4.0) had inhibitory effect on the growth of plants. The pH range, for optimum growth of the water hyacinth plants was found to be 5.8 to 6.0. At optimum growth, water hyacinth had an average mean relative growth rate of 0.043 d-1. It was found that nitrogen compounds underwent different transformations depending on the pH of leachate. Plant uptake, nitrification and volatilization were among these transformations. PMID- 15346866 TI - Sensitivity of embryotoxicity test with Mytilus galloprovincialis (Lmk) towards some compounds of environmental interest (copper and pesticides). AB - Some compounds of environmental interest were investigated using the toxicity test with early life stage of bivalve molluscs. Three pesticides (Carbofuran, Atrazine and Malathion), for which literature data are available only for some species of oysters, were tested with Mytilus galloprovincialis (Lmk). The toxicity of copper was investigated both with M. galloprovincialis and with Crassostrea gigas (Thunberg). Bioassays highlighted a good sensitivity towards the compounds, with EC50 values in the order of mg l(-1) for the pesticides and of microg l(-1) for copper. Comparison with other toxicity tests for aquatic environments showed that embryotoxicity test with bivalve is slightly more sensitive than embryotoxicity test with the sea urchin Paracentrotus lividus (Lmk), remarkably more sensitive than Microtox test but less sensitive than tests with the copepod Tigriopus brevicornis. PMID- 15346867 TI - Evaluation of soil gas sampling and analysis techniques at a former petrochemical plant site. AB - Methods for soil gas sampling and analysis are evaluated as part of a research study on soil vapour intrusion into buildings, conducted at a former petro chemical plant site ("Chatterton site"). The evaluation process was designed to provide information on reliability and selection of appropriate methods for soil gas sampling and analysis, and was based on a literature review of data and methods, and experiments completed as part of the research study. The broader context of this work is that soil gas characterization is increasingly being used for input into risk assessment of contaminated sites, particularly when evaluating the potential intrusion of soil vapour into buildings. There are only a limited number of research studies and protocols addressing soil gas sampling and analysis. There is significant variability in soil gas probe design and sample collection and analysis methods used by practitioners. The experimental studies conducted to evaluate soil gas methods address the permeation or leakage of gases from Tedlar bags, time-dependent sorption of volatile organic compound (VOC)-vapours onto probe surfaces and sampling devices, and analytical and quality control issues for light gas and VOC analyses. Through this work, common techniques for soil gas collection and analysis are described together with implications for data quality arising from the different methods used. Some of the potential pitfalls that can affect soil gas testing are identified, and recommendations and guidance for improved protocols are provided. PMID- 15346868 TI - Celiac disease. PMID- 15346869 TI - Decision time. PMID- 15346870 TI - Outcomes two, three and four years after comprehensive care under general anaesthesia. AB - OBJECTIVES: The objective was to review the outcomes for three groups of children after two, three and four years following comprehensive dental treatment under general anaesthesia (GA) for 292 children aged 1.8 to 5.9 years of age. DESIGN: The study was a retrospective review of ongoing dental treatment needs following treatment under GA. METHODS: Information including the individual tooth treatments carried out, ongoing care, fluoride status, socioeconomic status and ethnicity was obtained from dental records from the School of Dentistry and the Otago District Health Board School Dental Service. The accuracy of the records was evaluated by clinically examining 10 percent of the children. RESULTS: Ninety five percent of the treated children were followed up. Fifty-five percent had new caries recorded. For the treatments provided, amalgam had a mean success of 57.1 percent, composite 73.4 percent, compomer 85.2 percent, stainless steel crowns 92.8 percent and pulpotomies 84.6 percent. The majority of replaced restorations were because of new carious lesions. CONCLUSIONS: This study indicated that most of the restorative procedures and materials used have very successful outcomes in these high-risk children. Ongoing risk of dental caries is high despite current preventive approaches. Other preventive approaches should be investigated. PMID- 15346871 TI - Prevalence of self-reported hand dermatoses in New Zealand dentists. AB - OBJECTIVES: To estimate the self-reported prevalence of hand dermatoses and irritation to the eyes, nose and airway among New Zealand dentists. MATERIALS AND METHODS: A cross-sectional survey used a postal questionnaire sent to all practising dentists on the New Zealand Dental Register. RESULTS: The response rate was 81.3 percent. Over 40 percent of dentists had experienced symptoms at some stage during their practising life, and one-third reported experience of symptoms during the previous 12 months. Prevalence was higher among females, more recent graduates, and those suffering from hay fever, asthma or any other allergic condition. One-third of sufferers had consulted a medical practitioner about the problem. Multivariate modelling showed that females had twice the odds of experiencing symptoms at some time during their practising life (after controlling for time since graduation, allergies and hay fever, lifelong wearing of gloves, and hobbies involving exposure to solvents). Those who reported hobbies involving solvents were over 11 times as likely to report having experienced symptoms at some time during their practising life. CONCLUSIONS: A substantial proportion of the dental practice workforce in New Zealand is at risk for occupational dermatoses. Reducing exposure to potential allergens and irritants is the key to minimising that risk. PMID- 15346872 TI - Dental erosion: part 2. The management of dental erosion. AB - Dental erosion is an important cause of tooth tissue loss in both children and adults. An earlier article (Mahoney and Kilpatrick, 2003) discussed the prevalence and causes of dental erosion. This second article will discuss the management of this condition which is often complicated by the multifactorial nature of tooth wear. Management of dental erosion can be considered in three phases: immediate, interim and long-term. Immediate management includes the early diagnosis of dental erosion, recording the status of the disorder at baseline and implementing appropriate preventive strategies including those aimed at reducing the acidic exposure as well as those that attempt to increase an individual's resistance to erosive tooth tissue loss. Interim and long-term treatment includes the provision of temporary diagnostic restorations, ongoing monitoring of disease progression, definitive restorative work where appropriate, and modification and reinforcement of preventive advice. Each phase will be discussed in light of current evidence. PMID- 15346873 TI - Reflections on a career move. PMID- 15346874 TI - One hundred years on. PMID- 15346875 TI - The Down syndrome patient in dental practice, Part II: clinical considerations. AB - Down syndrome (DS), or Trisomy 21, is a genetic disorder which results in intellectual impairment, typical craniofacial features and a wide spectrum of phenotypic abnormalities (reviewed by Kieser et al, 2003). Despite a large volume of research into this condition, its prevalence remains high (1 in 700 live births - Serra and Neri, 1990; Reeves, 2000). This, together with a notable increase in life expectancy (Pilcher, 1998), improved health care and a greater degree of integration into society in New Zealand has resulted in an increased demand for dental care. PMID- 15346876 TI - Water fluoridation and dental caries in 5- and 12-year-old children from Canterbury and Wellington. AB - OBJECTIVES: Claims have been made that the effectiveness of water fluoridation has reduced due to the widespread availability of other sources of fluoride. This study examines the differences in the oral health of children living in fluoridated and non-fluoridated areas of Canterbury and Wellington, New Zealand. DESIGN: The data used in this cross-sectional study had been routinely collected into a computerized data-collection system by the School Dental Services in the two study areas. SUBJECTS AND METHODS: Records of dental status (dmfs/DMFS), fluoridation status, ethnicity, and socio-economic status for 8030 5-year-olds, and 6916 12-year-olds in 1996 were analysed. RESULTS: Caries prevalence and severity was consistently lower for children in the fluoridated area for both age groups, and within all subgroups. Five-year-olds in the fluoridated area had 2.63 dmfs (sd, 5.88), and those in the non-fluoridated area 3.80 dmfs (sd, 6.79). For 12-year-olds the respective figures were 1.39 DMFS (sd, 2.30) and 2.37 DMFS (sd, 3.46). Multivariable analysis confirmed the independent association between water fluoridation and better dental health. CONCLUSIONS: This results of this study show children living in a fluoridated area to have significantly better oral health compared to those not in a fluoridated area. These differences are greater for Maori and Pacific children and children of low socio-economic status. PMID- 15346877 TI - The involvement of New Zealand dentists in smoking cessation interventions. AB - OBJECTIVES: To determine the attitudes and practices of dentists towards smoking cessation interventions (SCI), to identify perceived barriers to providing SCI, and to determine the level of interest in future training in smoking cessation for dental staff. DESIGN: Self-administered questionnaire. SUBJECTS AND METHODS: 204 NZDA members in Wellington, Otago, and Southland completed a self administered questionnaire about SCI. RESULTS: There has been little change in attitudes and practices of dentists towards SCI over the last decade. Few are actively involved in providing these services for their patients. Most dentists believe that there are significant barriers to providing SCI in their daily practice. CONCLUSION: Dentists require greater access to appropriate forms of training in SCI, and increased support (including resources) in order to increase the level of SCI provided in the dental setting. PMID- 15346878 TI - Antibiotic prophylaxis for at-risk patients. PMID- 15346879 TI - Modulation of dexamethasone-induced thymocyte apoptosis by heat-shock protein 90 binding agents. AB - Heat-shock protein 90 (HSP90) is known to affect a variety of cellular activities. The present study showed that the HSP90-binding agents, geldanamycin, herbimycin A and radicicol, inhibited the murine thymocyte apoptosis induced by dexamethasone and was accompanied by the inhibition of the reduction of the mitochondrial transmembrane potential (delta psi m). HSP90-binding agents did not inhibit etoposide-induced apoptosis. The inhibition of dexamethasone-induced apoptosis was in part due to the interference of HSP90 with the glucocorticoid receptor, resulting in the inhibition of nuclear translocation of the receptor. The expression of inositol 1,4,5-triphosphate receptors, which were shown to be involved in dexamethasone-induced apoptosis, did not participate in the inhibition of apoptosis. PMID- 15346880 TI - Neuromagnetic analysis of the late phase of the readiness field for precise hand movements using magnetoencephalography. AB - The aim of this study was to elucidate the cortical regulation of precise finger movements by using magnetoencephalography, with particular emphasis on the late phase of the readiness field. Magnetic brain signals were recorded non-invasively by 306 channel magnetoencephalography during the following two tasks. The first task, a simple task, was to bend the right thumb once as quickly as possible. The second task, a precise one, was to alternately oppose the thumb with the index finger and the middle finger of right hand. In this study, we confirmed that the differences between the two tasks were observed in the late phase of the readiness field, especially in the magnetic field 600 ms before the onset of movement. The activity of the magnetic field of the precise movement task was higher than the activity of the simple movement task. There were obvious differences in the spatial and temporal aspects of the left hemisphere. In the simple movement, the premotor area or motor area was activated in the late phase of the time window. The average latency from the EMG onset was -98.6+/-34.0 ms (n = 5). In the precise movement, the prefrontal area and the SMA were activated in the early and/or middle phases of the time window. The average latency from the EMG onset was -292.0+/-14.9 ms (n = 3) for the prefrontal cortex and -167.8+/ 38.3 ms (n = 4) for the SMA. The premotor area or motor area was activated in the late stage of the RF. The average latency from the EMG onset was -111.6+/-61.4 ms (n = 5). Many studies have been performed on the movement-related readiness field. However, the activity of the prefrontal area and the SMA had not previously been studied in the late phase of the readiness field. Our study indicated that the prefrontal area and the SMA played important roles immediately before the onset of precise finger movement. The integration of the prefrontal area, the SMA, and the premotor area is important for the onset of precise finger movement. PMID- 15346881 TI - Fluoride analysis of foods for infants and estimation of daily fluoride intake. AB - The mean daily fluoride intake in infants was estimated on the basis of their intake of commercial foods for infants in Japan and evaluated in order to establish the effectiveness and safety criteria for water fluoridation, which is practiced as a preventive measure for dental caries suitable in life stages from children to the elderly. Based upon the intakes of foods for infants, the mean daily fluoride intake was estimated to be 0.166 mg in infants aged 3-4 months, 0.202 mg in those aged 5-6 months, and 0.266 mg in those aged 7-8 months. The mean daily fluoride intake per kg of body weight at these ages was in the range of 0.023-0.029 mg/kg, which was about half of the standard daily fluoride intake for infants and children advocated by Ophaug et al., as 0.05-0.07 mg/kg. From our results, the daily fluoride intake of infants from foods in Japan is estimated to be equivalent to or lower than the values of previous reports in non-fluoridated areas. Consequently, our data support the argument that water fluoridation and the appropriate use of fluoride for dental caries prevention in Japan are needed on the basis of scientific criteria in terms of fluoride exposure related to food intake during tooth formation. PMID- 15346882 TI - Influence of template DNA degradation on the genotyping of SNPs and STR polymorphisms from forensic materials by PCR. AB - Detection of single nucleotide polymorphisms (SNPs) and short tandem repeat (STR) polymorphisms by PCR is widely used to analyze degraded DNAs in forensic science. The success of DNA analysis from human remains largely depends on the quality of the template DNA. We examined two SNPs (HLA-DQA1 and ABO) and two STR polymorphisms (VWA and CD4) by SSCP gel or denaturing gel electrophoresis, using two kinds of degraded DNA samples (165 teeth and blood stains contaminated with saliva) derived from the same person and investigated the influence of template DNA degradation on genotyping. As the degradation of DNA proceeds, unbalanced amplification of alleles occurred in the analysis of both SNPs and STRs, followed by allele drop, and further by loss of amplification. Non-target allelic products of STRs were amplified from highly degraded DNA samples; however, false allelic products of SNPs were not amplified from them. Amplification efficiency increased in proportion to the decrease of PCR target size, but reduction of the PCR target sizes also increased the chances of amplifying contaminating DNA, especially in highly degraded DNA specimens. The present results will help investigators to evaluate the genotyping of highly degraded DNA samples in forensic casework. PMID- 15346883 TI - Anti-phagocytic role of surface fibrous structure of an invasive Porphyromonas gingivalis strain. AB - Recent studies have shown that invasive and non-invasive strains of Porphyromonas gingivalis can both be isolated from patients with periodontitis. We examined the interaction between an invasive 16-1 P. gingivalis strain and phagocytes obtained from human peripheral blood and guinea pig peritoneal cavity. Phagocytes from human peripheral blood, mainly polymorphonuclear leukocytes (PMNs) isolated by centrifugation in Ficoll Hypaque, and macrophages collected from the peritoneal cavity of guinea pigs, were exposed to P. gingivalis cells. After this exposure, greater numbers of the non-invasive P. gingivalis ATCC 33277 were observed in human PMNs and guinea pig macrophages compared with the invasive P. gingivalis 16 1. Electron microscopic observations showed that invasive 16-1 within phagosomes in human PMNs and guinea pig macrophages retained their surface fibrous structures as well as their outer membranes. Electron microscopic examination showed that destruction and damage to the cell membranes and inner structures were clear in human PMNs and guinea pig macrophages after exposure to invasive 16 1 for 6 and 24 hours; this was a clear difference from exposure to the non invasive ATCC 33277. Release of lactate dehydrogenase (LDH) activities into the culture supernatant of PMNs after exposure to the invasive 16-1 for 4 and 6 hours was significantly greater than that after exposure to the non-invasive ATCC 33277 (p<0.05). On the other hand, the LDH activity after exposure for 21 hours to the invasive 16-1 was significantly lower than that of untreated cells and cells after exposure to the non-invasive ATCC 33277 strain (p<0.05). The PMN viabilities after exposure to cells of the invasive 16-1 for 3, 4, and 6 hours as evaluated by trypan blue staining were similar to those after exposure to cells of the non-invasive ATCC 33277, but that after exposure to the invasive 16-1 strain for 21 hours was significantly lower than that after exposure to cells of the non-invasive ATCC 33277 strain. PMID- 15346884 TI - Extracellular ATP both inhibits and facilitates calcium channel currents in acutely dissociated rat nucleus tractus solitarius. AB - The postsynaptic actions of exogenously applied adenosine 5'-triphosphate (ATP) were investigated in nucleus tractus solitarius (NTS) of the rat. Whole cell patch-clamp recordings were used to examine the regulation of voltage-dependent Ca2+ channels (VDCCs) currents (I(Ca)) by ATP in freshly dissociated NTS. Application of ATP inhibited I(Ca) from -905 pA to -741 pA. In addition to this inhibition, application of ATP facilitated I(Ca) from -941 pA to -1,094 pA in other neurons. The data presented here demonstrate for the first time that ATP has both inhibitory and facilitative effects on I(Ca) in NTS. It can be considered that ATP acts as a neurotransmitter in the NTS by having multiple regulatory effects on VDCCs. PMID- 15346885 TI - [Visualization of phosphoinositide: signaling lipids in living cells]. PMID- 15346886 TI - [Relationship between localized phosphatidylethanolamine exposure and yeast cell polarity]. PMID- 15346887 TI - [Visualization of glycosphingolipids on cells: visualization of lactosylceramide enriched glycosignaling domains of neutrophils]. PMID- 15346888 TI - [How to visualize cholesterol]. PMID- 15346889 TI - [Imaging lipid rafts]. PMID- 15346890 TI - [Single molecules view of lipid dynamics]. PMID- 15346891 TI - [Allelic exclusion of the odorant receptor genes]. PMID- 15346892 TI - [Transcriptional regulation of retinal photoreceptor cell development]. PMID- 15346893 TI - [Regulation of transcription in the Wnt signaling pathway]. PMID- 15346894 TI - [Artificial metallo-DNA: arraying metal ions using DNA]. PMID- 15346895 TI - [Prediction of the dispersion of birch pollen in Asahikawa]. AB - A birch pollen survey was carried out for 11 years from 1993 to 2003 in Asahikawa. An analysis was made of how the various meteorological factors related to the total birch pollen counts and the day when such pollen release started, and the correlation with and prediction of birch pollen dispersal were examined. The simple regression analysis provided evidence that the total birch pollen counts were related to the mean temperature in June of the preceding year. Also, the day on which birch pollen release commenced showed a correlation with the cumulative highest temperature of each day between March 21 and 31. These findings suggest that it is possible to predict both the total birch pollen counts and the first day of birch pollen release according to these meteorological factors. PMID- 15346896 TI - [Congenital laryngeal stridor]. AB - We reviewed the diagnosis, complications and treatment of congenital laryngeal stridor (CLS), in 97 patients who consulted our clinic between 1991 and 2001. The 97 patients were diagnosed with laryngeal malacia (32%), vocal cord paralysis and laryngeal stenosis (22%), a neoplastic disease like hemagioma and papilloma (11%), or cystic disease (7%). The cases with vocal cord paralysis, laryngeal stenosis or laryngeal cysts were usually diagnosed within 2 months of birth based on severe dyspnea. Two of the 31 cases of laryngeal malacia and 2 of the 22 cases of vocal cord paralysis were associated with neuromuscular disorders. Three patients suffered from vocal cord paralysis complicated by laryngeal stenosis. Thirty-three of the 97 cases required a tracheostomy; these 33 cases included the one case of laryngeal papilloma (100%), 9 of the 10 cases of hemangioma (90%), and 18 of the 24 cases of laryngeal stenosis (75%). Since any disorders of the upper airway can potentially induce stridor, establishing an accurate diagnosis is sometimes difficult when stridor is the only presenting symptom. Hence, information on associated symptoms and the past history of the subject is particularly important for an accurate diagnosis. In addition, decisions regarding the course of treatment course require adequate consideration of possible complications. PMID- 15346897 TI - [Clinical effect of bipolar radiofrequency thermotherapy on allergic rhinitis]. AB - The clinical effect of bipolar radiofrequency thermotherapy on allergic rhinitis was evaluated. A bipolar radiofrequency system (CelonLab ENT) was used to treat 16 patients suffering from allergic rhinitis between February 2003 and August 2003. The thermotherapy was performed under local anesthesia at the otolaryngology outpatient clinic of St. Marianna University Toyoko Hospital. Data were collected by questionnaire and rhinomanometry preoperatively and 2 months postoperatively. The mean visual analogue scale (VAS) score for intraoperative pain was 31 mm (range, 0-100), and nearly all the patients felt no or a subtle pain during the thermotherapy. Postoperative pain was also well tolerated, with nearly all the patients not requiring analgesic drugs. Postoperative bleeding was minor, and none of the patients required additional treatment for bleeding. Nearly all the patients reported an improvement in their nasal patency, rhinorrhea, headaches, and sleeping. Statistically significant improvements were observed for all the measured VAS scores: nasal patency, rhinorrhea, headache, and olfactory function. Nasal resistance, as measured by anterior rhinomanometry, significantly improved after treatment. The effect of decongestion was also measured using anterior rhinomanometry. The ratio of nasal resistance before and after decongestion was significantly higher after thermotherapy, suggesting that nasal decongestion had a smaller effect on nasal patency after treatment. The current results suggest that the CelonLab ENT device is an effective and safe treatment for allergic rhinitis. PMID- 15346898 TI - [Molecular biology of the ER stress response]. PMID- 15346899 TI - [Morphology and neurotoxicity of newly identified spherical beta-amyloid aggregates, 'amylospheroid', aiming at elucidation of the neurodegenerative cascades in Alzheimer's disease]. PMID- 15346900 TI - [Hepatocyte growth factor--history of discovery, physiological roles in dentistry and clinical application]. PMID- 15346901 TI - [Mass spectrometry for platelet biochemistry]. PMID- 15346902 TI - [Protein transduction and protection of cells against apoptosis]. PMID- 15346903 TI - [The role of molecular chaperones during protein transport into chloroplasts in higher plants]. PMID- 15346904 TI - [A SNP in BDNF gene is associated with human memory and BDNF secretion]. PMID- 15346905 TI - [Recent advances in Alzheimer's disease-related gamma-secretase complex]. PMID- 15346906 TI - [Noncompetitive immunoassays for small molecules]. PMID- 15346907 TI - Advances in Kawasaki disease. AB - Recent studies have increased our understanding of the etiopathogenesis of Kawasaki disease (KD). The inflammatory infiltrate in KD coronary artery aneurysms has been shown to consist of CD8 T lymphocytes, macrophages, and IgA plasma cells, consistent with an immune response to an intracellular pathogen with a mucosal portal of entry. The identification of an oligoclonal IgA response in the vascular wall and the detection of a KD-associated antigen in inflamed KD tissues using a synthetic antibody derived from KD oligoclonal IgA antibodies have provided new approaches to identification of the etiologic agent. Highly effective therapy has evolved for KD, even in the absence of identification of the etiologic agent. The existence of incomplete KD cases remains a significant diagnostic dilemma for the clinician. CONCLUSION: The development of a diagnostic test, more specific therapy, and ultimate prevention of this potentially fatal illness of childhood are dependent upon continued advances in determining the etiopathogenesis of this fascinating disorder. PMID- 15346908 TI - High versus restricted use of home oxygen therapy, health care utilisation and the cost of care in chronic lung disease infants. AB - Use of home oxygen therapy for prematurely born infants with chronic lung disease (CLD) can facilitate early discharge, but affected infants might require more readmissions. Our aim was to determine if health care utilisation and associated costs in the first 2 years were greater in centres with a high compared to centres with restricted use of home oxygen therapy. A retrospective review of the hospital and general practitioner (GP) medical records of 235 infants with CLD (median gestational age 27 weeks; range 22-33 weeks) was performed to note their readmissions, outpatient attendances, community service referrals and cost of care in the first 2 years after birth. A total of 76 infants (64%) in the high use centres and 12 (10%) in the restricted use centres were discharged home on oxygen. Infants in the high use centres were discharged home from neonatal care at a younger age (median 37.7 versus 39.9 weeks; P<0.001), but subsequently had similar numbers of inpatient events, and less GP (P =0.012) and community care (P < 0.001) contacts, although their duration of home oxygen use was longer (P < 0.001). The post-discharge costs were similar in the two types of centre, but the neonatal costs (P < 0.0001) and total cost of care per infant over the first 2 years (P < 0.0001) were lower in the high use centres. CONCLUSION: Early discharge and high use of home oxygen therapy was not associated with an increased cost of care or increased morbidity. PMID- 15346909 TI - Aquatic leech infestation: a rare cause of severe anaemia in an adolescent Tanzanian girl. AB - We report on a 15-year-old girl who presented with a history of chest pain, coughing, intermittent haemoptysis, fever, pallor and vomiting to a rural hospital in Tanzania. She was severely anaemic with signs of cardiorespiratory distress. Haemoglobin was 4.8 g/dl; the chest X-ray film was normal. She required one unit of blood. After 3 days, she vomited up a small parasite which proved to be an aquatic leech. Oesophagogastroscopy, then performed, revealed four small mucosal lesions in the pharynx and upper oesophagus. No other leeches were seen. Further oesophagogastroscopy 4 days later showed a normal mucosa. The girl was discharged on iron and folic acid supplements. A small lake near her village was identified as the likely source of her infestation. The family had used the water for drinking without any precautions-Terrestrial leeches can cause profound, life threatening anaemia, but even more so do aquatic leeches. This type is acquired while bathing or drinking unfiltered water. They attach themselves to mucous membranes, having been described in sites like conjunctiva, nose, pharynx/larynx, trachea/bronchi, oesophagus, vagina, and rectum. Especially in the airways, they can cause even fatal bleeding or obstruction. Treatment consists of removal of the leech. CONCLUSION: In developing countries, even unusual causes like leech infestation have to be considered in the differential diagnosis of severe anaemia in children. PMID- 15346910 TI - Oesophageal pH monitoring and reflux oesophagitis in irritable infants. AB - Acid reflux and/or oesophagitis may be responsible for inconsolable crying in infants. We evaluated prospectively the presence of acid reflux disease, oesophagitis and the accuracy of pH monitoring in the prediction of oesophagitis in a population of irritable infants. A 24-h oesophageal pH monitoring with a glass electrode and an upper gastro-intestinal tract endoscopy with grasp biopsies were performed in 60 irritable infants, aged 1 to 6 months, not responding to cow's milk elimination. The 24-h oesophageal pH monitoring was considered abnormal in 40/60 (66%) babies and histological oesophagitis was present in 26/60 (43%). In the infants with histological oesophagitis, the reflux index (% of the investigation time with a pH < 4.0) was > 5% in 18/26 (69%). Histology of the oesophagus was normal in 22 of the 40 (55%) infants with an abnormal pH monitoring. The mean reflux index in the group with oesophagitis (12.20%) was significantly higher than in the group with normal histology (8.74) (P = 0.036), although there was an important overlap. The sensitivity and specificity to predict oesophagitis with a reflux index of 5.0% or more was 69.2 and 35.3, respectively. There was not a reflux index which could be related to a clinically useful sensitivity and specificity to predict oesophagitis. CONCLUSION: Acid gastro-oesophageal reflux disease and/or histological oesophagitis were diagnosed in 66% and 43% of irritable infants, respectively. There was no relation between symptoms and abnormal pH metry or oesophagitis; however, the reflux index does not accurately predict oesophagitis and normal histology does not accurately exclude acid gastro-oesophageal reflux disease. Oesophageal pH monitoring and endoscopy provide additional information. PMID- 15346911 TI - Successful treatment with the monoclonal antibody rituximab in two children with refractory autoimmune thrombocytopenia. AB - Rituximab is a chimeric monoclonal antibody directed against normal and malignant mature B-lymphocytes and results in prolonged and severe B-cell depletion. Recently, rituximab has been successfully used in adult and paediatric disorders of B-lymphocytes such as autoimmune haemolytic anaemia and Werlhof disease. We report on two children with chronic immune thrombocytopenic purpura (ITP) refractory to steroids and immunoglobulins who achieved complete normalisation of their platelet counts after treatment with rituximab, 375 mg/m2 given weekly in four doses. In both cases the B-lymphocyte count dropped to zero after the second dose of rituximab and an unsupported platelet count > 100 x 10(9)/l was achieved during treatment. Six and 12 months after treatment, both patients remain well with normal platelet counts. CONCLUSION: This report supports the concept that rituximab may also be a valuable therapeutic option in children with chronic immune thrombocytopenic purpura refractory to standard treatment. Controlled clinical trials are needed to evaluate the efficacy and long-term side-effects of rituximab in this group of patients. PMID- 15346912 TI - Medical care of obese children and adolescents. APV: a standardised multicentre documentation derived to study initial presentation and cardiovascular risk factors in patients transferred to specialised treatment institutions. AB - So far in Europe, no studies have been published on the structuring of medical care for obese children and adolescents. Besides anthropometric parameters, evaluations of the cardiovascular risk factors hypertension, dyslipidaemia, impaired glucose metabolism and treatment modalities were documented in a standardised multicentre evaluation survey (APV) of 18 primarily outpatient and nine rehabilitation institutions. In total, 3837 children (aged 2-20 years) took part in the years 2000 up to March 2003, of whom 1985 were treated in outpatient institutions and 1852 in rehabilitation institutions. Of these children, 10% were overweight, 37% obese, 49% extremely obese and 4% of normal weight at initial presentation. The frequencies of diagnostic procedures performed and documented were low (measurement of blood pressure 43%, lipids 40%, glucose metabolism 21%). In the subgroup of obese children who were screened for cardiovascular risk factors, 23% suffered from hypertension, 11% displayed increased cholesterol, 9% increased low-density lipoprotein-cholesterol, 29% increased triglycerides, 11% decreased high-density lipoprotein-cholesterol and 6% had impaired glucose metabolism. CONCLUSION: Despite the high prevalence of cardiovascular risk factors in obese children and adolescents confirmed in this report, diagnostic procedures failed in a considerable percentage even in specialised treatment centres for obese children and adolescents. In future, the feedback based on standardised evaluation of diagnostic and treatment procedures should aim to improve the quality of medical care. PMID- 15346913 TI - Early deaths from childhood cancer. A report from the Childhood Cancer Registry of Piedmont, Italy, 1967-1998. AB - The population-based Childhood Cancer Registry of Piedmont (CCRP) has collected data on incidence since 1967. The occurrence of early death (i.e. within 30 days of diagnosis) was investigated in 3006 cases of childhood cancer diagnosed during the period 1967-1998. The proportion of early deaths (178 of the 3006 cases) was analysed by period of diagnosis (three decennial periods), age group, major diagnostic group and hospital category, with univariate statistics and logistic regression. The proportion of children with cancer who died within 1 month of diagnosis was 10.8%, 5.3% and 1.8% for cases diagnosed during 1967-1978, 1979 1988 and 1989-1998, respectively. This trend may reflect earlier diagnosis, improved diagnosis, more effective therapy or more frequent referral to specialised centres. The risk factors for early death were: age <1 year and diffuse disease at diagnosis, diagnosis during 1967-1978, a diagnosis of acute non lymphocytic leukaemia, non-Hodgkin lymphoma, central nervous system tumour or hepatic tumour. Early death was not related to the sex of the child. Care in an extra-regional specialised centre was associated with lower risk of early death. CONCLUSION: No temporal changes in early mortality were found among children with acute non lymphocytic leukaemia diagnosed in the first, second or third decade of activity of the CCRP, the percentages of children dying within 1 month being 12.8%, 10.7% and 12.8%, respectively. This pattern clearly differed from the corresponding trend for acute lymphoid leukaemia (6.4%, 2.0%, 0.4%). PMID- 15346914 TI - Severe isolated pulmonary Langerhans cell histiocytosis in a 6-year-old girl. AB - Langerhans cell histiocytosis (LCH) usually affects different organs or bones. Isolated pulmonary disease is rare in childhood. We report about a 6-year-old girl with progressive pulmonary insufficiency, onset of clubbing at 4 years of age and honeycombing lung infiltrations on X-ray films. The radiological suspicion of primary pulmonary LCH was confirmed by the presence of CD1a positive cells in the bronchoalveolar lavage fluid. Other organs were not involved. The girl was treated according to the LCH-III International Study Protocol with a good response. Follow-up showed no reactivation of LCH but a reduced vital capacity and signs of interstitial pulmonary involvement on a CT scan. CONCLUSION: Langerhans cell histiocytosis should be considered in the aetiology of cystic lung diseases. Early responders to treatment have a high likelihood of becoming free of disease. However, pulmonary fibrosis is an important mechanism of lung remodelling in pulmonary Langerhans cell histiocytosis and the long-term prognosis is unclear. PMID- 15346915 TI - Early diagnosis of Maroteaux-Lamy syndrome in two patients with accelerated growth and advanced bone maturation. AB - The diagnosis of mucopolysaccharidosis (MPS) VI is usually not made before the age of 2 or 3 years when the main clinical signs of dwarfed stature, skeletal deformities, coarsening facies, stiff joints and hepatosplenomegaly described in textbooks are recognisable. Conversely, accelerated growth with advanced bone age, a precocious feature of this condition although not suggestive of a storage disorder, is usually neither recognised, nor adequately interpreted. We report on two infants with MPS VI who presented with these two "unexpected" features within the 1st year of life. CONCLUSION: Recognition of precocious excessive growth in a mucopolysaccharidosis enables an early diagnosis, the prime responsibility of the clinician, in order to propose early treatment like bone marrow transplantation or active recombinant sulphatase therapy, and appropriate genetic counselling. PMID- 15346916 TI - Frequency of thrombocytopenia and large platelets correlates neither with conotruncal cardiac anomalies nor immunological features in the chromosome 22q11.2 deletion syndrome. PMID- 15346917 TI - Velocardiofacial syndrome presenting as distal arthrogryposis. PMID- 15346918 TI - Spinal muscle atrophy type 1 (Werdnig-Hoffman disease) with complex cardiac malformation. PMID- 15346919 TI - Association of oesophageal atresia, anophthalmia and renal duplex. PMID- 15346920 TI - Comparison of regional cerebral perfusion, EEG and cognitive functions in type 1 diabetic children with and without severe hypoglycaemia. PMID- 15346921 TI - Paediatric environmental health speciality units in Europe. For when? PMID- 15346922 TI - Evidence of a rapid decrease in prevalence of Helicobacter pylori infection in children of a high risk group living in Germany. PMID- 15346923 TI - Ulcerative colitis that developed 8 years after human parvovirus B19 encephalopathy. PMID- 15346924 TI - Aseptic meningitis in neonatal varicella complicated by Escherichia coil sepsis. PMID- 15346926 TI - [Tips on interactions with "difficult" patients. How "customer-friendly" is your practice?]. PMID- 15346925 TI - Congenital varicella syndrome. PMID- 15346927 TI - [About difficult patients and difficult physicians. Cookies do not help always]. PMID- 15346928 TI - [Acute abdominal pain in a Turkish patient after abdominal surgery. What has the surgeon overlooked?]. PMID- 15346929 TI - [Fennel tea and damp warm compresses. Prescriptions for the stomach and intestine]. PMID- 15346930 TI - [Source of pain or scapegoat in the epigastrium: how silent are gallstones?]. PMID- 15346931 TI - [Fear of a black man. When do children's fears become pathologic?]. PMID- 15346932 TI - [In skin changes often it is difficult to decide. Harmless growth or malignant tumor?]. PMID- 15346933 TI - [Scuba diving--jogging--mountaineering. We need more physical activity]. PMID- 15346934 TI - [The role of jogging in the prevention and treatment of cardiovascular disease]. AB - Lack of exercise and poor eating habits are considered to be major causes of most diseases of civilization. In consequence, endurance sports, but also an integration of physical activity in everyday life, are gaining in importance. Positive effects of regular physical exercise have been described for CAD, arterial hypertension, lipid metabolic disorders, type 2 diabetes mellitus and the metabolic syndrome. In order to achieve an optimal training effect, exercise intensity should be oriented to the individual anaerobic threshold. As a rule of thumb, 30 minutes of endurance training--ideallyevery day--is considered necessary. Prior testing of a person's ability to undertake such activities should include ECG and blood pressure measurements, spirometry and lactate determination, and contraindications must be taken into account. PMID- 15346935 TI - [Visiting high altitudes--healthy persons and patients with risk diseases]. AB - At altitudes higher than the threshold altitude of 2,500 m, high-altitude diseases may occur, usually after a delay of 6 to 12 hours. Apart from the headache associated with acute mountain sickness, life-threatening cerebral edema may develop. High-altitude pulmonary edema is a non-cardiac edema that often precedes acute mountain sickness. The most important preventive measure is a slow ascent. In the case of mountain sickness a prophylactic effect can be achieved with acetazolamide or dexamethasone possible, while for high-altitude pulmonary edema, nifedipine is the first-choice drug. Immediate descent and the administration of oxygen are always indicated. Patients with a high-altitude risk are those with cardiac or pulmonary disease. Nevertheless, it is still possible for patients with coronary heart disease, hypertension or bronchial asthma to attain to high altitudes. In contrast, patients with COPD, interstitial pulmonary disease or pulmonary hypertension are at appreciably greater risk. PMID- 15346936 TI - [Before going on a diving holiday--consider this]. AB - A prerequisite for all those who go in for the sport of scuba diving is mental and physical fitness. This must be checked within the framework of an examination for diving fitness before first taking up the sport, and should be rechecked every one to three years. For fitness for diving, not only lung function, ECG and tubal function need to be normal--the psychological profile of the holiday maker also plays an important role under water. For he or she must be able rapidly and reliably to recognize a dangerous situation and to react to it appropriately. Thorough training and the compliance with the rules of diving offer protection from many of the risks of this underwater sport--for example, problems with the ears or sinuses can be avoided. The most common cause of fatal diving accidents is a barotrauma of the lung with rupture of the alveoli on ascending. PMID- 15346937 TI - [Asthma control by utilizing the synergistic effect of inhaled steroids and long acting beta 2 agonists]. PMID- 15346938 TI - [Meteorologic influences in headache and migraine. There is something in the air]. PMID- 15346939 TI - [How does it function? Implantable cardioverter-defibrillator (ICD)]. PMID- 15346940 TI - [Isolation rate of gram negative microflora and its sensitivity to antibiotics in hemoblastosis patients]. AB - A total of 67 patients with blood system diseases and infectious complications were examined. During the period of the examination 139 microorganisms were isolated. Of these gram negative microorganisms constituted 51%, gram positive microorganisms--34.8% and fungal flora--14.2%. Most frequently the following gram negative microorganisms were isolated from the patients: Pseudomonas sp. (including P. aeruginosa), Klebsiella pneumoniae, Escherichia coli, Haemophilus influenzae. All isolated microorganisms retained sensitivity to imipenem, with the exception of individual strains of Pseudomonas sp.; the latter exhibited sensitivity to amicacin and ceftazidim. Cefotaxime was active with respect to 75% of K. pneumoniae strains and all E. coli strains, ciprofloxacin was active with respect to 43% of E. coli strains, 80% of K. pneumoniae strains and 83.4% of Pseudomonas sp. strains, cefepim was active with respect to 85.7% of Pseudomonas sp. strains and all E. coli strains, ceftazidim was active with respect to all Pseudomonas sp. and E. coli strains. 75% of K. pneumoniae strains, 77.8% of Pseudomonas sp. strains and 86% of E. coli strains retained sensitivity to amicacin. 25% of K. pneumoniae strains required testing for ESBL production. PMID- 15346941 TI - [Biological properties of microorganisms as the basis for the prognosis of the severity of pyoinflammatory pleuropulmonary diseases]. AB - In the study of 65 microbial cultures isolated from the purulent foci in acute pulmonary abscess and acute pyothorax of 48 patients, a wide spectrum of microflora was detected. Staphylococci and Pseudomonas prevailed among aerobes, bacteroids and peptostreptococci--among anaerobes. In cases of the prolonged course of the pathological process, as compared with the common one, microorganisms exhibited hemolytic activity and high antilysozyme and anticomplementary levels. These findings served as the basis for working out a mathematical model for the prognosis of the disease course with 95% probability. PMID- 15346942 TI - [Characterization of Yersinia pseudotuberculosis heat stable serovar specific polypeptides with the use of monoclonal antibodies]. AB - The capacity of Y. pseudotuberculosis to express serovar specific polypeptides with different specificity of antigenic determinants was proved with the use of monoclonal antibodies (McAb). For the first time Y. pseudotuberculosis O antigens were found to have heat stable protein components carrying linear epitopes complementary to serovar specific MaAb and ensuring the serological specificity of the infective agent. The possibility of improving intraspecific classification of Y. pseudotuberculosis and their differentiation from other pathogenic Yersinia on the basis of the capacity of these bacteria for synthesizing species and serovar specific proteins is substantiated. PMID- 15346943 TI - [The action of Yersinia pseudotuberculosis heat stable lethal toxin on the biosynthesis of protein in vitro]. AB - The results of the in vitro action of Y. pseudotuberculosis heat stable lethal toxin on the biosynthesis of protein are presented. The toxin was shown to inhibit the inclusion of exogenous amino acid into newly synthesized peptides. The degree of the inhibition of the biosynthesis of protein in the in vitro system depends on the amount of the toxin added to the incubation mixture. The use of the conjugated transcription and translation system confirms our earlier data on the influence of the lethal toxin on the biosynthesis of protein in eukaryotic cells in vivo. PMID- 15346944 TI - [Influence of volatile metabolites of saprophytic soil microflora on the propagation of pathogenic bacteria]. AB - The influence of volatile metabolites of saprophytic soil microflora on the propagation of Listeria monocytogenes and Yersinia pseudotuberculosis is shown. Different character of interspecific relationships between bacteria, influencing their propagation, can be observed on the metabolic level. Volatile compounds produced by microorganisms are capable to act as both intra- and interspecific regulators of microbial communities. In this connection the propagation of pathogenic bacteria inhabiting soil may be stimulated or inhibited by the metabolic products of soil microorganisms. Methanol released by saprophytic bacteria into the environment play an important role in this process. PMID- 15346945 TI - [On the epidemiological situation in quarantine, natural focal and other infections on the territory of the Southern Federal District]. AB - The data on the sanitary and epidemiological situation in the Southern Federal District are presented. The analysis of morbidity in tuberculosis, measles, HIV infection, viral hepatitis A, typhoid fever, cholera and quarantine infections, Crimean hemorrhagic fever, West Nile fever, rabies, malaria has been carried out. Special attention has been given to "new and newly returning infections", and among them to the spread of SARS ("atypical pneumonia"). The role of regional epidemiological safety programs, in particular such program as "The prophylaxis of quarantine and natural focal infections and the sanitary protection of the territory of the Southern Federal District of the Russian Federation from the import and spread infectious diseases in 2003-2005", has been substantiated. PMID- 15346946 TI - [Molecular epidemiological analysis of the spread of human immunodeficiency virus (HIV) infection in the Novosibirsk region]. AB - The dynamics of the spread of individual subtypes of type 1 HIV (HIV-1), circulating in the Novosibirsk region during the epidemic rise of HIV infection was under study. The epidemic of HIV-1 in Novosibirsk has a pattern similar to that in Russia as a whole. At the initial stage of epidemics multiple sources of virus determine the heterogeneity of the isolated subtypes of HIV-1. Then the parenteral route of HIV transmission, connected with the intravenous use of narcotic drugs, becomes dominant. Recently the spread of HIV-1 from the group of intravenous drug users to other groups of the population has been observed. In the circulation of HIV-1 among drug users the leading role was shown to belong to subtype A, which ensures its rapid spread and dominating role in the epidemic process. Further spread of the HIV-1 epidemic is expected to proceed in parallel to the spread of viral hepatitis, sexually transmitted diseases and drug addiction. Thus, HIV-1, subtype A, may be assumed to be dominant in the Novosibirsk region in the nearest future. PMID- 15346947 TI - [Immunostimulating activity of polycomponent vaccine "Immunovac-VP-4" and grippol after their combined administration]. AB - The experimental study of the immunostimulating activity of therapeutic bacterial polycomponent vaccine VP-4 and prophylactic vaccine grippol, introduced both separately and in combination, on mice infected with Salmonella typhimurium, used as a model. Both preparations were found to produce an immunomodulating effect. The combined subcutaneous injection of VP-4 and grippol did not decrease their immunostimulating activity, but their separate administration at an interval of 14 days resulted in essential decrease in the protective activity of each of these two preparations. As shown on the model of Klebsiella infection in mice, challenged 4 weeks after immunization, VP-4 ensured the survival of 78.6% of mice, while after the injection of grippol their survival rate was not different from that of the group of intact animals. The evaluation of the immunostimulating activity of these preparations under the conditions of the prophylaxis of influenza and acute respiratory infections in organized groups of children revealed that the use of VP-4 alone or grippol in combination with VP-4 considerably decreased the number of secondary bacterial complications in children. PMID- 15346948 TI - [Use of therapeutic polycomponent vaccine "Immunovac VP-4" for the treatment of chronic bacterial prostatitis]. AB - A complex of clinical and laboratory examinations of patients with bacterial chronic prostatitis was carried out. The patients were divided into two groups: 55 patients treated by the method of immunotherapy and 20 patients (controls) undergoing basic therapy. Immunotherapy was carried out with the use of polycomponent vaccine VP-4 containing Staphylococcus aureus, Klebsiella pneumoniae, Proteus vulgaris and Escherichia coli antigens. Vaccinal therapy was indicated in cases with infection focus associated with opportunistic bacteria, a prolonged torpid course of prostatitis and when different schemes of etiotropic therapy proved to be ineffective. The vaccine was introduced in 4 or 5 subcutaneous injections at an interval of 5-7 days. The study revealed that in cases of chronic prostatitis accompanied by microecological and immune disturbances the use of complex therapy allowed to achieve good clinical and laboratory results in 91% of the patients. In the patients undergoing immunotherapy the elimination of opportunistic bacteria from the urogenital tract and the restoration of affected microflora occurred. PMID- 15346949 TI - [Immune response to the oral administration of a new meningococcal preparation, serogroup A, under experiment]. AB - In the experiment on rabbits immune response to the oral administration of a new Neisseria meningitidis whole culture preparation, serogroup A, was demonstrated. The preparation was based on the acetone fixed culture, grown by the continuous flow method under a computer-controlled constant level of oxygen. The immunological activity of the preparation was demonstrated. In the blood sera of rabbits examined by immunoenzyme assay and the passive hemagglutination test, a multiple increase in the content of hemagglutinating and IgG antibodies to polysaccharide, outer membrane proteins and lipooligosaccharide was noted, their content remaining at a high level for 303 days (the term of observation). The oral immunization with the preparation protected mice infected with N. meningitidis live culture, serogroup A. PMID- 15346950 TI - [Clinical and immunological aspects of cervicitis of chlamydial etiology]. AB - A total of 52 women with established presence of the causative agent of chlamydial infection in the cervical canal were examined. The chlamydial lesion of the cervix uteri was characterized by the absence of clinical manifestations in 25% of cases, faintly pronounced local inflammatory process in the lower part of the genitals in the form of chronic cervititis in 57.7% of cases and the symptoms of subacute cervicitis in 17.3% of cases. In case of a chronic course of chlamydial cervicitis a low content of cytokines IL-1alpha, IL-1beta, TNF-alpha and an elevated concentration of IL-8 were observed. In the cervical slime of patients with chlamydial cervicitis no changes in the total content of leukocytes were noted in comparison with healthy women, but the number of nitro blue tetrazoliu reducing neutrophils increased, while their functional reserve and phagocytic activity dropped. Autoneutrophilokins used for local immunocorrection in the treatment of women with chlamydial cervicitis produced a normalizing effect on the local infectious protective factors of cervical secretion, thus enhancing the effectiveness of therapy. PMID- 15346951 TI - [Dynamics of humoral immune response to Treponema pallidum proteins p17 and p41 at early stages of syphilis]. AB - In 60 blood sera from syphilis patients the titers of IgG to T. pallidum antigens p17 and p41 were detected with the use of the test system based on the recombinant analogues of T. pallidum proteins. The study revealed that primary syphilis was characterized by considerable prevalence of IgG to protein p41 with the total antibody level being low, while early latent syphilis was characterized mainly by considerable prevalence of IgG to protein p17 in the presence of high titers of antibodies. In secondary syphilis the sera contained a high total antibody level and a wide range of IgG ratios to individual antigens. On the basis of the data obtained the dynamics of immune response to antigens p17 and p41 at the early stages of the disease was hypothetically plotted. The curves of antibody levels had a wave-like character with the phase shifts of peaks for individual proteins and very low antibody titers (less than 1:100) in the negative peak areas. Conclusions were made that it was necessary to use the mixture of antigens in the production of the test systems and, when designing reference panels of sera, to include sera with extremely low titers of antibodies to individual proteins. PMID- 15346952 TI - [Laboratory diagnostics of enterovirus cardiac infections]. AB - Diagnostic efficacy of different markers of enterovirus cardiac infection (EVCI) has been evaluated. Testing of clinical samples from patients with myocarditis (n=50), dilatation cardiomyopathy (n=122), ischemic heart disease (n=34) and from healthy donors (n=50) revealed diagnostically significant markers in patients suspected for enterovirus cardiac infection: antienterovirus IgM in the patient's blood serum, the expression of viral proteins by myocardium cells and the presence of genome RNA and replicative intermediate in cardiac cells. The results obtained were used for developing up-to-date scheme of the EVCI diagnosis which included the data from the case history, the preliminary rapid diagnosis and the molecular biological study of the cardiac biopsies. PMID- 15346953 TI - [Detection of bacterial molecular markers in the tissue of cardiac valves in normal and pathological states by gas chromatography and mass spectrometry]. AB - Samples from cardiac valves of 31 patients were analyzed by gas chromatography and mass spectrometry. The algorithm of mass spectrometric parameters was developed, which permitted the determination of about 200 known microbial fatty acids, aldehydes and sterols, sufficient for the detection and quantitative determination of more that 170 taxons of clinically significant microorganisms on the genus or species levels. The quantitative and qualitative differences in the composition of microbial markers of endocardial valves in normal and pathological states, particularly in cases of infectious endocarditis, were detected. The participation of 37 microbial taxons in the process was confirmed. The level of endocardium colonization in infectious endocarditis reached from 2 to 7 x 10(9) microbial cells/g of valvular tissue (which exceeded twofold the equivalent concentrations of the marker in the normal state). In terms of quantity, the leading role was played by Cardiobacterium hominis. PMID- 15346954 TI - [Immunoserodiagnosis of acute Streptococcus pneumoniae and Chlamydia pneumoniae infections in the period of epidemic rise of non-hospital pneumonia in children in St. Petersburg, 1998-2001]. AB - The dynamics of the antibody formation to S. pneumoniae and C. pneumoniae in children during the epidemic outbreak of non-hospital pneumonia in St. Petersburg in 1998-2001 was studied. For the first time the inhibiting influence of acute C. pneumoniae infection on the synthesis of antibodies to S. pneumoniae in acute mixed infection was established. The prolonged (up to days 29-39 of the disease) circulation of IgM and IgG antibodies in acute chlamydial infection, as well as the prevalence of the primary infectious process, were detected. PMID- 15346955 TI - [Clinical and diagnostic importance of the evaluation of the Ig proteases activity in children with intestinal dysbacteriosis]. AB - The specific activity of serine, metal dependent and thiolic Ig proteases in the coprofiltrates of children with manifestations of intestinal dysbacteriosis was determined by the enzyme immunoassay. 56 children with pronounced symptoms of intestinal disorders (37 children aged up to 1 year and 19 children over 1 year) were examined. A group of 25 clinically healthy children was used as control. Simultaneously with protease activity of coprofiltrates, there was detected the level of Ig-degrading activity of the opportunistic bacteria islolates of different taxonomic groups from feces of children with dysbacteriosis of different severity (as determined by the classical bacteriological method). The evaluation of the Ig-proteolytic activity of fecal supernatants, associated with the presence of serine, metal-dependent and thiolic proteases in the intestine, as well as detection of such proteases in microbial isolates, seems to be highly important for the diagnosis of intestinal disorders in children and is recommended for screening of intestinal dysbacteriosis. PMID- 15346956 TI - [Role of iron ions in the production of hemolysin by toxigenic and nontoxigenic Vibrio cholerae of different serogroups]. AB - The hemolytic activity of ctx- and ctx+ V. cholerae, serogroups eltor and O39, in a medium free of FeCl3 was studied. During the cultivation in this medium, the strains of both V. cholerae serogroups proved to be capable of lysing sheep red blood cells in the Graig test, irrespective of the presence of ctx genes. The cultivation of V. cholerae ctx+ strains of both serogroups under such conditions facilitated the production of hemolysin with the same spectrum of lytic activity as hemolysin produced by ctx- strains. PMID- 15346957 TI - [Heterogeneity of Bacillus anthracis strains in terms of their adhesive capacity]. AB - The homogeneity of colonies of two B. anthracis vaccine strains in R- and RS- forms (100 colonies of each strain) in terms of their adhesive capacity was studied. B. anthracis strain 228/8 showed more heterogeneous composition than B. anthracis strain 71/12, moderately and highly adhesive colonies prevailing in both forms and nonadhesive colonies being absent. The prevalence of highly adhesive clones was established in the RS- form of B. anthracis strain 72/12 in comparison with R- form. By the average value of the adhesion index the RS- form colonies of this strain were classified as highly adhesive, while the colonies in the R- form were characterized as moderately adhesive. PMID- 15346958 TI - [Antibody affinity in serological immunosuspension tests]. AB - An original method for the determination of antibody affinity (in SI physical units) with the use of serological immunosuspension tests--the indirect hemagglutination (HA) test and the latex agglutintion (LA) test--was developed. The immunological and physico-chemical properties of suspensions in the indirect HA test and the LA test were linked with the character of the manifestation of test results in the form of "umbrellas" or "buttons" which appeared as the result of the sedimentation of physical carriers. The experimentally determined mass of carriers per unit of volume of the test suspension made it possible to establish bioenergy per unit of volume by the potential energy of carriers forming "umbrellas". Antibody affinity is the force of interaction in one pair of determinants, expressed in newtons and determined by means of the indirect HA test and the LA test. Thus, in antibodies to the causative agent of plague it was, respectively, 1.028x10(-17) and 0.1014x10(-17). PMID- 15346959 TI - [Phenotypical analysis of heterogenic Vibrio cholerae strain Dacca 35 Ogawa]. AB - The comparative analysis of the production of the main pathogenicity factors by toxigenic and non-toxigenic clones of V. cholerae natural classical strain Dacca 35 Ogawa has been carried out. The data obtained in this analysis indicate that the appearance of turbid colonies, not synthesing cholera toxin, is linked with the production of an exopolysaccharide layer on the outer surface of the cells, which determines their morphology. The suggestion has been made that the regulatory gene toxR controls the expression of not only cholera toxin, protein OmpU, but also exopolysaccharide. PMID- 15346960 TI - [Spread of bacteria of the genus Acinetobacter in the hydrobios of the Bay of Peter the Great, the Sea of Japan]. AB - The analysis of the spread of bacteria of the genus Acinetobacter among the invertebrate animals of the Bay of Peter the Great (the Sea of Japan): in bivalved mollusks, sea cucumbers and sea urchins Acinetobacter bacteria were detected more often than in cushion stars and crustaceous animals. The biological properties of 45 isolated bacteria were studied with the use of a wide variety of tests. The study revealed that 9% of Acinetobacter cultures isolated from marine animals had pronounced beta-hemolysis. PMID- 15346961 TI - [Neutrophils and antioxidant activity of cervical secretion in women with the acute inflammation in the upper section of the reproductive tract]. AB - The functional activity of neutrophils, peroxidation products and antioxidative activity of cervical secretion in women with the acute inflammatory process in the upper section of the reproductive tract were under study. The most pronounced changes in the function of neutrophils were observed in women with the low antioxidative activity of cervical slime. PMID- 15346962 TI - [Relationships between the content of circulating immune complexes, the concentration of serum immunoglobulins and the titers of specific antibodies in patients with tick-borne encephalitis]. AB - In the blood serum of tick-borne encephalitis (TBE) patients the detection rate and concentration of circulating immune complexes, as well as the content of serum IgA, IgM and IgG, were evaluated. The formation of immune complexes was found to depend on IgM and IgG specific antibodies to TBE virus, the period of the disease and the clinical form of virus infection. PMID- 15346963 TI - [Decreased resistance of multiresistant mycobacteria to isoniazid during the treatment of experimental tuberculosis with ozone and isoniazid]. AB - Mycobacteria (MB) of the clinical strain resistant to streptomycin, isoniazid (IN), rifampicin and kanamycin were injected intravenously into 68 BALB/c mice. The animals were divided into 5 groups: two control groups 0 and 1 (intact and infected without subsequent treatment), group 2 (treated with IN), group 3 (treated with IN and injected intraperitoneally with dissolved ozone, or dO3), group 4 (injected with dO3). The animals started to die by month 4 after the infection. By month 5 all mice died with the exception of intact mice and those treated with dO3). By month 4 the study of MB cultures isolated from the lungs revealed a decrease in their resistance to IN in the groups undergoing treatment with dO3. Hepatic and splenic lesions were observed after treatment with IN only were greater than in the absence of treatment. After the use of IN + dO3 such lesions were the least. The mechanism of a decrease in the medicinal resistance of MB under the action of dO3 and the expediency of the simultaneous use IN and dO3 in cases of the unknown medicinal resistance of MB are discussed. PMID- 15346964 TI - [Molecular aspects of the damaging action of bacterial lipopolysaccharides]. AB - The review deals with the molecular aspects of the interaction of bacterial lipopolysaccharides (LPS, with the macroorganism: the sources of LPS in the organism, its detoxication, regulation disturbances in pathology (septic shock). Special attention is drawn to the complex functioning of cytokines in the development of the inflammatory process both at the initial stage, characterized by the initiation of the cytokine cascade and at the final stage resulting in convalescence or the development of pathology. The most promising trends of research in this field are discussed. PMID- 15346965 TI - [Role of proteins of the macroglobulin family in the mechanisms of infection]. AB - Information on the properties of proteins of the macroglobulin family taking part in the host protection from viral, fungal and bacterial pathogens is reviewed. High plasticity and polyfunctional character of these proteins makes it possible to realize different protective functions. They inhibit the lysis of the cell wall by binding the hydrolases of the pathogen thus blocking its penetration into the cell, directly participate in the presentation of antigens to immunocompetent cells, transport antibacterial substances (interferons, lysozyme) to the zone of infection. In addition, macroglobulins take part in the apoptosis regulation in infected cells, utilization of the lysosomal enzymes of annihilated pathogens. The complexes of macroglobulins with some proteins are powerful inductors of antibody production. Further studies of the properties of these proteins will result in a better understanding of the nature of infectious process. The possibility of artificial formation of macroglobulin complexes with pathogen components or with substances possessing protective or anti-inflammatory properties opens prospects for using these proteins in the fields of vaccinology, gene therapy and molecular biology. PMID- 15346966 TI - [Seasonal distribution of the epidemic outbreaks of enteric infections as an indicator of the seasonal activity of the epidemic process]. PMID- 15346967 TI - [Isolation of Chryseomonas luteola in a patient with combined trauma]. PMID- 15346968 TI - [Natural foci of leptospirosis and hemorrhagic fever with renal syndrome on the territory of the Ulyanovsk region]. PMID- 15346969 TI - Beyond surveys and focus groups: usability testing can help you hone your Web site design. AB - Usability testing goes beyond surveys, Web trend analysis, and focus groups by providing direct observation of how people interact with a site. The process can be informal and inexpensive, involving a few participants and a Web designer. Or it can be more formal, using special technology and professional usability analysts. But even a relatively casual usability test can give Web designers information about the site that they can't get in any other way. PMID- 15346970 TI - MemorialCare provides expanded physician directory entries. PMID- 15346971 TI - Drug company's consumer health portal encourages return visits. AB - How do you get consumers to return to your site? For some health-site surfers, the offer of a portal where they can customize pages and store their own data may have so much appeal that they'll make it their home on the Web. But to really win their loyalty, a consumer portal has to offer a lot of features that will help them manage their health activities. A good example is My Health Zone, launched by pharmaceutical company Schering-Plough. PMID- 15346972 TI - Report details eRx challenges and provides suggestions for increased adoption. AB - A new report by the eHealth Initiative (http://www.ehealthinitiative.org) looks at electronic prescribing, or "eRx," and discusses its benefits, challenges to adoption, and movements toward adoption. Released in April 2004, the 154-page report--Electronic Prescribing: Toward Maximum Value and Rapid Adoption--also highlights recommendations for the optimal design of an eRx system. PMID- 15346973 TI - [Acute recurrent pancreatitis: how to direct oneself in the maze of multiple etiologies and complementary tests?]. AB - Acute recurrent pancreatitis are troublesome events in life's patients, because of possible complications, and fear of recurrency and pain. Then, finding the appropraited etiology is the only way prevent recurrency. PMID- 15346974 TI - [Acute and recurrent night leg pain in young children: "growing pains"]. AB - Acute nocturnal and recurrent pain of the limb in children or "growing pains" constitute a poorly known pathological entity. Growing pains occur in 20% of children aged 2 to 12. The diagnosis is made after ruling out all organic pathology: inflammatory, infectious, post-infectious, traumatic, metabolic, tumoral, vascular, hematologic, orthopaedic and benign pathology of children. The treatment requires analgesics and soft massage. PMID- 15346975 TI - [Genetics of migraines: from ionic channels to single nucleotide polymorphisms?]. AB - Our knowledge about migraine pathogenesis has increased exponentially over the last decade and this greatly due to the advances in genetics. In familial hemiplegic migraine (FHM), the findings of mutations in the CACNA1A gene (19p13), coding for the pore-forming subunit (alpha1A) of neuronal voltage-dependent P/Q type calcium channels (FHM1), and in the ATP1A2 gene (1q21-23), encoding the alpha2-subunit of the Na+, K+ ATPase ionic pump (FHM2) have focused attention on central nervous system ionic channels and helped to better understand FHM pathophysiology. A dysfunction of these channels modifies neuronal excitability (favouring spreading depression), chemical neurotransmission and, indirectly, neuronal metabolism. These channels may represent targets for novel anti-migraine drugs, which underscores their importance for the frequent forms of migraine (without or with aura). Studies of gene associations, neuromuscular transmission, cerebellar functions, neuronal excitability and metabolism and certain drug effects suggest indeed that ionic channels play a pathogenic role in migraine with aura patients. However, in the majority of patients they are probably not the sole culprit, since most of the frequent forms of migraine seem to have a more complex genetic predisposition based on a number of single nucleotide polymorphisms. The challenge for the next decade is to establish correlations between the geno- and the phenotype of migraine patients which needs more frequent and focused genetic studies and a more precise phenotype, based on clinical as well as on neurophysiologic and metabolic data. PMID- 15346976 TI - [Raynaud's phenomenon]. AB - Raynaud's phenomenon (RP) is a common vascular disorder characterized by a recurrent transient vasospasm of the fingers and toes on exposure to cold or with emotional stress. Clinical criteria are used to distinguish patients with primary or idiopathic RP (formely Raynaud's disease) from those with secondary RP (formely Raynaud's syndrome). They include history, general physical examination, capillaroscopy, test for antinuclear antibody and erythrocyte sedimentation rate. More specific exams may be needed in selective cases. Excluding RP of occupational origin, the most common cause of secondary RP is a connective tissue disease (particularly scleroderma). RP may precede other clinical symptoms by several years, but most patients with RP will not progress to systemic disease. Nevertheless, primary RP may cause significant disability. We review the pathophysiology and clinical criteria of the disorder. Unfortunately, management of PR is still empirical and largely supportive. PMID- 15346977 TI - [New advances in drug therapy in 2003-2004]. AB - The most important drugs registered and/or launched in Belgium during the last year in the various disciplines of internal medicine will be briefly described. The main characteristics of each molecule and its modalities of appropriate use in clinical practice will be emphasized. Both the efficacy and safety of these new drugs have been evaluated in randomised controlled trials according to the evidence-based-medicine criteria. Most of them are currently studied in order to further demonstrate their potential benefit on a long-term basis, in large cohort of patients and using hard clinical endpoints. PMID- 15346978 TI - [How I treat... the critical bacterial colonization of a leg ulcer. The Yin and the Yang features of silver-based dressings]. AB - The critical bacterial colonizaion of leg ulcers can impair their healing rate, aggravate the patient discomfort and increase the medical and nursing costs. In recent times, the dressings designed for leg ulcers have followed a pace of conceptual revolution. Some of them are now offered containing an antiseptic of the silver salt family. The silver concentraton delivered into the wound bed is important to consider when assessing treatment efficacy. The diversity of the silver-based dressings currently on the European market is as large as their differences in activity. Only a minority of these dressings adequately control the wound biocenosis. Their cost which is high for the patient, must be compared to that of nursing care that may become less important. The expected beneficit is a reduction in healing time. PMID- 15346979 TI - [Optimizing the managment of patients with diabetes mellitus: selected clinical trials from the 2004 Congress of the American Diabetes Association]. AB - The 64th scientific congress of the American Diabetes Association had a special session devoted to the presentation of the results from three clinical trials: 1) the first multicentre international trial of pancreatic islet transplantation according to the so-called Edmonton protocol with the primary endpoint of restoring insulin independence in type 1 diabetic patients; 2) three pivotal studies of 30 weeks testing both the efficacy and safety of exenatide (exendin 4), a new insulin secretagogue that is a long-acting analogue of glucagon-like peptide-1, in patients with type 2 diabetes treated with either metformin, or a sulfonylurea, or a metformin-sulfonylurea combination; and 3) the "Collaborative AtoRvastatin Diabetes Study" (CARDS), a placebo-controlled primary prevention trial of cardiovascular complications using atorvastatin 10 mg in 2 838 at risk patients with type 2 diabetes. The main results and conclusions of these trials are briefly presented as they open new perspectives in the management of patients with type 1 or type 2 diabetes mellitus. PMID- 15346980 TI - [Study on the role of calcium signal during mature resumption of isolated mouse germinal vesicles in a cell free system]. AB - A cell-free system including HeLa cell lysate of synchronized metaphase or G2 phase and isolated germinal vesicles (GV) from mouse oocytes was used to study the role of calcium and its downstream mediator during mature resumption. The isolated GVs could resume meiotic maturation in the lysate prepared from M phase HeLa cell, which marked by chromatin condensation. And this process was not affected by calcium chelating agent. But calcium in lysate from G2 phase cells was critical to meiotic maturation. Only in mid-G2 phase cell lysate (released from nocodazole for about 20-23h) chromatin condensation could be induced by calcium. Calcium had no effect on the cell lysate prepared from earlier (18-20h) and later (24h) G2 phase cells. Further studies showed that down stream mediator CaM and CaMKII might also involove in this process. Inhibition the function of CaM and CaMKII could block GVBD and first polar body extrusion of DOs cultured in vitro. The target of calcium signal might be MPF because MPF was existed from mid G2 phase to metaphase and the tyrosine phosphorylation level of Cdc2 subunit was significantly dephosphorylated in M phase. Our results further confirmed that the resumption of meiosis maturation was promoted in a calcium/CaM depended pathway. PMID- 15346981 TI - [Isolation of ABA-regulated genes in Oryza sativa through fluorescent differential display PCR (FDD-PCR)]. AB - Abscisic acid (ABA) was critical in plant seed development and response to environmental factors such as stress situations. To study the possible ABA related signaling transduction pathways, we tried to isolate the ABA-regulated genes through fluorescent differential display PCR (FDD-PCR) technology using rice seedling as materials (treated with ABA for 2, 4, 8 and 12h). In the 17 fragments isolated, 14 and 3 clones were up-and down-regulated respectively. Sequence analyses revealed that the encoded proteins were involved in photosynthesis (7 fragments), signal transduction (1 fragments), transcription (2 fragments), metabolism and resistance (6 fragments), and unknown protein (1 fragments). 3 clones, encoding putative alpha/beta hydrolase fold, putative vacuolar H+ -ATPase B subunit, putative tyrosine phosphatase, were confirmed to be regulated under ABA treatment by RT-PCR and northern blot analysis. FDD-PCR and possible functional mechanisms of ABA were discussed. PMID- 15346982 TI - [Genetic analysis on diversity of reproductive modes of silver crucian carp inferred by transferrin and isozyme markers]. AB - Silver crucian carp (Carassius auratus gibelio) has been known for its gynogenetic reproduction. In this paper, eggs from clone F of gynogenetic silver crucian carp were inseminated respectively by sperms of clone D, clone A and common carp, and polymorphic patterns of transferrin and isozymes of the produced three offspring FD, FA and FL were studied comparatively to explore the diversity in reproductive modes of silver crucian carp. As control, zymograms of FL progenies exhibited a maternally clonal inheritance, and gynogenesis was reconfirmed. However, differentiation of morphs and electrophoretic patterns was both observed among the FD progenies, and detection of bands specific for the clone F or clone D in some FD individuals urged the occurrence of recombination. Furthermore, extreme linkage disequilibrium for different protein loci suggested that linkage groups composed of different genes might function as the fundamental unit in the recombination. With respect to FA group, phenotypes of parental clones were both detected in the F2 generation (FA x FA progenies) while only maternal phenotypes were detected in the F1 generation. It appeared that both the parental chromosome sets could be transmitted to the offspring but expression of paternal genes were fundamentally upset in the F1 generation because of incompatibility of the parental genes. Generally, genetic analysis of FD and FA offspring primarily exhibited a particular syngamy for silver crucian carp. Besides the clonal reproduction of gynogenesis, syngamy could provide opportunities of recombination for silver crucian carp, which might remove some genetic loads from the genome and introduce new genotypes. Diversity in reproductive modes might play an important role in ecological adaptation of silver crucian carp. PMID- 15346983 TI - [Relationship between terminal sialyl and fucosyl residues of glycans on cell surface and cell biological behaviors]. AB - The relationship between the structures of glycans on the surface of H7721 cells, a human hepatocarcinoma cell line, and the cell behaviors was studied by using neuraminidase and alpha-L-fucosidase to remove the terminal sialyl or fucosyl residues of surface glycans respectively. The cell adhesion to fibronectin (Fn), laminin (Ln) and human umbilical vein epithelial cell (HUVEC), as well as cell chemotactic migration and invasion were selected as the parameters of the cell behaviors. It was found that sialyl residue was not essential for the cell adhesion to Fn, but was important in the cell adhesion to Ln and chemotactic cell invasion, and very crucial in the cell adhesion to HUVEC and chemotactic migration. In contrast, fucosyl residue was probably participate in cell adhesion to Fn, Ln and HUVEC, but not important in chemotactic migration and invasion. The cell adhesion to HUVEC, chemotactic migration and invasion were inhibited by the monoclonal antibody of sialyl Lewis X (SLex), but not by the antibody of non sialyl Lewis X (Lex). This result supports the finding that sialyl residue is more important than fucosyl residue in the contribution to the above-mentioned three cell processes. PMID- 15346984 TI - [Study on the cloning goat by cross-breed somatic cell nuclear transfer]. AB - Shaneng goat is a famous milking species. Boer goat is world famous goat breed for creophagism. In this study, we evaluated the development potential of adult Boer goat's somatic nuclei after nuclear tansfer (NT) into enucleated MII oocytes of the Shaneng goat. Somatic donor cells were obtained from two different sources: 1) adult granulosa cells (GCs) and 2) adult skin fibroblasts (FCs). The reconstructed embryos that developed to morula or blastocyst stage in vivo were transferred to 38 synchronized recipient. CONTROL: Somatic donor fibroblast cells were obtained from a fetal at 35 day. In the same way the reconstructed embryos were directly transferred to synchronized recipient of Shaneng goats. (1) Experimental group: NT embryos derived from GC and FC developed into morulas and blastocysts at a frequency of 46.8% and 31.4% respectively. Fifty-two NT morula and blastocyst stage embryos were transferred in to 38 recipients, Three of which were confirmed to be pregnant (7.9%). All pregnancies were not maintained to term. (2) CONTROL group: 136 NT embryos were transferred in to 14 recipients, Six of which were confirmed to be pregnant (42.9%). Four of those were maintained to term. Four recipients delivered four male kids (2.9% of embryos transferred). One male kid died at birth, the dead lamb shows as "large offspring syndrome". the others appeared health and normal. DNA analysis confirmed that those kids were genetically identical to their donor. These results demonstrated that Shaneng goat somatic cells could direct normal development and Shaneng goat oocyte cytoplasm supported development of preimplantation embryos produced by NT of somatic cell nuclei from Boer goat. PMID- 15346985 TI - [The ultrastructures of storage protein accumulation in aleurone layer and cotyledons of Fagopyrum esculentum]. AB - The ultrastructures of storage protein accumulation in cotyledons and aleurone layer in buckwheat (Fagopyrum esculentum Monch) were investigated by electron microscopy. (1) On 15 days after anthesis (DAA), there are many dictyosomes and vacuoles accumulating protein in cytoplasm of the outer layer endosperm cells. These cells containing abundant aleurone grains (1-2 microm in diameter) from vacuoles accumulated storage protein form aleurone layer where division of aleurone grain is also observed on 25 DAA. (2) On 20 DAA, protein begins to accumulate in vacuoles of cotyledon cells. At the early stage of protein deposition, vacuoles are becoming smaller ones by ingrowth of tonoplast or by pinch-off. A multitude of ribosomes, many dictyosomes and electron-dense vesicles (0.1-0.7 microm in diameter) coated with membrane are observed in cytoplasm of cotyledon cells at every stage of protein accumulation. Originated from the separation of saccules containing protein at the fringe of dictyosome, these vesicles (0.1-0.2 microm in diameter) probably increase their volume by fusion each other. They play the leading role in transporting storage protein into vacuoles that become PSVs in cotyledon cells of buckwheat. On 25 DAA, there are many PSVs (1-3 microm in diameter) and some vesicles (0.1-0.7 microm) filled with protein in mature cotyledon cells. These vesicles may be also play a part in accumulating storage protein in mature buckwheat seed. PMID- 15346986 TI - [Long terminal repeats of retroposons in Candida albicans]. AB - 25 Candida albicans strains have been analysed on Southern hybridization patterns with alpha, beta, gamma, kappa probes. The alpha, beta, gamma, kappa, elements can be used to distinguish the differences and relations between C. albicans strains. Chromosomal localization analysis indicated that the alpha, beta, gamma, kappa, elements were located on several different chromosomes in Candida albicans strains. Northern hybridization showed that alpha, beta, gamma were transcribed actively and regulated by different stimuli. The kappa was inactive. Our results indicated that in Candida albicans there were many kinds of retroposons whose numbers and distribution were different. These retroposons could be used to identify Candida albicans strains as moleculer markers. PMID- 15346987 TI - [Expression activity of maize Ubi-1 promoter in fertile transgenic maize plants]. AB - The encoding sequence of gus gene from Escherichia coli was fused with maize Ubi 1 promoter and was introduced into maize genome via particle bombardment. Fertile transgenic maize plants were regenerated from bombarded type-I calluses which were derived from scutellar tissue of immature embryos based on PPT selection. Expression activity of gus gene under the control of Ubi-1 promoter was analysed using histochemical method, and the results showed that gus gene expressed in most tissues except anther. Ubi-GUS expression in pollen, egg cell and T1 immature embryos revealed that this promoter was active in early stages of plant development. Histochemically stained pollen grains of T0 plants showed a 1:1 segregation of the gus gene, which suggested that the foreign gene was inherited in Mendelian model in these plants. In addition, maize Ubi-1 promoter could reduce the copy number of foreign genes in transgenic maize plants, which might be useful in avoidance of gene silencing. T1 seeds have been harvested. PMID- 15346988 TI - [Construction of an angiostatin eukaryotic expression vector and characterization of its inhibitory efficiency in B16 melanoma bearing mice]. AB - Recent studies have demonstrated that angiostatin, a newly discovered specific inhibitor of endothelial cells, may significantly suppress the growth of a variety of tumors. We constructed an eukaryotic expression vector containing angiostatin (pAG3 ). To study the effect of pAG3, we intramuscularly injected pAG3 into B16 melanoma bearing C57 mice, we found that pAG3 could obviously inhibit tumor growth and reduce the size of tumors in B16 melanoma bearing C57 mice compared to the untreated control mice. In addition, we also investigated whether pre-treatment of pAG3 can prevent the tumor formation in mice treated with B16 melanoma. Normal C57 mice which received 5 days of treatment of pAG3 prior to implanting tumors resulted in the inhibitory effect when compared to control mice. No promotive effect was observed when pAG3 was combined with DTIC (Dacarbazine). These findings provide a basis for the further development of nonviral delivery of angiogenic gene therapy. PMID- 15346989 TI - [The morphology observation, mechanics intensity test and genetic analysis of brittle culm mutant bcm581-1 in rice]. AB - A rice brittle culm mutant bcm581-1 which derived from the Ds transposone transformation population was found, but the mutant was identified that it was not to be induced by Ds transposone insertion through PCR. The examination of the vascular bundle and cortical fibre cells in culm under the light and electron microscope showed that, the number of cortical vascular bundle of mutant was much more, the hollow among the cortical vascular bundle was deeper, and the cell walls of cortical fibre cells were thinner than the normal. The test of culm mechanics intensity showed that the load, elongation, strain, and stress of bcm581-1 were 5-9 times lower than normal. The moisture content and the wide fibre content of culm were test, the former was 3.5% higher, but the latter was 8.12% lower than normal. The analysis of genetic segregation in F2 and F1B1 population indicated that the brittle culm mutant was controlled by one recessive gene. PMID- 15346990 TI - [A comparatively histological observation on the megagametophytic abortion of female-sterile rice FS-1 and its maternal parent Fujisaka 5]. AB - A female-sterile rice FS-1, derived from the progeny of a backcross of F1 of Fujisaka 5 (Oryza sativa subsp. japonica, FJ) and Tjina (Oryza sativa subsp. indica) with FJ were used for histological study of megagametophytic abortion. FS 1 has a very low spikelet fertility (about 3%) and was regarded to have no true embryo sac. Paraffin sectioning was adopted to study the megagametophytic development of FS-1 compared with its maternal parent FJ. The materials were cultivated in field condition in Tsukuba, Japan and sampled at the middle stage of the panicle development. The results obtained were as follows: (1) FJ showed a normal megagametophytic development and resulted into a mature megagamete while most of the embryo sac of FS-1 underwent megaspore abortion. The remains of the embryo sac and the degenerated surrounding cells filled up the space where the normal embryo sac was expected to exist. (2) Abortion took place when the functional megaspore was being formed. Both the three nonfunctional megaspores near the micropyle and the functional megaspore farthest from the micropylar end were degenerated. No two-nucleus, four-nucleus and eight-nucleus embryo sac were found in the present study. It is concluded tentatively that the megagametophytic abortion of FS-1 occurred when the functional megaspore was being formed. PMID- 15346991 TI - [The identification of mouse cloned SFA DNA]. AB - For some basic investigation and the construction of artificial chromosomes, cloned centromeric DNAs identified on a firm ground are required. Thus, in the present work a preliminary screened clone of 13.5 kb DNA, 6# clone, form a mouse centromeric library contructed previously in our library was futher investigated by FISH and PCR. It was found that mouse 6# cloned SFA DNA, as shown by FISH is a fragment of mouse centromeric DNA. Evidence was also observed that 6# cloned SFA DNA consists of mouse minor satellite DNA and other DNA sequences. PMID- 15346992 TI - [Isolation of endophytic bacteria in potato and test of antagonistic action to bacterial ring rot of potato]. AB - In this study, two hundred and forty bacterial strains were isolated from inner tissue of potato tubers collected from DaTong, TaiYuan and Inner Mongolia Autonomous regions. On the basis of antagonistic examination in vitro, fifty and five bacteria strains were characterized for antagonistic bacteria to ring rot of potato. It was 22.9 percentage of all bacteria strains. The biggest radius of suppression circle was 13 mm. Nine strains were chosen for their suppression of bacterial ring rot, blackleg and dry rot of potato. These strains were bacteriologically ideatified. Strain 118 was Pseudomonas fluorescens biovar V. Strain 110 was Bacillus pumilus. Strain 085 was Bacillus stearothermophilus. Strain 069 was Erwinia herbicola. Strain 043 was Xanthomomas fragariae. Strain 116 was Curtobacterium. Strains A-10' and T3 were Bacillus. Strain H1-6 was Pseudomonas fluorescens. PMID- 15346993 TI - The elderly patient with cancer: new approaches for improved outcomes. PMID- 15346994 TI - Cancer: a disease of the elderly. AB - Healthcare providers can expect to see a steadily growing number of elderly patients with cancer. The US population is aging, and the incidence of cancer and cancer mortality are increasing. Sex-related differences in the incidence of cancer become apparent after the age of 64 years, with a higher lifetime risk of cancer in men. The process of aging at the subcellular level contributes to the development of cancer. The cost of cancer to society is hundreds of billions of dollars a year, and the direct, indirect, and intangible costs of cancer to a patient can be devastating. Elderly patients' out-of-pocket expenses for treatment and care can come at a time when their financial reserves are diminishing. Managing cancer in older patients is complicated by their shorter life expectancy and greater risk of treatment complications because of the high prevalence of comorbidities, decreased hematopoietic reserves, cognitive impairment, functional decline, and depression. The goals of managing cancer in older patients may differ from those in younger patients, as the potential benefits of treatment decrease and the risks of treatment and the need for supportive care increase. PMID- 15346995 TI - Poor prognosis in elderly patients with cancer: the role of bias and undertreatment. AB - Undertreatment in elderly patients with cancer may be a primary reason for their having poorer outcomes than younger patients. Several studies have shown, however, that otherwise-healthy elderly patients can obtain the same benefits from the same treatment as younger patients. Undertreatment because of age, independent of comorbidities, is apparent in studies of malignancies such as lung, colorectal, and breast carcinomas and lymphoma. The reasons for the bias may include patient preferences, physicians' tendencies to treat patients according to their chronologic age, and a lack of evidence-based guidelines for treating older patients. In many conditions there are few data on the efficacy of treatment in older patients, because of their underrepresentation in randomized controlled trials. Trials of cancer therapy that focus on, or even include, elderly patients should be a priority in the future. Elderly patients are perceived to be less able than younger patients to tolerate standard chemotherapy regimens because of their decreased hematopoietic reserves, the high prevalence of comorbidities, and the greater risk of toxicity. This belief leads to undertreatment through dose reductions and delays that may compromise outcomes. As an alternative, using prophylactic growth factors for hematopoietic support makes it possible for elderly patients to benefit from full doses of the chemotherapy. Establishing age-based standards for cancer care will become more important as the US population ages. PMID- 15346996 TI - Greater risks of chemotherapy toxicity in elderly patients with cancer. AB - Complications of cytotoxic chemotherapy are more common in older patients (65 years of age and older) with cancer than in younger patients, and the occurrence of myelosuppression, mucositis, cardiodepression, peripheral neuropathy, and central neurotoxicity can complicate treatment. Age-related physiologic changes that can increase the toxicity of chemotherapy are decreased stem-cell reserves, decreased ability to repair cell damage, progressive loss of body protein, and accumulation of body fat. A decline in organ function can alter the pharmacokinetics of many of the commonly used chemotherapeutic agents in some elderly patients, making toxicity less predictable. Comorbidities increase the risk of toxicity through their effects on the body. Furthermore, the drugs used to treat comorbidities may interact with chemotherapeutic drugs, potentially increasing toxicity in elderly patients. Prospective trials in older patients with lymphoma or solid tumors have found that age is a risk factor for chemotherapy-induced neutropenia and its complications. Anemia may be present because of the disease or its treatment, and, if left uncorrected, it can alter drug activity and increase toxicity. Being able to predict which elderly patients are at greater risk of toxicity on the basis of pretreatment factors would be valuable, and there is a need for prospective trials to determine regimen- and patient-specific prognostic factors. Effective management of the toxicity associated with chemotherapy with appropriate supportive care is crucial, especially in the elderly population, to give them the best chance of cure and survival, or to provide palliation. For example, management of neutropenic complications with colony-stimulating factors makes treatment with standard-dose chemotherapy possible, which can lead to better outcomes. A better understanding of drug activity and toxicity in older patients is necessary for developing guidelines for safe and effective treatment. Few randomized controlled trials of antitumor drugs in older patients with cancer have been conducted, but a number of agents with favorable efficacy and toxicity profiles in elderly patients have been identified. PMID- 15346997 TI - Family caregivers of elderly patients with cancer: understanding and minimizing the burden of care. AB - Family caregivers play an essential role, usually unpaid, in caring for patients with cancer. Most older patients with cancer are cared for by a family member, who may not be prepared for the challenges. The needs of older patients are diverse and may include assistance with medication, transportation for treatment, activities of daily living, and emotional support. The activities that caregivers find most stressful include helping patients with their self-care, managing their treatment and symptoms, and dealing with the suffering of a family member. Families may be affected by other stressors, such as changes in roles and employment and disruptions in schedules (eg, frequent clinic visits). Caregivers respond to these stressors differently; older spouses may be particularly vulnerable because of their own frailty. There can also be negative effects on caregivers' psychological, social, or physical health functioning. Social and economic deficits due to caregiving may include lifestyle disruption, less socializing, and greater out-of-pocket and lost productivity costs. Studies have shown, however, that caring for an older person with cancer also has rewards, such as satisfaction and a greater sense of self-worth. The negative aspects of caregiving can be lessened by psychological support and assistance in problem solving from healthcare professionals. Caregivers should also be provided with options to reduce the stress of frequent clinic visits, such as using long-acting growth factors or telephone triage. Educating caregivers on pertinent aspects of cancer management and the community resources available to them can be done through individual contact with health professionals or through more-formal educational programs. PMID- 15346998 TI - New paradigms for treating elderly patients with cancer: the comprehensive geriatric assessment and guidelines for supportive care. AB - Strategies for treating cancer are evolving to address the growing number of elderly patients with cancer. Older patients have highly variable physiologic ages, and their treatment should be individualized for optimal outcomes. Treatment paradigms should also take into account the diversity of patients' life expectancy, functional reserve, social support, and personal preference. A Comprehensive Geriatric Assessment (CGA) is a useful tool for estimating life expectancy and tolerance of treatment and for identifying reversible factors that may interfere with cancer treatment, including depression, malnutrition, anemia, neutropenia, and lack of caregiver support. Adopting a common language to describe older patients may facilitate the design and analysis of studies to determine effective drugs and care strategies for them. Information from a CGA can guide the prescription of potentially curative therapy, determine the best use of supportive care agents, and help identify frail patients for whom palliative care is the best option. There is evidence in a number of settings that the routine use of a CGA has a positive effect on health outcomes by reducing hospitalizations, preserving functional independence, and preventing geriatric syndromes. Guidelines for supportive care are also important in treating elderly patients with cancer. Pain, caused by cancer or its treatment, is prevalent, and guidelines for its assessment and treatment should be implemented to improve quality of life. Toxicities such as neutropenia and mucositis should be managed aggressively. Growth factors reduce the incidence and severity of neutropenia and its complications in older patients, particularly when they are administered in the early cycles of chemotherapy. The development of effective strategies for the management of toxicity caused by anticancer drugs may help the elderly, as much as younger patients, expect and look forward to a positive outcome with their treatment. PMID- 15346999 TI - Neurological complications of lumbar artificial disc replacement and comparison of clinical results with those related to lumbar arthrodesis in the literature: results of a multicenter, prospective, randomized investigational device exemption study of Charite intervertebral disc. Invited submission from the Joint Section Meeting on Disorders of the Spine and Peripheral Nerves, March 2004. AB - OBJECT: Arthrodesis is the gold standard for surgical treatment of lumbar degenerative disc disease (DDD). Solid fusion, however, can cause stress and increased motion in the segments adjacent to the fused level. This may initiate and/or accelerate the adjacent-segment disease process. Artificial discs are designed to restore and maintain normal motion of the lumbar intervertebral segment. Restoring and maintaining normal motion of the segment reduces stresses and loads on adjacent level segments. A US Food and Drug Administration Investigational Device Exemptions multicentered study of the Charite artificial disc was completed. The control group consisted of individuals who underwent anterior lumbar interbody fusion involving BAK cages and iliac crest bone graft. This is the first report of Class I data in which a lumbar artificial disc is compared with lumbar fusion. METHODS: Of 304 individuals enrolled in the study, 205 were randomized to the Charite disc-treated group and 99 to the BAK fusion treated (control) group. Neurological status was equivalent between the two groups at 6, 12, and 24 months postoperatively. The number of patients with major, minor, or other neurological complications was equivalent. There was a greater incidence of both major and minor complications in the BAK fusion group at 0 to 42 days postoperatively. Compared with data reported in the lumbar fusion literature, the Charite disc-treated patients had equivalent or better mean changes in visual analog scale and Oswestry Disability Index scores. CONCLUSIONS: The Charite artificial disc is safe and effective for the treatment of single level lumbar DDD, resulting in no higher incidence of neurological complications compared with BAK-assisted fusion and leading to equivalent or better outcomes compared with those obtained in the control group and those reported in the lumbar fusion literature. PMID- 15347000 TI - The future in the care of the cervical spine: interbody fusion and arthroplasty. Invited submission from the Joint Section Meeting on Disorders of the Spine and Peripheral Nerves, March 2004. AB - In the past 50 years tremendous advances have been made in the treatment of cervical disc disease with cervical fusion. Fusion rates have surpassed 95% after application of anterior cervical implants. Adjacent-segment degeneration, however, has plagued the long-term clinical success of cervical fusion. Cervical arthroplasty has been introduced to maintain cervical motion and potentially avoid or minimize adjacent-segment degeneration. If cervical arthroplasty is successful, the long-term results of surgery for cervical disc disease may improve; however, there are associated drawbacks that must be overcome. Implant wear, fatigue, and failure have been reported in cases of large-joint arthroplasty, and research is underway to limit these problems in cervical arthroplasty. In this article the authors trace the evolution of cervical fusion and the new technique of cervical arthroplasty. The nomenclature of cervical arthroplasty will also be introduced. PMID- 15347001 TI - Three-level and four-level anterior cervical discectomies and titanium cage augmented fusion with and without plate fixation. AB - OBJECT: Cage-assisted anterior cervical discectomy and fusion (ACDF) has proven to be a safe and effective procedure for the treatment of one- and two-level degenerative disc disease (DDD). To the authors' knowledge, clinical results after three- and four-level interbody cage-augmented ACDF have not been reported in the literature. The authors investigated the safety and effectiveness of titanium cages used in such procedures and evaluated the results in cases with or without plate fixation. METHODS: Fifty-six patients suffering from cervical DDD were divided into two groups. Group 1 included 32 patients who underwent titanium cage-assisted ACDF; Group 2 included 24 patients who underwent the same procedure, supplemented with plate fixation. The cervical DDD was confirmed by radiography and magnetic resonance imaging. The patients underwent radiographic evaluation to assess cervical lordosis, segmental height of cervical spine, the height of the foramina, and spinal stability. Neurological outcomes were assessed using the Japanese Orthopaedic Association (JOA) scores. Neck pain was graded using a 10-point visual analog scale (VAS). The follow-up period ranged from 13 to 28 months (mean 17.2 months). In both Groups 1 and 2 significant increase (p < 0.001) was demonstrated in the JOA scores (preoperatively 10.7 +/- 2.4 and 11.1 +/- 2, postoperatively 13.9 +/- 2.2 and 14.1 +/- 2.3, respectively) and VAS pain scores (preoperatively 8.8 +/- 0.9 and 8.5 +/- 1, postoperatively 3.1 +/- 2.1 and 2.8 +/- 1.8, respectively); however, there was no significant intergroup difference. A significant increase in the cervical lordosis, foraminal height, and segmental height was observed in both groups. Good stability of cage fusion was obtained in both groups 12 months postoperatively (90.6% in Group 1 and 91.7% in Group 2); however, there were no statistically significant intergroup differences. The complication rate in Group 2 was higher than that in Group 1. The hospital length of stay in Group 1 was significantly lower than in Group 2 (p < 0.001). CONCLUSIONS: Analysis of these findings demonstrated that titanium cage assisted ACDF provided long-term stabilization, increased lordosis, increased segmental height, and increased foraminal height. In both groups good neurological outcomes were achieved and donor site morbidity was avoided. The lower complication rate and shorter hospital stay, however, make the cage assisted fusion without plate fixation better than with plate fixation. PMID- 15347002 TI - Long-term results of expansive open-door laminoplasty for ossification of the posterior longitudinal ligament of the cervical spine. AB - OBJECT: Numerous surgical procedures have been developed for treatment of ossification of the posterior longitudinal ligament (OPLL) of the cervical spine, and these can be performed via three approaches: anterior, posterior, or combined anterior-posterior. The optimal approach in cases involving OPLL-induced cervical myelopathy, however, remains controversial. To address this issue, the authors assessed the benefits and limitations of expansive open-door laminoplasty for OPLL-related myelopathy by evaluating mid- and long-term clinical results. METHODS: Clinical results obtained in 72 patients who underwent expansive open door laminoplasty between 1983 and 1997 and who were followed for at least 5 years were assessed using the Japanese Orthopaedic Association (JOA) scoring system. The mean preoperative JOA score was 9.2 +/- 0.4; at 3 years postoperatively, the JOA score was 14.2 +/- 0.3 and the recovery rate (calculated using the Hirabayashi method) was 63.1 +/- 4.5%, both having reached their highest level. These favorable results were maintained up to 5 years after surgery. An increase in cervical myelopathy due to progression of the ossified ligament was observed in only two of 30 patients who could be followed for more than 10 years. Severe surgery-related complications were not observed. Preoperative JOA score, age at the time of surgery, and duration between onset of initial symptoms and surgery affected clinical results. CONCLUSIONS: Mid-term and long-term results of expansive open-door laminoplasty were satisfactory. Considering factors that affected surgical results, early surgery is recommended for OPLL of the cervical spine. PMID- 15347003 TI - Does chronic cervical myelopathy affect respiratory function? AB - OBJECT: Although respiratory function is often impaired by acute cervical spinal cord injury, changes in respiratory function in patients with chronic cervical myelopathy (CCM) are not well documented. The purpose of this study was to evaluate the respiratory function of patients with CCM. METHODS: Spirometric parameters were measured in 94 patients with CCM before they underwent expansive laminoplasty. These measurements were compared with those obtained in age- and sex-matched control group patients without myelopathy. The study patients were also subdivided into two groups: those with spinal compressive lesions above or below the C3-4 disc level were compared in terms of respiratory function. The vital capacity values measured in patients with CCM were significantly lower than those in the control group. In patients in whom spinal cord compression was present above C3-4, vital capacity values were lower than in patients in whom the compression level was below C3-4. The resting respiratory rate per minute was elevated in the CCM group. Peak expiratory flow rate was significantly decreased, and expiratory velocities at 50 and 25% of vital capacity were significantly increased in the CCM group. CONCLUSIONS: The results indicated that expiratory flow may be impaired or incomplete in patients with CCM. An underlying subclinical respiratory dysfunction appears to be associated with CCM. PMID- 15347004 TI - Cervical radiculopathy, entrapment neuropathy, and thoracic outlet syndrome: how to differentiate? Invited submission from the Joint Section Meeting on Disorders of the Spine and Peripheral Nerves, March 2004. AB - The common diagnoses of cervical radiculopathy and upper-extremity entrapment neuropathies can at times be difficult to differentiate. Additionally, thoracic outlet syndrome is often diagnosed when, in fact, the problem is radiculopathy or neuropathy. Another source of confusion, especially in older patients, is neuralgic amyotrophy, brachial plexitis, or the Parsonage-Turner syndrome. The differential diagnosis of unilateral arm pain, weakness, and/or sensory loss includes all of these problems. The clinical and electrodiagnostic features of each are discussed as an aid to distinguishing between these common and similar entities. PMID- 15347005 TI - Evaluation and treatment of congenital and developmental anomalies of the cervical spine. Invited submission from the Joint Section Meeting on Disorders of the Spine and Peripheral Nerves, March 2004. AB - Congenital and developmental osseous abnormalities of the cervical spine can result in neural compression ranging from the medulla oblongata to the cervicothoracic spinal cord junction. These may present in infancy as scoliosis and even limb weakness. A high index of suspicion is essential. Neurodiagnostic imaging relies on high-resolution computerized tomography (CT) scanning and three dimensional CT reconstructions as well as magnetic resonance imaging and angiography. The anatomical/physiological CT factors considered when developing a surgical approach are: 1) the stability and reducibility of the lesions; 2) direction and manner of encroachment of the lesion on the neural structures; 3) neural and vascular abnormalities; and 4) growth potential of the affected area. Primary stabilization is required for reducible lesions, whereas irreducible lesions are decompressed in the manner in which encroachment has occurred. Instability, whether present before or after operative intervention, required spinal stabilization. Illustrative examples of this approach are presented. PMID- 15347006 TI - Insertion torque and pullout force of rescue screws for anterior cervical plate fixation in a fatigued initial pilot hole. AB - OBJECT: The purpose of this study was to investigate whether thicker-core diameter screws increase fixation strength in the cervical spine. METHODS: Bone mineral density (BMD) was determined for each vertebral body (VB) obtained in six human C4-7 segments. Based on their BMD, the specimens were assigned to one of two groups in which torque and pullout force were tested. Two initial pilot holes were drilled into the VBs and tests were first performed using a standard screw. The test was repeated using a thicker rescue screw inserted into the same initial pilot hole. The mean value of peak torque and pullout force resulting from the single left/right measurements was used for statistical analysis. A t-test was performed to determine the effect of screw design on peak torque and pullout force. Moment correlation coefficients were calculated to determine the effect of BMD on peak torque and pullout force. Mean insertional peak torque for the standard screw was 82.1 N/cm and that for the rescue screw was 47.6 Ncm (p < 0.001). There was a strong correlation between insertional peak torque and BMD for both standard screws (r = 0.71, p = 0.02) and rescue screws (r = 0.59, p = 0.07). The mean pullout force for standard screws was 464.7 N, whereas it was 164.5 N for rescue screws (p < 0.001). There was a strong correlation between pullout force and BMD for both standard (r = 0.75, p = 0.0081) and rescue screws (r = 0.7, p = 0.025). CONCLUSIONS: Uncemented rescue screws that have been inserted into a fatigued hole in the cervical VB do not strengthen the screw-bone interface compared with the strength initially conferred by a standard screw. PMID- 15347007 TI - A comparison of simulator-tested and -retrieved cervical disc prostheses. Invited submission from the Joint Section Meeting on Disorders of the Spine and Peripheral Nerves, March 2004. AB - OBJECT: Total joint arthroplasties most commonly fail because the implant becomes worn and a host inflammatory response subsequently develops. Both the material response to the biological environment and the host response to the device must be thoroughly evaluated to establish the efficacy of cervical arthroplasty. Analyses of devices explanted in humans allow evaluation of both responses. Hypothetical wear rates can be determined by comparing in vivo wear with simulator-derived wear. The purpose of this study was to perform explant analyses involving the Bryan and Prestige discs and compare these results with those obtained using spine simulators. METHODS: Of the approximately 5500 patients treated with the Bryan disc, 11 have undergone explantation of the device. Six of these devices were analyzed for dimensional and chemical changes. Three of the approximately 300 implanted Prestige discs were retrieved, and two were examined microscopically. Histological specimens were assessed for wear particles and host inflammatory response. Additionally, the extent of simulator-produced wear was compared with that demonstrated in the retrieved specimens. CONCLUSIONS: The simulator-generated results predict adequate wear-related characteristics for both the Bryan and Prestige prostheses for a minimum of 40 years. Comparison of data with those of the retrieved specimens indicates that the wear was more minimal than predicted in simulators by five- to 10-fold. In no instance did the revisions result from failure of the device due to a reaction to wear debris, fracture, polymer oxidation, or metal corrosion. The inflammatory response seen in the periprosthetic tissues was minimal and not characteristic of inflammatory responses in failed diarthrodial joint arthroplasties. PMID- 15347008 TI - Vertebral synovial chondromatosis. Report of two cases and review of the literature. AB - Synovial chondromatosis is an uncommon disorder characterized by the formation of multiple cartilaginous nodules within the synovium, most commonly affecting large joints. Its involvement with the spine is rare; only six cases have been reported. The authors describe two patients with synovial chondromatosis involving the cervical spine. In the first case, synovial chondromatosis arose from the left C1-2 facet joint. This patient underwent a two-stage procedure including a posterior approach for tumor resection and occipitocervical fusion as well as a transmandibular circumglossal approach to the anterior craniocervical junction to complete the tumor removal. Interestingly, on histopathological examination, scattered foci of low-grade chondrosarcoma were intermixed within the synovial chondromatosis. To the authors' knowledge, this is the first report of secondary low-grade chondrosarcoma arising in vertebral synovial chondromatosis. In the second case, synovial chondromatosis involved the left C4 5 facet joint. Tumor resection and cervical fusion were performed via a posterior approach. In this report, the authors describe the clinical presentation, radiographic findings, operative details, histopathological features, and clinicoradiological follow-up data obtained in these two patients and review the literature pertaining to this rare entity. PMID- 15347009 TI - Positional vomiting due to a thoracic spinal dural arteriovenous fistula. Case report. AB - The authors report the unique case of a patient with a thoracic spinal dural arteriovenous fistula (DAVF) causing remote brainstem symptoms of positional vomiting and minimal vertigo. Magnetic resonance (MR) imaging of the brain demonstrated high signal abnormality in the medulla, presumably related to venous hypertension, and spinal MR imaging revealed markedly dilated veins along the dorsal aspect of the cord. Spinal angiography confirmed the presence of a thoracic spinal DAVF. Disconnection of the DAVF from the spine resulted in a marked improvement in symptoms and resolution of the preoperative MR imaging documented abnormalities. The authors highlight the rare syndrome of positional vomiting as a brainstem symptom and conclude that spinal DAVFs should be considered in the differential diagnosis of high signal MR imaging abnormalities localized to the brainstem. PMID- 15347010 TI - Transdural herniation of the thoracic spinal cord: untethering via a posterolateral transpedicular approach. Report of three cases. AB - Anterior spinal cord herniation is a well-documented condition in which the thoracic cord becomes tethered within a defect in the anterior dura mater. Typical procedures have involved a posterior approach with direct manipulation of the thoracic cord to expose and blindly release its point of tethering. The authors report three cases in which a novel approach for the treatment of anterior thoracic cord herniation was performed, cord manipulation and traction are minimized, and direct dural repair of the defect is performed. PMID- 15347011 TI - Percutaneous vertebroplasty for the treatment of burst fractures. Case report. AB - Percutaneous vertebroplasty is a useful procedure for patients with vertebral osteoporotic compression fractures; however, there has been no mention in the literature of the use of percutaneous vertebroplasty for the treatment of traumatic spinal fractures. The authors report the case of a 33-year-old man who harbored L-1, L-2, and L-5 burst fractures sustained in a work-related accident. The patient was successfully treated by percutaneous vertebroplasty with polymethylmethacrylate. The authors propose this procedure as a useful intervention in selected patients with lumbar burst fractures. The complications associated with major surgical procedures are absent. PMID- 15347012 TI - Spontaneous spinal hematomas and low-molecular-weight heparin. Report of four cases and review of the literature. AB - The purpose of this article is to raise awareness of spontaneous spinal hematomas that develop after administration of low-molecular-weight heparin therapy. The authors describe four patients in whom these hematomas developed without precipitating events while receiving a treatment dose of enoxaparin (Clexane) (approximately 1 mg/kg). Spontaneous spinal hematomas (not related to trauma, surgery, or lumbar puncture) are a rare clinical entity. Several causes have been identified, including acquired and congenital clotting abnormalities and underlying vascular lesions. Aspirin, warfarin, tissue plasminogen activator, and heparin have all been implicated in causing spinal hematomas. Concerns regarding the use of low-molecular-weight heparin agents in neuraxis anesthesia have been well documented. Their possible contribution to nontraumatic spinal hematomas has been less well described. The authors believe that low-molecular-weight heparin agents present a small but significant risk of spinal hematoma. This should be considered when prescribing therapy because such a complication may be catastrophic. PMID- 15347013 TI - Multiple vertebral fractures in osteogenesis imperfecta treated by vertebroplasty. Case illustration. PMID- 15347014 TI - Paleopathological examination of medieval spines with exceptional thoracic kyphosis most likely secondary to spinal tuberculosis. Historical vignette. AB - Spinal tuberculosis (TB) infrequently involves more than one to three vertebrae, and kyphotic angulation of greater than 30 degrees is rare in paleopathological specimens and clinical studies. The authors describe findings obtained in two spines (dating from the Early and Late Middle Ages). Spinal TB was present in both as well as sharply angulated kyphosis (approximately 180 degrees) resulting from complete destruction of five and seven vertebral bodies, respectively. In these two specimens obtained in individuals older than 12 years of age at the time of death we observed no involvement of the disease in posterior vertebral elements, and the laminae showed osseous fusion without signs of infection. The osseous diameter of the spinal canal was not narrowed in either case. These findings are discussed in the context of modern medical knowledge and paleopathological and genetic examinations. PMID- 15347015 TI - A different tack to an old challenge. Does new managed care entrant have the answer for managing behavioral health? PMID- 15347016 TI - How to accomplish practice change in behavioral healthcare in less than one year. AB - This is a challenging time in the field of public mental health. State and federal funding for public behavioral healthcare is declining in a time when the demand for services is increasing. Philosophical changes in mental healthcare service delivery, underscored by the tenets from the President's New Freedom Commission on Mental Health, are creating additional challenges for practitioners. The Adult Network of Pikes Peak Mental Health (PPMH) has developed a model for providing behavioral healthcare that meets these challenges while providing greater access to care, holistic care delivered in natural settings and, at the same time, maintaining the quality clinical outcomes at reduced costs. PMID- 15347017 TI - Field leaders move forward on crafting a public campaign. PMID- 15347018 TI - Jail diversion program shows power of teamwork. PMID- 15347019 TI - Made in Arizona: a public sector, full-risk behavioral health system that works. PMID- 15347020 TI - Dilemmas in diagnosing and treating co-occurring disorders: an addiction professional's perspective. PMID- 15347021 TI - Psychiatrist parlayed $800 into model program for homeless. PMID- 15347022 TI - Multivariate discrimination among cryptic species of the mite genus Chaetodactylus (Acari: Chaetodactylidae) associated with bees of the genus Lithurgus (Hymenoptera: Megachilidae) in North America. AB - Twenty-seven morphological characters from 111 heteromorphic deutonymphs of the mite genus Chaetodactylus Rondani (Acari: Chaetodactylidae) were analyzed. The mites were collected from four species of bees of the genus Lithurgus Berthold (Hymenoptera: Megachilidae) in continental North America. Principal component and canonical variates analyses on Darroch and Mosimann shape and size-and-shape variables revealed the presence of three cryptic species. Chaetodactylus gibbosi sp. n. (Florida) is geographically isolated from C. lithurgi sp. n. distributed in Texas, New Mexico, Arizona, Colorado, and Idaho. Sympatric C. lithurgi and C. abditus sp. n. (USA: Arizona, Mexico: Socorro Is.) are seasonally isolated in Arizona. Chaetodactylus gibbosi is associated with a single bee species, Lithurgus gibbosus Smith in Florida. The host range of C. lithurgi includes bees flying predominantly in the spring: L. apicalis Cresson, L. littoralis Cockerell, and western L. gibbosus. Chaetodactylus abditus sp. n. is associated with L. planifrons Friese and L. echinocacti Cockerell, flying predominantly in the fall in Arizona. No distinct groups separated by geographic locality or size were detected in any species. A six-variable model developed by the canonical variates analysis and estimated using jackknife resampling and external validation (n = 100) is capable of classifying the three species with 100% accuracy. Factors that influenced speciation of cryptic species of Chaetodactylus associated with Lithurgus are discussed. Based on morphological and geographical data and data on mite associates, the western and eastern populations of the bee L. gibbosus are distinct. Therefore, the taxonomic status of L. gibbosus s. lat. should be reevaluated. PMID- 15347024 TI - Side effects of mancozeb on Typhlodromus pyri (Acari: Phytoseiidae) in vineyards: results of multi-year field trials and a laboratory study. AB - The side effects of mancozeb on the predatory mite Typhlodromus pyri were studied in 4-year field trials on grapevine and in the laboratory. In the field, the effect of mancozeb varied according to previous mancozeb use. In vineyards where mancozeb had commonly been used over years, this fungicide is generally slightly toxic, in some cases moderately toxic and rarely toxic. In plots were mancozeb has never been used, its effect on T. pyri was more pronounced and varied from moderately toxic to toxic. Despite the toxicity of mancozeb, T. pyri populations have never been eradicated. Laboratory results obtained with the French CEB guideline no. 167 confirmed those of the 4-year field study: mancozeb was significantly more toxic to T. pyri populations collected in plots where it had rarely been used before the field experiment. In plots where mancozeb had been used for a long time, the susceptibility of T. pyri populations to this fungicide was reduced and female survival, fecundity but also viability of female progeny were less affected by mancozeb. Even though toxicity of mancozeb increased in controlled conditions, a significant correlation was established between field and laboratory results. PMID- 15347023 TI - Molecular phylogeny of oribatid mites (Oribatida, Acari): evidence for multiple radiations of parthenogenetic lineages. AB - Nucleotide sequences of the D3 expansion segment and its flanking regions of the 28S rDNA gene were used to evaluate phylogenetic relationships among representative sexual and asexual oribatid mites (Oribatida, Acariformes). The aim of this study was to investigate the hypothesis that oribatid mites consist of species-rich clusters of asexual species that may have radiated while being parthenogenetic. Furthermore, the systematic position of the astigmate mites (Astigmata, Acariformes) which have been hypothesised to represent a paedomorphic lineage within the oribatid mites, is investigated. This is the first phylogenetic tree for oribatid mites s.l. (incl. Astigmata) based on nucleotide sequences. Intraspecific genetic variation in the D3 region was very low, confirming the hypothesis that this region is a good species marker. Results from neighbour joining (NJ) and maximum parsimony (MP) algorithms indicate that several species-rich parthenogenetic groups like Camisiidae, Nanhermanniidae and Malaconothridae are monophyletic, consistent with the hypothesis that some oribatid mite groups diversified despite being parthenogenetic. The MP and maximum likelihood (ML) method indicated that the D3 region is a good tool for elucidating the relationship of oribatid mite species on a small scale(genera, families) but is not reliable for large-scale taxonomy, because branches from the NJ algorithm collapsed in the MP and ML tree. In all trees calculated by different algorithms the Astigmata clustered within the oribatid mites, as proposed earlier. PMID- 15347025 TI - Habitat-related variation in infestation of lizards and rodents with Ixodes ticks in dense woodlands in Mendocino County, California. AB - During the spring and early summer of 2002, we examined the relative importance of Borrelia-refractory lizards (Sceloporus occidentalis, Elgaria spp.) versus potential Borrelia burgdorferi sensu lato (s.l.)-reservoirs (rodents) as hosts for Ixodes pacificus immatures in 14 woodland areas (six oak, five mixed oak/Douglas fir, and three redwood/tanoak areas) distributed throughout Mendocino County, California. Lizards were estimated to serve as hosts for 93-98% of all larvae and > or =99.6% of all nymphs infesting lizards or rodents in oak woodlands and oak/Douglas fir sites in the southern part of the county. In redwood/tanoak woodlands and oak/Douglas fir sites in northern Mendocino County, the contribution of rodents to larval feedings reached 36-69% but lizards still accounted for 94-100% of nymphal bloodmeals. From late April to mid-June, I. pacificus larvae were recovered from 95 to 96% of lizards and dusky-footed woodrats (Neotoma fuscipes) and from 59% of Peromyscus spp. mice. In contrast, 99% of lizards but few woodrats (15%) and none of the mice were infested by nymphs. Comparisons of tick loads for 19 lizard-Peromyscus spp. mouse pairings, where the lizard and mouse were captured within 10m of each other, revealed that the lizards harbored 36 times more larvae and >190 times more nymphs than the mice. In oak woodlands, loads of I. pacificus larvae decreased from late April/early May to late June for S. occidentalis lizards but increased for Peromyscus spp. mice. We conclude that the relative utilization of Borrelia refractory lizards, as compared to rodents, by I. pacificus larvae was far higher in dry oak woodlands than in moister habitats such as redwood/tanoak and oak/Douglas fir woodlands in northern Mendocino County. Non-lizard-infesting potential enzootic vectors of B. burgdorferi s.l. (I. angustus and I. spinipalpis) were recorded from rodents in three of six oak woodland areas, two of five oak/Douglas fir woodland areas, and two of three redwood/tanoak woodland areas. PMID- 15347026 TI - Water relations in eggs of the lone star tick, Amblyomma americanum, with experimental work on the capacity for water vapor absorption. AB - This study shows that water stress is not countered in eggs of the lone star tick, Amblyomma americanum (L.), using water vapor, and suggests involvement of liquid water as a developmental cue. Eggs fail to maintain an equilibrium water content in subsaturated air, hence, gain not equal to loss, with net water losses occurring at relative humidities near saturation and these eggs exhibit a three fold drop in viability, but not incubation period, as compared to eggs held in saturated air. Amblyomma americanum eggs are stenohydric and feature low 58% water content, slow water losses <1%/h, and an impermeable chorion wherein the Arrhenius activation energy, Ea = -66J/K, is suppressed. Thus, enhancement of water retention, not water vapor absorption, permits eggs to resist desiccation. PMID- 15347027 TI - [Problems and perspectives of preventive angioneurology]. PMID- 15347028 TI - [Characteristics of vertebral and muscular tonic syndromes in acute and remote periods of cervical whiplash injuries]. AB - The study elicited the peculiarities of vertebral and muscular tonic syndromes in acute and remote periods of whip cervical trauma (WCT). Forty patients in acute period of WCT (2nd-3rd degree of severity) and 30 patients in remote period of WCT, who experienced pain and other symptoms 6 months after the trauma (late whip syndrome--LWS) were examined. The control group included 30 patients with neck and arm pain due to cervical osteochondrosis. In WCT, comparing to cervical osteochondrosis, more marked movement restriction in sagittal plane, more frequent blockade of the lower cervical spine segments, stronger correlation between pain syndrome and movement restriction in the cervical segments, more frequent muscular tonic syndrome in the anterior neck muscles and deeper neck flexors were found. In LWS, in contrast to the acute period of WCT, dissociation between more restricted active and more preserved passive movements in the cervical segments, weaker correlation between emerging of pain syndrome and restriction of movement volume, more frequent blockade of the upper cervical segments, more frequent occurrence of supraspinal muscles and shoulder-scapular syndromes were detected. The data obtained revealed a complex mechanism of symptoms formation in WCT that should be taken into account in treatment planning for acute and remote periods of cervical trauma. PMID- 15347029 TI - [Typology of disordered eating behaviour (anorexia nervosa with bulimia and vomitomanic disturbances) in children and adolescents]. AB - Eighty children (66 girls and 14 boys), aged 6-16 years, with syndrome of anorexia nervosa (AN) have been studied. Two nosologic groups: pathological age crisis of childhood and adolescence (F50.0-F50.2 in ICD-10)--30 patients and slow progressive schizophrenia (schizotypical disorder, F21.3-F21.4)--50 patients are determined. The latter group was divided according to syndrome types: syndrome of AN (28 patients); AN syndrome with predominant bulimia (11 patients); AN syndrome with dominating bulimia and vomitomanic disturbances (11 patients). Syndrome structure was determined by the stages of the course, syntropy character with other psychopathological disorders and process of their reduction. Psychotic disorders in patients of the third type were most pronounced. In each group, statistically significant EEG features, comparing to control age-matched group, were revealed. The severer were psychopathological symptoms, the less was an amount of organized and the more--desynchronized "flat" EEG. PMID- 15347030 TI - [Behavioural disorders in children with slow-progressive schizophrenia]. AB - Sixty-five patients, aged 7-15 years, with established diagnosis of slow progressive schizophrenia (schizotypical disorder according to ICD-10) were studied. Behavioural disorders predominate in the clinical picture of the disease. Based on psychopathological analysis and taking into account the illness course and patient's age (ontogenetic maturity of psychic processes) and syndrome comorbidity, 3 types of behavioral disorders were distinguished: affective, overvalued and deficient. These types enables to predict disease course and a level of development of negative changes, to differentiate therapeutic policy and to evaluate adequately the adaptive reserves of the patients. PMID- 15347031 TI - [Use of ketogenic diet in treatment of pharmacoresistant epilepsy]. AB - Ketogenic diet (KD) is one of the most effective methods of epilepsy non-drug therapy. Ten patients with pharmacoresistant epilepsy, aged 1.5-18 years, were treated with individually matched and strictly controlled KD with high fat content and restricted amount of proteins and carbohydrates. After 1 year, only 4 patients kept the diet. Factors of efficacy, causes of withdrawal and side effects of KD are discussed. PMID- 15347032 TI - [Topamax monotherapy of partial epilepsy]. AB - Thirty patients diagnosed with partial epilepsy (PE): 17 cases with cryptogenic PE, 11--with symptomatic PE and 2--with symptomatic generalized PE, have been switched to topamax monotherapy, in dosage 50-200 mg daily, for 3 months. A type of seizures--simple partial, complex partial and secondary generalized, was taken into account, when considering topamax efficacy. A complete elimination of seizures was achieved after 3 months treatment in 80% of the cases, the mostly pronounced effect being demonstrated for secondary generalized seizures. A decrease of seizures frequency was shown for all types, but in simple partial and secondary generalized seizures the effect emerged earlier (after 1 month of the treatment) comparing to complex partial seizures. Good tolerability of topamax was observed. PMID- 15347033 TI - [Betaserc treatment of psychogenic vertigo]. AB - An effect of betaserc on the character and intensity of vertigo (V), psycho autonomic state of patients, vestibular functions and oculomotor system, using computer stimulating programs, has been studied in 39 patients with V. After 4 weeks of the treatment, betaserc exerted a positive effect on patients; however, the optimal effect was shown in patients with psychogenic V (73%). In this group, betaserc reduced both V and other psycho-autonomic disturbances. After betaserc treatment, patients with psychogenic V demonstrated a normalization of the vestibular responsiveness parameters in 70% of the cases and of the parameters of spontaneous oculomotor activity and pursuit eye function in 40 to 90%. PMID- 15347034 TI - [New data on sensibilizing therapy of alcoholic dependence]. AB - Lidevine was used in the treatment of 29 patients with chronic alcoholism with unsatisfactory results of previous therapy. The drug was included in the complex therapy at the stage of developing therapeutic remission and during a long-term (1 year) antirelapse treatment. Remission of 1-year duration was achieved in 12 (41%) patients. Use of lidevine contributes to remission stabilization and prevents development of alcoholism relapse after a single "break down". The importance of lidevine assignment in combination with psychopharmacological drugs reducing a drive for alcohol (antidepressants, anticonvulsants-normotimics, neuroleptics with sedative action, etc.), and a need of continuous control of the relatives of the patient over the intake of sensibilazing drug are proved. Good tolerance to the medication is shown. PMID- 15347035 TI - [The efficacy of calcium channel blockers in the treatment of affective disorders and pathologic drive for alcohol in patients with alcoholism in the remission period]. AB - Forty three patients with chronic alcoholism of stage II with secondary affective disorders (anxiety, subdepression) and actualization of pathological drive for alcohol in remission period were divided into 3 groups: patients were treated with nimodipine (90 mg/day during 10 days), patients were treated with nifedipine (45 mg/day during 10 days) and patients of control group (they received a placebo for 10 days). The study was double blind. The results revealed that both calcium channels antagonists reduced a level of depression measured with the Hamilton scale and Zung test. Ten-day nimodipine (but not nifedipine) course also caused a significant decrease of anxiety evaluated by the Spilberger test. Both medications, being significantly more efficient comparing to placebo, led to desactualization of pathological drive for alcohol in parallel with affective disorders reduction. Because of the absence of the depriming effect of these compounds on CNS as well as of dependence phenomenon, calcium channels antagonists may be used in narcology for stopping secondary affective disorders and actualization of pathological drive for alcohol in remission period. PMID- 15347036 TI - [Effect of cerebrolysin on the electroencephalographic indices of brain activity in Parkinson's disease]. AB - Cerebrolysin is a brain-derived peptide drug that increases the BBB-GLUT1 and MAP2 genes expression, thus exerting a neuroprotective effect. The present study aimed at investigating in patients with Parkinson's disease (PD) influence of Cerebrolysin infusions (intravenously, 10 ml during 10 days) combined with levodopa treatment on the electroencephalographic (EEG) indices of brain activity: P300 potential, contingent negative variation (CNV) and recovery functions of the cortical auditory evoked potentials, which reflect the postexcitatory inhibition at the paired stimulation. Nineteen PD patients, mean age 61.4 +/- 1.7 years; disease stage according to M.M. Hoehn and M.D. Yahr, 1967 2.2 +/- 0.1) and 18 age-matched healthy controls were studied. In the patients with essential differences of the EEG indices, comparing to the normal values, statistically significant changes were revealed: a decrease of P300 latency from 419.4 +/- 23.5 to 356.3 +/- 18.4 ms (8 patients, 42%); an increase of CNV duration from 423.1 +/- 93.3 to 600.6 +/- 38.5 ms; 2-fold increase of CNV mean amplitude and 3-fold increase of CNV square (8 patients, 42%) and strengthening of postexcitatory inhibition in auditory system at the paired stimulation (13 patients, 68%). In conclusion, Cerebrolysin may be recommended as an additional neuroprotective drug for brain functions improvement in the complex pathogenetic therapy of earlier PD stages. PMID- 15347037 TI - [Use of cluster analysis for the study of self-stigmatization phenomenon]. AB - Cluster analysis was used in the study of self-stigmatization phenomenon measured with the original self-stigmatization questionnaire in 81 patients with schizophrenia, affective and neurotic disorders. The answers presented by digital codes were analyzed statistically. Six factors of self-stigmatization were as follows: "a victim of stigma", "deidentification from the healthy", "predisposition to identification with mentally ill", "avoidance of mentally ill", "excuse by mental illness" and "generalization of stigma". PMID- 15347038 TI - [A comparative evaluation of immunomodulating drugs in the treatment of remitting multiple sclerosis]. PMID- 15347039 TI - [The phenomenon of "forced normalization" in the framework of psychotic changes in epileptic patients]. PMID- 15347040 TI - [Rational approach to peripheral arterial occlusive disease. Correcting pressure properly]. PMID- 15347041 TI - [High morbidity rate, therefore: peripheral arterial disease screening from age 45 on! (interview by Dr. Ulrike Wepner)]. PMID- 15347042 TI - [A warming by the German Cancer Society. Risk of breast cancer caused by deoderant sprays containing paraben? (interview by Christine Vetter)]. PMID- 15347043 TI - [Child with severe burns. First aid from the water faucet]. PMID- 15347044 TI - [Quality control with bite. What cardiologists can learn from McDonald's]. PMID- 15347045 TI - [Questions about Echinococcus multilocularis. Transmission by fallen fruit?]. PMID- 15347046 TI - [Treatment of allergic rhinitis]. AB - The allergic rhinitis is the most common allergic condition. The causes of allergic rhinitis are still much debated, but there is now a consensus on the need to initiate treatment as early as possible in order to avoid the spreading of the infection to the bronchi. In addition to allergy prevention, the aim of treatment is medical control of the allergic inflammatory reaction and thus improvement of the clinical symptoms. Depending on the severity of the disease and the predominant symptoms, this can be achieved with mast cell stabilizers, local and systemic H1 antagonists, local steroids and anticholinergic drugs. The WHO also recommends hyposensitization, which may be subcutaneous (SIT) or sublingual (SLIT). PMID- 15347047 TI - [Might it be allergic bronchial asthma?]. AB - The prevalence of allergy and asthma in our population is on the increase, as also, it is presumed, is the number of unrecognized cases. In the diagnostic evaluation of patients with respiratory problems, therefore, the question "might it be allergic bronchial asthma?" should be asked more frequently. The central diagnostic mainstays are the finding of reversible bronchial obstruction on pulmonary function testing, and the differential allergologic investigation. Once the diagnosis has been established, comprehensive management comprising pharmacotherapy, training and preventive measures can make it possible for the patient to enjoy a largely symptom-free life. The hope for the future is that an ever deeper understanding of the pathogenesis of asthma will enable effective preventive measures to be taken in childhood. PMID- 15347048 TI - [Allergic conditions of the eye]. AB - In the doctor's office, seasonal and non-seasonal conjunctivitis must be differentiated from more serious conditions. These include vernal conjunctivitis which, when chronic, represents a risk for corneal complications. In atopic keratoconjunctivitis, too, which manifests in every fourth patient with atopic dermatitis, the patient's vision is in danger. Furthermore, wearers of contact lenses may develop characteristic conjunctival changes or a contact allergy triggered by lens cleansing fluid. When the diagnosis has been established, treatment with local or systemic antihistaminics, H1 blockers or mast cell stabilizers can be initiated. In contrast, the indication for glucocorticoids should be established only by an ophthalmogist. PMID- 15347049 TI - [The 308 nm excimer laser--an advance in the treatment of psoriasis]. PMID- 15347050 TI - [Primary combination treatment of hypertension. Strategies for improved therapy outcomes]. PMID- 15347051 TI - [New evidence based medicine starting in the second quarter: death knoll for larger practices?]. PMID- 15347052 TI - [Co-pay reduced--free cards for doctor hopping patients]. PMID- 15347053 TI - [Consequences of general practice tax--point value far more than 5 cents?]. PMID- 15347055 TI - [Social security recipients: must they pay now or not?]. PMID- 15347056 TI - [Successful phase III study with serotonin-noradrenaline reuptake inhibitor. Soon a drug to control stress incontinence]. PMID- 15347057 TI - [Brief antibiotic therapy. 3 swallows in suppurative bronchitis]. PMID- 15347058 TI - [Previously life threatening attacks, now physically active without fear. Sports as asthma therapy]. PMID- 15347059 TI - [Use of triptanes according to indications. Risk of infarct is not increased]. PMID- 15347060 TI - [When compliance doesn't work out. Calculate the risk of stroke for hypertensive patients!]. PMID- 15347061 TI - [A compensated type 2 diabetic patient? Fasting blood glucose can be deceptive]. PMID- 15347062 TI - [Data comparison of osteoporosis patients demonstrates nasal administration is not optimal]. PMID- 15347063 TI - [Diagnostic quiz: back pain in malnutrition]. PMID- 15347064 TI - [Early diagnosis prevents long-term odyssey through specialty practices]. PMID- 15347065 TI - [10 minute consultation: chronic fatigue]. PMID- 15347066 TI - [Diagnosis at a glance: cafe au lait spots of the back and "freckles" in the axilla]. PMID- 15347067 TI - [Food intolerance or food allergy? Search for "culinary risk factors"]. PMID- 15347068 TI - [Search for the etiology in chronic diarrhea. Not every colitis is imaged with the endoscope]. PMID- 15347069 TI - [Palliative medicine: how can the family physician help, when recovery is not possible? (interview by Christine Vetter)]. PMID- 15347070 TI - [Penalty for relationships. Is marriage proscription near for physicians? ]. PMID- 15347071 TI - [Roentgen diagnosis responsible for cancer fatalities? Please no risk without benefit analysis!]. PMID- 15347072 TI - [Parkinson's disease--new horizons]. AB - New information about the pathology and genetics of Parkinson's disease opens up new possibilities for treatment. A key role is played by a pathologically folded protein, the distribution of which in the brain permits new conclusions about the stage of the disease. Gene mutations in affected families provide further clues as to the pathological process. In addition to symptom control, future treatment strategies will be aimed in particular at slowing down the underlying pathological process, and will therefore go in the direction of neuroprotection and neurorestoration. Dopamine agonists that can slow cell degeneration are already the first-line treatment in patients under 70 years of age. PMID- 15347073 TI - [Radionuclide techniques in the diagnosis of parkinsonian syndromes]. AB - The differential diagnosis of most parkinsonian syndromes can be adequately established on the basis of clinical criteria alone. The characterization of the dopaminergic system using nuclear imaging techniques such as PET and SPECT, is gaining in importance, in particular for the diagnosis of clinically unclear cases. For the first time, these techniques permit the investigation of the presynaptic and postsynaptic dopaminergic system in vivo, and thus the establishment of the differential diagnosis of parkinsonian syndromes at an early stage. Furthermore, the severity and progression of the disease can be assessed and the effect of putative neuroprotective drugs can be documented. PMID- 15347074 TI - [Neurosurgical treatment of Parkinson's disease]. PMID- 15347075 TI - [Water alone is not enough. Proper cleansing is the basis of acne treatment]. PMID- 15347076 TI - [Differential diagnosis of diseases of the external ear. What do red ears conceal?]. PMID- 15347077 TI - [Difficulties in asthma therapy]. AB - Asthma is the most frequent chronic disease in childhood with rising prevalance worldwide. Early antiinflammatory therapy can prevent irreversible damage to the lung. Effective drugs such as nedrocromil, DNCG, leukotrienes antagonists or inhaled corticosteroids are available and recommended. But they are not used as assigned. Guidelines may help to find the right medication but they have to be adapted to the individual and his situation. PMID- 15347078 TI - [Burns caused by acids and caustic agents]. PMID- 15347079 TI - [Chaos about general practice tax continues. Traveling to Absurdistan....]. PMID- 15347080 TI - [Representative survey reveals sequelae of reform. Every 4th will cut back on doctor visits]. PMID- 15347081 TI - [Gigantic study planned. 100,000 patients with COPD test general practice antibiotic]. PMID- 15347082 TI - [Capsule endoscopy study of gastrointestinal tolerance. Coxibes superior to NSAID plus antacid]. PMID- 15347083 TI - [HbA1c is too high despite tablets. With single injections toward better blood glucose values]. PMID- 15347084 TI - [Blocking cannabinoid receptors. Simultaneous control of overweight and nicotine addiction]. PMID- 15347085 TI - [Unfairly implicated as cost drivers. New drugs without a chance?]. PMID- 15347086 TI - [Especially in patients with diabetes, onychomycosis is not a harmless illness]. PMID- 15347087 TI - [Corticoid dose aerosol without FCKW. New technology assures optimal particle size]. PMID- 15347088 TI - [Delaying degeneration or promoting regeneration. 2 successful strategies in osteoporosis]. PMID- 15347089 TI - [The skin as chief manifestation. Is it really an allergy?]. PMID- 15347090 TI - [How does it work? Magnetic resonance cholangiopancreatography (MRCP)]. PMID- 15347091 TI - [Contribution of "Physicians for the 3rd Word" in Calcutta. Consultation on the refuse dump]. PMID- 15347092 TI - [Diagnosis at a glance: inflammatory boil at the elbow]. PMID- 15347093 TI - The lost innocence of research. PMID- 15347094 TI - Report on Ethics Summit. Initiative of oral health organizations. PMID- 15347095 TI - Lying through your teeth. AB - Evidence is presented that the concerns over diminished truth-telling in dentistry are a reflection of changes in society generally. Some of the reasons for dimmed truth-telling are listed, leading to a conclusion that society is in the process of rejecting an "objective" view of truth based on the word of authorities. An alternative, authentic conception of truth is proposed: truth is agreement that it is mutually advantageous for the parties involved to believe the promises they have made as a foundation for common action. Some of the characteristics of this view are discussed. PMID- 15347096 TI - Some observations on truth claims in a profession. PMID- 15347097 TI - Inside, outside, truth side up. PMID- 15347098 TI - Ethics fitness. PMID- 15347099 TI - Why we can't believe what we read. AB - It is argued that many claims made in the dental literature lack scientific grounding. Rather than become cynical, dentists are urged to use their own critical judgment and caution when reading the literature, especially articles and advertisements in the rapidly expanding area of dental materials. An example involving research on "condensable" resin-bonded composites is analyzed in detail, showing how an apparently credible claim can be lacking in support. PMID- 15347100 TI - The role of the Federal Trade Commission in advertising health products and services. AB - The Federal Trade Commission plays a unique role in enforcing well-established standards ensuring that consumers can make informed purchase and use decisions about health-related products and services based on truthful, non-misleading advertising claims while encouraging competition. Deceptive and unfair practices are defined. The importance of the "net impression" that ads convey to consumers and the need for substantiation of objective, factual claims is explained. The FTC uses its enforcement powers and consumer and industry outreach to create a climate for preventing misleading advertising. PMID- 15347101 TI - Will dentistry be left behind at the healthcare station? AB - The emerging opportunities for biological approaches to managing oral health were clearly apparent fifteen years ago. Responses in this direction have been tardy because of the economic prosperity of dental practice and the financial constraints on dental education. Representative existing and promising biology based technology is described. It is argued that education must be changed to teach the context and understanding of this technology. The alternative will be to concede important aspects of oral health care to physicians and others trained in their use. PMID- 15347102 TI - Research of importance to the practicing dentist: 2004 ADA research agenda. PMID- 15347103 TI - A call for ethics committees in dental organizations and in dental education. AB - It is argued that dental organizations need ethics committees to address growing concerns among the public regarding ethical conduct. Such committees could provide education, help formulate policy and guidelines, and develop case review and consultation, as well as create useful networks. The results of a survey of Canadian faculties of dentistry regarding ethics resources are presented. PMID- 15347104 TI - The second-best solution: an apology for democracy. AB - The best ways to make group decisions are by consensus or the use of rational rules. Unfortunately, these methods are not general and are limited to various special circumstances. Some poor alternatives include authoritarian rule or chaos. Democracy, essentially one voice and vote for each concerned individual, is the most practical general alternative. A catalogue of democracy's flaws is reviewed, but it is concluded that, faults and all, democracy remains the best choice. PMID- 15347105 TI - The malpractice crisis will cause a fundamental change in surgical practice. PMID- 15347106 TI - Percutaneous biopsy in evaluation of lung nodules. AB - Management of an indeterminate pulmonary nodule is a diagnostic challenge that commonly confronts primary care physicians and specialists. Patients often present with this radiographic finding in the course of an unrelated medical evaluation. We examined our institution's experience with percutaneous biopsy of lung nodules to determine the impact of this procedure on overall patient care. Although significant complications are uncommon, the expedience of percutaneous lung biopsy often supplants a surgical opinion prior to initiation of therapy without added diagnostic benefit or cost-savings. Hence, we caution practitioners to use this technique as an adjunct to diagnosis and not a substitute for multidisciplinary care. PMID- 15347107 TI - Transureteral saline perfusion to obtain renal hypothermia: potential application in laparoscopic partial nephrectomy. AB - BACKGROUND: Partial nephrectomy for resection of renal tumors often requires renal artery clamping and external renal cooling using ice-slush. Laparoscopic surgery precludes traditional ice-slush cooling. To facilitate renal cooling during laparoscopic partial nephrectomy, we investigated a method of intrarenal cooling by retrograde transureteral iced saline perfusion. METHODS: Open laparotomy was performed in 6 pigs. After atraumatic renal artery clamping, one kidney was cooled externally by using standard ice-slush; the other was cooled transureterally. For transureteral cooling, the ureter was cannulated with a double lumen 12 Fr catheter. Chilled saline (4 degrees C) irrigation was flushed through the catheter into the renal pelvis (16.7 mL/min) and allowed to drain via the second lumen of the catheter. Using a 30-gauge hypodermic thermometer, kidney temperatures were measured at 5-minute intervals for 30 minutes at 3 locations and 2 depths (0.5 cm and 1.5 cm). The animals were euthanized, and the kidneys were harvested for histologic examination. RESULTS: Renal cooling was achieved with both external and transureteral cooling. However, lower (5.0 versus 26.1 degrees C, P<0.001) parenchymal temperatures were achieved more rapidly with external renal cooling. During transureteral cooling, medullary (1.5 cm) temperatures were lower than cortical (0.5 cm) temperatures were; this difference did not reach statistical significance. CONCLUSIONS: Although renal hypothermia can be achieved by transureteral iced saline infusion, external cooling by using ice-slush appears to be more efficient in the porcine model. With refinement of the technique, intrarenal cooling via a transureteral approach may allow more effective cooling of the renal medulla, and limit warm ischemia during laparoscopic partial nephrectomy. PMID- 15347108 TI - Laparoscopic pyeloplasty and flexible nephroscopy: simultaneous treatment of ureteropelvic junction obstruction and nephrolithiasis. AB - BACKGROUND AND OBJECTIVE: Ureteropelvic junction obstruction and concomitant calculus disease may coexist. Therapeutic controversy exists regarding their ideal management. We report our use of flexible nephroscopy during laparoscopic pyeloplasty for caliceal stone removal. METHODS: From August 1998 through May 2002, 50 laparoscopic pyeloplasties were performed. Seven patients had documented ureteropelvic junction obstruction and ipsilateral nephrolithiasis. Preoperative stone burden and location were assessed. After pyelotomy, a 16 Fr flexible endoscope was passed through the uppermost trocar under direct laparoscopic guidance into the collecting system. Stone extraction was performed with a 2.4 Fr Nitinol basket. Postoperative imaging was assessed. RESULTS: Complete stone-free status confirmed by postoperative imaging was achieved in 6 of 7 patients. The longest individual stone diameter ranged from 4 mm to 13 mm (mean, 10.3 mm), and an average of 2.5 stones per patient was removed (range, 1 to 4 stones). Neither intraoperative fluoroscopy nor lithotripsy was required. No intraoperative or delayed complications were noted during a mean follow-up of 8.5 months (range, 2 to 17 months). CONCLUSIONS: Laparoscopic pyeloplasty and concomitant flexible nephroscopy with basket extraction is a simple, attractive alternative for the simultaneous treatment of ureteropelvic junction obstruction presenting with coexisting nephrolithiasis. It appears more efficacious when the stone number is limited and diameters measure from 5 mm to 20 mm. PMID- 15347109 TI - Microlaparoscopic-assisted vaginal hysterectomy in the morbidly obese patient. AB - OBJECTIVE: The purpose of this study was to demonstrate a minimally invasive, novel variation, microlaparoscopic-assisted vaginal hysterectomy (MAVH) of a previously established technique, laparoscopic-assisted vaginal hysterectomy (LAVH), in the morbidly obese patient. METHODS: This was a prospective, descriptive feasibility study (Canadian Task Force classification 11-2) conducted at a university-affiliated hospital and private community hospital. Seven morbidly obese women with an average body mass index of 45.8 kg/m2 (range, 40.6 to 51.5) underwent microlaparoscopic-assisted vaginal hysterectomy (MAVH). Microlaparoscopic-assisted vaginal hysterectomy (MAVH), classified as Type 1B, including unilateral or bilateral occlusion and division of the ovarian artery(ies), either medial or lateral to the ovary(ies), with or without dissection of the adjacent broad ligament under microlaparoscopic guidance, in addition to incision of the vesicouterine peritoneum. RESULTS: The median duration of surgery was 109.1 minutes (range, 86 to 134), median blood loss was 207 mL (range, 100 to 350), and average length of stay in the hospital was 33.7 hours (range, 23 to 48). The complication rate was 0%. CONCLUSION: Microlaparoscopic-assisted vaginal hysterectomy (MAVH) is a safe and effective, more minimally invasive method of performing laparoscopic hysterectomies in select morbidly obese patients. PMID- 15347110 TI - Laparoscopic management of bleeding after laparoscopic or vaginal hysterectomy. AB - OBJECTIVE: To assess the results and contributions of laparoscopy in the management of postoperative bleeding following laparoscopic (LH) or vaginal hysterectomy (VH). METHODS: A retrospective study of a 5-year period was carried out on 1167 women who underwent laparoscopic or vaginal hysterectomy. Ten women with postoperative bleeding following laparoscopic or vaginal hysterectomy were identified. RESULTS: The overall incidence of bleeding after laparoscopic or vaginal hysterectomy was 0.85% (10 of 1167). Over the 5-year study period, the incidence fluctuated between 1.1% and 0.4%. Surgical revision was primarily vaginal in 1 woman, followed by laparoscopic control. In 6 patients, laparoscopy was performed immediately. The patients profited from the prompt laparoscopic treatment, because intraabdominal hemorrhage was found and stopped. Of 6 cases of intraperitoneal bleeding, 1 resulted from a blood disorder. The collagen-fibrin agent TachoComb was applied locally, and the patient was postoperatively treated with blood products and coagulation factors. Only bipolar coagulation, TachoComb, and Foley catheter were used to achieve local hemostasis during laparoscopy. The remaining 3 cases where the vaginal cuff was bleeding were managed by vaginal repair and packing without laparoscopy. CONCLUSION: The laparoscopic approach to postoperative bleeding following laparoscopic or vaginal hysterectomy is an attractive alternative to the abdominal surgical approach. Bleeding following laparoscopic or vaginal hysterectomy can be managed by laparoscopy in the majority of patients. Because the abdominal incision is avoided, the recovery time is reduced. PMID- 15347111 TI - Robotically assisted laparoscopic Roux-en-Y hepaticojejunostomy. AB - INTRODUCTION: This study evaluates the feasibility and safety of using robotically assisted laparoscopy to perform a Roux-en-Y hepaticojejunostomy. This new method was compared with the open and standard laparoscopic approaches. METHODS: Eighteen pigs underwent a needlescopic common bile duct ligation to create a jaundice model. Three to 5 days later, transabdominal ultrasound was performed, and the common bile duct diameter was documented. For the Roux-en-Y hepaticojejunostomy, the pigs were randomly assigned to the open group (n=6), standard laparoscopy group (n=6), or robotically assisted laparoscopy group (Zeus) (n=6). One surgeon performed all 3 approaches with 1 assistant. Operative times, techniques, and complication rates were documented. RESULTS: The open approach was faster in all instances. At the hepaticojejunostomy, no difference was noted between the groups with the total number of stitches used. The robot required fewer stitches and less time in the posterior wall of the hepaticojejunostomy (P=-0.0083 and P=0.02049, respectively). The hepaticojejunostomy time was similar for the laparoscopy and robotically assisted groups. CONCLUSION: Robotically assisted laparoscopic Roux-en-Y hepaticojejunostomy is a feasible procedure. When compared with standard laparoscopy, operating time is similar. PMID- 15347112 TI - A technique for laparoscopic-assisted colectomy using two ports. AB - OBJECTIVES: Laparoscopic colon and rectal surgery requires advanced laparoscopic skills. The aim of this study was to describe a novel technique for laparoscopic assisted colectomy using only 2 ports and to review our initial experience with this technique for patients with benign colonic pathologies. METHODS: A retrospective chart review of all patients who had laparoscopic-assisted colon surgery using this technique was performed. The technique is described. RESULTS: For right colectomy, a 10-mm trocar for the camera was placed just below the umbilicus and a 5-mm working port just above the umbilicus. The colon was mobilized using one instrument and gravity assistance. The incisions were then connected, and the mobilized colon was pulled through this incision. For left sided colectomy, the 5-mm working port was placed at the left suprapubic hairline, which was then extended for removal of the specimen. Sixty patients with benign colonic pathologies had laparoscopic-assisted colon surgery using only 2 ports. Conversion to open surgery was required in 4 cases. The average length of the skin incision was 3.82 cm, and the mean length of hospital stay was 4.18 days. Postoperative complications occurred in 11 patients (18%) and included anastomotic leak in 1 patient and wound infection in 2. CONCLUSIONS: Laparoscopic assisted segmental colectomy using 2 ports is easy and feasible, with minimal skin incisions and fast recovery. Our initial experience suggests that it may be easier for the experienced colorectal surgeon to acquire the skills needed to perform this technique. PMID- 15347113 TI - Outcome and patient acceptance of outpatient laparoscopic cholecystectomy. AB - OBJECTIVE: The aim of this prospective study was to evaluate patients' experience and the outcome of outpatient laparoscopic cholecystectomy performed by a single upper gastrointestinal surgeon at a district hospital. METHODS: Between November 1999 and May 2003, 100 patients underwent outpatient laparoscopic cholecystectomy. Patients were followed up at 2 weeks as outpatients, and a questionnaire was mailed to all patients to assess their experiences. RESULTS: None of the patients required conversion to open cholecystectomy. One patient required admission to the hospital following drain insertion, and one patient was readmitted for pain control. One patient developed an epigastric port infection that resolved with antibiotics. Sixty-eight of the 100 patients completed the postal questionnaire. Thirty-five patients rated their overall experience as excellent. Twenty-three patients experienced very mild or no pain. All patients' right upper quadrant pain subsided or improved following surgery except one patient who stated that it became worse. Sixty-three patients (92.7%) stated they would recommend outpatient laparoscopic cholecystectomy to a friend or relative. CONCLUSION: Laparoscopic cholecystectomy can be performed safely as an outpatient procedure with a high acceptance and satisfaction rate in select patients. PMID- 15347114 TI - Evaluation of fundus-first laparoscopic cholecystectomy. AB - OBJECTIVES: Laparoscopic cholecystectomy is the gold standard for gallbladder surgery. Cholecystectomy from the fundus to the cystic duct may be advantageous when cystic duct exposure becomes difficult due to adhesions on Calot's triangle. The aim of this study was to compare conventional laparoscopic cholecystectomy with the fundus-first procedure and to evaluate whether the fundus-first technique can prevent conversion in difficult cases. METHODS: The study included 145 patients treated over 18 months. The inclusion criterion was the presence of ultrasound proven gallstones. Patients were excluded from the study if there was evidence of common bile duct stones, a bilioenteric fistula, or carcinoma of the gallbladder. RESULTS: The fundus-first approach was started in 45 patients; all procedures were completed laparoscopically. Conventional laparoscopic cholecystectomy was begun in 100 patients. Twenty-seven of the 100 patients were converted to fundus dissection (adhesions within Calot's triangle). Four of the 27 were further converted to open surgery. One patient had a drop in blood pressure on creation of pneumoperitoneum. Time taken for severely inflammatory and noninflammatory cases was significantly greater (P<0.05) in the fundus-first group. The average hospital stay was 48 hours in both groups. No major complications were observed. CONCLUSION: The rate of conversion in the conventional laparoscopic cholecystectomy group decreased from 18.75% (27/144) to 2.08% (3/144). The fundus-first technique has the potential to decrease conversion in difficult cases. PMID- 15347115 TI - Application of Doppler technology as an aid in identifying vascular structures during laparoscopy. AB - BACKGROUND: Intraoperative ultrasound has been used extensively during open surgery to assess bowel viability, to identify vascular structures, and to assess for congenital abnormalities. The extension of this technology in laparoscopic procedures has been hampered by the size of the equipment and the significant learning curve that accompanies its use. METHODS: Using a readily available Parks Inst. Co. Doppler Probe (8.1 MHz) and a 15-inch section of thick-walled, 9.5-mm OD Stainless Steel tubing, a Laparoscopic Doppler Probe was constructed. The parts were separately gas-sterilized, and a small segment of Penrose drain was used to create an airtight seal. The probe was passed through a 10-mm port, allowing assessment of vascular structures. RESULTS: Two Laparoscopic Doppler Probes were available for evaluation during a 1-month period at our hospital. Surgeons were then surveyed at the end of the 1-month period as to the utility of the devices. CONCLUSIONS: The Laparoscopic Doppler probe was used to identify the cystic artery during gallbladder dissection, to assess mesenteric blood vessels during laparoscopic colectomy, and to identify femoral vessels during laparoscopic preperitoneal hernia repair. It was found to be quick to construct, easy to use, and provided useful information to the operating surgeon. PMID- 15347116 TI - Prevention of de novo adhesion by ferric hyaluronate gel after laparoscopic surgery in an animal model. AB - BACKGROUND AND OBJECTIVE: Adhesions remain a major cause of severe long-term complications. Attempts have been made to prevent adhesion formation by using endogenous or exogenous materials with controversial results. Our aim was to evaluate the efficacy of 0.5% ferric hyaluronate gel in the prevention of adhesion formation after laparoscopic surgery. METHODS: This was a prospective, randomized, experimental study (animal model). The study population comprised 75 female rabbits (Fauve de Bourgogne) weighing over 3 kg. The rabbits were randomized into 3 groups of 25 (hyaluronate, saline, and control) by using a predetermined computer-generated randomization code. All rabbits underwent a peritoneal laparoscopic resection, and the main outcome measure was the adhesion formation after laparoscopic surgery. RESULTS: The laparoscopic operating time and the mean interval before second-look surgery were not different among the 3 groups. The number of rabbits with adhesions did not differ among the 3 groups. The bowel adhesion rate was higher at the 10-mm trocar site than at the 5-mm trocar site (P=0.01). The adhesion scores did not differ among the 3 groups. A strong correlation was found between the values of the different adhesion scoring systems used. CONCLUSION: These results obtained in a rabbit model suggest that routine intraperitoneal application of hyaluronate gel does not prevent adhesion formation after laparoscopic surgery. PMID- 15347117 TI - Laparoscopic repair of a spigelian hernia: a case report and literature review. AB - BACKGROUND: We present a case in which laparoscopy was both diagnostic and therapeutic in a patient with a spigelian hernia. CASE REPORT: A 35-year-old man was referred to the General Surgery Service for evaluation of right lower quadrant abdominal pain of approximately 6 months. The pain was not disabling but was a constant discomfort. The patient did not have any significant past medical or surgical history, and the physical examination was significant only for an area of focal tenderness in the right lower quadrant. Ultrasound and CT scans of the patient's abdomen were unremarkable. A laparoscopic exploration of the area revealed a defect in the area of semilunar and semicircular lines consistent with a spigelian hernia. The patient underwent a laparoscopic herniorrhaphy with placement of a polypropylene mesh. CONCLUSION: This case illustrates the role of laparoscopy in the diagnosis and treatment of spigelian hernias. PMID- 15347118 TI - Splenic injury following endoscopic retrograde cholangiopancreatography: a case report and review of the literature. AB - Endoscopy of both the upper and lower gastrointestinal tract remains one of the commonest procedures performed on patients. The common risk factors associated with this procedure are well known. However, some of the rarer complications are not known or are not considered in the immediate postprocedure time period when a patient experiences acute abdominal pain. Herein, we report a case of solid organ injury (splenic capsular tear) resulting from endoscopic retrograde cholangiopancreatography. The risk factors and presentation of this type of injury are reviewed, including its association with chronic pancreatitis, which we describe for the first time. PMID- 15347119 TI - Laparoscopic reduction of appendicocecal intussusception due to mucinous cystadenoma in an adult. AB - OBJECTIVE: Laparoscopic reduction of appendicocecal intussusception due to mucinous cystadenoma is herein described. METHODS: A 32-year-old female had intermittent abdominal pain and nonbloody diarrhea. The preoperative diagnosis was intussusception of the colon with a benign tumor by CT-scan findings. RESULTS: Laparoscopic reduction of intussusception and ileocecal resection were successfully performed. Gauze sponge sticks were used to provide compression from the distal colon. This maneuver was very effective to reduce the intussusception. CONCLUSION: Laparoscopic reduction of intussusception due to a benign tumor is feasible and safe even in adults. Mucinous cystadenoma is one of the causes of intussusception in adults and a good indication for the laparoscopic approach. PMID- 15347120 TI - Clipless cholecystectomy: broadening the role of the harmonic scalpel. AB - BACKGROUND: The ultrasonically activated (Harmonic) scalpel has proven to be an effective, efficient, and safe instrument for dissection and hemostasis in both open and laparoscopic surgical procedures. To date, the primary use of the Harmonic scalpel in laparoscopic cholecystectomies has been for the division of the cystic artery and liver bed dissection. Advancements in the Harmonic scalpel blade tip now provide for the reliable ultrasonic division and closure of the cystic duct. METHODS: In a personal, prospective series involving 100 consecutive patients undergoing laparoscopic cholecystectomies, the Harmonic scalpel was used as the sole instrument for division of the cystic duct and artery as well as dissection of the liver bed. Two patients with large cystic ducts (over 5 mm) received an additional ductal ligature. RESULTS: No patients developed postoperative hemorrhage or bile leakage. CONCLUSION: The Harmonic scalpel provides complete hemobiliary stasis for most patients and is a safe alternative to standard clip or ligature closure of the cystic duct. Furthermore, there may be a cost savings inherent in a procedure utilizing a single disposable instrument. PMID- 15347121 TI - Laparoscopic-assisted resection of a large retroperitoneal tumor. AB - BACKGROUND: Retroperitoneal tumors have traditionally been excised using standard open techniques. When preparing the manuscript we knew of only 1 report, in Japanese, describing the excision of 2 retroperitoneal neurofibromas using a minimally invasive technique. The feasibility of the technique was tested in the removal of a large retroperitoneal neurofibroma. METHODS: Laparoscopic-assisted resection of a large retroperitoneal tumor was offered to a 37-year-old female with a 12 x 10 x 9-cm neurofibroma located inferior to the left kidney. The patient was placed on a beanbag in a right lateral decubitus position. Three 5-mm ports were used, and the tumor was mobilized followed by placement of a hand port for completion of dissection and removal of the tumor. RESULTS: The tumor was successfully removed in about 3 hours, and blood loss was <100 cc. The patient developed urinary and bowel dysfunction probably secondary to the effect of the dissection and removal of the tumor on the nearby sacral plexus. CONCLUSIONS: We believe that laparoscopy is a feasible and appropriate technique available to general and laparoscopic surgeons in the management of retroperitoneal tumors, providing excellent visualization and adequate access for proper dissection through small incisions. PMID- 15347122 TI - Retroperitoneoscopic lumbar sympathectomy for buerger's disease: a novel technique. AB - BACKGROUND: The conventional lumbar sympathectomy procedure through the extraperitoneal route requires a muscle cutting-splitting incision, which leads to significant postoperative pain and prolonged convalescence. With increasing experience in retroperitoneoscopic procedures, we did a pilot study to explore the role of retroperitoneoscopy in lumbar sympathectomy. We describe herein our technique used for the surgery. METHODS: The patient was placed in a lateral position. A 15-mm incision was made just below the 12th rib, and retroperitoneal space was created using blunt finger dissection. A custom-made, large balloon was inserted and inflated with the equivalent of 750 mL to 1000 mL of saline. The second 10-mm port was placed in line with the first port above the iliac crest. The third and fourth 5-mm ports were placed anterior to the first 2 ports. Peritoneum was retracted anteriorly. The medial border of the psoas muscle was used as a landmark and a chain identified immediately medial to it. Lumbar vessels were ligated on the right side. The first to fourth lumbar sympathetic ganglia were removed with the intervening chain. The port sites were closed without a drain. RESULTS: We attempted and successfully completed this procedure in 8 patients; 6 on the left side and 2 on the right side. The average operating time was 38 minutes. The mean hospital stay was 1.5 days. All patients had symptomatic pain relief and clinical improvement. CONCLUSIONS: Retroperitoneoscopic lumbar sympathectomy is a safe and effective procedure. It has a short convalescent time and minimal morbidity; hence, it is a viable alternative for the open procedure. PMID- 15347123 TI - Single and biplane TrueFISP cardiovascular magnetic resonance for rapid evaluation of left ventricular volumes and ejection fraction. AB - INTRODUCTION: Cardiovascular magnetic resonance (CMR) allows very accurate, but time-consuming, volume assessment by the short-axis slice summation technique. The single and biplane methods of volume assessment are used less, partly because FLASH cine imaging provides poor blood-myocardium contrast in long-axis views. TrueFISP gives excellent blood-myocardium contrast, even in patients with heart failure. We hypothesized that the single plane and biplane methods of volume assessment in TrueFISP images might provide an acceptable degree of accuracy and be quicker than the short axis method, and that single and biplane left ventricular volume assessment would be more accurate with TrueFISP than with FLASH in patients with impaired ventricular function. METHODS: Short- and long axis CMR images were obtained by FLASH and TrueFISP with a 1.5-T scanner. We determined the accuracy of both single and biplane long-axis methods for left ventricular volume and ejection fraction (EF) measurements compared with the conventional short-axis method in 10 heart failure patients using both FLASH and TrueFISP and in 9 healthy subjects using TrueFISP. RESULTS: No difference in volumes and EF was found between the single plane method, the biplane method, and the short-axis method using TrueFISP for image acquisition, in both patients and healthy subjects. The same was true of the results obtained by FLASH in the patients with heart failure. CONCLUSIONS: The single and biplane methods, regardless of whether TrueFISP or FLASH is used, are a reasonable and rapid alternative to the conventional short-axis approach for left ventricular volume and EF assessment in patients with heart failure and impaired ventricular function. PMID- 15347124 TI - Right ventricular wall motion abnormalities found in healthy subjects by cardiovascular magnetic resonance imaging and characterized with a new segmental model. AB - AIM: To evaluate right ventricular wall motion abnormalities in healthy subjects using a new segmental model for the right ventricle. METHODS AND RESULTS: 29 healthy subjects (9 female, 20 male, mean age 48.9+/-15 years) underwent cardiovascular magnetic resonance imaging (CMR; 1.5-Tesla Sonata, Siemens, Erlangen, Germany) for the evaluation of cardiac function and right ventricular wall motion. A steady-state free precession gradient-echo sequence (TrueFISP) was used. Right ventricular wall motion was analyzed, and the site of areas of disordered motion was classified according to the new segmental model. Such areas were seen in 27 (93.1%) of the 29 subjects. Dyskinesia was found in 22 subjects (75.9%), hypokinesia in 11 (37.9%), and bulging in 8 (27.6%). The number of wall motion abnormalities diagnosed was significantly higher in the transverse plane (86.2%) than in the short-axis plane (13.8%) and the horizontal longitudinal plane (41.4%; p = 0.000). CONCLUSION: Right ventricular wall motion abnormalities are one of the criteria for the diagnosis of arrhythmogenic right ventricular cardiomyopathy. However, our findings indicate that they may also be seen around the insertion of the moderator band in healthy subjects, so that the significance of their presence at this site in patients undergoing diagnostic investigations for this disease should be interpreted with caution. PMID- 15347125 TI - Evaluation of a new method for automated detection of left ventricular boundaries in time series of magnetic resonance images using an Active Appearance Motion Model. AB - The purpose of this study was the evaluation of a computer algorithm for the automated detection of endocardial and epicardial boundaries of the left ventricle in time series of short-axis magnetic resonance images based on an Active Appearance Motion Model (AAMM). In 20 short-axis MR examinations, manual contours were defined in multiple temporal frames (from end-diastole to end systole) in multiple slices from base to apex. Using a leave-one-out procedure, the image data and contours were used to build 20 different AAMMs giving a statistical description of the ventricular shape, gray value appearance, and cardiac motion patterns in the training set. Automated contour detection was performed by iteratively deforming the AAMM within statistically allowed limits until an optimal match was found between the deformed AAMM and the underlying image data of the left-out subject. Global ventricular function results derived from automatically detected contours were compared with results obtained from manually traced boundaries. The AAMM contour detection method was successful in 17 of 20 studies. The three failures were excluded from further statistical analysis. Automated contour detection resulted in small, but statistically nonsignificant, underestimations of ventricular volumes and mass: differences for end-diastolic volume were 0.3%+/-12.0%, for end-systolic volume 2.0%+/-23.4% and for left ventricular myocardial mass 0.73%+/-14.9% (mean+/-SD). An excellent agreement was observed in the ejection fraction: difference of 0.1%+/-6.7%. In conclusion, the presented fully automated contour detection method provides assessment of quantitative global function that is comparable to manual analysis. PMID- 15347126 TI - The role of still-frame parametric imaging in magnetic resonance assessment of left ventricular wall motion by non-cardiologists. AB - BACKGROUND: Cardiac magnetic resonance (MR) images are often reviewed by non cardiologists who are not trained in the interpretation of regional left ventricular (LV) function. We hypothesized that the use of still-frame parametric MR images of wall motion could aid in the assessment of regional LV function. METHODS: Dynamic, electrocardiogram-gated, steady-state free precession (FIESTA) short-axis images were obtained in 6 to 10 slices in 18 consecutive patients. Each loop was used to automatically generate a still-frame image, in which each pixel is assigned a value equal to the amplitude of cyclic variation in local intensity, resulting in higher intensity in pixels that change between blood and tissue during the cardiac cycle. The dynamic images were reviewed by an expert cardiologist who provided gold standard grades for regional wall motion and by four radiologists. Then the radiologists reviewed and graded the same MR images in combination with parametric images. Grades assigned to each segment in the two sessions were compared with the gold standard. RESULTS: According to expert interpretation, 6 patients had normal wall motion, and 12 had wall motion abnormalities. Parametric images showed a bright band in the area spanned by endocardial motion, with reduced brightness and thickness in areas of hypokinesis. The agreement between the radiologists' grades and the gold standard significantly improved by adding parametric images (from 77% to 81%), which also resulted in reduced interobserver variability (from 52% to 33%). CONCLUSIONS: Still-frame parametric images aid in the assessment of regional wall motion by non-cardiologists who are required to interpret cardiac images. PMID- 15347127 TI - Three-directional myocardial motion assessed using 3D phase contrast MRI. AB - Regional myocardial function is a complex entity consisting of motion in three dimensions (3D). Besides magnetic resonance imaging (MRI), no other noninvasive technique can give a true 3D description of cardiac motion. Using a time-resolved 3D phase contrast technique, three-dimensional image volumes containing myocardial velocity data in six normal volunteers were acquired. Coordinates and velocity information were extracted from nine points placed in different myocardial segments in the left ventricle (LV), and decomposed into longitudinal (V(L)), radial (V(R)), and circumferential (V(C)) velocity components. Our findings confirm a longitudinal apex-to-base gradient for the LV, with only a small motion of the apex. The mean velocity for V(L) for all the basal segments was higher compared to the midsegments during systole [3.5+/-1.2 vs. 2.5+/-1.7 cm/s (p<0.01)], early filling [-6.9+/-1.8 vs. -4.9+/-1.8 cm/s (p<0.001)], and during atrial contraction [-2.2+/-1.4 vs. -1.6+/-1.3 cm/s (p<0.05)]. A similar pattern was observed when comparing velocities from the midsegments to the apex. Radial velocity was higher during early filling in the midportion of the lateral [-4.9+/-2.7 vs. -3.2+/-1.6 cm/s (p<0.05)] wall compared to the basal segments, no difference was observed for the septal [-2.0+/-1.5 vs. -0.3+/-2.5 cm/s (p=0.15)], anterior [-5.8+/-3.3 vs. -4.0+/-1.7 cm/s (p=0.17)], and posterior [-2.3+/-2.1 vs. -2.5+/-1.0 cm/s (p=0.78)] walls. When observing the myocardial velocity in a single point and visualizing the movement of the main direction of the velocities in this point as vectors in velocity vector plots like planes, it is clear that myocardial movement is by no means one dimensional. In conclusion, our time resolved 3D, phase contrast MRI technique makes it feasible to extract myocardial velocities from anywhere in the myocardium, including all three velocity components without the need for positioning any slices at the time of acquisition. PMID- 15347128 TI - Measurement of aortic blood flow by magnetic resonance below and above the origin of the coronary arteries in postmenopausal hormone replacement therapy. AB - PURPOSE: Principal blood flow measures might be assessable by velocity-encoded cine magnetic resonance (VENC MR) of aortic blood flow. The feasibility of using VENC MR for clinical research was tested in a contemporary and controversial human model: the effects of 17beta-estradiol (E) and cyclic norethisterone acetate (NETA) in postmenopausal women. METHODS: Sixteen postmenopausal women were included in a crossover study (two 12-week periods, 3-month washout) and randomized to E and NETA or placebo. Aortic blood flow (ABF) was measured eight times by VENC MR below and above the coronary arteries (CA) and was used for estimation of coronary artery blood flow (CABF) and peak flow velocity (Vmax). The calculated value of CABF was negative and was corrected by averaging systolic flow. Data were analyzed in a repeated measurement model including analysis of repeatability (CR) and group variability (CV). RESULTS: The CR and CV of ABF were low (11% and 7%) and corresponded at the two levels. Vmax showed similar levels of reproducibility at the two levels. The coronary artery blood flow was less reproducible (39% and 31%). During treatment, ABF above CA was reduced after 12 weeks (p=0.03), ABF below CA was unchanged, and CABF was significantly increased without detrimental effects of NETA. Vmax was increased at NETA addition (p<0.01). CONCLUSIONS: Aortic flow quantification by VENC MR is reproducible and useful for assessment of principal haemodynamic changes in smaller studies. Derived measurement of CABF lacks precision. Differences in change below and above CA indicate that oral high-dose E and NETA may induce coronary artery dilatation. PMID- 15347129 TI - Cardiac involvement in Mulibrey nanism: characterization with magnetic resonance imaging. AB - Mulibrey nanism (MUL) is an autosomal recessive disorder that is enriched in the Finnish population. Variable degrees of pericardial and myocardial involvement can lead to heart failure and premature death. The purpose of this study was using magnetic resonance imaging (MRI) to assess structural and functional abnormalities of the MUL cardiopathy in all four cardiac chambers as well as in the pericardium. Thirty-one patients with MUL (mean age 27, range 15-50 years) and 16 controls (mean age 31, range 19-45 years) were examined with a Siemens Vision 1.5-T imager. Ten patients had undergone pericardiectomies to relieve symptoms of constrictive pericarditis. In surgery performed 0.5-25 years before MRI, the removed pericardium was found to be thickened and consisting of scarlike fibrosis. Turbo spin echo images were obtained for assessment of pericardial thickness, and breath hold left ventricular (LV) short axis and four-chamber cine images were obtained for the volumetric data. In MRI, pericardial thickness was normal (under 3.4 mm) in all patients with MUL. In the 10 pericardiectomized patients, the remnants of the pericardium were of normal thickness as well. The LV septum (p = 0.01) and posterior wall (p<0.001) were hypertrophied and end diastolic volumes of both ventricles (p<0.05) were reduced in all patients. The LV systolic function was preserved. The volume chance during the first third of diastole (p = 0.030), the absolute peak filling rate (p = 0.047), and the time to peak rate of LV diastolic filling (p = 0.030) indicated restrictive LV diastolic filling. The right ventricular ejection fraction and contraction of both atria were reduced. PMID- 15347130 TI - Anteroseptal or apical myocardial infarction: a controversy addressed using delayed enhancement cardiovascular magnetic resonance imaging. AB - AIM: Delayed enhancement MRI (DE-MRI) of the heart has been shown to reliably identify areas of irreversible myocardial damage. We sought to determine if the term anteroseptal MI is appropriate by correlating electrocardiographic, angiographic, cine MRI and DE-MRI findings. METHODS AND RESULTS: Nineteen patients admitted to our hospital with their first acute anterior MI and whose ECG showed new Q waves in leads V1-V4 were studied. All patients underwent cardiac catheterization, cine MRI, and DE-MRI. The mean left ventricular ejection fraction was 53%+/-16%. All 19 patients had evidence of delayed hyperenhancement in one or more myocardial segments (mean number of affected segments 5.5+/-2.1). The mean mass of hyperenhanced myocardium was 14+/-8 grams, or 10%+/-6% of absolute LV mass. Nineteen (100%) and 15 (79%) patients showed evidence of delayed hyperenhancement of the apex and apical anterior segments respectively. Seven (37%) patients showed evidence of mid ventricular anteroseptal hyperenhancement and none had any hyperenhancement of basal anteroseptal segments. CONCLUSION: High resolution cardiac MRI applied in patients with acute infarction and new Q waves in leads V1-V4 demonstrates the presence of predominantly apical, but not isolated septal or anteroseptal infarction. PMID- 15347131 TI - Three-dimensional, time-resolved motion of the coronary arteries. AB - BACKGROUND: Coronary artery motion can decrease image quality during coronary magnetic resonance angiography and computed tomography coronary angiography. PURPOSE: To characterize the three-dimensional motion of the coronary arteries along the entire vessel length and to identify the temporal location and duration of periods of relatively low cardiac motion in patients with coronary artery disease. METHODS: Archived digital, biplane x-ray angiography films acquired at 30 frames per second with simultaneous electrocardiogram recording were reviewed for 15 patients with coronary artery disease. The right coronary (RCA), left anterior descending (LAD), and left circumflex (LCX) arteries were divided into proximal, mid, and distal segments. The displacement and velocity of a point in each segment were calculated throughout the heart cycle. Time-dependent, three dimensional motion of each segment on each vessel was determined. Periods of the heart cycle during which maximal displacement was less than 1 mm or 0.5 mm per frame for each artery were determined. RESULTS: A period lasting an average of 187 msec was seen during mid-diastole (72+/-5% of the cardiac cycle) in which all three coronary arteries studied had relatively little motion. This period of quiescence was consistent along the length of the arteries. Although the amount of motion did vary along the length of the arteries, there was no difference in the timing of rest periods in the proximal, mid, and distal segments using a < 1 mm per frame threshold. The periods of low motion were significantly reduced in length and often altogether eliminated when the 0.5 mm per frame threshold was used. PMID- 15347132 TI - Quantification of atherosclerosis with MRI and image processing in spontaneously hyperlipidemic rabbits. AB - The need for a quantitative method to assess atherosclerosis in vivo is well known. This study tested, in a familiar animal model of atherosclerosis, a combination of magnetic resonance imaging (MRI) and image processing. Six spontaneously hyperlipidemic (Watanabe) rabbits were examined with a knee coil in a 1.5-T clinical MRI scanner. Inflow angio (2DI) and proton density weighted (PDW) images were acquired to examine 10 cm of the aorta immediately cranial to the aortic bifurcation. Examination of the thoracic aorta was added in four animals. To identify the inner and outer boundary of the arterial wall, a dynamic contour algorithm (Gradient Vector Flow snakes) was applied to the 2DI and PDW images, respectively, after which the vessel wall area was calculated. The results were compared with histopathological measurements of intima and intima media cross-sectional area. The correlation coefficient between wall area measurements with MRI snakes and intima-media area was 0.879 when computed individual-wise for abdominal aortas, 0.958 for thoracic aortas, and 0.834 when computed segment-wise. When the algorithm was applied to the PDW images only, somewhat lower correlations were obtained. The MRI yielded significantly higher values than histopathology, which excludes the adventitia. Magnetic resonance imaging, in combination with dynamic contours, may be a suitable technique for quantitative assessment of atherosclerosis in vivo. Using two sequences for the measurement seems to be superior to using a single sequence. PMID- 15347133 TI - Detection of metabolic abnormality in asynergic regions of ischemic human myocardium using 31P and 1H magnetic resonance spectroscopy. AB - To assess quantitatively phosphocreatine (PCr), adenosine triphosphate (ATP) and total creatine (CR) in asynergic regions of ischemic human myocardium using phosphorus (31P) and proton magnetic resonance spectroscopy (1H MRS). Patients were divided into two groups: 31P MRS and 1H MRS. In each group, patients were subdivided into three groups using echocardiography: a normokinetic [WN(P) (n = 6) in 31P MRS, WN(H) (n = 10) in 1H MRS], a hypokinetic [WH(P) (n = 13), WH(H) (n = 7)], and a- or dyskinetic wall motion groups [WA(P) (n = 14), WA(H) (n =6)]. They were compared with normal subjects of each group [CNP (n = 10), CN(H) (n = 10)]. 31P MRS spectra were obtained from the anterior and apical regions of the left ventricle by slice-selected 1D CSI. 1H MRS spectra were obtained from the 2 x 2 x 2-cm voxel set in the left ventricular wall by the PRESS method. In the 31P MRS group, myocardial PCr was significantly lower in the WH(P) and WA(P) groups than in the CN(P) group, but myocardial PCr in the WN(P) group did not differ from that in CN(P). A difference in ATP could not be detected among the four groups. In the 1H MRS group, myocardial CR was significantly lower in the WH(H) and WA(H) groups than in the CN(H) group. Myocardial CR in the WNH group did not differ from that in the CN(H). The noninvasive 31P and 1H MRS approach is useful in the assessment of metabolite reduction associated with wall motion abnormality. PMID- 15347134 TI - Thallium-201 SPECT and low-dose dobutamine stress cine MRI for predicting functional recovery of regional myocardial contraction in patients with myocardial infarction. AB - OBJECTIVES: The purpose of this study was to compare the diagnostic performances of Tl-201 single photon emission computed tomography (SPECT) and dobutamine stress cine magnetic resonance imaging (MRI) for predicting functional recovery of regional myocardial contraction in patients after myocardial infarction. METHODS: Twenty patients underwent Tl-201 SPECT and MRI 3-4 weeks after onset of myocardial infarction. Cine MR images were acquired in the resting state and during dobutamine stress. Tl-201 uptake and systolic wall thickening (SWT) on cine MRI were analyzed on short-axis images by using a 14-segment model. Follow up cine MR images were obtained 187.1+/-33.5 days after onset. RESULTS: The averaged Tl-201 uptake in 54 segments with impaired SWT was 47%+/-20%, being significantly lower than that in 226 segments with preserved SWT (75%+/-18%; p<0.0001). The sensitivity, specificity, and accuracy of dobutamine MRI and Tl 201 SPECT for predicting preserved SWT after 6 months were 89% vs. 80%, not significant (NS); 89% vs. 72%, p<0.01; and 89% vs. 79%, NS, respectively. In the anterior wall and apex, the sensitivity and specificity of SPECT were not significantly different from those of MRI. In the inferior wall and posterolateral wall, however, the specificity of SPECT was substantially lower than that of MRI (53% vs. 88%, p<0.001), resulting in significantly lower accuracy (75% vs. 90%, p<0.01). CONCLUSIONS: Both SPECT and dobutamine MRI showed excellent sensitivity for predicting myocardial viability in all left ventricular segments. Decreased specificity of SPECT in the inferior and posterolateral segments resulted in lower overall specificity in comparison with dobutamine MRI. PMID- 15347135 TI - Off-frequency tuning error artifact in steady-state free precession cine imaging due to adjacent air-filled bowel. AB - Identification of common cardiovascular magnetic resonance (CMR) artifacts is important to optimal image acquisition quality and the avoidance of mistaken clinical diagnosis. We describe an off-frequency tuning error artifact observed during rapid steady-state free precession acquisitions in three patients and resulting in a dramatic appearance within the right and left ventricles. These artifacts were associated with large, adjacent air pockets within a loop of bowel or stomach and were eradicated by retuning of the magnetic resonance scanner's frequency. Awareness of this artifact, its cause and correction, should improve diagnostic image quality and avoid clinical diagnostic confusion. This report also heightens the need for a more robust shimming sequence for cardiac studies. PMID- 15347136 TI - Fmoc-based chemical synthesis and selective binding to supercoiled DNA of the p53 C-terminal segment and its phosphorylated and acetylated derivatives. AB - The C-terminal domain of p53 comprises a linker, the tetramerization domain and the regulatory domain, and contains at least seven sites of potential post translational modification. An improved strategy was developed for the synthesis of large peptides that contain phosphorylated amino acids and p53(303-393), a 91 amino acid peptide, and three post-translationally modified derivatives were synthesized through the sequential condensation of three partially protected segments. Peptide thiolesters were prepared using the sulfonamide-based 'safety catch' resin approach and employing Fmoc-based solid-phase peptide synthesis. At the N-terminus of the middle building block, a photolabile protecting group, 3,4 dimethoxy-6-nitrobenzyloxycarbonyl, was incorporated to differentiate the N terminal amino group from the side-chain amino groups. Two sequential couplings were accomplished following this protection strategy. The synthetic products, p53(303-393) and its phosphorylated or acetylated derivatives, exhibited the ability to bind specifically to supercoiled DNA, which is one of the characteristics of this domain. PMID- 15347137 TI - The relative orientation of the Arg and Asp side chains defined by a pseudodihedral angle as a key criterion for evaluating the structure-activity relationship of RGD peptides. AB - The ability of an integrin to distinguish between the RGD-containing extracellular matrix proteins is thought to be due partially to the variety of RGD conformations. Three criteria have been proposed for the evaluation of the structure-activity relationship of RGD-containing peptides. These include: (i) the distance between the charged centres, (ii) the distance between the Arg Cbeta and Asp Cbeta atoms, and (iii) the pseudo-dihedral angle defining the Arg and Asp side-chain orientation formed by the Arg Czeta, Arg Calpha, Asp Calpha and Asp Cgamma atoms. A comparative conformation-activity study was performed between linear RGD peptides and strongly constrained cyclic (S,S) -CDC- bearing compounds, which cover a wide range of inhibition potency of platelet aggregation. It is concluded that the fulfilment of the -45 degrees < or = pseudo dihedral angle < or = +45 degrees criterion is a prerequisite for an RGD compound to exhibit inhibitory activity. Once this criterion is accomplished, the longer the distance between the charged centres and/or between the Arg and Asp Cbeta atoms, the higher is the biological activity. In addition, the stronger the ionic interaction between Arg and Asp charged side chains, the lower the anti aggregatory activity. PMID- 15347138 TI - Hybrid alpha/beta3-peptides with proteinogenic side chains. Monosubstituted analogues of the chemotactic tripeptide For-Met-Leu-Phe-OMe. AB - The alpha/beta3-mixed tripeptides R-CO-beta3-HMet-Leu-Phe-OMe (1a,b), R-CO-Met beta3-HLeu-Phe-OMe (2a,b) and R-CO-Met-Leu-beta3-HPhe-OMe (3a,b) (a, R = tert butyloxy-; b, R = H-), analogues of the potent chemoattractant For-Met-Leu-Phe OMe, have been synthesized by classical solution methods and fully characterized. The activities of the new analogues as chemoattractants, superoxide anion producers and lysozyme releasers have been determined on human neutrophils. Whereas all of the three N-formyl derivatives are significantly less active than the parent tripeptide as chemoattractants, compound 1b has been found to be highly active as a superoxide anion producer and 3b as a lysozyme releaser. The results show that the replacement of the native Leu residue at the central position is, in each of the examined cases, the least favourable modification. The three N-Boc derivatives are, as expected, devoid of activity as agonists, but they are all good inhibitors of chemotaxis. Information on the solution conformation has been obtained by examining the involvement of the NH groups in intramolecular H-bonds using 1H NMR. The conformation of the N-Boc analogue 1a has also been determined in the crystal state by x-ray diffraction analysis. The molecule is extended at the beta3-HMet residue (phi1 = -87 degrees; theta1 = 172 degrees; psi1 = 126 degrees) and no intramolecular H-bond is present. PMID- 15347139 TI - Potent trypsin-resistant hGH-RH analogues. AB - A series of analogues of hGH-RH-(1-29)-NH2 designed to have metabolic stability has been synthesized. Standard Boc-SPPS was employed, modified to permit the guanidinylation of amino side-chains after chain assembly but before release from the resin. [Dat1, Har(11, 12, 20, 21, 29), Ala15, Nle27, Asp28]-, [Dat1, Har(11, 20, 29), Orn12, Ala15, Nle27, Asp28]-, and [Dat1, Gap(11,12, 21, 29), Ala15, Har20, Nle27, Asp28]-hGH-RH-(1-29)-NH2 were completely resistant to trypsin and about 50 times as potent as hGH-RH-(1-29)-NH2 itself when injected subcutaneously in rats. These peptides are candidates for clinical application in the therapy of GH deficiency. PMID- 15347140 TI - Xenotransplantation--state of the art. PMID- 15347141 TI - Xenotransplantation: biotechnological aspects and current attitudes. AB - Xenotransplantation of organs from the large domestic species will only be successful if the donor animals have been genetically modified, in particular regarding the alpha-Gal epitope, certain human complements (CD55 and CD59) and/or H-transferase. This requires, among other things, major embryo-technological efforts, and the rate of success is still far from an acceptable level in the domestic species. It is currently poor, but the progress is very good. In this brief review certain embryo-technological problems will be addressed with the focus on the pig as potential organ donor. In addition, certain views of the Danish ad hoc Committee on Gene Technology on xenotransplantation will be presented in this context as they are supposed to mirror the concern and the views of the issues important for the public and each individual. PMID- 15347142 TI - Xenotransplantation--clinical activities and regulatory development. PMID- 15347143 TI - The use of fish in xenotransplantation research. PMID- 15347144 TI - Possible transmission of zoonoses in xenotransplantation: porcine endogenous retroviruses (PERVs) from an immunological point of view. PMID- 15347145 TI - Xenotransplantation and a potential threat of animal pathogens in humans. PMID- 15347146 TI - Embryonic and adult stem cells. PMID- 15347147 TI - Rearing and caring for a future xenograft donor pig. PMID- 15347148 TI - Custom made piggeries in Norway? View of the meat producing industry. PMID- 15347149 TI - Political considerations of controversial medical issues: xenotransplantation and society. A presentation of the work of the Norwegian national working group on xenotransplantation. PMID- 15347150 TI - Xenotransplantation--view of the transplanted patient. PMID- 15347152 TI - Xenotransplantation--the donor welfare perspective. PMID- 15347151 TI - Xenotransplantation--ethical considerations based on human and societal perspectives. PMID- 15347153 TI - Going for brokers: Spitzer rides again. PMID- 15347154 TI - Hill Physicians Medical Group outshines California's pay-for-performance program. PMID- 15347155 TI - The weight debate. PMID- 15347156 TI - Managed Medicaid business might be worth the difficulties. PMID- 15347157 TI - 10 things you want from your PBM. PMID- 15347158 TI - NCQA's new online survey tries to tame the paper tiger. PMID- 15347159 TI - Matthew Holt: Internet pundit thrives on the biggest issues. PMID- 15347160 TI - Cost-effective management of hyperglycemia in patients with type 2 diabetes using oral agents. AB - Diabetes exacts an enormous toll on health care resources, with extremely high costs attributable to care of diabetes patients in proportion to the afflicted population. Though individual treatment strategies are required for each patient, newer long-acting sulfonylureas may be the initial drugs of choice, as they may be the only oral agents that inhibit the processes inducing hyperglycemia- hepatic glucose production and glucose utilization by the tissues--by improving insulin secretion and insulin resistance. Sulfonylureas also represent the most cost-effective therapeutic option, alone or in combination with other oral agents or insulin. The newer long-acting agents, glimepiride and glipizide GITS, may be more attractive among sulfonylureas, due to their greater insulin-sparing property, fewer hypoglycemic events, weight neutrality, and once-daily dosing. Glimepiride may be preferred due to its safety profile, especially for the elderly and those with hepatic and/or renal dysfunction. PMID- 15347161 TI - Nesiritide speeds recovery time for heart failure patients. PMID- 15347162 TI - Plans wait for payoff from IT investment. PMID- 15347163 TI - [Quality assurance of the diagnosis of osteoporosis]. PMID- 15347164 TI - [Breakthrough in the treatment of osteoporosis]. PMID- 15347165 TI - [Reducing blood loss in total knee arthroplasty]. PMID- 15347166 TI - [Teriparatide (human recombinant 1-34 parathyroid hormone)--a new treatment regimen for severe osteoporosis]. PMID- 15347167 TI - [Quality assurance of the diagnosis of osteoporosis in a county]. PMID- 15347168 TI - [Leptospirosis in Denmark, 1980-2002. A relevant diagnosis for Danish physicians?]. PMID- 15347169 TI - [Public outpatient alcohol treatment]. PMID- 15347170 TI - [Familial adenomatous polyposis. Results from the Polyposis Registry through 25 years]. PMID- 15347172 TI - [Palliative pain relief]. PMID- 15347171 TI - [A randomized comparison of carvedilol and metoprolol in patients with chronic heart failure. The Carvedilol Or Metoprolol European Trial (COMET) ]. PMID- 15347173 TI - [Rabies. A case of supposed exposure]. PMID- 15347174 TI - [Subacute onset non-hereditary lymphedema praecox]. PMID- 15347175 TI - [Picture of the month: Meckel diverticulum]. PMID- 15347176 TI - [Does TV watching cause child behavior disorders?]. PMID- 15347177 TI - [Arthroscopy, deep infection and survey of results 2]. PMID- 15347178 TI - [Treatment of primary biliary cirrhosis with ursodeoxycholic acid without liver biopsy?]. PMID- 15347179 TI - [By whom are adults with hyperkinetic disorders/ADHD/"DAMP" to be treated?]. PMID- 15347180 TI - [Use of acupuncture in primary health care]. PMID- 15347181 TI - [Allergy tests]. PMID- 15347182 TI - [Laparoscopic or open inguinal hernia surgery?]. PMID- 15347183 TI - [The thesis about the powerless placebo]. PMID- 15347184 TI - [They promise one thing and promise to keep something else]. PMID- 15347185 TI - [The new internship education]. PMID- 15347186 TI - ["The gout physician" should give up unreasonable solutions]. PMID- 15347187 TI - [Primary liver disease in the horse]. AB - The clinical signs of liver disease are highly variable and non-specific. Irrespective of the cause or the duration of liver disease, more specific clinical signs, e.g. hepatic encephalopathy, become apparent in the advanced stages of the disease. Due to the non-specific clinical signs, the possible diagnosis of liver disease is frequently not taken into consideration. However, measurement of the plasma or serum concentrations of total bile acids and gamma glutamyl transferase (gamma GT) may provide valuable diagnostic information. The specific diagnosis can be confirmed by ultrasound examination of the liver and histological examination of a liver biopsy specimen. The most frequently documented liver diseases are acute hepatic necrosis, chronic hepatitis caused by pyrrolizidine alkaloids, cholelithiasis, and haemochromatosis. Immediate treatment with antibiotics and polyionic fluids, in association with supportive nutritional care, is usually necessary to maintain the horse until sufficient liver regeneration occurs to provide adequate function. The use of corticosteroids is contraindicated, except for patients with chronic active hepatitis. In some cases, e.g. portosystemic shunts or cholelithiasis, surgical intervention is indicated. Liver disease may be more common than is currently appreciated since little accurate information on the prevalence or incidence is available. PMID- 15347188 TI - [For God's sake]. PMID- 15347189 TI - [Infectious bovine keratoconjunctivitis (IBK) in cows, clinical and lab review at four farms]. AB - After several reports to the GD (Dutch Animal Health Service) from practitioners in The Netherlands concerning serious Infectious Bovine Keratoconjunctivitis (IBK) in dairy herds during summer and autumn 2003, the GD has carried out a pilot-study to determine the most responsible agent. This pilot was thought to be important because of the painfulness of the illness and problems like, (for the farmer) an intensive and difficult therapy. Also the report of a Chlamydophilae infection causing IBK in a dairy herd in the UK prompted to this study. The most frequently isolated infectious agent in our study was Moraxella, probably M. bovis. For the presence of Chlamydophila, mycoplasmata or BHV1 viruses were no indications. PMID- 15347190 TI - [Veterinarian Marlice Vonck after 3 months of the KNMvD: "A lot is happening in there"]. PMID- 15347191 TI - [Advisory council will give pedigreed pups an identification chip at an earlier age]. PMID- 15347192 TI - [FIDIN antibiotics report: a comment]. PMID- 15347193 TI - [A reaction to the discussion about chemotherapy]. PMID- 15347194 TI - Distribution and speciation of metals in surface sediments of Taranto [corrected] gulf (Ionian Sea, Southern Italy). AB - Concentrations of metals (Al, Fe, Ti, Ni, Mn, Cr, Zn, Cu, Pb) were determined in surface sediments from Taranto gulf (Ionian Sea, Southern Italy) in order to assess the distribution and the bioavailability of metals. Total metal content and speciation of Fe, Ni, Mn, Zn, Cu and Pb were determined. Sediment cores were also sampled in some stations of particular interest. The analytical results show that metals distribution does not show significant variations in relation to depth. Metal distribution in the Taranto gulf is mainly influenced by industrial wastes from Taranto town, by rivers of the Basilicata region and by prevailing anticlockwise marine currents. These factors cause an accumulation of metals in the sediments near to the coasts of Calabria. The statistical analysis, effected by HCA and PCA methods, can be considered a good starting point in order to classify sediment samples and to locate accumulation areas. Metal speciation shows that Fe, Ni and Zn are present in sediments mainly as sulphides, Mn is present principally in an ion-exchange form or bound to carbonate, while Cu and Pb are mainly present in the fraction bounds to humic acids. PMID- 15347195 TI - Heavy metals in marine coastal sediments: assessing sources, fluxes, history and trends. AB - Examples are presented from the Adriatic Sea, the Ligurian Sea and the Venice Lagoon to illustrate different approaches to the study of anthropogenic metals in marine coastal sediments. These examples refer to studies of areal distribution and transport mechanisms, individuation of the sources, sediment dating, chronology of the fluxes, present and past trends. In particular, some of the findings achieved in studying the Venice Lagoon are discussed from the point of view of anthropogenic changes both in sediment composition and contaminant fluxes. PMID- 15347196 TI - The role of organic matter on metal toxicity and bio-availability. AB - A short review concerning the role of organic matter on metal toxicity and bio availability in aqueous systems is carried out. The complexity of the issue derives both from the high number of natural and anthropogenic organic compounds and from the variability of their structures. In fact, the binding capacity and affinity is dependent on the number and type of ligands, on their position in the structures, on the ligand/metal ratio. It is also necessary to develop analytical protocol in order to carry out speciation studies of organic carbon and of metals bound to organic compounds, and at the same time to characterise the nature of the complexes. PMID- 15347197 TI - Importance and difficulties of research on metal speciation in the aquatic ecosystem: an ecologist's viewpoint. AB - Post-war research on the role of heavy metals in the environment was stimulated by a new sensitivity towards nature, the increasing impact of human activities on the environment, continuous progress in analytical chemistry and the development of radioecology. Nowadays, very low concentrations of total metal can be quantified in the various compartments of the ecosystem (e.g. water, sediment, suspended particles, plants and animals), and the physical and chemical forms of the metal identified. A current difficulty is how to evaluate the role of the various metal forms in the natural ecosystem. The major issues to be tackled are: variations over time of metal species in relation to the influence of environmental variables; the fact that the availability of the metal varies with the plant and animal species; the ability of the organism to select the metals and their forms; the influence of suspended particles and chelating substances on metal uptake, detoxification strategies and synergic and antagonistic effects. The advantages and disadvantages of monitoring using bioaccumulator organisms are discussed. PMID- 15347198 TI - Bioavailability, bioaccumulation and tolerance of chromium: consequences in the food chain of freshwater ecosystems. AB - Many abiotic and biotic factors can influence the partitioning equilibrium of heavy metals, thus influencing metal impact on aquatic environments. Unicellular algal species release soluble organic substances able to complex metals. In our laboratory a Cr-tolerant strain was selected and isolated from a wild type strain of Scenedesmus acutus. The exudates released by the two strains counteracted the growth inhibition caused by Cr(VI) and the exudates of the Cr-tolerant strain were more effective. On the contrary, the exudates did not reduce chromium toxicity to the cladoceran Daphnia magna. The reduction of chromium effect on algae seems the consequence of an algae-specific interaction among Cr(VI), exudates and algal cells. Chromium uptake resulted to be energy-dependent since bioaccumulation rate in subdued light condition was lower than at high light intensity. The effect of Cr(VI) on algae changed depending on metabolism of the cells and in particular it seemed to be related to the bioaccumulation rate. Tolerance in the selected strain could not be ascribed to a lower uptake of chromium. The difference in sensitivity to chromium between the two strains was exploited to evaluate if tolerance acquired by algae could have consequences for Daphnia. After treatment with Cr(VI), the two strains of S. acutus were used as food source for D. magna. The results indicate that chromium is accumulated by algae in a form not available for daphnids and that Cr tolerance acquired by the algae can be of some advantage to the consumer organism. PMID- 15347199 TI - The use of diffusive gradients in thin-film (DGT) passive samplers for the measurement of bioavailable metals in river water. PMID- 15347201 TI - Study of the distribution of trace elements in six species of marine organisms of the Ligurian Sea (north-western Mediterranean)--comparison with previous findings. AB - This paper presents the results of the determination of several trace elements (total and organic mercury, selenium, cadmium, lead, copper, manganese and zinc) in six species of marine organisms (Nephrops norvegicus, Engraulis encrasicolus, Mullus barbatus, Mullus surmuletus, Merluccius merluccius, and Sarda sarda), sampled in the Ligurian Sea (North-West Mediterranean) during a three-year period, from 1998 to 2000. The species analysed (388 samples) are the same studied in previous investigations in the same area (260 samples), so that a comparison with previous data measured in 1977-1986 was possible, in order to evaluate the variation of the marine environment in that area with respect to that period. Biological parameters of the specimens (length and weight of the individuals, and also sex for N. norvegicus) were taken into account in order to make possible the comparison between old and new data. No appreciable variation occurred in the levels of the trace elements analysed during the period 1998 2000, while the comparison with the data collected about 25 years ago showed a decrease of all the elements except for selenium. PMID- 15347200 TI - FT-IR spectroscopy and chemometrics as a useful approach for determining chemical physical properties of gasoline, by minimizing analytical times and sample handling. AB - Since recently Mid-Infrared Spectroscopy is being used in the petrol industry for determining important chemical-physical properties of gasoline. This technique enables to achieve results similar to those obtainable with standard methods at shorter times, lower costs and safer sample handling. This can be obtained by employing multivariate regression models in the framework of a chemometric analytical approach. In the present work some gasoline properties including density, Reid vapor pressure, Research Octane Number, aromatic hydrocarbons content and fraction recovered at 70 degrees C were determined with standard methods (ISO, CEN, ASTM) on 93 samples of unleaded gasoline produced in an oil refinery. The same samples were also analyzed by FT-IR spectrophotometry, recording absorptions of seven characteristic bands. The obtained values of chemical-physical and spectroscopic properties were used to develop and validate a multivariate regression model. This established an experimental correlation between these two sets of variables derived from the same gasoline samples. The model was able to correctly predict the chemical physical properties of a set of unknown gasoline samples produced in the same refinery starting from the spectroscopic ones. PMID- 15347202 TI - Adsorption of organic compounds onto activated carbons from recycled vegetables biomass. AB - The removal of organic species from aqueous solution by activated carbons is investigated. The latter ones are prepared from olive husks and almond shells. A wide range of surface area values are obtained varying temperature and duration of both carbonization and activation steps. The adsorption isotherm of phenol, catechol and 2,6-dichlorophenol involving the activated carbons prepared are obtained at 25 degrees C. The corresponding behavior is quantitatively correlated using classical isotherm, whose parameters are estimated by fitting the equilibrium data. A two component isotherm (phenol/2,6-dichlorophenol) is determined in order to test activated carbon behavior during competitive adsorption. PMID- 15347203 TI - Comparing some operational approaches to the limit of detection. AB - The limit of detection is one of the fundamental metrological characteristics of any chemical measurement systems but, even now, it is a highly controversial matter because of the existing differences in terminology, formulations and understanding. Even when limiting the analysis to those operational approaches that, because of their simplicity, are often suggested by the guidelines of some international bodies, the available approaches can return appreciably different estimates. In this paper, the matter is critically reconsidered, and the different approaches compared with the aid of some case studies. PMID- 15347204 TI - Spectrophotometric studies on the complexation equilibria of Ni and Pb with 2 carboxy-2-hydroxy-5'-sulfo-formazylbenzene. Simultaneous determination of trace amounts of Ni and Pb. AB - Spectrophotometric studies have been made to investigate the reaction of Nickel and Lead with 2-carboxy-2'-hydroxy-5'-sulfoformazyl-benzene (zincon) in 50%(v/v) ethanol-water at 25 degrees C and an ionic strength of 0.1 M NaClO4. A complete picture of the complexation equilibria in the pH range (4.2-12.0) for nickel and (1.9-11.5) for lead are presented. Simple, rapid, selective and sensitive methods for the spectro-photometric determination of nickel and lead has been developed based on the color reaction of their complexes with zincon. The methods allow the determination of 4.69 microg mL(-1) of nickel at pH = 6.3 (lambdamax = 665 nm) and 10.3 microg ml(-1) of lead at pH = 5.6 (lambdamax = 610 nm). The apparent molar absorptivities were epsilon = 1.3 x 10(4) L mol(-1) cm(-1) for nickel and epsilon = 0.6 x 10(4) L mol(-1) cm(-1) for lead. The interference of a large number of foreign ions and complexing agents has been studied. Thiosulphate, as masking agent allows the simultaneous determination of nickel and lead in the presence of high concentrations of copper. Ascorbic acid, sodium cyanide and or sodium fluoride provide the elimination of many other interferences. The methods have been applied successfully to the simultaneous determination of nickel and lead in an aluminium and non-ferrous alloy. PMID- 15347205 TI - Synergistic extraction of cobalt (II) by beta hydroxy naphthaldoxime and neutral donors. AB - Liquid - liquid extraction of Cobalt (II) from aqueous hydrochloric acid medium by the use of oxime derivative of beta-hydroxy naphthaldehyde in o-xylene medium is being reported. The ligand was synthesized and characterized in our laboratory. The typical pH range of extraction of Co (II) by the ligand - donor combination was seen to be between 8 - 9. The effect of different donors like dimethyl sulphoxide (DMSO), trioctyl phosphine oxide (TOPO) and bis(2-ethylhexyl) phosphonate and the effect of various diluents on the extraction of Co(II) by the present method were studied in detail. The binary adduct formation constant (log kex) in the organic phase was found to be 3.182. The overall equilibrium constant (log K) for the ternary species [Co(A)(DMSO)(Cl)], [Co(A)(TOPO)(Cl)] and [Co(A)(phosphonate)(Cl)] were estimated to be 6.42, 6.22 and 6.25 respectively. The trend in equilibrium constants were in accordance with their basic character, i.e., (DMSO > or = TOPO approximately/= phosphonate). The results were extrapolated in the determination of Cobalt in pharmaceutical drugs and cobalt bearing ore samples. PMID- 15347206 TI - Zero-crossing second derivative spectrophotometry for simultaneous determination of palladium and nickel using 1-(2-pyridylazo)-2-naphthol in mixed micellar solutions. AB - 1-(2-Pyridylazo)-2-naphthol (PAN) has been used for the simultaneous determination of nickel and palladium at trace levels. PAN complexes of nickel and palladium in the pH 1.98 form red and green colored complexes, respectively, which are soluble in aqueous 4:1 Triton X-100 to sodium dodecylsulfate (SDS) micellar media with total detergent concentration of 3.2%. Under optimum conditions, calibration graphs for the simultaneous determination by second derivative spectrophotometry were obtained. Zero crossing second derivative spectrophotometry at 668 and 572 nm, respectively for palladium and nickel was used for the simultaneous determination. The method is able to determine palladium to nickel ratio 70:1 to 1:6 (Wt/Wt), accurately. Accuracy and reproducibility of the determination method on the known various amounts of palladium and nickel in their binary mixtures were tested. Effects of diverse ions on the determination of palladium and nickel to investigate selectivity of the method also were studied. PMID- 15347207 TI - Inhibition of acidic corrosion of carbon steel by some mono and bis azo dyes based on 1,5 dihydroxynaphihalene. AB - Inhibition of the corrosion of carbon steel in hydrochloric acid solution by some mono- and bis-azo dyes based on 1,5-dihydroxynaphthalene was studied in relation to the concentration of inhibitors using weight loss and potentiostatic polarization techniques. The percentage inhibition efficiency calculated from two methods is in a good agreement with each other. The inhibition mechanism of the additives was ascribed to the formation of complex compound adsorbed on the metal surface. The adsorption process follows Freundlich adsorption isotherm. The formation of the complex compound was studied by conductometric and potentiometric titrations. The stability constants of the Fe-complexes were determined using the latter technique and related to the inhibition efficiency. PMID- 15347208 TI - Monitoring of volatile hydrocarbon compounds (BTEX) in the town of Viterbo and indoor-outdoor evaluation in domestic sites. PMID- 15347209 TI - Use of a large angioplasty balloon for predilation is a risk factor for embolic complications in protected carotid stenting. AB - Periprocedural neurological complications (PNCs) after carotid stenting were retrospectively analyzed to determine the risk factors with the use of various protective devices. Forty-three lesions in 40 patients were treated by carotid stenting with distal balloon protection for nearly all postdilation procedures and some predilation procedures. The following variables were statistically analyzed for association with PNCs: diameter of the angioplasty balloon used for predilation, use of a distal protection balloon during predilation, use of a protection balloon during postdilation, lesion-bifurcation distance, length of the lesion, age, clinical presentation of the lesion (symptomatic or asymptomatic), and hypercholesterolemia. PNCs occurred in five patients, four with minor deficits and one with major deficits. Univariate analysis showed large diameter of the predilation angioplasty balloon (p = 0.0026), use of a protection balloon during predilation (p = 0.0075), lesion length (p = 0.0003), and lesion bifurcation distance (p = 0.0006) were significantly associated with PNCs. Multivariate analysis of these four variables showed that use of a large angioplasty balloon for predilation was the only independent predictor (p = 0.004, odds ratio 34.00) for the occurrence of PNCs. Use of a large angioplasty balloon for predilation carries the risk of periprocedural embolic complications. Therefore, even when a protection device is used, predilation should be performed with a small balloon. PMID- 15347210 TI - High titer retroviral gene transduction to neural progenitor cells for establishment of donor cells for neural transplantation to parkinsonian model rats. AB - Neural progenitor cells (NPCs) are expected to be useful donor sources for cell transplantation therapy in Parkinson's disease. However, control of the differentiational lineage, especially into dopaminergic neurons, is still difficult. Thus, genetic modification of NPCs to produce l-dopa is potentially useful. The present study prepared high titer retrovirus carrying human tyrosine hydroxylase-1 (HTH-1) gene. HTH-1 gene could be efficiently transduced into NPCs obtained from the E12.5 rat mesencephalon. This retroviral gene transduction caused no apparent changes in survival, proliferation, or differentiation. In vitro, HTH-1 gene-transduced NPCs released little l-dopa and addition of tetrahydrobiopterin, the cofactor of tyrosine hydroxylase, was required for production of l-dopa. In vivo, three of seven hemi-parkinsonian model rats that received HTH-1 gene-transduced donor NPCs achieved functional recovery. High titer retroviral vector for gene transduction could be used to prepare NPCs for transplantation to hemi-parkinsonian model rats. However, functional recovery after transplantation of HTH-1 gene-transduced NPCs was incomplete. PMID- 15347211 TI - Manual occipital ventricular puncture for cerebrospinal fluid shunt surgery: can aiming be standardized? AB - The manual occipital ventricular puncture is a standard surgical procedure, but specific targeting has not received sufficient attention despite the experience of anatomical disorientation. This study tried to identify an exact site for the ideal trajectory with this method, especially in the sagittal plane, which avoids contact with the choroid plexus that may be the major source of complications. A total of 44 consecutive adult cases undergoing cerebrospinal fluid shunting through the occipital route for hydrocephalus were retrospectively reviewed for the following: correlations between burr hole site, direction of puncture, and location of the ventricular catheter based on postsurgical radiological studies; calculation of the ideal trajectory to place the catheter tip in the anterior horn of the lateral ventricle without contact with the choroid plexus through the standard occipital burr hole. In addition, the relationships between the location of the ventricular structure, the cranial base line connecting the nasion and inion, and the size of the ventricle were evaluated. Incorrect catheter emplacement was found in five cases, which suggested that this procedure using the standard burr hole site and aim point might result in incorrect catheter placement. The ideal external aim points were widely distributed because of the variable heights of ventricular structures from the cranial baseline. No correlation between the locations of the anatomical points and ventricular size was found. The present study could not define a standard external aim point. Tailored preoperative planning of the trajectory is recommended. PMID- 15347212 TI - Shaken baby syndrome manifesting as chronic subdural hematoma: importance of single photon emission computed tomography for treatment indications--case report. AB - A boy with shaken baby syndrome first presented at age 3 months with acute subdural hematoma (SDH) and was treated by subdural tapping at a local hospital. Chronic SDH was identified at a rehabilitation center at age 19 months. The chronic SDH appeared to have developed within the preceding 16 months. His physical and mental development was already delayed. Magnetic resonance (MR) imaging revealed a 20-mm thick right chronic SDH with midline shift and small bilateral subdural effusions. The chronic SDH had compressed the right cerebral hemisphere. MR imaging also disclosed bilateral cerebral atrophy. 99mTc ethylcysteinate dimer single photon emission computed tomography (SPECT) revealed decreased cerebral blood flow (CBF) in the non-hematoma hemisphere, although CBF is said to decrease on the chronic SDH side, especially if midline shift is present. Burr hole craniotomy with external drainage was performed, but the patient showed no change in CBF postoperatively, although the volume of hematoma decreased. The patient was clinically unchanged immediately after the operation. In this case, SPECT measurement of CBF was important in evaluating the pathophysiology of the delays in physical and mental growth. Atrophy of the bilateral hemispheres was the major mechanism in the decreased CBF, not the compression by chronic SDH. MR imaging and SPECT can determine the surgical indications for chronic SDH in patients with cortical atrophy. PMID- 15347213 TI - Ruptured vertebral artery-posterior inferior cerebellar artery aneurysm associated with pulseless disease--case report. AB - A 48-year-old woman with a 29-year history of pulseless disease presented with subarachnoid hemorrhage caused by a rare ruptured intracranial aneurysm of the right vertebral-posterior inferior cerebellar artery. The aneurysm was successfully clipped. Pulseless disease is a chronic and nonspecific vasculitis involving mainly the aorta and its main branches such as the carotid, subclavian, vertebral, and coronary arteries. The cervical vessels are mostly affected in Japanese patients. Almost all patients have ischemic disorders of the involved vessels, manifesting as syncope, visual disturbance, or a faint or absent pulse. The mechanisms responsible for pulseless disease and intracranial aneurysms are likely to be different. PMID- 15347214 TI - Middle cerebral artery aneurysm associated with Behcet's disease--case report. AB - A 37-year-old man with Behcet's disease suffered secondary subarachnoid hemorrhage and intracerebral hematoma in the right temporal lobe caused by a ruptured aneurysm of the right middle cerebral artery. The aneurysm was successfully clipped. Aneurysm formation is common in the visceral arteries in Behcet's disease, but extremely rare in the intracranial arteries. Vasculitis may be involved in the etiology of intracranial aneurysms in patients with Behcet's disease. PMID- 15347215 TI - Middle cerebral artery thrombolysis through the contralateral internal carotid artery--case report. AB - A 63-year-old male presented with sudden onset of right hemiplegia and global aphasia. On admission he was stuporous. Computed tomography (CT) revealed no abnormalities except for right intraventricular meningioma found incidentally. Emergency angiography confirmed complete occlusion of the left internal carotid artery (ICA) and left M1 trunk whereas the left ICA bifurcation remained patent. The ipsilateral ICA was permanently occluded with two detachable balloons to prevent thrombus migration into the distal ICA and middle cerebral artery (MCA), followed by thrombolysis of the clot in the ipsilateral M1 through the contralateral ICA with urokinase (total dose 420,000 U) under systemic heparinization. Partial recanalization of the ipsilateral MCA was accomplished. The time interval from onset to recanalization was about 3 hours. Postoperative CT showed no hemorrhagic transformation. Slight right paresis and mild motor aphasia persisted 2 months later and he was transferred to a rehabilitation facility. Thrombolysis of the MCA embolism can be performed through the contralateral ICA in the presence of ipsilateral ICA occlusion. PMID- 15347216 TI - Diffusion-weighted magnetic resonance imaging of organized subdural hematoma- case report. AB - A 59-year-old male presented with a left organized subdural hematoma. The hematoma appeared as a homogeneous low density area on brain computed tomography and as hyperintense and isointense area on both fluid-attenuated inversion recovery and T2-weighted magnetic resonance (MR) imaging. Echo-planar diffusion weighted MR imaging showed a crescent hyperintense area under the dura mater and an irregular hypointense area over the brain surface in the left subdural space. The apparent diffusion coefficient (ADC) values of the solid and liquid hematoma were 0.86 +/- 0.32 x 10(-3) and 2.56 +/- 0.39 x 10(-3) mm2/sec, respectively. The ADC value of the solid hematoma was similar to acute subdural or intraparenchymal hematoma, and that of the liquid was similar to cerebrospinal fluid. Burr-hole surgery failed to remove all the hematoma, and he complained of persistent headache. The hematoma was removed through a craniotomy without further neurological deficits. Organized subdural hematoma often requires craniotomy for evacuation because of its solid content. Diffusion-weighted MR imaging and measurement of ADC values can differentiate solid from liquid hematoma, so are useful for selection of the surgical procedure. PMID- 15347217 TI - Ectopic growth hormone-releasing adenoma in the cavernous sinus--case report. AB - A 55-year-old woman presented a rare ectopic pituitary adenoma in the right cavernous sinus manifesting as acromegaly. The tumor was removed via transsphenoidal approach. Intraoperative observation showed the adenoma was located entirely within the right cavernous sinus, and separated from the normal pituitary gland by the medial wall of the cavernous sinus. There was no communication between the tumor and the pituitary. Histological examination showed a growth hormone-releasing adenoma. Including our case, only eight of 86 reported ectopic adenomas have occurred in the cavernous sinus. Such ectopic presentation may be responsible for failed transsphenoidal surgery for endocrinologically active tumors. PMID- 15347218 TI - Treatment of L5-S1 spondyloptosis with single-stage surgery through the posterior approach--case report. AB - A 21-year-old male patient with L5-S1 spondyloptosis was treated by total L-5 laminectomy with foraminotomy and posterior fusion through the posterior approach. His complaints of severe low back pain and limited spine mobility were resolved. No new deficits occurred. The surgical management of spondyloptosis includes one-, two-, or three-stage operations with posterior, anterior, or combined approaches. Careful posterior decompression and posterior fusion without reduction may be adequate for the treatment of L5-S1 spondyloptosis. PMID- 15347219 TI - Complications of revision spinal surgery. AB - Surgical treatment for symptomatic cervical and lumbar spondylosis has become prevalent in recent years. With this increased intervention, increasing numbers of patients experience persistent symptoms and require revision spinal surgery. Although many aspects of the workup and operation are similar for both primary and revision surgery, there are special considerations that must be examined when determining if a patient is a candidate for revision surgery. Preoperative workup should include evaluation for spinal instability. Intraoperatively, scar tissue may complicate the procedure, and care must be taken to avoid incidental durotomy. The prognosis for a revision surgery can be predicted best by the patient's outcome after the primary surgery. As with any surgical procedure, patient selection is imperative for successful outcome. PMID- 15347220 TI - The biomechanics of iatrogenic spinal destabilization and implant failure. AB - Revision spinal surgery is usually indicated in cases of persistent or recurrent symptoms related to neural compression, spinal deformity, or construct failure. An understanding of fundamental biomechanical principles of both spinal decompression and reconstructive strategies is essential to avoid unnecessary subsequent spinal operations. PMID- 15347221 TI - Revision surgery for Chiari malformation decompression. AB - Chiari malformations comprise four different hindbrain anomalies originally described by Hans Chiari, a professor of pathology at the German University in Prague. There are four basic Chiari malformations. The reasons for revision of Chiari malformation decompression may be for conservative or inadequate initial decompression or the development of postoperative complications. Another reason involves cases of both hindbrain herniation and syringomyelia in patients who have undergone adequate posterior fossa decompression without resolution of symptoms, signs, or radiological appearance of their syrinx cavity. Additionally, symptom recurrence has been reported in association with various types of dural grafts. Reoperation or revision surgery for patients with Chiari malformations is common and may not be due to technical error or inadequate decompression. The types of revision surgeries, their indications, and initial presentations will be reviewed. PMID- 15347222 TI - Management of strut graft failure in anterior cervical spine surgery. AB - Although successfully used, long strut grafts are vulnerable to dislodgment, displacement, fracture, and nonunion, which can require revision surgery; thus, meticulous preparation of the vertebral endplate along with exact sizing and harvesting of the bone graft with plating are essential for successful outcomes. Biomechanical data and previous clinical studies support the addition of posterior fusion and fixation following multilevel (more than two-level) corpectomy. The additional posterior instrumentation moves the instantaneous axis of rotation posteriorly, thus approximating its normal location in the posterior vertebral body (VB). Biomechanically, this protects the graft from excessive loads while in extension and explains the clinical success of circumferential instrumentation for long-segment corpectomy reconstructions. If strut fracture occurs with minimal displacement and the graft position is still satisfactory, application of a halo vest and judicious observation are recommended. Significant displacement, kyphosis, or loss of contact of the graft and VB require revision surgery. In patients requiring revision surgery for nonunion, placement of fibular autograft or allograft with use of bone morphogenetic protein is likely to be beneficial. If questions remain regarding bone quality or construct stability, the supplemental use of posterior stabilization is recommended. Various surgical approaches have been advocated for treatment of symptomatic anterior cervical pseudarthroses or nonunion. It remains controversial as to whether the anterior or posterior approach is best. Adequate understanding of the graft and implant biomechanics are essential for a successful outcome. PMID- 15347223 TI - Postlaminectomy cervical deformity. AB - Postlaminectomy cervical kyphosis is an important consideration when performing surgery. Identifying factors predisposing to postoperative deformity is essential. The goal is to prevent postlaminectomy cervical kyphosis while exposing the patient to minimal additional morbidity. When postlaminectomy kyphosis does occur, surgical correction is often required and performed via an anterior, posterior, or combined approach. The authors discuss the indications for surgical approaches as well as clinical results. PMID- 15347224 TI - Surgery for postarthrodesis adjacent-cervical segment degeneration. AB - Anterior cervical decompression and fusion has gained popularity because of its applicability to a variety of cervical spine disorders. The authors of long-term follow-up studies have demonstrated the development of degenerative changes in segments adjacent to fusion. So-called adjacent-segment disease causes symptomatic deterioration in up to 25% of the patients who have undergone anterior cervical decompression and fusion for cervical spondylotic myelopathy. The causes of this condition are debated in the literature. The authors provide a review of the available literature on the pathogenesis, prevention, and treatment of postarthrodesis adjacent-segment degenerative disease. PMID- 15347225 TI - Laminoplasty for the treatment of failed anterior cervical spine surgery. AB - OBJECT: Cervical stenotic myelopathy can be treated via anterior or posterior approaches. In anterior cervical decompression and fusion (ACDF), because the risks and likelihood of pseudarthrosis increase with the number of treated segments, attempts are typically made to limit the number of treated levels. Thus, postoperative recurrence of myelopathy following ACDF may occur because stenotic levels were not treated or because adjacent segments have degenerated. Revision decompressive surgery via an anterior approach is one solution; however, if the stenosis involves multiple levels a posterior decompressive laminoplasty can be performed as an alternative. METHODS: Twenty-four cases treated over an 8 year period were identified and data were retrospectively reviewed. In 15 cases posterior decompressive surgery was necessary because of progressive spinal degeneration and stenosis (five cases following initial treatment for radiculopathy, seven after initial treatment for spondylotic myelopathy, and three due to spreading of an ossified posterior longitudinal ligament). In nine cases revision surgery was undertaken because the initial decompression was inadequate. The mean follow-up period after the second surgery was 16 months. Improvements in myelopathy were seen in 83% of patients (mean improvement of 1.25 points on the Nurick Scale). Preoperative severe gait disorders were associated with poor recovery. Complications included two cases of transient C-5 nerve root palsy and two cases of new persistent axial neck pain. CONCLUSIONS: Laminoplasty is a straightforward and effective treatment for failed ACDF due to inadequate decompression or progressive degeneration of the spinal column, avoiding reentry through scar tissue. In terms of myelopathic pain, the recovery rate is comparable with that related to revision ACDF. PMID- 15347226 TI - Management of iatrogenic flat-back syndrome. AB - Iatrogenic loss of lordosis is now frequently recognized as a complication following placement of thoracolumbar instrumentation, especially with distraction instrumentation. Flat-back syndrome is characterized by forward inclination of the trunk, inability to stand upright, and back pain. Evaluation of the deformity should include a full-length lateral radiograph obtained with the patient's knees and hips fully extended. The most common cause of the deformity includes the use of distraction instrumentation in the lumbar spine and pseudarthrosis. Surgical treatment described in the literature includes opening (Smith-Petersen) osteotomy, polysegmental osteotomy, and closing wedge osteotomy. The authors will review the literature, cause, clinical presentation, prevention, and surgical management of flat-back syndrome. PMID- 15347227 TI - Revision strategies for lumbar pseudarthrosis. AB - Management and avoidance of lumbar pseudarthrosis are among the most common and challenging tasks faced by reconstructive spine surgeons. The risks of pseudarthrosis can be broadly divided into two categories: those within a surgeon's control and those not within his/her control. These include biological factors, graft choices, site preparation, and surgical design. The authors review the biological factors that affect fusion and how they can be manipulated to avoid or manage lumbar pseudarthrosis. Surgical planning and construct design to prevent or treat pseudarthrosis will also be discussed. Additionally, the importance of restoring sagittal balance will be reviewed. PMID- 15347228 TI - Recurrent lumbar disc herniation. AB - Recurrent lumbar disc herniation is a common disease process. It has been noted to occur in 5 to 15% of cases surgically treated for primary lumbar disc herniation. Outcomes in one series approached those after the initial operations, although this is not the case in the experience of most surgeons. The removal of recurrent lumbar disc herniations requires meticulous surgical technique. Great care is taken to identify the osseous margins of the previous surgical site. Identification and dissection of scar from the dura mater is greatly aided with the use of a microscope. PMID- 15347229 TI - Microendoscopic discectomy for recurrent disc herniations. AB - OBJECT: The use of microendoscopic discectomy (MED) for the treatment of primary lumbar disc herniations has become fairly well accepted; its role in recurrent disc herniations is less clear. The reluctance of many surgeons to use this technique stems, in part, from the concern of undertaking an endoscopic discectomy in a patient in whom the anatomy is distorted from a previous operation. It appears counterintuitive to operate through a limited working area when the traditional open approach for recurrence favors wider exposure of the surgical field. Given that operating on previously exposed tissue can be associated with even greater morbidity than on virginal tissue, the authors describe their experience with performing MED for recurrent disc herniation. METHODS: Unilateral MED was performed in patients with classic symptoms of lumbar radiculopathy, a previous operation at that level, and findings of recurrent disc herniation on magnetic resonance imaging. The approach was similar to a standard MED. Aided by fluoroscopic guidance, a working cannula was docked on the laminofacet junction at the level of the nerve root, with care taken to ensure a slightly more lateral initial trajectory. A good decompression of the nerve root could then be achieved through the use of the endoscope with preservation of the paraspinous musculature and much of the remaining facet capsule. Ten consecutive patients undergoing the procedure were analyzed prospectively and compared with the previous 25 who underwent routine single-level MED. Use of the MED technique provided excellent visualization and decompression of the nerve root; no conversions to open procedures were necessary in either group. The average operative time in the experimental group was 98.5 minutes, with a mean blood loss of 33 ml and an approximate hospital stay of 7.3 hours. In this respect, there was no statistical difference between the two groups (analysis of variance, p = 0.39, 0.68, and 0.51, respectively). There was one cerebrospinal fluid leak in each group. CONCLUSIONS: Microendoscopic discectomy for recurrent disc herniation can be safely performed without an increase in surgery related morbidity. PMID- 15347230 TI - Clinical outcomes after minimal-access surgery for recurrent lumbar disc herniation. AB - OBJECT: Experience with minimal-access surgical approaches for revision lumbar surgery has not been previously reported. METHODS: During a 7-month period, 10 consecutive patients with recurrent disc herniations underwent revision operations in which microendoscopic discectomy (MED) was performed. Perioperative data and clinical outcomes (according to Macnab criteria) were compared with those obtained in 25 consecutive patients who underwent routine single-level MED as well as with previously published data. Overall, outcome of the MED-treated revision group was excellent or good in 90% during a mean follow-up period of 18.5 months (minimum 12 months). Operative blood loss, duration, complications, and length of hospital stay were not significantly different between the revision and primary MED-treated groups. CONCLUSIONS: Analysis of these early data suggests equivalent or superior results are obtained when performing MED compared with historical controls in which conventional surgery was conducted for recurrent disc surgery. The procedure appears to be a safe and effective alternative in cases in which recurrent lumbar disc herniation causes radiculopathy. PMID- 15347231 TI - Lumbar myofascial flap for pseudomeningocele repair. AB - OBJECT: Initial management for lumbar pseudomeningoceles entails the closed external drainage of cerebrospinal fluid (CSF) with or without blood patch application. The presence of longstanding pseudomeningoceles and those associated with nonmicroscopic dural tears can be more problematic. Additionally the failure of nonoperative measures may necessitate surgery. Ideally the procedure should involve repairing the dural defect, removing the encapsulated cavity of the pseudomeningocele, and obliterating the extraspinal dead space to minimize the recurrence of the problem. METHODS: The authors describe a technique performed in 12 patients with large (> 5-cm-diameter) pseudomeningoceles referred for management following the failure of less aggressive measures. Diagnosis was based on symptoms of lumbar wound swelling, postural headaches, back and leg pain, and was confirmed by imaging studies. In all patients subarachnoid CSF drainage and initial operative attempts to obliterate the pseudomeningocele had failed. They were treated between July 1990 and July 1998. The cause of the pseudomeningoceles was lumbar discectomy (four patients), lumbar decompression (one patient), lumbar decompression and placement of instrumentation (five patients), and intradural procedures (two patients). Their mean age was 47.9 years (range 20-67 years), and they presented at a mean of 5.5 months postoperatively (range 3 weeks-37 months). In all cases there was a satisfactory repair of the pseudomeningocele, dead space obliteration, and long-term symptomatic resolution. CONCLUSIONS: Lumbar myofascial advancement for this problem is a useful technique in cases of symptomatic pseudomeningoceles. This technique requires the medial advancement of the musculofascial units of the paravertebral muscles for a layered closure over the exposed spinal canal with obliteration of the pseudomeningocele. PMID- 15347232 TI - Postoperative wound infections of the spine. AB - Postoperative spinal wound infections occur in 1 to 12% of patients. The rate of infection is related to the type and duration of the procedure, comorbidities, nutritional status, and various other risk factors. Antibiotic prophylactic therapy has been clearly shown to decrease the rate of infection dramatically after lumbar surgery. These infections typically manifest with signs and symptoms of wound swelling, erythema, and drainage. Laboratory-detected values such as the erythrocyte sedimentation rate and C-reactive protein can be elevated beyond what is normal for the uncomplicated postoperative course following lumbar surgery, and combined with the clinical symptoms should alert the physician to the possibility of infection. When detected, these infections should be managed aggressively with operative debridment and irrigation, including the deep subfascial layer in all cases except those with clearly demarcated superficial infection. The choice of one versus multiple debridments can be made based on the appearance of the wound, patient factors, and nutritional status. Hardware and incorporated bone graft can be left in place in the majority of cases, adding to stability. Outcomes following aggressive treatment of this complication can be excellent, with no long-term loss of function and complete eradication of the infection. PMID- 15347233 TI - Is spinal instrumentation a risk factor for late-onset infection in cases of distant infection or surgery? Case report. AB - As a cause of revision spinal surgery, spinal epidural abscess after instrumentation-assisted fusion is rare in neurosurgical practice. Postoperative infections are frequently seen in the time period soon after surgery. The authors report on the case of a 45-year-old woman who had undergone posterior instrumentation-augmented fusion for L4-5 degenerative spondylolisthesis. Ten months after the operation she presented to the neurosurgery clinic with complaints of severe low-back pain and radicular right lower-extremity pain. She had undergone laparoscopic surgery for acute cholecystitis 1 month prior to readmission. Radiological study revealed a spinal epidural abscess in communication with a right psoas abscess at L4-5. The abscess was drained percutaneously with the aid of C-arm fluoroscopic guidance, and a 6-week course of parenteral antibiotic therapy was administered. Retrograde lymphatic bacterial translocation, hematopoietic spread, and the suitable characteristics in the host may facilitate the development of infection around the implant. Thus, distant surgery and infection may be a risk factor in cases in which spinal instrumentation is placed. In such cases a prolonged antibiotic therapy for distant infection after surgery is recommended. PMID- 15347234 TI - [Is attention to mental health of the elderly with advanced disease an alternative function of internal medicine?]. PMID- 15347235 TI - [Hospital mortality in an Internal Medicine service]. AB - The Internal Medicine service of the Hospital General de Vic (Barcelona) takes part in the mortality committee by revising and discussing in-hospital mortality. BACKGROUND: to establish the characteristics of the deceased, death causes and to revise possible changes in the last six-years time or problems related to the exitus, to evaluate and improve hospitalized patients assistance. METHODOLOGY: Every case was revised following a specific register: demographical data, diagnosis and death cause, hospital death, documentation data, terminal or agonic situation when hospitalized, autopsies and death quality data. Exitus due to hospital problems were analyzed and classified in different groups. The statistical analysis was performed with measures of central tendency and of standard deviation. RESULTS: During the revised six years, there were 819 exitus (5.1%). Global average death age was 79 +/- 1.8 years: 52.5% were men and 47.4% were women; 22.8% died in less than forty-eight hours after hospitalization. The most frequent death causes were cerebrovascular accident (24%), chronic obstructive pulmonary disease (14.4%) and pneumonia (9.6%). There were a small number of autopsies (4.8%). Ratio of exitus due to hospital problems was stable during the six years (0.5%), in which nosocomial infection was the severest problem. CONCLUSIONS: The total percentage of exitus was 5.1%, higher than the common standards. Mortality causes coincide with other series. Ratio of exitus due to hospital problems was according to recommended objectives. The number of autopsies was very small. A correct completing and revision of the clinical recording is indispensable to spot a shortage in the hospitalized patients assistance. PMID- 15347236 TI - [Plasma lipoprotein (a) values in familial defective ligand apo B 100 in a South European population]. AB - AIMS: 1) to study lipoprotein (a) (Lp(a)) plasma values in subjects with familial ligand-defective apo B 100 (FDB). METHODS: We studied 19 heterozygous FDB subjects (8 males) from 12 families, carriers of R3500Q mutation on apo B gene and 90 controls (34 males). The genetic diagnosis was established with PCR-SSCP analysis and automatic sequencing. In all subjects plasma lipids, apolipoprotein B and Lp(a) levels were determined with standard procedures. RESULTS: Subjects carriers of R3500Q mutation on apo B gene have significantly higher plasma Lp(a) and log transformed Lp(a) values and prevalence of Lp(a) > 30 cut point for coronary heart disease than controls. CONCLUSIONS: Subjects with FDB showed higher Lp(a) plasma values than controls, although the mechanism and the clinical consequences of these result are not known. PMID- 15347237 TI - [Pulmonary embolism in medical inpatients. An approach to trends and costs in Spain]. AB - BACKGROUND: Tromboprophylaxis for hopitalized patients has became a widespread practice in the lasts years. However, the trends in pulmonary embolism (EP) are insufficiently known. We analyzed the trends of EP during the 1997-1999 period in Spain at the hospitals of the National Health System (NHS) and theirs costs were also estimated. METHODS: Diagnosis-Related Group (DGR) 78 from the data of the national hospital discharge register was evaluated. Annual trends, age, sex and (for 1999) mortality, some comorbidities and costs according official data from the Spanish Ministry of Health were annotated. Mostly medical patients were included. Data were compared with those from the Servicio Gallego de Saude (SERGAS) which maintain a high rate of hospitalary discharges declaration. RESULTS: During the 3-year period 14.021 cases of EP were observed. The 47% of the NHS cases were older than 75 and mortality was 6,8%. An annual increment of hospitalized cases was observed at SERGAS and NHS (in the last 5% at the end of the period). In 1999 costs were estimated in 16-20,2 millions euro. That was 0,097-0,12% of global hospitalary budget. CONCLUSIONS: Trends for hospitalizations for EP are increasing in Spain. It remains in doubt if a true increase in incidence or the improving notification and awareness are the responsibles for this increment. However, the data of SERGAS support the first possibility. PMID- 15347238 TI - [Tuberculous peritonitis: a Third World's disease?]. AB - Tuberculous peritonitis has a high morbidity and mortality. The clinical outcome of tuberculous peritonitis depends on an early diagnosis and treatment. We review five cases of tuberculous peritonitis diagnosed in a short stay medical unit over a one-year period. All patients were successfully treated with antituberculous drugs. The clinical management of this disease is described and current views regarding the value of computed tomography and other diagnostic techniques are discussed. PMID- 15347239 TI - [One case of pneumonia with Arcanobacterium pyogenes]. AB - Arcanobacterium pyogenes is an anaerobic Gram-positive bacilli and it is a part of the normal flora in many domestic animals. A. pyogenes is a rare cause of pyogenic infections in humans and most of cases reported are questionable since there was failure to definitively identify the pathogen. A man with no past medical history presented with respiratory infection. The blood sample collected confirmed an Arcanobacterium pyogenes. It was necessary the addition of clarytromycin to the initial empirical treatment with cefotaxime, to the complete recovery. This case provide further evidence that A. pyogenes can be a pathogen in humans even in absence of predisposing illness. PMID- 15347240 TI - [Psoas muscle abscess by Streptococcus agalactiae as a of presentation of fever of unknown origin]. AB - We present the case of a patient with precedents of endometrial carcinoma treated by surgery and radiotherapy, which presented a psoas abscess by Streptococcus agalactiae as cause of fever of unknown origin, which answered favorably to the antibiotherapy and surgical drainage. PMID- 15347241 TI - [Autoimmune hepatitis]. AB - Autoimmune hepatitis (AIH) is a hepatocellular inflammation that is characterised by a wide range of histopathologic (periportal interface hepatitis with plasma cell infiltration and piecemeal necrosis), biochemical (hypertransaminasemia, hypergammaglobulinaemia) and autoimmune (several autoantibodies presence) features. This relatively rare disorder frequently affects middle-aged women. There is no pathognomonic marker for AIH diagnosis, therefore it requires a careful rule out of other causes of liver disease together with the detection of a suggestive pattern of clinical and laboratory abnormalities. Scoring system for AIH diagnosis proposed by International Autoimmune Hepatitis Group has been used as a tool in clinical practice but is not sufficiently exclusive in terms of defining prognosis or treatment. AIH has been classified in two subtypes according to autoantibodies detected: 1 and 2, but this classification results in poor clinical implications. Previously known as subtype 3 is at the present included in subtype 1 because no clinical significant differences has been found between them. Aetiology, and molecular mechanisms still remain to be elucitaded in this disease, although viruses, drugs and molecular mimicry act presumably as a trigger in genetically predisposed patients (associated with HLA-DR3 and DR4 haplotypes). On the other hand, immunosuppressive therapy (corticosteroid or azathioprine) generally offers favourable response. Our aim is to review this disease from different points of view, considering: clinical, histopathological, etiologic, genetic, biochemical, autoimmune, treatment and prognosis features. PMID- 15347242 TI - [The different ways of dying. Ethical reflections]. AB - Death Process is seen influenced by multitude of factors. It's not habitual that the patient decides about the circumstances of his death. In the terminally ill situation, the Palliative Medicine is considered the best option. Sometimes the death is produced as consequence of an action or omission, which raises ethical important dilemmas difficult to resolve. The similarities and differences between active euthanasia and passive euthanasia are analysed. PMID- 15347243 TI - [Right hemiparesia as presentation of renal cell carcinoma]. PMID- 15347244 TI - [Cytomegalovirus infection mimicking a pseudomigraine with pleocytosis]. PMID- 15347245 TI - [Acute adrenal insufficiency and electrolytic alterations]. PMID- 15347246 TI - [Intestinal tuberculosis and pulmonary tuberculosis. Ileal resection and PCR for the diagnosis]. PMID- 15347247 TI - [Chest pain due to respiratory infection]. PMID- 15347248 TI - [Bilateral otitis media as presentation form of microscopic polyangiitis]. PMID- 15347249 TI - Proteomic patterns to identify ovarian cancer: 3 years on. PMID- 15347250 TI - Diagnostic applications of chromosomal translocations in bone and soft tissue sarcomas. PMID- 15347252 TI - OraQuick ADVANCE Rapid HIV-1/2 antibody test. AB - Rapid HIV antibody tests represent a key development in the current diagnosis and management of HIV infection. The OraQuick ADVANCE Rapid HIV-1/2 antibody test (OraSure Technologies) has received US Food and Drug Administration approval on the basis of its performance characteristics and a subsequent Clinical Laboratory Improvement Amendments waiver based on its simplicity and accuracy. The test has been approved for use on oral mucosal transudate, whole blood or plasma. Clinical evaluation of the OraQuick ADVANCE Rapid HIV-1/2 antibody test has revealed high sensitivity and specificity. The test has many important applications, extending the opportunities for voluntary counseling and testing, and as a tool for the scale-up of antiretroviral therapy in resource-limited settings. PMID- 15347253 TI - FIBROSpect II: a potential noninvasive test to assess hepatic fibrosis. AB - Many forms of liver disease may ultimately lead to fibrosis of the liver, the most advanced state being cirrhosis. Cirrhosis is a morphologic disease that eventually results in a functional change of the liver. It is generally not accompanied by signs or symptoms early in its course, and is often diagnosed late when signs of liver failure become overt. Imaging studies may suggest cirrhosis, but only if there have been gross changes in the appearance of the liver, and this is often not the case. The only way to diagnose cirrhosis reliably has been through direct histologic examination of liver tissue. The drawback to histologic diagnosis has been the risks and discomfort associated with liver biopsy. Hesitation to perform the procedure also exists due to lack of experience of many practitioners and the low reimbursement rates for a procedure that is viewed as time consuming and potentially dangerous. The search for noninvasive modalities to assess fibrosis through biochemical and other means has begun. Several markers are currently under investigation, many of which are combined with clinical assessment and other biochemical parameters, to establish the presence of liver fibrosis. FIBROSpect II is an example of a commercially available assay that employs a combination of three markers to distinguish between no, minimal and advanced fibrosis. PMID- 15347254 TI - Gene expression profiling for molecular staging and prognosis prediction in colorectal cancer. AB - A key issue for patients undergoing surgery for colorectal cancer is the accurate prediction of treatment outcome. Currently, classification of a tumor by histopathologic stage is the most accurate prognostic factor for the risk assessment of treatment failure. However, despite improved histologic techniques and the application of novel immunohistochemical and molecular techniques, it is still not possible to delineate the underlying biochemical and genetic events that predict clinical outcome for individual cancer patients. One reason for this lack of progress is that the factors which determine the metastatic potential of a primary tumor are still unknown. This reality, coupled to dramatic technological developments in the field of expression profiling, has started a paradigm shift in the staging of colorectal cancers. It has raised expectations that genetic and/or transcriptome profiling of the primary tumor will result in the identification of prognostic determinants relevant to the individual patient. In turn, this may allow a clinically relevant definition of patient subgroups based on individual molecular parameters for rational decision making regarding choice of therapy. PMID- 15347255 TI - Future potential of the Human Epigenome Project. AB - Deciphering the information encoded in the human genome is key for the further understanding of human biology, physiology and evolution. With the draft sequence of the human genome completed, elucidation of the epigenetic information layer of the human genome becomes accessible. Epigenetic mechanisms are mediated by either chemical modifications of the DNA itself or by modifications of proteins that are closely associated with DNA. Defects of the epigenetic regulation involved in processes such as imprinting, X chromosome inactivation, transcriptional control of genes, as well as mutations affecting DNA methylation enzymes, contribute fundamentally to the etiology of many human diseases. Headed by the Human Epigenome Consortium, the Human Epigenome Project is a joint effort by an international collaboration that aims to identify, catalog and interpret genome wide DNA methylation patterns of all human genes in all major tissues. Methylation variable positions are thought to reflect gene activity, tissue type and disease state, and are useful epigenetic markers revealing the dynamic state of the genome. Like single nucleotide polymorphisms, methylation variable positions will greatly advance our ability to elucidate and diagnose the molecular basis of human diseases. PMID- 15347256 TI - Breath analysis in critically ill patients: potential and limitations. AB - Breath tests are attractive since they are noninvasive and can be repeated frequently in the dynamically changing state of critically ill patients. Volatile organic compounds can be produced anywhere in the body and are transported via the bloodstream and exhaled through the lung. They can reflect physiologic or pathologic biochemical processes such as lipid peroxidation, liver disease, renal failure, allograft rejection, and dextrose or cholesterol metabolism. This review describes the diagnostic potential of endogenous organic volatile substances in the breath of critically ill patients. Since many of these patients require ventilatory support, aspects of breath analysis under mechanical ventilation will be addressed. Analytical procedures, problems concerning the physiologic meaning of breath markers and future developments will be discussed. PMID- 15347257 TI - Diagnosis and management of MELAS. AB - Mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) is the most common maternally inherited mitochondrial disease. An A-->G mutation in the transfer RNA(Leu(UUR)) gene at position 3243 of the mitochondrial DNA accounts for most MELAS cases. The transient nature of the stroke-like episodes is reflected in abnormalities on neuroimaging. The cardinal laboratory abnormalities include elevated serum lactate during the acute episodes and respiratory enzyme defects in skeletal muscle. Muscle biopsy also helps confirm the diagnosis by identifying abnormal proliferation of mitochondria. Although current treatment options for MELAS are largely supportive, several therapeutic approaches have been attempted with limited success. Genetic counseling is an important component of patient management in MELAS. Newer reproductive technologies hold promise for reducing the recurrence of MELAS in subsequent generations. Advances in research into gene therapy offer hope of treatment for the future. PMID- 15347258 TI - Exploring the genetic basis of disease using RNA interference. AB - Cancer and autoimmunity are polygenic diseases. In order to better understand the mechanisms of disease development and progression it is essential to uncover which genes are involved. Much has been learned from population studies in human patients by searching for polymorphic genetic loci associated with disease. In addition, animal models of tumor development, as well as models for various autoimmune conditions such as multiple sclerosis and Type I diabetes, have helped determine genetic loci that contribute to disease susceptibility. However, characterization of the exact genes involved is often difficult and requires lengthy and technically demanding genetic manipulations. The generation of knockout animals is the method of choice to probe single genes. However, this is not possible in all species or even in all inbred strains within a species. The recent discovery of a new post-transcriptional gene silencing pathway termed RNA interference, which is mediated by short fragments of double-stranded RNA (short interfering RNA), has opened up new avenues for genetic manipulation of experimental animals. This review will consider how silencing genes by RNA interference within the context of experimental disease models promises to become a powerful new tool for the genetic analysis of cancer and autoimmunity. Advances in RNA interference technology now permit the relatively rapid generation of transgenic animals in a wide range of species with complex genetic backgrounds that were previously inaccessible to genetic manipulation. This novel approach should help refine the characterization of disease-associated genes, either by silencing specific candidates or even by screening a larger number of genes in vivo within a comparatively short time frame. PMID- 15347259 TI - In situ hybridization: detecting viral nucleic acid in formalin-fixed, paraffin embedded tissue samples. AB - In situ hybridization is a method for detecting specific nucleic acid sequences within individual cells. This technique permits visualization of viral nucleic acid or gene expression in individual cells within their histologic context. In situ hybridization is based on the complementary binding of a labeled nucleic acid probe to complementary sequences in cells or tissue sections, followed by visualization of target sequences within the cells. It has been used widely for the detection of viral nucleic acid sequences within individual cells. This review will define the technical approaches of in situ hybridization and its current application to detect viral nucleic acids within formalin-fixed, paraffin embedded tissue samples, with special reference to the Epstein-Barr virus. PMID- 15347261 TI - Diagnosis of lysosomal storage disorders: current techniques and future directions. AB - Lysosomal storage disorders represent a group of over 45 distinct genetic diseases. The broad spectrum of clinical presentation of this group of disorders has led to the development of diagnostic protocols to facilitate their rapid and accurate diagnosis. However, with the development of new therapies, testing for many of these disorders now extends beyond diagnosis of affected individuals. The efficacy of many current and proposed therapies will rely heavily upon early detection and treatment prior to the onset of irreversible pathology. Newborn screening holds the promise of early detection. However, presymptomatic diagnosis raises a number of issues relating to patient management and treatment. Methods for prognoses and monitoring therapy in asymptomatic individuals will be required. PMID- 15347260 TI - Advanced molecular cytogenetics in human and mouse. AB - Fluorescence in situ hybridization, spectral karyotyping, multiplex fluorescence in situ hybridization, comparative genomic hybridization, and more recently array comparative genomic hybridization, represent advancements in the field of molecular cytogenetics. The application of these techniques for the analysis of specimens from humans, or mouse models of human diseases, enables one to reliably identify and characterize complex chromosomal rearrangements resulting in alterations of the genome. As each of these techniques has advantages and limitations, a comprehensive analysis of cytogenetic aberrations can be accomplished through the utilization of a combination approach. As such, analyses of specific tumor types have proven invaluable in the identification of new tumor specific chromosomal aberrations and imbalances (aneuploidy), as well as regions containing tumor-specific gene targets. Application of these techniques has already improved the classification of tumors into distinct categories, with the hope that this will lead to more tailored treatment strategies. These techniques, in particular the application of tumor-specific fluorescence in situ hybridization probes to interphase nuclei, are also powerful tools for the early identification of premalignant lesions. PMID- 15347262 TI - Surrogate electroretinographic markers for assessing therapeutic efficacy in the retina. AB - Visual acuity remains the primary surrogate marker for clinical trials in ophthalmology (and the primary outcome for most US Food and Drug Administration applications) due to its long history, ease of measurement and clear relationship to clinically meaningful characteristics of daily life. However, treatment trials are being planned for diseases that are currently untreatable where visual acuity may not be the most appropriate outcome measure. Specialized electroretinographic tests can be powerful surrogate markers in such trials. The selection of outcome measures and surrogate markers depends in part on whether the goal is to preserve remaining vision in a progressive retinal degeneration or to improve vision in an eye that has already undergone extensive degeneration. Among the electroretinographic tests available are those involving the whole retina (full field electroretinographic), the posterior pole (pattern electroretinographic) or focal areas within the macula (multifocal electroretinographic). The advantages and disadvantages of each will be discussed along with selected applications of each test to a specific category of disease. PMID- 15347263 TI - Assessment of drug resistance in acute myeloid leukemia. AB - A major problem in the treatment of leukemia is the development of resistance to chemotherapeutic agents. Assessing the drug resistance of leukemic cells is therefore an important aspect of treatment. One of the main mechanisms of resistance is rapid drug efflux mediated by various members of the ATP-binding cassette transporter superfamily, such as multidrug resistance gene 1 (MDR1), which encodes P-glycoprotein, multidrug resistance-associated protein (MRP) 1 and lung resistance protein. To quantify the degree of acquisition of resistance, several techniques, including drug-sensitivity studies, flow cytometry assay and quantitative gene analysis, have been developed to detect MDR1 and MRP1 gene expression in leukemic cells. However, a significant number of patients may relapse in spite of low expression of MDR1 or MRP1, suggesting the involvement of other intracellular mechanisms, possibly related to cytarabine resistance. This review focuses on the methods aimed at the assessment of drug resistance in acute myeloid leukemia. PMID- 15347264 TI - Noninvasive diagnosis of fibrosis in chronic liver disease. AB - Laboratory tests for the noninvasive diagnosis of liver fibrosis were studied extensively in the past. However, no test is yet accepted to replace liver biopsy as the gold standard. The establishment of widely accepted semiquantitative histologic scoring systems for the grading and staging of chronic liver disease (e.g., Ishak, Metavir and Scheuer) was paralleled by a significant upturn of research in circulating markers of liver fibrosis. We are now experiencing the renaissance of standard clinical chemistry markers, which are assembled to multiparameter scores (e.g., aspartate aminotransferase-to-platelet ratio index, FibroTest, Forns' index). These scores still require comprehensive evaluation in comparison with histology. Better understanding of the pathophysiology of liver fibrosis provided new options regarding circulating markers of hepatic matrix metabolism (e.g., hyaluronic acid, laminin, matrix metalloproteinase-2, aminoterminal propeptide of procollagen type III and tissue inhibitors of metalloproteinases-1). Several promising studies have been published to date. Thus, a redefinition of the role of liver biopsy is expected in the foreseeable future. PMID- 15347265 TI - RT-PCR tyrosinase expression in the peripheral blood of melanoma patients. AB - Controversial data are reported in the literature concerning the role of peripheral blood tyrosinase messenger RNA reverse transcription PCR analysis in melanoma patient management. Some papers assess the clinical relationship between tyrosinase expression and disease outcome, while others address the high degree of variability in the positive rate percentage and demonstrate the transient shedding of melanoma cells in the bloodstream. An overview of the current knowledge about the applications and limitations of tyrosinase analysis compared with other biologic markers is presented herein. Tyrosinase expression should not be considered as a tumor burden-related marker in the peripheral blood, but rather as a measure of the potential increased risk of metastatic spreading. In this view, reverse transcription PCR gains a clinical significance when sequential determinations are performed during follow-up, whereas the evaluation of a single sample, either positive or negative, does not bring any additional clinical information. PMID- 15347266 TI - Passive smoking and breast cancer: is the evidence for cause now convincing? PMID- 15347267 TI - The SAFE study: a landmark trial of the safety of albumin in intensive care. AB - High-quality primary evidence from an Australian and New Zealand study provides a definitive answer. PMID- 15347268 TI - Suspected child abuse: false positives or false negatives? PMID- 15347269 TI - Reducing drug-related harm: Australia leads the way. AB - Harm-reduction approaches are more easily embedded in policy when drugs are legally regulated. PMID- 15347270 TI - Risk factors for ischaemic stroke recurrence after hospitalisation. AB - OBJECTIVE: To determine risk factors for ischaemic stroke recurrence among patients admitted to hospital for a first-ever occurrence of ischaemic stroke. DESIGN, SETTING AND PATIENTS: Retrospective study involving linked hospitalisation and death records. The cohort comprised 7816 people who were hospitalised for first-ever ischaemic stroke between July 1995 and December 1999 in Western Australia. Cox's proportional hazards model was used to identify risk factors for stroke recurrence. MAIN OUTCOME MEASURES: Time to first recurrence; cumulative recurrence risk; risk factors for recurrence. RESULTS: The median time to first stroke recurrence was 255 days. The cumulative probability of first recurrence was 5.1% (95% CI, 4.6%-5.7%) at 6 months, 8.4% (95% CI, 7.6%-9.1%) at 1 year and 19.8% (95% CI, 18.1%-21.4%) at 4 years. The risk of first recurrence was increased by advancing age (hazard ratio [HR], 1.03; 95% CI, 1.02-1.04), Aboriginality (HR, 1.50; 95% CI, 1.02-2.22), diabetes (HR, 1.27; 95% CI, 1.07 1.51), a history of cardiac conditions (HR, 1.18; 95% CI, 1.01-1.38), post-stroke urinary incontinence (HR, 1.27; 95% CI, 1.03-1.57) and transfer to another hospital on index admission (HR, 1.26; 95% CI, 1.08-1.46). Admission at first stroke occurrence to a hospital maintaining a stroke unit reduced the risk of recurrence (HR, 0.84; 95% CI, 0.72-0.99). CONCLUSION: The risk factors identified in our study have implications for planning secondary prevention strategies. In particular, Aboriginality and transfer to another hospital upon admission for first-ever ischaemic stroke were important risk factors. Research into the level of compliance and access to stroke treatment by Aboriginal patients to prevent further strokes is required. PMID- 15347271 TI - Use of complementary and alternative medicines by patients with chronic obstructive pulmonary disease. AB - OBJECTIVES: To investigate complementary and alternative medicine (CAM) use by patients with chronic obstructive pulmonary disease (COPD) and to explore their beliefs about CAM. DESIGN AND PARTICIPANTS: Cross-sectional study of 173 patients with moderate to severe COPD, and in depth interviews with a purposive sample of 28 patients. SETTING: Ambulatory care. MAIN OUTCOME MEASURES: Use of CAM; beliefs about the value of CAM. RESULTS: 71 patients (41%) claimed to be using some form of CAM. Most commonly used were multivitamins and minerals, and garlic was the most commonly used herbal preparation. Patients reported that advertisements and people with prior experience of using CAM were their major sources of information. Extent of knowledge about CAM, degree of faith in CAM and personal attitudes influenced decisions to try CAM. Patients used CAM to promote general wellbeing, to counteract drug side effects, to compensate for dietary deficiencies and to ameliorate their disease. Efficacy appeared less important to users than safety. CAM practitioners were regarded as more convincing, informative, considerate and available compared with mainstream health professionals. CONCLUSIONS: Communication between patients and mainstream health professionals about CAM use could be improved by health professionals being more accepting of CAM use and having some basic knowledge about commonly used CAM preparations. PMID- 15347272 TI - The potential for tobacco control to reduce PBS costs for smoking-related cardiovascular disease. AB - OBJECTIVE: To estimate Pharmaceutical Benefits Scheme (PBS) subsidies for drugs to treat smoking-related cardiovascular disease (CVD) in 2001-02, and over the period of the government's Intergenerational Report (IGR), assuming current smoking prevalence rates and a 5% absolute reduction. DESIGN AND SETTING: An Australian epidemiological study, using prescribing data, aetiological fraction methodology, and IGR trends. MAIN OUTCOME MEASURES: Estimated smoking-related PBS subsidy costs in 2001-02 and predicted cumulative subsidies until 2041-42, under current and reduced smoking prevalence assumptions. RESULTS: The PBS costs of smoking-related CVD in 2001-02 were $126 million, 9.77% of the cost of drugs for CVD and 2.96% of total PBS subsidies. The cumulative difference in these costs over the 40-year period with a 5% drop in smoking prevalence was predicted to be $4.5 billion, a 17% reduction. The saving would be $1.14 billion discounting future costs at 5% per year. CONCLUSIONS: Further investment in tobacco control interventions could curb the increasing cost of the PBS and contribute to government efforts to ensure the viability of Australia's healthcare-financing programs. The net present value of a campaign to reduce smoking prevalence was estimated at $1 billion, with an internal rate of return of 33%. PMID- 15347273 TI - Will the Australia-United States Free Trade Agreement undermine the Pharmaceutical Benefits Scheme? AB - The Australia-United States Free Trade Agreement (AUSFTA) contains major concessions to the US pharmaceutical industry that may undermine the egalitarian principles and operation of the Pharmaceutical Benefits Scheme (PBS) and substantially increase the costs of medicinal drugs to Australian consumers. AUSFTA's approach to the PBS excessively emphasises the need to reward manufacturers of "innovative" new pharmaceuticals, instead of emphasising consumers' need for equitable and affordable access to necessary medicines (the first principle of our National Medicines Policy). Several features of AUSFTA may bring pressure to bear on the Pharmaceutical Benefits Advisory Committee (PBAC) to list "innovative" drugs that the committee initially rejected because the evidence for cost-effectiveness was not compelling. Intellectual property provisions of AUSFTA are likely to delay the entry of PBS cost-reducing generic products when pharmaceutical patents expire. We support the many concerned health and consumer organisations who have asked the Senate either not to pass the enabling legislation, or to delay its passage until a fairer deal in terms of public health can be obtained. PMID- 15347274 TI - Free trade in pharmaceuticals. AB - Provisions in the Australia-United States Free Trade Agreement (AUSFTA) may threaten the Australian Pharmaceutical Benefits Scheme (PBS), the "gold standard" of such programs worldwide. If Australia postpones passing of the US Free Trade Agreement Implementation Bill in the Senate, there will be opportunity for broader interests in both the United States and Australia to carefully study the agreement. The provisions of AUSFTA relating to the PBS are supposed to promote transparency, but the pharmaceutical manufacturers themselves (who are demanding transparency) do not reveal the content of their submissions to the Pharmaceutical Benefits Advisory Committee, or disclose all their financial relationships with researchers and policymakers. In AUSFTA, the "public health" language of affordable prescription drugs is missing and is replaced by language supporting "pharmaceutical innovation". Debate as to whether AUSFTA will force significant changes to the PBS, including higher drug prices, is currently under way in Australia. Perhaps the appropriate target of reforms should be the excessive US drug prices, and not the economically efficient Australian drug prices. PMID- 15347275 TI - Paget's disease of bone. AB - Paget's disease of bone is common, affecting up to 4% of Australians over the age of 55 years. The incidence of the disease and the severity of newly diagnosed cases appear to be falling, for unknown reasons. The cause of Paget's disease is unknown, but there is a strong genetic influence. Recently, mutations in the sequestosome 1/p62 gene have been identified as a cause of familial Paget's disease and of some apparently sporadic cases of the disease. The disease is often asymptomatic, but can cause bone pain, deformity, fracture and other complications. Paget's disease is eminently treatable. Potent bisphosphonates such as pamidronate, alendronate and risedronate relieve symptoms and may reduce the risk of complications. The Pharmaceutical Benefits Scheme subsidises treatment only for patients with symptomatic disease. A strong case be made for also treating asymptomatic patients with involvement of long bones, vertebrae or base of skull, patients with significant osteolytic lesions, and perhaps all younger patients. PMID- 15347276 TI - Public reporting of individual surgeon performance information: United Kingdom developments and Australian issues. AB - The United Kingdom is currently introducing public reporting of performance information for individual cardiac surgeons. The reports will indicate whether a surgeon has an acceptable level of performance, measured by in-hospital mortality. In the United States, surgeon-specific performance data have been available for over a decade. Arguments from both safety and accountability perspectives provide strong justifications for public reporting of such data. Were Australia to adopt similar public reporting processes, we should learn from overseas experiences. Surgical associations should be actively involved in developing data standards and processes for data collection, validation, analysis and publication. Any Australian policy initiative for public reporting of individual surgeon data should be backed by a political commitment to adequate funding. PMID- 15347277 TI - Japanese encephalitis vaccine: is it being sufficiently used in travellers? PMID- 15347278 TI - Time for hard decisions on patient-centred professionalism. AB - Patients want doctors who are competent, respectful, honest and able to communicate with them. That is patient-centred professionalism. Professional self regulation, as practised hitherto, has failed to achieve this for all patients. In the United Kingdom, a new way of looking at professional regulation has been developed--as a partnership between public and doctors. At its heart is a code of good practice, agreed between public and profession, in which doctors' licence to practise is conditional on regularly demonstrating continuing compliance. That means revalidation-relicensure. PMID- 15347279 TI - Undergraduate teaching of complementary medicine. PMID- 15347280 TI - Teaching integrated care: CAM familiarisation courses. AB - As the use of CAM grows, CAM familiarisation offers educational opportunities for undergraduates to understand CAM, their attitudes to medical change and the process of evidence-based medicine. Such courses also offer the opportunity to integrate patient care and improve the relationship between medical and CAM practitioners. CAM familiarisation courses are available in many medical schools in the United States and the United Kingdom. The multiprofessional model which we have developed at the University of Southampton (UK) offers valuable experience to those thinking of establishing such courses. PMID- 15347281 TI - Japanese encephalitis acquired near Port Moresby: implications for residents and travellers to Papua New Guinea. PMID- 15347282 TI - New recommendation on Japanese encephalitis vaccination for travellers to Papua New Guinea. PMID- 15347283 TI - Smoking cessation and elective surgery: the cleanest cut. PMID- 15347284 TI - Smoking and pregnancy. PMID- 15347285 TI - Androgen deficiency and replacement therapy in men. PMID- 15347286 TI - Treatment of osteoporosis: why, whom, when and how to treat. PMID- 15347287 TI - Unexpected infant death: lessons from the Sally Clark case. PMID- 15347288 TI - Identification, molecular cloning and functional characterization of a novel NADH kinase from Arabidopsis thaliana (thale cress). AB - NADH kinase (NADHK; ATP:NADH 2'-phosphotransferase; EC 2.7.1.86), an enzyme that preferentially utilizes NADH as the diphosphonicotinamide nucleotide donor, has been identified for the first time in plants. Low activity (0.4 nmol of NADPH produced/min per mg of protein) was observed in clarified protein extracts from Arabidopsis thaliana (thale cress) cell suspension cultures. However, unlike an NADHK from yeast (Saccharomyces cerevisiae) (POS5), the enzyme from Arabidopsis did not associate with the mitochondria. NADHK was cloned (gi:30699338) from Arabidopsis and studied as a recombinant protein following affinity purification from Escherichia coli. The enzyme had a pH optimum for activity of 7.9 and a subunit molecular mass of 35 kDa. Analytical gel filtration demonstrated that the recombinant enzyme exists as a dimer. Hyperbolic saturation kinetics were observed for the binding of NADH, ATP, free Mg2+ and NAD+, with respective K(m) values of 0.042, 0.062, 1.16, and 2.39 mM. While NADHK could phosphorylate NADH or NAD+, the specificity constant (V(max)/K(m)) for NADH was 100-fold greater than for NAD+. The enzyme could utilize UTP, GTP and CTP as alternative nucleotides, although ATP was the preferred substrate. PP(i) or poly-P(i) could not substitute as phospho donors. PP(i) acted as a mixed inhibitor with respect to both NADH and ATP. NADHK was inactivated by thiol-modifying reagents, with inactivation being decreased in the presence of NADH or ATP, but not NAD+. This study suggests that, in Arabidopsis, NADP+/NADPH biosynthetic capacity could, under some circumstances, become uncoupled from the redox status of the diphosphonicotinamide nucleotide pool. PMID- 15347289 TI - Effect of obturation technique on sealer cement thickness and dentinal tubule penetration. AB - AIM: To compare the average sealer cement film thickness and the extent and pattern of sealer penetration into dentinal tubules in association with four obturation techniques in curved root canals. METHODOLOGY: Mesial canals of 44 extracted mandibular molars were randomly divided among the SimpliFill, continuous wave, Thermafil and 0.04 matched taper (master cones) lateral compaction obturation groups (22 canals per group). AH26 sealer cement was coloured blue-black using Sudan Black B dye. Roots were sectioned 1, 3 and 5 mm from the working length. Specimens were photographed under 25x magnification, mounted as 35 mm slides and projected. Average sealer cement thickness (measured at 10 points around the canal wall), depth of dentinal tubule penetration and frequency of voids were determined at the 1, 3 and 5 mm levels. Data were analysed statistically for effect of obturation technique and level of section on sealer thickness and on the depth and distribution of tubule penetration. RESULTS: Thermafil demonstrated superior GP adaptation at all levels with a mean overall sealer cement thickness of 2.2 microm, followed by lateral compaction (11.1 microm), continuous wave (12.2 microm) and SimpliFill (47.6 microm). SimpliFill also demonstrated the highest frequency of voids (P < 0.05). Sealer cement penetrated dentinal tubules as far as the outer one-third of dentine, with greater penetration observed buccally or lingually (P < 0.001). Penetration was not significantly affected by obturation technique, but on average was deeper and more frequent at the 3 and 5 mm levels than at the 1 mm level (P < 0.001). CONCLUSIONS: Sealer thickness was strongly dependent on obturation technique. Assuming that minimal sealer thickness and fewer voids are good measures of long term sealing ability, Thermafil resulted in the best outcome. Consistent, extensive sealer penetration into dentinal tubules was seen and was unrelated to the obturation technique. PMID- 15347290 TI - An in-vitro investigation of the antibacterial effect of nisin in root canals and canal wall radicular dentine. AB - AIM: To determine whether nisin, a bacteriocin, would be effective at killing Enterococcus faecalis and Streptococcus gordonii cells in solution and within the root canal system. METHODOLOGY: Bacterial isolates of E. faecalis and S. gordonii were grown from glycerol stocks in closed tubes containing BHY broth at 37 degrees C. The minimum bactericidal concentration (MBC) of nisin for both bacterial species was determined by a microdilution method. Extracted human teeth were decoronated to produce roots of equal length with a single canal and divided into six groups of 10 roots. The canals were prepared to a master apical size 30 file using 0.04 taper Ni-Ti rotary instruments. Bacterial samples of each species were inoculated into three groups of prepared roots and incubated in closed tubes at 37 degrees C for 21 days. The root canals in each group were then medicated with water (control), calcium hydroxide powder mixed with sterile water [Ca(OH)2], or nisin and incubated for a further 7 days. Rotary Ni-Ti files were used to take radicular dentine samples from the walls of each canal which were then incubated in BHY broth for 24 h. Optical density (OD600) readings were taken as a measure of bacterial growth. RESULTS: The MBC of nisin for E. faecalis and S. gordonii was 70 and 20 mg mL(-1) respectively. Calcium hydroxide and nisin medication eradicated infection within the root canal while cells remained viable in the control group. Mean optical density (OD600) readings from canal wall dentine shavings infected with E. faecalis were 1.32 +/- 0.98, 0.73 +/- 0.27 and 0.69 +/- 0.38 for the control, Ca(OH)2 and nisin samples respectively. Corresponding mean readings for S. gordonii were 1.19 +/- 0.18, 0.73 +/- 0.15 and 0.60 +/- 0.29. The Ca(OH)2 and nisin group readings were significantly (P < 0.01) lower than the control for each species as tested by Student's t-test and Mann Whitney U statistical analysis. Values for Ca(OH)2 and nisin were not significantly (P > 0.01) different. CONCLUSION: Nisin was effective at eradicating E. faecalis and S. gordonii cells in pure culture and was comparable with Ca(OH)2 in the elimination of these species from within the root canal system. PMID- 15347291 TI - The effectiveness of syringe irrigation and ultrasonics to remove debris from simulated irregularities within prepared root canal walls. AB - AIM: To compare the ability of syringe irrigation and ultrasonic irrigation to remove artificially placed dentine debris from simulated canal irregularities within prepared root canals. METHODOLOGY: After canal enlargement, twelve canines were split longitudinally into two halves. On the wall of one half of each root canal a standard groove of 4 mm in length, 0.2 mm in width and 0.5 mm in depth was cut, 2-6 mm from the apex, to simulate uninstrumented canal extensions. On the wall of the other half, three standard saucer-shaped depressions of 0.3 mm in diameter and 0.5 mm in depth were cut at 2, 4 and 6 mm from the apex to simulate uninstrumented canal irregularities. Each groove and depression were filled with dentine debris mixed with 2% NaOCl to simulate a situation when dentine debris accumulates in uninstrumented canal extensions and irregularities during canal preparation. Each tooth was re-assembled by reconnecting the two halves, using wire and an impression putty material. Two per cent NaOCl was then delivered into each canal either using syringe irrigation (n = 8) or using ultrasonic irrigation (n = 8). Before and after irrigation, images of the two halves of the canal wall were taken, using a microscope and a digital camera, after which they were scanned into a PC as TIFF images. The amount of remaining dentine debris in the grooves and depressions was evaluated by using a scoring system between 0-3: the higher the score, the more the debris. The data were analysed by means of the Mann-Whitney U-test. RESULTS: Both forms of irrigation reduced the debris score significantly. The debris score was statistically significantly lower after ultrasonic irrigation than after syringe irrigation (P = 0.002 for grooves, P = 0.047 for depressions). CONCLUSION: Ultrasonic irrigation ex vivo is more effective than syringe irrigation in removing artificially created dentine debris placed in simulated uninstrumented extensions and irregularities in straight, wide root canals. PMID- 15347292 TI - A one step procedure for luting glass fibre posts: an SEM evaluation. AB - AIM: To evaluate the ability of two luting procedures for bonding translucent fibre posts to form resin tags, adhesive lateral branches, and resin dentine interdiffusion zones (RDIZ). METHODOLOGY: Forty root filled teeth, extracted for periodontal reasons, were selected for the study. The teeth were randomly divided into four groups of 10 each. Group 1: One Step, light-cured (LC) 20 s + dual link resin cement (LC 20 s); group 2: One Step, not light-cured (NLC) + dual link resin cement (LC 30 s); group 3: One Step (NLC) + dual link resin cement (LC 60 s); group 4: One Step (NLC) + dual link resin cement (LC 90 sec). Forty translucent fibre posts (DT posts, RTD) were inserted. Then, root specimens were processed for scanning electron microscope (SEM) observations to assess the continuity of the RDIZ, the presence or absence of gaps and the density and morphology of resin tags using a four-step scale method. RESULTS: With all luting procedures the formation of a RDIZ occurred. However, the microscopic examination of adhesive interfaces revealed that the percentage of RDIZ was significantly higher in group 1 than in the other three groups (P < 0.05). Group 4 had a higher percentage of RDIZ than group 3, which in turn exhibited a greater percentage than group 2, but these differences were not statistically significant (P > 0.05). Resin tag formation was evident in all the groups. The characteristic reverse cone shape of resin tags was always noted in the coronal and middle third of all groups, and in the apical third of group 1. In the apical third a shorter length and a less uniform appearance of resin tags were noted in groups 2, 3 and 4. CONCLUSIONS: In this laboratory study, when bonding a translucent fibre post into a root canal, the luting procedure including light curing of the adhesive solution before the placement of the cement was more satisfactory, from a SEM standpoint, than the procedure involving the simultaneous curing of adhesive and cement (one step procedure). PMID- 15347293 TI - Comparison of long-term papilla healing following sulcular full thickness flap and papilla base flap in endodontic surgery. AB - AIM: To compare long-term loss of papilla height when using either the papilla base incision (PBI) or the standard papilla mobilization incision in marginal full thickness flap procedures in cases with no evidence of marginal periodontitis. METHODOLOGY: Twelve healthy patients, free of periodontal disease, who had intact interdental papillae were referred for surgical treatment of persisting apical periodontitis and included in the study. The flap design consisted of two releasing incisions connected by a horizontal incision. The marginal incision involved the complete mobilization of the entire papilla in one interproximal space but in the other interproximal space the PBI was performed. Further apically a full thickness flap was raised. Following flap retraction, standard apical root-end resection and root-end filling was performed. Flap closure was achieved with microsurgical sutures. The PBI was sutured with two to three interrupted sutures (size 7/0), the elevated papilla was reapproximated with vertical mattress sutures (size 7/0), which were removed 3-5 days after the surgery. The height of the interdental papilla was evaluated preoperatively and postoperatively after 1-, 3- and 12-month recall using plaster replicas. The loss of papilla height was measured using a laser scanner. Papilla paired sites were evaluated and statistically analysed. RESULTS: Most papilla recession took place within the first month after the surgery in the complete elevation of the papilla. Further small increase in loss of papilla height resulted at 3 months. After 1 year the loss of height diminished to 0.98 +/- 0.75 mm, but there was no statistical difference between the various recall intervals. In contrast, after PBI only minor changes could be detected at all times. There was a highly significant difference between the two incision techniques for all recall appointments (P < 0.001). CONCLUSIONS: In the short as well as long-term the PBI allows predictable recession-free healing of the interdental papilla. In contrast, complete mobilization of the papilla displayed a marked loss of the papilla height in the initial healing phase although this was less evident 1 year postoperatively. In aesthetically relevant areas the use of the PBI is recommended, to avoid opening of the interproximal space, when periradicular surgical treatment is necessary. PMID- 15347294 TI - Root canal obturation by ultrasonic condensation of gutta-percha. Part II: an in vitro investigation of the quality of obturation. AB - AIM: To compare the quality of root canal obturation using ultrasonic or cold condensation of gutta-percha and to determine the effect of power setting and activation time on the quality of obturation using the former technique. METHODOLOGY: An extracted human maxillary canine was used in an in vitro split tooth model to allow repeated obturation of the same root canal system using an ultrasonic device to thermocompact gutta-percha without sealer. After each obturation, the root filling was removed from the tooth to allow evaluation of its quality and for the tooth to be re-obturated. The influence of combinations (n = 10 per combination) of power setting (1, 3, 5) and activation times (4, 10, 15 s) was tested on the quality of root filling, assessed by measuring the voids within the body of the root filling as well as at the surface. Image analysis was used to quantify the voids within the body of the root filling. Cold lateral condensation of gutta-percha served as a control. RESULTS: Both surface and cross sectional analyses revealed that different power setting and activation time combinations produced significantly fewer voids than cold lateral condensation (P < 0.05) at the apical, mid-root and coronal levels. CONCLUSIONS: Taking surface and cross-sectional analysis together only power setting 5 and activation times of 10 and 15 s consistently produced ultrasonically thermocompacted root canal fillings with fewer voids than cold lateral condensation without sealer. PMID- 15347295 TI - Biocompatibility of two commercial forms of mineral trioxide aggregate. AB - AIM: To examine the biocompatibility of two commercial forms of mineral trioxide aggregate (MTA), by evaluating the morphology of an established cell line. METHODOLOGY: The two cements were cast on glass cover slips and cured for 1 or 28 days. Saos-2 osteosarcoma cells were trypsinized and seeded at a density of 1 x 10(5) cells and were then placed in medium over the material-coated coverslips for 1, 5 and 7 days. After these time intervals the media were discarded and the cells fixed. Cell morphological investigation was performed by scanning electron microscopy at various magnifications ranging from x 250 to x 500. The biocompatibility of cement constituents, alusilicate flux and bismuth oxide was also investigated. RESULTS: All cement samples cured for 1 day showed a confluent cell monolayer after 5 and 7 days. The response to both materials was similar. Materials cured for 28 days showed incomplete cell confluence after 1 and 5 days. Alusilicate flux and bismuth oxide did not demonstrate biocompatibility. CONCLUSIONS: The 1-day cured samples of two commercial forms of MTA showed good biocompatibility. However, the 28-day cured samples were less biocompatible after 1 and 5 days. PMID- 15347296 TI - Susceptibility to localized corrosion of stainless steel and NiTi endodontic instruments in irrigating solutions. AB - AIM: To evaluate the pitting and crevice corrosion characteristics of stainless steel (SS) and NiTi endodontic files in R-EDTA and NaOCl irrigating solutions. METHODOLOGY: The corrosion behaviour of two H-files produced from different SS alloys (Mani, AISI 303 SS, Dentsply Maillefer, AISI 304 SS) and one file produced from NiTi alloy (Maillefer) was determined in R-EDTA and NaOCl irrigating solutions by the cyclic potentiodynamic polarization method. The cutting flutes of 12 files of each material were embedded in an epoxy resin, polished, exposed to the irrigating solutions and used as an electrode. An Ag/AgCl electrode was used as a reference, a platinum plate was used as a counter electrode and polarization curves were obtained for all files in R-EDTA and NaOCl irrigating solutions in 37 degrees C with a potential scan rate of 5 mV min(-1). Corrosion potential (Ecorr), Corrosion current density (Icorr) and Pitting potential (Epit) were calculated from each curve. The results were statistically analysed with two way anova and Student-Newman-Keuls (SNK) multiple comparison test with materials and irrigating solutions serving as discriminating variables (a = 0.05). RESULTS: Cyclic polarization curves presented negative hysteresis implying that pitting or crevice corrosion are not likely to occur for all the materials examined in both irrigating solutions. In NaOCl all materials showed significantly higher Ecorr (P = 0.011) as well as lower Icorr compared with R-EDTA reagent. Moreover, all materials demonstrated equal Epit in NaOCl, which was to be found significantly lower (P = 0.009) than the value of Epit in R-EDTA. CONCLUSIONS: None of the tested materials is susceptible to pitting or crevice corrosion in R-EDTA and NaOCl solutions and from this standpoint are appropriate for the production of endodontic files. PMID- 15347297 TI - Central adenoid cystic carcinoma of the mandible manifesting as an endodontic lesion. AB - AIM: To present a case of adenoid cystic carcinoma (ACC) in the mandible, and manifesting as a periapical lesion. SUMMARY: A 56-year-old male suffered from pain around the right mandibular first molar for approximately 1 week. Oral examination revealed that the involved tooth was restored by a full coverage crown with no obvious abnormalities. A periapical radiograph revealed two ill defined radiolucencies associated with the tooth, one over the mesial and another over the distal roots of the tooth; incomplete root filling and furcation involvement also being noted. The affected tooth was extracted based on the clinical impression of apical periodontitis. The surrounding tissue of the root apex was curetted and sent for histopathological examination, which revealed ACC. KEY LEARNING POINTS: Adenoid cystic carcinoma affecting the mandible may mimic a periapical lesion. Proper diagnosis of such a lesion is dependent on thorough clinical, radiographic and microscopic examinations. Such a case highlights the benefits of biopsy and histological examination of collected tissues. Diagnosis of lesions in the mandible should include salivary gland tumours. PMID- 15347298 TI - Endodontic treatment of an invaginated maxillary lateral incisor with a periradicular lesion and a healthy pulp. AB - AIM: To report the healing of a periradicular lesion following nonsurgical root canal treatment of a dental invagination. SUMMARY: A case of dens invaginatus type 3 with a periradicular lesion and a healthy pulp is reported. Nonsurgical root canal treatment of the invagination was performed successfully and resolved an associated periradicular lesion. Despite the complex anatomy and a diagnosis of apical periodontitis, pulp health was retained after endodontic treatment of the invaginated canal. Clinical considerations and treatment are discussed and reported. KEY LEARNING POINTS: Dens invaginatus can present in a variety of forms, knowledge of which can usefully inform endodontic diagnosis and treatment. In certain cases, success can be achieved by treating the invagination alone. Pulp health may be preserved during and after treatment of an infected invagination. PMID- 15347299 TI - Epidemiology of Helicobacter pylori infection. AB - This review summarizes key results of epidemiologic studies published in peer reviewed journals between April 2003 and March 2004. The prevalence of H. pylori infection continues to vary strongly between developing countries and developed countries, and according to ethnicity, place of birth and socioeconomic factors among people living in the same country. Intrafamilial spread appears to play a central role in transmission of the infection in both developing and developed countries. The role of H. pylori infection in development of noncardia gastric cancer appears to be even much stronger than previously assumed, whereas the lack of an association with cardia cancer and an inverse association with adenocarcinoma of the esophagus could be confirmed. Suggestions for an inverse association of the infection with atopic diseases have recently received further support, whereas evidence concerning the role of the infection (or its eradication) in GERD and a large variety of other extragastric diseases, including cardiovascular disease, remains inconclusive. PMID- 15347300 TI - Diagnosis of Helicobacter pylori infection. AB - While there are some attempts to improve culture of Helicobacter pylori, molecular methods have been the main focus of this interest. Their main application concerns the development of rapid tests also allowing the determination of bacterial resistance, i.e. real-time polymerase chain reaction (PCR) or fluorescence in situ hybridization (FISH), or to genotype the strains. Attempts to improve, simplify or explain the discrepancies of urea breath test results have been made and new generation of stool antigen test with monoclonal antibodies either using the standard ELISA format or rapid immunoenzymatic detection have confirmed their value. With regard to serology, studies have mainly focused on the distinction of infections with more pathogenic strains and the ability to diagnose atrophic gastritis with the Gastropanel. PMID- 15347301 TI - Pathogenesis of Helicobacter pylori infection. AB - Research in the last year has provided new insights into the function of the the cag-associated type IV secretion system and the vacuolating toxin VacA. A quite new aspect was disclosed by the finding that Helicobacter pylori in Mongolian gerbils colonizes a very distinct topology in the gastric mucous layer, obviously providing optimal conditions for long-term survival. Further research activities focused on H. pylori ammonia and metal metabolism as well as on bacterial stress defence mechanisms. Differential expression of approximately 7% of the bacterial genome was found at low pH suggesting that H. pylori has evolved a multitude of acid-adaptive mechanisms. VacA was shown to interrupt phagosome maturation in macrophage cell lines as well as to modulate and interfere with T lymphocyte immunological functions. Gastric mucosa as well as the H. pylori-infected epithelial cell line AGS strongly express IL-8 receptor A and B, which might contribute to the augmentation of the inflammatory response. Accumulating evidence implicates genetic variation in the inflammatory response to H. pylori in the etiology of the increased risk of gastric cancer after H. pylori infection. The chronic imbalance between apoptosis and cell proliferation is the first step of gastric carcinogenesis. In this regard, it was demonstrated that coexpression of two H. pylori proteins, CagA and HspB, in AGS cells, caused an increase in E2F transcription factor, cyclin D3, and phosphorylated retinoblastoma protein. Taken together, we now have a better understanding of the role of different virulence factors of H. pylori. There is still a lot to be learned, but the promising discoveries summarized here, demonstrate that the investigation of the bacterial survival strategies will give novel insights into pathogenesis and disease development. PMID- 15347302 TI - Inflammation, immunity and vaccines for Helicobacter pylori. AB - Helicobacter pylori causes chronic gastritis in the human stomach, yet only a minority of infected individuals develop peptic ulcer disease, atrophic gastritis, or gastric malignancies. The severity, progression, and consequences of H. pylori infection have been shown to depend on the host genetic background, and in particular on gene polymorphisms affecting the host immune response. Numerous studies published last year brought new information on the mechanisms by which the host genetic make-up modifies the inflammatory and immune responses to H. pylori and the induction of tissue damage secondary to the infection. Novel insights on the regulatory role of H. pylori on the adaptive T-cell response and on its consequences for the persistence of the infection and for the development of vaccines are discussed. PMID- 15347303 TI - Helicobacter pylori and nonmalignant diseases. AB - This paper reviews the new literature from the past year on the association between colonization with Helicobacter pylori and non-malignant disease of the upper gastrointestinal tract. This issue has, in the past year, remained a topic of wide research interest yielding many important new data. These data show that H. pylori eradication is the most effective therapy for peptic ulcer disease, but that a considerable proportion of ulcer patients remain to have dyspeptic symptoms. The discussion on the interaction between H. pylori and NSAID use in the etiology of ulcer disease has not yet been settled. Several studies, both from Asia and Europe, now reported that H. pylori eradication has a minimal effect on the primary prevention of ulcer disease in NSAID users, but eradication appears of relevance for the secondary prevention of ulcer disease in addition to proton pump inhibitor maintenance therapy. Various studies brought further support for the hypothesis that H. pylori eradication is of some benefit for patients with non-ulcer dyspepsia, although the effects are limited. The prevalence of H. pylori is lower among GERD patients than among controls, but H. pylori eradication has not been consistently shown to increase the risk for the newly development of GERD in an individual subject undergoing H. pylori eradication. The discussion on H. pylori and GERD should not preclude us from treating H. pylori-infected patients for accepted clinical indications. In patients using proton pump inhibitors for GERD, H. pylori eradication leads to a resolution of their corpus-predominant pangastritis, without impairing the efficacy of PPI therapy. PMID- 15347304 TI - Helicobacter pylori and gastric malignancies. AB - Clinical outcomes of Helicobacter pylori infection are diverse and caused by the variability of H. pylori virulence factors, host susceptibility, environmental factors and their interactions. Prospective epidemiological studies have clearly shown the relationship between H. pylori infection and gastric cancer. In addition, studies in animal models such as Mongolian gerbils with or without the addition of low-dose chemical carcinogens demonstrated that H. pylori infection can develop gastric carcinoma. Experimental studies have shown that virulence factors of H. pylori interact with gastric epithelial cell signaling related to carcinogenesis. While the effect of H. pylori eradication on prevention of such tumors in chronic H. pylori carriers is still controversial, progress has been made in therapy especially of gastric lymphoma with well tolerated chemotherapeutic regimen. PMID- 15347305 TI - Therapy of Helicobacter pylori. AB - This represents an overview of the main data published over the last year on the therapy of Helicobacter pylori. The problem of increasing failure of H. pylori eradication has been the main focus, with increasing resistance and poor patient compliance being the main culprits. Simple regimens are necessary to improve patient compliance. New antibiotics and novel agents are appraised with mixed results. PMID- 15347306 TI - Helicobacter pylori infection in pediatrics. AB - The proposed route of Helicobacter pylori transmission appears to be fecal-oral, oral-oral and gastro-oral, yet, a number of studies challenged these hypotheses in 2003. The use of the stool antigen test and[13]-C urea breath testing were the tests of choice for diagnosis and 'test for cure' of H. pylori in Europe in 2003 but have not yet become accepted standard of care in North America. Pediatric H. pylori consensus guidelines are not yet revised; upper endoscopy and biopsy remain the gold standard for diagnosis of pediatric H. pylori infection. In addition to stronger evidence supporting the role of host influences of H. pylori associated gastric cancer risk, compelling evidence was provided for the role of H. pylori in iron deficiency anemia of childhood. Antibiotic resistance remains a problem in conferring treatment failure and 2003 studies indicate that macrolide resistance is higher in children than in adults. Treatment with proton pump inhibitor-based triple therapy for 10-14 days remains the mainstay for eradication of H. pylori in childhood. Finally, multinational studies are needed to develop screening guidelines for childhood infection to avoid long-term severe gastroduodenal disease sequelae. PMID- 15347307 TI - Helicobacter pylori and extragastric diseases--other Helicobacters. AB - Reports on Helicobacter pylori and extragastric diseases have almost doubled this year compared with last year, bearing witness to the persistent scientific interest in this branch of Helicobacter-related pathology. Data belong increasingly to the area of vascular medicine, as well as hematology, dermatology, pediatrics and other fields. Unfortunately, these studies show overall controversial results, due to the impact of several confounding factors, and to the difficulty of recruiting homogeneous patient populations. Furthermore, many studies continue to be conducted on Helicobacter species other than H. pylori, focusing on animal models of gastroenterological illnesses which may retain strong similarities with human diseases. In this paper, taxonomy, detection and characterisation of Helicobacter spp. will be reviewed, together with the most important data issued this year on other Helicobacters and animal models. PMID- 15347308 TI - Helicobacter pylori public health implications. AB - Population Helicobacter pylori screening and treatment has the potential to dramatically reduce global gastric cancer mortality. There is overwhelming evidence that the infection is a major cause of distal gastric adenocarcinoma. There is also randomized controlled trial evidence that H. pylori eradication reverses or ameliorates histological changes in the gastric mucosa that are important in carcinogenesis. Preliminary randomized controlled trial data suggest that screening and treatment may reduce the risk of gastric cancer although the number of cancer cases was small. Population H. pylori screening and treatment will also reduce mortality from peptic ulcer complications and reduce the burden of dyspepsia in the community. The reduction in health service dyspepsia costs means that this could be the first programme to pay for itself. From a scientific perspective, we still have insufficient evidence to conclude the benefits of population H. pylori screening are greater than the possible harms and we need more randomized controlled trial data. From a public health perspective however, sometimes screening programmes are developed with imperfect information. The medical community should be consistent and if we are instituting other population screening programmes without randomized controlled trial evidence then H. pylori testing and treatment should also be considered. PMID- 15347309 TI - Bleeding profiles of anticoagulants, including the novel oral direct thrombin inhibitor ximelagatran: definitions, incidence and management. AB - Ximelagatran is a novel oral direct thrombin inhibitor (oral DTI) that is currently in advanced clinical development for the prevention and treatment of thromboembolic events in a wide range of patient populations and indications. The clinical development of novel anticoagulant therapies requires that treatments be assessed according to both their clinical benefit (reduction of risk of thromboembolic events) and safety profile (primarily bleeding). Definition and assessment of bleeding severity is thus an important factor in clinical trial design. Lack of consistency in bleeding definitions used in different clinical trial programmes makes comparison of bleeding event data difficult. Standard bleeding definitions would be required to make fair comparisons between clinical trials possible. The definitions of bleeding events used in clinical trials of ximelagatran are broadly consistent with those used in many other major trials. Results of phase II and III trials comparing ximelagatran with currently available anticoagulant therapies demonstrate that ximelagatran can be used with fixed dosing with no coagulation monitoring, dose titration, or dose adjustment, without compromising efficacy or safety. The incidences of major bleeding events in clinical trials of ximelagatran have been low and similar to those with other anticoagulant drugs. Adequate treatment in case of emergency situations such as serious bleeding should include cessation of treatment and maintenance of adequate diuresis. PMID- 15347310 TI - Assessment of the response to imatinib in chronic myeloid leukemia patients- comparison between the FISH, multiplex and RT-PCR methods. AB - OBJECTIVE: The objective of this study was to evaluate the kinetics of molecular response in chronic myeloid leukemia (CML) patients treated with imatinib and to compare between the fluorescent in situ hybridization (FISH), multiplex and real time quantitative RT-PCR (RQ-PCR) methods with this respect. METHODS: Molecular follow-up was carried out on 24 CML patients treated with imatinib. FISH analysis was performed according to the standard protocol. For RT-PCR the multiplex and RQ PCR methods were used. RESULTS: Sixty-three percent and 52% of the patients achieved complete remission according to FISH and multiplex RT-PCR analyses, respectively. Seventy-five percent of the patients achieved remission within the first year of treatment. In 83% of the cases the FISH and RT-PCR results were concordant. RQ-PCR analysis was carried out on 32 of the 41 samples negative by multiplex RT-PCR but only nine were negative. All samples with a BCR-ABL/ABL ratio below 2% were also negative by FISH. There was an excellent correlation between the RQ-PCR and the FISH tests. CONCLUSIONS: Molecular remission according to FISH and multiplex RT-PCR can be achieved by imatinib within 1 yr of therapy. There is a good correlation between the FISH, multiplex and RQ-PCR results in terms of the kinetics of disappearance of the BCR-ABL transcript and the predictability of each method for the other. Although RQ-PCR is the most sensitive method for molecular follow-up, FISH and multiplex RT-PCR can be used as complementary tools, at least during the early period of treatment. PMID- 15347312 TI - Anti-CD20 monoclonal antibody therapy in relapsed MALT lymphoma of the conjunctiva. AB - Low-grade non-Hodgkin's lymphomas of the conjunctiva may be cured by radiotherapy, but complications are frequent and relapses may occur. Other treatment modalities including resection, cryotherapy, injection of interferon alpha or systemic chemotherapy have been used with varying success. We treated two patients with relapsed extranodal marginal zone lymphoma (ENMZL) of mucosa associated lymphoid tissue (MALT) of the conjunctiva with the anti-CD20 monoclonal antibody rituximab (375 mg/m2 intravenously once weekly for 4 wk) which has previously been shown to be effective in a variety of other B-cell non Hodgkin's lymphomas. Treatment was well tolerated and resulted in one partial and one complete remission. With a follow-up of 32 or 30 months, respectively, further recurrences have not been observed. Rituximab is a highly effective and well-tolerated treatment of conjunctival MALT lymphoma, which may not only be of value in relapse, but also in cases of contraindication to radiotherapy. PMID- 15347311 TI - Significant thrombocytopenia associated with the addition of rituximab to a combination of fludarabine and cyclophosphamide in the treatment of relapsed follicular lymphoma. AB - Fludarabine in combination with cyclophosphamide is an effective treatment for newly diagnosed as well as relapsed follicular lymphoma. The anti-CD20 antibody rituximab has been employed successfully for the same indications. No such data were available on a combined use of these agents. Therefore, we conducted a phase II study to evaluate the safety and efficacy of a combination of rituximab (375 mg/m2), fludarabine (4 x 25 mg/m2) and cyclophosphamide (1 x 750 mg/m2), for the treatment of relapsed follicular lymphoma. An unexpected, severe hematologic toxicity with significant, prolonged thrombocytopenias WHO grade III/IV in 6 (35%) of 17 patients treated in total occurred, leading to early termination of the trial. Cytologic and serologic analyses point toward a direct toxic effect. Older patients (mean age 64.7 vs. 56.5 yr) were significantly (P = 0.02) more likely to suffer from this toxicity, whereas no other clinical or hematologic parameter differed statistically between the patients suffering from thrombocytopenia and those who did not. The addition of rituximab to fludarabine/cyclophosphamide employed at doses given above in relapsed follicular lymphoma may have led to this increase in thrombocytopenias. Therefore, caution should be exercised when combining these drugs for the treatment of patients with relapsed follicular lymphoma, especially when treating older patients. PMID- 15347313 TI - Relation between harvest success and outcome after autologous peripheral blood stem cell transplantation in multiple myeloma. AB - We studied the prognostic influence of pretransplant characteristics on response and survival in 104 consecutive patients with multiple myeloma receiving uniform pretransplant treatment consisting of VAD regimen, stem cell mobilisation, harvesting, and conditioning with melphalan 200 mg/m2. At the time of peripheral blood stem cell transplantation (PBSCT), 11% of patients were in complete remission (CR), and 63% in partial remission (PR). We evaluated the influence of age, sex, pretransplant response, number of harvested CD34+ cells, number of infused CD34+ cells, splitting part of the harvest for succeeding transplants on overall (OS) and progression-free survival (PFS) times. Following PBSCT, 31% of the patients were in CR and 57% in PR. Median OS and PFS from transplantation were 67 and 36 months, respectively. Transplant-related mortality was 0%. The number of harvested CD34+ cells was the only variable that was prognostic for OS in univariate and multivariate analyses. None of the variables was prognostic for PFS, although pretransplant response was nearly significant. The procedure of splitting and saving part of the harvest thus reducing the number of cells in the graft had no influence on outcome measured as OS or PFS. PMID- 15347314 TI - Pharmacokinetics of pegylated recombinant human megakaryocyte growth and development factor in healthy volunteers and patients with hematological disorders. AB - OBJECTIVES: The purpose of this study is to examine the pharmacokinetics of pegylated recombinant human megakaryocyte growth and development factor (PEG rHuMGDF) in healthy volunteers with normal hematopoiesis and patients with idiopathic thrombocytopenic purpura (ITP), acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and aplastic anemia (AA). METHODS: PEG-rHuMGDF was intravenously administered to healthy volunteers and patients with ITP, AML, MDS, and AA. The serum concentration of PEG-rHuMGDF was measured and the pharmacokinetics was investigated using non-linear mixed-effects modeling technique. RESULTS: The systemic clearance (CL) and volume of distribution at steady-state (Vss) consistently decreased in the healthy subjects, when the dose increased. In AML patients, CL and Vss decreased when the dose increased, but the change of CL was not statistically significant. In contrast, no significant dose dependency of these parameter estimates was observed in MDS patients. In AA patients there was no significant change in Vss but the CL of the higher dose groups was slightly smaller than that of the lower dose groups. Relatively smaller CL and Vss in ITP patients than those of healthy volunteers at the same dose were observed. CONCLUSIONS: This saturable pathway of CL may involve the receptor-mediated endocytosis and degradation by megakaryocyte lineage cells and platelets. The saturable distribution space can be also explained by the receptors on hematopoietic cells. The non-saturable distribution space corresponds to the value of plasma and interstitial fluid volume. PMID- 15347315 TI - Combined real time PCR and immunohistochemical evaluation of sperm protein 17 as a cancer-testis antigen. AB - We previously identified sperm protein 17 (Sp17) as a normal testicular protein aberrantly expressed in a proportion of multiple myeloma (MM). However, recent studies have generated controversies on the normal tissue expression of Sp17 and whether or not it is a suitable target for immunotherapy. In this study, we have used a combination of real time polymerase chain reaction and immunohistochemistry on a large panel of normal tissues. Although Sp17 transcripts could be detected in some normal tissues, the levels of expression were <2% of those in normal testis. In contrast, Sp17+ myeloma cells expressed 3 18% of normal testis levels of Sp17 transcript. Immunohistochemistry using two Sp17 murine monoclonal antibodies, each directed at a non-overlapping B-cell epitope, showed Sp17 protein to be expressed only in testis and not any other normal tissues. Specificity of binding of the antibodies to testis was also confirmed in competitive binding assays. Our results therefore further suggest Sp17 as a cancer-testis antigen in MM and support its suitability as a target for immunotherapy. PMID- 15347316 TI - Development of packaging cell lines for generation of adeno-associated virus vectors by lentiviral gene transfer of trans-complementary components. AB - Adeno-associated virus (AAV) vector system has several useful advantages with regard to in vitro and in vivo gene transfer. However, their usages have been limited by cumbersome and labor-intensive vector production in the traditional method. To overcome limitations in AAV production, in this report, we explored the possibility of generating AAV packaging cell line, 293T R/C.VA.E2A.E4. cells, by using lentivirus-mediated transduction of Rep/Cap gene of AAV-2, VA RNA, E2A, and E4 genes of Ad5 into 293T cells. In packaging cell lines, it is important that supply of the AAV vector can be stably performed for long time. We showed that the 293T R/C.VA.E2A.E4. cells have stably maintained the transduced components after more than 10 passages and yielded high-titer AAV vectors, and the titer of AAV vectors did not decline even if culture of the packaging cells was continued for long time. The Rep/Cap and E4 gene products caused no remarkable cytotoxicity. The 293T R/C.VA.E2A.E4. cells might be able to tolerate the Rep/Cap and E4 gene products, or have less copy numbers of the Rep/Cap and E4 genes than the traditional method. Moreover, we showed that the AAV vectors derived from 293T R/C.VA.E2A.E4. cells infected the primary human CD34+ haematopoietic progenitor cells with high efficiency (50-70%). In the 293T R/C.VA.E2A.E4. cells, the AAV vectors can be generated by the transfection of one AAV vector plasmid, and large-scale AAV production can be easily achieved. It is important that cumbersome, variable, and costly transfection is avoided. PMID- 15347317 TI - Flow-cytometric approach to the prompt laboratory diagnosis of TRALI: a case report. AB - OBJECTIVES: Transfusion-related acute lung injury (TRALI) is a rare but serious complication which can occur after transfusion of blood components. In this report we describe our flow-cytometry approach to the laboratory diagnosis of a case of TRALI in a recipient of fresh frozen plasma containing human leukocyte antigen (HLA) class II antibodies. METHODS: The post-transfusion reaction work-up included the direct and indirect Granulocyte Immunofluorescence Test (GIFT) on the recipient's neutrophils collected before and after the reaction and on the serum from the recipient and from all implicated donors; flow-cytometry bead based screening and identification assay for HLA class I and II antibodies in donor sera and flow cytometry cross-matching on T and B patient's lymphocytes. Finally, we investigated the reactivity of one donor serum, containing HLA class II antibodies, with the patient's neutrophils activated in vitro to induce expression of HLA class II. RESULTS: We found an increased level of IgG bound on patient's granulocytes collected after TRALI, in the absence of detectable granulocyte and HLA class I antibodies in the five implicated donors. One of them showed HLA-DR 1 and -DR 51 antibodies, which determined a positive cross-match with patient's B lymphocytes and in vitro activated granulocytes. Both HLA class II antigens were present in the recipient and absent in the donor. CONCLUSIONS: In some pathological conditions, HLA class II antibodies can react with activated granulocytes expressing HLA-DR antigens, and activate TRALI reaction. HLA class II antibodies screening and flow cytometry cross-matching techniques should be added to the current diagnostic algorithm of TRALI. PMID- 15347318 TI - A sustained response to low dose interferon-alpha in a case of refractory pure red cell aplasia. AB - Acquired pure red cell aplasia (PRCA) may be the result of a cellular or humoral autoimmune process. One proposed mechanism is the destruction of erythroid progenitors by self-reactive, cytotoxic T cells or natural killer (NK) cells. These cells normally express MHC class I receptors (KIR) which inhibit cytotoxicity when the target cell expresses the HLA class I antigen(s) they bind. Therefore, loss of these antigens on maturing erythroid progenitors may render them susceptible to destruction by the pathogenic cells. Interferon-alpha (INF alpha) increases HLA class I expression on hematopoietic precursor cells. Therefore, we initiated a trial of INF-alpha in a patient with refractory PRCA. Following treatment, he developed transfusion independence, and a sustained normal hematocrit. Analysis of bone marrow erythroid cells revealed an increase in expression of HLA class I molecules. INF-alpha should be used in a controlled trial in patients with PRCA to determine its activity and mechanism of action. PMID- 15347319 TI - Paroxysmal cold haemoglobinuria as a tardive complication of idiopathic myelofibrosis. AB - Paroxysmal cold haemoglobinuria (PCH) is an autoimmune haemolytic anaemia caused by the Donath-Landsteiner antibody. It is classically described in association with chronic syphilis or after acute viral infections. We describe the first case of PCH presented as a late manifestation of advanced myelofibrosis associated with antiphospholipid syndrome, that promptly responded to high dosage of prednisone. PMID- 15347320 TI - Expression of ceramide kinase in hematopoietic cells. PMID- 15347322 TI - Skin imaging: is it clinically useful? AB - Non-invasive skin imaging techniques have proliferated over the last decade. Whilst most have a research role, some are routinely used in dermatology clinics. Of these, the skin surface microscope (dermatoscope), a diagnostic aid for pigmented lesions, has had most clinical impact. Such devices, when linked to a videomicroscope for computer analysis, have been dubbed as 'mole scanners'. Mole scanners are increasingly available on a commercial basis even though computer diagnosis of pigmented lesions is currently no better than diagnosis by human experts. Meanwhile, other imaging techniques, such as high-resolution ultrasonography, spectroscopy and optical coherence tomography, may yet find a role in diagnosis and disease monitoring. PMID- 15347323 TI - Marie Unna hereditary hypotrichosis: report of a Chinese family and evidence for genetic heterogeneity. AB - Marie Unna hereditary hypotrichosis (MUHH) is a rare autosomal dominant disorder with progressive hair loss starting in early childhood and aggravating at puberty. Several studies have mapped the MUHH gene to chromosome 8p21. Here we report a Chinese MUHH family with variable phenotypes. All affected individuals have anomalies affecting both hair density and hair shafts. Major clinical characteristics, disease history and histological examination support the diagnosis of MUHH, but the features of scarring in this kindred are modest and none of the patients have vertex hair loss, which is in contrast with typical MUHH. We now report genotyping and linkage analysis using 11 polymorphic microsatellite markers spanning the MUHH locus at 8p. Two-point linkage analysis using these markers revealed significant exclusion of this locus (log of the odds scores < - 2) at Theta = 0 indicating that there is a range of clinical presentations in MUHH, and that more than one genetic locus is responsible for the disorder. PMID- 15347324 TI - The psychological impact of cutaneous leishmaniasis. AB - A psychiatric disorder would be associated with extensive, unsightly lesions on exposed body parts. Cutaneous leishmaniasis (CL) has long been endemic in Sanliurfa and is called 'beauty scar'. The aim of this study was to determine psychological impact of CL. Patients with active CL, with CL that had healed with scaring, and healthy controls were included in this case-control study. The Hospital Anxiety Depression Scale (HAD), Body Image Satisfaction Scale (BIS), and Dermatology Quality of Life Scale (DQL) assessments were performed to determine the psychological effect of CL. The patients with CL had significantly higher HAD anxiety and depression subscale scores than the control groups. Patients with CL have decreased body satisfaction and lower quality of life than those in the control group. It was found that CL patients with active lesions have the lowest quality of life score than other groups. CL lesions on exposed body parts such as the face and hands, active CL for more than 1 year, permanent scar formation, and social stigmatization cause anxiety, depressive symptoms, decreased body satisfaction and quality of life in CL patients. PMID- 15347325 TI - Cutaneous leukaemic infiltrations in a patient with previously undiagnosed myelodysplastic syndrome. AB - We report the rare case of a patient with leukaemia cutis first presenting only on the hand and fingers and then subsequently spreading over the trunk and face. The lesions heralded the transformation of a previously undiagnosed myelodysplastic syndrome type RAEB (refractory anaemia with blast excess) into frank myeloid leukaemia. The haematological disease was first detected by the dermatohistopathologist. This case underlines the need to look meticulously for skin changes and perform early skin biopsies in haematological patients, as the skin can reveal the first clinical signs of an otherwise not evident bone marrow disorder. Leukaemia cutis as the initial clinical presentation of a transforming myelodysplastic syndrome type RAEB into acute myeloid leukaemia has been reported only very rarely. PMID- 15347326 TI - Kissing naevus of the penis. AB - Divided or 'kissing' naevus was first described on the eyelids in 1908. Other types of divided naevus reported include naevus spilus on the eyelids, mast cell naevus and epidermal naevus in a divided form on the fingers. Six cases of kissing naevus of the penis appear in the literature. In this paper, we discuss a seventh case. PMID- 15347327 TI - Multiple congenital melanocytic naevi presenting with neurofibroma-like lesions complicated by malignant melanoma. AB - Giant congenital pigmented naevi and neurofibromatosis (NF-1) may rarely occur together. We report an unusual case where extensive congenital melanocytic naevi were associated with neurofibroma-like lesions that were clinically and histologically confused with neurofibromatosis. The development of malignant melanomas within the pigmented and pendulous lesions representing multiple congenital melanocytic naevi highlights the importance of an accurate diagnosis and a close follow-up of such patients. PMID- 15347328 TI - Hallermann-Streiff syndrome: a case review. AB - Hallermann-Streiff Syndrome is a rare genetic disorder that is characterized primarily by head and face abnormalities. Patients show bird-like facies, dental abnormalities, and hypotrichosis with various ophthalmic abnormalities. We report here a 26-year-old woman with Hallermann-Streiff Syndrome and review the literature. PMID- 15347329 TI - Spontaneous clinical improvement in HIV-associated follicular syndrome. AB - Since 1991 infrequent reports have described a distinctive triad of nodulocystic acne, striking follicular spines and an eruption resembling pityriasis rubra pilaris (PRP) in HIV-positive patients. It has been suggested that this may represent a subtype of PRP, or alternatively that it should be viewed as a unique HIV-associated follicular occlusion triad. Clinical manifestations may be severe, and in several cases have been ultimately fatal, with death occurring due to complications of cutaneous sepsis. We describe a case demonstrating severe conglobate acne, follicular keratotic spines and histologically confirmed PRP in association with HIV infection. Clinical features and treatment modalities of previously reported cases are reviewed. Despite refusing all topical and systemic treatment our patient showed spontaneous remission of skin signs after 2 years. PMID- 15347330 TI - Multiple dermatofibromas in a patient with systemic lupus erythematosus and Sjogren's syndrome. AB - Multiple dermatofibromas (DFs) are rare and have been thought to be associated with altered immunity. In this report, we describe a 27-year-old Japanese woman with systemic lupus erythematosus (SLE) and Sjogren's syndrome in whom eight nodules appeared over a period of 4 years. Histopathological findings were consistent with DF. SLE rather than Sjogren's syndrome seemed to have induced the multiple DFs in this patient. We also reviewed the reported cases with multiple DFs associated with SLE and/or Sjogren's syndrome. Review of the previous reports indicates that SLE is the most frequent autoimmune disorder associated with multiple DFs, and that both SLE and immunosuppressive treatments play a part in induction of multiple DFs. Therefore, if multiple DFs are present it is important that the status of the patient be evaluated from the standpoint of autoimmune diseases, particularly SLE, or immunosuppression. PMID- 15347331 TI - Ectodermal dysplasia showing clinical overlap between AEC, Rapp-Hodgkin and CHAND syndromes. AB - The ectodermal dysplasias represent a complex collection of congenital abnormalities of skin, hair, teeth, nail, and sweat gland development, many of which have overlapping clinical features. In this report, we describe a 7-year old girl, born to clinically normal parents, with ankyloblepharon, cleft lip/palate and hair abnormalities, features resembling the autosomal dominant disorder, ankyloblepharon-ectodermal dysplasia-clefting (AEC) syndrome, which results from mutations in the sterile-alpha motif domain of the gene encoding the transcription factor, p63. However, direct sequencing of the p63 gene in this individual did not reveal any pathogenic sequence variants. Moreover, two of her paternal cousins were discovered to have similar congenital ectodermal anomalies, raising the alternative possibility of an autosomal recessive pattern of inheritance. Furthermore, all affected individuals lacked a history of erosive scalp dermatitis that is usually characteristic of AEC syndrome. Instead, the scalp hair was coarse and wiry. In addition, another atypical feature, hypohidrosis, was present. Collectively, the clinical features also resembled Rapp-Hodgkin syndrome, Bowen-Armstrong syndrome and CHAND syndrome, but did not appear to fit neatly with any one particular disorder. This case highlights the difficulties in trying to classify the ectodermal dysplasia syndromes on clinical features alone. PMID- 15347332 TI - Multiple scrotal epidermolytic acanthomas; secondary to trauma? AB - Epidermolytic hyperkeratosis (EH) is an abnormality of epidermal maturation, most commonly due to mutations in keratins 1 and 10, which may be a congenital or an acquired defect. The term epidermolytic acanthoma was applied to a solitary discrete epidermal proliferation characterized by EH. Subsequently there have been several reports of disseminated epidermolytic acanthomas. We report a rare case of multiple epidermolytic acanthomas localized to the scrotum. With the aetiology of epidermolytic acanthoma unknown, trauma has been postulated as a possible cause. Our patient repetitively scratched his scrotum for 5 years and we believe that this action triggered his multiple scrotal epidermolytic acanthomas. PMID- 15347333 TI - Post-operative delayed bleeding from the superficial temporal artery. AB - We present three patients who experienced late, severe bleeding from the superficial temporal artery (STA) following uncomplicated removal of basal cell carcinoma (BCC) from the temple. This is a common site for BCC and the risk of damage to the temporal branch of the facial nerve and the STA are well known. However, delayed bleeding as described in these individuals is potentially dangerous but poorly reported. PMID- 15347334 TI - Epidermal naevi treated with pulsed erbium:YAG laser. AB - Epidermal naevi are common and can cause marked cosmetic disability. Ablative laser treatment may be successful in their removal, but may result in significant scarring. Surprisingly the erbium:YAG laser is rarely reported for this indication even though it produces minimal tissue damage. We report our experience using this laser to treat six patients with epidermal naevi. Patients (five female, one male; aged 4-41 years) underwent treatment with pulsed 2940 nm erbium:YAG laser at 0.4-0.45 J/cm(2), 2 mm spot size at 4 pulses/s. All six patients had excellent cosmetic results at follow up ranging from 6 to 60 months. The favourable results were dependent on selection of cases with superficial or small, discrete lesions which could be ablated accurately. The erbium:YAG laser is therefore an effective treatment for relatively nonverrucous or papular epidermal naevi. PMID- 15347335 TI - A case of monocytic leukaemia cutis in a patient with myelodysplastic syndrome transforming to acute myeloid leukaemia. AB - A 49-year-old woman presented with a 1-year-history of a widespread eruption which proved to be due to leukaemia cutis. Subsequently, she developed pancytopaenia and a bone marrow biopsy revealed refractory anaemia with excess blasts in transformation (RAEB-T) with a high monoblastic component. This transformed into acute myeloid leukaemia. Leukaemia cutis in this context is well described but in this patient it became manifest 1 year prior to referral to the dermatologist. When occurring with a myelodysplastic syndrome, leukaemia cutis often heralds malignant transformation to acute myeloid leukaemia. Prompt diagnosis in this situation may identify a group of high-risk patients with myelodysplastic syndrome for whom chemotherapy and allogenic bone marrow transplantation, rather than the more conventional approach of supportive treatment, could be a more appropriate management strategy. PMID- 15347336 TI - Characterization of IgG autoantibodies to extracellular matrix protein 1 in lichen sclerosus. AB - Although the precise aetiology of lichen sclerosus is unknown, evidence for an autoimmune basis to the disorder is emerging. Indeed, circulating IgG autoantibodies to the glycoprotein extracellular matrix protein 1 (ECM1) have been demonstrated in the sera of about 75% of affected individuals. To assess this humoral immune response further, immunoblotting was performed using bacterial recombinant proteins spanning different domains of the ECM1 protein. The aim was to identify autoantibody-reactive sites recognized by 90 lichen sclerosus sera. The subclass distribution of anti-ECM1 IgG autoantibodies was also determined in 54 lichen sclerosus sera. Immunoblotting showed that the IgG autoantibodies from lichen sclerosus patients recognize multiple antigenic reactive sites on the ECM1 protein within both the amino terminus (50/90, 55.6%) and the protein loop cysteine-rich repeat domains (54/90, 60%), although few sera (7/90, 7.8%) had antibodies to the carboxyl terminus of ECM1. IgG subclass analysis revealed that the anti-ECM1 autoantibodies belong predominantly to the IgG(2) subclass (48/54, 88.9%), either IgG(2) alone (28/54, 51.9%) or in combination with one or more other IgG subclasses. No correlation was found between the site(s) of the ECM1 epitopes or the anti-ECM1 IgG profile and any specific clinical parameters. Nevertheless, characterization of anti-ECM1 antibodies does provide further insight into humoral immune responses and understanding disease mechanisms in lichen sclerosus. PMID- 15347337 TI - Topical imidazoquinoline therapy of cutaneous squamous cell carcinoma polarizes lymphoid and monocyte/macrophage populations to a Th1 and M1 cytokine pattern. AB - Imidazoquinolines are topical immune response modifiers. Imiquimod (IMI), the first imidazoquinoline, is approved for the treatment of genital human papillomavirus disease and has shown success as a therapeutic agent for cutaneous premalignant and malignant tumours. To define the pattern of polarization of the local immune response to invasive cutaneous squamous cell carcinoma (SCC) we pretreated 10 SCCs that were > 3 cm in diameter for 2 weeks with IMI. The tumours were treated on Monday, Wednesday and Friday and excised the next Monday. A battery of immunohistochemical markers was used to define the mononuclear cell populations in the diagnostic, and the excisional biopsy specimens. The total inflammatory infiltrate was increased after IMI therapy: the greatest increase was in the CD8 T cells with a marked relative decrease in the CD68 monocytic/macrophages; the majority of the CD8 T cells showed expression of cytotoxic granules, T cell-restricted intracellular antigen (TIA) and granzyme B. The relative numbers of monocytes/macrophages were decreased after therapy with IMI with a decrease in CD68+, CD23+, and CD14+ cells and an increase in MAC-397+, and factor XIIIa+ cells. The epidermal dendritic cells showed a > 75% decrease in CD1a+ cells. The immunohistochemical marker profile after IMI therapy is consistent with that induced by a Th1 and M1 cytokine polarization pattern. This cytokine pattern is known to be more effective in defence against tumours as well as viral infections. PMID- 15347338 TI - Deleterious mutations in SPINK5 in a patient with congenital ichthyosiform erythroderma: molecular testing as a helpful diagnostic tool for Netherton syndrome. AB - The congenital erythrodermas represent a heterogeneous group of inherited and acquired disorders often accompanied by systemic infections, impaired epidermal barrier function and concomitant life-threatening fluid and electrolyte imbalance. In the present report, we describe a patient who was considered to have congenital ichthyosiform erythroderma for 26 years until molecular testing led to the correct diagnosis of Netherton syndrome. PMID- 15347339 TI - Open study of the efficacy and mechanism of action of topical imiquimod in basal cell carcinoma. AB - Imiquimod is an immune-response modifier that has been shown to be effective in the treatment of superficial and nodular basal cell carcinoma (BCC). The objective of this open-label study was to investigate the effectiveness of imiquimod 5% cream in superficial, nodular, and infiltrative BCC. Fifty-five Caucasian patients with primary BCC measuring 8 mm or more in diameter with a superficial, nodular, or infiltrative histological pattern were included in the study. Four groups of BCC (A, B, C, and D) and two dosing regimens were studied: 35 BCCs (groups A, B, and C) were treated with imiquimod three times weekly and 20 BCCs (group D) were treated with imiquimod five times weekly. Histological samples were obtained before treatment, during treatment (on day 22 in group A, day 15 in group B, and day 8 in groups C and D), and 6 weeks after treatment. All patients were followed-up for a minimum of 2 years. In the biopsy specimens obtained, the expression of Bcl-2, p53, and Ki-67, apoptotic index (Tunel technique), and the number of CD3+, CD8+, CD20+, CD56+, CD68+, granzyme B+, and S 100+ cells in the peritumoural inflammatory infiltrate, were determined and quantified. Of the 55 BCCs treated with imiquimod 41 (74%) were in complete remission after 2 years of follow-up. These comprised 4/4 superficial BCCs, 7/8 (88%) nodular BCCs, and 30/43 (70%) infiltrative BCCs. Multi-variate analysis demonstrated that baseline tumour size was the most powerful independent prognostic variable (P < 0.05). Treatment with imiquimod increased the apoptotic index (P < 0.05), reduced Bcl-2 expression (P < 0.05), and increased the number of CD3+, CD8+, CD20+, CD68+, granzyme B+, and S-100+ cells in the inflammatory infiltrate of the BCC (P < 0.05). In conclusion, imiquimod induced an antitumour immune response mediated by lymphocytes and macrophages, reduced Bcl-2 expression and increased the apoptotic index of BCC, and was clinically effective in 74% of BCCs after a 2-year follow-up period. PMID- 15347340 TI - Effects of bepotastine, cetirizine, fexofenadine, and olopatadine on histamine induced wheal-and flare-response, sedation, and psychomotor performance. AB - Although many antihistamines are now in clinical use, few studies directly compare their pharmacodynamic and sedative activities in humans in vivo. We designed a double-blind, placebo-controlled, crossover study to compare the inhibitory effects of bepotastine, cetirizine, fexofenadine, and olopatadine on histamine-induced flare-and-wheal response. Systemic sedative effects and impaired psychomotor activities by these drugs were also evaluated. Bepotastine (10 mg twice a day), cetirizine (10 mg once a day), fexofenadine (60 mg twice a day), and olopatadine (5 mg twice a day) or placebo was given in a double-blind manner to seven healthy volunteers before histamine challenge by iontophoresis. At 0, 1, 2, 4, 8, 12, and 24 h following the oral administration of these drugs, histamine iontophoresis-induced wheal-and-flare response was measured. Sedative effects by the drugs were also evaluated by a visual analogue scale for subjective sedation, and by word processor test for psychomotor activity. Each volunteer was tested with all of the drugs (including placebo), administered in a random order with a washout period of at least 1 week. Histamine iontophoresis induced marked wheal-and-flare response in all participants. Bepotastine, cetirizine, fexofenadine, and olopatadine yielded significant reduction of histamine-induced wheal-and-flare response compared to placebo (P < 0.01). Among the drugs, olopatadine and cetirizine suppressed most markedly and persistently histamine-induced wheal-and-flare response, while bepotastine and fexofenadine produced a significant, but less persistent suppression. Olopatadine, fexofenadine, and cetirizine showed a significant systemic sedative effect in this order with bepotastine showing the least sedative effect. Moreover, olopatadine affected psychomotor performance most markedly, which was followed by fexofenadine and cetirizine. These results indicate that bepotastine, cetirizine, fexofenadine, and olopatadine inhibit histamine-induced wheal-and-flare response of humans in vivo and induce a variable systemic sedative effect and impaired psychomotor activity. Although olopatadine and cetirizine showed the strongest and most persistent suppression of histamine-induced wheal-and-flare response, olopatadine showed a considerable sedative effect with impaired psychomotor performance. PMID- 15347341 TI - A novel mutation of the DSRAD gene in a Chinese family with dyschromatosis symmetrica hereditaria. AB - Dyschromatosis symmetrica hereditaria (DSH) is a pigmentary genodermatosis of autosomal dominant inheritance characterized by a mixture of hyperpigmented and hypopigmented macules distributed on the dorsal aspects of the hands and feet. It is caused by mutations of the RNA-specific adenosine deaminase gene. We report the identification of a Chinese family with a three-generation pedigree of DSH, in whom a novel tyrosine substitution mutation in DSRAD was demonstrated: a heterozygous nucleotide A-->G transition at position 2879 in exon 10 of the DSRAD gene was detected. PMID- 15347342 TI - A novel 7-bp deletion mutation in a Taiwanese family with X-linked hypohidrotic ectodermal dysplasia. AB - Hypohidrotic ectodermal dysplasia (HED) is found worldwide with an estimated incidence of 1 per 100,000 births. X-linked hypohidrotic ectodermal dysplasia (XLHED, OMIM 305100) is the most common form of the ectodermal dysplasias (ED), a rare group of hereditary diseases characterized by abnormal development of eccrine sweat glands, hair, and teeth. Heterozygous carriers of XLHED often manifest minor or moderate degrees of hypotrichosis, hypodontia, and hypohidrosis. ED1, the gene for XLHED encodes ectodysplasin A, which is a new member of the tumour necrosis factor family. The majority of mutations in XLHED are missense mutations, but one-fifth are insertion/deletions. Here we report a novel 7-bp deletion mutation (nt1242-1248) in exon 9 of the ED1 gene that results in a frameshift and premature stop codon (PTC + 38 amino acids). Mutation analysis in families with XLHED allows for genetic counselling, prenatal diagnosis and confirmation of carrier status. PMID- 15347343 TI - A new mutation in the linker 12 domain of keratin 5 in a Chinese family with Weber-Cockayne epidermolysis bullosa simplex. AB - A previously undescribed missense mutation was detected in the L12 domain of keratin 5 (K5) in a Chinese family with Weber-Cockayne epidermolysis bullosa simplex. Direct sequencing of the PCR products identified a single base substitution (983A-->G) that changes the aspartic acid residue at codon 328 to glycine in all affected family members, while no mutation was observed either in the healthy individual or 50 unrelated control samples. Asp328 of K5 is remarkably conserved among all type II keratins. D328G is the fourth mutation found to affect this residue in K5-related epidermolysis bullosa simplex, indicating the importance of Asp328 for K5 structure and the dramatic effect that fine changes can have on keratin intermediate filament integrity. PMID- 15347344 TI - NBCCS secondary to an interstitial chromosome 9q deletion. AB - Naevoid basal cell carcinoma syndrome (NBCCS) or Gorlin syndrome is a rare autosomal dominant cancer disorder. The Gorlin's gene, Patched 1 (PTCH1), maps to Chromosome 9q. Germline mutations of PTCH1 occur in patients with NBCCS. The subsequent loss of the remaining allele results in cancer formation. We present a patient with NBCCS and additional phenotypic features including severe developmental delay, short stature and hypotelorism who was found to have an interstitial chromosome 9q deletion. The NBCCS phenotype in our patient occurred as a result of PTCH1 deletion in contrast with an inherited mutation of this gene. PMID- 15347345 TI - Nail varnish is a potential allergen in nickel allergic subjects. PMID- 15347346 TI - Cutaneous leishmaniasis in a Kosovan child treated with oral fluconazole. PMID- 15347347 TI - Sarcoidosis presenting as tattoo changes in a patient undergoing treatment with interferon-alpha and ribavirin. PMID- 15347348 TI - Red pseudochromhidrosis of the neck. PMID- 15347349 TI - A case of psoriasis vulgaris with peripheral blood eosinophilia, parallelling the psoriasis area and severity index (PASI) score. PMID- 15347350 TI - Seronegative arthritis associated with the follicular occlusion triad. PMID- 15347351 TI - Scar sarcoidosis following tattooing of the lips treated with mepacrine. PMID- 15347352 TI - Transient eruptive seborrhoeic keratoses associated with erythrodermic pityriasis rubra pilaris. PMID- 15347353 TI - Cutaneous larva migrans acquired in Britain. PMID- 15347354 TI - Segmental neurofibromatosis on the face. PMID- 15347355 TI - Chylothorax after treatment with flash-lamp pulsed dye laser. PMID- 15347356 TI - Lack of efficacy and tolerability of topical PDT for psoriasis in comparison with narrowband UVB phototherapy. PMID- 15347357 TI - Trends in the presentation of cutaneous malignant melanoma over three decades at King's College Hospital, London. AB - The aim of this study was to examine trends in the presentation of cutaneous malignant melanoma at King's College Hospital (KCH) over the last three decades (1970-2000). KCH was one of seven centres that participated in the 1987 Cancer Research Campaign (CRC) publicity campaign aimed at promoting earlier self recognition of melanoma. Data included patient age at presentation, sex, tumour site, Breslow thickness and histological subtype. The late 1980s saw a threefold increase in the annual number of melanomas and an eightfold increase in thin melanomas compared to the 1970s. The increase occurred in both sexes and was particularly marked after the CRC campaign but numbers had already begun to increase prior to this. The increase has predominantly been thin (Breslow < 1.5 mm) tumours of the superficial spreading variety with a resultant fall in mean Breslow thickness. There has been a decline in the annual number of melanomas since the peak in 1992 which is not explained by increased proportion of in situ tumours. The CRC campaign may have contributed to the documented increase in thin tumours but this trend had begun prior to 1987 suggesting factors other than public awareness and earlier presentation are important. It is encouraging that the number of melanomas has declined over the last 5 years at KCH but it is yet to be seen whether this reflects a real decrease in the incidence of melanoma. PMID- 15347358 TI - Thyroid feeding: a forgotten treatment for psoriasis. PMID- 15347359 TI - Hand graft or hard luck. PMID- 15347360 TI - Competency assessment of dermatology trainees in the UK. AB - Postgraduate higher specialist training of dermatology in the UK has become more structured over the past 8 years. Increased awareness of the need to police our profession has impelled the introduction of objective competency based assessment. Competency assessment will take three forms: mini clinical evaluation exercise, directly observed procedural skills and 360-degree assessment. These components of assessment will direct much needed rigour into the training system and may be the first step on the road to a formal exit examination in dermatology. PMID- 15347361 TI - Learning the cost of dermatitis artefacta. PMID- 15347363 TI - Meeting the information needs of patients with allergic disorders: partnership is the key. PMID- 15347364 TI - Plant non-specific lipid transfer proteins as food and pollen allergens. AB - Several members of the plant non-specific lipid transfer protein (LTP) family have been identified as relevant allergens in foods and pollens. These allergens are highly resistant to both heat treatment and proteolytic digestion. These characteristics have been related with the induction of severe systemic reactions in many patients, and with the possibility of being primary sensitizers by the oral route. A specific geographical distribution pattern of sensitization to LTP allergens has been uncovered. This allergen family is particularly important in the Mediterranean area, but shows a very limited incidence in Central and Northern Europe. The potential role in the plant, as well as the biochemical and allergenic properties of the LTP family, are reviewed here. PMID- 15347365 TI - Anti-inflammatory properties of desloratadine. AB - BACKGROUND: Allergic rhinitis (AR) is associated with robust infiltration of immune cells and mediators that may contribute to clinical manifestations of the disease. OBJECTIVE: To review the complex immune effector mechanisms involved in the allergic response and discuss their effects on the pathophysiological and clinical manifestations of AR. Desloratadine, a novel antihistamine, was used as a probe with the goal of attaining a better understanding of the inflammatory processes underlying the allergic response. METHODS: Data were obtained from abstracts and peer-reviewed journals. The pathophysiology of the allergic response has been extensively studied. This paper presents only data from studies that used desloratadine at physiologically relevant concentrations. RESULTS: Key mediators involved in the allergic response and in pathophysiological and clinical manifestations of the immune response were reviewed. Desloratadine was used as a probe to further elucidate the mechanisms involved during an allergic response. CONCLUSIONS: Some have proposed a link between the pathophysiology of AR and the clinical manifestation of symptoms. Desloratadine, a new-generation antihistamine, has demonstrated anti-inflammatory effects in vitro; indeed, desloratadine is capable of intervening at various points in the immune cascade. Although in vitro results do not necessarily correlate with clinical efficacy, the anti-inflammatory properties of desloratadine may contribute to its efficacy in patients with AR, allergy-induced asthma, and other related allergic conditions. Antihistamines that modulate in the immune system at various stages may optimize treatment of allergic disease. PMID- 15347366 TI - Mode of delivery is not associated with asthma or atopy in childhood. AB - BACKGROUND: Caesarean-section delivery has been associated with the subsequent development of atopy and wheezing in childhood. OBJECTIVE: To examine the association between mode of delivery (vaginal vs. caesarean section) and development of atopy, asthma and wheezing disorders in a population-based cohort of children. METHODS: The Avon Longitudinal Study of Parents and Children is a longitudinal birth cohort of children born 1 April 1991 to 31 December 1992. Mode of delivery was categorized as vaginal (including forceps and ventouse extractions) or caesarean section (elective and emergency). Primary outcomes were parental report of asthma or wheezing between 69 and 81 months of age, physician diagnosed asthma (PDA) at 91 months of age and atopy at 7 years by skin prick testing. Possible confounding factors were considered in a multivariable logistic regression model. RESULTS: Total livebirths were 14,062, from which were selected 12 367 born to mothers resident in a defined area and delivered in one of two major obstetric hospitals. Of these infants, 10,980 (88.8%) were delivered vaginally and 1387 (11.2%) by caesarean section. Outcome data were available for 7495 (61%) subjects (asthma 69-81 months); 7389 (60%) (wheeze 69-81 months); 7196 (58%) (PDA 91 months) and 5916 (48%) (atopy 7 years). Adjusted odds ratios [95%confidence interval] for caesarean section compared with vaginal delivery were not statistically significant for any outcome we considered: asthma 69-81 months 1.16 [0.9, 1.5]; wheeze 69-81 months 0.95 [0.7, 1.3]; PDA 1.14 [0.9, 1.4]; atopy 1.04 [0.8, 1.3]. CONCLUSION: Delivery by caesarean section was not associated with the subsequent development of asthma, wheezing or atopy in later childhood in this population. PMID- 15347367 TI - Altered beta2-adrenergic regulation of T cell activity after allergen challenge in asthma. AB - BACKGROUND: Airway inflammation in asthma is orchestrated by recruitment of T helper (Th)2 lymphocytes to the lung and subsequent production of Th2-like cytokines upon allergen challenge. OBJECTIVE: To examine whether allergen-induced dysfunction of the beta2-adrenergic receptor (beta2-AR) contributes to the enhanced T(h2) cell activity in asthma. METHODS: Beta2-adrenergic regulation of cytokine mRNA expression was studied in alpha-CD3/alpha-CD28-activated peripheral blood lymphocytes from seven asthma patients before and 6 h after allergen challenge, in conjunction with the effects of beta2-agonist fenoterol on T cell chemotaxis and signalling pathways. RESULTS: A complete loss of beta2-AR control over expression of the Th2 cytokines IL-4, IL-5 and IL-13, but not of the Th1 cytokine IFN-gamma, was observed after allergen challenge. Furthermore, we found impaired beta2-AR regulation of T cell migration as well as signal transduction pathways, i.e. the phosphorylation of cyclic adenosine monophosphate-responsive element binding protein and the inhibition of the mitogen-activated protein kinase pathway. The loss of beta2-AR control was associated with increased beta adrenergic receptor kinase expression, which might be involved in beta2-AR desensitization. In addition, we demonstrate for the first time that T cells exposed to the chemokine thymus and activation-regulated chemokine show hyporesponsiveness to fenoterol. CONCLUSION: Our results suggest that allergen induced loss of beta2-AR control, possibly mediated by chemokine release, plays an important role in enhanced Th2-like activity in asthma. PMID- 15347368 TI - Defective suppression of Th2 cytokines by CD4CD25 regulatory T cells in birch allergics during birch pollen season. AB - BACKGROUND: CD4(+)CD25+ regulatory T cells suppress proliferation and cytokine production by human T cells both to self-antigens and exogenous antigens. Absence of these cells in human newborns leads to multiple autoimmune and inflammatory disorders together with elevated IgE levels. However, their role in human allergic disease is still unclear. OBJECTIVE: This study aimed to evaluate the capacity of CD4(+)CD25+ regulatory T cells to suppress proliferation and cytokine production outside and during birch-pollen season in birch-allergic patients relative to non-allergic controls. METHODS: CD4+ cells were obtained from blood of 13 birch-allergic patients and six non-allergic controls outside pollen season and from 10 birch-allergic patients and 10 non-allergic controls during birch pollen season. CD25+ and CD25- fractions were purified with magnetic beads and cell fractions, alone or together in various ratios, were cultured with antigen presenting cells and birch-pollen extract or anti-CD3 antibody. Proliferation and levels of IFN-gamma, IL-13, IL-5 and IL-10 were measured by thymidin incorporation and ELISA, respectively. Numbers of CD25+ cells were analysed by flow cytometry. RESULTS: CD4(+)CD25+ regulatory T cells from both allergics and non-allergics potently suppressed T cell proliferation to birch allergen both outside and during birch-pollen season. However, during season CD4(+)CD25+ regulatory T cells from allergic patients but not from non-allergic controls were defective in down-regulating birch pollen induced IL-13 and IL-5 production, while their capacity to suppress IFN-gamma production was retained. In contrast, outside pollen season the regulatory cells of both allergics and non-allergic controls were able to inhibit T-helper 2 cytokine production. CONCLUSION: This is the first study to show differential suppression of Th1 and Th2 cytokines, with CD4(+)CD25+ regulatory T cells from birch-pollen-allergic patients being unable to down-regulate Th2, but not Th1 responses during birch-pollen season. PMID- 15347369 TI - Aluminium hydroxide down-regulates T helper 2 responses by allergen-stimulated human peripheral blood mononuclear cells. AB - BACKGROUND: Aluminium hydroxide (alum) is a commonly used adjuvant for specific immunotherapy of allergic diseases. While alum is traditionally associated with murine Th2 sensitization, little is known about its effects on secondary allergic responses in humans. METHODS: We investigated the in vitro effects of alum on peripheral blood mononuclear cells (PBMC) from atopic donors. PBMC from 18 grass pollen-sensitive rhinitic subjects were stimulated with Phleum pratense (Phl p) in the presence or absence of alum. After 6 days culture, cytokine production was measured by ELISA and T cell proliferation by radiolabelled thymidine incorporation. The effect of alum on the expression of human leucocyte antigen and CD80/CD86 on cultured antigen-presenting cells was assessed by flow cytometry. RESULTS: PBMC cultured with Phl p and alum showed a significant decrease in both IL-5 and IL-13 production compared with allergen alone (P<0.005 and P<0.001, respectively), but no change in IFN-gamma or IL-12 production or proliferative responses. These alum-induced changes in T helper (Th)2 cytokine production were unaffected by the addition of neutralizing antibodies to IL-4 or IL-12. Culture of PBMC with alum induced increased expression of CD86 (P=0.004) and HLA (P=0.01) on monocytes while the expression of CD80 was decreased (P=0.02). SUMMARY: Alum down-regulates allergen-driven Th2 cytokine responses while Th1 cytokines are unaffected. These data confirm that alum is a useful adjuvant for inclusion in allergen immunotherapy vaccines. PMID- 15347370 TI - CD28 and secretory immunoglobulin A-dependent activation of eosinophils: inhibition of mediator release by the anti-allergic drug, suplatast tosilate. AB - BACKGROUND: Eosinophils are major effector cells in allergic diseases. After their recruitment to sites of inflammation, they contribute to the pathophysiology of the disease by releasing granule proteins and cytokines. Suplatast tosilate (IPD-1151T), a new anti-allergic agent, has shown beneficial effect in the treatment of asthma, associated with reduced bronchoalveolar lavage eosinophil infiltration and eosinophilic cationic protein (ECP) release in serum and sputum. OBJECTIVE: We investigated whether suplatast tosilate could exert direct effects on human eosinophil activation. METHODS: Eosinophils from hypereosinophilic patients or normal donors were purified by Percoll gradient and the magnetic cell separation system. Chemotaxis was studied using the Boyden chamber technique using three chemoattractants, formyl-methionine-leucine phenylalanine (fMLP), IL-5 and eotaxin. Oxidative metabolism was determined by a luminol-dependent chemiluminescence assay after activation with eotaxin or secretory IgA (sIgA). The release of ECP and eosinophil derived neurotoxin (EDN) was measured by radioimmunoassay and cytokine production was determined by ELISA following activation with sIgA or anti-CD28. RESULTS: The chemotactic response to fMLP, IL-5 and eotaxin was significantly inhibited by IPD-1151T. Suplatast tosilate was partially inhibiting the release of reactive oxygen species (ROS) induced by eotaxin and sIgA. Activation by sIgA and CD28 ligation resulted in the release of ECP and EDN, which was inhibited by IPD-1151T. Upon activation by anti CD28, only IL-13 production was inhibited by IPD-1151T, whereas release of IL-2 and IFN-gamma was not affected. IL-10 release induced by sIgA was also inhibited by IPD-1151T. Additionally, the pro-inflammatory cytokine IL-6, which was secreted following anti-CD28 and sIgA stimulation, was strongly inhibited by IPD 1151T. CONCLUSION: Through inhibition of chemotaxis, IPD-1151T might limit the number of eosinophils at the inflammation site. Furthermore, it could reduce the pathological potential of eosinophils by inhibiting the release of ROS and cationic proteins, main inflammatory mediators produced by eosinophils. Moreover, the inhibition of immunoregulatory cytokines released by eosinophils could locally modify the immune response. PMID- 15347371 TI - Effect of montelukast compared with inhaled fluticasone on airway inflammation. AB - BACKGROUND: Inhaled corticosteroids are currently regarded as the gold standard in anti-inflammatory therapy, however, leukotriene receptor antagonists have been ascribed anti-inflammatory properties. OBJECTIVE: We directly compared the anti inflammatory effects of inhaled fluticasone propionate (FP, 100 microg Diskus, twice daily) and oral montelukast (MON 10 mg, nocte) in bronchial biopsies of patients with asthma in a double-blind, double-dummy, parallel-group design. METHODS: Bronchial biopsies, serum and urine samples were collected from 36 atopic asthmatics before and after 8 weeks of treatment. Activated T cells (CD25+), eosinophils (MBP+) and mast cells (tryptase+) were analysed by immunohistochemistry. Serum eosinophil cationic protein (ECP) and IL-5 were analysed by radio and enzyme immunoassay (EIA), respectively. Urinary 9alpha 11beta-PGF2 and leukotriene E4 (LTE4) were measured by EIA. RESULTS: A comparison of changes from baseline [FP/MON ratio (95% confidence interval)] of activated T cells was not different when subjects were treated with FP compared to treatment with MON [1.00 (0.18-4.86); P=0.924]. Following treatment, mast cells in the FP group were significantly lower than in the group treated with MON [0.39 (0.16 0.97); P=0.041]. There was no difference in the number of eosinophils in the lamina propria following either treatment [0.54 (0.05-2.57); P=0.263]. However, treatment with FP resulted in a significantly greater decrease in serum ECP, compared to treatment with MON [0.37 (0.25-0.71); P=0.002]. CONCLUSIONS: FP appears to be superior to MON as an anti-inflammatory therapy in mild asthmatics. PMID- 15347372 TI - Fluticasone propionate aqueous nasal spray does not influence the recurrence rate of chronic rhinosinusitis and nasal polyps 1 year after functional endoscopic sinus surgery. AB - BACKGROUND: Local corticosteroids are widely used in the treatment of nasal polyps and chronic rhinosinusitis both before and after nasal surgery. Their efficacy after functional endoscopic sinus surgery (FESS) has not been fully established by placebo-controlled trials. OBJECTIVE: This double-blind placebo controlled randomized study was performed in order to investigate whether fluticasone propionate aqueous nasal spray (FPANS) reduces the recurrence rate of nasal polyps and chronic rhinosinusitis during the first year after FESS. PATIENTS AND METHODS: The trial looked at 162 patients aged 18 years and older requiring FESS for chronic rhinosinusitis or nasal polyps. After FESS combined with peri-operative systemic corticosteroids, patients were randomized and given FPANS 400 microg b.i.d., FPANS 800 microg b.i.d. or placebo b.i.d. for the duration of 1 year. Patients were withdrawn from the trial (but still included in the study for statistical purposes) if there were recurrent or persistent diseases, defined as progressive regrowth of nasal polyps, recurrent signs and symptoms of chronic sinusitis combined with abnormalities on computed tomography scan and persistent complaints for at least 2 months after FESS. RESULTS: A significant reduction of symptoms was seen after FESS. After 1 year, 46 patients had been withdrawn from the trial because of recurrent diseases and 32 patients because of persistent symptoms. No differences in the number of patients withdrawn because of recurrent or persistent diseases were found between the patients treated with FPANS and patients treated with placebo. We were also unable to find a positive effect of FPANS compared with placebo in several subgroups such as patients with nasal polyps, high score at FESS or no previous sinus surgery. CONCLUSION: This placebo-controlled study does not show that treatment with FPANS up to 1 year after FESS had a positive effect compared with placebo. PMID- 15347373 TI - Desloratadine in combination with montelukast in the treatment of chronic urticaria: a randomized, double-blind, placebo-controlled study. AB - BACKGROUND: Chronic urticaria (CU) is a common skin condition. It is frequently a disabling disease due to the persistency of clinical symptoms, the unpredictable course and negative influence on the quality of life. OBJECTIVE: The aim of this study is to determine whether montelukast, a LTD4 receptor antagonist, plus desloratadine, is more efficacious than desloratadine alone in the treatment of chronic urticaria. MATERIALS: A randomized, double-blind, placebo-controlled study was conducted on 81 patients with a diagnosis of CU. A 1-week single-blind placebo run-in period (baseline) was followed by a 6-weeks double blind active treatment period. The patients were randomized to receive the following treatment once daily: (a) oral desloratadine (5 mg) plus placebo; (b) desloratadine (5 mg) plus montelukast (10 mg); (c) oral placebo alone. The study ended after another 1 week single-blind placebo washout period. RESULTS: The evaluable population thus consisted of 76 patients. Both desloratadine alone and desloratadine plus montelukast administered once daily yielded improvements with respect to the baseline assessment as regards pruritus, number of separate episodes, size and number of weals, visual analogue score and patients' quality of life and with respect to the placebo group both in the active treatment period and in the run out period. However, desloratadine plus montelukast was shown to improve the symptoms and patients' quality of life significantly more than desloratadine alone, although it did not have a significant effect on the number of urticarial episodes. CONCLUSION: The combination of desloratadine plus montelukast is effective in the treatment of CU. It may therefore be a valid alternative in patients with relatively mild CU, in view of its efficacy and the lack of adverse events. PMID- 15347374 TI - Efficacy and safety of subcutaneous immunotherapy with a biologically standardized extract of Ambrosia artemisiifolia pollen: a double-blind, placebo controlled study. AB - BACKGROUND: The allergological relevance of Ambrosia in Europe is growing but the efficacy of the injective immunotherapy for this allergen has been documented only in Northern America. OBJECTIVE: We sought to study the safety and efficacy of injective immunotherapy in European patients sensitized to Ambrosia artemisiifolia. METHODS: Thirty-two patients (18 M/14 F, mean age 36.78, range 23 60 years) suffering from rhinoconjunctivitis and/or asthma and sensitized to Ambrosia were enrolled and randomized in a double-blind, placebo-controlled (DBPC) study lasting 1 year. A maintenance dose corresponding to 7.2 microg of Amb a 1 was administered at 4-week intervals after the build-up. During the second and the third year, all patients were under active therapy in an open fashion. Symptom and medication scores, skin reactivity to Ambrosia (parallel line biological assay), and pollen counts were assessed throughout the trial. RESULTS: Twenty-three patients completed the trial. No severe adverse event was observed. During the DBPC phase, actively treated patients showed an improvement in asthmatic symptoms (P=0.02) and drug (P=0.0068) scores days with asthmatic symptoms (P=0.003), days with rhinitis symptoms (P=0.05), and days with intake of drugs (P=0.0058), as compared to before therapy. No improvement for any of these parameters was detected in the placebo group. Moreover, the number of days with rhinitis and asthma was significantly higher in the placebo as compared to the active group (P=0.048 and P<0.0001, respectively). Patients who switched from placebo to active therapy improved in rhinoconjunctivitis, asthma, and drug intake. The skin reactivity decreased significantly (12.2-fold, P=0.0001) in the active group whereas a slight increase (1.07-fold, P=0.87) was observed in the placebo group after the DBPC phase. After switching to active therapy, patients previously under placebo showed a significant decrease of this parameter (4.78 fold, P=0.002). CONCLUSION: Injective immunotherapy is safe and clinically effective in European patients sensitized to Ambrosia. PMID- 15347375 TI - Prevalence of sensitization to Artemisia allergens Art v 1, Art v 3 and Art v 60 kDa. Cross-reactivity among Art v 3 and other relevant lipid-transfer protein allergens. AB - BACKGROUND: Artemisia vulgaris is a widespread weed in the Mediterranean area and several allergens have been detected in its pollen. One of them, Art v 3, belongs to the lipid-transfer protein (LTP) family and its prevalence in Artemisia sensitized patients or its relationship with other LTP allergens is not clear. OBJECTIVE: To assess the pattern of sensitization to an array of mugwort allergens in a Mediterranean population, and to study the cross-reactivity of Art v 3 with Pru p 3 and Par j 1, relevant LTP allergens in the area. METHODS: Skin prick test was performed with whole extracts (A. vulgaris, Parietaria judaica and peach) and pure natural allergens Art v 1, Art v 3, Art v 60 kDa and Par j 1 in 24 mugwort-allergic patients from a Mediterranean area. In vitro assays included measurement of specific IgE and ELISA inhibition among LTP allergens. RESULTS: The three Artemisia allergens elicited a positive skin response in 70-80% of the patients. Seven patients were clearly sensitized to Par j 1 and 11 to Pru p 3. There was no correlation between Par j 1 and Pru p 3 sensitization, but a highly significant correlation was found between peach extract and Art v 3 as regards the skin response. No IgE cross-reactivity was observed between Art v 3/Par j 1 or Pru p 3/Par j 1. In contrast, Art v 3 significantly inhibited the binding to Pru p 3 of IgE from three patients' sera out of six studied, but Pru p 3 was not able to inhibit the IgE binding to Art v 3. CONCLUSION: Art v 3 is a major mugwort allergen and in some patients with IgE to both Art v 3 and Pru p 3, Art v 3 behaves as the primary sensitizing agent. PMID- 15347376 TI - Mixed antibody and T cell responses to peanut and the peanut allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6 in an oral sensitization model. AB - BACKGROUND: Peanut allergy is known for its severity and persistence through life. Several peanut proteins have been identified as allergenic and are indicated as Ara h 1-7. Very little is known about the mechanisms that underlie sensitization to peanut proteins. OBJECTIVE: The purpose of the present study was to reveal the immune responses that are induced against peanut and the peanut allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6 during sensitization, including the very early responses. METHODS: Humoral and T cell responses against peanut and the peanut allergens were examined in an early and later stage of sensitization in an established murine model of peanut anaphylaxis. Therefore C3H/HeJ mice were orally exposed to two different doses of peanut extract plus cholera toxin. RESULTS: Oral sensitization to peanut was characterized by an antigen-induced mixed cytokine response in the spleen (IL-4, IL-5, IL-10 and IFN gamma), which could already be observed 7 days after the onset of exposure. Additionally, polyisotypic humoral responses (IgE, IgG1 and IgG2a) against peanut were found in the serum. Moreover, we demonstrated that these T helper (Th)1/Th2 cytokine and antibody responses were also directed specifically against the major peanut allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6. CONCLUSIONS: This study implicates that both Th1 and Th2 phenomena are involved in the development of peanut allergy in the C3H/HeJ murine model. Furthermore, we show that the present oral model is suitable to examine immune responses to food allergens during different stages of sensitization upon treatment with a whole food extract. PMID- 15347377 TI - In vitro allergenicity of peanut after hydrolysis in the presence of polysaccharides. AB - BACKGROUND: Peanut is a major allergenic product. Manufacturing processes used in food industries to improve the physicochemical properties of food-based peanut (stabilization, texturization), could cause a modification of the digestibility of peanut proteins and, consequently, their allergenicity. OBJECTIVE: This study aimed at examining the influence of polysaccharides, i.e., gum arabic, low methylated pectin (LMP) and xylan, on the in vitro hydrolysis of peanut protein isolate (PPI) and the in vitro allergenicity of the digestion products. METHODS: PPI was hydrolysed during a two-step in vitro hydrolysis by pepsin, followed by a trypsin/chymotrypsin (T/C) mixture performed in dialysis bags with molecular weight cut-offs (MWCO) of 1000 or 8000 Da. SDS-PAGE electrophoresis and immunoblotting were assessed on the peptic and T/C digestion products in (retentates) and out of the dialysis bags (dialysates). RESULTS: Hydrolysis by all of the digestive enzymes showed retention of some proteins in the dialysis bags in the presence of gum arabic and xylan. The retentates were recognized by IgG and IgE, particularly peptides <20 kDa. The IgE binding with peptides of retentate containing xylan from the dialysis bag with an MWCO of 1000 Da was reduced. The immunoreactivity of hydrolysis products in dialysates was considerably reduced by polysaccharides, regardless of the dialysis bag. CONCLUSION: Reduction of PPI hydrolysis was probably due to non-specific interactions between polysaccharides and peptides. In retentates, IgE-binding epitopes were reduced by digestion and the presence of xylan. In dialysates, they were reduced by all of the polysaccharides. This work highlights the possibility of modulating this food allergy through optimized formulation. PMID- 15347378 TI - A survey of the quality of information leaflets on hayfever available from general practices and community pharmacies. AB - BACKGROUND: Hayfever affects at least one in 10 people. The majority of hayfever is managed in the community setting where the management options are pharmaceutical and behavioural. Hayfever medications are available over the counter and on prescription from the general practitioner. Patient information leaflets are published to augment the advice given and to promote self management, but these leaflets have rarely been subjected to critical review. OBJECTIVE: To assess the quality of patient information leaflets written for people with hayfever and available from general practices and from community pharmacists. METHOD: A structured review of patient information leaflets about hayfever. During the peak grass pollen season copies of all leaflets available were collected from a random sample of community pharmacists and general practices in Wessex. The characteristics of the leaflet were recorded and the contents and presentation of each one was reviewed using the British Medical Association patient information appraisal system. Readability was assessed using the Simple Measure of Gobbledegook (SMOG). Two allergy-accredited specialists assessed each leaflet for accuracy. RESULTS: During the peak pollen season no leaflets were available in 30% of the Community Pharmacists and 23% general practices. In total, 38 different leaflets were identified. All the leaflets reviewed were written for adults. Forty-seven percent of the leaflets had no publication date and one-third of those dated were at least 5 years old. In general the leaflets scored highly on issues of presentation, but less than half contained information on the full range of management and treatment options, many being biased towards a single or limited range of interventions. Seventy-nine percent leaflets were produced or sponsored by pharmaceutical companies and gave prominence to their own products. All the leaflets had readability scores requiring at least secondary education (SMOG score equal or greater than 9). At least one factual inaccuracy was identified in four-fifths of leaflets. CONCLUSION: A large number of leaflets are published for people with hayfever, but they are of variable quality and not accessible to all patients because of limited distribution or high readability scores. If all patients are to benefit from written information the authors and publishers of leaflets need to follow published recommendations, especially with respect to content and readability. In the absence of a national public health information strategy or a body that systematically vets patient information materials, clinicians must pay attention to the quality of information provided for or obtained elsewhere by their patients. PMID- 15347379 TI - Differential effect of mattress covers on the level of Der p 1 and Der f 1 in dust. AB - BACKGROUND: Exposure to house dust mite (HDM) allergens can lead to the development of allergic complaints. Mattress covers seem to be an obvious option for lowering allergen exposure in sensitized individuals. Previous studies have shown that Dermatophagoides pteronissinus was the most prevalent HDM species in the Netherlands. OBJECTIVE: In the present study, we investigated the effect of mattress covers on Der p 1 and Der f 1 concentrations in dust samples in three areas in the Netherlands; Groningen, Utrecht and Rotterdam. METHODS: Dust was obtained from mattresses of 277 patients at the beginning of the study and after 12 months of the placebo-controlled intervention. It was analysed for allergen content by immunoassay. The differential effect of the intervention on Der p 1 vs. Der f 1 was analysed in a subgroup with Der p 1+Der f 1>1 microg/g dust (N=161). It was tested whether the intervention caused a significant change in the Der f 1/Der p 1 ratio. RESULTS: At t=0 we found very similar levels of the group 1 allergens of both species. The relatively high prevalence of D. farinae in our study was geographically restricted: the median Der f 1/Der p 1 ratio was 11.1 in the Rotterdam area compared with 1.32 in the Utrecht area and 0.33 in the Groningen area. Analysis of our data showed that the favourable intervention effect found for the combined allergen data (reduction factor=2.9, P<0.001) is essentially due to a favourable effect of the intervention on the Der f 1 levels only (reduction factor=3.6, P<0.001). The effect on the Der p 1 level was remarkably small (reduction factor: 1.2, P=0.48). In the intervention group, the Der f 1/Der p 1 ratio decreased after 12 months by a factor 2.0, whereas in the placebo group it increased (probability of the intervention effect: P<0.005). CONCLUSION: Mite-impermeable covers are more effective in reducing the level of Der f 1 than that of Der p 1. PMID- 15347380 TI - The flow-cytometric determination of basophil activation induced by aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs) is useful for in vitro diagnosis of the NSAID hypersensitivity syndrome. AB - BACKGROUND: Hypersensitivity reactions to non-steroidal anti-inflammatory drugs (NSAIDs), manifested by cutaneous symptoms and/or airway manifestations represent 20-25% of all hypersensitivity reactions to drugs. Today, it is still claimed that no in vitro diagnostic tests exist for that condition and that the only way to confirm the diagnosis is a provocation challenge. OBJECTIVE: The objective of this study was to assess whether NSAIDs may provoke blood basophil activation in vitro in such patients, as detected by a flowcytometric technique. METHODS: Sixty NSAID hypersensitive patients (38 with cutaneous, 20 with airway and two with cutaneous and airway symptoms) and 30 control patients (15 asthmatics) were selected. Their hypersensitivity was confirmed by documented history indicating at least two clinical episodes to two or more different NSAIDs or by positive oral provocation challenge. Isolated buffy coat leukocytes were stimulated in vitro with aspirin, paracetamol, metamizol, diclofenac, and naproxen. The percentage of activated basophils was evaluated by an anti-CD63. RESULTS: Aspirin showed a sensitivity of 43.3%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value of 99.4%. For the other NSAIDs, the sensitivity and specificity values were: for paracetamol 11.7% and 100%, for metamizol 15% and 100%, for diclofenac 43.3% and 93.3% and for naproxen 54.8% and 74.1%. When considering the first four NSAIDs, the global sensitivity raised to 63.3% and specificity to 93.3%. If the number of tests is to be limited for practical reasons, the combination of acetylsalicylic acid and diclofenac at two concentrations yields a sensitivity of 58.3% and a specificity of 93.3%. CONCLUSIONS: Flowcytometric determinations of basophil activation following stimulation with NSAIDs show a high sensitivity (60-70%) with specificity above 90%. So this test may help avoiding some cumbersome and dangerous provocation challenges. PMID- 15347381 TI - Analysis of effector and regulatory immune reactivity to nickel. AB - BACKGROUND: Diagnostic patch testing in allergic contact dermatitis faces the risk of boosting existing hypersensitivities or active sensitization. Risk-free and reliable in vitro assays using peripheral blood are, therefore, wanted. OBJECTIVES: Here, we studied new approaches for in vitro monitoring of nickel specific effector ad regulatory cell functions in allergic patients and potentially tolerized individuals. METHODS: Lymphocyte proliferation assays were carried out with the allergen and additional IL-12/IL-7 or IL-4/IL-7 cytokine supplements. Release of IFN-gamma and IL-5 were assessed as measures for type-1 and type-2 effector T cell function, respectively, and IL-10 and TGF-beta1 to monitor possible regulatory T cell function reflecting immunological tolerance. After optimization of in vitro cut-off values, potency of these parameters was evaluated as compared with conventional nickel patch testing. RESULTS: One hundred and fifty six outpatients were included in this study, 74 of whom presenting with a positive history of nickel allergy. Nickel-sulphate patch test results showed positive reactions in 43 patients, of whom 40 had a positive history (test sensitivity 54%; specificity 96%; overall accuracy 76%). Proliferation tests without cytokine supplementation showed an accuracy of 68%, which was further improved by supplementing IL-4/IL-7 (82%). IFN-gamma and IL-5 cytokine production, as revealed in IL-12/IL-7 and IL-4/IL-7 supplemented cultures, respectively, showed accuracies of 70% and 83%. As to the production of putatively immunoregulatory cytokines, IL-10 was most informative, with highest production rates in nickel-skin test negative individuals with long-lasting mucosal metal contact preceding skin piercing. CONCLUSIONS: These results indicate that measuring both T cell proliferation and cytokine secretion profiles, in particular IL-5 release using IL-4/IL-7 supplemented medium, offers a promising improvement of the in vitro diagnostic options in monitoring nickel contact sensitization. Since oral nickel contact has been shown earlier to induce active tolerization, nickel-induced in vitro IL-10 production may help identify nickel-tolerized individuals. PMID- 15347382 TI - Increase of CC chemokine receptor 4-positive cells in the peripheral CD4 cells in dogs with atopic dermatitis or experimentally sensitized to Japanese cedar pollen. AB - BACKGROUND: Since dogs frequently develop allergic diseases, similar to those in humans, dogs represent a possible animal model for allergy in humans. In human atopic dermatitis (AD), CC chemokine receptor 4 (CCR4) has been shown to play an important role in the development of allergic inflammation of AD; however, the association between allergic reaction and CCR4 is not well understood in dogs. OBJECTIVE: To examine CCR4 expression in peripheral blood CD4+ cells in dogs that had AD and were experimentally sensitized with Japanese cedar pollen. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from 17 dogs with AD. The proportion of CCR4+ cells in peripheral blood CD4+ cells (CCR4/CD4) was evaluated by flow cytometry and compared with that in 10 healthy dogs. Similarly, in dogs that were experimentally sensitized to Japanese cedar pollen antigen, the proportion of CCR4/CD4 was examined pre- and post sensitization. RESULTS: The proportion of CCR4/CD4 in dogs with AD was 40.3+/ 3.3%, which was significantly higher than that in normal dogs (23.6+/-4.3%) (P<0.01). In the experimentally sensitized dogs, the proportion of CCR4/CD4 was 25.4+/-2.6% at pre-sensitization and it was significantly increased (29.8+/-2.9%) at post-sensitization (P<0.01). CONCLUSION: The proportion of CCR4+ cells in peripheral blood CD4+ cells was measured in dogs with allergic conditions. The present findings indicate that CCR4+ cells may be involved in the pathogenesis of allergy in dogs as in humans. PMID- 15347383 TI - Nerve growth factor-induced substance P in capsaicin-insensitive vagal neurons innervating the lower mouse airway. AB - BACKGROUND: Nerve growth factor (NGF) is elevated in allergic diseases such as bronchial asthma and can lead to an induction of substance P (SP) and related neuropeptides in guinea-pigs large-diameter, neurofilament-positive airway neurons. OBJECTIVE: In the present study, the effect of NGF on tyrosine kinase receptor trkA and the capsaicin receptor TRPV1 expression in airway-specific vagal sensory neurons located in the jugular-nodose ganglia complex (JNC) of mice was investigated. METHODS: Using retrograde neuronal tracing in combination with double-labelling immunohistochemistry, SP, trkA- and TRPV1-receptor expression was examined in airway-specific sensory neurons of BALB/c mice before and after NGF treatment. RESULTS: NGF injected into the lower airway was able to induce SP (13.0+/-2.03% vs. 5.9+/-0.33%) and trkA expression (78+/-2.66% vs. 60+/-2.11%) in larger diameter (>25 microm), capsaicin-insensitive and trkA-positive vagal sensory neurons that were retrograde-labelled with Fast Blue dye from the main stem bronchi. CONCLUSION: Based on the extent of SP and trkA co-expression in airway-specific neurons by NGF treatment, the present study suggests that, following a peripheral activation of trkA receptor on SP afferent by NGF which is elevated in allergic inflammation, there may be trkA-mediated SP induction to mediate neurogenic airway inflammation. PMID- 15347384 TI - Interleukin-1beta attenuates endothelin B receptor-mediated airway contractions in a murine in vitro model of asthma: roles of endothelin converting enzyme and mitogen-activated protein kinase pathways. AB - BACKGROUND: Asthma is a chronic airway disease, known to involve several inflammatory mediators. Little is known about how these mediators interact in order to produce or attenuate even basic features of the disease, like airway hyper-reactivity and remodelling. Endothelin-1 (ET-1) and IL-1beta are two mediators suggested to play important roles in the induction of airway inflammation. OBJECTIVE: To investigate the interactions between ET-1 and IL 1beta, using a novel in vitro model of asthma, focusing on airway smooth muscle contractility. METHODS: Isolated murine tracheal segments were cultured from 1 to 8 days in the absence and presence of IL-1beta. The subsequent contractile responses to sarafotoxin 6c (S6c) (selective agonist for ETB receptor) and sarafotoxin 6b (S6b) (ETA and ETB receptor agonist) were recorded by a myographs system. In all experiments, ETB receptors were desensitized before the contractile response to S6b was recorded. Thus, the response to S6b is only mediated by ETA receptors in the present study. The mRNA expressions for ET-1 and endothelin (ET) receptors were quantified by real-time PCR. RESULTS: Organ culture in the presence of IL-1beta attenuated the maximal contraction induced by S6c, but not S6b. This reduction was concentration-dependent and was significant after 2, 4 and 8 days of culture. To investigate the mechanisms behind this, inhibitors for endothelin converting enzyme (ECE) phosphoramidon, c-JUN N terminal kinase (JNK) SP600125, extracellular-signal-regulated kinase 1/2(ERK 1/2) PD98059 and p38 pathway SB203580 were used. Individually, SP600125 and PD98059, but not SB203580, could partly reverse the reduction induced by IL 1beta. An additional effect was obtained when SP600125 and PD98059 were combined. The mRNA expressions for ET-1 and ETB receptor were up- and down-regulated, respectively, by IL-1beta. CONCLUSION: Presence of IL-1beta in the airways attenuate the contractile response mediated via ETB receptors, an effect dependent on ECE, JNK and ERK 1/2 pathways. PMID- 15347385 TI - Prior Bordetella pertussis infection modulates allergen priming and the severity of airway pathology in a murine model of allergic asthma. AB - BACKGROUND: It has been proposed that T helper (Th)2-driven immune deviation in early life can be countered by Th1 inducing childhood infections and that such counter-regulation can protect against allergic asthma. OBJECTIVE: To test whether Th1-inducing infection with Bordetella pertussis protects against allergic asthma using well-characterized murine models. METHODS: Groups of mice were sensitized to ovalbumin (OVA) in the presence or absence of B. pertussis, a well-characterized Th1 inducing respiratory infection. Immunological, pathological and physiological parameters were measured to assess the impact of infection on immune deviation and airway function. RESULTS: We demonstrate that OVA sensitization does not affect the development of B. pertussis-specific immune responses dominated by IgG2a and IFN-gamma and does not impair Th1-mediated clearance of airway infection. In contrast, B. pertussis infection at the time of sensitization modulated the response to OVA and significantly reduced total serum and OVA-specific IgE. The pattern of cytokine responses, in particular OVA specific IL-5 responses in the spleen was also modulated. However, B. pertussis did not cause global suppression as IL-10 and IL-13 levels were enhanced in OVA stimulated spleen cell cultures and in lavage fluid from infected co-sensitized mice. Histopathological examination revealed that B. pertussis infection prior to OVA sensitization resulted in increased inflammation of bronchiolar walls with accompanying hyperplasia and mucous metaplasia of lining epithelia. These pathological changes were accompanied by increased bronchial hyper-reactivity to methacholine exposure. CONCLUSION: Contrary to the above premise, a Th1 response induced by a common childhood infection does not protect against bronchial hyper reactivity, but rather exacerbates the allergic asthmatic response, despite modulation of immune mediators. PMID- 15347386 TI - Regarding Ebo DG, Hagendorens MM, Bridts CH, Schuerwegh AJ, De Clerck LS & Stevens WJ. In vitro allergy diagnosis: should we follow the flow? Clin Exp Allergy 2004; 34:332-9. PMID- 15347388 TI - Critical review of the vector status of Aedes albopictus. AB - The mosquito Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae), originally indigenous to South-east Asia, islands of the Western Pacific and Indian Ocean, has spread during recent decades to Africa, the mid-east, Europe and the Americas (north and south) after extending its range eastwards across Pacific islands during the early 20th century. The majority of introductions are apparently due to transportation of dormant eggs in tyres. Among public health authorities in the newly infested countries and those threatened with the introduction, there has been much concern that Ae. albopictus would lead to serious outbreaks of arbovirus diseases (Ae. albopictus is a competent vector for at least 22 arboviruses), notably dengue (all four serotypes) more commonly transmitted by Aedes (Stegomyia) aegypti (L.). Results of many laboratory studies have shown that many arboviruses are readily transmitted by Ae. albopictus to laboratory animals and birds, and have frequently been isolated from wild-caught mosquitoes of this species, particularly in the Americas. As Ae. albopictus continues to spread, displacing Ae. aegypti in some areas, and is anthropophilic throughout its range, it is important to review the literature and attempt to predict whether the medical risks are as great as have been expressed in scientific journals and the popular press. Examination of the extensive literature indicates that Ae. albopictus probably serves as a maintenance vector of dengue in rural areas of dengue-endemic countries of South-east Asia and Pacific islands. Also Ae. albopictus transmits dog heartworm Dirofilaria immitis (Leidy) (Spirurida: Onchocercidae) in South-east Asia, south-eastern U.S.A. and both D. immitis and Dirofilaria repens (Raillet & Henry) in Italy. Despite the frequent isolation of dengue viruses from wild-caught mosquitoes, there is no evidence that Ae. albopictus is an important urban vector of dengue, except in a limited number of countries where Ae. aegypti is absent, i.e. parts of China, the Seychelles, historically in Japan and most recently in Hawaii. Further research is needed on the dynamics of the interaction between Ae. albopictus and other Stegomyia species. Surveillance must also be maintained on the vectorial role of Ae. albopictus in countries endemic for dengue and other arboviruses (e.g. Chikungunya, EEE, Ross River, WNV, LaCrosse and other California group viruses), for which it would be competent and ecologically suited to serve as a bridge vector. PMID- 15347389 TI - Borreliae in Ixodes ricinus ticks feeding on humans. AB - A total of 298 Ixodes ricinus (L.) ticks (Acari: Ixodidae) feeding on humans in the Czech Republic were tested for borreliae (Borrelia burgdorferi sensu lato) by darkfield microscopy between 1997 and 2003. A majority (68%) of the supplied I. ricinus ticks were nymphs, 25% were females and 7% were larvae. Overall, 20% of 74 examined females and 9% of 203 examined nymphs (but none of 21 examined larvae) were infected with borreliae. The proportion of ticks with a high infection load (>100 spirochetes) was 4% in females and 2% in nymphal I. ricinus. During the year, the highest numbers and proportions of infected nymphal and female ticks were taken from humans in June. Detection of borreliae in the ticks feeding on humans might be helpful in the prophylaxis of Lyme borreliosis. PMID- 15347390 TI - Sampling fleas: the reliability of host infestation data. AB - The use of measures of host infestation as a reliable indicator of a flea population size to be used in interspecific comparisons was considered. The abundance of fleas collected from host bodies and collected from host burrows was compared among 55 flea species, controlling for the effect of flea phylogeny. The mean number of fleas on host bodies correlated positively with the mean number of fleas in host burrows/nests both when the entire data pool was analysed and for separate subsets of data on 'fur' fleas and 'nest' fleas. This was also true for a within-host (Microtus californicus) between-flea comparison. The results of this study demonstrate that, in general, the index of host body infestation by fleas can be used reliably as an indicator of the entire population size. PMID- 15347391 TI - Detection of Campylobacter and Escherichia coli O157:H7 from filth flies by polymerase chain reaction. AB - Flies (Diptera: Muscidae) that breed in faeces and other organic refuse (filth flies) have been implicated as vectors of pathogenic bacteria including Escherichia coli O157:H7, which cause haemorrhagic colitis in humans, and Campylobacter, which is the principal causative agent of human enteritis. The potential role of filth flies in the epidemiology of these pathogens in the United States was investigated by examining the prevalence of Campylobacter spp. and E. coli O157:H7 from two Arkansas turkey facilities. Polymerase chain reaction was conducted on DNA extractions of individual Musca domestica Linnaeus, Stomoxys calcitrans (Linnaeus), Hydrotaea aenescens (Wiedemann), Adia cinerella Fallen and turkey faecal samples using primers specific for E. coli H7, O157 and Campylobacter spp. Culturing verified that the flies were carrying viable Campylobacter spp. and E. coli O157:H7. Results from this study indicated that M. domestica, S. calcitrans, H. aenescens and Anthomyids are capable of carrying Campylobacter in North American poultry facilities and that the E. coli O157:H7 is carried by house flies and black dump flies associated with poultry. This PCR method provided a rapid and effective method to identify Campylobacter spp. and E. coli O157:H7 directly from individual filth flies. PMID- 15347392 TI - Exposure of sheep to mosquito bites: possible consequences for the transmission risk of Rift Valley Fever in Senegal. AB - Rift Valley Fever (RVF) is a growing health problem in West Africa. In northern Senegal, the candidate vectors of this arbovirosis are Aedes (Aedimorphus) vexans Meigen and Culex (Culex) poicilipes Theobald (Diptera: Culicidae). Domestic ruminants are the reservoirs of the virus. A study was undertaken during the 2002 rainy season to assess spatial and temporal variations in exposure to mosquito bites in sheep herds, and to evaluate the possible consequences on the risk of RVF transmission to sheep. Mosquitoes were collected with sheep-baited traps. The number of Ae. vexans females (the predominant species during the 2002 rainy season) trapped per trap-night was the dependent variable in statistical analyses. The trapping periods were divided into six series of two to five consecutive days, from July to November 2002. Three temporary ponds were selected according to their ecological features: depth, bank slope, size and vegetation cover. Traps were laid on the pond bank and in the nearest available compound, close to the sheep night pen. Data were analysed using mixed-effects Poisson models. The explanatory variables were the trapping period, the pond, and the capture site. The exposure to mosquito bites varied according to the pond type, suggesting that the risk of transmission was spatially heterogeneous. However, there was no obvious trend in transmission risk due to the effect of the distance from the compound to the pond. The period with the highest exposure was in October, i.e. when transhumant herds left the Ferlo to relocate to their dry season settlement. It is thus hypothesized that transhumance, the seasonal movements of herds, plays a significant role in the dissemination of RVF virus in the region. PMID- 15347393 TI - Biological diversity versus risk for mosquito nuisance and disease transmission in constructed wetlands in southern Sweden. AB - In southern Sweden, many wetlands have been constructed, and maintaining or increasing biological diversity is often included in the aims. Some wetlands are constructed near human settlements, thus raising the problem of wetlands being associated with mosquitoes (Diptera: Culicidae). Increased biodiversity (including mosquito diversity) is considered desirable, whereas mosquito nuisance from a human point of view is not. Adult mosquito abundance, diversity and species assemblages of constructed wetlands were compared to natural wetlands. The potential of constructed wetlands for mosquito nuisance and transmission of mosquito-borne viruses was evaluated. The study areas included five constructed and four natural wetlands. Mosquito abundance and species richness were higher in the natural than in the constructed wetlands, and showed a positive correlation with wetland size. Mosquito species assemblages formed three clusters, which were not explained by origin, size and water permanence of wetlands. In a redundancy analysis, however, mosquito faunas showed significant relationships with these variables, and size and origin of wetlands were most important. Major nuisance species (multivoltine species feeding on mammals and laying eggs on soil) were found in all wetlands, although in relatively low numbers. Risk assessment for Sindbis virus transmission showed moderate risk for two constructed wetlands near human settlements. It is concluded that small size of constructed wetlands has the advantage of low mosquito numbers from a human point of view. The use of functional groups is recommended as a tool for presenting mosquito data to the public, and for helping communication between scientists and administrative decision makers. PMID- 15347394 TI - Susceptibility of mosquito and tick cell lines to infection with various flaviviruses. AB - The genus Flavivirus consists of more than 70 virus species and subtypes, the majority of which are transmitted by mosquitoes or ticks, although some have no known vector (NKV). The ability of these viruses to infect cultured cells derived from mosquito or tick species offers a useful insight into the suitability of such vectors to harbour and replicate particular viruses. We undertook a comparative study of the susceptibility of mammalian Vero cells, a clonal mosquito cell line (C6/36) and recently developed cell lines derived from the ticks (Acari: Ixodidae) Ixodes ricinus (L.) (IRE/CTVM18), I. scapularis (Say) (ISE6), Rhipicephalus appendiculatus (Neumann) (RAE/CTVM1) and Amblyomma variegatum (Fabricius) (AVL/CTVM17) to infection with 13 flaviviruses (and one alphavirus) using immunofluorescence microscopy and plaque assay techniques. The C6/36 mosquito cell line was infected by all the mosquito-borne flaviviruses tested but not by NKV viruses or tick-borne viruses, with the exception of Langat virus (LGTV). The tick cell lines were susceptible to infection by all of the tick-borne viruses tested, as well as two mosquito-borne viruses, West Nile virus (WNV) and the alphavirus, Venezuelan equine encephalitis virus (VEEV), but not other mosquito-borne viruses or NKV viruses. PMID- 15347395 TI - Variation in the load of the horn fly, Haematobia irritans, in cattle herds is determined by the presence or absence of individual heifers. AB - The distribution of horn flies, Haematobia irritans (L.) (Diptera: Muscidae), in herds of Danish Holstein-Friesian cattle was investigated in two studies conducted during two field seasons. In the first study, highly significant differences in fly distribution between the most and the least fly-susceptible heifers were observed. In one herd, the mean difference between the most fly susceptible and the most fly-resistant heifers was 268 Ha. irritans specimens. The highest ratio between upper and lower mean fly number was 64.1:1, whereas the lowest was 3.1:1. In the second year, it was demonstrated that the heifers kept their rank in fly attraction over time. The trial clearly demonstrated that some heifers were attracting flies, whereas others, even in the same herd, only carried a few. In the second study, heifers were moved in and out of herds in an attempt to manipulate fly loads in the herds. In year 1, one herd (herd A) received four fly-resistant heifers from another herd (herd B), resulting in a drop in the mean number of flies, whereas herd B received four fly-susceptible heifers from herd A, resulting in an elevation of the mean number of flies. In year 2, a similar pattern emerged using herds C and D, and when the cattle were later returned to their original herds, the fly loads returned to their original distribution. The data presented here show unequivocally that, for horn flies, there can be considerable differences in fly loads for individual heifers within the Holstein-Friesian breed. Furthermore, the overall fly load within herds can be manipulated, and can be reversed. Thus, the distribution in the number of flies within a herd appears to depend on the number of fly-resistant or fly susceptible heifers. The possible role of chemical factors emitted by heifers, i.e. volatile semiochemicals, in determining differences in fly loads is discussed, whereby attractants are emitted by fly-susceptible heifers and enable flies to locate their host, and repellents are emitted by fly-resistant heifers such that the flies are actively repelled from the herd. PMID- 15347396 TI - Flavivirus isolations from mosquitoes collected from Saibai Island in the Torres Strait, Australia, during an incursion of Japanese encephalitis virus. AB - Adult mosquitoes (Diptera: Culicidae) were collected in January and February 2000 from Saibai Island in the Torres Strait of northern Australia, and processed for arbovirus isolation during a period of Japanese encephalitis (JE) virus activity on nearby Badu Island. A total of 84 210 mosquitoes were processed for virus isolation, yielding six flavivirus isolates. Viruses obtained were single isolates of JE and Kokobera (KOK) and four of Kunjin (KUN). All virus isolates were from members of the Culex sitiens Weidemann subgroup, which comprised 53.1% of mosquitoes processed. Nucleotide sequencing and phylogenetic analysis of the pre-membrane region of the genome of JE isolate TS5313 indicated that it was closely related to other isolates from a sentinel pig and a pool of Cx. gelidus Theobald from Badu Island during the same period. Also molecular analyses of part of the envelope gene of KUN virus isolates showed that they were closely related to other KUN virus strains from Cape York Peninsula. The results indicate that flaviviruses are dynamic in the area, and suggest patterns of movement south from New Guinea and north from the Australian mainland. PMID- 15347397 TI - Mitochondrial diversity analysis of Glossina palpalis gambiensis from Mali and Senegal. AB - West African riverine tsetse populations of Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) were investigated for gene flow, inferred from mitochondrial diversity in samples of 69 flies from Senegal and 303 flies from three river drainages in Mali. Four polymorphic mitochondrial loci were scored. Mean haplotype diversities were 0.30 in Mali, 0 in Senegal and 0.18 over both Mali and Senegal. These diversities estimate the probabilities that two randomly chosen tsetse have different haplotypes. Substantial rates of gene flow were detected among flies sampled along tributaries belonging to the river basins of the Senegal, Niger, and Bani in Mali. There was virtually no gene flow between tsetse in Senegal and Mali. No seasonal effects on gene flow were detected. The implications of these preliminary findings for the implementation of area-wide integrated pest management (AW-IPM) programmes against riverine tsetse in West Africa are discussed. PMID- 15347398 TI - A new cytotype of Simulium squamosum from south-west Cameroon. AB - A new cytotype of Simulium squamosum Enderlein (Diptera: Simuliidae) (here named S. squamosum'D') is described from around Mount Cameroon in south-west Cameroon on the basis of sex-chromosome differences on the short arm of chromosome I. Two polymorphic inversions, IS-To (a new inversion) and IS-2, are partially Y linked. These inversions usually occur together, although IS-To has a higher frequency and is more strongly Y linked than IS-2. This sex-chromosome system has not been previously described and the occurrence and evolution of sex-chromosome systems in S. squamosum is discussed. PMID- 15347399 TI - Habitat characteristics of Anopheles gambiae s.s. larvae in a Kenyan highland. AB - Anopheline larval habitats associated with a swamp, were examined in a highland area (1910 m elevation) of western Kenya. A significant association was found between occurrence of Anopheles gambiae Giles s.s. (Diptera: Culicidae) larvae and two factors, habitat size and vegetation type. Over 80% of An. gambiae s.s. larvae were found in small isolated pools, characterized by short plants, occurring in both swamp margins and roadside ditches. However, Anopheles gambiae s.s. was not found in habitats marked by papyrus and floating plants. The larval habitat of An. gambiae s.s. was characterized by warmer daytime temperatures of water, which were significantly affected by habitat size and plant size. The density of indoor resting An. gambiae s.s. was 0.22 per house and negatively associated with distance from the swamp. These results indicate that the practice of swamp cultivation, in populated areas of the African highlands, increases availability and enhances habitat conditions for the malaria vector. PMID- 15347400 TI - Infestation of the human intestine by the millipede, Nopoiulus kochii. AB - Specimens of the millipede, Nopoiulus kochii (Gervais) were seen in faeces and vomit of a 14-year-old boy residing in Oltu, Erzurum, Turkey. The patient complained of a burning sensation in his throat and stomach-ache. Physical examination revealed no pathological findings, and how the patient became infected was unknown. Anti-parasitic drugs (niclosamide, albendazole), which he had been taking intermittently for 2 years by the prescription of a physician, had not resolved the problem. PMID- 15347401 TI - Records of Chrysomya albiceps in the Ukraine. AB - First records of the presence of the blowfly, Chrysomya albiceps in the western and northern Ukraine are presented. PMID- 15347403 TI - Beyond publication. PMID- 15347405 TI - Responses to the spiritual needs of older people. AB - BACKGROUND: The literature suggests that the notion of holistic health has gained popularity in the nursing of older persons. Holistic care, based on the premises that there is a balance between body, mind and spirit, is important for well being, that each of these is interconnected, and that each affects the others. Human spirit is considered to be the essence of being and is what motivates and guides us to live a meaningful existence. However, there is little evidence in the nursing literature about how nurses caring for older people respond to their spiritual needs. AIM: The aim of this paper is to report a critical incident study to: (1) explore nurses' perceptions of their role in addressing the spiritual needs of older people; (2) describe what constitutes spiritual care of old people in the light of the findings. METHODS: Descriptions of critical incidents were obtained from a convenience sample of 52 nurses working in the East Midlands Region of the United Kingdom (UK) and subjected to content analysis and construction of a data classification system. FINDINGS: Respondents were prompted to identify patients' spiritual needs by factors such as religious beliefs and practice (prayer); absolution; seeking connectedness, comfort and reassurance, healing or searching for meaning and purpose. The interventions initiated to meet patients' spiritual needs included respect for privacy; helping patients to connect; helping patients to complete unfinished business; listening to patients' concerns; comforting and reassuring; using personal religious beliefs to assist patients and observation of religious beliefs and practices. CONCLUSION: The findings provide empirical evidence of some practices related to spiritual care of older people. Further empirical research is needed to guide practice and education with regard to conceptual clarity and the delivery of spiritual care of older people. PMID- 15347406 TI - Community nursing: health professional and public perceptions. AB - BACKGROUND: Almost 10 years ago senior United Kingdom nurses met at Heathrow Airport to discuss the challenges and opportunities facing nursing. One aspect of this 'Heathrow Debate' was to encourage nurses to work closer with members of the public. The resulting report stated that nurses have not been effective in working outside their professional area and, in particular, they have not been adept at working in partnership with the public. AIM: The aim of this paper is to report a study to obtain the views of general practitioners (GPs), community nurses and members of the public on how the public perceive community nursing. A further aim was to explore the perceptions of senior health care managers and policymakers about public involvement in primary care. METHODS: A two-stage Delphi investigation was undertaken with community nurses (n = 38), GPs (n = 14) and members of the public (n = 8). In addition, one-to-one interviews were carried out with 34 senior health service managers and policymakers. FINDINGS: There was agreement on the following issues: community nurses must work more closely with members of the public; there is a lack of public understanding on the roles of community nurses and specialist nurses; there is public support for nurse prescribing; there is a preference for seeing the community nurse as opposed to the GP; and people prefer one nurse to care for them rather than a variety of nurses. Comments from managers and senior policymakers showed agreement on the public being involved in primary care, but an acknowledgement that this was problematic. A number of recommendations are outlined. PMID- 15347407 TI - Protocolized weaning from mechanical ventilation: ICU physicians' views. AB - BACKGROUND: The use of protocols during weaning from mechanical ventilation is uncommon in the UK, despite research pointing to their potential benefits. This may be because the research evidence is considered not to apply in different settings. Intensive care unit consultant physicians are the major decision-makers in weaning in the UK and any attempt to introduce protocolized weaning will require consideration of their views. AIM: The aim of this paper is to report a study exploring intensive care physicians' views on (i) weaning from mechanical ventilation, (ii) the utility of weaning protocols and (iii) nurses' roles in the weaning process. A specific goal was to identify potential aids and barriers to developing weaning protocols and their introduction into clinical practice. METHODS: Qualitative interviews were conducted with a purposive sample of 10 consultant physicians in two intensive care units in Northern Ireland and subjected to content analysis. FINDINGS: The primary themes identified were (i) information required for weaning decisions and clinical judgement, (ii) professional boundaries, (iii) protocol issues and (iv) timing of weaning. Three types of information were deemed to be required for weaning decisions - empirical objective, empirical subjective and abstract - and interviewees considered that it would be challenging to incorporate all into a protocol. They were divided on whether protocols were useful when nursing experience was limited. Some groups of patients were thought more suitable than others for protocolized weaning. CONCLUSIONS: Although local physicians were supportive in theory, introduction of protocolized weaning is likely to be difficult because of the breadth of information required for successful decision-making. Consultant views in this study were not consistent with American findings that physicians' caution may unnecessarily prolong weaning. PMID- 15347408 TI - Coping strategies used by the relatives of people with obsessive-compulsive disorder. AB - BACKGROUND: Obsessive-compulsive disorder (OCD) is a destructive mental illness that alters the lives of both patients and their relatives. Many investigations have described the coping strategies of relatives of patients with schizophrenia, depression and other psychiatric disorders, but there have been no studies reported about coping strategies and OCD. AIM: The aim of this paper is to report an investigation into experiences of burden in relatives of patients with OCD, and the coping strategies they had developed. METHOD: Narrative interviews with 22 family members of patients with OCD were analysed using a grounded theory approach. FINDINGS: Relatives described different burdens and developed different strategies to cope with these. While parents tried to educate their ill children, spouses focused on the patient's resources. The attempts of family members to cope with patients with OCD included assisting in rituals, opposing the symptoms and supporting patients in dealing with the illness. CONCLUSION: Implications for clinical practice to support these relatives are discussed. PMID- 15347409 TI - Changing nurses' dis-empowering relationship patterns. AB - BACKGROUND: Nurses' inter- and intra-disciplinary relationships are frequently interpreted as oppressed group behaviours, contributing to their relatively dis empowered status. In the context of restructuring in health care, this study examined the views of hospital staff nurses about their relationships with nursing colleagues and other health care professionals and their ideas for change. AIM: The aim of this paper is to report a study to add the views of staff nurses to the discourse on restructuring and to make visible the processes that contribute to their marginalization. DESIGN: The study was descriptive and exploratory. Staff nurses from various hospitals in a large Canadian city were selected by theoretical sampling. METHOD: Twenty volunteer staff nurses were interviewed between 1998 and 1999, using broad, open-ended questions and prompts to explore nurses' various relationships in the health care system. This approach allowed for multiple responses and expansions of ideas, without losing focus. The interviews were audio-taped and later transcribed. Thematic analysis was carried out. FINDINGS: Many participants were aware of inter-disciplinary hierarchies, particularly between nurses and physicians. Many also showed insights into their own intra-disciplinary hierarchies and mutual non-supportiveness. Both types of relationships were found to be inextricably linked, sustaining nurses' oppression through dis-empowering discourses. Nurses expressed many ideas about how to promote mutually supportive relationships. CONCLUSION: Change for the better needs to come from within the nursing profession. To develop effective strategies, bedside nurses have to be included in decision-making processes affecting them and their practice, about which they are the experts. Mutual respect, awareness-raising through education, development of caring nursing communities, mentorship and non-hierarchical leadership are key to stopping dis empowering discourses and practices amongst nurses. PMID- 15347410 TI - Meeting the needs of minority ethnic patients. AB - BACKGROUND: Certain aspects of nursing care related to minority ethnic patients are being highlighted in the literature, but there is little exploration of nurses' experiences of caring for people from specific minority ethnic groups. AIM: This paper reports an investigation into the experiences of Registered Nurses caring for hospitalized Pakistani patients in the United Kingdom. METHOD: A qualitative study, with a sample of 30 Registered Nurses using semi-structured interviews. The sample was self-selecting from a large health care organization in the north of England, covering adult acute, critical and rehabilitation care settings. RESULTS: Interviewees had difficulty in explaining the meaning of culture and spirituality and their relationship to nursing practice. They also had limited understanding of the Pakistani community, and deficits were identified in meeting the challenges offered by this community. Inadequate implementation of 'holism', poor preparation to meet the needs of an ethnically diverse society and the presence of racism in practice settings emerged as explanations for the deficits participants identified between their expectations and the reality in care settings. CONCLUSION: Although 'holism' is a relevant concept for enhancing nursing practice, its meaning needs to be further debated in order to avoid a tokenistic approach to its implementation in the care of patients from minority ethnic communities. PMID- 15347411 TI - Emergency care and narrative knowledge. AB - BACKGROUND: Nursing knowledge is a composite of many influences. In an attempt to strengthen nursing knowledge and practice, there have been significant developments in the codification of nursing knowledge as part of the evidence based practice movement. Using established research techniques; this has the capacity to enhance clinical practice. However, not all knowledge can be incorporated into this type of academic discourse, and there is growing interest in narrative knowledge. This paper contributes to this sometimes implicit and sometimes explicit critique of dominant paradigms. AIM: The aim of this paper is to examine the usefulness of narrative as a means of exploring the world of emergency nursing practice and its contribution to the emotional lifeworld of clinicians. METHOD: A discourse analysis of nursing narratives in their natural settings was carried out. This involved exploration of stories that were not produced for the purposes of research. FINDINGS: Narratives can open up social worlds, which escape or be deliberately erased by more formal methods. Contingent and subversive knowledge can contribute to understanding the emotional impact of emergency care. CONCLUSION: These narratives have their own 'truth', which should be incorporated into an understanding of what constitutes the lifeworld of emergency nurses. PMID- 15347412 TI - Diabetes care: practice nurse roles, attitudes and concerns. AB - BACKGROUND: Practice nurses (PNs) are the largest group of nurses providing primary care for patients with diabetes in New Zealand, and changes in the health system are likely to have a substantial effect on their roles. To inform the development of a new primary health care nursing structure and evaluate the new role associated with this, it will be important to have data on current practice nurse roles. AIMS: The aim of this paper is to report a study to compare the diabetes-related work roles, training and attitudes of practice nurses in New Zealand surveyed in 1990 and 1999, to consider whether barriers to practice nurse diabetes care changed through that decade, and whether ongoing barriers will be addressed by current changes in primary care. METHODS: Questionnaires were mailed to all 146 PNs in South Auckland in 1990 and to all 180 in 1999, asking about personal and practice descriptions, practice organization, time spent with patients with diabetes, screening practices, components of care undertaken by practice nurses, difficulties and barriers to good practice, training in diabetes and need for further education. The 1999 questionnaire also asked about nurse prescribing and influence on patient quality of life. RESULTS: More nurses surveyed in 1999 had postregistration diabetes training than those in 1990, although most of those surveyed in both years wanted further training. In 1999, nurses looked after more patients with diabetes, without spending more time on diabetes care than nurses in 1990. Nevertheless, they reported increased involvement in the more complex areas of diabetes care. Respondents in 1999 were no more likely than those in 1990 to adjust treatment, and gave a full range of opinion for and against proposals to allow nurse prescribing. The relatively low response rate to the 1990 survey may lead to an underestimate of changes between 1990 and 1999. CONCLUSIONS: Developments in New Zealand primary care are likely to increase the role of primary health care nurses in diabetes. Research and evaluation is required to ascertain whether this increasing role translates into improved outcomes for patients. PMID- 15347413 TI - A concept analysis of dignity for older adults. AB - BACKGROUND: Human dignity is an essential value of professional nursing education as well as a component of the American Nurses Association Code of Ethics. Nurses are exhorted to treat patients with dignity, and older adults want to be treated with dignity and die with dignity. Although dignity, particularly the dignity of older adults, is often discussed in the health care literature, its meaning is not always clear. AIM: The aim of this paper is to describe a concept analysis to develop a definition of dignity in older adults. METHODS: Data were collected using a literature review and five focus groups composed of older adults. The literature provided data about professionals' ideas of dignity and the focus groups provided qualitative data about the nature of dignity in older people. The literature review and focus groups were carried out concurrently, followed by synthesis of the findings. FINDINGS: Dignity is an inherent characteristic of being human, it can be subjectively felt as an attribute of the self, and is made manifest through behaviour that demonstrates respect for self and others. Dignity must be learned, and an individual's dignity is affected by the treatment received from others. CONCLUSIONS: A behavioural definition of dignity was constructed and this could provide the theoretical basis for nurses to develop interventions that foster dignity for older people. PMID- 15347414 TI - The development of evidence-based nursing interventions: methodological considerations. AB - BACKGROUND: The call for evidence-based practice presents numerous challenges to nurses who are responsible for developing interventions and expanding the associated knowledge base. The challenge is compounded because there is limited literature concerning development of interventions and their evidence base. AIM: The purpose of this article is to present a model that has been successfully used to guide the process of developing and testing complex nursing interventions, especially those in which the experience of the client plays an important role. DISCUSSION: The model consists of four stages: problem definition, accumulation of building blocks for intervention design, intervention design and intervention validation. Each stage is described and examples from research studies are presented. Specific attention is given to the manner in which the model allows for the accumulation of empirical evidence and theory development during the development process. CONCLUSIONS: Use of the model could facilitate effective communication among nurses, researchers and educators when discussing the development and testing of nursing interventions. PMID- 15347415 TI - Aromatherapy practice in nursing: literature review. AB - BACKGROUND: The use of aromatherapy in nursing care continues to be popular in many settings. Most of the nursing literature relates to the use of essential oils in low doses for massage or use of the oils as environmental fragrances. Information from the wider literature may add to the evidence base for use of this therapy in nursing. AIM: This paper reports a literature relating to the use of aromatherapy by nurses and critically evaluates the evidence to support this practice. METHOD: Medline, CINAHL, MANTIS and EBSCO Host databases were searched for papers related to use of essential oils and/or aromatherapy. Papers were also obtained through cross-checking of reference lists. A total of 165 articles have been included in this review. Nursing papers were published since 1990 were included, but some references from 1971 onwards relating to scientific research conducted on essential oils were also included. These remain valuable as they are probably the only reference available for a specific oil or property, or show the development of knowledge in this area. Papers were excluded if they consisted only of brief case studies presented in abstract form. The review covers key professional issues and the principal areas of clinical practice where aromatherapy is used. FINDINGS: Despite calls for more research in the 1980s and 1990s, there is still little empirical evidence to support the use of aromatherapy in nursing practice beyond enhancing relaxation. Its popularity needs to be balanced against the potential risks related to allergies, safety and inappropriate use by inexperienced users. CONCLUSIONS: There is great potential for more collaborative research by nurses to explore the clinical applications in greater detail and to move beyond the low dose paradigm of application of essential oils. PMID- 15347416 TI - Tuberculosis levels ten times higher in Eastern Europe and Central Asia. PMID- 15347421 TI - Mohmmar Qadaffi, open access, and retrovirology. AB - Retrovirology has been publishing as an Open Access online journal for approximately six months. In this editorial, I review the reasons for and the advantages of Open Access publishing, update our progress to date, and summarize where we intend to go with this journal. PMID- 15347422 TI - Daily rhythms in plasma levels of homocysteine. AB - BACKGROUND: There is accumulated evidence that plasma concentration of the sulfur containing amino-acid homocysteine (Hcy) is a prognostic marker for cardiovascular morbidity and mortality. Both fasting levels of Hcy and post methionine loading levels are used as prognostic markers. The aim of the present study was to investigate the existence of a daily rhythm in plasma Hcy under strictly controlled nutritional and sleep-wake conditions. We also investigated if the time during which methionine loading is performed, i.e., morning or evening, had a different effect on the resultant plasma Hcy concentration. METHODS: Six healthy men aged 23-26 years participated in 4 experiments. In the first and second experiments, the daily rhythm in Hcy as well as in other amino acids was investigated under a normal or an inverse sleep-wake cycle. In the third and fourth, Hcy concentrations were investigated after a morning and evening methionine loading. To standardize food consumption in the first two experiments, subjects received every 3 hours 150 ml of specially designed low protein liquid food (Ensure(R) formula). RESULTS: In both the first and second experiments there was a significant daily rhythm in Hcy concentrations with a mid day nadir and a nocturnal peak. Strikingly different 24-h patterns were observed in methionine, leucine, isoleucine and tyrosine. In all, the 24-h curves revealed a strong influence of both the sleep-wake cycle and the feeding schedule. Methionine loading resulted in increased plasma Hcy levels during both morning and evening experiments, which were not significantly different from each other. CONCLUSIONS: There is a daily rhythm in plasma concentration of the amino acid Hcy, and this rhythm is independent of sleep-wake and food consumption. In view of the fact that increased Hcy concentrations may be associated with increased cardiovascular risks, these findings may have clinical implications for the health of rotating shift workers. PMID- 15347423 TI - Dichotomous factor analysis of symptoms reported by UK and US veterans of the 1991 Gulf War. AB - BACKGROUND: Factor analysis is one of the most used statistical techniques to analyze the inter-relationships among symptoms reported by Gulf War veterans. The objective of this study was to apply factor analyses to binary symptom data from the UK study of Gulf War illness and the US Air Force study of Gulf War veterans, and to compare the symptom domains derived from the distinct samples. METHODS: UK veterans of the 1991 Gulf War (n = 3,454), individuals deployed to Bosnia on U.N. peacekeeping operations (n = 1,979) and Gulf War-era servicemen (n = 2,577) who were not deployed to the Gulf were surveyed in 1997-1998, and US 1991 Gulf War veterans from four Air Force units (n = 1,163) were surveyed in 1995 to collect health characteristics including symptoms. Each sample was randomly split in half for exploratory and confirmatory dichotomous factor analyses with promax oblique rotation. RESULTS: Four correlated factors were identified in each of the samples. Three factors (Respiratory, Mood-Cognition, Peripheral Nervous) overlapped considerably across the UK cohorts. The Gastrointestinal/Urogenital factor in the UK Gulf cohort was noticeably different from the Gastrointestinal factor identified from the Bosnia and Era cohorts. Symptoms from Gulf War UK and U.S cohorts yielded similar Gastrointestinal, Respiratory and Mood-Cognition factors, despite differences in symptom inventories between the two surveys. A Musculoskeletal factor was only elicited from the US Gulf sample. CONCLUSION: Findings of this report are consistent with those from other factor analysis studies that identified similar symptom dimensions between Gulf and non-Gulf War veterans, except that the Gastrointestinal factor in Gulf veterans included other symptom types. Correlations among factors raise the question as to whether there is a general illness, even if not unique to Gulf veterans, representing the common pathway underlying the identified factors. Hierarchical factor analysis models may be useful to address this issue. PMID- 15347424 TI - Polyphosphoinositides suppress the adhesion of Haemophilus influenzae to pharyngeal cells. AB - BACKGROUND: One of the primary causes of otitis media (OM), an inflammation of the middle ear, is the bacterium Haemophilus influenzae (HI). OM often occurs to young children, and is mostly treated with antibiotics. Due to concerns over bacterial resistance toward antibiotics, reliable prophylactic treatments such as administrating anti-adhesion agents are now viewed as viable alternatives. RESULTS: The present study tested the feasibilty of using phosphoinositides as anti-adhesion agents against HI cells. Cells of non-typeable HI were radiolabeled with 111- indium-oxine, pre-incubated with various individual phosphoinositides for 15 minutes at 37 degrees C, and incubated with a monolayer of human pharynx carcinoma (DT 562) cells for 20 minutes at 37 degrees C. The result showed that at 0.1 mg/mL dipalmitoylphosphatidylinositol-3,4-diphosphate (PI-3,4-PP) had the highest anti-adhesion activity, followed by phosphatidylinositol-3-phosphate (PI 3-P) and phosphatidylinositol-4-phosphate (PI-4-P). The anti-adhesion activity of PI-3,4-PP was dose-dependent ranging from 0.006 to 0.1 mg/mL. In addition, results from an in vivo study demonstrated that pre-incubation of HI cells with PI-3,4-PP at 1 mg/mL suppressed the growth of HI in nasopharynx of neonatal rats. CONCLUSIONS: These findings suggest that PI-3-P and PI-4-P and more so PI-3,4-PP may serve as prophylactic agents against HI adhesion and colonization. PMID- 15347425 TI - Recombinant antigen-based antibody assays for the diagnosis and surveillance of lymphatic filariasis - a multicenter trial. AB - The development of antifilarial antibody responses is a characteristic feature of infection with filarial parasites. It should be possible to exploit this fact to develop tools to monitor the progress of the global program to eliminate lymphatic filariasis (LF); however, assays based on parasite extracts suffer from a number of limitations, including the paucity of parasite material, the difficulty of assay standardization and problems with assay specificity. In principle, assays based on recombinant filarial antigens should address these limitations and provide useful tools for diagnosis and surveillance of LF. The present multicenter study was designed to compare the performance of antibody assays for filariasis based on recombinant antigens Bm14, WbSXP, and BmR1. Coded serum specimens were distributed to five participating laboratories where assays for each antigen were conducted in parallel. Assays based on Bm14, WbSXP, or BmR1 demonstrated good sensitivity (>90%) for field use and none of the assays demonstrated reactivity with specimens from persons with non-filarial helminth infections. Limitations of the assays are discussed. Well-designed field studies are now needed to assess sampling methodology and the application of antibody testing to the monitoring and surveillance of LF elimination programs. PMID- 15347426 TI - Improving prescribing of antihypertensive and cholesterol-lowering drugs: a method for identifying and addressing barriers to change. AB - BACKGROUND: We describe a simple approach we used to identify barriers and tailor an intervention to improve pharmacological management of hypertension and hypercholesterolaemia. We also report the results of a post hoc exercise and survey we carried out to evaluate our approach for identifying barriers and tailoring interventions. METHODS: We used structured reflection, searched for other relevant trials, surveyed general practitioners and talked with physicians during pilot testing of the intervention. The post hoc exercise was carried out as focus groups of international researchers in the field of quality improvement in health care. The post hoc survey was done by telephone interviews with physicians allocated to the experimental group of a randomised trial of our multifaceted intervention. RESULTS: A wide range of barriers was identified and several interventions were suggested through structured reflection. The survey led to some adjustments. Studying other trials and pilot testing did not lead to changes in the design of the intervention. Neither the post hoc focus groups nor the post hoc survey revealed important barriers or interventions that we had not considered or included in our tailored intervention. CONCLUSIONS: A simple approach to identifying barriers to change appears to have been adequate and efficient. However, we do not know for certain what we would have gained by using more comprehensive methods and we do not know whether the resulting intervention would have been more effective if we had used other methods. The effectiveness of our multifaceted intervention is under evaluation in a randomised controlled trial. PMID- 15347427 TI - Graft calcifications and dysfunction following liver transplantation. AB - BACKGROUND: The molecular events, following ischemia and reperfusion (I/R) of the liver during transplantation are largely unknown. There is evidence that apoptotic and necrotic events may take place, and occasionally result in primary graft dysfunction. We herein report two cases, where significant I/R injury correlated with the development of liver calcification and primary liver dysfunction. CASE PRESENTATION: Both patients with clinical and biochemical evidence of primary graft dysfunction demonstrated calcification at light and electron microscopy levels. In addition, one patient had macroscopic evidence of calcification on cross-sectional imaging. Both patients died secondary to the sequelae of the graft dysfunction. CONCLUSIONS: Severe I/R-induced injury to the liver, clinically leads to graft dysfunction. This is due to advanced apoptotic and/or necrotic events at the hepatocyte level that may, on the most severe form, lead to calcification. The study of microcalcification at the early posttransplant period could provide insight in the events taking place following significant ischemia/reperfusion-induced injury to the graft. PMID- 15347428 TI - Effects of age and leg length upon central loop of the gastrocnemius-soleus H reflex latency. AB - BACKGROUND: central loop of the gastrocnemius-soleus H-reflex latency (Tc) that looks promising in the diagnosis of S1 radiculopathy; has been investigated in a few studies and only two of them have focused on the constitutional factors affecting it. Although leg length has been shown to contribute to the Tc, the role of age is controversial. More confusing, none of the previously performed studies have used strict criteria to rule out subclinical neuropathy, so the results could be misleading. This study has been performed to determine the influence of leg length and age on Tc among a carefully selected group of healthy volunteers. METHODS: after screening forty six volunteers by taking history, physical examination and a brief electrophysiologic study; forty of them were selected to enroll into the study. Tc was obtained in all the study subjects and leg length and age were recorded for correlational analyses. RESULTS: this group was consisted of 26 males (65%) and 14 females (35%) with the age range of 19-65 years (Mean +/- SD: 37 +/- 10.7) and leg length range of 29.5-43 centimeters (36.4 +/- 3.4). Mean +/- SD for Tc was 6.78 +/- 0.3. We found a significant correlation between Tc and leg length (p value= 0.003, r = 0.49 and confidence interval 95% = 0.59-0.88), no significant correlation was found between age and Tc (p value= 0.48, r = 0.11), also we obtained the regression equation as: Tc = 0.04L + 5.28 CONCLUSIONS: in contrast to leg length, age was not correlated with Tc. Future studies are required to delineate other contributing factors to Tc. PMID- 15347429 TI - Mutational dynamics of the SARS coronavirus in cell culture and human populations isolated in 2003. AB - BACKGROUND: The SARS coronavirus is the etiologic agent for the epidemic of the Severe Acute Respiratory Syndrome. The recent emergence of this new pathogen, the careful tracing of its transmission patterns, and the ability to propagate in culture allows the exploration of the mutational dynamics of the SARS-CoV in human populations. METHODS: We sequenced complete SARS-CoV genomes taken from primary human tissues (SIN3408, SIN3725V, SIN3765V), cultured isolates (SIN848, SIN846, SIN842, SIN845, SIN847, SIN849, SIN850, SIN852, SIN3408L), and five consecutive Vero cell passages (SIN2774_P1, SIN2774_P2, SIN2774_P3, SIN2774_P4, SIN2774_P5) arising from SIN2774 isolate. These represented individual patient samples, serial in vitro passages in cell culture, and paired human and cell culture isolates. Employing a refined mutation filtering scheme and constant mutation rate model, the mutation rates were estimated and the possible date of emergence was calculated. Phylogenetic analysis was used to uncover molecular relationships between the isolates. RESULTS: Close examination of whole genome sequence of 54 SARS-CoV isolates identified before 14th October 2003, including 22 from patients in Singapore, revealed the mutations engendered during human-to Vero and Vero-to-human transmission as well as in multiple Vero cell passages in order to refine our analysis of human-to-human transmission. Though co-infection by different quasipecies in individual tissue samples is observed, the in vitro mutation rate of the SARS-CoV in Vero cell passage is negligible. The in vivo mutation rate, however, is consistent with estimates of other RNA viruses at approximately 5.7 x 10-6 nucleotide substitutions per site per day (0.17 mutations per genome per day), or two mutations per human passage (adjusted R square = 0.4014). Using the immediate Hotel M contact isolates as roots, we observed that the SARS epidemic has generated four major genetic groups that are geographically associated: two Singapore isolates, one Taiwan isolate, and one North China isolate which appears most closely related to the putative SARS-CoV isolated from a palm civet. Non-synonymous mutations are centered in non essential ORFs especially in structural and antigenic genes such as the S and M proteins, but these mutations did not distinguish the geographical groupings. However, no non-synonymous mutations were found in the 3CLpro and the polymerase genes. CONCLUSIONS: Our results show that the SARS-CoV is well adapted to growth in culture and did not appear to undergo specific selection in human populations. We further assessed that the putative origin of the SARS epidemic was in late October 2002 which is consistent with a recent estimate using cases from China. The greater sequence divergence in the structural and antigenic proteins and consistent deletions in the 3'--most portion of the viral genome suggest that certain selection pressures are interacting with the functional nature of these validated and putative ORFs. PMID- 15347430 TI - An economic analysis of premarriage prevention of hepatitis B transmission in Iran. AB - BACKGROUND: To assess the economic aspects of HBV (hepatitis B virus) transmission prevention for premarriage individuals in a country with cultural backgrounds like Iran and intermediate endemicity of HBV infection. METHODS: A cost-effectiveness analysis model was used from the health care system and society perspectives. The effectiveness was defined as the number of chronic HBV infections averted owing to one of the following strategies:1) HBsAg screening to find those would-be couples one of whom is HBsAg positive and putting seronegative subjects on a protection protocol comprising HBV vaccination, single dose HBIG and condom protection.2) HBsAg screening as above, in addition to performing HBcAb screening in the HBsAg negative spouses of the HBsAg positive persons and giving the protocol only to HBcAb negative ones.Sensitivity and threshold analyses were conducted. RESULTS: The cost of each chronic infection averted was 202$ and 197$ for the strategies 1 and 2, respectively. Sensitivity analysis showed that strategy 2 was always slightly cheaper than strategy 1. The discounted threshold value for the lifetime costs of chronic liver disease, above which the model was cost saving was 2818$ in strategy 1 and 2747$ in strategy 2. CONCLUSIONS: Though premarriage prevention of HBV transmission in the countries with cultural backgrounds similar to Iran seems cost saving, further studies determining precise costs of HBV infection in Iran can lead to a better analysis. PMID- 15347431 TI - Enhancer trapping in zebrafish using the Sleeping Beauty transposon. AB - BACKGROUND: Among functional elements of a metazoan gene, enhancers are particularly difficult to find and annotate. Pioneering experiments in Drosophila have demonstrated the value of enhancer "trapping" using an invertebrate to address this functional genomics problem. RESULTS: We modulated a Sleeping Beauty transposon-based transgenesis cassette to establish an enhancer trapping technique for use in a vertebrate model system, zebrafish Danio rerio. We established 9 lines of zebrafish with distinct tissue- or organ-specific GFP expression patterns from 90 founders that produced GFP-expressing progeny. We have molecularly characterized these lines and show that in each line, a specific GFP expression pattern is due to a single transposition event. Many of the insertions are into introns of zebrafish genes predicted in the current genome assembly. We have identified both previously characterized as well as novel expression patterns from this screen. For example, the ET7 line harbors a transposon insertion near the mkp3 locus and expresses GFP in the midbrain hindbrain boundary, forebrain and the ventricle, matching a subset of the known FGF8-dependent mkp3 expression domain. The ET2 line, in contrast, expresses GFP specifically in caudal primary motoneurons due to an insertion into the poly(ADP ribose) glycohydrolase (PARG) locus. This surprising expression pattern was confirmed using in situ hybridization techniques for the endogenous PARG mRNA, indicating the enhancer trap has replicated this unexpected and highly localized PARG expression with good fidelity. Finally, we show that it is possible to excise a Sleeping Beauty transposon from a genomic location in the zebrafish germline. CONCLUSIONS: This genomics tool offers the opportunity for large-scale biological approaches combining both expression and genomic-level sequence analysis using as a template an entire vertebrate genome. PMID- 15347432 TI - Current developments in hormonal therapy of breast cancer. PMID- 15347433 TI - The ATAC (Arimidex, Tamoxifen, Alone or in Combination) trial: an update. AB - Until recently, tamoxifen was the only adjuvant endocrine therapy for postmenopausal women with hormone receptor-positive, early-stage breast cancer. However, the first efficacy update from the ATAC (Arimidex, Tamoxifen, Alone or in Combination) trial has introduced a choice of adjuvant therapy in this setting. After a median follow-up of 47 months, these data indicated that anastrozole continued to show superior efficacy compared with tamoxifen, including significantly greater disease-free survival, a longer median time to recurrence, and a reduced incidence of contralateral breast cancer. Anastrozole also exhibited a number of important tolerability benefits compared with tamoxifen, including reduced incidences of thromboembolic events, vaginal bleeding, and endometrial cancer. Additional ATAC subprotocols included the examination of bone mineral density and its associated sequelae, endometrial abnormalities, and quality of life. The results of these studies support the primary conclusions of the main efficacy and safety analyses: the efficacy benefits of anastrozole were not at the expense of quality of life, anastrozole was associated with reduced endometrial stimulation compared with tamoxifen, and only patients receiving tamoxifen had endometrial atypical hyperplasia. Anastrozole was associated with a modest loss of bone mineral density and an initial increase of fractures compared with tamoxifen. However, the fracture rate with anastrozole stabilizes after 2 years, and indirect comparison suggests that any increased fracture risk with anastrozole is modest. This review concludes that, based on the overall risk-benefit profile from the ATAC study, anastrozole is a rational alternative to tamoxifen for the adjuvant treatment of women with hormone-sensitive early-stage breast cancer. PMID- 15347434 TI - Switching trial of adjuvant tamoxifen with an aromatase inhibitor in postmenopausal patients with breast cancer. AB - Tamoxifen is currently a standard of care for postmenopausal patients with breast cancer with hormone receptor-positive tumors who are candidates for adjuvant endocrine therapy. This treatment is highly effective and relatively safe. However, a significant proportion of women are constitutively resistant or become resistant to tamoxifen, despite having hormone receptor-positive tumors. Moreover, the prolonged exposure to tamoxifen can produce severe and life threatening side effects like endometrial carcinoma or thromboembolic disease. Aromatase inhibitors (AI) have been shown to be quite effective in advanced disease, and also have promising efficacy in early-stage breast cancer as alternatives to tamoxifen. This article reviews the results achieved by AI following 2-3 years or 5 years of tamoxifen. At least 3 trials indicate that switching to an AI following 2-3 years of tamoxifen can produce a substantial benefit. A fourth trial indicates that additional benefit can be achieved by a few years of treatment with an AI after 5 years of tamoxifen. The updated results of previous trials and ongoing trials will soon establish criteria for selecting patients who might be better candidates for sequencing, and to fine tune strategies that are more appropriate. PMID- 15347435 TI - Adjuvant aromatase inhibitors following tamoxifen for early-stage breast cancer in postmenopausal women: what do we really know? AB - Adjuvant hormonal therapy in the treatment of women with early-stage, hormone receptor (HR)-positive breast cancer is now considered the standard of care. Adjuvant tamoxifen decreases the risk of breast cancer recurrence and death in women with early-stage breast cancer when taken for 5 years. The benefits of tamoxifen are counterbalanced by toxicities including an increased risk of endometrial cancer and thromboembolic events. The selective aromatase inhibitors (AIs)--including anastrozole, letrozole, and exemestane--are challenging the role of tamoxifen as the adjuvant hormonal therapy of choice in postmenopausal women. Results of the Arimidex and Tamoxifen Alone or in Combination trial favor the use of anastrozole over tamoxifen as initial adjuvant hormonal therapy, with improvement in disease-free survival (DFS) and a favorable toxicity profile. The results of 2 large adjuvant trials using AIs sequentially with tamoxifen in postmenopausal women with early-stage, HR-positive breast cancer have been reported. The MA-17 study randomized women to placebo or letrozole for 5 years after completion of 4.5-6 years of initial tamoxifen. The Intergroup Exemestane Study (IES) randomized women following 2-3 years of adjuvant tamoxifen to continue to receive tamoxifen or switch to exemestane for a total of 5 years of adjuvant hormonal therapy. The MA-17 and IES trials demonstrated superior DFS with the AI and corroborated the smaller GROCTA-4B and Italian Tamoxifen Arimidex trials, which studied sequential therapy with aminoglutethamide or anastrozole. There is now substantial medical evidence supporting the use of AIs in postmenopausal women with early-stage, HR-positive breast cancer. PMID- 15347436 TI - Implications of first-line adjuvant treatment with aromatase inhibitors in recurrent metastatic breast cancer. AB - The role of adjuvant tamoxifen therapy has gone unchallenged until recently. With the introduction of the selective aromatase inhibitors (AIs), the paradigm for treatment of hormone receptor-positive breast cancer in postmenopausal women is changing. New data from randomized clinical trials have shown the impact of the use of an AI compared with tamoxifen or in sequence with tamoxifen. This review will emphasize some of the highlights from these data sets and the limitations of our current knowledge. Finally, we will discuss the implications of the use of nonselective AIs in the adjuvant setting for the patient who develops recurrent metastatic disease. PMID- 15347437 TI - [History, status quo and expectation of primary immunodeficiency]. PMID- 15347438 TI - [Clinical features of X-linked agammaglobulinemia: analysis of 8 cases]. AB - OBJECTIVE: X-linked agammaglobulinemia (XLA), caused by mutations in Bruton's tyrosine kinase (BTK), is a common form of inherited antibody deficiency. There were very few case reports of this disease that were diagnosed only based on clinical findings in China. The purpose of this study was to evaluate the clinical features of 8 Chinese cases with XLA with BTK defect which were confirmed by flow cytometry and/or gene analysis. METHOD: Based on clinical findings, 8 suspected XLA patients were confirmed by detecting the expression of BTK by flow cytometry and/or gene analysis of BTK. The history and thorough physical examination and routine immunological evaluation of 8 cases were collected and reviewed. RESULTS: The age of onset of all the 8 male patients were from 3 months to 3 years. The mean age at diagnosis was 6 years. Recurrent upper respiratory infection and pneumonia with fever were seen in all the patients. Nasopharynx infection was mainly contributed to upper respiratory infection. Very few or no otitis (1/8) and sinusitis (0/8) were involved. Polyarthritis without evidence of infection was common (3/8). Chronic diarrhea was documented during the first 2 years after the onset of the disease in 2 cases. Two of the patients suffered from meningitis one time each. Skin infection was not serious in two patients. Osteomyelitis occurred in one case, which occurred secondary to a trauma. One case had poliomyelitis-like disease that was considered to be related to polio vaccine. Only two cases had unconfirmed maternal family history of XLA. The prominent signs at diagnosis were dystrophia, growth and developmental retardation and markedly decreased or absent tonsils and lymph nodes. Concentration of all classes of serum immunoglobulins (Igs) and the number of B cells in the peripheral circulation were dramatically decreased. The ratio of CD4/CD8 in most of the patients (6/8) was markedly inverse. CONCLUSION: The age at diagnosis of this reported group was older. Clinical symptoms displayed recurrent upper respiratory infection (nasopharynx infection but rare or no otitis or sinusitis) and pneumonia; polyarthritis was common. There were no confirmed family history of XLA. Most of the patients showed inverse ratios of CD4/CD8, the reason and potential significance are unclear. PMID- 15347439 TI - [Clinical features of primary Sjogren's syndrome in childhood]. AB - OBJECTIVE: To assess the clinical features of primary Sjgren's syndrome (pSS) in childhood and adult patients to understand the differences between them. METHOD: Data of 21 childhood and 400 adult patients with definite primary Sjgren's syndrome were analyzed retrospectively. RESULTS: In adult patients, initial clinical symptoms were various, dry mouth and dry eyes, arthritis and arthralgia, parotid swelling, renal tubular abnormalities (RTA) were more common. Compared with adult patients, RTA, parotid swelling and rash were more common in childhood, pulmonary abnormalities and neurologic system involvement were less common. The frequency of renal tubular abnormalities (52.4%) and rash (47.6%) in childhood pSS were higher than those of adults, but the frequencies of dry eyes (61.0%) and pulmonary interstitial fibrosis (25.2%) in adults pSS were higher than those in childhood (P < 0.01); serum rheumatoid factor (RF) and gamma globulin were higher in all childhood patients (P < 0.01). Anti-nuclear antibody (ANA), anti-SSA and anti-SSB antibodies, and IgG were significantly higher in childhood cases than in adult patients (P < 0.05 or 0.01). CONCLUSION: The major clinical characteristics of childhood pSS cases included: (1) RTA, parotid swelling and rash which appeared earlier, but dry mouth and dry eyes appeared later and were mild. (2) The frequency of pulmonary abnormalities, nervous system involvement and Raynaud's phenomenon were less commonly seen. (3) Positivity of RF, anti-SSA and anti-SSB antibodies, and serum IgG were higher. PMID- 15347440 TI - [Clinical significance of antiphospholipid antibody in pediatric patients and review of literature]. AB - OBJECTIVE: Antiphospholipid antibody (APL) is a particularly important laboratory diagnostic criterion for antiphospholipid syndrome (APS). The significances of positive APL in childhood are seldom reported nor fully understood. The purpose of this study was to analyze 13 cases with positive APL seen in our hospital and to study the relationship between the positive rates of APL and various clinical diseases especially systemic lupus erythematosus (SLE) in order to improve the clinical diagnoses and treatment level of APS in children. METHODS: The clinical data collected from 2000 to 2002 of 13 hospitalized children with positive APL were retrospectively evaluated. Enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence technique were used respectively to detect APL and antineutrophil cytoplasmic autoantibodies (ANCA) of sera from those children. Other various indexes were also detected according to different characteristics of different diseases. RESULTS: Eight cases had SLE; 2 had acute post streptococcal infections. The other 3 cases did not show any evidences of primary diseases; they probably had primary APS. SLE was the most common primary diseases to cause development of APL and the cases with SLE showed more severe cutaneous vasculitis than SLE patients who were negative for APL. There was no significant relationship between the positive rates of APL and that of ANCA. Eight APL positive cases complicated with thrombocytopenia and bleeding were treated with high dosage of immunoglobulin [400 mg/(kg.d), for 3 - 5 d] intravenously; the clinical conditions of these cases were ameliorated soon. While the 5 cases who had thrombotic vasculitis and thromboembolism were treated with anticoagulant and antithrombotic therapy with low molecular weight heparin [50 - 100 U/(kg.d)], which led to good clinical effects. CONCLUSIONS: The clinical manifestations of children positive for APL were somehow different from those of adults. Positive APL itself may be nonspecific, it can occur from different causes of diseases. APL detection may be useful to suggest anticoagulant and/or antithrombosis therapy. Treatments for APS should be variable according to different causes and severity of diseases, in the cases of thrombocytopenia and bleeding, high dose intravenous immunoglobulin should be given as soon as possible, while in the cases of thrombotic vasculitis and thromboembolism, anticoagulant and antithrombotic therapy should be given soon. PMID- 15347441 TI - [Long-term outcome after injection sclerotherapy for esophageal variceal bleeding in children with portal hypertension]. AB - OBJECTIVE: Endoscopic sclerotherapy has emerged as an effective treatment for bleeding esophageal varices in adults and children but the long-term outcome is poorly defined in children. The present study aimed to study the long-term effect of endoscopic sclerotherapy in children with portal hypertension. METHODS: Fifteen patients (age 3 to 14 years) with esophageal variceal bleeding underwent endoscopic injection treatments with 1% Aethoxy-sclerol since 1996. All subjects continued to receive the therapy by repeated intra and extravariceal endoscopic sclerotherapy at intervals of 3 - 4 weeks until the varices disappeared, and received regular endoscopic follow-up. RESULTS: Fifteen patients had totally 43 injections, and were followed up from 40 to 86 months (mean 66 months) by endoscopy. Two patients received 2 injections and 5 received 3 before eradication of varices. The mean time needed for varices eradication was 3 to 6 months. Recurrence of varices and bleeding was seen in 3 patients who had duodenal ulcer. CONCLUSION: Endoscopic sclerotherapy is a safe and effective treatment for pediatric esophageal varices. PMID- 15347442 TI - [Association of HLA-DQA1 loci with Helicobacter pylori infection in children of Guangxi Zhuang, Mulam nationalities, China and central Jing nationality, Vietnam]. AB - OBJECTIVE: To compare the prevalence of Helicobacter pylori (Hp) infection in children of Zhuang and Mulan ethnic groups, Guangxi Luocheng county, China and in children of Jing ethnic group, Central Vietnam and to analyze the association of HLA-DQA1 alleles of these ethnic groups' children with Hp infection. METHODS: Serodiagnosis by determining Hp antibody with ELISA and determination of serum CagA, VacA and urease antibodies by immunoblotting were performed for 54 Zhuang, 76 Mulan and 109 Jing children. Polymerase chain reaction-single strand polymorphism (PCR-SSP) technique was applied to determine the polymorphism of the HLA-DQA1 locus of these children and then the association of HLA-DQA1 alleles of these minority children with Hp infection was analyzed by SAS software. RESULTS: The prevalence of Hp infection were 39% in Vietnamese Jing nationality, which was significantly lower than that in children of Guangxi Luocheng county (65% in Zhuang nationality and 58% in Mulan nationality) (P < 0.01). The distribution of HLA-DQA1 locus was not significantly different among the 3 groups. The frequency of HLA-DQA1 * 0104 allele was significantly higher in children with Hp infection than in children without Hp infection in each of the 3 groups (P < 0.01). CONCLUSION: The results indicated that the prevalence of Hp infection in Zhuang and Mulan minority ethnic groups in Guangxi, China was higher than that in Vietnamese Jing ethnic group children. HLA-DQA1 * 0104 allele may be associated with susceptibility to Hp infection. PMID- 15347443 TI - [Diagnosis and treatment of methylmalonic acidemia in 14 cases]. AB - OBJECTIVE: Methylmalonic acidemia (MMA) is one of the most common disorders of congenital organic acid metabolism. This study aimed at exploring the clinical characteristics and treatment of the disease to help improve our understanding of it. METHODS: The clinical data of 14 patients with MMA admitted to our hospital between January 2002 and July 2003 were analyzed and the diagnoses were confirmed by gas chromatography/mass spectrometry (GC/MS). The patients consisted of 4 males and 10 females, whose age of onset ranged from birth to 9 years with 7 cases younger than 1 month (50%) and 10 cases younger than 1 year (71%). RESULTS: The main clinical manifestations were lethargy (6 cases), developmental retardation or regradation (7 cases), convulsion (6 cases), recurrent vomiting (4 cases), difficulty with feeding (4 cases), muscular dystonia (5 cases with hypotonia, 3 with hypertonia) and yellowish hair (4 cases), etc. Some cases were also presented with hair loss, hepatomegaly, ataxic or stiff gait, and motor weakness with muscular atrophy. The laboratory findings showed metabolic acidosis in 6 cases, hyperammonemia in 5 cases, ketonuria in 4 cases and remarkable elevation of urinary methylmalonic acid concentration in all cases. Some abnormalities in globus pallidus and cerebral white matter as well as diffuse cerebral atrophy were noted by the brain CT and MRI in 5 respective cases, while 4 cases did not receive neuroradiological examinations. Peripheral neuropathies were found by electromyography in 2 patients and bilateral optic nerve atrophy was detected by eyeground examination in 1 child. Three patients died before the diagnoses were made. Of the 11 survivals, 10 children have received therapy of vitamin B12 (VitB12) and supplementation of L-carnitine with restricted-protein diet. The follow-up for a period ranging from 3 months to 1.5 year (mean 8.5 months) of 7 cases with medical therapy showed a favorable outcome without any symptoms in 1 case and apparent improvement in 4 cases (the diffuse cerebral atrophy in MRI completely recovered in one case), however, 2 patients died from severe metabolic acidosis. CONCLUSIONS: The main clinical features of MMA include lethargy, developmental retardation or regradation, convulsion, recurrent vomiting, difficulty with feeding, muscular dystonia, yellowish hair, metabolic acidosis, hyperammonemia and ketonuria, etc. Urine organic acids analysis with GC/MS is critical to the early diagnosis of MMA. Early diagnosis and appropriate long-term treatment are essential to improve the prognosis of the disease. PMID- 15347444 TI - [Molecular genetic analysis of congenital lipoid adrenal hyperplasia]. AB - OBJECTIVE: Congenital lipoid adrenal hyperplasia (CLAH) is an autosomal recessive inherited disorder, characterized by deficiency of adrenal and gonadal steroid hormones. Recent studies have shown that mutations in the gene for steroidogenic acute regulatory protein (StAR) cause this most severe genetic disorder in steroid hormone biosynthesis. StAR is a mitochondrial protein promotes cholesterol transfer from outer mitochondrial membrane to the inner mitochondrial membrane, where the cholesterol serves as a substrate for P450scc and initiates steroidogenesis. So far, more than 30 different mutations in the StAR gene have been found in the patients with CLAH from various ethnic groups. None of CLAH patients in the Chinese population has been previously reported. In the present study we analyzed the StAR gene in a Chinese patient with CLAH. METHODS: The patient who was a 19-yr-old phenotypic female, has a 46, XY karyotype. Endocrinological evaluation was performed. Genomic DNA samples were abstracted from the bloods of the patient and his parents. Polymerase chain reaction (PCR), direct DNA sequencing, family analysis and restriction enzyme digestion analysis were used to detect and confirm the mutations of StAR gene. RESULTS: Endocrine evaluation of the patient showed extremely elevated basal concentrations of serum ACTH and gonadotropin and minimal concentration of gonadal steroids. An ACTH stimulation test indicated basal serum dehydroepiandrosterone and 17 hydroxyprogesterone were lower than normal detectable range and had no obvious increase after the ACTH stimulation. Automatic sequencing of 7 exons of the StAR gene with the polymerase chain reaction products of the genomic DNA revealed compound heterozygous for a novel nonsense mutation Q77X in exon 3 and the frameshift mutation 838delA in exon 6. The father carried Q77X mutation and the mother carried 838delA mutation. The restriction enzyme site of the Q77X mutation was examined by endonucleotidase BfaI. Furthermore, this mutation was not found in a series of 20 alleles of normal individuals. CONCLUSION: Q77X is the novel mutation found in the patient with CLAH. Q77X and 838delA compound mutations could inactivate the StAR function and give rise to clinically manifest CLAH. This case is the first Chinese patient with CLAH identified by molecular genetic analysis. DNA-based analysis of StAR gene will be helpful for the diagnosis of CLAH. PMID- 15347445 TI - [Proteins structure changes of COL4A5 gene point mutations and structure phenotype relations in Alport syndrome]. AB - OBJECTIVE: Alport syndrome (AS) is a progressive hereditary nephritis presented with hematuria and renal failure, frequently associated with sensorineural deafness and ocular lesions. So far, more than 300 gene mutations in AS have been identified which provides a better way to analyze the association between genotype and phenotype. It is hard to understand all the phenotype according to the gene mutations, because the structure and function changes of the relevant protein, alpha5(IV) chain, encoded by mutated COL4A5 gene are rare to know. This study aimed to detect the proteins structure encoded by COL4A5 gene with different missense mutations and to analyze the effect of gene mutations on the secondary structure of alpha5(IV) chain and structure-phenotype relations. METHODS: Two X-linked AS patients with different missense mutations (g.3246G > T resulting in p.G1015V and g.3290G > A resulting in p.G1030S, respectively) diagnosed by clinical manifestations, family history and skin or renal biopsy examinations, as well as a control were included in this study. The fragments of cDNA with the two mutations, respectively, and that of corresponding cDNA from the control were expressed in E. coli. The secondary structure of the recombinant polypeptides were analyzed by using circular dichroism (CD) spectroscopy. RESULTS: CD spectra of the control exhibited a negative peak near 200 nm whereas that of the patient 1 exhibited a negative peak near 220 nm. Furthermore, the magnitude of the negative peak of patient 1 decreased from -9000 deg x cm2 x dmol(-1) to -4000 deg x cm2 x dmol(-1) as compared with that of the control. CD spectra of the patient 2 were slightly changed with the negative peak remaining near 220 nm but the magnitude increasing from -9000 deg x cm2 x dmol(-1) to 11000 deg x cm2 x dmol(-1) as compared with that of the control. In addition, the secondary structure of the control polypeptide was mainly composed of beta-sheet and random coil without alpha-helix, whereas that of the patient 1 presented 12.9% alpha-helix. Although the secondary structure of polypeptide of the patient 2 was also mainly composed of beta-sheet and random coil, the composition of beta sheet reduced and random coil increased. CONCLUSION: Although the glycine substitutions existed in the same domain of alpha5(IV) chain, the patient 1 with the severe AS phenotype and g.3246G > T mutation, and patient 2 with the mild AS phenotype and g.3290G > A mutation were revealed with different secondary structures of alpha5(IV) chain. Moreover, the secondary structure changes of alpha5(IV) chain were consistent with their corresponding phenotype severity. PMID- 15347446 TI - [Comparitive [Comparative] study of the indexes of pulmonary arterial pressure of healthy children at different altitudes by Doppler echocardiography]. AB - OBJECTIVE: Reduced oxygen availability at a high altitude is associated with increased pulmonary arterial pressure (PAP). With the altitude goes up the change of PAP in healthy children is still not clear. The difference of PAP in native Tibetan and Han children at a high altitude are also not clear. Many studies have shown that Doppler echocardiographic estimation of PAP correlates closely to the values obtained with the invasive measurement. Therefore the indexes of PAP in health children living at different altitudes were investigated and the indexes of PAP in Han and Tibetan children at the high altitude were compared by Doppler echocardiography. METHODS: A randomized survey was carried out on the indexes of PAP with Doppler echocardiography (HP-8500 and CAL-163 echocardiographic machine) by two doctors of Qinghai Provincial Women and Children Hospital from June 1998 to July 2002. The survey covering Jiuzhi Qinghai (at 3700 m above the sea level), Xining Qinghai (at 2260 m above the sea level) and Shanghai (at 16 m above the sea level) included a population of 1061 children aged 0 - 14 years. The population of 1061 composed of 218 Han children at Shanghai, 567 Han children at Xining Qinghai and 276 children at Jiuzhi, Qinghai including 118 migrated Han and 148 native Tibetan children. The physical, EKG and chest X-ray examination of each child were carried out to ensure all the subjects were healthy. A pulse oximeter was placed on each child's foot to provide measurements of arterial oxygen saturation (SO2) distal to the ductus arteriosus. The indexes of PAP included right ventricular systolic time interval (RSTI) and mean of pulmonary arterial pressure (mPAP) which was assessed by a multiple regression equation (mPAP=27.79 + 35.42 x PEP/AT-50.85 x AT/ETc). The AT/ETc was that AT/ET was divided by R-R. The RSTI included previous ejection period (PEP), ascending time (AT), ejection time (ET), PEP/AT and AT/ET. All subjects were divided into 7 age groups. The data of PAP indexes were compared among three different altitude groups and age groups. The data of PAP indexes were also compared in Han and Tibetan children living over 3700 m sea level in each age group. RESULTS: With the altitude increase the SO2 of the subjects obviously reduced and the indexes of PAP changed. The SO2 correlated closely with the PEP, AT, PEP/A, AT/ET and mPAP (r = 0.352, 0.144, -0.394, -0.166 and -0.363, respectively; P < 0.01). The AT and AT/ET in the groups of 2260 m and 3700 m were shorter than those in the group of 16 m (P = 0.03-0.000) in each age group. The PEP and PEP/AT in 3700 m group were longer than those in 2260 m and 16 m groups (P=0.006-0.000) in each age group. The mPAP in 3700 m group was higher than that in 2260 m and 16 m groups in each age group (mean [+/-SE] mmHg, 35.23 +/- 8.72 vs 17.99 +/- 8.78 and 15.86 +/- 8.96 aged 0 - 28 d, 32.06 +/- 13.38 vs 20.72 +/- 5.71 and 14.64 +/- 8.19 aged to 6 mo, 31.83 +/- 10.53 vs 20.89 +/- 10.12 and 14.69 +/- 5.89 aged to 1 yr, 27.58 +/- 13.55 vs 19.12 +/- 9.75 and 17.36 +/- 6.71 aged to 3 yr, 24.19 +/ 8.38 vs 19.64 +/- 9.36 and 16.43 +/- 4.68 aged to 6 yr, 23.90 +/- 11.35 vs 16.77 +/- 6.79 and 14.42 +/- 6.50 aged to 10 yr, 23.08 +/- 7.31 vs 18.53 +/- 7.25 and 15.45 +/- 6.12 aged to 14 yr, P=0.000). With the growth of the children the reduction of PAP was remarkable at 3700 m above sea level (F=5.638 P=0.000), the mPAP indexes of the first, second and third age groups were evidently higher than those of the other age groups. The SO2, RSTI and mPAP in the native Tibetan children were not different from those in the migrated Han children (P > 0.05) in each age group. CONCLUSION: The PAP of healthy children at the high altitude was different from that of healthy children at the low altitude. The PAP of the healthy children at 3700 m above sea level was remarkably increased. At 3700 m above sea level the PAP of newborns and infants increased much more compared with that of juvenile. The race may not significantly affect the PAP at the high altitude. The high altitude hypoxic environment might play a major role in the increase of PAP. PMID- 15347447 TI - [Plasma exchange in the treatment of atypical hemolytic-uremic syndrome in 2 children]. PMID- 15347448 TI - [Real-time quantitative study of minimal residual disease in childhood B cell acute lymphoblastic leukemia]. AB - OBJECTIVE: The study was aimed to investigate the feasibility and clinical significance of quantitative detection of minimal residual disease (MRD) in childhood acute lymphoblastic leukemia (ALL) by real-time quantitative polymerase chain reaction (RQ-PCR). METHODS: Clonal IgH gene rearrangements of samples at diagnosis were identified by standard PCR assay with consensus primers. Monoclonal IgH gene rearrangements were analyzed using DNAPLOT software. Upstream primers were designed with the Primer Express software and allele specific oligonucleotide developed complementary to the V-D or D-J junction. Samples at diagnosis were serially diluted to generate the patient specific standard curves. RQ-PCR method was used to quantify the MRD of the follow up samples collected at five time points during chemotherapy. To check the quantity and quality of DNA, the investigators used RQ-PCR analysis for the albumin gene. RESULTS: Totally 16 monoclonal IgH gene rearrangements were identified from 34 patients with B-ALL. The analysis of the 16 monoclonal rearrangements showed that the most frequently used V segment was from V3 family and J segment from J4 and J6. The RQ-PCR sensitivity of 10(-4) to 10(-5) was mostly reached. Non-specific amplification was seen in 6 patients. The number of inserted and deleted nucleotides did not appear to be related to the sensitivity (P > 0.05). The correlation coefficients of all 16 standard curves were excellent (> or = 0.99). The mean slope of the standard curves was -3.4 +/- 0.37 and the mean intercept was 24.3 +/- 2.95. MRD analysis of follow up samples from the 16 patients showed an association between high degree of MRD and relapse. There was no apparent relationship between MRD degree at the end of induction chemotherapy and other high risk factors of ALL (P > 0.05). CONCLUSION: The study showed that the above approach with RQ-PCR was applicable to clinical detection of MRD in childhood ALL. Quantitative and dynamic study of MRD was of prognostic importance. PMID- 15347449 TI - [Myocardial matrix metalloproteinases activities in mice with viral myocarditis and their relationship with cardiac function and myocardial collagen amount]. AB - OBJECTIVE: To investigate the dynamic changes of myocardial matrix metalloproteinases (MMPs) activities in mice with viral myocarditis (VM) and their relationships with cardiac function and myocardial collagen amount and to explore the role of MMPs in the pathologic lesion of VM. METHODS: Sixty-five six week-old male DBA/2 mice were obtained from the Chinese Academy of Medical Sciences. They were divided into two groups randomly. Mice in infected group (n=50) were inoculated intraperitoneally with 0.14 ml of coxsackievirus B3 (CVB3, Nancy strain). Control mice (n=15) were inoculated intraperitoneally with 0.14 ml of Eagle's solution. Eight infected mice were sacrificed on day 3, 7, 10, 21 and 30, respectively and fifteen control mice were killed on day 30 after inoculation. Total protein concentration was determined according to the method of Bradford, while MMPs activities were measured with SDS-PAGE type substrate gels embedded with type I gelatin (zymography). Echocardiographic studies were performed under anesthesia with 3% chloralhydrate intraperitoneally (0.01-0.015 ml/g). Cardiac systolic function indexes, such as peak velocity of aorta (Vp) and flow velocity integral of aorta (Vi) were determined by echocardiography. Histological cross sections of hearts were stained with hematoxylin-eosin and myocardial histopathologic scores were counted under optical microscope. Myocardial collagen amount was measured by determination of hydroxyproline quantification. RESULTS: In virus-infected mice, both MMP-2 and MMP-9 activities were increased significantly compared with those in controls and reached the peak on day 10 (P < 0.01). On day 10, cardiac systolic function indexes (Vp and Vi) were all significantly lower than those at other stages after virus inoculation and in control group (P < 0.05). There was no obvious elevation in myocardial collagen amount in mice with VM at acute stage (P > 0.05). While the myocardial collagen amount in infected group at recovery stage (on day 21 and 30) increased significantly compared with controls. MMP-2 and MMP-9 activities positively correlated with myocardial histopathological scores, respectively (r =0.801, 0.821 P < 0.01), while they negatively correlated with Vp (r = -0.649, -0.683, P < 0.01) and Vi, respectively (r = -0.711, -0.755, P < 0.01). However, Vp and Vi negatively correlated with myocardial histopathological scores (r = -0.756, 0.584, P < 0.01). CONCLUSIONS: In mice with VM, the activities of myocardial MMP 2 and MMP-9 at acute stage increased significantly, then myocardial collagen amount elevated in recovery stage. These changes were associated with myocardial remodeling and cardiac dysfunction. Myocardial MMP activities are important markers of myocardial pathologic lesion. They are of value in the evaluation of the severity of myocardial damage and cardiac dysfunction in mice with VM. PMID- 15347450 TI - [Review and analysis of 283 cases of Kawasaki disease]. AB - OBJECTIVE: The aim of the study was to review the cases of Kawasaki Disease (KD) and analyze the clinical features especially their cardiac complications. METHODS: Totally 283 patients with KD were hospitalized from 1992 to 2002. Their clinical features and factors associated with increased risk of coronary artery aneurysms were reviewed. RESULTS: (1) Among the 283 KD patients, 186 were male and 97 were female. The male-female ratio was 1.9:1. Most of them (71%) were younger than 3 years old. Seasonal peak was in spring and summer (from May to Aug). Depending on the criteria of KD, 228 (81%) were diagnosed as typical KD and 55 (19.4%) were atypical KD. All patients had fever, lasting for 6.1 days. The most common clinical features were oral mucosal changes (97.5%) and cervical lymphadenopathy (95.4%), conjunctivitis (91.2%). And changes in the extremities (89.8%) and rash (81.5%) were also noted. (2) Before the treatment, coronary artery abnormalities were seen in 103/279 (36.9%), which occurred within 4 - 30 days of fever onset. Two weeks after intravenous gamma globulin (IVIG) treatment, the new cases of coronary artery abnormalities were 28/211 (13.3%). The prevalence of coronary artery aneurysms (CAA) with KD was 4.7%. The risk factors of CAA were male cases (P < 0.05) and fever lasting longer than 9 days (P < 0.05). Other cardiac abnormalities in acute phase included left atrial and ventricular enlargement (40/279, 14.3%) and changes in ECG (57/274, 20.8%). The pericardial effusions were found in 11 cases (3.9%). CONCLUSIONS: Cardiac complications of KD occurred in the early period of KD. The new cases of coronary artery abnormalities were 13.3% after IVIG treatment. The risk factors of CAA included male cases and fever lasting for longer time. PMID- 15347451 TI - [Non-dilated coronary artery lesion and myocardial perfusion in children with Kawasaki disease: analysis of 43 cases]. AB - OBJECTIVE: The study was designed to investigate the impact of non-dilated coronary artery wall lesion on myocardial perfusion. METHODS: Doppler tissue image (DTI) was used to measure regional ventricular wall motion in 43 Kawasaki children with non-dilated coronary arterial wall echocardiographic abnormalities (rough intima and arterial wall thickening) detected by two-dimensional echocardiography (2DE) at acute phase. A total of 31 cases who had both non dilated coronary lesion and lowered ventricular wall motion velocity at subacute and convalescence phase underwent submaximal exercise single photon emitting computerized tomography (SPECT) for the evaluation of myocardial perfusion. RESULTS: In 43 cases of Kawasaki disease with non-dilated coronary arterial wall abnormalities, 36 cases (83.7%) still had such lesions at subacute phase and 32 (74.4%)at convalescence. At the same time, lowered regional ventricular wall motion (RVWM) was found in 34 cases at subacute phase and in 31 cases at convalescence. DTI and 2DE had a very good correlation in the detection of such abnormalities (chi(c)2 = 9.64, P < 0.01 in subacute period, and chi(c)2 = 7.14, P < 0.01 in convalescence). In 31 cases accepting SPECT, 17 were positive. A total of 22 ischemic regions were detected. Eighteen out of 22 cases having ischemic regions had abnormal RVWM on DTI. SPECT ischemic regions were significantly in accordance with lowered RVWM in ventricular septum and anterior wall (chi(c)2=5.07 and 7.48, P < 0.05 and P < 0.01, respectively) noted in DTI. CONCLUSION: Non-dilated coronary arterial wall abnormality is one of the forms of coronary artery wall lesions which could reduce myocardial flow perfusion. Its clinical significance is worthy of attention. PMID- 15347452 TI - [One case with pineal germinoma]. PMID- 15347453 TI - [Analysis on causes of death of childhood leukemia from 1981 to 2000 in the city of Tianjin]. AB - OBJECTIVE: Leukemia is a major cause of death of children in China, which accounts for 50 % of all cancers of children. Data from Tianjin Cancer Hospital was analyzed for mortality of leukemia in children under 20 years from 1981 to 2000 in the city of Tianjin. METHODS: All physicians and medical staff of the hospitals and clinics in the registry area were responsible for filling out the report forms for every new case diagnosed as malignant tumors. Death certificates for malignant tumors have been registered at the local police station and the residential files were checked. All cancer cases with insufficient information were traced to his/her family and relevant persons worked in the clinic. Tianjin Cancer Registry Center periodically conducted an active re-checking program to review all patient records on cancers that was not registered in this period. Tumors diagnosed in this study were coded according to the International Classification of Diseases for Oncology (ICD-O). Mortality rates were calculated by age, sex and date of death. RESULTS: The types of acute lymphoid leukemia, acute myeloid leukemia and chronic myeloid leukemia were the most common types of childhood leukemia in Tianjin, comprised 69.3%, 20.9 % and 8.0%, respectively. The mortality for childhood leukemia decreased slowly during the period of 1981 to 2000 in Tianjin. Mortality and morbidity ratios were 0.51. CONCLUSION: Combined with characteristics of individual forms of childhood leukemia mortality, further epidemiological research is needed to prevent childhood leukemia. PMID- 15347454 TI - [Neurogenesis of dentate granule cells following kainic acid induced seizures in immature rats]. AB - OBJECTIVE: Data accumulated over the past years have led to widespread recognition that neurogenesis, the emergence of new neurons, persists in the hippocampal dentate gyrus of the adult mammalian brain, and can be increased by seizures in multiple models. Also, aberrant reorganization of dentate granule cell axons, the mossy fiber sprouting, occurs in human temporal lobe epilepsy and rodent epilepsy models. However a number of studies suggest that the immature brain is less vulnerable to the morphologic alteration of hippocampus after seizures. The goal of this study was to determine whether the seizures can induce dentate granule cell neurogenesis and mossy fiber sprouting in the immature rat. METHODS: Seizures was elicited by unilateral microinfusion of kainic acid (KA, 1 micro g) into the amygdula at postnatal day 15 (P15). Rat pups were given bromodeoxyuridine (BrdU) intraperitoneally on day 5 after KA administration and killed 7 d or 21 d later. The brains were processed for BrdU mitotic labeling combined with double-label immunohistochemistry using neuron-specific, early differentiation marker TuJ1 (betaIII tubulin) or granule-specific marker CaBP (calcium-binding protein calbindin D28k) as well as glia-specific marker GFAP (glial fibrillary acidic protein). Mossy fiber sprouting in intermolecular layer and CA3 subfield was assessed in Timm-stained sections both 1 month and 3 months after KA administration by using a rating scale and density measurement. RESULTS: The dentate BrdU-immunoreactive cells of the KA-treated rats increased significantly compared with those of control rats on day 7 and 21 after BrdU administration (7 d: 244 +/- 15 vs. 190 +/- 10; 21 d: 218 +/- 19 vs. 133 +/- 12, P < 0.05). Approximately 80.2% and 78.7% of BrdU-labeled cells coexpressed TuJ1 in KA-treated rats and control rats on day 7 after BrdU respectively (P > 0.05). On 21 d after BrdU, 60.2% and 58.2% of dentate BrdU-labeled cells coexpressed GaBP in KA-treated rats and control rats respectively (P > 0.05). GFAP colocalized with 3%-5% dentate BrdU-labeled cells in the rats of both groups on day 7 and 21 after BrdU. It was also demonstrated that status epilepticus at P15 did not result in any detectable mossy fiber sprouting within the hippocampus both 1 month and 3 months after KA administration. CONCLUSIONS: KA induced seizures can increase granule cell neurogenesis in the immature rat. Most of newly appeared cells migrate from subgranular proliferation zone (SGZ) into granule cell layer, the hilus as well as the molecular layer, and there they can differentiate into granule neurons. These observations also indicate that there is an early developmental resistance to seizure-induced mossy fiber sprouting in the immature brain. PMID- 15347455 TI - [Evaluation of early and rapid etiological diagnosis of measles by detection of measles virus RNA using nested RT-PCR]. AB - OBJECTIVES: To develop techniques which can be used to detect genetic material of measles virus from clinical samples to make diagnosis and differentiate atypical measles from other exanthematous infections early in clinical course. METHODS: A nested reverse transcriptase-polymerase chain reaction (RT-PCR) was developed to amplify gene fragment with the size of 301 bps from N gene of measles virus in throat swabs and urine samples collected from infants and children who were suspected measles cases. Before the test was used for clinical samples, preliminary tests were performed to determine the sensitivity and specificity of the test. The sensitivity of the test was determined by plaque assay using measles virus strain Edmonton and the specificity of the test was determined by cross-reaction with rubella virus, respiratory syncytial virus, influenza A and B viruses, enterovirus, adenovirus, human cytomegalovirus (hCMV), EB virus, and herpes simplex virus I. Serum specific IgM antibody against measles virus was also tested by ELISA. RESULTS: Measles virus with the titer of 0.53 pfu could be detected by using the nested RT-PCR developed in this study. No amplification was found with the nested RT-PCR when rubella virus, respiratory syncytial virus, influenza A and B virus, enterovirus, adenovirus, hCMV, EB virus, and herpes simplex virus I were used as templates. Out of 116 throat swabs collected from suspected measles cases, 70 (60.3%) were measles RNA positive. For urine samples, 48 out of 74 (64.9%) were positive. Both throat swab and urine samples were collected simultaneously from 73 patients. Among those, 71 (97.3%) showed consistent results. Serum specimens were collected from 110 suspected patients. Among those, 65 (59.1%) and 61 (55.5%) were measles virus specific IgM antibody positive detected with ELISA kits from two different sources, respectively. Out of 110 sera samples, 106 (96.4%) showed consistent results. The consistency of the gene amplification and specific IgM antibody detection was 80.8% as shown by 84 out of 104 patients from whom throat swab and sera were collected at the same time. CONCLUSION: The data indicate that the nested RT-PCR developed in this study is sensitive and specific for detection of gene fragment of measles virus from clinical samples. The test is superior to the commonly used specific IgM antibody detection because of identifying gene material in early clinical stage, and even single clinical sample can be tested. PMID- 15347458 TI - [Summary of the symposium on avian influenza]. PMID- 15347459 TI - [X-linked lymphoproliferative disease: clinical, diagnostic and molecular perspective]. PMID- 15347460 TI - [Eosinophil specific active markers in allergy]. PMID- 15347461 TI - [Evidenced-medicine and its problems in clinical practice]. PMID- 15347462 TI - [Epidemiological analysis of syndactyly in Chinese perinatals]. AB - OBJECTIVE: To study the epidemiological features of syndactyly (SD) in Chinese perinatals. METHODS: Data were collected through Chinese Birth Defects Monitoring Network, a hospital-based congenital malformation registry system. From 1987 through 2001 (except 1994, 1995), all live or still births with 28 weeks of gestation or more born in participating hospitals were assessed within 7 days after delivery. RESULTS: Totally 2311 perinatals with SD were identified among 7 478 746 births, and 57.2% of them were in association with other anomalies. The overall prevalence rate of SD was 3.09/10 000, the rate of isolated SD and associated SD was 1.32/10 000, and 1.77/10 000 respectively. The prevalence rates in urban and rural areas, in male and female births were 3.22/10 000 and 2.79/10 000, 3.42/10 000 and 2.59/10 000 respectively. An increasing trend was found during that period. The perinatal fatality rate of SD was 20.7%, that of isolated form was 5.7%, while that of associated form was 31.9%. The proportion of SD occurring in right side was the same as that in left side, and the proportion of SD in upper limbs equaled to that in lower limbs. CONCLUSIONS: The prevalence rate of SD in Chinese perinatals was similar to that reported in foreign literatures. Associated form of SD was more frequently seen. The prevalence of SD in urban areas was higher than in rural areas. Male excess was identified in both isolated and associated forms of SD. No selective predominance was observed either by affected side or by affected limbs. PMID- 15347463 TI - [Studies on mother-to-infant transmission of hepatitis C virus (HCV) and hepatitis G virus (HGV) co-infection]. AB - OBJECTIVE: To explore mother-to-infant transmission of hepatitis C virus (HCV) and hepatitis G virus (HGV) co-infection and the influence factors. METHODS: Antihepatitis C virus (anti-HCV) and anti-hepatitis G virus (anti-HGV) antibodies were detected by third generation enzyme linked immunosorbent assay (ELISA). HCV RNA and HGV RNA were detected by fluorogenic quantitative-PCR (FQ-PCR). RESULTS: Totally 4506 common pregnant women were tested positive of serum anti-HCV. In these women, 878 were detected of serum anti-HGV, and 10 of them were found with both HCV RNA and HGV RNA positivities. In their 11 infants, two were positive for HCV RNA, and two were positive for HGV RNA. In these 4 infected infants, three were delivered by birth canal, one was delivered by cesarean section. All four were fed by breast-feeding. Three mother's ALTs were abnormally high before delivery. CONCLUSIONS: Hepatitis C and G virus co-infection does not increase the rate of mother-to-infant transmission. Birth canal delivery, breast-feeding and high alanine aminotransferase before delivery are high risk factors for mother-to infant transmission of HCV and HGV co-infection. PMID- 15347464 TI - [Expression of hypoxia inducible factor-1alpha in placenta of pregnancy induced hypertension]. AB - OBJECTIVE: To investigate the expression of hypoxia inducible factor-1alpha (HIF 1alpha) in placentas of pregnancy induced hypertension (PIH). METHODS: Placentas of 49 PIH pregnancies (PIH group 1) and 26 normotensive pregnancies (control group 1) were investigated for HIF-1alpha protein expression using microarray and immunohistochemistry. The expression of HIF-1alpha mRNA in placentas of 12 PIH pregnancies (PIH group 2) and 12 normal pregnancies(control group 2) was respectively detected by RT-PCR. RESULTS: (1) There was significant difference in the positive and slight-moderate-positive spot proportions of HIF-1alpha protein between the PIH group 1 (63.1%, 23.3%) and the control group 1 (20.9%, 7.0%, P < 0.01). (2) The level of HIF-1alpha mRNA between PIH group 2 and control group 2 (0.96 +/- 0.37 vs 0.95 +/- 0.20, P > 0.05) was not statistically different. CONCLUSIONS: (1) There was no significant difference in HIF-1alpha mRNA level between the PIH and normal placentas, but there was a remarkable difference in protein level between the two groups. (2) HIF-1alpha regulate the pathological and physiological progress of PIH at protein level rather than at transcription level. PMID- 15347465 TI - [Study on relationship between serum cholylglycine and placental apoptosis in patients with intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To explore the possible mechanism of placental dysfunction in patients with intrahepatic cholestasis of pregnancy (ICP). METHODS: Serum level of cholylglycine (CG) in 30 cases with ICP (ICP group) and 27 normal pregnant women (control group) was examined by radio-immunoassay before delivery. bax and bcl-2 level in placenta was detected by immunohistochemistry method. RESULTS: (1) Serum CG level in ICP group was (51.8 +/- 5.9) micro mol/L, and in control group it was (9.4 +/- 5.6) micro mol/L. The difference between the two groups was significant (P < 0.05). (2) bax expression level in syncytiotrophoblast of ICP group was: 2( ), 9(+), 11 (++), 8(+++), and bcl-2 expression level was: 12(+), 12(++), 6(+++); bax expression level in control group was: 9 (-), 13(+), 4(++), 1(+++) and bcl-2 expression level was: 3(+), 4(++), 20(+++). bax expression level in syncytiotrophoblast of ICP group was higher than that of the control (P < 0.0005), and bcl-2 expression level was lower than in the control (P < 0.0005), both significantly. A significant positive correlation between CG level and bax expression level and a negative correlation between CG and bcl-2 were found in ICP group (P < 0.005, P < 0.005). CONCLUSION: One of the possible mechanisms involved in placental dysfunction of ICP is overexpression of bax in syncytiotrophoblast caused by high concentrations of bile acid, leading to increased apoptosis. PMID- 15347466 TI - [Dynamic changes of enzymes activities and growth factors contents in human colostrum]. AB - OBJECTIVE: To study the dynamic changes of the activities of superoxide dismutase (SOD), amylase (AM) and alkaline phosphatase (ALP) and of the contents of epithelium growth factors (EGF) and insulin growth factors (IGF-1) in human colostrum (lactation 1 - 7 days postpartum). METHODS: A total of 118 samples of human colostrum were taken from 20 healthy women postpartum. The activities of SOD, AM and ALP in human colostrum were determined by methods of pyrogallol autoxidation, colorimetry of iodine-starch and pyrocatechol phosphate respectively. EGF and IGF-1 contents in human colostrum were measured by radioimmunoassay. RESULTS: The SOD activity of human colostrum increased slightly during lactation 1 - 4 days postpartum, from (18.7 +/- 2.2) kU/L to (22.5 +/- 2.9) kU/L, then it tended to decrease, being (11.2 +/- 2.1) kU/L on the 7th day. The SOD activities in lactation 1 - 5 days postpartum were significantly higher than that on day 7 (P < 0.05 or P < 0.01). AM and ALP activities of human colostrum gradually decreased within a week postpartum. AM activity declined from (155 +/- 12) kU/L to (17 +/- 4) kU/L. ALP activity declined from (105 +/- 17) kU/L to (54 +/- 9) kU/L. AM activities in lactation 1 - 5 days postpartum were significantly higher than that on day 7 (P < 0.05 or P < 0.01); and ALP activity on the first lactation day was significantly higher than that on day 7 (P < 0.05). EGF and IGF-1 concentrations in human colostrum quickly declined during lactation 1 - 4 days postpartum. EGF concentration decreased from (105 +/- 11) micro g/L to (40 +/- 5) micro g/L, and IGF-1 concentration decreased from (25.9 +/- 2.7) micro g/L to (5.6 +/- 1.3) micro g/L. Both concentrations altered slightly during lactation 4 - 7 days postpartum. EGF concentrations in lactation 1 - 3 days postpartum were remarkably higher than that on day 7 (P < 0.05 or P < 0.01); IGF-1 concentration on the first lactation day was significantly higher than that on day 7 (P < 0.01). CONCLUSIONS: Human colostrum contains SOD, AM and ALP with high activities, and is also rich in EGF and IGF-1. All of these are indispensable to normal growth and development of infants. Breast-feeding should be emphasized and encouraged for infants. PMID- 15347467 TI - [Preimplantation genetic diagnosis of chromosome abnormality by fluorescence in situ hybridization]. AB - OBJECTIVE: To perform preimplantation genetic diagnosis (PGD) of chromosome abnormality using fluorescence in-situ hybridization (FISH). METHODS: Ten couples were presented for preimplantation genetic diagnosis. They had a total of 10 oocyte pick-up cycles. The collected oocytes were inseminated by intracytoplasmic sperm injection. PGD was carried out using cleavage-stage (day 3) embryo biopsy, fluorescence in-situ hybridization, and day 4 embryo transfer. RESULTS: Ten oocyte pick-up cycles yielded 158 oocytes. Among the 94 embryos obtained, 54 embryos were biopsied and FISH analyses were performed for 51 blastomeres. Twenty four embryos were transferred on the fourth day. There were 4 clinical pregnancies: 3 infants have been born, and 1 couple had ectopic pregnancy. CONCLUSION: PGD is a valuable method to prevent the high risk of spontaneous miscarriages and conceiving chromosomally unbalanced offspring. PMID- 15347468 TI - [Expression of cystic fibrosis transmembrane conductance regulator in human endometrium]. AB - OBJECTIVE: To study expression of cystic fibrosis transmembrane conductance regulator (CFTR) in human endometrium. METHODS: The expression of CFTR mRNA and protein from 50 samples of normal cyclic human endometrium was examined by in situ hybridization, immunohistochemistry and Western blotting respectively. RESULTS: CFTR mRNA and protein expressions were only detected in the endometrial glandular cells with cyclic changes. CFTR mRNA could be detected from the midproliferative phase with the highest level found in the late proliferative phase, significantly higher than those of late-secretory phase endometrium (P < 0.05). While a large quantity of CFTR protein were seen in late proliferative phase and still presented in the secretive and menstrual phases. Western blotting analysis demonstrated that human endometrium expressed the special CFTR band at 170 000. CONCLUSION: The expression of CFTR in human endometrium presented in glandular cells and the amounts were varied cyclically. The abundant CFTR mRNAs and proteins around the ovulatory period may drive Na(+) and fluid from plasma into the uterine lumen to make optimal electrolyte composition and sufficient fluid volume for sperm migration. PMID- 15347469 TI - [Comparison of the effect of fluoxetine combined with hormone replacement therapy (HRT) and single HRT in treating menopausal depression]. AB - OBJECTIVE: To compare the treatment effects between fluoxetine plus hormone replacement therapy (HRT) and HRT alone on menopausal depression. METHODS: A total of 54 women with climacteric symptoms and depression were enrolled and randomly divided into two groups. The fluoxetine plus HRT group received fluoxetine (20 mg, qd, po) and cyclic use of conjugated estrogen 0.625 mg and medroxyprogesterone acetate 5 mg, while HRT group was assigned to receive cyclic use of conjugated estrogen 0.625 mg and medroxyprogesterone acetate 5 mg only. All subjects were interviewed and evaluated with Hamilton Depression score (HAMD) and Kupperman menopause index (KMI) at week 0, 1, 2, 3, 4, 6 and 8 of the treatment. RESULTS: There was no difference between two groups in HAMD and KMI before the trial (P > 0.05). HAMD scores at the 8th week in fluoxetine plus HRT group and HRT group were 3.0 +/- 2.3 and 11.2 +/- 5.8 respectively, being significantly different (P < 0.001). At the 6th week of treatment KMIs in fluoxetine plus HRT group and HRT group were 9 +/- 6 and 14 +/- 7 respectively, also significantly different (P < 0.05). The healing rates of fluoxetine plus HRT group and HRT group were 92% and 48% respectively, which were statistically different by Chi square test (P < 0.001). CONCLUSIONS: The effect of fluoxetine plus HRT on menopausal depression is significantly superior than that of HRT only and the difference becomes more obvious with treatment time. Both groups could ameliorate the climacteric symptoms with the effect of fluoxetine plus HRT more obvious. PMID- 15347470 TI - [Effect of antisense oligodeoxynucleotide of small subunit component of human ribonucleotide reductase on human choriocarcinoma cell line in vitro]. AB - OBJECTIVE: To study effect of the antisense oligodeoxynucleotide (ASODN) of small subunit of human ribonucleotide reductase (RRM2) mRNA on cell line of human choriocarcinoma in vitro. METHODS: Two 20-mer gapmer ASODNs with a full phosphorothioate backbone were artificially synthesized, which were complementary to nucleotides 626-645 (a coding region) and 1572-1591 (a 3'untranslated region) of RRM2, respectively. ASODNs were transfected into JAR cells through oligofectamine. The survival rate was assessed by methyl thiazolyl tetrazolium (MMT) assay, and RRM2 expression was detected by immunoblot and reverse transcriptase polymerase chain reaction (RT-PCR) methods. RESULTS: Antisense oligodeoxynucleotide one (ASODN1) targeting the coding region significantly inhibited growth of JAR cells in a dose- and time-dependent manner and downregulated RRM2 expression in a time-dependent manner. ASODN1 at 100 nmol/L could inhibit significantly cell growth (P = 0.000), and the effects of ASODN1 on JAR cell proliferation were enhanced with increase of ASODN1 concentration and reached the peak point at 400 nmol/L concentration (P = 0.000). Cell growth was significantly inhibited by 200 nmol/L of ASODN1 after 24 h of treatment (P = 0.000). The effect of ASODN1 was at the maximum at 48 h (P = 0.000), and began to decrease at 72 h of treatment. RRM2 expression started to reduce after ASOND1 treatment for 12 hours, and was obviously downregulated at 24 h of treatment, and decreased to the lowest level at 48 h (P < 0.05). RRM2 expression began to recover at 72 h of treatment. Antisense oligodeoxynucleotide two (ASODN2) corresponding to 3'untranslated region and control oligodeoxynucleotides had no significant effect on the proliferation and RRM2 expression, but the combination of ASODN2 and ASODN1 was more effective on reducing RRM2 expression and inhibiting cell proliferation than ASODN1 alone. CONCLUSIONS: ASODN1 of RRM2 can inhibit the proliferation of JAR cells and downregulate RRM2 transcription and translation in a selective and specific manner. The combination of multiple ASODNs targeting different regions of RRM2 mRNA would be an important approach to improve antisense effectiveness. PMID- 15347471 TI - [Activation of phosphatidylinositol 3-kinase-protein kinase B (PI3K-PKB) induced by 17beta-estradiol in endometrial carcinoma cell (Ishikawa)]. AB - OBJECTIVE: Cellular response to estradiol is mediated both by estrogen receptor (ER) binding to estrogen response element (ERE) and by non-nuclear actions like activation of signal transduction pathways such as mitogen activated protein kinase (MAPK) pathway. However, the signal transduction of estrogen involving phosphatidylinositol 3-kinase-protein kinase B (PI3K-PKB) is not clear in endometrial carcinoma. Our purpose was to study if PI3K-PKB signaling pathway could be activated rapidly by 17beta-E(2) through non-nuclear action and also, whether PI3K inhibitor, LY294002, could inhibit such non-nuclear action of 17beta E(2) in endometrial carcinoma cell line Ishikawa. METHODS: Levels of phosphorylated PKB (Ser473 site, p-PKB) and total PKB were examined by western blotting in Ishikawa cells after stimulation with 17beta-E(2) at 1 x 10(-6) mol/L for different time periods and at varied doses for 30 min. Optimal time and appropriate dose for 17beta-E(2) to activate PKB in Ishikawa cells were observed. Inhibitory effect of LY294002 on activation of PKB induced by 17beta-E(2) was also studied. p-PKB/PKB ratio was used to indicate levels of activation of PKB. RESULTS: p-PKB/PKB at 15 min (0.533 +/- 0.029) was significantly higher than the control (0.361 +/- 0.029, P < 0.05). Maximal activation of PKB (maximal levels of p-PKB/PKB) took place at 30 min (0.666 +/- 0.021, P < 0.001) and the activation persisted at least 2 h after stimulation with 1 x 10(-6) mol/L 17beta-E(2) in Ishikawa cells. With increased doses of 17beta-E(2) (vehicle, 1 x 10(-10), 1 x 10(-8), 1 x 10(-6), 1 x 10(-4) mol/L), the activation of PKB (p-PKB/PKB was 0.300 +/- 0.098, 0.312 +/- 0.081, 0.625 +/- 0.100, 0.771 +/- 0.041, 0.902 +/- 0.043 accordingly) increased gradually (in comparison with vehicle, P > 0.05, < 0.05, < 0.05, < 0.001 accordingly). PI3K inhibitor, LY294002 could inhibit the activation of PKB induced by 17beta-E(2) in Ishikawa cells. Levels of PKB (p-PKB/PKB) decreased (0.443 +/- 0.032, 0.415 +/- 0.032, 0.111 +/- 0.035, 0, 0) gradually with increased concentrations (vehicle, 0.1, 10, 50, 100 micro mol/L) of LY294002 (compared with vehicle, P > 0.05, < 0.05, < 0.001, < 0.001) and 50 micro mol/L LY294002 completely blocked the activation of PKB induced by 17beta-E(2). CONCLUSIONS: 17beta-estradiol, by non-nuclear action, can activate promptly PI3K PKB signaling pathway in endometrial carcinoma cell line, Ishikawa. This action of 17beta-E(2) is PI3K dependent and can be inhibited or blocked by LY294002. PMID- 15347472 TI - [Inhibition of hypoxia-inducible factor 1alpha expression and tumor growth in SKOV3 ovarian cancer model by sirolimus]. AB - OBJECTIVE: To investigate the inhibitory effect of the mammalian target of rapamycin (mTOR) inhibitor, sirolimus on expression of hypoxia-inducible factor (HIF)1alpha protein and growth of ovarian carcinoma in an athymic mouse xenogeneic transplant model of ovarian cancer. METHODS: Four groups of female nude mice were inoculated subcutaneously with SKOV3 cells. After inoculation, mice were treated with saline, rapamycin alone, paclitaxel alone and sirolimus + paclitaxel. In each tumor protein expressions of HIF-1alpha, bcl-2 and apoptosis were determined by immunohistochemistry and RT-PCR. RESULTS: In sirolimus and sirolimus + paclitaxel groups protein expression of HIF-1alpha was inhibited. Tumor burden in rapamycin alone, sirolimus + paclitaxel, and paclitaxel alone was reduced by 47.9% (P < 0.05), 51.0% (P < 0.05), and 31.8% (P > 0.05) respectively compared with controls. Cell apoptosis inder in sirolimus alone (36), sirolimus + paclitaxel (40), paclitaxel alone (22), increased compared with control (15), while expression of bcl-2 decreased compared with control. CONCLUSION: Sirolimus inhibited protein expression of HIF-1alpha, increased tumor apoptosis and decreased tumor growth. PMID- 15347473 TI - [Killing effect of adenovirus mediated fusion gene cytosine and deaminase uracil phosphoribosyl transferase directed by glutathione S-transferase P1 promoter on cisplatin-resistant ovarian cancer cells in vitro]. AB - OBJECTIVE: To construct an adenoviral vector in which the fusion gene cytosine and uracil phosphoribosyl (UPP) transferase was directed by glutathione S transferase P1 (GSTP1) promoter, and to investigate specific killing effect of the suicide gene system on cisplatin-resistant ovarian cancer cells. METHODS: Recombinant adenovirus was generated through homologous recombination in bacteria. A2780 and A2780/DDP cells were infected with Ad and then received flucytosine (5-FC) administration. The relative survival of these cells was tested. And a bystander effect was observed by mixing gene-transferred and gene untransferred A2780/DDP cells with 5-FC. RESULTS: In vitro, when MOI was 100 and 5-FC was 250 micro g/ml, relative survival rate of A2780/DDP cells was only (3.6 +/- 1.0)%; that of A2780 cells was (76.5 +/- 2.8)%. Significant bystander effect was caused by CD-UPP gene and 20% gene-transferred A2780/DDP cells induced 80.3% of total cells to death. CONCLUSION: Recombinant adenovirus carrying CD-UPP gene driven by GSTP1 promoter has a specific killing effect on cisplatin-resistant ovarian cancer cells. PMID- 15347475 TI - [Immortalization of human embryonic cervical epithelial cells induced by E6, E7 genes of human papillomavirus 16]. AB - OBJECTIVE: To establish an immortalized cell line derived from the embryonic cervical epithelium by infection with the recombinant adeno-associated virus (rAAV) containing human papillomavirus (HPV)16 E6, E7, and to study the biological features of cervical cancer cell line. METHODS: Human embryonic cervical tissues were cultured in keratinocyte free serum (K-FS) medium and infected with rAAV containing HPV16 E6, E7. Morphological features and growth rate were examined by light, electronic and fluorescence microscopies. The fragments of E6, E7 were detected by polymerase chain reaction (PCR) and laser confocal microscopy. The biological characteristics of human cervical epithelium were observed by soft agar culture, scid mice inoculation and chromosome analysis. Cell proliferative dynamics was plotted by flow cytometry. RESULTS: After a long-term culture, the phenotype kept the characteristics of primary epithelial cells. They showed monolayer, anchorage-dependent and attachment inhibited growth without forming colonies in soft agar culture. They were non oncogenic when inoculated into scid mice. The tonofilament expression in the cervical cancer cells was inspected by electronic microscopy, demonstrating that the cells were squamous epithelium in origin. The cell line contained HPV16 E6, E7 genes by PCR and laser confocal detection. Chromosome analysis disclosed that the karyotype was diploid or polyploid. The 11th chromosome was assumed to be the integration site by rAAV containing HPV16 E6, E7. CONCLUSIONS: Establishment of the immortalized cervical epithelial cell line by infection with rAAV containing HPV16 E6, E7, supports that HPV16 E6, E7 may be the primary etiology of cervical cancer. It will facilitate further research on the etiology and pathogenesis of cervical cancer. PMID- 15347474 TI - [Alteration of survivin gene expression in ovarian cancer cell line CAOV3 after chemotherapy in vitro]. AB - OBJECTIVE: To study the alteration of survivin gene expression in ovarian cancer cell CAOV3 after chemotherapy in vitro. METHODS: After the secondary culture of ovarian cancer cell line CAOV3, inhibition of ovarian cancer cell growth induced by paclitaxel or carboplatin at different concentration was detected by methyl thiazolyl tetrazolium (MTT). Alteration of survivin mRNA expression was evaluated by reverse transcription polymerase chain reaction (RT-PCR) in ovarian cancer cells which had been cultured with paclitaxel or carboplatin of high concentration (200, 500 mg/L) for 1 d or of low concentration (20, 50 mg/L) for 1, 3 and 5 d, respectively. Meanwhile, CAOV3 cell apoptosis was detected by flow cytometry. RESULTS: The inhibition rates on growth of ovarian cancer cell CAOV3 by different concentrations of paclitaxel or carboplatin after 48 h were 94%, 88%, 34%, 22%, 13% and 97%, 46%, 14%, 9%, 7%. The apoptosis rates of CAOV3 in low concentration paclitaxel for 1, 3, and 5 d were 15%, 23% and 29%. And the apoptosis rate in high concentration paclitaxel for 1 d was 43%. The apoptosis rate of CAOV3 in low concentration carboplatin for 1, 3, and 5 d were 14%, 22% and 26%. And in high concentration carboplatin for 1 d the rate was 19%. Expression of survivin mRNA increased distinctly after culturing in paclitaxel or carboplatin of low concentration for 3 d and of high concentration for 1 d compared to the control group (P < 0.01), the increase was especially evident in paclitaxel treated group (P < 0.01). CONCLUSION: Survivin gene may play an inhibitory role in the chemotherapy of ovarian cancer cell CAOV3 in vitro. PMID- 15347476 TI - [Echocardiography in the prediction of prognosis of irregular fetal heart rhythm]. PMID- 15347478 TI - [Relation between distribution of small heterodimer partner gene and birth weight ]. PMID- 15347479 TI - [Analysis of chromosome examination in 780 cases]. PMID- 15347480 TI - [Ultra-speed cryopreservation of ovary]. PMID- 15347481 TI - [Human papillomavirus infection of reproductive system and local immunity of cervix]. PMID- 15347482 TI - [Progress on the study of ovarian carcinoma vaccine ]. PMID- 15347483 TI - Implementation of dermato-epidemiology curriculum at Case Western Reserve University dermatology program. AB - Dermato-epidemiology curriculum has been identified by the American Academy of Dermatology (AAD) as an important foundation for dermatology residency training. However, no one has yet reported implementation of dermato-epidemiology curriculum. To evaluate and relate our experience carrying out a dermato epidemiology resident education initiative, based on recommendations by the AAD Epidemiology Committee. Monthly lectures based on topics suggested by the AAD Epidemiology Curriculum. Pre- and post-test multiple choice and free-form question measures were employed to examine performance, assess resident enthusiasm, and solicit feedback from the initiative. Quantitative achievement on multiple-choice items improved slightly, but insignificantly, from 53 percent to 58 percent. Resident level of enthusiasm and perceived efficacy for the intervention varied from 3.3 to 4.0 on a 5-point Likert scale where "1" indicates strongly disagree and "5" indicates strongly agree with measures of effectiveness. Dermato-epidemiology curriculum is desirable and achievable even in dermatology programs without full-time epidemiologists. A successful epidemiology curriculum should be clinically and board-examination relevant, incorporating aspects of problem-based, interactive learning. PMID- 15347484 TI - Tacrolimus ointment: utilization patterns in children under age 2 years. AB - Atopic dermatitis (AD) is a common eczematous skin condition; as many as 10-17 percent of all children are affected, and 35-60 percent of affected patients manifest symptoms manifest during the first year of life. Treatment principles for AD in young children involve conservative measures such as avoidance of hot water and environmental irritants, combined with liberal use of emollients after bathing. Low potency topical corticosteroids (TCS) are the current standard of therapy for AD in young children, reserving mid- and high-potency TCS for severe disease. However, complications of long-term use of TCS include skin atrophy, stria formation, telangiectasia, hypopigmentation, secondary infections, steroid acne, allergic contact dermatitis, and miliaria. The pediatric population is also at increased risk for systemic absorption because of their high ratio of skin surface to body mass. Systemic absorption may result in hypothalamic-pituitary adrenal axis suppression and ultimately growth retardation. Although most topical and systemic corticosteroids are not approved by the Food and Drug Administration for use in children less than 2 years of age, conservative treatment often fails in this age group and frequently patients are treated with TCS, antibiotics, and antihistamines. PMID- 15347485 TI - The use of topical tacrolimus and pimecrolimus to treat psoriasis: a review. AB - Tacrolimus ointment and pimecrolimus cream are approved in the United States for treatment of atopic dermatitis. Tacrolimus and pimecrolimus are both calcineurin inhibitors and function as immunosuppressants. Their mechanisms have been discussed elsewhere. This article will discuss their utility in treating psoriasis. PMID- 15347486 TI - Imiquimod 5 percent cream and the treatment of cutaneous malignancy. AB - Imiquimod is a novel synthetic compound that is a member of the imidazoquinolone family of drugs. This class of drugs, which also includes a more potent member, resimiquimod (R-848), is unique having the properties of topical immune response modifiers and stimulators. Imiquimod resembles a nucleoside analogue and is known for its potent induction of endogenous antiviral pro-inflammatory mediators. This article is a review of the relevant literature as it relates to the off-label applications of imiquimod 5 percent cream for treatment of cutaneous preneoplastic and neoplastic conditions including its recently approved indication for treating actinic keratoses. PMID- 15347487 TI - Topical phenytoin for wound healing. AB - Oral phenytoin is used widely for the treatment of convulsive disorders and about half the patients treated develop gingival overgrowth as a side effect. The apparent stimulatory effect has prompted its assessment in wound healing. Studies have shown topical phenytoin to promote healing of decubitus ulcers, venous stasis ulcers, diabetic ulcers, traumatic wounds, burns, and leprosy trophic ulcers. The mechanism of action has been postulated to be multifactorial. The present literature indicates that topical phenytoin deserves further investigation as a wound-healing agent in controlled dose-finding clinical trials. PMID- 15347488 TI - A case of granulomatous rosacea: sorting granulomatous rosacea from other granulomatous diseases that affect the face. AB - Granulomatous rosacea is a variant of rosacea that may present similar to other granulomatous diseases. We present the case of a 45-year-old woman with a 2-year history of facial erythema with multiple papules and pustules on the cheeks, chin, and glabella. The patient responded to minocycline, resulting in healing 6 months without residual scarring. This patient's clinical and histological presentation and treatment outcome are to our assessment consistent with granulomatous rosacea. However, other clinically and histologically related entities will be discussed. These entities include, but are not limited to, perioral dermatitis, granulomatous periorificial dermatitis, lupus miliaris disseminatus faciei, facial afro-caribbean eruption syndrome, and sarcoidosis. PMID- 15347489 TI - Streptococcal toxic shock syndrome and sepsis manifesting in a patient with chronic rheumatoid arthritis. AB - Streptococcal-toxic-shock syndrome is caused by virulent strains of exotoxin producing streptococcus, almost always group-A organisms such as Streptococcus pyogenes. It has often been described in the setting of surgical wounds, burns, childbirth, diabetics, elderly, neonates, and immunocompromised hosts, where the portal of entry is the skin. Our patient was on steroids and nonsteroidal anti inflammatory drugs for chronic rheumatoid arthritis and developed this deadly infection after a fall. PMID- 15347490 TI - A 71-year-old man with spindle-cell neoplasm of unknown origin: a difficult-to diagnose clear-cell sarcoma. AB - A patient initially presented in 1988 with a solitary axillary mass, diagnosed as a high-grade neuroendocrine spindle-cell neoplasm; there was no history of a primary cutaneous malignancy. After subsequent development of a pulmonary nodule in 2001 (14-years post initial diagnosis), the case was reviewed and the possibility of metastatic melanoma was raised. The histopathologic and immunohistochemical profile of this melanocytic neoplasm was diagnostic of clear cell sarcoma (CCS) of tendons and aponeuroses, although the differential diagnosis included malignant melanoma, follicular dendritic and interdigitating cell tumors, malignant peripheral nerve sheath tumor, and a category of so-called PEComas. It is the role of pathologists, particularly dermatopathologists, to distinguish CCS from malignant melanoma, and to alert the clinician, because proper diagnosis ultimately influences treatment. We discuss the immunophenotype, differential diagnosis, and molecular signatures of these neoplasms, and review the pertinent literature on these entities. PMID- 15347491 TI - Acquired lymphangiectasis after breast conservation treatment for breast cancer: report of a case. AB - Acquired lymphangiectasis is a dilatation of lymphatic vessels that can result as a complication of surgical intervention and radiation therapy for malignancy. Acquired lymphangiectasis shares clinical and histological features with the congenital lesion, lymphangioma circumscriptum. Diagnosis and treatment of these vesiculobullous lesions is important because they may be associated with pain, chronic drainage, and cellulitis. We describe patient who had these lesions after treatment for cancer. Although a number of treatment options are available, we have found cryosurgery and electrocautery to be particularly effective. PMID- 15347492 TI - Persistent cutaneous manifestations of hyperoxaluria after combined hepatorenal transplantation. AB - A 30-year-old woman with primary hyperoxaluria type I (PHI) developed livedo reticularis with overlying ulcerations on her legs 16 months after receiving a liver-kidney transplant. A skin biopsy of the lesion showed deposits of calcium oxalate. To our knowledge, there have been no reported cases of livedo reticularis in patients with PH1 after a combined liver-kidney transplant. PMID- 15347493 TI - Asymptomatic annular pink papules on the trunk. PMID- 15347494 TI - Colocalized nevus depigmentosus and lentigines with underlying breast hypoplasia: a case of reverse mutation? AB - Nevus depigmentosus (ND) is classically defined as a congenital nonprogressive hypopigmented macule, stable in size and distribution. There have been many reports of colocalization of ND and lentigines. We describe development of multiple lentigines over ND in a 9-year-old girl along with hypoplasia of the underlying breast. The case is being reported to highlight the phenotypic manifestation of reverse mutation and the coincidental breast hypoplasia that has not been reported before. PMID- 15347495 TI - Papillon-Lefevre syndrome: case report and review of the literature. AB - A 15-year-old boy presented with symmetric, well-demarcated, yellowish, keratotic plaques over the skin of his palms and soles extending onto the dorsal surfaces. Well-circumscribed, psoriasiform, erythematous, scaly plaques were also present on the elbows and knees bilaterally along with dystrophy an transverse grooving of the nails. He also had swollen and friable gums since the age of 3 with subsequent loss of most of his permanent dentition. These findings are consistent with Papillon-Lefevre syndrome. The clinical presentation, differential diagnosis, complications and management of this syndrome are discussed. PMID- 15347496 TI - Papillary cystadenoma of the minor salivary gland of the lower lip. AB - A case report of the papillary cystadenoma from minor salivary gland in lower lip of a 54-year-old man is described. PMID- 15347497 TI - Subungual exostosis of the thumb. AB - A 19-year-old girl presented with painful raised skin lesions over the right thumb for the past one year that had ulcerated over the last one month. On examination, a lobulated nodule of bony-hard consistency, measuring around 2 cm was present over the lateral aspect of the tip of the right thumb. The lesion was tender and the overlying skin showed superficial erosions. Roentgenogram of the hand showed mature bone projecting from the distal end of the terminal phalanx of the right thumb forming a "Y"-like bifurcation. Histology from a bit of excised tissue from the lesion (thumb) showed evidence of mature trabecular bone with a fibrocartilaginous cap in the deep dermis. There was no evidence of malignant change on histology. Local excision of the entire lesion was done and there has been no recurrence till date. PMID- 15347498 TI - Indolent herpetic whitlow of the toe in an elderly patient with diabetic neuropathy. AB - We report a case of indolent herpetic whitlow of the toe occurring in an elderly male patient with poorly controlled diabetes mellitus. In this case, the mechanism of transmission was not clear, although he was in a habit of taking a hot spring bath. This patient's symptoms were unusual for herpes simplex; he had no pain in the presence of diabetic neuropathy. The standard therapeutic dose of acyclovir was not effective in suppressing the lesions, and a higher dose was required to induce complete healing. PMID- 15347499 TI - Leprous osteitis presenting as bone cyst and erosions. AB - A 30-year-old man presented to the Hansen outpatient department with swelling and ulceration of toes for 2 months and swelling of the right fifth and fourth fingers and the left second finger for 1 month. In addition to skin lesions of lepromatous leprosy (subpolar type), there was nontender, non-fluctuant swelling of the right fifth and fourth fingers and left second finger. Skin over the right fifth finger showed sinus-like openings with associated purulent discharge. He also had swelling and ulceration of second left toe. Slit-skin smear (SSS) showed a bacterial index of 6+ from the ear lobes and cutaneous nodules, 4+ from the patch, and 3+ from normal skin. Modified Ziehl-Neelsen staining of the discharge extruding from the sinuses on the right fifth finger also showed abundant acid fast bacilli. Radiography of the hands and feet showed lytic lesions in the distal epimetaphyseal region o proximal phalanx of the right fifth finger and left second finger and erosion of distal end of proximal phalanges of both second toes. Histopathological examination of biopsy specimen from the patch (back) showed features of lepromatous leprosy, and Fite-Faraco stain for tissue acid fast bacteria (AFB) was strongly positive. Fine-needle-aspiration cytology (FNAC) from the lytic lesion in the bone also showed predominantly foamy macrophages with strongly positive staining for AFB with a few interspersed lymphocytes, epithelioid cells and Langhans giant cells. On the basis of these features, a clinical diagnosis of subpolar lepromatous leprosy with leprous osteitis was made. In today's clinical era of improved case detection and prompt treatment with effective multidrug regimens, advanced bone changes are rarely encountered. We describe this case of lepromatous leprosy that developed cavitating lesions of the phalanges of the hand, seen on x-ray as well-defined bone cyst and erosions. PMID- 15347500 TI - Coexisting basal cell carcinoma and metastatic small cell carcinoma of lung. AB - The coexistence of basal cell carcinoma with a cutaneous metastasis is rare; it is most frequently associated with lung cancer. We report an unusual case of contralateral metachronous metastasis of small-cell carcinoma of the lung to a site of basal cell carcinoma. PMID- 15347501 TI - Ant-induced alopecia: report of 2 cases and review of the literature. AB - Localized scalp hair loss is associated with many processes, including alopecia areata, trichotillomania, tinea capitis, and early lupus erythematosus. There are several reports of localized alopecia after tick- and flea-bites and bee stings, but there are only two reports of ant-induced alopecia in the literature. We present two cases of alopecia induced by ants of genus Pheidole (species pallidula) and review the literature for insect-induced alopecia. Ant-induced alopecia should be considered in the differential diagnosis of localized sudden onset alopecia, at least in some geographic areas of the world. PMID- 15347502 TI - Cutaneous variant of angiokeratoma corporis diffusum associated with angiokeratoma circumscriptum. PMID- 15347503 TI - Acrokeratosis paraneoplastica (Bazex syndrome): an atypical presentation. AB - A 62-year-old male presented with a 2-year history of hyperkeratotic lesions of the hands and feet. Previous treatment with topical steroids was unsuccessful. A complete physical examination revealed the presence of blood in the stool, and sigmoidoscopy showed an ulcerative growth at the rectosigmoid junction. The histopathology showed adenocarcinoma. PMID- 15347504 TI - Prescribing antidepressant drugs for pemphigus patients: An important point to keep in mind. PMID- 15347505 TI - Unusual localization of multiple myxoid (mucous) cysts of toes. AB - Myxoid cysts of fingers and toes are observed frequently on the lateral or dorsal aspects of the distal digits. They are usually solitary nodules. Both subungual localization and multiplicity are quite rare. We present a 74-year-old woman with digital subungual mucous cysts located on all toes. PMID- 15347506 TI - Treatment of lichen planopilaris with mycophenolate mofetil. AB - Mycophenolate mofetil (MMF) is an immunosuppressive drug that has recently been used to treat autoimmune and inflammatory skin diseases. We report the first case of lichen planopilaris (LPP) successfully treated with MMF. The treatment of our patient demonstrates a novel therapeutic option for patients with LPP; MMF treatment may be preferable to azathioprine treatment because MMF has a safer adverse-effect profile. Larger studies must be performed to establish the risk benefit ratio of various therapeutic dosages of MMF for these patients. PMID- 15347507 TI - The use of recombinant human epidermal growth factor (rhEGF) in a gentleman with drug-induced Steven Johnson syndrome. AB - A case of drug-induced Steven Johnson syndrome in a gentleman is reported. Its course of treatment with rhEGF was compared to conventional treatment in historical control. PMID- 15347508 TI - Age and growth dynamics of Tyrannosaurus rex. AB - Tyrannosaurus rex is the most commonly found North American latest Cretaceous theropod, but until the 1980s only five specimens had been discovered, and no more than six have received a full description. Consequently there has been little information on how old Tyrannosaurus specimens were at maturity or death. Histological analysis of seven individuals provided, for the first time, an opportunity to assess the age represented by the bone cortex, to estimate the average individual age of these skeletons, to determine whether they represented fully grown individuals, and to predict their individual longevity. Though a range of ages (15-25 years) was found for the specimens studied, the seven individuals demonstrate that T. rex reached effectively full size in less than 20 years. The growth rate of T. rex was comparable to that of the African elephant, which has a similar mass and time to maturity. Some of the known specimens of T. rex did not quite reach full size; others do not seem to have survived long after achieving it. PMID- 15347509 TI - Ice sheets promote speciation in boreal birds. AB - The premise that Pleistocene ice ages played an important role in generating present-day species diversity has been challenged by genetic data indicating that most of the youngest terrestrial species on Earth coalesced long before major glacial advances. However, study has been biased towards faunas distributed at low latitudes that were not directly fragmented by advancing ice sheets. Using mitochondrial sequence divergence and a molecular clock, we compared the coalescence times of pairs of avian species belonging to superspecies complexes from the high-latitude boreal forest with those of sub-boreal and tropical avifaunas of the New World. Remarkably, all coalescence events in boreal superspecies date to the Pleistocene, providing direct evidence that speciation was commonly initiated during recent glacial periods. A pattern of endemism in boreal superspecies plausibly links the timing of divergence to the fragmentation of the boreal forest by ice sheets during the Mid- and Late Pleistocene. In contrast to the boreal superspecies, only 56% of sub-boreal and 46% of tropical superspecies members coalesced during the Pleistocene, suggesting that avifaunas directly fragmented by ice sheets experienced rapid rates of diversification, whereas those distributed farther south were affected to a lesser extent. One explanation for the absence of pre-Pleistocene superspecies in boreal avifaunas is that strong selection pressures operated in boreal refugia, causing superspecies members to achieve ecological differentiation at an accelerated rate. PMID- 15347510 TI - Cross-Cordillera exchange mediated by the Panama Canal increased the species richness of local freshwater fish assemblages. AB - Completion of the Panama Canal in 1914 breached the continental divide and set into motion a natural experiment of unprecedented magnitude by bringing previously isolated freshwater fish communities into contact. The construction of a freshwater corridor connecting evolutionarily isolated communities in Pacific and Caribbean watersheds dramatically increased the rate of dispersal, without directly affecting species interactions. Here, we report that a large fraction of species have been able to establish themselves on the other side of the continental divide, whereas no species have become extinct, leading to a local increase in species richness. Our results suggest that communities are not saturated and that competitive exclusion does not occur over the time-scale previously envisioned. Moreover, the results of this unintentional experiment demonstrate that community composition and species richness were regulated by the regional process of dispersal, rather than by local processes such as competition and predation. PMID- 15347511 TI - Can sunspot activity and ultraviolet-B radiation explain cyclic outbreaks of forest moth pest species? AB - Cyclic outbreaks of forest moth pest species have long remained a puzzle for foresters and ecologists. This paper presents time-series exhibiting a strong negative relationship between sunspot numbers and population indices of autumnal and winter moths, both in a mountain birch forest in central Norway and in a mixed lowland forest in southern Norway. In the latter area, also the population level of a moth species feeding entirely on lichens was negatively related to sunspot numbers. Low sunspot activity leads to a thinner ozone layer and thus higher surface ultraviolet (UV)-B radiation. As winter moth larvae prefer leaves subjected to enhanced UV-B radiation, we suggest that the causal relationship between sunspots and moths is that the metabolic costs of producing UV-B protective pigments during periods of low sunspot activity reduce trees' and lichens' resistance to herbivores, and thus increase the survival of moth larvae. Higher peak densities of moth cycles in mountain forests could be explained by the general higher UV-B radiation at higher altitudes. PMID- 15347512 TI - Female blue tits adjust parental effort to manipulated male UV attractiveness. AB - The differential allocation hypothesis predicts that parents should adjust their current investment in relation to perceived mate attractiveness if this affects offspring fitness. It should be selectively advantageous to risk more of their future reproductive success by investing heavily in current offspring of high reproductive value but to decrease investment if offspring value is low. If the benefits of mate attractiveness are limited to a particular offspring sex we would instead expect relative investment in male versus female offspring to vary with mate attractiveness, referred to as 'differential sex allocation'. We present strong evidence for differential allocation of parental feeding effort in the wild and show an immediate effect on a component of offspring fitness. By experimentally reducing male UV crown coloration, a trait known to indicate attractiveness and viability in wild-breeding blue tits (Parus caeruleus), we show that females, but not males, reduce parental feeding rates and that this reduces the skeletal growth of offspring. However, differential sex allocation does not occur. We conclude that blue tit females use male UV coloration as an indicator of expected offspring fitness and adjust their investment accordingly. PMID- 15347513 TI - Spatially structured population dynamics in feral oilseed rape. AB - We studied the population dynamics of feral oilseed rape (Brassica napus) for 10 years (1993-2002) in 3658 adjacent permanent 100 m quadrats in the verges of the M25 motorway around London, UK. The aim was to determine the relative importance of different factors affecting the observed temporal patterns of population dynamics and their spatial correlations. A wide range of population dynamics was observed (downward or upward trends, cycles, local extinctions and recolonizations), but overall the populations were not self-replacing (lambda < 1). Many quadrats remained unoccupied throughout the study period, but a few were occupied at high densities for all 10 years. Most quadrats showed transient oilseed rape populations, lasting 1-4 years. There were strong spatial patterns in mean population density, associated with soil conditions and the successional age of the plant community dominating the verge, and these large-scale spatial patterns were highly consistent from year to year. The importance of seed spilled from trucks in transit to the processing plant at Erith in Kent was confirmed: rape populations were significantly higher on the 'to Erith' verge than the 'from Erith' verge (overall mean 2.83-fold greater stem density). Quadrats in which lambda > 1 were much more frequent in the 'to Erith' verge, indicating that seed immigration can give the spurious impression of self-replacing population dynamics in time-series analysis. There was little evidence of a pervasive Moran effect, and climatic forcing did not produce widespread large-scale synchrony in population dynamics for the motorway as a whole; just 23% of quadrats had significant rank correlations with the mean time-series. There was, however, significant local spatial synchrony of population dynamics, apparently associated with soil disturbance and seed input. This study draws attention to the possibility that different processes may impose population synchrony at different scales. We hypothesize that synchrony in this system is driven by at least three processes: small-scale, local forcing caused by soil disturbance, intermediate scale forcing as a result of seed input, and large-scale climatic forcing (e.g. winter rainfall) that affects the motorway as a whole. PMID- 15347514 TI - Evolution of spite through indirect reciprocity. AB - How can cooperation persist in the face of a temptation to 'cheat'? Several recent papers have suggested that the answer may lie in indirect reciprocity. Altruistic individuals may benefit by eliciting altruism from observers, rather than (as in direct reciprocity) from the recipient of the aid they provide. Here, we point out that indirect reciprocity need not always favour cooperation; by contrast, it may support spiteful behaviour, which is costly for the both actor and recipient. Existing theory suggests spite is unlikely to persist, but we demonstrate that it may do so when spiteful individuals are less likely to incur aggression from observers (a negative form of indirect reciprocity). PMID- 15347515 TI - Prenatal developmental conditions have long-term effects on offspring fecundity. AB - Maternal effects, in which differences in parental state cause differences in offspring fitness, are important in trade-offs influencing an individual's optimal reproductive strategy. In zebra finches (Taeniopygia guttata) we manipulated the nutritional state for four weeks before the start of breeding through protein supplementation. Zebra finches were kept on identical diets during the rest of the experiment. We then tested the effects of maternal state on offspring size, survival and fecundity. In order to separate the effects of maternal state occurring through egg production, incubation and chick-rearing, we used a cross-fostering experiment. We show that a protein-rich diet prior to laying improved maternal body weight prior to breeding compared with birds on a protein-poor diet. Poorer maternal state prior to breeding gave rise to offspring with lower fecundity than offspring from birds in a better nutritional state. Maternal state is thought to affect the conditions developing offspring experience through the bird's ability to produce and incubate eggs. Male and female embryos differed in their responses to conditions at different developmental stages. This shows that embryonic developmental conditions and sex differences in vulnerability to these conditions need to be incorporated into future models of selection, life-history evolution and sex-ratio theory. PMID- 15347516 TI - Female starlings adjust primary sex ratio in response to aromatic plants in the nest. AB - Adjustment of offspring sex ratios should be favoured by natural selection when parents are capable of facultatively altering brood sex ratios and of recognizing the circumstances that predict the probable fitness benefit of producing sons and daughters. Although experimental studies have shown that female birds may adjust offspring sex ratios in response to changes in their own condition and in the external appearance of their mate, and male attributes other than his external morphology are also thought to act as signals of male quality, it is not known whether females will respond to changes in such signals, in the absence of any change in the appearance of the male himself. Here, we experimentally manipulated a male courtship display, the green plants carried to the nest by male spotless starlings (Sturnus unicolor), without changing any physical attributes of the male himself, and examined whether this influenced female decisions on offspring sex ratio. We found that in an environment in which female starlings were producing more daughters than sons, experimental enhancement of the green nesting material caused females to significantly increase the number of male eggs produced and thereby removed the female bias. This effect was consistent in 2 years and at two localities. This demonstrates that the green material, whose function has long puzzled biologists, conveys important information to the female and that she facultatively adjusts offspring production accordingly. PMID- 15347517 TI - Soil moisture and sex ratio in a plant with nuclear-cytoplasmic sex inheritance. AB - I investigated whether soil moisture affects relative fitness of females and hermaphrodites and sex ratio in a gynodioecious plant with nuclear-cytoplasmic sex inheritance. I contrast these results with those from species with strictly nuclear sex inheritance. I performed a manipulative watering experiment on seed fitness of the two sexes, and field studies measuring seed fitness and sex ratio as a function of soil moisture. In the dry site, watered hermaphrodites produced approximately twice as many seeds as unwatered hermaphrodites, with little treatment effect on female seed production. Over a natural soil moisture gradient, the ratio of female to hermaphrodite seed production was higher in dry than in wet sites. These data show that the seed fitness advantage of females is a function of soil moisture. Despite this, regression of soil moisture on the sex ratio of 23 populations was not significant. These results indicate a sex dependent effect of soil moisture on resource allocation to seeds that does not translate into a strong effect on sex ratio. This is consistent with theory based on genomic conflict in which sex ratios are predicted to be only partly determined by fitness differences of the sexes. PMID- 15347518 TI - A cost of Wolbachia-induced sex reversal and female-biased sex ratios: decrease in female fertility after sperm depletion in a terrestrial isopod. AB - A number of parasites are vertically transmitted to new host generations via female eggs. In such cases, host reproduction is an intimate component of parasite fitness and no cost of the infection on host reproduction is expected to evolve. A number of these parasites distort host sex ratios towards females, thereby increasing either parasite fitness or the proportion of the host that transmit the parasite. In terrestrial isopods (woodlice), Wolbachia bacteria are responsible for sex reversion and female-biased sex ratios, changing genetic males into functional neo-females. Although sex ratio distortion is a powerful means for parasites to increase in frequency in host populations, it also has potential consequences on host biology, which may, in turn, have consequences for parasite prevalence. We used the woodlouse Armadillidium vulgare to test whether the interaction between Wolbachia infection and the resulting excess of females would limit female fertility through the reduction in sperm number that they receive from males. We showed that multiple male mating induces sperm depletion, and that this sperm depletion affects fertility only in infected females. This decrease in fertility, associated with male mate choice, may limit the spread of Wolbachia infections in host populations. PMID- 15347519 TI - Evolutionary trade-off between defence against grazing and competitive ability in a simple unicellular alga, Chlorella vulgaris. AB - Trade-offs between defence and other fitness components are expected in principle, and can have major qualitative impacts on ecological dynamics. Here we show that such a trade-off exists even in the simple unicellular alga Chlorella vulgaris. We grew algal populations for multiple generations in either the presence ('grazed algae') or absence ('non-grazed algae') of the grazing rotifer Brachionus calyciflorus, and then evaluated their defence and competitive abilities. Grazed algae were better defended, yielding rotifer growth rate 32% below that of animals fed non-grazed algae, but they also had diminished competitive ability, with a growth rate under nutrient-limiting conditions 28% below that of non-grazed algae. Grazed algae also had a smaller cell size and were more concentrated in carbon and nitrogen. Thus, C. vulgaris genotypes vary phenotypically in their position along a trade-off curve between defence against grazing and competitive ability. This genetic variation underlies rapid algal evolution that significantly alters the ecological predator-prey cycles between rotifers and algae. PMID- 15347520 TI - Complex social behaviour can select for variability in visual features: a case study in Polistes wasps. AB - The ability to recognize individuals is common in animals; however, we know little about why the phenotypic variability necessary for individual recognition has evolved in some animals but not others. One possibility is that natural selection favours variability in some social contexts but not in others. Polistes fuscatus wasps have variable facial and abdominal markings used for individual recognition within their complex societies. Here, I explore whether social behaviour can select for variability by examining the relationship between social behaviour and variability in visual features (marking variability) across social wasp taxa. Analysis using a concentrated changes test demonstrates that marking variability is significantly associated with nesting strategy. Species with flexible nest-founding strategies have highly variable markings, whereas species without flexible nest-founding strategies have low marking variability. These results suggest that: (i) individual recognition may be widespread in the social wasps, and (ii) natural selection may play a role in the origin and maintenance of the variable distinctive markings. Theoretical and empirical evidence suggests that species with flexible nesting strategies have reproductive transactions, a type of complex social behaviour predicted to require individual recognition. Therefore, the reproductive transactions of flexible species may select for highly variable individuals who are easy to identify as individuals. Further, selection for distinctiveness may provide an alternative explanation for the evolution of phenotypic diversity. PMID- 15347521 TI - Cumulative meta-analysis: a new tool for detection of temporal trends and publication bias in ecology. AB - Temporal changes in the magnitude of research findings have recently been recognized as a general phenomenon in ecology, and have been attributed to the delayed publication of non-significant results and disconfirming evidence. Here we introduce a method of cumulative meta-analysis which allows detection of both temporal trends and publication bias in the ecological literature. To illustrate the application of the method, we used two datasets from recently conducted meta analyses of studies testing two plant defence theories. Our results revealed three phases in the evolution of the treatment effects. Early studies strongly supported the hypothesis tested, but the magnitude of the effect decreased considerably in later studies. In the latest studies, a trend towards an increase in effect size was observed. In one of the datasets, a cumulative meta-analysis revealed publication bias against studies reporting disconfirming evidence; such studies were published in journals with a lower impact factor compared to studies with results supporting the hypothesis tested. Correlation analysis revealed neither temporal trends nor evidence of publication bias in the datasets analysed. We thus suggest that cumulative meta-analysis should be used as a visual aid to detect temporal trends and publication bias in research findings in ecology in addition to the correlative approach. PMID- 15347522 TI - A phylogenetic review of the African leaf chameleons: genus Rhampholeon (Chamaeleonidae): the role of vicariance and climate change in speciation. AB - The phylogenetic associations among 13 currently recognized African leaf chameleon species were investigated by making use of mitochondrial and nuclear DNA sequence data (44 taxa and 4145 characters). The gene tree indicates two divergent clades within Rhampholeon; this finding is congruent with previous morphological suggestions. The first clade (I) comprises three taxa (R. kerstenii, R. brevicaudatus and R. brachyurus) and is widely distributed in lowland forest and or non-forest biomes. The second clade (II) comprises the remaining Rhampholeon species and can be subdivided into three subclades. By contrast, most taxa belonging to clade II are confined to relict montane forest biotopes. Based on geographical, morphological and molecular evidence, it is suggested that the taxonomy of Rhampholeon be revised to include two genera (Rieppeleon and Rhampholeon) and three subgenera (Rhampholeon, Bicuspis and Rhinodigitum). There is a close correlation between geographical distribution and phylogenetic relatedness among Rhampholeon taxa, indicating that vicariance and climate change were possibly the most influential factors driving speciation in the group. A relaxed Bayesian clock suggests that speciation times coincided both with the northern movement of Africa, which caused the constriction of the pan African forest, and to rifting in east Africa ca. 20 Myr ago. Subsequent speciation among taxa was probably the result of gradual desiccation of forests between 20 and 5 Myr ago. PMID- 15347523 TI - Biodiversity and the Lotka-Volterra theory of species interactions: open systems and the distribution of logarithmic densities. AB - Theoretical interest in the distributions of species abundances observed in ecological communities has focused recently on the results of models that assume all species are identical in their interactions with one another, and rely upon immigration and speciation to promote coexistence. Here we examine a one-trophic level system with generalized species interactions, including species-specific intraspecific and interspecific interaction strengths, and density-independent immigration from a regional species pool. Comparisons between results from numerical integrations and an approximate analytic calculation for random communities demonstrate good agreement, and both approaches yield abundance distributions of nearly arbitrary shape, including bimodality for intermediate immigration rates. PMID- 15347524 TI - Introduction: why do we need a science of well-being? AB - INTRODUCTION: why do we need a science of well-being? PMID- 15347525 TI - Natural selection and the elusiveness of happiness. AB - The quest for happiness has expanded from a focus on relieving suffering to also considering how to promote happiness. However, both approaches have yet to be conducted in an evolutionary framework based on the situations that shaped the capacities for happiness and sadness. Because of this, the emphasis has almost all been on the disadvantages of negative states and the benefits of positive states, to the nearly total neglect of 'diagonal psychology', which also considers the dangers of unwarranted positive states and the benefits of negative emotions in certain situations. The situations that arise in goal pursuit contain adaptive challenges that have shaped domain-general positive and negative emotions that were partially differentiated by natural selection to cope with the more specific situations that arise in the pursuit of different kinds of goals. In cultures where large social groups give rise to specialized and competitive social roles, depression may be common because regulation systems are pushed far beyond the bounds for which they were designed. Research on the evolutionary origins of the capacities for positive and negative emotions is urgently needed to provide a foundation for sensible decisions about the use of new mood manipulating technologies. PMID- 15347526 TI - Understanding well-being in the evolutionary context of brain development. AB - Much of the work on well-being and positive emotions has tended to focus on the adult, partly because this is when problems are manifest and well-being often becomes an issue by its absence. However, it is pertinent to ask if early life events might engender certain predispositions that have consequences for adult well-being. The human brain undergoes much of its growth and development postnatally until the age of seven and continues to extend its synaptic connections well into the second decade. Indeed, the prefrontal association cortex, areas of the brain concerned with forward planning and regulatory control of emotional behaviour, continue to develop until the age of 20. In this article, I consider the significance of this extended postnatal developmental period for brain maturation and how brain evolution has encompassed certain biological changes and predispositions that, with our modern lifestyle, represent risk factors for well-being. An awareness of these sensitive phases in brain development is important in understanding how we might facilitate secure relationships and high self-esteem in our children. This will provide the firm foundations on which to develop meaningful lifestyles and relationships that are crucial to well-being. PMID- 15347527 TI - The developmental origins of well-being. AB - Low birthweight is now known to be associated with increased rates of coronary heart disease and the related disorders, stroke, hypertension and adult-onset diabetes. These associations have been extensively replicated in studies in different countries and are not the result of confounding variables. They extend across the normal range of birthweight and depend on lower birthweights in relation to the duration of gestation rather than the effects of premature birth. The associations are thought to be consequences of developmental plasticity, the phenomenon by which one genotype can give rise to a range of different physiological or morphological states in response to different environmental conditions during development. Recent observations have shown that impaired growth in infancy and rapid childhood weight gain exacerbate the effects of impaired prenatal growth. A new vision of optimal early human development is emerging, which takes account of health and well-being throughout life. PMID- 15347528 TI - The broaden-and-build theory of positive emotions. AB - The broaden-and-build theory describes the form and function of a subset of positive emotions, including joy, interest, contentment and love. A key proposition is that these positive emotions broaden an individual's momentary thought-action repertoire: joy sparks the urge to play, interest sparks the urge to explore, contentment sparks the urge to savour and integrate, and love sparks a recurring cycle of each of these urges within safe, close relationships. The broadened mindsets arising from these positive emotions are contrasted to the narrowed mindsets sparked by many negative emotions (i.e. specific action tendencies, such as attack or flee). A second key proposition concerns the consequences of these broadened mindsets: by broadening an individual's momentary thought-action repertoire--whether through play, exploration or similar activities--positive emotions promote discovery of novel and creative actions, ideas and social bonds, which in turn build that individual's personal resources; ranging from physical and intellectual resources, to social and psychological resources. Importantly, these resources function as reserves that can be drawn on later to improve the odds of successful coping and survival. This chapter reviews the latest empirical evidence supporting the broaden-and-build theory and draws out implications the theory holds for optimizing health and well-being. PMID- 15347529 TI - A balanced psychology and a full life. AB - Psychology since World War II has been largely devoted to repairing weakness and understanding suffering. Towards that end, we have made considerable gains. We have a classification of mental illness that allows international collaboration, and through this collaboration we have developed effective psychotherapeutic or pharmacological treatments for 14 major mental disorders. However, while building a strong science and practice of treating mental illness, we largely forgot about everyday well-being. Is the absence of mental illness and suffering sufficient to let individuals and communities flourish? Were all disabling conditions to disappear, what would make life worth living? Those committed to a science of positive psychology can draw on the effective research methods developed to understand and treat mental illness. Results from a new randomized, placebo controlled study demonstrate that people are happier and less depressed three months after completing exercises targeting positive emotion. The ultimate goal of positive psychology is to make people happier by understanding and building positive emotion, gratification and meaning. Towards this end, we must supplement what we know about treating illness and repairing damage with knowledge about nurturing well-being in individuals and communities. PMID- 15347530 TI - Positive health: connecting well-being with biology. AB - Two key types of well-being, eudaimonic and hedonic, are reviewed. The first addresses ideas of self-development, personal growth and purposeful engagement, while the second is concerned with positive feelings such as happiness and contentment. How well-being varies by age and socio-economic standing is briefly summarized, followed by examination of its biological correlates (neuroendocrine, immune, cardiovascular, rapid eye movement (REM) sleep). Preliminary findings on a sample of ageing women showed that those with higher levels of eudaimonic well being had lower levels of daily salivary cortisol, pro-inflammatory cytokines, cardiovascular risk, and longer duration REM sleep compared with those showing lower levels of eudaimonic well-being. Hedonic well-being, however, showed minimal linkage to biomarker assessments. Future research directions building on these initial findings are discussed. PMID- 15347531 TI - Well-being and affective style: neural substrates and biobehavioural correlates. AB - One of the most salient features of emotion is the pronounced variability among individuals in their reactions to emotional incentives and in their dispositional mood. Collectively, these individual differences have been described as affective style. Recent research has begun to dissect the constituents of affective style. The search for these components is guided by the neural systems that instantiate emotion and emotion regulation. In this article, this body of research and theory is applied specifically to positive affect and well-being. The central substrates and peripheral biological correlates of well-being are described. A resilient affective style is associated with high levels of left prefrontal activation, effective modulation of activation in the amygdala and fast recovery in response to negative and stressful events. In peripheral biology, these central patterns are associated with lower levels of basal cortisol and with higher levels of antibody titres to influenza vaccine. The article concludes with a consideration of whether these patterns of central and peripheral biology can be modified by training and shifted toward a more salubrious direction. PMID- 15347532 TI - Successful ageing: from cell to self. AB - Many people see ageing as a time of cognitive and physical decline. For the past three decades, most scientists and the general public have accepted this negative age-stereotype as the norm, but fortunately this view is now challenged. New findings show that well-being and a positive view of ageing are major protective factors against the effects of age on the organism. These results challenge the scientific studies that place emphasis on the negative side of ageing. This ageism view has been observed in each sphere of science, from genetics to social sciences. Perspectives from each domain are described, and new integrative views of successful ageing are summarized. PMID- 15347533 TI - Intelligence and culture: how culture shapes what intelligence means, and the implications for a science of well-being. AB - This paper discusses the relationship between culture and intelligence. The main message of the paper is that intelligence cannot fully or even meaningfully be understood outside its cultural context. Behaviour that is considered intelligent in one culture may be considered unintelligent in another culture, and vice versa. Moreover, people in different cultures have different implicit (folk) theories of intelligence, so may not even mean the same thing by the word. The relationships between different aspects of intelligence can vary across cultures, with correlations that are positive in one setting proving to be negative in another. The paper opens with a general discussion of issues regarding the relationship between the two concepts. It then describes the theory of successful intelligence, which motivates our work on the interface between culture and intelligence. Finally, the article draws some conclusions. PMID- 15347534 TI - The social context of well-being. AB - Large samples of data from the World Values Survey, the US Benchmark Survey and a comparable Canadian survey are used to estimate equations designed to explore the social context of subjective evaluations of well-being, of happiness, and of health. Social capital, as measured by the strength of family, neighbourhood, religious and community ties, is found to support both physical health and subjective well-being. Our new evidence confirms that social capital is strongly linked to subjective well-being through many independent channels and in several different forms. Marriage and family, ties to friends and neighbours, workplace ties, civic engagement (both individually and collectively), trustworthiness and trust: all appear independently and robustly related to happiness and life satisfaction, both directly and through their impact on health. PMID- 15347535 TI - Well-being: towards an integration of psychology, neurobiology and social science. AB - Well-being: towards an integration of psychology, neurobiology and social science PMID- 15347536 TI - Length of stay and interval to readmission in emergency hospital treatment of COPD. AB - BACKGROUND: increasing attention has recently been attached to the length of hospital stay and related factors in the treatment of COPD. OBJECTIVES: to assess the trend in the duration of inpatient episodes following emergency admissions for COPD by age and sex, and the frequency of readmissions, as well as the correlations between the frequency and duration of inpatient episodes. DESIGN: retrospective study. SETTING: the Finnish hospital discharge register. SUBJECTS: the 72,672 inpatient episodes following emergency admissions of patients aged over 44 years that ended in 1993-2001 and had COPD as the principal diagnosis. RESULTS: the mean duration of inpatient episodes was 8.5 days (SD 8.2) in 1993, but 6.8 days (SD 6.6) in 2001. The figure for 45- to 64-year-old men was 6.5 days (SD 6.6) and that for men aged >64 years, 7.8 days (SD 6.8). The corresponding figures for women were 7.1 days (SD 6.8) and 8.8 days (SD 8.4). The average interval between the end of one inpatient episode and the beginning of the next was 195.4 days (SD 327.7). This interval was longest when the inpatient episode lasted for 7 days (interval 215 days). CONCLUSIONS: the length of hospital stay for COPD exacerbation seems to be decreasing, and elderly women have the longest inpatient episodes. With the current treatment modalities, a 1-week stay in hospital results in the longest interval to readmission. The situation may change if supported home care at exacerbation can be increased. PMID- 15347537 TI - Can adapting the homes of older people and providing assistive technology pay its way? AB - BACKGROUND: Adaptations and assistive technology (AT) have an important role in enabling older people to remain in their own homes. OBJECTIVE: To measure the feasibility and cost of adaptations and AT, and the scope for these to substitute and supplement formal care. DESIGN: Detailed design studies to benchmark the adaptability of 82 properties against the needs of seven notional users. SETTING: Social rented housing sector. MAIN OUTCOME MEASURES: Measures of the adaptability of properties, costs of care, adaptations and AT, and relationships between these costs. RESULTS: The adaptability of properties varies according to many design factors and the needs of occupiers. The most adaptable properties were ground floor flats and bungalows; the least were houses, maisonettes and flats in converted houses. Purpose-built sheltered properties were generally more adaptable than corresponding mainstream properties but the opposite was the case for bungalows. Adaptations and AT can substitute for and supplement formal care, and in most cases the initial investment in adaptations and AT is recouped through subsequently lower care costs within the average life expectancy of a user. CONCLUSION: Appropriately selected adaptations and AT can make a significant contribution to the provision of living environments which facilitate independence. They can both substitute for traditional formal care services and supplement these services in a cost-effective way. PMID- 15347538 TI - Emergency medical services utilization during an outbreak of severe acute respiratory syndrome (SARS) and the incidence of SARS-associated coronavirus infection among emergency medical technicians. AB - OBJECTIVES: This was a study to evaluate the utilization of emergency medical services (EMS) systems during the outbreak of severe acute respiratory syndrome (SARS), and to assess the incidence of infection among emergency medical technicians (EMTs). METHODS: This was a prospective, observational study conducted in the EMS system of Taipei, Taiwan. Probable/suspect cases of SARS were defined by World Health Organization criteria. SARS-related transports were categorized into 1) requests from hospitals for probable/suspect cases of SARS, 2) quarantined individuals, and 3) febrile persons. City ambulances were organized into teams A, B, and C for transports of different perceived risks. Data on the EMS volume, the transport category, the final SARS status of patients, and the EMT responsible for the transports were collected. The EMS projected volume was computed by previous years' data and compared with that collected. The SARS incidence among EMTs was assessed by investigating probable SARS (P-SARS) and by surveying the seroprevalence of SARS-associated coronavirus (SARS-CoV) antibody. RESULTS: From March 18 to June 19, 2003, there were 7,961 EMS transports, similar to the volume projected from previous years (7,506) (95% CI = 6,688 to 8,324). Of these, 1,760 (22.1%) were SARS-related. When SARS related transports were excluded, there was a 12.2% decrease (95% CI = 11.4% to 12.9%) in EMS activities. Requests from hospitals, quarantined individuals, and febrile citizens accounted for 23%, 18%, and 59% of SARS-related transports. Among the 397 P-SARS cases in the city of 2.65 million people (incidence 0.01%, 95% CI = 0.01% to 0.02%), 138 (35%) required EMS transports. Two EMTs working in team C, the team with the lowest risk, developed P-SARS. One of them died soon thereafter. The incidence of P-SARS was 0.6% (95% CI = 0.2% to 2.2%), or 0.1% (95% CI = 0.03% to 0.4%) per transport. SARS-CoV serology was available in 74.1% of EMTs who were alive. In addition to the surviving P-SARS EMT, one EMT from team A, the team with the highest risk, was seropositive. Combining P-SARS and the seropositive case, three EMTs were infected (incidence 1.3%, 95% CI = 0.4% to 3.6%). No patient transported by the infected EMTs developed SARS. The hospitals serving EMS by the infected EMTs had been involved in a clustered outbreak prior to the EMTs' infections. CONCLUSIONS: During the outbreak of SARS, the overall EMS volume did not change significantly, but the non-SARS EMS activities decreased. Compared with the general population, EMS providers are at higher risk of contracting the SARS virus regardless of different perceived levels of risk. Standard protections and procedures for infection control should be strictly followed during transport and within the hospital environment. PMID- 15347539 TI - Outcome of patients with an indeterminate emergency department first-trimester pelvic ultrasound to rule out ectopic pregnancy. AB - OBJECTIVES: Pelvic ultrasound performed by emergency physicians can identify a definite diagnosis in the majority of symptomatic first-trimester pregnant patients on the initial emergency department (ED) visit. However, a significant minority of such patients are diagnosed as having an indeterminate pregnancy state requiring further testing and consultation. The authors investigated the final outcome of patients with an initial indeterminate ED first-trimester pelvic ultrasound examination in the setting of an interdepartmental protocol to rule out ectopic pregnancy. METHODS: This was an observational prospective cohort study performed at a regional, urban ED with more than 100,000 patient visits over a 13-month period. Pelvic ultrasound for first-trimester patients was prospectively performed by emergency physicians with gynecologic consultation for lack of intrauterine pregnancy (IUP) ultrasound findings. IUP was defined as a fundal gestational sac with either a yolk sac or a fetal pole. Pelvic ultrasounds were classified into diagnostic categories including definite IUP, embryonic demise, molar pregnancy, definite ectopic pregnancy, and indeterminate. For all patients with indeterminate pelvic ultrasound findings, final diagnostic categories and patient outcome were established by the use of patient records, obstetric ultrasound reports, laboratory studies, operative reports, and pathology reports. All patients with ectopic pregnancy were followed for mode of treatment. Descriptive statistics were calculated. RESULTS: A total of 1,490 ED first-trimester pelvic ultrasound examinations were performed over 13 months establishing the following diagnostic rates for initial ED visit: IUP 1,037 (70%), demise 127 (8%), definite ectopic pregnancy 24 (2%), molar pregnancy 2 ( < 1%), and indeterminate 300 (20%). The 300 indeterminate patients were classified using the above protocol into the following final diagnostic categories: embryonic demise 158 (53%), IUP 88 (29%), ectopic pregnancy 44 (15%), and unknown outcome 10 (3%). Indeterminate patients with ectopic pregnancy were treated with methotrexate in 25 of 44 cases (57%) and surgically in 16 of 44 cases (36%); there were no laparotomies. In contrast, ectopic pregnancy patients diagnosed on initial ED visit were treated surgically in 20 of 24 cases (83%), including four laparotomies. CONCLUSIONS: The outcome of symptomatic first-trimester patients with indeterminate ED pelvic ultrasounds is poor, with significantly high rates of embryonic demise and ectopic pregnancy. However, those indeterminate patients with the eventual diagnosis of ectopic pregnancy have a higher rate of medical methotrexate treatment and a reduced rate of invasive surgical treatment compared with ectopic pregnancy patients diagnosed at initial ED visit. PMID- 15347540 TI - A randomized clinical trial to assess the efficacy of the Epley maneuver in the treatment of acute benign positional vertigo. AB - OBJECTIVES: To compare the efficacy of the Epley maneuver with that of a placebo maneuver in patients presenting to the emergency department (ED) with benign positional vertigo (BPV). METHODS: This was a prospective, randomized, single blind placebo-controlled trial. Consecutive adult ED patients presenting to a university teaching hospital with BPV were randomized to treatment with either the Epley or placebo maneuver. The severity of vertigo was evaluated on a 0 to 10 point scale before and after the maneuvers. RESULTS: Eleven patients were randomized to the Epley group and 11 to the placebo group before the trial was terminated, based on a planned interim analysis. The median decreases in vertigo severity were 6 (95% confidence interval [95% CI] = 4 to 9) for the Epley group and 1 (95% CI = 0 to 3) for the placebo group (p = 0.001). CONCLUSIONS: The Epley maneuver is a simple bedside maneuver that appears to be more efficacious than a placebo maneuver in the treatment of acute BPV among ED patients. PMID- 15347541 TI - Glasgow Coma Scale score and QTc interval in the prognosis of organophosphate poisoning. AB - OBJECTIVES: The aim of this study was to assess the applicability of the Glasgow Coma Scale (GCS) score and the Q-T interval corrected for heart rate (QTc interval) in predicting outcome and complications in patients with organophosphate (OP) poisoning. METHODS: This prospective, observational study included 65 patients older than 18 years. In the out-of-hospital setting, the end tidal carbon dioxide (ETCO2), oxygen saturation (SaO2), QTc interval, and GCS score were monitored in each patient. A statistical comparison was then made between the group with respiratory failure and the group without this complication. RESULTS: The group with complications had significantly different values of measured parameters--a longer QTc interval and a lower GCS score, a higher number of intubations, and worse outcomes (p < 0.05). The two measures, GCS score and QTc interval, have been shown to be equally good in predicting respiratory failure and hospital mortality in patients with OP poisoning. CONCLUSIONS: In the initial out-of-hospital care of patients with OP poisoning, it is essential to monitor QTc interval and GCS score. These measures help with prognosis, and may suggest when to initiate precautions to prevent complications (i.e., respiratory failure). The simplicity and promptness of these methods allow providers to perform early and effective triage. PMID- 15347542 TI - Assessment of resident professionalism using high-fidelity simulation of ethical dilemmas. AB - OBJECTIVES: To examine the responses of emergency medicine residents (EMRs) to ethical dilemmas in high-fidelity patient simulations as a means of assessing resident professionalism. METHODS: This cross-sectional observational study included all EMRs at a three-year training program. Subjects were excluded if they were unable or unwilling to participate. Each resident subject participated in a simulated critical patient encounter during an Emergency Medicine Crisis Resource Management course. An ethical dilemma was introduced before the end of each simulated encounter. Resident responses to that dilemma were compared with a professional performance checklist evaluation. Multi-response permutation procedure analysis was used to compare performance measures between resident classes, with the a priori hypothesis that mean performance should increase as experience increases. RESULTS: Of the 30 potential subjects, 90% (27) participated. The remaining three residents were unavailable due to scheduling conflicts. It was observed that senior residents (second and third year) performed more checklist items than did first-year residents (p < 0.028 for each senior class). Omnibus comparison of mean critical actions completed across all three years was not statistically significant (p < 0.13). Residents performed a critical action with 100% uniformity across training years in only one ethical scenario ("Practicing Procedures on the Recently Dead"). Residents performed the fewest critical actions and overall checklist items for the "Patient Confidentiality" case. CONCLUSIONS: Senior residents had better overall performance than incoming interns, suggesting that professional behaviors are learned through some facet of residency training. Although limited by small sample size, the application of this performance-assessment tool showed the ability to discriminate between experienced and inexperienced EMRs with respect to a variety of aspects of professional competency. These findings suggest a need for improved resident education in areas of professionalism and ethics. PMID- 15347543 TI - The effect of emergency medicine residency format on pursuit of fellowship training and an academic career. AB - OBJECTIVES: To determine the association between emergency medicine (EM) program format (postgraduate year [PGY] 1-3, 2-4, or 1-4) and two dependent variables: fellowship training and academic career. METHODS: The authors conducted a mailed survey of 122 program directors (PDs) of U.S. EM residencies regarding the number of graduates from 1995 to 2000 who pursued fellowships, community practice, academics, or an advanced degree. The survey asked for initial postresidency position, as well as position three to five years later. RESULTS: Of those contacted, 67.2% of the programs responded regarding 3,521 graduates (70.4% of all graduates); 18.6% of 1-3-year program graduates chose academics versus 28.5% for 2-4 and 34.2% for 1-4. A greater proportion of four-year-format graduates pursued academics (PGY 2-4 vs. PGY 1-3, odds ratio [OR] = 1.74; 95% confidence interval [95% CI] = 1.44 to 2.11, PGY 1-4 vs. PGY 1-3, OR = 2.28; 95% CI = 1.81 to 2.85). The PGY 1-4 format was associated with academic practice versus the PGY 2-4 format (OR = 1.31; 95% CI = 1.02 to 1.67). The aggregate of the PGY 2-4 and 1 4 formats was associated with initial academics versus PGY 1-3 programs (OR = 1.92; 95% CI = 1.63 to 2.26). Of PGY 1-3 residents, 4.3% pursued fellowships versus 5.6% of 2-4 and 8.6% of 1-4. The PGY 1-4 format was associated with more common fellowship pursuit versus both 2-4 (OR = 1.59; 95% CI = 1.01 to 2.51) and 1-3 (OR = 2.08; 95% CI = 1.41 to 3.10). For 1995-1997 graduates, 74.2% (271/365) who started out in academics remained there three to five years later. Of all graduates, 5.2% pursued fellowships and 23.1% pursued academics initially. Sixty seven of 271 (24.7%) academic physicians from the 1995-1997 classes were fellowship-trained. CONCLUSIONS: Four-year formats, especially 1-4, were associated with more common pursuit of fellowships and academics than the 1-3 format. Fellowship pursuit was uncommon (4% to 9% of graduates), whereas 18% to 34% initially chose academics. PMID- 15347544 TI - Predicting asthma morbidity in Harlem emergency department patients. AB - OBJECTIVES: To determine predictors of asthma morbidity in African American patients with asthma. Proxies for asthma morbidity were emergency department (ED) visits for asthma and hospitalizations for asthma. METHODS: This was a prospective observational study that evaluated baseline predictors of asthma morbidity in adults in an urban, predominantly African American community in New York City. Potential predictors of asthma morbidity evaluated were education, gender, employment status, current smoking status, asthma severity, duration of asthma, daily use of a peak flow meter, presence or absence of pets at home, presence or absence of a significant other, presence or absence of medical insurance, and previous hospitalization for asthma in the past year. Follow-up consisted of a repeat questionnaire obtained between nine and 15 months after the baseline questionnaire. Follow-up data collection was limited to the last three month history of ED visits or hospitalizations before the follow-up visit. At follow-up, the baseline predictors were related to the presence or absence of ED visits for asthma or hospitalizations for asthma. All predictors were evaluated individually (crude odds ratio [OR]) and simultaneously (adjusted OR) in a logistic regression model with the dichotomous outcome variable ED visits or hospitalization. RESULTS: Return ED visits on follow-up were more likely to occur in asthma patients hospitalized in the previous year (adjusted OR, 3.9; 95% confidence interval [CI] = 1.7 to 9.0) and were less likely to occur in asthma patients with pets (OR, 0.4; 95% CI = 0.2 to 0.9). Patients with moderate/severe asthma, relative to patients with mild asthma, were more likely to be seen in the ED on follow-up on initial analysis (crude OR, 2.4; 95% CI = 1.1 to 1.5), but the adjusted OR was not significant. Follow-up hospitalizations were significantly more likely to occur only in subjects reporting daily use of a peak flow meter (OR, 6.8; 95% CI = 1.3 to 34.5). Subjects hospitalized for asthma in the previous year were more likely to be hospitalized subsequently on initial analysis (crude OR, 2.9; 95% CI = 1.0 to 8.1), but the adjusted OR was not significant. CONCLUSIONS: It appears that African American patients with asthma who had previous hospitalizations for asthma within the past year or use a peak flow meter daily (a marker for more severe asthma) are more likely to visit the ED in the future or to be hospitalized for asthma, respectively. These patients need to be targeted and treated more aggressively to improve asthma care and decrease morbidity. The apparent protective effect of the presence of pets on reducing ED visits is unclear at this time, and the findings need to be replicated and evaluated further. PMID- 15347545 TI - Insurer's duty to settle: friend or foe? PMID- 15347546 TI - Advanced statistics: the propensity score--a method for estimating treatment effect in observational research. AB - Observational studies assessing the effect of a particular treatment or exposure may be subject to bias, which can be difficult to eliminate using standard analytic techniques. Multivariable models are commonly used in observational research to assess the relationship between a certain exposure or treatment and an outcome, while adjusting for important variables necessary to ensure comparability between the groups. Large differences in the observed covariates between two study groups may exist in observational studies in which the investigator has no control over who was allocated to each treatment group, and these differences may lead to biased estimates of treatment effect. When there are large differences in important prognostic characteristics between the treatment groups, adjusting for these differences with conventional multivariable techniques may not adequately balance the groups, and the remaining bias may limit valid causal inference. Use of a propensity score, described as a conditional probability that a subject will be "treated" based on an observed group of covariates, may better adjust covariates between the groups and reduce bias. The purpose of this article is to describe the use of propensity scores to adjust for bias when estimating treatment effects in observational research and to compare use of this technique with conventional multivariable regression. The authors present three methods for integrating propensity scores into observational analyses using a database collected on head-injured trauma patients. The article details the methods for creating a propensity score, analyzing data with the score, and explores differences between propensity score methods and conventional multivariable methods, including potential benefits and limitations. Graphical representations of the analyses are provided as well. PMID- 15347547 TI - Ethics seminars: the ethical debate on practicing procedures on the newly dead. AB - Emergency medicine and its academic teaching programs face an ethical dilemma surrounding the question of practicing procedures on the newly dead. For many years, procedures have been practiced on the newly dead, but few institutions have had policies addressing the practice. This article considers the ethical arguments both for and against practicing procedures on the newly dead without consent, reviews the empirical studies on the subject, and presents the positions of other professional societies, before concluding with the position of the Society for Academic Emergency Medicine (SAEM). SAEM strongly encourages all emergency medicine training programs to develop a policy and make that policy available to the institution, educators, trainees, and the public. The practice should not occur behind closed doors or on an ad hoc basis without clearly articulated guidelines. With improvements in technology, including patient simulation and virtual reality, the need for the practice may decrease, but there is no current evidence that is compelling regarding the best methods of teaching procedural skills. Given the importance of protecting trust in the profession of medicine and the existing evidence that the public would expect that consent be obtained, SAEM recommends that families be asked for consent prior to practicing procedures on the newly dead. PMID- 15347549 TI - The effect of magnesium on length of stay for pediatric sickle cell pain crisis. AB - OBJECTIVES: To determine the effect of intravenous (IV) magnesium sulfate on the length of stay (LOS) for children admitted with sickle cell pain crisis. METHODS: This was a single arm study, with historical controls. Hemoglobin SS or Hemoglobin Sbeta degrees children admitted for pain crisis were eligible. Two sequential treatment protocols of IV magnesium sulfate were added to standard inpatient therapy of IV fluids, opioids, and nonsteroidal anti-inflammatory drugs. The initial 12 children received 40 mg/kg (maximum of 1.5 g) of IV magnesium in the emergency department (ED), and eight and 16 hours later. Seven subsequent children received 40 mg/kg (maximum of 2.5 g) of IV magnesium in the ED, and every eight hours for up to four inpatient days. The main outcome measure was LOS, measured in days. Differences in LOS were analyzed between the admission with magnesium, and the two previous admissions meeting the same criteria, using a Wilcoxon signed-rank test for nonparametric paired data. RESULTS: Nineteen children were enrolled. The results of the two protocols were combined for the analysis. A shorter LOS was found on study admissions (median LOS, 3.0 days compared with 5.0 and 4.0 days for the previous two admissions; p = 0.006). CONCLUSIONS: IV magnesium appears to decrease the LOS for children with sickle cell pain crisis. PMID- 15347550 TI - Loss of paramedic availability in an urban emergency medical services system during a severe acute respiratory syndrome outbreak. AB - OBJECTIVES: To describe the loss of paramedic availability to Toronto Emergency Medical Services during a biphasic (SARS-1 and SARS-2) outbreak of severe acute respiratory syndrome (SARS). METHODS: During the SARS outbreak, a dedicated paramedic surveillance and quarantine program was developed. The authors determined the number of paramedics on quarantine each day, the type of quarantine (either home quarantine [HQ] or work quarantine [WQ]), and the development of SARS-like symptoms. RESULTS: During the SARS outbreak, there were five cases of probable SARS and three cases of suspect SARS. SARS-1 lasted 30 days, during which 234 paramedics were placed on HQ. The total number of HQ days was 1,615. During the five peak days of SARS-1, the total number of HQ days was 664. SARS-2 lasted 18 days, during which 292 paramedics were placed on either HQ or WQ, for a combined number of quarantine days of 1,637. During the five peak days of SARS-2, the combined number of quarantine days was 910. Of these, paramedics were available for duty on 708 days (78%) due to the WQ program. The primary reason for quarantine was unprotected exposure to a health care institution experiencing a SARS outbreak. Under quarantine, SARS-like symptoms developed in 68 paramedics, including cough (53 [78%]), myalgia (33 [48%]), fatigue (30 [44%]), headache (29 [43%]), fever (11 [16%]), and shortness of breath (7 [10%]). CONCLUSIONS: Paramedics were among the health care workers who developed SARS. During SARS-2, WQ optimized the number of days on which paramedics were available for duty. Many paramedics developed SARS-like symptoms without being diagnosed as having SARS. A dedicated paramedic surveillance and quarantine program provided a useful means to manage the paramedic resource during the SARS outbreak. PMID- 15347551 TI - Prevalence of elevated troponin I in end-stage renal disease patients receiving hemodialysis. AB - OBJECTIVES: Patients with end-stage renal disease (ESRD) who are receiving hemodialysis have an increased incidence of atherosclerotic heart disease. Currently, there is considerable controversy regarding the significance of elevated cardiac troponin I (cTnI) levels in patients with ESRD. The objective of this study was to examine the prevalence of elevated cTnI levels in asymptomatic patients with ESRD on hemodialysis. METHODS: This was a prospective cohort study at an urban hospital outpatient dialysis center. Serum was collected pre- and postdialysis from patients on dialysis at this center over a two-week period. Samples were batched and analyzed for cTnI using the Asxym fluorescent antibody test. An elevated cTnI level was defined as >0.8 ng/dL. RESULTS: Predialysis cTnI levels ranged from <0.1 to 0.7 ng/dL, and postdialysis cTnI levels ranged from <0.1 to 0.6 ng/dL. None of the patients had a cTnI level >0.8 ng/dL. CONCLUSIONS: cTnI levels are not falsely elevated in patients with ESRD on hemodialysis. In a patient with ESRD, myocardial injury should be suspected if his or her cTnI level is >0.8 ng/dL. PMID- 15347552 TI - Electrocardiogram interpretation training and competency assessment in emergency medicine residency programs. AB - OBJECTIVES: To determine the type of electrocardiogram (ECG) interpretation instruction in emergency medicine (EM) residency programs, the use and perceived value of teaching modalities and resources, and the methods used to assess competency of ECG interpretation. METHODS: An interactive survey instrument was posted on the Internet using SurveySuite, Inc., software and e-mailed to program directors (PDs) of all 125 Accreditation Council for Graduate Medical Education approved U.S. EM residency programs. Responses are reported in total numbers and percentages. RESULTS: Ninety-nine of 125 PDs completed the online survey (response rate, 79.2%). Emergency department instruction (99%), case-based lectures (98%), and didactic lectures (98%) were most commonly used to teach interpretation of ECGs, followed by computer-based instruction (34%) and ECG laboratory (12%). The majority of programs (53%) spent more than eight hours on formal ECG lectures per year, while 11% spent less than three hours. Observation during clinical time (99%), lecture time (76%), and hypothetical cases (57%) were the most common ways to determine competency in reading ECGs, while clinical observation and hypothetical cases were perceived as the most valuable. The most commonly used resource was personal or departmental ECG files (91%), and this had the highest perceived value. The majority of PDs were comfortable with residents' abilities to read ECGs by the third year (96%) and fourth year (91%) of residency. CONCLUSIONS: These data suggest that EM PDs believe that EM residency is adequately preparing graduates to interpret ECGs. This goal is achieved through a variety of methods. PMID- 15347553 TI - SAEM abstracts to articles: 1997 and 1999-2001. AB - OBJECTIVES: The Society for Academic Emergency Medicine (SAEM) annual meeting is the primary research conference in emergency medicine. An abstract presented at the 2000 SAEM meeting found a trend of decreasing publication rates of articles based on SAEM abstracts from 47% in 1995 to 33% in 1997. The authors wished to determine whether the publication rates of articles based on SAEM abstracts continued to decline since 1997. METHODS: A MEDLINE search of all SAEM abstracts from 1997 and 1999-2001 was conducted. The primary outcome was the publication rates of articles based on abstracts presented each year. Secondary outcomes included comparison of publication rates by oral versus poster presentation and category of research. RESULTS: Thirty-eight percent of 2,054 SAEM abstracts were published as articles by fall 2003, with 40% in 1997, 40% in 1999, 38% in 2000, and 35% in 2001. The publication rate was higher for oral presentations (50%) than poster presentations (34%). Publication rates ranged from 32% to 53% by category of research; the highest rates were in wound (53%) and cardiopulmonary resuscitation/resuscitation (51%), and the lowest rates were in administration (32%) and cardiovascular (33%). Twenty-six percent of articles were published in emergency medicine journals. The times to publication were similar across the years: 4%-7% were published in the same year of the meeting, 15%-17% one year later, 10%-11% two years later, 5% three years later, and 3%-5% after three years. CONCLUSIONS: The publication rate of articles based on SAEM abstracts has held steady at 40% in recent years when the time delay to publication is taken into account. PMID- 15347554 TI - Gender and the council of emergency medicine residency directors standardized letter of recommendation. AB - OBJECTIVES: Until 2002, the Council of Emergency Medicine Residency Directors standardized letter of recommendation (SLOR) prompted authors to predict how an applicant would rank on their match list. A ranking of guaranteed match (GM) was identified as the least common superlative response on the SLOR. That knowledge allowed precise identification of the best SLORs. The authors correlated GM with every possible author/applicant gender combination. METHODS: This was a retrospective, observational study of 835 SLORs submitted in the 1998-1999 and 1999-2000 application cycles to one emergency medicine residency program. A standardized data collection instrument was used. Author/applicant gender combinations (M/M, M/F, F/F, F/M, M/M + F/F, and M/F + F/M) were analyzed with respect to GM by chi-square test, odds ratios with 95% confidence intervals, and logistic regression. RESULTS: There was a statistically significant association between a female-authored/female-applicant SLOR and GM, with a female applicant two times more likely to get a GM from a female author than any other author/applicant gender combination (odds ratio, 2.0; 95% confidence interval = 1.1 to 3.8; p = 0.023). No other combination was significantly associated with GM. CONCLUSIONS: Female applicants to the authors' emergency residency program had a two times better chance of receiving a GM recommendation on a SLOR written by a female faculty member compared with any other possible gender combination of applicants and letter authors. Although the choice of GM has now been eliminated from the SLOR, the role of gender in relation to the SLOR merits further study. PMID- 15347555 TI - Inaccuracies on applications for emergency medicine residency training. AB - OBJECTIVES: Studies have shown erroneous claims of authorship by medical students applying for residency. Authors have hypothesized that investigation of advanced degrees, Alpha Omega Alpha (AOA) status, and peer-reviewed publications all show important rates of inaccuracy. METHODS: A retrospective review of all applicants offered an interview for the authors' emergency medicine (EM) residency (entering class of 2002), excluding foreign medical graduates and current residents, was conducted. After verifying peer-reviewed publications by MEDLINE search and journal review, errors were tabulated as follows: reference not found, not referenced as an abstract, incorrect author list, or clerical error. AOA status was verified by the AOA organization. Advanced degrees were verified by the awarding institutions. RESULTS: Of 194 applications screened (58.3% of applications), 21 (10.8%) were excluded (9 foreign medical graduates, 12 current residents). Multiple inaccuracies on a single application were counted separately. Of the 173 remaining applications, 23 (13.3%; 95% confidence interval [95% CI] = 8.8% to 19.5%) had at least one misrepresentation and seven of 173 (4.0%; 95% CI = 1.8% to 8.5%) had two or more. Authorship of at least one peer reviewed article was claimed by 47 of 173 (27.2%), with ten of 47 (21.3%; 95% CI = 11.2% to 36.1%) having one inaccuracy and six of 47 (12.8%, 95% CI = 5.3% to 26.4%) having two or more. AOA membership was claimed by 14 applicants (8.1%), but five claims (35.7%, 95% CI = 14.0% to 64.4%) were inaccurate. Advanced degrees were claimed by 15 (8.7%); four (26.7%, 95% CI = 8.9% to 55.2%) were in error. CONCLUSIONS: Applications for EM residency contain frequent inaccuracies in publications listed, AOA status, and advanced degrees. Careful review of applications is necessary to ensure appropriate credit is given for claims of these types. PMID- 15347556 TI - Ethics in the emergency department. PMID- 15347557 TI - Stomatal conductance of lettuce grown under or exposed to different light qualities. AB - BACKGROUND AND AIMS: The objective of this research was to examine the effects of differences in light spectrum on the stomatal conductance (Gs) and dry matter production of lettuce plants grown under a day/night cycle with different spectra, and also the effects on Gs of short-term exposure to different spectra. METHODS: Lettuce (Lactuca sativa) plants were grown with 6 h dark and 18 h light under four different spectra, red-blue (RB), red-blue-green (RBG), green (GF) and white (CWF), and Gs and plant growth were measured. KEY RESULTS AND CONCLUSIONS: Conductance of plants grown for 23 d under CWF rose rapidly on illumination to a maximum in the middle of the light period, then decreased again before the dark period when it was minimal. However, the maximum was smaller in plants grown under RB, RGB and GF. This demonstrates that spectral quality during growth affects the diurnal pattern of stomatal conductance. Although Gs was smaller in plants grown under RGB than CWF, dry mass accumulation was greater, suggesting that Gs did not limit carbon assimilation under these spectral conditions. Temporarily changing the spectral quality of the plants grown for 23 d under CWF, affected stomatal responses reversibly, confirming studies on epidermal strips. This study provides new information showing that Gs is responsive to spectral quality during growth and, in the short-term, is not directly coupled to dry matter accumulation. PMID- 15347558 TI - Endothelin-1 inhibits mucin secretion from ovine airway epithelial goblet cells. AB - Mucus hypersecretion is a feature of several respiratory diseases and frequently leads to obstruction of small airways where the principal source of mucous glycoproteins (mucins), the major macromolecular constituents of mucus, are goblet cells. Hence, inhibition of mucin secretion from these cells may be clinically beneficial. In this study, we have developed a lectin-based assay for mucin secretion from ovine airway goblet cells and used this assay to investigate the regulation of these cells by endothelin (ET)-1. ET-1 inhibited baseline mucin secretion (maximum inhibition: 60.3 +/- 4.2%, 50% inhibitory concentration: 0.8 +/- 0.17 nM). This response was abolished by the ET(A) antagonist, BQ-123 (1 muM), but not by the ET(B) antagonist, BQ-788 (1 muM). ET-1 (1 muM) did not affect mucin secretion stimulated by ATP (100 muM) but secretion in response to ATP (10 muM) was inhibited by 63.3 +/- 11.8%. This response could be eliminated by BQ-123, but not by BQ-788. Radioligand binding and immunohistochemistry indicated the expression of both ET(A)- and ET(B)-receptors on the epithelium. In summary, ET-1, acting via ET(A)-receptors, inhibits baseline and ATP-stimulated mucin secretion from ovine airway goblet cells. This represents the first report of a physiologic mechanism for inhibiting airway goblet cell mucin secretion; an understanding of this mechanism may provide opportunities for the treatment of obstructive airways disease. PMID- 15347559 TI - Immune response to Mycoplasma pulmonis in nasal mucosa is modulated by the normal microbiota. AB - The impact of commensal bacteria on lymphocyte responses in the upper airways was studied in rat nasal mucosa after infection with the pathogen Mycoplasma pulmonis. Phenotyping was performed in situ by paired immunofluorescence staining in germ-free (GF) and conventional (CV) rats before and 3 wk after the monoinfection. Intraepithelial lymphocytes had expanded significantly in GF (P = 0.02) but not in CV rats. Furthermore, a striking proportional increase of T-cell receptor (TCR)alphabeta(+)CD4(+) cells was observed both in the lamina propria and epithelium of GF (P < 0.01) but not of CV rats. Notably, in contrast to the pre-infection state, both mucosal compartments showed a percentage of TCRalphabeta(+)CD4(+) cells that was significantly higher in GF (P = 0.03-P < 0.01) than in CV rats after the monoinfection. In parallel, both compartments displayed a percentage of TCRalphabeta(+) CD8(+) cells that was decreased in GF (P < 0.01) but not in CV rats. The small fraction of TCRgammadelta(+) T cells observed (< 5%) did not change quantitatively or phenotypically after infection. The size of organized nose-associated lymphoid tissue was, on average, increased 5.2-fold in GF rats versus 2.6-fold in CV rats. Collectively, our results demonstrated that the normal microbiota modulated markedly the nasal immune response elicited by monoinfection with M. pulmonis. PMID- 15347560 TI - Opposing effects of 60% oxygen and neutrophil influx on alveologenesis in the neonatal rat. AB - The lungs of newborn rats exposed to 60% oxygen for 14 days develop an injury that shares morphologic similarities to human bronchopulmonary dysplasia (BPD). Neutrophil influx into the lung, as part of an inflammatory response, may play a pivotal role in the development of BPD. A neutrophil chemokine, cytokine-induced neutrophil chemoattractant-1, which signals through the neutrophil CXC chemokine receptor-2, is increased in the lung tissue of newborn rats exposed to 60% oxygen. The purpose of this study was to explore the role of neutrophils in the rat model of BPD by inhibiting neutrophil influx using SB265610, a selective CXC chemokine receptor-2 antagonist. SB265610, administered to 60% oxygen-exposed newborn rats from birth to 14 days, completely inhibited neutrophil influx. It also attenuated increased production of reactive oxygen species in newborn rat lung tissue after exposure to 60% oxygen for 4 days. Lung morphometric analysis revealed that 60% oxygen for 14 days, when accompanied by treatment with SB265610 to prevent neutrophil accumulation, increased alveolar formation over that seen in newborn rats exposed to air. These data suggest that exposure of the neonatal lung to moderate hyperoxia may enhance postnatal lung growth, provided postnatal pulmonary inflammation is suppressed. PMID- 15347561 TI - A case control etiologic study of sarcoidosis: environmental and occupational risk factors. AB - Past research suggests that environmental factors may be associated with sarcoidosis risk. We conducted a case control study to test a priori hypotheses that environmental and occupational exposures are associated with sarcoidosis. Ten centers recruited 706 newly diagnosed patients with sarcoidosis and an equal number of age-, race-, and sex-matched control subjects. Interviewers administered questionnaires containing questions regarding occupational and nonoccupational exposures that we assessed in univariable and multivariable analyses. We observed positive associations between sarcoidosis and specific occupations (e.g., agricultural employment, odds ratio [OR] 1.46, confidence interval [CI] 1.13-1.89), exposures (e.g., insecticides at work, OR 1.52, CI 1.14 2.04, and work environments with mold/mildew exposures [environments with possible exposures to microbial bioaerosols], OR 1.61, CI 1.13-2.31). A history of ever smoking cigarettes was less frequent among cases than control subjects (OR 0.62, CI 0.50-0.77). In multivariable modeling, we observed elevated ORs for work in areas with musty odors (OR 1.62, CI 1.24-2.11) and with occupational exposure to insecticides (OR 1.61, CI 1.13-2.28), and a decreased OR related to ever smoking cigarettes (OR 0.65, CI 0.51-0.82). The study did not identify a single, predominant cause of sarcoidosis. We identified several exposures associated with sarcoidosis risk, including insecticides, agricultural employment, and microbial bioaerosols. PMID- 15347562 TI - Prevalence of sleep-disordered breathing in middle-aged Korean men and women. AB - With many epidemiologic studies made to establish the prevalence of sleep disordered breathing (SDB) and obstructive sleep apnea syndrome (OSAS) in Western countries, no such data have been reported in Korea. The purpose of this study was to examine the prevalence of SDB and OSAS, and their related factors in Korean adults aged 40-69 years. Among the total of 5,020 participants at the baseline examination of the Korean Health and Genome Study, a random sample of 457 men and women was studied with employment of overnight full polysomnography to determine the prevalence of SDB and OSAS. The prevalence of SDB (apnea hypopnea index > or = 5) was 27% and 16% in men and women, respectively. When OSAS was defined by an apnea-hypopnea index > or = 5 plus excessive daytime sleepiness, its prevalence was 4.5% in men and 3.2% in women. Logistic regression analyses showed that sex, body mass index, and hypertension were closely associated with the risk of SDB. Our findings show that SDB is a common problem in the Korean adult population. Understanding and treatment of SDB may be essential in terms of intervention to reduce the risk of related medical problems. PMID- 15347563 TI - Genome-wide linkage of forced mid-expiratory flow in chronic obstructive pulmonary disease. AB - Familial aggregation of forced expiratory flow during the middle half of the FVC (FEF(25-75%)) and FEF(25-75%)/FVC has been observed in the Boston Early-Onset Chronic Obstructive Pulmonary Disease Study, but linkage results have not been reported for these phenotypes. An autosomal whole genome-wide linkage scan was performed in 72 pedigrees ascertained through a proband with severe, early-onset chronic obstructive pulmonary disease, and linkage analyses of FEF(25-75%) and FEF(25-75%)/FVC were performed using Sequential Oligogenic Linkage Analysis Routines. There was suggestive evidence for linkage of FEF(25-75%)/FVC with chromosome 2 (LOD 2.60 at 216 cM). In a smokers-only analysis, evidence for linkage was observed for postbronchodilator FEF(25-75%) with chromosome 12 (LOD 5.03 at 35 cM) and chromosomes 2 and 12 for FEF(25-75%)/FVC (LOD 4.12 at 221 cM and LOD 3.46 at 35 cM, respectively); in the smokers-only model, evidence for linkage also was robust for FEV(1)/FVC on chromosome 2 (LOD 4.13 at 229 cM) and FEV(1) on chromosome 12 (LOD 3.26 at 36 cM). Our analyses provide evidence for linkage of FEF(25-75%) and FEF(25-75%)/FVC on chromosomes 2q and 12p. LOD scores of greater than two were also observed for chromosomes 16, 20, and 22 with the smokers-only analysis, which may suggest gene-by-smoking interactions in these regions. PMID- 15347564 TI - Automatic control of the inspired oxygen fraction in preterm infants: a randomized crossover trial. AB - In preterm infants receiving supplemental oxygen, manual control of the inspired oxygen fraction is often time-consuming and inappropriate. We developed a system for automatic oxygen control and hypothesized that this system is more effective than routine manual oxygen control in maintaining target arterial oxygen saturation levels. We performed a randomized controlled crossover clinical trial in 12 preterm infants receiving nasal continuous positive airway pressure and supplemental oxygen. Periods with automatic and routine manual oxygen control were compared with periods of optimal control by a fully dedicated person. The median (range) percentage of time with arterial oxygen saturation levels within target range (87-96%) was 81.7% (39.0-99.8) for routine manual oxygen control, 91.0% (41.4-99.3) for optimal control, and 90.5% (59.0-99.4) for automatic control (ANOVA: p = 0.01). Pairwise post hoc comparisons revealed a statistically significant difference between automatic and routine manual oxygen control (Dunnett's test: p = 0.02). The frequency of manual oxygen adjustments was lowest in automatic control (Friedman's test: p < 0.001). Automatic oxygen control may optimize oxygen administration to preterm infants receiving nasal continuous positive airway pressure and reduce nursing time spent with oxygen control. PMID- 15347565 TI - Endothelial alterations during inhaled NO in lambs with pulmonary hypertension: implications for rebound hypertension. AB - Clinically significant increases in pulmonary vascular resistance (PVR) have been noted upon acute withdrawal of inhaled nitric oxide (iNO). Previous studies in the normal pulmonary circulation demonstrate that iNO increases endothelin-1 (ET 1) levels and decreases endogenous nitric oxide synthase (NOS) activity, implicating an endothelial etiology for the increase in resistance upon iNO withdrawal. However, the effect of iNO on endogenous endothelial function in the clinically relevant pulmonary hypertensive circulation is unknown. The objective of this study was to determine the effects of iNO on endogenous NO-cGMP and ET-1 signaling in lambs with preexisting pulmonary hypertension secondary to increased pulmonary blood flow. Eight fetal lambs underwent in utero placement of an aortopulmonary vascular graft (shunt lambs). After delivery (4 wk), the shunt lambs were mechanically ventilated with iNO (40 ppm) for 24 h. After 24 h of inhaled NO, plasma ET-1 levels increased by 34.8% independently of changes in protein levels (P < 0.05). Contrary to findings in normal lambs, total NOS activity did not decrease during iNO. In fact, Western blot analysis demonstrated that tissue endothelial NOS protein levels decreased by 43% such that NOS activity relative to protein levels actually increased during iNO (P < 0.05). In addition, the beta-subunit of soluble guanylate cyclase decreased by 70%, whereas phosphodiesterase 5 levels were unchanged (P < 0.05). Withdrawal of iNO was associated with an acute increase in PVR, which exceeded baseline PVR by 45%, and a decrease in cGMP concentrations to levels that were below baseline. These data suggest that the endothelial response to iNO and the potential mechanisms of rebound pulmonary hypertension are dependent upon the underlying pulmonary vasculature. PMID- 15347566 TI - Tumor necrosis factor-alpha-induced TRPC1 expression amplifies store-operated Ca2+ influx and endothelial permeability. AB - We determined the effects of TNF-alpha on the expression of transient receptor potential channel (TRPC) homologues in human vascular endothelial cells and the consequences of TRPC expression on the endothelial permeability response. We observed that TNF-alpha exposure increased TRPC1 expression without significantly altering expression of other TRPC isoforms in human pulmonary artery endothelial cells (HPAEC). Because TRPC1 belongs to the store-operated cation channel family, we measured the Ca(2+) store depletion-mediated Ca(2+) influx in response to thrombin exposure. We observed that thrombin-induced Ca(2+) influx in TNF-alpha stimulated HPAEC was twofold greater than in control cells. To address the relationship between store-operated Ca(2+) influx and TRPC1 expression, we overexpressed TRPC1 by three- to fourfold in the human dermal microvascular endothelial cell line (HMEC) using the TRPC1 cDNA. Thrombin-induced store Ca(2+) depletion in these cells caused approximately twofold greater increase in Ca(2+) influx than in control cells. Furthermore, the inositol 1,4,5-trisphosphate sensitive store-operated cationic current was increased greater than twofold in TRPC1-transfected cells compared with control. To address the role of Ca(2+) influx via TRPC1 in signaling endothelial permeability, we measured actin-stress fiber formation and transendothelial monolayer electrical resistance (TER) in the TRPC1 cDNA-transfected HMEC and TNF-alpha-challenged HPAEC. Both thrombin-induced actin-stress fiber formation and a decrease in TER were augmented in TRPC1 overexpressing HMEC compared with control cells. TNF-alpha-induced increased TRPC1 expression in HPAEC also resulted in marked endothelial barrier dysfunction in response to thrombin. These findings indicate the expression level of TRPC1 in endothelial cells is a critical determinant of Ca(2+) influx and signaling of the increase in endothelial permeability. PMID- 15347567 TI - Surfactant phospholipid changes after antigen challenge: a role for phosphatidylglycerol in dysfunction. AB - In asthma, inflammation-mediated surfactant dysfunction contributes to increased airway resistance, but the mechanisms for dysfunction are not understood. To test mechanisms that alter surfactant function, atopic asthmatics underwent endobronchial antigen challenge and bronchoalveolar lavage (BAL). BAL fluids were sequentially separated into cells, surfactant, and supernatant, and multiple end points were analyzed. Each end point's unique relationship to surfactant dysfunction was determined. Our results demonstrate that minimum surface tension (gamma(min)) of surfactant after antigen challenge was significantly increased with a spectrum of responses that included dysfunction in 6 of 13 asthmatics. Antigen challenge significantly altered the partitioning of surfactant phospholipid measured as a decreased ratio of large surfactant aggregates (LA) to small surfactant aggregates (SA), LA/SA ratio. Phosphatidylglycerol (PG) was significantly reduced in the LA of the dysfunctional asthmatic BALs. There was a corresponding significant increase in the ratio of phosphatidylcholine to PG, which strongly correlated with both increased gamma(min) and decreased LA/SA. Altered surfactant phospholipid properties correlated with surfactant dysfunction as well or better than either increased eosinophils or protein. Secretory phospholipase activity, measured in vitro, increased after antigen challenge and may explain the decrease in surfactant PG. In summary, alteration of phospholipids, particularly depletion of PG, in the LA of surfactant may be an important mechanism in asthma-associated surfactant dysfunction. PMID- 15347568 TI - Keratinocyte growth factor induces Akt kinase activity and inhibits Fas-mediated apoptosis in A549 lung epithelial cells. AB - Acute respiratory distress syndrome (ARDS) is a syndrome characterized by the rapid influx of protein-rich edema fluid into the air spaces. The magnitude of alveolar epithelial cell injury is a key determinant of disease severity and an important predictor of patient outcome. The alveolar epithelium is positioned at the interface of the host response in the initiation, progression, and recovery phase of the disease. Keratinocyte growth factor (KGF) is a potent survival factor unique to the epithelium that promotes lung epithelial cell survival, accelerates wound closure, and reduces fibrosis. We therefore hypothesized that KGF preserves lung function by inhibiting apoptosis through activation of a signal transduction pathway responsible for cell survival. To test this hypothesis we determined that KGF inhibits death following Fas activation, a relevant apoptosis pathway, and then determined that cell survival is mediated through activation of the phosphatidylinositol 3'-kinase (PI3K)/Akt kinase signal transduction pathway. We found that KGF induces a dose- and time-dependent increase in Akt kinase activity and that, as expected, activation of Akt via KGF is PI3K dependent. KGF inhibited Fas-induced apoptosis as measured by a reduction in apoptotic cells and caspase-3 activity. This investigation supports our original hypothesis that KGF protects the lung epithelium by inhibiting apoptosis and that protection occurs through activation of PI3K/Akt-mediated cell survival pathway. Our results are in agreement with other reports that identify the PI3K/Akt axis as a key intracellular pathway in the lung epithelium that may serve as a therapeutic target to preserve epithelial integrity during inflammation. PMID- 15347569 TI - Developmental regulation of claudin localization by fetal alveolar epithelial cells. AB - Tight junction proteins in the claudin family regulate epithelial barrier function. We examined claudin expression by human fetal lung (HFL) alveolar epithelial cells cultured in medium containing dexamethasone, 8-bromo-cAMP, and isobutylmethylxanthanine (DCI), which promotes alveolar epithelial cell differentiation to a type II phenotype. At the protein level, HFL cells expressed claudin-1, claudin-3, claudin-4, claudin-5, claudin-7, and claudin-18, where levels of expression varied with culture conditions. DCI-treated differentiated HFL cells cultured on permeable supports formed tight transepithelial barriers, with transepithelial resistance (TER) >1,700 ohm/cm(2). In contrast, HFL cells cultured in control medium without DCI did not form tight barriers (TER <250 ohm/cm(2)). Consistent with this difference in barrier function, claudins expressed by HFL cells cultured in DCI medium were tightly localized to the plasma membrane; however, claudins expressed by HFL cells cultured in control medium accumulated in an intracellular compartment and showed discontinuities in claudin plasma membrane localization. In contrast to claudins, localization of other tight junction proteins, zonula occludens (ZO)-1, ZO-2, and occludin, was not sensitive to HFL cell phenotype. Intracellular claudins expressed by undifferentiated HFL cells were localized to a compartment containing early endosome antigen-1, and treatment of HFL cells with the endocytosis inhibitor monodansylcadaverine increased barrier function. This suggests that during differentiation to a type II cell phenotype, fetal alveolar epithelial cells use differential claudin expression and localization to the plasma membrane to help regulate tight junction permeability. PMID- 15347570 TI - ProViz: protein interaction visualization and exploration. AB - ProViz is a tool for the visualization of protein-protein interaction networks, developed by the IntAct European project. It provides facilities for navigating in large graphs and exploring biologically relevant features, and adopts emerging standards such as GO and PSI-MI. AVAILABILITY: ProViz is available under the GPL and may be freely downloaded. Source code and binaries are available at http://cbi.labri.fr/eng/proviz.htm CONTACT: david.sherman@labri.fr PMID- 15347571 TI - BarleyExpress: a web-based submission tool for enriched microarray database annotations. AB - BarleyExpress is a web-based microarray experiment data submission tool for BarleyBase, a public data resource of Affymetrix GeneChip data for plants. BarleyExpress uses the Plant Ontology vocabularies and enhances the MIAME guidelines to standardize the annotation of microarray gene expression experiments. In addition, BarleyExpress provides explicit support for factorial experiment design and template loading methods to ease the submission process for large experiments. AVAILABILITY: http://barleybase.org SUPPLEMENTARY INFORMATION: BarleyExpress Users Manual. PMID- 15347572 TI - A more efficient search strategy for aging genes based on connectivity. AB - MOTIVATION: Many aging genes have been found from unbiased screens in model organisms. Genetic interventions promoting longevity are usually quantitative, while in many other biological fields (e.g. development) null mutations alone have been very informative. Therefore, in the case of aging the task is larger and the need for a more efficient genetic search strategy is especially strong. RESULTS: The topology of genetic and metabolic networks is organized according to a scale-free distribution, in which hubs with large numbers of links are present. We have developed a computational model of aging genes as the hubs of biological networks. The computational model shows that, after generalized damage, the function of a network with scale-free topology can be significantly restored by a limited intervention on the hubs. Analyses of data on aging genes and biological networks support the applicability of the model to biological aging. The model also might explain several of the properties of aging genes, including the high degree of conservation across different species. The model suggests that aging genes tend to have a higher number of connections and therefore supports a strategy, based on connectivity, for prioritizing what might otherwise be a random search for aging genes. PMID- 15347573 TI - SNPbox: a modular software package for large-scale primer design. AB - We developed a modular software package SNPbox that automates and standardizes the generation of PCR primers and is used in the strategy for constructing single nucleotide polymorphisms (SNPs) maps. In this strategy, the focus of primer design can be either on the validation of annotated public SNPs or on the SNP discovery in exon regions or extended genomic regions, both by resequencing. SNPbox relies on Primer3 for the primer design and combines this program with other publicly available software tools such as BLAST, Spidey and RepeatMasker, and newly developed algorithms. Primer conditions were chosen such that PCR amplifications are uniform for each PCR amplicon facilitating the use of high throughput genetic platforms. SNPbox can also be used for the design of primer sets for mutation analysis, STR marker genotyping and microarray oligo design. Of the 2500 primer sets designed by SNPbox, 95% successfully amplified genomic DNA under uniform PCR conditions. AVAILABILITY: The software is available from the authors upon request. SUPPLEMENTARY INFORMATION: SNPbox_supplement. PMID- 15347574 TI - Detecting overlapping coding sequences with pairwise alignments. AB - MOTIVATION: Overlapping gene coding sequences (CDSs) are particularly common in viruses but also occur in more complex genomes. Detecting such genes with conventional gene-finding algorithms can be difficult for several reasons. If an overlapping CDS is on the same read-strand as a known CDS, then there may not be a distinct promoter or mRNA. Furthermore, the constraints imposed by double coding can result in atypical codon biases. However, these same constraints lead to particular mutation patterns that may be detectable in sequence alignments. RESULTS: In this paper, we investigate several statistics for detecting double coding sequences with pairwise alignments--including a new maximum-likelihood method. We also develop a model for double-coding sequence evolution. Using simulated sequences generated with the model, we characterize the distribution of each statistic as a function of sequence composition, length, divergence time and double-coding frame. Using these results, we develop several algorithms for detecting overlapping CDSs. The algorithms were tested on known overlapping CDSs and other overlapping open reading frames (ORFs) in the hepatitis B virus (HBV), Escherichia coli and Salmonella typhimurium genomes. The algorithms should prove useful for detecting novel overlapping genes--especially short coding ORFs in viruses. AVAILABILITY: Programs may be obtained from the authors. SUPPLEMENTARY INFORMATION: http://biochem.otago.ac.nz/double.html. PMID- 15347575 TI - Evaluating putative chimeric sequences from PCR-amplified products. AB - MOTIVATION: PCR amplification of highly homologous genes from complex DNA mixtures is known to generate a significant proportion of chimeric sequences. Ribosomal RNA genes are used for microbial species detection and identification in natural environments, and current assessments of microbial diversity are based on these sequences. Thus, chimeric sequences could lead to the discovery of non existent microbial species and false diversity estimates. METHODS: In essence, our only source of information to decide if a sequence is chimeric or not is to compare it with known, non-chimeric sequences. Putative chimeric sequences were analyzed from sequence fragments of selected length (referred to as words) by comparing nucleotides at corresponding positions. Distances for each word between reference sequences (closely related to the tested sequence) were compared to the differences introduced by the tested sequence. The proposed strategy considers the actual variability existing in different regions throughout the analyzed sequences. The result is an efficient strategy for the evaluation of putative chimeric sequences. AVAILABILITY: A program computing the above procedure, Chimera and Cross-Over Detection and Evaluation (Ccode), is available at http://www.irnase.csic.es/users/jmgrau/index.html and http://www.rtphc.csic.es/download.html. PMID- 15347576 TI - easyLINKAGE: a PERL script for easy and automated two-/multi-point linkage analyses. AB - We have generated the program easyLINKAGE that combines automated setup and performance of linkage analyses and simulation under an easy to handle graphical user interface for Microsoft Windows 2000/XP and standard UNIX systems. The program package supports two-point linkage analyses (FastLink v4.1 and SPLink v1.09), multi-point linkage analyses [GENEHUNTER v2.1, GENEHUNTER-PLUS with the emendation by Kong and Cox v1.2 (allele sharing modelling)] and the simulation package SLINK v2.65, and provides genome-wide as well as chromosomal postscript plots of LOD scores, NPL scores, P-values and other parameters. AVAILABILITY: http://www.uni-wuerzburg.de/nephrologie/molecular_genetics/molecular_genetics.htm SUPPLEMENTARY INFORMATION: Supplementary information is available on the website. PMID- 15347577 TI - Tracker: continuous HMMER and BLAST searching. AB - SUMMARY: Tracker is a web-based email alert system for monitoring protein database searches using HMMER and Blast-P, nucleotide searches using Blast-N and literature searches of the PubMed database. Users submit searches via a web-based interface. Searches are saved and run against updated databases to alert users about new information. If there are new results from the saved searches, users will be notified by email and will then be able to access results and link to additional information on the NCBI website. Tracker supports Boolean AND/OR operations on HMMER and BLASTP result sets to allow users to broaden or narrow protein searches. AVAILABILITY: The server is located at http://jay.bioinformatics.ku.edu/tracker/index.html. A distribution package including detailed installation procedure is freely available from http://jay.bioinformatics.ku.edu/download/tracker/. PMID- 15347578 TI - The SSEA server for protein secondary structure alignment. AB - SUMMARY: We present a web server that computes alignments of protein secondary structures. The server supports both performing pairwise alignments and searching a secondary structure against a library of domain folds. It can calculate global and local secondary structure element alignments. A combination of local and global alignment steps can be used to search for domains inside the query sequence or help in the discrimination of novel folds. Both the SCOP and PDB fold libraries, clustered at 95 and 40% sequence identity, are available for alignment. AVAILABILITY: The web server interface is freely accessible to academic users at http://protein.cribi.unipd.it/ssea/. The executable version and benchmarking data are available from the same web page. PMID- 15347579 TI - Genome Properties: a system for the investigation of prokaryotic genetic content for microbiology, genome annotation and comparative genomics. AB - MOTIVATION: The presence or absence of metabolic pathways and structures provide a context that makes protein annotation far more reliable. Compiling such information across microbial genomes improves the functional classification of proteins and provides a valuable resource for comparative genomics. RESULTS: We have created a Genome Properties system to present key aspects of prokaryotic biology using standardized computational methods and controlled vocabularies. Properties reflect gene content, phenotype, phylogeny and computational analyses. The results of searches using hidden Markov models allow many properties to be deduced automatically, especially for families of proteins (equivalogs) conserved in function since their last common ancestor. Additional properties are derived from curation, published reports and other forms of evidence. Genome Properties system was applied to 156 complete prokaryotic genomes, and is easily mined to find differences between species, correlations between metabolic features and families of uncharacterized proteins, or relationships among properties. AVAILABILITY: Genome Properties can be found at http://www.tigr.org/Genome_Properties SUPPLEMENTARY INFORMATION: http://www.tigr.org/tigr-scripts/CMR2/genome_properties_references.spl. PMID- 15347580 TI - Country of training and ethnic origin of UK doctors: database and survey studies. AB - OBJECTIVES: To report on the country of training and ethnicity of consultants in different specialties in the NHS, on trends in intake to UK medical schools by ethnicity, and on the specialty choices made by UK medical graduates in different ethnic groups. DESIGN: Analysis of official databases of consultants and of students accepted to study medicine; survey data about career choices made by newly qualified doctors. SETTING AND SUBJECTS: England and Wales (consultants), United Kingdom (students and newly qualified doctors). RESULTS: Of consultants appointed before 1992, 15% had trained abroad; of those appointed in 1992-2001, 24% had trained abroad. The percentage of consultants who had trained abroad and were non-white was significantly high, compared with their overall percentage among consultants, in geriatric medicine, genitourinary medicine, paediatrics, old age psychiatry, and learning disability. UK trained non-white doctors had specialty destinations similar to those of UK trained white doctors. The percentage of UK medical graduates who are non-white has increased substantially from about 2% in 1974 and will approach 30% by 2005. White men now comprise little more than a quarter of all UK medical students. White and non-white UK graduates make similar choices of specialty. CONCLUSIONS: Specialist medical practice in the NHS has been heavily dependent on doctors who have trained abroad, particularly in specialties where posts have been hard to fill. By contrast, UK trained doctors from ethnic minorities are not over-represented in the less popular specialties. Ethnic minorities are well represented in UK medical school intakes; and white men, but not white women, are now substantially under-represented. PMID- 15347582 TI - A statistical thermodynamic model applied to experimental AFM population and location data is able to quantify DNA-histone binding strength and internucleosomal interaction differences between acetylated and unacetylated nucleosomal arrays. AB - Imaging of nucleosomal arrays by atomic force microscopy allows a determination of the exact statistical distributions for the numbers of nucleosomes per array and the locations of nucleosomes on the arrays. This precision makes such data an excellent reference for testing models of nucleosome occupation on multisite DNA templates. The approach presented here uses a simple statistical thermodynamic model to calculate theoretical population and positional distributions and compares them to experimental distributions previously determined for 5S rDNA nucleosomal arrays (208-12,172-12). The model considers the possible locations of nucleosomes on the template, and takes as principal parameters an average free energy of interaction between histone octamers and DNA, and an average wrapping length of DNA around the octamers. Analysis of positional statistics shows that it is possible to consider interactions between nucleosomes and positioning effects as perturbations on a random positioning noninteracting model. Analysis of the population statistics is used to determine histone-DNA association constants and to test for differences in the free energies of nucleosome formation with different types of histone octamers, namely acetylated or unacetylated, and different DNA templates, namely 172-12 or 208-12 5S rDNA multisite templates. The results show that the two template DNAs bind histones with similar affinities but histone acetylation weakens the association of histones with both templates. Analysis of locational statistics is used to determine the strength of specific nucleosome positioning tendencies by the DNA templates, and the strength of the interactions between neighboring nucleosomes. The results show only weak positioning tendencies and that unacetylated nucleosomes interact much more strongly with one another than acetylated nucleosomes; in fact acetylation appears to induce a small anticooperative occupation effect between neighboring nucleosomes. PMID- 15347581 TI - A mathematical treatment of integrated Ca dynamics within the ventricular myocyte. AB - We have developed a detailed mathematical model for Ca2+ handling and ionic currents in the rabbit ventricular myocyte. The objective was to develop a model that: 1), accurately reflects Ca-dependent Ca release; 2), uses realistic parameters, particularly those that concern Ca transport from the cytosol; 3), comes to steady state; 4), simulates basic excitation-contraction coupling phenomena; and 5), runs on a normal desktop computer. The model includes the following novel features: 1), the addition of a subsarcolemmal compartment to the other two commonly formulated cytosolic compartments (junctional and bulk) because ion channels in the membrane sense ion concentrations that differ from bulk; 2), the use of realistic cytosolic Ca buffering parameters; 3), a reversible sarcoplasmic reticulum (SR) Ca pump; 4), a scheme for Na-Ca exchange transport that is [Na]i dependent and allosterically regulated by [Ca]i; and 5), a practical model of SR Ca release including both inactivation/adaptation and SR Ca load dependence. The data describe normal electrical activity and Ca handling characteristics of the cardiac myocyte and the SR Ca load dependence of these processes. The model includes a realistic balance of Ca removal mechanisms (e.g., SR Ca pump versus Na-Ca exchange), and the phenomena of rest decay and frequency dependent inotropy. A particular emphasis is placed upon reproducing the nonlinear dependence of gain and fractional SR Ca release upon SR Ca load. We conclude that this model is more robust than many previously existing models and reproduces many experimental results using parameters based largely on experimental measurements in myocytes. PMID- 15347583 TI - Pressure denaturation of staphylococcal nuclease studied by neutron small-angle scattering and molecular simulation. AB - We studied the pressure-induced folding/unfolding transition of staphylococcal nuclease (SN) over a pressure range of approximately 1-3 kilobars at 25 degrees C by small-angle neutron scattering and molecular dynamics simulations. We find that applying pressure leads to a twofold increase in the radius of gyration derived from the small-angle neutron scattering spectra, and P(r), the pair distance distribution function, broadens and shows a transition from a unimodal to a bimodal distribution as the protein unfolds. The results indicate that the globular structure of SN is retained across the folding/unfolding transition although this structure is less compact and elongated relative to the native structure. Pressure-induced unfolding is initiated in the molecular dynamics simulations by inserting water molecules into the protein interior and applying pressure. The P(r) calculated from these simulations likewise broadens and shows a similar unimodal-to-bimodal transition with increasing pressure. The simulations also reveal that the bimodal P(r) for the pressure-unfolded state arises as the protein expands and forms two subdomains that effectively diffuse apart during initial stages of unfolding. Hydrophobic contact maps derived from the simulations show that water insertions into the protein interior and the application of pressure together destabilize hydrophobic contacts between these two subdomains. The findings support a mechanism for the pressure-induced unfolding of SN in which water penetration into the hydrophobic core plays a central role. PMID- 15347584 TI - Calcium and glycolysis mediate multiple bursting modes in pancreatic islets. AB - Pancreatic islets of Langerhans produce bursts of electrical activity when exposed to stimulatory glucose levels. These bursts often have a regular repeating pattern, with a period of 10-60 s. In some cases, however, the bursts are episodic, clustered into bursts of bursts, which we call compound bursting. Consistent with this are recordings of free Ca2+ concentration, oxygen consumption, mitochondrial membrane potential, and intraislet glucose levels that exhibit very slow oscillations, with faster oscillations superimposed. We describe a new mathematical model of the pancreatic beta-cell that can account for these multimodal patterns. The model includes the feedback of cytosolic Ca2+ onto ion channels that can account for bursting, and a metabolic subsystem that is capable of producing slow oscillations driven by oscillations in glycolysis. This slow rhythm is responsible for the slow mode of compound bursting in the model. We also show that it is possible for glycolytic oscillations alone to drive a very slow form of bursting, which we call "glycolytic bursting." Finally, the model predicts that there is bistability between stationary and oscillatory glycolysis for a range of parameter values. We provide experimental support for this model prediction. Overall, the model can account for a diversity of islet behaviors described in the literature over the past 20 years. PMID- 15347585 TI - Single molecule observation of liposome-bilayer fusion thermally induced by soluble N-ethyl maleimide sensitive-factor attachment protein receptors (SNAREs). AB - A single molecule fluorescence assay is presented for studying the mechanism of soluble N-ethyl maleimide sensitive-factor attachment protein receptors (SNAREs) mediated liposome fusion to supported lipid bilayers. The three neuronal SNAREs syntaxin-1A, synaptobrevin-II (VAMP), and SNAP-25A were expressed separately, and various dye-labeled combinations of the SNAREs were tested for their ability to dock liposomes and induce fusion. Syntaxin and synaptobrevin in opposing membranes were both necessary and sufficient to dock liposomes to supported bilayers and to induce thermally activated fusion. As little as one SNARE interaction was sufficient for liposome docking. Fusion of docked liposomes with the supported bilayer was monitored by the dequenching of soluble fluorophores entrapped within the liposomes. Fusion was stimulated by illumination with laser light, and the fusion probability was enhanced by raising the ambient temperature from 22 to 37 degrees C, suggesting a thermally activated process. Surprisingly, SNAP-25 had little effect on docking efficiency or the probability of thermally induced fusion. Interprotein fluorescence resonance energy transfer experiments suggest the presence of other conformational states of the syntaxin*synaptobrevin interaction in addition to those observed in the crystal structure of the SNARE complex. Furthermore, although SNARE complexes involved in liposome docking preferentially assemble into a parallel configuration, both parallel and antiparallel configurations were observed. PMID- 15347586 TI - Distinct effects on Ca2+ handling caused by malignant hyperthermia and central core disease mutations in RyR1. AB - Malignant hyperthermia (MH) and central core disease (CCD) are disorders of skeletal muscle Ca2+ homeostasis that are linked to mutations in the type 1 ryanodine receptor (RyR1). Certain RyR1 mutations result in an MH-selective phenotype (MH-only), whereas others result in a mixed phenotype (MH + CCD). We characterized effects on Ca2+ handling and excitation-contraction (EC) coupling of MH-only and MH + CCD mutations in RyR1 after expression in skeletal myotubes derived from RyR1-null (dyspedic) mice. Compared to wild-type RyR1-expressing myotubes, MH + CCD- and MH-only-expressing myotubes exhibited voltage-gated Ca2+ release (VGCR) that activated at more negative potentials and displayed a significantly higher incidence of spontaneous Ca2+ oscillations. However, maximal VGCR was reduced only for MH + CCD mutants (Y4795C, R2435L, and R2163H) in which spontaneous Ca2+ oscillations occurred with significantly longer duration (Y4795C and R2435L) or higher frequency (R2163H). Notably, myotubes expressing these MH + CCD mutations in RyR1 exhibited both increased [Ca2+]i and reduced sarcoplasmic reticulum (SR) Ca2+ content. We conclude that MH-only mutations modestly increase basal release-channel activity in a manner insufficient to alter net SR Ca2+ content ("compensated leak"), whereas the mixed MH + CCD phenotype arises from mutations that enhance basal activity to a level sufficient to promote SR Ca2+ depletion, elevate [Ca2+]i, and reduce maximal VGCR ("decompensated leak"). PMID- 15347587 TI - Cytochrome c adsorption to supported, anionic lipid bilayers studied via atomic force microscopy. AB - The adsorption of membrane-associated protein cytochrome c to anionic lipid bilayers of dioleoyl phosphatidylglycerol was studied in low ionic strength physiological buffer using atomic force microscopy. The bilayers were supported on polylysinated mica. The formation of stable, single lipid bilayers was confirmed by imaging and force spectroscopy. Upon addition of low concentrations of cytochrome c, protein molecules were not topographically visible on the lipid bilayer-buffer interface. However, the forces required to punch through the bilayer by indentation using the atomic force microscopy probe were significantly lower after protein adsorption, which suggest that the protein inserts into the bilayer. Moreover, the apparent thickness of the bilayer remained unchanged after cytochrome c adsorption. Yet, mass spectroscopy and visible light absorption spectroscopy confirmed the presence of cytochrome c in the lipid bilayers. These results suggest that 1), cytochrome c inserts into the bilayer and resides in its hydrophobic core; 2), cytochrome c insertion changes the mechanical properties of the bilayer significantly; and 3), bilayer force spectroscopy may be a useful tool in investigating lipid-protein interactions. PMID- 15347588 TI - Role of the linker domain and the 203-214 N-terminal residues in the human topoisomerase I DNA complex dynamics. AB - The influence of the N-terminal residues 203-214 and the linker domain on motions in the human topoisomerase I-DNA complex has been investigated by comparing the molecular dynamics simulations of the system with (topo70) or without (topo58/6.3) these regions. Topo58/6.3 is found to fluctuate more than topo70, indicating that the presence of the N-terminal residues and the linker domain dampen the core and C-terminal fluctuations. The simulations also show that residues 203-207 and the linker domain participate in a network of correlated movements with key regions of the enzyme, involved in the human topoisomerase I catalytic cycle, providing a structural-dynamical explanation for the better DNA relaxation activity of topo70 when compared to topo58/6.3. The data have been examined in relation to a wealth of biochemical, site-directed mutagenesis and crystallographic data on human topoisomerase I. The simulations finally show the occurrence of a network of direct and water mediated hydrogen bonds in the proximity of the active site, and the presence of a water molecule in the appropriate position to accept a proton from the catalytic Tyr-723 residue, suggesting that water molecules have an important role in the stabilization and function of this enzyme. PMID- 15347589 TI - Binding site in eag voltage sensor accommodates a variety of ions and is accessible in closed channel. AB - In ether-a-go-go K+ channels, voltage-dependent activation is modulated by ion binding to a site located in an extracellular-facing crevice between transmembrane segments S2 and S3 in the voltage sensor. We find that acidic residues D278 in S2 and D327 in S3 are able to coordinate a variety of divalent cations, including Mg2+, Mn2+, and Ni2+, which have qualitatively similar functional effects, but different half-maximal effective concentrations. Our data indicate that ions binding to individual voltage sensors in the tetrameric channel act without cooperativity to modulate activation gating. We have taken advantage of the unique phenotype of Ni2+ in the D274A channel, which contains a mutation of a nonbinding site residue, to demonstrate that ions can access the binding site from the extracellular solution when the voltage sensor is in the resting conformation. Our results are difficult to reconcile with the x-ray structure of the KvAP K+ channel, in which the binding site residues are widely separated, and with the hydrophobic paddle model for voltage-dependent activation, in which the voltage sensor domain, including the S3-S4 loop, is near the cytoplasmic side of the membrane in the closed channel. PMID- 15347590 TI - Modeling feedback loops of the Mammalian circadian oscillator. AB - The suprachiasmatic nucleus governs daily variations of physiology and behavior in mammals. Within single neurons, interlocked transcriptional/translational feedback loops generate circadian rhythms on the molecular level. We present a mathematical model that reflects the essential features of the mammalian circadian oscillator to characterize the differential roles of negative and positive feedback loops. The oscillations that are obtained have a 24-h period and are robust toward parameter variations even when the positive feedback is replaced by a constantly expressed activator. This demonstrates the crucial role of the negative feedback for rhythm generation. Moreover, it explains the rhythmic phenotype of Rev-erbalpha-/- mutant mice, where a positive feedback is missing. The interplay of negative and positive feedback reveals a complex dynamics. In particular, the model explains the unexpected rescue of circadian oscillations in Per2Brdm1/Cry2-/- double-mutant mice (Per2Brdm1 single-mutant mice are arrhythmic). Here, a decrease of positive feedback strength associated with mutating the Per2 gene is compensated by the Cry2-/- mutation that simultaneously decreases the negative feedback strength. Finally, this model leads us to a testable prediction of a molecular and behavioral phenotype: circadian oscillations should be rescued when arrhythmic Per2Brdm1 mutant mice are crossed with Rev- erbalpha -/- mutant mice. PMID- 15347591 TI - Cholesterol depletion increases membrane stiffness of aortic endothelial cells. AB - This study has investigated the effect of cellular cholesterol on membrane deformability of bovine aortic endothelial cells. Cellular cholesterol content was depleted by exposing the cells to methyl-beta-cyclodextrin or enriched by exposing the cells to methyl-beta-cyclodextrin saturated with cholesterol. Control cells were treated with methyl-beta-cyclodextrin-cholesterol at a molar ratio that had no effect on the level of cellular cholesterol. Mechanical properties of the cells with different cholesterol contents were compared by measuring the degree of membrane deformation in response to a step in negative pressure applied to the membrane by a micropipette. The experiments were performed on substrate-attached cells that maintained normal morphology. The data were analyzed using a standard linear elastic half-space model to calculate Young elastic modulus. Our observations show that, in contrast to the known effect of cholesterol on membrane stiffness of lipid bilayers, cholesterol depletion of bovine aortic endothelial cells resulted in a significant decrease in membrane deformability and a corresponding increase in the value of the elastic coefficient of the membrane, indicating that cholesterol-depleted cells are stiffer than control cells. Repleting the cells with cholesterol reversed the effect. An increase in cellular cholesterol to a level higher than that of normal cells, however, had no effect on the elastic properties of bovine aortic endothelial cells. We also show that although cholesterol depletion had no apparent effect on the intensity of F-actin-specific fluorescence, disrupting F actin with latrunculin A abrogated the stiffening effect. We suggest that cholesterol depletion increases the stiffness of the membrane by altering the properties of the submembrane F-actin and/or its attachment to the membrane. PMID- 15347592 TI - Mobile actin clusters and traveling waves in cells recovering from actin depolymerization. AB - At the leading edge of a motile cell, actin polymerizes in close apposition to the plasma membrane. Here we ask how the machinery for force generation at a leading edge is established de novo after the global depolymerization of actin. The depolymerization is accomplished by latrunculin A, and the reorganization of actin upon removal of the drug is visualized in Dictyostelium cells by total internal reflection fluorescence microscopy. The actin filament system is reorganized in three steps. First, F-actin assembles into globular complexes that move along the bottom surface of the cells at velocities up to 10 microm/min. These clusters are transient structures that eventually disassemble, fuse, or divide. In a second step, clusters merge into a contiguous zone at the cell border that spreads and gives rise to actin waves traveling on a planar membrane. Finally, normal cell shape and motility are resumed. These data show that the initiation of actin polymerization is separated in Dictyostelium from front protrusion, and that the coupling of polymerization to protrusion is a later step in the reconstitution of a leading edge. PMID- 15347593 TI - Nanopore unzipping of individual DNA hairpin molecules. AB - We have used the nanometer scale alpha-Hemolysin pore to study the unzipping kinetics of individual DNA hairpins under constant force or constant loading rate. Using a dynamic voltage control method, the entry rate of polynucleotides into the pore and the voltage pattern applied to induce hairpin unzipping are independently set. Thus, hundreds of unzipping events can be tested in a short period of time (few minutes), independently of the unzipping voltage amplitude. Because our method does not entail the physical coupling of the molecules under test to a force transducer, very high throughput can be achieved. We used our method to study DNA unzipping kinetics at small forces, which have not been accessed before. We find that in this regime the static unzipping times decrease exponentially with voltage with a characteristic slope that is independent of the duplex region sequence, and that the intercept depends strongly on the duplex region energy. We also present the first nanopore dynamic force measurements (time varying force). Our results are in agreement with the approximately logV dependence at high V (where V is the loading rate) observed by other methods. The extension of these measurements to lower loading rates reveals a much weaker dependence on V. PMID- 15347594 TI - Coupling field theory with mesoscopic dynamical simulations of multicomponent lipid bilayers. AB - A method for simulating a two-component lipid bilayer membrane in the mesoscopic regime is presented. The membrane is modeled as an elastic network of bonded points; the spring constants of these bonds are parameterized by the microscopic bulk modulus estimated from earlier atomistic nonequilibrium molecular dynamics simulations for several bilayer mixtures of DMPC and cholesterol. The modulus depends on the composition of a point in the elastic membrane model. The dynamics of the composition field is governed by the Cahn-Hilliard equation where a free energy functional models the coupling between the composition and curvature fields. The strength of the bonds in the elastic network are then modulated noting local changes in the composition and using a fit to the nonequilibrium molecular dynamics simulation data. Estimates for the magnitude and sign of the coupling parameter in the free energy model are made treating the bending modulus as a function of composition. A procedure for assigning the remaining parameters in the free energy model is also outlined. It is found that the square of the mean curvature averaged over the entire simulation box is enhanced if the strength of the bonds in the elastic network are modulated in response to local changes in the composition field. We suggest that this simulation method could also be used to determine if phase coexistence affects the stress response of the membrane to uniform dilations in area. This response, measured in the mesoscopic regime, is already known to be conditioned or renormalized by thermal undulations. PMID- 15347595 TI - Molecular basis for the dynamic strength of the integrin alpha4beta1/VCAM-1 interaction. AB - Intercellular adhesion mediated by integrin alpha4beta1 and vascular cell adhesion molecule-1 (VCAM-1) plays a crucial role in both the rolling and firm attachment of leukocytes onto the vascular endothelium. Essential to the alpha4beta1/VCAM-1 interaction is its mechanical strength that allows the complex to resist the large shear forces imposed by the bloodstream. Herein we employed single-molecule dynamic force spectroscopy to investigate the dynamic strength of the alpha4beta1/VCAM-1 complex. Our force measurements revealed that the dissociation of the alpha4beta1/VCAM-1 complex involves overcoming at least two activation potential barriers: a steep inner barrier and a more elevated outer barrier. The inner barrier grants the complex the tensile strength to withstand large pulling forces (>50 pN) and was attributed to the ionic interaction between the chelated Mg2+ ion at the N-terminal A-domain of the beta1 subunit of alpha4beta1 and the carboxyl group of Asp-40 of VCAM-1 through the use of site directed mutations. In general, additional mutations within the C-D loop of domain 1 of VCAM-1 suppressed both inner and outer barriers of the alpha4beta1/VCAM-1 complex, while a mutation at Asp-143 of domain 2 of VCAM-1 resulted in the suppression of the outer barrier, but not the inner barrier. In contrast, the outer barrier of alpha4beta1/VCAM-1 complex was stabilized by integrin activation. Together, these findings provide a molecular explanation for the functionally relevant kinetic properties of the alpha4beta1/VCAM-1 interaction. PMID- 15347596 TI - First polar body morphology before ICSI is not related to embryo quality or pregnancy rate. AB - BACKGROUND: The aim of this study was to analyse the relationship between the first polar body (1st PB) morphology and the fertilization rate, cleavage rate, embryo quality, pregnancy and implantation rate. METHODS: This was a retrospective study on 167 consecutive cycles undergoing assisted reproduction with ICSI. The 1st PB morphology was evaluated at the moment of ICSI in the 596 injected oocytes and it was coded as intact or fragmented. The fertilization rate, cleavage rate, embryo quality (three grades), pregnancy rate, implantation rate and the time elapsed between oocyte retrieval and ICSI were evaluated. The 1st PB morphology was checked twice (denudation and ICSI) in a random sample of 180 oocytes in order to verify the effect of the in vitro culture. RESULTS: No significant relationship was found between the 1st PB morphology and the fertilization rate (P=0.703), cleavage rate (P=0.055), embryo quality (P=0.673), pregnancy rate (P=0.201) and implantation rate (P=0.511). A significant positive relationship (P=0.006) was found between the frequency of the 1st PB fragmentation and the time elapsed between denudation and ICSI. The pregnancy rate was significantly higher (P=0.008) when oocytes were injected between 5 and 7 h after retrieval rather than earlier or later. CONCLUSIONS: Our data suggest that the embryo quality, pregnancy rate and implantation rate are not related to the 1st PB fragmentation. The time which elapses between the oocyte retrieval and ICSI should be maintained at approximately 6 h in order to obtain optimal results. PMID- 15347597 TI - Blastocoele collapse by micropipetting prior to vitrification gives excellent survival and pregnancy outcomes for human day 5 and 6 expanded blastocysts. AB - BACKGROUND: Manual puncture of the trophectoderm of human blastocysts with a needle before vitrification increases their survival rate, but the embryos take a long time to re-expand. This study examined whether causing human blastocysts to collapse by manual pipetting before vitrification would allow more rapid re expansion and improve pregnancy rates. METHODS: After embryo transfer in IVF cycles, surplus embryos that developed to the expanded blastocyst stage were placed in cryoprotectant and then artificially shrunk by mechanical pipetting with a fine hand-drawn glass pipette slightly smaller in diameter than the blastocyst. The shrunken embryos were placed in a small volume of vitrification solution and plunged into liquid nitrogen on a cryotop. The blastocysts were thawed by warming and then dilution in 1 mol/l sucrose. RESULTS: Of 49 expanded vitrified blastocysts, 48 (98%) re-expanded within 3 h after warming. Following transfer (48 blastocysts in 28 cycles), 14 women (50%) became clinically pregnant, and the implantation rate was 33% (16/48). Eight healthy babies have been born in six deliveries, and the other eight pregnancies are ongoing. To date, there have been no spontaneous abortions. CONCLUSIONS: The results suggest that artificial shrinkage with pipetting is a simple and effective technique to assist successful cryopreservation of expanded blastocysts by vitrification. PMID- 15347598 TI - Mouse oocyte meiotic resumption and polar body extrusion in vitro are differentially influenced by FSH, epidermal growth factor and meiosis-activating sterol. AB - BACKGROUND: In this study, we compared the relative ability of FSH (100 mIU/ml), epidermal growth factor (EGF) (10 ng/ml), and follicular-fluid meiosis-activating sterol (FF-MAS, 10 micromol/l) to induce meiotic resumption and polar body I (PBI) extrusion in mouse oocytes. METHODS: Cumulus-enclosed oocytes (CEO) were co incubated with meiosis-arresting agents, including 4 mmol/l hypoxanthine (Hx), 0.3 mmol/l dibutyryl cAMP (dbcAMP), and 8.5 micromol/l cilostamide, a selective inhibitor of the oocyte-specific phosphodiesterase 3 (PDE 3). RESULTS: In Hx treated oocytes, FSH, EGF and FF-MAS induced meiosis resumption at very high rates, but only FSH and EGF also promoted PBI extrusion with high frequency. In experiments conducted in the presence of dbcAMP, FF-MAS was unable to promote an increase in germinal vesicle breakdown (GVBD) rate, whereas FSH and EGF generated a response similar to the Hx groups. Neither FSH, EGF nor FF-MAS caused any change in the meiotic status of CEO when meiotic arrest at the germinal vesicle (GV) stage was maintained by cilostamide. In the presence of Hx, naked oocytes (NkO) co-cultured with their cumulus cells were able to respond to the GVBD inducing effect of FSH and EGF by resuming meiosis at high rate. CONCLUSIONS: Collectively, these results indicate that: (i) a signal triggered in cumulus cells by either FSH or EGF, but not necessarily coincident with FF-MAS, may contribute to meiotic maturation, supporting GVBD and extrusion of PBI; (ii) the transmission of this signal can occur in a paracrine fashion, at least with reference to the breakdown of the GV. It also appears that concomitant regulation of intra-oocyte cAMP degradation is a prerequisite for meiosis resumption. PMID- 15347599 TI - Organ-selective chemoresistance in metastasis from human breast cancer cells: inhibition of apoptosis, genetic variability and microenvironment at the metastatic focus. AB - We hypothesized that the development of the most resistant cells during metastasis is favored by anti-apoptotic proteins, leading to the acquisition of an adaptive phenotype crucial to drug resistance at the metastatic foci. In order to test it, we induced metastasis in nude mice, injecting orthotopicaly 435/Bcl x(L) or 435/Neo cells, transfected previously with the luciferase gene to use it as a tumor marker, and treated them with a therapeutic dose of docetaxel. We monitored metastasis in mice by calculating tumor cell equivalents (TCEs) present in tissues. Between docetaxel-treated and non-treated 435/Bcl-x(L).luc mice significant differences in the metastatic burden of lymph nodes (P = 0.02) and viscera (P = 0.02) were observed. However, treatment did not significantly decrease metastatic burden in bones (P = 0.19). Additionally, we analyzed the clonality of metastasis from lung, bone and lymph node by genomic DNA fingerprinting. Bcl-x(L) enhanced cell genetic instability in terms of gain and loss fractions (GF = 0.18 and LF = -0.21) when compared with the control 435/Neo (GF = 0.15 and LF = -0.14). Thus, genetic instability might be a molecular mechanism favored by Bcl-x(L) evolved in the selection process of breast cancer progression, which results in different genetic changes among metastases from lung, bone or lymph node, favoring organ-selective chemoresistance. PMID- 15347600 TI - Cell cycle kinase inhibitor expression and hypoxia-induced cell cycle arrest in human cancer cell lines. AB - Flow cytometric analysis of fibroblasts, normal breast epithelial cells and breast or other cancer cell lines identified variation in the abilities of cell lines to undergo cell cycle arrest as a response to hypoxia. Human mammary epithelial cells (HMEC), normal fibroblasts (Hs68 and WI38), HeLa cervical carcinoma and HTB-30 breast carcinoma cells arrest in G(1)/S in response to severe hypoxia. Hep3B hepatocellular carcinoma cells did not exhibit orderly G(1)/S arrest in response to severe hypoxia. We found a general decrease in p16(INK4a) (p16) mRNA levels, with an associated decrease in p16 protein levels in both normal cells and in cancer cells, regardless of their cell cycle response to hypoxia. p27 protein levels did not correlate with the cell line's ability to enter a hypoxic G(1)/S arrest. Furthermore, cell lines that underwent G(1)/S arrest showed decreased expression of hypoxia inducible factor 1 (HIF-1alpha) and at least one member of INK4 or Sdi cell cycle kinase inhibitors families after 12 24 h of hypoxia. Conversely, Hep3B, which did not exhibit orderly hypoxia associated G(1)/S arrest, also did not show decreased HIF-1alpha, INK4 or Sdi protein levels in hypoxia. Furthermore, Hep3B showed constitutive activating phosphorylation of Akt and inhibitory phosphorylation of GSK3beta, which was the opposite pattern to that exhibited by the cell lines showing the G(1)/S arrest phenotype. Inhibition of GSK3beta by lithium chloride treatment of HeLa cells converted the HIF-1alpha, p16 and p27 loss to levels unchanged by hypoxic exposure. Our results suggest that regulation of the cell cycle during hypoxia in either normal or cancer cells is not simply due to up-regulation of cell cycle kinase inhibitors. Furthermore, decreased protein expression of HIF-1alpha, p16 and p27 was associated with both a hypoxia-induced G(1)/S arrest phenotype and increased GSK3beta activity. PMID- 15347601 TI - Mutations in TP53, but not FGFR3, in urothelial cell carcinoma of the bladder are influenced by smoking: contribution of exogenous versus endogenous carcinogens. AB - Smoking is a major risk factor for urothelial cell carcinoma of the bladder (UCC). Mutations in the FGFR3 and TP53 genes have been shown to define two distinct pathways in superficial papillary and invasive UCC disease, respectively. We investigated the relationship between smoking and these mutations by means of denaturing high performance liquid chromatography and sequencing for 110 primary UCC of the bladder. This study included 48 current smokers, 31 ex-smokers and 31 non-smokers. Thirty-five of the tumors were stage pTa, 40 pT1 and 35 > or =pT2. Fourteen of the tumors were grade 1, 37 were grade 2 and 59 grade 3. Smoking was associated with high stage (P = 0.03) and high grade tumors (P = 0.006). Twenty-two of the 110 tumors studied harbored TP53 mutations (20%) and 43 harbored FGFR3 mutations (39%). Odds ratios (OR) were higher for TP53 mutations in current smokers [OR, 2.25; 95% confidence interval (95% CI), 0.65-7.75] and ex-smokers (OR, 1.62; 95% CI, 0.41-6.42) than in non smokers. Double TP53 mutations and the A:T-->G:C TP53 mutation pattern was found only in current smokers. Patients with the FGFR3(wild-type)/TP53(mutated) genotype had significantly higher levels of tobacco consumption, as measured in pack-years (P = 0.01). Smoking influenced neither the frequency nor the pattern of FGFR3 mutations. Our results suggest that smoking is associated with invasive and high grade UCCs, at initial presentation, and influenced TP53 or the molecular pathway defined by these mutations. In contrast, FGFR3 mutations are not affected by smoking and probably result from endogenous alterations. These data have potential implications for clinical management and prevention strategies. PMID- 15347602 TI - Reduction in mortality from severe head injury following introduction of a protocol for intensive care management. AB - BACKGROUND: To determine the effect of an intensive care management protocol on the intensive care unit (ICU) and hospital mortality of severely head-injured patients, we designed a longitudinal observational study of all patients admitted with a head injury between 1992 and 2000. METHODS: A computerized patient database was used to identify all patients with severe head injury admitted to the ICU at Frenchay Hospital, Bristol, UK: a tertiary referral centre for the clinical neurosciences. We compared the ICU and hospital mortality and length of stay in patients before and after implementation of a protocol for their ICU management in 1997. RESULTS: Implementation of the protocol was associated with a significant reduction in ICU mortality from 19.95% to 13.5% (odds ratio 0.47; 95% CI 0.29-0.75), and in hospital mortality from 24.55% to 20.8% (odds ratio 0.48; 95% CI 0.31-0.74). This was achieved despite a significant increase in the median APACHE II score (14 vs 18) of patients admitted after implementation of the protocol. The median ICU and hospital length of stay remained constant over the study period. CONCLUSIONS: The introduction of an evidence-based protocol to guide the ICU management of patients with severe head injury has been associated with a significant reduction in both ICU and hospital mortality. PMID- 15347603 TI - Chronic ethanol consumption does not affect action of propofol on rat hippocampal acetylcholine release in vivo. AB - BACKGROUND: The aim of this study was to examine ethanol-consumption-related changes in the effects of propofol on rat hippocampal acetylcholine (ACh) release. METHODS: Male Sprague-Dawley rats received a solution of ethanol (20% v/v) for 24 weeks while controls received tap water. The effects of propofol were examined by in vivo microdialysis, with ACh release from the hippocampal regions determined by high-performance liquid chromatography with electrochemical detection (HPLC-ECD). RESULTS: Propofol 50 mg kg(-1) i.p. significantly decreased basal hippocampal ACh release in ethanol-treated and control rats by 50.4 (sem 4.7)% and 38.3 (11.1)%, respectively. Propofol 100 mg kg(-1) i.p. significantly decreased basal hippocampal ACh release in ethanol-treated and control rats by 67.5 (3.7)% and 55.9 (7.4)%, respectively. The reduction in hippocampal ACh release induced by 50 or 100 mg kg(-1) i.p. propofol was not significantly different between ethanol-treated and control rats. There was no significant difference in the duration of sleep between the two groups. CONCLUSIONS: These results demonstrate that chronic ethanol consumption does not augment the inhibitory actions of propofol on rat hippocampal ACh release. These findings appear to be inconsistent with the notion that chronic ethanol intake enhances the propofol-induced inhibition of the hippocampal cholinergic system and related mental dysfunction. PMID- 15347604 TI - High concentrations of N-BNP are related to non-infectious severe SIRS associated with cardiovascular dysfunction occurring after off-pump coronary artery surgery. AB - BACKGROUND: Procalcitonin (PCT) blood concentrations are known to be an appropriate marker of severe systemic inflammatory response syndrome (SIRS) induced by coronary artery surgery with and without cardiopulmonary bypass. Pro brain natriuretic peptide (N-BNP) is a newly described cardiac hormone considered to be an effective marker of severity and prognosis of acute coronary syndromes and congestive heart failure. We evaluated the perioperative time courses of PCT and N-BNP and investigated their role as early markers of severe SIRS (SIRS with cardiovascular dysfunction) induced by off-pump coronary artery bypass (OPCAB). METHODS: Sixty-three patients were prospectively included. The American College of Chest Physicians Classification was used to diagnose SIRS and organ system failure to define severe SIRS. Serum concentrations of PCT and N-BNP were determined before, during and after surgery. Receiver operating characteristic curves and cut-off values were used to assess the ability of these markers to predict postoperative severe SIRS. RESULTS: SIRS occurred in 25 (39%) patients. Nine of them (14%) showed severe SIRS. Significantly higher serum concentrations of N-BNP and PCT were found in patients with severe SIRS with peak concentrations respectively at 8887 pg ml(-1) (range 2940-29372 pg ml(-1)) for N-BNP and 9.50 ng ml(-1) (range 1-65 ng ml(-1)) for PCT. The area under the curve using N-BNP to detect postoperative severe SIRS was 0.799 before surgery (0.408 for PCT; P<0.01) and 0.824 at the end of surgery (0.762 for PCT; P<0.05). CONCLUSIONS: N-BNP may be an appropriate marker indicating the early development of non-infectious postoperative severe SIRS after OPCAB. PMID- 15347605 TI - Effects of milrinone on jugular bulb oxygen saturation and cerebrovascular carbon dioxide reactivity in patients undergoing coronary artery bypass graft surgery. AB - BACKGROUND: Jugular bulb oxygen saturation (Sjv(o(2))) is a surrogate marker for global cerebral oxygenation. The effect of milrinone on Sjv(o(2)) and the cerebrovascular carbon dioxide reactivity (CCO2R) was investigated. METHODS: Thirty patients scheduled for coronary artery bypass graft surgery (CABG) were studied prospectively. After sternotomy, normoventilation (at T(1); Pa(co(2))=4.7 5.0 kPa) and hyperventilation (at T(2); Pa(co(2))=3.3-3.7 kPa) were induced and the changes in Sjv(o(2)) (DeltaSjv(o(2))) and Pa(co(2)) (DeltaPa(co(2))), and DeltaSjv(o(2))/DeltaPa(co(2)) (CCO(2)R) were measured. After normoventilation was re-established (at T(3)), milrinone 50 microg kg(-1) was given (at T(4)), followed by hyperventilation (at T(5)), and DeltaSjv(o(2)), DeltaPa(co(2)) and CCO(2)R were measured. RESULTS: After milrinone administration at normoventilation (T(3) and T(4)), cardiac index and mixed venous oxygen saturation increased, while mean arterial pressure and systemic vascular resistance index decreased, without a significant change in Sjv(o(2)). Before milrinone administration (T(1) and T(2)), hyperventilation decreased Pa(co(2)) and Sjv(o(2)), and DeltaSjv(o(2)) showed positive linear correlation with DeltaPa(co(2)). After milrinone administration (T(4) and T(5)), hyperventilation decreased Pa(co(2)) and Sjv(o(2)), and DeltaSjv(o(2)) showed positive linear correlation with DeltaPa(co(2)). There was no significant difference in CCO(2)R before and after milrinone administration (13.3 (5.7)% kPa(-1) and 12.3 (3.9)% kPa(-1), respectively). CONCLUSIONS: Although milrinone induced significant haemodynamic changes, Sjv(o(2)) and CCO(2)R were unchanged during its administration. PMID- 15347606 TI - Recent advances in intravenous anaesthesia. AB - Efforts to develop new hypnotic compounds continue, although several have recently failed in development. Propofol has been reformulated in various presentations with and without preservatives. Pharmacokinetic and pharmacodynamic differences exist between some of these preparations, and it is currently unclear whether any have substantial advantages over the original presentation. The use of target-controlled infusion (TCI) has been extended to include paediatric anaesthesia and sedation. Application of TCI to remifentanil is now licensed. Linking of electroencephalogram (EEG) monitoring to TCI for closed-loop anaesthesia remains a research tool, although commercial development may follow. The availability of stereoisomer ketamine and improved understanding of its pharmacology have increased non-anaesthetic use of ketamine as an adjunct analgesic. It may be useful in subhypnotic doses for postsurgical patients with pain refractory to morphine administration. PMID- 15347607 TI - Anaesthetic management of the newborn with multiple congenital epulides. AB - Epulis of the newborn is a granular cell tumour that originates from the dental alveolar mucosa. We report a case of a neonate with multiple congenital masses of the alveolar mucosa who presented for surgery with a potential airway problem. Intubation was achieved uneventfully using a gaseous induction with a large facemask and displacement of the epulides to allow cautious laryngoscopy. PMID- 15347608 TI - Comparison of hyperbaric and plain ropivacaine 15 mg in spinal anaesthesia for lower limb surgery. AB - BACKGROUND: Previously, plain ropivacaine 15 mg given intrathecally has been shown to be feasible for ambulatory surgery of lower-extremities. Hypothetically, hyperbaric solution could improve and shorten the block. METHODS: This prospective, randomized, double-blind study included 56 patients undergoing surgery of lower extremities. They received intrathecally either 1.5 ml of ropivacaine 10 mg ml(-1) and 0.5 ml of glucose 300 mg ml(-1) (HYP) or 2 ml of ropivacaine 7.5 mg ml(-1) (PL). RESULTS: All patients in Group HYP achieved T(10) dermatome analgesia but only 64% (18/28) of Group PL. T(10) analgesia was reached in 5 min (median, range 5-20 min) in the HYP group vs 10 min (5-45 min) in the PL group (P=0.022), and full motor block in 10 min (5-45 min) vs 20 min (5-60 min) (P=0.003), respectively. Group HYP had a longer duration of analgesia at T(10); 83 min (5-145 min) vs 33 min (0-140 min) (P=0.004). Duration of sensory block from injection of the anesthetic to complete recovery was shorter in Group HYP than in Group PL, 210 min (120-270 min) vs 270 min (210-360 min) (P<0.001), as was duration of motor block, 120 min (5-150 min) vs 210 min (120-330 min) (P<0.001). Patients of Group HYP attained discharge criteria earlier than those of Group PL (P=0.009). CONCLUSION: In comparison with the plain solution, 15 mg of intrathecal hyperbaric ropivacaine produced a faster onset, greater success rate of analgesia at the level of T(10) dermatome, and faster recovery of the block. PMID- 15347609 TI - Spread of subarachnoid block, intraoperative local anaesthetic requirements and postoperative analgesic requirements in Caesarean section and total abdominal hysterectomy. AB - BACKGROUND: Pregnancy is associated with a higher spread of subarachnoid anaesthesia and increased pain threshold. The study was designed to assess the spread of subarachnoid block and the intra- and postoperative analgesic requirements in pregnant vs non-pregnant women. METHODS: We assessed the level of subarachnoid anaesthesia after 1.8 ml of hyperbaric lidocaine 5% and the postoperative analgesic requirements in women undergoing Caesarean section and undergoing abdominal hysterectomy (30 each group). Intraoperatively epidural ropivacaine was given as required. All patients received 10 ml of ropivacaine 0.2% epidurally 2, 10, and 24 h after operation and the VAS pain score was assessed. They also had access to patient controlled analgesia i.v. morphine. RESULTS: Duration of surgery was 64 (13.7) vs 127 (33.8) min (P<0.0001) in the pregnant and non-pregnant groups. Ten minutes after subarachnoid injection, sensory block was higher by three dermatomes in the pregnant group (P<0.0001). Time to first ropivacaine dose was 37 (19.7) vs 19 (12.2) min (P<0.001) and the ropivacaine normalized for the duration of anaesthesia was 0.8 (0.6) vs 1.3 (0.5) mg(-1) (P=0.001) in the pregnant and non-pregnant groups, respectively. The time between the first and second ropivacaine dose was similar in the two groups (P=0.070). Fewer pregnant women (81 vs 100%) required ropivacaine intraoperatively (P=0.017). The VAS scores were similar but parturients consumed more i.v. morphine (33 (14) vs 24 (12) mg, P=0.016) during the first 24 h after operation. CONCLUSIONS: Pregnant patients exhibited a higher level of subarachnoid sensory block and required more i.v. morphine after operation. PMID- 15347610 TI - Effect of lidocaine on ischaemic preconditioning in isolated rat heart. AB - BACKGROUND: Lidocaine is frequently used as an agent to treat ventricular arrhythmias associated with acute myocardial ischaemia. Lidocaine is a potent blocker not only of sodium channels, but also of ATP-sensitive potassium channels. The opening of these channels is a key mechanism of ischaemic preconditioning. We investigated the hypothesis that lidocaine blocks the cardioprotection induced by ischaemic preconditioning. METHODS: Isolated rat hearts (n=60) were subjected to 30 min of no-flow ischaemia and 60 min of reperfusion. Control hearts (CON) underwent no further intervention. Preconditioned hearts (PC) received two 5-min periods of ischaemia separated by 10 min of reflow before the 30 min ischaemia. In three groups, lidocaine was infused at concentrations of 2, 10 or 20 microg ml(-1) for 5 min before the preconditioning ischaemia. Left ventricular developed pressure (LVDP) and infarct size (IS) (triphenyltetrazolium choride staining) were measured as variables of ventricular function and cellular injury, respectively. RESULTS: PC reduced IS from 24.8 (sem 4.1) % to 4.0 (0.7) % of the area at risk (P<0.05). Adding 2 or 10 microg ml(-1) lidocaine had no effect on IS compared with PC alone (3.7 (0.7) %, 6.9 (1.8) %). Adding 20 microg ml(-1) lidocaine increased IS to 14.1 (2.5) % compared with PC (P<0.05). Baseline LVDP was similar in all groups (111.4 (2.1) mm Hg). Compared with CON, PC improved functional recovery (after 60 min of reperfusion; 52.3 (5.9) mm Hg vs 16.0 (4.0) mm Hg, P<0.01). The improved ventricular function was not influenced by addition of 2 or 10 microg ml(-1) lidocaine (47.3 (5.7) mm Hg, not significant; 45.3 (7.3) mm Hg, not significant), but was blocked by the infusion of 20 microg ml(-1) lidocaine (22.5 (8.0) mm Hg, P<0.01 vs PC). CONCLUSIONS: Lidocaine blocks the cardioprotection induced by ischaemic preconditioning only at supratherapeutic concentrations. PMID- 15347611 TI - Comment on the "Report of the AAHA Canine Vaccine Task Force: 2003 canine vaccine guidelines, recommendations, and supporting literature". PMID- 15347613 TI - Obtaining quality biopsies of the liver and kidney. PMID- 15347614 TI - Spectrum of M-mode echocardiographic abnormalities in 75 cats with systemic hypertension. AB - A retrospective study was performed in 75 hypertensive cats to determine the spectrum and frequency of M-mode echocardiographic abnormalities. Results indicated that 21.3% of the cats had M-mode measurements within normal reference ranges. For cats with echocardiographic abnormalities, changes were variable. Thirty-nine percent of hypertensive cats had hypertrophy of the interventricular septum in diastole, and 41.3% had hypertrophy of the left ventricular (LV) posterior wall in diastole. One cat in five had a dilated left atrium, while fractional shortening and LV internal dimension in diastole were normal in 82.7% and 86.7% of the cats, respectively. The marked variability of echocardiographic findings in hypertensive cats made echocardiography an unreliable screening test for hypertension. PMID- 15347615 TI - Lactate as a diagnostic test for septic peritoneal effusions in dogs and cats. AB - Lactate concentration in peritoneal fluid was evaluated and compared to blood lactate concentration in dogs and cats with septic and nonseptic abdominal effusions. All dogs with septic effusions had a peritoneal fluid lactate concentration >2.5 mmol/L and a peritoneal fluid lactate concentration higher than blood lactate, resulting in a negative blood to fluid lactate difference. In dogs, the diagnostic accuracy of the peritoneal fluid lactate concentration and the blood to fluid lactate difference in differentiating septic peritoneal effusion was 95% and 90%, respectively. Peritoneal fluid lactate concentration and blood to fluid lactate difference were not accurate tests for detecting septic peritoneal effusions in cats. PMID- 15347616 TI - Biochemical analysis of neoplastic versus nonneoplastic abdominal effusions in dogs. AB - This prospective study of 15 dogs evaluated biochemical parameters in abdominal effusions of neoplastic and nonneoplastic origin in an attempt to identify markers for malignant effusions. Dogs in the neoplastic group had statistically significant lower glucose concentrations (mean, 72.6 mg/dL versus 110.0 mg/dL; P=0.0431) and higher lactate levels (mean, 3.81 mmol/L versus 1.68 mmol/L; P=0.0377) in their abdominal fluid than did dogs in the nonneoplastic group, indicating that low glucose and high lactate in abdominal effusions may be markers for neoplasia. PMID- 15347617 TI - Comparison of serological tests for the detection of natural heartworm infection in cats. AB - Serological tests were performed on 380 cats with necropsy-confirmed heartworm disease to compare the performance of currently available commercial laboratory and point-of-care heart-worm serological tests in a heartworm-endemic area. Overall, antigen tests detected 79.3% to 86.2% of heartworm infections and were highly specific. Most cats with false-negative antigen tests had a single male worm. Antibody tests detected 62.1% to 72.4% of heartworm infections and had a wider range of false-positive results (1.4% to 19.1%) than antigen tests (0.3% to 2.0%). Serological tests for feline heartworm infection varied in diagnostic performance. Combining results from antigen and antibody tests achieved greater sensitivity than using either test alone. PMID- 15347618 TI - Cranial cruciate ligament pathophysiology in dogs with cruciate disease: a review. AB - Cruciate disease is a common cause of chronic lameness in dogs. Midsubstance rupture of the cranial cruciate ligament (CCL) arises from progressive pathological failure, often under conditions of normal loading in adult dogs with CCL instability. A high risk of rupture is associated with inflammation of the synovium and adaptive or degenerative changes in the cells and matrix of the CCL. In contrast, CCL rupture in puppies is usually associated with traumatic injury and avulsion of the CCL from its sites of attachment. PMID- 15347619 TI - C-reactive protein in the differentiation of pyometra from cystic endometrial hyperplasia/mucometra in dogs. AB - Hematological parameters, plasma C-reactive protein (CRP), and tumor necrosis factor alpha were analyzed in 64 dogs with a presumptive diagnosis of pyometra. Final diagnosis (i.e., pyometra or cystic endometrial hyperplasia [CEH]) was determined by histopathology. As a single test, the percentage of band neutrophils had the highest sensitivity in the prediction of pyometra (sensitivity, 94%). The combination of percentage of bands and CRP had the highest sensitivity (97.7%; specificity, 75%) in predicting the presence of pyometra. The most common clinical signs noted in the study were vaginal discharge, polyuria, polydipsia, lethargy, and gastrointestinal signs. A combination of three or more of these clinical signs was significantly associated with pyometra. PMID- 15347620 TI - Efficacy of selamectin in the treatment of nasal mite (Pneumonyssoides caninum) infection in dogs. AB - In a laboratory study to evaluate the efficacy of selamectin for treatment of canine nasal mite infection, 12 purpose-bred beagles were experimentally infected with Pneumonyssoides caninum (P. caninum). Six of the dogs were treated with selamectin applied to the skin of the back at dosages of 6 to 24 mg/kg for three times at 2-week intervals. The remaining six dogs were an untreated control group. At necropsy 39 to 46 days after inoculation, no P. caninum mites were found in any of the treated dogs. In contrast, nasal mites were found in five of the untreated dogs. This difference was statistically significant at P=0.015. PMID- 15347621 TI - Investigation of Keyes skin biopsy instrument for intestinal biopsy versus a standard biopsy technique. AB - A prospective, clinical trial was conducted using 12 dogs that were presented for intestinal biopsy. Comparisons were made between paired jejunal biopsies collected using a Keyes biopsy punch and a standard scissor excisional technique. There were no differences in speed of collection, diagnostic value of specimens, complication rates, or sample quality. In this study, the Keyes skin punch was found to be safe, rapid, and diagnostic in all animals. Based on results of this study, the use of a Keyes punch instrument can be recommended as an alternative to other open intestinal biopsy methods. PMID- 15347622 TI - Pigmented epidermal plaques in three dogs. AB - Papillomavirus was identified in pigmented epidermal plaques (PEP) from three dogs: a miniature schnauzer with hyperadrenocorticism and hypoglobulinemia, an American Staffordshire terrier with hypoglobulinemia, and a Pomeranian with unconfirmed hypothyroidism. Squamous cell carcinoma (SCC) arose within several plaques in the Pomeranian. Clinical improvement coincided in the first two cases with treatment of the concurrent disease and the administration of low-dose oral interferon-alpha. This is the first report of PEP in an American Staffordshire terrier and a Pomeranian. The potential for malignant transformation of PEP to SCC emphasizes the need for recognition and clinical management of PEP. PMID- 15347623 TI - Group G streptococcal toxic shock-like syndrome in three cats. AB - Three 8-week-old kittens were presented with a history of acute, generalized weakness and severe fever. One cat was dead upon presentation, and necropsy findings were supportive of a group G Streptococcus spp. septicemia. During their clinical courses, two of the three kittens developed a progressive, marked swelling of one or more limbs. One moribund and severely hypothermic cat was euthanized a few hours after presentation, and necropsy was also supportive of a group G Streptococcus spp. septicemia. One kitten recovered. Group G streptococcal toxic shock-like syndrome was suspected because of the fulminant progression of the septicemia. PMID- 15347624 TI - Anemia associated with 'Candidatus Mycoplasma haemominutum' in a feline leukemia virus-negative cat with lymphoma. AB - 'Candidatus Mycoplasma haemominutum,' previously known as the small form of Haemobartonella felis (California species), is a hemotrophic parasite found on erythrocytes of infected cats. Although fleas are potential vectors, confirmatory studies are lacking. Healthy cats infected with 'Candidatus Mycoplasma haemominutum' generally do not have clinically significant anemia, but concurrent disease or immune suppression may predispose a cat to develop a life-threatening anemia, such as in the case reported here. PMID- 15347625 TI - Age differences in knee extension power, contractile velocity, and fatigability. AB - The purposes of this study were to examine age and gender differences in knee extensor strength, power, and fatigue using open- and closed-chain testing procedures. We tested the hypothesis that specific strength (strength/unit muscle mass) would not differ by age, whereas age differences in specific power and fatigue would remain consequent to blunted maximal contractile velocity. Skeletal muscle performance was examined in 28 young (26.9 +/- 0.7 yr) and 24 older (63.6 +/- 0.8 yr) men and women. Assessments included one-repetition maximum strength for knee extension, leg press, and squat; concentric knee extensor peak power, velocity, and fatigability; and sit-to-stand power, fatigability, and relative neural activation (electromyograph activity during sit-to-stand movement normalized to electromyograph activity during isometric maximum voluntary contraction). Thigh lean mass (TLM; kg) was assessed by dual-energy X-ray absorptiometry. Specific strength (N/kg TLM) and specific power (W/kg TLM) were estimated by dividing absolute values by TLM. Age differences in specific strength were observed for knee extension only (young, 41.2 +/- 1.0 N/kg TLM; older, 32.4 +/- 1.0 N/kg TLM; P < 0.05). Adjustment for TLM did not negate age differences in knee extension specific power (25-41% lower in older; P < 0.05) across loads tested. Older adults experienced fatigue across 10 repetitions of knee extension as peak velocity fell by 24% (P < 0.05). Deficits in concentric power persist after adjustment for TLM as maximum contractile velocity falls markedly with aging. Older adults are less capable of sustaining maximum concentric velocity during repetitive contractions. These findings suggest that velocity impairments are a possible contributor to mobility loss and falls risk among older adults. Interventions for improving contractile velocity should be pursued. PMID- 15347626 TI - Exercise training increases electron and substrate shuttling proteins in muscle of overweight men and women with the metabolic syndrome. AB - Aerobic conditioned muscle shows increased oxidative metabolism or glucose relative to untrained muscle at a given absolute exercise intensity. The studies of a targeted risk reduction intervention through defined exercise (STRRIDE) study is an aerobic exercise intervention in men and women with features of metabolic syndrome (Kraus WE, Torgan CE, Duscha BD, Norris J, Brown SA, Cobb FR, Bales CW, Annex BH, Samsa GP, Houmard JA, and Slentz CA, Med Sci Sports Exerc 33: 1774-1784, 2001), with four muscle biopsies taken during training and detraining time points. Here, we expanded a previous study (Hittel DS, Kraus WE, and Hoffman EP, J Physiol 548: 401-410, 2003) and used mRNA profiling to investigate gene transcripts associated with energy and substrate metabolism in STRRIDE participants. We found coordinate regulation of key metabolic enzymes with aerobic training in metabolic syndrome (aspartate aminotransferase 1, lactate dehydrogenase B, and pyruvate dehydrogenase-alpha(1)). All were also quickly downregulated by detraining, although the induction was not an acute response to activity. Protein and enzymatic assays were used to validate mRNA induction with aerobic training and loss with detraining (96 h to 2 wk) in 10 male and 10 female STRRIDE subjects. We propose that training coordinately increases the levels of aspartate aminotransferase 1, lactate dehydrogenase B, and pyruvate dehydrogenase alpha(1) subunit, increasing glucose metabolism in muscle by liberating pyruvate for oxidative metabolism and, therefore, limiting lactate efflux. Serial measurement of fasting plasma lactate from 62 subjects from the same exercise group demonstrated a significant decrease of circulating lactate with training. We also found evidence for sex-specific molecular remodeling of muscle with ubiquinol-cytochrome c reductase core protein II, a component of mitochondrial respiratory complex III, which showed an increase after training that was specific to women. These biochemical adaptations complement existing molecular models for improved glucose tolerance with exercise intervention in prediabetic individuals. PMID- 15347627 TI - Smooth muscle development during postnatal growth of distal bronchioles in infant rhesus monkeys. AB - Development of smooth muscle in conducting airways begins early in fetal life. Whereas the pattern and regulation of smooth muscle differentiation are well defined, the impact of airway growth on the process is not. To evaluate the transformations in organization during postnatal growth, smooth muscle bundle organization (size, abundance, and orientation) was mapped in five generations of distal airways of infant rhesus monkeys (5 days and 1, 2, 3, and 6 mo old). On the basis of direct measurement of the bronchiole proximal to the terminal bronchiole, length increased by 2-fold, diameter by 1.35-fold, and surface area by 2.8-fold between 5 days and 6 mo of age. Smooth muscle bundle size was greater in proximal bronchioles than in respiratory bronchioles and did not change with age. However, relative bundle size decreased in proportion to airway size as the airways grew. Relative bundle abundance was constant regardless of airway generation or age. The distribution of smooth muscle bundle orientation changed with age in each airway generation, and there were significant changes in the terminal and respiratory bronchioles. We conclude that smooth muscle undergoes marked organizational changes as airways grow during postnatal development. PMID- 15347628 TI - ACL deficiency impairs the vasoconstrictive efficacy of neuropeptide Y and phenylephrine in articular tissues: a laser speckle perfusion imaging study. AB - Sympathetic-derived neuropeptide Y (NPY) helps regulate inflammatory responses in injury and disease, is a vasoconstrictor, and stimulates angiogenesis. Rupture of the anterior cruciate ligament (ACL) is a common clinical presentation that results in tissue inflammation, hyperemia, and angiogenesis in the intact medial collateral ligament (MCL). This study is the first to examine the vasoregulatory role of NPY in ACL-deficient knee joints by using the newly developed technique of laser speckle perfusion imaging (LSPI). MCL blood flow was measured in two groups of adult rabbits: unoperated control (n = 6), and 6-wk ACL transected (n = 5). Under anesthesia, the MCL was surgically exposed and tissue blood flow was imaged at high resolution using LSPI. NPY was applied to the MCL vasculature in topical boluses of 100 mul (dose range 10(-14) to 10(-9) mol), and the alpha adrenoceptor agonist phenylephrine was applied in doses of 10(-14), 10(-10), and 10(-7) mol. In control rabbits, topical administration of NPY or phenylephrine produced dose-dependent vasopressor responses (maximal effect at 10(-9) mol NPY and 10(-7) mol phenylephrine). In ACL-transected knees, there was little or no vasoconstrictive response to NPY at any dose. The response to phenylephrine was significantly reduced compared with control ligaments. Possible causes of the reduced vasoconstrictive response to NPY in the MCL after 6 wk of ACL deficiency include development of tolerance to the peptide due to a prolonged increase in sympathetic nerve activity or change in the distribution or functionality of the NPY Y(1) receptors. Chronic ACL deficiency leads to profound and protracted hyperemia in associated articular tissues. Abrogation of a vasoconstrictor response to both NPY and phenylephrine in the MCL indicates that ACL deficiency induces major changes in the vascular physiological homeostasis. PMID- 15347629 TI - Age and regional specificity of peak limb vascular conductance in men. AB - Because of methodological variation in previous studies, age-associated changes in peak limb vascular conductance (VC(peak); a functional index of arterial structure) and its determinants remain poorly defined. The objectives of this study were to describe and compare age-associated changes in peak forearm and calf conductance across a broad age range and to identify physiological characteristics that are predictive of variation in limb-specific VC(peak). Peak conductance (plethysmographic flow/brachial mean arterial pressure) of the forearm (forearm VC(peak)) and calf (calf VC(peak)) after 10 min of arterial occlusion was measured twice in 68 healthy, normally active men aged 20-79 yr. Aerobic capacity (cycle peak oxygen consumption), arterial health (ankle-brachial index, pulse wave velocity), and limb-specific measures of muscle mass (dual energy X-ray absorptiometry) and isometric strength (grip, plantar flexion) were also assessed. The relative decline in forearm VC(peak) with age (-6.6% per decade; P < 0.001) was greater than the decline in calf VC(peak) (-3.4% per decade; P = 0.004). Limb VC(peak) per kilogram of muscle declined with age in the forearm (-3.8% per decade; P = 0.004) but not in the calf (P = 0.35). Age, Vo(2 peak), and regional muscle mass were significant predictors of peak conductance in both limbs; however, these predictors explained considerably less variance in the calf than in the forearm. These results suggest that healthy aging is associated with a linear decline in limb vasodilator capacity in men, but the magnitude of this effect is reduced in the calf relative to the forearm. This could reflect regional differences in habitual muscle use with aging in normally active men. PMID- 15347630 TI - Tendon elongation influences the amplitude of interpolated doublets in the assessment of activation in elderly men. AB - This study investigated the influence of tendon elongation (TE) on postcontraction doublet (PCD) torque in the assessment of activation in the plantar flexors of nine elderly men (EM, age 73.7 +/- 3.6 yr) and nine young men (YM, age 24.7 +/- 4.7 yr). Plantar flexion maximal voluntary contractions (MVC) and activation were assessed at ankle joint angles of -20 degrees (dorsiflexion), 0 degrees , and 20 degrees (plantar flexion). Across the ankle joint angles tested, compared with YM, the EM had a 36-49% lower plantar flexion MVC (P < 0.01), TE was greater by 25-31% (P < 0.01), and electromechanical delay was 65 108% greater (P < 0.01). Activation (PCD torque to interpolated doublet torque) was 15% lower in EM compared with YM at -20 degrees (P < 0.05), but no different at 0 and 20 degrees . In the EM, PCD torque relative to MVC torque was significantly lower at 20 degrees compared with 0 degrees (P < 0.05). Electromechanical delay was positively correlated with TE (R(2) = 0.489, P < 0.01). In conclusion, this investigation demonstrates that, although a negative association exists between TE and PCD torque, the consequence of a greater TE on the estimation of activation in EM is negligible. This is due to a greater influence of ankle joint angle on the occlusion of a superimposed doublet, which counteracts the lesser influence of joint angle on TE and PCD torque. However, a greater TE in EM was found to significantly increase electromechanical delay, which is expected to influence the time needed for postural readjustments. PMID- 15347631 TI - Mucociliary and long-term particle clearance in the airways of healthy nonsmoker subjects. AB - Spherical monodisperse ferromagnetic iron oxide particles of 1.9-microm geometric and 4.2-microm aerodynamic diameter were inhaled by 13 healthy nonsmoking subjects using the shallow bolus technique. The bolus width was 100 ml, and the penetration front depth was 150 +/- 27 ml. The mean flow rate during inhalation and exhalation was 250 ml/s. The Fowler dead space and the phase 1 dead space of the airways were 282 +/- 49 and 164 +/- 34 ml, respectively. Deposition was below 20% without breath holding and 51 +/- 8% after an 8-s breath-holding time. We attempted to confine the bolus deposition to the bronchial airways by limiting the bolus front depth to the phase 1 dead space volume. Particle retention was measured by the magnetopneumographic method over a period of 9 mo. Particle clearance from the airways showed a fast and a slow phase; 49 +/- 9% followed the fast phase with a mean half-time of 3.0 +/- 1.6 h and characterized the mucociliary clearance. The remaining fraction was cleared slowly with a half-time of 109 +/- 78 days. The slow clearance phase was comparable to clearance measurements from the lung periphery of healthy nonsmokers, which allowed macrophage-dependent clearance mechanisms of the slow cleared fraction to be taken into account. Despite the fact that part of the slowly cleared particles may originate from peripheral deposition, the data demonstrate that mucociliary clearance does not remove all particles deposited in the airways and that a significant fraction undergoes long-term retention mechanisms, the origin of which is still under discussion. PMID- 15347632 TI - Relationship between muscle blood flow and oxygen uptake during exercise in endurance-trained and untrained men. AB - A recent study showed good correlation between regional blood flow (BF) and oxygen uptake (Vo(2)) 30 min after exhaustive exercise. The question that remains open is whether there is similar good correlation between BF and Vo(2) also during exercise. We reanalyzed our previous data from a study in which BF and Vo(2) was measured in different quadriceps femoris muscles in seven healthy endurance-trained and seven healthy untrained men at rest and during low intensity intermittent static knee-extension exercise (Kalliokoski KK, Oikonen V, Takala TO, Sipila H, Knuuti J, and Nuutila P. Am J Physiol Endocrinol Metab 280: E1015-E1021, 2001). When the mean values of each muscle were considered, there was good correlation between BF and Vo(2) during exercise in both groups (r(2) = 0.82 in untrained and 0.97 in trained). However, when calculated individually, the correlations were poorer, and the mean correlation coefficient (r(2)) was significantly higher in the trained men (0.71 +/- 0.07 vs. 0.40 +/- 0.11, P = 0.03). These results suggest that there is large individual variation in matching BF to Vo(2) in human skeletal muscles during exercise, ranging from very poor to excellent. Furthermore, this matching seems to be better in the endurance-trained than in untrained men. PMID- 15347633 TI - Variability and variation in drug susceptibility among Giardia duodenalis isolates and clones exposed to 5-nitroimidazoles and benzimidazoles in vitro. AB - OBJECTIVES: We analysed, in a cell-by-cell study, the in vitro susceptibility of Giardia duodenalis strains, including Mexican isolates and their clones to 5 nitroimidazoles and benzimidazoles. METHODS: Fluorogenic dye staining (FDA-PI) and cell morphology (CM) assays, two fast and direct techniques, replaced the indirect 'gold standard' method (subculture in liquid medium) in the evaluation of 5-nitroimidazoles and benzimidazoles, respectively. RESULTS: Under these conditions, the activity of several 5-nitroimidazole and benzimidazole derivatives was consistent with their known efficacy, but parasite stocks exhibited a greater variability in response to 5-nitroimidazoles compared with benzimidazoles. Also, consecutive progenies from single stocks maintained in continuous subculture in drug-free media displayed changes (variations) in the proportions of drug resistant (R/T) subpopulations when exposed to sublethal concentrations of 5-nitroimidazoles and benzimidazoles. These were again more variable upon exposure to 5-nitroimidazoles than to benzimidazoles. Variations were not due to drug susceptibility shifts in parent trophozoites since analysis of cytokinetic processes showed a predominant pattern of susceptible/susceptible or resistant/resistant daughters, whereas susceptible/resistant daughters were scarce. CONCLUSIONS: Our observations support the idea that G. duodenalis cultures exhibit variations in their response to 5-nitroimidazoles and benzimidazoles as a result of a drug-independent competition between drug susceptible and drug-resistant subpopulations when parasites are subcultured. PMID- 15347634 TI - Comparison of short-term treatment regimen of ciprofloxacin versus long-term treatment regimens of trimethoprim/sulfamethoxazole or norfloxacin for uncomplicated lower urinary tract infections: a randomized, multicentre, open label, prospective study. AB - OBJECTIVE: To compare the bacteriological and clinical efficacy of three treatments for uncomplicated cystitis in ambulatory pre-menopausal women: ciprofloxacin 250 mg orally twice daily for 3 days, trimethoprim/sulfamethoxazole 160/800 mg orally twice daily for 7 days, and norfloxacin 400 mg orally twice daily for 7 days. MATERIALS AND METHODS: A total of 455 women were randomly assigned to three treatment groups: 151 received ciprofloxacin, 150 received trimethoprim/sulfamethoxazole, and 154 received norfloxacin. Bacteriological cure and clinical resolution were evaluated 5-9 days and 4-6 weeks after completion of treatment. RESULTS: There was no significant difference among the three treatment groups: overall efficacy ranged from 78.5% for the trimethoprim/sulfamethoxazole group, to 84.5% for the ciprofloxacin group. The highest overall incidence of drug-related adverse effects occurred in the trimethoprim/sulfamethoxazole patients. CONCLUSIONS: These data indicate that a 3 day treatment with ciprofloxacin is at least as clinically and bacteriologically effective as 7 day treatments with trimethoprim/sulfamethoxazole and norfloxacin for uncomplicated lower urinary tract infections. PMID- 15347635 TI - Activity of capuramycin analogues against Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium intracellulare in vitro and in vivo. AB - OBJECTIVES: The antimycobacterial activities of RS-112997, RS-124922 and RS 118641, three capuramycin analogues that inhibit phospho-N-acetylmuramyl pentapeptide translocase, were tested against clinical isolates of Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium intracellulare. METHODS AND RESULTS: MICs were determined by the broth microdilution method using a modified Middlebrook 7H9 broth. RS-118641 was the most potent compound overall. The MIC50/90 (mg/L) results for RS-118641 were: M. tuberculosis, 1/2; multidrug resistant (MDR) M. tuberculosis, 0.5/2; M. avium, 4/8; and M. intracellulare, 0.06/0.5. No statistically significant differences in MIC distributions were observed between non-MDR and MDR M. tuberculosis for any of the capuramycin analogues tested. In order to evaluate the therapeutic efficacy of RS-112997 and RS-124922 in a murine lung model of tuberculosis, both compounds were administered intranasally at 0.1 or 1 mg/mouse/day for 12 days. The mycobacterial load in the lungs was significantly lower in all treatment groups than in the untreated controls. Additional experiments were performed to evaluate the therapeutic efficacy of the three compounds against the M. intracellulare infection in mice. All compounds were administered intranasally at 0.1 mg/mouse/day for 21 days. The mycobacterial load in the lungs was significantly lower in all treatment groups than in the untreated controls. CONCLUSIONS: These results suggest that capuramycin analogues exhibit strong antimycobacterial potential and should be considered for further evaluation in the treatment of M. tuberculosis and M. avium-M. intracellulare complex infections in humans. PMID- 15347636 TI - Pharmacokinetics of moxifloxacin in patients undergoing continuous venovenous haemodiafiltration. AB - OBJECTIVE: Moxifloxacin is an 8-methoxy quinolone with a broad range of activity against clinically important pathogens. Therefore it is frequently administered in severe respiratory tract infections. Continuous venovenous haemodiafiltration (CVVHDF) is an important extracorporeal renal replacement therapy for intensive care patients suffering from sepsis and multiple organ failure. The aim of this study was to investigate the pharmacokinetics of intravenous moxifloxacin in anuric critically ill patients undergoing CVVHDF. PATIENTS AND METHODS: Pharmacokinetic analysis was performed in nine intensive care patients with acute renal failure and suspected or proven infection sensitive to moxifloxacin, who received moxifloxacin 400 mg intravenously once daily. The concentration of moxifloxacin in serum and ultradiafiltrate was determined by HPLC. RESULTS: Peak and trough serum concentrations were 3.76 +/- 2.02 mg/L and 0.24 +/- 0.14 mg/L, respectively, at the arterial port after the first dose. The mean elimination half-life was 9.87 +/- 3.26 h, the volume of distribution 270 +/- 133 L and the calculated AUC0-24 18.41 +/- 8.46 mg.h/L. Total clearance was 19.09 +/- 8.22 L/h and the clearance of haemodiafiltration 1.63 +/- 0.33 L/h. CONCLUSIONS: The pharmacokinetics of moxifloxacin in critically ill patients with acute renal failure undergoing CVVHDF was comparable to healthy subjects and patients without renal impairment. We recommend 400 mg of intravenous moxifloxacin once per day in anuric patients during CVVHDF. PMID- 15347637 TI - Reversion to susceptibility in a linezolid-resistant clinical isolate of Staphylococcus aureus. AB - OBJECTIVES: Linezolid resistance in rare isolates of Staphylococcus aureus has been associated with G2576T mutations in domain V of the 23S rRNA gene. We report the analysis of a clinical S. aureus isolate that developed linezolid resistance (MIC of linezolid of 12 mg/L) after a 25 day course of the drug. Sequencing identified G2576T mutations in four of the five copies of the 23S rRNA gene. METHODS: To examine the stability of this resistance, we serially passaged this original isolate 60 times over a 75 day period on antimicrobial-free medium. RESULTS: After 30 passages, the MIC of linezolid had decreased to 8 mg/L and only two of the five copies of the 23S rRNA gene contained the G2576T mutation. After 60 passages, the MIC of linezolid fell to 2 mg/L and only one of the five 23S rRNA gene copies contained the mutation. The original and two passaged staphylococci were indistinguishable by pulsed-field gel electrophoresis. CONCLUSIONS: In the absence of antibiotic pressure, linezolid resistance was unstable in a clinical isolate that did not have all copies of the 23S rRNA gene mutated, although a single copy of mutant rDNA was maintained. Gene conversion was probably the mechanism for this reversion, using the wild-type 23S rRNA gene sequences to replace the G2576T mutation by homologous recombination. PMID- 15347638 TI - Community and hospital spread of Escherichia coli producing CTX-M extended spectrum beta-lactamases in the UK. AB - OBJECTIVES: During 2003, the Health Protection Agency's Antibiotic Resistance Monitoring and Reference Laboratory began to receive isolates of Escherichia coli for confirmation of extended-spectrum beta-lactamase production with a phenotype implying a CTX-M-type beta-lactamase, i.e. MICs of cefotaxime > or = 8-fold higher than MICs of ceftazidime. Many were referred as being from community patients. We examined 291 CTX-M-producing isolates from the UK and investigated the genetic basis of their phenotype. METHODS: PCR was used to detect alleles encoding CTX-M enzymes and to assign these to their blaCTX-M phylogenetic groups. Selected alleles were sequenced. Producers were compared by analysis of banding patterns generated by pulsed-field gel electrophoresis of XbaI-digested genomic DNA. MICs were determined by an agar dilution method or by Etest. RESULTS: Of 291 CTX-M-producing E. coli isolates studied from 42 UK centres, 70 (24%) were reportedly from community patients, many of whom had only limited recent hospital contact. Community isolates were referred by 12 centres. Two hundred and seventy nine (95.9%) producers contained genes encoding group 1 CTX-M enzymes and 12 contained blaCTX-M-9-like alleles. An epidemic CTX-M-15-producing strain was identified, with 110 community and inpatient isolates referred from six centres. Representatives of four other major strains also produced CTX-M-15, as did several sporadic isolates examined. Most producers were multi-resistant to fluoroquinolones, trimethoprim, tetracycline and aminoglycosides as well as to non-carbapenem beta-lactams. CONCLUSIONS: CTX-M-producing E. coli are a rapidly developing problem in the UK, with CTX-M-15 particularly common. The diversity of producers and geographical scatter of referring laboratories indicates wide dissemination of blaCTX-M genes. Because of the public health implications, including for the treatment of community-acquired urinary tract infections, the spread of these strains--and CTX-M-15 beta-lactamase in particular--merits close monitoring. PMID- 15347639 TI - Genetic relatedness of ciprofloxacin-resistant Shigella dysenteriae type 1 strains isolated in south Asia. AB - OBJECTIVES: The aim of the present study was to determine the clonal relationships of ciprofloxacin-resistant Shigella dysenteriae type 1 strains isolated from south Asia, and S. dysenteriae 1 strains associated with epidemics in 1978, 1984 and 1994. METHODS: The antimicrobial susceptibilities were examined by NCCLS methods. Molecular epidemiological characterization was performed by plasmid profiling, pulsed-field gel electrophoresis (PFGE) and mutation analysis of the quinolone resistance-determining region (QRDR) of gyrA by sequencing. RESULTS: Plasmid patterns of the current ciprofloxacin-resistant strains from India, Nepal and Bangladesh were very similar to those of the 1978, 1984 and 1994 epidemic isolates of S. dysenteriae 1, except for the presence of a new plasmid of approximately 2.6 MDa, which was found in one recent ciprofloxacin-resistant strain isolated in Bangladesh. PFGE analysis showed that the ciprofloxacin resistant strains isolated in Bangladesh, India and Nepal belonged to a PFGE type (type A), which was possibly related to that of the 1984 and 1994 clone of S. dysenteriae 1, but different from 1978 epidemic strains. The current ciprofloxacin-resistant strains belong to five subtypes (A3-A7), all of which were found in India, but in Bangladesh and Nepal, only A3 existed. Mutation analysis of the QRDR of gyrA revealed that amino acid substitutions at positions 83 and 87 of ciprofloxacin-resistant strains isolated in Bangladesh were similar to those of the strains isolated in Nepal, but different (at position 87) from ciprofloxacin-resistant strains isolated in India. CONCLUSIONS: PFGE and mutation analysis of gyrA showed differences between the current ciprofloxacin-resistant S. dysenteriae 1 strains isolated in south Asia and those associated with epidemics in 1978, 1984 and 1994. PMID- 15347640 TI - Bacterial long chain fatty acid transport: gateway to a fatty acid-responsive signaling system. PMID- 15347641 TI - Electrostatic interaction between redox cofactors in photosynthetic reaction centers. AB - Intramolecular electron transfer within proteins is an essential process in bioenergetics. Redox cofactors are embedded in proteins, and this matrix strongly influences their redox potential. Several cofactors are usually found in these complexes, and they are structurally organized in a chain with distances between the electron donor and acceptor short enough to allow rapid electron tunneling. Among the different interactions that contribute to the determination of the redox potential of these cofactors, electrostatic interactions are important but restive to direct experimental characterization. The influence of interaction between cofactors is evidenced here experimentally by means of redox titrations and time-resolved spectroscopy in a chimeric bacterial reaction center (Maki, H., Matsuura, K., Shimada, K., and Nagashima, K. V. P. (2003) J. Biol. Chem. 278, 3921-3928) composed of the core subunits of Rubrivivax gelatinosus and the tetraheme cytochrome of Blastochloris viridis. The absorption spectra and orientations of the various cofactors of this chimeric reaction center are similar to those found in their respective native protein, indicating that their local environment is conserved. However, the redox potentials of both the primary electron donor and its closest heme are changed. The redox potential of the primary electron donor is downshifted in the chimeric reaction center when compared with the wild type, whereas, conversely, that of its closet heme is upshifted. We propose a model in which these reciprocal shifts in the midpoint potentials of two electron transfer partners are explained by an electrostatic interaction between them. PMID- 15347642 TI - Modulation of methyl group metabolism by streptozotocin-induced diabetes and all trans-retinoic acid. AB - The hepatic enzyme glycine N-methyltransferase (GNMT) plays a major role in the control of methyl group and homocysteine metabolism. Because disruption of these vital pathways is associated with numerous pathologies, understanding GNMT control is important for evaluating methyl group regulation. Recently, gluconeogenic conditions have been shown to modulate homocysteine metabolism and treatment with glucocorticoids and/or all-trans-retinoic acid (RA)-induced active GNMT protein, thereby leading to methyl group loss. This study was conducted to determine the effect of diabetes, alone and in combination with RA, on GNMT regulation. Diabetes and RA increased GNMT activity 87 and 148%, respectively. Moreover, the induction of GNMT activity by diabetes and RA was reflected in its abundance. Cell culture studies demonstrated that pretreatment with insulin prevented GNMT induction by both RA and dexamethasone. There was a significant decline in homocysteine concentrations in diabetic rats, owing in part to a 38% increase in the abundance of the transsulfuration enzyme cystathionine beta synthase; treatment of diabetic rats with RA prevented cystathionine beta synthase induction. A diabetic state also increased the activity of the folate independent homocysteine remethylation enzyme betaine-homocysteine S methyltransferase, whereas the activity of the folate-dependent enzyme methionine synthase was diminished 52%. In contrast, RA treatment attenuated the streptozotocin-mediated increase in betaine-homocysteine S-methyltransferase, whereas methionine synthase activity remained diminished. These results indicate that both a diabetic condition and RA treatment have marked effects on the metabolism of methyl groups and homocysteine, a finding that may have significant implications for diabetics and their potential sensitivity to retinoids. PMID- 15347643 TI - Non-muscle myosin II and myosin light chain kinase are downstream targets for vasopressin signaling in the renal collecting duct. AB - We have previously demonstrated that vasopressin increases the water permeability of the inner medullary collecting duct (IMCD) by inducing trafficking of aquaporin-2 to the apical plasma membrane and that this response is dependent on intracellular calcium mobilization and calmodulin activation. Here, we address the hypothesis that this water permeability response is mediated in part through activation of the calcium/calmodulin-dependent myosin light chain kinase (MLCK) and regulation of non-muscle myosin II. Immunoblotting and immunocytochemistry demonstrated the presence of MLCK, the myosin regulatory light chain (MLC), and the IIA and IIB isoforms of the non-muscle myosin heavy chain in rat IMCD cells. Two-dimensional electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry identified two isoforms of MLC, both of which also exist in phosphorylated and non-phosphorylated forms. 32P incubation of the inner medulla followed by autoradiography of two-dimensional gels demonstrated increased 32P labeling of both isoforms in response to the V2 receptor agonist [deamino-Cys1,D-Arg8]vasopressin (DDAVP). Time course studies of MLC phosphorylation in IMCD suspensions (using immunoblotting with anti-phospho-MLC antibodies) showed that the increase in phosphorylation could be detected as early as 30 s after exposure to vasopressin. The MLCK inhibitor ML-7 blocked the DDAVP-induced MLC phosphorylation and substantially reduced [Arg8]vasopressin (AVP)-stimulated water permeability. AVP-induced MLC phosphorylation was associated with a rearrangement of actin filaments (Alexa Fluor 568-phalloidin) in primary cultures of IMCD cells. These results demonstrate that MLC phosphorylation by MLCK represents a downstream effect of AVP-activated calcium/calmodulin signaling in IMCD cells and point to a role for non-muscle myosin II in regulation of water permeability by vasopressin. PMID- 15347644 TI - Glutaredoxin 2 catalyzes the reversible oxidation and glutathionylation of mitochondrial membrane thiol proteins: implications for mitochondrial redox regulation and antioxidant DEFENSE. AB - The redox poise of the mitochondrial glutathione pool is central in the response of mitochondria to oxidative damage and redox signaling, but the mechanisms are uncertain. One possibility is that the oxidation of glutathione (GSH) to glutathione disulfide (GSSG) and the consequent change in the GSH/GSSG ratio causes protein thiols to change their redox state, enabling protein function to respond reversibly to redox signals and oxidative damage. However, little is known about the interplay between the mitochondrial glutathione pool and protein thiols. Therefore we investigated how physiological GSH/GSSG ratios affected the redox state of mitochondrial membrane protein thiols. Exposure to oxidized GSH/GSSG ratios led to the reversible oxidation of reactive protein thiols by thiol-disulfide exchange, the extent of which was dependent on the GSH/GSSG ratio. There was an initial rapid phase of protein thiol oxidation, followed by gradual oxidation over 30 min. A large number of mitochondrial proteins contain reactive thiols and most of these formed intraprotein disulfides upon oxidation by GSSG; however, a small number formed persistent mixed disulfides with glutathione. Both protein disulfide formation and glutathionylation were catalyzed by the mitochondrial thiol transferase glutaredoxin 2 (Grx2), as were protein deglutathionylation and the reduction of protein disulfides by GSH. Complex I was the most prominent protein that was persistently glutathionylated by GSSG in the presence of Grx2. Maintenance of complex I with an oxidized GSH/GSSG ratio led to a dramatic loss of activity, suggesting that oxidation of the mitochondrial glutathione pool may contribute to the selective complex I inactivation seen in Parkinson's disease. Most significantly, Grx2 catalyzed reversible protein glutathionylation/deglutathionylation over a wide range of GSH/GSSG ratios, from the reduced levels accessible under redox signaling to oxidized ratios only found under severe oxidative stress. Our findings indicate that Grx2 plays a central role in the response of mitochondria to both redox signals and oxidative stress by facilitating the interplay between the mitochondrial glutathione pool and protein thiols. PMID- 15347645 TI - Differential osteopontin expression in phenotypically distinct subclones of murine breast cancer cells mediates metastatic behavior. AB - Cancer progression depends on an accumulation of metastasis-supporting cell signaling molecules, which target signal transduction pathways and, ultimately, gene expression. One such molecule, osteopontin (OPN), represents a key molecular signaling event in tumor progression and metastasis. However, the transcriptional regulatory mechanisms that underlie OPN expression in the setting of breast cancer have not been well studied. In this regard, we have examined the differential transcriptional regulation of OPN in the murine mammary epithelial tumor cell lines, 4T1 and 4T07, which are sublines derived from the parental population of 410.4 cells from Balb/cfC3H mice. These lines are phenotypically heterogeneous in their metastatic behavior. 4T1 hematogenously metastasizes to the lung, liver, bone, and brain, whereas 4T07 is highly tumorigenic but fails to metastasize. The tumor growth and metastatic spread of 4T1 cells closely mimics stage IV breast cancer. We demonstrate that a Ras-independent, phosphoinositide-3 kinase-dependent, c-Jun N-terminal kinase-dependent phosphorylation of c-Jun results in binding of an AP-1 c-Jun homodimer to the OPN promoter in 4T1 cells. This differential up-regulation of OPN gene transcription and protein expression in 4T1 cells conveys in vitro correlates of a metastatic phenotype. These results provide new insight into the transcriptional regulation of OPN as a key mediator of metastatic behavior in malignancy. PMID- 15347646 TI - The Leber congenital amaurosis protein AIPL1 modulates the nuclear translocation of NUB1 and suppresses inclusion formation by NUB1 fragments. AB - Mutations in the aryl hydrocarbon receptor-interacting protein-like 1 (AIPL1) cause the blinding disease Leber congenital amaurosis (LCA). The similarity of AIPL1 to AIP has led to suggestions that AIPL1 could function in a similar manner to AIP in facilitating protein translocation and as a component of chaperone complexes. AIPL1 interacts with the cell cycle regulator NEDD8 ultimate buster protein 1 (NUB1). As AIPL1 is predominantly cytoplasmic and NUB1 is predominantly nuclear, we tested the hypothesis that AIPL1 could modulate the nuclear translocation of NUB1. Co-transfection of AIPL1 with GFP-NUB1 resulted in a shift of GFP-NUB1 subcellular distribution toward the cytoplasm. Interestingly, AIPL1 was able to act in a chaperone-like fashion to efficiently suppress inclusion formation by NUB1 fragments. Co-transfection of AIPL1 with GFP-NUB1-N and GFP NUB1-C resulted in an AIPL1-dependent suppression of GFP-NUB1-N perinuclear inclusions and GFP-NUB1-C intranuclear inclusions leading to the redistribution of these fragments in the cytoplasm. This chaperone-like function of AIPL1 was specific for NUB1, since AIPL1 was unable to suppress the inclusion formation by unrelated aggregation-prone proteins and AIP had no effect on NUB1 localization or inclusion formation. We examined the effect of a range of pathogenic and engineered mutations on the ability of AIPL1 to modulate NUB1 localization or inclusion formation. With the exception of W278X, which formed non-functional SDS insoluble inclusions, all of the pathogenic mutations studied were soluble and could modulate NUB1 with varying efficiency compared with the wild-type protein. The effect of AIPL1 on NUB1 required the C-terminal region of AIPL1, as engineered C-terminal truncation mutations had no effect on NUB1. These data show that AIPL1 can modulate protein translocation and act in a chaperone-like manner and suggest that AIPL1 is an important modulator of NUB1 cellular function. PMID- 15347647 TI - Active site engineering of the epoxide hydrolase from Agrobacterium radiobacter AD1 to enhance aerobic mineralization of cis-1,2-dichloroethylene in cells expressing an evolved toluene ortho-monooxygenase. AB - Chlorinated ethenes are the most prevalent ground-water pollutants, and the toxic epoxides generated during their aerobic biodegradation limit the extent of transformation. Hydrolysis of the toxic epoxide by epoxide hydrolases represents the major biological detoxification strategy; however, chlorinated epoxyethanes are not accepted by known bacterial epoxide hydrolases. Here, the epoxide hydrolase from Agrobacterium radiobacter AD1 (EchA), which enables growth on epichlorohydrin, was tuned to accept cis-1,2-dichloroepoxyethane as a substrate by accumulating beneficial mutations from three rounds of saturation mutagenesis at three selected active site residues, Phe-108, Ile-219, and Cys-248 (no beneficial mutations were found at position Ile-111). The EchA F108L/I219L/C248I variant coexpressed with a DNA-shuffled toluene ortho-monooxygenase, which initiates attack on the chlorinated ethene, enhanced the degradation of cis dichloroethylene (cis-DCE) an infinite extent compared with wild-type EchA at low concentrations (6.8 microm) and up to 10-fold at high concentrations (540 microm). EchA variants with single mutations (F108L, I219F, or C248I) enhanced cis-DCE mineralization 2.5-fold (540 microm), and EchA variants with double mutations, I219L/C248I and F108L/C248I, increased cis-DCE mineralization 4- and 7 fold, respectively (540 microm). For complete degradation of cis-DCE to chloride ions, the apparent Vmax/Km for the Escherichia coli strain expressing recombinant the EchA F108L/I219L/C248I variant was increased over 5-fold as a result of the evolution of EchA. The EchA F108L/I219L/C248I variant also had enhanced activity for 1,2-epoxyhexane (2-fold) and the natural substrate epichlorohydrin (6-fold). PMID- 15347648 TI - Characterization of the menaquinone-dependent disulfide bond formation pathway of Escherichia coli. AB - In the protein disulfide-introducing system of Escherichia coli, plasma membrane integrated DsbB oxidizes periplasmic DsbA, the primary disulfide donor. Whereas the DsbA-DsbB system utilizes the oxidizing power of ubiquinone (UQ) under aerobic conditions, menaquinone (MK) is believed to function as an immediate electron acceptor under anaerobic conditions. Here, we characterized MK reactivities with DsbB. In the absence of UQ, DsbB was complexed with MK8 in the cell. In vitro studies showed that, by binding to DsbB in a manner competitive with UQ, MK specifically oxidized Cys41 and Cys44 of DsbB and activated its catalytic function to oxidize reduced DsbA. In contrast, menadione used in earlier studies proved to be a more nonspecific oxidant of DsbB. During catalysis, MK8 underwent a spectroscopic transition to develop a visible violet color (lambdamax = 550 nm), which required a reduced state of Cys44 as shown previously for UQ color development (lambdamax = 500 nm) on DsbB. In an in vitro reaction system of MK8-dependent oxidation of DsbA at 30 degrees C, two reaction components were observed, one completing within minutes and the other taking >1 h. Both of these reaction modes were accompanied by the transition state of MK, for which the slower reaction proceeded through the disulfide-linked DsbA DsbB(MK) intermediate. The MK-dependent pathway provides opportunities to further dissect the quinone-dependent DsbA-DsbB redox reactions. PMID- 15347649 TI - Novel phenotypes of Escherichia coli tat mutants revealed by global gene expression and phenotypic analysis. AB - The Tat protein export system serves to export folded proteins harboring an N terminal twin arginine signal peptide across the cytoplasmic membrane. In this study, we have used gene expression profiling of Escherichia coli supported by phenotypic analysis to investigate how cells respond to a defect in the Tat pathway. Previous work has demonstrated that strains mutated in genes encoding essential Tat pathway components are defective in the integrity of their cell envelope because of the mislocalization of two amidases involved in cell wall metabolism (Ize, B., Stanley, N. R., Buchanan, G., and Palmer, T. (2003) Mol. Microbiol. 48, 1183-1193). To distinguish between genes that are differentially expressed specifically because of the cell envelope defect and those that result from other effects of the tatC deletion, we also analyzed two different transposon mutants of the DeltatatC strain that have their outer membrane integrity restored. Approximately 50% of the genes that were differentially expressed in the tatC mutant are linked to the envelope defect, with the products of many of these genes involved in self-defense or protection mechanisms, including the production of exopolysaccharide. Among the changes that were not explicitly linked to envelope integrity, we characterized a role for the Tat system in iron acquisition and copper homeostasis. Finally, we have demonstrated that overproduction of the Tat substrate SufI saturates the Tat translocon and produces effects on global gene expression that are similar to those resulting from the DeltatatC mutation. PMID- 15347650 TI - BeF(x) stops the chaperonin cycle of GroEL-GroES and generates a complex with double folding chambers. AB - Coupling with ATP hydrolysis and cooperating with GroES, the double ring chaperonin GroEL assists the folding of other proteins. Here we report novel GroEL-GroES complexes formed in fluoroberyllate (BeF(x)) that can mimic the phosphate part of the enzyme-bound nucleotides. In ATP, BeF(x) stops the functional turnover of GroEL by preventing GroES release and produces a symmetric 1:2 GroEL-GroES complex in which both GroEL rings contain ADP.BeF(x) and an encapsulated substrate protein. In ADP, the substrate protein-loaded GroEL cannot bind GroES. In ADP plus BeF(x), however, it can bind GroES to form a stable 1:1 GroEL-GroES complex in which one of GroEL rings contains ADP.BeF(x) and an encapsulated substrate protein. This 1:1 GroEL-GroES complex is converted into the symmetric 1:2 GroEL-GroES complex when GroES is supplied in ATP plus BeF(x). Thus, BeF(x) stabilizes two GroEL-GroES complexes; one with a single folding chamber and the other with double folding chambers. These results shed light on the intermediate ADP.P(i) nucleotide states in the functional cycle of GroEL. PMID- 15347651 TI - Differential expression and interaction with the visual G-protein transducin of centrin isoforms in mammalian photoreceptor cells. AB - Photoisomerization of rhodopsin activates a heterotrimeric G-protein cascade leading to closure of cGMP-gated channels and hyperpolarization of photoreceptor cells. Massive translocation of the visual G-protein transducin, Gt, between subcellular compartments contributes to long term adaptation of photoreceptor cells. Ca(2+)-triggered assembly of a centrin-transducin complex in the connecting cilium of photoreceptor cells may regulate these transducin translocations. Here we demonstrate expression of all four known, closely related centrin isoforms in the mammalian retina. Interaction assays revealed binding potential of the four centrin isoforms to Gtbetagamma heterodimers. High affinity binding to Gtbetagamma and subcellular localization of the centrin isoforms Cen1 and Cen2 in the connecting cilium indicated that these isoforms contribute to the centrin-transducin complex and potentially participate in the regulation of transducin translocation through the photoreceptor cilium. Binding of Cen2 and Cen4 to Gbetagamma of non-visual G-proteins may additionally regulate G-proteins involved in centrosome and basal body functions. PMID- 15347652 TI - Conserved transcriptional activators of the Xenopus rhodopsin gene. AB - Vertebrate rhodopsin promoters exhibit striking sequence identities proximal to the initiation site, suggesting that conserved transcription factors regulate rhodopsin expression in these animals. We identify and characterize two transcriptional activators of the Xenopus rhodopsin gene: homologs of the mammalian Crx and Nrl transcription factors, XOtx5 and XL-Nrl (originally named XL-maf), respectively. XOtx5 stimulated transcription approximately 10-fold in human 293 cells co-transfected with a plasmid containing the rhodopsin promoter ( 508 to +41) upstream of luciferase, similar to the approximately 6-fold stimulation with human Crx. XL-Nrl stimulated transcription approximately 27-fold in mammalian 293 cells co-transfected with the rhodopsin luciferase reporter, slightly more than the approximately 17-fold stimulation with Nrl. Together, the Xenopus transcription factors synergistically activated the rhodopsin promoter (approximately 140-fold), as well as in combination with mammalian homologs. Deletion of the Nrl-response element, TGCTGA, eliminated the synergistic activation by both mammalian and Xenopus transcription factors. Deletion of the conserved ATTA sequences (Ret-1 or BAT-1), binding sites for Crx, did not significantly decrease activation by Crx/XOtx5. However, there was increased activation by Nrl/XL-Nrl and an increased synergy when the Ret-1 site was disrupted. These results illustrate conservation of mechanisms of retinal gene expression among vertebrates. In transgenic tadpoles, XOtx5 and XL-Nrl directed premature and ectopic expression from the Xenopus rhodopsin promoter-GFP transgene. Furthermore, activation of the endogenous rhodopsin gene was also observed in some animals, showing that XOtx5 and XL-Nrl can activate the promoter in native chromatin environment. PMID- 15347653 TI - CCTeta, a novel soluble guanylyl cyclase-interacting protein. AB - Nitric oxide (NO) transduces most of its biological effects through activation of the heterodimeric enzyme, soluble guanylyl cyclase (sGC). Activation of sGC results in the production of cGMP from GTP. In this paper, we demonstrate a novel protein interaction between CCT (chaperonin containing t-complex polypeptide) subunit eta and the alpha1beta1 isoform of sGC. CCTeta was found to interact with the beta1 subunit of sGC via a yeast-two-hybrid screen. This interaction was then confirmed in vitro with a co-immunoprecipitation from mouse brain. The interaction between these two proteins was further supported by a co-localization of the proteins within rat brain. Using the yeast two-hybrid system, CCTeta was found to bind to the N-terminal portion of sGC. In vitro assays with purified CCTeta and Sf9 lysate expressing sGC resulted in a 30-50% inhibition of diethylamine diazeniumdiolate-NO-stimulated sGC activity. The same assays were then performed using BAY41-2272, an NO-independent allosteric sGC activator, and CCTeta had no effect on this activity. Furthermore, CCTeta had no effect on basal or sodium nitroprusside-stimulated alphabeta(Cys-105) sGC, a constitutively active mutant that only lacks the heme group. The N-terminal 94 amino acids of CCTeta seem to be critical for the mediation of this inhibition. Lastly, a 45% inhibition of sGC activity by CCTeta was seen in vivo in BE2 cells stably transfected with CCTeta and treated with sodium nitroprusside. These data suggest that CCTeta binds to sGC and, in cooperation with some other factor, inhibits its activity by modifying the binding of NO to the heme group or the subsequent conformational changes. PMID- 15347654 TI - Different mechanisms participate in the R-dependent activity of the R2R3 MYB transcription factor C1. AB - The R2R3 MYB transcription factor C1 requires the basic helix-loop-helix factor R as an essential co-activator for the transcription of maize anthocyanin genes. In contrast, the R2R3 MYB protein P1 activates a subset of the C1-regulated genes independently of R. Substitution of six amino acids in P1 with the C1 amino acids results in P1(*), whose activity on C1-regulated and P1-regulated genes is R dependent or R-enhanced, respectively. We have used P1(*) in combination with various promoters to uncover two mechanisms for R function. On synthetic promoters that contain only C1/P1 binding sites, R is an essential co-activator of C1. This function of R is unlikely to simply be the result of an increase in the C1 DNA-binding affinity, since transcriptional activity of a C1 mutant that binds DNA at a higher affinity, comparable with P1, remains R-dependent. The differential transcriptional activity of C1 fusions with the yeast Gal4 DNA binding domain in yeast and maize cells suggests that part of the function of R is to relieve C1 from a plant-specific inhibitor. A second function of R requires cis-regulatory elements in addition to the C1/P1 DNA-binding sites for R-enhanced transcription of a1. We hypothesize that R functions in this mode by binding or recruiting additional factors to the anthocyanin regulatory element conserved in the promoters of several anthocyanin genes. Together, these findings suggest a model in which combinatorial interactions with co-activators enable R2R3 MYB factors with very similar DNA binding preferences to discriminate between target genes in vivo. PMID- 15347655 TI - Human ATP:Cob(I)alamin adenosyltransferase and its interaction with methionine synthase reductase. AB - The final step in the conversion of vitamin B(12) into coenzyme B(12) (adenosylcobalamin, AdoCbl) is catalyzed by ATP:cob(I)alamin adenosyltransferase (ATR). Prior studies identified the human ATR and showed that defects in its encoding gene underlie cblB methylmalonic aciduria. Here two common polymorphic variants of the ATR that are found in normal individuals are expressed in Escherichia coli, purified, and partially characterized. The specific activities of ATR variants 239K and 239M were 220 and 190 nmol min(-1) mg(-1), and their K(m) values were 6.3 and 6.9 mum for ATP and 1.2 and 1.6 mum for cob(I)alamin, respectively. These values are similar to those obtained for previously studied bacterial ATRs indicating that both human variants have sufficient activity to mediate AdoCbl synthesis in vivo. Investigations also showed that purified recombinant human methionine synthase reductase (MSR) in combination with purified ATR can convert cob(II)alamin to AdoCbl in vitro. In this system, MSR reduced cob(II)alamin to cob(I)alamin that was adenosylated to AdoCbl by ATR. The optimal stoichiometry for this reaction was approximately 4 MSR/ATR and results indicated that MSR and ATR physically interacted in such a way that the highly reactive reaction intermediate [cob(I)alamin] was sequestered. The finding that MSR reduced cob(II)alamin to cob(I)alamin for AdoCbl synthesis (in conjunction with the prior finding that MSR reduced cob(II)alamin for the activation of methionine synthase) indicates a dual physiological role for MSR. PMID- 15347656 TI - The scaffolding A/Tpd3 subunit and high phosphatase activity are dispensable for Cdc55 function in the Saccharomyces cerevisiae spindle checkpoint and in cytokinesis. AB - Protein serine/threonine phosphatase 2A (PP2A) is a multifunctional enzyme whose trimeric form consists of a scaffolding A subunit, a catalytic C subunit, and one of several regulatory B subunits (B, B', and B''). The adenovirus E4orf4 protein associates with PP2A by directly binding the B or B' subunits. An interaction with an active PP2A containing the B subunit, or its homologue in yeast, Cdc55, is required for E4orf4-induced apoptosis in mammalian cells and for induction of growth arrest in Saccharomyces cerevisiae. In this work, Cdc55 was randomly mutagenized by low-fidelity PCR amplification, and Cdc55 mutants that lost the ability to transduce the E4orf4 toxic signal in yeast were selected. The mutations obtained by this protocol inhibited the association of Cdc55 with E4orf4, or with the PP2A-AC subunits, or both. Functional analysis revealed that a mutant that does not bind Tpd3, the yeast A subunit, as well as wild type Cdc55 in a tpd3Delta background, can form a heterodimer with the catalytic subunit. This association requires C subunit carboxyl methylation. The residual phosphatase activity associated with Cdc55 in the absence of Tpd3 is sufficient to maintain a partially active spindle checkpoint and to prevent cytokinesis defects. PMID- 15347657 TI - Cytoplasmic localization of pregnane X receptor and ligand-dependent nuclear translocation in mouse liver. AB - The pregnane X receptor (PXR) plays an important role in the response to xenobiotics and endogenous toxins. We have used a specific anti-PXR antibody in the Western blotting of mouse liver nuclear extracts to show that PXR is accumulated in the nucleus after treatment with 5-pregnen-3beta-ol-20-one-16alpha carbonitrile (PCN), followed by an increase in Cyp3a11 mRNA. Expression of wild type PXR and various mutants as green fluorescent fusion proteins in mouse livers showed that PXR was retained in the cytoplasm from where PCN treatment translocated PXR into the nucleus. Furthermore, the xenochemical response signal, the nuclear translocation signal, and the activation function 2 domain were all required for the nuclear translocation to occur. Immunoprecipitation experiments using the hsp90 antibody demonstrated the presence of PXR in a complex with the endogenous cytoplasmic constitutive active/androstane receptor retention protein (CCRP) in HepG2 cells. Fluorescence resonance energy transfer analysis of mouse liver sections after co-expression of cyan fluorescent protein-CCRP and yellow fluorescent protein-PXR also indicated that CCRP and PXR were closely associated in vivo. Overexpression of exogenous CCRP increased the cytoplasmic level of the PXR.CCRP.hsp90 complex, whereas a decrease in endogenous CCRP by treatment with small interfering RNA for CCRP repressed the PXR-mediated reporter activity in HepG2 cells. We conclude that the CCRP mediates the retention of PXR in the cytosol and modulates the activation of PXR in response to PCN treatment. PMID- 15347658 TI - Catalase reaction by myoglobin mutants and native catalase: mechanistic investigation by kinetic isotope effect. AB - The catalase reaction has been studied in detail by using myoglobin (Mb) mutants. Compound I of Mb mutants (Mb-I), a ferryl species (Fe(IV)=O) paired with a porphyrin radical cation, is readily prepared by the reaction with a nearly stoichiometric amount of m-chloroperbenzoic acid. Upon the addition of H2O2 to an Mb-I solution, Mb-I is reduced back to the ferric state without forming any intermediates. This indicates that Mb-I is capable of performing two-electron oxidation of H2O2 (catalatic reaction). Gas chromatography-mass spectroscopy analysis of the evolved O2 from a 50:50 mixture of H2(18)O2/H2(16)O2 solution containing H64D or F43H/H64L Mb showed the formation of 18O2 (m/e = 36) and 16O2 (m/e = 32) but not 16O18O (m/e = 34). This implies that O2 is formed by two electron oxidation of H2O2 without breaking the O-O bond. Deuterium isotope effects on the catalatic reactions of Mb mutants and catalase suggest that the catalatic reactions of Micrococcus lysodeikticus catalase and F43H/H64L Mb proceed via an ionic mechanism with a small isotope effect of less than 4.0, since the distal histidine residue is located at a proper position to act as a general acid-base catalyst for the ionic reaction. In contrast, other Mb mutants such as H64X (X is Ala, Ser, and Asp) and L29H/H64L Mb oxidize H2O2 via a radical mechanism in which a hydrogen atom is abstracted by Mb-I with a large isotope effect in a range of 10-29, due to a lack of the general acid-base catalyst. PMID- 15347659 TI - DNA replication checkpoint control mediated by the spindle checkpoint protein Mad2p in fission yeast. AB - The relationship between the DNA replication and spindle checkpoints of the cell cycle is unclear, given that in most eukaryotes, spindle formation occurs only after DNA replication is complete. Fission yeast rad3 mutant cells, which are deficient in DNA replication checkpoint function, enter, progress through, and exit mitosis even when DNA replication is blocked. In contrast, the entry of cds1 mutant cells into mitosis is delayed by several hours when DNA replication is inhibited. We show here that this delay in mitotic entry in cds1 cells is due in part to activation of the spindle checkpoint protein Mad2p. In the presence of the DNA replication inhibitor hydroxyurea (HU), cds1 mad2 cells entered and progressed through mitosis earlier than did cds1 cells. Overexpression of Mad2p or inactivation of Slp1p, a regulator of the anaphase-promoting complex, also rescued the checkpoint defect of HU-treated rad3 cells. Rad3p was shown to be involved in the physical interaction between Mad2p and Slp1p in the presence of HU. These results suggested that Mad2p and Slp1p act downstream of Rad3p in the DNA replication checkpoint and that Mad2p is required for the DNA replication checkpoint when Cds1p is compromised. PMID- 15347660 TI - The critical role of the 185-189-loop in the factor Xa interaction with Na+ and factor Va in the prothrombinase complex. AB - The S1 site (Asp(189)) of factor Xa (fXa) is located on a loop (residues 185-189) that contains three solvent-exposed charged residues (Asp(185), Lys(186), and Glu(188)) below the active-site pocket of the protease. To investigate the role of these residues in the catalytic function of fXa, we expressed three mutants of the protease in which the charges of these residues were neutralized by their substitutions with Ala (D185A, K186A, and E188A). Kinetic studies revealed that E188A has a normal catalytic activity toward small synthetic and natural substrates and inhibitors of fXa; however, the same activities were slightly ( approximately 2-fold) and dramatically ( approximately 20-50-fold) impaired for the D185A and K186A mutants, respectively. Further studies revealed that the affinity of D185A and K186A for interaction with Na(+) has also been altered, with a modest impairment ( approximately 2-fold) for the former and a dramatic impairment for the latter mutant. Both prothrombinase and direct binding studies indicated that K186A also has an approximately 6-fold impaired affinity for factor Va. Interestingly, a saturating concentration of factor Va restored the catalytic defect of K186A in reactions with prothrombin and the recombinant tick anticoagulant peptide that is known to interact with the Na(+) loop of fXa, but not with other substrates. These results suggest that factor Va interacts with 185-189-loop for fXa, which is energetically linked to the Na(+)-binding site of the protease. PMID- 15347661 TI - cAMP-dependent protein kinase regulates ubiquitin-proteasome-mediated degradation and subcellular localization of the nuclear receptor coactivator GRIP1. AB - Nuclear receptors and their coactivators are key regulators of numerous physiological functions. GRIP1 (glucocorticoid receptor-interacting protein) is a member of the steroid receptor coactivator family. Here, we show that GRIP1 is regulated by cAMP-dependent protein kinase (PKA) that induces its degradation through the ubiquitin-proteasome pathway. GRIP1 was down-regulated in transiently transfected COS-1 cells after treatment with 8-para-chlorophenylthio-cAMP or forskolin and 3-isobutyl-1-methylxanthine and in adrenocortical Y1 cells after incubation with adrenocorticotropic hormone. Pulse-chase experiments with transiently transfected COS-1 cells demonstrated that the half-life of GRIP1 was markedly reduced in cells overexpressing the PKA catalytic subunit, suggesting that activation of PKA increases the turnover of GRIP1 protein. The proteasome inhibitors MG132 and lactacystin abolished the PKA-mediated degradation of GRIP1. Using ts20 cells, a temperature-sensitive cell line that contains a thermolabile ubiquitin-activating E1 enzyme, it was confirmed that PKA-mediated degradation of GRIP1 is dependent upon the ubiquitin-proteasome pathway. Coimmunoprecipitation studies of COS-1 cells transfected with expression vectors encoding GRIP1 and ubiquitin using anti-GRIP1 and anti-ubiquitin antibodies showed that the ubiquitination of GRIP1 was increased by overexpression of PKA. Finally, we show that PKA regulates the intracellular distribution pattern of green fluorescent protein-GRIP1 and stimulates recruitment of GRIP1 to subnuclear foci that are colocalized with the proteasome. Taken together, these data demonstrate that GRIP1 is ubiquitinated and degraded through activation of the PKA pathway. This may represent a novel regulatory mechanism whereby hormones down-regulate a nuclear receptor coactivator. PMID- 15347662 TI - ATP-binding cassette transporter A1 contains a novel C-terminal VFVNFA motif that is required for its cholesterol efflux and ApoA-I binding activities. AB - The stimulation of cellular cholesterol and phospholipid efflux by apolipoprotein A-I is mediated by the activity of the ATP-binding cassette transporter A1 (ABCA1). Individuals with Tangier disease harbor loss-of-function mutations in this transporter that have proven useful in illuminating its activity. Here, we analyze a mutation that deletes the last 46 residues of the 2261 amino acid transporter (Delta46) and eliminates its lipid efflux. As the final four amino acids of the C terminus represent a putative PDZ-binding motif, we initially characterized deletion mutants lacking only these residues. Although a moderate decline in lipid efflux was detected, this decline was not as profound as that seen in the Delta46 mutant. Subsequent systematic analysis of the ABCA1 C terminus revealed a novel, highly conserved motif (VFVNFA) that was required for lipid efflux. Alteration of this motif, which is present in some but not all members of the ABCA family, did not prevent trafficking of the transporter to the plasma membrane but did eliminate its binding of apoA-I. Chimeric transporters, generated by substituting the C termini of either ABCA4 or ABCA7 for the endogenous terminus, demonstrated that ABCA1 could stimulate cholesterol efflux without its PDZ-binding motif but not without the VFVNFA motif. When a peptide containing the VFVNFA sequence was introduced into ABCA1-expressing cells, ABCA1 mediated lipid efflux was also markedly inhibited. These results indicate that the C-terminal VFVNFA motif of ABCA1 is essential for its lipid efflux activity. The data also suggest that this motif participates in novel protein-protein interactions that may be shared among members of the ABCA family. PMID- 15347663 TI - Osmotic response element-binding protein (OREBP) is an essential regulator of the urine concentrating mechanism. AB - OREBP (osmotic response element-binding protein), also called TonEBP or NFAT5, is thought to induce the expression of genes that increase the accumulation of organic osmolytes to protect cells against a hypertonic environment. To investigate the consequences of lacking OREBP activity, transgenic (Tg) mice that overexpress OREBPdn (dominant negative form of OREBP) specifically in the epithelial cells of the renal collecting tubules were generated. These mice showed impairment in their urine concentrating mechanism, most likely due to reduced expression of the aquaporin AQP2 and the urea transporter UT-A1 and UT-A2 mRNAs. When deprived of water or after the administration of a vasopressin analogue, urine osmolality of the Tg mice was significantly increased but not to the same extent as that of the wild type mice. The expression of AQP2 and UT-A1, but not UT-A2 mRNAs, was increased to the same level as that of the wild type mice in the water deprivation state, indicating that the vasopressin regulatory mechanism was not affected by OREBPdn. These data indicate that in addition to vasopressin, OREBP is another essential regulator of the urine concentrating mechanism. Furthermore, the OREBPdn Tg mice developed progressive hydronephrosis soon after weaning, confirming the osmoprotective function of OREBP implicated by the in vitro experiments. PMID- 15347664 TI - Targeting of NPC1 to late endosomes involves multiple signals, including one residing within the putative sterol-sensing domain. AB - The NPC1 protein is a multipass transmembrane protein whose deficiency causes the autosomal recessive lipid storage disorder Niemann-Pick type C1. NPC1 localizes predominantly to late endosomes and has a dileucine motif located within a small cytoplasmic tail thought to target the protein to this location. Our data have suggested previously that the protein can reach its correct location in the absence of its cytoplasmic tail, suggesting that other signals contribute to NPC1 targeting. By using various FLAG-tagged and CD32-NPC1 chimeric fusion constructs, we show that multiple signals are responsible for the trafficking of NPC1 to the endosomal compartment, including the dileucine motif and a previously unidentified signal residing within the putative sterol-sensing domain transmembrane domain 3. Neither region alone was capable of directing heterologous CD32 fusions to late endosomes exclusively via the trans-Golgi network to the late endosome route taken by wild-type NPC1; transmembrane domain 3 was unable to maintain CD32 in late endosomes, indicating that two or more signals work in concert to target and retain NPC1 in this compartment. In addition we confirm that the tail dileucine motif is not essential for NPC1 targeting to late endosomes, and we discuss the implications of this finding along with the previously unappreciated role for transmembrane domain 3 in NPC1 localization and function. PMID- 15347665 TI - Rab5-associated vacuoles play a unique role in phagocytosis of the enteric protozoan parasite Entamoeba histolytica. AB - In mammals, Rab5 and Rab7 play a specific and coordinated role in a sequential process during phagosome maturation. Here, we report that Rab5 and Rab7 in the enteric protozoan parasite Entamoeba histolytica, EhRab5 and EhRab7A, are involved in steps that are distinct from those known for mammals. EhRab5 and EhRab7A were localized to independent small vesicular structures at steady state. Priming with red blood cells induced the formation of large vacuoles associated with both EhRab5 and EhRab7A ("prephagosomal vacuoles (PPV)") in the amoeba within an incubation period of 5-10 min. PPV emerged de novo physically and distinct from phagosomes. PPV were gradually acidified and matured by fusion with lysosomes containing a digestive hydrolase, cysteine proteinase, and a membrane permeabilizing peptide amoebapore. After EhRab5 dissociated from PPV, 5-10 min later, the EhRab7A-PPV fused with phagosomes, and EhRab7A finally dissociated from the phagosomes. Immunoelectron and light micrographs showed that PPV contained small vesicle-like structures containing fluid-phase markers and amoebapores, which were not evenly distributed within PPV, suggesting that the mechanism was similar to multivesicular body formation in PPV generation. In contrast to Rab5 from other organisms, EhRab5 was involved exclusively in phagocytosis, but not in endocytosis. Overexpression of wild-type EhRab5 enhanced phagocytosis and the transport of amoebapore to phagosomes. Conversely, expression of an EhRab5Q67L GTP form mutant impaired the formation of PPV and phagocytosis. Altogether, we propose that the amoebic Rab5 plays an important role in the formation of unique vacuoles, which is essential for engulfment of erythrocytes and important for packaging of lysosomal hydrolases, prior to the targeting to phagosomes. PMID- 15347666 TI - Blockade of electron transport during ischemia protects cardiac mitochondria. AB - Subsarcolemmal mitochondria sustain progressive damage during myocardial ischemia. Ischemia decreases the content of the mitochondrial phospholipid cardiolipin accompanied by a decrease in cytochrome c content and a diminished rate of oxidation through cytochrome oxidase. We propose that during ischemia mitochondria produce reactive oxygen species at sites in the electron transport chain proximal to cytochrome oxidase that contribute to the ischemic damage. Isolated, perfused rabbit hearts were treated with rotenone, an irreversible inhibitor of complex I in the proximal electron transport chain, immediately before ischemia. Rotenone pretreatment preserved the contents of cardiolipin and cytochrome c measured after 45 min of ischemia. The rate of oxidation through cytochrome oxidase also was improved in rotenone-treated hearts. Inhibition of the electron transport chain during ischemia lessens damage to mitochondria. Rotenone treatment of isolated subsarcolemmal mitochondria decreased the production of reactive oxygen species during the oxidation of complex I substrates. Thus, the limitation of electron flow during ischemia preserves cardiolipin content, cytochrome c content, and the rate of oxidation through cytochrome oxidase. The mitochondrial electron transport chain contributes to ischemic mitochondrial damage that in turn augments myocyte injury during subsequent reperfusion. PMID- 15347667 TI - Physiological roles of the intermediate conductance, Ca2+-activated potassium channel Kcnn4. AB - Three broad classes of Ca(2+)-activated potassium channels are defined by their respective single channel conductances, i.e. the small, intermediate, and large conductance channels, often termed the SK, IK, and BK channels, respectively. SK channels are likely encoded by three genes, Kcnn1-3, whereas IK and most BK channels are most likely products of the Kcnn4 and Slo (Kcnma1) genes, respectively. IK channels are prominently expressed in cells of the hematopoietic system and in organs involved in salt and fluid transport, including the colon, lung, and salivary glands. IK channels likely underlie the K(+) permeability in red blood cells that is associated with water loss, which is a contributing factor in the pathophysiology of sickle cell disease. IK channels are also involved in the activation of T lymphocytes. The fluid-secreting acinar cells of the parotid gland express both IK and BK channels, raising questions about their particular respective roles. To test the physiological roles of channels encoded by the Kcnn4 gene, we constructed a mouse deficient in its expression. Kcnn4 null mice were of normal appearance and fertility, their parotid acinar cells expressed no IK channels, and their red blood cells lost K(+) permeability. The volume regulation of T lymphocytes and erythrocytes was severely impaired in Kcnn4 null mice but was normal in parotid acinar cells. Despite the loss of IK channels, activated fluid secretion from parotid glands was normal. These results confirm that IK channels in red blood cells, T lymphocytes, and parotid acinar cells are indeed encoded by the Kcnn4 gene. The role of these channels in water movement and the subsequent volume changes in red blood cells and T lymphocytes is also confirmed. Surprisingly, Kcnn4 channels appear to play no required role in fluid secretion and regulatory volume decrease in the parotid gland. PMID- 15347668 TI - Modeling a sialic acid binding pocket in the external loops of JC virus VP1. AB - JC virus (JCV) is a common human polyomavirus that infects over 70% of the population worldwide. JCV has a restricted cell tropism that is caused partly by the initial interaction between the virus and sialic acid-containing host cell receptors. To identify the molecular interactions between the virus and its cellular receptor, we used a combined approach of site-directed mutagenesis and homology-based molecular modeling. A model of the major viral capsid protein VP1 based on sequence alignment with other closely related polyomaviruses allowed us to target specific amino acids in the extracellular loops of VP1 for mutagenesis. An analysis of the growth rates of 17 point mutants led to the identification of VP1 amino acids that are critical in virus-host cell receptor interactions. Molecular dynamics simulations were then used to build and confirm a model of the interaction between VP1 and the sialic acid component of the JCV receptor. PMID- 15347669 TI - Roles of Brahma and Brahma/SWI2-related gene 1 in hypoxic induction of the erythropoietin gene. AB - Upon hypoxia, the human erythropoietin (EPO) gene is transactivated by the heterodimeric hypoxia-inducible factor 1 (HIF-1). Mammalian SWI/SNF is a chromatin-remodeling complex involved in the modulation of gene expression. We demonstrate that Brahma (Brm) and Brahma/SWI2-related gene 1 (Brg-1), alternative ATPase subunits of SWI/SNF, potentiate reporter gene activation mediated by HIF-1 in an ATPase-dependent manner. Brm was more potent than Brg-1 in the reporter gene assays. Simultaneous depletion of both Brm and Brg-1 by small interfering RNAs significantly compromised the transcription of the endogenous EPO gene triggered by hypoxia. Whereas knocking down Brm alone resulted in a moderate reduction in transcription of the EPO gene, depletion of Brg-1 resulted in an augmentation of transcription of both the EPO gene and the Brm gene, indicating that Brm can compensate for loss of Brg-1. Chromatin immunoprecipitation (ChIP) and sequential ChIP (re-ChIP) analysis showed that both Brm and Brg-1 associate with the enhancer region of the EPO gene in vivo in a hypoxia-dependent fashion and that each is present in a complex with HIF-1. Brm and Brg-1 were also recruited to the promoter of the vascular endothelial growth factor (VEGF) gene in a hypoxia-dependent fashion, although hypoxic induction of VEGF transcription was not affected by depletions of either or both Brm and Brg-1. Together these studies reveal a novel role for SWI/SNF in the activation of transcription of the EPO gene, indicate an important communication and compensation between Brm and Brg-1, and suggest that the requirement for SWI/SNF during hypoxic induction is gene-specific. PMID- 15347670 TI - Cystathionine gamma-lyase overexpression inhibits cell proliferation via a H2S dependent modulation of ERK1/2 phosphorylation and p21Cip/WAK-1. AB - Cystathionine gamma-lyase (CSE) is a key enzyme in the trans-sulfuration pathway. CSE uses L-cysteine as a substrate to produce hydrogen sulfide (H2S). The CSE/H2S system has been shown to play an important role in regulating cellular functions in different systems. In the present study, we used CSE stably overexpressed HEK 293 cells to explore the effect of the CSE/H2S system on cell growth and proliferation. The overexpression of CSE resulted in increases in CSE mRNA levels, CSE proteins, and intracellular H2S production rates, as well as the inhibition of cell proliferation and DNA synthesis. These effects were accompanied by a sustained ERK activation and up-regulation of the cyclin dependent kinase inhibitor p21Cip/WAK-1. Blocking the action of ERK with U0126 inhibited the induction of p21Cip/WAK-1, suggesting that ERK activation functions upstream of p21Cip/WAK-1 activation to initiate the CSE overexpression-induced cell growth inhibition. The antiproliferative effect of CSE is likely mediated by endogenously produced H2S because the H2S scavenger methemoglobin (10 microm) significantly decreased the H2S production rate and reversed the antiproliferative effect afforded by CSE. Exogenous H2S (100 microm) also inhibited cell proliferation. However, the other CSE-catalyzed products, ammonium and pyruvate, failed to inhibit cell proliferation. Methemoglobin also abolished the inhibitory effect of exogenous H2S on cell proliferation. Moreover, exogenous H2S induced a sustained ERK and p21Cip/WAK-1 activation. These findings support the hypothesis that endogenously produced H2S may play a fundamental role in cell proliferation and survival. PMID- 15347671 TI - Sequential 2'-O-methylation of archaeal pre-tRNATrp nucleotides is guided by the intron-encoded but trans-acting box C/D ribonucleoprotein of pre-tRNA. AB - Haloferax volcanii pre-tRNA(Trp) processing requires box C/D ribonucleoprotein (RNP)-guided 2'-O-methylation of nucleotides C34 and U39 followed by intron excision. Positioning of the box C/D guide RNA within the intron of this pre-tRNA led to the assumption that nucleotide methylation is guided by the cis-positioned box C/D RNPs. We have now investigated the mechanism of 2'-O-methylation for the H. volcanii pre-tRNA(Trp) in vitro by assembling methylation-competent box C/D RNPs on both the pre-tRNA and the excised intron (both linear and circular forms) using Methanocaldococcus jannaschii box C/D RNP core proteins. With both kinetic studies and single nucleotide substitutions of target and guide nucleotides, we now demonstrate that pre-tRNA methylation is guided in trans by the intron encoded box C/D RNPs positioned in either another pre-tRNA(Trp) or in the excised intron. Methylation by in vitro assembled RNPs prefers but does not absolutely require Watson-Crick pairing between the guide and target nucleotides. We also demonstrate for the first time that methylation of two nucleotides guided by a single box C/D RNA is sequential, that is, box C'/D' RNP-guided U39 methylation first requires box C/D RNP-guided methylation of C34. Methylation of the two nucleotides of exogenous pre-tRNA(Trp) added to an H. volcanii cell extract also occurs sequentially and is also accomplished in trans using RNPs that pre-exist in the extract. Thus, this trans mechanism is analogous to eukaryal pre-rRNA 2'-O methylation guided by intron-encoded but trans-acting box C/D small nucleolar RNPs. This trans mechanism could explain the observed accumulation of the excised H. volcanii pre-tRNA(Trp) intron in vivo. A trans mechanism would also eliminate the obligatory refolding of the pre-tRNA that would be required to carry out two cis-methylation reactions before pre-tRNA splicing. PMID- 15347672 TI - Membrane-embedded C-terminal segment of rat mitochondrial TOM40 constitutes protein-conducting pore with enriched beta-structure. AB - TOM40 is the central component of the preprotein translocase of the mitochondrial outer membrane (TOM complex). We purified recombinant rat TOM40 (rTOM40), which was refolded in Brij35 after solubilization from inclusion bodies by guanidine HCl. rTOM40 (i) consisted of a 63% beta-sheet structure and (ii) bound a matrix targeted preprotein with high affinity and partially translocated it into the rTOM40 pore. This partial translocation was inhibited by stabilization of the mature domain of the precursor. (iii) rTOM40 bound preprotein initially through ionic interactions, followed by salt-resistant non-ionic interactions, and (iv) exhibited presequence-sensitive, cation-specific channel activity in reconstituted liposomes. Based on the domain structure of rTOM40 deduced by protease treatment, we purified the elastase-resistant and membrane-embedded C terminal segment (rTOM40(DeltaN165)) as a recombinant protein with 62% beta structure that exhibited properties comparable with those of full-size rTOM40. We concluded that the membrane-embedded C-terminal half of rTOM40 constitutes the preprotein recognition domain with an enriched beta-structure, which forms the preprotein conducting pore containing a salt-sensitive cis-binding site and a salt-resistant trans-binding site. PMID- 15347673 TI - Endothelin-1-induced prostaglandin E2-EP2, EP4 signaling regulates vascular endothelial growth factor production and ovarian carcinoma cell invasion. AB - Cyclooxygenase (COX)-1- and COX-2-derived prostaglandins are implicated in the development and progression of several malignancies. We have recently demonstrated that treatment of ovarian carcinoma cells with endothelin-1 (ET-1) induces expression of both COX-1 and COX-2, which contributes to vascular endothelial growth factor (VEGF) production. In this study, we show that in HEY and OVCA 433 ovarian carcinoma cells, ET-1, through the binding with ETA receptor (ETAR), induces prostaglandin E2 (PGE2) production, as the more represented PG types, and increases the expression of PGE2 receptor type 2 (EP2) and type 4 (EP4). The use of pharmacological EP agonists and antagonists indicates that ET-1 and PGE2 stimulate VEGF production principally through EP2 and EP4 receptors. At the mechanistic level, we prove that the induction of PGE2 and VEGF by ET-1 involves Src-mediated epidermal growth factor receptor transactivation. Finally, we demonstrate that ETAR-mediated activation of PGE2-dependent signaling participates in the regulation of the invasive behavior of ovarian carcinoma cells by activating tumor-associated matrix metalloproteinase. These results implicate EP2 and EP4 receptors in the induction of VEGF expression and cell invasiveness by ET-1 and provide a mechanism by which ETAR/ET-1 can promote and interact with PGE2-dependent machinery to amplify its proangiogenic and invasive phenotype in ovarian carcinoma cells. Pharmacological blockade of ETAR can therefore represent an additional strategy to control PGE2 signaling, which has been associated with ovarian carcinoma progression. PMID- 15347674 TI - Role of the tetradecapeptide repeat domain of human histone deacetylase 6 in cytoplasmic retention. AB - Histone deacetylase 6 (HDAC6) contains tandem catalytic domains and a ubiquitin binding zinc finger and displays deacetylase activity toward acetylated microtubules. Here we show that unlike its orthologs from Caenorhabditis elegans, Drosophila, and mouse, human HDAC6 possesses a tetradecapeptide repeat domain located between the second deacetylase domain and the C-terminal ubiquitin binding motif. Related to this structural difference, the cytoplasmic localization of human, but not murine, HDAC6 is resistant to treatment with leptomycin B (LMB). Although it is dispensable for the deacetylase and ubiquitin binding activities of human HDAC6, the tetradecapeptide repeat domain displays acetyl-microtubule targeting ability. Moreover, it forms a unique structure and is required for the LMB-resistant cytoplasmic localization of human HDAC6. Besides the tetradecapeptide repeat domain, human HDAC6 possesses two LMB sensitive nuclear export signals and a nuclear localization signal. These results thus indicate that the cytoplasmic localization for murine and human HDAC6 proteins is differentially regulated and suggest that the tetradecapeptide repeat domain serves as an important sequence element to stably retain human HDAC6 in the cytoplasm. PMID- 15347675 TI - Protein kinase C mediates phosphorylation, desensitization, and trafficking of the D2 dopamine receptor. AB - Previously, D2 dopamine receptors (D2 DARs) have been shown to undergo G-protein coupled receptor kinase phosphorylation in an agonist-specific fashion. We have now investigated the ability of the second messenger-activated protein kinases, protein kinase A (PKA) and protein kinase C (PKC), to mediate phosphorylation and desensitization of the D2 DAR. HEK293T cells were transiently transfected with the D2 DAR and then treated with intracellular activators and inhibitors of PKA or PKC. Treatment with agents that increase cAMP, and activate PKA, had no effect on the phosphorylation state of the D2 DAR, suggesting that PKA does not phosphorylate the D2 DAR in HEK293T cells. In contrast, cellular treatment with phorbol 12-myristate 13-acetate (PMA), a PKC activator, resulted in an approximately 3-fold increase in D2 DAR phosphorylation. The phosphorylation was specific for PKC as the PMA effect was mimicked by phorbol 12,13-dibutyrate, but not by 4alpha-phorbol 12,13-didecanoate, active and inactive, phorbol diesters, respectively. The PMA-mediated D2 DAR phosphorylation was completely blocked by co-treatment with the PKC inhibitor, bisindolylmaleimide II, and augmented by co transfection with PKCbetaI. In contrast, PKC inhibition had no effect on agonist promoted phosphorylation, suggesting that PKC is not involved in this response. PKC phosphorylation of the D2 DAR was found to promote receptor desensitization as reflected by a decrease in agonist potency for inhibiting cAMP accumulation. Most interestingly, PKC phosphorylation also promoted internalization of the D2 DAR through a beta-arrestin- and dynamin-dependent pathway, a response not usually associated with PKC phosphorylation of G-protein-coupled receptors. Site directed mutagenesis experiments resulted in the identification of two domains of PKC phosphorylation sites within the third intracellular loop of the receptor. Both of these domains are involved in regulating sequestration of the D2 DAR, whereas only one domain is involved in receptor desensitization. These results indicate that PKC can mediate phosphorylation of the D2 DAR, resulting in both functional desensitization and receptor internalization. PMID- 15347676 TI - Sites of the NUDT9-H domain critical for ADP-ribose activation of the cation channel TRPM2. AB - TRPM2 is a cation channel unique within the transient receptor potential family because of its gating by ADP-ribose (ADPR). ADPR gating is enabled by a cytosolic C-terminal Nudix box sequence motif embedded into a region homologous to the NUDT9 ADPR pyrophosphatase. A recently discovered splice variant of TRPM2 (TRPM2 DeltaC) lacks 34 amino acid residues in the NUDT9 domain and is insensitive to ADPR. To analyze in detail which parts of the deleted sequence (DeltaC-stretch) are critical for ADPR gating, we tested mutants that lacked 19, 25, and 29 amino acid residues in the N-terminal part or had amino acid residues substituted in the remaining C-terminal part of the DeltaC-stretch. All of these mutants displayed typical ADPR-induced currents. However, the deletion or substitution of the amino acid residue Asn-1326 immediately downstream of the DeltaC-stretch abrogated ADPR gating. We furthermore analyzed the mutation I1405E/L1406F in the Nudix box of TRPM2, because a considerably decreased AD-PRase activity of the TRPM2 NUDT9-H protein in comparison to the NUDT9 pyrophosphatase has been attributed to the reverse exchange EF --> IL. The I1405E/L1406F variant of TRPM2 failed to respond to ADPR even at concentrations up to 10 mM. We concluded that the DeltaC-stretch contains no individual amino acid residues essential for ADPR gating but may act as a spacer segment stabilizing a conformation necessary for the essential residue Asn-1326 to interact with other channel regions. Enhancing the enzymatic activity of the Nudix box abolishes the ADPR gating of TRPM2, pointing to the requirement of prolonged binding rather than degradation. PMID- 15347677 TI - Impaired tricarboxylic acid cycle activity in mouse livers lacking cytosolic phosphoenolpyruvate carboxykinase. AB - Liver-specific phosphoenolpyruvate carboxykinase (PEPCK) null mice, when fasted, maintain normal whole body glucose kinetics but develop dramatic hepatic steatosis. To identify the abnormalities of hepatic energy generation that lead to steatosis during fasting, we studied metabolic fluxes in livers lacking hepatic cytosolic PEPCK by NMR using 2H and 13C tracers. After a 4-h fast, glucose production from glycogenolysis and conversion of glycerol to glucose remains normal, whereas gluconeogenesis from tricarboxylic acid (TCA) cycle intermediates was nearly absent. Upon an extended 24-h fast, livers that lack PEPCK exhibit both 2-fold lower glucose production and oxygen consumption, compared with the controls, with all glucose production being derived only from glycerol. The mitochondrial reduction-oxidation (red-ox) state, as indicated by the NADH/NAD+ ratio, is 5-fold higher, and hepatic TCA cycle intermediate concentrations are dramatically increased in the PEPCK null livers. Consistent with this, flux through the TCA cycle and pyruvate cycling pathways is 10- and 40 fold lower, respectively. Disruption of hepatic cataplerosis due to loss of PEPCK leads to the accumulation of TCA cycle intermediates and a nearly complete blockage of gluconeogenesis from amino acids and lactate (an energy demanding process) but intact gluconeogenesis from glycerol (which contributes to net NADH production). Inhibition of the TCA cycle and fatty acid oxidation due to increased TCA cycle intermediate concentrations and reduced mitochondrial red-ox state lead to the development of steatosis. PMID- 15347679 TI - Accumulation of the D2 protein is a key regulatory step for assembly of the photosystem II reaction center complex in Synechocystis PCC 6803. AB - Accumulation of monomer and dimer photosystem (PS) II reaction center core complexes has been analyzed by two-dimensional Blue-native/SDS-PAGE in Synechocystis PCC 6803 wild type and in mutant strains lacking genes psbA, psbB, psbC, psbDIC/DII, or the psbEFLJ operon. In vivo pulse-chase radiolabeling experiments revealed that mutant cells assembled PSII precomplexes only. In DeltapsbC and DeltapsbB, assembly of reaction center cores lacking CP43 and reaction center complexes was detected, respectively. In DeltapsbA, protein subunits CP43, CP47, D2, and cytochrome b559 were synthesized, but proteins did not assemble. Similarly, in DeltapsbD/C lacking D2, and CP43, the de novo synthesized proteins D1, CP47, and cytochrome b559 did not form any mutual complexes, indicating that assembly of the reaction center complex is a prerequisite for assembly with core subunits CP47 and CP43. Finally, although CP43 and CP47 accumulated in DeltapsbEFLJ, D2 was neither expressed nor accumulated. We, furthermore, show that the amount of D2 is high in the strain lacking D1, whereas the amount of D1 is low in the strain lacking D2. We conclude that expression of the psbEFLJ operon is a prerequisite for D2 accumulation that is the key regulatory step for D1 accumulation and consecutive assembly of the PSII reaction center complex. PMID- 15347678 TI - Interleukin (IL)-15 and IL-2 reciprocally regulate expression of the chemokine receptor CX3CR1 through selective NFAT1- and NFAT2-dependent mechanisms. AB - We have recently reported that interleukin (IL)-15 and IL-2, which signal through IL-2Rbetagamma, oppositely regulate expression of the proinflammatory chemokine receptor CX3CR1. Here we delineate molecular mechanisms responsible for this paradox. By using a luciferase reporter plasmid, we identified a 433-bp region spanning the major transcriptional start point of human CX3CR1 that, when expressed in human peripheral blood mononuclear cells (PBMCs), possessed strong constitutive promoter activity. IL-2 and IL-15 treatment increased and abolished this activity, respectively, mimicking their effects on endogenous CX3CR1. IL-2 and IL-15 have been reported to also have opposite effects on the immunoregulatory transcription factor NFAT (nuclear factor of activated T cells), and the 433-bp region contains a kappaB-like NFAT site. The effects of IL-15 and IL-2 on both CX3CR1 reporter activity and endogenous CX3CR1 transcription in PBMCs were abolished by the NFAT inhibitors cyclosporin A and VIVIT. Moreover, mutation of the kappaB-like NFAT sequence markedly attenuated IL-2 and IL-15 modulation of CX3CR1 promoter-reporter activity in PBMCs. Furthermore, chromatin immunoprecipitation revealed that IL-15 promoted specific recruitment of NFAT1 but not NFAT2 to the CX3CR1 promoter, whereas IL-2 had the converse effect. This appears to be relevant in vivo because mouse CX3CR1 mRNA was expressed in both PBMCs and splenocytes from NFAT1-/- mice injected with recombinant IL-15 but was undetectable in cells from IL-15-injected NFAT1+/+ BALB/c mice; as predicted, IL 2 up-regulated cx3cr1 in both mouse strains to a similar extent. Thus, by pharmacologic, genetic, and biochemical criteria in vitro and in vivo, our results suggest that IL-15 and IL-2 oppositely regulate CX3CR1 gene expression by differentially recruiting NFAT1 and NFAT2 to a kappaB-like NFAT site within the CX3CR1 promoter. We propose that expression of CX3CR1 and possibly other immunoregulatory genes may be determined in part by the balance of NFAT1 and NFAT2 activity in leukocytes. PMID- 15347680 TI - Inactivation of CaMIT1 inhibits Candida albicans phospholipomannan beta mannosylation, reduces virulence, and alters cell wall protein beta mannosylation. AB - Studies on Candida albicans phospholipomannan have suggested a novel biosynthetic pathway for yeast glycosphingolipids. This pathway is thought to diverge from the usual pathway at the mannose-inositol-phospho-ceramide (MIPC) step. To confirm this hypothesis, a C. albicans gene homologue for the Saccharomyces cerevisiae SUR1 gene was identified and named MIT1 as it coded for GDP-mannose:inositol phospho-ceramide mannose transferase. Two copies of this gene were disrupted. Western blots of cell extracts revealed that strain mit1Delta contained no PLM. Thin layer chromatography and mass spectrometry confirmed that mit1Delta did not synthesize MIPC, demonstrating a role of MIT1 in the mannosylation of C. albicans IPCs. As MIT1 disruption prevented downstream beta-1,2 mannosylation, mit1Delta represents a new C. albicans mutant affected in the expression of these specific virulence attributes, which act as adhesins/immunomodulators. mit1Delta was less virulent during both the acute and chronic phases of systemic infection in mice (75 and 50% reduction in mortality, respectively). In vitro, mit1Delta was not able to escape macrophage lysis through down-regulation of the ERK1/2 phosphorylation pathway previously shown to be triggered by PLM. Phenotypic analysis also revealed pleiotropic effects of MIT1 disruption. The most striking observation was a reduced beta-mannosylation of phosphopeptidomannan. Increased beta-mannosylation of mannoproteins was observed under growth conditions that prevented the association of beta-oligomannosides with phosphopeptidomannan, but not with PLM. This suggests that C. albicans has strong regulatory mechanisms associating beta-oligomannoses with different cell wall carrier molecules. These mechanisms and the impact of the different presentations of beta-oligomannoses on the host response need to be defined. PMID- 15347681 TI - Translational regulation of Hsp90 mRNA. AUG-proximal 5'-untranslated region elements essential for preferential heat shock translation. AB - Heat shock in Drosophila results in repression of most normal (non-heat shock) mRNA translation and the preferential translation of the heat shock mRNAs. The sequence elements that confer preferential translation have been localized to the 5'-untranslated region (5'-UTR) for Hsp22 and Hsp70 mRNAs (in Drosophila). Hsp90 mRNA is unique among the heat shock mRNAs in having extensive secondary structure in its 5'-UTR and being abundantly represented in the non-heat shocked cell. In this study, we show that Hsp90 mRNA translation is inefficient at normal growth temperature, and substantially activated by heat shock. Its preferential translation is not based on an IRES-mediated translation pathway, because overexpression of eIF4E-BP inhibits its translation (and the translation of Hsp70 mRNA). The ability of Hsp90 mRNA to be preferentially translated is conferred by its 5'-UTR, but, in contrast to Hsp22 and -70, is primarily influenced by nucleotides close to the AUG initiation codon. We present a model to account for Hsp90 mRNA translation, incorporating results indicating that heat shock inhibits eIF4F activity, and that Hsp90 mRNA translation is sensitive to eIF4F inactivation. PMID- 15347682 TI - Characterization of a novel interaction between the secretory Na+-K+-Cl- cotransporter and the chaperone hsp90. AB - The first isoform of the Na(+)-K(+)-Cl(-) cotransporter (NKCC1) is of central importance for the control of cellular ion concentration and epithelium-mediated salt secretion. Several studies have established that a change in intracellular [Cl(-)] (Cl(-)(i)) represents a key signaling mechanism by which NKCC1-induced Cl(-) movement is autoregulated and by which Cl(-) entry and exit on opposite sides of polarized cells are coordinated. Although this signaling mechanism is coupled to a pathway that leads to post-translational modification of the carrier, no unifying model currently accounts for the ion dependence of NKCC1 regulation. In this paper, evidence is presented for the first time that hsp90 associates with the cytosolic C terminus of NKCC1, probably when the carrier is predominantly in its unfolded form during early biogenesis. Evidence is also presented that the Cl(-)(i)-dependent regulatory pathway can be activated by a thermal stress but that it is no longer operational if NKCC1-expressing cells are pretreated with geldanamycin, an antibiotic that inhibits hsp90, albeit nonspecifically. Taken together, our data indicate that binding of hsp90 to NKCC1 may be required for Na(+)-K(+)-Cl(-) cotransport to occur at the cell surface and that it could play an important role in ion-dependent signaling mechanisms, insofar as the maneuvers that were used to alter the expression or activity of the chaperone do not exert their main effect by inducing other cellular events such as the unfolded protein response. Further studies will be required to elucidate the functional relevance of this novel interaction. PMID- 15347683 TI - Tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma stimulate gamma-secretase-mediated cleavage of amyloid precursor protein through a JNK dependent MAPK pathway. AB - The deposition of the amyloid beta (Abeta) peptide in neuritic plaques plays a critical role in the pathogenesis of Alzheimer's disease (AD). Abeta is generated through the proteolysis of amyloid precursor protein (APP) by the sequential actions of beta- and gamma-secretases. Although recent evidence has unveiled much about the biochemical identity and characteristics of gamma-secretase, the mechanism regulating endogenous gamma-secretase activity remains elusive. To identify possible extracellular signals and associated signaling cascades that could regulate APP proteolysis by gamma-secretase activity, we have developed a cell-based reporter gene assay by stably cotransfecting HEK293 cells with the Gal4-driven luciferase reporter gene and the Gal4/VP16-tagged C-terminal fragment of APP (C99-GV), the immediate substrate of gamma-secretase. The cleavage of C99 GV by gamma-secretase releases the transcription factor that activates luciferase expression, providing a quantitative measurement of gamma-secretase activity. Using this reporter assay, we have demonstrated that interferon-gamma, interleukin-1beta, and tumor necrosis factor-alpha can specifically stimulate gamma-secretase activity, concomitant with increased production of Abeta and the intracellular domain of APP (AICD). The gamma-secretase-dependent cleavage of Notch is also enhanced upon the stimulation of these cytokines. The cytokine enhanced gamma-secretase activity can be suppressed by a potent inhibitor of c Jun N-terminal kinase (JNK). Furthermore, cells transfected with dominant positive MEKK1, one of the most potent activators of the JNK cascade, exhibit increased gamma-secretase activity, suggesting that the JNK-dependent mitogen activated protein kinase pathway could mediate the cytokine-elicited regulation of gamma-secretase. Our studies provide direct evidence that cytokine-elicited signaling cascades control Abeta production by modulating gamma-secretase activity. PMID- 15347684 TI - Amyloid-beta protein precursor (AbetaPP) intracellular domain-associated protein 1 proteins bind to AbetaPP and modulate its processing in an isoform-specific manner. AB - The amyloid-beta protein precursor (AbetaPP) is a type I transmembrane molecule that undergoes several finely regulated cleavage events. The physiopathological relevance of AbetaPP derives from the fact that its aberrant processing strongly correlates with the onset of Alzheimer's disease (AD). AD is a neurodegenerative disorder characterized by neuronal cell death, loss of synapses, and deposition of misfolded protein plaques in the brain; the main constituent of these plaques is the amyloid-beta peptide, a 40-42 amino-acid-long protein fragment derived by AbetaPP upon two sequential processing events. Mutations in the genes encoding for AbetaPP and some of the enzymes responsible for its processing are strongly associated with familial forms of early onset AD. Therefore, the elucidation of the mechanisms underlying AbetaPP metabolism appears crucial to understanding the basis for the onset of AD. Apart from Abeta, upon processing of AbetaPP other fragments are generated. The long extracellular domain is released in the extracellular space, whereas the short cytoplasmic tail, named AbetaPP intracellular domain (AID) is released intracellularly. AID appears be involved in several cellular processes, apoptosis, calcium homeostasis, and transcriptional regulation. We have recently reported the cloning and characterization of different isoforms of AID associated protein-1 (AIDA-1), a novel AID-binding protein. Here we further analyzed the interaction between several AIDA-1 isoforms and the cytoplasmic tail of AbetaPP. Our data demonstrated that the interaction between the two molecules is regulated by alternative splicing of the AIDA-1 proteins. Furthermore, we provide data supporting a possible function for AIDA-1a as a modulator of AbetaPP processing. PMID- 15347685 TI - The neuronal adaptor protein X11beta reduces amyloid beta-protein levels and amyloid plaque formation in the brains of transgenic mice. AB - Accumulation of cerebral amyloid beta-protein (Abeta) is believed to be part of the pathogenic process in Alzheimer's disease. Abeta is derived by proteolytic cleavage from a precursor protein, the amyloid precursor protein (APP). APP is a type-1 membrane-spanning protein, and its carboxyl-terminal intracellular domain binds to X11beta, a neuronal adaptor protein. X11beta has been shown to inhibit the production of Abeta in transfected non-neuronal cells in culture. However, whether this is also the case in vivo in the brain and whether X11beta can also inhibit the deposition of Abeta as amyloid plaques is not known. Here we show that transgenic overexpression of X11beta in neurons leads to a decrease in cerebral Abeta levels in transgenic APPswe Tg2576 mice that are a model of the amyloid pathology of Alzheimer's disease. Moreover, overexpression of X11beta retards amyloid plaque formation in these APPswe mice. Our findings suggest that modulation of X11beta function may represent a novel therapeutic approach for preventing the amyloid pathology of Alzheimer's disease. PMID- 15347686 TI - Dermatan sulfate proteoglycan and glycosaminoglycan synthesis is induced in fibroblasts by transfer to a three-dimensional extracellular environment. AB - Composition and architecture of the extracellular matrix dictate cell behavior. Proteoglycans bind multiple components of the extracellular matrix by serving as important regulators of cell behavior. Given the influence of culture architecture on cell function, we investigated whether switching NIH3T3 fibroblasts from growth on type 1 collagen in monolayer to a collagen gel might influence dermatan sulfate expression. Immunofluorescent staining, immunoblot, and Western blot demonstrated an induction in decorin expression in cells switched to collagen gels. This induction was associated with a 40-fold increase in decorin transcript expression determined by quantitative real time PCR. Disaccharide analysis of extracted glycosaminoglycans from collagen gels showed an increase in total glycosaminoglycan and in the ratio of chondroitin sulfate to heparan sulfate compared with monolayer culture. The ratio of chondroitin sulfate to heparan sulfate likewise increased on syndecan-1 from gel culture. Digestion with chondroitinase B showed that this induced chondroitin sulfate was dermatan sulfate. Syndecan-1 extracted from wounded mouse skin also displayed an increase in dermatan sulfate synthesis compared with unwounded skin. Furthermore, glycosaminoglycans from collagen gel culture activated keratinocyte growth factor, whereas glycosaminoglycans from monolayer culture lacked this ability. These findings suggest that regulation of dermatan sulfate and dermatan sulfate proteoglycan is dependent on extracellular matrix architecture. The ability of collagen gel culture to mimic better the in vivo dermal environment may be due in part to this influence on dermatan sulfate and dermatan sulfate proteoglycan synthesis. PMID- 15347687 TI - Glutathione S-transferase Pi has at least three distinguishable xenobiotic substrate sites close to its glutathione-binding site. AB - Benzyl isothiocyanate (BITC), present in cruciferous vegetables, is an efficient substrate of human glutathione S-transferase P1-1 (hGST P1-1). BITC also acts as an affinity label of hGST P1-1 in the absence of glutathione, yielding an enzyme inactive toward BITC as substrate. As monitored by using BITC as substrate, the dependence of k of inactivation (K(I)) of hGST P1-1 on [BITC] is hyperbolic, with K(I) = 66 +/- 7 microM. The enzyme incorporates 2 mol of BITC/mol of enzyme subunit upon complete inactivation. S-Methylglutathione and 8-anilino-1 naphthalene sulfonate (ANS) each yield partial protection against inactivation and decrease reagent incorporation, whereas S-(N-benzylthiocarbamoyl)glutathione or S-methylglutathione + ANS protects completely. Mapping of proteolytic digests of modified enzyme by using mass spectrometry reveals that Tyr(103) and Cys(47) are modified equally. S-Methylglutathione reduces modification of Cys(47), indicating this residue is at/near the glutathione binding region, whereas ANS decreases modification of Tyr(103), suggesting this residue is at/near the BITC substrate site, which is also near the binding site of ANS. The Y103F and Y103S mutant enzymes were generated, expressed, and purified. Both mutants handle substrate 1-chloro-2,4-dinitrobenzene normally; however, Y103S exhibits a 30-fold increase in K(m) for BITC and binds ANS poorly, whereas Y103F has a normal K(m) for BITC and K(d) for ANS. These results indicate that an aromatic residue at position 103 is essential for the binding of BITC and ANS. This study provides evidence for the existence of a novel xenobiotic substrate site in hGST P1-1, which can be occupied by benzyl isothiocyanate and is distinct from that of monobromobimane and 1-chloro-2,4 dinitrobenzene. PMID- 15347688 TI - Interaction with protocadherin-gamma regulates the cell surface expression of protocadherin-alpha. AB - The protocadherin-alpha (CNR/Pcdhalpha) and protocadherin-gamma (Pcdhgamma) proteins, members of the cadherin superfamily, are putative cell recognition/adhesion molecules in the brain. Overexpressed cadherins are generally expressed on the cell surface and elicit cell adhesion activity in several cell lines, although hardly any overexpressed CNR/Pcdhalpha proteins are expressed on the cell surface, except on HEK293T cells, which show low expression. We analyzed the expression of CNR/Pcdhalpha and Pcdhgamma in HEK293T cells and found that they formed a protein complex and that Pcdhgamma enhanced the surface expression of CNR/Pcdhalpha. This enhanced surface expression was confirmed by flow cytometry analysis and by marking cell surface proteins with biotin. The enhancement was observed using different combinations of CNR/Pcdhalpha and Pcdhgamma proteins. The surface expression activity was enhanced by the extracellular domains of the proteins, which could bind each other. Their cytoplasmic domains also had binding activity and influenced their localization. Their protein-protein interaction was also detected in extracts of mouse brain and two neuroblastoma cell lines. Thus, interactions between CNR/Pcdhalpha and Pcdhgamma regulate their surface expression and contribute to the combinatorial diversity of cell recognition proteins in the brain. PMID- 15347689 TI - The role of the conserved residues His-246, His-199, and Tyr-255 in the catalysis of catechol 2,3-dioxygenase from Pseudomonas stutzeri OX1. AB - Catechol 2,3-dioxygenase (C2,3O) from Pseudomonas stutzeri OX1, which is able to grow on various aromatic substrates as the sole source of carbon and energy, has been expressed in Escherichia coli, purified, characterized, and found to be very similar to other dioxygenases from Pseudomonas species. Interestingly, the activity of the protein shows a rather unusual pH dependence when assayed on catechol. A model of the catalytic mechanism was developed that is able to reproduce the catalytic behavior of the protein as a function of the pH. The model includes multiple equilibria and four productive intermediates with different ionization states of the enzyme-substrate complex. The fitting of the theoretical curve to the experimental data suggests that a tyrosine and two histidine residues are involved in catalysis. Mutants (H246N)-, (H246A)-, (H199N) and (Y255F)-C2,3O were produced to investigate the role of highly conserved His 199, His-246, and Tyr-255. The strongly reduced activity of the mutants suggests a primary catalytic role for each of these residues. Moreover, mutants at positions 199 and 246 display pH profiles different from that of the wild-type protein, thus indicating that residues His-246 and His-199 play a role in determining the unusual pH dependence of the enzyme. In addition, electron withdrawing groups on catechol, which increase the acidity of the phenolic hydroxyl group, are able to counterbalance the effect of the mutation H246N in reducing catalytic activity but cause a further reduction of the activity of (H199N)-C2,3O. This finding suggests that His-246 is involved in the initial catechol deprotonation, whereas His-199 promotes the reaction between oxygen and the aromatic ring. PMID- 15347690 TI - 14-3-3 protein C-terminal stretch occupies ligand binding groove and is displaced by phosphopeptide binding. AB - 14-3-3 proteins are important regulators of numerous cellular signaling circuits. They bind to phosphorylated protein ligands and regulate their functions by a number of different mechanisms. The C-terminal part of the 14-3-3 protein is known to be involved in the regulation of 14-3-3 binding properties. The structure of this region is unknown; however, a possible location of the C terminal stretch within the ligand binding groove of the 14-3-3 protein has been suggested. To fully understand the role of the C-terminal stretch in the regulation of the 14-3-3 protein binding properties, we investigated the physical location of the C-terminal stretch and its changes upon the ligand binding. For this purpose, we have used Forster resonance energy transfer (FRET) measurements and molecular dynamics simulation. FRET measurements between Trp242 located at the end of the C-terminal stretch and a dansyl group attached at two different cysteine residues (Cys25 or Cys189) indicated that in the absence of the ligand, the C-terminal stretch occupies the ligand binding groove of 14-3-3 protein. Our data also showed that phosphopeptide binding displaces the C-terminal stretch from the ligand binding groove. Intramolecular distances calculated from FRET measurements fit well with distances obtained from molecular dynamics simulation of full-length 14-3-3zeta protein. PMID- 15347691 TI - Protein stabilization by osmolytes from hyperthermophiles: effect of mannosylglycerate on the thermal unfolding of recombinant nuclease a from Staphylococcus aureus studied by picosecond time-resolved fluorescence and calorimetry. AB - 2-O-alpha-Mannosylglycerate, a negatively charged osmolyte widely distributed among (hyper)thermophilic microorganisms, is known to provide notable protection to proteins against thermal denaturation. To study the mechanism responsible for protein stabilization, pico-second time-resolved fluorescence spectroscopy was used to characterize the thermal unfolding of a model protein, Staphylococcus aureus recombinant nuclease A (SNase), in the presence or absence of mannosylglycerate. The fluorescence decay times are signatures of the protein state, and the pre-exponential coefficients are used to evaluate the molar fractions of the folded and unfolded states. Hence, direct determination of equilibrium constants of unfolding from molar fractions was carried out. Van't Hoff plots of the equilibrium constants provided reliable thermodynamic data for SNase unfolding. Differential scanning calorimetry was used to validate this thermodynamic analysis. The presence of 0.5 m potassium mannosylglycerate caused an increase of 7 degrees C in the SNase melting temperature and a 2-fold increase in the unfolding heat capacity. Despite the considerable degree of stabilization rendered by this solute, the nature and population of protein states along unfolding were not altered in the presence of mannosylglycerate, denoting that the unfolding pathway of SNase was unaffected. The stabilization of SNase by mannosylglycerate arises from decreased unfolding entropy up to 65 degrees C and from an enthalpy increase above this temperature. In molecular terms, stabilization is interpreted as resulting from destabilization of the denatured state caused by preferential exclusion of the solute from the protein hydration shell upon unfolding, and stabilization of the native state by specific interactions. The physiological significance of charged solutes in hyperthermophiles is discussed. PMID- 15347692 TI - X-ray imaging physics for nuclear medicine technologists. Part 1: Basic principles of x-ray production. AB - The purpose is to review in a 4-part series: (i) the basic principles of x-ray production, (ii) x-ray interactions and data capture/conversion, (iii) acquisition/creation of the CT image, and (iv) operational details of a modern multislice CT scanner integrated with a PET scanner. Advances in PET technology have lead to widespread applications in diagnostic imaging and oncologic staging of disease. Combined PET/CT scanners provide the high-resolution anatomic imaging capability of CT with the metabolic and physiologic information by PET, to offer a significant increase in information content useful for the diagnostician and radiation oncologist, neurosurgeon, or other physician needing both anatomic detail and knowledge of disease extent. Nuclear medicine technologists at the forefront of PET should therefore have a good understanding of x-ray imaging physics and basic CT scanner operation, as covered by this 4-part series. After reading the first article on x-ray production, the nuclear medicine technologist will be familiar with (a) the physical characteristics of x-rays relative to other electromagnetic radiations, including gamma-rays in terms of energy, wavelength, and frequency; (b) methods of x-ray production and the characteristics of the output x-ray spectrum; (c) components necessary to produce x-rays, including the x-ray tube/x-ray generator and the parameters that control x-ray quality (energy) and quantity; (d) x-ray production limitations caused by heating and the impact on image acquisition and clinical throughput; and (e) a glossary of terms to assist in the understanding of this information. PMID- 15347693 TI - Technical errors in planar bone scanning. AB - Optimal technique for planar bone scanning improves image quality, which in turn improves diagnostic efficacy. Because planar bone scanning is one of the most frequently performed nuclear medicine examinations, maintaining high standards for this examination is a daily concern for most nuclear medicine departments. Although some problems such as patient motion are frequently encountered, the degraded images produced by many other deviations from optimal technique are rarely seen in clinical practice and therefore may be difficult to recognize. The objectives of this article are to list optimal techniques for 3-phase and whole body bone scanning, to describe and illustrate a selection of deviations from these optimal techniques for planar bone scanning, and to explain how to minimize or avoid such technical errors. PMID- 15347694 TI - Impact of patient motion on myocardial perfusion SPECT diagnostic integrity: Part 2. AB - OBJECTIVE: Advances in 99mTc-based radiopharmaceuticals and multiple-detector gantries have the potential to increase the significance of patient motion on the diagnostic integrity of myocardial perfusion SPECT acquisitions. METHODS: An experimental study was used to evaluate the effect of various patient motions on the diagnostic integrity of myocardial perfusion SPECT data using 522 motion simulations generated from a technically and diagnostically normal dataset. RESULTS: Of studies with induced motion, 21.7% of simulated motion demonstrated motion-induced artifacts. Abrupt motion resulted in artifacts for 52.6% of studies, whereas bounce motion resulted in artifacts in 6.8% of studies. The locations where motion resulted in the most studies with artifacts were at 45 degrees (36.1%) and 75 degrees (32.4%). No statistical difference was demonstrated between single, dual-, and triple-head configurations. CONCLUSION: Combining these results with those of the clinical evaluation of incidence indicates that patient motion during 99mTc-based myocardial perfusion SPECT studies is a potential source of false-positive findings for coronary artery disease. There is a 7.1% probability that myocardial perfusion SPECT studies performed at the 3 sites investigated will contain a motion-induced artifact. Fully realized, this potential results in decreased test specificity and unfavorable cost and consequence outcomes. PMID- 15347695 TI - The helmet sign: Physiologic radioactive accumulation after 131I therapy--a case report. AB - After 131I radioablative treatment, a 51-y-old woman underwent whole-body (131)I scanning, which revealed intense uptake along the periphery of the skull. The patient disclosed that she had not washed her hair because she had obtained a new hairstyle between the 131I treatment and the scan. The intense uptake along the periphery of the skull represented radioactive physiologic accumulation at the patient's scalp. PMID- 15347696 TI - Distance assisted training in sub-Saharan Africa: a program evaluation. AB - Technologists working in nuclear medicine departments in sub-Saharan African countries do not have access to formal training in nuclear medicine and have been recruited mostly from related fields of radiologic technology. Because of the nature of the specialty, the numbers that require training are small, and it is therefore not cost-effective for higher-education institutions in these countries to set up training programs. There is also a lack of expertise in this field in Africa. Assessing the feasibility of running a distance assisted training program to provide training where none exists was undertaken as part of a project sponsored by the International Atomic Energy Agency and the African Co-Operative Agreement for Research, Development, and Training related to Nuclear Science and Technology. Seven countries were nominated, but only 3 centers in 2 countries, Sudan and Tanzania, had the infrastructure to support training. Twenty-one students received the first modules in November 1999, and 13 completed the course in December 2001. All students except one were examined in their own departments. Students received an IAEA Certificate of Achievement at the end of the course, at which time the program was evaluated. Analysis of the data indicated that the conceptualization and design of the material were excellent. There were, however, some problems with the implementation of the program, notably the lack of preparedness of the supervisors, limited departmental resources, and a range of nuclear medicine investigations inadequate for clinical competency. The course was seen to have a positive impact, as it not only allowed technologists to develop skills necessary for the profession but also encouraged critical thinking, reflection, and problem solving. One third of untrained nuclear medicine technologists working in sub-Saharan Africa have now received cost effective, structured on-site training. PMID- 15347697 TI - Family influences on pediatric asthma. AB - OBJECTIVE: To describe pathways by which families may influence the onset and course of a child's asthma. METHODS: We critically reviewed published articles and book chapters to identify research findings and integrated conceptualizations that demonstrate how families affect pediatric asthma. RESULTS: Family emotional characteristics, asthma management behaviors, and physiological factors account for key influences on pediatric asthma onset and outcomes. CONCLUSIONS: Multiple family characteristics are associated with pediatric asthma onset and outcomes. Behavioral and physiological mechanisms may act independently or may interact to affect asthma manifestations. Families with specific emotional characteristics may be at an elevated risk for poorer asthma outcomes. PMID- 15347698 TI - Memory matters: developmental differences in predictors of diabetes care behaviors. AB - OBJECTIVE: Longitudinal research shows that pediatric type 1 diabetes can result in verbal memory difficulties, yet the role of memory in the daily management of this or any other chronic illness has not been evaluated. METHODS: Verbal memory measures from two well-standardized tests were administered to 224 youths with type 1 diabetes, aged 9 to 17. Twenty-four-hour recall interviews conducted separately with mothers and their children assessed diabetes care behaviors. RESULTS: Rote verbal memory predicted blood glucose testing frequency for adolescents but not for preadolescents; and when combined with ethnicity, socioeconomic status, and age, rote verbal memory accounted for 27.6% of the variance, p <.001. Quantitative verbal working memory--along with ethnicity, socioeconomic status, and age, p <.01--accounted for 33.7% of the variance in predicting carbohydrate calories for older adolescents. CONCLUSIONS: Memory, in addition to demographic factors, is a significant predictor of some of the central self-care behaviors involved in diabetes management. However, memory only predicts diabetes management for older adolescents, who have greater self-care responsibility. PMID- 15347699 TI - Patterns of maternal distress among children with cancer and their association with child emotional and somatic distress. AB - OBJECTIVE: To identify patterns of distress among mothers of children with cancer over the initial 6 months of treatment and to examine these patterns as predictors of child somatic and emotional distress. METHOD: Data were gathered regarding maternal perceived stress and affective distress from mothers of children (N = 65, mean age = 8.3 years) with cancer at 2 to 5 weeks postdiagnosis, then at 12 to 14 weeks and 22 to 24 weeks. Mothers and nurses provided indexes of child somatic and emotional distress at these assessments. RESULTS: Hierarchical and k-means cluster analyses revealed four distinct patterns of maternal distress: high, moderate, declining, and low. The high maternal distress group reported higher child emotional distress at all three points but higher child somatic distress only at the final assessment. Maternal distress group was unrelated to nurse-reported child distress. CONCLUSIONS: The identification of four empirically derived patterns of maternal distress may explain some of the variance in the literature regarding parental distress vis-a vis pediatric cancer treatment and may have relevance to intervention efforts. Differences in the relations between maternal distress groups and mother- and nurse-reported child distress underscore the importance of collecting child distress data from multiple sources. PMID- 15347700 TI - Inpatient treatment for children with obesity: weight loss, psychological well being, and eating behavior. AB - OBJECTIVE: Evaluated the effects of a 10-month inpatient treatment program and implemented as a nondiet healthy lifestyle approach. In addition, the effects of two extended treatment programs were compared to a standard cognitive-behavioral treatment program for maintenance of the treatment gains. METHODS: A within subjects design was employed to evaluate treatment outcome, including a 14-month follow-up. Children (N = 122) ranged in age from 7 to 17 years (M = 12.7 years) with a mean Body Mass Index (BMI) of 32.5, expressed as a percentage of overweight (M = 77.4%). RESULTS: The children lost 49.0% of their weight during the course of treatment. Comparing baseline with the 14-month follow-up, a weight loss of 31.7% was maintained. The children continued to show healthy eating behavior at follow-up, and their psychological well-being had improved. No significant interaction effects were found for the extended coping programs. CONCLUSION: An inpatient cognitive-behavioral nondiet approach is a promising treatment option for childhood obesity, with lasting effects throughout the 14 month posttreatment. PMID- 15347701 TI - Cancer-related parental beliefs: the Family Illness Beliefs Inventory (FIBI). AB - OBJECTIVE: The goal of this study was to develop a reliable and valid method for assessing the cancer-related beliefs of parents with a child in treatment for cancer. METHOD: One hundred twenty-five families (119 mothers, 56 fathers) completed a measure of cancer-related beliefs written to reflect common themes associated with the diagnosis and treatment of childhood cancer. Participants also completed self-report questionnaires used for validation of the Family Illness Beliefs Inventory (FIBI). RESULTS: Principal-components analysis was used to derive a 41-item five-factor solution from the maternal data--Factor 1: Treatment-Related Suffering; Factor 2: Death and Devastation; Factor 3: Caregiver Competence; Factor 4: Connection; and Factor 5: Finding Meaning. Correlations with validation measures supported the factor structure. Paternal data showed similar patterns. CONCLUSIONS: The FIBI is a psychometrically sound method for identifying parental cancer-related beliefs. This measure may be helpful in developing and evaluating interventions to reduce parental distress related to childhood cancer and promote adaptive family functioning. PMID- 15347703 TI - Prediction of function from infancy to early childhood: implications for pediatric psychology. AB - OBJECTIVE: To determine whether item groupings derived from the Bayley Infant Neurodevelopmental Screener (BINS) are stable and predictive of 36-month cognitive and motor outcome. METHODS: BINS was administered at 6, 12, and 24 months, and the Bayley-II or McCarthy scales at 36 months. The BINS was factor analyzed, and factors, biomedical and environmental variables, were related to 36 month outcomes. RESULTS: Three factors were identified at each age, accounting for 52% to 64% of the variance. Continuity in factors over infancy and predictive utility of similar functions at 36 months were found. Optimal factor scores (> or = 75th percentile) increased the likelihood of later normal cognitive or motor outcome (ORs 2.14-7.94). CONCLUSIONS: Stability and continuity over time exist in specific subdomains of function on a neurodevelopmental screening test. PMID- 15347702 TI - Expressive and receptive language functioning in preschool children with prenatal cocaine exposure. AB - OBJECTIVE: To estimate the relationship between severity of prenatal cocaine exposure and expressive and receptive language skills in full-term, African American children at age 3 years. METHODS: Language was assessed at age 3 using the Clinical Evaluation of Language Fundamentals-Preschool (CELF-P). The sample included 424 children (226 cocaine exposed, 198 non-cocaine exposed) who received preschool language assessments at age 3, drawn from a cohort of 476 children enrolled prospectively at birth. RESULTS: Structural equation modeling was used to regress expressive and receptive language as intercorrelated response variables on level of prenatal cocaine exposure, measured by a latent construct including maternal self-report of cocaine use and maternal/infant urine toxicology assays and infant meconium. Results indicated a.168 SD decrease in expressive language functioning for every unit increase in exposure level (95% CI = -.320, -.015; p =.031) after consideration for fetal growth and gestational age as correlated response variables. Receptive language was more modestly related to prenatal cocaine exposure and was not statistically significant. Results for expressive language remained stable with inclusion of the McCarthy general cognitive index as a response variable (expressive language beta = -.173, 95% CI = -.330, -.016; p =.031), and with adjustment for maternal age and prenatal exposures to alcohol, tobacco, and marijuana (expressive language beta = -.175, 95% CI = -.347, -.003; p =.046). Additional child and caregiver environmental variables assessed at age 3 were also evaluated in varying statistical models with similar results. CONCLUSION: The evidence from this study supports a gradient relationship between increased level of prenatal cocaine exposure and decreased expressive language functioning in preschool-aged cocaine-exposed children. PMID- 15347704 TI - Brief report: Predictors of parenting stress among parents of children with biochemical genetic disorders. AB - OBJECTIVE: To examine predictors of parenting stress in parents whose children were diagnosed with a biochemical genetic disorder clinically or through newborn screening. METHODS: Parents of 263 children with biochemical genetic disorders (139 identified by newborn screening, 124 identified clinically) completed interviews focused on child health, medical service use, satisfaction with services, parenting stress, and family functioning. RESULTS: Multiple regression analyses suggested that child adaptive functioning, parental satisfaction with support, and difficulties parents experienced meeting their child's health care needs were associated with scores on the Parenting Stress Index (R2 =.51). CONCLUSIONS: Initiatives to improve child adaptive functioning and parental support as well as practical assistance to help parents meet their child's health needs may reduce parental stress and family disruption in this population. PMID- 15347705 TI - Commentary. Health and behavior CPT codes: an opportunity to revolutionize reimbursement in pediatric psychology. AB - Recently accepted codes in the Current Procedural Terminology system permit licensed pediatric psychologists to deliver psychological services utilizing billing codes that focus on improving medical and physical health. They potentially allow pediatric psychologists to obtain third-party reimbursement for services within a biopsychosocial model of care without requiring a concurrent psychiatric diagnosis. These codes represent a potential major paradigm shift whereby the services of a pediatric psychologist are provided and billed within the context of physical health care. Information is provided on the history of these codes along with detailed information regarding their utilization. Future directions for implementation, teaching, and research are provided along with a strong encouragement for use of these codes by pediatric psychologists. PMID- 15347706 TI - 18F-FDG PET in evaluating patients treated for metastatic colorectal cancer: can we predict prognosis? PMID- 15347707 TI - Understanding the standardized uptake value, its methods, and implications for usage. PMID- 15347708 TI - Molecular transport mechanisms of radiolabeled amino acids for PET and SPECT. PMID- 15347709 TI - PET for evaluation of differential myocardial perfusion dynamics after VEGF gene therapy and laser therapy in end-stage coronary artery disease. AB - The purpose of this study was to appraise the value of PET in the assessment of the effect of supposedly proangiogenic new therapies such as gene therapy with vascular endothelial growth factor (VEGF) gene and endomyocardial laser therapy. METHODS: Thirty-five patients with end-stage coronary artery disease and class III (Canadian Cardiovascular Society) angina were included. Myocardial ischemia was evaluated with dipyridamole PET scanning and exercise tolerance with bicycle ergometry. Ten patients were treated with naked plasmid DNA encoding for human VEGF165 (VEGF) and 12 patients were treated with laser therapy (direct myocardial revascularization [DMR]) using an electromechanical mapping system. Thirteen patients were treated with standard medical therapy (control). RESULTS: In both active treatment groups, angina was reduced in most subjects, except in 2 VEGF and 5 DMR patients. In the control group, no improvement in anginal classification was found, except in 3 subjects. On the PET scan, solely in the VEGF group, the stress perfusion was significantly improved (from 57 +/- 33 to 81 +/- 55 mL/min/100 g; P = 0.031). Furthermore, in the VEGF group, the number of ischemic segments was reduced from 274 +/- 41 to 234 +/- 48 segments (P = 0.004) but not in the DMR group (from 209 +/- 43 to 215 +/- 52 segments) or in the control group (from 218 +/- 18 to 213 +/- 28 segments). Bicycle exercise duration showed slight nonsignificant changes in the VEGF group (from 3.6 +/- 2.0 to 4.6 +/- 2.1 min), in the DMR group (from 5.1 +/- 1.5 to 4.7 +/- 1.3 min), and in the control group (from 3.3 +/- 1.8 to 3.5 +/- 2.3 min). CONCLUSION: PET showed that intramyocardial gene therapy with the human VEGF165 gene in contrast to laser DMR treatment effectively reduces myocardial ischemia. PMID- 15347710 TI - Superimposed display of coronary artery on gated myocardial perfusion scintigraphy. AB - Fusion of images of vascular anatomy and of myocardial perfusion images might be helpful for understanding the relationship between ischemia and the responsible vessels. The aim of this study was to develop a simple means of superimposing the images obtained from coronary angiography and gated myocardial perfusion SPECT. METHODS: Right and left oblique views from conventional coronary angiography and left ventriculography (LVG) were stored as 512 x 512 x 8-bit digital datasets and combined. We reconstructed images from routine gated myocardial perfusion imaging (MPI) by using (99m)Tc-tetrofosmin to match the oblique positions between the image from MPI and combined angiographic images. We then generated a 3 dimensional (3D) surface map by using the quantitative gated SPECT (QGS)/quantitative perfusion SPECT (QPS) program. Both the combined angiographic images and the 3D surface map were rescaled and unified by registering the internal landmarks between the 2 images. After subtraction of the LVG image, the coronary angiogram and the 3D surface map were fused into 1 image. All processes were performed with the QGS/QPS program and commercially available graphic software. We applied this method to datasets from a cardiac phantom and from several patients with coronary artery disease. RESULTS: In the phantom study, our technique could obtain a 3D surface map in which the oblique angle was identified as that of radiography and could realize image registration and superimposition of radiography on scintigraphy. The preliminary results from the patients indicated that the markedly stenotic vessels showed good coincidence with the regional myocardial perfusion abnormalities on the unified images. In addition, these images could show the relationship between the coronary artery and regional wall motion in the gated mode. CONCLUSION: We developed a simple method of superimposing the image of the coronary artery tree on images from gated MPI. The technique yielded useful information about myocardial perfusion and function as well as the supplying coronary artery. PMID- 15347711 TI - Superiority of 3-dimensional stereotactic surface projection analysis over visual inspection in discrimination of patients with very early Alzheimer's disease from controls using brain perfusion SPECT. AB - In Alzheimer's disease (AD), regional cerebral blood flow (rCBF) in the posterior cingulate gyri and precunei has been reported to decrease even at a very early stage. It may be helpful to use statistical image analysis to distinguish slight decreases in rCBF in this area. We compared a 3-dimensional stereotactic surface projection (3D-SSP) technique with visual inspection in the discrimination of patients with very early AD from age-matched controls using brain perfusion SPECT. METHODS: SPECT was obtained in 38 patients with probable AD at a very early stage and after a mean interval of 15 mo and in 76 age-matched healthy volunteers. We randomly divided these subjects into 2 groups. The first group was used to identify the areas with significant decreases of rCBF in patients compared with healthy control subjects based on the voxel-based analysis using 3D SSP. The second group was used to compare the discrimination ability between patients and control subjects by 3D-SSP with that by visual inspection. In the second group, a Z-score map for a SPECT image of a subject was obtained by comparison with mean and SD SPECT images of control subjects for each voxel after anatomic standardization and voxel normalization to reference regions. Receiver operating characteristic (ROC) curves for a Z-score discriminating patients with AD from control subjects were analyzed in areas with significant decreases of rCBF identified in the first group. For visual inspection, 6 physicians graded the rCBF decrease on SPECT images for ROC curves. They inspected the images twice at an interval of >2 wk, and intra- and interobserver reliabilities were determined. RESULTS: Visual inspection showed fair-to-excellent intra- and interobserver reliabilities. The 3D-SSP demonstrated an accuracy of 86.2% for discriminating patients with AD from control subjects when analyzing the posterior cingulate gyri and precunei with global mean normalization. In contrast, visual inspection did not show an accuracy of >74.0% for this discrimination. CONCLUSION: The ability of 3D-SSP to discriminate patients with very early AD from control subjects is superior to that of visual inspection. It is clinically useful and reliable to adopt the use of 3D-SSP as an adjunct to visual interpretation. PMID- 15347712 TI - Quantification of human nicotinic acetylcholine receptors with 123I-5IA SPECT. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are widely distributed in the human brain, especially the alpha4beta2 subtype of nAChR. The cholinergic systems have roles in various neurophysiologic functions, such as learning, memory, and cognition, whereas normal aging and neurodegenerative diseases have been associated with changes in nAChRs. Recently, 5-iodo-3-(2(S) azetidinylmethoxy)pyridine (5IA) has been synthesized as a radioligand for imaging nAChRs with SPECT. 123I-5IA shows higher affinity toward the nAChR alpha4beta2 subtype, enhanced receptor subtype selectivity, good safety, and low nonspecific binding. METHODS: In this study, a SPECT quantitative study of human nAChRs binding with 123I-5IA was conducted in healthy volunteers. An arterial input function was obtained for each subject and a 2-compartment model was used to determine the kinetic parameters of 123I-5IA using data from a 6-h scan. The distribution volume (VT (mL/mL), which is related to the number of unoccupied binding sites in the brain, was calculated and values were compared with results of a graphical analysis (Logan plot, VLG). RESULTS: Analysis of the unmetabolized compound showed a high parent fraction of 123I-5IA in plasma. The results from the 2-compartment model analysis showed high VT values for the thalamus; moderate values for the brain stem, cerebellum, and basal ganglia; and low values for the cortical regions. Good agreement was observed between VT values and results of autoradiographic experiments done in vitro for nAChR density in human brain. A high correlation index was observed between distribution volumes from model and graphical analyses. CONCLUSION: Our results indicated that 123I-5IA SPECT is suitable for the quantification of nAChRs in human brain. PMID- 15347713 TI - Brain incorporation of 11C-arachidonic acid, blood volume, and blood flow in healthy aging: a study with partial-volume correction. AB - PET with 11C-arachidonic acid (AA) can be used to quantify neural signaling related to phospholipase A2 (PLA2). Animal studies suggest reduction in the activity of this signaling system with age. The aim of this study was to evaluate the effect of healthy aging on brain incorporation of 11C-AA, before and after partial-volume correction (PVC). METHODS: Absolute measurements of cerebral blood flow (CBF) were obtained in 8 young and 7 old healthy subjects (mean age +/- SD, 27 +/- 5 y and 65 +/- 9 y) with bolus injection of 15O-water. About 15 min later, dynamic 60-min 3-dimensional scans were acquired after the injection of 11C-AA. Radioactivity frames of 11C-AA were corrected for head motion and registered to magnetic resonance (MR) images. A 3-segment (3S) and a 2-segment (2S) PVC was applied pixel-by-pixel to the activity frames. For the 3S method, the white matter value was estimated using a new automatic method by extrapolating the activity values of pixels with white matter membership > 0.99. Parametric images of the brain incorporation rate of 11C-AA (K*) and cerebral blood volume (Vb), as well as CBF, were generated and regional gray matter values were obtained. RESULTS: Among cortical areas, there were no significant differences (uncorrected P < 0.05) in K* or Vb absolute values between young and old subjects before or after PVC. A significant reduction of CBF was detected in the frontal cortex of the elderly group. After normalization to the global gray average, K*, Vb, and CBF values revealed significant reductions in the frontal lobe of old subjects; none of these differences were significant after PVC. CONCLUSION: These results confirm previous PET findings that brain function at rest is minimally affected by healthy aging. Proper PVC methodology is of critical importance in accurate quantitative assessment of PET physiologic measures. PMID- 15347714 TI - Prognostic aspects of 18F-FDG PET kinetics in patients with metastatic colorectal carcinoma receiving FOLFOX chemotherapy. AB - We evaluated quantitative measurement series (MS) with 18F-FDG and PET and compared different quantification methods for prediction of individual survival in patients with metastatic colorectal cancer receiving chemotherapy with 5 fluorouracil, folinic acid, and oxaliplatin (FOLFOX). METHODS: The study comprised 25 patients. All patients were examined before the onset of FOLFOX therapy and after completion of the first and fourth cycles. SUV, fractal dimension (FD), a 2-compartment model with computation of k1, k2, k3, and k4, and vascular fraction (VB) were used for data evaluation. Survival data served as a reference for the PET data. Discriminant analysis (DA), regression, and best subset analysis were applied to the data. RESULTS: Twenty of 25 patients died up to 801 d after the first PET study. A cutoff of 1 y (364 d) was used to classify the patients into 2 a priori groups, namely the short- and long-term survival groups. DA was used to predict the 2 categories using SUV and kinetic parameters of 18F-FDG metabolism as predictor variables. SUV provided a correct classification rate (CCR) ranging from 62% to 69%. SUV of the third MS resulted in a CCR of 69% as a single parameter. The best results were yielded by the use of kinetic parameters (k1, k3, VB, and FD) as predictor variables. CCR was 78% using kinetic 18F-FDG parameters of the first and third MS, in comparison with 69% for the corresponding SUVs. A multiple linear regression model was applied to the data to assess the relationship between individual survival and the PET data. The best-subset method revealed a correlation coefficient of 0.850 for the kinetic parameters of the first (k3, k4, VB, and FD) and third (k1, k2, k4, and VB) MS. CONCLUSION: The combination of kinetic parameters of the first and the third MS is acceptable for classification into a short or long survival class. Furthermore, even an individual prognosis of survival can be achieved using kinetic 18F-FDG parameters of the first and third MS. PMID- 15347715 TI - Ursodeoxycholic acid-augmented hepatobiliary scintigraphy in the evaluation of neonatal jaundice. AB - Early differentiation of extrahepatic biliary atresia from intrahepatic cholestasis is important. Hepatobiliary scintigraphy is an excellent noninvasive investigation for ruling out extrahepatic biliary atresia. This study aimed at identifying the role of ursodeoxycholic acid (UDCA), a choleretic agent, in conjunction with hepatobiliary scintigraphy in differentiating extrahepatic biliary atresia from neonatal hepatitis. METHODS: Fifty-one infants (42 male, 9 female) aged 0.3-5.5 mo (mean, 2.9 mo) presenting with neonatal jaundice underwent 99mTc-mebrofenin hepatobiliary scintigraphy. For patients who did not show any excretion of tracer into the intestine till 24 h, the study was repeated after oral administration of UDCA (20 mg/kg every 12 h) for 48-72 h. Ultrasonography and, if required, liver biopsy and intraoperative cholangiography were used with clinical data such as stool color and serologic and other etiologic investigations to form a final diagnosis. RESULTS: Of 51 patients, 19 showed biliary excretion in the first study, ruling out extrahepatic biliary atresia. Neonatal hepatitis was the final diagnosis in these. Of the remaining 32 patients, 12 nonexcretors converted to excretors after UDCA treatment, whereas 20 still showed no biliary drainage. Four nonexcretors on scintigraphy had a final diagnosis of neonatal hepatitis with galactosemia; the remaining 16 had extrahepatic biliary atresia. The specificity of hepatobiliary scintigraphy in ruling out extrahepatic biliary atresia improved from 54.3% to 88.6% (P < 0.001) after UDCA treatment. None of the patients experienced any ill effects from UDCA administration. CONCLUSION: Pretreatment with UDCA significantly improves the specificity of hepatobiliary scintigraphy in ruling out extrahepatic biliary atresia as a cause of prolonged neonatal jaundice. PMID- 15347716 TI - Comparison of different quantitative approaches to 18F-fluoride PET scans. AB - The aim of this study was to evaluate the relationship between 18F-fluoride bone metabolic measures obtained by nonlinear regression (NLR), Patlak analysis, and standardized uptake value (SUV) for a wide range of normal and pathologic bone conditions. In patients imaged twice, changes in metabolic rates were determined using the different quantitation methods. METHODS: In 33 patients 2-51 mo after resection of a bone tumor of the limbs, a total of 42 dynamic PET scans were performed after injection of 250-350 MBq 18F-fluoride. SUV, fluoride bone influx rate obtained by Patlak analysis (KPat), and fluoride bone influx rate obtained by NLR (KNLR) were calculated in each patient for the bone graft, the contralateral normal side, and the spine, if within the field of view. RESULTS: SUV ranged from 0.4 to 9.9, KPat from 0.0035 to 0.0742 mL/min/mL, and KNLR from 0.0027 to 0.0737 mL/min/mL. Significant linear correlations were found between KPat and KNLR (r = 0.99), KPat and SUV (r = 0.95), and KNLR and SUV (r = 0.93). Eight patients have been imaged twice within 6 mo. Changes in metabolic values of the grafts were significantly correlated for KPat and KNLR (r = 0.96), KPat and SUV (r = 0.88), and KNLR and SUV (r = 0.79). The 95% ranges of normal change in limb bones were +/-58.0% for SUV, +/-23.0% for KPat, and +/-20.2% for KNLR; the corresponding 95% ranges in the spine were +/-8.6%, +/-7.6%, and +/-19.6%. CONCLUSION: The results of this study show that 18F-fluoride metabolic values as well as changes in bone metabolism measured by SUV and Patlak analysis were strongly correlated with NLR findings. The high 95% range of normal change of SUV in limb bones, however, indicates that this parameter is of limited value in areas with low metabolic activity. The range of spontaneous bone metabolic rate fluctuations presented in this study may be used as an estimate for assessing changes in bone metabolic activity, and the normal values for limb bones provide a basis for further studies on 18F-fluoride bone metabolism. PMID- 15347717 TI - Tomographic imaging in the diagnosis of pulmonary embolism: a comparison between V/Q lung scintigraphy in SPECT technique and multislice spiral CT. AB - Although ventilation/perfusion (V/Q) lung scintigraphy is a well-accepted and frequently performed procedure in the diagnosis of pulmonary embolism, there is growing controversy about its relevance, particularly due to the increasing competition between scintigraphy and CT. Even though comparative studies between both modalities have already been performed, their results were highly inconsistent. Remarkably, in most of those studies, conventional planar perfusion scans were compared with tomographic images acquired using state-of-the-art CT scanners-a study design that cannot give impartial results. Hence, the aim of our study was a balanced comparison between V/Q lung scintigraphy and CT angiography using advanced imaging techniques for both modalities. METHODS: A total of 83 patients with suspected pulmonary embolism were examined using V/Q lung scintigraphy in SPECT technique as well as 4-slice spiral CT. Ventilation scans were done using an ultrafine aerosol. Additionally, planar images in 8 views were extracted from the V/Q SPECT datasets. Two experienced referees assessed each of the 3 modalities. The final diagnosis was made at a consensus meeting while taking into account all of the imaging modalities, laboratory tests, clinical data, and evaluation of a follow-up period. RESULTS: In the course of the consensus conference, pulmonary embolism was diagnosed in 37 of the 83 patients (44.6%). Compared with planar scintigraphy, SPECT raised the number of detectable defects at the segmental level by 12.8% (+11 defects; P = 0.401) and at the subsegmental level by 82.6% (+57 defects; P < 0.01). The sensitivity/specificity/accuracy of planar V/Q scintigraphy and V/Q SPECT was 0.76/0.85/0.81 and 0.97/0.91/0.94, respectively, compared with 0.86/0.98/0.93 for multislice CT. CONCLUSION: SPECT and ultrafine aerosols are technical advancements that can substantially improve lung scintigraphy. Using advanced imaging techniques, V/Q scintigraphy and multislice spiral CT both yield an excellent and, in all aspects, comparable diagnostic accuracy, with CT leading in specificity while SPECT shows a superior sensitivity. Even though planar lung scintigraphy yields satisfactory results for a nontomographic modality, it does not compare with tomographic imaging. PMID- 15347718 TI - Current concepts in lymph node imaging. AB - The accurate identification and characterization of lymph nodes by imaging has important therapeutic and prognostic significance in patients with newly diagnosed cancers. The presence of nodal metastases limits the therapeutic options and also generally indicates worse prognosis in patients. Thus, it becomes crucial to have this information before commencing therapy. Current cross sectional imaging modalities rely on insensitive size and morphologic criteria and, thus, lack the desired accuracy for characterizing lymph nodes. This is mainly because metastases can be present in non-enlarged lymph nodes and not all enlarged nodes are malignant. PET has overcome some of these limitations but is still constrained by current resolution limits for small nodal metastases. This has fueled the development of targeted techniques for nodal imaging and characterization as outlined in this article. In the past few years, studies have shown that these newer imaging techniques can bridge some of the limitations of existing imaging for nodal characterization and thereby provide the much-needed staging information before the initiation of therapy. PMID- 15347719 TI - Effects of noise, image resolution, and ROI definition on the accuracy of standard uptake values: a simulation study. AB - Semiquantitative standard uptake values (SUVs) are used for tumor diagnosis and response monitoring. However, the accuracy of the SUV and the accuracy of relative change during treatment are not well documented. Therefore, an experimental and simulation study was performed to determine the effects of noise, image resolution, and region-of-interest (ROI) definition on the accuracy of SUVs. METHODS: Experiments and simulations are based on thorax phantoms with tumors of 10-, 15-, 20-, and 30-mm diameter and background ratios (TBRs) of 2, 4, and 8. For the simulation study, sinograms were generated by forward projection of the phantoms. For each phantom, 50 sinograms were generated at 3 noise levels. All sinograms were reconstructed using ordered-subset expectation maximization (OSEM) with 2 iterations and 16 subsets, with or without a 6-mm gaussian filter. For each tumor, the maximum pixel value and the average of a 50%, a 70%, and an adaptive isocontour threshold ROI were derived as well as with an ROI of 15 x 15 mm. The accuracy of SUVs was assessed using the average of 50 ROI values. Treatment response was simulated by varying the tumor size or the TBR. RESULTS: For all situations, a strong correlation was found between maximum and isocontour based ROI values resulting in similar dependencies on image resolution and noise of all studied SUV measures. A strong variation with tumor size of > or =50% was found for all SUV values. For nonsmoothed data with high noise levels this variation was primarily due to noise, whereas for smoothed data with low noise levels partial-volume effects were most important. In general, SUVs showed under- and overestimations of > or =50% and depended on all parameters studied. However, SUV ratios, used for response monitoring, were only slightly dependent of ROI definition but were still affected by noise and resolution. CONCLUSION: The poor accuracy of the SUV under various conditions may hamper its use for diagnosis, especially in multicenter trials. SUV ratios used to measure response to treatment, however, are less dependent on noise, image resolution, and ROI definition. Therefore, the SUV might be more suitable for response-monitoring purposes. PMID- 15347720 TI - 18F-FDG kinetics and gene expression in giant cell tumors. AB - 18F-FDG kinetics were evaluated by use of compartment and noncompartment models of giant cell tumors. The kinetic data were compared with the gene expression data for a subgroup of patients. METHODS: Nineteen patients with giant cell tumors were examined with PET and 18F-FDG, and tracer kinetics were assessed quantitatively. A 2-compartment model, including the transport constants k1-k4 as well as the vascular fraction (VB) for 18F-FDG, was used for evaluation of the data. A noncompartment model was used to calculate the fractal dimension of the 18F-FDG time-activity curve to assess the heterogeneity of the tracer kinetics. Furthermore, tumor specimens obtained from 5 patients were assessed with gene chip technology (U95A), and these data were compared with the quantitative 18F FDG data. RESULTS: The giant cell tumors showed generally enhanced 18F-FDG uptake 1 h after tracer application, with a mean 18F-FDG standardized uptake value (SUV) of 4.8 (range, 1.8-9.4). Quantitative evaluation of tracer kinetics showed a preferential increase for 18F-FDG transport, with a mean k1 of 0.340. The vascular fraction accounted for 35% of the tumor volume and was high compared with those for other tumors, such as soft-tissue sarcomas. 18F-FDG kinetics were heterogeneous, with a fractal dimension of 1.3. Gene chip analysis showed that the expression of 137 genes (1.1%) exceeded the median expression value of the reference gene, beta2-microglobulin. The highest expression was observed for the gene for the small, leucine-rich proteoglycan I (biglycan), which is important for bone cell differentiation and proliferative activity. Correlation analysis revealed an association of 18F-FDG data with the expression of several genes. Mainly genes related to angiogenesis were associated with the compartment parameters. The SUV at 56-60 min was correlated with the expression of vascular endothelial growth factor A (angiogenesis) and cell division cycle 2 protein (proliferation). CONCLUSION: Despite their classification as benign tumors, giant cell tumors have generally enhanced 18F-FDG uptake, mainly attributable to an enhanced vascular fraction and increased 18F-FDG transport. A comparison of gene chip data and 18F-FDG kinetic data showed a close association of quantitative 18F FDG results and the expression of genes related to angiogenesis. PMID- 15347721 TI - Detection of cardiomyocyte death in a rat model of ischemia and reperfusion using 99mTc-labeled annexin V. AB - There is increasing evidence that cell death after myocardial ischemia and reperfusion may begin as apoptosis rather than necrosis. To determine the time course, location, and extent of this process, we studied groups of rats after a 20-min interval of coronary occlusion and reperfusion. METHODS: After thoracotomy, the left coronary artery was occluded for 20 min. After release and before study, groups of animals were allowed to recover for various intervals: 0.5 h (n = 6), 1.5 h (n = 7), 6 h (n = 7), 1 d (n = 8), 3 d (n = 8), or 2 wk (n = 5). At the time of study, the rats were injected with 99mTc-annexin V (80-150 MBq). One hour later, to verify the area at risk, 201Tl (0.74 MBq) was injected intravenously just after the left coronary artery reocclusion and the rats were sacrificed 1 min later. Dual-tracer autoradiography was performed to assess 99mTc annexin V uptake and the area at risk. RESULTS: Extensive 99mTc-annexin V uptake was observed in the mid myocardium after 0.5-1.5 h of reperfusion. The area of annexin uptake had expanded in the subendocardial and subepicardial layers at 6 h after reperfusion and then gradually lessened over 3 d. At 0.5 and 1.5 h of reperfusion, 99mTc-annexin V uptake ratios were 7.36 +/- 2.95 and 6.34 +/- 2.24 (mean +/- SD), respectively. The uptake ratios gradually decreased at 6 h, 1 d, 3 d, and 2 wk after reperfusion (4.65 +/- 1.93, 3.27 +/- 0.92 [P < 0.01 vs. 0.5 h], 1.84 +/- 0.55 [P < 0.001 vs. 0.5 h, P < 0.005 vs. 1.5 h], and 1.65 +/- 0.31 [P < 0.001 vs. 0.5 h, P < 0.005 vs. 1.5 h], respectively). CONCLUSION: These data indicate that annexin binding commences soon after ischemia and reperfusion in the mid myocardium within the area at risk and expands to include the subendocardial and subepicardial layers at 6 h after reperfusion, followed by gradual reduction of activity over 3 d. PMID- 15347722 TI - Radiation therapy of small cell lung cancer with 177Lu-DOTA-Tyr3-octreotate in an animal model. AB - Small cell lung cancer (SCLC) is a tumor of neuroendocrine (NE) origin with very low survival rate. Somatostatin receptor scintigraphy using 111In-DTPA-octreotide (DTPA is diethylenetriaminepentaacetic acid) is a well-established method for the visualization of somatostatin receptor-expressing NE tumors. Recently, new combinations of radionuclides and somatostatin analogs have been investigated for therapeutic purposes. In this study, the somatostatin analog DOTA-Tyr3-octreotate (DOTA is 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid), labeled with the medium-energy electron emitter 177Lu (maximal electron energy = 498 keV, half-life = 6.6 d), was used for radiation therapy of human SCLC in an animal model. METHODS: Nude mice, bearing tumors from the human SCLC cell line NCI-H69, were injected intravenously with 177Lu-DOTA-Tyr3-octreotate. Groups of animals (n = 5 or 6) were injected with 45-, 60-, and 120-MBq fractions and two 45-MBq fractions 48 h apart. Furthermore, 1 control group was treated with unlabeled DOTA-Tyr3-octreotate and another control group was not treated. RESULTS: In both control groups, the tumor volumes were increased 2-fold in approximately 5 d. Treatment with 177Lu-DOTA-Tyr3-octreotate resulted in marked tumor regression with statistically significant tumor volume reduction after 1 wk (P < 0.001). The tumor growth delay time was dependent on the amount of injected activity for the groups with single injections, 26 d for 60 MBq and 40 d for 120 MBq. The best therapeutic effect was obtained in mice injected with 2 fractions of 45 MBq. The relative tumor volume after 1 mo was 0.004 +/- 0.004. CONCLUSION: Radiation therapy with 177Lu-DOTA-Tyr3-octreotate on SCLC-bearing mice was successful. Since the experiments were performed on a human SCLC cell line xenografted to nude mice, the results may be clinically relevant and treatment with 177Lu-DOTA Tyr3-octreotate could be a treatment alternative in this tumor disease that normally has a dismal prognosis. PMID- 15347724 TI - Biodistribution and radiation dosimetry of the serotonin transporter ligand 11C DASB determined from human whole-body PET. AB - 11C-Labeled 3-amino-4-(2-dimethylaminomethylphenylsulfanyl)-benzonitrile (DASB) is a selective radioligand for the in vivo quantitation of serotonin transporters (SERTs) using PET. The goal of this study was to provide dosimetry estimates for 11C-DASB based on human whole-body PET. METHODS: Dynamic whole-body PET scans were acquired for 7 subjects after the injection of 669 +/- 97 MBq (18.1 +/- 2.6 mCi) of 11C-DASB. The acquisition for each subject was obtained at 14 time points for a total of 115 min after injection of the radioligand. Regions of interest were placed over compressed planar images of source organs that could be visually identified to generate time-activity curves. Radiation burden to the body was calculated from residence times of these source organs using the MIRDOSE3.1 program. RESULTS: The organs with high radiation burden included the lungs, urinary bladder wall, kidneys, gallbladder wall, heart wall, spleen, and liver. The activity peaked within 10 min after the injection of 11C-DASB for all these organs except two--the excretory organs gallbladder and urinary bladder wall, which had peak activities at 32 and 22 min, respectively. Monoexponential fitting of activity overlying the urinary bladder suggested that approximately 12% of activity was excreted via the urine. Simulations in which the urinary voiding interval was decreased from 4.8 to 0.6 h produced only modest effects on the dose to the urinary bladder wall. With a 2.4-h voiding interval, the calculated effective dose was 6.98 microGy/MBq (25.8 mrem/mCi). CONCLUSION: The estimated radiation burden of 11C-DASB is relatively modest and would allow multiple PET examinations of the same research subject per year. PMID- 15347723 TI - Bone marrow dosimetry and safety of high 131I activities given after recombinant human thyroid-stimulating hormone to treat metastatic differentiated thyroid cancer. AB - Recombinant human thyroid-stimulating hormone (rhTSH) recently was introduced as a radioiodine administration adjunct that avoids levothyroxine (LT-4) withdrawal and resultant hypothyroidism. The pharmacokinetics of 131I after rhTSH administration are known to differ from those after LT-4 withdrawal but are largely nondelineated in the radioiodine therapy setting. We therefore sought to calculate the red marrow absorbed dose of high therapeutic activities of 131I given after rhTSH administration to patients with metastatic or inoperable locally recurrent differentiated thyroid cancer. We also sought to evaluate the clinical and laboratory effects of this therapy on the bone marrow. METHODS: Fourteen consecutive patients received in total 17 131I treatments (7.4 GBq). Blood and urine samples were obtained at fixed intervals, and their activities were measured in a well counter. Based on blood activity, renal clearance of the activity, and residence times in red marrow and the remainder of the body, the red marrow absorbed dose was calculated using the MIRD schema. Additionally, we monitored for potential hematologic toxicity and compared platelet counts before and 3 mo after treatment. RESULTS: The mean +/- SD absorbed dose per unit of administered (131)I in the red marrow was 0.16 +/- 0.07 mGy/MBq. The corresponding total red marrow absorbed dose was 1.15 +/- 0.52 Gy (range, 0.28 1.91 Gy). In none of the patients was hematologic toxicity observed. The mean +/- SD platelet count (n = 13 treatments) was 243 +/- 62 x 10(9)/L before treatment and 233 +/- 87 x 10(9)/L 3 mo later, a slight and statistically insignificant decrease. After rhTSH-aided administration of high activities of 131I, the bone marrow absorbed dose remained under 2 Gy, the level long considered the safety threshold for all radioiodine therapy. CONCLUSION: Our specific findings imply that when clinically warranted, rhTSH should allow an increase in the therapeutic radioiodine activity. Such an increase might improve efficacy while preserving safety and tolerability; this possibility should be assessed in further studies. PMID- 15347725 TI - A tracer kinetic model for 18F-FHBG for quantitating herpes simplex virus type 1 thymidine kinase reporter gene expression in living animals using PET. AB - Reporter probe 9-(4-18F-fluoro-3-[hydroxymethyl]butyl)guanine (18F-FHBG) and reporter gene mutant herpes simplex virus type 1 thymidine kinase (HSV1-sr39tk) have been used for imaging reporter gene expression with PET. Current methods for quantitating the images using the percentage injected dose per gram of tissue do not distinguish between the effects of probe transport and subsequent phosphorylation. We therefore investigated tracer kinetic models for 18F-FHBG dynamic microPET data and noninvasive methods for determining blood time-activity curves in an adenoviral gene delivery model in mice. METHODS: 18F-FHBG (approximately 7.4 MBq [approximately 200 microCi]) was injected into 4 mice; 18F FHBG concentrations in plasma and whole blood were measured from mouse heart left ventricle (LV) direct sampling. Replication-incompetent adenovirus (0-2 x 10(9) plaque-forming units) with the E1 region deleted (n = 8) or replaced by HSV1 sr39tk (n = 18) was tail-vein injected into mice. Mice were dynamically scanned using microPET (approximately 7.4 MBq [approximately 200 microCi] 18F-FHBG) over 1 h; regions of interest were drawn on images of the heart and liver. Serial whole blood 18F-FHBG concentrations were measured in 6 of the mice by LV sampling, and 1 least-squares ratio of the heart image to the LV time-activity curve was calculated for all 6 mice. For 2 control mice and 9 mice expressing HSV1-sr39tk, heart image (input function) and liver image time-activity curves (tissue curves) were fit to 2- and 3-compartment models using Levenberg-Marquardt nonlinear regression. The models were compared using an F statistic. HSV1-sr39TK enzyme activity was determined from liver samples and compared with model parameter estimates. For another 3 control mice and 6 HSV1-sr39TK-positive mice, the model-predicted relative percentage of metabolites was compared with high performance liquid chromatography analysis. RESULTS: The ratio of 18F-FHBG in plasma to whole blood was 0.84 +/- 0.05 (mean +/- SE) by 30 s after injection. The least-squares ratio of the heart image time-activity curve to the LV time activity curve was 0.83 +/- 0.02, consistent with the recovery coefficient for the partial-volume effect (0.81) based on independent measures of heart geometry. A 3-compartment model best described 18F-FHBG kinetics in mice expressing HSV1 sr39tk in the liver; a 2-compartment model best described the kinetics in control mice. The 3-compartment model parameter, k3, correlated well with the HSV1-sr39TK enzyme activity (r2 = 0.88). CONCLUSION: 18F-FHBG equilibrates rapidly between plasma and whole blood in mice. Heart image time-activity curves corrected for partial-volume effects well approximate LV time-activity curves and can be used as input functions for 2- and 3-compartment models. The model parameter k3 from the 3-compartment model can be used as a noninvasive estimate for HSV1-sr39TK reporter protein activity and can predict the relative percentage of metabolites. PMID- 15347726 TI - Establishment of a human hepatocellular carcinoma cell line highly expressing sodium iodide symporter for radionuclide gene therapy. AB - To evaluate the possibility of radionuclide gene therapy and imaging in hepatocellular carcinoma cancer, we investigated the iodine accumulation of a human hepatocellular carcinoma cell line, SK-Hep1, by transfer of human sodium iodide symporter (hNIS) gene. By targeting NIS expression in SK-Hep1, we could also investigate whether these cells concentrate 99mTc-pertechnetate and 188Re perrhenate as well as 125I in vitro and in vivo. METHODS: The hNIS gene was transfected to human hepatocellular carcinoma SK-Hep1 cell lines using lipofectamine plus reagent. The uptake and efflux of 125I, 99mTc-pertechnetate, and 188Re-perrhenate were measured in the transfected and parental cells. Biodistribution was studied in nude mice bearing SK-Hep1 and SK-Hep1-NIS at 10 and 30 min and at 1, 2, 6, 16, and 23 h after injection of 125I, 99mTc- pertechnetate, or 188Re-perrhenate. In tumor imaging studies, the nude mice were intravenously injected with 188Re-perrhenate and imaged with a gamma-camera equipped with a pinhole collimator at 30 and 60 min after injection. The survival rate (%) was determined by the clonogenic assay after 37 MBq/10 mL (1 mCi/10 mL) 131I and 188Re-perrhenate treatment. RESULTS: SK-Hep1-NIS, stably expressing the NIS gene, accumulated 125I up 150 times higher than that of SK-Hep1. Iodine uptake of SK-Hep1-NIS is completely blocked by perchlorate. NIS gene transfection into SK-Hep1 also resulted in 112- and 87-fold increases of 99mTc-pertechnetate and 188Re-perrhenate uptake, respectively. Iodide efflux from SK-Hep1-NIS was relatively slow, with only 10% released during the initial 5 min, and 60% remained at 25 min. In the biodistribution study using SK-Hep1-NIS-xenographed mice, the tumor uptake of 125I, 188Re-perrhenate, and 99mTc-pertechnetate was 68.0 +/- 15.0, 46.2 +/- 9.1, and 59.6 +/- 16.2 %ID/g (percentage injected dose per gram) at 2 h after injection, respectively. After 188Re-perrhenate injection in SK-Hep1 and SK-Hep1-NIS-xenographed nude mice, whole-body images clearly visualized the SK-Hep1-NIS tumor, whereas the control tumor was not visualized. The survival rate (%) of SK-Hep1-NIS was markedly reduced to 46.3% +/- 10.1% and 28.9% +/- 5.2% after 37 MBq/mL (1 mCi/10 mL) 131I and 188Re-perrhenate treatment compared with the survival rates of the parental cells. These results demonstrated that SK-Hep1-NIS could be selectively killed by the induced 131I and 188Re-perrhenate accumulation through NIS gene expression. CONCLUSION: NIS-based gene therapy using beta-emitting radionuclides has the potential to be used in hepatocellular carcinoma management. PMID- 15347727 TI - Arterial input function measurement without blood sampling using a beta microprobe in rats. AB - The evaluation of every new radiotracer involves pharmacokinetic studies on small animals to determine its biodistribution and local kinetics. To extract relevant biochemical information, time-activity curves for the regions of interest are mathematically modeled on the basis of compartmental models that require knowledge of the time course of the tracer concentration in plasma. Such a time activity curve, usually termed input function, is determined in small animals by repeated blood sampling and subsequent counting in a well counter. The aim of the present work was to propose an alternative to blood sampling in small animals, since this procedure is labor intensive, exposes the staff to radiation, and leads to an important loss of blood, which affects hematologic parameters. METHODS: Monte Carlo simulations were performed to evaluate the feasibility of measuring the arterial input function using a positron-sensitive microprobe placed in the femoral artery of a rat. The simulation results showed that a second probe inserted above the artery was necessary to allow proper subtraction of the background signal arising from tracer accumulation in surrounding tissues. This approach was then validated in vivo in 5 anesthetized rats. In a second set of experiments, on 3 rats, a third probe was used to simultaneously determine 18F FDG accumulation in the striatum. RESULTS: The high temporal resolution of the technique allowed accurate determination of the input function peak after bolus injection of 18F-FDG. Quantitative input functions were obtained after normalization of the arterial time-activity curve for a late blood sample. In the second set of experiments, compartmental modeling was achieved using either the blood samples or the microprobe data as the input function, and similar kinetic constants were found in both cases. CONCLUSION: Although direct quantification proved difficult, the microprobe allowed accurate measurement of arterial input function with a high temporal resolution and no blood loss. The technique, because offering adequate sensitivity and temporal resolution for kinetic measurements of radiotracers in the blood compartment, should facilitate quantitative modeling for radiotracer studies in small animals. PMID- 15347728 TI - Comparison of methods to quantitate 18F-FDG uptake with PET during experimental acute lung injury. AB - PET with 18F-FDG may be useful for quantifying neutrophilic activation. We previously demonstrated that pulmonary neutrophil sequestration could be detected during acute lung injury (ALI), even without migration into the alveolar compartment. Using the influx constant Ki as the method to quantify lung 18F-FDG uptake, we also showed that Ki correlated positively with in vitro assays of 3H deoxyglucose (3H-DG) uptake in cells harvested via bronchoalveolar lavage. In the present study, we have reanalyzed data from that study to determine if simpler nonkinetic methods of quantifying the pulmonary uptake of 18F-FDG could be as powerful as calculating Ki. METHODS: 18F-FDG uptake was quantified as Ki, calculated by 3-compartmental model analysis (used as the gold standard) and Patlak graphical analysis, with and without normalization for initial volume of tracer distribution; the standardized uptake value; and the tissue-to-plasma activity ratio (TPR). RESULTS: Values for Ki, determined either from a 3 compartmental model analysis of the time-activity data or by Patlak graphical analysis, were highly correlated (R2 = 0.97). The correlation was worse if these variables were normalized for the initial volume of tracer distribution. TPR was highly correlated with Ki determined by the compartmental model (R2 = 0.96) and with in vitro measurements of 3H-DG uptake (R2 = 0.63). CONCLUSION: The TPR is a simple and equally effective alternative to dynamic imaging in determining net 18F-FDG uptake during ALI. Normalization of the kinetic data for differences in the initial volume of tracer distribution does not contribute significantly to signal interpretation during ALI. PMID- 15347729 TI - SPECT and PET amino acid tracer influx via system L (h4F2hc-hLAT1) and its transstimulation. AB - System L amino acid transport is increased in various types of cancer. The tracer 123I-2-iodotyrosine (2IT), which is accumulated via system L, could thus serve to allow visualization of cancer in vivo. Here, we studied the transport of 125I-2IT by h4F2hc-hLAT1, the major transporter subserving system L in growing cells, using the Xenopus laevis oocyte expression system. We compared the apparent affinity of 125I-2IT with that of tyrosine, tested the influence of intracellular methionine availability on the influx rate of this substrate, and then compared the transport of 2IT with that of the other tracers-iodo-alpha-methyltyrosine (IMT), fluoroethyltyrosine (FET), and 2-fluorotyrosine (2FT)-by measuring their transstimulating effect on phenylalanine efflux. METHODS: Transport experiments were performed with Xenopus laevis oocytes expressing h4F2hc-hLAT1 (the functional transporter) and oocytes expressing only h4F2hc (negative control). The values obtained for the functional transporter were corrected for endogenous background transport by subtracting the values for the negative controls. RESULTS: The apparent affinity for 125I-2IT and 3H-tyrosine was 29.3 +/- 9.3 micromol/L and 21.2 +/- 4.2 micromol/L, respectively. The influx rate of 125I-2IT was, similarly to that of 3H-phenylalanine, transstimulated by a factor of > or =3 when the oocytes were preinjected with methionine or phenylalanine. The proportion of preinjected 3H-phenylalanine that effluxed within 90 s in the presence of an extracellular 2IT concentration of 0.1 mmol/L was 4.1% +/- 0.5%, compared with 3.3% +/- 0.4% for extracellular IMT, 1.3% +/- 0.3% for FET, 9.3% +/ 0.8% for 2FT, and 9.1% +/- 0.5% for phenylalanine. CONCLUSION: 2IT has a high affinity for h4F2hc-hLAT1, comparable to that of natural tyrosine, and its influx rate is transstimulated by intracellular amino acids. The 2IT influx rate is comparable to that of IMT but lower than that of phenylalanine. In contrast to FET, which is only poorly transported, 2FT displays a high influx rate equal to that of phenylalanine. PMID- 15347730 TI - False-positive findings on myocardial perfusion SPECT. PMID- 15347731 TI - Periareolar injections and hot sentinel nodes. PMID- 15347732 TI - Using [11C]diprenorphine to image opioid receptor occupancy by methadone in opioid addiction: clinical and preclinical studies. AB - Substitute methadone prescribing is one of the main modes of treatment for opioid dependence with established evidence for improved health and social outcomes. However, the pharmacology underpinning the effects of methadone is little studied despite controversies about dosing in relation to outcome. We therefore examined the relationship between methadone dose and occupation of opioid receptors in brain using the positron emission tomography (PET) radioligand [(11)C]diprenorphine in humans and rats. Eight opioid-dependent subjects stable on their substitute methadone (18-90 mg daily) had an [(11)C]diprenorphine PET scan at predicted peak plasma levels of methadone. These were compared with eight healthy controls. No difference in [(11)C]diprenorphine binding was found between the groups, with no relationship between methadone dose and occupancy. Adult male Sprague-Dawley rats that had been given an acute i.v. injection of methadone hydrochloride (0.35, 0.5, 0.7, or 1.0 mg kg(-1)) before [(11)C]diprenorphine showed a dose-dependent increase in biodistribution but no reduction in [(11)C]diprenorphine binding. We suggest that the lack of a dose-dependent relationship between methadone dose, either given chronically in human or acutely in rat, and occupancy of opioid receptor measured with [(11)C]diprenorphine PET is related to efficacy of this opioid agonist at very low levels of opioid receptor occupancy. This has implications for understanding the actions of methadone in comparison with other opioid drugs such as partial agonists and antagonists. PMID- 15347733 TI - Critical role of the atypical {lambda} isoform of protein kinase C (PKC-{lambda}) in oxidant-induced disruption of the microtubule cytoskeleton and barrier function of intestinal epithelium. AB - Oxidant injury to epithelial cells and gut barrier disruption are key factors in the pathogenesis of inflammatory bowel disease. Studying monolayers of intestinal (Caco-2) cells, we reported that oxidants disrupt the cytoskeleton and cause barrier dysfunction (hyperpermeability). Because the lambda isoform of protein kinase C (PKC-lambda), an atypical diacylglycerol-independent isozyme, is abundant in parental (wild type) Caco-2 cells and is translocated to the particulate fractions upon oxidant exposure, we hypothesized that PKC-lambda is critical to oxidative injury to the assembly and architecture of cytoskeleton and the intestinal barrier function. To this end, Caco-2 cells were transfected with an inducible plasmid, a tetracycline-responsive system, to create novel clones stably overexpressing native PKC-lambda. Other cells were transfected with a dominant-negative plasmid to stably inhibit the activity of native PKC-lambda. Cells were exposed to oxidant (H(2)O(2)) +/- modulators. Parental Caco-2 cells were treated similarly. We then monitored barrier function (fluorescein sulfonic acid clearance), microtubule cytoskeletal stability (confocal microscopy, immunoblotting), subcellular distribution of PKC-lambda (immunofluorescence, immunoblotting, immunoprecipitation), and PKC-lambda isoform activity (in vitro kinase assay). Monolayers were also processed to assess alterations in tubulin assembly, polymerized tubulin (S2, an index of cytoskeletal integrity), and monomeric tubulin (S1, an index of cytoskeletal disassembly) (polyacrylamide gel electrophoresis fractionation and immunoblotting. In parental cells, oxidant caused: 1) translocation of PKC-lambda from the cytosol to the particulate (membrane + cytoskeletal) fractions, 2) activation of native PKC-lambda, 3) tubulin pool instability (increased monomeric S1 and decreased polymerized S2), 4) disruption of cytoskeletal architecture, and 5) barrier dysfunction (hyperpermeability). In transfected clones, overexpression of the atypical (74 kDa) PKC-lambda isoform by itself ( approximately 3.2-fold increase) led to oxidant-like disruptive effects, including cytoskeletal and barrier hyperpermeability. Overexpressed PKC-lambda was mostly found in particulate cell fractions (with a smaller cytosolic distribution) indicating its activation. Disruption by PKC-lambda overexpression was also potentiated by oxidant challenge. Stable inactivation of endogenous PKC-lambda ( approximately 99.6%) by a dominant-negative protected against all measures of oxidant-induced disruption. We conclude that: 1) oxidant induces disruption of epithelial barrier integrity by disassembling the cytoskeleton, in large part, through the activation of PKC lambda isoform; and 2) activation of PKC-lambda by itself appears to be sufficient for disruption of cellular cytoskeleton and monolayer barrier permeability. The unique ability to mediate an oxidant-like injury and cytoskeletal depolymerization and instability is a novel mechanism not previously attributed to the atypical subfamily of PKC isoforms. PMID- 15347734 TI - A selective androgen receptor modulator for hormonal male contraception. AB - The recent discovery of nonsteroidal selective androgen receptor modulators (SARMs) provides a promising alternative for testosterone replacement therapies, including hormonal male contraception. The identification of an orally bioavailable SARM with the ability to mimic the central and peripheral androgenic and anabolic effects of testosterone would represent an important step toward the "male pill". We characterized the in vitro and in vivo pharmacologic activity of (S)-3-(4-chloro-3-fluorophenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3 trifluoromethylphenyl)propionamide (C-6), a novel SARM developed in our laboratories. C-6 was identified as an androgen receptor (AR) agonist with high AR binding affinity (K(i) = 4.9 nM). C-6 showed tissue-selective pharmacologic activity with higher anabolic activity than androgenic activity in male rats. The doses required to maintain the weight of the prostate, seminal vesicles, and levator ani muscle to half the size of the maximum effects (i.e., ED(50)) were 0.78 +/- 0.06, 0.88 +/- 0.1, and 0.17 +/- 0.04 mg/day, respectively. As opposed to other SARMs, gonadotropin levels in C-6-treated groups were significantly lower than control values. C-6 also significantly decreased serum testosterone concentration in intact rats after 2 weeks of treatment. Marked suppression of spermatogenesis was observed after 10 weeks of treatment with C-6 in intact male rats. Pharmacokinetic studies of C-6 in male rats revealed that C-6 was well absorbed after oral administration (bioavailability 76%), with a long (6.3 h) half-life at a dose of 10 mg/kg. These studies show that C-6 mimicked the in vivo pharmacologic and endocrine effects of testosterone while maintaining the oral bioavailability and tissue-selective actions of nonsteroidal SARMs. PMID- 15347735 TI - Treatment with {alpha}-phenyl-N-tert-butylnitrone, a free radical-trapping agent, abrogates inflammatory cytokine gene expression during alloimmune activation in rat cardiac allografts. AB - Spin-trapping nitrones such as alpha-phenyl-N-tert-butylnitrone (PBN) have traditionally been used to trap and stabilize free radicals for detection by electron paramagnetic resonance (EPR) spectroscopy. Unlike classical antioxidants, these agents have never been evaluated therapeutically in allograft transplantation. In the present study, we examined potential mechanisms of action of treatment with PBN in a rat model of acute cardiac allograft transplantation. Graft rejection was determined by histological examination and graft function determined by in situ sonomicrometry. DNA binding for nuclear factor (NF)-kappaB and activator protein (AP-1) were determined by gel shift assays. Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed for inducible nitric-oxide synthase (iNOS) and inflammatory cytokines. Histological rejection scores were elevated in untreated allografts and decreased by treatment with PBN. In situ sonomicrometry revealed decreased heart rate and distended end diastolic and end systolic segment lengths with rejection. Although PBN did not alter heart rate, it did normalize the distention of both diastolic and systolic cardiac dimension. EPR spectroscopy revealed nitrosylation of myocardial heme protein in untreated allografts that was decreased by treatment with PBN. PBN also decreased iNOS protein and iNOS mRNA. RT-PCR analysis revealed enhanced cytokine gene expression for interferon-gamma, interleukin-6, and interleukin-10 in untreated allografts. Expression for these genes was potently inhibited or abolished in recipients treated with PBN. PBN treatment also decreased DNA binding of transcription factors, NF-kappaB and AP-1. Thus, PBN retains significant anti-inflammatory properties through its action to down regulate cytokine gene expression that contribute to protection against acute alloimmune activation in cardiac allografts. PMID- 15347736 TI - AZF and DAZ gene copy-specific deletion analysis in maturation arrest and Sertoli cell-only syndrome. AB - Deletions of the AZFc region in Yq11.2, which include the DAZ gene family, are responsible for most cases of male infertility and were associated with severe oligozoospermia and also with a variable testicular pathology. To uncover the functional contribution of DAZ to human spermatogenesis, a DAZ gene copy-specific deletion analysis was previously established and showed that DAZ1/DAZ2 deletions associate with oligozoospermia. In this study we applied the same screening method to 50 control fertile males and 91 non-obstructive azoospermic males, 39 with Sertoli cell-only syndrome (SCOS) and 52 with meiotic arrest (MA). Samples were also screened with 24 sequence-tagged sites to the different AZF regions, including 114 control fertile males. After biopsy (testicular sperm extraction, TESE), residual spermiogenesis was found in 57.7% MA and 30.8% SCOS cases (incomplete syndromes). DAZ1/DAZ2 deletions were associated with the testicular phenotype of residual spermiogenesis as they were only found in two patients (8%) with incomplete MA. Differences between incomplete (23.3%) and complete (4.5%) MA cases regarding AZFc and DAZ1/DAZ2 deletion frequencies, and between incomplete (58.3%) and complete (11.1%) SCOS cases for AZFc deletions, suggest that incomplete syndromes might represent an aggravation of the oligozoospermic phenotype. As successful TESE was achieved in 87.5% of MA cases with AZFc and DAZ1/DAZ2 deletions and in 58.3% of SCOS cases with AZFc deletions, the present results also suggest that these molecular markers might be used for the establishment of a prognosis before TESE. PMID- 15347737 TI - Silencing lamin A/C in human endometrial stromal cells: a model to investigate endometrial gene function and regulation. AB - Silencing of a target mRNA by small interfering RNA (siRNA) has emerged as a new and powerful tool to study gene function, and post-transcriptional gene silencing can now be accomplished with 21-23 nucleotide RNA that mediate sequence-specific mRNA degradation. In the current study we employed lamin A/C siRNA to silence lamin A/C expression in cultured human endometrial stromal cells and investigated downstream cellular markers for proof of concept. Human endometrial stromal cells from three subjects were transfected with lamin A/C siRNA or non-silencing fluorescein-labelled siRNA, and flow cytometric analysis revealed 95-98% transfection efficiency after 6 h of treatment. RT-PCR and quantitative RT-PCR were used to measure mRNA degradation of lamin A/C, and 75-88% silencing was observed 48 h post-transfection. Western blotting and immunocytochemistry confirmed corresponding decrease in lamin A/C protein within 48 h of gene silencing. The downstream effect of lamin A/C silencing was investigated by immunocytochemical analysis of the cellular localization of the protein, emerin, an important component of the nuclear lamina and known to be regulated by lamin expression. Marked displacement of emerin from the nuclear lamina to the cytoplasm was observed when lamin A/C was silenced in human endometrial stromal cells, confirming functional silencing of lamin A/C resulting in a nuclear lamina assembly defect. Silencing target mRNA by siRNA in human endometrial stromal cells can be more broadly applied to investigate the function and regulation of other genes in this cell type, and the methodology and data presented herein strongly support the more widespread use of this powerful tool in endometrial biology research. PMID- 15347738 TI - Genome update: promoter profiles. PMID- 15347739 TI - Sensing and responding to diverse extracellular signals? Analysis of the sensor kinases and response regulators of Streptomyces coelicolor A3(2). AB - Streptomyces coelicolor is a Gram-positive soil bacterium that undergoes a complex developmental life cycle. The genome sequence of this organism was recently completed and has revealed the presence of over 60 sigma factors and a multitude of other transcriptional regulators, with a significant number of these being putative two-component signal transduction proteins. The authors have used the criteria established by Hoch and co-workers (Fabret et al., 1999, J Bacteriol 181, 1975-1983) to identify sensor kinase and response regulator genes encoded within the S. coelicolor genome. This analysis has revealed the presence of 84 sensor kinase genes, 67 of which lie adjacent to genes encoding response regulators. This strongly suggests that these paired genes encode two-component systems. In addition there are 13 orphan response regulators encoded in the genome, several of which have already been characterized and are implicated in development and antibiotic production, and 17 unpaired and as yet uncharacterized sensor kinases. This article attempts to infer useful information from sequence analysis and reviews what is currently known about the two-component systems, unpaired sensor kinases and orphan response regulators of S. coelicolor from both published reports and the authors' own unpublished data. PMID- 15347740 TI - Polymorphonuclear leukocytes (PMNs) induce protective Th1-type cytokine epithelial responses in an in vitro model of oral candidosis. AB - The immune response and the anticandidal activity of keratinocytes and polymorphonuclear leukocytes (PMNs) play a key role in host defence against localized Candida albicans infection. An established model of oral candidosis based on reconstituted human oral epithelium (RHE) was supplemented with PMNs to study the effect of these immune cells during experimental oral candidosis. Infection of RHE with C. albicans induced a strong expression of the chemokine interleukin-8 (IL-8) and the cytokine granulocyte-macrophages colony-stimulating factor (GM-CSF), and a moderate stimulation of interleukin-1alpha (IL-1alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interferon gamma (IFN-gamma) and tumour necrosis factor alpha (TNF-alpha) by keratinocytes. This immune response was associated with chemoattraction of PMNs to the site of infection, whereas uninfected RHE failed to induce cytokine expression or to attract PMNs. Growth of the pathogen and tissue damage of C. albicans-infected RHE were significantly reduced when PMNs were applied to the apical epithelial surface or when PMNs migrated through a perforated basal polycarbonate filter of the model. Notably, protection against epithelial tissue damage was also observed when PMNs were placed on the basal side of non-perforated filters, which prevented PMN migration into the RHE. Addition of PMNs enhanced a Th1-type immune response (IFN gamma, TNF-alpha), down-regulated the expression of the Th2-type cytokine interleukin-10 (IL-10), and was associated with protection against Candida induced tissue damage. This PMN-supplemented model of oral candidosis mimics the in vivo situation, and provides a promising tool for studying the immunological interactions between keratinocytes and C. albicans, as well as the influence of PMNs on C. albicans pathogenesis. PMID- 15347741 TI - Dynamic localization of membrane proteins in Bacillus subtilis. AB - The subcellular localization of membrane proteins in Bacillus subtilis was examined by using fluorescent protein fusions. ATP synthase and succinate dehydrogenase were found to localize within discrete domains on the membrane rather than being homogeneously distributed around the cell periphery as expected. Dual labelling of cells indicated partial colocalization of ATP synthase and succinate dehydrogenase. Further analysis using an ectopically expressed phage protein gave the same localization patterns as ATP synthase and succinate dehydrogenase, implying that membrane proteins are restricted to domains within the membrane. 3D reconstruction of images of the localization of ATP synthase showed that domains were not regular and there was no bias for localization to cell poles or any other positions. Further analysis revealed that this localization was highly dynamic, but random, implying that integral membrane proteins are free to diffuse two-dimensionally around the cytoplasmic membrane. PMID- 15347742 TI - Calnexin, calreticulin and cytoskeleton-associated proteins modulate uptake and growth of Legionella pneumophila in Dictyostelium discoideum. AB - The haploid amoeba Dictyostelium discoideum is a versatile host system for studying cellular aspects of Legionella pathogenicity. Previous studies have shown that the internalization of L. pneumophila leads to an endoplasmic reticulum (ER)-derived organelle that supports intracellular replication of the bacteria. In this study a roadmap of host-cell factors involved in this process was developed. Phagocytosis assays with specific cellular inhibitors and the effects of well defined host-cell mutants revealed that cytoplasmic calcium levels, cytoskeleton-associated proteins and the calcium-binding proteins of the ER, calreticulin and calnexin, specifically influence the uptake and intracellular growth of L. pneumophila. Confocal microscopic time series with green fluorescent protein (GFP)-tagged calnexin and calreticulin demonstrated the accumulation of both proteins in the phagocytic cup of L. pneumophila-infected host cells. In contrast to the control experiment with Escherichia coli containing phagosomes, both proteins decorated the replicative vacuole of L. pneumophila during the entire growth phase of the bacteria. The cumulative effects of cytosolic calcium levels, the spatial distribution of calnexin and calreticulin, and the defective invasion and replication of L. pneumophila in calnexin- and calreticulin-minus cells suggest that these factors are part of a regulatory system that leads to the specific vacuole of L. pneumophila. PMID- 15347743 TI - Balamuthia mandrillaris from soil samples. AB - Balamuthia mandrillaris amoebas are recognized as a causative agent of granulomatous amoebic encephalitis, a disease that is usually fatal. They were first recognized when isolated from the brain of a mandrill baboon that died in the San Diego Zoo Wild Life Animal Park. Subsequently, the amoebas have been found in a variety of animals, including humans (young and old, immunocompromised and immunocompetent persons), in countries around the world. Until recently, the amoebas had not been recovered from the environment and their free-living status was in question. The recovery of a Balamuthia amoeba from a soil sample taken from a plant at the home of a child from California, USA, who died of Balamuthia amoebic encephalitis, was reported previously. In a continued investigation, a second amoeba was isolated from soil that was obtained from an outdoor potted plant in a spatially unrelated location. A comparison of these two environmental amoebas that were isolated from different soils with the amoeba that was obtained from the child's clinical specimen is reported here. Included are the isolation procedure for the amoebas, their growth requirements, their immunological response to anti-Balamuthia serum, their sensitivity to a selection of antimicrobials and sequence analysis of their 16S rRNA gene. The evidence is consistent that the amoebas isolated from both soil samples and the clinical isolate obtained from the Californian child are B. mandrillaris. PMID- 15347744 TI - Ciliostasis is a key early event during colonization of canine tracheal tissue by Bordetella bronchiseptica. AB - The primary site of infection for Bordetella bronchiseptica, Bordetella pertussis and Bordetella parapertussis is the ciliated respiratory epithelium. Previous studies have implicated adherence of bacteria to cilia, induction of mucus production, induction of ciliostasis and damage to the ciliated epithelium in Bordetella pathogenesis. This paper describes the use of an air-interface organ culture system using canine tracheal tissue infected with B. bronchiseptica to assess the temporal relationship between these pathologies. Ciliostasis occurs very early during the host tissue-pathogen interaction, before mucus production and obvious signs of epithelial damage occur. A B. bronchiseptica bvg mutant does not colonize the organ culture model, induce ciliostasis or cause damage to the epithelial cell layer, but it does induce similar amounts of mucus release as does infection by wild-type bacteria. The authors propose that ciliostasis is a key early event during the B. bronchiseptica-host tissue interaction that abrogates the muco-ciliary defences of the host tissue, renders it susceptible to colonization by the bacteria and allows subsequent damage to the epithelium. The organ culture model described offers a physiologically relevant tool with which to characterize the molecular basis for interactions between Bordetella and its primary site of infection, the ciliated respiratory epithelium. PMID- 15347745 TI - Transcription of Proteus mirabilis flaAB. AB - Proteus mirabilis, a Gram-negative urinary tract pathogen, has two highly homologous, tandemly arranged flagellin-encoding genes, flaA and flaB. flaA is transcribed from a sigma(28) promoter, while flaB is a silent allele. Previous studies have demonstrated the presence of a family of hybrid flagellin genes, referred to as flaAB. These genes are composed of the 5' end of flaA and the 3' end of flaB, and are produced through excision of the intervening DNA between the two genes. Although the existence of flaAB DNA has been documented, it was not known if transcription of flaAB occurs in wild-type P. mirabilis. In this study, proof of flaAB transcription was obtained from a combination of RNA dot-blots and RT-PCR assays using specific primers and probes for flaAB and flaA. The RNA data were further supported by the demonstration of phenotypic switching of the locus using a FlaAB-detector strain. The results show that flaAB mRNA is transcribed and is 1/64 as abundant as flaA in the population of wild-type cells, suggesting that flaAB constitutes 1.0-1.5 % of the total flagellin message. Nucleotide sequence analysis of flaAB products produced by RT-PCR from the wild-type confirms previous reports of a variable fusion site between flaA and flaB resulting in a hybrid flagellin transcript. These data support the hypothesis that the production of FlaAB is integral to the physiology of P. mirabilis. PMID- 15347746 TI - New roles for CDC25 in growth control, galactose regulation and cellular differentiation in Saccharomyces cerevisiae. AB - Living organisms display large differences in stress resistance throughout their life cycles. To study the coordinated regulation of development and stress responses in exponentially growing yeast, mutants that displayed elevated heat shock resistance at this stage were screened for. Here, two new mutant alleles of CDC25 in Saccharomyces cerevisiae, cdc25-21 and cdc25-22, are described. During exponential growth in glucose at 25 degrees C, these mutants are resistant to heat, oxidative, osmotic and ionic shock, accumulate stress-protein transcripts, show slow growth rates, thick cell walls and glycogen hyperaccumulation and lack cAMP signalling in response to glucose. Genetic and cellular analyses revealed that the stationary-phase phenotypes of cdc25-21 and cdc25-22 mutants are not due to entrance to a G(0) state during exponential growth, but are the result of a prolonged G(1) phase. It was found that, in the W303 background, CDC25 is dispensable for growth in glucose media. However, CDC25 is essential for growth in galactose, in non-fermentable carbon sources and under continuous incubation at 38 degrees C. In conclusion, the function of the catalytic, C-terminal domain of Cdc25p is not only important for fermentative growth, but also for growth in non-fermentable carbon sources and to trigger galactose derepression. PMID- 15347747 TI - Characterization of the Aspergillus parasiticus delta12-desaturase gene: a role for lipid metabolism in the Aspergillus-seed interaction. AB - In the mycotoxigenic oilseed pathogens Aspergillus flavus and Aspergillus parasiticus and the model filamentous fungus Aspergillus nidulans, unsaturated fatty acids and their derivatives act as important developmental signals that affect asexual conidiospore, sexual ascospore and/or sclerotial development. To dissect the relationship between lipid metabolism and fungal development, an A. parasiticus Delta(12)-desaturase mutant that was unable to convert oleic acid to linoleic acid and was thus impaired in polyunsaturated fatty acid biosynthesis was generated. The Delta(12)-desaturase mutant demonstrates delayed spore germination, a twofold reduction in growth, a reduced level of conidiation and complete loss of sclerotial development, compared to the wild-type. Host colonization is impaired, as reflected by a decrease in conidial production on live peanut and corn seed by the mutant compared to the wild-type. Similarly, the previously isolated A. nidulans Delta(12)-desaturase mutant has reduced colonization capabilities compared to the wild-type. Therefore, desaturation mutants display a key requisite that affords a genetic solution to oilseed crop contamination by mycotoxigenic Aspergillus species: a reduction in the production of conidia, the infectious particle of the pathogenic aspergilli. PMID- 15347748 TI - Functional and phylogenetic analysis of a plant-inducible oligoribonuclease (orn) gene from an indigenous Pseudomonas plasmid. AB - Application of a promoter-trapping strategy to identify plant-inducible genes carried on an indigenous Pseudomonas plasmid, pQBR103, revealed the presence of a putative oligoribonuclease (orn) gene that encodes a highly conserved 3' to 5' exoribonuclease specific for small oligoribonucleotides. The deduced amino acid sequence of the plasmid-derived orn (orn(pl)) showed three conserved motifs characteristic of Orn from both prokaryotes and eukaryotes. Deletion of orn(pl) generated no observable phenotype, but inactivation of the chromosomal copy caused slow growth in Pseudomonas putida KT2440. This defect was fully restored by complementation with orn from Escherichia coli (orn(E.coli)). Plasmid-derived orn(pl) was capable of partially complementing the P. putida orn mutant, demonstrating functionality of orn(pl). Phylogenetic analysis showed that plasmid encoded Orn was distinct from Orn encoded by the chromosome of proteobacteria. A survey of orn(pl) from related Pseudomonas plasmids showed a sporadic distribution but no sequence diversity. These data suggest that the orn(pl) was acquired by pQBR103 in a single gene-transfer event: the donor is unknown, but is unlikely to be a member of the Proteobacteria. PMID- 15347749 TI - Use of community genome arrays (CGAs) to assess the effects of Acacia angustissima on rumen ecology. AB - This research developed a community genome array (CGA) to assess the effects of Acacia angustissima on rumen microbiology. A. angustissima produces non-protein amino acids as well as tannins, which may be toxic to animals, and CGA was used to assess the effects of this plant on the ecology of the rumen. CGAs were developed using a 7.5 cmx2.5 cm nylon membrane format that included up to 96 bacterial genomes. It was possible to separately hybridize large numbers of membranes at once using this mini-membrane format. Pair-wise cross-hybridization experiments were conducted to determine the degree of cross-hybridization between strains; cross-hybridization occurred between strains of the same species, but little cross-reactivity was observed among different species. CGAs were successfully used to survey the microbial communities of animals consuming an A. angustissima containing diet but quantification was not precise. To properly quantify and validate the CGA, Fibrobacter and Ruminococcus populations were independently assessed using 16S rDNA probes to extracted rRNA. The CGA detected an increase in these populations as acacia increased in the diet, which was confirmed by rRNA analysis. There was a great deal of variation among strains of the same species in how they responded to A. angustissima. However, in general Selenomonas strains tended to be resistant to the tannins in the acacia while Butyrivibrio fibrisolvens was sensitive. On the other hand some species, like streptococci, varied. Streptococcus bovis-like strains were sensitive to an increase in acacia in the diet while Streptococcus gallolyticus-like strains were resistant. Strep. gallolyticus has independently been shown to be resistant to tannins. It is concluded that there is significant variation in tannin resistance between strains of the same species. This implies that there are specific molecular mechanisms at play that are independent of the phylogenetic position of the organism. PMID- 15347750 TI - Roles and regulation of the glutamate racemase isogenes, racE and yrpC, in Bacillus subtilis. AB - Many bacteria, including Escherichia coli, have a unique gene that encodes glutamate racemase. This enzyme catalyses the formation of d-glutamate, which is necessary for cell wall peptidoglycan synthesis. However, Bacillus subtilis has two glutamate racemase genes, named racE and yrpC. Since racE appears to be indispensable for growth in rich medium, the role of yrpC in d-amino acid synthesis is vague. Experiments with racE- and yrpC-knockout mutants confirmed that racE is essential for growth in rich medium but showed that this gene was dispensable for growth in minimal medium, where yrpC executes the anaplerotic role of racE. LacZ fusion assays demonstrated that racE was expressed in both types of media but yrpC was expressed only in minimal medium, which accounted for the absence of yrpC function in rich medium. Neither racE nor yrpC was required for B. subtilis cells to synthesize poly-gamma-dl-glutamate (gamma-PGA), a capsule polypeptide of d- and l-glutamate linked through a gamma-carboxylamide bond. Wild-type cells degraded the capsule during the late stationary phase without accumulating the degradation products, d-glutamate and l-glutamate, in the medium. In contrast, racE or yrpC mutant cells accumulated significant amounts of d- but not l-glutamate. Exogenous d-glutamate utilization was somewhat defective in the mutants and the double mutation of race and yrpc severely impaired d-amino acid utilization. Thus, both racemase genes appear necessary to complete the catabolism of exogenous d-glutamate generated from gamma-PGA. PMID- 15347751 TI - Metabolic properties of Eubacterium pyruvativorans, a ruminal 'hyper-ammonia producing' anaerobe with metabolic properties analogous to those of Clostridium kluyveri. AB - Eubacterium pyruvativorans I-6(T) is a non-saccharolytic, amino-acid-fermenting anaerobe from the rumen, isolated by its ability to grow on pancreatic casein hydrolysate (PCH) as sole C source. This study investigated its metabolic properties and its likely ecological niche. Additional growth was supported by pyruvate, vinyl acetate, and, to a lesser extent, lactate and crotonate, and also by a mixture of amino acids (alanine, glycine, serine and threonine) predicted to be catabolized to pyruvate. No single amino acid supported growth, and peptides were required for growth on amino acids. Alanine, followed by leucine, serine and proline, were used most extensively during growth, but only alanine and asparate were extensively modified before incorporation. Growth on PCH, but not on pyruvate, was increased by the addition of acetate, propionate and butyrate. l Lactate was fermented incompletely, mainly to acetate, but no lactate-C was incorporated. Propionate and butyrate were utilized during growth, forming valerate and caproate, respectively. Labelling experiments suggested a metabolic pattern where two C atoms of butyrate, valerate and caproate were derived from amino acids, with the others being formed from acetate, propionate and butyrate. The metabolic strategy of E. pyruvativorans therefore resembles that of Clostridium kluyveri, which ferments ethanol only when the reaction is coupled to acetate, propionate or butyrate utilization. The fermentative niche of E. pyruvativorans appears to be to scavenge amino acids, lactate and possibly other metabolites in order to generate ATP via acetate formation, using volatile fatty acid elongation with C(2) units derived from other substrates to dispose of reducing equivalents. PMID- 15347752 TI - Evidence for a copper-dependent iron transport system in the marine, magnetotactic bacterium strain MV-1. AB - Cells of the magnetotactic marine vibrio, strain MV-1, produce magnetite containing magnetosomes when grown anaerobically or microaerobically. Stable, spontaneous, non-magnetotactic mutants were regularly observed when cells of MV-1 were cultured on solid media incubated under anaerobic or microaerobic conditions. Randomly amplified polymorphic DNA analysis showed that these mutants are not all genetically identical. Cellular iron content of one non-magnetotactic mutant strain, designated MV-1nm1, grown anaerobically, was approximately 20- to 80-fold less than the iron content of wild-type (wt) MV-1 for the same iron concentrations, indicating that MV-1nm1 is deficient in some form of iron uptake. Comparative protein profiles of the two strains showed that MV-1nm1 did not produce several proteins produced by wt MV-1. To understand the potential roles of these proteins in iron transport better, one of these proteins was purified and characterized. This protein, a homodimer with an apparent subunit mass of about 19 kDa, was an iron-regulated, periplasmic protein (p19). Two potential 'copper-handling' motifs (MXM/MX(2)M) are present in the amino acid sequence of p19, and the native protein binds copper in a 1 : 1 ratio. The structural gene for p19, chpA (copper handling protein) and two other putative genes upstream of chpA were cloned and sequenced. These putative genes encode a protein similar to the iron permease, Ftr1, from the yeast Saccharomyces cerevisiae, and a ferredoxin-like protein of unknown function. A periplasmic, copper-containing, iron(II) oxidase was also purified from wt MV-1 and MV-1nm1. This enzyme, like p19, was regulated by media iron concentration and contained four copper atoms per molecule of enzyme. It is hypothesized that ChpA, the iron permease and the iron(II) oxidase might have analogous functions for the three components of the S. cerevisiae copper-dependent high-affinity iron uptake system (Ctr1, Ftr1 and Fet3, respectively), and that strain MV-1 may have a similar iron uptake system. However, iron(II) oxidase purified from both wt MV-1 and MV-1nm1 displayed comparable iron oxidase activities using O(2) as the electron acceptor, indicating that ChpA does not supply the multi-copper iron(II) oxidase with copper. PMID- 15347753 TI - Hypermutable Haemophilus influenzae with mutations in mutS are found in cystic fibrosis sputum. AB - Hypermutable bacterial pathogens exist at surprisingly high prevalence and benefit bacterial populations by promoting adaptation to selective environments, including resistance to antibiotics. Five hundred Haemophilus influenzae isolates were screened for an increased frequency of mutation to resistance to rifampicin, nalidixic acid and spectinomycin: of the 14 hypermutable isolates identified, 12 were isolated from cystic fibrosis (CF) sputum. Analysis by enterobacterial repetitive intergenic consensus (ERIC)-PCR and ribotyping identified eight distinct genetic fingerprints. The hypermutable phenotype of seven of the eight unique isolates was associated with polymorphisms in conserved sites of mutS. Four of the mutant mutS alleles were cloned and failed to complement the mutator phenotype of a mutS : : TSTE mutant of H. influenzae strain Rd KW20. Antibiotic susceptibility testing of the hypermutators identified one beta-lactamase negative ampicillin-resistant (BLNAR) isolate with two isolates producing beta lactamase. Six isolates from the same patient with CF, with the same genetic fingerprint, were clonal by multilocus sequence typing (MLST). In this clone, there was an evolution to higher MIC values for the antibiotics administered to the patient during the period in which the strains were isolated. Hypermutable H. influenzae with mutations in mutS are prevalent, particularly in the CF lung environment, and may be selected for and maintained by antibiotic pressure. PMID- 15347754 TI - Diversity of stx2 converting bacteriophages induced from Shiga-toxin-producing Escherichia coli strains isolated from cattle. AB - The presence of bacteriophages encoding Shiga toxin 2 (stx(2) phages) was analysed in 168 strains of Shiga-toxin-producing Escherichia coli (STEC) isolated from cattle. Following mitomycin C induction, strains carrying stx(2) phages were screened by plaque blot and hybridization with an stx(2)A-probe. In the stx(2) phage-carrying strains, the amounts of phage production, phage DNA extracted and Stx(2) produced after induction were assessed. The induced stx(2) phages were characterized morphologically and genetically. Assays to obtain lysogens from different strains were also carried out and phages induced from the lysogens were compared with those induced from the STEC isolates. Results indicated that 18 % of the strains carried an inducible stx(2) phage. Most of them showed a direct relationship between phage induction and toxin production. Each strain carried only one inducible stx(2) phage, although a few strains had two copies of the stx(2) in the chromosome. The stx(2) phages showed diverse morphology and a wide variability in their genome. Assays to obtain lysogens showed that not all the phages were transduced with the same frequency and only six lysogens were obtained. Phages in the lysogens were the same as those induced from their respective initial STEC host strains, although the induction and relative toxin production of the lysogens varied. Most phages carried the stx(2) gene, while a few carried stx(2) variants. Infectivity of the phages depended on the different hosts, although O157 : H7 was preferentially infected by phages induced from O157 strains. The results show that inducible stx(2) phages are common among STEC of animal origin and that they may enhance the spread of stx(2). PMID- 15347755 TI - Genomic subtraction for the identification of putative new virulence factors of an avian pathogenic Escherichia coli strain of O2 serogroup. AB - To identify putative new virulence factors of avian pathogenic Escherichia coli (APEC) strains, a genomic subtraction was performed between the APEC strain MT512 and the non-pathogenic E. coli strain of avian origin EC79. Seventeen DNA fragments were cloned that were specific for the APEC strain. Among them, nine were identified that were more frequent among pathogenic than non-pathogenic isolates in a collection of 67 avian E. coli. Chromosome or plasmid location, and the nucleotide sequence of these nine fragments were characterized. Four fragments were plasmid-located. The nucleotide sequence of two of them exhibited identity with the sequence of the RepF1B replicon of E. coli plasmids, and the amino-acid deduced sequences from the two other fragments exhibited similarity to the products of genes sitA of Salmonella Typhimurium and iroD of E. coli, which are involved in iron metabolism. Of the five chromosome-located fragments, three were predicted to encode parts of proteins that were significantly homologous to previously described proteins: TktA (transketolase) of Haemophilus influenzae, a FruA (fructokinase) homologue of Listeria innocua and Gp2 (large terminal subunit) of phage 21. The putative products of the two other chromosome-located fragments were homologous to proteins with unknown functions: Z0255 of E. coli strain EDL933 (EHEC) and RatA of Salmonella Typhimurium strain LT2. Both these chromosomal fragments, whose presence is correlated with serogroups O1 and O2 and to the virulence of APEC strains belonging to these serogroups, are good candidates for being part of novel virulence determinants of APEC. Moreover, several fragments were shown to be located close to tRNA selC, asnT or thrW, which suggests they could be part of pathogenicity islands. Six fragments that were shown to be part of whole ORFs present in the APEC strain MT 512 were also present in extra-intestinal pathogenic E. coli (ExPEC) strains of human and animal origin. Thus, the putative novel virulence factors identified in this study could be shared by ExPEC strains of different origins. PMID- 15347756 TI - A differential effect of sigmaS on the expression of the PHO regulon genes of Escherichia coli. AB - The RNA polymerase core associated with sigma(S) transcribes many genes related to stress or to the stationary phase. When cells enter a phase of phosphate starvation, the transcription of several genes and operons, collectively known as the PHO regulon, is strongly induced. The promoters of the PHO genes hitherto analysed are recognized by sigma(D)-associated RNA polymerase. A mutation in the gene that encodes sigma(S), rpoS, significantly increases the level of alkaline phosphatase activity and the overproduction of sigma(S) inhibits it. Other PHO genes such as phoE and ugpB are likewise affected by sigma(S). In contrast, pstS, which encodes a periplasmic phosphate-binding protein and is a negative regulator of PHO, is stimulated by sigma(S). The effect of sigma(S) on the PHO genes is at the transcriptional level. It is shown that a cytosine residue at position -13 is important for the positive effect of sigma(S) on pst. The interpretation of these observations is based on the competition between sigma(S) and sigma(D) for the binding to the core RNA polymerase. PMID- 15347757 TI - Functional diversity of three different DsbA proteins from Neisseria meningitidis. AB - The genome of Neisseria meningitidis serogroup B strain MC58 contains three genes - nmb0278, nmb0294 and nmb0407 - encoding putative homologues of DsbA, a periplasmic thiol disulphide oxidoreductase protein-folding catalyst of the Dsb protein family. DsbA assists the folding of periplasmic and membrane proteins in diverse organisms. While all three cloned genes complemented the DTT sensitivity of dsbA-null Escherichia coli, they showed different activities in folding specific target proteins in this background. NMB0278 protein was the most active in complementing defects in motility and alkaline phosphatase activity, while NMB0294 was the most active in folding periplasmic MalF. NMB0407 showed the weakest activity in all assays. It is extremely unusual for organisms to contain more than one chromosomal dsbA. Among the members of the genus Neisseria, only the meningococcus carries all three of these genes. Strains of Neisseria gonorrhoeae, Neisseria lactamica, Neisseria cinerea and Neisseria polysaccharea contained only homologues of nmb0278 and nmb0407, while Neisseria flava, Neisseria subflava and Neisseria flavescens carried only nmb0294. It is speculated that the versatility of the meningococcus in surviving in different colonizing and invasive disease settings may be derived in part from an enhanced potential to deploy outer-membrane proteins, a consequence of carrying an extended repertoire of protein-folding catalysts. PMID- 15347758 TI - Involvement of genes of genome maintenance in the regulation of phase variation frequencies in Neisseria meningitidis. AB - In Neisseria meningitidis, the reversible expression of surface antigens, i.e. phase variation, results from changes within repeated simple sequence motifs located in coding or promoter regions of the genes involved in their biosynthesis. The mutation rates of these simple sequences, which have a major influence on the generation of phenotypic diversity, can affect the fitness of the population. The aim of the present study was to investigate the involvement of genetic factors involved (mutS and dam) and not yet analysed (drg and dinB) in the regulation of phase variation frequencies of genes associated with a variety of repeat tracts. The frequency of frameshifts occurring in the polycytidine (polyC) tracts associated with siaD, spr and lgtG and in the tetranucleotide (TAAA) repeat tract associated with nadA was determined by colony immunoblotting or using the lacZ gene as a reporter. Inactivation of mutS increased the frequency of phase variation of genes presenting homopolymeric tracts of diverse length. Overexpression of dinB enhanced the instability of the homopolymeric tract associated with siaD. Investigation of the dam locus in a population of genetically distinct N. meningitidis strains revealed that 27 % of strains associated with invasive disease contained the dam gene. In all strains where a Dam function was absent, the drg gene had been inserted into the dam locus. Disruption of dam and drg in strains representative of each genotype, i.e. dam(+)/drg and dam/drg(+), did not modify phase variation frequencies. In contrast to the effects of certain genes on homopolymeric tracts, none of the genetic factors investigated affected the stability of tetranucleotide repeat tracts. PMID- 15347759 TI - Cryptococcus neoformans methionine synthase: expression analysis and requirement for virulence. AB - This paper describes (i) the expression profile of the methionine synthase gene (MET6) in the human pathogenic fungus Cryptococcus neoformans and (ii) the phenotypes of a C. neoformans met6 mutant. In contrast to the MET3 gene, which showed no significant change in expression in any environmental condition tested, the MET6 gene showed a substantial induction in response to methionine and a dramatic transcriptional induction in response to homocysteine. Like a met3 mutant, the met6 mutant was a methionine auxotroph. However, relative to a met3 mutant, the met6 mutant grew very slowly and was less heat-shock resistant. In contrast to a met3 mutant, the met6 mutant lost viability when starved of methionine, and it was deficient in capsule formation. Like a met3 mutant, the met6 mutant was avirulent. In contrast to a met3 mutant, the met6 mutant was hypersensitive to fluconazole and to the calcineurin inhibitors FK506 and cyclosporin A. A synergistic fungicidal effect was also found between each of these drugs and met6. The phenotypic differences between the met3 and met6 mutants may be due to the accumulation in met6 mutants of homocysteine, a toxic metabolic intermediate that inhibits sterol biosynthesis. PMID- 15347760 TI - Cell-surface alterations in class IIa bacteriocin-resistant Listeria monocytogenes strains. AB - Strains of the food-borne pathogen Listeria monocytogenes, showing either intermediate or high-level resistance to class IIa bacteriocins, were investigated to determine characteristics that correlated with their sensitivity levels. Two intermediate and one highly resistant spontaneous mutant of L. monocytogenes B73, a highly resistant mutant of L. monocytogenes 412, and a highly resistant, defined (mptA) mutant of L. monocytogenes EGDe were compared with their respective wild-type strains in order to investigate the contribution of different factors to resistance. Decreased mannose-specific phosphotransferase system gene expression (mptA, EIIAB(Man) component) was implicated in all levels of resistance, confirming previous studies by the authors' group. However, a clear correlation between d-alanine content in teichoic acid (TA), in particular the alanine : phosphorus ratio, and a more positive cell surface, as determined by cytochrome c binding, were found for the highly resistant strains. Furthermore, two of the three highly resistant strains showed a significant increase in sensitivity towards d-cycloserine (DCS). However, real-time PCR of the dltA (d-alanine esterification), and dal and ddlA genes (peptidoglycan biosynthesis) showed no change in transcriptional levels. The link between DCS sensitivity and increased d-alanine esterification of TA may be that DCS competes with alanine for transport via the alanine transporter. A possible tendency towards increased lysinylation of membrane phospholipid in the highly resistant strains was also found. A previous study reported that cell membranes of all the resistant strains, including the intermediate resistant strains, contained more unsaturated phosphatidylglycerol, which is an indication of a more fluid cell membrane. The results of that study correlate with the possible lysinylation, decreased mptA expression, d-alanine esterification of TA and more positive cell surface charge found in this study for resistant strains. The authors' findings strongly indicate that all these factors could contribute to class IIa bacteriocin resistance and that the combination and contribution of each of these factors determine the level of bacteriocin resistance. PMID- 15347761 TI - A spiroplasma associated with tremor disease in the Chinese mitten crab (Eriocheir sinensis). AB - An epidemic of tremor disease has been a serious problem in Chinese mitten crabs, Eriocheir sinensis, in China in recent years. The disease-causing agent was previously considered to be a rickettsia-like organism. Here, analysis of the 16S rRNA gene sequence, light and electron microscopy and cultivation in vitro were used to identify the agent. Sequence analysis of the 16S rRNA gene found it to have 98 % identity with that of Spiroplasma mirum. The agent was able to be passed through membrane filters with pores 220 nm in diameter and could be cultivated by inoculating the yolk sac of embryonated chicken eggs and M1D medium. Rotary motion and flexional movement were seen by light microscopy, and electron microscopy showed that the organism had a helical morphology and lacked a cell wall. The organism produced small colonies with a diameter of 40-50 microm after 17-25 days of incubation on solid M1D medium. The agent was found in blood cells, muscles, nerves and connective tissues of crabs inoculated with a filtrate of yolk sacs or with cultures grown in M1D medium, and it was similar in structure to those grown in eggs and cultivation broth. Disease was reproduced by experimental infection with the cultivated organisms. This study has demonstrated that the causative agent of tremor disease in the Chinese mitten crab is a member of the genus Spiroplasma. This is believed to be the first time a spiroplasma has been found in a crustacean. These findings are not only significant for studies on pathogenic spiroplasmas, but also have implications for studies of freshwater ecology. PMID- 15347762 TI - Identification of the dialysable serum inducer of germ-tube formation in Candida albicans. AB - Yeast cells of Candida albicans are induced by serum at 37 degrees C to produce germ tubes, the first step in a transition from yeast to hyphal growth. Previously, it has been shown that the active component is not serum albumin but is present in the dialysable fraction of serum. In this study, serum induction of germ-tube formation is shown to occur even in the presence of added exogenous nitrogen sources and is therefore not signalled by nitrogen derepression. The active component in serum was purified by ion-exchange, reverse-phase and size exclusion chromatography from the dialysable fraction of serum and was identified by NMR to be d-glucose. Enzymic destruction of glucose, using glucose oxidase, demonstrated that d-glucose was the only active component in these fractions. Induction of germ-tube formation by d-glucose required a temperature of 37 degrees C and the pH optimum was between pH 7.0 and 8.0. d-Glucose induced germ tube formation in a panel of clinical isolates of C. albicans. Although d-glucose is the major inducer in serum, a second non-dialysable, trichloroacetic acid precipitable inducer is also present. However, whereas either 1.4 % (v/v) serum or an equivalent concentration of d-glucose induced 50 % germ-tube formation, the non-dialysable component required a 10-fold higher concentration to induce 50 % germ-tube formation. Serum is, therefore, the most effective induction medium for germ-tube formation because it is buffered at about pH 8.5 and contains two distinct inducers (glucose and a non-dialysable component), both active at this pH. PMID- 15347763 TI - Complex regulation of the synthesis of the compatible solute ectoine in the halophilic bacterium Chromohalobacter salexigens DSM 3043T. AB - The synthesis of the compatible solute ectoine, mediated by the ectABC gene products, is the main mechanism used by the halophilic bacterium Chromohalobacter salexigens to cope with osmotic stress. Evidence was found that this process is regulated at the transcriptional level. S1 protection analyses performed with RNA extracted from cells grown in minimal medium at low (0.75 M NaCl) or high (2.5 M NaCl) osmolarity suggested the existence of four promoters upstream of ectA. Two of these (PectA1 and PectA2) might be recognized by the main vegetative sigma factor sigma(70), and one (PectA3) might be dependent on the general stress sigma factor sigma(S). The S1 protection assays suggest that PectA1 and PectA3 may be osmoregulated promoters. In addition, an internal promoter showing sequences homologous to promoters dependent on the heat-shock sigma factor sigma(32) was found upstream of ectB. Transcription from PectA in C. salexigens followed a pattern typical of sigma(S)-dependent promoters, and was reduced by 50 % in an E. coli rpoS background. These data strongly suggest the involvement of the general stress sigma factor sigma(S) in ectABC transcription in C. salexigens. Expression of PectA-lacZ and PectB-lacZ trancriptional fusions was very high at low salinity, suggesting that ectABC may be a partially constitutive system. Both transcriptional fusions were induced during continuous growth at high temperature and their expression was reduced in cells grown in the presence of osmoprotectants (ectoine or glycine betaine) or the DNA gyrase inhibitor nalidixic acid. Moreover, PectA-lacZ expression was negatively modulated in cells grown with an excess of iron (FeCl(3)). Measurement of ectoine levels in the presence of glycine betaine at different NaCl concentrations suggests that an additional post-transcriptional control may occur as well. PMID- 15347764 TI - Expression of Melanocarpus albomyces laccase in Trichoderma reesei and characterization of the purified enzyme. AB - Previous studies on Melanocarpus albomyces laccase have shown that this enzyme is very interesting for both basic research purposes and industrial applications. In order to obtain a reliable and efficient source for this laccase, it was produced in the filamentous fungus Trichoderma reesei. Two approaches were used: production of a non-fused laccase and a hydrophobin-laccase fusion protein. Both proteins were expressed in T. reesei under the cbh1 promoter, and significantly higher activities were obtained with the non-fused laccase in shake-flask cultures (corresponding to about 230 mg l(-1)). Northern blot analyses showed rather similar mRNA levels from both expression constructs. Western analysis indicated intracellular accumulation and degradation of the hydrophobin-laccase fusion protein, showing that production of the fusion was limited at the post transcriptional level. No induction of the unfolded protein response pathway by laccase production was detected in the transformants by Northern hybridization. The most promising transformant was grown in a fermenter in batch and fed-batch modes. The highest production level obtained in the fed-batch culture was 920 mg l(-1). The recombinant laccase was purified from the culture supernatant after cleaving the major contaminating protein, cellobiohydrolase I, by papain. The recombinant and wild-type laccases were compared with regard to substrate kinetics, molecular mass, pH optimum, thermostability, and processing of the N- and C-termini, and they showed very similar properties. PMID- 15347765 TI - A linear megaplasmid, p1CP, carrying the genes for chlorocatechol catabolism of Rhodococcus opacus 1CP. AB - The Gram-positive actinobacterium Rhodococcus opacus 1CP is able to utilize several (chloro)aromatic compounds as sole carbon sources, and gene clusters for various catabolic enzymes and pathways have previously been identified. Pulsed field gel electrophoresis indicates the occurrence of a 740 kb megaplasmid, designated p1CP. Linear topology and the presence of covalently bound proteins were shown by the unchanged electrophoretic mobility after S1 nuclease treatment and by the immobility of the native plasmid during non-denaturing agarose gel electrophoresis, respectively. Sequence comparisons of both termini revealed a perfect 13 bp terminal inverted repeat (TIR) as part of an imperfect 583/587 bp TIR, as well as two copies of the highly conserved centre (GCTXCGC) of a palindromic motif. An initial restriction analysis of p1CP was performed. By means of PCR and hybridization techniques, p1CP was screened for several genes encoding enzymes of (chloro)aromatic degradation. A single maleylacetate reductase gene macA, the clc gene cluster for 4-chloro-/3,5-dichlorocatechol degradation, and the clc2 gene cluster for 3-chlorocatechol degradation were found on p1CP whereas the cat and pca gene clusters for the catechol and the protocatechuate pathways, respectively, were not. Prolonged cultivation of the wild-type strain 1CP under non-selective conditions led to the isolation of the clc- and clc2-deficient mutants 1CP.01 and 1CP.02 harbouring the shortened plasmid variants p1CP.01 (500 kb) and p1CP.02 (400 kb). PMID- 15347766 TI - Vasopressin mechanism-mediated pressor responses caused by central angiotensin II in the ovine fetus. AB - AVP not only influences renal water excretion but also has profound cardiovascular effects in adults. Our recent studies have demonstrated that central angiotensin induced fetal pressor responses accompanied with AVP release. However, little is known of hormonal mechanisms in angiotensin-mediated fetal blood pressure (BP) changes. The present study determined AVP mechanisms in central angiotensin-mediated fetal pressor responses. The V1-receptor antagonist or V2-receptor antagonist was infused intravenously into the ovine fetus at 90% gestation. Angiotensin II (Ang II; 1.5 microg/kg) was then injected intracerebroventricularly into the chronically instrumented fetus. Ang II produced a significant increase in fetal systolic, diastolic, and mean arterial pressure adjusted to amniotic pressure (A-MAP). The enhanced fetal A-MAP was associated with intense c-fos expression in the central putative cardiovascular area: the paraventricular nuclei (PVN). Double labeling demonstrated that a number of the AVP-containing neurons in the PVN were expressing c-fos in response to central Ang II. Consistent with the activation of AVP neurons in the PVN, fetal plasma AVP was markedly enhanced. Fetal i.v. V1-receptor antagonist or V2 receptor antagonist had no effect on either fetal or maternal baseline BP. However, intracerebroventricular Ang II-increased BP was partially inhibited, although not completely abolished, by the V1-receptor blockade. In contrast, fetal i.v. infusion of V2-receptor antagonist had no effect on the pressor responses induced by central Ang II. The results suggest that the central Ang II mediated pressor responses at the last third of gestation is mediated partially by the AVP mechanism via V1 not V2 receptors. PMID- 15347767 TI - Increased poliovirus-specific intestinal antibody response coincides with promotion of Bifidobacterium longum-infantis and Bifidobacterium breve in infants: a randomized, double-blind, placebo-controlled trial. AB - To determine whether the size of the intestinal bifidobacterial population can influence the immune response to poliovirus vaccination in infants, we set up a randomized, placebo-controlled trial. From birth to 4 mo, infants were given a fermented infant formula (FIF) or a standard formula (placebo). Bifidobacteria were quantified monthly in infant stools. Antipoliovirus IgA response to Pentacoq was assessed before and 1 mo after the second vaccine injection. Thirty infants were randomized, and 20 completed the study (nine in the placebo group and 11 in the FIF group). Fecal bifidobacterial level was significantly higher with the FIF group at 4 mo of age (p=0.0498). Furthermore, B. longum/B. infantis carriage was higher at 4 mo in the FIF group (p=0.0399). Antipoliovirus IgA titers increased after Pentacoq challenge (p <0.001), and the rise was significantly higher in the FIF group (p <0.02). Antibody titers correlated with bifidobacteria, especially with B. longum/B. infantis and B. breve levels (p <0.002). Infants who harbored B. longum/B. infantis also exhibited higher levels of antipoliovirus IgAs (p <0.002). In conclusion, the present results indicate that antipoliovirus response can be triggered with a fermented formula that is able to favor intestinal bifidobacteria. Whether this effect on the immune system is achieved through the bifidogenic effect of the formula (mainly through B. longum/B. infantis and B. breve stimulation) or directly linked to compounds (i.e. peptides) produced by milk fermentation remains to be investigated. PMID- 15347768 TI - Mitochondrial fatty acid beta-oxidation in the human eye and brain: implications for the retinopathy of long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency. AB - The retinal pigment epithelium (RPE) and the choriocapillaris are affected early in the retinopathy associated with long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency. RPE in culture possesses the machinery needed for mitochondrial fatty acid beta-oxidation in vitro. To further elucidate pathogenesis of LCHAD retinopathy, we performed immunohistochemistry of the human eye and brain with antibodies to beta-oxidation enzymes. Human eye and brain sections were stained with antibodies to medium-chain (MCAD) and very long-chain acyl-CoA dehydrogenase (VLCAD), short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD), and mitochondrial trifunctional protein (MTP) harboring LCHAD. Antibodies to 2-methyl-3-hydroxybutyryl-CoA dehydrogenase (MHBD) and cytochrome c oxidase subunit I (COX I) were used as a reference. VLCAD, MTP, MCAD, SCHAD, MHBD, and COX I antibodies labeled most retinal layers and tissues of the human eye actively involved in oxidative metabolism (extraocular and intraocular muscle, the RPE, the corneal endothelium, and the ciliary epithelium). MTP and COX I antibodies labeled the inner segments of photoreceptors. The choriocapillaris was labeled only with SCHAD and MCAD antibodies. In the brain, the choroid plexus and nuclei of the brain stem were most intensely labeled with beta-oxidation antibodies, whereas COX I antibodies strongly labeled neurons in several regions of the brain. Mitochondrial fatty acid beta-oxidation likely plays a role in ocular energy production in vivo. The RPE rather than the choriocapillaris could be the critical affected cell layer in LCHAD retinopathy. Reduced energy generation in the choroid plexus may contribute to the cerebral edema observed in patients with beta-oxidation defects. PMID- 15347769 TI - Blood transfusion increases functional capillary density in the skin of anemic preterm infants. AB - Direct visualization of the microcirculation at the level of the skin capillaries may provide information on the quality of tissue perfusion. Orthogonal polarization spectral imaging (OPS) enables noninvasively direct observation of those blood vessels. OPS was applied to the upper arm of 13 preterm anemic infants [median (95% confidence interval) gestational age: 26 wk (25-26 wk); birth weight: 730 g (652-789 g)] before and 2 and 24 h after transfusion (Tx). OPS images of skin perfusion were continuously recorded on video. Off-line quantitative data of microvascular perfusion were obtained by measuring functional capillary density, vessel diameter, red blood cell velocity, and flow. We found a significant increase in functional capillary density 2 h after transfusion with an additional significant rise after 24 h [before: 142 (134 155); 2 h after Tx: 185 (166-196); 24 h after Tx: 206 (185-219) cm/cm2; p < 0.001), thus indicating improved microvascular perfusion. Vessel diameter, red blood cell velocity, and flow did not change significantly. There were no significant changes in clinical variables, such as blood pressure, heart rate, or body temperature. Whereas conventional monitoring methods did not show any changes after transfusion, quantitative analyses of OPS images indicated improved perfusion; hence, it seems a useful monitor for assessing the response to therapies aimed to improve tissue perfusion. PMID- 15347770 TI - Complement activation reflects severity of meconium aspiration syndrome in newborn pigs. AB - Meconium aspiration syndrome (MAS) is a serious condition in newborns, associated with a poorly characterized inflammatory reaction. The aim of this study was to investigate a possible role for complement in pulmonary pathophysiology and systemic inflammation in experimental MAS. MAS was induced by instillation of meconium into the lungs of 12 hypoxic piglets. Six controls received saline under otherwise identical conditions. Hemo- and lung dynamics were recorded for 5 h. Plasma complement activation, revealed by the terminal sC5b-9 complex (TCC), and cytokines were measured by enzyme immunoassays. TCC increased substantially in MAS animals compared with controls (p <0.0005). The increase in TCC correlated with lung dysfunction: closely with oxygenation index (r=0.51, p <0.0001) and ventilation index (r=0.64, p < 0.0001) and inversely with lung compliance (r= 0.22, p=0.05). IL-1beta and tumor necrosis factor-alpha increased significantly in MAS animals compared with the controls (p=0.004 and 0.008, respectively). The cytokine increase occurred later than TCC and showed correlations with lung dysfunction similar to TCC. IL-10 did not discriminate between MAS animals and controls (p=0.32). Finally, the subgroup of MAS animals that died (n=5) had substantially higher TCC concentration compared with the surviving MAS animals (n=7; p <0.0005). TCC increased substantially in MAS and was closely correlated to lung dysfunction. Complement activation preceded cytokine release, which may suggest a primary role for complement in the pathophysiology of MAS. PMID- 15347771 TI - Disease-specific markers for the mucopolysaccharidoses. AB - Unprecedented demands are now placed on clinicians for early diagnosis as we enter into an era of advancing treatment opportunities for the mucopolysaccharidoses (MPS). Biochemical monitoring of any therapeutic avenue will also be prerequisite. To this end, we aimed to identify a range of urinary oligosaccharides that could be used to identify and characterize patients with MPS. We analyzed 94 urine samples from 68 patients with MPS and 26 control individuals for oligosaccharides derived from glycosaminoglycans using electrospray ionization-tandem mass spectrometry. The oligosaccharide profile for each patient group was compared with that of the control group. The Mann-Whitney U test was used to measure the difference between each patient group and the controls for each analyte. Urine samples from patients before and at successive times after bone marrow transplantation were also evaluated. A number of oligosaccharides were identified in the urine of each MPS subtype, and for each of these, specific oligosaccharide profiles were formulated. These profiles enabled the identification of all 68 patients and their subtypes with the exception of MPS IIIB and IIIC. Selected oligosaccharides were used to assess three individuals after a bone marrow transplant, and, in each case, a substantial reduction in the level of diagnostic oligosaccharides, posttransplantation, was observed. The identification and measurement of glycosaminoglycan-derived oligosaccharides in urine provides a sensitive and specific screen for the early identification of individuals with MPS. The resulting oligosaccharide profiles not only characterize subtype but also provide a disease-specific fingerprint for the biochemical monitoring of current and proposed therapies. PMID- 15347772 TI - Resuscitation with 100% O2 increases cerebral injury in hypoxemic piglets. AB - Perinatal asphyxia is a major cause of immediate and postponed brain injury in the newborn. We hypothesized that resuscitation with 100% O2 compared with ambient air is detrimental to the cerebral tissue. We assessed cerebral injury in newborn piglets that underwent global hypoxia and subsequent resuscitation with 21 or 100% O2 by extracellular glycerol, matrix metalloproteinase (MMP) expression levels, and oxidative stress. Extracellular glycerol was sampled by cerebral microdialysis. MMP levels were analyzed in cerebral tissue by gelatin zymography, broad matrix degrading capacity, and real-time PCR. Total endogenous antioxidant capacity was measured by the oxygen radical absorbance capacity assay. Extracellular glycerol increased 50% after resuscitation with 100% O2 compared with 21% O2. Total MMP activity was doubled in resuscitated animals at endpoint compared with baseline (p=0.018), and the MMP-2 activity was significantly increased in piglets that were resuscitated with 21% O(2) (p=0.003) and 100% O2 (p=0.001) compared with baseline. MMP-2 mRNA level was 100% increased in piglets that were resuscitated with 100% O2 as compared with 21% O2 (p < 0.05). Oxygen radical absorbance capacity values in piglets that were resuscitated with 100% O2 were considerably reduced compared with both baseline (p=0.001) and piglets that were resuscitated with 21% O2 (p=0.001). In conclusion, our data show increased MMP-2 activity at both gene and protein levels, accompanied with cerebral leakage of glycerol, presumably triggered by augmented oxidative stress. Our findings suggest that resuscitation of asphyxiated piglets with 100% O2 is detrimental to the piglet brain compared with resuscitation with 21% O2. PMID- 15347773 TI - Effects of vitamin E supplementation on intracellular antioxidant enzyme production in adolescents with type 1 diabetes and early microangiopathy. AB - Defective intracellular antioxidant enzyme production (IAP) has been demonstrated in adults with diabetic nephropathy. To evaluate the effects on IAP of vitamin E administration in adolescents with type 1 diabetes and early signs of microangiopathy, 12 adolescents (aged 11-21 y; diabetes duration 10-18) were studied. Eight had retinopathy [background (four), preproliferative (three), or proliferative (one)], four had persistent microalbuminuria, and seven had both. Skin fibroblasts were obtained by biopsies and cultured in Dulbecco's modified Eagle's medium. CuZn superoxide dismutase (SOD), MnSOD, catalase (CAT), and glutathione-peroxidase (GPX) activity and mRNA expression were measured before and after 3 mo of synthetic vitamin E supplementation (600 mg twice daily); on both occasions, IAP was evaluated at different ex vivo glucose concentrations (5 and 22 mM). Ten adolescents with type 1 diabetes (aged 12-20 y) without angiopathy and eight healthy volunteers (aged 15-22 y) participated as control subjects. Vitamin E serum levels were measured throughout the study. In normal glucose concentrations, CuZnSOD, MnSOD, CAT, and GPX activity and mRNA expression were not different among the groups. In high glucose, CuZnSOD activity and mRNA increased similarly in all groups [angiopathics: 0.96 +/- 0.30 U/mg protein; 9.9 +/- 3.2 mRNA/glyceraldehyde-3-phosphate dehydrogenase). CAT and GPX activity and mRNA did not increase in high glucose only in adolescents with angiopathy (0.35 +/- 0.09; 4.2 +/- 0.1 and 0.52 +/- 0.14; 2.4 +/- 0.9, respectively). MnSOD did not change in any group. Vitamin E supplementation had no effect on any enzymatic activity and mRNA in both normal and hyperglycemic conditions. Adolescents with early signs of diabetic angiopathy have defective IAP and activity, which are not modified by vitamin E. PMID- 15347774 TI - How sedentary are people in "sedentary" occupations? The physical activity of teachers in urban South India. AB - AIMS: The aim of this study was to assess the total and occupational physical activity status of school and college teachers in Bangalore, South India and to evaluate the use of "job description" as an indicator of occupational physical activity. METHOD: A self-administered questionnaire was given to a convenience sample of 198 (25 males and 173 females) school and college teachers. RESULTS: Despite the categorization of "teacher" in India as "sedentary", our data indicate that the overall physical activity status of teachers varies considerably and is dependent on non-occupational physical activity, such as household chores and discretionary exercise. Thus, only 12% of teachers were truly sedentary, based on computations of the contributions of all measured physical activity domains, while 10% were heavily active. The contribution of other physical activity domains to the overall physical activity status in our data set was dependent on gender; in women, the energy expenditure related to household chores was 4-fold higher than in men (P < 0.01), while in men, energy expenditure related to discretionary exercise was 3-fold higher than in women (P < 0.01). CONCLUSIONS: This study highlights the problems associated with extrapolating occupational activity using job descriptors to overall physical activity status. The findings have implications in epidemiological studies which use occupational activity as a surrogate for overall physical activity status. However, the impact of potential misclassifications will need to be assessed within a socio-cultural context. PMID- 15347775 TI - Silicosis and lung cancer: a mortality study of compensated men with silicosis in New South Wales, Australia. AB - BACKGROUND: Lung cancer mortality has been found to be in excess in several groups with silicosis, but allowance for smoking was not always possible. We investigated the lung cancer mortality in men with silicosis in New South Wales, Australia, who were compensated, making allowance for smoking habits. METHODS: A mortality study of 1467 men with silicosis in New South Wales who were compensated was carried out comparing observed mortality with that expected from the New South Wales death rates adjusting for age and period. Their smoking habits were compared with national survey smoking rates and the expected number of lung cancer deaths adjusted for smoking. Cases were coded for occupation and industry. RESULTS: The observed mortality was higher than expected, but the only site of cancer showing a significant excess was the lung. The group with silicosis had smoked more than the national rates. After adjusting for smoking the standardized mortality ratio for lung cancer was 1.9 (95% confidence interval 1.5-2.3). Although there were differences in lung cancer mortality between industries and occupations, these differences were not statistically significant. CONCLUSIONS: The excess lung cancer death rate may not be entirely due to silica exposure because compensation may have been influenced by the presence of chronic obstructive respiratory disease and there is some evidence that the presence of this disease increases lung cancer risk independently of smoking. PMID- 15347776 TI - Ill-health retirement: a survey of decision making by occupational physicians working for local authority fire and police services. AB - AIMS: To investigate the effect of qualifications, experience and service on ill health retirement decisions made by police force and fire brigade medical advisers. METHODS: Doctors advising UK fire and police authorities attending a conference were given 16 case summaries from experienced fire and police advisers and asked to predict the outcome in terms of return to work or ill-health retirement. The answers were scored by assessing whether the outcome predicted by the participant agreed with the case setter. Correlation coefficients were calculated and a kappa coefficient was determined for assessing inter-participant variation. RESULTS: Forty-seven doctors attended, 44 participated and the responses of 39 (86%) were analysed. Over half the doctors participating had held their post for less than 5 years. The majority (>50%) agreed with the case setter in 11 case summaries and in five cases the agreement was strong (>75%). The majority disagreed in five cases, with < or =80% dissenting in three cases. There was wide variation in the inter-participant variation (kappa = -0.333 to +0.5) but the total agreed score correlated significantly (P < 0.05) with experience and service as a fire or police adviser (kappa = 0.33) and ill-health retirement agreement correlated significantly with experience (kappa = 0.49). There was no correlation between the level of qualification and total correct decisions. CONCLUSIONS: Whilst we found general agreement overall, our results suggest that there is inconsistency in ill-health retirement decision making by police and fire service medical advisers in certain cases. Experience appears to improve the likelihood of agreement in decision making. PMID- 15347777 TI - Surveillance of Australian workplace Based Respiratory Events (SABRE): notifications for the first 3.5 years and validation of occupational asthma cases. AB - AIMS: In Australia, the SABRE programme, a notification scheme, has been established to collect incidence data on occupational lung disease. This paper reports the first 3.5 years of this scheme and the results of an occupational asthma validation study. METHODS: A notification form is mailed regularly to thoracic physicians and occupational physicians in the Australian states of Victoria and Tasmania, who use this to report new cases of occupational lung disease. The validation study was performed by a blinded panel of two doctors, who reviewed information extracted from the medical files of a sample of reported cases of occupational asthma. RESULTS: A total of 520 diagnoses were reported. The mean (+/- SD) age of the 448 patients notified was 55.7 +/- 16.2 years. There were 394 (88%) males and 54 females. The most common single condition was asthma for which the most common causative agent was wood dust. However, the most commonly reported agent overall was asbestos. The validation study of occupational asthma found only fair agreement (kappa = 0.4) between the panel and notifying doctors. However, agreement was better (kappa = 0.5) when the analysis was restricted to those cases where the reporting doctor considered the likelihood of the diagnosis was high. CONCLUSIONS: Occupational asthma is the most common occupational respiratory condition reported, which suggests increasing importance for this disease over more traditional forms of occupational lung disease. The validation study suggests that such schemes should restrict notifications only to those cases where the likelihood of the diagnosis is considered high. PMID- 15347778 TI - Job satisfaction and psychological health of doctors at the University of Benin Teaching Hospital. AB - BACKGROUND: Studies on job satisfaction and health have mainly been carried out among Caucasian populations. Similar studies are lacking in Nigeria. AIM: To investigate the level of job satisfaction and its relationship to psychological disorder amongst Nigerian doctors. METHODS: All 190 doctors at the University of Benin Teaching Hospital were invited to participate in a cross-sectional study. The respondents completed two self-administered questionnaires, a specifically designed (25-item) questionnaire and a standard instrument--the General Health Questionnaire (GHQ 28). RESULTS: One hundred and fifty-two doctors responded, giving a response rate of 80%. Twenty-one (14%) doctors had a GHQ score of 4 and above, and were at increased likelihood of having a psychological disorder. Fifty four per cent were either very dissatisfied or dissatisfied with their jobs, while 30% were very satisfied or satisfied with their jobs. The proportion of doctors with GHQ scores of 4 or above increased with the level of dissatisfaction although this was not statistically significant. The crude odds ratio of psychological disorder (GHQ score 4 and above) in dissatisfied respondents compared to satisfied respondents was 2.2 (95% confidence interval = 0.6-8.4). The odds ratio remained non-significant after controlling for the presence of adolescent children, stress outside work and length of service. CONCLUSION: This study showed a high rate of job dissatisfaction among Nigerian doctors compared to their European and North American counterparts. The reason for this is not altogether obvious. Further research is needed to determine the causes of job dissatisfaction amongst Nigerian doctors. PMID- 15347779 TI - Increase in exhaled nitric oxide in shoe and leather workers at the end of the work-shift. AB - BACKGROUND: Measurement of nitric oxide (NO) in exhaled air is a non-invasive procedure for assessing airway or pulmonary inflammation, and for evaluating environmental irritant effects of air pollution. The objective of this study was to evaluate exhaled NO levels in synthetic leather workers exposed to organic solvents toluene, xylene and methylethyl ketone. METHODS: Seventeen shoe and leather workers and 10 controls from administrative personnel of the factory were studied. The levels of exhaled NO at baseline and at the end of the work shift were measured using a standardized chemiluminescence technique. Environmental assay of the three solvents was also assessed during the working day. RESULTS: Environmental solvent concentrations were high but within permissible exposure levels. Exhaled NO concentrations increased by 40% in the leather workers at the end of the working day from a mean +/- SD of 9.1 +/- 1.3 p.p.b. to 12.8 +/- 1.7 p.p.b. (P < 0.02), as compared to the controls. CONCLUSION: Exhaled NO is increased in shoe and leather workers exposed to solvents at the end of work shift, while it does not change in unexposed subjects of the same factory. Exhaled NO may be a sensitive tool to monitor sub-clinical effects of occupational pro-inflammatory substances. PMID- 15347780 TI - Case-control study of multiple chemical sensitivity, comparing haematology, biochemistry, vitamins and serum volatile organic compound measures. AB - BACKGROUND: Multiple chemical sensitivity (MCS), although poorly understood, is associated with considerable morbidity. AIM: To investigate potential biological mechanisms underlying MCS in a case-control study. METHODS: Two hundred and twenty-three MCS cases and 194 controls (urban females, aged 30-64 years) fulfilled reproducible eligibility criteria with discriminant validity. Routine laboratory results and serum levels of volatile organic compounds (VOCs) were compared. Dose-response relationships, a criterion for causality, were examined linking exposures to likelihood of case status. RESULTS: Routine laboratory investigations revealed clinically unimportant case-control differences in means. Confounder-adjusted odds ratios (OR) showed MCS was negatively associated with lymphocyte count and total plasma homocysteine, positively associated with mean cell haemoglobin concentration, alanine aminotransferase and serum vitamin B6, and not associated with thyroid stimulating hormone, folate or serum vitamin B12. More cases than controls had detectable serum chloroform (P = 0.001) with the OR for detectability 2.78 (95% confidence interval = 1.73-4.48, P < 0.001). Chloroform levels were higher in cases. However, cases had significantly lower means of detectable serum levels of ethylbenzene, m&p-xylene, 3-methylpentane and hexane, and means of all serum levels of 1,3,5- and 1,2,3-trimethylbenzene, 2- and 3-methylpentane, and m&p-xylene. CONCLUSIONS: Our findings are inconsistent with proposals that MCS is associated with vitamin deficiency or thyroid dysfunction, but the association of lower lymphocyte counts with an increased likelihood of MCS is consistent with theories of immune dysfunction in MCS. Whether avoidance of exposures or different metabolic pathways in cases explain the observed lower VOC levels or the higher chloroform levels should be investigated. PMID- 15347781 TI - Evaluation of a treadmill test for predicting the aerobic capacity of firefighters. AB - BACKGROUND: As part of a comprehensive occupational medical program for fire departments in the USA, the National Fire Protection Association and The Fire Service Joint Labor Management Wellness/Fitness Initiative endorse a standardized submaximal test that uses the Gerkin treadmill protocol for predicting the maximal oxygen uptake (VO(2max)) of firefighters. AIMS: To test the validity of the Gerkin treadmill protocol in healthy men and women. METHODS: Fifty-four healthy men and women (age range 19-58 years) performed the Gerkin test and a treadmill run test to maximal exhaustion. Their heart rates were monitored continuously with an electrocardiogram during each test. During the VO(2max) test, the subjects' VO(2) was measured continuously using indirect calorimetry. RESULTS: Although the predicted and observed VO(2max) values correlated (r = 0.70, P < 0.001 and standard error of estimate = 5.98 ml/kg/min), the mean values differed (49.8 +/- 8.3 and 41.8 +/- 5.8 ml/kg/min, respectively) (P < 0.001). The VO(2max) value was overestimated in 50 (93%) participants. The overestimation was >25% in 18 (33%) participants. Gender, age and VO(2max) did not affect the Gerkin protocol's predictability of VO(2max). CONCLUSIONS: The Gerkin treadmill protocol overpredicts VO(2max) in healthy men and women and, therefore, should not be used for predicting VO(2max) in individual firefighters, particularly if VO(2max) is a criterion for inclusion or exclusion from duty. At this time, a valid treadmill running test is needed for predicting the VO(2max) value of individual firefighters. PMID- 15347782 TI - Protection of plasma membrane K+ transport by the salt overly sensitive1 Na+-H+ antiporter during salinity stress. AB - Physicochemical similarities between K(+) and Na(+) result in interactions between their homeostatic mechanisms. The physiological interactions between these two ions was investigated by examining aspects of K(+) nutrition in the Arabidopsis salt overly sensitive (sos) mutants, and salt sensitivity in the K(+) transport mutants akt1 (Arabidopsis K(+) transporter) and skor (shaker-like K(+) outward-rectifying channel). The K(+)-uptake ability (membrane permeability) of the sos mutant root cells measured electrophysiologically was normal in control conditions. Also, growth rates of these mutants in Na(+)-free media displayed wild-type K(+) dependence. However, mild salt stress (50 mm NaCl) strongly inhibited root-cell K(+) permeability and growth rate in K(+)-limiting conditions of sos1 but not wild-type plants. Increasing K(+) availability partially rescued the sos1 growth phenotype. Therefore, it appears that in the presence of Na(+), the SOS1 Na(+)-H(+) antiporter is necessary for protecting the K(+) permeability on which growth depends. The hypothesis that the elevated cytoplasmic Na(+) levels predicted to result from loss of SOS1 function impaired the K(+) permeability was tested by introducing 10 mm NaCl into the cytoplasm of a patch clamped wild-type root cell. Complete loss of AKT1 K(+) channel activity ensued. AKT1 is apparently a target of salt stress in sos1 plants, resulting in poor growth due to impaired K(+) uptake. Complementary studies showed that akt1 seedlings were salt sensitive during early seedling development, but skor seedlings were normal. Thus, the effect of Na(+) on K(+) transport is probably more important at the uptake stage than at the xylem loading stage. PMID- 15347783 TI - The capacity of green oilseeds to utilize photosynthesis to drive biosynthetic processes. AB - Seeds of many plant species are green during embryogenesis. To directly assess the influence of light on the physiological status of green oilseeds in planta, Brassica napus and soybean (Glycine max) seeds were rapidly dissected from plants growing in the light or dark. The activation state of malate dehydrogenase, which reflects reduced thioredoxin and NADP/NADPH ratios, was found to be as high in seeds exposed to light as in leaves and to decrease in the dark. Rubisco was highly activated (carbamylated) in both light and dark, most likely reflecting high seed CO(2) concentrations. Activities of Rubisco and phosphoribulokinase were sufficient to account for significant refixation of CO(2) produced during B. napus oil biosynthesis. To determine the influence of light on oil synthesis in planta, siliques on intact plants in full sunlight or detached siliques fed (3)H(2)O were partly covered with aluminum foil. Seeds from light and dark sections were analyzed, and fatty acid accumulation was found to be higher in seeds exposed to light than seeds from dark sections. The spectrum of light filtering through silique walls and the pigment composition of developing B. napus embryos were determined. In addition to a low chlorophyll a/b ratio, the carotenoid pigments of seeds can provide additional capture of the green light that filters through siliques. Together, these results demonstrate that even the low level of light reaching seeds plays a substantial role in activating light regulated enzymes, increasing fatty acid synthesis, and potentially powering refixation of CO(2). PMID- 15347784 TI - The potassium-dependent transcriptome of Arabidopsis reveals a prominent role of jasmonic acid in nutrient signaling. AB - Full genome microarrays were used to assess transcriptional responses of Arabidopsis seedlings to changing external supply of the essential macronutrient potassium (K(+)). Rank product statistics and iterative group analysis were employed to identify differentially regulated genes and statistically significant coregulated sets of functionally related genes. The most prominent response was found for genes linked to the phytohormone jasmonic acid (JA). Transcript levels for the JA biosynthetic enzymes lipoxygenase, allene oxide synthase, and allene oxide cyclase were strongly increased during K(+) starvation and quickly decreased after K(+) resupply. A large number of well-known JA responsive genes showed the same expression profile, including genes involved in storage of amino acids (VSP), glucosinolate production (CYP79), polyamine biosynthesis (ADC2), and defense (PDF1.2). Our findings highlight a novel role of JA in nutrient signaling and stress management through a variety of physiological processes such as nutrient storage, recycling, and reallocation. Other highly significant K(+) responsive genes discovered in our study encoded cell wall proteins (e.g. extensins and arabinogalactans) and ion transporters (e.g. the high-affinity K(+) transporter HAK5 and the nitrate transporter NRT2.1) as well as proteins with a putative role in Ca(2+) signaling (e.g. calmodulins). On the basis of our results, we propose candidate genes involved in K(+) perception and signaling as well as a network of molecular processes underlying plant adaptation to K(+) deficiency. PMID- 15347785 TI - The Elm1 (ZmHy2) gene of maize encodes a phytochromobilin synthase. AB - The light insensitive maize (Zea mays) mutant elongated mesocotyl1 (elm1) has previously been shown to be deficient in the synthesis of the phytochrome chromophore 3E-phytochromobilin (PPhiB). To identify the Elm1 gene, a maize homolog of the Arabidopsis PPhiB synthase gene AtHY2 was isolated and designated ZmHy2. ZmHy2 encodes a 297-amino acid protein of 34 kD that is 50% identical to AtHY2. ZmHY2 was predicted to be plastid localized and was targeted to chloroplasts following transient expression in tobacco (Nicotiana plumbaginifolia) leaves. Molecular mapping indicated that ZmHy2 is a single copy gene in maize that is genetically linked to the Elm1 locus. Sequence analysis revealed that the ZmHy2 gene of elm1 mutants contains a single G to A transition at the 3' splice junction of intron III resulting in missplicing and premature translational termination. However, flexibility in the splicing machinery allowed a small pool of in-frame ZmHy2 transcripts to accumulate in elm1 plants. In addition, multiple ZmHy2 transcript forms accumulated in both wild-type and elm1 mutant plants. ZmHy2 splice variants were expressed in Escherichia coli and products examined for activity using a coupled apophytochrome assembly assay. Only full-length ZmHY2 (as defined by homology to AtHY2) was found to exhibit PPhiB synthase activity. Thus, the elm1 mutant of maize is deficient in phytochrome response due to a lesion in a gene encoding phytochromobilin synthase that severely compromises the PPhiB pool. PMID- 15347786 TI - A second protein L-isoaspartyl methyltransferase gene in Arabidopsis produces two transcripts whose products are sequestered in the nucleus. AB - The spontaneous and deleterious conversion of l-asparaginyl and l-aspartyl protein residues to l-iso-Asp or d-Asp occurs as proteins age and is accelerated under stressful conditions. Arabidopsis (Arabidopsis L. Heynh.) contains two genes (At3g48330 and At5g50240) encoding protein-l-isoaspartate methyltransferase (EC 2.1.1.77; PIMT), an enzyme capable of correcting this damage. The gene located on chromosome 5 (PIMT2) produces two proteins differing by three amino acids through alternative 3' splice site selection in the first intron. Recombinant protein from both splicing variants has PIMT activity. Subcellular localization using cell fractionation followed by immunoblot detection, as well as confocal visualization of PIMT:GFP fusions, demonstrated that PIMT1 is cytosolic while a canonical nuclear localization signal, present in PIMT2psi and the shorter PIMT2omega, is functional. Multiplex reverse transcription-PCR was used to establish PIMT1 and PIMT2 transcript presence and abundance, relative to beta-TUBULIN, in various tissues and under a variety of stresses imposed on seeds and seedlings. PIMT1 transcript is constitutively present but can increase, along with PIMT2, in developing seeds presumably in response to increasing endogenous abscisic acid (ABA). Transcript from PIMT2 also increases in establishing seedlings due to exogenous ABA and applied stress presumably through an ABA dependent pathway. Furthermore, cleaved amplified polymorphic sequences from PIMT2 amplicons determined that ABA preferentially enhances the production of PIMT2omega transcript in leaves and possibly in tissues other than germinating seeds. PMID- 15347787 TI - Expression patterns of a novel AtCHX gene family highlight potential roles in osmotic adjustment and K+ homeostasis in pollen development. AB - A combined bioinformatic and experimental approach is being used to uncover the functions of a novel family of cation/H(+) exchanger (CHX) genes in plants using Arabidopsis as a model. The predicted protein (85-95 kD) of 28 AtCHX genes after revision consists of an amino-terminal domain with 10 to 12 transmembrane spans (approximately 440 residues) and a hydrophilic domain of approximately 360 residues at the carboxyl end, which is proposed to have regulatory roles. The hydrophobic, but not the hydrophilic, domain of plant CHX is remarkably similar to monovalent cation/proton antiporter-2 (CPA2) proteins, especially yeast (Saccharomyces cerevisiae) KHA1 and Synechocystis NhaS4. Reports of characterized fungal and prokaryotic CPA2 indicate that they have various transport modes, including K(+)/H(+) (KHA1), Na(+)/H(+)-K(+) (GerN) antiport, and ligand-gated ion channel (KefC). The expression pattern of AtCHX genes was determined by reverse transcription PCR, promoter-driven beta-glucuronidase expression in transgenic plants, and Affymetrix ATH1 genome arrays. Results show that 18 genes are specifically or preferentially expressed in the male gametophyte, and six genes are highly expressed in sporophytic tissues. Microarray data revealed that several AtCHX genes were developmentally regulated during microgametogenesis. An exciting idea is that CHX proteins allow osmotic adjustment and K(+) homeostasis as mature pollen desiccates and then rehydrates at germination. The multiplicity of CHX-like genes is conserved in higher plants but is not found in animals. Only 17 genes, OsCHX01 to OsCHX17, were identified in rice (Oryza sativa) subsp. japonica, suggesting diversification of CHX in Arabidopsis. These results reveal a novel CHX gene family in flowering plants with potential functions in pollen development, germination, and tube growth. PMID- 15347788 TI - LWR1 and LWR2 are required for osmoregulation and osmotic adjustment in Arabidopsis. AB - With the goal of identifying molecular components of the low-water-potential response, we have carried out a two-part selection and screening strategy to identify new Arabidopsis mutants. Using a system of polyethylene glycol-infused agar plates to impose a constant low-water-potential stress, putative mutants impaired in low-water-potential induction of the tomato (Lycopersicon esculentum) le25 promoter were selected. These lines were then screened for altered accumulation of free Pro. The seedlings of 22 mutant lines had either higher or lower Pro content than wild type when exposed to low water potential. Two mutants, designated low-water-potential response1 (lwr1) and lwr2, were characterized in detail. In addition to higher Pro accumulation, lwr1 seedlings had higher total solute content, greater osmotic adjustment at low water potential, altered abscisic acid content, and increased sensitivity to applied abscisic acid with respect to Pro content. lwr1 also had altered growth and morphology. lwr2, in contrast, had lower Pro content and less osmotic adjustment leading to greater water loss at low water potential. Both lwr1 and lwr2 also had altered leaf solute content and water relations in unstressed soil-grown plants. In both mutants, the effects on solute content were too large to be explained by the changes in Pro content alone, indicating that LWR1 and LWR2 affect multiple aspects of cellular osmoregulation. PMID- 15347789 TI - Plastid-expressed betaine aldehyde dehydrogenase gene in carrot cultured cells, roots, and leaves confers enhanced salt tolerance. AB - Salinity is one of the major factors that limits geographical distribution of plants and adversely affects crop productivity and quality. We report here high level expression of betaine aldehyde dehydrogenase (BADH) in cultured cells, roots, and leaves of carrot (Daucus carota) via plastid genetic engineering. Homoplasmic transgenic plants exhibiting high levels of salt tolerance were regenerated from bombarded cell cultures via somatic embryogenesis. Transformation efficiency of carrot somatic embryos was very high, with one transgenic event per approximately seven bombarded plates under optimal conditions. In vitro transgenic carrot cells transformed with the badh transgene were visually green in color when compared to untransformed carrot cells, and this offered a visual selection for transgenic lines. BADH enzyme activity was enhanced 8-fold in transgenic carrot cell cultures, grew 7-fold more, and accumulated 50- to 54-fold more betaine (93-101 micromol g(-1) dry weight of beta Ala betaine and Gly betaine) than untransformed cells grown in liquid medium containing 100 mm NaCl. Transgenic carrot plants expressing BADH grew in the presence of high concentrations of NaCl (up to 400 mm), the highest level of salt tolerance reported so far among genetically modified crop plants. BADH expression was 74.8% in non-green edible parts (carrots) containing chromoplasts, and 53% in proplastids of cultured cells when compared to chloroplasts (100%) in leaves. Demonstration of plastid transformation via somatic embryogenesis utilizing non green tissues as recipients of foreign DNA for the first time overcomes two of the major obstacles in extending this technology to important crop plants. PMID- 15347790 TI - Induction and functional analysis of two reduced nicotinamide adenine dinucleotide phosphate-dependent glutathione peroxidase-like proteins in Synechocystis PCC 6803 during the progression of oxidative stress. AB - Synechocystis PCC 6803 contains two types of glutathione peroxidase-like proteins (GPX-1 and GPX-2) that utilize NADPH but not reduced glutathione and unsaturated fatty acid hydroperoxides or alkyl hydroperoxides. The steady-state transcript level of gpx-1 gradually increased under oxidative stress conditions imposed by high light intensity, high salinity, or application of methylviologen or t-butyl hydroperoxide in the wild-type and GPX-2 knock-out mutant (gpx-2Delta) cells. To examine the ability of GPX-1, GPX-2, and thioredoxin peroxidase to scavenge lipid hydroperoxide in vivo, we measured the photosynthetic evolution of O(2) and the level of lipid peroxidation in the wild-type and each type of mutant cell after the application of t-butyl hydroperoxide or H(2)O(2). The data reported here indicate that GPX-1 and GPX-2 are essential for the removal of lipid hydroperoxides under normal and stress conditions, leading to the protection of membrane integrity. PMID- 15347791 TI - The fast and transient transcriptional network of gravity and mechanical stimulation in the Arabidopsis root apex. AB - Plant root growth is affected by both gravity and mechanical stimulation (Massa GD, Gilroy S [2003] Plant J 33: 435-445). A coordinated response to both stimuli requires specific and common elements. To delineate the transcriptional response mechanisms, we carried out whole-genome microarray analysis of Arabidopsis root apices after gravity stimulation (reorientation) and mechanical stimulation and monitored transcript levels of 22,744 genes in a time course during the first hour after either stimulus. Rapid, transient changes in the relative abundance of specific transcripts occurred in response to gravity or mechanical stimulation, and these transcript level changes reveal clusters of coordinated events. Transcriptional regulation occurs in the root apices within less than 2 min after either stimulus. We identified genes responding specifically to each stimulus as well as transcripts regulated in both signal transduction pathways. Several unknown genes were specifically induced only during gravitropic stimulation (gravity induced genes). We also analyzed the network of transcriptional regulation during the early stages of gravitropism and mechanical stimulation. PMID- 15347792 TI - Diurnal changes in the transcriptome encoding enzymes of starch metabolism provide evidence for both transcriptional and posttranscriptional regulation of starch metabolism in Arabidopsis leaves. AB - To gain insight into the synthesis and functions of enzymes of starch metabolism in leaves of Arabidopsis L. Heynth, Affymetrix microarrays were used to analyze the transcriptome throughout the diurnal cycle. Under the conditions employed, transitory leaf starch is degraded progressively during a 12-h dark period, and then accumulates during the following 12-h light period. Transcripts encoding enzymes of starch synthesis changed relatively little in amount over 24 h except for two starch synthases, granule bound starch synthase and starch synthase II, which increased appreciably during the transition from dark to light. The increase in RNA encoding granule-bound starch synthase may reflect the extensive destruction of starch granules in the dark. Transcripts encoding several enzymes putatively involved in starch breakdown showed a coordinated decline in the dark followed by rapid accumulation in the light. Despite marked changes in their transcript levels, the amounts of some enzymes of starch metabolism do not change appreciably through the diurnal cycle. Posttranscriptional regulation is essential in the maintenance of amounts of enzymes and the control of their activities in vivo. Even though the relationships between transcript levels, enzyme activity, and diurnal metabolism of starch metabolism are complex, the presence of some distinctive diurnal patterns of transcripts for enzymes known to be involved in starch metabolism facilitates the identification of other proteins that may participate in this process. PMID- 15347793 TI - Microbial products trigger amino acid exudation from plant roots. AB - Plants naturally cycle amino acids across root cell plasma membranes, and any net efflux is termed exudation. The dominant ecological view is that microorganisms and roots passively compete for amino acids in the soil solution, yet the innate capacity of roots to recover amino acids present in ecologically relevant concentrations is unknown. We find that, in the absence of culturable microorganisms, the influx rates of 16 amino acids (each supplied at 2.5 microm) exceed efflux rates by 5% to 545% in roots of alfalfa (Medicago sativa), Medicago truncatula, maize (Zea mays), and wheat (Triticum aestivum). Several microbial products, which are produced by common soil microorganisms such as Pseudomonas bacteria and Fusarium fungi, significantly enhanced the net efflux (i.e. exudation) of amino acids from roots of these four plant species. In alfalfa, treating roots with 200 microm phenazine, 2,4-diacetylphloroglucinol, or zearalenone increased total net efflux of 16 amino acids 200% to 2,600% in 3 h. Data from (15)N tests suggest that 2,4-diacetylphloroglucinol blocks amino acid uptake, whereas zearalenone enhances efflux. Thus, amino acid exudation under normal conditions is a phenomenon that probably reflects both active manipulation and passive uptake by microorganisms, as well as diffusion and adsorption to soil, all of which help overcome the innate capacity of plant roots to reabsorb amino acids. The importance of identifying potential enhancers of root exudation lies in understanding that such compounds may represent regulatory linkages between the larger soil food web and the internal carbon metabolism of the plant. PMID- 15347794 TI - LESION SIMULATING DISEASE 1 is required for acclimation to conditions that promote excess excitation energy. AB - The lsd1 mutant of Arabidopsis fails to limit the boundaries of hypersensitive cell death response during avirulent pathogen infection and initiates unchecked lesions in long day photoperiod giving rise to the runaway cell death (rcd) phenotype. We link here the initiation and propagation of rcd to the activity of photosystem II, stomatal conductance and ultimately to photorespiratory H(2)O(2). A cross of lsd1 with the chlorophyll a/b binding harvesting-organelle specific (designated cao) mutant, which has a reduced photosystem II antenna, led to reduced lesion formation in the lsd1/cao double mutant. This lsd1 mutant also had reduced stomatal conductance and catalase activity in short-day permissive conditions and induced H(2)O(2) accumulation followed by rcd when stomatal gas exchange was further impeded. All of these traits depended on the defense regulators EDS1 and PAD4. Furthermore, nonphotorespiratory conditions retarded propagation of lesions in lsd1. These data suggest that lsd1 failed to acclimate to light conditions that promote excess excitation energy (EEE) and that LSD1 function was required for optimal catalase activity. Through this regulation LSD1 can influence the effectiveness of photorespiration in dissipating EEE and consequently may be a key determinant of acclimatory processes. Salicylic acid, which induces stomatal closure, inhibits catalase activity and triggers the rcd phenotype in lsd1, also impaired acclimation of wild-type plants to conditions that promote EEE. We propose that the roles of LSD1 in light acclimation and in restricting pathogen-induced cell death are functionally linked. PMID- 15347795 TI - The pepper transcription factor CaPF1 confers pathogen and freezing tolerance in Arabidopsis. AB - An ERF/AP2-type transcription factor (CaPF1) was isolated by differential-display reverse transcription-PCR, following inoculation of the soybean pustule pathogen Xanthomonas axonopodis pv glycines 8ra, which induces hypersensitive response in pepper (Capsicum annuum) leaves. CaPF1 mRNA was induced under conditions of biotic and abiotic stress. Higher levels of CaPF1 transcripts were observed in disease-resistant tissue compared with susceptible tissue. CaPF1 expression was additionally induced using various treatment regimes, including ethephon, methyl jasmonate, and cold stress. To determine the role of CaPF1 in plants, transgenic Arabidopsis and tobacco (Nicotiana tabacum) plants expressing higher levels of CaPF1 were generated. Gene expression analyses of transgenic Arabidopsis and tobacco revealed that the CaPF1 level in transgenic plants affects expression of genes that contain either a GCC or a CRT/DRE box in their promoter regions. Furthermore, transgenic Arabidopsis plants expressing CaPF1 displayed tolerance against freezing temperatures and enhanced resistance to Pseudomonas syringae pv tomato DC3000. Disease tolerance was additionally observed in CaPF1 transgenic tobacco plants. The results collectively indicate that CaPF1 is an ERF/AP2 transcription factor in hot pepper plants that may play dual roles in response to biotic and abiotic stress in plants. PMID- 15347796 TI - Cellular and subcellular localization of endogenous nitric oxide in young and senescent pea plants. AB - The cellular and subcellular localization of endogenous nitric oxide (NO.) in leaves from young and senescent pea (Pisum sativum) plants was studied. Confocal laser scanning microscopy analysis of pea leaf sections with the fluorescent probe 4,5-diaminofluorescein diacetate revealed that endogenous NO. was mainly present in vascular tissues (xylem and phloem). Green fluorescence spots were also detected in the epidermal cells, palisade and spongy mesophyll cells, and guard cells. In senescent leaves, NO. generation was clearly reduced in the vascular tissues. At the subcellular level, by electron paramagnetic resonance spectroscopy with the spin trap Fe(MGD)(2) and fluorometric analysis with 4,5 diaminofluorescein diacetate, NO. was found to be an endogenous metabolite of peroxisomes. The characteristic three-line electron paramagnetic resonance spectrum of NO., with g = 2.05 and a(N) = 12.8 G, was detected in peroxisomes. By fluorometry, NO. was also found in these organelles, and the level measured of NO. was linearly dependent on the amount of peroxisomal protein. The enzymatic production of NO. from l-Arg (nitric oxide synthase [NOS]-like activity) was measured by ozone chemiluminiscence. The specific activity of peroxisomal NOS was 4.9 nmol NO. mg(-1) protein min(-1); was strictly dependent on NADPH, calmodulin, and BH(4); and required calcium. In senescent pea leaves, the NOS-like activity of peroxisomes was down-regulated by 72%. It is proposed that peroxisomal NO. could be involved in the process of senescence of pea leaves. PMID- 15347797 TI - Genetic elucidation of nitric oxide signaling in incompatible plant-pathogen interactions. AB - Recent experiments indicate that nitric oxide (NO) plays a pivotal role in disease resistance and several other physiological processes in plants. However, most of the current information about the function of NO in plants is based on pharmacological studies, and additional approaches are therefore required to ascertain the role of NO as an important signaling molecule in plants. We have expressed a bacterial nitric oxide dioxygenase (NOD) in Arabidopsis plants and/or avirulent Pseudomonas syringae pv tomato to study incompatible plant-pathogen interactions impaired in NO signaling. NOD expression in transgenic Arabidopsis resulted in decreased NO levels in planta and attenuated a pathogen-induced NO burst. Moreover, NOD expression in plant cells had very similar effects on plant defenses compared to NOD expression in avirulent Pseudomonas. The defense responses most affected by NO reduction during the incompatible interaction were decreased H(2)O(2) levels during the oxidative burst and a blockage of Phe ammonia lyase expression, the key enzyme in the general phenylpropanoid pathway. Expression of the NOD furthermore blocked UV light-induced Phe ammonia lyase and chalcone synthase gene expression, indicating a general signaling function of NO in the activation of the phenylpropanoid pathway. NO possibly functions in incompatible plant-pathogen interactions by inhibiting the plant antioxidative machinery, and thereby ensuring locally prolonged H(2)O(2) levels. Additionally, albeit to a lesser extent, we observed decreases in salicylic acid production, a diminished development of hypersensitive cell death, and a delay in pathogenesis related protein 1 expression during these NO-deficient plant-pathogen interactions. Therefore, this genetic approach confirms that NO is an important regulatory component in the signaling network of plant defense responses. PMID- 15347798 TI - AtHKT1 facilitates Na+ homeostasis and K+ nutrition in planta. AB - Genetic and physiological data establish that Arabidopsis AtHKT1 facilitates Na(+) homeostasis in planta and by this function modulates K(+) nutrient status. Mutations that disrupt AtHKT1 function suppress NaCl sensitivity of sos1-1 and sos2-2, as well as of sos3-1 seedlings grown in vitro and plants grown in controlled environmental conditions. hkt1 suppression of sos3-1 NaCl sensitivity is linked to higher Na(+) content in the shoot and lower content of the ion in the root, reducing the Na(+) imbalance between these organs that is caused by sos3-1. AtHKT1 transgene expression, driven by its innate promoter, increases NaCl but not LiCl or KCl sensitivity of wild-type (Col-0 gl1) or of sos3-1 seedlings. NaCl sensitivity induced by AtHKT1 transgene expression is linked to a lower K(+) to Na(+) ratio in the root. However, hkt1 mutations increase NaCl sensitivity of both seedlings in vitro and plants grown in controlled environmental conditions, which is correlated with a lower K(+) to Na(+) ratio in the shoot. These results establish that AtHKT1 is a focal determinant of Na(+) homeostasis in planta, as either positive or negative modulation of its function disturbs ion status that is manifested as salt sensitivity. K(+)-deficient growth of sos1-1, sos2-2, and sos3-1 seedlings is suppressed completely by hkt1-1. AtHKT1 transgene expression exacerbates K(+) deficiency of sos3-1 or wild-type seedlings. Together, these results indicate that AtHKT1 controls Na(+) homeostasis in planta and through this function regulates K(+) nutrient status. PMID- 15347799 TI - Two transcription factors are negative regulators of gibberellin response in the HvSPY-signaling pathway in barley aleurone. AB - SPINDLY (SPY) protein from barley (Hordeum vulgare L. cv Himalaya; HvSPY) negatively regulated GA responses in aleurone, and genetic analyses of Arabidopsis thaliana predict that SPY functions in a derepressible GA-signaling pathway. Many, if not all, GA-dependent responses require SPY protein, and to improve our understanding of how the SPY signaling pathway operates, a yeast two hybrid screen was used to identify both upstream and downstream components that might regulate the activity of the HvSPY protein. A number of proteins from diverse classes were identified using HvSPY as bait and barley cDNA libraries as prey. Two of the HvSPY-interacting (HSI) proteins were transcription factors belonging to the myb and NAC gene families, HSImyb and HSINAC. Interaction occurred via the tetratricopeptide repeat domain of HvSPY and specificity was shown both in vivo and in vitro. Messenger RNAs for these proteins were expressed differentially in many parts of the barley plant but at very low levels. Both HSImyb and HSINAC inhibited the GA(3) up-regulation of alpha-amylase expression in aleurone, both were activators of transcription in yeast, and the green fluorescent protein-HSI fusion proteins were localized in the nucleus. These results are consistent with the model that HSI transcription factors act downstream of HvSPY as negative regulators and that they in turn could activate other negative regulators, forming the HvSPY negative regulator-signaling pathway for GA response. PMID- 15347800 TI - Identification of a specific isoform of tomato lipoxygenase (TomloxC) involved in the generation of fatty acid-derived flavor compounds. AB - There are at least five lipoxygenases (TomloxA, TomloxB, TomloxC, TomloxD, and TomloxE) present in tomato (Lycopersicon esculentum Mill.) fruit, but their role in generation of fruit flavor volatiles has been unclear. To assess the physiological role of TomloxC in the generation of volatile C6 aldehyde and alcohol flavor compounds, we produced transgenic tomato plants with greatly reduced TomloxC using sense and antisense constructs under control of the cauliflower mosaic virus 35S promoter. The expression level of the TomloxC mRNA in some transgenic plants was selectively reduced by gene silencing or antisense inhibition to between 1% and 5% of the wild-type controls, but the expression levels of mRNAs for the four other isoforms were unaffected. The specific depletion of TomloxC in transgenic tomatoes led to a marked reduction in the levels of known flavor volatiles, including hexanal, hexenal, and hexenol, to as little as 1.5% of those of wild-type controls following maceration of ripening fruit. Addition of linoleic or linolenic acid to fruit homogenates significantly increased the levels of flavor volatiles, but the increase with the TomloxC depleted transgenic fruit extracts was much lower than with the wild-type control. Confocal imaging of tobacco (Nicotiana tabacum) leaf cells expressing a TomloxC-GFP fusion confirmed a chloroplast localization of the protein. Together, these results suggest that TomloxC is a chloroplast-targeted lipoxygenase isoform that can use both linoleic and linolenic acids as substrates to generate volatile C6 flavor compounds. The roles of the other lipoxygenase isoforms are discussed. PMID- 15347801 TI - Down-regulation of DELLA genes is not essential for germination of tomato, soybean, and Arabidopsis seeds. AB - The relationship between expression of a negative regulator of GA signal transduction (RGL2) belonging to the DELLA gene family and repression of Arabidopsis seed germination has been studied (Lee S, Cheng H, King KE, Wang W, He Y, Hussain A, Lo J, Harberd NP, Peng J [2002] Genes and Development 16: 646 658). There is one DELLA gene (LeGAI) present in tomato (Lycopersicon esculentum Mill.), which is expressed in both vegetative and reproductive tissues. During germination of wild-type tomato seed, there was no decline in the expression of LeGAI in either the embryo or the endosperm. Rather, LeGAI transcripts increased in these tissues following imbibition and remained high during and following germination. A similar increase in LeGAI transcripts occurred in the endosperm and embryo of GA-treated gib-1 mutant seed during and following germination. Likewise in soybean (Glycine max) seed, there was no decline in the expression of two DELLA genes in the radicle before or after germination. Upon reexamination of RGL2 in Arabidopsis seeds, a decline in its expression was noted but only after radicle emergence, i.e. after germination had been completed. Taken together, these data are consistent with GA-induced down-regulation of DELLA genes not being a prerequisite for germination of tomato, soybean, and Arabidopsis seeds. PMID- 15347802 TI - Molecular dimension explored in evolution to promote proteomic complexity. AB - The architecture of present-day protein interaction networks depends on how protein associations evolved. Here, we explore how and why evolution-related mutations influence protein structure to promote protein associations, and thereby network development. We specifically address two questions: (i) How can protein folds remain conserved while proteins accommodate new binding partnerships as genes duplicate? (ii) What is the structural/molecular basis for hub proteins being the most likely to acquire new connections? The answers stem from the examination of the structure wrapping, or protection from water attack. Wrapping is shown to be a crucial consideration in the exploration and evolution of proteomic interactivity. PMID- 15347803 TI - Improved peptide identification in proteomics by two consecutive stages of mass spectrometric fragmentation. AB - MS-based proteomics usually involves the fragmentation of tryptic peptides (tandem MS or MS(2)) and their identification by searching protein sequence databases. In ion trap instruments fragments can be further fragmented and analyzed, a process termed MS/MS/MS or MS(3). Here, we report that efficient ion capture in a linear ion trap leads to MS(3) acquisition times and spectra quality similar to those for MS(2) experiments with conventional 3D ion traps. Fragmentation of N- or C-terminal ions resulted in informative and low-background spectra, even at subfemtomol levels of peptide. Typically C-terminal ions are chosen for further fragmentation, and the MS(3) spectrum greatly constrains the C terminal amino acids of the peptide sequence. MS(3) spectra allow resolution of ambiguities in identification, a crucial problem in proteomics. Because of the sensitivity and rapid scan rates of the linear ion trap, several MS(3) spectra per peptide can be obtained even when sequencing very complex mixtures. We calculate the probability that an experimental MS(3) spectrum originates from fragmentation of a given N- or C-terminal ion of a peptide under consideration. This MS(3) identification score can be combined with the MS(2) scores of the precursor peptide from existing search engines. When MS(3) is performed on the linear ion trap-Fourier transform mass spectrometer combination, accurate peptide masses further increase confidence in peptide identification. PMID- 15347804 TI - Activating calcium-sensing receptor mutation in the mouse is associated with cataracts and ectopic calcification. AB - The extracellular calcium-sensing receptor (CaSR) plays a pivotal role in the regulation of extracellular calcium such that abnormalities, which result in a loss or gain of function, lead to hypercalcemia or hypocalcemia, respectively, in patients. Mice carrying CaSR knockout alleles develop hypercalcemia that mimics the disorders observed in humans. To date, there is no mouse model for an activating CaSR mutation. Here, we describe such a mouse model, named Nuf, originally identified for having opaque flecks in the nucleus of the lens in a screen for eye mutants. Nuf mice also display ectopic calcification, hypocalcemia, hyperphosphatemia, and inappropriately reduced levels of plasma parathyroid hormone. These features are similar to those observed in patients with autosomal dominant hypocalcemia. Inheritance studies of Nuf mice revealed that the trait was transmitted in an autosomal-dominant manner, and mapping studies located the locus to chromosome 16, in the vicinity of the CaSR gene (Mouse Genome Database symbol Gprc2a). DNA sequence analysis revealed the presence of a Gprc2a missense mutation, Leu723Gln. Transient expression of wild type and mutant CaSRs in human embryonic kidney 293 cells demonstrated that the mutation resulted in a gain of function of the CaSR, which had a significantly lower EC(50). Thus, our results have identified a mouse model for an activating CaSR mutation, and the development of ectopic calcification and cataract formation, which tended to be milder in the heterozygote Nuf mice, indicates that an evaluation for such abnormalities in autosomal dominant hypocalcemia patients who have activating CaSR mutations is required. PMID- 15347805 TI - Hyperleptinemia prevents lipotoxic cardiomyopathy in acyl CoA synthase transgenic mice. AB - The physiologic function of the progressive hyperleptinemia of diet-induced obesity is unknown. However, that lipotoxicity in nonadipose tissues of congenitally unleptinized obese rodents is far greater than in hyperleptinemic diet-induced obesity rodents has suggested an antilipotoxic role. To test this hypothesis, mice with severe lipotoxic cardiomyopathy, induced transgenically by cardiomyocyte-specific overexpression of the acyl CoA synthase (ACS) gene, were made hyperleptinemic by treatment with recombinant adenovirus containing the leptin cDNA. Normoleptinemic control ACS-transgenic mice developed severe dilated cardiomyopathy with thickened left ventricular walls and profound impairment of systolic function on echocardiogram; histologically, there was severe myofiber disorganization and interstitial fibrosis, with intracytoplasmic lipid vacuoles identifiable by electron microscope. By contrast, the hearts of hyperleptinemic ACS-transgenic mice appeared normal, with normal echocardiograms and cardiac triglyceride (TG) contents. Their lower myocardial TG content was ascribed primarily to profound lowering of plasma TG and free fatty acids; free fatty acids were 17% of normal at 8 weeks. Additionally, enhanced myocardial AMP activated protein kinase phosphorylation may have increased fatty acid oxidation, thereby contributing to the lowering of lipid stores. We conclude that obesity level hyperleptinemia protects the heart from lipotoxicity. PMID- 15347806 TI - Behavioral and regulatory abnormalities in mice deficient in the NPAS1 and NPAS3 transcription factors. AB - Laboratory mice bearing inactivating mutations in the genes encoding the NPAS1 and NPAS3 transcription factors have been shown to exhibit a spectrum of behavioral and neurochemical abnormalities. Behavioral abnormalities included diminished startle response, as measured by prepulse inhibition, and impaired social recognition. NPAS1/NPAS3-deficient mice also exhibited stereotypic darting behavior at weaning and increased locomotor activity. Immunohistochemical staining assays showed that the NPAS1 and NPAS3 proteins are expressed in inhibitory interneurons and that the viability and anatomical distribution of these neurons are unaffected by the absence of either transcription factor. Adult brain tissues from NPAS3- and NPAS1/NPAS3-deficient mice exhibited a distinct reduction in reelin, a large, secreted protein whose expression has been reported to be attenuated in the postmortem brain tissue of patients with schizophrenia. These observations raise the possibility that a regulatory program controlled in inhibitory interneurons by the NPAS1 and NPAS3 transcription factors may be either substantively or tangentially relevant to psychosis. PMID- 15347807 TI - A mucosally targeted subunit vaccine candidate eliciting HIV-1 transcytosis blocking Abs. AB - A vaccine that would engage the mucosal immune system against a broad range of HIV-1 subtypes and prevent epithelial transmission is highly desirable. Here we report fusing the mucosal targeting B subunit of cholera toxin to the conserved galactosylceramide-binding domain (including the ELDKWA-neutralizing epitope) of the HIV-1 gp41 envelope protein, which mediates the transcytosis of HIV-1 across the mucosal epithelia. Chimeric protein expressed in bacteria or plants assembled into oligomers that were capable of binding galactosyl-ceramide and G(M1) gangliosides. Mucosal (intranasal) administration in mice of the purified chimeric protein followed by an i.p. boost resulted in transcytosis-neutralizing serum IgG and mucosal IgA responses and induced immunological memory. Plant production of mucosally targeted immunogens could be particularly useful for immunization programs in developing countries, where desirable product traits include low cost of manufacture, heat stability, and needle-free delivery. PMID- 15347808 TI - A family of variably expressed outer-membrane proteins (Vomp) mediates adhesion and autoaggregation in Bartonella quintana. AB - Bartonella species are fastidious, Gram-negative human pathogens that can persist in the host bloodstream for years and bind to and invade several types of host cells. For many pathogens, adhesion to host cells and extracellular matrix (ECM) components is a critical virulence determinant. Bacteria often vary expression of surface adhesins by phase or antigenic variation to subvert the host immune response and permit adaptive interaction with different host structures. We developed a macaque animal model for Bartonella quintana infection to detect changes in bacterial outer-membrane proteins (OMP) during prolonged bloodstream infection. We identified a gene family encoding four highly conserved, 100-kDa, variably expressed OMP (Vomp), two of which function as adhesins. The variable expression of Vomp family members appears to be mediated by deletion of one or more vomp genes during chronic bloodstream infection. vomp deletion was observed also in isolates from humans with chronic B. quintana infection. The Vomp are closely related to the afimbrial adhesin, YadA, a virulence factor of Yersinia enterocolitica. The surface-expressed Vomp contain conserved structural features of YadA, including collagen-binding motifs. We demonstrate that the B. quintana Vomp are multifunctional OMP involved in binding to collagen and autoaggregation: VompC confers the ability to bind collagen IV, and VompA is necessary and sufficient for autoaggregation. The B. quintana Vomp are members of the newly recognized family of YadA-like trimeric autotransporters; the Vomp constitute a multigene family, they are variably expressed, and different virulence properties are attributable to individual Vomp family members. PMID- 15347809 TI - Identification of key phosphorylation sites in the circadian clock protein KaiC by crystallographic and mutagenetic analyses. AB - In cyanobacteria, KaiC is an essential hexameric clock protein that forms the core of a circadian protein complex. KaiC can be phosphorylated, and the ratio of phospho-KaiC to non-phospho-KaiC is correlated with circadian period. Structural analyses of KaiC crystals identify three potential phosphorylation sites within a 10-A radius of the ATP binding regions that are at the T432, S431, and T426 residues in the KaiCII domains. When these residues are mutated by alanine substitution singly or in combination, KaiC phosphorylation is altered, and circadian rhythmicity is abolished. These alanine substitutions do not prevent KaiC from hexamerizing. Intriguingly, the ability of KaiC overexpression to repress its own promoter is also not prevented by alanine substitutions at these sites, implying that the capability of KaiC to repress its promoter is not sufficient to allow the clockwork to oscillate. The KaiC structure and the mutational analysis suggest that S431 and T426 may share a phosphate that can shuttle between these two residues. Because the phosphorylation status of KaiC oscillates over the daily cycle, and KaiC phosphorylation is essential for clock function as shown here, daily modulations of KaiC activity by phosphorylation at T432 and S431/T426 seem to be key components of the circadian clockwork in cyanobacteria. PMID- 15347810 TI - Ten- to 50-nm-long quasi-ballistic carbon nanotube devices obtained without complex lithography. AB - A simple method combining photolithography and shadow (or angle) evaporation is developed to fabricate single-walled carbon nanotube (SWCNT) devices with tube lengths of approximately 10-50 nm between metal contacts. Large numbers of such short devices are obtained without the need of complex tools such as electron beam lithography. Metallic SWCNTs with lengths of approximately 10 nm, near the mean free path of l(op) approximately 15 nm for optical phonon scattering, exhibit nearly ballistic transport at high biases and can carry unprecedented 100 microA currents per tube. Semiconducting SWCNT field-effect transistors with approximately 50-nm channel lengths are routinely produced to achieve quasi ballistic operations for molecular transistors. The results demonstrate highly length-scaled and high-performance interconnects and transistors realized with SWCNTs. PMID- 15347811 TI - Fast folding of a helical protein initiated by the collision of unstructured chains. AB - To examine whether helix formation necessarily precedes chain collision, we have measured the folding of a fully helical coiled coil that has been specially engineered to have negligible intrinsic helical propensity but high overall stability. The folding rate approaches the diffusion-limited value and is much faster than possible if folding is contingent on precollision helix formation. Therefore, the collision of two unstructured chains is the initial step of the dominant kinetic pathway, whereas helicity exerts its influence only at a later step. Folding from an unstructured encounter complex may be efficient and robust, which has implications for any biological process that couples folding to binding. PMID- 15347812 TI - Role of KaiC phosphorylation in the circadian clock system of Synechococcus elongatus PCC 7942. AB - In the cyanobacterium Synechococcus elongatus PCC 7942, KaiA, KaiB, and KaiC are essential proteins for the generation of a circadian rhythm. KaiC is proposed as a negative regulator of the circadian expression of all genes in the genome, and its phosphorylation is regulated positively by KaiA and negatively by KaiB and shows a circadian rhythm in vivo. To study the functions of KaiC phosphorylation in the circadian clock system, we identified two autophosphorylation sites, Ser 431 and Thr-432, by using mass spectrometry (MS). We generated Synechococcus mutants in which these residues were substituted for alanine by using site directed mutagenesis. Phosphorylation of KaiC was reduced in the single mutants and was completely abolished in the double mutant, indicating that KaiC is also phosphorylated at these sites in vivo. These mutants lost circadian rhythm, indicating that phosphorylation at each of the two sites is essential for the control of the circadian oscillation. Although the nonphosphorylatable mutant KaiC was able to form a hexamer in vitro, it failed to form a clock protein complex with KaiA, KaiB, and SasA in the Synechococcus cells. When nonphosphorylatable KaiC was overexpressed, the kaiBC promoter activity was only transiently repressed. These results suggest that KaiC phosphorylation regulates its transcriptional repression activity by controlling its binding affinity for other clock proteins. PMID- 15347813 TI - Olfactory receptor surface expression is driven by association with the beta2 adrenergic receptor. AB - Olfactory receptors (ORs) comprise more than half of the large class I G protein coupled receptor (GPCR) superfamily. Although cloned over a decade ago, little is known about their properties because wild-type ORs do not efficiently reach the cell surface following heterologous expression. Receptor-receptor interactions strongly influence surface trafficking of other GPCRs, and we examined whether a similar mechanism might be involved in OR surface expression. Olfactory neurons are known to express beta-adrenergic receptors (ARs), and we found that coexpression with beta(2)-ARs, but not any other AR subtypes, dramatically increased mouse 71 (M71) OR surface expression in human embryonic kidney 293 cells. A persistent physical interaction between M71 ORs and beta(2)-ARs was shown by coimmunoprecipitation and by cointernalization of the two receptors in response to their specific ligands. Also, coexpression of wild-type M71 ORs with beta(2)-ARs resulted in cAMP responses to the M71 ligand acetophenone. Finally, in situ hybridization studies showed extensive colocalization of M71 OR and beta(2)-AR expression in mouse olfactory epithelium. These data demonstrate the successful heterologous surface expression of a functional wild-type OR and reveal that persistent physical association with other GPCRs can control OR surface expression. PMID- 15347814 TI - Distinct roles of transcription factors TFIIIB and TFIIIC in RNA polymerase III transcription reinitiation. AB - Eukaryotic RNA polymerase (Pol) III is recruited to target promoters by a stable preinitiation complex containing transcription factors TFIIIC and TFIIIB. After the first transcription cycle, reinitiation proceeds through facilitated recycling, a process by which the terminating Pol III rapidly reloads onto the same transcription unit. Here, we show that Pol III is repeatedly recaptured in vitro by the first transcribed gene, even in the presence of a juxtaposed competitor promoter complex, thus suggesting that facilitated recycling is not merely due to a stochastic reassociation process favored by the small size of class III genes. The transcription factor requirements for facilitated reinitiation were investigated by taking advantage of Pol III templates that support both TFIIIC-dependent and TFIIIC-independent transcription. A TFIIIC-less transcription system, in which TFIIIB was reconstituted from recombinant TATA box binding protein and Brf1 proteins and a crude fraction containing the Bdp1 component, was sufficient to direct efficient Pol III recycling on short ( approximately 100 bp) class III genes. Unexpectedly, however, on longer (>300 bp) transcription units, reinitiation in the presence of TFIIIB alone was compromised, and TFIIIC was further required to reestablish a high reinitiation rate. Transcription reinitiation was also severely impaired when recombinant Bdp1 protein replaced the corresponding crude fraction in reconstituted TFIIIB. The data reveal an unexpected complexity in the Pol III reinitiation mechanism and suggest the existence of a handing-back network between Pol III, TFIIIC, and TFIIIB on actively transcribed class III genes. PMID- 15347815 TI - Phylogenetic discovery bias in Bacillus anthracis using single-nucleotide polymorphisms from whole-genome sequencing. AB - Phylogenetic reconstruction using molecular data is often subject to homoplasy, leading to inaccurate conclusions about phylogenetic relationships among operational taxonomic units. Compared with other molecular markers, single nucleotide polymorphisms (SNPs) exhibit extremely low mutation rates, making them rare in recently emerged pathogens, but they are less prone to homoplasy and thus extremely valuable for phylogenetic analyses. Despite their phylogenetic potential, ascertainment bias occurs when SNP characters are discovered through biased taxonomic sampling; by using whole-genome comparisons of five diverse strains of Bacillus anthracis to facilitate SNP discovery, we show that only polymorphisms lying along the evolutionary pathway between reference strains will be observed. We illustrate this in theoretical and simulated data sets in which complex phylogenetic topologies are reduced to linear evolutionary models. Using a set of 990 SNP markers, we also show how divergent branches in our topologies collapse to single points but provide accurate information on internodal distances and points of origin for ancestral clades. These data allowed us to determine the ancestral root of B. anthracis, showing that it lies closer to a newly described "C" branch than to either of two previously described "A" or "B" branches. In addition, subclade rooting of the C branch revealed unequal evolutionary rates that seem to be correlated with ecological parameters and strain attributes. Our use of nonhomoplastic whole-genome SNP characters allows branch points and clade membership to be estimated with great precision, providing greater insight into epidemiological, ecological, and forensic questions. PMID- 15347816 TI - Resolution of the insect ouabain paradox. AB - Many insects are highly resistant to plant toxins, such as the cardiac glycoside ouabain. How can the epithelia that must handle such toxins, also be refractory to them? In Drosophila, the Malpighian (renal) tubule contains large amounts of Na(+),K(+) ATPase that is known biochemically to be exquisitely sensitive to ouabain, yet the intact tissue is almost unaffected by even extraordinary concentrations. The explanation is that the tubules are protected by an active ouabain transport system, colocated with the Na(+),K(+) ATPase, thus preventing ouabain from reaching inhibitory concentrations within the basolateral infoldings of principal cells. These data show that the Na(+),K(+) ATPase, previously thought to be unimportant, may be as vital in insect tissues as in vertebrates, but can be cryptic to conventional pharmacology. PMID- 15347817 TI - Reduction of nitrite to nitric oxide during ischemia protects against myocardial ischemia-reperfusion damage. AB - Nitric oxide (NO.) is thought to protect against the damaging effects of myocardial ischemia-reperfusion injury, whereas xanthine oxidoreductase (XOR) normally causes damage through the generation of reactive oxygen species. In the heart, inorganic nitrite (NO(2)(-)) has the potential to act as an endogenous store of NO., liberated specifically during ischemia. Using a detection method that we developed, we report that under ischemic conditions both rat and human homogenized myocardium and the isolated perfused rat heart (Langendorff preparation) generate NO. from NO(2)(-) in a reaction that depends on XOR activity. Functional studies of rat hearts in the Langendorff apparatus showed that nitrite (10 and 100 microM) reduced infarct size from 47.3 +/- 2.8% (mean percent of control +/- SEM) to 17.9 +/- 4.2% and 17.4 +/- 1.0%, respectively (P < 0.001), and was associated with comparable improvements in recovery of left ventricular function. This protective effect was completely blocked by the NO. scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazole-1-oxyl 3-oxide (carboxy-PTIO). In summary, the generation of NO. from NO(2)(-), by XOR, protects the myocardium from ischemia-reperfusion injury. Hence, if XOR is presented with NO(2)(-) as an alternative substrate, the resultant effects of its activity may be protective, by means of its production of NO. , rather than damaging. PMID- 15347818 TI - A multistep approach in the cytologic evaluation of liver biopsy samples of dogs with hepatic diseases. AB - Cytologic criteria were evaluated for their diagnostic value in liver disease in dogs. Therefore, histopathologic and cytologic examination was performed on liver biopsy samples of 73 dogs with liver diseases and 28 healthy dogs. Logistic regression analysis was used to select the measured parameters to be included in a multistep approach. With the logistic regression method, different characteristic cytologic parameters could be defined for each histopathologic diagnosis. In malignant lymphoma of the liver, the presence of large numbers of lymphoblasts with a minimum of 5% of all cells was found. Clusters of epithelial cells with several cytologic characteristics of malignancy intermixed with normal hepatocytes were indicative of metastatic carcinoma or cholangiocellular carcinoma. Liver cells in hepatocellular carcinoma were characterized by a high nucleus/cytoplasm ratio, large cell diameters, increased numbers of nucleoli per nuclei, small numbers of cytoplasmic vacuoles, and frequently, small numbers of lymphocytes. Extrahepatic cholestasis was characterized by excessive extracellular bile pigment in the form of biliary casts, an increased number of nucleoli within hepatocytes, decreased hepatic cell size, and low numbers of lymphocytes. In destructive cholangiolitis, increased numbers of neutrophils and a small mean nuclear size within hepatocytes were seen. Acute and nonspecific reactive hepatitis are diagnosed based on the presence of moderate reactive nuclear patterns, including more pronounced chromatin, prominent nucleoli, increased numbers of inflammatory cells, excluding lymphocytes, and the absence of increased numbers of bile duct cell clusters. Increased number of mast cells also was indicative of nonspecific reactive hepatitis. Important cytologic criteria for the diagnosis of liver cirrhosis, in addition to chronic hepatitis, are intracellular bile accumulation and increased numbers of bile duct cell clusters. In summary, the stepwise approach based on logistic regression presented in this study might be helpful in the objective cytologic diagnosis of hepatic diseases. PMID- 15347819 TI - Features of cell degeneration and death in hepatic failure and systemic lymphoid depletion characteristic of porcine circovirus-2-associated postweaning multisystemic wasting disease. AB - Tissue section replicates from lymphoid tissues and livers of gnotobiotic swine were examined by immunohistochemistry for the colocalization of porcine circovirus-2 (PCV-2) nucleocapsid and terminal deoxynucleotidyl transferase (TdT) mediated incorporation of biotinylated nucleotides (UTP) onto the 3'-exposed hydroxyl groups (nick end labeling) nuclear deoxyribonucleic acid (TUNEL), a marker for apoptosis. Single- and dually stained replicates from uninfected controls, subclinically affected PCV-2-infected gnotobiotic pigs, PCV-2-infected piglets immunosuppressed with cyclosporine (Cys), and PCV-2-infected piglets with post-weaning multisystemic wasting syndrome (PMWS) were evaluated. Thymuses were used as positive controls for apoptosis absent PCV-2, tissue sections from dogs given hyperthermic stress were examined as positive controls for induced TUNEL. Tissues from heat-stressed dogs contained TUNEL-positive cell nuclei in both lymphoid tissues and liver, TUNEL was greatest shortly after the delivery of the hyperthermic insult. In uninfected control and subclinically affected PCV-2 infected gnotobiotic pigs, rare hepatocytes and lymphoid cells were TUNEL positive, the frequency of these was similar to that seen in uninfected controls. In PMWS-affected and Cys-treated PCV-2 piglets, the only consistent strongly positive TUNEL signal was contained within the cytoplasm of virus-positive phagocytic mononuclear cells. In phagocytes, some PCV-2 inclusions were TUNEL positive. Collectively, these data indicate that apoptosis is not the primary mechanism of lymphoid depletion and hepatocyte loss in PMWS. Apoptosis associated with systemic viral diseases may be attributable to pyrexia rather than direct or indirect effects of viruses on target cells. PMID- 15347820 TI - Pathogenicity by parenteral injection of fowl adenovirus isolated from gizzard erosion and resistance to reinfection in adenoviral gizzard erosion in chickens. AB - The pathogenicity of a serotype-1 fowl adenovirus (FAV-99ZH), which causes adenoviral gizzard erosion by oral inoculation in chickens, was investigated in specific pathogen-free white leghorn chickens. In trial 1, 14 chickens were inoculated intravenously with the virus at 21 days of age and euthanatized for necropsy within 1-14 days of inoculation. Gizzard erosion was grossly observed from day 7 postinoculation (PI), and histologically, FAV-99ZH antigen-positive, basophilic intranuclear inclusion bodies were seen in the gizzard lesions from day 7 to 11 PI. Necrotizing pancreatitis, and cholecystitis and cholangitis associated with the inclusions were observed from day 3 to 14 PI (pancreatitis) and from day 5 to 9 PI (cholecystitis and cholangitis), respectively. The inclusions were also observed in the epithelial cells of the cecal tonsils from day 3 to 5 PI. The virus was recovered from samples of the lesions. It was revealed that FAV-99ZH causes not only gizzard erosion but also pancreatitis, cholecystitis, and cholangitis by intravenous inoculation in chickens. In trial 2, 10 chickens were inoculated orally with the virus twice, at 13 and 36 days of age, and euthanatized for necropsy within 4-17 days after reinfection. Macroscopically, focal gizzard lesions were observed; however, neither necrosis nor inclusions were observed by microscopy. Moreover, FAV was not recovered from the gizzard or rectum of any of the chickens at necropsy. This suggests that the gizzard lesions occurred as a result of the primary infection, and that the chickens were able to resist reinfection. PMID- 15347821 TI - Indicators of malignancy of canine adrenocortical tumors: histopathology and proliferation index. AB - Tumors of the adrenal cortex account for 10-20% of the naturally occurring Cushing's syndrome diagnosed in dogs. Differentiating between adrenocortical adenoma and carcinomas is often difficult. The purposes of this study were to determine which histopathologic criteria can be used as markers for malignancy in canine adrenocortical tumors and the relevance of the proliferation marker, Ki 67, for differentiation between cortical adenomas and carcinomas. Twenty-six adrenocortical carcinomas, 23 adenomas, and 11 normal adrenal glands were examined. Morphologic criteria significantly associated with adrenocortical carcinomas included a size larger than 2 cm in diameter, peripheral fibrosis, capsular invasion, trabecular growth pattern, hemorrhage, necrosis, and single cell necrosis, whereas hematopoiesis, fibrin thombi, and cytoplasmic vacuolation were significantly associated with adrenocortical adenomas. The mean (+/- SD) proliferation index, measured by immunohistochemistry for the Ki-67 antigen, was 9.3 +/- 6.3% in carcinomas, 0.76 +/- 0.83% in adenomas, and 0.58 +/- 0.57% in normal adrenal glands. The Ki-67 proliferation index was significantly higher in carcinomas compared with adenomas and normal adrenal glands. A threshold value of the proliferation index of 2.4% reliably separated carcinomas from adenomas. Based on these results, it appears that thorough evaluation of morphologic features combined with immunohistochemical assessment of the proliferation index is extremely useful for differentiating between adrenocortical adenomas and carcinomas in dogs. PMID- 15347822 TI - Kinetics of infection and effects on the placenta of Chlamydophila abortus in experimentally infected pregnant ewes. AB - A Chlamydophila abortus-induced abortion model was carried out on the basis of the experimental infection of ewes at day 75 of gestation. The infection induced abortions and the birth of weak lambs during the last 3 weeks of pregnancy. To study the kinetics of the infection in the placenta and in other organs, infected ewes were killed at 105, 120, and 130 days of gestation and also several days after abortion or parturition. Infected ewes developed a systemic infection that caused a mild and transient pneumonia and focal hepatitis. Pathologic changes were observed in placentas at 120 day of gestation, although the lesions varied between animals and even between placentomes of the same placenta. The first placental area infected was the maternal stroma and epithelium next to the intercaruncular areas, where neutrophilic response seemed to control the infection. A substantial degree of multiplication of C. abortus was then observed in the trophoblast cells of the placentome, periplacentomal choriallantoic membranes, and hilius, with an inflammatory exudate composed mainly of neutrophils, some macrophages, and very scarce lymphocytes. After abortion, the lesions affected the intercotyledonary areas of the aborted placentas, whereas in the uterus significant lymphocyte infiltration was observed, together with a rapid decrease of the C. abortus antigen in the degenerated caruncular tissues. PMID- 15347823 TI - Disseminated infection with Balamuthia mandrillaris in a dog. AB - Bilateral chronic granulomatous nephritis and meningoencephalitis were diagnosed on necropsy of a 2-year-old male Great Dane dog. The causative agent was identified as Balamuthia mandrillaris, based on morphologic features, immunohistochemical staining, and deoxyribonucleic acid detection using the polymerase chain reaction with newly designed primer pairs. Trophozoite and cystic forms of the amoeba were evident within the kidneys and brain parenchyma. This is the first report on a B. mandrillaris infection in a dog. PMID- 15347824 TI - Intracranial fusariosis: a novel cause of fungal meningoencephalitis in a dog. AB - The Fusarium species are a group of saprophytic fungal organisms that are occasionally the cause of opportunistic infections in humans and animals. Central nervous system disease associated with a Fusarium species is most commonly described in horse, resulting in equine leukoencephalomalacia. This report describes a 2-year-old, spayed, female German Shepherd Dog with meningoencephalitis secondary to infection with Fusarium spp. Meningoencephalitis in dogs secondary to a species of Fusarium has not been previously reported. The diagnosis was made based on the histopathologic examination of brain tissues postmortem and special immunohistochemical stains specific for Fusarium solani. The clinical signs in this dog were indicative of multifocal brain disease and included seizures and a paradoxical vestibular syndrome. The clinical findings, diagnostic and histopathologic test results, and the comparative characterizations of other disseminated fungal diseases, especially aspergillosis, are described. PMID- 15347825 TI - Type A-like retroviral particles in a metastatic intestinal adenocarcinoma in an emerald tree boa (Corallus caninus). AB - A metastatic intestinal papillary adenocarcinoma was diagnosed histologically in an emerald tree boa (Corallus caninus). Metastasis was detected in the liver, both kidneys, lung, and coelomic wall. Ultrastructural examination of the metastatic intestinal epithelial cells in the liver revealed the presence of a moderate number of viral particles that most closely resembled type A retroviral particles and were mainly associated with granular endoplasmic reticulum membranes. This case is the first description of type A-like retroviral particles in a neoplasm of a snake. The role of the virions in the etiology of the intestinal adenocarcinoma is uncertain. In addition, this is the first confirmed report of a metastatic intestinal adenocarcinoma in a snake. PMID- 15347826 TI - Congenital fetal rhabdomyoma in a foal. AB - An Appaloosa filly was born with a ventral midline, approximately 8 x 12 x 15 cm subcutaneous cervical mass. The nonencapsulated mass was composed of interlacing and haphazard bundles of spindle cells on moderate to abundant loose myxomatous stroma. A moderate number of cells showed cross striations with minor nuclear variation and a low mitotic rate. Immunohistochemical staining for myoglobin, desmin, actin, vimentin, and S-100 was positive and negative for glial fibrillar antigen and keratin. Rhabdomyomas are rare benign tumors of striated muscle. Rhabdomyomas described previously in the veterinary literature are analogous to the "adult form" of human rhabdomyoma. This is the first report of a veterinary case that 1) clinically and histologically parallels the "fetal form" in human rhabdomyoma and 2) describes a congenital extracardiac rhabdomyoma. PMID- 15347827 TI - Pulmonary neoplasia in two llamas (Lama glama). AB - Two llamas with pulmonary tumors were examined. Llama No. 1 had multiple nodules throughout the lung that consisted histologically of solid clusters of polygonal to spindle cells with rare glandular differentiation. Intravascular emboli were common. Similar neoplastic masses were present in the kidney, heart, and liver. Immunohistochemically, neoplastic cells were positive for broad-spectrum cytokeratins (CKs), high-molecular weight CKs, CKs 5/6, and vimentin. The diagnosis was pulmonary carcinoma. Llama No. 2 had pulmonary nodules without extrapulmonary involvement. Microscopically, neoplastic cells formed acini lined by simple epithelium and solid cords of squamous cells that sometimes surrounded acini. Neoplastic cells were strongly positive for broad-spectrum CKs and weakly positive for thyroid transcription factor-1. The diagnosis was adenosquamous carcinoma. Pulmonary tumors account for 23% of neoplasms in South American camelids in our laboratory, making this the second most common type of neoplasm after lymphosarcoma. PMID- 15347828 TI - Oligodendroglioma in the cervical spinal cord of a dog. AB - A 7-year-old male castrated Yorkshire Terrier dog developed slowly progressive neurologic disturbances consisting of difficulties in moving the neck, lack of proprioception, and tetraparesis 4 months prior its death. Neurologic examination, computer tomography, and myelography resulted in the tentative diagnosis of intramedullary cervicothoracic spinal cord lesion. At necropsy, an intramedullary cervical spinal cord mass between C5 and C6 was noticed. Histologically, cells of this well-demarcated, nonencapsulated neoplasm were arranged in sheaths or cords separated by a fine fibrovascular stroma. The polygonal to round tumor cells were characterized by moderate pale, basophilic, and vacuolar cytoplasm and round to slightly oval, centrally located nuclei with fine-stippled heterochromatin, a single nucleolus, and a very low mitotic activity. Tumor cells lacked glial fibrillary acidic protein, vimentin, factor VIII-related, and cytokeratin antigen expression. Histologic and immunohistochemical findings led to the diagnosis of a cervical spinal cord oligodendroglioma. PMID- 15347829 TI - Leukoencephalitis associated with selective viral replication in the brain of a pony with experimental chronic equine infectious anemia virus infection. AB - Neurologic disease occurs sporadically in horses infected with the equine infectious anemia virus (EIAV). This report describes a case of clinically severe neurologic disease in a pony experimentally infected with EIAV. This pony did not have fever or anemia, which are the characteristic clinical signs of disease. The histopathologic changes were characterized as lymphohistiocytic periventricular leukoencephalitis. Polymerase chain reaction and in situ hybridization data showed that the brain lesions were directly associated with viral replication and that high-level viral replication occurred selectively within the lesion and not in other tissues. These findings suggest that EIAV-associated neurologic disease is the direct result of viral replication. PMID- 15347830 TI - Multifocal pleural cystic squamous metaplasia in a horse with chronic obstructive bronchopneumonia. AB - A 10-year-old Knabstrupper stallion was euthanatized because of severe dyspnea and exercise intolerance. Postmortem examination revealed diffuse severe alveolar emphysema and chronic fibrosing pleuritis of the caudal lung. Parts of both caudal lung lobes were covered with multiple raised firm gray to yellow plaques. Histologically, these areas consisted of circumscribed pleural fibroses and cysts of metaplastic keratinizing squamous epithelium. Immunohistochemistry revealed intense labeling for cytokeratins 5/6 and 10. In addition, caudal lung lobes were severely affected by a chronic partially obliterative bronchiolitis and peribronchiolitis with multifocal pleural involvement. PMID- 15347834 TI - Enzyme-to-enzyme channeling in the early steps of glycolysis in rat pancreatic islets. AB - The enzyme-to-enzyme channeling of metabolic intermediates is not an uncommon process. The present review draws attention to recent experimental work documenting, in rat pancreatic islets, the enzyme-to-enzyme channeling of alpha-D glucose 6-phosphate between hexokinase isoenzyme(s), mainly glucokinase, and phosphoglucoisomerase. Likewise, the possible enzyme-to-enzyme channeling of beta D-fructose 6-phosphate between phosphoglucoisomerase and phosphofructokinase is briefly evoked. These considerations are relevant to the anomeric specificity of D-glucose metabolism, even in islets exposed to equilibrated D-glucose, to the perturbation of such an anomeric specificity in the phenomenon of so-called B cell glucotoxicity, and to the correct interpretation of 3HOH generation from D [2-(3H)]glucose. PMID- 15347835 TI - The HFE gene is associated to an earlier age of onset and to the presence of diabetic nephropathy in diabetes mellitus type 2. AB - We initiated the present work to determine whether the presence of the HFE C282Y or H63D mutations could be related to the clinical expression of diabetes mellitus type 2. Two hundred and twenty five type 2 consecutive diabetic patients were included and the HFE genotypes were determined. Younger ages of onset of diabetes as well as a longer duration of the disease were detected in patients carrying at least one C282Y allele (p = 0.007). An increased prevalence of retinopathy (p = 0.014) and of nephropathy (p = 0.04) were also detected in individuals carrying at least one C282Y allele in comparison with patients carrying the other alleles. The increased prevalence of retinopathy in C282Y carriers is related to the increased duration of the disease, but we not have detected that the prevalence of nephropathy is associated with the duration of the disease. However, multivariate logistic regression confirms that the prevalence of nephropathy is higher in the group of patients carrying at least one C282Y allele or the H63D/H63D genotype as compared to the group of patients with the wild-type (N/N) or the N/H63D genotype. To our knowledge our study is the first one to report an earlier age of onset in type 2 diabetic patients carrying HFE mutations. PMID- 15347836 TI - Immunization against endogenous inhibin increases normal oocyte/embryo production in adult mice. AB - The objectives of the present study were to induce super-ovulation in adult mice by immunoneutralization of endogenous inhibin and to investigate embryo development in vivo and in vitro. Adult female mice of the ddY strain at 3 mo of age were superovulated with a single ip injection of inhibin antiserum (inhibin AS; 50, 100, or 200 micro L) at 12:00 h on metestrus of the 4-d estrous cycle. The control group was treated with 100 micro L normal goat serum (control serum). Five mice were sacrificed every 6 h and blood samples were collected for hormonal assay. Individual female mice were bred in the afternoon of proestrus with individual fertile males. Embryos were collected at 08.00 h on d 2 post-coitus and cultured in KSOM solution. Treatment with inhibin-AS significantly increased the concentrations of plasma FSH. Furthermore, there was a significant elevation in plasma concentrations of estradiol-17beta and progesterone in the inhibin immunized groups in comparison with controls. The superovulated oocytes that were fertilized normally in vivo were able to develop normally to blastocysts in vitro. The number of oocytes and blastocysts developed in animals treated with inhibin-AS was significantly higher than that for mice treated with control serum. Moreover, the rate of fertilization and the rate of blastocyst development were similar in inhibin-AS treated and control groups. These results indicate that immunoneutralization of endogenous inhibin induces superovulation in adult mice without additional treatment with human chorionic gonadotropin (hCG). In addition, the superovulated oocytes obtained by administration of inhibin-AS have normal embryonic developmental competence. PMID- 15347837 TI - Atorvastatin enhances bone density in ovariectomized rats given 17beta-estradiol or human parathyroid hormone(1-34). AB - We investigated the in vivo effect of atorvastatin on bone mineral density (BMD) in ovariectomized (OVX) rats. Eight-week-old female rats underwent either a sham operation or ovariectomy, and treatments with vehicle, atorvastatin, 17beta estradiol (E2) and human parathyroid hormone(1-34) [hPTH(1-34)] were initiated 6 wk after the surgery. E2 (10 microg/kg) treatment for 12 wk significantly increased lumbar BMD (L2-L4), whereas atorvastatin did not affect lumbar BMD. The combination of atorvastatin (2 mg/kg) and E2 significantly enhanced the BMD of lumbar vertebrae (L2-L4) and femoral metaphyseal area (2/10-4/10 segments from the most proximal point) compared to that of either E2 or atorvastatin alone. A low dose 1 micro g/kg of hPTH (1-34) did not alter lumbar or femoral BMD, whereas a high dose 17.5 micro g/kg of the peptide significantly increased BMD. Concomitant injections of atorvastatin (2 mg/kg) with hPTH(1-34) (1 microg/kg) for 8 wk significantly enhanced the BMD of lumbar vertebrae and the metaphyseal area of the femur in OVX rats. These findings demonstrate that chronic administration of atorvastatin appears to modestly enhance the BMD of the lumbar vertebrae and femoral metaphysis of OVX rats treated with submaximal doses of E2 and hPTH(1-34). PMID- 15347838 TI - Heparin increases prolactin and modifies the effects of fgf-2 upon prolactin accumulation in pituitary primary cultures. AB - We have studied the effects of heparin on prolactin accumulation in the medium from primary pituitary cultures, and whether heparin interferes with the effects of fibroblast growth factor-2 (FGF-2) on PRL regulation in vitro. In the absence of exogenous FGF-2, hepa-rin increased prolactin accumulation in the culture medium in a dose-dependent manner. FGF-2 also increased the prolactin levels of primary cells in a time- and dose-dependent manner. However, low doses of heparin reduced the effects of FGF-2, but higher doses of heparin increased the maximal FGF-2-induced prolactin secretion and ED50. In vivo estrogenization of rats resulted in the abolition of FGF-2 capability to promote prolactin release in vitro. However, heparin restored cell responsiveness to FGF-2. Our results suggest that heparin, when present in the medium, binds FGF-2, therefore reducing its ability to interact with FGF receptors in a dose-dependent manner up to a critical molar concentration, at which heparin itself starts to activate the FGF receptor, and strengthens the activation induced by its proper ligand, FGF-2. Prolactin responses to FGF-2 are blocked by estrogen pretreatment, and it is probable that this introduces lactotroph cells in the proliferative stage. In conclusion, heparin modulates PRL secretion and PRL responses to FGF-2 in vitro. PMID- 15347839 TI - Gonadectomy alters myosin heavy chain composition in isolated cardiac myocytes. AB - Sex differences in cardiac function have been identified. Studies suggest that the presence of testosterone in males may contribute to the observed differences in cardiac function. Our laboratory has shown previously that testosterone treatment of gonadectomized adult male rats enhances contractility of isolated rat ventricular myocytes. In this study we tested the hypothesis that gonadectomy and hormone replacement influences contractility by altering myosin heavy chain (MHC) composition. To test this hypothesis we analyzed myosin isoform expression in ventricular myocytes isolated from castrated rats displaying a decrease in myocyte contractile velocity and compared them to castrates treated with testosterone that displayed normal myocyte shortening velocity. Sixteen weeks after castration isolated rat ventricular myocytes displayed a 90% (p < 0.001) decline in MHC-alpha mRNA levels and over a twofold (p < 0.01) increase in MHC beta transcripts when compared to sham-operated controls. Consistent with these changes we also observed a substantial decline in the ratio of MHC-alpha to MHC beta protein expression. A reversal in myosin heavy chain composition was achieved following testosterone replacement. These studies provide the first direct evidence that testosterone replacement in gonadectomized animals enhances contractility via transcriptional and translational control of myosin heavy chain composition in isolated rat ventricular myocytes. The influence of testosterone on MHC composition in males may underlie some of the observed sex differences in cardiac function. PMID- 15347840 TI - Regulation of VEGF-A, VEGFR-I, thrombospondin-1, -2, and -3 expression in a human pituitary cell line (HP75) by TGFbeta1, bFGF, and EGF. AB - Pituitary tumors are highly vascular neoplasms, which suggest an important role of angiogenesis in pituitary tumor growth. We used the human pituitary cell line (HP75) to examine the effects of the growth factors TGFbeta1, bFGF, and EGF on cell growth, and on the regulation of the pro-angiogenic growth factor VEGF-A and the VEGFR-I and the anti-angiogenic molecules thrombospondin (TSP) TSP-1 and TSP 2 along with TSP-3. Real-time RT-PCR was used to measure mRNA levels, and Western blot was used to analyze TSP-1 and TSP-2 protein levels. TGFbeta1 treatment (1 x 10(-9) M) increased VEGF-A mRNA levels significantly (p < 0.05) after 4 and 24 h of treatment. TGF beta1 treatment decreased VEGF-R mRNA levels after 96 h of treatment (p < 0.05). After 96 h of treatment, TSP-1 and TSP-2 mRNA levels were significantly increased (p < 0.05) by TGFbeta1 treatment, which also inhibited HP75 cell growth. Basic FGF also increased TSP-1 mRNA levels after 96 h of treatment, but did not regulate growth of the pituitary tumor cells. Basic FGF and EGF did not modulate changes in VEGF-A mRNA levels after 4 and 24 h of treatment, but EGF increased VEGF-A significantly (p < 0.05) after 96 h of treatment. These results indicate that TGFbeta1 treatment may regulate angiogenesis in pituitary cells by initially increasing levels of pro-angiogenic VEGF-A and then stimulating the anti-angiogenic molecules TSP-1 and TSP-2 levels. PMID- 15347841 TI - Leptin, thyrotropin, and thyroid hormones in obese/overweight women before and after two levels of energy deficit. AB - The aim of our study was to compare serum concentration of leptin and pituitary thyroid axis hormones in obese/overweight women before and after two levels of energy deficit with those parameters in lean women on adequate energy intake. Additionally, we attempted to elucidate if the effect of weight reduction could be related to anthropometric and hormonal parameters before treatment. Anthropometric and hormonal parameters-serum leptin, TSH, T4, fT4, T3 and leptin to fat mass (Lep/fm), T3/T4, fT4/T4, T4/TSH, fT4/TSH--were compared in two groups of women (n =18 each)--lean women (C: BMI 22.0 +/- 1.2) and overweight/obese (Ov/Ob: BMI 29.9 +/- 3.3). Ov/Ob women were subjected to weight-reducing treatment consisting of energy intake equal to 80% of calculated total energy expenditure for the first 4 wk and to 50% for subsequent 4 wk. All baseline hormone concentrations, Lep/fm, and fT4/T4 were higher in overweight/obese group. After 20% energy deficit decrease in BMI, percent body fat (fm%), leptin, T3, and TSH serum concentrations as well as in Lep/fm and T3/T4 was observed; T4/TSH increased, fT4, fT4/T4 and fT4/TSH did not change significantly. Increase in energy deficit from 20% to 50% resulted in normalization of Lep/fm, on the other hand, it provoked greater decline in thyroid hormone plasma concentration, which could hinder further mass reduction. Leptin and TSH levels were positively correlated after 50% energy deficit treatment. Changes in fm% were directly related to baseline T4/TSH, fT4/TSH, and log TSH. In conclusion, TSH serum concentration and its ratio to T4 and fT4 before weight reduction could be a good predictor of successful weight loss. PMID- 15347842 TI - Serotonin may alter the pattern of gonadotropin-induced progesterone release of human granulosa cells in superfusion system. AB - Serotonin plays a hormonal function in several nonneuronal peripheral tissues, such as the ovaries. Our aim was to investigate whether there is a modulatory action of serotonin on gonadotropin-induced steroid secretion of human granulosa cells. In granulosa cell culture, serotonin was administered alone or in combination with luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Also, granulosa cells were transferred into a dynamic superfusion apparatus and challenged by FSH and LH alone or along with serotonin. Estradiol and progesterone concentrations of samples were measured by radioimmunoassay. As expected, administration of FSH, LH, and serotonin alone resulted in a significant estradiol and progesterone release in cell culture, as well as a significant increase in progesterone release in dynamic superfusion system. In cell culture, co-administration of serotonin with gonadotropins had no additive effect on gonadotropin-induced secretion of progesterone, while it further augmented that of estradiol. In superfusion system, when gonadotropins were added along with serotonin, the increase in progesterone release was markedly less, while peaks of hormone response were remarkably prolonged compared to challenges by LH and FSH alone. The observed effects of serotonin on gonadotropin-induced steroid release of granulosa cells may reveal further details about the regulation of granulosa cell function. PMID- 15347843 TI - Vascular responses to Angiotensin-(1-7) during the estrous cycle. AB - Mesenteric arteries (230-290 microm) were isolated from virgin female rats at diestrous and proestrous phases of the estrous cycle and from ovariectomized (OVX) rats with or without estrogen (E2) replacement (17beta-estradiol, 7.5 + 5 mg pellets, 21 d release). Arteries were mounted in a pressurized myograph system. Angiotensin-(1-7) [Ang-(1-7)] concentration-dependent responses (10(-10) 10(-5) M) were determined in arteries preconstricted with endothelin-1 (10(-7) M). Mesenteric arteries were pretreated with the specific Ang-(1-7) antagonist, D [Ala7]-Ang-(1-7) (10(-7) M) to assess the Ang-(1-7) receptor-mediated dilator effect. Ang-(1-7) did not dilate mesenteric arteries from virgin rats at diestrus and placebo-treated OVX female rats as compared to the time control; however, Ang (1-7) elicited a modest dilation at proestrus as compared to diestrus, which reached statistical significance at 10(-8) M concentrations. Ang-(1-7) caused a concentration-dependent vasodilation in mesenteric arteries of females with E2 replacement, with an EC50 of 21 nM. D-[Ala7]-Ang-(1-7) blocked the vasodilator effect of Ang-(1-7). Our results demonstrate that during proestrus Ang-(1-7) elicits modest vasodilation as compared to diestrus, but lacks vasodilatory properties in vessels from diestrous and ovariectomized rats. Estrogen replacement restores a significant dilator response to Ang-(1-7) in OVX rats that is mediated by a D-[Ala7]-Ang-(1-7) sensitive site. PMID- 15347844 TI - Nature of changes in adrenocortical function in chronic hyperleptinemic female rats. AB - Neonatal treatment of rats with monosodium L-glutamate, which destroys hypothalamic arcuate nucleus neuronal bodies, induces several metabolic abnormalities; as a result, rats develop a phenotype of pseudoobesity. This study was designed to explore, in the monosodium L-glutamate-treated female rat, the influence of chronic hyperleptinemia on adrenal cortex functionality. For this purpose, we evaluated in control and hypothalamic-damaged rats: (a) in vivo and in vitro adrenocortical function, (b) adrenal leptin receptor immunodistribution and mRNA expression, and (c) whether the inhibitory effect of leptin on adrenal function remains. Our results indicate that, compared to normal counterparts, pseudoobese animals displayed (1) hyperadiposity, despite being hypophagic and of lower body weight, (2) in vivo and in vitro enhanced adrenocortical response to ACTH stimulation, (3) an in vitro adrenal fasciculata-reticularis cell hyper sensitivity to ACTH stimulus, (4) hyperplasia of their adrenal zona fasciculata cells, and (5) adrenal fasciculata-reticularis cell refractoriness to the inhibitory effect of leptin on ACTH-stimulated glucocorticoid production due, at least in part, to decreased adrenal leptin receptor expression. These data further support that increased hypothalamo-pituitary-adrenal axis function, in the adult neurotoxin-lesioned female rat, is mainly dependent on the development of both hyperplasia of adrenal zona fasciculata and adrenal gland refractoriness to leptin inhibitory effect. Our study supports that adrenal leptin resistance could be responsible, at least in part, for enhanced glucocorticoid circulating levels in this phenotype of obesity. PMID- 15347845 TI - Estradiol and progesterone modulate the nitric oxide/cyclic gmp pathway in the hypothalamus of female rats and in GT1-1 cells. AB - Considerable evidence suggests that the nitric oxide (NO)/cGMP signaling pathway plays an important role in the expression of reproductive behavior and in gonadotropin-releasing hormone (GnRH) release from the hypothalamus The effects of the NO/cGMP pathway on GnRH release and gene expression have also been examined in GT1 cells. However, it is still controversial whether NO/cGMP signaling facilitates or inhibits GnRH release in these cells. The current study examined the effects of estradiol and progesterone on neuronal NO synthase (nNOS), soluble guanylyl cyclase (sGC), and NO-dependent cGMP production in the preoptic area (POA) and hypothalamus (HYP) as well as in GT1-1 cells. Ovariectomized female rats received vehicle, estradiol benzoate (48 h) and/or progesterone (3-4 h) before preparation of brain slices. GT1-1 cells were incubated with vehicle, estradiol (48 h), progesterone (3-4 h), or with both hormones. The combination of estradiol and progesterone increased the expression of nNOS protein in the POA and HYP. Hormones had little effect on the abundance of sGC. Estradiol and progesterone together greatly enhanced NO-stimulated sGC activity in HYP-POA slices. In GT1-1 cells, NO-stimulated sGC activity was significantly increased by estradiol and progesterone, alone or in combination, but sGC expression was not altered by hormones. PMID- 15347848 TI - ADAM 33 and its association with airway remodeling and hyperresponsiveness in asthma. AB - Asthma is known to be a Th2 inflammatory syndrome that leads to intermittent airway obstruction. However, the mechanisms involved in development of the clinical features remain enigmatic, although genetic elements clearly are involved. Recently, based on a large genome wide screen involving families in the United Kingdom and the United States with at least two siblings with asthma, a locus was identified that encoded for a family of proteases. This group of proteins is now known as the ADAM superfamily. In this review, we discuss the ADAM superfamily and, in particular, ADAM 33, a member of a family of genes which encode a subgroup of zinc dependent metalloproteinase (metzincin). The potential for therapeutic intervention with ADAM 33 is extremely attractive and further work will not only focus on the specific domains of ADAM 33, but also the mechanisms by which they lead to bronchial hyperreactivity. PMID- 15347847 TI - Airway remodeling: lessons from animal models. AB - Airway remodeling, an array of persistent tissue structural changes that occurs through a process of injury and dysregulated repair linked to airway chronic inflammation, is presently believed to largely account for the disease mechanisms of asthma. Increases in airway smooth muscle mass are probably the main mechanism causing airway hyperresponsiveness, and changes in the extracellular matrix may stimulate smooth muscle growth and contribute to the mechanics of airway obstruction. The various components of airway remodeling described inhuman asthma have been successfully reproduced in animal models of several species. Most of the data have been contributed by rat models of allergic sensitization and repeated challenge,transgenic mouse models of cytokine overexpression localized to the lung and, more recently, allergen-driven mouse models using wild-type inbred strains. Overall, animal model shave provided significant insights into the mechanisms of airway remodeling and recent technological developments allow us to exploit these models in new directions. However, the challenge of finding new therapeutic strategies that prevent or control airway remodeling,thus providing etiopathogenically oriented treatments for asthma, still stands. Experimental airway remodeling in animals should be an essential tool for treatment discovery in the near future. PMID- 15347849 TI - Differences in airway remodeling between asthma and chronic obstructive pulmonary disease. AB - The functional consequence of asthma and chronic obstructive pulmonary disease (COPD)is airflow limitation, which is mostly reversible in asthma and not fully reversible in COPD. In both diseases, inflammatory conditions are associated with cellular and structural changes,referred to as remodeling, and these structural changes may lead to thickening of the airway wall, thereby promoting airway narrowing and airflow limitation. However, the pattern of infiltrated cells and the pattern of structural changes occur differently in the two diseases. In asthma, CD4+, T lymphocytes, eosinophils, and mast cells are the predominant cells involved,whereas in COPD, CD8+, T lymphocytes, and macrophages are predominantly involved. In severe cases of asthma and COPD, neutrophil infiltration becomes evident. Regarding structural changes, epithelial injury and early thickening of reticular basement membrane are highly characteristic of the airway wall of asthmatics. Increases in airway smooth muscle mass occur in large airways of severe asthmatics and in small airways of patients with COPD. Thickening of the airway wall, goblet cell hyperplasia, mucous gland hypertrophy, and the luminal obstruction caused by inflammatory exudates and mucous are features of both asthma and COPD. Squamous epithelial metaplasia and airway wall fibrosis are commonly observed characteristics of COPD. Destruction and fibrosis of the alveolar wall occur in COPD but not in asthma. The remodeling processes accompanied by chronic inflammatory infiltrates interact in a complex fashion and contribute to the development of airflow limitation in both asthma and COPD. PMID- 15347850 TI - Clinical assessment of airway remodeling in asthma: utility of computed tomography. AB - Airway remodeling is an established feature of asthma. Histologic examination is essential in the assessment of remodeling that is a pathologic concept. Examinations of autopsied or resected lung have enabled detailed morphologic and morphometric studies and have provided fundamental knowledge of airway remodeling in asthma. However, such materials are only accidentally available, and clinical information may often be insufficient in autopsied cases. Bronchoscopic mucosal biopsy has been widely used since the 1980s, and has contributed substantially to basic investigations of inflammation and remodeling. However such specimens are limited in size and depth, limited to central airways, and the procedure might be too invasive to be repeated. Remodeling can also be assessed indirectly. Pulmonary function tests to evaluate chronic airflow obstruction are available in clinical settings and suitable for screening or mass studies, but they may be affected by concomitant diseases or short-term asthma control. Computed tomography (CT) has recently been utilized to assess remodeling. It cannot discern pathologic details but provides a broader range of airway/lung morphology and may be less invasive compared to biopsy. In addition to classic subjective evaluations,quantitative assessment has been reported for central airway dimensions, such as airway wall area, luminal area and wall thickness, and for peripheral airway abnormality or air trapping as measured by decreased lung attenuation or increased mosaic perfusion. This article summarizes the merits and limitations of various methods to assess airway remodeling, and describes the details of methodologies, interpretations, pathophysiologic relevance, and future directions of asthmatic airway remodeling assessed by CT. PMID- 15347851 TI - Impact of inhaled corticosteroids and leukotriene receptor antagonists on airway remodeling. AB - Asthma is a chronic inflammatory disease of the airways that is characterized by the presence of inflammatory cells and remodeling, a term used to define complex morphological changes involving all the structures of the bronchial wall (e.g., goblet cell hyperplasia of the epithelium, thickening of reticular basement membrane, increases of airway smooth muscle[ASM], and blood vessels). An important factor in the pathophysiology of asthma is the recognition that airway inflammation and airway remodeling are linked, as they are in other chronic inflammatory diseases. First-line therapy of persistent asthma involves the use of inhaled corticosteroids to control the underlying inflammation of the airways. Because remodeling of the airway wall is thought to be a result of chronic inflammation within the bronchial wall, it follows that because steroids reduce or reverse inflammation, they may also prevent or modulate remodeling. It has been revealed that steroids improve the subepithelial fibrosis and also significantly reduce airway vascularity. The cysteinyl leukotrienes receptor antagonists may also be helpful regarding the targeting of the inflammation and remodeling in asthma. However,long-term studies were needed to appreciate the prevention and treatment of remodeling by drug therapies. PMID- 15347852 TI - Mental health in the new millennium: research strategies for India. PMID- 15347853 TI - Open access publication in biomedical research: implications for developing countries. PMID- 15347854 TI - From smallpox to polio and beyond: disease surveillance in India. PMID- 15347855 TI - Cognitive functions in extraordinary environments. PMID- 15347856 TI - Diversity & overlap in the mechanisms of processing protein antigens for presentation to T cells. AB - The immune system needs to recognise target protein antigens from pathogens residing in both extracellular and intracellular locations. Intricate proteolytic processing events that follow antigen/ pathogen encounter provide the immune system with a complex display of a heterogeneous peptide mix, instrumental in the initiation of T cell immune responses, and allow the separation of extracellular and intracellular pathogen identification. However, recent evidence shows that this conventional dimorphism in the proteolytic processing of endogenous versus internalised antigen is less restrictive than originally recognized. The events that constitute the conventional major histocompatibility complex (MHC) restricted processing pathways are accompanied by interesting deviations that provide novel adjuncts for the processing machinery to gain access to antigen in varied intracellular locations. This review discusses these aspects of classical and non-classical processing pathways for MHC-restricted protein presentation, which play significant roles in both optimising and diversifying the peptide repertoire available for immune recognition. PMID- 15347857 TI - Communicable diseases monitored by disease surveillance in Kottayam district, Kerala state, India. AB - BACKGROUND & OBJECTIVES: A disease surveillance model developed in the North Arcot district, Tamil Nadu, was found to be practical, efficient, inexpensive and useful for public health action to monitor the success of ongoing interventions and to detect and intercept outbreaks. It was centred in the private (voluntary) sector with full co-operation and participation by the government sector. As Kerala state wanted to replicate this model in all districts, one district was chosen to pilot test it centred within the existing district public health system, soliciting participation from the private sector. A two-year (1999-2001) performance of this model is presented. METHODS: After elaborate preparations including the selection of 14 diseases to be reported and training of doctors in the private sector health care institutions and doctors and paramedical staff in all government health centres and hospitals, printed post cards were widely distributed. The business reply system was used so as to avoid handling postage stamps. Cards were received by the nodal officer in the district public health office and checked on a daily basis to detect disease prevalence and evidence of clustering in time and space. Swift action was taken on detecting case clustering. A monthly bulletin containing disease summaries and other useful information was freely distributed to all reporting centres. RESULTS: On an average, just over 100 disease reports were received every month. The most frequently reported diseases were, in the descending order, leptospirosis, acute dysentery, typhoid fever and acute hepatitis. Among vaccine-preventable childhood diseases, only measles was reported, but no diphtheria, tetanus or whooping cough. Several outbreaks were detected early and interventions applied to intercept them. The most striking example was that of cholera, the occurrence of which was detected swiftly for instituting highly successful control measures. INTERPRETATION & CONCLUSION: The district level disease surveillance system centred in the government public health system has been highly successful. Disease surveillance was responsible for the government to obtain information on the prevalence of leptospirosis in the district. The reports enabled the public health officers to detect disease-clustering as the early signals of outbreaks and to take quick remedial measures. PMID- 15347858 TI - Effect of chronic hypobaric hypoxia on components of the human event related potential. AB - BACKGROUND & OBJECTIVES: Induction to high altitude (HA) leads to deterioration in cognitive functions. The event related potentials (ERPs) like P300 are reported to be affected by hypoxia and bring about impairments in cognitive performance. The aim of the study was to investigate the effect of hypobaric hypoxia on event related potentials at two different altitudes i.e., 3200 m (HA I) and 4300 m (HA II) in ascending order to see how ERPs change with increasing altitude. METHODS: The study was carried out on 20 healthy male volunteers at sea level (SL) and thereafter at high altitude (HA) in Eastern Himalayas and on return to sea level (RSL). The P300 was recorded by using standard auditory odd ball paradigm with compact -4 (Nicolet, USA). RESULTS: The N1, N2 and P2 latencies were not significantly affected at HA I and HA II in comparison with SL indicating no effect of hypobaric hypoxia on sensory conduction. However, at HA II, most of the subjects showed an increase in latency of P3 component reflecting sensory discrimination and delay in evaluation process at 4300 m. At HA I, only 10 out of 20 subjects showed an increase in P3 wave latency and 3 did not show any change in N2-P3 components. INTERPRETATION & CONCLUSION: The results of the present study showed an increase in P300 wave latency at 3200 and 4300 m of high altitude. The observations suggest that hypoxia causes slowing of the signal processing at 4300 m, and magnitudes of the effects are altitude dependent with higher level of decline observed with increasing altitude. PMID- 15347859 TI - Correlation of mutations detected by INNO-LiPA with levels of rifampicin resistance in Mycobacterium tuberculosis. AB - BACKGROUND & OBJECTIVES: Due to emergence of drug resistance in Mycobacterium tuberculosis, there is a need to have accurate and rapid methods for detection of drug resistance to important drugs like rifampicin. The present study was aimed at evaluation of a commercially available INNO-LiPA assay, for the detection of mutation in rpoB gene region of M. tuberculosis and correlate these mutations with levels of rifampicin resistance for assessing their clinical relevance. METHODS: Fifty five well-characterized isolates of M. tuberculosis deposited from various regions of India in Mycobacterial Repository Centre at the CJILOMD, Agra were subjected to susceptibility testing for rifampicin at various concentrations of drug viz., 10, 40, 64, 128 microg/ml on Lowenstein- Jensen (LJ) medium. rpoB gene fragment (260 bp) was amplified using Rif-TB amplification kit and after hybridization, detection was done by using INNO-LiPA Rif TB kit. RESULTS: The rpoB gene could be amplified from DNA extracted from all the 55 culture isolates and showed clear hybridization pattern with M. tuberculosis complex specific probes on LiPA strips. Mutations detected were correlated with degree of rifampicin resistance. All the sensitive isolates (identified by MIC) were identified as rifampicin sensitive (100%) by INNO-LiPA as they exhibit positive for wild type 'S' probes and negative for 'R' probes. Two of the 5 isolates, resistant at 10 microg/ml and 40 microg/ml had either D516V, H526Y mutations or unknown mutations. Thirty (85.71%) isolates resistant at clinically relevant levels (64,128microg/ml) exhibited double, triple or more 'R' type mutations (R(2(D516V)), R(4a(H526Y)), R(4b(H526D)), R(5(S531L))) as well as unknown mutations present at 'S' probes region whereas remaining isolates did not show any mutation by this method. This method could identify with definitiveness 60 per cent ( 21/35) isolates as rifampicin resistant as mutations observed in others were also present in isolates with low levels of resistance. INTERPRETATION & CONCLUSION: The results indicate that INNO-LiPA Rif TB test is a rapid and easy to use method for detection of mutations associated with rifampicin resistance in M. tuberculosis. However, as some of these mutations are also present in isolates with low degree of resistance which are still microbiologically sensitive to rifampicin, there is a need to improve this assay by exclusion of some of the current probes and inclusion of more probes. PMID- 15347860 TI - Asymptomatic herpes simplex virus type 2 (HSV-2) infection among pregnant women in Turkey. AB - BACKGROUND & OBJECTIVES: A large proportion of individuals with serologic evidence of infection with herpes simplex virus type 2 (HSV-2) are asymptomatic. HSV-2 is the main cause of genital herpes infections. The acquisition of genital herpes during pregnancy has been associated with spontaneous abortion, premature labour and congenital and neonatal herpes. The present study was undertaken to determine asymtomatic genital HSV-2 shedding and seroprevalence of HSV-2 infection among asymptomatic pregnant women at the time of delivery in Adana, Turkey. METHODS: Asymptomatic 130 pregnant women without a history of genital herpes were enrolled in the study. HSV-2 shedding was determined by viral culture of the swabs collected from cervix and vulva and HSV-2 antigen was detected by direct immunofluorescence assay (IFA), HSV-2 IgG and IgM antibodies were detected by HSV-2 type specific IgG and IgM enzyme-linked immunosorbent assay (ELISA). RESULTS: HSV-2 IgG and IgM antibodies were found in 82 (63.1%) and 18 (11.3%) of 130 pregnant women. HSV-2 type-specific antigen was detected in 22 (16.9%) pregnant women by IFA test, 17 (13.1%) of whom had HSV-2 IgM antibodies. HSV-2 was isolated only in 3 women. INTERPRETATION & CONCLUSION: The seroprevalence of HSV-2 (63.1%) and genital HSV-2 infection (16.9%) was high among asymptomatic pregnant women in Adana, Turkey. Therefore, to reduce the risk of neonatal herpes, HSV-2 type-specific antibodies should be detected in pregnant women using serological tests that allow to identify women with asymptomatic or subclinical genital HSV-2 infection and those susceptible to primary genital HSV-2 infection. PMID- 15347861 TI - Current pattern in antimicrobial susceptibility of Salmonella Typhi isolates in Pondicherry. AB - Typhoid fever continues to remain a health problem as the causative organism Salmonella Typhi has developed resistance to many of the antibiotics used. This study was undertaken to determine the current pattern of resistance to antimicrobial agents and phage types of S.Typhi isolates obtained in a tertiary health care hospital in Pondicherry. Blood culture was done for 1296 suspected cases of enteric fever and 157 strains of S. Typhi were isolated. Sensitivity to ampicillin, chloramphenicol, gentamicin, ciprofloxacin and ceftriaxone was determined by disc diffusion, and the minimum inhibitory concentration (MIC) of ciprofloxacin determined. There were 61 multidrug resistant (MDR) isolates. The MIC of ciprofloxacin for 147 isolates was >0.5 mg/l; of these, 131 were resistant to nalidixic acid. Phage typing was done for 123 isolates and 115 were found to be of phage type E1, biotype 1. A decline in the number of MDR isolates was noted. Concurrently, there has been an increase in the number of isolates sensitive to all antibiotics except nalidixic acid, and all these isolates showed reduced susceptibility to ciprofloxacin. Nalidixic acid susceptibility could be a useful screening test for the detection of decreased susceptibility of S. Typhi to ciprofloxacin. The clinicians should be advised to use ceftriaxone selectively in cases showing non-responsiveness to ciprofloxacin. PMID- 15347862 TI - Effect of short-term practice of breathing exercises on autonomic functions in normal human volunteers. AB - BACKGROUND & OBJECTIVES: Practice of breathing exercises like pranayama is known to improve autonomic function by changing sympathetic or parasympathetic activity. Therefore, in the present study the effect of breathing exercises on autonomic functions was performed in young volunteers in the age group of 17-19 yr. METHODS: A total of 60 male undergraduate medical students were randomly divided into two groups: slow breathing group (that practiced slow breathing exercise) and the fast breathing group (that practiced fast breathing exercise). The breathing exercises were practiced for a period of three months. Autonomic function tests were performed before and after the practice of breathing exercises. RESULTS: The increased parasympathetic activity and decreased sympathetic activity were observed in slow breathing group, whereas no significant change in autonomic functions was observed in the fast breathing group. INTERPRETATION & CONCLUSION: The findings of the present study show that regular practice of slow breathing exercise for three months improves autonomic functions, while practice of fast breathing exercise for the same duration does not affect the autonomic functions. PMID- 15347863 TI - Time for 'publish in India' movement - a response. PMID- 15347864 TI - Permissive hypercapnia during mechanical ventilation of neonates. PMID- 15347865 TI - Effect of stepwise reduction in minute ventilation on PaCO2 in ventilated newborns. AB - OBJECTIVE: To study the effect of step reduction of expired minute ventilation (MV) on PaCO2 in ventilated newborns and to determine whether MV within a defined range can predict PaCO2. DESIGN: Prospective descriptive. SETTING: Referral neonatal unit of a teaching hospital. METHODS: Forty neonates stable on mechanical ventilation receiving minute ventilation in the range of 150-210 ml/kg/min. were studied. The spectrum of disorders for which the babies were ventilated included apnea of prematurity in 16, pneumonia in 14, meconium aspiration syndrome in 6 and hyaline membrane disease in 4. Median age at study was 6 days and median weight at study was 2.1 kgs. The MV was reduced from 210 to 150 mL/kg/min in three steps and concomitant PaCO2 was measured. Reductions were not done if PaCO2 was more than 50 mmHg. MVs were plotted against PaCO2 and a regression equation to predict PaCO2 from MV was calculated. RESULTS: A stepwise increase was seen in CO2 with reduction of MV over the range studied. The median MV and median PaCO2 achieved in the three steps were 201 mL/kg/min and 36.7 mm of Hg, 180 mL/kg/min and 41.7 mm of Hg, 160 mL/kg/min, and 44.3 mm of Hg. The regression equation to predict PaCO2 was PaCO2 = 70 - 0.17 x MV in mL/kg/min, r = -0.45, r2 = 0.20, residual variance (s2) = 39.37; gave a predicted PaCO2 within 12.5 mmHg. for a given MV. CONCLUSION: Reducing minute ventilation led to an increase in the levels of PaCO2. Minute volumes of 160 ml/kg/min correlated with PaCO2 value of 44.3 mm of Hg. MV as low as 160 mL/kg/min are well tolerated by newborns. PMID- 15347866 TI - Management of edema in nephrotic syndrome. AB - Nephrotic syndrome is a recurrent or chronic disorder characterized by heavy proteinuria, hypoalbuminemia and edema. In most patients, the edema can be managed satisfactorily with judicious oral administration of loop diuretics, most often frusemide. The treatment of patients with intractable edema is more difficult and comprises a series of measures including salt restriction, treatment with intravenous frusemide, additional use of thiazide and potassium sparing diuretics, and intravenous albumin. Ultrafiltration and head out water immersion are beneficial for treatment of patients with edema, which is unresponsive to the above measures. PMID- 15347867 TI - Human platelet specific antigens and their importance. PMID- 15347868 TI - Iron formulations in pediatric practice. PMID- 15347869 TI - Anterior lens capsule vascularity in evaluating gestation in small for gestation neonates. AB - This study evaluated the role of anterior lens capsule vascularity in assessing gestation in small for gestational age (SGA) neonates in a prospective manner. Neonates with birth weight less than 2000 g were recruited. Those with gross congenital malformations, including corneal opacity or cataract and evidence of TORCH infection were excluded from the study. A total of 139 subjects were enrolled into the study (60-appropriate for gestational age (AGA) and 79 small for gestational age (SGA) neonates). Clinical gestational age assessment and ophthalmoscopic examination for anterior lens capsule vascularity grading were done within 24 hours of birth by independent blinded observers. The correlation of lens capsule grading with clinical gestation in AGA neonates (r2 = 0.75) was stronger compared to SGA neonates (r2 = 0.50). Gestational age assessment by lens capsule grading had a better agreement with clinical gestation in AGA subjects (Kappa Index - 0.56 in AGA vs. 0.26 in SGA). The lens capsule grading under estimated the gestation of SGA subjects by an average of three weeks compared to AGA neonates. PMID- 15347870 TI - Spectrum of atypical celiac disease in North Indian children. AB - Atypical celiac disease (ACD) presenting in childhood has rarely been documented from India. The present retrospective study analyzed features of atypical celiac disease over a 5-year period. Patients were diagnosed to have Celiac Disease (CD) as per the standard ESPGHAN criteria. The biochemical and hematological parameters of the cohort of children presenting with atypical features (ACD) were compared with children presenting as typical diarrheal CD. Twelve children were diagnosed to have CD. Seven of them presented with ACD. The two groups did not differ significantly in their age of presentation, hematological and biochemical profile. Osteoporosis as documented on bone mineral densitometry was present in all 6 patients of ACD in whom BMD was done. Short stature (4) and refractory iron deficiency anaemia (3) was the commonest modes of presentation of ACD. Occurrence of these conditions either singly or in combination warrants exclusion of celiac disease in children. PMID- 15347871 TI - Serum prolactin in seizure disorders. AB - This study aimed to determine the post-ictal prolactin (PL) response in different types of seizures and seizure-like events in children, and correlate with the post-ictal duration. Patients were divided into group I (generalized tonic-clonic seizures, complex partial seizures or simple partial seizures), group II (febrile convulsions) and group III (conditions mimicking seizures). Group IV consisted of 25 controls. Blood was collected within 2 hours of the seizure and PL levels assayed. PL levels were significantly high only within group I; highest and baseline levels were attained within 10 minutes and by 100 minutes respectively. The sensitivity and specificity of elevated PL for epileptic seizures were 64 percent and 98 percent respectively. It is concluded that a high prolactin level within 100 minutes of a seizure is suggestive that a generalized or complex partial seizure has occurred. PMID- 15347872 TI - Spectrum of congenital CNS malformations in pediatric epilepsy. AB - This study was conducted in a tertiary pediatric epilepsy clinic to ascertain the spectrum of development malformations in children, with seizures. Seventy Six Children (0-12 yr) with seizures and CNS malformations based on neuroimaging were included. Observed anomalies included dysgenetic corpus callosum (DCC), lissencephaly, focal cortical dysplasia (FCD), pachygyria, polymicrogyria, heterotopia, schizencephaly, holoprosencephaly, hemimegalencephaly, and phakomatoses like tuberous sclerosis, Sturge Weber syndrome and linear cutaneous nevus syndrome. Seizure semiology varied in all categories. Microcephaly, developmental delay and tone abnormalities were common clinical findings. 60.5 percent cases presented in infancy. The characteristic EEG features provided a clue to the diagnosis of anomalies like lissencephaly, agenesis of corpus callosum and alobar holoprosencephaly. PMID- 15347873 TI - Fetal rhabdomyoma in a neonate. AB - Rhabdomyoma is a rare benign tumour, majority arising from the cardiac muscle. Seventy to 90 percent of extra cardiac rhabdomyomas are found in the head and neck region, usually within the upper aero digestive tract. We report a case of rhabdomyoma of anterior neck in a neonate. Although rhabdomyomas of posterior neck have been reported, those reported in anterior triangle are infrequent. The lesion has not recurred one year after complete excision. There are no similar reports in Indian literature. PMID- 15347874 TI - Hyperammonemia with citrullinemia. AB - Two cases of hyperammonemia with elevated citrulline are reported, one resulting from a deficiency of pyruvate carboxylase and the other from a partial deficiency of argininosuccinate synthetase. Diagnosis was based on clinical, biochemical and amino acid profiles. The utility of amino acid determinations in hyperammonemia suspected to underlie an inborn error of metabolism is emphasized. PMID- 15347875 TI - VACTERL association with Prune-Belly syndrome. AB - We report a term, small for gestational age neonate having full spectrum of VACTERL association. In addition, the neonate also had triad of signs and symptoms associated with prune belly syndrome. The concurrence of these two syndromes could lie in their common etiology of defect in mesodermal differentiation. Such a combination is extremely rare and is generally incompatible with life. PMID- 15347876 TI - Caroli's disease. AB - Carolis disease is a rare communicating segmental or diffuse dilatation of the intrahepatic biliary tree. Cholangitis, liver cirrhosis and cholangiocarcinoma are its potential complications. A case of Carolis disease in a boy of 6 years with bilobal involvement presenting with intermittent abdominal pain, fever and hepatomegaly is reported here. PMID- 15347877 TI - Congenital lumbar hernia. PMID- 15347878 TI - Yellow skin papules over the neck-pseudoxanthoma elasticum. PMID- 15347879 TI - Prevalence of obesity in affluent school boys in Pune. PMID- 15347880 TI - Embryonal rhabdomyosarcoma of lower lip. PMID- 15347881 TI - Monitoring adverse drug events: need for an active surveillance system. PMID- 15347882 TI - Role of Entamoeba histolytica in acute watery diarrhea in hospitalized under-five children. PMID- 15347883 TI - Giant occipital meningocele as a presenting feature of Dandy-Walker syndrome. PMID- 15347884 TI - Cost of syrup versus capsule form of vitamin A. PMID- 15347886 TI - Ellis-Van Crevald syndrome. PMID- 15347887 TI - Utility of Interphase FISH Panels for Routine Clinical Cytogenetic Evaluation of Chronic Lymphocytic Leukemia and Multiple Myeloma. AB - Specific genetic abnormalities are of prognostic significance for patients with chronic lymphocytic leukemia (CLL) and multiple myeloma (MM); however, routine cytogenetic analysis usually provides normal results. We utilized two probe panels for interphase fluorescence in situ hybridization (FISH) studies to enhance the ability to detect genetic abnormalities in samples that were referred for routine cytogenetic studies for possible diagnoses of CLL or MM. The CLL panel consisted of probes for 11q22.3 (ATM gene), 13q14 (D13S319), the centromere of chromosome 12 (D12Z3) and 17p13.1 (P53 gene). The MM panel included probes for 14q32 (IgH gene) and/or t(11:14)(q13;q32) (BCL1/IgH), 13q14 (D13S319) and 17p13.1 (P53 gene). FISH detected clonal aberrations not identified by conventional cytogenetics in an additional 8 of 23 (35%) samples referred for possible CLL and 7 of 42 (17%) samples with possible MM. The prognostic significance of the aberrations identified ranged from favorable, to intermediate, to poor. Our studies indicate that many samples referred for routine cytogenetics testing for CLL and MM yield normal results for both conventional and FISH testing, likely due to lack of definitive diagnosis in a percentage of cases. However, FISH is more sensitive for the detection of clinically significant chromosome abnormalities and should be the testing methodology of choice for these disorders. PMID- 15347888 TI - Prognostic impact of EBV-related LMP-1, histologic type, and environmental factors in nasopharyngeal carcinoma in a German population. AB - INTRODUCTION: This retrospective study addressed the possible involvement of latent Epstein-Barr virus (EBV) infection, in particular LMP-1 expression, and further exogenous factors, i.e. tobacco, alcohol and occupational hazardous substances, in nasopharyngeal carcinoma (NPC) in a German population. PATIENTS AND METHODS: From 1980 to 2000, 44 patients suffering from histologically confirmed NPC were entered into the study. 33 specimens were available for immunostaining (IHC) to analyze LMP-1 expression. Information about environmental exposures were obtained employing a detailed standardized questionnaire. RESULTS: Outcome of patients with squamous cell NPC (SC-NPC) was significant worse than that of those with non-keratinizing NPC (NK-NPC). Age and tumor size correlated with response to therapy. The group with negative conventional LMP-1 staining showed better overall survival after 5 years compared to the group with positive or marginally positive LMP-1 detection (not significant). Nevertheless, after staining by tyramid-augmented IHC (TSA-IHC), nearly all specimens with negative LMP-1-staining in conventional IHC were found to be clearly positive. All patients with SC-NPC were smokers. The distribution of smokers and non-smokers in the group of NK-NPC was balanced. Comparable to the tobacco observation, there was also a correlation between high alcohol consumption and SC-NPC. CONCLUSION: Prognosis of NPC is mainly dependent on histologic type. Prognostic impact of LMP 1 is still unclear since LMP-1 was detected in all specimens using TSA-IHC. Therefore, TSA-IHC-LMP-1 detection might be interesting for diagnostic specification and development of new therapeutic strategies in NPC. PMID- 15347889 TI - Differential regulation of MAPK (JNK 3) gene expression in human head and neck squamous cell carcinomas. AB - BACKGROUND: The JNK 3 gene encodes a protein which belongs to the mitogen activated protein (MAP) kinases, a group of molecules involved in signaling pathways. MATERIAL AND METHODS: Messenger RNA extracted from head and neck squamous cell carcinoma (HNSCC) cells and normal upper aerodigestive tract mucosa keratinocytes was reversely transcribed. The resulting cDNA populations were subjected to an arbitrarily primed mRNA fingerprint. After electrophoresis, the band pattern was detected by autoradiography. RESULTS: A 107 bp mRNA fragment was detected in HNSCC cells showing considerable repression in comparison with the benign phenotype. After cloning of this fragment, a database search revealed an exact homology with sequences belonging to the c-jun N-terminal MAP-10 kinase (JNK 3). Northern hybridization confirmed the distinctly reduced expression of this gene in HNSCC biopsies in contrast to adjacent normal mucosa. CONCLUSION: The results show evidence that the expression of the JNK 3 gene is strongly repressed, suggesting that JNK 3 is implicated in carcinogenic processes in head and neck cancer. PMID- 15347890 TI - Clinical relevance of circulating tumour cells in the bone marrow of patients with SCCHN. AB - BACKGROUND: Clinical outcome of patients with head and neck squamous cell carcinoma (SCCHN) depends on several risk factors like the presence of locoregional lymph node or distant metastases, stage, localisation and histologic differentiation of the tumour. Circulating tumour cells in the bone marrow indicate a poor prognosis for patients with various kinds of malignoma. The present study examines the clinical relevance of occult tumour cells in patients suffering from SCCHN. PATIENTS AND METHODS: Bone marrow aspirates of 176 patients suffering from SCCHN were obtained prior to surgery and stained for the presence of disseminated tumour cells. Antibodies for cytokeratin 19 were used for immunohistochemical detection with APAAP on cytospin slides. Within a clinical follow-up protocol over a period of 60 months, the prognostic relevance of several clinicopathological parameters and occult tumour cells was evaluated. RESULTS: Single CK19-expressing tumour cells could be detected in the bone marrow of 30.7% of the patients. There is a significant correlation between occult tumour cells in the bone marrow and relapse. Uni- and multivariate analysis of all clinical data showed the metastases in the locoregional lymph system and detection of disseminated tumour cells in the bone marrow to be statistically highly significant for clinical prognosis. CONCLUSION: The detection of minimal residual disease underlines the understanding of SCCHN as a systemic disease. Further examination of such cells will lead to a better understanding of the tumour biology, as well as to improvement of diagnostic and therapeutic strategies. PMID- 15347891 TI - Management of N0 neck in head and neck cancer: current controversies. AB - Squamous cell carcinomas of the upper aerodigestive tract are characterized by a high frequency of lymphogenic metastatic spread. The neck should be included in the treatment concept because almost 25% of all patients develop occult lymph node metastases. The only exception are early carcinomas of the lower lip and the vocal cords. Since currently for high risk patients no prognostic markers or other factors for occult metastatic spread are available, treatment strategies have to include the exact knowledge of direction and extent of regional lymphatic drainage. Based on this prerequisite, concepts for selective cervical lymph node dissection in case of the clinical N0 situation were developed which may also be used for an individually optimized radiotherapy. Intensity modulated radiotherapy (IMRT) bears a high potential for an optimized irradiation in case of potential occult metastatic spread of carcinomas of the upper aerodigestive tract. Without compromising local control rates IMRT allows a considerable reduction of permanent xerostomia by sparing at least one major salivary gland. The significant scientific purpose for the N0 neck, however, will remain the identification of high-risk patients, e.g. by means of tissue and/or serum related markers, which carry a high risk of local and distant metastases. PMID- 15347892 TI - Approaches to preserve larynx function in locally advanced laryngeal and hypopharyngeal cancer. AB - Randomized controlled studies have shown that preservation of the larynx function in patients with advanced resectable laryngeal and hypopharyngeal cancer is possible without compromising survival compared to total laryngectomy (TL). Options for preserving the larynx include function-sparing surgery, radiotherapy alone, induction chemotherapy followed by radiotherapy of responders, and concomitant radiochemotherapy. The current data suggest that induction chemotherapy followed by radiotherapy of responders is an acceptable alternative to TL for patients desiring larynx preservation. Concomitant radiochemotherapy (platinum/5-FU) leads to superior local control and larynx preservation rates compared to induction chemotherapy followed by radiation. The optimal treatment sequence for newer cytotoxic agents is, however, unclear. Such cytotoxic agents and more effective fractionation regimens as well as more advanced surgical techniques are currently evaluated. Predictive tests to successfully stratify patients for the optimal treatment option and more effective systemic therapy are needed to improve therapeutic possibilities and survival. PMID- 15347893 TI - Lung cancer incidence in the Czech Republic: a time-trend study. AB - BACKGROUND: Analysis of cancer incidence trends is one of the prerequisites for cancer prevention. It is possible to set up priorities and assess current development of morbidity. METHODS: All lung cancer incidences in the Moravia region of the Czech Republic (4,000,000 inhabitants) in the years 1984-1998 have been assessed concerning incidence, anatomical subsites, stage, and morphology. Age and sex distribution have been taken into consideration. RESULTS: There has been a continuous increase in the incidence of lung cancer in females and a mild decrease in males. In both sexes the most affected anatomical subsites were upper and lower lobe of the lung. Stage III and IV predominated. Main morphological type was squamous cell carcinoma in males (39.3%) and in females (22.2%), the second most frequent type in males was small cell carcinoma (13.6%) and in females adenocarcinoma (21.8%). The highest incidence was observed in the age group of 55-75 in males and 60-80 in females. CONCLUSION: Lung cancer epidemiology and prevention is one of the top-priority health problems in the Czech Republic. The incidence is one of the highest in the world especially for males; in females it is continuously increasing. Primary prevention is available but difficult to implement, particularly non-smoking. Secondary prevention is not available due to lack of suitable screening tests and a short lead time. PMID- 15347894 TI - Outpatient therapy in metastatic breast cancer. A retrospective study of 90 patients treated in an oncology group practice. AB - BACKGROUND: Whenever possible, treatment of breast cancer should be performed in an outpatient setting, but only few data about patients being treated exclusively on an outpatient basis are available. PATIENTS AND METHODS: A retrospective analysis was performed in 90 unselected patients who were treated consecutively in our oncology group practice between 6/95 and 8/99. RESULTS: Median age at detection of metastases was 55 years (30-90) and performance status ranged from 0 to 2. 83 patients (92.2%) received chemotherapy, 7 (7.8%) received endocrine therapy only. CMF was used in 27.7%, anthracyclines in 71.1% and taxanes in 43.3%. 855 chemotherapy cycles were performed and the observed toxicity was mild. Reversible grade 3 and 4 hematotoxicity was seen after 27 cycles (3.2%). Neurotoxicity or mucositis grade 3 and 4 were seen in 6 patients (6.6%). Therapy associated hospitalization occurred in 1 patient thrice due to febrile neutropenia. Complete remissions were seen in 5 patients (5.6%) during first-line therapy. Median survival of the whole cohort until the end of the follow-up period (2/03) was 28 months (2-259). Overall survival after 1, 2, 3 and 5 years was 83, 56, 33 and 18% respectively. 50% could die at home. CONCLUSIONS: Treatment of metastatic breast cancer can be performed with minimal toxicity and a high patient acceptance in the outpatient setting. Overall survival and median survival are comparable to historical results achieved in specialized academic hospitals. Hospitalization could be avoided in the majority of patients and half of them could die at home. PMID- 15347895 TI - Weekly paclitaxel combined with local hyperthermia in the therapy of breast cancer locally recurrent after mastectomy--a pilot experience. AB - BACKGROUND: The combination of chemotherapy and hyperthermia (HT) is a promising approach in the treatment of malignant tumors. In the present report we evaluate the efficacy and toxicity of a combination of weekly paclitaxel combined with local hyperthermia in breast cancer. PATIENTS AND METHODS: 7 patients were treated for inoperable local recurrence of breast cancer after mastectomy, irradiation, and chemotherapy or hormonal therapy. They weekly received paclitaxel (60-80 mg/m(2)) in 3-h infusions followed by local HT 41-44 degrees C for 45 min for 6-18 cycles. RESULTS: Objective local response was observed in all treated patients (complete response in 4 patients and partial response in 3 patients). There were no grade 3 or 4 toxicities, neurologic toxicity or hypersensitivity reactions. Local tolerance to this regimen was also good, with only 4 patients developing mild transient erythema. CONCLUSION: Our experience indicates that the combination of weekly paclitaxel and HT may be effective in the treatment of locally recurrent breast cancer after mastectomy. PMID- 15347896 TI - Toxic epidermal necrolysis after cranial radiotherapy and phenytoin treatment. AB - BACKGROUND: There are few reports about Stevens-Johnson syndrome, a bullous form of erythema multiforme, that can develop in patients treated with cranial irradiation and antiepileptic drugs, especially with phenytoin. We present a patient who developed toxic epidermal necrolysis, a rare and severe form of Stevens-Johnson syndrome, during cranial radiotherapy and phenytoin treatment. CASE REPORT: A 65-year-old male patient with stage IIIB non-small cell lung carcinoma developed a brain metastasis. The patient was treated with phenytoin and dexamethasone. Palliative total cranial irradiation was performed. On the 23rd day of phenytoin administration, erythema and edema in the radiotherapy area and lips, as well as widespread maculopapular eruptions and rashes in the upper thoracic area were observed. The dermal lesions progressed to bullae and subsequently toxic epidermal necrolysis covering 70% of the whole body surface developed. The patient died within 15 days of appearance of the lesions due to secondary infections, despite supportive and symptomatic treatment. CONCLUSION: Although toxic epidermal necrolysis is a rare toxicity it must always be considered during cranial irradiation and antiepileptic prophylaxis. PMID- 15347897 TI - Aggressive management of recurrent ovarian cancer--the challenge of individualizing cancer therapy illustrated by a case report. AB - BACKGROUND: In clinical practice, treatment recommendations and the patient's wishes often diverge, facing the physician with difficult choices. CASE REPORT: The clinical course of a 36-year-old patient with 'platinum-refractory' ovarian cancer is reported. The patient experienced a symptomatic relapse 7 months after debulking surgery and completion of platinum-based first-line chemotherapy. As she had given birth to a son 22 months before diagnosis, she fought with outmost determination against her disease. Her husband supported her, and both asked for maximal therapy, including intensive care treatment for recurrent respiratory tract infections and total parenteral nutrition (TPN). For the patient, it was of major importance to stay with her family and make sure that her son would be able to remember his mother. Problems related to TPN and progression of disease affected her individual perception of quality of life to a much lower extent than expected and perceived by her caretakers. All professional health care providers were more than once very reluctant to continue treatment and only after extensive counseling gave in to the demand of the patient for further treatment, considering the effort futile - only to be surprised by treatment response and recovery. After 3 years of palliation, the tumor was resistant to all cytotoxic regimens and the patient died 2 months after withdrawal of chemotherapy. CONCLUSION: This case report illustrates that also in the age of evidence-based medicine individualized treatment beyond proven strategies can offer patient benefit. Taking the child's development into account makes it impossible to determine the cost-benefit ratio. PMID- 15347898 TI - Gallbladder metastasis of non-small cell lung cancer. AB - BACKGROUND: Although non-small cell lung cancer is known for its potency to spread to almost any organ of the body, metastasis to the gallbladder with significant clinical manifestation is rarely reported in the literature. CASE REPORT: We report the case of a 45-year-old man with non-small cell lung cancer (NSCLC), who developed symptoms of acute cholecystitis caused by a metastasis of the gallbladder wall. Histological examination showed tumor cell invasion in regional gallbladder lymph nodes. A second primary tumor of the gallbladder was excluded by immunohistochemical methods. CONCLUSION: Our experience showed that acute cholecystitis can occur in association with metastases of lung cancer to the gallbladder. PMID- 15347899 TI - The spectrum of mesenchymal skin neoplasms reflected by the new WHO classification. AB - Mesenchymal tumors are a heterogeneous group of tumors often arising in the skin and soft tissue. The tumors have been reclassified by the WHO in 2002. Benign mesenchymal tumors are about a hundred times more frequent than malignant mesenchymal tumors. Clinically, mesenchymal tumors often present as skin-colored nodes. Overall, elderly persons are more affected than younger individuals. The etiology is often unknown, sometimes there is an association with insults such as radiation, scars, or lymphedematous or venous stasis. Whereas some years ago a wide excision with a margin of 3-5 cm was performed for malignant variants, today micrographic surgery is preferred, as it can avoid mutilating procedures. Early detection and removal is critical since mesenchymal skin tumors often cannot be cured by radiation or chemotherapy. PMID- 15347900 TI - Photodynamic therapy--a new treatment option for epithelial malignancies of the skin. AB - Photodynamic therapy (PDT) is based on the activation of a photosensitizer by illumination with visible light, leading to photochemical tissue destruction or immunomodulation. The greatest disadvantage of systemic administration of photosensitizers is cutaneous photosensitization of the patients, which lasts for some months. An alternative approach for epithelial precancerous lesions, such as actinic keratoses and Bowen's disease, as well as for superficial skin tumors, such as basal cell carcinomas, is the topical application of sensitizers, especially 5-aminolevulinic acid. Topical PDT causes selective tissue necrosis and tumor destruction and produces excellent cosmetic results. The following article summarizes the main principles of PDT and provides a synopsis of the present status of the use of PDT for epithelial skin malignancies. PMID- 15347902 TI - [Circular instead of hierarchic]. PMID- 15347901 TI - High-volume centers--effect of case load on outcome in cancer surgery. AB - The detailed analysis of the literature of the past 10 years comprising 12 papers concerning pancreatic cancer and 10 dealing with esophageal cancer show a clear reduction of postoperative mortality with increasing case volume per year. Single papers have analyzed the main reasons for this phenomenon and showed that postoperative complication rates are lower in high-volume hospitals and their management of complications is more successful. Further long-term prognosis is also correlated to case volume. Surgery of pancreatic and esophageal cancer is a task for high-volume hospitals because they have less postoperative mortality and improved long-term prognosis compared to low-volume hospitals. PMID- 15347903 TI - Coping strategies and the request for a consultation on complementary and alternative medicine -- a cross-sectional survey of patients in a psychosomatic and three medical departments of a german university hospital. AB - OBJECTIVE: To identify factors, predicting which patients of a large German university hospital request for a consultation on complementary and alternative medicine (CAM). PATIENTS AND METHODS: 350 patients of 3 medical and a psychosomatic department of the University Hospital Freiburg were consecutively surveyed with respect to their coping strategies, health locus of control, anxiety, depression, and the request for a consultation on CAM. Predicting factors were identified via logistic regression models. RESULTS: The request for a CAM consultation was primarily determined by an active coping strategy (odds ratio 1.66, 95% confidence interval 1.27-2.17, p = 0.0002) and a low Karnofsky index (odds ratio 0.79, 95% confidence interval 0.62-0.99, p = 0.0417). Anxiety, depression or internal health locus of control correlated with each other but could not be proved as independent factors for the request for a CAM consultation. CONCLUSION: Patients of a university hospital with an active disease coping have a need for information about CAM. PMID- 15347904 TI - Patient-Practitioner-Remedy (PPR) Entanglement, Part 7: a gyroscopic metaphor for the vital force and its use to illustrate some of the empirical laws of homeopathy. AB - BACKGROUND: One of the principle obstacles to homeopathy's general acceptance has been its perceived lack of sound theoretical basis within accepted deterministic bio-medical thought. This impasse might be circumvented if instead, appeal was made to the nondeterministic concepts of the physical sciences, e.g., quantum theory and its notions of entanglement, nonlocality, and uncertainty; Weak Quantum Theory (WQT) and Patient-Practitioner-Remedy (PPR) Entanglement representing two new complementary strands of thought with the potential to create a new theoretical basis for homeopathy. OBJECTIVE: The goal of this present study was to generate a preliminary mathematical model of the action and reaction of the Vital Force to diseases and remedies within the developing contexts of WQT and PPR Entanglement, based on the metaphor of a hypothetical 'quantized' gyroscope as its physical representation. METHODS: The physics of gyroscopic motion was combined with the quantum theory describing rotating objects (without some of its imposed limitations, e.g., Planck's constant, in line with the relaxation of some of orthodox quantum theory's axioms as proposed by WQT). Thus, increase or decrease in the rate of spin of the Vital Force's hypothetical gyroscope was described in terms of quantized 'shift operators' constructed mathematically from the known 'complementarity' of a remedy's primary and secondary symptoms, expressed in the notation of complex numbers. Ultimately, this generates a hypothetical 'wave function' for the Vital Force. RESULTS: This hypothetical 'wave function' has been used to illustrate certain empirical observations of homeopathy and conventional medicine, e.g., the biphasal action of remedies encapsulated in the Arndt-Schulz Law, Wilder's Law of Initial Value, and some of the results of homeopathic provings. CONCLUSION: This preliminary theoretical analysis suggests that perhaps these less well-known empirical observations should be reinvestigated and, if confirmed, could begin ultimately to provide a much-needed alternative to the doubleblind placebo-controlled trial as a means of investigating and testing the efficacy of homeopathy. PMID- 15347905 TI - Research into complementary and alternative medicine across Europe and the United States. AB - BACKGROUND: There has been an increase in complementary/ alternative medicine (CAM) research with a wide range of perspectives. OBJECTIVE: To capture and analyse the diversity of CAM research across 7 European countries (Germany, United Kingdom, Italy, France, Spain, Netherlands, Belgium) and the US. We were predominantly interested in finding out how CAM research differs between countries. STUDY DESIGN: A systematic review of CAM literature published in 2002 and included in Medline, Embase, International Bibliography of the Social Sciences (IBBS) and PsycLit. OUTCOME CRITERIA: Data were extracted regarding country of origin, type of methodology, type of research question, CAM modality, direction of conclusion, setting and type of journal. RESULTS: In total 652 abstracts of articles were assessed (Germany: 137, UK: 183, Italy: 39, France: 47, Spain: 24, Netherlands: 17, Belgium: 22, US: 183). The vast majority of CAM research was of a medical nature and published in medical journals. The majority of articles were nonsystematic reviews and comments, analytical studies and surveys. The UK carried out more surveys than any of the other countries and also published the largest number of systematic reviews. Germany, the UK and the US covered the widest range of interests across various CAM modalities and investigated the safety of CAM. CONCLUSION: Our data suggest that important national differences exist in terms of the nature of CAM research. This raises important questions regarding the reasons for such differences. PMID- 15347906 TI - [Naive empiricism?]. PMID- 15347907 TI - Surveillance for hospitalizations with infection-related diagnoses after chemotherapy among breast cancer patients diagnosed before age 65. AB - BACKGROUND: The aim of this study was to assess the feasibility of population based surveillance of the frequency of (and risk factors for) hospitalization with infection-related conditions potentially attributable to chemotherapy among breast cancer patients diagnosed before age 65 years in the USA, where a previous report was limited to older patients. METHODS: This pilot study involved a sample of 675 Connecticut women diagnosed before the age of 65 years with early-stage breast cancer in 1999, who were identified from a population-based statewide cancer registry and were linked with a statewide inpatient hospital discharge database. RESULTS: Hospitalization with an infection-related condition (neutropenia, fever, and/or infection/bacteremia) after diagnosis was found for 8.6% of 463 patients with chemotherapy vs. 2.8% of 212 without chemotherapy (p < 0.01). Such hospitalization was positively associated with lymph-node status and receipt of chemotherapy. CONCLUSION: Population-based surveillance appears feasible, but larger US studies are needed, with additional information (including use of antibiotics) as outlined by the European Surveillance of Infections in Cancer Patients (ESIC) study of risks and risk factors for serious adverse outcomes after cancer treatment. PMID- 15347908 TI - Progression of cisplatin-induced nephrotoxicity in a carnitine-depleted rat model. AB - BACKGROUND: This study has been initiated to investigate whether endogenous carnitine depletion and/or carnitine deficiency is an additional risk factor and/or a mechanism in cisplatin-induced nephrotoxicity and to gain insights into the possibility of a mechanism-based protection by L-carnitine against this toxicity. METHODS: 60 male Sprague-Dawley rats were divided into six groups of 10 animals each and received one of the following treatments: The first three groups were injected intraperitoneally with normal saline, L-carnitine (500 mg/kg), and D-carnitine (750 mg/kg), respectively, for 10 successive days. The 4th, 5th, and 6th groups were injected intraperitoneally with the same doses of normal saline, L-carnitine and D-carnitine, respectively, for 5 successive days before and after a single dose of cisplatin (7 mg/kg). Six days after cisplatin treatment, the animals were sacrificed, and serum as well as kidneys were isolated and analyzed. RESULTS: A single dose of cisplatin resulted in a significant increase in blood urea nitrogen (BUN), serum creatinine, malondialdehyde (MDA) and nitric oxide (NO) and a significant decrease in total carnitine, reduced glutathione (GSH) and adenosine triphosphate (ATP) content in kidney tissues. Interestingly, L carnitine supplementation attenuated cisplatin-induced nephrotoxicity manifested by normalizing the increase of serum creatinine, BUN, MDA and NO and the decrease in total carnitine, GSH and ATP content in kidney tissues. In the carnitine depleted rat model, cisplatin induced a progressive increase in serum creatinine and BUN as well as a progressive reduction in total carnitine and ATP content in kidney tissue. Histopathological examination of kidney tissues confirmed the biochemical data, i.e. L-carnitine supplementation protected against cisplatin induced kidney damage, whereas D-carnitine aggravated cisplatin-induced renal injury. CONCLUSION: Data from this study suggest that: (1) oxidative stress plays an important role in cisplatin-induced kidney damage; (2) carnitine deficiency should be viewed as an additional risk factor and/or a mechanism in cisplatin induced renal dysfunction, and (3) L-carnitine supplementation attenuates cisplatin-induced renal dysfunction. PMID- 15347909 TI - Cytogenetic consequences of vinblastine treatment in mouse bone marrow. AB - BACKGROUND: Vinblastine (VBL), a vinca alkaloid, has very often been included in different cancer chemotherapeutic treatment regimens. Chemotherapy cures certain cancers and, at least, increases the life expectancy of cancer patients. However, in cancer survivors, a second malignancy frequently occurs after chemotherapy, which warrants detailed genotoxicity testing of the chemotherapeutic agents. The available genotoxicity test reports on VBL are self-contradictory and inconclusive. Thus, following a suitable experimental protocol, it is necessary to test the cytogenetic consequences of VBL treatment in mammals. METHODS: Swiss mice received 1 of 3 different doses of VBL (0.5, 1.0 and 1.5 mg/kg body weight) as a single intraperitoneal injection. The cytogenetic toxicity of VBL was assessed from the induced aberrant metaphases, chromosomal aberrations (CAs) excluding gaps and the mitotic index (MI) 24 h after treatment, and micronuclei (MN) 30 h after treatment. RESULTS: All 3 doses of VBL induced statistically significant (p < or = 0.01) percentages of aberrant metaphases and CAs, but there was no significant change in the MI. The induced percentage of aberrant metaphases and CAs were decreased with the increase in the dose of VBL. On the other hand, there was a dose-dependent and significant (p < or = 0.01) increase in MN induction. CONCLUSIONS: The results of this study indicate the clastogenic potential of VBL in the mouse bone marrow. In the present study, the induction of numerous relatively large-sized MN by VBL is in agreement with the reported aneugenic action of the drug. Although VBL is cytotoxic and is a spindle poison, the mechanism(s) involved in bringing about its clastogenic effects is yet to be elucidated. PMID- 15347910 TI - Conditioning regimens including high-dose busulfan cause a high incidence of transplant-related mortality after myeloablative stem cell transplantation. AB - BACKGROUND: Transplant-related mortality (TRM) significantly decreases the long term survival of patients with hematopoietic diseases who undergo hematopoietic stem cell transplantation (HSCT). METHODS: We retrospectively evaluated the risk factors for TRM of 58 consecutive patients with hematopoietic disease who underwent allogeneic myeloablative HSCT between April 1994 and June 2002. RESULTS: Thirty-one patients died after HSCT, 16 of whom were diagnosed as TRM. The actuarial incidence of TRM was 57.6 % in patients who received conditioning chemotherapies including high-dose busulfan and 19.4 % in those who did not (p = 0.008). Multivariate Cox model analysis revealed that disease status, graft versus host disease (GVHD) grades III-IV and the use of high-dose busulfan were significant and independent risk factors for TRM. CONCLUSION: These results suggest that high-dose busulfan as a conditioning therapy for allogeneic HSCT should be used cautiously, especially in patients with advanced disease. PMID- 15347911 TI - Antimicrobial resistance of Streptococcus pneumoniae isolates of outpatients in Germany, 1999-2000. AB - BACKGROUND: The aim of the study was to determine the prevalence of antimicrobial resistance among clinical isolates of Streptococcus pneumoniae during the winter of 1999-2000 in Germany. METHODS: Pneumococcal isolates were prospectively collected by 14 different clinical microbiology laboratories. Minimal inhibitory concentrations of penicillin G, erythromycin A, clarithromycin, roxithromycin, azithromycin, clindamycin, levofloxacin and telithromycin were determined by the broth microdilution method. RESULTS: Among 328 strains 4.6% were nonsusceptible to penicillin G (intermediate and resistant strains) and 9.5% were resistant to erythromycin A. Analysis of erythromycin-resistant strains for the underlying resistance determinants revealed that 12 (38.7%) belonged to the erm(B) and 19 (61.3%) to the mef(E) type of resistance. Among the macrolide-resistant strains, serotypes 19F (n = 9) and 14 (n = 8) were the predominant types. CONCLUSIONS: Macrolide resistance in Germany is of growing concern and mainly due to the high prevalence of pneumococci expressing the mef(E) type of resistance. PMID- 15347912 TI - Effects of dexamethasone, all-trans retinoic acid, vitamin D(3) and interferon alpha on FO myeloma cells. AB - BACKGROUND: Since multiple myeloma responds poorly to conventional chemotherapy or radiotherapy, new therapeutic approaches are needed. This study investigated the effects of dexamethasone, all-trans retinoic acid (ATRA), the active metabolite of vitamin D(3) [1,25(OH)(2)D(3)] and interferon-alpha on FO mouse myeloma cells (non-immunoglobulin-secreting myeloma cell line) in single drug or drug combination groups in vitro. METHODS: Apoptosis ratio and change in cell counts in 4 single drug groups (dexamethasone, ATRA, vitamin D(3) and interferon alpha) and 6 combination drug groups (dexamethasone + vitamin D(3,) dexamethasone + ATRA, dexamethasone + interferon-alpha, vitamin D(3) + ATRA, vitamin D(3) + interferon-alpha, interferon-alpha + ATRA) were compared with the control group. RESULTS: When treatment groups were compared with the control group, there was a significant increase in apoptosis in all, but this was most prominent in the group treated with dexamethasone alone. The apoptosis ratios were 0.10 and 6.82% in the control and dexamethasone-only groups, respectively. We also found that there was a significant decrease in cell count, particularly in the dexamethasone only, ATRA-only, and ATRA-vitamin D(3) combination groups. CONCLUSION: ATRA, interferon-alpha, vitaminD(3) and particularly dexamethasone have significant effects on FO mouse myeloma cells resulting in a decreased cell count and an increased apoptosis ratio. This study should be repeated with human myeloma cell lines for further information. PMID- 15347913 TI - Volume flow rate evaluation in patients with obstructive arteriosclerotic disease. AB - This study was performed to quantify the volumetric impact of extracranial arteriosclerotic lesions. We investigated patients with different degrees of carotid stenosis as defined by conventional velocity-based duplex criteria and different patterns of collateralization. We studied the volume flow rate (color duplex M-mode device) in 37 patients with symmetrical internal carotid artery (ICA) plaques (<50% stenosis) and compared these data to 43 patients with a unilateral 50-74% stenosis, 75-99% stenosis (n = 73) and occlusion of the ICA (n = 37). A 75-99% stenosis caused an ipsilateral flow reduction of 36% with a wide interindividual variability, and an occlusion of 51%. Collateralization via the ophthalmic artery only caused a nonsignificant ipsilateral volume flow increase, whereas the contralateral volume flow was significantly higher in patients with a patent anterior communicating artery. The increase was more pronounced in patients with an occluded vessel. Our data demonstrate a wide diversity of the hemodynamic impact of carotid stenosis as defined by 'classical' Doppler criteria. Collateral flow reflects a dynamic quantitative process. PMID- 15347914 TI - Characterization of two transgene insertional mutations at pirouette, a mouse deafness locus. AB - The mouse mutant 'pirouette' (pi) exhibits profound hearing loss and vestibular defects due to inheritance of a recessive mutation on chromosome 5. Dysfunction has been correlated with defects during maturation of sensory cells in the inner ear. As an initial step in characterizing pirouette at the genetic level, we have localized the candidate interval to a small region on central chromosome 5 by analysis of a congenic strain of pirouette mice. This region exhibits conserved synteny with human chromosome 4 and suggests that pirouette may be a genetic model of the human nonsyndromic deafness disorder DFNB25, which has been localized to 4p15.3-q12. In addition to the original spontaneous pirouette strain, we have identified and characterized 2 additional mouse strains with allelic mutations at the same locus. Analysis of the morphology in each of the 3 pirouette alleles indicated very similar early postnatal alterations in maturation of stereocilia and suggests that the gene affected in pirouette normally plays a role in building or maintaining these structures that are critical for sensory mechanotransduction. PMID- 15347915 TI - [Reconstruction of shoulder abduction and external rotation in obstetric brachial plexus palsy]. AB - OBJECTIVES: We evaluated the results of the subscapularis and pectoralis major muscle releases and the transfer of the latissimus dorsi/teres major muscles to the rotator cuff in patients with internal rotation contractures due to obstetric brachial plexus palsy. METHODS: Seventy patients (44 boys, 26 girls; mean age 7.6 years; range 2 to 16 years) underwent transfer of the latissimus dorsi/teres major muscles to the rotator cuff. Spinal root involvement was at C5-C6 in 19 patients, at C5-C7 in 16 patients, and at C5-T1 in 35 patients. In 46 patients, the subscapularis muscle was released from the anterior surface of the scapula, and in 55 patients, the pectoralis major muscle was released by fractional tenotomy. The glenohumeral joint was evaluated by anteroposterior direct graphies and axial magnetic resonance scans. According to the Waters-Peljovich grading system, all the patients had type I or type II deformities. Pre- and postoperative range of motion values and Mallet scores were compared. The mean follow-up period was 37.9 months (range 24 to 64 months). RESULTS: The mean shoulder abduction increased to 132.6 degrees (range 90 degrees to 170 degrees; mean gain 60.3 degrees) and external rotation increased to 81.1 degrees (range 30 degrees to 100 degrees; mean gain 58.7 degrees). The mean postoperative Mallet scores for global abduction and external rotation were 3.9; hand-to-head, to mouth, and to-back scores were 3.7, 3.4, and 2.5, respectively. No serious complications were seen during the follow-up period. CONCLUSION: The results of reconstruction techniques employed in our study show satisfactory increases in shoulder abduction and external rotation in patients with a minimal glenohumeral deformity. PMID- 15347916 TI - [Early results of nerve surgery in obstetrical brachial plexus palsy]. AB - OBJECTIVES: To present surgical techniques and early postoperative results of patients who underwent nerve surgery for obstetrical brachial plexus palsy. METHODS: Twenty-four infants (12 girls, 12 boys; mean age 7.9 months; range 4 to 14 months) with obstetrical brachial plexus palsy underwent nerve repair following a surgical algorithm that showed inadequate spontaneous nerve regeneration and muscle function. Neurolysis was performed in five cases, intraplexial neurotisation in 17 cases, and extraplexial transfer of the spinal accessory nerve to the suprascapular nerve in seven cases. Seventeen patients (70%) had total palsy (C5, C6 and/or C7 rupture and C8, T1 avulsion), four patients (17%) had C5, C6 involvement, and three patients (13%) had C5-7 involvement. Pre- and postoperative evaluations were made according to the grading system of the Hospital for Sick Children (HSC). The mean follow-up period was 15.8 months (range 8 to 31 months). RESULTS: The mean HSC grades of the patients followed at least for 12 months were as follows: shoulder abduction 4, elbow flexion 4.5, wrist extension 2.3, and finger flexion 3.3. Compared to patients who underwent neurolysis alone, improved nerve regeneration was noted in patients who underwent neuroma excision and nerve grafting. No severe complications occurred postoperatively, including respiratory problems, metabolic acidosis, and hypothermia. CONCLUSION: Early diagnosis and nerve surgery in patients having insufficient muscle activity and requiring surgical intervention may prevent atrophy of the muscles and provide a more functional upper extremity. PMID- 15347917 TI - [The results of intramedullary fixation with titanium elastic nails in children with femoral fractures]. AB - OBJECTIVES: We evaluated clinical and radiologic results of intramedullary fixation with titanium elastic nails (TEN) in children with femoral fractures. METHODS: The study included 35 femoral fractures of 34 patients (20 boys, 14 girls; mean age 8.3 years; range 5 to 14 years) who were treated by intramedullary fixation with TEN. The results were evaluated according to the scoring system proposed by Flynn et al. Time to union and residual angulations were assessed on anteroposterior and mediolateral radiographs. Femoral anteversion angles and limb lengths were measured in comparison with the normal side by computed tomography (CT) in 14 patients and the results were compared using the Wilcoxon test. The mean follow-up period was 28 months (range 4 to 48 months). RESULTS: According to the criteria by Flynn et al., the results were excellent in 25 fractures (71.4%), successful in nine fractures (25.7%), and poor in one fracture (2.9%). The mean time to union was 7.4 weeks (range 5 to 12 weeks). Mediolateral and anteroposterior radiographs showed an angulation of 10 degrees or less in three fractures and of 8 degrees in one fracture, respectively. Limb length discrepancy of less than 2 cm was detected in seven patients (20.6%). Femoral anteversion angles measured by CT revealed significant retroversions on the fractured sides (p<0.01). CONCLUSION: Intramedullary fixation with TEN may be the preferred method for the treatment of femoral fractures in children aged 5 to 15 years. However, residual rotation detected by CT seems to be a technical challenge to be improved. PMID- 15347918 TI - [An evaluation of the methods used for removal of components and cement in revision hip arthroplasty]. AB - OBJECTIVES: This study was designed to evaluate the methods that we used for removal of components and cement in revision hip arthroplasty (RHA). METHODS: Thirty-seven patients (mean age 65.6 years; range 18 to 78 years) who underwent RHA were retrospectively evaluated. Arthroplasty had been cementless, hybrid, cemented, and partial in 13, 16, 5, and 3 hips, respectively. Revision included only acetabular cup in four patients, femoral component in three, and all components in the remaining patients. Of the acetabular cups revised, five were cemented, 28 were cementless (10 expanded, 18 porous-coated); of the femoral components, 22 were cemented and 11 were cementless. The mean time to revision was 7.3 years (range 1.5 to 13 years). Extended proximal femoral osteotomy (EPFO) was performed in 28 hips (84.9%), of which 10 hips (35.7%) required the use of high speed cutting heads. All the patients were evaluated before and after revision according to the criteria of Merle d'Aubigne and Postel which were modified by Charnley. The mean follow-up period was 36.3 months (range 2 to 48 months). RESULTS: There were no difficulties in the removal of cemented acetabular cups. Expanded cementless cups were readily removed by bending their expanded leaves. However, high speed cutting heads were used to resolve the bone cup integration in eight porous-coated cups (44.4%); of these, two cups (25%) even required the use of high speed metal cutting heads to split several parts apart. In one patient, a spiral fracture occurred extending to the distal end of EPFO. On final follow-ups, the results were evaluated as perfect or good in 92% of the hips. CONCLUSION: The removal of components and cement during RHA may require the use of more complex methods and device in addition to standard ones. PMID- 15347919 TI - [Necrotizing fasciitis in the lower extremity secondary to diabetic wounds]. AB - OBJECTIVES: The aim of this retrospective study was to evaluate patients who developed necrotizing fasciitis (NF) in the lower extremity secondary to diabetic wounds. METHODS: The study included sixteen patients (11 men, 5 women; mean age 63 years; range 50 to 82 years) who were treated for NF arising from diabetic wounds. Associated diseases were coronary heart disease in three patients, and chronic renal failure due to diabetic nephropathy in three patients. The patients were evaluated by physical examination, direct radiographs, blood pressure measurements, and cultures, and with respect to treatment methods and results. The mean hospitalization period was 26 days (range 5 to 59 days). RESULTS: Necrotizing fasciitis was confined to the leg in four patients, extended to the thigh in 10 patients, and was bilateral in two patients. Pain was encountered in 10 patients (68.8%). Fourteen patients (87.5%) had increased creatine kinase levels. Direct radiographs showed gas appearance in soft tissues in seven patients (43.8%). Cultures showed a single microorganism in four patients (25%), and multiple microorganisms in five patients (31.3%). Of seventeen microorganisms isolated, 10 (58.8%) were Enterobacter species. Eleven patients (68.8%) underwent above-the-knee, and three patients (18.8%) underwent below-the-knee amputations. Disarticulation of the hip was performed in four patients. The mean number of debridements was five (range 2 to 21) following the initial surgical intervention. Nine patients developed septic shock. Seven patients (43.8%) died due to multiple organ failure. Increased creatine kinase levels were significantly associated with mortality (p<0.05). CONCLUSION: Early and accurate diagnosis and prompt surgical treatment may be life-saving in diabetic patients with NF of the lower extremity. PMID- 15347920 TI - [Static and dynamic plantar pressure measurements in adolescents]. AB - OBJECTIVES: This study was designed to determine normal values of pedobarography during standing and walking in adolescents in our country and to investigate correlations between demographic data and pedobarographic values. METHODS: Fifty volunteers (25 girls, 25 boys; mean age 14 years; range 13 to 15 years) who were found to have healthy feet according to the AOFAS (American Orthopaedic Foot and Ankle Society) clinical rating system for the ankle and foot were enrolled into the study. Plantar pressures were measured during standing and walking tasks with the use of the Mini-Emed pedobarographic device. RESULTS: Static measurements showed significantly higher pedobarographic values for right medial forefoot and toes in girls, and for left midfoot in boys (p<0.05). Overall, no significant differences existed between static pressure values for the right and left feet. The mean right medial foot pressure was higher than that of the contralateral foot in girls. There were no significant differences between the right and left feet in boys. Dynamic measurements showed a significantly larger contact area of the right foot in boys, and a significantly higher maximum plantar pressure of the left medial forefoot in girls (p<0.05). The strength of the correlation of body weight and body mass index was high with maximum plantar pressures (r=0.87 and r=0.83), and moderate with contact area of the foot (r=0.63 and r=0.59) in static measurements. Body weight (r=0.64) and body mass index (r=0.54) were moderately correlated with contact area of the foot in dynamic measurements. CONCLUSION: Appreciation of normal plantar pressure values in adolescents is important in monitoring the development stages of foot, in the assessment of foot disorders, and in making proper footwear modifications in compliance with age. PMID- 15347921 TI - [Osteoid osteoma in the hand: an evaluation of eight patients]. AB - OBJECTIVES: We retrospectively evaluated patients with osteoid osteoma localized in the hand, which is a rare location for this lesion. METHODS: Eight patients (5 females, 3 males; mean age 23 years; range 13 to 31 years) underwent surgery for osteoid osteoma localized in the hand. Involvement was in the right hand in five patients, and in the left hand in three patients, being in the proximal phalanges in all the patients. The affected fingers were the fourth in five patients, and the first, second, and third in the remaining three patients, respectively. Five patients presented with pain responsive to salicylates. The mean duration of symptoms before diagnosis was 20 months (range 12 to 36 months) and the mean follow-up was 38 months (range 15 to 86 months). RESULTS: Direct radiographs enabled the diagnosis in four patients. Computed tomography was utilized in four patients, two of whom required scintigraphy and magnetic resonance imaging. Computed tomography showed intraosseous involvement in three patients, but failed to demonstrate the nidus in one patient due to extreme sclerosis. The nidus was removed with excision and curettage in all the cases, combined with cancellous autografting in five patients. Five patients had complete recovery. One patient had stiffness of the adjacent joints, which responded well to rehabilitation. Plain x-rays showed a subcortical sclerotic zone in one patient who had persistent pain for a year postoperatively. Due to inadequate excision, recurrence occurred in one patient 18 months after surgery; revisional excision and bone grafting were performed, resulting in complete recovery. CONCLUSION: Satisfactory results are obtained with excision, curettage, and bone grafting of osteoid osteomas located in the hand. PMID- 15347922 TI - [The reasons for postponement of scheduled orthopedic surgical operations and its effect on the patients' anxiety and pain levels]. AB - OBJECTIVES: This comparative-descriptive study was planned to evaluate the reasons for postponement of scheduled orthopedic surgical operations and its effect on anxiety and pain levels of patients. METHODS: The study included 100 patients (age range 21 to 56 years) who were admitted to the orthopedics department for a scheduled surgical operation in the lower extremity. Fifty patients who were subject to postponement of the operation on the scheduled day comprised the study group, and 50 patients who underwent surgery on the intended day comprised the controls. Data were collected by means of a patient questionnaire, the Spielberger State and Trait Anxiety scale, and a pain assessment scale. Evaluation of pain was made six times at regular intervals within 48 hours postoperatively. The results were compared. RESULTS: The most common reason (28%) for postponement was the presence of medical diseases on the part of the patient. Most frequently, the decision for postponement came from anesthesiologists (42%). Compared to the preoperative level, the mean state anxiety score showed a significant increase following the notification of the patients concerning the postponement (p=0.001). The number of patients who reported "disturbing pain" was at all times high in the study group, being significantly more in the second, third, and sixth evaluations. CONCLUSION: Postponed surgical operations result in an increased degree of emotional trauma and pain in patients assigned to have orthopedic surgical interventions. PMID- 15347923 TI - [Macrodystrophia lipomatosa: a case report]. AB - Macrodystrophia lipomatosa is a rare form of congenital localized gigantism. A forty-year-old male patient presented with complaints of swelling and deformity in the right toe and difficulty in wearing shoes. Conventional radiographic examination, ultrasonography, and computed tomography showed dorsal and medial deviations in the right toe, hypertrophy and degeneration in bone structure, increased density in the medullary bone, and a lipomatous mass with infiltration to the adjacent soft tissue muscles. Partial removal of the lipomatous tissues was performed. Microscopic examination of the specimen revealed fat deposits, ligamentous, and neural tissue elements in normal structure. The patient had no complaints and there was no change in the size of the lesion within a two-year follow-up period. PMID- 15347924 TI - [Diffuse synovial hemangioma of the knee: a case report]. AB - Synovial hemangioma is a rare benign tumor of joint cavities. It may be a cause of pain and recurrent joint swelling in children and young adults. Diffuse synovial hemangioma was diagnosed in a 25-year-old male patient with complaints of right knee pain and swelling of a 10-year history. Direct radiography, venography, magnetic resonance imaging, and diagnostic arthroscopy enabled the diagnosis. Surgical excision was not considered because of diffuse involvement and many pigmented areas in the skin; instead, the patient was monitored conservatively. At the end of a year, the severity of complaints and the size of the lesion did not increase. Since there is a high risk for recurrence following open synovectomy for diffuse synovial hemangiomas, conservative treatment may be an alternative approach in selected cases. PMID- 15347925 TI - [Transient osteoporosis of the hip in pregnancy: a report of three cases]. AB - Transient osteoporosis of the hip is a rare disease of unknown etiology. It is usually seen in middle-aged men and women in the third trimester of pregnancy and is associated with hip pain. It typically runs a benign course with eventual resolution of symptoms. We report three patients who presented with unilateral hip pain that started in the third trimester of pregnancy. Diagnosis of transient osteoporosis of the hip was made with the help of clinical and radiologic studies. Treatment was comprised of pain control therapy with diclofenac sodium (100 mg/day) for three months, bed rest for a month followed by mobilization with a walking aid. The follow-up period was at least 24 months. Clinical improvement occurred in the third month in one patient, and at the end of six months in the remaining patients. Complete radiologic improvement was seen at the end of 12 months. PMID- 15347926 TI - Preparation and characterization of new electrocardiogram electrodes. AB - A new generation of electrocardiogram (ECG) electrodes have been fabricated and superficially modified by a special controlled precorrosion process. The electrodes have been characterized by scanning electron microscopy (SEM), resistance-capacitance measurements (RCM) and simulated defibrillation recovery (SDR). The resulting surface of the Sn alloys was rough, and, consequently, a large active surface area was available. As a result, a higher electrocardiogram electrode performance, including low alternating current (a.c.) impedance, excellent time stability and improved SDR, was achieved. Such new design can surely find important future applications in clinical diagnosis. PMID- 15347927 TI - Mixture of epsilon-caprolactone-lactide copolymer and tricalcium phosphate: a histological and immunohistochemical study of tissue reactions. AB - In cranio-maxillofacial surgery, bone transplantation is needed for treatment of bony defects. An autograft, allograft or biomaterial can be used. Autogenous bone grafts are considered to be the best materials available, but there are some disadvantages in their use including donorsite morbidity, need for a second operative site and limited graft supply. A search for new bone-graft materials therefore remains necessary. We prepared a mixture of tricalcium phosphate (TCP), which is a resorbable, non-toxic, osteoconductive ceramic material and epsilon caprolactone-lactide copolymer P(epsilon-CL/DL-LA), a resorbable polymer, and placed it in the dermis and in mandibular bone defects in 13 rabbits. Follow-up times were two, three, seven, eight, 12, 15 and 18 weeks, tissue reactions were assessed, histologically and immunohistochemically. Times of resorption of the material from tissues were reported. We found that the mixture caused a mild inflammatory reaction when placed in bone and severe inflammation when placed in dermis. No highly fluorescent layer of tenascin or fibronectin was found surrounding the implant area. The mixture was excellent to handle and very easy to place into bone defects. The results are promising and have led us to continue development of the mixture. PMID- 15347928 TI - Tissue reactions to polyethylene implants with different surface topography. AB - This study investigates the importance of implant surface topography on soft tissue response. The tissue response in the rat abdominal wall to discs of low density polyethylene with smooth to coarse surfaces was evaluated after one, six or 12 weeks. Capsule thickness and immunohistochemical quantification of monocytes-macrophages were used as measures. The macrophage specific antibody ED1 was used for identification of newly recruited macrophages and the ED2 antibody for the mature tissue macrophages. The smoother surfaces gave a thicker capsule than the rougher surfaces, and at one week also larger total numbers of cells and ED1 positive macrophages at interface. The capsule thickness increased over time for the smooth and intermediate surface topographies. In contrast, the cell numbers generally decreased over time. In conclusion, a coarse surface elicited lesser tissue reaction compared with a smooth surface. PMID- 15347929 TI - Diamond-like carbon coatings for orthopaedic applications: an evaluation of tribological performance. AB - A detailed investigation of the tribological behaviour of vacuum arc diamond-like carbon coated Ti-6Al-4V against a medical grade ultra-high molecular weight polyethylene is conducted in this work in order to investigate the potential use of diamond-like carbon coatings for orthopaedic appplications. Lubricated and non lubricated wear experiments are performed using a standard pin-on-disc wear tester. The coefficient of friction is monitored continuously during testing and wear rate calculations are performed using surface profilometry measurements of worn disc surfaces. Sliding wear tests show the existence of two distinct friction and wear regimes distinguished by physically different mechanisms. In the first stages of wear, adhesion and abrasion are the dominant mechanisms of wear while fatigue processes are activated later in the tests. The effects of diamond-like carbon coating structure, surface roughness and lubrication on tribological behaviour are presented. Optimal process-structure-property design for vacuum arc plasma deposition is utilized in order to obtain strong adhesion to the titanium alloy substrate. Diamond-like carbon coatings significantly improve the friction and wear performance of the orthopaedic bearing pair and show exceptional promise for biomedical applications. PMID- 15347930 TI - Veneering composites - a thermoanalytical examination. AB - Differential thermal analysis and thermal gravimetric analysis were used to characterize veneering composites. Samples weighing 4-20 mg, made from the composite materials Visio-Gem (Espe, Germany), Sinfony (Espe, Germany), Artglass (Kulzer, Germany), Dentacolor (Kulzer, Germany) and Targis (Ivoclar, Liechtenstein), were examined. The samples were subjected to various thermal curing times of between 4 s and 25 min, using the relevant devices of the manufacturers. As a control group, samples of all materials were examined unreacted. In order to avoid post-curing during storage, all samples were subjected, immediately after manufacture, to the appropriate dynamic temperature programme of the thermoanalytical unit at a heating rate of 10 degrees C min(-1). The materials showed specific material characteristics which can, for instance, be used to analyse the curing behaviour of the materials. The position of the glass transition, polymerization and post-polymerization peaks at temperatures between 30 and 100 degrees C, as well as 150 and 300 degrees C, and the filler and matrix content, allow the classification of the veneering composites. PMID- 15347931 TI - Physical properties and self-setting mechanism of calcium phosphate cements from calcium bis-dihydrogenophosphate monohydrate and calcium oxide. AB - An apatitic calcium phosphate cement was developed from calcium bis-dihydro genophosphate monohydrate (or monocalcium phosphate monohydrate, MCPM) and calcium oxide (CaO). The powder had a Ca/P molar ratio of 1.67, and the liquid was either pure water or 0.25 M-1 M sodium phosphate buffer, pH 7.4. The influence of the powder-to-liquid (P/L) ratio on the setting time and the mechanical strength were studied. The best results were obtained for the 1 M phosphate buffer with a P/L ratio of 1.53; the setting time was 7 min and the compressive strength was 25 MPa after 24 h and 33 MPa after 11 d. The mechanism and kinetics of the setting reaction were investigated by X-ray diffraction, differential scanning calorimetry, 31P magic angle spinning-nuclear magnetic resonance and infrared spectrometry. The setting reaction was found to be biphasic: in the first step, during the mixing time, MCPM reacted with CaO immediately to give calcium hydrogenophosphate dihydrate (or dicalcium phosphate dihydrate, DCPD) which, in the second step, reacted more slowly with the remaining CaO to give hydroxyapatite. The conversion of the starting materials to hydroxyapatite was complete within 24 h when the liquid was water, but was slower and incomplete with the phosphate buffers. Of the starting materials, 30% remained after 3 d. PMID- 15347932 TI - Role of interconnections in porous bioceramics on bone recolonization in vitro and in vivo. AB - The interconnections in a porous biomaterial are the pathways between the pores. They conduct cells and vessels between pores. Thus they favour bone ingrowth inside ceramics. The aim of our study was to determine the effect on bone ingrowth of interconnections in two ceramics: hydroxyapatite (HA) and beta tricalcium phosphate (beta-TCP) with the same porosity of about 50% and a mean pores size of 100-300 microm and a mean interconnection size of 30-100 microm. In vitro, four discs for osteoblast culture were studied after 14 and 28 days of incubation. The results show that human osteoblasts can penetrate interconnections over 20 microm in size, and colonize and proliferate inside macropores, but the most favourable size is over 40 microm. In vivo, eight cylinders were implanted in the middle shaft of both rabbit femurs for 12 or 24 weeks. The histomorphometric results show that interconnections in porous ceramics favour bone ingrowth inside the macropores. In the HA group the rate of calcification and bone ingrowth do not differ, and chondroid tissue is observed inside pores. But in beta-TCP, the calcification rate and the bone ingrowth increased significantly. At week 12 significant correlation between new bone ingrowth and the size of the interconnections is observed between new bone ingrowth and the density of pores. In conclusion we notice that in vivo a 20 microm interconnection size only allows cell penetration and chondroid tissue formation; however the size of the interconnections must be over 50 microm to favour new bone ingrowth inside the pores. We propose the concept of "interconnection density" which expresses the quantity of links between pores of porous materials. It assures cell proliferation and differentiation with blood circulation and extracellular liquid exchange. In resorbable materials, pore density and interconnection density are more important than their size, contrary to unresorbable materials in which the sizes and the densities are equally important. PMID- 15347933 TI - Polyethylene glycol-grafted bovine pericardium: a novel hybrid tissue resistant to calcification. AB - Calcification is a frequent cause of the clinical failure of bioprosthetic heart valves fabricated from glutaraldehyde pretreated bovine pericardium (GATBP). An investigation was made of the grafting of different molecular weight polyethylene glycol (PEG 600, 1500, 4000 and 6000) via glutaraldehyde (GA) linkages to bovine pericardium (BP) and of their stability and calcification. The process of the calcification profile was studied by in vitro experiments via incubating pericardial samples in a metastable solution of calcium phosphate. Calcification of bovine pericardium grafted with PEG 6000 was significantly decreased compared to low molecular weight PEG grafts or Sodium dodecyl sulphate- (SDS) and GA treated tissues. The mechanical properties of these modified tissues after enzyme (Trypsin) digestion and calcification were investigated. The biocompatibility aspects of grafted tissues were also established by monitoring the platelet adhesion, octane contact angle and water of hydration. PEG 6000-grafted tissues retained the maximum strength in trypsin buffer and calcium phosphate solutions. Scanning electron micrographs revealed that the PEG-grafted bovine pericardium had substantially inhibited the platelet-surface attachment and their spreading. It is conceivable that high molecular weight polyethylene glycol-grafted pericardium (a hybrid tissue) may be a suitable calcium-resistant material for developing prosthetic valves due to their stability and biocompatibility. PMID- 15347934 TI - Biomaterials science: prospects for the new millenium? PMID- 15347935 TI - Influence of sodium oxide content on bioactive glass properties. AB - The rate of in vivo degradation and level of bioactivity of bioactive glasses are composition dependent [1]. By altering bioactive glass composition, the rate of resorption can be controlled. The network connectivity of a glass can be used to predict various physical properties of the glass including its solubility and, hence, its bioactivity [2]. Glass solubility increases as network connectivity is reduced. Glasses in the soda-lime phosphosilicate system were studied. The initial choice of composition was based on phosphate content and low network connectivity. A systematic substitution of calcium oxide for sodium oxide on a molar basis was made in order to examine the influence of sodium oxide content on the glass properties while keeping the network connectivity constant. The glass transition temperature and the peak crystallization temperature were seen to decrease linearly with increasing sodium oxide content. Thermal expansion coefficient and glass density were also seen to be related to sodium oxide content. Preliminary in vitro biocompatibility studies revealed that the glasses of higher sodium oxide content were associated with a cytotoxic response. The measurement of media pH indicated that this cytotoxic effect was due to ion exchange reactions at the glass surface. PMID- 15347936 TI - Immobilization of a biologically active coating on a hydrophobic L-lactide epsilon-caprolactone copolymer. AB - The electron beam radiation induced grafting method was used to attach a reactive polyacrylamide (PAA) layer (20 wt %) on the surface of a biodegradable poly-L lactide-co-epsilon-caprolactone (PLLA-co-CL). The biocompatibility of graft polymer obtained was studied by cytotoxicity test and no signs of toxicity were observed. Heparin and sol-gel-produced silica-gel coatings were successfully attached on the top of the polymeric material produced. The amount of heparin immobilized directly on the surface can be controlled by reaction conditions: reaction time, temperature and pH of the incubation solution. By using acidic conditions, up to 98 microg cm(-2) of heparin was immobilized on the surface. The sol-gel-produced silica-gel layer formed by dipping technique was 30 microm thick and the cracking of the layer was minimal after bending several times to 90 degrees. PMID- 15347937 TI - Bioabsorbable polymer plates coated with bioactive glass spheres. AB - In order to provide bioabsorbable self-reinforced poly (L/DL) lactide (P(L/DL)LA) 70 : 30 plates with osteoconductive bioactivity, spheres (125-250 microm) of a bioactive glass 13-93 were implanted onto a polymer plate preform by pressing. With appropriate pressing parameters glass spheres were firmly attached onto the polymer plate. The top of the glass spheres remained exposed. The bioactivity of the coated plates was examined in vitro with immersion in phosphate-buffered saline (PBS). Apatite was observed to precipitate on exposed glass sphere surfaces and the whole polymer surface within the first three days. PMID- 15347938 TI - Polyurethane-maleamides for cardiovascular applications: synthesis and properties. AB - Several polyurethane-maleamides (PUMAs) containing polyether or polycarbonate soft segments, and aromatic or aliphatic hard segments were synthesized by solution or bulk polymerization, using maleic acid (MA) or a mixture of MA and butanediol as chain extenders. Using this process, activated double bonds are introduced into the polymer chains and the base polyurethanes may undergo further modification via specific grafting, thus improving their tissue compatibility. PUMAs chemicophysical properties were evaluated by gel permeation chromatography (GPC), intrinsic viscosity analyses, differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR) and tensile mechanical tests. Polycarbonate diol (PCU)-based PUMAs showed higher molecular weights than polyether diol (PEU)-based ones. The use of butanediol in mixture with maleic acid led to an increase of molecular weights. FT-IR confirmed the presence of the bands related to the amide groups and to the conjugated double bond, yet more evident for the polymer obtained in solution. The higher crystallinity shown by this polymer was also indicative of a better phase separation. All the PCU-PUMAs exhibited similar tensile properties with a higher stiffness than PEU-PUMAs. Among the PEU-PUMAs, the highest tensile properties were shown by the polymer obtained in solution, and by the one derived from a mixture of maleic acid and butanediol. PMID- 15347939 TI - Zirconia-toughened hydroxyapatite ceramic obtained by wet sintering. AB - A toughened hydroxyapatite (HA) ceramic has been obtained through the incorporation of magnesia partially stabilized zirconia (Mg-PSZ) under uniaxial pressing and sintering in wet oxygen at 1250 degrees C for 4 h. The powder X-ray diffraction (XRD) patterns and infrared spectra (FT-IR) show that HA is the only calcium phosphate phase present. The composite (MgPSZ-HA) has a density of 94% the theoretical value. The bending strength and the fracture toughness are around 50% higher for Mg-PSZ reinforced than for HA. The grain size and the fracture surface were studied by scanning electron microscopy (SEM). The influence of the Mg-PSZ particles on the fracture mechanism of the HA ceramic is discussed. PMID- 15347940 TI - Neuronal networks in vitro: formation and organization on biofunctionalized surfaces. AB - Receptor-mediated recognition of substrate molecules is a prerequisite for nerve cells in order to find their target structures in vivo and leads to formation of neuronal connections and networks. In order to study these mechanisms under in vitro conditions, we cultured embryonic hippocampal neurons or neuronal cell lines, SH-SY5Y and PCC7-PCC7-Mz1, onto biofunctionalized surfaces. Micropatterning on polymer surfaces, glass- and silicone-oxide-based chip materials was performed in a micrometer range by microcontact printing using polydimethylsiloxane (PDMS) stamps. Hippocampal neurons were found to form networks on chip surfaces under serum-free conditions and remained functional for more than a week. Human neuroblastoma cells SH-SY5Y as well as PCC7-Mz1 stem cells were found to follow microcontact printed pattern on polystyrene surfaces. Both cell lines showed neuronal marker expression and were cultured for up to 7 days with serum containing culture medium. Widths of 3-5 microm of coating lines were found to enhance single cell spreading along the pattern. The techniques described in this study may be useful in promoting nerve cell regeneration and organization following transection due to trauma or surgery. The neuronal alignment and network formation in vitro may furthermore serve as a model system in the field of biosensors. PMID- 15347941 TI - Analysis of the structural changes of a phosphate glass during its dissolution in simulated body fluid. AB - The structural changes of two calcium-sodium metaphosphate glasses during its dissolution in simulated body fluid (SBF) have been analyzed by Fourier transform Raman spectroscopy (FT-Raman), scanning electron microscopy (SEM) and environmental scanning electron microscopy (ESEM). The results showed that no structural changes could be detected during the first week of dissolution. However, after approximately 4 weeks of dissolution the analysis of the glass surface revealed the presence of a thin orthophosphate layer. The elemental analysis of this layer by X-ray dispersive energy showed the presence of calcium phosphate ions, while almost no sodium was detected. The observation of the glasses by ESEM showed the formation of small crystals when the water vapor pressure of the chamber was decreased. This indicates that the layer analyzed by SEM and FT-Raman was related to a hydrated calcium phosphate layer. PMID- 15347942 TI - Improvement of the mechanical properties of acrylic bone cements by substitution of the radio-opaque agent. AB - Acrylic bone cements become radio-opaque by the addition of an inorganic compound, commonly BaSO4 or ZrO2. However, the use of these additives has some negative effects such as loss of mechanical properties, risk of release and bone resorption. The use of the monomer 2,5-diiodo-8-quinolyl methacrylate (IHQM), which shows adequate polymerization and radio-opacity properties, is proposed as a new X-ray opaque, methacrylate iodine-containing agent. The aim of this study is to determine the effect of this new radio-opaque agent on the mechanical properties of acrylic bone cements. The addition of the iodine-containing methacrylate provides a statistically significant increase in the tensile strength, fracture toughness and ductility, with respect to the barium sulphate containing cement. This effect can be attributed to the fact that the use of a radio-opaque monomer eliminates the porosity associated with the barium sulfate particles, which show no adhesion to the matrix. However, some reinforcing effect must also be attributed to the iodine-containing monomer, since the tensile and fracture toughness values reached are even higher than those shown by the radiolucent cement. PMID- 15347943 TI - Biomechanical evaluation of cell-loaded and cell-free hydroxyapatite implants for the reconstruction of segmental bone defects. AB - Porous hydroxyapatite (HA) scaffoldings are currently used in tissue engineering for bone reconstruction. When this osteoconductive biomaterial is combined with osteoprogenitor cells, it acquires osteoinductive features which accelerate and improve bone formation in vivo. The aim of our study was to assess the mechanical properties of HA-bone complexes undergoing indentation tests, and relate stiffness to composition and structure as examined by micro X-ray. To this purpose, 35-mm tibia diaphyseal resections were performed in sheep. Gaps were filled using porous HA cylinders. Implants were loaded with autologous bone marrow stromal cells (BMSC); cell-free cylinders were used as control. After 8 weeks, bone tissue was found within the internal macropores of cell-loaded HA carriers, and in control implants, bone formation was mostly limited to the outer surface. As assessed by indentation testing the stiffness values of bone-HA composites were halfway between those of HA scaffoldings and tibia bone. Cell loaded implants were stiffer than cell-free ones. In a cell-loaded implant we also analyzed the variation of stiffness along the main axis of the tibia. PMID- 15347944 TI - Polymeric prodrugs of antibiotics with improved efficiency. AB - Macromolecular prodrugs of the antibiotic norfloxacin were prepared by coupling the drug via a peptide spacer onto a mannosylated dextran. The tetrapeptide gly phe-gly-gly-gly-OMe was selected as substrate for lysosomal enzymes. The drug was coupled on the alpha-C of the terminal glycine. In vitro degradation studies demonstrated the release of the parent drug in the presence of cathepsin B. In vivo experiments on mice showed a promising therapeutic effect. PMID- 15347945 TI - Chemical micropatterning of polymeric cell culture substrates using low-pressure hydrogen gas discharge plasmas. AB - Micropatterned cell cultures will allow a new quality of bioartificial systems. Here, an approach to chemical micropatterning of polymer substrates is presented, which is completely based on low pressure gas discharge processes. Well expressed micropatterned cell cultures on polystyrene and poly (ether ether ketone) were obtained with many different cell types. No impairment of typical cell behavior was observed. PMID- 15347946 TI - Determination of mechanical properties of impacted human morsellized cancellous allografts for revision joint arthroplasty. AB - This paper deals with the characterization of mechanical properties of impacted morsellized cancellous allograft (IMCA) produced by dynamic compaction of allograft femoral heads ground by commercially available bone mills, i.e. rotating rasp and reciprocating type bone mills. Various ranges and profiles of particle size in the graft aggregates were obtained using these bone mills, and the effect of number of compaction as well as the distribution of particle sizes on the mechanical properties of IMCA under quasistatic compression and shear loading conditions was discussed. The morsellized cancellous allograft prepared by the reciprocating type bone mill showed a broad distribution of particle sizes, and gave IMCA superior mechanical properties to the graft with a more uniform size distribution, or prepared by the rotating rasp type bone mills. The increase of number of compaction also improved the mechanical properties of IMCA in compression. PMID- 15347947 TI - Image and fractal analysis of osteoblastic cells in viscous media. AB - The aim of the present study was to determine whether osteoblastic morphology was affected by the viscosity of culture media and whether any morphological differences could be readily quantified. A cytochemical stain was used for alkaline phosphatase and a combination of image and fractal analysis, utilising seven morphological parameters to assess the morphology of osteoblasts. Using regression analysis it was determined that as the viscosity of the culture media increased the area of the cells significantly decreased and the fractal dimension of the cell profiles significantly increased. A discriminant function analysis was used to examine whether cell populations could be classified according to culturing time or the viscosity of culturing media based on the seven morphological parameters. It was determined that the cells could be classified up to 93% correct according to the viscosity of the media they were cultured in and up to 93.5% correct according to the culturing time. This study demonstrated that viscous media affects the morphology of osteoblastic cells and that discriminant function analysis can be used to classify these cells based on their morphological parameters. PMID- 15347948 TI - Bone engineering on the basis of periosteal cells cultured in polymer fleeces. AB - Large bone defects caused by severe trauma, infection or tumor resection are still a major challenge for orthopaedic surgery. The key concept for successful bone regeneration consists of combining the osteoinductive effect of osteogenic cells with a suitable carrier structure to promote osteoblastic differentiation and optimal matrix production. Therefore, periosteal cells cultured in polyglycolic-polylactid acid (PGLA) fleeces were investigated for their osteogenic differentiation and used to repair critical size bone defects in a rabbit model. Periosteal cells were isolated from New Zealand White rabbits and expanded in vitro. Osteogenic differentiation was investigated by analysis of alkaline phosphatase and osteocalcin production in vitro depending on culture conditions and passage number. Cells were seeded into PGLA fleeces. After further cultivation, tissue constructs were examined histologically and by immunohistochemistry for cell distribution and osteogenic differentiation. These constructs of defined size were used to repair critical size calvarial defects (group I) in rabbits compared to a defect repair with polymers only (group II) or to untreated defects (group III). Bone healing was evaluated after 4 weeks by radiodensitometry and a special histological scoring system. For early evaluation, radiodensitometry was not sensitive enough to detect differences in calcification. However, on histologic examination the group with cell/fleece constructs revealed intense formation of uncalcified bone. The mean defect closure of the experimental group I was 65%, compared to control groups II and III with 31% and 22%, respectively. The established methods of 3-D-cell culture and ex-vivo transplant assessment proved to be a valuable tool for quality assurance. The results demonstrate that the combination of periosteal cells and polymer fleeces is a tissue engineering approach, which may have clinical applications in various fields of reconstructive surgery. PMID- 15347949 TI - Static and dynamic fibroblast seeding and cultivation in porous PEO/PBT scaffolds. AB - The present study aims at optimizing dermal fibroblast seeding and cultivation in Polyactive scaffolds in order to limit the biopsy size needed for autologous treatment of full-thickness skin defects and chronic wounds. Three methods for seeding and cultivation of fibroblasts in porous scaffolds were compared: dynamic seeding followed by static cultivation (DS), static seeding followed by static cultivation (SS) and dynamic seeding followed by dynamic cultivation (DD). Human dermal fibroblasts isolated from cultured explants were seeded in porous PEO/PBT (Polyactive) scaffolds. Samples were taken from 6 h to 21 days post-seeding for both histological analysis (cell distribution and extracellular matrix (ECM) formation), and quantitative cell number assay. The seeding efficiency 24 h post seeding was 76% (+/-3.6%) for dynamically seeded matrices, whereas it was only 30% (+/-5%) for statically seeded matrices (p<0.001). ECM formation was abundant in DS samples already at day 10, while even after 21 days ECM formation was less pronounced in SS samples. Surprisingly, cells detached from DD samples as aggregates, starting from day 10. Cell numbers as assayed quantitatively correlated with the histological results. At all timepoints cell numbers found for DS samples were significantly higher as compared to SS samples. At day 21, DS samples contained approximately twofold more cells as compared to SS and DD samples and comprised ECM consisting of collagen types I and III. Our results indicate that the combination of dynamic seeding and static cultivation assures efficient utilization of isolated fibroblasts and improved neodermis formation, thereby allowing a reduction in the skin biopsy size needed for the engineering of living skin substitute. PMID- 15347950 TI - Measurement of non-linear microcrack accumulation rates in polymethylmethacrylate bone cement under cyclic loading. AB - Damage accumulation in the cement mantle used to fixate bone prosthesis is one failure scenario for joint reconstruction. It can be described as the phenomenon of numerous microcracks initiating and propagating within the material. Microcracks grow in the cement mantle causing it to gradually lose its mechanical integrity, leading to loosening of the prosthesis. In this study microcracking within acrylic bone cement was quantified over the course of a fatigue test. Identification of new cracks and the growth of pre-existing cracks was monitored at intervals during fatigue testing of five specimens at a mean cyclic stress of 7.5 MPa. Given these measurements, an average damage evolution curve was derived for acrylic bone cement. It was observed that the initiation sites for microcracks were the pore perimeters; therefore, the number of microcracks present in a sample is dependent on porosity. Variability was found within the results and the majority of the variability was accounted for by the difference in the porosity of each sample. Results have identified non-linear damage evolution. On a simplified level a power law equation can be used to describe the damage evolution process. PMID- 15347951 TI - An in vivo comparative study of the e-polytetrafluoroethylene vascular prostheses: Vitaflon and Gore-Tex. AB - A comparative study was performed in order to validate new Russian e-PTFE vascular prostheses Vitaflon (St. Petersburg, Russia). The Gore-Tex prostheses were chosen as a referential model. The prostheses were implanted in the venous and arterial positions in 13 dog experiments. After the implantation time was over a comprehensive histological and histochemical examination of excized specimens was performed. It was demonstrated that there is no difference in healing and functional properties between the two studied prostheses. PMID- 15347952 TI - Neurite outgrowth on microstructured surfaces functionalized by a neural adhesion protein. AB - Designed networks of neurons are potentially very useful to investigate neural activities. Using photolithography microgrooves suited in size for single neurons have been produced on glass chips. Two conducting gold lanes ending in each microgroove allow extracelluar stimulation of the neurons and recording of their activity. A cell adhesive surface was created by functionalization of glass with the adhesion peptide RGDC. In addition, in order to optimize the contact of the neuronal cell membrane to the electrode surface axonin-1, a specific neural adhesion protein was used. A recombinant form of axonin-1 was produced and immobilized in a correct orientation on protected gold surfaces through a C terminal cysteine residue. Neurite outgrowth of neurons cultured on chips derivatized with RGDC or axonin-1 were compared. The developed materials and methods represent a first step towards establishing designed functionalized glass surfaces for neurophysiological investigations. PMID- 15347953 TI - In vitro evaluation of a new polymethylmethacrylate cement reinforced with hydroxyapatite. AB - The nature of the orthopedic implant surface affects the interaction between cells and subsequent bone formation. The bone/cement interface in cement-held prostheses is considered to be the main cause of fracture leading to implant revision. It is thought that the introduction of a bioactive phase, such as hydroxyapatite (HA), to cement may permit a stronger implant by encouraging direct bone apposition rather than encapsulation of the implant by fibrous tissue. Thus, a poly(methylmethacrylate) (PMMA) cement incorporating 17.5% HA by weight has been investigated. In this study, in order to analyze the interaction at the cellular level, the in vitro biological response of the HA/PMMA to a similar PMMA without HA incorporation has been studied. Primary human osteoblast like cells (HOB) were used as they are a model of the cell type the cements might encounter in vivo. Cell proliferation and growth were assessed by measurement of total cellular DNA and tritiated thymidine ([3H]-TdR) incorporation. Alkaline phosphatase (ALP) production was measured as an indicator of HOB phenotype upon the cements. The results showed that HA/PMMA was a better substrate for HOB cells, resulting in increased proliferation and ALP activity. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed that HOB cells cultured on the HA-filled PMMA preferentially anchored to HA particles exposed at the cement surface, with a close intimacy observed between HA and HOB cells. PMID- 15347954 TI - Differential gene expression after implantation of biomaterials into rat gastrointestine. AB - Two biodegradable materials, polyglycolide-copoly(L)-lactide/poly-dioxanone composite and poly(3-hydroxybutyrate), have been employed for surgery in the gastrointestine of laboratory rats. The tissue response was analyzed at distinct time intervals up to 2 months by differential mRNA display. Two specific PCR fragments were transiently present 7 and 14 days after contact with poly(3 hydroxybutyrate). Both fragments represent distinct rat lipases which might play a role in poly(3-hydroxybutyrate) degradation. PMID- 15347955 TI - A novel ex vivo model for investigation of fluid displacements in bone after endoprosthesis implantation. AB - Tissue perfusion and mass transport in the vicinity of implant surfaces prior to integration or bonding may play a crucial role in modulating cellular activities associated with bone remodeling, in particular, at early stages of the integration process. Furthermore, fluid displacements have been postulated to transduct mechanical stress signals to bone cells via loading-dependent flow of interstitial fluid through the lacunocanalicular network of bone. Thus, an understanding and new possibilities for influencing these processes may be of great importance for implant success. An ex vivo model was developed and validated for investigation of fluid displacements in bone after endoprosthesis implantation. This model serves to explicate the effects of surgical intervention as well as mechanical loading of the implant-bone construct on load-induced fluid flow in the vicinity of the implant. Using this model, we intend to quantify perfusion and extravascular flow dynamics in the vicinity of implants and define optimal conditions for enhancing molecular transport of osteotropic agents from the implant surface to apposing bone as well as from the blood supply to the implant surface. Furthermore, the elucidation of main transport pathways may help in understanding the distribution of wear particles in bone surrounding implant, a process which has been postulated to cause osteolysis and implant loosening. PMID- 15347956 TI - In vitro assessment of endothelial cell adhesion mechanism on vascular patches. AB - Endothelial cell (EC) seeding of small caliber vascular grafts prior to their implantation has proved to significantly improve long-term patency in humans. We have previously demonstrated that a monolayer of EC could be obtained on type I collagen-coated knitted ultrathin polyster grafts (InterVascular, La Ciotat, France). Thus, the aim of the present work was to understand the nature of cell adhesion mechanisms involved in the cell /biomaterial interface, using HemaCarotid (InterVascular) patches made of type I collagen-coated knitted ultrathin polyster (type I collagen is used to coat patches to attain low permeability). By means of quantitative attachment tests, adhesion blocking assays, RT-PCR for the expression of beta1 integrin mRNA, indirect immunofluorescence with antivinculin antibody, we were able to show that EC are able to adhere to such surfaces by the means (non-unique) of cell surface receptors of the beta1 integrin group. However, the latter are probably downregulated at the cell/biomaterial interface. PMID- 15347957 TI - Histological aspects in bone regeneration of an association with porous hydroxyapatite and bone marrow cells. AB - The osteogenic potential of an association of two kinds of hydroxyapatite (HA) porous ceramics with autologous bone marrow cells cultured with or without dexamethasone (10(-8)M) addition in the culture medium and non-cultured rabbit marrow stromal stem cells (MSCs) was tested after 4 weeks of implantation in the dorsal muscles of spine in rabbit. A significantly higher number of rabbits with implants containing bone tissue inside pores were obtained with 10(7) cells ml( 1) cultured treated with Dex. In the HA porous ceramic using naphtalen as porogen agent, the bone recolonization remains only at the periphery of implants and in the second row of pores, while in the HA porous ceramic using polymethacrylate (PMMA) microbeads as porogen agent, the bone recolonization is observed in the depth of implants. In the PMMA HA group, the Kruskal-Wallis variance analysis between the rabbits is significantly different with the percentage of number of occupied pores and occupied pores with bone tissue is different (p<0.05). PMID- 15347958 TI - Human osteoblast adhesion on titanium alloy, stainless steel, glass and plastic substrates with same surface topography. AB - Osteoblast adhesion on materials will depend on the surface aspects of materials which may be described according to their surface chemistry, surface topography or surface energy. To separate the effects of roughness and composition of materials on osteoblast response, we chose to compare substrates with various surface composition but with the same smooth surface. Ti6Al4V alloy, stainless steel, glass and standard tissue culture polystyrene were tested. Adhesion was evaluated using specific antibodies against adhesion proteins and by a quantitative cell detachment assay. After 1, 7 and 14 days, cells expressed extracellularly fibronectin fibers, and intracellularly type I collagen and osteopontin. Vinculin-labeled focal contacts were visible on all materials but were more frequent on glass and stainless steel surfaces. beta_1-integrin subunit labeled patches were visible on all surfaces at each delay. The quantitative cell detachment assay showed few differences between materials. Adhesion was higher on metallic substrates although cell proliferation was higher on glass and stainless steel compared to tissue culture polystyrene and Ti6Al4V alloy. Substrates with various surface composition but with the same surface topography did not induce significant differences of adhesion although cell proliferation was variable. PMID- 15347959 TI - Biphasic synthetic bone substitute use in orthopaedic and trauma surgery: clinical, radiological and histological results. AB - Searching for an alternative to bone grafts in orthopedic and trauma surgery, two biphasic synthetic calcium phosphate ceramics BCP are made: the first, Eurocer 400, in granule form with a high interconnected porosity for void-filling, and the second, Eurocer 200, available in different shapes, with a good mechanical resistance in compression. Two hundred cases are reported with more than a six month follow-up. The first 72 cases relating to hip arthroplasty revision surgery (29 involving acetabular and 43 femoral stem loosening) are exposed with some technical details. The next 71 cases concerning trauma and sequels are displayed with technical particulars. The remaining 57 other cold orthopedic indications are then enumerated. The very good biocompatibility of these ceramics is confirmed. Radiological incorporation is quickly seen in all the cases, faster with the filling substitute than with the second one, which, however, presents no mechanical failures when classical technical principles are respected. Finally, some histological studies are presented; the ceramics are progressively resorbed and bone reconstruction in and close to the substitutes is noticed. Therefore, the use of biophasic ceramics in almost all orthopedic and trauma surgery is recommended. PMID- 15347960 TI - Surface modification tailors the characteristics of biomimetic coatings nucleated on starch-based polymers. AB - This work describes the influence of surface pretreatments over the nucleation and growth of an apatite layer, formed by a biomimetic process, on which a bioactive glass is used as a precursor of the calcium-phosphate (Ca-P) formation on the materials surface. SEVA-C, a corn starch-based biodegradable blend, was used as substrate. The surfaces were pretreated during various periods by: (i) physical methods, namely ultraviolet radiation (u.v.), and over exposure to ethylene oxide sterilization (EtO); and (ii) chemical methods, namely potassium hydroxide (KOH) and acetic anhydride (CH3CO)2 etchings. The surface modifications, performed before the production of the biomimetic coatings, resulted in a faster formation of Ca-P nuclei during the first stages of SBF immersion, particularly in the case of the KOH etching. In this case, it was possible to observe a decrease in the average surface roughness, as measured by laser profilometry, and an increase of the hydrophilicity of the material, which was evident from a clear increment in the water-uptake ability and quantified by contact angle measurements. With this treatment it was possible not only to reduce the induction period for the formation of a well defined and dense apatite like layer, as observed by scanning electron microscopy (SEM), but also to improve the adhesion of the Ca-P layer to the substrate, as confirmed by the adhesion strength tests. For all the studied pre-treatments, the composition of the films, analyzed by energy dispersive spectroscopy (EDS) and identified by thin-film X-ray diffraction (TF-XRD), seems to be very similar to that of human bone apatites. PMID- 15347961 TI - Induced tissue integration of bone implants by coating with bone selective RGD peptides in vitro and in vivo studies. AB - The optimal function of medical implant materials used in tissue substitution is often limited due to its healing properties. This effect is linked to reduced interactions of the implants with the surrounding tissue. Implant surfaces biologically functionalized with arginine-glycine-aspartic acid (RGD) peptides, a class of cellular adhesion factors, are described in this paper. The RGD-peptides are either bound via bovine serum albumin linking on culture plastic dishes as a model surface or via acrylic acid coupling on PMMA surface as a potential implant material. Resulting functionalized surfaces aquire the capability to bind cultured osteoblasts in high levels and show high proliferation rates in vitro. These results are observed for osteoblast cultures as well as from different species with different preparation procedures. A critical minimum distance between the bioactive portion of the RGD-peptides and the implant surface of 3.0 3.5 nm is crucial for the induction of an optimum cell binding process. In vivo animal studies in the rabbit show that newly formed bone tissue generated a direct contact with the RGD-peptide coated implants. In contrast uncoated implants are separated from newly formed bone tissue by a fibrous tissue layer thereby preventing the formation of a direct implant-bone bonding. PMID- 15347962 TI - A combined finite element method and continuum damage mechanics approach to simulate the in vitro fatigue behavior of human cortical bone. AB - The fatigue of bone, in particular the associated modulus degradation and accumulation of permanent strain, has been implicated as the cause of femoral neck fractures and the migration of total joint replacements. The objective of this study was to develop a technique to simulate the tensile fatigue behavior of human cortical bone. A combined continuum damage mechanics (CDM) and finite element analysis (FEA) approach was used to predict the number of cycles to failure, modulus degradation and accumulation of permanent strain of human cortical bone specimens. The simulation of fatigue testing of eight dumb-bell specimens of cortical bone were performed and the predictions compared with existing experimental data. The predictions from the finite element models were in close agreement with the experimental data. The models predicted similar development of modulus degradation and permanent strain as observed in the experimental tests. The technique is capable of predicting the accumulation of permanent strain without the need for simulating every single load step. These findings suggest that the complex fatigue behavior of human cortical bone can be simulated using the described approach and forms the first step for simulating the more complex mechanisms associated with femoral neck fractures and implant migration. PMID- 15347963 TI - Titanium levels in rats implanted with Ti6Al4V treated samples in the absence of wear. AB - The effect of implantation time and implant nitriding on titanium ion concentration in several tissues of rats carrying Ti6Al4V implants was studied by means of inductively coupled plasma-mass spectroscopy (ICP-MS). Histological studies were also performed in order to check for tissue degeneration due to the Ti6Al4V implantation. The animals were divided into four groups: one received Ti6Al4V implants, the second received nitrided Ti6Al4V implants, the third group received nitrided and descaled Ti6Al4V implants and the last one was the control group. Half the animals of the implanted groups received the Ti6Al4V implant for 30 days, while the other half received the implant for 120 days. Spleen, muscle, kidney, lung, brain and bone samples were retrieved from these rats as well as the control group. Ion concentration measures did not show significant differences between control and implanted rats for the studied period of time, although histological studies showed minor differences, especially on liver tissue samples. PMID- 15347964 TI - Initial adhesion and surface growth of Pseudomonas aeruginosa on negatively and positively charged poly(methacrylates). AB - The infection risk of biomaterial implants is determined by an interplay of bacterial adhesion and surface growth of the adhering organisms. In this study, we compared initial adhesion and surface growth of Pseudomonas aeruginosa AK1 (zeta potential -7 mV) on negatively charged (PMMA/MAA, zeta potential -18 mV) and positively charged (PMMA/TMAEMA-Cl, zeta-potential +12 mV) methacrylate copolymers in situ in a parallel plate flow chamber. Initial adhesion was measured using phosphate-buffered saline and subsequent surface growth of the adhering bacteria using nutrient broth as growth medium. Initial adhesion was twice as fast on the positively charged methacrylate than on the negatively charged copolymer. Surface growth, however, was absent on the positively charged copolymer, while on the negatively charged methacrylate the number of bacteria increased exponentially during surface growth with a generation time of 32 min. From the results of this study it can be concluded that positively charged biomaterial surfaces might show reduced risks of biomaterials-centred infections, despite being more adhesive. PMID- 15347965 TI - Dynamic mechanical properties of hydroxyapatite-reinforced and porous starch based degradable biomaterials. AB - It has been shown that blends of starch with a poly(ethylene-vinyl-alcohol) copolymer, EVOH, designated as SEVA-C, present an interesting combination of mechanical, degradation and biocompatible properties, specially when filled with hydroxyapatite (HA). Consequently, they may find a range of applications in the biomaterials field. This work evaluated the influence of HA fillers and of blowing agents (used to produce porous architectures) over the viscoelastic properties of SEVA-C polymers, as seen by dynamic mechanical analysis (DMA), in order to speculate on their performances when withstanding cyclic loading in the body. The composite materials presented a promising performance under dynamic mechanical solicitation conditions. Two relaxations were found being attributed to the starch and EVOH phases. The EVOH relaxation process may be very useful in vivo improving the implants performance under cyclic loading. DMA results also showed that it is possible to produce SEVA-C compact surface/porous core architectures with a mechanical performance similar to that of SEVA-C dense materials. This may allow for the use of these materials as bone replacements or scaffolds that must withstand loads when implanted. PMID- 15347966 TI - Analysis of push-out test data based on interfacial fracture energy. AB - Push-out testing is frequently used to assess the interfacial shear strength developed at a bone-biomaterial interface during in vivo experiments. The aim of the present research was to assess the in vivo performance of a novel substrate/coating combination and to introduce a more rigorous fracture mechanics analysis of the push-out test data. An adhesively bonded hydroxyapatite (HA), and a Ti-6Al-4V alloy plasma sprayed with HA, were implanted in female New Zealand white rabbits for up to 6 months in duration. After death, push-out tests were carried out and the shear strength was calculated in the conventional way, together with microscopical examination of crack paths. A finite element model was drawn up representing four potential failure mechanisms. The measured "failure shear strengths" in conventional analysis were approximately equal for the two coatings. However, JC at failure calculated from the model was 210 J m( 2) at the novel adhesively bonded HA/bone interface and 5 J m(-2) at a conventional titanium/plasma-sprayed HA interface. The conventional shear strength approach is strongly test dependent, and we believe that the fracture energy approach represents a more rigorous analysis of the real failure criterion in the implant/host tissue structure. PMID- 15347967 TI - Effects of surface-treated cpTi and Ti6Al4V alloy on the initial attachment of human osteoblast cells. AB - This study concerns the effect of simple surface treatments on the nature of the oxide layer, of commercially pure titanium (cpTi) and Ti6Al4V alloy substrates and their effect on human osteoblast cells (HOBS). After treatment the surfaces were analyzed by X-ray photoelectron spectroscopy (XPS) in order to identify the surface groups responsible for the cell attachment process. The assessment of cell attachment was monitored by the Alamar blue assay (AB), measuring cell activity, in three types of media: phosphate-buffered saline (PBS), serum containing and serum-free Dulbecco's modified Eagle's cell culture medium (SER+ and SERF respectively). XPS analysis of the treated surfaces revealed consistent peaks representative of TiO2 on all surfaces and Ti(0) and Ti2O3 on the non-heat treated surfaces. The cell activity assays indicated that there were no significant differences in cellular activity caused by surface treatments, but the cellular activity compared between the three types of medium was greatest in the PBS over the initial stages of attachment. PMID- 15347968 TI - New resorbable polymeric systems with antithrombogenic activity. AB - The synthesis and application as resorbable coatings of vascular grafts of a new polyacrylic derivative of Triflusal (2-acetyloxy-4-trifluoromethyl)benzoic acid, a commercial drug with antithrombogenic properties, are described. The high molecular-weight polyacrylic system is rather stable in physiological conditions and provides a chemical support for the slow release of the pharmacologically active compound, Triflusal, or its main metabolite (2-hydroxy-4 trifluoromethyl)benzoic acid (HTB). Experiments of deposition and retention of platelets in static basal conditions using plasma-rich medium from blood of sheep, seem to indicate that the polymeric coating of the polyacrylic derivative of Triflusal improves the antiaggregating character for platelets of the surface of small-diameter vascular grafts without the application of other antithrombogenic drugs. PMID- 15347969 TI - In situ localization of apoptotic changes in the interface membrane of aseptically loosened orthopaedic implants. AB - Twenty specimens of bone-implant interface membrane from THR/TKR were used for in situ localization of apoptotic changes. A panel of antibodies was used to label leukocyte antigens (CD68 and CD3) cytokines (IL-1alpha and IL-1beta) and apoptosis inhibiting and promoting proteins (bcl-2 and bax) by means of immunohistochemical techniques. A DNA fragment test on the tissue sections was also carried out to confirm actual cell death using the enzyme terminal deoxy nucleotidyl transferase (TdT) to incorporate biotinylated nucleotide with the 3' OH DNA ends. Leukocyte antigen staining showed that there were large numbers of CD68 positive macrophages as well as multinucleate giant cells (MNGC) but that CD3 positive lymphocytes were also present in the interface membrane. The leukocyte surface antigen staining pattern corresponded to previous findings [1]. Immunostaining with bcl-2 and bax antibodies revealed that both of these proteins were expressed in the cytoplasm of the cells in the interface membrane but they showed different cellular patterns. Bcl-2 was localized in a small number of lymphocyte-like cells while bax was expressed by large numbers of cells, mainly macrophages. The number of cells which expressed bcl-2 was significantly lower than that of bax (P<0.01). DNA fragment localization occurred mostly in a layer of cells (1- 3 cells deep) next to the implant surface. Again the level of DNA fragment-containing cells was significantly lower than that of bax positive cells (P<0.01). The results, for the first time, indicate that there is an apoptotic activity occurring in cells in the interface membrane, but not all the cells which express apoptosis-promoting protein (i.e. bax) will enter into the phase of cell death. PMID- 15347970 TI - Monocyte, macrophage and foreign body giant cell interactions with molecularly engineered surfaces. AB - To elucidate the mechanisms involved in monocyte/macrophage adhesion and fusion to form foreign body giant cells on molecularly engineered surfaces, we have utilized our in vitro culture system to examine surface chemistry effects, cytoskeletal reorganization and adhesive structure development, and cell receptor ligand interactions in in vitro foreign body giant cell formation. Utilizing silane-modified surfaces, monocyte/macrophage adhesion was essentially unaffected by surface chemistry, however the density of foreign body giant cells (FBGCs) was correlated with surface carbon content. An exception to the surface-independent macrophage adhesion were the alkyl-silane modified surfaces which exhibited reduced adhesion and FBGC formation. Utilizing confocal immunofluorescent techniques, cytoskeletal reorganization and adhesive structure development in in vitro FBGC formation was studied. Podosomes were identified as the adhesive structures in macrophages and FBGCs based on the presence of characteristic cytoplasmic proteins and F-actin at the ventral cell surface. Focal adhesion kinase (FAK) and focal adhesions were not identified as the adhesive structures in macrophages and FBGCs. In studying the effect of preadsorbed proteins on FBGC formation, fibronectin or vitronectin do not play major roles in initial monocyte/macrophage adhesion, whereas polystyrene surfaces modified with RGD exhibited significant FBGC formation. These studies identify the potential importance of surface chemistry-dependent conformational alterations which may occur in proteins adsorbed to surfaces and their potential involvement in receptor-ligand interactions. Significantly, preadsorption of alpha2 macroglobulin facilitated macrophage fusion and FBGC formation readily on the RGD surface in the absence of any additional serum proteins. As alpha2-macroglobulin receptors are not found on blood monocytes but are expressed only with macrophage development, these results point to a potential interaction between adsorbed 2 macroglobulin and its receptors on macrophages during macrophage development and fusion. These studies identify important surface independent and dependent effects in foreign body reaction development that may be important in the identification of biological design criteria for molecularly engineered surfaces and tissue engineered devices. PMID- 15347971 TI - Endothelial cell cultures as a tool in biomaterial research. AB - Progress in biocompatibility and tissue engineering would today be inconceivable without the aid of in vitro techniques. Endothelial cell cultures represent a valuable tool not just in haemocompatibility testing, but also in the concept of designing hybrid organs. In the past endothelial cells (EC) have frequently been used in cytotoxicity testing of materials, especially polymers, used in blood contacting implants, as well as for investigating seeding technologies for vascular prostheses. At present the exponential development both in theory and practice of cell and molecular biology of the endothelium offers great promise in the biomaterial field. Up until now this EC research field has mostly been non biomaterial orientated. Nevertheless, the relevance for biomaterial research is apparent. Four aspects will be concisely reviewed under the headings inflammation, with special reference to cell adhesion molecules (CAMs) and cytokines, angiogenesis, focusing on the healing response, signal transduction, presenting examples from cytokine- and metal ion-induced up-regulation of genes coding for CAMs, and, finally, endothelial functionality, with emphasis on the principal characteristics of the physiological endothelial phenotype. Finally, the application of these fields to three foci of biomaterial research will be discussed, emphasizing the role of EC culture techniques in controlling the host response to biomaterials (microvascular EC), controlling EC functionality (promoting positive effects and down-regulating negative effects), and tissue engineering (integration of EC into hybrid organs/biosensors). The need for more co-culture and three-dimensional models will be stressed and data from the authors' laboratory presented to illustrate these principles. PMID- 15347972 TI - Modulation of in vitro myogenesis induced by different polymer substrates. AB - The understanding of substrate dependence of cellular differentiation is important in the surface design of biocompatible artificial devices as well as cell-incorporated tissue engineered devices. In an attempt to understand some of the genetic and epigenetic aspects of the control of cell differentiation in the presence of two different materials, Chronoflex (CH) and plasma treated Chronoflex coated with Hyaluronan (CH-HA), we used primary cultures of human myogenic cells, a model that encompasses cell proliferation, migration, fusion, and differentiation dependent gene activation. By testing both the material samples on the growth of human myoblasts in primary cultures, we demonstrated that both CH and CH-HA substrates were able to support the cell growth since they did not affect cell count and DNA synthesis. On the contrary, the degree of myoblast differentiation, assessed as a function of creatine phosphokinase (CPK) activity on living cells, was completely different on the two biomaterials. Indeed, the amount of CPK increased on CH-HA cultured cells as a result of myotube formation, while CH grown myoblasts remained unfused and displayed no increase on the CPK activity even after 12 days culture. Moreover, the expression level of MyoD and myogenin mRNA, both related to myogenic cell differentiation, appeared extremely low in CH-grown cells, while they were rapidly induced in CH HA cultured myoblasts. PMID- 15347973 TI - Protein-mediated macrophage adhesion and activation on biomaterials: a model for modulating cell behavior. AB - The elucidation of proteins involved in biomaterial-modulated macrophage behavior is critical for the improvement of material performance and the initial exploration of material design capable of manipulating macrophage function for tissue engineering. In this paper, several in vitro and in vivo techniques are presented to demonstrate means of delineating a part of the complex molecular mechanisms involved in the interaction between biomaterial and macrophage adhesion and phenotypic development. The following conclusions were reached: (1) using radioimmunoassay, complement component C3 was found to be critical in mediating human macrophage adhesion on polyurethanes. (2) The presence of a diphenolic antioxidant additive in polyurethanes increased the propensity for complement upregulation but did not affect adherent macrophage density. (3) The subcutaneous cage-implant system was utilized to delineate interleukin-4 participation in the fusion of adherent macrophages to form foreign body giant cells in vivo in mice. The injection of purified interleukin-4 neutralizing antibody into the implanted cages significantly decreased the giant cell density; conversely, the giant cell density was significantly increased by the injection of recombinant interleukin-4 when compared with the controls. (4) The RGD and PHSRN amino acid sequences of the central cell binding domain and the PRRARV sequence of the C-terminal heparin binding domain of human plasma fibronectin were utilized to study the structure-functional relationship of protein in mediating macrophage behavior. Polyethyleneglycol-based networks grafted with the RGD-containing peptide supported higher adherent human macrophage density than surfaces grafted with other peptides. The formation of foreign body giant cell was highly dependent on the relative orientation between PHSRN and RGD domains located in a single peptide. PMID- 15347974 TI - In vitro and in vivo methods to determine the interactions of osteogenic cells with biomaterials. AB - To assess new biomaterials for possible use as bone graft substitutes, a number of techniques allow interactions with osteoblastic cells to be studied, with respect to effects on proliferation and differentiation of osteoprogenitors. In vitro models include the use of bone explant cultures, fetal rat calvarial derived osteoblast cells, primary stromal populations, transformed and non transformed cell lines and immortalized osteoblast cell lines. However, these assessments are limited by the extent of osteogenic differentiation and bone formation that can be observed in vitro, species differences and phenotypic drift of cells cultured in vitro. The use of in vivo experimental systems such as the segmental/calvarial bone defect model, the subcutaneous implant model and the diffusion chamber implantation model circumvent some of these issues and, in the appropriate model, provide data on efficacy, biocompatibility and osteointegration of a biomaterial. The combination of in vitro and in vivo approaches together with the development of new cell labeling techniques, in particular the ability to genetically mark and select specific human bone cell populations provides new avenues for their potential evaluation in combination with appropriate biomaterials for clinical use. These in vitro and in vivo techniques are reviewed and those recently developed for assessment of human osteogenic cells should be applicable to many other cell systems where knowledge of specific human tissue or cell interactions with biomaterials is required. PMID- 15347975 TI - Cytoskeletal rearrangement in K562 erythroleukaemic cells forced to grow on a positively charged polymer surface. AB - We have recently demonstrated that if human K562 erythroleukaemic cells, which normally grow in suspension, are grown on a positively-charged surface composed of polylysine, a transient reorganization of CD54 (ICAM-1), CD58 (LFA-3) and alphanubeta3 (vitronecin receptor), three important CAMs located on the cell membrane, takes place. In addition, changes of longer duration in membrane conductivity (ionic transport across the cell membrane) and membrane permittivity (static distribution of charges across the cell membrane), indicating more permanent structural as well as functional alterations in the cell membrane, were also observed [2]. Because of the close interrelationship which exists between the cell membrane, CAMs and the cytoskeleton, changes in this intracellular network as well as in the surface morphology of K562 cells grown on the positively-charged polymer, polylysine, were examined. In particular, actin and tubulin were investigated qualitatively and quantitatively by immunofluorescence microscopy and flow cytometry, respectively, while the cell surface was studied by scanning electron microscopy (SEM). The data indicate that when K562 cells are grown onto polylysine no quantitative changes occurred to the cytoskeletal elements even if these were rearranged and that the cell membrane surface is also greatly altered. These results are discussed in light of the pivotal role played by CAMs and the cell cytoskeleton in transducing environmental stimuli, in this case those provided by a positive charge, from the cell membrane to the inside of the cell. PMID- 15347976 TI - The development of in vitro biocompatibility tests for the evaluation of intraocular biomaterials. AB - Recent developments in ocular implant technology require the in vitro evaluation of ocular compatibility in early stage development programs. This requires an understanding and appreciation of the biological interactions which occur in the ocular environment and their relevance with respect to the clinical complications associated with surgical implantation of devices. This paper describes the development of a series of clinically reflective in vitro assays for assessing the potential ocular compatibility of novel intraocular lens materials. Staphylococcus epidermidis attachment, fibrinogen adsorption, mouse embryo fibroblast 3T3 adhesion and proliferation, primary rabbit lens cell adhesion, human peripheral blood macrophage adhesion and granulocyte activation tests were employed to evaluate two widely used intraocular biomaterials poly(methyl methacrylate) (PMMA) and silicone, and a novel biomimetic phosphorylcholine-based coating (PC). The performance of these materials in the in vitro assays was compared to their ability to reduce postoperative inflammation in vivo in a rabbit model. The results demonstrated that the in vitro assays described here are predictive of in vivo ocular compatibility. These assays offer a more relevant means of assessing the ocular compatibility of biomaterials than those presently required by the authorities for regulatory approval of medical devices and implants. PMID- 15347977 TI - MMA/MPEOMA copolymers as coating materials for improved blood compatibility: protein adsorption study. AB - Surface-induced thrombosis remains one of the main problems in the development of blood-contacting devices. When a foreign surface comes in contact with blood, the initial blood response is adsorption of blood proteins, followed by platelet adhesion and activation, leading to thrombus formation. A particularly effective polymer for the prevention of protein adsorption and platelet adhesion appears to be polyethylene oxide (PEO). In this study, water-insoluble copolymers of methyl methacrylate (MMA) and methoxy PEO monomethacrylates (MPEOMA) with different PEO molecular weights (200, 400, and 1000) and monomer composition were synthesized and characterized by gel permeation chromatography and 1H-nuclear magnetic resonance spectroscopy. The synthesized copolymers were coated on glass slides by a spin coating method to prepare PEO-rich surfaces as blood-compatible surfaces. The surface properties of the copolymers and their interaction with blood proteins (albumin, gamma-globulin, fibrinogen, and plasma proteins) were investigated by the measurement of water contact angles and by electron spectroscopy for chemical analysis, respectively. It was observed that the protein adsorption on the copolymer surfaces decreased with increasing PEO molecular weight and MPEOMA content in the copolymers. The copolymers with long PEO chains in MPEOMA (MMA/MPEO1000MA copolymers) were effective in preventing protein adsorption, even though their MPEOMA content was less than the copolymers with shorter PEO chains. PMID- 15347978 TI - Linear poly(ethylene oxide)-based polyurethane hydrogels: polyurethane-ureas and polyurethane-amides. AB - Over the last 30 years, water-swellable and water-insoluble hydrogels have been extensively investigated and developed, leading to a large family of materials which have found uses in a wide range of biomedical applications. While hydrogels usually present a crosslinked structure, linear polyurethane-ureas (PUUs) based on poly(ethylene oxide) have been shown to be able to absorb and swell with aqueous media without dissolving. This behavior is due to the phase separated domain morphology, where hydrogen bonded urethane/urea hard segment domains are dispersed in a PEO soft segment domain. This work investigates the possibility of obtaining linear poly(ethylene oxide)-based polyurethane-amide (PUA) hydrogels using two amide diols as chain extenders, a mono amide diol (AD) and a diamide diol (DD), and a dicarboxylic acid (maleic acid, MA). Poly(ethylene oxide) based PUAs were obtained using a "one-shot" bulk polymerization technique. The chemicophysical characterization and water-solubility tests showed that these materials, while having molecular weights similar to the PUUs, do not possess sufficient phase separation, hydrogen bonding and hydrophobicity of the hard segment domains to exhibit hydrogel behavior. Crosslinked PUAs using maleic acid as chain extender show interesting hydrogel properties. PMID- 15347979 TI - Resorbable polyacrylic hydrogels derived from vitamin E and their application in the healing of tendons. AB - A hydrogel containing vitamin E (alpha-tocopherol) was prepared by free radical polymerization of 2-hydroxyethyl methacrylate (HEMA) and alpha-tocopheryl methacrylate (VEMA), the latter being synthesized previously to its use. The hydrogel containing 20 wt % of VEMA showed equilibrium water content in the range of those of hydrogel networks, at any pH. The swelling of the hydrogel followed Fick's law, indicating that sorption of water molecules is controlled by diffusion, although the values of diffusion coefficients for the VEMA-containing hydrogel were lower than those of poly-HEMA in any medium. Surface characterization of the VEMA-containing hydrogel revealed a decrease in the surface energy of solid owing to a decrease of the polar component mainly. The application of finely powdered xerogel of HEMA-VEMA copolymer bearing 20 wt % of the vitamin E derivative gave a very fast and positive response showing an activated regeneration capacity, probably due to the stimulation of the cellular proliferation or the more plausible effect, the cellular protection associated to the antioxidant properties of the vitamin E residue. PMID- 15347980 TI - Investigation into the mechanism of bacterial adhesion to hydrogel-coated surfaces. AB - As a model for hydrogel-coated biomaterials, self-assembled monolayers of polyoxyethylene (POE) derivatives on sheets of polymeric biomaterials were prepared. The POE derivatives consisted of hydrophilic chains with different lengths and a long-chain alkyl group that served as an anchor function. The coatings obtained were analyzed with XPS and contact angle measurements showing hydrophilic chains of different lengths extending away from the surface. Bacterial adhesion was measured with a clinically relevant Klebsiella pneumoniae type strain and measurements reproduced 12 times. Bacterial adhesion decreased markedly with increasing hydrophilic chain length. Based upon these findings a new model for bacterial adhesion to hydrogel-coated surfaces is suggested: steric repulsion effects that increase with increasing chain length of grafted hydrophilic chains play an important role in bacterial adhesion to hydrogel coated surfaces. PMID- 15347981 TI - Biodegradable drug delivery system for the treatment of bone infection and repair. AB - A drug delivery system (DDS) which provides a sustained release of antibiotics at the focal site either singly, or in combination with a bone stimulating factor could both eliminate infection and increase the number of potentially healthy osteogenic cells. In this study, we address the use of a degradable gelatin DDS, for the combined release of therapeutic levels of both gentamicin and growth hormone (GH). An initial bolus release was observed during the first 24 h followed by a reduced, but sustained, release for both drugs up to 14 days. Bioactivity of gentamicin was demonstrated by growth inhibition of Staphylococcus aureus for over 96 h with a mean zone of inhibition of 29.4 mm (+/-0.19) for the time period studied. Furthermore, GH was shown to have a direct effect on primary human osteoblast-like (HOB) cells, stimulating proliferation and enhancing their differentiation. Site-specific drug delivery offers the advantage of localizing a drug directly at the target site, thus minimizing systemic effects. The results of this study suggest that gelatin is a good DDS for the combined release of drugs. In addition, gelatin is both biocompatible and biodegradable, thus making it a promising DDS for the management of acute and chronic bone and tissue infection such as osteomyelitis. PMID- 15347982 TI - Fibers by bioresorbable poly(ester-ether-ester)s as potential suture threads: a preliminary investigation. AB - Fibers made by a bioresorbable poly(epsilon-caprolactone)-block-poly(oxyethylene) block-poly(epsilon-caprolactone) copolymer, having a number average molecular mass of about 200,000 Da and an average molar composition of 66% oxycaproyl units and 34% oxyethylene units, were melt-spun, with the aim at using them as suture threads. Their properties were investigated by the stress-strain test and by differential scanning calorimetry (DSC). The results obtained show that the properties of this material depend very strongly on the alignment of its macromolecules. In particular, the only partial alignment, obtainable by a relatively moderate drawing just after the extrusion, leads to values of elongation at break too high for use of the fibers as suture threads. The DSC analysis reveals interesting properties of the material, but also confirms their strong dependence on the extrusion procedure and on the mechanical treatment. In conclusion, the results of this preliminary study show that the spinning technique must be improved, and that further investigations are necessary to ascertain the possibility of using these poly(ester-ether-ester)s for the fabrication of suture threads. PMID- 15347983 TI - Quantification of bone ingrowth within bone-derived porous hydroxyapatite implants of varying density. AB - Hydroxyapatite has been investigated for use in the osseous environment for over 20 years and the biocompatibility of the ceramic and its osseoconductive behavior is well established. Therefore, the use of porous hydroxyapatite for the repair of osseous defects seems promising with potential for complete penetration of osseous tissue and restoration of vascularity throughout the repair site. However, there have been few systematic studies of the effects of physical properties such as macropore size and pore connectivity on the rate and quality of bone integration within porous hydroxyapatite implants. This paper quantifies the early biological response to a well-characterized series of implants with uniform microstructure and phase composition, but differing macrostructures and demonstrates the dependence of the rate of osseointegration on the apparent density of porous hydroxyapatite as a function of pore connectivity. Furthermore, compression testing established that bony ingrowth has a strong reinforcing effect on porous hydroxyapatite implants, which is more pronounced in the lower density implants, as a result of a greater relative volume of bone ingrowth. PMID- 15347984 TI - Macroporous hydroxyapatite as alloplastic material for dental applications. AB - In the dentistry field, synthetic hydroxyapatite can be conveniently used as semiabsorbable alloplastic material to solve a number of clinical problems. The hydroxyapatite structure influences bone ingrowth as well as its resorption. In particular, pore size must exceed 100 microm to allow new bone ingrowth. Because of its brittleness, highly porous hydroxyapatite is difficult to handle without causing damage, and, therefore, its mechanical performance has to be improved placing the material on a dense hydroxyapatite substrate. Dense/porous hydroxyapatite laminates can be obtained by slip casting technology. The method starts with the preparation of a ceramic powder-binder-solvent system. This slurry produces, after solvent evaporation, a soft green tape, that is successively baked at 300 degrees C to remove all organic component, and finally sintered at high temperature (1200 degrees C). The material porosity cannot be significantly modified by changes in the slurry composition and sintering temperature; therefore, the macroporous hydroxyapatite layer must be obtained using a porous hydroxyapatite powder. The powder is prepared by grinding of green tape pieces and the resulting coarse material is successively baked at 300 degrees C. Such material is mixed with a polymer solution and cast on a green tape substrate. Layer fractures are not observed in both film bulks and interface, because during sintering the consolidation of two layers happens simultaneously. In the resulting material, the first layer consisted of macroporous hydroxyapatite with high osteoconductive properties, and the second layer was a dense hydroxyapatite substrate able to improve the laminate mechanical properties. PMID- 15347985 TI - Periodontal membranes from composites of hydroxyapatite and bioresorbable block copolymers. AB - Biomembranes are frequently proposed as devices for "guided bone regeneration." Such membranes consist generally of a thin sheet of polymeric material, mostly textured from polymeric yarns or clots, which all have a diffuse very fine winding porosity. The cross-section size of the holes of such porosity is nanometric (diameter < 0.1 microm); thus these holes can be indicated as nanoholes. Whatever the method of production, the surface density of nanoholes (number per square centimeter) has to be as high as possible. It is important also that no variation of this density occurs. The fine dimension of these microholes allows the crossing of small molecules (O2, CO2, H2O, sugars, many nutritional organic compounds and even some simple proteins) but not other larger molecules and particulates, including cells of any kind. These biomembranes have, consequently, a semipermeable behavior, providing the functional role which is the interposition of a barrier for the cells, separating the bone from the surrounding soft tissues. The kinetic of proliferation of osteoblasts is lower than that of fibroblasts. Most membranes of this kind are not resorbable. The main problem for the resorbable ones is the speed of size increase of the holes during the time. Their diameter must not exceed a threshold value until the reconstruction of bone is complete, otherwise soft tissue cells will invade the growing bone tissue with formation of undesirable mixed tissue. The present paper deals with a resorbable membrane made with a composite polymer/ceramic. A poly(epsilon-caprolactone)-block-poly(oxyethylene)-block-poly(epsilon caprolactone) copolymer is the polymeric matrix which contains dispersed ceramic hydroxyapatite microgranules, a stiff filling additive. The main possible use is that of periodontal membranes. The copolymer, obtained by thermal polymerization of epsilon-caprolactone onto poly(ethylene-glycol), presents good biological tolerance, is resorbable under physiological conditions and can promote cell growth. Histological tests, performed 6 months after implantation, showed that the polymeric matrix is almost totally resorbed. New-formed bone colonizes even the innermost parts of the membrane, with bone trabeculae closely surrounding the hydroxyapatite granules. PMID- 15347986 TI - In vitro reconstructed tissues on hyaluronan-based temporary scaffolding. AB - Tissue engineering offers the possibility to reconstruct tissue substitutes in order to replace lost or damaged tissues. The availability of appropriate biomaterial devices is essential to allow in vitro cultured cells to behave as in the original tissues in vivo. In our studies we utilized a seminatural biomaterial made up by the benzyl ester of hyaluronan to grow keratinocytes, fibroblasts and chondrocytes. Keratinocytes and fibroblasts were isolated from human foreskin. Cells were separately cultured on two different hyaluronan based biomaterial devices for the first 15 days and then co-cultured for an additional period of 2 weeks. Keratinocytes gave rise to a well-differentiated epithelial layer, while fibroblasts were able to synthesize all the main extracellular molecules inside the biomaterial spaces, forming dermal-like tissues. When these two tissues were co-cultured, a skin equivalent was formed with a dermal epidermal junction. Chondrocytes were obtained from chick-embryo sterna and cultured for 21 days inside a non-woven scaffolding made up of a hyaluronan-based biomaterial. Cells were able to organize themselves into nodules embedded in a dense metachromatic substance in which type II collagen was present. Data from this study suggest that this novel class of hyaluronan derived biomaterials is suitable for different cell culture and in vitro tissue reconstruction. PMID- 15347987 TI - Behavior of tissue-engineered human cartilage after transplantation into nude mice. AB - Cartilage lacks the ability to regenerate structural defects. Therefore, autologous grafting has been used routinely to replace cartilaginous lesions. Because tissue engineering of human cartilage with the help of bioresorbable polymer scaffolds is possible in experimental models, the demand for the clinical application grows. In this study we present an analysis of the behavior of transplants made of human chondrocyte pools, agarose and the resorbable polymer scaffold Ethisorb and a preliminary comparison with transplants made of single patients' cells and Ethisorb but without the additional ingredient agarose. Chondrocytes were isolated from the matrix of human septal cartilage by enzymatic digestion. The pool cells were kept in monolayer culture for 2 weeks, the single patients' cells for 3-4 weeks. Chondrocyte pools were suspended in agarose and seeded into the resorbable polymer scaffold Ethisorb. Single patients' cells were seeded without agarose. All cell-polymer constructs were kept in perfusion culture for 10-14 days and transplanted subcutaneously into thymusaplastic nude mice. Additionally we implanted Ethisorb fleeces embedded in agarose without chondrocytes. After 6, 12 and 24 weeks the animals were sacrificed and the specimens were explanted and analyzed histochemically and immunohistochemically. Polymer scaffolds not seeded with chondrocytes did not show cartilage formation. Resorption was complete after 12 weeks in vivo. Transplants from cell pools remained mechanically stable over 24 weeks apart from four transplants that were resorbed completely. Cartilage formation was observed in all pool-specimens with the presence of chondronic structures and a homogeneous matrix containing hyaline cartilage-specific matrix molecules such as collagen type II. Single patients' transplants showed hyaline cartilage matrix synthesis and mechanical stability as well. Chondrocyte pools are a suitable method to study cartilage engineering of human cells in vitro and in vivo in experimental models. Under clinical conditions it is, however, necessary to study the generation of cartilage from single patients' cells. We showed that it is possible without additional ingredients such as agarose. However, variations in the preliminary results show that the clinical application with human cells is more difficult than one would expect when using human chondrocyte pools. Further studies need to be performed to find out which individual factors influence the in vitro engineered cartilage's fate in vivo. PMID- 15347988 TI - Material properties of absorbable self-reinforced fibrillated poly-96L/4 D lactide (SR-PLA96) rods; a study in vitro and in vivo. AB - A study was carried out to investigate changes in mechanical properties and degradation of self-reinforced fibrillated poly-96L/4D-lactide (SR-PLA96) rods in vitro and in vivo. The viscosity-average molecular weight, Mv, of the intact sterile (gamma irradiated) rods was around 50,000 g mol(-1). The SR-PLA96 rods of diameter 1.1 mm by 30 mm and diameter 4.5 mm by 50 mm were immersed in phosphate buffered saline or implanted in the dorsal subcutis of rabbits. Bending, shear and torsion strength and bending modulus, together with the changes of viscosity and crystallinity, were measured up to 24 wk. The strength values showed only a slight decrease during the follow-up period with the exception of torsion strength, that decreased to 52% of the initial value during 24 wk. There were no statistically significant differences in the strength retention between in vitro and in vivo groups. Crystallinity increased over time, being 46-49% at 24 wk. The Mv of the rods decreased over 50% by 24 wk. These promising results motivated us to continue the studies with the fixation of experimental cortical bone osteotomies with SR-PLA96 intramedullary rods. PMID- 15347989 TI - Mechanical properties of sintered hydroxyapatite and tricalcium phosphate ceramic. AB - The ultimate values for compressive strength, Young's modulus, and toughness of cylindrical specimens of unitary aspect ratios and uniform grain-size distributions were extrapolated for hydroxyapatite (HAP) to 70 MPa, 9.2 GPa, and 0.36 J cm(-3), and for tricalcium phosphate (TCP), to 315 MPa, 21 GPa, and 2.34 J cm(-3). For total volume porosities of 50%, the corresponding values were determined: for HAP, 9.3 MPa, 1.2 GPa, 0.042 J cm(-3), for TCP, 13 MPa, 1.6 GP, 0.077 J cm(-3). Porosities of HAP specimens ranged from 3%-50%; TCP from 10%-70%. Two pore-size distributions were employed. Exponential dependencies of the mechanical properties were found upon porosity (p < 0.0001). No differences in measured mechanical properties, as determined in compression, could be attributed to pore size. The superiority of TCP increases with density and suggests that a larger or more selective pore-size distribution could be effectively employed in TCP biological implants. This work also suggests the dominant role of secondary calcium phosphates in increasing compressive strengths. PMID- 15347990 TI - Human endothelial cell attachment to and growth on polypyrrole-heparin is vitronectin dependent. AB - Composite materials comprised of the electrically conducting polymer, polypyrrole, with a variety of biologically active molecules, e.g. proteins or polysaccharides, are emerging as a novel class of "smart" biomaterials. In the present work we have studied the utility of a heparin-polypyrrole composite as a substrate for human umbilical vein endothelial cell (HUVEC) growth. We found that the polymer composites were well suited to support cell attachment and growth; displaying low surface hydrophobicity (water contact angle of approximately 20 degrees ) and roughness, Rq of approximately 10-12 nm. Doubling times for HUVEC on heparin-polypyrrole were greater than observed for gelatin-coated tissue culture polystyrene (44 and 36 h, respectively), however, the cells did proliferate to cover the polymer in an even monolayer. The initial mechanism of attachment and subsequent proliferation of HUVEC on heparin-polypyrrole was critically dependent on the presence of the serum adhesion glycoprotein vitronectin. Polymers that were composed of polypyrrole and sodium nitrate were more hydrophobic than heparin-polypyrrole and they did not support HUVEC growth. Given the relative ease with which these polymer composites can be electrochemically synthesized, the diverse range of cellular "signal agents", e.g. growth factors, that can be incorporated within them, and the high degree of control that can be achieved in the release-surface exposure of these agents, we suggest that polypyrrole composites could serve a useful role as "smart" biomaterials in the near future. PMID- 15347991 TI - Synthesis and properties of poly(lactic acid). AB - Poly(D,L lactic acid) was prepared by bulk polymerization of D,L lactide, both under atmospheric pressure and in vacuum. The obtained polymeric products were characterized in terms of molecular weight, Mw, melting point, calorimetric response and swelling behaviour. All products were amorphous. Their molecular weights were determined by viscosimetry and ranged from 2x10(3) to 9x10(4). Similarly, the melting points ranged from 90 to 210 degrees C. Swelling experiments, with specimens immersed in buffer solutions, showed that hydrolytic degradation started in a few days for the low Mw material, whereas for the higher molecular weight products it took much longer and probably followed a two-stage mechanism. This study suggests that the high molecular weight material could be an interesting carrier for the preparation of controlled release products, in cases where prolonged delivery is necessary. PMID- 15347992 TI - Surface characterization of implant materials c.p. Ti, Ti-6Al-7Nb and Ti-6Al-4V with different pretreatments. AB - The biocompatibility of commercially pure titanium and its alloys is closely related to their surface properties, with both the composition of the protecting oxide film and the surface topography playing an important role. Surfaces of commercially pure titanium and of the two alloys Ti-6Al-7Nb and Ti-6Al-4V (wt %) have been investigated following three different pretreatments: polishing, nitric acid passivation and pickling in nitric acid-hydrogen fluoride. Nitric acid treatment is found to substantially reduce the concentration of surface contaminants present after polishing. The natural 4-6 nm thick oxide layer on commercially pure titanium is composed of titanium oxide in different oxidation states (TiO2, Ti2O3 and TiO), while for the alloys, aluminium and niobium or vanadium are additionally present in oxidized form (Al2O3, Nb2O5 or V-oxides). The concentrations of the alloying elements at the surface are shown to be strongly dependent on the pretreatment process. While pickling increases the surface roughness of both commercially pure titanium and the alloys, different mechanisms appear to be involved. In the case of commercially pure titanium, the dissolution rate depends on grain orientation, whereas in the case of the two alloys, selective alpha-phase dissolution and enrichment of the beta-phase appears to occur. PMID- 15347993 TI - Characterization and histological analyses of a coral-collagen composite used for bone-replacement graft material: a report of clinical cases. AB - Several studies, devoted to the osteogenic potentialities of natural CaCO3 have already been reported. However, it seems questionable if the data obtained from natural calcium carbonates can be extrapolated to a composite biomaterial incorporating coralline material. For these reasons, in the present investigations the structural and crystallographic features of the biomaterial (Biocoral gel) were thoroughly analyzed prior to implantation, with the aid of X ray diffraction and electron microscopy. Then, biopsied samples, taken from Biocoral gel-filled sites, respectively after 7, 8, 9, 12 and 29 mon implantation, were studied with optical and electron microscopy. It could be concluded from the histological analyses of the biopsies, that mineral still remained after long implantation periods. This composite biomaterial may thus be considered for uses in clinical situations where neither incorporation nor dissolution of the implanted biomaterial are essential, i.e. maintenance of edentulous ridge volume. PMID- 15347994 TI - Biodegradable tube implants in experimental glaucoma surgery in the rabbit. AB - Although ocular drainage implants are manufactured from biocombatible materials to reduce foreign-body reaction, the formation of excessive scar tissue around the implant is a common cause for implant failure. In this study, the suitability of poly(D, L-lactide-co-glycolide) copolymer, impregnated with an antiproliferative agent retinoic acid, was evaluated as a material for biodegradable tubular implants, as well as the duration and magnitude of the intraocular pressure reduction obtained with the prototype implant. Subconjunctivally placed retinoid-impregnated polymer particles caused a milder inflammatory reaction than plain polymer, and the layer of connective tissue around the material was thinner after the follow-up period of 60 d. In the anterior chamber, the inflammatory response elicited by the material was milder than subconjunctivally. The plain polymer caused a transiently stronger reaction than the retinoid-impregnated polymer, but after 60 d no difference was evident between the two materials. In all operated eyes with the tubular implant, the intraocular pressure was statistically significantly lower (p<0.05) than in control eyes for 9 wk after the operation. The intraocular pressure of the eyes with the retinoid-impregnated implant was statistically significantly lower (p<0.05) than in eyes with a plain polymer implant for up to 7 wk post operatively. However, the use of retinoid did not prolong the effective functioning time of the implants. PMID- 15347995 TI - Effects of water-soluble component content on cephalexin release from bioactive bone cement consisting of bis-GMA/TEGDMA resin and bioactive glass ceramics. AB - The effect of the amount of a water-soluble, lactose, on cephalexin (CEX) release from bioactive bone cement consisting of bisphenol-alpha-glycidyl methacrylate (bis-GMA), triethylene-glycol dimethacrylate (TEGDMA) resin and apatite- and wollastonite-containing glass-ceramic (A-W GC) powder was investigated. A-W GC powder containing 5% CEX and lactose powders hardened within 5 min after mixing with bis-GMA/TEGDMA resin, and furthermore its compressive strength was expected to be higher than that of polymethylmethacrylate cement. In vitro CEX release from bioactive bone cement pellets in a simulated body fluid at pH 7.25 and 37 degrees C continued for more than 2 wk. The drug-release rate increased with increasing amount of lactose powder in the mixture. CEX release profiles followed the Higuchi equation in the initial stage, but not in later stages. As hydroxyapatite was precipitated out on the cement surface, the CEX release rate decreased. The micropore distribution of the cements measured by mercury porosimetry also supported the variation in drug release due to cement porosity being mainly a result of the dissolution of lactose in the cements. These results suggest that the rate of CEX release from bioactive bone cement could be controlled by varying the amount of lactose in the cement system. PMID- 15347996 TI - Estrogenic activity of chemicals for dental and similar use in vitro. AB - The estrogenic activities of chemicals for dental and similar use were tested by a reporter gene assay (yeast two-hybrid system) and an estrogen/estrogen receptor (ER-alpha) competition binding assay (fluorescence polarization system). Among the 10 chemicals [bisphenol-A (BPA), bis-2-hydroxypropyl methacrylate (Bis-GMA), triethylene glycol dimethacrylate (TEGDMA), methyl methacrylate (MMA) and 2 hydroxyethyl methacrylate (HEMA), dibutyl phthalate (DBP), n-butyl benzyl phthalate (BBP), n-butyl phthalyl n-butyl glycolate (BPBG), di-2-ethylhexyl phthalate (DEHP), and di-2-ethylhexyl adipate (DOA)], which were diluted with DMSO to concentrations ranging from 5 x 10(-7) to 5 x 10(-3) M, and 17beta estradiol (E2) as a positive control, BPA and BBP showed estrogenic activity in these two assays, while the remaining eight chemicals did not at the concentrations tested. Additional data, together with in vivo and epidemiological examinations, are required. Such investigations should also provide information on the validity of these methods for testing the estrogenic activity of chemicals. PMID- 15347997 TI - A study of alternative metal particle structures and mixtures for dental amalgams based on mercury additions. AB - The perception that mercury in dental amalgam is toxic to the human organism has prompted worldwide efforts by the scientific community to develop alternative amalgam-like materials that utilize little or no mercury. In this investigation, an attempt is made to develop a new dental alloy system by adding liquid mercury to silver-coated Ag4Sn intermetallic particles in lesser amounts than are used in conventional amalgam alloys. An effort to precipitate the important eta-prime (Cu6Sn5) phase was made by adding pure Cu and Sn powders to the alloy formulation during trituration. Tytin a popular Ag-Sn-Cu single-composition, spray-atomized conventional dental alloy was used as the control to obtain baseline data for comparisons of microstructures and mechanical properties. Amalgamation of the coated particles with mercury, with or without the addition of Cu and Sn powders, mostly produced specimens with chemically non-coherent microstructures that were relatively weak in compression. These results were due, in part, to mercury's inability to chemically wet the Ag-coated particles and Cu and Sn powders because of naturally occurring surface oxide films. The strongest specimens tested had silver dendritic coatings, resulting in compression strength values up to 40% of the control's. Their higher strength is attributed to mechanical interlocking at the particle/matrix interfaces. PMID- 15347998 TI - Thermal analysis studies of poly(etheretherketone)/hydroxyapatite biocomposite mixtures. AB - Biocomposite formulations which have the potential to combine the proven mechanical performance of poly(etheretherketone) (PEEK) with the inherent bioactivity of hydroxyapatite (HA), may have a utility as load-bearing materials in a medical implant context. The effect of thermal processing on the relevant properties of the PEEK and/or HA components in any fabricated composite structure is, however, an important consideration for their effective exploitation. This paper reports the results of a detailed thermal characterization study of a series of PEEK/HA mixtures using thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and modulated differential scanning calorimetry (MDSC). The TGA analyses show minimal weight loss for all of the mixtures and for a pure PEEK sample up to approximately 530 degrees C. Above this point there is a sharp on-set of decomposition for the PEEK component in each case. The temperature at which this feature occurs varies for each mixture in the approximate range 539-556 degrees C. This observation is supported by the presence of exotherms in the corresponding DSC scans, in the same temperature region, which are also assigned to PEEK decomposition. The temperature at which the degradation on-set occurs is found to decrease with increasing HA contribution. The use of the modulated DSC technique allows a number of important thermal events, not easily identifiable from the data obtained by the conventional method, to be clearly observed. In particular, the glass transition temperature (Tg) of the polymer can now be accurately determined. Using these thermal analysis data, calculations of the % crystallinity of PEEK in the mixtures have been made and compared with that of a 100% polymer sample. From these studies it is evident that the presence of HA does not adversely affect the degree of crystallinity of the PEEK component in the mixtures of interest over the thermal range studied. PMID- 15347999 TI - Mechanical properties of hydroxyapatite reinforced poly(ethylmethacrylate) bone cement after immersion in a physiological solution: influence of a silane coupling agent. AB - PEMA-based bone cement has previously been shown to possess many advantages over traditional PMMA cements. One of these is the option of adding up to 40 wt % HA without a decrease in static mechanical strength, thus providing the potential for enhanced bioactivity. Bone cement, in vivo, is subjected to an aqueous environment and therefore, it is important to understand the influence of this upon the mechanical integrity of experimental cements. In this current investigation the static and dynamic properties of PEMA cement, with and without 30 wt % untreated and silanated HA, were examined after periods of immersion in Ringer's solution. A commercial PMMA cement was also tested in a similar manner. Relatively small changes in static mechanical properties were observed after 12 weeks storage for the PEMA cements, the largest change being for the PEMA cement reinforced with silanated HA. The PMMA cement exhibited the largest change in static strength with a decrease of 16.6%. In contrast to these results, the fatigue properties of the PEMA cements were found to decrease significantly after storage in Ringer's solution, again with the largest changes to the PEMA cement reinforced with silanated HA. This effect was attributed to the reduction in efficiency of the silane coupling agent in the presence of water. The fatigue resistance of the PMMA cement was not reduced after immersion in a saline environment. PMID- 15348000 TI - A percutaneous device to study glucose kinetics in subcutaneous tissue fluid. AB - In the current study subcutaneous glucose kinetics were investigated in tissue fluid collected with a percutaneous device (PD). PDs containing a subcutaneous tissue chamber were implanted subcutaneously in New Zealand white rabbits. Sintered titanium fiber mesh sheets were used for subcutaneous anchorage of the PD. The bottom of the subcutaneous tissue chamber was either covered with a titanium fiber mesh sheet, a cellulose acetate membrane, or left uncovered. Subcutaneous glucose kinetics were determined after injection of octreotide and glucagon. The tissue reaction to the implants was evaluated histologically. No dynamic relationship was observed between glycaemia and subcutaneous tissue fluid glucose for all membrane covered devices. Histological evaluation showed that the presence of a seroma cavity in combination with obstruction of the membrane prevented adjustment of the subcutaneous glucose concentration in response to changes in glycaemia. In the uncovered devices, on the other hand, changes in glycaemia were reflected in subcutaneous tissue fluid. Our results prove that it is possible to measure changes in the glucose concentration in subcutaneous tissue fluid collected with a percutaneous device. Therefore, we conclude that a percutaneous device has an application as model to study the in vivo performance of implantable glucose sensors. The use of porous membranes in such devices has to be avoided. PMID- 15348001 TI - Synthesis of acrylate functional telechelic poly(lactic acid) oligomer by transesterification. AB - The controlled synthesis of low molecular weight (Mn from 700 to 10 000 g/mol) poly(lactic acid) (PLA) telechelic oligomers end-capped with acrylate groups by a one-step reaction was investigated. A transesterification reaction was carried out in solution with a Lewis acid titanium catalyst using a high molecular weight PLA and a low molar mass diacrylate. End-functionalization was demonstrated by proton NMR spectroscopy which was also used for quantitative analysis and number average molecular weight determination using the ratio between the acrylic chain ends to the main groups of poly(D,L-lactic acid). The formation of low molar mass oligomers from high molecular weight poly(lactic acid) was verified by gel permeation chromatography. Matrix-assisted laser desorption ionization time-of flight mass spectrometry showed that the above oligomerization was accompanied by the formation of cyclic compounds. By these means the feasability of a simple transesterification for a controlled synthesis of telechelic oligomers with molecular mass being a function of the added amount of diacrylate has been demonstrated. The glass transition temperatures of the elaborated oligomers varied from -35 to -5 degrees C. Subsequent thermal crosslinking was performed using benzoyl peroxide which enabled the formation of amorphous networks with Tg's close to the body temperature of 40 degrees C. Upon storage in a humid atmosphere the initially fairly hard and brittle networks became, due to hydrolysis, progressively more flexible thus demonstrating the potential biodegradability of these materials. PMID- 15348002 TI - Effect of iron state on crystallization and dissolution in Fe2O3-CaO-SiO2 glasses. AB - The possibility of iron-containing glasses as thermoseeds for hyperthermia of bone tumor was reported previously. There is, however, no report about the effect of iron state on the crystallization of magnetite and the resultant properties. The iron states were determined by Mossbauer spectroscopy in Fe2O3-CaO-SiO2 system. It was found that the higher CaO content interrupts the crystallization of magnetite crystallites as well as the oxidation of iron, that is, the transformation from Fe3+ to Fe2+. A sample containing large amounts of Fe2+ showed the faster increment of temperature when the alternating magnetic field was applied. In order to use the thermoseed for a hyperthermia, we can say that the composition with low CaO content is most useful. PMID- 15348003 TI - An improved peel test method for measurement of adhesion to biomaterials. AB - Adhesion of tissues to biomaterials is desirable to prevent bacterial proliferation and for epithelial/transmucosal sealing of transcutaneous appliances, but can be counter-productive elsewhere, e.g. implants contacting tendons or maxillofacial subcutaneous tissue. It is therefore important to gauge adhesion strength of tissues to biomaterials before clinical use. Peel-testing is widely used for industrial product adhesion monitoring, but has rarely been applied biomedically. Here we describe peel-testing instrumentation designed for testing adherence of soft tissues to biomaterials. It offers the advantage that a 90 degrees angle between peel and substrate is maintained, simplifying determination of applied normal forces separating tissue layers from material surfaces. The device is portable and can be brought directly to the specimen removal site. This minimizes time delays between explantation and testing, maintaining the tissue/biomaterial interface in the freshest possible state closely approximating in vivo conditions, and so avoids measurement artifacts. So far, the instrument has been used to test adhesion of tape to a biomaterial surface (for determining the device's technical performance), assess strength of tissue adhesives, and measure adhesion of subcutaneous tissue to orthopaedic biomaterials. However, its versatility suggests additional applications for the peel-tester where adhesion of soft tissue to biomaterials is of interest. PMID- 15348004 TI - Morphological study of hydroxyapatite nanocrystal suspension. AB - Nanometer size hydroxyapatite (HA) crystals are prepared by a wet chemical precipitation method at different synthesis temperatures and with various reactant addition rates. The resulting aqueous suspensions are studied in terms of morphology (transmission electron microscope, specific surface area), phase (X ray diffraction, electron diffraction and infrared spectroscopy) and rheological properties. This work shows that shape, size and specific surface area of the HA nanoparticles are very sensitive to the reaction temperature and also to the reactant addition rate. The measured pH at the end of synthesis, which is strongly linked with the reactant addition rate, is a key parameter which can be used to determine the purity of the synthesized HA nanocrystal and also for the stabilization (dispersion) of the suspension. HA nanoparticles synthesized at low temperature (T < 60 degrees C) are monocrystalline. A transition temperature (T=60 degrees C) can be defined as a limit for the synthesis of monocrystalline HA nanocrystals, above this critical temperature nanocrystals become polycrystalline. HA monocrystals adopt a needle shape and are oriented following the c-axis of the hexagonal HA structure. The as-synthesized suspension is then concentrated and the effect of a dispersing agent addition, which is needed to get a high solid/liquid ratio coupled with good flowability of the suspension, is also shown, because this suspension is used in the suspension plasma spraying process. PMID- 15348005 TI - Mechanical properties of dental investment materials. AB - Measurement of the elastic modulus (E) of investment materials has been difficult because of their low strength. However, these values are essential for engineering simulation and there are many methods available to assess the elasticity of materials. The present study compared two different methods with one of the methods being non-destructive in nature and can be used for specimens prepared for other tests. Two different types of investment materials were selected, gypsum-and phosphate-bonded. Method 1 is a traditional three-point bending test. Twelve rectangular bars with dimension of (70 x 9 x 3 mm) were prepared and placed on supports 56.8 mm apart. The test was conducted at a cross head speed of 1 mm/min by use of a universal testing machine. The load applied to the test specimen and the corresponding deflection were measured until the specimen fractured. The E value was calculated from a linear part of the stress strain plot. Method 2 is an ultra micro-indentation system to determine near surface properties of materials with nanometer resolution. The measurement procedure was programmed such that the specimens were indented with an initial contact force of 5 mN then followed by a maximum force of 500 mN. Measurement consisted of 10 indentations conducted with a spherical stainless steel indenter (R = 250 microm) that were equally spaced (500 microm). The E value rose asymptotically with depth of penetration and would approach the three-point bending test value at approximately four time's maximum contact depth for both materials. Both methods are practical ways of measuring the E of investment materials. PMID- 15348006 TI - Physical and mechanical characteristics of a chlorine-substituted poly(para xylylene) coating on orthodontic chain modules. AB - A 10 microm film of a chlorine-substituted poly(para-xylylene) was evaluated as a protective coating for commercial chain modules. Segments of modules were either non-coated or coated using five modes: 0%, 100%, and 200% elongation during coating and shipping; and 100% and 200% elongation only during coating. Prior to hydration, coating coverage was examined via light and electron microscopy. When hydrated, the presence or absence of the coating did not appreciably change the total weight gain (1.5%). However, when the modules were post-coating elongated to either 100% or 200%, neither the non-coated nor the coated modules sorbed water. Hydration did not significantly change the mechanical properties of the modules. As expected, most of the mechanical properties of the coated modules were greater than the properties for the uncoated modules. With regard to the coated modules only the properties of those modules having 0% elongation during coating and shipping were, in general, significantly different from the other coated modules. Post-coating elongation reduced the stiffness of the coated modules more than 79%. In regard to stress relaxation, the non-coated and coated modules recovered approximately 74% and 62% of the peak loading, respectively. Using the load-time data from these curves, Maxwell-Weichert models accurately predicted the decay profiles of both the non-coated and coated modules. Because the water sorption and the mechanical properties of these coated modules were favorable, they should be further characterized for staining. PMID- 15348007 TI - Composition and structure of apatite formed on organic polymer in simulated body fluid with a high content of carbonate ion. AB - Apatite layer was formed on polyethyleneterephthalate (PET) substrate by the following biomimetic process. The PET substrate was placed on granular particles of a CaO, SiO2-based glass in simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma to form apatite nuclei on their surfaces. The apatite nuclei was then grown into a continuous layer by subsequently soaking the substrate in SBF under air or CO2 atmosphere in which CO2 partial pressure in the ambient was adjusted to 14.8 kPa to increase the content of carbonate ion to a level nearly equal to that of blood plasma. The increase in the content of carbonate ions in SBF changed the Ca/P atomic ratio of the apatite from 1.51 to 1.63, content of CO(3)2- ions from 2.64 to 4.56 wt %, and lattice constants a from 94.32 to 94.23 nm and c from 68.70 to 68.83 nm, respectively. The Ca/P ratio and lattice constants of the apatite formed in SBF under CO2 atmosphere were approximately identical to those of bone apatite, i.e. Ca/P atomic ratio 1.65, content of CO(3)2- ion 5.80 wt % and lattice constants a 94.20 and c 68.80 nm. This indicates that an apatite with composition and structure nearly identical to those of bone apatite can be produced in SBF by adjusting its ion concentrations including the content of carbonate ions to be equal to those of blood plasma. PMID- 15348008 TI - The processing and characterization of animal-derived bone to yield materials with biomedical applications: part 1: modifiable porous implants from bovine condyle cancellous bone and characterization of bone materials as a function of processing. AB - A study on the development of a process to form materials suitable for biomedical xenograft implants from bovine cancellous bone is presented. Bone cubes cut from the condyle portion of bovine femurs sourced from abattoir waste were subjected to a defatting and subsequent deproteination procedure to produce shape modifiable materials in which the biocompatible mineral calcium hydroxycarbonate apatite component was preserved in the original osseous architecture of the bovine bone. Optimum defatting was achieved by (1) thawing of the precut bone cubes in water, (2) pressure cooking at 15 psi in water, (3) soaking in 0.1 mol l(-1) NaOH followed by a thorough rinse under running water, (4) microwave heating of the bone cubes in water, (5) refluxing in methyl acetate and finally (6) removal of internal liquid from the cubes by shaking and then air drying. Subsequent deproteination of the defatted bone cubes was optimally achieved by (1) soaking in 5% sodium hypochlorite solution at ambient temperature using ultrasonication, (2) thorough rinsing of the cubes in water followed by drying. The final product is a defatted/deproteinated, bleached material that can be molded into various shapes for implant use in the body. The bone specimens were characterized by a suite of analytical techniques (i.e. infrared, 31P and 13C solid magic-angle spinning (MAS) nuclear magnetic resonance (NMR), X-ray photoelectron spectroscopies, atomic absorption (AA) spectrometry, inductively coupled plasma (ICP) spectrometry, differential scanning calorimetry (DSC), and scanning electron microscopy (SEM)) in order to follow compositional changes during the various stages of processing. In general, bovine condyles proved to be the best source of xenograft materials with condyles from other animal species (i.e. deer, sheep and ostrich) being too small to constitute a utilizable source of cancellous bone. This study shows how value can be added to a hitherto underutilized abattoir by-product by using simple processing techniques. PMID- 15348009 TI - Bone strains and anterior lift-off, measured with three alternative designs of tibial components of TKA. AB - Total knee replacement is a successful procedure with high clinical success rates. Problems are mostly initiated on the tibial side, and may be due to - amongst others - improper mechanical design of the tibial base plate. In this paper some new design concepts for the tibial component of a total knee prosthesis are presented. They are evaluated experimentally using a model for a proximal tibia, and strain gauge measurements and displacement measurements as experimental techniques. The designs are meant to yield a physiological load sharing between the trabecular and the cortical bone in the proximal tibia, and to minimize anterior lift-off of the tibial base plate. The optimal design required a metal backing of the plastic part and a thin continuous metallic rim in contact with the proximal tibial cortex. An optimal macro-composite structure within the plastic part was obtained by using thin steel wires in the transversal direction, connected to the metallic rim. With this optimal design, it was shown that the force required to close the anterior gap at simulated knee bending was smaller than 250 N, which can easily be applied clinically by an anteriorly placed clamp or bone screw. PMID- 15348010 TI - Hydroxyapatite formation on cellulose cloth induced by citric acid. AB - Hydroxyapatite (HAp) formation on cellulose cloth with the aid of citric acid was investigated. The cellulose cloths were soaked in simulated body fluid (SBF) solutions (1.5 SBF) with ion concentrations 1.5 times that of SBF (1.0 SBF) with and without citric acid and carbonate containing HAp crystals were found to form only in the 1.5 SBF solution that contained citric acid. The results were explained in terms of hydrogen bonding of citric acid to the cellulose cloth and its chelating ability of calcium ions. Practical application may involve the inclusion of citric acid in the 1.5 SBF solution to promote formation of HAp on previously bioinert cellulose cloth. PMID- 15348011 TI - Effect of temperature and immersion on the setting of some calcium phosphate cements. AB - Calcium phosphate cements based on powders containing alpha-Ca3(PO4)2 and aqueous solutions containing Na2HPO4 as accelerator were used to determine the effects of accelerator concentration, temperature and immersion on the setting time. Increases in accelerator concentration and temperature increased the rate of setting, but immersion had a retarding effect. These results were used to design a method whereby a syringe filled with cement paste can be kept ready for injection of the paste into the implantation site for a long time, whereas setting of the cement paste in the body takes place in a short time. PMID- 15348012 TI - The influence of chemical treatment and suture on the elastic behavior of calf pericardium utilized in the construction of cardiac bioprostheses. AB - Poor mechanical properties of biological tissue are known to cause wear, leading to the failure of cardiac bioprostheses made of calf pericardium. Different chemical agents such as sodium dodecyl sulfate (SDS) are presently being tested as possible inhibitors of the calcification process. The objective of this report was to determine the mechanical behavior of calf pericardium treated with SDS for 24 h and the influence of the suture on the mechanical properties of the tissue. Forty-eight samples were tested: 24 subjected to a standard treatment with glutaraldehyde (12 sewn with 4/0 silk suture thread) and 24 incubated with SDS for 24 h (12 sewn with the same suture thread). Each sutured and non-sutured sample was cut into two strips to yield paired samples. All were subjected to tensile stress to breaking point. The mean stress at breaking point in the non sutured series treated with glutaraldehyde alone was 16.42 and 13.85 MPa depending on the region of the pericardium, while in the sutured samples subjected to glutaraldehyde the mean stress was 7.50 and 7.63 MPa, respectively, differences which were statistically significant (p = 0.03 and p = 0.003, respectively) when the means for non-sutured samples from equivalent regions treated with glutaraldehyde were compared. The stress at breaking point was lower in the SDS-treated series, ranging between 2.60 and 3.56 MPa. The mathematical functions that govern the stress/strain or deformation were obtained. In the series of pericardium treated with SDS, deformations of 10% were produced with stresses of under 0.4 MPa, an outcome that is intolerable from the constructive point of view. We established a regression model that enabled us to determine the mechanical behavior of a sutured sample by testing a contiguous piece of tissue, with a high correlation coefficient (r ?gt 0.99). We consider this finding to be of interest in the selection of pericardium for use in the construction of leaflets for cardiac bioprostheses. PMID- 15348013 TI - Stress transmission through Ti-Ni alloy, titanium and stainless steel in impact compression test. AB - Impact stress transmission of Ti-Ni alloy was evaluated for biomedical stress shielding. Transformation temperatures of the alloy were investigated by means of DSC. An impact compression test was carried out with use of split-Hopkinson pressure-bar technique with cylindrical specimens of Ti-Ni alloy, titanium and stainless steel. As a result, the transmitted pulse through Ti-Ni alloy was considerably depressed as compared with those through titanium and stainless steel. The initial stress reduction was large through Ti-Ni alloy and titanium, but the stress reduction through Ti-Ni alloy was more continuous than titanium. The maximum value in the stress difference between incident and transmitted pulses through Ti-Ni alloy or titanium was higher than that through stainless steel, while the stress reduction in the maximum stress through Ti-Ni alloy was statistically larger than that through titanium or stainless steel. Ti-Ni alloy transmitted less impact stress than titanium or stainless steel, which suggested that the loading stress to adjacent tissues could be decreased with use of Ti-Ni alloy as a component material in an implant system. PMID- 15348014 TI - Quantitative analysis of the regulation of leukocyte chemosensory migration by a vascular prosthetic biomaterial. AB - The need for improved, infection-resistant vascular biomaterials calls for more objective evaluation of the immune pathophysiology of implantable prosthetic materials. In this study we have developed a new strategy to quantitatively characterize population-averaged responses of immune cell migration on vascular prosthetic materials. This approach, incorporating a chemokinetically regulated "biomaterial-gel" sandwich configuration, was applied to quantify both random and directed modes of the chemosensory migration of human neutrophil leukocytes on expanded polytetrafluoroethylene (ePTFE). Our studies show that (a) microporous, synthetic materials like ePTFE suppress the basal rate of random cell migration relative to that reported on non-porous control surfaces; (b) stimulation with chemoattractant (formyl peptide) can significantly elevate rates of random and directed migration on ePTFE; and (c) protein conditioning of ePTFE with albumin or immunoglobulin G can differentially modulate the rates and relative proportion of random and directional components of leukocyte migration response to chemoattractant. This, to our knowledge, is the first objective quantitation of chemokinetically regulated cell migration on implantable prosthetic materials. PMID- 15348015 TI - Processing and properties of two different poly (ortho esters). AB - Processing and properties of two structurally different poly (ortho esters) (POE) and their in vitro behavior have been compared. One POE was a copolymer of diketene acetal 3,9-diethylidene-2,4,8,10-tetraoxaspiro[5.5]undecane, trans cyclohexanedimethanol and polyacetal. The other POE was a copolymer of diketene acetal 3,9-diethylidene-2,4,8,10-tetraoxaspiro[5.5]undecane, trans cyclohexanedimethanol and trans-cyclohexanedimethanol-dilactide. The polyacetal POE was also used as a matrix incorporating two different model drugs, caffeine and furosemide. Both polymers were ultrasonically molded into slabs. The lactide modified POE was also melt-extruded into rods and self-reinforced using a solid state die-drawing technique. The initial strength values of both ultrasonically molded and extruded samples were close to those of non-oriented poly lactic acids. Self-reinforcing increased the shear strength, bending strength and modulus of the rods from 41.7+/-1.7 MPa to 66.7+/-2.3 MPa, from 78.5+/-22.8 MPa to 183+/-23 MPa and from 2.5+/-0.3 GPa to 4.4+/-0.3 GPa, respectively. In vitro hydrolysis of samples was studied either in phosphate buffer saline (PBS) or in simulated body fluid (SBF) at 37 degrees C. Samples degraded faster in unchanged PBS than in PBS changed weekly. The total in vitro strength retention time was relatively short varying from 14 days to 42 days depending on the processing method and polymer type. Samples retained their wet weight at initial level for a longer time but dry weight started to decrease already after the first in vitro week. Polymers or degradation products crystallized in SBF but remained amorphous when hydrolyzed in PBS. Some of the devices showed evidence of surface erosion, and the processing method affected the erosion. The storage time of these polymers was found to be limited. However, both polymers were found to be of interest in bone fixation and tissue engineering. PMID- 15348016 TI - The effect of storage in aqueous solutions on glass-ionomer and zinc polycarboxylate dental cements. AB - Cylindrical specimens (dimensions 6 mm diameter x 12 mm height) of glass-ionomer and of zinc polycarboxylate dental cement have been stored in aqueous solutions for periods of 24 h, 1 week and 1 month. The solutions were of varying composition and affinity for water, and storage in them resulted in fluctuations in mass of the cements, an effect which was attributed to differences in the partitioning of water between the solutions and the cement specimens. Unlike the zinc polycarboxylate, the glass-ionomer gained mass in most of the solutions examined (except Na(2)SO(4)), showing it to have a much greater affinity for water than the zinc polycarboxylate. Despite the fluctuations in water uptake by the glass-ionomer, and loss of water by the zinc polycarboxylate, no statistically significant differences in compressive strength were recorded in any solution at any storage time. This contrasts with results reported previously for zinc polycarboxylates using smaller specimens, showing that specimen size has an influence on the interaction of cements with storage solutions. PMID- 15348017 TI - Correlation between light intensity and exposure time on the hardness of composite resin. AB - This study evaluated the correlation between light intensity emitted by the curing units and exposure time on the of degree polymerization of dental composites by measured Knoop hardness. Specimens with 5 mm diameter and 2 mm height were prepared in a copper mold, covered with a mylar strip and polymerized for 30 sec by curing units with 50, 130, 180, 220, 280 and 520 mW/cm(2), respectively. The output for all units was measured using a Curing Radiometer (Demetron). Other specimens were polymerized for 45, 60, 75, 90, 105, 120, 135, 150, 165 and 180 sec utilizing units of 130, 220 and 280 mW/cm(2). Knoop hardness values of the top and bottom surfaces were measured after 24 h. The results indicate that the curing units with higher light intensity presented improved values of Knoop hardness on the top and bottom regions. As expected, the top surface always presented improved Knoop hardness values in relation to the bottom, after exposure for 30 sec. The values obtained on the surface were statistically superior compared to the values on the bottom (p<0.05) for the intensity of 130, 220 and 280 mW/cm(2) after 30, 45 and 60 sec of exposure. PMID- 15348018 TI - alpha-Tricalcium phosphate hydrolysis to hydroxyapatite at and near physiological temperature. AB - The kinetics of hydroxyapatite (HAp) formation by direct hydrolysis of alpha tricalcium phosphate (alpha-TCP) [alpha-Ca(3)(PO(4))(2)] have been investigated. Transformation kinetics were examined for reactions at 37 degrees C, 45 degrees C and 56 degrees C by isothermal calorimetric analysis. Setting times and morphologies of the resultant HAp were found to be strongly dependent on reaction temperature. XRD analysis accompanied by FTIR confirmed that phase pure calcium deficient hydroxyapatite (CDHAp) [Ca(10-x)(HPO(4))(x)(PO(4))(6-x)(OH)(2-x)] was formed. Complete reaction occurs within 18, 11, 6.5 h at 37, 45 and 56 degrees C, respectively. The extent of HAp formation differs for particulate slurries and pre-shaped forms of reactant alpha-TCP. Formation of hydroxyapatite in pre-formed pellets was hindered due to limited water penetration, but enhanced with the presence of NaCl as a pore generator. Regardless of the precursor characteristics and temperature, HAp formation is characterized by an initial period of wetting of the alpha-TCP precursor, an induction period and a growth period during which the bulk transformation to HAp occurs. The microstructures of the resultant HAp at all temperatures were generally similar and are characterized by the formation porous flake-like morphology. Microstructural coarsening was observed for the CDHAp formed above the physiological temperature. The hardening generated by the hydrolysis reaction was demonstrated using diametrical compression tests. The original tensile strength of 56% dense alpha-TCP increased from 0.70+/-0.1 MPa to 9.36+/-0.4 MPa after hydrolysis to CDHAp at 37 degrees C, corresponding to a density of 70%. PMID- 15348019 TI - Treatment of tooth fracture by medium energy CO2 laser and DP-bioactive glass paste: thermal behavior and phase transformation of human tooth enamel and dentin after irradiation by CO2 laser. AB - Acute trauma or trauma associated with occlusal disharmony can produce tooth crack or fracture. Although several methods are proposed to treat the defect, however, the prognosis is generally poor. If the fusion of a tooth fracture by laser is possible it will offer an alternative to extraction or at least serve as an adjunctive treatment in the reconstruction. The responses of soft tissues to lasers of different wavelengths are fairly well known, but the reactions of hard tissues are still to be understood. The purpose of this research was to study the feasibility of using a medium energy continuous-wave CO(2) laser and a low melting-point bioactive glass to fuse or bridge tooth fractures. The present report is focused on the first part of the research, the analysis of changes in laser-irradiated human tooth enamel/dentin by means of X-ray diffractometer (XRD), Fourier-transforming infrared spectroscopy (FTIR), differential thermal analysis/thermogravimetric analysis (DTA/TGA), and scanning electron microscopy (SEM). After CO(2) laser irradiation, there were no marked changes in the X-ray diffraction pattern of the enamel when compared to that before laser treatment. However, a small peak belonging to alpha-TCP appeared at the position of 2theta=30.78 degrees C. After being treated with CO(2) laser, the dentin showed much sharper peaks on the diffraction patterns because of grain growth and better crystallinity. alpha-TCP and beta-TCP were identified after laser treatment. In the FTIR analysis, an HPO(4)(-2) absorption band was noted before laser treatment disappeared after the irradiation. No significant change in the absorption band of HPO(4)(-2) was found on the FTIR curves of enamel after laser treatment. The results of DTA/TGA indicated that loss of water and organic materials occurred in both enamel and dentin after laser treatment. Under SEM, melting and resolidification occurred in both enamel and dentin by medium energy of CO(2) laser. This implies that using a continuous-wave CO(2) laser of medium energy density to fuse a low melting-point bioactive glass to the enamel/dentin is possible. We believe these phase changes and thermal data can make a useful guide for future studies on the thermal interaction and bridging mechanism between the bioactive glass and enamel/dentin under CO(2) laser irradiation. PMID- 15348020 TI - Surface induced reactivity for titanium by ion implantation. AB - Calcium and phosphorus storage in a thin layer of titanium surface was achieved by ion implantation. We study the reactivity of this surface in response to a hydrothermal treatment. The incipient implanted species are observed to convert to Ca(2+) and PO(4)(3-), the precursors for generating calcium phosphate polymorphs. Hydroxyapatite is formed from these precursors by an interface-liquid mediated mineralization preceded by the hydrolysis of oxygen compounds of Ca and P from the solid phase. The morphology and organization of apatite mineral is controlled by the fluid dynamics reflecting the surface remodeling to adapt to the available local environment. Exposed to calcium and phosphate ion containing solution, the hydrothermally treated surface templates hydroxyapatite deposition. Ca and P implanted Ti surface was shown to be chemically and morphologically actively involved in the interfacial reactions. PMID- 15348021 TI - Controlled release systems based on poly(lactic acid). An in vitro and in vivo study. AB - A new biodegradable delivery system based on poly(lactic acid) has been formulated, with potential applications in sustained antibiotic release against bone infection. The in vitro release of a new quinolone (pefloxacin) from low molecular weight poly(D,L-lactic acid) Mw = 2 x 10(3) lasted for 56 d whereas the in vivo delivery lasted 33 d. In both cases, the release rate is controlled by the drug diffusion and the polymer degradation, which seems to be the predominant factor. For the release experiments, discs were prepared from poly (D,L-lactide) Mw = 2 x 10(4) with drug loadings of 2% and 10% w/w. It was concluded that pefloxacin concentration remains higher than the Minimum Inhibitory Concentration (MIC) against the major causative bacteria of bone infection. The results indicate that the two different types of poly(lactic acid) can be used effectively in an implantable antibiotic release system. PMID- 15348022 TI - A novel technique for the detailed size characterization of wear debris. AB - The accurate and detailed characterization of artificial joint wear debris is important in determining both the wear rate of prostheses and understanding the role that the debris plays in the development and progression of aseptic loosening. The novel application of low angle laser light scattering (LALLS) to the particle size characterization of ultra high molecular weight polyethylene (UHMWPE) wear debris is described. The results demonstrate that both ex vivo and in vitro origin wear debris samples, at concentrations typical of those produced via an alkali-digestion retrieval route, can be reproducibly analyzed via LALLS. Because the LALLS route enables particle size analysis of the entire debris sample to be acquired non-destructively and whilst in suspension, artefacts associated with filtering, drying and agglomeration of debris are avoided, in contrast to currently used techniques such as filtration and scanning electron microscopy (SEM) observation. PMID- 15348023 TI - The influence of chemical treatment and suture on the elastic behavior of calf pericardium utilized in the construction of cardiac bioprostheses. AB - Poor mechanical properties of biological tissue are known to cause wear, leading to the failure of cardiac bioprostheses made of calf pericardium. Different chemical agents such as sodium dodecyl sulfate (SDS) are presently being tested as possible inhibitors of the calcification process. The objective of this report was to determine the mechanical behavior of calf pericardium treated with SDS for 24 h and the influence of the suture on the mechanical properties of the tissue. Forty-eight samples were tested: 24 subjected to a standard treatment with glutaraldehyde (12 sewn with 4/0 silk suture thread) and 24 incubated with SDS for 24 h (12 sewn with the same suture thread). Each sutured and nonsutured sample was cut into two strips to yield paired samples. All were subjected to tensile stress to breaking point. The mean stress at breaking point in the nonsutured series treated with glutaraldehyde alone was 16.42 and 13.85 MPa, depending on the region of the pericardium, while in the sutured samples subjected to glutaraldehyde the mean stress was 7.50 and 7.63 MPa, respectively, differences which were statistically significant (p=0.03 and p=0.003, respectively) when the means for nonsutured samples from equivalent regions treated with glutaraldehyde were compared. The stress at breaking point was lower in the SDS-treated series, ranging between 2.60 and 3.56 MPa. The mathematical functions that govern the stress/strain or deformation were obtained. In the series of pericardium treated with SDS, deformations of 10% were produced with stresses of under 0.4 MPa, an outcome that is intolerable from the constructive point of view. We established a regression model that enabled us to determine the mechanical behavior of a sutured sample by testing a contiguous piece of tissue, with a high correlation coefficient (r?gt 0.99). We consider this finding to be of interest in the selection of pericardium for use in the construction of leaflets for cardiac bioprostheses. PMID- 15348024 TI - Adhesion of microvascular endothelial cells to metallic implant surfaces. AB - The objective of this study was to explore the molecular mechanisms of adhesion of endothelial cells (ECs) to implant grades of titanium alloy (Ti) and stainless steel (SS), compared to tissue culture polystyrene (PS). The idea is that promotion of EC adhesion to implant surfaces during the initial stages of healing may be critical in the formation of a capillary bed intimately associated with the implant surface. Ultimately this could be expected in turn to promote bone formation close to the surface and a more stable implant/bone interface. Surfaces were coated with either peak 1 fibrinogen gammaAgammaA, fibrinogen Fr I-9, fibrinogen fragment D1, fibronectin, vitronectin, or fetal calf serum and then post-coated with bovine serum albumin (BSA) to block non-specific cell adhesion. Surfaces with BSA alone and no other protein coating were also evaluated. Fibronectin coating maximized cell adhesion on all three surfaces, and adhesion was highest on PS. BSA blocked cell adhesion to PS (and most adhesion to SS) much better than to Ti. These results provide evidence that BSA adsorption on the metal surface is unable to effectively block the adhesion of the cells to the Ti. These data may provide a basis for understanding in vivo observations that soft tissue becomes attached to a Ti surface more rapidly and with more bone formation than to SS. Evidence is also presented that alphavbeta3 plays an important role in adhesion of ECs to the Ti surface. These experiments also provide preliminary data which may reflect some of the features of initial EC adhesion to metal implants. PMID- 15348025 TI - Galvanic corrosion behavior of titanium implants coupled to dental alloys. AB - The corrosion of five materials for implant suprastructures (cast-titanium, machined-titanium, gold alloy, silver-palladium alloy and chromium-nickel alloy), was investigated in vitro, the materials being galvanically coupled to a titanium implant. Various electrochemical parameters E(CORR), i(CORR) Evans diagrams, polarization resistance and Tafel slopes) were analyzed. The microstructure of the different dental materials was observed before and after corrosion processes by optical and electron microscopy. Besides, the metallic ions released in the saliva environment were quantified during the corrosion process by means of inductively coupled plasma-mass spectrometry technique (ICP-MS). The cast and machined titanium had the most passive current density at a given potential and chromium-nickel alloy had the most active critical current density values. The high gold content alloys have excellent resistance corrosion, although this decreases when the gold content is lower in the alloy. The palladium alloy had a low critical current density due to the presence of gallium in this composition but a selective dissolution of copper-rich phases was observed through energy dispersive X-ray analysis. PMID- 15348026 TI - PMMA-based composite materials with reactive ceramic fillers: part III: radiopacifying particle-reinforced bone cements. AB - New acrylic bone cements were prepared from alumina particles previously treated by 3-(trimethoxysilyl)propylmethacrylate (gamma-MPS), able to act both as radiopacifying and reinforcing agents. The present study deals with the handling characteristics and the compressive behavior of such cements. The influence of the particles morphology, their surface-modification by gamma-MPS bonding agent, their concentration in the cement, the powder-to-liquid ratio and the benzoyl peroxide concentration are reported. The role of grafted gamma-MPS molecules as coupling agent was confirmed. For several formulations, compressive strength and modulus reached 150 MPa and 3400 MPa respectively. Limitations in the use of such formulations are also comprehensively discussed. PMID- 15348027 TI - Precipitation of (PdxAg1-x)3 In in Ag-25% Pd-In alloys and the partial phase diagram of ternary Ag-Pd-In alloys. AB - A study was carried out to determine the partial phase diagram for the system Ag 25 wt % Pd-y wt% In with y /= 0.83, the 24 h pHs of the slurries were 9 to 11, whereas for those with TTCP/DCPA of 0.10). Planar and volumetric anisotropies were compared to the isotropic condition (eta(sigma) = eta(epsilon) = 1.0) for all of the included studies. All of the studies reported cortical bone properties that were volumetrically anisotropic (p < 0.05), however, a common plane of isotropy was noted in the radial-circumferential (1-2) plane (p > 0.05). Future use of these parametric equations will allow further illucidation of the issue of mesomechanical and micromechanical levels of anisotropy within other tissues and materials of interest. PMID- 15348042 TI - The influence of simultaneous mechanical and thermal loads on the stress distribution in molars with amalgam restorations. AB - A finite element analysis (FEA) of a mandibular molar restored with Class II amalgam restoration was conducted to determine the stress distribution which results from a superposition of simultaneous mechanical and thermal loading. A fully crossed three-level four-factor experimental design was used to evaluate the relative influence of crown temperature, time of thermal loading, occlusal force, and cavo-surface margin adhesion on the stress distribution. It was found that occlusal force and temperature had significant influence on the stress distribution and particularly on the maximum principal stress. Over the range in oral conditions considered, thermal loading contributed for over 35% of the stress within the restored molar subjected to simultaneous mechanical and thermal loads. Furthermore, thermal loading had significant effects on the magnitude of normal stress that develops parallel to the pulpal floor. Although marginal bonding of amalgam reduces the stress resulting from occlusal forces, thermal loading promotes the development of significant interfacial shear stresses along the bonded margin. Stresses related to the thermal component of loading concentrate near the pulpal floor and lingual surface margin, the site most prominent in cusp fracture. Hence, results from this study clearly indicate that an evaluation of new dental materials and/or restorative designs should consider the effects from a superposition of simultaneous mechanical and thermal loads on fracture resistance. PMID- 15348043 TI - Proliferation/differentiation of osteoblastic human alveolar bone cell cultures in the presence of stainless steel corrosion products. AB - Human osteoblastic alveolar bone cells were cultured for 28 days in control conditions and in the presence of three non-lethal concentrations of AISI 316L stainless steel (SS) corrosion products. Cells were exposed to SS corrosion products in two experimental situations: (i) in selected stages of the incubation time (during the first, second, third and fourth week of culture); and (ii) during the 28 days incubation period. Cultures were characterized for cell proliferation, total protein content, alkaline phosphatase activity (ALP) and ability to form mineralized deposits; culture media was analyzed for ionized calcium (Ca) and phosphorus (P) concentrations throughout the incubation period. The presence of SS corrosion products during the different stages of the incubation period did not significantly affect the cell proliferation; however, a significant dose-dependent deleterious effect was observed on the levels and pattern of ALP activity, concentration of ionized Ca and P in the culture medium and, also, ability to form mineralized deposits, especially in cultures exposed during the first and second week of culture (respectively, lag phase and exponential cell growth phase). Similar effects were observed in cultures exposed to the SS corrosion products during the 28 days incubation period. However, the presence of such products during the third week (when the mineralization process occurs) and, also, during the fourth week, resulted in little or no significant effects on the behavior of alveolar bone cells. Results suggested that SS corrosion products above certain non-lethal concentrations may disturb the proliferation/differentiation relationship of osteoblastic human alveolar bone cell cultures. PMID- 15348044 TI - In vitro aging of a calcium phosphate cement. AB - Cement samples made of beta-tricalcium phoshate (beta-TCP), phosphoric acid (PA) and water mixtures were incubated in several aqueous solutions to determine their stability over time. The effects of the cement composition and the incubating temperature were investigated in more detail. The cement samples contained mostly dicalcium phosphate dihydrate (DCPD) and remnants of beta-TCP crystals. Depending on the initial cement composition, a certain amount of dicalcium phosphate (DCP) crystals were formed. The larger the initial PA concentration, the larger the DCP amount. After setting, the cement composition was stable for at least 16 days up to 60 degrees C. Above that temperature, the DCPD crystals decomposed into DCP crystals. The latter reaction provoked a decrease of the pH of the incubation solution, phenomenon expected for a cement sample containing an excess of PA. As the cement samples contained an excess of beta-TCP, it was postulated that beta TCP crystals became so covered by DCP or DCPD crystals during setting that the setting reaction was stopped prematurely. The latter phenomenon gave a good explanation for the low pH values measured in the incubation solutions. PMID- 15348045 TI - The prediction of polyethylene wear rate and debris morphology produced by microscopic asperities on femoral heads. AB - Counterface damage in the form of scratches, caused by bone cement, bone or metallic particles, has been cited as a cause of increased wear of ultra-high molecular weight polyethylene (UHMWPE) acetabular cups. It is known that high levels of particulate wear debris lead to osteolysis. Surface damage was characterized in a series of explanted Charnley femoral heads. The heads had a mean scratch height of 1 microm with a mean aspect ratio (defined as height divided by half width) of 0.1. Wear discs were artificially scratched using these scratch geometries as a guide. In addition, the scratch geometries were incorporated into a finite element model of a stainless steel asperity repeatedly sliding over UHMWPE under conditions similar to those in an artificial hip joint. Wear tests showed a strong correlation between the average cross-sectional area of the scratch lip above the mean zero line and the measured wear factor. The finite element model predicted increases in the area of UHMWPE suffering plastic strain with increases in the cross-sectional area of the asperity above the mean line. Analysis of the wear debris showed the mode of the particle size was 0.01 0.5 microm for all cases. The morphology of the particles varied with aspect ratio of the asperity, with an increased percentage mass of submicrometer-sized debris with increased scratch lip aspect ratio. The finite element results predicted that the maximum surface strains would increase with increasing asperity aspect ratio. Examination of the worn UHMWPE pin surfaces showed an association between increased surface damage, probably due to high surface strains, and increased aspect ratio. The large areas of surface plastic strain predicted for asperities with high cross-sectional areas above the mean line offer an explanation for the positive correlation between wear rate and the average cross-sectional area of the scratch lip material. The higher surface strains predicted for the higher aspect ratios may explain the increased percentage mass of biologically active submicrometer-sized wear particles found for scratch lips with higher aspect ratios. PMID- 15348046 TI - Expression of heat shock protein 70 (HSP70) at the interface of polymer-implants in vivo. AB - Synthetic polymer meshes are widely applied in the modern surgical approach for repairing abdominal wall defects. The implanted material is often observed leading to post-operative complications such as deficient abdominal wall mobility and adhesion formation with the abdominal cavity and/or abdominal organs. However, the functioning of the implant is primarily affected by the wound healing process guided by inflammatory events occurring at the tissue-material interface. This could presumably be influenced by the physicochemical properties of the polymer. With regard to it, the cellular and molecular processes involved in the successful restoration of the abdominal wall function are poorly understood. The present in vivo study, therefore, exemplary investigated in a rat model, the commercially available polymer-meshes Prolene (polypropylene, PP), Mersilene (polyester, PE) and Vicryl (polyglactin 910), as well as new mesh variants consisting either of PP (EB) or a combination of PP and polyglactin 910 (A plus or Vypro). The implanted material was evaluated by light and electron microscopy, immunohistochemistry as well as morphometry over an implantation period of 90 days. The data show that polymers induce heat shock protein (HSP)70, and its expression at the interface correlates inversely with the activity of the inflammatory reaction in vivo. Further, an ascent in HSP70 expression parallels the increasing implantation period and evolving foreign-body granulomas. Accordingly, a major role for HSP70 in modulating the local acceptance of polymers and as an additional marker for in vivo testing of polymers is suggestive. PMID- 15348047 TI - Morphology and immersion behavior of plasma-sprayed hydroxyapatite/bioactive glass coatings. AB - A series of hydroxyapatite/bioactive glass (HA/BG) coatings have been plasma sprayed on Ti6Al-4V substrate using HA/BG powders that were prepared by both sinter-granulation and direct mixing methods. The morphology and immersion behavior of these coatings in a simulated body fluid (SBF) were investigated. The results showed that in-house fabricated BG and sinter-granulated HA powders were irregularly shaped and dense. When 5 wt % or more BG was added in HA, the powder became rough and porous. X-ray diffraction (XRD) patterns showed that the presence of BG enhanced the decomposition of HA structure during fabrication of the powders. Reasonably high bond strengths were obtained from all coatings. The granulated type HA/BG coatings showed no significant differences in bond strength from the mixed type HA/BG coatings. The plasma spray process itself and the presence of BG enhanced the decomposition of apatite. Surface morphology of all sinter-granulated type coatings was similar to that of monolithic HA coating, that was comprised of patches of smooth and shiny glassy film and irregularly shaped particles on its surface. The dissolution depth of plasma-sprayed coatings immersed in SBF was largely dependent on the type and composition of the coating. Granulated type HA/BG coatings were much less dissolvable than monolithic HA or mixed type HA/BG coatings. It seems that the presently used granulation method for the preparation of HA/BG powders plays a predominant role in determining the dissolution behavior of the plasma-sprayed coatings. PMID- 15348048 TI - Analysis of rat plasma proteins desorbed from gold and methyl- and hydroxyl terminated alkane thiols on gold surfaces. AB - It is believed that adsorbed blood or plasma components, such as water, peptides, carbohydrates and proteins, determine key events in the concomitant inflammatory tissue response close to implants. The aim of the present study was to develop a procedure for the collection and analysis of minor amounts of proteins bound to solid metal implant surfaces. The combination of a sodium dodecyl sulfate washing method coupled with a polyacylamide gel electrophoretic protein separation technique (SDS-PAGE), Western blot and image analysis enabled the desorption, identification and semiquantification of specific proteins. The analyzed proteins were albumin, immunoglobulin G, fibrinogen and fibronectin. Concentration procedures of proteins were not required with this method despite the small area of the test surfaces. The plasma proteins were adsorbed to pure gold and hydroxylated and methylated gold surfaces, which elicit different tissue responses in vivo and plasma protein adsorption patterns in vitro. The image analysis revealed that the pure gold surfaces adsorbed the largest amount of total and specific proteins. This is in accordance with previous ellipsometry/antibody experiments in vitro. Further, the principles described for the protein analysis can be applied on implant surfaces ex vivo. PMID- 15348049 TI - An in vivo study on the reaction of hydroxyapatite-sol injected into blood. AB - In order to identify the possibility of hydroxyapatite-sol being used as a drug carrier and absorbent, an in vivo experimental study was performed. Pure hydroxyapatite microcrystals were synthesized by reaction of high purity Ca(OH)2 and H3PO4 solutions while using an ultrasonic homogenizer. Hydroxyapatite-sol was prepared by dispersing hydroxyapatite microcrystals into physiological salt solution. The hydroxyapatite-sol in different concentrations was injected into veins of both 25 Wistar rats and 5 Beagle dogs. The medium lethal dose was determined as 160 mg/kg. By observing the change of O2 and CO2 gas partial pressure, it was considered that the main cause of death by hydroxyapatite-sol injection was due to the blockage of capillaries. When one-sixth amount of the medium lethal dose was injected into the veins of the dogs, the value of phosphorous increased but calcium and magnesium kept stable. LDH, CPK, GOP and GDT values dramatically increased in 30 min after injection, however, one day after injection, the values returned to normal. Repeated experiments by similar methods were continued on same animals for 2 years in two-week intervals, the results in every experiment were almost same, no chronic damage or permanent side effects were discovered in the two years experiment. According to the results above, it was suggested that the hydroxyapatite-sol could be applied as a drug carrier into blood by using a small amount less than one-sixth of the medium lethal dose. PMID- 15348050 TI - Improvement of titanium alloy for biomedical applications by nitriding and carbonitriding processes under glow discharge conditions. AB - Although titanium alloys are used in medicine, they present low wear resistance. In this paper we present the results of studies on surface layers produced by nitriding at three different temperatures, and by carbonitriding under glow discharge conditions in order to improve wear resistance, hardness, and to modulate microstructure and chemical composition of surface layers. A cell culture model using human fibroblasts was chosen to study the effect of such treatments on the cytocompatibility of these materials. The results showed that nitrided and carbonitrided surface layers were cytocompatible. Modulation of surface microstructure by temperature in the nitriding process and chemical composition of surface layers by carbonitriding led to differences in cellular behaviour. Cell proliferation appeared to be slightly reduced from the 6th day of culture on nitrided surfaces produced at 730 degrees C and 1000 degrees C, however after 12 days of culture, the best growth was on surface layers produced at 850 degrees C. The best viability was observed on the carbonitrided layer. The orientation and shape of the cells corresponded to surface topography. Nitriding and carbonitriding under glow discharge conditions may constitute interesting techniques allowing the formation of surface layers on parts with sophisticated shapes. They may also permit modulating surface topography in a way improving the features of titanium alloys for various applications in medicine. PMID- 15348051 TI - Healing of titanium implants in onlay bone grafts: an experimental rabbit model. AB - An experimental rabbit bone graft model for the study of bone formation and remodeling around titanium implants is described. A 2.5-cm long radius bone segment served as an onlay graft. Two commercially pure (c.p.) titanium implants were inserted into the bone graft prior to fixation to the inferior border of the mandibular base with osteosynthesis titanium screws. Each animal was operated twice, allowing follow-up periods of 6 weeks on one side and 6 months on the contralateral side. In order to study bone remodeling by means of fluoroscopy the animals received single injections of tetracyclin and alizarine complexone 2 weeks and 1 week, respectively, prior to sacrifice by perfusion fixation with glutaraldehyde. The bone and implants were excized en bloc, postfixed and embedded in plastic resin. Stained and unstained thin ground sections as well as microradiographed thick sections were produced for light microscopic morphometry and fluoroscopy. After 6 weeks, osteoclastic/osteoblastic activity was primarily observed in the graft-recipient contact area and in the intracortical compartment of the graft bone. New bone formation observed on the implant surface originated from the recipient site. The bone formation was evident also in the implant-graft interface. At 6 weeks the average bone fill of the implant threads was 28.4% which increased to 36.4% after 6 months as measured by morphometry. An average of 17.6% bony contact was measured after 6 weeks which increased to 29.7% 6 months after surgery. The graft bone had reduced in size from an average of 39.5% after 6 weeks down to 24.8% after 6 months (P ?lt 0.05). It is concluded that the described experimental model can serve as a useful method for the study of implant healing in onlay grafts. PMID- 15348052 TI - Surface design of orthopaedic drug delivery implants: X-ray photoelectron spectroscopy of bone-derived apatites. AB - Incorporation of foreign ions onto a biomineral surface as well as removal from it of native (physiological) ions during deproteination procedures, are discussed in view of surface design of orthopaedic implants serving as drug delivery carriers. Surface properties of bone-derived apatites, purified at low temperature in non-aqueous medium, and of several commercial samples are examined via XPS technique. It has been demonstrated that, in spite of the fact that the initial bone demineralized matrix is derived from the same bovine bone, different deproteination procedures lead to different surface properties. Correlation between carbon 1s deconvoluted peak and the percentage of the residual organic fraction on the bone-derived apatites' surface at different stages of purification has been established. PMID- 15348053 TI - Load-bearing behavior of a simulated craniofacial structure fabricated from a hydroxyapatite cement and bioresorbable fiber-mesh. AB - Calcium phosphate cements (CPC) have proven successful in the repair of small, non-stress bearing skeletal defects. These cements do not have sufficient tensile strength or fracture toughness to allow their use in stress-bearing applications. It was hypothesized that a bioresorbable fiber mesh would improve the load bearing behavior of shell structures fabricated of CPC. This study used a biaxial flexure fixture to compare the work-to-fracture values of discs made of: (1) CPC; (2) CPC reinforced with a bioresorbable two-dimensionally oriented poly(glactin) fiber-mesh; and (3) poly(methyl methacrylate) (PMMA) that were immersed in a serum-like solution for 0-28 days. CPC-mesh and PMMA discs were indistinguishable at 0, 1 and 7 days, based on work-to-fracture data. CPC and CPC-mesh discs were indistinguishable at day 28, because of fiber hydrolysis. The knitted fiber-mesh was effective in improving load-bearing behavior of a calcium phosphate cement for potential structural repair of bone defects. PMID- 15348054 TI - Mechanical and fracture behavior of a fiber-reinforced bioabsorbable material for orthopaedic applications. AB - Tensile and fracture tests were conducted on thin panels of a fiber-reinforced bioabsorbable material. The composites were made of polycarbonate matrix and calcium phosphate fibers. Both matrix and fibers were bioabsorbable orthopaedic biomaterials. The fibers were short and randomly distributed. The properties were compared for composites with and without a coating of methane on the fibers. Composites with the methane coating had a higher elastic modulus, a higher proportional limit, but a lower load at failure on pre-cracked panels. Electron microscopy showed that the coating resulted in a better bond between the matrix and the fiber. PMID- 15348055 TI - Effect of several additives and their admixtures on the physico-chemical properties of a calcium phosphate cement. AB - Combinations of citrate (C6H5O(7)3-), pyrophosphate (P2O(7)4-) and sulfate (SO(4)2-) ions were used to modify the physico-chemical properties of a calcium phosphate cement (CPC) composed of beta-tricalcium phosphate (beta-TCP) and phosphoric acid (PA) solution. The results obtained with only one additive at a time are similar to those previously published. New facts are: the positive effect of C6H5O(7)3- ions on cement failure strain and their negative effect on cement pH. The position of the setting time maximum measured at an SO(4)2- concentration of 0.09 M was not displaced by the addition of C6H5O(7)3- and P2O(7)4- ions. However, the effect of SO(4)2- ions on the setting time was depressed by C6H5O(7)3- ions. Moreover, no increase in tensile strength was observed when increasing amounts of SO(4)2- were added into a C6H5O(7)3- containing cement. The latter results suggest a competitive effect of C6H5O(7)3- and SO(4)2- on setting time and tensile strength. Anhydrous dicalcium phosphate (DCP; CaHPO4) appeared in cement samples dried just after setting, but not in cement samples incubated for 24 h in deionized water before the drying step. It is believed that the setting reaction is stopped by the drying step, leaving a low internal pH in the sample, hence providing favorable conditions for the transformation of dicalcium phosphate dihydrate (DCPD) into DCP. Interestingly, even though C6H5O(7)3- ions dramatically lowered the equilibrium pH of the cement with 5 ml of deionized water, they still prevented the occurrence of the transformation of DCPD into DCP. PMID- 15348056 TI - Quantitative analysis of polyethylene wear debris, wear rate and head damage in retrieved Charnley hip prostheses. AB - Submicrometer- and micrometer-sized ultra-high molecular weight polyethylene (UHMWPE) wear particles have been associated with osteolysis and failure of total artificial joints. Previous studies have isolated predominantly submicrometer sized particles at the expense of larger particles (>10 microm). This study aimed to isolate and characterize quantitatively all sizes of UHMWPE wear particles generated in 18 Charnley hip prostheses. In addition, to analyze the wear debris with respect to the total volumetric wear of the cup and damage to the femoral head. Particle size distributions ranged from 0.1 to ->1000 microm. A significant proportion (3-82%) of the mass of the wear debris isolated was>10 microm. The mode of the frequency distribution of the particles was in the range 0.1-0.5 microm for all patients. However, analysis of the mass of wear debris as a function of its size allowed differentiation of the wear debris from different patients. Femoral head damage was associated with high volumetric wear and increased numbers of biologically active submicrometer-sized particles. PMID- 15348057 TI - A new quantitative test method for cell proliferation based on detection of the Ki-67 protein. AB - A quantitative method to assess cell proliferation is one essential prerequisite for testing biomaterial cytocompatibility in vitro. Currently used methods, e.g. bromodeoxyuridine incorporation, show serious disadvantages concerning either sensitivity, specificity or handling. A new enzyme linked immunosorbent assay (ELISA) system for the quantification of cell proliferation based on detection of the Ki-67 protein is described. This protein has turned out to be strictly correlated with the active parts of the cell cycle but to be absent in G0. The measurement of Ki-67 expression by different human cell types, e.g. endothelial cells and HeLa cells, was evaluated in order to answer the question of whether the data obtained using the Ki-67 ELISA method correlate with the proliferation measured with flow cytometrical DNA analysis and microscopical evaluation. Methods currently used for the evaluation of cell proliferation were compared to the new Ki-67 ELISA method. In addition, the functionality of adherent endothelial cells, and the viability and morphology of the cells were investigated. Cells were treated with standard culture medium with or without the transcription inhibitor, actinomycin D, or growth factors, e.g. endothelial cell growth factor (ECGF), and were exposed to metal ion standard solutions. These solutions were in a cytotoxic-non-cytotoxic range. Ki-67 ELISA was found to be a reliable quantitative method to assess proliferation of cultured human cells in vitro. It has advantages over methods that are currently being used. It is easy to perform and corresponds to the requirements for a test to be selected for biomaterial testing according to ISO standard 10 993. PMID- 15348058 TI - Porous ceramic bodies for drug delivery. AB - An approach to the production of ceramic drug delivery devices is proposed. Two examples of possible ceramics are dealt with: hydroxyapatite weakly modifiable by living tissue and the bioinert alumina. The possibility to control the formed porosity was taken into consideration for both materials. The ratio between the acquired porosity and the quantity and quality of the agents inducing porosity is also described and discussed. A test on the role of porosity was performed on the obtained porous ceramic bodies and a study was made on the release of a substance with pharmacological activity from previously impregnated porous ceramic bodies. This paper is preliminary to a planned work targeted to the preparation of ceramic drug delivery systems. PMID- 15348059 TI - Cellular responses to chitosan in vitro: the importance of deacetylation. AB - Chitin and chitosan (a deacetylated derivative of chitin) have been proposed for biomedical applications because of their biocompatibility and abundance in nature. We have investigated the effect of the percentage of deacetylation (%DD) of chitosan on biocompatibility from two sources, shrimp and cuttle fish, with two cell lines, L929 and BHK21(C13). The difference in %DD for each source was approximately 10% in the range of 76-90%. Biocompatibility was investigated for: (1) cell adherence and growth on the chitosan samples as substrate; (2) the effect of extract media on 2d and 7d growth; and (3) the presence of an inhibition zone. The results were similar for both cell lines. The chitosan samples were air-dried on to tissue culture-grade petri dishes to provide a substrate for the adherent-cell cultures. The higher %DD substrates from each source supported attachment of the cells, while the lower %DD did not. Cells cultured in medium conditioned by each substrate (i.e. extract medium) displayed an initial difference in growth which was abrogated in cultures incubated for 7 days. No inhibition zone was apparent. However, after 7 days, some cells were noted migrating on to the low %DD substrate disks. The morphology of these cells was changed with the presence of pseudopodia being apparent. Thus, especially with regard to attachment the %DD has a very important effect on the biocompatibility of the chitosan and should be monitored carefully. PMID- 15348060 TI - Chemical transformation of some biologically relevant calcium phosphates in aqueous media during a steam sterilization. AB - The purpose of this study was to investigate the effect of steam sterilization on some biologically relevant calcium phosphates: CaHPO4 . 2H2O (DCPD), calcium deficient apatite (CDA) and biphasic calcium phosphate (BCP). Suspensions of 0.2 g of each calcium phosphate compound with 5.0 ml of deionized water were prepared and steam sterilized in an autoclave (20 min at 121 degrees C). After sterilization the suspensions were filtered and the dried solids characterized with scanning electron microscopy, IR-spectroscopy and X-ray diffraction. The pH and calcium concentrations of the filtrates were determined with ion selective electrodes. Similar measurements were made with the same samples which were not sterilized. The sterilization procedure was found to result in the dehydration of DCPD and hydration of calcium oxide incorporated into the BCP. Solution pH was observed to change from 7.3 to 5.5 for the solutions in equilibrium with DCPD and from 8.5 to 10.6 for those in equilibrium with BCP. Minor changes both with the solid and liquid phases were found to occur during the steam sterilization of CDA. These results indicate that steam sterilization may have different effects on different calcium phosphate suspensions: it can result in dehydration of DCPD, fast hydration for CaO in BCP, but no significant effect on CDA. PMID- 15348061 TI - Dissolution behavior of plasma-sprayed hydroxyapatite coatings. AB - The long-term stability of plasma-sprayed hydroxyapatite coatings is influenced by the dissolution behavior of the coating in in vivo conditions. Plasma-spraying generates a mixture of phases and this study has focused on how the balance of phases affects the in vitro dissolution behavior of the coatings in double distilled-deionized water and in tris-buffer solutions. The pH changes in double distilled-deionized water were monitored, whilst the pH value was maintained at 7.25 for the tris-buffer solution at 37 degrees C with 5% CO2 atmosphere. The phosphate and calcium ions released were measured using UV-Visible Spectrophotometer and Atomic Absorption Spectroscopy respectively. Changes in crystal and surface topology were also studied. The results indicate that the dissolution behavior of the coatings depends on several factors. The rate of release of phosphate ions was found to increase significantly for the tris-buffer solution compared to the deionized water, indicating that the presence of electrolyte constituents affects the dissolution behavior of the coatings. The Ca/P ratio in the tris-buffer solution is approximately three. Increases in the level of crystallinity of the coatings significantly decreased the dissolution rate and hence, the amount of phosphate ions released. The higher the percentage of crystallinity, the higher the stability of the coating under in vitro conditions. PMID- 15348062 TI - Biomechanical and biological properties of the implant material carbon-carbon composite covered with pyrolytic carbon. AB - The aim of this study was to test C/C material (carbonized, graphitized or covered with pyrolytic carbon) designated for the use in orthopaedic and bone surgery. Using an in vitro assay we confirmed, that the cell proliferation was exhibited the mostly on the C/C composite coated with pyrolitic carbon and afterwards polished. The two latest of subsequent water extracts of this material had a slightly inhibiting effect on the cells metabolic activity. Biocompatibility test in vivo performed subcutaneously on rats did not show big differences between three tested implants (C/C composite, epoxy resine, titanium alloy), on the other hand the plates tested on pigs demonstrated foreign-body reaction induced by wear C/C composite material. Such debris were found both in the neighborhood of the implant as well as in the lymphatic node. PMID- 15348063 TI - Characterization of the transformation from calcium-deficient apatite to beta tricalcium phosphate. AB - The structural changes that occur during the transformation of a Ca-deficient apatite, prepared by a wet chemical method, to beta-TCp were investigated. X-ray diffraction (XRD) analysis of as-prepared samples and samples calcined at temperatures between 500 and 1100 degrees C showed that the transformation occurs over the temperature range 710-740 degrees C, under non-equilibrium conditions. The change in crystallite size with increasing calcination/sintering temperature was studied by XRD using the Scherrer formula. Fourier transform infra-red (FTIR) analysis indicated considerable structural change in samples above and below this temperature range. Changes were observed in the hydroxyl, carbonate and phosphate bands as the calcination temperature was increased from 500 to 1100 degrees C. Even once a single beta-TCP phase is obtained at 740 degrees C there remains a considerable amount of structural change at temperatures between 740 and 1100 degrees C. This effect was illustrated by an unusual change in the lattice parameters of the beta-TCP structure and significant changes in the phosphate bands of FTIR spectra as the calcination temperature was increased. The results obtained in this study show that the combined experimental techniques of XRD and FTIR are excellent complimentary methods for characterizing structural changes that occur during phase transformations. PMID- 15348064 TI - Surface topography and HA filler volume effect on primary human osteoblasts in vitro. AB - HAPEX, a bone analog material, with similar properties to cortical bone, has been studied in vitro with particular reference to the effect of surface topography. The stimulation of a favorable bone response by this composite depends on optimization of the hydroxyapatite (HA) content in relation to the material bioactivity without compromising the mechanical characteristics. In this study we have started to investigate the effects of surface topography on cell attachment and subsequent cellular behavior in relation to proliferation. Different volumes of HA (20% and 40%) were added to a high density polyethylene (HDPE) matrix to produce the test materials. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to examine cell morphology on HAPEX, and the surface characteristics produced by different machining protocols. The measurement of cellular DNA and tritiated thymidine ([3H]-TdR) incorporation has been used to asses cell proliferation upon the materials. The results show that the material surface topography has a large influence on cell proliferation and attachment, and with a controlled material topography the 40% vol HA/HDPE composite gives the greater biological response compared to the 20% vol HA/HDPE composite. PMID- 15348065 TI - Tissue reaction against a self-setting calcium phosphate cement set in bone or outside the organism. AB - Calcium phosphate cements are able to set in situ when injected into bone tissue. We evaluated the tissue reaction occurring when a DCPD-based calcium phosphate cement was either set within the bone or implanted when already set. The samples were implanted in rabbit condyles and examined histologically after 8 and 16 weeks. The relative bone surface, the fibrous capsule around the implants and the implant section surface were measured. Solid material seemed to be better tolerated than paste implants. More bone was found at the solid implant contact whatever the implantation time and the solid material degraded much less rapidly. In conclusion, the physico-chemical modification of the biological environment occurring during setting increases the foreign body reaction against the material. PMID- 15348066 TI - beta-Chitin-based wound dressing containing silver sulfurdiazine. AB - Physical and biological properties of some wound dressing materials based on beta chitin were studied. Water vapor transmission rates (WVTR), oxygen permeabilities and biodegradation kinetics were examined for film-type samples. WVTR of samples was in the range 2400-2800 g/m2/day. However, oxygen permeabilities of the samples were relatively low. To improve oxygen permeabilities, porous sponge-type wound dressing materials were prepared. In addition, these sponge-type samples contained antimicrobial agents, silver sulfadiazine (AgSD), in order to prevent bacteria infection on a wound surface. Anti-microbacterial tests on agar plate were carried out to confirm the bactericidal capacity of present materials. These materials impregnating AgSD had the complete bactericidal capacity against pseudomonas aeruginosa up to 7 days. Finally, a wound healing effect of beta chitin-based semi-interpenetrating polymer networks was evaluated from the animal test using the wistar rat in vivo. Histological studies confirm the proliferation of fibroblasts in the wound bed and a distinct reduction in infectious cells. PMID- 15348067 TI - The protective efficacy of surgical latex gloves against the risk of skin contamination: how well are the operators protected? AB - Latex gloves are used by surgical staff to avoid exposure to patient body fluids, thus reducing the risk of contracting bloodborne viral diseases, such as hepatitis C and HIV. We studied the efficacy of the surgical barrier provided by latex gloves, before and after use in the operating theater. The electrical conductivity, insulation and mechanical resistance of glove latex were investigated, using routine supplies of surgical gloves. Latex structure was assessed by scanning electron microscopy and by mercury intrusion porosimetry. Latex is subject to hydration, a phenomenon associated in the laboratory with the loss of its electrical insulation properties. Such glove latex properties were found to be highly variable, with latex hydration times varying between 2 and more than 30 min. Rapidly hydrating gloves showed increased permeability to methylene blue, associated with higher levels of porosity. Thirty min of surgical use was associated with measurable hydration of glove latex and a statistically significant loss of electrical and mechanical resistance, with rupture load decreasing by 24%. Electronic control of the insulation properties of gloves during surgery permits early detection of hydration, and allows prompt correction by glove change, before the gloves lose their electrical and mechanical competence. PMID- 15348068 TI - Determination of quantitative leucite content in pressable ceramics compared to conventional dental porcelains. AB - In the present study, the leucite content of two different veneering porcelains and one pressable ceramic have been determined by X-ray diffraction after establishing a standard, using artificial leucite. The results are directly linked to the microstructure and mechanical properties of the materials. The most homogeneous glass-ceramic exhibits the highest relative leucite content and the highest fracture strength. PMID- 15348069 TI - Reliability of Pd based dental alloys regarding to corrosion resistance and production processes. AB - The knowledge of the relationship among chemical composition, microstructure, productive processes, functional properties, quality and price of the final product is an ever greater requirement for the firms producing dental alloys as well as for the dental laboratories making prosthesis. The present study deals with the evaluation of the corrosion resistance of high-Pd and Pd-Ag alloys by quantification of released ions following the recent European regulations suggested. Moreover the reliability of the alloys as far as concern their corrosion resistance was also investigated in relation to the different steps of alloy industrial production and restoration forming processes as well as after homogenization treatments followed by different cooling rate. The very complex microstructure of both alloys was greatly affected by the different casting conditions, deformation rate and thermal treatments used in alloy industrial production and prosthesis manufacturing. However the corrosion resistance of both alloys remained very good in all metallurgical states. The potentiodynamic tests allowed to distinguish among the structural conditions: the homogenization treatments, eliminating segregation inside the different phases, improved further the corrosion resistance of both alloys. PMID- 15348070 TI - The influence of poly(acrylic acid) molar mass on the properties of polyalkenoate cements formed from zinc oxide/apatite mixtures. AB - The influence of poly(acrylic acid) molar mass was investigated on cements formed from zinc oxide-apatite mixtures at three aging times; one, seven and 28 days. Cements based on both hydroxyapatite and fluorapatite were investigated. The compressive strength, un-notched fracture strength and fracture toughness increased markedly with poly(acrylic acid) molar mass. The fracture toughness and un-notched fracture strength increased with aging time for the two highest molar mass cements, but decreased with time for the two lowest molar mass cements. The greater chain entanglement density present in the higher molar mass cements is thought to contribute significantly to the cement stability in addition to the crosslinking of the polyacrylate chains by calcium and zinc ions. Substitution of hydroxyapatite by fluorapatite had no significant influence on the mechanical properties of the cements at aging times longer than one day. PMID- 15348071 TI - A study of the process and kinetics of electrochemical deposition and the hydrothermal synthesis of hydroxyapatite coatings. AB - Hydroxyapatite (HAp) coatings were prepared using electrochemical deposition and post-hydrothermal synthesis. The composition and morphology of coatings at each processing step was studied through the application of scanning electron microscopy (SEM), X-ray diffraction (XRD) and infra-red spectroscopy (IR). The mechanism and kinetics of hydrothermal synthesis were considered in particular, and the influence of the temperature and time on the HAp formation rate was also investigated. The results show that the electrochemical deposition coatings are composed of CaHPO42H2O crystals which are converted into needle-like HAp crystals after post-hydrothermal treatment. The HAp content of the coatings increases with the treatment temperature and time. The synthesis rate also increases with the pH value of the water. The formation of HAp coatings is considered to be a combination of several reactions. An Arrhenius relationship was found between the HAp formation rate and the temperature, and an apparent activation energy of 94.4 KJ/mol was obtained by calculation. PMID- 15348072 TI - On the heterogeneous structure of thermally cured bis-GMA/TEGDMA resins. AB - In this paper we report on the morphogenesis of dental resins formed by a thermally initiated radical cross-linking copolymerization of model dental resins. These systems were considered reasonable model simplifications of the actual resins used in commercial filling composites, veneers, etc. Effect of bis GMA content varying from 0 to 100% on the degree of conversion and morphology of the cured resin was investigated. Dynamic scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), dynamic mechanical thermal analysis (DMTA) and high temperature solvent extraction experiments were performed in order to determine the nature and location of unreacted unsaturations in relation to bis-GMA concentration. The interval of cure temperatures, ranging from 23 to 250 degrees C, may seem irrelevant from the clinical point of view but is instrumental for an understanding of the processes involved. Single glass transition temperature (Tg) found in DMTA experiments showed that the copolymerization process between bis-GMA and TEGDMA is random in nature, however, the overall cure reaction is inhomogeneous and this inhomogeneity is increasing with increasing bis-GMA molar fraction. Heterogeneous character of the cure was reflected into the network morphogenesis characterized by microgellation preceded by cyclization and, following connection of microgels. These cure kinetics resulted in a heterogeneous morphology of the cured resin. Moreover, it was shown that the cure kinetics controls to a great extent the degree of conversion of the reactive vinylidene C=C bonds in a fully cured resin. A decrease in bis-GMA concentration in the resin mixture led to a higher degree of conversion, however, even a neat TEGDMA homopolymer did not exhibit C=C conversion greater than 65-70% under the cure conditions used. Leaching out tests, consisting of an extraction of unreacted monomer from a finely ground cured resin in boiling cyclohexanone at 160 degrees C over a period of 5 d, revealed total weight loss of the order of 3.2-5.6 wt % regardless of the total conversion. No uncured monomer was detected in the extract solution using FTIR spectroscopy, however, the method used is not sensitive to the presence of oligomers. It was concluded that the remaining unreacted double bonds are the inevitable result of the diffusion controlled heterogeneous cure reaction. The low rate of diffusion in the resin beyond its gel point is controlled by high monomer rigidity, low mobility and radius of gyration of the monomers. The unreacted C=C bonds exist in the form of pendant, side groups chemically attached to the network. In addition, a hypothesis has been proposed that for concentrations of rigid bis-GMA monomers above 50 wt %, preferably only one methacrylate group of its molecule has reacted on the time span of the experiment performed and has been incorporated in the cured network. Hence, a substantial part of unsaturation exists in the solid polymer in the form of pendant groups formed by mechanically ineffective bis-GMA monomers. The results are in good qualitative agreement with computer simulations based on the kinetic gelation model. PMID- 15348073 TI - Variation in flexural properties of photo-pultruded composite archwires: analyses of round and rectangular profiles. AB - Prototype continuous, unidirectional, fiber-reinforced composite archwires were manufactured into round and rectangular profiles utilizing a photo-pultrusion process. Both 0.022 inch (0.56 mm) diameter and 0.021 x 0.028 inch (0.53 x 0.71 mm) rectangular composites were formed utilizing commercially available S2-glass reinforcement within a polymeric matrix. Reinforcement was varied according to the number, denier and twists per inch (TPI) of four S2-glass yarns to volume levels of 32-74% for round and 41-61% for rectangular profiles. Cross-sectional geometry was evaluated via light microscopy to determine loading characteristics; whereas two flexural properties (the elastic moduli and flexural strengths) were determined by 3-point bending tests. Morphological evaluation of samples revealed that as the TPI increased from 1 to 8, the yarns were more separated from one another and distributed more peripherally within a profile. For round and rectangular profiles utilizing 1 TPI fibers, moduli increased with fiber content approaching theoretical values. For round profiles utilizing 1 TPI and 4 TPI fibers, flexural strengths increased until the loading geometry was optimized. In contrast, the flexural strengths of composites that were pultruded with 8 TPI fibers were not improved at any loading level. Doubling the denier of the yarn, without altering the loading, increased both the moduli and flexural strengths in rectangular samples; whereas, the increases observed in round samples were not statistically significant. At optimal loading the maximum mean moduli and strengths equaled 53.6 +/- 2.0 and 1.36 +/- 0.17 GPa for round wire and equaled 45.7 +/- 0.8 and 1.40 +/- 0.05 GPa for rectangular wires, respectively. These moduli were midway between that of martensitic NiTi (33.4 GPa) and beta-titanium (72.4 GPa), and produced about one-quarter the force of a stainless steel wire per unit of activation. Values of strengths placed this composite material in the range of published values for beta-titanium wires (1.3-1.5 GPa). PMID- 15348074 TI - Tensile properties and transformation temperatures of Pd added Ti-Ni alloy dental castings. AB - The effect of palladium (Pd) addition to Ti-Ni alloy as the third element was investigated to improve the super-elasticity of the alloy castings at body temperature for dental application. Ti-50.8Ni (at %) alloy, which exhibited super elasticity at 310 K in castings, was used for comparison. 5.0, 7.5, 10.0 and 15.0 at % Pd was added to Ti-50.0Ni alloy by the substitution for Ni. The change in the proportion of Ti and Ni was also examined at the fixed Pd addition of 7.5 at %. The properties of the alloys were investigated in tensile test and differential scanning calorimetry (DSC). Ti-42.5Ni-7.5Pd alloy castings showed good super-elasticity among the examined alloys from the viewpoint of residual strain and elongation. Moreover, apparent proof stress could be changeable by the proportion of Ti and Ni with residual strain being kept low. Ti-42.5Ni-7.5Pd alloy castings exhibited better super-elastic flexibility than Ti-50.8Ni alloy, which is proven by lower apparent proof stress and larger elongation. This flexibility appears to be caused by its relatively high martensitic transformation starting temperature point. It is suggested that this flexibility with super-elasticity could widen the clinical application of the alloy casting in dentistry. PMID- 15348075 TI - Effect of skin-penetrating enhancers on the thermophysical properties of cholesteryl oleyl carbonate embedded in a thermo-responsive membrane. AB - The effect of skin-penetrating enhancers such as propylene glycol (PG), Azone and ethanol on the thermophysical properties of cholesteryl oleyl carbonate (COC) was investigated using differential scanning colorimetry (DSC) and microscopic Fourier-transform infrared (FT-IR) spectroscopy. The results indicate that PG did not influence the DSC thermograms and IR spectra of COC in the different ratios of COC and PG mixture; whereas Azone interacted with COC not only to lower the semectic-cholesteric phase transition temperature of COC but also to induce a new IR spectral peak at 1653 cm(-1) which shifted from the carbonyl stretching band (1636 cm(-1)) of Azone. Ethanol did not interact with COC, but it influenced the IR spectral peak intensity of COC at 1253 cm(-1). The peak intensity at 1253 cm( 1) gradually rose with the time of ethanol evaporation and was similar to that of the temperature effect. The solubility parameter was also used to explain the miscibility and interaction between COC and PG, Azone or ethanol. PMID- 15348076 TI - Synthesis and characterization of functional gradient materials using Indian corals. AB - The partial conversion of coralline hydroxyapatite (HA) into tricalcium phosphate (TCP) catalyzed by the thermal decomposition of silver oxide was investigated. The thermal analysis of HA powder mixed with 5 mol % Ag2O indicated the decomposition of silver oxide to be around 550 degrees C in the presence of HA. Subsequently a functional gradient structure was formed by spreading silver oxide on one surface of the HA compact of diameter 10 mm and then firing at 700 degrees C in air. The X-ray powder diffraction analysis confirmed the formation of alpha TCP induced by the Ag2O decomposition. The content of alpha-TCP of the sintered pellet was found to decrease with the increasing depth from the surface of the pellet. In vitro solubility study in phosphate buffer of pH 7.2 showed the stability of the biphasic material as in between HA and TCP. PMID- 15348077 TI - Bone healing in porous implants: a histological and histometrical comparative study on sheep. AB - Tissue integration in four types of porous implant materials (Interpore 200 or Corallin hydroxyapatite, hydroxyapatite blocks, hydroxyapatite granules and polymethylmethacrylate) was evaluated in vivo. Porous blocks measuring 20 mm x 10 mm x 8 mm were implanted in mandibles and iliac crests of sheep. Bone healing in porous blocks was studied at 2 and 6 months after implantation. The behavior of the material itself was also analyzed. Histological and histomorphometrical analysis revealed bone healing depending upon healing time and material. On the basis of analysis of variance, differences in amounts of bone ingrowth at 2 and 6 months were statistically significant (p=0.0039 in mandible; p=0.0351 in iliac crest). The longer the time span, the more mineralized tissues were observed in the specimen. Our data confirmed that hydroxyapatite has osteoconductive capacities. Porous PMMA was found to be biocompatible, but it showed less bonegrowth within the pores. Interpore 200, which had the highest surface to volume ratio was found to display the highest level of osseointegration and biodegradation. PMID- 15348078 TI - Effect of sintering parameters on the density and microstructure of carbonate hydroxyapatite. AB - This work documents an investigation into the effect of water on the density and microstructure of carbonate hydroxyapatite in carbon dioxide sintering atmospheres. Carbonate apatites with carbonate contents of between 3.2 and 7.8 wt % were precipitated and the precipitates were formed into dry gels. Isothermal and isochronal sintering experiments were performed under dry carbon dioxide and wet carbon dioxide (containing 3 wt % water) atmospheres. The effect of carbonate content was studied by using two gels both with a green density of 37% and with carbonate contents of 5.8 and 7.8 wt %. Both isothermal and isochronal experiments demonstrated that bloating of the apatite occurred and this behavior was associated with the loss of carbonate from the apatite. It was found that only in wet carbon dioxide atmospheres fully dense translucent carbonate apatite could be formed. 93% dense carbonate apatite was formed after 4 h sintering at temperatures as low as 700 degrees C. PMID- 15348079 TI - The processing and characterization of animal-derived bone to yield materials with biomedical applications. Part II: milled bone powders, reprecipitated hydroxyapatite and the potential uses of these materials. AB - Further studies on the processing and use of animal-bone-derived calcium phosphate materials in biomedical applications are presented. Bone powders sourced either from the direct crushing and milling of bovine, ovine and cervine bone or after being subjected to defatting and acid digestion/NaOH reprecipitation and sodium hypochlorite hydrogen peroxide treatment of animal bones were characterized using Fourier transform infra-red (FTIR) spectroscopy, 13C solid state magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy, atomic absorption (AA) and inductively coupled plasma (ICP) spectrometric techniques. Bone powders were trialled for their potential use as a substrate for phosphine coupling and enzyme immobilization as well as a feedstock powder for plasma spraying on titanium metal substrates. Results indicated that enzyme immobilization by phosphine coupling could be successfully achieved on milled cervine bone with the immobilized enzyme retaining some activity. It was found that the presence of impurities normally carried down with the processing of the bone materials (viz., fat and collagen) played an important role in influencing the adsorbency and reactivity of the powders. Plasma spraying studies using reprecipitated bovine-derived powders produced highly adherent coatings on titanium metal, the composition of which was mostly hydroxyapatite (Ca10(PO4)6(OH)2) with low levels of alpha-tricalcium phosphate (alpha-Ca3(PO4)2) and tetracalcium phosphate (Ca4P2O9) also detected. In general, animal derived calcium phosphate materials constitute a potentially cheaper source of calcium phosphate materials for biomedical applications and make use of a largely under utilized resource from abattoir wastes. PMID- 15348080 TI - The processing and characterization of animal-derived bone to yield materials with biomedical applications. Part III: material and mechanical properties of fresh and processed bovine cancellous bone. AB - Conversion of bovine cancellous bone to a useful biomedical xenograft material involves several processing steps which include boiling, defatting and deproteination (i.e. bleaching). This study has shown how these processes can influence cancellous bone modulus and strength. It was found that prolonged boiling in water for six hours followed by NaOCl bleaching had a deleterious effect on the overall strength of the bovine bone. In contrast, bone samples subjected to only moderate boiling (1.5 hours) exhibited a 22% stiffness increase due mainly to the effects of drying. The same stiffened samples, when subjected to the bleaching procedure, retained some strength with only a small reduction in moduli values. It can be concluded that careful control of defatting and bleaching procedures on bovine bone is able to give a strong, albeit, brittle material with preservation of the original bone architecture. The bone xenograft materials are worthy of further investigation in in vivo clinical trials to assess their performance in contact with biological fluids. PMID- 15348081 TI - Microstructure of ceramic coating on titanium surface as a result of hydrothermal treatment. AB - Hydroxyapatite coating on commercially pure titanium has been produced by a biomimetic method in order to improve osteointegration for medical implant purposes. A specific chemical treatment by etching titanium substrate with different concentrations of NaOH aqueous solution at 130 degrees C in an autoclave, followed by heat treatment at 600 degrees C was selected to obtain an activated titanium substrate. The microporous surface obtained has allowed the nucleation and growth of a calcium phosphate layer by soaking the substrate in a simulated body fluid (SBF). Scanning electron microscopy (SEM) together with energy dispersive analyzer for X-ray (EDS), X-ray diffraction (XRD) as well as Fourier transform infrared spectroscopy (FT-IR) were employed to evaluate the hydroxyapatite coating. A homogeneous structure coating without cracks defined the chemical treatment condition of the substrate. PMID- 15348082 TI - Physical and mechanical behavior of electron-beam irradiated and ethylene oxide sterilized multiblock polyester. AB - The effect of two sterilization treatments (electron-beam radiation and ethylene oxide gas) on the structure and mechanical properties of a multiblock copolymer were investigated to establish the effects of the sterilizing procedures on potential biomedical material. The material was exposed for different radiation doses in order to find an optimum dose of electron-beam radiation. Characterization techniques employed include gel permeation chromatography, infrared spectroscopy, differential scanning calorimetry, dynamic mechanical thermal analysis and tensile testing. The optimal dose of radiation at which no change in structure and mechanical properties occurred was found as 25 kGy. Ethylene oxide gas treatment also did not affect the structure and properties of the polymer and it can be recommended as an alternative sterilization route for the studied polymer. PMID- 15348083 TI - Comparison of mechanical properties for equiaxed fine-grained and dendritic high palladium alloys. AB - Two Pd-Cu-Ga alloys and a Pd-Ga alloy were selected for study. Bars of each alloy were tested in tension for the as-cast and simulated porcelain-firing conditions, and values of mechanical properties were measured. Fracture surfaces and microstructures of axially sectioned fracture specimens were observed with the SEM. The two Pd-Cu-Ga alloys exhibited similar mechanical properties. The Pd-Ga alloy had lower strength and higher percentage elongation. Heat treatment simulating porcelain firing cycles decreased the strength of both Pd-Cu-Ga alloys and increased their ductility. However, this heat treatment did not significantly affect the mechanical properties of the Pd-Ga alloy. All three high-palladium alloys had the same modulus of elasticity. The amount of overall porosity was relatively minimal (< 1%) and not significantly different among the three alloys. However, porosity was a significant factor for UTS of one Pd-Cu-Ga alloy and the Pd-Ga alloy. PMID- 15348084 TI - Development of soluble glasses for biomedical use Part I: in vitro solubility measurement. AB - In this study soluble glasses have been developed for biomedical applications containing P2O5 as a network former and CaO and Na2O as modifiers. This study shows that as expected, the glasses have an inverse exponential relationship between solubility and CaO content. Furthermore, there is evidence for compositional related non-linearity in the dissolution of the glasses with time. This is thought to be due to either layer formation on the glass surface hindering ion diffusion, ion exchange process or change of ionic strength of the solution. Bioactivity of these glasses is indicated by the formation of a brushite precipitate, a precursor to apatite formation. Further evidence for bioactivity is also presented in the second part of this paper. PMID- 15348085 TI - Development of soluble glasses for biomedical use Part II: the biological response of human osteoblast cell lines to phosphate-based soluble glasses. AB - Soluble glasses are considered to be of potential clinical value in orthopaedic and dental surgery. However, the biological response to these materials is not well understood. To determine the effects of these glasses, two human osteoblast cell lines, MG63 and HOS (TE85), were incubated in vitro in the presence of increasing concentrations of extracts of the glasses. The effects of the extracts on cell growth was measured using the MTT assay and an ELISA assay was used to measure the expression of bone sialoprotein (BSP), osteonectin (ON) and fibronectin (FN), antigens which play a fundamental part in the integrity and function of hard connective tissue. The results showed that the proliferation of the cells was adversely affected only by the more soluble glasses, which also down-regulated the expression of the bone-associated proteins. In contrast, the extract of the glass with the lowest dissolution rate, which contains relatively elevated levels of Ca2+, was found to enhance bone cell growth and antigen expression. These findings suggest that the compositions of these glasses at least partly determine the response of cells and thus, that the glasses could be modified to elicit a more optimal biological response and clinical efficacy. PMID- 15348086 TI - Dynamic mechanical characterization of hydroxyapatite reinforced polyethylene: effect of particle size. AB - Dynamic mechanical analysis (DMA) was used to characterize biomedical composites consisting of synthetic hydroxyapatite (HA) particulate reinforced polyethylene (PE). The effects of the HA volume fraction, temperature and HA particle on the storage modulus (E(I)) and damping (tan delta) were investigated. Increasing HA volume fractions increased E(I) and decreased tan delta. E(I) was found to be linearly related to the Young's modulus values obtained from quasi-static tensile tests. Relative modulus and damping studies showed that the viscoelastic behavior of unfilled PE was different to that of the filled matrix due to the presence of thermally induced tensile stresses in the matrix at the filler-matrix interface. PMID- 15348087 TI - Bone fracture analysis on the short rod chevron-notch specimens using the X-ray computer micro-tomography. AB - Mechanical fatigue of bone leads to micro-cracking which is associated with remodeling, establishing a balance in the microcrack population of the living tissue, thus, in the steady-state, the microstructure of bone provides sites of discontinuity acting as stress raisers. Hence fracture toughness plays a decisive role in bone functionality by determining the level to which the material can be stressed in the presence of cracks, or, equivalently, the magnitude of cracking which can be tolerated at a given stress level. Cortical bone, which behaves as a quasi-brittle solid when fractured, was tested as short-rod chevron-notched tension specimens (CNT). The main features of the CNT specimen are its geometry and the V shaped notch. The notch leads to steady-state crack propagation whilst the requested geometry allows a diameter 40% smaller than the thickness of a standard compact tension specimens (CT). These features are essential to distinguish the inhomogeneties in the fracture properties of materials like bone. Bone structure and crack propagation of the CNT specimens were analyzed using X ray computed micro-tomography (XMT), which is a non-invasive imaging technique. The unique feature of the micro-CT is the high resolution three-dimensional image which consists of multi-sliced tomographs taken in a fine pitch along the rotational axis. Fracture toughness (K(IC)) computed according to the peak load was 4.8 MNm(-3/2) while that derived from experimental calibration tests using XMT was 4.9 MNm(-3/2). PMID- 15348088 TI - Effect of hydrophobicity on in vitro streptococcal adhesion to dental alloys. AB - Non-specific interactions such as electrostatic interactions, and surface free energy are of importance in bacterial adhesion to dental surfaces as they determine whether or not bacteria are attracted to the surface. The relationship between adherence of Streptococcus mitis, S. mutans, S. oralis and S. sanguinis on precious and non-precious dental alloys, and the bacterial and alloy surface hydrophobicities (a measure of the surface free energy) was studied. The number of adhering bacteria was determined by fluorescence microscopy counts. The hydrophobicity of the bacteria and alloy surfaces were evaluated by adhesion to hexadecane and water contact angles, respectively. Our results showed that (i) the surfaces of the tested alloys were hydrophobic, (ii) S. sanguinis, S. mutans and S. oralis were hydrophobic, and (iii) S. mitis was hydrophilic. S. oralis, the more hydrophobic strain, demonstrated the highest adherence on the tested materials, whereas S. mitis adhered least on the hydrophobic surfaces. For the tested alloys, bacterial adherence was highest for the high gold content alloy, and lowest for the non-precious alloy. Our results showed that for the tested bacterial strains, there was a significant correlation between bacterial adhesion and substratum hydrophobicity: hydrophobic metal surfaces favor adhesion of hydrophobic bacteria. PMID- 15348089 TI - Plain and drug loaded polyphosphazene membranes and microspheres in the treatment of rabbit bone defects. AB - The healing of periodontal surgical defects was studied in rabbits, using polyphosphazenes (POP) membranes and microspheres, both plain or drug-enriched. POP polymers having amino acid ester as backbone substituents, are used since they resorb and undergo hydrolytic degradation to ammonia, phosphate and amino acids. Fourteen animal were operated in tibia, and other fourteen at angle of the mandible, that was reached by extraoral access. Bone defects were performed in tibiae, and covered either with POP or with poly-tetrafluoroethylene (PTFE) membranes, while other rabbits served as controls. The animals were sacrificed after one and two months, and the tibiae taken and processed for optical microscopy. Similar surgical defects were made in mandible, and POP membranes were placed over the breaches, some of which were filled with POP microspheres, both alone or mixed with granular hydroxyapatite. For comparison, two rabbits were treated with PTFE membranes, while other two served as controls. The animals operated at the mandible were all sacrificed after one month, and the operated bones taken and processed for histology. It was found that POP membranes were very effective in promoting the healing in tibiae, while less satisfactory results were found in the animals treated with PTFE membranes and in controls. In mandible, the healing occurred without a clear relationship with the grafted microgranular material or the membrane, since repairing bone was found also in controls. In any case, both POP membranes and microspheres showed excellent biocompatibility, as no inflammatory cells or macrophages were found in the surrounding tissue. This property was completely independent from the presence of drug, since the matrix-entrapped drugs, released in the tissue, did not hamper the bone healing. It was also found that POP, by itself, has a positive effect in stimulating the bone repair. PMID- 15348090 TI - Adhesion and proliferation of keratinocytes on ion beam modified polyethylene. AB - Polyethylene (PE) foils were modified by irradiation with Ar+ and Xe+ ions to different fluences and different physico-chemical properties of the irradiated PE were studied in relation to adhesion and proliferation of keratinocytes on the modified surface. Changes in the PE surface roughness were examined using the AFM technique, the production of conjugated double bonds and oxidized structures by UV-VIS and FTIR techniques respectively. The surface polarity was determined by measuring surface contact angle and two-point technique was used for the determination of PE sheet resistance. Adhesion and proliferation of keratinocytes was characterized using the MTT-test. The ion irradiation leads to creation of conjugated double bonds which, together with progressive carbonization, contribute to the observed decrease of sheet resistance. Oxidation of the irradiated PE surface layer during the ion implantation is observed. Besides oxidation, the PE surface polarity is affected by other factors. The observed increase of the PE surface roughness due to the ion irradiation is inversely proportional to the ion size. The adhesion and proliferation of keratinocytes on the ion irradiated PE is significantly higher than on the pristine PE. Distribution of results in keratinocyte cultivation and the number of cells is related to the ion fluence applied and to ion species as well. PMID- 15348091 TI - On fatigue lifetimes and fatigue crack growth behavior of bone cement. AB - Bone cement is used to develop a mechanical bond between an artificial joint and the adjacent bone tissue, and any degradation of this bond is of serious concern since it can lead to loosening and eventually malfunction of the artificial joint. In the present study, the fatigue lives and fatigue crack propagation behavior of two bone cements, CMW Type 3 and Zimmer, were investigated, and it was found that the size and distribution of pores played a major role in influencing both the fatigue crack initiation and propagation processes. The fatigue lifetimes of CMW exceeded those of Zimmer because of a lesser density of large pores. When the fatigue lifetimes were plotted as a function of Klimax, the maximum initial stress intensity factor based upon the initiating pore size, the difference in fatigue lifetimes between CMW and Zimmer bone cements was greatly reduced. The fatigue crack growth behavior of both bone cements were similar. This is a further indication that the noted differences in fatigue lifetimes were related to the size of the pore at the crack initiating site. PMID- 15348092 TI - Influence of thermal treatments on Ag Sn Cu powders in order to reduce mercury contents in dental amalgam. AB - The mercury content of dental amalgams is a controversial subject with regard to the biological properties of these materials. The object of this study is to optimize the thermal treatments performed on an experimental powder in order to obtain a low mercury ratio (41% by weight) while preserving the desirable clinical qualities of the material. Using atomized powder, two types of thermal treatments are performed: A1, to obtain a partially annealed structure and A2, to obtain a complete homogenization. The kinetics of the amalgamation reaction is mainly evaluated by X-ray diffraction to identify the newly formed phases as a function of setting time. Mechanical properties are evaluated according to the ISO norms at 37 degrees C. Metallographical examination of the amalgams shows a Ag-Hg phase which acts as a matrix incorporating the Cu-Sn and Ag-Sn compounds. The setting kinetics of the A1 amalgams is linear and slightly more rapid than that of the A2 amalgam. The mechanical properties of the amalgams are significantly improved regarding the higher mercury content commercial amalgams. A specific thermal treatment permits us to slow down the diffusion of mercury between the different intermetallic compounds into the powder. The final amalgam composition, thus, most approaches the stoechiometric ratio calculated from a quaternary diagram. PMID- 15348093 TI - Needle-like apatite-leucite glass-ceramic as a base material for the veneering of metal restorations in dentistry. AB - A needle-like apatite-leucite glass-ceramic was prepared in the SiO2-Al2O3-Na2O K2O-P2O5-F system. Nucleation and crystallization processes were studied in bulk and powdered samples. The crystallization of leucite follows the mechanism of surface crystallization. After the precipitation of NaCaPO4 crystals and another unknown crystal phase, the formation of needle-like apatite is based on a volume nucleation and crystallization process. The mechanism of the formation of needle like apatite differs to those of apatite precipitation in glass-ceramics. The morphology of needle-like apatite is comparable to that of apatite in natural teeth. The properties of the glass-ceramic, especially the good chemical durability, the optical properties, as well as mechanical and thermal properties allow glass-ceramic to be used as a main component in a bio-material for the veneering of metal restorations in dentistry. PMID- 15348094 TI - Incorporation of amino acids within the surface reactive layers of bioactive glass in vitro: an XPS study. AB - Surface reaction layers grown on bioactive glass (Bioglass), by immersion in either simulated body fluid (SBF) or minimal essential medium (alpha-MEM) for 2, 5, 32 and 72 h, were analyzed by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Layers grown in alpha-MEM exhibited delamination when observed in SEM. Low resolution XPS analysis detected nitrogen at the surface of the Bioglass exposed to alpha-MEM for 72 h (8 relative at%), whereas insignificant nitrogen was found at the surface of any sample immersed in SBF. XPS depth profiling (argon) showed the presence of nitrogen throughout the depth of the surface layer of the sample incubated in alpha-MEM for 72 h. Deconvolution of the nitrogen envelope in a high resolution XPS spectrum demonstrated nitrogen characteristic of the amine bonds present in amino acids. Carbon concentration also considerably increased over time with exposure to alpha MEM (24-55%), whereas it remained in the 20-25% range in SBF. These results demonstrate that the amino acids contained in the culture medium were incorporated within the growing calcium phosphate rich surface reaction layer of Bioglass. PMID- 15348095 TI - Evaluation of the acute scarring response to the implant of different types of biomaterial in the abdominal wall. AB - Since the short-term, acute scarring process induced by a biomaterial may condition the evolution of the repair process, the present investigation evaluates the behavior of polytetrafluoroethylene (PTFE) and polypropylene (PL) biomaterials in the initial stages of repair. Three PTFE biomaterials (Mycro Mesh, Dual Mesh and Soft Tissue Patch) and one PL biomaterial (Marlex) were employed to repair defects created in the abdominal wall of New Zealand rabbits. Animals were sacrificed at 3 or 7 days. Specimens were obtained for light and scanning electron microscopy, and immunohistochemical analysis using the RAM-11 monoclonal antibody for rabbit macrophages. The PL implants showed substantial adhesion formation with viscera. Lower adhesion formation was detected in the PTFE implants. The evolution of the acute phase of the repair process was similar for each PTFE biomaterial. At 3 days post implant, an incipient neoperitoneum was detected which was fully established after 7 days. The behavior of the PL implant was similar, although a greater amount of reticular granulation was detected. The neoformed peritoneum was irregular. Few RAM-11-labeled macrophages were detected in all cases. The acute phase of the tissue repair process induced by the implant of PTFE and PL biomaterials generally proceeds along similar lines to a normal repair process. However, the use of microporous, laminar materials seems to favor the early establishment of a well-defined neoperitoneal layer. PMID- 15348096 TI - Structure of dextran-magnetite complex: relation between conformation of dextran chains covering core and its molecular weight. AB - A homologous series of dextran-magnetite complex (DM) was synthesized using alkali-treated dextrans with various molecular weights from 1800 to 27000. The structure of DM particles in water, particularly effect on molecular weight of dextran, was studied. The number of dextran chains binding to a core is possibly determined by the steric hindrance between dextran chains covering core. The conformation of dextran chains may change from a fully extended state to a random coiled state with an increase in molecular weight. PMID- 15348097 TI - New non-woven polyurethane-based biomaterials for the cultivation of hepatocytes: expression of differentiated functions. AB - A new non-woven polyetherurethane support suitable to host cultured hepatocytes has been developed. Prior to its use in bioreactors and artificial liver devices, the biocompatibility of this new material was investigated. The experiments have shown that the survival and functionality of hepatocytes entrapped in the non woven polymer were longer than that of monolayer cultured hepatocytes, under serum-free culture conditions. Hepatic specific metabolic functions, namely, synthesis of urea and synthesis and secretion of plasma proteins, were well maintained by hepatocytes entrapped in non-woven polyetherurethane sheets. Cells also retained the expression of biotransformation activities of 7-ethoxycoumarin O-deethylase as well as CYP2A1, CYP2B1 and CYP3A1. The results presented in this paper point to non-woven polyetherurethane sheets as a suitable biocompatible support for functional, three-dimensional hepatocyte cultures. PMID- 15348098 TI - Precipitation behavior of Ag-Pd-In dental alloys. AB - Age-hardening characteristics and precipitation behavior of Ag-25%Pd-3%In-1%Zn 0.5%Ir alloy were investigated in detail by means of hardness testing, X-ray diffraction, electron microscopy and resistivity measurement. The solution treating could be accomplished at 980 degrees C and the aging in the temperature range from 950 to 850 degrees C occurred by continuous precipitation. The aging in the temperature range from 850 to 450 degrees C occurred first, forming GP zones with a hardness increase and then in overaging stage by forming discontinuous precipitation, which consisted of lamellae of solute (Pd, In, Zn) depleted Ag-rich phase and (Pd,Ag)3(In,Zn) intermetallic phase. The hardness increased very fast to its peak in 10 min during aging at temperatures between 450 and 550 degrees C. PMID- 15348099 TI - Bending and fracture of compact circumferential and osteonal lamellar bone of the baboon tibia. AB - Lamellar bone is common among primates, either in the form of extended planar circumferential arrays, or as cylindrically shaped osteons. Osteonal bone generally replaces circumferential lamellar bone with time, and it is therefore of much interest to compare the mechanical properties and fracture behavior of these two forms of lamellar bone. This is, however, difficult as natural specimens of circumferential lamellar bone large enough for standard mechanical tests are not available. We found that as a result of treatment with large doses of alendronate, the lateral sides of the diaphyses of baboon tibia contained fairly extensive regions of circumferential lamellar bone, the structure of which appears to be indistinguishable from untreated lamellar bone. Three-point bending tests were used to determine the elastic and ultimate properties of almost pure circumferential lamellar bone and osteonal bone in four different orientations relative to the tibia long axis. After taking into account the differences in porosity and extent of mineralization of the two bone types, the flexural modulus, bending strength, fracture strain and nominal work-to-fracture properties were similar for the same orientations, with some exceptions. This implies that it is the lamellar structure itself that is mainly responsible for these mechanical properties. The fracture behavior and morphologies of the fracture surfaces varied significantly with orientation in both types of bone. This is related to the microstructure of lamellar bone. Osteonal bone exhibited quite different damage-related behavior during fracture as compared to circumferential lamellar bone. Following fracture the two halves of osteonal bone remained attached whereas in circumferential lamellar bone they separated. These differences could well provide significant adaptive advantages to osteonal bone function. PMID- 15348100 TI - Phase transformation behavior in a multipurpose dental casting gold alloy during continuous heating. AB - Phase transformation in a multipurpose dental casting gold alloy during continuous heating was studied by electrical resistivity measurements, hardness tests, X-ray diffraction and scanning and transmission electron microscopy. The behavior can be explained by the following reaction sequences in the nodule: alpha1(fcc) + alpha2(L1(2)) --> alpha1(fcc) + alpha2(L1(2)) + beta(L1(0)), where fcc is face centred cubic. A discontinuous precipitation with very fine nodules contributed to the hardening and the growth produced the softening. This multipurpose gold alloy is characterized by the introduction of a PtZn ordered phase with L1(0) structure instead of a CuAu I phase. PMID- 15348101 TI - TEM analysis of tweed structure in high-palladium dental alloys. AB - Transmission electron microscopy and convergent-beam electron diffraction were used to study three high-palladium dental alloys: an as-cast Pd-Ga alloy, a cast Pd-Cu-Ga alloy aged at room temperature for over 5 years, and a cast Pd-Cu-Ga alloy annealed at 1023 K for 2 h and slowly cooled to room temperature. Bands containing a tweed structure in the Pd-Ga alloy were 120degrees twins with (1 1 0 mirror planes. Within these bands the alloy had a face-centered tetragonal structure with a c/a ratio of 1.03. The aged Pd-Cu-Ga alloy contained a larger amount of tweed structure than the original as-cast alloy, along with randomly oriented bands and thin lath-like regions. The annealed Pd-Cu-Ga alloy contained rectangular particles of a secondary phase that may be Pd2Ga, Pd5Ga2 or Pd13Ga5. The microtwinned band structure relieves the strain energy arising from transformation of the f.c.c. palladium solid solution on cooling. PMID- 15348102 TI - Numerical study of porosity in titanium dental castings. AB - A commercial software package, MAGMASOFT (MAGMA Giessereitechnologie GmbH, Aachen, Germany), was used to study shrinkage and gas porosity in titanium dental castings. A geometrical model for two simplified tooth crowns connected by a connector bar was created. Both mold filling and solidification of this casting model were numerically simulated. Shrinkage porosity was quantitatively predicted by means of a built-in feeding criterion. The risk of gas pore formation was investigated using the numerical filling and solidification results. The results of the numerical simulations were compared with experiments, which were carried out on a centrifugal casting machine with an investment block mold. The block mold was made of SiO2 based slurry with a 1 mm thick Zr2 face coat to reduce metal-mold reactions. Both melting and casting were carried out under protective argon (40 kPa). The finished castings were sectioned and the shrinkage porosity determined. The experimentally determined shrinkage porosity coincided with the predicted numerical simulation results. No apparent gas porosity was found in these model castings. Several running and gating systems for the above model casting were numerically simulated. An optimized running and gating system design was then experimentally cast, which resulted in porosity-free castings. PMID- 15348103 TI - Blood interactions with plasticized poly(vinyl chloride): relevance of plasticizer selection. AB - An investigation has been made of blood interactions with plasticized poly(vinyl chloride) (PVC) biomaterials in tubular form, taking into account the influence on the blood response of the polymer, antithrombotic agent, blood condition and test procedure. In vitro and ex vivo procedures were used to achieve a comparison between PVC plasticized with di- (2-ethylhexyl)phthalate (DEHP) and with tri-(2 ethylhexyl)trimellitate (TEHTM). The blood response was monitored in terms of the measurement of fibrinogen adsorption capacity, thrombin-antithrombin III complex (TAT) and the complement component C3a. Surface characterization of the polymers was performed by X-ray photoelectron spectroscopy (XPS). The data obtained indicate that in comparison with DEHP-PVC, there is a higher reactivity for TEHTM PVC, which correlates with the plasticizer distribution at the polymer surface. PMID- 15348104 TI - Toughening characterization in alumina platelet-hydroxyapatite matrix composites. AB - Fracture toughness of Al2O3 platelet-reinforced hydroxyapatite (HAP) ceramics was investigated using the Vickers' indentation technique. The geometrical anisotropy of alumina platelets induces an anisotropic toughening. The efficiency of reinforcing mechanisms remains maximum for a crack propagating with an angular deviation inferior to 30 degrees around the direction perpendicular to alumina disc faces. This is assumed to result from a crack deflection mechanism which induces a favorable contribution of mode II failure. A small effect of hydroxyapatite grain size becomes noticeable in the direction parallel to alumina disc faces. The toughening depends on the size and volume content of alumina platelets. Large size platelets provoke a spontaneous microcracking of the HAP matrix which is detrimental to the mechanical reliability, whereas small platelets lead to a strong toughening. The results relate to the intensity of thermoelastic residual stresses within the matrix around alumina inclusions. PMID- 15348105 TI - Effect of passivation on the dissolution behavior of Ti6A14V and vacuum-brazed Ti6A14V in Hank's ethylene diamine tetra-acetic acid solution Part I Ion release. AB - This work aims to investigate the effects of three factors, namely: (1) two differently prepared materials (as-polished Ti6A14V and 2 h brazed Ti6A14V); (2) three different surface passivation treatments (34% nitric acid passivation, 400 degrees C heated in air, and aged in 100 degrees C de-ionized water); and (3) periods of immersion time (up to 32 days), on trace element release in Hank's ethylene diamine tetra-acetic acid (EDTA) solution. After passivation and autoclaving treatment, the specimens were immersed in 8.0 mM EDTA in Hank's solution and maintained at 37 degrees C for periods of time up to 32 days. The 400 degrees C -treated specimens exhibit a substantial reduction in constituent release, which may be attributed to the higher thickness and rutile structure of the surface oxides. For acid-passivated and water-aged treatments, a highly significant decrease in the trace levels of Ti, A1, and V is detected from the brazed Ti6A14V compared to those obtained from the Ti6A14V specimens. It is hypothesized that an anatase-rutile transformation of surface TiO_2 is likely to occur, accelerated by the elements of copper and nickel in the brazed specimens. In addition, a significant time-related decrease in constituent release rate is observed for all kinds of specimens throughout the 0-8 day experimental period. The implication of the results is discussed. PMID- 15348106 TI - Bridging defects in nerve continuity: influence of variations in synthetic fiber composition. AB - Synthetic filaments introduced into a silicone tube may help to enhance axonal growth over extended defects in nerve continuity [1]. Here we test the influence of number (0, 3, 7 or 15), size (diameter 150 or 250 microm) and material of filaments (polyamide or catgut) enclosed in such tubes (inner diameter 1.98 mm) on axonal growth across a 10 mm defect in rat sciatic nerve. The morphology of the tube content was analyzed four weeks post-surgery. The area of the formed tissue matrix inside the tube showed no difference between the groups. Myelinated axons were observed in the formed tissue matrix inbetween and peripheral to the filaments, however, separated from the filaments by concentric cell layers. The number of myelinated axons was less in the tubes with 15 filaments, most pronounced when catgut filaments were used. In most cases, except in tubes with 15 catgut filaments, fibers had grown into the distal nerve segment (pinch reflex test/light microscopy). We conclude that an intrinsic framework consisting of a limited number of synthetic filaments inside an extrinsic framework (silicone tube) does not disturb nerve regeneration. The formed tissue matrix was neither influenced by the presence or the numbers (if less than or equal to seven filaments), type of filaments nor the size of the filaments indicating the importance of the inserted nerve segments. PMID- 15348107 TI - Magnetic properties of Ni-Co-Cr-base Elgiloy. AB - The Ni-Co-Cr-base Elgiloy is one of the commonly used engineering materials. Most applications rely on its high strength, ductility, corrosion resistance and excellent fatigue life over a wide temperature range. However, in the medical application of cerebral aneurysm clips, the alloy is often subjected to strong magnetic fields associated with magnetic resonance imaging (MRI). Its paramagnetic behavior meets MRI safety requirements, but is the source of relatively large artifacts and thus less MRI-compatible for MRI procedure involving the brain. This article reports superconducting quantum interference device (SQUID) measurements on the magnetic properties of a series of Elgiloy wires in either as-drawn or heat-treated conditions. Furthermore, low-temperature calorimetry was employed to reveal the existence of submicroscopic clusters containing ferromagnetic elements such as Ni or Co in the macroscopically paramagnetic matrix. PMID- 15348108 TI - Crystal growth of carbonate apatite using a CaCO3 flux. AB - Single crystals of carbonate apatite were grown using a CaCO3 flux under an Ar gas pressure of 55 MPa. The crystals obtained were observed by scanning electron microscopy, optical microscopy and X-ray diffraction. Electron probe microanalyses and thermal analyses were performed. CO3 ions in planar triangle form replaced both OH sites and PO4 tetrahedral sites in the apatite structure: in particular, the OH sites were perfectly substituted by CO3 ions using this method. PMID- 15348109 TI - In vitro growth and differentiation of osteoblast-like human bone marrow cells on glass reinforced hydroxyapatite plasma-sprayed coatings. AB - Human osteoblastic bone marrow cells were cultured for periods of up to 28 days in control conditions and on the surface of a glass reinforced hydroxyapatite composite (HA/G1) and commercial hydroxyapatite (HA) plasma-sprayed coatings, in the "as-received" condition and after immersion treatment in culture medium for 21 days. Cultures were characterized for total protein content and alkaline phosphatase activity. Scanning electron microscope analyses were performed on control cultures, seeded materials and materials incubated in the absence of cells. Culture media were analyzed for total and ionized calcium and phosphorus concentrations throughout the incubation period. Immersion of HA/G1 and HA coatings in culture medium resulted in significant alterations to the levels of calcium and phosphorus in the medium, leading to surface modifications. However, seeded material samples showed significant differences in the pattern of variation of the levels of these species. Cell proliferation was observed in the "as-received" HA/G1 composite, but cell mediated formation of mineral deposits was not proved. In contrast, "as-received" HA hardly supported cell growth. Previously immersed material samples showed cell proliferation and evidence of biological formation of mineral deposits. However, the HA/G1 composite presented better surface characteristics for cell growth as the behavior of bone marrow cells was closer to that observed in control cultures. PMID- 15348110 TI - The long-term interaction of dental cements with lactic acid solutions. AB - A study of the interaction of dental cements with lactic acid solutions has been carried out in which individual cement specimens were repeatedly exposed to 20 mmol dm(-3) lactic acid for periods of a week. After each week of storage, the mass of the specimens was recorded and the pH of the solution determined. The glass-ionomers showed an initial increase in mass, followed by a decline that became steady from 6 weeks. Zinc polycarboxylate and zinc phosphate cements, by contrast, showed no early gain in mass, but eroded steadily more or less from the start of their exposure to lactic acid. For all cements, acid erosion followed linear kinetics, at rates ranging from 0.5%/week for the zinc phosphate to 0.28%/week for one of the glass-ionomers, Chelonfil (ESPE, Germany). At the end of six months, the zinc phosphate had lost 14.2% of its initial mass, the zinc polycarboxylate 9.9% and the glass-ionomers between 6.2 and 7.2%. Erosion was accompanied on every occasion by neutralization of the acid solution. Both erosion and neutralization continued steadily throughout the experiment. The effectiveness of neutralization was in the following order: zinc polycarboxylate>zinc phosphate>glass-ionomer. The pH change in Week 1 was much greater for the glass-ionomers and the zinc polycarboxylate than in all subsequent weeks. PMID- 15348111 TI - Characterization of anodic spark-converted titanium surfaces for biomedical applications. AB - The aim of the present study was to characterize the surface morphology, microstructure and the chemical composition of anodic spark-converted titanium surfaces. The coatings were prepared in an electrochemical cell by the anodic spark deposition technique in an aqueous solution of Ca(H2PO4)2. The coatings were characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), electron probe microanalysis (EPMA) and X-ray diffraction (XRD). The properties of the coatings are described in terms of morphology. PMID- 15348112 TI - Preparation of phosphorus-containing silica glass microspheres for radiotherapy of cancer by ion implantation. AB - A chemically durable glass microsphere containing a large amount of phosphorus is useful for in situ irradiation of cancers, since they can be activated to be a beta-emitter with a half-life of 14.3 d by neutron bombardment. When the activated microspheres are injected to the tumors, they can irradiate the tumors directly with beta-rays without irradiating neighboring normal tissues. In the present study, P+ ion was implanted into silica glass microspheres of 25 microm in average diameter at 50 keV with nominal doses of 2.5 x 10(16) and 3.35 x 10(1)6 cm(-2). The glass microspheres were put into a stainless container and the container was continuously shaken during the ion implantation so that P+ ion was implanted into them uniformly. The implanted phosphorus was localized in deep regions of the glass microsphere with the maximum concentration at about 50 nm depth without distributing up to the surface even for a nominal dose of 3.35 x 10(16) cm(-2). Both samples released phosphorus and silicon into water at 95 degrees C for 7 d. On the basis of the previous study on P+-implanted silica glass plates, the silica glass microspheres containing more phosphorus which is desired for actual treatment could be obtained, without losing high chemical durability, if P+ ion would be implanted at higher energy than 50 keV to be localized in deeper region. PMID- 15348113 TI - Bioactive nanocrystalline sol-gel hydroxyapatite coatings. AB - Sol-gel technology offers an alternative technique for producing bioactive surfaces for improved bone attachment. Previous work indicated that monophasic hydroxyapatite coatings were difficult to produce. In the present work hydroxyapatite was synthesized using the sol-gel technique with alkoxide precursors and the solution was allowed to age up to seven days prior to coating. It was found that, similar to the wet-chemical method of hydroxyapatite powder synthesis, an aging time is required to produce a pure hydroxyapatite phase. A methodology that has been successfully used to produce nanocrystalline hydroxyapatite thin film coatings via the sol-gel route on various substrates including alumina, Vycor glass, partially stabilized zirconia, Ti-6Al-4V alloy and single crystal MgO is described. Coatings produced on MgO substrates were characterized by X-ray diffraction and atomic force microscopy, while the analogous gels were examined with thermogravimetric and differential thermal analyses. The coatings were crack free and the surface was covered with small grains, of approximately 200 nm in size for samples fired to 1000 degrees C. Coating thickness varied between 70 and 1000 nm depending on the number of applied layers. PMID- 15348114 TI - The effect of strontium oxide in glass-ionomer cements. AB - The reaction of strontium oxide powder with poly(acrylic acid) has been studied both alone and within glass-ionomer cements. Reaction was found to be slow and the strontium-carboxylate structure was found to be partially covalent in character, as determined by Fourier transform infrared spectroscopy (FTIR). These are similar to the structures formed by calcium in glass-ionomer cements, but are different from typical monomeric strontium carboxylates, which tend to be purely ionic. Strontium oxide powder introduced in two types of glass-ionomer cements, slowed down the setting reaction at both 21 degrees C and 37 degrees C, but at low levels (5 wt %), increased the compressive strength in both cement formulations studied. However, at higher levels, it was found to decrease the compressive strength. This study confirms the view that strontium is a cement forming ion; but concludes that, except at very low levels, strontium oxide powder does not improve the properties of glass-ionomer cements. PMID- 15348115 TI - Study of diopside ceramics for biomaterials. AB - Diopside was prepared by sintering a powder compact of composition CaO-MgO-2SiO2 at 1300 degreesC for 2 h. The bending strength of diopside was 300 MPa and the fracture toughness was 3.5 MPa m1/2. It was proved that diopside has no general toxicity in cell culture. Diopside implanted in rabbits came in close contact with the newly grown bone. X-ray microanalysis spectral diagrams show a change of composition across the junction from the diopside to the newly grown bone. High resolution transmission electron microscopy revealed crystal growth at the interface between diopside and the newly grown bone, and continuity between diopside lattices and those of the new crystals. PMID- 15348116 TI - Characterization of SiO2-Na2O-Fe2O3-CaO-P2O5-B2O3 glass ceramics. AB - Bioactivity and magnetic properties were investigated in glass and glass ceramics based on the SiO2-Na2O-Fe2O3-CaO-P2O5-B2O3 system to find their suitability as thermoseed for hyperthermia treatment of cancer. The effect of change in compositions on bioactivity was examined in simulated body fluids. The glass ceramic samples exhibit Na3CaSi3O8 and Na3-XFeXPO4 phases. After dipping the glass ceramic samples in simulated body fluids silica hydrogel first forms, followed by an amorphous calcium phosphate layer. Magnetic and microwave resonance experiments further demonstrate the potential of these glass ceramics for possible use in hyperthermia. PMID- 15348117 TI - Effects of drying on the mechanical properties of bovine femur measured by nanoindentation. AB - Effects of drying on the measurement of mechanical properties of bone by nanoindentation methods have been examined. Tests were conducted to measure the elastic modulus and hardness of two cross-sectional cortical specimens obtained from adjacent areas of bovine femur. One specimen was thoroughly dried in air prior to testing while the other was stored in deionized water. The properties of osteons and interstitial lamellae showed statistically significant differences (p<0.0001) and were therefore investigated separately. Drying was found to increase the elastic modulus by 9.7% for interstitial lamellae and 15.4% for osteons. The hardness was also found to increase by 12.2% for interstitial lamellae and 17.6% for osteons. PMID- 15348118 TI - Effect of tannic acid solution on collagen structures for dental restoration. AB - This study examined the effect of tannic acid solution on dissolution of dentine collagen and morphological aspects of tendon collagen. Using root dentine, which was cut off from bovine anterior tooth, dentine powders were obtained by the pulverization and lyophilization. They were subject to an application of 1, 3, 5 or 10% tannic acid (TA) solution for 1, 3, 6, 12 or 24 h. TA-treated dentine powders were treated with 40% phosphoric acid (PA) for 30 s at 20 degrees C and additionally with trypsin. Released hydroxyproline in Woessner's assay after a hydrolysis in 6 N HCl at 110 degrees C for 20 h was assumed to be dissolved dentine collagen. Released hydroxyproline in a control sample without acid treatment decreased from 100 to about 60% with increased TA concentration of 1 to 10%, and decreased with increased incubation times of 1 to 24 h when applied by 5% TA solution. Scanning electron microscopy results established the morphological effect of their surface characteristics due to such treatments as 40% PA for 30 s and 5% TA for 6 h, or 40% PA after 5% TA treatment, yielding collagen structures protected by TA to attack from phosphoric acid. PMID- 15348119 TI - Effect of convergence angle and luting agent on the fracture strength of In ceram crowns. AB - This study compared the fracture strength of In ceram crown shapes fabricated with either 8 or 16 degrees total occlusal convergence; and attached with either a commercial zinc phosphate cement or a glass ionomer cement. Thirty crown shapes (8 mm diameter and 8.5 mm high) were fabricated for each preparation design on a brass master die with approximately the same dimensions as a premolar. In ceram crown shapes were luted on to the die using zinc phosphate or glass ionomer. The crown shapes were fractured in a testing machine (Instron) using a steel ball, 4 mm in diameter, that contacted the occlusal surface and the resulting data were statistically analyzed using a Mann-Whitney test. The results indicate that there is no statistical difference in the fracture strength values between preparations with 8 degrees total convergence compared with 16 degrees using the same cement. However, crown shapes luted with zinc phosphate on preparations with 8 and 16 degrees total occlusal convergence were significantly stronger than those luted using glass ionomer cements (p<0.05). PMID- 15348120 TI - The influence of the time-dependent properties of bone cement on stress in the femoral cement mantle of total hip arthroplasty. AB - An empirically determined formula for the creep behavior of bone cement was incorporated into a validated computer model of a cemented femoral total hip arthroplasty component. The stress patterns in the cement mantle were observed over a period of one week, in one instance where the stem-cement interface was rigidly bonded, and in a second, where it was allowed to slip. Principal stresses and maximum shear stresses were shown to decrease rapidly after loading in both situations, suggesting that the stresses generated were not high enough to cause immediate failure, although they may be significant in the long term. PMID- 15348121 TI - The effect of processing temperature and time on the structure and fracture characteristics of self-reinforced composite poly(methyl methacrylate). AB - A novel material, self-reinforced composite poly(methyl methacrylate) (SRC-PMMA) has been previously developed in this laboratory. It consists of high-strength PMMA fibers embedded in a matrix of PMMA derived from the fibers. As a composite material, uniaxial SRC-PMMA has been shown to have greatly improved flexural, tensile, fracture toughness and fatigue properties when compared to unreinforced PMMA. Previous work examined one empirically defined processing condition. This work systematically examines the effect of processing time and temperature on the thermal properties, fracture toughness and fracture morphology of SRC-PMMA produced by a hot compaction method. Differential scanning calorimetry (DSC) shows that composites containing high amounts of retained molecular orientation exhibit both endothermic and exothermic peaks which depend on processing times and temperatures. An exothermic release of energy just above Tg is related to the release of retained molecular orientation in the composites. This release of energy decreases linearly with increasing processing temperature or time for the range investigated. Fracture toughness results show a maximum fracture toughness of 3.18 MPa m1/2 for samples processed for 65 min at 128 degrees C. Optimal structure and fracture toughness are obtained in composites which have maximum interfiber bonding and minimal loss of molecular orientation. Composite fracture mechanisms are highly dependent on processing. Low processing times and temperatures result in more interfiber/matrix fracture, while higher processing times and temperatures result in higher ductility and more transfiber fracture. Excessive processing times result in brittle failure. PMID- 15348122 TI - Development of porous spherical hydroxyapatite granules: application towards protein delivery. AB - A new method for the preparation of porous spherical hydroxyapatite granules is reported. It may be clinically applied towards orthopaedic or maxillofacial surgery as fillers or packing materials, and as biological chromatography supports. Its application towards delivery of macromolecules or protein drugs is discussed utilizing human serum albumin (HSA) as a model protein. PMID- 15348123 TI - The influence of surface condition on the localized corrosion of 316L stainless steel orthopaedic implants. AB - The localized corrosion of austenitic stainless steel 316L intended for use as orthopaedic implants is determined as a function of the surface condition and metallurgical state. From the examination of samples exposed to a ferric chloride solution, at both 22 and 37 degrees C, the independent contribution of crevice and pitting corrosion to localized corrosion is determined. Both forms of localized corrosion occur to a greater extent at the higher temperature. The results indicate that weight loss measurements may not be sufficient to determine the extent of crevice corrosion separately from the influence of pitting corrosion. More importantly, the surface conditions required for the best resistance to crevice or pitting corrosion differ. Electropolished surfaces provide the best resistance to crevice corrosion, while "bead blasted" surfaces provide the best resistance to pitting corrosion. The implication of this result in terms of the serviceability as orthopaedic implants is discussed. The current results indicate the cold-worked state exhibits improved resistance to pitting corrosion. However, the influence of the metallurgical state could not be separated from a possible compositional effect. PMID- 15348124 TI - In-vitro calcium phosphate growth over functionalized cotton fibers. AB - Biomimetic growth of calcium phosphate compound on cotton sheets treated with tetraethoxy silane and soaked in simulated body fluid solution was studied using scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), micro-Fourier transform infrared spectroscopy (FTIR) and X-ray diffractometry (XRD). Micro-FTIR and EDAX results show that silicon was coupled to the cotton fiber when cotton was treated with tetra-ethoxy silane (TEOS) at 125 degrees C for 1 h. Calcium phosphate nucleation started to occur on the surface of TEOS treated cotton fibers upon immersion in 1.5xSBF (simulated body fluid solution) within 3 days and after 20 days, all the fiber surfaces were found covered with a thick and porous coating of calcium phosphate. The Ca and P determined by inductively coupled plasma spectroscopy (ICP) analysis revealed that the Ca/P ratio as well as the amount of calcium phosphate coating depends on the soaking time in SBF solution. PMID- 15348125 TI - Interfacial bond strength of electrophoretically deposited hydroxyapatite coatings on metals. AB - Hydroxyapatite (HAp) coatings were deposited onto substrates of metal biomaterials (Ti, Ti6Al4V, and 316L stainless steel) by electrophoretic deposition (EPD). Only ultra-high surface area HAp powder, prepared by the metathesis method 10Ca(NO3)2 + 6(NH4)2HPO4 + 8NH4OH), could produce dense coatings when sintered at 875-1000degreesC. Single EPD coatings cracked during sintering owing to the 15-18% sintering shrinkage, but the HAp did not decompose. The use of dual coatings (coat, sinter, coat, sinter) resolved the cracking problem. Scanning electron microscopy/energy dispersive spectroscopy (SEM/EDS) inspection revealed that the second coating filled in the "valleys" in the cracks of the first coating. The interfacial shear strength of the dual coatings was found, by ASTM F1044-87, to be approximately 12 MPa on a titanium substrate and approximately 22 MPa on 316L stainless steel, comparing quite favorably with the 34 MPa benchmark (the shear strength of bovine cortical bone was found to be 34 MPa). Stainless steel gave the better result since -316L (20.5 microm mK(-1)) > alpha-HAp (approximately 14 microm mK(-1)), resulting in residual compressive stresses in the coating, whereas alpha-titanium (approximately 10.3 microm mK( 1)) < alpha-HAp, resulting in residual tensile stresses in the coating. PMID- 15348126 TI - Diaphyseal fractures treated by polylactide and hydroxyapatite pins. Experimental study in rat. AB - In order to develop a biodegradable interlocking nail for fracture fixation, polylactic acid (PLA) pins and hydoxyapatite pins were implanted in the femoral bone in rats. A distal fracture was performed. The union and the tissue reaction to PLA and hydroxyapatite versus stainless steel rods were studied after 15 days, 1, 2 and 6 months implantation. Metal and PLA pins induced a union. Hydroxyapatite pins (Ossatite) did not prevent callus formation, but did not lead to consolidation in all cases because of weakness of the gelatin matrix binding the apatite particles together. PLA and stainless steel pins induced the same union and a similar tissue reaction during the studied implantation of 6 months. The biocompatility of Ossatite is satisfactory and the osteo-inductive properties of hydroxyapatite was confirmed. With injectable Ossatite, we could not obtain rat femoral fracture consolidation. We can confirm good biomaterial tolerance in bone which contrasts with important soft tissue reactions. Use of such material should be carefully limited to filling intra-osseous cavities. PMID- 15348127 TI - Electrochemical field effects in biological materials: electro-osmotic dewatering of cancerous tissue as the mechanistic proposal for the electrochemical treatment of tumors. AB - Some recent excellent papers on the electrochemical treatment (ECT) of tumors provide observations for an analysis of this phenomenon from an electrochemical point of view. An attempt is made here to develop the idea that ECT is a case of electro-osmotic dewatering (EOD) of the tumor material with the consequent changes in pH, with the concomitant role of reactions at the electrodes. Some quantitative considerations explaining the role of electrochemical double layers, zeta potential and associated quantities during the electro-osmotic process are outlined. A capsule summary of the factors that may be involved in necrosis of the tumor tissue is given in which the electro-osmotic dewatering at the anode and excess water accumulation at the cathode are the paramount effects, together with the associated pH changes. PMID- 15348128 TI - Brain lesion-induced alteration of selected phenotypic properties of spleen macrophages and their partial restoration in the course of foreign body reaction against intraperitoneally implanted polymers. AB - A lesion in the dorsoposterior part of the rat brain septum is known to exert an inhibitory effect on the delayed skin hypersensitivity and incorporation of radiolabeled thymidine into the lymphoid organs. To determine whether distinct properties of macrophages will also be modulated by this type of injury, we have focused upon the monitoring of expression of sugar receptors (lectins). In this study we show a reduction in the number of macrophages expressing carbohydrate binding sites for asialoglycoproteins (beta-D-galactoside), alpha-D-mannoside and alpha-D-mannoside-6-phosphate in spleen macrophages after the lesion of the dorsoposterior septum of the brain in the rat. The number of ED-1+ macrophages was not influenced. The intraperitoneal injection of beads prepared from the copolymer of 2-hydroxyethyl methacrylate with dimethyl aminoethyl methacrylate (30 wt %) elevated significantly the number of ED-1+ spleen macrophages and number of macrophages with binding site(s) recognizing asialoglycoproteins and alpha-D-mannoside-6-phosphate, respectively. These results indicate that a foreign-body reaction appears to be able to mediate a phenotypic restoration of lectin expression by spleen macrophages altered by the brain lesion. It can be suggested that, for example, a probable production of cytokines by the inflammatory cells colonizing the implanted beads plays a role in this process. PMID- 15348129 TI - Effect of chitosan and dextran on the properties of poly(vinyl alcohol) hydrogels. AB - Hydrogels are three-dimensional polymeric networks very similar to biological tissues and potentially useful as drug delivery systems. Poly(vinyl alcohol) based hydrogels containing different amounts of dextran or chitosan were prepared using the freezing-thawing method. Repeated freezing-thawing cycles of a poly(vinyl alcohol) (PVA) aqueous solution lead to the formation of crystallites which act as cross-linking sites, and a hydrogel with a high capacity to swell is obtained. The effects of the two different polysaccharides on the properties of the obtained materials were investigated by differential scanning calorimetry, dynamic mechanical analysis and scanning electron microscopy. In addition the release with time of poly(vinyl alcohol) in aqueous medium, was monitored and evaluated. On the basis of the obtained results it seems that the presence of dextran favors the crystallization process of PVA, allowing the formation of a more ordered and homogeneous structure. Instead, chitosan seems to perturb the formation of PVA crystallites leading to a material with a less regular structure. PMID- 15348130 TI - Evaluation by Vickers indentation of fracture toughness of a phosphate biodegradable glass. AB - Indentation tests are commonly used for the evaluation of fracture toughness of brittle materials, particularly glasses and ceramics, because this technique requires only a small polished area on the specimen surface from which a large number of data points can be generated rapidly. However, a wide variety of equations for the calculation of fracture toughness of ceramic materials by means of Vickers indentation are available. Such equations are obtained phenomenologically and their parameters adjusted in such a way that the KIC values obtained are in good agreement with those obtained by conventional methods. This is the reason why it is necessary to check which type of equation reproduces more accurately the results obtained by means of conventional methods for the material which is going to be investigated. In the present work seven different fracture toughness equations widely used in glass and ceramic studies are considered and the results are compared with those obtained by conventional methods, such as single-edge notch beam (SENB) specimens tested in three-point bending. The role played by the applied indentation load is considered. PMID- 15348131 TI - Improvement of haemocompatibility of metallic stents by polymer coating. AB - An alternative to open heart surgery in treating arterial diseases causing restricted blood flow is the implantation of intracoronary metallic stents. In spite of the advances in implantation and in spite of the excellent mechanical properties of metallic stents, there are still limitations because of the thrombogenicity of the metal. We have, hence, directed our attention to the coating of metallic stents with an ultrathin polymer layer by chemical vapor deposition (CVD) polymerization of 2-chloroparacyclophan. In a second step of surface modification the poly(2-chloroparaxylylene) layer is modified by treatment with a sulfur dioxide plasma in order to obtain a more hydrophilic surface with new functional groups. The results demonstrate the stable polymer coating of the stents and the improvement of haemocompatibility after treatment with sulfur dioxide plasma. Platelet adhesion is decreased from 85% for the metal surface to 20% for the CVD-coated and sulfur-dioxide-plasma treated surface. PMID- 15348132 TI - The failure of amalgam dental restorations due to cyclic fatigue crack growth. AB - In this study a restored mandibular molar with different Class II amalgam preparations was examined to analyze the potential for restoration failure attributed to cyclic fatigue crack growth. A finite element analysis was used to determine the stress distribution along the cavo-surface margin which results from occlusal loading of each restoration. The cyclic crack growth rate of sub surface flaws located along the dentinal cavo-surface margin were determined utilizing the Paris law. Based on similarities in material properties and lack of fatigue property data for dental biomaterials, the cyclic fatigue crack growth parameters for engineering ceramics were used to approximate the crack growth behavior. It was found that flaws located within the dentine along the buccal and lingual margins can significantly reduce the fatigue life of restored teeth. Sub surface cracks as short as 25 microm were found capable of promoting tooth fracture well within 25 years from the time of restoration. Furthermore, cracks longer than 100 microm reduced the fatigue life to less than 5 years. Consequently, sub-surface cracks introduced during cavity preparation with conventional dental burrs may serve as a principal source for premature restoration failure. PMID- 15348133 TI - No biological advantage with a low temperature curing versus a conventional bone cement: an experimental, mechanical and histomorphometrical study in the rabbit tibia. AB - Both tibial marrow cavities of 12 rabbits were evacuated and filled with curing bone cement. In one of the tibias conventional curing bone cement (Simplex P) was injected, while the other tibia of the same animal was filled with a low temperature curing bone cement (Boneloc). Three titanium implants were inserted along the proximal metaphysis of each tibia. Eight weeks after insertion the most distal implant in each tibia was removed while recording the removal torque. The implant was then once again screwed home into its bone bed. The animals were sacrificed 16 weeks after implant insertion. The previously removed implant and another implant in each tibia were then both removed while recording the removal torque. The third implant in each tibia was cut out en bloc with surrounding tissue and processed for ground section. We found no statistical differences in the mechanical or the histomorphometric evaluation of implant integration between the two cements, indicating that the low temperature curing bone cement does not result in a significantly different bone response from that of a conventional acrylic cement. PMID- 15348134 TI - Comparative wear under four different tribological conditions of acetylene enhanced cross-linked ultra high molecular weight polyethylene. AB - In this study, the wear of ultra high molecular weight polyethylene (UHMWPE) (Grade RCH 1000) crosslinked by gamma irradiation in acetylene was compared to virgin (non-irradiated) UHMWPE using four different wear configurations: (i) unidirectional motion with a smooth counterface, (ii) multidirectional motion with a smooth counterface, (iii) unidirectional motion with a rough counterface and (iv) multidirectional motion with a rough counterface. PMID- 15348135 TI - Development of thin elastomeric composite membranes for biomedical applications. AB - A breakthrough has been made in blending of two immiscible biocompatible polymers to form thin transparent interpenetrating network composite membranes (CM) with exceptional improvement in properties. Two immiscible polymers, namely the biaxially drawn ultra high molecular weight polyethylene (UHMWPE) film and polyether polyurethane (PU) were used. The fabrication included solution casting and heat compaction. During the fabrication, the CM still preserved the orientation of UHMWPE fibers but introduced the interpenetration of PU in UHMWPE film. The intimate interaction of PU with UHMWPE fibers was viewed through the transparency of CM. Differential scanning calorimetry (DSC) data showed the melting temperature (Tm) of UHMWPE increased by about 10 degrees C in CM and about 5 degrees C in heat-compacted membranes (HCM). Morphological observations indicated that CM presented a layered structure while HCM was a dense material without obvious void inclusions. The ultimate tensile strength and relative Young's modulus of CM are about 62 MPa and 460 MPa, respectively. They are about four times greater in strength and 150 times greater in modulus compared with those of PU. Heat compaction resulted in a membrane with nearly five times the tensile strength and 50 times the Young's modulus of PU. The engineered ultimate strain of CM is about 26%, 8% more than that of the porous UHMWPE film while about 70% of HCM, which is a 50% increase achieved through heat compaction. The tensile fracture toughness is about 93 mJ for CM and 211 mJ for HCM, two and five times that for the porous UHMWPE film, respectively. The significant modification on the properties of the heat-compacted composite may raise broad interest in using the CM to develop membrane-related devices and organ covers in biomedical applications. PMID- 15348136 TI - Quantitative analysis of the wear and wear debris from low and high carbon content cobalt chrome alloys used in metal on metal total hip replacements. AB - The biological reactions to polyethylene wear debris have been shown to result in osteolysis and loosening of total hip arthroplasties. This has led to renewed interest in the use of metal on metal bearings in hip prostheses. This study employed uniaxial and biaxial multistation pin on plate reciprocators to assess how the carbon content of the cobalt chrome alloy and the types of motion affected the wear performance of the bearing surfaces and the morphology of the wear debris generated. The low carbon specimens demonstrated higher wear factors than both the mixed carbon pairings and the high carbon pairings. The biaxial motion decreased the wear rates of all specimens. Plate wear was significantly reduced by the biaxial motion, compared to pin wear. The metal wear particles isolated were an order of magnitude smaller than polyethylene particles, at 60-90 nm, and consequently, 100-fold more particles were produced per unit volume of wear compared to polyethylene. The low carbon specimens produced significantly larger particles than the other material combinations, although it is thought unlikely that the difference would be biologically significant in vivo. The volumetric wear rates were affected by the carbon content of the cobalt chrome alloy, the material combination used and type of motion applied. However, particle morphology was not affected by the carbon content of the alloy or the type of motion applied. PMID- 15348137 TI - Functionalized copolymers and their composites with polylactide and hydroxyapatite. AB - Synthetic copolymers poly(epsilon-caprolactone-co-vinylphosphonic acid) (P(MDOVPA) and poly(epsilon-caprolactone-co-dimethylvinylphosphoester) (P(MDOVPE)) were used to prepare composites with polylactide (PLac) and hydroxyapatite (HAp). The P(MDOVPA) is used as filler in PLac films, as it has pendant functional groups P(O)(OH)2, providing nucleation sites for the deposition of HAp in simulated body fluid. HAp growth on P(MDOVPA) powder and PLac-P(MDOVPA) film was observed by Fourier transform infrared spectroscopy (FTIR), energy dispersive X-ray analysis (EDX) and X-ray diffraction (XRD). P(MDOVPE) and PLac blend are miscible, and the incorporation of hydrophilic P(MDOVPA) into PLac increased the hydrophilicity of the blend. Synthetic HAp was used to make multilayered, alternating organic-inorganic composites with porous PLac-P(MDOVPE) blends. PMID- 15348138 TI - Solid state spatially resolved 1H and 19F nuclear magnetic resonance spectroscopy of dental materials by stray-field imaging. AB - As part of a program to evaluate the use of stray-field magnetic resonance microimaging (STRAFI) in dental materials research spatially resolved nuclear magnetic resonance (NMR) for solid dental cements has been investigated. By applying a quadrature echo pulse sequence to a specimen positioned in the stray field of a NMR spectrometer superconducting magnet the magnetic resonance within a thin slice was obtained. The specimen was stepped through the field in 500 microm increments to record 1H and 19F profiles and T2 values at each point. The specimens were fully cured cylinders made from four types of restorative material (glass ionomer, resin modified glass ionomer, compomer, composite). The values for 1H T2 varied with material type and reflected the nature of the matrix structure. For all materials containing 19F in the glass two values were calculated for 19F T2, one short and one long. These were relatively invariant. Solid state magic angle spinning (MAS)-NMR showed that they came from the glass. This suggests that a proportion of the element is relatively mobile (in a glass phase) and the remainder is more tightly bound (in a compound dispersed in the glass). This demonstration, that NMR microimaging of both 1H and 19F in solid dental cements is possible, opens up exciting new possibilities for investigating the distribution of these elements (in particular fluorine) in solid dental materials. PMID- 15348139 TI - Effect of filler content and size to properties of composite resins on microwave curing. AB - The effect of filler content and size to property of composite resin using high purity spherical silica particles on microwave curing was examined by mechanical means and electron microscope observation to develop microwave-curing composite resin inlay. Increasing filler content in three kinds of filler particles (0.45, 0.96 and 1.46 microm) resulted in increasing compressive strength, diametral tensile strength and knoop hardness and indicated the highest values at the operative mixing limit. The effect of particle size was that the mechanical property of the 0.96 microm filled resin was highest, followed by 1.46 microm and 0.45 microm on the same filler content. The 0.96 microm filled resin had about the same knoop hardness on both surfaces of the cured sample, which means that uniform polymerization occurred in the cured sample. With electron microscope observation, the bubbles of about 1.0 microm and the cracks between the fillers and the matrix were observed in each cured resin. This is caused by the excessive absorption of microwave energy in fillers and surface treatment materials of fillers. Therefore, improvement of the composition of filler and surface treatment material of filler are needed, which makes it harder to absorb the microwave energy. PMID- 15348140 TI - Real-time measurement of in vitro enamel demineralization in the vicinity of the restoration-tooth interface. AB - An X-ray attenuation method using photon counting (scanning microradiography) is presented for the real-time study of in vitro demineralization of dental tissues in the vicinity of the restoration-tooth interface. By repeated measurement of mineral content profiles during the course of demineralization, the pattern of lesion development and the rate of mineral loss can be studied. The method is illustrated by comparison of enamel demineralization near a polyacid-modified composite resin restoration, near a bis-GMA/TEGMA composite resin restoration, and in an unrestored control. The method has potential for study of the influence of restorative materials on susceptibility of tooth tissue to demineralization. PMID- 15348141 TI - Microstructured bioreactive surfaces: covalent immobilization of proteins on Au(1 1 1)/silicon via aminoreactive alkanethiolate self-assembled monolayers. AB - Micrometer-scale patterns of a defined surface chemistry and structure were produced on both ultraflat Au(1 1 1) and on gold-coated monocrystalline silicon surfaces by a method combining microcontact printing, wet chemical etching and the replacement of etch-resist self-assembled monolayers (SAMs) by functionalized or reactive SAMs. Key steps in this methodology were characterized by X-ray photoelectron spectroscopy (XPS), ellipsometry and contact angle measurements. The covalent immobilization of (functional) biological systems on these surfaces was tested using an N-hydroxysuccinimide ester omega-functionalized disulphide (DSU), which covalently binds primary amines without the need for further activation steps. Atomic force microscope images of native collagen V single molecules immobilized on these patterned surfaces revealed both high spatial resolution and strong attachment to the monolayer/gold surface. Microcontact printing of DSU is shown to be feasible on specially prepared, ultraflat Au(1 1 1) surfaces providing a valuable tool for scanning probe experiments with biomolecules. The retention of enzymatic activity upon immobilization of protein was demonstrated for the case of horseradish peroxidase. The described approach can thus be used to confine biological activity to predetermined sites on microstructured gold/silicon devices - an important capability in biomedical and biomolecular research. PMID- 15348142 TI - Response to polyetherimide based composite materials implanted in muscle and in bone. AB - The in-vivo response to a composite material obtained with polyetherimide (PEI) reinforced with carbon/glass fibers was investigated by histological methods by implanting cylinders in muscle and in bone of the New Zealand White rabbit. A common metallic alloy, widely used in orthopaedic surgery, was used as control (Stellite). The aim of the study was to analyze the biological response towards the surface of the material. Composite implants and metallic implants did not induce adverse or inflammatory reactions. The morphological picture produced was similar, in muscle and in bone, for both materials. In muscle, cylinders were confined by an extremely thin fibrous layer and the overall appearance of the muscular tissue was normal. In bone, cylinders were confined by a nearly annular rim of newly formed bone. From these data it is possible to derive that the response to PEI-based composite material is comparable with the response to metallic substrate and, then, the material can be suitable for clinical application. PMID- 15348143 TI - Preparation and characterization of bioactive monolayer and functionally graded coatings. AB - Hydroxyapatite powders were made by reacting orthophosphoric acid with calcium hydroxide and dense bioactive coatings were subsequently produced by the plasma spray technique. Three types of hydroxyapatite (flame spheroidized) monolayer coatings and three types of functionally graded coatings were manufactured. It was found that average microhardness values of monolayer coatings decreased as the indentation load increased. The relationship between indentation load and indent diagonal length observed Meyer's law. Microhardness and fracture toughness of coatings were affected by characteristics of feedstock powders for plasma spraying. The indentation fracture toughness of coatings could be significantly increased by incorporating a toughening phase. PMID- 15348144 TI - Corrosion and biocompatibility of orthodontic wires. AB - With the increasing number of orthodontic treatments using devices containing nickel and the growing prevalence of nickel allergy in the average population, biocompatibility studies of these devices have become a topic of major interest. The corrosion behavior of orthodontic wires is a decisive factor determining their biocompatibility. Therefore four nickel-titanium guiding arches, a titanium molybdenum and a stainless steel wire were analyzed for corrosion behavior under realistic conditions. Pure potentiostatic, pure mechanical and combined potentiostatic and mechanical stresses were applied to the specimens. Subsequently, the surfaces of the wires were investigated employing atomic force microscopy (AFM) and nickel loss was measured with an atomic absorption spectrophotometer. The results yield information about the relative corrosion tendency of the wires under in vitro conditions. The wires examined can be classified into two groups, one with a high and a second group with a low tendency towards corrosion, that is American Orthodontics Memory wire as well as GAC Neo Sentalloy and Ormco Ni-Ti as well as Unitek Nitinol respectively. Although corrosion behavior under clinical conditions can not be directly derived from these results, analyses of wires after clinical usage indicate that changes of wire surfaces might show the same characteristics under in vitro conditions. PMID- 15348145 TI - New composites of hydroxyapatite and bioresorbable macromolecular material. AB - Composite materials were prepared by mixing in different proportions of hydroxyapatite (HA) and poly(epsilon-caprolactone-oxyethylene-epsilon caprolactone) block copolymer (PCL-POE-PCL) to produce a new resorbable material for biomedical applications. This material has proved to be very interesting for production of periodontal membranes. Mechanical properties are linearly proportional to the amount of HA introduced. Fourier transform infrared (FTIR) investigations have pointed out that HA is able to influence some close epsilon caprolactone molecules to start its homopolymerization giving PCL with an end chain ionic bonding. HA grains are therefore surrounded by a film of PCL which grants close connection of HA grains within copolymeric matrix. This interface bond with PCL is, however, an interesting occurrence for preparations of HA/PCL composites. PMID- 15348146 TI - Hardness of model dental composites - the effect of filler volume fraction and silanation. AB - The relationship between structure and mechanical properties for dental composites has often proved difficult to determine due to the use of commercially available materials having a number of differences in composition i.e. different type of resin, different type of filler, etc. This makes a scientific study of any one variable such as filler content difficult if not impossible. In the current study it was the aim to test the hypothesis that hardness measurements of dental composites could be used to monitor the status of the resin-filler interface and to determine the efficacy of any particle silanation process. Ten model composites formulated from a single batch of resin and containing a common type of glass filler were formulated to contain varying amounts of filler. Some materials contained silanated filler, others contained unsilanated filler. Specimens were prepared and stored in water and hardness (Vickers') was determined at 24 h using loads of 50, 100, 200 and 300 g. Composites containing silanated fillers were significantly harder than materials containing unsilanated fillers. For unsilanated products hardness was independent of applied load and in this respect they behaved like homogeneous materials. For composites containing silanated fillers there was a marked increase in measured hardness as applied load was increased. This suggests that the hardness-load profile could be used to monitor the status of the resin-filler interface. PMID- 15348147 TI - PVC as a sensor membrane material: influence of solvent casting variables. AB - Unplasticized polyvinyl chloride (PVC) has proved an especially difficult membrane material to form reproducibly. In its fabrication for sensor use, variable temperature solvent casting has been assessed and related to function as external membranes of a classical dual membrane amperometric oxidase electrode. The thermal history of the casting solution was shown to have an effect on both thickness and the permeability. With increasing temperature of the cast solution (4-37 degrees C) membrane thicknesses increased (5-30 microm). Amperometric responses to catechol and hydrogen peroxide decreased with increasing casting solution temperature whereas responses to ascorbate remained unchanged. These properties are tentatively attributed to the formation of discrete layers through differential changes in the rate of solvent evaporation. Scanning electron microscopy revealed distinct microstructure zones supporting this proposition and attesting to the need to factor in environmental temperature during casting. PMID- 15348148 TI - Hydrogels based on chitosan and dextran as potential drug delivery systems. AB - The release of human growth hormone (GH) from bioartificial polymeric materials in the form of hydrogels, was measured in vitro for up to 3 weeks. Poly(vinyl alcohol) (PVA) was blended, in different ratios, with two biological polymers, dextran and chitosan respectively. These blends were used to prepare hydrogels, using a freeze-thawing method. The hydrogels were loaded with GH, and their potential use as delivery systems was investigated. The release with time of PVA, in aqueous medium, was also monitored and evaluated. Scanning electron microscopy was used to investigate the structure of the hydrogels. The results obtained indicated that GH can be released from both dextran/PVA and chitosan/PVA hydrogels. The initial GH concentration used for sample loading affected the total quantity of GH released but not the pattern of release. The amount of GH released was affected by the content of the biological component. The percentage of PVA released was low but it was, however, related to the content of chitosan and dextran in the blends. PMID- 15348149 TI - Fracture evaluation of acrylic bone cements modified with hydroxyapatite: influence of the storage conditions. AB - Fracture and mechanical characterization of bone composite composed of polymethylmethacrylate and hydroxyapatite (HA) at different contents was carried out. Hydroxyapatite is added in order to improve cement biocompatibility, but it is expected that it also affects mechanical properties. Specimens were either stored in air at 37 degrees C for 120 h or in physiological solution (PhS-37), in order to establish the influence of storage conditions upon mechanical behavior. One set of specimens was also postcured at 120 degrees C for 4 h to take into account the influence of free monomer. Fracture experiments revealed some non linearity in load-displacement records and differences in trends between initiation and propagation values of the fracture surface energies. The trends in the data shows that HA acts as a rigid filler enhancing fracture resistance, flexural modules and yield stress, up to a certain content. Beyond the latter limit, properties suffer a deterioration because the addition of HA also affects the cement porosity. Absorbed water acts as plasticizer leading to a decrease in mechanical properties. The highest propagation strain energies were exhibited by materials aged in PhS-37. PMID- 15348150 TI - The implant material, Ti6Al7Nb: surface microstructure, composition and properties. AB - The excellent biocompatibility of titanium and its alloys is intimately related with the properties of the surface in contact with the biological environment, and therefore it is closely connected with the stable, passivating oxide layer that forms on its surface. In the present paper, the oxide layer on the alloy Ti6Al7Nb has been characterized using X-ray photoelectron spectroscopy, scanning Auger microscopy and pH-dependent lateral force microscopy. The alloying elements Al and Nb are incorporated in the oxide layer and detected in their most stable oxidized form, as Al2O3 and Nb2O5. Their distribution in the oxide reflects the underlying alpha-beta microstructure, with enrichment of Al in the alpha- and of Nb in the beta-phase (determined by electron microprobe). Friction measurements (lateral force microscopy) indicate slightly different, pH-dependent, lateral forces above the alpha- and beta-phase structures that point to small local variations in surface charges. PMID- 15348151 TI - Elaboration conditions influence physicochemical properties and in vivo bioactivity of macroporous biphasic calcium phosphate ceramics. AB - Two different preparations of biphasic calcium phosphate (BCP) were characterized in vitro: BCP1 from a mechanical mixture of hydroxyapatite (HA) and beta tricalcium phosphate (beta-TCP) powders, and BCP2 from calcination of a calcium deficient apatite (CDA). The structural, physicochemical and mechanical parameters of these two preparations were investigated, and two different macroporous BCP1 (MBCP1) and BCP2 MBCP2) implants were manufactured and implanted in rabbit bone for in vivo bioactivity studies. Scanning electron microscopy observations showed that MBCP1 implants had a significantly higher degradation rate (P<0.0001) than MBCP2 implants. This was probably caused by the presence of calcium oxide impurities in BCP1 and the more intimate mixture and stable ultrastructure of BCP2. No significant difference about the newly formed bone rate in these two BCP preparations was observed. Very slight variations in sintering conditions appeared to influence the biodegradation behavior of the two MBCP implants despite their identical HA/-TCP ratios and similar porosity. Precise and complete in vitro characterization enabled us to understand and predict in vivo degradation behavior. PMID- 15348152 TI - The formation of hydroxyapatite-calcium polyacrylate composites. AB - Tetracalcium phosphate (TetCP, Ca4(PO4)2O) reacts rapidly with polyacrylic acid (PAA). Complete reaction results in the formation of hydroxyapatite (HAp) and calcium polyacrylate. Consequently, this combination of reactants can react to form a dental cement. However, reaction occurs so rapidly that it would be difficult to achieve a homogeneous mixture of reactants suitable for use in restorations. In order to explore extending the working time, the effects of prehydrating the TetCP to form surface layers of HAp on the TetCP particles was explored. Prehydration was found to be an effective means of allowing workability. Therefore, the effects of the proportions of TetCP and PAA, with and without HAp filler, on cement properties were investigated. The extents of the reactions were investigated by X-ray diffraction analysis; the extents of PAA neutralization were studied by Fourier transform infra-red spectroscopy (FTIR); pore structures were determined by mercury intrusion porosimetry; microstructures were observed by scanning microscopy, and compressive strengths were determined. After curing for 17 days at room temperature PAA neutralization was almost complete; however, residual TetCP could be detected by X-ray diffraction and PAA by FTIR. As expected, the compressive strengths of the cements showed a dependence on the liquid (water+polymer)-to-solid (TetCP+HAp filler) used. The presence of HAp filler caused a significant decrease in compressive strength and increasing the proportion of HAp filler resulted in a decrease in the compressive strength. The characteristics of the load-deflection curves showed a dependence on the presence of HAp filler. In the absence of filler, two slopes were observed in the curves whereas a linear curve, typical of a ceramic, was observed when HAp filler was present. Mercury intrusion porosimetry (MIP) indicated the majority of the porosity was present in pores larger than 0.1 microm. Porosity increased with increasing liquid-to-solids ratio and with an increasing proportion of HAp filler at a constant liquid-to-solids ratio. Microstructural observations indicated the effect of HAp filler on increasing porosity was the result of porosity present in the filler itself. Thus, poorly consolidated HAp filler contributed to increased porosity and reduced compressive strength. PMID- 15348153 TI - Incorporation of a controlled-release glass into a calcium phosphate cement. AB - A so-called controlled-release glass was synthesized occurring in the system CaO Na2O-P2O5. A certain sieve fraction of this glass was incorporated in a calcium phosphate cement, of which the powder contained alpha-tricalcium phosphate (alpha TCP), dicalcium phosphate (DCP) and precipitated hydroxyapatite (HA). The glass appeared to retard the cement setting slightly and it reduced considerably the compressive strength after aging in aqueous solutions which were continuously refreshed. Scanning electron microscope (SEM) pictures and X-ray diffraction (XRD) patterns of the samples after 5 weeks of aging showed that the glass was not dissolved but that large brushite crystals were formed. Thereby, aging in CaCl2 solutions resulted in more brushite formation than aging in NaCl solutions. The brushite crystals did not reinforce the cement. Neither was the aged glass containing cement weaker than it was before the brushite formation right after complete setting. In conclusion, the incorporation of controlled-release glasses into a calcium phosphate cement and subsequent aging in aqueous solutions did not result in the formation of macropores in the cement structure, but that of brushite crystals. This incorporation reduced the compressive strength of the cement considerably. PMID- 15348154 TI - Compositional and structural control in bone regenerative coatings. AB - The development of a low-temperature method of producing bioactive coatings for medical implants has been shown to bypass the problems associated with high temperature processing routes, in particular the appearance of amorphous phases and non-stoichiometric hydroxyapatite (HA), and delamination of the coating from the substrate. An electric field/aqueous solution technique for producing adherent, crack-free calcium phosphate coatings on titanium and stainless steel substrates is described. The characteristics of the coating are a function of electrode spacing, supersaturation, temperature and current and voltage conditions. Scanning electron microscopy (SEM) characterized the surface morphology of the coatings, which were shown to be HA. The possibility of producing a coating of carbonate-substituted HA having the same chemical composition as bone apatite, and forming at physiological temperatures, has also been demonstrated. The size of the microstructure decreased and the morphology changed as the carbonate ion concentration in the calcium and phosphate ion solution increased. PMID- 15348155 TI - Production and characterization of new calcium phosphate bone cements in the CaHPO4-alpha-Ca3(PO4)2 system: pH, workability and setting times. AB - The initial setting properties of calcium phosphate cements in the CaHPO4-alpha Ca3(PO4)2 (DCP-alpha-TCP) system have been investigated. Interest was focused on the pH, workability, cohesion time and initial and final setting times. The addition of CaCO3 modified the structure of the cement reaction product such that it became more similar to the apatite phase in bone mineral. The addition of 10% w/w of CaCO3 reduced the viscosity of the cement pastes resulting in an increase in initial and final setting times and improved injectability. PMID- 15348156 TI - Heterogeneous crystallization of dicalcium phosphate dihydrate on titanium surfaces. AB - The kinetics of nucleation and crystal growth of dicalcium phosphate dihydrate (DCPD, CaHPO4.2H2O) on titanium powders with different grain sizes have been investigated using the constant composition crystal growth method at 37 degrees C and pH = 5.50. Nucleation is independent of titanium powder size while crystal growth rate is strongly size-dependent. A minimum relative supersaturation ratio, sigma(min), required in the system for a detectable crystal growth rate to occur, taking into account the presence of the foreign substrate in the system, was determined. Based on the kinetic data and on the various characterizations by X ray photoelectron spectroscopy and by scanning electron microscopy, we propose a crystal growth kinetic equation as a function of two parameters: the size of the substrate and the excess of supersaturation (sigma-sigma(min)) which is squared, indicating a spiral growth mechanism. PMID- 15348157 TI - Porosity control of hydroxyapatite implants. AB - Conformation of hydroxyapatite (HAp) bodies was carried out using a multiple slip casting technique, in order to obtain dual-layer samples with differential sized porosities. The external layer, because of its porosity, controlled by the addition of organic compounds (polyvinyl polyacrylate, PVC), will promote bone ingrowth. The internal denser layer, due to the addition of lithium phosphate (Li3PO4) as sintering additive, will give mechanical resistance to the implant. HAp aqueous suspensions were characterized by rheological measurements. Scanning electron microscopy (SEM) and intrusion mercury porosimetry (MP) were used to characterize sintered bodies. It can be concluded that it is possible to introduce gross porosity in HAp bodies by the addition of organic compounds. The results show that a compatible shrinking of the layers during the sintering process and a good frequency of pores with an appropriate size in the external layer can be achieved with the use of organic additives. PMID- 15348158 TI - Morphology and composition of hydroxyapatite coatings prepared by hydrothermal treatment on electrodeposited brushite coatings. AB - Highly pure brushite (CaHPO4.2H2O) coatings on porous Ti6Al4V substrates were prepared by electrodeposition from aqueous electrolytes. The influence of hydrothermal treatment parameters on brushite-to-hydroxyapatite conversion and the morphology and phase composition of hydroxyapatite (HAP) coatings was studied. It was found that the content, Ca/P atomic ratio, grain size and pore size of HAP in coatings increase with increasing hydrothermal treatment temperature, and that increasing the pH value can promote brushite-to-HAP conversion and reduce the grain size of HAP. Under optimal conditions, highly pure HAP coatings with needle-like crystals and non-stoichiometric form, which are similar to those of calcium phosphate in human bone, can be obtained. PMID- 15348159 TI - Local drug delivery system using ceramics: vacuum method for impregnating a chemotherapeutic agent into a porous hydroxyapatite block. AB - We performed an experimental study on a new drug delivery system that employs a porous hydroxyapatite block (HAb) (composition: Ca10(PO4)6(OH)2)) to conduct sustained release of a chemotherapeutic agent. To confirm penetration of the agent into the HAb (2 cm3), an aqueous solution containing eosin dye was used at various vacuum pressures. To estimate the storage capacity of the HAb, blocks were weighed before and after being impregnated with the aqueous dye solutions, and the capacity of the block was calculated from the increase in weight after vacuum. In this slow-release study using vacuum, the anti-cancer drug methotrexate (MTX) was used in vitro. Four HAb (1 cm3) containing different concentrations of MTX, ranging from 1.22 to 2.38 mg per block, were studied. All were found to release the drug, maintaining a mean concentration of 0.22 to 0.32 microg/ml even after twelve days. This concentration is high enough to be effective against tumor cells. The results suggest that HAb impregnated with a chemotherapeutic agent using a simple vacuum system may serve as a valuable new method of administering local chemotherapy, primarily when used as a strut graft for bone defects. This new drug delivery system can also be used as an adjuvant material in extended curettage, which can also discourage recurrence of benign tumors without any risk of systemic toxicity. PMID- 15348160 TI - Synthetic collagen fibers coated with a synthetic peptide containing the YIGSR sequence of laminin to promote peripheral nerve regeneration in vivo. AB - The usefulness of collagen fibers and the YIGSR sequence (Tyr-lle-Gly-Ser-Arg) of laminin for nerve regeneration were examined in vivo. Type I collagen gel (G group), Type I collagen fibers (F-group), Type I collagen fibers coated with laminin (L-group) or the YIGSR sequence (Y-group) were packed into silicone tubes, 15 mm long, and transplanted to the sciatic nerves of Wistar rats. Empty silicone tubes were used as the control. The animals were sacrificed 8 weeks after transplantation. Bridging of the nerve was confirmed in the F-(7/12), Y (7/10) and L-group (6/10), but no bridging was observed in any of the animals of the G- and control group. Nerve regeneration among the space of collagen fibers was observed, and it was suggested that fibroblasts infiltrated the gap in the substance of the degenerated collagen fibers were followed by Schwann cells on the basis of immunocytochemistry. The number of myelinated axons per regenerated tissue in the tube (density), and total area of myelinated axons per measured regenerated tissue in the tube (% axon area) in each the L- and Y-group were significantly higher than that in the F-group (P < 0.05). These results suggest the possibility of obtaining adequate nerve regeneration with new artificial materials only. PMID- 15348161 TI - Characterization of porous hydroxyapatite. AB - Hydroxyapatite has been considered for use in the repair of osseous defects for the last 20 years. Recent developments have led to interest in the potential of porous hydroxyapatite as a synthetic bone graft. However, despite considerable activity in this field, regarding assessment of the biological response to such materials, the basic materials characterization is often inadequate. This paper documents the characterization of the chemical composition, mechanical integrity, macro- and microstructure of a porous hydroxyapatite, Endobon (E. Merck GmbH), intended for the bone-graft market. Specimens possesed a range of apparent densities from 0.35 to 1.44 g cm(-3). Chemical analysis demonstrated that the natural apatite precursor of Endobon was not converted to pure hydroxyapatite, but retained many of the ionic substituents found in bone mineral, notably carbonate, sodium and magnesium ions. Investigation of the microstructure illustrated that the struts of the material were not fully dense, but had retained some traces of the network of osteocyte lacunae. Macrostructural analysis demonstrated the complex inter-relationship between the structural features of an open pore structure. Both pore size and connectivity were found to be inversely dependent on apparent density. Furthermore, measurement of pore aspect ratio and orientation demonstrated a relationship between apparent density and the degree of macrostructural anisotropy within the specimens, while, it was also noted that pore connectivity was sensitive to anisotropy. Compression testing demonstrated the effect of apparent density and macrostructural anisotropy on the mechanical properties. An increase in apparent density from 0.38 to 1.25 g cm(-3) resulted in increases in ultimate compressive stress and compressive modulus of 1 to 11 MPa and 0.2 to 3.1 GPa, respectively. Furthermore, anisotropic high density (> 0.9 g cm(-3)) specimens were found to possess lower compressive moduli than isotropic specimens with equivalent apparent densities. These results underline the importance of full structural and mechanical characterization of porous ceramic implant materials. PMID- 15348162 TI - Water sorption behaviour of polymeric systems based on tetrahydrofurfuryl methacrylate. AB - Earlier research has described the water absorption behaviour, drug release and biological properties of a room temperature polymerizing system based on poly(ethyl methacrylate) (PEM) powder and tetrahydrofurfuryl methacrylate (THFM) monomer. This work has been extended, with respect to water sorption behaviour, by replacing the monomer to various extents with hydroxyethyl methacrylate (HEMA), and poly(ethyl methacrylate) by ethyl methacrylate (EM)-THFM copolymers. Replacing the THFM with HEMA, and gelling with PEM, increased the diffusion coefficient progressively. The replacement of PEM by EM-THFM copolymers, when gelled with THFM monomer, substantially reduced equilibrium water uptake, and increased diffusion coefficients. However, with HEMA monomer, equilibrium uptake was unaffected, but the diffusion coefficient decreased with increasing THFM content of the copolymer. This is due to a complex interaction of THFM cross linking the copolymer, and the effect of EM on the water uptake. Heat polymerizing the PEM-THFM system reduced equilibrium uptake and the diffusion coefficient, compared with the room temperature polymerizing system; this could reflect molecular weight differences. PMID- 15348163 TI - In vitro crystallization of octacalcium phosphate on type I collagen: influence of serum albumin. AB - The heterogeneous crystallization of octacalcium phosphate (OCP, Ca8H2(PO4)6.5H2O) on demineralized Type I collagen has been studied from metastable supersaturated solutions, at 37degreesC and pH=6.50, using the constant composition crystal growth technique. The induction period, before OCP crystal growth, varied markedly with the degree of supersaturation of the solution. The data obtained allowed us to determine the apparent order for the precipitation and the growth mechanism of OCP on Type I collagen. Infrared spectroscopy analyses indicated the progressive mineralization of collagen and observations by scanning electron microscopy confirmed the development of OCP crystals on the collagen surface. The influence of bovine serum albumin on both the kinetics of OCP nucleation and growth has also been investigated. Because this protein was adsorbed on calcium phosphate nuclei, it exhibited two distinct effects as a function of its concentration in solution. We proposed a mechanism explaining the interaction between albumin and calcium phosphate nuclei or crystals and its incidence on the OCP crystallization kinetics. Observations by scanning electron microscopy revealed a modification of the size and the appearance of crystals grown on collagen due to the adsorption of albumin on the crystal surface. PMID- 15348164 TI - Thin films of calcium phosphate and titanium dioxide by a sol-gel route: a new method for coating medical implants. AB - Titanium is a commonly used biomaterial for dental and orthopaedic applications. To increase its ability to bond with bone, some attempts were made to coat its surface with calcium phosphate (CaP). This paper describes a new type of coating. Instead of a pure CaP layer, a mixing of titanium dioxide (TIO2) and CaP is fabricated and deposited as a coating. These layers are deposited by a sol-gel route on pure titanium substrates using various pre-treatments. The method consists of mixing a solution of tetrabutyl ortho-titanate or a sol of titanium dioxide with a solution of calcium nitrate and phosphorous esters. This composite is deposited on to commercially pure titanium plates, mechanically polished or blasted with pure crystalline aluminum oxide, using the spin-coating technique. These coatings are then fired at 650 or 850 degrees C for various times. The samples are characterized by X-ray diffraction for their crystallinity, X-ray photoelectron spectroscopy for their surface chemical composition and scanning electron microscopy for their topography. Samples treated at 850 degrees C present a well-pronounced crystallinity, and a high chemical purity at the surface. The topography is strongly related to the viscosity of the precursor and the substrate pre-treatment. Possibilities to structure the outermost layer are presented. PMID- 15348165 TI - Calcium phosphate bone cements for clinical applications. Part I: solution chemistry. AB - Calcium phosphate cements have been the subject of many studies in the last decade because of their biocompatibility, their capacity to fill bone cavities and their hardening properties; properties which are desirable in a broad range of surgical applications. The setting and hardening of these materials are controlled by dissolution-precipitation chemical reactions at room or body temperature and involve crystalline phase transformations. PMID- 15348166 TI - Calcium phosphate bone cements for clinical applications. Part II: precipitate formation during setting reactions. AB - Calcium phosphate bone cements (CPBC) have been of great interest in medicine and dentistry due to their excellent biocompatibility and bone-repair properties. In this article, a review is presented of the scientific literature concerning precipitate formation during setting reactions of CPBCs. Firstly, the available information has been classified according to the intended final product or calcium phosphate formed during setting reactions. Taking the final product into account, a second classification has been made according to the calcium phosphates present in the original powder mixture. This is the most natural classification procedure because it is based on thermodynamic reasons supported by solubility diagrams for the calcium phosphate salts. By understanding the thermodynamics of calcium phosphate salts in an aqueous solution at room or body temperature it is possible to optimize the manufacturing technology involved in the production of CPBCs. Knowledge of the limitations of this thermodynamic approach opens up new possibilities in the search for CPBCs with better in vitro and in vivo properties for clinical applications. PMID- 15348167 TI - Electron microscopic evaluation of the effects of stress-shielding on maturation of the mid-substance and ligament-bone junction of the reconstructed anterior cruciate ligament in rabbits. AB - To analyze the effects of stress-shielding on graft maturation after the anterior cruciate ligament (ACL) reconstruction, autogenous ACL reconstruction using Achilles tendon was performed in rabbits. Two-end fixation with a ligament augmentation device (LAD), as a stress-shielding model (SS group), and pull-out fixation with Leeds-Keio artificial ligaments (L-K ligament), as a non-stress shielding model (non-SS group), were investigated. Intact ACL was used as the control. Specimens were harvested 6 mon postoperatively, and the analysis was focused on collagen fibril maturation in the mid-substance and bone-anchoring pattern in the bone tunnel, under light and transmission electron microscopy. The density of collagen fibrils in the control was lower than that in both experimental groups (p<0.01). The per cent collagen area in the control was higher than that in the SS (p<0.01) and the non-SS group (p< 0.05). The per cent collagen area in the non-SS group was higher than that in the SS group (p<0.05). At the ligament-bone interface, intracellular calcification of the degenerated chondrocytes occurred in the non-SS group, while in the SS group hydroxyapatite deposits were observed only in the extracellular matrix. These results show unfavorable influence of stress-shielding on graft maturation not only in the mid substance but also at the ligament-bone junction. PMID- 15348168 TI - Expression of caspase-8 and caspase-3 proteins in interface membranes from aseptically loose total hip arthroplasty. AB - The terminal events leading to periprosthetic osteolysis are multifactorial and modulation of this process after the stage of mediator release has been demonstrated futile. We demonstrated that ceramic induce macrophage apoptosis in vitro. More recently, we and others demonstrated the presence of apoptosis in interface membranes (IMs) from aseptically loose total hip arthroplasty (THA). The purpose of this study was to characterize the mechanisms leading to apoptosis in these pseudomembranes. Western blot analysis was used to characterize the expression of caspase-3, caspase-8, poly(ADP-ribose)polymerase (PARP), and p53 proteins in IM from 35 patients (40 specimens) with a mean age of 58 years (range, 28-88 years) at the time of revision. Tissue harvested at the time of routine hardware removal served as control. Our results show that caspase-3 and caspase-8 pro-enzymes were expressed in both control and IM tissues. Our results also showed that caspase-3 active fragment (17 kDa) as well as caspase-8 active fragment (18 kDa) were expressed in IMs but absent in control tissues. We also demonstrated that both the native PARP (113 kDa) and its proteolytic fragment (89 kDa) were present in osteolytic IMs. Control membranes expressed only the 113 kDa native form of PARP. The over-expression of caspase-3 caspase-8 active fragments and the presence of PARP fragment were observed on both the acetabular and femoral sides of the prostheses. Finally, our results showed the absence of p53 expression in both osteolytic IMs and control tissues. In conclusion, our results suggest that the caspase/PARP pathway plays an active role in the activity of IMs from aseptically loose THAs. PMID- 15348169 TI - pH-metric study of the setting reaction of monocalcium phosphate monohydrate/calcium oxide-based cements. AB - Hydraulic calcium phosphate cements (CPCs) that are used as osseous substitutes, set by an acid-base reaction between an acid calcium phosphate and a basic calcium salt (often a phosphate). In order to gain a better understanding of the setting of the monocalcium phosphate monohydrate-calcium oxide cement that we developed and in the aim to improve its mechanical properties, the setting reaction was studied by pH-metry. The two methods described in the literature were used. In the first, cement samples were prepared then crushed after different storage periods at 37 degrees C, 100% RH. The powder was then immersed in pure water with stirring and the pH was measured after equilibration. In the second technique, the starting materials were poured into water while stirring and the pH were followed over time. The two methods gave different results. The first procedure provided information concerning the pH of the surrounding liquid following the partial dissolution of the cement components, rather than any information about pH changes during setting. The second method is more appropriate to follow the pH variations during setting. In this second procedure, the effects of different parameters such as crushing time, stirring rate, liquid to-powder (L/P) ratio and temperature were investigated. These parameters may impact substantially on the shape and position of the pH=f(t) curves. One or three pH jumps were observed during the setting depending on the composition of the liquid phase. The time at which these pH jumps occurred depended on the pH of the liquid phase, the concentration of the buffer, the crushing of starting materials, the L/P ratio and the temperature. Good linear correlations were obtained (i) between the time of the pH jumps and the L/P ratio and the temperature and (ii) between the time of the first pH jump and the compressive strength and the final setting time of the cements prepared with different liquid phases. It may be assumed in view of these correlations that the results obtained in dilute solution may be extrapolated to the conditions of cement sample preparation and that the mechanical properties of the cement are directly related to the phenomena that occur at the first pH jump which corresponds to precipitation of dicalcium phosphate dihydrate. PMID- 15348170 TI - Bioactivity and mechanical properties of polydimethylsiloxane (PDMS)-CaO-SiO2 hybrids with different calcium contents. AB - Polydimethylsiloxane (PDMS)-CaO-SiO(2) hybrids with starting compositions containing PDMS/(Si(OC(2)H(5))(4)+PDMS) weight ratio=0.30, H(2)O/Si(OC(2)H(5))(4) molar ratio=2, and Ca(NO(3))(2)/Si(OC(2)H(5))(4) molar ratios=0-0.2, were prepared by the sol-gel method. The apatite-forming ability of the hybrids increased with increasing calcium content in the Ca(NO(3))(2)/Si(OC(2)H(5))(4) molar ratio range 0-0.1. The hybrids with a Ca(NO(3))(2)/Si(OC(2)H(5))(4) molar ratio range 0.1-0.2 formed apatite on their surfaces in a simulated body fluid (SBF) within 12 h. The hybrid with a Ca(NO(3))(2)/Si(OC(2)H(5))(4) molar ratio of 0.10 showed an excellent apatite-forming ability in SBF with a low release of silicon into SBF. It also showed mechanical properties analogous to those of human cancellous bones. This hybrid is expected to be useful as a new type of bioactive material. PMID- 15348171 TI - Creep behavior comparison of CMW1 and palacos R-40 clinical bone cements. AB - The restrained dynamic creep behaviors of two clinical bone cements, Palacos R-40 and CMW1 have been investigated at room temperature and body temperature. It was found that the two cements demonstrated significantly different creep deformations, with Palacos R-40 bone cement demonstrating higher creep strain than CMW1 bone cement at each loading cycle. For both cements, two stages of creep were identified with a higher creep rate during early cycling followed by a steady-state creep rate. The test temperature had a strong effect on the creep performance of the bone cements with higher creep rate observed at body temperature. The relationship between creep deformation and loading cycles can be expressed by single logarithmic model. The SEM examinations revealed that CMW1 bone cement is more sensitive to defects within the specimen especially to the defects at the edges of the specimen than Palacos R-40 bone cement. However, in the absence of micro-cracks or defects within the inner surface layer, the dynamic loading (at less than 10.6 MPa) is unlikely to produce micro-cracks in the CMW1 bone cement. The different behaviors between the two bone cements may be attributed to differences in chemical compositions and molecular weight distributions. PMID- 15348172 TI - The use of bifunctional polyethyleneglycol derivatives for coupling of proteins to and cross-linking of collagen matrices. AB - The realization of three-dimensional (3D) degradable matrices which slowly release bio-active components represents a major challenge in the field of tissue engineering. In this paper we report on the usage of commercially available bifunctional agents for both the covalent coupling of proteins to and the cross linking of collagen matrices. Proteins - horse radish peroxidase (HRP) was used as a model protein - were cross-linked with either a homobifunctional (disuccinimidyldisuccinatepolyethylene-glycol) or a heterobifunctional (N hydroxysuccinimidylvinylsulfonepolyethyleneglycol) agent. In the case of the heterobifunctional cross-linking agent the collagen matrices were previously modified with succinimidylacetylthioacetate in order to introduce sulfhydryl groups. As compared with control experiments a 10-fold and 50-fold increase of immobilized proteins were achieved with the homobifunctional and heterobifunctional cross-linker resp. The HRP-PEG conjugates demonstrated a better long-term stability as compared to the non-treated HRP. The effects of the cross-linking agents and the thiolation reagent succinimidylacetylthio acetate on the in vitro degradation of the collagen matrices by collagenase were also investigated. In particular the reaction with succinimidylacetylthio acetate appears to offer interesting opportunities both for coupling active proteins and modulating the degradation times of collagen matrices. PMID- 15348173 TI - Does sodium fluoride in bone cement affect implant fixation? Part I: bone tissue response, implant fixation and histology in nine rabbits. AB - The addition of sodium fluoride to poly (methyl-methacrylate) (PMMA) bone cement may theoretically improve the fixation of joint replacement. This hypothesis was tested in an animal model using nine mature healthy lop-eared rabbits. A femoral prosthesis was inserted in both knees to resurface the patellofemoral articulation. The same acrylic cement, with and without sodium fluoride, was randomised between the two sides for prosthetic fixation. Two screw shaped implants machined from cured rods of either cement were also inserted bilaterally into the proximal tibia. Qualitative and quantitative histomorphometry of the bone tissue response surrounding the cement in the femur and the intact tibial implants revealed similar results regardless of sodium fluoride addition. Six weeks after surgery removal, torque did not significantly differ between the two sides. Our findings indicate that addition of sodium fluoride to PMMA has little effect on implant stability and bone remodeling in rabbits in the short-term. PMID- 15348174 TI - Does sodium fluoride in bone cement affect implant fixation. Part II: evaluation of the effect of sodium fluoride additions to acrylic bone cement and the fixation of titanium implants in ovariectomized rabbits. AB - Bone integration of threaded implants made of cured polymethylmethacrylate containing sodium fluoride or commercially pure (c.p.) titanium were studied in normal and estrogen deficient New Zealand white rabbits. Nine had been ovariectomized through laparoscopy and nine served as controls. Four weeks after the ovariectomy two threaded implants made of cured bone cement with or without sodium fluoride addition were inserted in each tibia. One threaded commercially pure titanium implant was inserted in each patello-femoral joint flush to the cartilage. Six weeks after implant insertion measurement of the peak removal torque necessary to loosen the implants and light microscopical histomorphometrical investigations of tissue integration were performed. In the ovariectomized rabbits addition of sodium fluoride to the cement resulted in increased area of bone in the threads (p=0.04), but no corresponding effect could be noted in the controls. The removal torque was lower in the ovariectomized rabbits compared to the non-ovariectomized when comparing implant with sodium fluoride addition (p=0.02). The bone tissue response and the removal torque of the titanium implants were not influenced by ovariectomy in these rabbits. PMID- 15348175 TI - Anti-inflammatory prodrugs as plasticizers for biodegradable implant materials based on poly(3-hydroxybutyrate). AB - Salicylic and acetylsalicylic acid esters were tested as plasticizers for biodegradable poly(3-hydroxybutyrate) (PHB). The aim is the combination of plasticizing and anti-inflammatory properties in the fabrication of implant materials. Solution-cast films made of mixtures of PHB and 30% ester showed plasticization accompanied by a decrease of elastic modulus and an increase in elongation at break in comparison with pure PHB films. However, the number of usable plasticizers from the group of the salicylic acid and acetylsalicylic acid esters is limited. Short-chain derivatives are volatile while long-chain compounds tend to crystallize. In both cases PHB films embrittle within short time. Moreover, some derivatives show a fast release in an aqueous environment. As alternative nonsteroidal anti-inflammatory prodrugs arylpropionic acid esters were tested as plasticizers. The addition of ketoprofen ethyl ester led to PHB films with decreased brittleness. In summary, various esters of anti-inflammatory drugs show plasticizing effects on solution-cast PHB films comparable with those of commonly used citric acid esters. PMID- 15348176 TI - Micrometer level structural and chemical evaluation of electrodeposited calcium phosphate coatings on TA6V substrate by STEM-EDXS. AB - Hydroxyapatite (HA) coatings on titanium alloy substrates Ti6Al4V have been prepared in our laboratory by electrodeposition and hydrothermal synthesis. In this paper, the morphology, crystal size, porosity and Ca/P atomic ratio are investigated using scanning electron microscopy (SEM), scanning transmission electron microscopy (STEM), Raman microspectroscopy and X-ray energy dispersive spectroscopy (EDXS). The results obtained show that after being hydrothermally treated and calcined at high temperature, the electrodeposited brushite coating is converted into a stoichiometric hydroxyapatite having a crystal size which changes considerably from the surface to the substrate alloy. In addition, variation of the surface coating porosity as a function of the electrolyte temperature has also been carried out. PMID- 15348177 TI - Enhanced wear performance of ultra high molecular weight polyethylene crosslinked by organosilane. AB - Ultra high molecular weight polyethylene (UHMWPE) crosslinked by organosilane was thermal compression molded. The organosilane used was the tri-ethyloxyl vinyl silane. Its gelation, melting behavior, crystallinity, mechanical and wear resisting properties were systematically investigated. The results showed that the gel ratio of UHMWPE increases with the incorporation of organosilane. At a low content of organosilane, the melting point and crystallinity of the crosslinked UHMWPE increase, and hence the mechanical and wear-resisting properties are improved. However, at a high content of organosilane, these performances of the crosslinked UHMWPE become worse. At 0.4 phr silane, the wear resistance of crosslinked UHMWPE reaches its optimum value. PMID- 15348178 TI - Osseointegration and osseoconductivity of hydroxyapatite of different microporosities. AB - The aim of this study was to determine the relationship between changes in microporosity and the osseointegration and the osseoconductivity of hydroxyapatite (HAp). HAp cylinders were manufactured by a combination of uniaxial powder pressing and different sintering conditions, with different percentages of microporosity: HAp-I with 3.96+/-0.75% microporosity, HAp-II with 15.66+/-1.60% microporosity, and HAp-III with 29.72+/-0.69% microporosity. These HAp cylinders were surgically implanted in rabbit femurs. After 8 and 12 weeks, the femurs were removed, fixed, sectioned, ground, and stained by Stevenel's blue/Van Gieson for light microscopy and histomorphometry. Some ground sections were routinely processed for SEM. The osseointegration and the osseoconductivity were determined by means of image analysis and the data were submitted to ANOVA. In all cases the cortical bone was repaired and the HAp facing the medullary canal was lined with endosteum, which in some areas exhibited thin bone tissue formation. SEM observations showed no differences in the morphology of tissue-HAp interfaces for the three different porosities of HAp. There were no statistical differences between the groups related to either osseointegration or osseoconductivity. These results suggest that neither osseointegration nor osseoconductivity of HAp are influenced by changes in HAp microporosity. PMID- 15348179 TI - Influence of pressurization on flexural strength distributions of PMMA-based bone cements. AB - Flexural strength distributions of standard viscosity and low viscosity bone cements based on Polymethylmethacrylate were obtained by testing the materials in four-point bending according to the ISO 5833 protocol. The cement dough was poured into a mold and was allowed to cure at atmospheric pressure. An additional set of specimens of the standard viscosity cement was prepared under pressure while the cement dough was polymerizing in the mold. Following preparation, test specimens were stored in a 37 degrees C water bath for 48 h. The two-parameter Weibull model, which was used to analyze the data, gave a good representation of the fracture loads distribution. Low viscosity cement displayed a higher mean flexural strength and a slightly lower data scatter than standard viscosity cement. The mean flexural strength of the cement increased about 60% when pressure was applied compared with the same material cured at atmospheric pressure. The Weibull modulus, m, characterizes the scattering in the measured values of strength. For the cement prepared at atmospheric pressure the m value was 8.6 while for the cement cured under pressure it was 12.3, which reveals a reduction in the data scatter. The cement tested in four-point bending displayed lower mean flexural strength compared with the cement tested in three-point bending. The influence of the load type upon the mean flexural strength was satisfactory predicted by Weibull model. PMID- 15348180 TI - PIXE micro-beam mapping of metals in human peri-implant tissues. AB - Previous investigations did not agree about the possible presence of titanium and other metals in the tissues around endosteal dental implants and joint prostheses. Indeed, while some authors reported diffusion of metals into the tissues, some others did not find evidence of this phenomenon. In the present study, four dental titanium implants, removed with the surrounding tissues from patients at various time intervals after the insertion, were studied by means of the micro-beam proton-induced X-ray emission (PIXE micro-beam) technique, which draws maps showing the tissue distribution of elements with a detection limit of about 1 ppm. One implant was built in commercially pure titanium, two others in titanium coated with titanium plasma spray, and the fourth in Ti-Al-V alloy. Their composition was confirmed by the PIXE micro-beam analyses. The removed samples were embedded in epoxy and processed with a cutting-grinding appliance, mounted on plastic holders, and ground up to a thickness of about 35 microm. Optical microscope examinations were also carried out, to compare the optical findings with the elemental maps obtained with the PIXE micro-beam. One implant, removed after 70 days because the patient had developed peri-implantitis, had some inflammatory soft tissue attached, with no evidence of metal leakage. The other three implants had been removed after 6, 7 and 9 years of valid clinical service, because of the fracture of the prosthetic abutment or the implant stem. At the optical microscope, all these fixtures were embedded in mature bone. The elemental maps indicated small titanium deposits in about 5% of the bone bordering the implants, while aluminum, when present in the fixture, leaked diffusely into the surrounding bone and vanadium was not found in the tissues. These results suggest that titanium may be found occasionally in peri-implantar tissues, but has very little tendency to spread, while the presence of aluminum in the implant alloy may cause an important leakage of this metal. PMID- 15348181 TI - Effects of in vitro degradation on properties of poly(DL-lactide-co-glycolide) pertinent to its biological performance. AB - Poly(DL-lactide-co-glycolide) (PLGA) is a widely used biodegradable polymer. In this study, effects of in vitro degradation on its dynamic mechanical properties, thermal behavior, molecular weight, fluid uptake and dimension stability were investigated. The in vitro experiment was conducted at 37 degrees C in phosphate buffer solution with pH=7.4 in a dynamic incubator. The dynamic mechanical experiments were performed on rectangular specimens under three point bending. For an aging period of 6 weeks, the dynamic mechanical properties were found to decrease significantly with the increased time. The absorbed fluid had a significant effect on polymer storage modulus. The molecular weight decreased with aging time, and was found to be different at the center and the surface, being higher at the center in the beginning and lower thereafter. The polydispersity did not change significantly with degradation. The fluid uptake and specimen geometric dimension increased proportionally with aging time. The glass transition and thermal decomposition temperatures decreased with the increased degradation time. PMID- 15348182 TI - Self-reinforcement and hydrolytic degradation of amorphous lactic acid based poly(ester-amide), and of its composite with sol-gel derived fibers. AB - The self-reinforcing and hydrolytic degradation of an amorphous poly(ester-amide) (PEA) based on lactic acid have been studied and compared with those of poly-L lactide (PLLA). The studied PEA-rods were self-reinforced (SR) by solid-state die drawing resulting double shear strength. The hydrolytic degradation of PEA was studied during exposure to phosphate buffered saline at pH 7.4 and at 37 degrees C for 18 weeks. The degradation and mechanical properties of PEA were also followed in a self-reinforced composite structure consisting of PEA and sol-gel derived SiO(2)-fibers (SGF, 8 wt %). The hydrolytic degradation of the SR-PEA rods with and without SG-fibers was significantly faster than that of SR-PLLA rods. The weight average molecular weight (Mw) of PEA decreased by 90% from the initial Mw during the first 6 weeks in hydrolysis, when the Mw of the PLLA decreased by 10%. PMID- 15348183 TI - Electrochemical polishing of 316L stainless steel slotted tube coronary stents. AB - Surface smoothness is one of the properties determining the performance of stents. Therefore, surface polishing shows its importance in the exploitation and production of stents. The present study explores electrochemical polishing of 316L stainless steel slotted tube coronary stents produced by laser cutting. Acid pickling was also studied as a pre-treatment of electrochemical polishing of the stents to remove the slag (oxides) formed in the production of the stents. Meanwhile, removal of the material was measured as well, caused by both acid pickling and electrochemical polishing processes. It is found that the slag formed on the surface of stents due to the laser cutting production process could be removed by means of acid pickling. Electrochemical polishing results in a smooth stent surface. Meanwhile, both acid pickling and electrochemical polishing applied in the present study have a proper removal of the stent material. PMID- 15348184 TI - Studies on the effect of electron beam radiation on the molecular structure of ultra-high molecular weight polyethylene under the influence of alpha-tocopherol with respect to its application in medical implants. AB - Ultra-high molecular weight polyethylene (UHMW-PE) is being used successfully for articulating surfaces in joint endoprostheses, especially for cups of total hip endoprostheses. Sintered specimens containing various amounts of alpha-tocopherol (vitamin E) as a biocompatible stabilizer, were irradiated in nitrogen atmosphere as well as in air with various dosages of electron beam radiation. Size exclusion chromatography (SEC) was used to analyze the soluble fractions of the UHMW-PE samples according to their molecular weight distribution prior to and after irradiation. In nitrogen atmosphere the radiation-induced crosslinking showed to be dependent on the added amount of alpha-tocopherol in the sintered specimens. With an increasing content of alpha-tocopherol, the stabilizer acted as a scavenger for free radicals. Thus, the crosslinking was more and more hindered. The same effect was observed on the samples irradiated in air, where, in addition to the crosslinking process, oxidative molecular degradation occurred. The highest extent of crosslinked material was yielded with unstabilized samples in nitrogen atmosphere. PMID- 15348185 TI - The interactions of bisphosphonates in solution and as coatings on hydroxyapatite with osteoblasts. AB - Aseptic loosening is one of the major causes of failure of artificial hip joints, and it can occur for several reasons, including osteolysis of the bone tissue in response to stress shielding or cellular reactions to wear debris. Any treatment of the prosthesis which could minimize the osteolytic response of bone tissue may be able to extend the life-time of the implant. Bisphosphonates are potent inhibitors of osteoclastic bone resorption, and they bind avidly to hydroxyapatite (HA). Coating the prostheses with bisphosphonates may therefore inhibit osteolysis. We have investigated the potential for this approach by determining whether bisphosphonates interact with osteoblasts in vitro. The effects of pamidronate (P), clodronate (C), and etidronate (E) in solution and when coated onto HA were investigated. P inhibited protein and collagen syntheses potently when in solution, but not after being bound to HA. When bound to HA, both P and C increased DNA, protein and collagen syntheses of osteoblasts and may encourage the osseointegration of implants. The pharmacological effects of the bisphosphonates studied altered dramatically after binding to HA. This must be fully investigated before this approach to prolonging prostheses stability can be evaluated. PMID- 15348186 TI - Boundary conditions during biaxial testing of planar connective tissues. Part 1: dynamic behavior. AB - Current mechanical testing methods used to determine the biaxial properties of planar connective tissues may lead to artifactual observations of material behavior. The method of sample gripping affects the constraint on the extracellular fibers at the bounds of the sample. This applied constraint not only affects how the load is transferred to the sample, but also how the load is transmitted throughout the rest of the material - thereby influencing the resulting mechanical behavior of the tissue. In this study, we compared the dynamic biaxial mechanical response of pericardial tissue samples under two different gripping methods: (i) the common method of suturing sample edges and (ii) a new biaxial clamping method. Tissue samples were repeatedly testing using both gripping methods under the same conditions. The tissue samples appeared to be stiffer and less extensible when mechanically tested with clamped sample edges, as opposed to when tested with sutured sample edges. Thus, the influence of the sample boundaries affected the response of the material - precisely the situation to be avoided for reliable material testing. This casts doubt on whether any in vitro mechanical testing method can used to determine the "real" properties of the tissue since the boundary conditions of the tissue in situ are presently unknown. PMID- 15348187 TI - Novel starch thermoplastic/Bioglass composites: mechanical properties, degradation behavior and in-vitro bioactivity. AB - The present research aims to evaluate the possibility of creating new degradable, stiff and highly bioactive composites based on a biodegradable thermoplastic starch-based polymeric blend and a Bioglass filler. Such combination should allow for the development of bioactive and degradable composites with a great potential for a range of temporary applications. A blend of starch with ethylene-vinyl alcohol copolymer (SEVA-C) was reinforced with a 45S5 Bioglass powder presenting a granulometric distribution between 38 and 53 microm. Composites with 10 and 40 wt % of 45S5 Bioglass were compounded by twin-screw extrusion (TSE) and subsequently injection molded under optimized conditions. The mechanical properties of the composites were evaluated by tensile testing, and their bioactivity assessed by immersion in a simulated body fluid (SBF) for different periods of time. The biodegradability of these composites was also monitored after several immersion periods in an isotonic saline solution. The tensile tests results obtained indicated that SEVA-C/Bioglass composites present a slightly higher stiffness and strength (a modulus of 3.8 GPa and UTS of 38.6 MPa) than previously developed SEVA-C/Hydroxylapatite (HA) composites. The bioactivity of SEVA-C composites becomes relevant for 45S5 amounts of only 10 wt %. This was observed by scanning electron microscopy (SEM) and confirmed for immersion periods up to 30 days by both thin-film X-ray diffraction (TF-XRD) (where HA typical peaks are clearly observed) and induced coupled plasma emission (ICP) spectroscopy used to follow the elemental composition of the SBF as function of time. Additionally, it was observed that the composites are biodegradable being the results correlated with the correspondent materials composition. PMID- 15348188 TI - Effect of heat treatment with the mould on the super-elastic property of Ti-Ni alloy castings for dental application. AB - Tensile property of Ti-50.85Ni (mol %) alloy castings was investigated quantitatively in relation to the thermal behavior accompanied with phase transformation to evaluate the effect of heat treatment after casting with the mould in air. The heat treatment temperature was 713 or 773 K, and the period was 0.9, 1.8, or 3.6 ks. Apparent proof stress of the castings decreased with increasing period of heat treatment, and the decrease was larger with the treatment at 773 K. Residual strain also decreased by the heat treatment, however, it was low with the treatment for relatively short period, i.e. 713 K 0.9 and 1.8 ks, and 773 K-0.9 ks treatments. From the thermal behavior measured by differential scanning calorimetry (DSC), the ascent in the transformation temperatures and the increase in the thermal peak height appeared to influence the changes in the tensile property. These changes by heat treatment were believed to be effective to utilize more flexibility, less stress and less permanent deformation in dental castings. PMID- 15348189 TI - Computed tomography assessment of alveolar filling with an injectable bone substitute. PMID- 15348190 TI - Electrochemical behavior of nitinol alloy in Ringer's solution. AB - Corrosion behavior of Nitinol (nickel titanium) alloy has been examined in physiological Ringer's solution by means of electrochemical techniques (open circuit potential measurement, potentiodynamic and potentiostatic polarizations) and capacitance measurements. The results show that the passive film is stable at open-circuit condition, but many of anodic current transients are observed at potentiodynamic and potentiostatic polarizations. The thickness of passive film decreased because of the occurrence of dissolution. PMID- 15348191 TI - In-vitro forming of calcium phosphate layer on sol-gel hydroxyapatite-coated metal substrates. AB - In-vitro deposition of calcium phosphate layer (CPL) on metallic substrate requires special surface preparation in order to provide an interfacial bond. In this work 316 stainless steel surface is modified through deposition of a thin film ( approximately 0.5 microm) of sol-gel hydroxyapatite (SG-HA). This well bonded film acts as an intermediary and nucleation surface of the CPL film. The SG-HA films were annealed at 375 degrees C (samples coded 375-ACS) and 400 degrees C (400-ACS) to achieve different crystallinity of the films, and thus to affect and study the CPL nucleation process. The CPL growth was investigated in terms of deposition kinetics and microstructural development. A deposition rate of dense CPL of about 0.43 microm/day was achieved on the crystallized film of 400-ACS, and 0.22 microm/day of porous CPL on amorphous 375-ACS. A compositional variation of Ca/P ratio across the CPL film thickness (400-ACS) was observed. Lower Ca/P ratio of 1.2 was detected near the substrate-CPL interface and about 1.5 near the solution-CPL interface. Infrared analysis showed the CPL to be of apatitic calcium-deficient structure. Kinetic model explaining the advancement of the CPL upon the in-vitro immersion is proposed. PMID- 15348192 TI - The in vitro antibiotic release from anti-washout apatite cement using chitosan. AB - The in vitro antibiotic release from anti-washout apatite cement using chitosan (aw-AC(chi)) was investigated in a preliminary evaluation. Flomoxef sodium was employed as the antibiotic and was incorporated into the powder phase aw-AC(chi) at up to 10%. The setting times were measured for aw-AC(chi) containing various amounts of flomoxef sodium. X-ray diffraction (XRD) analysis was also conducted for the identification of products. To evaluate the drug release profile, set aw AC was immersed in saline and the released flomoxef sodium was determined at regular intervals. The setting time was prolonged slightly with the addition of flomoxef sodium. The difference at 10% flomoxef sodium (0% vs. 10%) was not significant (p>0.05), and can be negligible in clinic. The XRD analysis revealed that formation of hydroxyapatite (HAP) from aw-AC(chi) was reduced, even after 24 h, when the aw-AC(chi) contained flomoxef sodium at 8% or more. The flomoxef sodium release from aw-AC(chi) showed the typical profile observed in skeleton type drug delivery system (DDS). Changing the concentration of chitosan can control the rate of drug release from aw-AC. Therefore, we conclude that aw AC(chi) is a good candidate for potential use as a DDS carrier that may be useful in surgical operations. PMID- 15348193 TI - Age-hardening behaviors and grain boundary discontinuous precipitation in a Pd free gold alloy for porcelain bonding. AB - Isothermal age-hardening behaviors at 400 degrees and 450 degrees C and discontinuous precipitation reaction at 450 degrees C in a commercial Pd-free gold alloy for porcelain bonding were investigated by hardness testing, X-ray powder diffraction, and light microscopy. Variations of electrical resistivity during continuous heating and cooling processes were also measured. The alloy exhibited pronounced age-hardening in the early stage of aging and the maximum hardness exceeded twice that of the solution-treated sample. Precise lattice parameter measurements and investigations of full width at half maximum values for the X-ray Bragg reflections implied that nonuniform strains due to the pre precipitation or zone formation was responsible for the quick and pronounced age hardening at 450 degrees C. Discontinuous precipitation reaction, producing a mixture of a small amount of Pt(3)In-phase with the L1(2)-type superstructure and a large amount of (Pt, In)-depleted solid solution, started at grain boundaries in the late stage of aging process at 450 degrees C. The growth of the grain boundary discontinuous precipitates toward the intragrain area led to a gradual decrease in hardness of the alloy. PMID- 15348194 TI - Fabrication and characterization of hydroxyapatite reinforced with 20 vol % Ti particles for use as hard tissue replacement. AB - Hydroxyapatite(HA)-based composite reinforced with 20 vol % titanium (Ti) particles was fabricated by hot pressing based on the studies of the structural stability of HA phase in HA-Ti composite by means of FTIR spectrometry and X-ray diffractometry. The mechanical properties and biological behaviors of the composite were investigated by mechanical and in vivo studies. The existence of Ti metal phase can promote the dehydration and decomposition of HA ceramic phase into the more stable calcium phosphate phases, such as alpha-Ca(3)(PO(4))(2) (alpha-TCP) and Ca(4)O(PO(4))(2) at high temperatures. Comparing with pure HA ceramic manufactured under the same conditions, HA-20 vol % Ti composite with higher fracture toughness (0.987 MPa m(1/2)), bending strength (78.59 MPa), work of fracture (12.8J/m(2)), porosity (9.8%) and lower elastic modulus (75.91 GPa) is more suitable for use as hard tissue replacement. Crack deflection is the chief toughening mechanism in the composite. Histological evaluation by light microscope shows HA-20 vol % Ti composite implant could be partially integrated with newborn bone tissues after 3 weeks and fully osteointegrated at 12 weeks in vivo. The excellent biological properties of HA-20 vol % Ti composite may be contributed to the coexistence of high porosity and the decomposition products of HA phase in the composite. PMID- 15348195 TI - The cross-linkage effect of hydroxyapatite/collagen nanocomposites on a self organization phenomenon. AB - Hydroxyapatite(HAp)/collagen nanocomposites were prepared by a coprecipitation method controlling the degree of cross-linkage between collagen molecules using glutaraldehyde. The precipitates filtered were dried in a freeze drier or naturally dried in the air at 25 degrees C. The naturally dried cakes had open channels of 5-15 microm in diameters, which were three-dimensionally and regularly developed over the whole samples, and showed a pretty good mechanical strength. The channels that were formed at spaces among the HAp/collagen particles, cross-linked one another, which had been filled up with water before its evaporation. The ordering state of the open channels depended on the degree of cross-linkage with glutaraldehyde; the optimal self-organized state was found when 30 molecules of glutaraldehyde were added per collagen molecule, though an excess amount of glutaraldehyde suppressed the appearance of the ordered state. From SEM and FT-IR measurements, it was indicated that the self-organization in the HAp/collagen nanocompsites continuously occurred during the drying process together with the removal of water and the increase of the density. PMID- 15348196 TI - Material properties of various cements for use with vertebroplasty. AB - The purpose of the current study was to measure the material properties of various cements prepared per manufacturers' recommendations and of cements modified according to compositions developed by clinicians with experience performing vertebroplasty. Cement was prepared, cast to form cylindrical specimens, and tested in compression. The optical density of specimens from the various cement preparations was measured. Batches of Simplex P and Cranioplastic cement were also prepared with increased concentrations of BaSO4 (20% and 30%; and 10%, 20% and 30%, respectively) to evaluate the effect of additional BaSO4. Compressive modulus values for polymethylmethacrylate cements ranged from 2-2.7 GPa; some differences were significant (p<0.05). Compared with polymethylmethacrylate cements, Orthocomp exhibited almost twice the compressive modulus and 2-3 times the strength values. Increasing the BaSO4 concentration in Simplex P and Cranioplastic significantly (p<0.05) affected their material properties; however, it is unknown if these changes in material properties are clinically important. Optical density increased as a function of concentration of the opacifying agent added. The current study provides clinicians with information on changes in the material properties of bone cements when the compositions are altered in a manner consistent with the practice of vertebroplasty. PMID- 15348197 TI - An investigation into the architecture and composition of a urinary calculus. AB - A bladder stone of diameter of 16 mm was sectioned to reveal an internal structure showing some complexity. Visually apparent were concentric rings divided into two principal zones. The outer zone comprised large single interlocking crystals, but the inner zones consisted of a much more fragile environment in which large single crystals were embedded in a much finer matrix. More detailed analysis was performed on the stone's layers using X-ray diffraction which is capable of identifying the crystallites but may also be used to elucidate some of the crystallite characteristics and thereby provide evidence for the growth environment. Proton induced X-ray emission was carried out at a number of points along a radius of the sample to examine the distribution of key trace elements. A possible mechanism for the influence of this banding on the fragility of the stone is discussed. PMID- 15348198 TI - Synthesis and characterization of a novel double crosslinked hyaluronan hydrogel. AB - Hyaluronan has great potential in medicine as a biomaterial. However, in its native form, hyaluronan is rapidly metabolized in vivo by free radicals and enzymes such as hyaluronidase, and it is highly soluble. Various methods have been adopted therefore, to modify the physicochemical properties of hyaluronan, while maintaining biocompatibility, and thereby widen its spectrum of therapeutic applications. Hyaluronan has four reactive groups (acetamido, carboxyl, hydroxyl and the reducing end) available for crosslinking to itself or other polymers. Using a variety of crosslinking agents, researchers have developed a host of crosslinked hyaluronan derivatives with an increased in vivo residence time. This chemical modification has enabled the production of gels and films, which can be used in applications such as the prevention of post-surgical adhesions, wound healing and dermal augmentation. We have found that if hyaluronan is crosslinked to itself, or to other polymers (either synthetic or biopolymer), in two stages, then a high degree of crosslinking is achieved, conferring improved biostability. In each of the two stages, the same crosslinking agent is used, but different functional groups are bound by altering the reaction conditions. The novel process can be tailored to yield water insoluble gels and films with a broad range of physical and chemical characteristics, and greater resistance to degradation by hyaluronidase and free radicals. These derivatives are currently undergoing biocompatibility testing, and should ultimately lead to a series of innovative second-generation medical products. PMID- 15348199 TI - Curing characteristics of acrylic bone cement. AB - Commercial acrylic bone cements are supplied as two components, a polymer powder and a liquid monomer. Mixing of the two components is followed by a progressive polymerization of the liquid monomer to yield a solid mass, a high level of heat being generated during this exothermic reaction. The exposure of bone to high temperatures has led to incidences of bone necrosis and tissue damage, ultimately resulting in failure of the prosthetic fixation. The aim of this study was to determine the thermal properties of two acrylic bone cements as they progress through their polymerization cycles. It was also felt that there was a need to quantify the variations in the curing characteristics as a function of preparing bone cement by different techniques, hand mixing and vacuum mixing. A number of parameters were calculated using the data gathered from the investigation: peak temperature, cure temperature, cure time, and the cumulative thermal necrosis damage index. The results show the temperature profile recorded during polymerization was lowest when the cement was prepared using the Howmedica Mix Kit I system: 36 degrees C for Palacos R and 41 degrees C for CMW3 respectively. When the acrylic cements were prepared in any vacuum mixing system there was evidence of an increase in the cure temperature. The main factor that contributed to this rise in temperature was an imbalance in the polymer powder : liquid monomer ratio, there was a high incidence of unmixed powder visible in the mixing barrel of some contemporary vacuum mixing devices. Observing the thermal characteristics of the polymethyl methacrylate (PMMA) bone cements assessed, it was found that particular formulations of bone cements are suited to certain mixing methodologies. It is vital that a full investigation is conducted on a cement mixing/delivery system prior to its introduction into the orthopaedic market. PMID- 15348200 TI - Processing and characterization of porous alumina scaffolds. AB - Bioceramic materials are used for the reconstruction or replacement of the damaged parts of the human body. In this study an improved procedure is described for producing ceramic scaffolds with controlled porosity. Bioinert alumina ceramic was used to make porous scaffolds by using indirect fused deposition modeling (FDM), a commercially available rapid prototyping (RP) technique. Porous alumina samples were coated with hydroxyapatite (HAp) to increase the biocompatibility of the scaffolds. Initial biological responses of the porous alumina scaffolds were assessed in vitro using rat pituitary tumor cells (PR1). Both porous alumina and HAp coated alumina ceramics provided favorable sites for cell attachments in a physiological solution at 37 degrees C, which suggests that these materials would promote good bonding while used as bone implants in vivo. Based on these preliminary studies, similar tests were performed with human osteosarcoma cells. Cell proliferation studies show that both the ceramic materials can potentially provide a non-toxic surface for bone bonding when implanted in vivo. PMID- 15348201 TI - Poly(vinyl alcohol) hydrogels as hydrophilic matrices for the release of lipophilic drugs loaded in PLGA nanoparticles. AB - Poly(vinyl alcohol) (PVA) hydrogels prepared by a freeze-thawing procedure were evaluated as matrices for the release of water-insoluble drugs such as dexamethasone. As it is impossible to directly entrap a lipophilic drug into a hydrophilic matrix, a novel mechanism has been designed based on producing biodegradable nanoparticles loaded with the drug, that could then be entrapped into the hydrogels. Nanoparticles were prepared by a solvent evaporation technique using a biodegradable copolymer of poly(lactic acid)-poly(glycolic acid) (PLGA). The effects of several processing parameters on particle properties were investigated. The drug release from free nanoparticles was compared to that from the nanoparticles entrapped into the PVA matrices. It was observed that the release profile of the drug is not significantly affected by the PVA matrix. A correlation was found between the amount of drug released and the PVA concentration in the hydrogels: the percentage of drug released, as a function of time, decreased by increasing PVA concentration, indicating that PVA concentration can be used as a tool in modulating the release of the drug. PMID- 15348202 TI - Observation of fluorapatite formation under hydrolysis of tetracalcium phosphate in the presence of KF by means of soft X-ray emission and absorption spectroscopy. AB - The effect of fluoride on the hydrolysis of tetracalcium phosphate (TTCP; Ca4(PO4)2O) in 0.1 mol/l KH2PO4 containing 62-83 mmol/l KF was studied with the help of X-ray fluorescence measurements. Fluorine X-ray emission and absorption spectra of the final product of hydrolysis and reference samples (CaF2 and Ca5(PO4)3F) were measured at Beamline BL-2C of Photon Factory (PF, Tsukuba). Based on these measurements we concluded that hydrolysis of TTCP in the presence of KF converts it into fluorapatite. Formation of CaF2, which is often found in the hydrolysis of hydroxyapatite at high fluoride concentration, was not observed. PMID- 15348203 TI - Surface characterization and platelet adhesion studies of aliphatic polyurethanes grafted by fluorocarbon oligomers: effect of fluorocarbon chain length and carboxylic acid group. AB - The surfaces of aliphatic polyurethane films, which were synthesized by 1,6 hexamethylene diisocyanate, poly(tetramethylene glycol) and 1,4 butanediol, were modified by grafting different chain length of fluorocarbon oligomers. The fluorocarbon oligomers on polyurethane surfaces were terminated with trifluorocarbon or carboxylic acid functionality. The alkyl groups were also grafted onto polyurethane surfaces for comparison. The surface characterization and platelet-contacting property were studied using electron spectroscopy for chemical analysis (ESCA), static contact angle analysis and in vitro platelet adhesion experiments. The effects of fluorocarbon oligomers and their terminal functionalities are discussed. The ESCA results demonstrate the fluorocarbon enrichment at the outmost layer in fluorocarbon oligomer grafted polyurethanes. The fluorocarbon content at the surface increases with increasing the chain length of fluorocarbon oligomers. The fluorocarbon oligomer grafted polyurethanes exhibit highly hydrophobic surfaces, while alkyl groups grafted polyurethanes show relatively hydrophobic surfaces compared with the untreated polyurethane. The in vitro platelet adhesion experiments indicated that the fluorocarbon oligomer and carboxylic acid functionality significantly reduced the number and the degree of activation of the adherent platelets. PMID- 15348204 TI - The cytotoxicity of chromium in osteoblasts: effects on macromolecular synthesis. AB - Exposure of an immortalized rat osteoblast cell line, FFC cells, to Cr VI resulted in inhibition of protein, DNA and RNA synthesis. Protein synthesis (3H leucine incorporation) was most sensitive. There was no inhibition of the incorporation of 3H-proline into collagen at the concentrations which inhibited general protein synthesis (1 microM), but synthesis of extracellular collagen fibers was markedly decreased by concentrations of 0.5 microM Cr VI and above. This indicates that some aspect of the post-translational processing of the collagen fibers is sensitive to Cr VI inhibition. Collagen fiber formation was not inhibited by Cr III (which does not penetrate the cell membrane) or when Cr VI was reduced to Cr III extracellularly. This suggests that the Cr VI inhibits an intracellular stage of post-translational collagen processing. Both Cr VI and Cr III inhibit collagenase activity, the former being more potent but less efficacious. Our results suggest that leakage of chromium ions from orthopedic implants may cause a decrease in the proliferation and infiltration of osteoblasts around the implant, and a reduction in the synthesis and altered turnover of collagen in extracellular matrix. These effects will influence the osseointegration of implants, the osteolytic response, and ultimately the stable life-time of the implants. PMID- 15348205 TI - Controlled drug release for oral condition by a novel device based on ethylene vinyl acetate (EVA) copolymer. AB - The application of drug delivery systems in oral environment is relatively a new area of research with the exception of release of fluoride ions from polyalkenoate cements and their predecessor silicate cements. The present study addresses development of a novel device based on ethylene vinyl acetate copolymer (EVA), a biocompatible material which enables constant drug release over several days to treat oral infections. Drugs incorporated in EVA included tetracycline, minocycline and nystatin together with combinations (C) of nytatin-tetracycline (1 : 1) and nystatin-minocycline (1 : 1). Polymer casting solutions were prepared by dissolving EVA and the drugs in the ratio of 10 : 1 in 70 ml of dichloromethane at 38 degrees C for 6 h. Thin square films of 3x3 cm and 1 mm thickness were cut from the dry sheet obtained by solvent evaporation. Drug loaded samples were extracted for a minimum of 15 days in 10 ml medium (water or water/ethanol (1 : 2) or 0.9% saline solution) which is replaced daily. Spectral measurements were made to follow changes in optical densities (OD) during release kinetics. Analysis of the data revealed that among all the drugs tested tetracycline exhibited the highest release rate (56.15microg/cm2/day) and % cumulative release (27.92). The observed enhanced values may be interpreted as due to the channels formed due to changes in free volume (microvoids). In case of nystatin-minocycline combination, the observed increased values of release rates and percent cumulative release, may be attributed to the swelling component or channels or relative hydrophobic interactions. Initial "burst" effects due to liberation of surface-bound drug molecules were observed with reference to all the three drugs and the combinations of drugs studied. Among all the drugs, minocycline exhibited the least "burst" effect suggesting that the drug is more homogeneously distributed in the copolymer. Drug loaded EVA thermoplastic copolymer may provide a favorable therapeutic material for the development of a novel, local treatment for oral, mucosal and periodontal infections. PMID- 15348206 TI - Vibrational and thermal study on the in vitro and in vivo degradation of a bioabsorbable periodontal membrane: Vicryl Periodontal Mesh (Polyglactin 910). AB - Fourier transform Raman (FT-Raman), attenuated total reflection/Fourier transform infrared (ATR/FT-IR) spectra and differential scanning calorimetry (DSC) measurements were performed on a biodegradable periodontal membrane, the Vicryl periodontal mesh, in order to study its in vitro and in vivo degradation mechanism and kinetics. The hydrolitic in vitro degradation was investigated in two aqueous media: a saline phosphate buffer (SPB, pH=7.4) and a 0.01 M NaOH solution. Moreover, a membrane implanted in vivo for 4 weeks for treatment of contiguous vertical bony defects, was examined. Vibrational and thermal measurements show that the Vicryl membrane presents a semicrystalline structure. It degrades faster in the NaOH solution than in the SPB and degradation occurs heterogeneously with a progressive increase in the percentage of crystallinity and shortening of the polymeric chains both in vitro and in vivo. The trends of % weight loss and IR I627/I1415 intensity ratio (identified as a marker of crystallinity) are discussed in comparison with the DSC results. The IR I627/I1415 intensity ratio and Xc% allow to determine the % weight loss undergone by the membrane degraded in vivo. The result obtained shows that the Vicryl membrane degrades faster in vivo than in vitro with the formation of oligomers which are more easily absorbed by the surrounding tissues than they are soluble in the degradation media examined. PMID- 15348207 TI - Shear-load carrying capacities of the distal rat femora after osteotomy fixed with self-reinforced polyglycolic acid and poly-L-lactic acid pins. AB - Distal femora of 40 rats were osteotomized and fixed with self-reinforced polyglycolide (SR-PGA) and self-reinforced polylactide (SR-PLLA) pin 2.0 mm in diameter and 15 mm in length. The shear-load carrying capacities of the osteotomized bones were compared with each other and with the intact control rat distal femora of the same age of 20 pairs. The follow-up times were 1, 3, 6, 12, 24, 36, 48, and 52 weeks. After killing all operated and control femora were examined macroscopically and radiographically. The shear-load carrying capacities reached their highest values at 24 weeks in the SR-PGA-fixed specimens, after that decreasing to the level where they remained. In the SR-PLLA-fixed specimens the strength values of the pins increased after three weeks, but there was a decrease at 24 weeks. After that the shear-load carrying capacities started to raise because of the influence of the healed osteotomy. In the control bones the shear-load carrying capacities were weaker than in the SR-PGA-and SR-PLLA-fixed specimens except at three weeks, as the osteotomies had not yet healed. During the whole follow-up period the mean shear-load carrying capacity of the SR-PGA fixed specimens was 199.1 N, in the SR-PLLA-fixed specimens 214.6 N, the corresponding value of the control specimens being 148.2 N. PMID- 15348208 TI - The crystallization of calcium carbonate on sodium cholate. AB - Recent studies in the bibliography showed that calcium carbonate was the major constituent (77.8%) in gall stones, and the polymorph calcite was at 62.5% of the cases examined. The kinetics of crystallization of calcite on sodium cholate has been studied using the constant composition technique. Analysis of the initial rates as a function of the solution supersaturation, according to the classical nucleation theory, yielded a value of 33mJm(-2) for the surface energy of the growing phase and a five-ion cluster, forming the critical nucleus. The apparent order for the calcite crystallization was found to be 4.5+/-0.7 indicative of a surface nucleation mechanism. The formation of calcite may be initiated through the interaction of Ca2+ ions with the negative end of the C=0 bond of the sodium cholate molecule. PMID- 15348209 TI - Characterization of proteins and fibroblasts on thin inorganic films. AB - The ability of biomaterial surfaces to regulate cell behavior requires control over surface chemistry and material microstructure. One of the goals in the development of silicon-based biomedical devices such as biosensors or drug delivery systems is improved biocompatibility which may be achieved through the deposition or adsorption of thin films. In this study, films of single crystal silicon, stoichiometric and low stress silicon nitride, doped and undoped polysilicon, as well as Arg-Gly-Asp (RGD) peptide adsorbed surfaces characterized in terms of protein adsorption or cellular adhesion for a period of four days. Protein adsorption studies using fibrinogen and albumin, two proteins implicated in cellular adhesion and surface activity, reveal that low stress silicon nitride surfaces have a 223%+/-2.50% greater protein adsorption compared to undoped polysilicon surfaces, followed by silicon nitride, unmodified silicon, and doped polysilicon surfaces, respectively. The thickness of the adsorbed albumin and fibrinogen layer on various thin films was measured by ellipsometry and compared to contact angle measurements. The greatest cellular adhesion was observed on undoped polysilicon, followed by unmodified (control) silicon, low stress silicon nitride, silicon nitride, and doped polysilicon surfaces. Cellular binding supports the differential protein adsorption found on modified and unmodified silicon surfaces. Understanding the biological response to thin films will allow us to design more appropriate interfaces for implantable diagnostic and therapeutic silicon-based microdevices. PMID- 15348210 TI - Hydrolytic and enzymatic degradations of physically crosslinked hydrogels prepared from PLA/PEO/PLA triblock copolymers. AB - Degradable copolymers were synthesized by ring opening polymerization of lactide in the presence of poly(ethylene glycol) (PEG), using CaH2 as a biocompatible initiator. The resulting PLA/PEO/PLA triblock copolymers were dissolved in a biocompatible solvent, namely tetraglycol. Physically crosslinked hydrogels were then prepared by introducing small amounts of water into the thus obtained solutions. Hydrolytic degradation of the highly swollen hydrogels was realized in 0.13 M pH=7.4 phosphate buffer, while the enzymatic degradation was carried out in 0.05 M pH=8.6 Tris buffer containing a PLA-degrading enzyme, proteinase K. In both cases, degradation was initially very fast with dramatic weight loss. The LA/EO ratio of the remaining material increased rapidly, in agreement with the release of PEO-rich segments. In a second phase, the degradation rate slowed down. The presence of proteinase K strongly accelerated the degradation rate of the hydrogels, indicating that the enzyme was able to penetrate inside and attack the PLA domains which constituted nanometric nodes in the gel network. PMID- 15348211 TI - Hydroxypatite-polysaccharide granules for drug delivery. AB - The formation of hydroxyapatite by co-precipitation from sodium alginate and sodium carboxymethylcellulose aqueous solutions with the use of dibasic calcium phosphate dihydrate and calcium hydroxide as starting reagents is studied. A technique to prepare the hydroxyapatite/polysaccharide (micro)granules is developed. An introduction of an antimicrobial Biocide 1 agent in proper amount into the granules is provided, and the behavior of the granules is evaluated. PMID- 15348212 TI - Reactivity of biological and synthetic hydroxyapatite towards Zn(II) ion, solid liquid investigations. AB - The reaction of biological and synthetic hydroxyapatite Ca5(PO4)3OH (HAP) with Zn2+ ions is investigated as a function of Zn2+/Ca2+ molar ratio, time, temperature and electrolyte type (NaCl, NaHCO3, Na2HPO4) by means of pH, pZn, pCa measurements, in aqueous solution. Biological powdered HAP invariably affords an almost quantitative reaction, while Zn2+ precipitated only partially by reaction with cubelets of biological HAP. Using powdered biological HAP and synthetic HAP (dried at 100 degrees C), the reaction with Zn2+ ion is fast and takes place without addition of precipitating anion; synthetic HAP (dried at 1000 degrees C) reacts if free phosphate ions are present. The solid phases separated after different reaction times are investigated by means of X-ray diffraction (XRD), IR, SEM techniques and elemental analysis (C,H,N). The solid phases contain Zn3(PO4)2.4H2O (Hopeite) at the beginning of reaction and CaZn2(PO4)2.2H2O (Scholzite) at the equilibrium. PMID- 15348213 TI - The effects of bovine trabecular bone matrix particulates on cortical bone repair. AB - This paper reports the effects of a synthetic bone substitute and bone allograft on cortical bone repair in an experimental model. To test the hypothesis that bovine trabecular bone matrix, BBM, can enhance the repair rate of cortical bone, osteotomies were created in the rabbit fibula and filled with either allograft or BBM particulates or left empty as controls. At five weeks post-surgery, mechanical tests and histological evaluations were performed. No significant differences were observed in the mechanical properties of the healing bone in the three animal groups (n=6). Histologically, the medullary cavity was obstructed and the cross-sectional area ratio of the osteotomies to intact bone was approximately 3 : 1. Highly significant area differences were observed between the intact bone group and both the BBM and the allograft groups (p< 0.001). At the junction between the original bone and the newly formed bone, both woven and lamellar bone microstructures were prevalent. However, in the BBM filled defects, the woven bone microstructure was not ostentatious. It is concluded that failure to demonstrate significantly differences between the treatments were due to the small sample sizes and or the efficacy of the tensile analysis. PMID- 15348214 TI - Evaluation of biocompatibility of the copolymer of 2-hydroxyethyl methacrylate with 2-(methylsulfanyl)ethyl methacrylate. AB - This study compares subcutaneous and intracerebral biocompatibilty of two hydrogels: copolymer of 2-hydroxyethyl methacrylate with 2-(methylsulfanyl)ethyl methacrylate and poly(2-hydroxyethyl methacrylate) as reference polymer. The experimental copolymer was more biologically inert than poly(2-hydroxyethyl methacrylate) in both the studied parameters, hence the former material is a suitable candidate for biomedical application. PMID- 15348215 TI - Dextran-magnetite complex: temperature dependence of its NMR relaxivity. AB - The NMR relaxivity of proton by dextran-magnetite (DM) complex was examined at temperatures from 20 to 80 degrees C. The decrease in T1 relaxivity with increasing temperature is due to the thermal activation of motion of water molecules in and/or surrounding DM particle. The decrease in magnetization of magnetite core is the secondary effect. The temperature dependence of T1 relaxivity was found to depend on molecular weight of dextran. The motion of dextran chains binding to core may affect the thermal activation of "heavy water" interacted with dextran. The Arrhenius plots of T1 relaxivity showed different slopes at 20-40 degrees C and 50-80 degrees C, while, no change in slope was observed for Fe(NO3)3, CuSO4 and Gd-DTPA solutions. PMID- 15348216 TI - Biocompatibility of poly(vinyl alcohol)-hyaluronic acid and poly(vinyl alcohol) gellan membranes crosslinked by glutaraldehyde vapors. AB - Glutaraldehyde (GTA) solutions are commonly used to crosslink biomolecules and artificial polymers in order to reduce the degradation rate and to avoid the rapid dissolution in biological fluids. The toxicity of these materials is often due to the presence of GTA residuals unremoved by washing procedures. In this study membranes of PVA-hyaluronic acid and PVA-gellan with different composition have been obtained by solution casting technique and crosslinked by exposure to vapors of GTA in acid environment. The harmful effects of GTA residuals released from the membranes have been evaluated by the cytotoxicity and cytocompatibility in vitro tests, based on the cell culture method. The results showed that these materials have no toxic effects: they do not affect cell viability and proliferation, nor exert damages on mithocondrial and lysosomal functions. The poor adhesion of cells seeded directly onto membranes is due to the surface properties of these materials which are completely refractory at cell adhesion and proliferation. The use of GTA in vapor phase as crosslinking agent of natural and artificial polymer blends is demonstrated to be an efficacious procedure that avoids the presence of toxic residuals into materials. PMID- 15348217 TI - Study of a hydraulic calcium phosphate cement for dental applications. AB - Calcium phosphate-based cements (CPCs) have attracted much interest because of their good osteoconductivity for bone reconstruction. We obtained CPCs by mixing calcium bis-dihydrogenophosphate monohydrate (MCPM) and calcium oxide with water or sodium phosphate buffers (NaP) as liquid phase. Cement samples with different calcium-to-phosphate ratios (Ca/P), liquid-to-powder ratios (L/P) and liquid phases were analyzed by X-rays diffraction (XRD), pH-metry, extensometry and calorimetry. Antibacterial activity on two bacterial strains (Streptococcus mutans, Lactobacillus acidophilus) and a polycontaminated bacterial inoculum was also studied using the agar diffusion method. The best mechanical properties (approximately 25 MPa) corresponded to Ca/P ratios between 1.67 and 2.5, a 1 M sodium phosphate buffer pH 7, as liquid phase and a L/P ratio of 0.6ml g(-1). The final setting time increased with the Ca/P ratio. The setting expansion, around 1 2%, depended on the Ca/P and L/P ratios. The inner temperature of the cements rose to 45 degrees during setting then decreased rapidly. The injectability was 100% up to 3.5 min and then decreased. It increased with increasing the L/P ratio but to the detriment of the compressive strength and setting time. XRD analysis indicated that the setting reaction led to a mixture of calcium hydroxide and calcium-deficient hydroxyapatite even for a Ca/P ratio of 1.67. Consequently, the pH of the surrounding fluids rose to 11.5-12 during their dissolution. Bacterial growth inhibition was only clearly observed for Ca/P>or=2. This bioactive calcium phosphate cement can potentially be employed for pulp capping and cavity lining as classical calcium hydroxide-based cements, but it is not usable, in the present formulation, for root canal filling because of its short setting time. PMID- 15348219 TI - Materials science: materials in medicine. PMID- 15348218 TI - Effect of poling conditions on growth of calcium phosphate crystal in ferroelectric BaTiO3 ceramics. AB - Recently, ceramic materials have been given a lot of attention as candidates for implant materials, since they possess biologically favorable characteristics for osseointegration. Among them, BaTiO3 (BTO) ceramics are ferroelectric and piezoelectric after poling treatments. However, little or no information is available on the poling condition of BTO and their effect on calcium phosphate (CaP) formation. In this study, the effect of poling conditions on the formation of CaP layer was investigated. It was observed from this study that CaP was formed on negatively charged BTO surfaces. An increase in Ca/P ratio to 1.67 was observed when the poling temperature was increased above the Curie temperature. On positively charged BTO, no CaP layer was observed. PMID- 15348220 TI - pH dependent hydrolysis and drug release behavior of chitosan/poly(ethylene glycol) polymer network microspheres. AB - Semi-interpenetrating polymer network (IPN) microspheres of chitosan and poly(ethylene glycol) PEG were prepared for controlled release of drugs. A new method for the chemical crosslinking of chitosan microspheres containing isoniazid (INH) as a model drug is proposed and evaluated. The method consists of the exposure of microspheres to the vapor of crosslinking agent that act in gaseous phase under mild conditions. The structural analysis of the microspheres was carried out by FTIR-analysis. The swelling behavior, hydrolytic degradation, structural changes of the microspheres and loading capacity (LC) of the microspheres for INH were investigated. The prepared microspheres have shown 93% drug loading capacity, which suggested that these semi-IPN microspheres are suitable for controlled release of drugs in an oral sustained delivery system. PMID- 15348221 TI - Use of an osteoinductive biomaterial as a bone morphogenetic protein carrier. AB - A porous calcium phosphate ceramic, which induced bone formation in soft tissues of dogs, was termed as osteoinductive biomaterial and studied as a carrier of bone morphogenetic protein (rhBMP-2). Cylinder implants (laser spot 4 x 5 mm) impregnated with 0, 1, 10 and 40 microg rhBMP-2 were implanted in dorsal muscles of rabbits for five weeks. Histological observation and histomorphometric analysis were performed on thin un-decalcified sections. No bone formation was detected in the implants without rhBMP-2, while mature lamellar bone was found inside the implants with 1 microg rhBMP-2, both on the outer surface and inside the implants with 10 microg and 40 microg rhBMP-2. Little difference in formed bone was found between 1 microg and 10 microg rhBMP-2, but no difference was found between 10 microg and 40 microg rhBMP-2. A significant difference in bone marrow formation was found among 1, 10 and 40 microg rhBMP-2. The more rhBMP-2, the more bone marrow formed. The present results indicate that osteoinductive biomaterial is a good carrier of BMP and high dose of BMP is not necessary for bone formation in clinic. PMID- 15348222 TI - Crystallization kinetics of sol-gel derived hydroxyapatite thin films. AB - The crystallization kinetics of sol-gel derived hydroxyapatite (HA) and tricalcium phosphate (TCP) thin films were studied to determine whether viscous sintering could be used for densification. The films were approximately 900 nm thick, and were synthesized and processed on silicon substrates. The films were fired in air in a rapid thermal annealer (RTA) for various times and the degree of crystallinity was determined by measuring the intensity of characteristic X ray diffraction lines. The growth kinetics of HA and TCP were measured between 420 and 550 degrees C, and between 840 and 920 degrees C, respectively. Films that were subjected to an accelerated aging step before firing, exhibited a significantly lower crystallization growth rate when compared to unaged films. The aged films also became harder, as measured by nanoindentation. At temperatures above 840 degrees C, HA transformed into both alpha-and beta-TCP, with the beta form being dominant at lower temperatures. The activation energies for both transformations (amorphous film to HA, and HA to TCP) were determined, as were the constants for the Avrami equation. Based on the rapid crystallization kinetics observed for the amorphous film to HA transformation, densification through viscous sintering is essentially precluded in this system. PMID- 15348223 TI - Bone response to polymers based on poly-lactic acid and having different degradation times. AB - Authors studied two degradable and resorbable polymers derived from lactic acid: poly-L-Lactic acid (PLLA), with a relatively long time of degradation (longer than 6 months, PL10 Purac NL); poly-DL-Lactic acid (PDLLA), with a relatively short time of degradation (shorter than 6 months, PDL Purac NL). The animal species was the young adult New Zealand White rabbit. The in-vivo study was performed by implantation of small cylinders of 10 x 3 mm in size (length x diameter) in the distal metaepiphysis of the femur; 34 cylinders have been implanted. Retrievals of PLLA specimens took place at 3, 6, 9, 12 and 24 months; for PDLLA specimens at 1, 2, 4 months. Polarized light microscopy of undecalcified tissue sections was performed. The analysis for PLLA and PDLLA has shown a favorable response of bone tissue: alterations in the bone repair, growth and remodeling have not been observed. PLLA is persistent at the times studied; there is never a tight apposition between bone and PLLA implant and an intervening fibrous layer has often been observed. PDLLA is not persistent at the times studied and it degrades quite fast; bone repair of the empty implantation's hole occurs by bony growth from the endosteal trabeculae. The newly formed bone covers the hole's walls with an elongation parallel to them. For both polymers, whether the degradation is fast or slow, the material's substitution by newly formed bone never starts from the walls of the implantation hole. Only after the complete disappearance of the polymeric material newly formed bone begins to fill the hole. PMID- 15348224 TI - Diffuse damage accumulation in the fracture process zone of human cortical bone specimens and its influence on fracture toughness. AB - This study was concerned with the mechanics and micromechanisms of diffuse (ultrastructural) damage occurrence in human tibial cortical bone specimens subjected to tension-tension fatigue. A nondestructive technique was developed for damage assessment on the surfaces of intact compact tension specimens using laser scanning confocal microscopy. Results indicated that diffuse damage initiates as a result of fractures in the inter-canalicular regions. Subsequent growth of those microscopic flaws demonstrated multiple deflections from their paths due to 3D spatial distribution of microscopic porosities (lacunae canalicular porosities) and the stress-concentrating effects of lacunae. Damage dominating effects in the early stages of fatigue had been verified by the observed variations of the fracture toughness due to artificially induced amounts of damage. Toughening behavior was observed as a function of diffuse damage. PMID- 15348225 TI - Mineralization of regenerated cellulose hydrogels. AB - Due to their high water swelling, regenerated cellulose hydrogels and sponges were pre-incubated in a Ca-containing solution, and their mineralization was investigated. Results obtained demonstrate that a simple pre-incubation treatment in a Ca containing solution can induce mineralization in materials with limited or no tendency to mineralize. The minerals formed had an apatitic carbonated and poorly crystalline structure, resembling carbonated hydroxyapatite found in bone mineral. The apatitic layer formed showed a relatively accelerated growth using this technique, exhibiting nodules in their macroscopic structure, which seem to indicate lateral growth. The porous structure of regenerated cellulose sponges was also homogeneously mineralized using this technique. PMID- 15348226 TI - Synthesis and evaluation of various layered octacalcium phosphates by wet chemical processing. AB - Octacalcium phosphate was synthesized by hydrolysis of alpha-tricalcium phosphate through a wet-chemical processing. Using the same wet-chemical processing in presence of various succinate ions, the preparation of some complexed octacalcium phosphates was attempted. The products were examined by X-ray diffraction method. These complexed octacalcium phosphates intercalated with succinic acid, L asparatic acid, and methyl succinic acid showed an expanded basal spacing in the octacalcium phosphate unit cell dimensions. The microstructure was observed by scanning and transmission electron microscopy. PMID- 15348227 TI - Release of sodium fluoride from poly (L-lactic acid) implants characterized by thermal history. AB - The method of injection molding of the melt of poly(L-lactic (PLLA), Mw 11 730, containing 7.5% of particles of sodium fluoride smaller than 1 microm was employed to prepare cylinders of a diameter of 2 and 10 mm in length. These matrices with an insoluble active ingredient with an amorphous structure of the carrier obtained by rapid cooling of the melt were annealed at temperatures from 70 to 140 degrees C in the medium of a hot-air drying plant and liquid paraffin. It has been found that crystallization of PLLA manifested by the rapidity of release of sodium fluoride does not take place at a temperature of 70 degrees C; at a temperature of 75 degrees C the effect of annealing of polyester matrices on the decrease in the rapidity of sodium fluoride release was already perceptible. Also in annealing of matrices at temperatures of 120 and 140 degrees C an identical deceleration of sodium fluoride release as that at 75 degrees C took place; the rapidity of the development of an effective crystalline phase was higher at higher temperatures. The method of interval cooling with the first stage of slow cooling and the second stage of rapid cooling of the melt of PLLA with 15% sodium fluoride fixed the structure achieved till the moment of the change in the rapidity of cooling. This revealed the temperature range of the development of the effective crystalline phase from the viewpoint of sodium fluoride release lying between 92 and 122 degrees C. PMID- 15348228 TI - Immobilization of the thrombin inhibitor r-hirudin conserving its biological activity. AB - Surface immobilization of the thrombin inhibitor r-hirudin was carried out on two different polymers. Linkage to poly(urethane-graft-acrylic acid) (PAC/PU) was done via carboxylic acid groups, using a water soluble carbodimide, while the immobilization on a modified poly[(ethene-co-vinyl acetate)-graft-vinyl chloride] (PVC/EVA) was achieved via the alcohol groups of the polymer using HDI as spacer. Direct immobilization of r-hirudin leaded to a remarkable loss of thrombin activity. As proved by means of protein chemical analysis, loss of activity was due to a selective coupling via the N-terminal amino group of r-hirudin, which is essential for its thrombin activity. Based on these results we developed an immobilization method via an epsilon-amino group of r-hirudin preserving full biological activity of the r-hirudin coated surface. PMID- 15348229 TI - Polyglycolide: degradation and drug release. Part I: changes in morphology during degradation. AB - The changing morphology of quenched polyglycolide (PGA) is investigated during hydrolytic degradation in phosphate buffered saline at pH 7.4. Analysis techniques include small and wide-angle X-ray scattering (SAXS and WAXS), mass measurements, DSC, pH measurement and UV-spectrophotometry. It is postulated that the degradation process can be separated into four distinct stages. In stage I, water diffuses quickly into the sample. During stage II, the polymer crystallizes by insertion crystallization, whilst the molecular weight gradually falls. This stage is characterized by a dramatic fall in the long period together with an increase in the crystallinity, minimal mass loss and minimal water uptake. At the onset of stage III, at around 10 days, a critical molecular weight is reached. Degradation products are now small enough to diffuse from the surface of the sample which begins to swell, water diffuses into the space created, and the crystals are freed from constraint. A co-operation between degradation products diffusing out of the sample and the water diffusing in causes "reaction-erosion" fronts to develop inside the sample. Ahead of these fronts, the trapped acidic degradation products remain to catalyze the hydrolysis. Stage III is characterized by swelling and an increase in the long period, together with mass loss and further water uptake. It is postulated that these reaction-erosion fronts move through the sample and meet in the centre at the beginning of stage IV, at which point the degradation again becomes homogeneous throughout the sample. PMID- 15348230 TI - Polyglycolide: degradation and drug release. Part II: drug release. AB - This paper considers drug release from a polyglycolide (PGA) matrix and is divided into two sections. The first investigates the effects on the degradation of the polymer of incorporating a model drug, theophylline, into the polymer. Small and wide angle X-ray scattering, and mass loss and water uptake measurements indicate that the presence of this drug does not affect the time scale of the degradation process. However, the dissolved theophylline molecules affect the extent to which the polymer crystallizes during degradation. In the second section, theophylline release profiles, obtained using UV spectrophotometry, show that the erosion of the polymer controls the release of the drug. The drug release results add further evidence to support the four stage degradation process which was described in Part I. PMID- 15348231 TI - Production and evaluation of hydroxyapatite reinforced polysulfone for tissue replacement. AB - A variety of bioactive composites have been developed for tissue replacement over the last two decades. In this investigation, a new material consisting of hydroxyapatite (HA) and polysulfone (PSU) was produced and evaluated for potential medical applications. The HA/PSU composite containing up to 20 vol % of HA was studied at the initial stage. It was manufactured via a standardized procedure which included drying, blending, compounding and injection/compression molding. Defect-free composite samples (rectangular bars, discs and dumbbell specimens) could be obtained by injection molding. Thick composite plates could be made by compression molding. Both compounded materials and molded parts were assessed using a variety of techniques. It was found through scanning electron microscopy (SEM) that HA particles were well dispersed in the PSU matrix. Thermogravimetric analysis (TGA) verified the amount of HA in the composite. Differential scanning calorimetry (DSC) results indicated that the glass transition temperature (Tg) of the polymer matrix was not affected by the incorporation of HA. Rheological analysis revealed that PSU and the composite exhibited pseudoplastic behavior. For unfilled PSU, its viscosity decreased with an increase in temperature. The viscosity of HA/PSU composite increased with an increase in the HA volume fraction. It was shown through dynamic mechanical analysis (DMA) that the storage modulus of the composite was increased with an increase in HA volume percentage below Tg of the polymer, while tan delta was maintained at nearly the same level. It was established that water uptake reached an equilibrium after 7 days' immersion in distilled water for PSU and HA/PSU composite. After 7 days' immersion in distilled water, the storage modulus of the composite was decreased less than that of PSU. PMID- 15348232 TI - Surface characterization and platelet adhesion studies on polyethylene surface with hirudin immobilization. AB - Hirudin, a protein composed of 65 or 66 amino acid, is a newly risen anticoagulant agent and has been considered as the most potent thrombin inhibitor. Hirudin can block the active site of thrombin by means of its carboxylic acid reaction with the active regime of thrombin, and becomes a tightly bound complex, and thus controls the formation of thrombus. Hirudin was covalently immobilized onto the water-soluble carbodiimide preactivated and chromic acid oxidized PE surface. Surface chemistry analysis indicated that a certain amount of carboxylic acid groups was generated on the polyethylene surface after oxidation with chromic acid solution. The amount of carboxylic acid functional group increased with the oxidation time. In addition, polyethylene surface was etched by chromic acid solution, and a rougher surface was created. The morphology of oxidized polyethylene surface was similar to each other among the samples with oxidation time from 1 to 8 min. ESCA results indicated the number of hirudin molecules immobilized was constant at the reaction time studied. In vitro platelet adhesion assay indicated the number of adhered platelets on the oxidized polyethylene surface increased significantly after oxidation. In contrast, surface with hirudin immobilization showed a reduction in adhered platelet density than the chromic acid oxidized counterpart due to the decrease of platelet-activating capability by the hirudin-thrombin complex and the differences in the adsorbed protein composition. PMID- 15348233 TI - The influence of crosslinking agents and diamines on the pore size, morphology and the biological stability of collagen sponges and their effect on cell penetration through the sponge matrix. AB - Artificial skin substitutes based on autologous keratinocytes cultured on collagen substrata are being developed for treating patients with severe burns. The properties of the collagen substrate can be manipulated, for example, by crosslinking, to optimize desirable properties such as cell growth and penetration into the substrate, biological stability and mechanical strength. Collagen sponges crosslinked with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC) and the diamine, diaminohexane, were used to determine the effect of crosslinking on pore size and morphology, on the stability of the crosslinked sponges both in cell culture media and during incubation with collagenase, and on the penetration of keratinocytes and fibroblasts through the sponge matrix. Crosslinking of the sponges reduced the pore size, particularly at the surface, and altered sponge morphology. After crosslinking the collagen fibers were thinner, and appeared lacy and delicate. Crosslinking also influenced sponge stability. In keratinocyte serum-free medium the pore size of plain collagen sponges increased with increasing incubation time, and crosslinking appeared to prevent this, and may have stabilized sponge structure. Incubation in serum containing Dulbecco's minimum essential medium caused a marked reduction in pore size in both plain collagen and crosslinked collagen sponges. Crosslinking did not appear to influence this cell-free contraction of collagen sponges. Treatment of sponges with EDAC markedly increased the resistance of sponges to collagenase digestion. The penetration of both keratinocytes and fibroblasts was retarded by crosslinking the sponges. Fibroblasts penetrated through the sponges to a greater extent than keratinocytes, and their proliferation rate was faster. The total number of cells populating the crosslinked sponges after 10 days culture was approximately 50% of that on untreated collagen sponges. The mechanism responsible for this effect was different with the two crosslinkers used. Diaminohexane appeared to inhibit cell growth, whereas EDAC may have caused a decrease in cell adhesion to the sponges, without an apparent inhibition of growth rate. In terms of morphology, fibroblasts were elongated to a greater extent on crosslinked sponges, and alligned themselves along the collagen fibers. Keratinocytes grew in colonies on untreated sponges, but on crosslinked sponges they grew in isolation, with minimal cell-cell interactions. It may be necessary to reach a compromise to obtain the best combination of properties for using collagen sponges as substrata for artificial skin substitutes. PMID- 15348234 TI - Hydroxyapatite particles are capable of inducing osteoclast formation. AB - Hydroxyapatite (HA) coatings have been used to improve implant fixation by promoting bone formation around the prosthesis. A macrophage response to HA particulates has been noted around loosened HA-coated prostheses. As biomaterial wear particle-associated macrophages are known to be capable of differentiating into osteoclasts that are capable of bone resorption, we examined whether particulate HA could similarly induce macrophage-osteoclast differentiation. HA associated macrophages were isolated from granulomas, formed by subcutaneous implantation of HA, and co-cultured with UMR 106 osteoblast-like cells in the presence of 1,25-dihydroxyvitamin D(3) for up to 14 days on glass coverslips and bone slices. HA-associated macrophage-osteoclast differentiation was evidenced by the formation of numerous multinucleated tartrate resistant acid phosphatase (TRAP)-positive cells which formed lacunar resorption pits on bone slices. Polymethylmethacrylate (PMMA) particle-associated macrophages, isolated from subcutaneous PMMA-containing granulomas, caused significantly more osteoclast formation and bone resorption than HA-associated macrophages. These results indicate that macrophages responding to HA particles are capable of osteoclast differentiation. They also suggest that particles derived from uncemented (HA coated) implants are likely to induce less osteoclast formation and osteolysis than cemented implants. PMID- 15348235 TI - A new method for selecting calf pericardium for use in cardiac bioprostheses on the basis of morphological and mechanical criteria. AB - The durability of existing calf pericardium bioprostheses is limited by phenomena such as mechanical stress and calcification, the factors most frequently implicated in valve failure. Varying the preferred direction of the collagen fibers influences the mechanical behavior of the pericardial membrane. Given this possible variation, a strict control of the selection of the biomaterial employed in the construction of valve leaflets is essential, but a reliable method of selection has yet to be established. This study describes the development of a new system of in vitro selection involving a hydraulic simulator that reproduces the mechanical behavior of pericardial membranes subjected to the stress of continuous flow. By combining morphological criteria such as thickness and homogeneity with those of mechanical behavior, and by selecting paired samples from different parts of the pericardium, we obtained excellent mathematical fits. Linear regression analysis provided the mode of predicting the tensile strength in a given sample when this value had been determined in its twin. The upper zones of calf pericardium, corresponding to either right or left ventricle but at a distance from ligamentous structures, showed the best mean results at rupture (60 MPa) and permitted the most reliable prediction. The expected stress for an elongation of 30% was 1.12 MPa, as was previously observed, with a 95% confidence interval of between 1.11 and 1.14 MPa. These trials, together with the careful selection of the pairs, should help to establish definitive selection criteria. PMID- 15348236 TI - The influence of polymer blend composition on the degradation of polymer/hydroxyapatite biomaterials. AB - The in vitro degradation of biodegradable polymer/ceramic composites was assessed in two different environments under both static and pseudodynamic conditions. The blends, consisting of polycaprolactone, poly(lactic-co-glycolic acid), and hydroxyapatite, have potential use in bone tissue engineering applications, thus it is essential to establish a standardized method of characterizing the degradation of new biomaterials. In this study, the variation in polymer blend ratio was examined to observe a change in degradation rate. The porous blends were degraded in water and serum-containing media. A previous study examined in vitro degradation in serum-free buffer. Molecular weight loss, gravimetric weight loss, pH changes and morphological changes were evaluated. The changes in porosity were observed with scanning electron microscopy and quantitatively assessed using image analysis. There was a significant difference in molecular weight loss and gravimetric weight loss between the blends after 10 weeks in vitro. Blends containing the greatest amount of poly(lactic-co-glycolic acid) degraded most rapidly. PMID- 15348237 TI - Porous HA ceramic for bone replacement: role of the pores and interconnections - experimental study in the rabbit. AB - Hydroxyapatite (HA) porous ceramics are increasingly used in biomedical applications. Their physical characteristics, such as porous volume, require perfect control of the pore shape, as well as the number and the size of their interconnections. The aim of our study was to evaluate a new HA ceramic using polymethylmethacrylate microbeads (PMMA) as the porous agent. Four interconnection sizes (30, 60, 100 and 130 microm) with a 175-260 microm pore size and three pore sizes (175-260, 260-350 and 350-435 microm) for a 130 microm interconnection size were tested. Various HA implants were appraised by microscopic evaluation in a 4.6 x 10 mm rabbit femur cancellous bone defect 12 weeks after implantation. The best osteoconduction result was obtained in the center of the ceramic by means of a 130 microm interconnection size and a 175-260 microm mean pore size. Bone formation obtained within the pores was double that obtained in our previous study where naphtalen microbeads were used as the porous agents. PMID- 15348238 TI - Effect of thermal treatment on apatite-forming ability of NaOH-treated tantalum metal. AB - The prerequisite for an artificial material to bond to living bone is the formation of bonelike apatite on its surface in the body. This apatite can be reproduced on its surface even in an acellular simulated body fluid with ion concentrations nearly equal to those of the human blood plasma. The present authors previously showed that the tantalum metal subjected to a NaOH treatment to form a sodium tantalate hydrogel layer on its surface forms the bonelike apatite on its surface in SBF in a short period. The gel layer as-formed on the metal is, however, not resistant against abrasion, and hence thus-treated metal is not useful for clinical applications. In the present study, effects of thermal treatment on the mechanical properties and apatite-forming ability of the NaOH treated tantalum metal were investigated. The sodium tantalate gel on the NaOH treated tantalum was dehydrated to convert into amorphous sodium tantalate by a thermal treatment at 300 degrees C in air environment and into crystalline sodium tantalates by the thermal treatment at 500 degrees C. Resistivity of the gel layer against both peeling-off and scratching was significantly improved by the thermal treatment at 300 degrees C. The high apatite-forming ability of the sodium tantalate hydrogel was a little decreased by the thermal treatment at 300 degrees C, but appreciably decreased by the thermal treatment at 500 degrees C. It is believed that the tantalum metal subjected to the 0.5 M-NaOH treatment and the subsequent thermal treatment at 300 degrees C is useful as implants in dental and orthopaedic fields, since it shows high bioactivity as well as high fracture toughness. PMID- 15348239 TI - Viscoelastic properties of the nucleus pulposus of the intervertebral disk in compression. AB - The viscoelastic properties of the nucleus pulposus were measured in compression for 35 specimens dissected from 9 sheep. Measurements on 19 specimens were made on the day of slaughter; the remaining 16 specimens were stored frozen and thawed before testing. A preload of 0.2 N was applied to each specimen and a cyclic compression of 10 microm applied at eight frequencies in the range 0.1-10 Hz at a temperature of 37 degrees C. Freezing appeared to increase the storage modulus, E ', but not the loss modulus, E ", or tan delta=E "/E '. These parameters, E ', E " and tan delta, had values of 64+/-28 kPa, 24+/-11 and 0.33 kPa+/-0.07, respectively. The value of tan delta passed through a minimum at a loading frequency of 0.9+/-0.2 Hz. The water content of the specimens was 80+/-2%. PMID- 15348240 TI - Biochemical and histological evaluation of human synovial-like membrane around failed total hip replacement prostheses during in vitro mechanical loading. AB - The biochemical role of the synovial-like membrane formed at the interface of eight aseptic failed total hip prosthesis has been investigated during in vitro mechanical loading. The study was carried out on four membranes from cemented prosthesis and four titanium alloy uncemented ones. Intermittent positive pressure leading to 20% deformation of the membrane (100 g/cm(2))was applied to the membrane fragments in cycles (300 cycles/15 min) repeated three times at thirty minutes intervals in which interleukin-6 (IL6), prostaglandin-E2 (PGE2) and interleukin-1beta (IL1beta) levels were quantified both in culture media and in tissue extracts. Histological, morphometrical and immunohistochemical studies were also carried out on the same membranes. Mechanical stress evidenced an increase in the release of the examined cytokines both in cemented and uncemented prosthesis tissues; particularly evident was IL6 trend of increase from cemented prosthesis and IL1beta result from uncemented ones. Histomorphological and immunohistochemical data revealed no differences between membranes obtained from cemented and uncemented prosthesis as to cell proliferation, fibrosis, macrophages lymphocytes B and T population, vessels and nervous fibers. The results indicate that mechanical stress plays a fundamental role in increasing membrane production and release of cytokines known as bone-resorbing agents. Furthermore, the histologic finding of synovial-like membrane with the same histomorphological and immunohistochemical findings but with different biochemical response to mechanical stimulation, suggests that cells involved in the production and release of the considered mediators might have different strain behavior by different development conditions (previous contact with PMMA). PMID- 15348241 TI - The distribution of osteocalcin, degree of mineralization, and mechanical properties along the length of Cyprinus carpio rib bone. AB - This study examined the spatial distribution of selected biochemical and mechanical properties along the length of carp rib bone. Carp rib bone was chosen because of its unusually high osteocalcin content relative to other extractable proteins. The amount of osteocalcin was significantly lower (p<0.01) at the most distal section, relative to all other sections. The amount of phosphate (p<0.05) and the elastic modulus in the longitudinal plane (p<0.0001) were found to be significantly higher in the most distal section, relative to the most proximal section. There was no significant difference in the calcium distribution, molar Ca/P ratio, or elastic modulus in the transverse plane. It was speculated that the distal section contains less mature bone. The methods illustrate the potential usefulness of nanoindentation to characterize the mechanical properties of bone, relative to its biochemical composition. PMID- 15348242 TI - Control of hydroxyapatite crystallinity by mechanical grinding method. AB - Crystallinity of hydroxyapatite reflecting crystal size and crystal elastic strain was controlled by the mechanical grinding (MG) technique using a set of container and balls made of SUS304 stainless steel or agate. Variation in the crystallinity through MG was monitored by the XRD method and represented by the broadening of the diffraction peak. Effect of changes in crystallite size and strain on the crystallinity was also examined using the Hall-plot method. Crystallinity rapidly decreased with milling time. Significant crystallographic diffraction peaks disappeared and a broad diffraction around 2theta=32 degrees was observed after MG for 72 h. The broadening was dominantly due to an increase in crystal strain in addition to fine crystallite size. Contamination from the container and balls during MG was more suppressed using agate than SUS304 stainless steel. The recovery process of crystallinity during heating between 300 degrees C and 1200 degrees C was examined focusing on the decrease in residual elastic strain. Low crystallinity was maintained at annealing temperatures below 800 degrees C but lattice defects were recovered above 1000 degrees C. PMID- 15348243 TI - Effects of environment on the creep properties of a poly(ethylmethacrylate) based bone cement. AB - The effect of test environment on the creep behavior of a poly(ethylmethacrylate) bone cement was investigated. The aim of the study was to assess the influence of environment on the inherent material behavior, and so it was convenient to perform tests in tension on well-prepared samples. In addition to control tests in air, the liquid environments studied were water, Ringer's solution and Intralipid. Creep tests were performed in each of these environments with a range of aging times, test temperatures and applied stresses. In order to compare the effects of the environments, the creep curves were fitted to a generalized form, from which a creep rate was determined. The ratio of these creep rates between different environments at each testing condition was then used as a basis for a comparison of the detailed effects of environment. It was found that in all cases the water-based environments (water and Ringer's) had similar effects and gave the largest creep rates. Intralipid was then intermediate and air gave the lowest creep rates. These effects are mainly due to plasticization by water, although with Intralipid, some increased monomer leaching occurred, which served to reduce the creep rates. The influence of environment on the effects of aging time, temperature and stress were complex, although in general any conditions which increased water plasticization (longer aging, higher temperature and to a lesser extent, higher stress) gave an increase in creep rate. The major exception to this was at temperatures of 40 degrees C and above, where the effects of water plasticization were diminished, due to the inherent increase in molecular mobility of the material. PMID- 15348244 TI - Mechanical and structural characteristics of commercially pure grade 2 Ti welds and solder joints. AB - This study aimed at determining whether data previously gathered for a laser welds and IR brazings using a Au-Pd alloy were applicable to titanium joints. As to its resistance under fatigue loading, Au-Pd alloy had shown a poor response to pre-ceramic laser welding and post-ceramic brazing. The present study was designed to assess the mechanical resistance, the microstructure and the elemental diffusion of laser welded, electric arch welded and brazed joints using commercially pure titanium as substrate metal. Mechanical resistance was determined by determining the joints' ultimate tensile strength and their resistance to fatigue loading. Elemental diffusion to and from the joints was assessed using microprobe tracings. Optical micrographs of the joints were also obtained and evaluated. Under monotonic tensile stress, three groups emerged: (1) the GTAW and the native (i.e. as received) substrate, (2) the annealed substrate and the laser welds and (3) the brazed joints. Under fatigue stress, the order was: first the native and annealed substrate, second the brazings and laser welds, third the GTAW joints. No Au-filler brazing withstood the applied fatigue loading. The micrographs showed various patterns, an absence of HAZ cracking and several occurrences of Widmanstatten structures. Elemental diffusion to and from the Ti substrate was substantial in the Ti filler brazings and virtually nil in the Au-based brazings. Under fatigue stress application, the titanium-based brazings as well as the laser- and electric arc welds performed equally well if not better than a previously tested AuPd alloy. There was a definite increase in grain size with increased heat application. However, no feature of the microstructures observed or the elemental analysis could be correlated with the specimen's resistance to fatigue stress application. PMID- 15348245 TI - Energy dispersive analysis (EDX) of a degradable bioactive-glass coating on Ti6Al4V in-vivo. AB - The bioactive-glass coating of metallic substrates provides a gradually degrading interface which can be used to favor the bony integration of the implant by the physiologic processes of bone turn-over and remodeling. Twelve New Zealand White rabbits, about 2700 g of weight, were operated by the sagittal insertion of a bioactive-glass coated plate of Ti6Al4V. Retrievals were performed at 4, 8 and 12 weeks. Undecalcified specimens were embedded in methyl-metacrylate and sectioned at 100 microns of thickness. Blocks were grinded and had an electroconductive coating to be examined by scanning electron microscopy (SEM), back scattering electron microscopy (BSEM) and X-ray energy dispersive spectroscopy microanalysis (EDX). EDX allows to evaluate quantitatively the gradual process of coating degradation. Areas of 200 microns in square were analyzed at the interface between bone and coating to determine their elemental composition. Silicon was the key marker for the presence of the glass. Morphological analysis confirms that a tight apposition with bone can be obtained by utilizing the bioactive glass coating of metal. Results of energy dispersive analysis support the mechanism of a gradual degradation of the bioactive glass coating and its integration with bone, since the presence of silicon can be documented within the newly formed bone after the coating has disappeared. PMID- 15348246 TI - Surface reaction of stoichiometric and calcium-deficient hydroxyapatite in simulated body fluid. AB - In the present study, the immersion behavior of two kinds of sintered HA with different Ca/P ratios in two different extracellular simulated solutions (Tris buffer and Hank's solutions) was investigated and compared. Results indicated that an as-received Ca-deficient HA (FHA) had a lower Ca/P ratio, larger linear shrinkage and higher density than a stoichiometric HA (MHA). When FHA powder was calcined at 900 degrees C, its Ca-deficient apatite structure was unstable and a significant amount of beta-TCP phase was formed. When heated to 1250 degrees C in air, the highly crystalline apatite structure of MHA was still stable without any noticeable decomposition. The FTIR spectra indicated that both immersed MHA and FHA in Hank's solution were gradually covered with a layer of precipitated apatite during immersion. When immersed in Tris buffer solution, neither HA showed significant changes in their FTIR spectra. SEM observation indicated that the precipitation rate on immersed FHA surface was much higher than that on MHA surface when immersed in Hank's solution. The weight loss and pH data confirmed the higher dissolution rate of FHA than MHA in Hank's solution. PMID- 15348247 TI - The spatial arrangement of tubules in human dentin. AB - We applied two-dimensional numerical methods to describe the spatial arrangement of tubules in human dentin. The methods considered were two-point correlation functions, entropy-like measures, and angular distributions between nearest neighbors. The correlation functions were based on Fourier transform methods. The latter two approaches were based on stochastic geometry, and involved developing the Delaunay tessellations of the tubule patterns and their dual Voronoi diagrams. We discovered that for analyzing the distribution of tubules the geometric methods of lattice tessellations were more sensitive to structural order of the tubules than were Fourier-based schemes. Analysis of the data indicated that dentinal tubules are highly ordered in normal dentin. PMID- 15348248 TI - Hydroxyapatite as a filler for dental composite materials: mechanical properties and in vitro bioactivity of composites. AB - Hydroxyapatite (HAp) powder was treated with gamma-methacryloxypropyltrimethoxy silane (gamma-MPS) using standard techniques in both non-polar and polar systems. Infrared spectra (DRIFT) and thermogravimetric analysis (TGA) confirmed the presence of gamma-MPS on the surface of the HAp filler particles. Series of experimental composites consisting of bisphenol-a-glycidyl methacrylate (BisGMA) based resin and untreated or treated HAp filler were produced to determine the mechanical properties and in vitro bioactivity. The incorporation of HAp filler into the BisGMA base resin had an enhancing effect on the flexural strength and Young's modulus of the base resin, the latter being increased by a factor of three. The mechanical properties of the filled resin were not affected by the surface treatment of the HAp, but filler loading was found to have a significant effect on Young's modulus. Higher proportions of silane-treated HAp of smaller particle size could be incorporated in the monomer phase giving rise to composites of higher stiffness. Examination of the fracture surfaces showed that the silanized HAp particles maintained better contact with the polymer matrix. In vitro study revealed that the composites incorporating silanized HAp formed a compact and continuous calcium phosphate layer on their surface after 4 weeks immersion in a simulated body fluid (SBF). PMID- 15348249 TI - Enhancement of bone ingrowth in a titanium fiber mesh implant by rhBMP-2 and hyaluronic acid. AB - The present study was designed to evaluate the efficacy of high molecular weight hyaluronic acid (HA) used as a carrier of recombinant human bone morphogenetic protein-2 (rhBMP-2) adsorbed to a titanium fiber mesh implant (TFMI) in vivo. The quantity of HA in the TFMI rapidly decreased during the initial 3-day period after implantation. BMP particles were trapped by the meshwork of HA as observed by scanning electron microscopy (SEM). TFMIs containing LF-6, HA, rhBMP-2, or HA combined with rhBMP-2 were implanted on the cranium of rats. Analysis of digitized SEM images of samples obtained six weeks post-implantation was performed to determine the area occupied by new bone. The area fraction of Ca relative to that of the pores of TFMI in the HA group was larger than that in the Ti group (p<0.05). The area fraction of Ca in both the BMP and HA+BMP groups was larger than that in both the Ti and HA groups (p<0.01), and that in the HA+BMP group was larger than that in the BMP group (p<0.05). It is suggested that HA is not only an effective carrier of BMP, but also it may have a positive effect on the generation of new bone in the TFMI. PMID- 15348250 TI - The effects of osteoprotegerin on the mechanical properties of rat bone. AB - Osteoprotegerin (OPG) is a naturally secreted protein that decreases bone resorption by inhibiting osteoclast differentiation and activation while promoting osteoclast apoptosis [8]. In this study, the effects of osteoprotegerin injections on long bone mechanical and material properties were investigated in young male Sprague-Dawley rats. OPG increased fracture strength at the femur mid diaphysis in three-point bending by 30%, without affecting the elastic or maximum strength. At the femoral neck, OPG significantly increased the elastic (45%), maximum (15%), and fracture (35%) strengths. There was not a difference in microhardness at the femur mid-diaphysis in comparing the placebo and OPG groups. There were, however, significant increases in whole bone dry mass (25%), mineral mass (30%), organic mass (17%), and percent mineralization (4%); percent mineralization at the mid-diaphysis (3%); and percent mineralization at the distal epiphysis (6%) due to the OPG treatment. While OPG decreased endocortical bone formation (52%), total bone area, endocortical bone area, and periosteal bone formation were maintained with OPG treatment. A 30% increase in the X-ray opacity of the bone at the proximal metaphysis of the right tibiae was observed. Overall, OPG increased mineralization and strength indices in the rat femur. Its effects on strength were more pronounced in the femoral neck than at the mid diaphysis. PMID- 15348251 TI - A surface modified ODS superalloy by thermal oxidation for potential implant applications. AB - In the present work attention is paid on the composition, structure and protective properties of alumina layer produced by high temperature oxidation on MA 956 superalloy (Fe-20Cr-4.5Al-0.5Ti-0.5Y(2)O(3) (wt %)). The combination of good mechanical properties of this material and the excellent biocompatibility, the good wear and corrosion behavior of an outer alpha-alumina layer, limiting the release of ionic species and wear debris from the bulk material into the body fluid environment, can make this material a candidate alloy for medical applications. Isothermal oxidation at 1100 degrees C in air of the alloy has led to the formation of a fine-grained, compact and adherent alpha-alumina scale. Oxide nodules rich in Ti, Y, Cr, and Fe were found on the top of the surface. In vitro electrochemical corrosion experiments showed good protective properties of the oxide scale. Moreover, no spallation of the alumina layer was observed. This feature is significant considering that the alumina layer has to withstand very high compressive stresses resulting from both growth and thermal stresses incorporated during cooling. PMID- 15348252 TI - Tissue responses to anti-washout apatite cement using chitosan when implanted in the rat tibia. AB - The tissue response to anti-washout apatite cement using chitosan (aw-AC(chi)) was evaluated by implanting aw-AC(chi) into bone defects of rat tibiae using conventional apatite cement (c-AC) as a control material. During the experimental period up to 16 weeks, the only difference between aw-AC and c-AC was found at two weeks in the tissue response of soft tissue. At two weeks, c-AC showed a moderate inflammatory response; small particles of c-AC were scattered in the cutaneous tissue and many foreign body giant cells were collected around the scattered c-AC, whereas aw-AC showed only a slight inflammatory response and few foreign body giant cells. We found no difference between aw-AC(chi) and c-AC with respect to bone tissue response. Both AC were almost completely surrounded by mature bone at eight weeks. No promotion or reduction of osteoconductivity was observed by chitosan even though it is considered to promote bone formation. We concluded, therefore, that enhancement of bone formation cannot be expected by employing chitosan to obtain anti-washout properties, at least in the concentration used in this study, even though aw-AC(chi) is much more useful than c-AC. PMID- 15348253 TI - Tissue response to hafnium. AB - The aim of the present experimental study was to evaluate the tissue response to hafnium (Hf) a reactive metal closely related to titanium (Ti) and zirconium (Zr). Hf has not been previously evaluated as implant material in a biologic environment. In a first experiment, 21 machined Hf non-threaded implants (test) and 21 similar Ti implants (control) were inserted in the abdominal wall of 21 rats. Animals were sacrificed after 8 days (6 rats), 6 (7 rats) and 12 weeks (8 rats). In a second experiment, 18 rabbits received 18 Hf and 18 Ti threaded implants in their tibiae, one implant in each tibia. The rabbits were sacrificed after 6, 12 and 24 weeks (6 animals/time interval). The bulk metal of the abdominal wall implants, embedded together with the surrounding tissue, was electrolytically dissolved and semithin (1 microm) sections of the intact tissue implant interface were evaluated by light microscopy (morphometry). Bone-implant contact and bone area within threads were evaluated in ground sections. In soft tissues, a fluid space containing predominantly monocytes/macrophages surrounded the abdominal implants at 8 days. At 6 and 12 weeks, a fibrous capsule, consisting of layers of macrophages and fibroblasts, surrounded the implants. Macrophages, including multinuclear giant cells, always formed the innermost layer in contact with the implant surface. No quantitative or qualitative difference in the tissue organization was detected between Ti and Hf implants. In rabbits, 6 weeks after insertion, the proximal two threads located within the cortical bone were filled with bone in contact with Hf and Ti. The distal threads contained bone marrow. After 12 and 24 weeks, mature bone was present in the proximal 3-4 implant threads. No statistically significant difference was found between Hf and Ti implants at any time periods. It is concluded that Hf is an interesting metal for biomedical applications in bone and soft tissue. PMID- 15348254 TI - Endothelial cell adhesion on bioerodable polymers. AB - This paper presents the results of a preliminary screening of a new class of bioerodable polymers, partial esters of alternating copolymers of maleic anhydride and mono-methoxyoligoethyleneglycol vinyl ethers (PAM) for use in engineered vascular tissue. Different initial concentrations of PAM and human serum albumin (HSA) were spin-coated onto glass substrates and the surface properties of the resulting films and their relationship to endothelial cell adhesion was examined. An optimum PAM/HSA blend for use as the cell contact surface of a bioerodable scaffold was identified. PMID- 15348255 TI - The effect of molecular orientation and acetylene-enhanced crosslinking on the wear of UHMWPE in total artificial joints. AB - This paper investigates the benefits of combining roll-drawing and acetylene enhanced crosslinking to alter the mechanical properties of the ultra high molecular weight polyethylene (UHMWPE) used in total hip and knee replacements, with the aim of improving its resistance to wear. UHMWPE was processed via crosslinking, roll-drawing and a combination of crosslinking and roll-drawing and subjected to gel content analysis, tensile tests, X-ray diffraction and wear tests using different types of motion and smooth and rough counterfaces. Purely roll-drawn materials with length and width draw ratios of lambda l x lambda w = 1.3 x 1.0 and lambda1 x lambdaw = 1.6 x 0.9 respectively, were found to have lower wear factors in a unidirectional motion test with a rough counterface when compared to the virgin material. The crosslinked roll-drawn material, with length and width draw ratios of lambda1 x lambdaw = 1.6 x 0.9, was seen to possess five crosslinks per initial number average molecule. This crosslinked and roll-drawn material showed 5.5 times less wear than the virgin material in a multidirectional motion test with a smooth counterface and 1.4 times more wear than the virgin material in a unidirectional motion test with a rough counterface. Hence this study supports previous work by the authors that acetylene-enhanced crosslinked materials may show benefits for a total hip replacement, but only where the femoral head remains smooth. The improvements in wear with the roll-drawn material in unidirectional tests were smaller, but may prove to have some benefits in the knee. PMID- 15348256 TI - HA and double-layer HA-P2O5/CaO glass coatings: influence of chemical composition on human bone marrow cells osteoblastic behavior. AB - Human osteoblastic bone marrow derived cells were cultured for 28 days onto the surface of a glass reinforced hydroxyapatite (HA) composite and a commercial type HA plasma sprayed coatings, both in the "as-received" condition and after an immersion treatment with culture medium during 21 days. Cell proliferation and differentiation were analyzed as a function of the chemical composition of the coatings and the immersion treatment. Cell attachment, growth and differentiation of osteoblastic bone marrow cells seeded onto "as-received" plasma sprayed coatings were strongly affected by the time-dependent variation of the surface structure occurring during the first hours of culture. Initial interactions leading to higher amounts of adsorbed protein and zeta potential shifts towards negative charges appeared to result in surface structures with better biological performance. Cultures grown onto the pretreated coatings showed higher rate of cell proliferation and increased functional activity, as compared to those grown onto the corresponding "as-received" materials. However, the cell behavior was similar in the glass composite and HA coatings. The results showed that the glass composites present better characteristics for bone cell growth and function than HA. In addition, this work also provide evidence that the biological performance of the glass composites can be modulated and improved by manipulations in the chemical composition, namely in the content of glass added to HA. PMID- 15348257 TI - Biological properties of copolymer of 2-hydroxyethyl methacrylate with sulfopropyl methacrylate. AB - Interaction of organism with non-toxic implanted polymers depends on the physicochemical properties of the implant surface, which influence the adsorption of bioactive proteins and subsequently adhesion and growth of cells. The synthetic hydrogels are known as poorly adhesive surfaces. In this study we demonstrated the adsorption of albumin, fibrinogen, fibronectin, basic fibroblast growth factor, heparin-binding epidermal growth factor-like growth factor and epidermal growth factor to poly(2-hydroxyethyl methacrylate) (pHEMA) and copolymer of 2-hydroxyethyl methacrylate (HEMA) and potassium salt of 3 sulfopropyl methacrylate (SPMAK). The adhesion and growth of 3T3 cells and human keratinocytes on surface of these polymers was tested without and with pretreatment of polymers with heparin-binding epidermal growth factor-like growth factor. The adhesion of mixture of human granulocytes and monocytes to these surfaces was also tested. The strips of both polymers were subcutaneously and intracerebrally implanted into the rat and the extent of foreign body reaction and brain biocompatibility was evaluated. The results showed the extensive adsorption of basic fibroblast growth factor and heparin-binding epidermal growth factor-like growth factor to copolymer containing SPMAK. However the adhesion (and growth) of cells to this type of copolymers was very low. Preadsorption of human plasma to pHEMA clearly stimulated the leukocyte adhesion in contrary to copolymer containing SPMAK. The extent of foreign-body reaction was significantly higher against the pHEMA compared to tested copolymer p(HEMA-co-SPMAK). In conclusion, the tested copolymer was a poorly adhesive substrate that is only poorly recognized by the non-specific immunity, although the adsorption of basic growth factors to this substrate is highly significant. Both polymers were well tolerated by the brain tissue. The phenotype of surrounding neurons was more close to the control neurons in the brain tissue surrounding the p(HEMA-co-SPMAK) implants. PMID- 15348258 TI - Variations in the compressive strength of dental cements stored in ionic or acidic solutions. AB - The compressive strengths of various dental cements (a zinc polycarboxylate, a zinc phosphate, a glass-ionomer and two resin-modified glass ionomers, RMGICs) have been determined following storage in pure water, 0.9% sodium chloride solution or 20 mmol dm(-3) lactic acid solution for periods of time ranging from 24 h to 3 months. The glass-ionomer cement showed no differences between different storage solutions or at different storage times, whereas the zinc polycarboxylate, zinc phosphate and the resin-modified glass ionomer cements showed significant differences following storage in the solutions for 24 h compared with pure water. The zinc polycarboxylate cement was significantly weaker at 24 h in 0.9% NaCl and lactic acid than in pure water, whereas most of the other cements were significantly stronger in both 0.9% NaCl and lactic acid. One of the RMGICs (Vitremer luting, ex. 3M), however, was significantly stronger only in the NaCl solution, not in the lactic acid. In general, by 1 week, the strengths all reverted to being essentially the same as for specimens stored in pure water for most subsequent storage times, and did not change significantly on storage for up to 3 months. This effect of storage medium on the early strength has not been reported previously and since the media were chosen to model certain characteristics of natural saliva, the changes observed seem likely to occur in vivo. It is concluded that pure water is not the best medium for storing these cements if they are to behave as they do under clinical conditions. PMID- 15348259 TI - Apoptosis in interface membranes of aseptically loose total hip arthroplasty. AB - The terminal events leading to periprosthetic osteolysis are multifactorial in nature and modulation of this process after the stage of osteolytic mediator release has been futile. Recently, the demonstration of the ability of bisphosphonates to inhibit bone resorption that is mediated by particle stimulated macrophages and their induction of osteoclast apoptosis suggests a potent area for modulation of osteolysis at the prosthesis-bone interface. The purpose of this study was to determine the mode of cell death that occurs at the osteolytic interface of failed total hip arthroplasty (THA). TUNEL staining, DNA laddering, and immunodetection of poly(ADP-ribose)polymerase (PARP) protein were used to identify the presence of apoptosis in interface membranes from 25 patients aged 28-88 years old (mean, 58 years) harvested at the time of hip revision surgery. Our results demonstrated positive TUNEL stain in 100% of specimens with an average 37% of cells (range 12-60%) positively stained for TUNEL whereas less than 8% of control tissue cells showed positive staining. DNA laddering, a characteristic feature of apoptotic cells, was observed in 82% (28/34) of specimens studied at both the acetabular and femoral side of aseptically loose THAs. No laddering was observed in control tissues. Finally, using Western blot analysis, we observed the appearance of the 89 kDa PARP fragment associated with apoptosis in 92% of specimens (30/33). Our results demonstrate the presence of apoptotic cell death in interface membranes of THAs suggesting that apoptosis-related events are indeed associated with periprosthetic osteolysis and could serve as a specific target point for therapeutic modulation. PMID- 15348260 TI - Hydroxyapatite implants with designed internal architecture. AB - Porous hydroxyapatite (HA) has been used as a bone graft material in the clinics for decades. Traditionally, the pores in these HAs are either obtained from the coralline exoskeletal patterns or from the embedded organic particles in the starting HA powder. Both processes offer very limited control on the pore structure. A new method for manufacturing porous HA with designed pore channels has been developed. This method is essentially a lost-mold technique with negative molds made with Stereolithography and a highly loaded curable HA suspension as the ceramic carrier. Implants with designed channels and connection patterns were first generated from a Computer-Aided-Design (CAD) software and Computer Tomography (CT) data. The negative images of the designs were used to build the molds on a stereolithography apparatus with epoxy resins. A 40 vol% HA suspension in propoxylated neopentyl glycol diacrylate (PNPGDA) and iso-bornyl acrylate (IBA) was formulated. HA suspension was cast into the epoxy molds and cured into solid at 85 degrees C. The molds and acrylate binders were removed by pyrolysis, followed by HA green body sintering. With this method, implants with six different channel designs were built successfully and the designed channels were reproduced in the sintered HA implants. The channels created in the sintered HA implants were between 366 microm and 968 microm in diameter with standard deviations of 50 microm or less. The porosity created by the channels were between 26% and 52%. The results show that HA implants with designed connection pattern and well controlled channel size can be built with the technique developed in this study. PMID- 15348261 TI - A simple chemical method for preparation of hydroxyapatite coatings on Ti6Al4V substrate. AB - A novel, simple and economical chemical bath method for deposition of calcium hydroxyapatite coating has been developed. The coatings were prepared from EDTA solutions in alkaline media on Ti6Al4V substrates. The method is based on thermal dissociation of the Ca(EDTA)2- complex at 65-95 degrees C. Two chemical baths with and without presence of Na+ and Cl- were used for the deposition. The Rutherford back scattering study shows that the coating material from bath which contained sodium and chlorine ions have their presence in the coated material. The bath which has been prepared with potassium substituting sodium and nitrate substituting chlorine produced coatings with better stoichiometry, with Ca/P=1,67. The X-ray analysis revealed that the calcium hydroxyapatite coatings have preferred crystal orientation in the 002 direction. PMID- 15348262 TI - Numerical simulation of the casting process of titanium tooth crowns and bridges. AB - The objectives of this paper were to simulate the casting process of titanium tooth crowns and bridges; to predict and control porosity defect. A casting simulation software, MAGMASOFT, was used. The geometry of the crowns with fine details of the occlusal surface were digitized by means of laser measuring technique, then converted and read in the simulation software. Both mold filling and solidification were simulated, the shrinkage porosity was predicted by a "feeding criterion", and the gas pore sensitivity was studied based on the mold filling and solidification simulations. Two types of dental prostheses (a single crown casting and a three-unit-bridge) with various sprue designs were numerically "poured", and only one optimal design for each prosthesis was recommended for real casting trial. With the numerically optimized design, real titanium dental prostheses (five replicas for each) were made on a centrifugal casting machine. All the castings endured radiographic examination, and no porosity was detected in the cast prostheses. It indicates that the numerical simulation is an efficient tool for dental casting design and porosity control. PMID- 15348263 TI - Osteoblast responses to as-deposited and heat treated sputtered CaP surfaces. AB - The clinical success of dental implants is governed by implant surfaces and bone cell responses that promote rapid osseointegration. The objective of this study was to evaluate the in vitro osteoblast cell response to heat treated and non heat treated CaP coatings. In this study, the heat treated surfaces exhibited a poorly crystallized HA-type structure whereas the non-heat treated surface exhibited an amorphous structure. The heat treated CaP surfaces were observed to have a mean contact angle measurement of 57.95+/-0.95 degrees, whereas the non heat treated CaP surfaces were observed to have a mean contact angle measurement of 44.6+/-0.3 degrees. From the in vitro cell culture study, the ATTC CRL 1486 human embryonic palatal mesenchyme (HEPM) cells displayed a similar protein production and hexosaminidase activity on the heat treated and non-heat treated CaP surfaces throughout the nine day experiment. However, the HEPM cells cultured on non-heat treated CaP surfaces were observed to have higher specific ALP activity after nine days' incubation compared to cells cultured on heat treated CaP surfaces. The higher specific ALP activity by cells on non-heat treated surfaces were suggested to be attributed to the lower degree of crystallinity and the lower contact angles observed in this study. PMID- 15348264 TI - Nano-crystallization and surface analysis of electrolytic ZrO2 coatings on Co-Cr alloy. AB - Zirconia coatings were formed on Co-Cr substrates by electrolytic deposition. The microstructure of electrolytic zirconia-coated films on Co-Cr substrates was examined. According to the results of ESCA, the bonding energies of ZrO2 coating surface layer which changed with the annealing temperature from 400 degrees C, 500 degrees C to 600 degrees C are attributed to amorphous (a), tetragonal (t) and monoclinic (m) structure, respectively. The X-ray diffraction (XRD) of the coatings on the Co-Cr substrates annealed at 400, 500 and 600 degrees C revealed the major crystallization from m through t and then to m+t. However, TEM observations clearly showed that the interface layer of the coatings were nanosize crystallites, first the formation of tetragonal and monoclinic ZrO2 structures. These different phase transformations are mainly due to the different surface energy of ZrO2 coating in air, in bulk or on Co-Cr alloy. PMID- 15348265 TI - Release of fluoride from glass fiber-reinforced composite with multiphase polymer matrix. AB - This study investigated the suitability of a fluoride containing monomer resin system for use as a copolymer of dental fiber-reinforced composite (FRC) materials. The purpose of the study was to measure the release of fluoride from the test materials. The monomer resin system was either light-polymerized, or light-polymerized and post-cured with heat at 130 degrees C. The release of fluoride from FRC test specimens during 30 day storage periods was compared to the release of fluoride from unreinforced test specimens (n = 5). The fluoride release into distilled water was determined with an ionanalyzator. During the first week of water storage, the fluoride release was 0.31 wt % for the unreinforced specimens and 0.13 wt % for reinforced specimens. The post-curing had no influence on the fluoride release values. The results of this study suggest that fiber inclusion reduces fluoride release of reinforced specimens compared to unreinforced specimens because the amount of polymer was smaller in reinforced specimens. The results of this study showed that the fluoride containing monomer system could be incorporated into the polymer matrix of fiber reinforced composites. PMID- 15348266 TI - Local mechanical and optical properties of normal and transparent root dentin. AB - The mechanical and optical properties of healthy and transparent root dentin are compared using atomic force microscopy (AFM), micro-Raman and emission spectroscopies and fluorescence microscopy. The elastic modulus and hardness of intertubular and peritubular transparent and healthy dentin did not differ appreciably. The tubule filling material in the transparent zone, however, exhibited values between peritubular and intertubular dentin. Raman spectroscopy revealed a shift in the 1066 cm(-1) band to 1072 cm(-1) from normal to transparent intertubular dentin. The material filling the tubule lumen in transparent dentin showed an increase in frequency of the band near 1070 cm(-1) as well. The emission spectral characteristics under 351 nm photoexcitation indicate differences between normal and transparent intertubular dentin. A transition region of about 300 microm between normal and transparent dentin was identified. In this region the intertubular emission properties were the same as for normal dentin, but tubules were filled. The filling material had emission characteristics closer to the normal intertubular than to transparent intertubular dentin. PMID- 15348267 TI - Effect of heat treatment on bioactivity and mechanical properties of PDMS modified CaO-SiO2-TiO2 hybrids via sol-gel process. AB - Crack- and pore-free transparent monolithic disks of polydimethylsiloxane (PDMS)- modified CaO-SiO2-TiO2 hybrids were obtained by hydrolysis and polycondensation of PDMS, tetraethoxysilane, tetraisopropyltitanate and calcium nitrate. The product as-dried at 60 degrees C formed an apatite on its surface in a simulated body fluid (SBF) within only one day, indicating its high bioactivity. The apatite-forming ability decreased slightly by a heat treatment below 250 degrees C. The bending strength of the product was about 11 MPa, independent of the heat treatment. This average strength value is comparable to that of the human cancellous bone. Young's modulus of the products increased from 100 to 500 MPa with increasing heat treatment temperature from 60 to 250 degrees C, but its values were within the range of those of the human cancellous bone. The strain at failure of the products decreased with increasing heat treatment temperature. Failure strains went down to the magnitudes exhibited by the human cancellous bone, when the products were heat treated in the temperature range from 150 to 250 degrees C. Thus, highly bioactive hybrids with mechanical properties analogous to those of the human cancellous bone were obtained. This new kind of bioactive hybrid may be useful as a bone-repairing material. PMID- 15348268 TI - Microstructuring ceramic scaffolds for hepatocyte cell culture. AB - Both extracorporeal liver support devices and tissue engineering of liver for transplantation require the maintenance of functionality of liver cells (hepatocytes) in cell culture for a long time. One approach to achieve this is to optimize hepatocyte in vitro environment by using a scaffold with topographic structure at sub-millimeter scale which controls cell distribution. Therefore, a set of new type of titania ceramic scaffolds, containing cavities of several sizes, has been produced for deducing the best choice of cavity dimensions for culturing hepatocytes. The aim of this paper is to describe in detail the production methods and characterization of such ceramic scaffolds. Experimental production of the scaffolds consists of microfabrication of silicon templates as well as preparation and molding of titania ceramics. The templates, containing arrays of conical protrusions arranged in close-packed hexagonal order, have been achieved using microfabrication methods of photolithography and anisotropic etching in KOH at 50 degrees C. Protrusion dimensions and overall quality of the templates has been evaluated by scanning electron microscopy. The microfabricated templates have resulted in well-defined and reproducible cavities of corresponding dimensions on the titania ceramic surface after injection-molding. Alternatively, simple embossing of the plastified green ceramics with the silicon templates attached to a metal plate also creates cavities on the ceramic surface. While both methods yield good results, they have different advantages: the injection-molding provides a higher quality of imprints while embossing is quicker and less complicated, and is not limited by dimensions of specific molding equipment. PMID- 15348269 TI - Biomimetic coatings on titanium: a crystal growth study of octacalcium phosphate. AB - The biomimetic approach allows the coating of metal implants with different calcium-phosphate (Ca-P) phases. Films elaborated at physiological conditions exhibited structures closely resembling those of bone mineral. For instance, octacalcium phosphate (OCP, Ca8(HPO4)2(PO4)4 . 5H2O) crystals have been deposited on titanium through a two-step procedure. After cleaning and etching, Ti6Al4V plates were immersed for 24 h into a simulated body fluid (SBF1). A thin amorphous carbonated Ca-P layer precipitated on the metal substrate. Secondly, these thinly Ca-P coated titanium substrates were immersed for 48 h into another simulated body fluid (SBF2). The thin amorphous carbonated Ca-P layer induced the fast precipitation of a second Ca-P layer of 55 microm in thickness composed of OCP crystals. The measurements of Ca and P concentrations versus soaking time in SBF2 showed that the carbonated Ca-P layer partially dissolved before the deposition of the OCP coating. X-ray diffraction (XRD) revealed that OCP crystals grew epitaxially on the substrate. OCP is known to be one of the precursors during the bone mineralization process, thereby, this new generation of biomimetic coatings are promising for orthopedic surgery. PMID- 15348270 TI - Incorporation of salicylic acid derivatives to hydrophilic copolymer systems with biomedical applications. AB - Hydrogels based on polymeric derivatives of salicylic acid have been prepared for biomedical applications by free radical copolymerization of 2-hydroxy-4 methacrylamidobenzoic acid, 4HMA, and 2-hydroxy-5-methacrylamidobenzoic acid, 5HMA, with 2- hydroxyethylmethacrylate, HEMA, in a wide range of compositions. The reactivity ratios of 4HMA and 5HMA with HEMA in radical copolymerization processes have been determined from their 1H NMR spectra by applying linearization methods and non-linear least square treatments. Tgs of the corresponding copolymers were analyzed by DSC. The swelling behavior in water of the prepared copolymers was studied in comparison to poly-(HEMA), poly-(4HMA) and poly-(5HMA) hydration degrees, being in all cases superior to 35%. The hydrolytical behavior of the synthesized copolymers was studied at three different pHs (2, 7.4 and 10) determining the release percentage of the salicylic acid derivatives, 4-amino salicylic acid, 4ASA, and 5-amino salicylic acid, 5ASA, analyzed by high performance liquid chromatography (HPLC). The release analysis was followed during 230 days and a pH dependence was observed obtaining the highest release percentages at pH=10, whereas at physiological pH (7.4) the release percentages were in range from 2 to 5% at that time for all copolymer systems. The hydrolytical stability is enough for long-term applications like bone cements, ionomers, etc. PMID- 15348271 TI - Staphylococcus epidermidis RP62A adhesion to chemically modified cellulose derivatives. AB - The adhesion of coagulase negative Staphylococcus epidermidis expressing capsular polysaccharide/adhesin (PS/A) to cellulose diacetate (CDA), as well as to primary reference low-density polyethylene, was assessed in vitro. Attached bacteria were released by gentle sonication and quantified as colony forming units. Surface free energy of cells and materials and the free energy of interaction between cells, each type of material and water molecules was calculated through contact angle measurement, also enabling the determination of materials surface hydrophobicity. The influence of CDA surface modification by deacetylation and phosphorylation on bacterial adhesion was studied. Chemical modifications of CDA by deacetylation and by phosphorylation were effective in lowering bacterial adhesion. PMID- 15348272 TI - Chemical and biological characteristics of low-temperature plasma treated ultra high molecular weight polyethylene for biomedical applications. AB - Several low-temperature radio-frequency (RF) plasma surface treatments were performed on ultra-high molecular weight polyethylene (UHMWPE) used in biomedical applications. Process gases included Ar, C3F6, CH4, hexamethyldisiloxane (HMDSO), and NH4. These treatments were carried out at pressures in the range of 64-400 mTorr, RF powers of 240-1200 W, and temperatures well below the melting point of UHMWPE. X-ray photoelectron spectroscopy (XPS) was used to obtain information about the surface characteristics of UHMWPE treated with the HMDSO, C3F6, and CH4 gases as a function of treatment conditions. XPS spectra of UHMWPE treated with C3F6 and CH4 and exposed to a laboratory environment for different time periods were examined in order to assess the stability of these treatments. It was found that for the C3F6 process gas the amount of fluorine at the surface decreased over time, whereas the oxygen content of the CH4 treated samples increased as a function of time. In vitro cytotoxicity of Ar, C3F6, CH4, and NH4 plasma treated samples was studied in light of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test results. The hemolytic nature of the various plasma treatments was evaluated using standard hemolysis tests. All of the samples tested in this study exhibited no cytotoxic and negligible hemolytic effects. The process parameters for several low-temperature plasma treatments demonstrating chemical and structural stability and good biocompatibility are discussed in conjunction with the broad applicability to other biomedical polymers. PMID- 15348273 TI - Effect of HIPping, stress and surface finish on the environmental degradation of Y-TZP ceramics. AB - Hot isostatically pressed (HIPped) and hot pressed (designated unHIPped) bar samples of yttria stabilized tetragonal zirconia polycrystalline ceramic (3Y-TZP) were subjected to three point bend testing in water at 90 degrees C. HIPped femoral heads with three different surface finishes were also aged in 90 degrees C water. The early stages of the environment induced tetragonal to monoclinic transformation was monitored as a function of time by X-ray diffraction, white light interferometry and scanning electron microscopy. HIPped samples were found to transform less readily than unHIPped samples and have a longer incubation period prior to transformation. There was an increase in the amount of monoclinic phase detected following the application of stress, particularly on the compression surface. Microcraters, believed to result from the expulsion of transformed grains when stress was applied, were observed on tension surfaces, particularly of unHIPped samples. There was no effect of surface roughness on the environment induced transformation for the range of surface finishes investigated. PMID- 15348274 TI - Studies on cytotoxic effect of nickel ions on three cultured fibroblasts. AB - Cytotoxicity of Ni ions on three fibroblasts such as L929, Balb/3T3 clone A31 and MC3T3-E1 were examined by cell count (CC) and Neutral Red assay (NR). Three cells were incubated for 6 days in 1 ml DME medium containing Ni ions which ranged from 0 to 2 mM/l. The results clarified that Ni ions had dose-dependent cytotoxicity. L929 possessed the largest TC50 values (the amount of Ni ion that caused 50% cell death or 50% cell viability) of 0.12 mM/l (CC) and 0.32 mM/l (NR), and Balb/3T3 clone A31 had the least values of 0.05 mM/l (CC) and 0.09 mM/l (NR), whilst MC3T3 E1 had the intermediate values of 0.08 mM/l (CC) and 0.15 mM/l (NR). The dissolution of Ni ions from Ni-containing metallic restorations must be lower than these concentration levels so that body tissues might not be severely damaged. PMID- 15348275 TI - Surface modification of natural rubber latex films via grafting of poly(ethylene glycol) for reduction in protein adsorption and platelet adhesion. AB - Natural rubber (NR) latex films with surface grafted poly(ethylene glycol) (PEG) chains were prepared by UV-induced graft copolymerization of methoxy poly(ethylene glycol) monomethacrylate (PEGMA) onto the plasma-pretreated NR latex films. PEGMA macromononers of different molecular weights were used. The UV induced graft copolymerization of PEGMA onto the plasma-pretreated NR latex films was also explored with PEGMA of different macromonomer concentrations and with different UV graft copolymerization time. The surface microstructures and compositions of the PEG-modified NR latex films were characterized by contact angle, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM) measurements. In general, higher macromonomer concentration and longer UV graft copolymerization time led to a higher graft yield. Water contact angle measurements revealed that the hydrophilicity of the NR latex film surface was greatly enhanced by the grafting of the PEG chains. The NR surface with a high density of grafted PEG was very effective in reducing protein adsorption and platelet adhesion. A lower graft concentration of the high-molecular-weight PEG was more effective than a high graft concentration of the low-molecular-weight PEG in reducing protein adsorption and platelet adhesion. PMID- 15348276 TI - Macroporous biphasic calcium phosphate ceramics versus injectable bone substitute: a comparative study 3 and 8 weeks after implantation in rabbit bone. AB - Macroporous biphasic calcium phosphate ceramics (MBCP) and a calcium phosphate injectable bone substitute (IBS), obtained by the association of biphasic calcium phosphate (BCP) ceramic granules and an aqueous solution of a cellulosic polymer, were compared in the same animal model. The two tested biomaterials were implanted in distal femoral osseous defects in rabbits. Qualitative and quantitative histological evaluation was performed three and eight weeks after implantation to investigate bone colonization and ceramic biodegradation associated with the two bone substitutes. Both biomaterials expressed osteoconduction properties and supported the apposition of a well-mineralized lamellar newly-formed bone. Bone colonization occurred much earlier and faster for IBS than for MBCP implants, although the respective rates of newly-formed bone after eight weeks of implantation did not differ significantly. For both biomaterials, ceramic resorption occurred regularly throughout the implantation period, though to a greater extent with IBS than with MBCP implants. The associated polymer in IBS produced intergranular spaces allowing body fluids to reach each BCP ceramic granule immediately after implantation, which may have favored osteoblastic activity, new bone formation and ceramic resorption. This completely interconnected open macroporosity could account for the earlier and more satisfactory bone substitution achieved with IBS. PMID- 15348277 TI - Characterization of copper corrosion products originated in simulated uterine fluids and on packaged intrauterine devices. AB - This paper studies the characterization of corrosion products originated after 1 and 12 weeks' immersion of copper specimens in simulated uterine fluids at pH 6.3 and 8.0 and at 37 degrees C temperature. The experimental techniques used were X ray photo-electron spectroscopy, scanning electron microscopy, and energy dispersive X-ray. The compounds found were calcite (CaCO3), calcium phosphate, cuprite (Cu2O) and copper hydroxide (Cu(OH)2). The morphology of corrosion products was a non-uniform, layer showing some paths through which copper ions can be released. In parallel, corrosion products formed on packaged, unused copper-containing intrauterine devices (IUD) were analyzed. Cuprite (Cu2O) and chalcocite (Cu2S) were the main species identified. PMID- 15348278 TI - Conditioning film and environmental effects on the adherence of Candida spp. to silicone and poly(vinylchloride) biomaterials. AB - The reported incidence of colonization of oropharyngeal medical devices with Candida spp. has increased in recent years, although few studies that have systematically examined the adherence of yeast cells to such biomaterials, the primary step in the process of colonization. This study, therefore, examined the effects of oropharyngeal atmospheric conditions (5% v/v carbon dioxide) and the presence of a salivary conditioning film on both the surface properties and adherence of Candida albicans, Candida krusei and Candida tropicalis to PVC and silicone. Furthermore, the effects of the salivary conditioning film on the surface properties of these biomaterials are reported. Growth of the three Candida spp. in an atmosphere containing 5% v/v CO2 significantly increased their cell surface hydrophobicity and reduced the zeta potential of C. albicans and C. krusei yet increased the zeta potential of C. tropicalis (p<0.05). Furthermore, growth in 5% v/v CO2 decreased the adherence of C. tropicalis and C. albicans to both PVC and silicone, however, increased adherence of C. krusei (p<0.05). Pre treatment of the microorganisms with pooled human saliva significantly decreased their cell surface hydrophobicity and increased their adherence to either biomaterial in comparison to yeast cells that had been pre-treated with PBS (p<0.05). Saliva treatment of the microorganisms had no consistent effect on microbial zeta potential. Interestingly, adherence of the three, saliva-treated Candida spp. to saliva-treated silicone and PVC was significantly lower than whenever the microorganisms and biomaterials had been treated with PBS (p<0.05). Treatment of silicone and PVC with saliva significantly altered the surface properties, notably reducing both the advancing and receding contact angles and, additionally, the microrugosity. These effects may contribute to the decreased adherence of saliva-treated microorganisms to these biomaterials. In conclusion, this study has demonstrated the effects of physiological conditions within the oral cavity on the adherence of selected Candida spp. to biomaterials employed as oropharyngeal medical devices. In particular, this study has ominously shown that these materials act as substrates for yeast colonization, highlighting the need for advancements in biomaterial design. Furthermore, it is important that physiological conditions should be employed whenever biocompatibility of oropharyngeal biomaterials is under investigation. PMID- 15348279 TI - Preparation and characterization of calcium phosphate-poly(vinyl phosphonic acid) composites. AB - Composites of calcium phosphates and the calcium salt of a biomedical polymer were prepared in situ by hot-pressing particulate mixtures of poly (vinyl phosphonic acid) (PVPA) and tetracalcium phosphate Ca4(PO4)2O, or TetCP) at different temperatures, pressures, and time periods. The objective was to establish whether PVPA could react with TetCP (Ca/P ratio of 2.0) to form a calcium salt, and thereby decrease the available Ca/P ratio 1.67 to facilitate hydroxyapatite (Ca10(PO4)6(OH)2 or HAp) formation. The effects of varying the bulk composition, temperature (to 300 degrees C), pressure (to 690 MPa) and time (to 60 min) on the reaction between TetCP and PVPA were studied using X-ray diffraction, infra-red spectroscopy and scanning electron microscopy techniques. Results showed that the conversion of TetCP into HAp increased with compaction time as temperature and/or pressure were increased. Formation of anhydrous dicalcium phosphate (CaHPO4, or DCPA) was also observed. Complete conversion of TetCP to HAp was achieved in composites pressed at 250 degrees C and 415 MPa for 30 min. PMID- 15348280 TI - Structural characterization of the metal/glass interface in bioactive glass coatings on Ti-6Al-4V. AB - Coating Ti-based implants with bioactive materials promotes joining between the prostheses and the bone as well as increasing long-term implant stability. In the present work, the interface between Ti-6Al-4V and bioactive silicate glass coatings, prepared using a simple enameling technique, is analyzed. High resolution transmission electron microscopy of the glass/alloy interface shows the formation of a reaction layer ( approximately 150 nm thick) composed of Ti5Si3 nanoparticles with a size of approximately 20 nm. This nanostructured interface facilitates the formation of a stable joint between the glass coating and the alloy. PMID- 15348281 TI - Inflammatory response to a porcine membrane composed of fibrous collagen and elastin as dermal substitute. AB - The inflammatory response to a collagen/elastin membrane was studied by measuring the expression of cytokines and function associated antigens in human macrophages. Additionally the angiogenic and inflammatory activity in the chorioallantoic membrane of the chick embryo (CAM-assay) was investigated. Macrophages cultured on the membrane expressed IL-1beta mRNA as early as after 4 hours. During prolonged culturing IL-1beta mRNA levels decreased. Messenger RNA for IL-8 was detectable over the whole culture period. The anti-inflammatory cytokine IL-10 was expressed up to one day only. Phenotypic analysis revealed a decrease in the number of chronic inflammatory 25F9 positive macrophages not migrating into the membrane but a presence of these cells together with the acute inflammatory 27E10 macrophages within the membrane whereas the anti-inflammatory subtype RM3/1 was absent. In the CAM-assay the membrane stimulated angiogenesis and induced the formation of granulation tissue. Histological analysis showed that the membrane was infiltrated with macrophages, fibroblasts and endothelial cells and locally with granulocytes. These data show that the collagen/elastin membrane causes activation of macrophages, angiogenesis and the formation of inflammatory tissue. Although these processes are essential for wound healing the type of inflammation points to a chronic process which might counteract an efficient scar formation. PMID- 15348282 TI - Porcine pericardial membrane subjected to tensile testing: preliminary study of the process of selecting tissue for use in the construction of cardiac bioprostheses. AB - The durability of cardiac bioprostheses is limited fundamentally by structural failure due to mechanical fatigue and calcification. In the present report, we analyze, using an in vitro hydraulic simulator to test tensile strength, the mechanical behavior of porcine pericardium for the purpose of establishing the criteria for selecting the biomaterial, taking into account both morphological criteria (thickness and homogeneity of the specimens) and mechanical criteria (stress at breaking point), using the epidemiological model of paired samples. The stress at breakage was found to range widely from 24.07 MPa to 100.29 MPa, although we observed no statistically significant differences when comparing the mean results in the different regions and zones of the pericardium being studies. The application of the selection criteria in the present series resulted in an excellent mathematical fit in terms of the stress/elongation (R2 > 0.95), making it possible to establish, by means of linear regression, the prediction of the tensile strength in one zone on the basis of the values observed in its paired specimen. PMID- 15348283 TI - In vivo evaluation of bone tissue behavior on ion implanted surfaces. AB - The ion implantation process offers several unique advantages over other surfaces modifications techniques, in regard to ion release and material mechanical characteristics. The aim of this study was to evaluate the in vivo bone tissue response to ion implanted surfaces. Untreated and nitrogen-ion-implanted stainless steel implants were inserted in the tibia diaphysis (cortical bone) and proximal tibia epiphysis (trabecular bone) of 12 New Zealand White rabbits. The animals were divided into three groups of four animals each, which were maintained for 4, 12 and 24 weeks according to internationally accepted and standardized procedures. At sacrifice, the implants were retrieved with surrounding bone and fixed in 4% neutral buffered formaldehyde and embedded in polymethylmethacrylate (PMMA). The samples were reduced in slices and stained with hematoxylin-eosin, light-green, fuchsin acid and giemsa solution for histological evaluation; fluorescent markers were also used to assess bone apposition. Histomorphometric evaluation was used to determine the extent of bone material contact. Results from histological and morphometrical analyses revealed active remodeling of bone around both types of implants (control and ion implanted). However, faster bone deposition was observed around the treated material (12 weeks). Both materials reached similar endpoints, as no significant differences between them were evident at 24 weeks. The results demonstrate that ion implanted stainless steel has similar, or slightly enhanced, biological compatibility in contact with bone compared to untreated material; thus it may be a useful material in biomedical applications where reduced ion release or enhanced mechanical properties (as provided by ion implantation) are required. PMID- 15348284 TI - Cross-linking by 1-ethyl-3- (3-dimethylaminopropyl)-carbodiimide (EDC) of a collagen/elastin membrane meant to be used as a dermal substitute: effects on physical, biochemical and biological features in vitro. AB - Next to in vitro-cultured autogeneic keratinocytes for the restoration of epidermis, a suitable dermal matrix is a mandatory component of an artificial skin substitute for the permanent covering of full thickness skin defects. In our model a xenogeneic membrane, consisting of processed native collagen and elastin of porcine origin is meant to serve as a template for the formation of a neo dermis. In order to improve the resistance of this matrix against enzymatical degradation, we cross-linked it by using the carbodiimide 1-ethyl-3-(3 dimethylaminopropyl)-carbodiimide (EDC) together with N-hydroxysuccinimide. Chemical cross-linking by these agents at two different degrees (shrinkage temperatures 63 degrees C and 81 degrees C) had no relevant effect on mechanical features or water-uptake capacity. The time needed for enzymatic digestion was increased by cross-linking. Concerning growth and spreading of fibroblasts and keratinocytes on and within the structure of this membrane, we did not observe a difference between cross-linked and non-cross-linked material (shrinkage temperature 48 degrees C). We therefore expect that cross-linking by EDC is an effective means to control the degradation of the collagen/elastin membranes in vivo without a significant influence on their biocompatibility. PMID- 15348285 TI - Surface modification of polyurethane for promotion of cell adhesion and growth 1: surface photo-grafting with N,N-dimethylaminoethyl methacrylate and cytocompatibility of the modified surface. AB - Functional polyurethane (PU) surface was prepared by photo-grafting N,N dimethylaminoethyl methacrylate (DMAEM) onto the membrane surface. Grafting copolymerization was conducted by the combined use of the photo-oxidation and irradiation grafting. PU membrane was photo-oxidized to introduce the hydroperoxide groups onto the surface, then the membrane previously immersed in monomer solution was irradiated by UV light. The X-ray photoelectron spectroscopy and water contact angle characterized the grafted copolymers and verified the occurrence of graft copolymerization. The results showed that UV irradiation could realize the graft copolymerization effectively. The grafted membrane showed minimal surface morphology. Human umbilical vein endothelium (HUVE) cells were seeded on the grafted surfaces. The performance of the surface in cell attachment correlated with the content of oxygen and nitrogen. Cells were spread more extensive and grown faster on the surface with lower degree of grafting. PMID- 15348286 TI - Comparison of the material properties of PMMA and glass-ionomer based cements for use in orthopaedic surgery. AB - The intrinsic benefits of low exotherm and bioactivity have generated interest in utilizing glass-ionomer cements (GIC) as a bone cement replacement in orthopaedic surgery. This paper is concerned with evaluating the mechanical properties of compressive strength, flexural strength, and fracture toughness for two traditional GICs, one resin-modified GIC (an experimental bone cement) and two polymethylmethacrylate (PMMA) cement systems. To determine the suitability of a GIC system for use in the clinical orthopaedic setting, the additional characteristics of setting exotherm and setting time have also been evaluated. The characterization of these two vastly different cement systems has raised some concern as to the applicability of using the current orthopaedic standards for the testing of GIC systems. In particular, issues relating to the strain rate dependence of PMMA cement and the exothermic basis for determining setting time are not applicable as these factors are not characteristic of GIC systems. Whilst the intrinsic benefits of current GIC systems are well understood and generally accepted, this study has shown their intrinsic mechanical properties to be inferior to current PMMA cements. Improvement in the mechanical properties of traditional GICs have been achieved with the addition of a resin component (HEMA). PMID- 15348287 TI - The influence of calcium to phosphate ratio on the nucleation and crystallization of apatite glass-ceramics. AB - The nucleation and crystallization behavior of a series of glasses based on 4.5SiO2-3Al2O3-YP2O5-3CaO-1.51CaF2 was studied. The parameter Y was varied to give calcium to phosphate ratios between one and two. All of the glasses studied crystallized firstly to fluorapatite (Ca5PO4)3F). The glass with a calcium to phosphate ratio of 1.67, corresponding to apatite, bulk nucleated to give fluorapatite (FAP). The glasses with calcium : phosphate ratios either less than that of apatite, or greater than that of apatite all exhibited surface nucleation of FAP. However, following a nucleation hold of one hour at approximately 50 K above the glass transition temperature these glasses exhibited bulk nucleation of FAP. PMID- 15348288 TI - Evaluation of mechanical and corrosion biocompatibility of TiTa alloys. AB - As-received and heat-treated Ti40Ta and Ti50Ta alloys were evaluated to determine their corrosion as well as mechanical performances and compared to Ti6A14V, a common material utilized for orthopedic (surgical) implants. Anodic potentiodynamic tests performed in Plasmalyte showed that all samples, except for the Ti50Ta specimen aged at 400 degrees C for 3 h gave a curve similar to that of Ti6A14V. Optical and TEM microscopy was performed to determine as-received and heat-treated microstructures. As-received materials showed an alpha precipitate in an alpha+beta and martensite matrix. Samples that were aged at 400 degrees C increased in the density and the length of the alpha precipitate. Vickers hardness measurements were performed to get an approximation of the tensile strengths. Aged Ti40Ta and Ti50Ta specimens produced the highest tensile values when compared to the Ti6A14V material, representing a 31% and 56% increase for the 3 h samples and an 18% and 58% increase for the 10 h samples. Of all the materials studied the Ti50Ta specimen aged for 10 h exhibited the best biocompatibility showing excellent corrosion resistance combined with the highest tensile strength (1089 MPa and 58% harder/stronger than Ti6A14V). PMID- 15348289 TI - Adsorption of vascular endothelial growth factor to two different apatitic materials and its release. AB - The aim of our study was to assess the ability of calcium phosphate powders to serve as growth factor carriers. Vascular endothelial growth factor (VEGF), in particular, is locally involved in the bone formation process throughout osteoblast differentiation. Two different apatitic substrates were tested: hydroxyapatite (HA), widely used as biomaterial, and nanocrystalline carbonated apatite (CA), which has a composition similar to bone mineral crystals. These materials have been compared for their VEGF adsorption and release properties. The adsorption of the growth factor was higher on CA than on HA probably due to differences of both the proteins and the powders involved. The specific activity of the VEGF released was also tested to determine the available activity for cells in contact with these materials. Interestingly, the bioactivity of the VEGF released from CA quantified on fetal bovine aortic endothelial cells (FBAE) by evaluating the proliferation activity, exhibited no marked difference compared to native VEGF. Qualitatively, VEGF adsorbed on CA material induced well-defined collagen type I immunostaining on osteoblast cells compared to the staining obtained after VEGF adsorption on HA. PMID- 15348290 TI - Pin-on-plate studies on the effect of rotation on the wear of metal-on-metal samples. AB - An important cause of osteolysis and subsequent loosening of replacement hip joints is the body's biological response to polyethylene wear debris. Interest has thus been renewed in hard bearing surfaces such as metal-on-metal implants. Tests were performed on a pin-on-plate machine to determine the effects of pin rotation on the wear of two different compositions of cobalt chrome molybdenum (CoCrMo) against itself (high carbon and low carbon). With reciprocating motion only, the low carbon material gave an order of magnitude higher wear than the high carbon material. The overall wear (that for both the pin and the plate) was significantly reduced with added rotation for the low carbon material but remained approximately the same for the high carbon material. However, the wear of the low carbon material was not reduced below that of the high carbon material which remained the best material in terms of wear. PMID- 15348291 TI - Hydroxyapatite spheres with controlled porosity for eye ball prosthesis: processing and characterization. AB - Porous hydroxyapatite spheres were prepared by a modified gelcasting method producing a ceramic prosthesis with controled porosity. The spheres are approximately 2.2 cm in diameter with a relatively homogeneous pore size distribution from 10 to 40 microm in diameter. The samples were characterized by X-ray powder diffractometry (XRD) and Fourier transform infra-red spectroscopy (FTIR) to identify the phases both prior to and after the gelcasting process. Surface morphology analysis and porosity evaluations were performed with scanning electron microscopy (SEM), while surface area measurements were carried out by the BET technique. PMID- 15348292 TI - Release of 5-fluorouracil by biodegradable poly(ester-ether-ester)s. Part I: release by fused thin sheets. AB - The 5-fluorouracil release by biodegradable epsilon-caprolactone and L-lactide copoly(ester-ether-ester)s was tested. The drug-copolymer mixture was formed by fusion in thin sheets, which were dipped in Dulbecco's PBS for time intervals ranging from one hour to two months. Each experiment shows a fast initial release, which subsequently slows down and stops at a limiting value, depending on the copolymer composition. This behavior was attributed to an extraction of the drug present on the sheet surface, due only to its shape, and to hydrogen bonds between the drug and the copolymers. The results obtained lead to a possibility of using such copolymers as "time-delayed" drug-releasing systems, when formed in specimens with smaller surface-to-volume ratio, which could minimize the fast initial extraction. PMID- 15348293 TI - Combination of 3D MRI and connectivity analysis in structural evaluation of cancellous bone in rat proximal femur. AB - Three dimensional (3-D) magnetic resonance (MR) microimaging combined with connectivity analysis was tested in the study of the structure of cancellous bone. MR microimaging was performed in vitro with an average resolution of 20*20*35 microm. A 3-D connectivity analysis was used to model the trabecular bone as a network consisting of nodes and struts. Size distribution curves of these two structural elements and the interconnectivity of nodes was used to estimate the cancellous structure. The analysis suggested the occurrence of two simultaneous network structures in cancellous bone differed by the size of details. The degradative effect of ostcopenia is found to be slightly different in these two subsystems. Interconnectivity is seen to increase with the size of a node and the expected loss of connectivity due to ostcopenia is observed. The method described offers a new way in the topological estimation of interconnected medium. PMID- 15348294 TI - Technique for estimating fracture resistance of cultured neocartilage. AB - Tensile tests on an artificial tissue of cultured neocartilage are described with a view to obtaining objective criteria for the onset of fracture and to estimating the tissue fracture resistance. The energy dissipation and compliance of the hyperelastic, hysteretic load-displacement responses exhibit great sensitivity to crack propagation during loading. A simple scheme is presented for deconvoluting the energy dissipated by cracking from an overall hysteresis loop and thus estimating the fracture resistance. The results suggest that the apparent fracture resistance of cultured neocartilage is determined by events localized to the crack and may exhibit a transient increase on initial crack extension. PMID- 15348295 TI - Coating of hydroxyapatite on various substrates via hydrothermal reactions of Ca(edta)2- and phosphate. AB - Hydroxyapatite was coated on various substrates such as 12 mol % ceria-doped tetragonal zirconia (12Ce-TZP), 3 mol % yttria-doped tetragonal zirconia (3Y TZP), alumina, monetite coated titanium (Ti/CaHPO4) and calcium titanate coated titanium (Ti/CaTiO3) via hydrothermal reactions of Ca(edta)2- and 0.05 M NaH2PO4 at initial pH 6 and 160-200 degrees C for 0.5-6 h. Rod-like particles of hydroxyapatite precipitated to form film on the substrates above 160 degrees C. The morphology of the film changed significantly depending on the characteristics of substrate, i.e. hydroxyapatite entirely coated the surfaces of 12Ce-TZP, Ti/CaHPO4 and Ti/CaTiO3 plates, but sparsely deposited on 3Y-TZP and Al2O3 plates. Film thickness increased with time (ca. 20 and 90 microm on 12Ce-TZP plates for 0.5 and 6 h, respectively, at pH 6 and 200 degrees C). The adhesive strength of the film for the substrate was in the order, 12Ce TZP/hydroxyapatite(28 MPa) > Ti/CaTiO3/hydroxyapatite (22 MPa) > Ti/CaHPO4/hydroxyapatite (9 MPa). PMID- 15348296 TI - Preparation of a bone-like apatite foam cement. AB - The preparation of a porous bone-like calcium deficient apatite implant material was investigated. A novel cement system composed of an equimolar mixture of Ca4(PO4)2O, Ca(HPO4)2 . H2O, and CaCO3 was used. At a liquid/powder ratio of 0.83 ml/g low density open framework foam cements were formed due to the rapid evolution of CO2. The initial product of the reactants was CaHPO4 . H2O which then reacted with Ca4(PO4)O, forming a calcium deficient carbonated apatite, upon soaking of the cement blocks in SBF. Foam-like cements were composed of a plate like apatite due to epitaxial overgrowth and conversion of the brushite plate precursor. Cylinders of the foam cement were reinforced with an outer layer of a solid apatite cement to form a material suitable for application as a bone section implant. PMID- 15348297 TI - Evaluation of endothelial cell integrins after in ? vitro contact with polyethylene terephthalate. AB - The aim of this research was to evaluate the effect of polyethylene terephthalate (Woven Dacron) on the expression of endothelial integrins. Human umbilical vein endothelial cells were cultured on the material for 24 h. The integrins VLA-2 (alpha2beta1-CD49b/CD29), receptor for laminin and collagen, VLA-5 (alpha5beta1 CD49e/CD29), receptor for fibronectin, VLA-6 (alpha6beta1-CD49f/CD29), receptor for laminin, and alphaVbeta3-CD51/CD61 (receptor for vitronectin) were evaluated by flow cytometry. After contact with polyethylene terephthalate, a slight but significant decrease in the percentage of both CD29 and CD49e positive cells was observed, which suggests a lower number of cells expressing the fibronectin receptor alpha5beta1. Moreover, a significant increase in the mean channel for CD49b and for the vitronectin receptor CD51/CD61 was observed. The reduction in the fibronectin receptor could account for the poor endothelialization observed in vivo on polyethylene terephthalate. The increased expression of the vitronectin receptor, favoring the migration of smooth muscle cells, could give some information about the pathogenesis of intimal hyperplasia, which is a complication of vascular grafts. PMID- 15348298 TI - Human monocyte-derived macrophages and dendritic cells as targets for biomaterial cytocompatibility studies using an improved in vitro culture system. AB - Since macrophage plays a key role in the biocompatibility process, neoplastic macrophage cell lines and human blood monocytes are commonly used as target cells for in vitro biomaterial tolerance evaluation. However, tumor cells profoundly differ from normal tissue cells and monocytes are only precursors of macrophages. It has become possible to generate recently, under adherent-free conditions, fully mature macrophages and dendritic cells from human blood monocytes in the presence of GM-CSF and GM-CSF + IL4 respectively. In the present work, we examined the effects of titanium-alloy on morphology, adhesion, cell phenotype and TNF-alpha release activity of such differentiated cells grown in hydrophobic teflon bags. Scanning electron microscopy showed that macrophages substantially adhered and spread on titanium-alloy surface throughout the culture period, whereas only a few dendritic cells were adherent. The phenotype of both cell types remained unchanged in the presence of the tested material. However, titanium-alloy stimulated the secretion of TNF-alpha by the macrophages of some donors. This model of culture may offer new insights into the biomaterial evaluation and may be useful for studying individual responses induced by biomaterials. PMID- 15348299 TI - Development of novel elastomer/methacrylate monomer soft lining materials. AB - An earlier study identified a formulation comprising a butadiene/styrene copolymer (PBS) gelled with ethyl hexyl methacrylate (5 + formulation) as a potential denture soft lining material. It had good mechanical properties but water uptake was high as a result of the presence of a separating agent. This study has compared the tensile and water absorption properties of four elastomers free from separating agent (three butadiene/styrene, HBS, EBS, SBS, and one isoprene/styrene, SIS) with those of PBS all using the 5 + formulation. HBS is emulsion polymerized; the others are solution polymerized. SIS5 + had the better tensile properties whereas HBS5 + had the lowest water uptake. All the other 5 + formulations had higher uptakes than PBS5 +, which is thought to be due to the presence of hydrophilic groups from the solution polymerization process. All materials showed some sign of oxidation. Emulsion polymerized elastomers are regarded as less suitable for medical uses than the solution polymerized alternatives. Of these, SIS5 + seems to be less prone to oxidation and has the better mechanical properties so was considered to be the most suitable material for further development. PMID- 15348300 TI - Interleukin-6 expression by cultured human endothelial cells in contact with carbon coated polyethylene terephthalate. AB - In order to evaluate if carbon coated polyethylene terephthalate (C-PET) could favor inflammatory reactions, the expression of interleukin-6 (IL-6) by cultured human umbilical vein endothelial cells was tested in vitro. The cultures were put in contact with C-PET for 1, 24, 48 and 72 h. The same cells cultured on tissue culture-treated polystyrene without biomaterials were tested as the negative control; the same cells incubated with LPS were the positive control. The level of IL-6 in the conditioned medium was tested by enzyme immunoassay; the mRNA expression was evaluated by RT-PCR with specific primers. The cultures incubated with C-PET produced non significantly different amount of IL-6 compared to the negative control and did not induce the expression of IL-6 specific mRNA. LPS induced a significantly higher release of IL-6 in the medium and the expression of mRNA after 24, 48 and 72 h. We conclude that C-PET does not stimulate the synthesis of IL-6 and therefore does not favor inflammatory reaction through the release of this cytokine. PMID- 15348301 TI - Compositional and morphological imaging of CO2 laser irradiated human teeth by low vacuum SEM, confocal laser scanning microscopy and atomic force microscopy. AB - Enamel and dentin of human teeth irradiated by CO2 laser were investigated by confocal laser scanning microscopy (CLSM), low vacuum scanning electron microscopy (WET-SEM) and atomic force microscopy (AFM). Optical tomographic imaging by CLSM, compositional imaging based on atomic number effect of reflected electrons by WET-SEM, high resolution observation of surface morphology by AFM were done for both the irradiated and non-irradiated area of the same specimen throughout. The crystals of about 50 microm length and the bright spots were observed by CLSM at the bottom of the cavity induced by laser irradiation. They turned out from the observation by WET-SEM as the acicular crystals with the cross section of an irregularly hexagonal shape situated parallel and perpendicular, respectively, to the inner surface of the cavity. The thickness of the thermally deteriorated zone of the cavity was about 25 microm. The crystals unidirectionally grown up to the size of several hundreds nm were also observed by AFM, while the apatite crystallites of 50-150 nm were recognized all over in non-irradiated area. All the results suggest that after instantaneous melting at the surface of teeth by CO2 laser shot the crystals of calcium phosphate were recrystalized and grown to a large size. The compositional imaging in addition to morphological observation was useful to obtain the information of the change in materials induced by laser irradiation. PMID- 15348302 TI - The chorioallantoic membrane of the chick embryo as a simple model for the study of the angiogenic and inflammatory response to biomaterials. AB - Angiogenesis is essential in wound healing and a common feature in chronic inflammation which is crucially involved in the biological response to biomaterials. A useful system to evaluate the angiogenic activity and the inflammatory potency of various agents is the chorioallantoic membrane (CAM) of the chick embryo. Here we examined its response to different biomaterials. Smooth materials such as PVC or the polyurethane Tecoflex either unmodified or modified by an OH- or N(CH(3))(3)(+)-end group (HEMA or MAPTAC) inhibited angiogenesis and did not induce the formation of granulation tissue. The anti-angiogenic effects of PVC, Tecoflex and its HEMA modification, however, were only seen at an early stage of development. In contrast, the MAPTAC modified Tecoflex inhibited angiogenesis over the whole time. Rough materials, e.g. filter paper or a collagen/elastin membrane, stimulated angiogenesis and induced the formation of inflammatory tissue. Histological analysis revealed that the filter material was homogeneously populated with cells consisiting mainly of macrophages, fibroblasts and endothelial cells. The collagen/elastin membrane was only partially infiltrated with cells. Among those also clusters of granulocytes were present pointing to an acute inflammatory process. These data show that the angiogenic activity and inflammatory response of biomaterials strongly depend on the chemical composition and the physical structure of the material. The CAM assay appears to be a useful tool for studying biocompatibility. PMID- 15348303 TI - Light scattering experiments on aqueous solutions of selected cellulose ethers: contribution to the study of polymer-mineral interactions in a new injectable biomaterial. AB - Hydroxypropylmethylcellulose (HPMC) is used as a ligand for a bioactive calcium phosphate ceramic (the filler) in a ready-to-use injectable sterilized biomaterial for bone and dental surgery. Light scattering experiments were usually used to study high water-soluble polymers and to determine the basic macromolecular parameters. In order to gain a deeper understanding of polymer/mineral interactions in this type of material, we have investigated the effect of divalent and trivalent ions (Ca(2+), PO(4)(3-)) and steam sterilization on dilute solutions of HPMC and hydroxyethylcellulose (HEC). The sterilization process may cause some degradation of HEC taking into account its high molecular weight and some rigidity of the polymer chain. Moreover, in the case of HPMC, the changes in the conformations rather than degradation process are supposed. These effects of degradation and flocculation are strengthened in alkaline medium. Experimental data suggested the formation of chelate complexes between Ca(2+) and HPMC which improve its affinity to the mineral blend and consolidate the injectable biomaterial even in the case of its hydration by biological fluid. PMID- 15348304 TI - Replacing the nucleus pulposus of the intervertebral disk: prediction of suitable properties of a replacement material using finite element analysis. AB - An axisymmetric finite element model of a human lumbar disk was developed to investigate the properties required of an implant to replace the nucleus pulposus. In the intact disk, the nucleus was modeled as a fluid, and the annulus as an elastic solid. The Young's modulus of the annulus was determined empirically by matching model predictions to experimental results. The model was checked for sensitivity to the input parameter values and found to give reasonable behavior. The model predicted that removal of the nucleus would change the response of the annulus to compression. This prediction was consistent with experimental results, thus validating the model. Implants to fill the cavity produced by nucleus removal were modeled as elastic solids. The Poisson's ratio was fixed at 0.49, and the Young's modulus was varied from 0.5 to 100 MPa. Two sizes of implant were considered: full size (filling the cavity) and small size (smaller than the cavity). The model predicted that a full size implant would reverse the changes to annulus behavior, but a smaller implant would not. By comparing the stress distribution in the annulus, the ideal Young's modulus was predicted to be approximately 3 MPa. These predictions have implications for current nucleus implant designs. PMID- 15348305 TI - X-ray diffraction characterization of dental gold alloy-ceramic interfaces. AB - X-ray diffraction (XRD) was employed to study dental alloy-ceramic interfaces. A Au-Pd-In alloy, which requires oxidation before porcelain firing, and a Au-Pt-Pd In alloy, which does not require oxidation before porcelain firing, were selected in this study. Alloy specimens were centrifugally cast. Specimen surfaces were metallographically polished through 0.05 microm Al2O3 slurries. A thin layer (< 50 microm) of a dental opaque porcelain was fired on the alloy surfaces with and without initial oxidation. XRD was conducted at room temperature on four types of alloy specimens: polished, oxidized, porcelain fired after alloy oxidation, and porcelain fired without initial alloy oxidation. XRD was also performed on fired opaque porcelain without an alloy substrate. The detection of prominent gold solid solution peaks from alloy-ceramic specimens indicated that the incident X ray beam reached the alloy-ceramic interface. In2O3 and beta-Ga2O3 were identified on the oxidized Au-Pd-In alloy, while In2O3 and SnO2 were detected on the oxidized Au-Pt-Pd-In alloy. Preferred orientation was observed for all the oxides formed on the alloys. Minimum lattice parameter changes (<1%) for the gold solid solutions were observed for both alloys before and after oxidation and porcelain firing. Leucite (KAlSi2O6, TiO2, ZrO2 and SnO2 were detected on the fired opaque porcelain. For both alloys, no additional oxides were identified at the metal-ceramic interfaces beyond those present in the oxidized alloys and the opaque porcelain. Similar results were obtained from alloy-ceramic interfaces where there was no prior alloy oxidation. The results indicate the critical role of alloy surface oxides in metal-ceramic bonding and support the chemical bonding mechanism for porcelain adherence. PMID- 15348306 TI - Titanium powder sintering for preparation of a porous functionally graded material destined for orthopaedic implants. AB - This work focuses on basic research into a P/M processed, porous-surfaced and functionally graded material (FGM) destined for a permanent skeletal replacement implant with improved structural compatibility. Based on a perpendicular gradient in porosity the Young's modulus of the material is adapted to the elastic properties of bone in order to prevent stress shielding effects and to provide better long-term performance of the implant-bone system. Using coarse Ti particle fractions the sintering process was accelerated by silicon-assisted liquid-phase sintering (LPS) resulting in a substantial improvement of the neck geometry. A novel evaluation for the strength of the sinter contacts was proposed. The Young's modulus of uniform non-graded stacks ranged from 5 to 80 GPa as determined by ultrasound velocity measurements. Thus, the typical range for cortical bone (10-29 GPa) was covered. The magnitude of the Poisson's ratio proved to be distinctly dependent on the porosity. Specimens with porosity gradients were successfully fabricated and characterized using quantitative description of the microstructural geometry and acoustic microscopy. PMID- 15348307 TI - Controlled release of salicylic acid from poly(D,L-Lactide). AB - Poly(D,L-Lactide) of high molecular weight (Mv) was prepared by ring-opening bulk polymerization of D,L-Lactide and characterized in terms of Mv, melting point and swelling behavior in buffer solution. Samples of the polymers with low and high Mv (2000 and 22 000 respectively) loaded with various amounts of salicylic acid (SA) were immersed in a buffer solution and the release of SA was recorded. The results obtained showed that swelling of the poly(D,L-Lactide) samples obeyed Fick's law, especially for those with high molecular weight, where biodegradation proceeds slowly. The release of SA seemed to follow a simplified relationship which is linear with time, at least for the early stages of delivery. The extent of linearity is dependent on the content of the acidic SA, which probably accelerates decomposition of the high molecular weight products. PMID- 15348308 TI - Laser confocal microscopic study of pH profiles of synthetic absorbable fibers upon in vitro hydrolytic degradation. AB - The objective of this study was to develop a new and non-destructive technique to measure the interior pH of synthetic absorbable biomaterials. Such a measurement would provide the required experimental evidence for validating the postulated theory that the accumulation of acidic hydrolytic degradation products within the interior of aliphatic polyesters is responsible for the observed accelerated degradation of this class of absorbable biomaterials. This new technique used a laser scanning confocal microscope coupled with pH sensitive fluorescent dyes like Texas Red sulfonyl chloride. The capability of optical thin sectioning of a laser confocal microscope would permit a non-destructive examination of the interior of biomaterials. Poly-p-dioxanone suture fibers (PDSII) of size 2/0 were used as the model compound for this new technique. The pH values of the unhydrolyzed and partially hydrolyzed PDSII fibers were found to decrease with increasing depth from the fiber surface and reached as low as about 3.5 at 70 microm depth. The largest depth that an interior pH could be measured within absorbable biomaterials was determined by the opacity of the biomaterials, i.e. a higher depth for a less opaque material. The observed interior pH profiles were correlated to the unique morphologic structure of PDSII fibers. PMID- 15348309 TI - Comparison of the response of three human monocytic cell lines to challenge with polyethylene particles of known size and dose. AB - The response of three human monocytic cell lines (Monomac-1, U937 and THP-1) to challenge with polyethylene particles of known size and dose was evaluated. Particles with a mean size of 0.21, 0.49, 4.3, 7.2, and 88 microm were co cultured with the cells for 24 hours prior to the assessment of cell viability and production of the pro-inflammatory cytokines, IL-1beta, IL-6 and TNFalpha. Additionally, GM-CSF and prostaglandin E2 were measured in culture supernatants from particle stimulated U937 cells. All particle fractions were evaluated at particle volume (microm3) to cell number ratios of 100 : 1, 10 : 1, 1 : 1 and 0.1 : 1. None of the test fractions had any effect on cell viability. Only the response of the U937 cell line was demonstrated to be comparable to that of primary macrophages as determined in a previous study. Furthermore only particle volume to cell number ratios of 10 : 1 or greater consistently stimulated significantly enhanced levels of cytokine secretion with particles within the phagocytosable size range (0.1 to 10 microm) being the most biologically active. No response was observed when U937 macrophages were stimulated with the largest (88 microm) particles at any of the volume ratios tested in this study. These results suggest that the size and volume of polyethylene particles are critical factors in macrophage activation. In addition, the U937 cell line has been shown to be a suitable model for the in vitro study of macrophage-particle interactions. PMID- 15348310 TI - Evaluation of the effect of three calcium phosphate powders on osteoblast cells. AB - The aim of the present study was to assess the effect of three calcium phosphate powders entering in the composition of bone substitute materials on osteoblast cells activity. These powders were hydroxyapatite (HA) widely used as a biomaterial, nanocrystalline carbonate apatite (C A) very close to bone mineral crystals, and an experimental one: calcium phosphate cement-1 (CPC-1) composed of an amorphous Ca-P phase and brushite. The powders were physico-chemically characterized. The very reactive CPC-1 powder became transformed in cell culture medium: recrystallization of amorphous precursors and hydrolysis of brushite into poorly crystalline apatite occurred. Osteoblast-cells activity was evaluated: for low level of calcium phosphates (>100 microg/ml) CPC-1 enhanced proliferation and, to a lesser degree, differentiation on alkaline phosphatase activity. For 100 microg/ml of powders we observed a great alteration of biological activity of the osteoblasts: evaluation of proliferation indicated an inhibition for all samples, and a decrease of two differentiation markers: alkaline phosphatase activity and osteocalcin release were noticed, suggesting a down regulation due to the presence of large amount of mineral powder. PMID- 15348311 TI - Fatigue characteristics of ultra high molecular weight polyethylene with different molecular weight for implant material. AB - In order to investigate the effect of molecular weight on fatigue characteristics in the ultra high molecular weight polyethylene (UHMWPE), tension-tension fatigue tests of notched specimens were carried out in the present study. The effects of frequency and stress ratio on the fatigue characteristics were also investigated and fractography was discussed. The fatigue strength does not increase with increasing molecular weight. The fatigue strength might be influenced by the high degree of crystallinity in spite of the decreased tie molecule density in this study. Almost no effect of frequency on the number of cycles to failure can be observed. However, the higher the frequency, the higher the crack tip temperature. The effects of heat and strain rate on the fatigue strength must be considered in polymer materials. At a high stress ratio, the stress-number of cycles to failure (S-N) curves shift to high number cycles to failure side. Both stress amplitude and mean stress influence the fatigue life of UHMWPE. PMID- 15348312 TI - A study of histological responses from Ti-6Al-7Nb alloy dental implants with and without plasma-sprayed hydroxyapatite coating in dogs. AB - Titanium alloys are hoped to be used much more for applications as implant materials in the medical and dental fields because of their basic properties, such as biocompatibility, corrosion resistance and specific strength compared with other metallic implant materials. Thus, the Ti-6Al-7Nb alloy that has recently been developed for biomedical use, that is, primarily developed for orthopaedic use, is to be studied in this paper, for application in dental implants. The biocompatibility test in vivo was carried out in dogs and the osseointegration was verified through histological analysis of the samples of the Ti-6Al-7Nb alloy with and without hydroxyapatite coating that were inserted in the alveoli. Within the controlled conditions the samples did not show any toxic effects on the cells. PMID- 15348313 TI - Polishing and whitening properties of toothpaste containing hydroxyapatite. AB - Toothpastes containing hydroxyapatite (HA-toothpaste) have demonstrated efficacy in preventing caries, treating hypersensitivity and curing periodontal diseases, however, their effects on whitening teeth have not been investigated. In this study, the polishing and whitening properties of hydroxyapatite contained toothpastes were evaluated in a combined experimental and clinical study. The polishing properties were evaluated in artificial teeth by polishing with different toothpastes, while the brightening and whitening properties were examined in volunteers using two colorimeters with two specially made fiberscopes. The results revealed that the addition of hydroxyapatite to toothpaste did not alter its polishing properties, while it did result in a marked increase in tooth-whiting. It was also found that the brightening and whitening properties increased as the amount of hydroxyapatite in the toothpaste increased. It is concluded that toothpaste containing hydroxyapatite are effective at whitening tooth and that whitening was not due to their polishing effect on tooth surface. PMID- 15348314 TI - Role of HSP70i in regulation of biomaterial-induced activation of human monocytes derived macrophages in culture. AB - The functioning of an implant depends on the material properties and the wound healing process. The latter is led by an inflammatory reaction guided mainly by monocyte/macrophage activity. This in vitro study investigated human monocytes/macrophages in culture from 2 h to 10 days on silicone, polyurethane, teflon and TCPS. Analysis of cytokine release by ELISA showed that maturing macrophages have different capacities to produce cytokines TNFalpha, IL10, IL8 and GM-CSF. The long culture-mature macrophages on all polymers produced comparable low levels of TNFalpha, IL10 and IL8. Monocytes/macrophages on polyurethane and teflon, and those on silicone only in long culture-time produced high GM-CSF amounts, where as those on TCPS exhibited low levels of GM-CSF. FACS analysis revealed that HSP70i was highly inducible after short time culture yet this high level was maintained in long culture-mature macrophages on TCPS only, whereas on other polymers the mature macrophages showed a high reduction in HSP70i level, which demonstrated a high stress-response by cells on TCPS. Accordingly, CLSM-analysis revealed low nuclear NF-kappaB in cells on TCPS and high nuclear NF-kappaB in mature macrophages on silicone and polyurethane, showing a high cellular activation on the latter two polymers. This corresponded also to the high mitochondrial activity by XTT metabolism displayed by the mature macrophages on polyurethane >/= silicone > teflon > TCPS. These data show a correlation of (1) cytokines (TNFalpha, GM-CSF) and HSP70i, (2) NF-kappaB and HSP70i by monocytes/macrophages after contact with polymers. Thus, HSP70i might be a useful molecular candidate for exploring biomaterial-induced inflammatory reaction. PMID- 15348315 TI - Stability study of tetracyclines with respect to their use in slow release systems. AB - In the aim of optimizing implantable slow-release systems for the local delivery of antibiotics, the stability of tetracyclines was studied in water at 37 degrees C or under gamma irradiation. Four tetracyclines in their chlorhydrate form were chosen depending on their hydrophilic/hydrophobic balance. Their chemical stability was established by HPLC, and biological stability by bacteriological tests. It was shown that methacycline and doxycycline are stable in water for three days. Tetracycline and minocycline exhibit limited decomposition (less than 10%) under the same conditions. So, in vitro drug release for at most three days, appears to be possible. Besides, all four tetracyclines either in powdered form or included in a calcium phosphate matrix, kept their bacteriological activity after gamma irradiation at 32.4 kGr. Consequently, the in vivo study of these implantable slow drug release systems, can be carried out. PMID- 15348316 TI - Functionalization of silicone rubber for the covalent immobilization of fibronectin. AB - Surface modification techniques were employed in order to provide functionalized silicone rubber with enhanced cytocompatibility. Acrylic acid (AAc), methacrylic acid (MAAc) and glycidylmethacrylate (GMA) were graft-co-polymerized onto the surface of silicone induced by an argon plasma and thermal initiation. The polymerizations were carried out in solution, in the case of acrylic acid a vapor phase graft-co-polymerization subsequent to argon plasma activation was carried out as well. Human fibronectin (hFn), which acts as a cell adhesion mediator for fibroblasts, was immobilized by making use of the generated carboxylic or epoxy groups, respectively. Surface analysis was accomplished by means of X-ray photoelectron spectroscopy (XPS), infrared spectroscopy in attenuated total reflection mode (IR-ATR), scanning electron microscopy (SEM), atomic force microscopy (AFM) and dynamic contact angle measurements using the Wilhelmy-plate method. The amount of immobilized active hFn was semiquantified by enzyme-linked immunosorbent assay (ELISA) using a structure-specific antibody against the cell binding domain of hFn. In vitro testing showed a remarkable difference between surfaces exposing adsorbed-only and surfaces with covalently immobilized hFn. PMID- 15348317 TI - Dextran-magnetite complex: conformation of dextran chains and stability of solution. AB - Three kinds of dextran-magnetite (DM) complexes were prepared using alkali treated dextrans with molecular weights of 1900, 4200 and 9600, respectively. The number of dextrans binding to a magnetite core was determined. The number was proportional to the area of core surface and the area occupied by a dextran was 2.5 nm2 for molecular weight of 1900, 2.8 for 4200 and 3.8 for 9600. The binding of dextrans to core may be conditioned by the conformation of dextran chains in water (possibly by the steric hindrance between dextrans covering core). Stability of the DM solution was examined at 80 degrees C. Aggregation and/or precipitation of DM particles were observed within two weeks. The stability of DM solution was found to increase with increasing molecular weight of dextran. The dissociation of dextran from the core may cause the aggregation and subsequent precipitation of DM particles (the dissociation constant at 20 degrees C, 3.7 x 10(-6) for a molecular weight of 1900 and 5.4 x 10(-7) for 9600). PMID- 15348318 TI - A percutaneous device as model to study the in vivo performance of implantable amperometric glucose sensors. AB - Glucose kinetics were investigated in subcutaneous tissue of rabbits, in which a percutaneous device was implanted. The device was used for collection of tissue fluid and as carrier of an amperometric glucose sensor. Changes in glycaemia were reflected in subcutaneous tissue fluid. However, a limited number of responses of the implanted sensors were observed. Histologic evaluation showed thin fibrous capsules surrounding the implants. Accumulations of inflammatory cells were observed inside the subcutaneous chamber. The experiments again showed that changes in blood glucose concentration can be measured in subcutaneous tissue fluid collected with a percutaneous device. Nevertheless, implanted glucose sensors could not reliably monitor these changes. Supported by our histological observations and sufficient in vitro performance, we suppose that the cellular reaction to the sensor plays an important role in this poor in vivo performance. In combination with adsorption of tissue fluid proteins, this results in a reversible deactivation of implanted sensors. The exact mechanisms involved in this process are currently unknown and need further investigation. PMID- 15348319 TI - Formation of macropores in calcium phosphate cement implants. AB - A calcium phosphate cement (CPC) was shown to harden at ambient temperatures and form hydroxyapatite as the only end-product. Animal study results showed that CPC resorbed slowly and was replaced by new bone. For some clinical applications, it would be desirable to have macropores built into the CPC implant to obtain a more rapid resorption and concomitant osseointegration of the implant. The present study investigated the feasibility of a new method for producing macropores in CPC. Sucrose granules, NaHCO3, and Na2HPO4 were sieved to obtain particle sizes in the range of 125 microm to 250 microm. The following mixtures of CPC powder (an equimolar mixture of tetracalcium phosphate, Ca4(PO4)2O, and dicalcium phosphate anhydrous, CaHPO4) and one of the above additive granules were prepared: control-no additive; mixture A-0.25 mass fraction of sucrose; mixture B 0.25 mass fraction of NaHCO3; mixture C-0.25 mass fraction of Na2HPO4, and mixture D-0.33 mass fraction of Na2HPO4. Cement samples were prepared by mixing 0.3 g of the above mixtures with 0.075 ml of the cement liquid (1 mol/l Na2HPO4). After hardening, the specimens were placed in water for 20 h at about 60 degrees C to completely dissolve the additive crystals. Well-formed macropores in the shapes of the entrapped crystals were observed by scanning electron microscope (SEM). The macroporosities (mean+/-standard deviation; n = 6) expressed as volume fraction in % were 0, 18.9 +/- 1.7, 26.9 +/- 1.6, 38.3 +/- 4.4 and 50.3 +/- 2.7 for the control, A, B, C and D, respectively. The diametral tensile strengths (mean+/-standard deviation; n = 3) expressed in MPa were 10.1 +/- 0.7, 3.7 +/- 0.3, 2.4 +/- 0.2, 1.5 +/- 0.5 and 0.4 +/- 0.1, respectively, for the five groups. The results showed that macropores can readily be formed in CPC implants with the use of water-soluble crystals. The mechanical strength of CPC decreased with increasing macroporosity. PMID- 15348320 TI - The influence of acetabular cup angle on the wear of "BIOLOX Forte" alumina ceramic bearing couples in a hip joint simulator. AB - The wear of "Biolox Forte" alumina ceramic bearing couples has been investigated at two different acetabular cup angles in a physiological hip joint simulator. All cups were set in the anatomical position of 45 degrees inclination in the M/L plane for the first two million cycles and then four of the six cups were re aligned to 60 degrees for a further three million cycles. A "running-in" wear of 0.14 mm3 per million cycles was observed for the first million cycles, after which a steady state wear rate of 0.05 mm3 per million cycles was observed. Increasing the acetabular cup angle to 60 degrees did not significantly affect the wear rate. PMID- 15348321 TI - Corrosion behavior of PIRAC nitrided Ti-6Al-4V surgical alloy. AB - Hard titanium nitride (TiN) coatings were obtained on the surface of Ti-6Al-4V alloy using an original PIRAC nitriding method, based on annealing the samples under a low pressure of monatomic nitrogen created by selective diffusion of N from the atmosphere. PIRAC nitrided samples exhibited excellent corrosion resistance in Ringer's solution in both potentiodynamic and potentiostatic tests. The anodic current and metal ion release rate of PIRAC nitrided Ti-6Al-4V at the typical corrosion potential values were significantly lower than those of the untreated alloy. This, together with the excellent adhesion and high wear resistance of the TiN coatings, makes PIRAC nitriding an attractive surface treatment for Ti-6Al-4V alloy surgical implants. PMID- 15348322 TI - Characterization of the kinetic behavior of resin modified glass-ionomer cements by DSC, TMA and ultrasonic wave propagation. AB - In this study the isothermal kinetic behavior of two resin modified glass ionomer cements (RMGIC) and a dental composite have been compared by differential scanning calorimetry (DSC) and thermo-mechanical analysis (TMA). The simultaneous evolution of the multiple reactions occurring in RMGIC has been analyzed not only by DSC and TMA but also by ultrasonic wave propagation using the pulse-echo technique. The propagation of ultrasonic waves, acting as a dynamic mechanical deformation at high frequencies, is proportional to the longitudinal bulk moduli of the material and may be used to measure the changes of mechanical properties induced by a chemical reaction as occurs in RMGIC. TMA and ultrasonic analysis have been used to monitor the acid-base reaction of RMGIC in dark conditions. Moreover an RMGIC presenting a double reactive mechanism in dark conditions, a thermally activated radical polymerization and an acid-base reaction are studied using these experimental techniques. Finally DSC and TMA results obtained during photopolymerization of an RMGIC and of a dental composite have been compared. PMID- 15348323 TI - Studies on polyurethane potting compound based on HDI-TMP adduct for fabrication of haemodialyzer. AB - Polyurethane potting compounds based on hexamethylene diisocyanate-trimethylol propane (HDI-TMP) adduct (Component "A") and polypropylene glycol, polyethylene glycol and castor oil (Component "B") were prepared as potential compounds for the fabrication of haemodialyzer. The setting characteristics of the potting compounds having isocyanate index 2.0 are better than those compounds having 1.35. The aging stability of PEG and PPG based potting compounds are poorer than those of castor oil based potting compounds. Appreciable hydrolytic, oxidative and chemical stability could be observed with castor oil based potting compounds of HDI-TMP adduct. PMID- 15348324 TI - Effect of powder characteristics on the sinterability of hydroxyapatite powders. AB - The effect of different sintering conditions on the sintered density and microstructure of two different hydroxyapatite (HA) powders was examined. The powder characteristics of a laboratory synthesized HA powder (Lab HA) were low crystallinity, a bimodal particle size distribution, a median particle size of 22 microm and a high specific surface area (SSA) of 63 m2/g. By contrast, a commercial calcined HA (commercial HA) was crystalline and had a median particle size of 5 microm and a low SSA of 16 m2/g. The different powder characteristics affected the compactability and the sinterability of the two HA powders. Lab HA did not compact as efficiently as commercial HA, resulting in a lower green density, but the onset of sintering of powder compacts of the former was approximately 150 degrees C lower than the later. The effect of compaction pressure, sintering temperature, time and heating rate on the sintered densities of the two materials was studied. Varying all these sintering conditions significantly affected the sintered density of commercial HA, whereas the sintered density of Lab HA was only affected significantly by increasing the sintering temperature. The Vickers hardness, Hv, of Lab HA was greater than commercial HA for low sintering temperatures, below 1200 degrees C, whereas for higher sintering temperatures the commercial HA produced ceramics with greater values of hardness. These trends can be related to the sinterability of the two materials. PMID- 15348325 TI - Acetal-functionalized polymer particles useful for immunoassays. III: preparation of latex-protein complexes and their applications. AB - Monodisperse polymer colloids with dimethyl and diethyl acetal functionalities synthesized by a two-step emulsion polymerization process were chosen as the polymeric support to carry out covalent coupling with the antibody IgG anti C reactive protein, and to test the utility of the latex-protein complexes formed in immunoassays with the specific CRP antigen. More than the 80% of the initially linked protein was covalently coupled in all of the latexes. The agglutination reaction was followed by turbidimetry. With the aim of analyzing the effect of some of the variables of the immunological reaction, the reaction time, the particle concentration and the coverage degree of protein in the complexes were varied. PMID- 15348326 TI - Calcining influence on the powder properties of hydroxyapatite. AB - The effect of different calcination temperatures on the powder characteristics and the sintered density of synthetic hydroxyapatite (HA) powders, produced using two different processing routes, was examined. Powders were produced by either drying, milling and sieving an as-precipitated HA or by spray-drying a slurry of precipitated HA. Calcining the two powders at temperatures between 400 and 1000 degrees C did not significantly affect the powder particle size. The specific surface areas of the two powders, however, were reduced from 70-80 m2/g for a calcination temperature of 400 degrees C to approximately 5-7 m2/g for 1000 degrees C. Analysis of the surfaces of the HA powders using scanning electron microscopy (SEM) illustrated the coarsening and subsequent sintering of the sub micron crystallites that constitute a powder particle as the calcination temperature increased, corresponding to the decrease in surface area of the powders. The sintered densities of the final ceramics were not significantly affected by calcining the powders. Microhardness measurements of ceramics prepared from powders calcined at different temperatures showed no significant variations with calcination temperature or powder processing method. The results of this study have illustrated that for applications where HA may be used in powder form, for example in plasma-spraying and for the production of HA-polymer composites, calcining the HA will significantly affect the powder properties, namely the surface area and morphology of the powders. For applications requiring HA in a dense ceramic form, for example as granules or blocks, calcining the powders does not significantly affect the properties of the final ceramic. PMID- 15348328 TI - Silk fibroin-polyurethane scaffolds for tissue engineering. AB - Silk fibroin (SF) is a highly promising protein for its surface and structural properties, associated with a good bio- and hemo-compatibility. However, its mechanical properties and architecture cannot be easily tailored to meet the requirements of specific applications. In this work, SF was used to modify the surface properties of polyurethanes (PUs), thus obtaining 2D and 3D scaffolds for tissue regeneration. PUs were chosen for their well known advantageous properties and versatility; they can be obtained either as 2D (films) or 3D (foams) substrates. Films of a medical-grade poly-carbonate-urethane were prepared by solvent casting; PU foams were purposely designed and prepared with a morphology (porosity and cell size) adequate for cell growth. PU substrates were coated with fibroin by a dipping technique. To stabilize the coating layer, a conformational change of the protein from the alpha-form (water soluble) to the beta-form (not water soluble) was induced. Novel methodology in UV spectroscopy were developed for quantitatively analyzing the SF-concentration in dilute solutions. Pure fibroin was used as standard, as an alternative to the commonly used albumin, allowing real concentration values to be obtained. SF-coatings showed good stability in physiological-like conditions. A treatment with methanol further stabilized the coating. Preliminary results with human fibroblasts indicated that SF coating promote cell adhesion and growth, suggesting that SF-modified PUs appear to be suitable scaffolds for tissue engineering applications. PMID- 15348329 TI - Manufacture and evaluation of bioactive and biodegradable materials and scaffolds for tissue engineering. AB - For tissue regeneration and tissue engineering applications, a number of bioactive and biodegradable composites, either porous or non-porous, were fabricated. The newly developed materials included tricalcium phosphate reinforced polyhydroxybutyrate and its copolymer, poorly crystallized hydroxyapatite reinforced chitin, and plasma sprayed hydroxyapatite reinforced poly(L-lactic acid). It was shown that these new materials could be successfully produced using the manufacturing techniques adopted. In vitro experiments revealed that the incorporation of bioceramic particles in biodegradable polymers rendered the composites bioactive and significantly improved the ability of composites to induce the formation of bone-like apatite on their surfaces. Degradation of composite scaffolds in simulated body fluid was observed and could be due to the simultaneous degradation of polymer matrix and dissolution of bioceramic particles. PMID- 15348330 TI - Chitosan-based hydrogels: synthesis and characterization. AB - Chitosan (CHI) is a polysaccharide of beta-1,4-linked 2-amino-2-deoxy-D glucopyranose derived by N-deacetylation of chitin in aqueous alkaline medium. The shells of crustaceans such as crabs, shrimp, and lobster are the current source of chitosan. It is known to be non-toxic, odourless, biocompatible in animal tissues and enzymatically biodegradable. For these reasons much research interest has been paid to its biomedical, ecological, and industrial applications over the past decade. However, its rigid crystalline structure, poor solubility in organic solvents and poor processability have limited its use. To broadening its range of applications, a growth research effort has been devoted to explore ways of modifying Chitosan. Here it has been reported on the synthesis of new hydrogels, obtained by self-curing chitosan with acrylic acid (AA) and methyl acrylate (MA). The hydrogels were characterized by FTIR, swelling and rheological analysis. The results of this study showed that the swelling and mechanical properties of chitosan are highly improved by the presence of poly acrylate. The swelling degree of these materials does not depend upon the ratio MA/AA. It is possible to improve and modulate the mechanical properties of the hydrogels by changing the relative MA/AA ratio. PMID- 15348331 TI - Phosphorylcholine-based polymer coatings for stent drug delivery. AB - Phosphorylcholine-based polymers have been used commercially to improve the biocompatibility of coronary stents. In this study, one particular polymer is assessed for its suitability as a drug delivery vehicle. Membranes of the material are characterized in terms of water content and molecular weight cut off, and the presence of hydrophilic and hydrophobic domains investigated by use of the hydrophobic probe pyrene. The in vitro loading and elution of a variety of drugs was assessed using stents coated with the polymer. The rate of a drug's release was shown not to be simply a function of its water solubility, but rather more closely related to the drug oil/water partition coefficient. This finding was explained in terms of the more hydrophobic drugs partitioning into, and interacting with, the hydrophobic domains of the polymer coating. The suitability of the coated stent as a drug delivery vehicle was assessed in vivo using a radiolabeled analog of one of the more rapidly eluting drugs, angiopeptin. Autoradiography showed that the drug was released locally to the wall of the stented artery, and could be detected up to 28 days after implantation. PMID- 15348332 TI - Biomimetic coatings functionalized with adhesion peptides for dental implants. AB - A complete biological integration into the surrounding tissues (bone, gingiva) is a critical step for clinical success of a dental implant. In this work biomimetic coatings consisting either of collagen type I (for the gingiva region) and hydroxyapatite (HAP) or mineralized collagen (for the bone interface) have been developed as suitable surfaces regarding the interfaces. Additionally, using these biomimetic coatings as a matrix, adhesion peptides were bound to further increase the specificity of titanium implant surfaces. To enhance cell attachment in the gingiva region, a linear adhesion peptide developed from a laminin sequence (TWYKIAFQRNRK) was bound to collagen, whereas for the bone interface, a cyclic RGD peptide was bound to HAP and mineralized collagen using adequate anchor systems. The biological potential of these coatings deduced from cell attachment experiments with HaCaT human keratinocytes and MC3T3-E1 mouse osteoblasts showed the best results for collagen and laminin sequence coating for the gingiva region and mineralized collagen and RGD peptide coatings for regions with bone contact. Our concept opens promising approaches to improve the biological integration of dental implants. PMID- 15348333 TI - Polyacrylamide hydrogels and semi-interpenetrating networks (IPNs) with poly(N isopropylacrylamide): mechanical properties by measure of compressive elastic modulus. AB - Semi-IPN hydrogels (based on cross-linked polyacrylamide having poly(N isopropylacrylamide) (PN1PAAm) inside) were synthesized and their properties, such as swelling ratio and compressive elastic moduli, were studied at several temperatures. Equilibrium swelling ratios of semi-IPN markedly decreased due to the presence of less hydrophilic PNIPAAm chains. The semi-IPN presented greater elastic modulus when compared to the cross-linked PAAm hydrogel. The effect was explained as being an additional contribution of the PNIPAAm chains, which collapsed around the PAAm networks, to the elastic modulus. It was pointed out that the PAAm networks support the collapsed chains. According to the results presented in this work, semi-IPN hydrogels present better mechanical properties than the PAAm hydrogel, mainly when the PNIPAAm chains are in a collapsed state. PMID- 15348334 TI - Tensile mechanical properties of polyacetal after one and six months' immersion in Ringer's solution. AB - The engineering polymer polyacetal (also known as polyoxymethylene) has a long history of clinical use including in the femoral component of the Freeman all polymer knee replacement. The polymer is a promising material for novel applications including the femoral component of hip resurfacing prostheses. However, no work following the effect on mechanical properties of extended immersion in the physiological environment has been reported. Samples of five grades of polyacetal were obtained and tested on an MTS Bionix 858 at 37 degrees C in physiological saline. Three groups of specimens were tested: "as received"; immersed in Ringer's solution at 37 degrees C for one month; immersed in Ringer's solution at 37 degrees C for six months'. While there were negligible reductions in the ultimate tensile strengths, the Young's moduli of all polymer grades showed significant reductions (p < 0.01) of up to 19.5% following one months' immersion. There were negligible changes in ultimate tensile strength and stiffness between one and six months' immersion, however some polymers showed a small, but significant (p < 0.05) increase in stiffness. The percentage elongation and true stress for plastic instability showed negligible changes following immersion for one and six months'. These results encourage the use of polyacetal in load-bearing orthopaedic applications. PMID- 15348335 TI - Tensile properties of a bone cement containing non-ionic contrast media. AB - The addition of contrast media such as BaSO4 or ZrO2 to bone cement has adverse effects in joint replacements, including third body wear and particle-induced bone resorption. Ground PMMA containing particles of the non-ionic water-soluble iodine-based X-ray contrast media, iohexol (IHX) and iodixanol (IDX), has, in bone tissue culture, shown less bone resorption than commercial cements. These water-soluble non-ceramic contrast media may change the mechanical properties of acrylic bone cement. The static mechanical properties of bone cement containing either IHX or IDX have been investigated. There was no significant difference in ultimate stress between Palacos R (with 15.0 wt % of ZrO2) and plain cement with 8.0 wt % of IHX or IDX with mass median diameter (MMD) of 15.0 or 16.0 microm, while strain to failure was higher for the latter (p < 0.02). The larger particles (15.0 or 16.0 microm) gave significantly higher (p < 0.001) ultimate tensile strengths and strains to failure than smaller sizes (2.4 or 3.6 microm). Decreasing the amount of IHX from 10.0 wt % to 6.0 wt % gave a higher ultimate tensile strength (p < 0.001) and strain to failure (p < 0.02). Scanning electron microscopy (SEM) showed the smaller contrast media particles attached to the surface of the polymer beads, which may prevent areas of the acrylate bead surface from participating in the polymerization. In conclusion, the mechanical properties of bone cement were influenced by the size and amount of contrast medium particles. By choosing the appropriate amount and size of particles of water-soluble non-ionic contrast media the mechanical properties of the new radio opaque bone cement can be optimized, thus reaching and surpassing given regulatory standards. PMID- 15348336 TI - Development of bioactive PMMA-based cement by modification with alkoxysilane and calcium salt. AB - Poly (methyl methacylate) (PMMA) bone cement is one of the popular bone-repairing materials for fixation of artificial hip joints. Significant problems on the PMMA bone cement are caused by loosening at the interface between bone and the cement, since the cement does not show bone-bonding, i.e. bioactivity. Development of PMMA bone cement capable of bone-bonding has been therefore long desired. The prerequisite for an artificial material to show bone-bonding is the formation of a biologically active bone-like apatite layer on its surface when implanted in the body. The same type of apatite formation can be observed on bioactive materials even in a simulated body fluid (Kokubo solution) with ion concentrations nearly equal to those of human blood plasma. Fundamental researches for bioactive glasses and glass-ceramics revealed that the apatite deposition is initiated by release of Ca2+ ions from the material into the body fluid, and by catalytic effect of Si-OH groups formed on the surface of the material. These findings lead an idea that novel bioactive cement can be designed by incorporation of Si-OH groups and Ca2+ ion into PMMA bone cement. In the present study, PMMA bone cement is modified with 20 mass % of various kinds of alkoxysilanes and calcium salts, and its apatite-forming ability was evaluated in Kokubo solution. The apatite formation was observed on the surface of the modified cements containing 20 mass % of CaCl2, irrespective of the kind of the examined alkoxysilane. On the other hand, the apatite formation was observed on the cement containing CaCl2, Ca(CH3COO)2 or Ca(OH)2, but not on the cement containing CaCO3 or beta-Ca3(PO4)2, even when the cement contains 3 methacryloxypropyltrimethoxysilane (MPS). The results indicate that modification with alkoxysilane and calcium salts showing high water-solubility is effective for providing PMMA bone cement with bioactivity. PMID- 15348337 TI - The influence of hydroxyapatite: zinc oxide ratio on the setting behavior and mechanical properties of polyalkenoate cements. AB - The influence of hydroxyapatite (HA) content on the setting behavior and mechanical properties of hydroxyapatite-zinc oxide-poly(acrylic acid) (HA-ZnO PAA) composite cements were investigated as a function of HA content. The working time increased with HA content up to 45 wt % HA and then decreased whilst the setting time increased with increasing HA content. Mechanical properties were determined after storage in water at 37 degrees C for 1, 7 and 28 days. Young's moduli and compressive strength go through a maximum at approximately 30 and 45 wt % HA. Young's modulus increases with time, which is consistent with an ongoing crosslinking reaction. PMID- 15348338 TI - Rheological properties of an apatitic bone cement during initial setting. AB - One scientific and technological aspect of main importance to the medical profession is to develop injectable calcium phosphate cements (CPCs) to be used through minimally invasive surgery techniques with still suitable mechanical and biodegradable properties. The objective of this research was to study the influence of several technological factors on the injectability of CPCs. This was performed by studying the rheological behavior of the cement pastes during their initial setting. Cement rheology was approached by looking at the creep response of apatitic cements as a function of the shear stress, the liquid-to-solid (L/S) ratio, the temperature and the addition of organic admixtures. Results showed creep experiments to be a finer method to detect characteristic setting times than other established subjective procedures. However, of all transition times detected none but the dough time seems to be of relevant importance when injectability of cement is concerned. Creep experiments also showed that the addition of organic admixtures such as citric acid increased injectability by retarding the hydration time. PMID- 15348339 TI - Effect of filler type on the mechanical properties of self-reinforced polylactide calcium phosphate composites. AB - Bioabsorbable polymers are of interest as internal fracture fixation devices. Self-reinforcement has been developed to improve the mechanical properties of the material and the addition of calcium phosphate fillers improves the bioactivity. Composite plates, produced by compression molding preimpregnated sheets of polylactide fibers coated in a polylactide matrix have been degraded in simulated body fluid for up to 12 weeks. Some samples also contained hydroxyapatite or tricalcium phosphate filler particles. Degradation was measured by monitoring the water uptake and mass decrease of the samples, as well as carrying out four point bend tests to assess the mechanical properties of the material. By 12 weeks, it was found that the unfilled samples absorbed more water and showed greater mass loss than the samples containing calcium phosphate fillers. Also, the flexural modulus and yield stress decreased significantly at week 12 for the unfilled samples. Adding hydroxyapatite (HA) or tricalcium phosphate (TCP) to the composite increased the flexural modulus and yield strength to values within the range of those reported for cortical bone and these values were maintained over the 12-week period. PMID- 15348340 TI - Bioactive response of Ag-doped tape cast Bioglass 45S5 following heat treatment. AB - The suitability of Bioglass 45S5 to the tape casting process and the ability of the glass to retain in vitro bioactivity following heat treatment to increase strength has been established. In this research, tape cast Bioglass was doped with silver prior to heat treatment in an effort to impart antimicrobial properties. The effect of initial dopant concentration and processing temperature was investigated. FTIR confirmed the presence of well established hydroxyapatite (HA) surface layers on Ag-doped tape cast Bioglass following heat treatment and simulated body fluid (SBF) immersion. Solution analysis revealed the release of silver ion in concentrations similar to those reported in the literature to have antimicrobial effects. Additionally, mercury porosimetry was used to determine that silver enhanced the densification process relative to that of undoped tape cast Bioglass. Ag-doped tape cast and sintered Bioglass (TCSBG) appears promising for use in situations requiring rapid surface HA formation and antimicrobial properties. PMID- 15348341 TI - Aligned bioactive mesoporous silica coatings for implants. AB - Ongoing research is reported aimed at preparing mesoporous silica coatings on various substrates for medical applications by a biomimetic approach (self assembling of organic/inorganic sol-gel systems into ordered structures). Tetraethylorthosilicate (TEOS) was selected as the silica precursor, and amphiphilic triblock copolymers poly(ethylene oxide)-poly(propylene oxide) poly(ethylene oxide), and the cationic surfactant cetyltrimethyl ammonium chloride (CTAC), as structure-directing agents. The mesochannels diameter could be adjusted by changing the directing agent, and a preferred alignment of the mesostructure was observed independently of the used substrate (glass, silicon, Ti or Ti6Al4V). Three different treatments (thermocalcination, photocalcination, and solvent extraction) have been also studied to remove the organic templates, of which photocalcination showed to be the most versatile. When soaked in a simulated body fluid, mesoporous silica coatings induced apatite formation after seven days. PMID- 15348342 TI - Optimizing a hydroxyapatite/tricalcium-phosphate ceramic as a bone graft extender for impaction grafting. AB - The mechanical properties of morsellized bone allografts and synthetic hydroxapatite/tricalcium-phosphate (HA/TCP) ceramic extender materials for the use in impaction grafting revision hip surgery were investigated using two test methods: a basic compression test and an endurance test in an in-vitro model of an impaction grafted femur. Formalin fixed ovine bone graft was identified as mechanically similar to fresh human bone and thus suitable as an experimental material for in-vitro testing. For 1 : 1 volumetric mixes of bone allograft and synthetic extender, the granular ceramic's properties were varied in porosity, chemical composition, sintering temperature and particle size. Initial mechanical stability, a crucial prerequisite for clinical success in impaction grafting, was increased for all bone/extender mixes. A high porosity, tricalcium-phosphate rich ceramic of medium particle size and sintered at high temperatures was recognized as an optimized extender material for impaction grafting balancing the mechanical and biological demands. Using the extender without bone graft as a pure replacement is not recommended. PMID- 15348343 TI - Low cycle fatigue behavior of Ti6Al4V thermochemically nitrided for its use in hip prostheses. AB - Titanium and its alloys have many attractive properties including high specific strength, low density, and excellent corrosion resistance. Besides, titanium and the Ti6Al4V alloy have long been recognized as materials with high biocompatibility. These properties have led to the use of these materials in biomedical applications. Despite these advantages, the lack of good wear resistance makes difficult the use of titanium and Ti6Al4V in some biomedical applications, like articulating components of prostheses. Some surface treatments are available in order to correct these problems, like thermal surface treatment by means of nitrogen gaseous diffusion at high temperature. Nitrogen enters into the material by diffusion, creating a surface layer of increased hardness. Low cycle fatigue behavior in air of Ti6Al4V alloy has been studied. Results show a reduction of low cycle fatigue life up to 10% compared to the not-treated material. Studies suggest it is not related to the titanium nitride surface layer, but to microstructural changes caused by the high temperature treatment. PMID- 15348344 TI - In vivo/ex vivo cellular interactions with titanium and copper. AB - Machined, commercially pure titanium (Ti) disks were coated with approximately 400 nm copper (Cu) by physical vapor deposition or left uncoated. The kinetics of inflammatory cell recruitment, distribution and viability was evaluated around Ti, Cu, and in sham sites after 1, 3, 12, 18, 24, and 48 h in a rat subcutaneous (s.c.) model. Further analysis of the cells on implant surfaces was performed by ex vivo incubation of the disks. Ti and Cu stimulated an increased recruitment of inflammatory cells in comparison with sham sites. A markedly higher amount of cells, predominantly polymorpho-nuclear granulocytes (PMN), was detected around Cu after 18 h and onwards. More cells were found at the implant surfaces than in the surrounding exudates after 18 h. The total amount of lactate dehydrogenase (LDH), an indicator of plasma membrane injury, was higher in Cu exudates after 18 h in comparison with Ti and sham. In contrast, no differences in the proportion of dead cells (trypan blue dye uptake) were detected in the exudates. Further, LDH levels were higher around Ti than Cu during the initial 18 h of ex vivo incubation. The results of this study indicate that the early inflammatory process associated with a cytotoxic material in soft tissues is largely attributed to the induction of a markedly strong and prolonged chemotactic response. In contrast, this process is characterized by a higher amount of inflammatory cells around a biocompatible material than in sham sites, but with a transient course and total LDH similar to sham sites. PMID- 15348345 TI - The interaction of the orthopaedic metals, chromium VI and nickel, with hepatocytes. AB - High levels of metal ions, such as chromium and nickel, released from metallic total hip implants have been detected in the serum and urine of patients. Cr VI and Ni ions are carcinogenic and toxic and there is concern about their systemic toxicity. To investigate this we have studied the interaction of Cr VI and Ni with hepatocytes. Both metal ions caused loss of cell viability within 3 h exposure, Cr VI was more potent than Ni. Cr VI caused depletion of intracellular reduced glutathione (GSH) levels, and inhibition of glutathione reductase and glutathione-S-transferase (GST) activities. Expression of alpha-GST, the major isoenzyme of GST in rat liver, was also decreased by Cr VI. Ni, on the other hand did not deplete GSH, or inhibit any of the enzyme activities measured in the cells. GSH and GST form a major protection and detoxification system in the liver, and depletion of GSH and inhibition of GST activity by Cr VI in vivo may severely compromise the ability of an individual to protect himself against carcinogenic and cytotoxic chemicals in the environment. PMID- 15348346 TI - Cytotoxic and apoptotic effects of cobalt and chromium ions on J774 macrophages - Implication of caspase-3 in the apoptotic pathway. AB - The aim of this study was to evaluate the cytotoxic and apoptotic effects of cobalt and chromium ions on macrophages in vitro, and analyze the implication of caspase-3 in the apoptotic pathway. J774 mouse macrophages (5 x 10(5) cells/ml) were exposed for up to 24 h to 0-10 ppm Co2+ and 0-500 ppm Cr3+. The cytotoxic effect of ions was measured by Trypan blue exclusion. DNA analysis on agarose gel was used as a specific test for detection of DNA fragmentation into oligonucleosomes that occurs in apoptotic cells. The proteolytic cleavage of poly(ADP-ribose)polymerase (PARP), closely associated with the induction of apoptosis, was also analyzed along with the appearance of the active fragment of caspase-3, implicated in several apoptosis pathways. Results demonstrated that both Co2+ and Cr3+ ions induce macrophage mortality in a dose-dependent manner. However, Co2+ is more toxic inducing a cell mortality up to 28% with only 10 ppm vs. 37% with 500 ppm of Cr3+. DNA analysis demonstrated that both Co2+ and Cr3+ ions induce DNA fragmentation, between 6-10 ppm Co2+ and 250-500 ppm Cr3+ after 24 h incubation. PARP cleavage and the appearance of caspase-3 active fragment were observed after 6 h with both Co+ and Cr3+ ions, with a stronger signal after 24 h and 10 ppm of Co2+ or 500 ppm of Cr3+. In conclusion, this study demonstrates that after 24 h incubation, both Co2+ and Cr3+ ions can induce macrophage mortality, and more specifically apoptosis. The results also suggest that apoptosis occurs via a caspase-3 pathway. However, the relative importance of necrosis and apoptosis and the effects of longer exposure times on the induction of macrophage death by these metal ions remain to be investigated. PMID- 15348347 TI - Induction of apoptosis in human microvascular endothelial cells by divalent cobalt ions. Evidence for integrin-mediated signaling via the cytoskeleton. AB - Wound healing following implantation is characterized by an acute inflammatory reaction and a subsequent reorganizing phase in which angiogenesis is involved. Endothelial cells (EC) participate in both inflammation and angiogenesis. Thus, the effects on functions of EC exerted by implanted materials could affect the progression of wound healing. The corrosion of metallic implants can cause high concentrations of heavy metal ions in the peri-implant tissues. The purpose of the present study was to test the effects of possible corrosion products on the function and viability of human EC in vitro. Long-term exposure of EC to CoCl2 and NiCl2 (3 days, 0.7 mM) leads to a decrease of cell number and changes in cellular morphology. However, the morphological changes between CoCl2- and NiCl2 treated cells differ significantly. The changed morphology of CoCl2-treated EC and the fragmented DNA pattern indicates apoptosis. Nickel-treated cells demonstrated necrosis. The activity of integrins was tested by an assay of cellular adhesion on collagen-coated surfaces. It was shown that the number of adherent cells significantly decreased upon exposure to CoCl2. Our studies suggest that induction of cell death in EC upon exposure to CoCl2 could be attributed to impaired integrin signaling, which leads to a damaged cytoskeleton and culminates in apoptosis. PMID- 15348348 TI - The influence of polymeric membrane surface free energy on cell metabolic functions. AB - In membrane bioartificial organs using isolated cells, polymeric semipermeable membranes are used as immunoselective barriers, means for cell oxygenation and also as substrata for adhesion of anchorage-dependent cells. The selection of cytocompatible membranes that promote in vitro cell adhesion and function could be dependent on its membrane properties. In this study we investigated the physicochemical aspects of the interaction between the membrane and mammalian cells in order to provide guidelines to the selection of cytocompatible membranes. We evaluated the metabolic behavior of isolated liver cells cultured on various polymeric membranes such as the ones modified by protein adsorption. The physico-chemical properties of the membranes were characterized by contact angle measurements. The surface free energy of membranes and their different parameters acid (gamma+), base (gamma-) and Lifshitz-van der Waals (gammaLW) were calculated according to Good-van Oss's model. The adsorption of protein modified markedly both contact angle and membrane surface tension. In particular, membrane surface free energy decreased drastically with increased water contact angle. For each investigated membrane we observed that liver specific functions of cells improve on hydrophilic membrane surfaces. For all investigated membranes the rate of ammonia elimination increased with increasing of membrane surface free energy. PMID- 15348349 TI - Adhesion strength of individual human bone marrow cells to fibronectin. Integrin beta1-mediated adhesion. AB - The purpose of this work was to study the adhesion strength of individual bone marrow cells, using a micropipette aspiration technique. The adhesion strength of the primary human bone marrow cells to fibronectin-coated substrate, by blocking the beta1 integrin with and without antibodies, was also determined. Human bone marrow stromal cells of the second passage were seeded at a density of 500 cells/cm2 on two different substrates: plastic culture dish (PCD) and PCD coated with fibronectin. In short adhesion times (15-180 min) the cells attached without spreading and remained almost spherical. A negative pressure of about 3500 Pa was applied, through the micropipette, on individual bone marrow cells and the detach process was recorded. The tip of the micropipette was bent at an 130 degrees angle to the corpus of the pipette and it was manipulated to be on the upper side of the cell and vertically to the bottom of the plate. It was observed from the experiments that the cells exhibited smaller adhesion strength at early adhesion times (30-85 min). After 85 min the adhesion strength increased abruptly and remained relatively constant for the adhesion period from 85 to 180 min for all substrates. Monoclonal antibodies against integrin subunit beta1 were used for integrin blocking experiments. The data suggested that the attachment of osteoblasts to a plastic culture dish without fibronectin coating occurred earlier than to the one coated with fibronectin PCD. In longer adhesion time the coating with fibronectin increased the adhesion strength at 107%. Blocking of integrin beta1 with monoclonal antibody resulted in decrease of the adhesion strength at 49%. PMID- 15348350 TI - Monocyte adhesion and adhesion molecule expression on human endothelial cells on plasma-treated PET and PTFE in vitro. AB - The aim of this study was to evaluate in vitro the inflammatory potential of endothelialized surfaces of polyethylene terephthalate (PET) and polytetrafluorethylene (PTFE) after ammonia gas plasma modification. HUVECs grown on polystyrene and HUVECs stimulated with tumor necrosis factor (TNF-alpha) were used as controls. At day 1 and day 7, surfaces were evaluated for U937 cells and HUVECs using flow cytometry and immunohistochemistry. Plasma-treated PET (T-PET) and treated PTFE (T-PTFE) increased U937 cell adhesion compared to the negative control but this was not statistically significant. Maximal adhesion of U937 cells to HUVEC was observed on TNF-alpha stimulated endothelium with significant differences between day 1 and day 7. There was a small increase in U937 cell adhesion to plasma-treated PET compared to PTFE on both day 1 and day 7, but this was not statistically significant. Immunohistochemical staining demonstrated two patterns of distribution for monocyte adhesion on materials. On T-PET the cells were positioned in clusters attached to HUVECs and on T-PTFE the cells were randomly distributed on HUVECs and material. The effects of plasma-treated PET and PTFE on HUVEC adhesion and proliferation were also studied. On day 1 there were slight increases in the growth of HUVECs on both of T-PET and T-PTFE but this was not statistically significant. On day 7, cell number increased significantly on all of surfaces compared to the negative control. The results demonstrate that the plasma treatment of PET and PTFE with ammonia improves the adhesion and growth of endothelial cells and these surfaces do not exhibit a direct inflammatory effect in terms of monocyte adhesion. Plasma-treated PTFE enhances HUVECs growth and was less adhesive for monocytes as compared with treated PET. The monocyte adhesion to endothelial cells on surfaces can be used as a tool for the evaluation of material surface modification and further to study the mechanisms of cell to cell interactions in response to surfaces. PMID- 15348351 TI - Binding of vitronectin and clusterin by coagulase-negative staphylococci interfering with complement function. AB - Coagulase-negative staphylococci (CoNS) are commonly associated with infections of prosthetic devices mediated by adsorbed host factors on biomaterial surfaces. Complement activation is known to occur and induce unspecific inflammation around the biomaterials. Human vitronectin (Vn) and clusterin (Clu), two potent inhibitors of complement, can be bound by CoNS. With a hypothesis whether binding of Vn or Clu influences complement activation, two measurements were determined. For Vn, complement activation was measured with a mouse anti-activated human C9 antibody. In the presence of Vn-binding strain, Staphylococcus hemolyticus SM13I, complement activation on a surface pre-coated with Vn occurred as it did in the absence of Vn pre-coating. For S. epidermidis 3380, which does not express binding of Vn, complement activation on a Vn-presented surface was significantly decreased. For Clu, erythrocytes lysis was measured to reflect the end product of complement activation (membrane attack complex). The complement-induced hemolysis increased when human serum was pre-incubated with Clu-binding strains, S. epidermidis J9P. The enhancement of hemolysis by J9P decreased when serum was supplemented by exogenous Clu. The data imply that interaction between CoNS and Vn or Clu interferes with one of their physiological functions, complement inhibition. PMID- 15348352 TI - Formulation of PEG-based hydrogels affects tissue-engineered cartilage construct characteristics. AB - The limited supply of cartilage tissue with appropriate sizes and shapes needed for reconstruction and repair has stimulated research in the area of hydrogels as scaffolds for cartilage tissue engineering. In this study we demonstrate that poly(ethylene glycol) (PEG)-based semi-interpenetrating (sIPN) network hydrogels, made with a crosslinkable poly(ethylene glycol)-dimethacrylate (PEGDM) component and a non-crosslinkable interpenetration poly(ethylene oxide) (PEO) component, and seeded with chondrocytes support cartilage construct growth having nominal thicknesses of 6 mm and relatively uniform safranin-O stained matrix when cultured statically, unlike constructs grown with prefabricated macroporous scaffolds. Even though changing the molecular weight of the PEO from 100 to 20 kDa reduces the viscosity of the precursor polymer solution, we have demonstrated that it does not appear to affect the histological or biochemical characteristics of cartilaginous constructs. Extracellular matrix (ECM) accumulation and the spatial uniformity of the ECM deposited by the embedded chondrocytes decreased, and hydrogel compressive properties increased, as the ratio of the PEGDM:PEO in the hydrogel formulation increased (from 30:70 to 100:0 PEGDM:PEO). Total collagen and glycosaminoglycan contents per dry weight were highest using the 30:70 PEGDM:PEO formulation (24.4+/-3.5% and 7.1+/-0.9%, respectively). The highest equilibrium compressive modulus was obtained using the 100:0 PEGDM:PEO formulation (0.32+/-0.07 MPa), which is similar to the compressive modulus of native articular cartilage. These results suggest that the versatility of PEG based sIPN hydrogels makes them an attractive scaffold for tissue engineering of cartilage. PMID- 15348353 TI - Urothelial cell transplantation using biodegradable synthetic scaffolds. AB - Reconstruction of the urinary bladder with bowel to restore storage capacity is associated with significant complications arising from substituting an absorptive, mucus-producing intestinal epithelium for the barrier urothelium of the bladder. To overcome these problems, we are developing a "composite enterocystoplasty" procedure to replace the epithelium of the bowel with autologous in vitro-propagated normal urothelial cells. The aims of this study were to evaluate synthetic biomaterials as delivery vehicles for the cultured urothelial cells and provide support during transfer and cell adherence to the de epithelialized bowel wall. The surgical compliance of 12 biomaterials was evaluated, along with their ability to support urothelial cell attachment. Transfer of urothelial cells onto biomaterials as single cell suspensions or intact cell sheets was investigated. Seeding of a single cell suspension on to non-woven mesh resulted in poor cell attachment. Seeding onto woven mesh was more efficient, but the most effective transfer method involved producing an intact cell sheet that could be combined with woven, knitted and non-woven biomaterials. Transfer of the cell sheet : mesh complexes onto a de-epithelialized bladder stroma produced a stratified epithelium incorporating the strands of the mesh and expressing urothelial-associated antigens after 48 h in organ culture. Thus, we have developed and evaluated a suitable transfer method for in vitro propagated urothelial cells to be used in "composite enterocystoplasty". PMID- 15348354 TI - Osteoblast viability and detachment following exposure to ultrasound in vitro. AB - Ultrasound has been used in dentistry for over 40 years and has recently been proposed for cutting bone. The purpose of this study was to establish the effects of ultrasonic instruments on osteoblasts. A 25 kHz magnetostrictive ultrasound generator and a TFI-1 tip (Dentsply, UK) were used as the ultrasound generating instruments. Primary osteoblast cultures were established from the parietal bones of two-day-old Albino Wistar rats grown on tissue culture (TC) petri dishes (Corning, UK) in alphaMEM (Sigma, UK). Once confluent, the osteoblasts were harvested using 0.05% trypsin in 0.02% EDTA then 1.7 x 10(5) cells in 2.5 ml of alphaMEM were either re-seeded immediately onto TC dishes and allowed to adhere for 24 h or kept in suspension before application of ultrasound with different tip displacements prior to re-seeding the cells. Osteoblast viability was assessed using 0.4% Trypan Blue following the initial dose of ultrasound then periodically over a 20 h period for both adherent and suspension osteoblasts. This study demonstrated that ultrasound caused osteoblast detachment and loss of viability in vitro, both when adherent to a substrate or in suspension. Loss of osteoblast viability was related to the maximum displacement of the ultrasonic tip and continued throughout the 20 h period observed for osteoblasts adherent to TC dishes. PMID- 15348355 TI - Effects of therapeutic ultrasound on osteoblast gene expression. AB - Ultrasound (US) is commonly used as a physiotherapy aid for a number of types of injury to soft connective tissues and for fracture healing. However, the precise effects of therapeutic US on tissue healing processes are not clearly understood, although they are likely to involve changes in key cellular functions. The present study has therefore examined the effects of several US intensity levels on the activity of two bone-associated proteins, alkaline phosphatase (ALP) and osteopontin (OP) in a human cell line, MG63, using RT-PCR. ALP showed progressively higher expression with increasing US intensities, whereas OP responded differently, showing down-regulation at 120 mW/cm2, the lowest US exposure. OP expression was considerably less affected overall compared with the relative response of ALP to the same US doses. The results show that there is a differential response to therapeutic levels of US, since ALP and OP clearly exhibited gene-specific response profiles. These findings suggest that modifying the parameters of US exposure could be used to improve repair and regeneration processes and enhance the clinical efficacy of implanted biomaterials for tissue engineering. PMID- 15348356 TI - A study of the cellular response to orientated fibronectin material in healing extensor rat tendon. AB - 3D orientated fibronectin (Fn) mats have been used as biocompatible and biodegradeable scaffolds to provide orientated cues using contact guidance for cell migration/adhesion and deposition of extracellular matrix. We have implanted Fn scaffolds in an established rat tendon(partial tenotomy) injury model to test its efficacy and monitor the early cellular and inflammatory response. Tendons were harvested at 0, 6 h, 1, 3, 5, 7 and 14 days for H&E, immunohistochemistry and TEM. Total cell counts within the window increased progressively with time with no significant differences between the Fn scaffolds and controls. CD45 (pan leukocyte) positive cell numbers peaked at 6 h and when expressed as a percentage of total cell counts as determined by H&E staining constituted 20% of the total cell number at 6 h but decreased to 5% of total number by 72 h. There were no significant differences in the inflammatory response between the control and implanted groups. Few CD44 (mesenchymal stem cell) positive cells identified had a surface location. A novel cell with long exaggerated cytoplasmic processes was identified by TEM. Our results show that the Fn scaffold did not degrade or elicit any untoward inflammatory response at the time points tested and has potential use in guiding the repair process. PMID- 15348357 TI - Calcification and identification of metalloproteinases in bovine pericardium after subcutaneous implantation in rats. AB - The purpose of this study was to evaluate the influence of two anticalcification pre-treatments (chloroform/methanol and ethanol) and serum conditioning of glutaraldehyde-crosslinked bovine pericardium on the calcification degree and the presence of gelatinase activities in a subcutaneous implantation model in rats. Regarding calcification of the implants, glutaraldehyde control treatments showed a significatively higher calcification degree than pericardium treated with anticalcification reagents. Serum conditioning of glutaraldehyde treated tissues did not influence the calcification degree; moreover, no differences were found in these samples with the time of implantation (30 and 90 days). On the other hand, anticalcification treatments resulted in a very significant decrease in the calcium content in the implanted membranes. Gelatinase activities were detected by gelatin zymography in almost all the implanted samples. However, control tissues with and without serum conditioning showed less gelatinase activities than those samples pre-treated with anticalcification treatments. Metalloproteinase (MMP-2) activity was detected in all the samples analyzed but a higher expression of MMP-9 was detected in those implants treated with chloroform/methanol and ethanol. Additional gelatinase activities showing lower molecular weight than MMP-2 were also detected in both anticalcification treated samples. The presence of these gelatinase activities is probably due to host cellular infiltrates and could contribute to the biomaterial degradation. PMID- 15348358 TI - On the formation of fibrous capsule and fluid space around machined and porous blood plasma clot coated titanium. AB - Machined and machined submicron porous titanium, with and without a thin blood plasma coating (100 nm), were implanted for 7 or 28 days in subcutaneous pockets on the back of the rat. After explantation the specimens were analyzed by light microscopy with respect to thickness of the fibrous capsule, the fluid space width between implants and fibrous capsule, and formation of blood vessels. The results at 7 days indicate a thinnest fluid space for the plasma clot coated porous titanium surface, and the spaces vanished at the light microscopic level after 28 days outside all the analyzed surfaces. The thickness of the fibrous capsule increased outside the different surfaces at 7-28 days, and in this respect no significant differences were observed between the different surfaces at any time. Analysis of neovascularization showed that the number of vessels and proportion of vessels in the fibrous capsule increased with time at all surfaces, except machined Ti where the number instead decreased from 7 to 28 days. The average distance between the blood vessels and the fluid space increased with time for all types of surfaces. The results in the present study indicate that the healing process around titanium can be modulated by porosity and thin pre prepared plasma coatings. PMID- 15348359 TI - Oxidized implants and their influence on the bone response. AB - Surface oxide properties are regarded to be of great importance in establishing successful osseointegration of titanium implants. Despite a large number of theoretical questions on the precise role of oxide properties of titanium implants, current knowledge obtained from in vivo studies is lacking. The present study is designed to address two aspects. The first is to verify whether oxide properties of titanium implants indeed influence the in vivo bone tissue responses. The second, is to investigate what oxide properties underline such bone tissue responses. For these purposes, screw-shaped/turned implants have been prepared by electrochemical oxidation methods, resulting in a wide range of oxide properties in terms of: (i) oxide thickness ranging from 200 to 1000 nm, (ii) the surface morphology of barrier and porous oxide film structures, (iii) micro pore configuration - pore sizes<8 microm by length, about 1.27 microm2 to 2.1 microm2 by area and porosity of about 12.7-24.4%, (iv) the crystal structures of amorphous, anatase and mixtures of anatase and rutile type, (v) the chemical compositions of TiO2 and finally, (vi) surface roughness of 0.96-1.03 microm (Sa). These implant oxide properties were divided into test implant samples of Group II, III, IV and V. Control samples (Group I) were turned commercially pure titanium implants. Quantitative bone tissue responses were evaluated biomechanically by resonance frequency analysis (RFA) and removal torque (RT) test. Quantitative histomorphometric analyses and qualitative enzyme histochemical detection of alkaline (ALP) and acidic phosphatase (ACP) activities were investigated on cut and ground sections after six weeks of implant insertion in rabbit tibia. In essence, from the biomechanical and quantitative histomorphometric measurements we concluded that oxide properties of titanium implants, i.e. the oxide thickness, the microporous structure, and the crystallinity significantly influence the bone tissue response. At this stage, however, it is not clear whether oxide properties influence the bone tissue response separately or synergistically. PMID- 15348360 TI - A new titanium fiber mesh-cuffed peritoneal dialysis catheter: an experimental animal study. AB - CAPD catheters are associated with infectious complications. To solve this problem, we developed a new catheter. In our design, sintered titanium fiber mesh material replaced the Dacron cuffs, as used in standard Tenckhoff catheters. The purpose of the current study was to compare the tissue response to new titanium cuffed vs. Dacron-cuffed catheters. Experimental and standard Tenckhoff catheters were inserted intraperiotoneally in 12 goats, using a so-called two-stage surgical technique. In the first surgical session, the catheters were implanted. However, the percutaneous part of the catheter was buried subcutaneous. After 3-5 weeks, the percutaneous part of the catheter was exteriorized. After 14 weeks of implantation, all implants with surrounding tissue were retrieved and prepared for histological evaluation. Subsequently, we quantified: the characteristics of the fibrous tissue capsule surrounding the cuffs, the tissue inside the cuff porosity, and the epidermal downgrowth. Histologic and histomorphometric evaluation showed that titanium mesh evoked a lesser inflammatory response inside the cuff porosity compared with Dacron cuffs. Besides, the fibrous tissue capsule surrounding the titanium cuffs was significantly thinner. Supported by the obtained results, we conclude that the use of titanium fiber mesh has a great potential for application in percutaneous devices. PMID- 15348361 TI - Comparison of wear in a total knee replacement under different kinematic conditions. AB - A six station ProSim (Manchester, UK) knee simulator was used to assess the wear of six PFC (DePuy) fixed bearing total knee replacements under two different kinematic conditions defined as low and high kinematic inputs. The high kinematics displacement and rotation inputs were based on the kinematics of the natural knee with ISO standards used for the axial load and flexion. Low kinematics were defined as approximately half the magnitude. The six specimens were run for three million cycles under low kinematics and three million cycles under high kinematics. The mean wear rate found during the low kinematics phase was 7.7 +/- 2 mm3 per million cycles. This then increased significantly to an average wear rate of 41 +/- 14 mm3 during the high kinematics input phase. The wear areas were characterized by a predominant damage mode of burnishing with some abrasive wear occurring during the high kinematics phase. This study supports the findings that introduction of cross-shearing of the polyethylene by introducing both rotational and anterior/posterior displacement increases the wear rate. This has implications for younger patients with higher levels of activity that need knee replacements. PMID- 15348362 TI - The effect of accelerated aging on the wear of UHMWPE. AB - Oxidative degradation of UHMWPE has been found to be a cause of elevated wear rate of the polymer in total joint replacement leading to failure of these devices. In order to evaluate long term stability of polymers, various accelerated aging methods have been developed. In this study, wear rates of shelf aged UHMWPE and "accelerated aged" UHMWPE were compared using a multi-directional pin-on-plate wear test machine in order to evaluate the effect of the accelerated aging on wear. Wear factors of the aged materials were found to depend on their density, which is a measure of oxidation level. Finally, accelerated aging was calibrated against shelf aging in terms of wear rate. PMID- 15348363 TI - Wear of fixed bearing and rotating platform mobile bearing knees subjected to high levels of internal and external tibial rotation. AB - In order to extend the lifetime of total knee replacements (TKR) in vivo, reduction of the volumetric wear rate of ultra high molecular weight polyethylene (UHMWPE) bearings remains an important goal. The volume of wear debris generated in fixed bearing total knee devices increases significantly when subjected to higher levels of internal-external rotation and anterior-posterior displacement. Six PFC Sigma fixed bearing TKR were compared with six LCS rotating platform mobile bearing knees using a physiological knee simulator with high rotation kinematic inputs. The rotating platform polyethylene inserts exhibited a mean wear rate which was one-third of that of the fixed bearing inserts despite having increased femoral contact areas and additional tibial wear surfaces. The rotating platform design decouples knee motions, by allowing unidirectional motion at the tray-insert articulation, which reduces rotation at the femoral-insert counterface. This translation of complex knee motions into more unidirectional motions results in molecular orientation of the UHMWPE and reduced volumetric wear. PMID- 15348364 TI - Long-term wear of HIPed alumina on alumina bearings for THR under microseparation conditions. AB - The long term wear and wear debris generated in HIPed alumina on alumina bearings for hip prostheses with microseparation in vitro is compared to standard simulator conditions and ex vivo specimens. Microseparation studies were completed to five million cycles at two severity levels in attempts to rigorously evaluate the long-term tribological performance of the bearings. During the first million cycles (bedding-in) of the microseparation tests characteristic stripe wear was observed on all of the femoral heads with a matching area on the rim of the acetabular inserts. Under mild microseparation conditions an average wear rate of 0.55 mm3/million cycles was observed during the initial million cycles which reduced to a steady state level of 0.1 mm3/million cycles. Under more severe conditions an average wear rate of 4.0 mm3/million cycles was observed during bedding-in which reduced to a steady state level of 1.3 mm3/million cycles. These compare to a bedding-in wear rate of 0.11 mm3/million cycles and steady-state wear rate of 0.05 mm3/million cycles for the same material under normal simulation with no microseparation. Furthermore, under microseparation the wear mechanisms and wear debris were similar to those observed in previous alumina retrieval studies with debris ranging from 10 nm to 1 microm in size. PMID- 15348365 TI - Implications of orthopedic fretting corrosion particles on skeletal muscle microcirculation. AB - Particulate corrosion and wear products of metal implants are increasingly becoming topics of interest, due to the cascade of biological and biomechanical events they induce. The impairment of skeletal muscle microcirculation by fretting corrosion particles may have profound consequences. We therefore studied in vivo leukocyte-endothelial cell interaction in skeletal muscle after confrontation with characterized titanium and stainless steel fretting corrosion particles, and compared these results with those of the bulk materials. Using the hamster dorsal skinfold chamber preparation and intravital microscopy, we could demonstrate that stainless steel induces a more pronounced inflammatory answer in contrast to the implant material titanium. However, we were not able to show a general benefit of bulk vs. debris. Overall, the study suggests that not only the bulk properties of orthopaedic implants, but also the microcirculatory implications of inevitable wear debris, may play a role in determining biocompatibility and ultimately longevity of an implant. The skinfold chamber is a feasible and versatile model for observation of the dynamic process of microvascular response after foreign-body implantation, and offers much perspective. PMID- 15348366 TI - Phenotypic characteristics of T lymphocytes in the interfacial tissue of aseptically loosened prosthetic joints. AB - Aseptic loosening of prosthetic joints represents a major cause for revision surgery. Wear particles represent the initial stimulus in the production of a multi-cellular inflammatory membrane at the bone-implant interface. The mechanisms by which this membrane is established and its influence on bone resorption are central to understanding aseptic loosening. T cells have been identified as a component of interface tissue a feature confirmed in this study. Of the 15 cases studied, 12 cases (12/15) stained positive for CD3. T cell infiltration was present throughout the sections with some perivascular clustering. Multiplex PCR (MPCR) testing of eight of the 15 cases for Th1/Th2 cytokines did not show a predominance of either "type" of T cell response. Interleukin (IL)-2 mRNA expression was the most common feature (7/8) while IL-4 (6/8), IL-13 (6/8) and IFN-gamma mRNA expression (6/8) was also prevalent. IL-5 (4/8) and IL-10 (4/8) mRNA expression was less and IL-12 (3/8) mRNA expression was the least. Recent work has shown furthermore, that activated T cells can directly stimulate osteoclastogenesis through the expression of RANK ligand. However, although staining for RANK ligand was a consistent feature of all cases stained, such RANK ligand expression was limited to endothelial cells. Helper T cells control and develop immune responses, their role in the inflammation seen in aseptic loosening will aid further understanding of this reaction and may also identify key points for therapeutic intervention. PMID- 15348367 TI - Does a pro-angiogenic state exist in the bone-implant interface of aseptically loosened joint prosthesis? AB - Neovascularization is indispensable to both bone remodeling and the development of chronic inflammation. A pro-angiogenic state in the periprosthetic tissue may augment the inflammatory response to wear debris. To investigate if a pro angiogenic state exists in the bone-implant interface of aseptically loosened joint prosthesis, the expression of vascular endothelial growth factor (VEGF) and its receptor Flk-1/KDR were studied by immunohistochemistry. The VEGF-Flk/KDR pathway has been implicated as a key signaling requirement for pathological angiogenesis. The level of vascularization in periprosthetic tissue was semi quantitatively compared to osteoarthritic (OA) and rheumatoid arthritic (RA) synovium. The level of vascularization in areas of periprosthetic tissue with heavy or low/moderate wear debris were also compared semi-quantitatively by image analysis. High levels of VEGF expression (16/16 cases) particularly in the implant synovial-like lining layer together with Flk-1/KDR expression by endothelial cells (13/16), suggests that neovascularization is occurring. Morphometric comparison of periprosthetic tissue with RA and OA synovium revealed no significant difference in microvessel density, but did reveal significantly increased microvessel area in RA synovium (P > 0.05). Areas of high wear debris infiltrate also contained a significantly smaller microvessel area (P > 0.01). Suggesting that wear debris may cause behavioral modification of microvessels. Modifying angiogenesis in the periprosthetic tissue could be a potential therapeutic target in reducing inflammation. PMID- 15348368 TI - Functionalized surfaces for optical biosensors: applications to in vitro pesticide residual analysis. AB - Functionalized biosensing surfaces were developed for chemiluminescent immunoassay of pesticides. Two approaches to construct functionalized surfaces were tested: (i) pesticide is immobilized to the surface and interacts with a labeled antibody; (ii) antibody is immobilized and interacts with a labeled pesticide. As labels alkaline phosphatase and peroxidase were used with their corresponding substrates CSPD and luminol, respectively. Light produced by chemiluminescent substrate was detected by a thermoelectrically cooled CCD camera or a photomultiplier. The best detection limit 0.00001 ng/ml was obtained using antibodies immobilized to dextran-enhanced surface. Completely renewable surface was obtained using reversible lectin-monosaccharide interaction, one surface was used for 200 analyses without any loss of binding capacity. Most favorable stability and cost per analysis was achieved with molecularly imprinted polymer (MIP) instead of antibody. The functionalized biosensing surfaces were prepared to detect 2,4-dichlorophenoxyacetic (2,4-D) acid as a model pesticide. The developed concepts are, however, generally applicable to other pesticides and to other optical formats, e.g. optical fiber. PMID- 15348369 TI - Glucose biosensors based on dendrimer monolayers. AB - The peculiarities of glucose biosensors based on different generation of dendrimers (G0, G1 and G4) have been studied by amperometry and QCM techniques. It is shown that stable glucose biosensor can be obtained with low generation of dendrimers. The sensor sensitivity, however considerable, increased with increasing number of generation of dendrimers. This can be due to the increased volume of the dendrimer interior as well as with increased number of binding sites for glucose oxidase (GOX). QCM experiments showed that immobilization of GOX resulted in formation of enzyme multilayers on a dendrimer surface. The enzyme turnover for this system (0.1-0.01 s(-1)) was lower then that for immobilization of GOX onto a supported lipid films by means of avidin-biotin technology (1.1 s(-1)). However, dendrimer based biosensors are more stable in comparison with sBLM based sensors and could be stored in a refrigerator in dry conditions over 15 days without substantial loss of sensitivity. PMID- 15348370 TI - Effect of water soluble extract of nacre (Pinctada maxima) on alkaline phosphatase activity and Bcl-2 expression in primary cultured osteoblasts from neonatal rat calvaria. AB - The nacre (mother of pearl) layer of the oyster Pinctada maxima shell can initiate bone formation by human osteoblasts in vivo and in vitro and is a new biomaterial that induces osteogenesis. This activity of nacre could be due to its water-soluble matrix. We examined the action of a water-soluble extract of nacre on the osteoblast phenotype of cells isolated from rat neonatal calvaria by measuring alkaline phosphatase (ALP) activity and by localization of the anti apoptotic protein Bcl-2 by immunocytochemistry. ALP activity was increased 7% (p<0.001) by 100 microg proteins/ml extract and 20% (p<0.001) by 50 microg proteins/ml extract, but a low concentration of extract decreased the ALP activity by 8%. Cells treated with a high aspartic acid content fraction of the extract had increased ALP activity (23%, p<0.0001). Nacre extract and the fraction have no effect on the proliferation of mature osteoblasts. Immunoreactive Bcl-2 was overproduced in the cytoplasm and nuclei of osteoblasts at all stages of culture. Bcl-2 was found over the whole chromatin in quiescent and mitotic cells at the end of mitosis in the two nuclei in one cell, before cytodieresis. Bcl-2 was also found over chromosomes. Thus, nacre extract stimulates Bcl-2 production in osteoblasts, that is correlated with the cell cycle. Bcl-2 was also abundant in the nucleoli of extract-treated cells. Thus, the concentration and subcellular distribution of Bcl-2 in osteoblasts in primary cultures is influenced by nacre extract, and related to the cell cycle and the regulation of gene expression. Hence, knowledge of how water-soluble extracts of Pinctada maxima nacre act on osteoblasts in vitro may reveal the mechanisms involved in its action in vivo on bone cells and bone regeneration. PMID- 15348371 TI - Bone formation induced by calcium phosphate ceramics in soft tissue of dogs: a comparative study between porous alpha-TCP and beta-TCP. AB - Two kinds of tri-calcium phosphate ceramics (Ca/P = 1.50), alpha-TCP and beta TCP, which has the same macrostructure and microstructure, but different phase composition, were implanted in dorsal muscles of dogs. The samples were retrieved at 30, 45 and 150 days, respectively, after implantation, and were analyzed histologically. There were critically different tissue responses between alpha TCP ceramic and beta-TCP ceramic. Higher cell populations were observed inside the pores of beta-TCP than those of alpha-TCP, bone tissue was found in beta-TCP at 45 and 150 days, but no bone formation could be detected in any alpha-TCP implants in this study. On the other hand, the bone tissue in beta-TCP seemed to degenerate at 150 days. The results indicate that porous beta-TCP can induce bone formation in soft tissues of dogs; while the rapid dissolution of the ceramic and the higher local Ca2+, PO(4)3- concentration due to the rapid dissolution of alpha-TCP may resist bone formation in alpha-TCP and the less rapid dissolution of beta-TCP may be detrimental to already formed bone in beta-TCP. PMID- 15348372 TI - Design of a simulated urethra model for the quantitative assessment of urinary catheter lubricity. AB - Catheters designed for intermittent urological catheterization should possess appropriate lubricity and mechanical properties to ensure optimal clinical performance. However, the lack of a reproducible in vitro method that simulates clinical use makes it difficult to evaluate the lubricity of urinary catheters and other devices for urethral insertion. Therefore, this study describes a suitable method based on use of a Texture Analyzer to characterize the lubricity of such devices. The novel method was susequently applied to the evaluation of commercially-available intermittent urinary catheters. In addition, other important physicochemical properties of these catheters were examined, namely Young's modulus, degree of hydration and morphology. Catheter lubricity was quantified, using a Texture Analyzer, by measurement of the forces required for insertion and removal of the device from two model substrates, agar and mucin coated silicone tubing. Significant differences in lubricity were identified between the commercially-available catheters, with Aquacath and Lofric exhibiting the lowest forces of insertion and removal. There were no significant differences between the extent of hydration between the catheters, with the exception of Uro flo which exhibited the lowest hydration. Therefore, the differences in lubricity were not directly related to the extent of hydration. The forces required for insertion/removal of all catheters were markedly greater in the simulated mucin model than in the agar substrate and the former, simulated urethra model, was accepted to mimic more accurately, the in vivo situation. Significant differences were observed between the Young's Moduli of the catheter biomaterials, with Aquacath possessing the largest value. In conclusion, this study has described the use of a texture analyzer and polymeric substrates for the evaluation of biomaterial lubricity. Using these methods, Aquacath and Lofric catheters exhibited greatest lubricity. However, following additional consideration of the mechanical properties of these biomaterials, Aquacath possessed the most appropriate physicochemical properties for use in intermittent catheterization. PMID- 15348373 TI - A combined XPS-SEM/EDX investigation on explanted UHMW polyethylene acetabular cups: possible role of silicon traces in the wear debris. AB - An investigation was started aimed at a better understanding of the complex phenomena leading to chemical degradation and morphological deterioration of UHMW polyethylene cups in total hip prostheses. Analysis was performed on retrieved implants which needed revision due to inflammation and pain problems. Preliminary results obtained by parallel XPS and SEM/EDX experiments gave evidence, for the first time, that silicon traces are involved in the process of particle formation and segregation onto the surface of the cups. The extent of modification of the surface chemical composition of cups and the process of particle segregation seem to be correlated to both the implant time and to some particular features of patient (age, activity, style of life, etc.). Investigation on a large number of samples is in progress in order to test this hypothesis. The results obtained so far confirmed the potential of surface spectroscopies (XPS) in biomaterial investigations. PMID- 15348374 TI - The influence of gamma irradiation and aging on degradation mechanisms of ultra high molecular weight polyethylene. AB - The aging behavior of ultra-high molecular weight polyethylene (UHMWPE) has been studied following gamma irradiation in air. Accelerated aging procedures used elevated temperature (70 degrees C), pressurized oxygen (5 bar) and applied stress. Shelf and in vivo aged components have also been investigated. The variation in polymer properties with depth into the polymer was determined using density measurements, infra-red spectroscopy and differential scanning calorimetry. Accelerated aging in pressurized oxygen resulted in peaks in polymer density and degree of oxidation up to 500 microm below the polymer surface. Shelf and in vivo aging was also found to result in increased density at or below the component surfaces. Changes in density were mainly due to changes in crystallinity within the UHMWPE and, to a smaller extent, due to oxygen incorporation within the polymer. The application of stress did not appear to influence the accelerated aging of UHMWPE. A method for estimating the residual stress distribution in the UHMWPE using the measured changes in density is proposed. This study has indicated that oxidation of UHMWPE may lead to the development of tensile residual stresses, near the component surface, in the region of 1.7 MPa. These stresses may contribute to the failure mechanism of UHMWPE acetabular cups or knee tibial trays during service. PMID- 15348375 TI - Influence of loading types on the shear strength of the dentin-resin interface bonding. AB - Small differences in the shear bond test can make critical differences in the bonding strength values. The purpose of this study was to compare the influence of the orthodontic-looped wire, stainless steel tape and chisel systems used in shear bonding tests to verify the resistance in the dentin-resin interface. Forty eight human teeth were used and divided in three groups. The teeth were ground until a flat smooth surface was achieved, that was delimited with an adhesive tape containing a hole of 4 mm diameter. After, the dentine surface was treated with Scotchbond Multi Purpose and the composite Z-100 was applied in layers, through a stainless steel mold. The samples were stored at 37 degrees C and 100% of relative humidity for 24 h and, then, submitted to 500 thermal cycles. After, they were taken to an universal test machine (Otto Wolpert) with crosshead speed of 6 mm/min. The results were statistically analyzed using a Tukey's test (p?lt 0.05). The orthodontic-looped wire determined the highest values of shear bond (13.33 MPa), following by chisel (7.81 MPa) and stainless steel tape (4.87 MPa). The debonding values depend on a complex stress combination produced during the loading of the samples. Small variations in test methodologies give statistically different values for shear bond strength. Different shear strength methods in vitro make the comparative clinical performance of the resin filling materials difficult. PMID- 15348376 TI - Effect of mechanical grinding of MCPM and CaO mixtures on their composition and on the mechanical properties of the resulting self-setting hydraulic calcium phosphate cements. AB - Calcium bis-dihydrogenophosphate monohydrate (or monocalcium phosphate monohydrate, MCPM) is often used as the acid calcium phosphate in hydraulic calcium phosphate cement formulations. But commercial MCPM is not pure; it contains a small amount of orthophosphoric acid and moisture. Consequently, MCPM is difficult to mill and the powder is sticky and presents aggregates. Because granularity influences the mechanical properties of the hardened cements, a possible way to get around this difficulty that has been proposed is to premix it with other materials before grinding. We therefore ground commercial MCPM with CaO. A rapid decrease in the amount of MCPM was observed during mechanical grinding by a solid-solid reaction with calcium oxide. The final products were anhydrous or dihydrate dicalcium phosphate and/or hydroxyapatite or calcium deficient hydroxyapatite depending on the initial calcium-to-phosphate (Ca/P) ratio. The mechanical properties (compressive strength and setting time) of cements made from MCPM and CaO were affected whatever the Ca/P ratio as a consequence of the change in composition of the starting materials. Storage at different temperatures of MCPM and CaO mixtures manually ground in a mortar for only 2 min and without mechanical grinding did not affect their composition, but a decrease was observed in the compressive strength of cements made from these mixtures. PMID- 15348377 TI - Encapsulation of deoxyribonucleic acid molecules in silica and hybrid organic silica gels. AB - The encapsulation of small DNA molecules was attempted in pure silica and in hybrid polyvinyl alcohol-silica gels. The materials which were obtained were examined by nitrogen adsorption, and by 29Si and 31P NMR spectroscopy. The extraction of the DNA molecules from the gels was examined in a buffer aqueous solution as well as in an acidic medium. The results suggested that the DNA molecules remained trapped inside the gels due to a permanent bonding to the gel network. PMID- 15348378 TI - Effects of fiber length and volume fraction on the reinforcement of calcium phosphate cement. AB - A self-setting calcium phosphate cement (CPC) transforms into solid hydroxyapatite during setting at body temperature, and has been used in a number of medical and dental procedures. However, the inferior mechanical properties of CPC prohibits its use in unsupported defects, stress-bearing locations or reconstruction of thin bones. The aim of the present study was to strengthen CPC with fiber reinforcement, to examine the effect of fiber length and volume fraction, and to investigate the reinforcement mechanisms. Previous studies employed either short fibers for random distributions, or continuous fibers that were as long as the specimen size with preferred orientations such as unidirectional alignment. In the present study, a novel methodology was developed in which fibers several times longer than the specimen mold size were randomly mixed with the CPC paste to approximate the isotropy associated with short fibers, and at the same time achieve the high reinforcement efficacy associated with continuous fibers. Carbon fibers of 8 microm diameter were used with fiber lengths ranging from 3 mm to 200 mm, and fiber volume fraction from 1.9% to 9.5%. A three-point flexural test was used to fracture the specimens. Scanning electron microscopy was used to examine crack-fiber interactions and specimen fracture surfaces. The composite containing fibers of 75 mm in length at a volume fraction of 5.7% achieved a flexural strength about 4 times, and work-of-fracture 100 times, greater than the unreinforced CPC. It is concluded that randomly mixing the CPC paste with carbon fibers that were several times longer than the specimen mold size resulted in substantial improvements in strength and fracture resistance; the reinforcement mechanisms were crack bridging and fiber pullout; and fiber length and volume fraction were key microstructural parameters that determined the cement properties. PMID- 15348379 TI - Studies on gelatin-based sponges. Part III: a comparative study of cross-linked gelatin/alginate, gelatin/hyaluronate and chitosan/hyaluronate sponges and their application as a wound dressing in full-thickness skin defect of rat. AB - Novel cross-linked sponges composed of gelatin/alginate and gelatin/hyaluronate and chitosan/hyaluronate (GH, GA and CH, respectively) were prepared and compared. Six different sponges with or without silver sulfadiazine (AgSD) were applied on the full-thickness dorsal skin defect of Wistar rat. The histology and epidermal wound healing rates of the skin defects were investigated by light microscopy and computerized morphometry 5 and 12 days post-operatively. In our full-thickness wound model (diameter 1 cm), the AgSD-impregnated sponges showed good wound healing performances on the whole. However, there appeared meaningful differences of wound healing between the gelatin-based sponges (GH, GA) and the CH. GH with AgSD was found to show the best wound healing properties as a wound dressing resulting from histological findings and computerized morphometric analysis of epidermal healing. PMID- 15348380 TI - Properties of polyvinyl pyrrolidone/beta-chitosan hydrogel membranes and their biocompatibility evaluation by haemorheological method. AB - The semi-IPN hydrogel membranes of polyvinyl pyrrolidone (PVP) and beta-chitosan were synthesized by crosslinking beta-chitosan with glutaraldehyde. Hydrogel membranes were characterized by spectroscopic, swelling, thermal and mechanical properties. The in vitro biocompatibility of hydrogel membranes was studied by haemorheological method. These hydrogels have water contents in the range of 60 70% with a high fraction contributed by free water (> 45%). The gel composition, amount of cross-linking agent and swelling temperature plays an important role in swelling kinetics of these semi-IPN membranes. Melting temperature (Tm) of membranes increased with a decrease in endothermic peak with increasing beta chitosan content. The tensile strength of membranes in the dry state was found to be high (29-43 MPa) and it increased with increasing beta-chitosan content. The in vitro haemorheological studies indicated the biocompatible nature of membranes with no significant changes in whole blood and plasma viscosity and red blood cell rigidity. PMID- 15348381 TI - The sintering and mechanical behavior of hydroxyapatite with bioglass additions. AB - There is increasing interest in the potential of composites of hydroxyapatite with phosphate- or silicate-based bioactive glasses, and certain of these glass additions have been found, in previous work, to aid densification and form a mechanically-reinforced, bioactive material; in particular, large improvements in flexural strength and fracture toughness were obtained through the addition of small amounts of phosphate glass. Less is known about the mechanical behavior of HA/bioglass composites, although in vivo studies by other workers have shown encouraging biological results. In this investigation, the sintering behavior, mechanical properties, and microstructure of composites of HA with up to 50 wt % glass, were analyzed. X-ray diffraction showed the phase composition of sintered composites with up to 5 wt % added bioglass to be non-stoichiometric HA with alpha-TCP or beta-TCP. Phase analysis of composites containing higher glass additions was impracticable due to peak broadening and overlap, although reaction products, at the highest glass additions and sintering temperatures, may include wollastonite-2M and beta-Na2Ca4(PO4)2SiO4. Sintered density, and mechanical properties other than fracture toughness, showed no significant improvement over HA. PMID- 15348382 TI - Characterization of the transformation from calcium-deficient apatite to beta tricalcium phosphate. AB - The structural changes that occur during the transformation of a Ca-deficient apatite, prepared by a wet chemical method, to beta-TCP were investigated. X-ray diffraction (XRD) analysis of as-prepared samples and samples calcined at temperatures between 500 and 1100 degrees C showed that the transformation occurs over the temperature range 710-740 degrees C, under non-equilibrium conditions. The change in crystallite size with increasing calcination/sintering temperature was studied by XRD using the Scherrer formula. Fourier transform infra-red (FTIR) analysis indicated considerable structural change in samples above and below this temperature range. Changes were observed in the hydroxyl, carbonate and phosphate bands as the calcination temperature was increased from 500 to 1100 degrees C. Even once a single beta-TCP phase is obtained at 740 degrees C there remains a considerable amount of structural change at temperatures between 740 and 1100 degrees C. This effect was illustrated by an unusual change in the lattice parameters of the beta-TCP structure and significant changes in the phosphate bands of the FTIR spectra as the calcination temperature was increased. The results obtained in this study show that the combined experimental techniques of XRD and FTIR are excellent complimentary methods for characterizing structural changes that occur during phase transformations. PMID- 15348383 TI - PolyHEMA and polyHEMA-poly(MMA-co-AA) as substrates for culturing Vero cells. AB - Poly (2-hydroxyethyl methacrylate), polyHEMA, is known to prevent cellular attachment and spreading. This hydrogel is used to culture cells not dependent on anchorage. Blending polyHEMA with a copolymer of methyl methacrylate and acrylic acid introduces negative charges to the hydrogel and improves its mechanical characteristics. PolyHEMA and the blend were tested for attachment and proliferation of Vero cells. Dense and porous samples of the hydrogels were used. Attachment assays included cellular quantification with MTT photometry and cellular morphology with the scanning electron microscopy after 2 h culture. Proliferation assays were carried out with 5 and 10 days culture. Cellular morphology included cytochemistry of resin sections and scanning electron microscope observations. Hydrogels allowed a few cells to attach and proliferate. The cells growing on the surface of hydrogels were organized in various layers and showed a differential morphology. Cells located inside the pores remained rounded. The hydrogels showed the possibility of inducing differentiated phenotypic expression. PMID- 15348384 TI - Effect of casting method on castability of titanium and dental alloys. AB - Titanium, once considered to be difficult to cast because of its relatively high melting point (1670 +/- 50 degrees C) and strong chemical affinity, can now be acceptably cast using newly developed casting apparatus. The objectives of this study were to examine the castability of commercially pure (CP) titanium using an ultra high-speed centrifugal casting machine and a pressure difference-type casting unit and to compare the castability of titanium with that of conventional dental casting alloys. To determine castability, two types of patterns were used: a mesh pattern of 22 x 24 mm cut polyether thread sieve, and a saucer pattern (24 mm diameter) perforated to create four T-shaped ends. The casting equipment significantly affected the mold filling of both patterns (p < 0.001). The castability indices obtained from both patterns of CP titanium cast in the centrifugal casting machine were significantly (p < 0.05) better than the indices of the castings produced in the pressure-difference casting unit. The radiographs of the saucer pattern cast in the centrifugal casting machine showed some pores that were fewer and smaller in size than the pores found in castings made in the pressure-difference unit. When the ultra high-speed centrifugal casting machine was used with the manufacturer's recommended mold material, the castability of titanium was similar to that of gold alloy or Ni-Cr alloy cast by conventional means. PMID- 15348385 TI - Formation of bioactive functionally graded structure on Ti-6Al-4V alloy by chemical surface treatment. AB - An Al- and V-free sodium titanate hydrogel layer with a graded structure where the sodium titanate gradually decreases toward the interior, was formed on the surface of Ti-6Al-4V alloy, when the alloy was exposed to 5M NaOH solution at 60 degrees C for 24 h. This gel layer was transformed into an amorphous sodium titanate layer without giving considerable change in the graded structure, except a little increase in the depth of the oxygen distribution by a heat treatment at 600 degrees C for 1 h. The sodium titanate layer formed Ti-OH groups on its surface by exchanging its Na+ ion with H3O+ ion in simulated body fluid when soaked in the fluid, and thus formed Ti-OH groups induced the apatite nucleation. The apatite layer also formed a graded structure toward the substrate. The strong bond of the apatite layer to the substrate was attributed to this graded structure. PMID- 15348386 TI - Conservation of the elastic and flexural moduli of osteopenic femoral cortical bone in experimental inflammatory arthritis in the rabbit. AB - Experimental inflammatory arthritis (EIA) produced by carrageenan injection provokes a rapid bone remodeling state with cortical and cancellous bone loss. The objective of this study was to determine whether changes in cortical mechanical properties and/or geometry occur in long bones, either near or remote to the site of inflammation. EIA was induced in the right tibio-femoral joint of rabbits over 56 days. The right humerus and right femur from 15 normal and 25 arthritis group animals were excized. Semi-cylindrical specimens of the medial cortical shaft were subjected to non-destructive four-point bending tests. Transverse sections at the four contact sites of the loading jig were photographed and digitized to obtain average cross-sectional area (A) and moment of inertia (I). Moment of inertia and slope of the load/deflection curve permitted calculation of modulus of elasticity (E) for each specimen. Load/time curves were also used to calculate per cent stress remaining in relaxation experiments. Per cent stress remaining, E, A, I and square root of I/A (radius of gyration) were examined for differences by bone (humerus, femur) and by treatment (N,A) using two way ANOVA. The induction of inflammatory arthritis did not significantly alter the modulus of elasticity in either the femur or humerus; however, arthritis reduced the moment of inertia from 34.54 +/- 2.88 x 10(-12) m4 to 25.06+/-1.80 x 10(-12) m4 (mean+/-SEM, p < 0.05). This was observed in the femur (near the arthritic joint), but not in the humerus (remote from arthritic joint). Analysis of area and ratio I/A demonstrated that this geometric effect of treatment was due to reduced area without gross cross-sectional shape changes. Per cent stress remaining in the femur (but not in the humerus) was higher in the arthritis specimens than in the normal specimens (N: 80.86 < 0.97%; A: 83.25 +/- 0.71%, p < 0.05). Thus, in this arthritis model, the principal mechanical or geometric effect on cortical bone was reduction of the cross-sectional area and moment of inertia. The viscoelastic relaxation response of bone was also altered, perhaps due to loss of water or collagen degradation. PMID- 15348387 TI - Modeling of damage to articulating surfaces by third body particles in total joint replacements. AB - Numerous small scratches and some larger scratches have been observed on metallic femoral heads of explanted hip prostheses, with the larger scratches believed to be a major contributor to increased wear of the polyethylene acetabular cups. Previous work in our group has shown that smaller scratches, with a mean lip height up to 0.35 microm, can be caused by bone cement and bone particles up to 500 microm in size. However, the larger scratches were not readily replicated with these particles. Therefore in this study experimental and theoretical models have been developed to investigate the damage caused by harder metallic and ceramic particles. Small 10 microm diameter spherical metallic particles were also found to produce small fine scratches on the metallic counterface. However larger diameter spherical metal particles greater than 100 microm in diameter, which were embedded in polyethylene pins, caused severe sharp scratching of the metallic counterface with scratch lips greater than 0.5 microm. This level of damage, which was comparable to the severe damage found in vivo, was also simulated by a three body finite element model. Thus the larger metal particles led to the type of damage which was predicted to increase wear dramatically. This technique for simulating severe in vivo third body damage using spherical metal particles was found to be reproducible and reliable and will be used in the future in hip simulator testing to replicate third body damage and wear. PMID- 15348388 TI - The effect of osteopenia on the osteointegration of different biomaterials: histomorphometric study in rats. AB - The osteointegration of Hydroxyapatite (HA), Titanium (Ti-6Al-4V: Ti), Zirconia (ZrO2), Alumina (Al2O3) and 2 biological glasses (AP40 and RKKP) was comparatively investigated in normal and osteopenic rats by means of histomorphometry. Thirty-six Sprague Dawley female rats were left intact (Group C) while 36 were ovariectomized (Group OVX). Group C and OVX were further divided into 6 subgroups. After 16 weeks all animals were submitted to the femoral implant of nails made of the above-mentioned materials. Eight weeks after implantation the animals were euthanized, the femurs were harvested for histomorphometric analysis. The data showed that: (1) all the tested materials were biocompatible in vitro; (2) no significant differences existed in Affinity Index (AI) of Group C; and (3) results from paired comparison applied to the AI showed significant differences among the Groups C and OVX. The AI did not significantly change among intact groups, while it significantly decreased when some materials were implanted in OVX subgroups (AP40, ZrO2 and Ti-6Al-4V: p < 0.0005, p < 0.05 and p < 0.01). It is confirmed that bone mineral density is a strong predictor of the osteointegration of an orthopedic implant and that the use of pathological animal models is necessary to completely characterize biomaterials. PMID- 15348389 TI - Biocompatibility of microplates for culturing epithelial renal cells evaluated by a microcalorimetric technique. AB - In the present study we have developed a microcalorimetric procedure which allows convenient investigation of biocompatibility in a microsystem. We examined the biocompatibility of a porcine renal epithelial tubule cell line LLC-PK1 and a human primary renal epithelial tubule cell (RPTEC) with microplates composed of three different materials, i.e. Thermanox, transparent film and titanium. All three materials showed equal biocompatibility with LLC-PK1 cells, judging from the attainment of steady-state power curves and the same rate of heat production per cell (2.5 microW / microg DNA). The human renal cells were poorly biocompatible with the Thermanox and transparent film. However, on titanium the RPTEC cell did adhere, as demonstrated by a steady-state power curve. The human cells also showed a higher metabolic activity (3.0 microW / microg DNA), than did LLC-PK1 cells cultured on the same type of microplates. In research on biocompatibility there is a need for alternatives to experimental animal investigations. The present technique allows studies of cellular interactions with different biomaterials in a rapid and standardized manner and may therefore prove to be a useful screening procedure. PMID- 15348390 TI - Radiation-induced grafting of methylmethacrylate onto ultrahigh molecular weight polyethylene and its adhesive characteristics. AB - The surface of ultrahigh molecular weight polyethylene (UHMWPE) was modified by radiation-grafting methylmethacrylate (MMA) in the presence of sulfuric acid and metallic salt to increase bonding strength with polymethylmethacrylate. The effect of the addition of metallic salts and sulfuric acid on the radiation grafting reaction was investigated when MMA was grafted to the irradiated UHMWPE. The adhesive characteristics with the grafting yield were investigated using conventional acrylic bone cement based on poly(methyl methacrylate) [PMMA]. The results showed that the inclusion of an FeSO4 . 7H2O and sulfuric acid in MMA grafting solutions was extremely beneficial and led to a most unusual synergistic effect, while CuSO4 . 5H2O led to a detrimental effect. The tensile bonding strength between UHMWPE and PMMA sheet increased remarkably with an increased grafting yield on UHMWPE surfaces. PMID- 15348391 TI - The effect of particle size and electrical charge on macrophage-osteoclast differentiation and bone resorption. AB - In aseptic loosening, there is commonly periprosthetic bone loss and a heavy macrophage infiltrate in response to biomaterial wear particles generated from the implant materials. Macrophages which have phagocytosed wear particles are known to be capable of differentiation into bone resorbing osteoclasts. In this investigation we determine the role of particle size and particle charge on this process. Mouse monocytes and macrophages were co-cultured with osteoblast-like UMR106 cells and 1,25 dihydroxyvitamin D3 in the presence or the absence of (i) various sizes of latex beads (0.1, 1, 10 and 100 microm) and (ii) uncharged, positively- or negatively-charged sephadex beads of uniform shape and composition. The extent of osteoclast differentiation by monocytes or foreign body macrophages was determined by the expression of the osteoclast-associated enzyme tartrate-resistant acid phosphatase and lacunar bone resorption. No significant difference in the extent of osteoclast formation and bone resorption was noted in response to particle size. Osteoclast formation was also not significantly different in the presence of positively/negatively charged and uncharged particles. These findings indicate that osteoclast formation is not significantly influenced by particle characteristics, such as particle size. They also add support to the hypothesis that macrophage involvement in periprosthetic osteolysis is not dependent on particle phagocytosis and that it may be induced by particle contact. PMID- 15348392 TI - Osseous regeneration in the presence of oxidized cellulose and collagen. AB - Oxidized cellulose and collagen are two absorbable hemostatic scaffolding materials that are used widely in surgery. A histomorphological study was undertaken to determine the tissue response and extent of healing brought about by intraosseously implanting these two materials in the femur and tibia of sheep. There was no major difference in the rate of repair of the bone defects brought about by these two materials, with the bone defects being completely repaired by lamellar bone at 6-8 weeks. Therefore, our results suggest that, in most instances where collagen is presently used in surgical applications, it could be substituted by oxidized cellulose. PMID- 15348393 TI - Effect of molecular weight and concentration of poly(acrylic acid) on the formation of a polymeric calcium phosphate cement. AB - Previous investigations have noted that the tetracalcium phosphate (TTCP)/dicalcium phosphate anhydrous (DCPA) apatite forming calcium phosphate cement (CPC) possesses many favorable properties from a biomaterials standpoint. Despite these positive properties various shortcomings have limited clinical usage of these materials and fostered investigations into the effect of numerous additives. The present study concerns the effect of poly(acrylic acid) (PAA) addition and the influence of factors such as molecular weight and concentration of the additive on the properties of the set cement. One-way ANOVA was conducted using all results obtained, to firstly derive the influence of concentration within each molecular weight group, and secondly to derive the influence of molecular weight within each concentration group. All investigated mechanical properties were influenced by both molecular weight and concentration of the additive. Higher molecular weights tended to result in cements with shorter setting times and higher compressive, diametral and biaxial flexural strengths than their lower molecular weight counterparts. The effect of concentration on the properties of the set cement however was somewhat more complex, a negative correlation was observed between the initial setting time and PAA concentration. In regards to the final setting time, any correlation with concentration was difficult to derive as a consequence of the highly brittle nature of cements made with low concentrations. In regard to mechanical properties, intermediate concentrations tended to give higher strengths than both their higher and lower counterparts, however the exact pattern was largely specific to the mechanical strength test employed. We conclude that molecular weight and concentration of PAA influence the setting behavior and final mechanical properties of the TTCP/DCPA cement, and that selection of an appropriate PAA solution can lead to the production of cements with properties superior to those formed in the absence of the polymer. PMID- 15348394 TI - Water sorption and water-induced molecular mobility in dental composite resins. AB - Water sorption in two resin composites, Kulzer's Solitaire (S) and SDI's Wave (W), and in a polyacid-modified composite resin, 3M's F2000 (compomer F), was investigated by means of equilibrium sorption isotherms (ESI) and of dynamic sorption (DS) measurements. Molecular mobility in these materials was studied by means of dielectric relaxation spectroscopy (DRS) and of thermally stimulated depolarization currents (TSDC) measurements. The results of ESI measurements show that at equilibrium, water is molecularly distributed in the materials and the effects of hydrophilic sites and clustering are negligible. Hysteresis effects in sorption-desorption cycles are larger in the resin composites than in the compomer. Equilibrium water uptakes in both ESI and DS conditions are rather low, in the range 1-2%. Diffusion coefficients of water are about 1x10(-8) cm(2)/s in the resin composites and by a factor of about 2 smaller in the compomer. Molecular mobility increases with hydration, as suggested by preliminary DRS and TSDC measurements. Detailed dielectric measurements may give important information for understanding, at the molecular level, water-induced degradation in dental materials. PMID- 15348395 TI - Calcium phosphate formation on nanocrystalline ZrO2 thin film prepared by using a zirconium naphthenate. AB - To investigate the calcium phosphate forming ability of ZrO(2) thin film, we prepared ZrO(2)/Si structure by a chemical solution deposition with a zirconium naphthenate as a starting material. Precursor sol was spin-coated onto the cleaned Si substrate and prefired at 500 degrees C for 10 min in air, followed by final annealing at 800 degrees C for 30 min in air. Surface morphology and surface roughness of the annealed layer were characterized by field emission scanning electron microscope and atomic force microscope. After soaking for 5 days in a simulated body fluid, formation of the calcium phosphate on nanocrystalline ZrO(2) layer annealed at 800 degrees C was observed by energy dispersive X-ray spectrometer. Fourier transform infrared spectroscopy revealed that carbonate was substituted into the calcium phosphate. PMID- 15348396 TI - Physical properties of high molecular weight 1,3-trimethylene carbonate and D,L lactide copolymers. AB - High molecular weight statistical copolymers of 1,3-trimethylene carbonate (TMC) and D,L-lactide (DLLA) were synthesized and characterized with the aim of assessing their potential in the development of degradable and flexible materials for application in the biomedical field. Under the applied polymerization conditions (130 degrees C, 3 days using stannous octoate as a catalyst) monomer conversion was high or almost complete, and high molecular weight polymers (M(n) above 170 000) were obtained. Significant improvement of the mechanical performance of these materials was observed in comparison to results previously reported for TMC and DLLA based copolymers of lower molecular weight. For the entire range of compositions the polymers are amorphous with a glass transition temperature ranging between -17 degrees C for poly(TMC) and 53 degrees C for poly(DLLA). The polymers vary from rubbers to stiff materials as the content of TMC decreases. All polymers are hydrophobic with very low equilibrium water absorption (<1.5 wt %). Thermal analyses and tensile tests were performed on polymer samples after water uptake. Due to a plasticizing effect of the water, the thermal properties, and consequently the mechanical performance, of the copolymers with higher content of DLLA were the most affected. After water absorption, the polymer mechanical behavior can change from glassy to rubbery, as observed for the copolymer with 80 mol % of DLLA. The obtained results suggest that these copolymers are promising candidates as biomaterials in the preparation of degradable medical devices and systems. PMID- 15348397 TI - Surface properties and in vitro analyses of immobilized chitosan onto polypropylene non-woven fabric surface using antenna-coupling microwave plasma. AB - Antenna coupling microwave plasma enables a highly efficient and oxidative treatment of the outermost surface of polypropylene (PP) non-woven fabric within a short time period. Subsequently, grafting copolymerization with acrylic acid (AAc) makes the plasma-treated fabric durably hydrophilic and excellent in water absorbency. With high grafting density and strong water affinity, the pAAc grafted fabric greatly becomes feasible as an intensive absorbent and as a support to promote chitosan-immobilization through amide bonds. Experimental result demonstrated that surface analyses by FTIR-ATR have shown that R-CONH-R', amide binding were emerged between pAAc and chitosan. The XPS measurements on C(1s) 286.0 eV (C-OH), 286.5 eV (C-N) and 288.1 eV (O=C-NH) also could be found. Bioactivity assessments on the chitosan-immobilized surfaces were anticipated by activated partial thromboplastin time (aPTT), thrombin time (TT), and fibrinogen concentration. By means of cell counter we counted the ratio of blood cell adhesion on the modified fabric matrix. After human plasma incubated with the chitosan-immobilized PP fabrics, the required time for aPTT and blood cell adhesion increased significantly, while fibrinogen concentration and TT did not change. Due to the capability of anticoagulation and cell adhesion, the chitosan immobilized PP fabric can be used as the substrate for cell culturing and then developed the wound-dressing substitute for second-degree burn. PMID- 15348398 TI - Rheological properties of poly(2-hydroxyethyl methacrylate) (pHEMA) as a function of water content and deformation frequency. AB - Poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogels have been used, or suggested for use, in a wide range of biomedical applications. In many of these applications, the mechanical properties of the gel are important for its proper functioning. These properties are influenced by a number of factors, including water content. In this study the storage and loss shear moduli were measured as a function of frequency for gels with water contents ranging from 22% to 48% at a temperature of 37 degrees C. At low frequencies and high water contents, deformation frequency had little effect. However, at higher frequencies and lower water contents, both moduli increased markedly with increasing frequency. This can be explained by the gels approaching a glass transition. The curves describing the behavior of each gel were combined to form a master curve, using a method analogous to the time-temperature superposition principle. This master curve can be used to predict the shear moduli for gels with a wide range of water contents and loading frequencies. For example, for a gel with a water content of 47.8% (as a percentage of the mass of gel), the curve provides shear moduli values over a frequency range of 10(-2)-10(4) Hz. PMID- 15348399 TI - Effect of crosslinking, thermal treatment and UV irradiation on the mechanical properties and in vitro degradation behavior of several natural proteins aimed to be used in the biomedical field. AB - Gelatine (GEL), soy (SI), casein (CAS) and sodium-caseinate (NaCAS) solutions were cast to produce protein films. All the proteins were chemically modified by adding glyoxal to the film-forming solutions in amounts varying from 0 to 0.9% (w/w based on the protein content). After casting, the same films were also submitted to a heat treatment performed at 80 degrees C or UV irradiation. The effect of those chemical/physical modifications on the mechanical properties and on the hydrolytic stability of the protein films was evaluated. As a result, a large variety of protein films with different mechanical properties and degradation profiles were developed. CAS and NaCAS even when chemically/physically modified do not resist to hydrolysis longer than 2 weeks. GEL, only when chemically modified with glyoxal, become water resistant. Due to its hydrolytic stability, SI become a very attractive material for biomedical applications where long term treatments are a requisite. PMID- 15348400 TI - Colloidal characterization and electrophoretic deposition of hydroxyapatite on titanium substrate. AB - Hydroxyapatite (HA) powders were prepared by a modified chemical co-precipitation method and electrophoretically deposited onto a titanium tubular substrate. The zeta potential, electromobility and the particle size of the HA suspension was characterized at various pH values and the most stable and dispersed suspension condition was identified. Electrophoretic deposition of the HA particles on the titanium substrate was then carried out at this optimum suspension condition. Studies on deposition rate and examination on the microstructure of the sintered deposit were performed. The stoichiometry of the HA before and after sintering were also confirmed. The deposition experimental data obtained in the present work was also compared with theoretical model proposed in the literature. Lastly, the adhesion strength of the coating was also quantified using shear strength tests. PMID- 15348401 TI - The semi-interpenetrating polymer network matrix of fiber-reinforced composite and its effect on the surface adhesive properties. AB - This aim of this study was to examine the effect of further-impregnation time of polymer pre-impregnated fiber-reinforcement on polymer matrix structure of the fiber-reinforced composite (FRC) used in dental applications. In addition, shear bond strength between the FRC and veneering composite after various length of further-impregnation was studied. Polymethyl methacrylate (PMMA) pre-impregnated glass fiber-reinforcement was further-impregnated with a diacrylate monomer resin by using five lengths of further-impregnation from 10 min to 24 h. The test specimens (n=5) from each five groups were treated with the solvent tetrahydrofuran and examined with a scanning electron microscope (SEM) to determinate the existence of linear PMMA in the polymer matrix of the FRC. The same lengths of further-impregnation were used to form an adhesive substrate for veneering composite and to measure the shear bond strength (n=8). The SEM examination showed that linear PMMA-polymer and cross-linked diacrylate polymer formed two independent networks for the polymer matrix of FRC. The highest mean shear bond strength value (18.7+/-2.9 MPa) was achieved when the fiber reinforcement was further-impregnated for 24 h. The shortest further impregnation, 10 min, resulted in the lowest mean shear bond strength (12.7+/-2.9 MPa). A correlation between increased shear bond strength and longer further impregnation was found (0.689, p<0.001). The results revealed that linear PMMA network of the polymer matrix of the FRC remained in the structure regardless of the various lengths of the further-impregnation with diacrylate resin. PMID- 15348402 TI - Stress relaxation tests in polypropylene monofilament meshes used in the repair of abdominal walls. AB - The objective of this work has been to characterize stress relaxation in the polymer material on applying different levels of constant strain. The meshes were strained at values of 5.2%, 5.4% and 5.6% which are the values at which the mesh is strained in clinical use for the repair of abdominal walls. Laws have been obtained to model the viscoelastic behavior at different strains for this material. Finally, fracture studies were carried out by environmental scanning electron microscopy to determine the fracture mechanisms of these meshes. Besides, the implantation of the meshes was practised in two different layers of abdominal wall: the superficial or preaponeurotic layer and deep or preperitoneal layer, showing the neoformation of connective tissue on the mesh, which tended to be organized differently in each layer studied; more roughly and densely in the superficial layer than in the deep one. PMID- 15348403 TI - Effects of sintering temperature on structure of hydroxyapatite studied with Rietveld method. AB - It is obvious that the sintering temperature of hydroxyapatite (HA) ceramics significantly affects their biological responses in vitro or in vivo. The HA ceramics sintered at different temperatures exhibit a wide variation of biological response, but the correlation of this variation with the parameters of HA crystalline structure is not fully investigated. In present study, the crystalline structure of HA powders sintered at different temperatures at 600, 800, 1000, 1200 degrees C, was characterized using X-ray diffraction (XRD), and then refined with Rietveld method. A series of structure parameters, such as, cell lattice parameters (a and c) and bond length, distances of special atoms on surface to nearest atom, a numerical index of distortion for PO(4) tetrahedron, as well as internal energy and density of HA cell, were calculated to characterize the crystalline structure of HA at atom level. The broadening effect of XRD reflections was also separated to calculate the micro-strain/crystalline size, respectively. These parameters of HA crystalline structure and cell demonstrated that with rising of the sintered temperature, the internal energy and micro-strain of HA cell decreased, but the crystalline size increased. The regularity of PO(4) tetrahedron and distance between Ca(1) and O(3) in HA cell showed the same tendency as the internal energy and micro-strain with sintering temperature. All of these parameters indicated that HA became more stable with the rising of sintering temperature. PMID- 15348404 TI - Journal of materials science: materials in medicine. PMID- 15348405 TI - Protein adsorption and fibroblast adhesion on irradiated polysiloxane surfaces. AB - A very peculiar case of differential cell response towards polysiloxane surfaces of very similar composition is investigated. Poly(hydroxymethylsiloxane) (PHMS) surfaces treated either by O(2)-plasma or 6 keV Ar(+)-beams have been used to test the adhesion, proliferation and spreading of human fibroblasts. The surface chemical structure and nanomorphology were investigated by means of X-ray photoelectron spectroscopy (XPS), surface free energy measurements and atomic force microscopy (AFM). In spite of the close compositional and morphological similarity of the modified surfaces, the viability of the adhered cells, evaluated by means of optical microscopy and epifluorescence microscopy, was found to be very different in the two cases. The study of the features of the adsorbed protein adlayer on the two types of surfaces was performed by XPS and AFM and indicated that the overall cell behavior is connected to a quite different protein aggregation process, occurring respectively on the plasma- and Ar(+)-modified polysiloxane surfaces. It is suggested that the specific biological response of the modified surfaces is determined by the chemical structure at the nanometric level. PMID- 15348406 TI - Effects of adsorbed heat labile serum proteins and fibrinogen on adhesion and apoptosis of monocytes/macrophages on biomaterials. AB - A previously established human monocyte culture protocol was used to determine the effects of varying adsorbed proteins on monocyte/macrophage adhesion and survival on dimethyl-silane (DM) or RGD modified glass coverslips. Cells were allowed to adhere for 2 h in the absence of protein or in the presence of serum, fibrinogen (Fg), heat inactivated serum (HIS), serum supplemented with Fg or HIS with Fg. Cell adhesion and apoptosis rates were determined on days 0 (2 h), 3, 7 and 10 of culture. The presence of serum alone in the initial culture was sufficient to optimize monocyte/macrophage adhesion and survival rates. Adding Fg to serum did not increase adhesion nor decrease apoptotic rates. No protein or the addition of HIS during the initial incubation period significantly decreased monocyte/macrophage adhesion and survival on both surfaces, however, the addition of Fg to HIS restored adhesion and survival rates to those seen with in the presence of serum alone on RGD surfaces. These studies demonstrate that monocyte/macrophage adhesion and survival on biomaterial surfaces are optimized by adsorbed heat labile serum proteins while adsorbed Fg plays a surface property dependent role. PMID- 15348407 TI - Experimental approaches to study vascularization in tissue engineering and biomaterial applications. AB - The success of tissue engineering and biomaterial applications is not only dependent on the growth and functioning of the organ- or tissue-specific cells on the biomaterial but is entirely dependent in most cases on a successful vascularization after implantation. The process of vascularization involves angiogenesis; the formation of new blood vessels which spread into the implant material and supply the existing cells with the nutrients to survive. We have established in vitro methods using human microvascular endothelial cells to evaluate novel biomaterials for endothelial cell attachment, cytotoxicity, growth, angiogenesis and the effects on gene regulation. These in vitro studies can be used to rapidly evaluate the potential success of a new biomaterial and for the development of matrix scaffolds which will promote a physiological vascularization response. PMID- 15348408 TI - Design, synthesis and properties of polyurethane hydrogels for tissue engineering. AB - Due to their similarity to natural soft tissues, water-swellable polymeric materials (hydrogels) are, in principle, ideal candidates for scaffolds/matrices in tissue engineering. Polyurethanes (PU), hydrophilic but water-insoluble, can be obtained by the incorporation of hydrophilic soft segments, e.g. poly(ethylene oxide) (PEO). These materials possess the favorable characteristics of the family of PUs as well as the ability to mimic soft tissues. In this work, new crosslinked PU-hydrogels were prepared in a one-step bulk polymerization process using an aliphatic diisocyanate, PEO, a low molecular weight diol, and a tri functional crosslinking agent. A porous structure was also obtained by air incorporation under mechanical stirring at a controlled high speed during the polymerization. Structural characteristics of the compact (PU-HyC) and the porous (PU-HyP) material were investigated. Molecular weight between cross-links, M(c), and crosslinking density, rho(x), were typical of a low crosslinking degree. A homogeneous distribution of non-interconnecting pores (phi100 microm) was observed in PU-HyP. Both materials showed a high water adsorption. The swelling behavior and weight loss in water was affected by porosity. For their mechanical behavior in the swollen state, the novel PU hydrogels can be considered for biomedical applications where good mechanical properties are required (i.e. 3D scaffold for tissue engineering). PMID- 15348409 TI - Poly(vinyl alcohol) as versatile biomaterial for potential biomedical applications. AB - In this paper, we present some new case examples where the chemical versatility of poly (vinyl alcohol) (PVA) can be used for potential biomedical applications. PVA, the polymeric material used for designing new nanostructured devices, is water soluble, biocompatible and has excellent physical properties. We point out the possibility of obtaining wall-to-wall chemical hydrogels as well as microgels without diminishing the biocompatibility available in the starting PVA material. Injectability is another important factor to take into account in controlled drug delivery for gene therapy. In this respect, in this paper, established and more innovative methods are prospected in order to obtain particles with dimensions suitable for these applications. PMID- 15348410 TI - Enhanced HAPEX topography: comparison of osteoblast response to established cement. AB - The use of poly(methylmethacrylate) PMMA cement by Charnley in the 1960s revolutionized orthopaedic medicine. Since this time, however, little has changed. The development of bioactive composites, such as HAPEX (a composite of 40% vol hydroxyapatite (HA) in a polyethylene matrix) have potential in orthopaedic applications. The composite has been shown to allow direct bone bonding in vivo, and in vitro studies have shown preferential attachment to HA exposed on the composite surface. In vitro study has also shown that altering the topography HAPEX can enhance osteoblast response. This study uses microscopical investigation of osteoblast cytoskeleton, and biochemical measurement of proliferation (by thymidine incorporation) and phenotype (by alkaline phosphatase activity) to compare primary human osteoblast (HOB) activity on HAPEX and PMMA cement. The study shows large increases in HOB response to the new generation material compared to PMMA, the current implant standard. PMID- 15348411 TI - "Smart" polymeric nanospheres as new materials for possible biomedical applications. AB - Novel random terpolymers of N-isopropylacrylamide (NIPAM), sodium 2-acrylamido-2 methyl-1-propanesulfonate (AMPS), and cinnamoyloxyethylmethacrylate (CEMA) were synthesized by free radical copolymerization using AIBN as an initiator. Five terpolymers were obtained by copolymerization of the monomer mixtures containing a fixed amount of 10 mol % of AMPS while the content of CEMA ranged from 5 to 25 mol % and was changed in 5 mol % increments. The terpolymers obtained are water soluble. Because of their amphiphilic nature they undergo self-organization in the aqueous solution with the formation of micelles capable of solubilizing sparingly water soluble organic compounds, such as drugs. The terpolymers are susceptible to three external stimuli, i.e. temperature, ionic strength and UV light. Due to the presence of NIPAM in the terpolymers they display the lower critical solution temperature (LCST), the presence of AMPS makes them sensitive to the ionic strength of the solution, while the light-responsiveness of the terpolymers is due to the presence of cinnamoyl chromophores, which undergo photodimerization when irradiated with UV light at about 280 nm. Application of any of these stimuli alone or in combination with other stimuli allows changing the copolymer properties in a controlled way. PMID- 15348412 TI - Nanoparticle systems for the targeted release of active principles of proteic nature. AB - The preparation and characterization of nanoparticles based on biodegradable/bioerodible polymers is reported. They have been designed for the controlled-targeted release of proteic drugs such as alpha-interferon and for the release of active principles in tissue engineering. The amenability of some of the prepared polymeric matrices to be used in the fabrication of micro and nano patterned scaffolds is also described. PMID- 15348413 TI - Engineered articular cartilage: influence of the scaffold on cell phenotype and proliferation. AB - Articular cartilage defects do not heal. Biodegradable scaffolds have been studied for cartilage engineering in order to implant autologous chondrocytes and help cartilage repair. We tested some new collagen matrices differing in collagen type, origin, structure and methods of extraction and purification, and compared the behavior of human chondrocytes cultured on them. Human chondrocytes were grown for three weeks on four different equine type I collagen matrices, one type I, III porcine collagen matrix and one porcine type II collagen matrix. After 21 days, samples were subjected to histochemical, immunohistochemical and histomorphometric analysis to study phenotype expression and cell adhesion. At 7, 14 and 21 days cell proliferation was studied by incorporation of [3H]-thymidine. Our data evidence that the collagen type influences cell morphology, adhesion and growth; indeed, cellularity and rate of proliferation were significantly higher and cells were rounder on the collagen II matrix than on either of the collagen I matrices. Among the collagen I matrices, we observed a great variability in terms of cell adhesion and proliferation. The present study allowed us to identify one type I collagen matrix and one type II collagen matrix that could be usefully employed as a scaffold for chondrocyte transplantation. PMID- 15348414 TI - Effects of elastin-derived peptide on Achilles' tendon healing: an experimental study. AB - Different matrix macromolecules modulate the tendon healing process. Elastin contains sequences which exhibit chemotactic activity both in vitro and in vivo. We analyzed the effects of synthetic elastin-derived peptide Val-Gly-Val-Ala-Pro Gly suspended in a gel solution on the healing process of Achilles' tendon in a rat model. A total tenotomy at the middle 3rd was performed in 32 rats. During the suture repair the gel with (Group A) or without (Group B) the elastin-derived peptide was applied to the tendon stumps. Four animals for each period and group were killed at 10, 30, 60 and 90 days after surgery. The scar tissue was processed for histochemical, immuno-histochemical and morphometric analysis. An improved healing process with increase in cellularity and vascularity, especially at the early stage of the Achilles' tendon healing process was observed in Group A compared to Group B. The fiber alignment was also positively influenced by the factor. Immunolabeling with HAM 56 and lisozyme revealed a stronger reaction for the presence of monocyte/macrophage in Group A vs Group B especially in early stages. Chondral metaplasia and endochondral ossification occurred in the healed tissue of both group at 60 and 90 days. PMID- 15348415 TI - Micro and nano-structured surfaces. AB - The study of cell reaction to micro and nanotopography is dependent on the method of manufacture available. Several methods of manufacture have been developed: polymer demixing, embossing and photolithography. Surfaces obtained with these different techniques, having micro and/or nanodomains, have been studied toward the same type of cells, i.e. human endothelial cells (HGTFN) and mouse fibroblasts (3T3). Polymer demixing of polystyrene (PS) and poly(4-bromostyrene) (PBrS) producing nanometrically islands of 18, 45 and 100 nm height, polycarbonate (PC) and polycaprolactone (PCL) grooved with grooves 450 nm wide and 190 high, the natural polysaccharide hyaluronic acid (Hyal) and its sulfated derivative (HyalS) photoimmobilized on silanized glass as grooves 250 nm high and 100, 50, 25 or 10 microm wide have been obtained. The morphology and polarization of the cells has been studied by optical microscopy and scanning electron microscopy. Cells respond in different way to the topography of the materials, but the surface chemistry is dominant in inducing different cell behavior. PMID- 15348416 TI - Gland cell cultures into 3D hyaluronan-based scaffolds. AB - In this study we report a preliminary investigation of the feasibility of non woven/sponge fabrics of a hyaluronan derived biomaterials (benzyl ester of HA (HYAFF-11 FAB, Abano Terme, Italy) for the in vitro culture of rat hepatocytes and rat beta cells. Cell growth on hyaluronan derived biomaterials were tested in the presence of complete medium and in the presence of ECM (extracellular matrix) secreted by fibroblasts previously cultured into the scaffold. Hepatocytes and beta cells were extracted from rat liver/pancreas and seeded either on the HYAFF 11 scaffold alone, or on HYAFF-11 scaffold containing ECM. Direct assay of cell proliferation was performed with MTT test. For morphological observations samples were stained with hematoxylin and eosin. The results obtained by MTT test showed that hepatocytes cultivated in both the above described conditions were able to proliferate up to 14 days and Langerhans islet up to 21 days. After this time, cells started to undergo apoptosis. The morphological analyses showed cell aggregation in three-dimensional structures promoted by the fibers of the biomaterial. Our results confirmed that HYAFF-11 meshes represent a suitable scaffold for hepatocyte adhesion/Langerhans islet organization and proliferation. In particular, the presence of a fibroblast secreted extracellular matrix improves the biological property of the scaffold. PMID- 15348417 TI - Apatite deposition on polyamide films containing carboxyl group in a biomimetic solution. AB - The development of organic-inorganic hybrids composed of hydroxyapatite and organic polymers is attractive because of their novelty in being materials that show a bone-bonding ability, i.e. bioactivity, and because they have mechanical properties similar to those of natural bone. The biomimetic process has received much attention for fabricating such a hybrid, where bone-like apatite is deposited under ambient conditions on polymer substrates in a simulated body fluid (SBF) having ion concentrations nearly equal to those of human extracellular fluid or related solutions. It has been shown that the carboxyl group is effective for inducing heterogeneous nucleation of apatite in the body. In the present study, apatite deposition on polyamide films containing various numbers of carboxyl groups was investigated in 1.5 SBF, which had ion concentrations 1.5 times those of a normal SBF. The effect of incorporation of calcium chloride on the formation of apatite was examined. Polyamide films containing or=40 mass % CaCl(2) formed apatite on their surfaces in 1.5 SBF. The ability of the modified film to form an apatite layer increased, and the adhesion of the apatite layer bonded to the film improved, with increasing carboxyl group content. It is concluded that novel apatite-polyamide hybrids can be prepared by a biomimetic process. PMID- 15348418 TI - A novel acrylic copolymer for a poly(alkenoate) glass-ionomer cement. AB - The interest in the clinical use of polyalkenoate cements stems mainly from their behavior as bioactive adhesive materials with therapeutic action. Glass-ionomer cements set by an acid-base reaction between a degradable glass and a poly(alkenoic acid) and the therapeutic action is related to the release of fluoride ions which are present in the hardened cement that show a sustained release over years, responsible for caries inhibition in teeth. Conventional glass-ionomers, however, suffer from some disadvantages such as short working time, initial moisture sensitivity and prone to desiccation after setting and are generally brittle. In the present study, a poly(alkenoic acid) copolymer was synthesized based on acrylic acid and 2-hydroxyethylmethacrylate (HEMA) using azobisisobutyronitrile as the initiator and characterized. The acid-base reaction was carried out by reacting aqueous solutions of the new copolymer (40 and 60%) with a commercial aluminofluorosilicate glasses as used in conventional glass ionomer cements. The results showed that the copolymer of HEMA and acrylic acid was a viable poly(alkenoic) acid for formation of glass-ionomer cements. PMID- 15348419 TI - Dynamic mechanical behavior of PMMA based bone cements in wet environment. AB - The mechanical properties of three wet commercial bone cements, namely Braxel (from Bioland), Simplex-P (from Howmedica) and CMW1-G (from DePuy) are investigated by means of stress relaxation and dynamic mechanical analysis (DMA). The geometry of loading that was used is the three point bending method (ASTM D790); all the tests were performed in a water chamber by means of temperature sweeps between 17 and 57 degrees C and spanning four frequency decades. The results show that viscoelastic properties are strongly dependent on specimen conditioning (i.e. water uptake and heat treatment). The results also show that all the cements that were analyzed show mechanical properties which are intermediate between the ones of the cancellous bone and of the metals of which prostheses are normally made. As a consequence, the cement is able to reduce the stress concentrations due to the interfacing of materials which have very different stiffnesses. Moreover, the results of the DMA, particularly the ones concerning the damping factor (tan delta), indicate that at body temperature the bone cements tested show an increased capacity of dissipation, the higher is the loading frequency, thus displaying shock absorbing properties. PMID- 15348420 TI - Cytotoxicity study of homopolymers and copolymers of 2-hydroxyethyl methacrylate and some alkyl acrylates for potential use as temporary skin substitutes. AB - The cytotoxicities of both homopolymers and copolymers of 2-hydroxyethyl methacrylate (HEMA) and various alkyl acrylates (AA) were investigated using cell culture techniques. The particular alkyl acrylates used were methyl acrylate (MA), ethyl acrylate (EA) and butyl acrylate (BA). The AA contents of the polymers studied were in the range of 0-25% by weight. They were synthesized via bulk polymerization in the form of thin sheets of 0.5+/-0.1 mm thickness. After sterilization using ethylene oxide gas, cytotoxicity tests, including both direct and indirect contact tests, were performed using cell lines of L929 and normal human dermal fibroblasts. The results showed that P(HEMA-co-EA) with EA contents of 15-25% and P(HEMA-co-BA) with BA contents of 10-25% appeared to be non cytotoxic and attractive for more specific cytocompatibility tests and in vivo studies with a view to their use as temporary skin substitutes. PMID- 15348421 TI - Ion-release, dissolution and buffering by zinc phosphate dental cements. AB - The interaction of zinc phosphate dental cement with aqueous solutions has been studied in order to elucidate the relationship between pH change and ion release (dissolution). For each storage medium (deionized water, lactic acid at pH 2.7 and lactate buffer at pH 2.2) five cylindrical specimens of zinc phosphate cement (6 mm diameter x 12 mm height) were prepared and weighed. They were stored individually in 8 cm(3) of solution for a week, then the pH was determined and the specimens reweighed. The solutions were replaced and the specimens stored for a further week, then the pH and the weight were again measured. This was repeated for four weeks. For each storage solution at each time interval, the concentration of ions leached (Na, Mg, Al, Zn and P) were determined using ICP OES. The lactate buffer was particularly erosive and reduced specimens to 4.1% (+/-0.9%) of their original mass after 4 weeks. The lactic acid was also erosive, but in water, specimens showed no significant mass change after 4 weeks. In all media, Na, Al, Mg, Zn and P ions were released, with mole ratios varying at each time interval. In all cases, the pH shifted towards neutral, but the relationship between ion release and solution pH was not straightforward. From the mole ratios of ions, estimates could be made of the relative proportions of attack at matrix to attack at filler, and this showed attack at filler predominated in most solutions at most time intervals. PMID- 15348422 TI - Amorphous alloys resistant to corrosion in artificial saliva solution. AB - The tailoring of new corrosion-resistant alloys with specific properties has recently been performed mostly by the sputter deposition technique. The aim of this work was to investigate corrosion resistance of aluminum-tungsten (Al-W) amorphous alloys in artificial saliva solution, pH=5.5, based on the electrochemical methods of cyclic voltammetry and linear polarization. Thin alloy films were prepared on a sapphire substrate by magnetron codeposition. Completely amorphous films were obtained in the Al(80)W(20)-Al(67)W(33) composition range. Amorphous Al-W alloys exhibit very high corrosion resistance due to their homogeneous single-phase nature. The passive films spontaneously formed at their surface are uniform with characteristics of an insulator film and prevent corrosion progression in the bulk in a very demanding oral environment. The mechanism of increasing resistivity of Al-W alloys to pitting corrosion and generalized corrosion has been discussed in the view of increasing tungsten content in the alloy. Considering these exceptional corrosion properties and microhardness which falls in the range 7.5+/-1.6 Pa, Al-W alloys represent promising materials for dental applications. PMID- 15348423 TI - Grafting of lactose-carrying styrene onto polystrene dishes using plasma glow discharge and their interaction with hepatocytes. AB - Lactose-carrying styrene (VLA)-grafted polystyrene (PS) dish (PS-VLA) was prepared by treatment of PS dish with oxygen plasma glow discharge followed by the graft polymerization of VLA. The surface topology and hepatocytes behavior on PS-VLA were examined by comparison with those on a PVLA-coated PS dish (PS-PVLA). According to the results of surface topologies obtained by a phase mode of atomic force microscope (AFM), it was found that PS-VLA exhibits a pointed texture image similar to forest while PS-PVLA exhibits a phase-separated, cloud-like image. In an experiment involving hepatocytes adhesion, the cells more slowly adhered to PS VLA than to PS-PVLA during the first 2 h incubation. According to topological data, it may be suggested that lactose density on the air side surface of PS-VLA is lower than that of PS-PVLA, thus leading to the slow adhesion of hepatocytes to PS-VLA. PMID- 15348424 TI - A metastable phase in thermal decomposition of Ca-deficient hydroxyapatite. AB - We investigated the microstructural changes on an atomic length scale during thermal decomposition process of Ca-deficient hydroxyapatite (Ca-def HAp) by high resolution transmission electron microscopy (HRTEM). Ca-def HAp was prepared by hydrolysis of alpha-tricalcium phosphate. The Ca-def HAp had a whisker-like morphology 2-5 microm in length and 0.1 microm in diameter that was elongated along c-axis. Thicker planer defects parallel to the (100) plane of the HAp matrix were observed as precipitation in the sample annealed at 700 and 800 degrees C by HRTEM observation. Thickness of the precipitation was about 10 nm and the boundaries between the precipitation and HAp matrix was coincident. The periodicity in the precipitation was parallel to the (100) plane of the HAp matrix and measured to be 1.42 nm. Since the precipitation was observed only in the sample annealed at a narrow temperature range of 700-800 degrees C, it was regarded as a metastable phase formed on the thermal decomposition process. Absorption peaks in IR spectra of annealed Ca-def HAp containing the metastable phase appeared at 744 and 3538 cm(-1) due to non-stoichiometric HAp with high Ca/P molar ratio. Furthermore, the results of energy dispersive X-ray spectroscopy showed that the metastable phase had higher Ca/P molar ratio than that of the matrix and stoichiometric HAp. Therefore, the metastable phase could be identified as Ca-rich metastable phase. The presence of Ca-rich metastable phase was confirmed to be associated with the thermal decomposition process. PMID- 15348425 TI - Porous phosphate-gelatine composite as bone graft with drug delivery function. AB - The design and synthesis of porous phosphate-gelatine composite implant which mimicks the structure of natural bone and has drug delivery function is proposed. Gelatine reproducing the proteinaceous part of bone was cross-linked in order to modulate its solubility in the physiologic fluids. The kinetic of gelatine release from ceramic matrix was also evaluated as model of the release of any therapeutic compound which can be loaded into gelatine. PMID- 15348426 TI - Characterization of hydrothermally treated anodic oxides containing Ca and P on titanium. AB - In this study, the formation of hydroxyapatite (HA) by a series of hydrothermal treatment and properties of the anodic oxide films and HA composite coatings were investigated. Based on the observations using EPMA, SEM, and X-ray diffraction, these needles were suggested to have a HA-like structure and were formed on the anodic oxide surfaces. The formation of these apatite-like needles were either oriented at an angle or formed on porous anodic oxide films. The concentration of Ca and P were enriched in apatite-like needles after hydrothermal treatments, whereas lower concentration of Ca and P were observed on anodic oxide film, particularly around the needles. In conclusion, HA-like needles were formed after hydrothermal treatment at 200 and 300 degrees C. PMID- 15348427 TI - Use of triethylcitrate plasticizer in the production of poly-L-lactic acid implants with different degradation times. AB - Bioabsorbable materials have been widely used in the repair of damaged tissue as well as in the controlled release of drugs and as a supports for cultured cells. The degradation time of poly-L-(lactic acid) (PLLA) may be controlled by altering the polymer porosity through the addition of the plasticizer triethylcitrate. This in turn influences the extent cellular infiltration. In this study, we examined the degradation of PLLA membranes containing different concentrations of plasticizer. PLLA discs were implanted subcutaneouly in rats and withdrawn 2, 14 and 60 days after implantation. The samples were processed for light microscopy and scanning electron microscopy (SEM). Polymer degradation was proportional to the concentration of plasticizer, indicating that triethylcitrate could affect the degradation time of the implants, without damaging the polymer biocompatibility. PMID- 15348428 TI - Synthesis and biocompatibility of porous nano-hydroxyapatite/collagen/alginate composite. AB - Porous nano-hydroxyapatite/collagen/alginate (nHAC/Alginate) composite containing nHAC and Ca-crosslinked alginate is synthesized biomimetically. This composite shows a significant improvement in mechanical properties over nHAC material. Mechanical test results show that the compressive modulus and yield strength of this composite are in direct proportion to the percentage of Ca-crosslinked alginate in the composite. Primary biocompatibility experiments in vitro including fibroblasts and osteoblasts co-culture with nHAC/alginate composite indicated the high biocompatibility of this composite. Therefore the composite can be a promising candidate of scaffold material for bone tissue engineering. PMID- 15348429 TI - Notch strength insensitivity of self-setting hydroxyapatite bone cements. AB - The effect of notches on the strength properties of self-setting hydroxyapatite (HA) cements is examined. Such stress concentrators may be present at orthopedic repair sites employing cements and significantly affect their mechanical reliability. Notched tensile specimens were prepared from two cement compositions that resulted in HA and carbonated apatite. The notch radii was varied from 0.15 to 6 mm with a fixed length of 6 mm. The strength of the cements was found to be surprisingly insensitive to the presence of the notches over the range of notch radii examined. A fracture statistics model incorporating a Weibull statistical approach was employed to rationalize the observed notch insensitivity. PMID- 15348430 TI - Synthesis and characterization of nano-HA/PA66 composites. AB - Based on the bioactivity and biocompatibility of hydroxyapatite (HA) and the excellent mechanical performance of polyamide 66 (PA66), a composite of nanograde HA with PA66 was designed and fabricated to mimic the structure of biological bone which exhibits a composite of nanograde apatite crystals and natural polymer. The HA/PA66 composite combines the bioactivity of HA and the mechanical property of PA66. This study focused on the preparation method of HA/PA66 composite and the influence of HA crystals on the characterization of the composite. HA slurry was used directly to prepare HA/PA66 composite by a solution method, in which HA is able to form hydrogen bond, i.e. chemical bonding with PA66. The nano-HA needle-like crystals treated by hydrothermal method are better in the particle size distribution and the particle dispersion. The morphology, crystal structure and crystallinity as well as crystal size of these needle-like crystals are similar to bone apatite. The nano-HA needle-like crystals dispersed uniformly in PA66 matrix with reinforcement effect and can prevent the micro crackle spreading into cleft and fracture during the deformation process. The mechanical testing shows that the nano-HA/PA66 composite has a good mechanical property, and may be a promising bone replacement material. PMID- 15348431 TI - Coupling of HDPE/hydroxyapatite composites by silane-based methodologies. AB - Several coupling treatments based on silane chemicals were investigated for the development of high density (HDPE)/hydroxyapatite (HA) composites. Two HA powders, sintered HA (HAs) and non sintered HA (HAns), were studied in combination with five silanes, namely y-methacryloxy propyltrimethoxy silane (MEMO), 3-(2-aminoethyl)aminopropyltrimethoxy silane (DAMO), vinyltrimethoxy silane (VTMO), 3-aminopropyltriethoxy silane (AMEO) and trimethoxypropyl silane (PTMO). The HA particles were treated by a dipping in method or by spraying with silane solutions. After drying, the treated powders were compounded with HDPE or HDPE with acrylic acid and/or organic peroxide and subsequently compression molded. The tensile test specimens obtained from the molded plates were tensile tested and their fracture surfaces were observed by scanning electron microscopy (SEM). For the sintered HA (HAs) composites, the most effective coupling treatments concerning stiffness are those based on MEMO and AMEO. The low influence of these coupling procedures on strength is believed to be associated to the low volume fraction and the relatively smooth surface of the used HA particles. For the non-sintered HA (HAns) composites, it was possible to improve significantly both the stiffness and the strength. Amino silanes demonstrated to be highly efficient concerning strength enhancement. The higher effectiveness of the coupling treatments for HAns filled composites is attributed to their higher particle surface area, smaller particle size distribution and expected higher chemical reactivity. For both cases, the improvement in mechanical performance after the coupling treatment is consistent with the enhancement in interfacial adhesion observed by SEM. PMID- 15348432 TI - Apatite-forming ability of glass-ceramic apatite-wollastonite - polyethylene composites: effect of filler content. AB - The bioactivity of a range of glass-ceramic apatite-wollastonite (A-W) - polyethylene composites (AWPEXs) with glass-ceramic A-W volume percentages ranging from 10 to 50, has been investigated in an acellular simulated body fluid (SBF) with ion concentrations similar to those of human blood plasma. The formation of a biologically active apatite layer on the composite surface after immersion in SBF was demonstrated by thin-film X-ray diffraction (TF-XRD) and field-emission scanning electron microscopy (FE-SEM). An apatite layer was formed on all the composites, with the rate of formation increasing with an increase in glass-ceramic A-W percentage. For composites with glass-ceramic A-W filler contents >or=30 vol %, the apatite layer was formed within 12 h of immersion, which is a comparable time for apatite formation on monolithic glass-ceramic A-W. Inductively coupled plasma atomic emission spectroscopy (ICP-AES) demonstrated that the apatite formation on AWPEX samples with 50 vol % filler content occurred in a manner similar to that seen on pure glass-ceramic A-W, in that the calcium, silicon, and magnesium ion concentrations increased and, conversely, a decrease was observed in the phosphate ion concentration. These results indicate that a suitable in vitro response was achieved on a composite incorporating particulate glass-ceramic A-W with a particularly favorable response being observed on the AWPEX sample with 50 vol % filler content. PMID- 15348433 TI - The effect of resin/crosslinker ratio on the mechanical properties and fungal deterioration of a maxillofacial silicone elastomer. AB - Variation of the crosslinker/resin ratio of a room temperature condensation cure maxillofacial silicone elastomer has caused considerable changes in the mechanical properties and deterioration by Candida albicans. Increasing the crosslinker/resin ratio caused a decrease in the tensile strength and stiffness of the elastomer. However, tear strength appeared to show an optimum value at the recommended crosslinker/resin ratio. These effects were due to the low molar mass silicone polymer that acts as a carrier for the actual crosslinking additive. The general decrease in mechanical properties was accompanied by an increase in the hexane extractables content and an increase in the Si-H content of the elastomer. The unbound polymer (extractable material) content of the elastomer was found to influence the colonization of the material by C. albicans. An increase in the unbound polymer content corresponded to an increasing number of hyphae and blastospores observed penetrating into the elastomer. The data obtained in this study have significant implications concerning the degree of control of elastomer formulation and the deterioration of maxillofacial appliances. PMID- 15348434 TI - Calcium phosphate nucleation on surface-modified PTFE membranes. AB - Highly porous PTFE membranes are currently being used in facial reconstructive surgery. The present study aims at improving this biomaterial through creating a more bioactive surface by introducing ionic groups onto the surface. The unmodified PTFE membrane does not induce inorganic growth after immersion in simulated body fluid (SBF) for up to 4 weeks. Copolymeric grafting with acrylic acid (AAc) by means of gamma irradiation and subsequent in vitro testing in SBF reveals that this copolymer initially acts as an ion-exchange material and subsequently induces growth of a calcium phosphate phase (Ca/P=2.7) when large amounts (15%) of pAAc are introduced onto the membrane surface. This copolymer is not expected to function well from a biomaterials perspective since SEM showed the pores on the surface to be partly blocked. In contrast, the surface of monoacryloxyethyl phosphate (MAEP)-modified samples is altered at a molecular level only. Yet the modified materials are able to induce calcium phosphate nucleation when the external surface coverage is 44% or above. The initial inorganic growth on these membranes in SBF has a (Ca+Mg)/P ratio of 1.1 (presumably Brushite or Monetite). The secondary growth, possibly calcium deficient apatite or tricalcium phosphate, has a (Ca+Mg)/P ratio of 1.5. This result is a promising indicator of a bioactive biomaterial. PMID- 15348435 TI - In vitro cellular response to titanium electrochemically coated with hydroxyapatite compared to titanium with three different levels of surface roughness. AB - The in vitro response of primary human osteoblast-like (HOB) cells to a novel hydroxyapatite (HA) coated titanium substrate, produced by a low temperature electrochemical method, was compared to three different titanium surfaces: as machined, Al(2)O(3)-blasted, plasma-sprayed with titanium particles. HOB cells were cultured on different surfaces for 3, 7 and 14 days at 37 degrees C. The cell morphology was assessed using scanning electron microscopy (SEM). Cell growth and proliferation were assessed by the measurement of total cellular DNA and tritiated thymidine incorporation. Measurement of alkaline phosphatase (ALP) production was used as an indicator of the phenotype of the cultured HOB cells. After three days incubation, the electrochemically coated HA surface produced the highest level of cell proliferation, and the Al(2)O(3)-blasted surface the lowest. Interestingly, as the incubation time was increased to 7 days all surfaces produced a large drop in tritiated thymidine incorporation apart from the Al(2)O(3)-blasted surface, which showed a small increase. Cells cultured on all four surfaces showed an increased expression of ALP with increased incubation time, although there was not a statistically significant difference between surfaces at each time point. Typical osteoblast morphology was observed for cells cultured on all samples. The HA coated sample showed evidence of a deposited phase after three days of incubation, which was not observed on any other surface. Cells incubated on the HA coated substrate appeared to exhibit the highest number of cell processes attaching to the surface, which was indicative of optimal cell attachment. The crystalline HA coating, produced by a low temperature route, appeared to result in a more bioactive surface on the c.p. Ti substrate than was observed for the other three different Ti surfaces. PMID- 15348436 TI - Effect of annealing titanium on in vitro bioactivity. AB - In order to modify titanium surfaces for various biological applications, bioactive and pure titanium oxide thin films were coated on the titanium by thermal oxidation technique. The commercially pure titanium discs after polishing were heated at 500, 550, 600, 650 and 700 degrees C, respectively, for 10 min in air or in argon. To evaluate the ability of calcium phosphate formation, samples after annealing were soaked in the Eagle's minimum essential medium solution. Surface morphology and chemical composition of the samples before or after immersion were characterized by field emission - scanning electron microscopy and energy dispersive X-ray spectrometry. PMID- 15348437 TI - Biological and biochemical properties of the carbon composite and polyethylene implant materials. AB - We studied the biocompatibility of the carbon composites and polyethylene materials with and without collagen or collagen and proteoglycan cover. We used the in vitro technology to study the adhesion of model cells evalution, their metabolic activity and the production of TNF-alpha as a cytokine model. Under in vivo condition, the biocompatibility of tested polymers were studied in the implantation experiment, subcutaneously in the interscapular region in the laboratory rat. We have found in the in vitro assay favorable proliferation and the smallest production of pro-inflammatory TNF-alpha cytokine in cells adherent to the hydrophobic polyethylene material coated with biological macromolecules. Using in vivo tests performed by the implantation of materials to the rat we demonstrated that the materials are not cytotoxic. The tissue capsule surrounding the implants was not significantly influenced by the type of the implant and the pre-treatment by the biological molecules. However, the foreign-body giant multinucleated cells were observed only in the vicinity of the collagen - covered hydrophobic polyethylene implant. Interestingly, while the collagen coating improved the biocompatibility of tested polymers in vitro, the inflammatory reaction against this covered materials was higher under in vivo conditions. The pre-treatment of carbon composites by both types of biological macromolecules reduced the occurrence of carbon debris in the implantation site. The tested carbon composites and polyethylene materials are not toxic. The pre-treatment of the materials by extracellular matrix components increased their biological tolerance in vitro and reduced implant wears in animal experiment, which can be important for the medical application. PMID- 15348438 TI - Biological behavior of sol-gel coated dental implants. AB - The biocompatibility of dental implants coated with titania/hydroxyapatite (HA) and titania/bioactive glass (BG) composites obtained via sol-gel process was investigated using an in vitro and in vivo model. A device for the in vitro testing of screw-shaped dental implants was developed, in order to well compare the two experimental models studying the behavior of human MG63 osteoblast-like cells seeded onto a particular geometry. The expression of some biochemical parameters of osteoblastic phenotype (alkaline phosphatase specific activity, collagen and osteocalcin production) and some indications on cells morphology obtained by scanning electron microscopy were evaluated. The in vitro and in vivo models were compared after implants insertion in rabbit tibia and femur. The removal torque and histomorphometric parameters (percentage of bone in contact with implant surface and the amount of bone inside the threaded area) were examined. A good agreement was found between the in vitro and in vivo models. These experiments showed better performances of HA and BG sol-gel coated dental implants with respect to uncoated titanium; in particular, it was found that in vitro the HA coating stimulates osteoblastic cells in producing higher level of ALP and collagen, whereas in vivo this surface modification resulted in a higher removal torque and a larger bone-implant contact area. This behavior could be ascribed to the morphology and the chemical composition of the implants with rough and bioactive surfaces. PMID- 15348439 TI - Strength of bond to bone and cytotoxicity of sintered bodies of hydroxyapatite/zirconia composite particles. AB - Regarding sintered bodies of hydroxyapatite (HAP)/zirconia (PSZ) composite particles prepared by covering the surface of HAP particles with PSZ particles, their strength of the bond to bone after implanting them into an organism and their cytotoxicity were evaluated. Cytotoxicity tests were conducted by the colony formation method. Cytotoxicity was not observed in the sintered bodies of HAP/PSZ composite particles, or in sintered monolithic HAP body and sintered monolithic PSZ body. The strength of bond between a sintered body and a bone was evaluated by measuring the shear strength at the interface between them after a fixed period following implantation of a sintered body into a rabbit femur. In all cases of the sintered monolithic HAP body, and the sintered bodies of HAP/PSZ composite particles and monolithic PSZ body, when the implantation period increased, the shear strength of the sintered body/bone interface tended to increase. In particular, this tendency was very high in the case of sintered bodies of HAP/PSZ with weight ratios of 1/1.0 and 1/1.5, the shear strength for each reaching 20 MPa 12 months after implantation. PMID- 15348440 TI - On the surface elemental composition of non-corroded and corroded dental ceramic materials in vitro. AB - Dental ceramics are traditionally looked upon as inert materials. As many are glass phased, it may be hypothesized that they will be subjected to glass corrosion in aqueous environments. The aim of the study was therefore to analyze the surface elemental composition of glass-phased and all-crystalline ceramics, before and after low- and high-intensity, in vitro corrosion (milli-Q-water at 37+/-2 degrees C for 18 h and 4% acetic acid at 80+/-2 degrees C for 18 h, respectively). The analysis of the surface elemental composition was performed using ESCA. The hypothesis was confirmed. After high-intensity corrosion, the complete wash out of alkali ions, alkaline-earth ions and elemental alumina was found, leaving behind a surface totally dominated by silica. The all-crystalline ceramics, densely sintered alumina and yttria-partially stabilized tetragonal zirconia, displayed only minor surface changes, even after high-intensity corrosion. In comparison to the corrosion testing in acid, the corrosion process in milli-Q-water did not produce different results in principle, except for the lower magnitude of the depletion of alkali ions and the virtually unchanged level of elemental alumina. Unexpectedly, no substantial difference in surface degradation was found between the glass ceramic and the ordinary porcelain-fused to-metal ceramic or between ceramics of higher sintering temperature and those of low or ultra-low sintering temperature. The composition and microstructure alone did not appear to provide a full explanation for the inter-individual differences in surface corrosion when exposed to comparable environmental conditions. PMID- 15348441 TI - Bi-composite sandwich moldings: processing, mechanical performance and bioactive behavior. AB - Two composite systems composed of high-density polyethylene (HDPE) filled with hydroxyapatite (HA) and carbon fiber (C fiber) were compounded in a co-rotating twin screw extruder and subsequently molded in a two component injection molding machine in order to produce test bars with a sandwich-like morphology. These moldings are based on a HDPE/HA composite outer layer and a HDPE/C fiber composite core. The mechanical performance of the obtained specimens was assessed by tensile and impact testing. The fracture surfaces were observed by scanning electron microscopy (SEM) and optical reflectance microscopy was used to characterize the morphology within the moldings. In order to study the bioactivity of the molded specimens, the samples were immersed for different periods of time up to 30 days in a simulated-body fluid (SBF) with an ion composition similar to human blood plasma. After each immersion period, the surfaces of the specimens were characterized by SEM. The chemical composition and the structure of the deposited films were studied by electron dispersive spectroscopy (EDS) and thin-film X-ray diffraction (TF-XRD). The evolution of the elemental concentrations in the SBF solution was determined by induced coupled plasma emission (ICP) spectroscopy. Bi-composite moldings featuring a sandwich like morphology were successfully produced. These moldings present a high stiffness as a result of the C fiber reinforcement present in the molding core. Furthermore, as a result of the HA loading, the sandwich moldings exhibit a clear in vitro bioactive behavior under simulated physiological conditions, which indicates that an in vivo bone-bonding behavior can be expected for these materials. PMID- 15348442 TI - Factors influencing the compressive strength of an injectable calcium sulfate hydroxyapatite cement. AB - A biphasic injectable bone substitute, suitable for filling bone defects, that sets in the body, based on calcium sulfate and hydroxyapatite (HA), is presented. For applications in bone defects the compressive strength is important to assure support of the defect site during loading when the patient is weight bearing. To control the strength, the influence of four different factors; the liquid-to powder (L/P) ratio, the HA particle morphology, the HA content and the amount of accelerator, were investigated. alpha-Calcium sulfate hemihydrate (CSH) and four different HA powders (three sintered and one spray-dried) were used. All differed in size and morphology. CSH and each HA powder were mixed together with distilled water to form the bone substitute. An accelerator, in form of calcium sulfate dihydrate, was added to the powder phase to obtain an adequate setting time. Cylindrical specimens were compression tested. A lower L/P-ratio gave stronger cement, but was more difficult to inject. The shape and the morphology of the HA particles influenced the strength, and reducing the amount of HA increased the strength. The amount of accelerator (calcium sulfate dihydrate) had no influence. PMID- 15348443 TI - Effect of surface topography on the color of dental porcelain. AB - The objectives of this study were to evaluate the color difference depending on the surface topography of roughness and glazing, and to determine the effects of color measuring geometry and the standard illuminant on the color of a dental porcelain. Disk specimens of A3 shade were fired with commercial dental porcelain for PFM. Specimens were divided into non-polished (ST 1), polished with 200, 400, 1000, 1500-grit SiC papers (ST 2, 3, 4, 5) and glazed (ST 6) groups. After measuring the average surface roughness (Ra), color was determined under the illuminant A and D65 on a spectrophotometer with the specular component excluded (SCE) and included (SCI) geometry. Ra values were significantly influenced by the surface topography. With the SCE, the CIE L* value after glazing was significantly lower than that after polishing. Color differences (DeltaE*) measured with the SCE were higher than those with the SCI (2.61-4.66 vs. 0.93 1.57). Therefore the SCE geometry seemed to more accurate protocol for the color measurement of dental porcelain. PMID- 15348444 TI - Sutured calf pericardium: influence of the type and angle of the suture on mechanical behavior. AB - Careful selection of the biological tissue to be used in cardiac bioprostheses and a thorough knowledge of its mechanical behavior, facilitating both the prediction of this behavior and the interactions between the tissue and the other materials employed, is the best approach to designing a durable implant. For this purpose, a study involving uniaxial tensile testing of calf pericardium was carried out. Two sets of three contiguous strips of tissue were obtained from each pericardial membrane, to perform a total of 144 trials. Two samples were sewn with one of four commercially available suture materials: Gore-Tex, nylon, Prolene and silk. In each set of three samples, the center strip remained intact and unsutured to serve as a control, while the left-hand strip was sutured at a 45 degrees angle with respect to the longitudinal axis and the right-hand strip was sewn at a 90 degrees angle. All the samples were tested until rupture. The results demonstrated a significant loss of mean load (p<0.01) in the sutured samples at rupture. The angle of the suture had no influence on these results, although the stress/strain curves showed that, as the tensile stress increased, the mechanical behaviors were less uniform. The rupture of the collagen fibers could explain this phenomenon. The pericardium sutured with Gore-Tex presented a greater strain, or deformation (elongation), at lower levels of stress, regardless of the angle of the suture. The tissue selection criteria, based on the use of paired samples, enabled a correct prediction of the mechanical behavior of the tissue, with excellent correlation coefficients (>0.98) and a high degree of homogeneity in the results. PMID- 15348445 TI - Calcium phosphate interactions with titanium oxide and alumina substrates: an XPS study. AB - Besides the excellent mechanical properties of titanium and alumina (Al(2)O(3)) in the case of load bearing applications, their bone-bonding properties are very different. In osseous environment, Al(2)O(3) ceramic is encapsulated by fibrous tissues, whereas bone can bind directly to titanium, via its natural titanium dioxide (TiO(2)) passivation layer. So far, this calcification dissimilarity between TiO(2) and Al(2)O(3) was attributed to respectively their negative and positive surface charge under physiological conditions. The present study aims at studying the chemical interactions between TiO(2) and Al(2)O(3) (phase alpha) with the diverse ions contained in simulated body fluids (SBFs) buffered with trishydroxymethyl aminomethane (TRIS) at pH=6.0 and pH=7.4. After 1 h of immersion, TiO(2) and alpha-Al(2)O(3) powders were analyzed by X-ray photoelectron spectroscopy (XPS). The results indicated that Ca and HPO(4) groups were present on TiO(2) surface. In addition, HPO(4) groups were found to be in a higher amount than Ca on TiO(2), which does not comply with the surface charge theory. With regard to Al(2)O(3), little HPO(4) but no Ca was detected on its surface, and TRIS bound to Al(2)O(3) substrate in all of the immersion experiments. The fact that both Ca and HPO(4) were present at the vicinity of TiO(2) might be at the origin of its calcification ability. On the other hand, Al(2)O(3) did not show any affinity towards Ca and HPO(4) ions. This might explain the inability of Al(2)O(3) substrate to calcify. PMID- 15348446 TI - Microstructure and anodic polarization behavior of experimental Ag-18Cu-15Pd-12Au alloy in aqueous sulfide solution. AB - The anodic corrosion behavior of an experimental Ag-15Pd-18Cu-12Au alloy in 0.1% Na(2)S solution in relation to its microstructure was investigated using potentiodynamic and potentiostatic polarization techniques with analyses of corrosion products by X-ray diffractometry, Auger electron spectroscopy, and X ray photoelectron spectroscopy. The role of Pd in improvement of the corrosion resistance was also investigated. In the potential/current density curve, three distinct current peaks, at -520 mV (peak I), -425 mV (peak II) and -175 mV (peak III), were observed. The Ag-rich alpha(2) matrix with coarse Cu and Pd-rich lamellae was the most corrosion-susceptible region, and this region was preferentially corroded at peak I with the formation of granular deposits of Ag(2)S. A small amount of Ag-Cu mixed sulfide deposited on the Cu and Pd-rich coarse particles and dissolution of Ag as AgO(-) might have occurred in parallel with Ag(2)S formation at peak II. Enrichment of Pd on the alloy surface occurred at peak III due to preferential dissolution of Ag and Cu. A high level of corrosion resistance was attained with the formation of a thin Pd-rich sulfide film, which enhanced the passivity of the alloy in an alkaline sulfide solution. It was found that passivity is an important phenomenon not only for base metal alloys but also for noble metal alloys to maintain high levels of resistance to corrosion and tarnishing in sulfide environments. PMID- 15348447 TI - An innovative auto-catalytic deposition route to produce calcium-phosphate coatings on polymeric biomaterials. AB - The aim of this research is to develop a new methodology to obtain bioactive coatings on bioinert and biodegradable polymers that are not intrinsically bioactive. In this study three types of materials were used as substrates: (i) high molecular weight polyethylene (HMWPE) and two different types of starch based blends (ii) starch/ethylene vinyl alcohol blends, SEVA-C and (iii) starch/cellulose acetate blends, SCA. Two types of baths were originally proposed and studied to produce novel auto-catalytic calcium-phosphate (Ca-P) coatings. Then, the coated surfaces were analyzed by scanning electron microscopy and energy dispersive spectroscopy (SEM/EDS), as produced, and after different immersion periods in SBF. The evolution of Ca and P concentrations was determined by induced-coupled plasma emission (ICP) spectroscopy. The crystalline phases present on the films formed on the different material surfaces, after a certain soaking time, were identified by thin-film X-ray diffraction (TF-XRD). The obtained results indicated that it was possible to coat the materials surfaces with a Ca-P layer with only 60 min of immersion in both types of auto-catalytic solutions. Furthermore, it was possible to observe the clear bioactive nature of the Ca-P coatings after different immersion periods in a simulated body fluid (SBF). The results from TF-XRD confirmed the presence of partially amorphous Ca-P films with clearly noticeable hydroxylapatite peaks. These new methodologies allow for the production of an adherent bioactive film on the polymeric surfaces prior to implantation, which may allow for the development of bone-bonding, bioabsorbable implants and fixation devices. PMID- 15348448 TI - Bioresorbable and bioactive composite materials based on polylactide foams filled with and coated by Bioglass particles for tissue engineering applications. AB - Poly(DL-lactide) (PDLLA) foams and bioactive glass (Bioglass) particles were used to form bioresorbable and bioactive composite scaffolds for applications in bone tissue engineering. A thermally induced phase separation process was applied to prepare highly porous PDLLA foams filled with 10 wt % Bioglass particles. Stable and homogeneous layers of Bioglass particles on the surface of the PDLLA/Bioglass composite foams as well as infiltration of Bioglass particles throughout the porous network were achieved using a slurry-dipping technique. The quality of the bioactive glass coatings was reproducible in terms of thickness and microstructure. In vitro studies in simulated body fluid (SBF) were performed to study the formation of hydroxyapatite (HA) on the surface of the PDLLA/Bioglass composites, as an indication of the bioactivity of the materials. Formation of the HA layer after immersion in SBF was confirmed by X-ray diffraction and Raman spectroscopy measurements. The rate of HA formation in Bioglass-coated samples was higher than that observed in non-coated samples. SEM analysis showed that the HA layer thickness rapidly increased with increasing time in SBF in the Bioglass coated samples. The high bioactivity of the developed composites suggests that the materials are attractive for use as bioactive, resorbable scaffolds in bone tissue engineering. PMID- 15348449 TI - Thermoresponsive hydrogel with rapid response dynamics. AB - Intelligent hydrogels, particularly poly(N-isopropylacrylamide) (PNIPAAm)-based hydrogels, have attracted extensive interest because the soft wet hydrogels can change their shapes in response to the small changes of environmental factors like temperature. In order to fully make use of this unique property of PNIPAAm based hydrogels, the response rates of the PNIPAAm hydrogels have to be improved since the dynamics property is critical to certain applications of this material. In this paper, the thermo-sensitive PNIPAAm hydrogels were successfully synthesized by carrying out the polymerization of N-isopropylacrylamide monomer in vacuum (-100 kPa) at room temperature (22 degrees C). The resultant hydrogel has tremendously improved shrinking rate as well as the large volume changes upon temperature stimulation when comparing with the normal PNIPAAm hydrogel. The SEM micrographs revealed that the improved properties were attributed to the macroporous network structure generated during the synthesis under vacuum. PMID- 15348450 TI - The distribution of water in degrading polyglycolide. Part I: sample size and drug release. AB - The effect of sample thickness on the degradation of polyglycolide (PGA) disks and on their drug release profiles is explored in this paper, and conclusions drawn about the distribution of water across a sample during degradation. The degradation process was monitored by measuring changes in the long period calculated from small angle X-ray scattering profiles, and by following changes in the pH of the buffer solutions. Drug release profiles were obtained using UV spectrophotometry. The measurements suggest that reaction-erosion fronts form at the surface of all samples after around 7 days of degradation, and that these fronts progress through the sample at a constant rate of 0.032 mm/day. The data are consistent with a model in which drug is released quickly from the porous, hydrated regions behind the front, and reaches 100% release when the fronts meet. PMID- 15348451 TI - The distribution of water in degrading polyglycolide. Part II: magnetic resonance imaging and drug release. AB - This paper reports the use of magnetic resonance imaging (MRI) on polyglycolide disks to monitor the change in water ingress with degradation time. Very little response was measured before 13 days, but after this time, water began to penetrate the disks as fronts, starting from the sample surface and moving inwards towards the centre. These results provide more direct evidence in support of the four-stage degradation model for PGA outlined in previous literature, and in particular, that fairly sharp reaction-erosion fronts move in from the sample surface to the centre when the polymer is undergoing significant mass loss and water gain. A combination of MRI and drug release data suggest that fronts originate at the surface at about 7 (+/-2) days, and proceed at a rate of 0.033 (+/-0.002) mm/day. These results agree with results obtained from cumulative drug release profiles for different sample thicknesses presented in Part I. They support the hypothesis that drug releases quickly from the swollen regions behind the fronts where the polymer is open and porous, and that release finishes when the fronts meet in the centre of the sample. PMID- 15348452 TI - Torsional fatigue resistance of plasma sprayed HA coating on Ti-6Al-4V. AB - The torsional strength of plasma sprayed hydroxyapatite (HA) coatings was studied under static and cyclic loading. The torsional shear tests were conducted in a frustum test device developed in this laboratory, which adapted to various coating thicknesses. The interfacial fatigue resistance was measured in terms of interfacial fatigue strength defined as the average maximum stress (tau(fmax)). A staircase fatigue method was employed to determine the interfacial fatigue strength; this method resolved the uncertainty in detecting coating failure during torsion fatigue. The values for coating shear strength and shear fatigue strength obtained from the torsional tests did not differ from those obtained by previous tensional shear tests in this laboratory. The fatigue strength of one million cycles was about 35% lower than static shear strength. This finding might be used for estimating fatigue life span without cyclic loading tests. PMID- 15348453 TI - Primary bone-derived cell colonization of unconditioned and pre-conditioned Bioglass 45S5 surfaces in vitro. AB - Samples of Bioglass 45S5, a bioactive glass that reacts chemically on immersion in an aqueous environment and following implantation can bond with bone, were immersed in culture medium with foetal calf serum as a source of proteins for periods of 1-11 days. Energy dispersive spectroscopy (EDS) revealed that over 11 days immersion, the Bioglass surface was rich in Si relative to Ca and P and scanning electron microscopy (SEM) showed that the Bioglass developed surface reaction layers. Samples conditioned in culture medium and previously untreated samples were used as substrates for primary osteoblast or periosteal cell cultures, to allow comparison of the effects of different Bioglass surface development, on subsequent cell attachment. SEM examination revealed that pre conditioned Bioglass was a more suitable substrate for osteoblast colonization than previously undeveloped Bioglass, whereas periosteal cells colonized all samples equally well and more rapidly than osteoblasts. PMID- 15348454 TI - Effects of dental resin metabolites on estrogenic activity in vitro. AB - Three monomers (Bis-GMA, UDMA, and TEGDMA) and five polymerization initiators (CQ, BPO, DMPT, DMAEMA, and ATU) commonly used in dental composite resins were tested for estrogenic activity using a reporter gene assay (yeast two-hybrid system) in vitro, and compared with bisphenol-A (BPA). Estrogenic activity was indicated by agonist and antagonist activity, with (+S9) and without (-S9) metabolic activation using rat liver cells. No estrogenic agonist activity was seen for each monomer and polymerization initiator in either the -S9 and +S9 tests in the concentration ranges examined in this study. On the other hand, estrogen antagonist activity was found with BPO and DMPT. BPO showed antagonist activity at a concentration of approximately 1800 nM with the -S9 test, but not with the +S9 test. With DMPT, antagonist activity was not seen with the -S9 test, but it was seen at a concentration of approximately 610 nM using the +S9 test. With BPA, the +S9 test indicated antagonist activity at a concentration of approximately 780 nM. The estrogen antagonist activities of DMPT and BPA appeared to be similar. CQ, DMAEMA, ATU, and the three monomers did not show antagonist activity as demonstrated by the -S9 or +S9 tests within the concentration range tested in this study. PMID- 15348455 TI - Synthesis and characterization of hydroxyapatite, fluoride-substituted hydroxyapatite and fluorapatite. AB - Powders of hydroxyapatite (HA), partially fluoride-substituted hydroxyapatite (fHA), and fluorapatite (FA) were synthesized in house using optimum methods to achieve relatively pure powders. These powders were assessed by the commonly used bulk techniques of X-ray diffraction (XRD), Fourier transform infra-red (FTIR) and FT-Raman spectroscopies, inductively coupled plasma atomic emission spectroscopy (ICP-AES), and F-selective electrode. In addition, the current study has employed transmission electron microscopy (TEM), involving morphological observation, electron diffraction and energy-dispersive X-ray spectrometry (EDX), as an effective analytical technique to evaluate the powders at a microscopic level. The HA and fHA particles were elongated platelets about 20 x 60 nm in size, while FA particles were over twice this size. Calcination of the HA and fHA powders at 1000 degrees C for 1 h resulted in increased grain size and crystallinity. The calcined fHA material appeared to possess a crystal structure intermediate between HA and FA, as evidenced by the (3 0 0) peak shift in XRD, as well as by the position of the hydroxyl bands in the FTIR spectra. This result was consistent with electron diffraction of individual particles. Small levels of impurities in some of the powders were identified by EDX and electron diffraction, and the carbonate content was detected by FTIR. The use of TEM in conjunction with the bulk techniques has allowed a more thorough assessment of the apatites, and has enabled the constituents in these closely related apatite powders to be identified. PMID- 15348456 TI - Creep behavior of bone cement: a method for time extrapolation using time temperature equivalence. AB - The clinical lifetime of poly(methyl methacrylate) (PMMA) bone cement is considerably longer than the time over which it is convenient to perform creep testing. Consequently, it is desirable to be able to predict the long term creep behavior of bone cement from the results of short term testing. A simple method is described for prediction of long term creep using the principle of time temperature equivalence in polymers. The use of the method is illustrated using a commercial acrylic bone cement. A creep strain of approximately 0.6% is predicted after 400 days under a constant flexural stress of 2 MPa. The temperature range and stress levels over which it is appropriate to perform testing are described. Finally, the effects of physical aging on the accuracy of the method are discussed and creep data from aged cement are reported. PMID- 15348457 TI - Histomorphological study of bone response to hydroxyapatite coating on stainless steel. AB - Bone response to hydroxyapatite coating on stainless steel has not been so extensively tested in animals as it happened for other metallic substrate, like Ti6Al4V. For this reason, authors performed an in vivo histomorphological electron microscopic study of hydroxyapatite coating on duplex stainless steel cylinders, to gather further evidences on the characters of bone apposition at the interface. Sixteen HA-coated cylinders were implanted in the distal femur of New Zealand White rabbits. Comparison with uncoated controls was made. Retrieval steps were at: 4, 8, 26 and 34 weeks. Specimens were analyzed in a Jeol JSM 6301F scanning electron microscope. The response to HA-coated samples has a morphological character of tight apposition between bone and coating. Osteocytic lacunae may be found few microns close to the coating and newly formed bone is extremely interlocked with it so that even an higher magnification electron microscopy cannot resolve any discontinuity in between. Pictures of physiological bone-turnover are distinguishable at the bone-coating interface; areas of well preserved coating may be present together with areas where local exfoliation or fragmentation has already completely exposed the metallic substrate. On the opposite in uncoated samples, despite a morphological picture of properly formed bone, the largest area of the metal has no direct apposition with it. PMID- 15348458 TI - Testing the cytotoxicity of metal alloys used as magnetic prosthetic devices. AB - Technical magnetic materials are increasingly used for the development of magnetic retained dental prosthetic and orofacial epithetic devices. Since most of the magnets based on rare earth metals, such as samarium-cobalt based alloys have a high tendency for corrosion they were first coated by tin and then encapsulated by titanium. However, the high mechanical load particularly on dental devices may cause a rupture of the titanium capsule and the alloys contact directly biological fluids. Hence, it is important to know the cytotoxicity of these magnets to assess their potential effects on the surrounding tissue. In this study, the cytotoxicity of neodymium-iron-boron and samarium-cobalt (plain, tin and titanium coated) magnets was tested. First, magnets were incubated up to 7 days in culture medium to prepare extracts for cytotoxicity measurements. Changes in the surface morphology due to corrosion were visualized by scanning electron microscopy and analysis of the elemental composition. 3T3 mouse fibroblasts were cultured in the presence of extracts and their viability measured by neutral red and metabolic assays. To learn more about a possible toxic activity of the main components of magnets, salt solutions of different concentrations resembling those elements, which are main constituents of the magnets, were used. 3T3 fibroblasts were also cultured in direct contact with the materials and material induced effects on cell morphology and growth monitored by microscopy. As a result of this study it was found that samarium-cobalt magnets have a strong tendency for corrosion and exert a considerable cytotoxicity. Neodymium-iron-boron magnets have a lesser tendency for corrosion and are only moderate cytotoxic. Coating of samarium-cobalt magnets with tin or titanium makes the materials non-toxic. Application of salt solutions shows that cobalt has a tendency to be cytotoxic at higher concentrations, but enhances cell metabolism and proliferation at lower concentrations while the other magnet constituents had a lower or negligible cytotoxic potential. PMID- 15348459 TI - Release dynamics of tetracycline from a loaded semi-interpenetrating polymeric material of polyvinyl alcohol and poly(acrylamide-co-styrene). AB - A semi-interpenetrating polymer network (IPN) of polyvinyl alcohol (PVA) and poly(acrylamide-co-styrene) (PAMS) was prepared and its potential for controlled release of tetracycline was assessed. The IPN was characterized by IR spectral analysis and network parameters such as the average molecular weight between crosslinks (M(c)), crosslink density (q) and number of elastically effective chains (V(e)) were evaluated. The influence of various experimental conditions such as different percent loadings, composition of the IPNs, thickness of the loaded device, pH and nature of the release medium were investigated on the release profiles of the drug. Various kinetic constants such as the diffusional exponent (n), diffusion constant (D) and penetration velocity (v) were evaluated for different release processes and based on the dynamic release data, an analysis of transport mechanisms of tetracycline was made using Fick's equations. PMID- 15348460 TI - In vitro mesothelialization of prosthetic materials designed for the repair of abdominal wall defects. AB - The aim of this study was to evaluate the in vitro response of mesothelial cells (MC) in terms of their ability to cover different biomaterials. MC were harvested from human omentum. The MC from the first passage were seeded onto different biomaterials from 10 min to 24 h: PL-PU99 (polypropylene-polyurethane); DM (ePTFE); PL (polypropylene); and PL + Col (polypropylene-collagen). The prosthetic surface covered was examined by microscopy and quantified. PL-PU99: The MC were adhered to the biomaterial 10 min post-incubation. At 4 h, the 53.12+/-7.86% of the prosthesis were coated with polygonal cells. At 12 h, 96.32+/-11.32% of the biomaterial was coated. DM: between 30 min to 8 h, the MC cells form small, round colonies. At 12 h, polygonal and fusiform secretory cells were observed (68.94+/-5.78%). 93.54+/-11.49% of surface was coated after 24 h. PL: only isolated cells were observed on the prosthesis. PL + Col: MC form a monolayer over prosthetic surface after 18 h (90.21+/-9.76). We conclude: (a) MC formed a stable monolayer over all the biomaterials tested with the exception of the PL due to its porosity. (b) The PL-PU99 showed the greatest potential for in vitro mesothelialization compared to the PL-Col and DM prostheses. PMID- 15348461 TI - Granule size and composition of bioactive glasses affect osteoconduction in rabbit. AB - Bioactive glass granules of three different compositions, regarding particularly Si- and Al- content (S53P4, S59.7P2.5, S52P3) and of two different granule sizes (200-250 microm and 630-800 microm) were implanted for 4 and 8 weeks in the distal part of rabbit femur. The effect of glass composition and granule size on bone formation was studied. The results were evaluated using histology, computerized histomorphometry, scanning electron microscopy and energy dispersive X-ray analysis, and used for mathematical description of bone formation. The results showed that both the composition of the glass and the granule size of the granules, have influence on bone growth from the surrounding tissue. Glass S53P4, which from previous observations is known to be an effective bioactive glass and widely used in the Biomaterial Project of Turku, Finland, showed bone bonding and increasing bone growth between the granules. Glass S59.7P2.5 which due to its high Si-content should be inert, showed bone bonding. At 4 weeks the bone growth was significantly more abundant in bone defects filled with large granules (630 800 microm) than in defects filled with small granules (200-250 microm). Glass S52P3 with an alumina content of 3 wt %, showed good bone conduction, possibly even bone bonding for granules of 630-800 microm size. Granules of 200-250 microm with a high alumina content at the surface of the reaction layer, showed hardly any bone contact at all. This data, therefore, gives new information concerning bone bonding and osteoconduction of bioactive glasses with a high silica or alumina content. PMID- 15348462 TI - Investigation of the mechanical performance of young glass-ionomer cement using dynamic mechanical analysis. AB - Glass-ionomer (or more correctly, glass polyalkenoate) cements have wide applications in dentistry. This paper reports an investigation using dynamic mechanical analysis (DMA) on the setting of typical conventional glass-ionomer cements of varying age. Rectangular section cement samples were stored for four weeks in distilled water at 37 degrees C before being tested. The experimental procedure involved the clamping of the sample in tensile mode and heating through a 37-95 degrees C temperature range in water. A general behavioral trend was followed where all the samples showed increased flexibility with rise in temperature until a "threshold" temperature was reached, whereupon sudden tensile stiffening was observed. The temperature at which the stiffening took place was dependent on the age of the cement, and was interpreted in terms of the secondary cement forming reactions of silica and phosphate. The younger samples stiffened at significantly lower temperatures than the older ones. The activated nature of glass-ionomer setting chemistry meant that younger cements could be prematurely aged through heating. Differential scanning calorimetry was used to study the effect of heating on the distribution of loosely bound water in the cements that had, and had not been exposed to a DMA cycle. Most notably, it was seen that the DMA process did not affect the water in the matrix. This leant further credence to the hypothesis that the stiffening observed during the DMA heating process was caused by accelerated network formation. PMID- 15348463 TI - Minimum solid area models applied to the prediction of Young's modulus for cancellous bone. AB - In developing models for the mechanical behavior of cancellous bone, accurate prediction of Young's modulus as a function of the pore fraction and morphology is a requirement. Previous workers have suggested models which provide good statistical fits, but most of these models are highly idealized, with no treatment of the actual morphology of the porosity. In the field of engineering ceramics, simple minimum solid area models have been developed over the past four decades to describe the mechanical properties of porous structural ceramics. This paper applies these models to data for cancellous bone, and it is shown that one, developed specifically for high porosity materials, gives realistic predictions of tissue modulus and a good statistical fit to well-established data. This model should prove to be useful in biomechanical analyses involving cancellous bone tissue. PMID- 15348464 TI - Current state of the art of biphasic calcium phosphate bioceramics. AB - We have developed 15 years ago, with the collaboration of Lynch, Nery, and LeGeros in the USA, a bioactive concept based on biphasic calcium phosphate (BCP) ceramics. The concept is determined by an optimum balance of the more stable phase of HA and more soluble TCP. The material is soluble and gradually dissolves in the body, seeding new bone formation as it releases calcium and phosphate ions into the biological medium. The bioactive concept based on the dissolution/transformation processes of HA and TCP has been applied to both Bulk, Coating and Injectable Biomaterials. The events at the calcium phosphate (CaP) biomaterial/bone interface represent a dynamic process, including physico chemical processes, crystal/proteins interactions, cells and tissue colonization, bone remodeling, finally contributing to the unique strength of such interfaces. An important literature and numerous techniques have been used for the evaluation of the fundamental physico chemical and biological performance of BCP concept. This type of artificial bone used from a long time in preclinical and in clinical trial, revealed the efficiency for bone filling, performance for bone reconstruction and efficacy for bone ingrowth at the expense of the micro macroporous BCP bioceramics. PMID- 15348465 TI - Biphasic calcium phosphate bioceramics: preparation, properties and applications. AB - Biphasic calcium phosphate (BCP) bioceramics belong to a group of bone substitute biomaterials that consist of an intimate mixture of hydroxyapatite (HA), Ca(10)(PO(4))(6)(OH)(2), and beta-tricalcium phosphate (beta-TCP), Ca(3)(PO(4))(2), of varying HA/beta-TCP ratios. BCP is obtained when a synthetic or biologic calcium-deficient apatite is sintered at temperatures at and above 700 degrees C. Calcium deficiency depends on the method of preparation (precipitation, hydrolysis or mechanical mixture) including reaction pH and temperature. The HA/beta-TCP ratio is determined by the calcium deficiency of the unsintered apatite (the higher the deficiency, the lower the ratio) and the sintering temperature. Properties of BCP bioceramics relating to their medical applications include: macroporosity, microporosity, compressive strength, bioreactivity (associated with formation of carbonate hydroxyapatite on ceramic surfaces in vitro and in vivo), dissolution, and osteoconductivity. Due to the preferential dissolution of the beta-TCP component, the bioreactivity is inversely proportional to the HA/beta-TCP ratio. Hence, the bioreactivity of BCP bioceramics can be controlled by manipulating the composition (HA/beta-TCP ratio) and/or the crystallinity of the BCP. Currently, BCP bioceramics is recommended for use as an alternative or additive to autogeneous bone for orthopedic and dental applications. It is available in the form of particulates, blocks, customized designs for specific applications and as an injectible biomaterial in a polymer carrier. BCP ceramic can be used also as grit-blasting abrasive for grit-blasting to modify implant substrate surfaces. Exploratory studies demonstrate the potential uses of BCP ceramic as scaffold for tissue engineering, drug delivery system and carrier of growth factors. PMID- 15348466 TI - Mesenchymal stem cells combined with biphasic calcium phosphate ceramics promote bone regeneration. AB - The reconstruction and repair of large bone defects, resulting from trauma, cancer or metabolic disorders, is a major clinical challenge in orthopaedics. Clinically available biological and synthetic grafts have clear limitations that necessitate the development of new graft materials and/or strategies. Human mesenchymal stem cells (MSCs), obtained from the adult bone marrow, are multipotent cells capable of differentiating into various mesenchymal tissues. Of particular interest is the ability of these cells to differentiate into osteoblasts, or bone-forming cells. At Osiris, we have extensively characterized MSCs and have demonstrated MSCs can induce bone repair when implanted in vivo in combination with a biphasic calcium phosphate, specifically hydroxyapatite/tricalcium phosphate. This article reviews previous and current studies utilizing mesenchymal stem cells and biphasic calcium phosphates in bone repair. PMID- 15348467 TI - Bilayered calcium phosphate coating to promote osseointegration of a femoral stem prosthesis. AB - A bilayered bioactive-gradient coating, consisting of a superficial layer of biphasic calcium phosphate (BCP) and a deep layer of hydroxyapatite (HA), promotes faster osseointegration and higher shear strength in non-loading conditions than do monolayer BCP or HA coatings. This study evaluated the biofunctionality of this coating in weight-bearing conditions at 6 and 12 months. The coating was plasma-sprayed on the metaphyseal portion of a sandblasted Ti6Al4V canine femoral prosthesis implanted using the surgical press-fit technique. An identical uncoated stem served as the control. Metaphyseal bone-to implant apposition was increased for coated ( approximately 90% and 80% respectively at 6 and 12 months) as compared to uncoated implant ( approximately 7% at 6 and 12 months). Limited bone apposition was observed at the diaphyseal level. After 12 months, the uncoated implant interface consisted of well organized, active fibrous tissue, whereas only inactive fibrous tissue interposition was observed at diaphyseal levels of the coated implant. At 6 months, the mineralization apposition rate (MAR) was similar, regardless of implant or bone structures. At 12 months, a significant decrease of MAR was observed around the uncoated implant. Transmission electron microscopy studies of the interface showed precipitation of biological apatite crystals in close association with mineralized collagenous bone matrix. Our results suggest a direct relationship between bioactivity and enhanced bone formation. The sandwich coating used is effective in promoting massive metaphyseal osseointegration, which ensures mechanical stability for early weight-bearing and should prevent long-term complications. PMID- 15348468 TI - Micro macroporous biphasic ceramics and fibrin sealant as a moldable material for bone reconstruction in chronic otitis media surgery. A 15 years experience. AB - Bone reconstruction is still a matter of concern in middle ear surgery despite the large number of surgical techniques proposed. Combination of biphasic calcium phosphate bioceramics with human fibrin sealant forms a moldable material easy to apply for bone reconstruction. Since 1986, we have been using this composite for reconstruction of mastoid cavities during chronic otitis media surgery. Granules of ceramic provide immediate mechanical stability and later on, promote osseoinduction when fibrin sealant forms a strong bond between granules enhancing the wound healing process. After checking feasibility, efficacy and tolerance through a controlled study on dogs by filling mastoid cavities, we started a clinical series comprising yet 66 mastoid reconstructions. A retrospective data analysis on 63 patients with an average follow up of 42 months including 12 histological controls confirms the stability of bone reconstruction with a remarkable tolerance of the skin in contact. Biopsies confirmed progressive replacement of material by lamellar newly formed bone. Mastoid cavities have a randomed shape making complex bone reconstruction procedure and their filling by the composite constitutes an easy way and represents a highly satisfactory procedure. The authors considered that this should be explored in other indications of bone surgery. PMID- 15348469 TI - Tissue engineering of dermal substitutes based on porous PEGT/PBT copolymer scaffolds: comparison of culture conditions. AB - Previously, it was found that chondrocytes and fibroblasts could be efficiently seeded onto three-dimensional scaffolds in spinner flasks. In this study different culture conditions were compared to create a living dermal substitute as rapidly as possible. Human dermal fibroblasts were dynamically seeded onto biodegradable porous PEGT/PBT copolymer (PolyActive) scaffolds for 24 h in spinner flasks. Subsequently, the cell-seeded scaffolds were cultured in two conditions: statically (without medium flow, S) and dynamically (with slow medium flow, D). Qualitative analyses (scanning electron microscopy and histology) and quantitative assays for DNA, total collagen (hydroxyproline) and glycosaminoglycans were done with samples cultured for 3, 7, 14 and 21 days. In dynamically cultured constructs, human dermal fibroblasts were uniformly distributed throughout the pores of the scaffolds and had deposited higher amounts of extracellular matrix (ECM). Significantly higher numbers of fibroblasts were found (p<0.001), and significantly more collagen (hydroxyproline content) (p<0.001) and glycosaminoglycan (GAG) (p<0.05) were deposited at all the investigated time points when compared to static cultured constructs. In conclusion, medium flow stimulated the proliferation of human dermal fibroblasts and accelerated the ECM deposition in PolyActive dermal substitutes when compared to static culture. Dynamic culture reduced the time to create a dermal substitute containing autologous fibroblasts. PMID- 15348470 TI - Surface oxidation of UHMWPE for orthopedic use increases apoptosis and necrosis in human granulocytes. AB - Ultra high molecular weight polyethylene (UHMWPE) used in orthopedic prosthesis is often sterilized with gamma-rays and the subsequent oxidation was suggested to favor the in vivo wear. UHMWPE debries produced by wearing trigger an inflammatory response that can led to the implant failure. To explore direct effects of UHMWPE oxidation on immunocompetent cells and their possible role in the prosthesis failure, peripheral blood cells (PBCs) have been grown for 24 and 48 h onto plastic (Ct), UHMWPE (PE) and heat oxidized UHMWPE (PEOx) discs. PBCs necrosis and apoptosis were assessed in flow cytometry using propidium iodide staining. After 24 h, no statistically significant differences were observed in the amount of apoptotic and necrotic cells between Ct, PE and PEOx samples while, after 48 h, both necrotic and apoptotic cells were strongly increased in PEOx samples where also the granulocytes population appeared strongly reduced (6.3+/ 1.1%) compared to PE (10.5+/-1.5%) and Ct (15.1+/-0.9%) samples. We conclude that surface oxidation may interfere with prosthetic failure and/or integration via granulocytes modulation. PMID- 15348471 TI - Titanium implants and BMP-7 in bone: an experimental model in the rabbit. AB - This study evaluates the effect of rhBMP-7/OP-1 on the osseointegration of commercially pure titanium implants in an experimental implant model in rabbits. Threaded titanium implants with two transverse parallel canals were inserted in the femur and tibia of rabbits. The canals were filled with, 10 microg of BMP 7/collagen carrier, pure collagen carrier or were left empty as a control. The stiffness of the implant fixation was evaluated by Resonance Frequency Analysis (RFA) at baseline and four weeks postoperatively. Percentage of bone ingrowth in the canals was measured on microradiographs. Histomorphometry along the threaded part of the implants was performed on 15 microm thin sections. The results from the RFA demonstrated a higher mean value for the BMP-7 treated implants in the tibia than the carrier treated implants but not compared to the control implants. The control implants in the tibia demonstrated more bone ingrowth in the upper canal than to the carrier or the BMP-7 treated implants. Apart from these differences there were no significant effects of BMP. In this study BMP-7 did not contribute to any substantially improved bone anchorage of titanium implants. PMID- 15348472 TI - Gelatin manipulation of latent macropores formation in brushite cement. AB - Macroporous brushite cement was prepared from a mixture of beta-tricalcium phosphate (beta-TCP) and monocalcium phosphate monohydrate (MCPM) using gelatin powder as a latent templates. In a setting reaction coexisting with gelatin, closed packed, open-pore structure with 100-200 microm macropores are obtained after immersion of the set cement into PBS buffer (pH 7.4) at 37 degrees C for 1 4 weeks. The macroporous brushite cement has compressive strength of 15 MPa originally, which reducing to 5.5 MPa with macropore formation gradually in comparison to that of cancellous bone (5-10 MPa). PMID- 15348473 TI - Modification of human polymorphonuclear neutrophilic cell (PMN)-adhesion on biomaterial surfaces by protein preadsorption under static and flow conditions. AB - Biomaterials induce a specific reaction after implantation in the human body. This reaction depends on the chemical and physico-chemical properties of the material as well as on the site and type of implantation. We have used a dynamic model, the parallel-plate flow-chamber, to examine the interactions of different biomaterials with polymorphonuclear neutrophilic cell (PMN) and how these interactions are influenced by protein preadsorption. Our results clearly show that for hydrophobic materials, glass and PE, which induce a prominent adhesion of PMN, the mixture of albumin and fibrinogen induces the best inhibitory effect. On hydrophilic biomaterial surfaces, untreated TCPS and PC-coated TCPS, reveal only a minor influence of adsorbed proteins on PMN adhesion because of a primary low adhesive surface for PMN and proteins as well. Human citrated plasma leads only to a slight inhibition of PMN adhesion. On the hydrophobic materials, glass and PE, bovine serum albumin (BSA) had the best anti-adhesive potential with respect to PMN. The coating using phosphorylcholine is an excellent surface modification to prevent PMN-adhesion and protein adsorption. The results of our experiments suggest that investigations under static and flow conditions are also needed to determine the influence of protein adsorption on other relevant blood cell populations, for example, platelets and monocytes. PMID- 15348474 TI - The response of macrophages to particles of resorbable polymers and their degradation products. AB - Alpha polyesters such as poly(L-lactide) and poly(glycolide) are biodegradable materials used in fracture fixation and they need to be assessed for problems associated with their degradation products. This study has compared cell responses to low molecular weight poly(L-lactide) particles, lactate monomer, poly(glycolide) particles and glycolic acid at cytotoxic and sub-cytotoxic concentrations. Murine macrophages were cultured in vitro and the release of lactate dehydrogenase (LDH), prostaglandin E(2) (PGE(2) and interleukin-1 alpha IL-1alpha was measured following the addition of particles or monomer. Experiments revealed that both the poly(L-lactide) and poly(glycolide) particles gave rise to dose dependent increases in LDH release and an increase in IL-1alpha and PGE(2) release. Comparisons of the poly(L-lactide) particles to the poly(glycolide) particles did not reveal any differences in their stimulation of LDH, IL-1alpha and PGE(2) release. The lactate and glycolate monomers did not increase PGE(2) or IL-1alpha release above control levels. There was no difference in biocompatibility between the poly(L-lactide) and poly(glycolide) degradation products both in particulate and monomeric form. PMID- 15348475 TI - The effect of varying percentage hydroxyapatite in poly(ethylmethacrylate) bone cement on human osteoblast-like cells. AB - Poly(ethylmethacrylate) (PEMA) bone cement has been developed, and the cements mechanical properties are improved by the incorporation of particulate fillers, such as hydroxyapatite (HA). In this in vitro study, human osteoblast-like (HOB) cells were used to examine the effect on cellular behavior of the addition of HA to PEMA using a plain PEMA control. Thymidine uptake ((3)H-TdR) and total DNA were used to assess cell growth and proliferation. Confocal laser scanning microscopy (CLSM) was used to study focal contacts and actin cytoskeletal organisation. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to assess cell morphology and cellular ultrastucture. The early time points showed preferential anchorage to the HA exposed on the cement surface, but no difference in adhesion or proliferation. These results have been attributed to increases in residual monomer with HA incorporation, as shown by proton nuclear magnetic resonance (H(1)-NMR) spectra. PMID- 15348476 TI - Preparation of Gd3+-containing polymer complex as a novel magnetic resonance signal-enhancing coating material. AB - This study investigated the suitability of a Gd(3+)-containing polymer complex used as a magnetic resonance (MR) signal-enhancing coating material. The synthesis of the Gd>(3+)-containing polymer complex was done by conjugating N-(2 hydroxyethyl)ethylenediaminetriacetic acid (HEDTA) with poly (styrene-maleic acid) copolymer (SMA), followed by coordinating with Gd(3+). The characterizations of FT-IR, NMR and XPS confirmed that HEDTA was covalently attached to SMA and Gd(3+)-containing polymer complex was formed. Coating such Gd(3+)-containing polymer complex on the surface of polypropylene (PP) catheter was performed by solution coating method. The results of MR imaging experiments in vitro indicated that the PP catheter coated with this coating material showed strong MR signal in the vicinity of the catheter surface and a clear contrast between the catheter surface and background. Based on these results, a novel MR signal-enhancing coating material, which showed the great potentialities in making catheters for endovascular intervention, visible by MR imaging, can be developed. PMID- 15348479 TI - Nerve growth factor beta(NGF beta) delivery via a collagen/hydroxyapatite (Col/HAp) composite and its effects on new bone ingrowth. AB - In craniofacial surgery, bone is needed to augment misshapen areas and to fill gaps during repair of congenital anomalies and injuries resulting into bone deficiencies. Examples of conditions requiring bone tissue include missing alveolar bone in cleft palates, bony nasal pyramid defects following removal of fistulous tracts or cysts and defects following removal of sinus and mandibular tumors. Moreover, maxillofacial neurosensory deficiencies may be caused by various surgical procedures, such as tooth extraction, osteotomies, pre prosthetic procedures, excision of tumors or cysts, surgery of TMJ, and surgical treatment of fractures and cleft lip/palate. Therefore, a tissue engineering approach to craniofacial surgery has a crucial importance: the use of various composites with osteoconductive ceramics, polymers, bioactive factors, cells, or a combination of them, offers the possibility of rapid tissue regeneration and integration with the host tissue. In this study, a composite consisting of two well-known biomaterials, collagen/hydroxyapatite (Col/HAp), was used as a drug delivery device for neurotrophin - nerve growth factor beta (NGF beta). This delivery device, enriched with neurogenic-osteogenic factor, was analyzed in vitro and in vivo. It was implanted into calvaria defects of 20 Wistar rats, weighing 200-250 g. Implants were left in place for different periods of time. Controls were as follows: (a) contralateral defect without any implant; and (b) contralateral defect implanted with composite without NGF factor. The rats were euthanized after 30 days, and the implant sites and explants were examined clinically, histologically, SEM and histomorphometrically. Our results evidenced stimulation of periosteal and endocortical woven and lamellar bone formation, with increases in bone mass and decreases in bone marrow. We found that NGF enhanced the remodeling activity in the intracortical region, and induced an increase in the intracortical cavity number and area by the end of the study. In vitro results were in line with in vivo ones. We believe that the composite proposed in this study has considerable advantages in tissue engineering and is very suitable as a biomaterial for the filling of irregular defects in maxillo facial surgery. Two areas of clinical research will be impacted by this system. The first is pharmaceutical research on drug delivery and high-throughput screening of neurotrophic-osteogenic compounds. Transplantation research is the second area that will benefit from the system. PMID- 15348480 TI - Development and properties of polycaprolactone/hydroxyapatite composite biomaterials. AB - Polycaprolactone/hydroxyapatite (PCL/HA) composites were prepared by two different procedures. The first one consists of a conventional blending of the polymer and the reinforcement material in an extruder. The second method consists of grafting of PCL on the surface of HA particles. This was achieved by a ring opening polymerization of caprolactone in the presence of HA, where its OH groups act as initiators. By this method, it was possible to obtain, in one step, a composite of PCL and surface modified HA. In both methods different percentages of filler were used to obtain several composites. These composites were characterized with respect to their mechanical properties, in the dry and wet state, by means of tensile tests on compression molded samples. The polymer/filler interface was analyzed by scanning electron microscopy. Water uptake and weight loss degradation experiments were also performed. An increase in the modulus for higher amounts of filler was, as expected, observed in the composites obtained by both processes. Furthermore, the mechanical properties of the materials in the wet state are considerably lower than those in the dry state. However, this difference is more significant for the composites obtained by conventional blending than for composites obtained by the grafting procedure, indicating that the later procedure can be an adequate route to reduce water susceptibility of PCL/HA composites. PMID- 15348481 TI - Development of a polymer stent with shape memory effect as a drug delivery system. AB - The article presents a new concept for vascular endoprothesis (stent). Almost all commercially available stents are made of metallic materials. A common after effect of stent implantation is restenosis. Several studies on metal stents coated with drug show, that the use of a drug delivery system may reduce restenosis. The purpose of this work is to develop a new stent for the drug delivery application. The shape memory properties of thermoplastic polyurethane allow to design a new fully polymeric self-expandable stent. The possibility to use the stent as a drug delivery system is described. PMID- 15348482 TI - Preliminary study of a polycaprolactone membrane utilized as epidermal substrate. AB - Solvent-cast sheets of polycaprolactone were biaxially stretched to produce 10-15 microm thick films. PCL films were found to have a tensile strength of 55 MPa which is about two and a half times stronger than native skin. One of our previous studies using non-coated PCL membranes showed that only 36% of the membrane surface was covered with keratinocytes after 9 days of culture. The present study examined the effects of coating the surface of PCL membranes with fibrin on the proliferation of keratinocytes. Qualitative analysis revealed that the cells attached and proliferated better on coated PCL films. Keratinocytes exhibited healthy cobblestone morphology and proliferated as continuous monolayers over a period of 16 days. The results indicated that fibrin coated PCL films would support the attachment and proliferation of human keratinocytes and have the potential to be applied as a matrix material for tissue engineering an epidermal equivalent. PMID- 15348483 TI - Poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) based tissue engineering matrices. AB - In this study, the aim was to produce tissue-engineered bone using osteoblasts and a novel matrix material, poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV). In order to prepare a porous PHBV matrix with uniform pore size, sucrose crystals were loaded in the foam and then leached leaving pores behind. The surface of the PHBV matrix was treated with rf-oxygen plasma to increase the surface hydrophilicity. SEM examination of the PHBV matrices was carried out. Stability of PHBV foams in aqueous media was studied. The pH decrease is an indication of the degradation extent. The weight and density were unchanged for a period of 120 days but then a significant decrease was observed for the rest of the study. Osteoblast cells were then isolated from rat bone marrow and seeded onto PHBV matrices. The metabolization and proliferation on the foams was determined with MTS assay which showed that osteoblasts proliferated on PHBV. It was also found that cells proliferated better on large pore size foams (300-500 microm) than on the small pore size foams (75-300 microm). Production of ALP was measured spectrophotometrically. The present study demonstrated that PHBV matrices are suitable substrates for osteoblast proliferation and differentiation. PMID- 15348484 TI - Thermal properties of thermoplastic starch/synthetic polymer blends with potential biomedical applicability. AB - Previous studies shown that thermoplastic blends of corn starch with some biodegradable synthetic polymers (poly(epsilon-caprolactone), cellulose acetate, poly(lactic acid) and ethylene-vinyl alcohol copolymer) have good potential to be used in a series of biomedical applications. In this work the thermal behavior of these structurally complex materials is investigated by differential scanning calorimetry (DSC) and by thermogravimetric analysis (TGA). In addition, Fourier transform infrared (FTIR) spectroscopy was used to investigate the chemical interactions between the different components. The endothermic gelatinization process (or water evaporation) observed by DSC in starch is also observed in the blends. Special attention was paid to the structural relaxation that can occur in the blends with poly(lactic acid) at body temperature that may change the physical properties of the material during its application as a biomaterial. At least three degradation mechanisms were identified in the blends by means of using TGA, being assigned to the mass loss due to the plasticizer leaching, and to the degradation of the starch and the synthetic polymer fractions. The non isothermal kinetics of the decomposition processes was analyzed using two different integral methods. The analysis included the calculation of the activation energy of the correspondent reactions. PMID- 15348485 TI - Novel calcium phosphate composite bone cement: strength and bonding properties. AB - A new high-strength cement prepared from calcium phosphate and calcium aluminate has been developed and was evaluated for potential use in bone and joint repair applications. Cement specimens were aged under simulated physiological conditions. The compressive strength of the cement was determined at time intervals 1 h after setting up to 52 weeks. A compressive strength of 111.6+/ 12.9 MPa was measured at 4 weeks, with the cement attaining 64% of this maximum strength within 4 h of preparation. Compressive strength greater than 90 MPa was maintained up to 52 weeks. The strength of the cement-prosthesis interface was studied using a pull-out test. Polished, 316L stainless steel rods were implanted in canine cadaver femurs to simulate a cemented hip prosthesis. At 4, 24 h, and 60 days post implantation, the force required to displace the rod was measured. Mean interfacial shear strengths of 1.17+/-0.25, 1.11+/-0.21, and 1.11+/-0.32 MPa were observed at respective time-periods. PMID- 15348486 TI - Functionalized surfaces of polylactide modified by Langmuir-Blodgett films of amphiphilic block copolymers. AB - To modify the surface of poly(L-lactide) (PLA) supports, we have investigated the feasibility to deposit on the PLA surface Langmuir-Blodgett films of amphiphilic block copolymers based on poly(L-lactide). AB and ABA block copolymers were prepared with PLA as the A block and either poly(ethylene oxide), alpha-methoxy omega-hydroxy poly(ethylene oxide), alpha-carboxy-omega-hydroxy poly(ethylene oxide) or poly(L-aspartic acid) as the B blocks. Films with phase-separated hydrophilic and hydrophobic blocks in a bilayer "brush" structure were prepared by compression of the copolymer Langmuir films on the water/air interface. The interfacial behavior of the monolayers and the effect of the copolymer composition on the phase separation was followed by measurements of the surface pressure/area isotherms using a Langmuir trough and by contact angle measurement of deposited Langmuir-Blodgett (LB) films. The phase separation of the hydrophilic and PLA blocks is more effective in diblock AB copolymers compared with triblock ABA copolymers. The presence of ionic groups in the hydrophilic chains facilitates penetration of hydrophilic segments into the water subphase. Dynamic contact angle measurements were used to study the stability of the LB films transferred on the PLA support and the changes in the surface properties upon incubation of surfaces in water. PMID- 15348487 TI - Properties of calcium phosphate coatings deposited and modified with lasers. AB - Physical, chemical and biological properties of calcium phosphate coatings fabricated by a pulse laser deposition method at room temperature (RT PLD) have been studied. In vitro evaluation of RT PLD coatings on bioresorbable polymers (Poly-epsilon-caprolactone and Poly-L-lactide) have been carried out. It was shown that both polymers support osteoblast growth, with increased cell activity, alkaline phosphatase activity and total protein content on those surfaces that have been coated. The advantages of RT PLD coatings in biomaterials surface optimization are discussed. PMID- 15348488 TI - Preliminary study on human protein adsorption and leukocyte adhesion to starch based biomaterials. AB - In this study, the adsorption of human serum albumin (HSA), fibronectin (FN) and vitronectin (VN) onto the surface of novel biodegradable materials was evaluated by immunostaining. Specifically, polymeric blends of corn starch with cellulose acetate (SCA), ethylene vinyl alcohol copolymer (SEVA-C), and polycaprolactone (SPCL) were immersed in unitary and competitive systems; that is, binary and more complex protein solutions. For binary solutions, different HSA and FN protein distribution patterns were observed depending on the starch-based surface. Furthermore, the relative amount of proteins adsorbed onto starch-based surfaces was clearly affected by protein type: a preferential adsorption of VN and FN as compared to HSA was observed. On tests carried out with unitary, binary and more complex solutions, it was found that vitronectin adsorption ability was enhanced in competitive systems, which was associated with a lower amount of adsorbed albumin. In order to assess the effect of these human proteins on cell behavior, a mixed population of human lymphocytes and monocytes/macrophages was cultured over pre-coated SEVA-C surfaces. Through anti-CD3 and CD-14 monoclonal antibody labeling and cell counting, leukocyte adhesion onto pre-coated SEVA-C surfaces was analyzed. Based on the results, it was possible to detect albumin long-term effects and fibronectin short-term effects on cell adhesion proving that previously adsorbed proteins modulate leukocyte behavior. PMID- 15348489 TI - Evaluation of the potential of starch-based biodegradable polymers in the activation of human inflammatory cells. AB - The inflammatory response resulting from the implantation of a medical device may compromise its performance and efficiency leading, in certain cases, to the failure of the implant. Thus, the assessment of the behavior of inflammatory cells in vitro, constitutes a key feature in the evaluation of the adverse potential, or not, of new promising biomaterials. The objectives of this study were to determine whether starch-based polymers and composites activated human neutrophils. Blends of starch with ethylene-vinyl alcohol, with cellulose acetate and polycaprolactone, as well as composites based on all these materials filled with hydroxyapatite have been studied. A lysozyme assay was adapted to examine enzyme secretion from human neutrophils incubated with different starch-based materials. Changes in the free radical and degranulation activity of the neutrophil were also determined by measuring the luminescent response of Pholasin, a photoprotein that emits light after excitation by reactive oxygen species. The amount of lysozyme secreted by neutrophils incubated with the polymers did not exhibit significant differences between the tested materials. Results were in all cases similar to those obtained for the control (polypropylene) except for one of the starch blends (corn starch with polycaprolactone reinforced with 30% (w/w) of HA). The chemiluminescence experiments showed that polymers reduce the signal produced by activated neutrophils. Furthermore, for some polymers it was demonstrated that the phenomenon was due to an effect of the surface of the materials in cell adhesion or a simultaneous competition for the photoprotein in solution, which results in the decrease of the intensity of light emitted and detected. PMID- 15348490 TI - Novel reverse thermoresponsive injectable poly(ether carbonate)s. AB - The water solutions of polymers displaying reverse thermal gelation (RTG), such as poly(ethylene oxide)/poly(propylene oxide)/poly(ethylene oxide) triblocks, exhibit a pronounced viscosity increase as temperature rises, within a very narrow temperature interval. Unfortunately, the viscosity increase attained by these solutions is not large enough, resulting in systems displaying limited stability and short residence times. This paper introduces a new family of reverse thermoresponsive alternating [A-B](n) block copolymers, comprising poly(ethylene oxide) (PEO), and poly(propylene oxide) (PPO) chains, using phosgene as the molecule connecting both components. The effect of various compositional and structural parameters on both the Ci (minimal gelation concentration) and Ti (minimal gelation temperature) of these systems was investigated. The copolymers were characterized by GPC, (1)H-NMR, FT-IR, and DSC and the rheological behavior of the water solutions was studied using a Brookfield viscometer. The water systems were also studied by dynamic light scattering (DLS) and fluorescence spectroscopy. The copolymers developed exhibited clearly superior rheological properties, when compared to existing RTG displaying PEO-PPO-PEO triblocks. For example, while the viscosity of a 15% water solution of the commercially available Pluronic F-127 achieved 5000 Pa.s, at 37 degrees C, poly(ether carbonate) water solutions (15%) attained viscosities between 25 000 and 150 000 Pa.s. PMID- 15348491 TI - Introducing lactide-based biodegradable tissue adhesives. AB - Lactide-based low molecular weight copolymers were synthesized and investigated as tissue adhesives. The oligomers were composed of di or trifunctional central connecting segments and lateral PLA blocks. Copolymers with glass transition temperatures in the 20-25 degrees C range, were found to perform better. Strong connection was found between the length of the PLA blocks, the glass transition temperature (T(g)) and the Adhesive Failure Strength of the different materials. Flexible epsilon-caprolactone (CL) molecules were inserted into the PLA blocks, to produce longer biodegradable chains and improve the adhesive strength of the oligomers, while keeping their T(g) within the appropriate temperature interval. Branched oligomers consisting of a trimethylolpropane central molecule and three LA-CL segments, displayed enhanced in vitro adhesive properties. PMID- 15348492 TI - Plasma- and chemical-induced graft polymerization on the surface of starch-based biomaterials aimed at improving cell adhesion and proliferation. AB - Plasma and chemical induced graft polymerization of acrylic monomers on starch based biomaterials has been performed with the aim to improve cell adhesion and proliferation on the surface of the polymers, in order to adequate their properties for bone tissue engineering scaffolding applications. Plasma and chemical surface activation was aimed to induce the polymerization of acrylic polar monomers being carried out by applying a radio frequency plasma and expose the samples to a mixture of Ar/O2, or by immersion in a H2O2/(NH4)2S2O8 solution with UV radiation, respectively. Both procedures were followed by the graft polymerization of the corresponding monomers. Polymer grafting was analyzed by Fourier transformed infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) and by contact angle measurements. Properties such as mechanical performance, swelling degree, and degradation behavior, as well as bioactivity, have been studied and compared for the different activation methods. Finally, preliminary cell adhesion and proliferation tests were performed, using goat bone marrow cells, showing a remarkable improvement with respect to original non-surface modified starch-based biomaterials. PMID- 15348493 TI - Factors affecting the optical properties of Pd-free Au-Pt-based dental alloys. AB - The optical properties of experimental Au-Pt-based alloys containing a small amount of In, Sn, and Zn were investigated by spectrophotometric colorimetry to extract factors affecting color of Au-Pt-based high-karat dental alloys. It was found that the optical properties of Au-Pt-based alloys are strongly affected by the number of valence electrons per atom in an alloy, namely, the electron:atom ratio, e/a. That is, by increasing the e/a-value, activities of reflection in the long-wavelength range and absorption in the short-wavelength range in the visible spectrum apparently increased. As a result, the maximum slope of the spectral reflectance curve at the absorption edge, which is located near 515 nm (approximately 2.4 eV), apparently increased with e/a-value. Due to this effect, the b*-coordinate (yellow-blue) in the CIELAB color space considerably increased and the a*-coordinate (red-green) slightly increased with e/a-value. The addition of a third element with a higher number of valence electrons to the binary Au-Pt alloy is, therefore, effective in giving a gold tinge to the parent Au-Pt alloy. This information may be useful in controlling the color of Au-Pt-based dental alloys. PMID- 15348494 TI - Bioactivity of metallic biomaterials with anatase layers deposited in acidic titanium tetrafluoride solution. AB - A simple and versatile treatment was developed to provide various metallic biomaterials such as Ti, NiTi, Ta and SUS 316L stainless steel with in vitro bioactivity or ability to deposit carbonate-incorporated apatite in a simulated body fluid (Kokubo solution). A well-crystallized anatase layer deposited on the metallic biomaterials surfaces after soaking them at 60 degrees C for 24 h in an aqueous solution of titanium tetrafluoride (40 mM) whose pH was adjusted to 1.9 with HCl. The as-coated anatase layers did not deposit apatite. When heated at 300 degrees C they were so bioactive as to deposit apatite within 5 day(s) in the Kokubo solution. The trace amount of fluorine weakly bound in the as-coated anatase layers was suggested to be one of the factors that suppressed the bioactivity. PMID- 15348495 TI - Mechanical properties of a permanent dental restorative material based on calcium aluminate. AB - This paper deals with some important mechanical properties (hardness, dimensional stability, compressive and flexural strength) of an experimental version of a translucent calcium aluminate dental restorative material. All samples investigated have been made from pre-pressed tablets, with a compaction degree of approximately 60%, hydrated using a 0.15 wt % Li salt solution as an accelerator. The samples were stored in water at 37 degrees C between the measurements. As reference materials one composite, Tetric Ceram, and one glass ionomer, Fuji II, were used with specimens prepared according to the manufacturer's recommendations. For the reference materials some of the properties were published data. The results show that the calcium aluminate material has sufficient mechanical properties to be used as a permanent dental restorative taking as a reference the ISO 9917 and the ISO 4049 as well as the reference materials. In addition the results indicate that the mechanical properties are controlled by the microstructure, which is mainly determined by the grain size of the filler. PMID- 15348496 TI - Controllable porosity hydroxyapatite ceramics as spine cage: fabrication and properties evaluation. AB - A procedure was designed to prepare porosity-graded hydroxyapatite (HA) ceramics simulating the bimodal structure of natural bone, which could be used to build a cage that would promote the reconstruction of the anterior column after vertebrectomy or corpectomy in tumor and trauma surgery. HA ceramics with controllable pore size distribution and porosity were developed by using chitosan and Poly(vinyl alcohol) (PVA) as the pore-forming agents. HA ceramics with worthwhile properties such as a wide range of volume porosity (10-50%) and pore size (nanometer to 400 microm) can be obtained from this method, which allows the fabrication of HA ceramics with desirable porous characteristics simulating the bimodal natural bone architecture expected to provide advantages for bony fusion in the intervertebral foramina. When coated with chitosan-gelatin network, the bending strength of the porous HA ceramics significantly improved. The polymer network coated porous HA have potential application in the construction of cages for spinal operations. PMID- 15348497 TI - Time-dependent strength and fatigue resistance of dental direct restorative materials. AB - Elastic modulus (EM), initial fracture strength (FS) and flexural fatigue limit (FFL) of dental restorative materials were measured in a simulated oral environment to correlate mechanical response under the influence of water with the chemical nature of the test materials under investigation. One resin composite (RC; Tetric Ceram, Ivoclar-Vivadent Corp., Liechtenstein), an ion leaching resin composite (ILRC; Ariston pHc, Ivoclar-Vivadent Corp., Liechtenstein) a compomer (CO; Dyract AP, Dentsply Corp., USA) and a glass ionomer cement (GIC; Ketac Molar, 3MEspe Corp., Germany) were tested. Static EM, FS and dynamic FFL experiments were performed. The FFL was determined under cyclic loading for 10(5) cycles in terms of a staircase approach. The materials were stored for 1, 8, 30, 90 and 180 days in 37 degrees C distilled water, respectively. The RC degraded over time due to water adsorption followed by failure within the resin matrix. The ILRC suffered from a pronounced decrease in FS as well as in FFL due to a constant ion-leaching and macroscopic crack growth. CO failed over time due to resin-filler interface cracking. The GIC exhibited improved mechanical performance over time due to a post-hardening mechanism. The results reveal the necessity for substantial preclinical evaluation of direct restorative materials. The material parameters under investigation are capable of predicting clinical performance over time. PMID- 15348498 TI - In vitro degradation and cytocompatibility evaluation of novel soy and sodium caseinate-based membrane biomaterials. AB - Soy- and casein-based membranes are newly proposed materials disclosing a combination of properties that might allow for their use in a range of biomedical applications. Two of the most promising applications are drug delivery carrier systems and wound dressing membranes. As for all newly proposed biomaterials, a cytotoxic scanning must be performed as a preliminary step in the process of the determination of the compatibility with biological systems (biocompatibility). In this study, the cytotoxicity of both soy- and casein-based protein biomaterials has been evaluated and correlated with the materials degradation behavior. It was possible to show, through morphological and biochemical tests that these natural origin materials do not exert any cytotoxic effect over cells, and in some cases can in fact enhance cell proliferation. The different treatments to which the membranes were subjected during their processing (that include crosslinking with glyoxal and tannic acid, and physical modification by thermal treatment) seemed to have a clear effect both on the materials mechanical properties and on their in vitro biological behavior. PMID- 15348499 TI - Apatite formation on CaO-free polydimethylsiloxane (PDMS)-TiO2 hybrids. AB - Polydimethylsiloxane (PDMS)-TiO(2) hybrids with PDMS (M=550)/tetraethylorthotitanate molar ratios at 0.27, 0.68 and 1.35, i.e. Si/Ti atomic ratios at 2, 5 and 10 (hybrids PD2, PD5 and PD10, respectively) were prepared by a sol-gel method. Hybrid PD2 formed many cracks. Hybrids PD5 and PD10 were subjected to hot-water treatment 80 degrees C for 7 d. Hybrid PD5 produced cracks, whereas hybrid PD10 was crack-free after the hot-water treatment. Hybrid PD10 took a homogeneous amorphous structure before the hot-water treatment, and precipitated anatase particles 10-20 nm in size after the hot-water treatment. Hybrid PD10 did not form apatite on its surface in a simulated body fluid before the hot-water treatment, but formed it after the hot-water treatment. The obtained hybrid showed elastic deformation as large as 200% after the hot-water treatment. This kind of hybrid could be useful as a new type of bone-repairing material. PMID- 15348500 TI - Biocompatibility and biodegradation of poly(hydroxybutyrate)/poly(ethylene glycol) blend films. AB - Using chloroform as co-solvent, a series of poly(3-hydroxybutyrate) (PHB) and polyethylene glycol (PEG) blend materials with different ratio ranging from 80 : 20 (wt %) to 20 : 80 (wt %) were prepared by solution blend. The blood compatibility was evaluated by means of platelet clotting time test and exploring its morphological changes. The results showed that PEG played an important role in resisting platelet adhesion. With the increased addition of PEG, the clotting times became longer and the number of platelet adhesion decreased apparently. All platelets were in discrete state, no pseudopodium had been found and no collective phenomenon had been happened. The cell-compatibility was evaluated via Chinese Hamster Lung (CHL) fibroblast cultivation in vitro. The cells cultured on the matrix spread and proliferated well. With the increase of PEG content in the blend films, the number of live cells became more and more. These results indicated that PHB exhibited satisfying cell-compatibility and the addition of PEG also could improve the cell-compatibility of PHB. The biodegradation experiment indicated that the degradation of PHB/PEG was accelerated by enzyme in vitro and the blending of PEG was favorable to degradation. PMID- 15348501 TI - Preparation and characterization of pH sensitive sugar mediated (polyethylene glycol/chitosan) membrane. AB - Novel biodegradable membrane based on chitosan matrix was prepared and characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and swelling test. Native sugar, which is commonly used in human life, was utilized to prepare the crosslinked hydrophilic chitosan-polyethylene glycol (PEG) polyblend. According to TGA and FTIR results, the chemical reaction occurred in imine bonds (C=N) between sugar and amino groups in chitosan. Chitosan blending with high swelling capacity of PEG increased the water affinity and reacted with sugar decreased the water affinity. The equilibrium water content (EWC) value of the sugar mediated membrane is in the sequence of sucrose>D-fructose>glucose and they all present much lower water uptake ability than polyblend. The chitosan was not degraded by lysozyme, but all of the sugar-mediated membranes were susceptible to lysozyme. Scanning electron microscope (SEM) morphology shows that the degradation rate not only was controlled by the chemical complexation between sugar and polyblends, but the surface morphology of membranes also has great influence. Sucrose-mediated membrane supports the attachment and growth of NIH 3T3 fibroblasts. The pH sensitive and well degradable property of the sucrose-mediated membrane can be applied for biomedical application. PMID- 15348502 TI - Effect of micro- and macroporosity of bone substitutes on their mechanical properties and cellular response. AB - The control of porosity morphology and physico-chemical characteristics of calcium phosphate bone substitutes is a key-point to guaranty healing success. In this work, micro- and macroporosity of materials processed with 70% Hydroxyapatite (HAP) and 30% beta-tricalcium phosphate (beta-TCP) were controlled by sintering temperature and porogen addition, respectively. Porosity was quantified by scanning electron microscopy (pore size) and mercury intrusion porosimetry (interconnection between pores). The content of macrointerconnections and their size were dependent on porogen content, shape, and size. Mechanical properties (compressive strength) were strongly dependent on macroporosity size and content, on the basis of exponential laws, whereas microporosity ratio was less influent. Relying on those results, three types of materials with contrasting porous morphologies were processed and assessed in vitro, in primary culture of human osteoblasts and fibroblasts. With both types of cells, an exponential cellular growth was effective. Cells colonized the surface of the materials, bridging macroporosity, before colonizing the depth of the materials. Cell migration across and into macroporosity occurred via the emission by the cells of long cytoplasmic extensions that hanged on microporosity. Both macroporosity and macrointerconnectivity size influenced the penetration of cells. An interconnection size of 15 microm appeared to be effective to support this invasion without bringing down mechanical strength. PMID- 15348503 TI - Titanium transport through the blood stream. An experimental study on rats. AB - Different metals are increasingly being used to manufacture implants, especially in the fields of dentistry and orthopedics. No metal or alloy is completely inert in vivo. The metal and the organic fluids interact releasing, for example, metallic products. Several hypotheses regarding the probable dissemination routes of titanium have been postulated, but its valence, the organic nature of its ligands and its potential toxicity have yet to be established. In a previous experimental study we demonstrated that i.p. injected titanium and zirconium oxides disseminate and deposit in organs such as liver and lung. The aim of this work was to study the eventual participation of blood cells in the transport mechanism of titanium employing the intraperitoneal injection of titanium oxide in rats as the experimental model. Twenty male Wistar rats, x: 100 g body weight, were intraperitoneally injected with 16x10(3) mg/kg b.w. of TiO(2) in saline solution. Blood samples were taken by heart puncture at 3 and 6 months; blood smears were performed and stained with safranin evidencing monocytes containing titanium particles. The results obtained in this study would indicate that one of the ways in which titanium is disseminated is through the blood stream, via blood cells. PMID- 15348504 TI - Bone cements and fillers: a review. AB - Charnley [1] developed the first bone cement in the 1960s using poly(methyl methacrylate) (PMMA), which remains the most widely used material for fixation of orthopaedic joint replacements. In the field of dentistry, zinc polycarboxylate and glass polyalkenoate cements received major research interest from the 1970s to the present day. The discovery of a well-integrated intermediate layer between bone and many bioactive ceramic phases from the calcium-phosphate system, such as hydroxyapatite (HA), resulted in the development of new cements incorporating such phases. These investigations ranged from the development of castable bioactive materials to modified bioactive composites. This paper attempts to give a broad overview of the many different types of cements that have being developed in the past and those which are being researched at the present time. It has lead to a set of fundamental design criteria that should be considered prior to the development of a cement for use as a bone cement or in applications requiring a bone substitute. PMID- 15348505 TI - Encapsulation of apatite particles for improvement in bone regeneration. AB - The layering of fluorapatite on hydroxyapatite bodies provides a means of decreasing the solubility of hydroxyapatite, providing fluoride for possible stimulation of bone formation and delaying the release of calcium and phosphate from the more soluble hydroxyapatite. The purpose of this work was to encapsulate hydroxyapatite particles with fluorapatite spanning a thickness more than several crystallites deep. A three-step procedure was employed. Hydroxyapatite powder was immersed in an electrolyte solution until an equilibrium was established between the solid and the dissolved calcium at pH 4.67 and 37 degrees C. Equilibrium was determined by measurement of dissolved calcium with a calcium-specific ion specific electrode. A 5x10(-2) M ammonium fluoride added to the suspension resulted in a rapid decrease of both calcium and fluoride in the solution. Analysis with X-ray diffraction indicated that a fluoride rich layer containing calcium fluoride deposited onto the particle surface. Scanning electron microscopy revealed submicron spherical precipitate clusters on the hydroxyapatite particles. These clusters transformed to fluorapatite by soaking in a 0.1 M K(2)HPO(4) solution at pH 8 and 70 degrees C. A total time of 10 h was necessary for complete transformation of CaF(2) into fluorapatite. PMID- 15348506 TI - Albumin adsorption on cibacron blue F3G-A immobilized onto oligo(ethylene glycol) terminated self-assembled monolayers. AB - Self-assembled monolayers can be tailored with specific ligands to a certain protein and at the same time prevent the non-specific adsorption of other proteins. Cibacron Blue F3G-A (CB-thiol) was successfully immobilized onto tetra(ethylene glycol)-terminated alkanethiol (CB-thiol). The affinity of human serum albumin (HSA) to immobilized Cibacron Blue F3G-A was studied using mixed thiolate self-assembled monolayers on gold with different n-alkyl chain lengths and functional terminal groups (CH(3)-; OH- and tetra(ethylene glycol)). Surfaces were characterized using X-ray photoelectron spectroscopy and water contact angle measurements. Albumin adsorption and exchangeability of the adsorbed albumin molecules with other albumin molecules in solution were evaluated using (125)I radiolabeled HSA. Competitive adsorption between albumin and fibrinogen to the different self-assembled monolayers (SAMs) was also investigated. Results showed that the incorporation of CB-thiol on the monolayers does not increase the HSA adsorption and reversibility on the SAMs. However, although specific adsorption of HSA to the immobilized Cibacron Blue F3G-A was not demonstrated, the presence of CB-thiol decreases the affinity of fibrinogen to the OH-terminated SAMs. PMID- 15348507 TI - Free volume and mechanical properties of Palacos R bone cement. AB - The free volume and the mechanical properties of Palacos R bone cement were determined from positron annihilation lifetime spectroscopy (PALS) and from dynamic mechanical thermal analysis (DMA) in the temperature ranges 24-220 degrees C and 30-120 degrees C, respectively. The heating of bone cement caused an appreciable reduction of the free volume, measured as a decrease in the ortho positronium lifetime tau(3) from 2.04 to 1.91 ns, as well as a clear increase in the storage modulus from 2.3 to 3.0 GPa. The changes in free volume and storage modulus after the heat treatment were interpreted as an effect of elimination of residual monomer from the bone cement. The free volume of the bulk-polymerized phase of bone cement was estimated from a simple difference method, suggesting that the residual monomer was eliminated from the bone cement between 60 and 90 degrees C, thus implying a glass transition temperature of only 60 degrees C for the bulk-polymerized phase. The spherical free volume cavity size estimated from the ortho-positronium lifetime V(tau(3)), and the storage modulus E'(storage) from DMA measurements were found to correlate by a linear relationship throughout the studied temperature range, and the correlation appeared to be independent of the presence of residual monomer. PMID- 15348508 TI - Effects of baicalin-modified poly(D,L-lactic acid) surface on the behavior of osteoblasts. AB - In the present study, the functions of rat calvaria osteoblasts on baicalin modified poly(D,L-lactic acid) (PDLLA) films were investigated in vitro. The surface characteristics of surfaces (both modified and control) were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). Cell morphologies on these surfaces were examined by scanning electron microscopy (SEM). Cell adhesion and proliferation were used to assess cell growth on the modified and control surfaces. The MTT assay was used to determine cell viability and alkaline phosphatase (ALP) activity was performed to evaluate differentiated cell function. Compared to control films, cell attachment of osteoblasts on baicalin-modified PDLLA film was significantly higher (P<0.05 and P<0.01) after 6 h and 8 h culture, and cell proliferation was also significantly greater (P<0.05 and P<0.01) at the end of 4th and 7th day, respectively. The MTT assay suggested that the cell viability of osteoblasts cultured on baicalin-modified PDLLA film was significantly higher (P<0.05) than that seeded on the control. Meanwhile, the ALP activity of osteoblasts cultured on modified films was also considerably enhanced (P<0.01) compared to that found on control. These results revealed that the biocompatibility PDLLA could be improved by surface modification with baicalin. PMID- 15348509 TI - Multi-technique characterization of retrieved bone cement from revised total hip arthroplasties. AB - The purpose of this study was to assess the chemical composition, structure and degree of double bond conversion of retrieved bone cement from 29 total hip replacement revision arthroplasties, employing a multi-technique approach. Scanning electron microscopy revealed a porous cement surface, which replicated the characteristics of bone or femoral stem surface irregularities. Fourier transform infrared spectroscopy indicated that the retrieved bone cement samples were covered by a well-organized proteinaceous film rich in amides and alcohols, probably because of the adsorption of species from body tissues and fluids. X-ray fluorescence spectrometry showed the presence of potassium, sodium, calcium and phosphorus, implying the development of a mineralization process of the adsorbed biofilm. X-ray microtomography demonstrated a dense porous network in the bulk material comprised of macropores with a mean diameter >1 mm. FTIR analysis of the degree of double bond conversion of retrieved samples was in the order of 70%, similar to that of samples prepared in vitro in air, but 30% lower relative to their counterparts mixed in vitro and set in water. The effect of the adsorption of species onto bone cement surface on the reactivity of the material with the surrounding tissues and materials, is currently unknown. The results of this investigation reveal that the in vivo aging pattern of bone cements may involve alterations, which cannot be simulated under current in vitro protocols, emphasizing the necessity for adopting in vivo approaches including retrieval studies in assessing bone cement properties. PMID- 15348510 TI - Surface modification of a porous hydroxyapatite to promote bonded polymer coatings. AB - Porous hydroxyapatite (Hap) blocks were sintered at several temperatures and methyl methacrylate (MMA) grafted onto the surface in a 2-step heterogeneous system as a model example for surface modification. First, sintered porous Hap was modified with 2-methacryloyloxyethylene isocyanate (MOI) monomer in anhydrous dimethyl sulfoxide using di-n-butyltin (IV) dilaurate as a catalyst and hydroquinone as an inhibitor. Amount of the introduction of MOI monomer on porous Hap was 1.62 wt % at sintered temperature 800 degrees C, 0.68 wt % at it of 1000 degrees C, and 0.59 wt % at it of 1200 degrees C. Scanning electron microscopy (SEM) showed that porous Hap pore size and shape before and after MOI treatment were unchanged. Second, graft polymerization with MMA through the vinyl bond on porous Hap was conducted using alpha,alpha'-azobis isobutyronitrile (AIBN) as an initiator. Amount of Grafted PMMA on the MOI modified porous Hap was 2.84 wt % at sintered temperature of 800 degrees C, 6.97 wt % at it of 1000 degrees C, and 6.27 wt % at it of 1200 degrees C. MOI-modified and PMMA-grafted porous Hap were characterized using Fourier transform infrared (FT-IR) spectroscopy. The compressive strength of sintered porous Hap with grafted PMMA increased about 2.7 6.7 times compared to intact porous Hap. This 2-step surface modification on porous Hap is widely applicable to graft polymerization with vinyl polymer and conjugation with a protein or an oligopeptide, such as growth factor or an adhesion molecule, to improve Hap mechanical properties and functionality. PMID- 15348511 TI - Age-related changes in cyclic voltammetry and potentiodynamic studies of normal human dentine. AB - Impedance spectroscopy is one of the non-destructive techniques used by researchers to measure electrical resistance of biological tissues and ceramics. The purpose of this study is to investigate the voltage-current (V-I) characteristics of sound human dentine from young and old teeth, using cyclic voltammetry and potentiodynamic techniques. Dentine samples were prepared from freshly extracted sound third molars. After electrical measurements, dentine samples were characterized using scanning electron microscopy (SEM). Cyclic voltammetric measurements showed that variation of current through sample as a function of applied voltage is linear for dry samples of both age groups. However, for wet samples V-I characteristic were found to be different. The resistivity of dry young dentine is greater than that of old dentine in dry environment, whereas, it was found to be opposite for wet dentine samples. Using the same voltage sweep in potentiodynamic measurements dry samples display similar traces to controls suggesting that the dry dentine acts as an insulator. The number of dentinal tubules and their diameter has been found to decrease with increasing age. We propose that these changes determine the changes in electrical characteristics of sound human dentine. In spite of increasing use of electrical techniques to understand electrical properties of teeth, it is clear from this study that local structural variations and environment have a marked influence. Therefore, this baseline data needs to be considered in any future study or clinical application. PMID- 15348512 TI - Crystallization modifies osteoconductivity in an apatite-mullite glass-ceramic. AB - The response to implantation of novel apatite glass-ceramics was evaluated using a weight bearing in vivo bone implant model. Five novel glasses with varying calcium to phosphate ratios were cast as short rods and heat-treated to crystallize principally apatite. One glass ceramic had an apatite stoichiometry (Ca : P=1.67); three were phosphate-rich and one calcium-rich. One of the phosphate-rich glasses was also tested in its glassy state to determine the effect of crystallization on the biological response. Rods were implanted into the midshaft of rat femurs and left for 28 days. The femurs were then harvested and processed for scanning electron microscopy, energy dispersive X-ray microanalysis and conventional histology as ground and polished sections. Four of the materials exhibited evidence of osseointegration and osteoconduction. However, there was a marked inflammatory response to one of the phosphate-rich glass-ceramics, and to the non-crystallized glass. Crystallization of the latter significantly improved the bone tissue response. The glass-ceramic with an apatite stoichiometry elicited the most favorable response and merited further study as an osteoconductive bone substitute in maxillofacial and orthopedic surgery. PMID- 15348513 TI - Adhesion of bone cells to ion-implanted titanium. AB - The use of ion-implantation to encourage osseointegration has been investigated using an in vitro model cell culture system and surface analysis. Polished titanium discs were implanted with calcium, potassium and argon ions. The adhesion of bone-derived cells was measured using radioactively labeled cells and the morphology examined using scanning electron microscopy. Similar numbers of cells were found to adhere to the potassium and argon-implanted titanium as to control (non-implanted) titanium. However, adhesion to the calcium-implanted titanium discs was significantly reduced. Moreover, although the cells were found to be well spread on the calcium and potassium-implanted titanium, a much greater proportion of cells appeared to remain rounded and poorly attached on the argon implanted surface. These differences are discussed in relation to the observed surface roughness and chemistry, which were assessed using interferometry and X ray photoelectron spectroscopy, respectively. PMID- 15348514 TI - Effect of resin application time on bond strength of polymer substrate repaired with particulate filler composite. AB - Four different polymeric substrates were treated with unfilled intermediate resin (IMR) for various lengths of time (15 s, 3 min, 1 h, 24 h, and 2 weeks) to find out the effect of IMR treating time on shear bond strength between the substrate and a particulate filler composite. The IMR used in the study was a diacrylate monomer resin and the particulate filler composite was added on it. Two of the tested substrates were fiber-reinforced composites (FRCs). Two other substrates were cross-linked bisphenol-A-glycidyl dimethacrylate/triethyleneglycol dimethacrylate (BisGMA/TEGDMA) substrate and linear polymethylmethacrylate substrate. The bond strength of the particulate filler composite to the BisGMA/TEGDMA substrate was significantly higher compared to the other substrates (p<0.001). A correlation between increasing shear bond strength and longer treating time were found with the BisGMA/TEGDMA substrates (r=0.594, p<0.001) whereas no correlation was found with the other substrates. Prolonging of IMR treating time enhanced the BisGMA/TEGDMA substrate bond strength to the particulate filler composite but it had no clear effect on the bond strength of other substrates. PMID- 15348515 TI - Characterization of PC12 cell proliferation and differentiation-stimulated by ECM adhesion proteins and neurotrophic factors. AB - Among the various elements which influence axonal outgrowth in vivo is the physicochemical interaction of actively outgrowing nerve fibers with the various substrata they encounter during differentiation. Several experiments have explored the role of the extracellular matrix (ECM) in the control of neuronal differentiation. The nature, however, of the interactions between neurons and components of the ECM during regeneration and development are largely a matter of speculation. Although previous studies have already explored the influence of a number of ECM adhesion proteins and neurotrophic factors on neurite outgrowth, none have been carried in a systematic approach that allows for the simultaneous comparison of different surface conditions in relation to different neurotrophic factors. Motivated by the necessity of establishing controlled environments that allow for the rational design of stable neuronal/biomaterial interfaces, the long term effects of NGF and FGF-2 on the behavior of PC12 cells plated on collagen and laminin modified surfaces were evaluated. A pheochromocytoma cell line derived from transplantable rat adrenal medulla, PC12 cells have been commonly employed as an instructive model for studying the underlying mechanisms of neuronal differentiation. Long-term characterization of PC12 proliferation and neuronal differentiation for an experimental duration of 7-22 days was achieved by both qualitatively and quantitatively assaying for cell count, neurite number, neurite mean length, and neurite stability. Neurite stability was determined in terms of resistance to loss after neurotrophic factor (NGF/FGF-2) withdrawal. The present findings demonstrate that ECM adhesion proteins collagen and laminin are equally effective in promoting PC12 proliferation. It was noted as well that NGF supplemented collagen cultures are significantly more efficient in providing long term support to PC12 differentiation in terms of neurite number, mean length, and stability. PMID- 15348516 TI - Fracture toughness of tooth acrylics. AB - The hardness, fracture toughness, toughness, flexural strength and Young's moduli of three acrylic tooth polymers were investigated. The first polymer was based on a conventional homopolymer poly(methylmethacrylate). The second was based on cross-linked poly(methylmethacrylate) with an uncross-linked poly(methylmethacrylate) coating. The third material was based on an interpenetrating polymer network (IPN) of a cross-linked and uncross-linked poly(methylmethacrylate). All three polymers had similar hardness values. The cross-linked and IPN polymers had higher fracture toughness (K(IC)) and toughness (G(IC)) values than the conventional homopolymer poly(methylmethacrylate) polymer and lower flexural strengths (sigma(f)). The toughness of the cross-linked and IPN polymers was higher due to crack deflection around the polymer bead structure and the polymer beads acting as crack pinning sites. PMID- 15348518 TI - Chlorhexidine-containing mucoadhesive polymeric compacts designed for use in the oral cavity: an examination of their physical properties, in vitro/in vivo drug release properties and clinical acceptability. AB - This study describes the formulation, physicochemical and mucoadhesive properties and in in vitro/in vivo release of chlorhexidine from mucoadhesive, polymeric compacts, designed for application within the oral cavity. Compacts were prepared by direct compression of mixtures containing 100 mg sodium carboxymethylcellulose (NaCMC), 25 mg hydroxyethylcellulose (HEC)/75 mg polyacrylic acid (PAA) and 75 mg HEC/25 mg PAA. The mechanical and mucoadhesive properties of the drug-loaded compacts were examined using a texture analyzer in compression and tension modes, respectively. Evaluation of mucoadhesion was performed using a mucin-coated gauze substrate. In vitro release of chlorhexidine was performed under sink conditions (phosphate buffered saline, pH 7.0, 37 degrees C) using a Caleva 7ST dissolution apparatus. Salivary chlorhexidine levels were determined following intra-oral placement of drug-containing formulations. Quantification of the mass of chlorhexidine released both in vitro and in vivo was performed using HPLC with ultraviolet detection. Furthermore, the in vivo acceptability of the various polymeric compacts was assessed in volunteers using standard questionnaires. Compacts composed of HEC/PAA exhibited greater in vivo retention than those composed of NaCMC. Compacts composed of 25 mg PAA and 75 mg HEC displayed greatest patient acceptability. Introduction of chlorhexidine into these compacts did not significantly compromise either the work required for compact fracture or the in vitro mucoadhesion. Controlled release of chlorhexidine from these compacts was observed both in vitro and in vivo, the concentration of chlorhexidine in saliva exceeding the minimum inhibitory concentration of the common oral pathogens over the study period. In light of the patient acceptability and in vivo performance, it is suggested that the compact composed of 25 mg PAA/75 mg HEC containing 10 mg chlorhexidine offers considerable promise for use as an antimicrobial agent in the oral cavity. PMID- 15348519 TI - Non cell adhesive photopolymerized cross-linked layers (I): synthesis and characterization. AB - The aim of this study is to design an effective hydrogel, which has good potential for application in the field of tissue regeneration. Ultimately, the polymeric biomaterial will be used as a bottom, non-cell adhering layer of a polymeric scaffold, which will inhibit the adhesion and proliferation of fibroblast cells. A series of terpolymer hydrogels were prepared from MMA/HPOEM/MePOEM monomers based on varying formulations. The gels were polymerized via free radical polymerization under UV and characterized by TGA, DSC and FTIR studies. Furthermore, the double bond conversion, gel content and swelling properties of the polymer were analyzed and correlated to its crosslinking density, which revealed details about the hydrogel's character. Finally, cell culture experiments were used to compare the UV-cured polymer to its thermally derived non-functionalized polymer of the same formulation. Inverted optical microscopy was used to demonstrate cell-resistance of both materials with respect to a control Tissue Culture PolyStyrene plate. PMID- 15348520 TI - Tensile force testing of optimized coin-shaped titanium implant attachment kinetics in the rabbit tibiae. AB - In the present study, the bone response of titanium implants at early bone healing stages, was evaluated using a tensile test. Test surface of coin-shaped cp. titanium implants were standardized by grit blasting with TiO(2), grain size 180-220 microm. The surface topography of the implant specimens was examined by SEM, and by a confocal laser scanner for evaluation of S(a), S(t) and S(dr). The implants were placed onto the leveled site on the tibia of 12 New Zealand White rabbits, 4 implants in each animal. The rabbits were divided into three groups with different observation times i.e. 2, 4 and 6 weeks. The retention of 12 implants were tested by measuring the pull-out force needed to detach the implant from the bone. There was a significant increase in implant retention from 2 to 4 and to 6 weeks healing time (p<0.05). Four implants from each time point were randomly chosen for histological evaluation. The histological appearance of the implant-bone interface at the different healing times showed noticeable differences in the degree of bone healing and maturation, suggesting that, in rabbits, 6 weeks healing time is a suitable observation point for tensile testing of surface optimized osseointegrating implants. PMID- 15348521 TI - Radiation enhanced modification of HDPE for medical applications. AB - Comparison of gamma irradiation induced change in properties in terms of thermal and mechanical properties between two grades of HDPE and UHMWPE was carried out. It was found that the responses to irradiation of two grades of HDPE investigated were close whereas a difference in response was found between HDPE and UHMWPE. The irradiation dose that caused the lowest wear and highest hardness for UHMWPE was 500 kGy. When irradiation dose was above 500 kGy, no significant changes in wear and hardness properties were observed. The irradiation dose for HDPE, both 2208J and 7000F, that caused the wear resistance and hardness comparable to irradiated UHMWPE at 500 kGy was 1000 kGy. In addition, the dose of 750 kGy was needed for HDPE to achieve the similar level of wear resistance as non-irradiated UHMWPE. PMID- 15348522 TI - Multi-axial fatigue failure of orthopedic bone cement - experiments with tubular specimens. AB - Bone cement is subjected to multi-axial cyclic loading when used to fixate orthopedic prostheses for joint arthroplasty. In this study, tubular specimens of poly(methylmethacrylate) (PMMA) bone cement are subjected to internal pressure and cyclic axial loading to ascertain the influence of multi-axial loading on fatigue life. As expected, it was found that the probability of survival of specimens under multi-axial loading was very much reduced relative to specimens loaded uniaxially. Furthermore, the variability of the fatigue life was increased by multi-axial loading. In conclusion, the results point to the importance of characterizing the behavior of bone cements under the multi-axial fatigue experienced in vivo, and of the importance of accounting for the multi-axial stress state when predicting implant longevity. PMID- 15348523 TI - Surface crystalline phases and nanoindentation hardness of explanted zirconia femoral heads. AB - One new and nine explanted zirconia femoral heads were studied using glancing angle X-ray diffraction, scanning electron microscopy, and nanoindentation hardness techniques. All starting zirconia implants consisted only of tetragonal zirconia polycrystals (TZP). For comparison, one explanted alumina femoral head was also studied. Evidence for a surface tetragonal-to-monoclinic zirconia phase transformation was observed in some implants, the extent of which was varied for different in-service conditions. A strong correlation was found between increasing transformation to the monoclinic phase and decreasing surface hardness. Microscopic investigations of some of the explanted femoral heads revealed ultra high molecular weight polyethylene and metallic transfer wear debris. PMID- 15348524 TI - The effect of saliva on surface hardness and water sorption of glass-ionomers and "compomers". AB - A study is reported in which commercial dental materials (glass-ionomers, resin modified glass-ionomer and polyacid-modified composite resins) in the form of discs of dimensions 6 mm diameter x 1 mm thickness were prepared and exposed to natural salivas (parotid and unstimulated whole), artificial saliva and water for up to 1 year. Surface hardness was measured at various time intervals, and water sorption characteristics were determined. For all types of material, storage in artificial saliva gave specimens of lowest surface hardness by amounts that were generally significant to p<0.05, whereas no differences were found between specimens stored in water or either of the natural salivas. Water sorption characteristics were found to be unaffected by the nature of the storage medium. These results contrast with some previous findings and were not expected, given the known surface reactions between salivas and glass-ionomers, or the known enzymic degradation of composite resins. They demonstrate, however, that the current widespread practise of employing pure water for storage of specimens in laboratory studies is acceptable. PMID- 15348525 TI - Synthesis and characterization of nano-biomaterials with potential osteological applications. AB - The manufacture of high-surface area, un-agglomerated nano-sized (1-100 nm) bioceramic particles are of interest for many applications including injectable/controlled setting bone cements, high strength porous/non-porous synthetic bone grafts, and the reinforcing phase in nano-composites that attempt to mimic the complex structure and superior mechanical properties of bone. In the present study, we report on the manufacture of nano-particle hydroxyapatite powders by several wet chemical methods, which incorporate a freeze-drying step. In particular, it was found that the emulsion-based syntheses yielded powders with high surface areas and small primary particle sizes. Freeze drying rather than oven drying of powders prepared by conventional wet chemical synthesis yielded a nano-sized powder with a comparatively higher surface area of 113 m(2)/g. All powders were calcined in air in a furnace at 900 degrees C to investigate the effects of synthesis method on phase purity and surface area. The materials were characterized by a range of analytical methods including Fourier transform infrared spectroscopy employing the photo acoustic (PAS-FTIR) sampling technique, BET surface area analysis, X-ray powder diffraction (XRD), and the particles were examined using a transmission electron microscope (TEM). PMID- 15348526 TI - Microstructure analysis of calcium phosphate formed in tendon. AB - The surface of soft tendon tissue has been modified using calcium phosphate in order for the tendon to directly connect with hard bone and reconstruct an injured ligament. Calcium phosphate was coated onto the tendon in a soaking process using alternating a CaCl(2) (200 mM) and a Na(2)HPO(4) (120 mM) solution. According to SEM/EDX observations, calcium phosphate was formed, not only on the tendon surface, but also inside the tendon tissue. When the tendon was treated with seven soaking cycles, calcium phosphate was detected between 0-500 microm from the tendon surface. According to TEM observations, the crystal morphology of calcium phosphate depends on the distance from the surface. Hydroxyapatite crystals were observed near the surface, while octa-calcium phosphate crystals could be observed further from the surface, thus at initial soaking. The crystals were formed on collagen fibrils in spaces between the collagen fibrils with the c axes of the crystals aligned parallel with the collagen fibrils. This finding suggests Ca(2+) ions to interact with the tendon surface, most probably with the carboxyl functional groups of collagen, and subsequently forming nucleation centers for the crystals. PMID- 15348527 TI - Effect of glass-ceramic microstructure on its in vitro bioactivity. AB - Two routes were used to obtain a glass-ceramic composed of 43.5 wt % SiO(2) - 43.5 wt % CaO - 13 wt % ZrO(2). Heat treatment of a glass monolith produced a glass-ceramic (WZ1) containing wollastonite-2M and tetragonal zirconia as crystalline phases. The WZ1 did not display bioactivity in vitro. Ceramizing the glass via powder technology routes formed a bioactive glass-ceramic (WZ2). The two glass-ceramics, WZ1 and WZ2, were composed of the same crystalline phases, but differed in microstructure. The in vitro studies carried out on WZ2 showed the formation of an apatite-like layer on its surface during exposure to a simulated body fluid. This paper examined the influence of both chemical and morphological factors on the in vitro bioactivitity. The interfacial reaction product was examined by scanning and transmission electron microscopy. Both instruments were fitted with energy-dispersive X-ray analyzers. Measurements of the pH made directly at the interface of the two glass-ceramics were important in understanding their different behavior during exposure to the same physiological environment. PMID- 15348528 TI - Enhanced performance of fluorine substituted hydroxyapatite composites for hard tissue engineering. AB - Dense fluorine (F) substituted hydroxyapatite composites with yttria-doped zirconia (Y-TZP) and/or alumina (Al(2)O(3)) were successfully fabricated without applying pressure at 1400 degrees C for 3 h. The suppression of decomposition via the formation of a fluor-hydroxyapatite (FHA) solid solution allowed the sintered body to reach full density. Such fully densified FHA-composites exhibited improved mechanical properties, such as strength, toughness, and hardness, having values of more than approximately 2-4 times higher than those of pure HA or HA composites. The proliferation behavior of osteoblast-like cells on the FHA composites showed no cytotoxicity and comparable cell viability to that observed in pure HA for up to 10 days. PMID- 15348529 TI - Blood response to plasticized poly(vinyl chloride): dependence of fibrinogen adsorption on plasticizer selection and surface plasticizer level. AB - The high level of plasticizer in plasticized poly(vinyl chloride) (PVC) ensures that plasticizer selection has an important influence on the suitability of PVC to function in blood-contacting applications. In this study, three types of plasticized PVC in sheet form, with di-(2-ethylhexyl)phthalate (DEHP), tri-(2 ethylhexyl)trimellitate (TEHTM) and n-butyryltri-n-hexyl citrate (BTHC) as plasticizer, were selected for assessment and single solute fibrinogen adsorption was utilized as an initial index of interactions with blood components. Fibrinogen adsorption behavior shows a strong dependence on the plasticizer selection, plasticizer level at the surface and the adsorption conditions, such as adsorption time and fibrinogen solution concentration. Results indicate that BTHC plasticized PVC possesses the lowest adsorption capacity in the three types of plasticized PVC, while TEHTM plasticized PVC seems to have the strongest reactivity in certain fibrinogen solution concentrations. The alteration of surface plasticizer level was achieved by a methanol-cleaning treatment with a variety of cleaning times and the fibrinogen adsorption on plasticized PVC decreases with the reduction of surface plasticizer level. The migration behavior of two phthalate esters (DEHP and TEHTM) was evaluated using UV-Spectrophotometer to determine the plasticizer level at the surfaces. In addition, the fibrinogen adsorption mechanism was examined with Freundlich adsorption modeling. PMID- 15348530 TI - Tissue reactions of subcutaneously implanted mixture of epsilon-caprolactone lactide copolymer and tricalcium phosphate. An electron microscopic evaluation in sheep. AB - Biodegradable polymers, mainly derivates of alpha-hydroxy acids, are widely used today in oral- and maxillofacial surgery, orthopedics, and other fields of surgery. These biomaterials are well tolerated by living tissue and fracture fixation devices made of polylactic or polyglycolic acid are clinically widely used today. Still, there are some problems in application of biodegradable polymers. Abacterial inflammatory reactions have been noticed after the clinical introduction of these devices. Both swelling and pain at the site of implantation have also been reported. The etiology of this inflammatory reaction is still unknown, despite the numerous studies. Therefore, the aim of the present study was to further characterize this inflammatory reaction in detail, by electronmicroscopy. We prepared a mixture of epsilon-caprolactone-lactide copolymer and tricalcium phosphate and placed it in the dermis in 12 sheep. Follow-up times were 9, 14, 24, and 52 weeks. We found that the mixture caused a mild inflammatory reaction. There were no signs of cell damage. Fibroblasts, macrophages, and eosinophils were found adjacent to the copolymer. The mixture is easy to handle and can be moulded into different shapes in room temperature. The results encourage us to continue our studies to develop a filling material for bone defects. PMID- 15348531 TI - Formation of highly oriented hydroxyapatite in hydroxyapatite/titanium composite coating by radio-frequency thermal plasma spraying. AB - Highly oriented hydroxyapatite (HA) coatings with excellent adhesion were successfully obtained on titanium (Ti) and titanium alloy through a radio frequency thermal plasma spraying method. The ratio of HA and Ti powders supplied into the plasma was precisely controlled by two microfeeders so as to change the composition from Ti-rich to HA-rich toward the upper layer of the formed coatings. The bond (tensile) strength of the HA/Ti composite coatings was ca. 40 50 MPa. XRD patterns showed that the topmost HA layer of the coatings had an apatite structure with (00l) preferred orientation. The degree of this orientation showed a tendency to increase with an increase in the substrate temperature during spraying. PMID- 15348532 TI - Effect of chromium content on structure and mechanical properties of Ti-7.5Mo-xCr alloys. AB - The present work is a study of a series of Ti-7.5Mo-xCr alloys with the focus on the effect of chromium content on the structure and mechanical properties of the alloys. Experimental results show that low hardness, strength and modulus binary Ti-7.5Mo alloy is comprised primarily of fine, acicular martensitic alpha' phase. When 1 wt % Cr is added, a small amount of beta phase is retained. With 2 wt % or more chromium added, the entire alloy becomes equi-axed beta phase with bcc crystal structure. The average beta grain size decreases with Cr content. When the alloy contains about 2-4 wt % Cr, a metastable omega phase is present. In Ti 7.5Mo-2Cr alloy appears the highest omega intensity accompanied with high microhardness, bending strength and modulus. The omega-induced embrittling effect is most profound in Ti-7.5Mo-2Cr alloy that exhibits a terrace type fracture surface covered with numerous micron-sized dimples. The alloys with higher Cr contents show normal ductile type fractography with much larger deformation dimples. The present results indicate that Ti-7.5Mo-(4-6)Cr alloys seem to be potential candidates for implant application. PMID- 15348533 TI - Thermal analysis of poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogels. AB - The influence of water on the physical properties of a hydrogel is important for understanding natural tissues and in designing synthetic materials to replace them. In this study, poly (2-hydroxyethyl methacrylate) (pHEMA) was used as a model system to understand how water interacts with the polymer of a hydrogel. Thermal analysis methods (thermogravimetric analysis coupled to mass spectrometry and differential scanning calorimetry) were used to determine: (i) the total water content of pHEMA gels; (ii) how this water was lost during heating; (iii) the relationship between water content of the gel and its glass transition temperature; and (iv) the behavior of the water in the gel on cooling. Previous researchers have invoked various models to describe the organization of water in a hydrogel. In this study, the simplest model which could explain all of the results from the different thermal analysis techniques was one which consisted of three classes of water: (i) hydration water in close proximity to the polymer; (ii) interstitial water in regions or cavities surrounded by polymer chains; and (iii) bulk water. PMID- 15348534 TI - Contact damage in model dental multilayers: an investigation of the influence of indenter size. AB - This paper presents a combined experimental and computational study of the influence of indenter ball size on contact damage in model multilayered structures with equivalent elastic properties to bonded dentin/crown structures. Following a brief description of restored tooth structures, prior work on the development of model dental multilayered structures is reviewed. The effects of indentation ball size are investigated within a combined experimental and computational/analytical framework. The observed cracking patterns at the onset of crack nucleation are shown to be associated with principal stress contours computed using finite element analysis. The implications of the results are discussed for the design of dental multilayers that are more resistant to crack nucleation and propagation. PMID- 15348535 TI - Preparation and characterization of novel silica-butyrylchitosan hybrid biomaterials. AB - Novel silica-butyrylchitosan hybrid biomaterials were produced by a sol-gel technique, using butyrylchitosan as the organic species incorporated into the silicon alkoxide (TEOS) based network. 3-acryloxypropyl trimethoxysilane (MPTMS) was used effectively to combine the organic and inorganic species to form uniform hybrid biomaterials. All the samples made were in the form of thin, flexible films with transparent clarity. The blood-clotting and platelet adhesion assay confirmed that these hybrid biomaterials displayed potential good blood compatibility. PMID- 15348536 TI - Indirect cytotoxicity evaluation of pseudowollastonite. AB - This study aimed to evaluate the cytotoxicity of substances leached by pseudowollastonite (CaSiO(3)). It has been previously shown that calcium (Ca(2+)) and silicate (SiO(3)(-)) ions are released from pseudowollastonite into biological solutions. Both of these ions are known to influence the biological metabolism of osteoblastic cells essential in the mineralization process and bone bonding mechanism. The indirect toxicity evaluation was performed by extraction method, according to International Standard Organization (ISO). Pseudowollastonite pellets obtained by solid-state reaction were incubated, in culture medium, during 24, 48, 72 or 168 h at different concentrations (5, 10, 15, 50, 100, 200 mg/ml). The cytotoxicity of each extract in presence of human osteoblastic cell line (SaOS-2) was quantitatively assessed by measuring the viability (succinate dehydrogenase activity, MTT), the membrane integrity (the uptake of the neutral red by viable cells, NR) as well as the cell necrosis by measuring the lactate dehydrogenase (LDH) released in the culture medium. No significant alteration of membrane integrity or cell suffering was detectable. However, increased cell metabolism was observed for cells exposed to pseudowollastonite extract with longest extraction time (168 h). In conclusion, mineral elements leached by pseudowollastonite do not significantly affect the metabolism of osteoblastic cells. PMID- 15348537 TI - Wettable phosphorylcholine-containing polymers useful in blood filtration. AB - The use of phosphorylcholine (PC)-containing polymers has long been acknowledged as a method to improve the haemocompatibility of blood contacting devices. Such polymers were investigated for coating leukocyte filters, as a means of preserving precious platelet numbers and function. It was demonstrated that by use of such coated filters, the platelet recovery could be significantly increased by some 30%. This was however, balanced by a decrease in the leukocyte removal efficiency of the filter. More problematic however, was the poor critical wetting surface tension (CWST) of the filter (45 mNm(-1)). This was due to the surface expression of the hydrophobic groups of the polymer in air. In order to obtain a filter that could be immediately wetted by the blood and hence remove the need for any detrimental priming of the filter, the PC polymer was modified in order to reduce its hydrophobic content. A series of new cross-linkable polymers was developed that contained other hydrophilic monomers, yet could film form to produce a coating stable to fluid contact. These materials demonstrated the required wetting characteristics (a CWST of >78 mNm(-1)), whilst retaining the excellent blood-contacting properties. PMID- 15348538 TI - Wear behavior of carbide coated Co-Cr-Mo implant alloy. AB - The wear behavior of a new type of metal carbide surface coating on Co-Cr-Mo implant alloy was studied. The coating was created using a microwave plasma assisted reaction. Codeposition of impurity diamond film, diamond particles, and soot was prevented by controlling process conditions. Wear tests were carried out using a sapphire ball-on-Co-Cr-Mo disc unidirectional sliding configuration with harsh conditions of high contact stress and slow sliding speed in both no lubrication, and deionized water lubrication environments. In the case of uncoated Co-Cr-Mo discs, the effect of deionized water lubrication was remarkable and reduced the wear factor by one order of magnitude compared to the no lubrication tests. The wear factor of carbide coated Co-Cr-Mo discs was slightly smaller than that of uncoated Co-Cr-Mo discs with deionized water lubrication (2.7 x 10(-6) mm3 N(-1) m(-1) vs. 4.2 x 10(-6) mm3 N(-1) m(-1)). The addition of deionized water lubrication did not greatly affect the wear factor of carbide coated Co-Cr-Mo discs. The influence of surface geometry resulting from the "brain coral-like" surface morphology of carbide layers on wear behavior was analyzed considering stress concentrations and effective contact area. PMID- 15348539 TI - Factors affecting the static shear strength of the prosthetic stem-bone cement interface. AB - Debonding of the stem-cement interface has been implicated in the initiation of failure of cemented femoral stems. The objective of this work was to examine some of the parameters which influence the interface static shear strength, including surface finish, cement type, pre-treatments and porosity. Surface finish was found to have the greatest effect on the interface strength. Increasing the surface roughness by a factor of 100 increases the interface shear strength by a factor of 20. However, increasing the surface roughness above a certain value was found to have no additional affect. This was due to failure in the cement itself rather than at the cement-stem interface. There were significant differences between some of the different cement types regarding the interface strength. Pre heating the stem produced a six fold reduction in cement porosity at the stem cement interface, however, resulting in only a minor influence on the static interface strength. Generally, no significant correlation was found between the cement porosity and the static interfacial shear strength. PMID- 15348540 TI - Impact behavior of hydroxyapatite reinforced polyethylene composites. AB - Hydroxyapatite particulate reinforced high density polyethylene composite (HA HDPE) has been developed as a bone replacement material. The impact behavior of the composites at 37 degrees C has been investigated using an instrumented falling weight impact testing machine. The fracture surfaces were examined using SEM and the fracture mechanisms are discussed. It was found that the fracture toughness of HA-HDPE composites increased with HDPE molecular weight, but decreased with increasing HA volume fraction. Examination of fracture surfaces revealed weak filler/matrix interfaces which can debond easily to enable crack initiation and propagation. Increasing HA volume fraction increases the interface area, and more cracks can form and develop, thus decreasing the impact resistance of the composites. Another important factor for the impact behavior of the composites is the matrix. At higher molecular weight, HDPE is able to sustain more plastic deformation and dissipates more impact energy, hence improving the impact property. PMID- 15348541 TI - The influence of complexing agent and proteins on the corrosion of stainless steels and their metal components. AB - The present work is devoted to the problem of biodegradation of orthopaedic implants manufactured from stainless steel. In vitro simulations of the biocompatibility of two types of stainless steel, AISI 304 and AISI 316L, and their individual metal components, i.e. iron, chromium, nickel and molybdenum, were carried out in simulated physiological solution (Hank's) containing complexing agents. Knowledge of the effects of the chemical and biological complexing agents, EDTA and proteins, respectively, on the corrosion resistance of a metal should provide a better understanding of the processes occurring in vivo on its surface. The behavior of stainless steels and metal components was studied under open circuit and under potentiostatic conditions. The concentration of dissolved corrosion products in the form of released ions was determined by differential pulse polarography (DPP) and atomic emission spectrometry using inductively coupled plasma (ICP-AES). The composition of solid corrosion products formed on the surface was analyzed by energy dispersive X-ray spectroscopy (EDS) and their morphology was viewed by scanning electron microscopy (SEM). The addition of EDTA and proteins to physiological solution increased the dissolution of pure metals and stainless steels. The effect of particular protein differs on different metals and alloys. PMID- 15348542 TI - The study of metal ion release and cytotoxicity in Co-Cr-Mo and Ti-Al-V alloy in total knee prosthesis - scanning electron microscopic observation. AB - We surgically retrieved two cobalt(Co)-chromium(Cr)-molybdenum(Mo) and five titanium(Ti)-aluminum(Al)-vanadium(V) alloy knee prostheses from patients because of mechanical failure and pain. We examined the distribution of the small particles which were released from the Co-Cr-Mo and Ti-Al-V alloys using a backscattered scanning electron microscopy (SEM). In addition we analyzed the metals in the artificial knee joints and the tissues adjacent to them using energy dispersive X-ray spectroscopy (EDS). We demonstrated that a myriad of fine particles, produced by the abrasion of both Co-Cr-Mo and Ti-Al-V alloys, accumulated in the synovial cells. As Co-Cr-Mo alloys disintegrate easily in the cells, Co dissolves from the peripheral areas of them, although Cr remains within the cells. In contrast Ti-Al-V alloys are very stable in the synovial cells. From these findings we conclude that the Co-Cr-Mo alloys are hazardous to the body as the alloys release Co which enters the body. In contrast the Ti-Al-V alloys are very stable and are patently safer. Artificial joints, however, are still in considerable need of improvement. PMID- 15348543 TI - Synthesis and characterization of poly(L-lactic acid) membranes: studies in vivo and in vitro. AB - The use of biodegradable polyesters as temporary structural supports in the recuperation of damaged live tissue is a promising area of research. Poly(L lactic acid) (PLLA) membranes can act as a support for cell fixation and growth or as a barrier against soft tissues invasion in recuperating bone tissues. In this work, five different types of PLLA membranes, which varied in their polymer solvent ratio and their content of plasticizer were studied. For the study in vivo, 6 mm diameter disks were inserted subcutaneously in the dorsal region of 15 Wistar rats, and the reactions on rats were studied 15 days later. In another series of experiments the samples were immersed in phosphate buffer, pH 7.4 at 37 degrees C, for 30 days. Membranes without plasticizer were morphologically dense and did not allow cell invasion nor tissue adherence, in contrast to membranes with plasticizer. While porosity enhanced cell fixation and growth, it made the membrane more fragile mechanically when compared to membranes without pores. PMID- 15348544 TI - The cell as a biomaterial. AB - For materials scientists, the cell is evidently a biomaterial - rich with polymers, surface forces, solvent-solute interactions, liquid-crystalline structures, etc. Yet, the language of the materials scientist is as foreign to the biological world as French is to Chinese. Little of the materials scientists' perspective has been brought to bear on the question of biological function. This review aims to begin bridging the gap between the two disciplines-to show that a materials-oriented approach has power to bring fresh insights into an otherwise impenetrably complex maze. In this approach the cell is treated as a polymer gel. If the cell is a gel, then a logical approach to the understanding of cell function is through an understanding of gel function. Great strides have been made recently in understanding the principles of polymer-gel dynamics. It has become clear that a central mechanism is the phase-transition - a major structural change prompted by a subtle change of environment. Phase-transitions are capable of doing work and such work could be responsible for much of the work of the cell. Here, we pursue this approach. We set up a polymer-gel-based foundation for cell behavior, and explore the extent to which this foundation explains how the cell achieves its everyday tasks. PMID- 15348545 TI - The effects of displacement rate and proteoglycan digestion on the fracture resistance of tissue grown from chondrocyte culture. AB - Fracture toughness of cartilage and cartilage replacement tissues is important in injury and disease. For example, cartilage is thought to weaken before it fibrillates in the disease osteoarthritis. Since both loading rate and proteoglycan content affect viscoelastic properties, they may both affect fracture toughness of cartilage and cartilage analogs. In this study, fracture toughness of tissue grown in chondrocyte culture was measured as a function of loading rate and proteoglycan digestion. Control tissue and tissue digested with chondroitinase ABC (cABC) to remove proteoglycans were tested at displacement rates of 0.1 and 0.5 mm/sec. Displacement rate had no effect on fracture toughness for either control or digested tissue. Proteoglycan digestion reduced tissue thickness by 30% and when evaluated on a material basis increased fracture toughness. There was no interaction between digestion and loading rate. When the fracture toughness was normalized to collagen content, which removed the effect of tissue shrinkage, there was no effect of proteoglycan digestion on fracture toughness. These data suggest that proteoglycans do not contribute to tissue toughness, other than by reducing thickness and increasing collagen density. PMID- 15348546 TI - Testing bone substitutes in a small animal model of revision arthroplasty. AB - This study evaluated a modification of the rat-pin model to enable testing of bone substitute materials. The model was characterized using the ceramic, beta tricalcium phosphate (betaTCP) as a filler. A 1 mm wide, 3.6 mm deep defect was created around a stainless steel (SS) implant in the proximal tibia of a rat. This defect was filled with a ceramic powder. Large particles (90-312microm) of betaTCP were mixed with Gelfoam to form a paste which was then molded around the proximal end of either an uncoated SS pin or a pin coated with hydroxyapatite (HA). The pin with its ceramic collar was then implanted into the proximal tibia of 16 male Sprague Dawley rats. Two animals with coated implants and two with uncoated implants were sacrificed at 3, 6, 14 and 26 weeks. Longitudinal sections of each tibia were stained with toluidine blue and labeled for tartrate resistant acid phosphatase (TRAP). There was initial fibrous tissue interposition around the implants which was completely remodeled around the HA coated pins but which persisted in apposition to the SS pins. The remodeling process peaked at 3 weeks around the HA coated pins and at 6 weeks around the uncoated implants. There was little remodeling around either implant by 26 weeks. There was considerable residual betaTCP present which was well tolerated as the particles were often encased in bone. The model has several characteristics of revision arthroplasty and the results demonstrate the suitability of this model for testing bone substitutes. PMID- 15348547 TI - Surface-modified 3D scaffolds for tissue engineering. AB - The aim of this work was to use sol-gel processing to develop bioactive materials to serve as scaffolds for tissue engineering that will allow the incorporation and release of proteins to stimulate cell function and tissue growth. We obtained organofunctionalized silica with large content of amine and mercaptan groups (up to 25%). The developed method can allow the incorporation and delivery of proteins at a controlled rate. We also produced bioactive foams with binary SiO(2)-CaO and ternary SiO(2)-CaO-P(2)O(5) compositions. In order to enhance peptide-material surface properties, the bioactive foams were modified with amine and mercaptan groups. These materials exhibit a highly interconnected macroporous network and high surface area. These textural features together with the incorporation of organic functionally groups may enable them to be used as scaffolds for the engineering of soft tissue. PMID- 15348548 TI - Mechanical properties of biodegradable polymer sutures coated with bioactive glass. AB - Combining commercially available Polyglactin 910 (Vicryl) sutures with bioactive glass powder offers new possibilities for application of composite materials in tissue engineering. Commercial bioactive glass (45S5 Bioglass) powder was used to coat Vicryl sutures and the tensile strength of the sutures was tested before and after immersion in simulated body fluid (SBF) as a means to assess the effect of the bioactive glass coating on suture degradation. Different gauge lengths (126.6 and 111.6 mm) and strain rates (2.54, 11.4 and 25.4 mm/min) were tested. The tensile strength of composite sutures was slightly lower than that of as-received Vicryl sutures (404 MPa versus 463 MPa). However after 28 days immersion in SBF the residual tensile strength of the coated sutures was significantly higher, indicating a protective function of the Bioglass coating. The tensile strength results were similar for the different gauge lengths and strain rates investigated. A qualitative explanation for the effect of bioactive glass coating on polymer degradation is offered. PMID- 15348549 TI - Effect of lipase treatment on the biocompatibility of microbial polyhydroxyalkanoates. AB - Films made from microbial polyesters polyhydroxybutyrate (PHB) and poly(hydroxybutyrate-co-hydroxyhexanoate) (PHBHHx) were treated by lipases and NaOH solution. The change of the polyester biocompatibility was evaluated by inoculating mouse fibroblast cell line L929 on films of PHB, PHBHHx and their blends. Polylactic acid (PLA) was used as a control. It was found that untreated PHB and PLA films gave a poor support to the growth of L929 cells, viable cell density ranged from 0.1x10(4}) to 0.7x10(4) per ml only. While films of pure PHBHHx and PHB blended with PHBHHx showed improved biocompatibility, viable cell density observed increased from 9.6x10(2) to 6x10(4) on blended films of PHB/PHBHHx in ratios of 0.9/0.1 to 0/1, respectively. This result showed PHBHHx has a better biocompatibility compared with PHB. Films of PHB, PLA and the blends treated with lipases and 1 N NaOH, respectively, showed an improved ability to support cell growth. Biocompatibility of PHB was approximately the same as PLA after the treatment, while PHBHHx and its dominant blends showed improved biocompatibility compared with PLA. The sensitivity of the treatments was reduced when PHBHHx content increased in the PHB/PHBHHx blends. All three lipase treatments demonstrated more biocompatibility increase on all the films compared with the results of NaOH treatment. Scanning electron microscopy showed that PHB films changed its surface from multi-porous to rough non-porous after the lipase or NaOH treatment. While PHBHHx films showed little change after these treatments. The results showed that the polyester surface morphology played an important role in affecting cell attachment and growth on these materials. PMID- 15348550 TI - Bending property and phase transformation of Ti-Ni-Cu alloy dental castings for orthodontic application. AB - Bending property of Ti-Ni-Cu alloy castings was investigated in a three-point bending test for orthodontic application in relation to the phase transformation. The compositions of the alloys were Ti-50.8Ni and Ti-40.8Ni-10.0Cu (mol %), and four cross-sectional shapes of the specimens were selected. Heat treatment was performed at 713, 753 or 793 K for 1.8 ks. The bending load changed by the cross sectional size and shape mainly because of the difference in the moment of inertia of area, but the load-deflection relation did not differ proportionally in the unloading process. The difference between the load values in the loading and the unloading processes was relatively small for Ti-Ni-Cu alloy. With respect to the residual deflection, there was no significant difference between Ti-Ni and Ti-Ni-Cu alloys with the same treatment condition. The load values in the loading and the unloading processes decreased by each heat treatment for Ti-Ni alloy; however, the decrease in the load values for Ti-Ni-Cu alloy was not distinct. It is proved that Ti-Ni-Cu alloy castings produce effective orthodontic force as well as stable low residual deflection, which is likely to be caused by the high and sharp thermal peaks during phase transformation. PMID- 15348551 TI - Hyaluronic acid and silver sulfadiazine-impregnated polyurethane foams for wound dressing application. AB - Five different kinds of PU foam wound dressings were prepared to investigate their wound healing capability. They include (i) PU+silver sulfadiazine (AgSD), (ii) PU+alginate (Al), (iii) PU+Al+AgSD, (i.v.) PU+hyaluronic acid (HA), and (v) PU+HA+AgSD. Physical properties and in vitro behaviors of AgSD release and fibroblast adhesion on those dressings were evaluated. From the drug release and fibroblast adhesion studies, it was observed that PU foam impregnated with both HA and AgSD shows good drug release behavior and low adhesion of the cells. Furthermore, the HA and AgSD-containing PU foam showed excellent wound healing effect without any inflammation or yellow cluster. The wound size decreased around 77% after 1 week application of that foam dressing onto a rat skin defect. PMID- 15348552 TI - Adhesion and morphology of fibroblastic cells cultured on different polymeric biomaterials. AB - Cell adhesion is influenced by the physical and chemical characteristics of the materials used as substrate for cell culturing. In this work, we evaluated the influence of the morphological and chemical characteristics of different polymeric substrates on the adhesion and morphology of fibroblastic cells. Cell growth on poly (L-lactic acid) [PLLA] membranes and poly(2-hydroxy ethyl methacrylate) [polyHEMA], poly(2-hydroxy ethyl methacrylate)-cellulose acetate [polyHEMA-CA] and poly(2-hydroxy ethyl methacrylate)-poly(methyl methacrylate-co acrylic acid) [polyHEMA-poly(MMA-co-AA)] hydrogels of different densities and pore diameters was examined. Cells adhered preferentially to more negatively charged substrates, with polyHEMA hydrogels being more adhesive than the other substractes. The pores present in PLLA membranes did not interfere with adhesion, but the cells showed a distinctive morphology on each membrane. PMID- 15348553 TI - Effect of thermal cycling on whisker-reinforced dental resin composites. AB - The mechanical properties of dental resin composites need to be improved in order to extend their use to high stress-bearing applications such as crown and bridge restorations. Recent studies used single crystal ceramic whiskers to reinforce dental composites. The aim of this study was to investigate the effects of thermal cycling on whisker-reinforced composites. It was hypothesized that the whisker composites would not show a reduction in mechanical properties or the breakdown of whisker-resin interface after thermal cycling. Silicon carbide whiskers were mixed with silica particles, thermally fused, then silanized and incorporated into resin to make flexural specimens. The filler mass fraction ranged from 0% to 70%. The specimens were thermal cycled in 5 degrees C and 60 degrees C water baths, and then fractured in three-point bending to measure strength. Nano-indentation was used to measure modulus and hardness. No significant loss in composite strength, modulus and hardness was found after 10(5) thermal cycles (family confidence coefficient=0.95; Tukey's multiple comparison test). The strength of whisker composite increased with filler level up to 60%, then plateaued when filler level was further increased to 70%; the modulus and hardness increased monotonically with filler level. The strength and modulus of whisker composite at 70% filler level were significantly higher than the non-whisker controls both before and after thermal cycling. SEM revealed no separation at the whisker-matrix interfaces, and observed resin remnants on the pulled-out whiskers, indicating strong whisker-resin bonding even after 10(5) thermal cycles. In conclusion, novel dental resin composites containing silica fused whiskers possessed superior strength and modulus compared to non-whisker composites both before and after thermal cycling. The whisker-resin bonding appeared to be resistant to thermal cycling in water, so that no loss in composite strength or stiffness occurred after prolonged thermal cycling. PMID- 15348554 TI - Physicochemical and microscopical study of calcific deposits from natural and bioprosthetic heart valves. Comparison and implications for mineralization mechanism. AB - Natural and bioprosthetic heart valves suffer from calcification, despite their differences in etiology and tissue material. The mechanism of developing calcific deposits in valve tissue is still not elucidated. The calcific deposits developed on human natural and bioprosthetic heart valves have been investigated and compared by physicochemical studies and microscopy investigations and the results were correlated with possible mechanisms of mineral crystal growth. Deposits from 16 surgically excised calcified valves (seven natural aortic and nine bioprosthetic porcine aortic valves) were examined by chemical analysis, FTIR, XRD, and SEM-EDS. The Ca/P molar ratio of the deposits from bioprosthetic valves (1.52+/-0.06) was significantly lower compared to that of the natural valves (1.83+/-0.03) (p=0.05, 1-way ANOVA). SEM-EDS examination of the two types of valve deposits revealed the coexistence of large (>20 microm) and medium (5-20 microm) plate-like crystals as well as microcrystalline (<5 microm) calcium phosphate mineral formations. The results confirmed the hypothesis that the mineral salt of calcified valves is a mixture of calcium phosphate phases such as dicalcium phosphate dihydrate (DCPD), octacalcium phosphate (OCP) and hydroxyapatite (HAP). DCPD and OCP are suggested to be precursor phases transformed to HAP by hydrolysis. The lower value of the Ca/P molar ratio found in the bioprostheses, in comparison with that corresponding in natural valves, was ascribed to the higher content in these deposits in precursor phases DCPD and OCP which were subsequently transformed into HAP. On the basis of chemical composition of the deposits and their morphology it is suggested that crystal growth proceeds in both types of valves by the same mechanism (hydrolysis of precursor phases to HAP) in spite of their differences in etiology, material, and possible initiation pathways. PMID- 15348555 TI - Hydroxyapatite coating of titanium by biomimetic method. AB - The biomimetic method was used in order to deposit, on titanium substrates, an hydroxyapatite (HA) coating. The bioactive HA layer was obtained by using, in the first stage of the process, a glass having the composition 2.5CaO.2SiO(2) different from the one proposed for the application of the biomimetic method. This glass can be obtained via sol-gel, a method that allows one to obtain, easily, very pure products. The growth of HA crystals was confirmed by Fourier transform infrared, SEM, EDS and X-ray photoelectron spectroscopy (XPS) results. The experimental results suggest that, as reported in the literature for other supports, the silicate ions released from the glass in the first stage bind themselves to the titanium support. In particular, from XPS analysis it is evident that the titanium substrate is well covered by a calcium phosphate layer of the type of HA. PMID- 15348556 TI - Bone formation following implantation of fibrous calcium compounds (beta Ca(PO3)2, CaCO3(aragonite)) into bone marrow. AB - Bone formation around three types of fibrous calcium-containing crystals has been examined histologically using rats. The implanted materials are (i) calcium metaphosphate (beta-Ca(PO(3))(2)) fibers having aspect ratios of 15-80 with 2-20 microm in diameter, (ii) beta-Ca(PO(3))(2)) fibers surface-modified using dilute NaOH and (iii) calcium carbonate (CaCO(3); aragonite phase) whiskers having aspect ratios of 15-40 with 0.5-3 microm in diameter. Beta-Ca(PO(3))(2) fibers show a mechanically high strength with a low modulus of elasticity, and the surface-modified fibers have a thin layer consisting of a calcium orthophosphate phase. CaCO(3) whiskers were used for comparison reasons. The materials were implanted for 4, 8, and 12 weeks into bone defects created in the bone marrow of rat tibiae. Cancellous bone formation was observed around beta-Ca(PO3)2 fibers, the surface-modified fibers and CaCO(3) whiskers after implantation for 12, 4 and 4 weeks, respectively. CaCO(3) whiskers were scarcely observed after 12 weeks for resorbing. The calcium phosphate fibrous materials show combined advantages of mechanically high strength for toughening a matrix phase and biological activities; thus, these materials may prove to be useful for novel applications in the biomedical field. PMID- 15348557 TI - Pathogenesis and prevention of biomaterial centered infections. AB - One of the major drawbacks in the use of biomedical materials is the occurrence of biomaterials centered infections. After implantation, the host interacts with a biomaterial by forming a conditioning film on its surface and an immune reaction towards the foreign material. When microorganisms can reach the biomaterials surface they can adhere to it. Adhesion of microorganisms to an implant is mediated by their physico-chemical surface properties and the properties of the biomaterials surface itself. Subsequent surface growth of the microorganisms will lead to a mature biofilm and infection, which is difficult to eradicate by antibiotics. The purpose of this review is to give an overview of the mechanisms involved in biomaterials centered infection and the possible methods to prevent these infections. PMID- 15348558 TI - Factors affecting the mechanical and viscoelastic properties of acrylic bone cement. AB - The aim of this paper is to report a series of experiments investigating the factors that influence the viscoelastic properties of acrylic bone cement. The effects of the brand of cement, the length of time since mixing, temperature, the hydration of the cement, and the influence of fat and or blood in the environment on the creep and stress relaxation behavior of the cement have been studied in laboratory-prepared specimens in tension, compression and four point bending. Although there are significant differences in the viscoelastic behavior of some of the different brands of polymethylmethacrylate based cements, these differences are small by comparison with the major effects that can be exerted by the length of time since mixing and some environmental factors. These effects have important practical consequences, especially with regard to the ability of bench top and theoretical studies to predict reliably the mechanical and viscoelastic behavior of acrylic cement in vivo. PMID- 15348559 TI - H2O2 production by cells on titanium and polystyrene surfaces using an in vivo model of exudate and surface related cell function. AB - The determination of secreted levels of reactive oxygen species by implant adherent cells in vivo is required for understanding of the role(s) of such reactive oxygen species for the tissue response around medical devices. A model with subcutaneous implants of c.p. titanium (Ti) or polystyrene (PS) (cell culture grade) inserted on the back of rats were used. Implants and associated cells were retrieved and assayed after 1, 3, 5, 7, 14, 21 and 28 days. Morphological analysis of exudate cells showed that polymorphonuclear leukocytes (PMN) predominated after one day whereas macrophages were predominant after three days. The number of implant-adherent cells, as reflected by measurement of DNA, decreased with time. Ultrastructural observations showed that macrophages were predominant cells in contact with the implant surface. Measurement of hydrogen peroxide (H(2)O(2)) secretion by implant-adherent cells during 40 min incubation ex vivo revealed a constitutive generation of 40-400 pmol H(2)O(2)/microg DNA, depending on implantation time. Stimulation with protein kinase C agonist phorbol myristate acetate (PMA) caused an increased H(2)O(2) generation by adherent cells at early (up to five days) but not later (7-28 days) time periods. No major differences between Ti and PS were observed. Taken together, these findings show that Ti and PS implant-adherent cells secrete H(2)O(2) under in vivo conditions. Further, a reduced capacity to mount an enhanced H(2)O(2) secretion upon stimulation was demonstrated at late time periods. The role of this mediator for biocompatibility remains to be established. PMID- 15348560 TI - Time dependence of the mechanical properties of GICs in simulated physiological conditions. AB - The mechanical properties of glass-ionomer cements (GICs) have been satisfactory for dental applications and have shown their potential in orthopedic surgery. Because the physiological environment in orthopedics is different from dentistry by unavoidable contamination with blood and other fluids such as normal saline used during an operation, the determination of GICs for orthopedic applications should be performed in an appropriate environment. The properties of a novel resin-modified GIC, S430, for orthopedic applications were evaluated in simulated orthopedic conditions by an early exposure to and long-term storage in normal saline. An early exposure to normal saline caused 20-60% reduction of its compressive and flexural properties, whereas long-term storage in normal saline showed slight changes of its mechanical properties. The effects were probably due to the disturbance of the cross-linking formation in the acid-base reaction and also the reduction of electrostatic interactions of the cross-linking polymeric chain of hydroxyethyl methacrylate (HEMA) in resin-modified GIC. PMID- 15348561 TI - Fibrin-mediated endothelial cell adhesion to vascular biomaterials resists shear stress due to flow. AB - In vitro endothelial cell (EC) seeding onto biomaterials for blood-contacting applications can improve the blood compatibility of materials. Adhesive proteins adsorbed from serum that is supplemented with the culture medium intercede the initial cell adhesion and subsequent spreading on material surface during culture. Nevertheless, physical and chemical properties of vascular biomaterial surface fluctuate widely between materials resulting in dissimilarity in protein adsorption characteristics. Thus, a variation is expected in cell adhesion, growth and the ability of cell to resist shear stress when tissue engineering on to vascular biomaterials is attempted. This study was carried out with an objective to determine the significance of a matrix coating on cell adhesion and shear stress resistance when cells are cultured on materials such as polytetrafluoroethylene (PTFE, Teflon) and polyethyleneterephthalate (Dacron), ultra high molecular weight polyethylene (UHMWPE) and titanium (Ti), that are used for prosthetic devices. The study illustrates the distinction of EC attachment and proliferation between uncoated and matrix-coated surfaces. The cell attachment and proliferation on uncoated UHMWPE and titanium surfaces were not significantly different from matrix-coated surfaces. However, shear stress resistance of the cells grown on composite coated surfaces appeared superior compared to the cells grown on uncoated surface. On uncoated vascular graft materials, the cell adhesion was not supported by serum alone and proliferation was scanty as compared to matrix-coated surface. Therefore, coating of implant devices with a composite of adhesive proteins and growth factors can improve EC attachment and resistance of the cells to the forces of flow. PMID- 15348562 TI - Influence of test protocol in determining the blood response to model polymers. AB - A multi-parametric, multi-center evaluation of three polymers was performed measuring their response to blood contact. The purpose of this study was to pinpoint differences in tests performed for assessing "basic" hemocompatibility on identical materials at different centers and attempt to rationalize. Assays for platelet adhesion, activation, aggregability and activation of the coagulation system in addition to an ex vivo patency assay were performed at four centers across Europe, using protocols favored by each center for determining the blood-contacting performance of a biomaterial. Three polymers were chosen for their expected blood response spanning the range of undesirable to desirable: ethylenevinylacetate (EVA), polyvinylchloride (PVC) and PVC modified with polyethylene oxide (PEO). The assays were ranked in terms of their efficacy compared to cost and simplicity. A correlation between assays was calculated, indicating the ability of one test to correctly determine the blood response compared to another. Some assays were unable to distinguish between materials, but of the assays which could, the materials were ranked in the following order: EVA; PVC; PVC-PEO, EVA producing the most undesirable response. It is concluded that many commonly used assays for determining hemocompatibility are inappropriate, but there are simple and reliable test methods available which correlate well with the more sophisticated protocols. PMID- 15348563 TI - Crystal growth and structure analysis of twin-free monoclinic hydroxyapatite. AB - Single crystals of hydroxyapatite were grown by a flux method using Ca(3)(PO(4))(2) and Ca(OH)(2) under 100 MPa of Ar gas. The crystals obtained had stoichiometric composition of Ca(10)(PO(4))(6)(OH)(2) and some of them were twin free single crystals. From X-ray diffraction analyses, the space group was confirmed to be monoclinic P2(1)/b with cell parameters a=0.9419(3) nm, b=1.8848(6) nm, c=0.6884(2) nm, and gamma=119.98(2) degrees. The detailed crystal structure was determined with a reliability factor R(w)=0.033; the O atoms of OH were located just on the 2(1) axis while the H atoms of OH occupied the positions a little deviated from the 2(1) axis. The origin of this structure was ascribed to the formation of hydrogen bonds between the H atoms of the OH ions and the specific O atoms of the PO(4) ions. PMID- 15348564 TI - Thermal and mechanical properties of biodegradable hydrophilic-hydrophobic hydrogels based on dextran and poly (lactic acid). AB - The thermal and mechanical properties of a new family of biodegradable hydrogels made of photocrosslinked dextran derivative of allyl isocyanate (dex-AI) and poly (D,L) lactide diacrylate macromer (PDLLAM) were studied. The changes of thermal and mechanical properties of the dex-AI/PDLLAM hydrogels as functions of dex-AI to PDLLAM composition ratio and immersion time in phosphate buffer solution at 37 degrees C were also investigated. Thermal property data showed that the chemical modification, crosslinking, swelling and hydrolytic degradation affected the glass transition and melting temperatures. Based on thermal data, no phase separation was observed in the bicomponent dex-AI/PDLLAM hydrogels. Mechanical property data showed that, by changing the composition ratio, dex-AI/PDLLAM hydrogels having a wide range of dry and swollen compression moduli could be obtained. The moduli of the dex-AI/DPLLAM hydrogels in dry state decreased with an increase in the PDLLAM composition due to the reduction in glass transition temperature of the hydrogels. The loss of mechanical strength in buffer solutions was attributed to the swelling-induced formation of 3D porous network structure in the early stage of immersion and the hydrolytic degradation of the PDLLAM in the late stage via the chain scission of ester linkages located in the PDLLAM backbone. Because swelling and degradation were composition dependent, the magnitude of the loss of mechanical strength was also composition-dependent. PMID- 15348565 TI - In situ polymerization into porous ceramics: a novel route to tough biomimetic materials. AB - A hydroxyapatite-based biomimetic composite, which is henceforth referred to as a synthetic bony material with high toughness characteristics, was prepared. It was obtained from a hydroxyapatite (HAp) skeleton with a relative porosity fraction of approximately 32 vol %, prepared by cold-isostatic-press compaction, followed by a sintering process, leading to a hydroxyapatite structure containing percolated submicrometer porosity channels. The percolated pores were infiltrated with a liquid mixture of epsilon-caprolactam monomer and an initiator, before homogeneous in situ polymerization to 6-nylon within the fully percolated pore structure was induced thermally. The final composite consisted of a dense interpenetrated hydroxyapatite/6-nylon network in a fraction approximately 68/30 vol %. The work of fracture value of the hybrid composite was found to be comparable with those found in two natural materials (bovine femur and nacre), which were also investigated under the same testing conditions. PMID- 15348566 TI - Effect of mixing ratio on bactericidal action of MgO-CaO powders. AB - MgO-CaO powders were prepared with different molar ratios (MgO/CaO) at 1400 degrees C for 2 h in air. By using the powder samples obtained, the change in bactericidal effect as a function of MgO-CaO composition was studied by colony count method. From the XRD measurements, it was found that CaO solid solution was formed by the replacement of Mg(2+) ion with larger Ca(2+) ion. However, no formation of MgO solid solution was observed. The average particle size and the specific surface area of the samples used in this study were about 0.2 microm and 10.5 m(2)g(-1), respectively. The pH values of physiological saline containing powder samples increased with the increase of CaO content, and the value reached 12.1 in sample with the molar ratio (MgO/CaO) of 0.25. From the results of bactericidal tests for Staphylococcus aureus, it was found that the bactericidal effect increased with the increase of CaO content in the samples. PMID- 15348567 TI - An experimental study of the dissemination of Titanium and Zirconium in the body. AB - Metallic implants can generate and release titanium oxide (TiO(2)) and zirconium oxide (ZrO(2)) to the tissues. These products can accumulate locally or disseminate systemically. The aim of the present study was to assess the distribution of TiO(2) and ZrO(2) administered intraperitoneally to rats. We used male Wistar rats of approximately 100 g body weight throughout the study. An intraperitoneal injection of a suspension of TiO(2) or ZrO(2) (16, 1600 and 16 x 10(3) mg/kg body weight) was administered. The animals were killed at 5-10 months post-administration by ether overdose. Samples of peritoneum, liver, kidney, lung and spleen were taken, fixed in formalin and routine processed for embedding in paraffin. One set of sections was stained with hematoxylin and eosin and another set was prepared unstained. The presence of titanium in the tissues was detected by X-ray diffraction crystallography. The histological analysis revealed the presence of abundant intracellular aggregates of metallic particles of Ti and Zr in peritoneum, liver, lung and spleen. The crystallographic study revealed the presence of anatasa. The dissemination of metallic particles from orthopedic or odontological implants would not be restricted to a local phenomenon. The particles also target vital organs. The distribution of these deposits over lengthy periods deserves meticulous attention given the clinical relevance of this phenomenon. PMID- 15348568 TI - In vitro response of primary rat osteoblasts to titania/hydroxyapatite coatings compared with transformed human osteoblast-like cells. AB - The biocompatibility of titania/hydroxyapatite (TiO(2)HA) composite coatings, at different ratio obtained by sol-gel process, was investigated studying the behavior of primary cultures of rat osteoblastic cells, isolated by femoral trabecular bone tissue. Moreover, the results have been compared with the response of human osteoblast-like MG63 cell line. Cytotoxicity of coatings was assessed by lactate dehydrogenase activity (LDH). The cellular behavior was analyzed by the cell proliferation (MTT test), cell morphology (SEM) and the biochemical markers evaluation of osteoblastic phenotype, such as alkaline phosphatase activity (ALP) and osteocalcin production. The results showed that TiO(2)/HA coatings have no toxic effects and seemed to be a good support for cell adhesion and proliferation. Moreover, these materials allowed the differentiation of osteoblasts, stimulating the expression of alkaline phosphatase activity. The responses of the primary rat osteoblasts and human osteoblast-like MG63 cell line grown onto these coatings were similar in terms of proliferation and ALP activity. Differences were found considering the osteocalcin production. The results show that these coatings, thanks to their chemical composition and the deposition technique, are very promising for the potential orthopedic and dental applications. PMID- 15348569 TI - Human biological reactions at the interface between bone tissue and polymethylmethacrylate cement. AB - This study investigated cytotoxicity of cement fragments harvested from two prosthesis revisions by the MTT test using L929 fibroblasts and human osteoblasts. The results did not show any toxicity of the extracts prepared after 48 and 78 months implantation. We consider that no MMA monomer has been released from the cement fragments. Histological studies on undecalcified samples harvested around revising prosthesis from 11 patients were used to evaluate tissue reactions at the bone-cement interface after 2-168 months implantation. Cement and prosthesis particles (5-35 microm) either dispersed or forming a layer were observed. A fibrous tissue layer, osteolysis, and osteonecrosis areas were observed at the interface. Besides, fibroblasts, macrophages, and multinucleated giant cells were also observed. New bone formation with osteoid, osteoblasts, and endochondral ossification with fibrocartilaginous tissue has been observed. The tissue reactions seemed to decrease with time. However, osseous trabeculae fractures were observed in the samples after 19 months. Although we consider that monomer toxicity, exothermic reaction, and particles formation may cause short term prosthesis loosening; the trabeculae fractures may be due to prosthesis and bone cement micromovements. This fractures and particles formation may cause long term prosthesis loosening. PMID- 15348570 TI - Microstructured microspheres of hydroxyapatite bioceramic. AB - Hydroxyapatite (HAP) particles having spherical geometry and 125-1000 microm in size range were prepared using a solid-in-water-in-oil (S/W/O) emulsion, cross linking technique. An aqueous solution of chitosan containing different loading of HAP was dispersed as droplet in liquid paraffin using a stabilizing agent. Cross-linking of chitosan was induced by adding appropriate amount of glutaraldehyde saturated toluene. Chitosan microspheres containing HAP were sintered at 1150 degrees C to obtain pure HAP microspheres. The spheres thus produced were examined by scanning electron microscopy. The percentage yield and size distributions of the spheres were also determined. PMID- 15348571 TI - Interaction between hydroxyapatite and proteins by liquid chromatography using simulated body fluids as eluents. AB - Hydroxyapatite (HAp) column chromatography for proteins using simulated body fluids (SBFs) as eluents was investigated in order to mimic the flow environment of human body fluid on the HAp surface in vivo. Acidic plasma proteins such as albumin, gamma-globulin, and fibrinogen having carboxyl groups with a negative charge were suggested to adsorb at positively charged sites (Ca sites) on the HAp surface. These acidic plasma proteins were not even eluted at higher inorganic ion molarities of human body fluid. Whereas basic proteins such as papin, cytochrome c, and lysozyme were easily eluted at lower inorganic ion molarities of human body fluid due to existing Na(+), Ca(2 +), Mg(2 +) and (CH(2)OH)(3)CNH(4)(+) ions in the eluents. The elution behavior of proteins using SBF in the presence or absence of glucose did not change. From these considerations, bone matrix proteins as acidic and neutral proteins would be considered an advantageous condition in order to adsorb on HAp in vivo. PMID- 15348572 TI - Rheological behavior of aqueous suspensions of hydroxyapatite (HAP). AB - The aim of this study is to characterize the influence of organic dispersants on the stability of hydroxyapatite (HAP) dispersions, using viscosity measurements, electrokinetic sonic amplitude (ESA) measurements and adsorption isotherms. Colloidal stability of aqueous suspensions of HAP as a function of solids loading and of the type and concentration of deflocculant has been studied. The interaction between particles and dispersant molecules was characterized. Adsorption measurements of the amount of dispersants on the particles surface, viscosity and electrophoretic measurements as a function of the dispersant concentration are correlated. Suspension parameters have been adjusted in order to obtain a high solids loading slurry with a plastic viscosity which will allow its use in colloidal processing. PMID- 15348573 TI - Fluoride release from model glass ionomer cements. AB - Glass ionomer cements (GICs) are an important class of biomedical material used extensively for color matched mercury free, dental restorations. GICs can release clinically beneficial amounts of fluoride and have acceptable handling properties which make them suitable as dental restoratives. The fluoride release of model GICs produced from specially synthesized fluoro-alumino-silicate glasses was studied. Nine glasses of varying fluoride content based on 4.5SiO(2)-3Al(2)O(3) 1.5P(2)O(5)-(5-Z)CaO-ZCaF(2) were synthesized and cement disks were prepared from them. The glass transition temperature reduced with increasing fluorine content of the glass. Fluoride ion release was measured into distilled water as a function of time for up to 140 days using a fluoride ion selective electrode. The quantity of fluoride released was found to be proportional to the fluorine content of the glass at all intervals time. The cumulative fluoride release was proportional to square root time. Substituting strontium for calcium in the glass had little influence on the fluoride release behavior of the cements. PMID- 15348574 TI - Enhancement of bonding strength by graded structure at interface between apatite layer and bioactive tantalum metal. AB - Tantalum metal is a candidate for use as an implant material in high load-bearing bony defects, due to its attractive features such as high fracture toughness and high workability. This metal, however, does not have bone-bonding ability, i.e. bioactivity, and therefore the development of bioactive tantalum metal is highly desirable. It is known that the essential prerequisite for an artificial material to show bioactivity is to form a bonelike apatite layer on its surface in the body environment. The same type of apatite layer is formed in a simulated body fluid (SBF) with inorganic ion concentrations nearly equal to those of human blood plasma. The present authors previously showed that the apatite formation on tantalum metal in SBF was remarkably accelerated by treatment with 0.5 M-NaOH aqueous solution and subsequent firing at 300 degrees C, while untreated tantalum metal spontaneously formed the same apatite after a long soaking period. In the present study, the bonding strength of the apatite layer to the substrate was quantitatively evaluated in comparison with that to the untreated tantalum metal. Adhesive strength was measured as an estimation of bonding strength, and the surface microstructure of both the substrates was characterized in order to discuss the difference in the bonding strength in terms of surface structure. The apatite layer formed on the NaOH- and heat-treated tantalum metal shows higher adhesive strength than that formed on the untreated metal. The amorphous sodium tantalate layer formed on the tantalum metal by NaOH and heat treatments, has a smooth graded structure where its concentration gradually changes from the surface into the interior metal. Smooth graded structure with complex of apatite is constructed after soaking in SBF. The higher bonding strength of the apatite layer formed on the treated metal is attributed to its smooth graded structure. PMID- 15348575 TI - Effect of hydrolysis on the phase evolution of water-based sol-gel hydroxyapatite and its application to bioactive coatings. AB - In a previous report, we demonstrated a successful synthesis of crystalline hydroxyapatite (HA) through the use of a water-based sol-gel process. It was shown that the apatite can be obtained at temperatures generally below 400 degrees C, providing a great advantage for practical bioactive coating purposes. The influence of hydrolysis of phosphorus sol solution on the phase evolution of the resulting HA is the focus of this investigation. Experimental results show that, in the absence of acid catalyst, a long-term hydrolysis, i.e. >4 h, is required for better evolution of apatitic phase. Such a phase evolution is mainly attributed to an increased concentration of apatitic phase, rather than improved crystallinity in the calcined gels. With the aid of acid catalyst, we found that a well-crystalline HA can be synthesized over a time period shorter by 2-3 orders of magnitude than those without catalyst, i.e. a few minutes. In almost all cases, a small amount of tricalcium phosphate (TCP) was detected, which may be explainable by the formation of oligomeric derivatives of the phosphorus sol during synthesis, where calcium phosphate derivatives with lower Ca/P ratio than stoichiometry can be developed. By selecting an optimal sol as a dipping source, highly-porous dental root specimens were coated and a thin, dense, adhesive (upon finger-nail scratching test) coating was achieved after calcinations at 375 degrees C. An in vitro test also shows a bioactive character of the coating. PMID- 15348576 TI - Biodegradation of hydrophilic-hydrophobic hydrogels and its effect on albumin release. AB - The objective of this study was to examine the effects of composition ratio of a new class of bicomponent biodegradable hydrogels and the molecular weights of the constituents on the hydrolytic degradability of the hydrogels and their release of bovine serum albumin (BSA). Biodegradable hydrogels were prepared from dextran derivative of allyl isocyanate (dex-AI) and poly (D,L) lactide diacrylate macromer (PDLLAM) over a wide range of dex-AI to PDLLAM composition ratio. The results obtained indicated that the hydrolytic degradation of these biodegradable hydrogels could be controlled by adjusting the composition ratio of dex-AI to PDLLAM or by changing their molecular weights. Along with the hydrogel degradation, water content of the hydrogels changed, and 3D porous network structure was observed. Generally, as the PDLLAM composition in the hydrogels increased, the rate of weight loss increased due to the hydrolytic degradation of the PDLLAM. The increase in molecular weights of either dex-AI or PDLLAM would decrease the degradation rate of the dex-AI/PDLLAM hydrogels. BSA release data correlated well with the hydrogel degradation profiles, suggesting that the extent and rate of BSA release would be mainly controled by hydrogel degradation. As the PDLLAM composition in the hydrogel increased, the extent and rate of BSA release also increased. An increase in the molecular weights of the hydrogel constituents, however, led to a decrease in BSA release. PMID- 15348577 TI - A novel urethane containing copolymer as a surface modification additive for blood contact materials. AB - Surface modification to develop a biomolecules-presenting surface is of interest, both from a scientific and an industrial point of view. In this research, a penta block-coupling polymer of warfarin-PEO-MDI-PEO-warfarin was specially designed as the surface modifying additive (SMA). The warfarin-modified polyurethane surfaces were then prepared by dip-coating method. Attenuated total reflection fourier transform infra-red (ATR-FTIR) spectra revealed that the urethane segments in the SMA could penetrate into the hard block of segmented polyurethane (SPU) via intermolecular hydrogen bonds. The X-ray photoelectron spectroscopy (XPS) results indicated that the intermolecular hydrogen bonds were strong enough to form stable warfarin-PEO composite surfaces in an aqueous environment. Fibrinogen and albumin adsorption onto unmodified and SMA-modified SPU was investigated by the (125)I-labeled method. The surface for attracting and reversibly binding albumin, which was proved to suppress the platelet adhesion and prolong the clotting time, has been developed by the simple coating of the novel SMA in SPU. PMID- 15348578 TI - Preparation and characterization of magnesium/carbonate co-substituted hydroxyapatites. AB - A new synthesis/processing method has been devised to produce magnesium/carbonate co-substituted hydroxyapatite ceramics that do not decompose to tricalcium phosphate (TCP) on sintering. Using this method, a series of magnesium/carbonate co-substituted hydroxyapatite (Mg/CO(3)-HA) compositions, containing between 0 and 0.35 wt % Mg and approximately 0.9 wt % CO(3) were prepared. Sintering the Mg/CO(3)-HA compositions in a CO(2)/H(2)O atmosphere yields a single crystalline phase that appears to be identical to stoichiometric HA. In contrast, when the compositions were prepared in the absence of carbonate and were sintered in air, the phase composition was a biphasic mixture of HA and TCP e.g. for 0.25 wt % Mg substitution the phase composition was approximately 60%HA/40% TCP. Clearly, both the synthesis route and the processing (i.e. sintering) route are of importance in the production of a single-phase Mg/CO(3)-HA ceramic. Fourier transform infrared (FTIR) spectroscopy has indicated that the Mg/CO(3)-HA ceramics still contained carbonate groups after sintering at 1200 degrees C. Chemical analysis by X-ray fluorescence spectroscopy (XRF) and C-H-N analysis has shown that the cation/anion molar ratio (i.e. [Ca+Mg]/[P+C/2]) of the different compositions were 1.68(+/-0.01), which is equivalent to the Ca/P molar ratio of stoichiometric HA. Although the magnesium/carbonate co-substitution had a positive effect in preventing phase decomposition during sintering, it appeared to have a negative effect on the densification of the MgCO(3)-HA ceramics, compared to stoichiometric HA. PMID- 15348579 TI - Structure and properties of Titanium-25 Niobium-x iron alloys. AB - The present work studies the effect of iron on microstructure, mechanical properties and corrosion behavior of Ti-25Nb based system with emphasis on improving strength/modulus ratio. Experimental data shows that cast Ti-25Nb-3Fe has a beta phase with a entirely of dendrite morphology. The bending strength/modulus ratio is 24.6 higher than Ti-6Al-4V (17.4) by 41.4% and than c.p. Ti (9.3) by 165%. The critical anodic current density of the metal in 37 degrees C Hank's solutions is lower than approximately 100 microA/cm(2). Ti-25Nb 3Fe alloy has a great potential for use as an implant material. PMID- 15348580 TI - Tests of biocompatibility of alpha-tocopherol with respect to the use as a stabilizer in ultrahigh molecular weight polyethylene for articulating surfaces in joint endoprostheses. AB - To inhibit the oxidation in vivo of hip-cups made of ultrahigh molecular weight polyethylene (UHMW-PE), the natural antioxidant alpha-tocopherol was added to the polymer. The added alpha-tocopherol may however undergo chemical transformations during manufacturing and sterilization by gamma-irradiation of hip-cups which may differ from human metabolism. Therefore, the question of the biocompatibility of the respective transformation products was investigated on test samples, which were prepared under the same conditions as applied for the production and sterilization of hip-cups. Thin plates (25 x 18 x 2 mm(3)) were fabricated out of test samples to investigate the cytotoxic activity according to EN 30993-5. In cytotoxicity testing, proliferation, mitochondrial activity and membrane integrity were not influenced by the material. In contrast, cell adhesion and cell spreading were diminished as shown with hemalum staining. In order to investigate the genotoxicity, the alpha-tocopherol and its transformation products were extracted from test specimens by n-heptane at 185 degrees C under nitrogen atmosphere. Then the n-heptane was evaporated in vacuo and the remaining alpha-tocopherol and its transformation products were dissolved in DMSO. The genotoxicity of this extract was then tested by the Ames-test according to DIN UA 12 (1995), which showed no indication for genotoxic activity. PMID- 15348581 TI - Effect of metal-oxide addition on the sintering of beta-calcium orthophosphate. AB - The effect of metal-oxide addition (1 approximately 10 mol %) on the sintering of beta-calcium orthophosphate (beta-Ca(3)(PO(4))(2)) was examined by pressureless sintering at 1070 degrees C for 5 h. The metal-oxide additives used were as follows: (i) monovalent metal-oxides, Li(2)O, Na(2)O, and K(2)O; (ii) divalent metal-oxides, MgO, CaO, SrO, and BaO; (iii) trivalent metal-oxides, Al(2)O(3) and Fe(2)O(3); and (iv) tetravalent metal-oxides, SiO(2), TiO(2), and ZrO(2). The relative densities of the sintered beta-Ca(3)(PO(4))(2) compacts were reduced with increasing amounts of monovalent and divalent metal-oxide additions, except for the case of MgO addition when the relative density remained with increasing amount of MgO up to 4 mol %. In the case of trivalent metal-oxide additions, the relative density of sintered beta-Ca(3)(PO(4))(2) compact was reduced with increasing amount of Al(2)O(3) addition, whereas it was enhanced with increasing amount of Fe(2)O(3) addition and reached 98.7% at 10 mol % Fe(2)O(3) addition. In the case of tetravalent metal-oxide additions, relative densities of the sintered beta-Ca(3)(PO(4))(2) compacts were slightly reduced with increasing amounts of SiO(2) and ZrO(2) additions, whereas no appreciable changes in the relative density were observed with increasing amount of TiO(2) addition. PMID- 15348582 TI - Thermal decomposition of synthesised carbonate hydroxyapatite. AB - Heat treatments are used when sintering hydroxyapatite to make porous blocks and granules and during plasma spraying of coatings. Calcium : phosphorus ratio is known to affect the thermal decomposition behavior of hydroxyapatite. Hydroxyapatite with carbonate ions substituted for phosphate ions is more similar in composition to bone mineral. While it has been shown that carbonate apatite may be sintered, relatively little is known about its high temperature stability. Various atmospheres have been used in investigations into the thermal stability of hydroxyapatites and carbonate hydroxyapatites, including nitrogen, wet carbon dioxide air, water vapor and wet oxygen, but few of these studies were directly comparable. Previous work has shown that loss of carbonate from CHA at high temperature is time dependent, which suggests that rapid high temperature treatment may prevent carbonate loss during processing. This study investigated the effect of dry carbon dioxide, carbon dioxide containing 3% water, nitrogen and nitrogen containing 3% water on the phase composition of hydroxyapatite containing between 1.0 and 11.5 wt % carbonate rapidly heated to temperatures of between 700 and 1400 degrees C. Carbonate ion substitution was observed to decrease the temperature at which crystallisation occurred to a minimum of 700 degrees C for 11.8 wt % carbonate apatite heated in wet atmospheres. Atmosphere was found to appreciably affect the crystallization temperature and phase transformations of carbonate apatite containing 7.8 wt % carbonate. In wet and dry carbon dioxide atmospheres, crystallisation began in this material at 1100 and 900 degrees C, betaTCP was formed at 1500 and 1300 degrees C respectively. The high temperature decomposition of carbonate hydroxyapatite would appear to depend on the composition of the apatite and the atmosphere in which it is heated. PMID- 15348583 TI - Comparative investigation of the surface properties of commercial titanium dental implants. Part I: chemical composition. AB - The surfaces of five commercially available titanium implants (Branemark Nobel Biocare, 3i ICE, 3i OSSEOTITE, ITI-TPS, and ITI-SLA) were compared by scanning electron microscopy, X-ray photoelectron spectroscopy, time-of-flight secondary ion mass spectroscopy. All five implant types were screw-shaped and fabricated from commercially pure (cp) titanium, but their surface properties differed both as regards surface morphology and surface chemical composition. The macro- and microstructure of the implant surfaces were investigated by scanning electron microscopy. The surfaces chemical composition was determined using the surface sensitive analytical techniques of X-ray photoelectron spectroscopy and time-of flight secondary ion spectrometry. Surface topographies were found to reflect the type of mechanical/chemical fabrication procedures applied by the manufacturers. The titanium oxide (passive) layer thickness was similar (5-6 nm) and typical for oxide films grown at or near room temperature. A variety of elements and chemical compounds not related to the metal composition were found on some implant types. They ranged from inorganic material such as sodium chloride to specific organic compounds believed to be due to contamination during fabrication or storage. The experimental findings are believed to make a contribution to a better understanding of the interplay between industrial fabrication procedure and physico-chemical implant surface properties. PMID- 15348584 TI - The effect of MgO on the solubility behavior and cell proliferation in a quaternary soluble phosphate based glass system. AB - This paper presents a systematic study of the MgO-CaO-Na(2)O-P(2)O(5) glass system, which has great potential to be used as temporary hard and soft tissue implant materials. An overall study of solubility behavior of ternary and quaternary-based phosphate glass system have been carried out in order to understand the out-leaching progress of different ions and to determine their effect on cell proliferation. Originally, soluble phosphate based glasses within the ternary glass system of Na(2)O-CaO-P(2)O(5) have been developed to create a simple baseline system. This paper, however, presents the development of this system by introducing magnesium oxide as a partial calcium oxide substitute and solubility behaviors as well as cell studies have been carried out to check the effect on magnesium ions. Glasses have been prepared via standard glass melting techniques and their solubility behavior has been tested in distilled water via simple weight loss, pH and ion measurements. The way the glasses dissolve is an inverse exponential behavior which is mirrored by the calcium ion release. Other ions show a less exponential behavior. The MTT test has been used to check preliminary in vitro studies on a human MG63 cell line and the result indicates that cell proliferation is increased for glasses with minimal CaO substitution. PMID- 15348585 TI - Interaction between bioactive glasses and human dentin. AB - This study explores the interaction between bioactive glasses and dentin from extracted human teeth in simulated oral conditions. Bioactive glasses in the Na(2)O-CaO-P(2)O(5)-SiO(2) and MgO-CaO-P(2)O(5)-SiO(2) systems were prepared as polished disks. Teeth were prepared by grinding to expose dentin and etching with phosphoric acid. A layer of saliva was placed between the two, and the pair was secured with an elastic band and immersed in saliva at 37 degrees C for 5, 21 or 42 days. The bioactive glasses adhered to dentin, while controls showed no such interaction. A continuous interface between the bioactive glass and dentin was imaged using cryogenic-scanning electron microscopy (SEM). However, after alcohol dehydration and critical point drying, fracture occurred due to stresses from dentin shrinkage. SEM investigations showed a microstructurally different material at the fractured interface. Chemical analyses revealed that ions from the glass penetrated into the dentin and that the surface of the glass in contact with the dentin was modified. Microdiffractometry showed the presence of apatite at the interface. Bonding appears to be due to an affinity of collagen for the glass surface and chemical interaction between the dentin and glass, leading to apatite formation at the interface. PMID- 15348586 TI - In vitro corrosion testing of PVD coatings applied to a surgical grade Co-Cr-Mo alloy. AB - Toxic effects and biological reaction of metallic corrosion and wear products are an important concern for metal on metal artificial joints. Corrosion tests were conducted to study the susceptibility to pitting and localized corrosion, with three coatings, CrN, TiN and DLC, applied to a wrought high carbon Co-Cr-Mo alloy substrate material. Corrosion testing involved the measurement of potential time transients during immersion in a physiological solution and cyclic polarization of specimen potentials into the transpassive range followed by reversal of the potential to scan in the cathodic direction to regain the rest potential E(rest). Resistance to pitting and localized corrosion was assessed by determining the transpassive breakdown potential E(bd) and if any hysteresis generated during the reverse cyclic scan may have caused crossover with the original anodic scan. Three different surface coating conditions were tested namely: (1) as-coated, (2) polished, and (3) indented to penetrate the coating by diamond pyramid hardness indentor. Results showed that all three coatings produced significant improvements in corrosion resistance compared to performance of the wrought cobalt alloy but that some corrosive attack to both the CrN and TiN coatings occurred and some risk of attack to the cobalt alloy substrate existed due to coating defects or when damage to the coating occurred. TiN coatings were highly effective in preventing corrosion provided they were thick enough to produce complete coverage. Thin TiN coatings displayed some tendency to encourage localized attack of the cobalt alloy at coating defects or where the coating suffered mechanical damage. CrN coatings underwent transpassive breakdown more easily and some degree of pitting at defects within the coating was observed, especially when the CrN coating was polished before the test. No corrosive attack of the cobalt alloy substrate was observed when the CrN coating was mechanically damaged by indentation. DLC coatings produced were much thinner than either of the other two coatings and proved to be rather fragile. They were less effective in preventing apparently high corrosion currents and possibly high rates of corrosion. PMID- 15348587 TI - A cultured living bone equivalent enhances bone formation when compared to a cell seeding approach. AB - The development of cell therapy methods to confer osteogenic potential to synthetic bone replacement materials has become common during the last years. At present, in the bone tissue engineering field, two different approaches use patient own cultured osteogenic cells in combination with a scaffold material to engineer autologous osteogenic grafts. One of the approaches consists of seeding cells on a suitable biomaterial, after which the construct is ready for implantation. In the other approach, the seeded cells are further cultured on the scaffold to obtain in vitro formed bone (extracellular matrix and cells), prior to implantation. In the present study, we investigated the in vivo osteogenic potential of both methods through the implantation of porous hydroxyapatite (HA) scaffolds coated with a layer of in vitro formed bone and porous HA scaffolds seeded with osteogenic cells. Results showed that as early as 2 days after implantation, de novo bone tissue was formed on scaffolds in which an in vitro bone-like tissue was cultured, while it was only detected on the cell seeded implants from 4 days onwards. In addition, after 4 days of implantation statistical analysis revealed a significantly higher amount of bone in the bone like tissue containing scaffolds as compared to cell seeded ones. PMID- 15348588 TI - Causes of titanium release from plate and screws implanted in rabbits. AB - To investigate the mechanism behind the release of metal from titanium implants in vivo, bone-plate-screw sets consisting of pure titanium were implanted into the legs of rabbits for 48 weeks. Four groups of experiments containing control were conducted: (1) The tibia cut artificially was fixed by one set of bone plate and screws, (2) the same set was implanted separately into muscles in the leg, (3) the set was fixed on the tibia and immediately retrieved, and (4) no implantation was performed. The amounts of titanium in all tissues from knee to ankle were quantified using atomic adsorption spectrometry. The ratio of amounts of titanium detected in the groups (1), (2), and (3) was 100 : 10 : 43. No titanium was detected in the group (4). Causes of the release of titanium in the group (1) include that in the groups (2) and (3). Major causes of titanium release were surgical handling in implantation and wear and/or fretting during experimental-term for 48 weeks. Titanium was also released in the absence of wear. No morphological abnormality was observed around tissue of the implant by biopsy at post-operation week 48. PMID- 15348589 TI - Plasma-sprayed zirconia bond coat as an intermediate layer for hydroxyapatite coating on titanium alloy substrate. AB - This study aims to strengthen the bonding at HA coating/Ti-6Al-4V interface by adding an intermediate ZrO(2) bond coat between them. The bonding strength of the HA/ZrO(2) coating was evaluated with the separately prepared HA coating as control. The phase, microstructure and chemistry, and surface roughness of the plasma-sprayed two-layer HA/ZrO(2) coating on Ti-6A1-4V substrate were investigated by X-ray diffractometry, scanning electron microscopy, and surfcorder, respectively. Experimental results indicate that the bonding strength increases from 28.6+/-3.22 MPa for HA coating to 36.2+/-3.02 MPa for HA/ZrO(2) composite coating. Elemental analysis employed on the surface of ZrO(2) bond coat, on which the HA top coat was first dissolved completely in HCl acid, reveals the sign of diffusion of calcium ions from HA to ZrO(2) bond coat. In addition, rougher surface morphology provided by ZrO(2) bond coat is also considered to aid in the bonding at HA/ZrO(2) interface. Similar coating system done by other researchers is compared and discussed. PMID- 15348590 TI - Self-assembly phenomenon of hydroxyapatite nanocrystals on chondroitin sulfate. AB - Self-assembly phenomenon of hydroxyapatite (HAp) nanocrystals on chondroitin sulfate (ChS) templates was investigated. A HAp/ChS hybrid was synthesized through a precipitation method with a calcium hydroxide suspension and phosphoric acid solution containing the ChS. The preferential alignment of the crystallographic c-axis of the HAp crystals parallel to the long axis direction of the ChS templates was observed to occur with the chemical interactions between the HAp crystals and the functional groups of the ChS. This phenomenon was interpreted by the crystallochemical specific nucleation and growth of the HAp crystals regulated by the pre-organized functional groups of the ChS template. Practical implication may involve a biomimetic artificial bone or cartilage can be made through a precipitation method with suitable bio-organics. PMID- 15348591 TI - Alloying of hydroxyapatite onto Ti6Al-4V by high power laser irradiation. AB - In the biomedical field, the synthetic hydroxyapatite [Ca(10)(PO)(4)(OH)(2)], with similarity to the inorganic component of bone but brittle, has been considered as the appropriate coating on stronger implant materials, such as metallic implants, for presenting a surface which is conductive to bone formation. Many industrial and laboratory techniques were developed to apply hydroxyapatite onto metallic substrates, such as electrophoretic deposition, ion sputtering, hot isostatic pressing, pulsed laser deposition and the only widely used method commercially available: plasma spraying. This work presents a new approach on how to bind calcium phosphate (CaP) to the Ti alloy with a well-known technique in the metallurgical field: laser surface alloying, in order to overcome the drawbacks of plasma spraying. The analysis of the results obtained and the description of the phenomena that take place in the coating process will complete this explorative study. PMID- 15348592 TI - Friction and wear of hydroxyapatite reinforced high density polyethylene against the stainless steel counterface. AB - Hydroxyapatite (HA) reinforced high density polyethylene (HDPE) was invented as a biomaterial for skeletal applications. In this investigation, tribological properties (e.g. wear rate and coefficient of friction) of unfilled HDPE and HA/HDPE composites were evaluated against the duplex stainless steel in dry and lubricated conditions, with distilled water or aqueous solutions of proteins (egg albumen or glucose) being lubricants. Wear tests were conducted in a custom-built test rig for HDPE and HA/HDPE containing up to 40 vol % of HA. It was found that HA/HDPE composites had lower coefficients of friction than unfilled HDPE under certain conditions. HA/HDPE also exhibited less severe fatigue failure marks than HDPE. The degradation and fatigue failure of HDPE due to the presence of proteins were severe for low speed wear testing (100 rpm) as compared to high speed wear testing (200 rpm). This was due possibly to the high shear rate at the contact which could remove any degraded film instantaneously at high sliding speed, while with a low sliding speed the build-up of a degraded layer of protein could occur. The degraded protein layer would stay at the contact for a longer time and mechanical activation would induce adverse reactions, weakening the surface layer of HDPE. Both egg albumen and glucose were found to be corrosive to steel and adversely reactive for HDPE and HA/HDPE composites. The wear modes observed were similar to that of ultra-high molecular weight polyethylene. Specimens tested with egg albumen also displayed higher wear rates, which was again attributed to corrosion accelerated wear. PMID- 15348593 TI - Strength and fatigue of polyacid-modified restorative materials (compomers). AB - The purpose of this study was to compare the fatigue behavior of a hybrid composite, four compomers, and two viscous glass ionomers after short- and long term soaking in distilled water. Bars with dimensions of 30 mm x 2 mm x 2 mm were formed in stainless steel molds, finished with 600 grit SiC, then soaked for either 24 h or one year, and tested in 3-point flexure at stressing rates between 0.001 and 2500 MPa s(-1). Data were plotted as fracture stress vs. stressing rate, and the exponent N in the power law for crack growth rate was computed from the slope of these plots. All compomer and resin composite materials tested exhibited subcritical crack growth (i.e. a reduction in strength with a decrease in stressing rate). Soaking lowered the slope for all materials, that is, increased the crack propagation rate. F2000 exhibited an increase in fracture strength while Hytac and Compoglass exhibited a decrease following long-term soaking in distilled water. Crack-growth exponents obtained from these fatigue data were used to estimate the stresses which would result in a five-year lifetime for these materials. PMID- 15348594 TI - Electrochemical impedance spectroscopy study of high-palladium dental alloys. Part I: behavior at open-circuit potential. AB - Electrochemical impedance spectroscopy (EIS) was used to study the in vitro corrosion of three representative high-palladium alloys and a gold-palladium alloy for comparison. The corrosion resistances (measured as the charge transfer resistance R(CT) from an equivalent circuit) of the high-palladium alloys and the gold-palladium alloy were comparable in simulated body fluid and oral environments, and under simulated dental plaque. The great similarity in corrosion behavior for the three high-palladium alloys is largely attributed to their substantial palladium content and passivity in the laboratory test media, and possibly to their similar structure at the submicron level. Differences in composition and microstructure at the micron level and greater, including the effects of heat treatment simulating the firing cycles for dental porcelain, do not have noteworthy effects on the in vitro corrosion of the three high-palladium alloys. Good accuracy and convenience of extracting corrosion characteristics from equivalent circuit modeling, along with the capability of providing intrinsic information about the corrosion mechanism, enable EIS to be an excellent alternative method to conventional potentiodynamic polarization for evaluating the corrosion behavior of noble dental alloys. PMID- 15348595 TI - Electrochemical impedance spectroscopy study of high-palladium dental alloys. Part II: behavior at active and passive potentials. AB - Electrochemical impedance spectroscopic (EIS) analyses were performed on three high-palladium alloys and a gold-palladium alloy at active and passive potentials in five electrolytes that simulated body fluid and oral environmental conditions. All four alloys were previously found to have excellent corrosion resistance in these in vitro environments. Before performing the EIS analyses, alloy specimens were subjected to a clinically relevant heat treatment that simulated the firing cycles for a dental porcelain. It was found that the EIS spectra varied with test potential and electrolyte. Diffusional effects, related to the dealloying and subsequent surface enrichment in palladium of the high-palladium alloys, along with species adsorption and passivation, were revealed at both active and passive potentials, although these effects were more evident at the passive potentials. PMID- 15348596 TI - The static and cyclic strength of a bone-cement bond. AB - Four-point bending static and fatigue tests were carried out on bone-cement bonds. The effects of the pressurization and the washing of the bone joint face on the bond strength were investigated. The results are summarized as follows. When the bond surface of cancellous bone is washed prior to the application of the bone cement, both the static and fatigue strengths of the bond are increased relative to the corresponding properties of unwashed bone-cement bonds. From observations of bone-cement interfaces as well as the fracture surfaces of bone cement specimens, it has been determined that bone cement was able to infiltrate into fine holes present in washed cancellous bone. However, such infiltration occurred to a much lesser degree in the case of unwashed cancellous bone. Increasing the molding pressure during the time of cement application to the bone from 39200 to 117600 Pa had a beneficial effect on the bending strength and fatigue properties, particularly in the case of washed bone cement specimens. An increase in molding pressure also resulted in a reduction in the amount of scatter in test results. PMID- 15348597 TI - Characterization of surface oxide films on titanium and bioactivity. AB - Biological properties of titanium implant depend on its surface oxide film. In the present study, the surface oxide films on titanium were characterized and the relationship between the characterization and bioactivity of titanium was studied. The surface oxide films on titanium were obtained by heat-treatment in different oxidation atmospheres, such as air, oxygen and water vapor. The bioactivity of heat-treated titanium plates was investigated by immersion test in a supersaturated calcium phosphate solution. The surface roughness, energy morphology, chemical composition and crystal structure were used to characterize the titanium surfaces. The characterization was performed using profilometer, scanning electronic microscopy, ssesile drop method, X-ray photoelectron spectroscopy, common Bragg X-ray diffraction and sample tilting X-ray diffraction. Percentage of surface hydroxyl groups was determined by X-ray photoelectron spectroscopy analysis for titanium plates and density of surface hydroxyl groups was measured by chemical method for titanium powders. The results indicated that heat-treatment uniformly roughened the titanium surface and increased surface energy. After heat-treatment the surface titanium oxide was predominantly rutile TiO(2), and crystal planes in the rutile films preferentially orientated in (1 1 0) plane with the highest density of titanium ions. Heat-treatment increased the amount of surface hydroxyl groups on titanium. The different oxidation atmospheres resulted in different percentages of oxygen species in TiO(2), in physisorbed water and acidic hydroxyl groups, and in basic hydroxyl groups on the titanium surfaces. The immersion test in the supersaturated calcium phosphate solution showed that apatite spontaneously formed on to the rutile films. This revealed that rutile could be bioactivated. The analyses for the apatite coatings confirmed that the surface characterization of titanium has strong effect on bioactivity of titanium. The bioactivity of the rutile films on titanium was related not only to their surface basic hydroxyl groups, but also to acidic hydroxyl groups, and surface energy. Heat-treatment endowed titanium with bioactivity by increasing the amount of surface hydroxyl groups on titanium and its surface energy. PMID- 15348598 TI - Surface oxidation of polyethylene fiber reinforced polyolefin biomedical composites and its effect on cell attachment. AB - Three different compositions of butene-ethylene copolymer composites reinforced by polyethylene fibers and produced by filament winding are potentially suitable for biomedical applications. This study examines the effect of various processing and finishing conditions and of sterilization on the extent and composition of surface oxidation. An XPS analysis revealed only insignificant differences between the various treatments, while fibroblast cell attachment tests indicated good attachment with no signs of cytotoxity or cell degeneration for any of the materials. PMID- 15348599 TI - Vibrational and thermal study on the in vitro and in vivo degradation of a poly(lactic acid)-based bioabsorbable periodontal membrane. AB - Fourier transform Raman (FT-Raman), attenuated total reflection/Fourier transform infrared (ATR/FT-IR) spectra and differential scanning calorimetry (DSC) measurements were performed on a poly(lactic acid)-based biodegradable periodontal membrane in order to study its in vitro and in vivo degradation mechanism and kinetics. For this purpose, the hydrolitic in vitro degradation of the membrane was investigated in two aqueous media: saline phosphate buffer (SPB, pH=7.4) and 0.01 M NaOH solution. Moreover, a membrane implanted in vivo for four weeks for treatment of contiguous vertical bony defects, was examined. Vibrational and thermal measurements show that the membrane has a prevalently amorphous structure and is composed of low molecular weight polymeric chains. The degradation is faster in NaOH solution than in SPB and occurs heterogeneously without any significative increase in crystallinity. The DSC and spectroscopic measurements are discussed in comparison with the trend of % weight loss and show a progressive decrease in molecular weight. Regarding the Raman analysis, the I(875)/I(1452) intensity ratio was identified as a marker of the degree of degradation. Regarding the in vivo degradation, the presence, spectroscopically revealed, of a biological component entrapped in the membrane proves the good integration of the membrane with the surrounding tissues. The membrane seems to degrade faster in vivo than in vitro. A comparison with the degradation mechanism and kinetics of a periodontal membrane previously studied, Vicryl periodontal mesh, is made. PMID- 15348600 TI - Mechanical effects of increases in the load applied in uniaxial and biaxial tensile testing. Part II. Porcine pericardium. AB - The mechanical behavior of porcine pericardium was analyzed to compare it with that of calf pericardium employed in valve leaflets for cardiac bioprostheses. Forty samples of pericardium were subjected to uniaxial tensile testing, 20 as controls and 20 exposed to loads increasing stepwise from 0.5 to 1.5 kg and to 3 kg, and thereafter to rupture, with a return to zero load between each new increment. Another 20 samples were used in biaxial tensile tests involving the application of loads increasing stepwise (to 0.5, 1.5, 3 and 5 kg) until rupture with a zero-load interval before each increment. The ultimate stresses were very similar, showing no statistically significant differences when compared in terms of type of assay, controls and study samples or region of pericardial tissue being tested. In the stepwise biaxial assays, the mean stresses at rupture were also very homogeneous. Using morphological and mechanical criteria for sample selection, it was possible to obtain mathematical fits for the stress/strain relationship, with excellent coefficients of determination. The relationship between the area under the stress/strain curve and the load applied or the strain observed was also studied in the biaxial assay as an equivalent to the cycles of hysteresis produced in the test. The increment in the area under the curve (the energy consumed) may be a good parameter for assessing the changes in the collagen fiber architecture of the pericardial tissue, changes that may help to detect early failure. PMID- 15348601 TI - Correlation between structure and compressive strength in a reticulated glass reinforced hydroxyapatite foam. AB - Glass-reinforced hydroxyapatite (HA) foams were produced using reticulated foam technology using a polyurethane template with two different pore size distributions. The mechanical properties were evaluated and the structure analyzed through density measurements, image analysis, X-ray diffraction (XRD) and scanning electron microscopy (SEM). For the mechanical properties, the use of a glass significantly improved the ultimate compressive strength (UCS) as did the use of a second coating. All the samples tested showed the classic three regions characteristic of an elastic brittle foam. From the density measurements, after application of a correction to compensate for the closed porosity, the bulk and apparent density showed a 1 : 1 correlation. When relative bulk density was plotted against UCS, a non-linear relationship was found characteristic of an isotropic open celled material. It was found by image analysis that the pore size distribution did not change and there was no degradation of the macrostructure when replicating the ceramic from the initial polyurethane template during processing. However, the pore size distributions did shift to a lower size by about 0.5 mm due to the firing process. The ceramic foams were found to exhibit mechanical properties typical of isotropic open cellular foams. PMID- 15348602 TI - Hydrothermal synthesis of hydroxyapatite from natural source. AB - Dicalcium phosphate dihydrate (DCPD) and anhydrous dicalcium phosphate (DCPA) extracted from a by-product in a manufacture of gelatin from bovine bone are used as starting materials for the hydrothermal synthesis of hydroxyapatite (HA) at temperatures of 160 and 200 degrees C under vapor pressures of 1 and 2 MPa, respectively. The suspension of DCPA with water gives a mixture of DCPA and HA as product but after adjusting the Ca/P molar ratio to 1.50 and 1.67 by addition of Ca(OH)(2), a single phase of HA with needle-like crystals is obtained. DCPA whiskers are produced in the suspension of DCPD with water but, on addition of Ca(OH)(2), the product obtained is again, a mixture of DCPA and HA. The coexistence of DCPA and HA is observed not only in the acidic region at pH 4.6 which is close to the known quasi-invariant point but also in the basic region at pH 12.3. Without addition of Ca(OH)(2) to the system, the complete conversion of DCPA or DCPD to HA is not possible. PMID- 15348603 TI - Electrochemical and microstructural studies of tantalum and its oxide films for biomedical applications in endovascular surgery. AB - The most popular coronary stents are made of 316L stainless steel and self expandable Nitinol. Nevertheless, Ta has already been used to make stents for endovascular surgery and may constitute a good alternative to the other materials because of its higher corrosion resistance and radio-opacity property, which may facilitate the follow-up of stent catheterization. The characterization of Ta and its natural passive oxide films has been performed in a 0.15 M NaCl solution (simulated body fluid - SBF) using anodic polarizations, electrochemical impedance spectroscopy and photoelectrochemical techniques. Changes in microstructure have been observed by atomic force microscopy (AFM). Polarization curves show the existence of a current density increase between 1.40 and 1.80 V. Bode complex plots show that some perturbation of the film occurred in this potential interval which may be associated with a decrease in polarization resistance, Rp, indicating that the film may be less resistant to corrosive attack. Mott-Schottky capacity measurements show that the density of donors, Nd, varies with polarization. The optical band gap, E(g), which is equal to 4.1 eV did not show variations in our experiments. The localized formation on the electrode surface, in the above potential interval of a Ta compound (possibly an oxide-hydroxide) was observed by AFM, and this may explain the appearance of the current density peak and capacity behavior at those potentials. PMID- 15348604 TI - Fretting wear study of surface modified Ni-Ti shape memory alloy. AB - A combination of shape memory characteristics, pseudoelasticity, and good damping properties make near-equiatomic nickel-titanium (Ni-Ti) alloy a desirable candidate material for certain biomedical device applications. The alloy has moderately good wear resistance, however, further improvements in this regard would be beneficial from the perspective of reducing wear debris generation, improving biocompatibility, and preventing failure during service. Fretting wear tests of Ni-Ti in both austenitic and martensitic microstructural conditions were performed with the goal of simulating wear which medical devices such as stents may experience during surgical implantation or service. The tests were performed using a stainless steel stylus counter-wearing surface under dry conditions and also with artificial plasma containing 80 g/L albumen protein as lubricant. Additionally, the research explores the feasibility of surface modification by sequential ion implantation with argon and oxygen to enhance the wear characteristics of the Ni-Ti alloy. Each of these implantations was performed to a dose of 3 x 10(17) atom/cm(2) and an energy of 50 kV, using the plasma source ion implantation process. Improvements in wear resistance were observed for the austenitic samples implanted with argon and oxygen. Ion implantation with argon also reduced the surface Ni content with respect to Ti due to differential sputtering rates of the two elements, an effect that points toward improved biocompatibility. PMID- 15348605 TI - Physical and mechanical properties evaluation of Acropora palmata coralline species for bone substitution applications. AB - The search for ideal materials for bone substitution has been a challenge for many decades. Numerous natural and synthetic materials have been studied. For this application, exoskeletons of coral have been considered a good alternative given its tendency to resorption, biocompatibility and similarity to the mineral bone phase. Very few studies of these materials consider a detailed analysis of the structure-property relationship. The purpose of this work was to carry out the microstructural characterization of a coralline species named Acropora palmata and the determination of the mechanical and physico-chemical properties. Measurements of hardness, compressive strength, bulk density and apparent porosity were performed. From these results it was determined that this marine coral species could be an alternative xenograft due to its mechanical properties and osteoconductive nature. PMID- 15348606 TI - Effect of vinyl acetate content on the sintering behavior of hydroxyapatite ethylene vinyl acetate copolymer composites. AB - Ethylene vinyl acetate copolymer (EVA) alone could be used as a binder material for the fabrication of hydroxyapatite (HAP) into intricate shapes for various bone substitute applications. It was observed that as the vinyl acetate content in the polymer was increased from 12 to 28 wt % an increase in the sintered density of the HAP was observed. Retention of the shapes of HAP in the molded form was also observed. PMID- 15348607 TI - Gelatin nanoparticles produced by a simple W/O emulsion as delivery system for methotrexate. AB - Biodegradable hydrophilic gelatin nanoparticles, containing different initial amounts of methotrexate (MTX), were prepared using a simple solvent evaporation technique based on a single water-in-oil emulsion and stabilized by the use of glutaraldehyde as cross-linking agent. The effects of several parameters on particle size, drug encapsulation efficiency and drug release were investigated. Size and shape of the nanoparticles were examined by scanning electron microscopy. The release of MTX was monitored in vitro and the mechanism of release was studied. Particles with a mean diameter of 100-200 nm were produced, which were able to release MTX following a diffusion-controlled mechanism of release. It was observed that the initial amount of MTX used for sample loading did not have any effect on the pattern of release, while it affected the amount of drug entrapped into the nanoparticles and also both the release rate and the total amount of drug released. PMID- 15348608 TI - The cell attachment and morphology of neonatal rat calvarial osteoblasts on the surface of Ti-6Al-4V and plasma-sprayed HA coating: effect of surface roughness and serum contents. AB - The biocompatibility of material plays an important role in the bone-implant interface for the prosthetic implant fixation. The biocompatibility of implants is associated with the chemical composition, surface topography, surface energy and surface roughness of biomaterials. The effects of two factors, surface roughness and serum contents, on osteoblast behavior at the surface of Ti-6Al-4V and plasma sprayed HA coating were investigated in the experiment. The osteoblasts derived from neonatal rat calvarial were cultured in Dulbecco's modified Eagle medium (DMEM) with fetal bovine serum (FBS) on the surface of polished Ti-6Al-4V (Ti-p), grit-blasted Ti-6Al-4V (Ti-b), polished HA coating (HAC-p), and as-sprayed HA coating (HAC). Under culture medium containing 4% FBS, the level of cell attachment to the polished surface is significantly higher than the rough surface of the same experimental materials during all culture periods. Increasing the contents of FBS up to 10%, the difference of osteoblast attachment is not found between Ti-p and Ti-b. Under 4% serum condition, the cell morphology attached to smooth surfaces (Ti-p and HAC-p) is spread faster and are more flattened than the one to rough surface of the same experimental materials by SEM. After 24 h culture, the corroded cracks are easily observed at the surface of polished HA coatings, and the cell morphology on HAC-p coatings are elongated and less flattened compared with Ti-p. The result is consistent with statistical difference of cell attachment between Ti-p and HAC-p under 4% serum condition. PMID- 15348609 TI - Studies of polyurethane urea bands for ACL reconstruction. AB - The present report describes the mechanical tests, in vitro and in vivo studies of a poly(urethane urea) (PUUR) intended for clinical use in anterior cruciate ligament (ACL) reconstruction. In the mechanical tests, no evidence of severe fatigue was observed after repeated cyclic loading. Testings for mutagenicity and delayed contact hypersensitivity were found negative. Three in vivo studies were performed in rabbits and minipigs. Altogether 35 rabbits were operated upon in (1) an intraarticular implantation study, performed to evaluate the soft tissue response to woven bands and fiber bundles of PUUR and (2) a rabbit ACL study, examining the function of the PUUR ACL replacement and the tissue response to the material. In a third study, PUUR ACL replacement in minipigs was evaluated. Taken together, ingrowth of connective tissue in close contact with the PUUR fibers was detected both in rabbits and minipigs. The first clear histological signs of degradation of the polymer was detected after 24 months. In conclusion, the evaluated mechanical properties of the PUUR band correspond to those of the mature, human ACL. Furthermore, both from a histological and functional point of view, the PUUR woven band show interesting properties for future clinical ACL reconstructions. PMID- 15348610 TI - Fatigue studies of high-palladium dental casting alloys: Part I. Fatigue limits and fracture characteristics. AB - The fatigue limits and fracture characteristics for a Pd-Cu-Ga alloy and a Pd-Ga alloy were studied. The alloys were cast into tensile test bars with gauge diameter of 3 mm and gauge length of 15 mm, and the surfaces of the castings were neither air-abraded nor polished after removal from the investment. Specimens were prepared from all-new metal (not previously melted), a combination of 50% new metal and 50% old metal (previously melted one time) and 100% old metal. The cast bars were subjected to heat treatment simulating the complete firing cycles for dental porcelain, and fatigued in air at room temperature under uniaxial tension-compression stress at 10 Hz and a ratio of tensile stress amplitude to compressive stress amplitude (R-ratio) of -1. The alloy microstructures and fracture surfaces were examined with a scanning electron microscope (SEM). Results showed that the fatigue limits at 2 x 10(6)cycles of the Pd-Cu-Ga and Pd Ga alloys were approximately 0.20 and 0.15 of their 0.1% yield strength (YS) in tension, respectively. The fatigue resistance for specimens from both alloys containing 50% old metal and 50% new metal was comparable to that of specimens containing all-new metal, although this decreased dramatically for Pd-Cu-Ga alloy specimens containing all-old metal. The fatigue resistance of the Pd-Cu-Ga alloy subjected to heat treatment simulating the porcelain firing cycles was not adversely affected by remnants of the original as-cast dendritic microstructure that remained in the relatively large test specimens. A longer heat treatment than recommended by the manufacturer for the porcelain firing cycles is needed to completely eliminate the as-cast dendritic structure in these specimens. The Pd Cu-Ga alloy exhibited superior fatigue resistance to the Pd-Ga alloy, which has an equiaxed-grain microstructure and lower yield strength. PMID- 15348611 TI - Fatigue studies of high-palladium dental casting alloys: Part II. Transmission electron microscopic observations. AB - The microstructures of two representative high-palladium dental alloys, a Pd-Cu Ga alloy and a Pd-Ga alloy, which had been subjected to cyclic fatigue in uniaxial tension were investigated by transmission electron microscopy (TEM). Two different mechanisms were found to dominate microplastic deformation during fatigue: twinning in the Pd-Cu-Ga alloy, and planar slip of dislocations in the Pd-Ga alloy. In addition, stress-induced precipitation occurred in the Pd-Ga alloy during cyclic loading. Heat treatment simulating the firing cycles for dental porcelain resulted in the formation of a previously unreported bcc phase in the Pd-Cu-Ga alloy, and in the elimination of the characteristic tweed structure found in the Pd-Ga alloy for the as-cast condition. PMID- 15348612 TI - In vivo response of strontium and zinc-based ionomeric cement implants in bone. AB - In this study the osteoconductive properties of strontium based ionomeric cements (ICs) named, LG125 and LG119, as well as zinc-based ICs, designated by LG130 and LG132, were compared. Wet ICs were surgically implanted into the femora of weaned Wistar rats for 4 weeks. To assess the percentage osseointegration the perimeter of the implant and the perimeter of bone in contact with the implant were measured using a pointer (the length of bone/implant interfacial contact). Osteoconduction was determined by taking six points at random around the perimeter of each ionomeric rod measuring the thickness of newly formed bone. The degree of osteoconduction was taken as the average thickness of new bone produced on the implant surface. It was found that osteoconduction was greatest in the strontium based IC implant LG125. From these studies it can be concluded that the composition LG125 might provide a useful purpose as a bone cement. PMID- 15348613 TI - Mechanical effects of increases in the load applied in uniaxial and biaxial tensile testing: Part I. Calf pericardium. AB - The authors analyzed the mechanical behavior of the calf pericardium employed in the construction of valve leaflets for cardiac bioprostheses. Forty samples of pericardium were subjected to uniaxial tensile testing, 20 as controls and 20 exposed to loads increasing stepwise until rupture, with a return to zero load between each new increment. Another 20 samples were used similarly in biaxial tensile tests involving loads increasing stepwise until rupture, again returning to zero load between steps. The ultimate stresses in the uniaxial study were very similar and were not influenced by the region of pericardial tissue being tested or the increments in load to which the tissue was exposed. The mean stresses at rupture in the stepwise biaxial assays were significantly greater (p<0.01). Using morphological and mechanical criteria for sample selection, it was possible to obtain mathematical fits for the stress/strain relationship in both types of assays, with excellent coefficients of determination (R (2)>0.90). In uniaxial tests in which the selection criteria were not applied, the correlation improved as the load increased, a phenomenon that did not occur in the biaxial studies. The values varied throughout the different cycles, adopting exponential forms when the strain was greatest. These variations, which demonstrate that the increase in the energy consumed is a function of the stress applied and of the strain produced, should be good parameters for assessing the changes in the collagen fiber architecture of pericardial tissue subjected to cyclic stress, and may help to detect early failure. PMID- 15348614 TI - Strength retention of drawn self-reinforced polyglycolide rods and fixation properties of the distal femoral osteotomies with these rods. An experimental study on rats. AB - Drawn self-reinforced polyglycolide (SR-PGA) rods, O 2 mm and 26 mm long, were implanted in the dorsal subcutaneus tissue of 16 rats. Osteotomies of the distal femur were fixed with SR-PGA rods (2 mm by 15 mm) in another 38 rats. The follow up times varied from one week to one year. After sacrifice, three-point bending and shear tests were performed for subcutaneously placed rods. Radiological, histological, histomorphometrical, microradiographic, and oxytetracycline fluorescence studies of osteotomized and intact control femora were also performed. At three weeks the flexural strength of the rods was 50% of the initial value, and the flexural modulus was 46% of the initial value. Five osteotomy specimens had to be excluded due to dislocation or non-union. One of the 33 evaluated osteotomy specimens showed signs of postoperative infection. Thirty-two osteotomies healed uneventfully. No gross signs of inflammatory or foreign-body reaction were observed. The amount of osteoid surface and active osteoid formation surface reached their highest value in the histomorphometrical analysis at 24 weeks. The present investigation demonstrated that the mechanical strength and fixation properties of the drawn SR-PGA rods are suitable for fixation of cancellous bone osteotomies in rats. The present article is the first report on the successful application of drawn SR-PGA rods for fixation of cancellous bone osteotomies. PMID- 15348615 TI - Processing and mechanical properties of autogenous titanium implant materials. AB - Pure titanium and some of its alloys are currently considered as the most attractive metallic materials for biomedical applications due to their excellent mechanical properties, corrosion resistance, and biocompatibility. It has been demonstrated that titanium and titanium alloys are well accepted by human tissues as compared to other metals such as SUS316L stainless steel and Co-Cr-Mo type alloy. In the present study, highly porous titanium foams with porosities 1 indicates the acute toxicity of the tested substance (necrosis). The extracts of chromium, cobalt and nickel had a cytotoxic effect on the mononuclear cells; high concentrations of cobalt and nickel produced cell necrosis, whereas by lowering the extract concentration apoptotic phenomena were observed. High chromium concentrations can induce cell death by apoptosis. Our data suggest that when large amounts of nickel and cobalt are released from implanted metal devices, necrosis is produced and consequently a strong inflammatory tissue reaction is likely to occur. The release of either chromium or limited amounts of nickel and cobalt induces toxicity characterized by apoptotic phenomena, which allows an adaptation of the tissue to the implant. PMID- 15348700 TI - Glutaraldehyde-fixed biological tissue calcification: effectiveness of mitigation by dimethylsulphoxide. AB - The conditions defining the extent of dimethylsulphoxide (DMSO) effectiveness in mitigating calcification of glutaraldehyde (GA)-fixed tissue have been evaluated. Exposure of GA-fixed tissue porcine aortic valve cusps to low concentrations of DMSO does not impart calcification inhibitory activity. Mitigation in calcification becomes evident only as the DMSO component nears 100% and is best when neat DMSO is used. In all instances, regardless of the DMSO concentration, exposure to DMSO resulted in an increase in the tissue shrinkage temperature, attributed to further cross-linking in the tissue. Histological examination of samples before implantation indicate some deleterious effects to the tissue, the degree dependent on concentration, time and temperature of DMSO exposure. The results of this study suggest that treatment of GA-fixed tissue with high concentrations of DMSO for a short duration at a lowered temperature could give a bioprosthesis that has good mitigating calcification properties with retention of tissue integrity. PMID- 15348701 TI - A specific quantitative assay for collagen synthesis by cells seeded in collagen based biomaterials using sirius red F3B precipitation. AB - The measurement of collagen synthesis by seeded cells in vitro is a prerequisite for the assessment of biocompatibility of many biomaterials. Existing methods are either complicated or not applicable to systems utilizing collagen-based materials, and the development of a rapid and simple technique would be an advantage. In the current paper, a method is described which relies on the radiolabelling of newly synthesized protein with [3H]-proline followed by specific precipitation of collagen using 1% sirius red dissolved in water. The results indicate that collagen binding to sirius red is unaffected by using water rather than picric acid as a solvent and the dye binds in a similar fashion to collagen type I, II and III. Cycloheximide treatment of the gels indicated that precipitated [3H]-proline was restricted to macromolecular protein. Collagenase treatment eradicated labelled precipitation formation when using 1% sirius red in water, indicating a high degree of specificity for collagen whilst specificity was poor when using 1% sirius red in picric acid. The method described is both simple and rapid and shows a high degree of specificity and sensitivity. For these reasons it is highly suited for the assessment of collagen synthesis by cells in collagen-based materials. PMID- 15348702 TI - Influence of temperature and additives on the microstructure and sintering behaviour of hydroxyapatites with different Ca/P ratios. AB - Sintering of two hydroxyapatite (HA) samples with different Ca/P ratios was studied as a function of thermal pretreatments, sintering temperature and additives (0-0.6 ion % Li+ or 0-5 ion % Mg2+). The samples were sintered in air and characterized by density measurements, scanning electron microscopy, differential thermal analysis, X-ray diffraction and dilatometry. Upon sintering, samples with Ca/P ratio of 1.51 (HA C) transformed to beta-Ca3(PO4)2 and Ca10(PO4)6(OH)2, resulting in materials with low densities and containing agglomerated beta-Ca3(PO4)2 when sintered above 1200 degrees C. Samples with a Ca/P ratio of 1.77 (HA S), without beta-Ca3(PO4)2, showed better sinterability and homogeneous microstructures. Li+ additions favoured liquied-phase sintering and reduced the beta-Ca3(PO4)2 content in sintered materials. Mg2+ additions did not result in higher densities, but inhibited the hydroxyapatite grain growth rate. A significant percentage of the added Mg2+ was incorporated into the beta Ca3(PO4)2 structure. PMID- 15348703 TI - Systemic titanium levels in rabbits with a titanium implant in the absence of wear. AB - All non-noble metals and alloys will release metallic species into the body. This raises the issue of amount and fate, i.e. transport and storage, of these metal dissolution products. For titanium, the nature and extent of these systemic effects remain mostly unknown. In this study we investigated titanium levels in alleged target tissues in rabbits, both with and without a titanium implant functioning in the absence of wear, and compare these results to the limited body of literature concerning systemic levels of titanium. Titanium fibre felts were implanted into the tibia of rabbits. At various time points, lung, spleen, and muscle samples were collected from these rabbits as well as two groups of control rabbits. The samples were analysed for titanium concentration using electrothermal atomic absorption spectrophotometry. The data for the implant groups show that titanium levels in these tissues do not increase in comparison with controls up to 1 y after implantation. PMID- 15348704 TI - An animal study on the bone behaviour of Ca-P-coated implants: influence of implant location. AB - Four different implant materials were installed into the mandibular corner of goats to investigate the trabecular bone response in a mainly unloaded model. The implants were installed using a standardized technique and were left in situ for 12 weeks. One goat had to be sacrificed after surgery because of a broken rib; the other animals healed uneventfully. After sacrifice of the animals, the bone response to the uncoated and the three different Ca-P implants was evaluated histologically and histomorphometrically. Four sections of each implant were evaluated; two were located in the cortical and two in the trabecular bone. Of the 44 retrieved implants, 20 implants appeared to be installed partially in the mandibular canal, as clearly visible on the X-rays. These implants were not used in the histomorphometrical measurements. Histological evaluation showed that the trabecular and cortical bone reactions were similar; there was no significant difference in the percentage of bone contact nor in the amount of bone in contact with the implants. In conclusion this study showed that the mandibular corner is an unsatisfactory model for the installation of implants because of anatomical restrictions. Also, the experiment remained inconclusive about the influence of loading conditions on bone behaviour. Nevertheless, the histological results confirmed the bioactive properties of Ca-P coatings. PMID- 15348705 TI - Synthesis and properties of poly(ethylene glycol) macromer/beta-chitosan hydrogels. AB - Semi-interpenetrating polymer network (semi-IPN) hydrogels composed of beta chitosan and poly(ethylene glycol) diacrylate macromer (PEGM) were synthesized and characterized for the application as potential biomedical materials. The mixture of PEGM and beta-chitosan, dissolved in water including a small amount of acetic acid, was cast to prepare hydrogel films, followed by a subsequent crosslinking with 2,2-dimethoxy-2-phenylacetophenone as a non-toxic photoinitiator by ultraviolet irradiation. Photocrosslinked hydrogels exhibited relatively high equilibrium water content in the range 77-83% which is mainly attributed to the free water content rather than to the bound water, hydrogen bonded with components in semi-IPN hydrogels. The crystallinity, thermal properties and mechanical properties of semi-IPN hydrogels were studied. All the photocrosslinked hydrogels revealed a remarkable decrease in crystallinity. The glass transition temperatures, Tg, of crosslinked PEGM segment in semi-IPNs increased compared with poly(ethylene glycol) itself. However, with increasing beta-chitosan content their Tg decreased owing to the higher degree of crosslinking. The tensile strengths of semi-IPNs in dry state were rather high, but those of hydrogels in wet state decreased drastically. PMID- 15348706 TI - In-vitro apatite formation on phosphorylated bamboo. AB - Natural self-reinforced composite, bamboo, was surface modified by phosphorylation with urea-H3PO4 and NaOH-H3PO4 methods; then precalcification was performed by immersing samples in saturated Ca(OH)2 solution. After that, calcium phosphate can be formed on the surface of bamboo samples in calcification media: simulated body fluid (1.5 SBF) and accelerated calcification solution (ACS). Experimental results reveal that pre-calcification is an inevitable step for the formation of calcium phosphate. The calcium phosphate formed in 1.5 SBF was identified by thin-film X-ray diffraction as apatite which was not well crystallized. Compared with the urea-H3PO4 method, the NaOH-H3PO4 method has the advantages of quicker and continuous apatite formation and stronger adhesive between apatite and bamboo. PMID- 15348707 TI - Endothelial cells on plasma-treated segmented-polyurethane: adhesion strength, antithrombogenicity and cultivation in tubes. AB - When the surface of segmented-polyurethane (SPU), where endothelial cells are not capable of proliferating, is modified by plasma treatment, the adhesion and proliferation of bovine aortic endothelial cells (BAECs) can be drastically improved. The cells were capable of proliferating on the inner surface of a plasma-treated SPU-coated tube (length: 50 mm; inner diameter: 1.5 mm). When a steady flow shear stress of 9 Pa was applied to the cells proliferated on the modified SPU surface for 90 min, most cells did not detach from the surface. From an in vitro evaluation test of antithrombogenicity, the cell surface can be considered to provide an inert surface against thrombus formation and blood coagulation. From analyses of the plasma-treated SPU surface, it was suggested that the improvements in BAEC proliferation and adhesion after plasma treatment were due to the change in wettability of the surface. Data suggest that the plasma treatment would be useful for developing a small-calibre hybrid vascular graft. PMID- 15348708 TI - Mechanical properties of carbonated apatite bone mineral substitute: strength, fracture and fatigue behaviour. AB - The synthesis and properties of carbonated apatite materials have received considerable attention due to their importance for medical and dental applications. Such apatites closely resemble the mineral phase of bone, exhibiting superior osteoconductive and osteogenic properties. When formed at physiological temperature they present significant potential for bone repair and fracture fixation. The present study investigates the mechanical properties of a carbonated apatite cancellous bone cement. Flexural strength was measured in three and four point bending, and the fracture toughness and fatigue crack-growth behaviour was measured using chevron and disc-shaped compact tension specimens. The average flexural strength was found to be approximately 0.468 MPa, and the fracture toughness was approximately 0.14 MPa radical m. Fatigue crack-growth rates exhibited a power law dependence on the applied stress intensity range with a crack growth exponent m=17. The fatigue threshold value was found to be approximately 0.085 MPa radical m. The mechanical properties exhibited by the carbonated apatite were found to be similar to those of other brittle cellular foams. Toughness values and fatigue crack-growth thresholds were compared to other brittle foams, bone and ceramic materials. Implications for structural integrity and longer term reliability are discussed. PMID- 15348709 TI - In-vitro development of corneal epithelial cells on a new hydrogel for epikeratoplasty. AB - This study demonstrates the adhesion and growth of bovine corneal epithelial cells on the surface of a new hydrogel. The hydrogel, containing 78% of water and 22% of AN-69 polymer (poly(acrylonitrile-sodium methallyle sulfonate)), was obtained by phase inversion of polymer-dimethylformamide solution in physiological saline (0.9% NaCl). Experiments were also carried out using hydrogel treated with arginine, human albumin and collagen IV. Covering of hydrogel samples by epithelial cells was completed within ten days, with good cell viability. The epithelial cells spread out and formed a consistent cell layer, confirmed by immunocytochemistry experiments against cytokeratins. Transmission electron micrographs showed numerous desmosomes between cells and the presence of some membrane differentiations at the cell/hydrogel interface. This study suggests therefore that the hydrogel might be suitable for the development of artificial epikeratoplasty grafts. PMID- 15348710 TI - A computer-assisted in-vitro biomaterial test for percutaneous devices using human keratinocyte cultures. AB - The aim of this study was to assess the suitability of a computer-assisted in vitro test system to evaluate candidate biomaterials used for percutaneous devices. Silicone rubber (Silastic) and five different polymers from the Eurobiomat concerted action, polyetherurethane, polyvinylchloride with plasticizer, di-ethylhexylphthalate (PVC-DEHP), polyvinylchloride with plasticizer, tri-ethylhexyltrimellitate (PVC-TEHTM), polyethylene and polypropylene were examined with respect to their qualities to facilitate keratinocyte attachment. HaCaT-cells, a spontaneously transformed non-tumorigenic human keratinocyte cell line, were cultured on the different materials for 3 days. Cellular behaviour was examined morphologically by phase-contrast and scanning electron microscopy throughout the 72 h incubation period. For the computer-assisted measurement of the cell-covered substrate surface and subsequent statistical analysis the cells were fixed after 3 days of incubation, stained, photographed and the images then digitally transformed. Of the different polymers examined silicone rubber showed the most favourable results with respect to the quantitative analysis of the cell-covered substrate surface as well as concerning cytomorphological findings. The results of this study indicate that the in vitro test system described is a sensitive screening method for evaluating candidate biomaterials used for percutaneous devices in a fast and reproducible manner, thus reducing number of animal experiments. PMID- 15348711 TI - Characterization of some aromatic dimethacrylates for dental composite applications. AB - Several novel dimethacrylates have been developed as alternative matrix materials for dental composite applications. For the cured bulk polymers the equilibrium water uptake, reduction of glass transition temperatures (Tgs) by water sorption, refractive indices and the surface hardness have been determined. The properties were then compared with those of the control Bis-GMA. These properties correlated well with the structures of the polymers. Polar groups were found to increase the water sorption and thus reduce surface hardness. PMID- 15348712 TI - Influence of Ringer's solution on creep resistance of hydroxyapatite reinforced polyethylene composites. AB - In vitro creep studies of polyethylene, both unfilled and filled with hydroxyapatite at 0.20 and 0.40 volume fraction, have been performed. The samples were immersed in Ringer's solution at 37 degrees C for 1, 7, 30, 90 and 150 days prior to isochronous and creep tests in the same condition. The creep properties of unfilled polyethylene is unaffected by the immersion, but the isochronous modulus and the creep resistance of filled polyethylene were reduced. The effect increased with increasing volume fraction and time of immersion. This reduction is related to the penetration of the solution into the material, softening the interface. PMID- 15348713 TI - Reliability of PMMA bone cement fixation: fracture and fatigue crack-growth behaviour. AB - Fracture mechanics tests were performed to characterize the fracture toughness and fatigue crack-growth behaviour of polymethylmethacrylate (PMMA) bone cement, commonly used in joint replacement surgery. Compact tension specimens of various thicknesses were prepared and tested in both air and Ringer's solution. Contrary to previous reports citing toughness as a single valued parameter, the PMMA was found to exhibit resistance-curve behaviour with a plateau toughness of approximately 0.6 MPa m1/2 in air, and approximately 2.0 MPa m1/2 in Ringer's solution. The increased toughness in Ringer's solution is thought to arise from the plasticizing effect of the environment. Under cyclic loads, the material displayed true mechanical fatigue failure in both environments; fatigue crack growth rates, da/dN, were measured over the range approximately 10(-10) to 10(-6) m/cycle and found to display a power-law dependence on the stress intensity range, DeltaK. The cement was found to be more resistant to fatigue-crack propagation in Ringer's solution than in air. Wear debris was observed on the fatigue fracture surfaces, particularly those produced in air. These findings and the validity of using a linear-elastic fracture mechanics approach for viscoelastic materials are discussed in the context of providing more reliable and fracture-resistant cemented joints. PMID- 15348714 TI - Fixation of carbon fibre-reinforced carbon composite implanted into bone. AB - The push-out test of three types of biomaterials: carbon fibre-reinforced carbon (CFRC), hydroxyapatite (HA), and surgical steel (SS) implanted into rabbits' femurs was carried out. Hydroxyapatite was used as a positive control (good fixation expected in bone) and surgical steel was a negative one (potentially no fixation in bone). Regeneration of bone in contact with all implants was found three months after implantation. The shear strength between CFRC implants and bone was lower than with the HA implants and higher than the shear strength between the surgical steel and bone. Compressive strength of CFRC implants removed after the observation period was significantly lower than the compressive strength of non-implanted samples. It is concluded that the mechanical bonding between the CFRC implants and host tissues exists 3 months after intrabone implantation and is accompanied by a decrease of the strength of implants. PMID- 15348715 TI - Oxygen inhibition of autopolymerization of polymethylmethacrylate-glass fibre composite. AB - The aim of this study was to determine the thickness of the unpolymerized surface layer of autopolymerizing polymethylmethacrylate (PMMA) and PMMA-glass fibre (GF) composite. Powder-to-liquid (P/L) ratios of 10 : 8, 10 : 9 and 10 : 10 by weight of the commercial PMMA was tested and the E-glass fibre weave was used as filler in the PMMA-GF composite. The resin was polymerized between two glass plates at 55 degrees C in air under an air pressure of 300 kPa. Five samples were polymerized for each test group. The inhibition depth was measured by a light microscopic technique with polarized light. The inhibition depth was affected by the P/L ratio of the PMMA: the mean inhibition depth of the unfilled PMMA with the P/L ratio of 10 : 10 was 248.6 microm, while it was 175.4 microm in PMMA with the P/L ratio of 10 : 8 (p=0.044). The inhibition depths were higher in the PMMA GF composite than in the plain PMMA, which was explained by an inadequate impregnation of the GF weave with the PMMA resin. The results suggest that improper impregnation of the fibre product with autopolymerizing PMMA resin can cause oxygen inhibition of the polymerization reaction which should be taken into account when fibre products are clinically used. PMID- 15348716 TI - Mechanical properties of rat epiphyseal cancellous bones studied by indentation testing. AB - Twenty-six pairs of rat femoral heads, distal femurs, proximal tibiae, and humeral heads were tested using an indentation test with a flat-ended cylindrical indentor. A useful mechanical data set including ultimate load, stiffness, and ultimate strength has been generated. Differences were found between the cancellous bones at different locations. Good correlations have been obtained between indentation depth (at 50 N load) and ultimate strength (R=-0.937, p<0.05), which means that with an increase of ultimate strength the indentation depth or deformation decreased proportionally. Based on the experimental results and the comparison with other methods in the literature, the simplicity and usefulness of this indentation test to evaluate compressive mechanical properties of rat epiphyseal cancellous bone are apparent. PMID- 15348717 TI - Multiblock copolyesters as biomaterials: in vitro biocompatibility testing. AB - Cell adhesion, cell growth and cell activities of macrophages and fibroblasts, cultured on newly developed degradable multiblock-copolyesters were studied to examine the biocompatibility and the possible use of these polymers for medical applications. The biocompatibility and the biodegradability of the polymers were confirmed by subcutaneous implantation of polymer foils in rats. The newly developed polymers, two polyesters (DegraPol/bsc43 and DegraPol/bsd43) and a polyesterether (DegraPol/bst41), were found to exhibit good cell compatibility; the cell-to-substrate interactions induced neither cytotoxic effects nor activation of macrophages. The adhesion and growth of fibroblasts and macrophages were different among the substrate. Fibroblasts adhered on the polyesters to about 60% of control cell cultured on tissue culture polystyrene (TCPS) and proliferated in the same doubling time as on TCPS. On the polyetherester cells exhibited weak adhesion; however, they proliferated up to day 4 after plating at the same doubling time as on TCPS (of about 42 h), and then decreased their doubling time to 27 h. Macrophages attached to the polyesters to about 40-60% of TCPS but no significant change was seen in the doubling time of cells cultured on TCPS and the polyesters. Again on the polyetherester, macrophages exhibited relatively low adhesion (25% of TCPS) and high doubling time (about 100 h). Fibroblasts produced high amounts (up to 500% of control cells) of collagen type I and type IV, and fibronectin. Macrophages responded to lipopolysaccharide treatment by the production of nitric oxide (NO) and tumour necrosis factor-alpha (TNF-alpha), indicating that the cell-to-polymer interactions allow fibroblasts and macrophages to maintain their phenotype. All three test polymers exhibit favourable tissue compatibility. The formed capsule was just a few cell layers thick (<30 microm). After 2 months implanted subcutaneously in rats, the molecular weight of the test polymers was reduced by >20% depending on their chemical structure. Taken collectively, the present data demonstrate that the newly developed multiblock copolyesters are biocompatible and biodegradable. PMID- 15348718 TI - Determination of curing time in visible-light-cured composite resins of different thickness by electron paramagnetic resonance. AB - The irradiation time of a visible-light-activated composite necessary to achieve full polymerization throughout the material was studied. Curing-time dependence on the thickness of the material was also investigated. To monitor the visible light-activation effect, the free radical concentration was measured as a function of irradiation time. If the composite sample is less than 0.5 mm thick and exposed to light for a time interval recommended by the manufacturer, full radical concentration is indeed created uniformly. This is not the case in thicker samples. Electron paramagnetic resonance (EPR) was used to monitor the concentration of free radicals in the samples. The number of radicals was monitored as a function of irradiation time during which the radicals were generated in samples 0.5, 0.8, 2.0, 3.0 and 5.0 mm thick. An EPR X-band spectro meter was used to detect the free radical spectra. The number of free radicals per unit mass as a function of irradiation time shows that 60% of the maximum concentration of radicals in a 1 mm sample is reached in 24 s curing time, while in thicker samples it takes hundreds of seconds. On the basis of the experiments, a depth and irradiation time-dependent radical concentration model was developed. This model shows that a 2.0 mm thick sample is cured at the bottom side if irradiated for 60 s. It is proposed that the measure of the degree of polymerization in composite materials should be the polymerization of the bottom layer of the sample which is modelled from the number of free radicals generated in the sample. PMID- 15348719 TI - Solid state synthesis and thermal stability of HAP and HAP - beta-TCP composite ceramic powders. AB - Powders of pure beta-tricalcium phosphate (beta-TCP), hydroxyapatite (HAP) and a biphasic composite mixture of HAP+beta-TCP were prepared by solid state reaction between two commercially available calcium-based precursors namely, tricalcium phosphate (TCP) and calcium hydroxide (Ca(OH)2). These reactants mixed in the molar ratios ranging from 3 : 0 to 3 : 4 (designated T0 to T4) in deionized water, milled and slip-cast into discs were heat treated in the temperature range of 600 degrees C to 1250 degrees C. The products formed were characterized by X ray diffraction (XRD) and i.r. spectroscopic techniques for identification of phases formed and functional groups present in them. While tricalcium phosphate and calcium hydroxide taken in the molar ratio of 3 : 2 and 3 : 3 resulted in pure HAP when heat treated at 1000 degrees C for 8 h, the 3 : 1 and 3 : 1.5 molar ratio compositions resulted in a biphasic mixture of HAP+beta-TCP for similar heat treatments. Heat treatment of 3 : 4 molar ratio composition of tricalcium phosphate and calcium hydroxide at 1000 degrees C yielded HAP with free CaO as the secondary phase. Products of heat treatment at higher temperatures (1150 and 1250 degrees C) for even shorter duration (2 h) while not differing from the products obtained from T0 and T2 cases at 1000 degrees C (pure beta-TCP and pure HAP), change in the case of T1, T1.5, T3 and T4 to products with lesser percentages of HAP containing beta-TCP (in the case of T1 and T1.5) or CaO (in the case of T3 and T4) as the secondary phase. PMID- 15348720 TI - Interactions of human osteoprogenitors with porous ceramic following diffusion chamber implantation in a xenogeneic host. AB - Porous calcium phosphate ceramics are useful bone graft substitutes on account of their osteoconductive properties and lack of toxicity, but they lack osteogenicity and are brittle in nature. Osteogenic properties, and increased biomechanical properties, could be induced by combining them with human bone forming cell populations. Progress has been hampered both by the lack of a suitable experimental assay of in vivo human bone formation and a suitable in vivo test system with which to study such cells in association with biomaterials. Here, trabecular bone-derived cells and marrow stromal fibroblastic cells from four human donors aged between 14 and 27 y have been cultured in vitro then combined with a porous ceramic within diffusion chambers and implanted into athymic mice. Bone and cartilage formation was found within the chambers primed with cells cultured in the continuous presence of dexamethasone and ascorbate. These tissues were found in close apposition to the ceramic, confirming that the material is biocompatible and bioactive. These findings demonstrate both that appropriately primed human-cell populations can express the fully differentiated osteoblastic phenotype in the diffusion-chamber model, and also that this is a useful system in which to test the interactions of such cell populations with putative biomaterials. PMID- 15348721 TI - Highly adhesive hydroxyapatite coatings on titanium alloy formed by ion beam assisted deposition. AB - A compact crystalline hydroxyapatite coating on Ti-6Al-4V substrate with an atomic intermixed coating/substrate interface about 27 nm in width was synthesized by ion beam assisted deposition (IBAD) and a following post treatment. The coating after post-treatment was identified by X-ray diffraction as crystalline hydroxyapatite. The interface between coatings and substrates was studied by Auger electron spectroscopy. The adhesive strength between coatings and substrates was measured by scratch tester. The results showed that the adhesive strength of IBAD coatings is nearly twice that of ion beam sputtered coatings. The study also showed that coatings prepared by IBAD eliminated the interfacial deficiencies existing in plasma-sprayed coatings. PMID- 15348722 TI - Growth of calcium phosphate on phosphorylated chitin fibres. AB - Calcium phosphate growth on chitin phosphorylated fibres was studied using scanning electron microscopy and energy dispersive X-ray analysis (SEM, EDX), micro-Fourier transform infrared spectroscopy (FTIR), and solid state magic angle spinning nuclear magnetic resonance (MAS NMR) techniques. The C6 chemical shift positions of 13C MAS NMR in the chitin fibres phosphorylated using urea and H3PO4 are obvious indicating that phosphorylation takes place not in the C1 but in the C6 region. Micro-FTIR and 31P MAS NMR suggested that ammonium hydrogen phosphate formed during the phosphorylation procedure. Chitin fibres phosphorylated using urea and H3PO4 and then soaked in saturated Ca(OH)2 solution at ambient temperature, which lead to the formation of thin coatings formed by partial hydrolysis of the PO4 functionalities, were found to stimulate the growth of a calcium phosphate coating on their surfaces after soaking in 1.5xSBF solution for as little as one day. The thin layer after Ca(OH)2 treatment functioned as a nucleation layer for further calcium phosphate deposition after soaking in 1.5xSBF solution. EDX-measured Ca : P ratios of the coatings of Ca(OH)2-treated phosphorylated chitin in 1.5xSBF solution suggested that calcium-deficient apatite was formed. PMID- 15348723 TI - Post-irradiation hardness of resin-modified glass ionomer cements and a polyacid modified composite resin. AB - This study examined the post-irradiation hardness of resin-modified glass ionomer cements and a polyacid-modified composite resin using a digital microhardness tester. Change in hardness of these materials over a period of 6 months was compared to that of conventional glass ionomer cements and a composite resin. With the exception of the composite resin, all materials showed a significant increase in hardness over 24 h after their initial set. Dual-cure resin-modified glass ionomer cements showed decreased hardness with increased storage time in saline at 37 degrees C. Results suggest that the addition of resins to glass ionomer cements does not improve initial hardness and does not negate the acid base reaction of conventional cements. Resin addition may, however, lead to increased water sorption and decreased hardness. PMID- 15348724 TI - Fibroblast growth and polymorphonuclear granulocyte activation in the presence of a new biologically active sol-gel glass. AB - The search for chemical devices to be used in clinical orthopaedics must find substances that are biocompatible and do not elicit inflammatory responses in vivo. To this end, a new form of glass has been prepared, composed of 8.1% CaO, 2.9% P2O5, 6.7% N2O5 and 82.3% SiO2, using sol-gel procedures. In order to evaluate the in vitro biocompatibility of this glass, the proliferation of cultured murine fibroblasts and the activation of human polymorphonuclear leukocytes has been studied. The performance of the sol-gel glass has been compared with that of a biocompatible non-resorbable soda-lime glass. Unlike the soda-lime glass, the sol-gel glass neither caused the inhibition of fibroblast growth nor elicited a marked inflammatory response by polymorphonuclear leukocytes, as demonstrated by chemiluminescence assay for reactive oxygen metabolites. PMID- 15348725 TI - XPS characterization of surface films formed on surface-modified implant materials after cell culture. AB - Nitrogen ion-implanted Ti-6Al-4V, Ti-5Al-2.5Fe and 316 L stainless steel and nitrogen or carbon sputter-coated samples were inoculated with rat bone marrow. The interface between the cell layer and the substrata was studied by X-ray photo electron spectrometry and observed by scanning electron microscopy (SEM). Ca and P were detected on all materials after in vitro cell culture. Titanium appears to be present mainly in the form of TiO2. PMID- 15348726 TI - Calcium phosphate formation induced on silica in bamboo. AB - The effect of in vitro induction of calcium phosphate on bamboo surfaces is reported for the first time. Bamboo is studied for biomaterial application due to its elasticity modulus being closer to human bone than other biomaterials. Following an earlier study of cytotoxicity and precipitation of apatite on ground tissue and vascular bundles of bamboo, the composition and function of the minerals in bamboo, especially silica, are considered in the present work. It is found that in both outer and inner surfaces of bamboo culm, there exists some silica. Bamboo elicits an inert response when soaked directly in calcification solution. After the rind of bamboo is treated with sodium hydroxide solution, the silica underneath can induce precipitation of calcium phosphate in an ambient environment. Furthermore, by subsequent grafting with polyethylene glycol (PEG 1000), calcium phosphate induction of bamboo rind can be improved, depending on the concentration of NaOH solution and treatment time. Heat treatment of bamboo can remove the organic materials around the minerals in bamboo, allowing the calcification behaviour of the silica-containing inorganic phase of bamboo in aqueous solution to be studied. PMID- 15348727 TI - Structure and biocompatibility of ion beam modified polyethylene. AB - Structural changes of polyethylene (PE), induced by irradiation with 40 keV Ar+ ions at a fluence of 1x10(12)-1x10(15) cm(-2), are characterized by different experimental methods and physical parameters of the modified PE are related to its biocompatibility. Production of oxidized structures and conjugated double bonds in the PE surface layer modified by the ion irradiation was proved using IR, UV-VIS spectroscopies and a Rutherford backscattering technique. The fusion of macrophages onto implants made of as-irradiated and chemically doped PE was studied in vivo. It was found that the free surface energy is not a decisive factor affecting the non-self-recognition of the modified PE by macrophages. The fusion of macrophages, however, was found to be different on the as-irradiated specimens and the specimens additionally doped with acrylic acid. PMID- 15348728 TI - Porous ceramic lamellae for orthodontic ceramic brackets: part I: fabrication and characterization. AB - This study was undertaken to test a new and original orthodontic bracket base, consisting of a porous lamella, which was designed to facilitate removal of ceramic brackets from the enamel surface after treatment. In the phase of the study presented here, porous pellets were made by bonding coarse alumina particles (calcined or fused) with mullite, formed by their reaction with fine quartz particles during firing of the mixture at 1700 degrees C. After machining the pellets to the desired shape, nine types of lamellae with different porosities were attached to the brackets using two different adhesive resins, and also bonded to bovine enamel using the same adhesives. The tensile bond strengths for the assemblies were determined so that the lamellae and the bonding adhesive that might be suitable for clinical application could be selected for testing in vitro in the second phase of the study. PMID- 15348729 TI - Porous ceramic lamellae for orthodontic ceramic brackets: part II: in vitro performance testing. AB - This study was undertaken to test a new and original orthodontic bracket base, consisting of a porous lamella, which was designed to facilitate removal of ceramic brackets from the enamel surface after treatment. In the phase of the study presented here, two types of lamella and the adhesive resin used to bond them to brackets and teeth, were evaluated in vitro. Two types of test were carried out on bracketed teeth. The tensile bond strength was measured for specimens that had been either kept in water for 24 h at 37 degrees C or subjected to 18000 cycles in water between 6 degrees C and 55 degrees C. The stress required to remove brackets with debracketing pliers was measured and the mode of failure recorded for specimens that had been kept in water for 24 h at 37 degrees C. The results indicate that bracket/lamella assemblies can be bonded to enamel sufficiently strongly for clinical application and can be safely removed without damage to enamel. PMID- 15348730 TI - Optimization of benzoyl peroxide concentration in an experimental bone cement based on poly(methyl methacrylate). AB - The effect of the concentration of benzoyl peroxide in poly(methyl methacrylate) bone cement formulations on their setting characteristics, particularly peak temperature and setting time, were studied. An optimization of the concentration of benzoyl peroxide was made with respect to curing parameters and compared with the residual monomer content. The mechanical properties of the different formulations were also determined and the results indicated that a composition of 1.5% wt/wt and 0.82% wt/wt of benzoyl peroxide and N,N-dimethyl-p-toluidine concentrations, respectively, gave the highest yield strength. Studies on the preparation of bone cement formulations containing different amounts of barium sulphate were also performed to assess the effect on the polymerization process and mechanical properties of the cements. PMID- 15348731 TI - Decomposition analysis of binary polyfunctional urethane monomer mixtures. AB - Thermal decomposition in three binary monomer mixture systems containing an experimentally synthesized monomer (EXP3) and a commercial polyfunctional urethane monomer (U-4TXA) was examined in terms of differential thermal analysis (DTA) and simultaneous thermogravimetry (TG) analysis when a sample was heated to 800 degrees C. The binary EXP3/U-4TXA monomer mixtures were visible light-cured (VLC) resins which included CQ (camphorquinone=0.5 wt%) and DMAEMA (dimethylaminoethyl methacrylate=0.5 wt%) as the photoinitiator. Their DTA curves showed that thermal decomposition initiated at around 300 degrees C and finished at around 500 degrees C. TG curves showed complete weight loss at 800 degrees C during thermally induced decomposition: the decomposition reaction at weight losses of 5 to 50% was calculated from the slope of the (1/T) versus log10(Hr) diagram, because the exothermic decomposition temperature shown on DTA curves increased with increasing heating rate from 2 to 20 degrees C/min. Thermal decomposition analysis showed that a decomposition with weight change had exothermic heats ranging from 0.38 to 1.07 kJ/g for the EXP U1 mixture, 1.06 to 1.76 kJ/g for EXP U2 and 1.74 to 2.02 kJ/g for EXP U3. Activation energy ranged from 1.42 to 1.89 kJ/mol at weight losses of 5 to 50% in the three binary EXP3/U 4TXA monomer mixture systems. PMID- 15348732 TI - Age-hardening associated with precipitation reaction and spinodal decomposition in a commercial dental low-carat Au-Ag-Cu-Pd alloy. AB - Two distinguishable hardening mechanisms, depending on the temperature, were found by isothermal ageing in a commercial dental low-carat Au-Ag-Cu-Pd alloy. Age-hardening was attributed to the precipitation of the metastable AuCu I' and equilibrium AuCu I ordered phases and spinodal decomposition depending on the ageing temperature. It was clearly visible, by using the direct-ageing method, that the XRD peaks of the parent phases showed a shift during transformation. PMID- 15348733 TI - Effect of heat treatment on apatite-forming ability of Ti metal induced by alkali treatment. AB - The present authors previously showed that titanium metal forms a bone-like apatite layer on its surface in a simulated body fluid (SBF), when it has been treated with a NaOH solution to form a sodium titanate hydrogel layer on its surface. This indicates that the NaOH-treated Ti metal bonds to living bone. The gel layer as-formed is, however, mechanically unstable. In the present study, the NaOH-treated Ti metal was heat treated at various temperatures in order to convert the gel layer into a more mechanically stable layer. The gel layer was dehydrated and transformed into an amorphous sodium titanate layer at 400-500 degrees C, fairly densified at 600 degrees C and converted into crystalline sodium titanate and rutile above 700 degrees C. The induction period for the apatite formation on the NaOH-treated Ti metal in SBF increased with the transformation of the surface gel layer by the heat treatment. Ti metal heat treated at 600 degrees C, however, showed a fairly short induction period as well as high mechanical stability, since it was covered with a fairly densified amorphous layer. PMID- 15348734 TI - Glass-ionomer dental restorative: part I: a structural study. AB - A structural study of glass-ionomer cement (GIC) dental restoratives has been completed. Transmission electron microscopy, selected area electron diffraction, and X-ray diffraction studies indicate domain-like microstructure in a new experimental material, whereas a featureless amorphous gel-like microstructure exists in the conventional GIC. Nuclear magnetic resonance studies were also conducted. The new experimental GIC contains domains of (i) bonelike material (apatite), (ii) mesoporous material and (iii) other framework structures (aluminium phosphate in the high cristobalite structure), with its setting chemistry a restructuring of the aluminosilicate glass around the template of poly(acrylic acid). Conventional glass-ionomer cement may set by a similar but slower process. Leaching properties of glass-ionomer cements are also explained. PMID- 15348735 TI - Corrosion of gallium alloys in vivo. AB - The aim of this work was to study the corrosion of gallium alloys in vivo. Three gallium alloys were tested: GF alloy, Galloy and an experimental GaIn alloy. An amalgam was applied as a control. After ageing for a minimum of two weeks, one disc of each of these alloys was mounted with the polished side up in the buccal surfaces of 17 acrylic dentures. Eight sets of the specimens were retrieved after exposure to the oral cavity for 2-4 months, and another seven were retrieved after 6-9 months. Corrosion of the polished cross-sections of the specimens was studied using scanning electron microscopy (SEM). Only the CuGa2 phase was found to corrode substantially in all three of the alloys investigated, leaving behind holes up to 20 microm deep. This is consistent with the corrosion reported after immersion tests in a solution of 0.1 mol lactic acid and 0.1 mol NaCl for 7 days. Such in vitro tests are also reported to cause distinct corrosion of the Sn phase in the gallium alloys. However, a salient feature of the corrosion in vivo was the lack of detectable dissolution of this phase. Thus, for gallium alloys, the accelerated in vitro immersion method produced results which did not agree with clinical observations. Large variations in the corrosion of the CuGa2 were observed from patient to patient. The amount of corrosion on the Galloy specimens appeared to be less and on a finer scale than on specimens of the two other alloys. The depth of corrosion was thus shallower than for this alloy. This finding indicates that there is room for further improvement of the corrosion resistance by modifying the microstructures. Less overall corrosion was found for the amalgam control than for the gallium alloys. PMID- 15348736 TI - Preparation and mechanical properties of calcium phosphate/copoly-L-lactide composites. AB - New artificial bone materials were prepared using calcium phosphates, hydroxyapatite and beta-tricalicum phosphate, and copoly-L-lactide, CPLA. Calcium phosphate powder and CPLA were mixed at 453 K for 10 min with various mixing ratios. Scanning electron microscope observations indicated that the composites of beta-tricalicum phosphate and CPLA were homogeneously dispersed and highly adhesive. Young's modulus of the composites was the same as bone, and bending strength was over half that of bone. The improvement of Young's modulus compared to the original two materials was due to a composite effect. The composites are expected to be usable as artificial bone materials. PMID- 15348737 TI - Electrochemical and surface modifications on N+-ion-implanted 316 L stainless steel. AB - The effect of nitrogen-ion implantation on the electrochemical behaviour of 316 L stainless steel in a simulated physiological solution (HBSS-Hank's Balanced Salt Solution) was studied by open-circuit potential versus time and cyclic polarization techniques, with the aim of characterizing the surfaces and choosing the best nitrogen-ion fluence. Three fluences (10(15), 10(16) and 10(17) ions/cm2) were used. The 10(16) ions/cm2 N+ fluence improves the corrosion resistance of the 316 L stainless steel. PMID- 15348738 TI - Extent of conversion and its effect on the mechanical performance of Bis GMA/PEGDMA-based resins and their composites with continuous glass fibres. AB - The reaction mechanism of bisphenol A bis(2-hydroxy propyl) methacrylate (Bis GMA) and polyethylene glycol dimethacrylate (PEGDMA) was characterized by differential scanning calorimetry (DSC) and infrared (IR) spectroscopy. Composites of S2-glass fibres with different formulations of Bis-GMA/PEGDMA were fabricated by filament winding. The mechanical properties of neat resins and composites were investigated by dynamic mechanical thermal analysis (DMTA) and a three-point-bending flexural test. PMID- 15348739 TI - Comparison of the wear of aged and non-aged ultrahigh molecular weight polyethylene sterilized by gamma irradiation and by gas plasma. AB - The wear of ultrahigh molecular weight polyethylene in artificial joints is a major cause of long-term osteolysis and loosening. The wear rate of aged and non aged ultrahigh molecular weight polyethylene sterilized by both gamma irradiation in air and gas plasma has been studied in simple configuration wear tests. Fourier transform infrared analysis (FTIR) showed marked oxidative degradation of the irradiated and aged material compared to the gas plasma sterilized and aged material. The wear rate of the irradiated and aged material was significantly (three times) higher than the gas plasma sterilized, gas plasma sterilized and aged, and non-sterilized materials. Alternative sterilization procedures such as gas plasma, when used instead of gamma irradiation in air, are likely to reduce the incidence of long-term osteolysis. PMID- 15348740 TI - Morphology and ultrastructure of the interface between hydroxyapatite polyhydroxybutyrate composite implant and bone. AB - A composite of polyhydroxybutyrate (PHB) polymer, reinforced with synthetic hydroxyapatite (HA) particles, with potential as a bone-analogue material, was examined microscopically using scanning electron microscopy and transmission electron microscopy. These imaging techniques provide the means of understanding and monitoring the morphological and structural behaviour of retrieved implants. Scanning electron microscopy was used to assess the overall mechanism of new bone formation at the implant interface after up to 6 months implantation. This procedure was followed by a detailed ultrastructural examination at lattice plane resolution level, using high resolution electron microscopy and selected area diffraction of the regions showing bone apposition. Fine hydroxyapatite crystallites were found to form at the interface after in vivo implantation into cortical bone. PMID- 15348741 TI - Structure property relationship among novel dental composite matrix resins. AB - Several novel dimethacrylate monomers of propoxylated diphenols have been synthesized to establish a correlation between their structure and the important properties of viscosity, curing shrinkage and wetting behaviour. These dimethacrylates were designed to possess linear and flexed structures. These new monomers exhibited very much lower viscosities compared to the conventionally used Bis-GMA. Good correlations were obtained for variations of the monomer structures with bulk viscosities and curing shrinkages. However, the wetting behaviour of these resins did not show any trend. PMID- 15348742 TI - The role of macrophages in bioartificial nerve grafts based on resorbable guiding filament structures. AB - A 10 mm gap in a rat sciatic nerve was bridged by a bioartificial nerve graft consisting of a silicone tube containing seven longitudinally placed filaments made of non-resorbable material (polyamide [Ethilon]) or resorbable materials (polydioxanon [PDS], polyglactin [Vicryl] or catgut). The purpose was to study the tissue reaction induced by the four different types of materials. At 4 weeks an immunocytochemical technique, using ED1 and ED2 monoclonal antibodies, was used to study the presence and location of macrophages. A large number of macrophages were found accumulating on the surface of catgut and polyglactin, while few were found on the surface of polyamide and polydioxanon filaments. It is concluded that the cell layers on the filament surface mainly consisted of ED1 positive cells and their thickness depends on the filament materials. PMID- 15348743 TI - Effects of deproteinization and ashing on site-specific properties of cortical bone. AB - Buffered sodium hypochlorite (NaOCl) solution was used to remove selectively the collagen phase from bovine cortical bone. Changes in the mechanical behaviour and material properties were studied over a wide range of resolution (from 5 microm to 3 mm) using an integrated combination of experimental techniques. Optical microscopy indicated that timed immersion in NaOCl results in cortical bone specimens that consist of a mineralized tissue core surrounded by a layer of deproteinized or anorganic bone. With increased NaOCl treatment, the mechanical behaviour in three-point flexure of the intact specimens became increasingly characteristic of a brittle ceramic material. Localized material properties were evaluated using histology, scanning electron microscopy and microhardness testing. The site-specific properties and the mineralization of the cores were not significantly affected by the treatment; however, the interactions and structural framework of the hydroxyapatite crystallites within the anorganic material were compromised. This destruction of crystallite interlocking was not observed in samples in which the organic phase was removed by ashing at 800 degrees C. The ashed samples maintained microhardness values three times those of the bleached samples. Because of its damaging effects on cortical bone structural integrity, the NaOCl treatment did not provide a reasonable means of studying, as a function of the phasic mass fraction, incremental changes in bone mechanical behaviour or the relative roles of collagen and mineral within the structural hierarchy. PMID- 15348744 TI - Screening of biomedical polymer biocompatibility in NMRI-mice peritoneal cavity: a comparison between ultra-high-molecular-weight polyethylene (UHMW-PE) and polyethyleneterephthalate (PET). AB - The peritoneal resident cell population is influenced by various inflammatory and immunogenic stimuli. The influence of intraperitoneal application of polyethyleneterephthalate (PET) (group A) and ultra-high-molecular-weight polyethylene (UHMW-PE) (group B) powders on peritoneal cell count and macrophage activity was investigated. Powders were tested to mimic wear particles from solid implant devices as these particles often cause chronic granulomatous inflammation. The results were compared with the inflammatory response following an abdominal midline incision (group C) and untreated animals (group D). On days 1, 7, 14 and 30 peritoneal cells were quantified and the number of active macrophages was assessed. Groups A and C mice showed a significant loss of macrophages in the peritoneal lavage at day 1 but this returned to normal values (group D) on day 7. In contrast, group B animals remained at low peritoneal cell counts but showed the highest number of active macrophages. Only in this latter group was adhesion formation and granulomatous clustering of polymer powder observed. Applying the parameters macrophage count and the number of active macrophages it can be concluded that PET elicits a weaker inflammatory reaction than UHMW-PE in mice peritoneal cavity. Thus this animal model may be used as a screening test for biomedical materials, especially their wear products. PMID- 15348745 TI - Crystal morphology and texture in calcium oxalate monohydrate renal calculi. AB - The morphology of calcium oxalate monohydrate crystals in renal calculi has been studied. Morphology modifications with respect to equilibrium morphology have been explained from the structure of (1 0 0) crystal faces and the effect of polyelectrolytes present in urine. The texture of the stones has been considered as the result of both primary aggregation and crystal aggregation produced by anionic macromolecules. The arrangement in concentric layers of crystals with a radial disposition has been related to a process of intense nucleation induced by anionic macromolecules and typical polycrystalline growth on films. PMID- 15348746 TI - Morphological and chemical characterization of microfabricated fibres for biological applications. AB - Monodisperse fibres and particulates of different materials with controllable three-dimensional shape, size and chemical composition are of interest in research on toxic respirable fibres as well as wear debris around orthopaedic implants. We have previously demonstrated the production of well-controlled, metal and oxide microfabricated fibres having dimensions 0.1 to 10 microm. While our previous results focused on how controlled fibres can be prepared by microfabrication methods, this paper evaluates property-production relationships for microfabricated fibres. Here we have briefly reviewed the production of 0.1 microm x 0.5 microm x 10 microm microfabricated fibres made by electron beam lithography from evaporated titanium or silicon oxide films using a double lift off method. We have also analysed the properties of these fibres with respect to morphology and chemical composition, and how they are affected by variations in the production process. Two different solution types have been used to place fibres into liquid suspension and to clean and sterilize them for biological testing. One method involves the use of organic solvents; the other a hydroxide solution and water. While fibre dimensions appear to be material-specific, differences can be corrected for by compensation of the size of the lithographic pattern. Similarly the crystallinity of fibres is material-specific, as is to be expected of evaporated thin films, but should be possible to modify by varying deposition parameters or heat treating, for example. Of the cleaning methods used, the one using an aqueous hydroxide solution is preferred over solvent cleaning, as it is easier to perform and appears to be more effective at removing resist from the fibre suspension. PMID- 15348747 TI - Biocompatibility of solid poly (ortho ester). AB - In previous studies poly (ortho ester) (POE) has shown promise as a resorbable device, a hemostatic sealant and as a carrier for drugs in bone surgery. The aim of this study was to evaluate the tissue reactions of solid poly (ortho ester) implanted into both tibiae of 17 rabbits. One half of the rods were sterilized by gamma radiation and the other half by ethylene oxide. The follow-up times were from 1 week to 21 weeks, after which the animals were killed and the bony specimens examined histologically. The connective tissue samples were examined immunohistochemically in order to study the occurrences of two extracellular matrix glycoproteins, tenascin and fibronectin. The results showed that solid poly (ortho ester)s induce a moderate inflammatory reaction for 9 weeks. Tenascin and fibronectin were present in samples from 1 week up to 4 weeks. It was also found that gamma sterilized POE was resorbed at week 7 and ethylene oxide sterilized POE at week 13. PMID- 15348748 TI - Infrared and Raman microspectrometry study of fluor-fluor-hydroxy and hydroxy apatite powders. AB - Visible Raman and infrared microspectrometry studies performed on fluorapatite and hydroxyapatite powders have shown similar results. Small modifications of the nu2 and nu4 PO(3-)4 tetrahedra bending modes are observed. A small frequency shift of the nu1 mode and modifications on the nu3 mode region accompanied with a simplification of the hydroxyapatite and fluorapatite respective spectra from seven to four bands were observed. A broad and weak band which could be attributed to the Ca-F bond is detected at 311 cm(-1) on the Raman fluorapatite spectra. The phosphate vibration modes are little disturbed by fluoride substitution. This could indicate that phosphate groups interact strongly between themselves and weakly with substituted atoms (i.e. hydroxyle and fluoride atoms). Whatever crystallographic model is considered, the number of bands observed is always lower than the number of calculated ones, even for hydroxyapatite, whose symmetry is lower than that of fluorapatite. PMID- 15348749 TI - Ageing reactions in a high carat gold alloy for dental porcelain bonding. AB - The phase transformation during continuous heating of a high carat gold alloy used for porcelain bonding was investigated by electrical resistivity measurements, hardness tests, X-ray diffraction and scanning and transmission electron microscopy. Four reaction stages (I, II, III and IV) were found. Stage I corresponded to the formation of a short-range order. A discontinuous precipitation took place in stage II, which contributed to remarkable hardening. Stages III and IV were reactions to the stable phases at each temperature region, and resulted in softening. The activation energies for stages I, II and III are 27.1, 33.8 and 58.2 kcal/mol, respectively. PMID- 15348750 TI - Influence of hydrogen bonding on properties of BIS-GMA analogues. AB - The influence of chemical structure on the important properties of composite matrix resins is being systematically investigated. This study addresses the relationships between pendent side chain structures, viscosity and curing shrinkage. In particular, viscosity is known to be greatly influenced by intermolecular interactions, such as hydrogen bonding, and free volume effects. In order to establish the relative importance of these factors, analogues of BIS GMA were synthesized in which the pendent hydroxyl groups were replaced by trimethyl siloxyl, and by dimethyl, isopropyl siloxyl groups. The viscosities were determined with a cone and plate viscometer and curing shrinkages were determined gravimetrically. They were compared to previously determined values for BIS-GMA and its methyl and hydrogen substituted analogues. The high viscosity of BIS-GMA is drastically reduced by replacement of the hydroxyl group, or its substitution by silylation. The relatively smaller effects produced by varying the bulk of the substituted side chains indicates that the main effect on viscosity is due to the presence or absence of hydrogen bonding. Conversely, increasing the bulk of the side chain substituent has less effect on viscosity, but significantly reduces the curing shrinkage. Changes in curing shrinkages were explained in terms of effects of free volumes associated with the monomers. PMID- 15348751 TI - Ex vivo complement protein adsorption on positively and negatively charged cellulose dialyser membranes. AB - An ex vivo test system was used to measure complement protein C3 and factor B adsorption onto small dialyser modules made from regenerated and modified cellulosic hollow fibre membranes in which positive diethylaminoethyl (DEAE) or negative carboxymethyl (CM) groups were introduced into the cellulose matrix. The extracorporeal system, which included test-dialysers and the dialysis environment, allowed the use of labelled proteins without contaminating the blood donors which were connected in an open-loop fashion to the extracorporeal test system. The modules were removed at selected time points from the extracorporeal system for radioactivity counting. The results were used to evaluate the mechanisms involved in complement reactions to foreign surfaces. The system therefore allowed the analysis of complement protein adsorption occurring in the dialyser modules and its relationship to the complement generation rate in the extracorporeal system to be evaluated. It was possible to demonstrate that significant complement C3 and factor B adsorption occurred in the test modules made of cellulosic membranes. Complement adsorption as a function of the pH and the release reaction of the adsorbed C3 and factor B after membrane blood perfusion were therefore found to be variable according to the cellulosic membrane type and the presence of positive or negative charged groups within the cellulose matrix. The data obtained from the ex vivo model therefore provided additional evidence on the discussion of the mechanisms involved in the increased complement activation by regenerated cellulose and in its attenuation by DEAE- or CM-modified cellulose. PMID- 15348752 TI - Fibrous growth of tricalcium phosphate ceramics. AB - Structural transformation and sintering processes of tricalcium phosphate (TCP) ceramics prepared from defective hydroxyapatite (Ca9HPO4(PO4)5OH) were studied by X-ray diffraction (XRD) and atomic force microscopy (AFM). Starting powders with Ca/P ratio approximately 1.5 were obtained by adding 0.5 l of 0.3 M H3PO4 solution to an equal volume of 0.45 M Ca(OH)2. In the prepared ceramics, the onset temperature for transformation of defective hydroxyapatite into TCP (witlokite) agrees with the onset temperature for sintering (800 degrees C). Sintering occurs through the formation of a fibrous structure, which resembles biological hard tissue. In the 1000-1200 degrees C range, these fibres coalesce into grains of up to 0.6 microm in size with a fibrous-laminar morphology. At the end of this sintering stage witlokite transforms into alphaTCP. At about 1450 degrees C, partial decomposition of alphaTCP into Ca2P2O7+Ca4P2O9 is observed. AFM observations suggest that Ca2P2O7 is segregated in the liquid state and increases the velocity of grain growth (up to 12 microm). PMID- 15348753 TI - Surface degradation features and microstructural properties of ultra-high molecular weight polyethylene (UHMWPe). AB - Surface degradation of UHMWPe is recognized as a leading clinical concern, limiting the long-term performance in total knee replacements. Eight retrieved tibial plateaux and six wear screening test samples were evaluated for surface degradation features and microstructural features. The surface degradation features were assessed using stereomicroscopy and scanning electron microscopy. Microstructural features were evaluated using optical microscopy of thin cross sections and a permanganate etching technique. The pitting mechanism of wear was observed on all eight retrieved TKR and covered an average of 12.6% of the surface area. The size of the pits were similar to the size of grains observed in the microstructural evaluation - approximately 100 to 200 microm. The presented observations of pitting in retrieved knee implants have shown that the post processing microstructure may influence this mechanism of surface degradation and hence the wear products. PMID- 15348754 TI - Strength retention of self-reinforced poly-L-lactide screws. A comparison of compression moulded and machine cut screws. AB - The effect of the manufacturing method on the strength retention of self reinforced poly-L-lactide (SR-PLLA) screws was studied in vitro and in vivo from 3 up to at least 15 weeks. SR-PLLA screws were manufactured from axially oriented SR-PLLA billets by the conventional compression moulding process and an in-house developed machine cutting technique. New machined SR-PLLA screws (thread diameters 4.5 mm and 3.5 mm) were significantly stronger than older compression moulded SR-PLLA screws (4.5 mm and 3.5 mm) in bending and torque strength tests but significantly weaker in shear strength tests. In pull out tests there were not significant strength differences between the screws. Mechanical analysis and molecular weight measurements confirmed earlier observations that SR-PLLA degrades faster in vivo than in vitro. These results suggest that the new screws could be suitable for clinical use. PMID- 15348755 TI - Microcapsules prepared from starch derivatives. AB - In this work, vinyl groups were introduced on soluble starch by reaction with 2 vinyl-4,4-dimethyl-2-oxazolin-5-one. The polysaccharides obtained were characterized by 1H-NMR and DSC. The 1H-NMR spectra showed high degrees of substitution and the DSC thermograms suggest a low crystallinity in the modified starch. The modified starch was used to obtain microcapsules prepared through interfacial crosslinking with DPGDA (dipropyleneglycol diacrylate) by a water-in oil emulsion polymerization. PMID- 15348756 TI - Physico-chemistry and cytotoxicity of ceramics: part I: characterization of ceramic powders. AB - The morphology of Al2O3, ZrO2/Y2O3, AIN, B4C, BN, SiC, Si3N4, TiB2, TiC, TiN ceramic, graphite and diamond powders has been studied by scanning electron microscopy (SEM) and the specific area of each powder was determined with the BET method. X-ray diffraction (XRD) investigations have been carried out in order to evaluate the crystallinity and determine the constitutive phases. The chemical composition was assessed by classical chemical analyses and by X-ray microprobe; some powders were studied by the laser micro-Raman technique. Correlations have been established between all these results. PMID- 15348757 TI - Investigation into cell growth on collagen/chondroitin-6-sulphate gels: the effect of crosslinking agents and diamines. AB - Artificial skin substitutes based on cultured autologous keratinocytes need to have sufficient strength and ease of handling to be utilized successfully by surgeons in the clinic. This may be achieved by crosslinking the collagen substratum on which the cells are cultured, which in this case is a collagen gel. Increased strength must be attained without detrimental effect on cell growth. The influence of potential crosslinking agents including the glycosaminoglycan, chondroitin-6-sulphate (Ch6SO4), the water soluble carbodiimide crosslinking agents 1-ethyl-3-(3-diaminopropyl) carbodiimide (EDAC), and 1,1 carbonyldiimidazole (CDI), and the polyamines putrescine, spermine and diaminohexane, on cell growth rate has been investigated. Incorporation of 20% Ch6SO4 into collagen gels caused an approximately 16% increase in keratinocyte growth, but had no significant effect on that of dermal fibroblasts. Pre-formed collagen gels (+/- Ch6SO4) were treated with the carbodiimides. This crosslinking treatment markedly inhibited fibroblast growth (EDAC 45% inhibition, CDI 70%), without affecting that of keratinocytes. Pre-formed collagen gels (+/-Ch6SO4 and carbodiimide) were treated with 0.1 M, 0.5 M or 1.0 M polyamine. Spermine inhibited the growth rate of both cell types at all concentrations tested, whereas putrescine and diaminohexane had little effect. The mechanical strength of these crosslinked gels is currently being assessed to determine the optimum composition in terms of cell growth and biocompatibility, and strength. PMID- 15348758 TI - Preparation and characterization of fluoride-substituted apatites. AB - Apatites were prepared with three different fluoride concentrations: 0.0 mM (pure hydroxyapatite) 2.5 mM and 5 mM. Reactions were performed in aqueous medium using a reaction between diammonium orthophosphate and calcium nitrate 4-hydrate and ammonium fluoride at temperatures of 3 degrees, 25 degrees, 60 degrees and 90 degrees C. The effects of reaction temperature and fluoride concentration on the crystal morphology, phase purity and crystallinity of the precipitates were observed, using transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy and ion chromatography. Transmission electron micrographs revealed that the crystallites precipitated at 3 degrees C were spheroidal, but became increasingly acicular with increasing precipitation temperature. X-ray diffraction results indicated that all the materials produced were phase pure and that the crystallinity of apatites prepared at higher precipitation temperatures was higher than those prepared at lower precipitation temperatures. A significant difference in the a-axis dimension of fluoride-substituted apatites was observed, as compared to hydroxyapatite. FTIR spectroscopy revealed a hydroxyl band at 3568 cm-1, along with a broad peak of adsorbed water in the region of 3568 cm-1 to 2670 cm-1 in the hydroxyapatite and fluoride-substituted apatite powders. Hence by careful selection of the precipitation conditions and fluoride contents, the composition and morphology of fluoride-substituted apatite may be controlled and this has interesting implications for the development of these materials for biomedical implantation. PMID- 15348759 TI - Plasma spraying of zirconia-reinforced hydroxyapatite composite coatings on titanium: part I: phase, microstructure and bonding strength. AB - Plasma-sprayed hydroxyapatite (HA) coatings applied to metal substrates can induce a direct chemical bond with bone and hence achieve biological fixation of the implant. However, the poor bonding strength between HA and substrate has been of concern to orthopaedists. In this study, two submicrometre ZrO2 powders stabilized with both 3 and 8 mol% Y2O3 (TZ3Y and TZ8Y, respectively) were incorporated in a plasma-sprayed HA coating on Ti-6Al-4V substrate to investigate the change in phase, microstructure and bonding strength. The results show that ZrO2 composite coatings contain more unmelted particles and greater porosity. During plasma spraying, ZrO2 reacts with the CaO in HA to form CaZrO3 and accelerates HA decomposition to alpha-TCP and Ca4P2O9. Nevertheless, bonding strength increases with increase of ZrO2 content in the range 0 to 10 wt% studied. The higher Y2O3-containing TZ8Y apparently exerts a greater strengthening effect than the lower Y2O3-containing TZ3Y. PMID- 15348760 TI - Plasma spraying of zirconia-reinforced hydroxyapatite composite coatings on titanium: part II: dissolution behaviour in simulated body fluid and bonding degradation. AB - The change of phase, morphology and bond strength of plasma sprayed hydroxyapatite (HA) coating and ZrO2/HA composite coatings immersed in simulated body fluid (SBF) for various periods of time was studied. X-ray diffractometry (XRD) and scanning electron microscopy (SEM) were used to identify the phase and observe the morphology of the coating surface before and after immersion. In addition, inductively coupled plasma emission spectroscopy (ICP) was used to measure the ion release rate of coatings in SBF for various periods of time. Observation of the morphology by SEM shows that the composite coating with the addition of ZrO2 in HA significantly reduced the dissolution rate of impurity phases in simulated body fluid. The argument was supported by measurement of Ca2+ ion concentration in SBF. During plasma spraying, less OH- ions were lost in a ZrO2-containing composite coating. This factor, together with the reduced effective surface of the ZrO2-containing HA coating, were attributed to the reduced dissolution rate of the composite coatings. All the plasma sprayed coatings degraded after immersion in SBF owing to dissolution of constituents in the coating, however, the addition of ZrO2 in HA improved the bonding strength of HA coating after immersion in SBF. PMID- 15348761 TI - Electrophoretic deposition of hydroxyapatite. AB - Hydroxyapatite powders were prepared by a chemical precipitation method and electrophoretically deposited on Ti6Al4V surgical alloy substrates. The powders were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), particle size distribution and zeta potential measurements. Prior to electrophoretic deposition, anodic films were obtained on Ti6Al4V and studied by the Auger method. It was established that experimental conditions of powder preparation, electric field and stirring have a significant influence on suspension stability and deposit morphology. The deposition yield was studied at various deposition durations and applied voltages. Sintered coatings were studied by SEM and XRD. PMID- 15348762 TI - Modelling the impact testing of prescription lenses. AB - Lenses are tested in an impact test in which a steel ball is dropped from a height onto the centre of the lens. This causes the lens to deform until the stress in the lens reaches a point at which fracture occurs. A survey of the literature was carried out and analytical models of the load/deflection and of the deflection/stress relationships were selected. A mathematical model of the impact test on lenses was developed. This model consisted of calculating the load deflection relationship of a lens loaded at a central point, combined with calculating the deflection at which fracture occurred. From this model the impact energy required to deform a lens to fracture was obtained. This was held to be equal to the minimum kinetic energy of an impactor, less losses, that would be needed to cause lens fracture. As the losses are small, the calculated energy was used as an estimate of the impact strength of the lens. These values were then compared to those established by experiment. The impact energies predicted by the model were a close approximation of the experimental results for the lenses tested. PMID- 15348763 TI - Fabrication of hydroxyapatite ceramic with controlled porosity. AB - Porous hydroxyapatite (HAp) ceramics were fabricated using poly vinyl butyral (PVB) as a porosifier. The effects of preparation conditions involving PVB particle size, sintering time, and forming pressure (die-pressing technique) on the resultant pore size/structure as well as the pore size distribution were investigated. The experimental results showed that the HAp ceramics with controlled pore characteristics such as pore volume fraction, pore size and pore structure are achievable. It provides the possibility to design HAp ceramics with diverse porosities simulating that of natural bone. PMID- 15348764 TI - The use of rat, rabbit or human bone marrow derived cells for cytocompatibility evaluation of metallic elements. AB - Rat, rabbit and human bone marrow cells were cultured according to the method previously reported for cells of rat origin [1] and were exposed, or not (control), to corrosion products of a Co-Cr orthopaedic alloy as well as to metal salts containing Co2+, Cr3+ and Cr6+. Cells were cultured for 21 days and analysed for the following biochemical parameters: intracellular MTT reduction (i.e. cell viability/proliferation), alkaline phosphatase (ALP) activity and protein production. Morphological observations included both histochemistry (detection of ALP-positive cells, calcium and phosphate deposits) and scanning electron microscopy (SEM). Control cultures of rat and rabbit cells showed higher proliferation rates than human cells at the start of culture, but they all reached similar values on day 21. Protein production was parallel to cell proliferation. In contrast, ALP activity of rat cultures was much stronger than rabbit or human cultures. All cell types were able to develop the osteogenic phenotype in vitro.Co-Cr extract caused inhibitory effects on cell viability, on ALP activity and, to a lower extent, on protein production of all rat, rabbit and human cell cultures. Compared to rat and rabbit cultures, human cultures were the most sensitive to metal ions exposure. PMID- 15348765 TI - Dynamic blood cell contact with biomaterials: validation of a flow chamber system according to international standards. AB - The increasing number of patients requiring prosthetic substitution of segments of the vascular system strongly supports the need to optimize a relevant, standardized testing panel for new materials designed for synthetic vascular prostheses. The ISO gives the standard requirements for testing biomaterials provided for implantation. Our primary interest was the establishment of a reliable in vitro panel as a useful and relevant screening system for vascular implant devices to evaluate blood/device interactions under flow conditions. The aim of the present study was to evaluate influences of different flow conditions on blood cell-biomaterial interactions with special emphasis on the interactions of human granulocytes (PMN) and polymeric surfaces. PMN were isolated and vital cells were quantified by flow cytometrical analysis directly before, as well as immediately after the experiments. The viscosity of the final cellular suspension was analysed by using a computerized cone-plate rheometer. As reference materials we used FEP-teflon, PVC-DEHD, PU, PP and PE. Dacron and ePTFE synthetic vascular protheses were tested in a comparative way to those references. The adhesion processes were observed over a period of 40 minutes under arterial (shear stress 0.74 Pa) and venous (shear stress 0.16 Pa) flow conditions in a parallel plate flow chamber system under highly standardized conditions and laminar flow. The cells were observed with the help of inverse light microscopy. Cell behaviour was recorded and analysed in both analogue (video) and digital (imaging system) modes. Samples of the cell suspensions were obtained at regular time intervals and analysed by enzyme linked immuno sorbent assay (ELISA) to quantify LTB4 release. Irrespective of the material, approximately 3 to 4 times more PMN adhered to the biomaterial surfaces under venous flow conditions compared to the arterial. Shear intensity did not influence the running order of biomaterials with respect to cell numbers. This response in descending order at the end of the experiments was as follows: PU, PVC-DEHD, PP, PE and ePTFE. The biochemical analyses indicate that in the system used only a weak effect on LTB4 release induced by the different materials could be determined. A significant effect caused by flow conditions was not observed. Further experiments, both static as well as dynamic, must be performed for multiple, relevant parameters of haemocompatibility, for potential biomaterials as well as those currently in use in vascular prostheses. PMID- 15348766 TI - The cell and molecular biological approach to biomaterial research: a perspective. AB - The past two decades have witnessed a revolution in our understanding of chemical processes in living organisms. This is mainly a result of the massive advances in the fields of cell and molecular biology. These techniques are highly relevant to the biomaterials sector, as they offer the scientist the possibility to better understand the mechanisms involved in the interactions between cells and a material surface-a prerequisite for the rational development of medical devices with optimal biocompatibility. The purpose of the present article is to explain the rationale of the cell and molecular biological approach to biomaterial research and to present typical examples from the authors' laboratory, as well as from the literature, to illustrate its application. Important aspects of interfacial biology, including the underlying biological mechanisms and methodology, are presented. Of the latter the combination of morphological techniques with methods of cell and molecular biology as well as molecular genetics (so-called "combinative techniques") are particularly useful. The applicability of this approach is illustrated from a study on the pathomechanisms of metal ion-induced inflammation. In addition, the approach is essential to the development of targeted intervention strategies, as for example in the luminal surface modification of vascular prostheses to permit endothelial cell seeding. PMID- 15348767 TI - A possibility for new evaluating method of cytotoxicity by using heat shock protein assay. AB - To determine whether heat shock proteins can be utilized as a biomarker for cytotoxicity of dental materials the induction of synthesis of heat shock proteins by mercuric chloride was examined. To analyse the synthesis of heat shock proteins, HeLa cells were labelled with [35S] methionine, and the labelled proteins were separated by SDS-PAGE and autoradiographed. Incubation of the cells in a medium containing 1.25 to 40 microM mercuric chloride markedly increased the synthesis of HSP70. At 20 or 40 microM mercuric chloride in medium, HeLa cells synthesized HSP70 at 2 h after exposure, maximally at 4-7 h, and gradually diminished thereafter. Examination of the cytotoxicity of mercuric chloride by the conventional neutral red uptake assay revealed a reduction of uptake of the dye in the presence of mercuric chloride at 40 microM and above. These findings suggest that the induction of synthesis of HSP70 is one of the most sensitive cellular responses caused by mercury ion, and the heat shock protein assay can be utilized for evaluation of the cytotoxicity of dental materials. PMID- 15348768 TI - Dental application of binary urethane monomer mixtures:strengthened resin matrix. AB - The strengthened resin matrices in six experimental binary visible light-cured (VLC) urethane monomer mixtures were examined in terms of mechanical strength. A 60 wt% bis-GMA/40 wt% binary monomer mixture was used as a control sample. A dry state (1 day in air at 37 degrees C) and a wet state by immersion in distilled water (7 and 30 days at 37 degrees C) were investigated. The compressive strength and diametral tensile strength of urethane monomer samples had values equivalent to, or greater than those of a control sample in the dry state, but had increased values in the wet state. The nano-indentation hardness values in experimental urethane monomer mixtures were greater than that in a control sample in the dry state, showing that immersion in water gave increased strength (maximum increase was about 2.5 times) in six binary urethane monomer mixtures. The compressive strength of the samples in the dry and wet states exhibited linearly increasing elastic modulus values for the resin matrix in the range about 50 to 2600 MPa. The toughened resin matrix had smaller amounts of residual monomer in the urethane monomer mixtures (0.41 to 5.03 wt%) compared with that of the binary 60bis-GMA/40TEGDMA mixture (0.71 to 6.26 wt%). This study has revealed that resin matrices are strengthened by the use of four-functional urethane monomers in experimental binary urethane monomer mixtures. PMID- 15348769 TI - Bioactivity of sol-gel derived organically modified silicates: part i: in vitro examination. AB - Bioactivity was investigated for several organically modified silicates (Ormosils) prepared through sol-gel processes. Ca(II)-free samples were biocompatible only, but Ca(II)containing samples were bioactive and deposited apatite during immersion in a simulated body fluid. The ease of silanol (Si-OH) group formation on the ormosils was considered a predominant factor controlling the bioactivity, while the effect of dissolved Ca(II) ions to increase the degree of supersaturation in the simulated body fluid is secondary. PMID- 15348770 TI - Polyamide 4,6 membranes for the encapsulation of Langerhans islets: preparation, physico-chemical properties and biocompatibility studies. AB - Porous polyamide 4,6 membranes developed as semi-permeable and biocompatible membranes for the encapsulation of pancreatic islets were prepared by precipitation in water. Different membrane morphologies were obtained by varying the molecular weight of polyamide 4,6, the solvent evaporation time and the composition of the casting solution. Each membrane was submitted to differential scanning calorimetry and water flux measurements to study the total water content and the different kinds of water in correlation with its permeability performances. Their biocompatibility was first evaluated by a surface protein adsorption test. Of the various membranes, the one prepared by immersion in water after 5 minutes of solvent evaporation, of 15% KS200 polyamide 4,6 solution added with 1% of polyvinylpyrrolidone K30 seems to be the most promising. This membrane is characterized by a low adsorption of proteins, a high hydraulic coefficient and an asymmetric structure. Such a membrane represents a good candidate to be an efficient immunological barrier. It also exhibits good glucose and insulin diffusion properties. Moreover, rat islets cultivated on its surface were not affected by its presence and no important cell adhesion was noticed when implanted in mice. This membrane exhibits most of the properties suitable for the islet encapsulation with a view to developing a bioartificial pancreas. PMID- 15348771 TI - Non-destructive characterization of hydrogels. AB - Hydrogels have been prepared by a freezing-thawing procedure and investigations made of the effect of both number of freezing-thawing cycles and different content of hyaluronic acid (HA) on the mechanical properties of the PVA-HA hydrogels using non-destructive testing. The bulk elastic modulus K of hydrogels has been determined by pulse-echo measurements. It is noted that hydrogel elastic properties improve with the number of the cycles in PVA-HA 100/0; on the other hand samples with a high HA (1,000,000 molecular weight) content, beyond the third cycle, seem to be unaffected by the number of cycles. A bulk elastic modulus fall-off is then observed in samples submitted to an additional overnight freezing between two subsequent cycles. K increases in hydrogels with the highest HA content, when samples undergo pulse-echo measurements soon after their preparation. When hydrogels reach equilibrium, after having been kept in deionized water for 12 h, K values are lower, showing a nearly constant behaviour with different PVA-HA ratios and cycles. Furthermore, by means of scanning laser acoustic microscopy (SLAM) defects have been detected in the hydrogels. In samples which have reached equilibrium, SLAM images show that these defects disappear in PVA-HA hydrogels. PMID- 15348772 TI - Mineralization in serially passaged human alveolar bone cells. AB - Well-characterized human bone cell cultures have been regarded as a useful tool to study bone control mechanisms and also to analyse bone/biomaterials interactions. In the present study, human alveolar bone cells were cultured in alpha-minimal essential medium (alpha-MEM) containing 10% foetal bovine serum (FBS), 50 microg/ml ascorbic acid, 10 mM sodium beta-glycerophosphate and either in the presence or in the absence of 10 nM dexamethasone (Dexa). Cultures were characterized concerning cell viability/proliferation, alkaline phosphatase (ALP), acid phosphatase (ACP) and tartraric acid resistant phosphatase (TRAP) activities, and formation of mineralized areas. Cell proliferation increased gradually for approximately 20 days. In the presence of Dexa, cells formed isolated or interconnected multilayered clusters that increased with culture time. Histochemical assays revealed strong positive reactions for ALP and calcium and phosphates deposits, mainly in relation t! o ce lls associated with the clusters. High levels of ALP activity (biochemical determination) were observed. Cells cultured in the absence of Dexa showed significantly lower ALP activity and no calcium and phosphates deposits were present. Serially passaged cells kept the proliferation rate constant but a decrease in ALP activity was observed either in the presence or in the absence of Dexa. The ability to form mineralized areas (cultures fed with Dexa) also decreased on serial subculture. PMID- 15348773 TI - Ovine anterior cruciate ligament reconstruction using a synthetic prosthesis and a collagen inductor. AB - The evaluation of composite anterior cruciate ligament prostheses, the union of an inductor of collagen and a synthetic fibre, are described. They were implanted in 10 sheep models for six months. None of the ligaments were broken, and the dynamic radiography was stable during this time. After harvesting them, a histologic study was performed on the intra-osseou and intra-articular portion, and on the synovial tissue. Biocompatibility was excellent. An osseous anchorage was found in 50% of cases and a fibrous ingrowth with well oriented fibres in each case. The results of this study show that the matrix has only partially played its role, because there was no improvement of fibrous ingrowth compared with other studies. However this fact corresponds to results at only 6 months and it is necessary to identify the type of collagen. PMID- 15348774 TI - In-situ polymerization behaviour of bone cements. AB - The polymerization behaviour of bone cements during total hip replacements is characterized by a fast and highly non-isothermal bulk reaction. In the first part of this paper the reaction kinetics are analysed by calorimetric analysis in order to determine the rates of polymerization in isothermal and non-isothermal conditions. A phenomenological kinetic model, accounting for the effects of autoacceleration and vitrification, is presented. This model, integrated with an energy balance, is capable of predicting the temperature across the prosthesis, the cement and the bone and the degree of reaction in the cement, during in situ polymerization. The temperature and the degree of reaction profiles are calculated, as a function of the setting time, taking into account the system geometry, the thermal diffusivity of bone, prosthesis and cement, and the heat rate generated by the reaction according to the kinetic model. Material properties, boundary and initial conditions are the input data of the heat transfer model. Kinetic and heat transfer models are coupled and a numerical solution method is used. The model is applied in order to study the effects of different application procedures on temperature and degree of reaction profiles across the bone-cement-prosthesis system. PMID- 15348775 TI - An experimental study in X-ray spectroscopy of the zirconium (Ca-PSZ) - bone interface. Microanalytic evaluation of the osteogenetic response. AB - Ultrastructural difractometric and chemical evaluations of calcium partially stabilized zirconium (Ca-PSZ) implants were performed in an in vivo study on animals in order to evaluate its biological behaviour. The chemical-morphological investigations demonstrated the presence of an osteogenetic activity at the bone biomaterial interface. The new-osteogenesis was preceded by the formation of a loose connective tissue around the implants. This mesenchymal-type tissue without a capsular organization, allowing modulation of the mechanical forces to which the implant is subject, could be considered a positive event in the osteogenetic process and not a sign of future failure of the implant. Finally, microanalytical investigations carried out on non-implanted and implanted Ca-PSZ tools suggested that the surface of this ceramic material does not undergo modification once it has been inserted in the biological environment (12 months). PMID- 15348776 TI - An investigation of the chemical synthesis and high-temperature sintering behaviour of calcium hydroxyapatite (HA) and tricalcium phosphate (TCP) bioceramics. AB - The experimental conditions for the synthesis of sub-micrometre, spherical particles of calcium hydroxyapatite [Ca10(PO4)6(OH)2] (HA) and tricalcium phosphate [Ca3(PO4)2] (TCP) are investigated through chemical coprecipitation from the aqueous solutions of calcium nitrate and di-ammonium hydrogen phosphate salts. The precipitation process employed was also found to be suitable for the production of sub-micrometre HA/TCP composite powders in situ. The synthesized pure HA and TCP powders were found to be stable even at 1300 degrees C in air for prolonged heating times. Bioceramic sample characterization was achieved by powder X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and density and surface area measurements. Crystallographic analyses of HA powders were performed by the Rietveld method on the powder XRD data. PMID- 15348777 TI - The dependence of osteoblastic response on variations in the chemical composition and physical properties of hydroxyapatite. AB - Two synthetic hydroxyapatite powders (A and B), supplied by different manufacturers, were physically and chemically characterized before being die pressed and sintered at 1250 degrees C. The powders were characterized using X ray diffraction (XRD), infrared spectroscopy (IRS), X-ray fluorescence, surface area analysis (BET), particle size analysis and scanning electron microscopy (SEM). The materials were then pressed and sintered to produce hydroxyapatite discs of similar densities and grain sizes for in vitro evaluation. The ceramics were seeded with osteoblastic cells and after 15 days in culture the cell morphology was assessed using scanning electron microscopy (SEM), the ultrastructure of the cells was studied using transmission electron microscopy (TEM) with EDAX, and the rate of cell growth was assessed using biochemical techniques. The results clearly showed that the rate of cell proliferation but not the rate of alkaline phosphatase production, was highly dependent on the composition of the hydroxyapatite powders that were used to make the ceramic discs. The ultrastructural studies confirmed the relative viabilities of the cells and the nature of the ceramic interface indicating visually the marked differences in the performance of the two materials. PMID- 15348778 TI - The response of primary rat and human osteoblasts and an immortalized rat osteoblast cell line to orthopaedic materials: comparative sensitivity of several toxicity indices. AB - When studying the biocompatibility of orthopaedic biomaterials it is often necessary to discriminate between responses which show mild cytotoxicity. It is therefore essential to use a very sensitive index of toxicity. We have compared the sensitivity of four well-established indices of toxicity: total cell protein content, leakage of lactate dehydrogenase (LDH), reduced glutathione content and the MTT assay, with that of a novel index, alkaline phosphatase (ALP) activity. Comparisons were made by detecting nickel chloride toxicity in osteoblasts. ALP activity, the novel method, proved the most sensitive index of toxicity and it provides a convenient automated assay for assessing the interactions of materials with osteoblasts. The responses to nickel chloride and to aqueous extracts prepared from carbon fibre reinforced epoxy and polyetheretherketone (peek), two candidate materials for orthopaedic implants, were compared in primary and immortalized rat osteoblasts, and in primary human osteoblasts. Although the immortalized rat osteoblast cell line, FFC, was consistently the most sensitive cell type, the responses of the human cells and the FFC cell line were similar in terms of ALP activity throughout the range of nickel concentrations studied. Neither peek nor epoxy material extracts showed a significant decrease in the MTT or ALP responses in any of the three cell types. Our data suggest that immortalized rat osteoblasts may provide an in vitro model system for screening the biocompatibility of orthopaedic polymers. PMID- 15348779 TI - New Y-TZP powders for medical grade zirconia. AB - There is interest in using zirconia for biomedical applications as ball heads for total hip prostheses. Two potential types are under discussion: partially stabilized zirconia (PSZ) and tetragonal zirconia polycrystals (TZP) materials. Because of its enhanced material properties, TZP stabilized with yttria is favourable. To eliminate high amounts of natural radioactive impurities, the precursors are purified. The kind of precursor and purification method determine the powder impurity level. The disadvantage of Y-TZP is that the hydrothermal decomposition reaction method is that it depends very strongly on the grain size and the distribution of the stabilizing yttria within the zirconia grains. Thermodynamical and kinetic investigations on high purity coprecipitated and yttria-coated zirconia powders show different behaviours. Y-TZP materials based on yttria-coated zirconia powders show excellent mechanical strength of more than 1000 MPa, a Weibull modulus of up to 20 a! nd a fracture toughness of 9 MPa radical m. The material properties of Y-TZP ceramics based on coprecipitated powders and prepared under the same conditions are less attractive. It is expected that materials based on yttria-coated zirconia will show enhanced properties compared to materials derived from coprecipitated powders. Therefore Y TZP materials derived from yttria-coated powders are very attractive as medical grade zirconia. PMID- 15348780 TI - Bone response to surface modified titanium implants - studies on the tissue response after 1 year to machined and electropolished implants with different oxide thicknesses. AB - The bone formation around titanium implants with varied surface properties was investigated after 1 year in rabbits. Machined and electropolished samples with and without thick, anodically formed surface oxides were prepared, surface characterized and inserted in the cortical bone of rabbits. Scanning electron microscopy, scanning Auger electron spectroscopy and atomic force microscopy revealed marked differences in oxide thickness, surface topography and roughness, but no significant differences in surface chemical composition between the different groups of implants. Light microscopic morphology and morphometry showed that all implants were in contact with bone and had a large proportion of bone within the threads. There were no significant differences between the differently prepared implant groups. Our study shows that a high degree of bone contact and bone formation is achieved after 1 year with titanium implants which are modified with respect to oxide thickness and surface topography. There is no indication that a reduction of surface roughness, which in the initial phase decreases the rate of bone formation, had any influence on the amount of bone after 1 year in rabbit cortical bone. PMID- 15348781 TI - Biomechanical assessment of bone ingrowth in porous hydroxyapatite. AB - Porous hydroxyapatite (Endobon) specimens were implanted into the femoral condyle of New Zealand White rabbits for up to 6 months. After sacrifice, specimens were sectioned for histology and mechanical testing, where the extent of reinforcement by bony ingrowth was assessed by compression testing and fixation was assessed by push-out testing. From histological observations, it was established that the majority of bone ingrowth occurred between 10 day and 5 weeks after implantation and proceeded predominantly from the deep end of the trephined defect, with some integration from the circumferential sides. At 3 months, the implants were fully integrated, exhibiting bony ingrowth, vascularization and bone marrow stroma within the internal macropores. After 5 weeks, the mean ultimate compressive strength of retrieved implants (6.9 MPa) was found to be greater than that of the original implant (2.2 MPa), and by 3 months the fully integrated implants attained a compressive strength of approximately 20 MPa. Push-out testing demonstrated that after 5 weeks in vivo, the interfacial shear strength reached 3.2 MPa, increasing to 7.3 MPa at 3 and 6 months. PMID- 15348782 TI - Monocyte-biomaterial interaction inducing phenotypic dynamics of monocytes: a possible role of monocyte subsets in biocompatibility. AB - For the in vitro study of cell-biomaterial surface interactions, the choice of cell type is crucial. In vivo data indicate that during the healing of the implant in the tissues, the pivotal cell types are the macrophages. These cells, upon interaction with any foreign material, might initiate a spectrum of responses, which could lead to acute and chronic inflammatory changes affecting the biocompatibility of the implant. Whether the mechanisms governing the type of evolving inflammatory reaction could be attributed to the macrophages functional differentiation mirrored by monocyte subsets during the polymer interaction, is poorly described. This in vitro study, therefore, attempted to investigate whether different biomaterials influence monocyte cellular activity, determined by the myeloperoxidase level and mitochondrial XTT cleavage, and phenotype dynamics characterized by the presence of CD14, RM 3/1 and 27E10 antigens. It is shown that different polymers exert differential potential to influence monocytes, both in their cellular activity and their phenotypic pattern. Thus, these findings demonstrating material-induced monocyte activation and monocyte phenotype modulation, are suggestive of the monocyte role as reporter cells in evaluating the biocompatibility of a synthetic medical device. PMID- 15348783 TI - Complexation of polymeric drugs based on polyacrylic chains with aminosalicylic acid side groups. AB - Polymeric drugs and ionomers can exhibit properties associated with the polymeric nature, such as adhesion, or coating of different substrates by polymeric active films, and can be applied successfully in the field of biomaterials. In this sense, complexation with calcium ions of two polyacrylic systems derived from 4 aminosalicylic acid (poly (4-HMA)), and 5-aminosalicylic acid, (poly (5-HMA)), have been studied by Fourier transform-infrared spectroscopy and thermogravimetric analysis. Both techniques support the ability of these polymers to form complexes with calcium ions as polycarboxylates. PMID- 15348784 TI - Activation status of platelet aggregates and platelet microparticles shed in sheared whole blood. AB - The role of temperature and shear rate in the activation status of aggregating platelets and platelet microparticles (MPs) was investigated in a modified concentric-cylinder rotational viscometer. Whole blood anticoagulated with citrate was exposed to a range of shear rates typical of cardiopulmonary bypass circuits (0, 1000, 2000 and 4000 s(-1)) over four temperatures spanning hypothermic to mildly hyperthermic conditions (24, 30, 37 and 42 degrees C) for short durations (100 s). Aliquots of blood were double-stained for CD41 (platelet GPIIb/IIIa) and CD62 (P-selectin). Platelets, platelet aggregates, MPs and red blood cell-platelet and -MP aggregates were identified by flow cytometry by acquiring only CD41-positive particles and differentiating on a plot of CD41 versus forward light scatter. The activation status of each particle was quantified by measuring CD62 expression (alpha-granule release). A degree of correlation between the shedding of MPs and the formation of platelet-platelet aggregates was observed for the data as a whole (r=0.85 for p<0.01), although this trend was not observed for a shear rate of 4000 s(-1). The mean expression of CD62 on both platelets and MPs was maintained at a very low level for all temperature and shear rate combinations. There was, however, a number of very highly activated MPs associated with red blood cells at high shear rates. PMID- 15348785 TI - A new composite made of polyurethane and glass ceramic in a loaded implant model: a biomechanical and histological analysis. AB - The biocompatibility and osseous integration of a new composite material made of polyurethane and a calcium silicophosphate ceramic was investigated in a loaded implant model in sheep and compared to that of commercially pure titanium. Six months after implantation, interfacial shear strength was higher between the titanium and bone than between the composite and bone. After 2 years, however, the shear strength was significantly higher in the composite group. Histologically, both implants were surrounded by bone and fibrous tissue and there were no signs of inflammation. Direct contact of bone on the composite surface increased significantly with time, whereas there was no time-dependent increase of bone contact on titanium. It can be concluded that the biocompatibility and osseous integration of the composite was very good in the loaded implant model used. It is therefore suggested that the new composite is a promising biomaterial for orthopaedic implants. PMID- 15348786 TI - Surface characterization and biological evaluation of spark-eroded surfaces. AB - Forty commercially pure titanium implants were prepared with a spark-eroding process in order to create highly increased surface roughnesses. Two degrees of roughness were achieved by altering the applied current. Surface topographical characterization was performed with SEM and an optical profilometer. The surface composition and oxide layer were investigated using Auger scanning microscopy. In the present study, there was a large difference between the stipulated and measured surface roughness, indicating the need for a careful surface characterization in each new study. After 12 wk in rabbit bone, no statistically significant difference was found with respect to peak removal torque and histomorphometric analyses. The results from the present study provide no support for further increase of the surface roughness than that possible to achieve with a blasting technique. PMID- 15348787 TI - A simple method to prepare calcium phosphate coatings on Ti6Al4V. AB - A two-step chemical treatment followed by immersion in a supersaturated calcification solution (SCS) was found to be a simple way to prepare calcium phosphate (Ca-P) coatings on Ti6Al4V. The Ca-P deposition on the treated metallic surfaces could be accelerated by employing a pre-calcification (Pre-Ca) procedure prior to immersion in SCS. The two-step treatment was performed by etching the metallic plates with a mixture of HCl and H2SO4 followed by ageing in boiling diluted NaOH solution at 140 degrees C. Pre-Ca was carried out by incubating the two-step treated plates in Na2HPO4 solution and then in saturated Ca(OH)2 solution. The formation of a bioactive microporous surface oxide layer on Ti6Al4V by the two-step treatment was most probably responsible for the induction of Ca-P precipitation. The deposition rates and compositions of Ca-P coatings in two different SCSs were investigated by means of scanning electron microscopy, X-ray diffraction and infrared spectrophotometry. PMID- 15348788 TI - In vitro complement activation after contact with pyrolytic carbon-coated and uncoated polyethylene terephthalate. AB - This study was undertaken to evaluate whether the pyrolytic carbon coating of polyethylene terephthalate induces complement activation. Complement activation induced by pyrolytic carbon-coated polyethylene terephthalate (PET+PC) in comparison with uncoated polyethylene terephthalate (PET) was assessed on whole blood collected with heparin. The activation of the classic pathway was evaluated by C4d fragment enzyme immunoassay. The activation of the alternative pathway was evaluated with Bb fragment enzyme immunoassay. The results show that uncoated PET activates the alternative pathway, but not the classic one. PET+PC does not induce complement activation, not even through the alternative pathway. Pyrolytic carbon coating therefore contributes to improving blood compatibility. PMID- 15348789 TI - In vitro mechanical and biological assessment of hydroxyapatite-reinforced polyethylene composite. AB - In vitro performance of hydroxyapatite (HA)-reinforced polyethylene (PE) composite (HAPEX) has been characterized from both mechanical and biological aspects. The mechanical properties of HAPEX, such as tensile strength and Young's modulus, showed little change after immersion in a physiological solution at 37 and 70 degrees C for various periods. In addition, the biological response of primary human osteoblast-like (HOB) cells in vitro on HAPEX was assessed by measuring DNA synthesis and osteoblast phenotype expression. Cell proliferation rate on HAPEX was demonstrated by an increase in DNA content with time. A high tritiated thymidine ([3H]-TdR) incorporation/DNA rate was observed on day 1 for HAPEX, indicating a stimulatory effect on cell proliferation. The alkaline phosphatase (ALP) activity was expressed earlier on HAPEX than on unfilled PE and increased with time, indicating that HOB cells had commenced differentiation. Furthermore, it was found that the HA particles in the composite provided favourable sites for cell attachment. It appears that the presence of HA particles in HAPEX may have the advantage of acting as microanchors for bone bonding in vivo. PMID- 15348790 TI - Characterization of microblasted and reactive ion etched surfaces on the commercially pure metals niobium, tantalum and titanium. AB - In surface-roughened metallic implant materials, the topography, chemistry and energy of the surfaces play an important role for the cell and tissue attachment. The highly reactive commercially pure metals niobium, tantalum and titanium were analysed after microblasting (with Al2O3 powder and consecutive shot-peening with ZrSiO2), and after additional reactive ion etching (RIE, with CF4). Scanning electron microscopy in combination with energy-dispersive X-ray analysis and surface roughness measurements showed, for all microblasted surfaces, a heterogeneous roughening (Ra about 0.7 microm), and a contamination with blasting particles. RIE resulted in a further roughening (Ra about 1.1 microm), and a total cleaning from contaminations, except for traces of aluminium. Determination of surface energy by dynamic contact angle measurements showed an increase in surface energy after microblasting, which further increased after RIE, most pronounced for commercially pure niobium. In conjunction with superior electrochemical properties, this makes niobium and tantalum promising candidates for implant purposes, at least equal to the generally used titanium. PMID- 15348791 TI - A biomechanical regulatory model for periprosthetic fibrous-tissue differentiation. AB - Loosening of implants in bone is commonly associated with a development of fibrous interface tissues, due to interface gaps and a lack of mechanical stability. It has been postulated that the differentiation of these tissues to fibrocartilage or bone is governed by mechanical stimuli. The objective of our research is to unravel these relationships to the extent that the question whether an implant will loosen can be answered from initial conditions determined by implant and interface morphology, and functional loads. In this project we studied the hypothesis that distortional strain and interstitial fluid flow are the mechanical stimuli governing tissue differentiation. For that purpose, a biomechanical regulatory model was developed and used in association with a finite element code to simulate animal experiments with implants moving in bone. The similarities between the implant incorporation process found in the experiment and its simulation with the computer model demonstrate that our hypothesis is viable from a regulatory point of view. PMID- 15348792 TI - Immobilization of human thrombomodulin onto PTFE. AB - Human thrombomodulin (hTM) is an endothelial cell-surface glycoprotein and has effective anticoagulant properties. This protein was immobilized onto polytetrafluorethylene (PTFE) surfaces to create biomaterials with enhanced haemocompatibility. The PTFE surface was functionalized by CO2 plasma activation and subsequent vapour-phase graft polymerization of acrylic acid. Surface characterization after plasma treatment, grafting and hTM immobilization was achieved by attenuated total reflection-Fourier transform-infrared spectroscopy, X-ray photoelectron spectroscopy, zeta potential and wetting measurements. The activity of immobilized hTM was estimated using the protein C activation test. PMID- 15348793 TI - Corrosion behaviour and mechanical properties of functionally gradient materials developed for possible hard-tissue applications. AB - Artificial hip joints have an average lifetime of 10 years due to aseptic loosening of the femoral stem attributed to polymeric wear debris; however, there is a steadily increasing demand from younger osteoarthritis patients aged between 15 and 40 year for a longer lasting joint of 25 years or more. Compliant layers incorporated into the acetabular cup generate elastohydrodynamic lubrication conditions between the bearing surfaces, reduce joint friction coefficients and wear debris production and could increase the average life of total hip replacements, and other human load-bearing joint replacements, i.e. total knee replacements. Poor adhesion between a fully dense substrate and the compliant layer has so far prevented any further exploitation. This work investigated the possibility of producing porous metallic, functionally gradient type acetabular cups using powder metallurgy techniques - where a porous surface was supported by a denser core - into which the compliant layers could be incorporated. The corrosion behaviour and mechanical properties of three biomedically approved alloys containing two levels of total porosity (>30% and <10%) were established, resulting in Ti-6Al-4V being identified as the most promising biocompatible functionally graded material, not only for this application but for other hard tissue implants. PMID- 15348794 TI - Release of tobramycin from tobramycin-containing bone cement in bone and serum of rabbits. AB - Tobramycin release from tobramycin-containing bone cement was studied in vivo in a rabbit model. After insertion of cement into the right femur, tobramycin concentration as a function of time for up to 28 days was measured in serum and bone of rabbits. Tobramycin release in the femoral cortex adjacent to the cement was found to exceed the minimal inhibitory concentration for Staphylococcus aureus Wood 46. Serum tobramycin concentrations were below the systemic toxicity threshold. PMID- 15348795 TI - Effect of crystal seeding on the hydration of calcium phosphate cement. AB - In this paper, the effect of crystal seeding on the hydration of calcium phosphate cement (CPC) has been carefully investigated. The setting time of the CPC slurry not containing any crystal seeds was 150 min, while the setting time for the specimen containing 5 wt% low crystallinity hydroxyapatite used as a crystal seed was 7 min. This improvement in the setting time was due to HAP serving as a substrate for heterogeneous nucleation which accelerated nucleation. In addition, the compressive strength of the specimen containing the crystal seeding was deduced and we report values different from those previously reported in the literature. The calorimetric curve indicated that crystal seeding could reduce the induction period. A.c. impedance spectroscopy revealed that at the beginning of hydration, the rate of reaction increased and also that the mean diameter and porosity decreased as the seed content increased. At the end of the hydration reaction the situation was changed with the mean diameter and porosity in the sample without any seeds being a minimum, which indicated that the compressive strength was a maximum. This result could be explained by the dissolution and reprecipitation of small hydration products produced by the high rate of reaction produced by the introduction of the crystal seeds. PMID- 15348796 TI - Evaluation of in vitro bioactivity and biocompatibility of Bioglass-reinforced polyethylene composite. AB - The bioactivity and biocompatibility of Bioglass-reinforced high-density polyethylene composite (Bioglass/HDPE) have been evaluated in simulated body fluid (SBF) and by in vitro cell culture, respectively. The formation of a biologically active hydroxy-carbonate apatite (HCA) layer on the composite surface after immersion in SBF was demonstrated by thin-film X-ray diffraction, infrared spectroscopy and scanning electron microscopy, indicating the in vitro bioactivity of Bioglass/HDPE composites. The HCA layer was formed on the 40 vol% composite surface within 3 days immersion in SBF at a formation rate comparable to those on bioactive glass-ceramics, showing that in vitro bioactivity could be obtained in a composite. Furthermore, the composite was biocompatible to primary human osteoblast-like cells. In comparison with unfilled HDPE and tissue culture plastic control, a significant increase in cellular metabolic activity was found on the composite. Therefore, Bioglass/HDPE composites have a promising biological response as a potential implant material. PMID- 15348797 TI - Guided bone regeneration using osteopatite granules and polytetrafluoroethylene membranes. AB - Granules of a modified hydroxyapatite, Osteopatite, were implanted in the right posterior tibiae of adult rabbits. We studied the extent of bone regeneration in bone holes. In the right tibiae, that were filled with granules of this biomaterial covered with a polytetrafluoro-ethylene (PTFE) membrane using, as a control, uncovered granules. In the left tibia, an empty hole was covered with PTFE membrane and a second hole was left empty to be used as a control. A histomorphometric study was carried out using light microscopy, four and eight weeks after the surgery. The covered granules presented a higher percentage of bone contact than the uncovered ones, and it was also possible to observe a better bone tissue organization, mainly produced by the immobilization action of the PTFE membrane. Empty bone defects covered with PTFE membranes, two months after implantation, presented large areas of Haversian bone and direct bone contact to the PTFE membrane. PMID- 15348798 TI - Ex vivo expansion of haematopoietic progenitors on an endothelialized hydroxyapatite matrix. AB - Autologous haematopoietic progenitor cell (HPC) transplantation is increasingly used to restore haematopoiesis after high-dose chemotherapy treatments. The present study was designed to analyse the ability of hydroxyapatite (HAP) seeded with endothelial cells (EC) to support the proliferation and differentiation of CD34+ HPC in static culture conditions. HAP is endothelializable as assessed by scanning electron microscopy and time-course DNA synthesis analysis using tritiated thymidine incorporated in EC isolated from human umbilical vein cord. Short-term coculture experiments in which CD34+ cells isolated from human cord blood were seeded on endothelialized HAP, were performed. Results show that endothelialized HAP is permissive to CD34+ cell expansion with a maximum expansion obtained between days 7 and 14 of coculture in the presence of IL-1 and IL-3 when compared with other experiments omitting either EC or interleukins. From morphological analyses, the expanded cell population mainly belonged to the myelocytic lineage with 33% mature cells (polymorphonuclear neutrophils and monocytes) at day 14 of coculture. The immature HPC could remain trapped within HAP while giving rise to a more mature progeny that exit from HAP microenvironment. PMID- 15348799 TI - Responses of bone to titania-hydroxyapatite composite and nacreous implants: a preliminary comparison by in situ hybridization. AB - The effect of two biomaterials on bone formation in vivo by in situ hybridization, was compared by using RNA probes complementary to collagen alpha1(I) RNA, osteonectin RNA and osteocalcin RNA. Holes were drilled into the midshafts of rat femurs. Titania-hydroxyapatite composite (THA) or nacre cylinders were implanted and the bone-implant regions collected 14 days after operation. Cuboidal osteoblasts, intensely labelled with the three probes, were seen to be lining the newly formed bone surrounding the THA implant. Between the implant and the new bone, a layer of un-labelled, apparently non-osteogenic cells was observed. By contrast, the nacre implant was bonded to the newly formed bone without any soft tissue interference. Osteoblasts lining the distal surface of the newly formed bone were stained with all three RNA probes, although weaker than in the THA sample. Some of the osteoblasts were flattened. We concluded from the appearance of the osteoblasts that the bone formation in the nacre samples had progressed beyond the phase of maximal synthetic activity. Around the THA implant, the labelling indicated that bone-forming activity was still high. It was concluded that the bioactivity of nacre was higher than that of THA. PMID- 15348800 TI - Effect of crosslinking agents on poly(ethylmethacrylate) bone cements. AB - Asceptic loosening of cemented joint prostheses in many cases is related to the mechanical failure of the acrylic bone cement. Poly(methylmethacrylate) bone cements are widely used in orthopaedic surgery although there are well-known disadvantages. A lower modulus bone cement based on poly(ethylmethacrylate)-n butylmethacrylate with a lower polymerization exotherm, and a low monomer extractibility, is a promising alternative. The effect of incorporating crosslinking agents in order to improve the mechanical performance of the PEMA bone cement is reported. Three different bifunctional dimethacrylate crosslinking agents with different chain lengths and degrees of flexibility were incorporated in the monomer phase, and cements formulated. The setting time was found to decrease in the presence of the cross-linking agents and the polymerization exotherm decreased in the presence of triethylene glycol dimethacrylate and polyethylene glycol dimethacrylate, n=400. Incorporation of triethylene glycol dimethacrylate showed an increase in the tensile strength and modulus with a decrease in the strain at maximum stress. However, polyethylene glycol dimethacrylate, n=400, did not improve the mechanical properties appreciably which may be attributed to the low crosslinking density and higher flexibility of the spacer group in the crosslinking agent. PMID- 15348801 TI - From cytotoxicity to biocompatibility testing in vitro: cell adhesion molecule expression defines a new set of parameters. AB - Determination of potential cytotoxicity is a central issue in current biocompatibility testing standards such as ISO and ASTM. Most of these tests do not assess biocompatibility of a biomaterial with regard to cell function. This study was aimed at screening a number of potential parameters that could be included in assessment of cell functional aspects of biocompatibility. Human umbilical vein endothelial cells (HUVEC) were seeded directly on titanium, NiCr alloy, CoCr alloy, PMMA, PE, PU, PVC, and silicone, or were exposed to the material extracts. Cytotoxicity was assessed for these materials through MTT conversion, crystal violet protein determination and Ki67 expression. In addition, expression of the cell adhesion molecules E-selectin, cadherin-5 and PECAM, as well as of the adhesion-associated proteins fibronectin and vinculin (focal adhesions), was determined by immunocytochemistry and western blotting. Cytotoxicity was not detected with the material extracts. Cells were able to adhere to bare metals, but not polymers. Fibronectin preadsorption resulted in adhesion and spreading also on the polymers. Cells were able to establish cell cell contacts and focal adhesions. Western blotting, in combination with differential detergent extraction, indicated that linkage of cell-cell adhesion markers to the cytoskeleton may be used as an additional parameter relevant to cell function. PMID- 15348802 TI - Failure patterns of four osseointegrated oral implant systems. AB - The aim of this metanalysis was to investigate possible differences in failure patterns among four different osseointegrated oral implant systems. Only systems with a supposed scientific validation, based on long-term follow-up studies, were selected for this analysis, i.e. the Branemark system, the Calcitek Integral system, the Interpore IMZ system and the Straumann ITI system. While several prospective reports could be found on the Branemark system, only a few retrospective investigations concerning the ITI and the IMZ systems were available. No data on the Integral system could be employed. Despite these limitations, a substantial difference in failure patterns among various implant systems was observed. The Branemark implants showed a higher incidence of early failures, though sharply decreasing over time. On the contrary, IMZ implant characterized by rougher surfaces displayed a lower incidence of early failures, but showed constant or increased failure rates over time. For the ITI implants, for example, a higher prevalence of late failures, attributable to chronic bacterial infection (peri-implantitis), was observed when compared to the Branemark system. With the exception of the latter implant system, prospective long-term follow-up studies, using similar and well-defined success criteria, are needed for the others to confirm the current preliminary findings. PMID- 15348803 TI - Influence of sterilization upon a range of properties of experimental bone cements. AB - Bone cement, used to fix prostheses into the bone, must be sterilized prior to implantation. Two sterilization techniques, gamma and beta radiation, were investigated, examining the influence upon molecular weight, static and dynamic mechanical characteristics and rheological properties. A number of experimental cements were studied prepared from methylmethacrylate (MMA) co-polymers, either single powders or powder blends, mixed with MMA monomer. It was found that with both gamma and beta radiation, there was a decrease in molecular weight of all powders, including a MMA/styrene co-polymer, in relation to the radiation dose. This fall in molecular weight resulted in a drop in tensile strength, Young's modulus and strain to failure of all cements tested. However, the deterioration in mechanical strength was highlighted by the dynamic testing. Fatigue lives of cements after testing in tension-tension, at 2 Hz under load control and irradiated with 25 kGy gamma radiation, displayed significant decreases. This result indicated the utmost importance of conducting such tests upon experimental bone cements prior to in vivo use. The rheological time profiles of curing cements were also found to be influenced by 25 kGy gamma radiation, with a reduction of complex viscosity after sterilization. PMID- 15348804 TI - Acutely repaired proximal anterior cruciate ligament ruptures in sheep - by augmentation improved stability and reduction of cartilage damage. AB - The aim of this study was to evaluate the healing capacity of proximal anterior cruciate ligament (ACL) ruptures following primary repair with and without a bioresorbable augmentation. The ACL was transected at the femoral origin in the right knee joint of 24 sheep. The ACL was repaired in eight sheep (group B) without, and in eight sheep (group C) with a bioresorbable augmentation. Eight sheep without repair served as a control (group A). No immobilization was performed in any group. The sheep were sacrificed 13 weeks post-operatively. Macroscopically, all repaired ACLs were healed. The augmentation device was broken in all cases, but not completely degraded. In group A, none of the transected ACLs had healed. The anterior drawer under a load of 50 N was significantly lower in group C than in group A (p<0.01). No significant difference was seen between groups B and A. The distribution and extent of chondromalacia (CM) in the operated knees depended on the type of operative treatment (p<0.01). Groups A and B showed significantly more CM in the operated knee than in the non-operated knee (each p<0.05). Proximal ACL ruptures can heal in sheep after both non-augmented and augmented ACL repair with free-functional rehabilitation. However, augmented repair leads to significantly better stability of the knee joint compared to transected controls and better limits the development of degenerative changes. PMID- 15348805 TI - Hydroxyapatite coatings: a comparative study between plasma-spray and pulsed laser deposition techniques. AB - A comparative study between hydroxyapatite coatings produced by two different techniques, plasma spray (PS) and pulsed-laser deposition (PLD) was carried out. Plasma spray is currently commercially used for coating dental and orthopaedical implant devices, and pulsed-laser deposition (or laser-ablation deposition) gave good results in the field of high critical temperature superconductive thin films, and is being applied to produce calcium phosphate coatings for biomedical purposes. X-ray diffraction was used to control the crystallinity of the coatings, scanning electron microscopy for the surface and cross-sectional morphology, and the pull test to determine the tensile strength of the coatings. This study reveals that the pulsed-laser deposition technique appears to be a very good candidate to replace the plasma spray in many biomedical applications, because it overcomes most of the drawbacks of the plasma spray. PMID- 15348806 TI - Immobilization of the cell-adhesive peptide Arg-Gly-Asp-Cys (RGDC) on titanium surfaces by covalent chemical attachment. AB - Surface modification of acid-pretreated titanium with 3 aminopropyltriethoxylsilane (APTES) in dry toluene resulted in covalently bonded siloxane films with surface coverage that was relatively controllable by regulating the reaction conditions. A hetero-bifunctional cross-linker, N succinimidyl-3-maleimidopropionate (SMP), reacted with the terminal amino groups, forming the exposed maleimide groups. Finally, a model cell-binding peptide, Arg Gly-Asp-Cys (RGDC), was immobilized on the surface through covalent addition of the cysteine thiol groups to the maleimide groups. X-ray photoelectron spectroscopy, radiolabelling techniques, and ellipsometry were used to quantify and characterize the modified surfaces. PMID- 15348807 TI - Structural analysis of calcium phosphate coatings produced by pulsed laser deposition at different water-vapour pressures. AB - Calcium phosphate coatings have been produced by pulsed laser deposition (PLD) at different water-vapour pressures. Rietveld refinement of X-ray diffraction (XRD) data allows us to determine that the structure of these coatings is apatitic with carbonate substitution for phosphate. The carbonate substitution decreases when the chamber pressure is raised, a fact that has been corroborated by Fourier transform-infrared (FT-IR) spectroscopy. Carbonate concentrations between 5 and 17 wt% have been calculated for the crystalline samples. Amorphous coatings are produced at the lowest and highest pressures due to the high carbonate concentration in the first case, and possibly to another type of substitution (Mg(2+), HPO(2-)4, P2O(4-)7) or the inherent kinetics of the PLD process, in the second case. PMID- 15348808 TI - Up-regulation of thrombomodulin is inducible on an endothelialized polyester. AB - The concept of endothelial cell seeding of vascular prostheses is designed as a method for improving long-term patency substitutes because endothelium is considered as the haemocompatible surface of reference. The assessment of the functionality of cells lining a biomaterial is thus of crucial importance. We have reported encouraging results concerning the ability of a polyester coated with albumin and chitosan (M 11) to be lined by a confluent monolayer of cultured human endothelial cells (EC). The aim of the present work was to study the expression of thrombomodulin (membrane glycoprotein responsible for anticoagulant activity) in EC lying on M.11 by anticoagulant activity and mRNA level with and without stimulation. Results obtained in basal conditions showed that EC on M.11 have a comparable expression of TM mRNA when compared to control (EC on tissue culture plates) despite a lower TM surface activity for EC on the biomaterial. In terms of ratio (stimulated cells to unstimulated ones) the response in activity for EC on M.11 is comparable to that of the control. These results indicate that cells lying on M.11 are able to respond to physiological-like stimuli, despite a tendency for these cells to express a procoagulant phenotype when compared with control EC. PMID- 15348809 TI - NaOH treatment of vacuum-plasma-sprayed titanium on carbon fibre-reinforced poly(etheretherketone). AB - Carbon fibre-reinforced polyetheretherketone (CF-PEEK) substrates were coated with titanium by vacuum-plasma-spraying and chemically treated in 10 M sodium hydroxide (NaOH) solution. After NaOH treatment, the specimens were immersed in simulated body fluid (SBF) containing ions in concentrations similar to those of human blood plasma. Scanning electron microscopy, energy-dispersive X-ray analysis and diffuse reflectance Fourier transformed-infrared spectroscopy were used to analyse the NaOH-treated VPS-Ti surface and the calcium phosphate layer formed during immersion in SBF. It was observed that a carbonate-containing calcium phosphate layer was formed on the NaOH-treated VPS-Ti surface during immersion in SBF, whereas no calcium phosphate precipitation occurred on the untreated surfaces. It is therefore concluded that vacuum-plasma-spraying with titanium and subsequent chemical modification in 10 M NaOH solution at 60 degrees C for 2 h is a suitable method for the preparation of bioactive coatings for bone ongrowth on CF-PEEK. PMID- 15348810 TI - The effect of temperature and shear rate on platelet aggregation. AB - Samples of whole blood were obtained from male volunteers and exposed to combinations of shear rates and temperatures representative of cardiopulmonary bypass (CPB) in a modified computer-controlled concentric cylinder rotational viscometer for a period of 100 s. Blood sampled from the chamber was fixed in paraformaldehyde, stained with CD41 and analysed by flow cytometry. Only platelet positive particles were acquired, each individual cell, or aggregate of cells, identified by analysis of its fluorescence and forward light scatter characteristics. Little platelet aggregation was observed at shear rates of less than 4000 s(-1) for temperatures of greater than 24 degrees C, but large numbers of aggregates were formed at all temperatures at 4000 s(-1) (p<0.05), with more aggregates forming at 24 and 30 degrees C than at 37 and 42 degrees C (p<0.05). We conclude that the process of aggregation is dependent on both temperature and shear rate. We note that a large number of platelets become involved in aggregates under conditions of temperature and shear-rate typical of CPB. PMID- 15348811 TI - Topographical characterization and microstructural interface analysis of vacuum plasma-sprayed titanium and hydroxyapatite coatings on carbon fibre-reinforced poly(etheretherketone). AB - In the present study, topographical characterization and microstructural interface analysis of vacuum-plasma-sprayed titanium and hydroxyapatite (HA) coatings on carbon fibre-reinforced polyetheretherketone (CF/PEEK) was performed. VPS-Ti coatings with high roughness values (Ra=28.29+/-3.07 microm, Rz=145.35+/ 9.88 microm) were obtained. On this titanium, intermediate layer HA coatings of various thicknesses were produced. With increasing coating thickness, roughness values of the HA coatings decreased. A high increase of profile length ratio, Lr, of the VPS-Ti coatings (Lr=1.45) compared to the grit-blasted CF/PEEK substrate (Lr=1.08) was observed. Increasing the HA coating thickness resulted in a reduction of the Lr values similar to the roughness values. Fractal analysis of the obtained roughness profiles revealed that the VPS-Ti coatings showed the highest fractal dimension of D=1.34+/-0.02. Fractal dimension dropped to a value of 1.23-1.25 for all HA coatings. No physical deterioration of the CF/PEEK substrate was observed, indicating that substrate drying and the used VPS process parameter led to the desired coatings on the composite material. Cross-section analysis revealed a good interlocking between the titanium intermediate layer and the PEEK substrate. It is therefore assumed that this interlocking results in suitable mechanical adhesive strength. From the results obtained in this study it is concluded that VPS is a suitable method for manufacturing HA coatings on carbon fibre-reinforced PEEK implant materials. PMID- 15348812 TI - Treatments to induce the nucleation and growth of apatite-like layers on polymeric surfaces and foams. AB - In this work, a bioactive glass is used as a percusor of calcium-phosphate (Ca-P) film deposition onto several polymer-based materials. Both bioinert (high molecular weight polyethylene, HMWPE), and biodegradable (corn starch-based blends, SEVA-C) polymers, unreinforced or reinforced with hydroxylapatite (HA), were coated by the very simple proposed route. Also polyurethane (PU) foams, with an open-cell structure, were mineralized by the proposed method. In fact, it was possible to induce the growth of the Ca-P films not only at the surface, but also in the bulk of the PU foam. These cellular materials are intended for cancellous bone replacement applications. The morphology of the formed films was strongly dependent on the used substrate, its polar character, and on the presence of HA in its composition, as observed by SEM. Nevertheless, a well defined needly like structure was observed in all samples at high magnifications. The Ca:P ratios of the films were between 1.5 and 1.7, i.e. in the range of tricalcium phosphate hydroxylapatite. Raman spectroscopy and thin-film x-ray diffraction (XRD) evidenced the formation of mostly amorphous calcium-phosphate films. After scraping the coating from the polymer surface and heat-treating the resulting powder at 1000 degrees C for 1 h, HA and beta-tricalcium phosphate (TCP) typical peaks were found on XRD patterns. PMID- 15348813 TI - Hydroxyapatite-zirconia composites prepared by precipitation method. AB - Chemical routes to prepare ceramic precursor powders are frequently envisaged, especially when the aspects related to the microstructure are important and need to be controlled. An understanding of which parameters of synthesis and thermal treatment can influence the mechanical properties of hydroxyapatite compounds is essential for the production of such materials. Hydroxyapatite-zirconia composites have been prepared, in this study, by a precipitation method. This led to the formation of homogeneous powders with a very defined particle-size distribution. Ceramic pellets prepared from these powders were easily compacted and sintered without cracking. As expected, the presence of the zirconia phase improved composite densities and appeared to have an important role in thermal stabilization of the hydroxyapatite phase. PMID- 15348814 TI - Modulation of the phenotypic and functional properties of phagocytic macrophages by wear particles from orthopaedic implants. AB - An attempt was made to assess the local chronic inflammatory response in patients with failed orthopaedic implant that is clinically associated with osteolysis, bone and bone marrow necrosis. The main objective was to analyse the heterogeneity of the macrophage functional subsets in the bone-implant interface membrane and to evaluate their possible role in the development of an erosive inflammatory lesion within the bone. Immunohistology was performed on 21 specimens of the bone-implant interface obtained from 17 patients during revision arthroplasty, and synovial membranes from rheumatoid (RA, n=4), and osteoarthritis (OA, n=4) patients. Three well-characterized monoclonal antibodies (MAb) recognizing antigenic determinants on specific functional subsets of macrophages (Mphi) were used. RFD1 (interdigitating reticulum cells/antigen presenting cells, (APC), RFD7 (mature phagocytic macrophages), and RFD9 epithelioid cells and foreign body giant cells (FBGC). RFD1 was expressed on a variable number of perivascular and synovial lining Mphi in both RA and OA synovia, at a frequency of 25%-40%. In cases with total joint replacements, the interface showed a marked increase in the expression of RFD1 (20%-90%). A considerably greater percentage of RFD1 positive Mphi and FBGC was noted in the interfaces from cases with a high level of detectable metal particulate wear debris (mean 80%, range 60%-90%) than in cases with polyethylene wear debris (mean 30%, range 0%-50%), p 0.0001. RFD7 labelled most tissue Mphi in each group. Immunoreactivity for RFD9 was restricted to FBGC in all cases analysed. The finding of elevated expression of RFD1 on metal-containing Mphi and FBGC in the bone-implant interface suggests an increase in antigen-presenting phenotype and indicates that metal particles have more impact in the induction of immune mediated responses. Such responses are characterized by sustained cellular hyperreactivity and phenotypic changes in Mphi subsets. PMID- 15348815 TI - Quantitative rates of in vivo bone generation for Bioglass and hydroxyapatite particles as bone graft substitute. AB - Rates of in vivo bone generation were characterized by point-counting analysis of particulate Bioglass and synthetic hydroxyapatite (HA) in rabbit femora. New bony tissue was observed in approximately 20% of the image area around Bioglass particles at 1 wk, and the degree of trabecular bone growth increased with time. The interparticle space of Bioglass was filled by 80% bonding bone between 6 and 12 wk. The rate constants of trabecular bone growth in the presence of Bioglass were approximately 10.9x10(-3) d(-1) at the periphery of the implantation site. HA particles led to smaller rate constants of approximately 4.6x10(-3) d(-1) at the periphery, and the HA particles developed very small amounts of bridging bone. The quantitative rate of bone growth matched well with previously measured bioactive indices of the materials. PMID- 15348816 TI - Structure of the interface between rabbit cortical bone and implants of gold, zirconium and titanium. AB - The role of surface properties (chemical and structural) for the interaction between biomaterials and tissue is not yet understood. In the present study, implants made of titanium, zirconium (transition metals with surface oxides) and gold (metallic surface) were inserted into the rabbit tibia. Light microscopic (LM) morphometry showed that after 1 and 6 mo the gold implants had less amount of bone within the threads and a lower degree of bone-implant contact than the titanium and zirconium implants, which did not differ from each other. These quantitative differences were supported by LM and ultrastructural observations of the interface. The ultrastructural observations in addition demonstrated that the layer of non-collagenous amorphous material located between the implant and the calcified bone was appreciably thicker around zirconium than around titanium implants. The factors potentially responsible for the observed morphological differences in the bone around the different material surfaces are discussed. PMID- 15348817 TI - Heterocyclic methacrylates for clinical applications-further studies of water sorption. AB - The room temperature polymerizing system comprising poly(ethyl methacrylate) tetra hydrofurfuryl methacrylate (PEM/THFMA) has potential in orthopaedic and dental applications, and earlier work has shown it to have unusual water absorption characteristics. This aspect has been studied in further detail, by studying the water absorption behaviour from some biological solutions, and the effect of the addition of an antibiotic (gentamicin). For comparison purposes, a parallel system whereby tetrahydrofuryl methacrylate was replaced by hydroxyethyl methacrylate (PEM/HEMA), was studied. In the case of PEM/THFMA, water uptake was substantially reduced when absorption was carried out from solutions (from about 30% in water to about 1.5% in solutions of higher concentrations), and the corresponding diffusion coefficient increased (by a factor of several hundred). The addition of gentamicin increased uptake, but the extent of increase also decreased in solutions. It was concluded that uptake was related to the osmolarity of the external solution, and also on the presence of osmotic sites within the polymer; hence the uptake process appears to be governed by chemical potential considerations. At the higher uptakes, there was evidence of water clusters. In marked contrast, the uptake by the PEM/HEMA system was independent of the osmolarity of the external solutions, presumably due to the hydrophilic nature of HEMA. PMID- 15348818 TI - Composition effects on the pH of a hydraulic calcium phosphate cement. AB - The pH of a hydraulic calcium phosphate cement (HCPC) made of monocalcium phosphate monohydrate (Ca(H2PO4)2.H2O; MCPM), beta-tricalcium phosphate (beta (Ca3(PO4)2; beta-TCP) and water was measured as a function of reaction time and composition at room temperature. During setting, the cement pH varies from very acidic pH values, i.e., 2.5, to almost neutral pH values, i.e., 6. The cement pH profile significantly depends on the initial cement composition. However, all profiles are characterized by a sharp initial decrease of the pH due to the dissolution of MCPM crystals and the precipitation of dicalcium phosphate dihydrate (CaHPO4. 2H2O; DCPD) crystals. With an excess of MCPM, the final pH stays low, and its value can be predicted from the initial composition of the cement and solubility data. With an excess of beta-TCP, the end pH is close to 5, which is much lower than 5.9, the value predicted by calculation. Results suggest that the difference may be due to the presence of impurities in the cement. Replacing MCPM by phosphoric acid renders the cement paste very acidic for the initial 30 s, but then the pH profile follows that obtained with MCPM. Adding pyrophosphate ions into the cement paste postpones the position of the pH minimum. The delay, which is proportional to the concentration of pyrophosphate ions, is thought to be due to the inhibiting action of pyrophosphate ions on the precipitation of DCPD crystals. PMID- 15348819 TI - Surface activation of polyetheretherketone (PEEK) and formation of calcium phosphate coatings by precipitation. AB - Plasma activation of polyetheretherketone (PEEK) surfaces and the influence on coating formation in a supersaturated calcium phosphate solution was investigated in this study. It was observed that plasma treatment in a N2/O2 plasma had a significant effect on the wettability of the PEEK surface. The contact angle decreased from 85 degrees to 25 degrees after plasma treatment. Cell culture testing with osteoblastic cell lines showed plasma activation not to be disadvantageous to cell viability. X-ray photoelectron spectroscopy (XPS) analysis was performed to characterize the chemical composition of the PEEK surfaces. It was observed that the O1s intensity increased with plasma activation time. At the C1s peak the appearance of a shoulder at higher binding energies was observed. Coating of PEEK was performed in a supersaturated calcium phosphate solution. Coating thicknesses of up to 50 microm were achieved after 24 days of immersion. Plasma activation followed by nucleation in a highly saturated hydroxyapatite solution had a positive effect on the growth rate of the layer on PEEK. Chemical analysis revealed that the coating consists of a carbonate containing calcium phosphate. PMID- 15348820 TI - The physics of water sorption by resin-modified glass-ionomer dental cements. AB - The water-sorption characteristics of two commercial resin-modified glass-ionomer dental cements (Baseline VLC, ex. Detrey Dentsply, and Vitremer lining cement, ex. 3M Dental Products) have been studied in more detail than previously. Water sorption in both cements proved to be rapid, reaching equilibrium at approximately 48 h for Baseline VLC and at approximately 10 d for Vitremer. Over the first 8 h or so, absorption was shown to follow Fick's law, with a diffusion coefficient of 1.56x10(-7) cm2 s(-1) for Baseline VLC (cured for 20 s) and 5.09x10(-7) cm2 s(-1) for Vitremer (also cured for 20 s). As expected, sorption of water was found to be faster in specimens cured for shorter cure times and slower for those cured for longer times. In the presence of sodium chloride, both at 0.9% and at 1 M, diffusion coefficients were significantly greater than in pure water, but did not vary significantly with sodium chloride concentration, being approximately 3.3x10(-7) cm2 s(-)1 for Baseline VLC and 8.0x10(-7) cm2 s( 1) for Vitremer. This is attributed to conformational changes in hydrophilic segments of the polymer on absorption of aqueous sodium chloride in which the molecules form more compact coils than in the presence of pure water. They thus create a microstructure that is more permeable to water. Sorption in salt solutions became non-Fickian much sooner than in pure water, i.e. at 3-4 h for both cements. This is probably due to concentration changes of salt within the cement, suggesting that these materials possess a degree of permselectivity. Finally, equilibrium water uptakes varied with salt concentration, being least in 1 M NaCl, which reflects the different chemical potentials of water in the various storage media. PMID- 15348821 TI - Osteogenic responses to extraskeletally implanted synthetic porous calcium phosphate ceramics: an early stage histomorphological study in dogs. AB - In this experiment, synthetic porous calcium phosphate ceramics (hydroxyapatite tricalcium phosphate) were prepared and implanted in dorsal muscles of dogs. The purpose was to study the biological processes prior to and during the morphogenesis of bone in extraskeletally implanted porous calcium phosphate ceramics. Specimens were harvested after implantation for 7, 15, 30, 45, 60, 90 and 120 days. Decalcified and undecalcified sections were prepared for alkaline phosphatase (ALP) histochemical localization and comparative histological analysis. The results show that bone morphogenesis in the pore regions of the extraskeletally implanted ceramics follows a complex process involving clot formation, vascular invasion, granulation-like tissue formation, polymorphic cell aggregation, osteoblast differentiation and bone formation. The characteristic feature preceding bone formation was polymorphic cell aggregation on the pore inner surface and near the invading capillaries or small venules. These cells were of various sizes and shapes, and some of them were positive for ALP activity. ALP-positive cell aggregates were more numerous where capillaries or venules were close to the pore inner surface. Osteoblast differentiation occurred within the cell clusters aggregated on the pore inner surface and bone matrix was secreted in direct contact with the ceramics. During bone formation, capillaries or small venules were always found close to the developing fronts of the osseous nidi. It is suggested that those cells which first appeared near the invading vasculature, the cells which aggregated on the pore inner surface and those cells which finally differentiated into osteoblasts may be interrelated in some way. PMID- 15348822 TI - Effect of diluent upon the properties of a visible-light-cured dental composite. AB - Four long-chain aliphatic organic diluents, triethylene glycol dimethacrylate (TEGDMA), ethylene glycol dimethacrylate (EDMA), triethyleneglycol diacrylate (TEGDA) and tetraethylene glycol diacrylate (TTEGDA) were used as diluents for preparing a visible-light-cured dental composite. Cured samples of pastes prepared using the above diluents were stored separately for a period of 60 days in distilled water at 37 degrees C and tested for changes in compressive strength, diametral tensile strength and microhardness at specific intervals of 1, 7, 14, 21, 28 and 60 days. Samples prepared using TEGDMA showed better strength values initially compared with the composite specimens prepared using the rest of the diluents although they are found to deteriorate slowly upon storage in distilled water up to 60 days. Samples prepared using EDMA showed retention in properties whereas samples prepared using TEGDA and TTEGDA showed large deviations in strength values during the first 28 days, did not deteriorate further and were found either to show improvement upon ageing or to remain constant. Sorption values seem to reach a maximum for all composites and then to decrease further after 14 days whereas solubility is found to increase with time. PMID- 15348823 TI - A novel method for fabrication of biodegradable scaffolds with high compression moduli. AB - It has been previously shown that, when used for meniscal reconstruction, porous copoly(L-lactide/epsilon-caprolactone) implants enhanced healing of meniscal lesions owing to their excellent adhesive properties. However, it appeared that the materials had an insufficient compression modulus to accomplish 100% fibrocartilage formation. In addition, to be used for meniscal prosthesis, the compression modulus of the porous materials should be larger than 150 kPa in order to protect the articular cartilage. A technique was developed to prepare stiff porous materials of a high molecular weight 50/50 copoly(L-lactide/epsilon caprolactone) suitable for fibrocartilage regeneration in meniscal implants and meniscal prosthesis. Porous microspheres (50-250 microm) were agglutinated in the presence of NaCl crystals (250-300 microm). The microspheres were mixed with solid solvent in order to obtain a homogeneous distribution of solvent over the spheres. By changing the amount of solvent and crystals, the density and the compression modulus could be varied over a range of 0.07 g ml(-1) to 0.5 g dl(-1) and 40-1100 kPa, respectively. PMID- 15348824 TI - Sequential polyurethane-poly(methylmethacrylate) interpenetrating polymer networks as ureteral biomaterials: mechanical properties and comparative resistance to urinary encrustation. AB - The mechanical properties and resistance to urinary encrustation of sequential interpenetrating polymer networks (IPNs) composed of polyurethane (PU) and polymethylmethacrylate (PMMA), have been described. Mechanical properties were determined using tensile testing and dynamic mechanical analysis, whereas resistance to encrustation was examined using an in vitro model for encrustation simulating in vivo encrustation. Maximum and minimum tensile strength at break, Young's modulus, storage and loss moduli were associated with PMMA and PU, respectively. IPNs demonstrated intermediate mechanical properties which were dependent on the concentrations of the component polymers. Conversely, maximum elongation at break was observed for PU and this parameter decreased as the concentration of PMMA increased in the IPN. The dynamic mechanical damping parameter, tan delta, was similar for all IPNs at 37 degrees C. Increased advancing and decreased receding contact angles were observed for IPNs in comparison with the native PU. The rate and extent of encrustation, measured as the percentage surface coverage, was similar for PU, IPNs and PMMA. In contrast, encrustation on polyhydroxyethylmethacrylate, a model hydrogel, was greater than observed for the IPNs or component polymers. No apparent correlation was observed between the rate and/or extent of encrustation and polymer contact angle. It is concluded that these IPNs may be of clinical benefit in patients providing stent resistance to extrinsic compression of the ureter in comparison with native PU. The comparable resistance to encrustation between the IPNs and PU indicates that the use of IPNs should not be restricted in this regard. PMID- 15348825 TI - Effect of dilution on the kinetics of cross-linking thermal polymerization of dental composite matrix resins. AB - The effect of dilution of 2,2-bis-(4-(2-hydroxy-3-methacryloxyprop-1oxy) phenyl) propane (bis-GMA) with triethylene glycol demethacrylate (TEGDMA) on the extent of polymerization, Ep, has been studied using differential scanning calorimetry. The isothermal and dynamic measurements indicate that the Ep is affected dramatically by dilution. The residual polymerization was also found to be strongly influenced by dilution. However, the apparent activation energy, Eapp, was found to be independent of dilution. PMID- 15348826 TI - Phenotypic characterization of keratinocytes migrated from polymer support - in vitro study. AB - The keratinocytes are able to migrate from the poly (2- hydroxyethylmethacrylate) disc if it is transferred to the new Petri-dish colonized with irradiated 3T3 mouse fibroblasts, and form a ring-shaped colony around the disc. The phenotypic characterization of human keratinocytes migrated from these discs was studied using a group of monoclonal antibodies. The keratinocytes in the external periphery of the colony of cells which migrated from the disc express the proliferating cell nuclear antigen (PCNA), alpha2, alpha3 chains and alpha5beta1 integrin receptor. A protein of the desmosome complex, desmoplakin-1, was also expressed. Involucrin and cytokeratin-10 were expressed after prolonged cultivation. These results suggest that the migrated keratinocytes are able to proliferate, recognize extracellular matrix molecules important in the process of the re-epitelization of the wound, and terminally differentiate in vitro. They are encouraging for further experiments with respect to the development of a support for keratinocyte cultivation and for grafting in clinical practice. PMID- 15348827 TI - Strengths of modified amalgam-to-dentin interphases. AB - Amalgam remains the primary dental restorative for nearly 200,000,000 Indonesians. Ground occlusal dentin surfaces of 140 formaldehyde-treated human molars were used to study shear strengths of different adhesive bonds (40: Syntac/Variolink, 40: Amalgambond Plus (AP), 40: AP+microfibre, and 20: Amalcoden) to a spherical high-copper amalgam (Valiant, Ivoclar NA). After 24 h at 37 degrees C/100 RH, restorations were stored for 7 d under one of four different conditions (all 37 degrees C) : A, 100% RH; B, deionized water; C, 0.9% NaCl solution; D, saliva electrolyte solution. Amalcoden-treated samples were exposed to conditions A and B only. ANOVA and Tukey post hoc analysis (P<0.05) were applied. For any single storage condition, AP+microfibre bond strengths were significantly greater than those for other agents tested. Storage condition was a significant variable only for the AP+microfibre; weaker bond strengths were observed for saline-stored samples. Improvement of restorative-to-dentin interfaces, eliminating pulp sensitivity, can result from aldehyde pre-treatment of teeth, as well as from the use of microfibre-filled Amalgambond Plus. These results illustrate the importance of "stabilization" of the entire interphase. PMID- 15348828 TI - Immobilization of calcium phosphate nano-clusters into alkoxy-derived porous TiO2 coatings. AB - Alkoxy-derived porous coatings of titanium oxide were fabricated on commercially pure titanium substrates by an electrochemical method in methanolic electrolytes. Nano-clusters of brushite (CaHPO4. 2H2O) were immobilized into the pores of the oxide network by reacting these coatings in acidic calcium phosphate solutions at 50 degrees C. The acid-base reaction between calcium phosphate solutions and the hydroxyl groups of the oxide network resulted in the formation of nano-clusters of brushite crystals immobilized inside the oxide pores. This treatment resulted in the conversion of the porous oxide network into a coherent mass with improved physical integrity. Nano-clusters of brushite crystals immobilized in the oxide matrix were converted into amorphous calcium phosphate (ACP) and poorly crystallized hydroxyapatite (HA) by further treatment of the oxide in alkaline solutions. The porous oxide coating also reacted strongly with concentrated phosphoric acid. The phosphate-modified oxide resulting from this reaction was further treated in calcium hydroxide solution to form nano-clusters of poorly crystallized HA within the oxide network. PMID- 15348829 TI - The influence of stress conditions on the wear of UHMWPE for total joint replacements. AB - In vitro studies of the effect of contact stress on the wear of ultra high molecular weight polyethylene (UHMWPE) in orthopaedic applications have produced contradictory results which predict both increased and decreased wear with increasing contact stress. In vivo studies of functioning hip prostheses have reported that 22 mm femoral heads generate lower linear and volumetric wear rates than 32 mm femoral heads. The effect of decreasing the head size will increase the contact stress but decrease the sliding distance per motion cycle. The present study consists of wear experiments under a range of contact stress magnitudes and application conditions in order to simulate the wear processes occurring in vivo. The results from these tests indicated that the wear factor actually decreases with increasing contact stress if the stress was not varied with time. If a time dependent or spatially varying stress was applied, the wear factor can increase greatly when compared to similar magnitude constant contact stress. This effect may be due to the complex relationship between the rate of wear particle generation and the rate at which the particles are released from the interface. The results of these wear experiments are discussed in terms of the influence of the stress conditions upon potential wear processes in total hip and knee prostheses. PMID- 15348830 TI - Study of interface phenomena between bone and titanium and alumina surfaces in the case of monolithic and composite dental implants. AB - The interface between mandibular bone and dental implants was examined with the in vivo dog model. Implant/bone interfaces were investigated for three types of materials: Ti-30 wt% Ta/Al2O3, titanium and Al2O3 using microscopy techniques covering a large magnification range: scanning electron microscopy, transmission electron microscopy, energy dispersive X-ray analysis and Auger spectroscopy. During the interaction of the Al2O3 ceramic with bone, an interfacial layer about 15 microm thick is formed. The same phenomenon was observed at the titanium bone interface, where the thickness of the layer was about 10 microm. In all cases, interface layers were sharp with well-defined borders between bone tissue and implant materials. No calcification took place inside the interface layer. A chemical analysis performed on this layer shows the presence of titanium, calcium and phosphorus in the case of titanium implants, and aluminium, calcium and phosphorus in the case of alumina implants. A rapid decrease in metal composition with increasing distances from the implant surface is correlated to a slow increase in calcium and phosphorus in the direction of the bone. Direct contact between implant and bone was observed. No biocorrosive effects were detected at the Ti-30 wt% Ta/Al2O3 metal-ceramic interface. PMID- 15348831 TI - Fourier-transform infrared spectroscopy study of an organic-mineral composite for bone and dental substitute materials. AB - A new injectable biomaterial for bone and dental surgery is a composite consisting of a polymer as a matrix and bioactive calcium phosphate (CaP) ceramics as fillers. The stability of the polymer is essential in the production of a ready-to-use injectable sterilized biomaterial. The purpose of this study was to detect possible polymer degradation which may have been caused by the interaction with the fillers using Fourier transform infrared spectroscopy. Composites containing CaP fillers (biphasic calcium phosphate, hydroxyapatite and peroxidized hydroxyapatite) and polymer (hydroxypropyl methyl cellulose) were prepared. To investigate the properties of the polymer, the inorganic and organic phases of the composite were separated using several extraction methods. The difficulty in separating the organic (polymer) from the mineral (CaP fillers) phases in the composite investigated in this study suggested the presence of strong interactions between the two phases. Spectra of extracted polymers showed new absorption bands of low intensities and indications that some chemical modifications of the original polymers have occurred. Results also indicated that the filler composition has an effect on the integrity of the polymer. PMID- 15348832 TI - Conditioning fluid influences on the surface properties of silicone and polyurethane peritoneal catheters: implications for infection. AB - Catheter-related infection remains a considerable problem in continuous ambulatory peritoneal dialysis (CAPD). This study examined the adherence of clinical isolates of Staphylococcus epidermidis to commercially available polyurethane and silicone peritoneal catheters in the presence and absence of a proteinaceous conditioning film. In addition, the effects of the conditioning film on the surface properties (advancing and receding contact angles, and surface rugosity) of these biomaterials were investigated. Bacterial adherence to polyurethane and silicone catheters, pre-treated with phosphate-buffered saline (PBS) or artificial spent peritoneal dialysate (ASD) for 1 h at 37 degrees C, was examined using a radiometric (2-3H-adenine) adherence assay. The advancing and receding contact angles and the surface rugosity of ASD- and PBS-treated biomaterials were examined using a dynamic contact angle analyser and an atomic force microscope, respectively. The bacterial isolates were selected to represent high and low cell surface hydrophobicity. The hydrophobic isolate exhibited both a significantly greater rate and a significantly greater extent of adherence than the hydrophilic isolate to both catheter materials, independent of pre-treatment. In general, pre-treatment of the catheter materials with ASD significantly decreased the subsequent adherence of both isolates owing to the deposition of a conditioning film on the surface of the biomaterial. ASD treatment also decreased both the advancing and receding contact angles and the surface rugosity of both catheter materials. This study highlights the influence of both bacterial cell surface hydrophobicity and biomaterial surface conditioning films on bacterial adherence to CAPD catheters. In addition, it is recommended that the effects of proteinaceous conditioning films on biomaterial surface properties should be considered when assessing materials for medical devices and products. PMID- 15348833 TI - Aluminablasting of human tooth enamel. AB - As part of a programme aimed at enhancing the degree of bonding that can be developed between dental polymer and human tooth enamel, the enamel has been subjected to two types of pretreatment, namely, aluminablasting (a mechanical etching) and aluminablasting followed by surface etching with orthophosphoric acid. Analysis of the surfaces so prepared by scanning electron microscopy has shown that enamel patterns with different degrees of retention ability have been obtained, and that the surfaces should be capable of bonding with dental polymer. PMID- 15348834 TI - Characterization of hydroxyapatite and carbonated apatite by photo acoustic FTIR spectroscopy. AB - An understanding of the interfacial relationship between a bioceramic implant and the adjacent bone tissue is facilitated by precise characterization of the associated structures. The structure of different commercial synthetic hydroxyapatite powders and a novel carbonated apatite have been studied with photo-acoustic (PAS) Fourier transform infrared (FTIR) spectroscopy. The PAS technique is an ideal method for analysing biomaterials, as materials can be analysed without the need to reduce the particle size or to dilute with KBr. Spectra from carbonated apatite appear to be different from those of commercial hydroxyapatite powders, with the main difference lying in the carbonate and phosphate ratio. Commercial hydroxyapatite powders from different sources have also been analysed and compared. PMID- 15348835 TI - In vitro biocompatibility of EPM and EPDM rubbers. AB - The in vitro toxicity of two EPDM rubbers (K 778 and K 4802) and one EPM rubber (K 740) was tested using human fibroblasts. The modulus of elasticity of each rubber was varied by exposure to different amounts of electron-beam radiation (0, 5 and 10 Mrad). The short-term in vitro toxicity was tested by culturing cells on polymer films. The long-term effect of ageing was simulated by growing fibroblasts in nutrient media prepared from extracts of heat-exposed materials. Cell cultures were studied both quantitatively and (ultra) structurally. Growth curves obtained in the toxicity test did not differ significantly from control values at any day of observation, and also showed that electron-beam radiation did not alter the biocompatibility. The same results were found for all but one material in the artificial ageing test. The number of cells in the K4802/10 Mrad extraction medium was decreased. Ultrastructurally no gross deviations from normal morphology were observed, either in the direct contact test or in the artificial ageing test. The most characteristic feature was a somewhat dilated endoplasmic reticulum. In summary, the in vitro biocompatibility of EPDM-rubbers as observed in this study is satisfactory and motivates further investigation of their biocompatibility in animal experiments. PMID- 15348836 TI - The optimum dose of gamma radiation-heavy doses to low wear polyethylene in total hip prostheses. AB - Wear volume, surface area and coefficient of friction of UHMWPE cup crosslinked with gamma radiation of 0, 50, 75, 100, 125, 150 and 200 Mrad sliding against an alumina ball were measured using a sphere-on-flat reciprocating type tribology testing machine. The effects of gamma radiation were scarcely observed in coefficient of friction. The coefficient of friction under lubricated (distilled water) and non-lubricated testing conditions was 0.08 to 0.12 and 0.20 to 0.25, respectively. The wear volume of UHMWPE with radiation of 50 Mrad, 75 to 150 Mrad, and 200 Mrad was 70 to 80%, 18 to 25%, and 12 to 15%, respectively, in comparison to non-irradiated specimens. Elongation and tensile strength of UHMWPE with radiation of 100 Mrad decreased to 6%, and 50% of that without radiation, respectively. The hardness increased with increase of the radiation dose. From several kinds of tribological findings, mechanical strength tests, and studies of long-term clinical findings, it is concluded that approximately 200 Mrad is the optimum dose of gamma radiation for clinical use in total hip prostheses. PMID- 15348837 TI - Comparative study of human monocyte and platelet adhesion to hydrogels in vitro - effect of polymer structure. AB - Blood platelet and monocyte adhesion was studied in vitro with respect to the influence of hydrophilic polymer chemical functional groups and their charge. The results showed that the strongest adhesion of human monocytes was to coverslips covered with cationic polymer. Platelet adhesion to all tested polymers proved to be negligible; no differences related to the charge of the polymers used were observed. These results show the obvious difference between the biocompatibility and haemocompatibility in vitro which must be taken into consideration during polymer biological properties testing before clinical trials. PMID- 15348838 TI - Preparation and characterization of thermally stable nanohydroxyapatite. AB - Rod-like hydroxyapatite was synthesized by precipitating calcium nitrate tetrahydrate and ammonium dibase phosphate in the presence of polyacrylic acid followed by hydrothermal treatment. Both polyacrylic acid and the hydrothermal treatment stabilised the hydroxyapatite. PMID- 15348839 TI - Sintering and characterization of HA and TCP bioceramics with control of their strength and phase purity. AB - HA and beta-TCP-based ceramics were prepared using commercial powders. Powder characteristics were defined and the processing parameters studied, aimed at the production of samples with improved microstructural and mechanical properties. The behaviour of HA powder subjected to various thermal treatments was investigated in order to control the formation of secondary phases (alpha- and beta-TCP) during sintering. The optimal thermal treatment required to prepare pure beta-TCP powder from the precursors (HA and DCP) was determined and the sintering method required to prepare fully dense beta-TCP completely free from alpha-form, was identified. Translucent hot-pressed beta-TCP ceramics with potential applications in aesthetic restorative prostheses were prepared and characterized. The interval of existence of alpha-TCP and alpha-TCP as secondary products was also defined. Crystallographic analysis was carried out on the imperfectly known low-temperature alpha-TCP phase, and a proper monoclinic unit cell determined. PMID- 15348840 TI - Synthesis of BISGMA derivatives, properties of their polymers and composites. AB - 2,2-Bis[4-(2 hydroxy-3-methacryloyloxy propoxy) phenyl] propane (BisGMA) is commonly the main component of the organic matrix of dental filling materials. Derivatives of BisGMA were synthesized from the diglycidyl ether of Bisphenol A (DGEBA) by the parallel reaction with methacrylic acid (MAA) and isophthalic acid as well as mixtures of methacrylic anhydride with palmitic acid and acetic anhydride, respectively, whereby MAA was partially substituted by the latter components. By this technique the structure of BisGMA monomer could be varied with regard to weight content of C=C double bonds, the hydrophilicity of the molecule as well as its flexibility or stiffness. Free-radical initiated homopolymerization of the monomers was carried out at 80 degrees C. Composites, prepared from mixtures of monomers with triethylene glycol dimethacrylate (TEGDMA) filled with 76% silica were room temperature polymerized using both redox and photoinitiated techniques. The polymerization shrinkage, diffusion coefficients of water in the crosslinked polymer, and some thermal properties of the homopolymers were determined. Mechanical properties of the resulting polymers and composites are compared to those of BisGMA itself. PMID- 15348841 TI - A study of cell behaviour on the surfaces of multifilament materials. AB - Since many fibres are very strong, they are considered to have useful potential for fibre reinforcement of orthopaedic and dental implant materials. Fibres exposed on the surface of composites may significantly influence the cellular response not only due to the chemistry but also due to the fibre size and shape. This study has concentrated on investigation of cellular responses to fibre specific aspects of fibre-reinforced composites. Four multifilament materials with diameter less than 20 microm were used: Kevlar 29(K), silicon carbide(SiC), nylon 66(N), and polyethylene terephthalate(PET). Established cell line L929 fibroblasts were used as the cell model. Cell behaviour on the surfaces of fibres was examined using direct cell counting (after 3, 5, 8 h and 1, 2, 3 days), scanning electron microscopy (SEM) (after 2 h and 2 days), and fluorescent staining of F-actin, which was analysed by confocal laser scanning microscopy (CLSM) (after 2 h and 2 days). The results showed that fibroblasts adhered and grew very well on all fibre surfaces, although less cells were observed on PET from direct cell counting. Significant orientational behaviour of cells was found on all fibre surfaces from the SEM and CLSM analysis, independent of the bulk chemistry of the fibres. PMID- 15348842 TI - Experimental analysis of tensile properties of some suturing materials. AB - The aim of the investigation is to quantitatively evaluate the failure load of several suture materials currently used in dentistry surgery. No chromic catgut, silk, Prolene, Ethilon, Ethibond, Vicryl and Vicryl Rapid, obtained from Ethicon s.p.a., in the sizes 3-0, 4-0, and 5-0 have been tested. The analysis has been carried out measuring the diameter of each suture with an optical microscope to determine the accuracy of manufacturers' data. Tensile testing has been performed to evaluate the failure load of each material. Finally, sutures of the same effective diameter class have been compared relative to failure load. Results show that monofilament sutures present a failure load remarkably superior to that of multifilament sutures. Using SEM analysis monofilament sutures present less surface irregularities than multifilament sutures. Only monofilaments Ethilon 4-0 and 5-0, Prolene 4-0, and multifilament no chromic catgut 4-0 and 5-0 meet the requirements of the Italian Pharmacopeia. In contrast, Prolene 5-0 and the other multifilaments, silk, Ethibond, Vicryl and Vicryl rapid, have a larger diameter than that declared on the label by the producer. PMID- 15348843 TI - Dental application of polyfunctional urethane comonomers to composite resin veneering materials. AB - Urethane monomer/diluent monomer mixtures were used in dental composite resin veneering materials filled with various ratios of powder (filler)/liquid (comonomer), P/L. Hardness values of unfilled resins containing benzoyl peroxide only (BP0; 0.5 wt%), and filled resins (included trimethylol propane trimethacrylate (TMPT) composite filler) were tested. Significant increases in hardness were obtained with the use of TMPT composite filler in the resins. Also, their modulus values measured by bend test showed an increasing trend, compared to a commercial composite resin veneering material (a control sample; CONT) with a lower filler content (50 wt%). The DME-DPMDC/HPDM comonomer (dimethacryloxyethyl diphenylmethane-4,4'-dicarbamate/hydroxypropyl dimethacrylate), which showed a smaller fraction of surface porosity, gave greater mechanical strength values at P/L ratios of 0.55 (17.8 wt% filler content) to 1.20 (27.3 wt%) than a CONT resin. The coefficient of thermal expansion was smaller in urethane-based filling materials than a CONT resin. Also, greater activation energy of thermal decomposition was observed in the resin samples with P/L ratio 0.75 to 1.20 than in a CONT resin. Thermally-induced decomposition occurred with smaller weight loss in the experimental filled resins than in a CONT resin. PMID- 15348844 TI - Bioactivity modulation of bioactive materials in view of their application in osteoporotic patients. AB - The application of bioactive ceramic coatings to prostheses confers strength to a material (ceramic or biological glass) that exerts beneficial effects on bone tissue growth but that itself lacks the toughness and stability required of an implant device. The rate of bioactivity is related to the chemical reactivity of the material and causes interface dissolution, precipitation and ion-exchange reactions. Ceramics may differ in sintering temperature and thus exhibit differences in their in vitro dissolution features and in vivo performance. To test these effects, in vitro and in vivo studies were carried out on two biocompatible biological glasses and a ceramic of proven bioactivity in view of their potential utilization as covering materials. In addition, a modified chitosan was adsorbed on the surface of a series of hydroxyapatite (HA) samples. Human fibroblasts and/or osteoblasts were used for the in vitro tests, and normal (INT) and osteoporotic (OVX) rats, normal rabbits and sheep for the in vivo studies. Similar chemical changes were observed in both glasses, suggesting that these materials underwent modifications directly dependent on their biological environment. The in vivo tests point to the possibility of improving the bioactivity of ceramic substrates with chitosan. However, the different behaviour of the materials in vitro and in vivo suggests that these tests should be conducted in parallel. PMID- 15348845 TI - Chemical characterization of in vivo aged zinc polycarboxylate dental cements. AB - The chemical composition of zinc polycarboxylate dental cements aged in vivo was studied. Thirty samples aged from one to 17 years were investigated using X-ray diffraction, infrared spectroscopy, thermogravimetric analysis and differential scanning calorimetry. Evidence for the presence of zinc oxide, amorphous zinc polycarboxylate and water of hydration was found. No correlation with age concerning either the chemical structure of the components or their relative amounts was found. Zinc polycarboxylate dental cements show very good chemical stability on long-term use. PMID- 15348846 TI - Predictive modelling of the mechanical properties and failure processes in hydroxyapatite- polyethylene (Hapex) composite. AB - The development of a wide range of hydroxyapatite polyethylene composites for medical applications is increasing the need for accurate predictive modelling. The objective of this work was to elucidate the observed mechanical processes and failure processes in this material using the finite element analysis method. The need for full three-dimensional modelling of this material has been shown. The results from this predictive model lead to accurate predictions of measured mechanical properties, and allow deduction of possible routes to improved ductility at high volume fractions. PMID- 15348847 TI - Interaction between self-assembled protein vesicles and microporous apatite surface. AB - Self-assembled structures such as vesicles have generated immense interest in recent decades due to their potential in mimicking biological membranes and in acting as drug-delivery systems. Despite the importance of the interaction between these organized assemblies and the surface of biomaterials, little is known about the mechanism involved. In this study, the interaction between giant proline-rich mussel adhesive protein (MAP) vesicles and the microporous apatite surface was investigated by scanning electron microscopy (SEM). We have found that MAP vesicles incubated on the apatite surface similar to osteoclasts, induce site-specific resorption of the apatite surface. However, in contrast to the osteoclastic resorption, the vesicle-induced resorption process appears to be accompanied by an organic matrix-mediated remineralization process. This results in the formation of a variety of complex three-dimensional site-specific "remodelled zones" on the apatite surface of micrometre scale. The mechanism of the formation of "remodelled zones" is discussed in terms of surface phenomena, such as adsorption and deformation of vesicles, site-specific release of resorptive agents, organic matrix-directed remineralization, and Ca-induced fusion, collapse and reshaping of the vesicles on the apatite surface. PMID- 15348848 TI - Bone remodelling in the pores and around load bearing transchondral isoelastic porous-coated glassy carbon implants: experimental study in rabbits. AB - Cylinders of porous-coated glassy carbon were implanted into drill holes made through the articular surface of the medial condyle of both tibiae of ten rabbits for six and 12 weeks. Bone ingrowth and remodelling was examined by radiographic, histologic, oxytetracycline-fluorescence and microradiographic methods. Bone ingrowth into pores and load bearing implants was seen by all examination methods. Bone ingrowth occurred earlier when the pores were facing cancellous bone than cortical bone. Appositional bone formation occurred on the trabeculae a few millimetres from the interface during the early phase of remodelling at six weeks. At 12 weeks resorptive remodelling had occurred both in the surroundings and in those pores that face cancellous bone, whereas the amount of bone still increased in the pores facing cortical bone. In its porous-coated form glassy carbon functions well as a frame for ingrowing bone and it shows good osteoconductivity. Its mechanical properties are suitable for functioning as a structural bone substitute in places where the loads are mainly compressive. The difference between findings at six and 12 weeks indicated physiologic stress distribution and the adverse effects of stiff materials on bone remodelling were avoided by using this isoelastic material. PMID- 15348849 TI - Comparison of the wet and dry fatigue properties of all ceramic crowns. AB - The purpose of this paper was to investigate the influence of fatigue on the fracture strength of In ceram, optimal pressable ceramic (OPCTM) and IPS Empress in both wet and dry environments. Twenty-six crown shapes 8 mm in diameter and 8.5 mm in height were fabricated for each ceramic system. For each ceramic system, ten specimens were tested for fracture strength without fatiguing. The second group was submitted to a fatigue and fracture test in a dry (eight specimens) and a third group in a wet (eight specimens) environment using an Instron testing machine. The results were statistically analysed using a Mann Whitney test. The results indicated that: (i) the fracture strength for In ceram was significantly stronger than OPCTM and IPS Empress (p<0.05) - no difference was found between OPCTM and IPS Empress; (ii) fatiguing and fracture testing showed a significant decrease in the fracture strength for In ceram and IPS Empress in the wet environment and no difference was found in the dry environment - no difference was found for OPCTM; and (iii) when fatigued in a dry environment, In ceram crown shapes were significantly stronger than OPCTM and IPS Empress (p<0.05) - the same statistical differences were found when fatigued in a wet environment. PMID- 15348850 TI - Evaluation of the tissue reaction to a percutaneous access device using titanium fibre mesh anchorage in goats. AB - The tissue reaction to a percutaneous access device, applicable as a carrier for an implantable glucose sensor, was evaluated in goats. Titanium fibre mesh structures were used for anchorage of the device in superficial as well as deeper soft-tissue locations. The percutaneous part was subcutaneously anchored with a fibre mesh sheet. The distal part was placed intraperitoneally and anchored in deeper soft-tissue layers using a fibre mesh cuff. All implants showed good healing with the surrounding tissue. Histological evaluation showed that the subcutaneous fibre mesh sheets and peritoneal fibre mesh cuffs were filled with immature connective tissue, generally free of inflammation. Problems concerning disconnection of the silicone catheter from the titanium holding element and filling of part of the peritoneal fibre mesh cuff with silicone glue have to be overcome by more appropriate preclinical testing and improved implant design. Our results demonstrate that titanium fibre mesh structures can be used effectively for soft-tissue anchorage of percutaneous access devices. A sufficient ingrowth of connective tissue was obtained in superficial as well as in deeper soft-tissue layers. The access device could have application as a carrier for an implantable glucose sensor for glucose monitoring in different tissue compartments. PMID- 15348851 TI - Investigation of glass-ionomer cements using differential scanning calorimetry. AB - Six commercial glass-ionomer cements commonly used for various dental applications have been investigated using differential scanning calorimetry (DSC). The heat-flow behaviour and heat capacity of the cements were measured during isothermal (at 37 degrees C) setting reactions. The DSC results show that all materials undergo an exothermic setting process, but with different enthalpies of reactions and different heat capacities; there are no remaining endo- or exothermic reactions after the setting of the cement. All materials examined were found to be effective thermal insulators. PMID- 15348852 TI - Studies of the healing of bone grafts, and the incorporation of titanium implants in grafted bone: an experimental animal model. AB - An insufficient quality and amount of bone often necessitate the clinical use of implants together with bone transplants. The present study describes an experimental animal model for the study of implants in bone grafts. Circular defects were made bilaterally in the tibia of 36 rabbits. The defects received either autologous cortical bone (control), demineralized bone matrix (DBM), plasma-augmented DBM or were left empty (without bone graft). In all defects a titanium implant was centrally placed and anchored in the opposite cortex. Evaluation with light microscopic morphometry showed that the insertion of a threaded titanium implant centrally in a cortical defect was followed by a spontaneous healing of the defect after 6 mon. After 6 wk, all implants in cortical grafts were well integrated with a significantly higher bone-to-implant contact than in the DBM and plasma-augmented groups. After 6 mon, all experimental groups had a mean bone area within the threads ranging between 69% and 80% and a mean bone-to-implant contact between 31% and 42%. The results from the present study indicate that the model allows comparative studies on the early formation, resorption and remodelling of bone around implants after modification of implant, graft and host properties. PMID- 15348853 TI - In vitro testing of surface-modified biomaterials. AB - The influence of surface modification treatments such as ion implantation and sputter coating on an in vitro rat bone-marrow cell culture was studied by scanning electron microscopy and X-ray microanalysis. 316 L stainless steel, Ti 6Al-4V and Ti-5Al-2.5Fe were nitrogen ion-implanted with three fluences: 10(15), 10(16) and 10(17) ion cm-2 with an energy beam of 40 keV. Both nitrogen and carbon sputter-coated 316 L stainless steel samples were also studied. Polished 316 L stainless steel, Ti-6Al-4V, Ti-5Al-2.5Fe and Thermanox were also studied, in order to give comparative information. The materials were inoculated with a droplet of cell suspension and were maintained for 3 wk. A mineralized extracellular matrix was formed on all materials except on nitrogen sputter coated 316 L stainless steel. The morphology of the cell cultures obtained on nitrogen-ion implanted materials was similar to those obtained on the untreated materials and Thermanox. The observation of the interface between the cell layer and the substrata showed the presence of calcium- and phosphorus-rich globular deposits associated with collagen fibres. A higher density of these globular deposits was observed on the ion-implanted materials. PMID- 15348854 TI - Some factors controlling the injectability of calcium phosphate bone cements. AB - The injectability of four calcium phosphate bone cements (CPBCs) was measured using a commercial disposable syringe. It varied considerably with the cement powder composition, with the liquid/powder ratio, with the time after starting the mixing of liquid and powder, with the accelerator concentration (% Na2HPO4), and with the ageing time of the cement powder which was prepared by milling. The injectability test could be used to determine accurately the dough time of CPBCs. Relations between the setting time and the cohesion time are discussed. PMID- 15348855 TI - A comparison of the corrosion behaviour and surface characteristics of vacuum brazed and heat-treated Ti6Al4V alloy. AB - The corrosion characteristics of the brazed Ti6Al4V specimens were analysed and compared with respect to the conventionally heat-treated specimens by an electrochemical corrosion test. The object of this research was to explore the potentiality of the brazed titanium for biomaterials. The characteristics of the 1300 degrees C heat-treated and the 970 degrees C brazed specimens, with passivation and sterilization treatment, were evaluated by measurement of corrosion potential, Ecorr, corrosion current densities, Icorr, polarization resistance of the reacted surface films, Rp, in a potentiodynamic test. The experimental results show that the corrosion rates of the heat-treated and the brazed samples are similar at Ecorr, and the value of Ecorr for the brazed sample is noble to the heat-treated samples. The passive current density of the brazed specimen is either lower or higher than the heat-treated specimen, depending on the polarization potential. By Auger electron spectroscopic and high-resolution X ray photoelectron spectroscopic analysis on specimens from the potentiostatic test, the elements of copper and nickel in the brazing filler were not detected while less alumina was found in the reacted film of the brazed specimens when compared with the heat-treated specimens. The implication of the results is discussed. PMID- 15348856 TI - Surface characteristics of Ti6Al4V alloy: effect of materials, passivation and autoclaving. AB - The properties of passivated films for Ti6Al4V alloy prepared by various methods (as-polished, brazed at 970 degrees C for 2 h and brazed at 970 degrees C for 8 h) were investigated. Four passivations (non-treated, nitric acid passivation, 400 degrees C-treated in air, and aged in boiling water), with or without autoclaving treatments, were adopted for evaluating the changes of surface properties, including chemical composition, chemical structure, and oxide thickness. From X-ray photoelectron spectroscopic (XPS) analyses, surface elements of copper and nickel in brazed samples were undetected for non-treated, acid-passivated and boiling water-aged specimens, while they were found in the 400 degrees C-treated specimen. The relative contents of Ti2++Ti3+ to Ti4+ were determined by passivation treatments, but were not related to the experimental materials and autoclaving treatment. Passivation and autoclaving decreased the Ti to Ti4+ ratio by virtue of an increase in oxide thickness. Of the four types of passivation treatment, the 400 degrees C thermal treatment exhibited the lowest content of suboxides and metallic elements and the thickest oxide by XPS analysis; however, this treatment may cause a desorption of the basic OH group in the hydration layer on the surface of titanium alloy. PMID- 15348857 TI - The polymer Polyactive as a bone-filling substance: an experimental study in rabbits. AB - The biocompatible, osteoconductive and resorbable polymer Polyactive (PA) was investigated for its performance as a bone-graft substitute. The model consisted of a 4 mm borehole, 1.5 cm distal of the major trochanter in both femurs of a rabbit, of which one was filled with a cylinder of porous PA. The other was left untreated. PA70/30 and PA60/40 were investigated, both before and after being incubated with allogenic bone marrow. Analyses were performed after 4, 8, 26 and 52 weeks and comprised dual energy X-ray absorptiometry (DXA) and image analysis of histological sections. DXA revealed an increased bone mineral density in the filled defects compared to the controls, both at the defect and immediately proximal and distal of the defect. Histology showed that gap-bridging had occurred within 8 weeks, with 80%-90% of the pores of PA70/30 and PA60/40 occupied by new bone, and an intimate bone-PA contact. PA70/30 seemed to be more suitable compared to PA60/40, in that the highest amount of bone was formed within the shortest period of time. Incubation of PA with allogenic bone marrow resulted in inflammatory reactions at the sites of implantation, which delayed bone growth, but did not prevent it. It was concluded that PA70/30 and PA60/40 are suitable bone-graft substitutes. PMID- 15348858 TI - Application of laminates to mouthguards: finite element analysis. AB - A finite element model comprising a flat-ended indentor and a disc representing a colliding object and mouthguard materials, respectively, has been developed to study stress distribution and impact force in laminates. The disc consists of two layers and its top layer is in contact with the indentor. Two different combinations of layers were employed for the simulation. One had a soft layer placed on top of the rigid layer and the other was vice versa. It was found that the former had no significant difference from a monolayer in stress distribution and impact force. However, the latter was found to have a significant effect on stress distribution, and this effect could be increased by controlling ratios of modulus and volume fractions of the top and bottom layers. It was also found that the magnitude of the impact force increases with increasing effect of stress distribution, but this competition can be reduced to some degree by decreasing the volume fraction ratio of top to bottom layers. PMID- 15348859 TI - Morphological and mechanical study on the effects of experimentally induced inflammatory knee arthritis in rabbit long bones. AB - Inflammatory knee arthritis was induced by intraarticular injection of carrageenan twice a week for a total of 6 weeks in New Zealand White rabbits and the effects of the arthritis on the morphological and mechanical properties of the adjacent femur and tibia were evaluated 8 weeks after the first injection. Carrageenan-induced knee arthritis resulted in severe osteopenic changes and a dramatic decrease in bone strength of the entire ipsilateral femur and tibia, including the femoral head and distal tibia, but not the contralateral femur and tibia and the remote humerus. The osteoporotic changes of the adjacent bones of the inflammatory arthritic knee are the basis for the reduced mechanical strength of these bones. These findings may have clinical significance with regard to the mechanisms and consequences of osteoporotic changes in patients with rheumatoid arthritis. PMID- 15348860 TI - The effects of polyethylene cup thickness on wear of total hip prostheses. AB - The wear rate of the ultrahigh molecular weight polyethylene cups in combination with a 28 mm alumina femoral head was measured on the radiographs of patients without any complications, and on cups retrieved due to slight loosening of the prosthesis or due to late infection between bone and components. The wear rate on the radiographs did not include the initial wear, but the wear rate on the retrieved cups did include the initial wear. The wear rate on the retrieved cups was higher, by 50%, than that measured on the radiographs. In both cases, the thicker the polyethylene cups, the lower was the wear rate measured. The average wear rate of the 7 and 8 mm thick cups was about twice that of cups 10 and 11 mm thick. From these results, we conclude that polyethylene cups more than 11 mm thick should be used. PMID- 15348861 TI - Ultrasonic implantation of calcium metasilicate glass particles into PMMA. AB - Polymer materials for clinical applications should be bioactive and have a bone bonding ability. In order to provide poly(methyl methacrylate) (PMMA) with bioactivity, granules (<45 microm) of a bioactive glass 50CaO.50SiO2 (mol %) were implanted into PMMA: they were suspended together with a piece of PMMA in a 40 tetrahydrofuran-60 ethanol (vol %) solution and ultrasonically agitated. The granules of <10 microm in size were impregnated at approximately 40-20 microm depth below the substrate surface. Two types were detected on the PMMA surface: (a) a glass-granule layer on PMMA, and (b) an inner granule layer, a PMMA layer, and an outer granule layer on the PMMA. The bioactivity of the implanted PMMA substrates was examined in vitro with a simulated body fluid (Kokubo solution). Apatite was precipitated on all glass granules and the whole substrate surfaces within 1 d. After 4 h soaking in the Kokubo solution, aggregates of apatite particles appeared on the substrate surface, independently of those on the glass granules, and they grew and proliferated on the whole subtrate surface in 7 d. Silica gel islands on PMMA due to the silicate anions from the glass were considered to induce nucleation of the apatite particles. PMID- 15348862 TI - Coating of CaTiO3 on titanium substrates by hydrothermal reactions using calcium ethylene diamine tetra acetic acid chelate. AB - Titanium plates were treated in [Ti(O2)EDTA]2- -Ca(EDTA)2- mixed solutions and/or Ca(EDTA)2- solutions (where EDTA is ethylene diamine tetra acetic acid) at pH 9 13 and 150-250 degrees C for 0.5-12 h. The film, about 50 microm thick, and consisting of mixtures of CaTiO3 and TiO2 was formed in 0.01 M [Ti(O2)EDTA]2- - 0.01 M Ca(EDTA)2- mixed solution at pH 13 and 250 degrees C for 6 h. The film consisted of large icosahedral and hexagonal particles, of about 10 microm diameter, and small aggregated particles, of about 1 microm diameter. On the other hand, the film, about 20 microm thick, consisted of hexagonal plate-like CaTiO3 particles, of about 1 microm diameter, was formed in 0.01 M Ca(EDTA)2- solution at pH 13 and 250 degrees C for 6 h. The thickness of both films increased with time, where the film formation rate in 0.01 M [Ti(O2)EDTA]2- - 0.01 M Ca(EDTA)2- mixed solution was much faster. The CaTiO3 film formed on the surface of titanium promoted the precipitation of hydroxyapatite on the substrate by the hydrothermal reactions in Ca(EDTA)2- -PO(4)3- mixed solutions. PMID- 15348863 TI - The effect of biological environment on the surface of titanium and plasma sprayed layer of hydroxylapatite. AB - The solubility of titanium samples with different surface coatings, i.e., hydroxylapatite (HA) powders, a two-layer coating of ZrO5+HA on a titanium substrate in solution and of tooth implants after long-term functioning in the human organism, was studied. A minimum difference in solubility of titanium samples with different surface finishes (polished or grit blasted) was established. For the HA powders and coatings, the lowest solubility was observed with a coarse-grained HA-B powder and a coating made of that powder. Clinical tests of tooth implants after long implantation times were performed. A titanium implant (implantation 12 y), a titanium implant with a two-layer coating of ZrO5+HA-A (implantation time 4 y) and a titanium implant with a two-layer coating of (Al5O3+3% TiO2)+HA-A (implantation time 6 y) were studied. The results show that the titanium surface and HA-A layers were dissolved. Nevertheless, after 6 y implantation, total removal of HA-A coating from that part of implant set into the bone, was not observed. PMID- 15348864 TI - Evaluation of copolymers of polyethylene oxide and polybutylene terephthalate (polyactive): mechanical behaviour. AB - Polyether-polyester segmented block copolymers (Polyactive) on the basis of polybutylene terephthalate (PBT) and polyethylene oxide (PEO) were mechanically tested. Tensile strength and modulus of elasticity in compressive and tensile deformation were recorded according to ASTM standards. These tests were done in vitro under dry and wet conditions, and after 3, 9 and 25 wk subcutaneous implantation of these materials in goats. Strength and modulus of elasticity were higher with increased contents of PBT in the copolymers. After water uptake, the polymer displayed a lower strength and stiffness. Disintegration of the materials with 70% PEO content and dumb-bell shape was noted at 3 wk. Disintegration of the cylinders of the same material was seen after 25 wk implantation. Of the materials with 60% PEO content, only four of the five dumb-bells had disintegrated after 25 wk implantation. The in vivo test results of all other implants did not show a clinically relevant decrease of strength and stiffness with time after implantation of the copolymers in the goats. Mechanical behavior of the various copolymers seemed mainly determined by the amount and integrity of the PBT phase. PMID- 15348865 TI - Influence of surface microstructure on the reaction of the active ceramics in vivo. AB - When porosity and macro-pore size differ in the same ceramic, the mode of bone regeneration and the degradation of ceramics in vivo is said to be different. However, the reactions in vivo of ceramics that have a different microstructure with the same porosity and the same macro-pore size, are not so far known. In this study, two kinds of beta-tricalcium phosphate (TCP) that had different microstructures but the same porosity and macro-pore size, were manufactured. These TCP were implanted in the distal femurs of 20 mature male rabbits, and their respective areas of ceramics and of regenerated bone were measured after 4, 12 and 24 wk. In both TCPs, the regenerated bone similarly decreased from 4-24 wk in a different way. The area of ceramics in one of these TCPs significantly decreased gradually throughout the observation period. On the other hand, the other TCP showed no marked decrease during the same period. This suggested a possibility that the difference in micro-structure has a large effect on the reaction of the ceramics in the bone. PMID- 15348866 TI - Formation of carbonate apatite on calcium phosphate coatings containing silver ions. AB - The prerequisite for bioactive materials to bond to living bone is the formation of biologically equivalent carbonate apatite on their surfaces in the body. Calcium phosphate ceramic surfaces can be transformed to a biological apatite through a series of surface reactions including dissolution-precipitation and ion exchange. In the present work, apatite coatings with different crystallinity, compositions and crystal sizes, including a well-crystallized hydroxyapatite coating, were synthesized electrochemically and doped with silver ions in silver nitrate solution at room temperature. The formation of a new carbonate apatite on the surface of these coatings was investigated in an acellular simulated body fluid with ion concentrations comparable with those of human blood plasma, using scanning electron microscopy and Fourier transform-infrared spectroscopy. The results show that small quantities of silver ions incorporated into apatite coatings may have a strong stimulatory effect on the formation of carbonate apatite without adversely affecting the chemical stability of these coatings. PMID- 15348867 TI - Retrieved total hip prostheses. Part I: the effects of cup thickness, head sizes and fusion defects on wear. AB - The effects of polyethylene cup thickness, femoral head sizes (22, 28 and 32 mm) and fusion defects on wear were investigated on retrieved total hip prostheses. When the cup thickness was less than 9 mm, the larger the femoral head, the higher the linear wear rate; however, when the cup thickness was more than 9 mm, the larger the femoral head, the linear wear rate was lower. When the cup thickness was less than 11 mm, the volumetric wear rate increased with increasing size of the femoral head, and when it was more than 11 mm, the volumetric wear rate of the three kinds of the prosthetic cups approached the same values. The wear rate of the cross-linked cups irradiated by 100 Mrad were very low, with no correlation to cup thickness. When the cup thickness was less than 9 mm, the volumetric wear rate tended to increase with increasing number of fusion defects. Large diameter fusion defects diminished the tensile strength. PMID- 15348868 TI - Bioactive glass granules and polytetrafluoroethylene membrane in the repair of bone defects adjacent to titanium and bioactive glass implants. AB - An experimental animal model was used to investigate the effect of bioactive glass (BG) granules and nonresorbable polytetrafluoroethylene (PTFE) membrane on the repair of cortical bone defects adjacent to titanium and BG implants. Thirty two Astra(R) (diameter 3.5 mm) dental implants were inserted bicortically and 42 conical BG implants (diameter 2.5-3.0 mm) monocortically, into fitted holes of rabbit tibia. Before implantation, a standardized bone defect was created by drilling an extra hole (diameter 3.0 mm) adjacent to each implant site. Twenty eight defects were filled with BG granules (diameter 630-800 microm) (BG group) and 28 defects were left empty but covered with PTFE membrane (PTFE group). No material was used in 18 control defects (control group). Morphometrical evaluation with a digital image analysis system was used to measure bone repair as percentages of the defect area on scanning electron microscopy (SEM) and light microscopy pictures. Bone-implant contact was measured as percentages of the thickness of the cortical bone. At 6 and 12 wk, bone repair in defects in connection with titanium implants was 23.2% and 36.6% in the BG group, 23.2% and 32.4% in the PTFE group, and 47.2% and 46.2% in control defects. Corresponding figures for BG implants were 33.2% and 40.1% in the BG group, 16.6% and 33.5% in the PTFE group, and 25.7% and 54.9% in control defects, BG granules and new bone together filled 82.7% and 68.5% of the defect area adjacent to titanium implants, and 75.9% and 74.4% of the defect adjacent to BG implants at 6 and 12 wk, respectively. Better bone-implant contact was achieved at the defect side with BG than titanium implants (77.0% versus 45.0% at 12 wk). The results indicate that BG granules are useful in treatment of bone defects adjacent to dental implants. BG coating of the implant seems to improve osseointegration in the defect area. PMID- 15348869 TI - Complement activation by cellulose: investigation of the effects of time, area, flow rate, shear rate and temperature on C3a generation in vitro, using a parallel plate flow cell. AB - The development and utilization of a parallel plate flow system to study the blood response to flat sheet biomaterials, is described. Unlike most other parallel plate flow systems, which have been used to study cellular interactions with biomaterials, the controlled flow test cell described below employs the test materials on both sides of the channel through which the blood flows. The flow cell is used to conduct an investigation into the in vitro generation of C3a by a regenerated cellulose membrane, Cuprophan. The effects of experimental variables such as temperature, blood flow rate, contact area and wall shear rate on C3a generation by Cuprophan were studied. The results show that C3a generation by Cuprophan is lower at 12 degrees C than at 22 degrees C, which is in turn lower than C3a generation at 37 degrees C. Furthermore, a decrease in contact area, and increase in wall shear rate and blood flow rate, can produce a decrease in C3a concentration. PMID- 15348870 TI - Analysis of the inflammatory response to titanium and PTFE implants in soft tissue by macrophage phenotype quantification. AB - Implants of commercially pure titanium and polytetrafluoroethylene (PTFE) were inserted in the rat abdominal wall for 1, 6 or 12 wk. The foreign body reaction was evaluated by immunohistochemical quantification of monocytes/macrophages and by the thickness of the foreign-body capsule. At all time intervals, the majority of interfacial cells were ED1-positive while ED2-positive cells were localized deeper in the tissue. Neither titanium nor PTFE displayed a significant change in capsule thickness over time. The total cell numbers decreased overtime for both types of material. At 12 wk the PTFE implants, compared to titanium, were surrounded by a significantly thicker reactive capsule with larger total cell numbers. No significant differences were seen in the macrophage subset response between the two types of implants. Thus, the present study showed differences between titanium and PTFE at 12 wk but not at earlier time points. PMID- 15348871 TI - Calcification of porcine and human cardiac valves: testing of various inhibitors for antimineralization. AB - Despite distinct advantages over mechanical cardiac valve prostheses, the use of bioprosthetic valves remains limited, due to poor long-term durability, primarily as a result of tissue calcification. A novel in vitro, anticalcification process based on treatment of porcine and human heart valves with metallocene dichlorides, as well as with amino acids (phenylalanine, aspartic acid, glutamic acid), has been developed. This anticalcification process reduced mineralization of porcine and human valves up to 32% and 28%, respectively. PMID- 15348872 TI - Attachment kinetics, proliferation rates and vinculin assembly of bovine osteoblasts cultured on different pre-coated artificial substrates. AB - Primary bovine osteoblasts were used to study in-vitro effects of attachment on vinculin assembly in cells cultured on various artificial substrates. Materials coated with fibronectin and bovine serum albumin (BSA) as well as untreated materials (tissue culture polystyrene and Aclar foils) were chosen to investigate substrate-dependent proliferation during the first 3 days of culture. Proliferation was highest on fibronectin-coated substrates, followed by BSA coated and untreated substrates. During the first 24 h of cultivation, cell attachment kinetics revealed no significant difference between the various substrates. After 24 h detachment rates obtained by calcium depletion with ethylenediaminetetraacetic acid were highest on uncoated materials, followed by BSA- and fibronectin-coated substrates. Phase contrast microscopy revealed typical osteoblast morphology after cell adhesion for 24 h. The dynamic attachment process was concomitant with the reassembly of vinculin into streak like focal contacts clustered on the ventral side of cells. The kinetics of vinculin reassembly were independent of the underlying coating. Thus, fibronectin coating of artificial substrates increased the attachment strength and proliferation rate of osteoblasts. While the reassembly of vinculin in focal contacts seems to be a prerequisite of osteoblast attachment in vitro, it does not seem to have profound effects on the subsequent cell behaviour on artificial substrates. PMID- 15348873 TI - Coating of ZrO2 supports with a biological glass. AB - The possibility of obtaining a good bioactive coating on biomedical devices made with zirconia ceramic was considered. Yttria partially stabilized zirconia was used to produce suitable substrates. The obtained adhesion was good (66+/-13 MPa) and encouraging for biomedical applications. Small microcrystals formed inside and on the surface of the glass layer. These microcrystals transformed the glass into a glass-ceramic. An accurate investigation of the nature of this microcrystal formation was carried out to verify its compatibility with the planned biomedical applications. PMID- 15348874 TI - Isothermal and non-isothermal polymerization of a new bone cement. AB - A new bone cement based on poly(ethylmethacrylate) (PEMA), hydroxyapatite powder (HA) and n-butylmethacrylate monomer (n-BMA) has been studied using isothermal and non-isothermal polymerization. Methacrylate monomers are highly reactive and release a considerable amount of heat during polymerization. A quantitative understanding of the methacrylate polymerization is necessary because the thermal history of the polymerization has considerable influence on the final properties of a bone cement. In the first part, polymerization kinetics are analysed by means of differential scanning calorimetry (DSC). DSC data are used to evaluate a phenomenological model describing the cure kinetics of this new bone cement. In the second part, a kinetic model coupled with the energy balance is used to obtain temperature and degree of conversion profiles in the bone-cement prosthesis system, under non-isothermal conditions, as function of initial temperature and thickness of the cement. Material properties, boundary and initial conditions and the kinetic behaviour are the input data for the numerically solved heat-transfer model. The temperature at the bone/cement interface, can be considered as a weak point, often responsible for total joint replacement failure. For this particular bone cement exhibiting a low exotherm and low glass transition temperature, the interfacial temperature is lower than the threshold level for thermal tissue damage (50 degrees C). The conversion occurs almost completely, avoiding problems with unreacted monomers that can be released by the cement, giving rise to tissue damage. PMID- 15348875 TI - Effect of fluoride on apatite formation from Ca4(PO4)2O in 0.1 mol L(-1) KH2PO4. AB - The effect of fluoride on the hydrolysis of tetracalcium phosphate (TTCP; Ca4(PO4)2O) was investigated in 0.1 mol l(-1)KH2PO4 containing 0-83 mol l(-1) KF. Characterization of the final apatite phase formed by the hydrolysis was made with X-ray diffraction and SEM. The initial pH was between 4.5 and 5.4, depending on the solutions, and the pH rapidly increased and was kept constant between 7.3 and 6.5. An increase in KF concentration tended to lower the pH in the final stage of hydrolysis. The calcium concentration was considerably lower than the phosphorus concentration throughout the reaction. The fluoride concentration decreased shortly after the start of hydrolysis. The hydrolysis of TTCP in 0.1 mol l(-1) KH2PO4 proceeded to form hydroxyapatite via DCPD when the KF concentration was low. The hydrolysis product was a calcium-deficient non stoichiometric hydroxyapatite with a Ca/P ratio of about 1.5. With an increase in the KF concentration in the 0.1 mol l(-1) KH2PO4 solution, TTCP directly transformed into hydroxyapatite containing F- ions or fluorapatite and with improved crystallinity. The addition of fluoride in the solution initially accelerated the formation of apatite. However, the layer of newly formed apatite adhering to the TTCP particles retarded TTCP dissolution; as a result, hydrolysis was delayed. IR analysis showed that the apatite phase contained HPO2-4 ions in the structure. The formula for the hydrolysis product of TTCP in the presence of fluoride can be expressed as follows: Ca10-x(HPO4)x(PO4)10-x(OH)2-x-yFy. PMID- 15348876 TI - Experimental evaluation of a percutaneous injectable biomaterial used in radio interventional bone-filling procedures. AB - Interventional radiology is beginning to be used in bone pathology. An injectable biomaterial Ethibloc*, composed of alcohol and a vegetal protein (zein), has been successfully used for percutaneous treatment of benign bone lesions. The reasons for this success remained unknown and needed to be understood. In this study, using a rabbit model, an evaluation was made of bone formation and tissue reaction during the first weeks after the injection of this biomaterial. Ethibloc* was injected percutaneously into bone defects in rabbit distal femurs. Three time intervals were studied: 2, 4 and 8 wks. The three control groups constituted unfilled, polymethylmethacrylate (PMMA), and alcohol. Undecalcified bone technique was used for a qualitative analysis and histomorphometric evaluation. A low bone formation was found which was less than in the control groups (PMMA and unfilled). The "Ethibloc group" was characterized by an early inflammatory reaction. Good clinical results obtained with Ethibloc* probably arose, after an initial vascular thrombosis, from a secondary bone reaction and spontaneous osteogenesis obtained after the disappearance of vascular or hyperpression reaction. PMID- 15348877 TI - In vitro investigation of novel calcium phosphates using osteogenic cultures. AB - A rat bone marrow stromal cell (RBM) culture was used to evaluate novel bioactive calcium phosphate ceramics. Three rapidly resorbable, glassy crystalline materials with the main crystalline phase Ca2KNa(PO4)2 were investigated (sample code GB 1a, GB 14, GB 9). These materials were designed to exhibit a higher degree of biodegradability than tricalcium phosphate. Additionally, a bioactive glass ceramic of low biodegradability was examined (sample code AP 40). RBM cells were cultured on the disc-shaped test substrata for 14 d. The culture medium was changed and calcium and phosphate concentrations of the medium were determined daily. Specimens were evaluated using light microscopy and morphometry of the cell-covered substrate surface, scanning electron microscopy and energy dispersive X-ray analysis. Except for GB 1a, the rat bone marrow cells attached and grew on all substrate surfaces. Of the different calcium phosphate ceramics tested, AP 40 facilitated osteoblast growth and the elaboration of the extracellular matrix to the highest degree followed by GB 9 and GB 14. The inhibition of cell growth encountered with GB 1a seemed to be related to its high phosphate ion release. PMID- 15348878 TI - Structure and strength of low-mercury dental amalgams prepared with liquid Hg 47.4% In alloy. AB - Two low-mercury amalgams: (1) low-copper lathe-cut and (2) high-copper (Tytin) were prepared by amalgamation with liquid Hg-47.4% In alloy. The strength structure relationship of these amalgams was investigated and compared with standard amalgams (i.e. amalgams prepared with the same powders and pure mercury). The matrix phase of the low-mercury amalgam was found to be depleted of mercury and may be thought of as In4Ag9 compound with some mercury dissolved, indicating that less mercury (compared with standard amalgam) combines with silver, thus producing a strong amalgam matrix. On the other hand, an increase was observed in the consumption of the initial gamma (Ag3Sn)-phase, leading to an increase of the tin released. As a result, the potential of [HgSn]-phase formation in low-mercury amalgams increases. The observed increase in the quantity of gamma2(Sn7-8Hg)-phase in low-copper amalgam, or its appearance in high-copper amalgam (where it is normally absent), contributes to a deterioration in the strength of the investigated amalgam. The conclusion drawn was that low mercury amalgam may be prepared with liquid Hg-47.4%In alloy but, in order to eliminate gamma2-phase formation, novel and possibly tin-free amalgamable alloys should be developed. PMID- 15348879 TI - Nanoporous alkoxy-derived titanium oxide coating: a reactive overlayer for functionalizing titanium surface. AB - Effective immobilization of bioactive substances such as adhesive proteins, synthetic peptides and growth factors on metallic substrates is required for a number of medical applications. In the present work, evidence is presented to show that an alkoxy-derived nano-porous titanium oxide coating, synthesized electrochemically on titanium in methanolic electrolytes, may act as an effective interface for functionalizing a titanium surface. It is demonstrated that nanoporous oxide coatings could facilitate fast diffusion of small organic molecules within the oxide network and form strong chemical bonds with the functional groups of these molecules at room temperature. Fourier transformed infrared spectroscopy was used to investigate the nature of the interfacial interactions between the oxide network and a range of molecules containing COOH, OH, NH2, C=O and phosphoric acid functional groups. The results indicate that the nanometre-sized oxide clusters within the coating may play an essential role in effective immobilization of organic molecules by providing numerous binding sites for chemisorption of these species. The surface-derivatized oxide coating may provide a solid phase for the subsequent attachment of a broad range of biochemically active molecules on the titanium surface. PMID- 15348880 TI - The healing process of palatal tissues after palatal surgery with and without implantation of membranes: an experimental study in dogs. AB - The aim of this study was to evaluate the wound-healing process clinically and histologically in growing beagle dogs after palatal repair according to von Langenbeck, with and without implantation of membranes of a copolymer of polyhydroxybutyrate 80%-hydroxyvalerate 20% (=PHB-co-HV 80/20). Von Langenbeck's repair was performed in 12 dogs (age 12 wk), while von Langenbeck's repair followed by implantation of PHV-co-HV membranes was carried out in 11 dogs (age 12 wk). Four dogs (age 12 wk) served as unoperated controls. Standardized intra oral slides of the palate were taken and measurements of the wound surface areas were carried out. Histological sections were prepared at three different ages. The animals were studied until the age of 25 wk. It was found that wound closure after the von Langenbeck's procedure took about 3 wk, while the use of PHB-co-HV membranes after von Langenbeck's repair resulted in complete wound closure after approximately 7 wk after the membranes had sequestered. At the age of 25 wk, the histologic results after the von Langenbeck procedure showed that the entire scar tissue covering the former denuded bony areas was attached to the bone by means of Sharpey's fibres, while after implantation of the membranes only local scar tissue attachment by means of Sharpey's fibres was found. Further research is necessary to develop a membrane which allows wound closure without sequestration of it. PMID- 15348881 TI - Electron paramagnetic resonance study of free-radical kinetics in ultraviolet light cured dimethacrylate copolymers. AB - A spin-trapping technique and direct kinetic measurements of electron paramagnetic resonance spectra were used to investigate the kinetics of radical production in the cross-linking photo-initiator system based on camphoroquinone and dimethyl-aminoethylmethacrylate in the presence of the corresponding monomer dimethacrylate copolymer mixture, as well as in the pure photo-initiator system. Consequently, in both cases, the kinetics of radical production and decay was measured. From the measured data the formal kinetic parameters of particular radical processes were calculated by the non-linear least-squares regression method. PMID- 15348882 TI - Mechanical properties and bone densities of canine trabecular bone. AB - The purposes of this study were (1) to evaluate the mechanical properties of canine epiphyseal cancellous bones from adult canine femoral heads, femoral condyles, tibial plateau, and humeral heads, using indentation and compression tests, and (2) to measure bone densities (apparent density and ash density) of these cancellous bones so as to develop a normal data base of mechanical strength and bone density. The correlations between the two mechanical tests and between these tests and bone densities were also considered. The results showed all of the three mechanical parameters, ultimate load, stiffness, and ultimate strength, measured by the indentation test were higher than those measured by the compression test. Correlation analysis showed that the two sets of mechanical values correlated well (r=0.823-0.952, p<0.01). The apparent density and ash density correlated well with the mechanical parameters determined by the two types of mechanical tests (r=0.737-0.966, p<0.05). PMID- 15348883 TI - Studies on the setting of polyelectrolyte cements: part VI. The effect of halide salts on the mechanical properties and water balance of zinc polycarboxylate and glass-ionomer dental cements. AB - A study is reported in which a zinc polycarboxylate and a glass polyalkenoate dental cement, respectively, were prepared from aqueous solutions of NaCl, KCl, KBr and KI, all at 1 mol dm3 concentration, as well as from pure water. For the zinc polycarboxylate, setting as determined by oscillating rheometry was speeded up and water uptake was enhanced by the presence of the salts. Conversely, compressive strength at 24 h was unaffected. On the other hand, for the glass polyalkenoate, the setting reaction was slowed down, water uptake inhibited and compressive strength at 24 h reduced (from 94.3 MPa with pure water to 59.8 MPa with NaCl, 65.8 MPa for KCl, 67.0 MPa for KBr and 81.1 MPa for KI). Previous work with polyelectrolytes in aqueous solution suggests that the halides probably enhance the rate of the neutralization process. For the zinc polycarboxylate, this leads to a more rapid setting reaction. By contrast, for the glass polyalkenoate, it results in slower setting and weaker cements. This result is attributed to inhibition of the secondary setting reaction, involving the formation of the silicate/phosphate network, by enhanced neutralization, a process which is consequently concluded to occur earlier in the overall setting of these cements than had been assumed previously. PMID- 15348884 TI - Studies in the setting of polyelectrolyte cements: part VII. The effect of divalent metal chlorides on the properties of zinc polycarboxylate and glass ionomer dental cements. AB - A study is reported in which a zinc polycarboxylate and a glass polyalkenoate dental cement, were prepared from aqueous solutions of divalent metal chlorides, namely ZnCl2, CaCl2, MgCl2 and SrCl2, all at 1.0 mol dm-3 concentration, as well as from pure water. Calcium chloride was employed at additional concentrations, i.e. 2.0, 0.5 and 0.1 mol dm-3. As was previously found for monovalent salts, setting of the zinc polycarboxylate was speeded up and water uptake generally enhanced by the presence of the divalent metal salts. However, the divalent salts were found to reduce the compressive strength at 24 h (from 86 MPa to about 60 MPa). The glass polyalkenoate showed broadly similar effects to those observed in the presence of monovalent salts, with the setting time being increased, water uptake inhibited and compressive strength at 24 h reduced; however, by contrast, the working time was generally reduced. These results occur because the rate of the neutralization process is increased by the divalent salts, a consequence of the reduced pH of the poly(acrylic acid) caused by these salts. Infrared spectroscopy demonstrated interactions between the metal chlorides and poly(acrylic acid), with various chelate structures being apparent from the position of the asymmetric carbonyl stretch. PMID- 15348885 TI - Apatite formation on silica gel in simulated body fluid: effects of structural modification with solvent-exchange. AB - The prerequisite for glasses and glass-ceramics to bond to living bone is the formation of biologically active bone-like apatite on their surfaces. It has been shown that even a pure silica gel forms the bone-like apatite on its surface in a simulated body fluid. In the present study, pore structure of silica gels prepared by hydrolysis and polycondensation of tetraethoxysilane in an aqueous solution containing polyethylene glycol was modified by 1M HNO3, and 0.1M and 1M NH4OH solution treatments. The three kinds of resultant gels all contained large amounts of silanol groups and trisiloxane rings, but differ greatly in pore structure of nanometre pore size. Irrespective of these differences, all the gels formed the bone-like apatite on their surface in the simulated body fluid. It was speculated that a certain type of structural unit of silanol groups, which is easily formed in the presence of the polyethylene glycol, is effective for the apatite formation. PMID- 15348886 TI - Apatite-organic polymer composites prepared by a biomimetic process: improvement in adhesion of the apatite layer to the substrate by ultraviolet irradiation. AB - A dense and uniform layer of highly bioactive apatite can be formed in arbitrary thickness on any kind and shape of organic polymer substrates by the following biomimetic process. The substrate is first placed in contact with granular particles of CaO, SiO2-based glass soaked in a simulated body fluid with ion concentrations nearly equal to those of human blood plasma for forming apatite nuclei, and then soaked in another fluid highly supersaturated with respect to the apatite for making the apatite nuclei grow. In the present study, the polymer substrates were pretreated with ultraviolet (UV) light, and then subjected to the biomimetic process described above. By UV irradiation, the induction period for the apatite nucleation of poly(ethylene terephthalate) (PET), poly-ether sulphone (PESF), polyethylene (PE), poly(methyl methacrylate) (PMMA) and polyamide 6 (N6) substrates were reduced form 24 h to 10 h. The adhesive strengths of the apatite layer to the substrates increased from 2.5-3.2 MPa to 4.5-6.0 MPa for PET, PESF and PMMA, and from about 1.0 MPa to 4.0-6.5 MPa for PE and N6 substrates. These results have been explained by assuming that silicate ions, which induce apatite nucleation, are easily adsorbed on the substrates due to the formation of polar groups, with an improved hydrophilic nature, on the polymer surfaces by UV irradiation. PMID- 15348887 TI - Evaluation of subsurface damage in CAD/CAM machined dental ceramics. AB - Commercial ceramics for dental computer aided design/computer aided manufacture (CAD/CAM) restorations suffer from surface chipping defects and microcracking. The influence of CAD/CAM machining of dental materials on the mechanical strength and extension of the damage zone was studied. Two different commercial dental ceramics, a feldspathic porcelain and a glass-ceramic, were CAD/CAM machined according to dental practice. The extension of the damage zone was analysed by a stepwise erosion of the surface, and the biaxial flexural strength was measured. To simulate the adhesive fixing of ceramic inlays, the specimens were sealed using a light-curing monomer. The different machining behaviour is dominated by the microstructure of the investigated materials. Owing to the high amount of glassy phase, the feldspathic porcelain shows extensive microcracking and chipping defects. The extent of the damage zone can be determined as 40-60 microm. Sealing of the surface did not affect the flexural strength of the machined samples. The dominating response to machining of the glass-ceramic is crushing and crumbling with a major contribution of plastic deformation on a microscopic scale. The extent of the damage zone is less than 20 microm. These cracks can be bridged by sealing of the surface, resulting in a substantial increase in strength. PMID- 15348888 TI - Calcium phosphate coating on titanium substrate by a modified electrocrystallization process. AB - Modification of ethyl alcohol added aqueous electrolyte for depositing calcium phosphate on titanium substrates by a electrocrystallization method is described. Film coated in the electrolyte with ethyl alcohol addition is more homogeneous and the growth rate is higher. The optimum quantity of ethyl alcohol added is 50% of the electrolyte. Although the pH value of electrolyte varies as ethyl alcohol is added, the phases of the deposited film remain the same, and are hydroxyapatite and brushite. The development of the microstructure of the coated film during deposition is discussed. PMID- 15348889 TI - Quantitative histological analysis of bony ingrowth within the biomaterial Polyactive implanted in different bone locations: an experimental study in rabbits. AB - The quantity of bone formed in cylinders of a newly developed erodible copolymer, Polyactive (PA60/40) was examined. PA60/40 was implanted in three different bone locations in the rabbit: in the cortex, in bone marrow and in trabecular subchondral bone. Bony ingrowth was assessed after 4, 8, 26 and 52 w after the operation and investigated by histology and image analysis. The ingrowth of bone was observed in PA60/40 placed in the cortex from 4 w onwards. After 8 w, more than 90% of the pores of the biomaterial were filled with dense bone. In bone marrow, initially some bone formation was seen. After 26 w, all newly formed bone was resorbed. Subchondral bone formation was less than in the cortex of the femur, but somewhat comparable to the amount of bone found in healthy trabecular bone. Bone formation appeared not to be affected by the degradation of the biomaterial. It was concluded that Polyactive is a suitable bone graft substitute. Bone formation within PA60/40 is site-dependent and this follows Wolff 's law. PMID- 15348890 TI - Nuclear magnetic resonance imaging - a potential method for analysis of bone material. AB - The basic usage of nuclear magnetic resonance (NMR) microimaging for materials characterization is introduced. An application of microimaging for three dimensional structural study of trabecular bone is presented with elementary types of image presentation. Solid bone in itself does not produce any significant NMR intensity and natural lipids and/or added 70% ethanol has been used as a signal source. The applied pulse sequences were three-dimensional spin echo and three-dimensional chemical shift selective spin echo. PMID- 15348891 TI - A finite-element model study of occlusal schemes in full-arch implant restoration. AB - A three-dimensional finite-element model of a human mandible is presented, and the stresses and deformations computed for loading states induced by two different gnathologic reconstructions using six and four implants are discussed. Occlusal canine guidance and posterior and anterior group functions on cantilevered and distally supported prostheses have been simulated. The stress distributions generated by the different loading conditions on either the osseointegrated prosthesis or the bone tissue surrounding the implants are described. The analysis of the stress distribution on the working side reveals that the posterior group function undergoes a reduction in stress intensity on the cortical bone surrounding the implants (especially for the distal implant) compared with the anterior group function and canine guidance in both gnathologic reconstructions. PMID- 15348892 TI - Hydrolysis of dicalcium phosphate dihydrate to hydroxyapatite. AB - Dicalcium phosphate dihydrate (DCPD) was hydrolysed in water and in 1 M Na2HPO4 solution at temperatures from 25-60 degrees C. Hydrolysis was incomplete in water. At 25 degrees C, DCPD partially hydrolysed to hydroxyapatite (HAp). Formation of HAp is indicative of incongruent DCPD dissolution. At the higher temperatures, hydrolysis to HAp was more extensive and was accompanied by the formation of anhydrous dicalcium phosphate (DCP). Both of these processes are endothermic. When hydrolysis was carried out in 1 M Na2HPO4 solution, heat absorption was greater at any given temperature than for hydrolysis in water. Complete hydrolysis to HAp occurred in this solution. The hydrolysis of DCPD to HAp in sodium phosphate solution was also endothermic. The complete conversion of DCPD to HAp in sodium phosphate solution would not be expected if the only effect of this solution was to cause DCPD dissolution to become congruent. Because of the buffering capacity of a dibasic sodium phosphate solution, DCPD hydrolysed completely to HAp. Complete conversion to HAp was accompanied by the conversion of dibasic sodium phosphate to monobasic sodium phosphate. The formation of DCP was not observed indicating that the sodium phosphate solution precluded the DCPD to-DCP dehydration reaction. In addition to affecting the extent of hydrolysis, reaction in the sodium phosphate solution also caused a morphological change in the HAp which formed. HAp formed by hydrolysis in water was needle-like to globular while that formed in the sodium phosphate solution exhibited a florette like morphology. PMID- 15348893 TI - Preparation of hydroxyapatite-granule-implanted superplastic titanium-alloy. AB - In order to obtain a biomaterial that has both biological affinity and high mechanical strength, hydroxyapatite (HAp) granules were implanted into the surface of superplastic titanium-alloy. HAp granules (32-38 microm diameter) were spread over a superplastic titanium-alloy substrate and pressed to implant the granules into the substrate. This was achieved at 17 MPa, 750 degrees C for 10 min. Only the tops of the granules, which were surrounded by the alloy, remained exposed and they were firmly stuck in the substrate. The granules were enclosed in titanium-alloy and a reaction layer was formed at their interface. The HAp implanted titanium-alloy composites are expected to be useful as biomaterials, such as artificial bones and dental roots. PMID- 15348894 TI - The influence of sterilization technique and ageing on the structure and morphology of medical-grade ultrahigh molecular weight polyethylene. AB - The effects of four sterilization treatments (gamma radiation in nitrogen, electron-beam radiation, ethylene oxide gas, and no sterilization) on the structure and morphology of ultrahigh molecular weight polyethylene (UHMWPE) were monitored as a function of ageing time in air for a period of 1.5 y. Characterization techniques employed include differential scanning calorimetry, density gradient column, transmission electron microscopy, and small-angle X-ray scattering. Ethylene oxide gas does not affect the structure of the polymer. Both forms of radiation lead to measurable alterations of the material's structure, including an increase in crystallinity, an increase in density, and the enhancement of lamellae crystalline stacking. Most changes in structure occur in the first few months with little differences observed upon subsequent ageing in air. The sharpness of the crystalline-amorphous boundaries decreases with time for irradiated UHMWPE and is believed to be linked to the oxidation of the polymer. PMID- 15348895 TI - Ageing behaviour in a dental low-gold alloy with high copper content. AB - Phase transformation behaviour in a dental low-gold alloy with high copper content during continuous heating was investigated by hardness tests, electrical resistivity measurements, X-ray diffraction, scanning and transmission electron microscopies. Two kinds of solution treatment conditions (at 873 K and 1073 K) followed by iced-brine quench, represented different ageing behaviours. Although subsequent anisothermal annealing produced same phase separation of face-centred cubic disordered and ordered (Cu3Au) phases in both specimens, the specimen quenched from 1073 K had already been hardened by a spinodal decomposition. PMID- 15348896 TI - Dynamic fatigue and degradation in solution of hydroxyapatite ceramics. AB - Polycrystalline hydroxyapatite was densified by hot pressing. The dissolution process in aqueous solution and the effects of environment on dynamic fatigue resistance of the resulting HAP ceramics were investigated. Pure water or Ringer's solution strongly enhances subcritical crack growth. The crack propagation exponent decreases from 22.5+/-2 in air to 10+/-4 in Ringer's solution for materials densified at 98% of the theoretical value. The residual porosity ratio is also very detrimental for the mechanical reliability. Both fatigue resistance and immediate fracture strength are decreased, with values of only 14+/-4 for the propagation exponent and 40 MPa for the tensile strength (compared to 90 MPa at 98% relative density) for materials densified at 94% tested in air. The degradation in solution appears to be governed by uniform physico-chemical dissolution of crystalline HAP phase at the surface of the material. This dissolution is accompanied by a decohesion of grains located around residual pores which leads to the growth of local surface defects. PMID- 15348897 TI - Chemical and structural changes in zinc polycarboxylate cements after immersion in dilute organic acid solutions. AB - The behaviour of zinc polycarboxylate cements in contact with dilute aqueous solutions of organic acids at concentrations close to those existing in buccal medium, was studied. The organic acids were acetic, citric, tartaric and lactic acids, at 0.01 M and 0.001 M. The elution of zinc and magnesium was 10-1000 times greater in acid than in pure water, and correlated with the concentrations and the dissociation constants, pK1, of the acids tested. In all cases, important water losses were observed. In the 0.01 M acids, the cement structure collapsed to form a viscous, compact and homogeneous layer on the cement surface. In this layer, the polymeric carboxylic chains were regenerated from the zinc and magnesium polycarboxylate cement. Comparison with pure water showed that even the smallest concentration of the weak acids greatly modified the cement behaviour. This could explain the well-known differences in erosion processes between theoretical erosion predicted by standard specification tests and the in vivo situation. PMID- 15348898 TI - In vivo cytogenetic studies of the genotoxic effects of polymethyl methacrylate employed in orthopaedics. AB - Polymethyl methacrylate (PMMA) is a material employed as a cement in prosthesis that has been found to induce cytogenetic damage in human lymphocytes in vitro. This effect is particularly achieved before the 5th day of the exposition, while after this period no significant variations appeared. In the present study, results of cytogenetic tests in humans are reported. Sister chromatid exchange analysis and micronucleus tests have been performed on lymphocytes of patients who had undergone prosthesization with cemented prosthesis (group A) or with biological anchorage prosthesis, as control (group B). DNA damage was investigated before implantation and 5 d after surgery in both groups. Cytogenetic tests did not show any significant increase in the number of micronuclei and sister chromatid exchanges with respect to control values in patients with PMMA cemented prosthesis. PMID- 15348899 TI - Behaviour of photopolymerized silicate-glass-fibre-reinforced dimethacrylate composites subjected to hydrothermal ageing: part I. Steady-state sorption characteristics. AB - Steady-state water sorption due to hydrothermal ageing (37 degrees C) was determined via gravimetric methods for both an unreinforced dimethacrylate copolymer network and for composites of the copolymer reinforced with two different silicate yarn products - S2-glass(R) and quartz. The copolymer sorbed 4.9+/-0.3 wt%, and the composites sorbed 0.5-2.5 wt%. For the two types of composites, two different positive linear correlations were established between the sorption gain and the copolymer content in the composites. In the case of the S2-glass(R) composites, the sorption gain was fully explained in terms of sorption in the copolymer; and normalization of these data by the mass fraction of copolymer yielded a constant sorption value matching that of the unreinforced copolymer. After normalizing the data for the quartz composites, an additional positive correlation was established between the water gain (per unit weight of copolymer) and the linear density of interfacial surface area. For both types of composites the sorption response was stable over a three-month ageing period. PMID- 15348900 TI - Preparation of bioactive microporous titanium surface by a new two-step chemical treatment. AB - Microporous oxide layers allowing fast deposition of calcium phosphate layers (CPLs) were formed on commercially pure titanium (c.p.Ti) after the application of a newly developed two-step chemical treatment. The micropores were of submicrometre size. The two-step treatment was carried out by etching c.p.Ti samples with HCl and H2SO4 first and then treating them in boiling 0.2 N NaOH solution at 140 degrees C for 5 h. Conformal CPLs, about 20 microm thick, were deposited on the two-step treated c.p.Ti surface by means of a two-day immersion in an in vitro supersaturated calcification solution. The CPL was characterized to be mainly composed of two sublayers, i.e. an outside loose octacalcium phosphate crystal sublayer and an inside dense carbonated apatite sublayer. A scratching test indicated that the apatite sublayer was strongly bonded to the c.p.Ti substrate. Moreover, it was observed that the untreated or single-step treated c.p.Ti surfaces are not only morphologically different from one another but significantly different from the two-step treated one, in that no precipitation was observed on them up to 14 d immersion in the same calcification solution. It is indicated that the two-step chemical treatment is a simple and easily controllable method to prepare bioactive titanium surfaces and subsequently to induce the rapid precipitation of conformal and adherent CPL from in vitro supersaturated calcification solutions. PMID- 15348901 TI - Antimicrobial effects of metal ions (Ag+, Cu2+, Zn2+) in hydroxyapatite. AB - The antimicrobial ceramics (AC) based on hydroxyapatite (HA) were made in a wet chemical process with additions of AgNO3, Cu(NO3)2. 3H2O and Zn(NO3)2. 6H2O. The ACs were composed of metal-ion substituted hydroxyapatite and nitrate-apatite, which was identified by X-ray diffraction. The viable count and turbidity measurement was adopted to observe the antimicrobial effects of the various ACs. The aerobic Escherichia coli was used in the study. An obvious antimicrobial effect against E. coli was observed in Ag+ AC. In contrast to Ag+ AC, it was difficult to ascertain any bactericidal effect in the case of Cu2+ and Zn2+ AC. The bactericidal effect of Ag+ was observed using a dialysis tube experiment. This suggests that Ag+ dissolved out and reacted with E. coli, thus inhibiting its growth. PMID- 15348902 TI - Adsorption of serum alpha-1-microglobulin onto biomaterials. AB - The adsorption of alpha-1-microglobulin (alpha-1-m) from serum to the surface of polymers with different physicochemical properties was investigated. Enzyme linked immunosorbent assay showed binding of this protein to the surface of polystyrene (PS), polyvinyl chloride (PVC) and a polyurethane, Chronoflex, after water washing, but only trace levels could be detected on two polymethacrylate derivatives, polymethyl methacrylate and poly(2-hydroxyethyl methacrylate). alpha 1-m was selectively desorbed from the five materials by sequential washes of serum-conditioned surfaces with isopropanol solutions at increasing concentrations. The presence of alpha-1-m in the washing supernatants was detected by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE). The relative binding strength of alpha-1-m to each surface was evaluated as the isopropanol (IsoPOH) concentration required to desorb the protein from that surface. Analysis of bound proteins by SDS-PAGE conclusively demonstrated the binding of a range of serum proteins, including alpha-1-m, to all polymer systems, but with varying binding strengths. The majority of protein was removed by water washing for the polymethacrylate polymers, while varying concentrations of IsoPOH were required to desorb proteins from PS, PVC and Chronoflex. There was a correlation between the hydrophobic nature of the material, determined by water contact angle measurements, and adsorption of alpha-1-m. Immunoblotting of isopropanol-eluted proteins by alpha-1-m antibodies showed the positive staining of a 29 kDa protein as well as selected bands within a molecular weight range of 40 200 kDa, suggesting the adsorption of this protein as both free and complexed forms. The ability of alpha-1-m to adsorb on to material surfaces and to participate in events relevant to the biocompatibility of a polymer, such as bacterial infection or inflammation control, suggests the need for further characterization of the properties of this protein. PMID- 15348903 TI - Detection of mutagenic potential of some glass-ionomer cements through Ames testing. AB - The mutagenic potential of three commercially available glass-ionomer cements used in dentistry was examined. The cement components were mixed according to the manufacturers indications and set for two defined times: 1 h or, alternatively, 1 wk. Cements B and C set spontaneously; in the case of cement A, the manufacturer suggests the use of a lamp to trigger also a photopolymerization. Photopolymerization, however, was not used. Ames tests were performed on the dimethyl sulphoxide extracts of cements by using Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, TA 1538 and TA 102. Cement A showed mutagenicity only against TA 1537 strain, either in the presence or absence of metabolic activation with microsomial fraction S9. The other two cements showed no mutagenic potential. We conclude that glass-ionomer cements are, on the whole, safe materials from the viewpoint of genotoxicity, and hypothesize that the mutagenicity observed in cement A could depend on its polymerization performed without light activation. PMID- 15348904 TI - In vitro and in vivo microbial adhesion and growth on argon plasma-treated silicone rubber voice prostheses. AB - Patients who undergo a total laryngectomy usually receive a silicone rubber voice prosthesis for voice rehabilitation. Unfortunately, biofilm formation on the esophageal side of voice prostheses limits their lifetime to 3-4 mon on average. The effects of repeated argon plasma treatment of medical grade, hydrophobic silicone rubber on in vitro adhesion and growth of bacteria and yeasts isolated from voice prostheses, as well as in vivo biofilm formation are presented here. In vitro experiments demonstrated that initial microbial adhesion over a 4 h time span to plasma-treated, hydrophilized, silicone rubber was generally less than on original, hydrophobic silicone rubber, both in the absence and presence of a salivary conditioning film on the biomaterial. Growth studies over a time period of 14 d at 37 degrees C in a modified Robbins device, showed that fewer Candida cells adhered on plasma-treated, hydrophilized silicone rubber as compared to on original, hydrophobic silicone rubber. For the in vivo evaluation of biofilm formation on plasma-treated silicone rubber voice prostheses, seven laryngectomized patients received a partly hydrophilized "Groningen Button" voice prosthesis for a planned evaluation period of 4 wk. After removal of the voice prostheses, the border between the hydrophilized and the original, hydrophobic side of the prostheses was clearly visible. However, biofilm formation was, unexpectedly, less on the original, hydrophobic sides, although the microbial compositions of the biofilms on both sides were not significantly different. Summarizing, this study demonstrates that in vitro microbial adhesion and growth on silicone rubber can be reduced by plasma treatment, but in vivo biofilm formation on silicone rubber voice prostheses is oppositely enhanced by hydrophilizing the silicone rubber surface. Nevertheless, from the results of this study the important conclusion can be drawn that in vivo biofilm formation on voice prostheses is controlled by the hydrophobicity of the biomaterials surface used. PMID- 15348905 TI - Sol-gel derived porous hydroxyapatite coatings. AB - The preparation of hydroxyapatite coatings on alumina substrates by a sol-gel method was investigated. A mixed ethanol solution of Ca(NO3)2.4H2O and P2O5 was used as a dipping source. The samples were characterized by solid-state 31P nuclear magnetic resonance spectroscopy, differential thermal analysis, thermogravimetry, X-ray diffraction, scanning electron microscopy and infrared reflection spectroscopy. The coating obtained at 500 degrees C had good hydroxyapatite crystallinity, adhesive strength of about 10 MPa and dense morphology. The coating obtained at 750 degrees C also had good crystallinity and adhesive strength but porous morphology. PMID- 15348906 TI - Analysis of the effect of pressure on compression moulding of UHMWPE. AB - Ultra-high molecular weight polyethylene (UHMWPE) powder is effectively processed by compression moulding due to its very high melt viscosity. Compression moulding involves application of temperature and pressure as a function of time. The pressure applied during processing has a significant influence on the part properties. The effect of pressure applied during compression moulding was studied by moulding parts at different pressures. Increase in the applied pressure causes increase in the melting and recrystallization temperatures. An increase in the pressure applied at the melt temperature ( approximately 140 degrees C) from 7.8 MPa to 15.6 MPa caused the crystallinity to increase from 54% to 61%, the stiffness of the moulded part to increase from 257 MPa to 435 MPa and oxidative index to increase from 0.055 to 0.059. Further increase in the pressure applied at the melt to 23 MPa caused the crystallinity to fall to 49%, the modulus to reduce significantly to 302 MPa and the oxidative index to change to 0.063. Increase in the pressure applied at the recrystallization temperature ( approximately 91 degrees C) from 38 MPa to 78 MPa increased the crystallinity from 54% to 65%, increased the modulus from 257 MPa to 279 MPa and increased the oxidative index from 0.055 to 0.065. Further increase in the applied pressure to 97 MPa, caused the crystallinity to drop slightly to 61% the modulus to reduce to 269 MPa, and the oxidative index to reduce to 0.057. The experiments showed that for obtaining maximum crystallinity and stiffness, the applied pressure should be within a narrow range. The highest recrystallization pressure (97 MPa) indicated the formation of extended-chain crystals in addition to the chain-folded crystals. The change in pressure applied at the melt temperature had a significantly greater effect on Young's modulus, as compared to change in pressure applied at the recrystallization temperature. Fourier transform-infrared spectroscopy analysis of the samples moulded at different pressures revealed that the increase in crystallinity and stiffness was accompanied by increase in oxidation within the part. By filling the die in a nitrogen atmosphere instead of air, the oxidation level in the moulded parts was reduced by almost 60%, without adversely affecting the crystallinity and the modulus. PMID- 15348907 TI - Resorbable continuous-fibre reinforced polymers for osteosynthesis. AB - Four institutes from three countries in the European Union have collaborated under the BRITE-EURAM framework programme for the development of processing technologies for resorbable osteosynthesis devices. The devices should be continuous-fibre reinforced, and the technology should offer the possibility of orienting the fibres in the main trajectories. Poly-L-lactide and poly-L-DL lactides have been synthesized for reinforcement fibres and matrix material, respectively. Melt-spun P-L-LA fibres of a strength of 800 MPa have been embedded in an amorphous P-L-DL-LA 70 : 30 matrix by compression moulding. Ethyleneoxide sterilized samples have been tested in vitro and in vivo. A satisfying bending modulus has been reached (6 GPa). Yet with 50% strength retention after ten weeks, fast degradation occurred that could be related to residual monomers. By this fast degradation 70% resorption after one year could be observed in the non functional animal studies in rabbits. There was only a mild inflammatory reaction, which confirmed the good biocompatibility of the materials even during the resorption period. Further effort has to concentrate on the reduction of initial monomer content. The great advantage of the processing method to orient fibres in the device will be utilized in prototype samples, e.g. an osteosynthesis plate with fixation holes. PMID- 15348908 TI - Chemical surface treatment of silicone for inducing its bioactivity. AB - It has been confirmed that the apatite nucleation is induced by silanol (Si-OH) groups formed on the surfaces of materials and/or silicate ions adsorbed on them. It was previously shown that apatite nuclei are formed on organic polymers when the polymers are placed on CaO, SiO2-based glass particles soaked in a simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma, and that they grow spontaneously to form a dense and uniform apatite layer together with high adhesive strength to the substrates when the polymers are soaked in another solution with ion concentrations 1.5 times the SBF. In the present study, silanol groups bonded covalently to the surface of the silicone substrate were formed and its apatite-forming ability was examined. When silicone substrates were treated with 5 or 10 M NaOH with pH 7.25 at 36.5 degrees C for more than 3 h, silanol groups were formed on the surfaces of the substrates. When thus NaOH-treated substrates were soaked in 1.5SBF at 36.5 degrees C, a bone-like apatite was formed on the substrates in a short period. PMID- 15348909 TI - Direct formation of nanophase hydroxyapatite on cathodically polarized electrodes. AB - Ultrafine-grained, nanophase coatings of hydroxyapatite were synthesized by electrocrystallization from dilute electrolytes ([Ca]=6.1 x 10-4 M, [phosphate]=3.6 x 10-4 M) at pH values comparable with the biological pH. At these comparatively low supersaturations, hydroxyapatite is shown to be precipitated without the formation of a precursor phase. A description of the sequence of events occurring at the electrode-electrolyte interface is given to explain the mechanism involved in the direct formation of nanophase hydroxyapatite on polarized electrodes. PMID- 15348910 TI - In vitro cytotoxicity testing of chitosan-containing polyelectrolyte complexes. AB - Two chitosan-containing polyelectrolyte complexes, chitosan-poly(acrylic acid) and chitosan-poly(styrenesulphonate), were synthesized by polymerizing acrylic acid and sodium styrenesulphonate in the presence of chitosan and chitosan hydrochloride, respectively. The complexes were studied by optical microscopy and tested for cytotoxicity by the Neutral Red uptake, Kenacid Blue R-Binding and 3 (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assays. The optical microscopy confirmed the differences in crystallinity and structure already found for the two polycomplexes by other characterization techniques. The cytoxicity tests showed different influences on the cell activity by the extracts of the two polyelectrolyte complexes. Such results were discussed and correlated to the different structures of the two materials. PMID- 15348911 TI - Analysis of shearing stress in the limited durability of bovine pericardium used as a biomaterial. AB - The objective of the study was to determine the shearing stress exerted by the suture thread under conditions of normal working stress. Thirty-six samples of calf pericardium, similar to that employed in the manufacture of bioprosthetic cardiac valve leaflets, were subjected to tensile testing. Prior to the trial, a continuous suture was sewn in the central zone of each sample, at a 45 degrees angle to the longest axis of the sample, using commercially-available threads (silk, Gore-Tex, Surgilene and nylon). Application of the Mohr circle for combined wear revealed that the shearing stress ranged between 2.68-fold greater (for samples sewn with silk) and 5.48-fold greater (for samples sewn with nylon) than the working tensile stress in the region of the suture. It is concluded that the shearing stress is responsible for the limited durability of sutured samples of calf pericardium prepared to simulate bioprosthetic cardiac valve leaflets. PMID- 15348912 TI - Adjustable forming of thermoplastic composites for orthopaedic applications. AB - The present study was focused on the development of a special thermoforming technique for manufacturing of continuous fibre reinforced thermoplastic composite parts with complex surface contours. In particular, a stamp forming process was modified to investigate the potential manufacturing advantages of thermoplastic composites in orthopaedic applications. An apparatus was designed which allowed the thermoforming procedure to be fully automatic, i.e. a cold pre consolidated laminate panel, as the feed material, was heated up in an infrared heating zone and then transferred into a cold mould system, where it was stamp formed. Both halves of the mould were made of many tiny round metal sticks in a metal frame. This needle-bed mould allowed one to copy any contour by pushing it slightly on spring fixed sticks. The desired position of these sticks could then be adjusted by forcing the side plates of the metal frame together. To prevent any press mark of stick-tops on the composite, i.e. to achieve smooth surfaces of the themoformed composite parts, flexible rubber pads were needed to cover the mould surfaces. Experimental results showed that the surface profile of CF/PP and GF/PP composites formed by the needle-bed mould reproduced fairly well the contour of a saddle shaped, complex model sample. Unique properties of this needle-bed mould are that it can be repeatedly used, and that it can copy any complex surface contours, for example a bone surface, by simply adjusting the stick positions according to the special surface requirements. PMID- 15348913 TI - Amount of hydroxyl radical on calcium-ion-implanted titanium and point of zero charge of constituent oxide of the surface-modified layer. AB - To compare the surface properties of calcium-ion (Ca2+)-implanted titanium with those of titanium and to investigate the mechanism of bone conductivity of Ca2+ implanted titanium, amounts of hydroxyl radical of Ca2+-implanted titanium and titanium were estimated. Also, the point of zero charge (p.z.c.) of oxide constituting surface oxides of Ca2+-implanted titanium and titanium was determined. Results showed that the amount of active hydroxyl radical on Ca2+ implanted titanium was found to be significantly larger than that on titanium, indicating that the number of electric-charging sites of Ca2+-implanted titanium in electrolyte is more than that of titanium. The p.z.c. values of rutile (TiO2), anatase (TiO2), and perovskite (CaTiO3), were estimated to be 4.6, 5.9, and 8.1, respectively. Thus, Ca2+-implanted titanium surface is charged more positively in bioliquid than titanium, accelerating the adsorption of phosphate ions. PMID- 15348914 TI - Preliminary histological study of connective tissue response to Zinalco and stainless steel 316L implants after 120 days. AB - Circular plates of Zinalco alloy (80 wt% Zn, 1.5 wt% Cu, 18.5 wt% Al) and stainless steel (SS) 316L were implanted in 12 female Wistar rats subcutaneously and intramuscularly to compare organism response, 120 days after implantation. The tissues surrounding the implants were analysed employing hematoxilin and eosin (H-E) and Gallego's trichromic techniques (GTT). Findings indicate that the reaction to Zinalco alloy was similar to the reaction to SS 316L. The Zn, Al and Cu concentrations in blood were measured, without evidence of any alteration due to implants. The presence and distribution of Zn, Al and Cu components of Zinalco alloy were detected in tissues by energy dispersive X-ray microanalysis. PMID- 15348915 TI - Effect of microstructure of titanium surface on the behaviour of osteogenic cell line MC3T3-E1. AB - Three different microstructures were obtained on a titanium surface via immersion in HCl, H3PO4, or mixed acid of HNO3 and HF (HNO3/HF) solution. The microstructure and Rmax of the acid-treated surfaces were dependent on the acid type and immersion conditions. The growth rate of the osteogenic cell line MC3T3 E1 on each acid-treated sample, which was measured using MTT-formazan assay, was significantly higher than that of the standard which was ground with #400 SiC grit paper. Moreover, both the H3PO4 treated sample and the HNO3/HF-treated surface showed a tendency to enhance the alkaline phosphatase activity of MC3T3 E1 cells, which were grown on each acid-treated surface. These results suggest that the acid treatment of titanium is effective for the improvement of its osteocompatibility. PMID- 15348916 TI - Hardness and fracture toughness of dense calcium-phosphate-based materials. AB - Hardness, H, and fracture toughness, KIc, have been determined as well as fracture energy and embrittlement index, H/KIc for six calcium-phosphate ceramics differing in phase composition. These materials were produced from initial calcium-phosphate precipitates with Ca-P molar ratios ranging between 1.50 and 1.73, synthesized by wet methods. After uniaxial or isostatic moulding and thermal treatment at 1250 degrees C, the obtained dense sinters constituted mono , bi- or triphase ceramic materials containing hydroxyapatite (HAp), beta-TCP, alpha-TCP and CaO. When comparing the investigated materials, the best parameters, i.e. relatively high hardness accompanied by high KIc, were observed in the case of a HAp-TCP composite, containing approximately 15 wt% HAp. It has been stated that free CaO occurring on the surface of the HAp samples obtained from powders with Ca-P ratios exceeding 1.67, transforms partially to CaCO3 due to contact with the surrounding atmosphere. The well shaped calcite crystals existing on the surfaces of such sinters significantly reduce hardness and increase fracture energy of the material when comparing both with the monophase HAp and the biphase HAp-TCP ceramics. PMID- 15348917 TI - X-ray diffraction of bone at the interface with hydroxyapatite-coated versus uncoated metal implants. AB - The microstructural characteristics of the newly formed bone tissue at the interface with hydroxyapatite-coated and uncoated stainless steel pins used in an external fracture fixation system have been evaluated. The bone far from the interface was used as a control. Pins were transversally inserted into the diaphyses of sheep tibiae and were loaded in for six weeks. Three sheep received coated pins and two received uncoated pins. Crystallographic habit and mineralization of the implant-facing bone were evaluated. Moreover, lattice parameters of bone apatite were measured and hydroxyapatite (HA) coating degradation was investigated, by means of conventional and microbeam X-ray diffraction (XRD). In coated pins, six weeks after the implantation the newly formed bone tissue at the interface did not reach complete maturation, but the presence of the implant did not alter the apatite lattice structure; the lattice parameters did not show statistically significant variations with respect to those observed in the control bone. In uncoated pins, bone tissue rarely appeared totally mineralized and lattice parameters were significantly different with respect to those observed in the bone far from the implant. HA particles were observed spreading in the bone-facing coated pins; the XRD pattern of bone apatite surrounding HA particles was unmodified. It was concluded that HA coatings improved the bone remodelling process during pin fixation in comparison to uncoated pins and did not alter the crystallographic habit of apatite. PMID- 15348919 TI - Homo sapiens has suddenly been confronted with a revolution. PMID- 15348918 TI - Adherent bacteria cells in five dental materials: sonication effect. AB - Adherent bacterial cells on the surfaces of two dental porcelain ceramics, three composite resins and human enamel were examined using four types of bacteria strains. Their adherent cells were counted on saliva-coated and uncoated material surfaces after sonication, and contact angle and zeta potential were measured for each adherent cell tested. A correlation between contact angle and bacterial cells on an uncoated surface was found to be higher in two Streptococcus sanguis cells than in S. mutans Ingbritt and S. sobrinus OMZ 176, whereas there appeared to be a higher correlation between S. mutans Ingbritt or S. sobrinus OMZ 176 and zeta potential on the uncoated surface. On the saliva-coated surface, a significantly high correlation was found between the adherent cells, with the exception of S. sanguis ATCC 10 557, and the zeta potential. Contact angle and zeta potential values were small when the surfaces of the materials were coated with saliva, as compared with those on the uncoated surface. The sonication condition (120 s) of adherent cells on the surface of the material significantly depended on the types of bacteria cells, showing that S. mutans Ingbritt (>50 60%) had a greater removal percentage than the others (<50%). PMID- 15348920 TI - A new biomedical polyurethane with a high modulus based on 1,4-butanediisocyanate and epsilon-caprolactone. AB - A new approach to the synthesis of biomedical polyurethanes based on epsilon caprolactone and 1,4-butanediisocyanate with a high modulus, has been developed. By chain extending an epsilon-caprolactone prepolymer with a long uniform-size diisocyanate block, a segmented polyurethane with uniform-size hard segments was obtained. It shows excellent mechanical properties; an extremely high modulus of 105 MPa and a tensile strength of 35 MPa. The polymer is soluble at high concentrations in various volatile solvents such as chloroform and 1,4-dioxane. By a combination of salt-leaching and freeze-drying, porous materials have been obtained in which macropores ranging in size from 150-300 microm are highly interconnected by micropores. The material shows a sufficiently high compression modulus of 200 kPa and appears to be suitable for biomedical applications such as meniscal prostheses. PMID- 15348921 TI - Acrylic bone cements incorporating polymeric active components derived from salicylic acid: curing parameters and properties. AB - A methacrylic monomer derived from salicylic acid, 5-hydroxy-2-methacrylamido benzoic acid, 5-HMA, was incorporated with 2-hydroxyethyl methacrylate, (HEMA), in different proportions to the liquid phase of classical bone cement formulations. The monomer 5-HMA shows the ability to form molecular complexes with calcium atoms in order to improve osteointegration in the application of bone cement formulations used for the fixation of joint prostheses such as knee and hip. Kinetic parameters, peak temperature and setting time of the bone cement formulations prepared were determined, obtaining lower peak temperature values when 5-HMA was incorporated, with respect to classical acrylic bone cements based on PMMA. Mechanical and thermal properties as well as surface energy values, have been determined for all cured bone cement formulations. PMID- 15348922 TI - Friction in hip-joint prostheses and its influence on the fixation of the artificial head. AB - The head of an implanted hip joint endoprosthesis is exposed to torques, which are transferred during gait due to the friction between the head and the cup prosthesis. In prostheses with ceramic ball heads, which are widely used now, and in which the head is fixed onto the stem by conical clamping, these torques could possibly affect the connection. In this study, torques transferred from the cup to the head are compared to the torques which are required to loosen the head from the metallic spigot. The results show that for the investigated head and taper types and sizes, under normal conditions the connection is safe with respect to undesired rotation. However, it is shown that for polluted sliding surfaces the fixation strength could possibly be exceeded. PMID- 15348923 TI - Quantification of the bone-related mRNAs at the bone/prosthetic interface. AB - Aseptic loosening is one of the major reasons for failure of joint prostheses. The periprosthetic tissue has previously been described microscopically; however, little work has been devoted towards quantitating genes expressed by cells at the materials/tissue interface. This study aims to characterize the phenotypic expression of osteoblasts and test the feasibility of quantifying the level of gene expression in periprosthetic tissue sections by combining in situ hybridization and image analysis techniques. There are many factors to consider when quantifying mRNA, in that comparing labeling between different cDNA probes, these should have comparable length and base comparison. The probes should be labeled with the same specific activity, that is the amount of probe to label added is the same, both between different probes and between batches of the same probe. Chromagen color reactions are variable in that the color development is not always linear and more likely follows a sigmoidal curve. Samples should only be compared when it is known that the reaction has been in the linear range. The image analysis of such staining introduces further factors which should be considered and controlled. Color analysis is a very complex problem with respect to reproducibly analyzing histological sections. The brightness component of the image should be independent of the colors within the image, in conventional RGB (red, green and blue) signalling mode this is not possible, while when using HSI (hue saturation and intensity) mode this becomes possible, and factors like staining intensity and brightness of the image become much more accountable and controllable. With these factors identified, we consider that the quantitative image analysis approach does allow comparison of patterns of bone-related mRNAs and demonstrates differences in expression in these osteogenic factors depending on distance from the prosthesis, tissue type, patient and device. PMID- 15348924 TI - Persistent expression of mitogenic/transforming factors at the site of failed orthopaedic implants: the impact on immune reactivity. AB - The response to wear particles from orthopaedic implants can lead to inflammation, osteolytic lesions, and aseptic loosening. To gain an insight into the development of this pathogenetic process, immunohistochemical techniques were used to identify the expression and tissue distribution of the potent cell mitogen epidermal growth factor (EGF), and the epidermal growth factor receptor (EGF-R) at the site of bone erosion in 30 patients with clinically failed orthopaedic implants. The results showed a large proportion of the macrophage subsets (Mphi) which expressed EGF and EGF-R, also contained wear particles, indicating their expression is a consequence of Mphi phagocytosis of implant material. The surface membrane expression of EGF-R on fusing Mphi suggests its presence is fundamental to the formation of bone-resorbing multi-nucleated giant cells, and the development of osteolysis. Additionally, there is increasing evidence of the long-term systemic spread of wear particles and their accumulation at distal sites including lymph nodes, liver, and spleen. Elevated expression of mitogenic factors in response to wear particles may result in deviation from normal cell growth and regulation, resulting in changes to immune cell function. Such potential transformations at distal sites are clinically significant, as alterations to the patient's immune system may result in acute divergence from normal immune cell responses. PMID- 15348925 TI - In vivo skeletal response and biomechanical assessment of two novel polyalkenoate cements following femoral implantation in the female New Zealand White rabbit. AB - Glass-ionomer cements (GIC) offer several advantages over the conventional acrylic-based bone cements. The formation of an adhesive bond with bone and metals, a low setting exotherm and no systemic or local toxicity are some of the advantages cited. This study examines the in vivo biological and biomechanical behavior of two polyalkenoate cements (LG26 and LG30) implanted for 6 wk into the submetaphyseal spongiosa of the rabbit femur. Cements were implanted as both set cement rods and unset cement dough. Implantation of set rods resulted in the formation of variably mineralized osteoid/woven bone at the bone-cement interface. Mechanical (push-out) testing revealed the strength of this bone cement interface was of similar magnitude to control (PMMA-rod implanted) animals. The bone of LG cement-dough implanted animals exhibited demineralization of pre-existing bone local to the site of implantation, accumulation of aluminum both locally and at a distance from the site of implantation, and defective mineralization of newly formed osteoid. The histological picture following LG implantation was strikingly similar to human renal osteodystrophy, in which skeletal accumulation of aluminum is a noted feature. The development of a GIC with low/no aluminum release from the unset cement dough is a priority in the further development of these cements for possible orthopaedic applications. PMID- 15348926 TI - Fatigue of bone cement with simulated stem interface porosity. AB - Cracks in bone cement have been observed in carefully examined post-mortem preparations of cemented stems. These cracks were probably caused by fatigue, and frequently appeared to initiate at pores. Ubiquitous porosity, occurring preferentially at, or near, the stem, is most likely caused by polymerization shrinkage. Preparation of air-free cement has only a marginal influence on the interface porosity, but pre-heating the stem in order to reverse the direction of polymerization can reduce or eliminate it. To estimate the impact of interface porosity on the fatigue strength of bone cement, test plates for this study were cast in a steel mold without release foils, and with one side of the mold warmer. Sample plates so prepared from chilled, partial vacuum-mixed PALACOS, have one face essentially pore-free and the other porous, the extent and morphology of the porosity being very similar to that observed on the stem-cement interface. Four point bending fatigue strength, determined after 60 d conditioning in Ringer's solution at 37 degrees C, was only 20 MPa (at 10(6) cycles, with the porous side under tension) compared to 30 MPa for conventionally prepared, pore-free material. This corresponds to a 10-100 fold reduction in cycles to failure in the range of stresses predicted to occur in vivo. PMID- 15348927 TI - Functionality of MDCK kidney tubular cells on flat polymer membranes for biohybrid kidney. AB - The prerequisite for the development of a biohybrid artificial kidney, is a substrate for confluent growth of renal cells forming an epithelial monolayer without any leaks. Conventional cell culture supports cannot be adapted for this purpose, because they lack adequate mechanical properties and thermal stability. From two suitable materials, polysulfone and polyacrylonitrile, two permeable polymeric membranes have been produced that were, according to ISO 10993-5, not cytotoxic. Cloned Madin Darby Canine Kidney (MDCK) cells (an established renal cell line) were cultured on the surface of the plastic materials, and on conventional cell culture supports. With all materials, assays of mitochondrial and lactate dyhydrogenases exhibited similar proliferation and the viability of the MDCK cells. Transmission electron microscopy showed the expression of a normal morphology of kidney tubular cells. Perfect barrier function, consequent on the formation of intercellular junctions in a confluent tight epithelium, was visualized in electron micrographs, and quantified by measurement of the transepithelial resistance. The uniformity of the cells grown was demonstrated in samples by electron microscopy and in the whole epithelium by intravital impedance analysis. It was concluded that polymeric membranes produced from polysulfone or polyacrylonitrile are appropriate substrates in the design of biohybrid kidney devices. PMID- 15348928 TI - Bone morphogenetic protein and ceramic-induced osteogenesis. AB - To investigate the relationship between bone morphogenetic proteins (BMP) and calcium phosphate ceramic-induced osteogenesis in soft tissues, in vitro and in vivo experiments were performed. In an in vitro study, the ability of different calcium phosphate ceramics to absorb bovine BMP (bBMP) from a bBMP solution was tested. In vivo studies included immunohistochemical BMP staining before bone formation in the ceramics was detected, and the enhancement of bone formation in calcium phosphate ceramics by bBMP. The results were: (1) calcium phosphate ceramics have a strong ability to absorb bBMP; (2) a high BMP concentration reaches inside the ceramic implants before bone formation in soft tissues of domestic pig occurs; (3) by 56% at 50 d and by 23% at 100 d, bBMP enhances bone formation in calcium phosphate ceramics implanted in soft tissues of dogs. The results indicate the BMP plays an important role in calcium phosphate ceramic induced osteogenesis and that adsorption of native BMP from the body fluids to ceramic implants may be a key step in osteoinduction by calcium phosphate ceramics. PMID- 15348929 TI - Osteoinduction by calcium phosphate biomaterials. AB - Different materials were implanted in muscles of dogs to study the osteoinduction of calcium phosphate biomaterials. Bone formation was only seen in calcium phosphate biomaterials with micropores, and could be found in hydroxyapatite (HA) ceramic, tricalcium phosphate/hydroxyapatite ceramic (BCP), beta-TCP ceramic and calcium phosphate cement. The osteoinductive potential was different in different materials. The results indicate that osteoinduction can be a property of calcium phosphate biomaterials when they exhibit specific chemical and structural characteristics. PMID- 15348930 TI - Interleukin 15 production by macrophages in the implant interface membrane of aseptically loosened joint replacements. AB - The T lymphocytes are cells involved in immunologically mediated hypersensitivity reactions. They are found in the cellular infiltrate present in the interface membrane of aseptically loosened joint prostheses. Activation of these cells has been demonstrated but not the important cytokine, interleukin 2 (IL2) required for such activation. This study describes the localization of IL15 and its mRNA in macrophages in interface membranes associated with activated T lymphocytes which are proliferating (as is evident from HLA-DR and Ki67 expression, respectively). The findings provide further evidence for immune mediated processes in aseptic loosening of orthopaedic implants because IL15 has similar activities to IL2 in T lymphocyte activation. PMID- 15348931 TI - Evaluation of the osteoblast response to a silica gel in vitro. AB - Many bioactive glasses and glass ceramics contain silica, yet the effect of silica on the osteoblast is not well understood. The osteoblast cell response to a silica surface, without the interference of the other ions present in glasses and glass ceramics has been investigated. A silica sol-gel was prepared which gave a molar ratio of 1:4:4 tetraethyl orthosilicate (TEOS): ethanol:acidified water 0.2 M HCl) and spin cast on to thermanox discs. The gel was characterized in terms of bioactivity and release of silicic acid. Primary human osteoblasts (HOBs) were seeded on the surface of upright or inverted silica discs. Cell activity (alamar blue reduction), number (DNA content) and differentiation (alkaline phosphatase activity, nodule formation and mineralization) were measured. There was no apparant difference in cell number, activity or alkaline phosphatase activity between silica discs and controls. Nodules formed much earlier on the silica surfaces and these eventually mineralized. Nodule formation was reproducibly enhanced on the silica surface and less markedly on the inverted discs. It is likely that both the surface characteristics of the silica gel and silicic acid release from the disc affect osteoblast behaviour. PMID- 15348932 TI - Dependence of in vitro biocompatibility of ionomeric cements on ion release. AB - The in vitro biocompatibility of a group of ionomeric cements (ICs) was evaluated with respect to their ion release properties. These ICs were made from a defined series of glasses with the general formula 1.5SiO2.0.5P2O5.Al2O3.(1.0 Z)CaO.0.75CaF2 where Z was the mole fraction (ranging from 0-0.1) of an alkali metal oxide, either sodium or potassium or a mixture of both. For these alkali metal ICs, the amount of sodium released was directly related to the sodium content of the constituent glass. Similarly, the amount of potassium released was directly related to the potassium content. There was no correlation between the aluminum content of the glass and the aluminum ion release. Increasing the monovalent cation concentration, however, produced ICs with increased fluoride release. The biocompatibility of the ICs, as assessed by in vitro cell growth and viability measurements, was inversely proportional to aluminum ion release. Fluoride ion release, although important in terms of in vitro biocompatibility, would appear to be less important than aluminum ion release in determining the overall biocompatibility of the ICs studied. PMID- 15348933 TI - Three-dimensional matrices of calcium polyphosphates support bone growth in vitro and in vivo. AB - Novel macroporous calcium polyphosphate (CPP) scaffolds, with three-dimensional interconnected structure, were fabricated using a polyurethane sponge method. They were then employed in both in vitro and in vivo assays to examine their suitability as bone tissue engineering scaffolds. In the former, subcultured rat marrow cells were seeded on the scaffolds at 7.0 x 10(5) cells/sample and cultured for 2 wk. Cell-free controls were employed to monitor changes in the scaffold itself. In the in vivo assay, CPP rods were implanted in rat distal femur and recovered after 2 wk. Samples were examined by scanning electron microscopy following freeze-fracturing. Both in vitro and in vivo assays demonstrated the growth of bone within the scaffolds. In vitro, the bone/CPP interface was occupied by a morphologically distinguishable cement line, while in vivo non-mineralized fibrous tissue was seen at the interface together with bone ingrowth into the scaffold microporosity. The morphology of the individual surface grains of the CPP scaffolds employed in vivo changed to a more rounded form, while no change in geometry was observed in the in vitro cell-free group. These preliminary studies indicate that three-dimensional CPPs can be successfully used as scaffolds for bone tissue engineering. PMID- 15348934 TI - A novel method to examine the phenotype of chondrocytes. AB - Tissue engineering of articular cartilage in order to restore the function of degenerated, diarthrodial joints is currently widely under investigation. The results obtained thus far indicate that proper control of the differentiation of the cells used for this purpose is essential to produce and maintain a hyaline like matrix. In this study, a procedure is described by which differentiation of chondrocytes in vitro and ex vivo can be studied. The method involves quantitative assessment of mRNA for different collagens, which are markers for differentiation of chondrocytes, by competitive PCR. In a culture system employing human osteoarthritic chondrocytes, mRNAs for the alpha1-chains of collagen types I, II and X are quantified. The procedure is fast, specific and sensitive. However, several controls should be included to ascertain the reliability of the assessment. PMID- 15348935 TI - Positively charged polymer polylysine-induced cell adhesion molecule redistribution in K562 cells. AB - We have recently demonstrated that if human K562 erythroleukemic cells, which normally grow in suspension, are grown in polylysine-coated culture flasks for 48 h, they adhere to these flasks and grow in an anchorage-dependent like manner. Important changes in both membrane conductivity (ionic transport across the cell membrane) and membrane permittivity (static distribution of charges across the cell membrane) were also observed, indicating perturbations in membrane lipids, proteins and polysaccharides. In order to better understand the changes occurring in K562 cells exposed to polylysine and because of the important role played by cell adhesion molecules (CAMs) in cell/cell and cell/substratum interactions, and in cellular adaptation to the surrounding environment, the possible redistribution of these molecules after exposure to polylysine were investigated. In particular, the CD54 (ICAM-1), CD58 (LFA-3) and alphaVbeta3 (vitronectin receptor) molecules were investigated at different times of growth both quantitatively and qualitatively utilizing flow cytometry and immunofluorescence microscopy, respectively. The data indicate that there were no significant quantitative variations in the CAMs examined at all the times tested. In addition, no qualitative changes were observed at 48 h (as well as 24 h) of exposure. However, shorter treatment times (30 min, 1 and 2 h) did induce important CAM reorganization. The results seem to demonstrate that this cycle of CAM redistribution may, in part, be responsible for cellular adaptation to the new growth environment of K562 cells and for the variations in membrane electrical properties observed. PMID- 15348936 TI - Comparison of the cytotoxicity of molybdenum as powder and as alloying element in a niobium-molybdenum alloy. AB - Commercially pure metal niobium (c.p. Nb) as well as niobium-molybdenum (Nb-Mo) alloys were produced following several powder metallurgical routes. In brief, niobium and molybdenum powders were blended and milled in order to form Nb-Mo alloys. The alloy powders and the c.p. Nb were then either pressed and sintered, or cold isostatically pressed followed by hot isostatically pressing. In order to assess the cytotoxicity of the c.p. Nb and c.p. Mo powders, a 72 h minimal essential medium-extraction test was performed according to ISO/EN 10993-5. The cytotoxicity of the c.p. Nb metal and the Nb-Mo alloys was tested in a 72 h direct contact test. Compared to a negative control (UHMWPE), c.p. Nb was non toxic, but c.p. Mo was moderately toxic. None of the powder metallurgically produced materials were toxic. Neither differences in molybdenum concentration, nor in porosity of the samples, due to different production routes, had any influence on the toxicity of the materials. Rat bone marrow cultures showed that only on c.p. Nb was a mineralized extracellular matrix formed, while on the more porous Nb-Mo alloys, cell growth was observed, but no mineralization. In conclusion, c.p. Mo powder is moderately toxic, however, as an alloying element it is non-toxic. Material porosity seems to influence differentiation of bone tissue in vitro. PMID- 15348937 TI - Platelet reactions to modified surfaces under dynamic conditions. AB - The influence of surfaces on the reactions of platelets in whole blood under laminar flow was investigated in a cone and plate viscometer. Citrated whole blood was exposed to steel, PMMA and PMMA modified with PEO at low (500 s(-1)) and high (4000 s(-1)) wall shear rates at room temperature for a period of 100 s. Treated blood samples were fixed with paraformaldehyde, stained with a monoclonal antibody for CD41 (platelet GPIIb/IIIa) conjugated with phycoerythrin and analyzed by flow cytometry. The reactions of platelets (microparticle generation and formation of platelet-platelet, platelet-red blood cell and red blood cell microparticle aggregates) to these environments were quantified. Additionally, the size of platelet-platelet aggregates was assessed. The percentage platelet aggregation and numbers of microparticles generated were independent of surface type at any shear rate. The composition of the aggregates formed was influenced by the surface: at low and high shear rates PMMA caused the generation of platelet-platelet aggregates of the greatest size. The numbers of red blood cell platelet and red blood cell-microparticle aggregates also varied depending on the surface. Fewer red blood cell-platelet aggregates were formed at higher shear rates, whereas the reverse was true for red blood cell-microparticle aggregates. It is concluded that these variations may help to explain the differential effects of surfaces to the induction of distant thrombotic events: microparticles may be protected from loss from the blood stream by their association with red blood cells at high shear rates. PMID- 15348938 TI - Soft tissue response to glycerol-suspended controlled-release glass particulate. AB - Vesicoureteral reflux and urinary incontinence have previously been treated by various means including the endoscopic delivery of injectable bulking materials such as silicone micro-implants, PTFE implants, glass particles, fat and bovine collagen. These first three materials do not degrade and collagen requires frequently repeated injections in order to sustain the restored continence provided. Vesicoureteric reflux in children usually resolves independently before the age of five. Correction is required before this, because treatment by prophylactic antibiotics is frequently unsuccessful in preventing breakthrough infection. The ideal material for injection should have large particles to avoid migration, inject easily and controllably, be non-toxic and dissolve over the period of time by which time the kidney will be mature. Three different controlled-release glass (CRG) granule compositions have been prepared by Giltech Ltd, and suspended in a suitable carrier medium (in this case glycerol). The degradable glasses, which have two different size ranges of 200-300 and <53 microm, and three different solution rates, were injected intramuscularly into the dorso-lumbar region of rats. Histological analysis of cryostat cut section after time periods of 2 d, 4 and 9 wk, and 6 mon has been performed. Histology sections were stained for neutrophils and macrophages using enzyme histochemistry. ED1 (monocytes and immature macrophages), ED2 (mature tissue macrophages), CD4 (helper/inducer T-lymphocytes and macrophages), CD8 (suppressor/cytotoxic T-lymphocytes), Interleukin-1beta, IL-2 (activated T lymphocytes), Major Histocompatibility Complex (MHC) class II (activated macrophages and activated B-lymphocytes), alpha-beta (T-lymphocytes) and CD45RA (B lymphocytes) antibodies have been used to stain immunohistochemically each sample. This study demonstrates that particulate, degrading glass is stimulating an inflammatory response in soft tissue at time periods up to 6 mon. It should be noted that very small particulate, fast degrading glass is leading to tissue necrosis and should not be considered further for these applications. However, larger particulate, slower degrading materials are demonstrating effective potential for stress incontinence applications. PMID- 15348939 TI - Synthesis and characterization of carbonate hydroxyapatite. AB - Substituted apatite ceramics are of clinical interest as they offer the potential to improve the bioactive properties of implants. Carbonate hydroxyapatite (CHA) has been synthesized by an aqueous precipitation method and precipitates with two different levels of carbonate, processed as powders. Sintering experiments were performed to establish the influence of carbonate in significantly reducing the temperature required to prepare high-density ceramics when compared with stoichiometric hydroxyapatite (HA). High-temperature X-ray diffraction was used to characterize the phase stability of the apatites on sintering. Increasing carbonate content was shown to reduce the temperature at which decomposition occurred, to phases of CaO and beta-TCP. Mechanical testing, performed using biaxial flexure, showed that the CHA specimens had strengths similar to stoichiometric HA. PMID- 15348940 TI - In vitro real-time characterization of cell attachment and spreading. AB - A method based on the piezoelectric quartz crystal microbalance (QCM) technique for in vitro real-time characterization of cell attachment and spreading on surfaces has been developed. The method simultaneously measures the resonant frequency, f, and the dissipation energy, D, of the oscillating system. The QCM responses are sensitive to very small amounts (a few hundreds) of cells and highly specific to surface chemical properties. The first results from deposition of cells on two polystyrene surfaces of different wettability in serum-containing medium are reported. It has previously been shown that a decrease in f is related to the degree of cell spreading. In our data it appears that the extent or quality of cell attachment is reflected in an increase in D caused by adhering cells. The combined information from f and D measured by this technique might therefore be useful to probe cell-surface interactions for biomaterials. PMID- 15348941 TI - Synthesis of dahllite through a cement setting reaction. AB - Dahllite is a synthetic carbonated hydroxyapatite with a carbonate ion content similar to bone mineral. The first objective of this study was to investigate the stoichiometric conditions under which dahllite formation occurs in a powder mixture of alpha-Ca3(PO4)2, Ca(H2PO4)2 and CaCO3. The second objective was to identify how these conditions apply to commercially available cement, Skeletal Repair System (SRS), and other alpha-Ca3(PO4)2-based cements currently under investigation. The stoichiometric coefficients were found to be a function of both the percentage of carbonate ions incorporated into the hydroxyapatite structure, and the amount of CO2 released during the reaction. As a consequence, a stability field has been obtained where different initial proportions of the reactants in the powder mixture should give the same reaction product if sufficient CO2 is released into the solution. However, increasing amounts of CaCO3 in the initial mixture have been shown to affect the solution pH in such a way that only partial reaction of the reactants takes place. SRS and other alpha Ca3(PO4)2-based cements have been located inside the stability field and a comparison between their reported setting and hardening properties has been performed. PMID- 15348942 TI - Characterization of coatings for open-heart surgery tubing with heparin and lipid. AB - The aim of this study was to compare qualitative and quantitative methods for coating characterization on internal surfaces of polyvinyl chloride (PVC) tubing used in procedures of extracorporeal circulation. The methods of characterization included optical microscopy (OM) after dyeing with toluidine blue, scanning electron microscopy (SEM), atomic force microscopy (AFM) and energy dispersive spectrometry (EDS). OM after sample dyeing was excellent in allowing early detection of any absence or irregularities of coatings among the used methods. SEM was the most effective in observing the structure and thickness either in heparin coatings or in lipid coatings. AFM provided a good evaluation of the surface topography. A conjunction of all methods is recommended for complete characterization. The quantification methods based on colorimetric tests were efficient in determining the concentrations of heparin and lipid on internal surface tubes, the coating stability and the reproducibility of the results. PMID- 15348943 TI - Identification and mapping of the amorphous phase in plasma-sprayed hydroxyapatite coatings using scanning cathodoluminescence microscopy. AB - The presence and distribution of the amorphous phase is a key factor in the performance and bone-bonding behavior of plasma-sprayed hydroxyapatite coatings. Microanalysis of coatings was conducted with microprobe Raman and scanning cathodoluminescence microscopy. It was confirmed that the darker regions in polished cross sections represent the amorphous phase. The more intense cathodoluminescence emission from the amorphous phase during electron-beam irradiation compared with the crystalline phase was used to detect the two structurally different areas within the sample. By selecting the peak of the emission at 450 nm it was possible to raster the surface with the electron beam and produce a map of the amorphous phase in polished sections, a fracture surface and an as-sprayed surface of the plasma-sprayed coating. Cathodoluminescence microscopy, based on the different light emission from the amorphous phase and hydroxyapatite, is a useful tool for identifying and mapping of the amorphous phase constituent in plasma-sprayed coatings. PMID- 15348944 TI - Controlled release of NSAIDs bound to polyacrylic carrier systems. AB - The synthesis, characterization and properties of new acrylic "polymeric drugs" derived from the NSAIDs agents ibuprofen and ketoprofen are described. The swelling behavior in hydrated medium and the controlled release from hydrophilic copolymers of the NSAIDs derivatives and 2-hydroxyethylmethacrylate, HEMA, are discussed, considering the hydrolytical reactivity of the acrylamide or acrylic ester functional groups, the aromatic or aliphatic structure of the spacer side groups, and the hydrophilic character of the copolymer systems. The results obtained demonstrated that the swelling degree of copolymers depends on the average composition of copolymers, decreasing with the increase of the average fraction of the corresponding acrylic derivative of ibuprofen or ketoprofen in the copolymer system. In addition, polymers which support the NSAIDs active component through aromatic amide links, are more sensitive to hydrolytical processes than those of alkyl ester functions. The results obtained demonstrate that these supports could be applied for direct administration, transdermal, systemic or intra-articular injection, as well as in the form of films on wounds. PMID- 15348945 TI - Peripheral nerve regeneration through P(DLLA-epsilon-CL) nerve guides. AB - P(DLLA-epsilon-CL) nerve guides can be used perfectly for short nerve gaps in rats, and are even better than short autologous nerve grafts. The tube dimensions, such as the internal diameter and wall thickness, are very important for the final outcome of peripheral nerve regeneration, as well as the recovery of nerve function. Before using biodegradable nerve guides in patients, it will be necessary to control the swelling the biomaterial during degradation. PMID- 15348946 TI - Interactions of chondrocytes with methacrylate copolymers. AB - Copolymers of poly(ethylmethacrylate) (PEMA) and tetrahydrofurfurylmethacrylate (THFMA) have been shown to exhibit potential as a biomaterial for use in cartilage repair. However, the interactions of chondrocytes with the polymer surface is not well understood. A series of novel methacrylate copolymers containing PEMA, THFMA and hydroxyethylmethacrylate (HEMA) were prepared and the ability of these various copolymers to support chondrocytes attachment in vitro has been assessed by the Alamar blue assay for cell number and environmental scanning electron microscopy (ESEM). As the mole fraction of HEMA in PEMA/THFMA/HEMA copolymers increased, chondrocyte attachment to the polymer surface in 24 h decreased. Chondrocytes maintained a rounded morphology and were strongly attached on the THFMA/PEMA polymer surface, but as the mole fraction of HEMA increased the cells present became much smaller with fewer cell to cell interactions. The effect of pre-adsorbing fibronectin on to the polymer surface on cell attachment was assessed both in the presence and absence of serum. Chondrocyte attachment was significantly reduced in serum-free medium. Pre adsorption of fibronectin on to the copolymer surface substantially increased cell attachment in all cases. In conclusion, chondrocyte attachment and proliferation on these copolymers may be controlled by changes in the polymer surface chemistry and is highly sensitive to the presence of proteins either in the culture media or pre-adsorbed on to the copolymer surface. PMID- 15348947 TI - Development of a new synthetic bone graft. AB - A process for the replication of bovine cancellous bone in synthetic bioceramic materials for use as artificial bone graft substitutes is described. The process detailed here may be easily implemented to allow production of large numbers of blocks of material, even on a laboratory scale. The graft material has a pore morphology and interconnectivity identical with that of the original cancellous bone used as a starting material. Strength of the material is adequate, and at lower porosity levels it meets the FDA requirements for coralline materials for spinal applications. The synthetic graft is also shown to have excellent fluid retention characteristics, making it a potential carrier for morphogenic agents such as solutions of bone morphogenic protein. PMID- 15348948 TI - New starch-based thermoplastic hydrogels for use as bone cements or drug-delivery carriers. AB - The development of new biodegradable hydrogels, based on corn starch/cellulose acetate blends, produced by free-radical polymerization with methyl methacrylate monomer (MMA) and/or an acrylic acid monomer (AA), is reported. The polymerization was initiated by a redox system consisting of a benzoyl peroxide and 4-dimethlyaminobenzyl alcohol at low temperature. These hydrogels may constitute an alternative to the materials currently used as bone cements or drug delivery carriers. Swelling studies were carried out, as a function of pH and temperature, in buffered solutions. The xerogels were further characterized by Fourier transform-infrared spectroscopy. Tensile and compression tests, and dynamic mechanical thermal analysis were used to assess the mechanical performance of the developed materials. The fracture surfaces were observed by scanning electron microscopy. The developed materials are sensitive to the pH, showing a clear reversible transition in a relatively narrow interval of pH, which is just in the range of physiological conditions. These properties make the materials developed in this study very promising for biomedical applications. Fickian-type diffusion is the mechanism predominant in these systems, except for the composition with a higher concentration of AA, that corresponds to the most desirable kinetical behavior for controlled release (case II-transport mechanism). Furthermore, the results obtained in the mechanical tests are in the range of those reported for typical PMMA bone cements, showing that it is possible to develop partially degradable cements with an adequate mechanical behavior. PMID- 15348949 TI - Comparison between the polymerization behavior of a new bone cement and a commercial one: modeling and in vitro analysis. AB - The polymerization behavior of a new bone cement based on poly(ethylmethacrylate), hydroxyapatite powder and n-butylmethacrylate monomer and a commercial cement have been studied. Polymerization kinetics were analyzed by means of differential scanning calorimetry (DSC). DSC data have been used to evaluate a phenomenological model describing the cure kinetics of this new bone cement. The kinetic model coupled with the energy balance was then used to obtain temperature and degree of conversion profiles in the bone-cement-prosthesis system, under non-isothermal conditions, as function of initial temperature and thickness of the cement. Material properties, boundary and initial conditions and the kinetic behavior were the input data for the numerically solved heat-transfer model. The modeling results have been compared with in vitro results. PMID- 15348950 TI - Thin hydroxyapatite coatings via sol-gel synthesis. AB - Production of hydroxyapatite coatings using an alkoxide-based sol-gel route requires control of solution aging time and heating schedule. 31P nuclear magnetic resonance spectroscopy was used to investigate the changes during aging of the sol and thermal gravimetric analysis employed to study the behavior of the xerogels as a function of temperature, while final products were determined using X-ray diffraction. Results from 31P nuclear magnetic resonance spectroscopy and thermal analysis revealed that sols must be aged for at least 24 h to complete the reaction of the two reactants. Deposition of the sol for coating production will then yield monophasic hydroxyapatite. Coatings produced from sols aged for less than 24 h yielded calcium oxide in addition to hydroxyapatite. Prefiring is necessary to remove most of the residual organic materials. Final heating up to 800 degrees C produces crystallization at 550 degrees C and removal of the remaining organic constituents for the formation of a thin hydroxyapatite layer. PMID- 15348951 TI - In vitro response of osteoblasts to hydroxyapatite-reinforced polyethylene composites. AB - A primary human cell culture model was used to investigate a range of hydroxyapaptite (HA)-reinforced high-density polyethylene (HDPE) composites (HAPEX). These materials are being developed as potential bone-substitute materials. When designing and optimizing a second-generation biomaterial, it is important to achieve a balance between mechanical and biological properties without compromising either. Biochemical and histological parameters have been used to compare the biological response of 20% and 40% volume HA in HDPE. Cellular DNA and incorporation of tritiated thymidine was measured to assess cell proliferation. Alkaline phosphatase (ALP) production was used as a marker of osteoblast phenotype expression. In this preliminary study, osteoblasts cultured on the 20% HAPEX showed a greater increase in the rate of proliferation and osteoblast expression as indicated by an increase in ALP activity compared to the 40% HAPEX over the time period studied. Osteoblast-like cells showed a flattened morphology on both composites and in some cases a greater covering was observed on the 20% HAPEX. These results indicate that the composites may not be identical in terms of bioactivity and that further research on surface topography and physico-chemical properties is required to assess fully the biological response of these composites. PMID- 15348952 TI - Direct activation of mast cells by prosthetic biomaterial particles. AB - IL-4 is a mast cell and T cell produced immune cytokine that is important in the regulation of macrophage function. IL-4 has also been implicated in the induction of foreign body giant cell formation. In patients with long-term joint prostheses, a localized granulomatous inflammation develops in periarticular tissues and other organs where phagocytosis of particulate material from various prosthetic components takes place. In this study we used the inflammatory lesions of the bone-implant interface as a model to investigate the possible production, the frequency and the cellular source of IL-4. 40 samples of the interface membrane obtained from 25 patients undergoing revision of clinically failed implants were analyzed by immunohistochemistry. Cryostat sections were labeled with specific monoclonal antibodies to mast cell products: IL-4, tryptase and the receptor c-kit (CD117). The study has identified a significant level of production of IL-4 by mast cells in all the cases analyzed. There was an apparent difference in the number of mast cells in relation to the histological variants of the interface. The increase in the number of mast cells and IL-4 production was more pronounced in cases with heavy macrophage infiltrate than those exhibiting a predominance of giant cells. The findings imply that the recruitment of mast cell and IL-4 expression precede the granulomatous reaction and may have a role in the induction of a number of immunopathological changes related to mast cell activation by biomaterial particles. PMID- 15348953 TI - In vivo bone formation by human bone marrow cells: effect of osteogenic culture supplements and cell densities. AB - Bone marrow is known to contain a population of osteoprogenitor cells that can go through complete differentiation when cultured in a medium containing appropriate bioactive factors. In this study, porous particles of a calcium phosphate material were seeded with adult human bone marrow cells in the second passage. After an additional culture period of 1 wk in the particles, these hybrid constructs were subcutaneouslly implanted in nude mice with a survival period of 4 wk. The cell seeding densities range from 0-200 000 cells per particle and the cell culture system was designed to investigate the single and combined effects of dexamethasone and recombinant human bone morphogenetic protein 2 (rhBMP-2). The hybrid "material/tissue" constructs were processed for histology and the amount of de novo bone formation was quantified, for each culture condition, by histomorphometric techniques. The relative percentage of mineralized bone formation reached a maximal value of 19.77+/-5.06, for samples cultured in the presence of rhBMP-2 and with a seeding density of 200 000 cells/particle, compared to 0.52+/-0.45 for samples in which no cells had been cultured and had been incubated in culture medium supplemented with Dex and rhBMP-2. For the tested conditions and for the low cell numbers used in this study, rhBMP-2 proved to be an essential bioactive factor to obtain in vivo bone formation by our culture system. The results from this study prove the potential of cultured adult human bone marrow cells to initiate and accelerate de novo bone formation after transplantation into an ectopic site. PMID- 15348954 TI - Bone tissue engineering on calcium phosphate-coated titanium plates utilizing cultured rat bone marrow cells: a preliminary study. AB - The use of osteoinductive in vitro tissue-coated implants in orthopaedic and dental surgery (e.g. revision hip arthroplasty), could result in a better fixation of these implants. However, this tissue engineering technology has only proved to be effective in porous materials and not on flat implant surfaces. In this study we have demonstrated that calcium phosphate-coated titanium plates with a layer of cultured osteogenic cells and their extracellular matrix can initiate bone formation in vivo. Both primary and subcultured rat bone marrow cells were grown on to biomimetic calcium phosphate-coated titanium plates. After 7 d of culture, in the presence or absence of dexamethasone, the implants were subcutaneously implanted in nude mice for 4 wk. Control samples, which consisted of calcium phosphate-coated plates without cultured cells and porous calcium phosphate particles with or without cultured cells, were also implanted subcutaneously. At autopsy, no bone formation could be detected on any of the control samples without cells and samples with subcultured cells, which were primary cultured in medium without dexamethasone. In contrast, clear de novo bone formation could be observed on the calcium phosphate-coated plates and in the porous calcium phosphate particles with primary or subcultured cells, which had been continuously cultured in medium with dexamethasone. These results indicate that this hybrid technology offers great potential for the fixation of flat bone replacement implants (e.g. artificial hips) in inferior bone in the future. PMID- 15348955 TI - Maintain forward motion with your staff. PMID- 15348956 TI - Recover the spirit of nursing. PMID- 15348958 TI - Leadership standards: no room for doubt. AB - Share your personal and professional leadership expectations with your staff members. PMID- 15348959 TI - Measuring change: how technology increases nursing's diversity. AB - As the population becomes more diverse, health care must take steps to ensure all cultural and ethnic groups are well-served according to their unique needs. PMID- 15348960 TI - Offer agency nurses resources. PMID- 15348961 TI - Focus on performance, not paperwork. AB - The Joint Commission shares the 2004 updates to the National Patient Safety Goals. PMID- 15348962 TI - Know the nuances of long-term care. AB - Unique documentation and care requirements exist in a long-term care facility. PMID- 15348964 TI - Countering "brain attacks". AB - Proactive nursing markedly improves CVA patients' outcomes. PMID- 15348966 TI - Break through language barriers. AB - Using specific phrases in Spanish to assess acute pain in non-English speaking Hispanic patients helps provide timely pain assessment and management. PMID- 15348967 TI - Medication errors: ending the blame-game. AB - Here, review an analysis of leadership's reaction to medication errors made by nurses, pharmacists, and physicians. PMID- 15348968 TI - Make your best better with a reporting system. AB - To decrease wait times and increase satisfaction for patients, one Chicago facility implemented an ED written report. PMID- 15348973 TI - Trauma, change in strength of religious faith, and mental health service use among veterans treated for PTSD. AB - One of the most pervasive effects of traumatic exposure is the challenge that people experience to their existential beliefs concerning the meaning and purpose of life. Particularly at risk is the strength of their religious faith and the comfort that they derive from it. The purpose of this study is to examine a model of the interrelationships among veterans' traumatic exposure, posttraumatic stress disorder (PTSD), guilt, social functioning, change in religious faith, and continued use of mental health services. Data are drawn from studies of outpatient (N = 554) and inpatient (N = 831) specialized treatment of PTSD in Department of Veterans Affairs programs. Structural equation modeling is used to estimate the parameters of the model and evaluate its goodness of fit to the data. The model achieved acceptable goodness of fit and suggested that veterans' experiences of killing others and failing to prevent death weakened their religious faith, both directly and as mediated by feelings of guilt. Weakened religious faith and guilt each contributed independently to more extensive use of VA mental health services. Severity of PTSD symptoms and social functioning played no significant role in the continued use of mental health services. We conclude that veterans' pursuit of mental health services appears to be driven more by their guilt and the weakening of their religious faith than by the severity of their PTSD symptoms or their deficits in social functioning. The specificity of these effects on service use suggests that a primary motivation of veterans' continuing pursuit of treatment may be their search for a meaning and purpose to their traumatic experiences. This possibility raises the broader issue of whether spirituality should be more central to the treatment of PTSD, either in the form of a greater role for pastoral counseling or of a wider inclusion of spiritual issues in traditional psychotherapy for PTSD. PMID- 15348974 TI - The psychosocial determinants of depression: a lifespan perspective. AB - This cross-sectional study compares depressive risk factors within and between two Western community cohorts. Six hundred eight primary care patients from the rural population of Farranfore, Ireland, and the suburban population of Penrith, Australia, completed a self-report questionnaire designed to be comprehensive in characterizing the psychosocial profile of patients with depressive symptoms. The survey included the Centre for Epidemiological Study Depression scale, Vulnerable Personality Style Questionnaire, Life Events Inventory, and Social Support Survey. Exposure to a range of early adverse childhood events was also assessed. The 3-month prevalence of depressive symptoms among Farranfore and Penrith respondents was 30% and 35%, respectively. The onset of depressive symptoms was positively associated with unfavorable childhood events, poorly perceived social supports, recent stressful life events, a vulnerable personality style, and previous depressive illness. The study identifies the person's social and internal world as a potent source of depressive risk over the lifespan. PMID- 15348975 TI - A review of studies of the Hamilton depression rating scale in healthy controls: implications for the definition of remission in treatment studies of depression. AB - The Hamilton Rating Scale for Depression (HRSD) is the most commonly used symptom severity scale to evaluate the efficacy of antidepressant treatment. On the basis of an expert consensus panel, an HRSD score of < or = 7 was recommended as a cutoff to define remission. Since that recommendation, little empirical work has been conducted to confirm the validity of this threshold. One approach toward determining a cutoff score for defining remission is to establish the range of values for healthy controls. We therefore conducted a literature review of studies of the HRSD in healthy controls to determine the normal range of values. Studies of the HRSD in healthy control groups were identified in two ways. First, a MEDLINE search for the years 1966 to 2002 was conducted using the key words Hamilton, depression, and controls, and articles were reviewed. Second, the 69 studies included in two review articles written by the authors were examined. We identified 27 studies that included data on the HRSD for 1014 healthy controls. Across all studies, the weighted mean (SD) HRSD score, adjusting for sample size, was 3.2 (3.2; 95% CI, 3.0 to 3.4). HRSD scores were similar in geriatric and nongeriatric samples, and in men and women. Because HRSD scores in healthy controls are more likely to follow a skewed than a normal distribution, based on a mean of 3.2 and a SD of 3.2, at least 84% of healthy controls scored 7 or less on the HRSD, and 97.5% scored 10 or less. Thus, these results can be taken as support for the recommended cutoff of 7 on the HRSD to define remission. The results can also be used for normative comparisons in which posttreatment group mean scores are compared with mean scores from normative samples. PMID- 15348976 TI - Depression with versus without manic features in rapid-cycling bipolar disorder. AB - Depression has been identified as a hallmark feature of rapid-cycling bipolar disorder, although less attention has been paid to the presence of manic features accompanying depression in rapid cyclers. To provide greater information about the extent to which depression arises with or without salient manic features in rapid cycling, we conducted a preliminary study of rapid cycling in outpatients seeking treatment at an academic specialty center for bipolar disorder. Forty DSM IV affectively symptomatic bipolar outpatients with past year DSM-IV rapid cycling underwent systematic evaluation of symptoms and illness characteristics. Manic and depressive symptoms, treatments, and clinical features were rated by standardized scales. Major depression was present in most rapid cyclers (85%), but salient manic features were also evident in half of all depressed rapid cyclers. A lifetime history of suicide attempts was significantly more common in rapid cyclers who presented with major depression plus salient manic features than in those who presented with pure depression or pure mania (p = .033). Antidepressants were being prescribed for approximately one third of depressed rapid cycling patients regardless of the presence of concomitant manic features, whereas mood stabilizers tended to be used less often when manic features accompanied depression. Depression in conjunction with manic symptoms, rather than pure depression alone, may be more common among rapid-cycling bipolar patients who seek treatment. Lifetime suicide risk may be greater among rapid cycling patients whose depression occurs in tandem with manic symptoms. Prescribing habits in the community that favor antidepressants over mood stabilizers may promote further mood destabilization in this population. Further studies with larger sample sizes are needed to affirm these provisional findings. PMID- 15348977 TI - Phenomenology of childhood psychosis: findings from a large sample of psychiatrically referred youth. AB - Our objective was to evaluate the scope and clinical correlates of psychotic phenomena in psychiatrically referred children and adolescents. Subjects were 1657 psychiatrically referred youth (mean age = 10.9 years) evaluated from 1991 to 2002. DSM-III-R diagnoses were obtained through maternal report by using the Schedule for Affective Disorders and Schizophrenia for School-Age Children Epidemiologic Version. Childhood-onset psychosis was defined by the presence of delusions or hallucinations. Childhood-onset psychosis was identified in 8% of psychiatrically referred youth. It was associated with a chronic course and high levels of impairment. Comorbidity with disruptive, mood, and anxiety disorders was very severe, with only one of the 132 identified youth with psychosis not having at least one comorbidity. In conclusion, childhood-onset psychosis in referred youth is common and highly morbid. It remains an important topic of research deserving full clinical and scientific attention. PMID- 15348978 TI - Neuropsychological functioning in first-break, never-medicated adolescents with psychosis. AB - The purpose of the current study was to examine neuropsychological functioning in a group of never-medicated first-break adolescents with psychosis. It is the first report of cognition in a sample of adolescents with psychosis in which all patients were drug-naive. Twenty-nine adolescent patients (mean age = 16.07; SD = 2.00; 15 male and 14 female patients) experiencing their first psychotic episode and 17 age-matched and sex-matched normal volunteers (mean age = 16.88; SD = 2.39; 9 male and 8 female subjects) were recruited and assessed with a neuropsychological battery. Measures of attention, memory, language, executive functioning, perceptual motor processing, and motor speed were obtained. Psychiatric symptomatology, estimated verbal IQ, and parental socioeconomic status were also determined. Patients with psychosis were significantly more impaired than normal volunteers; effect sizes were greatest in the areas of executive functioning, attention, and memory, and significantly smaller in areas of language, perceptual motor processing, and motor speed. The pattern was not altered when differences in verbal IQ and parental socioeconomic status were controlled. Sex and age interactions indicated that younger male patients were particularly impaired. The findings demonstrate neuropsychological deficits in adolescents with psychosis and suggest that cognitive deficits are core symptoms in psychotic disorders. PMID- 15348979 TI - Ethnicity and treatment outcome variation in schizophrenia: a longitudinal study of community-based psychosocial rehabilitation interventions. AB - The purpose of this study was to investigate cross-ethnic variations in prospective treatment outcomes over a period of 12 months from community-based psychosocial rehabilitation interventions for people with schizophrenia. Data were pooled from two similar studies of psychosocial rehabilitation for clients with schizophrenia conducted in the same urban settings. The final sample consisted of 226 people diagnosed with a schizophrenia spectrum disorder, consisting of 108 (48%) white subjects, 81 (36%) African American subjects, and 37 (16%) Latino subjects. Main analyses were performed using hierarchical linear modeling. Analyses indicated that both study groups demonstrated statistically significant rehabilitative improvement over a period of 12 months. Although African American subjects had a significantly slower rate of improvement (compared with white subjects) in social functioning, analyses in several other outcome domains generally revealed that all three ethnic groups showed similar change trajectories over time, with no additional ethnic-related differences reaching statistical significance. With the exception of slower social improvement for African Americans, the findings from this study suggest that community-based psychosocial rehabilitation interventions did not result in significantly poorer outcomes for the ethnic minority groups examined. Several important implications are discussed. PMID- 15348980 TI - A meta-analysis of the effects of written emotional disclosure on the health outcomes of clinical populations. AB - A meta-analysis was conducted to examine the effects of the written emotional disclosure paradigm on health outcomes of people with physical or psychiatric disorders. After nine studies were meta-analyzed, it was determined that expressive writing significantly improved health (d = .19; p < .05). However, this positive relationship (r = .10) was not moderated by any systemic variables because of the nonsignificant test of homogeneity (Qw = 5.27; p = .73). Nonetheless, a planned contrast illustrated that expressive writing is more effective on physical (d = .21; p = .01) than on psychological (d = .07; p = .17) health outcomes (Qb > 10.83; p < .001). One explanation for the small effect size (ES) results and the nonsignificant test of homogeneity may be the small and heterogeneous samples used in some of the studies within this research synthesis. Future research with expressive writing should be tested with randomized controlled trials to increase the likelihood of detecting a larger treatment effect. PMID- 15348981 TI - Evidence that health attributions and symptom severity predict insight in schizophrenia. AB - Although a relationship between insight and symptomatology in schizophrenia has been repeatedly demonstrated, the influence of psychological variables such as coping mechanisms and attributional style is less clear. We evaluated health attributions, subjective resources for coping, symptomatology, general cognitive functioning, and insight among 38 consecutive admissions with DSM-III-R schizophrenia from a geographically defined catchment area. Health attributions accounted for a significant amount of insight even after symptom severity was accounted for and together predicted 32% of variation in insight scores. This study emphasizes the multifactorial nature of insight and the importance of psychological variables in addition to symptomatology. PMID- 15348982 TI - Radiation toxicity to the visual system. PMID- 15348983 TI - Radionecrosis of the inferior occipital lobes with altitudinal visual field loss after gamma knife radiosurgery. AB - A patient had bilateral superior altitudinal visual field defects because of radionecrosis of the inferior occipital lobes after gamma knife radiosurgery for a recurrent atypical cerebellar meningioma. Although radionecrosis of the anterior visual pathway has been well-documented, this is the first report of visual field loss associated with occipital lobe radionecrosis. The treatment dose this patient received is within the range of predicted tolerable radiosurgical dosing, although this patient was at increased risk for radionecrosis secondary to previous external beam radiotherapy. By offering an effective, noninvasive treatment, radiosurgery has changed the management of intracranial lesions. Radiosurgery targets a discrete volume of tissue and relatively spares the surrounding normal tissue. Radiation injury, or radionecrosis, is the only significant complication of radiosurgery (). We present a case of bilateral occipital lobe radionecrosis after gamma knife surgery that resulted in bilateral superior altitudinal defects. PMID- 15348984 TI - Lack of radiation optic neuropathy in 72 patients treated for pituitary adenoma. AB - The incidence of radiation optic neuropathy (RON) after external photon beam radiation therapy for nonfunctioning pituitary adenoma (NFA) is not well-studied. Retrospective review of ophthalmological and imaging data in 72 patients with NFA treated between 1985 and 1998 with external beam radiation therapy after surgery Clinical follow-up after radiation therapy had to be at least 18 months. RON was defined as a sudden and profound irreversible visual loss affecting the optic nerve or chiasm A review of previously published cases of RON was then performed. In our cohort, no patient had RON. A total of 11 adequately documented series reports of RON were found in the medical literature on radiation-treated NFAs. The incidence of RON in NFA from these series is 0.53% (95% CI, 0.26%-0.96%). An additional 14 single RON cases have been reported, bringing the total of adequately documented RON cases to 25. RON is a rare complication after external beam radiation therapy for NFA. PMID- 15348985 TI - Ophthalmic artery occlusion secondary to radiation-induced vasculopathy. AB - A 35-year-old man with neurofibromatosis type 1 (NF1) had a left ophthalmic artery occlusion that caused no light perception OS 28 years after having been treated with external beam radiation therapy for a presumed glioma of the right optic nerve and chiasm. Clinical and imaging findings were consistent with radiation-induced cerebral vasculopathy. This ophthalmic complication has never been reported, despite the common occurrence of severe carotid-ophthalmic artery junction stenosis after radiation in NF1 patients. Even though modern radiation techniques limit collateral damage, this modality should be used with discretion in NF1 patients, given the vulnerability of their immature cerebral vasculature to radiation. PMID- 15348986 TI - Bilateral anterior ischemic optic neuropathy after liposuction. AB - A 30-year-old woman had bilateral anterior ischemic optic neuropathy after undergoing large-volume liposuction. Visual function remained stable over a four month follow-up, with decreased visual acuity and marked constriction of the visual fields. To our knowledge, this is the second reported case of ischemic optic neuropathy in this setting. PMID- 15348987 TI - Evidence for a probable causal relationship between tretinoin, acitretin, and etretinate and intracranial hypertension. AB - With the recognition that vitamin A and isotretinoin may cause intracranial hypertension, the authors reviewed 331 case reports of ocular side effects associated with the three other marketed retinoids: tretinoin, acitretin, and etretinate. The reports were drawn from the National Registry of Drug-Induced Ocular Side Effects, the World Health Organization (WHO), the Food and Drug Administration, and medical journals between 1979 and 2003. There were 21 cases of intracranial hypertension associated with these three retinoids, leading to an inference that they are probably causally related to intracranial hypertension by WHO criteria. The lack of positive rechallenge data precludes the inference of a definite causal relationship to intracranial hypertension by WHO criteria. The inference of an independent causal role of these retinoids is further cautioned by the fact that six patients were concurrently using tetracycline or minocycline. Even so, the data suggest that all retinoids may, in rare instances, cause intracranial hypertension. PMID- 15348988 TI - Third cranial nerve palsy from midbrain neurocysticercosis: repeated exacerbation on tapering corticosteroids. AB - Third cranial nerve palsy is rare in neurocysticercosis and is usually caused by supratentorial or sub-arachnoid lesions with accompanying hydrocephalus or meningitis. We report a patient who presented with third cranial nerve palsy caused by neurocysticercosis involving the midbrain. The patient showed repeated exacerbation of symptoms on tapering corticosteroids. The experience with this patient indicates that tapering of corticosteroids should be performed very slowly in such cases. PMID- 15348989 TI - Elevated intracranial pressure associated with idiopathic retinal vasculitis, aneurysms, and neuroretinitis syndrome. AB - The idiopathic retinal vasculitis, aneurysms, and neuroretinitis (IRVAN) syndrome typically occurs in young patients and may produce multiple retinal macroaneurysms, neuroretinitis, and peripheral capillary nonperfusion. Optic disc edema has been described, but elevated intracranial pressure has not been previously documented. We report a case of a 12-year-old girl who presented with bilateral disc swelling and peripapillary hemorrhage. Brain magnetic resonance imaging (MRI) was normal, but lumbar puncture yielded an opening pressure of 360 mm H2O with normal constituents. Fluorescein angiography delineated saccular aneurysms of the retinal arteriolar vasculature, and IRVAN syndrome was diagnosed. MR venography disclosed poor filling of both transverse venous sinuses. Acetazolamide treatment of 14 months did not alter the fundus findings. IRVAN syndrome may present initially with optic nerve swelling and elevated intracranial pressure with subsequent development of the characteristic retinal vascular abnormalities. PMID- 15348990 TI - Inferior oblique paresis, mydriasis, and accommodative palsy as temporary complications of sinus surgery. AB - A 15-year-old boy had temporary hypertropia, supraduction deficit, ipsilateral mydriasis, and accommodative paresis after bilateral endoscopic ethmoidectomy, bilateral partial inferior turbinectomy, septoplasty, and Caldwell-Luc procedures for chronic sinusitis. Postoperative imaging did not disclose any intra-orbital abnormalities. The patient was treated with oral prednisolone 70 mg/day on a tapering schedule. Within two months, the ophthalmic abnormalities had resolved. This is the second report to describe such findings, which are attributed to damage of the inferior division of the third cranial nerve secondary to manipulation of adjacent ethmoid tissues. PMID- 15348991 TI - Effect of age on the pupillomotor field. AB - To differentiate physiologic variation from visual field loss with pupillomotor perimetry, the effect of age on the normal pupillomotor field must be known. Given the absence of reported data, the authors aimed to analyze the effect of age on the pupillomotor field as measured with light stimuli of different properties. Subjects consisted of 23 healthy volunteers aged 20 to 28 years ("younger subjects") and 20 healthy volunteers aged 50 to 67 years ("older subjects"). Within a field of 20 degrees, a sequence of 25 focal light stimuli was performed repeatedly on a monitor. The pupil light reflex (PLR) was recorded to stimuli of different diameter and luminance under mesopic conditions. The mean amplitude of the PLR was calculated for each stimulus location and condition. Increasing stimulus luminance or size caused a larger PLR amplitude and a steeper decline of the PLR amplitude from the center to the periphery of the pupillomotor field. The older subjects had reduced mean PLR amplitude with a less pronounced decrease of PLR amplitude toward the field periphery. For the peripheral locations, the largest PLR amplitude was found in the temporal superior quadrants. There was considerable intra-individual test-retest variation in PLR amplitudes in younger and older subjects. The PLR is markedly reduced in older compared with younger subjects. Older subjects have a relatively less pronounced central peak of sensitivity. There are intra-individual test-retest variations in PLR amplitude and asymmetries in sensitivity within the normal pupillomotor field at any age. These findings must be considered in interpreting the results of pupillomotor perimetry. PMID- 15348992 TI - Optic nerve enhancement in hypotensive ischemic optic neuropathy. AB - A 57-year-old woman who had hypotension and cardiac arrest during coronary artery bypass grafting developed hypotensive ischemic optic neuropathy with no light perception vision OU. Bilateral mid-orbital optic nerve enhancement was found on magnetic resonance imaging (MRI) eight weeks after surgery. Re-examination 16 weeks after surgery showed no light perception vision, dilated un-reactive pupils, and pale optic discs. Bilateral optic nerve enhancement persisted on MRI. Optic nerve enhancement has been reported commonly in radiation-induced ischemic optic neuropathy, occasionally in arteritic ischemic optic neuropathy, and rarely in nonarteritic ischemic optic neuropathy. It has never been reported in hypotensive ischemic optic neuropathy. PMID- 15348993 TI - Superior oblique myokymia caused by vascular compression. AB - A 49-year-old man had left superior oblique myokymia for eight years. Magnetic resonance images with enhanced spoiled gradient recalled acquisition in the steady state (SPGR) and flow imaging using steady acquisition (FIESTA) disclosed a branch of the superior cerebellar artery lying on the root exit zone of the left trochlear nerve. Posterior fossa craniotomy confirmed the imaging findings. A Teflon pad was placed between the compressing artery and the trochlear nerve. The patient's superior oblique myokymia has completely resolved with a one-year follow-up. Only one such case has been previously reported. This is the first report to display the imaging findings. PMID- 15348994 TI - Bilateral orbital metastases from a neuroendocrine tumor. AB - When neuroendocrine tumors metastasize to the orbit, they usually do so as solitary lesions, sometimes involving an extraocular muscle. A 70-year-old woman with a known neuroendocrine tumor had bilateral painful proptosis, orbital soft tissue swelling, and ophthalmoplegia. Imaging showed masses within all the extraocular muscles. Orbital biopsy disclosed metastatic neuroendocrine tumor cells within the connective tissue. PMID- 15348995 TI - Friendly fire: neurogenic visual loss from radiation therapy. AB - The author's experience and review of the medical literature suggest that radiation-induced neurogenic visual loss presents on average 18 months after treatment and usually after cumulative doses of radiation that exceed 50 Gy or single doses to the visual apparatus of greater than 10 Gy. Visual loss may result from lesions of the disc, retrobulbar segment of the optic nerve, optic chiasm, or retrogeniculate pathways. Magnetic resonance imaging, the best means of demonstrating radiation injury to the visual pathway, may show abnormalities before the loss of vision. The second eye may show clinical manifestations of optic neuropathy many months after the diagnosis in the first involved eye. Spontaneous improvement in visual function may rarely occur. Treatment has been disappointing, but if visual dysfunction is detected early, hyperbaric oxygen might be beneficial. The risk of neurogenic visual loss must be factored into the decision to irradiate the brain. PMID- 15348996 TI - Recent advances in radiation oncology. AB - The field of radiation oncology has experienced dramatic progress in recent years. Advances in areas of tumor delineation, treatment planning, delivery, and verification allow modern radiotherapy to deliver high doses with great accuracy, less patient morbidity, and in a highly individualized manner. A good understanding of what can be achieved with modern radiotherapy is important in ensuring an effective multidisciplinary approach to the management of cancer and other benign, yet rapidly proliferating, lesions. PMID- 15348997 TI - Thomas R. Hedges, Jr: witness to the birth of modern neuro-ophthalmology. PMID- 15348998 TI - Opsoclonus-myoclonus syndrome during pregnancy. PMID- 15348999 TI - Projector light visual fields. PMID- 15349000 TI - Thirtieth annual meeting of the North American Neuro-Ophthalmology Society, Orlando, Florida, March 27 to April 1, 2004. PMID- 15349002 TI - Effects of high-dose intravenous buprenorphine in experienced opioid abusers. AB - Sublingual buprenorphine, a long-acting, partial mu-opioid agonist, is as effective as methadone in the treatment of heroin dependence, with a better safety profile due to its antagonist activity. However, the safety of therapeutic doses (8 to 16 mg) that might be diverted for intravenous (i.v.) use has not been demonstrated. To evaluate the safety and possible ceiling effects of buprenorphine administered i.v. to experienced opioid users, buprenorphine was administered to 6 nondependent opioid abusers residing on a research unit; the doses tested, in separate sessions, were 12 mg buprenorphine sublingual, i.v./sublingual placebo, and escalating i.v. buprenorphine (2, 4, 8, 12, and 16 mg). Physiologic and subjective measures were collected for 72 hours post-drug administration. Buprenorphine minimally but significantly increased systolic blood pressure. Changes in heart rate or oxygen saturation among the 7 drug conditions were not statistically significant. The mean maximum decrease in oxygen saturation from baseline was greatest for the 8-mg i.v. dose. Buprenorphine produced positive mood effects, although with substantial variability among participants. Onset and peak effects occurred earlier following i.v. administration: peak i.v. effects occurred between 0.25 and 3 hours; peak sublingual effects occurred at 3 to 7 hours. Duration of effects varied among the outcome measures. The dose-response curves were flat for most parameters, particularly subjective measures. Side effects were mild except in one participant who experienced severe nausea and vomiting after the 12-mg i.v. dose. Buprenorphine appears to have a ceiling for cardiorespiratory and subjective effects and a high safety margin even when taken by the i.v. route. PMID- 15349004 TI - A double-blind, placebo-controlled study of a flexible dose of venlafaxine ER in adult outpatients with generalized social anxiety disorder. AB - Venlafaxine extended release (ER) is a dual serotonin-norepinephrine reuptake inhibitor previously shown to be effective in the treatment of major depressive disorder and generalized anxiety disorder. This placebo-controlled, multicenter, randomized, double-blind trial examined the efficacy and safety of venlafaxine ER in outpatients with generalized social anxiety disorder. Two hundred seventy-two outpatients were randomly assigned to receive either a flexible dose of venlafaxine ER (75 to 225 mg/d) or placebo for 12 weeks. Venlafaxine ER was statistically significantly more effective than placebo as demonstrated by the Liebowitz Social Anxiety Scale total scores at weeks 4 to 12. Scores of both the Clinical Global Impression-Severity and Clinical Global Impression-Improvement scales showed that venlafaxine ER was significantly more effective than placebo at weeks 4 to 12. In addition, more venlafaxine ER-treated patients achieved CGI Improvement scores of 1 or 2 than placebo-treated patients at weeks 4 to 12, demonstrating a greater percentage of responders to venlafaxine ER treatment. Assessment using the fear/anxiety and avoidance subscales of the Liebowitz Social Anxiety Scale and the Social Phobia Inventory Scale also showed venlafaxine ER to be more effective than placebo at weeks 4 to 12 and 6 to 12, respectively. The Sheehan Disability Inventory showed that patients in the venlafaxine ER-treated group had significantly better outcomes in social life at weeks 4 and 12, and in work at week 12. Adverse events were similar to those reported in studies of venlafaxine ER in depression and generalized anxiety disorder. Venlafaxine ER was safe, well tolerated, and efficacious in the short-term treatment of generalized social anxiety disorder. PMID- 15349005 TI - Cost and effectiveness of venlafaxine extended-release and selective serotonin reuptake inhibitors in the acute phase of outpatient treatment for major depressive disorder. AB - OBJECTIVE: The purpose of this retrospective analysis was to estimate the cost and effectiveness of venlafaxine extended-release (VXR) compared with selective serotonin reuptake inhibitors in the outpatient treatment of major depressive disorder. METHODS: Pooled data from 8, 8-week, randomized, double-blind studies comparing treatment of major depressive disorder with venlafaxine/venlafaxine XR (n = 851), selective serotonin reuptake inhibitors (fluoxetine, paroxetine, fluvoxamine; n = 748), or placebo (4 studies; n = 446) were retrospectively analyzed to determine the economic implications of symptom remission from the perspective of a US third party payer and that of an employer. A decision modeling approach was used to determine cost and effectiveness ratios. RESULTS: Patients on VXR were associated with 22.8 depression-free days versus 18.6 depression-free days with the studied selective serotonin reuptake inhibitors, based on the decision model. Productive and quality-adjusted days were also expected to increase for VXR patients (22.06 vs. 19.34 and 4.56 to 9.36 vs. 3.72 to 7.63), as was the percentage of patients achieving full activity (25.9% vs. 19.6%). The expected cost per patient achieving remission of symptoms was US 1303.94 dollars and US 1514.96 dollars, and the cost per depression-free days was US 25.66 dollars and US 28.25 dollars, for the VXR and selective serotonin reuptake inhibitors groups, respectively. CONCLUSIONS: Treatment with VXR is not only expected to increase the rate of remission of symptoms but is also associated with achievement of full activity, higher number of depression-free days, productive days, and quality-adjusted days. VXR is a cost-effective treatment option for major depressive disorder. PMID- 15349006 TI - Psychosocial functioning during the treatment of major depressive disorder with fluoxetine. AB - BACKGROUND: Major depressive disorder (MDD) is associated with significant disability, having a profound impact on psychosocial functioning. Therefore, studying the impact of treatment on psychosocial functioning in MDD could help further improve the standard of care. METHODS: Two hundred twenty-two MDD outpatients were treated openly with 20 mg fluoxetine for 8 weeks. The self report version of the Social Adjustment Scale was administered at baseline and during the final visit. We then tested for the relationships between (1) self report version of the Social Adjustment Scale scores at baseline and clinical response, (2) nonresponse, response and remission status and overall psychosocial adjustment at end point, (3) the number/severity of residual depressive symptoms and overall psychosocial adjustment at end point in responders, and (4) the time to onset of response and overall psychosocial adjustment at end point. RESULTS: An earlier onset of clinical response predicted better overall psychosocial functioning at end point (P = 0.0440). Responders (n = 128) demonstrated better overall psychosocial adjustment at end point than nonresponders (P = 0.0003), while remitters (n = 64) demonstrated better overall psychosocial adjustment at end point than nonremitted responders (P = 0.0031). In fact, a greater number/severity of residual symptoms predicted poorer overall psychosocial adjustment at end point in responders (P = 0.0011). Psychosocial functioning at baseline did not predict response. CONCLUSIONS: While MDD patients appear equally likely to respond to treatment with fluoxetine, regardless of their level of functioning immediately before treatment, the above results stress the importance of achieving early symptom improvement then followed by full remission of depressive symptoms with respect to restoring psychosocial functioning in MDD. PMID- 15349007 TI - Pharmacotherapy for bipolar disorder and comorbid conditions: baseline data from STEP-BD. AB - Relatively absent from previous studies of the pharmacotherapy for bipolar disorder is examination of the impact of comorbidity on treatment choices. This has occurred despite the presence of high levels of comorbid anxiety and substance use disorders, and the association of these disorders with severity and course markers of bipolar disorder. In this study, we examined comorbid disorders, identified by structured interviews, and the pharmacotherapy reported at study entry by the first 1000 patients entered into a large, multicenter study of bipolar disorder (Systematic Treatment Enhancement Program for Bipolar Disorder). Our study focused on the degree to which comorbid conditions are linked to the reported use of mood stabilizers deemed "minimally adequate" and the association between specific comorbidities and pharmacotherapy treatment, such as the use of anxiolytics in patients with anxiety disorders. Despite the presence of high levels of comorbidity, the presence of these disorders was only minimally associated with pharmacotherapy. Of the sample of bipolar outpatients, only 59% reported pharmacotherapy use meeting criteria for "minimally adequate" mood stabilizer, regardless of comorbid diagnoses, rapid cycling, or bipolar I or II status. Moreover, the cross-sectional use of "comorbidity-specific" pharmacotherapy for anxiety disorders, substance use disorders, and attention deficit disorder in this outpatient sample of patients with bipolar disorders was limited, suggesting that comorbid conditions in patients with bipolar disorder may be undertreated. Our findings highlight the need for greater clinical guidance and treatment options for patients with bipolar disorder and comorbidity. PMID- 15349008 TI - Valproic acid-induced hyperammonemia: a case report. AB - The authors present a case of a patient treated with valproic acid for seizure disorder who presented with acute mental status changes consistent with encephalopathy. Notably, her serum ammonia level was 3 times the upper limit of normal, despite an only mildly elevated aspartate aminotransferase and normal bilirubin. Her serum valproic acid level was in the therapeutic range. Her symptoms resolved with discontinuation of valproic acid and supportive care. The authors review the possible mechanisms of valproic acid-associated hyperammonemia with encephalopathy and propose clinical practice modifications to minimize the incidence of this adverse reaction to this generally well-tolerated and clinically important psychotropic medication. PMID- 15349009 TI - Lipid metabolism and insulin resistance in depressed patients: significance of weight, hypercortisolism, and antidepressant treatment. AB - Major depression increases cardiovascular risk despite lower cholesterol levels. Little is known about effects of antidepressants on metabolic risk factors. We studied lipoprotein composition, insulin sensitivity (quantitative insulin sensitivity check index), and saliva cortisol in 78 depressed patients before and after 35 days of amitriptyline or paroxetin treatment. Data were analyzed by principal component factor analyses and analysis of variance (ANOVA). At baseline, quantitative insulin sensitivity check index was inversely correlated with cortisol (r = -0.46; P = 0.005) in normal weight patients, with body mass index in overweight patients (r = -0.50; P < 0.001). In overweight patients, hypercortisolemia correlated inversely with total and low density lipoprotein cholesterol (eg, cortisol at 4:00 PM and low density lipoprotein cholesterol: r = -0.49, P = 0.002). After treatment, quantitative insulin sensitivity check index was unchanged. Triglycerides increased in responders to amitriptyline only (P < 0.05). Parameters of cholesterol metabolism improved slightly without differences between treatment groups (eg, high density lipoprotein: pre 43.5 +/- 12.0; post 47.6 +/- 13.0 mg/dL; P = 0.01; low density lipoprotein triglycerides, a measure of low density lipoprotein atherogenicity: pre 458 +/- 120; post 415 +/- 130 mg/g; P < 0,01). The inverse correlation of cortisol and cholesterol, at least in the obese subgroup, proposes a mechanism for the known association of depression with low cholesterol. As determinants of plasma lipids in major depression, we identified body mass index, insulin sensitivity, and cortisol. Although uncontrolled, our data suggest that treatment of depression exerts a mainly beneficial effect on lipid regulation. PMID- 15349010 TI - Quetiapine for treatment of alcohol dependence. AB - Quetiapine is an atypical antipsychotic that has sedative effects. In this retrospective study, indices of alcohol use were compared for alcohol-dependent subjects who either were (n = 30) or were not (n = 20) treated with quetiapine (25 to 200 mg nightly) for disturbed sleep. Indices examined included total days of abstinence, number of hospitalizations for detoxification, and days to first relapse over 1 year of clinic treatment. Subjects were male veterans. All subjects had a diagnosis of alcohol dependence, and 90% of subjects in each group were also diagnosed with posttraumatic stress disorder. Both treatment groups contained a large number of subjects treated with psychiatric medications other than quetiapine. Significant differences were not found between the groups with respect to mean age, detoxifications undergone during the previous year, frequency of comorbid posttraumatic stress disorder or depression, or antidepressant use. The mean number of days abstinent was significantly greater, and the number of hospitalizations was significantly lower for the quetiapine than for the control group during the period studied. The mean number of days to relapse approached significance for the quetiapine as compared to the control group. This study has the usual limitations of a retrospective review, including the lack of standardized assessments of alcohol use. The results of this study are consistent with the hypothesis that the use of quetiapine to improve disturbed sleep may help alcohol-dependent patients maintain abstinence, although decreased drinking may also be a result of improving posttraumatic stress disorder symptoms or of a direct action of quetiapine to reduce alcohol use. PMID- 15349011 TI - Early effects of paroxetine on serotonin storage, plasma levels, and urinary excretion: a randomized, double-blind, placebo-controlled trial. AB - Selective serotonin reuptake inhibitors efficiently decrease intraplatelet concentrations of the platelet activator and potent vasoconstrictor serotonin within 2 weeks of treatment. As elevated plasma serotonin levels potentially lead to vascular adverse events, like vasoconstriction, it is of interest to examine whether selective serotonin reuptake inhibitors may acutely increase plasma serotonin levels. Twenty healthy male smoking volunteers received the selective serotonin reuptake inhibitors paroxetine 20 mg/d for 18 days in a double-blind, placebo-controlled, block-randomized, 2-way crossover study to characterize the acute effect of paroxetine on serotonin plasma levels and urinary excretion. Paroxetine decreased intraplatelet serotonin concentrations by a median of 16% after 24 hours and by -93% after 18 days (P < 0.001). After 24 hours, there was a slight transient rise in plasma serotonin concentration by 36%-which ranged within physiologic concentrations of the control period. Concomitantly, urinary serotonin excretion increased by 89% after 24 hours. In conclusion, initiation of paroxetine treatment does not increase plasma concentrations of the potent vasoconstrictor serotonin to a pathologically relevant extent. PMID- 15349012 TI - Efficacy and tolerability of premenstrual use of venlafaxine (flexible dose) in the treatment of premenstrual dysphoric disorder. AB - The objective of this study was to examine the efficacy and tolerability of intermittent dosing of venlafaxine for the treatment of premenstrual dysphoric disorder. One hundred and twenty-four women aged 18 to 45 years, with regular menstrual cycles and who reported significant premenstrual symptoms, were assessed prospectively to confirm their diagnosis of premenstrual dysphoric disorder. Twenty subjects with confirmed premenstrual dysphoric disorder entered a single-blind, placebo phase (1 cycle). Placebo nonresponders (n = 12) received 2 cycles of intermittent (premenstrual) treatment with venlafaxine (75 to 112.5 mg/d). Subjects initiated treatment 14 days before the anticipated onset of menses and discontinued it on the second day of bleeding. Doses could be adjusted after cycle 1 based on subjects' response and tolerability. Response to treatment was assessed based on changes in the Daily Rating Severity of Problems and Premenstrual Tension Syndrome Questionnaire scores from baseline (before the placebo cycle), as well as Clinical Global Impression-Severity scores. Discontinuation symptoms were assessed between treatment cycles, using the Discontinuation-Emergent Signs and Symptoms questionnaire. Eleven subjects concluded 2 cycles of intermittent dosing with venlafaxine. Nine subjects (81.8%) showed satisfactory response based on Clinical Global Impression of < or = 2. Changes in Daily Rating Severity of Problems scores and subscores (depression, physical symptoms, and anger) and in Premenstrual Tension Syndrome Questionnaire scores were significant (P < 0.05 for all comparisons, Wilcoxon tests). Intermittent treatment was well tolerated. This preliminary report suggests that premenstrual use of venlafaxine is an efficacious and well-tolerated treatment for premenstrual dysphoric disorder. PMID- 15349013 TI - Evaluating the economic consequences of early antidepressant treatment discontinuation: a comparison between controlled-release and immediate-release paroxetine. AB - INTRODUCTION: Early antidepressant discontinuation has been linked to significant clinical and economic consequences. Clinical practice guidelines suggest that treatment should last for at least 3 to 9 months into the continuation phase; however, 30% of patients discontinue therapy within 30 days, and over 40% discontinue therapy within 90 days of initiation, primarily due to adverse events. Clinical trials have shown that controlled-release (CR) paroxetine has a favorable tolerability profile when compared to immediate-release (IR) paroxetine, which may result in lower discontinuation rates and improved economic outcomes. This is the first study to directly compare treatment discontinuation rates and health care expenditures of a CR selective serotonin reuptake inhibitors with its IR counterpart. METHODS: This matched retrospective study used claims from a national managed care database to assess differences in discontinuation rates and health care expenditures between paroxetine CR and IR for treating depression and/or anxiety. Discontinuation was assessed by survival analysis, and health care expenditure was assessed using average monthly medical and pharmacy charges. RESULTS: There were 1275 paroxetine CR patients and 2550 paroxetine IR patients matched in the analysis. At 90 days, 62% of paroxetine CR patients continued therapy versus 56% of paroxetine IR patients. At 180 days, 51% of paroxetine CR patients continued therapy versus 42% of paroxetine IR patients. When evaluating all medical charges, paroxetine CR patients incurred US 119 dollars less per month than paroxetine IR patients (P = 0.054). CONCLUSIONS: Patients receiving paroxetine CR remained on therapy longer than patients on paroxetine IR, which resulted in lower total monthly medical costs for patients receiving paroxetine CR. Differences in costs were primarily driven by reduction in hospitalization expenditures. PMID- 15349014 TI - Fluvoxamine and graded psychotherapy in the treatment of bulimia nervosa: a randomized, double-blind, placebo-controlled, multicenter study of short-term and long-term pharmacotherapy combined with a stepped care approach to psychotherapy. PMID- 15349015 TI - Reply to "Risperidone-induced obsessive-compulsive symptoms: serotonin-dopamine imbalance?". PMID- 15349016 TI - A prospective multicentric trial for effectiveness and tolerance of a N2O/O2 premix used as a sedative drug. PMID- 15349017 TI - Effectiveness of orlistat versus diet and exercise for weight gain associated with antidepressant use: a pilot study. PMID- 15349018 TI - Adjunctive risperidone treatment in combat veterans with chronic PTSD. PMID- 15349019 TI - The changing pattern of inpatient antipsychotic drug use in Italy. PMID- 15349020 TI - The reliability of the Mini-International Neuropsychiatric Interview--Italian version. PMID- 15349021 TI - Acute dystonic reaction with moderate-dose ziprasidone. PMID- 15349022 TI - SSRI-induced enuresis: a case report. PMID- 15349023 TI - Topiramate can cause lithium toxicity. PMID- 15349024 TI - Mania induction associated with atomoxetine. PMID- 15349025 TI - Elevated quetiapine serum concentrations in a patient treated concomitantly with doxepin, lorazepam, and pantoprazole. PMID- 15349026 TI - Neurologic soft signs and olanzapine treatment in chronic schizophrenia. PMID- 15349027 TI - Olanzapine-induced clitoral priapism. PMID- 15349028 TI - Chronic interictal psychosis responsive to aripiprazole. PMID- 15349029 TI - Baclofen as adjunctive treatment for a patient with cocaine dependence and schizoaffective disorder. PMID- 15349030 TI - The role of environmental tobacco smoke in genetic susceptibility to asthma. AB - PURPOSE OF REVIEW: The increase in asthma prevalence over the past 50 years suggests that exposures to environmental risk factors have also increased during this time. Environmental tobacco smoke is one of the most common indoor air pollutants and has been associated in epidemiologic studies with airway and allergic phenotypes in exposed individuals. However, symptoms occur in only some individuals, suggesting that individual genotypes determine sensitivity to environmental tobacco smoke exposure. In this review, we summarize studies evaluating the relationship between genotype, environmental tobacco smoke exposure and risk for asthma and related phenotypes. RECENT FINDINGS: Using either candidate gene or genome-wide approaches, a number of studies have examined interactions between genotypes at specific loci or genome regions and environmental tobacco smoke exposure and risk for asthma or asthma-associated phenotypes. These studies implicate variation in the genes encoding the alpha(2) adrenergic receptor, interleukin-10, glutathione S-transferase M1, interleukin-1 beta and interleukin-1 receptor antagonist, matrix metalloproteinases 1 and 12, interleukin-4 receptor alpha-chain, alpha(1)-antitrypsin, and microsomal epoxide hydrolase, as well as unknown genes on chromosomes 1p, 5q and 17p as contributing toward susceptibility in smoking exposed individuals. SUMMARY: Considering environmental tobacco smoke exposure in genetic studies may help to identify more homogeneous subsets of patients that share a common disease etiology. By stratifying samples by environmental tobacco smoke exposure, associations or linkages with specific polymorphisms or chromosomal region may be revealed, as illustrated in the studies discussed in this review. PMID- 15349031 TI - Traffic related pollutants in Europe and their effect on allergic disease. AB - PURPOSE OF REVIEW: The prevalence and incidence of allergic diseases have increased in Europe during the last decades, as in most industrialized countries in other parts of the world. Persistent exposure to traffic related air pollution and especially particulate matter from motor vehicles has often been discussed as one of the factors responsible for this increase. This view seems to be supported by recent human and animal laboratory-based studies, which have shown that particulate pollutants, and in particular diesel exhaust particles, can enhance allergic inflammation and induce the development of allergic immune responses. However, the results from epidemiologic research provide a more complex picture. RECENT FINDINGS: It has been clearly shown in many studies that traffic related air pollution contributes to increased mortality risk; in particular in relation to cardiopulmonary causes. Traffic related air pollution also increases the risk of non-allergic respiratory symptoms and disease. However, for allergic symptoms and illnesses like asthma, allergic rhinitis, atopic dermatitis, wheeze, and allergic sensitization less consistent results have been found. This is the reason why the World Health Organisation concludes cautiously that traffic related air pollution may increase the risk of allergy development and may exacerbate symptoms in particular in susceptible subgroups. This review concentrates on recent epidemiologic studies on the long-term effects of exposure to traffic related air pollution on allergic disease in Europe. SUMMARY: In conclusion, the evidence for an increased risk for asthma and hay fever still is weak but seems to be strengthened a little. However, many questions are left open. PMID- 15349032 TI - Genetic susceptibility to ozone-induced lung inflammation in animal models of asthma. AB - PURPOSE OF REVIEW: Epidemiological associations between ozone exposure and allergic responsiveness are well-documented and have been corroborated in animal studies. The complex interaction between ozone and allergen has genetic and environmental components that affect atopic individuals and may increase the incidence of allergy in susceptible individuals. This review describes the advances that have been made in understanding mechanisms of genetic susceptibility to ozone-induced inflammation, and the interaction between ozone and allergen exposure in mice and a non-human primate model. RECENT FINDINGS: Antioxidant and innate immune defense genes contribute to ozone-induced inflammation and hyperpermeability in mice and humans. Ozone exposure during the allergic challenge phase induces greater enhancement of allergic responsiveness than the sensitization stage. Ovalbumin-pulsed dendritic cells injected into naive mice successfully sensitize the mouse to ovalbumin in the absence of adjuvant. Debate continues over the role of T helper 1-T helper 2 immune profile development in mediating the ozone-allergen interaction, and the potential confounding influence of the predominant T helper 2 system most commonly used to study these responses. SUMMARY: The role of genetic background in susceptibility to ozone-induced lung inflammation has been confirmed, and promising candidate genes have been identified. Descriptive studies confirm that ozone exacerbates allergic responsiveness. Ozone administered during the challenge phase of ovalbumin allergen exposure induces greater responsiveness than during the sensitization phase. Allergen-induced responses enhanced by concurrent ozone exposure warrant further mechanistic research, particularly regarding the influence of susceptibility genes. PMID- 15349033 TI - Diesel exhaust particles and their effect on induced cytokine expression in human bronchial epithelial cells. AB - PURPOSE OF REVIEW: It has been demonstrated that particulate air pollutants such as diesel exhaust particles (DEPs) are associated with allergic respiratory disorders, including asthma and allergic rhinitis. Recent in-vivo and in-vitro studies strongly suggest that DEPs induce both anti-inflammatory and proinflammatory products by activating their transcription. If pollutants are to be controlled in the environment in a cost-effective manner, it is important that the molecular target(s) of DEP-induced responses be elucidated. Bronchial epithelial cells are the key regulators of airway inflammation, and therefore it is crucial to clarify the cellular and molecular mechanisms that are active in these cells. It is also important to compare responses to DEP exposure between healthy and susceptible populations. RECENT FINDINGS: Studies have shown that DEPs activate several signalling pathways such as mitogen-activated protein kinases and transcription factors, in particular nuclear factor-kappa and activator protein-1, in human bronchial epithelial cells; these effects are thought to occur via production of reactive oxygen species. Antioxidants effectively reduce the allergic inflammatory effects of DEPs both in vitro and in mice. SUMMARY: Host responses to DEPs are regulated by a balance between antioxidants and proinflammatory responses. DEP-induced oxidants play a key role in proinflammatory reactions. Recent human studies suggest that chemoprevention against DEP-induced adverse effects in susceptible individuals is possible. PMID- 15349034 TI - Bioaerosols and innate immune responses in airway diseases. AB - PURPOSE OF REVIEW: We review the role of bioaerosols in the pathogenesis of inflammatory airway disease. The focus is on recent discoveries in innate immune responses induced by common components of bioaerosols. RECENT FINDINGS: Common components of bioaerosols include endotoxin, peptidoglycan and beta-glucan; all of which have been associated with inflammatory airway disease. Endotoxin signaling through toll-like receptor 4 is well characterized and updated. Peptidoglycan is now known to signal through three types of molecules: toll-like receptor 2; peptidoglycan recognition proteins; and nucleotide-binding oligomerization domain molecules. Beta-glucan, a common fungal cell wall component, signals through the newly discovered receptor, dectin-1. Emerging data indicate that genetic polymorphisms influence the response to bioaerosols. SUMMARY: Activation of the innate immune system by bioaerosols is becoming better understood. This knowledge provides an opportunity to better prevent and treat airway diseases that result from environmental exposure. PMID- 15349035 TI - Autoimmunity and atopic dermatitis. AB - PURPOSE OF REVIEW: It has been demonstrated that a considerable percentage of patients suffering from atopic dermatitis mount IgE autoantibodies against a broad variety of human proteins. This review summarizes evidence for autoimmune mechanisms in atopic dermatitis and suggests novel pathomechanisms that may be involved in this disease. RECENT FINDINGS: It has been shown that patients suffering from atopic dermatitis exhibit IgE autoreactivity to human proteins. These autoantigens are expressed in a variety of cell and tissue types. Complementary DNAs coding for IgE autoantigens have been identified, cloned and characterized at the molecular level. Using purified recombinant IgE autoantigens, it has been shown in paradigmatic models that IgE autoimmunity may be a pathogenetic mechanism in atopic dermatitis. Moreover, it has been shown that the levels of IgE autoantibodies are associated with severity of disease. SUMMARY: Patients suffering from severe manifestations of atopy mount IgE autoantibodies against a variety of human proteins. The levels of IgE autoantibodies correspond with disease severity. Several mechanisms of IgE autoimmunity may contribute to the pathogenesis of atopic dermatitis. PMID- 15349036 TI - Microbial colonization and atopic dermatitis. AB - PURPOSE OF REVIEW: Atopic dermatitis is a chronic relapsing, pruritic inflammation of the skin, affecting 10-20% of children and 1-3% adults worldwide, with increasing prevalence in highly industrialized countries. Here we review relevant studies, published since June 2002, about immunological triggers in atopic dermatitis, with emphasis on the role of microbial colonization. RECENT FINDINGS: During the past 2 years there has been considerable interest in the mechanisms and trigger factors underlying the increased microbial colonization of atopic skin. Staphylococcus aureus appears to play a significant role as it leads to a worsening of disease severity by producing superantigens that induce a strong proliferation of T cells and favour a T helper type 2-like cytokine profile. In addition, different Malassezia species seem to elicit and maintain skin inflammation after sensitization, but the precise immunological pathway has not yet been described. All these microorganisms are not only perceived as aetiological factors but also as agents responsible either for sustained disease activity or resistance to therapy by modulation of the immune response. SUMMARY: New insights into the important role of microorganisms and their key immunomodulatory pathways in atopic dermatitis may have important implications from a therapeutic point of view because patients with atopic dermatitis may benefit from more than just anti-inflammatory treatment in the future. PMID- 15349037 TI - Role of food allergy in atopic dermatitis. AB - PURPOSE OF REVIEW: Food allergy and atopic dermatitis often occur in the same patient. Based on clinical data from the past few decades, it is clear that foods such as cow's milk and hen's eggs can directly provoke flares of atopic dermatitis particularly in sensitized infants, whereas inhaled allergens and pollen-related foods are of greater importance in older children, adolescents and adults. Because the role and immunology of food allergy in atopic dermatitis remain controversial, here we review data that mainly focus on skin eczema and food allergy. RECENT FINDINGS: Clinical studies have revealed that more than 50% of all children with atopic dermatitis that can be exacerbated by certain foods will react with a worsening of skin eczema alone or in addition to immediate symptoms. Adolescents and adults also react to foods, but reactions to 'classical' food allergens such as hen's eggs and cow's milk are not as common as in childhood. Subgroups of children and of adults with atopic dermatitis do, however, react to pollen-associated foods. Both immunoglobulin E associated and independent T-cell mediated responses appear to be involved in clinical eczematous reactions. SUMMARY: Food-induced eczema should be diagnosed only by a thorough diagnostic procedure, taking into account the patient's history, the degree of sensitization and the clinical relevance of the sensitization. More clinical and immunological studies are needed to unravel the pathophysiology and the different rates of clinical responsiveness to different foodstuffs in patients with atopic dermatitis. PMID- 15349038 TI - Chronic urticaria and infections. AB - PURPOSE OF REVIEW: The pathogenesis of chronic urticaria is multifactorial and a specific treatment is lacking. In acute urticaria there is no doubt of a causal relationship with infections and all chronic urticaria must start as the acute form. However, in the chronic form a primary role for infection is controversial, although it is undeniable that concurrent infections exacerbate the condition. This is the first English language review based on a detailed analysis of current peer-reviewed publications dealing with infections and chronic urticaria. RECENT FINDINGS: In chronic urticaria there is a lot of evidence for different infections, but randomized controlled trials are missing. The prevalence of infections is not increased but in susceptible patients the immune response may lead to the development of chronic urticaria. Interestingly, there is evidence for an infection-associated autoreactive response at least in the subgroup with a positive autologous serum skin test. A variety of mechanisms have been invoked to explain these observations, including molecular mimicry. SUMMARY: Actually the arguments for an important role of underlying causal infections in chronic urticaria are weak, from an evidence-based viewpoint, but there are data suggesting a link. Moreover, an association with underlying or precipitating infectious causes is difficult to establish because there is no possibility for challenge and the number of other urticarial triggers is vast. For the future it will be necessary to reveal the link between urticaria, autoreactivity, non immunoglobulin E-mediated hypersensitivity reactions and infections to find attractive and specific therapeutic interventions for urticarial symptomatology. PMID- 15349039 TI - Latex allergy and skin. AB - PURPOSE OF REVIEW: In this review we address the prevalence and outcome of latex allergy in health care workers (HCWs). Recent findings in natural rubber latex (NRL) allergens and trials of specific immunotherapy (SIT) are also of interest. RECENT FINDINGS: A study involving skin prick test (SPT) screening in HCWs in Russia and adjacent countries found a prevalence of latex allergy of 1.9%. Questionnaire studies performed in Wales and in the USA identified prevalence rates of about 0.6%. An intervention undertaken at the Mayo Clinic, in which only gloves with low or undetectable allergen levels were allowed, reduced markedly the incidence of NRL allergy. Two studies, one from Finland and another from Ohio, showed that outcomes in latex-allergic HCWs are generally good. A study involving SPT screening showed that 6% of construction workers had latex allergy. A questionnaire study among allergists practicing in the USA showed that 62% performed latex SPT and 6% reported anaphylaxis, which mostly occurred while using a homemade SPT solution. Hev b 2, Hev b 5, Hev b 6.01 and Hev b 13 produced positive SPT reactions in over 60% of latex-allergic individuals. Topical application of NRL in a murine model of protein contact dermatitis caused a striking increase in prohevein-specific (Hev b 6.01) immunoglobulin E levels, together with a T-helper-2 type dermatitis. A placebo-controlled SIT trial with NRL extract alleviated cutaneous symptoms but caused some systemic reactions in latex-allergic patients. SUMMARY: Low prevalence rates suggest that the peak of the latex allergy epidemic has already passed in HCWs. Hospital-wide interventions requiring use of low-allergen gloves reduce sensitization and changing gloves to nonlatex ones, or even using low-allergen latex gloves, in the affected individuals appears to confer adequate secondary prevention. In the USA there is an urgent need for standardized latex SPT reagent. Hev b 5 and Hev b 6.01 are major in vivo NRL allergens. Findings in mice suggest that NRL proteins eluting from latex gloves may also cause hand eczema in humans. SIT with NRL extract must still be considered an experimental treatment. PMID- 15349040 TI - Occupational allergens in dentistry. AB - PURPOSE OF REVIEW: Dental professionals and their physicians frequently do not recognize, accurately diagnose or appropriately manage occupational allergies. Dental allergen identification, diagnostics and practical avoidance strategies are summarized in this review. RECENT FINDINGS: Methacrylates, natural rubber latex proteins, rubber glove allergens, and glutaraldehyde are the predominant allergens in dentistry. Reactions range from cell-mediated contact allergy to urticaria and occupational asthma. SUMMARY: Despite recent advances in allergen characterization and increased awareness of selected allergens, treatment of occupational allergies can be improved. Better information and improved cooperation between dental workers and their clinicians is needed. PMID- 15349041 TI - The immunological and clinical spectrum of delayed drug-induced exanthems. AB - PURPOSE OF REVIEW: Drug-induced exanthems are the most common manifestations of drug hypersensitivity and are observed in as much as 2-3% of hospitalized patients. Here we summarize new concepts of the immune mechanisms underlying various forms of drug-induced exanthems. RECENT FINDINGS: Alpha-betaTCR+, CD4 and CD8+ T cells are involved in different drug hypersensitivity reactions. Their function determines the clinical picture. In maculopapular, bullous and pustular exanthems cytotoxic T cells are involved, while a high IL-5 and eotaxin production by tissue cells is frequently found in maculopapular and occasionally in bullous and in pustular exanthems. High IL-8 (CXCL-8) and granulocyte macrophage colony stimulating factor production by T cells is a hallmark of pustular drug exanthems. In the most severe and potentially life-threatening forms of exanthems (Stevens-Johnson syndrome/toxic epidermal necrolysis) cytotoxic CD8+ T cells with natural killer cell markers can be found in the blister fluid. SUMMARY: These findings are the basis for a new subclassification of delayed, type IV hypersensitivity reactions into type IVa (T helper type 1 cells, e.g. tuberculin reaction and contact dermatitis), IVb (T helper type 2 cells, maculopapular exanthem with eosinophilia), IVc (cytotoxic T cells, contact dermatitis, maculopapular and bullous exanthem), and IVd reactions (CXCL 8/granulocyte-macrophage colony stimulating factor-producing T cells and neutrophil attraction, pustular exanthems), by which, in most reactions, various mechanisms occur together but one reaction dominates the clinical picture. PMID- 15349042 TI - Ocular allergy and dry eye syndrome. AB - PURPOSE OF REVIEW: Ocular allergy is a common clinical disorder that includes dry eye syndrome in its differential diagnosis. While ocular allergy treatments have continued to evolve since the early 1990s when the new prescription topical agents became available, there have been no major advances in the treatment of dry eye syndrome other than changes in the chemical structures of various artificial tear formulations. This review is timely and relevant due to the recent FDA approval of several new agents for the treatment of dry eye syndrome. RECENT FINDINGS: The literature reviewed brings the practicing allergist/clinical immunologist up to date on the recent understanding that T-cell activation plays a key role in dry eye syndrome immunopathophysiology. In addition, the parallel novel treatment developments are discussed, including new formulations for tear substitutes, topical cyclosporine A and purinergic receptor (P2Y2) agonists. SUMMARY: The recent developments bode well for patients who are referred for ocular allergy, including dry eye syndrome. A new formulation for a tear substitute that generates a 'soft gel' covering the ocular surface (in situ) is ideal for early forms of dry syndrome, while topical cyclosporine is the first new real prescription treatment for patients with moderate to severe forms of dry eye. Another potential agent to revolutionize the treatment of various disorders is based on the discovery of the purinergic receptor agonists. This is not only relevant for the production of mucin and the change in tear fluid content, but it may also have implications for other sinopulmonary disorders such as cystic fibrosis and chronic sinusitis. PMID- 15349043 TI - Nerve growth factor and the immune system: old and new concepts in the cross-talk between immune and resident cells during pathophysiological conditions. AB - PURPOSE OF REVIEW: This review provides an overview of nerve growth factor and its involvement in the immune system. Furthermore, recent data are provided revealing new important insights into the mechanisms of action of nerve growth factor in allergic reaction. RECENT FINDINGS: Recent studies on the effects of nerve growth factor on the immune cells involved allergic reaction, and on the potential role of nerve growth factor in tissue remodelling are presented. SUMMARY: Nerve growth factor has an extended function from the nervous system to a wide range of activities in the immune system. Several papers have highlighted the role of the factor in allergic inflammation. This review describes old and new concepts of nerve growth factor in the immune system: the relation between nerve growth factor and the main cells taking part in allergic inflammatory disorders, structural cells, mediators and cytokines/chemokines, as well as the mechanisms leading to nerve growth factor increase. Understanding these complex mechanisms will introduce new therapeutic approaches for nerve growth factor in the immune system, in addition to those already established in the nervous system. PMID- 15349044 TI - Developments in ocular allergy. AB - PURPOSE OF REVIEW: The goal of this article is to evaluate developments in the knowledge of inflammatory mechanisms and treatments of ocular allergy. RECENT FINDINGS: Research developments in ocular allergy summarized in this article include the following findings: (1) ocular epithelial cells play a role in inflammation; (2) respiratory syncytial virus is a pathogen in allergic conjunctivitis; (3) transglutaminase inhibitors reverse allergic related inflammation; (4) eosinophils and neutrophils both play a role in ocular allergy; (5) eotaxin-1 and eotaxin-2 play a role in eosinophilic recruitment; and (6) loteprednol etabonate, desonide phosphate, and cyclosporine have been shown to be effective and safe in the treatment of ocular allergy. SUMMARY: This article summarizes the research conducted for each of the above recent findings and outlines their clinical applications. PMID- 15349045 TI - Ocular cicatricial pemphigoid review. AB - PURPOSE OF REVIEW: Ocular mucous membrane pemphigoid produces progressive cicatrizing conjunctivitis; this can result in scarring of the conjunctiva and cornea. RECENT FINDINGS: Increased expression of macrophage-colony-stimulating factor, collagen-binding heat shock protein 47, transforming growth factor-beta1, IL-4, IL-5, and macrophage migration inhibitory factor may enhance both conjunctival inflammation and conjunctival scarring. Recent developments on mucous membrane pemphigoid medical therapy include the efficacious effect of daclizumab, intravenous immunoglobulin therapy, and methotrexate. Subconjunctival mitomycin has proved not to be efficacious in controlling long-term, conjunctival inflammation and scarring. The Boston scleral lens enhances vision, reduces the disabling ocular pain and photophobia, and helps to heal persistent epithelial defects, reducing recurrence of defects. Recent developments on mucous membrane pemphigoid surgical therapy include keratolimbal allografts and amniotic membrane transplantation, with or without penetrating keratoplasty for ocular surface reconstruction in total stem cell deficiency. The prognosis is strongly influenced by preoperative conditions such as tear function and functional external ocular adnexae, and by postoperative conditions such as persistent inflammation, severe dry eye, or rejection of the keratolimbal allograft. Some authors find efficacious the use of amniotic membrane transplantation for reconstruction of the conjunctival fornices provided systemic immunosuppression is pre- and post-operatively used. SUMMARY: The use of daclizumab or intravenous immunoglobulin therapy, the better selection of candidates for surgical interventions, and the better pre- and post-operative management of keratolimbal allograft and amniotic membrane transplantation may improve visual rehabilitation in the patients with ocular mucous membrane pemphigoid. PMID- 15349046 TI - Tear cytokines in acute and chronic ocular allergic inflammation. AB - PURPOSE OF REVIEW: Elevated levels of inflammatory cytokines have been reported in tears from ocular allergic disease states. The purpose of this review is to assimilate recent research contrasting tear cytokine concentrations in non allergic subjects versus subjects with acute (seasonal allergic conjunctivitis) and chronic (giant papillary conjunctivitis, vernal keratoconjunctivitis, atopic keratoconjunctivitis) ocular allergic inflammation to discover whether the cytokine profiles could provide useful insight into disease mechanisms and therapeutic targets. RECENT FINDINGS: Recent studies have revealed distinct differences in the cytokine/chemokine concentrations in tears between the various manifestations of ocular allergy. The acute (seasonal allergic conjunctivitis) and iatrogenic (giant papillary conjunctivitis) forms of ocular allergic inflammation are characterized by an overall lack of significant cytokine changes in tears compared with chronic disease (vernal keratoconjunctivitis, atopic keratoconjunctivitis). Chronic ocular allergic inflammation produces increased concentrations of T helper 1 and 2, and proinflammatory cytokines as well as chemokines. However, vernal and atopic keratoconjunctivitis portray distinct differences in the patterns of tear cytokines/chemokines expressed. SUMMARY: The plethora of increased cytokines and chemokines in vernal and atopic keratoconjunctivitis compared with non-allergic, seasonal allergic conjunctivitis and giant papillary conjunctivitis provides a new perspective into the complex inflammatory processes occurring on the ocular surface in chronic disease. The ability to measure multiple cytokines in tears, combined with knowledge obtained from in-vitro analysis of the individual and combined effects of these cytokines on various conjunctival cells (i.e. mast cells, epithelial cells, fibroblasts) has facilitated further understanding of specific processes contributing to maintenance of inflammation and progression of vision-threatening disease and paved the way toward new therapeutic targets. PMID- 15349047 TI - Objective measurement of allergic reactions in the eye. AB - PURPOSE OF REVIEW: Conjunctival allergen challenge has become a useful model of human ocular allergy and an accepted method for studying the effects of topical antiinflammatory medications. Conjunctival allergen challenge reactions are typically evaluated in a subjective fashion. Erythema and edema are graded by observation, using a 0 to 4 scale. Itching is graded on a 0 to 4 scale by questioning the subject. We have reviewed the recent literature on conjunctival allergen challenge, including recently developed methods aimed at objective assessment of ocular allergic reactions, such as the erythema, edema, sensation, or EES, method. RECENT FINDINGS: Conjunctival erythema can be measured objectively using a spectroradiometer, or colorimeter, a device which measures the chromaticity of reflected light, and provides the x, or u', and y, or v', coordinates of any color. Edema of the eyelids and conjunctiva can be measured using a fractional millimeter reticule in the eyepiece of a slit lamp microscope. Ocular surface sensation can be evaluated using the aesthesiometer of Cochet and Bonnet, or by using a 'sensation index', a measurement that takes into account the intensity and duration of itching. SUMMARY: CAC reactions can be measured objectively using the erythema, edema, sensation method, as well as subjectively by observation and questioning. PMID- 15349048 TI - Ocular allergic disease. AB - PURPOSE OF REVIEW: This review will focus on recent advances in our understanding of the pathogenesis of allergic eye diseases. Common findings in acute allergic conjunctivitis (seasonal and perennial) and chronic allergic conjunctivitis (vernal keratoconjunctivitis, atopic keratoconjunctivitis, and giant papillary conjunctivitis) include evidence of mast cell activation and eosinophil attraction and activation. Cytokine levels found in tears, conjunctival impression cytology and biopsy specimens, and serum have been evaluated as markers of disease, and as targets of therapeutic intervention. RECENT FINDINGS: Human conjunctival epithelial cells respond to tumor necrosis factor alpha, interleukin-1 beta, and interferon-gamma individually and in combination. Intracellular adhesion molecule-1 expression is upregulated by interleukin-1 beta and tumor necrosis factor alpha. Conjunctival epithelial cells release interleukin-8 in response to interleukin-1 beta and tumor necrosis factor alpha but not interferon-gamma. Supernatants from activated mast cells cause increased adhesion of eosinophils to conjunctival epithelium. Tear levels of tumor necrosis factor alpha were elevated in vernal keratoconjunctivitis patients compared with normal controls. T cell lines from chronic allergic eye disease patients showed inconsistent production of cytokines in atopic and vernal keratoconjunctivitis and low levels in giant papillary conjunctivitis. Vernal keratoconjunctivitis patients have differing levels of eosinophil cationic protein in their serum if they were serum specific immunoglobulin E positive compared to serum specific immunoglobulin E negative patients. SUMMARY: Recent findings continue to expand our basic knowledge of mechanisms and differences between seasonal and perennial allergic conjunctivitis and atopic and vernal keratoconjunctivitis. Understanding the complex interactions and cross talk between cells, cytokines and other mediators is relevant for new therapeutic approaches directed at specific disease entities. PMID- 15349051 TI - Reaching for recognition, Magnet style. PMID- 15349053 TI - R-E-S-P-E-C-T, find out what it means to you. AB - : Use proper etiquette during confrontational situations. PMID- 15349054 TI - Partnering for better patient care. PMID- 15349055 TI - Dare to shift cultural behaviors. PMID- 15349056 TI - Know your options with the periodic performance review process. AB - Learn how Shared Visions-New Pathways synchs with the periodic performance review process. PMID- 15349057 TI - Inside the Nurse Practice Act. AB - : Nursing practice statutes provide patient care guidelines for each state. PMID- 15349061 TI - Making the communication connection. AB - Many of healthcare's greatest challenges involve miscommunication. Here, consider nursing-specific tips and tools for truly reaching all involved parties. PMID- 15349062 TI - An invitation to Magnet accreditation. AB - Review Magnet program basics and eligibility requirements to determine your organization's readiness for undergoing certification. PMID- 15349064 TI - Applied technology rounds out e-documentation. AB - Highlights of an electronic healthcare record include clinical reminders, progress note templates, and a clinical reference system. PMID- 15349065 TI - High-tech, high-touch: why wait? AB - Heighten your awareness of untapped information technology resources. PMID- 15349066 TI - Better by design: self-learning cardiac curricula. AB - Clinical specialists at Wausau Hospital, Wisc., devise continuing-education opportunities for cardiac care providers. PMID- 15349067 TI - Keep your eyes on defined contribution plans. AB - Anxious to find alternatives to an established system of employment-based health insurance, employers are increasingly investigating defined contribution models. PMID- 15349068 TI - The Magnetic pull: does your facility have what it takes to attain Magnet recognition? AB - Review the processes and unique offerings through which six hospitals gained the coveted Magnet recognition from the American Nurses Credentialing Center. PMID- 15349069 TI - Move care to a higher level with emergency systems. AB - Consider using emergency department information systems to improve patient care in your facility. PMID- 15349071 TI - Share your vision, voice. AB - Use the approach discussed here to develop sharing relationships between nurse executives and bedside caregivers. PMID- 15349077 TI - What does freedom look like? PMID- 15349078 TI - Environment- and gene-dependent human ontogenesis, sociogenesis and phylogenesis (eco-geno-onto-socio-phylogenesis). AB - Prevention of environment- and gene-dependent, teratogenic malfunctions ("Functional Teratogenesis")-- caused by abnormal hormone, neurotransmitter and cytokine concentrations during organization of the neuro-endocrine-immune system (NEIS) should be considered as a global challenge of outstanding relevance. By optimizing the natural and social environment and correcting in time abnormal concentrations of hormones, neurotransmitters and cytokines during the critical perinatal (pre- and early postnatal) organization period of the NEIS ("Neuro Endocrine-Immune Prophylaxis") human ontogenesis and sociogenesis can be decisively improved ("Primary Prevention of Maldevelopments of Human Beings and their Societies"). Finally, phylogenesis is dependent on incessant sequencies of ontogenesis and sociogenesis ("Onto-Socio-Phylogenesis"). PMID- 15349079 TI - Periodic emergence of great poets in the history of Arabia & Persia, China and Japan. AB - A periodicity of about 500 years has been discovered in the history of poetry and documented by means of inferential statistics. Great poets of Arabia, Persia, China and Japan emerged periodically every 500 years. Moreover, the waves of poetic creativeness in the West and in the East have been synchronous for the last 3000 years. It is a surprising fact, that this periodicity has been known already before 800 B.C. to the priests of Babylon, who ascribed it to the influence of goddess Inanna. A set of psychological symptoms typical for pubescence recurs on a global (worldwide) scale during these historical epochs every 500 years. One possible explanation would be to search for a cosmophysical factor, which impacts the neuroendocrine system of men. PMID- 15349080 TI - Precocious puberty associated with a pineal cyst: is it disinhibition of the hypothalamic-pituitary axis? AB - Accelerated development of secondary sexual characteristics or sexual precocity is a well-known entity. Most authors recognize two groups of patients, those described as having central precocious puberty (CPP) and those with precocious pseudopuberty. CPP results from premature activation of the hypothalamic pituitary-gonadal axis and pseudopuberty is caused by lesions that secrete gonadotropin-like substances or hormones. The onset of CPP is usually before age 8 in females and age 9 in males; however, there is contention that the age of onset is much earlier and also differs depending on the patients' race. Previously reported causes of CPP include intracranial neoplasm, infection, trauma, hydrocephalus and Angelman's syndrome. Pineal cysts are usually asymptomatic incidental findings, but have been associated with CPP. We present an interesting case of a patient with CPP and an associated pineal cyst. We review the literature on the pathogenesis of CPP and associated pineal cyst, the neuroendocrine relationship between the pineal gland and puberty and the neurosurgical role in these cases. PMID- 15349081 TI - Vitamin B 12 plasma concentrations in Alzheimer disease. AB - In this contribution we investigated the correlation between plasma vitamin B12 levels and cognitive impairment in Alzheimer's disease. We could demonstrate a significant inverse correlation when the MMSE scores of those patients with the lowest 10% Vitamin B12 plasma levels (<184 ng/ml) were compared with the upper 10% Vitamin B12 plasma levels (>598 ng/ml): p=0.008, Spearman-Rho= -0.36. MMSE in the upper percentile of plasma B12 levels was 20.0 +/- 4.6 and in the lower percentile 15.7 +/- 6.1, resulting in a difference of 4.3 MMSE points. We conclude, that vitamin B12 deficiency could aggravate or accelerate the course of Alzheimer disease as vitamin B12 possesses neuroprotective and anti-inflammatory properties. PMID- 15349082 TI - The effect of Vigabatrin, Lamotrigine and Gabapentin on the fertility, weights, sex hormones and biochemical profiles of male rats. AB - PURPOSE: A case control study was conducted to assess the effect of Sabril (Vigabatrin), Lamictal (Lamotrigine) and Neurontin (Gabapentin) on fertility in male rats. Their effect on the body and organs weight and certain biochemical profiles including total serum protein, cholesterol, triglycerides, serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), serum testosterone, and FSH levels were also measured. METHODS: several parameters, concerning fertility were measured in 40 albino male rats of Sprague Dawley strain, they were divided into 4 groups, group one received vehicle (distilled water), group two received Vigabatrin in a dose of 200 mg/kg body weight, group three received Lamotrigine in a dose of 30 mg/kg body weight, and group four received Gabapentin 100 mg/kg body weight. All the male rats in these groups received the different medications for a complete reproductive cycle (60 days). After 24 hours of the last dose, the animals were weighed and autopsied under light ether anesthesia. Parameter of fertility that has been measured in this study includes: sperm count and motility, weight of different reproductive organs, germ cell and interstitial cell population, serum testosterone and FSH levels and assessment of pregnancies in females mixed with tested males. Biochemical profiles such as serum cholesterol, serum triglycerides, serum bilirubin, SGOT, SGPT level are all measured. The results of the histological, histometerical studies and biochemical profiles were compared to that of the control group, and the significance of these results was measured using student's "t" test. RESULTS: There was significant reduction in the body weight and the weight of the testes, epididymis, seminal vesicles, ventral prostate, and vas deferens in the antiepileptic fed male rats in comparison to the control group (p > 0.001). There was significant reduction in testicular cells population dynamics including both germinal cell types and interstitial cell types in the antiepileptic fed male rats in comparison to the control group. There was also significant reduction in histometrical parameters and sperm dynamics in the antiepileptic fed male rats histologies in comparison to the control group. There was significant reduction in both testosterone and FSH levels (p < 0.001) in the antiepileptics fed male rats in comparison to the control group. There was also significant reduction in pregnancy rate observed in female rats exposed to the tested male rats among antiepileptic fed male rats compared to controls. The results of biochemical profiles assessment showed significant reduction in serum glucose, serum cholesterol, serum triglycerides levels and significant increase in serum bilirubin, SGOT, and SGPT levels in antiepileptics fed male rats in comparison to the control group. CONCLUSIONS: Fertility rate and other parameters concerned with fertility, sex hormones and certain biochemical profiles were significantly disturbed in male rats fed with three of the second-generation antiepileptic drugs Vigabatrin, Lamotrigine, and Gabapentin, indicating a possible toxic effect of these three medications on sexual organs, liver, and lipid metabolism. PMID- 15349083 TI - Nitric oxide modulates the physiological control of ciliary activity in the marine mussel Mytilus edulis via morphine: novel mu opiate receptor splice variants. AB - OBJECTIVES: The study sought to determine how dopamine controls ganglionic processes involved with modulating lateral cilia beating via the peripheral branchial nerve. METHODS: The lateral cilia found on the gill filaments exhibit metachronal ciliary beating determined stroboscopically. Novel opiate receptors were determined pharmacologically and demonstrated by RT-PCR and sequence analysis of total RNA from Mytilus edulis visceral ganglia. RESULTS: Dopamine applied to the visceral ganglion inhibits the activity of lateral cilia in a concentration and haloperidol sensitive manner. Morphine or DAMGO significantly enhances ciliary beating in a naloxone sensitive manner, whereas L-NAME, a nitric oxide synthase inhibitor, only antagonized morphine's action. SNAP, a nitric oxide donor, also enhanced lateral ciliary beating rates. Supporting the observation, i.e., morphine sensitive nitric oxide enhancement of ciliary beating and DAMGO insensitive, that two different mu opiate receptors are present in this tissue, a 602 bp fragment of the human micro 3 opiate receptor and a 935 bp fragment, designated micro 4 have been demonstrated. CONCLUSIONS: The lateral epithelium of the gill is innervated by serotonergic, cilioexcitatory neurons and dopaminergic, cilioinhibitory neurons, originating in the visceral ganglion. This data supports previous reports that demonstrate inhibiting ganglionic dopamine release allows the serotonin signals to prevail uncontrolled, enhancing ciliary rates. Supporting the observation that two different mu opiate receptors are present in this tissue, evidence is presented that identifies a 602 bp fragment of the human micro 3 opiate receptor and a 935 bp fragment, designated micro 4. Overall, the data strongly suggests that the two alternatively spliced mu opiate receptors may be involved in the physiological regulation of lateral ciliary activity in the visceral ganglia via dopamine and nitric oxide. PMID- 15349084 TI - Leptin gene expression in the hypothalamus and pituitary of pregnant pigs. AB - BACKGROUND: Leptin, the 16-kDa peptide hormone product of the ob gene, is a regulatory hormone secreted mainly by adipose tissue. Recent studies have shown leptin production by other tissues, including rat hypothalamus, rat and human pituitary, rat skeletal muscle, kidney and stomach, human and porcine placenta, human mammary epithelial cells as well as endometrial tissues. This hormone is a central modulator of food intake, metabolism and neuroendocrine functions. OBJECTIVES: The aim of the study was to detect and locate porcine leptin gene expression in the different areas of the hypothalamus and pituitary on days 14-16 and 30-32 of pregnancy in pigs. METHOD: Leptin gene expression was analysed by RT PCR method. PCR products were subjected to sequencing analysis. RESULTS: Leptin mRNA was expressed in the medial basal hypothalamus, preoptic area, stalk median eminence, anterior pituitary, posterior pituitary and adipose tissue on days 14 16 and 30-32 of pregnancy. Sequence analysis of the 258 bp product from the hypothalamus and pituitary confirmed 99% homology with the corresponding region of porcine leptin cDNA sequence. CONCLUSION: Leptin mRNA expression in the porcine hypothalamus and pituitary gland implies its paracrine and/or autocrine role in the regulation of hypothalamic-pituitary axis activity. PMID- 15349085 TI - Serum testosterone and corticosterone levels in acute experimental autoimmune encephalomyelitis (EAE) in male Wistar rats. AB - OBJECTIVES: Experimental autoimmune encephalomyelitis (EAE) was induced to investigate the levels of circulating total testosterone (TT) and the possible association of corticosterone with the steroid-producing capacity of the testes. SETTING AND DESIGN: We determined gonad weights, serum TT and corticosterone levels during the development of EAE in male rats. METHODS: Active EAE was induced in young male Wistar rats by injection of whole myelin in complete Freund's adjuvant. All the rats were weighted and monitored daily for clinical signs and blooded during different phases of the disease. Serum TT was measured by radioimmunoassay and circulating corticosterone was determined using a competitive enzyme immunoassay. Also testes and seminal vesicles were removed to determine their weights. RESULTS: Seminal vesicle weights and serum TT levels diminished during the acute stage of the disease in all EAE rats and then they began gradually to increase, reaching the normal values at the post-recovery phase. Concomitantly a significant increase in serum corticosterone levels was observed during the acute EAE and the post-recovery phase was accompanied by a decline in corticosterone levels. MAIN FINDING: Our results indicated an inverse correlation between serum TT and corticosterone levels during the acute EAE. Moreover, the diminution of TT was not a consequence of an alteration of the testes induced by anti-myelin antibodies neither contributed by apoptosis of testis cells by exposure to corticosterone. CONCLUSIONS: The negative correlation between corticosterone and TT levels associated to EAE may be relevant in understanding the association of the endocrine system and the development of autoimmune demyelinating diseases. PMID- 15349086 TI - The influence of chronic exposure to low frequency pulsating magnetic fields on concentrations of FSH, LH, prolactin, testosterone and estradiol in men with back pain. AB - OBJECTIVES: There is widespread public concern that electromagnetic fields might be hazardous. However, studies on the biological effects of magnetic fields (MFs) have not always been consistent. Influence of extremely-low frequency MFs used in physiotherapy on endocrine system was rarely examined. Therefore, the aim of the present study was to investigate the concentrations of some pituitary (FSH, LH, prolactin) and sex (testosterone, estradiol) hormones in men with back pain exposed to magnetic fields applied during magnetotherapy or magnetostimulation over the period of three weeks. MATERIAL AND METHODS: The study was performed on 20 men aged 28-62 years (mean+/-SEM: 46.4+/-2.0 years) suffering from chronic low back pain who underwent magnetotherapy (10 patients, mean age+/-SEM: 48.4 years, range: 28-62 years) or subjected to magnetostimulation (10 patients, mean age+/ SEM: 44.3 years, range: 34-52 years) for 15 days (daily at 10:00 h, with weekend breaks). Blood samples were collected at 08:00 before magnetic field application, one day and one month following the application. Concentrations of hormones were measured by micromethod of chemiluminescence. RESULTS: Both magnetotherapy and magnetostimulation lowered levels of prolactin. The levels of LH decreased significantly one month after magnetotherapy in comparison with the baseline whereas following magnetostimulation slight but insignificant increase was observed. Estradiol concentrations were significantly lower one day and one month following magnetosimulation in comparison to the baseline and did not change after magnetotherapy. No statistically significant changes were observed in levels of FSH and testosterone after either magnetotherapy or magnetosimulation at any time examined. CONCLUSION: Magnetic fields applied in physiotherapy exert no or very subtle effect on concentrations of FSH, LH, prolactin, testosterone, and estradiol in men. PMID- 15349087 TI - Increased mitogen-induced lymphocyte proliferation in treatment resistant depression: a preliminary study. AB - BACKGROUND: The biological basis of treatment resistance in depression still remains unclear. Some studies suggest that there exists a relationship between a disturbance in the immune system and depression. AIMS: To investigate the immune status of patients with treatment resistant depression, we compared the in-vitro lymphocytic responses to three different mitogens in 10 drug-free patients with treatment resistant depression and 10 normal controls matched for age and sex. RESULTS: We observed a marked increase in the lymphocyte mitogenic activity to mitogens of T lymphocytes. DISCUSSION: This activation of lymphocyte function may be indicative of a pathological stimulation in cell-mediated immunity in patients with treatment resistant depression. PMID- 15349088 TI - The beneficial effect of amalgam replacement on health in patients with autoimmunity. AB - BACKGROUND: Patients with certain autoimmune and allergic diseases, such as systemic lupus, multiple sclerosis, autoimmune thyroiditis or atopic eczema, often show increased lymphocyte stimulation by low doses of inorganic mercury in vitro. The patients often report clinical metal hypersensitivity, especially to nickel. OBJECTIVE AND METHODS: In this study we examined the health impact of amalgam replacement in mercury-allergic patients with autoimmunity. The suitability of MELISA, an optimized lymphocyte stimulation test, for the selection of susceptible patients and monitoring of sensitization was also examined. Amalgam fillings were replaced with composites and ceramic materials. Follow-up health status and lymphocyte reactivity were assessed and evaluated half a year or later following amalgam removal. RESULTS: Results of lymphocyte reactivity measured with MELISA indicate that in vitro reactivity after the replacement of dental amalgam decreased significantly to inorganic mercury, silver, organic mercury and lead. Out of 35 patients, 25 patients (71%) showed improvement of health. The remaining patients exhibited either unchanged health (6 patients, 17%) or worsening of symptoms (4 patients, 11%). The highest rate of improvement was observed in patients with multiple sclerosis, the lowest rate was noted in patients with eczema. The initial mercury-specific lymphocyte reactivity was significantly higher in the responder group, than in the non-responders, whose health was not improved by amalgam removal. All patients with health improvement after amalgam replacement showed reduced proliferation to inorganic mercury in follow-up MELISA. In vitro responses to phenylmercury and nickel did not differ between the groups. CONCLUSIONS: Mercury-containing amalgam may be an important risk factor for patients with autoimmune diseases. MELISA is a valuable tool for selection of patients for amalgam replacement and also for monitoring of metal allergies. PMID- 15349089 TI - Hypothalamic nitric oxide synthase activity during sexual maturation in female rats. Effects of estrogens. AB - OBJECTIVES: To determine the hypothalamic activity of nitric oxide synthase (NOS, the enzyme involved in the synthesis of nitric oxide NO) during sexual maturation in prepubertal (15 days old) and peripubertal female rats (30 days old) as well as the effect of estradiol administration on this neurotransmitter system. METHODS: Hypothalamic samples containing the anterior preoptic and medial basal areas (APOA-MBH) were homogenized with HEPES 20 mM, pH = 7.4 and NOS activity was determined in APO-MBH after 10 minutes of incubation by the conversion of 14C arginine to 14C citrulline. RESULTS: The hypothalamic concentration of NOS is significantly higher in peripubertal than in prepubertal rats. Treatment with EB increased significantly the activity of the enzyme in both groups compared with control and the increases was similar at both ages. CONCLUSIONS: These results clearly demonstrated that the hypothalamic NOS activity increases in peripubertal rats as compared with prepubertal animals. Estradiol has a similar stimulatory effect on hypothalamic NOS activity at both ages of sexual maturation, indicating that the increase in NOS during sexual maturation is connected with the peripubertal increase of estradiol rather than an increase in the sensitivity of the enzyme to the ovarian hormone. PMID- 15349090 TI - Effect of bromocriptine on cell apoptosis and proliferation in GH3 cell culture. AB - OBJECTIVES: In our study, with the use of GH3 cells line we decided to examine 1) what is the relation between the dose of bromocriptine and the development of apoptosis in GH3 cells 2) whether the induction of apoptosis is accompanied by alterations in bcl-2 and p53 content and 3) whether dibutyryl-cAMP or phorbol esters affect the initiation of apoptosis in GH3 cells. RESULTS: The current study demonstrated the absence of alterations in GH3 cells incubated for 24 h with bromocriptine at the concentrations of up to 15 micromol/l. Apoptotic and necrotic changes were observed after 48 h incubation with bromocriptine at the concentrations of 25 micromol/l. The ratio of necrotic to apoptotic cells increased at 40 micromol/l of bromocriptine concentration. An inhibitory effect of bromocriptine on cell proliferation was also observed. Phorbol-12-myristate-13 acetate (PMA), at concentrations ranging between 25 ng to 200 ng/ml, reduced the amount of apoptotic cells. CONCLUSIONS: Application of dibutyryl-cAMP at the concentration of 1 to 8 mmol/l resulted in an inhibition of apoptosis, followed by an increase in the number of cultured cells. Ultrastructural studies showed evident apoptotic lesions in the cells. PMID- 15349091 TI - An overview of genomic data analysis. PMID- 15349092 TI - DNA microarrays and data analysis: an overview. PMID- 15349093 TI - Tracking gene-disease relationships for high-throughput functional studies. PMID- 15349094 TI - Genomic approaches in cancer biology. PMID- 15349095 TI - Application of microarray profiling to clinical trials in cancer. PMID- 15349097 TI - Fluorescence spectroscopy accurately detects irreversible cell damage during hepatic radiofrequency ablation. AB - BACKGROUND: A current limitation of hepatic radiofrequency ablation (RFA) is an inability to detect ablation margins in real time. Thermal injury from RFA alters the biochemical properties governing tissue fluorescence. We hypothesized that the changes in hepatic fluorescence measured during hepatic RFA could be used to detect irreversible hepatocyte damage accurately and to determine ablation margins in real time. METHODS: RFA was performed on healthy pig livers and monitored in vivo simultaneously for fluorescence and temperature by a fiberoptic micro-interrogation probe connected to a spectroscopy system. Ablations were stopped based on previously established real-time fluorescence spectral data, not based on temperature or time. To determine where in the ablated tissue cell death occurred, biopsies for transmission electron microscopy were taken from 4 areas of 3 specimens: (1) nonablated liver, (2) hemorrhagic zone/normal liver interface, (3) hemorrhagic zone/coagulated zone interface, and (4) coagulated zone. In vitro fluorescence emission intensity was determined at each biopsy site. RESULTS: Peak hepatic fluorescence intensity occurred at 470 nm and decreased as RFA progressed. Transmission electron microscopy evidence of irreversible hepatocyte damage occurred at the interface of the coagulation zone and the hemorrhagic zone and correlated with a 87.5% +/- 9% decrease in fluorescence emission intensity. Tissue fluorescent changes from thermal injury were unaffected by tissue cooling. CONCLUSION: Fluorescence spectroscopy accurately detected hepatocellular thermal injury from RFA in real time and can detect irreversible cell damage during tissue thermal therapy. PMID- 15349099 TI - Regulatory cells develop after the spontaneous acceptance of rat liver allografts. AB - BACKGROUND: After donor-specific transfusion, tolerance to heart transplants is serially passed to naive rats by the adoptive transfer of long-term survivor (LTS)-tolerant splenocytes (SC). We examined whether regulatory cells similarly develop after the spontaneously accepted Lewis (LEW) to Dark Agouti (DA) liver transplants. METHODS: SC from a LTS DA rat with a LEW liver were adoptively transferred to a naive DA 1 day before transplantation of an irradiated (1000 rad) LEW liver. RESULTS: Untreated LEW to DA liver allografts were uniformly accepted; whereas all irradiated LEW liver grafts were rejected. In contrast, when 1.5 x 10 8 DA LTS SC were transferred to a naive DA recipient, all irradiated LEW liver grafts were accepted. When decreased to 1.0 x 10 8 LTS DA SC, only 1 of 4 irradiated LEW grafts was accepted. However, if 1.5 x 10 8 DA SC harvested only 30 days after liver transplantation were transferred, only 2 of 5 irradiated LEW liver grafts were accepted. The serial second and third adoptive transfers of 1.5 x 10 8 DA LTS SC also resulted in the uniform acceptance of irradiated LEW livers. CONCLUSION: Regulatory cells that develop after the spontaneous acceptance of a LEW to DA liver transplant can serially transfer tolerance to new naive LEW liver allograft DA recipients. This "infectious tolerance" is dependent on the time of cell harvest after transplantation and on the cell dose given. PMID- 15349100 TI - A novel approach to xenotransplantation combining surface engineering and genetic modification of isolated adult porcine islets. AB - BACKGROUND: Effective cytoprotection to xenoislets would circumvent the major tissue limitation for pancreatic islet transplantation (PIT). Cell-surface engineering with poly[ethylene glycol] (PEG) derivatives can successfully prevent antibody binding to the surface antigens. Gene transfer of the antiapoptotic Bcl 2 gene has been shown to decrease cytotoxicity mediated by xenoreactive natural antibodies and complement. In this study, we assessed survival and function of surface-engineered porcine islets genetically modified to overexpress Bcl-2. METHODS: Incorporation of PEG derivatives into the islet surface and adenovirus mediated gene transfer of Bcl-2 (AdBcl-2) was accomplished within 24 hours post isolation. Cytotoxicity induced by human xenoreactive natural antibodies was evaluated by islet intracellular lactate dehydrogenase release and microscopic analysis using membrane-integrity staining. Islet functionality was assessed by static incubation and after intraportal infusion (5000 IEQ) into diabetic NOD SCID mice reconstituted with human lymphocytes (5 x 10 8 /intraperitoneally/15 days before PIT). RESULTS: No significant change in islet viability, morphology, and functionality was demonstrated after the incorporation of PEG-mono-succimidyl succinate (MSPEG), or PEG-di-succimidyl-succinate "end"-capped with albumin (DSPEG) with or without gene transfer of Bcl-2. Islets treated with MSPEG presented a significant reduction in lactate dehydrogenase release compared with controls (41.2 +/- 3 vs 72.1 +/- 7, respectively, P <.05). Further protection was accomplished by DSPEG or AdBcl-2. The maximal cytoprotection was achieved by DSPEG +AdBcl-2 (15.5 +/- 4.9%, P <.001). Nonfasting glucose >200 mg/dL was found in 100% of the animals given control islets (n = 6) within 48 hours post transplant. In contrast, euglycemia was achieved in 100% of the animals given islets modified with DSPEG + AdBcl-2 during the observation time. CONCLUSIONS: Surface-engineering with functionalized PEG derivatives in combination with genetic modification with Bcl-2 significantly reduced islet loss after PIT. Application of this novel technology may improve results in xenoislet transplantation. PMID- 15349101 TI - Glucose transporter-1 gene expression is associated with pancreatic cancer invasiveness and MMP-2 activity. AB - BACKGROUND: Overexpression of the facilitative glucose transporter-1 (GLUT-1) has been observed for a wide range of human cancers, with the degree of overexpression generally being inversely correlated with prognosis. We tested the effects of modulating GLUT-1 expression on pancreatic cancer cellular invasiveness. METHODS: GLUT-1 expression in MIAPaCa-2, PANC-1, BXPC-3, and CAPAN 2 cells was assayed using Western blotting. Cells were stably transfected with a GLUT-1 expression vector or a GLUT-1 RNA interference vector to alter GLUT-1 expression. Matrix metalloproteinase-2 (MMP-2) activity and expression were assayed using zymography and Western blotting, respectively. In vitro cellular invasiveness was assayed using Matrigel Boyden chambers, and in vivo metastatic potential was assessed using a nude mouse xenograft model. RESULTS: Variable baseline GLUT-1 expression levels were detected among the cell lines. Forced overexpression of GLUT-1 induced increases in MMP-2 expression and activity and in cellular invasiveness. GLUT-1 silencing induced reductions in MMP-2 expression and activity, cellular invasiveness, and metastatic potential in vivo. CONCLUSION: GLUT-1 promotes pancreatic cellular invasiveness. The therapeutic implications of this finding warrant further study. PMID- 15349102 TI - The role of the glutathione antioxidant system in gut barrier failure in a rodent model of experimental necrotizing enterocolitis. AB - BACKGROUND: Evidence suggests that intestinal barrier failure in necrotizing enterocolitis results in part from overproduction of nitric oxide and other toxic oxidant species that result in enterocyte death and intestinal barrier failure. We hypothesize that the glutathione detoxifying system is important in maintaining intestinal barrier integrity by protecting against nitrosative stress. METHODS: Newborn rats were subjected to hypoxia (5% O2, tid) and fed formula by gavage (NEC), or were breast-fed without hypoxia (BF). Rats were killed and the distal ilea were harvested for RNA, protein, and morphologic studies. RNA underwent cDNA microarray analysis. To assess glutathione in protecting against nitrosative stress, IEC-6 cells were exposed to SIN-1 and/or L buthionine-(S,R)-sulfoximine (BSO), a glutathione inhibitor. Cells were analyzed for glutathione-S-transferase activity, apoptosis and mitochondrial function. RESULTS: BF controls developed normal intestinal architecture, whereas NEC animals sustained damage to the intestinal epithelium. Microarray analysis demonstrated that 93 genes were overexpressed in NEC compared with controls. In the array, glutathione-S-transferase pi and alpha 2, GSH-dependent detoxifying enzymes, RNA were upregulated compared with BF controls. IEC-6 cells exposed to SIN-1/BSO produced an increase in apoptosis. Poly ADP-ribosylpolymerase cleavage and apoptosis-inducing factor (AIF) nuclear localization, markers of apoptosis, were seen in IEC-6 cells exposed to SIN-1/BSO as opposed to media controls. CONCLUSION: These data support the hypothesis that GSH antioxidant system plays a crucial role in gut barrier protection by attenuating enterocyte death. PMID- 15349103 TI - Predictors for medical students entering a general surgery residency: National survey results. AB - BACKGROUND: The number of general surgery (GS) residency applicants had been decreasing before 2003. This national survey of fourth-year medical students elucidates factors related to the basic surgery clerkship that are associated with the decision to enter a GS residency. METHODS: A national sample of 2250 fourth-year medical students from all 4-year allopathic US medical schools was surveyed in spring 2002. Multivariate analyses were performed to identify mutable predictors for students entering GS. RESULTS: Data from 1531 fourth-year medical students from 121 different medical schools (response rate=68%) showed that 5.6% planned to enter GS. In multivariate analyses, the strongest predictor of entering GS was satisfaction with the quality of attending teaching (odds ratio 2.14, P <.01) in surgery clerkships. Several clerkship factors, such as frequency of call nights and total hours worked., were not as strongly associated with entering GS residency, Subsequent analyses showed that predictors of satisfaction with the quality of attending teaching included intraoperative activities (ie, suturing, cutting, and stapling), having attending-led rounds, and performing a history and physical with an attending. Significant negative predictors of satisfaction included observing or retracting only in surgery. CONCLUSIONS: In this national survey, factors are identified that are significantly associated with students entering a GS residency. Some of these mutable factors may increase the pool of GS residency applicants. PMID- 15349104 TI - Transducible recombinant small heat shock-related protein, HSP20, inhibits vasospasm and platelet aggregation. AB - BACKGROUND: Human saphenous vein (HSV) is the autologous conduit of choice for peripheral vascular reconstruction. Injury during harvest leads to vasospasm and a thrombogenic endoluminal surface. A proteomic transduction approach was developed to prevent vein graft vasospasm and thrombosis. METHODS: Recombinant HSP20 protein linked to the TAT protein transduction domain was generated in a bacterial expression system (TAT-HSP20). The effect of this protein on the inhibition of smooth muscle contraction was determined using rings of rabbit aorta and HSV in a muscle bath. In addition, the effects of TAT-HSP20 on platelet aggregation were determined in vitro using human citrated whole blood. RESULTS: Recombinant TAT-HSP20 inhibited norepinephrine-induced contraction of rabbit aortic and HSV segments. Similarly, TAT-HSP20 induced smooth muscle relaxation in HSV segments precontracted with norepinephrine. In human-citrated whole blood, platelet aggregation was significantly inhibited by TAT-HSP20 in a dose-dependent manner. CONCLUSIONS: The results of this study demonstrate that recombinant TAT HSP20 inhibits vascular smooth muscle contraction and platelet aggregation. This suggests that HSP20 may be an ideal effector molecule to target as a proteomic approach to enhance early vein graft patency rates by preventing acute vasospasm and thrombosis. PMID- 15349105 TI - Carcinoembryonic antigen directed herpes viral oncolysis improves selectivity and activity in colorectal cancer. AB - BACKGROUND: G207 is an oncolytic herpes virus whose replicative cycle requires cellular ribonucleotide reductase (RR) for viral DNA synthesis. We attempt to enhance viral cytotoxicity in carcinoembryonic antigen (CEA)-producing colorectal cancer (CRC) cells through CEA-driven RR production. METHODS: CEA enzyme-linked immunosorbent assay was performed on LS174T and HCT-8 human CRC cells. The CEA enhancer-promoter (CEA E-P) was functionally assessed by luciferase assay. CEA E P was cloned upstream of UL39, the gene encoding the large subunit of RR. Cells were transfected with CEA E-P/UL39 and infected with G207 for cytotoxicity assays. LS174T, with or without CEA E-P/UL39, were implanted into athymic mouse flanks (n = 28) and treated with G207. RESULTS: CEA levels were 7-fold higher in LS174T versus HCT-8 ( P <.00001). CEA E-P increased luciferase expression 7.5 fold in LS174T ( P <.01), with no increase in HCT-8. G207 cytotoxicity of'CEA E P/UL39-transfected LS174T cells increased 69% by day 10 versus nontransfected cells ( P <.001), with no significant increase in HCT-8. Combining CEA E-P/UL39 with G207 in LS174T flank tumors resulted in a 65% decrease in tumor volume versus G207, phosphate-buffered saline, or'CEA E-P/UL39 alone ( P <.0001). CONCLUSIONS: CEA-driven RR production by CEA-secreting CRC cells significantly improves oncolytic viral cytotoxicity and specificity in vitro, and reduces tumor burden in vivo. PMID- 15349106 TI - Double-gene ablation of SSTR1 and SSTR5 results in hyperinsulinemia and improved glucose tolerance in mice. AB - BACKGROUND: Previous studies conducted in our laboratory showed that single-gene ablation of somatostatin receptor (SSTR)1 or 5 results in diabetes in mice. The objective of this study was to determine the effect of double-gene ablation of SSTR1 and SSTR5 on insulin secretion and glucose homeostasis in mice. METHODS: SSTR1/5 -/- mice and wild-type (WT) control mice were generated and their genotype verified via polymerase chain reaction. Insulin secretion and glucose levels in these mice were examined with the use of an intraperitoneal glucose tolerance test (1.2-2.0 g/kg body weight). In vitro glucose-stimulated insulin secretion was studied with the use of the isolated perfused mouse pancreas model and islet culture techniques. Pancreata morphologic alterations were determined, and an immunohistochemistry analysis was performed. RESULTS: In vitro incubation of isolated islets from WT mice with somatostatin peptides resulted in significant reduction in insulin secretion, whereas SSTR1/5 -/- mouse islets had no response to somatostatin peptides confirming SSTR1/5 gene ablation. SSTR1/5 -/ mice also had significant increase of both basal and glucose-stimulated insulin levels in vitro. During the intraperitoneal glucose tolerance test, SSTR1/5 -/- mice had significantly improved glucose tolerance and sustained an increase in late-phase insulin secretion in vivo. Histological analysis demonstrated significant islet hyperplasia in the SSTR 1/5 -/- mouse pancreas. Immunostaining revealed an overall increase of glucagon and pancreatic polypeptide-producing cells in the islets of SSTR1/5 -/- mice. CONCLUSIONS: Double-gene ablation of SSTR1 and SSTR5 in mice resulted in a distinct phenotype with islet cell hyperplasia, hyperinsulinemia, and improved glucose tolerance. This form of diabetes differs from that seen in mice in which only the SSTR1 or SSTR5 gene was ablated. These results demonstrate that SSTR1 and SSTR5 are important regulators of insulin secretion and glucose regulation, and suggest that SSTR1 and SSTR5 are coordinately regulated. PMID- 15349107 TI - A prospective, randomized, controlled clinical trial of tissue adhesive (2 octylcyanoacrylate) versus standard wound closure in breast surgery. AB - BACKGROUND: Recent studies suggest that the use of tissue adhesive for closure of both traumatic lacerations and incisional surgical wounds leads to cosmetic outcome comparable to conventional sutures. To date, no studies have investigated tissue adhesive in breast surgery and costs. Our aim was to compare the tissue adhesive 2-octylcyanoacrylate (OCA) with standard suture in breast surgery. METHODS: A prospective randomized study was conducted in which 151 patients were assessed for eligibility, and 133 were randomly allocated to skin closure with OCA adhesive or monofilament suture. Cosmetic outcome with blinded assessment, wound management by the patients, complication rates, and economic outcome were recorded. RESULTS: There was no difference in cosmetic score in the 2 groups, nor in complications at the early, 6-month, and 1-year follow-up. Patient satisfaction with the wound closed with OCA was rated significantly higher when compared with standard suture (P <.0001). The application of the tissue adhesive was significantly faster than that for standard suture (P <.001). In economic terms total costs were less in the tissue adhesive group, mainly due to lower postoperative costs of physician and assistant services (P <.001). CONCLUSIONS: OCA is effective and reliable in skin closure for breast surgery, yielding similar cosmetic results to standard suture. OCA is faster than standard wound closure and offers several practical advantages over suture repair for patients. Cost analysis has found that OCA adhesive can significantly decrease health care costs. PMID- 15349108 TI - Influence of opioid use on surgical and long-term outcome after resection for chronic pancreatitis. AB - BACKGROUND: The outcome of pancreatic resection for chronic pancreatitis in patients with preoperative opioid use is not well described. METHODS: During 1997 to 2003, 112 of 231 patients referred with chronic pancreatitis underwent pancreatic resection. The outcome of patients who had preoperative opioid use (N=46) was compared with those without (N=66). RESULTS: Patients who used opioids presented at a younger age and had a younger age of symptom onset, longer symptom duration, more hospitalizations, a higher frequency of diabetes mellitus, a higher pain score, and more restriction in daily activity (all P<.05). Twenty-one (46%) patients with opioid use had a total pancreatectomy compared with 9 (14%) without opioid use (P=.0002); the 21 patients also had a higher frequency of postoperative bleeding and early reoperation (8 vs 2, P<.02; 11 vs 3, P=.003, respectively). Mortality and overall morbidity was not significantly different between the 2 groups (4 vs 1, 27 vs 34, respectively). Pain scores improved postoperatively in both groups (P=.001) and was not significantly different between the groups from 12 months onward (median follow-up of 12 months, range, 3 60 months). Twenty percent of patients who used preoperative opioids however reverted to morphine use compared with 6% of patients who had not used opioids. CONCLUSIONS: Patients who used opioids had more advanced disease than patients without opioid use, accounting for part of the postoperative morbidity. Although long-term pain relief was comparable between the 2 groups, maintaining opioid withdrawal was more problematic in those with preoperative opioid use. Earlier referral for resection may be warranted in this group of patients. PMID- 15349109 TI - AMES prognostic index and extent of thyroidectomy for well-differentiated thyroid cancer in the United States. AB - BACKGROUND: The optimal extent of thyroidectomy for well-differentiated thyroid cancer is controversial. It has been suggested that surgeons use the Age, Metastases, Extent and Size (AMES) risk classification to decide between partial thyroidectomy (PT) or total thyroidectomy (TT). METHODS: The Surveillance, Epidemiology and End Results database was used to identify patients who underwent PT or TT for well-differentiated thyroid cancer between 1992 and 1997. Age, distant metastases, extrathyroidal extension, tumor size, AMES risk group, gender, histologic subtype, and lymph node metastases were analyzed by using logistic regression models to evaluate whether surgeons use these factors to determine extent of thyroidectomy. RESULTS: Of 9,226 patients, most patients had small cancers confined to the thyroid gland. Of these, 79.9% were AMES low-risk, and 83% underwent TT. Age 40 years or older (40 to 49 years: odds ratio [OR], 0.75, 95% confidence interval [CI], 0.65 to 0.86; 50 years or older: OR, 0.66; 95% CI, 0.58 to 0.75) was associated with a lower likelihood of TT, as were female gender (OR, 0.80; 95% CI, 0.70 to 0.92) and follicular histology (OR, 0.65; 95% CI, 0.55 to 0.78). Extrathyroidal extension (OR, 3.85; 95% CI, 3.09 to 4.80), regional lymph node metastases (OR, 6.98; 95% CI, 5.45 to 8.93), distant metastases (OR, 7.29; 95% CI, 2.69 to 19.8), AMES high-risk group (OR, 2.82; 95% CI, 2.36 to 3.38), and larger tumor size (OR, 1.27; 95% CI, 1.01 to 1.59) were associated with greater likelihood of TT. In multivariable analyses, only age, extrathyroidal extension, and regional and distant metastases were associated with extent of thyroidectomy; AMES risk group and tumor size were no longer significant. CONCLUSIONS: Most patients undergo TT for well-differentiated thyroid cancer regardless of AMES risk grouping. In contrast to AMES risk-group classification, younger patients are more likely to undergo TT compared with older patients. Although extrathyroidal extension and distant metastases were associated with TT in accordance with AMES criteria, tumor size had no independent influence. Regional lymph node metastasis, not an AMES criterion, increased the likelihood of TT. Some components of AMES risk-group classification are used by surgeons to choose the extent of thyroidectomy. PMID- 15349110 TI - Long-term follow-up of glucose tolerance function after pancreaticoduodenectomy: comparison between pancreaticogastrostomy and pancreaticojejunostomy. AB - BACKGROUND: The objectives of the present study are to determine the long-term changes in glucose tolerance function after pancreaticoduodenectomy and to compare the effects of pancreaticojejunostomy (PJ) and pancreaticogastrostomy (PG). Patients and methods The present study consisted of 51 patients who received a pancreaticoduodenectomy for tumors of the pancreatic head area and survived more than 7 postoperative years without tumor recurrence. According to the type of pancreatic anastomosis, they were classified into 2 groups of 25 PJ patients and 26 PG patients. Changes in the patterns of a 75-g oral glucose tolerance test (OGGT) (normal, impaired glucose tolerance [IGT], and diabetic [DM] patterns) and the need for beginning diabetic treatment (oral hypoglycemic agents or insulin) were compared between groups. RESULTS: Within 3 months after surgery, 14 (56%) patients in the PJ group had normal OGTT patterns, 8 (32%), IGT patterns, and 3 (25%), DM patterns. In the PG group, the patterns of OGTT were similar with 16 (62%) normal patterns, 6 (23%) IGT patterns, and 4 (15%) DM patterns. During the first 7 postoperative years, the 2 groups showed similar results: (1) none of the patients with normal patterns developed functional decline in glucose tolerance; (2) a high percentage of patients with initial IGT or DM patterns developed worsening glucose intolerance (7 [64%] of 11 PJ patients vs 7 [70%] of 10 PG patients); (3) the onset of functional decline in glucose tolerance occurred predominantly within the first 3 postoperative years; and (4) no specific causative event prior to the subsequent functional decline was detected. CONCLUSION: The decline of glucose tolerance after pancreaticoduodenectomy seems to be associated with a low reserve of endocrine function rather than anastomotic procedures or their related complications. Regardless of the types of pancreatic anastomosis, a close follow-up of glucose tolerance function is recommended during the first 3 postoperative years, especially among IGT or DM patients. PMID- 15349111 TI - Aprotinin inhibits local platelet trapping and improves tissue destruction in hepatic cryosurgery. AB - BACKGROUND: During the last decade, cryosurgery became an interesting alternative in the treatment of nonresectable liver neoplasms. The freeze-thaw procedure, however, may be associated with life-threatening thrombocytopenia due to local platelet trapping, and success of neoplasm ablation may be compromised by inadequate parenchymal cell destruction. METHODS: Because aprotinin is capable of inhibiting the initiation of both coagulation and fibrinolysis, we studied-by whole body scintigraphy of Indium-111-labeled platelets and histomorphology in a porcine model of hepatic cryosurgery-whether this serine protease inhibitor is effective in attenuating platelet trapping and in improving tissue destruction. RESULTS: Fifteen minutes of cryotherapy (-168 degrees C at the tip of the cryoprobe) induced a 30 +/- 4 cm(3) cryolesion, which presented with massive platelet trapping (14.0 +/- 1.7% cryolesion activity/whole body activity) and incomplete parenchymal cell destruction (0.9 +/- 0.3; score of hepatocyte nuclear destruction within the margin of the cryolesion). Aprotinin treatment with 500,000 IU initial bolus injection and additional 500,000 IU infusion over 3 hours did not affect the size of the cryolesion (29 +/- 3 cm(3)) but reduced local platelet activity (1.9 +/- 1.9%; P<.001) and induced hepatocyte nuclear destruction (3.0 +/- 0.0; P<.001). CONCLUSIONS: Thus, our study indicates that aprotinin inhibits cryoablation-associated platelet trapping and improves tissue destruction. The serine protease inhibitor may represent a valuable adjunct in cryosurgery of hepatic neoplasms. PMID- 15349112 TI - Cardiac mucosa in the remnant esophagus after esophagectomy is an acquired epithelium with Barrett's-like features. AB - BACKGROUND: The cervical esophagus is normally lined by squamous epithelium and is usually not exposed to gastroesophageal reflux. The aims of this study were, first, to investigate whether cardiac mucosa can be acquired in the remnant cervical esophagus after esophagectomy and cervical esophagogastrostomy and, second, to characterize this mucosa if present. METHODS: The medical records of 100 patients who had undergone esophagectomy with gastric pull-up reconstruction were studied retrospectively to identify those who had biopsies from the cervical esophagus proximal to the gastroesophageal anastomosis during postoperative follow-up. The histopathology and immunohistochemical stains were reviewed to assess similarity to Barrett's mucosa (cytokeratins [CK] 7 and 20 and DAS-1), cellular proliferation (topoisomerase 2alpha), and the potential for dysplasia (cyclo-oxygenase 2 [COX-2] and ornithine decarboxylase [ODC]). RESULTS: Supra anastomotic biopsies were performed in 20 patients. Cardiac mucosa was present in 10 of 20 (50%) patients in whom biopsies were performed. Four patients had areas of intestinal metaplasia, and dysplasia, and adenocarcinoma developed in 1 patient. The CK7/20 and DAS-1 staining of the columnar mucosa showed a pattern similar to Barrett's mucosa. Topoisomerase 2alpha protein expression was present in 50% of patients with cardiac mucosa. DAS-1 protein was expressed in cervical columnar mucosa but not in normal squamous esophagus mucosa. The cardiac mucosa stained weakly for COX-2 and ODC. CONCLUSIONS: Cardiac mucosa can be acquired. Its expression profile is similar to cardiac mucosa and intestinal metaplasia found in Barrett's esophagus, and different from normal esophageal or gastric mucosa. The development of cardiac mucosa is likely to be related to reflux of acid into the remnant cervical esophagus as the first step in the development of Barrett's esophagus. These findings are applicable to the development of similar changes at the gastroesophageal junction. PMID- 15349113 TI - Teaching practicing surgeons critical appraisal skills with an Internet-based journal club: A randomized, controlled trial. AB - BACKGROUND: The effectiveness of interventions for developing critical appraisal skills in practicing physicians has not been studied, despite the documented importance of reading the literature in caring for patients and in continuing professional development. The objective of this study was to evaluate whether an Internet-based intervention would lead to enhanced critical appraisal skills in practicing surgeons. METHODS: General surgeons who agreed to participate were randomized into 2 groups. The intervention was a curriculum in critical appraisal skills that included a clinical and methodologic article, a listserve discussion, and clinical and methodologic critiques. The control group received only the clinical articles. The primary outcome measure was a previously validated 2-hour test of critical appraisal. RESULTS: Of the 55 surgeons who completed the examination, subjects in the intervention group performed better on the test of critical appraisal skills than those in the control group (mean score: intervention group, 58% +/- 8 vs control group, 50% +/- 8), with a large effect size of 1.06 standard deviation units (t+3.92, P <.0001). Training conditions accounted for 22% of the variance in total scores. CONCLUSIONS: A multifaceted, Internet-based intervention resulted in improved critical appraisal skills of practicing general surgeons. PMID- 15349114 TI - Invited commentary: Assessing the evidence--filling the glass. PMID- 15349115 TI - Outcome after simultaneous colorectal and hepatic resection for colorectal cancer with synchronous metastases. AB - BACKGROUND: Consensus has not been reached concerning the timing of hepatectomy in patients with synchronous colorectal liver metastases, specifically with respect to patient selection criteria for simultaneous resection of the colorectal primary and the liver metastasis. METHODS: Retrospectively obtained clinicopathologic data for 39 consecutive patients with synchronous colorectal cancer metastases to the liver, who underwent curative simultaneous "1-stage" hepatectomy and resection of the colorectal primary at 1 institution, were subjected to univariate and multivariate analysis concerning the safety and success of the combined procedure. RESULTS: Only the volume of the resected liver was selected as a risk factor for postoperative complications (350 g mean resected liver volume in patients with postoperative complications vs 150 g in those without complications; P <.05). Patient age of 70 years or older (P <.05) and poorly differentiated or mucinous adenocarcinoma as the primary lesion (P <.01) predicted decreased overall survival by univariate analysis. Multivariate analysis retained histologic differentiation of the colorectal primary as an independent survival predictor (P <.05). CONCLUSIONS: A 1-stage procedure appears desirable for synchronous colorectal hepatic metastases except for patients requiring resection of more than 1 hepatic section, patients aged 70 years or older, and those with poorly differentiated or mucinous adenocarcinomas as primary lesions. PMID- 15349116 TI - Appraisal of concomitant splenectomy in liver resection for hepatocellular carcinoma in cirrhotic patients with hypersplenic thrombocytopenia. AB - BACKGROUND: Liver resection usually is not recommended for hepatocellular carcinoma (HCC) in cirrhotic patients with portal hypertension. The role of concomitant splenectomy in liver resection for HCC in cirrhotic patients with hypersplenic thrombocytopenia (HT) resulting from portal hypertension remains undefined. METHODS: Among 526 cirrhotic patients who underwent liver resection for HCC, 41 underwent a concomitant splenectomy (Sp group) because of HT (platelet count Hc the ferromagnetic Fe(III)2Cu(II) chains exhibit frequency dependence of the out-of-phase ac susceptibility signal at T < 4.0 K. The magnetic behavior of 4 corresponds to that of a ferromagnetically coupled chain of low spin iron(III) and copper(II) ions with frequency dependence of the out-of-phase susceptibility at T < 3.0 K. Theoretical calculations using methods based on density functional theory (DFT) have been employed to analyze and substantiate the exchange pathways in this family of complexes. PMID- 15349156 TI - A new zincoborophosphate templated by diethylenetriamine (DETA): synthesis and characterizations of (C4N3H16)-[Zn3B3P6O24].H2O. AB - The title zincoborophosphate was synthesized hydrothermally from a mixture of ZnO, B2O3, H3PO4 and the structure-directing amine, DETA. Its crystal structure contains helices made up of corner-sharing BO4 and PO4 tetrahedra, which interconnected with ZnO4 tetrahedra to form a three dimensional structure with the CZP topology. The amine and water molecules occupy the free thread of the helices and the center of the channels, respectively. Thermal stability investigations show that this new compound can be dehydrated at 423 K and the product is stable up to 723 K. Furthermore, the as-synthesized title compound and its dehydration product are luminescent, each with a similar broad band between 350 and 550 nm, but the emission of the latter is about three times more intense than that of the former. PMID- 15349157 TI - Three-component systems for conventional and window-shaped response fluorescent pH indicators. AB - Three-component systems made of a tetradentate bis-amino bis-quinoline ligand, a transition metal cation (Ni2+ or Cu2+) and a fluorescent indicator (Coumarin 343) have been studied in a water-dioxane (1 : 4 v/v) mixture, through potentiometric, pH-spectrophotometric and pH-fluorimetric titrations. For the Cu2+ containing systems, an "on-off-on" variation of fluorescence intensity vs. pH has been observed, whereas in the presence of Ni2+ a simple "on-off" profile of the fluorescence intensity vs. pH was obtained. These ternary systems thus behave as window-shaped or conventional pH-indicators, depending whether Cu2+ or Ni2+ is used as the cation. PMID- 15349158 TI - N-Salicylideneamino acidato complexes of oxovanadium(IV). The cysteine and penicillamine complexes. AB - Oxovanadium(IV) complexes with ligands derived from the reaction of salicylaldehyde with L-cysteine and with D- and D,L-penicillamine are prepared. The compounds are characterised by elemental analysis, spectroscopy (UV-VIS, CD, EPR), TG, DSC and magnetic susceptibility measurements (9-295 K). We discuss several aspects related to the structure of these complexes in the solid state and in solution; in particular, the possibility of forming thiazolidine complexes, and their comparison with the characterised complexes is studied by molecular mechanics and density functional theory calculations. The solution structures depend on pH and solvent, and while with L-Cys the spectroscopic results show trends similar to those of the L-Ala and L-Ser systems up to ca. pH 8-9, where thiolate coordination starts being detected, the penicillamine system is quite distinct, namely thiolate coordination occurs for pH > 6.5. In the presence of salicylaldehyde and V(IV)O the desulfydration of cysteine proceeds rapidly, but no similar reaction occurs with penicillamine, although its decomposition is also activated. The DFT calculations do not indicate any energetic basis for this distinct reactivity, which possibly results from different complexes present in the Cys and Pen systems. In the cysteine system, the N-salicylidenedehydroalanine-V(IV)O complex V is believed to form in an intermediate stage of the desulfydration. Further, addition of several nucleophiles to the cysteine reaction mixtures produce amino acid derivatives by a Michael-type base-catalysed addition, a result compatible with the formation of V. The products of these reactions were analysed by TLC and HPLC, and in some cases isolated. PMID- 15349159 TI - Complexation of thorium(IV) with acetate at variable temperatures. AB - The complexation between Th(IV) and acetate in 1.05 mol kg(-1) NaClO4 was studied at variable temperatures (10, 25, 40, 55 and 70 degrees C). The formation constants of five successive complexes, Th(Ac)j(4-j)+ where Ac = CH3COO- and j = 1-5, and the molar enthalpies of complexation were determined by potentiometry and calorimetry. Extended X-ray absorption fine structure spectroscopy (EXAFS) provided additional information on the complexes in solution. The effect of temperature on the stability of the complexes is discussed in terms of the electrostatic model. PMID- 15349160 TI - Speciation in the aqueous H+/H2VO4-/H2O2/citrate system of biomedical interest. AB - The speciation in the quaternary aqueous H+/H2VO4-/H2O2/citrate (Cit3-) and H+/H2VO4-/Cit3-/L-(+)-lactate (Lac-) systems has been determined at 25 degrees C in the physiological medium of 0.150 M Na(Cl). A combination of 51V NMR integral intensities and chemical shift (Bruker AMX500) as well as potentiometric data (glass electrode) have been collected and evaluated with the computer program LAKE, which is able to treat multimethod data simultaneously. The pKa-values for citric acid have been determined as 2.94, 4.34 and 5.61. Altogether six vanadate citrate species have been found in the ternary H+/H2VO4-/Cit3- system in the pH region 2-10, only two of which are mononuclear. Reduction of vanadium(V) becomes more pronounced at pH < 2. Solutions, in which reduction occurred to any extent, were excluded from all calculations. In the quaternary H+/H2VO4-/H2O2/Cit3- system, eight complexes have been found in addition to all binary and ternary complexes over the pH region 2-10, including three mononuclear species. Equilibria in general are fast, but the significant and rapid decomposition of peroxide in acidic solutions limited the final model to pH > 4. In the quaternary H+/H2VO4-/Cit3-/Lac- system, two mixed-ligand species have been determined, with the compositions V2CitLac2- and V2CitLac3- (pKa = 5.0). To our knowledge, this is the first time such complexes have been reported for vanadium(V). 51V NMR chemical shifts, compositions and formation constants are given, and equilibrium conditions are illustrated in distribution diagrams as well as the fit of the model to the experimental data. When suitable, structural proposals are given, based on 13C NMR measurements and available literature data of related compounds. PMID- 15349161 TI - Photodissociation of CO from [Ru3(mu3-O)(mu-OOCCH3)6(CO)L2] in acetonitrile, where L = pyridine, 4-cyanopyridine and methanol. AB - Photodissociation of CO from oxo-centered trinuclear ruthenium clusters [Ru3(mu3 O)(mu-OOCCH3)6(CO)L2] (L = pyridine (py): 1; 4-cyanopyridine (cpy): 2; methanol: 3) dissolved in organic solvents has been examined. Upon photolysis (> or = 290 nm, a 450-W Xe lamp), an absorption peak at 585 nm observed for 1 in CH3CN decreases its intensity and a new absorption band appears and grows at 896 nm. This spectral change, presenting isosbestic points, corresponds to photosubstitution of CO in 1 to form [Ru3(mu3-O)(mu-OOCCH3)6(CH3CN)(py)2] 4. Photoexcitation of carbonyl complexes 2 and 3 in CH3CN also affords the corresponding CH3CN-coordinated complexes [Ru3(mu3-O)(mu-OOCCH3)6(CH3CN)(cpy)2] 6 and [Ru3(mu3-O)(mu-OOCCH3)6(CH3CN)3] 7, respectively. The photosubstitution reactions (excitation wavelength, > or = 290 nm) are well described by the first order kinetics: k = 7.3 x 10(-4) s(-1) for 1, 4.9 x 10(-4) s(-1) for 2 and 5.1 x 10(-4) s(-1) for 3 (298 K). In the presence of a 100-fold excess of py, photolysis of 1 yields a tris(py) complex [Ru3(mu3-O)(mu-OOCCH3)6(py)3] 5 via photochemical loss of CO followed by coordination of py. The overall reaction (photochemical and thermal) is also confirmed by 1H NMR spectroscopy. The dissociative character of the photosubstitution is supported by negligible effects of the concentration of the entering pyridine molecule, the nature of solvents and the type of terminal monodentate ligands (other than CO) attached to the cluster. Quantum yield measurements with varied excitation wavelengths have shown that absorption bands located in the UV region (< 400 nm) play a principal role in photosubstitution, whereas an absorption band in the visible region (centered at approximately 580 nm), ascribed to an "intracluster" charge transfer, is not at all responsible for photosubstitution. PMID- 15349162 TI - Synthesis, structure and solution NMR properties of (Ph4P)[M(SeC[O]Tol)3], M = Zn, Cd and Hg. AB - Metal selenocarboxylate salts (PPh4)[M(SeC[O]Tol)3] (M = Zn (1), Cd (2) and Hg (3); Tol = C6H4-p-CH3) have been synthesized by reacting Zn(NO3)2 .6H2O, Cd(NO3)2 .4H2O or HgCl2 with (Na+)TolC[O]Se- and PPh4Cl in the ratio of 1 : 4 : 1. The structures of these compounds were determined by single-crystal X-ray diffraction methods. The crystal structures contain discrete cations and anions. In the each anion, the metal center is bound to three TolC[O]Se ligands, primarily through Se, though some long M...O interactions also occur. NMR spectra (113Cd, 199Hg and 77Se, as appropriate) are reported for solutions of [M(SeC[O]Tol)3]-, and of [M(SeC[O]Tol)3](-) - [M(SC[O]Ph)3]- mixtures (M = Zn-Hg), in CH2Cl2 at reduced temperatures. In addition, ESI-MS data have been obtained for [M(SeC[O]Tol)(3)]( ) - [M(SC[O]Ph)3]- mixtures (M = Zn-Hg) in acetone and in CH2Cl2. The NMR and ESI MS studies show that the complexes [M(SeC[O]Tol)n(SC[O]Ph)(3-n)]- (n= 3-0) persist in solution. PMID- 15349163 TI - The reactivity of cyclo-(P5tBu4)- towards group 13, 14 and 15 metal chlorides: complexation and formation of cyclooligophosphanes, [cyclo-(P5tBu4)]2 and [cyclo (P4tBu3)PtBu]2, by reductive elimination. AB - [Na(THF)4][cyclo-(P5tBu4)] (1) reacts with Et2AlCl and GeCl4 to give Et2Al[cyclo (P5tBu4)](THF) (2) and, in low yield, GeCl3[cyclo-(P5tBu4)], respectively, while the reaction of 1 with SnCl2, PbCl2 or BiCl3 results in the formation of the structural isomers [cyclo-(P5tBu4)]2 (3) and [cyclo-(P4tBu3)PtBu]2 (4)(besides other cyclic phosphanes) and elemental metal. PMID- 15349164 TI - Extended coordination frameworks incorporating heterobimetallic squares. AB - The molecular structure of aluminium and iron(III) complexes with 3-phenyl and 3 (4-pyridyl) (HL) substituted acetylacetonate ligands is appreciably distorted. For AlL3 and FeL3 this shows that the orientation of the side pyridyl-N donor atoms lone pairs is about 90 and 135 degrees which favours the assembly of heterobimetallic square patterns in Al(Fe)L3 complexes with metal ions. This was employed for the modular construction of semi-regular heterobimetallic networks, in which the pyridyldiketonate ligands bridge pairs of Fe(Al)/Cd(Co) metal ions and support the structure of 1D and 2D coordination polymers. The unprecedented 2D structure of [Cd[AlL3](CH3OH)[NO3]2].2CHCl3 and Cd[AlL3](CH3OH)Br2].2CHCl3 . 2CH3OH is based upon plane tiling by a set of heterobimetallic squares and octagons, while [Cd[FeL3]2(NO3)2].2H2O and [Co[AlL3]2Cl2].4CHCl3 . 2CH3OH are 1D polymers and exist as chains of heterobimetallic squares sharing opposite vertices. PMID- 15349165 TI - The formation of dimeric phosph(III)azane macrocycles [[P(mu-NtBu)]2.LL]2 [LL = organic spacer]. AB - The dimeric macrocycles [[P(mu-NtBu)]2.LL]2 [LL = OCH2C(Me)2CH2O (1), 2,6 (NH)2C5H3N (2), 1,2-(NH)2C6H4(3)] have been obtained by the reactions of the appropriate diols and diamines (LLH2) with the dimeric phosph(III)azane [ClP(mu NtBu)]2. Under different conditions the reaction of 1,2-(NH2)2C6H4 with [ClP(mu NtBu)]2 gives the monomer [[P(mu-NtBu)]2.[1,2-(NH)2C6H4]] (4) (instead of the dimer 3). Contrary to the literature, the results illustrate that the formation of dimeric macrocycles is common in these reactions and dependent among other factors on the steric demands and length of the organic spacer (LL) as well as the reaction conditions. PMID- 15349166 TI - Mechanistic studies on oxidation of hydrazine by a mu-oxo diiron(III,III) complex in aqueous acidic media-proton coupled electron transfer. AB - [Fe2(mu-O)(phen)4(H2O)2]4+ (1), one of the simplest mu-oxo diiron(III) complexes, quantitatively oxidises hydrazine to dinitrogen and itself is reduced to two moles of ferroin, [Fe(phen)3]2+ in presence of excess phenanthroline. The weak dibasic acid, 1 (pKa1= 3.71 +/- 0.05 and pKa2= 5.28 +/- 0.10 at 25.0 degrees C, I= 1.0 mol dm(-3)(NaNO3)) and its conjugate bases, [Fe2(mu-O)(phen)4(H2O)(OH)]3+ (2) and [Fe2(mu-O)(phen)4(OH)2]2+ (3) are involved in the redox process with the reactivity order 1 > 2 > 3 whereas N2H4 and not N2H5+ was found to be reactive in the pH interval studied 3.45-5.60. Cyclic voltammetric studies indicate poor oxidizing capacity of the title substitution-labile diiron complex, yet it oxidizes N2H4 with a moderate rate--a proton coupled electron transfer (1e, 1H+) drags the energetically unfavourable reaction to completion. The rate retardation in D2O media is substantially higher at higher pH due to the increasing basicity of the oxo-ligand in the order 3 > 2 > 1. Marcus calculations result an unacceptably high one-electron self-exchange rate for the iron center indicating an inner-sphere nature of the electron-transfer. PMID- 15349167 TI - One- or two-dimensional 2,3-naphtho crown ether complexes [Na(N15C5)]2[M(SCN)4] and [K(N18C6)]2[M(SCN)4] (M = Pd, Pt) constructed by pi-pi stacking interactions. AB - Four novel 2,3-naphtho-15-crown-5 (N15C5) and 2,3-naphtho-18-crown-6 (N18C6) complexes [Na(N15C5)]2[Pd(SCN)4] (1), [Na(N15C5)]2[Pt(SCN)4] (2), [K(N18C6)]2[Pd(SCN)4] (3) and [K(N18C6)]2[Pt(SCN)4] (4) were synthesized and characterized by elemental analysis, FT-IR spectra and single-crystal X-ray diffraction. The structure analyses reveal that both 1 and 2 are assembled into zigzag chains by the strong intermolecular pi-pi stacking interactions between adjacent 2,3-naphthylene groups of N15C5. The molecules of complexes 3 and 4 are linked into 1D chains by the bridging K-O(ether) interactions between the adjacent [K(N18C6)]+ units and the resulting chains are constructed into a novel 2D network by inter-chain pi-pi stacking interactions between the neighboring 2,3 naphthylene moieties of N18C6. According to the supramolecular self-assemblies of complexes 1-4, two types of stacking model of naphthylene groups are given and discussed. PMID- 15349168 TI - Synthesis, structure and luminescent properties of yttrium benzene dicarboxylates with one- and three-dimensional structure. AB - The synthesis and structure of two yttrium benzene dicarboxylates, 1 is proportional to [[Y2(C12N2H8)2(C8H4O4)3].H2O], I and 3 is proportional to [[Y2(C12N2H8)2(C8H4O4)3]], II with one- and three-dimensional structure has been accomplished employing hydrothermal methods in the presence of 1,10 phenanthroline. While I is formed with phthalic aid (1,2-BDC), II is formed using isophthalic acid (1,3-BDC). Both the structures appear to have comparable building units, an eight-membered ring and a paddle-wheel arrangement, connected through the carboxylic acid. The 1,10-phenanthroline, connected to Y as a secondary ligand, occupies the inter-chain spaces in I, and projects into the channels in II. The channels in II are inter-connected. Photoluminescence studies indicate that both I and II exhibit a bathochromic shift with respect to the acids (1,2-BDC and 1,3-BDC) and a hypsochromic shift with respect to 1,10 phenanthroline. Both the compounds exhibit reasonable pi...pi interactions. PMID- 15349169 TI - Dinuclear 1,4,7-triazacyclononane (tacn) complexes of cobalt(III) with amido and tacn bridges. Synthesis, characterization and reversible acid-accelerated bridge cleavage. AB - Amido-bridged dinuclear cobalt(III) complexes with 1,4,7-triazacyclononane (tacn) were synthesized from [Co(tacn)(O3SCF3)3] by treatment with potassium amide in liquid ammonia at 100 degrees C. Two isomeric triply bridged complexes, [(tacn)Co(mu-NH2)3Co(tacn)]3+ and [(tacn)Co(mu-NH2)2[mu-tacn(-H)]Co(NH3)]3+, were isolated as perchlorates, and the crystal structure of the perrhenate of the latter complex was determined by X-ray diffraction. In this compound a nitrogen atom (deprotonated) from one of the tacn ligands forms a third bridge together with two amido bridges. In 1.0 M (Na,H)ClO4 ([H+] 0.1-1.0 M) the two isomers undergo acid-accelerated amido bridge cleavage, as earlier found for chromium(III) analogues, in spite of the fact that such bridges are co ordinatively saturated. The triamido-bridged isomer is in this acid medium in equilibrium with [(H2O)(tacn)Co(mu-NH2)2Co(tacn)(NH3)]4+. An isolated perchlorate of this complex appeared to be the salt of the trans-ammineaqua isomer as determined by X-ray diffraction. Equilibration from both sides fits the first order rate constant dependence k(obs)=6.2(3) x 10(-5)[H+] + 2.1(2) x 10(-5)(s( 1)) at 40 degrees C. Prolonged treatment of the two triply bridged isomers in 1.0 M HClO4 at elevated temperature produces primarily triply bridged dinuclear species where one or two amido bridges have been replaced by hydroxo bridges. PMID- 15349170 TI - Syntheses, spectroscopic and molecular quadratic nonlinear optical properties of dipolar ruthenium(II) complexes of the ligand 1,2-phenylenebis(dimethylarsine). AB - Six new complex salts trans-[Ru(II)Cl(pdma)2L][PF6]n [pdma = 1,2 phenylenebis(dimethylarsine); L = (E,E,E)-1,6-bis(4-pyridyl)hexa-1,3,5-triene (bph), n= 1, 5; L =N-methyl-4-[(E)-2-(4-pyridyl)ethenyl]pyridinium (Mebpe+), n= 2, 7; L =N-methyl-4-[(E,E)-4-(4-pyridyl)buta-1,3-dienyl]pyridinium (Mebpb+), n= 2, 8; L =N-methyl-4-[(E,E,E)-6-(4-pyridyl)hexa-1,3,5-trienyl]pyridinium (Mebph+), n= 2, 9; L = bis(4-pyridyl)acetylene (bpa), n= 1, 10; L =N-methyl-4-[2-(4 pyridyl)ethynyl]pyridinium (Mebpa+), n= 2, 11] have been prepared. The electronic absorption spectra of 5 and 7-11 display intense, visible metal-to-ligand charge transfer (MLCT) bands, with lambdamax values in the range 434-492 nm in acetonitrile. Cyclic voltammetric studies reveal reversible Ru(III/II) waves with E(1/2) values in the range 1.06-1.15 V vs. Ag-AgCl, together with irreversible L based reduction processes. Along with a number of previously reported related compounds (B. J. Coe et al., J. Chem. Soc., Dalton Trans., 1996, 3917; 1997, 591; 2000, 797), salts 5 and 7-11 have been investigated by using Stark (electroabsorption) spectroscopy in butyronitrile glasses at 77 K. These studies have afforded dipole moment changes Deltamu12 for the MLCT transitions which have been used to calculate molecular static first hyperpolarizabilities beta0 according to the two-state equation beta0= 3Deltamu12(mu12)2/(Emax)2 (mu12 = transition dipole moment, Emax = MLCT energy). MLCT absorption and electrochemical data show that a trans-[Ru(II)Cl(pdma)2]+ centre is considerably less electron-rich than a [Ru(II)(NH3)5]2+ unit. Although the beta0 responses of the pdma complexes are only a little smaller than those of their [Ru(II)(NH3)5]2+ analogues, this result is partly attributable to unexpected changes in the relative mu12 values on freezing. Thus, substantial increases in mu12 for the arsine compounds act to partially offset the beta0-decreasing influence of their higher Emax values when compared with the analogous pentaammine species. Single crystal X-ray structures have been obtained for the salts 1(.)2.5MeCN, 4(.)2MeCN, 7 and 11, but only 1(.)2.5MeCN adopts a non-centrosymmetric space group (Fdd2) such as may show bulk NLO effects. PMID- 15349171 TI - Alkyl-eta2-alkene niobocene and tantalocene complexes with the allyldimethylsilyl eta5-cyclopentadienyl ligand: synthesis, NMR studies and DFT calculations. AB - Group 5 metal complexes [M(eta5-C5H5)[eta5-C5H4SiMe2(CH2-eta]2-CH=CH2)]X] (M = Nb, X = Me, CH2Ph, CH2SiMe3; M = Ta, X = Me, CH2Ph) and [Ta(eta5-C5Me5)[eta5 C5H4SiMe2(CH2-eta2-CH=CH2)]X] (X = Cl, Me, CH2Ph, CH2SiMe3) containing a chelating alkene ligand tethered to a cyclopentadienyl ring have been synthesized in high yields by reduction with Na/Hg (X = Cl) and alkylation with reductive elimination (X = alkyl) of the corresponding metal(iv) dichlorides [M(eta5 Cp)[eta5-C5H4SiMe2(CH2CH=CH2)]Cl2] (Cp = C5H5, M = Nb, Ta, Cp = C5Me5, M = Ta). These chloro- and alkyl-alkene coordinated complexes react with CO and isocyanides [CNtBu, CN(2,6-Me2C6H3)] to give the ligand-substituted metal(III) compounds [M(eta5-Cp)[eta5-C5H4SiMe2(CH2CH=CH2)]XL] (X = Cl, Me, CH2Ph, CH2SiMe3). Reaction of the chloro-alkene tantalum complex with LiNHtBu results in formation of the imido hydride derivative [Ta(eta5-C5Me5)[eta5 C5H4SiMe2(CH2CH=CH2)]H(NtBu)]. NMR studies for all of the new compounds and DFT calculations for the alkene-coordinated metal complexes are compared with those known for related group 4 metal cations. PMID- 15349172 TI - Synthesis and reactivity of iridium complexes with pyridine and piperidine ligands: models for hydrodenitrogenation. AB - The complexes [Ir(H)2(eta1-N-L)2(PPh3)2]PF6, L = py (1), iQ (2) and pip (3) (py = pyridine, iQ = isoquinoline, pip = piperidine) have been synthesized in high yields by hydrogenation of [Ir(cod)(PPh3)2]PF6 in the presence of the appropriate nitrogen compound. When hydrogen is bubbled through 1,2-dichloroethane solutions of 1 or 2, two new species were formed in each case by C-Cl bond activation of the solvent, Ir(H)2Cl(eta1-N-L)(PPh3)2 (L = py, 4; iQ, 5) and IrH(Cl)2(eta1-N L)(PPh3)2 (L = py, 6; iQ, 7). Reaction of 3 with py or iQ yielded complexes 1 and 2, respectively, while under a slow stream of carbon monoxide the complex [Ir(H)2(eta1-N-pip)(CO)(PPh3)2]PF6 (8) was produced. Complex 3 also reacts with halide and 4-bromothiophenolate anions leading to the corresponding neutral species Ir(H)2(X)(eta1-N-pip)(PPh3)2, X = Cl (9), I (10) and 4-BrC6H4S (11), or with [MoS4]2- to yield the hetero-bimetallic complex [Ir(H)(PPh3)2(mu-S)2MoS2]- (13). All the new complexes were characterized by analytical and spectroscopic methods. The X-ray structures of , 2 and 8 consist of distorted octahedra with a mutually cis disposition of the two hydrides and mutually trans phosphines. Complexes 1, 2 and 3 and their derivatives are of interest as models for the chemisorption step in hydrodenitrogenation reactions on solid catalysts. PMID- 15349173 TI - EPR study of intramolecular dynamics in o-semiquinonic nickel complexes with PCP pincer ligand in solution. AB - A number of square pyramidal o-semiquinonic nickel complexes with 2,6 bis(diphenylphosphinomethyl)phenyl were investigated by EPR spectroscopy in solution. The complexes have flexible coordination spheres and can exist in solution as two isomers rapidly (on the EPR timescale) interconverting one to another at room temperature. The equilibrium is entropy driven and does not significantly depend on the substituents in the semiquinone ring. The spin density distribution depends on mesomeric properties and bulkiness of substituents in ortho-position to oxygen atoms. PMID- 15349174 TI - [Rh(I)L]-catalyzed cyclotetramerization of 1,3-butadiene: a theoretical investigation of alternative mechanistic paths for the generation of the [Rh(III)(octadienediyl)(PR3)]+ complex. AB - A detailed theoretical investigation of alternative mechanistic paths for the formation of the [Rh(III)(octadienediyl)(PiPr3)]+ complex is presented, employing a gradient-corrected density functional theory (DFT) method (BP86). This process represents most likely the first step in the recently reported [Rh(I)L]-catalyzed cyclotetramerization of butadiene (M. Bosch, M. S. Brookhart, K. Ilg and H. Werner, Angew. Chem., Int. Ed., 2000, 39, 2304). The favorable route for oxidative addition under C-C-bond formation starts from the prevalent [Rh(I)(butadiene)2(PiPr3)]+ form of the active catalyst through oxidative coupling between two cis-eta4-butadienes. This affords the [Rh(III)(bis-eta3-anti octadienediyl)(PiPr3)]+ compound as the kinetic coupling product that consecutively undergoes transformation into the thermodynamically favorable bis eta3-syn-octadienediyl-Rh(III) isomer via facile allylic conversions occurring in the octadienediyl framework. The computationally predicted energy profile is almost in quantitative agreement with the experimentally determined kinetics and allows a consistent rationalization of the experimental observations. PMID- 15349175 TI - Quasi-octahedral complexes of pentamethylcyclopentadienyliridium(III) bearing bis(diphenylphosphinomethyl)phenylphosphine (dpmp). AB - Reaction of [Cp*IrCl2]2 (1) with dpmp in the presence of KPF6 afforded a binuclear complex [Cp*IrCl(dpmp-P1,P2;P3)IrCl2Cp*](PF6) (2) (dpmp =(Ph2PCH2)2PPh). The mononuclear complex [Cp*IrCl(dpmp-P1,P2)](PF6) (4) was generated by the reaction of [Cp*IrCl2(BDMPP)](BDMPP =PPh[2,6-(MeO)2C6H3]2) with dpmp in the presence of KPF6. These mono- and binuclear complexes have four membered ring structures with a terminal and a central P atom of the dpmp ligand coordinated to an iridium atom as a bidentate ligand. Since there are two chiral centers at the Ir atom and a central P2 atom, there are two diastereomers that were characterized by spectrometry. Complexes anti-4 and syn-4 reacted with [Cp*RhCl2]2 or [(C6Me6)RuCl2]2, giving the corresponding mixed-metal complexes, anti- and syn- [Cp*IrCl(dppm-P1,P2;P3)MCl2L](PF6) (6: M = Rh, L = Cp*; 7: M = Ru, L = C6Me6). Treatment with AuCl(SC4H8) gave tetranuclear complexes, anti- and syn 8 [[Cp*IrCl(dppm-P1,P2;P3)AuCl]2](PF6)2 bearing an Au-Au bond. Reaction of anti- with PtCl2(cod) generated the trinuclear complex anti-9, anti-[[Cp*IrCl(dppm P1,P2;P3)]2PtCl2](PF6)2. These reactions proceeded stereospecifically. The P,O chelated complex syn-[Cp*IrCl(BDMPP-P,O)] (syn-10)(BDMPP-P,O = PPh[2,6 (MeO)2C6H3][2-O-6-(MeO)C6H3]2) reacted with dpmp in the presence of KPF6, generating the corresponding anti-complex as a main product as well as a small amount of syn-complex, [Cp*Ir(BDMPP-P,O)(dppm-P1)](PF6) (11). The reaction proceeded preferentially with inversion. The reaction processes were investigated by PM3 calculation. anti- was treated with MCl2(cod), giving anti-[Cp*Ir(BDMPP P,O)(dppm-P1;P2,P3)MCl2](PF6)(14: M = Pt; 15: M = Pd), in which the MCl2 moiety coordinated to the two free P atoms of anti-11. The X-ray analyses of syn-2, anti 2, anti-4, anti-8 and anti-11 were performed. PMID- 15349502 TI - Receptor tyrosine kinases and inhibitors in lung cancer. AB - Lung cancer is a deadly disease with high mortality and morbidity. Most cases of lung cancer are due to non-small cell carcinoma, with 16% of cases being small cell carcinoma. The biology at a cellular level is of interest at many levels. Knowing cellular pathways helps to further enhance our knowledge of how lung cancer cells survive, proliferate, and metastasize. The receptor tyrosine kinases (RTKs) located at the cellular membrane are becoming of great interest as sites for targeted therapies for lung cancers. This review will discuss the RTKs that are involved in lung cancers and the newer therapies that are being tested. We will specifically discuss receptors such as epidermal growth factor receptor, c Kit receptor, VEGF receptor, c-Met receptor, insulin growth factor receptor, and Eph receptor. The inhibitors against the specific RTKs are in various preclinical and clinical trials, and this will be detailed. PMID- 15349503 TI - Collagenous tissues upon lithium treatment: a quantitative ultrastructural study. AB - In this review, the influence of lithium treatment in mouse, rat, and rabbit skin, liver, bone, and aorta, as well as arachnoid and dura mater collagen fibrils, is examined using electron microscopy and image processing. Structural changes (fibril architecture and diameter) are detected at the ultrastructural level in specimens from all lithium-treated tissues. The overall collagen fibril architecture is disturbed as compared with specimens from normal species. The mean diameter values of treated collagen fibrils are significantly smaller than those from controls in all tissues examined. The banding patterns of fibrils are normal in all cases. Measurements by a computerized method of measuring axial periodicity of fibrils indicate no effect of lithium on this parameter. Computer analysis shows no differences in charged amino acid composition between lithium treated and -untreated samples. Under the present experimental conditions, lithium can induce permanent structural collagen alterations. PMID- 15349504 TI - Detection and characterization of beta-lactam resistance in Bacillus cereus PTCC 1015. AB - In the present study, detection, isolation, and characterization of beta lactamases from Bacillus cereus PTCC 1015 were investigated. B. cereus was inoculated in nutrient broth containing ampicillin (50 mg x ml(-1)) for 24 h (35 degrees C, 200 rpm). Activity measurements were carried out against ampicillin (0.1 mg x ml(-1)) and cephalexin (0.08 mg x ml(-1)) by a spectrophotometric method at different conditions (pH 6-10, temperatures 25-45 degrees C). Maximum penicillinase and cephalosporinase activity was observed at pH 7. The optimized temperatures for penicillinase and cephalosporinase activity were 30 and 40 degrees C, respectively. At the above conditions, maximum enzymatic activity was calculated as 0.89 +/- 0.014 and 0.037 +/- 0.001 units against ampicillin and cephalexin. PMID- 15349505 TI - RNA editing in hepatitis delta virus: unsolved puzzles. AB - RNA editing, or post-transcriptional changes in the sequences of RNAs, is being increasingly recognized as an important player in the regulation of gene expression in vertebrates and invertebrates. Different types of RNA editing have been reported. This review discuss the type of RNA editing caused by cellular enzymes known as adenosine deaminases that act on RNAs (ADARs), and it"s significance in the lifecycle of an RNA virus, hepatitis delta virus. PMID- 15349506 TI - Clinical holistic medicine: a psychological theory of dependency to improve quality of life. AB - In this paper, we suggest a psychological theory of dependency as an escape from feeling existential suffering and a poor quality of life. The ways in which human beings escape hidden existential pains are multiple. The wide range of dependency states seems to be the most common escape strategy used. If the patient can be guided into the hidden existential pain to feel, understand, and integrate it, we believe that dependency can be cured. The problem is that the patient must be highly motivated, sufficiently resourceful, and supported to want such a treatment that is inherently painful. Often, the family and surrounding world is suffering more than the dependent person himself, because the pattern of behavior the patient is dependent on makes him or her rather insensitive and unable to feel. If the patient is motivated, resourceful, and trusts his physician, recovery from even a severe state of dependency is not out of reach, if the holistic medical tools are applied wisely. The patient must find hidden resources to take action, then in therapy confront and feel old emotional pain, understand the source and inner logic of it, and finally learn to let go of negative attitudes and beliefs. In this way, the person can be healed and released of the emotional suffering and no longer be a slave to the dependency pattern. PMID- 15349507 TI - Attitudes of Bedouin and Jewish physicians towards the medical care for persons with intellectual disability in the Bedouin Negev community. A pilot study. AB - Change in the attitudes of staff or the public towards people with intellectual disability (ID) can impact their life and health, but that change has not been studied among physicians who belong to an ethnic minority undergoing dramatic social and economic transition. The goal of this study was to explore the change of attitudes of Negev Bedouin physicians serving their community and their satisfaction with policy, care, and knowledge in the field of ID. Seventeen community physicians (7 Bedouins and 10 Jewish) were interviewed using a simple questionnaire that consisted of items measuring attitude and satisfaction. The vast majority of the Bedouin and Jewish physicians had positive attitudes toward inclusion of those in the community with ID and were ready to provide the care needed in the community with special assistance. There was a need for further education in ID and more resources. There was a belief that there is discrimination between the Bedouin and Jewish community in the provision of care to people with ID. General dissatisfaction was expressed about the policy, resources, care provision, and expertise offered to Bedouins with ID. More efforts must be directed to empower the physicians with knowledge, expertise, and resources to handle the care of Bedouins with ID in a culturally appropriate way. PMID- 15349508 TI - Buried rubber band: a cause of persistent penile discharge, recurrent urinary tract infections, and development of urethrocutaneous fistula. PMID- 15349509 TI - Bio-liquid morphological analysis. AB - Information is presented on the new scientific line in medicine and biology: bio liquid morphology. The interdisciplinary character of the given research area is emphasized. The problems and prospects of bio-liquid morphological analysis development both in applied and fundamental aspects are discussed. PMID- 15349510 TI - Embryos, clones, and stem cells: a scientific primer. AB - This article is intended to give the nonspecialist an insight into the nuances of "clones", cloning, and stem cells. It distinguishes embryonic and adult stem cells, their normal function in the organism, their origin, and how they are recovered to produce stem cell lines in culture. As background, the fundamental processes of embryo development are reviewed and defined, since the manipulation of stem cell lines into desired specialized cells employs many of the same events. Stem cells are defined and characterized and shown how they function in the intact organism during early development and later during cell regeneration in the adult. The complexity of stem cell recovery and their manipulation into specific cells and tissue is illustrated by reviewing current experimentation on both embryonic and adult stem cells in animals and limited research on human stem cell lines. The current and projected use of stem cells for human diseases and repair, along with the expanding methodology for the recovery of human embryonic stem cells, is described. An assessment on the use of human embryonic stem cells is considered from ethical, legal, religious, and political viewpoints. PMID- 15349511 TI - Clinical holistic medicine: treatment of physical health problems without a known cause, exemplified by hypertension and tinnitus. AB - In the medical clinic, we often face health problems that have no known cause, even after a thorough examination. Biomedicine is often unable to find a cure in these situations, leaving the problem unsolved or leaving the patient on a palliative pharmaceutical cure, which is often for a lifetime. In this case, consciousness-based, holistic medicine could be an alternative. Using the theories and tools of holistic medicine wisely, the physician can often provide treatment for the patient. The toolbox of holistic medicine makes it possible to work on everybody because there is always something related to the patients quality of life that can be improved: his love, his purpose of life, and the way he uses his talents, his mind, his feelings, his body, and his sexuality. For treatment in holistic medicine, it really does not matter as much that you cannot give the patient a precise medical diagnosis, because you can always work on the patient with the intention of healing his or her whole life and existence. It is quite a paradox that many of these diseases can be understood on the level of the individual patient at the same moment that the patient is cured; many of these diseases seem to be clearly related to the repression of the individual character, as stressed already by Hippocrates. So if you simply start working with the patient to help him confront old existential pain and coach him in his personal development of his life by intensifying its meaning and purpose, the symptoms very often simply disappear. The toolbox of holistic medicine also seems relevant to even difficult, neurotic, psychosomatic, and hypochondriac patients. Believing in the treatment and not giving up on your patient, and moving forward in the treatment with the patient himself is the ultimate goal, even when you yourself do not understand the mechanism fully. This will force you to develop your own competence and is, in essence, what makes an outstanding holistic physician. PMID- 15349512 TI - A photoluminescence-based field method for detection of traces of explosives. AB - We report a photoluminescence-based field method for detecting traces of explosives. In its standard version, the method utilizes a commercially available color spot test kit for treating explosive traces on filter paper after swabbing. The colored products are fluorescent under illumination with a laser that operates on three C-size flashlight batteries and delivers light at 532 nm. In the fluorescence detection mode, by visual inspection, the typical sensitivity gain is a factor of 100. The method is applicable to a wide variety of explosives. In its time-resolved version, intended for in situ work, explosives are tagged with europium complexes. Instrumentation-wise, the time-resolved detection, again visual, can be accomplished in facile fashion. The europium luminescence excitation utilizes a laser operating at 355 nm. We demonstrate the feasibility of CdSe quantum dot sensitization of europium luminescence for time resolved purposes. This would allow the use of the above 532 nm laser. PMID- 15349513 TI - Metabolism of mycotoxins, intracellular functions of vitamin B12, and neurological manifestations in patients with chronic toxigenic mold exposures. A review. AB - This paper evaluates the possible reasons for consistent vitamin B12 deficiency in chronic toxigenic mold exposures and the synergistic relationships with the possible mycotoxic effects on one-carbon metabolism that lead to the manifestations of clinical neuropathological symptomology. Vitamins are first defined in general and the nutritional sources of vitamin B12 are evaluated in particular. Since patients with chronic exposures to toxigenic molds manifest vitamin B12 deficiencies, the role of mycotoxins in vitamin B12 metabolism is assessed, and since vitamin B12 plays important biochemical roles in one-carbon metabolism, the synergistic effects with mycotoxins on humans are reviewed. An outline of the proposed mechanism by which mycotoxins disrupt or interfere with the normal functions of vitamin B12 on one-carbon metabolism is proposed. The overall functions of vitamin B12 as a source of coenzymes, in intracellular recycling of methionine, in methionine synthase reaction, in the prevention of chromosome breakage, in methylation, and in maintaining a one-carbon metabolic balance are reviewed. Signs, symptoms, and clinical neurological indications of vitamin B12 deficiency are also cited. By implication and derivation, it is likely that the interruption of the structure and function of vitamin B12 would in turn interfere with the one-carbon metabolism leading to the neurological manifestations. This review is an attempt to formulate a basis for an ongoing research investigation on the subject. PMID- 15349514 TI - Using Health Utility Index (HUI) for measuring the impact on health-related quality of Life (HRQL) among individuals with chronic diseases. AB - Quality of life is an important indicator in assessing the burden of disease, especially for chronic conditions. The Health Utilities Index (HUI) is a recently developed system for measuring the overall health status and health-related quality of life (HRQL) of individuals, clinical groups, and general populations. Using the HUI (constructed based on eight attributes: vision, hearing, speech, mobility, dexterity, cognition, emotion, and pain/discomfort) to measure the HRQL for chronic disease patients and to detect possible associations between HUI system and various chronic conditions, this study provides information to improve the management of chronic diseases. This study is of interest to data analysts, policy makers, and public health practitioners involved in descriptive clinical studies, clinical trials, program evaluation, population health planning, and assessments. Based on the Canadian Community Health Survey (CCHS) for 2000-01, the HUI was used to measure the quality of life for individuals living with various chronic conditions (Alzheimer/other dementia, effects of stroke, urinary incontinence, arthritis/rheumatism, bowel disorder, cataracts, back problems, stomach/intestinal ulcers, emphysema/COPD, chronic bronchitis, epilepsy, heart disease, diabetes, migraine headaches, glaucoma, asthma, fibromyalgia, cancers, high blood pressure, multiple sclerosis, thyroid condition, and other remaining chronic diseases). Logistic Regression Model was employed to estimate the associations between the overall HUI scores and various chronic conditions. The HUI scores ranged from 0.00 (corresponding to a state close to death) to 1.00 (corresponding to perfect health); negative scores reflect health states considered worse than death. The mean HUI score by sex and age group indicated the typical quality of life for persons with various chronic conditions. Logistic Regression results showed a strong relationship between low HUI scores (< or = 0.5 and 0.06-1.0) and certain chronic conditions. Age- and sex-adjusted Odds Ratio (OR) and p values showed an effect among individuals diagnosed with each chronic disease on the overall HUI score. Results of this study showed that arthritis/rheumatism, heart disease, high blood pressure, cataracts, and diabetes had a severe impact on HRQL. Urinary incontinence, Alzheimer/other dementia, effects of stroke, cancers, thyroid condition, and back problems have a moderate impact. Food allergy, allergy other than food, asthma, migraine headaches, and other remaining chronic diseases have a relatively mild effect. It is concluded that major chronic diseases with significant health burden were associated with poor HRQL. The HUI scores facilitate the measurement and interpretation of results of health burden and the HRQL for individuals with chronic diseases and can be useful for development of strategies for the prevention and control of chronic diseases. PMID- 15349515 TI - Pseudotumoral aspect of the hydatic cyst of the kidney. PMID- 15349516 TI - Evaluation of the drug treatment and persistence of onychomycosis. AB - Onychomycosis is a common nail disease responsible for approximately 50% of diseases of the nail. It occurs more in the elderly, though several cases have been reported among children. Several factors influence, such as climate, geography, and migration. The two dermatophytes most commonly implicated in onychomycosis are Trichophyton rubrum and T. mentagrophytes, accounting for more than 90% of onychomycoses. Nonetheless, several other toxigenic molds have been implicated. For convenience, onychomycosis is divided into four major clinical presentations: distal subungal, which is the most common form of the disease; proximal subungal, which is the most common form found in patients with human immunodeficiency virus infection; superficial; and total dystrophic onychomycosis. Epidemiology of onychomycosis in adults and children is evaluated and the most common clinical symptoms addressed. Although the risk factors are discussed, the multifactorial nature of onychomycosis makes this inexhaustible. The diagnosis and treatments are difficult and the choice of appropriate antifungal drugs complex and require the knowledge of the chemical structures of the metabolites of the molds that cause onychomycosis and their interaction with the antifungal drugs. This is true because most of the antifungal drugs are derived from mold/fungal metabolism. Treatment with griseofulvin and amphotericin is displaced by the use of newer drugs from azole compounds, pyrimidines, and allylamines derivatives. Amorolfine, itraconazole, and ciclopirox nail lacquer solution 8 have gained support globally, but the side effects, drug resistance, and persistence of the disease are still a serious concern to the patients, just as economics and quality of life. Hence, the search for safer and more efficacious drug treatments are continuing. PMID- 15349517 TI - Seasonal evaluation of antigenic bacterial infections among working class in the inner city of Houston. AB - This paper evaluates the monthly, quarterly, and seasonal variation of antigenic bacterial infections among the working class in the inner city of Houston using the Wellcogen Rapid Test methods. One of the aims was to demonstrate how this method could be used effectively in screening patients at risk and preventing the spread of antigenic bacteria such as Streptococcus pneumoniae, Haemophilus influenzae b, Streptococcus (Strep b), and Neisseria meningitidis (mainly group c and b). A total of 2,837 patients were screened for bacterial infections; 908 (32%) were male and 1,929 (68%) were female. The age range was between 2 and 70 years. Of the total group, 356 (12.5%) patients were positive; 203 (57%) were female while 153 (43%) were male (male/female ratio of 1:1.3). Medically underserved and immune suppressed populations are the most affected by these bacterial infections. Blacks are the most affected (48%) compared to Native Americans (1%), but children under 10 years of age have the highest incidence. This research showed, in addition, that the Wellcogen Rapid Tests are effective (356 cases identified) for a rapid screening of infectious bacteria. Explanation for these results was probably due to poor living conditions, poor hygiene, and viral immune suppression in adults and immature immune systems in neonates and children under 10 years of age. PMID- 15349518 TI - Estimation of the cellular antioxidant response to chromium action using ESR method. AB - In the present study, the antioxidant capacity of chromium-treated L-41 (human epithelial-like cells) was investigated by the ESR spin-trapping technique. The crude cell extracts of the cells grown in the presence of 2 microM (nontoxic) and 20 microM (toxic) chromium (VI) concentrations were tested in the model Fenton system with and without catalase-inhibitor sodium azide. The presented approach using the ESR technique along with inhibitors lets us discern cell extract defense capacity connected with the enzymatic activity in viable cells and the catabolic activity in dying cells. PMID- 15349519 TI - Ammonia emission and deposition in Scotland and its potential environmental impacts. AB - The main source of atmospheric ammonia (NH3) in Scotland is livestock agriculture, which accounts for 85% of emissions. The local magnitude of emissions therefore depends on livestock density, type, and management, with major differences occurring in various parts of Scotland. Local differences in agricultural activities therefore result in a wide range of NH3 emissions, ranging from less than 0.2 kg N ha(-1) year(-1) in remote areas of the Scottish Highlands to over 100 kg N ha(-1) year-1 in areas with intensive poultry farming. Scotland can be divided loosely into upland and lowland areas, with NH3 emission being less than and more than 5 kg N ha(-1) year(-1), respectively. Many semi natural ecosystems in Scotland are vulnerable to nitrogen deposition, including bogs, moorlands, and the woodland ground flora. Because NH3 emissions occur in the rural environment, the local deposition to sensitive ecosystems may be large, making it essential to assess the spatial distribution of NH3 emissions and deposition. A spatial model is applied here to map NH3 emissions and these estimates are applied in atmospheric dispersion and deposition models to estimate atmospheric concentrations of NH3 and NH4+, dry deposition of NH3, and wet deposition of NHx. Although there is a high level of local variability, modelled NH3 concentrations show good agreement with the National Ammonia Monitoring Network, while wet deposition is largest at high altitude sites in the south and west of Scotland. Comparison of the modelled NHx deposition fields with estimated thresholds for environmental effects ("critical loads") shows that thresholds are exceeded across most of lowland Scotland and the Southern Uplands. Only in the cleanest parts of the north and west is nitrogen deposition not a cause for concern. Given that the most intense effects occur within a few kilometres of sources, it is suggested that local spatial abatement policies would be a useful complement to traditional policies that mitigate environmental effects based on emission reduction technologies. PMID- 15349520 TI - The molecular basis for intersexuality part one: the developing testis. PMID- 15349521 TI - Intratesticular adenomatoid tumor radiographically mimicking a primary testicular malignancy. PMID- 15349522 TI - Hematuria due to an intrapelvically intruding total hip replacement. PMID- 15349523 TI - Coexisting angiomyolipoma and renal cell carcinoma in a kidney of an elderly woman: case report and review of the literature. AB - Angiomyolipoma is a well described but relatively uncommon benign renal neoplasm composed of varying admixtures of mature adipose tissue, smooth muscle, and thick walled blood vessels. The incidence of angiomyolipoma is about 0.3% overall. It frequently occurs in patients with tuberous sclerosis. Even more uncommon is the simultaneous occurrence of angiomyolipoma and renal cell cancer in the same kidney in a patient without tuberous sclerosis. PMID- 15349524 TI - Renal trauma: a practical guide to evaluation and management. PMID- 15349525 TI - GU evaluation and management of renal transplant candidates and recipients. AB - There are more than 200,000 end stage renal disease (ESRD) patients who are potential transplant candidates and more than 100,000 previously transplanted renal recipients with functioning allografts in the United States. Fifty-seven percent of these patients are male and forty percent are greater than 50 years of age. Diabetes is the most common cause of kidney failure. It is evident that many patients are at high risk for development of urologic problems and thus it is estimated that the average urologist will care for up to ten of these patients yearly. Thus a review of the genitourinary (GU) evaluation and management of these patients is timely. PMID- 15349526 TI - Bladder outflow obstruction caused by prostatic calculi. PMID- 15349527 TI - Gastrocystoplasty. PMID- 15349528 TI - Urethral foreign body management: a case report. AB - The management of urethral foreign bodies may require the use of various surgical techniques in a urologist's armamentarium. We report a unique case of a urethral foreign body requiring percutaneous and endoscopic techniques for removal. PMID- 15349529 TI - Urinoma: a rare complication of abdominal aortic aneurysm surgery. AB - Urinoma or para-renal pseudocyst generally occurs as a result of trauma to the pelvi-ureteric system. It consists of an encapsulated collection of extravasated urine and is usually located in the peri-renal space or more uncommonly in the peritoneal, pleural or mediastinal cavities. There is only one previously reported case of urinoma secondary to abdominal aortic aneurysm (AAA) surgery. We report a case of symptomatic urinoma after infra-renal AAA repair and discuss the etiology, diagnosis and treatment of this unusual condition. PMID- 15349530 TI - Endourologic management of upper tract transitional cell carcinoma. PMID- 15349531 TI - Urinary tract infections in adults. AB - Urinary tract infection (UTI) is an exceedingly common problem prompting seven million office visits and one million hospitalizations in the United States each year. Advances in the understanding of both host and bacterial factors involved in UTI have led to many improvements in therapy. While there have also been advances in the realm of antimicrobials, there have been numerous problems with multiple drug resistant organisms. Providing economical care while minimizing drug resistance requires appropriate diagnosis, evaluation, and treatment of urinary tract infections. PMID- 15349532 TI - Adenomatous polyp of the verumontanum causing bladder outlet obstruction. PMID- 15349533 TI - Penile fracture and associated urethral injury. PMID- 15349534 TI - Microlaparoscopy in sex reassignment surgery. AB - Sex reassignment (male to female surgery) is a standard operation which is aimed at constructing female genitalia and obtaining a cosmetic and functional result that is similar to that of a normal female subject. The ideal surgical procedure has not yet been described, but the various techniques which have been proposed in the literature are similar. The most cumbersome maneuver of the procedure is that of creating a neovaginal cavity inside the perineum. This step is generally carried out by means of blunt dissection between the rectal wall and the prostate, but most of the surgery is blindly performed without visual control. In these conditions, the risk of rectal injury is high, and may lead to severe intraoperative complications. Microlaparoscopy allows for a direct observation of the perineal dissection from inside the peritoneal cavity, thus avoiding risk of rectal injury. The technique is simple to perform, is non-invasive, and only 15 minutes are added to the operation. PMID- 15349535 TI - Use of the silastic sheath in bladder neck reconstruction. AB - OBJECTIVE: The study compared two populations of patients undergoing bladder neck reconstruction using the silastic sheath in two major pediatric centers. The success with this technique was markedly different in the two centers. The purpose of the study was to determine factors that might explain the divergent results. PATIENTS AND METHODS: Fifteen patients treated in Indianapolis were compared with 94 patients treated in London with the silastic sheath technique of bladder neck reconstruction. Eighty-seven percent of the Indianapolis patients had myelomeningocele whereas 86% of the London group had exstrophy/epispadias. Median age of the Indianapolis patients was 11 years whereas it was 8.4 years in London. Seventy-three percent of patients in Indianapolis were female and 79% in London were male. Patients were followed for a minimum of eight years in Indianapolis and a mean of seven years in London. Similar surgical technique was employed in the two centers but, over time, the London approach included use of a non-reinforced silastic wrapped loosely around the bladder neck with the interposition of omentum. RESULTS: Both groups achieved continence rates exceeding 90%. Of the Indianapolis patients, two-thirds experienced erosion of the silastic at a mean of 48 months. With modifications in the London technique, the erosion rate of silastic was lowered from 100% to 7%. CONCLUSION: Direct, snug wrap of silastic without omentum around the Young-Dees tube as well as simultaneous bladder augmentation placed patients at increased risk for erosion. The silastic sheath technique may be less applicable to myelomeningocele patients. It seems most applicable to older male patients with exstrophy or epispadias undergoing Young-Dees bladder neck reconstruction who have the ability to void. PMID- 15349536 TI - Spinning top urethra and lower urinary tract dysfunction in a young female. AB - Spinning top urethra (STU) denotes a particular urethral configuration that is a dilated posterior urethra mainly seen in young girls or women. STU deformity arises secondary to detrusor instability, leading to a rise the intravesical pressure against a closed sphincter. We describe a case of spinning top urethra in a 30-year-old woman who presented with lower urinary tract symptoms and left flank pain. PMID- 15349537 TI - Renal urologic anomalies presenting in adult identical twins. AB - Two sets of identical adult twins recently presented to our hospital. In one case, the patients demonstrated (ipsilateral) renal agenesis. In the other, the patients presented approximately one year apart with symptomatic (ipsilateral) ureteropelvic junction obstructions. Although the literature suggests a few reports of this type in the pediatric and newborn population, the authors are unaware of similar reports in adults. PMID- 15349538 TI - Physiology of penile erection. PMID- 15349539 TI - Comparison of three types of continent urinary diversions in a single center. AB - The results including the complication and continence rates for 3 types of continent urinary diversion were evaluated. From 1992 to 1998 we performed 58 continent urinary diversions after radical cystectomy for invasive transitional cell carcinoma (TCC) of the urinary bladder. All three types of continent diversions and ileal loop procedures were discussed and patient preferences were determined. The patient preference rate for continent urinary diversion was 96.6%, and half of these patients wanted to be completely dry. Mean age of the patients was 58.2 years. Of the 58 patients, 9 (15.5%) had a Kock pouch, 15 (25.8%) had a Kock neobladder and 34 (58.6%) had sigmoidorectal pouch (Mainz-II pouch). Early and late complication rates of the three different continent diversions were evaluated. The number of complications, such as urine leakage, pyelonephritis, hydronephrosis, reflux and stone formation, were similar in all three types of diversions. Two (5.9%) Mainz pouch II patients who had stopped oral alkalinization demonstrated severe hyperchloremic acidosis. Spontaneous pouch rupture occurred in 1 of the Kock pouches. Reoperation rates were higher with the Kock pouch and Kock neobladder cases. Daytime continence rates for the Kock pouch, Kock neobladder and Mainz II pouch were 77.7%, 86.7% and 100% respectively. Even though complete dryness may not be achieved in every patient, orthotopic bladder substitution appears to be the best choice after radical cystectomy. Although it carries the risk of life-long oral alkalinization therapy, the Mainz pouch II is associated with an excellent continence rate and may be a good alternative for patients who desire to be dry. PMID- 15349540 TI - Testosterone replacement therapy. PMID- 15349541 TI - Epididymal metastasis from carcinoma of the prostate. AB - Epididymal Metastasis from a primary carcinoma of the prostate gland is a rare but recognised phenomena. We describe a case of such metastasis which, unlike previous reports, presents as a painful epididymal mass. Therefore it is important for urologists to consider epididymal metastasis as part of the differential diagnosis in a patient with known carcinoma of the prostate and a tender epididymal mass. PMID- 15349542 TI - Erectile dysfunction under age 40: etiology and role of contributing factors. AB - The aim of this study was to evaluate the etiology of erectile dysfunction (ED) in patients under the age of 40 years. Eighty one patients were included in this study. All patients underwent a multidisciplinary diagnostic approach by color Doppler ultrasonography, dynamic pharmacocavernosometry (optional), selective pudendal pharmaco-arteriography (optional) and nocturnal penile tumescence monitoring by a Rigi-Scan (optional). Mean age of the population was 32 years. Psychogenic impotence was diagnosed in 50% of the patients and organic impotence was diagnosed in 45%. After the 3rd decade of life, a vasculogenic etiology was the most common cause of impotence. Smoking and hypertension played a major role as chronic contributing factors in the overall study population. Primary impotence was diagnosed in 11 patients who were unmarried. The rate of organic causes was 45% in this group (all vasculogenic in nature). Erectile dysfunction in younger patients and in patients with primary impotence is due mainly to organic causes, usually vascular in origin. PMID- 15349543 TI - A rare condition: the ureteritis cystica. PMID- 15349544 TI - Spermatic vein tumor thrombus in renal cell carcinoma. AB - Renal cell carcinoma has the tendency to form venous thrombi. This may involve the renal veins or the inferior vena cava and may extend cephalad/antegrade into the right atrium. We report a patient with renal cell carcinoma who had an intracaval tumor thrombus that had extended into the right spermatic vein. We believe this to be the first description in English literature of a histologically proven renal cell carcinoma thrombus in the spermatic vein. PMID- 15349545 TI - Spontaneous perinephric hemorrhage associated with urolithiasis in pregnancy. AB - Spontaneous perinephric hematoma in the absence of anticoagulation, arteritis, or trauma is uncommon. We report the case of a postpartum patient with nephrolithiasis who initially presented to the obstetric service with a spontaneous perinephric hemorrhage. PMID- 15349546 TI - Laser surgery of the prostate: a review of the current options. PMID- 15349547 TI - Bilateral staghorn calculi in an eighteen-month-old boy. PMID- 15349548 TI - Splenogonadal fusion diagnosed by Doppler ultrasonography. AB - Splenogonadal fusion usually presents as a left scrotal mass but rarely is the diagnosis suspected preoperatively. Herein, we present the first report of a left splenogonadal fusion which was suspected preoperatively by doppler ultrasonography in a 2 year old boy. PMID- 15349549 TI - Pilonidal sinus of the penis. AB - A pilonidal sinus is a subcutaneous sinus containing hair. It is most commonly found in the natal cleft of hirsute men. Here we describe the unusual finding of a pilonidal sinus arising on the male foreskin. PMID- 15349550 TI - Self-retraction clamp for dissection of the posterior prostatic fossa during radical retropubic prostatectomy. PMID- 15349551 TI - Diagnosis, evaluation and treatment of adolescent varicocele. PMID- 15349552 TI - Percutaneous management of urinary calculi. PMID- 15349553 TI - Cowper's syringocele causing painful haematuria in an adult. AB - A Cowper's syringocele in an adult is rare. Ten cases are reported in the world literature. The authors report a case of painful haematuria due to the presence of a Cowper's syringocele in an adult. The classification of lesions of the Cowper's gland is discussed together with common symptoms and differential diagnosis. PMID- 15349554 TI - Renal artery stenosis and ipsilateral renal cell carcinoma: description of an in situ partial nephrectomy and splenorenal arterial bypass. AB - A case of a renal artery stenosis and ipsilateral renal cell carcinoma with long term results is reported. A 65-year-old man with renovascular hypertension, renal insufficiency, and nephrotic range proteinuria presented with an incidental renal cell carcinoma. Concomitant in situ left partial nephrectomy and splenorenal arterial bypass was achieved. The patient is doing well without evidence of malignancy, stable renal function, markedly improved proteinuria and stable blood pressure more than three years later. The techniques of this procedure are detailed and underscore the possibility of successful removal of a renal cell carcinoma with preservation of renal function despite renal artery stenosis. PMID- 15349555 TI - The management of nonseminomatous testicular cancer. PMID- 15349556 TI - Evaluation and management of male urinary incontinence. PMID- 15349557 TI - Superselective embolization of arterial bleeding as a late complication 3 months after nephron sparing surgery for renal cell carcinoma. AB - To our knowledge, this is the first case of an arterial bleeding as a late complication 3 months after nephron sparing surgery of renal cell cancer, presumably originating from an arteriocalyceal fistula. Superselective embolization of the feeding arterial branch was chosen for treatment of the hemorrhage and proved successful. The high efficacy of superselective embolization as a minimally invasive procedure in this and other cases of bleeding Vessels should be the preferred method instead of open surgery. PMID- 15349558 TI - 54 year-old female with recurrent bronchopneumonia and tumor of the left kidney. PMID- 15349559 TI - Incisionless pubovaginal fascial sling using transvaginal bone anchors for the treatment of stress urinary incontinence. AB - INTRODUCTION: Bladder neck suspension (BNS) for stress urinary incontinence (SUI) can have significant morbidity, including bleeding, infection and pain. In an effort to reduce this potential morbidity, we have devised a technique which provides the same suburethral support as a standard anterior vaginal wall sling (AVWS), but without a vaginal or suprapubic incision. We describe this minimally invasive technique. METHODS: From April 1998 to February 1999, 85 women underwent an incisionless suburethral fascial sling procedure. A transvaginal bone drill was used to place a bone anchor loaded with #1 prolene suture into the inferior aspect of the pubic bone on either side of the urethra. A subepithelial tunnel was created at the level of the bladder neck. A 2 x 7 cm segment of cadaveric fascia lata was placed through the subepithelial tunnel. The sutures were passed through the fascia 5mm from either edge, effectively creating a 6.0 cm sling. Finally, the sutures are tied up to the pubic symphysis. RESULTS: Follow-up was via a self-administered questionnaire and patient interview. Recurrent SUI was noted in 2/85 (3%). New onset urge incontinence was present in 4/85 (5%). Permanent urinary retention has not occurred in either group. All procedures were performed on an outpatient basis and no operative complications occurred. CONCLUSIONS: Early results for the incisionless sling compare favorably with the long term results for the AVWS. This minimally invasive approach has thus far not been associated with any significant complications. Elimination of the vaginal and suprapubic incisions has not compromised efficacy, and appears to reduce the incidence of urge incontinence. Long term follow-up will establish the lasting efficacy of this novel surgical technique. PMID- 15349560 TI - Urologic issues during pregnancy. AB - Pregnancy induces a variety of physiologic changes in the urinary tract. When such changes become accentuated the physiologic becomes the pathologic and symptoms arise, at times of significance enough to threaten the well being of mother and/or fetus. This article intends to describe the basis for urinary physiology and its pathologic counterparts during pregnancy. Such a background may then facilitate a rational management protocol for various urologic problems in the gravid state. PMID- 15349561 TI - Radiation therapy after radical prostatectomy. PMID- 15349562 TI - VURD syndrome managed by pyelostomy. AB - We report a case of VURD syndrome in a three day old neonate who was diagnosed with hydronephrosis on a prenatal ultrasound. Severe tortuosity and dilation of the upper urinary tracts in the presence of progression of hydronephrosis or a persistently elevated creatinine may favor a proximal urinary diversion rather than primary valve ablation or cutaneous vesicostomy. Because of a persistently elevated serum creatinine, a nonfunctioning kidney with grade 4/5 vesicoureteral reflux and worsening contralateral hydronephrosis despite lower tract drainage, a left cutaneous pyelostomy was performed, contralateral to the kidney involved with VURD. Postoperatively the serum creatinine stabilized at 1.0 mg/dl and decreased to 0.3 mg/dl at one month of age. PMID- 15349563 TI - Superficial bladder cancer therapy. AB - Bladder cancer treatment remains a challenge despite significant improvements in preventing disease progression and improving survival. Intravesical therapy has been used in the management of superficial transitional cell carcinoma (TCC) of the urinary bladder (i.e. Ta, T1, and carcinoma in situ) with specific objectives which include treating existing or residual tumor, preventing recurrence of tumor, preventing disease progression, and prolonging survival. The initial clinical stage and grade remain the main determinant factors in survival regardless of the treatment. Prostatic urethral mucosal involvement with bladder cancer can be effectively treated with Bacillus Calmette-Guerin (BCG) intravesical immunotherapy. Intravesical chemotherapy reduces short-term tumor recurrence by about 20%, and long-term recurrence by about 7%, but has not reduced progression or mortality. Presently, BCG immunotherapy remains the most effective treatment and prophylaxis for TCC (Ta, T1, CIS) and reduces tumor recurrence, disease progression, and mortality. Interferons, Keyhole-limpet hemocyanin (KLH), bropirimine and Photofrin-Photodynamic Therapy (PDT) are under investigation in the management of TCC and early results are encouraging. This review highlights and summarizes the recent advances in therapy for superficial TCC. PMID- 15349564 TI - Surgery of the adrenals. PMID- 15349565 TI - Urologic problems in patients with acquired immunodeficiency syndrome. PMID- 15349566 TI - Uretero-infundibuloplasty for giant hydronephrosis. PMID- 15349653 TI - [The measurement parameters and statistical analysis of therapeutic study]. AB - We can draw a correct conclusion unless we select appropriate measurement parameters and properly do the statistical analysis including description and inference, after checking and processing the data of therapeutic study. A comprehensive understanding and correct application of the measurement parameters as well as statistical processing is the key to improve our scientific research. In this article, some common mistakes were addressed in the light of basic concepts in therapeutic study, focusing on the measurement parameters (survival rate, negative conversion rate or positive conversion rate, relative benefit increase, relative risk reduction, relative risk increase, number needed to treat, likelihood of being helped vs. harmed, index of effectiveness) and statistical processing (data type, distribution character, intention-to-treat analysis, efficacy analysis, treatment received analysis, hypothesis testing). PMID- 15349654 TI - [A preliminary evaluation of alleviating damage due to scars postoperative in the area of the parotid gland]. AB - To estimate the methods of alleviating damage due to postoperative scars in the area of the parotid gland. 12 clinical cases was chosen which accepted operations in the area of the parotid gland, the method of minimally invasive suture was used in the operation and the remedy which can inhibit the hyperplasia of the scars was used after operation. 8 cases were without conspicuous scars, 4 had mild scars, no case was found with hyperplasia of scars. The influence of facial contour due to postoperative scars can be controlled by the above stated methods. PMID- 15349655 TI - [Relationship between condylar marrow signal abnormalities and temporomandibular joint pain]. AB - PURPOSE: To investigate the relationship between condylar marrow signal abnormalities and joint pain. METHODS: Oblique sagittal T1 and T2 weighted MR imaging at closed mouth was obtained from 88 joints of 44 patients who complained of unilateral TMJ pain. The condylar marrow signal of pain-free side served as self-control. All patients rated their pain levels by a visual analogue scale (VAS). RESULTS: Of 44 painful joints, 11(25% joints showed condylar marrow signal abnormalities, all of which were edema pattern. While there had condylar marrow signal abnormalities only in 2 (4.5%) of 44 pain-free TMJs. There was significant correlation between joint pain and condylar marrow signal abnormalities (P<0.01). The VAS score of patients with and without condylar marrow signal abnormalities was respectively 39.5+/-27.5 and 42.6+/-21.9, There was no correlation between them (P=0.696). CONCLUSION: Temporomandibular joint pain is closely correlated with condylar marrow signal abnormalities, but the pain degree has no association with it. PMID- 15349656 TI - [The effect of recombinant human erythropoietin on postoperative anemia in orthognathic patients]. AB - PURPOSE: To observe the effect of rhEPO on postoperative anemia in orthognathic patients. METHODS: 31 patients had 500-1200 ml blood loss during orthognathic operation, who were divided into two groups randomly, the experiment group and the control group. The patients in the experiment group received rhEPO 6000IU subcutaneously for 3 times in a week and Ferrost sulfatis Et vitamini-medtech 1 tablet per day via oral administration for 12 days after operation, while the patients in the control group only received the same dose of Ferrost sulfatis Et vitamini-medtech. The loss of blood of the patients during operation were estimated and recorded. Their Hb and Hct were mensured before operation, and in the first, third, seventh-twelfth day after operation. RESULTS: In the twelfth day after operation, the Hb and Hct of the patients in the experiment group had a significantly higher increase than that in the control group (P<0.05). CONCLUSIONS: rhEPO combined Ferrost sulfatis Et vitamini-medtech are suitable to apply in orthognathic patients with anemia after operation, which may effectively accelerate the recovery of their anemia, and avoid transfusion of blood. PMID- 15349657 TI - [The clinical study on the adjunctive effects of aqueous extract from coptis root for the treatment of chronic periodontitis]. AB - PURPOSE: To make a therapeutic membrane with aqueous extract from coptis root and explore its adjunctive effects for treating chronic periodontitis. METHODS: Drug membrane from coptis root aqueous extract was developed; 4 teeth in 30 patients with moderate to advanced periodontitis were randomly divided into four groups: coptis root membrane, iodine glycerin, single drug membrane and blank control group. All parameters including plaque index (PI), probing depth (PD), attachment loss (AL) and bleeding on probing (BOP) were measured at baseline, 4 and 7 weeks after treatment. Analysis of variance and chi-square test were carried out for analysis. RESULTS: In all four groups, there were significant differences of PD, AL, BOP between baseline and 4,7 weeks after treatment (P<0.05), the treatment effect of coptis root membrane was significantly superior to that of other three groups (P<0.05). Moreover, all the parameters improved continuously. CONCLUSION: Use of coptis root membrane as an adjunctive method after scaling can significantly improve the treatment effect of periodontitis. PMID- 15349658 TI - [The inhibitive effect of IgY toothpaste against oral Streptococcus mutans]. AB - PURPOSE: To study the effect of the toothpaste containing IgY against oral Streptococcus mutans. METHODS: 140 subjects were divided into 2 groups randomly (test group and control group). Before the beginning of the test, every subject underwent a elution period in order to lessen the other interference factors, then used different toothpastes were used respectively and the test began. Streptococcus mutans values were determined with Dentocult SM Strip just before test, at 1, 3, 7, 30 days after toothpaste used and 14 days after stopping toothpaste used. RESULTS: (1) In test group,Streptococcus mutans values decreased at 1 day after toothpaste used. In control group Streptococcus mutans values decreased at 3 days after toothpaste used. (2) Streptococcus mutans values decreased gradually in each groups after toothpaste used. (3) In test group, 2 weeks after stopping toothpaste used, Streptococcus mutans values were still suppressed. CONCLUSION: The application of the toothpaste containing IgY may reduce oral Streptococcus mutans levels significantly. PMID- 15349659 TI - [The change of F2 and B2 in patients with complete denture]. AB - PURPOSE: To study the clinic adaptive effect of speech articulation of patient with complete denture. METHODS: The sensitive utterance study was /zi/, /ci/, /si/, which was input into CSL directly before and after patient's restoration at the beginning, then at 1 week, 2 weeks, 4 weeks and 8 weeks, B2 of the utterance of 30 edentulous patients were measured by the computer. RESULTS: There were significant differences in F2 and B2 before and after patient's restoration. CONCLUSION: There is a definite regularity of speech articulation after restored with complete denture. CSL helps us to analyze speech articulation by visual accessing and provides basic theory of evaluation in clinical dental prosthetics. PMID- 15349660 TI - [The effects of combined reverse Nance and TPA on reinforcement of anchorage]. AB - PURPOSE: To evaluate the effects of combined reverse Nance and TPA on anchorage control. METHODS: 16 female adults with Class II division 1 malocclusion were treated with the combined mechanism of reverse Nance and TPA in an attempt to reinforce molar anchorage. Cephalometric analysis and model measurements were conducted for the assessment of anchorage control. RESULTS: The mesial movement of the upper first molars were minimized within 1/5 extracted space, parameters such as UMVD, MP-FH, ANS-Me remained unchanged, indicating the molar anchorage was controlled effectively. CONCLUSION: The modified combination of reverse Nance and TPA might be effective in anchorage forcement. PMID- 15349661 TI - [The influence of cerebral blood flow of volunteers with magnetic attachment measured by transcranial Doppler]. AB - PURPOSE: To investigate the influence of cerebral blood flow of volunteers with magnetic attachment. METHOD: The mean flow velocity of the middle cerebral arteries (MCA) of 24 volunteers with magnetic attachment denture were measured by transcranial Doppler (TCD). All data were analyzed by SPSS10.0. RESULTS: The results showed the mean flow velocity of MCA of 24 volunteers with magnetic attachment was not significantly different from those without magnetic attachment (P>0.05). CONCLUSION: The magnetic attachment had little effect on cerebral blood flow. PMID- 15349662 TI - [Comparative study of craniofacial structure between UCLP and Class III malocclusions]. AB - PURPOSE: The aim of this study was to compare the dentocraniofacial morphology between the patients with cleft lip and palate and those with Class III malocclusion without surgical treatment. METHODS: The sample included two groups, group one consisted of 39 patients with repaired completed or uncompleted unilateral cleft lip and palate, with age ranging from 12 to 15 years; group two consisted of 40 patients with Class III malocclusion without surgical treatment, with age ranging from 12 to 15 years. Cephalometrics analysis was conducted to allow for a quantitative assessment of dentocraniofacial morphology. RESULTS: The group with unilateral cleft lip and palate showed smaller SNA and SNB angle than those in the Class III group,but the ANB angle showed no significant difference between two groups. Cleft group showed significant downward and backward rotation of the mandible associated with a more obtuse gonial angle. The upper incisors in cleft group were also found to be more retroclined. The inclination of lower anterior teeth showed no difference between two groups. CONCLUSION: The cleft group showed smaller maxilla and mandible than the Class III group, with mandibular plane being clockwise rotated. PMID- 15349663 TI - [Prognosis of three-dimension change at middle facial soft tissue with distraction osteogenesis]. AB - PURPOSE: To evaluate the changes in middle facial soft tissue caused by distraction osteogenesis. METHODS: A three-dimensional finite element model was established based on the simulation of a 13-year-old female with skeletal class III malocclusion. Three approaches of osteotomy, namely Le Fort I, Le Fort II and Le Fort III, were simulated and the maxillary complex was advanced by 5mm in the direction of FH plane, functional occlusal plane and functional occlusal plane with deviation of counter-clockwise 10 degrees, respectively, some premeters indicating the contour of the facial soft tissue were evaluated. RESULTS: The three kinds of osteotomy in which different levels of osteotomied maxillary complex area were advanced. Changing in detail is that the nose tip moved forward 1.25 mm, upper lip moved forward 4.9 mm, and the angle of Cm-Sn-U1 tended to increase after Le Fort I osteotomy; and nose tip moved forward 4.9 mm after Le Fort II osteotomy without increase of the angle of Cm-Sn-U1; besides changes that happened after Le Fort II osteotomy, lower eyelid moved forward 2mm after Le Fort III osteotomy. CONCLUSION: By computer surgery simulation using three-dimensional finite element model, we can obtain not only the amount of the three-dimensional change of the facial soft tissue, but also the three-dimensional effected graphics after operations. PMID- 15349664 TI - [The effect of static magnetic field on bone morphogenetic protein-2 in periodontal membrane of the rat]. AB - PURPOSE: The purpose of this study is to evaluate the effect of static magnetic field on the expression of BMP-2 in the periodontal membrane of rat. METHODS: 35 female rats of wistar were randomly divided into three groups, which were the normal group, experimental control group and experimental group. The experimental rat's cheeks were put into magnet with the intensity of magnetic field being 0.12 tesla. The rats were sacrificed at 2 days, 4 days, 7 days, respectively. Immunohistochemical assays were used to evaluate the changes of BMP-2 expression in the periodontal membrane of the maxillary molars. RESULTS: BMP-2 was mainly found in the plasma of fibroblast, osteoblast, cementocyte, odontoblast separately. There were more BMP-2 in the experimental group treated with static magnetic field. There were significant differences between the experiment group of 4 day and 7 day and the control group. CONCLUSIONS: Static magnetic field stimulated BMP-2 secreted by the cell of periodontal membrane and played an important role in repairing and remodeling of periodontitis. PMID- 15349665 TI - [Bone formation by seeding bone marrow stromal cells on the sodium calcium phosphate/beta tricalcium phosphate scaffold]. AB - PURPOSE: To observe the osteogenesis of sodium calcium phosphate/beta-tricalcium phosphate scaffold loaded with marrow stromal cells in a nude mice subcutaneous model. METHODS: To transform the sintered bovine cancellous bone into NaCaPO(4)/beta-TCP biphasic calcium phosphate ceramic by physical and chemical methods. Bone marrow stromal cells (BMSCs) were obtained by harvesting the cancellous bone from the iliac crest of a rabbit. Then the BMSCs were isolated, expanded and induced in vitro and seeded in the NaCaPO(4)/beta-TCP ceramic scaffolds. The scaffold/BMSCs composites were implanted subcutaneously into the backs of nude mice. Implantation of NaCaPO(4)/beta-TCP ceramic alone was acted as control. At 4 and 8 weeks after implantation, the specimens were harvested and the osteogenetic activity were evaluated by gross and histologic observation. RESULTS: At 4 weeks after implantation, the relatively mature bone were seen on the surface of the material, the inner part of the material were mainly cartilaginous bone. At 8 weeks after implantation, A large amount of trabecula bone had formed and the medullary cavity, medullary cells and lipocytes could be seen. Osteoblasts and osteoclasts were seen between the scaffolds and the local bone in the transition area. The endochondral bone formation pattern were observed. CONCLUSIONS: The NaCaPO(4)/beta-TCP scaffold/BMSCs composites showed good osteogenetic activity and could promote mineralization of the immature bone. It can be used as the bone tissue engineering scaffolds. PMID- 15349666 TI - [Reconstruction of mandibular defect by using Titanium mandibular prosthesis: a preliminary experimental study]. AB - PURPOSE: The aim of this experiment was to investigate the possibility of application of mandibular reconstruction with titanium mandibular prosthesis. METHODS: Two healthy adult goats were used in this study. Spiral CT of the mandibular region was performed. The data collected was related to the computer. Then a 5 cm mandibular body defect and a mandibular angle defect were designed accordingly. Titanium mandibular prostheses were manufactured according to the size of the above defect. Surgical operation was performed and the prosthesis were implanted into the aimed defects and internal fixation was used. Autogenic bone graft was introduced into the holes of the angle prosthesis. Radiographic examination was used to evaluate the healing of prostheses every month after operation. Three months later, the animal was sacrificed and the mandible specimens were sectioned for histological and biomechanical evaluation. RESULTS: One of the body prosthesis was found to be exposed two weeks after operation. Four weeks later, fixation failed and the anterior part of the prosthesis separated from the mandible attached. The other body prosthesis also suffered from exposure,but the connection between the prosthesis and the mandible was still stable. Histological examination revealed that there was presence of osseointegration in the angle prosthesis as well as area that was not exposed in the body prosthesis model. Reattachment of the muscles to the bone graft area was found. CONCLUSIONS: CAD/CAM generated Titanium mandibular prosthesis is applicable in human model. The vitality of this experiment revealed that a tight closure plays an important role in the healing by osseointegration. If there was any exposure, the osseointegration will be failed. Reattachment of the muscle to the prosthesis can be achieved by "muscle-bone-prosthesis"model. PMID- 15349667 TI - [The long-term effects of rhBMP-2 and rhbFGF on alkaline phosphatase of bone mesenchymal stem cells]. AB - PURPOSE: To evaluate the long-term effects of rhBMP-2 and rhbFGF on alkaline phosphatase(ALP) of rabbit bone mesenchymal stem cells by using them alone, associatedly and sequentially. METHODS: Under cell culture technique, ALP kits were used to evaluate ALP level of bone mesenchymal stem cells so as to reflect differentiation of the cells on the 2nd, 5th, 7th, and 10th day by adding rhBMP-2 and rhbFGF on different ways. RESULTS: Statistical analysis showed that rhBMP-2 could enhance while rhbFGF could inhibit the ALP level of rabbit bone mesenchymal stem cells significantly in long term, and also in a dose-dependent manner. The effects of using rhBMP-2 alone was similar to that of using the two growth factors associatedly, and they were significantly higher than that of using rhbFGF alone and using them sequentially. CONCLUSION: Different growth factor has different long-term effect on differentiation of bone mesenchymal stem cells by adding them on different ways. PMID- 15349668 TI - [The effect of disinfection on dimension stability of impressions]. AB - PURPOSE: To measure the effect of disinfection on dimension stability of alginate, hydrocolloid, and Express putty impressions which were separately immersed in 2% glutaraldehyde solution and 1% 84 disinfector. METHODS: Alginate, hydrocolloid, and Express putty impressions were made by 20 mm x 20 mm x 40 mm standards test wares. There were 45 in each kind of test wares. They were divided into 9 groups and 5 pieces in every group. Among them, one group using clear water to flush acted as the matched control, the other groups received respectively 2% glutaraldehyde solution and 1% 84 disinfector immersed in 10 minutes, 20 minutes, 30 minutes. After two hours, the spiral instrument was used to measure the model. The experiment data were analyzed by using the SPSS 11.0 statistic software package. RESULTS: There was no significant difference in pure water flush and spraying treatment respectively. There was significant difference for alginate and hydrocolloid impression was immersed before and after. There was no significant difference for Express putty silicon rubber impression materials before and after immersion. There was no significant difference in 2% glutaraldehyde solution and 1% 84 disinfector when impressions were immersed in. CONCLUSION: Alginate, hydrocolloid and Express putty impressions immersed in 2% glutaraldehyde solution and 1% 84 disinfector separately for 20 minutes does not affect the accuracy, and can be used clinically. PMID- 15349669 TI - [The comparative study of the effect of silica and calcium carbonate based dentifrice on dentine abrasives in vitro]. AB - PURPOSE: To evaluate the effect of Silica (SiO(2)) and Calcium Carbonate (CaCO(3)) based dentifrice on their dentine abrasives. METHODS: Microscope, environmental electron scanning electron microscope (SEM) and radioactive Dentine Abrasion (RDA) assay were used to quantitatively measure the wear of dentine. RESULTS: The mechanical brushing assay showed severe dentine abrasion with CaCO(3) based dentifrices. There were only minor scratches on the specimens brushed with SiO(2) dentifrices. Analysis of the thin section by SEM after exposure to dentifrices showed that NaF/SiO(2) produced significant remineralization whereas MFP/CaCO(3) system resulted in little remineralization. RDA values were 86.17 and 98.61 for SiO(2) based dentifrices and 222.85 and 253.84 for CaCO(3) based dentifrices, respectively. The morphology of the irregular shaped CaCO(3) with sharp points/edges further explained the brushing and RDA results. CONCLUSION: These results indicate that abrasive in dentifrice, e.g. CaCO(3), can cause severe abrasion of dentine in vitro. PMID- 15349670 TI - [Clinical observation on reimplanting the upper inversely impacted incisor after extraction]. AB - In order to determine the feasibility of reimplantation of upper inversely impacted incisor, we enlarged the interdental distance by orthodontic appliance, then replanted the impacted incisor after extraction. 13 of 15 cases had good results. It was a good method to replant inverse impacted incisor after extraction which avoid extracting the tooth simply. PMID- 15349671 TI - [The biological effect of retinoic acid on cultured human periodontal ligament cells]. AB - PURPOSE: To evaluate the biological effect of retinoic acid on cultured human periodontal ligament cells. METHODS: Human periodontal ligament cells were cultured and challenged by retinoic acid with different concentrations. The cell morphology was observed under phase-contrast microscope; the cell proliferation activity was examined using MTT; the alkaline phosphatase activity was measured by enzyme kinetics method; the total amount of protein was assayed at different times. RESULTS: Retinoic acid enhanced alkaline phosphatase activity and had no effect on morphology, proliferation and total protein synthesis of periodontal ligament cells. CONCLUSION: Retinoic acid improves periodontal ligament cells to differentiate into osteoblast like cells. PMID- 15349672 TI - [Study of the anatomical location of hypoglossal nerve at tongue base of male adults]. AB - PURPOSE: This study was undertaken to analyze the anatomical location of hypoglossal nerve at tongue base of male adults to avoid creating a function deficit during radiofrequency of tongue base in OSAHS. METHODS: Ten tongues of male adults in frozen were coronally dissected at foramen cecum, 5 mm,10 mm, 15 mm before/after the foramen cecum. Vertical and horizontal distance of hypoglossal nerve to the surface and middle line of the tongue were measured using computer image procession in micrometer. The data were analyzed with such statistical methods as descriptive statistics, paired t-testing, linear correlation analysis. RESULTS: (1) The anatomical location of the bi-lateral hypoglossal nerve is symmetrical in principle. (2) At the tongue base: The distance of hypoglossal nerve to the surface is 22.21+/-2.22 mm, and has no correlation to the length and the width of the tongue. There are some regularities of the anatomical location of the hypoglossal nerve before/after the foramen cecum. Before the foramen cecum the hypoglossal nerve is located in the mid-medial (21.89+/-1.93)% and after the foramen cecum is in the mid-medial (28.61+/-2.66)%. CONCLUSION: We conclude that if the depth of the needle-polar penetrating into the tongue base doesn't exceed (22.21+/-2.22)mm, and keep away from mid-medial (21.89+/-1.93)% ( before the foramen cecum ) or mid-medial (28.61+/-2.66)% after the foramen cecum, it may be helpful to avoid creating a function deficit when radiofrequency tongue base in snoring and OSAHS is performed. PMID- 15349673 TI - [The effect of mold temperature on the accuracy of titanium castings]. AB - PURPOSE: To study the effect of mold temperature on the accuracy of titanium castings invested in the ZrP99 investment. METHODS: 30 artificial stone dies simulated the post-tooth and their corresponding complete crown wax patterns were made with a truncated cone-shape standard brass model with round sleeves. They were divided randomly into five groups and each group had six samples. Four group wax patterns were invested in ZrP99 investment as test groups and one group was invested in Titavest-CB investment as control. The test group moulds were cast at the mould temperature 600 degrees centigrade, 450 degrees centigrade, 300 degrees centigrade, 150 degrees centigrade, respectively. Measure the distance separating the gingival margin of the crown and the shoulder of the die in the same way as for the wax pattern. The casting accuracy was represented by the discrepancies derived from the space distance measurements made with wax patterns and cast crowns. t test and one-way ANOVA were used for statistical analysis. RESULTS: Measurement of the casting accuracy indicated that the casting accuracy of the control group was higher than all the test groups. When the mold temperature was lower than 450 degrees centigrade, the accuracy of the castings invested in ZrP99 investment was improved significantly with the mold temperature raised. CONCLUSION: To satisfy the requirement of the clinical practice in the point of the casting accuracy, the mold temperature of ZrP99 investment must be chosen higher than 300 degrees centigrade. PMID- 15349674 TI - [The experimental study of the relationship between preparation height and retention of complete crowns]. AB - PURPOSE: This study was to discuss the influence of preparation height on retention of complete crown. METHODS: In this study we measured the retention of complete crown with the preparation height 2 mm, 3 mm, 4 mm and 5 mm. All crowns had no preparation taper and no bonding agent was used. The retention of crowns was measured and the data was statistically analyzed by t test. RESULTS: The preparation height was directly proportional to the retention of complete crown. And the crown with the height of 2 mm had enough retention when the crown had no preparation taper. CONCLUSION: In order to get enough retention of short height molar, we should induce the preparation taper. PMID- 15349675 TI - [Histological changes on the stress side in orthodontic rapid tooth movement through distraction osteogenesis of the periodental ligament on dogs]. AB - PURPOSE: To study histological changes under the conditions of orthodontic rapid tooth movement through distraction osteogenesis of the periodental ligament on dogs. METHODS: The experiment was carried out in 6 dogs, the left side of jaws of each one was set as test or control side, and the other side was control or test side. On the control side, the first premolar was moved using traditional methods while the third premolar as anchor, on the test side, using self-made distraction device. The periodental tissue of tooth moved were extracted at the end of the test, some of decalcified sections were stained with hematoxylin and eosin and others with modified Mallory's trichrome staining method, being examined by LM. RESULTS: Decalcified sections stained with hematoxylin and eosin showed the bone formed actively, and there were a large number of fibroblasts and osteoblasts as well as abundant vascularity. The modified Mallory's trichrome staining method showed the newly formed bone very clearly and distinctly. CONCLUSIONS: There was no difference in quality but in quantity on the histological reactions in tension side of the tooth moved by traditional method and by distraction osteogenesis through the peridental ligament and periodontal membrane, the latter could induce higher activity of histological synthesization than the former. PMID- 15349676 TI - [Survival analysis--the method used to analyze the follow-up data]. AB - In clinical research, the collected data covers a kind of special data that named follow-up data. However, long time's follow-up is necessary when to evaluate the effect of certain drugs or operations on the patients. During this process, various kinds of outcome and different follow-up times will be found among patients. Survival analysis is a kind of statistical methods that connect outcome with survival time and co-analyze them. This statistical method is used to analyze completed data and uncompleted data at the same time, which can make the best use of the information in medical research to evaluate and compare the follow-up data more exactly. This review introduces the basic principle and method of survival analysis. PMID- 15349677 TI - [Defensins and tumor immunity and its relationship to oral tumors]. AB - Defensins play a vital part in immunodefensity, with broad-spectrum of antimicrobial activities. Defensins can kill G(+), G(-) bacterial, fungal and some virus. At the same time, their cytotoxicity and cytolysis in tumor immunity are getting more and more attention. At present, studies on defensins in tumors especially in oral tumors increase. This paper summarizes defensins and tumor immunity and its relationship to oral tumors. PMID- 15349678 TI - [Advances in the research of all-ceramic FPD]. AB - The all-ceramic restoration has no metal base,no need to mask, and it also has perfect esthetic attribute. But in the past, lower strength restricted its clinical use. With the development of high strength ceramics, all-ceramic restoration has been widely used than before. Recently, more and more research on all-ceramic materials, laboratory test and clinical test showed good prospect in prosthodontics field. PMID- 15349679 TI - [Combined orthodontic and orthognathic therapy on skeletal class II division 1 malocclusion in adults]. AB - PURPOSE: To Introduce a combined orthodontic and orthognathic method for treatment of skeletal class II division 1 malocclusion in adults. METHODS: In orthodontic-surgical treatment on skeletal class II division 1 malocclusion in adults, the goal of orthodontics was to remove the compensation of upper and lower anterior teeth, leveling Spee's curve, adjusting relationship of the upper and lower dentition arch for intercuspid bite after surgery. The goal of orthodontics before surgery was visual treatment objective (OTV), model surgery and making of bite-plate. To achieve these goals, a combined orthodontic and orthognathic method for treatment of skeletal class II division 1 malocclusion in adults was adopted in this study. RESULTS: Stable function of stomotognathicia and satisfactory facial aesthetics after treatment were obtained. CONCLUSION: Oral and maxillofacial deformity must be treated by combining orthodontic treatment and surgical osteotomy in order to restore oral function and aesthetic appearance. PMID- 15349680 TI - [Eosinophilic granuloma of the jaw: an analysis of 21 cases]. AB - PURPOSE: To study the clinical features and treatment of eosinophilic granuloma of the jaw (EGJ) in 21 cases. METHODS: 21 patients with EGJ treated from 1983 to 2002 were reviewed, including the sexes, ages, extent of lesions, clinical features and treatment methods. RESULTS: The male to female rate was 13:8. 76% of the cases were among 2-10 years. The median age was 8 years, 18 lesions were in the mandible, 1 was in the maxilla and 2 involved the mandible and maxilla. CONCLUSIONS: EGJ was rarely seen clinically, lack of specificity. Pathology can confirm the diagnosis. Surgery is still the major treatment modality. Combination of radiotherapy or chemotherapy maybe valuable. The prognosis of the patients was generally good. PMID- 15349681 TI - [Study on the characteristics and reasons for mistakes in some orthodontic cases]. AB - PURPOSE: To evaluate the characteristics and reasons for mistakes in some orthodontic cases. METHODS: 37 cases after or during treatment in other hospitals with mis-treatment were chosen, their original records as well as the records when they came to our hospital were collected, their original and present malocclusions were classified and the characteristics and reason of mistakes were analysed. RESULTS: Among the 37 cases, 26 cases had class-III malocclusion which account for 70.27% of the group, especially for the malocclusions began in mixed dentition and the problems developed from anterior dental crossbite to severe skeletal crossbite with dentofacial asymmetry and openbite. The main reasons lie in the mistakes of diagnosis and treatment planning, preventive treatment,appliance fabrication,use of extraoral arch and elastics, treatment processes, et al. CONCLUSIONS: There are various characteristics and reasons of treatment mistakes, but the problems lie mainly in class-III malocclusion especially during mixed dentition which indicates that we should pay more attention to the treatment of class-III malocclusion. PMID- 15349682 TI - [Observation of the change of blood pressure in 835 patients during extraction of teeth]. AB - PURPOSE: To observe the change of blood pressure during extraction of teeth. METHODS: Patients without hypertension who needed extraction were randomly selected, tooth extraction was carried out. The blood pressure and the patient's response to operation were observed and recorded during the procedure. RESULTS: The blood pressure in 825 patients increased significantly. The average increased blood pressure was 45/26 mmHg, 3 patients had arrhythmia, 5 patients had adverse reaction. CONCLUSION: The patient's blood pressure must be fewer than 140/90 mmHg, before a difficult extraction of teeth. PMID- 15349683 TI - [A study of clinical effects on treatment of 225 cracked-teeth]. AB - PURPOSE: To analyse the clinical effects of cracked-teeth after filling or full crown restoration. METHODS: 225 cracked-teeth were divided into two groups: the early 77 cracked-teeth and the late 144 cracked-teeth according to their clinical features and diagnosis, and then different interventions were applied, the clinical effects were recordered and analysed. RESULTS: There was a significant difference after the early 77 cracked-teeth being treated by filling or full crown rehabilitation after 3 years follow up. The treatment to the late 144 cracked-teeth was excellent after 3 years' observation with root canal therapy or pulp mummification, and finally full crown restoration. CONCLUSIONS: By diving cracked-teeth into different stages and giving corresponding treatment, the cracked-teeth can be preserved successfully. PMID- 15349684 TI - [Application of saucer shaped cavity and retentive groves in class II cavity restoration]. AB - PURPOSE: To observe the clinical effects of class II cavity restoration by using saucer shaped cavity and retentive grooves and dovetail retentive cavity. METHODS: 550 cases needing to have class II cavity preparations were divided into two groups randomly to have class II cavity restoration by using saucer shaped cavity and retentive groups or dovetail retentive cavity. The same light cured posterior composite resin restorative material was used 2 groups. After two years of follow-up, The clinical effects were recorded and evaluated. RESULTS: There was no significant difference in two groups (P>0.05) in respects to loss of retention, margin sealing, Secondary caries, grooves occlusal abrasion, pulp reaction and margin coloration. CONCLUSION: Using saucer shaped cavity and retentive grooves had the similar effect to dovetail retentive cavity. PMID- 15349685 TI - [Treatment of gingival recessions of teeth with ceramic full crown by autogenous connective tissue graft]. AB - PURPOSE: The aim of this clinical study was to evaluate gingival recession of teeth with ceramic full crown treated with subepithelial connective tissue grafts. METHODS: In 20 systemically healthy, whose ages ranging from 31 to 58 years, 32 teeth with ceramic full crown developed gingival recession. Selection of gingival recession defects was based upon the following criteria: A minimum of 2 mm of root surface exposure in coronal-apical height. Miller class I and II recession. Interproximal and labial probing depth time in tutorials > time spent on dissection). For the American model (also in the UK and some other parts of Europe), the training is often characterized by being dissection-based (time spent on dissection > time in lectures > time in tutorials; 10 medical schools surveyed). Exceptionally, in one Australian school, time in tutorials exceeds time in lectures (dissection = 0). The differences between the French and American models relate to teaching aims-where dissection predominates, the aims are not just the learning of anatomical facts but include practical skill acquisition and experiential learning. In 2001, to help us change the methods of teaching of anatomy in our medical school at CHU Necker-Enfants Malades (Paris V, France), we asked other French medical schools (and some foreign schools) to suggest ways of organizing anatomy training within certain time constraints. In this paper, we present the answers received. The responses received were of two kinds: (1) those providing a description of the anatomy teaching in their own medical school; (2) those providing a system for organizing the teaching if we, in Paris, have 120 hours in total to teach gross anatomy (except neuroanatomy). In the latter case, a considerable variety of different, and innovative, alternative schemes were suggested that are described in this article. PMID- 15349696 TI - Volume variation of mastoid pneumatization in different age groups: a study by three-dimensional reconstruction based on computed tomography images. AB - Although there have been some reports that measured the size of mastoid pneumatization, only a few studies have reported the age-related variations in the mastoid air cell system using three-dimensional (3D) reconstruction techniques of computed tomography (CT) images. We performed a retrospective, cross-sectional study. A 3D reconstruction based on CT images was performed on 199 ears of 102 patients (age range 6-84 years) without otologic disease by a surface-rendering algorithm. The results showed that mastoid pneumatization continued to grow until the third decade. Thereafter, it declined slowly, and then rapidly after the seventh decade. No statistically significant difference was found between male and female or between right and left sides. There was a significant difference between the larger and smaller sides of individuals. The volume measurement technique based on the 3D reconstruction technique reported here is widely available, highly accurate and easy to perform. PMID- 15349698 TI - Modification of humic acids by the compost-dwelling deuteromycete Paecilomyces inflatus. AB - The soil mold Paecilomyces inflatus is capable of modifying and partially mineralizing synthetic and natural humic acids (HAs) in compost environments. HA degradation studies using a synthetic HA (14C-HA) in autoclaved compost microcosms showed that, after 12 weeks of cultivation, P. inflatus mineralized approximately 5% of the 14C-labeled HA to 14CO2, while 6% of the 14C-HA was converted into 14C-labeled water-soluble fragments (fulvic-acid-like fraction). About 40% was still present as NaOH-soluble HA representing unmodified or only slightly modified humic material (compared with 60% in the controls). Modification of natural HAs extracted from compost was followed by their partial decolorization (30%) in liquid cultures of P. inflatus. Bleaching of the medium was accompanied by moderate changes in the molecular mass distribution of both the HA and fulvic-acid fractions, which were analyzed with high-performance size exclusion chromatography. HA modification was most pronounced during the primary growth phase of the fungus and was associated with increased laccase activity. PMID- 15349699 TI - Expression of laccase IIIb from the white-rot fungus Trametes versicolor in the yeast Yarrowia lipolytica for environmental applications. AB - Improvement of the catalytic properties of fungal laccases is a current challenge for the efficient bioremediation of natural media polluted by xenobiotics. We developed the heterologous expression of a laccase from the white-rot fungus Trametes versicolor in the yeast Yarrowia lipolytica as a first step for enzyme evolution. The full-length cDNA consisted of a 1,561-bp open reading frame encoding lacIIIb, a 499-amino-acid protein and a 21-amino-acid signal peptide. Native and yeast secretion signals were used to direct the secretion of the enzyme, with the native signal yielding higher enzyme activity in the culture medium. The level of laccase activity secreted by the transformed yeast was similar to that observed for the non-induced wild-type strain of T. versicolor. The identity of the recombinant enzyme was checked by Western blot and matrix assisted laser desorption/ionization time-of-flight analysis. Electrophoresis separation in native conditions indicated a molecular mass of the recombinant protein slightly higher (5 kDa) than that of the mature T. versicolor laccase IIIb, suggesting a limited excess of glycosylation. The laccase production level reached 2.5 mg/l (0.23 units/ml), which is suitable for engineering purpose. PMID- 15349700 TI - Chemo-enzymatic synthesis of 2'-O-methoxyethyl ribonucleosides using a phosphodiesterase from Serratia marcescens. AB - An enzyme able to cleave the 3',5'-phosphate ring of 2'-methoxyethyl cyclic nucleotides (3',5'-cyclic nucleotide phosphodiesterase, EC 3.1.4.17) from Serratia marcescens DSM 30121 was used to deprotect the cyclic phosphate nucleotides after chemical alkylation. The process yielded 2'-O-alkylated nucleosides used as building blocks of antisense oligonucleotides for subsequent potential applications in therapeutics (antisense oligonucleotide synthesis) and diagnostics. The phosphodiesterase from the Gram-negative enteric bacterium S. marcescens was selected on account of the broad substrate range and high activity of the enzyme. The protein was purified by heat-treatment of the crude cell-free extract, followed by column chromatography (gel filtration). It was characterised and showed optimal activity at a broad pH range (pH 6.8-9.4, with a peak at ca. pH 8.5) and at a temperature of 60-65 degrees C. No metal ions were required for activity, although Ba2+ was an activator. Conversion of 2'-O-methoxyethyl cAMP into the corresponding nucleoside derivative on a multi-gram scale was successfully performed in two steps, using the S. marcescens enzyme in conjunction with a commercially available alkaline phosphatase from Escherichia coli. PMID- 15349701 TI - Improvement of pCOR plasmid copy number for pharmaceutical applications. AB - Production of pharmaceutical-grade plasmid DNA is becoming important as the demand for clinical batches is steadily growing. pCOR plasmids have been specifically designed and used for gene delivery into humans, and have been produced by high cell-density fermentation with a yield of 100 mg/l. This yield could probably be increased as long as the release specifications of bulk plasmid remain the same, particularly in terms of plasmid sequence. We report here the use of genetic approaches in Escherichia coli to increase the copy number of pCOR. The bacterial gene encoding the pi initiator-protein, which plays a pivotal role in pCOR replication, was mutagenized. A fluorescence-based screening methodology in E. coli was used to identify novel copy-up mutations. A particular combination of copy-up mutations translated into a 3-5-fold increase in monomer pCOR plasmid DNA per biomass unit. PMID- 15349702 TI - Patterns of gene divergence and VL promoter activity in immunoglobulin light chain clusters of the channel catfish. AB - The structure and genomic organization of V, J, and C segments in multiple gene clusters of the G class of catfish light (L) chain were determined to study evolutionary patterns of cluster divergence and regulatory function. The results showed that the organizational pattern is conserved; two VL segments reside in opposite transcriptional orientation to a pseudogene JL segment (psiJ), a functional JL segment, and a CL segment. Structures within the central V-psiJ-J-C regions indicate that cluster duplication occurred after the V-V-psiJ-J-C organization became fixed. VL divergence in gene clusters subsequently occurred by mechanisms that principally targeted complementarity-determining regions. The sequence of the VL-flanking regions, which contained regulatory octamer, TATA, and Pax-5 (BSAP) binding site motifs, was conserved during G cluster duplication and within VL-flanking regions in divergent lineages of bony fish. Reporter assays of catfish B cells transfected with a 742-bp VL-flanking fragment showed promoter activity in the absence of enhancer elements. The promoter's activity doubled when coupled with the catfish IgH enhancer. A 136-bp fragment containing the motifs conserved in bony fish phylogeny and located between the leader initiation codon and the initiation sites of sterile transcripts served as a minimal promoter and provided the highest B-cell activity. These constructs, however, did not act as promoters in catfish non-lymphoid cells or mammalian BJA B B cells or fibroblasts. These results indicate that the structure and function of VL promoter regions in the regulation and tissue specificity of L-chain gene expression evolved early in phylogeny. PMID- 15349703 TI - Enhancement to the RANKPEP resource for the prediction of peptide binding to MHC molecules using profiles. AB - We introduced previously an on-line resource, RANKPEP that uses position specific scoring matrices (PSSMs) or profiles for the prediction of peptide-MHC class I (MHCI) binding as a basis for CD8 T-cell epitope identification. Here, using PSSMs that are structurally consistent with the binding mode of MHC class II (MHCII) ligands, we have extended RANKPEP to prediction of peptide-MHCII binding and anticipation of CD4 T-cell epitopes. Currently, 88 and 50 different MHCI and MHCII molecules, respectively, can be targeted for peptide binding predictions in RANKPEP. Because appropriate processing of antigenic peptides must occur prior to major histocompatibility complex (MHC) binding, cleavage site prediction methods are important adjuncts for T-cell epitope discovery. Given that the C-terminus of most MHCI-restricted epitopes results from proteasomal cleavage, we have modeled the cleavage site from known MHCI-restricted epitopes using statistical language models. The RANKPEP server now determines whether the C-terminus of any predicted MHCI ligand may result from such proteasomal cleavage. Also implemented is a variability masking function. This feature focuses prediction on conserved rather than highly variable protein segments encoded by infectious genomes, thereby offering identification of invariant T-cell epitopes to thwart mutation as an immune evasion mechanism. PMID- 15349704 TI - Alternative splicing of major histocompatibility complex class II DXB transcripts in Xiphophorus fishes. AB - Classical MHC class II glycoproteins present peptides to T cells. In Xiphophorus fishes and in the guppy, Poecilia reticulata, a classical MHC class II B-like transcript has been identified, DAB, as well as a divergent MHC class II B-like transcript, DXB. In the two species of Xiphophorus fishes studied here, X. multilineatus and X. pygmaeus, alternative splicing of the DXB transcript was observed, but not of the classical type DAB transcripts. Two alternative splice patterns were found: a 16-codon deletion and a five-nucleotide deletion that leads to an extension of the transcript. A single DXB transcript that terminates before the transmembrane region was also observed. The alternative splice pattern and the divergence of DXB from DAB suggest that in fish, DXB may have an alternate function. Alternative splicing transcripts of DXB may allow for signaling and localization of DXB within the cell. PMID- 15349705 TI - Effects of dosage and CYP2D6-mutated allele on plasma concentration of paroxetine. AB - OBJECTIVE: We investigated the effect of dosages of paroxetine and cytochrome P450 (CYP) 2D6 genotypes on the plasma concentration of paroxetine in Japanese patients being treated with paroxetine. METHODS: Blood samples were collected from 73 individuals after at least 2 weeks of the same daily dose of paroxetine. The plasma paroxetine concentration was measured using HPLC, and the CYP2D6 genotypes were identified by PCR. Genotype groups were compared by one-way analysis of variance at different paroxetine doses. RESULTS: The mean plasma paroxetine concentrations at daily doses of 10, 20, 30, and 40 ng/ml were 6.6+/ 7.4, 34.9+/-26.8, 74.8+/-37.2, and 130.5+/-96.8 ng/ml, respectively, showing a disproportionate and nonlinear increase in plasma drug levels of paroxetine upon increasing doses. Plasma paroxetine concentrations in patients with CYP2D6*10 alleles were significantly higher than those without *10 allele at 10 mg/day (7.3+/-6.11 vs. 2.99+/-3.52 ng/ml), but there was no significant difference between *1/ *1, *1/ *10 and *10/ *10 genotypes at the higher doses. Similarly, patients with CYP2D6*5 alleles showed higher plasma paroxetine concentrations than those without *5 allele, although differences in the plasma paroxetine concentration did not reach statistical significance level because of the small number of subjects with *5 alleles. CONCLUSIONS: Our results indicate the possibility of saturation in paroxetine metabolism with an increase in paroxetine dose, and that CYP2D6*10 allele(s) have significant impact on plasma paroxetine concentration at low doses in Japanese population. PMID- 15349706 TI - Fatal intoxication cases: cytochrome P450 2D6 and 2C19 genotype distributions. AB - OBJECTIVE: Many commonly used pharmaceuticals, such as antidepressants and neuroleptics as well as some illegal drugs, are metabolised by the cytochrome P450 enzyme debrisoquine 4-hydroxylase (CYP2D6). Of Caucasians, 7-10% lack this enzyme, which can, upon administration of drugs in normal therapeutic doses, lead to adverse reactions and unexpected intoxication, leading in turn even to a fatal outcome in some cases. METHODS: Individuals (n=242) who had died due to intoxication by pharmaceuticals were genotyped for CYP2D6 and CYP2C19 and compared with a reference group of 281 blood donors. A single nucleotide polymorphism (SNP) method was used to identify five CYP2D6 alleles: *1 (wt), *2, *3, *4 and *6. The allele *5, a complete gene deletion, was identified by a multiplex amplification of long DNA fragments. Four CYP2C19 alleles *1 (wt), *2, *3 and *4 were also identified by SNP analysis. RESULTS: The prevalence of the CYP2D6 poor metaboliser (PM) genotypes in individuals with fatal intoxication was lower (4.7%) than expected from the frequencies of these genotypes in the blood donors (8.5%). A significantly lower frequency P<0.005 (0.03 with correction according to Bonferroni) was found for the CYP2D6*4 allele among the fatal intoxication cases. The CYP2C19 genotype analyses showed the same results for the fatal intoxication cases and for the blood donors. CONCLUSIONS: The findings in this study confirm our earlier observations of a lower frequency of CYP2D6 PM genotypes in cases of fatal intoxication. To our knowledge, it has not been shown previously that intoxication victims might have a lower frequency of PMs than the general population. PMID- 15349707 TI - Esomeprazole 40 mg provides more effective intragastric acid control than lansoprazole 30 mg, omeprazole 20 mg, pantoprazole 40 mg and rabeprazole 20 mg in patients with gastro-oesophageal reflux symptoms. AB - OBJECTIVE: To compare the effect of esomeprazole 40 mg with lansoprazole 30 mg, omeprazole 20 mg, pantoprazole 40 mg and rabeprazole 20 mg on intragastric pH during single and repeated dosing in four separate studies in patients with symptoms of gastro-oesophageal reflux disorder (GERD). METHODS: In four randomised crossover studies, patients with symptoms of GERD received once-daily treatment with esomeprazole 40 mg or lansoprazole 30 mg (study A), omeprazole 20 mg (study B), pantoprazole 40 mg (study C) and rabeprazole 20 mg (study D) for 5 days. Continuous 24-h intragastric pH recording was performed on days 1 (except study B) and 5. Percentage of time over 24 h with intragastric pH greater than 4, 24-h median pH and the proportion of patients with pH greater than 4 for greater than or equal to 12 h and 16 h during the 24-h recording periods were investigated. RESULTS: In all four studies, esomeprazole 40 mg OD maintained intragastric pH greater than 4 for a significantly higher mean percentage of the 24-h period compared with all other proton pump inhibitors (PPIs) on days 1 (esomeprazole 40.6% versus lansoprazole 33.4%, P=0.0182; esomeprazole 50.3% versus pantoprazole 29.1%, P<0.001; esomeprazole 41.0% versus rabeprazole 29.4%, P=0.002) and 5 (esomeprazole 57.7% versus lansoprazole 44.5%, P<0.0001; esomeprazole 69.8% versus omeprazole 43.7%, P<0.0001; esomeprazole 67.0% versus pantoprazole 44.8%, P<0.001; esomeprazole 59.4% versus rabeprazole 44.5%, P<0.0001). Higher 24-h median pH and a higher proportion of patients with intragastric pH greater than 4 for greater than or equal to 12 h and 16 h were reported with esomeprazole 40 mg OD than with all the other PPIs in each study. CONCLUSION: Esomeprazole 40 mg provides greater acid control in more patients and maintains intragastric pH greater than 4 for a longer period than lansoprazole 30 mg, omeprazole 20 mg, pantoprazole 40 mg and rabeprazole 20 mg in patients with symptoms of GERD. PMID- 15349708 TI - A new opportunity for thalidomide? Further randomised controlled trial are necessary. PMID- 15349709 TI - The three-dimensional vestibulo-ocular reflex evoked by high-acceleration rotations in the squirrel monkey. AB - The aim of this study was to determine if the angular vestibulo-ocular reflex (VOR) in response to pitch, roll, left anterior-right posterior (LARP), and right anterior-left posterior (RALP) head rotations exhibited the same linear and nonlinear characteristics as those found in the horizontal VOR. Three-dimensional eye movements were recorded with the scleral search coil technique. The VOR in response to rotations in five planes (horizontal, vertical, torsional, LARP, and RALP) was studied in three squirrel monkeys. The latency of the VOR evoked by steps of acceleration in darkness (3,000 degrees /s(2) reaching a velocity of 150 degrees /s) was 5.8+/-1.7 ms and was the same in response to head rotations in all five planes of rotation. The gain of the reflex during the acceleration was 36.7+/-15.4% greater than that measured at the plateau of head velocity. Polynomial fits to the trajectory of the response show that eye velocity is proportional to the cube of head velocity in all five planes of rotation. For sinusoidal rotations of 0.5-15 Hz with a peak velocity of 20 degrees /s, the VOR gain did not change with frequency (0.74+/-0.06, 0.74+/-0.07, 0.37+/-0.05, 0.69+/ 0.06, and 0.64+/-0.06, for yaw, pitch, roll, LARP, and RALP respectively). The VOR gain increased with head velocity for sinusoidal rotations at frequencies > or =4 Hz. For rotational frequencies > or =4 Hz, we show that the vertical, torsional, LARP, and RALP VORs have the same linear and nonlinear characteristics as the horizontal VOR. In addition, we show that the gain, phase and axis of eye rotation during LARP and RALP head rotations can be predicted once the pitch and roll responses are characterized. PMID- 15349710 TI - Analytical characterization of bioactive fluoropolymer ultra-thin coatings modified by copper nanoparticles. AB - Copper-fluoropolymer (Cu-CFx) nano-composite films are deposited by dual ion-beam sputtering. The extensive analytical characterization of these layers reveals that inorganic nanoparticles composed of Cu(II) species are evenly dispersed in a branched fluoropolymer matrix. In particular, X-ray photoelectron spectroscopy has been employed to study the surface chemical composition of the material and to assess how it changes on increasing the copper loading in the composite. Transmission electron microscopy reveals that the copper nanoclusters have a mean diameter of 2-3 nm and are homogeneously in-plane distributed in the composite films. Electrothermal atomic absorption spectroscopy has been used to study the kinetics of copper release in the solutions employed for the biological tests. The Cu-CFx layers are employed as bioactive coatings capable of inhibiting the growth of target microorganisms such as Saccharomyces cerevisiae, Escherichia coli, Staphylococcus aureus, and Lysteria. The results of the analytical characterization enable a strict correlation to be established among the chemical composition of the material surface, the concentration of copper dissolved in the microorganisms broths, and the bioactivity of the nano-structured layer. PMID- 15349711 TI - Selection of useful predictors in multivariate calibration. AB - Ten techniques used for selection of useful predictors in multivariate calibration and in other cases of multivariate regression are described and discussed in terms of their performance (ability to detect useless predictors, predictive power, number of retained predictors) with real and artificial data. The techniques studied include classical stepwise ordinary least-squares (SOLS), techniques based on the genetic algorithms, and a family of methods based on partial least-squares (PLS) regression and on the optimization of the predictive ability. A short introduction presents the evaluation strategies, a description of the quantities used to evaluate the regression model, and the criteria used to define the complexity of PLS models. The selection techniques can be divided into conservative techniques that try to retain all the informative, useful predictors, and parsimonious techniques, whose objective is to select a minimum but sufficient number of useful predictors. Some combined techniques, in which a conservative technique is used to perform a preliminary selection before the use of parsimonious techniques, are also presented. Among the conservative techniques, the Westad-Martens uncertainty test (MUT) used in Unscrambler, and uninformative variables elimination (UVE), developed by Massart et al., seem the most efficient techniques. The old SOLS can be improved to become the most efficient parsimonious technique, by means of the use of plots of the F statistics value of the entered predictors and comparison with parallel results obtained with a data matrix with random data. This procedure indicates correctly how many predictors can be accepted and substantially reduces the possibility of overfitting. A possible alternative to SOLS is iterative predictors weighting (IPW) that automatically selects a minimum set of informative predictors. The use of an external evaluation set, with objects never used in the elimination of predictors, or of "complete validation" is suggested to avoid overestimate of the prediction ability. PMID- 15349712 TI - Chemometrics in analytical chemistry. PMID- 15349713 TI - Universal life defense concept: global biology. PMID- 15349714 TI - Nutrient-specific effects in the coordination of cell growth with cell division in continuous cultures of Saccharomyces cerevisiae. AB - Cell cycle progression of Saccharomyces cerevisiae cells was monitored in continuous cultures limited for glucose or nitrogen. The G1 cell cycle phase, before initiation of DNA replication, did not exclusively expand when growth rate decreased. Especially during nitrogen limitation, non-G1 phases expanded almost as much as G1. In addition, cell size remained constant as a function of growth rate. These results contrast with current views that growth requirements are met before initiation of DNA replication, and suggest that distinct nutrient limitations differentially impinge on cell cycle progression. PMID- 15349715 TI - Tetrahydrofolate-specific enzymes in Methanosarcina barkeri and growth dependence of this methanogenic archaeon on folic acid or p-aminobenzoic acid. AB - Methanogenic archaea are generally thought to use tetrahydromethanopterin or tetrahydrosarcinapterin (H4SPT) rather than tetrahydrofolate (H4F) as a pterin C1 carrier. However, the genome sequence of Methanosarcina species recently revealed a cluster of genes, purN, folD, glyA and metF, that are predicted to encode for H4F-specific enzymes. We show here for folD and glyA from M. barkeri that this prediction is correct: FolD (bifunctional N5,N10-methylene-H4F dehydrogenase/N5,N10-methenyl-H4F cyclohydrolase) and GlyA (serine:H4F hydroxymethyltransferase) were heterologously overproduced in Escherichia coli, purified and found to be specific for methylene-H4F and H4F, respectively (apparent Km below 5 microM). Western blot analyses and enzyme activity measurements revealed that both enzymes were synthesized in M. barkeri. The results thus indicate that M. barkeri should contain H4F, which was supported by the finding that growth of M. barkeri was dependent on folic acid and that the vitamin could be substituted by p-aminobenzoic acid, a biosynthetic precursor of H4F. From the p-aminobenzoic acid requirement, an intracellular H4F concentration of approximately 5 M was estimated. Evidence is presented that the p-aminobenzoic acid taken up by the growing cells was not required for the biosynthesis of H4SPT, which was found to be present in the cells at a concentration above 3 mM. The presence of both H4SPT and H4F in M. barkeri is in agreement with earlier isotope labeling studies indicating that there are two separate C1 pools in these methanogens. PMID- 15349716 TI - Complexity of phenotypes and symbiotic behaviour of Rhizobium leguminosarum biovar trifolii exopolysaccharide mutants. AB - Rhizobium leguminosarum biovar trifolii strain TA1 polysaccharide synthesis (pss) mutants in the pssD, pssP, pssT and pssO genes and altered in exopolysaccharide (EPS) synthesis were investigated. EPS-deficient mutants were also changed in lipopolysaccharide structure. All mutants exhibited varied sensitivities to detergents, ethanol and antibiotics, thus indicating changes in bacterial membrane integrity. Using pss mutants marked with the gusA gene, EPS-deficient mutants were found to have abnormalities in nodule development and to provoke severe plant defence reactions. The pss mutants that produced altered quantities of EPS with a changed degree of polymerisation generally occupied the younger developmental zones of the nodules and elicited moderate plant defence reactions. PMID- 15349717 TI - The impact of thiopurine S-methyltransferase polymorphisms on azathioprine dose 1 year after renal transplantation. AB - Azathioprine metabolism is influenced by activity of the enzyme thiopurine S methyltransferase (TPMT), which varies markedly between individuals. In this study we examined the influence of TPMT gene polymorphisms on azathioprine dose 1 year after renal transplantation. TPMT coding and promoter genotypes were determined using PCR-based assays. Azathioprine dose, white cell count, and intercurrent events throughout the first year after renal transplantation were ascertained from contemporaneous clinical notes. All patients analysed ( n=172) received an initial azathioprine dose of 1.5 mg/kg per day. Twelve individuals with one variant TPMT coding allele were detected (*3A n=11, *3C n=1). Of these, 58% required azathioprine dose reduction because of leucopenia, compared to only 30% of homozygous wild-type patients ( P=0.04). A significant correlation between the presence of >/=11 variable number tandem repeats (VNTRs) in the TPMT promoter and reduction in azathioprine dose was also identified ( P=0.001). We concluded that when azathioprine is administered at an initial dose of 1.5 mg/kg per day, both coding and promoter TPMT polymorphisms influence the dose tolerated. PMID- 15349718 TI - Molecular and clinical response to angiotensin II receptor antagonist in kidney transplant patients with chronic allograft nephropathy. AB - Chronic allograft nephropathy (CAN) represents an important cause of graft loss after kidney transplantation. TGF-beta1 is a key factor in fibrogenesis, and the angiotensin II receptor antagonist losartan may decrease the intra-graft synthesis of TGF-beta1. The aim of this study was to determine the clinical and molecular effect of losartan in kidney transplant patients (KTPs) with CAN. We studied nine KTPs, after the first year of transplantation, with proteinuria (more than 500 mg/24 h), stable renal function, and histological signs of CAN. Immunosuppression was cyclosporine, azathioprine, and corticoids. Kidney biopsy was performed in all patients at the beginning of the study and 12 weeks after treatment with 50 mg/day of losartan. Quantitation of intra-graft expression of TGF-beta1 was performed in all biopsies, by real-time PCR. After losartan treatment there were no differences in patients' BP and blood creatinine level. The proteinuria significantly dropped to 414.2+/-377 mg/24 h, P=0.001. Intra graft expression of TGF-beta1 was decreased after treatment. In conclusion, losartan significantly decreases the intra-graft expression of TGF-beta1 and proteinuria in KTPs with CAN. PMID- 15349719 TI - Lymphadenectomy prior to rat hind limb allotransplantation prevents graft-versus host disease in chimeric hosts. AB - In previous rat studies, the use of mixed allogeneic chimerism (MAC) to induce host tolerance to hind limb allografts has resulted in severe graft-versus-host disease (GVHD). The purpose of this study was to determine if immunocompetent cells in bone marrow (BM) and/or lymph nodes (LNs) of transplanted limbs were responsible for inducing GVHD in mixed chimeric hosts. [ACI-->Wistar Furth] chimeric rats received ACI hind limbs that were non-irradiated, irradiated (1050 cGy) or lymphadenectomized. Rejection, GVHD and donor chimerism was assessed. Chimeric hosts rejected none of their limbs. However, hosts of non-irradiated hind limbs succumbed to GVHD 22.4+/-0.8 days after transplantation. In contrast, chimeras that received irradiated or lymphadenectomized ACI hind limbs showed no clinical or histological signs of GVHD at 5 months. We conclude that mixed chimeric hosts are susceptible to GVHD due to the immunocompetent cell load provided by the LNs, not the BM, of hind limb allografts. PMID- 15349720 TI - Blockade of 4-1BB (CD137)/4-1BB ligand interactions increases allograft survival. AB - We investigated the role of 4-1BB, a T cell co-stimulatory molecule, in alloimmune responses. In vivo mixed lymphocyte reactions showed that 4-1BB was preferentially expressed on actively dividing CD4(+) and CD8(+) T cells. Furthermore, following alloantigen challenge, the draining lymph nodes contained subpopulations of 4-1BB-expressing CD4(+) and CD8(+) T cells. 4-1BB-deficient C57BL/6 mice showed a delayed rejection of cardiac transplants mismatched for the major histocompatibility complex. Longer transplant survival was induced by blockade of 4-1BB/4-1BB ligand (4-1BBL) interactions using an anti-4-1BBL monoclonal antibody. Histological analysis showed that prolonged transplant survival in the 4-1BB-deficient and anti-4-1BBL-treated mice correlated with reduced lymphocytic infiltration and vasculitis in the donor heart tissue. Taken together, our data suggest that blockade of 4-1BB/4-1BBL interactions inhibited the expansion of alloreactive T cells and reduced CTL activity against host alloantigen, which in turn resulted in the prolongation of allograft survival. Blockade of the 4-1BB co-stimulatory pathway may be useful for preventing allograft rejection. PMID- 15349721 TI - Prevalence of anal HPV infection in solid-organ transplant patients prior to immunosuppression. AB - Patients that undergo organ transplantation have a high risk of developing various malignancies, depending on the duration and magnitude of immunosuppressive therapy. Among others, a 10-fold increased relative risk has been reported for the development of anal cancer. There is a strong association between persistent infection with high-risk mucosal types of human papillomavirus (HPV) and anogenital neoplasia. In this study we analysed the prevalence of anal HPV infection in organ transplant patients before starting immunosuppressive therapy. In a university transplant unit, patients ( n=60, 40 male, 20 female) that were undergoing solid-organ transplantation (kidney, liver) for the first time were routinely screened for anal HPV infection. Anal swabs were obtained within 24 h after transplantation and analysed for the presence of mucosal-type HPV DNA by liquid DNA/RNA hybridization [hybrid capture (HC) 2 test]. Overall, some type of HPV DNA was detected in 14/60 (23.3%) patients; 9/60 (15%) were positive for high-risk HPV and 8/60 (13.4%) were positive for low-risk HPV, and 3/60 (5%) were positive for both types. Prevalence of HPV infection tended to be higher in patients that were receiving liver transplants than in those receiving kidney transplants (29.4% vs. 20.9%), but the difference did not reach statistical significance. In our series of organ transplant patients the prevalence of previous HPV infection (23.3%) before immunosuppressive therapy was started was higher than that found in previous epidemiological studies or in a control group. In particular, there was a high rate (15%) of infection with oncogenic HPV types. These findings have important implications on screening and surveillance policies in this patient group at risk of developing neoplasias, including anal cancer. PMID- 15349722 TI - Intragraft iNOS induction during human liver allograft rejection depresses cytochrome p450 activity. AB - Allograft function may become impaired during rejection after human liver transplantation. Cytokines induce nitric oxide (NO) production in hepatocytes, Kupffer cells and infiltrating mononuclear cells. NO inhibits cytoplasmatic cytochrome p450 (CYP) enzyme activity in vitro. It is not known whether this mechanism plays a role in vivo. In order to characterize the role of locally produced cytokines in the pathogenesis of liver dysfunction, we analysed human liver transplant biopsy material for the expression of proinflammatory cytokines as well as for NO synthase and we compared these results to the microsomal liver function in vivo [aminopyrine breath test (ABT)] and in vitro (enzymatic analysis of CYP). Microsomal liver function decreased in vivo during rejection while ABT levels decreased by 40% and increased again by 59% after the acute rejection episode. Similarly, CYP 1A2 and 2E1 activity dropped 42% and 24% in rejecting samples, respectively. Competitive reverse transcriptase polymerase chain reaction (RT-PCR) showed a fivefold upregulation of interferon gamma (IFN-gamma) gene expression. Inducible, but not constitutive NO-synthase gene expression was upregulated fivefold in samples from rejecting patients suggesting a local induction of NO in response to immune events. Our data show a marked impairment of CYP enzyme activity during allograft rejection which is presumably secondary to an increased intragraft production of proinflammatory cytokines and NO. PMID- 15349723 TI - Hepatic abscesses after liver transplantation secondary to traumatic intrahepatic bile duct injuries in a cadaveric allograft donor. AB - We report the case of an ultimately successful liver transplant recipient whose post-transplant course was complicated by the early development of multiple abscesses in the graft. Post-transplant cholangiography identified multiple shear injuries to the second and third order intrahepatic bile ducts, originating from blunt trauma to the donor liver. Treatment was non-operative following recent reports of the successful management of intrahepatic bile duct injury in the stable trauma patient. This discussion adds to the limited literature available on the transplantation of injured donor livers, despite this being a relatively common practice. Further experience is needed in determining the appropriate criteria for the use of traumatized donor livers. Cholangiography carried out on the back table may help to determine if such injured livers are suitable for transplantation. PMID- 15349724 TI - Septic diaphragmatic dysfunction is prevented by Mn(III)porphyrin therapy and inducible nitric oxide synthase inhibition. AB - OBJECTIVE: Decreased diaphragmatic contractility and organ failure observed during sepsis is mediated by an overproduction of nitric oxide ((.)NO)-derived species, mitochondria being a major target of oxidative and nitrative stress. We tested the potential protective effects of (a) a novel synthetic antioxidant, the manganese(III) 5,10,15,20-tetrakis(N-ethylpyridinium-2-yl) porphyrin (MnTE-2 PyP(5+)) and (b) the inducible (.)NO synthase inhibitor aminoguanidine (AG) on a rat model of sepsis. SETTING: University research laboratories. SUBJECTS AND INTERVENTIONS: Sepsis was induced by cecal ligation and perforation in rats. MEASUREMENTS AND RESULTS: Systemic hemodynamics, pulmonary gas exchange, in vitro diaphragmatic function and mitochondrial respiration were evaluated. Moreover, plasma and mitochondrial oxidative and nitrative stress parameters were investigated. Sepsis determined diaphragmatic dysfunction and a significant decrease in mitochondrial coupling and respiration. Oxidative stress was evidenced by decreased plasma antioxidants and increased lipid oxidation. Tyrosine nitration was increased in the plasma and mitochondria of the septic animals. These alterations were ameliorated or prevented by either MnTE-2-PyP(5+) or AG. CONCLUSIONS: Our results demonstrate that overproduction of (.)NO and (.)NO-derived reactive species play a critical role in mitochondrial impairment and diaphragmatic function during sepsis. More importantly, AG but mainly the novel metalloporphyrin MnTE-2-PyP(5+) were able to ameliorate diaphragmatic and mitochondrial dysfunction and could contribute to preventing organ failure during severe sepsis. PMID- 15349725 TI - Expression of uncoupling protein-3 mRNA in rat skeletal muscle is acutely stimulated by thiazolidinediones: an exercise-like effect? AB - AIMS/HYPOTHESIS: We examined whether thiazolidinediones (TZDs) acutely affect uncoupling protein-3 ( UCP-3) expression in skeletal muscle and plasma NEFA in Sprague-Dawley rats. METHODS: Expression of UCP-3 mRNA in hindlimb muscles and plasma NEFA were measured after a single intraperitoneal injection of TZDs in healthy male rats. RESULTS: Independent of which TZD was injected (50 micromol/kg), UCP-3 expression in gastrocnemius muscle was distinctly increased after 6 h (increase vs vehicle-injected control: pioglitazone, 10.3+/-3.2-fold, p=0.03; rosiglitazone, 8.7+/-1.2-fold, p=0.001; RWJ241947, 9.5+/-2.7-fold, p=0.03). This was accompanied by elevated plasma NEFA (control 158+/-13 micromol/l; pioglitazone, 281+/-40 micromol/l, p=0.03; rosiglitazone, 276+/-27 micromol/l, p=0.005; RWJ241947, 398+/-51 micromol/l, p=0.004). The increase in plasma NEFA could in part have mediated TZD-induced UCP-3 expression, but increased UCP-3 mRNA was also found in isolated muscle after 2 h of TZD exposure in vitro (25 micromol/l pioglitazone, 1.7+/-0.3-fold, p=0.046), suggesting that TZDs act directly and independently of NEFA on skeletal muscle. CONCLUSIONS/INTERPRETATION: In healthy rats, a single dose of TZDs rapidly increases UCP-3 mRNA in skeletal muscle and plasma NEFA. This effect resembles the acute response to a bout of exercise. PMID- 15349726 TI - Metabolic adaptations to chronic glucose infusion in rats. AB - AIMS/HYPOTHESIS: Several studies have employed the chronic glucose infusion protocol to quantify the metabolic adaptations associated with a prolonged glucose challenge. However, the limited number of indices and time points reported by these studies has generated an incomplete picture of this process. In this study we aimed to generate an integrative and dynamic picture of the physiological adaptations that occur during chronic glucose infusion. METHODS: Sprague-Dawley rats were infused with either 50% dextrose or saline (2 ml/h) for a period of between 0 and 6 days. Glucose, insulin and NEFA dynamics were determined from daily blood samples. Subsets of animals were killed daily for histological determination of beta cell mass, size and replication rates. The mathematical model of coupled beta cell mass, insulin and glucose (the betaIG model) was used to estimate insulin sensitivity, beta cell function and net neogenesis from this data. RESULTS: Glucose-infused rats displayed transient hyperglycaemia, persistent hyperinsulinaemia and unchanged NEFA levels. Insulin sensitivity decreased by approximately 80% during the first day of glucose infusion, but had returned to pre-infusion levels by Day 3. Beta cell function was four to six times higher than in control rats throughout the experiment. Beta cell mass doubled over the 6 days of glucose infusion due to three phases of adaptation: (i) neogenesis; (ii) hypertrophy and hyperplasia; and (iii) continued hyperplasia coupled to a second wave of neogenesis. CONCLUSIONS/INTERPRETATION: Contrary to the results reported for perfused pancreas and in vitro experiments, we found that chronic glucose infusion elevated beta cell function. The prediction of a second wave of beta cell neogenesis, coupled with our previous report of "focal areas" on Day 3, suggests the existence of delayed acinar-to islet transdifferentiation. PMID- 15349727 TI - Association between the A-2518G polymorphism in the monocyte chemoattractant protein-1 gene and insulin resistance and Type 2 diabetes mellitus. AB - AIMS/HYPOTHESIS: The molecular mechanisms of obesity-related insulin resistance are incompletely understood. Macrophages accumulate in adipose tissue of obese individuals. In obesity, monocyte chemoattractant protein-1 (MCP-1), a key chemokine in the process of macrophage accumulation, is overexpressed in adipose tissue. MCP-1 is an insulin-responsive gene that continues to respond to exogenous insulin in insulin-resistant adipocytes and mice. MCP-1 decreases insulin-stimulated glucose uptake into adipocytes. The A-2518G polymorphism in the distal regulatory region of MCP-1 may regulate gene expression. The aim of this study was to investigate the impact of this gene polymorphism on insulin resistance. METHODS: We genotyped the Ludwigshafen Risk and Cardiovascular Health (LURIC) cohort ( n=3307). Insulin resistance, estimated by homeostasis model assessment, and Type 2 diabetes were diagnosed in 803 and 635 patients respectively. RESULTS: Univariate analysis revealed that plasma MCP-1 levels were significantly and positively correlated with WHR ( p=0.011), insulin resistance ( p=0.0097) and diabetes ( p<0.0001). Presence of the MCP-1 G-2518 allele was associated with decreased plasma MCP-1 ( p=0.017), a decreased prevalence of insulin resistance (odds ratio [OR]=0.82, 95% CI: 0.70-0.97, p=0.021) and a decreased prevalence of diabetes (OR=0.80, 95% CI: 0.67-0.96, p=0.014). In multivariate analysis, the G allele retained statistical significance as a negative predictor of insulin resistance (OR=0.78, 95% CI: 0.65-0.93, p=0.0060) and diabetes (OR=0.80, 95% CI: 0.66-0.96, p=0.018). CONCLUSIONS/INTERPRETATION: In a large cohort of Caucasians, the MCP-1 G-2518 gene variant was significantly and negatively correlated with plasma MCP-1 levels and the prevalence of insulin resistance and Type 2 diabetes. These results add to recent evidence supporting a role for MCP-1 in pathologies associated with hyperinsulinaemia. PMID- 15349728 TI - What can the HLA transgenic mouse tell us about autoimmune diabetes? AB - Type 1 diabetes mellitus is a polygenic disease strongly associated with the class II molecules DR3, 4 and the linked DQ2, 8 alleles. These molecules play an important role in presentation of peptide antigens after intracellular processing to CD4 T lymphocytes. A number of in vitro approaches have been used to elucidate the molecular basis for the association of particular HLA alleles with susceptibility to or protection from Type 1 diabetes mellitus. These have focused on the structure of the antigen-presenting molecules, together with their peptides. Binding studies, peptide elution, molecular modelling and crystallisation of the peptide MHC complex have between them made it possible to define the peptide-binding regions and to examine the stability of binding of peptides from putative autoantigens. It is difficult to study the role of these molecules in vivo in humans, and HLA transgenic mice have been generated to overcome this problem. Studies of mice expressing the HLA class II alleles associated with diabetes have shown that the presence of HLA molecules alone does not cause disease except in the presence of an islet "insult", even when this "insult" would in itself be insufficient to precipitate disease in the absence of the HLA class II transgene. HLA transgenic mice offer a way to elucidate the in vivo role of these molecules, and could help the development of targeted immunotherapy. PMID- 15349729 TI - [Whole-body tumor staging: MRI or FDG-PET/CT?]. AB - Technical innovations such as the availability of movable patient platforms and the introduction of 32-channel systems have made whole-body MRI for tumor staging feasible. In addition, the development of dual-modality PET/CT systems offers the opportunity to perform anatometabolic tumor staging with whole-body coverage in a single session. Based on an increase in patient survival when applying a stage adapted therapy in different malignant diseases the relevant question relates to the accuracies of whole-body MRI and whole-body PET/CT for TNM staging. This review article addresses whole-body tumor staging with MRI and FDG-PET/CT with special emphasis on diagnostic accuracies for staging different malignant diseases. PMID- 15349730 TI - [Possibilities of whole-body MRI for investigating musculoskeletal diseases]. AB - This contribution outlines possibilities and limitations of whole-body MRI for investigating musculoskeletal diseases. Benefits and drawbacks of the novel whole body MRI technology are discussed and a possible whole-body MRI sequence protocol for musculoskeletal examinations is proposed. Muscle, joint and bone diseases are discussed in which the application of whole-body MRI may be of advantage. Particularly, polymyositis, muscledystrophy, rheumatoid arthritis, spondylitis ancylosans, multiple trauma, skeletal metastases, multiple myeloma and malignant lymphoma are mentioned. Whole-body MRI opens new advantages for the examination of multifocal musculoskeletal diseases. The clinical benefit of this method for particular diseases has to be evaluated in further studies, however. PMID- 15349731 TI - [Cardiovascular whole-body MRI: possibilities and limitations in prevention]. AB - Cardiovascular disease is a major challenge to the healthcare with increasing prevalence in western societies. Hence, early detection of cardiovascular pathologies and preventative strategies will experience growing relevance in the future. Magnetic resonance imaging (MRI) nowadays allows a comprehensive analysis of the cardiovascular system. By combining separate examinations of brain, arterial vasculature, and heart the technique permits early detection of pathological changes with high diagnostic accuracy void of adverse events. Such a protocol has been proven feasible and technically robust and can be performed within 45 min. Inherent limitations are low spatial resolution of whole-body MR angiography and lack of functional stress testing of the heart. However, while being suitable as a fast and comprehensive imaging technique for cardiovascular screening purposes, medical consequences and socioeconomic relevance must further be elucidated. PMID- 15349732 TI - [Comparison of high resolution whole-body MRI using parallel imaging and PET-CT. First experiences with a 32-channel MRI system]. AB - PURPOSE: To compare the accuracy in the detection and staging of various malignant tumors with high resolution whole-body MRI using parallel imaging with whole-body dual-modality PET-CT. PATIENTS AND METHODS: Preliminary results of an interim analysis from a prospective, blinded study are presented, in which 20 patients (mean age 59 years, range 27-77 years) with different oncological diseases underwent whole-body dual modality FDG-PET-CT screening for tumor search or staging in case of confirmed or suspected metastatic disease. All patients also underwent whole-body MRI imaging with the use of parallel imaging (iPAT). High-resolution coronal T1w- and STIR-sequences of 5 body levels with 512 x 512 matrix, axial fast T2w imaging of lung and abdomen (HASTE), contrast-enhanced dynamic and static T1w-sequences of liver, brain, abdomen, and pelvis were performed. Using a 32-channel whole-body MRI scanner (Magnetom Avanto, Siemens Medical Solutions) with a total field of view of 205 cm and free table movement, all patients could be covered from head to toe within one examination. With this technique, high spatial resolution and acceptable scanning times could be obtained. Two experienced radiologists read the MRI-scans, one radiologist and one nuclear scientist read PET-CT scans, each in consensus in a clinical setting. Delineation of the primary tumor (T-stage) or recurrent tumor, pathologic lymph node involvement, as well as degree and localization of metastatic disease, was assessed using PET-CT as standard of reference. RESULTS: Metastases from gastrointestinal tumor (25%) and breast cancer (25%), genitourinary tumor (15%) and malignant melanoma (15%) were detected. In 4/20 patients the primary tumor was identified, 2/20 patients showed recurrent tumor. Of 140 malignant lesions detected by PET-CT, 124 lesions were detected with MRI, resulting in a sensitivity of 89% at a specificity of 86%. In malignant lymph node detection, sensitivity of MRI was 83% and specificity 85%. CONCLUSION: Whole-body MRI is a promising technique in the detection of primary tumor and metastatic disease. Sensitivity in the assessment of lymph node metastases seems to be limited. With the use of parallel imaging (iPAT), dedicated high-resolution whole-body MRI is possible within acceptable scanning times. PMID- 15349733 TI - [Oncologic screening with whole-body MRI: possibilities and limitations]. AB - In the last decade the interest in radiological screening examination increased among informed laymen enormously. Independent from the evidence of whole-body examinations for cancer prevention the discussion about screening must again be considered again due to the newest technical developments, since MRI of the whole body with high spatial resolution is feasible now within one single examination. The newest system permits simultaneous connection of up to 76 coil elements and signal reception from 32 independent receiving channels. Whole-body MRI including magnetic resonance colonography (MRC) is feasible within 60 min. In this review potential investigation protocols will be presented. Potentials, challenges and limitations of whole-body MRI in the prevention of the malignancies most frequently leading to death are discussed on the basis own experiences examples and the literature. PMID- 15349734 TI - [The "Lector"--a testing method to determine verbal educational level]. AB - Assessment of the educational level is important to prevent false neuropsychological diagnostic decisions. In the German-speaking countries some instruments for this assessment already exist, but there are problems involved in the practical application. There still is no reading test, which is a common standard in international neuropsychological test batteries. The lector investigates how subjects read rare German words or foreign words used in German speaking countries. Words were selected which are pronounced differently from German pronunciation rules. A subject who does not know the word will pronounce it false, i.e., according to the German pronunciation rule. Norms (percentiles), estimation of reliability, and first data about validity were calculated from three samples with a total of 503 subjects. On the basis of the data presented, the Lector Test can be used as a control variable in research with groups as well as in neuropsychological diagnostics. PMID- 15349735 TI - [On the value of neuropsychological short tests in epileptology]. AB - Cognitive deficits are common in patients with epilepsy refractive to therapy and have considerable influence on the quality of life in this group of patients. Therefore, neuropsychological investigations should play an important role in the comprehensive evaluation of patients with chronic epilepsy. The aim of this study was to examine whether a reliable screening for cognitive deficits in these patients may be reduced to the assessment of two bedside tests. In a prospective study we analyzed the results of 40 patients with epilepsy refractive to therapy subjected to a 45-min neuropsychological screening battery and compared them with the results of a short battery consisting of two bedside tests. Using the screening battery as the gold standard, the short battery had a sensitivity of 50% and a specificity of 100%. Changing the criteria for pathological results in the short battery, sensitivity could be raised to 81.25% but specificity fell to 50%. Therefore, bedside tests instead of longer neuropsychological testing cannot be recommended as a screening method for cognitive deficits in patients with chronic epilepsy. PMID- 15349736 TI - [Magnetic resonance spectroscopy of brain tumours]. AB - Magnetic resonance spectroscopy facilitates non-invasive determination of metabolic changes in vivo. The main metabolites are the neuronal marker N acetylaspartate (NAA), cholines reflecting membrane turnover, creatine, lactate, and mobile lipids. Primary brain tumours exhibit reduced NAA and increased choline resonances compared to normal brain, and these abnormalities increase with higher malignancy. Increasing choline resonances on follow-up studies correlate with tumour progression, whereas the reduction of initially increased choline resonances indicates a transition from viable tumour to necrotic tissue. Metastases as non-neuroectodermal tumours lack NAA, but demonstrate elevated choline, lactate and lipid resonances. Lymphomas are characterised by massively increased lipid resonances with markedly elevated choline. Prominent alanine resonances are often observed in meningioma. Cystic/necrotic lesions demonstrate elevated lactate regardless of their aetiology. The characteristic finding of prominent resonances from acetate, succinate, and alanine, of leucine, isoleucine and valine in untreated bacterial abscesses allows the differentiation of bacterial abscesses from cystic/necrotic brain tumours. PMID- 15349737 TI - [Severe burn as a consequence of an epileptic seizure while showering]. AB - We report a case of a 17-year-old patient who experienced an epileptic seizure while showering. The patient suffered full-thickness scald injuries involving thorax, abdomen and both arms and legs. The burn, which included 35% of the patient's body surface area, will cause life-long secondary functional and aesthetic deformities. By installing adjustable shower equipment with a maximal temperature limit, this devastating accident could have been prevented. PMID- 15349738 TI - A decrease in iron status in young healthy women after long-term daily consumption of the recommended intake of fibre-rich wheat bread. AB - BACKGROUND: Fibrerich bread and cereals are included in the recommendations of a healthy diet. OBJECTIVE: To measure the effects of long-term consumption of the recommended intake of fibre-rich wheat bread on the iron status of young healthy women with adequate iron stores. DESIGN: Four-months intervention study including healthy female subjects assigned into two groups provided daily with 300 g of fibre-rich wheat bread, prepared with or without phytase as a supplement to their habitual diet. SUBJECTS: Forty-one women aged 24.8 +/- 3.8 years (mean +/- SD) and an average BMI of 22.0 +/- 2.9 kg/m2 participated. Baseline values for serum ferritin were 45 microg/L, 22-83 (geometric mean, range) and for haemoglobin 132 g/L, 119-148 (arithmetic mean, range), respectively. RESULTS: Distribution of energy intake from protein, fat and carbohydrate, and daily intake of dietary fibre and iron were similar in the two groups and within the recommended levels. There was no effect of the phytase added to the wheat bread on the iron status of the subjects, but an effect of the intervention period. Serum ferritin and haemoglobin levels were significantly reduced by 12 +/- 1.1 microg/L (27%) (P < 0.001) and 2 +/- 0.8 g/l (1.5%) (mean +/- SE) (P < 0.05) respectively, after four months of intervention. CONCLUSIONS: The present long-term study indicates that consumption of the recommended daily intake of fibre-rich wheat bread results in an impairment of iron status in women with initially sufficient iron stores. Reduction of the phytic acid concentration in the bread was not sufficient to maintain iron status. PMID- 15349739 TI - Metachronous colorectal cancer: necessity of post-operative colonoscopic surveillance. AB - BACKGROUND AND AIMS: The aim of this study was to identify the occurrence and analyze the characteristics of metachronous colorectal cancers, and to compare the characteristics of these cases (index tumor) with the control group to find any predicting factor that may influence the occurrence of metachronous cancer. PATIENTS AND METHODS: The database of colorectal cancer in the Veterans General Hospital-Taipei, from January 1981 to September 2001 was reviewed. In total, 3,846 cases of adenocarcinoma of the colon and rectum, which received curative resection during this period, were found. The criteria of metachronous cancer were: occurrence more than 12 months after curative surgery; with pre-operative complete colonoscopy or one negative post-operative colonoscopic follow-up to rule out synchronous tumor; tumor arising from mucosa at a site other than anastomosis. The age, gender of the patients, the location, pathological characteristics of the metachronous tumors, occurrence of associated adenomas, the number of lesions, and the tumor stage were analyzed and compared with the control group. RESULTS: In total, 43 cases of metachronous cancer were identified, giving an annual incidence of 0.18%. The distribution of the location of the index tumor of metachronous cases was predominantly left-sided, which was not different from that of the control group. The mean duration of occurrence of metachronous cancer after the primary operation was 71+/-46.6 months. The association of adenomas had no relationship with the occurrence of the metachronous cancer. No significant predicting factors for the development of metachronous tumors were found. CONCLUSION: Lifelong regular post-operative colonoscopic surveillance is essential for colorectal cancer patients. PMID- 15349740 TI - Rectovaginal fistula after double-stapled transanal rectotomy (STARR) for obstructed defaecation. PMID- 15349741 TI - Late-presenting congenital diaphragmatic hernia in children: a clinical spectrum. AB - Articles dealing with late-presenting congenital diaphragmatic hernia (CDH) in children published until 2003 were collectively reviewed. The main purpose of the presented study was to gain insight into the clinical spectrum of this variant of CDH and to discuss its clinical and surgical aspects. In 125 articles, 362 patients were reported. Left posterolateral hernia was the dominant anatomical type, found in 79.4% of the children. Male-to-female distribution was nearly 2:1 in both right- and left-sided CDH. Associated anomalies were noted in 8.6% of children. Sixty-five percent of children with right CDH presented within the 1st year of life, whereas only 41.7% of patients with left CDH belonged to this age group. The children with right CDH presented with chronic symptoms in 57.4% of cases, while among those with left CDH, acute presentation prevailed, constituting 60.5% of this group. No strict correlation between age and clinical symptoms was noted, although the youngest patients tended to present with respiratory symptoms. Late-presenting diaphragmatic hernia is associated with a wide range of clinical symptoms occurring in various constellations. Dyspnea and vomiting were the most frequent symptoms found in patients with either chronic or acute manifestation. Preoperative mortality was 3.9% (14 children). In 45 children (12.4%), complications resulting from diaphragmatic hernia were noted, and in a further 11 children iatrogenic preoperative complications were noted. Among 152 children with reported CDH, true hernia with sac was found in 32.7%. Eight children (2.2%) died in the early postoperative period, while a further 13 patients (3.6%) had postoperative complications requiring surgical management. Recurrence of hernia was noted in seven children (1.9%). This collective review of basic epidemiological and clinical data from a large group of patients seems to support a hypothesis for a congenital nature of the diaphragmatic defect and an acquired nature of "herniation" of the viscera. The wide clinical spectrum of late-presenting CDH seems to be conditioned by two factors: timing of herniation and type of intraabdominal viscera displaced into the chest. PMID- 15349742 TI - Huge arachnoid cyst in the posterior fossa: controversial discussion for selection of the surgical approach. AB - CASE REPORT: We report a case of a huge posterior fossa arachnoid cyst with hydrocephalus. This 15-month-old girl presented with macrocrania and psychomotor retardation. Magnetic resonance (MR) imaging demonstrated ventriculomegaly and a huge cystic lesion in the posterior fossa. A membranectomy was performed microscopically and the patient had a good postoperative course. OUTCOME: Magnetic resonance imaging 1 year after the operation revealed adequate reduction in the size of the ventricle and cyst. Progressive macrocrania discontinued and normal development was obtained. PMID- 15349743 TI - Surgical evacuation of acute subdural hematoma improves cerebral hemodynamics in children: a transcranial Doppler evaluation. AB - OBJECTIVE: The objective was to evaluate cerebral hemodynamics in young children with acute subdural hematoma (SDH) and the impact of surgical treatment using transcranial Doppler (TCD). DESIGN: The design was a prospective study of infants with SDH requiring surgical evacuation. SETTING: The setting was the neuro intensive care unit of a university hospital. INTERVENTIONS: Indications for surgical evacuation were based upon clinical and radiological arguments. Surgery included emergency needle aspiration followed by external or/and internal shunting as required. A TCD evaluation was performed before needle aspiration, and after each surgical drainage procedure. It included a pressure provocation test to assess cerebral compliance. Preoperative and postoperative middle cerebral artery (MCA) velocities, Gosling pulsatility (PI) and Pourcelot resistivity (RI) indexes and compliance were compared with Student's t-test, or Fisher's exact test as indicated. MEASUREMENTS AND MAIN RESULTS: Out of 26 infants, 23 (88%) had injuries that had possibly been inflicted, and 3 had accidental injuries. Initial TCD evaluation demonstrated intracranial hypertension with decreased diastolic velocity, increased PI and RI, and decreased compliance. Surgical evacuation resulted in statistically significant improvement in cerebral hemodynamics (diastolic velocity: 17.2+/-10 cm/s vs. 31.1+/-10 cm/s, p<0.0015, PI: 2.5+/-1.3 vs. 1.4+/-0.8, p<0.002, RI: 0.8+/-0.2 vs. 0.6+/-0.1, p<0.005) in all but 3 infants, who eventually died. Surgical drainage (primary shunting or external drainage) was needed in 23 infants and resulted in further improvement in cerebral hemodynamics. Finally, 73% of the infants made a good recovery. CONCLUSIONS: Children with acute bilateral HSD have a high incidence of increased intracranial pressure as assessed by TCD. Surgical evacuation improves cerebral hemodynamics. TCD could be used for assessing the need for, and the efficiency of surgical drainage. PMID- 15349744 TI - Brain stem abscesses in childhood. AB - BACKGROUND: Solitary brain stem abscess is a rare condition with high mortality and morbidity. These lesions were considered to be invariably fatal before 1974 when advanced diagnostic tools were not available. Recently, the diagnosis and prognosis of brain stem abscesses have been modified by the introduction of modern radiological tools, and several patients with a favorable outcome have been reported since then. Because the pons is the most common site of the abscesses, involvement of the sixth and seventh nerves is frequent. The midbrain is the second most likely location, and medullary abscesses are distinctly rare. Treatment of a brain stem abscess includes medical therapy alone, open microsurgical intervention, or stereotactic aspiration. CASE REPORT: We report a case of a 7-year-old girl with a solitary brain stem abscess. Her neurological examination revealed involvement of the cranial nerves and pyramidal tracts. Microsurgical exposure and aspiration of the abscess resulted in rapid improvement in her neurological condition and radiological resolution of the lesion. We discuss this uncommon case to draw attention to the importance of early diagnosis and adequate treatment, and we review the relevant literature. PMID- 15349745 TI - Responses of neurons in the electrosensory lateral line lobe of the weakly electric fish Gnathonemus petersii to simple and complex electrosensory stimuli. AB - Mormyrid fish use active electrolocation to detect and analyze objects. The electrosensory lateral line lobe in the brain receives input from electroreceptors and an efference copy of the command to discharge the electric organ. In curarized fish, we recorded extracellularly from neurons of the electrosensory lateral line lobe while stimulating in the periphery with either a local point stimulus or with a more natural whole-body stimulus. Two classes of neurons were found: (1) three types of E-cells, which were excited by a point stimulus; and (2) two types of I-cells, which were inhibited by point stimulus and responded with excitation to the electric organ corollary discharge. While all neurons responded to a point stimulus, only one out of two types of I-units and two of the three types of E-units changed their firing behavior to a whole body stimulus or when an object was present. In most units, the responses to whole-body stimuli and to point stimuli differed substantially. Many electrosensory lateral line lobe units showed neural plasticity after prolonged sensory stimulation. However, plastic effects during whole body stimulation were often unlike those occurring during point stimuli, suggesting that under natural conditions electrosensory lateral line lobe network effects play an important role in shaping neural plasticity. PMID- 15349746 TI - Long-distance, low-frequency elephant communication. AB - The production, transmission, and reception of and the behavioral response to long-distance, low-frequency sound by elephants is reviewed. The structure of low frequency calls generated by elephants is separated into the "source" and the "filter" roles played by the lungs, larynx and vocal track, the composition of the expired air and the ambient air temperature. Implications regarding the size, age, sex, sexual and physical status follow from the call structure and detection. Reception of the signal is discussed in terms of the characteristics of the elephant's ear with particular attention to the determination of the threshold of hearing and the ability to locate the source of low-frequency sounds. Factors which influence the transmission of near infrasound are related to atmospheric structure. The critical role played by the thermal stratification and vertical gradient and magnitude of the wind in determining both the range and the detection of a signal are discussed for open and closed elephant habitats. Infrasound plays a pervasive role in reproduction, resource utilization, avoidance of predation and other social interactions. Current and future technology can be expected to contribute to the detection and interpretation of elephant communication. This will aid in the understanding of behavior and in efforts to sustain the species. PMID- 15349747 TI - [Importance of family examination in juvenile X-linked retinoschisis]. AB - BACKGROUND: Congenital (juvenile) retinoschisis belongs to the group of hereditary vitreoretinopathies. This disorder is inherited in an X-linked recessive pattern and its onset usually occurs in 5- to 10-year-old boys. Presenting clinical signs include decreased visual acuity due to maculopathy. CASE REPORT: The authors present a case of a 17-year-old boy with decreased visual acuity, hypermetropia, and bilateral retinoschisis with maculopathy upon fundus examination. In view of a 50% risk of the disorder occurring in the brothers of the affected male, they underwent full ophthalmological and electrophysiological examinations (until then asymptomatic). In one of them decreased visual acuity, mixed astigmatism, and maculopathy were present, without any changes of the peripheral retina. In the youngest brother decreased visual acuity, hypermetropia, and maculopathy were diagnosed. CONCLUSIONS: Genetic counseling and ophthalmological examination of family members at risk facilitated early recognition of the pathological changes in the siblings. Genetic counseling with pedigree analysis and genetic analysis, if possible, should be offered to all affected patients and family members. PMID- 15349748 TI - Urolithiasis in enterocystoplasties. AB - The incidence of stones in patients with enterocystoplasty is reported as 12 52.5%. Most patients will have multiple physical factors such as immobility, need for self catheterisation and poor urine drainage, so that it is not certain that an intestinal reservoir is the cause of stones on its own. There is little or no evidence that mucus is an aetiological factor. Foreign bodies in the reservoir, such as staples, increase the risk of stone formation from 13% to 43%. Stones require surgical removal. Minimally invasive techniques may be used for small stones. A low velocity disintegrator is required so that fragments are not propelled into the intestinal mucosa. Stones are infective in origin in 86% of cases, but 14% are sterile. Metabolic screen shows that 80% of patients have risk factors for at least three different types of stone. All patients have raised pH (mean 6.93) and hypocitraturia. Raised serum and urinary calcium, hyperoxaluria and hyperuricosuria are found in up to 33% of patients. PMID- 15349749 TI - Single oligonucleotide nested PCR: a rapid method for the isolation of genes and their flanking regions from expressed sequence tags. AB - We report on the development of a new PCR technique for the isolation of genomic fragments that flank known DNA sequences. This technique, single oligonucleotide nested (SON)-PCR, relies on only two amplification reactions with two or three nested sequence-specific primers. It allows the isolation of DNA regions located on either side of a known DNA sequence, with high specificity. DNA products of 2 kb in size can be generated that all contain one copy of the same primer at both ends. Sequence analysis of these products indicates that the binding of the primers to non-specific DNA sites mainly depends on their overall complementarity to the target sequence. Moreover, analysis shows that short extensions of the primers can occur during the first amplification reaction and that a 2-bp overlap between subsequent primers can target their annealing to their predecessor's sequence. Ninety percent of the DNA products larger than 0.5 kb correspond to fragments of interest and we obtained successful results with various templates and primer sets. SON-PCR therefore seems a very efficient and widely applicable method for the rapid identification of large unknown DNA regions. Based on available expressed sequence tags, this technique was applied to isolate the palH and pacC genes of the phytopathogenic fungus Botrytis cinerea, with their 5' or 3' flanking regions. PMID- 15349750 TI - Multiple group I introns detected in the nuclear small subunit rDNA of the autosporic green alga Selenastrum capricornutum. AB - A phylogenetic investigation of the autosporic chlorophycean alga species Selenastrum capricornutum using the small subunit (SSU) rRNA gene revealed the unusual presence of six group IC1 introns. Previous studies showed that numerous green algal taxa contain group IC1 introns. However, whereas some algal species harbor multiple introns in a single ribosomal gene, none have contained as many as S. capricornutum. Three of the S. capricornutum introns are located at conserved algal intron sites and the remaining three are located at novel eukaryotic positions. The SSU rRNA genes and their introns have been sequenced and putative secondary structures are proposed for the introns. Also, their similarity to other group IC1 introns from algal, fungal, and viral sources is investigated. Results suggest the initial presence of introns at conserved locations, followed by duplication and insertion to novel positions within the SSU rRNA gene. PMID- 15349751 TI - [Comparison of chromosomal defects in primary tumor and metastases by comparative genomic hybridization (CGH)]. AB - Molecular methods can complement the classical methods in pathology like macroscopic, histological and immunohistochemical examinations.Comparative genomic hybridization is a cytogenetic method to screen for gains and losses of chromosomal material in tumor cells. This method allows defect studies of archival paraffin-embedded and formalin-fixed tumor material. CGH can detect gains and losses of chromosomal material that are at least 10 to 20 megabases in size. This genome-wide screening method allows to study the cytogenetic relationship between differently located tumors of a patient. To answer the question if these different tumors are metastases of the primary lung tumor or independent primary tumors CGH analysis is a supplementary method that introduces new prospectives. PMID- 15349752 TI - A 150-kDa glycoprotein isolated from Solanum nigrum L. has cytotoxic and apoptotic effects by inhibiting the effects of protein kinase C alpha, nuclear factor-kappa B and inducible nitric oxide in HCT-116 cells. AB - This study was carried out to investigate the anticancer effects of a 150-kDa glycoprotein isolated from Solanum nigrum L. (SNL glycoprotein) on spontaneously and experimentally induced tumor promotion in HCT-116 cells. For spontaneously induced tumor promotion, we evaluated the cytotoxic and apoptotic effects in HCT 116 cells using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT), DNA fragmentation, and H33342 and ethidium bromide staining assays. SNL glycoprotein had remarkable, dose-dependent cytotoxic and apoptosis-inducing effects at low concentrations. For experimentally induced tumor promotion, we investigated whether the SNL glycoprotein was able to regulate the activity of protein kinase C alpha (PKCalpha), the DNA binding activation of nuclear factor kappa B (NF-kappaB), the activity of NF-kappaB protein, and the production of nitric oxide (NO) in HCT-116 cells stimulated with 12-O-tetradecanoylphorbol 13 acetate (TPA) using electrophoretic mobility shift assays (EMSA), Western blot analysis, and NO assays. As expected, SNL glycoprotein dose-dependently inhibited PKCalpha translocation, NF-kappaB DNA binding activity, NF-kappaB protein activity and NO production in HCT-116 cells stimulated with TPA (61.68 ng/ml, 100 nM). Collectively, these results suggest that SNL glycoprotein can induce apoptosis through the modulation of signal mediators. Therefore, we speculate that it could be used as a chemotherapy agent even at low concentrations in HCT 116 cells. PMID- 15349753 TI - A phase I trial of gemcitabine plus cladribine in patients with advanced hematologic malignant diseases. AB - PURPOSE: Gemcitabine and cladribine (2CdA) are nucleoside analogues that decrease DNA synthesis via inhibition of ribonucleotide reductase; the combination could be additive or synergistic. We conducted a dose escalation study to establish the maximum tolerable doses (MTD) of gemcitabine and 2CdA when given in combination in patients with advanced hematologic malignancies and to describe the toxicity profile of this combination. PATIENTS AND METHODS: A total of 45 patients with advanced hematologic diseases were enrolled into two groups. Group A had adequate baseline hematopoiesis, defined as absolute neutrophil count (ANC) >1 x 10(9)/l and platelet count >50 x 10(9)/l. Group B did not meet these criteria. Hematologic dose-limiting toxicity (DLT) for group A was defined as grade 4 neutropenia or thrombocytopenia lasting >28 days; group B was not evaluated for hematologic toxicity. Nonhematologic DLT was defined similarly for both groups. Death occurring during the first cycle of treatment was considered a DLT event only if it was related to drug toxicity. Gemcitabine was administered as a 4-h intravenous infusion once every 28 days. 2CdA was administered over 1 h daily for the first 3 days of each 28-day cycle. RESULTS: The MTD was not reached in either group. Myelosuppression was common, but not dose-limiting. Febrile neutropenia and infections were also common, particularly in group B, and judged in most cases to be due to bone marrow failure at baseline. Nonhematologic toxicities were generally mild, and skin rash, the most frequently observed, was dose limiting in one patient enrolled in each group. Four deaths (three during the first cycle of treatment) occurred at the highest dose level tested in group B (gemcitabine 5000 mg/m2 and 2CdA 16 mg/m2). Although only one of these deaths was dose-limiting by stated criteria, this dose level did not appear to be safely tolerated in this patient population. Several responses were observed in patients with Hodgkin's disease. CONCLUSIONS: The combination of gemcitabine and 2CdA is feasible in patients with hematologic malignancies. Phase II studies of this combination should be considered, particularly in patients with Hodgkin's disease. PMID- 15349754 TI - Parameters for predicting allogeneic PBSCT outcome of acute myeloid leukemia: cytogenetics at presentation versus disease status at transplantation. AB - In addition to the clinical disease status at transplantation, the cytogenetic risk at presentation also provides critical information for predicting the prognosis or deciding the future therapeutic strategy. As such, the current study examined various parameters, including the cytogenetics at presentation and clinical disease status at transplantation, regarding their effect on the transplant outcomes of acute myeloid leukemia (AML) patients in an allogeneic peripheral blood stem cell transplantation (PBSCT) setting. A total of 36 patients receiving an allogeneic PBSCT from matched sibling donors were included in a state of first complete remission (CR) (n=22, 61%) or beyond the first CR (n=14, 39%). The cytogenetic risk was classified according to Medical Research Council (MRC) 10 criteria: favorable, 7 patients (20%); intermediate, 21 patients (58%); unfavorable eight patients (22%). The 3-year overall survival rates were 80% for the favorable, 63% for the intermediate, and 0% for the unfavorable cytogenetic risk groups (p=0.0002), and 62% for the patients in a state of first CR and 35% for those beyond the first CR (p=0.0524). Multivariate analysis revealed that higher CD34+ cell doses, favorable cytogenetics at presentation, and a lower marrow blast percentage at transplantation were all strongly associated with favorable transplant outcomes, including overall survival (OS), progression-free survival (PFS), and the probability of progression. The cytogenetic risk at presentation was found to be a useful parameter in predicting the transplant outcomes for patients with AML, regardless of the clinical disease status. However, an additive innovative therapeutic strategy is still needed to overcome an unfavorable cytogenetic risk with refractory AML after allogeneic PBSCT. PMID- 15349755 TI - Computed tomographic demonstrated infarcts after surgical and endovascular treatment of aneurysmal subarachnoid hemorrhage. AB - BACKGROUND: Cerebral infarction is a sequela of vasospasm. Other etiologies for infarction after subarachnoid hemorrhage (SAH), however, have not been well studied. To determine the incidence and etiologies for infarction after SAH, we reviewed the head CT scans of all SAH patients at our center from 1993-2000. METHODS: From 1993-2000, 679 consecutive patients were admitted with SAH, of which 619 patients underwent surgical or endovascular treatment. Two reviewers examined the head CT scans of all 619 patients for new infarct. Clinical outcome was collected from a prospective database. FINDINGS: 505 patients were treated with surgical clipping; 114 with endovascular coiling. There were CT findings of new infarct in 189 patients (30%): 140 in the surgical group (28%) and 49 in the endovascular group (43%). The etiologies for infarct in the surgical group were vasospasm 79 (15%), perforator occlusion 40 (8%), large vessel occlusion 14 (3%), elevated intracranial pressure 4 (1%), thromboembolism 2 (0.4%), and systemic hypotension 1 (0.2%). Infarcts in the endovascular group were due to vasospasm 20 (18%), thromboembolism 12 (11%), large vessel occlusion/dissection 9 (8%), elevated intracranial pressure 4 (4%), perforator occlusion 3 (3%), and systemic hypotension 1 (1%). Hunt Hess Grade (P < 0.001), Fisher Score (P < 0.0001), and MGH Grade (P < 0.001) were significantly associated with CT-demonstrated infarct. There was no significant difference in incidence of CT-infarcts when the period 1993-1996 was compared to 1997-2000. CONCLUSIONS: Despite advances in the treatment of SAH, there is still a significant incidence of associated radiographic infarcts. Hunt Hess Grade, Fisher Score, and MGH Grade were significantly associated with CT-demonstrated infarct. PMID- 15349756 TI - Intradural metastatic spinal cord compression from ACTH-secreting pituitary carcinoma. AB - Pituitary carcinomas are uncommon and intradural metastatic spread to the spine is rarer still. We describe a 27-year old man with metastatic spinal cord compression from an adrenocorticotrophic hormone (ACTH) cell pituitary carcinoma, 16 years following the initial presentation. He had three previous resections of the pituitary tumour and post-operative radiotherapy. The intradural, extramedullary spinal metastases causing thoracic and lumbar cord compression were excised, with neurological improvement. Spinal metastases in pituitary carcinoma are uncommon, but aggressive surgical resection of the spinal metastases produces good symptomatic relief. PMID- 15349757 TI - Surgical occlusion of the leptomeningeal drainage is a safe and effective treatment for intracranial dural arteriovenous fistulas without dural sinus drainage. PMID- 15349758 TI - Stereotactically guided microsurgical removal of colloid cysts. AB - BACKGROUND: The surgical technique and clinical results for a series of 16 consecutive patients who underwent resection of third ventricular colloid cysts through a stereotactically guided cylindrical retractor are presented. METHODS: Between March 1993 and December 2002, 16 patients, 11 males and 5 females with a mean age of 36, were admitted with colloid cysts of the third ventricle. Four patients had undergone previous surgery, of which two were simple aspirations, one endoscopic aspiration, and one transcallosal partial removal. Four patients required emergency ventriculostomies on admission. In all patients the foramen of Monro was targeted using a Leksell stereotactic frame. A coronal craniotomy three to four cm in diameter was performed and a cylindrical retractor 14 mm in diameter was advanced to the target. Microsurgical removal of the cyst was then performed through the retractor. FINDINGS: Total removal of the cyst was achieved in all cases. Median follow-up time is 42 months. Complete resolution of symptoms occured in all patients. Control magnetic resonance imaging revealed no residual cysts. All patients have returned to their previous occupations without sequelae or epilepsy. CONCLUSION: The outcome obtained in this series has shown that transfrontal transforaminal total removal of colloid cysts through a stereotactically guided cylindrical retractor is a safe procedure. PMID- 15349759 TI - Type III odontoid fracture with pseudarthrosis in a skeleton from the early Middle Ages. PMID- 15349760 TI - Osteolytic intra-osseous meningiomas: illustrated review. AB - While bone invasion and hyperostosis are frequent phenomena in meningiomas, primary intra-osseous meningiomas are rare. With only 15 reported cases, the osteolytic form of primary intra-osseous meningiomas is most uncommon. Its occurrence in the skull base is an extra-ordinary exception. We have reviewed and categorized the pertinent literature on intra-osseus meningiomas with special emphasis on osteolytic tumours and discuss their clinical implications on the basis of a new case located in the petrous bone without contact with the meninges. It is concluded that due to their different clinical, radiological and pathological features, hyperostotic and osteolytic variants of intra-osseus meningiomas should be distinguished from tumours with soft tissue components and from en-plaque lesions. PMID- 15349761 TI - [The new IHS classification. Background and structure]. AB - The diagnosis of headache is nowadays based on the classification criteria of the International Headache Society (IHS). This classification can be seen in a historical context with a very first version published by an ad-hoc-committee of the National Institute of Neurological Diseases and Blindness in the USA. In 1988 the first IHS version appeared, and this was revised in its current version in 2003. The classification is divided into primary (i.e., idiopathic) and secondary (i.e., symptomatic) headache and into cranial neuralgias and facial pain. The classification is based on the description of the headache features and thus a phenomenological rather than an etiological classification. Furthermore, not the patient as a subject but the headache as a clinical entity is classified. Therefore, some patients can have more than one headache diagnosis. In total, 14 different headache groups and more than 170 headache types are separated. The changes compared to the last classification (such as introduction of chronic migraine and of several new rare idiopathic headache disorders) and the perspectives for future classifications (e.g., problems of genetic classification) are discussed. PMID- 15349762 TI - Urinary excretion of calcium and phosphate in preterm infants. AB - The aims of this study were to determine reference ranges for the urinary calcium (UCa/Cr) and phosphate (UPO(4)/Cr) creatinine ratios and to study factors influencing these ratios in a representative population of preterm infants managed according to current nutritional guidelines. Spot urine samples were obtained from 186 preterm infants (gestation 24-34 weeks) for measurement of UCa/Cr and UPO(4)/Cr ratios as part of a routine metabolic bone screening program, once every 2-4 weeks from the 3rd to the 18th week of life. Data were also collected on gender, appropriate or small for gestational age (SGA), nutrition [total parenteral nutrition (TPN), preterm or term formula, and breast milk], plasma Ca, P0(4), urea, and electrolytes and on the use of drugs (frusemide, dexamethasone, and theophylline). Data from infants treated with any of these three drugs were analyzed separately and not included in establishing the reference ranges for UCa/Cr and UPO(4)/Cr. The mean gestational age of the study population was 28 weeks (range 24-34 weeks). The 95th percentile for UCa/Cr at 3 weeks of age was 3.8 mmol/mmol and decreased significantly with increasing postnatal age (P<0.001). The 95th per-centile for UPO(4)/Cr was 26.69 mmol/mmol at 3 weeks of age, but this did not change significantly with increasing postnatal age (P=0.296). On univariate analysis there was no significant association of UCa/Cr and UPO(4)/Cr with gender and type of enteral nutrition. The UCa/Cr was lower in infants who were SGA (P=0.013) and with low plasma Ca (P=0.008). Infants on TPN had significantly higher UCa/Cr (P =0.019) and lower UPO(4)/Cr ratios(P<0.001). Multivariate analysis confirmed the decrease in UCa/Cr ratio with increasing postnatal age, but the SGA effect was eliminated. The use of furosemide(P<0.001) and theophylline (P=0.003) was associated with a significant increase in the UCa/Cr ratio. The use of dexamethasone was also associated with an increase in UCa/Cr ratio, but this did not achieve statistical significance (P=0.339). The use of furosemide, theophylline,and dexamethasone had no effect on UPO(4)/Cr. We report a reference range for UCa/Cr and UPO(4)/Cr ratios and factors influencing these ratios in a representative population of preterm infants between 24 and 34 weeks gestation, managed according to current nutritional guide-lines. PMID- 15349763 TI - Optimising nutrition in chronic renal insufficiency--progression of disease. AB - There is a lack of evidence to support the belief that dietary measures are beneficial in slowing the progression of chronic renal insufficiency (CRI). We prospectively monitored nutrient intakes and progression of CRI over a 2-year period in children aged 2-16 years with differing levels of severity of CRI, as part of their ongoing joint medical/dietetic care. Children were grouped following [5'Cr]-labelled EDTA glomerular filtration rate(GFR, ml/min per 1.73 m 2) estimations, into 'normal'kidney function [GFR >75, mean 106 (SD 19.5), n=58],providing baseline data only, mild (GFR 51-75, n=25),moderate (GFR 25-50, n =21), and severe (GFR <25, n=19) CRI. Children with CRI were followed for 2 years,with 51 completing the study (19 mild, 19 moderate, 13 severe CRI) and were excluded if they subsequently required dialysis. Regular medical and dietary advice was provided and yearly 3-day semi-quantitative dietary di-aries and baseline and 6-monthly measurements of blood pressure and urinary protein/creatinine ratio were obtained. Mean reductions in estimated GFR over 2 years were -9.4, -5.8, and -6.0 ml/min per 1.73 m2 for mild,moderate, and severe CRI, respectively. Mean systolic blood pressure standard deviation score (SDS) fell significantly in all groups by 0.7 SDS, whereas there was little change in proteinuria. From reported dietary intakes,median sodium intakes increased (+10 mmol/day) and protein intakes decreased (-0.4 g/kg per day). Median phosphate intakes did not change significantly, where as calcium intakes fell in all groups, with an overall median of -20% reference nutrient intake (RNI) (F=33.3,P<0.001). Of children with moderate CRI, 65% finished with calcium intakes below 80% RNI, and parathyroid hormone (PTH) concentrations significantly increased in this group (F=6.0, P=0.021). Higher phosphate and sodium intakes were associated with greater deterioration in estimated GFR in children with mild CRI (r2=0.30,P=0.02; r-=0.31, P=0.02, respectively). There was no such correlation for protein intake or PTH. This study emphasises the need for a joint medical and dietetic approach and indicates a number of interventions other than protein restriction, which could be commenced early in children with CRI in an attempt to delay progression. PMID- 15349764 TI - Reference values for urinary calcium and phosphorus to prevent osteopenia of prematurity. AB - The prevention of osteopenia of prematurity is an important issue in the care of preterm infants. Fetal bone mineral accretion has been achieved in preterm infants by establishing and maintaining a simultaneous slight excretion of calcium (Ca) and phosphorus (P) (urine concentrations of 1-2 mmol/l) by means of an individual supplementation with Ca and/or P, resulting in a slight surplus supply (SSS). In this issue, Aladangady et al. present associations between urinary Ca/Cr and PO(4)/Cr ratios of preterm infants and biochemical variables of bone mineral metabolism. However, to date it has not been proven that these variables are a reliable substitute for direct measurement of bone mineral content (BMC). Before Ca/Cr and PO(4)/Cr ratios can be recommended as a new reference for improving BMC, the following steps are required: (1) direct measurement of BMC, (2) a prospective interventional trial to test and compare this new reference with the existing one (SSS, urinary Ca and P of 1-2 mmol/l) investigating BMC as primary outcome, and (3) adequate proof that Ca and P/Cr ratios are superior to simple urinary Ca and P concentrations. PMID- 15349765 TI - A novel mutation of WT1 exon 9 in a patient with Denys-Drash syndrome and pyloric stenosis. AB - We report a novel mutation in WT1 exon 9 (1214 A>G) resulting in an amino acid change from H to R at codon 405 in a 46 XY female patient who had congenital hypertrophic pyloric stenosis, pseudohermaphroditism masculinus, renal failure, and Wilms tumor, and died at the age of 22 months. The patient demonstrated the difficulty in diagnosing a patient with intersex before conclusive genetic characterization. PMID- 15349766 TI - Differential seed and seedling predation by crabs: impacts on tropical coastal forest composition. AB - Recently, the importance of seed predation by crabs on mangrove species distributions and densities has been established by several studies. In a tropical coastal terrestrial forest in Costa Rica, we investigated the relative importance of predation by land crabs, Gecarcinus quadratus, and hermit crabs, Coenobita compressus, on measured forest composition through a series of seed removal and seedling establishment experiments. We also used natural light-gaps and adjacent non-gap sites to test how canopy cover affects crab predation (seed removal) and seedling establishment. We found fewer tree species (S=18) and lower densities (seedlings, saplings, and adults) in the coastal zone within 100 m of coastline, than in the inland zone (S=59). Land crab densities were higher in the coastal zone (3.03+/-1.44 crabs m(-2)) than in the inland zone (0.76+/-0.78 crabs m(-2)), and hermit crabs were not present in the inland zone. Seed removal and seedling mortality also were higher in the coastal zone than in the inland zone, and in the open controls than in the crab exclosures. Mortality of seeds and seedlings was two to six times higher in the controls than exclosures for four of the five experiments. Crabs preferred seeds and younger seedlings over older seedlings but showed no species preferences in the seed (Anacardium excelsum, Enterolobium cyclocarpum, and Terminalia oblonga) and seedling (Pachira quinata and E. cyclocarpum) stages. We conclude that the observed differences in tree densities were caused by differential crab predation pressure along the coastal gradient, while the differences in species composition were due to predator escape (satiation) by seed quantity. Canopy cover did not affect seed removal rates, but did affect seedling survival with higher mortality in the non-gap versus gap environments. In summary, crab predation of seeds and seedlings, and secondarily canopy cover, are important factors affecting tree establishment in terrestrial coastal forests. PMID- 15349767 TI - Ontogenic cell death in the nigrostriatal system. AB - Like most neural systems, dopamine neurons of the substantia nigra undergo apoptotic natural cell death during development. In rodents, this occurs largely postnatally and is biphasic with an initial major peak just after birth and a second minor peak on postnatal day 14. As envisioned by classic neurotrophic theory, this event is regulated by interactions with the target of these neurons, the striatum, because a developmental target lesion results in an augmented natural cell death event with fewer nigral dopamine neurons surviving into adulthood. Until recently, the striatal target-derived neurotrophic factors providing developmental support of dopamine neurons were unknown, but there is now growing evidence that glial-cell-line-derived neurotrophic factor (GDNF) serves as a physiologic limiting neurotrophic factor for these neurons during the first phase of natural cell death. During this phase, intrastriatal injection of GDNF diminishes the natural cell death event and neutralizing antibodies augment it. Sustained overexpression of GDNF in the striatum throughout development in a unique double transgenic mouse model results in an increased number of dopamine neurons surviving the first phase of natural cell death. However, this increase does not persist into adulthood. Therefore, other factors or mechanisms must play important roles in the determination of the mature number of nigral dopamine neurons. Further elucidation of these mechanisms will be important in the development of neuroprotective and cell replacement therapies for Parkinson's disease. PMID- 15349768 TI - Telomerase RNA deficiency in peripheral blood mononuclear cells in X-linked dyskeratosis congenita. AB - Compromised renewal and eventual failure of the hematopoietic system in dyskeratosis congenita (DC) have been proposed to arise from a deficiency in telomerase function. Previously, cultured cell lines from patients with X-linked DC were shown to accumulate less telomerase RNA than cell lines from unaffected family members. Here, we report that telomerase RNA deficiency is also present in the circulating lymphocytes of DC patients. We have compared the accumulation levels of telomerase RNA and a panel of other small RNAs in peripheral blood mononuclear cells from an X-linked DC patient and an unaffected maternal carrier and similarly analyzed cultured lymphoblasts from an X-linked DC patient and maternal carrier in a second family. The DC-patient lymphoid cells show a specific defect in telomerase RNA accumulation with or without cell culture. Our findings support the clinical significance of telomerase deficiency and encourage the use of telomerase activation as a disease therapy. PMID- 15349769 TI - Positive selection in MAOA gene is human exclusive: determination of the putative amino acid change selected in the human lineage. AB - Monoamine oxidase A (MAOA) is the X-linked gene responsible for deamination and subsequent degradation of several neurotransmitters and other amines. Among other activities, the gene has been shown to play a role in locomotion, circadian rhythm, and pain sensitivity and to have a critical influence on behavior and cognition. Previous studies have reported a non-neutral evolution of the gene attributable to positive selection in the human lineage. To determine whether this selection was human-exclusive or shared with other species, we performed a population genetic analysis of the pattern of nucleotide variation in non-human species, including bonobo, chimpanzee, gorilla, and orangutan. Footprints of positive selection were absent in all analyzed species, suggesting that positive selection has been recent and unique to humans. To determine which human-unique genetic changes could have been responsible for this differential evolution, the coding region of the gene was compared between human, chimpanzee, and gorilla. Only one human exclusive non-conservative change is present in the gene: Glu151Lys. This human substitution affects protein dimerization according to a three-dimensional structural model that predicts a non-negligible functional shift. This is the only candidate position at present to have been selected to fixation in humans during an episode of positive selection. Divergence analysis among species has shown that, even under positive selection in the human lineage, the MAOA gene did not experience accelerated evolution in any of the analyzed lineages, and that tools such as K(a)/ K(s) would not have detected the selective history of the gene. PMID- 15349771 TI - Nonsense mutations in the essential gene SUP35 of Saccharomyces cerevisiae are non-lethal. AB - In the present work we have characterized for the first time non-lethal nonsense mutations in the essential gene SUP35, which codes for the translation termination factor eRF3 in Saccharomyces cerevisiae. The screen used was based on selection for simultaneous suppression of two auxotrophic nonsense mutations. Among 48 mutants obtained, sixteen were distinguished by the production of a reduced amount of eRF3, suggesting the appearance of nonsense mutations. Fifteen of the total mutants were sequenced, and the presence of nonsense mutations was confirmed for nine of them. Thus a substantial fraction of the sup35 mutations recovered are nonsense mutations located in different regions of SUP35, and such mutants are easily identified by the fact that they express reduced amounts of eRF3. Nonsense mutations in the SUP35 gene do not lead to a decrease in levels of SUP35 mRNA and do not influence the steady-state level of eRF1. The ability of these mutations to complement SUP35 gene disruption mutations in different genetic backgrounds and in the absence of any tRNA suppressor mutation was demonstrated. The missense mutations studied, unlike nonsense mutations, do not decrease steady-state amounts of eRF3. PMID- 15349770 TI - SCH9, a putative protein kinase from Saccharomyces cerevisiae, affects HOT1 stimulated recombination. AB - HOT1 is a mitotic recombination hotspot derived from yeast rDNA. To further study HOT1 function, trans-acting H OT1 recombination mutants (hrm) that alter hotspot activity were isolated. hrm2-1 mutants have decreased HOT1 activity and grow slowly. The HRM2 gene was cloned and found to be identical to SCH9, a gene that affects a growth-control mechanism that is partially redundant with the cAMP dependent protein kinase A (PKA) pathway. Deletion of SCH9 decreases HOT1 and rDNA recombination but not other mitotic exchange. Although high levels of RNA polymerase I transcription initiated at HOT1 are required for its recombination stimulating activity, sch9 mutations do not affect transcription initiated within HOT1. Thus, transcription is necessary but not sufficient for HOT1 activity. TPK1, which encodes a catalytic subunit of PKA, is a multicopy suppressor of the recombination and growth defects of sch9 mutants, suggesting that increased PKA activity compensates for SCH9 loss. RAS2( val19), which codes for a hyperactive RAS protein and increases PKA activity, suppresses both phenotypic defects of sch9 mutants. In contrast to TPK1 and RAS2(val19), the gene for split zinc finger protein 1 (SFP1) on a multicopy vector suppresses only the growth defects of sch9 mutants, indicating that growth and HOT1 functions of Sch9p are separable. Sch9p may affect signal transduction pathways which regulate proteins that are specifically required for HOT1-stimulated exchange. PMID- 15349772 TI - Transferrin receptor induction in Toxoplasma gondii-infected HFF is associated with increased iron-responsive protein 1 activity and is mediated by secreted factors. AB - Infection with the apicomplexan parasite Toxoplasma gondii results in a significant alteration of the host-cell transcriptional profile. We have previously shown that the transferrin receptor (TfR) is specifically up-regulated in T. gondii-infected human fibroblasts but not in host cells infected with the bacterial pathogens Salmonella Typhimurium and Chlamydia trachomatis. In this report, we describe the prerequisites and physiological conditions that are associated with this pathogen-specific gene induction. Band-shift assays revealed that T. gondii infection resulted in increased activity in the iron response protein IRP1, which, in this state, stabilizes TfR mRNA from degradation. Although T. gondii depends on host-cell iron as demonstrated by sensitivity to deferoxamine, a parasite-induced iron starvation is not responsible for TfR up regulation. The increased iron availability due to treatment with holotransferrin and FeNTA did not prevent TfR induction nor was the transferrin-independent iron transporter NRAMP2 up-regulated in infected host cells. In addition, inhibition of parasite replication by drug treatment did not prevent TfR up-regulation. Instead, TfR induction was sensitive to cycloheximide and could be induced by treatment with conditioned media from infected human fibroblasts. Together our findings suggest that the T. gondii-specific TfR up-regulation is not due to a direct interaction of parasitic factors with the iron-uptake machinery of the host cell but is instead mediated indirectly as a result of secreted host cell- or parasite-derived factors. PMID- 15349773 TI - Monoclonal antibodies identify two neutralization-sensitive epitopes in Besnoitia besnoiti endocytes. AB - Four monoclonal antibodies were produced against endozoite membrane and cytoplasmic antigens of B. besnoiti. In immunofluorescence antibody tests, three of the clones, designated 2M3C5, 2M1G8 and 2M9G3 recognized antigens restricted to the anterior pole of the endozoites. The fourth clone, 2M9C4, recognized a membrane-associated component in a "beaded" pattern, cytoplasmic granules and extracellular background. The staining characteristics differed from the solid diffuse staining of polyclonal serum. On Western blots of detergent-soluble extracts fractionated under non-reducing conditions in 10% SDS-PAGE gels, mAbs 2M3C5, 2M1G8 and 2M9G3 recognized a common antigen at >200 kDa. Recognition with mAb 2M3C5 was consistently different in intensity and extent. Monoclonal antibody 2M9C4 recognized a single antigen at 75 kDa. The antibodies significantly reduced infectivity of Besnoitia endozoites into cultured cells, demonstrating the potential role of the antigens in the invasion process and raising the possibility of development of a vaccine and diagnostic tests for the disease. PMID- 15349774 TI - Giardia lamblia: behavior of the nuclear envelope. AB - Giardia Lamblia is a flagellar parasite possessing the unusual morphology of bearing two nuclei. New morphological observations on trophozoites and encysting Giardia nuclei using routine transmission electron microscopy, freeze fracture and cytochemistry are presented. Nuclear pores of both nuclei in the same cells were assessed on freeze-fracture replicas from different cell cycle phases, and compared. These techniques showed that (1) both nuclei in the same cell are distinct in nuclear pore number and distribution; (2) nuclear pore complexes are frequently clustered in nuclear envelope domains; (3) dividing nuclei display very few nuclear pores; (4) few ribosomes are found on the outer nuclear envelope of the trophozoite form; (5) nuclear membranes present spots of closely apposed membranes, which are different from the typical diaphragm nuclear pore complexes; (6) in addition to the nuclear pores, membrane blebs are also present in the nuclear envelope; (7) encysting cells show intranuclear inclusions, morphologically similar to the ESV (encystation-specific vesicles) and to the ER membranes, which may be the result of nuclear envelope folding. It is proposed that the two nuclei in Giardia are dissimilar in morphology and activity. PMID- 15349775 TI - Bioterrorism: is it a real threat? AB - The Geneva Protocol of 1925 commits the signatory nations to refraining from the use of biological weapons. However, the terrorist assaults of September 2001 and, subsequently, the anthrax-containing letters are cause for great concerns: new threats to the security of nations are expected, as terrorist organizations seem to increasingly explore novel ways of spreading terror. In this context, naturally emerging diseases such as SARS, monkeypox or West Nile fever assume new importance because it is difficult to distinguish between natural epidemics and possible bioweapon assaults. Great efforts on the part of governments and public health authorities are necessary to counteract these threats. PMID- 15349776 TI - Serial position effects in implicit memory for multiple-digit numbers. AB - Serial position effects in implicit and explicit memory were investigated in a short-term memory task. A study list composed of four, spatially distributed, two digit numbers was presented, followed by an item recognition task (explicit test) and an implicit memory task in which participants were asked to verify a simple addition equation where the presented answer was either primed or not primed by one of the number pairs in the study list. Similar serial position effects were observed in explicit and implicit memory, with faster response times for correct decisions on the first than on the later list positions. The presence of a primacy effect but no recency effect is consistent with previous studies of explicit memory with visual presentation. The results suggest that similar principles of temporal information processing govern priming and episodic short term memory. PMID- 15349777 TI - On the necessity of new characters for classification and systematics of biotrophic Peronosporomycetes. PMID- 15349778 TI - Functional analysis of a Douglas-fir metallothionein-like gene promoter: transient assays in zygotic and somatic embryos and stable transformation in transgenic tobacco. AB - Douglas-fir (Pseudotsuga menziesii [Mirb] Franco) metallothionein (PmMT) cDNA encodes a novel cysteine- and serine-rich MT, indicating a new subtype or prototype MT from which other plant MTs may have evolved. A genomic library of Douglas-fir was screened using MT cDNA probes, and genomic sequences that mediate tissue-specific, temporal as well as inducible expression of the embryo-specific MT-gene were analyzed. The promoter region of the PmMT genomic clone (gPmMT) contained a hexameric G-box, two putative ethylene-responsive elements and an inverted repeat of a motif similar to the core metal regulatory element. Interestingly, comparison of the upstream region of Douglas-fir gPm2S1 and gPmMTa genes revealed a conserved motif, CATTATTGA, not found in any known angiosperm gene promoter. Chimeric gene constructs containing a series of deletions in the gPmMTa promoter fused to the uidA reporter gene were assayed in Douglas-fir and transgenic tobacco (Nicotiana tabacum L.). Transient-expression assays in Douglas fir megagametophyte and zygotic embryos indicated that the sequence -190 to +88 of gPmMTa was sufficient to drive the expression of the reporter gene and that the 225-bp fragment (-677 to -453) contained sequences necessary for high-level expression. In transgenic tobacco seedlings the beta-glucuronidase activity was localized in the vacuolar tissue and proliferating tissue of the auxiliary buds and stem elongation zone. The gPmMTa promoter was not active in the seeds of transgenic tobacco or in the roots of seedlings up to 3 weeks old. Detailed studies of transient expression and stable transformation provided important information on evolutionary conservation as well as novel features found in the conifer promoter. This is the first report of an MT-like gene promoter from conifers. PMID- 15349780 TI - An auxin-responsive 1-aminocyclopropane-1-carboxylate synthase is responsible for differential ethylene production in gravistimulated Antirrhinum majus L. flower stems. AB - The regulation of gravistimulation-induced ethylene production and its role in gravitropic bending was studied in Antirrhinum majus L. cut flower stems. Gravistimulation increased ethylene production in both lower and upper halves of the stems with much higher levels observed in the lower half. Expression patterns of three different 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) genes, an ACC oxidase (ACO) and an ethylene receptor (ETR/ERS homolog) gene were studied in the bending zone of gravistimulated stems and in excised stem sections following treatment with different chemicals. One of the ACS genes (Am-ACS3) was abundantly expressed in the bending zone cortex at the lower side of the stems within 2 h of gravistimulation. Am-ACS3 was not expressed in vertical stems or in other parts of (gravistimulated) stems, leaves or flowers. Am-ACS3 was strongly induced by indole-3-acetic acid (IAA) but not responsive to ethylene. The Am-ACS3 expression pattern strongly suggests that Am-ACS3 is responsible for the observed differential ethylene production in gravistimulated stems; its responsiveness to IAA suggests that Am-ACS3 expression reflects changes in auxin signalling. Am ACS1 also showed increased expression in gravistimulated and IAA-treated stems although to a much lesser extent than Am-ACS3. In contrast to Am-ACS3, Am-ACS1 was also expressed in non-bending regions of vertical and gravistimulated stems and in leaves, and Am-ACS1 expression was not confined to the lower side cortex but evenly distributed over the diameter of the stem. Am-ACO and Am-ETR/ERS expression was increased in both the lower and upper halves of gravistimulated stems. Expression of both Am-ACO and Am-ETR/ERS was responsive to ethylene, suggesting regulation by IAA-dependent differential ethylene production. Am-ACO expression and in vivo ACO activity, in addition, were induced by IAA, independent of the IAA-induced ethylene. IAA-induced growth of vertical stem sections and bending of gravistimulated flowering stems were little affected by ethylene or 1-methylcyclopropene treatments, indicating that the differential ethylene production plays no pivotal role in the kinetics of gravitropic bending. PMID- 15349781 TI - Cold acclimation in bryophytes: low-temperature-induced freezing tolerance in Physcomitrella patens is associated with increases in expression levels of stress related genes but not with increase in level of endogenous abscisic acid. AB - Bryophyte species growing in areas in which temperatures fall below zero in winter are likely to have tolerance to freezing stress. It is well established in higher plants that freezing tolerance is acquired by exposure to non-freezing low temperatures, accompanied by expression of various genes and increases in levels of the stress hormone abscisic acid (ABA). However, little is known about the physiological changes induced by cold acclimation in non-vascular plants such as bryophytes. We examined the effects of low temperatures on protonema cells of the moss Physcomitrella patens (Hedw.) Bruch and Schimp. The freezing tolerance of protonema cells was clearly increased by incubation at low temperatures ranging from 10 degrees C to 0 degrees C, with maximum tolerance achieved by incubation at 0 degrees C for several days. The enhancement of freezing tolerance by low temperatures occurred in both light and dark conditions and was accompanied by accumulation of several transcripts for late-embryogenesis-abundant (LEA) proteins and boiling-soluble proteins. By de-acclimation, low-temperature-induced expression of these transcripts and proteins, as well as the freezing tolerance, was reduced. Interestingly, endogenous levels of ABA in tissues or that secreted into the culture medium were not specifically increased by low-temperature treatment. Furthermore, removal of ABA from the medium by addition of activated charcoal did not affect low-temperature-induced freezing tolerance of the protonema cells. Our results provide evidence that bryophytes have an ABA independent cold-signaling pathway leading to expression of stress-related genes and resultant acquisition of freezing tolerance. PMID- 15349782 TI - Comparison of computerized methods for detecting the ventilatory thresholds. AB - The aim of this study was to compare computerized automatic methods to detect the ventilatory threshold (VT). Thirty apparently healthy and physically active volunteers [22.5 (6.5) years; 1.72 (0.08) m; 71.9 (8.5) kg] were submitted to a progressive and maximal cycle exercise. The gas exchange was monitored breath-by breath with a fast gas analyser. The VT and respiratory compensation (RC) were automatically detected based on the respiratory exchange ratio, the ventilatory equivalent for O2 and the ventilatory equivalent for CO2, pulmonary ventilation, end-tidal PO2 and PCO2, and v-slope. In addition, VT and RC were also determined independently by visual inspection by two experienced investigators, and the results were compared with those of the automatic procedures. The automatic VT averaged 77% of the maximal VO2 and the RC 88%. The agreement between the experienced observers was very close [mean difference: 44.4 (16.1) ml, r = 0.94, not significant]. Data were expressed as the mean value together with the standard deviation in each case. The automatic and visual inspection procedures did not present significant differences, resulting in 29.6 (29.6) ml with a reliability of r = 0.86. All methods were significantly correlated for VT and RC (r = 0.93 on average, P < 0.01). ANOVA did not show differences between either the VT methods (P = 0.131) or the RC methods (P = 0.41). In conclusion, the present study has compared several simultaneous breath-by-breath ergospirometric methods that are used to describe the anaerobic threshold, showing high confidence when compared to visual inspection. No statistical differences were found between the VT and RC techniques for physically active subjects indicating that these methods may be equally effectively employed. PMID- 15349783 TI - Excess post-exercise oxygen consumption in spinal cord-injured men. AB - This study examined excess post-exercise oxygen consumption (EPOC) following arm cranking in men who had a traumatic spinal cord injury (SCI). Six physically active SCI men with a lesion level between T10 and T12 and six able-bodied (AB) men who were matched according to upper body peak VO(2) performed 30 min of arm cranking at 65-70% peak VO(2). Baseline measurements were recorded during the last 10 min of a 40-min seated rest. Subjects remained seated during recovery for 40 min or until VO(2) returned to baseline, whichever was longer. Plasma lactate concentration was measured at rest, at the end of exercise, and at 10, 20 and 40 min of recovery. EPOC duration was not significantly different ( P>0.05) between SCI [23.2 (7.9) min; mean (SE)] and AB [35.0 (15.4) min] men, nor was there a significant group difference in EPOC magnitude [36.8 (7.8) kJ for SCI and 53.0 (22.8) kJ for AB]. There was no significant difference in recovery heart rate (HR) or respiratory exchange ratio (RER) between SCI and AB. However, HR measured at the end of the EPOC period was significantly elevated ( P<0.001) and RER significantly lower ( P<0.03) for both groups when compared to baseline. Lactate concentration was not significantly different between the groups at any sampling period. The findings suggest that physically active SCI men have a similar energy expenditure and time frame for recovery from arm crank exercise as their AB counterparts. Similar to what has been reported following lower body exercise, arm crank exercise elicits a higher HR and lower RER at end-EPOC when compared to pre-exercise values. PMID- 15349784 TI - The effect of acute branched-chain amino acid supplementation on prolonged exercise capacity in a warm environment. AB - Eight males were recruited to examine the effect of branched-chain amino acid (BCAA) supplementation on exercise capacity in a glycogen-depleted state in a warm environment. Following a exercise and dietary regimen designed to reduce glycogen availability, subjects returned to the laboratory the following morning and remained seated for 2 h, before cycling to volitional exhaustion at 50% VO2 peak in a warm environment [30.0 (0.2) degrees C; mean (SD)]. Four 250 ml aliquots of a 12 g l(-1) BCAA solution or placebo were ingested at 30 min intervals prior to exercise, with an additional 150 ml consumed every 15 min throughout exercise. BCAA ingestion had no effect on exercise capacity [placebo 103.9 (26.9) min; BCAA 111.0 (29.2) min; P = 0.129). No difference in heart rate (P = 0.345), core temperature (P = 0.628), or weighted mean skin temperature (P = 0.114) was apparent between trials. Ingestion of the BCAA solution produced a marked increase in plasma BCAA immediately prior to exercise [+ 1126 (158) micromol l(-1); P < 0.001) with this difference maintained throughout. Consequently, a significant reduction in the plasma concentration ratio of free tryptophan to BCAA was observed during the BCAA trial when compared to the placebo (P < 0.001). Plasma ammonia concentration was significantly elevated during exercise throughout the BCAA trial (P < 0.001), with no change from rest apparent during the placebo trial (P = 0.608). Blood glucose (P = 0.114) and lactate (P = 0.836) concentrations were not different between trials. Ingestion of a BCAA solution prior to, and during, prolonged exercise in glycogen-depleted subjects did not influence exercise capacity in a warm environment. PMID- 15349786 TI - Treatment of primary intraocular lymphoma with oral trofosfamide: report of two cases and review of the literature. AB - BACKGROUND: The prognosis of primary intraocular lymphoma (PIOL) is poor. Due to frequent ocular and/or CNS relapses, the optimal treatment for PIOL has not yet been defined. This is the first report of PIOL treatment with trofosfamide. METHODS: The first patient with a newly diagnosed PIOL was not considered eligible for high-dose methotrexate-based systemic chemotherapy due to comorbidities. The second patient had a primary central nervous system lymphoma, which was resistant to three chemotherapy regimens and responded to whole-brain irradiation. A tumor relapse in the eye was treated with local radiotherapy with initial success; however, the tumor subsequently recurred. The first patient received oral trofosfamide 150 mg daily, the second 400 mg daily (reduced after 5 months of therapy to 300 mg daily) over 5 consecutive days, followed by a 5-day drug-free interval. RESULTS: In both patients, complete remission was achieved, which lasted for 8+ and 18 months, respectively. Except for a temporary asymptomatic decrease in hemoglobin in the first patient, no toxicity was observed. CONCLUSIONS: Trofosfamide may offer an alternative treatment option for PIOL with a very favorable side effect profile. The exact role of trofosfamide in the treatment of PIOL remains to be defined. PMID- 15349787 TI - The labeling efficiency of human telomeres is increased by double-strand PRINS. AB - Telomeres are composed of tandem repeated sequences, TTAGGG, that can be detected either by fluorescence in situ hybridization (FISH), more efficiently by using a peptide nucleic acid (PNA) probe, or by the primed in situ (PRINS) technique. However, the efficiency of human telomere labeling using PRINS is somewhat lower than the efficiency using PNA-FISH. To solve this problem, we developed a double strand PRINS technique, which uses two primers, (TTAGGG)(7) and (CCCTAA)(7), to label both forward and reverse telomeric DNA strands. A total of 120 lymphocyte metaphases obtained from three normal adults were scored to evaluate the labeling efficiency based upon the telomere signal frequency present in chromatid ends and chromosome arms. As a comparison, 30 metaphases from the same three individuals were evaluated using PNA-FISH. The average labeling efficiency of PRINS was increased to a level very close to that obtained with PNA-FISH. Therefore, we demonstrated that the low labeling efficiency of human telomeres with regular PRINS was likely caused by uneven annealing of primers at the relatively short human telomere sequences, resulting in some telomere sites with very weak or absent labeling. We suggest that the present double-strand labeling protocol is critical to maximize the labeling efficiency of the human telomere sequence when using the PRINS technique. PMID- 15349788 TI - SUMO modified proteins localize to the XY body of pachytene spermatocytes. AB - The XY body is a specialized chromatin territory that forms during meiotic prophase of spermatogenesis and comprises the transcriptionally repressed sex chromosomes. Remodeling of the XY chromatin is brought about by recruitment of specific proteins to the X and Y chromosomes during meiosis, and also by post translational modifications of histones and other chromatin-associated proteins. Here, we demonstrate that SUMO, a small ubiquitin-related modifier protein that regulates a wide variety of nuclear functions in somatic cells, dramatically localizes to the XY body. SUMO was first detected in the XY body of early pachytene spermatocytes and gradually accumulated, reaching maximal levels there during the mid to late pachytene stages. Several known SUMO substrates, including PML and DAXX, were also found to accumulate in the XY body of mid to late stage pachytene spermatocytes. These same proteins localize to PML nuclear bodies of somatic interphase nuclei. Together, these findings indicate a role for SUMO modification in regulating the structure and function of the XY body and reveal molecular similarities between the XY body and PML nuclear bodies. PMID- 15349789 TI - A simple reconstructed human epidermis: preparation of the culture model and utilization in in vitro studies. AB - The preparation of a reconstructed human epidermis is described with examples of its utilization in in vitro studies. The model was obtained by culturing normal human keratinocytes at high cell density for 14 days in serum-free and high calcium (1.5 m M) medium on an inert polycarbonate filter at the air-liquid interface. These stratified cultures showed histological features similar to those observed in vivo in the epidermis: a proliferating basal layer and differentiating spinous, granular, and cornified layers. Electron microscopy illustrated lamellar bodies, junctions and keratohyalin granules. Immunofluorescent localization of epidermal markers (keratins 14 and 10, involucrin and filaggrin) revealed typical differentiation. This in vitro reconstructed tissue was used in studies of toxic effects of chemicals. The modelled tissue showed progressive cytotoxicity of a skin irritant (benzalkonium chloride) and a sensitizer (dinitrochlorobenzene) as assessed by MTT assay. Moreover, differential release of interleukin-1alpha and interleukin-8 were measured after 20 h of incubation allowing the irritant to be distinguished from the sensitizer. Permeation studies indicated efficient barrier function of the reconstructed epidermis, as well as metabolizing properties towards hormones. This model can be custom-made and is potentially useful for studies involving keratinocytes in the epidermis, in basic science, dermatology or toxicology. PMID- 15349790 TI - Why is polymorphous light eruption so common in young women? AB - Despite the fact that polymorphous light eruption (PLE) is the most common photodermatosis, affecting 15% of healthy people in the UK, its pathogeny remains unclear. The condition is more frequent in females and begins often in young adults and in mid-adult life. The mechanism of PLE is under active research as shown by recent results, and it is hypothesized that in PLE patients, there is a partial failure of ultraviolet radiation-induced immunosuppression, causing an abnormal response to autologous antigens generated by ultraviolet radiation (UVR). The recent demonstration that the female hormone, 17beta-estradiol prevents UVR-induced suppression of the contact hypersensitivity response caused by the release of immunosuppressive cytokines (IL-10) from keratinocytes might thus explain why the risk of PLE is higher in females than in males and why the risk decreases in women after the menopause. PMID- 15349791 TI - Abstracts of ESMRMB 2004, 21st Annual Scientific Meeting of the European Society for Magnetic Resonance in Medicine and Biology. Copenhagen, Denmark. September. 9 12, 2004. PMID- 15349792 TI - Influence of food dispersion on feeding activity and social interactions in captive Lophocebus albigena and Cercocebus torquatus torquatus. AB - We analysed the impact of the distribution of food items on feeding activity and social interactions in captive red-capped and grey-cheeked mangabeys. Three different feeding situations were presented: food items were either placed in a single heap or dispersed in several smaller heaps on the ground or in space. Social interactions were estimated by the frequency of positive, as well as of negative social interactions and by the structure of visual social attention. Feeding activity was estimated by proximity to food and feeding frequency. When food items were presented in single heaps, the adult males monopolized the food and monitoring of conspecifics increased. Social interactions and social gazes decreased in numbers when food items were dispersed in 3D. Gazes were directed more frequently towards the adult males when food items were presented in a single heap. Juveniles and some nonreproductive adult monkeys were the most affected by food competition situations. Inter-group variations, within a given species, of the behavioural responses observed in relation to the distribution of food items were evidenced. This suggests that social context, i.e., individual histories and relationship between group members, plays an important part in the expression of the activities of each member in a group. PMID- 15349793 TI - Grooming relationships of adolescent orphans in a free-ranging group of Japanese macaques (Macaca fuscata) at Katsuyama: a comparison among orphans with sisters, orphans without sisters, and females with a surviving mother. AB - The present study examined grooming relationships of adolescent females in a free ranging group of Japanese macaques (Macaca fuscata) at Katsuyama. To assess whether the loss of the mother influenced the grooming relationships of adolescent females (5-7 years old), we compared the time spent in grooming interactions and the number of grooming partners among the following three groups: 6 adolescent orphans with sisters, 9 adolescent orphans without sisters, and 11 adolescent non-orphans with surviving mothers. In Japanese macaques, grooming most frequently occurs between mothers and their daughters. Therefore, it is expected that if the mother is lost, orphans will devote less time to grooming interactions than non-orphans. However, the time spent in overall grooming interactions did not differ among the three groups. While non-orphans maintained grooming relationships with their mothers, orphans acquired alternative grooming relationships with other group members. Orphans adopted two kinds of tactics to compensate for the loss of the mother. First, adolescent orphans with sisters developed more affiliative grooming relationships with their sisters than non-orphans with sisters. Secondly, adolescent orphans without sisters spent more time in grooming interactions with same-aged females and non related adult females. Moreover, regarding grooming interactions with same-aged females and non-related adult females, orphans without sisters had a larger number of grooming partners than non-orphans. These results indicate that adolescent females have enough flexibility to develop their grooming network after the loss of their mothers, and that the lack of mother and sisters might accelerate socialization of adolescent females and enable them to be integrated in reciprocal adult grooming relationships. PMID- 15349794 TI - A comparative genomic analysis of ESTs from Ustilago maydis. AB - A large-scale comparative genomic analysis of unisequence sets obtained from an Ustilago maydis EST collection was performed against publicly available EST and genomic sequence datasets from 21 species. We annotated 70% of the collection based on similarity to known sequences and recognized protein signatures. Distinct grouping of the ESTs, defined by the presence or absence of similar sequences in the species examined, allowed the identification of U. maydis sequences present only (1) in fungal species, (2) in plants but not animals, (3) in animals but not plants, or (4) in all three eukaryotic lineages assessed. We also identified 215 U. maydis genes that are found in the ascomycete but not in the basidiomycete genome sequences searched. Candidate genes were identified for further functional characterization. These include 167 basidiomycete-specific sequences, 58 fungal pathogen-specific sequences (including 37 basidiomycete pathogen-specific sequences), and 18 plant pathogen-specific sequences, as well as two sequences present only in other plant pathogen and plant species. PMID- 15349795 TI - Laser acupuncture: past, present, and future. AB - Laser acupuncture is defined as the stimulation of traditional acupuncture points with low-intensity, nonthermal laser irradiation. Although the therapeutic use of laser acupuncture is rapidly gaining in popularity, objective evaluation of its efficacy in published studies is difficult because treatment parameters such as wavelength, irradiance, and beam profile are seldom fully described. The depth of laser energy transmission, likely an important determinant of efficacy, is governed not only by these parameters, but also by skin properties such as thickness, age, and pigmentation-factors which have also received little consideration in laser acupuncture. Despite the frequently equivocal nature of the published laser studies, recent evidence of visual cortex activation by laser acupuncture of foot points, together with the known ability of laser irradiation to induce cellular effects at subthermal thresholds, provides impetus for further research. PMID- 15349796 TI - Superior vena cava syndrome: a rare clinical manifestation of Behcet's disease. PMID- 15349797 TI - Proximal myopathy as an unusual presenting feature of celiac disease. AB - A 37-year-old woman presented with back pain, diffuse musculoskeletal pain, and muscle weakness without marked gastrointestinal symptoms. She complained of difficulty in walking and bilateral hip pain for the preceding year. Clinical examination revealed proximal muscle weakness especially in the lower extremities and a waddling gait pattern. Laboratory parameters and radiographic findings revealed the diagnosis of osteomalacia. The etiology of osteomalacia was investigated and a diagnosis of celiac disease was established. As osteomalacia symptoms may be the only presenting feature of celiac disease, it should be considered in the differential diagnosis of patients presenting with proximal muscle weakness and diffuse musculoskeletal pain. PMID- 15349798 TI - Soluble adhesion molecules (sVCAM-1, sE-selectin), vascular endothelial growth factor (VEGF) and endothelin-1 in patients with systemic sclerosis: relationship to organ systemic involvement. AB - Systemic sclerosis (SSc) is a chronic, multisystemic, autoimmune disease characterised by vascular changes and varying degrees of fibrosis of the skin and visceral organs. Organ systemic involvement in SSc is associated with an altered function of endothelial cells, perivascular infiltrating mononuclear cells and interstitial fibrosis. To evaluate the relationship between systemic manifestations and immunological markers of endothelial cell activation, serum levels of soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble E-selectin (sE-selectin), vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) were determined by an enzyme-linked immunosorbent assay in 31 SSc patients and in 30 healthy controls. In comparison with the control group, higher serum concentrations of sVCAM-1, sE-selectin, VEGF and ET-1 were detected in SSc patients (in all cases p<0.001). Elevated concentrations of sVCAM-1 (p<0.05), sE selectin (p<0.05), VEGF (p<0.05) and ET-1 (p<0.01) dominated in the serum of SSc patients with organ systemic involvement compared to those without systemic manifestation of the disease. These results suggest that the serum levels of sVCAM-1, sE-selectin, VEGF and ET-1 may reflect the extent of internal organ involvement in SSc patients and point to a pathogenic role of these molecules in systemic manifestation of the disease. PMID- 15349799 TI - Glucose-6-phosphate dehydrogenase (G6PD) mutations in Myanmar: G6PD Mahidol (487G>A) is the most common variant in the Myanmar population. AB - We conducted a survey of malaria diagnoses and treatments in remote areas of Myanmar. Blood specimens from more than 1,000 people were collected by the finger prick method, and 121 (11%) of these people were found to be glucose-6-phosphate dehydrogenase (G6PD) deficient. Of these 121, 50 consented to analysis of the G6PD genome. We read the G6PD sequences of these subjects and found 45 cases of G6PD Mahidol (487G>A), two of G6PD Coimbra (592C>T), two of G6PD Union (1360C>T), and one of G6PD Canton (1376G>T). Taken together with data from our previous report, 91.3% (73/80) of G6PD variants were G6PD Mahidol. This finding suggests that the Myanmar population is derived from homogeneous ancestries and are different from Thai, Malaysian, and Indonesian populations. PMID- 15349801 TI - Epidemiology of alcoholic liver disease. AB - Alcohol is one of the main causes of end-stage liver disease worldwide, and alcoholic liver disease is the second most common reason for liver transplantation in the United States. Beginning in the 1970s, there was a gradual decline in alcoholic cirrhosis-related mortality in many countries. However, in the past few years, alcoholic liver disease mortality rates in several countries have stabilized or started to increase. There are significant ethnic and gender differences in alcoholic cirrhosis-related mortality rates. Furthermore, alcohol use increases the risk for liver disease in those infected with hepatitis C. A better understanding of the epidemiology of alcoholic liver disease will allow for improved diagnosis and management of this common problem. PMID- 15349802 TI - Diagnosis and therapy of alcoholic liver disease. AB - Alcoholic liver disease (ALD) presents considerable challenges to clinicians. Screening for alcohol abuse and alcoholism should be routine and repeated annually with close attention to signs and symptoms of liver disease. In patients with evidence of liver dysfunction or injury, consideration should be given to performance of liver biopsy for diagnosis and prognosis and prior to initiation of medication with the potential for significant side effects. Therapy depends on the spectrum of pathological liver injury: alcoholic fatty liver, alcoholic hepatitis, and cirrhosis. Abstention is the foundation of therapy for an alcohol problem. Alcoholic fatty liver should improve with abstention, but the similarity to the pathogenesis of nonalcoholic fatty liver and potential for progressive injury merits consideration of lipotropic agents. The continuing mortality, poor acceptance of corticosteroids, and identification of tumor necrosis factor-alpha (TNF-alpha) as an integral component has led to studies of pentoxifylline and, recently, anti-TNF antibody to neutralize cytokines in the therapy of severe alcoholic hepatitis. Antioxidant therapy of alcoholic cirrhosis has significant promise but will require large clinical trials. PMID- 15349803 TI - Transplantation in the alcoholic patient. AB - Alcoholic liver disease (ALD) is the second leading indication for transplantation in the United States. Most transplant programs in the United States require a minimum of 6 month's abstinence before transplantation is performed. Most studies have shown a recidivism rate of between 20 and 30% by 2 years after orthotopic liver transplantation (OLT). Higher rates of recidivism are reported if pre-OLT abstinence was < 6 months. The impact of recidivism on patient and graft survival is not clear because most reports include patients who consume alcohol in small amounts or infrequently. Equal post-OLT survival for ALD patients and non-ALD patients has been demonstrated, and ALD patients are not thought to suffer greater morbidity post transplant than non-ALD patients. Careful pretransplant assessment for concomitant medical and psychosocial ailments associated with alcoholism is important. Posttransplant monitoring for cardiovascular disease and withdrawal syndromes is required in the early postoperative setting, whereas monitoring for recidivism and malignancy are late postoperative issues. PMID- 15349804 TI - Cellular signaling mechanisms in alcohol-induced liver damage. AB - Chronic excessive alcohol intake is associated with multiple liver defects ranging from mild steatosis to advanced cirrhosis. However, the mechanisms by which chronic ethanol intake affects liver function remain a matter of intense debate and investigation. The liver is the major site of ethanol metabolism in the body, and a wide range of metabolic alterations is associated with ethanol intake. As a result, the liver is exposed to dramatic changes in redox state, transient hypoxia, episodes of oxidative stress, and the products of ethanol metabolism, such as acetaldehyde, acetate, and fatty acid ethyl esters. Chronic ethanol consumption is associated with increased levels of circulating endotoxins and proinflammatory cytokines that affect liver function. A major source of the increase in circulating proinflammatory cytokines is the Kupffer cells, which are sensitized to generate tumor necrosis factor alpha (TNF-alpha) through multiple mechanisms. In addition, the hepatocytes themselves are more susceptible to external stress. In isolated hepatocytes, this effect of chronic ethanol is evident in a greater sensitivity to proapoptotic challenges and, more specifically, to the cytotoxic actions of TNF-alpha. The mechanism by which hepatocytes are sensitized to external stress remains poorly characterized but may involve defects in mitochondrial function and oxidative defense mechanisms, the activation of death-promoting signaling pathways, and the inactivation of survival pathways. In this article, we emphasize the role of the stress-activated mitogen-activated protein kinase (MAPK) cascades in the onset of cell injury and their regulation by the phosphoinositide-3-kinase/Akt signaling cascade, which appears to function as the central integrating module of the stress-signaling machinery in the cell. We also discuss the complications and challenges of extrapolating these findings to the conditions in vivo and what we can learn from these studies regarding the nature of the liver defects associated with chronic alcohol consumption. PMID- 15349805 TI - Immunologic mechanisms of alcoholic liver injury. AB - Clinically, the association of alcoholic liver disease (ALD) with circulating autoantibodies, hypergammaglobulinemia, antibodies to unique hepatic proteins, and cytotoxic lymphocytes reacting against autologous hepatocytes strongly suggests altered immune regulation with an increased activity toward normal self proteins (loss of tolerance). Experimentally, there are several immune responses generated specifically recognizing self-proteins that are modified by metabolites of alcohol. These data strongly suggest that immune reactions may play a significant role in inducing and sustaining an inflammatory cascade of tissue damage to the liver. Additional support for this comes from the observation that the histological appearance of livers with ALD is that of a chronic active hepatitis-like inflammatory disease. Therefore, the hypothesis that immune mechanisms are involved in recurrent alcoholic hepatitis, although not summarily proven, is reasonable, supported by clinical and experimental evidence, and the subject of this article. PMID- 15349806 TI - Nutrition and alcoholic liver disease. AB - Malnutrition is a common finding in chronic alcoholics, and protein calorie malnutrition (PCM) is universal and predictive of survival in patients with established alcoholic liver disease (ALD). These patients also demonstrate frequent deficiencies of folate, thiamine, pyridoxine, and vitamin A, which enhance the likelihood of anemia, altered cognitive states, and night blindness. The etiologies of malnutrition in ALD patients are multiple and interactive and include anorexia with inadequate dietary intake, abnormal digestion of macronutrients and absorption of several micronutrients, increased skeletal and visceral protein catabolism, and abnormal interactions of ethanol and lipid metabolism. Numerous, and mostly inadequately controlled, studies have evaluated the potential efficacies of oral, enteral, and parenteral nutrition approaches to treatment of ALD, with mixed results on liver function, clinical improvements, and short- or long-term survival. Targeted metabolic treatments include supplementation with S-adenosylmethionine (SAM) or phosphatidylcholine derivatives, each with promising experimental bases but inconclusive clinical trials. PMID- 15349807 TI - Alcohol and hepatitis C. AB - Alcohol abuse and hepatitis C virus (HCV) infection coexist with chronic liver disease in many patients. The mechanism of injury in these patients is probably multifactorial and involves, but is not limited to, a combination of diminished immune clearance of HCV, oxidative stress, emergence of HCV quasi-species, hepatic steatosis, increased iron stores, and increased rate of hepatocyte apoptosis. In patients with HCV infection, alcohol consumption is known to cause accelerated progression of liver fibrosis, higher frequency of cirrhosis, and increased incidence of hepatocellular carcinoma (HCC). These patients also have decreased survival as compared with patients with either alcohol abuse or HCV liver injury alone. Alcohol abuse causes decreased response to interferon treatment in HCV patients. It is therefore necessary for patients with HCV infection to abstain from alcohol consumption. PMID- 15349808 TI - Alcohol and iron. AB - Iron in its free ferrous and ferric states may serve as a physiological regulator of normal intracellular functions but can be a double-edged sword when linked to several pathways of cellular toxicity. In particular, oxidative stress-induced cytotoxicity leading to both necrosis and apoptosis (so-called necrapoptosis) may be promoted by increased intracellular free iron. When hepatocyte iron accumulates to excess in clinical alcohol abuse or in an experimental, combined model of iron and alcohol hepatotoxicity, there is evidence for synergy among the putative pathways of oxidative stress. Just how excess hepatocyte iron accumulates in alcohol excess is unknown, but when the usual safe harbor for intracellular iron, namely the endosomal-lysosomal compartment, is compromised, it becomes a potent source of free, chelatable pro-oxidant iron. In this regard excess iron in alcohol-induced liver damage and alcohol excess in iron-overload disease are powerful cocktails promoting subcellular organelle damage leading to cell death and fibrogenesis. In some experimental circumstances, only the combined insults of both alcohol and iron overload are capable of inducing cell injury and fibrogenesis. There are clinical examples for such relative resistance to each hepatotoxin presented to the liver in isolation that lend support to this concept of synergy of pro-oxidant pathways of liver injury. PMID- 15349809 TI - A 48-year-old obese man with a 3 cm liver lesion and a history of hemochromatosis and lymphoma. PMID- 15349811 TI - Molecular cell biology and cancer metastasis. An interview with Garth Nicolson. PMID- 15349812 TI - Transcriptional regulation of cadherins during development and carcinogenesis. AB - The cadherin superfamily of Ca(2+)-dependent homophilic adhesion molecules plays a critical role in regulating cell-to-cell interactions. During development, the expression of different cadherins is highly dynamic, since they are associated with the morphogenesis, establishment and/or maintenance of different tissues. Alterations in cadherin expression or function occur frequently during carcinogenesis, such as the loss of the epithelial cadherin (E-cadherin) and/or the aberrant expression of other cadherins. Indeed, the aberrant expression of cadherins has been detected during carcinoma invasion, a process which is reminiscent of the epithelial-mesenchymal transition (EMT) so important in many critical developmental processes. The functional regulation of cadherins can occur at many different levels, from transcriptional regulation to the control of the strength of the cadherin-mediated cell-cell interaction. In this review, we will focus on the transcriptional control of cadherin expression, both in development and carcinogenesis, paying particular attention to the regulation of E-cadherin given its proposed role as a suppressor of invasion. We will discuss the main genetic and epigenetic mechanisms involved in down-regulating E-cadherin expression, and we will analyse the mechanisms involved in regulating EMT, in an attempt to elucidate which elements are common to this process in both physiological and pathological situations. PMID- 15349813 TI - APC dosage effects in tumorigenesis and stem cell differentiation. AB - It is well established that concentration gradients of signaling molecules (the so-called "morphogens") organize and pattern tissues in developing animals. In particular, studies in Drosophila and different vertebrates have shown that gradients of the Wnt, Hedgehog (Hh) and transforming growth factor-beta (TGF beta) families of morphogens play critical roles in limb patterning. Morphogens are often expressed in organizing centres and can act over a long range to coordinate the patterning of an entire field of cells. These observations imply that exposure to different concentrations of these diffusible factors may trigger differential cellular responses. In order to study these dosage-dependent Wnt/beta-catenin signaling effects, we have generated several hypomorphic mutant alleles at the mouse Apc locus and studied their cellular and phenotypic outcomes in stem cell renewal and differentiation, and in tumorigenesis. The results clearly show that Apc mutations differentially affect the capacity of stem cells to differentiate in a dosage-dependent fashion. Likewise, different Apc mutations (and the corresponding Wnt signaling dosages) confer different degrees of susceptibility to tumorigenesis in the corresponding mouse models. These results have implications for the understanding of the molecular and cellular basis of tumor initiation by defects in the Wnt pathway. We propose a model in which adult somatic stem cell compartments are characterized by tissue-specific beta-catenin threshold levels for cell proliferation, differentiation and apoptosis. Different APC mutations will result in different levels of beta-catenin signaling, thus conferring different degrees of tumor susceptibility in different tissues. Hence, beta-catenin dosage-dependent effects may not only explain how a single pathway is involved in the development and homeostasis of different tissues, but also its pleiotrophic role in tumorigenesis. PMID- 15349814 TI - Discovery and characterization of the cadherin family of cell adhesion molecules. An interview with Masatoshi Takeichi. PMID- 15349815 TI - Cadherin-mediated cell-cell adhesion and tissue segregation in relation to malignancy. AB - We review evidence concerning the basis for tissue segregation during embryonic development. This compartmentalization is shown to be an immiscibility phenomenon caused by changes in the strengths of adhesions between mobile cells which accompany their differentiation and generate interfacial tensions at cell population boundaries. The mobile cells exchange neighbors in response to these adhesion-generated forces which impel the system toward the configuration of maximal binding. Cadherins dominate these intercellular adhesions, but integrin fibronectin-based adhesions also contribute to them as well as to cell-matrix adhesions. At the interface between two segregating cell populations are three kinds of cell-cell interfaces: a-a, b-b and a-b. Tissue immiscibility (segregation) results when the cross-adhesion is weaker than the mean value of the two kinds of self-adhesions, does not require (although it permits) qualitative changes in cell adhesion molecules and is easily generated even by moderate changes in the quantities of adhesion molecules on the cell surfaces. All type I and II cadherins tested cross-adhere, in most cases with strengths close to those of their self-adhesions. Is malignant invasion a process of cell segregation in reverse, in which the cross-adhesion between cancer cells and host tissue components is strong relative to their self-adhesions? We review evidence for cadherin involvement in breast, prostate and brain cancers. Despite evidence that N-cadherin enhances the invasiveness of certain cancer cells, we have found that increasing the expression not only of functional E-cadherin but also of P- or N-cadherin restrains the spreading of other malignant cell lines over (and through) a reconstituted extracellular matrix. PMID- 15349816 TI - Matrix metalloproteinases in cancer: from new functions to improved inhibition strategies. AB - Over the last years, the relevance of the matrix metalloproteinase (MMP) family in cancer research has grown considerably. These enzymes were initially associated with the invasive properties of tumour cells, owing to their ability to degrade all major protein components of the extracellular matrix (ECM) and basement membranes. However, further studies have demonstrated the implication of MMPs in early steps of tumour evolution, including stimulation of cell proliferation and modulation of angiogenesis. The establishment of causal relationships between MMP overproduction in tumour or stromal cells and cancer progression has prompted the development of clinical trials with a series of inhibitors designed to block the proteolytic activity of these enzymes. Unfortunately, the results derived from using broad-spectrum MMP inhibitors (MMPIs) for treating patients with advanced cancer have been disappointing in most cases. There are several putative explanations for the lack of success of these MMPIs including the recent finding that some MMPs may play a paradoxical protective role in tumour progression. These observations together with the identification of novel functions for MMPs in early stages of cancer have made necessary a reformulation of MMP inhibition strategies. A better understanding of the functional complexity of this proteolytic system and global approaches to identify the relevant MMPs which must be targeted in each individual cancer patient, will be necessary to clarify whether MMP inhibition may be part of future therapies against cancer. PMID- 15349817 TI - Cytoskeletal mechanisms responsible for invasive migration of neoplastic cells. AB - Cytoskeletal reorganizations, especially alterations of contractile tension generated by the actin-myosin cortex, are of central importance in the development of the phenotype of morphologically transformed neoplastic cells with invasive behavior. These reorganizations can be regarded as genetically determined aberrations of the physiological reactions of normal cells which are responsible for their ability to undergo exploratory migrations, including epithelio-mesenchymal transformations, invasion of matrix by epithelial tubules etc. It is suggested that these physiological and neoplastic transformations are based on Rho-dependent alterations in contractility. A decrease or an increase in contractility may result in the development of distinct types of invasive phenotypes. These contractility-dependent phenotype alterations may be modified by alterations in the expression of other genes, especially of those coding for components of adhesive structures. PMID- 15349818 TI - Collective cell migration in morphogenesis and cancer. AB - The movement of cells that maintain cell-cell junctions yet protrude along or within tissues is an important mechanism for cell positioning in morphogenesis, tissue repair and cancer. Collective cell migration shares similarities but also important differences to individually migrating cells. Coherent groups of cells are arranged and held together by cell-cell adhesion molecules, including cadherins, integrins, ALCAM and NCAM. Integrins of the beta 1 and beta 3 families further provide polarized interactions with the extracellular tissue environment, while matrix-degrading proteases become focalized to substrate contacts to widen tissue space for the advancing cell mass. By generating one functional unit, in contrast to individual cell migration, collective migration provides the active and passive translocation of mobile and non-mobile cells, respectively. This review highlights cellular and molecular principles of collective migration in the context of morphogenic tissue patterning and tumor cell invasion. PMID- 15349819 TI - Invasive growth: a genetic program. AB - Invasive growth is defined as a complex biological program which instructs cells to dissociate, migrate, degrade the surrounding matrix, proliferate and survive. Together, these processes account for tissue morphogenesis, homeostasis and repair, and can be aberrantly implemented for cancer dissemination and metastasis. Individual aspects of this process can be controlled by many cytokines and growth factors. However, coordinated regulation of invasive growth as a whole is specifically accomplished by Hepatocyte Growth Factor, a soluble factor which acts through the tyrosine kinase receptor Met. Here we discuss the different biological facets of invasive growth and analyze the intracellular signals which lead to its execution. PMID- 15349820 TI - From here to there; a life based on migration. An interview with Isaiah J. Fidler. PMID- 15349821 TI - N-cadherin in the spotlight of cell-cell adhesion, differentiation, embryogenesis, invasion and signalling. AB - Cell migration is a process which is essential during embryonic development, throughout adult life and in some pathological conditions. Cadherins, and more specifically the neural cell adhesion molecule N-cadherin, play an important role in migration. In embryogenesis, N-cadherin is the key molecule during gastrulation and neural crest development. N-cadherin mediated contacts activate several pathways like Rho GTPases and function in tyrosine kinase signalling (for example via the fibroblast growth factor receptor). In cancer, cadherins control the balance between suppression and promotion of invasion. E-cadherin functions as an invasion suppressor and is downregulated in most carcinomas, while N cadherin, as an invasion promoter, is frequently upregulated. Expression of N cadherin in epithelial cells induces changes in morphology to a fibroblastic phenotype, rendering the cells more motile and invasive. However in some cancers, like osteosarcoma, N-cadherin may behave as a tumour suppressor. N-cadherin can have multiple functions: promoting adhesion or induction of migration dependent on the cellular context. PMID- 15349822 TI - The Wnt connection to tumorigenesis. AB - Wnt signaling has been identified as one of the key signaling pathways in cancer, regulating cell growth, motility and differentiation. Because of its widespread activation in diverse human tumor diseases, the Wnt pathway has gained considerable and growing interest in tumor research over recent years. Evidence that altered Wnt signaling is important for human tumor development came from three major findings: (i) the tumor suppressor adenomatous polyposis coli (APC) binds to the Wnt pathway component beta-catenin and is involved in its degradation, (ii) mutations of APC in colon tumors lead to stabilization of the beta-catenin protein and (iii) tumor-associated mutations of beta-catenin in colorectal cancer as well as in other tumor types lead to its stabilisation, qualifying beta-catenin as a proto-oncogene. Here we will describe the biochemical interactions which shape the Wnt pathway and focus on its role in tumorigenesis. PMID- 15349823 TI - The chemokine network in cancer--much more than directing cell movement. AB - Cytokine and chemokine gradients are central to the directed movement of cells in both homeostatic and pathological processes. Most cancers have a complex chemokine network which can influence immune responses to the tumor, direct the extent and cellular composition of the leukocyte infiltrate and also play a role in angiogenesis. Tumor cells can also hijack the chemokine system and gain expression of certain chemokine receptors and respond to specific chemokine gradients. Chemokine receptor expression and activation on malignant cells may be central to the growth, survival and migration of cancer cells from the primary tumor. Chemokine receptors, both CC and CXC have been detected on malignant cells and the relevant ligands are sometimes expressed at the tumor site and at sites of tumor spread, suggesting a role for the chemokine family in malignant growth and metastasis. PMID- 15349824 TI - Brain tumour development and invasion. An interview with Ole Didrik Laerum. PMID- 15349825 TI - The stroma reaction myofibroblast: a key player in the control of tumor cell behavior. AB - The cooperation between epithelial and mesenchymal cells is essential for embryonic development and probably plays an important role in pathological phenomena such as wound healing and tumor progression. It is well known that many epithelial tumors are characterized by the local accumulation of connective tissue cells and extracellular material; this phenomenon has been called the stroma reaction. One of the cellular components of the stroma reaction is the myofibroblast, a modulated fibroblast which has acquired the capacity to neoexpress alpha-smooth muscle actin, the actin isoform typical of vascular smooth muscle cells, and to synthesize important amounts of collagen and other extracellular matrix components. It is now well accepted that the myofibroblast is a key cell for the connective tissue remodeling which takes place during wound healing and fibrosis development. Myofibroblasts are capable of remodeling connective tissue but also interact with epithelial cells and other connective tissue cells and may thus control such phenomena as tumor invasion and angiogenesis. In this review we discuss the mechanisms of myofibroblast evolution during fibrotic and malignant conditions and the interaction of myofibroblasts with other cells in order to control tumor progression. On this basis we suggest that the myofibroblast may represent a new important target of antitumor therapy. PMID- 15349826 TI - The countercurrent principle in invasion and metastasis of cancer cells. Recent insights on the roles of chemokines. AB - Chemokine production by cancer cells constitutes a duality. Leukocyte recruitment under the pressure of chemokines may be beneficial for the host or for the tumor. Here, the emphasis will be on the detrimental effects of chemokines in tumor biology. A decade ago, the countercurrent principle of tumor-derived chemokine and peritumoral protease production was formulated to explain chemokine expression as a selective advantage for specific tumors and as a phenotype of invasive and metastasizing cancer cells. Chemoattracted leukocytes may provide trophic factors and produce invasion and metastasis-promoting proteinases. On the basis of the consensus sequence glutamic acid-leucine-arginine (ELR) preceding the canonical cysteine-any amino acid-cysteine (CXC), ELR-positive CXC chemokines, such as interleukin-8 and granulocyte chemotactic protein-2, are angiogenic and thus instruct the host to feed the tumor and bring the vessels into closer contact with the tumor cells. These mechanisms may enhance lymphogenic and hematogenic metastasis. Recent research and proofs of this countercurrent concept are here reviewed and compared. In addition, we discuss how alterations in chemokine ligand and receptor expression profiles may contribute to tumor growth, invasion, metastasis and immune evasion. These comparisons imply practical consequences for future cancer diagnosis and therapy. The implications include methods to diminish metastasis by inhibiting angiogenic CXC chemokine ligands and receptors, therapeutic combinations of chemokine overexpression with antigenic stimuli and co-treatment with angiostatic chemokines and tumor antigens. PMID- 15349827 TI - The migration and differentiation of a chemist entangled in developmental and cancer biology. An interview with Jean-Paul Thiery. PMID- 15349828 TI - Primordial germ cell migration. AB - Mutational and antisense screens in Drosophila and zebrafish, and transcriptional profiling and time-lapse analysis in the mouse, have contributed greatly to our understanding of PGC development. In all three systems, the behavior of PGCs is controlled by growth factors which signal through G-protein coupled receptors and/or tyrosine kinase receptors. Additionally, regulated cell-cell and cell substrate adhesion is important for PGC motility. Finally, localized growth factors may control PGC survival and consequently PGC position. Chemotaxis, regulated adhesion and cell survival are important for multiple migration processes which occur during development and disease. PGC migration shares these features. PMID- 15349829 TI - Germinal tumor invasion and the role of the testicular stroma. AB - Testicular germ cell tumors (TGCTs) are the most frequent neoplasia among young people and their incidence has grown very quickly during recent decades in North America and Europe. Many studies have been carried out in order to elucidate the factors involved in the appearance and progression of these tumors. Little is known about the role of cancer cell-stroma crosstalk in TGCT invasive processes. Here, we review several factors which may be implicated in germ cell tumor progression, such as matrix metalloproteinases, insulin-like growth factor, transforming growth factor beta, the cadherin/catenin complex and integrins. Paradoxically, some of these molecules are also involved in the regulation of normal testicular function. Finally, we discuss prospects for future research on the role of the stroma in the progression and differentiation of male germ cell tumors. PMID- 15349830 TI - Tumor invasion and metastasis: getting more basic to come closer to the patient. An interview with Lance A. Liotta. PMID- 15349831 TI - The "chemoinvasion assay": a tool to study tumor and endothelial cell invasion of basement membranes. AB - Several genetic and epigenetic factors, both in the cell and in the host, contribute to the progression of tumors towards metastases. The escape of cancer cells from a primary, localized tumor to distant organs transforms a relatively curable pathology to an almost untreatable one. Metastatic lesions are often resistant to cancer therapy because of the progressive phenotypic changes that they have undergone. In this article we will give a bird's eye view on the features of metastatic cells and potential therapeutic targets. In particular, the invasion of basement membranes represents a fundamental step for cell dispersion. Over seventeen years ago we established the Matrigel "chemoinvasion" assay, a useful tool for studying the mechanisms involved in tumor and endothelial cell invasion of basement membranes and for the screening of anti invasive agents. We will describe the assay and review some of the major results it enabled to obtain. PMID- 15349832 TI - Novel technologies and recent advances in metastasis research. AB - In this review we have attempted to summarize some of the recent developments in using novel technologies to unravel the molecular mechanisms of tumor progression, in particular the formation of tumor metastasis. In order to push forward the frontiers in cancer research, it is obvious that several fields have to be further developed and interconnected: (1) clinical, epidemiological and pathological studies which mainly use innovative technologies, including microarray technology and nanotechnology to determine as many parameters as possible, (2) the development of improved and suitable bioassays and better animal models and (3) the use of novel computation and bioinformatics methods to sample and integrate the exponentially growing sets of data coming from such investigations. Fashionable as scientists are, this new endeavor may be called systems biology. PMID- 15349833 TI - Parallels in invasion and angiogenesis provide pivotal points for therapeutic intervention. AB - Biological evolution is economical and successful fundamental processes are frequently recapitulated. There are remarkable similarities in the molecular mechanisms which enable tumour cells to invade into surrounding tissues and activated endothelial cells to generate new capillaries, which facilitate the growth and dissemination of cancer. Indeed these pathological processes are themselves based upon key vertebrate developmental processes, and in some cases parallel strategies used by microorganisms to colonise their hosts' tissues. The aim of this review is to explore these parallels in more detail and indicate possible pivotal points for therapeutic intervention. These novel approaches may ultimately optimise the selective targeting of processes involved in tumour invasion and angiogenesis, while sparing normal adult proliferating tissues. Strategies include inhibition of oncogenic pathways in tumour cells which not only stimulate tumour cell growth and invasion, but also initiate neoangiogenesis by upregulation of angiogenic cytokines. Secondly, downstream signalling pathways, transcriptional regulation and effectors common to both processes, and finally points of interaction/cross-talk between tumour cells and endothelial cells which are necessary to enable invasion and angiogenesis to proceed. PMID- 15349834 TI - Pain control by ionizing radiation of bone metastasis. AB - Pain treatment due to cancer is a large fraction of the care in a radiotherapy department. While radiation treatment is very effective in reducing pain, the pathophysiology of bone metastases remains very complex. Reducing the number of tumour cells by radiation will reduce the pressure in bone marrow, but the very rapid response to radiation treatment seen in some patients is probably related to the presence of highly radiosensitive inflammatory cells. In this review we give an overview of the pathophysiological mechanisms which lead to pain associated with bone metastasis and the impact of radiation treatment and other treatments on this mechanism. PMID- 15349835 TI - Expression of peroxiredoxins in bovine oocytes and embryos produced in vitro. AB - Peroxiredoxins (PRDXs) form a family of peroxidases involved in antioxidant protection and cell signaling. Due to their peroxide reductase activity, these enzymes might be involved in fine-tuning peroxide levels in embryos during in vitro production. In this study, RT-PCR was used to examine the expression of the six PRDX isoforms (PRDX1 to PRDX6) in bovine oocytes and embryos. PRDXs were detected in oocytes both before and after in vitro maturation. Besides, PRDX6 was up-regulated after maturation. Single embryos were analyzed from the two-cell to the blastocyst stages. PRDX1 and PRDX5 transcripts were detected throughout development. PRDX2, PRDX3, and PRDX6 were not expressed around the 9- to 16-cell stage. PRDX4 transcripts were weakly detected in pools of embryos from the 9- to 16-cell stage onwards. In situ immunodetection of PRDX5, which was previously reported to exhibit the widest subcellular distribution among PRDXs in adult mammalian cells, showed a mitochondrial distribution pattern in the bovine embryo. Finally, the potential modulation by oxidative stress of PRDX expression around the major embryonic genome activation was evaluated by culturing embryos under 20% O2 instead of 5%. No significant difference in the pattern of PRDX expression was observed under 20% O2. In conclusion, our data show for the first time that PRDXs are expressed in mammalian oocytes and early embryos. Moreover, the bovine transcripts exhibit various patterns of expression that might be related to the potential role of PRDXs in oocyte maturation and embryo development. PMID- 15349836 TI - Uterine secretion of ISP1 & 2 tryptases is regulated by progesterone and estrogen during pregnancy and the endometrial cycle. AB - We have described two novel implantation serine proteinase (ISP) genes that are expressed during the implantation period. The ISP1 gene may encode the embryo derived enzyme strypsin, which is necessary for blastocyst hatching in vitro and the initiation of invasion. The ISP2 gene, which encodes a related tryptase, is expressed in endometrial glands and is regulated by progesterone during the peri implantation period. Based on similarities between ISP2 gene expression and that of a progesterone-regulated lumenal serine proteinase activity associated with lysis of the zona pellucida, we have suggested that the strypsin related protein, ISP2, may encode a zona lysin proteinase. Recently strypsin has also been found within uterine fluid, suggesting a second potential role in hatching. Consistently, we have discovered that ISP1 is also expressed in the uterine secretory gland at the time of hatching. In this study we demonstrate that both ISP1 and ISP2 are secreted together into the uterine lumen at peri-implantation, and that the appearance of ISP protein is regulated positively at the transcriptional level by progesterone and negatively at the posttranscriptional level by estrogen. This negative regulation by estrogen may be overridden in pregnancy as ISP protein expression is restored during oil-induced decidualization. ISP1 and ISP2 proteins are also expressed in proestrous suggesting additional roles in the endometrial cycle. PMID- 15349837 TI - Transcription factor Sox-2 inhibits co-activator stimulated transcription. AB - Previous studies have shown that transcription of the fibroblast growth factor-4 (FGF-4) gene by early embryonic cells is dependent upon a powerful distal enhancer located 3 kb downstream of the transcription start site within the untranslated region of the last exon. The transcription factors Sox-2 and Oct-3 cooperatively bind to critical cis-regulatory elements within the enhancer to synergistically activate transcription. Moreover, the co-activator p300 can mediate the synergistic activity of Sox-2 and Oct-3, and p300 associates with the FGF-4 enhancer in vivo. Embryonal carcinoma (EC) cells have been used extensively as a model system to study the regulation of the FGF-4 gene during early development. Recently, it has been suggested that suboptimal levels of Sox-2 expression in F9 EC cells limit the transcription of the FGF-4 gene. The studies presented in this report argue that Sox-2 levels are not limiting in F9 EC cells. Moreover, overexpression of Sox-2 in F9 EC cells decreases FGF-4 promoter activity. In addition, overexpression of Sox-2 in these cells inhibits activation by the co-activators p300, CBP, and OCA-B in a manner that requires the transactivation domain of Sox-2. These findings suggest that Sox-2 levels in F9 EC cells are regulated carefully to avoid interference with the transcription of critical genes. PMID- 15349838 TI - Nonviral gene transfer to surface skin of mid-gestational murine embryos by intraamniotic injection and subsequent electroporation. AB - The surface epithelium of mid-gestational murine embryos is thought to be an attractive target for gene therapy in vivo, due to its visibility and accessibility from the external surface of the maternal uterus. Almost all studies of in utero gene transfer have adopted viral vectors for infection of fetal epithelium, and depended on intraamniotic introduction and simple incubation of vectors, leading to only infection of the surface layer (periderm) of fetal skin. Here we report a simple and convenient method of gene transfer of plasmid DNA into the deeper portion of surface skin of murine mid-gestational fetus. One to two microlitres of a solution containing a lacZ expression plasmid (0.5-1 microg) and trypan blue (0.05%) were placed onto the surface of a fetus (E 14.5) near the eye by a micropipette attached to a mouthpiece. This fetus was immediately electroporated by placing it between tweezer-type electrodes attached to a square-pulse generator. At 1 and 4 days after gene transfer, fetuses were subjected to histochemical staining for lacZ activity in the presence of X-Gal, a substrate for lacZ. Focal reactions were observed in the skin epidermal layers including periderm and basal layer 1 day after DNA introduction. However, lacZ positive cells were limited to a skin surface layer, the stratum corneum, in the samples obtained 4 days after gene transfer. Similar observation was also made in the transgenic fetuses (carrying a lacZ gene placed immediately downstream of the loxP-flanked sequence) injected with Cre expression vector. These findings suggest rapid movement of fetal epidermal cells toward the surface during late developmental stages. This local gene transfer approach appears to be effective as a method for skin-targeted gene transfer, enabling study of the role of genes of interest and tracing of cell lineage during fetal skin development. PMID- 15349839 TI - Birth of calves expressing the enhanced green fluorescent protein after transfer of fresh or vitrified/thawed blastocysts produced by somatic cell nuclear transfer. AB - The present study examined effects of genetic manipulation and serum starvation on in vitro developmental potential of bovine somatic cell nuclear transfer (SCNT) embryos and vitrification on in vivo developmental competence of transgenic SCNT blastocysts. Fetal oviduct epithelial cells (FOECs) were isolated from the oviduct of a Day 147 bovine fetus and transfected with a plasmid (pCE EGFP-IRES-NEO) containing the enhanced green fluorescent protein (EGFP) and neomycin-resistant (Neor) genes. There were no significant differences (P > 0.05) in cleavage rates or development rates to the blastocyst stage for SCNT embryos derived from FOECs (72.5 and 47.8%, respectively) or transfected FOECs (TFOECs, 73.8 and 47.7%, respectively); nor from serum-fed (73.6 and 47.2%, respectively) or serum-starved (72.7 and 48.3%, respectively) cells. Seventeen of Day 7 GFP embryos (eight fresh blastocysts and nine vitrified/thawed blastocysts ) were transferred to recipients with one embryo per recipient. Two (25%) recipients were confirmed pregnant at Day 60 in fresh blastocysts group, and three recipients (33%) were confirmed pregnant at Day 60 in vitrified/thawed blastocysts group. Two healthy calves (25%) were obtained from fresh blastocysts and one (11%) from vitrified/thawed blastocysts. Microsatellite analysis confirmed that the three clones were genetically identical to the donor cells. Moreover, PCR and Southern blot demonstrated integration of transgene in genomic DNA of all three cloned calves. Expression of GFP in skin biopsies isolated from transgenic cloned calves and fibroblasts derived from the skin biopsies revealed the activity of EGFP gene, and G418 resistance in vitro of these fibroblasts confirmed the activity of Neor gene. Our results show that genetic manipulation and serum starvation of donor cells (FOECs) do not affect in vitro developmental competence of bovine SCNT embryos, and vitrified transgenic SCNT blastocysts can develop to term successfully. PMID- 15349840 TI - Effect of activation time on the nuclear remodeling and in vitro development of nuclear transfer embryos derived from bovine somatic cells. AB - This study was conducted to investigate the effect of recipient activation time on the chromatin structure and development of bovine nuclear transfer embryos. Serum-starved skin cells were electrofused to enucleated oocytes, activated 1-5 hr after fusion, and cultured in vitro. Some fused eggs were fixed at each time point after fusion without activation, or 3 or 7 hr after activation. Some nocodazole treated zygotes were fixed to analyze their chromosome constitutions. The proportion of eggs with a morphologically normal premature chromosome condensation (PCC) state increased 1-2 hr after fusion. Whereas eggs with elongated chromosome plate increased as activation time was prolonged to 3 hr, and 5 hr after fusion, 58.1% of eggs showed more than two scattered chromosome sets. The proportion of eggs with a single chromatin mass (40.6-56.7%) significantly increased when eggs were activated within 2.5 hr after fusion (P < 0.05). Only 23.3% of reconstituted embryos activated 5 hr after fusion formed one pronucleus-like structure (PN), whereas, 64.5-78.3% of embryos activated 1-2.5 hr after fusion formed one PN. The proportion of embryos with normal chromosome constitutions decreased as activation time was prolonged. Development rates to the blastocyst stage were higher in eggs activated within 2 hr after fusion (17.3 21.7%) compared to those of others (0-8.6%, P < 0.05). The result of the present study suggests that activation time can affect the chromatin structure and in vitro development of bovine nuclear transfer embryos. PMID- 15349841 TI - Blastocysts produced by nuclear transfer between chicken blastodermal cells and rabbit oocytes. AB - Interspecies nuclear transfer (INT) has been used as an invaluable tool for studying nucleus-cytoplasm interactions; and it may also be a method for rescuing endangered species whose oocytes are difficult to obtain. In the present study, we investigated interaction of the chicken genome with the rabbit oocyte cytoplasm. When chicken blastodermal cells were transferred into the perivitelline space of rabbit oocytes, 79.3% of the couplets were fused and 9.7% of the fused embryos developed to the blastocyst stage. Both M199 and SOF medium were used for culturing chicken-rabbit cloned embryos; embryo development was arrested at the 8-cell stage obtained in SOF medium, while the rates of morulae and blastocysts were 12.1 and 9.7%, respectively, in M199 medium. Polymerase chain reaction (PCR) amplification of nuclear DNA and karyotype analyses confirmed that genetic material of morulae and blastocysts was derived from the chicken donor cells. Analysis mitochondrial constitution of the chicken-rabbit cloned embryos found that mitochondria, from both donor cells and enucleated oocytes, co-existed. Our results suggest that: (1) chicken genome can coordinate with rabbit oocyte cytoplasm in early embryo development; (2) there may be an 8- to 16-cell stage block for the chicken-rabbit cloned embryos when cultured in vitro; (3) mitochondrial DNA from the chicken donor cells was not eliminated until the blastocyst stage in the chicken-rabbit cloned embryos; (4) factors existing in ooplasm for somatic nucleus reprogramming may be highly conservative. PMID- 15349842 TI - Expression of HSP70/HSC70 in swine blastocysts: effects of oxidative and thermal stress. AB - Stress proteins are induced in response to a wide range of biological and physicochemical stresses; HSC70 (constitutive) and HSP70 (inducible) belong to the HSP70 stress protein family. The present study aimed at investigating whether environmental stress, particularly thermal and oxidative stress, is involved in modulating HSP70/HSC70 expression in in vitro porcine embryos from two/four cell stage to blastocyst. For oxidative stress, embryos were cultured at 38.5 degrees C under a 5% O2-5% CO2 atmosphere or 5% CO2 in air (approximately 20% O2); for thermal stress, embryos were cultured at 38.5 degrees C under 5% O2, 5% CO2, and exposed to heat shock (1 hr at 42 degrees C). At the end of culture, embryos were analysed by Western blotting, using specific antibodies discriminating HSP70 from HSC70. Embryos cultured under 20% O2 showed HSC70 levels significantly higher (P < 0.005) than embryos cultured under 5% O2, while heat shocked embryos presented HSP70 levels higher (P < 0.01) than control group. In addition, the developmental rate of embryos was negatively affected by the higher oxygen tension (P < 0.05). Our data indicate that porcine embryos express both HSP70 and HSC70 and could differentially respond to both oxidative and heat stress by up-regulating HSC70 and HSP70, respectively. PMID- 15349843 TI - Regulation of cleavage by protein kinase C in Chaetopterus. AB - We report that protein kinase C (PKC) plays a regulatory role in early cleavage in Chaetopterus eggs. Using Western blotting, we assayed the expression patterns of conventional PKCs (cPKC), novel PKCs (nPKC), and atypical PKCs (aPKC). During early development after fertilization, PKC protein levels varied independently by isoform. PKC protein expression during differentiation, without cleavage and after parthenogenetic activation, was very similar to that during normal development indicating that PKC gene expression does not require cellularization. Since PKC has been shown to regulate meiosis in this organism, we also assayed the membrane association of these isoforms as an indicator of their activation during meiosis and early cleavage. PKC-gamma transiently associated with membranes and therefore became activated before meiotic division and cleavage, whereas PKC-alpha and -beta transiently dissociated from membranes and therefore became inactivated at these times. Inhibition of these PKC isoforms by bisindolylmaleimide I had no effect on cleavage or early development to the trochophore larva, indicating that PKC-gamma activation is not essential for cleavage or early development. However, their persistent activation by thymeleatoxin blocked cleavage. The results indicate that the dissociation of PKC alpha and/or -beta from the membrane fraction, and therefore their inactivation, is essential for normal cleavage. Elevated PKC activity is essential for nuclear envelope breakdown and spindle formation at meiosis I. By contrast, down regulation of this activity is essential for cleavage after fertilization. PMID- 15349844 TI - Identification of Ca2+-dependent calmodulin-binding proteins in rat spermatogenic cells as complexes of the heat-shock proteins. AB - Ca2+-calmodulin (CaM)-binding proteins in rat testes were characterized by assays for CaM-binding activity using the CaM-overlay method on transblots of electrophoresed gels and purification by gel-filtration, ion exchange, and adsorption chromatographies. A major CaM-binding protein complex (CaMBP) was identified and found to be comprised of three proteins with molecular masses 110, 100, and 70 kDa. Amino acid sequence analyses of lysylendopeptidase digests from these proteins indicated that all of the constituents of CaMBP are very similar to the members of the heat-shock protein family, i.e., the 110-kDa protein is similar to the APG-2/94 kDa rat ischemia-responsive protein, the 100-kDa protein is similar to the rat counterpart of the mouse APG-1/94 kDa osmotic stress protein, and the 70-kDa protein is similar to the rat testis-specific major heat shock protein (HSP70). Immunohistochemistry using anti-CaMBP and anti-CaM antibodies demonstrated that CaMBP was co-localized with CaM in the cytoplasm of pachytene spermatocytes and nuclei of round spermatids. In addition, CaMBP, but not CaM, was localized at a high level in the residual bodies of elongated spermatids. The possible relevance of CaMBP to regulation of cell cycle progression and spermatogenesis is discussed in this paper. PMID- 15349845 TI - Macaque sperm release ESP13.2 and PSP94 during capacitation: the absence of ESP13.2 is linked to sperm-zona recognition and binding. AB - ESP13.2 coats the entire surface of macaque sperm and remains until sperm become capacitated (Yudin et al., 2003: Biol Reprod 69: 1118-1128). Capacitation of macaque sperm is synchronized by treatment with dibutyrl cAMP (dbcAMP) and caffeine. ESP13.2 and PSP94 constituted approximately 95% of the proteins released from the sperm surface following treatment with caffeine + dbcAMP. Caffeine and dbcAMP alone induce different patterns of ESP13.2 release. As determined by ELISAs of supernatants and immuno-fluorescent labeling of sperm heads, caffeine alone and caffeine + dbcAMP induced comparable release of ESP13.2, while dbcAMP-treated sperm did not differ from controls. Sperm treated with caffeine + dbcAMP showed a reduction of ESP13.2 from the entire surface, while caffeine treatment alone induced removal of ESP13.2 from the sperm head and midpiece. As confirmed with immunofluorescence, ESP13.2 could be added back to the surfaces of sperm that had been previously exposed to caffeine. Treatment with caffeine significantly increased the number of sperm that bound tightly to the zona pellucida as compared with controls (42 +/- 9 and 13 +/- 3 sperm/zona, respectively; P < or = 0.01). This increase in binding was inhibited by "adding back" ESP13.2 to the sperm surface (12.8 +/- 3; P < or = 0.01). Alexa-conjugated anti-ESP13.2 Ig labeling of live sperm showed that only sperm lacking ESP13.2 over the head were capable of tight binding to the zona. Our results suggest that ESP13.2 masks zona pellucida ligands on the sperm surface and its release, as part of capacitation, is required for sperm-zona interaction. PMID- 15349846 TI - Effect of macromolecule supplementation during in vitro maturation of goat oocytes on developmental potential. AB - In vitro maturation (IVM) of goat oocytes with serum-supplemented media results in oocytes with reduced developmental potential. The objective of this study was to develop a defined medium for IVM of goat oocytes that better supports subsequent embryonic development. Cumulus oocyte complexes (COC) were matured for 18-20 hr in: Experiment (1), tissue culture medium 199 (TCM199) with 10% (v/v) goat serum or modified synthetic oviduct fluid maturation medium (mSOFmat) with 2.5, 8.0, or 20.0 mg/ml bovine serum albumin (BSA); Experiment (2), mSOFmat with 4.0, 8.0, 12.0, or 16.0 mg/ml BSA; or Experiment (3), 1.0 mg/ml polyvinyl alcohol (PVA; control), 4.0 mg/ml BSA, 0.5 mg/ml hyaluronate plus 0.5 mM citrate, or hyaluronate, citrate, and BSA. Mature COC were coincubated for 20-22 hr with 12 15 x 10(6) sperm/ml in modified Brackett and Oliphant (mBO) medium. Embryos were cultured for a total of 7 days in G1/2, and evaluated for cleavage, and blastocyst development, hatching, and total cell numbers. In the first experiment, more (P < 0.05) blastocysts developed per cleaved embryo following maturation in mSOFmat with 2.5 or 8.0 mg/ml BSA than with 20.0 mg/ml BSA or TCM199 with 10% goat serum. The various concentrations of BSA used in the second experiment did not affect (P > 0.05) any of the developmental endpoints examined. In the third experiment, developmental potential of oocytes matured with PVA or hyaluronate with citrate was not different (P > 0.05) from oocytes matured in the presence of BSA. These results demonstrate that developmentally competent goat oocytes can be matured under defined conditions. PMID- 15349847 TI - Hormonal control of somatic cell oocyte interactions during ovarian follicle development. AB - In the mammalian ovarian follicle, paracrine signaling between the oocyte and somatic granulosa cells is bidirectional but the structural basis and physiological regulations of communication between gametic and somatic compartments remain unknown. The present experiments were designed to test the hypothesis that follicle stimulating hormone (FSH) regulates the ability of granulosa cells to make connections with the oocyte. We show that in prepubertal unprimed mice and mice carrying a targeted deletion of the FSHbeta subunit gene, granulosa cells exhibit orientation towards the oocyte manifest by the elaboration of transzonal projections (TZPs) and "apical" centrosome positioning at sites of granulosa-zona contact. In vivo FSH treatment results in a retraction of TZPs. Coincident with TZP retraction induced by FSH are changes in oocyte transcriptional activity and meiotic competence, which suggests one means by which the oocyte-granulosa cell dialogue may be modulated during development of ovarian follicles. PMID- 15349848 TI - Molecular modification of Penaeus monodon sperm in female thelycum and its consequent responses. AB - Using Penaeus monodon as the model, we demonstrated the molecular changes and the mechanism of thelycal-dependent sperm modification resulting in an enhanced acrosome reaction (AR) response. Attention was paid to the modification of the sperm plasma membrane which was mediated through an adsorption or removal of sperm peripheral and integral membrane proteins as indicated by the different profiles of these proteins in spermatophore (S) and thelycal (T) sperm. In vitro adsorption of Alexa-488 conjugated T proteins onto the entire S-sperm surface confirmed protein transfer in a time-dependent manner. Specific anchoring of 83 and 140 kDa proteins to sperm peripheral proteins as well as 53/55 and 60 kDa proteins to sperm lipids was demonstrated. Apart from membrane modification, a substantial increase in protein tyrosine phosphorylation was shown to be closely associated with T-dependent sperm modification event. The physiological significance of this sperm modification in enhancing sperm AR response, which required at least 3 days of T residence in order for the sperm to gain a complete AR response, was also elucidated. PMID- 15349849 TI - Neurology Outcomes Research Meeting, October 3, 2004, Toronto, Ontario, Canada. Abstracts. PMID- 15349851 TI - 129th Annual Meeting American Neurological Association, October 3-6, 2004, Toronto, Ontario, Canada. Abstracts. PMID- 15349853 TI - 33nd Annual Meeting of the Child Neurology Society, October 13-16, 2004, Ottawa, Canada. Abstracts. PMID- 15349856 TI - Roles of the proteasome in neurodegenerative disease: refining the hypothesis. PMID- 15349857 TI - How many pathways are there to nigral death? PMID- 15349858 TI - Generalized brain and skin proteasome inhibition in Huntington's disease. AB - Mutated intracellular huntingtin is widely expressed in tissues of Huntington's disease (HD) patients. Intraneuronal nuclear protein aggregates of mutant huntingtin are present in HD brains, suggesting a dysfunction of the ubiquitin proteasome system (UPS). Because many cells and tissues can cope with the abnormal gene effects while others dysfunction and die, we determined gene induced effects and considered the hypothesis that the gene causes multiple intracellular problems, but severe pathology is seen only in selected brain regions. In this study, we found inhibition of UPS function in both early (0-1, with no or little neuronal loss) and late (3-4, with more severe neuronal loss) stage HD patients' cerebellum, cortex, substantia nigra and caudate-putamen brain regions. Late HD stage increases in ubiquitin levels were unique to caudate putamen. HD patients' skin fibroblasts also had UPS inhibition similar to brain despite increases in proteasome beta-subunit expression. Gene delivery and expression of proteasome activator PA28 increased UPS function in normal but not HD fibroblasts. These generalized UPS problems are associated with severe neuronal pathology only when coupled with decreases in brain-derived neurotrophic factor levels, mitochondrial complex II/III activity, and increases of ubiquitin levels particularly as seen in the caudate-putamen of HD patients. PMID- 15349859 TI - The gene responsible for PARK6 Parkinson's disease, PINK1, does not influence common forms of parkinsonism. AB - Mutations in the PINK1 gene (PARK6), a putative serine-threonine kinase, cause autosomal recessive Parkinson's disease. PINK1 functions as a protein kinase and confers protective effects in the mitochondria, where it is primarily located. We assessed in a population of European ancestry whether common genetic variation in this novel gene influences nonmendelian forms of Parkinson's disease. We defined the linkage disequilibrium structure of PINK1 and used this to identify a set of tagging single nucleotide polymorphisms that we estimate will efficiently represent all of the common DNA variation in the entire gene. Genotyping these tags in a set of 576 Parkinson's disease patients and 514 controls did not demonstrate a case-control partition for allele or for haplotype and thus provides evidence against the existence of a common functional variants in PINK1 that has a strong influence on PD risk. PMID- 15349860 TI - PINK1 mutations are associated with sporadic early-onset parkinsonism. AB - We have recently reported homozygous mutations in the PINK1 gene in three consanguineous families with early-onset parkinsonism (EOP) linked to the PARK6 locus. To further evaluate the pathogenic role of PINK1 in EOP and to draw genotype-phenotype correlates, we performed PINK1 mutation analysis in a cohort of Italian EOP patients, mostly sporadic, with onset younger than 50 years of age. Seven of 100 patients carried missense mutations in PINK1. Two patients had two PINK1 mutations, whereas in five patients only one mutation was identified. Age at onset was in the fourth-fifth decade (range, 37-47 years). The clinical picture was characterized by a typical parkinsonian phenotype with asymmetric onset and rare occurrence of atypical features. Slow progression and excellent response to levodopa were observed in all subject. Two of 200 healthy control individuals also carried one heterozygous missense mutation. The identification of a higher number of patients (5%) than controls (1%) carrying a single heterozygous mutation, along with previous positron emission tomography studies demonstrating a preclinical nigrostriatal dysfunction in PARK6 carriers, supports the hypothesis that haploinsufficiency of PINK1, as well as of other EOP genes, may represent a susceptibility factor toward parkinsonism. However, the pathogenetic significance of heterozygous PINK1 mutations still remains to be clarified. PMID- 15349861 TI - Impaired cell adhesion and apoptosis in a novel CLN9 Batten disease variant. AB - We describe the ninth variant of neuronal ceroid lipofuscinosis (NCL) or Batten disease, due to defects in a putative new gene, CLN9. We therefore refer to the new variant as CLN9-deficient. Two Serbian sisters and two German brothers are described. Their clinical history is characteristic for juvenile NCL. They show similar gene expression patterns. The existence of this variant is supported by the presence of curvilinear inclusions, fingerprint profiles, and granular osmiophilic deposits in neurons, lymphocytes, and conjunctival cells. Enzyme screening and sequencing of the coding regions of other NCL genes was negative. CLN9-deficient cells have a distinctive phenotype. They have rounded cell bodies, have prominent nucleoli, attach poorly to the culture dish, and are sensitive to apoptosis but have increased growth rates. Gene expression of proteins involved in cell adhesion and apoptosis is altered in these cells. Sphingolipid metabolism is also perturbed. They have decreased levels of ceramide, sphingomyelin, lactosylceramide, ceramide trihexoside, and globoside and increased activity of serine palmitoyl transferase. PMID- 15349862 TI - Anticancer agents are potent neurotoxins in vitro and in vivo. AB - Neurotoxicity of anticancer agents complicates treatment of children with cancer. We investigated neurotoxic effects of common cytotoxic drugs in neuronal cultures and in the developing rat brain. When neurons were exposed to cisplatin (5-100 microM), cyclophosphamide (5-100 microM), methotrexate (5-100 microM), vinblastin (0.1-1 microM), or thiotepa (5-100 microM), a concentration-dependent neurotoxic effect was observed. Neurotoxicity was potentiated by nontoxic glutamate concentrations. The N-methyl-D-aspartate receptor antagonist MK 801 (10 microM), the AMPA receptor antagonists GYKI 52466 (10 microM) and NBQX (10 microM), and the pancaspase inhibitor Ac-DEVD-CHO (1 nM) ameliorated neurotoxicity of cytotoxic drugs. To investigate neurotoxicity in vivo, we administered to 7-day old rats the following: cisplatin (5-15 mg/kg i.p.), cyclophosphamide (200-600 mg/kg i.p.), thiotepa (15-45 mg/kg), or ifosfamide (100-500 mg/kg) and their brains were analyzed at 4 to 24 hours. Cytotoxic drugs produced widespread lesions within cortex, thalamus, hippocampal dentate gyrus, and caudate nucleus in a dose-dependent fashion. Early histological analysis demonstrated dendritic swelling and relative preservation of axonal terminals, which are morphological features indicating excitotoxicity. After longer survival periods, degenerating neurons displayed morphological features consistent with active cell death. These results demonstrate that anticancer drugs are potent neurotoxins in vitro and in vivo; they activate excitotoxic mechanisms but also trigger active neuronal death. PMID- 15349863 TI - Genotype determines phenotype in experimental Lyme borreliosis. AB - Borrelia burgdorferi sensu lato, the causative organism of Lyme borreliosis, is a heterogeneous group of spirochetes, consisting of at least three pathogenic species. To test the hypothesis that the genetic heterogeneity is the reason for the clinical differences, we investigated whether the experimental disease induced by European isolates is different from that induced by American isolates. Two American isolates of species B. burgdorferi sensu stricto were compared with three European isolates, two of species B. garinii, and one of species B. afzelii. The patterns of infection, immunity, and inflammation induced by the different species was distinctive. Inflammatory cells and levels of antibody in B. garinii- and B. afzelii-infected animals were lower than in B. burgdorferi s.s.-infected animals, whereas levels of spirochetal infection in the skin and nervous system were higher in the former group of animals. These data demonstrate that B. burgdorferi s.s. strains are more infective and inflammatory, whereas B. garinii and B. afzelii strains can survive the adaptive immune response to a greater degree and persist at greater numbers in the skin and nervous system. The results explain to a large extent the disparities between LNB in humans in the United States and Europe. PMID- 15349864 TI - Activation of 5-HT(1B/1D) receptor in the periaqueductal gray inhibits nociception. AB - It is considered that the site of action of the abortive antimigraine compounds acting at serotonin, 5-HT(1B/1D,) receptors (triptans) is the trigeminovascular system. We tested whether there is a non-trigeminal site of action. The 5 HT(1B/1D) agonist, naratriptan, was microinjected into the ventrolateral periaqueductal gray (vlPAG), and activity in the trigeminal nucleus caudalis (TNC) was monitored. Recordings were made from 20 nociceptive neurons in the dorsal horn of the TNC that received convergent input from the dura mater and face. Responses of neurons to dural, facial cutaneous and corneal stimulation were studied before and after injection of naratriptan. Naratriptan decreased the excitability to electrical stimulation of the dura mater as the A-fiber response decreased by 24 +/- 4.1% (p < 0.001) and the C-fiber response decreased by 42 +/- 8.2% (p < 0.001). Spontaneous activity was decreased by 38 +/- 7.5% (p < 0.001). After injection, the mechanical thresholds of the dura mater increased from (n = 14, p < 0.01). Responses to stimulation of the face and cornea were not altered by injection of naratriptan. These results suggest that 5-HT(1B/1D) receptor activation in the vlPAG activates descending pain-modulating pathways that inhibit dural, but not facial and corneal nociceptive input. These findings have implications for the understanding of the action of triptans in migraine and cluster headache, suggesting that brain loci other than the trigeminal nucleus may play a role in the clinical action of triptans. PMID- 15349865 TI - A narcolepsy susceptibility locus maps to a 5 Mb region of chromosome 21q. AB - The genetic basis of human narcolepsy remains poorly understood. Multiplex families with full-blown narcolepsy-cataplexy are rare, whereas families with both narcolepsy-cataplexy and excessive daytime sleepiness without cataplexy are more common. We performed a genomewide linkage analysis in a large French family with four members affected with narcolepsy-cataplexy and 10 others with isolated recurrent naps or lapses into sleep. Only three regions showed logarithm of odds (LOD) scores greater than 1 in two-point linkage analysis (D6S1960, D11S2359, and D21S228). Genotyping additional markers provided support for linkage to 9 markers on chromosome 21 (maximum two-point LOD score, 3.36 at D21S1245). The multipoint linkage analysis using SimWalk2 provided further evidence for linkage to the same region (maximum parametric LOD score, 4.00 at 21GT26K). A single haplotype was shared by all affected individuals and informative crossovers indicated that the elusive gene that confers susceptibility to narcolepsy is likely to be located between markers D21S267 and ABCG1, in a 5.15 Mb region of 21q. PMID- 15349866 TI - New England Medical Center Posterior Circulation registry. AB - Among 407 New England Medical Center Posterior Circulation registry patients, 59% had strokes without transient ischemic attacks (TIAs), 24% had TIAs then strokes, and 16% had only TIAs. Embolism was the commonest stroke mechanism (40% of patients including 24% cardiac origin, 14% intraarterial, 2% cardiac and arterial sources). In 32% large artery occlusive lesions caused hemodynamic brain ischemia. Infarcts most often included the distal posterior circulation territory (rostral brainstem, superior cerebellum and occipital and temporal lobes); the proximal (medulla and posterior inferior cerebellum) and middle (pons and anterior inferior cerebellum) territories were equally involved. Severe occlusive lesions (>50% stenosis) involved more than one large artery in 148 patients; 134 had one artery site involved unilaterally or bilaterally. The commonest occlusive sites were: extracranial vertebral artery (52 patients, 15 bilateral) intracranial vertebral artery (40 patients, 12 bilateral), basilar artery (46 patients). Intraarterial embolism was the commonest mechanism of brain infarction in patients with vertebral artery occlusive disease. Thirty-day mortality was 3.6%. Embolic mechanism, distal territory location, and basilar artery occlusive disease carried the poorest prognosis. The best outcome was in patients who had multiple arterial occlusive sites; they had position-sensitive TIAs during months to years. PMID- 15349867 TI - Clinicopathological correlates in frontotemporal dementia. AB - The term frontotemporal dementia (FTD) encompasses a range of clinical syndromes that are believed not to map reliably onto the spectrum of recognized pathologies. This study reexamines the relationships between clinical and pathological subtypes of FTD in a large series from two centers (n = 61). Clinical subtypes defined were behavioral variant FTD (n = 26), language variants (semantic dementia, n = 9; and progressive nonfluent aphasia, n = 8), and motor variants (corticobasal degeneration, n = 9; and motor neuron disease, n = 9), although most cases presented with a combination of behavioral and language problems. Unexpectedly, some behavioral cases (n = 5) had marked amnesia at presentation. The pathological subtypes were those with tau-immunopositive inclusions (with Pick bodies, n = 20; or without, n = 11), those with ubiquitin immunopositive inclusions (n = 16), and those lacking distinctive histology (n = 14). Behavioral symptoms and semantic dementia were associated with a range of pathologies. In contrast, other clinical phenotypes had relatively uniform underlying pathologies: motor neuron disease predicted ubiquitinated inclusions, parkinsonism and apraxia predicted corticobasal pathology, and nonfluent aphasia predicted Pick bodies. Therefore, the pathological substrate can be predicted in a significant proportion of FTD patients, which has important implications for studies targeting mechanistic treatments. PMID- 15349868 TI - Magnetization transfer ratio and myelin in postmortem multiple sclerosis brain. AB - Several quantitative magnetic resonance (MR) measures are used to investigate multiple sclerosis (MS) in vivo. Precise quantitative investigation of the histopathological correlates of such measures has, to date, been limited. This study investigates the relationship of quantitative measures of myelin content, axonal density, and gliosis with quantitative MR measures in postmortem (PM) MS tissue. MR imaging (MRI) was performed on a 1.5T scanner and T1-relaxation time (T1-RT) and magnetization transfer ratio (MTR) maps were acquired in fresh PM brain of 20 MS subjects. Myelin content, axonal counts, and the extent of gliosis all were quantified using morphometric and digital imaging techniques. MRI and pathological data were in most cases coregistered using stereotactic navigation. Using multiple regression analysis, we detected significant correlations between myelin content (Tr(myelin)) and MTR (r = -0.84, p < 0.001) and myelin content and axonal count (-0.80, p < 0.001); MTR correlated with T1-RT (r = -0.79, p < 0.001). No association was detected between the extent of gliosis and either MR measure. MTR was significantly higher in remyelinated than demyelinated lesions (means: 30.0 [standard deviation, 2.9] vs 23.8 [standard deviation, 4.3], p = 0.008). In conclusion, MTR is affected by myelin content in MS white matter. PMID- 15349869 TI - Enhancing antiretroviral therapy for human immunodeficiency virus cognitive disorders. AB - The benefits of combination antiretroviral therapy (ART) for HIV cognitive disorders vary substantially between individuals. This study evaluated whether cerebrospinal fluid (CSF) drug penetration and CSF virological suppression influence the extent of neuropsychological (NP) improvement during ART. Overall performance on a battery of NP tests administered at baseline and follow-up (median 15 weeks) was computed by using the global deficit score (GDS) methods in 31 cognitively impaired, HIV-infected individuals who began new ART regimens. Virological suppression (attaining undetectable viral load by RT-PCR at follow up) was assessed separately for plasma and CSF. Subjects on regimens containing greater numbers of CSF-penetrating drugs showed significantly greater reduction in CSF viral load. Subjects attaining CSF virological suppression demonstrated greater GDS improvement than those who did not (median GDS change, 0.62 vs 0.23; p = 0.01). A similar trend for plasma did not reach statistical significance (p = 0.053). NP improvement was greater in ART-naive versus treatment-experienced subjects. In a multivariate model (overall p = 0.0008), significant, independent predictors of GDS reduction were CSF HIV RNA suppression, baseline antiretroviral history, and their interaction. Including CSF-penetrating drugs in the ART regimen and monitoring CSF viral load may be indicated for individuals with HIV associated cognitive impairment. PMID- 15349870 TI - Novel PINK1 mutations in early-onset parkinsonism. AB - PINK1 was recently found to be associated with PARK6 as the causative gene. We performed mutation analysis in eight inbred families whose haplotypes link to the PARK6 region. We identified six pathogenic mutations (R246X, H271Q, E417G, L347P, and Q239X/R492X) in six unrelated families. All sites of mutations were novel, suggesting that PINK1 may be the second most common causative gene next to parkin in parkinsonism with the recessive mode of inheritance. PMID- 15349871 TI - Homozygous PINK1 C-terminus mutation causing early-onset parkinsonism. AB - Two homozygous mutations in the PINK1 gene, encoding a mitochondrial putative protein kinase, recently have been identified in families with PARK6-linked, autosomal recessive early-onset parkinsonism (AREP). Here, we describe a novel homozygous mutation (1573_1574 insTTAG) identified in an AREP patient, which causes a frameshift and truncation at the C-terminus of the PINK1 protein, outside the kinase catalytic domain. The clinical phenotype includes early-onset (28 years) parkinsonism, foot dystonia at onset, good levodopa response, slow progression, early levodopa-induced dyskinesias, and sleep benefit, thereby resembling closely parkin-related disease. These findings confirm that recessive mutations in PINK1 cause early-onset parkinsonism and expand the associated clinical phenotype. PMID- 15349872 TI - Frequency of catamenial seizure exacerbation in women with localization-related epilepsy. AB - This investigation assessed the frequency of catamenial epilepsy in 87 women who charted seizures and menses during three cycles. Catamenial epilepsy designation was made if two of three cycles showed at least one of three previously defined catamenial patterns. Among ovulatory cycles, average daily seizure frequency was significantly greater during the perimenstrual and preovulatory phases. Among anovulatory cycles, average daily seizure frequency was substantially less during the midfollicular phase than during the remainder of the cycle. Overall, 39.1% of the women had catamenial epilepsy. PMID- 15349873 TI - Hemispheric mediation of spatial attention: pseudoneglect after callosal stroke. AB - Study of patients with callosal lesions can provide insight into the mediation of spatial attention-intention by each hemisphere. Two patients with anterior callosal strokes bisected lines to the left of midline with the left hand and to the right of midline with the right in both a visual and tactile bisection task. The patients demonstrated a similar pattern of performance on pointing to body midline in space. These results are consistent with the notion that each hemisphere supports spatial attention-intention toward contralateral space and that the corpus callosum is critical in the integration of such information. PMID- 15349874 TI - Glucose hypometabolism of hypothalamus and thalamus in narcolepsy. AB - It has been hypothesized that hypothalamus is involved in narcolepsy. The relative difference between cerebral glucose metabolism of 24 narcoleptic patients and 24 normal controls was studied using 18F-fluorodeoxy glucose positron emission tomography. Patients with narcolepsy showed significantly reduced cerebral glucose metabolism in bilateral rectal and subcallosal gyri, the medial convexity of right superior frontal gyrus, bilateral precuneus, right inferior parietal lobule, and in left supramarginal gyrus (uncorrected p < 0.001). Bilateral posterior hypothalami and mediodorsal thalamic nuclei showed hypometabolism with significance at the level of corrected p < 0.05, with small volume correction. This study showed cerebral glucose hypometabolism of the hypothalamus-thalamus-orbitofrontal pathways in the narcoleptic brain. PMID- 15349875 TI - Mechanism of action of voltage-gated K+ channel antibodies in acquired neuromyotonia. AB - Acquired neuromyotonia (ANM) is associated with antibodies to voltage-gated K+ channels (VGKCs). ANM sera reduce the number of K+ currents in neuronal cell lines, but it is not clear how the antibodies act. Here, we show by using the NB 1 cell line that the reduction in K+ currents by IgG is independent of added complement. IgG Fc and Fab fragments from ANM sera had no effect, but three of four ANM F(ab')2 fragments significantly reduced K+ currents. Thus, cross-linking of the channels by divalent antibodies is likely to be an important mechanism in reducing K+ currents. PMID- 15349876 TI - Saccade velocity is controlled by polyglutamine size in spinocerebellar ataxia 2. AB - We assessed maximal saccade velocity (MSV) in 82 spinocerebellar ataxia type 2 (SCA2) patients and 80 controls, correlating it to disease duration, polyglutamine expansion size, age at onset, ataxia score, age, and sex. Little overlap with normal values was found even at earliest stages. Stepwise linear regression analysis showed that 60-degree MSV was strongly influenced by polyglutamine size and less by disease duration, whereas the reverse was found for ataxia score. Saccade velocity thus is a sensitive, quite specific, and objective endophenotype, useful to search polyglutamine modifier genes. PMID- 15349877 TI - Ataxin-7 aggregation and ubiquitination in infantile SCA7 with 180 CAG repeats. AB - Extremely long (>150) CAG repeats are often used to create models of polyglutamine diseases yet are very rare in humans where they manifest as pediatric multisystem syndromes of little specificity. Here, we describe an infant with 180 CAG repeats in the spinocerebellar ataxia type 7 gene and focus on systemic ataxin-7 aggregation. This was found in many organs, including the cardiovascular system. In the brain, the hippocampus emerged as a principal site of ataxin-7 aggregation without cell loss. We note differential ubiquitination of aggregates and discuss how this may relate to selective vulnerability. PMID- 15349878 TI - White matter volume increase and minicolumns in autism. PMID- 15349879 TI - Sequence analysis of familial PEO shows additional mutations associated with the 752C-->T and 3527C-->T changes in the POLG1 gene. PMID- 15349881 TI - A de novo LGI1 mutation in sporadic partial epilepsy with auditory features. PMID- 15349883 TI - Advertising in Hepatology. PMID- 15349887 TI - Funding for research in hepatology. PMID- 15349886 TI - Hepatitis C infection and injection drug use: the role of hepatologists in evolving treatment efforts. AB - Treatment regimens for both substance abuse and hepatitis C infection are complex and evolving. New pharmacotherapy for opioid addiction allows for office-based treatment and, thus, an opportunity for expanded treatment in the context of hepatitis C infection. The current article addresses the newly evolving, complex issues in the medical management of hepatitis C and injection drug use. PMID- 15349888 TI - A new way to look at liver cancer. PMID- 15349889 TI - Is severe recurrent hepatitis C more common after adult living donor liver transplantation? PMID- 15349890 TI - TNF-alpha regulates mouse fetal hepatic maturation induced by oncostatin M and extracellular matrices. AB - Fetal hepatic maturation consists of multisteps and is regulated by several cytokines and cell-cell or cell-matrices interactions. In the mid-to-late fetal stage, hepatocytes have few metabolic functions associated with adult liver homeostasis. Cultured fetal hepatocytes acquire the expression of several mature liver-specific genes through stimulation with hepatic maturation factor oncostatin M (OSM) and matrigel. Tumor necrosis factor-alpha (TNFalpha) regulates fetal hepatic maturation stimulated by OSM and matrigel. TNFalpha suppressed expression of mature liver-specific genes such as tyrosine aminotransferase and apolipoproteins. In addition, the expression of hematopoietic cytokines and cyclin A2, repressed by OSM and matrigel, is induced by TNFalpha in the fetal hepatic cultures coincident with cell division. TNFalpha inhibited the induction of hepatocyte nuclear factor 4alpha induced by OSM and matrigel, suggesting that down-regulation of hepatocyte nuclear factor 4alpha expression is involved in the mechanism of suppression of hepatic maturation by TNFalpha. Interestingly, TNFalpha is expressed in the prenatal and postnatal liver but not in adult liver, whereas TNFR1, a TNFalpha receptor, is expressed in both fetal and adult livers. In conclusion, TNFalpha is a suppressive factor of hepatic maturation. The balance between hepatic maturation factor (OSM and extracellular matrices) and TNFalpha is important for liver development. PMID- 15349891 TI - Rapamycin-sensitive induction of eukaryotic initiation factor 4F in regenerating mouse liver. AB - Following acute injuries that diminish functional liver mass, the remaining hepatocytes substantially increase overall protein synthesis to meet increased metabolic demands and to allow for compensatory liver growth. Previous studies have not clearly defined the mechanisms that promote protein synthesis in the regenerating liver. In the current study, we examined the regulation of key proteins involved in translation initiation following 70% partial hepatectomy (PH) in mice. PH promoted the assembly of eukaryotic initiation factor (eIF) 4F complexes consisting of eIF4E, eIF4G, eIF4A1, and poly-A binding protein. eIF4F complex formation after PH occurred without detectable changes in eIF4E-binding protein 1 (4E-BP1) phosphorylation or its binding eIF4E. The amount of serine 1108-phosphorylated eIF4G (but not Ser209-phosphorylated eIF4E) was induced following PH. These effects were antagonized by treatment with rapamycin, indicating that target of rapamycin (TOR) activity is required for eIF4F assembly in the regenerating liver. Rapamycin inhibited the induction of cyclin D1, a known eIF4F-sensitive gene, at the level of protein expression but not messenger RNA (mRNA) expression. In conclusion, increased translation initiation mediated by the mRNA cap-binding complex eIF4F contributes to the induction of protein synthesis during compensatory liver growth. Further study of factors that regulate translation initiation may provide insight into mechanisms that govern metabolic homeostasis and regeneration in response to liver injury. PMID- 15349892 TI - Exploring interactions between rat hepatocytes and nonparenchymal cells using gene expression profiling. AB - Cocultivation of primary hepatocytes with a plethora of nonparenchymal cells (from within and outside the liver) has been shown to support hepatic functions in vitro. Despite significant investigation into this phenomenon, the molecular mechanism underlying epithelial-nonparenchymal interactions in hepatocyte cocultures remains poorly understood. In this study, we present a functional genomic approach utilizing gene expression profiling to isolate molecular mediators potentially involved in induction of liver-specific functions by nonparenchymal cells. Specifically, primary rat hepatocytes were cocultivated with closely related murine fibroblast cell types (3T3-J2, NIH-3T3, mouse embryonic fibroblasts) to allow their classification as "high," "medium," or "low" inducers of hepatic functions. These functional responses were correlated with fibroblast gene expression profiles obtained using Affymetrix GeneChips. Microarray data analysis provided us with 17 functionally characterized candidate genes in the cell communication category (cell surface, extracellular matrix, secreted factors) that may be involved in induction of hepatic functions. Further analysis using various databases (i.e., PubMed, GenBank) facilitated prioritization of the candidates for functional characterization. We experimentally validated the potential role of two candidates in our coculture model. The cell surface protein, neural cadherin (N-cadherin), was localized to hepatocyte-fibroblast junctions, while adsorbed decorin up-regulated hepatic functions in pure cultures as well as cocultures with low-inducing fibroblasts. In the future, identifying mediators of hepatocyte differentiation may have implications for both fundamental hepatology and cell-based therapies (e.g., bioartificial liver devices). In conclusion, the functional genomic approach presented in this study may be utilized to investigate mechanisms of cell-cell interaction in a variety of tissues and disease states. PMID- 15349893 TI - Selective priming to Toll-like receptor 4 (TLR4), not TLR2, ligands by P. acnes involves up-regulation of MD-2 in mice. AB - Lipopolysaccharide (LPS) triggers cytokine production through Toll-like receptor 4 (TLR4), which shares downstream signaling pathways with TLR2. We investigated the roles of TLR2 and TLR4 in Propionibacterium acnes (P. acnes)-primed, LPS induced liver damage using selective TLR ligands. Stock LPS induced interleukin 8 in both TLR4- and TLR2-expressing human embryonic kidney (HEK) 293 cells. Purified LPS (TLR4 ligand) activated HEK/TLR4 cells, while peptidoglycan and lipoteichoic acid (TLR2 ligands) activated HEK/TLR2 cells, respectively. In mice, P. acnes priming resulted in increased liver messenger RNA (mRNA) and serum levels of tumor necrosis factor alpha, interleukin 12, and interferon gamma (IFN gamma) by both stock LPS and purified LPS challenges compared with nonprimed controls. In contrast, P. acnes failed to sensitize to TLR2 ligands (peptidoglycan + lipoteichoic acid). In the liver, P. acnes-priming was associated with up-regulation of TLR4 and MD-2 proteins, and subsequent LPS challenge further increased MD-2 and CD14 mRNA levels. The lack of sensitization to TLR2 ligands by P. acnes correlated with no increase in hepatic TLR1 or TLR6 mRNA. In vitro, P. acnes pretreatment desensitized RAW macrophages to a secondary stimulation via both TLR2 and TLR4. However, IFN-gamma could selectively prevent desensitization to TLR4 but not to TLR2 ligands. Furthermore, P. acnes induced production of IFN-gamma in vivo as well as in isolated splenocytes. In vitro, P. acnes-primed Hepa 1-6 hepatocytes but not RAW macrophages produced increased MD-2 and CD14 mRNA levels after an LPS challenge. In conclusion, P. acnes priming to selective TLR4-mediated liver injury is associated with up-regulation of TLR4 and MD-2 and is likely to involve IFN-gamma and prevent TLR4 desensitization by P. acnes. PMID- 15349894 TI - The role of iNOS in alcohol-dependent hepatotoxicity and mitochondrial dysfunction in mice. AB - Nitric oxide (NO) is now known to control both mitochondrial respiration and organelle biogenesis. Under conditions of ethanol-dependent hepatic dysfunction, steatosis is increased, and this is associated with increased expression of inducible nitric oxide synthase (iNOS). We have previously shown that after chronic exposure to ethanol, the sensitivity of mitochondrial respiration to inhibition by NO is enhanced, and we have proposed that this contributes to ethanol-dependent hypoxia. This study examines the role of iNOS in controlling the NO-dependent modification of mitochondrial function. Mitochondria were isolated from the livers of both wild-type (WT) and iNOS knockout (iNOS-/-) mice that were fed an isocaloric ethanol-containing diet for a period of 5 weeks. All animals that consumed ethanol showed some evidence of fatty liver; however, this was to a lesser extent in the iNOS-/- mice compared to controls. At this early stage in ethanol-dependent hepatic dysfunction, infiltration of inflammatory cells and the formation of nitrated proteins was also decreased in response to ethanol feeding in the iNOS-/- animals. Mitochondria isolated from wild-type ethanol-fed mice showed a significant decrease in respiratory control ratio and an increased sensitivity to NO-dependent inhibition of respiration relative to their pair-fed controls. In contrast, liver mitochondria isolated from iNOS-/- mice fed ethanol showed no change in the sensitivity to NO-dependent inhibition of respiration. In conclusion, the hepatic response to chronic alcohol-dependent cytotoxicity involves a change in mitochondrial function dependent on the induction of iNOS. PMID- 15349895 TI - Peroxynitrite alters the catalytic activity of rodent liver proteasome in vitro and in vivo. AB - The proteasome is an important multicatalytic enzyme complex that degrades misfolded and oxidized proteins, signal transduction factors, and antigenic peptides for presentation. We investigated the in vitro effects of peroxynitrite (PN) on the peptidase activity of both crude 20S and 26S and purified 20S proteasome preparations from rat liver as well as proteasome activity in Hep G2 cells and in mouse liver. Crude and purified proteasome preparations were exposed to PN or to the PN donor, 3-morpholinosydnonimine hydrochloride (SIN-1), and then assayed for chymotrypsin-like activity. For in vivo experiments, mice were treated with molsidomine, which is metabolized to SIN-1 in liver. PN and SIN-1 dose-dependently modulated the chymotrypsin-like activity of the 20S proteasome: lower concentrations enhanced proteasome activity, and higher concentrations caused its decline. The NO donor S-nitroso-N-acetylpenicillamine (SNAP), at all concentrations, suppressed 20S proteasome activity. We observed similar results when liver soluble fractions (S-100) were treated with PN, SIN-1, or SNAP, except that enzyme activity in S-100 fractions was less sensitive than the purified enzymes to these agents. Treatment of Hep G2 cells with 0.01 or 0.1 mmol/L SIN-1 stimulated in situ proteasome activity in these cells, while 1 mmol/L SIN-1 suppressed it. SNAP treatment did not affect proteasome activity in Hep G2 cells. Mice treated with molsidomine had enhanced liver proteasome activity 6 hours after treatment, but after 24 hours enzyme activity declined below control levels. In conclusion, PN dose-dependently modulated proteasome activity, regulating protein degradation by the proteasome in liver cells. PMID- 15349896 TI - Role of the renin-angiotensin system in hepatic ischemia reperfusion injury in rats. AB - It has been shown that the renin-angiotensin system (RAS) plays key roles in the development of fibrosis in numerous organs, including the liver. Other studies have suggested that the RAS also may play roles in diseases of chronic inflammation. However, whether the RAS also can mediate acute inflammation in liver is unclear. The purpose of this study therefore was to determine the effect of the RAS inhibitors captopril and losartan on acute liver damage and inflammation caused by hepatic ischemia and subsequent reperfusion. Accordingly, male rats were subjected to 1 hour of hepatic ischemia (70%) followed by reperfusion; animals were killed 3, 8, or 24 hours after reperfusion. The effect of captopril or losartan (100 or 5 mg/kg intragastrically, respectively) was compared with that of vehicle (saline). The expression of angiotensinogen in liver increased fivefold 3 hours after reperfusion. Indices of liver damage and inflammation (e.g., alanine aminotransferase levels, pathological features, tumor necrosis factor-alpha levels, and intercellular adhesion molecule-1 expression) all were significantly elevated in vehicle-treated animals after hepatic ischemia and subsequent reperfusion. Ischemia and reperfusion also caused an increase in the accumulation of protein adducts of 4-hydroxynonenal, an index of oxidative stress. Captopril or losartan treatment showed profound protective effects under these conditions, significantly blunting the increase in all these parameters caused by ischemia and reperfusion. In conclusion, RAS inhibitors prevent acute liver injury in a model of inflammation caused by ischemia and reperfusion. These data further suggest that the RAS may play a key role in mediating such responses in the liver and suggest a novel role for this system. PMID- 15349897 TI - In vitro differentiation of rat liver derived stem cells results in sensitization to TNFalpha-mediated apoptosis. AB - Hepatic stem cells are activated after liver damage and have a critical role in tissue homeostasis and repair. Characterization of molecular and cellular events accompanying the expansion and differentiation of liver stem cells is essential for understanding the basic biology of stem cells and for facilitating clinical application of the stem cells. We assessed whether in vitro differentiation of putative hepatic progenitor (rat liver epithelial [RLE]) cells toward hepatocytic lineage affects the response to TNFalpha-mediated cytotoxicity, a common determinant of liver injury. The data show that 50% of differentiated cells underwent apoptosis after 6 hours of TNFalpha treatment whereas control RLE cells were resistant. Both cell types displayed mitochondrial depolarization and release of cytochrome c but the TNFalpha treatment resulted in activation of caspases 9 and 3 and the execution of apoptosis only in differentiated RLE cells. Apoptotic death was associated with increased ROS production and depletion of glutathione. Antioxidants completely prevented both glutathione depletion and apoptosis induced by TNFalpha in differentiated RLE cells. Conversely, glutathione-depleting agents sensitized control RLE cells to TNFalpha induced apoptosis. In conclusion, efficient antioxidant defense system involving glutathione renders hepatic progenitor cells resistant to TNFalpha-mediated apoptosis and acquisition of sensitivity to death stimuli is an implicit feature of the differentiation process. Supplementary material for this article can be found on the HEPATOLOGY website (http://interscience.wiley.com/jpages/0270 9139/suppmat/index.html). PMID- 15349898 TI - In vivo regulation of murine CYP7A1 by HNF-6: a novel mechanism for diminished CYP7A1 expression in biliary obstruction. AB - Disruption of the enterohepatic bile acid circulation during biliary tract obstruction leads to profound perturbation of the cholesterol and bile acid metabolic pathways. Several families of nuclear receptor proteins have been shown to modulate this critical process by regulating hepatic cholesterol catabolism and bile acid synthesis through the transcriptional control of cholesterol 7 alpha hydroxylase (CYP7A1). Hepatocyte nuclear factor (HNF) 6 (also known as OC 1) is a member of the ONECUT family of transcription factors that activate numerous hepatic target genes essential to liver function. We have previously shown that hepatic expression of mouse HNF-6 messenger RNA (mRNA) and protein significantly decrease following bile duct ligation. Because CYP7A1 contains potential HNF-6 binding sites in its promoter region, we tested the hypothesis that HNF-6 transcriptionally regulates CYP7A1. Following bile duct ligation, we demonstrated that diminished HNF-6 mRNA levels correlate with a reduction in CYP7A1 mRNA expression. Increasing hepatic levels of HNF-6 either by infection with recombinant adenovirus vector expressing HNF-6 cDNA by growth hormone treatment leads to an induction of CYP7A1 mRNA. To directly evaluate if HNF-6 is a transcriptional activator for CYP7A1, we used deletional and mutational analyses of CYP7A1 promoter sequences and defined sequences -206/-194 to be critical for CYP7A1 transcriptional stimulation by HNF-6 in cotransfection assays. In conclusion, the HNF-6 protein is a component of the complex network of hepatic transcription factors that regulates the expression of hepatic genes essential for bile acid homeostasis and cholesterol/lipid metabolism in normal and pathological conditions. PMID- 15349899 TI - Proteomic signature corresponding to alpha fetoprotein expression in liver cancer cells. AB - Alpha fetoprotein (AFP) has been implicated in the development of hepatocellular carcinoma and is considered to be a diagnostic and prognostic tumor marker. Because elevated expression of AFP is associated with many characteristics of hepatocellular carcinoma tissues, we hypothesized that multiple proteins may function in a coordinated manner with AFP. To identify such proteins, we performed global protein expression analysis, namely a proteomic study. The protein expression profiles of 9 AFP-producing liver cancer cell lines (JHH-5, HuH-1, PLC/PRL/5, Hep3B, HT-17, JHH-7, HuH-7, HepG2, Li-7) and 7 nonproducing liver cancer cell lines (HLE, JHH-6, Sk-Hep-1, JHH-4, HLF, RBE, SSP-25) were generated by fluorescence 2-dimensional difference gel electrophoresis. In fluorescence 2-dimensional difference gel electrophoresis, proteins are labeled with fluorescent dyes before electrophoresis for more accurate quantitative expression analysis. We identified 11 protein spots that distinguished AFP producing cell lines from nonproducing cell lines by multivariate studies. The spots showed consistent alterations in amount in AFP-producing cell lines (6 up regulated and 5 down-regulated). An additional 5 liver cancer cell lines (KIM-1, KYN-2, KYN-3, PH5-CH, PH5-T) also were correctly grouped with respect to their AFP production on the basis of the intensity of the 11 protein spots. The proteins corresponding to the 11 selected spots were identified by mass spectrometry and were categorized into 4 groups based on their known role in apoptosis, glucose metabolism, cytoskeletal organization, or translation. In conclusion, we found a novel association of AFP with other proteins. Their interaction should provide insight into the biology of AFP-producing hepatocellular carcinoma cells. PMID- 15349900 TI - Negative regulation of hepatocellular carcinoma cell growth by signal regulatory protein alpha1. AB - Signal regulatory protein (SIRP) alpha1 is a member of the SIRP family that undergoes tyrosine phosphorylation and binds SHP-2 tyrosine phosphatase in response to various mitogens. The expression levels of SIRPalpha1 were decreased in HCC tissues, compared with the matched normal tissues. Exogenous expression of wild type SIRPalpha1, but not of a mutant SIRPalpha1 lacking the tyrosine phosphorylation sites, in SIRPalpha1-negative Huh7 human HCC cells resulted in suppression of tumor cell growth both in vitro and in vivo. Treatment of Huh7 transfectants with EGF or HGF induced tyrosine phosphorylation of SIRPalpha1 and its association with SHP-2, which were accompanied by reduced ERK1 activation. Expression of SIRPalpha1 significantly suppressed activation of NF-kappaB and also sensitized Huh7 cells to TNFalpha or cisplatin-induced cell death. In addition, SIRPalpha1-transfected Huh7 cells displayed reduced cell migration and cell spreading in a fashion that was dependent on SIRPalpha1/SHP-2 complex formation. In conclusion, a negative regulatory effect of SIRPalpha1 on hepatocarcinogenesis is exerted, at least in part, through inhibition of ERK and NF-kappaB pathways. PMID- 15349901 TI - Randomized controlled study of TIPS versus paracentesis plus albumin in cirrhosis with severe ascites. AB - The transjugular intrahepatic portosystemic shunt (TIPS) has been shown to be effective in the control of refractory or recidivant ascites. However, the effect of TIPS on survival as compared with that of large-volume paracentesis plus albumin is uncertain. A multicenter, prospective, clinical trial was performed in 66 patients with cirrhosis and refractory or recidivant ascites (16 Child Turcotte-Pugh class B and 50 Child-Turcotte-Pugh class C) randomly assigned to treatment with TIPS (n = 33) or with large-volume paracentesis plus human albumin (n = 33). The primary endpoint was survival without liver transplantation. Secondary endpoints were treatment failure, rehospitalization, and occurrence of complications. Thirteen patients treated with TIPS and 20 patients treated with paracentesis died during the study period, 4 patients in each group underwent liver transplantation. The probability of survival without transplantation was 77% at 1 year and 59% at 2 years in the TIPS group as compared with 52% and 29% in the paracentesis group (P = .021). In a multivariate analysis, treatment with paracentesis and higher MELD score showed to independently predict death. Treatment failure was more frequent in patients assigned to paracentesis, whereas severe episodes of hepatic encephalopathy occurred more frequently in patients assigned to TIPS. The number and duration of rehospitalizations were similar in the two groups. In conclusion, compared to large-volume paracentesis plus albumin, TIPS improves survival without liver transplantation in patients with refractory or recidivant ascites. PMID- 15349902 TI - Serum from patients with fulminant hepatic failure causes hepatocyte detachment and apoptosis by a beta(1)-integrin pathway. AB - Hepatocyte transplantation is restricted by the impaired ability of hepatocytes to engraft and survive in the damaged liver. Understanding the mechanisms that control this process will permit the development of strategies to improve engraftment. We studied changes in liver matrix during acute injury and delineated the mechanisms that perturb the successful adhesion and engraftment of hepatocytes. Collagen IV expression was increased in sinusoidal endothelium and portal tracts of fulminant hepatic failure explants, whereas there were minimal changes in the expression of fibronectin, tenascin, and laminin. Using an in vitro model of cellular adhesion, hepatocytes were cultured on collagen-coated plates and exposed to serum from patients with liver injury to ascertain their subsequent adhesion and survival. There was a rapid, temporally progressive decrease in the adhesive properties of hepatocytes exposed to such serum that occurred within 4 hours of exposure. Loss of activity of the beta1-integrin receptor, which controls adhesion to collagen, was seen to precede this loss of adhesive ability. Addition of the beta1-integrin activating antibody (TS2/16) to cells cultured with liver injury serum significantly increased their adhesion to collagen, and prevented significant apoptosis. In conclusion, we have identified an important mechanism that underpins the failure of infused hepatocytes to engraft and survive in liver injury. Pretreating cells with an activating antibody can improve their engraftment and survival, indicating that serum from patients with liver injury exerts a defined nontoxic biological effect. This finding has important implications in the future of cellular transplantation for liver and other organ diseases. PMID- 15349903 TI - Effects of contrast media on renal function in patients with cirrhosis: a prospective study. AB - Patients with cirrhosis are frequently submitted to radiological procedures that require the administration of contrast media. Contrast media is a well-known cause of renal failure, particularly in the presence of some predisposing conditions. However, it is not known whether cirrhosis constitutes a risk factor for contrast media-induced renal failure. The aim of this study was to assess the possible nephrotoxicity of contrast media in patients with cirrhosis. In a first protocol, renal function was evaluated with sensitive methods (glomerular filtration rate using iothalamate I 125 clearance and renal plasma flow using iodohippurate I 131 clearance) before and 48 hours after the administration of contrast media in 31 patients with cirrhosis (20 with ascites, 5 with renal failure). Solute-free water clearance, urine sodium, prostaglandins, and markers of tubular damage were also measured. The administration of contrast media was not associated with significant changes in renal function tests, neither in the whole group of patients nor in patients with ascites or renal failure. Urinary prostaglandin E2 and N-acetyl-beta-D-glucosaminidase increased significantly, but sodium and solute-free water excretion remained unchanged. In a second protocol, a different series of 60 patients with cirrhosis and renal failure were examined prospectively. No patient had renal failure due to contrast media. Only in 1 patient with septic shock was contrast media a possible contributing factor. In conclusion, the administration of contrast media is not associated with adverse effects on renal function in patients with cirrhosis. Cirrhosis does not appear to be a risk factor for the development of contrast media-induced nephrotoxicity. PMID- 15349904 TI - Improved survival after variceal bleeding in patients with cirrhosis over the past two decades. AB - Over the past two decades, new treatment modalities have been introduced for the management of variceal bleeding. The aim of this retrospective study in a single center was to assess whether these treatments have improved the prognosis for cirrhotic patients with variceal bleeding. We reviewed the clinical records of all patients with cirrhosis admitted to our Liver Intensive Care Unit due to variceal bleeding during the years 1980, 1985, 1990, 1995, and 2000. Whereas balloon tamponade was still the first-line treatment in 1980, patients treated in 2000 received a vasoactive agent, an endoscopic treatment, and an antibiotic prophylaxis in, respectively, 90%, 100%, and 94% of cases. The in-hospital mortality rate steadily decreased over the study period: 42.6%, 29.9%, 25%, 16.2%, and 14.5% in 1980, 1985, 1990, 1995, and 2000, respectively (P < .05). Mortality decreased from 9% in 1980 to 0% in 2000 in Child-Turcotte-Pugh class A patients, from 46% to 0% in class B patients, and from 70% to 32% in class C patients. This improved survival was associated with a decrease of rebleeding (from 47% in 1980 to 13% in 2000) and bacterial infection rates (from 38% to 14%). On multivariable analysis, endoscopic therapy and antibiotic prophylaxis were independent predictors of survival. In conclusion, in-hospital mortality of patients with cirrhosis and variceal bleeding decreased threefold over the past two decades, in concurrence with an early and combined use of pharmacological and endoscopic therapies and short-term antibiotic prophylaxis. PMID- 15349905 TI - Gas exchange mechanism of orthodeoxia in hepatopulmonary syndrome. AB - The mechanism of orthodeoxia (OD), or decreased partial pressure of arterial oxygen (PaO2) from supine to upright, a characteristic feature of hepatopulmonary syndrome (HPS), has never been comprehensively elucidated. We therefore investigated the intrapulmonary (shunt and ventilation-perfusion [VA/Q] mismatching) and extrapulmonary factors governing PaO2 in 20 patients with mild to severe HPS (14 males, 6 females; 50 +/- 3 years old SE) at upright and supine, in random order. We set out a cutoff value for OD, namely a PaO2 decrease > or = 5% or > or = 4 mm Hg (area under the receiver operating characteristic curve, 0.96 each). Compared to supine, 5 patients showed OD (PaO2 change, -11% +/- 2%, 7 +/- 1 mm Hg, P < .05) with further VA/Q worsening (shunt + low VA/Q mode increased from 19% +/- 7% to 21% +/- 7% of cardiac output [QT], P < .05), as opposed to 15 patients who did not (+2% +/- 2%, +1+/- 1 mm Hg) with VA/Q improvement (from 20% +/- 4% to 16% +/- 4% of QT, P < .01). Cardiac output was significantly lower in OD patients in both positions. Changes in extrapulmonary factors at upright, such as increased minute ventilation and decreased QT, were of similar magnitude in both subsets of patients. In conclusion, our data suggest that gas exchange response to OD in HPS points to a more altered pulmonary vascular tone inducing heterogeneous blood flow redistribution to lung zones with prominent intrapulmonary vascular dilatations. PMID- 15349906 TI - Classification and prediction of survival in hepatocellular carcinoma by gene expression profiling. AB - We analyzed global gene expression patterns of 91 human hepatocellular carcinomas (HCCs) to define the molecular characteristics of the tumors and to test the prognostic value of the expression profiles. Unsupervised classification methods revealed two distinctive subclasses of HCC that are highly associated with patient survival. This association was validated via 5 independent supervised learning methods. We also identified the genes most strongly associated with survival by using the Cox proportional hazards survival analysis. This approach identified a limited number of genes that accurately predicted the length of survival and provides new molecular insight into the pathogenesis of HCC. Tumors from the low survival subclass have strong cell proliferation and antiapoptosis gene expression signatures. In addition, the low survival subclass displayed higher expression of genes involved in ubiquitination and histone modification, suggesting an etiological involvement of these processes in accelerating the progression of HCC. In conclusion, the biological differences identified in the HCC subclasses should provide an attractive source for the development of therapeutic targets (e.g., HIF1a) for selective treatment of HCC patients. Supplementary material for this article can be found on the HEPATOLOGY Web site (http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html) PMID- 15349907 TI - Reduced expression of cell cycle regulator p18(INK4C) in human hepatocellular carcinoma. AB - Cyclins, cyclin-dependent kinases (Cdks), and Cdk inhibitors (CdkIs) are frequently altered in human cancer. p18INK4C, a member of the INK4 family of CdkIs, is a potential tumor-suppressor gene product. However, the expression of p18INK4C in hepatocellular carcinoma (HCC) remains unknown. The aim of this study was to examine the expression of p18INK4C in various liver diseases including HCC and to assess its clinical significance in HCC. To that end, we examined the expression of p18INK4C by immunohistochemistry in various liver diseases, including 51 HCCs, and also studied the relationship between p18INK4C expression, the phosphorylation of retinoblastoma protein (pRb), and the activity level of Cdk4 and Cdk6. Immunohistochemical analysis revealed the frequent loss of p18INK4C expression in HCC, especially in poorly differentiated HCC. The loss of p18INK4C expression was shown to be associated with a poor prognosis compared with that associated with p18INK4C- positivity. Further, the kinase activity of Cdk4 was found to be higher in p18INK4C-negative HCCs than in p18INK4C- positive HCCs. However, the level of Cdk6 activity was similar in the 2 groups of HCCs. In p18INK4C- positive HCCs, p18INK4C dominantly interacted with Cdk4 rather than with Cdk6. pRb phosphorylated at serine(Ser) 780 was detected more frequently in p18INK4C - negative than in p18INK4C - positive HCCs. In conclusion, the loss of p18INK4C expression may play a role in the differentiation and development of HCC through the up-regulation of Cdk4 activity. PMID- 15349908 TI - A novel IgM class autoantibody to a hepatocyte-related 190 kDa molecule in patients with type 1 autoimmune hepatitis. AB - It has been reported that autoantibodies to hepatocytes are frequently found in patients with autoimmune hepatitis (AIH). To elucidate the nature of these hepatocyte-specific autoantibodies, we attempted to generate a hepatocyte specific monoclonal antibody (MoAb) from Epstein-Barr virus-transformed peripheral blood mononuclear cells obtained from a patient with AIH. We established a single clone, 2E3, that continued to produce an immunoglobulin M (IgM) antibody (lambda-type). This MoAb had the following properties: it reacted mainly with hepatocyte-derived cell lines, rather than with other cell lines, and it reacted with liver tissue but not with other tissues. By immunoblot analysis, we found that this MoAb recognized a 190 kDa molecule on hepatocytes. The MoAb was able to kill hepatocyte-derived cell lines in the presence of fresh human serum. This cytotoxic effect was completely abrogated by heat inactivation of human serum prior to its addition to cell lines. In addition, an IgM autoantibody that recognized a 190 kDa molecule was also found in patients with AIH but not in those with chronic hepatitis C; its titer correlated significantly with serum alanine aminotransferase (ALT) levels in patients with AIH. In conclusion, we generated a human MoAb that recognizes a 190 kDa molecule on hepatocytes. Because of its ability to mediate complement-dependent cytotoxicity and the presence of similar IgM autoantibody in patients with AIH, we hypothesize this autoantibody may play a role in the immunopathogenesis of AIH. PMID- 15349909 TI - Effect of high-dose ursodeoxycholic acid on its biliary enrichment in primary sclerosing cholangitis. AB - Ursodeoxycholic acid (UDCA) has beneficial effects in cholestatic liver diseases. In primary sclerosing cholangitis (PSC), there is evidence that high doses (+/- 20 mg/kg) of UDCA may be more effective than average doses. Biliary enrichment of UDCA at such high doses may represent the decisive factor for its beneficial effect. Up to now it is not clear how high-dose UDCA correlates with its biliary enrichment and whether bacterial degradation of large amounts of UDCA may lead to an increased bacterial formation of more toxic hydrophobic bile acids. We determined the biliary bile acid composition in 56 patients with PSC including 30 patients with repeat bile samples treated with various doses of UDCA. At a UDCA dose of 10-13 mg/kg/d (n = 18) biliary UDCA represented 43.1% + 0.3% (mean + SD) of total bile acids; at a UDCA dose of 14-17 mg/kg (n = 14), its biliary content increased to 46.9% + 0.3%, at 18-21 mg/kg (n = 34) to 55.9% + 0.2%, at 22-25 mg/kg (n = 12) to 58.6% + 2.3%, and at 26-32 mg/kg (n = 8) to 57.7% + 0.4%. During UDCA treatment, the biliary content of all other bile acids was unchanged or decreased. In conclusion, biliary enrichment of UDCA increases with increasing dose and reaches a plateau at 22-25 mg/kg. There was no increase of toxic hydrophobic bile acids. If biliary enrichment of UDCA represents the decisive factor for its clinical effect, it seems likely that UDCA doses of up to 22-25 mg/kg may be more effective than lower doses. PMID- 15349910 TI - Hepatitis C recurrence is more severe after living donor compared to cadaveric liver transplantation. AB - Preliminary reports suggested that hepatitis C virus (HCV) infection has a more aggressive course following living donor liver transplantation (LDLT) compared to cadaveric liver transplantation (CLT). The aim of this prospective study was to establish if HCV disease recurrence differs between LDLT and CLT. A cohort of 116 consecutive HCV-infected patients undergoing 117 LTs in a single center from March 2000 to August 2003 were followed-up, including systematic liver biopsies. Severe recurrence (SR) was defined as biopsy-proven cirrhosis and/or the occurrence of clinical decompensation. After a median follow-up of 22 months (2.6 44 months), 26 (22%) patients developed SR (decompensation in 12), involving 17 (18%) of 95 patients undergoing CLT and 9 (41%) of 22 undergoing LDLT. The 2-year probability of presenting SR was significantly higher in LDLT compared to CLT (45% vs. 22%, P = .019). By univariate analysis LDLT (P = .019) and an ALT higher than 80 IU/L 3 months after LT (P = .022) were predictors of SR. In 93 patients from whom a liver biopsy was available 3 months after LT, a lobular necroinflammatory score >1 (P < .01), LDLT (P < .01), and biliary complications (P = .046) were associated with SR. However, the only variables independently associated with SR were LDLT (odds ratio [OR], = 2.8; 95% CI,1.19-6.6; P = .024) and a lobular necroinflammatory score > 1 (OR, 3.1; 95% CI, 1.2-8; P = .013). In conclusion, HCV recurrence is more severe in LDLT compared to CLT. Although our results were based on a single-center experience, they should be considered in the decision-making process of transplant programs, since severe HCV recurrence may ultimately compromise graft and patient survival. PMID- 15349911 TI - Hepatitis C virus NS5A-regulated gene expression and signaling revealed via microarray and comparative promoter analyses. AB - Most individuals exposed to hepatitis C virus (HCV) become chronically infected and are predisposed to liver disease. The mechanisms underlying viral persistence and disease progression are unknown. A role for the HCV NS5A protein in viral replication and interferon resistance has been demonstrated. To identify mechanisms affected by NS5A, we analyzed the gene expression of Huh7 cells expressing NS5A and control cells using oligonucleotide microarrays. A set of 103 genes (43 up-regulated, 60 down-regulated) whose expression was modified by at least twofold was selected. These included genes involved in cell adhesion and motility, calcium homeostasis, lipid transport and metabolism, and genes regulating immune responses. The finding of modulated expression of genes related to the TGF-beta superfamily and liver fibrosis was observed. Interestingly, both the tumor necrosis factor and lymphotoxin beta receptors were down-regulated by NS5A. Similar data were obtained following expression of four NS5A mutants obtained from patients who were not responsive or were sensitive to interferon therapy. Through computational analysis, we determined that 39 of the 43 genes up regulated by NS5A contained one or more nuclear factor kappaB (NF-kappaB) binding sites within their promoter region. Using the Gibbs sampling method, we also detected enrichment of NF-kappaB consensus binding sites in the upstream regions of the 43 coexpressed genes. Activation of NF-kappaB by NS5A was subsequently demonstrated in luciferase reporter assays. Adenovirus-mediated expression of IkappaBalpha reverted NS5A mediated up-regulation of gene expression. In conclusion, this study suggests a role of NS5A and NF-kappaB in HCV pathogenesis and related liver disease. Supplementary material for this article can be found on the HEPATOLOGY website (http://interscience.wiley.com/jpages/0270 9139/suppmat/index.html). PMID- 15349912 TI - A dose-finding study of once-daily oral telbivudine in HBeAg-positive patients with chronic hepatitis B virus infection. AB - Current therapy for chronic hepatitis B is suboptimal as a result of limited durable response rates, cumulative viral resistance, and/or poor tolerability. Telbivudine has potent antiviral activity against hepatitis B virus (HBV) in vitro and in the woodchuck model and has a promising preclinical safety profile. In this first clinical study of telbivudine, safety, antiviral activity, and pharmacokinetics were assessed in 43 adults with hepatitis B e antigen-positive chronic hepatitis B. This placebo-controlled dose-escalation trial investigated 6 telbivudine daily dosing levels (25, 50, 100, 200, 400, and 800 mg/d); treatment was given for 4 weeks, with 12 weeks' follow-up. Serum HBV DNA levels were monitored via quantitative polymerase chain reaction. The results indicate that telbivudine was well tolerated at all dosing levels, with no dose-related or treatment-related clinical or laboratory adverse events. telbivudine plasma pharmacokinetics were dose-proportional within the studied dose range. Marked dose-related antiviral activity was evident, with a maximum at telbivudine doses of 400 mg/d or more. In the 800 mg/d cohort, the mean HBV DNA reduction was 3.75 log10 copies/mL at week 4, comprising a 99.98% reduction in serum viral load. Correspondingly, posttreatment return of viral load was slowest in the high-dose groups. Viral dynamic analyses suggested a high degree of efficiency of inhibition of HBV replication by telbivudine and helped refine selection of the optimal dose. In conclusion, these results support expanded clinical studies of this new agent for the treatment of hepatitis B. PMID- 15349913 TI - Quantitation of covalently closed circular hepatitis B virus DNA in chronic hepatitis B patients. AB - This study examined a signal amplification assay, the Invader assay, for the quantitation of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) in liver biopsies and sera. DNA was extracted from liver biopsy and serum samples were collected from 16 hepatitis B e antigen (HBeAg)-positive and 36 antibody-to HBeAg-positive (anti-HBe-positive) chronic hepatitis B patients. The amount of total HBV DNA and cccDNA was measured using the Invader assay. Anti-HBe-positive patients had lower median total intrahepatic HBV DNA (P < .001) and intrahepatic cccDNA levels (P = .001) than HBeAg-positive patients. Intrahepatic cccDNA correlated positively with the total intrahepatic HBV DNA (r = 0.950, P < .001). However, the proportion of intrahepatic HBV DNA in the form of cccDNA was inversely related to the amount of total intrahepatic HBV DNA (r = -0.822, P < .001). A small amount of cccDNA was detected in 39 of 52 (75%) serum samples. Anti-HBe-positive patients had lower median serum cccDNA levels than HBeAg positive patients (P = .002). Serum HBV DNA correlated positively with intrahepatic total HBV DNA (r = 0.778, P < .001) and intrahepatic cccDNA (r = 0.481, P = .002). In conclusion, the Invader assay is a reliable assay for the quantitation of cccDNA. Serum and intrahepatic total HBV DNA and cccDNA levels become lower as the disease progresses from HBeAg-positive to anti-HBe-positive phase, with cccDNA becoming the predominant form of intrahepatic HBV DNA. PMID- 15349914 TI - Functional impairment of myeloid and plasmacytoid dendritic cells of patients with chronic hepatitis B. AB - Dendritic cells (DC) play an important role in the induction of T-cell responses. We hypothesize that the hampered antiviral T-cell response in chronic hepatitis B patients is a result of impaired dendritic cell function. In this study, we compared the number, phenotype and functionality of two important blood precursor DC, myeloid DC (mDC) and plasmacytoid DC (pDC), of chronic hepatitis B patients with healthy volunteers. No differences in percentages of mDC and pDC in peripheral blood mononuclear cells were observed between chronic hepatitis B patients and healthy controls. The allostimulatory capacity of isolated and in vitro matured mDC, but not of pDC, was significantly decreased in patients compared to controls. Accordingly, a decreased percentage of mDC expressing CD80 and CD86 was observed after maturation, compared to controls. In addition, mDC of patients showed a reduced capacity to produce tumor necrosis factor alpha after a stimulus with synthetic double-stranded RNA and interferon gamma. Purified pDC from patients produced less interferon alpha, an important antiviral cytokine, in response to stimulation with Staphylococcus aureus Cowan strain I than pDC isolated from controls. In conclusion, mDC and pDC are functionally impaired in patients with chronic hepatitis B. This might be an important way by which hepatitis B virus evades an adequate immune response, leading to viral persistence and disease chronicity. PMID- 15349915 TI - A case-control study for differences among hepatitis B virus infections of genotypes A (subtypes Aa and Ae) and D. AB - There are two subtypes of hepatitis B virus genotype A (HBV/A) and they are provisionally designated Aa ("a" standing for Africa/Asia) and Ae ("e" for Europe). In a case-control study, 78 HBV/Aa, 78HBV/Ae, and 78HBV/D carriers from several countries were compared. The prevalence of HBe antigen (HBeAg) in serum was significantly lower in carriers of HBV/Aa than in carriers of HBV/Ae (31% vs. 49%; P = .033), with a difference more obvious in the carriers aged 30 years or younger (34% vs. 67%; P = .029). HBV DNA levels in the carriers of HBV/Aa (median, 3.46 log copies/mL; 95% CI, 2.93-3.95) were significantly lower than those of carriers of HBV/Ae (6.09 log copies/mL; 95% CI, 4.24-7.64) or of carriers of HBV/D (5.48 log copies/mL; 95% CI, 4.06-7.02), regardless of the HBeAg status (P < .001). The most specific and frequent substitutions in 54 HBV/Aa isolates were double substitutions for T1809 (100%) and T1812 (96%) immediately upstream of the precore initiation codon, which would interfere with the translation of HBeAg in HBV/Aa infections. They were not detected in 57 HBV/Ae or 61 HBV/D isolates examined. The double mutation in the core promoter (T1762/A1764) was more frequent in both HBV/Aa (50%) and HBV/Ae (44%) than in HBV/D isolates (25%; P < .01), whereas the precore mutation (A1896) occurred in HBV/D isolates only (48%; P < .0001). In conclusion, the clearance of HBeAg from serum may occur by different mechanisms in HBV/Aa, HBV/Ae, and HBV/D infections, which may influence clinical manifestations in the Western countries where both genotypes A and D are prevalent. PMID- 15349916 TI - Reexamining the role of the humoral immune response in control of hepatitis C virus infection. PMID- 15349917 TI - Genetic manipulations utilizing albumin and alpha-fetoprotein promoter/enhancers affect both hepatocytes and oval cells. PMID- 15349918 TI - Peginterferon alfa-2a (40 kd) and ribavirin for black American patients with chronic HCV genotype 1. PMID- 15349921 TI - No hepatic iron overload 12 years after liver transplantation for hereditary hemochromatosis. PMID- 15349923 TI - The legend of the lardaceous liver. PMID- 15349927 TI - Separation and recovery of intact gold-virus complex by agarose electrophoresis and electroelution: application to the purification of cowpea mosaic virus and colloidal gold complex. AB - Colloidal gold has been coupled to a mutant cowpea mosaic virus (CPMV), which contains 60 cysteine residues on the surface. A purification process was developed to separate the gold-containing viral nanoblocks (VNBs) from the free gold. Agarose electrophoresis was utilized to separate the mixture followed by electroelution of the desired sample to recover the intact virus. Mobility of Au VNB and free colloidal gold was facilitated by the addition of thioctic acid (TA). 30% of the gold-containing virus was recovered after electroelution as determined by absorbance measurements. Histogram analysis of transmission electron microscopy (TEM) images demonstrated the efficient separation of gold containing virus from free gold. TEM and scanning electron microscopy (SEM) images indicated that the virus was recovered intact. Monodisperse spherical particles of nominal size of 45 nm were observed under SEM. PMID- 15349928 TI - Determination of pore/protein size via electrophoresis and slit sieve model. AB - We used electrophoresis for three purposes: (i) estimation of the mean pore size of polyacrylamide gels via measuring electrophoretic mobility of globular proteins of known sizes in combination with simple sieve (cylindrical and slit) models; (ii) determination of the average size of protein molecules (native or denatured) by the use of the same models; (iii) monitoring the changes in molecular dimensions of proteins in the course of their denaturation. Both models yield results that are in good agreement with those found via the more elaborate techniques (considering the principal differences involved). The approach provides a direct and convenient way of monitoring the variations in protein sizes during the course of their denaturation in gels having a gradient of denaturants, and possibly the number of conformational states involved in the process, a facet that is quite unique and useful. The simpler slit model seems to yield better results in the latter case and is moreover supported by the recently reported data on electrophoresis of DNA molecules through the 1 microm slits of a microbrush matrix made of micropillars arranged in a hexagonal lattice. PMID- 15349929 TI - Cerebrospinal fluid amyloid beta peptide patterns in Alzheimer's disease patients and nondemented controls depend on sample pretreatment: indication of carrier mediated epitope masking of amyloid beta peptides. AB - A quantitative urea-based amyloid beta (Abeta)-sodium dodecyl sulfate polyacrylamide gel electrophoresis with Western immunoblot (Abeta-SDS PAGE/immunoblot) reveals highly conserved and disease-specific Abeta peptide patterns (Abeta 1-37, 1-38, 1-39, 1-40, 1-42) in Alzheimer's disease (AD) patients and nondemented controls. For further standardization of this method, we analyzed cerebrospinal fluid (CSF) of eight probable AD patients and seven nondemented controls using different preanalytical procedures for Abeta-SDS PAGE/immunoblot and Abeta1-42-enzyme linked immunosorbent assay (ELISA). Both diagnostic groups were discriminated significantly by absolute levels of Abeta1 42 and ratios of Abeta1-42/40, 1-42/38, 1-42/39. Preanalytical freezing of CSF led to a highly significant loss of all Abeta peptide species. This effect was most pronounced for Abeta1-42 and completely prevented by SDS-heat denaturation prior to freezing. Prolonged storage of SDS-heat denatured CSF led to a selective loss of Abeta1-42 and impaired the discrimination of diagnostic groups as measured by Abeta-SDS-PAGE/immunoblot. Neither freezing nor storage significantly affected absolute Abeta1-42 levels as determined by Abeta1-42-ELISA, but both impaired the discrimination of diagnostic groups. Hence, we suggest immediate analysis of samples for Abeta1-42-ELISA, analysis after a short freezing interval for Abeta-SDS-PAGE/immunoblot, and avoidance of prolonged storage intervals. Remarkably, Abeta-SDS-PAGE/immunoblot measured threefold higher levels of Abeta1 42 in CSF than Abeta1-42-ELISA. In summary, our results indicate carrier-mediated epitope masking of Abeta1-42. PMID- 15349930 TI - Polyacrylamide gel electrophoresis of fluorophore-labeled hyaluronan and chondroitin sulfate disaccharides: application to the analysis in cells and tissues. AB - This report describes a new formulation of polyacrylamide gel electrophoresis of fluorophore-labeled saccharides (PAGEFS) for the analysis of hyaluronan (HA) and chondroitin sulfate (CS) Delta-disaccharides. PAGEFS relies on derivatization of reducing ends of HA- and the variously sulfated CS-derived Delta-disaccharides with 2-aminoacridone (AMAC), followed by electrophoresis under optimized buffer conditions (Tris-borate and Tris-HCl) and on polyacrylamide gels (25% T/3.75% C). The method was applied to the analysis of glycosaminoglycans (GAGs) from the human umbilical cord tissue and GAGs isolated from human aortic smooth muscle cell cultures. The obtained results were in agreement with those obtained after an analysis with high-performance liquid chromatography (HPLC). On the basis of these results, PAGEFS is a rapid and sensitive method for the analysis of the total amount of HA- and CS-derived disaccharides, as it allows analyzing 20 samples in minigels in one run and provides quantitation with relatively high sensitivity (less than 25 pmol per disaccharide). In addition, PAGEFS overcomes the lack of commercial gels described previously for the separation of AMAC labeled disaccharides. Therefore, the method proposed here is an economic and useful tool for a fast screening of GAGs in biological samples, particularly when a high number of samples should be analyzed. PMID- 15349931 TI - Detection of glutathione reductase after electrophoresis on native or sodium dodecyl sulfate polyacrylamide gels. AB - Commercial glutathione reductase (GR) from spinach and yeast (Saccharomyces cerevisiae) were stained on 7.5% native polyacrylamide gel electrophoresis (PAGE) gels or 15% sodium dodecyl sulfate (SDS)-PAGE gels with or without further purification by a 2',5'-ADP Sepharose 4B affinity column. For SDS-PAGE gels, the SDS was removed first by washing twice with 25% isopropanol in 10 mM Tris-HCl (pH 7.9) for 10 min. The gel was then dipped in a 50 mM Tris-HCl buffer (pH 7.9) containing 4.0 mM oxidized glutathione (GSSG), 1.5 mM NADPH, and 2 mM 5,5' dithiobis(2-nitrobenzoic acid) (DTNB) for 20 min. The GR activity was negatively stained in the dark by a solution containing 1.2 mM 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) and 1.6 mM phenazine methosulfate (PMS) for 5-10 min. The contrast between the clear zone of GR activity and the purple background was found in both native and SDS-PAGE gels. This negative staining method can detect GR as little as 0.064 units and 0.0032 units, respectively, for spinach and yeast sources. Under reduced SDS-PAGE gels, the GR activity band located on 72 kDa for spinach and 51 kDa for yeast. This fast and sensitive method could be used during enzyme purification and for characterization of GR from different sources under different physiological stages or conditions. PMID- 15349932 TI - Improved resolution power of electrophoretic fractionation of DNA using a voltage gradient "up and down" application. AB - The improved resolution power of electrophoretic fractionation of DNA in a wide range of molecular masses is demonstrated using an "up and down" application of voltage gradient gel electrophoresis (VGGE). This application also allows separation of different DNA fragments which are poorly fractionated in conventional electrophoresis. PMID- 15349933 TI - Nonlinear electroosmosis in hierarchical monolithic structures. AB - We studied the dependence of electroosmotic flow (EOF) velocity and separation efficiency for neutral analytes in 100 microm ID capillary monoliths on a variation of the mobile phase ionic strength and applied electrical field strength, i.e., we covered a range for the concentration of Tris buffer from 10( 5) to 10(-2) M and applied electrical field strengths up to 10(5) V/m. The silica based monoliths are hierarchically structured having intraskeleton mesopores and interskeleton macropores. While a linear dependence of the average EOF velocity on applied field strength could be observed with 5 x 10(-3) M Tris (turning slightly nonlinear at a higher concentration due to thermal effects), this dependence becomes systematically nonlinear as the Tris concentration is reduced towards 10(-4) M. Increased velocities by more than 50% compared to those expected from linear behavior are realized at 10(5) V/m. Concomitantly, as the Tris concentration is reduced from 10(-3) to 10(-4) M, we notice an improvement in plate heights by a factor of more than 2 (they approach 2 microm for ethylbenzoate). We complementary analyzed the onset of the nonlinear EOF dynamics in a hierarchical monolith and the significantly reduced axial dispersion in view of nonequilibrium electrokinetic effects which may develop in porous media due to the presence of ion-permselective regions, e.g., the mesoporous monolith skeleton. In this respect, a decreasing mobile phase ionic strength favors the formation of nonequilibrium concentration polarization in strong electrical fields, and a coupling of the electrostatics and hydrodynamics then may explain nonlinear EOF velocities and increasing separation efficiencies depending on the Tris concentration and applied field strength. PMID- 15349934 TI - Formamide as solvent for capillary zone electrophoresis. AB - A comprehensive investigation of a number of aspects when using formamide as background electrolyte solvent in capillary zone electrophoresis was presented. It included (i) the change of the ion mobility with ionic strength, (ii) the influence of the ionic strength on diffusion coefficients, and (iii) on the separation efficiency expressed by the maximum reachable plate numbers (when only longitudinal diffusion contributed to zone broadening), (iv) the effect of the solvent on pKa values (taken from the literature) of neutral and cation acids, (v) the establishment of the a pH scale in formamide by dissolving acids with known pKa values and their salts at defined proportion (thus circumventing the problem of calibrating the pH meter), (vi) the agreement between the experimentally derived and the theoretical dependence of the effective mobility on pH, (vii) the uptake of water of this hygroscopic solvent from the humidity of the environment and its consequence to the ion mobilities, pKa values, and the chemical stability of the solvent (e.g., hydrolysis), and finally (viii) the use of conductivity and indirect UV absorption to enable detection of analytes below the optical cutoff of formamide. PMID- 15349935 TI - Capillary isoelectric focusing--reproducibility and protein adsorption. AB - Capillary isoelectric focusing (CIEF) is an important tool for the quality assurance of biotechnologically maintained drugs and for proteome analysis. The critical performance parameters of this technique are the precisions of isoelectric point (pI) values and peak areas. Compared to capillary zone electrophoresis (CZE), where precise results can be obtained (e.g., 0.5% relative standard deviation (RSD) for peak areas, n = 60), only few data are available for CIEF experiments. So far, reproducible data of pI values (RSD = 0.5%) have been acquired, but peak areas show inferior results (about 3-15% RSD). Nonstable capillary coatings and protein adsorption have been discussed as possible reasons. Recent work of Righetti et al. [25, 27] has proven that the use of coated capillaries can reduce the adsorption of proteins by 50% but cannot prevent it. In our CIEF experiments irregular and poorly reproducible peak patterns have been observed. In a long-time experiment of 106 repeated runs, an overall RSD of 10% was obtained for peak areas, RSD of 2% only in series of about 10 consecutive replicates. Especially at higher concentrations the reproducibility deteriorates. This seems to be the result of a self-amplifying process, induced by adsorbed protein molecules, leading to further agglomerations. CZE control experiments in linear polyacrylamide (LPA)-coated capillaries proved a strong pH dependency of these effects within a small range. Compared to bare fused-silica surfaces, adsorption effects are reduced but not inhibited. An enhancement of reproducibility in CIEF experiments can be achieved only by controlling the interactions of proteins and capillary walls. PMID- 15349936 TI - Disposable twin gold electrodes for amperometric detection in capillary electrophoresis. AB - We describe a simple and easy way to construct gold microelectrodes for amperometric detection in capillary electrophoresis (CE). The gold microelectrodes, in single or twin sets, were obtained from recordable compact discs (gold-sputtered type), which present highly reproducible surface characteristics. The performance of these electrodes was evaluated by using a home-made CE equipment. The basic steps for the electrode construction are: drawing on a microcomputer; laser printing of the design on wax paper; heat transfer of the toner onto the gold surface of a peeled recordable compact disc (CD-R); etching of the gold layer from unprinted regions; removal of the toner with a solvent; sealing of unused electrode areas with varnish. One electrode at a time was connected to a potentiostat (or two, to a bipotentiostat) and operated in a wall jet configuration relative to the CE capillary outlet. The amperometric signals were integrated for quantification purposes. Repetitive injections (n = 10) of a mixture containing iodide, ascorbic acid, dipyrone, and acetaminophen (20, 200, 500, and 100 microM), presented relative standard deviations of 2.9, 4.5, 6.1, and 4.0%, respectively. For these analytes, the detection limits (S/N = 3, 30 s of 100 mm hydrodynamic injection) were 0.1, 0.5, 3.1, and 1.1 microM, respectively. PMID- 15349937 TI - Prediction of wheat dough W and P/L inflation test parameters by capillary zone electrophoresis of a protein extract followed by multivariate regression. AB - A procedure for the evaluation of the wheat flour hardness, based on capillary electrophoresis of a protein extract in an isoelectric acidic buffer, was developed. The 13 flour samples were extracted twice, and two injections of each extract were made. Separations were performed in a background electrolyte (BGE) containing 40 mM aspartic acid, 6 M urea, and 0.5% hydroxyethylcellulose at 60 degrees C. Using the normalized and corrected areas of 79 peaks and peak groups, a partial least squares regression (PLS1) model was able to predict the flour strength or dough deformation work (W) and the dough tenacity/extensibility ratio (P/L) (Alveograph parameters) with an average relative standard deviation in the predictions of +/- 3% and +/-8%, respectively. These values amounted to a +/- 6 8% and +/- 11% with multiple linear regression (MLR) and PLS1 models constructed by measuring only 12 peaks and peak group areas on the electropherograms. PMID- 15349938 TI - Determination of amino acids in rat vitreous perfusates by capillary electrophoresis. AB - In vivo determinations of amino acids are important for improving our understanding of physiological states of biological tissue function and dysfunction. However, the chemically complex matrix of different biological fluids complicates the assay of this important class of molecules. We introduce a method for characterizing the amino acid composition of submicroliter volumes of vitreous humor perfusates. Low-flow push-pull perfusion sampling is compatible with collecting small volume samples in a complicated matrix that are potentially difficult to separate. An efficient, sensitive, and rapid analysis of amino acids from in vivo perfusates of the vitreous is presented with 3-(4-carboxybenzoyl)-2 quinoline-carboxaldehyde (CBQCA) derivatitation and capillary electrophoresis (CE) separation with laser-induced fluorescence detection (LIF). Derivatization with CBQCA for up to 2 h provided high sensitivity and low detection limits at the nM level. Seventeen amino acids including D-serine (D-Ser) and D-aspartate (D Asp) were resolved in less than 10 min. Importantly, D-Ser is separated from its enantiomeric pair. Characterization of vitreal amino acids with this assay technique will be useful for understanding ocular diseases and physiological mechanisms in vision. PMID- 15349939 TI - Charge density profiling of circulating human low-density lipoprotein particles by capillary zone electrophoresis. AB - Capillary zone electrophoresis (CZE) has been utilized to profile the low-density (LDL) particles in human blood serum in this study. A 5 mM sodium phosphate buffer, pH 7.40, was chosen as the most suitable CE buffer and an extensive ultrafiltration (UF) procedure was applied to purify the LDL sample. Two LDL particle species, LDL with lower mobility and LDL- with higher mobility were observed. The electropherograms were highly reproducible with good precision of effective mobilities, corrected peak areas (CPAs) and CPA ratio of LDL-/LDL. LDL particles shown on the electropherogram were also characterized by several procedures. The applications of Sigma HDL cholesterol reagent and CE on-line 2 propanol precipitation indicated that the two particle species shown in the electropherogram belong to LDL. The LDL particles were found to associate with the buoyant LDL fraction and the LDL- particles associate with the dense LDL fraction. This study utilizes CZE for the profiling of LDL isoforms and provides a new analytical method for the resolution of LDL subspecies. It demonstrates a high-mobility LDL particle which circulates in healthy subjects and diminishes in atherosclerotic patients. Diminution of the high-mobility LDL subspecies may be linked to minimal formation of arterial plaque in atherosclerotic patients. PMID- 15349940 TI - In situ monitoring of kinetics of metabolic conversion of ATP to ADP catalyzed by MgATPases of muscle Gastrocnemius skinned fibers using micellar electrokinetic chromatography. AB - A method for the in situ measurement of the kinetics of ATP metabolic transformation using capillary electrophoresis (CE) has been developed. The depletion of ATP and formation of ADP were monitored in situ by using saponin permeabilized muscle fibers. The method of micellar electrokinetic chromatography, employing reversed electroosmotic flow by cationic surfactant and reversed-polarity mode, provided an efficient and reproducible separation of nucleotides and enabled kinetic analysis of the reaction to be performed in a large range of nucleotide concentrations that approaches physiological concentrations of ATP in the muscle cells, without the need for precipitation of proteins prior to sample application. The analytes were detected at a nM level with a reproducibility of about 7%. This reproducibility enabled the comparison of different competing kinetic models of ATP conversion to ADP and the results show that the MgATPase activity in the fast-twitch gastrocnemius muscle followed biphasic kinetics that corresponds to the allosteric character of regulation of the enzyme(s) activity at physiological ATP concentrations. The results also confirmed that the combination of minimal sample volume requirements, rapid measurement and reproducibility makes the micellar CE a valuable tool for the analysis of biological fluids and understanding the processes of biological interest. PMID- 15349941 TI - Separation and determination of aconitine alkaloids in traditional Chinese herbs by nonaqueous capillary electrophoresis. AB - An easy, rapid, and simple nonaqueous capillary electrophoresis (NACE) method was developed for the identification and determination of three aconitine alkaloids, hypaconitine (HN), aconitine (AN), and mesaconitine (MN) within 6 min. The most suitable running buffer was composed of 60 mM ammonium acetate, 0.5% acetic acid, and 15% acetonitrile (ACN) in methanol with a fused-silica capillary column (50 cm x 75 microm ID). In the concentration range 12.5-1000 mg/L the calibration curves reveal linear relationships between the peak area for each analyte and its concentration (correlation coefficients: 0.9997 for HN, 0.9999 for AN, and 0.9995 for MN). The relative standard deviations of the migration time and peak area of the three alkaloids were 0.13, 0.57, 0.33 and 2.87, 1.06, 3.49%, respectively. The method was successfully applied to determine the three alkaloids in two commonly used traditional Chinese herbal medicines, the recoveries of the three constituents ranging between 94.7-101.9% for HN, 98.3-102.3% for AN, and 98.1 104.6% for MN. PMID- 15349942 TI - Capillary and microchip electrophoresis of basic drugs with contactless conductivity detection. AB - The extension of contactless conductivity detection in electrophoresis to the determination of basic drugs is demonstrated using beta-adrenergic blocking agents (beta-blockers) and other physiologically active amines as examples. The high-voltage approach to conductivity detection was employed for conventional capillaries as well as microchip devices. Acidic buffers were used in all cases. A buffer consisting of 100 mM acetic acid and 1 mM histidine was deemed most optimal for the separation of six beta-blockers and best results for the analysis of the other amines were achieved with a 20 mM lactic acid buffer at low pH value. The detection limits ranged from 0.06 to 5 microM. To demonstrate potential practical applications, a main component assay was conducted for three pharmaceutical formulations. On-chip, five pharmaceutical amines could be baseline-resolved in a 8 cm long microchannel in 90 s, albeit a reduced sensitivity and peak capacity compared to conventional capillary electrophoresis. PMID- 15349943 TI - Microchip capillary electrophoresis with a boron-doped diamond electrochemical detector for analysis of aromatic amines. AB - The attractive features of a boron-doped diamond (BDD) thin-film detector for microchip capillary electrophoretic (CE) separations of dye-related amino substituted aromatic compounds are described. The diamond electrode was employed in the end-column amperometric detection of 4-aminophenol (4-AP), 1,2 phenylenediamine (1,2-PDA), 2-aminonaphthalene (2-AN), 2-chloroaniline (2-CA), and o-aminobenzoic acid (o-ABA), and its attractive behavior was compared to commonly used screen-printed carbon and glassy-carbon electrodes. These conventional electrode materials exhibit a significant degree of passivation and low sensitivity to the above-mentioned environmental pollutants. The diamond based electrochemical detection system displayed a favorable analytical performance, including lower noise levels, higher peak resolution with enhanced sensitivity, and improved resistance against electrode passivation. Factors influencing the on-chip analysis were assessed and optimized. The diamond detector displayed detection limits of 2.0 and 1.3 microM for 4-AP and 2-AN, respectively, and a wide linear response for these compounds over the 2-50 microM range. The enhanced stability was demonstrated by relative standard deviation (RSD) values of 1.4% and 4.7% for 100 microM 1,2-PDA and 200 microM 2-CA, respectively, for repetitive detections (n = 7). Besides, the simultaneously observed current decrease was 2.4 and 9.1% for 1,2-PDA and 2-CA, respectively (compared to 21.8 and 41.0% at the screen-printed carbon electrode and 28.3 and 34.1% at the glassy carbon electrode, respectively). The favorable properties of the diamond electrode indicate great promise for environmental applications in CE and other microchip devices. PMID- 15349944 TI - Separation of proteins on surface-modified poly(dimethylsiloxane) microfluidic devices. AB - Separation and detection of proteins have been realized on nonionic surfactant modified poly(dimethylsiloxane) (PDMS) microfabricated devices with end-column amperometric detection. The hydrophobic PDMS channels are turned into hydrophilic ones after being modified with Brij35 and facilitate the separation of proteins. The coating can remarkably reduce the adsorption of large protein molecules and is stable in the range of pH 6-12. The detection of proteins in such channels needs less rinsing time and thus efficiency is raised. Even large molecules of proteins can also be detected with better reproducibility and enhanced plate numbers. The relative standard deviation (RSD) of the migration time for glucose oxidase (GOD) is 2.2% (n = 19). Separation of GOD and myoglobin has been developed in modified channels. Predominant operational variables, such as the coating conditions, the concentration of surfactant and buffer, are studied in detail. PMID- 15349945 TI - Determination of SARS-coronavirus by a microfluidic chip system. AB - We have developed a new experimental system based on a microfluidic chip to determine severe acute respiratory syndrome coronavirus (SARS-CoV). The system includes a laser-induced fluorescence microfluidic chip analyzer, a glass microchip for both polymerase chain reaction (PCR) and capillary electrophoresis, a chip thermal cycler based on dual Peltier thermoelectric elements, a reverse transcription-polymerase chain reaction (RT-PCR) SARS diagnostic kit, and a DNA electrophoretic sizing kit. The system allows efficient cDNA amplification of SARS-CoV followed by electrophoretic sizing and detection on the same chip. To enhance the reliability of RT-PCR on SARS-CoV detection, duplex PCR was developed on the microchip. The assay was carried out on a home-made microfluidic chip system. The positive and the negative control were cDNA fragments of SARS-CoV and parainfluenza virus, respectively. The test results showed that 17 positive samples were obtained among 18 samples of nasopharyngeal swabs from clinically diagnosed SARS patients. However, 12 positive results from the same 18 samples were obtained by the conventional RT-PCR with agarose gel electrophoresis detection. The SARS virus species can be analyzed with high positive rate and rapidity on the microfluidic chip system. PMID- 15349946 TI - High reproducibility of large-gel two-dimensional electrophoresis. AB - Two-dimensional gel electrophoresis (2-DE) facilitates the separation of thousands of proteins from highly complex protein mixtures and has become a central method in proteomics in recent years. In the present study, we examined the technical variability of large 2-DE gels with respect to sample preparation, electrophoresis procedure, data acquisition, and biological variation by analyzing a disease (Huntington's disease) and control state with a commercially available software package, PROTEOMWEAVER trade mark. Scatter plots and correlation coefficients were obtained to quantify both technical and biological variation. Even 2-DE gels run separately in both dimensions yielded correlation coefficients around 0.88 and deviations from the mean close to 20% for low intensity spots. This indicates a high technical reproducibility of the 2-DE procedure developed in our laboratory. Variability within a biological condition was low and comparable to technical variation (at least 0.87). Two-dimensional (2 D) gels obtained from samples of different biological conditions (health vs. disease) achieved a variability similar to intracondition and technical variability. These findings highlight the importance of multiple gel and spot-by spot comparisons to identify biological significant changes. Minor errors introduced by technical and biological variation allow a comparison of all gels within a study which facilitates the tackling of complex biological problems. PMID- 15349947 TI - A statistical comparison of silver and SYPRO Ruby staining for proteomic analysis. AB - Silver staining has been the method most commonly employed for high sensitivity staining of proteins following two-dimensional gel electrophoresis. Whilst this method offers detection in the nanogram range it does have major drawbacks including a lack of linearity, nonstoichiometric staining of proteins, a lack of compatibility with the microchemical preparation of proteins for identification by mass spectrometric techniques, and a highly subjective assessment of the staining endpoint. SYPRO Ruby is a relatively new, ruthenium complex-based stain which is reported to offer advantages over silver, particularly in overcoming the limitations cited above. We describe a series of experiments where several protein staining procedures commonly employed are compared. To enable optimization of the in situ digestion procedure, a statistical approach has been undertaken. The effects of a variety of staining, digestion, and analysis protocols on the downstream processing of a test radiolabeled protein were studied. The data confirms that as well as offering sensitivity similar to silver, SYPRO Ruby staining is reproducible, linear, and offers a higher level of compatibility with the identification of proteins by mass spectrometry. PMID- 15349948 TI - A mouse serum two-dimensional gel map: application to profiling burn injury and infection. AB - With the importance of mouse as a model to study human diseases and the human and rat plasma/serum two-dimensional (2-D) maps being extensively annotated, this study was aimed at constructing a detailed mouse serum 2-D map. Serum proteins from two different inbred strains of mice (BALB/cJ and C57BL/6J) and mice subjected to two different inflammatory stimuli (20% burn injury and lipopolysaccharide (LPS) injection) were separated on overlapping gels covering pH 3-8 and stained with SYPRO Ruby dye. The tryptic peptides from the resolved spots were analyzed by mass spectrometry, leading to the identification of 38 different gene products. With the exception of major urinary proteins found in abundance in male C57BL/6J mice, little strain difference of the mouse serum 2-D was observed. Many proteins detected in the mouse serum 2-D map were not reported in human or rat serum 2-D maps including epidermal growth factor receptor. Three major murine acute-phase proteins (APPs), haptoglobin, serum amyloid A, and serum amyloid P, were highly induced by both inflammatory stimuli. Image analysis shows that the variations of APPs between these two inflammatory models were not uniform although LPS (100 microg/animal) in general was more effective than 20% burn injury in inducing APPs. Serum amyloid A, much more sensitive to endotoxin than burn injury, may represent a sensitive marker to differentiate these two different inflammatory states. PMID- 15349951 TI - Novel method for the determination of solubility in aerosol propellants. AB - A new on-line reverse-phase high-performance liquid chromatography method to determine the solubility of compounds in propellant-based metered-dose inhaler (MDI) formulations was developed. The new method uses a direct injection from an MDI vial into the port of a manual injector. The MDI vials were coupled with a filtration and injection assembly to filter the excess compound, and deliver the filtrate of the MDI vials to the injector port of the manual injector. A backpressure regulator was connected to the manual injector, to maintain the propellant in the liquid state after injection from the MDI vial. Phase separation studies were conducted to investigate the miscibility of hydrofluoroalkane 134a with different mobile-phase solvent compositions. Characterization of the new direct injection method was done by evaluating sample presentation and chromatography variables to determine the robust nature of the method. Beclomethasone dipropionate in ethanol/hydrofluoroalkane 134a was used as a model system for development. A chemically diverse set of nine compounds was used to evaluate solubility measurement reproducibility. Solubilities were determined at two different temperatures (25 and 37 degrees C) and the average relative standard deviation for all of the solubility measurements was <4%. PMID- 15349952 TI - Release-modulating factors strongly affecting steroid diffusion from silicone elastomer. AB - Investigations were undertaken to determine the cause of decreases over time in the release rate from levonorgestrel (LNG) implants consisting of silicone elastomer tubing filled with crystalline steroid. Emptying and refilling with the same steroid partially restored release rate. Surprisingly, a further increment in release rates was attained if the tubing was briefly irrigated with methanol before refill. Fractional crystallization showed that release-modulating factors could be concentrated in mother liquors and were initially present as impurities. Boiling LNG in ethanol or methanol produced a number of release-modulating factors of which the most prominent was also found in one production lot of LNG. It was identified as 6beta-hydroxy-levonorgestrel (6beta-OH-LNG). Added to LNG at the 2% level, 6beta-OH-LNG decreased the release rate by 27%. PMID- 15349953 TI - Effect of chemical enhancers and iontophoresis on thiocolchicoside permeation across rabbit and human skin in vitro. AB - The aim of this work was to study the permeation of thiocolchicoside across the skin in vitro. The effect of the chemical enhancer lauric acid and the physical technique of iontophoresis was investigated. Permeation experiments were performed in vitro using rabbit ear skin as barrier. The effect of lauric acid at different concentrations (2% and 4%) and of the vehicle (water, ethanol, or ethanol/water) was investigated. The primary effect of lauric acid was on the partitioning parameter, whereas the diffusive parameter did not change significantly. When human epidermis was used, the permeation parameters were generally lower, although not significantly different from rabbit ear skin. The data obtained with full-thickness human skin indicate that, despite the hydrophilic nature of thiocolchicoside, the resistance to drug transport is not limited to the stratum corneum, but that the underlying dermal tissue can also contribute. Iontophoresis enhanced the flux of thiocolchicoside compared with the passive control. The mechanism by which iontophoresis enhanced thiocolchicoside transport across the skin was electroosmosis. The permeation of thiocolchicoside across the skin can be enhanced using chemical or physical penetration enhancers. PMID- 15349954 TI - Quantification of glycine crystallinity by near-infrared (NIR) spectroscopy. AB - The object of this investigation was to use near-infrared (NIR) spectroscopy for quantification of glycine crystallinity. Glycine samples, with different degrees of crystallinity, were obtained by physically mixing different proportions of crystalline beta-glycine with amorphous glycine. NIR spectra were obtained, directly from samples in glass vials, over the wavelength range of 1100-2500 nm. A partial least squares (PLS) model was developed to correlate the NIR spectral changes with the degree of crystallinity. Using this model, a standard error of calibration (SEC) of 2.1% was obtained with an r(2) value of 0.996. Cross validation was used to test the precision of the quantitative model, resulting in a standard error of prediction (SEP) of 3.2%. These results indicate that NIR spectroscopy is well suited to the measurement of glycine crystallinity in lyophilized products. Employing the PLS model, the crystallinity of glycine in freeze-dried sucrose-glycine mixtures was evaluated. At a sucrose to glycine ratio >4, glycine crystallization during lyophilization was inhibited. Conversely, at ratios < or =0.67, glycine remained substantially crystalline. At intermediate compositions, the glycine was partially crystalline. PMID- 15349955 TI - Resveratrol glucuronides as the metabolites of resveratrol in humans: characterization, synthesis, and anti-HIV activity. AB - Resveratrol is a natural product with diverse biological activities. We have previously reported that resveratrol possesses potent synergistic inhibitory activity against human immunodeficiency virus (HIV)-1 infection in combination with nucleoside analogs (Heredia et al. 2000. J Acquir Immune Defic Syndr 25:246 255). As a part of our program in developing resveratrol as a component for anti HIV chemotherapy, we describe in this article the characterization, chemical synthesis, and biological effects of the human metabolites of resveratrol. We found that resveratrol was metabolized in humans into two metabolites, which were characterized as resveratrol-3-O- and 4'-O-glucuronides. For further biological studies, we reported two simple, alternative methods for the synthesis of the metabolites. The cytotoxic and antiviral activities of resveratrol and its metabolites were compared in cell culture experiments using human peripheral blood mononuclear cells. Whereas resveratrol was cytotoxic at > or =30 microM, no cytotoxicity was observed for the metabolites at concentrations as high as 300 microM. However, resveratrol showed strong synergistic anti-HIV activity with didanosine at 10 microM, but no synergistic effects were observed for either of the metabolites at up to 300 microM. Nevertheless, the in vitro activity of the metabolites (resveratrol glucuronides) may not necessarily reflect their in vivo function, given the fact that the ubiquitously existing human beta-glucuronidase could convert the metabolites back to resveratrol locally or systematically in vivo. The present studies have implications for future development of resveratrol and/or its derivatives as a chemotherapeutic agent. PMID- 15349956 TI - Development of stable liquid formulations for adenovirus-based vaccines. AB - We have evaluated the stability profiles of adenovirus type-5 (Ad5)-based vaccine formulations to identify liquid formulations that are stable during long-term storage at 4 degrees C. By identifying the major physiochemical inactivation pathway(s) during storage, formulations of Ad5 were designed with specific pharmaceutical excipients leading to greatly enhanced stability. For example, results indicate that Ad5 is stabilized by non-ionic surfactants and cryoprotectants as well as excipients known to inhibit free-radical oxidation. A non-ionic surfactant is necessary to prevent adsorption of adenovirus to glass surfaces during storage, and a cryoprotectant is needed to prevent freeze-thaw induced virus inactivation. In a base formulation (A105) containing sucrose as the cryoprotectant and polysorbate-80 as the non-ionic surfactant, metal-ion catalyzed free-radical oxidation is an important mechanism of Ad5 inactivation. The free-radical oxidation inhibitors ethanol and histidine, combined with the metal-ion chelator ethylenediaminetetraacetic acid (EDTA), were determined to be effective stabilizers of Ad5. Arrhenius plots of stability data are consistent with a first-order inactivation mechanism with apparent activation energies for virus inactivation of 26.5 +/- 0.9 and 28.7 +/- 0.6 kcal/mol in the absence and presence of free-radical oxidation inhibitors, respectively. Optimization of formulation pH, as well as the EDTA and ethanol concentrations, allowed for the identification of formulations that further enhanced long-term storage stability. For example, Ad5 in an optimized liquid formulation (A195) lost <0.1 logs of infectivity after 24 months of storage at 4 degrees C. The immunogenicity of a recombinant Ad5-based human immunodeficiency virus (HIV) vaccine candidate expressing HIV-1 gag (MRKAd5gag) formulated in A195, was shown to be equivalent to the same vaccine formulated in A105. Therefore, the use of EDTA, ethanol, and histidine did not significantly alter the immunogenicity of the vaccine in mice. The identification of 4 degrees C stable liquid formulations should significantly enhance the utility of Ad5 as a vector for vaccines and gene therapy. PMID- 15349957 TI - In vitro characterization of PEGylated phospholipid micelles for improved drug solubilization: effects of PEG chain length and PC incorporation. AB - Sterically stabilized micelles (SSM) composed of poly(ethylene glycol-2000) grafted distearoylphosphatidylethanolamine (DSPE-PEG) and sterically stabilized mixed micelles (SSMM) composed of DSPE-PEG and egg-phosphatidyl choline (PC) have recently been introduced as novel lipid based carriers for water-insoluble drugs. However, factors that affect the solubilization behavior of these phospholipid micelles are not well understood. This study investigates the effect of PEG chain length and PC content on physical properties and solubilization potential of PEGylated phospholipid micelles. Critical micelle concentrations (CMC) determined for DSPE-PEG with different PEG chain lengths (2000, 3000, and 5000) using a fluorescent probe were in the micromolar range (0.5-1.5 microM) with higher CMC for longer PEG chain length. The size of micelles determined by quasi-elastic light scattering (QELS) showed that micellar systems became heterogeneous when PC was added at > or =25% for DSPE-PEG 2000 and > or = 40% for DSPE-PEG 5000, respectively. Above these critical PC ratios a significant increase in aggregation number and formation of rodlike particles were observed by small angle neutron scattering (SANS). Solubilization of diazepam was greater with DSPE PEG 2000 than DSPE-PEG 5000 simple micelles as determined by RP-HPLC. However, DSPE-PEG 5000 micelles showed greater improvement in solubilization of the water insoluble drug with an increase in PC content. In conclusion, phospholipid micelle size and solubilization potential varied with PEG chain length and PC content in the mixed micelle. Aggregation number and shape of the micelles did not significantly change until the critical PC concentrations. PMID- 15349958 TI - Using real-time quantitative TaqMan RT-PCR to evaluate the role of dexamethasone in gene regulation of rat P-glycoproteins mdr1a/1b and cytochrome P450 3A1/2. AB - Cytochromes P450 (CYPs) and p-glycoproteins (Pgps) are believed to play important roles in drug absorption, metabolism, and elimination. Numerous drugs and environmental chemicals can modulate expression of these two classes of genes in different species. The present study investigated the effect of dexamethasone (Dex) on gene expression on both message and protein levels of mdr1a, mdr1b, CYP3A1, and CYP3A2 in small intestine, colon, liver, kidney, and brain microvessels of the rats treated orally with Dex at 1 or 20 mg/kg/day for 3 days. The basal expression of mdr1a mRNA was highest in the brain microvessels followed by colon, small intestine, liver, and kidney, and mdr1b mRNA was highest in the brain microvessels followed by kidney, liver, colon, and small intestine. After Dex treatment, mdr1a mRNA was increased by 5.5- and 10.7-fold in the small intestine, decreased extensively by 85-90% in the liver, and showed little or no change in the colon, kidney, and brain microvessels compared to the control rats. A similar pattern was observed for mdr1b mRNA. CYP3A1 mRNA was increased in all tissues examined. CYP3A2 mRNA was not significantly changed with the exception that at 20 mg/kg CYP3A2 mRNA was increased 5- and 30-fold in the colon and kidney. In general, Western blot analyses were consistent with mRNA changes. CYP3A protein expression was increased in all tissues examined. The disparity of the impact of Dex on the CYP 3A and Pgp expression in these studies suggest that the regulation of Pgp expression is very complex and is difficult to predict solely based on the PXR response to xenobiotics. PMID- 15349959 TI - Radiopharmaceutical development of a freeze-dried kit formulation for the preparation of [99mTc-EDDA-HYNIC-D-Phe1, Tyr3]-octreotide, a somatostatin analog for tumor diagnosis. AB - [(99m)Tc-EDDA-HYNIC-D-Phe(1),Tyr(3)]-Octreotide ((99m)Tc-EDDA/HYNIC-TOC) is a promising new radiopharmaceutical with the potential to replace [(111)In-DTPA-D Phe(1)]-Octreotide ((111)In-DTPA-OCT) as the radiopharmaceutical for somatostatin receptor scintigraphy due to the advantage of improved image quality, lower radiation dose for the patient, and daily availability. Here we describe the development of a freeze-dried kit formulation based on the Tricine/EDDA exchange labeling approach for the preparation of this radiopharmaceutical in a clinical setting. Three parameters were of major importance to achieve a suitable formulation with a radiochemical purity (RCP) >90%: addition of bulking agent, the pH of the freeze-drying solution, and the content of stannous chloride. The final formulation consisted of 20 mg Tricine, 10 mg EDDA, 50 mg Mannitol, 20 microg SnCl(2). 2H(2)O, and 20 microg [HYNIC-D-Phe(1), Tyr(3)]-Octreotide (HYNIC TOC). Radiolabeling was performed by addition of 0.2 M Na(2)HPO(4) to adjust the pH to 6-7, followed by 0.5-2 GBq (99m)Tc sodium pertechnetate, in a total volume of 2 mL and incubation for 10 min in a boiling water bath. Mean RCP values of 10 batches showed values >90% over a storage period of up to 1 year, a high stability up to 24 h of the final preparation, and retained biological activity. The developed kit formulation forms the basis for further clinical evaluation of this promising new radiopharmaceutical. PMID- 15349960 TI - Usefulness of a novel Caco-2 cell perfusion system. I. In vitro prediction of the absorption potential of passively diffused compounds. AB - A simple, reliable, and user friendly system was established to cultivate Caco-2 cell monolayer for epithelial transport studies. After an initial growth period of 1 week in a CO(2) incubator, Caco-2 cells were cultivated in an automated continuous perfusion system (Minucells and Minutissue, Germany). Medium was constantly renewed at the apical and basal side of the monolayers, which resulted in a continuous supply of nutrients as well as in a continuous removal of metabolite wastes. The monolayers obtained with the new perfusion culture system were evaluated to estimate the passive transport properties of a series of model compounds. The results produced were compared to those of monolayers obtained with the standard 21-day system. The integrity of cell monolayers was checked by measuring transepithelial electrical resistance (TEER) and by the transport of the paracellular leakage marker sodium fluorescein. The results of confocal microscopy as well as TEER measurements indicated the formation of a monolayer on various support filters. The growth and differentiation of Caco-2 cells were highly dependent upon the individual support filters and extracellular matrix proteins used for Caco-2 attachment. The permeability coefficients of several model compounds across Caco-2 cells obtained with the perfusion system were approximately two-fold higher than those obtained using the traditional 21-day Snapwell-based cultures. A good correlation was found between the transport of passively diffused drugs across Caco-2 monolayers differentiated in the perfusion system and the transport according to the standard method. The rank ordering of high permeable model compounds tested through Caco-2 monolayers, differentiated in a perfusion system, was similar to the standard 21-day culture method. PMID- 15349961 TI - A comprehensive analysis of the role of correction factors in the allometric predictivity of clearance from rat, dog, and monkey to humans. AB - This study was conducted to comprehensively evaluate the performance of various allometric scaling methods for the prediction of human clearance. Allometric scaling was used to predict clearance for 103 compounds, for which clearance data in the rat, dog, monkey, and humans were available. Allometry was performed using all three preclinical species and with combinations of any two species. The methods employed included standard allometry and various correction factors, including brain weight, maximum lifespan potential, and glomerular filtration. Scaling was performed on all compounds universally and on segregated subsets based on allometric exponent, clearance, physicochemical property, or route of elimination. 776 allometric combinations with 27,313 individual outcomes were performed. A predicted-to-observed clearance ratio of 0.5 to twofold was preselected as the criterion for predictive success. The success rate of allometric scaling ranged from 18 to 53%; none of the correction factors resulted in substantially improved predictivity. Furthermore, none of the methods attempted in this study achieved a success rate greater than that observed by simply estimating human clearance based on monkey hepatic extraction. Prospective allometric scaling, with or without correction factors, represents a suboptimal technique for estimating human clearance based on in vivo preclinical data. PMID- 15349962 TI - Quantitative structure-pharmacokinetic parameters relationships (QSPKR) analysis of antimicrobial agents in humans using simulated annealing k-nearest-neighbor and partial least-square analysis methods. AB - We have developed quantitative structure-pharmacokinetic parameters relationship (QSPKR) models using k-nearest-neighbor (k-NN) and partial least-square (PLS) methods to predict the volume of distribution at steady state (Vss) and clearance (CL) of 44 antimicrobial agents in humans. The performance of QSPKR was determined by the values of the internal leave-one-out, crossvalidated coefficient of determination q(2) for the training set and external predictive r(2) for the test set. The best simulated annealing (SA)-kNN model was highly predictive for Vss and provided q(2) and r(2) values of 0.93 and 0.80, respectively. For all compounds, the model produced average fold error values for Vss of 1.00 and for 93% of the compounds provided predictions that were within a twofold error of actual values. The best SA-kNN model for prediction of CL yielded q(2) and r(2) values of 0.77 and 0.94, respectively, and had an average fold rror of 1.05. Use of PLS methods resulted in inferior QSPKR models. The SA kNN QSPKR approach has utility in drug discovery and development in the identification of compounds that possess appropriate pharmacokinetic characteristics in humans, and will assist in the selection of a suitable starting dose for Phase I, first-time-in-man studies. PMID- 15349963 TI - Ciclopirox delivery into the human nail plate. AB - The human nail penetration of the antifungal ciclopirox was determined for marketed gel containing 0.77% of ciclopirox, an experimental gel containing 2% of ciclopirox, and a marketed lacquer containing 8% of ciclopirox. After 14 days dosing, unabsorbed drug remaining on the surface, drug within the infection-prone area, and the amount that had penetrated through the nail were determined. Ciclopirox delivery into and through the nail was significantly greater from the marketed gel, than from either the experimental gel or the nail lacquer (p < 0.05). In addition, the surface nail contained more unabsorbed drug from the lacquer. Further, the drug penetrating into and through the nail was also greater from the marketed gel, leading to a higher Calculated Efficacy Coefficient for the marketed gel, than from the marketed lacquer or the experimental gel. The formulation plays an important role in the enhancement of ciclopirox permeation into and through the human nail plate, and the concentration of ciclopirox in the formulation was not a factor in determining penetration. PMID- 15349964 TI - Effect of metal cations on the conformation and inactivation of recombinant human factor VIII. AB - Heavy metals have been implicated in the aggregation of proteins and the pathophysiology of several neurodegenerative diseases. Herein, we describe the interaction of recombinant human factor VIII (rhFVIII) with Al(+3), Tb(+3), Co(+2), and Fe(+3) using a combination of intrinsic fluorescence, circular dichroism, and high-resolution fourth-derivative absorbance analysis. rhFVIII in solution was titrated with the metal cations and the properties of the resulting complexes were examined. rhFVIII has a tendency to aggregate and inactivate slowly over time under physiological conditions, but this aggregation process is greatly accelerated in the presence of metals with Al(+3) being the most efficient. This leads to a complete loss of activity of the protein. Al(+3) induced conformational changes in the protein were small but detectable with limited changes seen in secondary and tertiary structure. Because rhFVIII is a multidomain protein with subunits linked through divalent metal cations, the small intramolecular changes seen may be attributed to rearrangements of the subunits to an aggregation-competent conformer that is very similar to that of the native form. PMID- 15349965 TI - Characterization of renal excretion mechanism for a novel diuretic, M17055, in rats. AB - M17055 was developed as a novel diuretic that inhibits both Na(+), K(+), and 2Cl( ) cotransport at the thick ascending Henle's loop and Na(+) reuptake at the distal tubule. It is secreted at the renal proximal tubules. The purpose of the present study was to characterize the renal excretion mechanism of M17055. We used the renal cortical slices and brush border membrane vesicles (BBMVs) to investigate the transport mechanisms across the basolateral and brush border membranes, respectively. M17055 uptake by rat renal slices increased with time and was saturable. Several organic anions including probenecid, para aminohippurate (PAH), and estrone-3-sulfate, decreased M17055 uptake. The uptake of M17055 was also observed into HEK293 cells expressing rat OAT1, and was inhibited by PAH. M17055 uptake by BBMVs was time-dependent, saturable, osmolarity-sensitive, and inhibited by several organic anions, but not by PAH. These results suggest that plural organic anion transport systems are involved in M17055 transport via both basolateral and brush border membranes of proximal tubule epithelial cells, a part of the renal uptake being mediated by OAT1. PMID- 15349966 TI - In vitro absorption and secretory quotients: practical criteria derived from a study of 331 compounds to assess for the impact of P-glycoprotein-mediated efflux on drug candidates. AB - The absorptive (AQ) and secretory (SQ) quotients have been proposed as a novel experimental approach to quantify the modulation of intestinal absorption and secretion by P-glycoprotein (Pgp). Because these unidirectional assays inherently assess for the impact of Pgp, conclusions as to whether a compound is a Pgp substrate will be made from the data. Therefore, the objective of this study was to establish the relationship between AQ/SQ and the bidirectional efflux assay and to derive criteria to classify a compound as a Pgp substrate. AQ and SQ parameters were calculated for 331 compounds that had previously been evaluated in the bidirectional assay and the concordance of Pgp substrate classification between these methods assessed by establishing AQ/SQ criteria of increasing magnitude. The AQ and SQ values correctly identified 80 and 85% of the compounds as Pgp substrates/nonsubstrates relative to the bidirectional efflux assay. This study demonstrates that the optimal AQ and SQ value to classify compounds as Pgp substrates was 0.3 and provides a basis to deploy unidirectional efflux assays in the early stages of drug discovery, which would benefit from the twofold increase in throughput over current bidirectional transport assays. PMID- 15349967 TI - Degradation kinetics of high molecular weight poly(L-lactide) microspheres and release mechanism of lipid:DNA complexes. AB - Plasmid DNA encoding the green lantern protein was ion-paired with 1,2-dioleoyl, 3-trimethylammonium propane (DOTAP) at a (+/-) charge ratio of (1:1) to form a hydrophobic ion-pair (HIP) complex using the Bligh and Dyer method, and transferred into methylene chloride. Precipitation with a compressed antisolvent (PCA) was then employed to encapsulate plasmid DNA into poly(L-lactide) (PLLA) microspheres. The hydrophobicity of DOTAP:DNA complexes allowed consistently high encapsulation efficiencies (>70%) to be achieved. Release of the DOTAP:DNA complex from PLLA microspheres exhibited minimal burst and a short (ca. 1 week) lag phase, followed by sustained release over a 20 week period. Release kinetics were consistent with a simple Fickian diffusion model. No correlation was identified between release rate of soluble poly(L-lactide) species (< or =10 lactate units) from PLLA and the DNA release kinetics. Only approximately 12% of the polymer was degraded into soluble poly(L-lactide) over the time frame where approximately 90% of the plasmid load had been released. PMID- 15349968 TI - In vitro and in vivo evaluation of a sulfobutyl ether beta-cyclodextrin enabled etomidate formulation. AB - In this study, we report the formulation and in vivo evaluation of etomidate in an aqueous solution using sulfobutyl ether-7 beta-cyclodextrin (SBE-CD, Captisol) as a solubilizing agent. The phase-solubility behavior of etomidate as a function of SBE-CD concentration was evaluated, and accelerated solution stability studies of 2 mg/mL etomidate in a 5% w/v SBE-CD solution were conducted. The intravenous administration of the SBE-CD etomidate formulation in dogs was compared with Amidate, the commercial etomidate drug product formulated with propylene glycol as a cosolvent. The etomidate plasma concentration-time data were fit to a three compartment mamillary model and the derived standard pharmacokinetic parameters were not statistically different between the two formulations (n = 4, p > 0.050). Concurrent pharmacodynamic analysis provided statistically equivalent maximum effects and median inhibitory concentrations for the two formulations. In vivo hemolysis after intravenous administration of Amidate was 10-fold higher than the SBE-CD formulation. Whereas Amidate cannot be given subcutaneously because of the cosolvent in the formulation, a 12 mg/mL aqueous solution of etomidate in 20% (w/v) SBE-CD was well tolerated by this route. The results suggest that the SBE CD formulation is a viable clinical drug product with a reduced side-effect profile. PMID- 15349969 TI - Avidin/biotin-liposome system injected in the pleural space for drug delivery to mediastinal lymph nodes. AB - The objective of this study was to develop a more effective liposome-based method for delivering drugs to mediastinal nodes. Nodal uptake was determined after intrapleural injection of the avidin/biotin-liposome system in normal rats. The effect of injection sequence (avidin injected 2 h before biotin-liposomes and vice versa), volume injected, and administered dose of the agents is described. Pharmacokinetics of the avidin/biotin-liposome system was monitored with scintigraphic imaging by labeling the biotin-liposomes with technetium-99m ((99m)Tc). To identify the nodes during the biodistribution studies, patent blue dye was encapsulated in the biotin-liposomes. Tissue biodistribution studies were performed 22 h after injection of the (99m)Tc-blue-biotin-liposomes. When avidin was injected before (99m)Tc-blue-biotin-liposomes, better mediastinal node targeting (15.7%; p < 0.05) was achieved than when biotin-liposomes were injected first (8.3%) or when only biotin-liposomes were injected (1.0%). Injection of a small dose of liposomes (0.5 mg phospholipid) and avidin (0.5 mg) resulted in the most favorable drug delivery to mediastinal nodes and other organs. Intrapleural injection of the avidin/biotin-liposome system could potentially be used for drug delivery to disease processes such as lung cancer, anthrax, and tuberculosis that invade mediastinal nodes and use them as centers of incubation and dissemination. PMID- 15349970 TI - Retrospective statistical analysis of lyophilized protein formulations of progenipoietin using PLS: determination of the critical parameters for long-term storage stability. AB - Although certain criteria have become recognized as being essential for a stable lyophilized formulation, the relative importance of different stability criteria has not been demonstrated quantitatively. This study uses multivariate statistical methods to determine the relative importance of certain formulation variables that affect long-term storage stability of a therapeutic protein. Using the projection to latent structures (PLS) method, a retrospective analysis was conducted of 18 formulations of progenipoietin (ProGP), a potential protein therapeutic agent. The relative importance of composition, pH, maintenance of protein structure (as determined by infrared (IR) spectroscopy), and thermochemical properties of the glassy state (as measured by differential scanning calorimetry (DSC)) were evaluated. Various stability endpoints were assessed and validated models constructed for each using the PLS method. Retention of parent protein and the appearance of degradation products could be adequately modeled using PLS. The models demonstrate the importance of retention of native structure in the solid state and controlling the pH. The relative importance of T(g) in affecting storage stability was low, as all of the samples had T(g) values above the highest storage temperature (40 degrees C). However, other indicators of molecular mobility in the solid state, such as change in DeltaC(p) upon annealing, appear to be important, even for storage below T(g). For the first time, the relative importance of certain properties in controlling long-term storage stability could be assessed quantitatively. In general, the most important parameters appear to be pH and retention of native structure in the solid state. However, for some stability endpoints, the composition (concentration of protein or various excipients), as well as some DSC parameters, were found to be significant in predicting long-term stability. PMID- 15349971 TI - Relationship between encapsulated drug particle size and initial release of recombinant human growth hormone from biodegradable microspheres. AB - Protein microencapsulation in biodegradable polymers is a promising route to provide for sustained release. One important characteristic in this regard is the size of the particles encapsulated within the microspheres. In this investigation, we have employed spray-freeze drying to generate particles for encapsulation, and examined the effect of various atomization conditions. Conditions were identified resulting in minimization of the particle size for the therapeutic protein recombinant human growth hormone (rhGH). The polymer employed was poly(lactide-co-glycolide) (PLG). The greatest friability for the powder, and hence smallest particle size (e.g., sub-micron), was achieved as the mass flow ratio of atomization (air to liquid) was increased. Protein powders over a range of particle sizes were encapsulated in biodegradable microspheres using a cryogenic, non-aqueous process. The initial release (both in vitro and in vivo) from these batches was found to decrease with decreasing encapsulated protein particle size; these findings are consistent with the percolation theory. Hence, judicious selection of process variables to reduce the particle size of rhGH is one strategy that can be used to minimize initial release of the microencapsulated protein. PMID- 15349972 TI - Mechanism of glidants: investigation of the effect of different colloidal silicon dioxide types on powder flow by atomic force and scanning electron microscopy. AB - The effect of hydrophilic and hydrophobic colloidal silicon dioxide types (CSD) on the flow characteristics of microcrystalline cellulose (MCC) under different mixing conditions was macroscopically measured using the angle of repose method, the bulk and tapped densities. CSD ameliorated the flow characteristics in general, but hydrophobic CSD was more effective compared to the hydrophilic types under gentle mixing conditions. The macroscopic effect was explained on the particle level by scanning electron (SEM) and atomic force microscopy (AFM) studies. The CSD distribution on the MCC surface was more uniform for the hydrophobic type and was independent from the mixing conditions used in this study. From the cumulative adhesion force distributions of the mixtures, determined by AFM, the mean and the standard deviation of the adhesion force were calculated. The means were 44.8 nN for MCC alone, 25.2 and 28.3 nN for mixtures containing the two hydrophilic types, and 13.8 N for the hydrophobic CSD under gentle mixing conditions in a Turbula mixer. Stronger mixing in a plowshare mixer led to a further reduction to 17.5 and 17.4 nN for the two hydrophilic types, while the hydrophobic CSD showing a value of 13.9 nN was unchanged. A linear correlation between the angle of repose and the adhesion force could be established, indicating that for routine measurements of the efficiency of a glidant the simple angle of repose method is sufficient. PMID- 15349973 TI - Immunohistochemical visualization of corticotropin-releasing factor type 1 (CRF1) receptors in monkey brain. AB - Corticotropin-releasing factor receptor type 1, CRF1, plays a prominent role in the hypothalamic-pituitary-adrenal (HPA) axis and is implicated in the autonomic and behavioral responses to stress. Dysregulation of the CRF system may underlie the pathophysiology of several disorders, including depression and anxiety. The distribution of CRF1 mRNA and CRF1 specific ligand binding has been reported by multiple groups in rodents using in situ hybridization and receptor autoradiography, respectively. More recently, somewhat conflicting rodent anti CRF1 immunohistochemical studies were reported. In this study we report the generation of an antihuman CRF1 antiserum and provide the first immunohistochemical description of CRF1 distribution in a primate brain, that of the rhesus monkey. The specificity of anti-CRF-R1 antiserum R221 was demonstrated using transfected hCRF1-expressing HEK 293 cells and rhesus monkey pituitary. CRF1-immunoreactive neurons were widespread in the rhesus brain. CRF1 staining was associated with neuronal cell bodies and dendrites and was primarily intracellular, suggesting a high rate of receptor turnover or receptor sequestration. Anti-CRF1 immunoreactivity was most abundant in pituitary, cerebellum, and in portions of brain stem associated with sensorimotor function. CRF1 staining was also observed in cerebral cortex, basal forebrain, portions of the basal ganglia, and thalamus. Staining was relatively low in prefrontal cortex and in limbic areas, which may reflect masking of the N-terminal epitope. The distribution of CRF1 immunoreactivity is suggestive of roles in attentional processing as well as the processing of motor and sensory information. PMID- 15349974 TI - Remodeling of a larval skeletal muscle motoneuron to drive the posterior cardiac pacemaker in the adult moth, Manduca sexta. AB - During postembryonic development, a larval skeletal muscle motoneuron, MN-1 in abdominal segments 7 and 8, becomes respecified to innervate the terminal cardiac chamber of adult Manduca sexta. Neural tracing techniques and electrophysiology were used in this study to describe the anatomical and physiological remodeling of this identified motoneuron. During metamorphosis the MN-1 in segments 7 and 8 undergoes dendritic reorganization. Long new dendrites extend anteriorly in the terminal ganglion neuropil. Intracellular and extracellular recordings showed that broader action potentials, increased firing rate, and development of a bursting activity pattern accompany MN-1 respecification. Cardiac mechanograms showed that MN-1 activity bursts always correlate with the anterograde cardiac beat. Bilateral MNs-1 fire at similar times to activate and sustain the putative cardiac pacemaker activity of the terminal chamber synergistically. After remodeling, MN-1 output could be influenced rapidly by sensory inputs during evoked cardiac reversal. The effect is exerted by inhibition of MN-1 firing that, in turn, causes early blockade of the anterograde beat and reversal to the retrograde direction of beat. PMID- 15349975 TI - Expression of Hsp27 in retinal ganglion cells of the rat during postnatal development. AB - The small heat shock protein Hsp27 has been shown to protect neurons from apoptosis. We have recently shown the expression of Hsp27 in a subset of injured adult retinal ganglion cells (RGCs), a response that is muted by the administration of brain-derived neurotrophic factor. This work has suggested a role for Hsp27 in the long-term survival of RGCs following injury. The purpose of this study was to investigate the expression of Hsp27 during postnatal retinal development, based on Hsp27's role as a neuronal survival factor and on its up regulation in the adult injured retina. Expression of Hsp27 in the developing retina was examined at various times postnatally (between P0 and P24) by using immunohistochemical techniques. We report that Hsp27 expression peaks in the ganglion cell layer between P6 and P12 and is not detected at earlier (P0-P3) or later (P15-P24) times. Double labeling of the Hsp27-positive cells with Fluorogold applied to the superior colliculus confirmed that Hsp27-positive cells in the ganglion cell layer are RGCs. We have shown developmentally regulated expression of Hsp27 in RGCs of the postnatal rat. The retinal expression of Hsp27 correlates temporally with innervation of the tectum by late-born RGCs and with onset of spontaneous retinotectal activity. We propose that the expression of Hsp27 may play an important role in retinal development during a critical period of RGC functional connectivity with the superior colliculus. PMID- 15349977 TI - Projection patterns of posterior dorsal unpaired median neurons of the locust subesophageal ganglion. AB - Six neurons in a group of dorsal unpaired median (DUM) neurons with cell bodies in the posterior part-maxillary and labial neuromeres-of the subesophageal ganglion of locusts have two axons each that descend into both the left and the right halves of the ganglia of the ventral nerve cord. None of the neurons has peripheral axons, so they are interneurons. Electrophysiology shows that the axons of at least four neurons project to the terminal abdominal ganglion to which they conduct spikes at a velocity of 0.5-0.6 m. second(-1). In the somata, the spikes have a smaller amplitude and briefer duration at half height than the spikes of thoracic, efferent DUM neurons. Each neuron has bilaterally symmetrical branches within the subesophageal ganglion and in the thoracic ganglia. On the basis of the specific patterns of branches, and the neuropiles, tracts, and commissures in which they occur, three types of neurons (DUM SD 1-3) can be recognized. DUM SD 1 and 3 project to ventral regions of neuropile in the thoracic ganglia in which the efferent DUM neurons of these ganglia have no branches. DUM SD 2 projects to dorsal neuropiles. The projection patterns of these putatively octopaminergic neurons suggest that they could be the source of the octopaminergic modulation of networks underlying sensory processing and motor pattern generation within these ganglia. Within this group of posterior DUM neurons, two additional cells were stained that have axons ascending to the brain. PMID- 15349976 TI - Retinal abnormalities associated with the G90D mutation in opsin. AB - Several mutations in the opsin gene have been associated with congenital stationary night blindness, considered to be a relatively nonprogressive disorder. In the present study, we examined the structural and functional changes induced by one of these mutations, i.e., substitution of aspartic acid for glycine at position 90 (G90D). Transgenic mice were created in which the ratio of transgenic opsin transcript to endogenous was 0.5:1, 1.7:1, or 2.5:1 and were studied via light and electron microscopy, immunocytochemistry, electroretinography (ERG), and spectrophotometry. Retinas with transgenic opsin levels equivalent to one endogenous allele (G0.5) appeared normal for a period of about 3-4 months, but at later ages there were disorganized, shortened rod outer segments (ROS), and a loss of photoreceptor nuclei. Higher levels of G90D opsin expression produced earlier signs of retinal degeneration and more severe disruption of photoreceptor morphology. Despite these adverse effects, the mutation had a positive effect on the retinas of rhodopsin knockout (R-/-) mice, whose visual cells fail to form ROS and rapidly degenerate. Incorporation of the transgene in the null background (G+/-/R-/- or G+/+/R-/-) led to the development of ROS containing G90D opsin and prolonged survival of photoreceptors. Absorbance spectra measured both in vitro and in situ showed a significant reduction of more than 90% in the amount of light-sensitive pigment in the retinas of G+/+/R-/- mice, and ERG recordings revealed a >1 log unit loss in sensitivity. However, the histological appearances of the retinas of these mice show no significant loss of photoreceptors and little change in the lengths of their outer segments. These findings suggest that much of the ERG sensitivity loss derives from the reduced quantal absorption that results from a failure of G90D opsin to bind to its chromophore and form a normal complement of light-sensitive visual pigment. PMID- 15349978 TI - Regulation of GABA transporter mRNA and protein after photothrombotic infarct in rat brain. AB - Animal models of focal ischemic infarcts reveal an impaired GABAergic (gamma aminobutyric acid) neurotransmission. GABA, the main inhibitory neurotransmitter, is primarily taken up by specific sodium-dependent transporters. As these transporters play a crucial role in maintaining levels of GABA concentration, they may be functionally involved in ischemic processes. We investigated whether the mRNA and protein expression of GAT-1, the dominant neuronal GABA transporter, is altered after cortical infarct induced by photothrombosis in Wistar rats. In situ hybridization was performed to analyze GAT-1 mRNA-positive cells in cortical brain regions and the hippocampus. The lesion dramatically raised the number of GABA transporter mRNA-expressing cells in all investigated cortical regions. Double-labeling studies with a general neuronal marker and a marker for astrocytes revealed that cells expressing GAT-1 mRNA after photothrombosis are neurons. The mRNA expression pattern of all hippocampal subfields remained unchanged. In contrast, cortical GAT-1 protein density was only slightly affected and surprisingly in the opposite way. In the primary and secondary somatosensory cortex, density values were significantly reduced. Immunoreactivity was not altered in all investigated hippocampal areas. We found a marked discordance between the increased number of cells expressing GAT-1 mRNA in the cortex and the reduced tissue GAT-1 protein content. Focal brain ischemia obviously triggers mechanisms that interfere with GAT-1 transcriptional regulation and protein synthesis or turnover. PMID- 15349979 TI - Distribution and development of glutamic acid decarboxylase immunoreactivity in the spinal cord of the dogfish Scyliorhinus canicula (elasmobranchs). AB - The adult distribution and development of gamma-aminobutyric acid (GABA) synthesizing cells and fibers in the spinal cord of the lesser spotted dogfish (Scyliorhinus canicula L.) was studied by means of immunohistochemistry using antibodies against glutamic acid decarboxylase (GAD). Complementary immunostaining with antibodies against GABA, tyrosine hydroxylase (TH), and HuC/HuD (members of the Hu/Elav family of RNA-associated proteins) and staining with a reduced silver procedure ("en bloc" Bielschowski method), Nissl, and hematoxylin were also used. In adults, GAD-immunoreactive (GAD-ir) cells were observed in the ventral horns, in the spinal nucleus of the dorsal horn, at the base of the dorsal horns, and around the central canal, where some GAD-ir cells were cerebrospinal fluid-contacting (CSF-c). In addition, a few GAD-ir cells were observed in the lateral funiculus between the ventral horn and the marginal nucleus. The adult spinal cord was richly innervated by GAD-ir fibers. Large numbers of GAD-ir fibers and boutons were observed in the dorsal and ventral horns and also interstitially in the dorsal, lateral, and ventral funiculi. In addition, a rich GAD-ir innervation was observed in the marginal nucleus of the spinal cord. In the embryonic spinal cord, GAD-ir cells develop very early: The earliest cells were observed in the very thin mantle/marginal layer of stage 22 embryos in a short length of the spinal cord. At stages 25 and 26, several types of GAD-ir cells (commissural and noncommissural) were distinguished, and two of these cells were of CSF-c type. At stages 28, 30, and 31, the GAD-ir populations exhibited a marked longitudinal columnar organization. Double-immunolabeling experiments in embryos showed the presence of two different GAD-ir CSF-c cell populations, one ventral that is simultaneously TH-ir and other more dorsal that is TH-negative. By stage 33 (prehatching), GAD-expressing cells are present in virtually all loci, as in adults, especially in the ventral horn and base of the dorsal horn. The present results for the lesser spotted dogfish suggest an important role for gamma-aminobutyric acid in sensory and motor circuits of the spinal cord. PMID- 15349980 TI - Evaluation of NMP179 for the detection of squamous intraepithelial neoplasia in ThinPrep cervical slides using a combination of double immunostaining and morphometric methods of analysis. AB - This study investigates the potential value of the nuclear matrix protein NMP179 as a marker of abnormal squamous cells in ThinPrep slides. Forty-six cervical scrapes were collected as cell suspensions and ThinPrep slides were prepared. They were double-immunostained for NMP179 and Cytokeratin 18 (CK18), an endocervical cell marker. The method of analysis adopted for the study was designed to distinguish the abnormal squamous cells from benign epithelial cell so that the percentages of abnormal squamous cells that expressed the marker could accurately be determined. Initially, an attempt was made to identify benign and abnormal cells in the ThinPrep slides on the basis of their morphology and immunostaining patterns. Discrimination between the various types of epithelial cells was incomplete using this approach and a more precise method of discrimination between the different epithelial cell types was carried out using a combination of double immunostaining (NMP179 and CK18) and morphometry using nuclear area and nuclear cytoplasmic ratios. Once the different epithelial cell types had been identified, the specificity and sensitivity of NMP179 were determined. The optimal sensitivity (89.9%) was achieved at the N/C ratio 0.36; however, the specificity of NMP179 was very low for all N/C ratios and ranged from 38.8% to 42.2%. PMID- 15349981 TI - Rapid review of liquid-based smears as a quality control measure. AB - The objective of this study was to investigate the effectiveness of a standardized method of rapid review (RR) of monolayer preparations for the identification of abnormalities, the presence of an endocervical component and infectious agents. A total of 200 ThinPrep (Cytyc, Boxborough, MA) slides representing the spectrum of abnormalities commonly encountered in cervical/vaginal cytologic specimens was retrieved from archive. The study set comprised 129 cases within normal limits (WNL); 36 low-grade epithelial abnormalities (LGEA); 28 high-grade epithelial abnormalities (HGEA), including 2 endocervical adenocarcinomas in situ (AIS) and 7 carcinomas. Eighteen false negative (FN) cases were also included for study. Originally missed on initial review, these cases were found to be abnormal on quality control review (17 LGEA; 1 AIS). Commonly encountered infectious agents were represented and included Candida albicans, Trichomonas vaginalis, herpes simplex virus, and Actinomyces. The slides were reviewed using a standardized method of RR (turret technique, for 60 sec) by three experienced screeners masked to the original reference diagnosis. Median sensitivity for LGEA was 70% (range, 67-72%); HGEA, 69% (range, 54-80%); and FN, 65% (range, 56-78%). Specificity remained high, median specificity for LGEA was 95%; HGEA, 97%; and FN, 100%. There was no significant overcalling of any diagnostic category. The chi-square test at P < 0.05 showed no significant difference between RR and full manual rescreen of the ThinPrep smears in this study. While no statistical difference was proven, the sensitivity measurements for all categories of abnormality were moderate due to the high proportion of atypical cases included into the study set. Abnormalities on the monolayer preparations frequently displayed fewer, smaller groups of disaggregated cells with rounded cytoplasmic outlines that were difficult to discern on RR. Interobserver variation was noted. Monolayers with a paucity of diagnostic cells and those displaying subtle nuclear atypia were often overlooked. PMID- 15349982 TI - Fine-needle aspiration of adult small-round-cell tumors studied with flow cytometry. AB - Immunophenotypic study is critical for the diagnosis of adult small-round-cell tumors (SRCTs). We describe three patients with Ewing's sarcoma/primitive neuroectodermal tumor (ES/PNET) and one patient with neuroblastoma in which flow cytometry immunophenotyping (FCI) on the fine-needle aspirate (FNA) and bone marrow aspirate (BMA) demonstrated an abnormal population of cells that were CD45(-) and CD16/CD56(+). Four patients with mean age of 30 years, three male and one female, clinically suspicious for a lymphoma or SRCT are described. FNA, BMA, and biopsy specimens were obtained for routine cytologic and histologic evaluation. Fresh tissue was studied by FCI. In all cases, the cytology smears showed small cells with round nuclei, slightly irregular nuclear membranes, fine chromatin, and scant cytoplasm. FCI showed CD16/56(+) and CD45(-) neoplastic cells in all cases. In one case, 76% of these cells were CD99(+). The diagnoses of ES/PNET were confirmed by immunohistochemical, ultrastructural, and cytogenetic studies. ES/PNET in FNA and BMA can be efficiently and rapidly diagnosed by combining cytologic examination with FCI using a panel including CD45, CD16/56, and CD99. PMID- 15349983 TI - Influence of specimen adequacy on the diagnosis of ASCUS. AB - The impact of specimen adequacy on the diagnosis of cervical cytology is not fully understood. Virtually, the greatest impact of smear quality limitation factors is on the diagnosis of atypical squamous cells of undetermined significance (ASCUS) because this category is more susceptible to interpretation mistake due to its criteria subjectivity. This study aims to evaluate the impact of smear quality impairing factors on the diagnosis of ASCUS. The 1,507 cases of ASCUS studied were obtained from the Cervical Cancer Screening Program of Parana (CCSPP). The cases were reviewed by expert cytopathologists and the results were correlated to the specimen adequacy. The results showed that the presence of obscuring blood impairs the correct diagnosis and leads the pathologist to overdiagnose negative smears as ASCUS. The presence of adequate endocervical sampling contributes to a higher rate of correct ASCUS diagnoses but did not influence the pathologist to over- or underestimate the cytological findings. PMID- 15349984 TI - The role of flow cytometric immunophenotyping in improving the diagnostic accuracy in referred fine-needle aspiration specimens. AB - Flow cytometric (FCM) immunophenotyping has an important role in the diagnostic work up of fine-needle aspiration (FNA) specimens obtained from lymphoid lesions. The objective of the present study was to evaluate the feasibility of this method with respect to referred FNA specimens. One hundred and two FNA specimens referred to our laboratory for FCM analysis during the last 3 years were studied. Specimens were sent, accompanied by cytological smears, from 11 distant hospitals by ordinary mail. The evaluation of potential B-cell monoclonality, the main diagnostic issue to be resolved using FCM, was possible in 86 of these 102 cases. The remaining 16 samples could not be analyzed or adequately interpreted because of sparse or necrotic material. A monoclonal B-cell population was found in 17/86 satisfactory cases, of which 16 were histologically confirmed. Eight cases contained cells positive for the epithelial marker Ber-EP4 and were diagnosed accordingly as carcinomas. FCM analysis of specimens obtained with a clinical question of Hodgkin lymphoma or T-cell lymphomas did not yield definitive data. The time lapse between sampling and analysis (12-84 hr) did not affect the results. This probably was due to the fact that all aspirates were taken in Roswell Park Memorial Institute (RPMI) cell medium, supplemented with 50% fetal calf serum. In conclusion, this retrospective study establishes that it is possible, in the majority of cases, to refer FNA material for FCM immunophenotyping by mail, and that results regarding B-cell clonality in the case of small-cell lymphomas are reliable also after a transportation period of 3 4 days. Carcinoma may be similarly diagnosed and a diagnosis of lymphoma may be excluded in reactive proliferations. Cases with only a few atypical cells or specimens from patients suspected of having Hodgkin lymphoma or T-cell lymphomas are not suitable for analysis by FCM. PMID- 15349985 TI - Solid and papillary epithelial neoplasm of the pancreas (SPENP): observation of "Chinese character" morphology on fine-needle aspiration (FNA). PMID- 15349986 TI - Cytologic three-dimensional imaging for the interpretation of staining profiles: application of confocal laser scanning microscopy. PMID- 15349987 TI - Pap test in a high-risk population comparison of conventional and liquid-base cytology. AB - Liquid-based cytology (LBC) is believed to have better sensitivity than conventional smears (CSs) and offers the possibility to perform molecular assay. The goal of this work was to study the performance of CS and LBC in a high-risk population and to compare the results with the hybrid capture (HC) II for high risk human papillomavirus (HPV). Samples were collected from selected women with clinical suspicions of low genital tract lesion at Perola Biygnton Hospital (Sao Paulo, Brazil). After CS preparation, the brush was introduced in the endocervix and a new sample was collected and rinsed in the preservative medium of the system. The residual material was used to HC2. From 925 cases, LBC was unsatisfactory in 4 (1.51%) cases and CS was unsatisfactory in 100 cases (10.81%); among theses cases HC2(+) reactions was observed in 54 (54%) CSs and 3 (21.4%) LBCs. Considering cases with atypia of undetermined significance (squamous and glandular), 85 (77.27%) cases from LBC and 44 (86.4%) from CS were positive for HC2 assay for high-risk DNA-HPV. The difference among the methods was not significant (P < 0.38). The diagnosis improvement of LBC in comparison with CS was 86% in satisfactory samples, 92.76% in undetermined atypical lesions (including glandular), 83% in positive low-grade squamous intraepithelial lesions (LSIL(+)), and 86.84% in high-grade SIL(+) (HSIL(+)). HC2(+) reactions were observed in 144 CS cases and 266 LBC cases with abnormalities. Our results have showed that LBC was superior to CS in a high-risk population to detect lesions with high concordance with HC2(+) reactions; CSs also exhibit a high concordance with HC2 assay but with inferior performance to detect lesions. PMID- 15349988 TI - Liesegang rings in hemorrhagic urine. AB - Myriad Liesegang rings were encountered in the urine of a 43-year-old patient who was diagnosed as having idiopathic hematuria, emanating from the left ureter or kidney. Erythrocytes and the Liesegang rings spontaneously disappeared from the urine within 4 weeks. We are unaware of a report of a similar observation. Lack of descriptions may be due to underreporting or reflect true rarity of Liesegang rings in the urine. It is suggested that Liesegang rings may precipitate in the hemorrhagic urine as they do in diverse blood-imbibed tissues. PMID- 15349989 TI - Fine-needle aspiration cytology findings from a case of pancreatic heterotopia at the gastroesophageal junction. AB - Pancreatic heterotopia is a well-recognized entity occurring at a variety of sites in the gastrointestinal tract. However, only a handful of patients have been described with this abnormality at the gastroesophageal junction. Cytologic descriptions of pancreatic heterotopia in general are scant in the literature. We report the cytologic findings of ectopic pancreatic tissue at the gastroesophageal junction in a 41-year-old female, which was initially identified by endoscopic ultrasound and fine-needle aspiration. Cytologic examination of Papanicolaou-stained smears revealed clusters of benign-appearing ducts and small acini mixed with inflammatory cells. Hematoxylin and eosin staining of the Tru Cut biopsy revealed benign pancreatic tissue. The patient underwent surgical resection due to symptoms related to inflammation of the heterotopic pancreatic tissue. The increasing use of endoscopic ultrasound and fine-needle aspiration for the diagnosis of gastrointestinal tumors makes the recognition of pseudoneoplastic lesions at unusual sites an important step in optimum patient management. PMID- 15349990 TI - Nevus-cell aggregates in lymph nodes: fine-needle aspiration cytologic findings and resulting diagnostic difficulties. AB - We report a case of nodal nevus present in enlarged lymph nodes with changes of dermatopathic lymphadenopathy sampled by fine-needle aspiration (FNA) cytology prior to clinical evaluation of the patient. This lymph node pathology was established later by lymph node excisional biopsy, by which along with a skin biopsy the dermatopathic lymphadenopathy was tentatively attributed to early mycosis fungoides. The FNA revealed fairly atypical melanotic tissue from the dermatopathic lymphadenopathy along with nodules of uniform melanocytic nevoid cells, the presence of which in combination with the dermatopathic atypical tissue provided a tentative diagnosis of metastatic melanoma of unknown primary, with the diagnosis of nodal nevus presented as a less likely possibility. This is to our knowledge the first cytologic report on FNA of nodal nevus, which besides presenting cytologic findings of this entity highlights some of the problems related to providing an accurate diagnosis, if this exceptionally unusual pathologic entity is encountered in lymph nodes sampled for enlargement from pathologies unrelated to this entity. The subject of nevus changes in lymph nodes is briefly discussed. PMID- 15349992 TI - Salivary duct carcinoma with neuroendocrine features: report of a case with cytological and immunohistochemical study. AB - We report a salivary duct carcinoma (SDC) of parotid gland in a 75-year-old male. Initially, it was studied by fine-needle aspiration, which disclosed features of malignancy consistent with a high-grade carcinoma. Histologically, the tumor showed typical features of SDC, predominantly with a solid and apocrine pattern. The aggressive behavior of this tumor was documented by facial palsy and the presence of 12 regional lymph node metastases. Immunohistochemical study showed positivity for cytokeratins (AE1/AE3), cytokeratin 7, GCDFP-15, C-erbB-2, Mib-1, topoisomerase II alpha, p53, and androgen receptors. Diffuse positivity with chromogranin-A, synaptophysin, and Grimelius stains was also observed, suggesting endocrine features. Phosphotungstic acid hematoxylin, antimitochondrial antigen, progesterone and estrogen receptors, cytokeratin 20, and S-100 stains were negative. To our knowledge, this is the first case reported of SDC exhibiting neuroendocrine differentiation. PMID- 15349991 TI - Cytologic features of recurrent lymphoma involving the urinary bladder. AB - Recurrent lymphoma of the bladder only occasionally presents with genitourinary symptoms, and there are very few cases in the literature reporting the cytologic findings of involvement of the urinary bladder by lymphoma. We report the findings from a case of diffuse large B-cell lymphoma with immunoblastic morphology that was identified in a bladder barbotage specimen of a 77-year-old man who presented with recurrent urinary tract infection and hematuria. We describe the cytomorphological features of lymphoma cells in the urine and discuss the differential diagnoses. Correlation of cytologic findings with immunohistochemical results is crucial in the diagnosis of lymphoma involving the urinary bladder. PMID- 15349993 TI - Cytologic diagnosis of plasma myeloma in an 8-year-old child: case report. PMID- 15349994 TI - Correlation of immunocytochemistry and hybrid capture for human papillomavirus detection in cervical smears. PMID- 15349995 TI - Fine-needle aspiration cytology of Rosai-Dorfman disease. PMID- 15349996 TI - Revisiting liver transplant immunology: from the concept of immune engagement to the dualistic pathway paradigm. AB - Ever since the demonstration that allografts are rejected through immune reactions of the host, clinical therapies for organ allografts have relied on immune suppression to prevent these destructive events. A growing body of clinical and experimental data suggests that allografts elicit multiple, interactive immune responses. The result is not inevitably graft rejection, and "spontaneous" acceptance of fully allogeneic liver grafts occurs in rodents without immunosuppression. A spectrum of results range from spontaneous acceptance without immunosuppression to rejection with immunosuppression. The "dualistic pathway paradigm" aims to reconcile apparently conflicting observations in liver transplantation and proposes that: (1) immune engagement between the host and the allograft is instrumental in both rejection and acceptance; (2) there exist in all mammalian species congruent interactive pathways of immune activation whereby the fate of the allograft is determined by the quantitative results of these interactions; (3) the dualistic effect of immunosuppressive drugs on pathways of immune activation, conferring the capacity for favorable or unfavorable graft outcome should be investigated in experimental models in which organ allografts are spontaneously accepted. In conclusion the design of clinical strategies based on this research may contribute to protocols resulting in allograft acceptance without chronic immunosuppression. PMID- 15349997 TI - Experience after the evaluation of 700 potential donors for living donor liver transplantation in a single center. AB - Adequate selection of donors is a major prerequisite for living donor liver transplantation (LDLT). Few centers report on the entire number of potential donors considered or rejected for living donation. From April 1998 to July 2003, a total of 111 living donor liver transplantations were performed at our institution, with 622 potential donors for 297 adult recipients and 78 potential donors for 52 pediatric recipients evaluated. In the adult group, only 89 (14%) potential donors were considered suitable, with a total of 533 (86%) potential donors rejected. Of these, 67% were excluded either at initial screening or during the first and second steps of the evaluation procedure. In 31% of all cases, the evaluation of donors was canceled because of recipient issues. In the pediatric group, 22 (28%) donors were selected, with the other 56 (72%) rejected. Costs of the complete evaluation process accounted for 4,589 Euro (Euro) per donor. The evaluation of a potential living donor is a complex and expensive process. We present the results on the evaluation of the largest group of potential donors for adults reported in the literature. Only 14% of potential donors in our series were considered suitable candidates. It has not yet been established who should cover the expenses of the evaluation of all rejected donors. In conclusion, all efforts should be made in order to develop an effective screening protocol for the evaluation of donors with the aim of saving time and resources for a liver transplantation program. PMID- 15349998 TI - Evaluating the potential living liver donor. PMID- 15349999 TI - Molecular adsorbent recirculating system for acute and acute-on-chronic liver failure: a meta-analysis. AB - Molecular adsorbent recirculating system (MARS) is an important option for patients with liver failure to give them additional time for recovery or to serve as a "bridge" to transplantation. However, its effect on survival for such patients is not well known. Our aim was to assess the treatment effects of MARS on patients with acute and acute-on-chronic liver failure. The outcomes measure evaluated was survival. We searched Medline (1966-2002) and EMBASE (1974-2002) using the terms liver failure, liver support systems, and MARS. Our search was extended to the Cochrane Controlled Trials Registry Database, published abstracts from 5 international conferences, Teraklin (the manufacturer of MARS), known contacts, and bibliographies from each full-published report. We included trials published in English and non-English languages. Eligible studies were randomized and nonrandomized controlled trials, which compared the treatment effects of MARS with standard medical treatment. Of the 206 articles screened, 4 randomized controlled trials including 67 patients were analyzed. Two nonrandomized trials with 61 patients were used for explorative analysis. The methodology, population, intervention, and outcomes of each selected trial were evaluated by duplicate independent review. Disagreements were resolved by consensus. In the primary meta analysis, MARS treatment did not appear to reduce mortality significantly compared with standard medical treatment [relative risk (RR), 0.56; 95% confidence interval (CI), 0.28-1.14; P = .11]. Only 1 of the 4 randomized trials analyzed showed significant reduction in mortality. Sensitivity analysis of 3 peer-reviewed trials did not reduce mortality significantly with MARS treatment (RR, 0.72; 95% CI, 0.37-1.40; P = .33). Subgroup analysis of 2 trials for acute liver failure and another 2 trails for acute-on-chronic liver failure also did not reveal any benefit to survival with MARS treatment. In contrast, explorative analysis of 2 nonrandomized trials showed a significant survival benefit with MARS treatment (RR, 0.36; 95% CI, 0.17-0.76; P = .007). This was possibly related to bias in the selection of patients in the nonrandomized trials. In conclusion, MARS treatment had no significant survival benefit on patients with liver failure when compared with standard medical therapy. However, we found only a few trials with a small number of patients for the analysis, allowing for the possibility of false negative and erroneous conclusions. Well-conducted randomized trials are strongly recommended to define the role of MARS in the treatment of patients with liver failure. PMID- 15350000 TI - Meta-analysis in albumin dialysis: are we really ready for it? PMID- 15350001 TI - Pathophysiological effects of albumin dialysis in acute-on-chronic liver failure: a randomized controlled study. AB - The pathophysiological basis of acute-on-chronic liver failure (ACLF) is unclear but systemic inflammatory response is thought to be important. In patients with ACLF, the molecular adsorbents recirculating system (MARS) improves individual organ function, but the effect of MARS on the proposed mediators of systemic inflammatory response is unclear. The present study was designed to determine the effect of MARS on the cytokine profile, oxidative stress, nitric oxide, and ammonia. A total of 18 patients with alcohol-related ACLF due to inflammation related precipitants were randomized to receive standard medical therapy (SMT) alone, or with MARS therapy over 7 days. Plasma cytokines, malondialdehyde (MDA), free radical production, nitrate / nitrite (NOx), and ammonia were measured. Encephalopathy improved significantly with MARS (P < .01), but not with SMT. Mean arterial pressure and renal function remained unchanged. No significant change of plasma cytokines and ammonia levels were observed in either group. Plasma MDA levels did not change either. There was a fall in NOx (P < .05) with MARS, but not with SMT. In conclusion, in inflammation-related ACLF patients, albumin dialysis using MARS results in improvement of encephalopathy, independent of changes of ammonia or cytokines, without improving blood pressure or renal function. These results should temper the liberal use of MARS until further data is available. PMID- 15350002 TI - Long-term results and modeling to predict outcomes in recipients with HCV infection: results of the NIDDK liver transplantation database. AB - Hepatitis C virus (HCV)-associated liver disease is the most common indication for liver transplantation (LT). There are, however, no long-term (>5 year) studies of comparative outcomes for recipients with HCV infection, and no models capable of identifying recipients with HCV infection at greatest risk for adverse outcomes. We prospectively determined: 1) long-term outcomes, and 2) whether pretransplant patient or donor variables can be used to predict death and/or graft loss in recipients with HCV infection. A total of 165 HCV-infected recipients were eligible for this study. Pretransplant donor and recipient characteristics and patient and graft survival data were prospectively collected. Model building for outcomes was carried out using logistic regression. Receiver operating characteristic curves for different models were created and compared. Median follow-up was 8.5 years. Adjusted 10 year graft survival was 64% for recipients with HCV infection and 51% for uninfected recipients. A model incorporating pretransplant HCV ribonucleic acid (RNA), cytomegalovirus immunoglobulin (CMV IgG) serostatus, creatinine, bilirubin, prothrombin time international ratio (INR), recipient age, and donor age was developed to identify recipients at greatest risk of short-term mortality or graft loss (area under receiver operating characteristic curve = .83) In conclusion, long-term outcomes following LT for recipients with HCV infection are comparable to those for recipients undergoing LT for other indications. HCV-infected recipients at greatest risk for short-term mortality and graft loss can be identified using several readily identifiable pretransplant variables. Long-term death and graft loss specifically secondary to recurrence of HCV appears, however, to be determined primarily by factors other than those included in this analysis. PMID- 15350003 TI - Evolution of hepatitis C virus quasispecies immediately following liver transplantation. AB - Liver cirrhosis caused by chronic hepatitis C virus (HCV) infection is the main indication for liver transplantation (LT). There is little information on HCV genetic evolution following transplantation. The aim of this study was to carefully assess early evolution of HCV quasispecies in a cohort of 18 liver transplant recipients followed prospectively. Quasispecies analysis was performed by sequence analysis of the hypervariable region 1 (HVR1) before transplantation and at day 4 and week 4 following LT. A predominant variant was present in 12 (67%) of the 18 patients before transplantation and the same variant was propagated and remained predominant after LT in 6 (50%) of these patients. In the remaining individuals, there were major changes in the quasispecies composition, mostly occurring during the first days after LT. There was a progressive decrease in the nonsynonymous (dN)/synonymous (dS) ratios from baseline (1.2) to day 4 (.6) (P = .08) and to week 4 after LT (.3) (P = .015). Similarly, genetic distance (GD) declined from baseline (.1) to day 4 (.03) (P = .07) and to week 4 (.04) (P = .04). We did not find any differences in HCV genetic evolution between patients with mild (n = 10) or severe (n = 8) disease recurrence. In conclusion, during the first days following transplantation, HCV quasispecies becomes more homogenous, even after major changes in its composition. Importantly, these changes persist and even increase during the 1st month after transplantation. The "bottleneck" effect caused by the implantation of a new graft and the lack of selective pressure due to the strong immunosuppression most likely explain this particular pattern of genetic evolution. PMID- 15350004 TI - Herpes zoster after liver transplantation: incidence, risk factors, and complications. AB - Herpes zoster is the consequence of the reactivation of latent varicella-zoster infection. Immunosuppression may be a predisposing factor for herpes zoster. We have retrospectively assessed the risk of herpes zoster, the risk factors for its occurrence, and its evolution in a population of 209 consecutive liver transplant recipients. Herpes zoster developed in 25 (12%) of patients. One-, 3-, 5-, and 10 year actuarial rates of herpes zoster were 3%, 10%, 14%, and 18%, respectively. In a case-control study, patients developing herpes zoster were younger, received a higher number of immunosuppressive drugs, and were more frequently receiving mycophenolate mofetil or azathioprine. In multivariate analysis, the only factor related to herpes zoster occurrence was treatment with mycophenolate mofetil or azathioprine. Eight patients (31%) developed postherpetic neuralgia. In conclusion, herpes zoster is a relatively common complication after liver transplantation. It is related to immunosuppressive therapy. Postherpetic neuralgia develops in one third of patients with posttransplant herpes zoster. PMID- 15350005 TI - Increased prothrombin time and platelet counts in living donor right hepatectomy: implications for epidural anesthesia. AB - The risks and benefits of adult-to-adult living donor liver transplantation need to be carefully evaluated. Anesthetic management includes postoperative epidural pain relief; however, even patients with a normal preoperative coagulation profile may suffer transient postoperative coagulation derangement. This study explores the possible causes of postoperative coagulation derangement after donor hepatectomy and the possible implications on epidural analgesia. Thirty donors, American Society of Anesthesiology I, with no history of liver disease were considered suitable for the study. A thoracic epidural catheter was inserted before induction and removed when laboratory values were as follows: prothrombin time (PT) > 60%, activated partial thromboplastin time < 1.24 (sec), and platelet count > 100,000 mmf pound sterling (mm3). Standard blood tests were evaluated before surgery, on admission to the recovery room, and daily until postoperative day (POD) 5. The volumes of blood loss and of intraoperative fluids administered were recorded. Coagulation abnormalities observed immediately after surgery may be related mostly to blood loss and to the diluting effect of the intraoperative infused fluids, although the extent of the resection appears to be the most important factor in the extension of the PT observed from POD 1. In conclusion, significant alterations in PT and platelet values were observed in our patients who underwent uncomplicated major liver resection for living donor liver transplantation. Because the potential benefits of epidural analgesia for liver resection are undefined according to available data, additional prospective randomized studies comparing the effectiveness and safety of intravenous versus epidural analgesia in this patient population should be performed. PMID- 15350006 TI - Donor selection for procurement of right posterior segment graft in living donor liver transplantation. AB - The right posterior segment (RPS) graft was introduced to overcome graft-size mismatch when the donor liver demonstrates a disproportionately small left lobe (LL). As variants of liver anatomy seemed to be related to the feasibility of RPS graft procurement, in 2003, we performed a prospective study to assess its feasibility in 197 consecutive living donors. Variants of the portal vein (PV) were classified as type I (bifurcation), II (trifurcation), and III (independent RPS PV branching from main PV). The right hepatic artery, vein, and bile duct were also classified according to their branching pattern and location. PV variations were type I in 157 (79.7%) donors, type II in 15 (7.6%) donors, and type III in 25 (12.7%) donors. Mean volume proportion of LL plus caudate lobe was 35.3 +/- 3.8% (range 24-4) of the whole liver volume (WLV). On exclusion of donors with LL greater than 35% of WLV, there were 14 (7.1%) donors revealing RPS greater than LL by over 3% of WLV. Of these 14 donors, 3 had type I PV with artery or bile duct anatomy not suitable for RPS procurement. One donor with type II PV and 9 out of 10 donors with type III PV met the anatomical conditions for RPS graft procurement. With the exclusion of caudate lobe volume, LL volume became less than 30% of WLV in all of these 14 donors. We successfully procured 3 RPS grafts, all with type III PV, out of 197 consecutive living donors. In conclusion, successful RPS graft procurement is highly possible, only when LL is disproportionately small (<30% of WLV) and the PV variant is type III. PMID- 15350007 TI - Right lateral sector graft as a feasible option for partial liver transplantation. PMID- 15350008 TI - High hilar dissection: new technique to reduce biliary complication in living donor liver transplantation. AB - Biliary complications after living donor liver transplantation (LDLT) continue to be problematic. For reducing the biliary complications, the authors applied an intrahepatic Glissonian approach to the recipient hepatectomy. We called this Glissonian dissection technique at the high hilar level high hilar dissection (HHD). In this study, we introduced this HHD technique and evaluated its outcome in 31 recipients of a living donor liver transplant (LDLT). With total occlusion of hepatoduodenal ligament Glissonia pedicles were divided at the intrahepatic level at the third level of pedicles or beyond. After portal vein and hepatic artery were isolated from the hepatoduodenal ligament, unused bile ducts and bleeding were controlled with continuous suture of the hilar plate. Single duct anastomosis was performed in about 21 and dual duct anastomosis in 10 recipients. Bile leakage of the biliary anastomosis did not occur. There were 6 biliary complications in five patients; 2 bile leaks from the cut liver surface and 4 biliary strictures of which one of unknown etiology. In none of the patients with biliary complications, conversion to a hepaticojejunostomy was necessary. This new HHD technique during recipient hepatectomy may contribute to reduce the biliary complications in duct-to-duct anastomosis by allowing a tension free anastomosis and preserving adequate blood supply to the bile duct. Moreover, it facilitates multiple ductal anastomoses without difficult surgical manipulation. PMID- 15350009 TI - Reconstruction of isolated caudate portal vein in left liver graft. AB - The addition of the caudate lobe to a left liver graft is known to increase graft volume by 9% and has been shown to be useful for overcoming borderline graft recipient size mismatch in adult living donor liver transplantation. To assure full graft viability and regeneration, all of the feeding vessels for the caudate lobe should be preserved or reconstructed. However, more knowledge is needed about portal vein reconstruction for the caudate lobe. From January 1996 to August 2003, 238 living donor liver transplantations were performed at our institution. Of these, 67 donors underwent left hepatectomy with the caudate lobe and the middle hepatic vein. An isolated caudate portal vein originating from the left sidewall of the portal branches for the Spiegelian lobe (P1) was encountered in 9 donors (13.4%). The isolated P1 was reconstructed in 3 of the 9 cases using the pantaloon technique or interposition of an autovein graft. There were no complications related to P1 reconstruction and patency was confirmed by computerized tomography (CT) 1 month after transplantation. In the remaining 6 cases, the isolated P1 was very small (less than .5 mm) and did not require reconstruction. Isolated P1s are relatively rare, but when they are both present and large, it is advisable to undertake reconstruction that assures full graft function of the caudate lobe. PMID- 15350010 TI - Glutathione S-transferase T1 mismatch constitutes a risk factor for de novo immune hepatitis after liver transplantation. AB - A new form of autoimmune hepatitis referred to as de novo, has been reported after liver transplantation during the past 5 years. The features are identical to those of classical autoimmune hepatitis (AIH), but the facts involved in the onset and outcome of this type of graft dysfunction are still unclear. The identification of antibodies directed to glutathione S-transferase T1 (GSTT1) in the sera of patients with de novo immune hepatitis led us to the description of an alloimmune reaction due to a GSTT1 genetic incompatibility between donor and recipient. We analyzed a cohort of 110 liver transplant patients treated in the liver transplant unit of our hospital during a period of 1 year, from September 2002 to October 2003. We found the following distribution of the GSTT1 genotypes (recipient/donor): +/+ = 66, +/- = 23, -/+ = 15, -/- = 6. Six of these patients were diagnosed with de novo immune hepatitis; all of them belong to the group of negative recipients with positive donors, and all produced anti-GSTT1 antibodies. This genetic combination is associated with a statistically significant increased risk of de novo immune hepatitis (IH) in liver transplant patients (P < .0001 by the Fisher exact test). In conclusion, our results clearly establish the importance of the GSTT1 genotype from donor and recipient of a liver transplant as a predictive marker for de novo IH. At the same time, we confirmed our initial results that only this particular donor/recipient combination triggers the anti GSTT1 antibody production. PMID- 15350011 TI - Insulin in UW solution exacerbates hepatic ischemia / reperfusion injury by energy depletion through the IRS-2 / SREBP-1c pathway. AB - Ischemia / reperfusion (I / R) injury is related to tissue graft energy status. Insulin, which is currently used in the University of Wisconsin (UW) preservation solution with insulin (UWI), is an anabolic hormone and was shown to exacerbate the hepatic I / R injury in our previous study. In this study, the energy status and regulation of metabolism genes by insulin were investigated in liver grafts preserved by UW solution. Insulin could significantly decrease adenosine triphosphate (ATP) level after 3 hours of preservation, as well as total adenine nucleotides (TANs) and energy charge (EC) levels. Energy regeneration deteriorated in the grafts preserved by insulin in terms of ATP and EC levels at 24 hours after transplantation. The insulin signal was transduced through the insulin receptor substrate-2 (IRS-2) pathway and the activity of IRS-2 was decreased gradually at the messenger ribonucleic acid (mRNA) level during cold preservation. Downstream targeting genes such as sterol regulatory element binding protein-1c (SREBP-1c), glucokinase (GKC), and fatty acid synthase (FAS) genes, as well as phospho-glycogen synthase kinase-3beta (GSK-3beta) were activated and they showed the similar expression profiles during cold preservation. Lipoprotein metabolism was accelerated by insulin through upregulation of the activity of apolipoprotein C-III (Apo C-III) during cold preservation. The insulin-like growth factor-binding protein-1 pathway was inhibited during cold preservation. In conclusion, insulin in UW solution exacerbates hepatic I / R injury by energy depletion as the graft maintains its anabolic activity. The key enzyme activities of the energy-consuming process of glycogen and fatty acid synthesis as well as lipoprotein metabolism were accelerated by insulin through the IRS-2 / SREBP-1c pathway. PMID- 15350012 TI - Prospective evaluation of the role of quantitative Doppler ultrasound surveillance in liver transplantation. AB - Doppler ultrasound (DUS) is able to measure parameters of blood flow within vessels of transplanted organs, and vascular complications are associated with abnormal values. We analyzed the findings of 51 consecutive patients who underwent DUS on 2 occasions in the first postoperative week following liver transplantation for cirrhosis to determine the range of values in patients following liver transplantation. Three patients developed early vascular thromboses that were detected by the absence of a Doppler signal. In patients making an uneventful recovery, the arterial velocity tended to increase and the resistive index (RI) to decrease during the first postoperative week. All recipients were shown to have high-velocity segments within the hepatic artery, without an increase in flow resistance. Assessment of the portal vein revealed narrowing at the anastomosis, associated with a segmental doubling of flow velocity, and the mean portal venous flow decreased by approximately 20% in the first postoperative week. In conclusion, a wide range of abnormalities occurs in the vessels of liver transplant recipients, which were not associated with the development of vascular complications or affect patient management. PMID- 15350013 TI - Mycophenolate mofetil monotherapy in liver transplant recipients: a single center experience. AB - The long-term use of calcineurin inhibitors (CIs) is associated with significant morbidity in liver transplant recipients. Although mycophenolate mofetil (MMF) is well tolerated, two small studies reported an unacceptable rate of acute allograft rejection in liver transplant recipients receiving MMF monotherapy. In this study, we retrospectively investigated the safety and efficacy of MMF monotherapy in liver transplant recipients. We reviewed the medical records of all patients who underwent liver transplant at our institution. Sixteen patients were identified who received MMF either as monotherapy (n = 13) or with corticosteroids (n = 3; 2 of them for other comorbid conditions), and these patients were studied to determine the efficacy and complications. Fifteen (15/16) patients were converted from a CI to MMF because of renal insufficiency. Patients were converted to MMF monotherapy after a median of 2,056 days (range, 606-5,893) after liver transplantation. The median postconversion follow-up was 668 days (range, 60-1,509). Four patients required dialysis despite conversion; of those patients not requiring dialysis, serum creatinine stabilized and showed a trend toward improvement (2.51 +/- 1.12 mg/dL to 1.85 +/- .58 mg/dL, P = .1). However, there were 3 episodes (47, 107, and 1,203 days after conversion) of severe, irreversible allograft rejection after conversion resulting in death in 2 patients and necessitating retransplantation in 1 patient. There were no patient characteristics, except perhaps African-American race, that predicted the development of rejection. In conclusion, MMF monotherapy was associated with a significant risk (19%) of unpredictable, severe, and irreversible allograft rejection even among long-term transplant survivors. Caution should be exercised before converting patients to MMF monotherapy. PMID- 15350014 TI - Liver transplantation for hepatocellular carcinoma: further considerations on selection criteria. AB - The selection criteria in liver transplantation for HCC are a matter of debate. We reviewed our series, comparing two periods: before and after 1996, when we started to apply the Milan criteria. The study population was composed of patients with a preoperative diagnosis of HCC, confirmed by the pathological report and with a survival of >1 year. Preoperative staging as revealed by radiological imagining was distinguished from postoperative data, including the variable of tumor volume. After 1996 tumor recurrences significantly decreased (6 out of 15 cases, 40% vs. 3 out of 48, 6.3%, P < .005) and 5-year patient survival improved (42% vs. 83%, P < .005). Not meeting the Milan criteria was significantly related to higher recurrence rate (37.5% vs. 12.7%, P < .05) and to lower 5-year patient survival (38% vs. 78%, P < .005%) in the preoperative analysis, but not in the postoperative one. The alfa-fetoprotein level of more than 30 ng/dL and the preoperative tumor volume of more than 28 cm3 predicted HCC recurrences in the univariate and mutivariate analysis (P < .005 and P < .05, respectively). The ROC curve showed a linear correlation between preoperative tumor volume and HCC recurrence. Milan criteria significantly reduced tumor recurrences after liver transplantation, improving long-term survival. In conclusion, the efficacy of tumor selection criteria must be analyzed with the use of preoperative data, to avoid bias of the postoperative evaluation. Tumor volume and alfa-fetoprotein level may improve the selection of patients. PMID- 15350015 TI - Orthotopic liver transplantation for portosystemic encephalopathy in an adult with congenital absence of the portal vein. AB - Congenital absence of the portal vein (CAPV) is a very rare venous malformation in which mesenteric venous blood drains directly into the systemic circulation. There is no portal perfusion of the liver and no portal hypertension. This abnormality is usually coincidentally discovered in children, the majority of whom have no signs of encephalopathy and only slightly abnormal liver function tests. Additional anomalies common in CAPV are cardiovascular abnormalities and hepatic tumors. To date, only 5 adult patients (>18 years) with CAPV have been described, none of whom underwent liver transplantation. We describe a 45-year old man with CAPV and end-stage renal insufficiency due to focal segmental glomerulopathy, who developed therapy-resistant encephalopathy with intermittently high ammonia levels. The patient underwent a combined liver and kidney transplantation and is doing well at 2.5 years of follow-up. Histopathological examination of the native liver showed no portal vein branches in the portal tracts. In conclusion, our experience suggests that, although children with CAPV usually have no symptoms of encephalopathy, this may still develop at a later stage in adult life. When encephalopathy becomes refractory to medical therapy, orthotopic liver transplantation (OLT) can be successfully performed with restoration of normal cerebral function. PMID- 15350017 TI - Permanent access to the portal system for cellular transplantation using an implantable port device. AB - A novel application of the implantable Port-a-Cath (PAC) system is described in the context of cellular transplantation. A silicone catheter was inserted in a collateral branch of the portal vein and connected to a port device positioned subcutaneously on the left thoracic cage. This permanent vascular access allowed iterative intraportal infusions of allogenic hepatocytes without the need of repeated transhepatic catheterization of the portal vein. Using this technique, repeated infusions of cryopreserved and / or fresh hepatocytes were successfully carried out in 3 children with inborn errors of liver metabolism, with the aim of progressively providing a sufficient mass of transplanted liver cells to stabilize the metabolic condition of the patients. We suggest that this technique might also be valuable in pancreatic islet cell transplantation. PMID- 15350016 TI - Sharing the middle hepatic vein between donor and recipient: left liver graft procurement preserving a large segment VIII branch in donor. AB - There are few reported techniques to minimize the congestion in the donor after left liver graft procuring. If a large tributary of the middle hepatic vein (MHV) draining segment VIII (V8) converges into the root of the MHV in a donor of left liver, this branch should be preserved on the donor side. The volume of congested area when the V8 was ligated was predicted preoperatively by computed tomography (CT) and examined intraoperatively by the clamp test. Postoperative regeneration of the donor liver was evaluated by CT volumetry. This technique was used in 3 cases. The regeneration rate after 3 months of the right paramedian sector was 27, 38, and 8%, and that of the right lateral sector was 31, 63, and 39% in each donor, respectively. No severe complications occurred in the donors. In conclusion, V8 preservation in donors who underwent left liver resection led to satisfactory regeneration both of the right paramedian and lateral sectors and can minimize congestion in remnant liver. PMID- 15350018 TI - Cavoatrial shunt in the treatment of suprahepatic vena cava stricture after liver transplantation. PMID- 15350019 TI - Can liver transplantation improve our understanding of the pathophysiology of iron overload? PMID- 15350020 TI - Ascites after transplantation--a mystery. PMID- 15350021 TI - Characteristics of the anterolateral thigh flap in a Western population and its application in head and neck reconstruction. AB - BACKGROUND: Although the anterolateral thigh flap has been extensively used for head and neck reconstruction in Asia, reported variations of vascular anatomy seem confusing and may have contributed to the unpopularity of this flap in the United States. The purposes of this study are to classify the vascular anatomy and to assess the suitability of this flap for head and neck reconstruction in a Western population. METHODS: Seventy-two consecutive anterolateral thigh flaps for head and neck reconstruction was retrospectively reviewed. RESULTS: The number of cutaneous perforators for the anterolateral thigh flap ranged from one to three. On the basis of their location and origin, a simple classification system is introduced to assist flap dissection. Of the 72 thighs explored, 68 flaps (94%) were raised successfully. CONCLUSIONS: The vascular anatomy of the flap follows predictable patterns. The anterolateral thigh flap is well suited for head and neck reconstruction in Westerners. PMID- 15350022 TI - Secondary end-to-end repair of extensive facial nerve defects: surgical technique and postoperative functional results. AB - BACKGROUND: Repair of the transected facial nerve is imperative for restoration of muscle function, including the ability to produce appropriate facial expressions. Injury might involve the main trunk and its several branches. Restoration of function presupposes meticulous repair of all injured nerve branches. METHODS: Here we report three cases of secondary tension-free end-to end coaptation of a transected trunk and branches of the facial nerve by removal of the superficial part of the parotid gland. RESULTS: Facial tone and symmetry at rest and motion were achieved. In two patients, a slight residual synkinesis is observed under stress. CONCLUSIONS: Direct end-to-end coaptation of the facial nerve and its branches by the technique described should be considered before deciding on grafts or rerouting procedures to deal with gaps of up to 15 mm. This technique is not recommended in the presence of infection and nerve defects. Intensive postoperative physiotherapy is required for optimal results. PMID- 15350023 TI - High frequency of HPV16-associated head and neck squamous cell carcinoma in the Puerto Rican population. AB - BACKGROUND: Recent evidence has accumulated suggesting that human papillomavirus (HPV) plays a role in the development of head and neck squamous cell carcinoma (HNSCC). HPV16 is the most common of the HPV subtypes associated with oral and laryngeal malignancies. This study estimated the prevalence of HPV16 DNA in Puerto Rican patients with HNSCC. METHODS: DNA was extracted from frozen tissue of 118 HNSCCs. Genomic DNA was screened for the presence of HPV16 DNA with E6 specific and E7-specific primers. RESULTS: HPV16 was detected in tumor tissue of 52 patients (44%) with HNSCC. The oropharynx had a slightly higher incidence of HPV16 DNA. Fifteen of 66 patients with HPV16-negative HNSCC later had recurrences. Positivity for HPV16 was independent of the tumor grade, tumor stage, nodal status, and tobacco or alcohol use. The 3-year survival rate was higher in HPV16-positive patients than in HPV16-negative patients (36% vs 21%). CONCLUSIONS: Our findings suggest that HPV16 may play a role in the etiology of a subgroup of HNSCC in Puerto Ricans. Overall survival times of the HPV16-positive patients were not significantly different from those of HPV16-negative patients. Increasing our understanding of the role of HPV16 in the etiology of HNSCC might facilitate the development of new treatment modalities for this subgroup of HNSCC. PMID- 15350024 TI - Morbidity of the neck after head and neck cancer therapy. AB - BACKGROUND: Studies on morbidity of the neck after head and neck cancer therapy are scarcely described. METHODS: Patients who underwent surgery, including neck dissection, with and without radiation therapy at least 1 year before the study were asked to participate. We assessed neck pain, loss of sensation, range of motion of the cervical spine, and shoulder pain. RESULTS: Of the 220 patients who were invited, 153 (70%) participated in the study. Neck pain was present in 33% of the patients (n = 51), and shoulder pain was present in 37% of the patients (n = 57). Neuropathic pain of the neck was present in 32% (n = 49); myofascial pain, in 46% (n = 70); and joint pain, in 24% (n = 37). Loss of sensation of the neck was present in 65% (n = 99) and was related to type of neck dissection and radiation therapy. Range of motion of the neck was significantly decreased because of the neck dissection and/or radiation therapy in lateral flexion away from the operated side. CONCLUSIONS: The occurrences of morbidity of the neck after cancer therapy were considerable and consisted of neck pain, loss of sensation, and decreased range of motion. PMID- 15350025 TI - Incidence and disease-free survival after surgical therapy of central giant cell granulomas of the jaw in The Netherlands: 1990-1995. AB - BACKGROUND: The central giant cell granuloma (CGCG) of the jaw is a rare benign tumor with an unknown etiology. Epidemiologic data on CGCGs in a general population are not available, nor are data on the overall outcome of surgical therapy. In this article, an overview of these aspects in the Dutch population from January 1, 1990, until January 1, 1995, is presented. METHODS: All CGCGs that were diagnosed and surgically treated in The Netherlands from 1990 until 1995 were evaluated. RESULTS: The incidence is 0.00011%, and the disease-free survival after surgical therapy after 5 years is 76.1%. There is a significantly higher recurrence rate in young males, which cannot be explained by clinical signs and symptoms of the lesion. CONCLUSION: The CGCG is a rare affliction with a high recurrence rate after surgical therapy, especially in young males. PMID- 15350026 TI - Radiation-induced xerostomia in patients with head and neck cancer: pathogenesis, impact on quality of life, and management. AB - BACKGROUND: Xerostomia is a common, debilitating complication of radiation therapy (RT) for head and neck cancer. This article reviews the pathogenesis of radiation-induced xerostomia, its impact on quality of life (QOL), and treatment options. METHODS: Virtually all patients undergoing RT for head and neck cancers have xerostomia, which causes oral discomfort and pain, increased dental caries and oral infection, and difficulty speaking and swallowing. This significantly impairs QOL and can compromise nutritional intake and continuity of cancer therapy. The literature describing pathogenesis, impact on QOL of radiation induced xerostomia, and preventive and interventional therapies was reviewed. RESULTS: Current management strategies include stringent dental and oral hygiene; parotid-sparing radiation techniques to prevent or minimize xerostomia; and pharmacotherapies, such as salivary substitutes and sialogogues. Future strategies may include advanced three-dimensional intensity-modulated RT techniques, salivary gland transfer, newer sialogogues, and gene therapy. CONCLUSIONS: New treatment approaches to xerostomia from RT for head and neck cancer may result in significant improvement in patient QOL. PMID- 15350027 TI - Reconstruction of the hard palate using the radial forearm free flap: indications and outcomes. AB - BACKGROUND: Although prosthetic obturation is the "gold standard" for restoration of hard-palate defects, obturators can be problematic. We present 10 cases of palatal reconstruction with the radial forearm free flap and compare patient satisfaction with defect-matched patients rehabilitated with prosthetic obturation. METHODS: Twelve patients who underwent radial forearm free flap (RFFF) reconstruction of a hard-palate defect and eight patients, with similar sized defects who were rehabilitated with a prosthetic obturator, were evaluated for donor site and recipient site complications, diet, and patient satisfaction. RESULTS: All the patients in both groups were able to resume an unrestricted diet with normal mastication and articulation. Both groups achieved equivalent satisfaction scores with regard to appearance, chewing, and taste; however, the patients reconstructed with an RFFF reported higher satisfaction scores in speech, comfort, convenience, and social interaction. CONCLUSIONS: RFFF reconstruction of hard-palate defects provides a functional alternative to conventional prosthetic obturators. PMID- 15350028 TI - Correlation of Epstein-Barr virus DNA in cell-free plasma, functional imaging and clinical course in locally advanced nasopharyngeal cancer: a pilot study. AB - BACKGROUND: We evaluated the feasibility of using quantitative real-time polymerase chain reaction (PCR) in monitoring Epstein-Barr virus (EBV) DNA concentrations in cell-free plasma of patients with localized nonmetastatic nasopharyngeal carcinoma (NPC) treated with chemoradiation. METHODS: Cell-free plasma EBV DNA was quantified in six patients with locally advanced, nonmetastatic nasopharyngeal cancer (NPC) who underwent chemoradiation therapy (CRT) and correlated with magnetic resonance imaging (MRI), positron emission tomography (PET) scans, and clinical course. RESULTS: The mean concentration of EBV DNA was 24,610 copies/mL, 682 copies/mL, and 64.5 copies/mL in the pretreatment, posttreatment, and last follow-up samples, respectively. Four of the six patients had normal PET scans and reduction of EBV DNA copy numbers to minimal levels after treatment and, despite equivocal posttreatment MRI scans, are clinically free of recurrent disease. Two of the six patients had elevated EBV DNA copy numbers immediately after treatment, but a metastatic workup was clear at the time; both subsequently relapsed with distant metastases 5 and 11 months later. Although the posttreatment PET scans were abnormal in both of these cases, they were not predictive of the subsequent sites of tumor relapse. CONCLUSIONS: To our knowledge, this is the first report on the correlation between EBV DNA levels and PET scan results in patients with NPC. Reductions in EBV DNA levels and normal PET scans after treatment were consistent among patients who had residual abnormality on MRI images. Persistently elevated EBV DNA levels and abnormal PET scans after treatment suggest residual disease. Further evaluations are required to determine the relative contributions of these two novel techniques in predicting residual disease in locally advanced NPC. PMID- 15350029 TI - Efficacy of piperacillin-tazobactam in the treatment of surgical wound infection after clean-contaminated head and neck oncologic surgery. AB - BACKGROUND: Although perioperative antibiotic prophylaxis has significantly reduced surgical wound infection rates, this complication is still a frequent complication of head and neck cancer surgery. Because these infections are typically polymicrobial, our study evaluated the safety and efficacy of piperacillin-tazobactam in the treatment of surgical wound infection after clean contaminated head and neck oncologic surgery. METHODS: In this multicenter, prospective clinical trial, 70 patients with surgical wound infection received piperacillin-tazobactam. RESULTS: Of patients who were evaluable, 92.4% were also clinically cured or improved, and the bacteriologic eradication rate was 80.3%. Of the 70 patients enrolled in the study, six (8.5%) experienced six adverse events: two cases of moderate diarrhea, one allergic skin reaction, and three cases of phlebitis. No deaths were attributable to the study drug. CONCLUSIONS: Piperacillin-tazobactam is a good choice of treatment as monotherapy for surgical wound infection after clean-contaminated head and neck oncologic surgery. PMID- 15350030 TI - Imatinib mesylate as treatment for adenoid cystic carcinoma of the salivary glands: report of two successfully treated cases. AB - BACKGROUND: Adenoid cystic carcinoma (ACC) is a malignant neoplasia of the salivary glands that is treated primarily by surgery. Local control and survival are usually compromised despite surgery. Expression of KIT tyrosine kinase is involved in the pathogenesis of ACC. Imatinib mesylate is a potent inhibitor of KIT tyrosine kinase, so we explored the possibility that ACC could be a potential target for this drug. METHODS: We report two cases of unresectable ACC treated with imatinib mesylate in the context of recurrent disease (case 1) and locally advanced tumor at its initial presentation (case 2). RESULTS: Both patients responded well to treatment with imatinib mesylate. Significant regression of recurrent disease (case 1) resulted in a successful salvage surgical resection; the locally advanced tumor (case 2) had an excellent response to treatment, but, unfortunately, the patient refused salvage resection. CONCLUSION: This is the first time ACC is reported to respond to imatinib mesylate. Studies in which more patients are enrolled in controlled clinical trials are needed to confirm this observation. PMID- 15350032 TI - Neurological manifestation of sacral tumors. AB - An extensive analysis of the existing literature concerning sacral tumors was conducted to characterize their clinical manifestations. Although certain specific manifestations can be attributed to some of the tumor types, a more general pattern of clinical presentation of an expansive sacral lesion can be elaborated. Local pain with or without pseudoradicular or radicular radiation is the most frequent initial symptom and is usually followed by the manifestation of a lumbosacral sensorimotor deficit; bladder/bowel and/or sexual dysfunction appear throughout the natural course of disease. PMID- 15350033 TI - Imaging of sacral tumors. AB - Sacral tumors are somewhat unique to the skeletal system in that the prevalence of various neoplasms is different from that at other osseous sites. There are also unique imaging-related considerations, related in part to diagnosis, but particularly to the evaluation site for tumor biopsy sampling and resection. In this report, the authors describe imaging features of the most common sacral lesions, provide numerous radiological examples, and give suggestions for optimal imaging of this region of the body. PMID- 15350034 TI - Anatomy of the sacrum. AB - One of the basic tenets of performing surgery is knowledge of the relevant anatomy. Surgeons incorporate this knowledge along with factors, such as biomechanics and physiology, to develop their operative approaches and procedures. In the diagnosis and management of sacral tumors, the need to be familiar with the anatomy of the sacrum is no less important than knowledge of the pathological entity involved. This article will provide an overview of the embryology and anatomy of the sacrum, along with concepts as applied to surgical intervention. PMID- 15350035 TI - Embolization of sacral tumors. AB - The management of sacral tumors is challenging because of difficulties in accessing the lesion, the high rate of local recurrence, extensive vascularity causing significant intraoperative blood loss, resistance to radiation therapy, and risk of malignant transformation. Although surgery is the main treatment for many sacral tumors, embolization is a valuable primary and adjunctive therapy. Patients with benign lesions, including aneurysmal bone cysts and giant cell tumors, have responded to embolization with resolution of their symptoms and with ossification of their lesions. Embolization is used as a primary therapy for metastatic lesions and results in neurological improvement, reduced tumor size, and decreased spinal canal compromise. It is also used as an adjuvant therapy to reduce intraoperative blood loss and to aid in the resection of benign, malignant, and metastatic sacral lesions. It is important to note that embolization techniques are a valuable resource in the treatment of sacral tumors, and, overall, embolization should always be considered in patients with sacral tumors. PMID- 15350036 TI - Surgical techniques for total sacrectomy and spinopelvic reconstruction. AB - The surgical management of sacral tumors requires partial or total sacrectomy and spinopelvic reconstruction. These lesions present a great surgical challenge, because most spine surgeons are unfamiliar with the techniques required for these procedures. The authors describe a step-by-step operative technique and provide several illustrations. Total sacrectomy is performed by sequential anterior and posterior approaches that involve a rectus abdominis pullthrough pedicle flap reconstruction. The anterior procedure is an intraperitoneal approach used to expose the anterior aspect of the tumor, to ligate the main tumor vessels, and to conduct an anterior partial sacrectomy. After this, the rectus abdominis myocutaneous flap, based on the inferior epigastric vessel, is prepared, and a posterior sacrectomy is performed, dividing all sacral nerve roots in the thecal sac. After complete en bloc extirpation of the sacrum with tumor, spinopelvic reconstruction and closure with a myocutaneous flap are performed. Spinopelvic reconstruction is undertaken using a modified Galveston technique or double iliac screw fixation combined with posterior lumbar segmental fixation. These provide a long lever arm within the ilium to counteract the forces exerted by the lumbar spine. Understanding the nature of the disease as well as the biomechanics of the lumbosacral pelvic area and spinopelvic fixation will help surgeons select the appropriate treatment for sacral tumors. PMID- 15350037 TI - Lumbosacropelvic junction reconstruction resulting in early ambulation for patients with lumbosacral neoplasms or osteomyelitis. AB - Lumbosacropelvic junction instability may result from a variety of disease processes including primary and metastatic sacral tumors and degenerative disease. Regardless of the origin of the disease, restoring or maintaining spinal stability at this junction is essential for normal translation of axial forces from the lumbar spine and sacrum to the pelvis. Spinal stability is also critical for maintaining structural integrity, preventing neurological function deterioration, and alleviating resultant mechanical or axial pain. In this report, the authors describe one option for safe and effective spinal pelvic stabilization by using a transiliac rod and iliac bolt construct, which results in early postoperative ambulation, preserved neurological function, and reduced axial pain in selected patients. PMID- 15350038 TI - CyberKnife frameless single-fraction stereotactic radiosurgery for tumors of the sacrum. AB - OBJECT: The role of stereotactic radiosurgery for the treatment of intracranial lesions is well established. The experience with radiosurgery for the treatment of spinal and sacral lesions is more limited. Sacral lesions should be amenable to radiosurgical treatment similar to that used for their intracranial counterparts. The authors evaluated a single- fraction radiosurgical technique performed using the CyberKnife Real-Time Image-Guided Radiosurgery System for the treatment of the sacral lesion. METHODS: The CyberKnife is a frameless radiosurgery system based on the coupling of an orthogonal pair of x-ray cameras to a dynamically manipulated robot-mounted linear accelerator possessing six degrees of freedom, which guides the therapy beam to the intended target without the need for frame-based fixation. All sacral lesions were located and tracked for radiation delivery relative to fiducial bone markers placed percutaneously. Eighteen patients were treated with single-fraction radiosurgery. Tumor histology included one benign and 17 malignant tumors. Dose plans were calculated based on computerized tomography scans acquired using 1.25-mm slices. Planning treatment volume was defined as the radiographically documented tumor volume with no margin. Tumor dose was maintained at 12 to 20 Gy to the 80% isodose line (mean 15 Gy). Tumor volume ranged from 23.6 to 187.4 ml (mean 90 ml). The volume of the cauda equina receiving greater than 8 Gy ranged from 0 to 1 ml (mean 0.1 ml). All patients underwent the procedure in an outpatient setting. No acute radiation toxicity or new neurological deficits occurred during the mean follow-up period of 6 months. Pain improved in all 13 patients who were symptomatic prior to treatment. No tumor progression has been documented on follow-up imaging. CONCLUSIONS: Stereotactic radiosurgery was found to be feasible, safe, and effective for the treatment of both benign and malignant sacral lesions. The major potential benefits of radiosurgical ablation of sacral lesions are relatively short treatment time in an outpatient setting and minimal or no side effects. This new technique offers a new and important therapeutic modality for the primary treatment of a variety of sacral tumors or for lesions not amenable to open surgical techniques. PMID- 15350039 TI - Radiosurgery and radiotherapy for sacral tumors. AB - Sacral tumors represent a small subset of spinal lesions and typically include chordomas, metastases, other primary bone tumors, and benign schwannomas. Resection is the standard treatment for many sacral tumors, but many types of sacral lesions have the potential for recurrence after excision. In these cases, adjuvant radiotherapy is often beneficial. Although conventional radiotherapy plays an important role in the management of spinal lesions, the radiation doses required for adequate local control of many sacral lesions generally exceed the tolerance doses of normal tissues, thus limiting its definitive role in the management of sacral tumors. Recent advances in the field of stereotactic radiosurgery have allowed precise targeting of the sacrum. In this report the authors review the use of these two forms of radiation treatment and their role in managing sacral tumors. PMID- 15350040 TI - Current management of sacral chordoma. AB - Sacral chordomas are relatively rare, locally invasive, malignant neoplasms. Although metastasis is infrequent at presentation, the prognosis for patients with chordoma of the sacrum is reported to be poor and attributable in most cases to intralesional resection. The value of adjuvant treatment is uncertain, and resection remains the primary mode of treatment. Chordomas are difficult to excise completely, but recent improvements in imaging and surgical techniques have allowed surgeons to perform more frequently en bloc sacral resections with wide surgical margins. The technical challenges of such operations, and the functional costs for the patient (with respect to anorectal and urogenital dysfunction) are significantly increased when the tumor involves high sacral levels. The authors review the clinical presentation and natural history of sacral chordoma and discuss the current treatment techniques and outcomes. PMID- 15350041 TI - Diffuse large B-cell lymphoma presenting as a sacral tumor. Report of two cases. AB - Primary lymphomas of the sacrum are rare tumors, reported only in a few cases in the literature. The authors describe two patients with diffuse large B-cell lymphomas presenting as a sacral tumor. In the first case a 52-year-old man presented with progressive back pain, bilateral radicular pain, and saddle block anesthesia secondary to a lytic, expansile soft-tissue mass. The mass arose from the sacrum and eroded through the right S-1 to S-4 foramina and extended into the epidural space of the spinal canal. On magnetic resonance imaging, the sacral mass enhanced homogeneously with Gd. In the second case a 64-year-old man presented with left-sided radicular pain, paresthesias, and progressive weakness due to a lytic soft-tissue mass in the left sacral ala extending into the left L 5 and S-1 foramina. Metastatic workup in each patient demonstrated unremarkable findings. In both cases, an open biopsy procedure was performed after nondiagnostic examination of needle biopsy samples. Histopathological examination showed evidence consistent with diffuse large B-cell lymphoma in both patients. In the first case the disease was classified as Stage IAE, and the patient subsequently underwent four cycles of cyclophosphamide/doxorubicin/vincristine/prednisone (CHOP)- and rituximab-based chemotherapy followed by consolidation radiotherapy. In the second case the disease was also classified as Stage IAE, and the patient underwent CHOP-based chemotherapy and consolidation radiotherapy. In both cases radiography demonstrated a decrease in size of the sacral lymphomas. The authors review the clinical, radiological, and histological features of sacral lymphomas. Lymphoma should be considered in the differential diagnosis of sacral tumors. PMID- 15350042 TI - Sacral hemangioblastoma in a patient with von Hippel-Lindau disease. Case report and review of the literature. AB - Hemangioblastomas are histologically benign neoplasms that occur sporadically or as part of von Hippel-Lindau disease. Hemangioblastomas may occur anywhere along the neuraxis, but sacral hemangioblastomas are extremely rare. To identify features that will help guide the operative and clinical management of these lesions, the authors describe the management of a large von Hippel-Lindau disease associated sacral hemangioblastoma and review the literature. The authors present the case of a 38-year-old woman with von Hippel-Lindau disease and a 10-year history of progressive back pain, as well as left lower-extremity pain and numbness. Neurological examination revealed decreased sensation in the left S-1 and S-2 dermatomes. Magnetic resonance imaging demonstrated a large enhancing lesion in the sacral region, with associated erosion of the sacrum. The patient underwent arteriography and embolization of the tumor and then resection. The histopathological diagnosis was consistent with hemangioblastoma and showed intrafascicular tumor infiltration of the S-2 nerve root. At 1-year follow-up examination, pain had resolved and numbness improved. Sacral nerve root hemangioblastomas may be safely removed in most patients, resulting in stabilization or improvement in symptomatology. Generally, hemangioblastomas of the sacral nerve roots should be removed when they cause symptoms. Because they originate from the nerve root, the nerve root from which the hemangioblastoma originates must be sacrificed to achieve complete resection. PMID- 15350043 TI - Nerve sheath tumors involving the sacrum. Case report and classification scheme. AB - Nerve sheath tumors that involve the sacrum are rare. Delayed presentation is common because of their slow-growing nature, the permissive surrounding anatomical environment, and nonspecific symptoms. Consequently, these tumors are usually of considerable size at the time of diagnosis. The authors discuss a case of a sacral nerve sheath tumor. They also propose a classification scheme for these tumors based on their location with respect to the sacrum into three types (Types I-III). Type I tumors are confined to the sacrum; Type II originate within the sacrum but then locally metastasize through the anterior and posterior sacral walls into the presacral and subcutaneous spaces, respectively; and Type III are located primarily in the presacral/retroperitoneal area. The overwhelming majority of sacral nerve sheath tumors are schwannomas. Neurofibromas and malignant nerve sheath tumors are exceedingly rare. Regardless of their histological features, the goal of treatment is complete excision. Adjuvant radiotherapy may be used in patients in whom resection was subtotal. Approaches to the sacrum can generally be classified as anterior or posterior. Type I tumors may be resected via a posterior approach alone, Type III may require an anterior approach, and Type II tumors usually require combined anterior-posterior surgery. PMID- 15350044 TI - Giant cell tumor of the sacrum. AB - Giant cell tumor (GCT) is a locally highly aggressive tumor of bone comprising 5 to 10% of all benign bone tumors. The sacrum is the third most common site of involvement. Patients with sacral GCTs present with localized pain in the lower back that may radiate to one or both lower limbs. Vague abdominal complaints and bowel and bladder symptoms may also be present. Neuroimaging workup should include advanced modalities, preferably magnetic resonance imaging, prior to obtaining a biopsy specimen. Giant cell tumor has a 1 to 5% rate of metastasizing to the lung and may convert to a fulminate malignant variant, which has a very poor prognosis. The standard treatment for GCT is curettage combined with adjuvant bone grafting or cement-augmented stabilization. In appropriately selected cases, sacral resection is a valuable procedure to effect local tumor control and overall survival. Embolization may also prove palliative and/or curative in cases in which the tumor is unresectable or refractory to treatment. PMID- 15350045 TI - Benign sacral tumors. AB - Primary tumors of the sacrum are rare. In adults, the most common sacral tumors are metastases. The most common primary sacral tumor is a chordoma. Chordomas along as well as tumors such as chondrosarcomas, osteosarcomas, myxopapillary ependymomas, myelomas, and Ewing sarcomas are considered malignant. In this article the authors focus on benign sacral tumors. PMID- 15350046 TI - Diagnosis and management of sacral Tarlov cysts. Case report and review of the literature. AB - Perineurial (Tarlov) cysts are meningeal dilations of the posterior spinal nerve root sheath that most often affect sacral roots and can cause a progressive painful radiculopathy. Tarlov cysts are most commonly diagnosed by lumbosacral magnetic resonance imaging and can often be demonstrated by computerized tomography myelography to communicate with the spinal subarachnoid space. The cyst can enlarge via a net inflow of cerebrospinal fluid, eventually causing symptoms by distorting, compressing, or stretching adjacent nerve roots. It is generally agreed that asymptomatic Tarlov cysts do not require treatment. When symptomatic, the potential surgery-related benefit and the specific surgical intervention remain controversial. The authors describe the clinical presentation, treatment, and results of surgical cyst fenestration, partial cyst wall resection, and myofascial flap repair and closure in a case of a symptomatic sacral Tarlov cyst. They review the medical literature, describe various theories on the origin and pathogenesis of Tarlov cysts, and assess alternative treatment strategies. PMID- 15350048 TI - Magnetically alignable phase of phospholipid "bicelle" mixtures is a chiral nematic made up of wormlike micelles. AB - We have studied the phase behavior of binary mixtures of long- and short-chain lipids, namely, dimyristoyl phosphatidylcholine (DMPC) and dihexanoyl phosphatidylcholine (DHPC), using optical microscopy and small-angle neutron scattering. Samples with a total lipid content of 25 wt %, corresponding to ratios Q ([DMPC]/[DHPC]) of 5, 3.2, and 2, are found to exhibit an isotropic (I) -> chiral nematic (N) --> lamellar phase sequence on increasing temperature. The I-N transition coincides with the chain melting transition of DMPC at Q = 5 and 3.2, but the N phase forms at a higher temperature for Q = 2. All three samples form multilamellar vesicles in the lamellar phase. Our results show that disklike "bicellar" aggregates occur only in the lower temperature isotropic phase and not in the higher temperature magnetically alignable N phase, where they were previously believed to exist. The N phase is found to consist of long, flexible wormlike micelles, their entanglement resulting in the very high viscosity of this phase. PMID- 15350047 TI - Inverted colloidal crystals as three-dimensional cell scaffolds. AB - A new type of three-dimensional scaffold with inverted colloidal crystal geometry for the investigation of topological effects in cell cultures is introduced in this publication. The scaffolds are made by infiltration of the hexagonal crystal lattice of polystyrene spheres with sol-gel formulation and subsequent annealing. It possesses a relatively high degree of order among existing cell scaffolds and affords tight control over the scaffold porosity and tissue organization. The prepared scaffolds can be a convenient system for the investigation of cell-cell and cell-matrix interactions. Their biocompatibility is demonstrated for human hepatocellular carcinoma HEP G2 and human bone marrow HS-5 cell cultures. A preliminary effect of the scaffold topology on cell proliferation is observed. HEP G2 hepatocytes form a large number of 10-15 cell colonies on scaffolds made from 75-microm spheres, while their number diminishes for scaffolds from 10- and 160-microm spheres. Under similar conditions, HS-5 forms smaller colonies consisting of three to four cells in 90-microm cavities. PMID- 15350049 TI - Drastic enhancement of TiO2-photocatalyzed reduction of nitrobenzene by loading Ag clusters. AB - Ag clusters (mean diameter = 1.5 nm, standard deviation = 0.37 nm) were photodeposited on TiO(2) particles in a highly dispersed state. The loading of a small amount of the Ag clusters (0.24 wt %) dramatically enhanced both the activity for the TiO(2) photocatalytic reduction of nitrobenzene and the product selectivity of aniline. The essential action mechanism of the Ag clusters is discussed. PMID- 15350050 TI - Controlled immobilization of membrane proteins to surfaces for fourier transform infrared investigations. AB - We show that it is possible to immobilize membrane proteins uniformly and reversibly as self-assembled (sub)monolayers on nitrilotriacetic acid-covered sensor surfaces via hexahistidine sequences present either in the protein or in lipid membranes. Fourier transform infrared spectra of such self-assembled (sub)monolayers deliver important structural information of the membrane proteins and are suited to screen the function of cellular receptors. PMID- 15350051 TI - Biomimetic formation of silica thin films by surface-initiated polymerization of 2-(dimethylamino)ethyl methacrylate and silicic acid. AB - Biosilicification in diatoms is achieved by specific interactions between silaffins, composed of polypeptides and long-chain polyamines, and silicic acid derivatives. The polycondensation of silicic acids is reported to be catalyzed by the long-chain polyamines that mainly contain tertiary N-methylpropyleneimine moieties. In this report, we utilized a tertiary amine-containing polymer, poly(2 (dimethylamino)ethyl methacrylate) (poly(DMAEMA)), as a surface-grafted, biomimetic counterpart of the long-chain polyamines in silaffins and demonstrated that the surface-initiated polycondensation of silicic acids, leading to the formation of silica thin films, proceeded smoothly on surfaces presenting poly(DMAEMA), where poly(DMAEMA) was grown from gold surfaces by surface initiated, atom transfer radical polymerization. The formed silica film was characterized by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, atomic force microscopy, and scanning electron microscopy. PMID- 15350052 TI - Organogelation of diacetylene cholesteryl esters having two urethane linkages and their photopolymerization in the gel state. AB - Various diacetylene cholesteryl esters having two urethane linkages were synthesized to study the relationship between their gelation properties and chemical structures. Most of these compounds form organogels in cyclohexane, and some compounds gelatinized hexane, diethyl ether, N,N-dimethylformamide, and ethanol. The cholesteryl moieties play an important role in gel formation, but IR spectroscopic measurements show that the main driving force for gelation is hydrogen bonding of the urethane groups. Upon UV irradiation, most of the gels polymerized to give polydiacetylenes, with concomitant changes from colorless to a variety of hues, such as dark blue, orange, and pink. The polymerization proceeds efficiently in cases where the gels change color to dark blue. The polymerization reached 52% chemical yield, with the quantum yield estimated to be at least 54. Solid-state NMR spectroscopy confirmed that polymerization in the gel state proceeds via 1,4-addition. PMID- 15350053 TI - Structure of small actin-containing liposomes probed by atomic force microscopy: effect of actin concentration & liposome size. AB - Actin-containing liposomes were prepared via extrusion through 400 and 600 nm pore diameter membranes at different monomeric actin concentrations in low ionic strength buffer (G-buffer). After subjecting the liposome dispersions to high ionic strength polymerization buffer (F-buffer), topological changes in liposome structure were studied using atomic force microscopy (AFM). Paired dumbbell, horseshoelike, and disklike assemblies were observed for actin-containing liposomes extruded through 400 and 600 nm pore diameter membranes. The topology of actin-containing liposomes was found to be highly dependent on both liposome size and actin concentration. At 1 mg/mL actin, the actin-containing liposomes transformed into a disklike shape, whereas, at 5 mg/mL actin, the actin containing liposomes retained a spherical shape. On the basis of these observations, we hypothesize that actin could either polymerize on the surface of the inner leaflet of the liposome membrane or polymerize in the aqueous core of the liposome. We explain the associated shape changes induced in actin-containing liposomes on the basis of the hypothesized mechanism of actin polymerization inside the liposomes. At higher actin concentrations (5 mg/mL), we observed membrane-induced actin self-assembly in G-buffer, which implies that G-actin is able to interact directly with lipid bilayers at sufficiently high concentrations. PMID- 15350054 TI - Supramolecular assemblies of a naturally derived sophorolipid. AB - Acidic sophorolipid (SL) molecules derived from yeasts represent a novel type of asymmetrical bolaamphiphiles due to their unique structural features that include an asymmetrical polar head size (disaccharide vs COOH), a kinked hydrophobic core (cis-9-octadecenoic chain), and a non-amide polar-nonpolar linkage. Light microscopy, small- and wide-angle X-ray scattering, FT-IR spectroscopy, and dynamic laser light scattering were used to investigate the supramolecular structures of the self-assembled aggregates of SL molecules at different pH values. In acidic conditions (pH < 5.5), giant twisted and helical ribbons of 5 11 microm width and several hundreds of micrometers length were observed for the first time. Increase in solution pH values slowed ribbon formation, decreased ribbon yield, and increased the helicity and entanglements of the giant ribbons. An interdigitated lamellar packing model of acidic SL-COOH molecules with a long period of 2.78 nm, stabilized by both the strong hydrophobic association between the cis-9-octadecenoic chains and strong disaccharide-disaccharide hydrogen bonding, is proposed. The neutralization of SL-COOH in water to SL-COONa produced clear solutions with the formation of short-range ordered aggregates. At concentrations below 1.0 mg/mL, the size of self-assembled aggregates increased as the concentration increased. At concentrations above 1.0 mg/mL, narrowly distributed micellar aggregates with a constant hydrodynamic radius (R(h)) of about 100 nm are formed. The large micelles show strong angular dependence with the fast mode appearing at scattering angle theta >/= 60 degrees. PMID- 15350055 TI - Binding of dodecyltrimethylammonium bromide to pH-responsive nanocolloids containing cross-linked methacrylic acid-ethyl acrylate copolymers. AB - The binding of dodecyltrimethylammonium bromide (DoTab) to cross-linked methacrylic acid-ethyl acrylate (MAA-EA) copolymers with various MAA/EA molar ratios at different degrees of neutralization (alpha) was quantitatively studied using isothermal titration calorimetry, dynamic light scattering, surfactant selective electrode, and electrophoresis techniques. The surfactant binds to the polymers at all degrees of neutralization, but via different mechanisms. When alpha is sufficiently high, the binding is primarily electrostatic interaction between the surfactant and ionized polymer chains, which is reinforced by the micellization of electrostatically bound surfactant molecules. The saturation takes place at charge ratio ([DoTa(+)]/[ approximately COO(-)]) close to 1, indicating that the binding is a one-to-one charge neutralization between the cationic surfactant headgroups and anionic carboxylate sites of the polymers. When alpha is low, the binding of DoTab to the unneutralized polymers is driven by the hydrophobic interaction. The onset of hydrophobic binding takes place at DoTab concentration as low as 0.01 mM in 0.05 wt % polymer solution, where the saturation occurs at C(DoTab) approximately 0.19 mM and the amount of bound surfactant is approximately 0.09 mmol of DoTab/(g of polymer) at saturation concentration. The binding results in the formation of the polymer-surfactant complex. For the polymer with low MAA/EA molar ratio, the complex coagulates at a higher DoTab concentration that leads to phase separation; however, for polymers with high MAA/EA molar ratio, the complex remains dispersed and the mixture is stable even at high DoTab concentration. PMID- 15350056 TI - Thermodynamics of micellization of multiheaded single-chain cationic surfactants. AB - The energetics of micelle formation of three single-chain cationic surfactants bearing single (h = 1), double (h = 2), and triple (h = 3) trimethylammonium [(+)N(CH(3))(3)] headgroups have been investigated by microcalorimetry. The results were compared with the microcalorimetric data obtained from well-known cationic surfactant, cetyl trimethylammonium bromide (CTAB), bearing a single chain and single headgroup. The critical micellar concentrations (cmc's) and the degrees of counterion dissociation (alpha) of micelles of these surfactants were also determined by conductometry. The cmc and the alpha values increased with the increase in the number of headgroups of the surfactant. The relationship between the cmc of the surfactant in solution and its free energy of micellization (DeltaG(m)) was derived for each surfactant. Exothermic enthalpies of micellization (DeltaH(m)) and positive entropies of micellization (DeltaS(m)) were observed for all the surfactants. Negative DeltaH(m) values increased from CTAB to h = 1 to h = 2 and decreased for h = 3 whereas DeltaS(m) values decreased with increase in the number of headgroups. The DeltaG(m) values progressively became less negative with the increase in the number of headgroups. This implies that micelle formation becomes progressively less favorable as more headgroups are incorporated in the surfactant. From the steady-state fluorescence measurements using pyrene as a probe, the micropolarities sensed by the probe inside various micelles were determined. These studies suggest that the micelles are more hydrated with multiheaded surfactants and the micropolarity of micelles increases with the increase in the number of headgroups. PMID- 15350058 TI - Electrophoresis in a Carreau fluid at arbitrary zeta potentials. AB - The electrophoresis of colloidal particles has been studied extensively in the past. Relevant analyses, however, are focused mainly on the electrophoretic behavior of a particle in a Newtonian fluid. Recent advances in science and technology suggest that the electrophoresis conducted in a non-Newtonian fluid can play a role in practice. Here, the electrophoresis of a concentrated colloidal dispersion in a Carreau fluid is investigated under the conditions of arbitrary electrical potential where the effect of double-layer polarization may be significant. A pseudo-spectral method coupled with a Newton-Raphson iteration scheme is used to solve the governing equations, which describe the electric, the flow, and the concentration fields. The results of numerical simulation reveal that, due to the effect of shear thinning, the electrophoretic mobility for the case of a Carreau fluid is greater than for that of a Newtonian fluid. Also, the higher the surface potential of a particle, the more significant the non Newtonian nature of a Carreau fluid on its electrophoretic mobility. PMID- 15350057 TI - Preparation of micrometer-sized poly(methyl methacrylate) particles with amphoteric initiator in aqueous media. AB - A previously proposed method based on soap-free emulsion polymerization with an amphoteric initiator for producing micrometer-sized polystyrene particles was extended to application with methyl methacrylate (MMA). The aggregation and dispersion stability of polymer particles, which have ionizable groups arising from initiator radicals, can be controlled by adjusting the pH of the reaction system accompanied with the addition of ionic monomer. Polymerizations were carried out with 2,2'-azobis[N-(2-carboxyethyl)-2-methylpropionamidine] tetrahydrate amphoteric initiator, NH(3)/NH(4)Cl pH buffer, and sodium p styrenesulfonate anionic monomer (NaSS) in ranges of MMA concentration (0.58-2.32 kmol/m(3) H(2)O) and NH(3) concentration (2.5-20 mol/m(3) H(2)O) at fixed concentrations of 5 mol/m(3) H(2)O initiator, 10 mol/m(3) H(2)O NH(4)Cl, and 1 mol/m(3) H(2)O NaSS at 65 degrees C. The addition of NaSS during the polymerization could improve stability in dispersion of particles, which coagulated in the absence of NaSS after the disappearance of monomer drops. An increase in the monomer concentration in the present method could enlarge the particle size without lowering the monodispersity of the particle size distribution. On the other hand, an increase in NH(3) concentration decreased the particle size. The highest monodispersity of particle sizes was obtained at a NH(3) concentration of 5 mol/m(3) H(2)O, which gave an average size of 1.5 microm and a coefficient of variation of particle size distribution of 2.2% that was much smaller than the standard criterion for monodispersity, 10%. PMID- 15350059 TI - A simulation study of electrostatic effects on mixed ionic micelles confined between two parallel charged plates. AB - Confined colloidal systems have been the subject of extensive theoretical and experimental research, and the recent observation of long-range like-charge attraction in such systems has only highlighted their peculiar behavior. On the other hand, surfactant solutions are often used in small confined space, yet their behavior in confinement has received relatively little attention. A distinct feature of confined self-assembling systems is that the aggregates are capable of adjusting their composition, size, and shape in response to their external environment, which may lead to very different phase characteristics compared to bulk solutions. The primary objective of this study is to explore the effects of varying micelle composition on the structural behavior of a confined mixed ionic micellar solution. Mesoscale canonical Monte Carlo simulations were used to probe the structure of the confined solution, while a molecular thermodynamic model was used to systematically account for the change in micelle size as we varied its composition. Significant micelle ordering was found under certain conditions, which implies that large deviations from the minimum-energy micelle configuration may not be entropically favorable. Accumulation of micelles along the midplane was observed when the confining walls are weakly charged, suggesting that micelle shape transformation should be considered in more detail. On the other hand, with high wall charge density, apparent attraction was found between like-charged micelles and wall. These findings point to the need for a more quantitative theoretical treatment in describing surfactant self-assembly in confined geometries. PMID- 15350060 TI - Study of poly(N,N-dimethylacrylamide)/CdS nanocomposite organic/inorganic gels. AB - CdS nanoparticles have been synthesized and stabilized in poly(N,N dimethylacrylamide) hydrogels. The properties of the composite material have been characterized by UV-vis spectroscopy, scanning electron microscopy, X-ray diffraction, thermogravimetric analysis, and steady-state and time-resolved luminescence spectroscopy. This material can be obtained in three different states: swollen, shrunk, and freeze-dried. The swollen and the freeze-dried states correspond to a nanocomposite organic/inorganic (wet or dry) gel containing CdS nanoparticles of approximately 50 nm diameter while the shrunk state is a two-phase system containing CdS crystals, which precipitate forming interesting geometrical shapes. PMID- 15350061 TI - Stabilization of carbon dioxide-in-water emulsions with silica nanoparticles. AB - Stable carbon dioxide-in-water emulsions were formed with silica nanoparticles adsorbed at the interface. The emulsion stability and droplet size were characterized with optical microscopy, turbidimetry, and measurements of creaming rates. The increase in the emulsion stability as the silica particle hydrophilicity was decreased from 100% SiOH to 76% SiOH is described in terms of the contact angles and the resulting energies of attachment for the silica particles at the water-CO(2) interface. The emulsion stability also increased with an increase in the particle concentration, CO(2) density, and shear rate. The dominant destabilization mechanism was creaming, whereas flocculation, coalescence, and Ostwald ripening played only a minor role over the CO(2) densities investigated. The ability to stabilize these emulsions with solid particles at CO(2) densities as low as 0.739 g/mL is particularly relevant in practical applications, given the difficulty in stabilizing these emulsions with surfactants, because of the unusually weak solvation of the surfactant tails by CO(2). PMID- 15350062 TI - Impacting the length of wormlike micelles using mixed surfactant systems. AB - The effect of adding an alcohol ethoxylate nonionic surfactant (C(18)E(18)) to aqueous solutions of a cationic surfactant, erucyl bis(hydroxyethyl) methylammonium chloride (EHAC,CH(3)(CH(2))(7)(CH)(2)(CH(2))(12)N(+) (CH(2)CH(2)OH)(2)CH(3)Cl(-)), was studied using small-angle neutron scattering (SANS), steady-state rheology, and cryo-transmission electron microscopy (Cryo TEM). This cationic surfactant has the ability to self-assemble into giant wormlike micelles in the presence of an electrolyte, such as KCl. In salt-free solutions, the mixture of the two surfactants gave rise to spherical micelles. The scattering curves obtained were fitted with a polydisperse core-shell model combined with a Hayter Penfold potential. The inner and outer radii were found to be dependent on the surfactant ratio. In the presence of KCl, mixed wormlike micelles were formed. However, further addition of C(18)E(18) promoted the breaking of the micellar worms with the appearance of a structure peak in the scattering curves. In addition, it was found that the low shear viscosity is decreased upon addition of the alcohol ethoxylate nonionic surfactant. These findings are in good qualitative agreement with the Cryo-TEM images. The results show that the addition of the nonionic surfactant to the system is a method of controlling the worm length. PMID- 15350063 TI - Organization of four thermotropic liquid crystals of different polarities on model liquid and solid surfaces. AB - The thermodynamic and surface properties of four structurally related thermotropic liquid crystals (LC) were investigated to understand their organization at gas-liquid and gas-solid interfaces. In this study, LC with a benzoyloxy azobenzene mesogenic core substituted with heptyloxy and/or dioxyethylene ether groups were used. The propensity of the LC to form self assembled multilayers was demonstrated in the films spread at the air/aqueous interface using the Langmuir technique and Brewster angle microscopy and on the solid surfaces of Chromosorb WHP and silica, using differential scanning calorimetry. On the basis of the results obtained, a molecular recognition mechanism underlying separation processes using LC as selectors in gas chromatography is proposed. PMID- 15350064 TI - LB films of rodlike phthalocyanine aggregates: specular X-ray reflectivity studies of the effect of interface modification on coherence and microstructure. AB - We present here X-ray specular reflectivity (XRR) characterization of the ordering of Langmuir-Blodgett films of the liquid crystalline phthalocyanine (Pc) 2,3,9,10,16,17,23,34-octakis(2-benzyloxyethoxy)copper(II) phthalocyanine, 1, on Si(100) wafers with a native oxide layer and these same substrates modified with monolayers of varying percentages of methyl- and phenyl-terminated silanes. The central copper atom in these Pc's provides for high contrast in X-ray reflectivity for single-bilayer films of 1. The XRR data are modeled as arising from multiple layers of organic material, above and below the rows of copper atoms in the aligned Pc cores; variations in the total thickness of these films, and the spacing between the rows of copper atoms, are attributed to changes in the interaction with the substrate, and changes in the Pc orientation and side chain packing. The most pronounced effect on these parameters comes from variations in the ratio of the phenyl-silane versus methyl-silane content of the substrate modifiers and annealing of these films past their crystalline (K) --> liquid crystalline (LC) mesophase transition temperature. Transfer of multiple bilayers of this Pc leads to additional changes in the thickness of each layer, eventually forming a hexagonal close-packed array, reminiscent of bulk fibers of this material, as revealed by X-ray diffraction (XRD). PMID- 15350065 TI - Mechanisms for gas adsorption and desorption in silica aerogels: the effect of temperature. AB - We present a theoretical study of the adsorption and desorption mechanisms of fluids in silica aerogels, focusing on the effect of temperature. We adopt a coarse-grained lattice description in which the gel structure is generated by a diffusion-limited cluster-cluster aggregation algorithm and the fluid configurations are computed using local mean-field (i.e., density functional) theory. Our calculations reproduce qualitatively the changes in the shape of the hysteresis loops observed with (4)He in gels of varying porosity. We study in detail the morphology of the condensation and evaporation events that correspond to the irreversible processes (avalanches) which are at the origin of the hysteresis. Depending on porosity and temperature, these avalanches may be localized, involve regions that extend beyond the gel correlation length, or even span the entire sample. This makes difficult the characterization of aerogels based on analyzing sorption isotherms. PMID- 15350066 TI - Multilayered thin films that sustain the release of functional DNA under physiological conditions. AB - The development of thin films and coatings that control the release of DNA from the surfaces of materials could have a significant impact on localized approaches to gene therapy. Here, we report multilayered polyelectrolyte assemblies that sustain the release of functional plasmid DNA from the surfaces of model substrates under physiological conditions. Multilayered assemblies consisting of alternating layers of plasmid DNA encoding for enhanced green fluorescent protein (EGFP) and a synthetic degradable polyamine were deposited on planar silicon and quartz substrates using a layer-by-layer fabrication process. Film growth was monitored by ellipsometry and UV spectrophotometry and correlated linearly with the number of polymer and plasmid layers deposited. In general, the thickness of deposited layers was found to be a function of both the pH and the ionic strength of the polyelectrolyte solutions used. Films up to 100 nm thick were investigated in this study. These assemblies erode gradually upon incubation in phosphate buffered saline at 37 degrees C, as determined by ellipsometry and UV spectrophotometry, and sustain the release of incorporated plasmid into the incubation medium for a period of up to 30 h. Characterization of the released plasmid by agarose gel electrophoresis revealed that the DNA was released in a relaxed, open circular, rather than supercoiled, topology; subsequent cell transfection experiments demonstrated that the released plasmid is transcriptionally viable and promotes the expression of EGFP in the COS-7 cell line. These layered materials could represent an approach to the controlled administration of one or more functional DNA constructs from the surfaces of biomedical materials and devices. PMID- 15350067 TI - X-ray nanoscale profiling of layer-by-layer assembled metal/organophosphonate films. AB - The nanoscale structure of multilayer metal/phosphonate thin films prepared via a layer-by-layer assembly process was studied using specular X-ray reflectivity (XRR), X-ray fluorescence (XRF), and long-period X-ray standing wave (XSW) analysis. After the SiO(2) X-ray mirror surfaces were functionalized with a monolayer film terminated with phosphonate groups, the organic multilayer films were assembled by alternating immersions in (a) aqueous solutions containing Zr(4+), Hf(4+), or Y(3+) cations and then (b) organic solvent solutions of PO(3) R-PO(3), where R was a porphyrin or porphyrin-square spacer molecule. The different heavy metal cations provided X-ray fluorescence marker layers at different heights within the different multilayer assemblies. The XSW measurements used a 22 nm period Si/Mo multilayer mirror. The long-period XSW generated by the zeroth-order (total external reflection) through fourth-order Bragg diffraction conditions made it possible to examine the Fourier transforms of the fluorescent atom distributions over a much larger q(z)() range in reciprocal space than previously achieved. PMID- 15350068 TI - Equilibrium interaction of solid surfaces across a polymer melt. AB - Forces across polymer melts are poorly understood despite their importance for adhesion and fabricating composite materials. Using an atomic force microscope (AFM), this interaction was measured for poly(dimethyl siloxane) (PDMS). The structure of the polymer at the surface changed during the first approximately 10 h. Afterward, short-range attractive forces were observed with short-chain PDMS (M(w) = 4200 g/mol). Using PDMS with a molecular weight (M(w) = 18 000 g/mol) above the entanglement limit, we measured a monotonically decaying repulsive force, which indicates that a quasi-immobilized layer had formed at the solid surface. Due to the small radius of curvature of the tip, forces could be measured in equilibrium. PMID- 15350069 TI - Surface modification of nanoporous alumina surfaces with poly(ethylene glycol). AB - Nanoporous alumina surfaces have a variety of applications in biosensors, biofiltration, and targeted drug delivery. However, the fabrication route to create these nanopores in alumina results in surface defects in the crystal lattice. This results in inherent charge on the porous surface causing biofouling, that is, nonspecific adsorption of biomolecules. Poly(ethylene glycol) (PEG) is known to form biocompatible nonfouling films on silicon surfaces. However, its application to alumina surfaces is very limited and has not been well investigated. In this study, we have covalently attached PEG to nanoporous alumina surfaces to improve their nonfouling properties. A PEG-silane coupling technique was used to modify the surface. Different concentrations of PEG for different immobilization times were used to form PEG films of various grafting densities. X-ray photoelectron spectroscopy (XPS) was used to verify the presence of PEG moieties on the alumina surface. High-resolution C1s spectra show that with an increase in concentration and immobilization time, the grafting density of PEG also increases. Further, a standard overlayer model was used to calculate the thickness of PEG films formed using the XPS intensities of the Al2p peaks. The films formed by this technique are less than 2.5 nm thick, suggesting that such films will not clog the pores which are in the range of 70-80 nm. PMID- 15350070 TI - Photochemical reaction, acidichromism, and supramolecular nanoarchitectures in the Langmuir-Blodgett films of an amphiphilic styrylquinoxaline derivative. AB - An amphiphilic styrylquinoxaline derivative, 3-(4-(hexadecyloxy)styryl)quinoxalin 2(1H)-one (SQC16), was newly synthesized to investigate their photochemical and gas responsive properties in organized molecular films. It was observed that SQC16 can spread as a monolayer on the subphases with various pH values and be subsequently transferred onto solid substrates. While SQC16 showed predominantly reversible trans-cis photoisomerization in methanol solution, it showed both photoisomerization and photodimerization in Langmuir-Blodgett (LB) films. Photodimerization was only observed in the LB film due to the face-to-face arrangement of the functional headgroup in the LB film, and the process was irreversible. In addition, the LB film showed acidichromism, i.e., when the film was exposed to HCl gas its color changed from yellow to red, and the color could be recovered after exposure to NH(3) gas. The process was reversible and could be repeated many times. An interesting surface morphology of the SQC16 LB film was revealed. It was observed that SQC16 can form nanowire architecture in the transferred one-layer LB film. This morphology can be changed upon photoirradiation or in gas reactions. Through the atomic force microscopy measurements it was suggested that the photodimerization predominantly occurred from the nanowire structures, while during the acidichromism the reaction occurred preferentially in the flat region. X-ray diffraction studies revealed that while layer distance showed a slight change for the LB film during acidichromism and photoreaction, the layer structure of SQC16 LB film was retained. PMID- 15350071 TI - Thin film interference of colloidal thin films. AB - A stairlike colloidal crystal thin film composed of poly(styrene-methyl methacrylate-acrylic acid) (P(St-MMA-AA)) monodispersed colloids was fabricated on an inclined silicon substrate. Different bright colors were observed on the various parts of the film with different layers as white light irradiated perpendicularly on it. The relationship between the colors and layers of the film was investigated and discussed according to the principle of thin film interference. On the basis of the phenomenon of thin film interference, a one layer colloidal film having uniform color was researched and it would display diverse colors before and after swollen by styrene (St). A circular stairlike colloidal film was achieved to mimic the colors of the peacock tail feather. PMID- 15350072 TI - Adsorption of aromatic counterions at the surfactant/water interface: a neutron reflectivity study of hydroxybenzoate and chlorobenzoate counterions at the hexadecyl trimethylammonium surfactant/water interface. AB - Specular neutron reflectivity has been used to investigate the adsorption of the aromatic counterions hydroxybenzoate and chlorobenzoate at the hexadecyl trimethylammonium bromide surfactant monolayer/water interface. The degree of counterion binding and the location of the counterions at the interface are shown to depend on the isomeric form of the counterion. For hydroxybenzoate, the para substituted counterion is located within the headgroup region of the surfactant monolayer, and there is of order one counterion for every two surfactant ions. For the ortho-substituted counterion, the degree of counterion binding is higher. There is of order 0.85 counterions for each surfactant ion, and the counterion is located within the hydrophobic region of the monolayer, some 5 A from the center of the headgroup distribution. Similar results were found for the chlorobenzoate counterion, but in that case it was the para-substituted counterion that was more tightly bound and located within the hydrophobic region of the surfactant monolayer. The results for the ortho-substituted hydroxybenzoate and for the para substituted chlorobenzoate are consistent with those previously reported for the para-tosylate. The results are discussed in the context of the ability of the specific aromatic counterion isomer to promote massive micellar growth, and the results shed light on that mechanism. PMID- 15350073 TI - Hydrophilic chain length dependence of the ionic amphiphilic polymer monolayer structure at the air/water interface. AB - Detailed analysis of an interesting poly(methacrylic acid) (PMAA) brush structure in water of a diblock copolymer [(Et(2)SB(m)-b-(MMA)(n), where Et(2)SB is diethylsilacyclobutane] monolayer reported previously was performed by X-ray and neutron reflectometry and indicated that the hydrophilic layer formed with a relatively long PMAA chain is not a simple layer but is divided into two layers, that is, a "carpet"-like dense PMAA layer near the hydrophobic layer and a polyelectrolyte brush layer. The hydrophilic chain length dependence of the diblock copolymer monolayer was analyzed using m:n = 30:x polymer samples, especially of the PMAA double layer structure. With the increase in PMAA chain length in polymer samples, a carpet layer is mainly formed up to n approximately 50. With further increase in the PMAA chain length beyond n approximately 50, a well-defined brush layer appears. On the other hand, the variation in hydrophobic layer thickness with methacrylic acid unit is minimum at the critical PMAA length, that is, n approximately 50 under a constant surface pressure condition. It is strongly suggested that the two discrete layers contribute differently to surface pressure. Furthermore, from the comprehensive viewpoint, the major factor for brush formation was clarified not to be the absolute length of the PMAA chain, but the ratio of PEt(2)SB and PMAA chain length is an important factor. PMID- 15350074 TI - CH3O decomposition on PdZn(111), Pd(111), and Cu(111). A theoretical study. AB - Methanol steam re-forming, catalyzed by Pd/ZnO, is a potential hydrogen source for fuel cells, in particular in pollution-free vehicles. To contribute to the understanding of pertinent reaction mechanisms, density functional slab model studies on two competing decomposition pathways of adsorbed methoxide (CH(3)O) have been carried out, namely, dehydrogenation to formaldehyde and C-O bond breaking to methyl. For the (111) surfaces of Pd, Cu, and 1:1 Pd-Zn alloy, adsorption complexes of various reactants, intermediates, transition states, and products relevant for the decomposition processes were computationally characterized. On the surface of Pd-Zn alloy, H and all studied C-bound species were found to prefer sites with a majority of Pd atoms, whereas O-bound congeners tend to be located on sites with a majority of Zn atoms. Compared to Pd(111), the adsorption energy of O-bound species was calculated to be larger on PdZn(111), whereas C-bound moieties were less strongly adsorbed. C-H scission of CH(3)O on various substrates under study was demonstrated to proceed easier than C-O bond breaking. The energy barrier for the dehydrogenation of CH(3)O on PdZn(111) (113 kJ mol(-)(1)) and Cu(111) (112 kJ mol(-)(1)) is about 4 times as high as that on Pd(111), due to the fact that CH(3)O interacts more weakly with Pd than with PdZn and Cu surfaces. Calculated results showed that the decomposition of methoxide to formaldehyde is thermodynamically favored on Pd(111), but it is an endothermic process on PdZn(111) and Cu(111) surfaces. PMID- 15350075 TI - Two-dimensional hexagonally oriented CdCl2.H2O nanorod assembly: formation and replication. AB - This paper reports a simple bottom-up method that can controllably fabricate 2D hexagonally oriented and randomly distributed CdCl(2).H(2)O nanorods on mica surfaces. The as-formed nanorod assemblies have been successfully replicated into various matrixes, including gold, poly(dimethylsiloxane), and polyurethane. Thus, this method is compatible with soft-lithography towards further applications. PMID- 15350076 TI - High-pressure adsorption of CO2 on NaY zeolite and model prediction of adsorption isotherms. AB - Supercritical carbon dioxide is an efficient solvent for adsorptive separations because it can potentially be used as both the carrier solvent for adsorption and the desorbent for regeneration. Recent results have demonstrated an anomalous peak or "hump" in the adsorption isotherm near the bulk critical point when the adsorption isotherm is plotted as a function of bulk density. This work presents new data for the adsorption and desorption of carbon dioxide in the near-critical region on a crystalline, well-structured adsorbent (NaY zeolite). The results indicate a strong affinity for CO(2) as well as a significant hump near the critical point. The lattice model previously developed by Aranovich and Donohue is applied to analyze the adsorption. PMID- 15350077 TI - DNA-directed protein immobilization on mixed self-assembled monolayers via a streptavidin bridge. AB - The simultaneous detection of multiple analytes is an important consideration for the advancement of biosensor technology. Currently, few sensor systems possess the capability to accurately and precisely detect multiple antigens. This work presents a simple approach for the functionalization of sensor surfaces suitable for multichannel detection. This approach utilizes self-assembled monolayer (SAM) chemistry to create a nonfouling, functional sensor platform based on biotinylated single-stranded DNA immobilized via a streptavidin bridge to a mixed SAM of biotinylated alkanethiol and oligo(ethylene glycol). Nonspecific binding is minimized with the nonfouling background of the sensor surface. A usable protein chip is generated by applying protein-DNA conjugates which are directed to specific sites on the sensor chip surface by utilizing the specificity of DNA hybridization. The described platform is demonstrated in a custom-built surface plasmon resonance biosensor. The detection capabilities of a sensor using this protein array have been characterized using human chorionic gonadotropin (hCG). The platform shows a higher sensitivity in detection of hCG than that observed using biotinylated antibodies. Results also show excellent specificity in protein immobilization to the proper locations in the array. The vast number of possible DNA sequences combine with the selectivity of base-pairing makes this platform an excellent candidate for a sensor capable of multichannel protein detection. PMID- 15350078 TI - Influence of hyperbranched polyesters on the surface tension of polyols. AB - The influence of hyperbranched polyesters with different functional end groups on the surface tension of mixtures with an oligo(ester diol) was investigated. The temperature dependence of the surface tension of the pure components and of the mixtures was measured by a modified Wilhelmy balance technique. The results indicate that the surface tension of the pure hyperbranched polyesters strongly depends on the functionality of the end groups. The functionalization of the hydroxyl end groups by short alkyl chains (methyl, tert-butyl) reduced the surface tension depending on the degree of substitution. The surface tension of the mixtures with the hydroxyl-terminated hyperbranched polyester was slightly increased at higher concentrations of the hyperbranched polymer compared to the surface tension of the pure ester diol. On the other hand, the surface tension of mixtures could be considerably decreased using 1% of hyperbranched polyester polyols partially substituted with short alkyl chains. In that case, the modified hyperbranched polyesters act as surface active agents. On the molecular level, the enrichment of the modified hyperbranched polyester in the surface region was proven by X-ray photoelectron spectroscopy measurements. PMID- 15350079 TI - Counterions in poly(allylamine hydrochloride) and poly(styrene sulfonate) layer by-layer films. AB - The amount of counterions in layer-by-layer (LBL) films of poly(allylamine hydrochloride) (PAH) and poly(styrene sulfonate) (PSS) has been determined with X ray photoelectron spectroscopy (XPS) for films prepared from solutions with various NaCl concentrations. Sodium and chloride counterions are present in LBL films produced from salt solutions, which are located at the surface and in the bulk of the films. The percentage of bulk counterions increases with the ionic strength of the polyelectrolyte before reaching a constant value. The bulk sodium/sulfur percentage ratios tend to 0.8 for samples washed with pure water and for samples washed with NaCl aqueous solutions, while the bulk chlorine/nitrogen percentage ratios tend to 0.5 for the same samples. The ratio between the percentages of polyelectrolyte ionic groups lies close to unity for all samples, indicating that counterions do not contribute to charge compensation in the polyelectrolyte during the adsorption process. The presence of counterions in LBL films is explained by Manning condensation near the polyelectrolyte ionic groups, leading to inter-polyelectrolyte ionic bondings via ionic networks. It is believed that condensation leads to the formation of NaCl crystallites in these LBL films, which was confirmed by X-ray diffraction measurements. PMID- 15350080 TI - Interfacial tension of a nematic liquid crystal/water interface with homeotropic surface alignment. AB - Pendant drop experimental results are presented for the temperature dependence of the interfacial tension between water and the immiscible nematic liquid crystal 4'-pentyl-4-biphenylcarbonitrile (5CB) in the presence of the adsorbed surfactant cetyltrimethylammonium bromide (CTAB). Adsorption of the surfactant lowers the interfacial tension value and is also known from earlier work to induce a transition in liquid crystal surface alignment from planar to homeotropic [Brake et al. Langmuir 2003, 19, 6436.]. Discrepancies exist in the literature regarding the density of 5CB, and the density difference between 5CB and water in any case is very small. However, from the ability to form pendant 5CB drops, one may infer that the density of 5CB exceeds that of water over the entire temperature range studied (28-41 degrees C), in disagreement with the predictions of one earlier report on 5CB. The interfacial tension is shown to exhibit a relative maximum near the bulk 5CB nematic-isotropic transition temperature T(NI), regardless of which published data set of 5CB density values is used to analyze the measurements, with a possible discontinuity in tension occurring at T(NI). The anomalous shape of the interfacial tension curve, depending on the choice of the 5CB density data set, may be quite similar to that recently reported for the interface between 5CB and a hydrophobic, isotropic molten polymer (Rai et al. Langmuir 2003, 19, 7370). PMID- 15350081 TI - Adsorption and desorption of polymer/surfactant mixtures at solid-liquid interfaces: substitution experiments. AB - The adsorption of mixtures of aqueous solutions of cationic hydroxyethylcellulose polymer JR400 and anionic surfactant, sodium dodecyl sulfate, using atomic force microscopy (AFM) has been studied. Samples with various compositions from different regions of the ternary phase diagram presented in our previous work were imaged by atomic force microscopy on freshly cleaved mica, and hydrophobically modified mica and silica in soft-contact mode. A series of "washing" (subsequent injection of compositions with gradually decreasing polymer/surfactant ratio) and "scratching" (mechanical agitation of the surface material with an AFM tip) experiments were performed. It was revealed that the morphology of the adsorbed layer altered in a manner following the changes in morphology in the bulk solution. These changes were evidenced in cluster formation in the layer. The results suggest that the influence of the surface was limited to the formation of the adsorbed layer where the local concentrations of polymer and surfactant were higher than those in the bulk. All further modifications were driven by changes in the mixture composition in bulk. Force measurements upon retraction reveal the formation of network structures within the surface aggregates that will greatly slow structural reequilibration. PMID- 15350082 TI - Phase state effect on adhesion behavior of self-assembled monolayers. AB - The effect of phase state of self-assembled monolayers (SAMs) on adhesion behavior was studied using a combination of atomic force microscopy (AFM) and Johnson-Kendall-Roberts (JKR) methods. The phase state of SAMs was controlled by adjusting the reaction temperature. Order-to-disorder structural transitions in monolayers of n-alkyltrichlorosilanes resulted in dramatic increases in adhesion force and adhesion hysteresis, which represents the first report of alterations in adhesion properties due to phase changes of monolayers without any effect of chain length and surface heterogeneity. This increase in mechanical deformation of the disordered monolayer is understood to be caused by increases in (1) molecular contact between the AFM tip and a disordered monolayer due to the more deformable state of the latter and (2) monolayer deformation during unloading by the JKR probe lens. Adhesion hysteresis was found to have greater sensitivity toward the unloading rate for disordered monolayers. The occurrence of maximum hysteresis at faster rates proves that monolayer chain mobility increases with structural disorder, resulting in increased mechanical deformation. PMID- 15350083 TI - Simultaneous tailoring of surface topography and chemical structure for controlled wettability. AB - Wettability was controlled in a rational manner by individually and simultaneously manipulating surface topography and surface chemical structure. The first stage of this research involved the adsorption of charged submicrometer polystyrene latex particles to oppositely charged poly(ethylene terephthalate) (PET) film samples to form surfaces with different topographies/roughness; adsorption time, solution pH, solution ionic strength, latex particle size, and substrate charge density are external variables that were controlled. The introduction of discrete functional groups to smooth and rough surfaces through organic transformations was carried out in the second stage. Amine groups ( NH(2)) and alcohol groups (-OH) were introduced onto smooth PET surfaces by amidation with poly(allylamine) and adsorption with poly(vinyl alcohol) (PVOH), respectively. On latex particle adsorbed surfaces, a thin layer of gold was evaporated first to prevent particle redistribution before chemical transformation. Reactions with functionalized thiols and adsorption with PVOH on patterned gold surfaces successfully enhanced surface hydrophobicity and hydrophilicity. Particle size and biomodal particle size distribution affect both hydrophobicity and hydrophilicity. A very hydrophobic surface exhibiting water contact angles of 150 degrees /126 degrees (theta(A)/theta(R)) prepared by adsorption of 1-octadecanethiol and a hydrophilic surface with water contact angles of 18 degrees /8 degrees (theta(A)/theta(R)) prepared by adsorption of PVOH were prepared on gold-coated surfaces containing both 0.35 and 0.1 microm latex particles. The combination of surface topography and surface-chemical functionality permits wettability control over a wide range. PMID- 15350084 TI - Lattice boltzmann study on the contact angle and contact line dynamics of liquid vapor interfaces. AB - The moving contact line problem of liquid-vapor interfaces was studied using a mean-field free-energy lattice Boltzmann method recently proposed [Phys. Rev. E 2004, 69, 032602]. We have examined the static and dynamic interfacial behaviors by means of the bubble and capillary wave tests and found that both the Laplace equation of capillarity and the dispersion relation were satisfied. Dynamic contact angles followed the general trend of contact line velocity observed experimentally and can be described by Blake's theory. The velocity fields near the interface were also obtained and are in good agreement with fluid mechanics and molecular dynamics studies. Our simulations demonstrated that incorporating interfacial effects into the lattice Boltzmann model can be a valuable and powerful alternative in interfacial studies. PMID- 15350085 TI - Cadmium ion adsorption on different carbon adsorbents from aqueous solutions. Effect of surface chemistry, pore texture, ionic strength, and dissolved natural organic matter. AB - Adsorption of Cd(II) species at pH = 5 was studied on three carbon adsorbents: granular activated carbon, activated carbon fiber, and activated carbon cloth. As received and oxidized adsorbents were used. Cd(II) adsorption greatly increased after oxidation due to the introduction of carboxyl groups. The use of a buffer solution to control the pH introduced some changes in the surface chemistry of carbons through the adsorption of one of the compounds used, biphthalate anions. The increase in ionic strength reduced Cd(II) uptake on both as-received and oxidized carbons due to a screening of the electrostatic attractions between the Cd(II) positive species and the negative surface charge, which in the case of as received carbons derived from the biphthalate anions adsorbed and in the oxidized ones from the carboxyl groups. Tannic acid was used as a model compound for natural organic matter. Its adsorption was greatly reduced after oxidation, and most of the carbon adsorbents preadsorbed with tannic acid showed an increase in Cd(II) uptake. In the case of competitive adsorption between Cd(II) species and tannic acid molecules, there was a decrease in Cd(II) uptake on the as-received carbon whereas the contrary occurred with the oxidized carbons. These results illustrate the great importance of carbon surface chemistry in this competitive adsorption process. Finally, under all experimental conditions used, when the adsorption capacity of carbons was compared under the same conditions it increased in the following order: granular activated carbon < activated carbon fiber < activated carbon cloth. PMID- 15350086 TI - Photoinduced phase separation and miscibility in the condensed phase of a mixed Langmuir monolayer. AB - We report our studies on the mixed Langmuir monolayer of mesogenic molecules, p (ethoxy)-p-phenylazo phenyl hexanoate (EPPH) and octyl cyano biphenyl (8CB), employing the techniques of surface manometry and Brewster angle microscopy. Our studies show that the mixed monolayer exhibits higher collapse pressures for certain mole fractions of EPPH in 8CB as compared to individual monolayers. Also, a considerable reduction in the area per molecule is seen in the mixed monolayer, indicating a condensed phase. We have also studied the photostability of the mixed monolayer at different initial surface pressures. The mixed monolayer, under alternate cycles of UV and visible illumination, exhibits changes in surface pressures. This is due to the photoinduced transformation of EPPH isomers in the mixed monolayer. Our in-situ Brewster angle microscope studies for 0.5 mole fraction of EPPH in 8CB show a phase separation in the UV and a miscible phase in the visible, at low surface pressures ( approximately 5 mN/m). At higher surface pressures ( approximately 10 mN/m), under UV illumination, we find a phase separation which does not revert to a miscible phase under visible illumination. PMID- 15350087 TI - On the monomer density of grafted polyelectrolyte brushes and their interactions. AB - Most of the modern theories of grafted polyelectrolyte brushes are valid only for moderate stretching of the polyelectrolyte. However, particularly at low ionic strength and high grafting densities, even a moderate charge of the polyelectrolyte can generate a strong stretching. A simple mean field model for strongly stretched grafted polyelectrolyte brushes is suggested, based on an approximate calculation of the partition function of a polyelectrolyte chain. It is shown that the average Boltzmann factor of a possible chain configuration can be approximated by the Boltzmann factor of a configuration with a constant monomer distribution, for which the free energy can be readily obtained. The monomer density in the brush and the interaction between two surfaces with grafted polyelectrolyte brushes could be calculated as a statistical average over all possible configurations. Some simple analytical results are derived, and their accuracy is examined. The dependence of the brush thickness on the electrolyte concentration is investigated, and it is shown that the trapping of a fraction of counterions in the brush influences strongly the thickness of the brush. When two surfaces with grafted polyelectrolyte brushes approach each other more rapidly than the ion diffusion parallel to the surface, the trapping of the counterions between the brushes can affect the interactions by orders of magnitude. PMID- 15350088 TI - Organization and orientation of amphiphilic push-pull chromophores deposited in Langmuir-Blodgett monolayers studied by second harmonic generation and atomic force microscopy. AB - Orientation and organization of two amphiphilic push-pull chromophores mixed with two phospholipids (dipalmitoylphosphatidylcholine and dioleoylphosphatidylcholine) in Langmuir-Blodgett (LB) monolayers are investigated by second harmonic generation. The LB monolayers have also been characterized by atomic force microscopy and UV-vis spectroscopy. The effective molecular orientations and hyperpolarizabilities of the chromophores are studied as a function of the phospholipid concentrations. The experimental results are discussed within the frame of a model of orientational distribution of the chromophores which gives the orientational mean angle and bounds on the orientational disorder. The mean orientation of the chromophores is found to be within 45-55 degrees whereas their hyperpolarizability coefficients, measured with respect to quartz, are estimated to be in the range (0.3-0.7) x 10(-27) esu taking account of the maximal orientational disorder. PMID- 15350089 TI - The nature of alkanethiol self-assembled monolayer adsorption on sputtered gold substrates. AB - A detailed study of the self-assembly and coverage by 1-nonanethiol of sputtered Au surfaces using molecular resolution atomic force microscopy (AFM) and scanning tunneling microscopy (STM) is presented. The monolayer self-assembles on a smooth Au surface composed predominantly of [111] oriented grains. The domains of the alkanethiol monolayer are observed with sizes typically of 5-25 nm, and multiple molecular domains can exist within one Au grain. STM imaging shows that the (4 x 2) superlattice structure is observed as a (3 x 2) structure when imaged under noncontact AFM conditions. The 1-nonanethiol molecules reside in the threefold hollow sites of the Au[111] lattice and aligned along its [112] lattice vectors. The self-assembled monolayer (SAM) contains many nonuniformities such as pinholes, domain boundaries, and monatomic depressions which are present in the Au surface prior to SAM adsorption. The detailed observations demonstrate limitations to the application of 1-nonanethiol as a resist in atomic nanolithography experiments to feature sizes of approximately 20 nm. PMID- 15350090 TI - Complex formation between dodecylpyridinium chloride and multicharged anionic planar substances. AB - The complex formation between dodecylpyridinium chloride (DPC) and multicharged anionic planar substances, 14 azo dyes and 3 benzene- or naphthalenesulfonates, has been studied by the potentiometric titration using a surfactant selective electrode. The agreement between the observed maximum binding number and the number of anionic charges (n) on dye molecules showed n:1 complex formation. The binding isotherms were found to be composed of two types of binding; one is the noncooperative binding observed at low surfactant concentrations and the other is the cooperative binding at the higher concentrations. The microscopic binding constant for the noncooperative binding was found to take the values in the range of 50-200 mol(-)(1) dm(3) for many of the substances, but, takes more large values up to 2500 mol(-)(1) dm(3) for the substances which have a large hydrophobic part or the structure of separate hydrophobic and hydrophilic regions. A multiple regression analysis showed that the data of the corresponding standard free energy change of binding were well interpreted by the equation (in unit of kJ mol(-)(1)) DeltaG degrees = - 5.85 log P(S) - 1.68 log P(D) - 2.12z + 28.4, where P(S) and P(D) are the partition coefficients of the surfactants and planar substances in the 1-octanol/water system and z is the number of anionic charges on the planar molecules. At the beginning of the cooperative binding, precipitate formation was observed for almost all of the present systems. Among these, some of the dyes having the structure of separate hydrophobic and hydrophilic regions formed a needlelike crystal, which was accompanied by a hysteresis phenomenon in the binding isotherm. The stable complex formation by both the hydrophobic and electrostatic interactions between the surfactant and the planar substances was found to be important for the crystal formation. Depending on the manner of arrangement of the charged groups on the planar substances, the origin of the binding cooperativity was ascribed to the interactions between surfactants bound to one planar-substance molecule or to the association of the complexes. It was also found that the present small binding systems are useful as the model of ligand binding to protein local structures. PMID- 15350091 TI - Pressure versus length isotherms of homogenous and mixed one-dimensional dipolar monolayers. AB - We demonstrate a novel method for compressing and expanding microscopic one dimensional monolayers consisting of a finite number of aligned magnetic dipoles using a pair of microscopic magnetic barriers. By measuring the interaction between the beads and the barriers, we are able to determine the pressure of the dipolar monolayers. Our sensor can measure one-dimensional pressure in the femto and piconewton regime and is used to probe both homogeneous and mixed monolayers consisting of magnetic beads with diameters 1.0 microm and 2.8 microm. The larger beads appear to be well-described by a formalism taking into account magnetic dipolar interactions, whereas for smaller beads, such a simple picture does not hold. Upon compressing the monolayer above a certain density, it forms a bilayer. This process is governed by steric interactions or dipolar interactions, depending on the applied magnetic field. We also found odd-even effects, where the number of beads in the monolayer determines the initial structure of the bilayer. PMID- 15350092 TI - Branched fluoropolymer-Si hybrids via surface-initiated ATRP of pentafluorostyrene on hydrogen-terminated Si(100) surfaces. AB - Linear, branched, and arborescent fluoropolymer-Si hybrids were prepared via surface-initiated atom transfer radical polymerization (ATRP) from the 4 vinylbenzyl chloride (VBC) inimer and ClSO(3)H-modified VBC that were immobilized on hydrogen-terminated Si(100), or Si-H, surfaces. The simple approach of UV induced coupling of VBC with the Si-H surface provided a stable, Si-C bonded monolayer of "monofunctional" ATRP initiators (the Si-VBC surface). The aromatic rings of the Si-VBC surface were then sulfonated by ClSO(3)H to introduce sulfonyl chloride (-SO(2)Cl) groups and to give rise to a monolayer of "bifunctional" ATRP initiators. Kinetics study indicated that the chain growth of poly(pentafluorostyrene) from the functionalized silicon surfaces was consistent with a "controlled" or "living" process. The chemical composition and functionality of the silicon surface were tailored by the well-defined linear and branched fluoropolymer brushes. Atomic force microscopy images revealed that the surface-initiated ATRP of pentafluorostyrene (PFS) had proceeded uniformly on the Si-VBC surface to give rise to a dense and molecularly flat surface coverage of the linear brushes. The uniformity of surfaces with branched brushes was controlled by varying the feed ratio of the monomer and inimer (VBC in the present case). The living chain ends on the functionalized silicon surfaces were used as the macroinitiators for the synthesis of diblock copolymer brushes, consisting of the PFS and methyl methacrylate polymer blocks. PMID- 15350093 TI - Mimicking the lotus effect: influence of double roughness structures and slender pillars. AB - Surface roughness is known to amplify hydrophobicity. The apparent contact angle of a drop on a rough surface is often modeled using either Wenzel's or Cassie's formulas. These formulas, along with an appropriate energy analysis, are critical in designing superhydrophobic substrates for applications in microscale devices. In this paper we propose that double (or multiple) roughness structures or slender pillars are appropriate surface geometries to develop "self-cleaning" surfaces. The key motivation behind the double structured roughness is to mimic the microstructure of superhydrophobic leaves (such as lotus). Theoretical analysis similar to that presented in the paper can be used to obtain optimal geometric parameters for the rough surface. The calculation procedure should result in surface geometries with excellent water repellent properties. PMID- 15350094 TI - Exceptional gas permeation selectivity of a glued Langmuir-Blodgett bilayer by pH control. AB - The monolayer properties of 5,11,17,23,29,35-hexakis[(N,N,N-trimethylamonium)-N methyl-37,38,39,40,41,42-hexakis-n-hexadecyloxy-calix[6]arene hexachloride (1) have been characterized over aqueous solutions of poly(acrylic acid) (PAA) and poly(methacrylic acid) (PMAA) as a function of pH. At high pH values (e.g., pH 10), such monolayers show relatively low surface viscosities. At low pH (e.g., pH 4.4), these monolayers exhibit relatively high surface viscosities. The barrier properties of single Langmuir-Blodgett bilayers of 1, which have been ionically cross-linked (i.e., "glued together") with PAA were found to correlate with changes in surface viscosity. Thus, bilayers that were fabricated under low pH conditions exhibited high permeation barriers and high permeation selectivity with respect to He and N(2). Given the extreme thinness of these glued bilayers (ca. 6 nm), the optimized He/N(2) selectivity of ca. 1000 is extraordinary. These results, taken together, demonstrate the feasibility of fine tuning the surface viscosity of monolayers of 1, and also the barrier properties of corresponding glued bilayers, by adjusting the pH of an aqueous subphase that contains a weak polyacid. PMID- 15350095 TI - Real-time observation of lamellar branching induced by an AFM tip and the stability of induced nuclei. AB - Branching of edge-on lamellae of poly(bisphenol A octane ether) (BA-C8) was studied at room temperature (21 +/- 1 degrees C) using real-time tapping mode atomic force microscopy (AFM). Lamellar branches were shown to develop from induced nuclei that had originated from lamellar defects, namely protruding cilia and loose loops, of parent lamellae. Induced nuclei appeared and also disappeared a phenomenon observed first time by AFM. Induced nuclei are one of the possible origins for the branching of lamellae. Branching of edge-on lamellae can be induced by an AFM tip though the adjustment of the set-point amplitude ratio (r(sp)). Using an r(sp) smaller than 0.65, defects could be created on the lamellar surface, which gave rise to lamellar branches. PMID- 15350096 TI - Surface properties of hydrous manganite (gamma-MnOOH). A potentiometric, electroacoustic, and X-ray photoelectron spectroscopy study. AB - The acid-base characteristics of the manganite (gamma-MnOOH) surface have been studied at pH above 6, where dissolution is negligible. Synthetic microcrystalline particles of manganite were used in the experiments. From potentiometric titrations, electrophoretic mobility measurements, and X-ray photoelectron spectroscopy (XPS), a one pK(a) model was constructed that describes the observed behavior. The data show no ionic strength effect at pH < 8.2, which is the pH at the isoelectric point (pH(iep)), but ionic strength effects were visible above this pH. To explain these observations, Na(+) ions were suggested to form a surface complex. The following equilibria were established: =MnOH(2)(+1/2) right harpoon over left harpoon =MnOH(-)(1/2) + H(+), log beta(0) (intr.) = -8.20; =MnOH(2)(+1/2) + Na(+) right harpoon over left harpoon =MnOHNa(+1/2) + H(+), log beta(0) (intr.) = -9.64. The excess of Na(+) at the surface was supported by XPS measurements of manganite suspensions containing 10 mM NaCl. The dielectric constant of synthetic manganite powder was also determined in this study. PMID- 15350097 TI - Surface characterization, modification chemistry, and separation performance of polyimide and polyamidoamine dendrimer composite films. AB - 6FDA-polyimide films modified by polyamidoamine (PAMAM) dendrimers with generations of 0, 1, and 2 are reported in this article. The actual molecular conformation and bulk size of these three generation dendrimers immobilized on polyimide surface were characterized by atomic force microscopy. After comparing with the results of dynamic simulation, we believe that the disk-shape cluster structure of dendrimers has been developed on the polymer surfaces. The amidation and cross-linking reaction between dendrimers and polyimide were examined and quantified by X-ray photoelectron spectroscopy, attenuated total reflection Fourier transform infrared spectroscopy, and gel content measurements. Modification time and the generations of PAMAM dendrimer have been verified as two important factors in determining the properties of modified polyimide films. These modified polyimide films exhibit excellent gas separation performance. The ideal selectivity of He/N(2) increases tremendously to about 200% as compared to that of the original polyimide film. Particularly, the separation performance of CO(2)/CH(4) gas pair can be improved beyond the upper bond limit possibly due to the strong interactions of dendrimer molecules with CO(2), which was verified by sorption tests. PMID- 15350098 TI - Functionalized multiwall carbon nanotube/gold nanoparticle composites. AB - Multiwall carbon nanotubes (MWCNTs) were chemically oxidized in a mixture of sulfuric acid and nitric acid (3:1) while being ultrasonicated. The effect of oxidative ultrasonication at room temperature on development of functional groups on the carbon nanotubes was investigated. The dispersability and the carboxylic acid group concentration of functionalized MWCNTs (fMWNTs) varied with reaction time. The concentration of carboxylic acid groups on fMWNTs increased from 4 x 10(-4) mol/g of fMWNTs to 1.1 x 10(-3) mol/g by doubling the treatment period from 4 to 8 h. The colloidal stability of aqueous fMWCNTs dispersions was enhanced through elongated oxidation. fMWCNTs that were reacted longer than 4 h did not precipitate in aqueous media for at least 24 h. The layer-by-layer self assembly of polyelectrolytes on fMWCNTs was characterized by zeta potential measurements. The zeta potential of fMWCNTs changed from negative charge to positive charge when cationic polyelectrolytes were self-assembled on their surface. With addition of anionic polyelectrolytes, cationic polyelectrolyte coated fMWCNTs showed the expected charge reversal as expected for multilayer self-assembly. Complex formation of positively charged gold nanoparticles and negatively charged fMWCNTs was achieved with and without polyelectrolyte coatings by electrostatic interaction. The complex formation was characterized by high resolution transmission electron microscopy and energy-dispersive X-ray spectroscopy. The here found complex formation of positively charged colloidal gold and defect sites on fMWNTs indicates the location of functional groups on carbon nanotubes. It is suggested that positively charged colloids such as gold nanoparticles could be used for detection of defect sites on carbon nanotubes. PMID- 15350099 TI - Electrochemical oxidation of aliphatic amines and their attachment to carbon and metal surfaces. AB - The electrochemical oxidation of aliphatic amines (primary, secondary, and tertiary) has been investigated by cyclic voltammetry and preparative electrolysis. The oxidation mechanisms have been established, and the lifetimes of the radical cations have been measured for secondary and tertiary amines. These results have been put in parallel with the attachment of amines to glassy carbon, Au, and Pt electrodes by cyclic voltammetry, X-ray photoelectron spectroscopy (XPS), and infrared reflection-absorption spectroscopy (IRRAS). It is then possible to show that it is not the radical cation but the radical obtained after the deprotonation which reacts with the electrode surface. XPS results also point to the existence of a covalent bond between Au or Pt and the organic moiety. PMID- 15350100 TI - Identification of surface states on finely divided supported palladium catalysts by means of inelastic incoherent neutron scattering. AB - The purpose of the present investigation was to utilize the inelastic incoherent neutron scattering (INS) technique to reveal changes at the surface of technical catalysts under the influence of hydrogen in gas/solid interactions and during chemical reactions in a liquid-phase process. The formation and the properties of supported palladium hydride and changes of the hydrogen-related surface chemistry of the corresponding activated carbon supports in 20% Pd/C catalysts after short term and long-term hydrogen cycling at different hydrogen pressures and temperatures were studied. The spectra indicate that hydrogenation of the activated carbon support by hydrogen spillover occurs to, partly, give a material that strongly resembles a-C:H (amorphous hydrogenated carbon). Indications for different relaxation phenomena and long-range phase coherence inside of supported particles of palladium hydride compared to hydrogenated palladium black were obtained. A 5% Pd/C catalyst after use in C-C coupling reactions, the Heck reaction of bromobenzene and styrene to stilbenes, was also studied after subsequent solvent extraction. Evidence for a preferential adsorption and accumulation of cis-stilbene at the catalyst surface was obtained. INS allows identification of a certain isomer from a complex reaction mixture preferentially adsorbed at the surface of a finely divided industrial heterogeneous catalyst. PMID- 15350101 TI - Formation of supramolecular cavitands on copper electrode surfaces. AB - The adsorption of 1,1'-dibenzyl-4,4'-bipyridinium molecules (dibenzyl-viologen or DBV(2+) for the sake of simplicity) on chloride precovered Cu(100) has been studied in an electrochemical environment by means of cyclic voltammetry and in situ scanning tunneling microscopy. DBV(2+) spontaneously forms a highly ordered phase on the chloride c(2 x 2) adlayer at potentials close to the onset of the copper dissolution reaction when the pure supporting electrolyte (10 mM HCl/5 mM KCl) is exchanged by one also containing DBV(2+). This ordered phase can be described by a ( radical 53 x radical 53)R15.9 degrees unit cell relating the organic adlayer to the chloride c(2 x 2) structure underneath or alternatively by a ( radical 106 x radical 106)R29.05 degrees unit cell relating the organic layer to the Cu(1 x 1) substrate structure. Thus, the negatively charged chloride layer acts as a template for the adsorption and phase formation of DBV(2+). Compared to the copper-chloride interaction, the DBV(2+)-chloride interaction appears to be weaker since the organic layer can be easily removed from the surface by the tunneling tip when drastic tunneling conditions (low bias voltage, high tunneling current) are applied. A key structural element of the DBV(2+) adlayer is an assembly of four individual DBV(2+) molecules forming square-shaped supramolecular units with pronounced cavities in their center. Characteristically, the supramolecular assemblies reveal a preferential rotational orientation resulting in the appearance of two chiral forms of these assemblies. Furthermore, these two chiral supramolecular assemblies occur in two mirrored domains of the ( radical 53 x radical 53)R15.9 degrees structure. It can be assumed that these viologen-based supramolecular architectures can be used as potential host cavitands for the inclusion of smaller organic molecules. PMID- 15350102 TI - Electrochemical dimerization of 2-(2'-thienyl)pyridine adsorbed on Au(111) observed by in situ fluorescence. AB - The study of heterodentate molecules adsorbed on metal electrodes provides an opportunity to expand the functionality of modified surfaces while offering insights into the surface and intramolecular electronic interactions of organic adsorbates. The adsorption of 2-(2'-thienyl)pyridine, a molecule containing both pyridine and thiophene moieties, on a Au(111) electrode is reported. Adsorption was characterized by electrochemistry in neutral and basic aqueous electrolyte and was compared to that of pyridine. The aqueous electrochemistry of thiophene on Au(111) was also characterized for comparison purposes. At negative potentials, in the presence of 2-(2'-thienyl)pyridine, a diffuse, pi-bonded monolayer was formed, and a phase transition to a close-packed N- and/or S-bonded configuration was observed near -0.4 V in a 1 mM solution of adsorbate, similar to that seen in pyridine on Au(111). The thiophene-like oxidative dimerization of the molecule was confirmed at positive potentials using in situ fluorescence microscopy by comparison with the spectrum of the chemically synthesized dimer. PMID- 15350103 TI - Measurement of cell migration on surface-bound fibronectin gradients. AB - A novel technique for the quantitative observation of cell migration along linear gradient substrates functionalized with adhesive proteins is presented. Gradients of the cell adhesion molecule fibronectin are generated by the cross diffusion of functionalizable alkanethiols on gold and characterized by X-ray photoelectron spectroscopy and surface plasmon resonance. Two distinct migration assays are described that characterize the movement of either sparsely populated noncontacting cells or a confluent monolayer of cells into free space. The drift speed of bovine aortic endothelial cells is measured and shown to increase along a fibronectin gradient when compared to a uniform control substrate using both assays. The results of these experiments establish reproducible conditions for studies of cell migration on gradients of surface-bound ligands. PMID- 15350104 TI - Brewster angle microscopy of calcium oxalate monohydrate precipitation at phospholipid monolayer phase boundaries. AB - The precipitation of calcium oxalate monohydrate (COM) at phospholipid monolayers confined to the air/water interface is observed in situ with the aid of Brewster angle microscopy. COM crystals appear as bright objects that are easily identified and quantified to assess the effects of different conditions on crystallization. Crystal precipitation was monitored at monolayers of 1,2 dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in liquid condensed (LC) and liquid expanded (LE) phases. Within the LC phase, higher pressures reduce the incidence of crystallization at the interface, implying that within this phase precipitation is enhanced by higher compressibility or fluidity of the monolayer. Precipitation at biphasic LC/LE and LE/gas (G) monolayers was also studied. COM appears preferentially at phase boundaries of the DPPC LC/LE and LE/G monolayers. However, when an LC/LE phase boundary is created by two different phospholipids that are phase segregated, such as DPPC and 1,2-dimyristoyl-sn-glycero-3 phosphocholine, crystal formation occurs away from the interface within the DPPC LC phase. It is suggested that COM growth at phase boundaries is preferred only when there is molecular exchange between the phases. PMID- 15350105 TI - Controlled grafting of well-defined epoxide polymers on hydrogen-terminated silicon substrates by surface-initiated ATRP at ambient temperature. AB - Controlled grafting of well-defined epoxide polymer brushes on the hydrogen terminated Si(100) substrates (Si-H substrates) was carried out via the surface initiated atom-transfer radical polymerization (ATRP) at room temperature. Thus, glycidyl methacrylate (GMA) polymer brushes were prepared by ATRP from the alpha bromoester functionalized Si-H surface. Kinetic studies revealed a linear increase in GMA polymer (PGMA) film thickness with reaction time, indicating that chain growth from the surface was a controlled "living" process. The graft polymerization proceeded more rapidly in the dimethylformamide/water (DMF/H(2)O) mixed solvent medium than in DMF, leading to much thicker PGMA growth on the silicon surface in the former medium. The chemical composition of the GMA graft polymerized silicon (Si-g-PGMA) surfaces were characterized by X-ray photoelectron spectroscopy (XPS). The fact that the epoxide functional groups of the grafted PGMA were preserved quantitatively was revealed in the reaction with ethylenediamine. The "living" character of the PGMA chain end was further ascertained by the subsequent growth of a poly(pentafluorostyrene) (PFS) block from the Si-g-PGMA surface, using the PGMA brushes as the macroinitiators. PMID- 15350106 TI - Synthesis, characterization, and adsorption properties of nanocrystalline ZSM-5. AB - Nanocrystalline ZSM-5 with a Si/Al ratio of 20 was synthesized using clear solutions and a hydrothermal synthesis procedure. The resulting ZSM-5 materials were characterized by powder X-ray diffraction, scanning electron microscopy (SEM), nitrogen adsorption isotherms, solid-state nuclear magnetic resonance, and toluene adsorption. A commercial ZSM-5 sample was similarly characterized for comparison with the synthesized materials. The particle sizes of the synthesized ZSM-5 samples were calculated using the measured external surface areas and were determined to be 15 and 60 nm. SEM images indicated that the ZSM-5 samples consist of agglomerated and possibly intergrown particles. Toluene adsorption measurements showed that the ZSM-5 sample with a particle size of 15 nm adsorbed approximately 50% more toluene than the other ZSM-5 samples, most likely due to the adsorption of toluene on the external surface. For the toluene adsorbed on the internal zeolite surface, approximately one toluene molecule was adsorbed per channel intersection for each of the ZSM-5 samples. PMID- 15350107 TI - Catalysis by hydrophobically modified poly(propylenimine) dendrimers having quaternary ammonium and tertiary amine functionality. AB - Four different quaternary ammonium chloride-modified poly(propylenimine) (PPI) dendrimers were synthesized by alkylation of a PPI dendrimer having eight dimethylamino end groups with 1-bromooctane or 1-bromododecane. By varying the mole ratio of alkyl bromide to dendrimer, averages of 4-10 quaternary ammonium groups were formed. The new amphiphilic dendrimers are surface active and are micellar catalysts in water. The dendrimers have critical aggregation concentrations between 8.5 x 10(-4) and 9.0 x 10(-5) M. Decarboxylation of 6 nitrobenzisoxazole-3-carboxylate at 25 degrees C was 650 times faster than in water alone in the presence of a dendrimer quaternized with eight dodecyl chains at a concentration of 2.45 mM in quaternary ammonium groups. The order of the catalytic efficiency of the new dendrimers decreased with the length and number of hydrophobic alkyl groups in the order (C(12))(8) > (C(12))(4) > (C(8))(10) > (C(8))(5). The pseudo-first-order rate constants for basic hydrolysis of p nitrophenyl hexanoate in pH 9.4 buffer at 30 degrees C using the (C(12))(8) and (C(12))(4) dendrimers were 26 and 13 times higher than those for hydrolysis with no dendrimer. The kinetic data were fit to a single-site binding model to evaluate the contributions of binding constants of reactants to the dendrimers and catalytic rate constants of the bound species to the overall catalytic activity. PMID- 15350108 TI - Patterned poly(N-isopropylacrylamide) brushes on silica surfaces by microcontact printing followed by surface-initiated polymerization. AB - Patterned poly(N-isopropylacrylamide) (PNIPAAm) brushes were fabricated on oxidized silicon wafers by surface-initiated atom transfer radical polymerization of N-isopropylacrylamide from a micropatterned initiator. The patterned surface initiator was prepared by microcontact-printing octadecyltrichlorosilane and backfilling with 3-(aminopropyl)triethoxysilane followed by amidization with 2 bromo-2-methylpropionic acid. XPS and FTIR confirmed the chemical structure of the surface initiator and the PNIPAAm brushes. Surface analysis techniques, including ellipsometry, contact angle goniometry, and X-ray reflectometry (XRR), were used to characterize the thickness, roughness, hydrophilicity, and density of the polymer brushes. Tapping-mode AFM imaging confirmed the successful patterning of the PNIPAAm brushes on the oxidized silicon substrates. Variable temperature ellipsometry indicated that the lower critical solution temperature of the hydrated PNIPAAm brush was broad, occurring over the range of 20-35 degrees C. A solvatochromic fluorophore, 6-propionyl-2-dimethylaminonaphthalene (Prodan), in the PNIPAAm brush layers yielded a very similar emission to that in DMF, which can be attributed to the similarity of their chemical structures. Fluorescence microscopy further proved the successful patterning of the polymer brushes and suggested that the Prodan is localized in the patterned PNIPAAm brushes and excluded from the surrounding octadecyltrichlorosilane regions. PMID- 15350109 TI - Porphyrinphosphonate fibers on mica and molecular rows on graphite. AB - meso-Tetra(phenyl-p-phosphonate) porphyrin forms rigid and well-separated fibers of monomolecular thickness (2.8 nm) and lengths of several micrometers on mica at pH 13 (octasodium salt). The formation of these fibers could be observed directly by tapping mode scanning force microscopy (SFM) and was induced by capillary forces. Normal height images or images with a topographical inversion were observed depending on the distance of the SFM tip. Amplitude-distance curves indicated that a stable meniscus was formed on hydrophilic surface areas below a tip-sample separation of 20 nm. The meniscus let the original nanorods appear as ditches in the mica surface and enabled rearrangements. A partly protonated form of the same porphyrin (pH 11.5) gave rows of flat-lying porphyrins on graphite, which appear with molecular resolution in SFM images as well as two-dimensional platelets of monomolecular thickness. PMID- 15350110 TI - Thermal reduction of Pd molecular cluster precursors at highly ordered pyrolytic graphite surfaces. AB - Highly ordered pyrolytic graphite (HOPG) surfaces were modified by the adsorption of Pd molecular precursors from solution. Two palladium-containing molecular precursors were studied, a mononuclear one and a trinuclear one, to compare their affinities and distributions at substrate surfaces. To obtain Pd nanoparticles, these neutral molecular precursors were reduced under a hydrogen atmosphere. Thermogravimetric analysis was carried out to establish the behavior of these precursors at various temperatures. Understanding the thermal stability of these compounds is very important to establish the appropriate conditions to form metallic Pd. The modified surface has been characterized by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy; also, the reductive process was monitored by XPS. Remarkable differences were observed between the mononuclear and trinuclear compounds in terms of dispersion, particle size, and homogeneity. The preference of the trinuclear compound was to deposit at HOPG defects, in contrast to that of the mononuclear one, which was agglomeration on all surfaces. After the application of this technique, not only Pd nanoparticles but also Pd nanowires were obtained. PMID- 15350111 TI - Optimization of dye-doped silica nanoparticles prepared using a reverse microemulsion method. AB - Fluorescent labeling based on silica nanoparticles facilitates unique applications in bioanalysis and bioseparation. Dye-doped silica nanoparticles have significant advantages over single-dye labeling in signal amplification, photostability and surface modification for various biological applications. We have studied the formation of tris(2,2'-bipyridyl)dichlororuthenium(II) (Ru(bpy)) dye-doped silica nanoparticles by ammonia-catalyzed hydrolysis of tetraethyl orthosilicate (TEOS) in water-in-oil microemulsion. The fluorescence spectra, particle size, and size distribution of Ru(bpy) dye-doped silica nanoparticles were examined as a function of reactant concentrations (TEOS and ammonium hydroxide), nature of surfactant molecules, and molar ratios of water to surfactant (R) and cosurfactant to surfactant (p). The particle size and fluorescence spectra were dependent upon the type of microemulsion system chosen. The particle size was found to decrease with an increase in concentration of ammonium hydroxide and increase in water to surfactant molar ratio (R) and cosurfactant to surfactant molar ratio (p). This optimization study of the preparation of dye-doped silica nanoparticles provides a fundamental knowledge of the synthesis and optical properties of Ru(bpy) dye-doped silica nanoparticles. With this information, these nanoparticles can be easily manipulated, with regard to particle size and size distribution, and bioconjugated as needed for bioanalysis and bioseparation applications. PMID- 15350112 TI - Comparison study of the solution phase versus solid phase place exchange reactions in the controlled functionalization of gold nanoparticles. AB - Gold nanoparticles offer tremendous potential in the areas of nanoelectronics, bio- and chemosensors, and catalysis. However, before these applications are realized, the surface functionality of nanoparticles must be better controlled. Our lab has recently reported a novel synthetic approach for making monofunctionalized nanoparticles through a solid phase place exchange reaction. Monofunctionalized gold nanoparticles may also be prepared through a solution phase place exchange reaction. In this study, we compared the efficiency of these two separate approaches toward controlled functionalization of gold nanoparticles by (1)H NMR, Fourier transform infrared (FT-IR), and transmission electron microscopy (TEM) analysis. We found that the solid phase place exchange approach is much more efficient at producing monofunctionalized gold nanoparticles. (1)H NMR data were used to give a semiquantitative count of substituted bifunctional ligands, and FT-IR spectra supported these findings. Furthermore, we used a diamine coupling reaction of nanoparticles to show the presence of single or multiple functional groups on the nanoparticle surface by TEM analysis. PMID- 15350113 TI - Ultrasound-assisted polyol method for the preparation of SBA-15-supported ruthenium nanoparticles and the study of their catalytic activity on the partial oxidation of methane. AB - Metallic Ru nanoparticles have been successfully produced and incorporated into the pores of SBA-15 in situ employing a simple ultrasound-assisted polyol method. The product has been confirmed by X-ray diffraction, transmission electron microscopy, and X-ray photoelectron spectroscopy, where ultrasound provides both the energy for the reduction of the Ru(III) ion and the driving force for the loading of the Ru(0) nanoparticles into the SBA-15 pores. An ultrasound-assisted insertion mechanism has been proposed based on the microjets and shake-wave effect of the collapsed bubbles. The catalytic properties of the SBA-15-supported Ru nanoparticles have been tested by the partial oxidization of methane and show very high activity and high CO selectivity. PMID- 15350114 TI - Nanoparticles of varying hydrophobicity at the emulsion droplet-water interface: adsorption and coalescence stability. AB - The coalescence stability of poly(dimethylsiloxane) emulsion droplets in the presence of silica nanoparticles ( approximately 50 nm) of varying contact angles has been investigated. Nanoparticle adsorption isotherms were determined by depletion from solution. The coalescence kinetics (determined under coagulation conditions at high salt concentration) and the physical structure of coalesced droplets were determined from optical microscopy. Fully hydrated silica nanoparticles adsorb with low affinity, reaching a maximum surface coverage that corresponds to a close packed monolayer, based on the effective particle radius and controlled by the salt concentration. Adsorbed layers of hydrophilic nanoparticles introduce a barrier to coalescence of approximately 1 kT, only slightly reduce the coalescence kinetics, and form kinetically unstable networks at high salt concentrations. Chemically hydrophobized silica nanoparticles, over a wide range of contact angles (25 to >90 degrees ), adsorb at the droplet interface with high affinity and to coverages equivalent to close-packed multilayers. Adsorption isotherms are independent of the contact angle, suggesting that hydrophobic attraction overcomes electrostatic repulsion in all cases. The highly structured and rigid adsorbed layers significantly reduce coalescence kinetics: at or above monolayer surface coverage, stable flocculated networks of droplets form and, regardless of their wettability, particles are not detached from the interface during coalescence. At sub-monolayer nanoparticle coverages, limited coalescence is observed and interfacial saturation restricts the droplet size increase. When the nanoparticle interfacial coverage is >0.7 and <1.0, mesophase-like microstructures have been noted, the physical form and stability of which depends on the contact angle. Adsorbed nanoparticle layers at monolayer coverage and composed of a mixture of nanoparticles with different hydrophobisation levels form stable networks of droplets, whereas mixtures of hydrophobized and hydrophilic nanoparticles do not effectively stabilize emulsion droplets. PMID- 15350115 TI - Porphyrin-sensitized nanoparticulate TiO2 as the photoanode of a hybrid photoelectrochemical biofuel cell. AB - Porphyrin-sensitized nanoparticulate TiO(2) on conducting glass has been investigated as a photoanode material for a new cell that converts light energy into electricity. The cell is a hybrid of a dye-sensitized nanoparticulate semiconductor photoelectrochemical solar cell, and a biofuel cell that oxidizes glucose. Porphyrin molecules excited by light inject electrons into the photoanode, from where they enter the external circuit. The resulting porphyrin radical cations are reduced by NADH in aqueous buffer, ultimately regenerating the photoanode and producing NAD(+). Glucose dehydrogenase oxidizes glucose, and in the process recycles NAD(+) back to NADH. The photoanode is coupled with a suitable cathode to make a functioning cell (Hg/Hg(2)SO(4) was employed for evaluation purposes). The cell produces 1.1 V at open circuit and has a fill factor of 0.61. These values are both significantly higher than those for a previously reported cell of a similar type based on an SnO(2) electrode. PMID- 15350116 TI - Reactions of antimicrobial species to imidazole-microwave plasma reacted poly(dimethylsiloxane) surfaces. AB - Microwave plasma reactions of imidazole, 2-methylimidazole, and 2-ethylimidazole on poly(dimethylsiloxane) (PDMS) surfaces resulted in the formation of species containing conjugated surface domains which can be utilized for further reactions. When imidazole and its derivatives were used, polymerization of imidazole and the formation of C=C and CN conjugated species occurred. However, the extent of reactions for each monomer depends on not only the reaction time but also the molecular structure. For methyl- and ethyl-substituted imidazole, more stable radical species are generated and sustain their excited state in the high-energy plasma environments. Specifically, dehydrogenated 2-methyl, 2 ethylimidazole radicals and (*)N=CR-NH(*) (R = -CH(3), -CH(2)CH(3)) species exhibit higher stability than dehydrogenated imidazole radicals and (*)N=CH-NH(*) species under plasma reaction conditions. Such prepared surfaces are capable of attaching antimicrobial drugs via the Pinner synthesis. These studies show that it is possible to react antimicrobial species such as chloramphenicol, and this promising approach offers numerous applications of microwave plasma reactions in biotechnology. Quantitative analysis of the depth of surface reactions was accomplished by using variable angle ATR FT-IR spectroscopy. PMID- 15350117 TI - Vertical and lateral order in adsorbed water layers on anatase TiO2(101). AB - The structure and energetics of thin water overlayers on the (101) surface of TiO(2)-anatase have been studied through first-principles molecular dynamics simulations at T = 160 K. At one monolayer coverage, H(2)O molecules are adsorbed at the 5-fold Ti sites (Ti(5c)), forming an ordered crystal-like 2D layer with no significant water-water interactions. For an adsorbed bilayer, H(2)O molecules at both Ti(5c) and bridging oxygen (O(2c)) sites form a partially ordered structure, where the water oxygens occupy regular sites but the orientation of the molecules is disordered; in addition, stress-relieving defects are usually present. When a third layer is adsorbed, very limited parallel and perpendicular order is observed above the first bilayer. The calculated energetics of multilayer adsorption is in good agreement with recent temperature-programmed desorption data. PMID- 15350118 TI - Liquid coating of moving fiber at the nanoscale. AB - Using large scale molecular dynamics, we study the contact line motion of a liquid meniscus crossed by a moving nanofiber. Varying the amplitude of the liquid/solid interactions, we analyze the shape of the meniscus versus time for a range of velocities. The associated contact angles are estimated by fitting the profiles using the James equation. The corresponding flux lines describing the displacement of the liquid molecules inside the meniscus have also been measured. The analysis of the dynamic contact angle is in agreement with the molecular kinetic theory and confirms the existence of an optimal speed for wetting. PMID- 15350119 TI - van der Waals interaction between internal aqueous droplets and the external aqueous phase in double emulsions. AB - A mathematical model for analyzing the van der Waals interaction between the internal aqueous droplets (W(1)) and the external aqueous phase (W(2)) of double emulsions has been established. The effects of Hamaker constants of the materials forming the system, especially those of the two different adsorbed surfactant layers with uniform density (A(1) and A(2)), on the van der Waals interaction were investigated. The overall van der Waals interaction across the oil film is a combined result of four individual parts, that is, W(1)-W(2), A(1)-A(2), W(1) A(1), and A(2)-W(2) van der Waals interaction, and it may be either attractive or repulsive depending on many factors. It was found that the overall van der Waals interaction is dominated by the W(1)-W(2) interaction at large separation distances between the W(1)/O and O/W(2) interfaces, while it is mostly determined by the A(1)-A(2) interaction when the two interfaces are extremely close. Specifically, in the cases when the value of the Hamaker constant of the oil phase is intermediate between those of W(1) and W(2) and there is a thick oil film separating the two interfaces, a weak repulsive overall van der Waals interaction will prevail. If the Hamaker constant of the oil phase is intermediate between those of A(1) and A(2) and the two interfaces are very close, the overall van der Waals interaction will be dominated by the strong repulsive A(1)-A(2) interaction. The repulsive van der Waals interaction at such cases helps stabilize the double emulsions. PMID- 15350120 TI - Microemulsion synthesis of fluorinated latex spheres. PMID- 15350121 TI - Microcapsule assembly of human serum albumin at the liquid/liquid interface by the pendent drop technique. PMID- 15350122 TI - Surfactant-assisted solvothermal synthesis of Co3O4 hollow spheres with oriented aggregation nanostructures and tunable particle size. PMID- 15350123 TI - New insights into the mechanisms of protein misfolding and aggregation in amyloidogenic diseases derived from pressure studies. AB - Hydrostatic pressure is a robust tool for studying the thermodynamics of protein folding and protein interactions, as well as the dynamics and structure of folding intermediates. One of the main innovations obtained from using high pressure is the stabilization of folding intermediates such as molten-globule conformations, thus providing a unique opportunity for characterizing their structure and dynamics. Equally important is the prospect of understanding protein misfolding diseases by using pressure to populate partially folded intermediates at the junction between productive and off-pathway folding, which may give rise to misfolded proteins, aggregates, and amyloids. High hydrostatic pressure (HHP) has also been used to dissociate nonamyloid aggregates and inclusion bodies. In many proteins, the competition between correct folding and misfolding can lead to formation of insoluble aggregates, an important problem for the biotechnology industry and for human pathologies such as amyloidosis, Alzheimer's, Parkinson's, prion's, and tumor diseases. The diversity of diseases that result from protein misfolding has made this theme an important research focus for pharmaceutical and biotechnology companies. The use of high-pressure promises to contribute to the identification of the mechanisms behind these defects and creation of therapies against these diseases. PMID- 15350124 TI - Structure of the Mg2+-loaded C-lobe of cardiac troponin C bound to the N-domain of cardiac troponin I: comparison with the Ca2+-loaded structure. AB - Cardiac troponin C (cTnC) is the Ca(2+)-binding component of the troponin complex and, as such, is the Ca(2+)-dependent switch in muscle contraction. This protein consists of two globular lobes, each containing a pair of EF-hand metal-binding sites, connected by a linker. In the N lobe, Ca(2+)-binding site I is inactive and Ca(2+)-binding site II is primarily responsible for initiation of muscle contraction. The C lobe contains Ca(2+)/Mg(2+)-binding sites III and IV, which bind Mg(2+) with lower affinity and play a structural as well as a secondary role in modulating the Ca(2+) signal. To understand the structural consequences of Ca(2+)/Mg(2+) exchange in the C lobe, we have determined the NMR solution structure of the Mg(2+)-loaded C lobe, cTnC(81-161), in a complex with the N domain of cardiac troponin I, cTnI(33-80), and compared it with a refined Ca(2+) loaded structure. The overall tertiary structure of the Mg(2+)-loaded C lobe is very similar to that of the refined Ca(2+)-loaded structure as evidenced by the root-mean-square deviation of 0.94 A for all backbone atoms. While metal dependent conformational changes are minimal, substitution of Mg(2+) for Ca(2+) is characterized by condensation of the C-terminal portion of the metal-binding loops with monodentate Mg(2+) ligation by the conserved Glu at position 12 and partial closure of the cTnI hydrophobic binding cleft around site IV. Thus, conformational plasticity in the Ca(2+)/Mg(2+)-dependent binding loops may represent a mechanism to modulate C-lobe cTnC interactions with the N domain of cTnI. PMID- 15350125 TI - Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2). AB - The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. Subsequent site-directed mutagenesis of the predicted binding domain further validated the model. This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids. PMID- 15350126 TI - Effect of salt on the urea-unfolded form of barstar probed by m value measurements. AB - To probe for residual structure present in the urea-unfolded form of the small protein barstar, to determine how salt might modulate such structure, and to determine how such structure might affect the stability of the protein, mutant variants that display m values different from that of the wild-type protein have been studied. The mutant proteins were obtained by site-directed mutagenesis at residue positions located on the surface of the folded protein. The m value, which represents the preferential free energy of interaction of urea with the unfolded form in comparison to that with the folded state, was determined from equilibrium urea-induced unfolding curves. Mutant proteins for which the m values were significantly greater than (m(+) mutant forms), significantly smaller than (m(-) mutant forms), or similar to (m(0) mutant forms) the m value determined for the wild-type protein were studied. The unfolded forms of the m(0), m(+) and m(-) mutant proteins represent different components within the unfolded form ensemble, which differ from each other in their solvent-exposed surface areas. Hence, the m value has been used as a measure of residual structure in the unfolded form. To further understand the nature of structures present in the unfolded form ensemble, the effects of the salt KCl on the stabilities of the wild-type and the mutant proteins, as well as on the structures present in the unfolded form ensemble, were also studied. It was found that the m values of the m(0), m(+) and m(-) mutant proteins all converge to the wild-type m value in the presence of KCl. This result indicates that the salt modulates residual structure in the unfolded form by screening electrostatic interactions that maintain compact and expanded components in the unfolded protein ensemble. The use of free energy cycles has allowed the effect of salt on the structure and free energy of the unfolded protein to be related to the stability of the protein. PMID- 15350127 TI - Crystal structure of phenylalanine ammonia lyase: multiple helix dipoles implicated in catalysis. AB - The first three-dimensional structure of phenylalanine ammonia lyase (PAL) has been determined at 2.1 A resolution for PAL from Rhodosporidium toruloides. The enzyme is structurally similar to the mechanistically related histidine ammonia lyase (HAL), with PAL having an additional approximately 160 residues extending from the common fold. We propose that catalysis (including lowering the pK(a) of nonacidic C3 of l-phenylalanine for an E1cb mechanism) is potentially governed by dipole moments of seven alpha helices associated with the PAL active site (six positive poles and one negative pole). Cofactor 3,5-dihydro-5-methylidene-4H imidazol-4-one (MIO) resides atop the positive poles of three helices, for increasing its electrophilicity. The helix dipoles appear fully compatible with a model of phenylalanine docked in the active site of PAL having the first covalent bond formed between the amino group of substrate and the methylidene group of MIO: 12 highly conserved residues (near the N termini of helices for enhancing function) are poised to serve roles in substrate recognition, MIO activation, product separation, proton donation, or polarizing electrons from the phenyl ring of substrate for activation of C3; and a highly conserved His residue (near the C terminus of the one helix that directs its negative pole toward the active site to increase the residue's basicity) is positioned to act as a general base, abstracting the pro-S hydrogen from C3 of substrate. A similar mechanism is proposed for HAL, which has a similar disposition of seven alpha helices and similar active-site residues. The helix dipoles appear incompatible with a proposed mechanism that invokes a carbocation intermediate. PMID- 15350128 TI - Crystal structure of quinone reductase 2 in complex with resveratrol. AB - Resveratrol has been shown to have chemopreventive, cardioprotective, and antiaging properties. Here, we report that resveratrol is a potent inhibitor of quinone reductase 2 (QR2) activity in vitro with a dissociation constant of 35 nM and show that it specifically binds to the deep active-site cleft of QR2 using high-resolution structural analysis. All three resveratrol hydroxyl groups form hydrogen bonds with amino acids from QR2, anchoring a flat resveratrol molecule in parallel with the isoalloxazine ring of FAD. The unique active-site pocket in QR2 could potentially bind other natural polyphenols such as flavonoids, as proven by the high affinity exhibited by quercetin toward QR2. K562 cells with QR2 expression suppressed by RNAi showed similar properties as resveratrol treated cells in their resistance to quinone toxicity. Furthermore, the QR2 knockdown K562 cells exhibit increased antioxidant and detoxification enzyme expression and reduced proliferation rates. These observations could imply that the chemopreventive and cardioprotective properties of resveratrol are possibly the results of QR2 activity inhibition, which in turn, up-regulates the expression of cellular antioxidant enzymes and cellular resistance to oxidative stress. PMID- 15350129 TI - Crystallographic analysis of the Pseudomonas aeruginosa strain K122-4 monomeric pilin reveals a conserved receptor-binding architecture. AB - Adherence of pathogens to host cells is critical for the initiation of infection and is thus an attractive target for anti-infective therapeutics and vaccines. In the opportunistic human pathogen Pseudomonas aeruginosa, host-cell adherence is achieved predominantly by type IV pili. Analysis of several clinical strains of P. aeruginosa reveals poor sequence conservation between pilin genes, including the residues in the receptor-binding site. Interestingly, the receptor-binding sites appear to retain a conserved surface epitope because all Pseudomonas type IV pili recognize the same receptor on the host cell and cross-reactive antibodies specific for the receptor-binding site exist. Here, we present the crystallographic analysis of two crystal forms of truncated pilin from P. aeruginosa strain K122-4 (DeltaK122-4) at 1.54 and 1.8 A resolution, respectively. The DeltaK122-4 structure is compared to other crystallographically determined type IV pilin structures and an NMR structure of DeltaK122-4 pilin. A comparison with the structure of the highly divergent P. aeruginosa strain K (DeltaPAK) pilin indicates that the receptor-binding loop in both pilins forms a shallow depression with a surface that is formed by main-chain atoms. Conservation of this putative binding site is independent of the sequence as long as the main-chain conformation is conserved and could therefore explain the shared receptor specificity and antibody cross reactivity of highly divergent Pseudomonas type IV pilins. PMID- 15350130 TI - Entropy effects on protein hinges: the reaction catalyzed by triosephosphate isomerase. AB - Many proteins utilize segmental motions to catalyze a specific reaction. The Omega loop of triosephosphate isomerase (TIM) is important for preventing the loss of the reactive enediol(ate) intermediate. The loop opens and closes even in the absence of the ligand, and the loop itself does not change conformation during movement. The conformational changes are localized to two hinges at the loop termini. Glycine is never observed in native TIM hinge sequences. In this paper, the hypothesis that limited access to conformational space is a requirement for protein hinges involved in catalysis was tested. The N-terminal hinge was mutated to P166/V167G/W168G (PGG), and the C-terminal hinge was mutated to K174G/T175G/A176G (GGG) in chicken TIM. The single-hinge mutants PGG and GGG had k(cat) values 200-fold lower than that of the wild type and K(m) values 10 fold higher. The k(cat) of double-hinge mutant P166/V167G/W168G/K174G/T175G/A176G was reduced 2500-fold; the K(m) was 10-fold higher. A combination of primary kinetic isotope effect measurements, isothermal calorimetric measurements, and (31)P NMR spectroscopic titration with the inhibitor 2-phosphoglycolate revealed that the mutants have a different ligand-binding mode than that of the wild-type enzyme. The predominant conformations of the mutants even in the presence of the inhibitor are loop-open conformations. In conclusion, mutation of the hinge residues to glycine resulted in the sampling of many more hinge conformations with the consequence that the population of the active-closed conformation is reduced. This reduced population results in a reduced catalytic activity. PMID- 15350131 TI - Quantitative analysis of a RNA-cleaving DNA catalyst obtained via in vitro selection. AB - In vitro selections performed in the presence of Mg(2+) generated DNA sequences capable of cleaving an internal ribonucleoside linkage. Several of these, surprisingly, displayed intermolecular catalysis and catalysis independent of Mg(2+), features that the selection protocol was not explicitly designed to select. A detailed physical organic analysis was applied to one of these DNAzymes, termed 614. First, the progress curve for the reaction was dissected to identify factors that prevented the molecule from displaying clean first-order transformation kinetics and 100% conversion. Several factors were identified and quantitated, including (a) competitive intra- and intermolecular rate processes, (b) alternative reactive and unreactive conformations, and (c) mutations within the catalyst. Other factors were excluded, including "approach to equilibrium" kinetics and product inhibition. The possibility of complementary strand inhibition was demonstrated but was shown to not be a factor under the conditions of these experiments. The rates of the intra- and intermolecular processes were compared, and saturation models for the intermolecular process were built. The rate-limiting step for the intermolecular reaction was found to be the association/folding of the enzyme with the substrate and not the cleavage step. The DNAzyme 614 is more active in trans than in cis and more active at temperatures below the selection temperature than at the selection temperature. Many of these properties have not been reported in similar systems; these results therefore expand the phenomenology known for this class of DNA-based catalysts. A brief survey of other catalysts arising from this selection found other Mg(2+) independent DNAzymes and provided a preliminary view of the ruggedness of the landscape, relating function to structure in sequence space. Hypotheses are suggested to account for the fact that a selection in the presence of Mg(2+) did not exploit this Mg(2+). This study of a specific catalytically active DNAzyme is an example of studies that will be necessary generally to permit in vitro selection to help us understand the distribution of function in sequence space. PMID- 15350132 TI - The mechanism of target base attack in DNA cytosine carbon 5 methylation. AB - We measured the tritium exchange reaction on cytosine C(5) in the presence of AdoMet analogues to investigate the catalytic mechanism of the bacterial DNA cytosine methyltransferase M.HhaI. Poly(dG-dC) and poly(dI-dC) substrates were used to investigate the function of the active site loop (residues 80-99), stability of the extrahelical base, base flipping mechanism, and processivity on DNA substrates. On the basis of several experimental approaches, we show that methyl transfer is the rate-limiting pre-steady-state step. Further, we show that the active site loop opening contributes to the rate-limiting step during multiple cycles of catalysis. Target base activation and nucleophilic attack by cysteine 81 are fast and readily reversible. Thus, the reaction intermediates involving the activated target base and the extrahelical base are in equilibrium and accumulate prior to the slow methyl transfer step. The stability of the activated target base depends on the active site loop closure, which is dependent on the hydrogen bond between isoleucine 86 and the guanine 5' to the target cytosine. These interactions prevent the premature release of the extrahelical base and uncontrolled solvent access; the latter modulates the exchange reaction and, by implication, the mutagenic deamination reaction. The processive catalysis by M.HhaI is also regulated by the interaction between isoleucine 86 and the DNA substrate. Nucleophilic attack by cysteine 81 is partially rate limiting when the target base is not fully stabilized in the extrahelical position, as observed during the reaction with the Gln(237)Trp mutant or in the cytosine C(5) exchange reaction in the absence of the cofactor. PMID- 15350133 TI - Matrix metalloproteinase triple-helical peptidase activities are differentially regulated by substrate stability. AB - Matrix metalloproteinases (MMPs) are involved in physiological remodeling as well as pathological destruction of tissues. The turnover of the collagen triple helical structure has been ascribed to several members of the MMP family, but the determinants for collagenolytic specificity have not been identified. The present study has compared the triple-helical peptidase activities of MMP-1 and MMP-14 (membrane-type 1 MMP; MT1-MMP). The ability of each enzyme to efficiently hydrolyze the triple helix was quantified using chemically synthesized fluorogenic triple-helical substrates that, via addition of N-terminal alkyl chains, differ in their thermal stabilities. One series of substrates was modeled after a collagenolytic MMP consensus cleavage site from types I-III collagen, while the other series had a single substitution in the P(1)' subsite of the consensus sequence. The substitution of Cys(4-methoxybenzyl) for Leu in the P(1)' subsite was greatly favored by MMP-14 but disfavored by MMP-1. An increase in substrate triple-helical thermal stability led to the decreased ability of the enzyme to cleave such substrates, but with a much more pronounced effect for MMP 1. Increased thermal stability was detrimental to enzyme turnover of substrate (k(cat)), but not binding (K(M)). Activation energies were considerably lower for MMP-14 hydrolysis of triple-helical substrates compared with MMP-1. Overall, MMP 1 was found to be less efficient at processing triple-helical structures than MMP 14. These results demonstrate that collagenolytic MMPs have subtle differences in their abilities to hydrolyze triple helices and may explain the relative collagen specificity of MMP-1. PMID- 15350134 TI - Foot-and-mouth disease virus leader proteinase: specificity at the P2 and P3 positions and comparison with other papain-like enzymes. AB - The foot-and-mouth disease virus Leader proteinase (L(pro)) frees itself from the growing viral polyprotein by self-processing between its own C-terminus and the N terminus of the subsequent protein VP4. The ArgLysLeuLys*GlyAlaGlyGln sequence is recognized. The proteinase subsequently cleaves the two isoforms of host cell protein eukaryotic initiation factor (eIF) 4G at the AlaAsnLeuGly*ArgThrThrLeu (eIF4GI) and LeuAsnValGly*SerArgArgSer (eIF4GII) sequences. The enzyme does not, however, recognize the sequence on eIF4GII (AlaAspPheGly*ArgGlnThrPro) which is analogous to that recognized on eIF4GI. To investigate the basis for this specificity, we used site-directed mutagenesis to show that the presence of Phe at the P2 position or Asp at the P3 position severely compromises self processing. Furthermore, these substitutions also give rise to the production of aberrant cleavage products. As Leu is the preferred amino acid at P2, the specificity of L(pro) is reminiscent of that of cathepsin K. This cellular proteinase can also process collagen through its ability to accept proline at the P2 position. Investigation of the L(pro) substrate specificity showed, however, that in contrast to cathepsin K, L(pro) cannot accept Pro at P2 and does not cleave collagen. Subtle variations in the arrangement of the S2 binding pockets on the enzymes are responsible for these differences in specificity. PMID- 15350135 TI - Distances between tropomyosin sites across the muscle thin filament using luminescence resonance energy transfer: evidence for tropomyosin flexibility. AB - To obtain information about the interaction of tropomyosin (Tm) with actin associated with the regulatory states of the muscle thin filament, we used luminescence resonance energy transfer (LRET) between Tb(3+) as a donor and rhodamine as an acceptor. A novel Tb(3+) chelator, S-(2-nitro-5 thiobenzoate)cysteaminyl-DTPA-Cs124, was synthesized, which specifically labels Cys groups in proteins. With the Tb chelate as the donor and tetramethylrhodamine 5-maleimide as the acceptor, both bound to specific Cys groups of Tm, we obtained 67 A as the distance between Tm's across the actin filament, a much shorter value than that obtained from structural studies (72-86 A). The difference appears to be due to submillisecond motion associated with Tm flexibility, which brings the probes closer during the millisecond lifetime of the donor. Ca(2+) did not change the energy transfer with the reconstituted thin filament, but myosin subfragment 1 decreased the transfer, consistent with either a 5-6 A increase in distance or, more likely, a decrease in flexibility. PMID- 15350136 TI - In vivo posttranslational modifications of the high mobility group A1a proteins in breast cancer cells of differing metastatic potential. AB - The high mobility group (HMG) proteins are important modulators of chromatin structure and gene transcription. Overexpression of HMGA1 proteins in vivo induces neoplastic transformation and promotes a highly metastatic cellular phenotype. This study focuses on characterization of HMGA1a in vivo posttranslational modification (PTM) patterns found in a nonmetastatic and two metastatic lines of MCF-7 human breast cancer cells of differing tumorigenic potential. PTM types and the amino acids on which they occur were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. Mass analysis was restricted to MALDI peaks having less than +/-150 parts per million (ppm) error, thereby holding our analysis to a more stringent criterion than previously published work with the HMG proteins. Validation of MALDI-TOF MS analysis was accomplished utilizing electrospray ionization tandem mass spectrometry (ESI MS/MS) and manual analysis of ion fragmentation spectra. Patterns and sites of PTMs identified in this study suggest that HMGA1a proteins, like the histones, exhibit a biochemical modification "code" that relates to cellular function. For example, both increased levels of acetylation and a previously unidentified dimethylation of both lysine and arginine residues were found on HMGA1a proteins from metastatic cells compared to proteins found in their nonmetastatic precursors. Additionally, the types of modification present on lysine-45 (e.g., unmodified, acetylation, or dimethylation) varied, depending on the metastatic potential of cells. These findings suggest that examination of the PTM patterns on HMGA1 proteins may provide valuable information concerning the physiological and phenotypic state of mammalian cells. PMID- 15350137 TI - Identification of a binding domain of the endothelin-B receptor using a selective IRL-1620-derived photoprobe. AB - On the basis of the structure of IRL-1620, a specific agonist of the endothelin-B receptor subtype (ET(B)), a few photosensitive analogues were developed to investigate the binding domain of the receptor. Among those, a derivative containing the photoreactive amino acid, p-benzoyl-l-phenylalanine in position 5 showed, as assessed with endothelin-A (ET(A)) and ET(B) receptor paradigms, pharmacological properties very similar to those of IRL-1620. The binding capacity of the probe was also evaluated on transfected Chinese hamster ovary (CHO) cells overexpressing the human ET(B) receptor. Data showed that binding of the radiolabeled peptide was inhibited by ET-1 and IRL-1620. Therefore, this photolabile probe was used to label the ET(B) receptor found in CHO cells. Photolabeling produced a ligand-protein complex appearing on SDS-PAGE at around 49 kDa. An excess of ET-1 or IRL-1620 completely abolished the formation of the complex, showing the selectivity of the photoprobe. Digestions of the [Bpa(5),Tyr((125)I)(6)]IRL-1620-ET(B) complex were carried out, and receptor fragments were analyzed to define the region of the receptor where the ligand interacts. Results showed that Endo Lys-C digestion gave a 3.8-kDa fragment corresponding to the Asp(274)-Lys(303) segment, whereas migration after V8 digestion revealed a fragment of 4.6 kDa. Because the fragments of these two digestions must overlap, the latter would be the Trp(275)-Asp(313) stretch. A cleavage with CNBr confirmed the identity of the binding domain by giving a fragment of 3.6 kDa, corresponding to Gln(267)-Met(296). Thus, the combined cleavage data strongly suggested that the agonist binding domain of ET(B) includes a portion of the fifth transmembrane domain, between residues Trp(275) and Met(296). PMID- 15350138 TI - Heat-induced secondary structure and conformation change of bovine serum albumin investigated by Fourier transform infrared spectroscopy. AB - Fourier transform infrared (FT-IR) spectra were measured for an aqueous solution (pD = 5.40) of defatted monomer bovine serum albumin (BSA) over a temperature range of 25-90 degrees C to investigate temperature-induced secondary structure and conformation changes. The curve fitting method combined with the Fourier self deconvolution technique allowed us to explore details of the secondary structure and conformation changes in defatted BSA. Particularly striking in the FT-IR spectra was an observation of the formation of an irreversible intermolecular beta-sheet of BSA on heating above 70 degrees C. A band at 1630 cm(-1) in the spectra was assigned to short-segment chains connecting alpha-helical segments. The transition temperature for the short-segment chains connecting alpha-helical segments is lower by 17-18 degrees C, when compared to those of the alpha-helix, turn, and intermolecular beta-sheet structures of BSA, suggesting that the alpha helix and turn structures of BSA are cooperatively denatured on heating. Moreover, the results give an important feature in heat-induced denaturation of BSA that the conformation changes occur twice around both 57 and 75 degrees C. The appearance of two peaks is interpreted by the collapse of the N-terminal BSA domain due to the crevice in the vicinity between domains I and II at low temperature transition and by the change in cooperative unit composed of the other two BSA domains at high-temperature transition. PMID- 15350139 TI - Synthesis and characterization of biotinylated forms of insulin-like growth factor-1: topographical evaluation of the IGF-1/IGFBP-2 AND IGFBP-3 interface. AB - Activation of the insulin-like growth factor-1 (IGF)-1 receptor signaling pathways by IGF-1 and IGF-2 results in mitogenic and anabolic effects. The bioavailability of the IGFs is regulated by six soluble binding proteins, the insulin-like growth factor binding proteins (IGFBPs), which bind with approximately 0.1 nM affinity to the IGFs and often serve as endogenous antagonists of IGF action. To identify key domains of IGF-1 involved in the interaction with IGFBP-2 and IGFBP-3, we employed IGF-1 selectively biotinylated on residues Gly 1, Lys 27, Lys 65, and Lys 68. All monobiotinylated species of IGF-1 exhibited high affinity ( approximately 0.1-0.2 nM) for IGFBP-2 and IGFBP-3 in solid-phase-binding assays. However, different labeling intensities were observed in ligand blot analysis of IGFBP-2 and IGFBP-3. The N(epsilon)(Lys65/68)(biotin)-IGF-1 (N(epsilon)(Lys65/68b)-IGF-1) probe exhibited the highest signal intensity, while N(alpha)(Gly1b)-IGF-1 and N(epsilon)(Lys27b) IGF-1 demonstrated significantly lower signals. When taken together, these results suggest that, once bound to IGFBP-2 or IGFBP-3, the biotin moieties of N(alpha)(Gly1b)-IGF-1 and N(epsilon)(Lys27b)-IGF-1 are inaccessible to NeutrAvidin-peroxidase, the secondary binding component. Ligand blots using IGF-1 derivatized with a long chain form of the N-hydroxysuccinimide biotin (NHS biotin) to yield N(alpha)(Gly1)(LC-biotin)-IGF-1 and N(epsilon)(Lys27)(LC-biotin) IGF-1 demonstrated increased signal intensity compared with their NHS-biotin counterparts. In BIAcore analysis, IGFBP-2 and IGFBP-3 bound only to the N(epsilon)(Lys65/68b)-IGF-1-coated flowcell of a biosensor chip, confirming the inaccessibility of Gly 1 and Lys 27 when IGF-1 is bound to IGFBP-2 and IGFBP-3. These data confirm the involvement of the IGFBP-binding domain on IGF-1 in binding to IGFBP-2 and IGFBP-3 and support involvement of the IGF-1R-binding domain in IGFBP binding. PMID- 15350140 TI - Kinetic evaluation of catalase and peroxygenase activities of tyrosinase. AB - Tyrosinase is a copper monooxygenase containing a coupled dinuclear copper active site (type-3 copper), which catalyzes oxygenation of phenols (phenolase activity) as well as dehydrogenation of catechols (catecholase activity) using O(2) as the oxidant. In this study, catalase activity (conversion of H(2)O(2) to (1/2)O(2) and H(2)O) and peroxygenase activity (H(2)O(2)-dependent oxygenation of substrates) of mushroom tyrosinase have been examined kinetically by using amperometric O(2) and H(2)O(2) sensors. The catalase activity has been examined by monitoring the initial rate of O(2) production from H(2)O(2) in the presence of a catalytic amount of tyrosinase in 0.1 M phosphate buffer (pH 7.0) at 25 degrees C under initially anaerobic conditions. It has been found that the catalase activity of mushroom tyrosinase is three-order of magnitude greater than that of mollusk hemocyanin. The higher catalase activity of tyrosinase could be attributed to easier accessibility of H(2)O(2) to the dinuclear copper site of tyrosinase. Mushroom tyrosinase has also been demonstrated for the first time to catalyze oxygenation reaction of phenols with H(2)O(2) (peroxygenase activity). The reaction has been investigated kinetically by monitoring the H(2)O(2) consumption rate in 0.5 M borate buffer (pH 7.0) under aerobic conditions. Similarity of the substituent effects of a series of p-substituted phenols in the peroxygenase reaction with H(2)O(2) to those in the phenolase reaction with O(2) as well as the absence of kinetic deuterium isotope effect with a perdeuterated substrate (p-Cl-C(6)D(4)OH vs p-Cl-C(6)H(4)OH) clearly demonstrated that the oxygenation mechanisms of phenols in both systems are the same, that is, the electrophilic aromatic substitution reaction by a (micro-eta(2):eta(2) peroxo)dicopper(II) intermediate of oxy-tyrosinase. PMID- 15350141 TI - Expression of a nonpolymerizable actin mutant in Sf9 cells. AB - We have succeeded in expressing actin in the baculovirus/Sf9 cell system in high yield. The wild-type (WT) actin is functionally indistinguishable from tissue purified actin in its ability to activate ATPase activity and to support movement in an in vitro motility assay. Having achieved this feat, we used a mutational strategy to express a monomeric actin that is incapable of polymerization. Native actin requires actin binding proteins or chemical modification to maintain it in a monomeric state. The mutant actin sediments in the analytical ultracentrifuge as a homogeneous monomeric species of 3.2 S in 100 mM KCl and 2 mM MgCl(2), conditions that cause WT actin to polymerize. The two point mutations that render actin nonpolymerizable are in subdomain 4 (A204E/P243K; "AP-actin"), distant from the myosin binding site. AP-actin binds to skeletal myosin subfragment 1 (S1) and forms a homogeneous complex as demonstrated by analytical ultracentrifugation. The ATPase activity of a cross-linked AP-actin.S1 complex is higher than that of S1 alone, although less than that supported by filamentous actin (F-actin). AP Actin is an excellent candidate for structural studies of complexes of actin with motor proteins and other actin-binding proteins. PMID- 15350142 TI - The mechanism of beta-hairpin formation. AB - Beta-hairpins constitute an important class of connecting protein secondary structures. Several groups have postulated that such structures form early in the folding process and serve to nucleate the formation of extended beta-sheet structures. Despite the importance of beta-hairpins in protein folding, little is known about the mechanism of formation of these structures. While it is well established that there is a complex interplay between the stability of a beta hairpin and loop conformational propensity, loop length, and the formation of stabilizing cross-strand interactions (H-bonds and hydrophobic interactions), the influence of these factors on the folding rate is poorly understood. Peptide models provide a simple framework for exploring the molecular details of the formation of beta-hairpin structures. We have explored the fundamental processes of folding in two linear peptides that form beta-hairpin structures, having a stabilizing hydrophobic cluster connected by loops of differing lengths. This approach allows us to evaluate existing models of the mechanism of beta-hairpin formation. We find a substantial acceleration of the folding rate when the connecting loop is made shorter (i.e., the hydrophobic cluster is moved closer to the turn). Analysis of the folding kinetics of these two peptides reveals that this acceleration is a direct consequence of the reduced entropic cost of the smaller loop search. PMID- 15350143 TI - Assembly and stability of nisin-lipid II pores. AB - The peptide antibiotic nisin was the first reported example of an antibiotic that kills bacteria via targeted pore formation. The specific target of nisin is Lipid II, an essential intermediate in the bacterial cell-wall synthesis. High-affinity binding of the antibiotic to Lipid II is followed by rapid permeabilization of the membrane. Here, we investigated the assembly and stability of nisin-Lipid II pore complexes by means of pyrene fluorescence and circular dichroism. We demonstrated that nisin uses all available Lipid II molecules in the membrane to form pore complexes. The pore complexes have a uniform structure and consist of 8 nisin and 4 Lipid II molecules. Moreover, the pores displayed a remarkable stability, because they were able to resist the solubilization of the membrane environment by mild detergents. Similar experiments with [N20P/M21P]nisin showed that the hinge region is essential for the assembly into stable pore complexes. The new insights were used to propose a refined model for nisin pore formation. PMID- 15350144 TI - Photochromic biliproteins from the cyanobacterium Anabaena sp. PCC 7120: lyase activities, chromophore exchange, and photochromism in phytochrome AphA. AB - Photochromic biliproteins can be switched by light between two states, initiated by Z/E photoisomerization of the linear tetrapyrrole chromophore. The cyanobacterium Anabaena sp. PCC 7120 contains three genes coding for such biliproteins, two coding for phytochromes (aphA/B) and one for the alpha subunit of phycoerythrocyanin (pecA). (a) aphA was overexpressed in Escherichia coli with N-terminal His and S tags, and the protein was reconstituted by an optimized protocol with phycocyanobilin (PCB), to yield the photochromic chromoprotein, PCB AphA, carrying the PCB chromophore. (b) AphA chromophorylation is autocatalytic such as in other phytochromes. (c) AphA chromophorylation is also possible by chromophore transfer from the PCB-carrying biliprotein, phycocyanin (CPC). The autocatalytic transfer is very slow, and it is enhanced more than 100-fold by catalysis of PCB:CpcA lyase and alpha-CPC as donor. (d) Through deletion mutations of aphA, a short sequence IQPHGV [amino acids (aa) 26-31] was found essential for the lyase activity of AphA, indicating an interaction of the N terminus with the chromophore-binding domain around cysteine 259. (e) A motif of at least 23 aa, starting with this sequence and located approximately 250 aa N terminal of the chromophore-binding cysteine, is proposed to relate to the lyase function in plant and most prokaryotic phytochromes. (f) Long-range interactions in AphA are further supported by blue-shifted absorptions (50% of patients. The incidence of preventable ADRs is similar to that found in adult literature. No single drug caused >5% of reported ADRs. Opioids, anticonvulsants and antibiotics were the most common drug classes associated with ADRs. Thus, strategies targeting these drug classes and interventions during the medication ordering and administration processes may reduce the number of ADRs and possibly the associated costs. Even though preventable ADRs may not be entirely eliminated, the goal should be to increase ADR awareness and encourage early detection and intervention to minimise patient discomfort. PMID- 15350152 TI - A benefit-risk assessment of baclofen in severe spinal spasticity. AB - Baclofen is used for treatment of the spasticity of spinal origin that is a common sequela of spinal cord injury and multiple sclerosis; spasticity occurs in about 50% of patients affected by these disorders. In open-label studies of oral baclofen, the drug improved spasticity in 70-87% of patients; additionally, improvement in spasms was reported in 75-96% of patients. In double-blind, crossover, placebo-controlled trials, baclofen was reported to be effective, producing statistically significant improvements in spasticity. Tizanidine is the antispasticity drug that has been most widely compared with oral baclofen; studies have generally found the two drugs to have equivalent efficacy. However, tizanidine has better tolerability, in particular weakness was reported to be occur less frequently with tizanidine than with baclofen. The main adverse effects of oral baclofen include: sedation or somnolence, excessive weakness, vertigo and psychological disturbances. The incidence of adverse effects is reported to range from 10% to 75%. The majority of adverse effects are not severe; most are dose related, transient and/or reversible. The main risks of oral baclofen administration are related to withdrawal: seizures, psychic symptoms and hyperthermia can occur. These symptoms improve after the reintroduction of baclofen, usually without sequelae. When not related to withdrawal; these symptoms mainly present in patients with brain damage and in the elderly. The limited data on baclofen toxicity in patients with renal disease suggest that administration of the drug in these persons may carry an unnecessarily high risk. Intrathecal baclofen is indicated for use in patients with spasticity of spinal origin unresponsive to treatment with maximum doses of oral baclofen, tizanidine and/or dantrolene. The benefits of continuous intrathecal baclofen infusion have been demonstrated: >80% and >65% of patients have improvement in tone and spasms, respectively. The main risks of intrathecal baclofen infusion are symptoms related to overdose or withdrawal; the latter is more important because of the associated severe effects on clinical status and the possibility of death, but it is responsive to rapid treatment. Overdose primarily arises from drug test doses or human error during refill and programming of the pump, and withdrawal most commonly occurs as a result of a problem with the delivery system. Since the adverse consequences do not exceed the benefits of oral and intrathecal baclofen for patients with spinal spasticity, the benefit/risk assessment is favourable. PMID- 15350155 TI - Comparative tolerability of drug treatment for nocturnal enuresis in children. AB - Primary nocturnal enuresis is one of the most frequent complaints in paediatric and urologic practice. Physicians face the dilemma of whether or not to treat primary nocturnal enuresis since the trend towards spontaneous remission is countered by social disadvantages and reduced self esteem of the children affected and their families. We reviewed randomised, controlled trials investigating efficacy and adverse effects of current medical treatment for primary nocturnal enuresis. Only desmopressin and imipramine displayed significant effects in reducing wet nights: when compared with baseline bedwetting or placebo controls, 30-70% of the studied children achieved therapeutic success. For drugs such as indometacin or oxybutynin, convincing studies displaying a significant positive effect are still needed. However, considering the adverse effects profiles of desmopressin and imipramine it can be seen that imipramine is associated with about twice as many unwanted reactions. More importantly, a serious adverse effect of imipramine is sudden cardiac arrest. In general, adverse effects with desmopressin are rare and mild, but there have been a number of case reports of hyponatraemic hypervolaemia associated with coma and seizures. Of these, many cases were attributed to excess water intake before taking the drug and all children recovered fully. In summary, if medical treatment is considered, preference should be given to desmopressin. PMID- 15350154 TI - A benefit-risk assessment of inhaled long-acting beta2-agonists in the management of obstructive pulmonary disease. AB - The two inhaled long-acting beta2-adrenoceptor agonists, salmeterol and formoterol, have been studied extensively since their introduction in the early 1990s. In this review we consider the evidence for their efficacy and safety in adults with asthma and chronic obstructive pulmonary disease (COPD), by reviewing long-term prospective studies in which these drugs have been compared with placebo or an alternative bronchodilator. We have also assessed safety, including data from postmarketing surveillance studies and case-control studies using large databases. In patients with asthma, salmeterol and formoterol increase lung function, reduce asthmatic symptoms and improve quality of life when compared with placebo. Both drugs protect against exercise-induced asthma, although some tolerance develops with regular use. Tolerance to the bronchodilator effects of formoterol has also been seen, although this is small and most of the beneficial effects are maintained long-term. Both drugs have been shown to reduce asthma exacerbations but only in studies in which most patients were taking an inhaled corticosteroid. Adding a long-acting beta2-agonist provided better control than increasing the dose of inhaled corticosteroid in several studies. Long-acting beta2-agonists also provide better asthma control than use of regular short acting beta2-agonists and theophylline. Their relative efficacy compared with leukotriene antagonists is uncertain as yet. Formoterol appears to be at least as safe and effective as a short-acting beta2-agonist when used on an 'as required' basis. In patients with COPD, both salmeterol and formoterol offer improved lung function and reduced COPD symptoms compared with placebo, and quality of life has been improved in some studies. Some tolerance to the bronchodilating effect of salmeterol was seen in one study. Most studies have not found a significant reduction in exacerbations in COPD. Both drugs have provided greater benefit than ipratropium bromide or theophylline; there are limited data on tiotropium bromide. The long-acting beta2-agonists cause predictable adverse effects including headache, tremor, palpitations, muscle cramps and a fall in serum potassium concentration. Salmeterol can also cause paradoxical bronchospasm. There is some evidence that serious adverse events including dysrhythmias and life-threatening asthma episodes can occur; however, the incidence of such events is very low but may be increased in patients not taking an inhaled corticosteroid. Salmeterol 50 microg twice daily and formoterol 12 microg twice daily are effective and safe in treating patients with asthma and COPD. Higher doses cause more adverse effects, although serious adverse events are rare. PMID- 15350156 TI - Calculation of doses of drugs in solution: are medical students confused by different means of expressing drug concentrations? AB - INTRODUCTION: Our hypothesis was that clinical medical students find the different means of expressing the concentration of drugs in solution confusing. We are concerned that lack of formal teaching on this topic may make students liable to make drug dosing errors after they have qualified. Administering the wrong volume of a drug may have serious consequences for patient safety. STUDY DESIGN AND PARTICIPANT GROUP: Web-based electronic multiple-choice examination of clinical medical students. METHODS: We asked clinical medical students at our university three multiple-choice questions concerning the concentration of lidocaine (lignocaine) and epinephrine (adrenaline) in solution and the maximal recommended dose of lidocaine. The incorrect options were wrong by factors of between 4 and 1000. RESULTS: One hundred and sixty-eight clinical students out of 350 contacted responded to an invitation to participate (response rate 48%). Twenty-seven percent answered every question incorrectly and 10% answered all three correctly. The mean score for all students was only 1.24 out of 3 (standard error 0.96). However, final-year students performed significantly better (p = 0.016), implying that some knowledge had been acquired informally. Their higher mean score resulted from correctly identifying the amount of epinephrine (p = 0.005) and lidocaine (p = 0.018) more frequently. Only 27% knew the maximal recommended dose of lidocaine, with no difference between years (p = 0.724). CONCLUSIONS: A substantial majority of medical students are unable to calculate the mass of a drug in solution correctly. There is evidence that some students are picking up this skill during the course, because final-year students performed significantly better than first-year students. Modern medical student pharmacology teaching is highly sophisticated, encompassing genomics, molecular and cell biology. The ability to calculate drug doses safely appears to have been overlooked. Students should be familiar with these concepts, so as to avoid dose errors and associated morbidity, mortality and cost when they begin prescribing. To simplify calculations, drug packaging should express the concentration of drugs in solution solely as mass per unit volume, e.g. milligrams per millilitre. PMID- 15350157 TI - Safety related drug-labelling changes: findings from two data mining algorithms. AB - INTRODUCTION: With increasing volumes of postmarketing safety surveillance data, data mining algorithms (DMAs) have been developed to search large spontaneous reporting system (SRS) databases for disproportional statistical dependencies between drugs and events. A crucial question is the proper deployment of such techniques within the universe of methods historically used for signal detection. One question of interest is comparative performance of algorithms based on simple forms of disproportionality analysis versus those incorporating Bayesian modelling. A potential benefit of Bayesian methods is a reduced volume of signals, including false-positive signals. OBJECTIVE: To compare performance of two well described DMAs (proportional reporting ratios [PRRs] and an empirical Bayesian algorithm known as multi-item gamma Poisson shrinker [MGPS]) using commonly recommended thresholds on a diverse data set of adverse events that triggered drug labelling changes. METHODS: PRRs and MGPS were retrospectively applied to a diverse sample of drug-event combinations (DECs) identified on a government Internet site for a 7-month period. Metrics for this comparative analysis included the number and proportion of these DECs that generated signals of disproportionate reporting with PRRs, MGPS, both or neither method, differential timing of signal generation between the two methods, and clinical nature of events that generated signals with only one, both or neither method. RESULTS: There were 136 relevant DECs that triggered safety-related labelling changes for 39 drugs during a 7-month period. PRRs generated a signal of disproportionate reporting with almost twice as many DECs as MGPS (77 vs 40). No DECs were flagged by MGPS only. PRRs highlighted DECs in advance of MGPS (1-15 years) and a label change (1-30 years). For 59 DECs, there was no signal with either DMA. DECs generating signals of disproportionate reporting with only PRRs were both medically serious and non-serious. DISCUSSION/CONCLUSION: In most instances in which a DEC generated a signal of disproportionate reporting with both DMAs (almost twice as many with PRRs), the signal was generated using PRRs in advance of MGPS. No medically important events were signalled only by MGPS. It is likely that the incremental utility of DMAs are highly situation-dependent. It is clear, however, that the volume of signals generated by itself is an inadequate criterion for comparison and that clinical nature of signalled events and differential timing of signals needs to be considered. Accepting commonly recommended threshold criteria for DMAs examined in this study as universal benchmarks for signal detection is not justified. PMID- 15350158 TI - Haematological effects of interferon-beta-1a (Rebif) therapy in multiple sclerosis. AB - INTRODUCTION: Interferon-beta-1a (Rebif) is an established treatment for relapsing-remitting multiple sclerosis (MS) and haematological changes are commonly reported in clinical trials of this agent. The combined clinical trial and postmarketing safety database for subcutaneous interferon-beta-1a (Rebif) allows a comprehensive, retrospective assessment of both common and infrequent haematological effects associated with interferon-beta therapy. METHODS: Haematological laboratory abnormalities were analysed from six randomised, controlled clinical trials of subcutaneous interferon-beta-1a in MS, five of which were placebo-controlled. Treatment data were collected from 2482 patients for up to 6 months, 1178 patients for up to 2 years and 786 patients for up to 6 years. Total interferon-beta-1a doses ranged from 22 microg once weekly to 44 microg three times weekly. Postmarketing surveillance data were also analysed. RESULTS: Treatment with interferon-beta-1a led to asymptomatic dose-related reductions in all cell lineages under investigation, predominantly white blood cells. The greatest differences between interferon-beta-1a therapy and placebo were seen for total leucocyte and neutrophil counts. At least two-thirds of patients affected by cytopenia experienced the onset of cytopenia within the first 6 months of therapy. The majority of events were mild and generally resolved within 3--4 months, while continuing therapy. Dose reductions were uncommon and only a small proportion (6 of 727; 0.8%) of patients stopped treatment over 2 years because of haematological abnormalities when receiving the highest dose of interferon-beta-1a, 44 microg three times weekly. Postmarketing safety reports were similarly related to asymptomatic cytopenias, although one case of potentially related autoimmune haemolytic anaemia was reported. CONCLUSION: Although haematological abnormalities are common and dose-related in patients with MS receiving interferon-beta-1a, the events are mainly mild and transient, with little impact on adherence to therapy. Haematological events are rarely of clinical significance and do not adversely affect the benefit-to-risk ratio that favours high-dose interferon-beta-1a therapy. PMID- 15350159 TI - Targeting tachykinins for the treatment of obstructive airways disease. AB - The tachykinin family of peptides are distributed throughout the nervous system and are thought to play a critical role in inflammation and immunomodulation. Tachykinins have been implicated in the pathogenesis of many diseases and disease processes including inflammatory pain, emesis, depression, Parkinson's disease and inflammatory bowel syndrome. In the airways of animals, substance P and neurokinin A are released from a subset of airway sensory nerves, and evoke vasodilatation, bronchoconstriction, mucus secretion, leukocyte recruitment, airways hyperreactivity and cough. These observations have led to suggestions that tachykinins may also be viable targets for the treatment of obstructive airways disease. Clinical trials in humans assessing the utility of tachykinin receptor antagonists such as nepadutant and saredutant for the treatment of asthma are limited, and the results for the most part have been inconclusive. Several new tachykinin receptor antagonists have been recently designed to target multiple tachykinin receptor subtypes and to readily penetrate into the central nervous system. Future clinical trials with these compounds should help to shed some light on the role of tachykinins in obstructive airways disease. PMID- 15350160 TI - Pulmonary alveolar proteinosis. Clinical manifestations and optimal treatment strategies. AB - Pulmonary alveolar proteinosis (PAP) is characterized by the accumulation of surfactant phospholipids and proteins within the lung alveoli. Important advances have been made over the past 8 years in our understanding of this disease, offering new directions for research and patient care. First, genetically altered mice that are homozygous for a disrupted granulocyte-macrophage colony stimulating factor (GM-CSF) gene developed a lung lesion with histologic resemblance to PAP. The surfactant is thought to be catabolized or cleared mostly by alveolar macrophages, this process being dependent on GM-CSF. Second, a neutralizing autoantibody against GM-CSF was found in serum and bronchoalveolar lavage fluid of patients with idiopathic PAP but not in healthy controls, thereby raising the suspicion that human PAP may be an autoimmune disease. The relationship between the antibody and disease pathogenesis remains unclear but data suggest that the GM-CSF antibody may have a potential role as a diagnostic test. No specific therapy exists for PAP. Sequential whole lung lavage is the standard of care. Exogenous therapy with GM-CSF may improve the lung disease in some patients with PAP but this therapy is still experimental. Interventions directed at treating a relative GM-CSF deficiency by administration of GM-CSF or lowering the antibody level (i.e. by plasmapheresis or immunosuppression) may hold promise as future therapy for this rare disease. PMID- 15350161 TI - Pulmonary alveolar proteinosis. What is the role of GM-CSF in disease pathogenesis and treatment? PMID- 15350162 TI - Current and emerging nonsteroidal anti-inflammatory therapies targeting specific mechanisms in asthma and allergy. AB - Today inhaled corticosteroids (ICS) are regarded as the first-line controller anti-inflammatory treatment in the management of asthma. However, there is an increasing awareness of the risk of long-term adverse effects of ICS and that asthma is not only an organ-specific disease but also a systemic and small airway disease. This thinking has called for systemic treatment alternatives to treat asthma targeting more disease-specific mechanisms without influencing normal physiologic functions. Blocking of disease-specific mediators is a mechanism utilized by anti-leukotrienes and anti-immunoglobulin E treatment, each proven to be effective in both asthma and allergic rhinitis.Different cytokine-modifying strategies have been tested in clinical trials with variable results, some disappointing and some encouraging. Anti-interleukin (IL)-5 monoclonal antibody treatment effectively reduces the number of eosinophils locally in the airways and in peripheral blood in asthmatic patients. Unfortunately, this marked effect on eosinophils was not associated with an improvement in bronchial hyperresponsiveness and/or symptoms. Clinical trials with a recombinant soluble IL-4 receptor have been somewhat more successful at improving asthma control and allowing reduction of ICS therapy in asthma. Treatment with recombinant IL-12 had an effect on bronchial hyperresponsiveness and eosinophilic response, but was associated with unacceptable adverse effects. Other interesting cytokine modulating treatments include those targeting IL-9, IL-10, IL-12 and IL-13.Immune modulating treatment with bacterial antigens represents another strategy, originating from the hypothesis that some bacterial infections guide the immune system towards a T helper (Th) type 1 immune response. Mycobacterium vaccae, Bacille Calmette-Guerin (BCG) and immunostimulatory DNA sequences have all been tested in clinical trials, with encouraging results. Future asthma and allergy treatment will probably include not only one but also two or more disease modifying agents administered to the same patient. PMID- 15350163 TI - Tiotropium bromide. A review of its use as maintenance therapy in patients with COPD. AB - Tiotropium bromide (Spiriva) is a long-acting anticholinergic bronchodilator that maintains bronchodilation for at least 24 hours, allowing once-daily administration. The active moiety is the tiotropium cation (tiotropium); tiotropium bromide 22.5 micrograms is equivalent to 18 micrograms of tiotropium cation. Greater improvements in lung function from baseline (primary endpoint mean trough FEV(1)) were observed with inhaled tiotropium 18 micrograms once daily than with placebo in 6-month and 1-year randomized, double-blind trials in patients with COPD. Tiotropium improved lung function (trough FEV(1) response) more effectively than ipratropium bromide (ipratropium) 40 micrograms four times daily in 1-year clinical trials, and was at least as effective as salmeterol 50 micrograms 12-hourly in 6-month trials. Preliminary data suggest that tiotropium alone or in combination with once-daily formoterol has a greater bronchodilator effect than twice-daily formoterol in patients with COPD. Improvements in patients' perception of health-related quality of life (HR-QOL) or dyspnea were greater with tiotropium than with placebo or ipratropium, and were similar to those with salmeterol. Reductions in the frequency and severity of acute exacerbations and in the use of rescue medication were also greater with tiotropium than with ipratropium or placebo. There was no evidence of tachyphylaxis with tiotropium during 1-year clinical trials. Inhaled tiotropium was generally well tolerated in clinical trials. Apart from dry mouth, the type and incidence of adverse events with tiotropium were similar to those with ipratropium, salmeterol or placebo in patients with COPD. In conclusion, inhaled tiotropium 18 micrograms once daily improved lung function, dyspnea, and HR-QOL, and decreased the incidence of acute COPD exacerbations and the use of rescue medication relative to placebo or ipratropium in clinical trials in patients with COPD. Tiotropium was at least as effective as salmeterol in terms of bronchodilator efficacy and improvements in dyspnea or HR-QOL. With the exception of dry mouth, the tolerability profile of tiotropium was similar to that with placebo, ipratropium, or salmeterol. Consequently, inhaled tiotropium is likely to be a valuable option for first-line, long-term maintenance therapy in the management of bronchoconstriction in patients with symptomatic COPD. Tiotropium bromide has a quaternary ammonium structure and acts as an anticholinergic bronchodilator; the active moiety is the tiotropium cation (tiotropium). A 22.5 micrograms dose of tiotropium bromide provides 18 micrograms of tiotropium. Orally inhaled tiotropium bromide antagonizes the muscarinic M(1), M(2), and M(3) receptors located in airway smooth muscle, reversing vagally mediated bronchoconstriction. Receptor binding assays and in vitro tests indicate that tiotropium bromide is kinetically selective for M(1) and M(3) receptors over the M(2) receptor, unlike ipratropium bromide, which is nonselective. Animal and in vitro studies showed that tiotropium bromide was more potent ( approximate, equals 20-fold) than ipratropium bromide in displacing [(3)H]N-methylscopolamine (NMS) from muscarinic receptors, and had a more sustained protective effect (>70% inhibition) against NMS binding. Tiotropium bromide was a more potent inhibitor of bronchial contraction than atropine ( approximate, equals 23-fold), and had a slower onset and markedly longer duration of action than atropine or an equipotent dose of ipratropium bromide. Aerosol particle penetration is improved with tiotropium, without delaying mucus clearance from the lungs. Tiotropium 4.5 36 micrograms once daily for 4 weeks increased mean trough and average FEV(1) and FVC and mean PEFR values from baseline compared with placebo, with no evidence of tachyphylaxis. Improvements in trough FEV(1) from baseline with tiotropium 4.5-36 micrograms were not dose dependent. Based on a lack of dose response, the optimal once-daily tiotropium dosage is 18 micrograms. Steady-state trough FEV(1) values are achieved within 48 hours of commencing tiotrochodilation (for >/=24 hours) and an attenuation of the nocturnal decline in FEV(1) that were unaffected by timing of the daily tiotropium dose were seen in randomized, double-blind, placebo-controlled studies in patients with stable COPD. The drug improved static and dynamic lung hyperinflation (evidenced by reduced trapped air volume and increased tidal volume and end-of-exercise inspiratory capacity), and improved exertional dyspnea (during activities of daily living and exertion) and exercise tolerance compared with placebo in randomized, double-blind studies. In patients with stable COPD, improved sleep-related oxygen desaturation that was unaffected by the timing of the daily dose was seen with tiotropium but not with placebo. Clinically significant treatment-related disorders of conduction or rhythm, or changes in heart rate were not observed with tiotropium in this patient group. Mean maximal plasma concentrations (C(max)) were observed within 5 minutes of inhalation of a single dose of tiotropium 18 micrograms in patients with COPD. Plasma drug levels declined to minimum concentrations (C(min)) within 1 hour of treatment in healthy volunteers. Mean steady-state C(max) concentrations (16 ng/L) were achieved after 2-3 weeks of once-daily inhaled tiotropium 18 micrograms in elderly patients with COPD; tiotropium does not appear to accumulate once steady-state has been achieved.The estimated absolute bioavailability of tiotropium at steady state in healthy volunteers was approximately 20-25%, and approximately 72% of the drug is bound to plasma proteins. Excretion of tiotropium is predominantly renal (through active secretion by the kidneys), although in vitro studies suggest that cytochrome P450 (CYP) oxidation (possibly involving CYP2D6 and CYP3A4 enzymes) may have a minor role. In patients with COPD, renal excretion of the unchanged drug at 24 hours (Ae(24)) was approximately 7%. The mean plasma elimination half-life after single or multiple doses in healthy volunteers and elderly patients with COPD was approximately 5-6 days. The renal clearance and urinary excretion of tiotropium decrease with increasing age; however, these changes are not considered to be clinically significant. Because of altered steady-state C(max), C(min), area under the concentration-time curve, and Ae(24) values, caution is required with tiotropium administration in patients with moderate-to-severe renal impairment. The pharmacokinetics of tiotropium in patients with severe renal or hepatic impairment have not been studied. Tiotropium does not interact with drugs such as cimetidine or ranitidine, which are also eliminated by active renal secretion. Orally inhaled tiotropium bromide has been evaluated as a bronchodilator for the management of patients with COPD in randomized, double-blind 6-month and 1-year trials, and in several shorter studies. In clinical trials, COPD was diagnosed according to the American Thoracic Society guidelines. The bronchodilator effect was expressed as the trough FEV(1) response (the mean change in FEV(1) from baseline measured 1 hour prior to and immediately before a scheduled dose), and was the primary endpoint in all but two clinical trials. The bronchodilator effect with tiotropium 18 micrograms once daily was superior to that with placebo in several well designed trials in patients with COPD. Moreover, greater improvements in mean peak and average FEV(1) responses occurred with tiotropium but not with placebo. Mean trough, peak, and average FVC responses, and weekly mean morning and evening PEFR values were also improved to a greater extent with tiotropium than with placebo. Tiotropium demonstrated a greater bronchodilator effect than ipratropium bromide (hereafter referred to as ipratropium when used at approved dosages) 40 micrograms four times daily in two 1-year trials in patients with COPD. Mean peak and average FEV(1), mean trough FVC responses, and weekly mean morning and evening PEFR values were also increased to a greater extent with tiotropium than with ipratropium. In one of the two 6-month trials that compared the efficacy of tiotropium with that of inhaled salmeterol 50 micrograms twice daily, greater improvements from baseline in mean trough, peak, and average FEV(1) and FVC responses were seen with tiotropium than with salmeterol. Increases in weekly mean evening, but not morning, PEFR values were generally greater with tiotropium than salmeterol. In the second trial, improvement in the primary endpoint (mean trough FEV(1) response from baseline) with tiotropium or salmeterol was similar, although peak and average responses were superior with tiotropium. Preliminary results from a 6-week crossover study in patients with COPD suggested that tiotropium alone or in combination with once daily formoterol improved mean trough and average FEV(1) and trough FVC values from baseline to a greater extent than twice-daily formoterol. More patients achieved a clinically important improvement (increase of >/=1 unit) in the transitional dyspnea index focal score (a measure of dyspnea-related impairment) with tiotropium than with placebo in the 1-year trials. Tiotropium was superior to ipratropium in 1-year trials, and was at least as effective as salmeterol in 6 month trials, in achieving a clinically important improvement in focal scores. Tiotropium recipients experienced fewer COPD exacerbations than placebo or ipratropium recipients and had fewer and shorter COPD-related hospitalizations compared with placebo recipients. Unlike salmeterol, tiotropium lengthened the time to onset of the first exacerbation and decreased the number of exacerbations compared with placebo in two 6-month trials. Similar proportions of tiotropium, salmeterol, and placebo recipients required COPD-related hospitalizations. (ABSTRACT TRUNCATED) PMID- 15350164 TI - A new era in atherosclerosis drug discovery. PMID- 15350166 TI - Fluvastatin and fluvastatin extended release: a clinical and safety profile. AB - Cardiovascular diseases due to atherosclerosis are the leading causes of mortality in the Western world. Cholesterol-lowering therapy with 3-hydroxy-3 methylglutaryl coenzyme Areductase inhibitors (statins) has demonstrated a reduction in cardiovascular morbidity and mortality in diverse populations. Fluvastatin (Lescol, Novartis Pharmaceuticals) was the first totally synthetic 3 hydroxy-3-methylglutaryl coenzyme A reductase inhibitor on the market and has recently become available in an extended-release formulation (Lescol XL, Novartis Pharmaceuticals). Data from several clinical outcome trials have shown substantial benefits from fluvastatin treatment in diverse populations. Fluvastatin exists primarily in its acid form and as inactive metabolites in vivo, while active metabolites as well as the lactone form are only present in small amounts. The demonstration of the safe use of fluvastatin in a wide range of patients may be associated with the predominant acid form of the drug in vivo, as well as its predominant metabolism via the cytochrome P450 2C9 pathway. PMID- 15350167 TI - Dexamethasone-eluting stent: an anti-inflammatory approach to inhibit coronary restenosis. AB - The long-term efficacy of percutaneous coronary interventions is still hampered by restenosis. Restenosis is the result of a complex pathophysiological process, which is thought to be caused by an exaggerated healing response induced by the vascular injury caused by the percutaneous coronary interventions and the implantation of a foreign body (the stent). There is increasing evidence that inflammation plays an important role in the initiation and development of neointimal hyperplasia and subsequent restenosis. Dexamethasone (Decadron, Merck Sharpe and Dohme Ltd) is a glucocorticoid with well-known potent anti inflammatory and antiproliferative properties. Early studies using either systemic or local delivery of dexamethasone have shown limited beneficial effects on restenosis. The dexamethasone-eluting stent (Dexamet, Abbott Vascular Devices Ltd) is one of the first generation of drug-eluting stents for local drug delivery to prevent restenosis. Preclinical studies demonstrated that implantation of dexamethasone-loaded coronary stents was safe and had a beneficial effect on stent implantation-related inflammation. A pilot trial suggested a beneficial effect on restenosis. Large randomized trials are underway to confirm these findings. This article reviews the potential role of inflammation in the pathogenesis of restenosis and the efficacy of dexamethasone in the prevention of restenosis. PMID- 15350168 TI - Recombinant Factor VIIa: clinical applications for an intravenous hemostatic agent with broad-spectrum potential. AB - Recombinant Factor VIIa (NovoSeven, Novo Nordisk) is a unique hemostatic agent with potential for broad-spectrum applications in bleeding patients with congenital and acquired bleeding abnormalities. At present, recombinant Factor VIIa is only approved for the treatment of hemophilia A and B patients who have acquired antibodies to Factors VIII and IX. However, the literature is rapidly expanding indicating that rFVIIa could benefit patients with bleeding due to a variety of etiologies. Unfortunately, the vast majority of these reports are case studies or small-series summaries, and are neither prospective nor controlled. Controlled trials are currently underway in several potential areas of application. While the possibility of thrombotic complications has been a clinical concern, safety data on patients treated to date have not revealed a significant problem. Expansion of the clinical applications for this interesting and important hemostatic agent are hampered by its expense, the limited scope of the US Food and Drug Administration approval for its use, and the absence of an appropriate laboratory monitoring assay. PMID- 15350169 TI - Amlodipine/atorvastatin: the first cross risk factor polypill for the prevention and treatment of cardiovascular disease. AB - In 2002, the World Health Organization estimated that over 58% of cardiovascular disease in North America is due to 'both blood pressure and cholesterol higher than optimal'. Unfortunately, less than a third of patients with both conditions are identified, and fewer than one in ten reach the treatment goals for both factors. Adherence to treatment is notably improved when therapy is initiated simultaneously. Combination therapy of amlodipine besylate (Norvasc, Pfizer Ltd) with atorvastatin calcium (Lipitor, Pfizer Ltd), marketed as Caduet (Pfizer Ltd) is the first dual-therapy compound designed to treat hypertension and/or angina and dyslipidemia concurrently with a single daily pill in the full range of dosing combinations. Amlodipine/atorvastatin retains the safety and efficacy of its parent compounds whilst simplifying the management of these comorbid conditions, in what may be considered the first version of a polypill. PMID- 15350170 TI - Genetics of inherited cardiomyopathies. AB - Cardiomyopathies are the most common disorders resulting in heart failure, with dilated cardiomyopathy being responsible for the majority of cases. Other forms of cardiomyopathy, especially hypertrophic forms, are also important causes of heart failure. The mortality rate due to cardiomyopathy in the USA is over 10,000 deaths per year, and the costs associated with heart failure are approximately 200 million US dollars per year in the USA alone. Over the past few years, breakthroughs have occurred in understanding the basic mechanisms of these disorders, potentially enabling clinicians to devise improved diagnostic strategies and therapies. As at least 30 to 40% of cases are inherited, it is now imperative that the genetic basis for these disorders is clearly recognized by caregivers and scientists. However, it has also become clear that these diseases are genetically highly heterogeneous, with multiple genes identified for each of the major forms of cardiomyopathy, and most patients having private mutations. These data suggest that the genetic diagnosis of most patients with cardiomyopathy will be impractical with current technologies. However, there are a few exceptions, such as patients with X-linked cardiomyopathies, with or without the concomitant abnormalities of cyclic neutropenia and 3 methylglutaconic aciduria, or patients with cardiomyopathy associated with conduction disease: these appear to be associated with mutations in a small subset of genes, and can be investigated by certified diagnostic laboratories. This review will summarize current knowledge of the genetics of inherited cardiomyopathies and how findings from research laboratories may be translated into the diagnostic laboratory. PMID- 15350171 TI - State of the art catheter interventions in adults with congenital heart disease. AB - Catheter interventions in adults with congenital heart disease have rapidly advanced. Transcatheter valve repair and replacement techniques have been added to the existing spectrum of well-established procedures. This review summarizes current transcatheter management strategies for congenital cardiac anomalies seen in the adult population. PMID- 15350172 TI - Transcatheter closure of atrial septal defects. AB - Transcatheter closure of atrial septal defects has evolved significantly over the last 20 years. Transcatheter closure has replaced surgical closure for the treatment of most secundum atrial septal defectsat the current time. A major reason for this is the lower morbidity of transcatheter closure procedures. Several closure devices have come into clinical use. The Amplatzer septal occluder (AGA Medical Co.) currently has the largest reported experience and subsequently the best-established safety and efficacy features. Clinically challenging situations, such as larger atrial septal defects, atrial septal defects with deficient rims and multiple atrial septal defects, are increasingly being addressed using the transcatheter approach, with improved results. The incidence of most complications has significantly reduced over time, and serious side effects are relatively uncommon. In this review, the literature is summarized regarding the current role of transcatheter closure, the evolution of the different available devices for clinical use and the complications that occur with their use. A comparison is also made with surgical closure techniques. PMID- 15350173 TI - Noninvasive assessment of coronary atherosclerosis by multidetector-row computed tomography. AB - Assessment of atherosclerotic plaque burden may help to further stratify asymptomatic subjects with an intermediate cardiac event risk according to their conventional risk factors. Coronary calcium screening is a simple and effective method to noninvasively assess the atherosclerotic plaque burden. Standardized quantification of the coronary calcium mass will allow the results of ongoing prospective cohort studies to be used for any computed tomography (CT) scanner, electron-beam CT, as well as multidetector-row CT. Coronary multidetector-row CT angiography may have the potential to visualize vulnerable plaques that are prone to rupture and cause acute coronary symptoms. However, neither the reliability of detection nor the strategies for intervention of vulnerable plaques with multidetector-row CT have to date been proven. PMID- 15350174 TI - Epidemiology, pathophysiology and therapeutic implications of lipoprotein(a) in kidney disease. AB - Chronic kidney disease is associated with a tremendously increased risk for cardiovascular disease. Traditional risk factors for cardiovascular disease, however, show a diminished predictive power in these patients compared with the general population. This review provides an overview of lipoprotein(a), which is considered a nontraditional risk factor. The characteristic genetic and nongenetic changes of lipoprotein(a) in kidney disease are discussed and set into the context of risk prediction. In particular, genetically determined apolipoprotein(a) polymorphism is a powerful risk predictor for cardiovascular disease and total mortality in these patients. Finally, the limited interventional strategies available to lower lipoprotein(a) are considered. PMID- 15350175 TI - Clinical trials in restenosis with 7-hexanoyltaxol and paclitaxel-eluting stents. AB - Restenosis rates are lower with stent implantation than with balloon angioplasty. Nevertheless, even with the use of stenting, restenosis still occurs in approximately a third of patients with diabetes, small coronary vessels and long lesions. Animal studies with paclitaxel (Taxol, Bristol-Myers Squibb)-eluting stents, mainly in endothelium-denuded normal vessels, have shown that although paclitaxel reduces restenosis in the short-term, this may only delay rather than prevent restenosis. In clinical trials, stents eluting the paclitaxel derivative, 7-hexanoyltaxol, or paclitaxel without a polymer, also delay rather than prevent restenosis. Slowing the release of paclitaxel with a polymer base in the TAXUS trade mark series of clinical trials reduced the revascularization rate at 12 months, indicating that polymer-based paclitaxel is effective for longer periods of time. Further follow-up is necessary to determine whether polymer-based release of paclitaxel represents a longer delay or prevention of restenosis. To date, paclitaxel is showing promise as an eluting agent to prevent restenosis associated with stenting. PMID- 15350176 TI - Breast arterial calcifications as a cardiovascular risk marker in women. AB - Cardiovascular disease and breast cancer are the two main causes of mortality in women. Mammography as a screening test for breast cancer is recommended for all women from the age of 40 to 50 years. Mammographically detected breast arterial calcifications are currently considered an incidental finding without clinical importance, since they are not associated with an increased risk of breast cancer. However, recent studies have shown that breast arterial calcifications on mammograms have been associated with cardiovascular risk factors and atherosclerotic cardiovascular disease. These results suggest that breast arterial calcifications, detected during routine mammography, are a noteworthy finding that could be valuable in identifying asymptomatic women at increased cardiovascular risk. PMID- 15350177 TI - Prevalence, pathophysiology and treatment of isolated systolic hypertension in the elderly. AB - Isolated systolic hypertension is the predominant type of hypertension in the elderly and is associated with cardiovascular complications such as stroke, coronary heart disease and heart failure. In this review, the role of arterial stiffness, endothelial function, atherosclerosis and oxidative stress in the pathogenesis of isolated systolic hypertension is extensively discussed. Placebo controlled intervention trials such as the Systolic Hypertension in Europe Trial and the Systolic Hypertension in the Elderly Program have clearly shown that pharmacological treatment of isolated systolic hypertension improves outcome in the elderly. Nevertheless, isolated systolic hypertension remains the major subtype of untreated and uncontrolled hypertension. PMID- 15350178 TI - Blood pressure-lowering drugs: essential therapy for some patients with normal blood pressure. AB - The risk of increasing blood pressure on the incidence of cardiovascular disease starts at 115/75 mmHg and roughly doubles for every 10 years increase in age, 20 mmHg increase in systolic blood pressure, 10 mmHg increase in diastolic blood pressure, or in the presence of comorbidities, such as diabetes or any evidence of cardiovascular disease. To lower blood pressure in patients with normal blood pressure and diabetes, or heart failure, or with any evidence of atherosclerotic disease in the coronary, cerebral and peripheral territories, reduces the incidence of major cardiovascular events by 18 to 42%. The diagnosis of hypertension in patients with these conditions is therefore irrelevant. The drugs that have mainly been tested in such conditions are the angiotensin-converting enzyme inhibitors, but their efficacy probably derives from their blood pressure lowering effect, instead of a primary antiatherosclerotic effect. PMID- 15350179 TI - Rationale for clinical trials of coagulation: reactive drugs in hepatocellular carcinoma. AB - Evidence for the regulation of cancer growth by components of the blood coagulation mechanism provides abundant opportunity for the development of novel hypotheses for the experimental treatment of malignancy. Information available on the heterogeneity in mechanisms of interaction between various cancer cell types, and procoagulant and fibrinolytic pathways, platelets, glycosaminoglycan regulated growth factors and cell-adhesion molecules indicates that insightful clinical trial design may allow targeting of individual cancer cell types with agents capable of intercepting mechanisms of growth control that are relevant to specific tumor types. This paper reviews the evidence that the common anticoagulant, heparin, inhibits hepatocellular carcinoma cell proliferation and hepatocellular carcinoma tumor dissemination in experimental animals. Clinical trials of heparin performed to date have shown increased tumor response rates and survival in other tumor types. Expression of urokinase-type plasminogen activator by hepatocellular carcinoma cells enhances tumor cell proliferation, motility, invasiveness and metastatic dissemination. Inhibition of the urokinase-type plasminogen activator/plasmin system by protease inhibitors such as aprotinin (Trasylol, Bayer) have shown improvement in the clinical course of certain tumor types. These data suggest that drugs that are well-known in the field of vascular medicine may find a role in the treatment of hepatocellular carcinoma, a common tumor type that has resisted containment by other means. PMID- 15350180 TI - Potentiation of androgen receptor transcriptional activity by inhibition of histone deacetylation--rescue of transcriptionally compromised mutants. AB - Androgens are critical in the development and maintenance of the male reproductive system and important in the progression of prostate cancer. The effects of androgens are mediated by the androgen receptor (AR), which is a ligand-modulated transcription factor that belongs to the nuclear receptor superfamily. We and others have previously shown that CREB-binding protein (CBP) can function as a coactivator for AR. Similar to some other nuclear receptor coactivators and/or the proteins that they interact with, CBP has histone acetyl transferase (HAT) activity that is thought to contribute to transcriptional activation by nuclear receptors. We have therefore assessed whether an increase in the histone acetylation status in the cell can influence AR transcriptional activity, by using the histone deacetylase (HDAC) inhibitors (HDACIs) trichostatin A (TSA), sodium butyrate (Na-But) and depsipeptide (FR901228). We found that inhibition of HDAC activity significantly increased the ability of endogenous AR in LNCaP cells, or ectopically expressed AR in HeLa cells, to activate transcription from AR-dependent reporter constructs. In addition, HDACIs increased the androgen-dependent activation of the prostate-specific antigen (PSA) gene in LNCaP cells, an increase that was not due to an increase in nuclear AR protein levels. Moreover, the viral oncoprotein E1A that inhibits CBP HAT activity fully repressed the ability of HDACIs to stimulate AR-mediated transcription, indicating that CBP is involved in this process. Deletional mutagenesis of AR indicated that whereas the AF-2 domain in the C-terminus is dispensable, the AF-1 domain in the N-terminus is required for augmentation of AR action by HDACIs, an observation which is in concordance with the reduced ability of CBP to activate AR N-terminal deletion mutants. Furthermore, HDACI treatment rescued the deficiency in the transactivation potential of AF-2 mutants. Taken together, our findings suggest that a change in the level of histone acetylation of target genes is an important determinant of AR action, possibly mediated by CBP. PMID- 15350181 TI - Serum sex hormones in men occupationally exposed to dichloro-diphenyl-trichloro ethane (DDT) as young adults. AB - To explore endocrine effects in relation to para,para'-dichloro-diphenyl-dichloro ethylene (p,p'-DDE) body burden and past occupational exposure to its precursor dichloro-diphenyl-trichloro ethane (DDT), we assayed serum sex hormones, including serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17beta-estradiol (E2), testosterone and sex hormone binding globulin (SHBG), and p,p'-DDE levels in 107 male participants in a 1946-1950 anti-malarial campaign in Sardinia, Italy. Cumulative DDT exposure during the anti-malarial operations was retrospectively estimated from detailed reports of the anti-malarial agency. Ortho,para-DDE, and its precursor ortho,para-DDT were always below the detection limit. p,p'-DDT was detected in 14/107 subjects, and p,p'-DDE in 106/107 subjects. The median lipid-adjusted p,p'-DDE serum concentration over the total study population was 396 parts per billion (interquartile range 157-1045), and it did not vary according to the job at the time of anti-malarial operations, nor was it affected by cumulative DDT exposure. LH, FSH, and SHBG, but not testosterone or E2, showed a significant positive correlation with age. Neither current serum p,p'-DDE nor past cumulative DDT exposure affected sex hormone concentrations. Our results suggest that (1) the low current p,p'-DDE serum concentration does not affect serum hormone levels, and (2) past cumulative DDT exposure is not correlated with the current p,p'-DDE serum level, nor does it show persistent effects on serum hormone levels. PMID- 15350182 TI - Raloxifene and hormone replacement therapy increase arachidonic acid and docosahexaenoic acid levels in postmenopausal women. AB - Estrogens may affect the essential n-6 and n-3 fatty acids arachidonic acid (AA; C20:4n-6) and docosahexaenoic acid (DHA; C22:6n-3). Therefore, we investigated the long-term effects of hormone replacement therapy and raloxifene, a selective estrogen-receptor modulator, in two randomized, double-blind, placebo-controlled studies. In study I, 95 healthy, non-hysterectomized, early postmenopausal women (age range 47-59 years) received one of the following treatments: daily raloxifene 60 mg (n=24), daily raloxifene 150 mg (n=23), 0.625 mg conjugated equine estrogens (CEE) plus 2.5 mg medroxyprogesterone acetate (MPA; n=24), or placebo (n=24). In study II, 30 men (age range 60-69 years) received daily 120 mg raloxifene (n=15) or placebo (n=15). In study I, plasma cholesteryl ester fatty acids were measured at baseline and after 6, 12, and 24 months in 83 (drop out rate 13%), 73 (23%), and 70 (25%) women respectively. In study II, fatty acids were measured at baseline and after 3 months in 29 men (drop out rate 3%). In postmenopausal women, administration of 150 mg raloxifene increased AA by a mean of +6.1% (P=0.055, not significant). Administration of CEE plus MPA increased AA by +14.1% (P<0.0005). Mean changes in DHA were +22.1% (P=0.003) and +14.9% (P=0.047) respectively, as compared with placebo. In men, 120 mg raloxifene for 3 months did not significantly affect AA (-5.2%; P=0.342) or DHA (+4.0%; P=0.755), but it increased testosterone levels by +19.8% (P=0.006). Administration of raloxifene 150 mg/day as well as CEE plus MPA to postmenopausal women increases the proportion of AA and DHA in plasma cholesteryl esters during a follow-up of 2 years. Short term administration of raloxifene in elderly men did not affect AA or DHA. The synthesis of AA and DHA from precursors may be enhanced through an estrogen receptor-dependent pathway. PMID- 15350183 TI - Effects of nutritional status and gonadal steroids on expression of appetite regulatory genes in the hypothalamic arcuate nucleus of sheep. AB - Sheep exhibit photoperiod-driven seasonal changes in appetite and body weight so that nutritional status increases in long days (LD) and decreases in short days (SD); additionally, they are reproductively active in SD and inactive in LD. We addressed the hypothesis that appetite-regulatory genes in the hypothalamus respond differently to changes in nutritional feedback induced by photoperiod as opposed to food restriction, and that responses would be influenced by gonadal steroid status. Castrated oestradiol-implanted male sheep were kept in SD (8 h light/day) or LD (16 h light/day) for 11 weeks, with ad libitum or restricted food (experiment 1; n=8/group). Rams were kept in SD or LD for 12 weeks with ad libitum or restricted food (experiment 2; n=6/group). Gene expression (by in situ hybridisation) in the hypothalamic arcuate nucleus for leptin receptor (OB-Rb), neuropeptide Y (NPY), pro-opiomelanocortin (POMC) and agouti-related peptide (AGRP) was unaffected by photoperiod treatment, but food restriction increased NPY and AGRP mRNAs, in experiment 1. In experiment 2, mRNAs for POMC and cocaine- and amphetamine-regulated transcript (CART) were up-regulated and AGRP down regulated in SD, while food restriction increased OB-Rb mRNA, increased NPY and AGRP mRNAs only in LD and decreased POMC mRNA only in SD. Thus, gene expression responded differently to photoperiod and food restriction, and the melanocortin pathway was up-regulated in SD in reproductively activated rams but not in oestradiol-implanted castrates. These data support the hypothesis that hypothalamic appetite-regulatory pathways respond differently to changes in nutritional feedback induced by photoperiod as opposed to food restriction, with gonadal steroid feedback additionally influencing the responses. PMID- 15350184 TI - Leptin modulates fertility under the influence of elevated growth hormone as modeled in oMt1a-oGH transgenic mice. AB - Elevated growth hormone (GH) concentrations suppress reproductive function in a variety of species, although it is unclear whether GH directly suppresses reproductive performance, or whether GH activates other pathways to achieve these effects. The ovine metallothionein 1a-ovine GH (oMt1a-oGH) transgenic mouse has been used to model the effects of GH on both body composition and reproductive function. A recent report has documented increased leptin levels in obese oMt1a oGH mice. Given the importance of leptin in modulation of the reproductive endocrine axis, as well as the reports documenting reduced leptin signal transduction in animals with elevated leptin levels, we hypothesized that high leptin concentrations in response to elevated GH would reduce fertility. To determine the effects of high circulating leptin levels on the reproductive endocrine axis, we assessed hypothalamic neuropeptide Y (NPY) and GnRH expression. At weaning, oMt1a-oGH transgenic (TG) and wild-type (WT) female mice were allocated to one of four treatment groups: oMt1a-oGH females chronically expressing the transgene (TG ON); oMt1a-oGH females expressing the transgene from 3 to 8 weeks of age (TG ON/OFF); WT females receiving the transgene stimulus from 3 to 8 weeks of age (WT ON/OFF); and WT females never receiving the transgene stimulus (WT OFF). Eight-week-old females were housed with males for a 2-week period, after which females were isolated from males and allowed to carry pregnancies to term. Body and gonadal fat pad (GFP) weights, along with plasma leptin concentrations, estrous cyclicity, pregnancy rate and litter characteristics, were recorded for each female. Chronic expression of the oMt1a oGH transgene resulted in larger leaner mice, and inactivation of the transgene produced obese females. Pregnancy rate was reduced in TG ON females when compared with all other groups, and infertility was associated with elevated leptin levels. In addition, high leptin levels were associated with increased NPY expression, suggesting reduced leptin-signaling capacity, which may contribute to suppression of the reproductive axis in oGH animals. PMID- 15350185 TI - Increased in vivo phosphorylation of insulin receptor at serine 994 in the liver of obese insulin-resistant Zucker rats. AB - Serine phosphorylation of the insulin receptor (IR) has been proposed to exert an inhibitory influence on its tyrosine kinase activity. Previous works using site directed mutagenesis suggested that serine 994 of the IR (IR Ser 994) might be part of an inhibitory domain of the receptor. In this study we examined whether this residue is subjected to phosphorylation in vivo. We used a site phosphospecific antibody to determine the extent of phosphorylation of IR Ser 994 in insulin target tissues from two animal models of insulin resistance with different IR kinase (IRK) activity: obese (fa/fa) Zucker rats and transgenic mice overexpressing bovine growth hormone (PEPCK-bGH mice). Phosphorylation at IR Ser 994 was markedly increased in liver of obese rats. This alteration appeared to be tissue-selective since no phosphorylation on Ser 994 was detected in IRs isolated from skeletal muscle of these animals. On the other hand, the phosphorylation level of IR Ser 994 was very low in liver of PEPCK-bGH mice and did not differ from that of the control group. We have also demonstrated that protein kinase (PK) C isoforms alpha, betaI and zeta are able to promote the in vitro phosphorylation of the IR on Ser 994. Differential findings in these two models of insulin resistance might thus reflect increased PKC activity resulting from increased lipid availability in obese Zucker rats. Our results suggest that Ser 994 is a novel in vivo IR phosphorylation site that might be involved in the regulation of the IRK in some states of insulin resistance. PMID- 15350186 TI - Overexpression of human IGF-II mRNA in the brain of transgenic mice modulates IGFBP-2 gene expression in the medulla oblongata. AB - The insulin-like growth factors, IGF-I and IGF-II, and their binding proteins play an important role in the growth and development of the central nervous system. In the brain, colocalization of IGFs and IGFBPs often occurs, suggesting that IGFBPs can modulate IGF action. In one strain of our human (h)IGF-II transgenic mice, which carry an hIGF-II transgene driven by the H-2Kb promoter, we found overexpression of hIGF-II in the brain, as measured by Northern blot analysis. To clarify the localization and influence of the hIGF-II transgene on different components of the GH-IGF axis in the brain, we studied the expression pattern of the hIGF-II transgene, endogenous IGF-I and IGF-II, and IGFBP-2, -3 and -5 in the brain of prepubertal 4-week-old mice, using nonradioactive in situ hybridization. We found that the hIGF-II transgene is exclusively expressed in neurons of the piriform cortex, the cerebral cortex, the medulla oblongata and the granular layer of the cerebellum. In general, this pattern is comparable to the expression pattern of endogenous IGF-I, with a few exceptions: there is no expression of IGF-I in the granular layer of the cerebellum, whereas the Purkinje cells of the cerebellum and thalamus both express IGF-I but no hIGF-II transgene. This hIGF-II transgene expression pattern contrasts markedly with endogenous IGF II expression, which is mainly located in nonneuronal cells such as the meninges and choroid plexus, and in some nuclei of the medulla oblongata. The hIGF-II transgene affects neither endogenous IGF-I and IGF-II expression, nor the expression of IGFBP-3, which is located in the choroid plexus. Although the hIGF II transgene is expressed in neuronal structures similar to IGF-I and IGFBP-5, it is not able to regulate IGFBP-5 expression, as has previously been reported for IGF-I. In the medulla oblongata, the IGFBP-2 expression level showed 10-fold upregulation by the transgene, suggesting a modulating role for IGFBP-2 at the hIGF-II transgene action in this region. PMID- 15350187 TI - Proteolytic degradation of IGF-binding protein (IGFBP)-2 in equine ovarian follicles: involvement of pregnancy-associated plasma protein-A (PAPP-A) and association with dominant but not subordinated follicles. AB - In the ovary of mammalian species, terminal follicular growth is accompanied by a decrease in intrafollicular levels of IGF-binding protein-2 (IGFBP-2) and IGFBP 4. The decrease in IGFBP-4 levels is essentially due to an increase in proteolytic cleavage by intrafollicular pregnancy-associated plasma protein-A (PAPP-A) in growing healthy follicles. The decrease in IGFBP-2 levels is partly due to a decrease in mRNA expression by follicular cells. In addition, we have recently shown that IGFBP-2 is also proteolytically cleaved by PAPP-A in bovine and porcine growing follicles. In the present work, we showed that follicular fluid from late dominant equine follicles (35 mm diameter) contains a proteolytic activity against IGFBP-2. First follicular fluid from dominant follicles contained lower levels of native IGFBP-2 than the corresponding serum, as assessed by Western ligand blotting. In contrast, immunoblotting experiments showed much higher levels of a 12 kDa proteolytic fragment in dominant follicular fluid than in the serum. Moreover, equine dominant follicular fluid was able to induce proteolysis of exogenous recombinant bovine (rb)IGFBP-2, this degradation being dose-dependently enhanced by IGFs. The proteolytic activity against IGFBP-2 in equine follicles was partially immunoneutralized by a polyclonal antibody raised against human PAPP-A. Moreover, cleavage of rbIGFBP-2 by equine follicular fluid was dose-dependently inhibited by a peptide derived from the heparin binding domain of IGFBP-5, as well as by peptides derived from other heparin binding domain-containing proteins such as connective tissue growth factor, vitronectin and heparin-interacting protein, previously shown to inhibit PAPP-A. Finally, the proteolytic activity was very low in subordinate follicles, was high in both early (25 mm diameter) and late (35 mm diameter) dominant follicles, and was slightly lower in preovulatory follicles recovered 35 h after human chorionic gonadotropin (hCG) treatment.Overall, these data show that in the equine ovary, the selection of dominant follicles is associated with an increase of the proteolytic degradation of IGFBP-2 by PAPP-A, as for IGFBP-4, and potentially other protease(s), probably contributing to the increase in IGF bioavailability. In atretic subordinate follicles, the decrease in the proteolytic degradation of IGFBP-2, probably due in part to a direct inhibition by peptides containing heparin-binding domains, contributes to the increase in IGFBP-2 levels and the decrease in IGF bioavailability. The expression of PAPP-A and IGFBP-2 mRNA during folliculogenesis remain to be investigated in the mare. PMID- 15350188 TI - Differential regulation of glucose transporter expression by estrogen and progesterone in Ishikawa endometrial cancer cells. AB - Estrogen replacement therapy and other unopposed estrogen treatments increase the incidence of endometrial abnormalities, including cancer. However, this effect is counteracted by the co-administration of progesterone. In the endometrium, glucose transporter (GLUT) expression and glucose transport are known to fluctuate throughout the menstrual cycle. Here, we determined the effect of estrogen and progesterone on the expression of GLUT1-4 and on the transport of deoxyglucose in Ishikawa endometrial cancer cells. Cells were incubated with estrogen, progesterone or combined estrogen and progesterone for 24 h and the effect on the expression of GLUT1-4 and on deoxyglucose transport was determined. We show that GLUT1 expression is upregulated by estrogen and progesterone individually, but that combined estrogen and progesterone treatment reverses this increase. Hormonal treatments do not affect GLUT2, GLUT3 or GLUT4 expression. Transport studies demonstrate that estrogen increases deoxyglucose transport at Michaelis-Menten constants (Kms) corresponding to GLUT1/4, an effect which disappears when progesterone is added concomitantly. These data demonstrate that different hormonal treatments differentially regulate GLUT expression and glucose transport in this endometrial cancer cell line. This regulation mirrors the role played by estrogen and progesterone on the incidence of cancer in this tissue and suggests that GLUT1 may be utilized by endometrial cancer cells to fuel their demand for increased energy requirement. PMID- 15350189 TI - Thyroid papillary carcinoma: preliminary evidence for a germ-line single nucleotide polymorphism in the Fas gene. AB - The expression of Fas in thyroid tumours and Graves' disease was analysed by mRNA transcript expression. As compared with unaffected thyroid tissue, Fas expression was enhanced in Graves' disease, adenomas, and papillary and follicular carcinomas. This pattern was also reflected in immunohistochemical studies. The PCR single-strand conformational polymorphism (SSCP) method and DNA sequencing were used to analyse Fas exons 1-9. The study was carried out on five different histotypes of thyroid tumours (n=93) and tissue from Graves' disease patients. As compared with a group of healthy blood donors (n=64), a significant association (P=0.006) emerged between papillary thyroid carcinoma and a silent single nucleotide polymorphism (SNP, 988C-->T) in exon 7 of the Fas gene. Other forms of thyroid pathology were not associated with the above polymorphism. Patients with neoplasia showed the same SNP in tumour tissue, in the unaffected contralateral thyroid lobe, and in peripheral blood cells. Thus, the 988C-->T polymorphism appeared to be of germ-line origin. PMID- 15350190 TI - Functional characterization of HUVEC-CS: Ca2+ signaling, ERK 1/2 activation, mitogenesis and vasodilator production. AB - While many endothelial cell lines exist, few are of human origin with characteristics close to the parent endothelial cell. We derived a subline (HUVEC CS) of immortalized human umbilical vein endothelial cells (HUVEC-C) that proliferate in standard growth media and exhibit positive acetylated low-density lipoprotein (AcLDL) uptake, express eNOS, CD31 and ve-cadherin, and spontaneously form capillary-like structures when grown on Matrigel. HUVEC-CS also maintain endothelial cell characteristics at the level of mitogenesis, kinase activation and vasodilator production. Like primary HUVEC cells, HUVEC-CS express many of the key proteins necessary for vasodilator production, including epithelial nitric oxide synthase (eNOS), HSP 90, cav-1 and -2, cPLA2, and COX-1 and -2. Prostaglandin I synthase (PGIS) was not detectable by Western blot analysis, consistent with primary HUVEC in which PGI2 production is minimal. Receptors were detected for angiotensin II (AII), bradykinin, ATP and growth factors. ATP induced a dose- and time-dependent rise in the intracellular free Ca2+ concentration ([Ca2+]i). Initially, ATP stimulates P2Y receptors rather than P2X receptors, as demonstrated by the inability of ATP to initiate a Ca2+ response subsequent to emptying of the internal Ca2+ stores by thapsigargin. AII, bradykinin, epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) also caused a rise in [Ca2+]i in a subset of the cells. ATP, basic fibroblastic growth factor (bFGF), EGF and VEGF induced mitogenesis and caused a rise in ERK 2 activation within 10 min. L-Arginine to L-citrulline conversion assays showed that ATP, EGF and VEGF induced a significant rise in eNOS activity, and this correlates with an ability to induce Ca2+ mobilization and ERK 2 activation. In conclusion, HUVEC-CS are indeed endothelial cells and appear to be functionally very similar to primary HUVEC. These cells will prove a valuable tool for future studies in both basic and therapeutic sciences. PMID- 15350191 TI - Identification of the structural and functional determinants of the extracellular domain of the human follicle stimulating hormone receptor. AB - The extracellular domain (ECD) of the human follicle-stimulating hormone receptor (hFSHR) is believed to be the major determinant of hormone selectivity. Different discrete, discontinuous regions on the ECD of the hFSHR have been suggested to be crucial for hormone binding. However, the role of the ECD in signal transduction is not well understood. This study provides some insight into these aspects of the structure-function relationship of the ECD of hFSHR. Ten peptides were selected from the ECD on the basis of their ability to be surface oriented, synthesized by the solid-phase method using fluorenylmethyloxycarbonyl chemistry, purified and characterized. They were further studied for their ability to modulate both human follicle-stimulating hormone (hFSH)-FSHR binding and cAMP generation. Competitive inhibition studies showed that, of all the peptides studied, peptides 285-300 and 297-310 hFSHR were able to inhibit hFSH binding to FSHR. Both peptides function as weak competitive inhibitors of hFSH-FSHR binding. Peptides 285-300 hFSHR, 216-235 hFSHR, 184-195 hFSHR, 79-89 hFSHR and 15-31 hFSHR were observed to inhibit FSH-induced cAMP production. In summary, this study suggests that discrete, functional domains of the ECD have a role in hormone binding and signal transduction. Region 285-300 has been identified as a novel region crucial for both FSH binding and cAMP generation. PMID- 15350192 TI - Cloning of somatolactin alpha and beta cDNAs in zebrafish and phylogenetic analysis of two distinct somatolactin subtypes in fish. AB - Somatolactin (SL) is a pituitary hormone belonging to the growth hormone/prolactin superfamily, with recognizable homologues in all fish taxa examined to date. Although sequences from most fish share reasonably high sequence identity, several more highly divergent SLs have been reported. Goldfish SL and a second SL protein found in rainbow trout (rtSLP) are remarkably different from each other and also dissimilar to other SLs. It has been unclear whether rtSLP is a recent paralogue restricted to rainbow trout, or reflects a more ancient duplication of the SL gene, and whether it is related to the goldfish sequence. Here we report the cloning of two different zebrafish SL cDNAs, which share only 57.5% nucleotide and 47.7% deduced amino acid identities. One copy, designated zebrafish SLalpha (zfSLalpha), displays a typical range of sequence similarity to most other SLs. The other copy, zebrafish SLbeta (zfSLbeta), shows low identity to most other SLs; surprisingly, it is most similar to the divergent SL sequence from goldfish. The mRNAs of zfSLalpha and zfSLbeta were expressed specifically in two distinct regions of the pars intermedia in zebrafish. Cells expressing zfSLalpha are located at the posterior pars intermedia, bordering the neurohypophysis, whereas zfSLbeta is expressed in the anterior part of the pars intermedia, bordering the pars distalis. Phylogenetic analyses indicate that zfSLbeta, goldfish SL and rtSLP all belong to the SL hormone family; however, along with the genes from eel and catfish, these divergent sequences form a group that is clearly distinct from all other SLs. These results suggest the presence of two distinct SL families, SLalpha and SLbeta, which may trace back to a teleost genome duplication prior to divergence of the cyprinids and salmonids. PMID- 15350193 TI - The Nramp orthologue of Cryptococcus neoformans is a pH-dependent transporter of manganese, iron, cobalt and nickel. AB - Cryptococcus neoformans is an important human opportunistic pathogen and a facultative intracellular parasite, particularly in HIV-infected individuals. Little is known about metal ion transport in this organism. C. neoformans encodes a single member of the Nramp (natural resistance-associated macrophage protein) family of bivalent cation transporters, known as Cramp, which we have cloned and expressed in Xenopus laevis oocytes and Spodoptera frugiperda Sf 21 insect cells. Cramp induces saturable transport of a broad range of bivalent transition series cations, including Mn2+, Fe2+, Co2+ and Ni2+. Maximal cation transport occurs at pH 5.5-6.0, consistent with the proton gradient-based energetics of other Nramp orthologues. Mn2+ transport is diminished in the presence of 140 mM Na+, compatible with a Na+ slippage mechanism proposed for the Saccharomyces cerevisiae Nramp orthologue Smf1p. Cramp resembles Smf1p with respect to predicted membrane topology, substrate specificity and pH dependence, but differs in terms of its apparent affinity for Mn2+ and negligible inhibition by Zn2+. Cramp is the first Nramp orthologue from a fungal pathogen to be functionally characterized. Insights afforded by these findings will allow the formulation of new hypotheses regarding the role of metal ions in the pathophysiology of cryptococcosis. PMID- 15350194 TI - Tissue specificity and regulation of the N-terminal diversity of reticulon 3. AB - Over the last few years, the widely distributed family of reticulons (RTNs) is receiving renewed interest because of the implication of RTN4/Nogo in neurite regeneration. Four genes were identified in mammals and are referred to as RTN1, 2, 3 and the neurite outgrowth inhibitor RTN4/Nogo. In the present paper, we describe the existence of five new isoforms of RTN3 that differ in their N termini, and analysed their tissue distribution and expression in neurons. We redefined the structure of human and murine rtn3 genes, and identified two supplementary exons that may generate up to seven putative isoforms arising by alternative splicing or differential promoter usage. We confirmed the presence of five of these isoforms at the mRNA and protein levels, and showed their preferential expression in the central nervous system. We analysed rtn3 expression in the cerebellum further, and observed increased levels of several of the RTN3 isoforms during cerebellum development and during in vitro maturation of cerebellar granule cells. This pattern of expression paralleled that shown by RTN4/Nogo isoforms. Specifically, RTN3A1 expression was down-regulated upon cell death of cerebellar granule neurons triggered by potassium deprivation. Altogether, our results demonstrate that the rtn3 gene generates multiple isoforms varying in their N-termini, and that their expression is tightly regulated in neurons. These findings suggest that RTN3 isoforms may contribute, by as yet unknown mechanisms, to neuronal survival and plasticity. PMID- 15350195 TI - Glycogen synthase kinase-3 regulates IGFBP-1 gene transcription through the thymine-rich insulin response element. AB - BACKGROUND: Hepatic expression of several gene products involved in glucose metabolism, including phosphoenolpyruvate carboxykinase (PEPCK), glucose-6 phosphatase (G6Pase) and insulin-like growth factor binding protein-1 (IGFBP-1), is rapidly and completely inhibited by insulin. This inhibition is mediated through the regulation of a DNA element present in each of these gene promoters, that we call the Thymine-rich Insulin Response Element (TIRE). The insulin signalling pathway that results in the inhibition of these gene promoters requires the activation of phosphatidylinositol 3-kinase (PI 3-kinase). However, the molecules that connect PI 3-kinase to these gene promoters are not yet fully defined. Glycogen Synthase Kinase 3 (GSK-3) is inhibited following activation of PI 3-kinase. We have shown previously that inhibitors of GSK-3 reduce the activity of two TIRE-containing gene promoters (PEPCK and G6Pase), whose products are required for gluconeogenesis. RESULTS: In this report we demonstrate that in H4IIE-C3 cells, four distinct classes of GSK-3 inhibitor mimic the effect of insulin on a third TIRE-containing gene, IGFBP-1. We identify the TIRE as the minimum requirement for inhibition by these agents, and demonstrate that the target of GSK-3 is unlikely to be the postulated TIRE-binding protein FOXO-1. Importantly, overexpression of GSK-3 in cells reduces the insulin regulation of TIRE activity as well as endogenous IGFBP-1 expression. CONCLUSIONS: These results implicate GSK-3 as an intermediate in the pathway from the insulin receptor to the TIRE. Indeed, this is the first demonstration of an absolute requirement for GSK-3 inhibition in insulin regulation of gene transcription. These data support the potential use of GSK-3 inhibitors in the treatment of insulin resistant states such as Type 2 diabetes mellitus, but suggest that it will be important to identify all TIRE-containing genes to assess potential side effects of these agents. PMID- 15350196 TI - Development of real-time PCR for detection of Mycoplasma hominis. AB - BACKGROUND: Mycoplasma hominis is associated with pelvic inflammatory disease, bacterial vaginosis, post partum fever, sepsis and infections of the central nervous system often leading to serious conditions. Association with development of female infertility has also been suggested, but different publications present different results. We developed a sensitive and fast diagnostic real-time PCR to test clinical samples from women undergoing laparoscopic examination before fertility treatment. To develop a test for the detection and quantification of M. hominis we selected a housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase (gap), as a target. RESULTS: Real-time PCR was optimized to detect 10 copies of M. hominis PG21 genomic DNA. A fluorescence signal was measured for all 20 other M. hominis isolates, and melting curves analysis showed variations in the melting temperature in agreement with sequence variation in the region of the probes. There was no amplification of other mycoplasmal DNA and human DNA. Eighty-three patient cervical swab samples from infertile women were cultured for M. hominis in the BEa medium. Two of the samples (2.4%) were positive after 48 hours of incubation. The real-time PCR detected the same two samples positive, and the DNA concentrations in the clinical specimens were calculated to 37.000 copies/ml and 88.500 copies/ml, respectively. CONCLUSION: The results demonstrate that real time PCR may prove to be a rapid alternative to the traditional cultivation method. Information on bacterial load in genital swabs can be obtained. The assay allowed detection of M. hominis in a closed system reducing the risk of contamination by amplicon carry-over. PMID- 15350197 TI - Determination of the differentially expressed genes in microarray experiments using local FDR. AB - BACKGROUND: Thousands of genes in a genomewide data set are tested against some null hypothesis, for detecting differentially expressed genes in microarray experiments. The expected proportion of false positive genes in a set of genes, called the False Discovery Rate (FDR), has been proposed to measure the statistical significance of this set. Various procedures exist for controlling the FDR. However the threshold (generally 5%) is arbitrary and a specific measure associated with each gene would be worthwhile. RESULTS: Using process intensity estimation methods, we define and give estimates of the local FDR, which may be considered as the probability for a gene to be a false positive. After a global assessment rule controlling the false positive error, the local FDR is a valuable guideline for deciding wether a gene is differentially expressed. The interest of the method is illustrated on three well known data sets. A R routine for computing local FDR estimates from p-values is available at http://www.inapg.fr/ens_rech/mathinfo/recherche/mathematique/outil.html. CONCLUSIONS: The local FDR associated with each gene measures the probability that it is a false positive. It gives the opportunity to compute the FDR of any given group of clones (of the same gene) or genes pertaining to the same regulation network or the same chromosomic region. PMID- 15350198 TI - Handling multiple testing while interpreting microarrays with the Gene Ontology Database. AB - BACKGROUND: The development of software tools that analyze microarray data in the context of genetic knowledgebases is being pursued by multiple research groups using different methods. A common problem for many of these tools is how to correct for multiple statistical testing since simple corrections are overly conservative and more sophisticated corrections are currently impractical. A careful study of the nature of the distribution one would expect by chance, such as by a simulation study, may be able to guide the development of an appropriate correction that is not overly time consuming computationally. RESULTS: We present the results from a preliminary study of the distribution one would expect for analyzing sets of genes extracted from Drosophila, S. cerevisiae, Wormbase, and Gramene databases using the Gene Ontology Database. CONCLUSIONS: We found that the estimated distribution is not regular and is not predictable outside of a particular set of genes. Permutation-based simulations may be necessary to determine the confidence in results of such analyses. PMID- 15350199 TI - GASP: Gapped Ancestral Sequence Prediction for proteins. AB - BACKGROUND: The prediction of ancestral protein sequences from multiple sequence alignments is useful for many bioinformatics analyses. Predicting ancestral sequences is not a simple procedure and relies on accurate alignments and phylogenies. Several algorithms exist based on Maximum Parsimony or Maximum Likelihood methods but many current implementations are unable to process residues with gaps, which may represent insertion/deletion (indel) events or sequence fragments. RESULTS: Here we present a new algorithm, GASP (Gapped Ancestral Sequence Prediction), for predicting ancestral sequences from phylogenetic trees and the corresponding multiple sequence alignments. Alignments may be of any size and contain gaps. GASP first assigns the positions of gaps in the phylogeny before using a likelihood-based approach centred on amino acid substitution matrices to assign ancestral amino acids. Important outgroup information is used by first working down from the tips of the tree to the root, using descendant data only to assign probabilities, and then working back up from the root to the tips using descendant and outgroup data to make predictions. GASP was tested on a number of simulated datasets based on real phylogenies. Prediction accuracy for ungapped data was similar to three alternative algorithms tested, with GASP performing better in some cases and worse in others. Adding simple insertions and deletions to the simulated data did not have a detrimental effect on GASP accuracy. CONCLUSIONS: GASP (Gapped Ancestral Sequence Prediction) will predict ancestral sequences from multiple protein alignments of any size. Although not as accurate in all cases as some of the more sophisticated maximum likelihood approaches, it can process a wide range of input phylogenies and will predict ancestral sequences for gapped and ungapped residues alike. PMID- 15350200 TI - What do evidence-based secondary journals tell us about the publication of clinically important articles in primary healthcare journals? AB - BACKGROUND: We conducted this analysis to determine i) which journals publish high-quality, clinically relevant studies in internal medicine, general/family practice, general practice nursing, and mental health; and ii) the proportion of clinically relevant articles in each journal. METHODS: We performed an analytic survey of a hand search of 170 general medicine, general healthcare, and specialty journals for 2000. Research staff assessed individual articles by using explicit criteria for scientific merit for healthcare application. Practitioners assessed the clinical importance of these articles. Outcome measures were the number of high-quality, clinically relevant studies published in the 170 journal titles and how many of these were published in each of four discipline-specific, secondary "evidence-based" journals (ACP Journal Club for internal medicine and its subspecialties; Evidence-Based Medicine for general/family practice; Evidence Based Nursing for general practice nursing; and Evidence-Based Mental Health for all aspects of mental health). Original studies and review articles were classified for purpose: therapy and prevention, screening and diagnosis, prognosis, etiology and harm, economics and cost, clinical prediction guides, and qualitative studies. RESULTS: We evaluated 60,352 articles from 170 journal titles. The pass criteria of high-quality methods and clinically relevant material were met by 3059 original articles and 1073 review articles. For ACP Journal Club (internal medicine), four titles supplied 56.5% of the articles and 27 titles supplied the other 43.5%. For Evidence-Based Medicine (general/family practice), five titles supplied 50.7% of the articles and 40 titles supplied the remaining 49.3%. For Evidence-Based Nursing (general practice nursing), seven titles supplied 51.0% of the articles and 34 additional titles supplied 49.0%. For Evidence-Based Mental Health (mental health), nine titles supplied 53.2% of the articles and 34 additional titles supplied 46.8%. For the disciplines of internal medicine, general/family practice, and mental health (but not general practice nursing), the number of clinically important articles was correlated withScience Citation Index (SCI) Impact Factors. CONCLUSIONS: Although many clinical journals publish high-quality, clinically relevant and important original studies and systematic reviews, the articles for each discipline studied were concentrated in a small subset of journals. This subset varied according to healthcare discipline; however, many of the important articles for all disciplines in this study were published in broad-based healthcare journals rather than subspecialty or discipline-specific journals. PMID- 15350201 TI - Single group study to evaluate the feasibility and complications of radiofrequency ablation and usefulness of post treatment position emission tomography in lung tumours. AB - BACKGROUND: There is genuine need to develop interventional treatment options for management of lung tumors. Radiofrequency ablation (RFA) is one such alternative being promoted to treat lung tumors recently. Larger studies should help define RFA's further development. Furthermore fluorodeoxyglucose positron emission tomography (PET) has been reported to be an accurate indicator of treatment response in variety of tumors. This study focuses on the evaluating the feasibility of RFA and usefulness of PET scan in lung tumors after RFA procedure. PATIENTS AND METHODS: Between November 1999 and May 2002, 50 patients with primary or metastasis pulmonary tumors underwent RFA procedure. The electrode was guided to the target areas using computerized tomography (CT). Tumors smaller than 3.5 cm were given single RFA, while tumors larger than 3.5 cm received RFA to multiple sites. Maximum 4 lesions or 6 target areas were treated during one operating procedure. Whole body and/or lung PET images were acquired; identical site CT images and chest X-ray were taken 1 week before and after RFA. RESULTS: Of the 50 patients, 17 had single lesions while rest had multiple lesions. Tumors smaller than 3.5 cm were completely dissipated after RFA. In tumors larger than 3.5 cm, the part within 3.5 cm diameter dissipated. While CT showed that tumor image became larger 1 to 2 weeks after RFA procedure. PET demonstrated tumor destruction in 70% cases, compared to 38% in CT. CONCLUSION: The present study shows RFA to be safe and effective treatment option for lung tumors. PET is superior to CT in evaluation the effectiveness of RFA treatment shortly after the procedure. PMID- 15350202 TI - Impairments, activity limitations and participation restrictions: prevalence and associations among persons living with HIV/AIDS in British Columbia. AB - BACKGROUND: To measure the prevalence of and associations among impairments, activity limitations and participation restrictions in persons living with HIV in British Columbia to inform support and care programs, policy and research. METHODS: A cross-sectional population-based sample of persons living with HIV in British Columbia was obtained through an anonymous survey sent to members of the British Columbia Persons With AIDS Society. The survey addressed the experience of physical and mental impairments, and the experience and level of activity limitations and participation restrictions. Associations were measured in three ways: 1) impact of types of impairment on social restriction; 2) impact of specific limitations on social restriction; and 3) independent association of overall impairments and limitations on restriction levels. Logistic regression was used to measure associations with social restriction, while ordinal logistic regression was used to measure associations with a three-category measure of restriction level. RESULTS: The survey was returned by 762 (50.5%) of the BCPWA participants. Over ninety percent of the population experienced one or more impairments, with one-third reporting over ten. Prevalence of activity limitations and participation restrictions was 80.4% and 93.2%, respectively. The presence of social restrictions was most closely associated with mental function impairments (OR: 7.0 for impairment vs. no impairment; 95% CI: 4.7 - 10.4). All limitations were associated with social restriction. Among those with 5 vs. no limitation, 95%CI: 4.9-125.0). Among those with > 200 CD4 cells/mm3, the odds of higher restriction were increased with higher limitation (OR: 2.7 for limitation score of 1-5 vs. no limitation, 95%CI: 1.4 5.1; OR: 8.6 for limitation score > 5 vs. no limitation, 95%CI: 3.9-18.8), as well as by additional number of impairments (OR:1.2 for every additional impairment; 95% CI:1.1-1.3). CONCLUSIONS: This population-based sample of people living with HIV has been experiencing extremely high rates of impairments, activity limitations and participation restrictions. Furthermore, the complex inter-relationships identified amongst the levels reveal lessons for programming, policy and research in terms of the factors that contribute most to a higher quality of life. PMID- 15350203 TI - Protein-polymer nano-machines. Towards synthetic control of biological processes. AB - The exploitation of nature's machinery at length scales below the dimensions of a cell is an exciting challenge for biologists, chemists and physicists, while advances in our understanding of these biological motifs are now providing an opportunity to develop real single molecule devices for technological applications. Single molecule studies are already well advanced and biological molecular motors are being used to guide the design of nano-scale machines. However, controlling the specific functions of these devices in biological systems under changing conditions is difficult. In this review we describe the principles underlying the development of a molecular motor with numerous potential applications in nanotechnology and the use of specific synthetic polymers as prototypic molecular switches for control of the motor function. The molecular motor is a derivative of a TypeI Restriction-Modification (R-M) enzyme and the synthetic polymer is drawn from the class of materials that exhibit a temperature-dependent phase transition.The potential exploitation of single molecules as functional devices has been heralded as the dawn of new era in biotechnology and medicine. It is not surprising, therefore, that the efforts of numerous multidisciplinary teams 12. have been focused in attempts to develop these systems. as machines capable of functioning at the low sub-micron and nanometre length-scales 3. However, one of the obstacles for the practical application of single molecule devices is the lack of functional control methods in biological media, under changing conditions. In this review we describe the conceptual basis for a molecular motor (a derivative of a TypeI Restriction Modification enzyme) with numerous potential applications in nanotechnology and the use of specific synthetic polymers as prototypic molecular switches for controlling the motor function 4. PMID- 15350204 TI - Open Access gains attention in scholar communication. PMID- 15350205 TI - Tuberculosis presenting as immune thrombocytopenic purpura. AB - BACKGROUND: Although various hematologic abnormalities are seen in tuberculosis, immune thrombocytopenic purpura is a rare event. CASE PRESENTATION: We report a case of a 29 year-old male who was presented with immune thrombocytopenia-induced hemoptysis, macroscopic hematuria and generalized petechiae. The patient was found to have clinical, microbiological and radiological evidence of active pulmonary tuberculosis. The immune thrombocytopenic purpura was successfully treated with anti-tuberculous drugs combined with corticosteroids and high dose immune globulin therapy. CONCLUSION: Immune thrombocytopenic purpura can be one of the hematological manifestations of tuberculosis which has a global prevalence with increasing incidence secondary to HIV infection. PMID- 15350206 TI - A weather-driven model of malaria transmission. AB - BACKGROUND: Climate is a major driving force behind malaria transmission and climate data are often used to account for the spatial, seasonal and interannual variation in malaria transmission. METHODS: This paper describes a mathematical biological model of the parasite dynamics, comprising both the weather-dependent within-vector stages and the weather-independent within-host stages. RESULTS: Numerical evaluations of the model in both time and space show that it qualitatively reconstructs the prevalence of infection. CONCLUSION: A process based modelling structure has been developed that may be suitable for the simulation of malaria forecasts based on seasonal weather forecasts. PMID- 15350207 TI - Antibodies from malaria-exposed pregnant women recognize trypsin resistant epitopes on the surface of Plasmodium falciparum-infected erythrocytes selected for adhesion to chondroitin sulphate A. AB - BACKGROUND: The ability of Plasmodium falciparum-infected erythrocytes to adhere to the microvasculature endothelium is thought to play a causal role in malaria pathogenesis. Cytoadhesion to endothelial receptors is generally found to be highly sensitive to trypsinization of the infected erythrocyte surface. However, several studies have found that parasite adhesion to placental receptors can be markedly less sensitive to trypsin. This study investigates whether chondroitin sulphate A (CSA) binding parasites express trypsin-resistant variant surface antigens (VSA) that bind female-specific antibodies induced as a result of pregnancy associated malaria (PAM). METHODS: Fluorescence activated cell sorting (FACS) was used to measure the levels of adult Scottish and Ghanaian male, and Ghanaian pregnant female plasma immunoglobulin G (IgG) that bind to the surface of infected erythrocytes. P. falciparum clone FCR3 cultures were used to assay surface IgG binding before and after selection of the parasite for adhesion to CSA. The effect of proteolytic digestion of parasite erythrocyte surface antigens on surface IgG binding and adhesion to CSA and hyaluronic acid (HA) was also studied. RESULTS: P. falciparum infected erythrocytes selected for adhesion to CSA were found to express trypsin-resistant VSA that are the target of naturally acquired antibodies from pregnant women living in a malaria endemic region of Ghana. However in vitro adhesion to CSA and HA was relatively trypsin sensitive. An improved labelling technique for the detection of VSA expressed by CSA binding isolates has also been described. CONCLUSION: The VSA expressed by CSA binding P. falciparum isolates are currently considered potential targets for a vaccine against PAM. This study identifies discordance between the trypsin sensitivity of CSA binding and surface recognition of CSA selected parasites by serum IgG from malaria exposed pregnant women. Thus, the complete molecular definition of an antigenic P. falciparum erythrocyte surface protein that can be used as a malaria in pregnancy vaccine has not yet been achieved. PMID- 15350208 TI - Microarrays in ecological research: a case study of a cDNA microarray for plant herbivore interactions. AB - BACKGROUND: Microarray technology allows researchers to simultaneously monitor changes in the expression ratios (ERs) of hundreds of genes and has thereby revolutionized most of biology. Although this technique has the potential of elucidating early stages in an organism's phenotypic response to complex ecological interactions, to date, it has not been fully incorporated into ecological research. This is partially due to a lack of simple procedures of handling and analyzing the expression ratio (ER) data produced from microarrays. RESULTS: We describe an analysis of the sources of variation in ERs from 73 hybridized cDNA microarrays, each with 234 herbivory-elicited genes from the model ecological expression system, Nicotiana attenuata, using procedures that are commonly used in ecologic research. Each gene is represented by two independently labeled PCR products and each product was arrayed in quadruplicate. We present a robust method of normalizing and analyzing ERs based on arbitrary thresholds and statistical criteria, and characterize a "norm of reaction" of ERs for 6 genes (4 of known function, 2 of unknown) with different ERs as determined across all analyzed arrays to provide a biologically-informed alternative to the use of arbitrary expression ratios in determining significance of expression. These gene-specific ERs and their variance (gene CV) were used to calculate array based variances (array CV), which, in turn, were used to study the effects of array age, probe cDNA quantity and quality, and quality of spotted PCR products as estimates of technical variation. Cluster analysis and a Principal Component Analysis (PCA) were used to reveal associations among the transcriptional "imprints" of arrays hybridized with cDNA probes derived from mRNA from N. attenuata plants variously elicited and attacked by different herbivore species and from three congeners: N. quadrivalis, N. longiflora and N. clevelandii. Additionally, the PCA revealed the contribution of individual gene ERs to the associations among arrays. CONCLUSIONS: While the costs of 'boutique' array fabrication are rapidly declining, familiar methods for the analysis of the data they create are still missing. The case history illustrated here demonstrates the ease with which this powerful technology can be adapted to ecological research. PMID- 15350209 TI - ISSR and AFLP analysis of the temporal and spatial population structure of the post-fire annual, Nicotiana attenuata, in SW Utah. AB - BACKGROUND: The native annual tobacco, Nicotiana attenuata, is found primarily in large ephemeral populations (typically for less than 3 growing seasons) after fires in sagebrush and pinyon-juniper ecosystems and in small persistent populations (for many growing seasons) in isolated washes typically along roadsides throughout the Great Basin Desert of the SW USA. This distribution pattern is due to its unusual germination behavior. Ephemeral populations are produced by the germination of dormant seeds from long-lived seed banks which are stimulated to germinate by a combination of unidentified positive cues found in wood smoke and the removal of inhibitors leached from the unburned litter of the dominant vegetation. Persistent populations may result where these inhibitors do not exist, as in washes or along disturbed roadsides. To determine if this germination behavior has influenced population structure, we conducted an AFLP (244 individuals), ISSR (175 individuals) and ISSR+ AFLP (175 individuals) analysis on plants originating from seed collected from populations growing in 11 wash and burns over 11 years from the SW USA. RESULTS: Genetic variance as measured by both ISSR and AFLP markers was low among sites and comparatively higher within populations. Cluster analysis of the Utah samples with samples collected from Arizona, California, and Oregon as out-groups also did not reveal patterns. AMOVA analysis of the combined AFLP and ISSR data sets yielded significantly low genetic differentiation among sites (Phict), moderate among populations within sites (Phisc) and higher genetic differentiation within populations (Phist). CONCLUSIONS: We conclude that the seed dormancy of this post fire annual and its resulting age structure in conjunction with natural selection processes are responsible for significantly low among sites and comparatively high within-population genetic variation observed in this species. PMID- 15350210 TI - The relationship between educational level and bone mineral density in postmenopausal women. AB - BACKGROUND: This study describes the influence of educational level on bone mineral density (BMD) and investigating the relationship between educational level and bone mineral density in postmenopausal women. METHODS: A total of 569 postmenopausal women, from 45 to 86 years of age (mean age of 60.43 +/- 7.19 years) were included in this study. A standardized interview was used at the follow-up visit to obtain information on demographic, life-style, reproductive and menstrual histories such as age at menarche, age at menopause, number of pregnancies, number of abortions, duration of menopause, duration of fertility, and duration of lactation. Patients were separated into four groups according to the level of education, namely no education (Group 1 with 209 patients), elementary (Group 2 with 222 patients), high school (Group 3 with 79 patients), and university (Group 4 with 59 patients). RESULTS: The mean ages of groups were 59.75 +/- 7.29, 61.42 +/- 7.50, 60.23 +/- 7.49, and 58.72 +/- 7.46, respectively. Spine BMD was significant lower in Group 1 than that of other groups (p < 0.05). Trochanter and ward's triangle BMD were the highest in Group 4 and there was a significant difference between Group 1 and 4 (p < 0.05). The prevalence of osteoporosis showed an inverse relationship with level of education, ranging from 18.6% for the most educated to 34.4% for the no educated women (p < 0.05). Additionally, there was a significant correlation between educational level and spine BMD (r = 0.20, p < 0.01), trochanter BMD (r = 0.13, p < 0.01), and ward's BMD (r = 0.14, p < 0.01). CONCLUSIONS: The results of the study suggest that there is a significant correlation between educational level and BMD. Losses in BMD for women of lower educational level tend to be relatively high, and losses in spine and femur BMD showed a decrease with increasing educational level. PMID- 15350211 TI - Coactivators p300 and PCAF physically and functionally interact with the foamy viral trans-activator. AB - BACKGROUND: Foamy virus Bel1/Tas trans-activators act as key regulators of gene expression and directly bind to Bel1 response elements (BRE) in both the internal and the 5'LTR promoters leading to strong transcriptional trans-activation. Cellular coactivators interacting with Bel1/Tas are unknown to date. RESULTS: Transient expression assays, co-immunoprecipitation experiments, pull-down assays, and Western blot analysis were used to demonstrate that the coactivator p300 and histone acetyltransferase PCAF specifically interact with the retroviral trans-activator Bel1/Tas in vivo. Here we show that the Bel1/Tas-mediated trans activation was enhanced by the coactivator p300, histone acetyltransferases PCAF and SRC-1 based on the crucial internal promoter BRE. The Bel1/Tas-interacting region was mapped to the C/H1 domain of p300 by co-immunoprecipitation and pull down assays. In contrast, coactivator SRC-1 previously reported to bind to the C terminal domain of p300 did not directly interact with the Bel1 protein but nevertheless enhanced Bel1/Tas-mediated trans-activation. Cotransfection of Bel1/Tas and p300C with an expression plasmid containing the C/H1domain partially inhibited the p300C-driven trans-activation. CONCLUSIONS: Our data identify p300 and PCAF as functional partner molecules that directly interact with Bel1/Tas. Since the acetylation activities of the three coactivators reside in or bind to the C-terminal regions of p300, a C/H1 expression plasmid was used as inhibitor. This is the first report of a C/H1 domain-interacting retroviral trans-activator capable of partially blocking the strong Bel1/Tas-mediated activation of the C terminal region of coactivator p300. The potential mechanisms and functional roles of the three histone and factor acetyltransferases p300, PCAF, and SRC-1 in Bel1/Tas-mediated trans-activation are discussed. PMID- 15350212 TI - Regulation of protein kinases; controlling activity through activation segment conformation. AB - There are currently at least forty-six unique protein kinase crystal structures, twenty-four of which are available in an active state. Here we examine these structures using a structural bioinformatics approach to understand how the conformation of the activation segment controls kinase activity. PMID- 15350213 TI - Salmonella typhimurium recognizes a chemically distinct form of the bacterial quorum-sensing signal AI-2. AB - Bacterial populations use cell-cell communication to coordinate community-wide regulation of processes such as biofilm formation, virulence, and bioluminescence. This phenomenon, termed quorum sensing, is mediated by small molecule signals known as autoinducers. While most autoinducers are species specific, autoinducer-2 (AI-2), first identified in the marine bacterium Vibrio harveyi, is produced and detected by many Gram-negative and Gram-positive bacteria. The crystal structure of the V. harveyi AI-2 signaling molecule bound to its receptor protein revealed an unusual furanosyl borate diester. Here, we present the crystal structure of a second AI-2 signal binding protein, LsrB from Salmonella typhimurium. We find that LsrB binds a chemically distinct form of the AI-2 signal, (2R,4S)-2-methyl-2,3,3,4-tetrahydroxytetrahydrofuran (R-THMF), that lacks boron. Our results demonstrate that two different species of bacteria recognize two different forms of the autoinducer signal, both derived from 4,5 dihydroxy-2,3-pentanedione (DPD), and reveal new sophistication in the chemical lexicon used by bacteria in interspecies signaling. PMID- 15350214 TI - Structure of a human inositol 1,4,5-trisphosphate 3-kinase: substrate binding reveals why it is not a phosphoinositide 3-kinase. AB - Mammalian cells produce a variety of inositol phosphates (InsPs), including Ins(1,4,5)P3 that serves both as a second messenger and as a substrate for inositol polyphosphate kinases (IPKs), which further phosphorylate it. We report the structure of an IPK, the human Ins(1,4,5)P3 3-kinase-A, both free and in complexes with substrates and products. This enzyme catalyzes transfer of a phosphate from ATP to the 3-OH of Ins(1,4,5)P3, and its X-ray crystal structure provides a template for understanding a broad family of InsP kinases. The catalytic domain consists of three lobes. The N and C lobes bind ATP and resemble protein and lipid kinases, despite insignificant sequence similarity. The third lobe binds inositol phosphate and is a unique four-helix insertion in the C lobe. This lobe embraces all of the phosphates of Ins(1,4,5)P3 in a positively charged pocket, explaining the enzyme's substrate specificity and its inability to phosphorylate PtdIns(4,5)P2, the membrane-resident analog of Ins(1,4,5)P3. PMID- 15350215 TI - Crystal structure of the catalytic core of inositol 1,4,5-trisphosphate 3-kinase. AB - Soluble inositol polyphosphates are ubiquitous second messengers in eukaryotes, and their levels are regulated by an array of specialized kinases. The structure of an archetypal member of this class, inositol 1,4,5-trisphosphate 3-kinase (IP3K), has been determined at 2.2 angstroms resolution in complex with magnesium and adenosine diphosphate. IP3K contains a catalytic domain that is a variant of the protein kinase superfamily, and a novel four-helix substrate binding domain. The two domains are in an open conformation with respect to each other, suggesting that substrate recognition and catalysis by IP3K involves a dynamic conformational cycle. The unique helical domain of IP3K blocks access to the active site by membrane-bound phosphoinositides, explaining the structural basis for soluble inositol polyphosphate specificity. PMID- 15350216 TI - Distinct roles for JNK1 and JNK2 in regulating JNK activity and c-Jun-dependent cell proliferation. AB - Different c-Jun N-terminal kinases (JNKs) are activated by a plethora of signals and phosphorylate substrates such as c-Jun, which is required for efficient cell cycle progression. Although JNK1 and JNK2 were shown to differentially regulate fibroblast proliferation, the underlying mechanistic basis remains unclear. We found that Jnk2-/- fibroblasts exit G1 and enter S phase earlier than wild-type counterparts, while Jnk1-/- cells show the inverse phenotype. Moreover, Jnk2-/- erythroblasts also exhibit a proliferative advantage. JNK2 deficiency results in elevated c-Jun phosphorylation and stability, whereas the absence of JNK1 reduces c-Jun phosphorylation and stability. Re-expression of JNK2 in Jnk2-/- cells reverses the JNK2 null phenotype, whereas ectopic expression of JNK1 augments it. JNK2 is preferentially bound to c-Jun in unstimulated cells, thereby contributing to c-Jun degradation. In contrast, JNK1 becomes the major c-Jun interacting kinase after cell stimulation. These data provide mechanistic insights into the distinct roles of different JNK isoforms. PMID- 15350217 TI - Crystal structure of IIGP1: a paradigm for interferon-inducible p47 resistance GTPases. AB - Interferon-inducible p47 GTPases are critical mediators of cell-autonomous resistance against several intracellular pathogens. Here we present the first crystal structure of a member of this novel GTPase family, IIGP1, in its nucleotide-free, GDP-, and GppNHp-bound form. The structure shows a Ras-like G domain between an N-terminal three-helix bundle and a complex system of C terminal helices and loops. Sequence comparison and secondary structure prediction suggest the IIGP1 structure to be a valid model for the p47 GTPase family. The IIGP1 crystals contain a noncrystallographic dimer. We show that the dimer is required for cooperative GTP hydrolysis and GTP-dependent oligomerization of IIGP1. We also present the GDP- and GppNHp-bound monomeric structures of two dimer interface mutants. Our structures direct approaches to the analysis of the catalytic mechanism of IIGP1 and provide a coherent basis for structure-function studies aimed at elucidating the mechanistic basis of pathogen resistance caused by these enigmatic GTPases. PMID- 15350218 TI - WNK1 phosphorylates synaptotagmin 2 and modulates its membrane binding. AB - WNK (with no lysine [K]) protein kinases were named for their unique active site organization. Mutations in WNK1 and WNK4 cause a familial form of hypertension by undefined mechanisms. Here, we report that WNK1 selectively binds to and phosphorylates synaptotagmin 2 (Syt2) within its calcium binding C2 domains. Endogenous WNK1 and Syt2 coimmunoprecipitate and colocalize on a subset of secretory granules in INS-1 cells. Phosphorylation by WNK1 increases the amount of Ca2+ required for Syt2 binding to phospholipid vesicles; mutation of threonine 202, a WNK1 phosphorylation site, partially prevents this change. These findings suggest that phosphorylation of Syts by WNK1 can regulate Ca2+ sensing and the subsequent Ca2+-dependent interactions mediated by Syt C2 domains. These findings provide a biochemical mechanism that could lead to the retention or insertion of proteins in the plasma membrane. Interruption of this regulatory pathway may disturb membrane events that regulate ion balance. PMID- 15350219 TI - GM1-ganglioside-mediated activation of the unfolded protein response causes neuronal death in a neurodegenerative gangliosidosis. AB - GM1-ganglioside (GM1) is a major sialoglycolipid of neuronal membranes that, among other functions, modulates calcium homeostasis. Excessive accumulation of GM1 due to deficiency of lysosomal beta-galactosidase (beta-gal) characterizes the neurodegenerative disease GM1-gangliosidosis, but whether the accumulation of GM1 is directly responsible for CNS pathogenesis was unknown. Here we demonstrate that activation of an unfolded protein response (UPR) associated with the upregulation of BiP and CHOP and the activation of JNK2 and caspase-12 leads to neuronal apoptosis in the mouse model of GM1-gangliosidosis. GM1 loading of wild type neurospheres recapitulated the phenotype of beta-gal-/- cells and activated this pathway by depleting ER calcium stores, which ultimately culminated in apoptosis. Activation of UPR pathways did not occur in mice double deficient for beta-gal and ganglioside synthase, beta-gal-/-/GalNAcT-/-, which do not accumulate GM1. These findings suggest that the UPR can be induced by accumulation of the sialoglycolipid GM1 and this causes a novel mechanism of neuronal apoptosis. PMID- 15350220 TI - Degradation of misfolded proteins prevents ER-derived oxidative stress and cell death. AB - A variety of debilitating diseases including diabetes, Alzheimer's, Huntington's, Parkinson's, and prion-based diseases are linked to stress within the endoplasmic reticulum (ER). Using S. cerevisiae, we sought to determine the relationship between protein misfolding, ER stress, and cell death. In the absence of ERV29, a stress-induced gene required for ER associated degradation (ERAD), misfolded proteins accumulate in the ER leading to persistent ER stress and subsequent cell death. Cells alleviate ER stress through the unfolded protein response (UPR); however, if stress is sustained the UPR contributes to cell death by causing the accumulation of reactive oxygen species (ROS). ROS are generated from two sources: the UPR-regulated oxidative folding machinery in the ER and mitochondria. Our results demonstrate a direct mechanism(s) by which misfolded proteins lead to cellular damage and death. PMID- 15350221 TI - RNA displacement and resolution of the transcription bubble during transcription by T7 RNA polymerase. AB - Unlike DNA polymerases, RNA polymerases (RNAPs) must displace the nascent product from the template and restore the DNA to duplex form after passage of the transcription complex. To accomplish this, RNAPs establish a locally denatured "bubble" that encloses a short RNA:DNA hybrid. As the polymerase advances along the template, the RNA is displaced at the trailing edge of the bubble and the two DNA strands are reannealed. Structural analyses have revealed a number of elements that are likely to be involved in this process in T7 RNAP. In this work, we used genetic and biochemical methods to explore the roles of these elements during the transition from an initiation complex to an elongation complex. The results indicate that the transition is a multistep process and reveal a critical role for the nontemplate strand of the DNA. PMID- 15350222 TI - A RecA filament capping mechanism for RecX protein. AB - The RecX protein is a potent inhibitor of RecA protein activities. RecX functions by specifically blocking the extension of RecA filaments. In vitro, this leads to a net disassembly of RecA protein from circular single-stranded DNA. Based on multiple observations, we propose that RecX has a RecA filament capping activity. This activity has predictable effects on the formation and disassembly of RecA filaments. In vivo, the RecX protein may limit the length of RecA filaments formed during recombinational DNA repair and other activities. RecX protein interacts directly with RecA protein, but appears to interact in a functionally significant manner only with RecA filaments bound to DNA. PMID- 15350223 TI - Polo-like kinase-1 controls recovery from a G2 DNA damage-induced arrest in mammalian cells. AB - DNA damage triggers multiple checkpoint pathways to arrest cell cycle progression. Less is known about the mechanisms that allow resumption of the cell cycle once checkpoint signaling is silenced. Here we show that while in undamaged cells several redundant pathways can promote the onset of mitosis, this redundancy is lost in cells recovering from a DNA damage-induced arrest. We demonstrate that Plk1 is crucial for mitotic entry following recovery from DNA damage. However, Plk1 is no longer required in cells depleted of Wee1, and we could show that Plk1 is involved in the degradation of Wee1 at the onset of mitosis. Thus, our data show that the cell cycle machinery is reset in response to DNA damage and that cells become critically dependent on Plk1-mediated degradation of Wee1 for their recovery. PMID- 15350224 TI - Structural basis of heteromeric smad protein assembly in TGF-beta signaling. AB - The formation of protein complexes between phosphorylated R-Smads and Smad4 is a central event in the TGF-beta signaling pathway. We have determined the crystal structure of two R-Smad/Smad4 complexes, Smad3/Smad4 to 2.5 angstroms, and Smad2/Smad4 to 2.7 angstroms. Both complexes are heterotrimers, comprising two phosphorylated R-Smad subunits and one Smad4 subunit, a finding that was corroborated by isothermal titration calorimetry and mutational studies. Preferential formation of the R-Smad/Smad4 heterotrimer over the R-Smad homotrimer is largely enthalpy driven, contributed by the unique presence of strong electrostatic interactions within the heterotrimeric interfaces. The study supports a common mechanism of Smad protein assembly in TGF-beta superfamily signaling. PMID- 15350225 TI - Itch E3 ligase-mediated regulation of TGF-beta signaling by modulating smad2 phosphorylation. AB - Protein ubiquitination has been implicated in the intracellular biochemical events transduced by TGF-beta receptor via different mechanisms including the degradation of Smads or their binding proteins. Here we show that loss of Itch E3 ligase in mouse embryonic fibroblasts (MEFs) results in reduced susceptibility of TGF-beta-induced cell growth arrest and decreased phosphorylation of Smad2, without apparent alteration in protein levels for Smad2, Smad4, and Smad7 in Itch /- MEFs. Itch promotes ubiquitination of Smad2 and augments Smad2 phosphorylation that requires an intact ligase activity of Itch. Moreover, Itch facilitates complex formation between TGF-beta receptor and Smad2 and enhances TGF-beta induced transcription. This study reveals a previously unrecognized positive TGF beta signaling pathway via proteolysis-independent ubiquitination. PMID- 15350226 TI - Three-dimensional structure of the native spliceosome by cryo-electron microscopy. AB - Splicing of pre-mRNA occurs in a multicomponent macromolecular machine--the spliceosome. The spliceosome can be assembled in vitro by a stepwise assembly of a number of snRNPs and additional proteins on exogenously added pre-mRNA. In contrast, splicing in vivo occurs in preformed particles where endogenous pre mRNAs are packaged with all five spliceosomal U snRNPs (penta-snRNP) together with other splicing factors. Here we present a three-dimensional image reconstruction by cryo-electron microscopy of native spliceosomes, derived from cell nuclei, at a resolution of 20 angstroms. The structure revealed an elongated globular particle made up of two distinct subunits connected to each other leaving a tunnel in between. We show here that the larger subunit is a suitable candidate to accommodate the penta-snRNP, and that the tunnel could accommodate the pre-mRNA component of the spliceosome. The features this structure reveals provide new insight into the global architecture of the native splicing machine. PMID- 15350227 TI - The rise (and fall?) of coronary care nurses. PMID- 15350228 TI - Seeking an explanation for the poor uptake of in-hospital AED programs. AB - The automated external defibrillator (AED) has been adopted by emergency service personnel as a first-line intervention in the management of out-of-hospital cardiac arrest (OHCA). AED leads to more successful Advanced Cardiac Life Support; consequently, national resuscitation organisations worldwide have recommended that nurses and doctors also integrate AEDs as a component of their basic life-support response to cardiac arrest. Despite these recommendations, the implementation of AED programs within hospitals has been generally sporadic and isolated. A continuation of this situation will most likely disturb and perplex nurses and patients, as they are key stakeholders with respect to upholding recommended BLS practices. In the absence of any explanation from change agents within hospitals, this paper seeks, by way of a pilot study and a review of the literature, to identify the extent of the problem and identify factors contributing to the relatively slow uptake of this device. We argue that nurses and other first responders to in-hospital cardiac arrest (CA) have much to gain, in the context of Occupational Health Safety and Welfare (OHS and W), from ready access to AEDs. Cost factors are also considered, with initial cost of AED purchase likely to be a major concern for managers of hospital budgets. The issues we discuss in this paper clearly support the need for further research to (a) explain the nature of public hospital resistance to AEDs and (b) to consider whether AEDs will provide practical advantages to public hospitals from an occupational, social and economic perspective. PMID- 15350229 TI - Partners of ICD patients--an exploratory study of their experiences. AB - BACKGROUND: The experiences of partners who care for and support the needs of a loved one with an implantable cardiac defibrillator (ICD) remain largely unknown within Europe. AIMS: This study explored the nature of partner's experiences from the pre-ICD implantation phase up until a maximum of 20 months at home. METHODS: Eight partners cohabiting with a recipient of an ICD were interviewed using a semistructured schedule. All interviews were transcribed and content analysed for emerging categories and themes. FINDINGS: Four themes representing 11 categories conceptualised the partners' experiences, these included: Acknowledging the patient's need for the device, reactions to the device, safeguarding the patient, and returning to normality. CONCLUSION: This qualitative study suggests that partners of ICD recipients progress through various difficult and adaptive stages when learning how best to support the patient. A point is reached when most are able to assume control and normalise their lives. This transformation is slow; however, to improve this process and empower partners, nurses should provide relevant information and include them in decisions affecting the patient. Further research into the unique needs of partners, which includes international perspectives, would be significant in developing practice and theory in this area. PMID- 15350230 TI - Peer support. An under-recognized resource in cardiac recovery. AB - BACKGROUND: Cardiovascular disease remains the leading cause of mortality and premature death in western societies. Thus, rates of interventions such as coronary artery bypass surgery are continuing to grow. Health care reform and initiatives to reduce health care expenditures have resulted in early patient discharge from hospital following cardiac surgery. With subsequent cutbacks in nursing support and community-based care, patients are leaving hospital less prepared and supported to deal with the changes that occur during the first weeks of recovery. AIMS: To examine the theoretical assumptions that support the contention that peer support is an under-utilized resource for patients who are recovering from cardiac surgery and the challenges to evaluating peer support interventions. METHODS: A review of current literature, which focuses on cardiac surgery recovery, transitions, social support, and peer support interventions. RESULTS: Peer support (lay assistance from individuals who possess experiential knowledge and similar characteristics), a form of social support, is a viable and potentially sustainable mechanism to put in place during transitional life events such as recovery from cardiac surgery. CONCLUSIONS: Further investigation is needed of peer support interventions for cardiac surgery patients. Specifically, investigations of the influence of peer support interventions on recovery and health outcomes are necessary in this patient population. Yet, challenges exist to undertaking well-designed investigations of social interventions such as peer support. PMID- 15350231 TI - Psychological morbidity after myocardial infarction in an area of deprivation in the UK: evaluation of a self-help package. AB - BACKGROUND: Psychological morbidity after an acute myocardial infarction (AMI) is known to be common, but can be addressed by appropriate rehabilitation. The area in which this research was conducted experiences high rates of deprivation and of coronary heart disease and limited access to hospital-based rehabilitation. Responding to concern about psychological needs of AMI patients, a self-help package was introduced and evaluated alongside standard hospital-based cardiac rehabilitation. AIMS: To evaluate the impact of a home-based self-help package (the Heart Manual), alongside existing cardiac rehabilitation provision, on psychological morbidity and health status after AMI. A secondary aim was to assess the suitability of the Heart Manual for older patients aged over 80 years. METHODS: A controlled observational study, comparing two cohorts of patients discharged from hospital after AMI. The intervention group was given the self help package in addition to standard care. The control group received standard care alone. Outcome measures used were the Hospital Anxiety and Depression Scale and the EuroQol. RESULTS: The intervention group showed significant improvement in anxiety and depression scores after 3 months and nonsignificant improvement in general health status. Patients who attended hospital-based rehabilitation classes, and those aged over 80 years, also benefited from the intervention. CONCLUSION: A home-based self-help rehabilitation package is an effective tool alongside hospital-based rehabilitation classes and can be given to all age groups. PMID- 15350232 TI - International comparison of factors associated with delay in presentation for AMI treatment. AB - BACKGROUND: Prehospital delay in response to acute myocardial infarction (AMI) symptoms is well documented in the US and Europe, but little is known about it in Asian countries where cardiovascular disease is increasing. AIMS: We conducted an observational study of delay times and factors associated with hospital presentation times in 595 patients with AMI from the US, England, Japan and South Korea. METHODS: Patients were interviewed about responses to symptoms within 72 h of hospital admission and the medical records were reviewed. RESULTS: The proportions of patients with delay times of 1 h or less were: US--23%, Korea- 18%, England--15% and Japan--8%. In the US and England when others present at symptom onset called an ambulance patients presented two to three times sooner. Independent predictors of presentation within an hour of symptom onset were attribution of symptoms to the heart and not waiting for symptoms to go away. CONCLUSION: Similar education about the need to seek treatment early in response to AMI symptoms may be applicable in Western and Eastern industrialised populations. PMID- 15350233 TI - The contribution of optimism and quality of life to depression in an acute coronary syndrome population. AB - BACKGROUND: Numerous longitudinal studies have revealed that depression following an acute cardiac event poses a risk factor for poor cardiac outcomes. It is therefore important to identify modifiable predictors of depression in order to develop a variety of interventions with this population. AIMS: The aim of the present research was to determine whether the relationship between optimism and depressive symptoms was mediated by self-reported quality of life (QOL) in acute coronary syndrome patients. METHODS: Two weeks following hospital discharge (Time 1) 59 participants completed a self-report questionnaire. Four weeks later (Time 2), 49 of these participants completed the same questionnaire. RESULTS: At Time 1, the relationship between optimism and depressive symptoms was partially mediated by functional QOL and symptom QOL. Furthermore, the relationship between Time 1 optimism and Time 2 depressive symptoms was partially mediated by Time 1 functional QOL. When each of the Time 1 variables were used to predict Time 2 depressive symptoms, only optimism continued to predict depressive symptoms over and above the influence of Time 1 depressive symptoms and other covariates. CONCLUSION: These findings suggest the underlying importance of optimism in influencing depressive symptoms in acute coronary syndrome patients, and indicate that optimism and perceptions of functional QOL may be a possible rehabilitation target for this population. PMID- 15350234 TI - The phenomena of participation and non-participation in health care--experiences of patients attending a nurse-led clinic for chronic heart failure. AB - BACKGROUND: Patient participation is stressed in the health care acts of many western countries yet a common definition of the concept is lacking. The understanding of experiences of patients with chronic heart failure (CHF) who attend nurse-led specialist clinics, a form of care suggested as beneficiary to this group, may promote a better understanding of participation. AIM: To investigate the meanings of participation and non-participation as experienced by patients living with CHF. METHODS: Narrative interviews analysed in the phenomenological hermeneutic tradition inspired by Ricoeur where the interpretation is made in the hermeneutic circle, explaining and understanding the experienced phenomena. FINDINGS: Participation was experienced as to "be confident", "comprehend" and "seek and maintain a sense of control". Non participation was experienced as to "not understand", "not be in control", "lack a relationship" and "not be accountable". The findings indicate that the experiences of participation and non-participation can change over time and phases of the disease and treatment. CONCLUSION: The study suggests an extended view on the concept of participation. Patients' experiences of participation in health care can vary and should therefore be an issue for dialogue between nurses and patients with CHF in nurse-led specialist clinics. PMID- 15350235 TI - Anxiety is not manifested by elevated heart rate and blood pressure in acutely ill cardiac patients. AB - BACKGROUND: Accurate assessment of anxiety in cardiac patients is important because anxiety is associated with adverse outcomes. Clinicians often use heart rate and blood pressure as indicators of anxiety; however, little is known about whether these measures accurately reflect anxiety in acutely ill patients. AIMS: The purpose of this study was to determine whether heart rate and blood pressure were related to level of anxiety in patients with chronic advanced heart failure (HF), patients with acute myocardial infarction (AMI), and healthy individuals. METHODS AND RESULTS: In this descriptive, correlational study, anxiety, heart rate, and blood pressure were measured at the same time in three groups of individuals: (1) 54 patients hospitalized for AMI; (2) 32 patients with chronic advanced HF; and (3) 31 healthy individuals. State anxiety was measured using the anxiety subscale of the Brief Symptom Inventory. Heart rate and blood pressure data were collected immediately prior to the anxiety assessment. Data were collected in the outpatient setting for patients with HF and healthy individuals. For patients with AMI, data were collected a mean of 48+/-33 h after admission. There were no correlations between anxiety and heart rate or diastolic blood pressure. Higher anxiety was associated with lower systolic blood pressure in patients with AMI (r=-0.23, P<0.05) and in healthy individuals (r=-0.27, P<0.05). CONCLUSION: Elevated heart rate and blood pressure do not accurately reflect level of anxiety as reported by patients with HF or AMI and healthy individuals, and thus cannot be used to assess anxiety in acutely ill patients. Clinicians who use changes in heart rate or blood pressure as indicators of anxiety may fail to recognize and treat anxiety, placing their patients at high risk for both immediate and long-term complications. PMID- 15350236 TI - Cardiovascular nursing in RN and higher education in Swedish universities: a national survey. AB - Cardiovascular nursing (CVN) is rapidly developing and has accumulated a large amount of evidence to support interventions aimed at reducing suffering and hastening recovery. However, knowledge of the extent and content of CVN training in Sweden is lacking. The aim of this study was therefore to identify and describe CVN in the Swedish RN education as well as in higher education, with reference to type of course and credits, content, area and target group. A nationwide survey was carried out in Sweden at all university level nursing schools (N=26) by means of a 25-item questionnaire, which was analysed by descriptive statistics. The findings show that 69% and 23% of the schools awarded 0-1 credits [0-1.5 European Credit Transfer and Accumulation System (ECTS)] and 2 3 credits (3-4.5 ECTS), respectively for cardiology/CVN in the RN education. Target areas outside the hospital setting reported by 23% and 19% of nursing schools were primary health care and community care, respectively. Special target groups in addition to the general public were the elderly (42%) and women (58%). Advanced courses in CVN comprising 10-40 credits (15-60 ECTS) were held by 27% of nursing schools, but no school had a specialist or Master level education. Important educational implications from the study of the RN education are the establishment of a minimum credit figure and to reach out into primary health care. PMID- 15350237 TI - Time is of the essence. AB - Timing is essential to human behaviour, but the neural mechanisms underlying time perception are still unclear. New findings from a brain-imaging study by Coull et al. show that activity in a network of motor-related areas varies parametrically with attention to time. Given that a system in which timing is important (but not the primary function) is recruited when temporal judgements are required, we should perhaps reassess the notion of a dedicated timing system in the brain. PMID- 15350238 TI - Asymmetries in face and brain related to emotion. AB - Research on the neural substrates of emotion has found evidence for cortical asymmetries for aspects of emotion. A recent article by Nicholls et al. has used a new imaging method to interrogate facial movement in 3D to assess possible asymmetrical action during expressions of happiness and sadness. Greater left sided movement, particularly during expressions of sadness was observed. These findings have implications for understanding hemispheric differences in emotion and lend support to the notion that aspects of emotion processing might be differentially localized in the two hemispheres. PMID- 15350239 TI - Characterizing the evolutionary dynamics of language. AB - In a recent article Mitchener and Nowak present a model of the evolutionary dynamics of language. The model exhibits regular and chaotic oscillations in changes to the proportions of grammars spoken in a population over the course of evolution. These oscillations are within the purview of evolutionary game theory, but they suggest the lack of an evolutionarily stable strategy. Implications for self-organization across scales of adaptation are discussed. PMID- 15350240 TI - A unifying view of the basis of social cognition. AB - In this article we provide a unifying neural hypothesis on how individuals understand the actions and emotions of others. Our main claim is that the fundamental mechanism at the basis of the experiential understanding of others' actions is the activation of the mirror neuron system. A similar mechanism, but involving the activation of viscero-motor centers, underlies the experiential understanding of the emotions of others. PMID- 15350241 TI - Psychological essentialism in children. AB - Psychological essentialism is the idea that certain categories, such as "lion" or "female", have an underlying reality that cannot be observed directly. Where does this idea come from? This article reviews recent evidence suggesting that psychological essentialism is an early cognitive bias. Young children look beyond the obvious in many converging ways: when learning words, generalizing knowledge to new category members, reasoning about the insides of things, contemplating the role of nature versus nurture, and constructing causal explanations. These findings argue against the standard view of children as concrete thinkers, instead claiming that children have an early tendency to search for hidden, non obvious features. PMID- 15350242 TI - Action sets and decisions in the medial frontal cortex. AB - Activations in human dorsomedial frontal and cingulate cortices are often present in neuroimaging studies of decision making and action selection. Interpretations have emphasized executive control, movement sequencing, error detection and conflict monitoring. Recently, however, experimental approaches, using lesions, inactivation, and cell recording, have suggested that these are just components of the areas' functions. Here we review these results and integrate them with those from neuroimaging. A medial superior frontal gyrus (SFG) region centred on the pre-supplementary motor area (pre-SMA) is involved in the selection of action sets whereas the anterior cingulate cortex (ACC) has a fundamental role in relating actions to their consequences, both positive reinforcement outcomes and errors, and in guiding decisions about which actions are worth making. PMID- 15350243 TI - Organization, development and function of complex brain networks. AB - Recent research has revealed general principles in the structural and functional organization of complex networks which are shared by various natural, social and technological systems. This review examines these principles as applied to the organization, development and function of complex brain networks. Specifically, we examine the structural properties of large-scale anatomical and functional brain networks and discuss how they might arise in the course of network growth and rewiring. Moreover, we examine the relationship between the structural substrate of neuroanatomy and more dynamic functional and effective connectivity patterns that underlie human cognition. We suggest that network analysis offers new fundamental insights into global and integrative aspects of brain function, including the origin of flexible and coherent cognitive states within the neural architecture. PMID- 15350244 TI - Adaptive neural models of queuing and timing in fluent action. AB - In biological cognition, specialized representations and associated control processes solve the temporal problems inherent in skilled action. Recent data and neural circuit models highlight three distinct levels of temporal structure: sequence preparation, velocity scaling, and state-sensitive timing. Short sequences of actions are prepared collectively in prefrontal cortex, then queued for performance by a cyclic competitive process that operates on a parallel analog representation. Successful acts like ball-catching depend on coordinated scaling of effector velocities, and velocity scaling, mediated by the basal ganglia, may be coupled to perceived time-to-contact. Making acts accurate at high speeds requires state-sensitive and precisely timed activations of muscle forces in patterns that accelerate and decelerate the effectors. The cerebellum may provide a maximally efficient representational basis for learning to generate such timed activation patterns. PMID- 15350245 TI - The Gordian knot of developing tocolytics. PMID- 15350246 TI - Placental oxidative stress: from miscarriage to preeclampsia. AB - OBJECTIVE: To review the role of oxidative stress in two common placental-related disorders of pregnancy, miscarriage and preeclampsia. METHODS: Review of published literature. RESULTS: Miscarriage and preeclampsia manifest at contrasting stages of pregnancy, yet both have their roots in deficient trophoblast invasion during early gestation. Early after implantation, endovascular trophoblast cells migrate down the lumens of spiral arteries, and are associated with their physiological conversion into flaccid conduits. Initially these cells occlude the arteries, limiting maternal blood flow into the placenta. The embryo therefore develops in a low oxygen environment, protecting differentiating cells from damaging free radicals. Once embryogenesis is complete, the maternal intervillous circulation becomes fully established, and intraplacental oxygen concentration rises threefold. Onset of the circulation is normally a progressive periphery-center phenomenon, and high levels of oxidative stress in the periphery may induce formation of the chorion laeve. If trophoblast invasion is severely impaired, plugging of the spiral arteries is incomplete, and onset of the maternal intervillous circulation is premature and widespread throughout the placenta. Syncytiotrophoblastic oxidative damage is extensive, and likely a major contributory factor to miscarriage. Between these two extremes will be found differing degrees of trophoblast invasion compatible with ongoing pregnancy but resulting in deficient conversion of the spiral arteries and an ischemia-reperfusion-type phenomenon. Placental perfusion will be impaired to a greater or lesser extent, generating commensurate placental oxidative stress that is a major contributory factor to preeclampsia. CONCLUSION: Miscarriage, missed miscarriage, and early- and late-onset preeclampsia represent a spectrum of disorders secondary to deficient trophoblast invasion. PMID- 15350247 TI - Apoptosis in the trophoblast--role of apoptosis in placental morphogenesis. AB - Villous trophoblast is the epithelial cover of the placental villous tree and comes in direct contact with maternal blood. The turnover of villous trophoblast includes proliferation and differentiation of cytotrophoblast, syncytial fusion of cytotrophoblast with the overlying syncytiotrophoblast, differentiation in the syncytiotrophoblast, and finally extrusion of apoptotic material into the maternal circulation. In recent years, it has become clear that apoptosis is a normal constituent of trophoblast turnover and the release of apoptotic material does not lead to an inflammatory response of the mother. During preeclampsia there seems to be an altered balance between proliferation and apoptosis of villous trophoblast leading to a dysregulation of the release from the syncytiotrophoblast. The normal apoptotic release may be reduced in favor of a necrotic release. Since apoptosis is still ongoing in the syncytiotrophoblast, a necrotic release of intrasyncytial and partly apoptotic material lead us to call this type of release "aponecrotic shedding." In this situation, cell-free components such as G-actin and DNA freely floating in maternal blood may trigger damage to the maternal endothelium, thereby triggering preeclampsia. This review highlights the importance of the apoptosis cascade in permitting normal physiologic turnover of villous trophoblast. It will demonstrate the participation of initial stages of this cascade within the cytotrophoblast and of the execution stages within the syncytiotrophoblast. Moreover, this review presents hypotheses of how dysregulation of the apoptosis cascade may be linked to endothelial dysfunction of the maternal vasculature in preeclampsia. PMID- 15350248 TI - Amniotic fluid volume responses to amnio-infusion of amniotic fluid versus lactated Ringer's solution in fetal sheep. AB - OBJECTIVE: The present study tested the hypothesis that an intra-amniotic infusion of amniotic fluid (AF) would produce a more sustained increase in AF volume than an infusion of lactated Ringer's solution. METHODS: Five chronically catheterized, late-gestation fetal sheep were studied over two 5-day periods with AF volume measured daily. After baseline measurements on day 1, 1 L of either warmed, previously frozen AF or warmed lactated Ringer's solution was infused intra-amniotically over 60 minutes. Two days later, the other fluid was infused. During the second week, fluids were infused in the opposite order. Analysis of variance (ANOVA) was used for statistical testing. RESULTS: Following intra amniotic infusion (n = 20) of 1007 +/- 7 (SE) mL of either AF or Ringer's solution, intra-amniotic retention of the infused fluid was only moderate after 1 day (37.2% +/- 7.9%, P <.001) and was not significantly different from zero after 2 days (16.5% +/- 9.5%, P =.1). There were no significant differences in AF volume following infusion of AF versus lactated Ringer's solution or the order in which they were infused. AF compositional changes were similar except that pH and bicarbonate concentration were reduced as expected immediately after lactated Ringer's solution with a return to normal values after 1 day. AF lactate increased after lactated Ringer's solution infusion, declining to baseline values after 2 days. Fetal urine flow rate increased by 75% +/- 24% at 1 day postinfusion and there was no difference between infusates. CONCLUSIONS: The expansion of AF volume over 2 days following amnio-infusion does not appear to depend on minor compositional differences or the presence of microconstituents such as hormones, cytokines, or growth factors that are normally present in AF. PMID- 15350249 TI - Effects of umbilical cord occlusion in late gestation on the ovine fetal brain and retina. AB - OBJECTIVE: Previous studies on the effects of umbilical cord occlusion (UCO) on the fetal brain have focused on short-term alterations, and in most cases have used only subjective techniques to evaluate brain injury. Our aim was to assess quantitatively the persistent consequences of UCO on the developing brain; we also examined the retina. METHODS: We subjected fetal sheep to a single episode of UCO at 126 days of gestation (term approximately 147 days) to induce at least 10 minutes of isoelectric fetal electrocorticogram (ECoG). RESULTS: UCO resulted in fetal asphyxia and transient mild alterations in fetal mean arterial pressure (MAP). UCO did not result in significant injury to the developing brain or retina when assessed 15 days after the insult. There was no change in the endogenous expression of brain-derived neurotrophic factor (BDNF) protein in the hippocampus, nor was there a significant loss of CA1 hippocampal pyramidal cells. However, this insult did result in subtle neuropathologic alterations in the brain, including a reduction in the weight of the cerebral hemispheres, an increase in the areal density of cerebellar Purkinje cells, and enlarged perivascular spaces around blood vessels and inflammatory cells in the cerebral white matter. UCO did not affect the thickness of the central or peripheral retina or the numbers of retinal dopaminergic, cholinergic, and nitrergic amacrine cells. CONCLUSIONS: Thus, while 10 minutes of UCO did not result in overt injury to the fetal brain or retina, the observed changes in the fetal brain suggest altered growth of neural processes, which may contribute to neurologic deficits postnatally or to increased vulnerability of the brain to later insults during either the remainder of gestation or after birth. PMID- 15350250 TI - Pharmacokinetics of low molecular weight heparin and unfractionated heparin in pregnancy. AB - OBJECTIVE: Little pharmacokinetic data are available for either low molecular weight heparins (LMWHs) or unfractionated heparins (UFHs) in pregnancy. The objectives of this study were to determine whether differences exist in the pharmacokinetics of dalteparin and UFH before and during the first, second, and third trimesters of pregnancy in women with the antiphospholipid antibody syndrome (APS). Adjustments in our dosing protocol would be made if differences existed. METHODS: Women with APS who were contemplating pregnancy were randomized to dalteparin 2500 U, 2500 U, 5000 U, and 7500 U daily, or UFH 5000 U, 5000 U, 7500 U, and 10,000 U every 12 hours, prior to pregnancy and the first, second, and third trimesters, respectively. Serial plasma concentrations of heparin were measured during 4 blood sampling days by determining anti-factor Xa activity. RESULTS: Fifteen (n = 9 receiving dalteparin and n = 6 receiving UFH) completed all four sampling periods. For dalteparin, significant differences (P <.05) were detected, using area under the curve (AUC), between pre-pregnancy versus third trimester, first versus second trimester, first versus third trimester, and second versus third trimester. No significant differences were detected in the UFH group. CONCLUSION: In APS, our original dosing protocol of dalteparin yielded significant differences (P <.05) in drug exposure throughout pregnancy. Based on these results, we recommend a prophylactic dalteparin dosing regimen of 2500 U every 24 hours pre-pregnancy (and for 6 weeks postpartum), and 5000 U every 24 hours during the first, second, and third trimesters. Due to lack of significant differences in AUC throughout pregnancy for UFH, we recommend continuing with our original dosing protocol. PMID- 15350251 TI - Inhibitory effect of barusiban and atosiban on oxytocin-induced contractions of myometrium from preterm and term pregnant women. AB - BACKGROUND: A synthetic oxytocin analogue, barusiban, was shown to potently inhibit oxytocin-induced activity of myometrium from term pregnant women. The responsiveness to vasopressin was not influenced by the compound. OBJECTIVE: To test the effect of barusiban and a reference compound, atosiban, on oxytocin induced activity of myometrium from women at preterm pregnancy in comparison to myometrium from women at term. METHODS: Fifteen preterm (30-36 gestational weeks) and 12 term pregnant women (38-41 weeks) who underwent cesarean delivery donated myometrial tissue for the study. Concentration-response curves following oxytocin administration to isolated myometrial strips were recorded in control experiments, in the presence of barusiban at concentrations of 2.5, 25, and 250 nM, and of atosiban at concentrations of 25, 250, and 750 nM. Effective concentration 50% (EC50) and pA2 values were calculated. RESULTS: Both antagonists in higher concentrations increased the EC50 values to oxytocin. The median pA2 value for preterm myometrium with barusiban was 9.76 and with atosiban 7.86. For term myometrium the corresponding pA2 results were 9.89 and 7.81, respectively. None of these pA2 values differed to any statistically significant degree. CONCLUSION: The selective oxytocin antagonist, barusiban, concentration dependently inhibits oxytocin-induced myometrial contractions of both preterm and term myometrium at least as potently as atosiban. It remains to be determined if the selectivity of barusiban for the oxytocin receptor confers an advantage over atosiban as a tocolytic in preterm labor. PMID- 15350252 TI - Uterine blood flow response to alpha-adrenergic blockade in nulligravid women of reproductive age. AB - OBJECTIVE: Elevated uterine artery resistance has been associated with the development of preeclampsia. We sought to determine if the human uterine arterial circulation was more sensitive to alpha-adrenergic blockade than a control vascular bed. METHODS: We studied 38 healthy nulliparous women during the follicular phase of the menstrual cycle. Arteriolized venous blood was measured for plasma epinephrine and norepinephrine. Doppler ultrasound of the uterine and upper extremity radial artery was performed before and after intravenous administration of phentolamine. Heart rate and volumetric flow were calculated. Data are presented as means +/- SD. RESULTS: Mean heart rate before and after administration of phentolamine was 63 +/- 11 and 68 +/- 12 beats per minute (7.5% increase, P <.001). Mean radial artery blood flow before and after administration of phentolamine was 4.6 +/- 4.7 mL/min and 5.8 +/- 5.9 mL/min (19.9% increase, P =.071). Mean uterine blood flow before and administration of phentolamine was 15.4 +/- 10.1 mL/min and 27.3 +/- 17.5 mL/min (43.7% increase, P <.001). The uterine response (43.7% increase) and radial response (19.9% increase) were significantly different (P <.001). The increase in uterine blood flow after phentolamine administration was linked with resting norepinephrine (r = 0.394, P =.063). CONCLUSION: We found evidence that uterine alpha-adrenergic tone is more sensitive to blockade by phentolamine than upper extremity radial circulation. This provides evidence for a differential responsiveness and sensitivity to alpha adrenergic blockade in different vascular beds. We theorize that the increased adrenergic tone associated with preeclampsia may contribute disproportionately to decreased uterine blood flow. PMID- 15350253 TI - Dependence of 3',5'-cyclic adenosine monophosphate--stimulated gonadotropin releasing hormone release on intracellular calcium levels and L-type calcium channels in superfused GT1-7 neurons. AB - OBJECTIVE: Immortalized GT1-7 neurons were used to characterize the interactive roles of adenylate cyclase-3',5'-cyclic adenosine monophosphate (cAMP) and L-type calcium channels on gonadotropin-releasing hormone (GnRH) release. METHODS: Dibutyryl (db)-cAMP was used as an active analog of endogenous cAMP, and forskolin was used to activate adenylate cyclase. Extracellular calcium was chelated using EGTA and L-type calcium channels were blocked using nimodipine. The selective Ca2+ ionophore A23187 was employed to increase intracellular calcium levels. GT1-7 neurons were grown on Cytodex-3 beads (Pharmacia Biotech, Uppsala, Sweden) and placed in special superfusion microchambers. The cells were superfused at a rate of 6.2 mL/h with media 199 (M-199; Gibco, Grand Island, NY; pH 7.35, 37C); effluent fractions were collected at 5-minute intervals for analysis of GnRH concentrations by radioimmunoassay. RESULTS: Basal GnRH release from superfused GT1-7 neurons ranged from 10 to 62 pg. min(-1). mL(-1). Coexposure of the cells to forskolin and A23187 produced an additive effect on stimulated release of GnRH. Cells exposed to 1 microM of forskolin (an activator of adenylate cyclase) for 5 minutes showed a 2.6-fold increase in GnRH release. Likewise, the addition of 100 microM of db-cAMP to the superfusion for 5 minutes demonstrated a 2.3-fold increase in the amplitude of GnRH secretion. Maintaining the superfused cells in medium containing 5 mM EGTA had no obvious effect on basal GnRH release but blocked the effect of db-cAMP to increase GnRH release. Similarly, the addition of 10 microM nimodipine to the superfusion medium blocked db-cAMP-stimulated GnRH release. CONCLUSIONS: These findings provide additional evidence that cAMP-mediated GnRH release from GT1-7 neurons is dependent on influx of extracellular calcium via L-type Ca2+ channels. PMID- 15350254 TI - Induction of insulin-like growth factor binding protein-1 expression in baboon endometrial stromal cells by cells of trophoblast origin. AB - OBJECTIVE: To determine the influence of the conceptus on the induction of decidualization in endometrial stromal cells from the baboon. METHODS: For in vivo studies, implantation sites from day 22 of pregnancy in the baboon were analyzed for insulin-like growth factor-II (IGF-II) and insulin-like growth factor binding protein-1 (IGFBP-1) mRNAs using in situ hybridization. For in vitro studies, Jeg-3 cells or primary cytotrophoblasts isolated from term placenta were cocultured with a monolayer of stromal cells from the baboon endometrium. Cytotrophoblasts were placed either directly on top of the stromal cell monolayer or in transwell cell culture inserts and cultured for 7 days. No exogenous hormones were added. Stromal cells were analyzed for IGFBP-1 mRNA and protein by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry. RESULTS: Examination of early implantation sites in the baboon revealed a high expression of IGF-II mRNA in the invading cytotrophoblasts. Conversely, the stromal cells of the endometrium directly adjacent to the cytotrophoblasts expressed IGFBP-1 mRNA. Endometrial stromal cells cocultured with Jeg-3 cells or primary cytotrophoblasts for 7 days expressed IGFBP-1 mRNA and protein. This expression occurred in both the direct coculture system and coculture using the cell culture inserts. Cytotrophoblasts also induced the expression of prolactin in stromal fibroblasts following coculture. CONCLUSIONS: The conceptus is capable of inducing decidualization of endometrial stromal cells. This is shown in both the in vivo and in vitro systems. PMID- 15350255 TI - Aromatase inhibition reduces the dose of gonadotropin required for controlled ovarian hyperstimulation. AB - OBJECTIVE: To compare the use of the aromatase inhibitor, letrozole, in conjunction with follicle-stimulating hormone (FSH) injection, and FSH alone for controlled ovarian hyperstimulation (COH) in patients with polycystic ovarian syndrome (PCOS) or ovulatory infertility. METHODS: This nonrandomized study included two study groups: 26 patients with PCOS and 63 with ovulatory infertility (unexplained infertility [41 patients], male factor infertility [17 patients], and endometriosis [5 patients]), who received letrozole in addition to FSH; and two control groups: 46 PCOS patients and 308 with ovulatory infertility (unexplained infertility [250 patients], male factor infertility [42 patients], and endometriosis [16 patients], who received FSH only. All patients had intrauterine insemination (IUI). Main outcome measures included dose of FSH used per cycle, number of preovulatory follicles greater than 16 mm in diameter, cancellation rate, and pregnancy rate. RESULTS: The FSH dose required for ovarian stimulation was significantly lower when letrozole was used in both study groups compared to the control groups without a significant difference in number of follicles greater than 16 mm. IUI cancellation rate was significantly lower with letrozole treatment in PCOS patients. In women with PCOS, clinical pregnancy rate per completed IUI cycle was 26.5% in the letrozole plus FSH group versus 18.5% in the FSH-only group. In ovulatory infertility patients, the pregnancy rate was similar in both study and control groups (11%). CONCLUSION: We believe that inhibition of estrogen synthesis by aromatase inhibition will release the estrogenic negative feedback, resulting in an increase in endogenous FSH secretion. Moreover, by inhibiting conversion of androgens into estrogens, accumulating androgens may increase follicular sensitivity to FSH. Such a protocol has the potential to lower FSH treatment cost and may improve response for low responders who require high FSH doses during ovarian stimulation. PMID- 15350256 TI - Angiotensin II sensitivity in nonpregnant formerly preeclamptic women and healthy parous controls. AB - BACKGROUND: In women prone to develop hypertensive complications, vascular reactivity fails to decrease in early pregnancy. Since hypertensive syndromes of pregnancy seem to be superimposed on a preexisting disorder, we tested the hypothesis that in formerly preeclamptic women, as compared to healthy parous controls, circulatory reactivity to angiotensin II is enhanced in the follicular phase of the menstrual cycle. METHODS: Sixty formerly preeclamptic women were subdivided into a hypertensive (HYPERT, n = 14), a normotensive thrombophilic (THROMB, n = 26), and a normotensive nonthrombophilic (ASYMPT, n = 20) subgroup. In these women and in 11 healthy parous controls we assessed at least 5 months postpartum at day 5 (+/-2) of the menstrual cycle the following variables: body weight, height, plasma volume, reactivity to infused angiotensin II of arterial blood pressure, heart rate, glomerular filtration rate (GFR), effective renal blood flow, and the hormones of the renin-angiotensin-aldosterone (RAAS) axis. RESULTS: At baseline, THROMB did not differ from controls. In contrast, ASYMPT exhibited slight overweight, reduced plasma volume, and reduced renal blood flow. HYPERT much resembled ASYMPT except for the overweight, renal perfusion, and GFR. Infusion of angiotensin II led to comparable decreases in renal perfusion and filtration, and to increases in blood pressure. However, sensitivity to this substance correlated inversely with relative and absolute sizes of the plasma volume compartment. CONCLUSION: Circulatory sensitivity to infused angiotensin II is comparable between nonpregnant formerly preeclamptic women and healthy parous controls. However, responsiveness to this agent is enhanced among women with a contracted plasma volume compartment, a condition commonly observed among formerly preeclamptic women. PMID- 15350257 TI - General introduction to vasopressin and oxytocin: structure/metabolism, evolutionary aspects, neural pathway/receptor distribution, and functional aspects relevant to memory processing. PMID- 15350258 TI - De Wied and colleagues I: evidence for a VP and an OT influence on MP: launching the "VP/OT central memory theory". PMID- 15350259 TI - De Wied and colleagues II: further clarification of the roles of vasopressin and oxytocin in memory processing. PMID- 15350260 TI - De Wied and colleagues III: brain sites and transmitter systems involved in the vasopressin and oxytocin influence on memory processing. PMID- 15350261 TI - De Wied and colleagues IV: research into mechanisms of action by which vasopressin and oxytocin influence memory processing. PMID- 15350262 TI - Research studies of Koob and colleagues: the "vasopressin dual action theory". PMID- 15350263 TI - Contributions of Sahgal and colleagues: the "vasopression central arousal theory". PMID- 15350264 TI - Role of attentional processing in mediating the influence of vasopressin on memory processing. PMID- 15350265 TI - Expansion of vasopressin/oxytocin memory research I: peripheral administration. PMID- 15350266 TI - Expansion of vasopressin/oxytocin memory research II: brain structures and transmitter systems involved in the influence of vasopressin and oxytocin on memory processing. PMID- 15350267 TI - Expansion of vasopressin/oxytocin memory research III: research summary and commentary on theoretical and methodological issues. PMID- 15350268 TI - Research contributions of Dantzer, Bluthe, and colleagues to the study of the role of vasopressin in olfactory-based social recognition memory. PMID- 15350269 TI - Expansion of olfactory-based social recognition memory research: the roles of vasopressin and oxytocin in social recognition memory. PMID- 15350270 TI - Brain-fluid barriers: relevance for theoretical controversies regarding vasopressin and oxytocin memory research. PMID- 15350271 TI - Closing remarks: review and commentary on selected aspects of the roles of vasopressin and oxytocin in memory processing. PMID- 15350274 TI - Early gene expression in the organ of Corti exposed to gentamicin. AB - Studies have demonstrated different pathogenetic key factors in gentamicin induced hair cell death. The production of reactive oxygen species (ROS), as well as apoptosis-related genes, play a critical role. However, a coordinated large scale investigation of gene expression in the organ of Corti (OC) exposed to gentamicin has not yet been conducted. Here we used DNA microarray technology to compare the expression profile of OC exposed to gentamicin to the expression profile of untreated OC. The OCs of Sprague Dawley rats were dissected and the basal turns were cultured. Two-thirds of the explants were then exposed to l00 microM gentamicin, for 4 and 8 h, while one-third of the explants remained in culture medium alone. Gene expression was analyzed using DNA microarray technology and the dChip software package. Based on the results, the 4-h time point was chosen for further analysis. In these assays, out of 8800 genes, 12 genes were identified on the basis of differential expression in the OC exposed to gentamicin vs. control OC. The identity of these genes suggests that the response of the OC to the gentamicin challenge involves down-regulation of specific gene families in order to alleviate ROS and N-methyl-D-aspartate (NMDA) receptor-mediated cellular stress. PMID- 15350275 TI - The immunohistochemical analysis of pendrin in the mouse inner ear. AB - Pendred's syndrome (PS) is an autosomal recessive disorder characterized by deafness and goiter, which are caused by mutations in the Pendred's syndrome gene (PDS). PDS encodes a membrane protein named pendrin that is considered to act as an anion transporter. An expression pattern of the PDS ortholog (Pds) mRNA in the auditory and vestibular systems has been reported in mice, and the localization of pendrin has been reported recently. We generated antipeptide antibodies against human pendrin, and performed immunohistochemical analysis of mouse inner ears. We detected pendrin in the endolymphatic duct and sac, and the utricle, saccule, and external sulcus. In the endolymphatic duct and sac, the expression of pendrin was apparent at the apical membrane. In addition, we detected pendrin in the spiral ligament, Claudius cells, Deiter's cells, and the spiral ganglion of the cochlea. Our results are key to defining the role of pendrin in inner ear development and elucidating the pathogenic mechanisms underlying deafness in PS. PMID- 15350276 TI - Evidence for tinnitus-related plasticity in the auditory and limbic system, demonstrated by arg3.1 and c-fos immunocytochemistry. AB - Distributions of arg3.1 and c-fos immunoreactive neurons (IRN) in gerbil auditory cortex (AC) and amygdala showed characteristic differences when comparing systemic application of the tinnitus-eliciting drug salicylate with acoustic stimulation or saline injections. In AC, arg3.1 IRN induced by stimulation focused in regions corresponding to the frequency content of the stimulus. Injections of salicylate (350 mg/kg body weight) led to accumulation of arg3.1 IRN in the high frequency domain, while saline injections produced a diffuse distribution. After all treatments, c-fos IRN outnumbered arg3.1 IRN in AC and showed a broad distribution. In subcortical auditory structures arg3.1 IRN were absent in all but one brain. In ventral cochlear nucleus, c-fos IRN were always found after stimulation and often also after saline injections, whereas none were present when injecting salicylate. Similarly, in inferior colliculus, numbers of c-fos IRN were lowest after salicylate injections. In the amygdala, c-fos and arg3.1 IRN were increased substantially after salicylate injections compared to auditory stimulation or saline injections. In particular in its central nucleus, c-fos and arg3.1 IRN were found exclusively after the tinnitus-inducing treatment, suggesting that coactivation of the AC and the amygdala may by an essential feature of tinnitus-related activation. PMID- 15350277 TI - A novel RFP-RET transgenic mouse model with abundant eumelanin in the cochlea. AB - We report on the cochlea of a novel metallothionein-I (MT)/RFP-RET transgenic mouse model with severe systemic melanosis. Electron microscopy revealed that these transgenic mice possess abundant quantities of melanin in the intermediate cells of the stria vascularis. High performance liquid chromatography analysis indicated that cochleae of these transgenic mice contained about twice as much eumelanin as cochleae of control C57BL/6 mice and that the amount of pheomelanin was approximately equal in these two strains. Auditory brainstem responses at 2, 4, 8, and 16 kHz were not significantly different between transgenic and control mice. This is the first report on a mouse model of overproduction of cochlear eumelanin, and our results suggest that this transgenic mouse is an excellent model for investigating the effects of overexpression of cochlear eumelanin. In addition, we provide evidence that eumelanin overproduction in the cochlea does not affect normal hearing. PMID- 15350278 TI - The effect of hair bundle shape on hair bundle hydrodynamics of non-mammalian inner ear hair cells for the full frequency range. AB - The effect of the size and the shape of the hair bundle of a hair cell in the inner ear of non-mammals on its motion for the full range of frequencies is determined thereby extending the results of a previous analysis of hair bundle motion for high and low frequencies [Hear Res. 141 (2000) 39-50]. A hemispheroid is used to represent the hair bundle because it can represent a full range of shapes, from thin, pencil-like shapes to wide, flat, disk-like shapes. Boundary element methods are used to approximate the solution for the hydrodynamics. For physiologically relevant parameters, an excellent match is obtained between the model's predictions and measurements of hair bundle motion in the free-standing region of the basilar papilla of the alligator lizard [Aranyosi, Measuring sound induced motions of the alligator lizard cochlea. Massachusetts Institute of Technology, PhD Thesis, 2002]. Neither in the model's predictions nor in experimental measurements is sharp tuning observed. The model predicted the low frequency region of neural tuning curves for the alligator lizard and bobtail lizard, but could not predict the sharp tuning or the high frequency region. An element that represents an active mechanism is added to the hair bundle model to predict neural tuning curves, which are sharply tuned, and an excellent match is obtained for all the characteristics of neural tuning curves for the alligator lizard, and for the low and high frequency regions for the bobtail lizard. The model does not predict well the sharp tuning of the shorter hair bundles of the bobtail lizard, possibly because it does not represent tectorial sallets. PMID- 15350279 TI - Effects of selected pharmacological agents on avian auditory and vestibular compound action potentials. AB - Glutamate is currently the consensus candidate for the hair cell transmitter in the inner ear of vertebrates. However, other candidate transmitter systems have been proposed and there may be differences in this regard for auditory and vestibular neuroepithelia. In the present study, perilymphatic perfusion was used to deliver prescribed concentrations of ten drugs to the interstitial fluids of the inner ear of hatchling chickens (n = 124). Dose-response curves were obtained for four of these pharmacological agents. The work was carried out in part to distinguish further the neuroepithelial chemical receptors mediating auditory and vestibular compound action potentials (CAPs). Kainic acid (KA) eliminated both auditory and vestibular responses. D-alpha-Aminoadipic acid (DAA) and dizocilpine maleate (MK-801), both NMDA-specific antagonists, failed to alter vestibular CAPs at any concentration. MK-801 significantly and selectively reduced auditory CAPs at concentrations equal to or greater than 1 mM. Similarly, kynurenic acid (4 hydroxyquinoline-2-carboxylic acid, 1 mM), a glutamate antagonist, significantly reduced auditory but not vestibular CAPs. A non-NMDA glutamate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), reduced vestibular CAPs significantly but only at the highest concentration tested (1 mM). In contrast, CNQX reduced auditory responses at concentration as low as 1 microM. The CNQX concentration effective in reducing auditory CAPs by 50% (EC(50)) was approximately 20 microM. Glutamate (1 mM) as well as alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid (AMPA), a glutamate agonist, significantly reduced auditory CAPs (AMPA EC(50)=100 microM). Bicuculline, a GABA(A) receptor antagonist, and L-NAME, a nitric oxide synthase inhibitor, failed to alter responses from either modality. These findings support the hypothesis that glutamate receptors mediate auditory CAPs in birds. However, the results underscore a remarkable difference in sensitivity of the vestibular neuroepithelium (here gravity receptors) to non-NMDA receptor antagonists. The basis of the vestibular insensitivity to glutamate blockers is unknown but it may reflect differences in receptors themselves, differences in the transmission modes available to vestibular synapses or differences in the access of compounds to vestibular neuroepithelial receptors from the interstitial-perilymphatic fluid spaces. PMID- 15350280 TI - Intracellular Ca2+ buffers can dramatically affect Ca2+ conductances in hair cells. AB - The effects of endogenous and exogenous Ca(2+) buffers on Ca(2+) current kinetics have been investigated by patch clamp in hair cells mechanically isolated from frog semicircular canals. This preparation displays at least three different Ca(2+) channel types: transient currents flow through a drug-resistant channel ("R1"), while non-inactivating channels sustain a steady, plateau current comprised of a large L component and a small drug-resistant fraction ("R2"). In the perforated-patch condition a large and stable Ca(2+) current was recorded, with all three components. In whole-cell, a buffer-free pipette solution did not prevent a complete Ca(2+) response. The size of the transient and plateau current fractions were greatly reduced, but the ratio between the two fractions, as well as the activation, inactivation and deactivation kinetics, were substantially unmodified. Current amplitude partially recovered with 5 mM EGTA in the pipette solution. With 50 mM EGTA all the kinetic parameters were slowed down and the transient component, but not the plateau component, markedly increased in size. Response kinetics slowed down even more with 30 mM Cs-BAPTA and the Ca(2+) waveform was substantially modified. The transient component was very large and inactivated slowly; the remaining very small plateau fraction deactivated along a slow, single exponential time. Under this condition nifedipine (10 microM) produced a great reduction of the transient current, leaving plateau and deactivation phase unaltered. This suggests that only R2 channels were still active at the end of the test and that the minor remaining transient component flowed through slowly but completely inactivating R1 channels. These results confirm the presence of several channel types in semicircular canal receptors, at difference with cochlear hair cells, and highlight a dramatic alteration of L type channel behavior when intracellular Ca(2+) buffers are sufficiently concentrated and fast to interfere with rapid and local changes in Ca(2+) levels. PMID- 15350281 TI - Speech offsets activate the right parietal cortex. AB - Speech offsets, i.e., sudden transitions from continuous speech sound to silence, activated both hemispheres differently. In addition to peak activities in the bilateral temporal cortices at about 120 ms after the offsets, the right parietal cortex was activated later irrespective of the stimulated ear. The result was discussed in the context of auditory attention. PMID- 15350282 TI - Contributions of age, cochlear integrity, and auditory environment to avian cochlear nucleus metabolism. AB - Most commercially raised broiler chickens display progressive cochlear degeneration with age [Hear. Res. 166 (2002) 82]. Recent work examining the effects of age and cochlear degeneration on avian cochlear nucleus (nucleus magnocellularis, NM) metabolism showed that changes in metabolic activity occur with age and cochlear damage [Hear. Res. 175 (2003) 101]. The auditory environment also differed between facilities housing young and adult birds. The relative contributions of age, cochlear degeneration, and auditory environment to these changes in NM metabolism are unknown. Using cytochrome oxidase (CO) histochemistry, NM neuron metabolism is examined in several age groups of birds under varying conditions. When normal cochlear integrity and auditory environment are held constant, CO staining is significantly decreased in adult vs. young birds. When age and auditory environment are held constant, CO staining is significantly decreased in birds with damaged vs. normal cochleae. When age and normal cochlear integrity are held constant, CO staining is significantly decreased in birds living in a quiet vs. noisy environment. All factors examined cause changes in CO staining, which is indicative of NM metabolic activity. Results are discussed in the context of mitochondrial aging, afferent regulation, and auditory deprivation and enrichment. PMID- 15350283 TI - Haplo-insufficiency revealed in deafwaddler mice when tested for hearing loss and ataxia. AB - The auditory and vestibular systems rely on the plasma membrane calcium ATPase, isoform 2 (PMCA2) to extrude calcium that enters the stereocilia during transduction. Mutations in the gene encoding this protein result in recessive sensorineural deafness and ataxia in the deafwaddler mouse. In this study, we report the identification of a new allele of deafwaddler, dfw(3j). This allele contains a 4-nucleotide deletion resulting in a frame-shift and predicted truncation of PMCA2. No protein is detected in dfw(3j) homozygotes. To examine the dependence of auditory and vestibular function on PMCA2 activity, we compared dfw(3j) with another functional null allele, dfw(2j), and the partial loss-of function allele, dfw. All mice studied were in the good-hearing CBA/CaJ background. Heterozygotes of either functional null allele displayed highly significant hearing loss by auditory-evoked brainstem responses relative to controls (P < 0.0001), particularly at high frequencies (> 24 kHz). Ataxia was also apparent in these mice on an accelerating rotarod (P < 0.05). In contrast, +/dfw mice were not measurably different from controls in either behavioral test. dfw/dfw mice were deaf, but showed less ataxia than dfw(2j)/dfw(2j) or dfw(3j)/dfw(3j) mice. These results demonstrate that hearing loss and ataxia are dependent on gene dosage and PMCA2 dysfunction. PMID- 15350284 TI - Mechanisms of hearing loss resulting from middle-ear fluid. AB - Fluid in the middle ear, a defining feature of otitis media with effusion (OME), is commonly associated with a 20- to 30-dB conductive hearing loss. The effects and relative importance of various mechanisms leading to conductive hearing loss were investigated in a human temporal bone preparation. Umbo velocity in response to ear-canal sound was measured with a laser vibrometer while saline and silicone fluids of viscosity 5-12,000 cSt were introduced into the middle ear to contact part or all of the tympanic membrane (TM) and fill part or all of the middle ear. At low frequencies, reductions in umbo velocity (deltaVU) of up to 25 dB depended on the percentage of the original middle-ear air space that remained air-filled, which suggests that the primary mechanism in hearing loss at low frequencies is a reduction of the admittance of the middle-ear air space due to displacement of air with fluid. At higher frequencies, deltaVU (of up to 35 dB) depended on the percentage of the TM contacted by fluid, which suggests that the primary mechanism at high frequencies is an increase in tympanic membrane mass by entrained fluid. The viscosity of the fluid had no significant effect on umbo velocity. deltaVU for the fluid-filled middle ear matched hearing losses reported in patients whose middle ear was believed to be completely filled with fluid. The difference between deltaVU for a partly-filled middle ear and hearing losses reported in patients whose middle ear was believed to be incompletely fluid filled is consistent with the reported effect of middle-ear underpressure (commonly seen in OME) on umbo velocity. Small amounts of air in the middle ear are sufficient to facilitate umbo motion at low frequencies. PMID- 15350286 TI - Intelligent therapeutics: biomimetic systems and nanotechnology in drug delivery. PMID- 15350287 TI - Nanotechnology and biomimetic methods in therapeutics: molecular scale control with some help from nature. AB - Nanoscale science and engineering has provided new avenues for engineering materials with macromolecular and even molecular precision. In particular, researchers are beginning to mimic biological systems, achieving molecular scale control via self-assembly and directed assembly techniques. Fabrication and manipulation with macromolecular and molecular precision have led and will lead to the development of novel materials, and these materials will facilitate the fabrication of micro- and nanoscale devices, such as self-regulated micro- and nanoscale drug delivery devices that combine diagnostic and therapeutic actions for instantaneous administration of therapy. As the field of nanoscale science and engineering matures, technologies that will revolutionize the way health care is administered will continue to be developed. PMID- 15350288 TI - Bottom-up design of biomimetic assemblies. AB - Nature has evolved the ability to assemble a variety of molecules into functional architectures that can specifically bind cellular ligands. Mimicking this strategy requires the design of a set of multifaceted molecules, where elements that direct assembly were conjugated to biologically specific components. The development of functional molecular building-blocks that assemble to form compartments for therapeutics addresses the desire to have controllable morphologies that interact with biological interfaces at nanometer length scales. The practical application of such 'bottom-up' assemblies requires the ability to predict the type of aggregated structure and to synthesize molecules in a highly controlled fashion. This bottom-up approach results in a molecular platform that mimics biological systems with potential for encapsulating and delivering drug molecules. PMID- 15350289 TI - BioMEMS: state-of-the-art in detection, opportunities and prospects. AB - In recent years, the biological and biomedical applications of micro- and nanotechnology (commonly referred to as Biomedical or Biological Micro-Electro Mechanical Systems [BioMEMS]) have become increasingly prevalent and have found widespread use in a wide variety of applications such as diagnostics, therapeutics, and tissue engineering. While research and development activity in this field stays intense, some applications have also been commercialized. This article reviews the recent advances in this very exciting and important field and presents a summary of the state of the art in the area of BioMEMS focusing on diagnostics, sensing, and detection. The areas of therapeutics and hybrid bio/artificial devices will be presented in more detail elsewhere [Biomedical Nanotechnology, Vol. I-IV, Maruo Ferrari (Ed.), Kluwer Academic Publishers, 2004, in press.] and here are discussed briefly in terms of future directions and prospects. PMID- 15350290 TI - Biomimetic materials and micropatterned structures using iniferters. AB - In the preparation of biomimetic materials it is often required that efficient methods of polymerization be used, often methods that can lead to biomimetic polymers with relatively narrow molecular weight distribution. Living radical polymerization techniques have successfully been used to create low polydispersity linear polymers by free-radical polymerizations. Although this technique slows down the polymerization of multifunctional monomers, there is little effect on the network structure due to the high concentration of pendent double bonds. There are applications of the living radical polymerization in the synthesis of block copolymers. Essentially, the technique involves polymerizing a single type of monomer first to create a macromonomer that is capable of acting as an initiator because of the reversible bond between the polymer end group and the terminating group. This terminating group may be a thiol or a halogen and, under the right conditions, will dissociate to form radicals. A second monomer is then added to the system and the polymerization proceeds with the second monomer chemically attached to the polymer of the first monomer. We review methods of creating biomimetic block copolymers using the iniferter radical polymerization technique. The block copolymers would be used in the synthesis of micropatterned polymer films for use in biomaterials and other biomedical applications. PMID- 15350291 TI - Configurational biomimesis in drug delivery: molecular imprinting of biologically significant molecules. AB - This review focuses on trends in the macromolecular recognition of biologically significant molecules (e.g., drugs, amino acids, steroids, nucleotide bases, carbohydrates, etc.) via molecular imprinting methods. An extensive list of prior art including type of functional monomers and crosslinkers for each biomolecule imprinted polymer is presented. Representative samples of receptor-ligand dissociation constants and polymer capacities are presented as well as typical values that occur in classes of biological recognition systems. Imprinting technology has direct impact in enhanced drug loading of controlled-release carriers for the sustained release of therapeutic agents as well as robust biosensors for novel therapeutic and diagnostic devices. This review also discusses the future of designed recognition, configurational biomimesis within polymeric gels, and highlights recent efforts toward integrating imprinted polymers in controlled drug delivery systems and sensing devices. In particular, the application of imprinted polymers for sustained release, enhanced loading capacity, and enantioselective loading or release are discussed. This article also highlights the most important problems to be solved in the design of synthetic recognition-based networks for biological molecules. PMID- 15350292 TI - Micro and nano-fabrication of biodegradable polymers for drug delivery. AB - This paper presents state-of-the-art micro and nano-fabrication techniques for biodegradable polymers. Replication molding, using a rigid or elastic master, can pattern structures on a polymer surface in a submicron resolution at a low cost. Layer-by-layer rapid prototyping methods are promising in producing controlled release units with complicated geometries, release mechanisms and the ability to control microstructure and composition. Special attention is paid to the fast, flexible, and non-invasive laser fabrication techniques that have great potential in the fabrication of biodegradable polymer drug delivery devices in both a laboratory and industry scale. PMID- 15350293 TI - Three-dimensional tissue fabrication. AB - In recent years, advances in fabrication technologies have brought a new dimension to the field of tissue engineering. Using manufacturing-based methods and hydrogel chemistries, researchers have been able to fabricate tissue engineering scaffolds with complex 3-D architectures and customized chemistries that mimic the in vivo tissue environment. These techniques may be useful in developing therapies for replacing lost tissue function, as in vitro models of living tissue, and also for further enabling fundamental studies of structure/function relationships in three dimensional contexts. Here, we present an overview of 3-D tissue fabrication techniques based on methods for: scaffold fabrication, cellular assembly, and hybrid hydrogel/cell methods and review their potential utility for tissue engineering. PMID- 15350294 TI - Nanoparticle and targeted systems for cancer therapy. AB - This review explores recent work directed towards more targeted treatment of cancer, whether through more specific anti-cancer agents or through methods of delivery. These areas include delivery by avoiding the reticuloendothelial system, utilizing the enhanced permeability and retention effect and tumor specific targeting. Treatment opportunities using antibody-targeted therapies are summarized. The ability to treat cancer by targeting delivery through angiogenesis is also discussed and antiangiogenic drugs in clinical trials are presented. Delivery methods that specifically use nanoparticles are also highlighted, including both degradable and nondegradable polymers. PMID- 15350296 TI - Nanoscale technology of mucoadhesive interactions. AB - Nanoscale analysis may be used to design new types of mucoadhesive polymers. Understanding of the surface interactions between hydrophilic polymer surfaces and mucins can lead to improved adhesive bonding by hydrogen bonding. Alternatively, decoration of a mucoadhesive polymer surface with tethers of linear and block copolymers containing neutral or ionizable structures provides increased interdigitation and interpenetration with the mucus. Finally, formation of micro- or nanopatterns on these surfaces can lead to promising new systems of oral delivery applications. PMID- 15350295 TI - Nanoporous microsystems for islet cell replacement. AB - The inadequacy of conventional insulin therapy for the treatment of Type I diabetes has stimulated research on several therapeutic alternatives, including insulin pumps and controlled release systems for insulin. One of the most physiological alternatives to insulin injections is the transplantation of insulin-secreting cells. It is the beta cells of the islets that secrete insulin in response to increasing blood glucose concentrations. Ideally, transplantation of such cells (allografts or xenografts) could restore normoglycemia. However, as with most tissue or cellular transplants, the cellular grafts, particularly xenografts, are subjected to immunorejection in the absence of chronic immunosuppression. Thus, it is of great interest to develop new technologies that may be used for islet cell replacement. This research proposal describes a new approach to cellular delivery based on micro- and nanotechnology. Utilizing this approach, nanoporous biocapsules are bulk and surface micromachined to present uniform and well-controlled pore sizes as small as 7 nm, tailored surface chemistries, and precise microarchitectures, in order to provide immunoisolating microenvironments for cells. Such a design may overcome some of the limitations associated with conventional encapsulation and delivery technologies, including chemical instabilities, material degradation or fracture, and broad membrane pore sizes. PMID- 15350297 TI - Challenging pedigrees seen in a hereditary cancer consultation center. AB - The clinical translation of the significance of cancer "running in families" has become a source of major contention as a result of the explosion of knowledge about cancer causality at the molecular level. At the clinical level, the increasing awareness of the familial and hereditary burden of cancer has contributed heavily to both physician and lay concern about cancer risk. Problems in interpretation of the significance of a cancer family history may arise due to a variety of factors, and even if the physician correctly diagnoses a hereditary cancer-predisposing syndrome there may still be barriers to patient compliance with surveillance and management recommendations. Our purpose is to discuss a variety of potential barriers in the diagnosis and management of patients at increased hereditary cancer risk, drawing on examples from a cohort of approximately 300 families evaluated at Creighton University's Hereditary Cancer Consultation Center over 8 years. Each case was selected because of the presence of one or more clinical, pathologic, molecular genetic, psychosocial, economic, confidentiality, or insurance or employment discrimination factors that had the potential to pose an obstacle in diagnosis or in patient compliance with screening and management recommendations. PMID- 15350298 TI - Molecular cytogenetic parameters in fibroblasts of ataxia telangiectasia carrier. AB - Ataxia telangiectasia (AT) is a pleiotropic and rare (1:40,000 to 1:100,000) recessive disease. Laboratory investigations have failed to detect any consistent anomaly in cells from AT heterozygotes. To estimate random aneuploidy, we applied a fluorescence in situ hybridization technique with alpha-satellite probes for chromosomes 8 and 9 and replication pattern for RB-1, HER-2/neu, and the imprinted SNRPN loci on primary AT carrier fibroblasts. Higher random aneuploidy was not found in the carrier fibroblasts compared to control amniocytic cells. The asynchrony pattern was higher in the AT carrier cells with the RB-1 locus (P=0.057) and significantly higher with the HER-2/neu locus (P < 0.001) compared to control cells. As for the imprinted locus SNRPN, there was a significantly lower asynchrony rate in the AT carriers (P < 10(-5)) compared to the control group. Molecular cytogenetic parameters of random aneuploidy and replication pattern may reflect predisposition for the development of cancer. It is possible that in some AT carriers the genetic instability phenomena associated with the abnormal replication pattern may represent their potential for developing malignancies. PMID- 15350300 TI - Cytogenetics and fluorescence in situ hybridization studies of diffuse large B cell lymphoma in children and young adults. AB - Diffuse large B-cell lymphoma (DLBCL), the most common subtype of adult non Hodgkin lymphoma (NHL), is infrequently seen in adolescents and is rare in children. Due to the infrequency of the disease, single institution-based cytogenetic and fluorescence in situ hybridization (FISH) studies of pediatric DLBCL have not been reported so far and, hence, the possible differences in pediatric and adult DLBCL have not been evaluated. We performed cytogenetic and FISH analyses of 7 pediatric and 5 young adult DLBCL cases referred to the University of Nebraska Medical Center. Karyotypic studies revealed numeric and structural chromosome abnormalities in all cases. Loss of chromosomes 2, 3, 4, 6, 12, 15, 16, and 17, and gain of 12, 18, and X were observed in more than 20% of the cases (#10878;3 cases). Sex chromosome abnormalities and cytogenetically unidentifiable chromosomes and/or segments were observed in 80% (10/12) of the cases. Recurrent breakpoints (observed in 3 or more cases) included 14q32 (IGH) and 17p13 (TP53), which clustered in the young adult group. The breakpoints 7q36, 9p24, 13q34, and 16q24 were noted in two cases each. We performed interphase FISH studies to verify the possible rearrangements of the breakpoints that are frequently implicated in adult DLBCL. Our results confirmed that the pediatric cases did not show rearrangements of 3q27 (BCL6), 14q32 (IGH), 18q21 (BCL2), 8q24 (CMYC), and 17p13 (TP53), except for one case with IGH;BCL2 dual fusion [t(14;18)(q32;q21)] and one with a 17p13 (TP53) deletion. Although 3q27 was noted to be rearranged by conventional cytogenetics in two young adult DLBCL cases, FISH investigations verified that BCL6 was not disrupted. The t(8;14)(q24;q32) with rearranged CMYC ascertained by FISH, was observed in a single young adult DLBCL case. These results highlight a distinctly different representation of cytogenetic abnormalities in pediatric versus adult DLBCL. PMID- 15350299 TI - Multiple epithelial and nonepithelial tumors in hereditary nonpolyposis colorectal cancer: characterization of germline and somatic mutations of the MSH2 gene and heterogeneity of replication error phenotypes. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal inherited cancer syndrome characterized by germline plus somatic mutations of DNA mismatch repair genes and familial clustering of cancers of colorectum and other visceral organs. So far, to our knowledge, there has been no proof of nonepithelial tumors in association with HNPCC. Here we report on a MSH2 frameshift HNPCC family with a carrier found to have multiple primary tumors, including endometrial hyperplasia, ovarian adenocarcinoma, skin cavernous hemangioma, and skin dermatofibrosarcoma protuberans (DFSP). We studied the replication error (RER) phenotype in noncoding (Bat-26, Bat-25, D2S123, D5S346, and D17S250) and coding (MSH3, MSH6, BAX, and TGFBR2 genes) DNA sequences, and characterized the germline and somatic mutations of the MSH2 gene in the tumors described above and in endometrial carcinomas from two of her affected siblings. RER was observed in an order of hyperplasic endometrium (6/10 markers), ovarian carcinoma (5/10 markers), endometrial carcinomas (4/9 and 3/10), DFSP (2/9 markers), and cavernous hemangioma (2/10 markers). All the tumors showed the same germline mutation of G5-->G6 frameshift at 183-187 and polymorphism of C1168T in a heterozygous pattern. In an endometrial carcinoma, deletion of the second allele of MSH2 was evident. Heterogeneous RER patterns were noted in multiple primary tumors of the same individual and in premalignant and malignant endometrial tumors from different individuals. The study demonstrated the two hits of the hMSH(2) gene as well as intra- and interindividual variations of RER phenotypes in HNPCC. The first characterized nonepithelial tumors in HNPCC seem to carry a limited panel of RER, including a framesift at the (A)(10) tract of TGFBR2. PMID- 15350301 TI - Polymorphisms in the hypoxia inducible factor-1alpha gene (HIF1A) are associated with the renal cell carcinoma phenotype. AB - Hypoxia inducible factor 1 (HIF-1) is a key regulator of the genes involved in the cellular response to hypoxia. HIF consists of alpha and beta subunits, with the alpha subunit being degraded under normoxic conditions and stabilized under hypoxia. We investigated C1772T and G1790A polymorphisms in exon 12 of the HIF gene, which result in an amino acid change from proline 582 to serine and from alanine 588 to threonine, respectively. These polymorphisms are found within the oxygen-dependent degradation domain of the HIF-1alpha protein and may be important in the oxygen regulation of the protein via hydroxylation of the proline residue at position 564 (P564) by HIF-alpha prolyl hydroxylase (HIF-PH). The frequency of these polymorphisms was studied in 160 nontumor DNA samples from patients with renal cell carcinoma (RCC). There was a highly significant increase in the frequency of both the G/A1790 (45.9 vs. 13.5%, P < 0.00001) and C/C1772 (10 vs. 0.7%, P=0.0004) genotypes in patients with RCC compared with normal healthy controls. A decrease was seen for the GG (44.5 vs. 83%, P < 0.00001) and CT (33.8 vs. 55.5%, P=0.0001) genotypes in patients compared with controls. There was a marked increase in the T-A haplotype (22.8 vs. 9.5%, P=0.00008) and an increase in the C-A haplotype (4.9 vs. 1.1%, P=0.02) in patients compared with controls, and a decrease in the T-G haplotype (53.4 vs. 65.1%, P=0.01). No statistical difference was found for the other haplotypes. These findings show that polymorphisms of the HIF1A gene may confer susceptibility to RCC. PMID- 15350302 TI - Aneuploidies, deletion, and overexpression of TP53 gene in intestinal metaplasia of patients without gastric cancer. AB - Gastric carcinogenesis is attributable to interacting environmental and genetic factors, through a sequence of events including intestinal metaplasia. Using a fluorescence in situ hybridization technique, we investigated the occurrence of aneuploidies of chromosomes 3, 7, 8, 9, and 17, TP53 gene deletion, and expression of p53 in 21 intestinal metaplasia (IM) samples from cancer-free patients and in 20 gastric adenocarcinoma samples. Aneuploidies were found in 71% (15/21) of the IM samples. Trisomy of chromosomes 7 and 9 occurred mainly in complete-type IM; in the incomplete type, trisomy of chromosomes 7 and 8 were more commonly found. The TP53 gene deletion was observed in 60% (3/5) of the IM cases, and immunohistochemistry revealed p53 overexpression in 12% (2/17) of the analyzed IM cases. All gastric adenocarcinoma cases presented higher frequencies of trisomy or tetrasomy of chromosomes 3, 7, 8, 9, and 17. The TP53 deletion was found in all three of the gastric adenocarcinoma analyzed for it, and immunohistochemistry detected overexpression of protein p53 in 80% (12/15) of the analyzed cases. Our study revealed for the first time the presence of aneuploidies of chromosomes 7, 8, 9, and 17 and of TP53 gene deletion and overexpression in IM samples from cancer-free patients. These results suggest that IM and gastric adenocarcinoma may share the same genetic alterations. PMID- 15350303 TI - Effect of interferon-alpha on chromosome abnormalities in treated chronic myelogenous leukemia patients. AB - To investigate the relationship of chromosomal aberrations at blastic crisis (BC) in chronic myelogenous leukemia (CML), with previous therapies and with atomic bomb (AB) exposure, we studied 114 CML patients who developed BC, including 23 AB survivors in Hiroshima. In total, only 45.6% showed major-route abnormalities, which figure was far lower than those previously reported, implying possibility of geographical difference. Occurrence of major-route abnormality was not associated with either duration of chronic phase or survival time after BC. Patients treated with interferon-alpha (IFNalpha) showed lower frequency of major route abnormalities and lower number of abnormal chromosomes than did patients treated with busulfan (Bu). The frequency of trisomy 8 was lower and monosomy 7 was higher in IFNalpha-treated than in Bu-treated patients. The frequency of unusual abnormalities at BC in IFNalpha-treated patients was indistinguishable from those in Bu-treated patients and, notably, a more common (40%) feature in IFNalpha-treated patients was no change in the cytogenetic picture. Thus, we conclude that IFNalpha action on chromosome aberration is basically quite neutral and that IFNalpha does not induce any specific aberrations, including unusual ones at BC, with an exception of deletion of chromosome 7. Atomic bomb exposure status did not make any difference in secondary abnormalities at BC. PMID- 15350304 TI - Molecular cytogenetic characterization of the mantle cell lymphoma cell line GRANTA-519. AB - Combining fluorescence R-banding, fluorescence in situ hybridization and spectral karyotyping allowed us to precisely define chromosomal breakpoints, gains, losses and a newly detected amplification in the human mantle cell lymphoma (MCL) cell line GRANTA-519. GRANTA-519 is characterized by the t(11;14)(q13;q32) resulting in overexpression of cyclin D1, a key player in cell cycle control. Hitherto unresolved complex rearrangements involve 1p, 1q, 3cen, 9p, 11q, 12p, 12q, 16p, 17p, and 18cen. Moreover, a 4- to 6-fold gain of sequences on 18q leads to a low level amplification of the BCL2 gene and to an overexpression of the BCL2 protein. These results provide the basis for the identification of not only candidate oncogenes responsible for MCL in gained regions, but also for the identification of putative tumor suppressor genes in commonly deleted regions like 1p22, which would eventually enable functional studies of these genes. PMID- 15350305 TI - Overlapping deletion regions at 11q23 in myelodysplastic syndrome and chronic lymphocytic leukemia, characterized by a novel BAC probe set. AB - Translocations or deletions involving the 11q23 region have been observed in acute lymphoblastic leukemia (ALL), acute myelocytic leukemia (AML), myelodysplastic syndrome (MDS), and chronic lymphocytic leukemia (CLL). BAC probes encompassing the D11S29 and D11S924 markers and flanking the MLL gene were used in dual color fluorescence in situ hybridization. Fifteen patients with hematologic malignancies and cytogenetic abnormalities of 11q23 were analyzed. The BAC and MLL probes demonstrated split signals in five of 7 ALL or AML cases with translocations of 11q23. Of the remaining 2 cases, one had normal signals for both probe sets and the other had a submicroscopic deletion of the MLL 3' region. In one case of AML with del(11)(q23), deletion of the MLL 3' region and the region telomeric to the MLL gene was seen. Three CLL cases with deletion of part or the entire 11q23 region showed deletion of one copy of MLL, but retention of the region telomeric to MLL. However, in four MDS cases with deletions involving the 11q23 region, deletions of both the MLL gene and the flanking regions of the MLL gene were observed. Hence, the deletions on 11q23 are different but overlapping for CLL and MDS, implicating different genes involved for these diseases. PMID- 15350306 TI - Combined spectral karyotyping, multicolor banding, and microarray comparative genomic hybridization analysis provides a detailed characterization of complex structural chromosomal rearrangements associated with gene amplification in the osteosarcoma cell line MG-63. AB - The advancement of fluorescence in situ hybridization-based assays has permitted more refined delineation of chromosomal loci involved in complex chromosomal rearrangements (CCRs) and gene amplification. In this detailed molecular cytogenetic analysis, spectral karyotyping (SKY), multicolor banding (mBAND) analysis, and microarray comparative genomic hybridization (CGH) were used to refine the analysis of chromosomes with amplifications and small intrachromosomal rearrangements such as inverted duplications and interstitial deletions present in the osteosarcoma cell line MG-63. SKY analysis has limited resolving power to delineate cryptic chromosomal rearrangements, so mBAND assays were performed for a subset of chromosomes (i.e., 6, 8, 17, and 20). Of the 10 clonal CCRs analyzed in detail with mBAND, 5 were found to have rearrangements between 8q24 and either 6p23 approximately pter or 6p21, with multiple copies of this translocation inserted at various sites in the different chromosomes. In two CCRs, 6p21 and 8q24 generated an alternating pattern of mBAND probe hybridization, indicating the presence of a large coamplified repeat unit within homogeneously staining regions. Microarray CGH analysis demonstrated focal high-level amplification of 8q23 approximately q24, 6p22 approximately pter, and 6p21, in agreement with the pattern of chromosome subband gains identified with mBAND. Thus, sequential SKY, mBAND, and microarray CGH provided a comprehensive description of some of the intricate chromosomal aberrations present in the complex MG-63 karyotype and permitted reconstruction of the fine structure of the genomic rearrangements, thus providing some important mechanistic clues concerning the details of the amplification process in tumors. PMID- 15350307 TI - Genotyping of AR and PSA polymorphisms in a patient with Klinefelter syndrome, non-Hodgkin lymphoma, and adenocarcinoma of the prostate. AB - It has been hypothesized that the AR (androgen receptor) gene binds the two PSA (prostate-specific antigen) alleles with differing affinities and may differentially influence prostate cancer risk. In this article, we report a case of adenocarcinoma of the prostate in a 56-year-old man with Klinefelter syndrome (47,XXY) and non-Hodgkin lymphoma, as well as the AR and PSA genotype. AR and PSA gene polymorphisms were analyzed by polymerase chain reaction-based methods using DNA from peripheral white blood cells and the prostate cancer. We determined the methylation status of the AR gene on the X chromosome. The patient presents with the AG genotype for the ARE-1 (androgen response element) region of the PSA gene. We detect the presence of two short AR alleles with 19 and 11 CAG repeats each. Unmethylated alleles were demonstrated for both. The shorter allele was inactive in more than 60% of total DNA in both control blood and prostate cancer cells. The presence of short AR alleles and the G allele of the PSA gene may contribute to the development of prostate cancer in a 47,XXY patient. PMID- 15350308 TI - Prevalence, breakpoint distribution, and clinical correlates of t(5;12). AB - Among 56,709 cytogenetic studies performed during a 15-year period at the Mayo Clinic, 25 cases of t(5;12) were identified. Among 11 patients with available clinical information, 4 had myelodysplastic syndrome, 2 had acute myelocytic leukemia, 2 had myelofibrosis with myeloid metaplasia (MMM), 2 had atypical chronic myelocytic disorder (ACMD), and 1 had chronic myelomonocytic leukemia (CMMoL). The 5q arm was involved in all patients and the 12p arm in only two patients [ACMD,t(5;12)(q33;p13) and MMM,t(5;12)(q11.2;p11.2)], both of whom had eosinophilia and monocytosis. These two features were present in only two other patients [CMMoL,t(5;12)(q35;q24.1) and ACMD,t(5;12)(q31;q24.1)]. The t(5;12) is a rare, myelocytic-exclusive cytogenetic abnormality with a breakpoint-specific association with eosinophilia or monocytosis. PMID- 15350309 TI - Lack of DNA copy number alterations revealed with comparative genomic hybridization in fibroadenomas of the breast. AB - There are very few studies reporting DNA copy number changes in fibroadenomas of the breast. Using comparative genomic hybridization, we analyzed 20 paraffin embedded samples of fibroadenomas of the breast from patients with no familial or previous history of breast cancer. No alterations in the DNA copy number were observed in any of the tumors analyzed, regardless of the chromosomal alterations observed using conventional cytogenetic analysis. We discuss our results and compare them to other reports on fibroadenomas. PMID- 15350310 TI - The effects of BRCA1 missense variants V1804D and M1628T on transcriptional activity. AB - Many families with multiple cases of ovarian cancer, breast cancer, or both segregate inherited mutations in one allele of the tumor suppressor gene BRCA1. Genetic testing is used to assess cancer risk; however, testing can detect missense DNA alterations, called unclassified variants, of unknown functional and biological significance with uncertain risk implications. Some missense variants at the transcriptional activation domain of BRCA1 of cancer patients inactivate transcriptional activity of BRCA1, providing evidence that they are deleterious. We identified the variants V1804D and M1628T at the transcriptional activation domain of BRCA1 of two ovarian cancer patients without a family history of ovarian or breast cancer. To test if these residues are critical for transcriptional activation, we created V1804D and M1628T independently in BRCA1 cDNA via site-directed mutagenesis in a mammalian expression vector, pcDNA3.1. Wild-type, mutant, and empty vector constructs were tested in human kidney 293 cells using a p53-responsive luciferase reporter. M1628T had the same transcriptional activity as wild-type BRCA1 but V1804D and the empty vector control showed a 60% reduction. This indicates that V1804D is deleterious but M1628T is not. PMID- 15350311 TI - Acute myelocytic leukemia (M4) with t(2;4)(q22;q31) and review of the literature. PMID- 15350312 TI - Isolated interstitial 9q deletion in a case of unclassifiable myelodysplastic syndrome. PMID- 15350313 TI - Prithee, why so pale? PMID- 15350314 TI - Predictive factors for glaucomatous visual field progression in the Advanced Glaucoma Intervention Study. AB - PURPOSE: To investigate the risk factors associated with visual field (VF) progression in the Advanced Glaucoma Intervention Study (AGIS) with pointwise linear regression (PLR) analysis of serial VFs. DESIGN: Prospective, multicenter, randomized clinical trial. PARTICIPANTS: Five hundred nine eyes of 401 patients from the AGIS with a baseline VF score of or=7 VF examinations, and >or=3 years of follow-up were selected. MAIN OUTCOME MEASURE: Visual field progression. METHODS: This is a cohort study of patients enrolled in a prospective randomized clinical trial (AGIS). Worsening of a test location on PLR analysis was defined as a change of threshold sensitivity of >or=1.00 decibels a year, with P0.05). CONCLUSIONS: The prevalence of POAG in this population was 3.9%. In 92% patients with POAG, the IOP was 21 mmHg or less. PMID- 15350317 TI - Hypothyroidism and the development of open-angle glaucoma in a male population. AB - PURPOSE: To determine if hypothyroidism is associated with an increased risk of glaucoma using a large cohort of patients. DESIGN: Nested case-control study. PARTICIPANTS: Patients seen at the Veterans Affairs Medical Center in Birmingham, Alabama with newly diagnosed glaucoma between 1997 and 2001 were selected (n = 590) and age-matched to nonglaucoma controls (n = 5897). METHODS: Patient information was extracted from the Birmingham Veterans Affairs Medical Center data files containing demographic, clinical, and medication information. An index date was assigned to the glaucoma subjects corresponding to the time of diagnosis. Patients who had a glaucoma diagnosis before the observation period of the study were excluded. Ten controls were randomly selected for each patient and matched on age (+/-1 year) and an encounter on or before the index date of the matched case. MAIN OUTCOME MEASURES: Odds ratios (ORs) for the association between the prior diagnosis of hypothyroidism and the risk of developing glaucoma with adjustment for the presence of diabetes, lipid metabolism disorders, hypertension, cardiovascular disease, cerebrovascular disease, arterial disease, and migraines. RESULTS: After adjustment for the other potential risk factors, patients were significantly more likely to have prior hypothyroidism than controls (OR, 1.40; 95% confidence interval, 1.01-1.97). CONCLUSIONS: Our study has demonstrated a significantly greater risk of subjects with a preexisting diagnosis of hypothyroidism developing glaucoma, compared with controls, in a large Veterans Affairs Medical Center population. PMID- 15350318 TI - Factors associated with optic disc hemorrhages in glaucoma. AB - PURPOSE: To evaluate factors associated with optic disc hemorrhages (ODHs) in patients with open-angle glaucoma. DESIGN: Cohort study. PARTICIPANTS: One hundred thirty-seven patients with open-angle glaucoma, with an initial mean age of 60.0 years (standard deviation [SD], 11.0 years) followed up for a mean of 9.5 years (SD, 5.0 years). METHODS: The association between ODHs and various patient related variables (diabetes, systemic hypertension, heart disease, hypercholesterolemia, migraine, hypothyroidism, use of platelet antiaggregant agents) and eye-related variables (mean and range of intraocular pressure, refractive error, and severity of disease) was investigated using multivariate time-to-event analyses in patients with open-angle glaucoma. To determine the influence of the level of intraocular pressure (IOP) on the occurrence of ODHs, we compared the IOP at the time the first ODH was noted with the mean IOP of the previous 3 visits. As a control, a similar analysis was performed on the same eyes using a randomly selected visit before the occurrence of the first ODH. MAIN OUTCOME MEASUREMENTS: Patient-related and eye-related variables associated with ODHs and the comparative level of IOP in which these ODHs were observed. RESULTS: Fifty eyes of 38 patients (28%) had 1 or more ODHs during the follow-up period. The ODHs were associated with presence of diabetes (hazard ratio, 4.43; 95% confidence interval [CI], 1.8-10.50; P = 0.001) and use of aspirin (hazard ratio, 2.30; 95% CI, 1.2-4.6; P = 0.019). The IOP at the time of the first ODH was, on average, 1.4 mmHg lower than the mean IOP of the 3 previous visits (95% CI, -2.2 to -0.6 mmHg; P<0.001), whereas in the control analysis, the respective value was 0.3 mmHg lower (95% CI, -1.0 to 0.5; P = 0.410). CONCLUSIONS: Optic disc hemorrhages were associated with diabetes and aspirin use and were observed at relatively lower IOP during follow-up. PMID- 15350319 TI - Mechanism of ocular hypotensive action of bimatoprost (Lumigan) in patients with ocular hypertension or glaucoma. AB - PURPOSE: To determine the mechanism of ocular hypotensive action of bimatoprost in patients with ocular hypertension or glaucoma. DESIGN: Double-masked, placebo controlled, randomized, paired comparison crossover study of the effect of bimatoprost on aqueous humor dynamics. PARTICIPANTS AND CONTROLS: Twenty-nine patients with ocular hypertension or glaucoma. METHODS: Bimatoprost and a placebo were administered once a day, in the evening, for 7 days before assessment of aqueous dynamics using tonometry, Schiotz tonography, and fluorophotometry. Intraocular pressure (IOP) response to water drinking was measured. MAIN OUTCOME MEASURES: Aqueous humor flow rate, outflow facility, and IOP. RESULTS: Intraocular pressure was lowered 29% in the morning and 33% at noon by bimatoprost. Aqueous humor flow was unchanged. Tonographic facility of outflow was increased 47% by bimatoprost relative to the placebo. Assuming an extraocular pressure of 8 mmHg and that extraocular pressure is not altered by bimatoprost, the calculated rate of pressure-insensitive outflow was increased 95% by bimatoprost. During the first hour after water drinking, bimatoprost dampened the IOP rise. CONCLUSION: As was seen in healthy normal eyes, bimatoprost increased both the pressure-sensitive and the pressure-insensitive outflows of aqueous humor in patients with ocular hypertension or glaucoma. Bimatoprost had no significant effect on aqueous humor formation. PMID- 15350320 TI - Optic nerve head circulation in nonarteritic anterior ischemic optic neuropathy and optic neuritis. AB - PURPOSE: To quantify optic nerve head circulatory abnormalities in patients with unilateral nonarteritic anterior ischemic optic neuropathy (NAION) or optic neuritis (ON), and to assess the potential of such measurements to differentiate NAION from ON. DESIGN: Prospective, cross-sectional, observational study. PARTICIPANTS: Thirty consecutive patients with unilateral NAION, 22 consecutive patients with unilateral ON, and 50 healthy control subjects. METHODS: All subjects underwent a complete neuro-ophthalmologic evaluation. The widths of Doppler-broadened frequency spectra, which are directly proportional to the speed of blood cells flowing through the capillaries of the optic nerve head, were measured at multiple sites in both eyes of each subject. The variation of Doppler broadening (DB) with age was determined in the control subjects. Doppler broadening values in the patients were compared between similar sites in affected and contralateral eyes, and between both affected and contralateral eyes and the age-adjusted values determined in the control subjects. MAIN OUTCOME MEASURES: The differences in DB between (1) the affected and contralateral eyes of the patients, (2) the patients and the control subjects, and (3) the patients with NAION and those with ON. RESULTS: In NAION, DB was decreased at both temporal ( 20.2% and -18.5%) and nasal (-12.8% and -12.4%) sites of the nerve head in affected eyes compared with contralateral eyes or eyes of control subjects. In ON, DB was also decreased at temporal sites (-11.3% and -9.2%) in affected eyes compared with contralateral or control eyes. At nasal sites, there were no significant differences in DB in affected eyes of ON patients compared with contralateral or control eyes. The DB decreases were significantly greater in NAION patients than in ON patients. CONCLUSIONS: Optic nerve head circulatory abnormalities are present in patients with NAION or ON. This is the first demonstration of such abnormalities in ON, a finding consistent with the recent attention given to the phenomenon of axonal loss in this disease. Although there are differences in the circulatory abnormalities between the 2 diseases that provide insights into the pathophysiological mechanisms at play, they are not large enough to enable the clinician to distinguish between ON and NAION in an individual patient. PMID- 15350321 TI - Swedish interactive thresholding algorithm fast for following visual fields in prepubertal idiopathic intracranial hypertension. AB - OBJECTIVE: To examine the reliability of Swedish Interactive Thresholding Algorithm Fast (SITA Fast) visual fields (VFs) in prepubertal idiopathic intracranial hypertension (IIH) and to compare whether age, gender, or severity of visual outcome influenced the reliability of VF tests. DESIGN: Prospective, longitudinal cohort study. PARTICIPANTS: Twenty-six prepubertal children (<11 years of age; mean age, 7.2 years) with IIH. TESTING: Children were prospectively followed up using a child-oriented program, which included SITA Fast VF tests. Age, gender, and severity of visual outcome were correlated with reliability of performance on SITA Fast VFs using a 1-way analysis of variance, point-biserial correlation, and the chi-square test for independence of observation. MAIN OUTCOME MEASURES: Statistical analyses results that correlated the reliability of SITA Fast VFs with age, gender, and visual outcome. RESULTS: Three children were treated at the age of 2 years and were unable to perform automated VF tests. Four children were treated at the age of 4 years, 3 of whom were able to perform repeatable reliable SITA Fast VF tests. Forty-two eyes of 21 children had an average SITA Fast test time of 4.5 minutes for each eye, compared with 8 minutes using the Fastpac strategy. Age was not associated with reliability scores of SITA Fast tests (F = 0.971, not significant [ns]). Gender did not influence the reliability of SITA Fast VF tests (chi-square((1)) = 0.669, ns), nor did severity of visual outcome (chi-square((2)) = 3.348, ns). Visual deficits were observed in 55% of patients at presentation and in 27% of patients after resolution of papilledema. CONCLUSIONS: The SITA Fast VF tests can be performed from age 4 years and offer a reliable method for shortening test time. A child-oriented follow-up program, which entails a shortened testing time, may improve outcome in prepubertal IIH. PMID- 15350322 TI - Comparison of deep lamellar keratoplasty and penetrating keratoplasty in patients with keratoconus. AB - OBJECTIVE: To compare the therapeutic outcomes after deep lamellar keratoplasty (DLK) and penetrating keratoplasty (PK) in patients with keratoconus. DESIGN: Retrospective case-control study. PARTICIPANTS: We reviewed the clinical notes of 47 patients diagnosed clinically with keratoconus who had received DLK (26 eyes of 25 patients) or PK (25 eyes of 22 patients) at Moorfields Eye Hospital or the Royal Shrewsbury Hospital between 1994 and 2001. The patients in the 2 groups were matched for severity of their keratoconus by preoperative visual acuity. METHODS: Deep lamellar keratoplasty was performed with the Melles technique in 7 eyes and the technique described by Sugita and Kondo in 19 eyes. Penetrating keratoplasty was performed with a standard technique using a Hessburg-Barron trephine. A single continuous 16-bite 10-0 nylon suture was placed and adjusted in both groups. MAIN OUTCOME MEASURES: Best-corrected visual acuity (BCVA), refractive results, surgical techniques for DLK, and complication rates were analyzed. RESULTS: The 25 patients with keratoconus who underwent DLK had a mean age of 32.6 years and a median follow-up of 28 months. The mean age of the 22 patients who underwent PK for keratoconus was 34 years. This group was followed up for a median time of 55 months. The median final BCVA of patients in the DLK group was 6/9 and in the PK group 6/6 (no statistical significance). The median result for the final spherical equivalent power in both groups was mild myopia, although the DLK group had more myopia, and the median astigmatism was less than 5.00 diopters cylinder for both groups. Complication rates were similar for DLK and PK, although the nature of the complications varied. CONCLUSIONS: Penetrating keratoplasty is no longer an automatic choice for the surgical treatment for keratoconus; DLK seems to be a safe alternative. Best-corrected visual acuity, refractive results, and complication rates are similar after DLK and PK. Deep lamellar keratoplasty is more technically challenging but allows the risk of endothelial rejection to be avoided and may reduce the risk of late endothelial failure. PMID- 15350323 TI - United States Food and Drug Administration clinical trial of the Implantable Collamer Lens (ICL) for moderate to high myopia: three-year follow-up. AB - PURPOSE: To report on 3-year postoperative safety and efficacy outcomes with the Myopic Implantable Collamer Lens (ICL). DESIGN: Prospective nonrandomized clinical trial. PARTICIPANTS: Five hundred twenty-six eyes of 294 patients with between 3.0 and 20.0 diopters (D) of myopia participating in the United States Food and Drug Administration clinical trial of the ICL for myopia. INTERVENTION: Implantation of the ICL. MAIN OUTCOME MEASURES: Uncorrected visual acuity (VA), refraction, best spectacle-corrected VA (BSCVA), adverse events, operative and postoperative complications, lens opacity analysis, subjective satisfaction, and patient symptoms. RESULTS: At 3 years, 59.3% had 20/20 or better VA, and 94.7% had 20/40 or better uncorrected VA if BSCVA was 20/20 and patients were targeted for emmetropia; 67.5% of patients were within 0.5 D and 88.2% were within 1.0 D of predicted refraction. The mean improvement in BSCVA ranged between 0.5 and 0.6 lines. At 3 years postoperatively, 3 eyes (0.8%) decreased by >or=2 lines of BSCVA, in contrast to 40 eyes (10.8%) that improved by a similar amount. Contrast sensitivity improved postoperatively. Cumulative 3-year corneal endothelial cell loss was under 10%. Early largely asymptomatic, presumably surgically induced anterior subcapsular opacities (trace or greater) were seen in 14 eyes (2.7%), with only 2 being clinically significant. Five eyes (0.9%) of 3 patients developed nuclear opacities of grade >2 at 2 to 3 years postoperatively. Three (0.6%) ICL removals with cataract extraction and IOL implantation have been performed. Only 0.6% reported dissatisfaction; 97.1% of patients reported they would choose ICL implantation again. Incidences of patient symptoms, glare, halos, double vision, night vision problems, and night driving difficulties decreased or remained unchanged after ICL surgery. CONCLUSION: Three-year results from this standardized, multicenter clinical investigation support the safety, efficacy, and predictability of ICL surgery to treat moderate to high myopic refractive errors. PMID- 15350324 TI - Comparison of various pupil dilatation methods for phacoemulsification in eyes with a small pupil secondary to pseudoexfoliation. AB - PURPOSE: To compare 4 methods for intraoperative pupil dilatation in eyes with pseudoexfoliation syndrome and insufficient pupil size during phacoemulsification. DESIGN: Prospective, randomized, comparative, interventional case series. PARTICIPANTS: Forty eyes of 40 patients with pseudoexfoliation and maximally dilated pupil size smaller than 3.5 mm. INTERVENTION: Mechanical pupil dilatation with iris-retractor hooks (group I), polymethyl methacrylate (PMMA) pupil dilator-ring (Morcher, Stuttgart, Germany) (group II), Beehler pupil dilator (group III), and bimanual stretching (group IV). MAIN OUTCOME MEASURES: Performance (pupil size achieved), complications, and added surgical time. RESULTS: The mean pupil sizes achieved with the PMMA pupil-dilator ring, Beehler pupil dilator, and bimanual stretching were 5.9+/-0.6 mm, 5.5+/-0.8 mm, and 4.9+/ 0.7 mm, respectively. A square-shaped pupil was achieved with iris-retractor hooks, and the mean size of the largest circle that can fit in this square was 5.6+/-0.6 mm. There were no statistically significant differences in the postdilatation pupil sizes between the 4 study groups (P>0.05). Apart from self limited intraoperative hemorrhage from pupil margin, iris sphincter rupture was the only observed complication related to mechanical pupil dilatation. This occurred in 4 eyes in groups I and III, 3 eyes in group IV, and 1 eye in group II (P>0.05). The mean added surgical time for placement of iris-retractor hooks and for implantation of a PMMA pupil-dilator ring was 297+/-51 and 176+/-54 seconds, respectively. The additional time required for pupil dilatation with the Beehler pupil dilator and bimanual stretching was 65+/-8 and 55+/-10 seconds, respectively. The time needed for pupil dilatation in groups I and II is significantly longer than that in groups III and IV (P<0.001). CONCLUSIONS: All 4 methods used in this study were effective procedures for the mechanical dilatation of small pupils secondary to pseudoexfoliation syndrome. Iris retractor hooks and the PMMA pupil-dilator ring are the most time-consuming techniques but have the advantage of a stable pupil size throughout the surgery. The PMMA pupil-dilator ring causes the least iris trauma. The Beehler pupil dilator and bimanual stretching technique were the least time-consuming methods for mechanical pupil dilatation. PMID- 15350325 TI - Central surface curvatures of postmortem- extracted intact human crystalline lenses: implications for understanding the mechanism of accommodation. AB - PURPOSE: To measure the radii of curvature of postmortem, whole, encapsulated human crystalline lenses, free of all zonular attachments, and to calculate their corresponding optical powers. DESIGN: Experimental study. PARTICIPANTS: Thirty human crystalline lenses from donors with a mean age of 33.6+/-14.4 years. METHODS: Intact clear human crystalline lenses were obtained within an average of 21 hours of death. The lenses were removed from the eye by the contributing eye bank and shipped in Optisol-GS, a physiologic preservative storage medium. These lenses, with intact capsules, were freed of all zonular attachments. The lenses were stored at 7 degrees C and were maintained in the same storage medium during the period that they were held for evaluation. Using a portable Keratron Scout corneal topographer (Eyequip, Ponte Vedra Beach, FL) fixed to an optical bench, the radii of curvatures of the anterior and posterior surfaces of the crystalline lens were measured daily for 10 days after receipt of the tissue. The capsules of the crystalline lenses remained intact, and the lenses were clear throughout the study. Measurements were made at room temperature after removing the lens from storage. Eight repetitions of the topography were made from each surface on each day to determine the accuracy and stability of the measurement. Profile photographs were taken daily to establish the central crystalline lens thickness. The corresponding optical power of each physiologically maintained crystalline lens was calculated. MAIN OUTCOME MEASURES: The main outcome measures were the central anterior radius of curvature, the central posterior radius of curvature, the central thickness of each crystalline lens, and the amount of change in these parameters over 10 days. RESULTS: The means +/- standard deviations of the central anterior and posterior radii of curvatures of the 30 adult lenses were 10.5+/-0.6 mm and 7.1+/-1.0 mm, respectively. The mean +/- standard deviation of the central thickness, as measured from profile photographs, was 3.9+/-0.5 mm. These mean measurements remained stable for up to 5 days after receipt of the tissue. The mean and standard deviation of the calculated optical power of these postmortem, physiologically maintained, intact human crystalline lenses from donors with a mean age of 33.6 years was 19.8+/-1.7 diopters. CONCLUSIONS: Lenses free of zonular tension are at an optical power that would be associated with optimal distance acuity. PMID- 15350326 TI - Time course of development of posterior vitreous detachment in the fellow eye after development in the first eye. AB - PURPOSE: To determine when posterior vitreous detachment (PVD) develops in the fellow eye after development in the first eye. DESIGN: Prospective, observational case series. PARTICIPANTS: Fifty-one patients with acute unilateral PVD participated in the study. METHODS: All patients underwent a comprehensive ocular examination, including a dynamic vitreous examination using biomicroscopy with and without a preset lens and retinal examination with scleral depression, every 3 months at least until PVD developed in the fellow eye. RESULTS: Posterior vitreous detachment developed in 4 eyes (8%) within 6 months, 12 eyes (24%) within 1 year, 24 eyes (47%) within 18 months, 33 eyes (65%) within 2 years, 41 eyes (80%) within 30 months, and 46 eyes (90%) within 3 years. In 5 eyes (10%), PVD did not develop during the follow-up period of more than 3 years. CONCLUSIONS: Posterior vitreous detachment develops frequently in the fellow eye from 6 months to 2 years after development in the first. These findings are useful for the management of patients with unilateral PVD. PMID- 15350327 TI - Visual outcome after discontinuation of corticosteroids in atypical severe central serous chorioretinopathy. AB - PURPOSE: To elucidate the effect of discontinuation of corticosteroids in patients with atypical severe central serous chorioretinopathy (CSC) on retinal reattachment, resolution of retinal pigment epithelium (RPE) leaks, and improvement in visual acuity (VA). DESIGN: Prospective, noncomparative, observational case series. PARTICIPANTS: Twenty-four eyes of 17 patients who were being treated with corticosteroids for atypical severe CSC. Of these 17 patients, 16 were treated inappropriately with corticosteroids for their ocular condition; presumably, these patients' conditions were misdiagnosed, and they were thought to have choroiditis, Harada's syndrome, or similar entities, and not central serous chorioretinopathy. INTERVENTION: Observation or laser photocoagulation. MAIN OUTCOME MEASURES: Reattachment of the retina, obliteration of RPE leaks on fundus fluorescein angiography, and improvement in Snellen VA. RESULTS: Discontinuation of corticosteroids resulted in reattachment of the retina in 21 eyes (87.5%), with median time to reattachment of 49 days (range, 32-400); only 3 eyes required laser photocoagulation. Fundus fluorescein angiography showed obliteration of RPE leaks at a median period of 75 days (range, 32-400) in the observed eyes; the median VA improved from 20/80 to 20/30. The mean follow-up was 16.5 months. CONCLUSIONS: Discontinuation of corticosteroids in atypical CSC helped in obliteration of RPE leaks and retinal reattachment in 87.5% of the eyes without laser treatment, and improvement in VA was observed. PMID- 15350328 TI - Change in visual function after macular translocation with 360 degrees retinectomy for neovascular age-related macular degeneration. AB - OBJECTIVE: To measure the change in vision and visual outcomes at 12 months after macular translocation with 360 degrees retinectomy (MT360) and silicone oil tamponade in patients with bilateral vision loss resulting from subfoveal choroidal neovascular membranes in age-related macular degeneration (AMD). DESIGN: A prospective, interventional, consecutive, noncomparative case series. PARTICIPANTS: Sixty-four patients with bilateral vision loss resulting from neovascular AMD. METHODS: Eligible patients had AMD with subfoveal choroidal neovascularization in the operative eye and a maximum of 6 months of central vision loss. Preoperative and 12-month postoperative evaluations included standardized testing of near and distance acuity and reading speed. Patients underwent MT360 with silicone oil tamponade, followed 2 months later by extraocular muscle surgery and silicone oil removal. MAIN OUTCOME MEASURES: Change in distance acuity, near acuity, and reading speed at 12 months after MT360 compared with those values before surgery. RESULTS: Sixty-one patients were followed up for 12 months. All eyes were translocated successfully. Median distance acuity letter score improved from 62 letters (Snellen equivalent of approximately 20/125) before surgery to 69 letters (approximately 20/80) by 12 months after surgery (P = 0.03). Median near acuity improved from 0.70 logarithm of the minimum angle of resolution (logMAR) units (approximately 20/100) before surgery to 0.44 logMAR units (approximately 20/55) at 12 months (P<0.001). Median reading speed improved from 71 words per minute (wpm) before surgery to 105 wpm at 12 months after surgery (P<0.001). At 12 months, distance acuity improved by 1 or more lines in 32 patients (52%). In patients with either preoperative distance or near acuity of 20/80 or better, 74% and 95% of patients, respectively, remained in this range of acuity. In patients with either preoperative distance or near acuity of worse than 20/80, 40% and 48% of patients, respectively, improved to 20/80 or better. Postoperative retinal detachment developed in 5 patients (8%), with the macula involved in 2 patients, and all retinas were reattached successfully. CONCLUSIONS: Macular translocation with 360 degrees retinectomy with silicone oil tamponade is effective in significantly improving visual function in patients with neovascular AMD, as demonstrated by the improvement in distance and near acuity and reading speed at 12 months after surgery in these patients. Although this is a complex surgical intervention, patients with preoperative visual acuity of 20/80 or better at near or distance are highly likely to retain the 20/80 or better acuity at 12 months after surgery. Macular translocation with 360 degrees retinectomy is an effective treatment option for patients with vision loss in their second eye resulting from neovascular AMD. PMID- 15350329 TI - Photodynamic therapy with verteporfin in ocular histoplasmosis: uncontrolled, open-label 2-year study. AB - OBJECTIVE: To evaluate the safety, effect on visual function, and fluorescein angiographic appearance of subfoveal choroidal neovascularization (CNV) through 2 years after photodynamic therapy with verteporfin (Visudyne; Novartis AG, Basel, Switzerland) in patients with ocular histoplasmosis syndrome (OHS). DESIGN: Open label, 3-center, uncontrolled clinical study. PARTICIPANTS: Ocular histoplasmosis syndrome patients with subfoveal CNV (N = 26) with a greatest linear dimension no larger than 5400 microm with classic or occult CNV extending under the geometric center of the fovea, and best-corrected visual acuity letter score of approximately 20/40 to 20/200. METHODS: The methods were similar to those described in the 1-year results with follow-up examinations every 3 months continuing through the second year. During the second year, additional therapy was recommended if fluorescein angiography showed leakage at a scheduled visit. MAIN OUTCOME MEASUREMENTS: Visual function measurements included the changes from baseline in visual acuity and contrast sensitivity scores. Lesion size and leakage from classic and occult CNV were assessed at month 12 and month 24. Safety assessments also were made. RESULTS: A 24-month examination was completed in 22 of the 26 enrolled participants (85%). At the 24-month examination, median improvement from baseline in visual acuity of the 22 patients evaluated was 6 letters; median contrast sensitivity improved by 3.5 letters. At the 24-month examination, 10 patients (45%) gained 7 or more letters of visual acuity from baseline, whereas 4 patients (18%) lost 8 or more letters, including 2 patients (9%) who lost at least 15 letters. There was absence of fluorescein angiographic leakage from classic CNV in 17 of the 20 evaluable lesions (85%), and leakage from occult CNV was absent in all eyes. No serious ocular adverse events were reported, and no serious systemic event was considered to be associated with treatment. CONCLUSIONS: Median visual acuity improved and fluorescein angiographic leakage decreased after verteporfin therapy in this small, uncontrolled clinical study of patients with subfoveal CNV resulting from OHS. Verteporfin therapy seemed to be relatively safe in these patients. The selected cases feature fluorescein angiographic examples of CNV that are important in determining when to apply verteporfin therapy. PMID- 15350330 TI - Intraocular pressure variations after diode laser photocoagulation for threshold retinopathy of prematurity. AB - PURPOSE: To investigate the intraocular pressure (IOP) variations occurring after indirect diode laser photocoagulation for threshold retinopathy of prematurity. DESIGN: Prospective, nonrandomized, comparative study. PARTICIPANTS: A study group-21 consecutive premature babies (42 eyes) undergoing diode laser photocoagulation for retinopathy of prematurity-and control groups-32 premature babies (64 eyes) undergoing retinal examination with or without scleral indentation. INTERVENTION: Intraocular pressure was measured with a portable electronic tonometer before, immediately after, and 1, 2, and 3 days after diode laser photocoagulation in the study group; before retinal examination in control group 1; and before and after retinal examination with scleral indentation in control group 2. MAIN OUTCOME MEASURE: Intraocular pressure after diode laser photocoagulation for threshold retinopathy of prematurity. RESULTS: Mean IOP in the study group rose from 15 mmHg (standard deviation [SD] = 4.1) before coagulation to 33.2 mmHg (SD = 7.8; range, 20-50) immediately after, and then dropped to 22.2 (SD = 5.2), 16.5 (SD = 3), and 14.5 mmHg (SD = 2.1) 1, 2, and 3 days later, respectively. All the changes were statistically significant at P<0.0001, except for the difference between days 2 and 3 (P = 0.096). Mean baseline IOPs were 16.3 mmHg (SD = 3.7) in control group 1 and 15.7 mmHg (SD = 2.3) in control group 2 (P = 0.84 between control group 1 and study group, and P = 0.32 between control group 2 and the study group). At termination of the retinal examination with scleral indentation (control group 2), IOP measured 15.1 mmHg (SD = 2.2) (P = 0.49 compared with baseline). CONCLUSIONS: Intraocular pressure may be significantly elevated after diode laser photocoagulation for retinopathy of prematurity. The mechanism and long-term clinical implications of this observation should be investigated. PMID- 15350331 TI - Melanocytoma of optic disc in 115 cases: the 2004 Samuel Johnson Memorial Lecture, part 1. AB - PURPOSE: To report clinical variations and the natural course of melanocytoma of the optic disc. PATIENTS AND METHODS: A retrospective study was conducted on a consecutive series of patients with melanocytoma of the optic disc who were evaluated at the Ocular Oncology Service at Wills Eye Hospital. Demographic information and variations in appearance and size of the tumor and local ocular complications were noted. Frequency of tumor growth and malignant transformation was recorded. Clinical factors, including patient data and tumor features, were analyzed for their impact on visual acuity loss of 2 or more Snellen lines and the tumor growth, using Cox proportional hazards regression models. Kaplan-Meier survival estimates of probability of visual acuity loss of 2 or more Snellen lines and probability of tumor growth were performed as a function of time from the initial examination. MAIN OUTCOME MEASURES: Visual acuity loss of 2 or more Snellen lines, tumor growth, and malignant transformation of the tumor. RESULTS: There were 115 patients (116 eyes) with melanocytoma of the optic disc. The mean age at diagnosis was 50 years; 38% of patients were male and 62% of patients were female; 65% of patients were white, 29% of patients were African American, and 6% of patients were Asian, Hispanic, Indian, or Arabic. The lesion was unilateral in 99% of patients. Visual symptoms that seemed to be related to the melanocytoma were present in 24% of patients, and an afferent pupillary defect was noted in 9% of patients. Associated ocular abnormalities included ocular melanocytosis (8%), racial melanosis (7%), optic nerve hypoplasia (2%), and retinitis pigmentosa (1%). The melanocytoma was dark brown to black in 100% of patients. The mean tumor diameter was 2 mm and the mean thickness was 1 mm. Associated findings included a choroidal component (54%), retinal component (30%), optic disc edema (25%), retinal edema (16%), localized subretinal fluid (14%), retinal exudation (12%), retinal hemorrhage (5%), vitreous seeds (4%), and retinal vein obstruction (3%). Fluorescein and indocyanine green angiography typically showed persistent hypofluorescence of the lesion. Using Kaplan-Meier survival curves, related visual loss occurred in 18% of patients by 10 years, and minor tumor enlargement occurred in 11% of patients by 5 years and in 32% of patients by 10 years. Malignant transformation was documented in 2 patients (2%). CONCLUSIONS: Although melanocytoma of the optic disc generally is considered to be a benign, stationary lesion, it can produce several local complications, can cause visual loss, can grow slowly, and, rarely, can undergo malignant transformation into melanoma. Patients with optic disc melanocytoma should undergo periodic ocular examination. PMID- 15350332 TI - Clinical survey of 1643 melanocytic and nonmelanocytic conjunctival tumors. AB - OBJECTIVE: To report the spectrum and frequency of melanocytic and nonmelanocytic conjunctival tumors in an ocular oncology practice. DESIGN: Retrospective noninterventional case series. PARTICIPANTS: One thousand six hundred forty-three consecutive patients with a conjunctival mass evaluated at an ocular oncology department. METHODS: A chart review was conducted to obtain the clinical features of the patient and tumor and to tabulate and categorize the diagnoses. MAIN OUTCOME MEASURES: Tumor diagnosis overall and relative to patient age, race, and gender and relative to tumor location and laterality. RESULTS: In 1643 consecutive patients, the tumor was classified as melanocytic in 872 cases (53%) and nonmelanocytic in 771 cases (47%). The nonmelanocytic categories included congenital choristomatous (n = 40 [2%]), epithelial (n = 219 [13%]), vascular (n = 63 [4%]), fibrous (n = 7 [<1%]), neural (n = 1 [<1%]), xanthomatous (n = 1 [<1%]), myxomatous (n = 1 [<1%]), lipomatous (n = 23 [1%]), lacrimal gland origin (n = 12 [<1%]), lymphoid (n = 128 [8%]), leukemic (n = 3 [<1%]), metastatic (n = 13 [<1%]), secondary (n = 54 [3%]) tumors, and non-neoplastic lesions simulating a tumor (n = 206 [13%]). Of the 872 melanocytic lesions, the specific tumor diagnosis was nevus in 454 cases (52%), melanoma in 215 (25%), and primary acquired melanosis in 180 (21%). Patients with choristomatous, vascular, fibrous, xanthomatous, and myxomatous tumors presented at a mean age of <40 years, and those with malignant epithelial, lipomatous, leukemic, and secondary tumors presented at a mean age of >60. Of the 219 patients with epithelial tumors, 80% occurred in males, whereas the incidence of melanocytic lesions was equal in males and females. African-American patients represented only 7% of epithelial tumors, <1% of melanomas, and 8% of lymphoid tumors. CONCLUSION: Conjunctival tumors were of melanocytic origin in 53% of cases and nonmelanocytic origin in 47%. Overall, melanocytic tumors, epithelial tumors, and lymphoid tumors accounted for 74% of all cases. These tumors were far more common in Caucasian patients, and epithelial tumors were found more frequently in men. PMID- 15350333 TI - Treatment of recurrent corneal and conjunctival intraepithelial neoplasia with topical interferon alfa 2b. AB - OBJECTIVE: To evaluate topical interferon alfa 2b (IFNalpha2b) as a single therapeutic agent in the treatment of presumed recurrent corneal and conjunctival intraepithelial neoplasia. DESIGN: Noncomparative, retrospective case series. PARTICIPANTS: Seven consecutive patients with recurrent corneal and conjunctival intraepithelial neoplasia diagnosed at the University of Minnesota from July 2000 to November 2003 were studied retrospectively. All patients had a history of histologically proven primary corneal and conjunctival intraepithelial neoplasia and were treated by surgery, cryotherapy, radiation, and/or topical mitomycin C before recurrence. INTERVENTION: Patients with a clinical diagnosis of recurrent corneal and conjunctival intraepithelial neoplasia were treated with recombinant topical IFNalpha2b drops (1 million IU/ml) 4 times daily until lesion resolution was noted. MAIN OUTCOME MEASURES: A review of medical records was performed to assess the duration of and response to treatment with topical IFNalpha2b, defined by clinical resolution of corneal and conjunctival intraepithelial neoplasia. RESULTS: The average age of the 7 patients at the initiation of topical IFNalpha2b treatment for presumed recurrent corneal and conjunctival intraepithelial neoplasia was 68.7 years (range, 54-88). Six of 7 patients had successful treatment of recurrent corneal and conjunctival intraepithelial neoplasia lesions with topical IFNalpha2b treatment. The average length of IFNalpha2b treatment to resolution of recurrent corneal and conjunctival intraepithelial neoplasia was 14.5 weeks (range, 5-24). After treatment with topical IFNalpha2b for recurrent corneal and conjunctival intraepithelial neoplasia, 2 patients had another recurrence of corneal and conjunctival intraepithelial neoplasia, noted at 1 year and 2 months, respectively. The average post-treatment follow-up was 11.7 months (range, 8-17) after the resolution of recurrent corneal and conjunctival intraepithelial neoplasia. No side effects of treatment were noted in any patient. CONCLUSIONS: Topical IFNalpha2b as a single therapeutic agent is an effective treatment of presumed recurrent corneal and conjunctival intraepithelial neoplasia. It offers the benefits of topical therapy and avoids the risks of surgical or other interventions-specifically, ocular surface toxicity, cicatricial conjunctival changes, and limbal stem cell deficiency. Larger controlled studies with longer follow-up periods are recommended to confirm the long-term efficacy and safety of this topical treatment. PMID- 15350334 TI - Primary intraocular lymphoma: clinical, cytologic, and flow cytometric analysis. AB - PURPOSE: To compare cytologic with flow cytometric results of vitreous biopsy specimens obtained to rule out primary intraocular lymphoma (PIOL). STUDY DESIGN: Prospective noncomparative case series. PARTICIPANTS: Patients suspected of having PIOL who underwent vitreous biopsy were evaluated. METHODS: Patients underwent a standard 3-port vitrectomy and vitreous biopsy to rule out PIOL. Each undiluted specimen was split, and half was prepared for cytologic evaluation with the collodion bag method; the other half was submitted for flow cytometric immunophenotyping (FCI). The diluted specimen was processed as a cell block for cytology. MAIN OUTCOME MEASURES: Final diagnosis based on cytology and FCI. RESULTS: Ten of 14 patients had sufficient specimens for both cytologic and FCI evaluation. Three patients had chronic inflammation confirmed by both methods. Six patients had large cell lymphoma identified by both cytology and FCI. Two of those 6 patients initially had insufficient specimen for FCI. One patient had large cell lymphoma diagnosed cytologically that was initially negative for a clonal population by FCI. All lymphomas were B-cell type. CONCLUSIONS: Cytologic evaluation is an accurate diagnostic technique to evaluate for PIOL. FCI is useful for immunophenotyping PIOL. Multiple biopsies may be required to achieve a diagnosis. PMID- 15350335 TI - Ocular complications after iodine brachytherapy for large uveal melanomas. AB - PURPOSE: To evaluate anterior and posterior segment complications and their management after iodine 125 plaque brachytherapy (IBT) for large uveal melanoma. DESIGN: Retrospective nonrandomized interventional study. PARTICIPANTS: Ninety six patients with a large uveal melanoma according to the Collaborative Ocular Melanoma Study criteria. METHODS: The patients underwent primary IBT (median dose to tumor apex, 87 Gy). The median tumor height and diameter were 10.7 mm (range, 4.5-16.8) and 16.5 mm (range, 7.3-25.0), respectively, and the median follow-up time was 3.5 years (range, 0.3-10.4). Cumulative incidence analysis and competing risks regression were used to analyze the time to individual complications and to identify risk factors. Death and secondary enucleation were analyzed as competing risks. MAIN OUTCOME MEASURES: Cataract, iris neovascularization, glaucoma, maculopathy, optic neuropathy, vitreous hemorrhage, and persistent exudative retinal detachment (RD). RESULTS: The 5-year cumulative incidences of cataract, iris neovascularization, and glaucoma were 69% (95% confidence interval [CI], 57% 78%), 62% (95% CI, 50%-71%), and 60% (95% CI, 48%-70%), respectively. Posterior segment complications were less common. The 5-year incidences of maculopathy and optic neuropathy were 52% (95% CI, 35%-65%) and 46% (95% CI, 30%-61%), and those of vitreous hemorrhage and persistent RD were 36% (95% CI, 23%-48%) and 25% (95% CI, 15%-36%), respectively. More than 80% of complications were diagnosed within 3 years. Cataract was the earliest complication to appear. Except for cataract, the cumulative incidence of dying without developing a particular complication was 0.24 to 0.62 times that of first developing the complication. Increasing tumor height, which correlates to increasing doses to adjacent tissues, was associated with time to cataract (P = 0.017), iris neovascularization (P = 0.087), and RD (P = 0.046). Maculopathy and optic neuropathy were associated primarily with distance to the fovea (P = 0.015) and optic disc (P = 0.015), respectively. Of 57 patients with cataract, 47% underwent cataract extraction, and 12% of 51 patients with glaucoma were treated with cyclophotocoagulation. The prevalences of cataract, elevated intraocular pressure, and RD were 43%, 39%, and 13%, respectively, at 5 years. CONCLUSIONS: The frequency with which ocular complications develop after IBT is notably influenced by competing risks. Cumulative incidence and prevalence analysis provide realistic estimates for patient counseling. PMID- 15350336 TI - Ocular trauma in a rural south Indian population: the Aravind Comprehensive Eye Survey. AB - PURPOSE: To determine the rate of ocular trauma in a rural population of southern India and its impact on vision impairment and blindness. METHODS: A population based cross-sectional study of 5150 persons 40 years or older in a randomly chosen rural population of 3 districts of southern India. Prospective information on trauma, type and agent of injury, setting of injury, and details of treatment sought for the last episode was recorded with questionnaires after face-to-face interviews. All interviewed subjects underwent a comprehensive ocular examination, including vision estimations, slit-lamp biomicroscopy examinations, and dilated posterior segment examinations. RESULTS: We elicited a history of ocular trauma in either eye from 229 (4.5%) persons, including 21 (0.4%) persons with bilateral ocular trauma. Blunt injuries (n = 124; 54.9%) were the major cause for trauma reported in this population. The most common setting where the ocular trauma occurred was during agricultural labor (n = 107; 46.9%). Nearly three quarters (n = 170; 74.2%) of those reporting ocular trauma sought treatment from an eye specialist (n = 104; 57.8%) and one fifth (n = 37; 20.6%) from a traditional healer. The age-adjusted (adjusted to the population estimates for India for the year 2000) prevalence for blindness in any eye caused by trauma was 0.8% (95% confidence interval [CI], 0.4-1.1). The odds ratios (OR) for trauma were higher for males (OR, 2.2; 95% CI, 1.6-3.0) and laborers (OR, 1.7; 95% CI, 1.2-2.4) and lower for literates (OR, 0.7; 95% CI, 0.5- 0.9). Seeking treatment from a traditional eye healer for trauma was not associated with vision impairment (OR, 1.0; 95% CI, 0.3-3.2) or with blindness (OR, 3.4; 95% CI, 0.2 56.5). CONCLUSIONS: Eye care programs may need to consider ocular trauma as a priority in this population, because the lifetime prevalence of ocular trauma is higher than that reported for glaucoma, age-related macular degeneration, or diabetic retinopathy from this population. Simple measures such as education regarding the use of protective eyewear could possibly significantly decrease this preventable cause of visual disability. PMID- 15350337 TI - Lacrimal outflow dysgenesis. AB - PURPOSE: To evaluate patients with lacrimal dysgenesis. DESIGN: Retrospective, nonrandomized, comparative case series. PARTICIPANTS: Fifty patients with lacrimal dysgenesis managed between 1992 and 2003. TESTING/INTERVENTION: The diagnosis of lacrimal outflow dysgenesis was made based on the following criteria: absent or hypoplastic punctum, canaliculus, lacrimal sac, and nasolacrimal duct, appearing in isolation or combination. Our management algorithm included observation, punctoplasty with intubation, or open lacrimal surgery, with or without intubation. MAIN OUTCOME MEASURE: Success, partial success, or failure of treatment. RESULTS: Lacrimal dysgenesis in our 50 patients (23 male and 27 female) involved 83 eyes, distributed as follows: proximal in 74 eyes (89%), distal in 27 (33%), and both in 18 (22%). Thirty (60%) patients presented with isolated lacrimal dysgenesis, and 20 (40%) presented with a systemic syndrome or dysmorphism. Thirty-three (66%) patients had bilateral involvement. Epiphora was the most common presenting symptom. Eighteen (36%) patients had a positive family history. Open lacrimal surgery was performed in a total of 29 (35%) of the 83 eyes: 25 had success, 3 had partial success, and 1 had failure. CONCLUSION: Both sporadic and hereditary forms of lacrimal outflow dysgenesis may present as an isolated finding or a part of a systemic syndrome or dysmorphism, occurring usually with bilateral involvement and presenting at a younger age in the setting of systemic anomalies. Proximal and distal lacrimal outflow systems may be involved with epiphora as the most common presenting symptom. The algorithm we have presented provides a systematic approach to the management of lacrimal outflow dysgenesis. PMID- 15350338 TI - Internal limiting membrane peeling. PMID- 15350339 TI - Biostatistics. PMID- 15350341 TI - Ultrastructure of the internal limiting membrane. PMID- 15350343 TI - Suspected child abuse victims. PMID- 15350345 TI - Suspected child abuse victims. PMID- 15350346 TI - Balloon dacryoplasty. PMID- 15350348 TI - Radical trachelectomy: is it here to stay? PMID- 15350349 TI - Vaginal radical trachelectomy: an oncologically safe fertility-preserving surgery. An updated series of 72 cases and review of the literature. AB - OBJECTIVE: To review the oncological results and complication rate of our first consecutive 72 completed cases of vaginal radical trachelectomies (VRT). METHODS: From October 1991 to October 2003, we have planned 82 VRT in patients with early stage cervical cancer (stages IA, IB, and IIA). The VRT was preceded by a complete laparoscopic pelvic node dissection and laparoscopic parametrectomy. RESULTS: The planned procedure was successfully completed in 72 cases and was abandoned in 10 cases (12%) because of either positive nodes discovered at the time of surgery (4), positive endocervical margins (5) or extensive tubal adhesions (1). The median age of the remaining 72 patients was 31 and most (75%) were nulliparous. The majority of the lesions were stage IA2 (32%) or IB1 (60%) and 54% were grade 1. In terms of histology, 58% were squamous and 42% were adenocarcinomas. Vascular space invasion was present in 20% of cases, and 90% of the lesions measured 2 cm was statistically significantly associated with a higher risk of recurrence (P = 0.03). The recurrence-free survival of the nine patients who did not have the planned VRT because of more advanced disease was statistically significantly less (P = 0.003). CONCLUSION: VRT is an oncologically safe procedure in well-selected patients with early-stage disease. Lesion size >2 cm appears to be associated with a higher risk of recurrence. The morbidity of the procedure is low and it allows fertility preservation. PMID- 15350350 TI - Laparoscopic restaging of early stage invasive adnexal tumors: a 10-year experience. AB - Surgical staging of apparent early stage adnexal carcinoma provides indispensable information. A significant number of patients are referred to tertiary centers with inadequate staging information. We report on our experience with late results of laparoscopic restaging procedure in uncompletely managed early adnexal carcinomas. MATERIALS AND METHODS: From 1991 to 2001, 53 laparoscopic restaging operations were performed: 42 patients were restaged early after initial surgery for an ovarian carcinoma (OC) in 35 of them, and for fallopian tube carcinomas (FTCs) in 7 others. Eleven patients were assessed as a second-look procedure, after six courses of platinum-based chemotherapy indicated for a high-risk tumor. The procedure systematically followed the guidelines of laparotomy. RESULTS: All except one (adhesions) procedures were successfully completed. Operative room time averaged 238 min and hospital stay 3.1 days. Only one major complication required laparotomy (1.8%). In the primary restaging group, eight patients were upstaged (19%) and were given chemotherapy. After a 54-month median follow-up, 3 out of the 34 remaining patients diagnosed as stage IA grades 1-2 (6.4%) recurred and died. In the group of 11 second-look operations, 4 were found positive after chemotherapy. One of the positive patient recurred and died. CONCLUSIONS: Laparoscopy seems to be an acceptable technical option to perform restaging of apparently early adnexal carcinomas. It spares the patients the discomfort of repeat laparotomy. Long-term outcome results suggest that laparoscopic staging, provided it meets the standards, accurately detects the patients who need chemotherapy and safely select the patients who can be proposed surgery only. PMID- 15350351 TI - VEGF, VEGFRs expressions and activated STATs in ovarian epithelial carcinoma. AB - OBJECTIVE: To investigate the expressions of vascular endothelial growth factor (VEGF), VEGFRs and activation of signal transducers and activators of transcription (STATs) in ovarian epithelial carcinoma and the relationships among them. METHODS: The tissue samples of 42 primary ovarian epithelial carcinoma, 29 benign ovarian tumor and 11 normal ovarian tissue were used to determine the expression of VEGF, VEGFR1, VEGFR2, P-STAT1, P-STAT3, P-STAT5 and P-STAT6 proteins by immunohistochemical staining. RESULTS: VEGF in ovarian carcinomas was significantly higher than that in benign and normal ovarian tissues. VEGFRs expression was in agreement with VEGF expression. In tumor cells and endothelial cells of ovarian carcinomas, expressions of P-STAT3 and P-STAT5 were significantly higher than those in benign and normal ovarian tissues. In endothelial cells, the expression of VEGFR1 and P-STAT5 closely correlated with each other, as well as VEGFR2 and P-STAT3. However, in ovarian carcinoma cells, expressions of VEGF, VEGFR1 and VEGFR2 were significantly correlated with P-STAT3 and P-STAT5, but not with P-STAT1 and P-STAT6. CONCLUSIONS: There exist overexpressions of VEGF, VEGFRs, and STAT3, STAT5 activation. Furthermore, these results indicate that VEGF secreted by ovarian carcinoma cells may activate STAT pathway via VEGFRs in ovarian carcinoma themselves. PMID- 15350352 TI - Prognostic significance of Bax, Bcl-2, and p53 expressions in cervical squamous cell carcinoma treated by radiotherapy. AB - OBJECTIVE: The objective was to determine the prognostic significance of the immunohistochemical expressions of Bax, Bcl-2, and p53 in squamous cell carcinoma of the uterine cervix treated by radiotherapy alone. METHODS: One hundred and seventy-four cases of squamous cell carcinoma of the cervix (stage Ib-IVa) diagnosed between January 1996 and December 1998 were investigated for Bax, Bcl 2, and p53 expressions and were correlated to the patients' survival. RESULTS: The mean age of the patients was 54.1 with a range of 29-82 years. There were 23 cases with stage I (13.2%), 99 stage II (56.9%), 51 stage III (29.3%), and 1 case with stage IV (0.6%). The 5-year disease-free survival (DFS) was 70.50% and overall survival (OS) was 65.95%. Bax, Bcl-2, and p53 expressions were seen in 68.4%, 25.9%, and 77.6% of cases, respectively. In multivariate analysis by Cox's regression, age, stage, Bax, and Bcl-2 expressions appeared to be independent prognostic predictors of DFS. Bax expression was associated with good survival (hazard ratio, 0.47) while Bcl-2 expression was associated with poor survival (HR, 2.51). In addition, a combination of Bcl-2+/Bax+ was significantly associated with poorer DFS compared to Bcl-2-/Bax+ (HR 3.55). However, none of the markers or combinations was associated with OS. CONCLUSIONS: Evaluation of Bax and Bcl-2 expressions and their co-expression provide independent prognostic information for the clinical course of the disease and therefore could be developed as a prognostic indicator for cervical cancer. PMID- 15350353 TI - A novel retinoid, 4-[3,5-bis (trimethylsilyl) benzamido] benzoic acid (TAC-101), induces apoptosis of human ovarian carcinoma cells and shows potential as a new antitumor agent for clear cell adenocarcinoma. AB - OBJECTIVES: A novel retinobenzoic acid derivative, 4-[3,5-bis (trimethylsilyl) benzamido] benzoic acid (TAC-101), was reported to suppress the growth and invasion of human gastric cancer or hepatocellular carcinoma by induction of apoptosis. We examined the antitumor activity of TAC-101 against human ovarian carcinoma cell lines. METHODS: Apoptosis of human epithelial ovarian carcinoma derived cell lines (RMG-I, RMG-II, RTSG, RMUG-S, RMUG-L, and KF) was investigated by detecting DNA laddering and was quantified by an enzyme-linked immunosorbent assay. Inhibition of apoptosis was also examined using a caspase inhibitor. Furthermore, TAC-101 (8 mg kg(-1) day(-1) orally for 30 days) was investigated in nude mice with subcutaneous RMG-II tumors. A prominent apoptotic response to TAC 101 was observed. The antitumor effects of cisplatin (7 mg/kg intravenously on day 1) and paclitaxel (36 mg/kg intravenously on days 1 and 5) were also assessed for comparison. RESULTS: Apoptosis occurred in all of the cell lines (except KF) in a concentration-dependent manner after exposure to TAC-101 and was markedly induced in RMG-I and RMG-II cells (derived from ovarian clear cell adenocarcinomas). A caspase inhibitor blocked the induction of apoptosis by TAC 101. The maximum inhibition of RMG-II tumor growth in nude mice by TAC-101, cisplatin, and paclitaxel was 45%, 34%, and 47%, respectively. CONCLUSION: Oral TAC-101 shows potential as a novel antitumor agent for ovarian carcinoma, especially ovarian clear cell adenocarcinoma. PMID- 15350354 TI - Improved optimal cytoreduction rates for stages IIIC and IV epithelial ovarian, fallopian tube, and primary peritoneal cancer: a change in surgical approach. AB - OBJECTIVE: To determine the impact of the incorporation of extensive upper abdominal debulking procedures on the rates of optimal primary cytoreduction and complications in stages IIIC and IV epithelial ovarian, fallopian tube, and primary peritoneal carcinomas. METHODS: Two groups of patients were identified for comparison. Group 1, the control group, consisted of 70 consecutive patients who underwent "standard" primary cytoreductive surgery before May 2000. Group 2, the study group, was composed of 70 consecutive patients who underwent surgery after January 2001, during which time, a more comprehensive debulking of upper abdominal disease was used, including diaphragm stripping/resection, splenectomy, distal pancreatectomy, liver resection, resection of porta hepatis tumor, and cholecystectomy. RESULTS: The median age of the entire cohort was 60 years (range, 36-88 years). The majority had stage IIIC disease (86%) and serous histology (76%). Optimal cytoreduction (residual disease /=8 mm) followed by hysteroscopy and endometrial histological findings. Performance characteristics were calculated with correlation to the endometrial histological findings. RESULTS: The mean endometrial thickness was 14.6 +/- 6.2 mm, and the mean intrauterine mass diameter detected by SIS was 11.6 +/- 10.4 mm. There was a gradual decrease in sensitivity and gradual increase in specificity of the SIS studies with the increase in intrauterine mass diameter. False-negative and false positive of SIS were 2.4% and 8.2%, respectively. ROC curve analysis of intrauterine mass revealed 5 mm as the best accurate cutoff value for the diagnosis of endometrial pathologies, with a sensitivity of 74.1%, specificity of 93.0%, and positive predictive value of 88.3% and negative predictive value of 84.2%. The risk of endometrial pathology was elevated by 1.37-fold, with any additional millimeter of diameter of the intrauterine mass. The mean diameter of the intrauterine mass gradually increased the greater the severity of the histological findings. CONCLUSIONS: Sonohysterography improves the accuracy of diagnosis of intrauterine mass in asymptomatic postmenopausal tamoxifen-treated patients. The size of the intrauterine mass correlates with the severity of the endometrial pathology. PMID- 15350370 TI - Anticancer effects of (-)-epigallocatechin-3-gallate on ovarian carcinoma cell lines. AB - PURPOSE: A constituent of green tea, (-)-epigallocatechin-3-gallate (EGCG), has been known to possess anti-cancer properties. In this study, we investigated the time-course anticancer effects of EGCG on human ovarian cancer cells to provide insights into the molecular-level understanding of growth suppression mechanism involved in EGCG-mediated apoptosis and cell cycle arrest. METHODS: Three human ovarian cancer cell lines (p53 negative, SKOV-3 cells; mutant type p53, OVCAR-3 cells; and wild type p53, PA-1 cells) were used. The effect of EGCG treatment was studied via cell count assay, cell cycle analysis, FACS, Western blot, and macroarray assay. RESULTS: EGCG exerts a significant role in suppressing ovarian cancer cell growth. Also, EGCG showed growth inhibitory effects in each cell line in a dose-dependent fashion and induced apoptosis and cell cycle arrest. The cell cycle was arrested at the G(1) phase by EGCG in SKOV-3 and OVCAR-3 cells. In contrast, the cell cycle was arrested in the G(1)/S phase arrest in PA-1 cells. EGCG differentially regulated the expression of genes and proteins (Bax, p21, Retinoblastoma, cyclin D1, CDK4, Bcl-X(L)) more than 2-fold, showing a possible gene regulatory role of EGCG. The continual expression in p21WAF1 suggests that EGCG acts in the same way with p53 proteins to facilitate apoptosis after EGCG treatment. And Bax, PCNA, and Bcl-X are important in EGCG-mediated apoptosis. In contrast, CDK4 and Rb are not important in ovarian cancer cell growth inhibition. CONCLUSION: EGCG can inhibit ovarian cancer cell growth through induction of apoptosis and cell cycle arrest as well as regulation of cell cycle-related proteins. Thereby, the EGCG-mediated apoptosis can be applied to an advanced strategy in the development of a potential drug against ovarian cancer. PMID- 15350371 TI - The anatomy of complications workshop: an educational strategy to improve the training and performance of fellows in gynecologic oncology. AB - OBJECTIVES: In Australasia, the USA, and Canada, certified gynecologic oncologists (CGO) are expected to be competent in performing intestinal, urological, and vascular procedures relevant to gynecologic oncology. This competency may be difficult to achieve and experience with simulation rather than human patients may be preferable during initial training. We present the structure, objectives, rationale, and outcomes for a surgical skill workshop aimed to improve the performance of fellows in gynecologic oncology. WORKSHOP STRUCTURE: The overall objective of the workshop is the prevention and management of complications in gynecologic oncology surgery. Preworkshop preparation includes anatomical illustrations to guide reading and a training video describing surgical procedures and the relevant aspects of ovine and porcine anatomy. There are four modules: (i) Anatomy includes an interactive lecture, cadaveric dissection, and examination of prosections. (ii) Porcine surgery involves the demonstration, practice, and learning of techniques needed to deal with unexpected or deliberate operative injury to bowel, bladder, ureter, and major blood vessels and performance of less common operations. (iii) Live ovine surgery allows further supervised practice of these procedures plus the repair of vascular injury. (iv) Case presentations allow each participant to present a complicated case in a facilitated group session with discussion and feedback from their peers. This session is controlled, nonthreatening, and interactive. CONCLUSION: Participant feedback and evaluation suggests that this workshop format and the opportunity to practice less common procedures are helpful to clinical practice. This educational model is presented in the hope that it may act as a template for the development of similar programs for fellows in other countries. PMID- 15350372 TI - Clinical experience with combination paclitaxel and carboplatin therapy for advanced or recurrent carcinosarcoma of the uterus. AB - OBJECTIVE: The purpose of the study was to evaluate the efficacy of combination chemotherapy with paclitaxel and carboplatin in patients with advanced or recurrent carcinosarcoma of the uterus. METHODS: A retrospective review was carried out at Miyagi Prefecture Cancer Research Center Hospital. Six patients pathologically diagnosed with uterine carcinosarcoma were treated with paclitaxel (175 mg/m(2) given intravenously over 3 h) and carboplatin (dosed at AUC 6) every 3 weeks at our center between 1997 and 2003. Responses and adverse effects were assessed according to Response Evaluation Criteria in Solid Tumors and National Cancer Institute-Common Toxic Criteria, respectively. RESULTS: All six patients were evaluable for toxicity, and no unacceptably severe toxicities were reported. Grades 3 and 4 hematologic toxicities occurred, but all of them were overcome by adequate treatment with granulocyte colony-stimulating factor and blood transfusions. Five of six patients had measurable disease and thus were evaluable for response: Four patients had a complete response (CR) and the remaining patient had progressive disease (PD). The median progression-free interval (PFI) for all six cases was 18 months, with a median overall survival of 25 months. CONCLUSIONS: Although the number of cases was small, the regimen evaluated in the current study demonstrated higher activity and lesser toxicity than those found in previous studies in patients with advanced or recurrent uterine carcinosarcoma. Additional phase II clinical studies are necessary to evaluate fully the benefits of this regimen. PMID- 15350373 TI - Frozen section examination of pelvic lymph nodes in endometrial and cervical cancer: accuracy in patients submitted to neoadjuvant treatments. AB - OBJECTIVE: The aim of the study was to analyze the accuracy, the positive and negative predictive values, the specificity, and the sensitivity of the pelvic node frozen section examination in endometrial (EC) and cervical cancer (CC) patients. An accurate evaluation of the results of the frozen section examination in patients submitted to neoadjuvant treatments (chemotherapy and radiochemotherapy), and a comparison between specialist and general pathologist results were performed. METHODS: A total of 186 consecutive patients with endometrial [52 cases (27.9%)] and cervical [134 cases (72.1%)] cancer underwent surgery at our Department between January 2000 and September 2003. All patients underwent a systematic pelvic lymphadenectomy. While all definitive diagnosis were primarily performed or secondarily revised by the specialist pathologist, the frozen section examination was performed in 65 (35%) cases by the specialist pathologist and in 121 (65%) cases by a general pathologist. RESULTS: Two thousand seven hundred eighteen out of a total of 6710 pelvic lymph nodes, which underwent a definitive histological analysis, were also analyzed at intraoperative frozen section examination. In our series, we observed 10 false negative and five false positive cases. Six out of the 10 (60%) false negative cases and two out of the five (40%) false positive were found in the neoadjuvant treated cervical cancer patients. In this subgroup, five out the six (83.3%) false negative and the two false positive diagnoses were made by the general pathologist, while the specialist pathologist registered only one false negative diagnosis. CONCLUSION: Intraoperative examination of pelvic lymph nodes during surgery for endometrial and cervical cancer can be considered a safe procedure in the presence of an expert gynecologic oncological team, and can be safely performed in patients submitted to neoadjuvant treatments. PMID- 15350374 TI - AKT involvement in cisplatin chemoresistance of human uterine cancer cells. AB - OBJECTIVE: The aim of this study was to investigate the possible involvement of Akt activity and specific isoforms (Akt1, Akt2, and Akt3) in the resistance of human uterine cancer cells to cisplatin. METHODS: Two different endometrial (HEC 1-A and KLE) and one cervical (HeLa) cancer cell lines all known as wild-type PTEN (tumor suppressor phosphatase tensin homologue, a lipid phosphatase involved in the negative regulation of Akt activity) were used for these studies. RESULTS: Basal levels of Akt1, Akt2, and Akt3 mRNAs were determined by real-time quantitative RT-PCR studies and Western blot analyses were carried out to determine protein abundance of each isoforms. Akt1 mRNA and protein were present in all cell lines studied. Akt2 and Akt3 mRNAs and proteins were strongly expressed in KLE cells. Surprisingly, Akt phosphorylation was found in KLE expressing high levels of wild-type PTEN protein. KLE cells remained resistant to PI 3-K inhibitor, indicating that Akt phosphorylation might be, in part, independent of PI 3-K in this cell line. Cisplatin induced apoptosis in HeLa and HEC-1-A cells, but KLE cells expressing Akt2 and Akt3 remained more resistant to cisplatin. Knockout of Akt isoforms using specific siRNA technology increased the sensitivity of KLE cells toward cisplatin and caused a significant induction of cell death. CONCLUSION: Taken together, these results suggest that specific Akt isoforms such as Akt2 and Akt3 might be involved in chemoresistance to cisplatin and that these isoforms could be putative targets for gene therapy in uterine cancers. PMID- 15350375 TI - Analysis of the time interval between diagnoses in women with double primary breast and ovarian or primary peritoneal cancers. AB - OBJECTIVES: The time interval between diagnoses of breast and epithelial ovarian cancer is not well established in women with dual primary tumors of both organ sites. Our goals were to examine the time interval between diagnoses and identify any relationship to clinicopathologic factors. METHODS: We identified 49 patients who developed both cancers. These patients were divided into two groups: group 1 patients developed breast cancer first, group 2 patients had ovarian before breast cancer. The risk of a BRCA1 or BRCA2 mutation in our study subjects was estimated using the BRCAPRO. Parameters were compared using either the chi(2) or the Kruskal-Wallace test. RESULTS: There were 26 and 23 patients in groups 1 and 2, respectively. The mean time interval was longer in group 2 (86 vs. 45 months; P = 0.013). Median PFS and OS were longer in group 2 for both cancers [PFS: 161 vs. 61 months for breast (P = 0.85) and 132 vs. 39 months for ovarian (P = 0.019); OS: 250 vs. 115 months for breast (P = 0.77) and 277 vs. 42 months for ovarian (P = 0.0013)]. OS was longer in group 2 for both cancers combined 217 vs. 115 (P = 0.026). The estimated risk of BRCA mutation was at least 20% in the majority of the patents. CONCLUSIONS: Our data indicates that the time interval between the diagnosis of breast and ovarian carcinomas is 4 years. In contrast, the time interval between the diagnosis of ovarian and breast carcinomas is 7 years. These results could be useful in counseling women at risk. PMID- 15350376 TI - Comparative analysis of characteristics of women with cervical cancer in high- versus low-incidence regions. AB - OBJECTIVE: To identify particular characteristics that might help explain the markedly higher rate of invasive cervical cancer in southern China as compared with Australia. METHODS: One hundred eighty-five women with cervical cancer were recruited between 1999 and 2001: 106 from Guangzhou and Changsha (southern China), and 79 from Sydney (southeast Australia). Demographic and risk factor information was obtained by questionnaire; clinicopathologic data were extracted from hospital records. The human papillomavirus (HPV) status of cancer biopsies was ascertained by consensus PCR assays, direct sequencing and/or Amplicor trade mark hybridisation. RESULTS: The mean age of the Chinese was significantly lower than the Australians (44 versus 53 years), although mean age at first sexual intercourse was similar. Australian women were more likely to smoke, to report multiple sexual partners and to have a history of sexually transmitted diseases (but not of genital warts). However, they were better educated, were more frequent users of barrier contraception and were far more likely to report regular Pap smears before diagnosis. The HPV positivity rate of Chinese cancers (83%) was less than Australian tumours (90%); but HPV 16 and 18 were the most common genotypes in both populations (59% and 77%), and predominated in cancers from younger women. HPV types 58 or 59 were amplified from 12 (14%) of the Chinese but from only one Australian cancer. CONCLUSIONS: Cervical cancer is not only more common in China but also develops at a younger age than in Australia. While significant differences in some risk factors were observed, the much lower participation in cervical screening in southern China is likely to be of greatest consequence. PMID- 15350377 TI - Non-Hodgkin's lymphoma presenting as a pelvic mass with elevated CA-125. AB - BACKGROUND: We report a case of pelvic lymphoma with an elevated serum CA-125 level, initially misdiagnosed as ovarian carcinoma. A review of the literature is presented and a possible mechanism for CA-125 elevation in diseases other than ovarian cancer is discussed. CASE: A 50-year-old woman presented with symptoms of progressive dyspnea, early satiety, fatigue, and weight loss. Workup revealed a pelvic mass and an elevated CA-125 level. Paclitaxel and carboplatin were administered to facilitate therapy and provide symptomatic relief for a presumed bulky ovarian carcinoma. A biopsy was obtained after the initiation of chemotherapy, yielding the diagnosis of diffuse large B cell non-Hodgkin's lymphoma, stage II-B. A regimen of cyclophosphamide, doxorubicin, vincristine, and prednisone followed by radiotherapy resulted in long-term disease remission. A search of the literature revealed several clinical series describing the elevation of CA-125 in a variety of diseases, both benign and malignant. CONCLUSIONS: In the setting of a newly diagnosed pelvic mass, care should be taken when interpreting an elevated CA-125 level. While ovarian cancer is high on the list of differential diagnoses, lymphoma cannot be excluded until a tissue diagnosis is obtained. PMID- 15350378 TI - Conversion of an incontinent urinary conduit to a continent urinary reservoir (Miami Pouch). AB - OBJECTIVE: External urinary or gastrointestinal appliances can impair a patient's quality of life. We report on the feasibility of converting an incontinent colonic urinary diversion to a continent urinary reservoir (Miami Pouch). CASE: We describe the case of a 66-year-old white female with a history of stage Ib(2) cervical cancer treated by radical abdominal hysterectomy and adjuvant radiation therapy. The patient developed severe radiation cystitis with a neurogenic bladder and bilateral ureteral obstruction. After failing conservative management, a urinary diversion with a transverse colon conduit was performed. The patient remained without evidence of disease for 2 years and led an active lifestyle with regular tennis games. After 7 months of an external appliance for the urinary conduit, the patient presented to the University of Miami for conversion to a continent urinary mechanism which would not require an appliance. We performed an exploratory laparotomy, conversion of a transverse colon conduit to a continent ileo-colonic urinary reservoir (Miami Pouch). There were no postoperative complications. The patient remains disease-free and performs self catheterization with no need for an external appliance. The patient has been able to resume an active life including sports. CONCLUSIONS: Successful conversion of an incontinent urinary conduit to a continent urinary reservoir is possible in a select case resulting in a perceived improvement of quality of life. PMID- 15350379 TI - Uterine smooth-muscle tumor of uncertain malignant potential metastasizing to the humerus as a high-grade leiomyosarcoma. AB - BACKGROUND: Smooth-muscle tumors of uncertain malignant potential represent a variant of uterine leiomyoma and remain a dilemma due to their uncertain clinical behavior. Most of these tumors have a benign clinical course, and coagulative tumor cell necrosis has usually been associated with worse outcome. CASE: We present a case of a uterine smooth-muscle tumor (SMT) of uncertain malignant potential without coagulative tumor cell necrosis that has metastasized to the humerus. The metastatic lesion was a high-grade leiomyosarcoma consistent with uterine origin. CONCLUSIONS: Smooth-muscle tumors of uncertain malignant potential that demonstrate a lack of coagulative tumor cell necrosis do not necessarily insure benign clinical outcome. These lesions can recur and metastasize to distant sites, thus requiring long-term follow-up. PMID- 15350380 TI - Ovarian hemangioma presenting with hyperandrogenism and endometrial cancer: a case report. AB - BACKGROUND: Hemangiomas are very rare tumors of the ovary. Here, we report a case of a mixed capillary and cavernous ovarian hemangioma and endometrial carcinoma presenting with postmenopausal bleeding, male pattern receding frontal hairline, and high serum androgen and estradiol levels. CASE: A 70-year-old White female underwent laparotomy for endometrial carcinoma. Intraoperative frozen-section examination of the uterus revealed a 3.5 x 3 cm, grade 1 endometrioid adenocarcinoma of the endometrium with more than 50% myometrial invasion. The left ovary contained a 1.5 x 1 x 1 cm, well-circumscribed hemorrhagic nodule on the cut surface. Final histopathological examination of the small nodule demonstrated multiple, enlarged, blood-filled vascular channels lined by a single layer of flattened regular endothelial cells with no atypical features. Vascular spaces within the tumor were of different sizes, ranging from small to large, and were separated by connective tissue. The surrounding ovarian stroma was hyperplastic and contained clusters of luteinized stromal cells. Microscopy of the right ovary showed minimal stromal proliferation and no luteinization of the ovarian stroma. CONCLUSION: This is the first case of an ovarian hemangioma synchronous with a well-differentiated endometrial carcinoma. Absence of estrogen and progesterone receptors in the endothelial cells of the hemangioma suggests that ovarian hemangiomas may occur independent of stimulation by estrogen and progesterone. PMID- 15350381 TI - Squamous cell carcinoma of the cervix presenting as lymphangitic carcinomatosis: a case report and review of the literature. AB - BACKGROUND: Lymphangitic carcinomatosis (LC) secondary to carcinoma of the cervix is rare. The presenting symptoms are misleading and nonspecific, which often leads to delayed diagnosis. CASE: We present the case of a 24-year-old woman with dyspnea and lower back pain, who was found to have stage IIIb squamous cell carcinoma of the cervix complicated by pulmonary LC. CONCLUSIONS: Pulmonary LC is a rare but important manifestation of metastatic cervical cancer. Patients present with severe respiratory compromise, which mimics other, more common disease states. Diagnosis can be achieved by transbronchial biopsy. Optimal treatment of the patient with LC is not well defined, but a trial of chemotherapy and/or intravenous steroids may be warranted. Prognosis for this condition is poor. PMID- 15350382 TI - Primary lung large cell carcinoma metastatic to the vulva: a case report and review of the literature. AB - BACKGROUND: Metastatic cancer to the vulva is a rare diagnosis and is estimated to account for 5-8% of all vulvar cancers. CASE: A 57-year-old postmenopausal woman was referred for the evaluation of a new vulvar mass. CT scan of the pelvis demonstrated a 4 x 6 cm lobular mass of the left labia. An outpatient excisional biopsy revealed a poorly differentiated large cell carcinoma with prominent necrosis and focal perineural invasion consistent with a bronchogenic carcinoma. A subsequent PET scan of the chest revealed a large primary lung carcinoma confirmed by CT-guided biopsy that was identical to the vulvar tumor. CONCLUSION: This case represents the only literature described primary lung carcinoma presenting as a vulvar metastasis. PMID- 15350383 TI - Anastrozole, an aromatase inhibitor, and medroxyprogesterone acetate therapy in premenopausal obese women with endometrial cancer: a report of two cases successfully treated without hysterectomy. AB - BACKGROUND: Hormonal therapy for endometrial cancer is occasionally warranted in the premenopausal woman who is interested in maintaining fertility. Combining progesterone with an agent that eliminates the adipose production of estrogen will theoretically be more effective than progesterone alone. CASES: Two cases of reproductive-aged women with grade 1 endometrial cancer who were treated with medroxyprogesterone acetate and anastrozole daily for 3 and 6 months subsequently reverted to normal endometrium. CONCLUSION: Progesterone combined with the elimination of adipose production of estrogen may be an effective therapy in well differentiated endometrial cancer in the obese premenopausal woman. PMID- 15350384 TI - Struma Ovarii with a focus of papillary thyroid cancer: a case report and review of the literature. AB - BACKGROUND: Malignant Struma Ovarii is a rare ovarian neoplasm that is usually asymptomatic and infrequently diagnosed preoperatively. A few case studies have described associated thyrotoxicosis. CASE: A 46-year-old female presented for her annual gynecologic exam during which a pelvic mass was appreciated on physical examination. Patient was asymptomatic at presentation. A follow-up ultrasound confirmed the presence of a 16-cm mass in the right adnexa. Patient underwent a total abdominal hysterectomy with bilateral salpingo-oophorectomy. Histopathology revealed a mature cystic teratoma with features of Struma Ovarii, containing a single 5-mm focus of papillary cancer within the thyroid tissue. Patient subsequently had a thyroid scan that was normal with normal thyroid function. There was no evidence of metastasis. CONCLUSION: Malignant Struma Ovarii is a very rare disease and there are various approaches to treatment based on staging. In our review, we found a higher rate of metastasis than previously reported. Recurrence was seen after an average period of 4 years. We recommend follow-up with surveillance thyroglobulin levels in cases of malignant Struma Ovarii for at least 10 years. PMID- 15350385 TI - Primary leiomyosarcoma of the vagina: a case report involving a TVT allograft. AB - INTRODUCTION: Primary malignant lesions of the vagina represent less than 2% of all gynecologic malignancies. Primary vaginal sarcomas account for about 2% of all malignant vaginal lesions, with leiomyosarcoma being the most common vaginal sarcoma found in adult women. CASE: We report a case of primary vaginal leiomyosarcoma occurring in the field of a prior tension-free vaginal tape (TVT) procedure using a Bard Duraderm allograft. CONCLUSION: This report represents the first report, to our knowledge, of a vaginal sarcoma arising in the field of a Bard Duraderm TVT allograft. Although the product is no longer available for this use, the ongoing studies of the TVT procedure and outcomes should include this potential complication in their review. PMID- 15350386 TI - Contralateral groin recurrence in patients with stage I Bartholin's gland squamous cell carcinoma and negative ipsilateral nodes: report on two cases and implications for lymphadenectomy. AB - BACKGROUND: Modalities of inguinal lymphadenectomy in patients with early stage Bartholin's gland squamous cell carcinoma (BGC) remain discussed. The aim of this paper is to report 2 patients with stage 1 BGC and negative ipsilateral groin nodes who developed a contralateral groin recurrence. CASES: Two patients treated for BGC and undergoing an ipsilateral groin nodes dissection (with absence of nodal involvement) presented contralateral groin recurrence. These recurrences were observed 14 and 3 months after the initial treatment in the first and second case. In the first case, the nodal recurrence was associated with a local vaginal recurrence. CONCLUSIONS: These 2 cases suggest that << cross >> groin nodal involvement can occur, even in patients with early stage BGC. Bilateral groin dissection should be performed in patients with BGC. PMID- 15350387 TI - Ovarian metastasis from primary renal cell carcinoma: report of a case and review of literature. AB - BACKGROUND: Ovarian metastasis from renal cell adenocarcinoma is quite rare. They are often mistaken for primary clear cell tumours of the ovary because of histological similarity and propensity to appear years before or after the renal primary is detected. CASE: A case of ovarian metastasis from renal cell adenocarcinoma is reported. Patient developed bilateral ovarian metastasis 7 years after diagnosis of renal primary. She is alive 2 years after removal of ovarian tumours, with evidence of metastases to pelvic and paraaortic lymph nodes. Review of literature with analysis of 11 such reported cases in English literature is included. CONCLUSION: Although rare, possibility of metastasis from renal cell adenocarcinoma should be considered as a differential diagnosis in cases of clear cell ovarian tumours. Imaging studies are warranted in such cases to rule out renal tumour. It is important to differentiate between the two because of therapeutic and prognostic implications. Careful histological and histochemical analysis may help to differentiate between the two. PMID- 15350388 TI - Atypical sigmoid metastasis from a high-grade mixed adenocarcinoma of the ovary. AB - BACKGROUND: Intraperitoneal seeding is the most common form of dissemination of epithelial ovarian cancer. Metastasis to the bowel mucosa can occur by invasion from the serosal surface or infiltration of the submucosal capillary network. Hematogenous dissemination usually occurs in the presence of advanced peritoneal disease. CASE: A 39-year-old gravida 3 para 2 woman was diagnosed in October 2000 with a large pelvic mass. She underwent an exploratory laparotomy and a left salpingo-oophorectomy with multiple abdominal and pelvic biopsies. She was diagnosed with an ovarian tumor of low malignant potential, and no further treatment was recommended. Six months later, she developed abdominal discomfort and constipation. A colonoscopy was performed, and a biopsy showed metastatic carcinoma of ovarian origin. The patient presented to The University of Texas M.D. Anderson Cancer Center in September 2001 for consultation. The surgical pathology evaluation from her previous surgery indicated high-grade ovarian carcinoma. The patient underwent a total abdominal hysterectomy, right salpingo oophorectomy, infracolic omentectomy, right pelvic lymph node sampling, and segmental resection with primary end-to-end sigmoid colon anastomosis. The tumor within the colon was a polypoid mass arising from the mucosa with no involvement of the colonic wall. Microscopically, the tumor was a high-grade ovarian papillary serous carcinoma with areas of endometrioid adenocarcinoma. The colonic tumor was immunohistochemically positive for cytokeratin-7 and negative for cytokeratin-20. The patient was treated with six cycles of carboplatin and paclitaxel. The patient then incidentally developed disseminated sarcoidosis. At the time of this report, the patient had no evidence of recurrent or metastatic disease for 2 years. CONCLUSIONS: Epithelial ovarian carcinomas may recur as intraluminal bowel lesions with serosal sparing even in the absence of peritoneal disease. Immunohistochemical staining using cytokeratins-7 and -20 may prove useful in differentiating such lesions from primary colonic malignancies. PMID- 15350389 TI - Excellent progression-free survival with liposomal doxorubicin for a patient with recurrent ovarian malignant mixed mullerian tumor: case report and literature review. AB - INTRODUCTION: Ovarian malignant mixed mullerian tumor (MMMT) is a rare, highly aggressive, fatal disease. Patients have a median survival of 18 months and a 5 year survival rate of only 8%. Optimal cytoreduction surgery plus platinum-based combination chemotherapy are associated with better outcomes. CASE REPORT: A 65 year-old patient of stage IIIc ovarian MMMT having obtained a 41-month remission after four courses of aggressive surgical debulking procedures, platinum containing chemotherapy, and intraoperative radiotherapy suffered from multi focal recurrences and obtained another 22-month progression-free survival after treatment with monthly liposomal doxorubicin (Lipo-Dox) for 14 courses and Lipo Dox/carboplatin for subsequent 6 courses without obvious toxicity. DISCUSSION: Liposomal doxorubicin might be useful as salvage chemotherapy for heavily pretreated, recurrent ovarian MMMT. A prospective trial is needed for more proof. PMID- 15350390 TI - Breast cancer metastatic to the vulva. AB - BACKGROUND: Primary cancer of the vulva is not common, constituting only 3-5% of all gynecologic malignancies and <1% of all cancer in the female. Metastatic tumors of the vulva are even more unusual, constituting only 5-8% of all vulvar cancers. CASE: A 32-year-old female underwent total mastectomy with axillary lymph node dissection for left breast cancer. The patient was well with a postoperative follow-up period of 40 months till a 1.2-cm lump was noted in her left labium majus. Excisional biopsy was performed and histologic examination was done. Pathologic examination of the breast cancer revealed coexistence of intraductal carcinoma and invasive lobular carcinoma. The histologic feature of the invasive lobular carcinoma was consistent with that of the vulvar lesion. The vulvar cancer was consistent with a metastatic lobular carcinoma from the breast. CONCLUSION: Only with careful gynecologic surveillance in women with breast cancer can the unusual sites of metastasis be detected earlier and appropriately treated. PMID- 15350391 TI - Vaginal cuff recurrence of endometrial cancer treated by laparoscopic-assisted vaginal hysterectomy. PMID- 15350393 TI - Relevance of the sentinel node procedure in endometrial cancer. PMID- 15350395 TI - Hepatic oval cells: helping redefine a paradigm in stem cell biology. PMID- 15350396 TI - Meiotic DNA replication. PMID- 15350397 TI - Pollen tube guidance: the role of adhesion and chemotropic molecules. PMID- 15350398 TI - The biology and diagnostic applications of fetal DNA and RNA in maternal plasma. PMID- 15350399 TI - Advances in tissue engineering. PMID- 15350400 TI - Directions in cell migration along the rostral migratory stream: the pathway for migration in the brain. PMID- 15350401 TI - Retinoids in lung development and regeneration. PMID- 15350402 TI - Structural organization and functions of the nucleus in development, aging, and disease. PMID- 15350404 TI - Sources, distribution and behaviour of methyl tert-butyl ether (MTBE) in the Tamar Estuary, UK. AB - Negligible information is currently available concerning levels of the fuel additive methyl tert-butyl ether (MTBE) in European estuaries or coastal environments. MTBE was measured at selected potentially contaminated harbours and marinas, and throughout an axial transect of the Tamar Estuary, UK. Headspace solid-phase microextraction was used in combination with GC-MS for its determination. MTBE was detected in water samples from all stations at concentrations ranging from a few ng/l to a maximum of 194 ng/l (in a semi enclosed harbour). Elevated levels were generally associated with motor vehicle and boating activities. The Tamar road and rail bridges provided a major input to the lower estuary, downstream of which conservative mixing appeared depressed, probably through volatilisation and possibly through salting-out. The selected analytical system (using a Carboxen/PDMS fiber) proved both rapid and highly sensitive (with a detection limit of 6 ng/l). During method development, salinity was shown to have a major influence in controlling the extraction efficiency and it was found necessary to adjust salinity in all samples (to 75% saturation) prior to extraction. From these tests, we (for the first time) estimated the Setschenow ("salting-out") constant of MTBE to be 0.11 l/mol. PMID- 15350405 TI - Evaluation of solubilizing ability of humic aggregate basing on the phase separation model. AB - Solubilizing abilities of aggregates of humic acid (HA) to chlorinated benzenes (CBs) were investigated by means of the apparent water solubility enhancement. Both the water solubilities of 1,4-dichlorobenzene (DCB) and 1,2,4,5 tetrachlorobenzene (TeCB) linearly increased with increasing concentration of HA above the critical micelle concentration (CMC). Such solubilization behavior of CBs for HA was compatible with those for sodium dodecyl sulfate (SDS). These results indicate that the solubilization of CBs in the aqueous solution of HA above the CMC can be interpreted on the basis of the phase-separation model. Thus, the partition coefficients (K(mic)) of CBs between water and HA aggregate phases were calculated by assuming this model. The fact that the K(mic) value increased with increasing K(ow) of CBs supported the partition into the HA aggregate phase by hydrophobic interaction. The estimated K(mic) values of DCB were not dependent on the solution pH. Both K(mic) values of DCB and TeCB for the HA aggregate were found to be 4-5-fold lower than those of SDS. PMID- 15350406 TI - Effect of cadmium on free amino acid, glutathione and ascorbic acid concentrations in two barley genotypes (Hordeum vulgare L.) differing in cadmium tolerance. AB - Hydroponic experiment was carried out to study the effect of three Cd levels on glutathione (GSH), free amino acids (FAA), and ascorbic acid (ASA) concentration in the different tissues of 2 barley cultivars with different Cd tolerance. Cadmium concentration in both roots and shoots increased with external Cd level, while biomass and ASA concentration declined, and Wumaoliuling, a Cd-sensitive genotype was more affected than ZAU 3, a Cd-tolerant genotype. The effect of Cd on GSH concentration was dose- and time-dependent. In the 5 d exposure, root GSH concentration increased in 0.5 microM Cd treatment compared with control, but decreased significantly in 5 microM Cd treatment, irrespective of genotypes. However, in the 10 d exposure, GSH concentration in all plant tissues decreased with increasing Cd levels in the culture medium, and Wumaoliuling was much more affected than ZAU 3. Cadmium treatment greatly altered FAA concentration and composition in plants. The effect of Cd on glutathione (Glu) concentration in roots varied with genotypes. ZAU 3 showed a steady increase in root Glu concentration in both 0.5 and 5 microM Cd treatments, while Wumaoliuling was decreased by 38.0% in 5 microM Cd treatment, compared with the control. The results indicate that GSH and ASA are attributed to Cd tolerance in barley plants, and the relative less reduction in GSH concentration in ZAU 3 under Cd stress relative to the control may account for its higher Cd tolerance. PMID- 15350407 TI - Photodegradation of the herbicide Norflurazon sensitised by Riboflavin. A kinetic and mechanistic study. AB - The kinetics and mechanism of the Riboflavin (Rf)-promoted photochemical degradation with visible light of the herbicide Norflurazon (NF) has been studied by time-resolved and stationary techniques. Using light of wavelength higher than 400 nm--a region where NF is totally transparent--and with concentrations of Rf and NF of ca. 0.02 and 1 mM, respectively, only the excited triplet state of Rf ((3)Rf*) is quenched by NF, in competition with dissolved ground state triplet oxygen, O(2)((3)Sigma(g)(-)). NF degradation mainly occurs by reaction with superoxide radical anion O(2)(-) formed through two electron transfer steps: from NF to (3)Rf*, yielding Rf radical anion, and from this anion to O(2)((3)Sigma(g)( )), regenerating ground state Rf. Although singlet molecular oxygen is also produced, NF only quenches this oxidative species in a physical mode. The global result is the photoprotection of the sensitiser and the photodegradation of NF. PMID- 15350408 TI - Organic matrix in produced water from the Osage-Skiatook petroleum environmental research site, Osage county, Oklahoma. AB - Produced water (water co-produced with oil and gas) constitutes the single largest waste stream for oil and gas industry. Reclaiming this water for beneficial use is thought to be one of the most practical solutions that can solve both environmental and water shortage problems. The feasibility of this practice depends on the ability to remove its chemical content to the levels that meets the appropriate standards. Organic compounds are probably the most difficult fraction to handle. In this paper, the discrete organic compounds and non-volatile, macromolecular organic compounds (i.e., natural organic matter- NOM) of three produced water samples from the Osage-Skiatook Environmental Research site were characterized. Two of the three produced waters had very little contribution from NOM, while one of the samples had about 23% NOM contribution to its organic matrix pool. Fluorescent spectrophotometric scans provided little differentiation among the organic quality of the produced water, while pyrolysis-GC/MS showed that the NOM characteristics of the three produced waters were distinct. Specifically, the overall halogenated content and aromaticity of the NOM were found to be possible qualifiers that distinguish produced water from the coalbed methane well from produced water from the oil well. And the specific chemical fragments that are linked to polysaccharide sources were found to be potential identifiers that distinguish produced water from the newer oil well from produced water from the older oil well. These identifiers were, however, only suggested for this preliminary study. More samples must be included to build a substantial database on produced water NOM to confirm and identify more markers. PMID- 15350409 TI - Adsorption-desorption, persistence and leaching behavior of thifluzamide in alluvial soil. AB - Investigations were undertaken to study the adsorption-desorption, persistence and leaching of thifluzamide (2',6'-dibromo-2-methyl-4'-trifluoromethoxy-4 trifluoro methyl-1,3-thiazole-5-carboxanilide) in an alluvial soil under laboratory conditions. The adsorption-desorption studies were carried out using batch equilibration technique. The results revealed high but weak adsorption of thifluzamide in alluvial soil with K(F) value of 9.62 and 'n' value of 0.63. About 47-62% of the adsorbed amount got desorbed in four desorption cycles, which further substantiate the hypothesis of weak binding. The hysteresis coefficient varied from 0.19 to 0.40. Persistence studies carried out at three concentration levels (0.1, 1.0 and 10.0 microgg(-1)) and under three moisture conditions (air dry, field capacity moisture and submerged) revealed that thifluzamide is a persistent chemical and only 19.5-54.0% dissipation was recorded on 90th day. However, it appears that aerobic microbes are more efficient in degrading thifluzamide than anaerobic microbes. The preliminary leaching studies carried out in the laboratory revealed that thifluzamide was moderately mobile in alluvial soil. Only small amounts (<1%) were recovered from leachate fractions whereas major portion remained in 0-15 cm soil depth. PMID- 15350410 TI - Influence of kinetic sorption and diffusion on pesticide movement through aggregated soils. AB - Laboratory studies were carried out to investigate solute leaching at different times from application in relation to temperature and initial soil moisture. Aggregates of a heavy clay soil were treated with a non-interactive solute (bromide) and the herbicides chlorotoluron, isoproturon and triasulfuron. The soil was incubated at 90% field capacity and either 5 or 15 degrees C. The influence of application to initially dry and initially wet aggregates on the behaviour of isoproturon was also investigated. At intervals, samples were either leached in small columns, centrifuged to characterise the fraction of chemical available in pore water under natural moisture conditions or extracted with organic solvents to assess total residues in soil. Bromide concentrations in leachate and in pore water extracted by centrifugation were constant with time. In contrast, availability for leaching and concentration in pore water of the herbicides decreased with increasing time from application in soil incubated at 15 degrees C. The effect of residence time was much smaller at 5 than at 15 degrees C. At the higher temperature, pesticide concentrations in leachate and pore water declined faster than would be expected from degradation alone, probably due to slow diffusion of the pesticides into soil aggregates where they are less available for leaching and/or slow sorption-desorption. The faster decline in availability for leaching at 15 than at 5 degrees C was attributed to faster degradation of the readily available fraction. There was no significant influence of initial soil moisture on either the leaching behaviour of isoproturon or its availability in soil water. PMID- 15350411 TI - Characterisation of the dilute HCl extraction method for the identification of metal contamination in Antarctic marine sediments. AB - A regional survey of potential contaminants in marine or estuarine sediments is often one of the first steps in a post-disturbance environmental impact assessment. Of the many different chemical extraction or digestion procedures that have been proposed to quantify metal contamination, partial acid extractions are probably the best overall compromise between selectivity, sensitivity, precision, cost and expediency. The extent to which measured metal concentrations relate to the anthropogenic fraction that is bioavailable is contentious, but is one of the desired outcomes of an assessment or prediction of biological impact. As part of a regional survey of metal contamination associated with Australia's past waste management activities in Antarctica, we wanted to identify an acid type and extraction protocol that would allow a reasonable definition of the anthropogenic bioavailable fraction for a large number of samples. From a kinetic study of the 1 M HCl extraction of two Certified Reference Materials (MESS-2 and PACS-2) and two Antarctic marine sediments, we concluded that a 4 h extraction time allows the equilibrium dissolution of relatively labile metal contaminants, but does not favour the extraction of natural geogenic metals. In a regional survey of 88 marine samples from the Casey Station area of East Antarctica, the 4 h extraction procedure correlated best with biological data, and most clearly identified those sediments thought to be contaminated by runoff from abandoned waste disposal sites. Most importantly the 4 h extraction provided better definition of the low to moderately contaminated locations by picking up small differences in anthropogenic metal concentrations. For the purposes of inter regional comparison, we recommend a 4 h 1 M HCl acid extraction as a standard method for assessing metal contamination in Antarctica. PMID- 15350412 TI - Assessment of degradation of 18 antibiotics in the Closed Bottle Test. AB - Large quantities of antibiotics are used in health care. After administration, they are discharged into the effluent and reach sewage treatment plants (STPs); if they are not degraded, they will eventually enter the environment. Antibiotics can affect bacteria in the environment and thus disturb natural elemental cycles. For this reason, it is necessary to take a closer look at the fate and effects of these substances in the environment. The biodegradability of 18 clinically important antibiotics and their effects on environmental bacteria was studied using the Closed Bottle Test (CBT) (OECD 301 D 1992). In addition, a toxicity control was performed in the CBT and the colony forming units (CFUs) were monitored. Disappearance of some of the 18 antibiotics was monitored by HPLC (high performance liquid chromatography) analysis. The antibiotics were used in two concentrations: (a) according to OECD 301 D in the mg/l-range and (b) on the basis of calculated concentrations in the influent of STPs in the microg/l-range. None of the 18 antibiotics were readily biodegradable. The HPLC analysis showed that some substances were partially or even completely disappeared by a non biotic mechanism. In the case of some antibiotics, partial biological removal took place in test vessels containing readily biodegradable sodium acetate and the test compound. However, in the toxicity control, toxicity had not been eliminated. PMID- 15350413 TI - Monitoring and evaluation of the environmental dissipation of the marine antifoulant 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT) in a Danish Harbor. AB - The concentration of marine antifoulant 4,5-dichloro-2-n-octyl-4-isothiazolin-3 one (DCOIT; the active ingredient in Sea-Nine 211 Antifouling Agent) leaching into a Danish Harbor from two painted ships was quantitated at varying distances from the ships. Sediment and suspended particulate matter were also analyzed for DCOIT. Water samples were concentrated on-site using C-18 solid phase extraction and subsequently analyzed by gas chromatography-tandem mass spectrometry. A strong decline in DCOIT water concentration as a function of distance from the ships was observed. The highest concentration (maximum 283 ng/l) was measured in the immediate vicinity of the ships and the concentration declined rapidly to less than the limit of detection (5 ng/l) at 400 m from the ships' surfaces. The measured decline curve was compared to that calculated using a one-dimensional model (ECoS). The comparison indicates that the primary mechanism of dissipation of DCOIT is not dilution resulting from dispersion but degradation with a rate constant in the order of 1 h(-1). Thus the field results correlate with the earlier microcosm studies demonstrating that DCOIT biodegrades rapidly in a marine environment. PMID- 15350414 TI - Observations on solid phase micro-extraction for MTBE analysis. AB - In a recent study of aqueous phase methyl tertiary butyl ether (MTBE) adsorption onto hydrophobic molecular sieves, the solid phase micro-extraction gas chromatography analytical technique was used to measure MTBE concentrations in water. The method was especially beneficial in measuring MTBE concentrations in the microg/l range, but anomalies were observed that investigators should be aware of before employing this technique. Specifically, it was observed that the calibration of the extraction fiber with known MTBE concentrations was non-linear over all concentration ranges. The technique was not suited to higher concentrations, and dilutions were necessary to increase the working range of the technique. Lastly, the fiber was observed to extract increasingly less MTBE from known standard solutions over time, requiring repeated calibrations to obtain reliable concentrations of unknown samples. PMID- 15350415 TI - An automated spectrophotometric system for monitoring buffer capacity in anaerobic digestion processes. AB - Anaerobic digestion is a suitable method for the treatment of wastewater and organic wastes, yielding biogas as a useful by-product. A common way of preventing instability problems and avoiding acidification in anaerobic digesters is to keep the organic load to the digester far below its maximum capacity. An improved way of operating digesters would be to use monitoring and control systems for increased organic load under controlled conditions such that the digester performance is improved. The partial alkalinity, which indicates the bicarbonate concentration, has in many cases been found to be a suitable parameter to monitor. Here, an automated monitoring system for alkalinity measurements is described. It is shown to be applicable for measuring a wide range of bicarbonate concentrations. The system shows potential for monitoring anaerobic digesters as it responds to the alkalinity of digester effluent, as well as being stable over a relatively long time span with few maintenance requirements. PMID- 15350416 TI - Toxicity of metals and metal mixtures: analysis of concentration and time dependence for zinc and copper. AB - The toxic effects of heavy metals, zinc and copper, in unary and binary solutions were studied using the Microtox acute toxicity test which relies upon the attenuation of light intensity emitted by Vibrio fischeri. The toxic effect Gamma (ratio of the light intensity lost at time t to the light intensity remaining at time t) of zinc could be related to its concentration [X] by a two-parameter equation Gamma=a(1-exp(-b[X])), where parameter a was a function of time and b equal to 0.88L/mg. The toxic effect of zinc asymptotically approached a maximum with respect of to concentration at all times. The toxic effect of copper was fundamentally different from that of zinc, and increased exponentially with concentration without any limiting maximum value. It could also be described by a two-parameter equation, however, the equation had the form Gamma=aexp(b[X]), where parameter a was a constant and b a function of time. The different functional dependencies (of the toxic effect on the metal concentration) of zinc and copper indicate that different toxicity/inhibition mechanisms were possibly responsible for the attenuation of light intensity for the two metals. The toxic effects of binary mixtures were substantially higher than those expected on the basis of additivity of individual metals. No simple correlations were obtained that could relate the toxic effect of binary mixture to those of individual metals. A better understanding of metal-microbe interactions is needed for achieving predictive capability for toxic effect of mixtures. PMID- 15350417 TI - Flow pattern visualization of a simulated digester. AB - Mixing patterns inside a simulated flat bottom digester were imaged using the non invasive techniques of computer automated radioactive particle tracking (CARPT) and computed tomography (CT). Mixing/agitation was provided using gas (air) recirculation at three different flow rates (Q(g)) of 28.32, 56.64 and 84.96 l/h, corresponding to superficial gas velocities of 0.025, 0.05 and 0.075 cm/s, respectively. Better mixing was observed in the upper zone near the top of the draft tube. However, at the bottom of the digester there was a total stagnancy at all the three gas flow rates. The maximum value of the time-averaged axial velocity inside the draft tube, at a gas flow rate of 84.96 l/h, was observed as 34.4 cm/s. The turbulent kinetic energy was observed to be maximum (724 dyn/cm(2)) inside the draft tube, and decreases radially toward the wall of the digester. The present study showed that the CARPT and CT techniques could be successfully used to identify the flow pattern in the digester and to calculate velocity and turbulence parameters quantitatively. On the other hand, the increase in gas circulation rate from 28.32 to 84.96 l/h did not significantly reduce the dead zones inside the flat bottom digester. To achieve the desired mixing and reactor performance, the operating conditions and reactor configuration need to be optimized. PMID- 15350418 TI - Influence of growth history on sloughing and erosion from biofilms. AB - The development of biofilms is determined by the balance of growth and detachment. But while the growth of biofilms is well studied, the influence of growth history and detachment on biofilm development is not. Here we report on laboratory scale experiments where heterotrophic biofilms were grown in a tubular reactor. Biofilm detachment was categorized based on particle size as erosion or sloughing. Erosion results in small particles and was approximated by the effluent suspended solids while sloughing was determined from the larger pieces of biomass that settled in a mixing tank. It was found that for all experiments, overall detachment was a combination of erosion and sloughing where erosion had a slightly larger contribution to the overall solids removal. However, sloughing had a significant influence on the biofilm morphology. Once the smooth biofilm surface was disturbed by a sloughing event (e.g., initiated through increasing liquid shear in the reactor), the biofilm became unstable resulting in spontaneous sloughing during subsequent operation. We propose that experimental investigations should consider sloughing events as an integral part of biofilm development rather than a disturbance. PMID- 15350419 TI - Influence of dissolved organic carbon content on modelling natural organic matter acid-base properties. AB - Natural organic matter (NOM) behaviour towards proton is an important parameter to understand NOM fate in the environment. Moreover, it is necessary to determine NOM acid-base properties before investigating trace metals complexation by natural organic matter. This work focuses on the possibility to determine these acid-base properties by accurate and simple titrations, even at low organic matter concentrations. So, the experiments were conducted on concentrated and diluted solutions of extracted humic and fulvic acid from Laurentian River, on concentrated and diluted model solutions of well-known simple molecules (acetic and phenolic acids), and on natural samples from the Seine river (France) which are not pre-concentrated. Titration experiments were modelled by a 6 acidic-sites discrete model, except for the model solutions. The modelling software used, called PROSECE (Programme d'Optimisation et de SpEciation Chimique dans l'Environnement), has been developed in our laboratory, is based on the mass balance equilibrium resolution. The results obtained on extracted organic matter and model solutions point out a threshold value for a confident determination of the studied organic matter acid-base properties. They also show an aberrant decreasing carboxylic/phenolic ratio with increasing sample dilution. This shift is neither due to any conformational effect, since it is also observed on model solutions, nor to ionic strength variations which is controlled during all experiments. On the other hand, it could be the result of an electrode troubleshooting occurring at basic pH values, which effect is amplified at low total concentration of acidic sites. So, in our conditions, the limit for a correct modelling of NOM acid-base properties is defined as 0.04 meq of total analysed acidic sites concentration. As for the analysed natural samples, due to their high acidic sites content, it is possible to model their behaviour despite the low organic carbon concentration. PMID- 15350420 TI - Plasticizer metabolites in the environment. AB - Earlier work with pure cultures had shown that the interaction of microbes with plasticizers leads to the formation of metabolites including 2-ethylhexanoic acid and 2-ethylhexanol that resist further degradation. The presence of these metabolites is now reported in a variety of environmental samples. Thus, even in a complex ecosystem, when plasticizers are degraded, the breakdown is not complete and significant amounts of 2-ethylhexanoic acid and 2-ethylhexanol are observed. These compounds have been shown to exhibit acute toxicity using Microtox, Daphnia, rainbow trout and fathead minnow toxicity assays. Since it is already well established that plasticizers are ubiquitous in the environment, it is expected that their recalcitrant metabolites will also be ubiquitous. This is of concern because, while the plasticizers do not exhibit acute toxicity, their metabolites do. PMID- 15350421 TI - Magnetic zeolites: a new adsorbent for removal of metallic contaminants from water. AB - In this work the adsorption features of Na Y zeolite with the magnetic properties of iron oxides have been combined in a composite to produce a magnetic adsorbent. These magnetic composites can be used as an adsorbent for metallic contaminants in water and subsequently removed from the medium by a simple magnetic process. The zeolites:iron oxide magnetic composites, were prepared by using Na Y with weight ratio of 3:1 and were characterized by powder X-ray diffraction (XRD), magnetization measurements, chemical analyses, N(2) adsorption isotherms and Mossbauer spectroscopy. Nitrogen adsorption isotherms showed that the surface area decreased from 505 m(2)g(-1) for the pure Na Y to 353 m(2)g(-1) for the Na Y:Fe oxide 3:1 composite. The adsorption isotherms of metal ions Cr(3+), Cu(2+) and Zn(2+) from aqueous solution onto the composites also showed that the presence of iron oxide does not affect the adsorption capacity. PMID- 15350422 TI - Amplification and attenuation of tetracycline resistance in soil bacteria: aquifer column experiments. AB - A growing inefficacy of antimicrobial agents to treat infectious diseases has stimulated research on the development of antibiotic resistance in bacteria in the environment. Sustained exposure of soil microorganisms to tetracycline (TC) in flow-through columns (50mg/L influent) significantly decreased the effluent concentration of total heterotrophs and selected for TC-resistant (Tet(r)) soil bacteria. This suggests that TC released to the environment from animal farms may contribute to the development and amplification of TC resistance, with soil bacteria serving as reservoirs for antibiotic resistance continuance. Burkholderia cepacia, with genetic determinants for efflux pumps that facilitate TC excretion, was the only bacterium that grew on TC-amended R2A plates. Following 300 days of exposure, TC was removed from the influent to study the recovery pattern of the microbial community. The percentage of Tet(r) hererotrophs decreased from 25% to close to the control level of 1% within 1 month of discontinuing TC exposure. This was due both to a significant rebound in the total heterotrophic population and to a significant decrease in the concentration of Tet(r) bacteria. Thus, discontinuing TC exposure or curtailing its use should enhance natural attenuation mechanisms that mitigate the spread of resistance vectors. PMID- 15350423 TI - GC-MS comparison of the behavior of chlorine and sodium hypochlorite towards organic compounds dissolved in water. AB - The reaction pathways of aqueous chlorination of anisole, ethylbenzene and o methoxybenzoic acid being structural fragments of humic matter were studied using gas chromatography-mass spectrometry. Chlorine and sodium hypochlorite were used as commonly employed disinfecting agents. Comparison of the chlorinating agents was performed in terms of the assortment and relative amounts of reaction products. Detailed schemes of the selected substrates transformation were proposed. Quantum chemical calculations were applied to propose structures of the reacting particles and a numerical parameter to estimate an extent of conversion of aromatic substrates during chlorination. PMID- 15350424 TI - Nitrate removal characteristics of high performance fluidized-bed biofilm reactors. AB - Two laboratory-scale high performance fluidized bed biofilm reactors (FBBR) with sand as the biofilm carrier were used to investigate the denitrification of high strength nitrate wastewater with specific emphasis on the effect the nitrogen loading rate and the superficial velocity (V(s)). The results demonstrated that the FBBR system is capable of efficiently handling an exceptionally high nitrate nitrogen concentration of 1000 mg N/L. At a loading rate of 6.3 kg-N/m(3)(bed).d almost complete denitrification was achieved with a removal efficiency of 99.8% and an effluent concentration of 2mg N/L at V(s) values of 45, 55 and 65 m/h. The maximum efficient loading rate (R(max)) at which the US drinking water nitrate nitrogen standard concentration of 10mg N/L would be exceeded was found to be a function of the applied V(s). The R(max) was found to be 12 kg-N/m(3)(bed).d at a V(s) value of 45 m/h. As V(s) was increased to 55 and 65 m/h, the optimum R(max) dropped to 9.5 and 8 kg-N/m(3)(bed).d, respectively. Higher denitrification rates were achieved at relatively lower V(s). However, there is a minimum practical velocity below which agglomeration of biomass would occur. The suspended solids concentration in the effluent was below 30 mg/L throughout the study. PMID- 15350425 TI - Chlorination effect on the fluorescence of nucleic acid staining dyes. AB - An alternative to culture methods for the control of drinking water disinfection would use fluorescent dyes that could evidence the nucleic acid damages provoked by sodium hypochlorite treatment. The two dyes selected in this study, SYBR Green II RNA gel stain and TOTO-1 iodide, efficiently stain nucleic acids (DNA and RNA) and quite poorly the other biomolecules considered (Bovine serum albumin, palmitic acid and dextrane). After treatment of nucleic acid solutions with increasing amounts of sodium hypochlorite, a decrease of fluorescence intensity is observed for both DNA and RNA stained with either SYBR-II or TOTO-1. However, the two fluorochromes do not lead to the same results, which shows that the two dyes are not bound to nucleic acids in the same way. Contrary to TOTO-1, SYBR-II reveals to be sufficiently sensitive to indicate both DNA or RNA damages as soon as the latter are in contact with hypochlorite even at concentrations of HClO lower than 10 micromol/L. Moreover, SYBR-II offers the opportunity to make quantitative titration of chlorine treated DNA and therefore seems to be the appropriate candidate to control the efficiency of the hypochlorite disinfection process of drinking water samples. PMID- 15350426 TI - Impact of a magnetic ion exchange resin on ozone demand and bromate formation during drinking water treatment. AB - The objective of this research was to examine the impact of a magnetic ion exchange resin (MIEX) on ozone demand and bromate formation in two different ozonated waters at bench scale. The first raw water had a high bromide ion concentration, a high ozone demand, and was highly colored. Based on experimental findings from the first water, the second water was selected as a model water in which more controlled experiments were performed. The waters were treated with the MIEX resin using jar test procedures to find the optimal MIEX dosage based upon the removal of ultraviolet (UV)-absorbing substances, dissolved organic carbon (DOC), and bromide. The optimal resin dosage was chosen for bulk MIEX treatment and subsequent ozonation in a semi-batch reactor. The ozone demand and formation of bromate were analyzed as a function of ozone dosage and dissolved ozone concentration for the MIEX pre-treated water, and compared to the results obtained by ozonating the water without MIEX pre-treatment. The results indicate that pre-treatment of the water with the MIEX resin significantly reduces total organic carbon, DOC, UV absorbance, color, and to some extent, bromide. MIEX pre treatment of the water prior to ozonation substantially lowered the ozone demand and formation of bromate during subsequent ozonation. PMID- 15350427 TI - Degradation of sodium dodecylbenzene sulfonate in water by ultrasonic irradiation. AB - The potential of using ultrasonic irradiation for the removal of sodium dodecylbenzene sulfonate (SDBS) from aqueous solutions has been investigated. Experiments were performed at initial concentrations of 15, 30 and 100 mgl(-1), ultrasonic frequencies of 20 and 80 kHz, applied power values of 45, 75 and 150 W and liquid bulk temperatures of 20, 40 and 60 degrees C. At the conditions in question, SDBS conversion was found to decrease with increasing temperature and initial solute concentration and decreasing power and frequency. Investigations using the radical scavengers 1-butanol and KBr revealed that SDBS degradation proceeds through radical reactions occurring predominately at the bubble-liquid interface and, to a lesser extent, in the liquid bulk. Addition of NaCl or H(2)O(2) had little or even an adverse effect on SDBS conversion. Conversely, addition of Fe(2+) either alone or in conjunction with H(2)O(2) (Fenton reagent) had a positive effect on degradation. Finally, shake flask tests with activated sludge were performed to assess the aerobic biodegradability before and after sonochemical treatment. At the conditions under consideration, the use of ultrasound enhanced the aerobic degradability of the substrate in question. PMID- 15350428 TI - Effect of nitrite on phosphate uptake by phosphate accumulating organisms. AB - In biological nitrogen removal processes, nitrite can be formed and accumulated through both nitrification and denitrification. Despite the fact that, in practice, biological phosphate removal (BPR) is often combined with biological nitrogen removal, there are only a few publications reporting the effect of nitrite on BPR. In this study, phosphate-accumulating organisms (PAO) were cultivated in an anaerobic-anoxic-aerobic sequencing batch reactor (SBR). The effect of nitrite on the enrichment of the sludge with PAO, the phosphate uptake rates and the sludge respiration was investigated. The results indicate that (1) presence of nitrite inhibits both aerobic and anoxic (denitrifying) phosphate uptake, (2) aerobic phosphate uptake was more affected than anoxic phosphate uptake, (3) presence of nitrite could be one of the factors enhancing the presence of glycogen accumulating organisms (GAO)--competitors to PAO for substrate in the anaerobic phase, and (4) it is required to monitor and control nitrite accumulation in a full-scale wastewater treatment plants. PMID- 15350429 TI - Microbiology, chemistry and biofilm development in a pilot drinking water distribution system with copper and plastic pipes. AB - We studied the changes in water quality and formation of biofilms occurring in a pilot-scale water distribution system with two generally used pipe materials: copper and plastic (polyethylene, PE). The formation of biofilms with time was analysed as the number of total bacteria, heterotrophic plate counts and the concentration of ATP in biofilms. At the end of the experiment (after 308 days), microbial community structure, viable biomass and gram-negative bacterial biomass were analysed via lipid biomarkers (phospholipid fatty acids and lipopolysaccharide 3-hydroxy fatty acids), and the numbers of virus-like particles and total bacteria were enumerated by SYBR Green I staining. The formation of biofilm was slower in copper pipes than in the PE pipes, but after 200 days there was no difference in microbial numbers between the pipe materials. Copper ion led to lower microbial numbers in water during the first 200 days, but thereafter there were no differences between the two pipe materials. The number of virus-like particles was lower in biofilms and in outlet water from the copper pipes than PE pipes. Pipe material influenced also the microbial and gram negative bacterial community structure in biofilms and water. PMID- 15350430 TI - Removal of arsenic using hardened paste of Portland cement: batch adsorption and column study. AB - Hardened paste of Portland cement (HPPC) has been used as a low-cost adsorbent for the removal of arsenic from water environment. Results from the batch experiments, conducted at an initial concentration of 0.2 ppm of arsenate, suggest arsenate removal up to 95%. Kinetic profiles were developed for various conditions. Effects of adsorbent dose, common ions such as Ca(2+), Mg(2+), Fe(3+), Fe(2+), Cl(-), SO(4)(2-), NO(3)(-), PO(4)(3-) and of pH were studied in detail. Adsorption isotherm studies revealed that the Freundlich isotherm was followed with a better correlation than the Langmuir isotherm. Arsenite could also be removed up to approximately 88% using the same material, HPPC. Finally, column studies were undertaken involving the new HPPC to check the suitability of the material for the removal of total arsenic content from water body. Kinetic experiments for the removal of arsenic by column studies revealed a film diffusion mechanism. PMID- 15350431 TI - Application of fluorescence spectroscopy in the studies of natural organic matter fractions reactivity with chlorine dioxide and ozone. AB - The composition of natural organic matter (NOM) fractions before and after the reaction with chlorine dioxide as well as ozone was studied by means of total luminescence spectroscopy (TLS) and synchronous scanning fluorescence measurements. The excitation-emission matrices spectra (EEMs) of natural as well as oxidised NOM fractions revealed two well-resolved bands with maxima at Ex/Em of 250-265/422-452 nm and at Ex/Em 300-336/414-446 nm ascribed to humic and fulvic material. The study of emission and synchronous spectra also confirmed the presence of protein-like constituents in all examined NOM fractions. The study of EEMs proved, that oxidation of all NOM fractions with ClO(2) caused mainly the break-up of molecules into smaller fragments and a decrease of the aromaticity. Changes in EEMs after the oxidation of individual NOM fractions with O(3) confirmed the formation of a significant amount of ozonation by-products, i.e. carboxylic acids, aldehydes and ketones during the oxidation process. In addition, the fluorescence studies confirmed relatively high reactivity of all NOM fractions with ClO(2) as well as with ozone. PMID- 15350432 TI - An evaluation of solid phase microextraction for analysis of odorant emissions from stored biosolids cake. AB - Odors are a common occurrence at wastewater treatment plants, biosolids processing facilities and biosolids recycling locations. Accurate, objective measurement techniques are needed to monitor emissions, to develop new waste handling procedures and to reduce the production of the volatile gases. The objective of this study was to evaluate the use of solid phase microextraction for measuring common odorants that are found in biosolids facilities. The odorants were collected and concentrated by solid phase microextraction (SPME) and then quantified by gas chromatography with detection by mass spectrometry. A 75-microm Carboxen-Polydimethylsiloxane coating was used for the analysis of trimethylamine, dimethyl sulfide, dimethyl disulfide and methyl mercaptan. Gaseous standards were generated for individual compounds and for dry and wet mixture from permeation apparatus. The differences in sensitivity between fibers, the competition between analytes and water vapor for the active sites on the fiber and the lack of production of artifacts make SPME suited for qualitative analysis and enables quick screening for the identification of compounds with adverse organoleptic characteristics. PMID- 15350433 TI - A new approach to evaluating the toxicity and genotoxicity of disinfected drinking water. AB - The aim of this study was to evaluate the formation of toxic and genotoxic compounds in surface drinking waters treated with two widely used disinfectants, sodium hypochlorite (NaClO) and chlorine dioxide (ClO(2)), and a new disinfectant, peracetic acid (PAA). For this purpose a pilot plant was set up to add these biocides continuously to pre-filtered lake water flowing into three different basins. During three seasonal experiments, short-term in vivo tests (with plant, fish and molluscs) and in vitro tests (with bacteria, yeast and human cells) were carried out to evaluate the formation of genotoxic disinfection by-products (DBPs). Gas chromatography/mass spectrometry (GC/MS) was used to identify DBPs produced during the different treatments, microbiological analyses were performed to test the biocidal activity of the disinfectants, and chemical analyses were carried out to evaluate the quality of the water. The pilot drinking water plant under study was useful in studying the toxicity and genotoxicity of disinfected drinking water with this combined chemical/biotoxicological approach. This paper describes the setting up of the pilot plant and sets out/reports the results of the microbiological and chemical analyses. PMID- 15350435 TI - New approaches to replacing failing organs. AB - The foremost cause of death and disability is organ failure. Failure of the heart, lungs, kidney, and liver is typically addressed by organ transplantation; however, the number of human organs available for this purpose is quite limited. For this reason, some new technologies are being advanced for the treatment of organ failure. This communication summarizes potential limitations of these technologies. PMID- 15350436 TI - Can a living kidney donor become a kidney recipient? AB - The living kidney donor represents a good resource for kidney transplantation. These grafts display better function and long-term graft survival at 5 and 10 years of follow-up. Furthermore, living donors prefer the possibility to increase kidney donation for a large waiting list of patients with end-stage renal disease (ESRD). However, kidney donation is a major surgical procedure associated with benefits and risks. The risks of donation have been studied in large series of living donors to focus on morbidity and mortality rates associated with the surgical procedure. New surgical laparoscopic techniques promote living kidney donation. While the benefits to the recipient are obvious, those for the donor are subjective and not quantifiable. However, donors describe donation as a great experience in life. The risk of kidney donation may be divided into the perioperative and the long-term risks. The evaluate the long-term risks for kidney donors requires a long follow-up. The main source of kidney donors in our transplant center has been living-related and -unrelated donors, with a minor percentage of cadaveric donors. In this report we present four kidney donors who developed ESRD thereafter, three becoming kidney recipients. PMID- 15350437 TI - Organs transplanted from intoxicated donors. AB - PURPOSE: The purpose of our study was to evaluate short- and long-term results of transplants from cadaver donors who have died of poisoning by various substances. MATERIALS AND METHODS: The actuarial survival rate of organs from intoxicated donors was calculated using the Kaplan-Meier method. RESULTS: Among the 507 donors between January 1998 and December 2002, 5 (0.98%) had a cause of brain death of poisoning, namely, organo-phosphates (n = 2), methanol (n = 1), cyanide (n = 1) and acetylsalicilic acid(n = 1), from whom were procured 10 kidneys, 1 liver, 2 corneas, and 1 set of bones. The follow up for patients receiving solid organs was 15.2 months (range, 0-48 months). At 3 months, 90% of kidneys had normal function. No delayed graft function rejection episodes or major complications were reported in any recipient. None showed evidence of acute or chronic poisoning. Two died, 1 early mortality was due to anesthetic complications and the other at 17 months to an unknown cause. Actuarial kidney survival rates were 90% and 80% at 12 and 24 months, respectively. The liver recipient was well at the end of follow up. CONCLUSION: Using organs of poisoned donors is feasible with comparable graft survival rates to other recipient. PMID- 15350438 TI - Evaluation and quality control of organ transplant coordination services, transplant procurement management based on the new vital cycle. PMID- 15350439 TI - Brain death: epidemiology and quality control of solid organ donor generation. AB - Successful solid organ transplant programs need effective organ procurement systems. Evaluating and maintaining the quality of an organ procurement system requires multilevel institutional coordination as well as reliable information regarding potential donors. The objective of this study was to analyze the potential to generate adult solid organ donors at a countrywide level as well as at individual hospital levels in Uruguay from 2000 to 2002. The analyzed factors included: encephalic deaths (brain deaths) officially registered by the National Bank of Organs and Tissues (BNOT), the distribution of these individuals among the intensive care units (ICUs) located at various hospitals, the number of Real Donors at each hospital, and the relation between the brain deaths and the number of ICU beds equipped with artificial respirators. The Uruguayan solid organ donation rate is relatively high among Latin American Countries, but low in relation to developed countries. The conversion of potential donors to actual donors is similar to that seen in developed countries. However, the registered incidence of brain deaths in relation to the number of ICU beds is surprisingly below the recognized standard figure. To approach the reasons for this difference, efficient quality control mechanisms are needed in the various ICUs as well as comprehensive medical case history reviews of all patient records. PMID- 15350440 TI - Stable renal transplant recipients can be safely converted from MMF to enteric coated mycophenolate sodium tablets: Interim results of a multicenter Latin American study. AB - Enteric-coated mycophenolate sodium (EC-MPS) is designed to reduce mycophenolate acid (MPA)-related upper gastrointestinal (GI) adverse events (AEs). A multicenter, open-label, Latin American study in stable renal transplant patients is ongoing to assess the safety of the conversion from mycophenolate mofetil (MMF) to EC-MPS. An interim analysis was performed when 93 patients had completed 3 months. Prior to conversion, they had received MMF at a dose of 2 g/d, with the exception of eight adult patients who were receiving an average daily dose of 1.25 g. All adult patients were converted to EC-MPS (1.44 g/d; 0.450 g/m(2) bid for children). After conversion, the reported total incidence of AEs was 40.9%, including 28% infections, 1.1% hematologic, 19.4% GI, including 10.8% upper-GI AE (all mild) and 5.4% diarrhea. No patient discontinued the study medication due to adverse events. Only six patients (6%) required a dose adjustment. There were no episodes of acute rejection, death, or graft loss. During the period of analysis, the conversion from MMF to EC-MPS was safe, the enteric-coated tablet formulation prevented release of MPA in the upper GI tract, and only one patient had to reduce the dose due to an upper GI AE, concomitant with diarrhea. EC-MPS offers transplant physicians and their patients an alternative MPA therapy that is as effective and safe as MMF, but in a formulation that may provide GI tolerability benefits. PMID- 15350441 TI - Nutrition for the post-renal transplant recipients. AB - An adequate nutritional status may improve outcomes after renal transplantation. This review article presents the impact of major nutritional risk factors, such as malnutrition, obesity, dislipidemia, and other associated or independent metabolic complications on the results of renal transplantation. Malnutrition that exists prior to transplant may be associated with an increased risk of infection, delayed wound healing, and muscle weakness. Obesity, which may also be preexisting or developing after transplantation, can lead to adverse effects, such as poor wound healing or increased risk of cardiovascular disease, although some recent studies suggest no major consequences. Hypercholesterolemia and hypertrigliceridemia (risk factors for cardiovascular disease) are common post renal transplantation complications. All of these possible complications may be prevented or treated through early nutritional intervention and follow-up. This article reviews important nutritional recommendations to provide guidelines for adult posttransplant recipients. PMID- 15350442 TI - Renal transplants with delayed graft function show decreased renal function despite monitoring with postabsorptive levels. AB - Cyclosporine (CyA) monitoring with postabsorptive levels can predict the risk of an acute rejection episode (ARE). Large doses of CyA are needed to obtain adequate drug exposure. The impact of this strategy on renal function, especially in patients with delayed graft function (DGF), is unknown. We report our experience comparing C3 (3-hour postdose) monitoring with a historical series of cadaveric renal transplants. Sixty-three consecutive patients who received cadaveric renal transplants were followed for 1 year. Group A (historical n = 31) patients received 6 mg/kg/d CyA with the dose adjusted according to the trough level (target, 250-350 ng/mL), group B (study n = 32) received 10 mg/kg/d CyA with dose adjustments based upon C3 (target, 1100-1500 ng/mL). All patients received cyclosporine prednisone and a third agents. The general characteristics of the donors and recipients were comparable. The incidence of biopsy-proven ARE at 1 year in group A was 42% and 19% in group B (P <.05). Patients achieving C3 levels >1000 ng/mL at 1 week displayed significantly lower ARE rates (8% vs 50%; P <.05). The rate of DGF was similar in both groups, but the duration was longer in group B (15 vs 21 days, P <.05). The serum creatinine (SCr) level was significantly higher in group B at 3 months (1.47 mg/dL group A vs 1.76 mg/dL group B; P <.05). Patients in group B with DGF showed significantly higher SCr values at 1 year (1.18mg% vs 2.03 mg%; P <.05). C3 level monitoring of CyA yields excellent results in terms of decreased ARE, but an increased SCR was observed among patients with DGF. PMID- 15350443 TI - Counting of lymphocyte subpopulations using standard versus low-dose antiCD3 monoclonal antibody treatment. AB - The monoclonal anti-CD3 antibody is used as part of prophylaxis and also in treatment of rejection. In the present article we analyzed changes in different lymphocyte subpopulations after anti-CD3 treatment. T lymphocytes were decreased under anti-CD3 antibody administration, with a simultaneous increase in B lymphocytes but no changes in natural killer (NK)cells. No differences were found between patients administered anti-CD3 antibody (Ab) at 5 versus 2.5 mg/d. It is uncertain whether these changes may be implicated in the lack of response or in the prophylactic effects of anti-CD3 Ab. PMID- 15350444 TI - Effect of rapamycin on cytokine profile in kidney transplant recipients treated with triple drug therapy. AB - The aim of this study was to explore differences in the cytokine profile among de novo kidney transplant recipients treated with either Rapamycin (Rapa) + cyclosporine (CsA) + prednisone (P) or CsA + azathioprine (Aza) + P. PATIENTS AND METHODS: Among the 13 adult kidney transplant recipients studied, seven received Rapa + CsA + P while the remaining six received CsA + Aza + P with their living donors serving as controls (n = 13). Spontaneous production of IL-2, IFNgamma, IL 10, and TGF-beta were measured by ELISA in supernatants from 24-hour cultured unstimulated peripheral blood mononuclear cell (PBMC) at time zero (the day before the transplant), and at 3 and 6 months posttransplant. Cytokines were also measured 1 month after CsA withdrawal in the Rapa + CsA + P group. RESULTS: From time zero to the end of the study, IL-2, IFNgamma, and IL-10 were present at low or undetectable levels in all three groups. TGF-beta tended to increase in supernatants from patients under Rapa + CsA + P at 6 months posttransplant and at 1 month after CsA withdrawal without correlation to Rapa blood levels. TGF-beta remained stable throughout the study period for patients included in the CsA + Aza + P group. There was no difference in this cytokine level between these study groups at any given time. CONCLUSIONS: This study showed no differences in the spontaneous cytokine profiles evaluated in patients treated with both therapeutic schemes. PMID- 15350445 TI - Liver transplantation in Latin America: current status. AB - The current status of liver transplantation in Latin America is reviewed. For an estimated population of 470 million people, approximately 1100 liver transplantations were performed in 2002, namely, 2.3 liver transplantations per million people per year (pmp). The top transplantation rates were in Argentina 4.5, Brazil 3.9, and Chile 3.6 pmp. In 2001, liver transplantations were not performed in 2 of 10 South American countries nor were liver transplantations performed in 5 of 6 Central American countries. In the Latin American countries performing this procedure, living donor liver transplantation has been performed to palliate the organ shortage for pediatric patients in most of these countries. However, only 3 countries have used the procedure for adult liver transplantation. The lack of adequate financial coverage, education, and organization are the main limiting factors to the development of liver transplantation in Latin America. PMID- 15350446 TI - Clinical characterization and survival of adult patients awaiting liver transplantation in Chile. AB - Liver transplantation has become widely used for patients with decompensated disease. Because of the shortage of donors, each year more patients die on the waiting list. Our aim was to characterize and evaluate the final outcomes of all listed candidates for liver transplantation during a 34-month period. We retrospectively evaluated all adults listed between January 2000 and November 2002. Sixty-three patients (37 women, mean age 45.8 years) were listed: 48 due to chronic liver disease and 15 for a highly urgent transplantation due to acute liver failure. The main etiology of chronic disease was alcoholic (22%) or primary biliary cirrhosis (17%). Of 52 chronic patients, 26 (50%) were transplanted with a mean waiting time of 168 days. Among the others, 8 died (15%) while awaiting transplantation, 3 (5%) were removed from the list, and 15 patients still await transplantation (28%). Among acute liver failure patients, the main etiologies were autoimmune (25%) and medication induced (25%). Of 15 acute patients, 6 (37.5%) have been transplanted at a mean waiting time of 6.8 days with 100% survival posttransplantation. In this cohort, 6 patients (37.5%) died while awaiting liver transplantation, and 4 (25%) survived with medical support. In conclusion, the severity of liver disease and death rate among our waiting list was similar to that observed in developed countries. It seems reasonable to review our current allocation system based on waiting time on the list. We will have to decide whether to transplant sicker patients or those with hepatocarcinoma (as in the United States recently with the MELD system), thereby possibly decreasing the mortality rate on the waiting list at the expense of higher costs and more difficult postoperative care or to just keep our current policy. PMID- 15350447 TI - Predictive models in cirrhosis: correlation with the final results and costs of liver transplantation in Chile. AB - Medical scores for predicting survival are essential to stratify patients with end-stage liver disease (ESLD) for prioritization for liver transplantation (OLT). Recently the UNOS has adopted the Mayo Model for End-stage Liver Disease (MELD) score as the basis for liver allocation in the United States. We retrospectively evaluated and assessed the prognostic impact, the length of stay (LOS), and hospital charges for OLT using two severity scores (Child-Turcotte Pugh [CTP] versus MELD) to stratify cirrhotic patients before OLT. Twenty-six consecutive adult cirrhotic patients (11 women, mean age 46 years) underwent LT between 2000 and 2002. The main causes for transplantation were alcohol and primary biliary cirrhosis. The mean CTP and MELD scores at the moment of listing for OLT were 8.9 and 16.3 points, respectively. The best discriminative values with prognostic impact in terms of outcome and costs of OLT were a Child Pugh score >/=11 points or a MELD score >/=20 points. Patients in these strata showed a significant increase in LOS in the hospital (from a mean of 12 to 22 days) and intensive care stay (from a mean of 4 to 14 days) post-OLT when compared with patients with a lower CTP or MELD score (P <.05). There was also a trend toward higher hospital charges (P =.06). Organ allocation by MELD score will probably adversely affect the LOS and hospital charges of patients being transplanted due to ESLD. PMID- 15350448 TI - A decade of adult liver transplantation in a single center in Chile. AB - INTRODUCTION: Since the early days, liver transplantation (OLT) has conquered several barriers worldwide to become a proven therapy. We assessed the evolution of our adult liver transplant program. METHODS: We studied all adult patients who underwent OLT since the inception from November 1993 through May 2003. Donor data, recipient pretransplantation evaluation, surgical technique, results, and costs were examined over our evolution, stratifying 3 groups over time, based on the number of adult OLT per year. RESULTS: Between November 1993 and May 2003, 70 OLT were performed in 64 patients older than 15 years of age. Preoperative Child score, preoperative creatinine level, donor and recipient age, and proportion of emergencies were similar in the 3 groups. Over time, the predominant surgical technique was the piggyback technique (97% of OLT) with a decrease in the use of bypass from 63% to 5% during the last time period. Over the 10 years of our program's existence, warm ischemia time has been reduced to less than 1 hour, whereas cold ischemia time has remained constant at around 5 hours. Biliary and vascular complications decreased over time to around 10%. The mean length of hospital stay (LOS) decreased to 12 days (excluding emergencies). Since inception, our 1-year patient survival rate average is 91%; however, in just the last 3 years of our program (2000 through 2003), the 1-year patient survival rate is 97%. CONCLUSIONS: In summary, our surgical technique has evolved toward piggyback use without veno-venous bypass with a significant decrease in warm ischemia times. As expected, our results have improved over time and our LOS and costs have decreased. Finally, our current results are similar to the best ones reported in the medical literature today. PMID- 15350449 TI - Rapid steroid taper and neoral monotherapy in liver transplantation in Chile: a step in the right direction? AB - Diabetes, hypercholesterolemia, hypertension, obesity, osteopenia, and increased risk of viral recurrence are among the complications associated with posttransplant steroid use. Steroid withdrawal or rapid taper has been reported to be safe. The aim of this study was to compare the rejection incidence and severity among patients treated with two different steroid taper strategies. METHODS: This retrospective study included all the adult liver transplant recipients since the program's inception from 1993 to January 2002. The minimum follow-up was 1 year. Exclusions included patients receiving an immunosupressive regimen other than mycophenolate mofetil, steroids, and Neoral, or suffering an autoimmune etiology, or displaying patient or graft survival less than 1 year. The incidence and severity of rejection episodes were compared between the two groups of steroid taper protocols: group A received methylprednisolone (1 g) intraoperatively with a slow taper to 10 mg prednisone per day at 1 year. Group B received methylprednisolone (2 g) intraoperatively followed by a rapid reduction with intention to withdraw by month 4, continuing on Neoral monotherapy. Rejection diagnosis was made on histological bases. RESULTS: One-month and 1-year rejection rates were 47% and 53%, respectively, among the rapid taper group with Neoral monotherapy, which was similar to 60% and 64%, respectively, in the slow taper group. Rejection severity was also comparable between the two groups. CONCLUSIONS: Patients treated with a rapid steroid taper protocol followed by Neoral monotherapy or a slow taper protocol showed similar acute rejection incidences and severities. Their survival rates were also comparable. Further study is necessary to evaluate the impact of rapid steroid taper to prevent the complications of steroid use. PMID- 15350450 TI - Liver transplantation in Medellin, Colombia: initial experience. AB - OBJECTIVE: The goal of this article is to present the experience of a new liver transplant team. MATERIALS AND METHODS: This review includes all patients who received a liver transplant between March 15, 2000 and March 15, 2003. RESULTS: We performed 87 transplantations on 84 patients; 39 were females and 45 were males of average age 43.6 years, including 6 children. The majority of the patients were from Colombia with time on the waiting list of less than 1 month. The average donor age was 26.7 years. The preservation solutions included Wisconsin, HTK-Brettschneider (Custodiol), and Corpaul (similar to Henn-Ross). In this study, 95.4% were whole livers, with 97.7% using the piggyback method. We placed 23 arterial grafts and 2 venous grafts for vascular reconstructions; 95.4% were duct-to-duct anastomosis (95.4%). Among the cohort, 8.3% experienced acute rejection and 1.2% experienced chronic rejection. Two patients required retransplantation due to hepatic artery thrombosis with biliary tree necrosis. CONCLUSIONS: We consider that we have passed the crisis of beginning a new program with a reduction in postoperative complications and improving patient and graft survival. At present, we are a center that performs liver transplantations in adults and children, with a good organ donation culture in our city that allows us to offer a waiting time on the list less than one month. Neither a veno venous bypass nor a T-tube were necessary for our cases. We also have developed a new, less expensive form of perfusing the liver in the donor. PMID- 15350451 TI - Transcystic cholangiogram access via rubber band with early withdrawal after liver transplantation: a safe technique. AB - OBJECTIVE: Since different techniques have been described for cholangiogram access after liver transplantation, we compared two different methods for patients with duct-to-duct biliary anastomoses. METHODS: Adult liver transplant patients from program inception in 1993 to May 2003 in whom a duct-to-duct biliary anastomosis with a T-tube choledochostomy were compared with those having a transcystic duct catheter using a rubber band. We excluded 10 patients in which a different technique was used or graft or patient survived less than 21 days. Group A (n = 28,) had a number 10 T-tube exteriorized through the recipient main bile duct; and group B (n = 33) a number 5 Bard ureteral stent tied to the cystic stump with reabsorbable suture and secured with a hemorrhoidal rubber ligature. RESULTS: The biliary complication rate was lower among the transcystic catheter group (9.1%, 3/33) compared to the T-tube group (35.7%, 10/28). Postcatheter withdrawal peritonitis was present in two patients in the T-tube group, one of whom required emergency laparotomy. A satisfactory postoperative cholangiogram was obtained in both groups. The transcystic catheter was withdrawn on average at 29 days, compared to 136 days in the T-tube group. CONCLUSIONS: Both techniques are equally effective in obtaining a satisfactory postoperative cholangiogram. However, the transcystic catheter technique allows a significantly earlier withdrawal with fewer complications compared to the T-tube technique. PMID- 15350452 TI - Trends in mechanical ventilation and immediate extubation after liver transplantation in a single center in Chile. AB - INTRODUCTION: Some groups have reported early extubation post-liver transplantation in patients with previously defined criteria, in an attempt to shorten the ICU stay and decrease costs. We review our experience with trends in mechanical ventilation and resource utilization. METHODS: We retrospectively reviewed the length of mechanical ventilation, ICU stay, hospital stay, transfusions, and costs of liver transplants performed since the program's inception in 1993 and 2002 including 82 OLT in 71 patients. We also report our experience with immediate postoperative extubation, which we have done from October to December of 2002. We compare different periods: the early days (1993 to 1997), namely, fewer than 10 OLT per year, with the subsequent years assessed individually. RESULTS: There has been a progressive decrease over time in the length of mechanical ventilation, ICU stay, hospital stay, and costs. Since the program's inception actuarial adult patient 1- and 5-year survival rates were 88.7% and 78%, respectively. The 1-year survival rate increased to 97% during the period of 2000 to 2002 (n = 30). From October to the present, we extubated four of seven adult patients who met criteria with none of them requiring reintubation. CONCLUSIONS: We demonstrate improved results, decreased length of mechanical ventilation, ICU, and hospital stay, and costs. The immediate postoperative extubation may be feasible for patients who meet previously defined criteria. PMID- 15350453 TI - Cardiovascular disease and/or elderly donors in renal transplantation: the outcome of grafts and patients. AB - Kidney transplant programs nowadays increasingly use elderly, hypertensive and cardiac disease donors (expanded criteria donors). The impact of these donors on patient and graft outcome was investigated in our transplant population. Among 257 consecutive cadaveric kidney transplants, 56 were from expanded criteria donors. The frequency of anuria, delayed graft function, and the days of renal failure were higher using organs from the expanded criteria donor group. Serum creatinine was higher in this group, although the statistical significance disappeared at 36 months. There were no significant differences in graft or patient survival during the first 3 years. The use of expanded criteria donors should not be discouraged, but recipient selection and immunosuppression use should be adapted and cold ischemia minimized. PMID- 15350454 TI - Lung transplant at the Favaloro Foundation: a 13-year experience. AB - BACKGROUND: The outcomes of lung transplantation (LT) are well known in developed countries, but not in Latin America. Our objective was to report the LT experience at a single center in Argentina. METHODS: From June 1994 to February 2003, the 54 LT that were performed included 36 single-lung transplants SLT (45.5%) and 18 double-lung transplants (DSLT) (22.7%). Indications for SLT (n = 36) were emphysema (n = 23) and pulmonary fibrosis (n = 13); for DSLT (n = 18), bronchiectasis (n = 8), cystic fibrosis (n = 8), pulmonary emphysema (n = 1), and bronchiolitis obliterans syndrome caused by graft-versus-host disease after bone marrow transplantation (n = 1). A univariate analysis, chi-square test with Yates' correction was used for qualitative variables; Wilcoxon-Mann-Whitney test, for quantitative and ordinal variables. Survivals were estimated by the Kaplan Meier method. RESULTS: In-hospital mortality (HM) was 21.1%. Among SLT, early death was due to sepsis in six patients and by ischemia-reperfusion injury (IRI) and acute rejection in other two. In DSLT, two patients died due to IRI and one, sepsis. The overall estimated survival rates at 1, 2, and 4 years were 70.1% +/- 6.5%, 54.3% +/- 7.2%, and 44.3% +/- 7.9%, respectively. The median overall survival was 26.5 (10 to 34) months. When HM was excluded, survival at 4 years was 51.3% +/- 8.7%. The estimated survival at 3 years was 43.3% +/- 9.3% for SLT and 58.7% +/- 13% for DSLT (P = 6). Survival differences according to the baseline diagnosis were not significant (P =.6). Median follow-up time (percentiles 25 to 75) was 16 (2 to 27) months. CONCLUSIONS: Our LT program shows similar results to those reported by the International Society for Heart and Lung Transplantation for developed countries. PMID- 15350455 TI - Orthotopic heart transplantation in elderly patients: a 10-year experience at a single center. AB - INTRODUCTION: A consensus has not yet been reached regarding the indications for orthotopic heart transplantation (OHT) in elderly patients or the age limit contraindicating the procedure. The objective of this study was to assess OHT outcomes to determine whether elderly patients benefit from the procedure. METHODS: From February 1993 to February 2003, 178 OHTs were performed on recipients of mean age 47.4 +/- 15 years (range, 4 to 74) including 80.3% men. The population was divided into two groups: group A included patients >/= 60 years, and group B those younger than that age. Survival was analyzed for the overall population and for both age groups during a 10-year follow-up period. RESULTS: Group A included 36 patients (20.8%) and group B 142 patients (79.2%). Mean age was 63.7 +/- 2.9 years (60 to 74) in A, and 43 +/- 13.9 years (4 to 59) in B. In-hospital mortality was significantly higher among group A (n = 11, 31.4%) compared to B (n = 17, 12.1%, P =.008). Survival at 1, 5, and 10 years was 61.5% +/- 8%, 58.1% +/- 8.3%, and 49.8% +/- 10.5% group A; and 84.2% +/- 3%, 73.7% +/- 4.1%, and 69.9% +/- 4.7 for group B. Elderly patients showed a lower survival rate (49.8%) compared with the younger group (69.9%) at 10-year follow up (P =.007). Conditional survival at 9 years failed to show significant differences (A 72.2% vs B 79.6%, P =.4). CONCLUSION: In our population, elderly recipients showed a higher in-hospital mortality. However, when the first post OHT year was excluded, we found similar survival rates for both age groups. PMID- 15350456 TI - Beta-cell replacement therapy (pancreas and islet transplantation) for treatment of diabetes mellitus: an integrated approach. PMID- 15350457 TI - Establishing an islet transplantation program in a developing country. AB - Diabetes is an emerging epidemic throughout the world. In our city alone, there are approximately 25,000 known diabetics (5% to 10% type 1) among a total population of 1.7 million inhabitants, and the incidence is increasing among all age groups. Islet transplantation is a potential treatment for type 1 diabetes mellitus. For this reason, we intended to establish an islet transplantation program. This required competent and well-trained professionals, a specially planned facility adhering to rigid regulations regarding safety and sterility, and a detailed study of the ethical laws and rules involving transplantation. In this article, we describe the process including any difficulties or barriers encountered due to limited resources in a developing country. We also describe all stages of personnel training and the necessary equipment and work area of a similar specialized center following the guidelines of the Brazilian National Agency for Health Care. Finally, we discuss our expectations for the initial phase of our islet transplantation program. PMID- 15350458 TI - Hematologic reconstitution following high-dose and supralethal chemoradiotherapy using stored, noncryopreserved autologous hematopoietic stem cells. AB - Although cryopreservation is the standard for autotransplantation, it has logistic and financial disadvantages in undeveloped countries such as Colombia. In 47 patients, peripheral blood was refrigerated at 4 degrees C up to 144 h before autotransplantation. For mobilization, 27 men and 20 women of median age 37 years affected with hematologic malignancies received G-CSF. The 17 patients in Group 1 showed pre-refrigeration CFU-GM of 2.62 x 10(5)/kg (range 0.36 to 16.6 x 10(5)/kg) and at re-infusion, 1.36 x 10(5)/kg (range 0 to 6.32 x 10(5)/kg) of 83% viability (range, 78% to 96%). These patients showed >0.5 x 10(9)/L granulocytes on day +11 (range, 9 to 15) and >20 x 10(9)/L platelets on day +16 (range, 11 to 44). The 25 patients in Group 2 showed CD34 of 3.9 x 10(6)/kg (range, 0.16 to 9 x 10(6)/kg) and mononuclear cell count (MNC) of 8.7 x 10(8)/kg, reaching >0.5 x 10(9)/L granulocytes at day +13 (range, 10 to 17) and >20 x 10(9)/L on day +15 (range, 14 to 20). Among the 5 patients in Group 3, the average of MNC of 12.7 x 10(8)/kg was reached and >0.5 x 10(9)/L granulocytes on day 11 (range, 10 to 16) and >20 x 10(9)/L on day 14 (range, 10 to 18). No differences were observed between the groups. Refrigeration of stem cells appears to be a simple, effective, and inexpensive method that should be considered for autotransplants within a few days of harvesting when resources are limited for long-term storage. PMID- 15350459 TI - The endothelium and early immune activation: new perspective and interactions. AB - Far from being just a membrane for the passive transport of cells, molecules, and water between blood vessels and their interstitium or a passive target for humoral immunological reactions, the endothelium is now being viewed as an active modulator of both normal physiological homeostasis and the early inflammatory response. The repertoire of receptors and mediators produced by endothelial cells overlap those traditionally assigned to the immune system. This new paradigm together with the changes caused by brain death in the cadaver donor have far reaching repercussions on how rejection is conceived, while opening new venues for its prevention. The respective theoretical roles of the neuro-endocrine immunological axis and of syndecans are highlighted. PMID- 15350460 TI - Xenotransplantation of human decay accelerating factor transgenic porcine kidney to non-human primates: 4 years experience at a Canadian center. AB - From July 1999 to December 2002, a total of 27 baboons underwent transplantation with human decay accelerating factor (hDAF) transgenic porcine kidneys. The immunosuppressive protocol included combinations of 1 or more of the following immunosuppressive agents: cyclosporine, TPC, cyclosphophamide, methylprednisolone, GAS914 (a polymeric alpha-Gal), rabbit antithymocyte serum (RATS), and rapamycin derivative (RAD). The animals were followed up clinically with monitoring of renal function based on daily creatinine and urinary output. Histopathological examination of the tissue samples was performed after paraffin embedding. Electron microscopy was performed on occasion. All pathology was read blindly with a semiquantitative scoring method to assess the presence or absence of histological features of xenotransplant rejection. Mean survival time was 20.7 days (range, 4-75 days) with 15 animals dying of classic humoral rejection and 12 dying of complications unrelated to rejection. The most frequent complications were as follows: chronic diarrhea with malabsorption, bleeding diathesis, infections, and drug toxicity. Irrespective of the immunosuppressive regimen, a significant number of baboons (10) developed untreatable diarrhea and gastrointestinal bleeding. Samples of their gastrointestinal tracts showed evidence of varying degrees of necrotizing vasculitis with immunoglobulin (Ig)G,IgM, and fibrin deposition. The survival in pig-to-baboon kidney xenotransplantation continues to be limited by acute xenograft rejection and complications of immunosuppression. Although significantly increased survivals have been obtained, an ideal protocol has not yet been achieved. Efforts will continue in an attempt to develop a protocol with minimal complications and long term survival beyond 90 days. PMID- 15350462 TI - Synthetic peptides mapping to epitopes of the extracellular domain of the interleukin-2 receptor beta-chain to inhibit T-cell activation. AB - Expression of the high-affinity IL-2 receptor (IL-2R) on the surface of activated T cells makes it an attractive target for selective inhibition of alloreactive T cells in organ transplantation. IL-2 binds to its receptor via the extracellular domain of the beta-chain. In this study we synthesized synthetic peptides that map to epitopes of this domain and tested their ability to inhibit the activation and proliferation of mitogen-stimulated peripheral-blood T cells. Solid-phase synthesis was applied to create three oligopeptides of primary structures corresponding to the epitopes M(107)-E(118), Y(178)-Q(199), and E(190)-Q(199) of the extracellular domain of the IL-2Rbeta-chain. A nonhomologous peptide served as control. Peripheral-blood mononuclear cells isolated from 16 healthy volunteers (median age 41 years, range 26 to 56 years) were cultured with various concentrations of peptides and the mitogen phytohemagglutinin (PHA). The inhibitory effect of the peptides on cell proliferation was evaluated by automated cell counting and colorimetric proliferation assays. Cell activation was assessed by immunophenotyping using antibodies directed toward CD4, CD25, or CD69. The amount of IL-2 in culture supernates was measured by enzyme-linked immunoassay. Cultures in the presence of the peptide M(107)-E(118) (500 nmol/L) inhibited PHA-induced T-cell proliferation by 38%, and IL-2 secretion by 57%. Immunophenotyping confirmed suppression of activated T cells. Peptides Y(178) Q(199) and E(190)-Q(199) inhibited proliferation, but failed to significantly affect IL-2 secretion. The control peptide showed no effect on the activation parameters. Our data indicate that the M(107)-E(118) peptide has promise for organ transplantation therapy. PMID- 15350463 TI - Humoral sensitization against rejected grafts: specific antibodies to graft immunogenic amino acid triplets. AB - Humoral sensitization against immunogenic amino acid (aa) triplets expressed on a rejected graft was analyzed in 83 retransplant candidates. All patients had lost a graft with HLA-A,-B mismatches. The alloantibodies were detected by a complement-dependent cytotoxicity (CDC) technique and an ELISA method in parallel; they were classified as HLA graft-specific (GS) and non-GS antibodies. The aa triplet specificity of the antibodies was assessed using the HLAMatchmaker algorithm. HLA class I antibodies were detected in 74 of 78 (94%) cases, including GS reactivity in 55 (74.3%) and non-GS in 72 (97.2%), either alone (n = 19) or in parallel with GS antibodies (n = 53). For all HLA-GS-antibody-reactive patients, we defined the specificity against immunogenic aa triplets on the previous graft. Moreover, antibodies specific to graft aa triplets were observed within the non-GS antibodies among 19 of 19 and 28 of 53 cases, respectively. Therefore, aa triplet-specific antibodies against the rejected graft were present in all 74 cases with HLA class I antibodies. Antibodies against aa triplets expressed on all HLA class I-mismatched graft antigens were present in 73% of cases. The high extent of humoral alloreactivity against a rejected graft supports the decision to avoid repeated exposure to immunogenic aa triplet mismatches on a second graft. An accurate analysis for performed antibodies in these cases may be beneficial to select the most suitable second donor. PMID- 15350464 TI - HLA class I donor-specific triplet antibodies detected after renal transplantation. AB - The purpose of this study was to investigate whether IgG, non-donor-specific anti HLA class I antibodies (HLAabI) detected after renal transplantation recognize immunogenic amino acid triplets expressed on the foreign graft. In addition, we sought to evaluate the effect of these antibodies as well as other posttransplant HLAabI on graft outcome. Posttransplant sera from 264 renal recipients were tested for the presence of IgG HLAabI and HLA class II-specific alloantibodies (HLAabII) by ELISA. The HLAMatchmaker computer algorithm was used to define the HLA class I non-donor-specific antibodies, which seem to recognize immunogenic amino acid triplets. Donor-specific triplet antibodies (DSTRab) were detected in 16 of 22 (72.7%) recipients based on at least one HLA-A or -B mismatched antigen with the donor. DSTRab were found either without (n = 7) or with (n = 9) HLA donor-specific antibodies (HLA-DSA). The presence of DSTRab alone in the periphery was associated with acute rejection, whereas the presence of both DSTRab and HLA-DSA was associated with chronic rejection and graft failure. PMID- 15350465 TI - Minor histocompatibility antigen HA-1 and HPA-5 polymorphisms in HLA-identical related bone marrow transplantation. AB - The minor histocompatibility antigens (mHags), HA-1 and HPA-5, are immunogenic alloantigens shown to be responsible for graft-versus-host disease (GVHD) in HLA identical bone marrow transplantation. Both antigens have two known alleles each, resulting in a single amino acid polymorphism. The HA-1H allele encodes histidine, whereas the HA-1R allele encodes arginine. The HPA-5b (Br(a)) allele encodes lysine, whereas the HPA-5a (Br(b)) encodes glutamic acid. In this study, 49 bone marrow transplant recipients and their genetically related HLA-identical donors were evaluated for the presence of HA-1, whereas 39 recipients, different from the abovementioned ones, and their HLA-identical siblings were analyzed for the presence of HPA-5. The frequencies of the two alleles of HA-1 in the recipient population were HA-1R = 0.663 and HA-1H = 0.336. In the donor population, the respective frequencies were 0.704 and 0.296. Seven donors (14.5%) were mismatched with the recipients for HA-1H. In contrast, the frequencies of the two alleles of HPA-5 in the recipient population were HPA-5a = 0.859 and HPA 5b = 0.141; whereas, among donors, they were 0.820 and 0.180, respectively. Five donors (12.8%) were found to be mismatched with their recipients for HPA-5. These results provide insight into the polymorphism of mH antigens based on the study of their frequencies in bone marrow transplant recipients and their genetically HLA-identical siblings, an endeavor that is essential to investigate the presence of HA-1 and HPA-5 mHags. PMID- 15350466 TI - Use of mycophenolate mofetil in the experimental allo-transplantation of hepatocytes in rats with fulminant hepatic failure. AB - AIM: The aim of the study was to evaluate the beneficial effect of mycophenolate mofetil (MMF) as an immunosuppressive agent for experimental transplantation of hepatocytes in rats with fulminant hepatic failure (FHF). MATERIALS AND METHODS: Six Wistar rats were used as donors and 40 Lewis rats at recipients, including four groups of 10 animals each. Group A received no treatment; Group B, cyclosporine (20 mg/kg days 0-5 and 10 mg/kg days 6-15); Group C, MMF (12 mg/kg per os every day); and Group D, MMF (23 mg/kg per os every day). Hepatocytes were transplanted intrasplenically. Animals were followed for 15 days. RESULTS: The survival rates for Group A were maximum 72 h, whereas Groups B, C, and D showed 50%, 70%, and 80%, respectively. Biochemical evaluation and histology showed satisfactory function and engraftment, respectively. CONCLUSION: The use of MMF in this experimental model yielded safe, satisfactory immunosuppression especially at the dose of 23 mg/kg. PMID- 15350467 TI - Hepatocyte function during experimental use of a bioartificial liver. AB - The aim of the present study was to compare the function of fresh versus cryopreserved hepatocytes in an experimental bioartificial liver system (BAL), especially designed to reproduce clinical parameters. Our BAL consists of a pump, a plasma reservoir, a membrane oxygenator, and a hollow fiber module loaded with 5 x 10(9) isolated porcine hepatocytes, either fresh (n = 5) or cryopreserved (n = 5). In the present setting, the system was isolated and perfused for 6 hours with recirculating plasma obtained from pigs with ischemic liver failure (toxic plasma). The following parameters were studied at 0 and 6 hours: oxygen consumption by the hepatocytes in the bioreactor, hepatocyte viability, as well as plasma concentrations of AST, LDH, ammonia, urea, and total bilirubin. MEGX concentrations were measured following injection of lidocaine into the system 30 minutes after initiation of plasma recirculation. Compared to cryopreserved cells, fresh hepatocytes showed higher viability at both time points studied (P <.05). Furthermore, during BAL sessions, ammonia levels were reduced while urea, AST, and LDH levels were increased with both preservation types (P <.05). Total bilirubin levels increased only during sessions with cryopreserved hepatocytes. After lidocaine administration, both fresh and cryopreserved hepatocytes were capable of producing MEGX; however, fresh-cell bioreactors produced significantly more MEGX at both 30 and 60 minutes after lidocaine administration. Oxygen consumption was significantly higher by fresh-cell bioreactors both before and after BAL use. In conclusion, hepatocytes in the BAL bioreactor showed preservation of important metabolic functions, when perfused with homologous toxic plasma. Fresh cells appeared to respond better than did cryopreserved ones. PMID- 15350468 TI - The impact of thymoglobulin on renal function and calcineurin inhibitor initiation in recipients of orthotopic liver transplant: a retrospective analysis of 298 consecutive patients. AB - BACKGROUND: Renal dysfunction remains the Achilles' heel of calcineurin inhibitor (CI)use. The purpose of this study was to assess our institutional, renal-sparing strategy using thymoglobulin (TMG) in recipients of orthotopic liver transplants. METHODS: We performed a retrospective analysis of data from 298 adult recipients who were transplanted between 1991 and 2002. The patients were divided into two groups: those induced with TMG (group 1) and those that were not treated with this agent (group 2). A subgroup analysis was performed of patients with baseline serum creatinine values above 1.5 mg/dL (group 1A received TMG; group 2A did not). All patients received tacrolimus or cyclosporine (CyA) maintenance immunosuppression. RESULTS: Indications and demographics were similar between the two groups. Although there was no difference in patient and graft survivals, there was a statistically significant benefit in the rejection-free graft survival at 1 year for group 1 (51% vs 39%; P =.02). Furthermore, serum creatinine at 6 months was lower for group 1, despite a similar baseline creatinine. Subgroup analysis for patients with baseline abnormal serum creatinines showed that group 1A displayed an improved rejection-free graft survival at 1 month but not at 1 year. CONCLUSIONS: Thymoglobulin induction therapy may allow a delay in the initiation of CI therapy without compromising patient and graft survival, while preventing early rejection, even among patients with baseline renal dysfunction. PMID- 15350469 TI - Cyclosporine levels at 2 hours after dose and body mass index in relation to graft function in renal transplant patients treated with azathioprine or mycophenolate mofetil. AB - The aim of this study was to evaluate the effect of C(2) levels on renal graft function in relation to body mass index (BMI). This retrospective study of 95 renal transplant patients included 53 on AZA and 42 on MMF at 3.1 years after transplantation. The cohort was divided into groups according to their C(2) levels, namely <600 ng/mL, 600 to 900 ng/mL, or >900 ng/mL, and according to BMI (>26 kg/m(2)). In every group, we evaluated the percentage of patients with an increase in creatinine by 1 mg/dL or >/=50% from the first year posttransplant. There was no difference in age, gender, graft source, and dose of corticosteroids or CsA between the groups. Patients on AZA with C(2) 600 to 900 ng/mL showed a lower prevalence of renal dysfunction (3.4%) than those with C(2) levels <600 ng/mL (14.3%) or >900 ng/mL (20%). Seventeen percent of the patients on AZA and 11.9% on MMF had BMI >26 kg/m(2) (P = NS). An increased serum creatinine was present in 22.2% of patients with BMI >26 kg/m(2) in the AZA group vs 20% in the cohort MMF (P = NS). These findings suggest that long-standing renal recipients on AZA with C(2) levels of between 600 and 900 ng/mL show better preservation of renal function. We did not identify differences on the basis of C(2) levels in MMF-treated recipients. The influence of BMI on long-term graft function seemed to be independent of AZA or MMF therapy. PMID- 15350470 TI - Glomerular filtration rate estimation in renal transplant patients based on serum cystatin-C levels: comparison with other markers of glomerular filtration rate. AB - PURPOSE: The assessment of glomerular filtration rate (GFR) is the most commonly used test of renal function. Cystatin-C, a cysteine protease inhibitor, which can be measured by light-scattering immunoassay, possesses many of the attributes required of the ideal GFR marker. Conversely, many endogenous markers that are widely used for the estimation of GFR such as serum creatinine (SCr) are not ideal. The present study was undertaken to evaluate the clinical application of serum cystatin-C (CysC) as a new marker of GFR in renal transplant patients. METHODS: Eighteen patients (9 men) were enrolled in the study (mean age: 46.35, range: 31-67 years) to measure serum CysC levels and compare them, with SCr, creatinine clearance (CCr), as well as the Cockcroft-Gault equation (CG) or the MDRD as indicator of GFR. Spearman's correlation coefficient was used to determine the relationship between CysC and other markers. RESULTS: There was a significant negative correlation between serum CysC and CCr (r = -0.768). Moreover, the CysC level was negatively correlated with CG (r = -0.854), positively correlated with SCr (r = 0.629), and negatively correlated with MDRD (r = -0.604). CONCLUSIONS: These results indicate that measurement of serum cystatin-C was useful and accurate to estimate GFR in renal transplant patients. The recent literature confirms our data although there are concerns about nonrenal influence on this test. Although serum CysC can generally be recommended as a marker for GFR, our study is still in progress seeking to validate the conclusions in a larger number of patients. PMID- 15350471 TI - Leukoreduction and acute rejection in liver transplantation: an interim analysis. AB - BACKGROUND: Little is known about the effect of blood transfusions and leukoreduction on acute rejection in liver transplantation. The purpose of this study was to assess the impact of leukoreduction on the occurrence of early rejection episodes in liver transplantation. METHODS: In 1999, mandatory leukoreduction was implemented in our program. Data from 339 consecutive liver transplant recipients were analyzed with attention to the time period as a proxy for leukoreduction, the number of transfusions, the wait list status, the hepatitis B or C status, the recipient age, and the type of immunosuppression. RESULTS: Using an early (6-month) rejection-free graft survival model, we observed that introduction of leukoreduction was independently associated with fewer rejection episodes (P =.001). Despite the lower rejection rate, due to a regimen of tacrolimus and antithymocyte globulin, the effect of implementation of leukoreduction remained significant (P =.021). CONCLUSION: The use of leukoreduction is associated with fewer early rejections, irrespective of the type of immunosuppression. These data support an exploration of the immunomodulatory effect of leukoreduction. PMID- 15350472 TI - Extracorporeal membrane oxygenation circulatory support after cardiac surgery. AB - OBJECTIVES: Postcardiotomy acute severe heart failure cannot be managed by medical treatment alone and most often requires some form of mechanical support. In this study we evaluate the efficacy of postoperative extracorporeal membrane oxygenation (ECMO) support following surgery for congenital heart disease (CHD) in infants and children. METHODS: Over a 6-year period from October 1997 to October 2003, 10 patients aged 5 days to 28.5 months (median 3 months) who underwent surgical procedures for CHD received postoperative mechanical support for failing cardiac function despite optimal medical therapy. In 3 patients ECMO was instituted in the operating room (OR) and in 7 patients this was introduced in the intensive care unit (ICU) 2 to 48 (median 20) hours postoperatively. RESULTS: Four patients (40%) were successfully weaned, while support was withdrawn in the remaining 6 due to irreversible vital organ damage. Following successful weaning, one of the survivors died 8 hours later from barotrauma and intrapulmonary hemorrhage, and another died 4 months later from persistent heart failure. The other two patients remain well in NYHA class II. CONCLUSIONS: Despite the adverse effects of ECMO, the methodology provided the necessary support and allowed the failing heart to recover in a number of patients where inotropic support alone proved inadequate. PMID- 15350473 TI - Correlation of cell necrosis and tissue calcification with ischemia/reperfusion injury after liver transplantation. AB - BACKGROUND: The cellular events following liver ischemia/reperfusion (I/R) during transplantation are largely unknown. The spectrum of I/R damage to the liver can be clinically revealed by the development of primary graft dysfunction or nonfunction. Because viral-induced liver necrosis has been associated with the development of calcifications in an animal model, we investigated the spectrum of I/R changes identified at an ultrastructural level among livers after liver transplant (LT). MATERIALS AND METHODS: Random liver biopsies from five recipients with different degrees of liver dysfunction (LD) were processed for light (LM) and electron (EM) microscopic examination. The degree of calcification was estimated as mild-moderate or severe. The degree of cell vacuolization, used as a surrogate marker of cell necrosis, was reported as mild-moderate or severe. RESULTS: Two patients with severe LD had obvious calcifications by LM and EM examinations. Both showed significant vacuole formation, suggesting a severe degree of cell necrosis, and both succumbed to the sequelae of their LD. One patient showed evidence of mild calcifications at EM (but not LM) examination, with mild vacuole formation. The remaining two patients displayed no microcalcifications. Both presented only mild vacuole formation. Both patients recovered from LD and are currently alive. CONCLUSION: In this preliminary report, we conclude that the clinically observed degree of LD after orthotopic liver transplant (OLT)correlates well with ultrastructural modifications. These include calcification and vacuole formation. We believe that both findings can be used as surrogate markers of a clinically significant hepatic I/R injury. PMID- 15350475 TI - Renal failure in Yemen. AB - Renal failure remains a serious cause of mortality in Yemen. Our region has 1.25 million population and our hospital is the central hospital, which has a nephrology department and performs dialysis for the region. Between January 1998 and December 2002, we admitted 547 patients; including children, with acute renal failure (ARF) and chronic renal failure (CRF). CRF was observed in 400 patients, an incidence of 64 per million per year and a prevalence of 320 per million. ARF occurred in 147 persons with an incidence of 23.5 per million per year and a prevalence of 117.5 patients per million. Of all patients, 72% were adults (age range, 20-60 years) with a male preponderance. As a tropical country, malaria (27.9%), diarrhea (13.6%), and other infectious diseases were the main causes. Next most common were obstructive diseases causing CRF and ARF (26.8% and 12.9%, respectively), mainly urolithiasis, Schistosomiasis, and prostatic enlargement. However the cause of CRF in 57.5% of patients was unknown as most persons presented late with end-stage disease (64.7%), requiring immediate intervention. Other causes, such as hepatorenal syndrome, snake bite, diabetes mellitus, and hypertension, showed low occurrence rates. Patients presented to the hospital mostly in severe uremia and without a clear history of prior medications. The major findings were vomiting, acidosis, and hypertension with serum creatinine values ranging between 2.8-45 mg/dL (mean value, 13.4 mg/dL). Anemia was observed in 80.4% of CRF versus 62.6% of ARF patients. Hypertension prevalence was 65.5% among CRF patients, of whom 25% were in hypertensive crisis, whereas among ARF the prevalence was only 26.5%. PMID- 15350476 TI - Pattern of acute renal failure in a tertiary hospital in the United Arab Emirates. AB - Acute renal failure (ARF) is a challenging problem in nephrology. To evaluate the pattern, management and outcome of ARF in our tertiary hospital, we analyzed the data of all 81 patients admitted with or developing ARF in hospital between January 2002 and June 2003. The 45 men and 36 women of mean age 56.2 +/- 21 (range 13 to 91) years were managed either on the ward (n = 48; 59%) and or in the ICU (n = 33; 41%) 10% were direct admissions to the nephrology service with ARF, and 90% developed ARF in hospital. Thirty percent were referred by oncology services and 15% by general medicine. Sepsis was the cause of ARF in 36 (44%) patients, followed by drug nephrotoxicity in 11 (14%), and obstructive uropathy in 9 (11%). Comorbid conditions were hypertension in 28 (35%); diabetes in 27 (33%); chronic renal failure, 19 (23%); ischemic heart disease 19 (23%); and liver disease 12 (15%). The most common predisposing factor was hypotension in 42 (52%), dehydration in 32 (40%), and drug nephrotoxicity in 20 (25%). Sixty patients (74%) were managed conservatively, and 21 (26%) required renal replacement therapy. The length of hospital stay was 29.5 +/- 38.4 (range 2 to 279) days. Patient survival for those managed on the ward was 71% compared to 33% for ICU patients (P <.00001). Renal survival was 83% for ward patients, compared to 48% for those in the ICU (P <.001). This study showed that majority of ARF developed in-hospital with oncology patients constituting the greatest proportion. Sepsis was the leading cause of ARF and hypotension, the main predisposing factor. Patients treated in the ICU showed a worse prognosis for both patient and renal survival. PMID- 15350477 TI - Outcome and predictive factors of acute renal failure in the intensive care unit. AB - The development of acute renal failure (ARF) in the ICU setting carries a high morbidity and mortality. To assess the outcomes and its predictive factors in our ICU, we analyzed the data of patients with ARF treated during 18 months. The 33 patients included 21 men and 12 women of mean age 51 +/- 21.7 years (13 to 87). Sepsis with multi-organ dysfunction (MOD) was the leading cause of ARF (58%). Comorbid conditions were malignancy in 30% of patients, diabetes mellitus in 24%, hypertension in 21%, ischemic heart disease in 21%, liver disease in 15%, and chronic renal failure in 15%. Predisposing factors were hypotension in 67% of cases, dehydration in 36%, drug related in 33%, congestive heart failure in 24%, and liver cirrhosis in 6%. Twenty-five (76%) patients needed mechanical ventilation, 22 (67%) were anuric, 18 (55%) had MODS, and 15 (45%) needed inotropic support. Length of stay in hospital was 27.2 +/- 28.0 days (2 to 94). Nineteen patients (58%) were managed conservatively and 14 (42%) by renal replacement therapy. Patient mortality was 67% and renal mortality 52%. The impact of the following factor: was assessed on patient and renal outcome was assessed ventilation support, presence of oliguria, need for inotropes, and presence of MOD. Patient mortality was significantly influenced by an elevated odds ratios (OR) (95% CI): mechanical ventilation [OR = 34 (95% CI 1.95 to 538)], and presence of MODS [OR = 12.3 (95% CI 2 to 75)]. Renal mortality was influenced by mechanical ventilation [OR = 12.3 (95% CI 1.6 to 119)], oliguria [OR = 12 (95% CI 2 to 72)], inotrope support [OR = 10 (95% CI 2 to 52), and MOD [OR = 35 (95% CI 3.5 to 35.0)]. This study confirms the high patient and renal mortality of ARF among patients to ICU. The four parameters were excellent predictors of renal outcome, while only the need for mechanical ventilation and the presence of MOD were predictors for patient survival. PMID- 15350478 TI - End-stage renal disease in Kuwaiti children: an 8-year experience. AB - BACKGROUND: Prior to the establishment of the pediatric nephrology service in Kuwait in 1995, no accurate registry of end-stage renal disease in children was available due to management by various adult nephrologists. In this study we analyzed our experience with renal replacement therapy in children, as the only center in the country offering this service for the past 8 years. SUBJECTS AND METHODS: The records included all children less than 16 years of age with end stage renal disease treated in the pediatric nephrology unit over a period of 8 years (January 1995 to December 2002). RESULTS: Of the 48 children boys comprises 52% and the overall mean age at institution of dialysis was 94.4 months. Causes of renal disease included congenital structural anomalies in 52%, including obstructive uropathy in 16.6%, vesicoureteric reflux in 16.6%, and renal dysplasia/hypoplasia in 18.7%. Hereditary nephropathy was diagnosed in 35.4%, including primary hyperoxaluria in 10.4%, nephronophthisis in 2%, autosomal recessive polycystic renal disease in 8%, and glomerulopathies in 14.5%. Other etiologies constituted 14%. Renal replacement therapy was necessary in 43 patients: 46% by peritoneal dialysis and 43% by hemodialysis. The mortality rate in the dialyzed group was 16%. Twenty-four patients received kidney transplants from, cadaveric donors in 19 cases. CONCLUSION: Genetic factors contributed to the high incidence of end-stage renal disease, which is most likely due to the common practice of consanguineous marriages in our country. PMID- 15350479 TI - The spectrum of glomerular diseases in the kingdom of bahrain: an epidemiological study based on renal biopsy interpretation. AB - Glomerular diseases continue to be the leading cause of end-stage renal disease globally. Hence it is important to recognize the glomerular disease pattern in any given geographical area to understand the pathobiology in the region as well as the incidence and progression of the disorder. A total of 498 renal biopsies were performed on patients with proteinuria, hematuria, and mild to moderate renal impairment during a period of 13 years (between January 1990 and December 2002) at a tertiary care hospital. Primary glomerular disease accounted for two thirds of the glomerular diseases, which was 44.8% of all renal biopsies. The most common histological lesion was minimal change disease (30%). Focal segmental glomerulosclerosis was the second most common lesion (23.8%) followed by membranoproliferative glomerulonephritis (14.3%). Secondary glomerular disease included 33.6% of glomerular diseases with 22.7% with lupus nephritis as the commonest lesion (38.9%) followed by diabetic nephropathy (31.9%) and hypertension (20.4%). Tubulointerstitial diseases accounted for 13.1% of all renal biopsies, whereas transplant diseases were noted in 12.2%. The miscellaneous group including inadequate biopsies, which constituted 7.2% of all the tissues. The results of this analysis were compared with surveys from other parts of the world. PMID- 15350480 TI - Renal amyloidosis followed more than 5 years: report of 12 cases. AB - Renal involvement with amyloidosis is common but causes patient survival to be poor, rarely reaching 5 years. In this study, we retrospectively reviewed clinical and biological characteristics as well as treatments and outcomes of patients with renal amyloidosis followed for more than 5 years. Between 1975 and 2003, 485 patients were diagnosed with renal amyloidosis including only 12 patients who were followed more than 5 years. The six men and six women of mean age 42.4 years (range 18 to 66 years) displayed renal signs of lower limb edema in all cases; hypertension in four cases, proteinuria on urinalysis in all cases with microscopic hematuria in five cases. Biological tests showed nephrotic syndrome in 11 patients, normal renal function in nine patients, and renal failure in three patients whose mean creatinine was 481.6 micromol/L (range 294 to 726). The amyloidosis was AA type in 11 cases and non-AA in one case. An etiologic survey revealed spondylarthropathy in one patient, pulmonary tuberculosis in two patients, chronic bronchitis in three patients, hepatic hydatic cyst in one patient, Mediterranean familial fever in two patients, Crohn's disease in one patient, Hodgkin's lymphoma in one patient, and multiple myeloma in one patient. Specific treatment was initiated with colchicine in seven patients. At a 110-month mean follow-up (range 53 to 153 months), remission of nephrotic syndrome was observed in four cases, progression to chronic renal failure in two patients, and to end-stage renal failure in five cases (range 53 to 196 months), with stabilization of renal function in seven patients. In conclusion, primary amyloid disease should be optimally suppressed in patients with renal involvement. The role of this treatment in remission of renal amyloidosis is not well established. This efficacy of the treatment has been demonstrated in some patients with improved survival. PMID- 15350481 TI - A multicenter clinical trial in the Arab world. AB - Financial, social, and perhaps religious, ethical, and other factors hinder investment and performance of clinical research in Arab countries. We report in that conducting multicenter clinical trials is feasible in the Arab world, describing our experience in planning and conducting one of the first multicenter, multinational, clinical trials in the region. A multicenter clinical trial sought to document the efficacy and safety of Epotin for the treatment of anemia in patients on maintenance hemodialysis. Among 29 centers contacted for participation, a positive response was obtained from 17 (62%) located in eight countries. The initial recruitment period of 3 months was extended to 1 year. Among the participating centers, 16 forwarded their results in time, with one being late. There were minor and a few major protocol violations, the latter requiring exclusion of data from the final analysis. Sponsorship was mainly by a local pharmaceutical company (Julphar). A co-coordinating body was crucial to trace and gather the data. Since conduct of the trial required considerable time and effort from investigators, the use of modern information technology could have reduced the effort and improved the outcomes. We conclude that multicenter clinical trials, which are essential can be conducted in this region. This experience needs to be repeated and refined. PMID- 15350482 TI - The new rHuEPO alpha (epotin) in the management of anemia of end-stage renal disease in patients on maintenance hemodialysis. AB - Recombinant human erythropoietin has proved to be effective to treat anemia of end-stage renal disease (ESRD). The aim of this study was to assess the efficacy and safety profile of Epotin, a rHuEPO produced in the Middle East. One hundred thirty patients with Hct 1000 pg/mL associated with severe osteitis fibrosa, debilitating pruritus, marked soft tissue calcification, or hypercalcemia with hyperphosphatemia, which sometimes complicated vitamin D therapy. All patients were resistant to more conservative measures, including control of serum phosphate, attention to oral intake and dialysate calcium levels, and oral/intravenous administration of active vitamin-D pulse therapy. Ultrasound and technetium 99-sestamibi scan were used to image the thyroid and the parathyroid glands. Total PTX with autotransplantation was performed in 23 patients; subtotal PTX was performed in 3 patients. Histology of frozen sections taken intraoperatively showed nodular changes in 14 and diffuse hyperplasia in 12 cases. During the 2-year follow-up period significant reductions in parathyroid hormone, alkaline phospatase blood levels, skeletal changes, and soft tissue calcifications were observed. Pruritus improved in half the cases. Some improvement in hemoglobin and hematocrit was also noticed. The complication rate after PTX was low. Transient postoperative hypocalcemia requiring intensification of calcium and vitamin D therapy was seen in cases with high preoperative alkaline phosphatase levels. Recurrence was observed in two cases. Hypoparathyroidism was not recorded. We conclude that surgical reduction of parathyroid mass is a safe and effective treatment for symptomatic disease not suppressible by pharmacologic means. PMID- 15350485 TI - The role of ambulatory blood pressure measurements in adolescence and young adults. AB - INTRODUCTION: Ambulatory blood pressure measurements (ABPM) are currently used for the diagnosis and treatment of hypertension. It is widely recognized that the casual/clinic blood pressure is less representative of the true blood pressure than the average ABP. METHODS: Among the 15 records for 14 patients analyzed, 80% were obtained from type I diabetics. The overall age, ranged between 11 and 23 years (mean = 17.2 years). ABPM was performed using the Quiet Trak 98 model by Welch Allyn automatic auscultatory sphygmomanometer for a 24-hour period, for those with clinic documented hypertension. The BP was considered high if recorded as systolic > 140 and diastolic > 90 for the whole period or for period I (7 am to 11 pm), and >120 systolic and >85 diastolic for period II (11 pm to 7 am), or >130 systolic and >85 diastolic for diabetic patients. RESULTS: The BP was recorded as normal in 20% of the patients; in 33% it was normal during the daytime but high during the nighttime, and 80% of the diabetic patient were found to be nondippers during night time. CONCLUSION: ABPM is helpful to exclude White coat hypertension and nondippers especially among diabetic patients. PMID- 15350486 TI - Impact of initial blood pressure on the mortality of diabetics undergoing renal replacement therapy. AB - The aim of this study was to determine the blood pressure (BP) profiles and their impact on mortality among a cohort of uremic diabetics treated by hemodialysis. The studied population includes all type II diabetics starting hemodialysis for end-stage renal disease between 1990 and 1996. There were 221 patients (144 men, 77 women) aged from 37 to 78 years, were all followed until death or December 2003 without any censored data. Survival analysis to identify predictors of death was performed using the actuarial method, Cox proportional model, including systolic, diastolic, mean, and pulse blood pressures (SBP, DBP, MBP, PP). One hundred seventy-eight patients (80.5%) were hypertensive at the start of dialysis. Hypertension preceded the diagnosis of diabetes in eight cases (4.5%); 154 patients (86.5%) received antihypertensive drugs and only 23 (14.9%) had well controlled hypertension. Our population was subdivided into four groups according to their BP levels at the time of beginning of dialysis; G1 (19.5%): normal BP (SBP [90 to 140] and DBP [60 to 90]); G2 (30.3%): Hypertension stage 1 (SBP [140 to 160] and/or DBP [90 to 100]); G3 (32.1%): hypertension stage 2 (SBP [160 to 180] and/or DBP [100 to 110]); G4 (18.1%) hypertension stage 3 (SBP [180 to 220] and/or DBP [110 to 120]). Mean age and comorbidities were similar among the four groups. During a cumulative follow-up period of 872 patient-years, 191 patients died, representing a rate of 21.9 per 100 patient-years; 20.42% of these deaths occurred during the first 3 months of dialysis. Normotensive patients showed lower survival rates without any significant difference in comparison with those of other hypertensive groups. None of the initial BP parameters (SBP, DBP, PP, MBP, hypertension stages) seemed to influence early or global mortalities, which were rather related to the urgent onset of renal replacement therapy, to age, to serum albumin, and to the score of associated morbidities. We conclude that mortality of our hemodialyzed diabetics was not influenced by the blood pressure parameters recorded at the onset of dialysis. PMID- 15350487 TI - Seroprevalence of hepatitis C virus and hepatitis B virus among dialysis patients in Bahrain and Saudi Arabia. AB - Dialysis patients are at risk for contracting blood-borne infections, including hepatitis viruses (HBV and HCV). The aim of this study was to assess the prevalence of HBV and HCV infection among hamodialysis patients in Bahrain and Saudi Arabia. Study subjects comprised 81 Bahraini and 34 Saudi dialysis patients, and as control 7714 Bahraini and 2330 Saudi blood donors. Serologic markers of HBV (HBsAg, anti-HBc) and HCV (anti-HCV) were determined by EIA and confirmed by PCR (HBV) and RT-PCR (HCV). Higher prevalence of HCV (9.240% vs 0.300%, P <.001), HBsAg (5.88% vs 0.620%; P <.001), but not anti-HBc (1.7% vs 4.6%; P =.01) were seen in patients compared to controls, respectively. When compared to Bahrainis, higher prevalence of HBsAg (11.8% vs 3.7%) and anti-HCV (14.7% vs 7.4%) were seen among Saudi patients, respectively. Double HCV infection was frequent, and the most prevalent types were HCV1a/1b plus HCV4 in Bahraini, and HCV 2/2a plus HCV 4 among Saudi dialysis patients. Our results are the first report on viral hepatitis among dialysis patients in Bahrain, and the first to compare HBV/HCV rates among dialysis patients in the Eastern Arabian peninsula, and confirms other results that documented increased HBV and HCV infection among dialysis patients. Future studies aimed at assessing the status and to monitor the progress of viral hepatitis infection among dialyzed and transfused patients will have a strong impact on patient diagnosis, follow-up, and treatment. PMID- 15350488 TI - Profiled hemodialysis reduces intradialytic symptoms. AB - A prospective study on the effect of profiled hemodialysis on intradialytic symptoms was undertaken among patients recruited between September 2002 and December 2002. Evaluated intradialytic symptoms included hypotension, muscle cramps, dizziness, headache, nausea, discomfort, thirst, and shortness of breath. Symptomatic patients were allocated to one mode of combined sodium and ultrafiltration profile during hemodialysis. The programs were readily available on Fresenius MC 4008 H&E hemodialysis machines. Evaluation was performed before profiling and at 2, 4, and 6 weeks into the profiled hemodialysis. On standard hemodialysis 40 (36.4%) patients were symptomatic. Hypotension was reported in 29 (72.5%) of patients receiving standard treatment. Dizziness, headache, and muscle cramps were reported in 22 (55%), 15 (37.5%), and 9 (22.5%) patients, respectively. These symptoms were significantly (P <.05) improved at 2, 4, and 6 weeks of profiling. Other symptoms, such as discomfort, nausea, vomiting, and thirst, were infrequently reported among patients without or with profiling. There was no significant difference between the applied profiles when compared with each other. In conclusion, sodium and ultrafiltration profiling are effective techniques to reduce acute adverse side effects of hemodialysis. They improve patient well-being and dialysis tolerance, minimize interventions during dialysis. PMID- 15350489 TI - Baseline cardiac troponin I in patients on maintenance dialysis. AB - INTRODUCTION AND OBJECTIVES: There have been conflicting reports in the medical literature regarding baseline levels of cardiac troponins and their usefulness as specific markers of acute myocardial injury in patients on maintenance dialysis. Hence we undertook a study of the normal range of baseline cardiac troponin I (cTnI) in patients on maintenance dialysis in our center. METHODOLOGY: Seventy five patients aged 45 years or more on maintenance dialysis for at least 1 month were included in the study. None had had an acute cardiac event requiring admission to the hospital during the preceding month. Samples of blood were collected and cTnI levels were estimated using an immunoenzymatic assay (Access Immunoassay system, Beckmann). Correlation of cTnI with gender, age, body mass index (BMI), and adequacy of dialysis (Kt/v) were also done. RESULTS: Forty-six (61.3%) patients were on maintenance hemodialysis and 29 (38.6%) were on peritoneal dialysis. Twenty-four (32%) were over the age of 65 years and 37 (49.3%) were men. Levels of cTnI ranged from 0.000 to 0.568 ng/mL, with mean of 0.040 +/- 0.080. Only six (8%) had levels above 0.1 ng/mL, which was the cut off level for significance in our laboratory. There was no statistically significant correlation of cTnI levels with gender, age, BMI, or Kt/v. CONCLUSIONS: Baseline cTnI levels were below significant levels in the majority of maintenance dialysis patients aged at least 45 years or more. Hence cTnI can be used as a valid marker of acute myocardial injury in this group of patients. PMID- 15350490 TI - Alpha-interferon with ribavirin in the treatment of hemodialysis patients with hepatitis C. AB - INTRODUCTION: Hepatitis C Viral (HCV) infection is the leading cause of chronic liver disease in end-stage renal disease patients (ESRD). The impact of HCV on patient and graft survival posttransplantation is controversial. The most successful approach is to eliminate the virus while the patient is on dialysis prior to transplantation. The main aim of this pilot study was to assess the efficacy of combined alpha-interferon (alpha-IFN) and ribavirin treatment of HCV hemodialysis (HDx) patients, by comparing the sustained virological response to that obtained by local historical data on treatment with alpha-IFN alone. A secondary aim was to establish the optimal therapeutic dose of ribavirin in this regimen. METHODS: Twenty HCV-HDx patients who were histologically (liver biopsy) and virologically (HCV-PCR)-positive were selected randomly. They received combination therapy with 3 million units (MU) of alpha-IFN and 200 mg of ribavirin three times a week. Initially nine patients were treated for 24 weeks. Later, another 11 patients were randomly selected to give the combination for 48 weeks. RESULTS: Six of the nine patients who were treated for 24 weeks (66%) became HCV-PCR-negative by the end of the treatment period. They continued to have a sustain virologic response at 6 months after the cessation of therapy. Six of the 11 patients (55%) who were treated for 48 weeks became HCV-PCR-negative at the end, and at 6 months after cessation of treatment. Of the first six responders, 4 (66%) maintained a sustained virologic response at 1 year postcessation of therapy. Nine of the 11 patients had genotype 4 and 1. No side effects were reported for a ribavirin dose of 200 mg three times a week. CONCLUSION: This pilot study suggests that combination treatment for 24 weeks and 48 weeks with 3 MU alpha-IFN and 200 mg ribavirin three times a week, elicited a sustained virologic response in HDx patients with HCV infection better than IFN alone with minimal side effects. A prospective, double-blind, controlled study using pegylated INF plus ribavirin is currently underway. PMID- 15350491 TI - Renal transplantation: seventeen years of follow-up in Qatar. AB - Among 432 patients receiving renal transplants (RT) between 1986 and 2002, 238 were Qatari nationals and 194, expatriates of mixed nationalities. Since 1986 when we started a local transplant program, 70 cases were performed at our center and 362 abroad. Diabetic nephropathy was the most common cause of end-stage renal disease among Qatar and chronic glomerulonephritis among expatriate patients. New onset diabetes was reported after the transplant operation in 7.3% of the cases. Recipient age ranged from 14 to 75 years with the mean of 48.5 years among diabetics and 34.5 years among nondiabetics. Acute rejection occurred in 19.2% with chronic allograft nephropathy in 16.2% of cases. Two-year survival rates at our center versus the abroad units were 98% and 97% for patients and 85.7% and 82.5% for grafts respectively. The mortality was mainly related to myocardial infarction, which occurred significantly more often among diabetics. Other causes of mortality, such as sepsis, hepatic failure, and cytomegalovirus infection, did not differ significantly between diabetic and nondiabetic patients. The donor source at our center was living related (78.6%), cadaver (18.5%), and living unrelated (2.9%) as compared to 29.3%, 6.6%, and 64.1% of those performed abroad, respectively. The 5-year survivals among living-unrelated allografts performed abroad was 45.2% compared to 64.3% in living-related and cadaveric donors. Despite the disappointing results, the existing shortage of local kidney donors persuades our patients to go abroad for living- unrelated transplants. Educational programs and incentives are recommended to increase the supply of cadaver organs. PMID- 15350492 TI - Breast fibroadenomas in renal transplant recipients. AB - Breast fibroadenomas are the most common solid lesions found in young women. We report on four patients who underwent renal transplantation and developed bilateral fibroadenomas while on cyclosporine (CsA). One patient developed symptomatic giant fibroadenomas and underwent bilateral mammoplasty. A significant decrease in the size of the breasts was noticed after switching to tacrolimus in three patients. Awareness of the association between CsA and fibroadenomas should help to achieve the correct diagnosis in transplant patients without subjecting them to unnecessary procedures. PMID- 15350493 TI - Late cytomegalovirus disease with atypical presentation in renal transplant patients: case reports. AB - Cytomegalovirus (CMV) disease typically occurs 1 to 4 months (median 35 days) after solid organ transplantation. Recent reports documented that the natural history of CMV disease associated with solid organ transplantation has been modified as a result of the widespread use of potent immunosuppressents and antiviral prophylaxis. We herein report three pretransplant CMV seropositive recipients (with unknown donor status) who were diagnosed recently to display late and atypical CMV disease. Two men and one woman included two patients who presented with allograft dysfunction at 12 years and at 3 years after transplantation. Both patients showed increased serum creatinine approximately from baseline 200 to >400 micromol/L over 3 months in the absence of features of rejection or cyclosporine toxicity. A renal biopsy was refused by both patients. Two of the three patients presented with symptoms of enterocolitis (diarrhea, nausea, weight loss), which had persisted for more than 6 months. Other symptoms and signs of overt CMV disease (fever, leukopenia) were absent. None had pulmonary, hepatic, or other major organ involvement. In all patients IgG antibodies and CMV DNA by polymerase chain reaction were positive with negative IgM antibodies. The immunosuppressive regimen consisted of mycophenolate mofetil (MMF), steroids, and calcineurin inhibitors. The kidney function significantly improved in both patients with renal dysfunction. Gastrointestinal symptoms resolved completely with gradual weight gain. The recognition and early diagnosis of late atypical CMV disease in kidney transplant patients presenting with allograft dysfunction and/or other organ systems is important. The MMF has a red herring effect in our cases due to its GI side effects. PMID- 15350494 TI - Distribution of HLA class II (DRB1/DQB1) alleles and haplotypes among Bahraini and Lebanese Arabs. AB - The genetic relationship between Bahraini and Lebanese Arabs in terms of HLA class II (DRB1 and DQB1) gene and haplotype frequencies was investigated in a group of 90 Lebanese and 52 Bahraini Arabs. Subjects of both sexes were unrelated and HLA-DRB1 and DQB1 genes were genotyped using the polymerase chain reaction sequence specific primer (PCR-SSP) technique. Analysis of the HLA-DRB1 alleles showed that the DRB1*040101 and DRB1*110101 alleles were more common among Lebanese, whereas DRB1*030101, DRB1*130701/1327, and DRB1*160101 alleles were more common among Bahrainis. Similarly, of the 7 HLA-DQB1 alleles analyzed, the presence of DQB1*0201 was higher among Bahrainis, whereas DQB1*030101 was higher among Lebanese. The DRB1*160101-DQB1*050101 (23.08%) and DRB1*030101-DQB1*0201 (21.15%) haplotypes were more frequent among Bahrainis, while the DRB1*110101 DQB1*030101 (56.67%), DRB1*040101-DQB1*0302 (28.89%) and DRB1*040101/DQB1*030101 (25.56%) haplotypes were more frequent in Lebanese subjects. Our results underline significant differences between these two populations in HLA class II distribution, and provide basic information for further studies of MHC heterogeneity among Arab-speaking countries, and as a reference for further anthropologic studies. PMID- 15350495 TI - Cytomegalovirus prophylaxis with ganciclovir in kidney transplant recipients receiving induction antilymphocyte antibodies. AB - BACKGROUND: Cytomegalovirus (CMV) is one of the serious viral infections after organ transplantation, especially in patients receiving anti-lymphocyte antibodies. Prevention of the infection using antiviral chemotherapy (ganciclovir) has gained interest in the transplant community due to the availability of quantitative methods for viral detection and monitoring. METHODS: Forty-six CMV seropositive kidney transplant recipients were assigned to receive induction immunosuppression with anti-thymocyte globulin (ATG, Fresenius). Prophylactic intravenous ganciclovir was administered for 2 weeks at a dose of 5 mg/kg/d (adjusted to kidney function) starting from the day of surgery. Patients were monitored regularly for CMV infection or disease over 1 year posttransplant. The time to CMV manifestation, the number of antigenemia assay-positive cells, the clinical severity of infection, the incidence of acute rejection, the graft function, and the duration of hospital stay were evaluated. This group was compared to a historical matched control cohort (n = 37) transplanted earlier who did not receive prophylactic ganciclovir. RESULT: The incidence of CMV disease was significantly less among the prophylaxis than the control group (6/46 patients [13%] vs 16/37 patients [43.2%], P = <.004). The time to develop CMV manifestations was much longer in the prophylaxis group than in the control group (median 92 vs 32 days, P /=20%. They received IVIG for a period of two weeks and a total of 6 doses. None of these patients, however, attained significant suppression of anti-HLA antibodies titer or a negative crossmatch reaction. We found that IVIG alone couldn't effectively inhibit preformed anti-HLA antibodies to allow successful renal transplantation. PMID- 15350497 TI - Larval damselflies in extreme environments: behavioral and physiological response to hypoxic stress. AB - The extensive papyrus (Cyperus papyrus) swamps of East and Central Africa form a habitat of great ecological importance due to their extent, the extreme and chronic hypoxia of the interior swamp, and the unique assemblages of water breathing insects that characterize these communities, including zygopteran (damselfly) larvae. The major goal of this study was to quantify physiological and behavioral responses of gilled and gill-less damselfly larvae of a papyrus swamp specialist, Proischnura subfurcatum, to low-oxygen conditions. Gill autotomization was common in P. subfurcatum of the Rwembaita Swamp in Kibale National Park, Uganda, with one to three gills missing from 56% of the specimens surveyed. We examined behavioral (ventilation activity and vertical migration) and physiological (metabolic rate) response to hypoxia in gilled and gill-less P. subfurcatum. Behavioral response to progressive hypoxia indicated that gill-less individuals rely more on use of wing sheaths (lifting and spreading) than gilled P. subfurcatum larvae. However, both morphs migrated to the surface to gain contact with atmospheric air under extreme hypoxia. On average, the rate of oxygen consumption of gill-less individuals was 51% lower than that of gilled individuals. This observed metabolic depression in gill-less P. subfurcatum may be attributed to the loss of major respiratory appendages. However, the apparent ability of both gilled and gill-less individuals to maintain their metabolic rates to a similar critical tension suggests other mechanisms may compensate for loss of gills, though not enough to mediate metabolic depression. PMID- 15350498 TI - Cytotoxic activity of blowfly Calliphora vicina hemocytes. AB - The cytotoxic activity of hemocytes isolated from larvae of the blowfly Calliphora vicina was tested using human myelogenous leukemia K562 cells as target. Both single cell and cytotoxicity assays demonstrated that the hemocytes recognize the K562 cells as nonself, firmly attach to their surface and induce target destruction in a manner resembling the effect of mammalian cytotoxic lymphocytes. The cytotoxic activity increased dramatically in the course of larval metamorphosis and was considerably higher shortly before the onset of pupariation, compared to the activity of human peripheral blood or mouse spleen lymphocytes. In insects, the cytotoxic hemocytes may take part in defense against eukaryotic parasites and in the elimination of aberrant self cells, as well as in developmental processes such as metamorphosis. PMID- 15350499 TI - Antioxidant defense of the midgut epithelium by the peritrophic envelope in caterpillars. AB - The peritrophic envelope (PE) is an extracellular matrix that is secreted by the midgut epithelium in most arthropods. In addition to protecting the midgut epithelium from abrasive food particles and microbial pathogens, in vitro experiments have suggested that the PE functions as a radical-scavenging antioxidant in caterpillars. This study tested the hypothesis that the PE is a "sacrificial antioxidant" in vivo in caterpillars. As a sacrificial antioxidant, the PE would (1) bind catalytic metal ions, (2) become oxidized itself, and (3) protect the midgut epithelium from oxidative damage. Each of these functions was supported by our results: the PE in Malacosoma disstria adsorbed increased amounts of iron as the concentration of iron was increased in its diet. Iron adsorption by the PE helped protect the midgut epithelium of M. disstria from oxidative damage over a wide range of ingested iron concentrations. Secondly, while the midgut epithelium was protected, protein oxidation in the PE increased 108% when tannic acid was oxidized in the endoperitrophic space. Finally, when the formation of the PE was inhibited by Calcofluor, protein carbonyls in the midgut epithelia of M. disstria and Orgyia leucostigma increased by two- to threefold. We conclude that the PE functions as an effective iron-binding and radical-scavenging antioxidant that protects the midgut epithelia of caterpillars. PMID- 15350500 TI - Patch-clamp study of the apical membrane of the midgut of Manduca sexta larvae: direct demonstration of endogenous channels and effect of a Bacillus thuringiensis toxin. AB - The patch-clamp technique was applied to the apical membrane of epithelial midgut cells of a lepidoptera, Manduca sexta L. Access to the apical membrane, the main target site of Bacillus thuringiensis (Bt) toxins, was achieved by using freshly isolated larval midgut preparations mounted onto holding glass pipettes. The epithelial cells retained their functional integrity, as evidenced by the magnitude of intracellular potentials recorded with microelectrodes. With standard 32 mM K(+) solution in the bath and the patch-clamp pipette, endogenous channel activity was detected in about 50% of experiments, mainly in moulting larvae and larvae that had been kept at reduced temperature for at least two days prior to the experiments. In both cell-attached and inside-out patch-clamp configurations, different types of channel were observed, with conductances varying between about 5 and 50 pS and different conducting properties. Addition of trypsin-activated Cry1Ac Bt toxin in the patch-clamp pipette triggered, after a delay, large conductances of a few nanosiemens. This is the first study allowing exploration, in the intact midgut, of the properties of apical membrane channels and the direct interaction between the apical membrane of epithelial cells and pathogenic agents such as Bt toxins. PMID- 15350501 TI - Differences between larval and pupal hemocytes of the tobacco hornworm, Manduca sexta, determined by monoclonal antibodies and density centrifugation. AB - Insect hemocytes play a major role in developmental processes where they disassociate and rebuild metamorphosing tissues while undergoing physiological changes themselves. We identified hemocyte changes from the last larval to the beginning of the pupal stage of the tobacco hornworm, Manduca sexta. Larval and pupal hemocytes behaved differently in a 40% Percoll density gradient. Larval granular cells were found in almost all density layers, pupal granular cells were abundant in high density layers; larval plasmatocytes occurred in dense layers, pupal plasmatocytes became enriched in less dense layers of the gradient. Using a panel of monoclonal antibodies generated against purified hemocytes, several different antibody binding patterns were identified. Quantitative differences in staining intensities were observed more often than qualitative changes, e.g. a loss or a gain of staining. Both phenomena were related to both plasmatocytes and granular cells. The distribution of the corresponding antigens in tissues was tested on cross sections of larvae and pupae as well as in Western blot analyses using organ homogenates. Several antibodies were specific for hemocytes only, among which two antibodies bound to molecules of the hematopoietic organ. Other antibodies had an additional reactivity to other tissues, mainly to the basal lamina. PMID- 15350502 TI - The effects of dopamine receptor agonists and antagonists on the secretory rate of cockroach (Periplaneta americana) salivary glands. AB - The acinar salivary glands of the cockroach, Periplaneta americana, are innervated by dopaminergic and serotonergic nerve fibers. Serotonin stimulates the secretion of protein-rich saliva, whereas dopamine causes the production of protein-free saliva. This suggests that dopamine acts selectively on ion transporting peripheral cells within the acini and the duct cells, and that serotonin acts on the protein-producing central cells of the acini. We have investigated the pharmacology of the dopamine-induced secretory activity of the salivary gland of Periplaneta americana by testing several dopamine receptor agonists and antagonists. The effects of dopamine can be mimicked by the non selective dopamine receptor agonist 6,7-ADTN and, less effectively, by the vertebrate D1 receptor-selective agonist chloro-APB. The vertebrate D1 receptor selective agonist SKF 38393 and vertebrate D2 receptor-selective agonist R(-) TNPA were ineffective. R(+)-Lisuride induces a secretory response with a slower onset and a lower maximal response compared with dopamine-induced secretion. However, lisuride-stimulated glands continue secreting saliva, even after lisuride-washout. Dopamine-induced secretions can be blocked by the vertebrate dopamine receptor antagonists cis(Z)-flupenthixol, chlorpromazine, and S(+) butaclamol. Our pharmacological data do not unequivocally indicate whether the dopamine receptors on the Periplaneta salivary glands belong to the D1 or D2 subfamily of dopamine receptors, but we can confirm that the pharmacology of invertebrate dopamine receptors is remarkably different from that of their vertebrate counterparts. PMID- 15350503 TI - Ecdysone-induced accumulation of mosquito cells in the G1 phase of the cell cycle. AB - We have established baseline conditions for investigating the interaction of the insect steroid hormone 20-hydroxyecdysone (20E) with the cell cycle in the C7-10 cell line from the mosquito, Aedes albopictus. As is the case with Drosophila melanogaster cells, treatment of C7-10 cells with 20E inhibits proliferation. In the presence of 10(-6) M 20E, a gradual decline in cell number is typically apparent at 24 h. Media components such as phenol red and the potential presence of endogenous steroids in serum have no effect on the response to 20E. Pre treating the cells with 10(-8) M 20E, with or without an intervening hormone-free period, did not alter the response to 10(-6) M 20E. However, replenishment of the medium appeared to synchronize the response to 10(-6) M 20E, causing an abrupt and complete cessation of cell division by 48 h. Flow cytometry over a 20 h period showed a decrease in the proportion of cells in S within 4-6 h after exposure to 20E. By 6-10 h, a transient increase in G2 was followed by the accumulation of more than 70% of the cells in G1. These data suggest that after treatment with 20E, cells complete the ongoing cycle before arresting in G1. Consistent with the decrease in the proportion of cells in S and G2, western blots show that levels of cyclin A, which is required during the S phase of the cycle, decreased in 20E-treated cells. PMID- 15350504 TI - The development of the endoparasitoid wasp Venturia canescens in superparasitised Ephestia kuehniella. AB - Using a molecular marker that allows the differentiation of two strains of the solitary endoparasitoid wasp Venturia canescens, the study investigated the influence of host mass and the time interval between ovipositions on the survival and development of larvae from both the first and second laid eggs in superparasitised Ephestia kuehniella. As the time interval between ovipositions increased both overall and superparasitism success decreased, however, time between, and order of, ovipositions had little effect on other developmental parameters. Adult size increased with host mass under both parasitism and superparasitism, while host mortality decreased with host mass under superparasitism. In addition, wasps emerging from superparasitised hosts were larger than wasps from parasitised hosts. The results confirm that for V. canescens on the host E. kuehniella both self- and conspecific-superparasitism will be an adaptive strategy when hosts are the limiting factor. PMID- 15350505 TI - The function of a trypsin-like enzyme in the saliva of Euplectrus separatae larvae. AB - Larvae of the gregarious ectoparasitoid, Euplectrus separatae, a species that parasitizes Pseudaletia separata, migrate from the dorsal to the ventral side of the host larva for pupation 7 days after parasitization. The parasitized host larvae die after the migration. The body mass of the parasitoid larvae increases while that of the host larva drastically decreases. Most of the tissue in the dead host larvae completely collapses. In this study, we examined the cause of host death and how the tissues collapse. Artificial removal of all parasitoid larvae before their migration on day 7 rescued the host larvae, but removal after parasitoid migration did not rescue the hosts. Tissues of the dead host larvae were completely liquefied. Injection of saliva from day 7 parasitoid larvae into host larvae killed the host larvae. High activity of a trypsin-like enzyme was detected in the saliva of day 7 parasitoids. Though phospholipase B and hyaluronidase were also detected in the saliva, commercial phospholipase B and hyaluronidase did not kill the hosts, whereas an injection of commercial trypsin was lethal. The trypsin-injected hosts showed the same tissue collapse as noted in parasitized and saliva-injected hosts. Leupeptin, a trypsin inhibitor, reduced mortality when injected into day 7 hosts (parasitoids were removal following migration). These observations suggest that the day 7 parasitoid larvae release saliva containing a trypsin-like enzyme to digest the host tissues following migration. PMID- 15350506 TI - Hormonal control of phase-related changes in the number of antennal sensilla in the desert locust, Schistocerca gregaria: possible involvement of [His7] corazonin. AB - The effect of [His(7)]-corazonin on the abundance of antennal sensilla in the desert locust, Schistocerca gregaria, was investigated to test the hypothesis that injection of this neuropeptide would mimic a crowding effect. Solitarious locusts (reared in isolation) were injected with [His(7)]-corazonin at the 3rd nymphal instar and the numbers of sensilla on the 2nd, 8th and 14th antennal segments in the adult stage were compared with those for oil-injected solitarious controls or un-injected gregarious locusts (reared in group). The numbers of sensilla on these antennal segments were all reduced significantly after [His(7)] corazonin injection compared with those for oil-injected controls, but similar to the values for gregarious individuals. Among the four major types of olfactory sensilla, coeloconic, trichoid, basiconic type A and basiconic type B, [His(7)] corazonin injection influenced the abundance of all but the last type. The effect of [His(7)]-corazonin injection varied with the time of injection; the earlier the injection the larger the effects on the abundance of total antennal sensilla on the 8th segment, although the way in which the injection affected the abundance varied with the sensillum type. A hypothesis explaining how crowding affects the abundance of antennal sensilla and other phase-related characteristics through changes in [His(7)]-corazonin concentrations was proposed. PMID- 15350507 TI - Optimum culture conditions for specific and nonspecific activation of whole blood and PBMC for intracellular cytokine assessment by flow cytometry. AB - The assessment of cytokine production is an important component of studies of cell-mediated immune responses (CMI) to immunological challenges. In this study, we present a method to enhance the detection of cytokine-producing cells by allowing antigen-specific cells to expand in long-term culture. We investigated the influence of the degree of dilution of whole blood and the duration of the incubation period on whole blood as well as peripheral blood mononuclear cells (PBMCs), cultured in the absence or presence of mitogens, superantigens or specific antigens, for intracellular cytokine production (IFNgamma, TNFalpha, IL 2, IL-4, IL-10 and IL-13) by CD4+ and CD8+ T lymphocytes using four-colour flow cytometry. Whole blood was diluted 1/1, 1/2, 1/5 and 1/10, and cultured for 6, 24, 48, 72 and 120 h in the presence of antibodies against the co-stimulatory molecules CD28 and CD49d, and, during the last 4 h of culture, in the presence of brefeldin A. Optimum conditions for detection of a high number of IFNgamma positive cells were observed after 72 h of culture in blood diluted 1/10. Median frequencies of IFNgamma+ cells obtained after activation by PMA-ionomycin, PHA or SEA-B were 29.3%, 20.0% and 6.8% for CD4+ cells, and 67.8%, 20.6% and 6.8% for CD8+ cells. In blood samples diluted 1/5 or 1/10, and cultured in the presence of cytomegalovirus (CMV) or varicella-zoster virus (VZV), mean peak levels of 2.8% and 1.4% IFNgamma+CD4+ cells were recorded at 120 h. The levels of cells producing cytokines other than IFNgamma were generally much lower and, in the case of IL-4 and IL-13, difficult to distinguish from background levels recorded in cultures with medium only. Kinetic studies of cytokine production by PBMCs showed a pattern similar to that of whole blood with peak levels of IFNgamma producing cells recorded at 72 h. The increased levels of IFNgamma production after culture for 72 h were due to an expansion of the numbers of cytokine producing cells responsive to a specific stimulus. Antigen-specific cells are usually present only at low levels in peripheral blood and may not be detected following simple activation for a few hours. To reach a level of detection in such cases, culture of diluted blood for several days is recommended. PMID- 15350508 TI - An alternative immunohistochemical method for detecting Leishmania amastigotes in paraffin-embedded canine tissues. AB - Canine visceral leishmaniasis (CVL) is a zoonosis and a chronic systemic disease of the dog caused by a protozoan by the species Leishmania infantum in the Old World and Leishmania chagasi in the New World. Several methods are currently employed for the diagnosis of CVL including microscopic detection of the parasite in bone marrow and lymph node aspirates, demonstration of specific antibodies anti-Leishmania in sera from infected animals, and isolation of the parasite by in vitro culture or by inoculation of laboratory animals. However, a definitive diagnosis is based on the actual detection of the parasite, which is conventionally achieved by examining Giemsa-stained smears or histopathological sections stained with hematoxylin and eosin. These methods have a low sensitivity, and therefore, they are often inconclusive. This is particularly true in canine organs that have a low level of parasitism such as kidneys, lungs, central nervous system, and testis, or, in some cases, the skin. The technique for immunohistochemical detection of leishmanial amastigotes in canine tissues has been reported previously and has proved to be undoubtedly efficient for the diagnosis. In this paper, we describe a straightforward and inexpensive immunohistochemical approach for Leishmania detection in formalin-fixed paraffin embedded canine tissues. Amastigote forms of Leishmania were easily observed within macrophages in several organs from naturally infected dogs using the streptavidin-biotin immunohistochemical method with canine hyperimmune serum as the primary antibody. In addition, the secondary antibody used was not specific to canine immunoglobulin, characterizing a cross-immune reaction. Our results indicate that this technique could be a useful tool for epidemiological, clinical, and histopathological studies. PMID- 15350509 TI - Optimal timing to repopulation of resident alveolar macrophages with donor cells following total body irradiation and bone marrow transplantation in mice. AB - To determine the time required to repopulate mouse lungs with donor alveolar macrophages following total body irradiation (TBI) and bone marrow transplantation (BMT), C57Bl/6 mice were subjected to TBI with 900 cGy, followed by transplantation of bone marrow cells from mice expressing green fluorescent protein (GFP) in their somatic cells. The mice were euthanized at either 30 (n=5), 60 (n=5) or 90 (n=5) days following BMT. Thirty days following transplantation, 87.8 +/- 3.9% (mean +/- S.E.M.) circulating leukocytes in recipient mice were derived from the donor, as determined by fluorescence activated cell sorting (FACS) analysis for GFP. However, only 46.9 +/- 7.4% of the resident alveolar cells expressed GFP, indicating incomplete repopulation. By day 60 post-transplantation, the percentage of bronchoalveolar lavage fluid (BALF) cells expressing GFP reached 74.5 +/- 2.4%, remaining stable 90 days after transplantation (80.4 +/- 1.9%). We conclude that 60 days after TBI with 900 cGy and bone marrow transplantation, the majority of the lung resident alveolar macrophages is of donor origin. This study provides useful information regarding the time of reconstitution with donor alveolar macrophages in the pulmonary airspaces of recipient mice following marrow transplantation. PMID- 15350510 TI - Monitoring an antibody affinity chromatography with a label-free optical biosensor technique. AB - In the production of polyclonal antibody, a purification step is necessary which is often done by affinity chromatography. We present a biosensor system based on reflectometric interference spectroscopy (RIfS) to monitor the quantity and quality in terms of affinity and kinetic constants of the antibody during this procedure. Biosensors are rapid compared with ELISA, which is done in practice and can work fully automated. They provide additional information about the active antibody to protein concentration ratio and the affinity of the antibody. We show how to determine these values very accurate. In addition, we describe a new rapid method to monitor the affinity chromatography in process. This gives the possibility to select antibody fractions with best properties in respect to the application. PMID- 15350511 TI - Use of serial analysis of gene expression (SAGE) technology reveals new granulocytic markers. AB - Because of its short life span in blood, its low RNA content and its highly condensed nucleus, the granulocyte was initially considered as a terminally differentiated cell unable to express novel genes. However, mature granulocytes still contain a variety of mRNAs and may respond to external stimuli by rapid and complex changes in gene expression. The present work was undertaken to provide a wider view of the gene expression profile in unstimulated circulating PMNs. We used serial analysis of gene expression (SAGE) adapted to downsized extracts (SADE) to cope with their small mRNA content. As cell samples of the highest degree of purity were crucial for this project, we adapted a magnetic cell sorting method to reach the required high levels of purity (99.55 +/- 0.19%) together with low activation rates (1.37 +/- 0.28). We analyzed 20,787 tags identifying 8547 different transcripts, of which 47.8% were unknown, 34.6% were transcripts of known genes, and 13.8% matched with expressed sequence tags (EST). Highly expressed genes were those involved in cell mobility, diapedesis, cell signaling and destruction of micro-organisms. In addition, this method led to the identification of genes which had not previously been reported in granulocytes. These results could provide new molecular markers and a reliable reference for the investigation of pathologies involving alteration of the granulocyte gene expression profile. PMID- 15350512 TI - Identification of compounds that enhance the anti-lymphoma activity of rituximab using flow cytometric high-content screening. AB - In this report, we describe a new flow cytometry technique termed flow cytometric high-content screening (FC-HCS) which involves semi-automated processing and analysis of multiparameter flow cytometry samples. As a first test of the FC-HCS technique, we used it to screen a 2000-compound library, called the National Cancer Institute (NCI) Diversity Set, to identify agents that would enhance the anti-lymphoma activity of the therapeutic monoclonal antibody rituximab. FC-HCS identified 15 compounds from the Diversity Set that significantly enhanced the ability of rituximab to inhibit cell cycle progression and induce apoptosis in lymphoma cells. The validity of the screening results was confirmed for several compounds using additional assays of cell proliferation, apoptosis and cell growth. The FC-HCS technique was relatively simple and reliable and could process up to 1000 samples/day on a single flow cytometer. The FC-HCS technique may be useful for a variety of applications including drug discovery, immunologic monitoring of patients, functional genomics studies and tissue engineering efforts. PMID- 15350513 TI - Isolation and tracking of a rare lymphoid progenitor cell which facilitates bone marrow transplantation in mice. AB - Bone marrow cells are composed of pluripotent stem cells to terminally differentiated cells, with a wide variety of abundance of each cell type. In the past, many of the cell types within this heterogeneous population have been characterized either by expression of specific proteins or using functional markers. In spite of promising results obtained with the latter method, various cell types within bone marrow have not been well characterized due to the low abundance of a specific cell type. Considering the demand for a reliable technique to enrich cell types, a wide variety of approaches, ranging from simple nylon wool columns to high-speed cell sorting, have evolved. Only limited success has been obtained with approaches ranging from the detection of MHC antigen to positron emission tomography to track the ontogeny of specific bone marrow derived cells in studies of syngeneic or allogeneic transplantation. The present study describes a relatively simple method to enrich and track a rare bone marrow cell (facilitating cell, FC), which can facilitate allogeneic bone marrow stem cell transplantation in mice. The isolation technique is comprised of enrichment of FC by magnetic activated cell sorting (MACS) system followed by purification through high-speed cell sorter. An initial inoculation of 30,000 FC obtained from male mice was detected in the thymus, spleen, and bone marrow of allogeneic female recipients, by using 32P-labeled dCTP in a specific PCR for Y-chromosome. This technique may improve the efficiency of isolation of other rare cells from the bone marrow. PMID- 15350514 TI - Definition of the fine specificity of the monoclonal antibody 81D4: its reactivity with lysine and polyamine isopeptide cross-links. AB - The 81D1C2 monoclonal antibody (Mab) directed against the Nepsilon-(gamma-L glutamyl)-L-lysine isopeptide was found to cross-react on Enzyme Immuno Assay (EIA) with acylated lysines. Using a differential screening EIA procedure, a new Mab 81D4 was selected, which did not cross-react with acylated lysines but exhibited strong reactivity with Nepsilon-(gamma-L-glutamyl)-L-lysine formed by covalently coupling the gamma-carboxyl of NalphaCBZ OtBu glutamic acid to epsilon NH2 derivatized microtiter plates. When Nepsilon-(gamma-L-glutamyl)-L-lysine isopeptides were generated on gamma-carboxyl derivatized plates, only lysine isopeptides with blocked alpha-amines were reactive, regardless of whether the bond formed by the amine blocking agent was a carbamate with carbobenzyloxychloride or an amide with acetic anhydride. The 81D4 Mab showed little or no affinity for free Nepsilon-(gamma-L-glutamyl)-L-lysine (IC50>5 mM), for N1 or N4 mono(gamma-Poly L-glutamyl)putrescine, and for N1 mono(gamma-Poly L glutamyl)spermidine (IC50>5 mM). However, when these same isopeptides were synthesized as cross-links between two protein chains--Nepsilon-(gamma-L glutamyl)-L-lysine between Poly L-glutamate and Poly L-lysine; N1N4 -bis(gamma Poly L-glutamyl)putrescine, N1N8 -bis(gamma-Poly L-glutamyl)spermidine between Poly-L-glutamate chains--very good reactivity was observed (IC50 400 microM for lysine; 80 microM for putrescine and spermidine). In addition to the chemically synthesized isopeptide cross-links that were recognized by this Mab, the naturally occurring Nepsilon-(a-L-glutamyl)-L-lysine isopeptide cross-links in D dimer, which are formed by the action of plasma transglutaminase (Factor XIII) on fibrin, were also detected on immunoblots using 81D4 as the primary antibody. PMID- 15350515 TI - Immunodetection and quantification of cytochromes P450 using epitope tagging: immunological, spectroscopic, and kinetic analysis of cinnamate 4-hydroxylase. AB - Cytochrome P450-dependent monooxygenases (P450s) are integral membrane proteins typically expressed at low levels both in vivo and by heterologous expression systems, often making quantification of these enzymes challenging. Since the time of their discovery, P450s have typically been quantified by their carbon monoxide (CO) difference spectra. Although this technique is reliable, it requires quantities of enzyme that are sometimes difficult to obtain, and spectroscopic instruments and expertise frequently unavailable in laboratories whose primary focus is genetics or molecular biology. We have developed a method for quantifying recombinant FLAG epitope-tagged proteins using fluorescence detection of a chromophore-labeled anti-FLAG monoclonal antibody and well-established immunoblot technology. The utility of this technique was tested using cinnamate 4 hydroxylase (C4H), one of the best-studied plant P450s. No substantial differences in the stability or kinetic properties were observed between the native and FLAG-tagged enzymes. Immunochemical quantification of epitope-tagged C4H reported slightly lower P450 concentrations than conventional methods but has a limit of quantification 400-fold lower than carbon monoxide difference spectroscopy. PMID- 15350516 TI - Establishment of an ELISA system for determining soluble LAIR-1 levels in sera of patients with HFRS and kidney transplant. AB - LAIR-1, the leukocyte-associated Ig-like receptor-1, is a trans-membrane molecule that functions as an inhibitory receptor on natural killer cells, T lymphocytes and monocytes. It has been well known that many trans-membrane receptors can shed from the cell surface and be released into the circulation in soluble form when lymphocytes, endothelials and other immune cells are activated. In many cases, the levels of soluble receptors in the circulation can be used as markers of lymphocyte activation in transplant patients and virus infection patients. To investigate whether LAIR-1 is able to be released into the sera, we developed a sandwich enzyme-linked immunosorbent assay (ELISA) system based on two anti-LAIR 1 monoclonal antibodies (MAb) with different epitope specificities. Using this ELISA, we found that sLAIR-1 existed in the supernatants collected from PMA, PHA or CD3 MAb-stimulated lymphocytes cultures in vitro for the first time. Moreover, we found that LAIR-1 level in serum samples from healthy individuals was 6.2 +/- 3.3 ng/ml, whereas the levels in sera of patients with hemorrhagic fever with renal syndrome (HFRS) and patients 3-7 days after kidney transplant increased to 47.2 +/- 35.9 and 24.4 +/- 16.0 ng/ml, respectively. Furthermore, HFRS patients in oliguric phase showed higher serum sLAIR-1 levels than those in other phases, and transplant patients with rejection showed higher serum sLAIR-1 level than those without rejection. These findings demonstrated that LAIR-1 can be released when lymphocytes are activated, suggesting sLAIR-1 may be used as a predictor for monitoring immune reaction in some virus infections and organ transplants which may be useful in clinical treatment of these diseases. PMID- 15350517 TI - Development of a respiratory burst assay using zebrafish kidneys and embryos. AB - The innate immune response constitutes the first line of defense against invading pathogens and consists of a variety of immune defense mechanisms including the respiratory burst of phagocytes. Respiratory burst can be used as a reliable measure of the immune response of a host, and numerous assays have been developed to measure this response in a variety of mammal and fish species. Phagocytes, like granulocytes and macrophages, that are derived from different tissues, or grown in cell culture, have been employed in a range of assay formats employing a variety of detection methods. The small size of the zebrafish has prevented the large-scale extraction of these cells for respiratory burst assays in the zebrafish. In this work, we describe a respiratory burst assay developed for the zebrafish using intact kidneys and embryos as sources of phagocytes. Phorbol myristate acetate (PMA)-inducible reactive oxygen species (ROS) were detected following the oxidation of a non-fluorescent dye 2',7'-dihydrodichlorofluorescein diacetate (H2DCFDA) to dichlorofluorescein (DCF), a fluorescent product. Embryos from 1 day post-fertilization until 5 days post-fertilization (dpf) were employed in this assay. Abrogation of H2DCFDA oxidation by the protein kinase C (PKC) inhibitor bisindolylmaleimide I (BisI) indicated a reduction in the respiratory burst. Fluorescence from the PMA-induced respiratory burst in kidneys and embryos was significantly elevated above DMSO-treated controls, while preincubation with BisI inhibited the increase in fluorescence. Colocalization of cell-associated chloromethyl-dihydrodichlorofluorescein diacetate (CM-H2DCFDA) with the phagocyte selective dye neutral red is consistent with the observation that macrophages and granulocytes are the ROS-producing cells in the zebrafish. PMID- 15350518 TI - Antibody zymography: a novel adaptation of zymography to determine the protease neutralising potential of specific antibodies and snake antivenoms. AB - A common problem in the development of antibody-based therapeutics is the selection, usually from a large population, of specific antibodies with the desired function. One of our research objectives is to identify antibodies capable of neutralising the most important haemorrhagic and haemostasis disruptive proteases from viper venom. Here, we describe a modification of conventional gelatin-zymography that permits the identification of antibodies capable of neutralising gelatinolytic proteases. We demonstrate that the gelatinolytic activity of viper venom proteases is neutralised by addition of viper antivenom to the matrix of conventional gelatin-zymograms. Venom protein gelatinolytic activity was unaffected by inclusion of antibody from control, non immunised animals or immunoglobulin-depleted serum. The application of this antibody zymogram technique for future research on snake venoms is evaluated in the context of identified limitations. PMID- 15350519 TI - Rapid grouping of monoclonal antibodies based on their topographical epitopes by a label-free competitive immunoassay. AB - Topography of epitopes of monoclonal antibodies (MAbs), identified as the mutual competition of the MAbs, can be valuable indicators for the biological functions of MAbs. However, the determination of topographical epitopes is not performed before the functional screening of MAbs, because the requirement for purifying and labeling of MAbs makes the mapping experiment difficult, particularly in the early stage of MAb production. Here we describe a new label-free competitive enzyme-linked immunosorbent assay (LFC-ELISA) for the rapid grouping of MAbs based on the topography of their epitopes. In the LFC-ELISA, the immune complex formed by a competitor, MAb#2, and an antigen is challenged by an indicator, MAb#1 that had been captured on the ELISA plate through a secondary antibody. The MAb#2-antigen immune complex is trapped by MAb#1 only if MAb#1 reacts with an epitope different from that of MAb#2. The immune complex (MAb#2-antigen-MAb#1) is detected with an enzyme-labeled reagent specific to a tag on the antigen. Our experiments using different anti-CD30 MAbs and a CD30-Fc fusion protein as the antigen revealed that the LFC-ELISA performed well with MAbs of different isotypes (IgG1, IgG2a, and IgG2b), and in a practical range of MAb concentrations (0.3-10 microg/ml) and affinities (0.9-13 nM of Kd). We obtained pairwise competition data from all 26 anti-CD30 MAbs. We then utilized a cluster analysis and a bootstrap method to analyze the competition data for grouping of the MAbs. This objective and automated analysis identified eight distinct topographical epitopes on CD30. The reactivity of the anti-CD30 MAbs in immunoblot, and their inhibiting activity on CD30-CD30-ligand binding correlated with the topographical epitopes. The results show that the LFC-ELISA combined with cluster analysis is a useful new method for grouping MAbs based on their topographical epitopes and can be used in the early stage of MAb production. One useful application is to identify MAbs reacting with different epitopes from a large number of MAbs so that the most appropriate MAbs can be selected for therapeutic use. PMID- 15350520 TI - Quantitative Plasmodium sporozoite neutralization assay (TSNA). AB - The circumsporozoite (CS) protein is the major surface protein of Plasmodium sporozoites. Antibodies to the immunodominant repeat domain of CS immobilize sporozoites and prevent infection of hepatocytes. Plasmodium falciparum vaccines containing CS repeats are undergoing human trials in endemic areas, and proof of efficacy has been obtained. The correlates of protection are under investigation. Levels of anti-repeat antibodies in the serum of the human volunteers have been measured mostly by enzyme-linked immunosorbent assay (ELISA) and IFA. Assays that measure the effect of the serum antibodies on parasite infectivity (serum neutralization assays SNAs) are not usually performed because they require a susceptible host and P. falciparum sporozoites are highly infectious only to humans. To overcome this limitation, we developed a new assay named transgenic sporozoite neutralization assay (TSNA) that uses as neutralization target, a transgenic rodent malaria parasite Plasmodium berghei that bears the P. falciparum CS repeats [CS(Pf)]. Following incubation with human serum, CS(Pf) infectivity of HepG2 cells is evaluated by real-time PCR. We have compared ELISA titers and TSNAs in a limited number of sera from humans immunized with (T1B)4 MAP, a peptide vaccine containing P. falciparum CS repeats. A comparison between the two assays did not reach significance (p=0.175) when analyzed by non parametric Spearman correlation method. Ongoing human trials of CS-based vaccines should provide an opportunity to determine whether TSNAs will provide better correlates of protective immunity than ELISA assays. PMID- 15350521 TI - A novel assay to quantify cell death after transient expression of apoptotic genes in B- and T-lymphocytes. AB - We developed an assay allowing the detection and quantification of cell death after transient expression of apoptotic genes in B- and T-lymphocytes. For efficient gene transfer, B- and T-cells were electroporated under optimized conditions. To blind out the high background of non-transfected cells and cell death caused by the electroporation procedure itself, the green fluorescent protein (GFP) was co-transfected with the gene of interest. However, if the gene of interest was a potent apoptosis inducer, most successfully transfected cells were killed before GFP was expressed to levels sufficient for standard flow cytometry analysis or apoptosis assays. After staining of the transfected cells with propidium iodide (PI), very few GFP+/PI+ cells were detectable. To overcome this problem, the cell death rate induced by the transiently expressed gene was determined as the reduction of living green cells in the apoptotic versus a reference sample. This was achieved by an advanced flow cytometrical analysis quantifying the number of surviving green cells in normalised sample volumes directly relating to the number of initially transfected cells. Functioning of the assay was demonstrated by transient transfection of the potent apoptosis inducers TNF-receptor-associated death domain protein (TRADD) and a fusion protein of the transmembrane domain of the latent membrane protein 1 (LMP1) of Epstein-Barr virus and the signaling domain of TNF-receptor 1. We successfully applied the assay to the Burkitt lymphoma cell line BJAB and the T-leukemia cell line Jurkat. PMID- 15350522 TI - A toll-like receptor-based two-hybrid assay for detecting protein--protein interactions on live eukaryotic cells. AB - Protein--protein interactions underly diverse biological processes. Here, we describe a method for detecting protein interactions on the cell surface. This method is based on the mechanism of TLR2 activation whereby extracellular (EC) domain-mediated heterodimerization of TLR2 and TLR1 activates NF-kappaB and other signalling processes. Test proteins were expressed as the EC domains of TLR1 and TLR2 in fusion with the transmembrane/cytoplasmic (TM/Cyt) domains, i.e. tmTIR1 and tmTIR2. The feasibility of this TIR1/2-based method was examined by expression of IL-4 and the EC domains of the interleukin-4 receptor alpha (IL 4Ralpha) and the cytokine receptor common gamma chain (gammaC) as hybrid receptors with tmTIR1 and tmTIR2. Upon co-expression of IL4Ralpha-TIR1 and gammaC TIR2 in 293T cells, NF-kappaB activation was found to be inducible by IL-4. Co expression of IL4-TIR1 with IL4Ralpha-TIR2, but not gammaC-TIR2, led to constitutive NF-kappaB activation. This is consistent with IL-4 primarily binding to IL4Ralpha but not gammaC. Co-expression of the IL4Ralpha-TIR1/2, IL4-TIR1/2 or gammaC-TIR1/2 hybrid receptor pairs also constitutively activated NF-kappaB suggesting that IL-4, IL4Ralpha and gammaC form homodimers or homotypic interactions. This was confirmed by immunoprecipitation studies. In summary, we report a TIR1/2-based assay for detecting interactions between membrane proteins, receptors/ligands and secreted proteins on live eukaryotic cells. PMID- 15350523 TI - High-throughput imaging of bacterial colonies grown on filter plates with application to serum bactericidal assays. AB - The ability to accurately enumerate viable bacteria has applications in antibiotic screening assays, toxicology testing, and serological assays for functional antibodies. An impediment to high-throughput bacterial assays is the requirement to grow bacteria as individual colonies on semisolid media containing agar. We have now developed a method for growth, staining, and counting of bacterial colonies in 96-well filter plates. A unique feature of the method is that colony size is inversely proportional to the number of colonies in each well, presumably due to nutrient depletion. As a result, as many as 300 colony forming units (cfu) can be detected as discrete colonies within a single assay well. The resulting colonies can be counted automatically using an imaging system originally developed for ELISPOT assays. The method has been applied to the measurement of serum bactericidal activity (SBA) in human sera. PMID- 15350524 TI - Label-free, real-time monitoring of IgE-mediated mast cell activation on microelectronic cell sensor arrays. AB - Immunoglobulin E (IgE)-mediated mast cell activation is involved in the immediate phase of allergic reactions and plays a central role in the onslaught and persistence of allergic diseases. IgE-mediated mast cell activation includes two important events: cell sensitization resulting from IgE binding to Fc (FcepsilonRI) receptor and cell activation triggered by allergen-mediated oligomerization of membrane-bound IgE. Real-time monitoring of these events is needed to dissect the molecular mechanisms underlying IgE-mediated mast cell activation. Existing technologies are limited to label-based end-point assay formats, which detect either early signaling or final phase of mast cell activation. We describe a microelectronic cell sensor-based technology allowing dynamic monitoring of IgE-mediated mast cell sensitization and activation in real time without any labeling steps. RBL-2H3 mast cells were cultured onto the surface of microelectronic cell sensor arrays integrated into the bottom of microtiter plates, which record electric properties, such as impedance between cell membrane and sensor surface. In the presence of the allergen, dinitrophenyl (DNP)-bovine serum albumin (BSA), anti-DNP IgE-sensitized cells were activated within 5 min and the entire activation process was quantitatively and continuously recorded. Impedance measurements correlate with morphological dynamics and mediator release as measured by beta-hexosaminidase activity, and can be blocked by pharmacological agents, inhibiting IgE-mediated signaling. The assay on microelectronic cell sensor arrays can be scaled up for high-throughput screening of pharmacological inhibitors of IgE-mediated mast cell activation and other cell-based receptor-ligand assays. PMID- 15350525 TI - A flow cytometric method for determining the binding of coagulation factor X to monocytes in whole human blood. AB - It is known that inflammation affects the coagulation pathway, but the mechanisms are not clear. Because a persistent inflammatory condition is associated with several chronic diseases, including cardiovascular disorders, there is intense interest in determining if and how chronic inflammation contributes to a hypercoagulable state. One pathway by which inflammation interacts with coagulation is via monocyte binding and activation of coagulation Factor X (FX). Upon activation, monocytes express the alphaMbeta2 integrin CD11b/CD18, which has a binding site for the plasma protein FX. Binding is followed by the cleavage of FX into its activated form Xa(FXa) which, in turn, is responsible for the conversion of prothrombin to thrombin. To assess the contribution of this pathway, a straightforward assay in whole blood is needed for studies of inflammation-induced coagulation and thrombosis. The current assay for FXa binding requires isolation of the monocytes and measurement of bound FXa activity with a chromogenic substrate. Harvesting a sufficient number of monocytes for analysis requires a relatively large blood sample. In addition, it is known that the process of isolating neutrophils and monocytes from whole blood induces an upregulation of CD11b. Thus, the measurement process itself causes an artifact in receptor expression resulting in an overestimate of true state of monocyte activation and FX binding. To address these limitations, we developed a flow cytometric assay to directly measure the binding of FX to monocytes in whole blood. In this report we describe the methods of the procedure in detail and apply the procedure to demonstrate a significant increase of both monocyte CD11b expression and FXa binding when human blood samples were activated with the endotoxin, lipopolysaccharide in-vitro. PMID- 15350526 TI - Cancer vaccine development: on the way to break immune tolerance to malignant cells. AB - Exploiting a naturally occurring defense system, the immunotherapeutic approach embodies an ideal nontoxic treatment for cancer. Despite the evidence that immune effectors can play a significant role in controlling tumor growth either in natural conditions or in response to therapeutic manipulation, the cascade of molecular events leading to tumor rejection by the immune system remains to be fully elucidated. Nevertheless, some recent tumor immunology advancements might drastically change the way to design the next generation of cancer vaccines, hopefully improving the effectiveness of this therapeutic approach. In the present work, we will focus on three main areas of particular interest for the development of novel vaccination strategies: (a) cellular or molecular mechanisms of immune tolerance to malignant cells; (b) synergism between innate and adaptive immune response; (c) tumor-immune system interactions within the tumor microenvironment. PMID- 15350527 TI - A paradoxical pro-apoptotic effect of thrombin on smooth muscle cells. AB - Whereas thrombin (below 10 nM) is a potent mitogen, recent studies report that exposure to higher doses of thrombin could lead to apoptosis of neurons and tumor cells. Our results show that prolonged exposure (> or = 24 h) to thrombin (50-100 nM) exerts a pro-apoptotic effect on cultured vascular smooth muscle cells (VSMCs). This phenomenon depends on thrombin serine-protease activity but is independent of PAR-1 and -4 activation and subsequent signaling. The parallel occurrence of cell retraction and cleavage of fibronectin suggests that thrombin induced apoptosis is consecutive to pericellular proteolysis. These data point to a new potential action of thrombin in the cardiovascular system. PMID- 15350528 TI - Angiogenesis gene expression profiling in xenograft models to study cellular interactions. AB - The present study describes a method to simultaneously obtain the angiogenic expression profile in tumor cells and vascular cells of a single tumor. Human- and mouse-specific primers were used for quantitative real-time RT-PCR to determine the expression of vascular endothelial growth factors A, B, C, and D, vascular endothelial growth factor receptors 1, 2, and 3, neuropilin 1 and 2, angiopoietin 1, 2, 3/4, tyrosine kinase receptors 1 and 2, basic fibroblast growth factor (bFGF) in xenograft tumors obtained by injection of human ovarian carcinoma cells in nude mice. In addition, the effect of treatment with anginex and taxol on the expression profile was analyzed. Most factors were expressed higher in vascular cells as compared to tumor cells. In response to treatment, tumor cells significantly upregulated bFGF expression and downregulated VEGF receptor expression. This was accompanied by downregulation of VEGF-B and -D, and upregulation of angiopoietin-3 as well as angiopoetin receptors in nontumor cells. In conclusion, real-time qRT-PCR combined with xenograft tumor models presents a sensitive method to monitor angiogenesis and to analyze interactions between tumor cells and nontumor cells in vivo. The approach can be applied to different research fields in which xenograft models are used. PMID- 15350529 TI - Regulated compartmentalization of the putative DEAD-box helicase MDDX28 within the mitochondria in COS-1 cells. AB - We recently cloned a putative DEAD-box helicase MDDX28 and found that it was localized to the nuclei and mitochondria of COS-1 cells. The mitochondrial localization of MDDX28 is largely diffuse. We have, however, used immunofluorescence and immunogold cytochemistry to show that the MDDX28 protein is localized in a distinct mitochondrial subcompartment in 5-10% of COS-1 cells. This proportion increases to approximately 35% after treatment with ethidium bromide, suggesting upregulation following transcription inhibition. To our knowledge, this is the first example of protein relocation in the mitochondria caused by transcription inhibition. The mitochondrial subcompartmentation of MDDX28 was negatively affected by mutations in a RNA-binding domain and three basic domains previously shown to be important in transcription-dependent intranuclear localization. Furthermore, immunogold cytochemistry and fractionation of rat liver indicated that the protein is a part of an RNA-protein (RNP) complex interacting peripherally with the mitochondrial inner membrane. Our results reveal new principles for regulation of protein localization in the mitochondria and suggest parallels between the function of the MDDX28 protein in the nucleus and mitochondria. PMID- 15350530 TI - The long myosin light chain kinase is differentially phosphorylated during interphase and mitosis. AB - We have shown previously that the activity of the long myosin light chain kinase (MLCK) is cell cycle regulated with a decrease in specific activity during mitosis that can be restored following treatment with alkaline phosphatase. To better understand the role and significance of phosphorylation in regulating MLCK function during mitosis, we examined the phosphorylation state of in vivo derived MLCK. Phosphoamino acid analysis and phosphopeptide mapping demonstrate that the long MLCK is differentially phosphorylated on serine residues during interphase and mitosis with the majority of the phosphorylation sites located within the N terminal IgG domain. Biochemical assays show that Aurora B binds and phosphorylates the IgG domain of the long MLCK. In addition, phosphopeptide maps of the endogenous full-length MLCK from mitotic cells and in vitro phosphorylated IgG domain demonstrate that Aurora B phosphorylates the same sites as those observed in vivo. Altogether, these studies suggest that the long MLCK may be a cellular target for Aurora B during mitosis. PMID- 15350531 TI - The epithelial-mesenchymal transition of colon carcinoma involves expression of IL-8 and CXCR-1-mediated chemotaxis. AB - The epithelial-mesenchymal transition (EMT) is an essential component of embryonic development, tissue remodeling, and wound repair. In addition, many epithelial tumors, including colorectal carcinomas, appear to undergo this transition that may facilitate their invasion. Using a novel model of EMT in colon carcinoma in which the inflammatory cytokine TNF-alpha accelerates this TGF beta directed process, we report that TNF-alpha stimulation upregulates expression of the chemokine IL-8, and that this upregulation is dependent on the transcription factor NF-kappaB. Significantly, this effect is not merely an inflammatory response by these colon carcinoma cells because IL-8 expression is induced in cells undergoing a TGF-beta-driven EMT in the absence of exogenous TNF alpha. During the EMT, a concomitant increase in the chemokine receptor CXCR-1, but not CXCR-2, also occurs. Moreover, both IL-8 and CXCR-1 function in the chemokinetic and chemotactic migration of colon carcinoma cells as assessed by antibody inhibition studies. These studies establish that the regulated expression of a specific chemokine and its receptor are linked to the EMT and they provide a biochemical framework for understanding the mechanisms by which the EMT promotes migration. PMID- 15350532 TI - BMP-2 exerts differential effects on differentiation of rabbit bone marrow stromal cells grown in two-dimensional and three-dimensional systems and is required for in vitro bone formation in a PLGA scaffold. AB - Osteogenic differentiation of bone marrow stromal cells (BMSC) in a three dimensional (3-D) scaffold has not been well studied. In this work, we studied expression of bone-related genes during differentiation of rabbit BMSCs in response to bone morphogenetic protein (BMP)-2 in both 2-D and 3-D culture systems. When BMSCs were cultured on films (2-D) of biodegradable poly(lactide-co glycolide) (PLGA), increases in mRNA expression of type I collagen (Col I) and vascular endothelial growth factor (VEGF) became evident after 1 week. However, expression of both genes was only mildly stimulated by BMP-2. Expression of the osteopontin gene was highly stimulated by BMP-2 treatment. Expression of chordin, a BMP antagonist, increased significantly after 7 days. The increase was abrogated by BMP-2 treatment. BMP-2 was also able to stimulate mineralization of cultured BMSCs. After cells were switched to 3-D PLGA scaffolds for 24 h, expression of osteopontin and VEGF were markedly increased while expression of type I collagen and chordin remained unchanged. Expression of Col I did not increase with time in a 3-D culture as it did when cells were cultured on a 2-D film. We further explored the possibility of engineering bone tissue in vitro by seeding BMSCs into PLGA scaffolds. Cellular differentiation and bone formation in the scaffolds were analyzed histologically at 2 weeks and 2 months. Secretion of ECM by cells was evident at both 2-week and 2-month scaffolds, and was enhanced by rhBMP-2. Striking differences in 2-month scaffolds were observed between BMP treated and untreated cells. A woven bone-like structure appeared in the scaffolds treated with BMP-2. The structure was verified to be bone-related by: (1) the presence of organized collagen fibrils; (2) the presence of mineral; and (3) morphological features of trabecular bone. Although collagen was abundant in the untreated 2-month scaffolds, it was disorganized. The untreated scaffolds also lacked mineral deposits, which were present in 2-D cultured cells even in the absence of BMP-2. Our results indicate that the requirement of osteo inductive agents, such as BMP-2, is essential for bone tissue engineering. PMID- 15350533 TI - Structural changes in human type I collagen fibrils investigated by force spectroscopy. AB - In the field of biomechanics, collagen fibrils are believed to be robust mechanical structures characterized by a low extensibility. Until very recently, information on the mechanical properties of collagen fibrils could only be derived from ensemble measurements performed on complete tissues such as bone, skin, and tendon. Here, we measure force-elongation/relaxation profiles of single collagen fibrils using atomic force microscopy (AFM)-based force spectroscopy (FS). The elongation profiles show that in vitro-assembled human type I collagen fibrils are characterized by a large extensibility. Numerous discontinuities and a plateau in the force profile indicate major reorganization occurring within the fibrils in the 1.5- to 4.5-nN range. Our study demonstrates that newly assembled collagen fibrils are robust structures with a significant reserve of elasticity that could play a determinant role in the extracellular matrix (ECM) remodeling associated with tissue growth and morphogenesis. PMID- 15350534 TI - Identification and molecular characterization of Mnk1b, a splice variant of human MAP kinase-interacting kinase Mnk1. AB - In this paper, we report the identification and molecular characterization of a splice variant of human Mnk1 which has been named as Mnk1b. Human Mnk1b mRNA is homologous to human Mnk1 mRNA but lacking a region corresponding to exon 19, which causes a change in the reading frame generating a stop codon. The resulting protein lacks the last 89 amino acids at the C-terminal region that are replaced by 12 amino acids with an entirely new sequence. The C-terminal end in Mnk1 corresponds to the extracellular signal-regulated kinase (ERK1/2) binding site. Although Mnk1b lacks this domain and, consequently, is not phosphorylated by ERK1/2, it is able, however, to phosphorylate eIF4E in vitro and in vivo in a mitogen-activated protein kinases-independent manner. This result suggests that Mnk1b may play a key role in regulating protein translation in the absence of stimuli. Interestingly, a significant population of cells shows Mnk1b within the nucleus whereas Mnk1 is always detected in the cytoplasm. This fact may be explained because Mnk1b maintains the nuclear localization signal (NLS) but lacks the nuclear export sequence (NES). PMID- 15350535 TI - Wrch1 is a GTPase-deficient Cdc42-like protein with unusual binding characteristics and cellular effects. AB - The Rho family of small GTPases controls many biological processes, including cytoskeletal regulation, membrane trafficking, cell adhesion, cell polarization, transcriptional activity, apoptosis, and cell proliferation. Wrch1, which belongs to the Cdc42 subfamily, is one of the least characterized family member. Despite its homology to other Cdc42-like proteins, we found that Wrch1 has unique characteristics. Biochemical experiments showed that Wrch1 has no detectable GTPase activity in vitro and that its intrinsic nucleotide exchange rate is very high in comparison to Cdc42. Furthermore, NIH3T3 cells transiently transfected with Wrch1 showed an up-rounded, retracted phenotype. In addition, Wrch1 was shown to be more efficient than Cdc42 in triggering the formation of filopodia. Serum stimulation of cells expressing Wrch1 induces vigorous membrane blebbing, a phenomenon dependent on the activity of ROCK. In a search for proteins interacting with Wrch1, PAK1 and NCKbeta were identified as binding partners. Interestingly, the interaction to NCKbeta was shown to be mediated via PxxP motifs present in an N-terminal extension of Wrch1 to the second and third SH3 domains of NCKbeta. PMID- 15350536 TI - Phosphatidylcholine-specific phospholipase C in mitogen-stimulated fibroblasts. AB - To investigate expression, subcellular localization and mechanisms of translocation of phosphatidylcholine-specific phospholipase C (PC-PLC) during the cell proliferative response, biochemical, immunoblotting, and immunofluorescence analyses were performed on quiescent and mitogen-stimulated NIH-3T3 fibroblasts. Platelet-derived growth factor (PDGF), insulin and 12-O-tetradecanoylphorbol-13 acetate induced, in 10-60 min, PC-PLC translocation from a perinuclear cytoplasmic area to the plasma membrane. Following cell exposure to PDGF (60 min), the overall PC-PLC expression increased up to 2-3x, while the enzyme activity increased 5x in total cell lysates, 2x in the plasma membrane, and 4x in the nucleus; moreover, confocal laser scanning microscopy showed a progressive externalization of PC-PLC on the outer plasma membrane surface and its accumulation in the nuclear matrix. Pre-incubation of cells with the PC-PLC inhibitor tricyclodecan-9-yl potassium xanthate (D609), before PDGF-stimulation, not only reduced the enzyme activity in total cell lysates as well as in plasma membrane and nuclear fractions, but also blocked the mechanisms of PC-PLC subcellular redistribution. These effects were associated with a D609-induced long-lasting cell cycle block in Go. PMID- 15350537 TI - Bone marrow stromal proteoglycans regulate megakaryocytic differentiation of human progenitor cells. AB - Adherence of hematopoietic progenitor cells (HPCs) to stroma is an important regulatory step in megakaryocytic differentiation. However, the mechanisms through which megakaryocytic progenitors are inhibited by stroma are poorly understood. We examined the role of sulfated glycoconjugates, such as proteoglycans (PGs), on human bone marrow stroma (hBMS). To this end, PG structure was altered by desulfation or enzymatic cleavage. PGs participated in adhesion of human HPC, as desulfation resulted in about 50% decline in adhesion to hBMS. Heparan sulfate proteoglycans (HSPGs) were found to be responsible by showing about 25% decline in adhesion after pre-incubation of HPC with heparin and about 15% decline in adhesion after enzymatic removal of HSPGs from hBMS. Furthermore, PGs were involved in binding cytokines. Both desulfation and enzymatic removal of stromal HSPGs increased release of megakaryocytopoiesis inhibiting cytokines, that is, interleukin-8 (IL-8, 1.9-fold increase) and macrophage inflammatory protein-1alpha (MIP-1alpha, 1.4-fold increase). The megakaryocytic output of HPC grown in conditioned medium of desulfated stroma was decreased to 50% of the megakaryocytic output in CM of sulfated stroma. From these studies, it can be concluded that PGs in bone marrow, in particular HSPGs, are involved in binding HPC and megakaryocytopoiesis-inhibiting cytokines. Bone marrow stromal PGs thus reduce differentiation of HPC toward megakaryocytes. PMID- 15350538 TI - Heat stress-induced localization of small heat shock proteins in mouse myoblasts: intranuclear lamin A/C speckles as target for alphaB-crystallin and Hsp25. AB - We examined the effect of heat stress on localization of two sHsps, alphaB crystallin and Hsp25, and of Hsc70, a member of a different class of heat shock proteins (Hsps), in both undifferentiated and differentiated mouse C2C12 cells. Under normal conditions, alphaB-crystallin and Hsp25 are found in the cytoplasm; only alphaB-crystallin is also found in the nucleus, distributed in a speckled pattern. Hsc70 is found to be homogeneously distributed throughout the cell. On heat stress, all these proteins translocate almost entirely into the nucleus and upon recovery relocate to the cytoplasm. Dual staining experiments using C2C12 myoblasts show that alphaB-crystallin and Hsp25, but not Hsc70, colocalize with the intranuclear lamin A/C and the splicing factor SC-35, suggesting interactions of sHsps and intranuclear lamin A/C. Interestingly, none of these proteins are found in the myotube nuclei. Upon heat stress, only Hsc70 translocates into the myotube nuclei. This differential entry of alphaB-crystallin and Hsp25 into the nuclei of myoblasts and myotubes upon heat stress may have functional role in the development and/or in the maintenance of muscle cells. Our study therefore suggests that these sHsps may be a part of the intranuclear lamin A/C network or stabilizing this specific network. PMID- 15350539 TI - Actin overexpression parallels severity of pancreatic injury. AB - Among the three major cytofilament proteins, keratin (K8/K18/K19) expression increases nearly threefold upon pancreas or liver injury, while actin and tubulin expressions are considered relatively stable. K8/K18 serves essential hepatocyte cytoprotective functions yet appears dispensable in K8-null mouse pancreata, which led us to hypothesize that actin or tubulin expressions may increase after pancreatic injury. Balb/c and FVB/n mice manifested different susceptibility to injury in two pancreatitis models, with significant induction of actin protein (threefold) and RNA after moderate or severe but not mild injury. Alterations in tubulin expression were less prominent. Basally, K8-null and wild-type pancreata expressed similar actin and tubulin levels, while the injury-induced actin protein but not RNA was more pronounced in K8-null mice. K7/K18/K19/K20 were also induced in K8-null mice after injury. Ex vivo, caerulein-triggered pancreatitis caused protein degradation (actin approximately or = tubulin > keratins) and mRNA up-regulation that was blocked by actinomycin-D (act-D) (actin approximately or = tubulin approximately or = keratin) or by NF-kappaB inhibition (keratins > actin approximately or = tubulin). Hence, actin is not as static as previously held and is overexpressed after moderate to severe pancreatic injury while keratins are induced after minimal injury. Keratin and actin induction may serve protective roles in pancreatic injury. PMID- 15350540 TI - Simultaneous cell death and desquamation of the embryonic diffusion barrier during epidermal development. AB - The periderm is an epithelial layer covering the emerging epidermis in early embryogenesis of vertebrates. In the chicken embryo, an additional cellular layer, the subperiderm, occurs at later embryonic stages underneath the periderm. The questions arose what is the function of both epithelial layers and, as they are transitory structures, by which mechanism are they removed. By immunocytochemistry, the tight junction (TJ) proteins occludin and claudin-1 were localized in the periderm and in the subperiderm, and sites of close contact between adjacent cells were detected by electron microscopy. Using horseradish peroxidase (HRP) as tracer, these contacts were identified as tight junctions involved in the formation of the embryonic diffusion barrier. This barrier was lost by desquamation at the end of the embryonic period, when the cornified envelope of the emerging epidermis was formed. By TUNEL and DNA ladder assays, we detected simultaneous cell death in the periderm and the subperiderm shortly before hatching. The absence of caspases-3, -6, and -7 activity, key enzymes of apoptosis, and the lack of typical morphological criteria of apoptosis such as cell fragmentation or membrane blebbing point to a special form of programmed cell death (PCD) leading to the desquamation of the embryonic diffusion barrier. PMID- 15350541 TI - IL-1beta regulates expression of Cx32, occludin, and claudin-2 of rat hepatocytes via distinct signal transduction pathways. AB - The functions of gap and tight junctions are perturbed during the acute-phase response to liver injury. To elucidate the mechanism of pro-inflammatory cytokine IL-1beta responsible for regulation of hepatic gap and tight junctions, we analyzed expression and function of gap and tight junctions using a rat liver injury model and primary cultures of rat hepatocyte. In rat liver lobules at 24 h after thioacetamide (TAA) treatment, where some IL-1beta-positive non-parenchymal cells existed, disappearance of connexin32-positive spots at cell borders of the hepatocytes and increases of claudin-2 and occludin immunoreactivities in bile canalicular regions were observed. In primary cultures of rat hepatocytes, IL 1beta caused the disappearance of connexin32, which was reciprocal to the induction and localization of claudin-2 to cell membranes. The downregulated connexin32 expression was inhibited by treatment with a MAP-kinase inhibitor (PD98059), whereas the upregulated claudin-2 expression was blocked by p38 MAP and PI3-kinase inhibitors (SB203580 and LY294002). The changes of connexin32 and claudin-2 may be controlled at the transcriptional level via NF-kappaB, HNF 1alpha, and CDX2. Occludin was hyperphosphorylated by IL-1beta treatment and was inhibited by treatment with a PI3-kinase inhibitor. These results demonstrate that MAP-kinase, p38 MAP-kinase, and PI3-kinase are distinctly involved in the regulation of hepatic gap and tight junctions during the acute-phase response to IL-1beta. PMID- 15350542 TI - Suppression of MeCP2beta expression inhibits neurite extension in PC12 cells. AB - Regulation of gene expression is critical to the proper development of neuronal cells. The methyl-CpG binding protein 2 (MeCP2) operates as a transcriptional repressor by facilitating histone deacetylation and DNA methylation-dependent transcriptional silencing. This study examined the importance of MeCP2 in the regulation of neurite formation in PC12 cells. Expression of MeCP2 increased in a time-dependent manner after induction of neuronal differentiation. Expression was assessed at both the transcriptional and translation levels, and reached a maximum at 24 h post-induction. In addition, a marked inhibition of neurite extension and proper localization of a marker for synapse formation, synapsin I, were observed when MeCP2 expression was decreased by the addition of an antisense morpholino oligomer directed to the translational initiation site for MeCP2beta. The removal of the antisense oligomer allowed neurite extension to progress. However, the addition of antisense oligomer to previously differentiated PC12 cells did not affect established neurite processes. Taken collectively, our results indicate a role for MeCP2beta early in the events of neurite formation and that the relative levels of MeCP2alpha and MeCP2beta may be different in early differentiating neurons than is found in the adult brain. In addition, unique functions may exist for the two isoforms of MeCP2. Our results indicate that the inhibition of neurite elaboration caused by a reduction in MeCP2 may be reversible. PMID- 15350543 TI - CADASIL-associated Notch3 mutations have differential effects both on ligand binding and ligand-induced Notch3 receptor signaling through RBP-Jk. AB - Mutations in the NOTCH3 gene are the cause of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a hereditary angiopathy leading to strokes and dementia. Pathogenic mutations remove or insert cysteine residues within epidermal growth factor (EGF) repeats in the extracellular domain of the Notch3 receptor (N3ECD). Vascular smooth muscle cells (VSMC) are the predominant site of Notch3 expression in adults. In CADASIL patients, VSMC degenerate and N3ECD is deposited within the vasculature. However, the mechanisms underlying VSMC degeneration and N3ECD accumulation are still unknown. In this study, we investigated the consequences of three pathogenic Notch3 mutations on the biological activity of the receptor by analyzing ligand (Delta-/Jagged-)-induced signaling via RBP-Jk. Two mutations (R133C and C183R) that are located outside the putative ligand binding domain (LBD) of the receptor were found to result in normal Jagged1-induced signaling in A7r5 VSMC, whereas the third mutation (C455R located within the putative LBD) showed strongly reduced signaling activity. Ligand binding assays with soluble Delta1 and Jagged1 revealed that C455R interferes with ligand binding through disruption of the LBD which, as we show here, is located in EGF repeats 10/11 of Notch3. All mutant receptors including Notch3C455R were targeted to the cell surface but showed an elevated ratio between the unprocessed full-length 280-kDa receptor and S1-cleaved receptor fragments. Taken together, these data indicate that CADASIL-associated Notch3 mutations differ with respect to their consequences both on ligand binding and ligand-induced signaling through RBP-Jk, whereas they have similar effects on receptor maturation. Moreover, the data suggest that ligand-induced receptor shedding may not be required for N3ECD deposition in CADASIL. PMID- 15350544 TI - Matrix metalloproteinase inhibitor GM 6001 attenuates keratinocyte migration, contraction and myofibroblast formation in skin wounds. AB - In this study, we examined the impact of matrix metalloproteinases (MMP) on epithelialization, granulation tissue development, wound contraction, and alpha smooth muscle actin (ASMA) expression during cutaneous wound repair through systemic administration of the synthetic broad-spectrum MMP inhibitor GM 6001 (N [(2R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl]-L-tryptophan methylamide). Four full-thickness excisional wounds (50 mm2) on the back of 22 young female Sprague-Dawley rats, 12 treated with GM 6001 100 mg/kg and 10 with vehicle, were allowed to heal by secondary intention. GM 6001-treated wounds were minimally resurfaced with neoepithelium, despite unaltered keratinocyte proliferation in wound edges, whereas control wounds were completely covered with 3-7 cell layers of parakeratinized epithelium on post-wounding day 7. Hydroxyproline concentration, a marker of collagen, and cell proliferation in granulation tissue did not differ significantly between GM 6001-treated and control groups. Impaired wound contraction (P < 0.01) was associated with a dramatic reduction of ASMA positive myofibroblasts in granulation tissue of GM 6001 wounds. This was not due to GM6001 blocking transforming growth factor-beta1 (TGF-beta1)-induced myofibroblast differentiation since GM 6001 did not inhibit TGF-beta1-induced ASMA expression and force generation in cultured rat dermal fibroblasts. The profound impairment of skin repair by the nonselective MMP inhibitor GM 6001 suggests that keratinocyte resurfacing, wound contraction, and granulation tissue organization are highly MMP-dependent processes. PMID- 15350545 TI - Differential activation of vascular genes by hypoxia in primary endothelial cells. AB - Changes in the local environment, such as reduced oxygen tension (hypoxia), elicit transcriptional activation of a variety of genes in mammalian cells. Here we have analyzed the effect of hypoxia in different vascular endothelial cells (ECs) with emphasis on hypoxia-regulated transcription factors and genes of importance for blood vessel dynamics. While hypoxia induced the transcription factor hypoxia-inducible factor-1alpha (HIF-1alpha) in all endothelial cells tested, the closely related HIF-2alpha protein was markedly induced in microvascular/capillary endothelial cells, but only weakly or not at all in artery and vein endothelial cells. Furthermore, microvascular/capillary endothelial cells responded to hypoxia with increased number of transcripts encoding vascular endothelial growth factor-A (VEGF-A), VEGF receptor-2, the angiopoietin receptor Tie2, platelet-derived growth factor-B (PDGF-B), and inducible nitric oxide synthase (iNOS). In vein endothelial cells, hypoxia instead increased transcripts encoding lymphatic vascular components VEGF-C, -D, and VEGF receptor-3. Finally, reduced VEGF receptor levels and phosphorylation indicated establishment of a functional autocrine VEGF-A loop in hypoxic endothelial cells. Our results show that endothelial cells, derived from different vascular beds, mount different transcriptional responses to changes in oxygen tension. PMID- 15350546 TI - Formation of aggresome-like structures in herpes simplex virus type 2-infected cells and a potential role in virus assembly. AB - Herpes simplex virus (HSV) is a large, enveloped DNA virus that replicates in the nucleus and is assembled in the cytoplasm to the mature infectious virion. In this study, we present evidence that, in HSV-2-infected cells, some tegument proteins (UL46 and VP16) and newly synthesized nucleocapsids accumulate in a juxtanuclear domain sharing characteristics with aggresomes, cellular structures formed in response to misfolded proteins [J. Cell Biol. 146 (1999) 1239, J. Cell Biol. 143 (1998) 2010]. The juxtanuclear domains (aggresome-like structures) induced by HSV-2 infection localize to the microtubule organizing center (MTOC) where the clustering mitochondria, Golgi-derived vesicles, and cellular chaperones including heat shock protein (Hsp)40 and Hsp70 were recruited. Formation of aggresome-like structures was blocked by the presence of microtubule disassembling drug nocodazole, indicating that microtubule-dependent transport may be involved in the accumulation of viral and cellular proteins at these sites in HSV-2-infected cells. These features are similar to those governing the formation of aggresomes. In contrast to aggresomes, however, the vimentin cage surrounding the MTOC was not observed with the aggresome-like structures in HSV-2 infected cells, and the maintenance of these structures required an intact microtubular network. Disruption of the aggresome-like structures by nocodazole treatment led to a low but consistent effect (10-fold decrease) on the production of intracellular infectious particles. These results suggest that aggresome-like structures do not play a critical but augmentary role in HSV-2 replication. PMID- 15350547 TI - Early enterocytic differentiation of HT-29 cells: biochemical changes and strength increases of adherens junctions. AB - We have characterized the modulation of cell-cell adhesion and the structure of adherens junctions in the human colon adenocarcinoma HT-29 cell line that differentiates into enterocytes after glucose substitution for galactose in the medium. We demonstrate that differentiated cells (HT-29 Gal) rapidly established E-cadherin-mediated interactions in aggregation assays. This effect is not due to an increase in E-cadherin expression during this early stage of cell differentiation, but rather results from the maturation of preexisting adherens junctions. These junctions are characterized by the redistribution of E-cadherin to the basolateral membrane and its co-localization with the actin cytoskeleton. Subcellular fractionation studies indicate that actin-associated E-cadherins bind beta-catenin and p120ctn. Furthermore, the p120ctn/E-cadherin association is upregulated. These data reveal a cooperative interaction between p120ctn and E cadherin that corresponds to mature functional adherens junctions able to initiate tight cell-cell adhesion required for epithelium architecture and further affirm the gatekeeper role of p120ctn. PMID- 15350548 TI - Vasoactive intestinal peptide-induced neurite remodeling in human neuroblastoma SH-SY5Y cells implicates the Cdc42 GTPase and is independent of Ras-ERK pathway. AB - Vasoactive intestinal peptide (VIP) is known to regulate proliferation or differentiation in normal and tumoral cells. SH-SY5Y is a differentiated cell subclone derived from the SK-N-SH human neuroblastoma cell line and possess all the components for an autocrine action of VIP. In the present study, we investigated the morphological changes and intracellular signaling pathways occurring upon VIP treatment of SH-SY5Y cells. VIP induced an early remodeling of cell projections: a branched neurite network spread out and prominent varicosities developed along neurites. Although activated by VIP, the Ras/ERK pathway was not required for the remodeling process. In contrast, pull-down experiments revealed a strong Cdc42 activation by VIP while expression of a dominant-negative Cdc42 prevented the VIP-induced neurite changes, suggesting an important role for this small GTPase in the process. These data provide the first evidence for a regulation of the activity of Rho family GTPases by VIP and bring new insights in the signaling pathways implicated in neurite remodeling process induced by VIP in neuroblastoma cells. PMID- 15350549 TI - E1AF/PEA3 reduces the invasiveness of SiHa cervical cancer cells by activating serine proteinase inhibitor squamous cell carcinoma antigen. AB - E1AF/PEA3, a member of the Ets family of transcription factors, is associated with the malignant characteristics of cancer cells. The initial aim of our study was to test whether the invasiveness of SiHa cervical cancer cells could be diminished by transfection with antisense E1AF. Using an in vitro invasion assay in which cells penetrate a layer of Matrigel, we found that this was not the case; indeed, the invasiveness of the transfectants was enhanced. To better understand the mechanism of this enhancement, we used the cDNA microarray technique to search for genes whose expression was altered in the antisense E1AF transfected SiHa cells. Among several genes affected, we found that expression of squamous cell carcinoma antigen (SCCA), a member of the ovalbumin serine proteinase inhibitor family, was significantly reduced. Forced expression of E1AF enabled activation of SCCA expression, and Luciferase reporter assays revealed that E1AF activates the SCCA promoter. Introduction of antisense SCCA into SiHa cells inhibited production of SCCA protein and markedly increased the invasiveness of the cells. Taken together, these results suggest that E1AF suppresses the invasiveness of SiHa cervical cancer cells through transcriptional activation of the SCCA serine proteinase inhibitor gene. PMID- 15350550 TI - Bcl-2 prevents loss of mitochondria in CCCP-induced apoptosis. AB - Bcl-2 family proteins regulate apoptosis at the level of mitochondria. To examine the mechanism of Bcl-2 function, we investigated the effects of the protonophore carbonyl cyanide m-chlorophenyl hydrazone (CCCP) on two hematopoietic cell lines and Bcl-2 overexpressing transfectants. CCCP directly interferes with mitochondrial function and induces apoptosis. We show that Bcl-2 inhibits apoptosis and that the antiapoptotic effect of Bcl-2 takes place upstream of caspase activation and nuclear changes associated with apoptosis, since these were markedly inhibited in cells overexpressing Bcl-2. Bcl-2 does not prevent the decrease in mitochondrial membrane potential nor the alterations in cellular ATP content induced by CCCP in FL5.12 and Jurkat cells. A higher number of mitochondria was observed in untreated Bcl-2 transfected cells compared to parental cells, as shown by electron microscopy. Exposure to CCCP induced a dramatic decrease in the number of mitochondria and severely disrupted mitochondrial ultrastructure, with apparent swelling and loss of cristae in parental cells. Bcl-2 clearly diminished the disruption of mitochondrial structure and preserved a higher number of mitochondria. These data suggest that CCCP induces apoptosis by structural disruption of mitochondria and that Bcl-2 prevents apoptosis and mitochondrial degeneration by preserving mitochondrial integrity. PMID- 15350551 TI - Hypercholesterolemia impairs basal nitric oxide synthase turnover rate: a study investigating the conversion of L-[guanidino-(15)N(2)]-arginine to (15)N-labeled nitrate by gas chromatography--mass spectrometry. AB - Endothelial function is impaired in hypercholesterolemia and atherosclerosis, which is probably due to reduced biological activity of endothelium-derived nitric oxide (NO). NO is synthesized in functionally intact endothelium by oxidation of the terminal guanidino nitrogen atom(s) of the amino acid precursor, L-arginine. We applied stable isotope dilution techniques and gas chromatographic mass spectrometric approaches to investigate metabolism of L-[guanidino-(15)N(2)] arginine to (15)N-labeled nitrate in hypercholesterolemic rabbits and controls. After 4 weeks on control or 1% cholesterol-enriched diet, rabbits received 267 +/ 6 micromol of L-[guanidino-(15)N(2)]-arginine/kg of body weight via gastric cannulation. (15)N-isotope content of L-arginine in plasma and in platelet lysates increased 2h later in both groups, and almost returned to baseline until 24h. (15)N-isotope content of plasma nitrite and nitrate also increased in both groups at 2h, and had almost returned to natural content 24h later. (15)N-isotope content of urinary nitrate was significantly increased in control animals in urines collected from 0 to 12, 12 to 24, and had returned to baseline in the urine sample collected from 24 to 48 h. In the cholesterol group only a slight, insignificant elevation of (15)N-isotope content was observed for urinary nitrate. The extent of conversion of L-[guanidino-(15)N(2)]-arginine to (15)N labeled nitrate was strongly and inversely correlated to plasma concentration of the endogenous NO synthase inhibitor, asymmetric dimethylarginine (ADMA), which was elevated in cholesterol-fed rabbits (R=0.77; p < 0.05). Our data show that baseline NO synthase turnover rate is reduced in rabbits during early hypercholesterolemia. Our study gives evidence that the mechanism of the impaired conversion of L-[guanidino-(15)N(2)]-arginine to (15)N-labeled nitrate most likely involves inhibition of NO synthase by ADMA, which is present in elevated concentrations in hypercholesterolemia. PMID- 15350552 TI - Cycloprodigiosin up-regulates inducible nitric oxide synthase gene expression in hepatocytes stimulated by interleukin-1beta. AB - The proinflammatory cytokine interleukin-1beta stimulates inducible nitric oxide synthase (iNOS) gene expression in hepatocytes via activation of a transcription factor, nuclear factor-kappaB (NF-kappaB). Nitric oxide has diverse cytoprotective and toxic effects. Cycloprodigiosin is an anticancer drug that induces apoptosis through NF-kappaB-dependent mechanisms. This study investigated whether cycloprodigiosin influenced NF-kappaB activation and induction of iNOS by interleukin-1beta. Cycloprodigiosin alone had no effect on NO production by primary cultured rat hepatocytes. Simultaneous addition of cycloprodigiosin and interleukin-1beta markedly stimulated the induction of iNOS mRNA and protein compared with addition of interleukin-1beta alone, resulting in overproduction of NO. Cycloprodigiosin had no effect on degradation of the inhibitory subunit of NF kappaB (IkappaBalpha), but lessened the recovery of IkappaBalpha levels. The electrophoretic mobility shift assay revealed that cycloprodigiosin caused an increase of NF-kappaB activation. Consistent with this observation, cycloprodigiosin promoted the translocation of p65 (a subunit of NF-kappaB) to the nucleus. Furthermore, this drug enhanced expression of the type 1 interleukin 1 receptor, and this action showed similar concentration-dependence to its induction of iNOS. These results indicate that cycloprodigiosin up-regulates the induction of iNOS by increasing NF-kappaB activation, at least partly through enhancement of type 1 interleukin-1 receptor expression. By regulating the expression of NF-kappaB-dependent genes, such as iNOS, cycloprodigiosin administration may increase NO production during hepatic injury. PMID- 15350553 TI - The effects of N(omega)-propyl-L-arginine on reperfusion injury of skeletal muscle. AB - N(omega)-Propyl-L-arginine (NPA) is reported to be a highly selective inhibitor of neuronal nitric oxide synthase (nNOS). This in vivo study observed its role in ischemia/reperfusion (I/R) injury in rat skeletal muscle. Our results showed that NPA infusion significantly increased vessel diameters and blood flow in reperfused cremaster muscle, and slightly increased contractile function in reperfused extensor digitorum longus (EDL) muscle. In addition, NPA treatment slightly increased I/R-mediated downregulation of nNOS and eNOS mRNA and protein levels. Although NPA showed a beneficial role in I/R injury, our in vivo data do not support NPA as a selective nNOS inhibitor. Also, our data do not provide any insight into the mechanism of NPA. Thus, the in vivo mechanism of action of NPA needs to be further identified, and the role of nNOS in skeletal muscle I/R still remains to be determined. PMID- 15350554 TI - A combination of statins and beta-blockers is independently associated with a reduction in the incidence of perioperative mortality and nonfatal myocardial infarction in patients undergoing abdominal aortic aneurysm surgery. AB - OBJECTIVE: To investigate the combined beneficial effect of statin and beta blocker use on perioperative mortality and myocardial infarction (MI) in patients undergoing abdominal aortic aneurysm surgery (AAA). BACKGROUND: Patients undergoing elective AAA-surgery identified by clinical risk factors and dobutamine stress echocardiography (DSE) as being at high-risk often have considerable cardiac complication rate despite the use of beta-blockers. METHODS: We studied 570 patients (mean age 69+/-9 years, 486 males) who underwent AAA surgery between 1991 and 2001 at the Erasmus MC. Patients were evaluated for clinical risk factors (age>70 years, histories of MI, angina, diabetes mellitus, stroke, renal failure, heart failure and pulmonary disease), DSE, statin and beta blocker use. The main outcome was a composite of perioperative mortality and MI within 30 days of surgery. RESULTS: Perioperative mortality or MI occurred in 51 (8.9%) patients. The incidence of the composite endpoint was significantly lower in statin users compared to nonusers (3.7% vs. 11.0%; crude odds ratio (OR): 0.31, 95% confidence interval (CI): 0.13-0.74; p=0.01). After correcting for other covariates, the association between statin use and reduced incidence of the composite endpoint remained unchanged (OR: 0.24, 95% CI: 0.10-0.70; p=0.01). Beta blocker use was also associated with a significant reduction in the composite endpoint (OR: 0.24, 95% CI: 0.11-0.54). Patients using a combination of statins and beta-blockers appeared to be at lower risk for the composite endpoint across multiple cardiac risk strata; particularly patients with 3 or more risk factors experienced significantly lower perioperative events. CONCLUSIONS: A combination of statin and beta-blocker use in patients with AAA-surgery is associated with a reduced incidence of perioperative mortality and nonfatal MI particularly in patients at the highest risk. PMID- 15350555 TI - Intra-mucosal acidosis as a predictor of cardiac outcome following abdominal aortic aneurysm surgery. AB - OBJECTIVE: To assess if sigmoid ischaemia is a prognostic indicator of early and late post-operative cardiac morbidity and mortality. MATERIALS AND METHODS: Patients undergoing elective abdominal aortic aneurysm repair (AAAR) were included in the study. Demographic details and risk factors for heart disease were recorded. Sigmoid pHi was measured at the time of surgery using a silicone tonometer and perioperative morbidity and mortality were recorded in all patients. Seven years following surgery the patients and their general practitioners were contacted to determine the patient's health. RESULTS: Thirty eight patients were included in the study. Within the follow-up period, 22 (58%) had died. Eight patients died of cardiac failure or myocardial infarction. The pHi in patients with cardiac related deaths [6.99 (6.84-7.10)] was significantly lower than those with non-cardiac related deaths [7.11 (7.04-7.21), p<0.05]. Similarly, patients who suffered acute cardiac events (within 30 days following AAA repair) had lower pHi [7.01 (6.88-7.12)] compared to those who did not [7.09 (6.90-7.19), p<0.05]. CONCLUSION: The results show that sigmoid ischaemia is more frequent amongst patients that develop cardiac events after AAAR and is associated with a worse long term outcome. This suggests that global hypoperfusion as a result of an under performing heart may be partly responsible for the sigmoid ischaemia in patients following AAAR. Therefore, low sigmoid pHi may predict an increased risk of cardiac complications in these patients. PMID- 15350556 TI - Frequency and outcome of re-interventions after endovascular repair for abdominal aortic aneurysm: a prospective cohort study. AB - PURPOSE: To describe frequency, type, and outcome of re-intervention after endovascular aortic aneurysm repair (EVAR). METHODS: Between September 1996 and December 2003, 308 patients were treated, with data collected prospectively. No patient was lost to follow up, but two were excluded (one primary conversion, and one post-operative death). Vanguard, Talent, Excluder, Zenith, and Quantum devices were used. Follow up required a CT scan before discharge. Initially, a CT scan was done at each follow up. Subsequently, we used duplex ultrasound and abdominal X-ray, with CT scan used selectively. RESULTS: Mean follow-up was 36+/ 22 months. Re-interventions were required in 47 (15%) patients, 31 (66%) elective and 16 (34%) emergency cases. In 32 patients, the primary re-intervention was successful; in 15 patients an additional 13 secondary and four tertiary re interventions were required. A total of 72 adjunctive manoeuvres were performed: 49 endovascular (68%) and 23 open (32%). The success of endovascular re interventions was 80%. The success of open re-interventions was 96%. Open conversions were required in nine patients (3%). There was no mortality. CONCLUSION: EVAR was associated with a low burden of re-interventions, with only 15% patients requiring re-intervention. Our long-term follow up, without regular CT, was simple and effective. PMID- 15350557 TI - Endovascular versus open surgical repair of abdominal aortic aneurysm: a comparison of early and intermediate results in patients suitable for both techniques. AB - PURPOSE: To assess early and intermediate results of endovascular repair of abdominal aortic aneurysms (EVAR), and to compare them with open surgery (OS) in concurrent patients suitable for both types of treatment. METHODS: During 3 years, 180 patients with AAA underwent repair. We excluded patients with ruptured aneurysms (33), juxtarenal aneurysms (11), iliac aneurysms (8), with peripheral embolization (2) and those treated with a cryopreserved homograft (2). From the remaining patients (n=124), we selected those suitable for both techniques (n=83), of which 53 were treated by EVAR and 30 by OS. Analysis was performed using Kaplan-Meier curves and Log Rank tests. RESULTS: Hospital mortality was not significantly higher in the OS group (6.6% OS vs. 3.7% EVAR), p=0.55. The EVAR group had significantly shorter operative time, length of hospital stay and less blood loss. The median follow up time was 2.18 years for OR and 1.58 years for EVAR. There were no conversions from EVAR to OS and no differences in late survival (p=0.255, Cox regression analysis) with a cumulative survival rate at 3 years of 89% for EVAR and 73% for OS. By 3 years 24% (95% CI, 11-47%) of EVAR patients had presented endoleaks with an endovascular re-intervention rate of 27% (95% CI, 13-50%). One patient in the OS group needed a late open intervention. CONCLUSIONS: EVAR compares favourably with OS in terms of reduction of operative time, hospital length of stay and blood loss. This study did not show a difference in early or late mortality. EVAR durability remains the most critical issue to be addressed. PMID- 15350558 TI - Intrasac pressure waveforms after endovascular aneurysm repair (EVAR) are a reliable marker of type I endoleaks, but not type II or combined types: an experimental study. AB - PURPOSE: To ascertain the nature of the pressure wave transmitted through aneurysm thrombus and the changes produced after endovascular repair and the development of type I and II endoleaks. METHODS: A 25 mm Talent endovascular graft was deployed in a latex model of an abdominal aortic aneurysm, which was incorporated in a pulsatile flow unit. The graft was surrounded by thrombus analogue to simulate conditions in vivo. Pressure waveforms in the sac were captured over 5s at 1000 Hz in these settings: (i) no endoleaks (baseline), after introduction of (ii) type I (iii) type II and (iv) combined type I and II endoleaks. The arterial blood pressure settings used were 140/100 and 130/90 mmHg, denoted the high and low settings, respectively. ANOVA in Minitab 13 was applied for statistical analysis. RESULTS: Pulsatile waveforms were transmitted through the thrombus. Intrasac pressure after stent-grafting reduced to 110/107, 99/96 mmHg (p<0.001) (high, low settings, respectively). Introduction of a type I endoleak caused this to rise to 120/112, 115/107 mmHg (p<0.001, vs. baseline); after producing a type II endoleak these were 101/98, 91/88 mmHg (p<0.001, vs. baseline). A combined type I and II endoleak produced intrasac pressures identical to that of a type I endoleak. CONCLUSIONS: Intrasac pressure waveforms following EVAR are easily defined following a type I endoleak. Waveforms obtained following type II endoleak simulation resemble the baseline waveform in an attenuated form. Intrasac pressures are, therefore, a reliable marker for type I, but not a type II endoleak. In the case of a combined endoleak, the type I endoleak waveform effectively masks that of the type II. Intrasac thrombus faithfully transmits intrasac pressures. PMID- 15350559 TI - Erdosteine ameliorates neurological outcome and oxidative stress due to ischemia/reperfusion injury in rabbit spinal cord. AB - OBJECTIVE: Oxygen-derived free radicals have been suggested as important in degeneration after spinal cord ischemia. The aim of this study was to investigate whether erdosteine has a protective effect against spinal cord ischemia during aortic cross clamping. MATERIALS AND METHODS: New Zealand White rabbits (n=21) were divided into three groups. In the ischemia/reperfusion group (I/R) (n=8), the infrarenal aorta of rabbits was cross clamped for 21 min and then reperfused. In erdosteine group, the administration of erdosteine solution (50 mg/kg) was started two days before aortic cross-clamping and rabbits (n=8) were subjected to ischemia and reperfusion. Animals in control group (n=5) underwent a surgical procedure similar to the other groups but the aorta was not clamped. The animals were sacrificed at 72 h and histopathological, and biochemical analyses were carried out on the lumbar spinal cords. RESULTS: Erdosteine treatment was associated with improved neurological function in the postoperative period. Histopathological examination of spinal cord tissues in erdosteine group revealed changes consistent with mild ischemic injury, but rabbits in I/R group with paraplegia had total destruction of the motor neurons. Biochemical analyses of spinal cord tissues, in the I/R group, revealed a significant increase in the superoxide dismutase, xanthine oxidase, adenosine deaminase and myeloperoxidase activities, and a significant depletion in glutathione peroxidase activity when compared to that of control rabbits. Erdosteine treatment prevented the increase of all these enzymes except adenosine deaminase. Ischemia/reperfusion produced a significant increase in the tissue malondialdehyde levels. Ischemia/reperfusion induced increments in malondialdehyde content of the spinal cord were significantly prevented by erdosteine treatment. CONCLUSIONS: The present study demonstrated that erdosteine treatment before aortic cross clamping ameliorates neurological outcome, neuronal injury and oxidative stress in the rabbit spinal cord. PMID- 15350560 TI - Prevalence of varicose veins and venous anatomy in patients without truncal saphenous reflux. AB - OBJECTIVES: To determine the prevalence and distribution of primary venous reflux in the lower limbs in patients without truncal saphenous reflux. DESIGN: Prospective cohort study. PATIENTS AND METHODS: One thousand and seven hundred and twelve patients with suspected venous disease were examined by duplex ultrasonography. Seven hundred and thirty-five patients had primary varicose veins with competent saphenous trunks. Limbs with truncal saphenous reflux, deep vein reflux or obstruction, previous injection sclerotherapy or vein surgery, arterial disease and inflammation of non-venous origin were excluded from further consideration. The CEAP classification system was used for clinical staging. Systematic duplex ultrasound examination was undertaken to assess the distribution of incompetent saphenous tributaries. RESULTS: The prevalence of primary reflux with competent saphenous trunks was 43%. Reflux of GSV calf tributaries was the most common. The majority of the limbs (96%) belonged to chronic venous disease classes C1 and C2 of the CEAP classification. CONCLUSIONS: Superficial venous reflux causing varicose veins in the presence competent saphenous trunks is very prevalent in this series in contrast to other studies, presumably reflecting differing patient populations. Our data clearly show that varicose veins may occur in any vein and do not depend on truncal saphenous incompetence. Careful duplex ultrasound evaluation allows the pattern of venous reflux to be established in this group of patient ensuring appropriate management of varices. PMID- 15350561 TI - Preservation of venous valve function after catheter-directed and systemic thrombolysis for deep venous thrombosis. AB - OBJECTIVES: The aim of the study was to assess venous reflux and the obstruction pattern after catheter-directed and systemic thrombolysis of deep iliofemoral venous thrombosis. PATIENTS: Thirty-two patients treated either with systemic (16) or catheter-directed local thrombolysis (16) for massive iliofemoral thrombosis were identified from the hospital registry. METHODS: Clinical evaluation at follow up was based on the CEAP classification and disability score. Reflux was assessed by colour duplex ultrasonography and standardised reflux testing. A vascular surgeon blinded to treatment established the clinical status of the lower limb following the previous DVT. RESULTS: Valvular competence was preserved in 44% of patients treated with catheter-directed thrombolysis compared with 13% of those treated with systemic thrombolysis (p=0.049, Chi squared). Reflux in any deep vein was present in 44% of patients treated by catheter-directed lysis compared with 81% of patients receiving systemic thrombolysis (p=0.03, Chi squared). Reflux in any superficial vein was observed in 25% vs. 63% of the patients, respectively (p=0.03, Chi squared). There were significantly more patients with venous insufficiency of classes C0-1 in the group treated with catheter-directed thrombolysis. CONCLUSION: In this clinical series venous valvular function was better preserved after iliofemoral DVT when treated with catheter-directed thrombolysis. PMID- 15350562 TI - Vacuum-assisted conservative treatment for the management and salvage of exposed prosthetic hemodialysis access. AB - Recurrent puncture of dialysis grafts can cause erosion and ulcer formation in the skin over the prosthetic material. Contamination of the wound can lead to infection of the graft, and the necessity to remove it. We describe four cases where agressive treatment with debridement, intravenous antibiotics and negative pressure therapy allowed prosthesis salvage without discontinuation of hemodialysis. PMID- 15350563 TI - The management of short saphenous varicose veins: a survey of the members of the vascular surgical society of Great Britain and Ireland. AB - OBJECTIVE: The outcome of short saphenous vein surgery is often unsatisfactory and the high litigation rate reflects this. The aim of this study was to explore the current management of short saphenous varicose veins in Great Britain and Ireland. METHODS: This was a postal questionnaire survey of the surgical members of the Vascular Surgical Society of Great Britain and Ireland. Of 532 questionnaires 379 were returned (71.2%). RESULTS: There was diversity of opinion about the management of short saphenous veins. Eighty nine per cent of surgeons requested duplex imaging for all patients and over 50% arranged additional duplex marking of the saphenopopliteal junction preoperatively. Only 10.4% formally exposed and identified the popliteal vein during saphenopopliteal ligation, the majority (75.7%) dissected down the short saphenous vein to visualise the junction. The short saphenous vein was stripped routinely by 14.5% of surgeons, the majority preferring to excise a proximal segment of up to 10 cm (55.1%). Compared with long saphenous vein surgery, surgeons were generally more likely to warn patients of nerve damage but equally likely to warn of deep vein thrombosis. A small number of surgeons failed to warn patients of these complications. CONCLUSION: The variation in management of short saphenous veins may be explained by the lack of definitive clinical trials in this area. PMID- 15350564 TI - Healing of implanted expanded polytetrafluoroethylene vascular access grafts with different internodal distances: a histologic study in dogs. AB - OBJECTIVE: We assessed characteristics of healing, over time, of two types of expanded polytetrafluoroethylene grafts. STUDY DESIGN: An experimental histological study in dogs. METHODS: The graft types studied had the same internal diameter (5 mm) but different internodal distances. In one, the internodal distance was 60 microm in the external surface and 20 microm in the luminal surface. In the other, the internodal distance was 30 microm throughout the material. Sixteen grafts of each type were implanted between the femoral artery and vein in 16 dogs; explanted 1, 2, 4 or 12 weeks later; and examined histologically. RESULTS: In both graft types, infiltrating-cell density and maximum cell-penetration depth increased significantly between 1 and 2 weeks after implantation, but no significant increases occurred after 2 weeks. The number of inflammatory cells peaked 1 week after implantation and decreased significantly by 2 weeks. Subsequently, there were no significant changes in inflammatory cell numbers, suggesting that the inflammatory phase was over by 2 weeks after implantation and the grafts had become attached to surrounding tissue. There were no significant differences between the two graft types in cell density, cell-penetration depth, or number of inflammatory cells at any assessment time. CONCLUSION: Our results provide histologic support for guidelines recommending that synthetic vascular grafts for hemodialysis access should not be cannulated until 2 weeks after implantation. Since increasing the internodal distance to 60 microm in the external surface had no effect on graft healing, methods other than manipulation of internodal distance should be used in developing a graft suitable for early cannulation. PMID- 15350565 TI - Homocysteine levels, haemostatic risk factors and patency rates after endovascular treatment of the above-knee femoro-popliteal artery. AB - OBJECTIVES: To investigate the relationship between plasma homocysteine and other haemostatic variables and restenoses or reocclusions after endovascular treatment of symptomatic atherosclerosis of the above-knee femoro-popliteal artery. DESIGN: Prospective observational study. SETTING: University hospital. PATIENTS AND METHODS: The study included 103 patients (116 limbs), treated with subintimal angioplasty in 58 cases (50%) and with intraluminal PTA in 58 (50%): 39 (34%) patients were treated for critical limb ischaemia. Blood samples for analyses of fasting plasma values of homocysteine, fibrinogen, D-dimer, activated protein C resistance were drawn upon admission. Median follow-up for all procedures was 11 months (range 0-42 months). Outcome events (arterial patency) were defined as > or =50% restenosis or reocclusion in the treated arterial segment. Patency rates were estimated with the product limit method and Kaplan-Meier curves. Variables found to be related significantly to patency were included in multivariate analysis performed with the Cox proportional hazard model. RESULTS: The 1-year cumulative primary patency rate for all procedures was 48%. One-year limb salvage rate in cases of critical ischaemia was 74%. Multivariate analysis demonstrated significant independent associations between patency rates and plasma D-dimer, diabetes mellitus, the nature of the lesion treated (stenosis vs. occlusion) and antithrombotic therapy with aspirin after the procedure. Plasma levels of homocysteine, fibrinogen or activated protein C resistance were not associated with patency rates. Homocysteine levels were higher in patients with critical limb ischaemia than those with intermittent claudication. CONCLUSIONS: Early restenosis or reocclusion after endovascular intervention of lesions in the above knee femoro-popliteal artery was more frequent following treatment of occlusion (versus stenosis), for patients with diabetes, patients with elevated D-dimer and those without antithrombotic therapy after the procedure. Plasma homocysteine did not appear to influence the outcome of endovascular intervention. PMID- 15350566 TI - Seasonal variation of Buerger's disease in Northern part of Thailand. AB - OBJECTIVES: Buerger's disease is a common peripheral arterial occlusive disease in Asia, Middle East, and eastern European countries. This study was undertaken to investigate the seasonal variation in admission pattern of with patients Buerger's disease at our institution which is a referral hospital in the Northern Thailand. MATERIAL AND METHODS: Patients with Buerger's disease admitted to Chiang Mai University Hospital between January 1987 and December 2002 were studied retrospectively. Data are reported as mean+/-SD. Statistical significance was analyzed by Chi-square test. RESULTS: Eighty-four patients (82 men and two women) with Buerger's disease were evaluated on 121 admissions. Forty-five were newly diagnosed cases, who were admitted for initial treatment, and 39 were known cases who experienced worsening of the disease. Sixty-three admissions (52%) took place during winter (November to February), 44 admissions (34.6%) during the rainy season (June-October) and only 14 admissions (11.6%) occurred during the summer (March-May). There was a significant difference in the monthly admission rates during the three seasons (p<0.05). CONCLUSION: Admission for Buerger's disease showed a significant seasonal variation, with a peak in the winter followed by the rainy and summer season, respectively. Further research is needed to confirm our findings and evaluate the underlying mechanisms. PMID- 15350567 TI - Incidence of cranial nerve injuries after carotid eversion endarterectomy with a transverse skin incision under regional anaesthesia. AB - OBJECTIVES: The objective of this prospective study was to evaluate the incidence and distribution of cranial nerve injuries after carotid eversion endarterectomy (EEA) performed under regional anaesthesia using a transverse skin incision. PATIENTS AND METHODS: The study included 165 patients and 180 carotid arteries. All patients had a standard pre-operative assessment performed by a neurologist and ENT specialist. All carotid endarterectomies were performed by the eversion technique under regional anaesthesia. RESULTS: Ten cranial nerve injuries were observed. Seven patients had injuries of the marginal mandibular branch of the facial nerve, two patients had lesions of the hypoglossal nerve, and one patient had an injury of the recurrent laryngeal nerve. Eleven patients developed hoarseness without cranial nerve injury. Injuries of the marginal mandibular branch recovered after 3-8 months (mean 5.2 months). Both hypoglossal nerve injuries recovered after 4 months. The patient with the recurrent laryngeal palsy had no improvement after 19 months. Patients with hoarseness secondary to laryngeal haematoma recovered within 1 month. CONCLUSION: The incidence of cranial nerves injury after carotid EEA under regional anaesthesia is comparable to that reported for conventional carotid surgery. Postoperative hoarseness is most frequently due to laryngeal haematoma. PMID- 15350568 TI - The role of clinical examination in excluding vascular injury in haemodynamically stable patients with gunshot wounds to the neck. A prospective study of 59 patients. AB - OBJECTIVE: To prospectively evaluate the safety and accuracy of physical examination in determining the management of stable patients with gunshot wounds to the neck. DESIGN: Prospective study of 59 patients with gunshot wounds to the neck. PATIENTS AND METHODS: Fifty-nine stable patients with gunshot wounds to the neck managed between December 2001 and August 2003. All patients had a physical examination and routine angiography according to a written protocol approved by the research ethics committee. The sensitivity, specificity, and predictive values of physical examination were assessed and compared with the angiographic findings. RESULTS: Thirteen patients with positive findings on physical examination (history of bleeding, haematoma, minimal bleeding, thrill, bruit and pulse deficit) and 10 patients without clinical signs of vascular injury had vascular injury. A sensitivity of 57%, specificity 53%, positive predictive value 43% and negative predictive value of 67% were calculated for physical examination alone in detecting vascular injury. CONCLUSION: Findings on physical examination are not good predictors of vascular injury in stable patients with gunshot wounds to the neck. Our findings question the validity of physical examination alone, as a safe and accurate assessment of patients with gunshot wounds to the neck. Arteriography or ultrasonography is needed to identify vascular injuries. PMID- 15350569 TI - Compliance matching stent placement in the carotid artery of the swine promotes optimal blood flow and attenuates restenosis. AB - OBJECTIVES: We assessed the value of a gradient-compliant stent in an animal model. METHODS: Bilateral carotid arteries were stented with nitinol stents having variable-oversizing, variable-stiffness, and with (CMS, 10 animals) and without (SMART, four animals) compliance-matching endings. Angiography, hemodynamic, scanning-electron-microscopic and histological analyses were performed at 3-month. The protocol was completed in 14 among 19 swines. RESULTS: Transient (1-month) exaggerated recoil, attributable to stress-induced phasic inhibition of vasorelaxation, developed at CMS endings. At mid-term, all stents were endothelialized; CMS-stents, but not SMART-stents, were incorporated into walls (one-strut-thickness). Restenosis developed outside SMART-stents (cell migration+wall-compensatory enlargement) whereas CMS-stents elicited no or focalized cell-accumulations at endings that bulged vascular walls radially outward. SMART-stents were blood-flow neutral, whereas CMS-stents favored (higher stiffness, higher-oversizing) or opposed (lower-stiffness, less-oversizing) carotid blood flow. CONCLUSIONS: Direct carotid stenting with stents having compliance-matched endings and specific requirements of stiffness and oversizing can optimize blood flow to the brain and restrict local restenosis. PMID- 15350570 TI - 1603-2003: Four centuries of valves. PMID- 15350571 TI - Simultaneous endovascular exclusion of thoracic aortic aneurysm with open abdominal aortic aneurysm repair. AB - BACKGROUND: The treatment of aneurysms at multiple sites within the aorta is problematic. METHODS: Between March 2002 and June 2003 in the Department of General, Vascular and Transplant Surgery, Medical University of Warsaw six patients with coexisting abdominal and descending thoracic aortic aneurysms underwent simultaneous open abdominal aortic aneurysm (AAA) repair and endoluminal thoracic aortic aneurysm (TAA) repair. The indication for a combined procedure was a diagnosed descending TAA and AAA with no significant risk factors for open aortic surgery or technical contraindications for endovascular treatment of TAA. RESULTS: One patient died in the peri-operative period while the other five patients all recovered well after surgery and were discharged with both aneurysms excluded. CONCLUSION: Endovascular treatment of TAA combined with a simultaneous open AAA repair is an efficient and relatively safe treatment modality in patients with TAA and AAA disqualified from endovascular repair. The fact that thoracotomy is not a necessity significantly lowers the complication rate in these patients. PMID- 15350572 TI - The changing incidence of femoral artery pseudoaneurysm. PMID- 15350573 TI - Pseudoaneurysm of the femoral artery in a HIV-infected man. PMID- 15350574 TI - Current treatment options for treating primary hyperhidrosis. PMID- 15350575 TI - Letter to the Editor, RE: EJVES 2004;28:104-107. PMID- 15350576 TI - Re: From Hippocrates to Palmaz-Schatz, the history of carotid surgery. PMID- 15350577 TI - Dynamic small animal lung imaging via a postacquisition respiratory gating technique using micro-cone beam computed tomography. AB - RATIONALE AND OBJECTIVES: Micro computed tomography is an important tool for small animal imaging. On many occasions, it is desirable to image lungs in a live instead of postmortem small animal to perform a pulmonary physiology study. Because the lungs are moving, gating with respect to the ventilatory phase has to be performed to reduce motion artifacts. Precapture ventilation gating may be difficult to achieve in some situations, which motivates us to propose and implement a simple postacquisition gating method. MATERIALS AND METHODS: Rats were used as the subjects in this study. A sequence of low-dose projection images were acquired at 30 frames per second for each view angle. During each capture sequence the rat undergoes multiple ventilation cycles. Using the sequence of projection images, an automated region of interest algorithm, based on integrated grayscale intensity, tracts the ventilatory phase of the lungs. In the processing of an image sequence, multiple projection images are identified at a particular phase and averaged to improve the signal-to-noise ratio. The resulting averaged projection images from different view angles are input to a Feldkamp cone-beam algorithm reconstruction algorithm to obtain isotropic image volumes. RESULTS: Reconstructions with reduced movement artifacts are obtained. In the gated reconstruction, registration of the bone is much better, the edge of the lung is clearly defined, and structures within the lung parenchyma are better resolved. Also, different phases of a breathing cycle can be reconstructed from one single tomographic scan by the proposed gating method. CONCLUSION: A postacquisition gating method using the phase information encoded in the 2-dimensional cone beam projections is proposed. This method is simple to implement and does not require additional experimental set-up to monitor the respiration. It may find applications in lung tumor detection, dynamic pulmonary physiology studies, and the respiratory systems modeling. Minimal motion artifact data sets improve qualitative and quantitative analysis techniques that are useful in physiologic studies of pulmonary structure and function. PMID- 15350578 TI - Impact of radiologic imaging on the surgical decision-making process in suspected appendicitis in children. AB - RATIONALE AND OBJECTIVES: To evaluate how the surgeons' decision-making process in appendicitis in children is affected by radiologic imaging. MATERIALS AND METHODS: Prospective study including 593 children with suspected appendicitis was conducted. The surgeon's initial clinical disposition was recorded, designating the patient for operation, observation, or discharge. Thereafter, the patients were randomized to undergo either ultrasound only or ultrasound and abdominal computed tomography. The studies were evaluated by radiologists, who indicated if appendicitis was present or not. After radiology was completed, the surgeon re examined the patient and made the final disposition. The change of disposition pathway was recorded. Final diagnoses were established by means of surgical, histopathologic, and/or clinical follow-up findings. RESULTS: Two hundred forty four patients had appendicitis. The initial clinical disposition called for 88 operations, 338 observations, and 167 discharges. In total, 347 patients had their treatment plan changed from the initial disposition, resulting in 252 operations, 65 observations, and 276 discharges. In 11 patients, an unnecessary operation was possibly avoided. In 28 patients who turned out to have appendicitis, a possible inappropriate discharge was avoided. Eighteen patients had a false-negative radiologic diagnosis. Of these, 17 underwent surgery because of convincing clinical findings. The difference between the impact on surgeons' decision-making between the two randomized groups was not substantially different. The negative appendectomy rate was 3.7%. CONCLUSION: Radiologic imaging with ultrasound and/or computed tomography provides valuable guidance whether a patient should be discharged, observed, or given surgical treatment, leading to beneficial changes in management plan. Still, false-negative results may occur and a close clinical re-examination is of utmost importance for the appropriate final decision. PMID- 15350579 TI - Multireader, multicase receiver operating characteristic analysis: an empirical comparison of five methods. AB - RATIONALE AND OBJECTIVES: Several statistical methods have been developed for analyzing multireader, multicase (MRMC) receiver operating characteristic (ROC) studies. The objective of this article is to increase awareness of these methods and determine if their results are concordant for published datasets. MATERIALS AND METHODS: Data from three previously published studies were reanalyzed using five MRMC methods. For each method the 95% confidence intervals (CIs) for the mean of the readers' ROC areas for each diagnostic test, the P value for the comparison of the diagnostic tests' mean accuracies, and the 95% CIs for the mean difference in ROC areas of the diagnostic tests were reported. RESULTS: Important differences in P values and CIs were seen when using parametric versus nonparametric estimates of accuracy, and there were the expected differences for random-reader versus fixed-reader models. Controlling for these differences, the Dorfman-Berbaum-Metz (DBM), Obuchowski-Rockette, Beiden-Wagner-Campbell, and Song's multivariate Wilcoxon-Mann-Whitney (WMW) methods gave almost identical results for the fixed-reader model. For the random-reader model, the DBM, Obuchowski-Rockette, and Beiden-Wagner-Campbell methods yielded approximately the same inferences, but the CIs for the Beiden-Wagner-Campbell method tend to be broader. Ishwaran's hierarchical ROC sometimes yielded significance not found with other methods. Song's modification of DBM's jack-knifing algorithm sometimes led to different conclusions than the original DBM algorithm. CONCLUSION: In choosing and applying MRMC methods, it is important to recognize: (1) the distinction between random-reader and fixed-reader models, the uncertainties accounted for by each, and thus the level of generalizeability expected from each; (2) assumptions made by the various MRMC methods; and (3) limitations of a five- or six-reader study when the reader variability is great. PMID- 15350580 TI - Iron oxide nanoparticles as magnetic resonance contrast agent for tumor imaging via folate receptor-targeted delivery. AB - RATIONALE AND OBJECTIVE: Targeted delivery is a highly desirable strategy for diagnostic imaging because of enhanced efficacy and reduced dosage/toxicity. Receptor-targeting was used to deliver contrast-producing superparamagnetic iron oxide (IO) nanoparticle to receptor-expressing tumors for in vivo magnetic resonance (MR) imaging. MATERIALS AND METHODS: Nanometer-sized, dextran-coated (maghemite) IO particles were prepared by a precipitation method. They were tethered with N-hydroxysuccinimide-folate and fluorescence isothiocyanate (FITC). For in vitro study of delivery specificity and efficiency, KB cells, a human nasopharyngeal epidermal carcinoma cell line expressing surface receptors for folic acid, were used as positive targets, and A549 cells, a human lung carcinoma cell line which lacks folate receptors, were used as negative control targets. In vivo MR images were obtained using mouse models with subcutaneous tumor xenografts grown from implanted KB cells. RESULTS: Internalization of nanoparticles into targeted cells only occurred when IO was conjugated to folate and when the folate receptors are available and accessible on the cells. The endocytosis was efficient and rapid, as 97.5% KB cells cultured with folate-FITC IO showed FITC uptake after 1 hour of incubation. In in vivo MR imaging, an average intensity decrease of 38% was observed from precontrast to postcontrast images of the tumor, which was about three times the intensity decrease observed at a non-tumor-bearing muscle. CONCLUSION: Successful in vivo MR imaging of folate receptor-expressing tumors targeted by IO nanoparticles was demonstrated for the first time. PMID- 15350581 TI - The effects of geometric distortion correction on motion realignment in fMRI. AB - RATIONALE AND OBJECTIVES: Subject motion is well recognized as a significant impediment to resolution and sensitivity in functional magnetic resonance imaging (fMRI). A parallel confounder to fMRI data quality is geometric image distortion, particularly at high field strengths, due to susceptibility-induced magnetic field inhomogeneity. Consequently, many high-field echo-planar imaging methods incorporate a post-processing distortion correction by acquiring a field map of the sample prior to the fMRI measurement. However, field mapping methods impose a spatial mask on the data, since field information is only obtainable from regions with adequate signal-to-noise ratio (SNR). This masking, when applied to subsequent images in the fMRI time series, can clip the effects of motion, resulting in inaccurate estimation and correction of motion-based changes in the images. MATERIALS AND METHODS: The effects of geometric distortion correction on automated realignment (motion correction) of fMRI data are investigated from data acquired at 4 T. The results of image realignment with and without prior application of distortion correction are compared, using the estimated motion parameters and overall image realignment as metrics. RESULTS: The application of field-map-based distortion correction prior to image realignment reduces the amount of motion detected by a standard motion correction algorithm. Moreover, motion correction applied before distortion correction is shown to result in superior realignment of motion-correction images. CONCLUSION: It is preferable to perform motion realignment prior to correcting for geometric distortion. PMID- 15350582 TI - Automated lung segmentation for thoracic CT impact on computer-aided diagnosis. AB - RATIONALE AND OBJECTIVES: Automated lung segmentation in thoracic computed tomography scans is essential for the development of computer-aided diagnostic (CAD) methods. A core segmentation method may be developed for general application; however, modifications may be required for specific clinical tasks. MATERIALS AND METHODS: An automated lung segmentation method has been applied (1) as preprocessing for automated lung nodule detection and (2) as the foundation for computer-assisted measurements of pleural mesothelioma tumor thickness. The core method uses gray-level thresholding to segment the lungs within each computed tomography section. The segmentation is revised through separation of right and left lungs along the anterior junction line, elimination of the trachea and main bronchi from the lung segmentation regions, and suppression of the diaphragm. Segmentation modifications required for nodule detection include a rolling ball algorithm to include juxtapleural nodules and morphologic erosion to eliminate partial volume pixels at the boundary of the segmentation regions. RESULTS: For automated lung nodule detection, 4 of 82 actual nodules (4.9%) were excluded from the lung segmentation regions when the core segmentation method was modified compared with 14 nodules (17.1%) excluded without modifications. The computer-assisted quantification of mesothelioma method achieved a correlation coefficient of 0.990 with 134 manual measurements when the core segmentation method was used alone; correlation was reduced to 0.977 when the segmentation modifications, as adapted for the lung nodule detection task, were applied to the mesothelioma measurement task. CONCLUSION: Different CAD applications impose different requirements on the automated lung segmentation process. The specific approach to lung segmentation must be adapted to the particular CAD task. PMID- 15350583 TI - In vivo imaging of MLC2v-luciferase, a cardiac-specific reporter gene expression in mice. AB - RATIONALE AND OBJECTIVES: A reporter or marker gene that is detectable by in vivo imaging permits longitudinal monitoring of certain fundamental biological processes (eg, differentiation) within the context of physiologically authentic environments. Tissue-specific expression of a reporter gene can be achieved when it is under the transcriptional control of a tissue-specific promoter. The objective of this study was to construct a plasmid vector containing firefly luciferase (Fluc) marker gene downstream of the promoter sequence of rat ventricular myosin light chain 2 (MLC2v); to detect the in vivo expression of this cardiac-specific reporter (MLC2v-Fluc) in the mouse heart by bioluminescent imaging; and to correlate the bioluminescent signal with postmortem luminometer assay. MATERIALS AND METHODS: MLC2v-Fluc plasmid was generated by molecular cloning of 3 kb promoter sequence into a pGL3-Basic vector containing the Fluc reporter. Twenty microg of MLC2v-Fluc plasmid DNA in phosphate-buffered saline was directly injected into mouse myocardium through a midline sternotomy. RESULTS: At 1 week after injection, MLC2v-Fluc expression was detected by in vivo bioluminescent imaging in 60% of injected animals; the average in vivo signal intensity was (1.5 +/- 0.6) x 10(4) radiance (p/sec/cm2/sr); in vivo signal was well above the detection threshold over 3 weeks after injection. In vivo bioluminescent signal is correlated (r2 = 0.8) with the luminometer assay results from homogenized heart samples. CONCLUSION: The capability of noninvasive imaging of the MLC2v-Fluc in the heart will encourage applications that aim at monitoring and tracking the marker gene expression over time in cells undergoing cardiac differentiation. PMID- 15350584 TI - A mouse optical simulation environment (MOSE) to investigate bioluminescent phenomena in the living mouse with the Monte Carlo method. AB - RATIONALE AND OBJECTIVES: As an important part of bioluminescence tomography, which is a newly developed optical imaging modality, mouse optical simulation environment (MOSE) is developed to simulate bioluminescent phenomena in the living mouse and to predict bioluminescent signals detectable outside the mouse. This simulator is dedicated to small animal optical imaging based on bioluminescence. MATERIALS AND METHODS: With the parameters of biological tissues, bioluminescent sources, and charge coupled device (CCD) detectors, the 2 dimensional/3-dimensional MOSE simulates the whole process of the light propagation in 2-dimensional/3-dimensional biological tissues using the Monte Carlo method. Both the implementation details and the software architecture are described in this article. RESULTS: The software system is implemented in the Visual C++ programming language with the OpenGL techniques and has a user friendly interface facilitating interactions relevant to bioluminescent imaging. The accuracy of the system is verified by comparing the MOSE results with independent data from analytic solutions and commercial software. CONCLUSION: As shown in our simulation and analysis, the MOSE is accurate, flexible, and efficient to simulate the photon propagation for bioluminescence tomography. With graduate refinements and enhancements, it is hoped that the MOSE will become a standard tool for bioluminescence tomography. PMID- 15350585 TI - Observation of microvasculatures in athymic nude rat transplanted tumor using synchrotron radiation microangiography system. AB - RATIONALE AND OBJECTIVES: The diagnostic potential of tumor microvasculatures with the monochromatic synchrotron radiation microangiography (MSRA) system was examined in an experimental model using athymic nude rats. MATERIALS AND METHODS: In the lower abdominal wall in athymic nude rats (male, 120-150 g, 6 weeks old), 1 x 10(7) cells of N-nitrosomethylurea-induced rat mammary adenocarcinoma was transplanted to prepare a lower abdominal wall tumor transplanted model. A microcatheter was inserted in the abdominal aorta in a rat under anesthesia, and microangiographic images of the normal inferior epigastric artery and vein were obtained using an MSRA system. Changes in the tumor microvasculatures were observed using an MSRA system on the day 7 (n = 3), day 14 (n = 3), day 21 (n = 3), and day 28 (n = 3) after transplant. In addition, we measured the microvessel density (MVD) with a computer using the binarization method. These results were compared among the histologic MVDs. RESULTS: The microvasculatures of tumors measuring 20-30 microm were observable by the MSRA system in the transplanted tumor model using the inferior epigastric artery and vein as the feeding vessels. The tumor microvasculatures were observed for 7-28 days after transplantation. The tumor MVD obtained by the binarization method increased as the tumor volume increased. When histologic MVD was compared with the fixed quantitative results of angiographic MVD, a significant correlation (r = 0.933, P <.01) was observed. CONCLUSION: These preliminary investigations indicate that MSRA proved to be suitable for quantification and appears to be a simple method for clearly imaging tumor microvasculatures. PMID- 15350586 TI - A primer on molecular biology for imagers: VII. Molecular imaging probes. PMID- 15350587 TI - Postoperative evaluation of complex aortovisceral and aortorenal reconstructions by magnetic resonance angiography. AB - RATIONALE AND OBJECTIVES: To assess the ability of magnetic resonance angiography (MRA) to evaluate complex vascular bypass reconstructions of the abdominal aorta and its major branches in the postoperative period. MATERIALS AND METHODS: Thirteen patients with bypass grafts connecting the aorta to visceral, renal, and lower limb inflow vessels were evaluated with MRA. Three of these patients were also studied with digital subtraction angiography soon after MRA was completed. MRA was evaluated for its ability to detect the grafts and to determine the degree of stenosis in the graft conduit or at the anastomoses to native vessels. RESULTS: Detection of graft conduits and anastomotic sites by MRA was 100% and 99%, respectively. Comparison with digital subtraction angiography in a subset of the patients showed a 100% agreement between the two modalities in their description of stenotic disease in graft conduits and 95% agreement in stenosis characterization at graft anastomotic sites. CONCLUSION: MRA of complex aortic reconstructions with bypass grafts to its major abdominal branches arteries accurately describes the resulting complicated vascular anatomy and likely has a high degree of correlation to digital subtraction angiography in describing the disease within the bypass grafts. PMID- 15350588 TI - Blended learning in medical education: use of an integrated approach with web based small group modules and didactic instruction for teaching radiologic anatomy. AB - RATIONALE AND OBJECTIVES: To describe the development of and assess student satisfaction with a blended learning method for teaching radiologic anatomy that integrates web-based instruction with small group and didactic teaching. MATERIALS AND METHODS: In 2002 the teaching of radiologic anatomy to first-year medical students was changed from group learning (20-30 students with a preceptor and films at a viewbox) to a blended learning model that included a brief didactic introduction followed by small group (7-8 students) web-based structured learning modules with rotating lab instructors. In 2003 the modules were changed to include self-study cases prior to the lab, follow-up cases, and twice-weekly optional review sessions. Students and lab instructors were surveyed for their response to the content and design of the sessions. RESULTS: Course surveys in 2001, with a response rate of 84%, showed 58 negative comments regarding inconsistency between various instructors. Individual response rates for 2002 for radiologic anatomy teaching sessions (RadLab) surveys ranged from 56%-81%, dropping as the course progressed. All RadLabs were rated "very useful" or "useful," except the cardiovascular lab, which was not designed as an interactive module. In 2003, after redesign of the cardiovascular lab in the same format as the other labs, all RadLabs were rated 2.4 or better (useful). CONCLUSION: An integration of computers with small and large group didactic instruction allow optimal use of faculty, conform to accepted theories of adult learning, and are well-accepted by students. PMID- 15350589 TI - Evaluation of an online faculty appraisal instrument: comparison of resident and faculty perceptions. AB - RATIONALE AND OBJECTIVES: Our purpose was to compare faculty and resident perceptions regarding an online faculty appraisal instrument and the potential impact of the appraisal process. MATERIALS AND METHODS: Faculty members and residents at an academic medical center diagnostic radiology department were asked to complete anonymous surveys that sought feedback regarding an online faculty appraisal form and process. Questions were asked regarding the relative importance of various faculty performance measures, preferences for narrative versus quantitative components of the instrument, and the likely impact of the evaluation process on future faculty behavior. RESULTS: The survey was completed by 19 (45%) of 42 faculty members and by 16 (80%) of 20 residents who participated in the initial faculty appraisal process. Residents and faculty both assigned generally high rankings to the six measures of faculty performance. The vast majority (86 %) of faculty and residents found the narrative component of the survey (either alone or in combination with quantitative data) most helpful. Based on the appraisal process, 74% of the faculty planned to modify one or two aspects of their behavior, whereas a significant minority (44%) of residents thought that the faculty would not make any changes. CONCLUSION: Faculty and residents generally agree on a core set of faculty performance measures, and both groups show a preference for an appraisal instrument that incorporates a narrative component. Concerning the perceived impact of the appraisal process, faculty members were slightly more optimistic than residents regarding its ability to serve as an impetus for faculty behavioral changes. PMID- 15350590 TI - Phallacidin stains the mitotic spindle of the diatom Pinnularia spp. AB - Mitotic spindles of the diatom Pinnularia viritiformis stained with the fluorescent actin-labelling reagent bodipyphallacidin revealed actin among the chromosomes and extending along the spindle to the poles. This is the first report of actin's presence within spindles of this ecologically important group of organisms. Since diatom mitoses have a number of marked differences compared to that of many other eukaryotes (Int Rev Cytol 128 (1991) 63; Cell 14 (1978) 455), the present observations substantially extend the diversity of mitotic spindle types in which there is evidence of spindle actin. PMID- 15350591 TI - Urokinase plasminogen activator (uPA) is a positive regulator of outer root sheath keratinocyte proliferation. AB - Urokinase plasminogen activator (uPA), a serine proteinase, is important in the development and epidermal wound healing, and seems to play a regulatory role in the proliferation of mouse epidermal keratinocytes (KC). In the present study, we found detectable uPA expression in outer root sheath (ORS) KC in the early anagen phase in mouse vibrissa follicles, but not in the late anagen or in the telogen and categen phases. uPA was also detected in ORS KC cultured from neonatal mice vibrissa. Specific exogenous inhibitors of uPA, amiloride and uPA antibody, significantly reduced the proliferation of ORS KC. Thus uPA is consistently elevated in the hyperproliferative hair follicle KC, and inhibition of the enzyme decreases hair follicle KC proliferation. We deduce that uPA is a very important mediator of the hair follicle cycle because its activity correlates with ORS KC proliferation. PMID- 15350592 TI - Nitric oxide spatial distribution in single cultured hippocampus neurons: investigation by projection of reconstructed 3-D image and visualization technique. AB - Recent studies have revealed a non-homogeneous distribution of nitric oxide synthase (NOS) in neurons. However, it is not yet clear whether the intracellular distribution of NOS represents the intracellular nitric oxide (NO) distribution. In the present study, software developed in our laboratory was applied to the reconstructed image obtained from confocal slice images in order to project the 3 D reconstructed images in any direction and to cut the neuron in different sections. This enabled the spatial distribution of NO to be visualized in any direction and section. In single neurons, NO distribution was seen to be heterogeneous. After stimulation with glutamate, the spatial changes in different areas of the neuron were different. These findings are consistent with immunocytochemical data on the intracellular localization of nNOS in hippocampus neurons, and will help to elucidate the specificity of nitric oxide signaling. Finally, the administration of SNAP and L-NAME was used to examine DAF-2 distribution in the neurons. The results showed this distribution to be homogenous; therefore, it did not account for the NO distribution results. PMID- 15350593 TI - Prooxidative effects of TEMPO on human erythrocytes. AB - The prooxidative effects of 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) were observed in human erythrocytes. Incubation of red blood cells with the membrane permeable TEMPO leads to a decrease in the concentration of intracellular reduced glutathione, accompanied by the reduction of TEMPO. Extracellular ferricyanide inhibited the loss of glutathione and reduction of TEMPO. TEMPO induced glutathione release from the cells and oxidation of hemoglobin to methemoglobin; ferricyanide prevented these effects. These results indicate that TEMPO may act as an oxidant to erythrocytes, whilst extracellular ferricyanide protects against its effects. PMID- 15350594 TI - Impact of hypotonic solutions on the stromal swelling, lactate dehydrogenase and aldehyde dehydrogenase activity of the keratocytes of the bovine cornea. AB - The present studies were designed to explore the relationship between the swelling-related changes of the collagen-cell (keratocyte) matrix of the corneal stroma, and the integrity of the cells. From recent postmortem eyes of adult cattle, complete stroma preparations were dissected out and allowed to swell in solution (free swelling) or enclosed within a 12 kDa cut-off dialysis membrane with or without spacers. The swelling was at 4 degrees C with either water, a hypotonic phosphate-buffered saline (PBS, pH 7.0), a hypotonic mixed salt (MS) solution (pH 7.5), or an isotonic mixed salt solution with glucose (pH 7.5). Measures of tissue wet mass and thickness and analyses of the soluble protein, LDH and ALDH activity in the solutions were made. The relative swelling of the stroma preparations was greatest in water (to 624% of the original wet mass) > dilute PBS (to 404%) > dilute MS (to 381%) > MS with glucose (to 356%). The relative swelling was in the same order, but slightly less if the stroma preparations were enclosed in a dialysis tube with spacers, and substantially reduced when enclosed in a dialysis bag without spacers. With the use of hypotonic solutions, substantial quantities of proteinaceous material and enzyme activity were lost from the preparations, with the loss being proportional to the extent of swelling (p < 0.001). Swelling of an isolated corneal stroma, especially in hypotonic solutions, is associated with substantial loss of soluble protein and cytoplasmic enzyme activities, and so these solutions must be considered as cytotoxic to the keratocytes. PMID- 15350595 TI - Cloning and functional characterization of a rat Daxx that functions as a corepressor for the androgen receptor. AB - The androgen receptor (AR), a ligand-activated nuclear transcription factor, plays an important role in endocrine system development and homeostasis. AR mediated gene transcription is modulated by a wide variety of coregulator proteins, but corepressors of AR have not been well characterized. We have isolated a rat homologue of Daxx, a possible AR repressor. The clone obtained comprised a 2196 bp open reading frame, corresponding to a sequence of 731 amino acids with 80.5% homology to mouse. Basal transcription of the isolated clone was repressed in transient transcription experiments using a Gal4 reporter gene assay. Furthermore, a mammalian two-hybrid assay showed a strong interaction between Daxx and promyelocytic leukemia (PML) protein. These results indicate that the isolated clone is a rat homologue of Daxx. The rat Daxx (rDaxx) interacted with the androgen receptor (AR): in co-immunoprecipitation experiments, AR formed complexes with anti-Daxx antibody. Furthermore, overexpression of Daxx suppressed AR-mediated transcriptional activity in HeLa and CV1 cells in transient transfection experiments. Taken together, our results suggest that Daxx may function as a corepressor for the androgen receptor. PMID- 15350596 TI - Role of caveolin-1 and cytoskeletal proteins, actin and vimentin, in adipogenesis of bovine intramuscular preadipocyte cells. AB - We investigated the involvement of caveolin-1 and the cytoskeletal proteins, actin and vimentin, in the adipogenesis of bovine intramuscular preadipocyte (BIP) cells. Immunoblot analysis demonstrated that levels of caveolin-1 and actin gradually increased during adipose conversion in BIP cells, whereas a slight decrease was observed for vimentin. We found that part of the vimentin was clearly distributed to caveolin-1-enriched membrane fractions in BIP cells, but actin was not. During adipogenesis of BIP cells, treatment with the tubulin depolymerizer, nocodazole, significantly increased intracellular triglyceride accumulation compared to non-treated cells. Immunocytochemical analysis showed that actin microfilaments were significantly disrupted in nocodazole-treated cells. Also, a decrease in the localization of vimentin in caveolin-1-enriched fractions and a failure of vimentin to co-immunoisolate with caveolin-1 were observed in nocodazole-treated cells. These results suggest that a rearrangement of cytoskeletal proteins has a role in the intracellular accumulation of lipid droplets during adipogenesis of BIP cells. PMID- 15350597 TI - Karyotype analyses and studies on the nuclear DNA content in 30 genotypes of potato (Solanum tuberosum) L. AB - The cytophotometric estimation of 4C DNA content, and karyotypic and somatic chromosome number analyses were carried out in 30 genotypes comprising seven cultivars and 23 advanced breeding lines of Solanum tuberosum. Detailed karyotype analysis revealed genotype specific chromosomal characteristics and structural alterations in chromosomes of the genome, with a rare phenomenon of aneusomatic (2n = 4x + 2 = 50) condition in cv.K. Chandramukhi. The origin of this variation could be attributed to mitotic non-disjunction in the shoots giving rise to aneusomatic roots. Highly significant variations in the genome length, volume and total form percentage were noted at the cultivar level. The total chromosome length varied from 84.56 microm in cv.K. Pukhraj to 127.62 microm in MS/89-60, with an average value of 100.94 microm +/- 1.82. Total chromosome volume varied from 57.22 microm3 in MS/92-1090 to 132.64 microm3 in JW-160. Significant variations in the 4C DNA content (7.28-15.83 pg) were recorded at the cultivar level, with an exceptionally high DNA content (22.24 pg) in cv.K. Chandramukhi. This could be due to the aneusomatic condition of this genotype. Correlation studies revealed interdependence between the chromosomal and nuclear parameters of the genotypes. Structural alterations in the chromosomes, as well as loss or addition of highly repetitive sequences in the genome, caused variations in DNA content at the cultivar level. Variations in genomic structure and nuclear DNA content of the 48-chromosome genotypes suggest a genetic drift during microevolution, leading to the development of new cultivars. PMID- 15350598 TI - Expression of polypeptide GalNAc-transferases in hematopoietic stem/progenitor cells. AB - The members of the UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase (pp GalNAc-T) family, which transfer GalNAc to polypeptide serine and threonine residues, initiate mucin-type O-linked glycosylation. There are at least 13 functionally characterized members of this family in humans, but no studies have been reported of pp-GalNAc-T isoforms in hematopoietic cells. We isolated and purified CD34+ hematopoietic cells from adult bone marrow by magnetic cell sorting and induced them to differentiate into megakaryocytic lineage cells using an optimal combination of hematopoietic growth factors in serum-free liquid medium. RT-PCR revealed that CD34+ cells expressed pp-GalNAc-T1, T2, T3, T4, T6, T7, T10, T11 and T14, but not pp-GalNAc-T8, T9, T12 and T13. The megakaryocytic lineage cells showed significant increases in the expression of pp-GalNAc-T3, T8, T9, T10 and T13, but pp-GalNAc-T11 and T14 became undetectable. In summary, many pp-GalNAc-T isoforms were expressed in CD34+ cells but the expression pattern changed during differentiation into megakaryocytes. The expression patterns of pp GalNAc-Ts may be necessary to ensure proper O-glycosylation of mucin-type proteins expressed in CD34+ and megakaryocytic cells. PMID- 15350599 TI - Senescent phenotype achieved in vitro is indistinguishable, with the exception of Bcl-2 content, from that attained during the in vivo aging process. AB - Senescent phenotype can be attained by diverse agents, thus suggesting that there might be molecular differences between the senescence achieved in vivo and the senescence-like state attained in vitro under culture conditions. In this study we compare the senescent phenotype reached by cells derived from young animals when cultured in vitro with the one associated with the in vivo aging process. Several in vitro senescence parameters, including MTT reduction, proliferation rate, DNA synthesis, SA-beta-gal staining, and both in vivo and in vitro Bcl-2 content, were determined. Alterations in DNA electrophoretic mobility were evaluated to test differences in bulk chromatin structure. Our results indicate that although it is possible to achieve a senescent phenotype with cells derived from young animals aged in culture, this phenotype differs from the one observed in older animals, due to lack of in vivo damage inducers to which cells are being exposed during natural aging. PMID- 15350600 TI - Cortisol resistance in the New World revisited. AB - Insights into the molecular basis of glucocorticoid action have been obtained from the analysis of cortisol resistance. The glucocorticoid receptor (GR) in both New World primates and guinea pigs has a decreased affinity, in vivo, for cortisol; this is achieved by two distinct mechanisms. In the New World primates recent studies have identified a key role for co-chaperones. The amino acids responsible for cortisol resistance in the guinea pig GR lie not in the ligand binding pocket but on the surface of the receptor. We hypothesize that this region might be the site of interaction between the co-chaperones and the GR, and hence that the resistance occurs through the same mechanism, albeit from opposite sides. PMID- 15350601 TI - Chemotherapy targeted to cancers through tumoral hormone receptors. AB - Work on cytotoxic analogs of luteinizing hormone-releasing hormone (LH-RH), somatostatin and bombesin, designed for targeting chemotherapy to peptide receptors on various cancers, is reviewed here as the project is at advanced stages of development and clinical trials are pending. Cytotoxic analogs of LH RH, AN-152 and AN-207, containing doxorubicin (DOX) or 2-pyrrolino-DOX (AN-201), respectively, target LH-RH receptors and can be used for the treatment of prostatic, breast, ovarian and endometrial cancers and melanomas. AN-201 was also incorporated into the cytotoxic analog of somatostatin, AN-238, which can be targeted to receptors for somatostatin in prostatic, renal, mammary, ovarian, gastric, colorectal and pancreatic cancers as well as glioblastomas and lung cancers, suppressing the growth of these tumors and their metastases. A cytotoxic analog of bombesin AN-215, containing 2-pyrrolino-DOX, was likewise synthesized and successfully tested in experimental models of prostate cancer, small cell lung carcinoma, gastrointestinal cancers and brain tumors expressing receptors for bombesin/gastrin-releasing peptide. This new class of targeted cytotoxic peptide analogs might provide a more effective therapy for various cancers. PMID- 15350602 TI - P450 oxidoreductase deficiency: a new disorder of steroidogenesis with multiple clinical manifestations. AB - Combined partial deficiency of 17alpha-hydroxylase and 21-hydroxylase is well described, but patients' genes for these enzymes lack mutations. Recent work has identified mutations in the gene for P450 oxidoreductase (POR) in such patients. POR-deficient individuals have a broad range of disorders, from infants with congenital malformations to women with the polycysic ovary syndrome. POR transfers electrons to all microsomal P450 enzymes: its deficiency affects steroidogenesis, drug metabolism and other processes. PMID- 15350603 TI - Steroid receptors in human breast cancer. AB - Ovarian steroids, acting through nuclear receptors, are crucial players in normal breast development and cancer. Estrogen, in particular, is the focus of breast cancer therapies because tumours are often dependent on this steroid for growth. Recently, novel genes and/or protein isoforms of receptors for both estrogen and progesterone have been discovered, leading us to reappraise their roles in breast development and cancer. Recognition of changes in estrogen receptor biology that occur in the transition from normal development to cancer has emphasized its contribution to tumorigenesis. In addition, complex interactions with other signalling pathways, particularly growth factor pathways, have recently come to the forefront. These interactions might explain resistance to endocrine treatments and offer solutions in terms of novel therapeutic targets. PMID- 15350604 TI - Genomic and non-genomic interactions of PPARalpha with xenobiotic-metabolizing enzymes. AB - The hypolipidemic properties of fibrates, synthetic activators of the nuclear receptor, peroxisome proliferator-activated receptor alpha (PPARalpha), have been studied extensively. Recent observations indicate, however, that PPARalpha also functions as a regulator of endobiotic and xenobiotic metabolism in rodents and humans. Activators of PPARalpha affect xenobiotic-metabolizing enzymes (XMEs) at different levels. At the genomic level, the expression of numerous cytochrome P450 (CYP) and phase II conjugating genes is altered in a species-distinct manner on treatment with PPARalpha activators. As a result of such regulatory processes, PPARalpha affects the homeostasis of both its own natural ligands and other compounds including bile acids. At the non-genomic level, PPARalpha activators can act as competitive inhibitors for inactivating other molecules, leading to drug-drug interactions. These global effects of PPARalpha activators on the activity of XMEs are of physiological and pharmaceutical importance, and demonstrate that thorough studies of the actions on XMEs of each novel PPARalpha agonist are warranted. PMID- 15350605 TI - Genomics view of gonadotrope signaling circuits. AB - Gonadotropin-releasing hormone (GnRH) binds to the pituitary GnRH receptor to activate signal transduction cascades that ultimately modulate gonadotropin biosynthesis. Comprehensive studies of the GnRH-activated gene program in the LbetaT2 gonadotrope cell line have greatly increased our knowledge of the number of early and intermediate gene transcripts that are modulated by GnRH. Among the classes of gene induced are several whose protein products provide feedback to various levels of signaling pathways, suggesting that gene induction forms an integral component of signal transduction and contributes to longer-timescale feedback and feedforward loops. High-throughput quantitative genomic studies, mathematical modeling and biochemical studies are beginning to delineate the organization and function of the signal-decoding and logic circuit modules of the gonadotrope's signal transduction network. PMID- 15350606 TI - ATRX and sex differentiation. AB - Most interest in the gene encoding the alpha-thalassemia, mental retardation, X linked protein (ATRX) has traditionally been focused on its role in brain development and globin regulation. However, mutations in the ATRX gene also cause varying degrees of gonadal and urogenital abnormalities. These range from small testes to ambiguous external genitalia in XY individuals. ATRX is of great interest because this very large protein is one of the least understood proteins involved in mammalian sexual development. Although the biochemical role of ATRX remains unknown, recent and emerging evidence implicates ATRX in chromatin remodeling. We review what is known about ATRX as a chromatin remodeling protein, and its role in mammalian sex differentiation. PMID- 15350607 TI - Lactate and energy metabolism in male germ cells. AB - Various alterations in germ cell proliferation/differentiation, survival and energy metabolism are potentially involved in hypospermatogenesis leading to male infertility. Several reviews have been devoted to the different processes whose alteration might underlie hypospermatogenesis, except for energy metabolism in the testis. Energy metabolism in the testis exhibits some specificity in that lactate is the central energy metabolite used by germ cells. This metabolite is produced by somatic Sertoli cells, transported and used by germ cells in the context of an active cooperation under the control of the endocrine system and local cytokines. In this review, we present and discuss relevant published data on energy metabolism in male germ cells with a specific emphasis on lactate. PMID- 15350608 TI - period expression in the honey bee brain is developmentally regulated and not affected by light, flight experience, or colony type. AB - Changes in circadian rhythms of behavior are related to age-based division of labor in honey bee colonies. The expression of the clock gene period (per) in the bee brain is associated with age-related changes in circadian rhythms of behavior, but previous efforts to firmly associate per brain expression with division of labor or age have produced variable results. We explored whether this variability was due to differences in light and flight experience, which vary with division of labor, or differences in colony environment, which are known to affect honey bee behavioral development. Our results support the hypothesis that per mRNA expression in the bee brain is developmentally regulated. One-day-old bees had the lowest levels of expression and rarely showed evidence of diurnal fluctuation, while foragers and forager-age bees (> 21 days of age) always had high levels of brain per and strong and consistent diurnal patterns. Results from laboratory and field experiments do not support the hypothesis that light, flight experience, and colony type influence per expression. Our results suggest that the rate of developmental elevation in per expression is influenced by factors other than the ones studied in our experiments, and that young bees are more sensitive to these factors than foragers. PMID- 15350609 TI - Plasmodium berghei ookinetes induce nitric oxide production in Anopheles pseudopunctipennis midguts cultured in vitro. AB - The Anopheles pseudopunctipennis nitric oxide synthase gene (ApNOS) was identified and its partial sequence showed high homology with NOS from A. stephensi, A. gambiae (putative sequence), and Drosophila melanogaster. ApNOS was mainly expressed in male and female adult mosquitoes and was induced by a blood meal. Nitric oxide (NO) was produced by in vitro-cultured mosquito midguts inoculated by enema with Plasmodium berghei ookinetes, Saccharomyces cerevisiae, Gram-positive bacteria (Micrococcus luteus), but not with Gram-negative bacteria (Klebsiella pneumoniae, Escherichia coli or Serratia marcescens). Dihydroxyphenylalanine (L-DOPA) oxidation induced the generation of NO in midguts in vitro, and hydrogen peroxide generated during its oxidation induced ApNOS expression. P. berghei ookinetes exposed in vitro to L-DOPA and sodium nitroprusside (a NO generator) were killed. These observations demonstrate that reactive oxygen and nitrogen intermediates constitute a part of the cytotoxic arsenal employed by Anopheles mosquitoes against microbial pathogens and Plasmodium ookinetes. PMID- 15350610 TI - A diverse family of serine proteinase genes expressed in cotton boll weevil (Anthonomus grandis): implications for the design of pest-resistant transgenic cotton plants. AB - Fourteen different cDNA fragments encoding serine proteinases were isolated by reverse transcription-PCR from cotton boll weevil (Anthonomus grandis) larvae. A large diversity between the sequences was observed, with a mean pairwise identity of 22% in the amino acid sequence. The cDNAs encompassed 11 trypsin-like sequences classifiable into three families and three chymotrypsin-like sequences belonging to a single family. Using a combination of 5' and 3' RACE, the full length sequence was obtained for five of the cDNAs, named Agser2, Agser5, Agser6, Agser10 and Agser21. The encoded proteins included amino acid sequence motifs of serine proteinase active sites, conserved cysteine residues, and both zymogen activation and signal peptides. Southern blotting analysis suggested that one or two copies of these serine proteinase genes exist in the A. grandis genome. Northern blotting analysis of Agser2 and Agser5 showed that for both genes, expression is induced upon feeding and is concentrated in the gut of larvae and adult insects. Reverse northern analysis of the 14 cDNA fragments showed that only two trypsin-like and two chymotrypsin-like were expressed at detectable levels. Under the effect of the serine proteinase inhibitors soybean Kunitz trypsin inhibitor and black-eyed pea trypsin/chymotrypsin inhibitor, expression of one of the trypsin-like sequences was upregulated while expression of the two chymotrypsin-like sequences was downregulated. PMID- 15350611 TI - Utilization of pre-existing energy stores of female Aedes aegypti mosquitoes during the first gonotrophic cycle. AB - Pre-existing energy reserves may play an important role in regulating the utilization of blood meal proteins in female anautogenous mosquitoes. Determining the fate of reserves derived from the sugar meal and larval food during the first gonotrophic cycle would help to elucidate the relative contributions of larval and adult nutrition to survival and reproduction. We measured the allocation of pre-blood-meal reserves to egg production or energy production during the first gonotrophic cycle by using [14C]-labeled female Aedes aegypti mosquitoes. Feeding adults [3,4-14C]-glucose labeled the glycogen and sugar stores (approximately 50%), lipid stores (approximately 25%), and protein and amino acid stores (approximately 25%). During the first gonotrophic cycle, about 60% of the glycogen and sugar stores were metabolized and all were used for energy production. About 33% of the labeled protein and 72% of the labeled amino acid stores were metabolized, with about 9% being transferred to the eggs and the rest oxidized. About 30% of the lipid was metabolized, with about 65% being transferred to the eggs and the rest oxidized. Feeding [1-14C]-oleic acid to larvae effectively labeled adult lipid stores with about 75% of the label in lipid stores and 16% in proteins and 6% in glycogen. During the first gonotrophic cycle, about 35% of the labeled lipid stores were metabolized, with equal amounts being oxidized and transferred to the eggs. None of the other maternal stores labeled by fatty acid were metabolized during the first gonotrophic cycle. These results show that carbohydrate reserves are a critical source of energy during the first gonotrophic cycle, while lipid reserves are used equally for energy production and provisioning the eggs. PMID- 15350612 TI - Identification of a new member of PBAN family and immunoreactivity in the central nervous system from Adoxophyes sp. (Lepidoptera: Tortricidae). AB - Production of sex pheromones, Z9-14:OAc and Z11-14:OAc, of the smaller tea tortrix, Adoxophyes sp. was stimulated by injection of the female or male head extracts as well as synthetic pheromone biosynthesis activating neuropeptide (PBAN) into decapitated females. The amount of pheromone produced reached a maximum level 3 h after injection of synthetic PBAN into females. A cDNA isolated from brain-suboesophageal ganglion complex (Br-SEG) of A. sp. females contained an ORF of 576 nucleotides encoding 192 amino acids. Based on endoproteolytic sites, it can be predicted to be cleaved into five putative peptide domains including PBAN and four other neuropeptides. Ado-PBAN consisting of 31-amino acids is the shortest PBAN so far reported. Four other putative PBAN-encoding gene neuropeptides (PGN) are predicted with PGN-24, PGN-7, PGN-20, and PGN-8 amino acids. All of the peptides are amidated in their C-termini with a FXPR(or I, K)L structure, except for PGN-8 (TVKLTPRLamide). PBAN-like immunoreactive material was observed in Br, SEG and ventral nerve cord (VNC) of the female adult. In the brain, 5-7 pairs of neurons containing PBAN-like immunoreactivity were found in each protocerebral hemisphere. Three groups of cell clusters found in the SEG corresponded to the mandibular, maxillary and labial neurons as in other moths. PBAN-like immunoreactive neurons in the VNC were found in thoracic (three pairs) and abdominal ganglia (two pairs). As compared to other moths, a relatively low similarity of peptide sequences deduced from Ado-PBAN gene and a different expression pattern of PBAN-like immunoreactivity could indicate phylogenetical distance from the other species. PMID- 15350613 TI - Differential gene expression between alate and dealate queens in the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). AB - The transition of fire ant queens from alates to dealates, following a mating flight, is associated with numerous important physiological changes. A molecular analysis of gene expression differences that occur between alates and dealates was performed using the suppression subtractive hybridization (SSH) method. 983 SSH clones were arrayed and screened by dot blot hybridization, followed by Northern blot analysis for selected clones. Gene expression profiles throughout fire ant development were determined using semi-quantitative reverse transcriptase polymerase chain reactions (RT-PCR). The cytochrome c oxidase subunit II and STARS (striated muscle activator of Rho signaling) transcripts were expressed at higher levels in dealates compared to alates and may be involved in the programmed cell death of the flight muscles. Three different vitellogenin genes and two unique yellow g-like genes were identified that may be closely associated with the reproductive system and/or nutrient transport. Two putative antibacterial peptides, abaecin and hymenoptaecin precursors, were highly expressed in dealate queens, suggesting that they are present as an immune system component during this important stage of fire ant development. The genes identified in this study may be utilized as novel targets for fire ant control and will also provide molecular markers for studies of other social insects. PMID- 15350614 TI - Vitellogenin of the parasitoid wasp, Encarsia formosa (Hymenoptera: Aphelinidae): gene organization and differential use by members of the genus. AB - The vitellogenin (Vg) gene of the parasitoid wasp, Encarsia formosa (Hymenoptera: Aphelinidae), has been cloned and sequenced. The gene codes for a protein consisting of 1814 amino acids in seven exons. The position of the six introns in the E. formosa gene align with those inferred for the Vg gene of the honeybee, Apis mellifera. The position of two introns in the hymenopteran sequences are shared with every full-length insect Vg gene characterized to date. The deduced amino acid sequence of the E. formosa Vg gene most closely resembles that of the ichneumonid parasitoid, Pimpla nipponica (38% identity). The gene product, less the putative signal peptide, contains large quantities of serine (11.3% of total residues) but lacks the extensive polyserine tracts found in the Vgs of insects outside the apocritan Hymenoptera. The gene also codes for the highest level of lysine (9.5%), and lowest levels of phenylalanine (2.6%) and tyrosine (2.3%), observed in any insect Vg characterized to date. The mature gene product retains 12 cysteine residues in positions conserved in other insect Vgs. Ovary homogenates suggest that processed Vg is stored in the egg as an uncleaved molecule of approximately 200 kDa. Vg expression was examined in three additional Encarsia species. The protein was found in female E. sophia and E. luteola, but not in male E. luteola or female E. pergandiella. Despite extensive screening of a phage library prepared from E. pergandiella genomic DNA, a Vg gene was not detected in this species. PMID- 15350615 TI - cDNA cloning, gene structure, and expression of Broad-Complex (BR-C) genes in the silkworm, Bombyx mori. AB - To clarify the molecular mechanisms of metamorphosis, we analyzed the Broad Complex (BR-C) gene in the silkworm, Bombyx mori. We cloned cDNAs for the full coding regions of the Z1, Z2, and Z4 isoforms of BR-C. The Z3 zinc finger sequence was found in the 3'UTR of the Z2 isoform. The predicted amino acid sequence showed high homology with Drosophila and Manduca BR-C proteins. Five bacterial artificial chromosome (BAC) clones were screened from a Bombyx BAC library. Restriction enzyme cleavage maps of 170 kb regions were constructed, and a total of 25.8 kb were sequenced. The BAC analysis showed that the 5'UTR of the BR-C gene consists of the first two exons, while the coding region contains a core region domain with five exons and four zinc finger exons in the order Z1, Z4, Z2, and Z3. Expression analysis revealed 9.5, 6.5, and 5.5 kb BR-C transcripts. These increased during the spinning ecdysone peak on day 6 of the fifth instar when pupal commitment occurs in the Bombyx epidermis. In addition, a small amount of BR-C mRNA was detected in the epidermis before this peak. BR-C mRNA was also expressed in the fat body from day 1 in the fourth instar to day 7 in the fifth instar. PMID- 15350616 TI - Three recombinant serine proteinase inhibitors expressed from the coding region of the thrombin inhibitor dipetalogastin. AB - Dipetalogastin is a potent thrombin inhibitor from Dipetalogaster maximus. The cDNA of dipetalogastin codes for a large protein which consists of six Kazal-type domains. There are three tandem, homologous regions each including two domains. Three biologically active recombinant proteins rDI, rDII and rDIII each corresponding to one region of the dipetalogastin cDNA were expressed, purified and investigated with regard to their biological activities. rDI and rDII with molecular masses of 12,660 and 12,911 Da, respectively, proved to be potent thrombin inhibitors. The investigation of their influences on amidolytic activities of different serine proteases showed no inhibition of factor Xa (FXa) and alpha-chymotrypsin. At a large molar excess of rDI and rDII over the enzymes only low effects on the activities of trypsin and plasmin were observed. rDIII differs much from the both others. An inhibition of thrombin was found only at a molar excess of rDIII over the enzyme. Furthermore, an inhibition of trypsin and low effects on plasmin were detected at a molar excess of inhibitor over these enzymes. These results indicate that rDIII is active against thrombin, trypsin and plasmin, and finally possesses no specificity for only one serine proteinase. PMID- 15350617 TI - Dynamic of ligand binding to Drosophila melanogaster ecdysteroid receptor. AB - Ligand binding to ecdysone receptor (EcR) is an autonomous function of the ligand binding domain (LBD) and is not modified by other receptor domains or tags fused to the LBD. Association and dissociation velocity of hormone to EcR was studied in the absence and presence of its main dimerization partner Ultraspiracle (USP). Mutational analysis of the EcR(LBD) revealed that ligand entry and exit is affected differently by the same point mutation, indicating that different pathways are used for association and dissociation of the ligand. Heterodimerization with wild type USP(LBD) increases ligand association to EcR(LBD) about fivefold and reduces dissociation 18-fold. Opposite effects of the same mutation (N626K) on dissociation velocity of ligand in EcR and EcR/USP indicate that not only hormone binding itself, but also the kinetic behaviour of ligand binding is modified by the dimerization partner. A general effect of the point mutations on the 3D architecture seems unlikely due to the highly selective effects on the kinetics of hormone binding. PMID- 15350618 TI - Phantom encodes the 25-hydroxylase of Drosophila melanogaster and Bombyx mori: a P450 enzyme critical in ecdysone biosynthesis. AB - We have reported recently the identification and characterization of the last three mitochondrial cytochrome P450 enzymes (CYP) controlling the biosynthesis of 20-hydroxyecdysone, the molting hormone of insects. These are encoded by the following genes: disembodied (dib, Cyp302a1, the 22-hydroxylase); shadow (sad, Cyp315a1, the 2-hydroxylase); and shade (shd, Cyp314a1, the 20-hydroxylase). Employing similar gene identification and transfection techniques and subsequent biochemical analysis of the expressed enzymatic activity, we report the identity of the Drosophila gene phantom (phm), located at 17D1 of the X chromosome, as encoding the microsomal 25-hydroxylase (Cyp306a1). Similar analysis following differential display-based gene identification has also resulted in the characterization of the corresponding 25-hydroxylase gene in Bombyx mori. Confirmation of 2,22,25-trideoxyecdysone (3beta,5beta-ketodiol) conversion to 2,22-dideoxyecdysone (3beta,5beta-ketotriol) mediated by either Phm enzyme employed LC, MS and definitive NMR analysis. In situ developmental gene analysis, in addition to northern, western and RT-PCR techniques during Drosophila embryonic, larval and adult development, are consistent with this identification. That is, strong expression of phm is restricted to the prothoracic gland cells of the Drosophila larval ring gland, where it undergoes dramatic changes in expression, and in the adult ovary, but also in the embryonic epidermis. During the last larval-larval transition in Bombyx, a similar expression pattern in the prothoracic gland is observed, but as in Drosophila, slight expression is also present in other tissues, suggesting a possible additional role for the phantom enzyme. PMID- 15350620 TI - Knowledge modeling and acquisition of traditional Chinese herbal drugs and formulae from text. AB - Traditional Chinese medicine has developed over more than 4000 years. A tremendous amount of medical knowledge has been accumulated, among which herbal drugs and formulae are an important portion. This paper presents an ontology for traditional Chinese drugs and formulae, and an ontology-based system for extracting knowledge of drugs and formulae from semi-structured text. The system consists of two components: an executable knowledge extraction language (or EKEL) for specifying knowledge-extracting agents, and a support machine for executing EKEL programs. Experiments show that the system is adequate of extracting knowledge of herbal drugs and formulae from semi-structured text. PMID- 15350621 TI - Ontology development for unified traditional Chinese medical language system. AB - Traditional Chinese medicine (TCM) as a complete knowledge system researches into human health conditions via a different approach compared to orthodox medicine. We are developing a unified traditional Chinese medical language system (UTCMLS) through an ontology approach that will support TCM language knowledge storage, concept-based information retrieval and information integration. UTCMLS is a huge knowledge project, which is a broad collaboration of 16 distributed groups, most of them with no prior experience of formal ontology development. Therefore, the cooperative and comprehensive ontology engineering is crucial. We use Protege 2000 for ontology development of concepts and relationships that represent the domain and that will permit storage of TCM knowledge. This paper focuses on the methodology, design and development of ontology for UTCMLS. PMID- 15350622 TI - Local fuzzy fractal dimension and its application in medical image processing. AB - The local fuzzy fractal dimension (LFFD) is proposed to extract local fractal feature of medical images. The definition of LFFD is an extension of the pixel covering method by incorporating the fuzzy set. Multi-feature edge detection is implemented with the LFFD and the Sobel operator. The LFFD can also serve as a characteristic of motion in medical image sequences. The experimental results show that the LFFD is an important feature of edge areas in medical images and can provide information for segmentation of echocardiogram image sequences. PMID- 15350623 TI - A novel kernelized fuzzy C-means algorithm with application in medical image segmentation. AB - Image segmentation plays a crucial role in many medical imaging applications. In this paper, we present a novel algorithm for fuzzy segmentation of magnetic resonance imaging (MRI) data. The algorithm is realized by modifying the objective function in the conventional fuzzy C-means (FCM) algorithm using a kernel-induced distance metric and a spatial penalty on the membership functions. Firstly, the original Euclidean distance in the FCM is replaced by a kernel induced distance, and thus the corresponding algorithm is derived and called as the kernelized fuzzy C-means (KFCM) algorithm, which is shown to be more robust than FCM. Then a spatial penalty is added to the objective function in KFCM to compensate for the intensity inhomogeneities of MR image and to allow the labeling of a pixel to be influenced by its neighbors in the image. The penalty term acts as a regularizer and has a coefficient ranging from zero to one. Experimental results on both synthetic and real MR images show that the proposed algorithms have better performance when noise and other artifacts are present than the standard algorithms. PMID- 15350624 TI - An efficient and scalable deformable model for virtual reality-based medical applications. AB - Modeling of tissue deformation is of great importance to virtual reality (VR) based medical simulations. Considerable effort has been dedicated to the development of interactively deformable virtual tissues. In this paper, an efficient and scalable deformable model is presented for virtual-reality-based medical applications. It considers deformation as a localized force transmittal process which is governed by algorithms based on breadth-first search (BFS). The computational speed is scalable to facilitate real-time interaction by adjusting the penetration depth. Simulated annealing (SA) algorithms are developed to optimize the model parameters by using the reference data generated with the linear static finite element method (FEM). The mechanical behavior and timing performance of the model have been evaluated. The model has been applied to simulate the typical behavior of living tissues and anisotropic materials. Integration with a haptic device has also been achieved on a generic personal computer (PC) platform. The proposed technique provides a feasible solution for VR-based medical simulations and has the potential for multi-user collaborative work in virtual environment. PMID- 15350625 TI - RET polymorphisms in codons 769 and 836 are not associated with predisposition to medullary thyroid carcinoma. AB - The study was undertaken to verify whether the RET gene polymorphisms are associated with MTC in patients negative for germline mutations. Two hundred five patients with apparent sporadic MTC were subjected to genetic analysis of RET exons 10, 11, 13, 14, 16 and 22 RET germline mutation carriers were identified with 10.7% frequency. The frequency among 26 patients not older than 30 was 27%. In patients excluded for known mutations we analyzed two polymorphic sites: RET codon 769 and 836. As control group, 90 healthy subjects were investigated. In young patients the observed allelic frequencies were 32% for variant L769/CTG and 5% for variant S836/AGT. Although these values were higher than in older MTC patients (22 and 3%, respectively), as well as in the control group (27 and 2%) the difference was insignificant. We conclude that in Polish patients polymorphisms at RET codons 769 and 836 are not associated with medullary thyroid carcinoma. PMID- 15350626 TI - Cyclin D1 A870G polymorphism and amplification in laryngeal squamous cell carcinoma: implications of tumor localization and tobacco exposure. AB - Altered Cyclin D1 activity, due to gene amplification and/or protein overexpression, is related to the development of several human cancers, including head and neck SCC. This study investigated the relationship between CCND1 A870G gene polymorphism and amplification with the development and progression of laryngeal SCC, considering the implications of tumor localization and tobacco exposure. The study population consisted of 66 larynx cancer patients and 110 healthy individuals. CCND1 A/G polymorphism in exon 4 was genotyped by a PCR-RFLP assay. Cyclin D1 gene amplification was evaluated by a Differential-PCR assay and determined by a quantitative densitometric analysis. Our data on gene amplification did not show any correlation with disease stage, histological tumor differentiation, recurrent disease, disease-specific survival or tumor location. However, GG870 genotype was associated with a shorter disease free interval and a reduced overall survival in laryngeal cancer patients. Moreover, this constitutes the first report of a correlation between cyclin D1 A870G polymorphism and increased susceptibility for laryngeal tumor development at the glottic region, which supports the theory of site-specific prevalence of genetic alterations. PMID- 15350627 TI - Multi-target interphase fluorescence in situ hybridization assay increases sensitivity of sputum cytology as a predictor of lung cancer. AB - Survival rates for lung cancer are low because patients have disseminated disease at diagnosis; therefore tests for early diagnosis are highly desirable. This pilot study investigated occurrence of chromosomal aneusomy in sputum from a 33 case-control cohort matched on age, gender, and date of sample collection. Subjects had chronic obstructive pulmonary disease and > or = 30 pack-years of tobacco use, and aneusomy was tested using a multi-target DNA FISH assay (LAVysion, Abbott/Vysis). In specimens collected within 12 months of lung cancer diagnosis, abnormality was more frequent among the 18 cases (41%) than the 17 controls (6%; P = 0.04). Aneusomy had no significant association with cytologic atypia, which might indicate that molecular and morphological changes could be independent markers of tumorigenesis. Combining both tests, abnormality was found in 83% of the cases and 20% of the controls (P = 0.0004) suggesting that FISH may improve the sensitivity of cytologic atypia as a predictor of lung cancer. PMID- 15350628 TI - p53 protein accumulation in colorectal cancer tissue has prognostic value only in left-sided colon tumours. AB - The aim of this paper is to describe the prospective 10-year follow-up study of prognostic value of p53 protein accumulation in cancer cells in relation to the localisation of tumour in colorectum. METHODS: The p53 protein accumulation has been assessed immunohistochemically in tumour tissue samples obtained from 80 selected sporadic colorectal cancer patients. RESULTS: p53 protein accumulation was detected more often in rectum than in colon tumours. Lower survival rates were observed in patients with p53 protein accumulation in cancer cells independent of primary tumour site. Significantly lower cumulative survival rate was detected in the group of patients with left-sided colon cancer. The p53 protein accumulation was an independent prognostic factor in the left-sided colon tumours. CONCLUSIONS: The right-sided colon tumours may be developed in p53 independent manner and p53 protein accumulation in cancer cells has prognostic value only in the left-sided colorectal tumours. PMID- 15350629 TI - Effect of physician recommendation and patient adherence on rates of colorectal cancer testing. AB - This study explored: (1) patient characteristics associated with physician recommendation for colorectal cancer (CRC) screening and patient adherence to recommendation, and (2) the combined effect of recommendation and adherence on CRC testing, broadly defined. Data were from the 1999 MA BRFSS and a call-back survey of 869 BRFSS participants, age 50 and older. Logistic regression was used to identify correlates of recommendation, adherence, and testing. Patient physician factors were positively associated with recommendation, adherence and testing. Inadequate health insurance was negatively associated with recommendation (OR = 0.45, 95% CI = 0.27-0.78) and testing (OR = 0.64, 95% CI = 0.38-1.1). Men were not more likely to be recommended (OR = 1.1, 95% CI = 0.78 1.5), but were more likely to adhere (OR = 1.9, 95% CI = 1.2-2.0) and to be tested (OR = 1.4, 95% CI = 1.0-1.9). There were gender differences in recommendation when considering health and risk factor measures. Research is needed to understand differences in recommendation and adherence. Greater encouragement and follow-through may be needed for groups less likely to adhere. PMID- 15350630 TI - Personal cancer prevention and screening practices among Asian Indian physicians in the United States. AB - This study examines personal cancer prevention and screening practices of Asian Indian physicians. Asian Indians are the third largest group of Asian Pacific Islanders (APIs) residing in the United States. Using a cross-sectional study approach, we investigated cancer prevention and screening practices of 254 randomly selected Asian Indian physicians in the United States. Results showed that prevalence of conventional risk factors were low, e.g., smoking, alcohol consumption. Nutrition-related behaviors indicated 31-48% met the Food Guide Pyramid's recommendations for daily intake of fruits, vegetables, low fat/cholesterol diet, and grains. Certain cancer prevention and screening practices failed to meet age appropriate recommendations of leading health agencies. For men age 50 and over, only 27% ever had a flexible sigmoidoscopy and 25% ever had a fecal occult blood test (FOBT); 84% of women age 40 and over ever had a mammogram and 87% of women ever had a Papanicalou (Pap) smear. With respect to cancer screening behaviors, the respondents were less likely to engage in these preventive screenings (with the exception of mammography) than the general US population and the APIs (except Pap smears). Additionally, our respondents were less likely than other physicians to practice cancer screening behaviors. Results support the need to promote regular cancer prevention and screening among Asian Indian physicians. PMID- 15350631 TI - Rapid method for identification of chemopreventive compounds using multiplex RT PCR for cyclooxygenase mRNA expression. AB - Inhibition of the expression of the cyclooxygenase-2 (COX-2) enzyme has been associated with prevention or reversal of cancer development in several organs. Development of, or screening for, selective cyclooxygenase (COX) inhibitors is an approach for identifying chemopreventive agents. The aim of this project was to develop a rapid semi-quantitative multiplex RT-PCR for analysis of the expression of cyclooxygenase 1 and 2 genes relative to 18S, the housekeeping gene, in a 3 gene multiplex reaction. The optimal conditions for the 3-gene multiplex reaction were determined experimentally. Following RT-PCR, the amplified PCR products are analyzed by capillary electrophoresis-based LabChip and the Agilent 2100 bioanalyzer, which eliminates electrophoresis through ethidium bromide-agarose gels and normalization of band intensities using software programs such as Molecular Analyst. The multiplex RT-PCR assay developed here is a cost effective technique for routine application in laboratories that are screening natural or synthetic compounds for specific inhibitors of cyclooxygenase-2 as potential chemopreventive agents. PMID- 15350632 TI - The activity of latent benzoperimidine esters to inhibit P-glycoprotein and multidrug resistance-associated protein 1 dependent efflux of pirarubicin from several lines of multidrug resistant tumor cells. AB - Multidrug resistance of tumor cells is associated with the presence of membrane proteins responsible for the cytostatics export. Recently, we have synthesized a new family of benzoperimidines causing the futile cycle of MDR pumps. In this study, biological data for benzoperimidine esters are presented for selected cell lines: sensitive (HL-60, GLC4, K562), P-gp resistant (HL-60/VINC, K562/DX), MRP1 resistant (HL-60/DX) and MRP1/LRP resistant (GLC4/DX). Their ability to inhibit the efflux of anthracycline antitumor drug, pirarubicin and to restore its accumulation in MDR cells was studied using a spectrofluorometric method which allows to follow the uptake and efflux of fluorescent molecules by living cells. Benzoperimidine esters had high effectiveness in inhibiting pirarubicin efflux and in restoring its accumulation in resistant cells. In contrast, examined esters were less active in vitro in restoration of pirarubicin cytotoxicity towards resistant cells because an enzymatic cleavage of esters occurs in presence of serum esterases. PMID- 15350633 TI - Are there geographical differences in the frequency of SYT-SSX1 and SYT-SSX2 chimeric transcripts in synovial sarcoma? AB - Synovial sarcoma (SS) is characterized by the t(X;18)(p11.2;q11.2) chromosomal translocation, which results in generating either SYT-SSX1, SYT-SSX2 or, infrequently, SYT-SSX4 fusion gene. The ratio of SYT-SSX1:SYT-SSX2 fusions is close to 2:1 in the majority of studies, and SYT-SSX2 fusion has been only rarely observed in biphasic SS. In the present study, we compared two series of patients with SS, Slovenian (37 cases) and Dutch (14 cases), with respect to clinical, pathological and molecular findings. The two groups did not differ with regard to clinicopathological features. Whereas the frequency of different SYT-SSX fusions in the Dutch group was similar to that reported in the literature, we found an unexpectedly high number of tumors with SYT-SSX2 fusion in the Slovenian group. The ratio of SYT-SSX1:SYT-SSX2 fusion was 7:18 for monophasic and 2:7 for biphasic tumors in the Slovenian group. This distribution differs significantly from that observed in the Dutch group in the present study (P = 0.041) as well as from data reported in the recent large multi-institutional study on 243 patients (P = 0.0001). Our findings indicate possible geographical differences in the frequency of two SYT-SSX fusion transcripts in patients with synovial sarcoma. PMID- 15350634 TI - Geographic pattern of cancers related to tobacco and alcohol in Connecticut: implications for cancer control. AB - The purpose of this study was to examine the geographic distribution of standardized incidence ratios (SIRs) for Connecticut's 169 towns for 18,382 cancers diagnosed in 1995-2000 at sites most strongly associated with tobacco and/or alcohol (i.e, lung, oral cavity-pharynx, and esophagus), with consideration of census-derived indicators of socioeconomic status (SES) at the town level. For males, the state's four largest towns, all in the highest poverty rate quartile, had statistically significantly elevated SIRs for both lung cancer and oral cavity-pharynx cancers, and also had elevated SIRs for esophageal cancer. Two of these four towns also had statistically significantly elevated SIRs for oral cavity-pharynx cancer for females. SIRs for both males and females were lowest for the lowest poverty quartile and highest for the highest poverty quartile, for each cancer-site group. Among 15,271 patients diagnosed with their first cancer at any of the selected sites in 1995-2000, risk of diagnosis of a second primary cancer at any of these sites (139 patients) was highest in the highest poverty-rate quartile. These surveillance methods should be useful for targeting cancer control efforts aimed at prevention or cessation of tobacco and/or heavy alcohol use, and early detection or chemoprevention of these cancers, including second primary cancers. PMID- 15350635 TI - Targeting of GLUR4-containing AMPA receptors to synaptic sites during in vitro classical conditioning. AB - The synaptic delivery of GluR4-containing AMPA receptors during in vitro classical conditioning of a neural correlate of an eyeblink response was examined by fluorescence imaging of punctate staining for glutamate receptor subunits and the presynaptic marker synaptophysin. There was a significant increase in GluR4 containing AMPA receptors to synaptic sites after conditioning as determined by colocalization of GluR4 subunit puncta with synaptophysin. Moreover, the trafficking of these receptor subunits requires NMDA receptor activation as it was blocked by D,L-2-amino-5-phosphonovaleric acid (AP-5). In contrast, colocalization of NR1 subunits with synaptophysin was stable regardless of whether the preparations had undergone conditioning or had been treated by AP-5. The enhanced colocalization of GluR4 and synaptophysin was accompanied by an increase in both the total number and size of puncta for both proteins, suggesting greater synthesis and aggregation during conditioning. Western blot analysis confirmed upregulation of synaptophysin and GluR4 following conditioning. These data support the hypothesis that GluR4-containing AMPA receptors are delivered to synaptic sites during conditioning. Further, they suggest coordinate presynaptic and postsynaptic modifications during in vitro classical conditioning. PMID- 15350636 TI - Role of glutamate receptors in nucleus accumbens core and shell in spatial behaviour of rats. AB - The nucleus accumbens (NAC) is considered to be an important neural interface between corticolimbic and motor systems of the brain. Several studies have shown that the NAC is not only involved in motivation and reward-related processes but also in spatial behavior. We here investigated the involvement of different glutamate receptor subclasses within NAC core and shell subregions on behavior in a radial-maze. Rats were first trained in a four-arm-baited eight-arm radial maze task for baseline performance. Thereafter, the effects of microinjection of the nonselective glutamate receptor antagonist kynurenic acid (4.5 microg), the NMDA receptor antagonist 2-amino-5-phosphonopentanoic acid (1 microg) and the non-NMDA receptor antagonist 6,7-dinitroquinoxaline-2,3-dione (0.75 microg) in NAC core and shell were tested on reference memory errors (RME) and working memory errors (WME). Moreover, the choice pattern of entries and duration of arm-entries were evaluated. Microinjection of all drugs increased RME. Additionally, non-NMDA receptor blockade in NAC shell but not core increased WME. After microinjection of all drugs into NAC core and shell rats preferentially choose the arms next to the previously visited arm. This work shows that glutamate receptors in both NAC subregions are important for spatial behavior. The deficits seen after glutamate receptor blockade may not be working- or reference memory-related but caused by a switch from a memory-dependent allocentric strategy to an egocentric response strategy. PMID- 15350637 TI - Metabolic cholesterol depletion hinders cell-surface trafficking of the nicotinic acetylcholine receptor. AB - The effects of metabolic inhibition of cholesterol biosynthesis on the trafficking of the nicotinic acetylcholine receptor (AChR) to the cell membrane were studied in living CHO-K1/A5, a Chinese hamster ovary clonal line that heterologously expresses adult alpha2betadeltaepsilon mouse AChR. To this end, we submitted CHO-K1/A5 cells to long-term cholesterol deprivation, elicited by Mevinolin, a potent inhibitor of 3-hydroxy-3-methyl-glutaryl-CoA reductase and applied a combination of biochemical, pharmacological and fluorescence microscopy techniques to follow the fate of the AChR. When CHO-K1/A5 cells were grown for 48 h in lipid-deficient medium supplemented with 0.5 microM Mevinolin, total cholesterol was significantly reduced (40%). Concomitantly, the maximum number of binding sites (Bmax) of the cell-surface AChR for the competitive antagonist alpha-bungarotoxin was reduced from 647+/-30 to 352+/-34 fmol/mg protein, i.e. by 46%. The apparent dissociation constant (Kdapp) for alpha-bungarotoxin of the AChRs remaining at the cell surface was not modified by cholesterol depletion. Similarly, the half-concentration inhibiting the specific binding of the radioligand (IC50) for another competitive antagonist, d-tubocurarine, did not differ from that in control cells. The decrease in cell-surface AChR was paralleled by an increase in intracellular AChR levels, which rose from 44+/-2.1% in control cells to 74+/-3.3% in Mevinolin-treated cells. When analyzed by wide field fluorescence microscopy, the fluorescence signal arising from alpha bungarotoxin labeled cell-surface AChRs was reduced by approximately 70% in Mevinolin-treated cells. The distribution of intracellular AChR also changed: Alexa594-alpha-bungarotoxin-labeled AChR exhibited a highly compartmentalized pattern, concentrating at the perinuclear and Golgi-like regions. Temperature arrest of protein trafficking magnified this effect, emphasizing the Golgi localization of the AChR. Colocalization studies using the transiently expressed fluorescent trans-Golgi/trans-Golgi network marker pEYFP/human beta1,4 galactosyltransferase and the trans-Golgi network marker syntaxin 6 provided additional support for the Golgi localization of intracellular AChRs. The low AChR cell-surface expression and the increase in intracellular AChR pools in cholesterol-depleted cells raise the possibility that cholesterol participates in the trafficking of the receptor protein to the plasmalemma and its stability at this surface location. PMID- 15350638 TI - Remodelling of synaptic morphology but unchanged synaptic density during late phase long-term potentiation (LTP): a serial section electron micrograph study in the dentate gyrus in the anaesthetised rat. AB - In anaesthetised rats, long-term potentiation (LTP) was induced unilaterally in the dentate gyrus by tetanic stimulation of the perforant path. Animals were killed 6 h after LTP induction and dendritic spines and synapses in tetanised and untetanised (contralateral) hippocampal tissue from the middle molecular layer (MML) were examined in the electron microscope using stereological analysis. Three-dimensional reconstructions were also used for the first time in LTP studies in vivo, with up to 130 ultrathin serial sections analysed per MML dendritic segment. A volume sampling procedure revealed no significant changes in hippocampal volume after LTP and an unbiased counting method demonstrated no significant changes in synapse density in potentiated compared with control tissue. In the potentiated hemisphere, there were changes in the proportion of different spine types and their synaptic contacts. We found an increase in the percentage of synapses on thin dendritic spines, a decrease in synapses on both stubby spines and dendritic shafts, but no change in the proportion of synapses on mushroom spines. Analysis of three-dimensional reconstructions of thin and mushroom spines following LTP induction revealed a significant increase in their volume and area. We also found an increase in volume and area of unperforated (macular) and perforated (segmented) postsynaptic densities. Our data demonstrate that whilst there is no change in synapse density 6 h after the induction of LTP in vivo, there is a considerable restructuring of pre-existing synapses, with shaft and stubby spines transforming to thin dendritic spines, and mushroom spines changing only in shape and volume. PMID- 15350639 TI - Circadian tracking of nicotinamide cofactor levels in an immortalized suprachiasmatic nucleus cell line. AB - Nicotinamide adenine dinucleotides can exhibit a daily rhythm in plants and regulate the activity of mammalian clock-like transcription factors in vitro. Because one such redox-sensitive transcription factor is present in the master circadian clock of the brain (the suprachiasmatic nuclei, SCN) and the SCN exhibits a characteristic daily rhythm in glucose usage, nicotinamide cofactors might be expected to influence, exhibit, and/or reflect biological rhythms in SCN cells. Therefore, cofactors were extracted from a model SCN cell line at 3 h intervals over 1-2 day periods and samples were analyzed by capillary electrophoresis with multiphoton excitation of fluorescence. Natively fluorescent reduced cofactors (nicotinamide adenine dinucleotide, NADH, and its phosphorylated form, NADPH) were assayed directly, and nonfluorescent oxidized cofactors (nicotinamide adenine dinucleotide, NAD, and its phosphorylated form, NADP) were enzymatically reduced to their fluorescent counterparts before analysis. In the first day after a synchronizing pulse of fetal bovine serum, a dramatic upregulation in cellular NADH content was observed, consistent with a response to serum insulin; this was accompanied by a smaller decrease in NADPH redox state, which may indicate scavenging of reactive oxygen species generated by increased cellular metabolism. However, when cells were investigated after these early phenomena had recovered or stabilized, no circadian NAD(P)(H) rhythms were observed. During these studies, the NADH/NAD(H) concentration ratio in SCN2.2 cells (0.13+/-0.03) was not high enough to activate clock-like transcription factors. Although the NADPH/NADP(H) concentration ratio was more appropriate (0.8+/-0.1), the intracellular NADPH concentration was < or = 0.7 mM, far too low for half-maximal DNA binding of clock-like transcription factors in vitro. Moreover, these concentration and ratio values represent cellular averages, and free cofactors should be much lower in the cell nucleus. Our data show that SCN2.2 cells maintain nearly constant circadian NAD(P)(H) levels, and that the previously reported in vitro relationship between clock-like transcription factors and NAD(P)(H) does not appear to be biologically relevant. PMID- 15350640 TI - P2X7-like receptor subunits enhance excitatory synaptic transmission at central synapses by presynaptic mechanisms. AB - Recent studies demonstrate that P2X7 receptor subunits (P2X7RS) are present at central and peripheral synapses and suggest that P2X7RS can regulate transmitter release. In brainstem slices from 15 to 26 day old pentobarbitone-anesthetized mice, we examined the effect of P2X7RS activation on excitatory postsynaptic currents (EPSCs) recorded from hypoglossal motoneurons using whole-cell patch clamp techniques. After blockade of most P2X receptors with suramin (which is inactive at P2X7RS) and of adenosine receptors with 8-phenyltheophylline (8PT), bath application of the P2X receptor agonist 3'-0-(4-benzoyl)ATP (BzATP) elicited a 40.5+/-16.0% (mean+/-S.E.M., n = 8, P = 0.039) increase in evoked EPSC amplitude and significantly reduced paired pulse facilitation of evoked EPSCs. This response to BzATP (with suramin and 8PT present) was completely blocked by prior application of Brilliant Blue G (200 nM or 2 microM), a P2X7RS antagonist. In contrast, BzATP application with suramin and 8PT present did not alter miniature EPSC frequency or amplitude when action potentials were blocked with tetrodotoxin. These electrophysiological results suggest that P2X7RS activation increases central excitatory transmitter release via presynaptic mechanisms, confirming previous indirect measures of enhanced transmitter release. We suggest that possible presynaptic mechanisms underlying enhancement of evoked transmitter release by P2X7RS activation are modulation of action potential width or an increase in presynaptic terminal excitability, due to subthreshold membrane depolarization which increases the number of terminals releasing transmitter in response to stimulation. PMID- 15350641 TI - Proteasomal inhibition induced by manganese ethylene-bis-dithiocarbamate: relevance to Parkinson's disease. AB - Maneb, a widely used fungicide, has been associated with Parkinsonism in humans. In experimental models, maneb and its major active element, manganese ethylene bis-dithiocarbamate (Mn-EBDC) cause selective nigrostriatal neurodegeneration in mice and in rats, respectively. To investigate the mechanisms underlying this neurodegeneration, we studied the effects of Mn-EBDC on proteasomal function, which is decreased in patients with Parkinson's disease (PD), in a dopaminergic neuronal cell line (MES 23.5 or MES). The results demonstrated that exposure of MES cells to 6 microM Mn-EBDC for 7 days produced not only significant neurotoxicity but also inhibition of proteasomal chymotrypsin-like and postglutamyl peptidase activities. Proteasomal dysfunction was accompanied by formation of cytoplasmic inclusions that were positive for alpha-synuclein immunostaining and significantly increased sodium dodecyl sulfate-insoluble alpha synuclein aggregation seen by Western blot analysis. In addition, there was a significant increase in oxidative stress, evidenced by elevated total protein carbonyl content, in cells treated with Mn-EBDC. Manipulation of intracellular reduced glutathione levels with N-acetyl-L-cysteine or L-buthionine sulfoximine pretreatment to modulate Mn-EBDC-mediated oxidative stress altered Mn-EBDC mediated neurotoxicity, proteasomal dysfunction, and alpha-synuclein aggregation in these cells. These data suggest that neurotoxicity-induced by Mn-EBDC is at least partially attributable to Mn-EBDC-mediated proteasomal inhibition, and that the proteasome may be an important target by which environmental exposure modifies the risk for developing PD in vulnerable populations. PMID- 15350642 TI - Acetyl-L-carnitine induces a sustained potentiation of the afterhyperpolarization. AB - Acetyl-L-carnitine is known to improve many aspects of the neural activity even if its exact role in neurotransmission is still unknown. This study investigates the effects of acetyl-L-carnitine in T segmental sensory neurons of the leech Hirudo medicinalis. These neurons are involved in some forms of neural plasticity associated with learning processes. Their physiological firing is accompanied by a large afterhyperpolarization that is mainly due to the Na+/K+ ATPase activity and partially to a Ca2+ -dependent K+ current. A clear-cut hyperpolarization and a significant increase of the afterhyperpolarization have been recorded in T neurons of leeches injected with 2 mM acetyl-L-carnitine some days before. Acute treatments of 50 microM acetyl-L-carnitine induced similar effects in T cells of naive animals. In the presence of apamin, a pharmacological blocker of Ca2+ dependent K+ channel, acetyl-L-carnitine still enhanced the residual afterhyperpolarization, suggesting an effect of the drug on the Na+/K+ATPase. Acetyl-L-carnitine also increased the hyperpolarization induced by intracellular injection of Na+ ions. Therefore, acetyl-L-carnitine seems to be able to exert a positive sustained effect on the Na+/K+ ATPase activity in leech T sensory neurons. Moreover, in these cells, widely arborized, the afterhyperpolarization seems to play an important role in determining the action potential transmission at neuritic bifurcations. A computational model of a T cell has been previously developed considering detailed data for geometry and the modulation of the pump current. Herein, we showed that to a larger afterhyperpolarization, due to the acetyl-L-carnitine-induced effects, corresponds a decrement in the number of action potentials reaching synaptic terminals. PMID- 15350643 TI - Injury-induced alterations in N-methyl-D-aspartate receptor subunit composition contribute to prolonged 45calcium accumulation following lateral fluid percussion. AB - Cells that survive traumatic brain injury are exposed to changes in their neurochemical environment. One of these changes is a prolonged (48 h) uptake of calcium which, by itself, is not lethal. The N-methyl-D-aspartate receptor (NMDAR) is responsible for the acute membrane flux of calcium following trauma; however, it is unclear if it is involved in a flux lasting 2 days. We proposed that traumatic brain injury induced a molecular change in the NMDAR by modifying the concentrations of its corresponding subunits (NR1 and NR2). Changing these subunits could result in a receptor being more sensitive to glutamate and prolong its opening, thereby exposing cells to a sustained flux of calcium. To test this hypothesis, adult rats were subjected to a lateral fluid percussion brain injury and the NR1, NR2A and NR2B subunits measured within different regions. Although little change was seen in NR1, both NR2 subunits decreased nearly 50% compared with controls, particularly within the ipsilateral cerebral cortex. This decrease was sustained for 4 days with levels returning to control values by 2 weeks. However, this decrease was not the same for both subunits, resulting in a decrease (over 30%) in the NR2A:NR2B ratio indicating that the NMDAR had temporarily become more sensitive to glutamate and would remain open longer once activated. Combining these regional and temporal findings with 45calcium autoradiographic studies revealed that the degree of change in the subunit ratio corresponded to the extent of calcium accumulation. Finally, utilizing a combination of NMDAR and NR2B-specific antagonists it was determined that as much at 85% of the long term NMDAR-mediated calcium flux occurs through receptors whose subunits favor the NR2B subunit. These data indicate that TBI induces molecular changes within the NMDAR, contributing to the cells' post-injury vulnerability to glutamatergic stimulation. PMID- 15350644 TI - Zac1 is up-regulated in neural cells of the limbic system of mouse brain following seizures that provoke strong cell activation. AB - Zac1, a new zinc-finger protein that regulates both apoptosis and cell cycle arrest, is abundantly expressed in many proliferative/differentiation areas during brain development. In the present work, we studied Zac1 gene expression and protein in experimental seizure models following i.p. injection of pentylenetetrazole (PTZ) or kainic acid (KA). Following KA treatment, an early and intense up-regulation of Zac1 is detected in the limbic areas, such as the hippocampus, cortex and amygdaloid and hypothalamic nuclei. Pre-treatment with MK 801, an antagonist of the NMDA receptors, fully blocks the effect of KA in the hippocampus, whereas it only attenuates KA-induced Zac1 up-regulation in the other areas of the limbic system. A reduced induction is obtained with PTZ treated animals, specifically in the entorhinal and piriform cortices as well as in amygdaloid and hypothalamic nuclei. Thus, Zac1 is highly induced in the seizure models that generate strong neuronal stimulation and/or extensive cell damage (cell death), reinforcing its putative role in the control of the cell cycle and/or apoptosis. Moreover, strong induction is observed in the granular cells of the dentate gyrus (which are resistant to neurodegeneration) and in some glial cells of the dentate gyrus and subventricular zone, suggesting that Zac1 may be implicated in the mechanisms of neural plasticity following injury. PMID- 15350645 TI - Hypoxic increase in nitric oxide generation of rat sensory neurons requires activation of mitochondrial complex II and voltage-gated calcium channels. AB - Recently, we have demonstrated that sensory neurons of rat lumbar dorsal root ganglia (DRG) respond to hypoxia with an activation of endothelial nitric oxide (NO) synthase (eNOS) resulting in enhanced NO production associated with mitochondria which contributes to resistance against hypoxia. Extracellular calcium is essential to this effect. In the present study on rat DRG slices, we set out to determine what types of calcium channels operate under hypoxia, and which upstream events contribute to their activation, thereby focusing upon mitochondrial complex II. Both the metallic ions Cd2+ and Ni2+, known to inhibit voltage-gated calcium channels and T-type channels, respectively, and verapamil and nifedipine, typical blocker of L-type calcium channels completely prevented the hypoxic neuronal NO generation. Inhibition of complex II by thenoyltrifluoroacetone at the ubiquinon binding site or by 3-nitropropionic acid at the substrate binding site largely diminished hypoxic-induced NO production while having an opposite effect under normoxia. An additional blockade of voltage gated calcium channels entirely abolished the hypoxic response. The complex II inhibitor malonate inhibited both normoxic and hypoxic NO generation. These data show that complex II activity is required for increased hypoxic NO production. Since succinate dehydrogenase activity of complex II decreased at hypoxia, as measured by histochemistry and densitometry, we propose a hypoxia-induced functional switch of complex II from succinate dehydrogenase to fumarate reductase, which subsequently leads to activation of voltage-gated calcium channels resulting in increased NO production by eNOS. PMID- 15350646 TI - N-methyl-D-aspartate receptors enhance mechanical responses and voltage-dependent Ca2+ channels in rat dorsal root ganglia neurons through protein kinase C. AB - N-methyl-D-aspartate (NMDA)receptors (NMDARs) located on peripheral terminals of primary afferents are involved in the transduction of noxious mechanical stimuli. Exploiting the fact that both NMDARs and stretch-activated channels are retained in short-term culture and expressed on the soma of dorsal root ganglia (DRG) neurons, we examined the effect of NMDA on mechanically mediated changes in intracellular calcium concentration ([Ca2+]i). Our aims were to determine whether NMDARs modulate the mechanosensitivity of DRG neurons. Primary cultures of adult rat lumbosacral DRG cells were cultured for 1-3 days. [Ca2+]i responses were determined by Fura-2 ratio fluorescence. Somas were mechanically stimulated with fire-polished glass pipettes that depressed the cell membrane for 0.5 s. Voltage activated inward Ca2+ currents were measured by the whole cell patch clamp. Stimulation of neurons with 100 microM NMDA in the presence, but not the absence, of co-agonist (10 microM D-serine) caused transient [Ca2+]i responses (101+/-9 nM) and potentiated [Ca2+]i peak responses to subsequent mechanical stimulation more than two-fold (P < 0.001). NMDA-mediated potentiation of mechanically induced [Ca2+]i responses was inhibited by the selective protein kinase C (PKC) inhibitor GF109203X (GFX; 10 microM), which had no independent effects on NMDA- or mechanically induced responses. Short-term treatment with the PKC activator phorbol dibutyrate (1 microM PDBu for 1-2 min) also potentiated mechanically induced [Ca2+]i responses nearly two-fold (P < 0.001), while longer exposure (>10 min) inhibited the [Ca2+]i transients by 44% (P < 0.001). Both effects of PDBu were prevented by prior treatment with GFX. Inhibition of voltage-dependent Ca2+ channels with 25 microM La3+ had no effect on mechanically induced [Ca2+]i transients prior to NMDA, but prevented enhancement of the transients by NMDA and PDBu. NMDA pretreatment transiently enhanced nifedipine-sensitive, voltage activated Ca2+ currents by a process that was sensitive to GFX. In conclusion, activation of NMDARs on cultured DRG neurons sensitize voltage-dependent L-type Ca2+ channels which contribute to mechanically induced [Ca2+]i transients through a PKC-mediated process. PMID- 15350647 TI - The superoxide dismutase1 (SOD1) G93A mutation does not promote neuronal injury after focal brain ischemia and optic nerve transection in mice. AB - The superoxide dismutase 1 (SOD1)G93A mouse was recently established as transgenic model of amyotrophic lateral sclerosis. We were interested to know whether the SOD1 G93A mutation promotes neuronal injury after intraluminal middle cerebral artery thread occlusion and/or retinal ganglion cell (RGC) axotomy in mice, which are highly reproducible and clinically relevant in vivo models of acute and subacute neuronal degeneration, respectively. In our experiments, G93A mutant SOD1 neither influenced ischemic injury after 90 or 30 min of focal ischemia, nor had an impact on the severity of RGC degeneration after optic nerve transection, when human SOD1 G93A mutant mice were compared to human wild-type SOD1 mice. Our data indicate that the clinically relevant SOD1 G93A mutation, which leads to amyotrophic lateral sclerosis in humans and mice, does not necessarily worsen neuronal degeneration in other pathologies. Thus, the G93A mutation may be counterbalanced in non-motor neurons of young animals, and region specific and age-related factors may be necessary so that neurodegeneration is re enforced. PMID- 15350648 TI - Effects of neonatal excitotoxic lesions of the entorhinal cortex on cognitive functions in the adult rat. AB - The entorhinal cortex (EC) is involved in a variety of cognitive functions by virtue of its neuronal input from the neocortex and projection to the hippocampal formation and the limbic-striatal system. Neonatal lesions are increasingly considered useful models for disconnection syndromes such as schizophrenia. Therefore, we investigated the effects of neonatal EC lesions on adult rat behavior. Neonatal (postnatal day 7) lesions were inflicted by bilateral injections of ibotenate into the EC. Sham-lesioned (vehicle injection) and naive (unoperated) rats served as controls. Locomotor activity was measured in prepubertal and young adult rats. Adult rats were then tested for spatial learning in an eight-arm radial maze (reinforced delayed alternation) and for motivation (progressive ratio schedule of operant behavior). Finally, prepulse inhibition (PPI) of the acoustic startle reflex and locomotor activity were investigated with and without apomorphine (APO) challenge. Brain tissue damage was assessed using Nissl-staining. The total volume of the adult rat EC was reduced after neonatal ibotenate-injection. Neonatal EC-lesions increased perseveration only in a delayed task in the radial maze and induced a leftward shift of breakpoints in operant responding. Lesions did not alter baseline locomotor activity, but enhanced the locomotor stimulating effect of APO. PPI was not affected by neonatal lesions of the EC with and without APO challenge. Neonatal lesions of the EC impaired the ability to hold information during delays and reduced motivation during operant behavior which reflects a state of anhedonia. Thus, they may serve as an animal model for certain aspects of schizophrenia. PMID- 15350649 TI - Differential gene expression after complete spinal cord transection in adult rats: an analysis focused on a subchronic post-injury stage. AB - In an attempt to characterize changes in transcription after a sub-chronic spinal cord injury (SCI), we investigated gene expression profiles using cDNA microarray. Among 7523 genes and expressed sequence tags (ESTs) examined, 444 transcripts, including 218 genes and 226 ESTs, were identified to be either up regulated (373 of 444) or down-regulated (71 of 444) greater than 2.0-fold in the spinal cord at 14 days after a complete spinal transection at the 11th thoracic level in adult rats. Based on their potential function, these differentially expressed genes were categorized into seven classes which include cell division related protein, channels and receptors, cytoskeletal elements, extracellular matrix proteins, metalloproteinases and inhibitors, growth-associated molecules, metabolism, intracellular transducers and transcription factors, as well as others. Strong expressional changes were found in all classes revealing the complexity and diversity of gene expression profiles following SCI. We verified array results with RT-PCR for eight genes, Northern blotting for nine genes, and in situ hybridization for one gene and immunohistochemistry for four genes. These analyses confirmed, to a large extent, that the array results have accurately reflected the molecular changes occurring at 14 days post-SCI. Importantly, the current study has identified a number of genes, including annexins, heparin binding growth-associated protein (HB-GAM), P9ka (S100A4), matrix metalloproteinases, and lysozyme, that may shed new light on SCI-related inflammation, neuroprotection, neurite-outgrowth, synaptogenesis, and astrogliosis. In conclusion, the identification of molecular changes using the large-scale microarray analysis may lead to a better understanding of underlying mechanisms, thus, the development of new repair strategies for SCI. PMID- 15350650 TI - Prostaglandin E2 in the medial preoptic area produces hyperalgesia and activates pain-modulating circuitry in the rostral ventromedial medulla. AB - Prostaglandin E2 (PGE2) produced in the medial preoptic region (MPO) in response to immune signals is generally accepted to play a major role in triggering the illness response, a complex of physiological and behavioral changes induced by infection or injury. Hyperalgesia is now thought to be an important component of the illness response, yet the specific mechanisms through which the MPO acts to facilitate nociception have not been established. However, the MPO does project to the rostral ventromedial medulla (RVM), a region with a well-documented role in pain modulation, both directly and indirectly via the periaqueductal gray. To test whether PGE2 in the MPO produces thermal hyperalgesia by recruiting nociceptive modulating neurons in the RVM, we recorded the effects of focal application of PGE2 in the MPO on paw withdrawal latency and activity of identified nociceptive modulating neurons in the RVM of lightly anesthetized rats. Microinjection of a sub-pyrogenic dose of PGE2 (50 fg in 200 nl) into the MPO produced thermal hyperalgesia, as measured by a significant decrease in paw withdrawal latency. In animals displaying behavioral hyperalgesia, the PGE2 microinjection activated on-cells, RVM neurons thought to facilitate nociception, and suppressed the firing of off-cells, RVM neurons believed to have an inhibitory effect on nociception. A large body of evidence has implicated prostaglandins in the MPO in generation of the illness response, especially fever. The present study indicates that the MPO also contributes to the hyperalgesic component of the illness response, most likely by recruiting the nociceptive modulating circuitry of the RVM. PMID- 15350651 TI - Efferent connections of "posterodorsal" auditory area in the rat cortex: implications for auditory spatial processing. AB - We examined efferent connections of the cortical auditory field that receives thalamic afferents specifically from the suprageniculate nucleus (SG) and the dorsal division (MGD) of the medial geniculate body (MG) in the rat [Neuroscience 117 (2003) 1003]. The examined cortical region was adjacent to the caudodorsal border (4.8-7.0 mm posterior to bregma) of the primary auditory area (area Te1) and exhibited relatively late auditory response and high best frequency, compared with the caudal end of area Te1. On the basis of the location and auditory response property, the cortical region is considered identical to "posterodorsal" auditory area (PD). Injections of biocytin in PD revealed characteristic projections, which terminated in cortical areas and subcortical structures that play pivotal roles in directed attention and space processing. The most noticeable cortical terminal field appeared as dense plexuses of axons in area Oc2M, the posterior parietal cortex. Small terminal fields were scattered in area frontal cortex, area 2 that comprises the frontal eye field. The subcortical terminal fields were observed in the pontine nucleus, the nucleus of the brachium inferior colliculus, and the intermediate and deep layers of the superior colliculus. Corticostriatal projections targeted two discrete regions of the caudate putamen: the top of the middle part and the caudal end. It is noteworthy that the inferior colliculus and amygdala virtually received no projection. Corticothalamic projections terminated in the MGD, the SG, the ventral zone of the ventral division of the MG, the ventral margin of the lateral posterior nucleus (LP), and the caudodorsal part of the posterior thalamic nuclear group (Po). Large terminals were found in the MGD, SG, LP and Po besides small terminals, the major component of labeling. The results suggest that PD is an auditory area that plays an important role in spatial processing linked to directed attention and motor function. The results extend to the rat findings from nonhuman primates suggesting the existence of a posterodorsal processing stream for auditory spatial perception. PMID- 15350652 TI - Differential roles of neuronal and endothelial nitric oxide synthases during carrageenan-induced inflammatory hyperalgesia. AB - The present study investigated the role of neuronal nitric oxide synthase (nNOS) in carrageenan-induced inflammatory pain by combining genomic and pharmacological strategies. Intrathecal injection of the nNOS inhibitor 7-nitroindazole dose dependently inhibited carrageenan-induced thermal hyperalgesia in both early and late phases in wild-type mice. However in nNOS knockout mice, carrageenan-induced thermal hyperalgesia remained intact in the early phase but was reduced in the late phase. Spinal Ca2+ -dependent nitric oxide synthase (NOS) activity in nNOS knockout mice was significantly lower than that in wild-type mice. Following carrageenan injection, although the spinal Ca2+ -dependent NOS activity in both wild-type and knockout mice increased, the enzyme activity in nNOS knockout mice reached a level similar to that in wild-type mice. On the other hand, no significant difference in spinal Ca2+ -independent NOS activity was noted between wild-type and nNOS knockout mice before and after carrageenan injection. Furthermore, intrathecal administration of the endothelial NOS (eNOS) inhibitor L N5-(1-iminoethyl)-ornithinein nNOS knockout mice inhibited the thermal hyperalgesia in both early and late phases, though this inhibitor had no effect in wild-type mice. Meanwhile, Western blot showed that eNOS expression in the spinal cord of nNOS knockout mice was up-regulated compared with wild-type mice; immunohistochemical staining showed that the spinal eNOS was mainly distributed in superficial laminae of the dorsal horn. Finally, double staining with confocal analysis showed that the enhanced spinal eNOS was expressed in astrocytes, but not in neurons. Our current results indicate that nNOS plays different roles in the two phases of carrageenan-induced inflammatory pain. In this model, enhanced spinal eNOS appears to compensate for the role of nNOS in nNOS knockout mice. PMID- 15350653 TI - Patterns of seizures, hippocampal injury and neurogenesis in three models of status epilepticus in galanin receptor type 1 (GalR1) knockout mice. AB - The neuropeptide galanin exhibits anticonvulsant effects in experimental epilepsy. Two galanin receptor subtypes, GalR1 and GalR2, are present in the brain. We examined the role of GalR1 in seizures by studying the susceptibility of GalR1 knockout (KO) mice to status epilepticus (SE) and accompanying neuronal injury. SE was induced in GalR1 KO and wild type (WT) mice by Li-pilocarpine, 60 min electrical perforant path stimulation (PPS), or systemic kainic acid (KA). Seizures were analyzed using Harmonie software. Cell injury was examined by FluoroJade B- and terminal deoxynucleotidyl transferase-mediated uridine triphosphate nick end labeling; neurogenesis was studied using bromodeoxyuridine labeling. Compared with WT littermates, GalR1 KO showed more severe seizures, more profound injury to the CA1 pyramidal cell layer, as well as injury to hilar interneurons and dentate granule cells upon Li-pilocarpine administration. PPS led to more severe seizures in KO, as compared with WT mice. No difference in the extent of neuronal degeneration was observed between the mice of two genotypes in CA1 pyramidal cell layer; however, in contrast to WT, GalR1 KO developed mild injury to hilar interneurons on the side of PPS. KA-induced seizures did not differ between GalR1 KO and WT animals, and led to no injury to the hippocampus in either of experimental group. No differences were found between KO and WT mice in both basal and seizure-induced neuronal progenitor proliferation in all seizure types. Li-pilocarpine led to more extensive glia proliferation in GalR1 KO than in WT, and in both mouse types in two other SE models. In conclusion, GalR1 mediate galanin protection from seizures and seizure-induced hippocampal injury in Li-pilocarpine and PPS models of limbic SE, but not under conditions of KA-induced seizures. The results justify the development and use of GalR1 agonists in the treatment of certain forms of epilepsy. PMID- 15350654 TI - Control of firing by small (S)-alpha-amino-3-hydroxy-5-methyl-isoxazolepropionic acid-like inputs in hypothalamic gonadotropin releasing-hormone (GnRH) neurons. AB - Episodic release of gonadotropin releasing hormone (GnRH) is obligatory for mammalian reproduction. The contribution of synaptic input to intermittent firing of GnRH neurons is unclear. GnRH neurons have very few synapses and most post synaptic currents are small. Therefore, the impact of synaptic currents on firing in GnRH neurons was directly examined using simulated (S)-alpha-amino-3-hydroxy-5 methyl-isoxazolepropionic acid (AMPA)-like inputs applied with the method of dynamic current clamping. Tightly synchronized inputs and 50 ms bursts of excitatory input resulted in action potentials that were coincident with the stimulus. Neither input pattern resulted in sustained firing. When ongoing patterns of simulated inputs were applied over a range of parameters, action potentials were associated with clusters of AMPA-like inputs of 250 pS (approximately 15 pA amplitudes), while single inputs of 500 pS (approximately 30 pA amplitudes) resulted in action potentials. Ongoing inputs of 500 pS drove firing at 4-9 Hz. These findings provide evidence that small, simulated glutamatergic inputs can control firing in GnRH neurons and suggest that despite the small amplitudes, endogenous synaptic input mediated by glutamate may contribute to firing in GnRH neurons. PMID- 15350655 TI - Interleukin-18 null mice show diminished microglial activation and reduced dopaminergic neuron loss following acute 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine treatment. AB - Recent reports have revealed an involvement of microglial cells in dopaminergic neurodegeneration. In the present study, we tested the hypothesis that interleukin-18 (IL-18) plays a role in the microglial activation. The present study investigated microglial activation and dopaminergic neurodegeneration in substantia nigra pars compacta (SNpc) following 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) treatment in wild type (WT) and IL-18 knockout (KO) mice. The number of dopaminergic neuron loss in WT mice was significantly decreased 7 days after MPTP treatment compared with IL-18 KO mice. In WT mice microglial activation occurred in the SN at 1 day after MPTP treatment, progressively increased within the SNpc until 7 days post MPTP, and subsided by 14 days. In contrast, in IL-18 KO mice microglial activation occurred in the SN at 1 day post-MPTP, and decreased by 7 days, earlier than in WT mice. The lesser microglial activation and dopaminergic neurodegeneration in the SNpc following MPTP treatment in WT indicates the possibility that IL-18 may participate in microglial activation and dopaminergic neurodegeneration. PMID- 15350656 TI - Prevalence of Neospora caninum antibodies and factors associated with their presence in dairy cattle of the north of Parana state, Brazil. AB - To determine the prevalence of Neospora caninum antibodies and associated factors, blood sera from 623 female dairy cattle from 23 farms in the north of the state of Parana, Brazil, were analyzed by means of the indirect immunofluorescent-antibody test (IFAT > or = 25). Serum samples from 134 dogs living on the same farms also were tested for N. caninum antibodies (IFAT > or = 50), and the presence of dogs was associated with the prevalence of N. caninum antibodies in cattle. The overall seroprevalence in cattle was 14.3%, mainly in animals over 24 months of age. Seroprevalence in Holsteins (15.1% of 558) was greater than in mixed-breed cattle (7.7% of 65). Age (> or =24 months) of cattle, feeding silage and/or concentrate produced on the farm were associated; antibodies were found in 21.6% of dogs; and the presence of dogs was associated with the prevalence of N. caninum antibodies in cattle. PMID- 15350657 TI - Efficacy of atovaquone against Babesia gibsoni in vivo and in vitro. AB - The therapeutic efficacy of atovaquone against Babesia gibsoni was examined in three dogs experimentally infected with B. gibsoni isolated from naturally infected dogs in Aomori Prefecture, Japan. Once parasitemia reached 10%, atovaquone was administered orally (30 mg/kg twice daily for 7 days). Within 2 days of atovaquone treatment, the parasite disappeared from blood smears without any clinical side effects. Anemia and thrombocytopenia were significantly improved in all the dogs. However, a polymerase chain reaction assay revealed that a B. gibsoni marker gene was intermittently present in peripheral blood after atovaquone therapy, indicating that the organism had not been eliminated, and parasites reappeared in blood smears 33 days after the last treatment. To investigate the change in sensitivity against atovaquone, an in vitro sensitivity test was performed using peripheral blood obtained from an untreated dog that was infected with the original parasite isolate, and from two of the experimentally infected and atovaquone-treated animals (blood was collected at the time of the post-treatment recurrence of the B. gibsoni infection). Atovaquone was added to the culture medium to final concentrations of 0.1, 1, 10, 100, and 1000 nM. For the untreated parasites, complete growth inhibition occurred at 1000 nM of atovaquone, whereas the recurrent parasites were inhibited by only 39.52 +/- 8.34% and 31.31 +/- 8.14% at this concentration after 48 h of incubation. Thus, the recurring parasites were less sensitive to atovaquone than the untreated originally isolated parasites. PMID- 15350658 TI - Neospora caninum infection in breeder bulls: seroprevalence and comparison of serological methods used for diagnosis. AB - AIMS: To investigate the seroprevalence of Neospora caninum infection in breeder bulls. In addition, the level of agreement of serological methods used was evaluated. METHODS: A total of 285 bulls of different ages and breeds raised for reproductive purposes in Spain were sampled for serum antibody activity to N. caninum. Sera were tested using three validated assays: the indirect fluorescent antibody test (IFAT) and two commercially available enzyme-linked immunosorbent assays (ELISA). RESULTS: Antibodies to N. caninum were found in 13.7% of bulls sampled by at least one of the three validated serological assays. The seroprevalence of neosporosis oscillated between 11.2 and 13.3% depending on the serological technique used. No significant associations (P > 0.05) were observed between N. caninum infection and bull breed or age. The serum antibody levels were predominantly low and close to threshold levels in all positive samples. A very good agreement was found among the serological methods used. CONCLUSIONS: This is the first seroprevalence study of bovine neosporosis in breeder bulls and results showed a moderate presence of N. caninum chronic infections. PMID- 15350659 TI - Incidence and prevalence of tick-borne haemoparasites in domestic ruminants in Ghana. AB - Giemsa-stained thin blood smears prepared monthly from cattle, sheep and goats in the Greater Accra region of Ghana between May 1994 and December 1996 were examined for presence of tick-borne haemoparasites. The majority of animals were less than 2 months old at the start of the survey. Monthly and cumulative incidences are presented of Anaplasma sp., Babesia bigemina, Borrelia sp., Eperythrozoon sp., Theileria mutans and Theileria velifera in cattle, Anaplasma sp., Borrelia sp., and Theileria sp. in sheep, and Anaplasma sp. in goats. T. mutans was the commonest parasite in cattle, with 100% incidence in calves by 10 months of age, and Anaplasma was commonest in small ruminants. The relative prevalence of these haemoparasites in blood smears from cattle, sheep and goats sampled on a single occasion at sites in all 10 regions of Ghana was found to be similar, though actual infection rates were lower. Packed cell volume (PCV) measurements from the sampled animals are also presented; no seasonal trends were evident in the PCV of the cattle, sheep and goats sampled monthly. In animals sampled on a single occasion, mean PCV was significantly higher in cattle and sheep without detectable haemoparasite infection, and in cattle was lowest in animals positive for both Babesia and Anaplasma, while there was no difference in mean PCV levels between parasitised and non-parasitised goats. PMID- 15350660 TI - Efficiency of the oral, intramuscular and subcutaneous routes for the experimental infection of hamster and sheep with Schistosoma bovis. AB - The percutaneous administration of cercariae is the usual method for experimental infections with Schistosoma bovis. These procedures are laborious and have important inconveniences when working with a large number of animals, especially if they are ruminants. In the present study, the efficiency of the oral, intramuscular and subcutaneous routes are evaluated by comparison with the percutaneous route in experimental infections with S. bovis. The infections developed in hamsters and sheep were evaluated taking as a basis the parasite burden, the concentrations of eggs in tissues and the levels of anti-Schistosoma antibodies. The oral infection failed in both hamsters and sheep. The administration of the cercariae by the intramuscular route was effective in sheep, developing infections of intensity similar to that of the infections acquired percutaneously. In hamsters, on the contrary, although all the animals developed the infection, they were very little intense. The injection of the cercariae by the subcutaneous route induces acceptable infections in hamsters and can also be an alternative route to percutaneous exposure. The levels of the anti Schistosoma bovis antibodies detected in hamster and sheep were proportional to the number of worms present, which shows that the humoral response is a good indicator of the intensity of the infections. It can be concluded that the intramuscular route is a good alternative to the percutaneous route for experimental infections of sheep with S. bovis. Likewise, the subcutaneous route can also substitute, with some advantages, the percutaneous infections in hamsters. PMID- 15350661 TI - Genomic characterization of lung flukes, Paragonimus heterotremus, P. siamensis, P. harinasutai, P. westermani and P. bangkokensis by RAPD markers. AB - Random amplified polymorphic DNA (RAPD) markers were assayed in an attempt to discriminate among five species of Paragonimus. Genomic DNAs of two strains of Paragonimus heterotremus from two provinces in Thailand, Saraburi and Phitsanulok, as well as of P. siamensis, P. harinasutai, P. westermani and P. bangkokensis were extracted and amplified by an arbitrary primer, namely P2 (5 GTTTCGCTCC-3). RAPD patterns showed that those five species were genetically distinct, although they shared genomic DNA to some extent. This primer could also distinguish between two strains of P. heterotremus. The polymorphism observed allowed to construct a relationship dendrogram. The phylogenetic dendrogram showed that the P. heterotremus strains were closest to P. harinasutai, followed by P. siamensis, P. bangkokensis and P. westermani. PMID- 15350662 TI - Humoral immune response against Fasciola gigantica fatty acid binding protein. AB - Recombinant fatty acid binding protein of Fasciola gigantica was expressed in Escherichia coli and purified by nickel chelating affinity chromatography. The recombinant protein along with native fatty acid binding protein (FABP) isolated from the parasite were evaluated for their potential in the diagnosis of F. gigantica infection in sheep, cattle and buffaloes, both by ELISA and western blotting. Results of this study indicate that there is no humoral immune response generated against this protein in the experimental infection of these ruminants with F. gigantica, thereby limiting the usefulness of this antigen in the early diagnosis of fasciolosis in these animals. Also, the paper discusses the probable reasons for the failure of this protein in detecting humoral response in these animals by ELISA and immunoblotting. PMID- 15350663 TI - Studies using Teladorsagia circumcincta in an in vitro direct challenge method using abomasal tissue explants. AB - An in vitro direct challenge (IVDC) method in which abomasal tissue explants maintained at 37 degrees C in Hanks/Hepes solution (pH 7.6) in a high oxygen concentration (incubator gassed with pure oxygen and sealed prior to use) were challenged with exsheathed third stage larvae of Teladorsagia circumcincta has been used to investigate the tissue association phase of larval establishment. Studies using naive and recently challenged immune sheep and goat abomasal tissues have shown significant reductions (P < 0.05) in the numbers of tissue associated larvae in material from immune animals. The mechanisms that affect the tissue association process appear to be labile since more larvae were recovered from the tissue digests of previously infected sheep that had not been recently exposed to larval challenge in comparison to those that had been recently challenged (P < 0.05). The method has also been used to demonstrate the influence of protein nutrition on the establishment of larvae in abomasal tissues and region specific differences in the efficacy of exclusion mechanisms. The technique appears to have potential as a means of investigating the crucial first phase of the process of establishment. PMID- 15350664 TI - Effect of the antiparasitic drugs fenbendazole and ivermectin on the soil nematode Pristionchus maupasi. AB - Pristionchus maupasi, a soil nematode, was used to elucidate the potential ecotoxic effect of the two anthelmintics fenbendazole and ivermectin in cattle dung. The population growth of P. maupasi was greater in faeces from cattle harbouring active Panacur- or Ivomec-boli, which are releasing fenbendazole and ivermectin to the rumen, respectively, compared to the growth in control faeces. In dose-response experiments it could be shown that the pure chemical compound of fenbendazole was increasingly nematocidal to P. maupasi in concentrations from 10 to 20 microg/g faeces (ww, i.e. wet weight) and the pure compound of ivermectin was effective above 3 microg/g faeces (ww). The results indicate that neither fenbendazole nor ivermectin have any acute toxic effect on P. maupasi in naturally excreted concentrations of the pure drugs, together with their metabolites in faeces from bolus-treated cattle. Both drugs are excreted in concentrations that are non-toxic to P. maupasi. PMID- 15350665 TI - Seroepidemiological study of canine ehrlichial infections in Yamaguchi prefecture and surrounding areas of Japan. AB - Randomly selected serum samples from 150 dogs from Yamaguchi and neighbouring prefectures were subjected to the indirect immunofluorescent assay to detect antibodies against Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia muris and Ehrlichia from Ixodes ovatus. A total of 30 out of the 150 serum samples reacted with at least one of the antigens at a titer of 1:20 or more. Considerable cross reactivity was seen and most samples reacted with at least two different antigens. Fifteen (10.0%) dogs had higher titers to E. canis than any of the other antigens. Four (2.7%) dogs had higher titers to Ehrlichia from Ixodes ovatus and one (0.6%) dog had higher titers to E. muris compared to the other antigens. The findings suggest that these five dogs may be infected with the domestic Ehrlichia of Japan. The remaining ten dogs had similar high titers to two or more of the antigens. This is the first serological evidence obtained of canine infection with the domestic Ehrlichia of Japan. PMID- 15350666 TI - Clinical development and serological antibody responses in sheep and rabbits experimentally infested with Psoroptes ovis and Psoroptes cuniculi. AB - Psoroptes ovis of sheep origin, and Psoroptes cuniculi of rabbit origin were used in experimental infestations. In experiment I, groups of four rabbits and four sheep were infested with 50-100 mites of each isolate on the skin of the back (skin infestation, SI) or in the external auditory canal (aural infestation, AI). In rabbits, SI and AI with P. cuniculi and AI with P. ovis induced in all animals typical ear lesions and pronounced antibody reactions to P. cuniculi antigens in ELISA. After SI of rabbits with P. ovis no clinical signs were detected, no mites could be reisolated and no specific antibodies were detected. In sheep, P. ovis SI induced mange whereas AI did not induce typical clinical signs and mites could not be reisolated. In both these animal groups, ELISA revealed pronounced and comparable specific antibody reactions. After SI and AI with P. cuniculi no clinical symptoms were observed and no mites could be reisolated. Nevertheless, low levels of specific antibody were detected. In experiment II, clinical progression and antibody reactions to P. ovis SI in naive sheep were compared with sheep previously exposed to P. ovis or P. cuniculi. In both pre-exposed groups of animals, clinical signs appeared within 2 days after challenge infestation and three days earlier than in primarily infested sheep. Subsequently, no obvious difference in the clinical progression was observed between the three groups of animals. The results of this study document antigenetic crossreactivity of the two morphologically and genetically distinguishable Psoroptes species but differences in their biological behaviour and virulence which both are of epidemiological and taxonomic relevance. PMID- 15350667 TI - Dirofilaria immitis: worm burden and pulmonary artery proliferation in dogs from Michigan (United States). AB - Despite the ability to prevent heartworm disease, infection with Dirofilaria immitis continues to be a major problem for domestic dogs. To determine worm burden in heartworm-positive dogs from three county animal shelters in the state of Michigan in the United States and to assess the relationship between gross intimal proliferation and worm burden, necropsy was done on 176 heartworm positive dogs. Adult heartworms were found in the heart and pulmonary artery of 170 of the 176 (96.6%) dogs examined. Mean worm burden was 14 +/- 13 (range 0 85). Fifty-nine percent of dogs had < or =10 heartworms. In contrast, 52% of dogs in a published report from the southern US (Florida) had worm burdens >10 [C.H. Courtney, Q.Y. Zeng, The structure of heartworm populations in dogs and cats in Florida, in: Proceedings of the American Heartworm Symposium, 1989]. These data suggest that mean worm burden in northern areas may be < or = that in warmer areas. Also, since diagnostic tests are less sensitive with lower worm burdens, diagnosis of heartworm infection in Michigan and other surrounding more northern states may be a greater challenge than in areas with higher worm burdens. PMID- 15350668 TI - Cytochrome oxidase I gene sequence of Hypoderma sinense infecting yaks in the Qinghai-Tibet high plateau of China. AB - The previous scanning electron microscopy study of the Hypoderma species suggested that Hypoderma sinense Pleske (H. sinense) was different from Hypoderma bovis (H. bovis) and Hypoderma lineatum (H. lineatum). In this study, the mitochondrial cytochrome oxidase I (COI) gene sequence of H. sinense was compared with those of the other two species. The H. sinense sequence showed only 88.3% homology to H. bovis and 88.5% to H. lineatum. The results of the sequencing analysis confirm that H. sinense is a different species from H. bovis and H. lineatum. PMID- 15350669 TI - Correlations of patch test reactivity and the repeated open application test (ROAT)/provocative use test (PUT). AB - The clinical relevance of patch test reactions is often difficult to determine. Use tests have been developed to further evaluate the significance of patch test results. We review studies that have defined correlations between the threshold concentrations at patch testing and the outcome of use tests for particular chemical allergens. Results of patch testing with serial dilutions of colophony, cinnamic aldehyde, and isoeugenol have shown concordance with the outcome of use tests. On the other hand, poor correlations between patch test reactivity and the use test were demonstrated in another study on isoeugenol and on studies on hydroxycitronellal, formaldehyde and chromium. These studies shed light on some factors that may influence the outcome of use tests. Individual factors such as patch test sensitivity, regional variations in reactivity and percutaneous penetration appear to play significant roles. Exposure dose, length of time of exposure, and other factors yet to be determined also affect degree of reactivity. Because patients with low thresholds in serial dilution patch testing are known to react to lower concentrations of products at use testing, results of these studies may be used to help identify subjects with a high risk of developing clinical disease. From the public health standpoint, data obtained may be used as a guide in limiting exposure concentrations in consumer products. PMID- 15350670 TI - Morphological and microarray analysis of T-2 toxin-induced rat fetal brain lesion. AB - To examine morphological and gene expression changes induced by T-2 toxin in the fetal brain in detail, pregnant rats on day 13 of gestation were treated orally with a single dose of T-2 toxin (2 mg/kg) and sacrificed at 1, 3, 6, 9, 12 and 24 h after treatment (HAT). Histopathologically, the number of apoptotic neuroepithelial cells in the telencephalon increased from 1 HAT and peaked at 12 HAT. Based on the histopathological examinations, microarray analysis was performed at 6, 12 and 24 HAT. Microarray analysis showed that the expression of oxidative stress-related genes (heat shock protein 70 (HSP70) and heme oxygenase (HO)) was strongly induced by T-2 toxin at 12 HAT, the peak time point of apoptosis induction. The expression of mitogen-activated protein kinase (MAPK) related genes (MEKK1 and c-jun) and other apoptosis-related genes (caspase-2 and insulin-like growth factor-binding protein-3 (IGF-BP3)) was also induced by the T 2 toxin treatment. The changes observed by microarray analysis were confirmed for four up-regulated genes (HSP70, HO, IGF-BP3 and VEGF-A) using real-time RT-PCR. Our results suggest that the T-2 toxin-induced apoptosis in the fetal brain is due to oxidative stress, and that the MAPK pathway may be involved in T-2 toxin induced toxicity. PMID- 15350671 TI - Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms: effects on rat metabolism and gut histology. AB - This present work was undertaken to answer two basic questions (a) is C. argentea lectin part of the general defensive strategy of the plant against predation by animals? (b) if so, how does it act on them? To achieve these goals the lectin from C. argentea seeds was purified to homogeneity and included at a 2% level in a diet containing 10% total protein and given to growing rats for 10 days. In vivo it was noted that the lectin from C. argentea is resistant to gut proteolysis, binds to the cells lining the small intestine and induces enlargement in the small intestine, caecum and colon, kidneys and pancreas compared to control rats exposed to the egg-white diet (EW). As the diet containing the purified C. argentea lectin has the same basic composition and protein content of EW diet, the small intestine, kidney and pancreas enlargements are clearly lectin-specific effects. Moreover the animals exposed to the lectin containing diet presented a significant reduction in the growth rate and lower values of digestibility, NPU and biological value compared to animals fed on a control lectin-free diet. Thus the data from this present study and the report that the C. argentea lectin has insecticidal activity upon Callosobruchus maculatus larvae which attacks cowpea (Vigna unguiculata) seeds reinforce the hypothesis that lectins take part in the mechanisms against herbivory. PMID- 15350672 TI - Effects of dietary supplements on induction and inhibition of cytochrome P450s protein expression in rats. AB - Recent surveys show that 18% of adults in the United States use prescription drugs concurrently with herbal or vitamin dietary products. Despite possible dietary supplement-drug interactions through modulation of cytochrome P450s (CYPs), dietary supplements have not been studied at a screening scale to assess their effects on CYPs. In this study, 116 herbal dietary supplements, commercially available for nutrient supply and weight management, were administered to rats to test whether they modulate the expressions of CYP1A2, 2C11, 2D1, 2E1 and 3A1 proteins. Seventy-five percent of the 116 dietary supplements modulated at least one CYP, while 25% had no effect. CYP2C11 protein expression was the most inhibited by supplements (51%), whereas CYP1A2, 2D1, 2E1 and 3A1 were the least inhibited (12-18%). CYP1A2 was the most induced, modulated by 21% supplements, while CYP2E1 and 3A1 were moderately induced (7-8%). CYP2C11 and 2D1 were not induced by any supplement tested in this study. Thus, this study suggests that dietary supplement-drug interactions may occur through modulation of CYPs in humans when they are taken simultaneously. PMID- 15350673 TI - Toxicology and carcinogenesis studies of microencapsulated trans-cinnamaldehyde in rats and mice. AB - trans-Cinnamaldehyde is a widely used natural ingredient that is added to foods and cosmetics as a flavoring and fragrance agent. Male and female F344/N rats and B6C3F(1) mice were exposed to microencapsulated trans-cinnamaldehyde in the feed for three months or two years. All studies included untreated and vehicle control groups. In the three-month studies, rats and mice were given diets containing 4100, 8200, 16,500, or 33,000 ppm trans-cinnamaldehyde. In rats, feed consumption was reduced in all exposed groups. In mice, feed consumption was reduced in the highest dose groups. Body weights of all treated males were less than controls. Body weights were reduced in female rats exposed to 16,500 or 33,000 ppm and female mice exposed to 8200 ppm or greater. All rats survived to the end of the study but some male mice in the highest dose groups died due to inanition from unpalatability of the dosed feed. The incidence of squamous epithelial hyperplasia of the forestomach was significantly increased in rats exposed to 8200 ppm or greater and female mice exposed to 33,000 ppm. In mice, the incidence of olfactory epithelial degeneration of the nasal cavity was significantly increased in males and females exposed to 16,500 ppm and females exposed to 33,000 ppm. In the two-year studies, rats and mice were exposed to 1000, 2100, or 4100 ppm trans-cinnamaldehyde. Body weights were reduced in mice exposed to 2100 ppm and in rats and mice exposed to 4100 ppm. In rats, hippuric acid excretion was dose proportional indicating that absorption, metabolism, and excretion were not saturated. No neoplasms were attributed to trans-cinnamaldehyde in rats or mice. Squamous cell papillomas and carcinomas of the forestomach were observed in male and female mice but the incidences were within the NTP historical control range and were not considered to be related to trans-cinnamaldehyde exposure. PMID- 15350674 TI - Genotoxicity testing of a fenugreek extract. AB - Fenugreek seeds have been used in traditional medicines as a remedy for diabetes. Rich in protein, fenugreek seeds contain the unique major free amino acid 4 hydroxyisoleucine (4-OH-Ile), which has been characterized as one of the active ingredients for blood glucose control. Current use of fenugreek in foodstuff has been limited to its role as a flavoring agent, and not as an ingredient to help mitigate the blood glucose response for people with diabetes. As part of a safety evaluation of novel ingredients for use in blood glucose control, the potential genotoxicity of a fenugreek seed extract (THL), containing a minimum of 40% 4-OH ILE, was evaluated using the standard battery of tests (reverse mutation assay; mouse lymphoma forward mutation assay; mouse micronucleus assay) recommended by US Food and Drug Administration (FDA) for food ingredients. THL was determined not to be genotoxic under the conditions of the tested genetic toxicity battery. The negative assay results provide support that addition of THL to foodstuffs formulated for people with diabetes is expected to be safe. A wide safety margin is established, as anticipated doses are small compared to the doses administered in the assays. PMID- 15350675 TI - Effects of glycolic acid on the induction of apoptosis via caspase-3 activation in human leukemia cell line (HL-60). AB - Apoptosis is a particular process that leads to the programmed cell death, and it has been a potentially therapeutic target of cancer. In this study, we evaluated the possible apoptotic effects of glycolic acid on human leukemia cell line (HL 60) in vitro. The morphological changes, cell viability, apoptosis induction, and caspase-3 activity were measured by phase microscopy, flow cytometry, and Western blot analysis. Morphological changes including shrinkage of cells were clearly demonstrated in HL-60 cells treated with increasing concentrations of glycolic acid. Cell viability was significantly affected by glycolic acid treatment in a dose- and time-dependent manner. In comparison to the control group, glycolic acid treatment had a profound effect in the induction of apoptosis by flow cytometric assays. In the cell cycle analysis, glycolic acid caused the increased percentage of cells in G2/M phase and the decreased expression of the cyclin A and cyclin B1, suggesting the induction of G2/M arrest of cell cycle by glycolic acid. Moreover, glycolic acid treatment promoted caspase-9 and -3 activity in a dose-dependent manner, but caspse-8 activity was not affected during the same process. Glycolic acid co-administrated with broad-spectrum caspase inhibitor, z VAD-fmk, caspase-3 activity was blunted and apoptosis was also markedly blocked in HL-60 cells. In conclusion, glycolic acid-induced apoptosis in HL-60 cells may be through the activation of caspase-3. Future studies focusing on cell signaling and biological significance of glycolic acid-induced apoptosis would lead to exploring the mechanisms of chemotherapeutic potency of glycolic acid in human cancers. PMID- 15350676 TI - Effects of amitraz on cytochrome P450-dependent monooxygenases and estrogenic activity in MCF-7 human breast cancer cells and immature female rats. AB - This study investigated the ability of amitraz, a formamidine insecticide, to induce cytochrome P450-dependent monooxygenases and to disrupt estrogenic activity in human breast cancer MCF-7 cells and immature female rats. In MCF-7 cells, treatment with 10 microM amitraz for 24 h increased 7-ethoxyresorufin O deethylase activity in cell homogenate. Treatment of MCF-7 cells with 1 and 10 microM amitraz for 3 h replaced previously bound [(3)H]17beta-estradiol (E(2)) from estrogen receptors. Treatment with 0.1 and 1 microM amitraz for 2 days inhibited [(3)H]thymidine incorporation into the DNA of MCF-7 cells while the inhibition was blocked in cells co-treated with 1 nM E(2) and amitraz. In immature female rats, treatment with 50 mg/kg amitraz intraperitoneally for 3 days increased cytochrome P450 content, 7-ethoxyresorufin, methoxyresorufin and pentoxyresorufin O-dealkylases, and benzo[a]pyrene hydroxylase activities in liver microsomes. The results of immunoblot analysis revealed that amitraz induced liver microsomal CYP1A1/2, 2B1/2B2, and 3A proteins. Treatment with 10 and 25 mg/kg amitraz for 3 days dose-dependently decreased uterine weight and peroxidase activity in immature female rats while the decreases were blocked in rats co-treated with 10 microg/kg E(2) and 10 or 25 mg/kg amitraz. These in vitro and in vivo findings suggest that amitraz induces multiple forms of P450 and exerts weak antiestrogenic activity. PMID- 15350677 TI - Liver carcinogenesis and formation of 8-hydroxy-deoxyguanosine in C3H/HeN mice by oxidized dietary oils containing carcinogenic dicarbonyl compounds. AB - Oxidized dietary oils (lard, soybean oil, and sardine oil) were orally administered to C3H/HeN male mice. After 6 months, benign hepatocellular adenoma was observed in the mice treated with all three oxidized dietary oils. After 12 months, malignant hepatocellular carcinoma and hepatoblastoma were observed in addition to the benign tumor. Oxidized sardine oil caused the highest tumor incidence (35%) and malignant tumors (27.5%) among the oxidized dietary oils tested. Mice treated with oxidized lard and sardine oil exhibited a significant increase of 8-OH-dG in the livers. The amounts of 8-OH-dG found in the mice treated with oxidized sardine oil correlated with the rates of tumor incidence. After 6 months, mRNA decreased in the case of oxidized lard and sardine oil, whereas it increased in the case of oxidized soybean oil, either in 8-oxoguanine DNA glycosylase (OGG1) or in 8-oxo-dGTPase. On the other hand, there was no appreciable change in mRNA, in either OGG1 or 8-oxo-dGTPase, after 12 months. Oxidized sardine oil contained the highest level of malonaldehyde (MA) (713+/ 91.1 nmol/g) and glyoxal (33.3+/-5.2 nmol/g) among three oxidized oils. The malignant tumor incidence correlated with the high level of MA and glyoxal found in the dietary oils tested. PMID- 15350678 TI - Evaluation of the association of demographics and beverage consumption with dental caries. AB - There is much speculation about the role of carbonated soft drinks in the development of dental caries. We examine the relationships between certain demographic variables, beverage consumption, and professional dental care and their contribution to the number of decayed, missing, and filled surfaces (DMFS). The National Health and Nutrition Examination Survey III (NHANES III) conducted by the National Center for Health Statistics was used to examine DMFS among four age groups: 17-24 yr, 25-40 yr, 41-60 yr, and over 60 yr. Age has a strong, positive relationship with DMFS. About 15% of young adults aged 17-24 yr have no DMFS, and the average DMFS is only 10.4 in this age group. Less than 1% of adults over age 40 have a DMFS score of 0, and nearly one in five has the maximum of 128 DMFS. Controlling for other factors, whites have more DMFS than do blacks and Hispanics. Beverages have a weak, but statistically significant, relationship with DMFS among the older age groups only. Regular professional dental care was associated with lower DMFS among the older age groups. Age and ethnicity are the strongest predictors of DMFS. Young adults have fewer DMFS regardless of dietary factors. This study suggests that carbonated soft drinks are not associated with poor dental health. Useful strategies to reduce dental caries involve good personal dental hygiene, regular use of fluoridated toothpastes and mouthwashes, and regular care by dental professionals. PMID- 15350679 TI - 28-Day repeated dose toxicity study of dried microorganism in rats. AB - Ubidecarenone, also known as CoQ(10), is currently sold as a dietary supplement in the United States, with a majority of these products derived from the fermentation of carbohydrates or tobacco leaf extracts. In addition to its availability in dietary supplements, CoQ(10) is now being considered for use in foods. Accordingly, as part of the process for attaining "Generally Recognized as Safe" status, and to supplement information already available regarding the safety of CoQ(10) per se, a 28-day oral toxicity study in rats was conducted to evaluate the subacute safety of a microorganism biomass used as a new source in CoQ(10) production. Groups of Crj:CD(SD) rats (SPF) (6 males or females per group, 4 groups per sex) received dried microorganism at doses of 0, 500, 1000 or 2000 mg/kg/day via intragastric intubation. Clinical observations were recorded, and body weight, and food and water consumptions measured throughout the study. At the end of the study, aortic blood samples were collected from all animals for analysis of hematological and clinical chemistry parameters, and gross pathologic examination was performed. Histopathologic examination was performed on select tissues from the control and high-dose groups. There were no treatment-related changes that were considered to be of toxicological significance. Since rats treated with 2000 mg/kg of dried microorganism did not demonstrate any treatment related changes, the no-observable-adverse-effect level (NOAEL) for dried microorganism was estimated to be greater than 2000 mg/kg/day under the present study conditions. PMID- 15350680 TI - Application of adsorbent agents technology in the removal of aflatoxin B(1) and fumonisin B(1) from malt extract. AB - The commercially hydrated sodium calcium aluminosilicate (HSCAS) and the Egyptian montmorillonite (EM) had an excellent capability of adsorbing AFB(1) and FB(1) in an aqueous solution at different tested levels. The adsorption ratio of HSCAS ranged from 95.3% to 99.1% and 84.7% to 92.4% of the available AFB(1) and FB(1) respectively. EM showed an adsorption ratio ranged from 95.4% to 99.2% and 78.2% to 92.2% for AFB(1) and FB(1) respectively. Both adsorbents were effective at 0.5% level. Results of the ability of these adsorbents at level of 0.5% (w/v) to adsorb AFB(1) and FB(1) in malt extract spiked with 50, 100 and 200 ppb indicated that the capability of adsorbing of HSCAS ranged from 98.5% to 98.9% and 88.2% to 91.9% for AFB(1) and FB(1) respectively. Whereas, the capability of adsorbing of EM ranged from 98.1% to 98.7% and 88.2% to 92.5% for AFB(1) and FB(1) respectively. PMID- 15350681 TI - Chemical characterization of different sugar-casein Maillard reaction products and protective effects on chemical-induced cytotoxicity of Caco-2 cells. AB - This study examined the chemical modification of casein with three monosaccharides [e.g., glucose (Glc), fructose (fru), and ribose (Rib)], following prolonged gentle heating to product the Maillard reaction (MR). Changes in chemical modification of casein in different sugar-casein MR was monitored using temporal patters of change in fluorescence development, UV absorbance and casein molecular mass, which was studied using SDS-PAGE and MALDI-TOF mass spectrometry. Rib-casein MRP exhibited significantly (p<0.05) advanced fluorescence development at three days, compared to Glc- and Fru-casein MRPs, which corresponded to marked (p<0.05) differences in greater UV absorption of Rib casein MRPs later on after 15 days of processing. These results were associated with the generation of different high molecular weight complexes generated from the Rib-casein MR, compared to Glu- and Fru-casein MRPs, respectively. Antioxidant activity of all three sugar-modified caseins was also assessed when cultured with Caco-2 cells. A significant, but similar protective effect against 2,2'-azobis-(2-amidinopropane)dihydrochloride (AAPH) radical-induced cytotoxicity, and ferrous- and cupric-induced toxicity of Caco-2 cells was observed for all three sugar-casein MRPs. However, these protective effects were not significantly different among the sugar-modified caseins and native casein. The results suggest that protection of Caco-2 cells from free radical-ferrous- and cupric-induced cytotoxicity by casein was not altered by modification by MR, regardless of source of monosaccharide involved in the reaction. PMID- 15350682 TI - Lack of spermatotoxic effects of methyl and ethyl esters of p-hydroxybenzoic acid in rats. AB - Parabens are alkyl esters of p-hydroxybenzoic acid widely used as preservatives in foodstuffs, cosmetics toiletries and pharmaceuticals. These compounds are known to exert a weak estrogenic activity in estrogen receptor assays in vitro. In addition butyl and propyl parabens show uterotrophic activity in vivo. It was previously shown that exposure of post-weaning rats and mice to butyl or propyl parabens adversely affects the secretion of testosterone and the function of the male reproductive system. In the present study, it is shown that methyl and ethyl parabens do not adversely affect the secretion of sex hormones or the male reproductive function. Methyl and ethyl parabens were administered to 25-27-day old rats assigned to five groups of eight animals each, at doses of 0.1% and 1.0% each in the rat's diet. At the end of 8 weeks, the rats were sacrificed by decapitation and the weights of the testes, epididymides, prostates, seminal vesicles and preputial glands were determined. There were no treatment-related effects of either compound on the organ weights in any of the study groups. Neither compound exhibited anti-spermatogenic effects nor elicited changes in levels of testosterone, LH and FSH at a dose level of about 1000 mg/kg of body weight per day. PMID- 15350683 TI - Identification and reduction of urinary aflatoxin metabolites in dogs. AB - Hydrated sodium calcium aluminosilicate (HSCAS) is a phyllosilicate clay commonly used as an anticaking agent in animal feeds. HSCAS tightly and selectively adsorbs aflatoxin. In 1998, 55 dogs died in Texas after eating dog food containing aflatoxin (150-300 ppb). The corn in the diets was contaminated with aflatoxin. Six dogs were given a low-level, sub-clinical dose of aflatoxin B(1). On average, 71.5% of aflatoxin M(1) cleared within 6 h after dosing, increasing to 90.4% after 12 h. Aflatoxin M(1) was no longer detectable in urine after 48 h. Aflatoxin P(1) was not found in urine compared to large amounts of M(1) and trace amounts of Q(1). In a crossover study, six dogs randomly fed a commercial dog food (no-clay control) or coated with HSCAS (0.5% by weight) were subsequently administered a sub-clinical dose of aflatoxin B(1). Diets were switched and the process repeated. The HSCAS-coated diet significantly reduced urinary aflatoxin M(1) by 48.4%+/-16.6 SD versus the control diet. In conclusion, HSCAS protects dogs fed diets with even minimal aflatoxin contamination. Despite regular and careful ingredient screening for aflatoxin, low concentrations may reach the final product undetected. Therefore, HSCAS may provide the pet food industry further assurance of canine diet safety. PMID- 15350684 TI - Thresholds of elicitation depend on induction conditions. Could low level exposure induce sub-clinical allergic states that are only elicited under the severe conditions of clinical diagnosis? AB - While numerous studies have examined dose/response relationships occurring in the experimental induction of contact allergic dermatitis, fewer have examined the effects of varying the doses of both induction and challenge. Recently published studies have however done this and they all show the same remarkable observation: the threshold of elicitation decreases as the doses used to induce the allergy increase. This has important implications. One is that it may be more complicated to determine clear threshold doses below which allergic responses are not seen. It is also proposed that normal exposure to weak allergens such as some fragrance materials may induce "sub-clinical" allergic states which will not be elicited under these same exposure conditions but which may become apparent under the more severe conditions of clinical diagnosis. This may explain why the prevalence of Patch test reactions to some fragrance materials is apparently increasing in the absence of any clearly documented "epidemic" of consumer complaints. PMID- 15350685 TI - Safety evaluation of a triterpenoid-rich extract from bamboo shavings. AB - Triterpenoids, which may have significant application to the development of natural medicines and functional foods as biological active components, are widely distributed throughout the plant kingdom. This paper evaluated the safety of a triterpenoid-rich extract of bamboo shavings (EBS) systematically. (i) Acute toxicity test: The oral maximum tolerated dose of EBS was more than 10 g/kg body weight both in rats and in mice, due to the absence of toxicity according to the criteria of acute toxic classifications. (ii) Mutagenicity test: It had no mutagenicity judged by negative experimental results of Ames test, mouse bone marrow cell micronucleus test and mouse sperm abnormality test. (iii) 30 days feeding study: No abnormal symptoms and clinical signs or deaths had been found in rats in each group during the test. No significant difference had been found in body weight, food consumption and food availability of rats in each test group (P>0.05). In addition, no significant differences were found in each hematology value, clinical chemistry value and organ/body weight ratio, either (P>0.05). No abnormality of any organ was found during histopathological examination. It can be concluded that the extract of bamboo shavings is of low toxicity and support the use of EBS for various foods. PMID- 15350686 TI - Dithiocarbamates residues in Brazilian food and the potential risk for consumers. AB - Dithiocarbamates are a non-systemic group of pesticides widely used to protect crops from fungal diseases. The current methodology used by monitoring laboratories to determine dithiocarbamates in food involves the analysis of CS(2) generated after hydrolysis of the compound present in the sample. This method does not identify the origin of the CS(2) detected, which may or may not be related to the presence of pesticides leading to a potential overestimation of the dietary dithiocarbamate intake. In this paper, 520 food samples (papaya, banana, apple, strawberry, orange, potato, tomato, rice and dry beans) collected in the local market of the Federal District, Brazil, were analyzed for dithiocarbamate content. Detectable levels (> or =10.10 mg/kg CS(2)) were found in 60.8% of the samples, with the highest levels (up to 3.8 mg/kg) found in strawberry, papaya and banana. No residues were found in rice (polished) and only one dry bean sample had detectable levels of the fungicides. Detectable residues were found in the pulp of banana, papaya (including the seeds) and orange (50-62% of the analyzed samples). An exposure assessment, based on dithiocarbamate levels detected in the food crops analyzed in this study, confirms that the intake of dithiocarbamates through food consumption in the country does not represent a health risk to consumers, i.e., the estimated daily intake is less than the acceptable daily intake. Furthermore, the implementation of more selective methodologies to individually analyze these compounds in food monitoring programs in Brazil is not necessary. PMID- 15350687 TI - Urinary acetylated metabolites and N-acetyltransferase-2 genotype in human subjects treated with a para-phenylenediamine-containing oxidative hair dye. AB - In the organism of mammals, important detoxification pathways of arylamines are catalysed by N-acetyltransferase 2 (NAT2). A recent case-control epidemiology study suggested that human NAT2 slow acetylators exposed to oxidative hair dyes may be at greater risk to develop bladder cancer. We therefore profiled urinary [(14)C]-metabolites and NAT2 genotype in eight human subjects following treatment with a dark-shade oxidative hair dye containing [(14)C]-para-phenylenediamine (PPD). Genotyping identified three subjects as slow, and five subjects as intermediate NAT2 acetylators. Within 24 h after treatment, the study subjects excreted a mean total of 0.43+/-0.24% of the applied [(14)C] in the urine, where five different metabolites were found. The major urinary metabolites were concluded to be N-mono-acetylated and N,N'-diacetylated PPD. They were present in all urine samples and amounted to 80-95% of the total urinary [(14)C]. Another metabolite, possibly a glucuronic acid conjugate, was found in 6/8 urine samples at 5-13% of the total urinary [(14)C]. All metabolites appeared to be related to PPD, no evidence of the presence of high-molecular weight dye-intermediates or corresponding metabolites was found. The metabolite profile in the study subjects showed no significant differences between the NAT2 intermediate and NAT2 slow acetylator subgroups. Urine of NAT2 slow acetylators contained N-mono-acetylated PPD at 42.2+/-10.2% and N,N'-di-acetylated-PPD at 54.1+/-7.6% of total urinary radioactivity, while the corresponding values of intermediate acetylators were 46.0+/-8.9% and 45.7+/-9.9%, respectively. Overall, our results suggest that the human acetylation rate of PPD after topical application is independent of the NAT2 genotype status, most likely due to metabolism by epidermal NAT1 prior to systemic absorption. PMID- 15350688 TI - Estimates of trace metal bioavailability to humans ingesting contaminated oysters. AB - Oysters, as very popular food items in France, are subject to rules concerning the maximum acceptable contents of trace metals in foods. The food standards for the quantities of metals permitted are always based on total metal concentrations, and not on the metal concentrations that are potentially bioavailable to the consumer (man). In order to estimate the percentages of accumulated trace metals (i.e. Ag, Cd, Cu and Zn) that are potentially bioavailable during consumption, we have used a simple chemical digestion simulation on the insoluble fraction of oysters. These quantities have been added to the soluble fraction, assuming that metals in this fraction are completely bioavailable. Our experiments were conducted on oysters Crassostrea gigas sampled from five sites on the French Atlantic coast and on oysters Ostrea edulis sampled from Restronguet Creek in the United Kingdom. These sites are characterised by various degrees of metal contamination. This study has allowed us to gain a better estimate of the real concentrations of metals bioavailable to the consumer. Only a part of the total metal present is bioavailable: 36-68% for silver, 44-75% for cadmium, 26-80% for copper and 50-80% for zinc. These new estimates have the potential to contribute to any re-evaluation of the food standards for metals. PMID- 15350689 TI - SPAG11/isoform HE2C, an atypical anionic beta-defensin-like peptide. AB - A human caput epididymidal cDNA, HE2C, was cloned based on its homology to the known chimpanzee counterpart, suggesting that the encoded beta-defensin-like peptide represented a conserved component of the innate epididymidal epithelial defense system in primates. An approximately 6kDa HE2- related peptide was co purified together with other HE2 isoforms from human seminal plasma by affinity chromatography. By its antibody reactivity as shown by Western blot analysis, this peptide was distinct from the more abundant HE2 isoforms and was concluded to correspond to HE2C. Similar to other HE2-encoded isoforms, the endogenous HE2C was proteolytically processed from a larger precursor by a furin-like prohormone convertase. This was confirmed by N-terminal sequencing. In order to study the structural and functional properties of HE2C it was recombinantly expressed in insect cells. Post-translational processing also occurred within these cells, yielding the mature processed HE2C peptide. Correct disulfide bonding of the recHE2C peptide was shown by p-aminophenylarsineoxide(PAPAO)-agarose binding assay. Purified recHE2C strongly bound to Escherichia coli DH5alpha and Bacillus subtilis; however, it did not exhibit microbicidal activity when tested in a radial diffusion assay against these bacteria. Different from the previously described beta-defensins, the mature HE2C peptide has an anionic pI and an algebraic net charge of -1. Also, it lacks the amphipathic transitions, which, according to the Shai-Matzusaki-Huang model, are prerequisite for the membranolytic activity of antimicrobial peptides. PMID- 15350690 TI - A non-specific lipid transfer protein with antifungal and antibacterial activities from the mung bean. AB - A non-specific lipid transfer peptide (nsLTP) with antimicrobial activity was isolated from the mung bean (Phaseolus mungo) seeds. The procedure entailed aqueous extraction, ion exchange chromatography on CM-Sephadex and high performance liquid chromatography (HPLC) on POROS-HS-20. The peptide exhibited a molecular mass of 9.03 kDa in mass spectrometry. It exerted antifungal action toward Fusarium solani, Fusarium oxysporum, Pythium aphanidermatum and Sclerotium rolfsii, and antibacterial action against Staphylococcus aureus but not against Salmonella typhimurium. The lipid binding of this peptide was very similar to that of a previously described lipid transfer protein extracted from wheat seeds and maize seeds, indicating that it possessed lipid transfer activity. The present findings add to the scarcity of the literature on leguminous nsLTPs. PMID- 15350691 TI - Long-sarafotoxins: characterization of a new family of endothelin-like peptides. AB - Sarafotoxins (SRTXs) constitute a family of vasoactive peptides that were initially isolated from the venom of Atractaspis engaddensis, and that are structurally and functionally related to endothelins (ETs). Analysis of the venom of Atractaspis microlepidota microlepidota revealed several new SRTX molecules manifesting some new structural and functional characteristics. These novel SRTXs are longer by three amino acids than the previously described SRTXs, and are designated here "long-SRTXs". Six isoforms, derived from new poly-cistronic precursors, have been identified so far in the venom of this snake. One of these isoforms, designated SRTX-m, was chemically synthesized and its biological properties were studied. Our results show that SRTX-m induces toxicity in mice, mostly due to vasoconstriction, and also that it has a lower toxicity and potency than the more potent SRTX described up to now: sarafotoxin-b (SRTX-b) from A. engaddensis. PMID- 15350692 TI - Effects of peptide molecular mass and PEG chain length on the vasoreactivity of VIP and PACAP(1-38) in pegylated phospholipid micelles. AB - Bioactive properties of certain amphipathic peptides are amplified when self associated with sterically stabilized micelles (SSM) composed of polyethylene glycol (PEG)-conjugated phospholipids. The purpose of this study was to determine the effects of amphipathic peptide molecular mass and PEG chain length on vasoreactivity evoked by vasoactive intestinal peptide (VIP), a 28-amino acid neuropeptide, and pituitary adenylate cyclase-activating peptide(1-38) (PACAP(1 38)), a 38-amino acid neuropeptide, associated with PEGylated phospholipid micelles in vivo. Both peptides were incubated for 2 h with SSM composed of PEG with molecular mass of 2000 or 5000 grafted onto distearoyl phosphatidylethanolamine (DSPE-PEG2000 or DSPE-PEG5000) before use. We found that regardless of peptide molecular mass, PEG chain length had no significant effects on peptide-SSM interactions. Using intravital microscopy, VIP associated with DSPE-PEG5000 SSM or DSPE-PEG2000 SSM incubated at 25 degrees C evoked similar vasodilation in the intact hamster cheek pouch microcirculation. Likewise, PACAP(1-38)-induced vasodilation was PEG chain length-independent. However, SSM associated PACAP(1-38) evoked significantly smaller vasodilation than that evoked by SSM-associated VIP (P < 0.05) at 25 degrees C. When the incubation temperature was increased to 37 degrees C, SSM-associated PACAP(1-38)-induced vasodilation was now similar to that of SSM-associated VIP. This response was associated with a corresponding increase in alpha-helix content of both peptides in the presence of phospholipids. Collectively, these data indicate that for a larger amphipathic peptide, such as PACAP(1-38), greater kinetic energy or longer incubation period is required to optimize peptide-SSM interactions and amplify peptide bioactivity in vivo. PMID- 15350693 TI - In vitro circadian ANP secretion by gene transferring cells encapsulated in polycaprolactone tubes: gene chronotherapy. AB - A new insofar as chronobiologic therapeutic approach by atrial natriuretic peptide (ANP) for hypertension and/or congestive heart failure (CHF) is based on the release of ANP from ANP cDNA transfected Chinese Hamster Ovary (CHO) cells encapsulated in polycaprolactone (PCL) tubes. ANP secretion was maintained for at least 6 months. The encapsulated cells remained viable during culturing. Control cells without transferred ANP cDNA were negative. ANP secretion is circadian periodic, peaking around 04:18, shifted to around 07:56 by melatonin treatment. The encapsulation technique, based on principles of chronotherapy, may provide a more efficient gene therapy, applicable for eventual human implantation of gene transferred cells. PMID- 15350694 TI - Ghrelin enhances the growth of cultured human adrenal zona glomerulosa cells by exerting MAPK-mediated proliferogenic and antiapoptotic effects. AB - Ghrelin is an endogenous ligand of the growth hormone secretagogue receptor (GHS R), two subtypes of which have been identified and named GHS-R1a and GHS-R1b. Evidence has been provided that ghrelin and its receptors are expressed in the adrenal gland, and we have investigated the possible role of the ghrelin system in the functional regulation of the human adrenal cortex. Reverse transcription polymerase chain reaction detected the expression of both subtypes of GHS-Rs exclusively in the zona glomerulosa (ZG). Ghrelin did not significantly affect either basal or agonist-stimulated aldosterone secretion from cultured ZG cells. In contrast, ghrelin raised proliferative activity and decreased apoptotic deletion rate of ZG cells, the maximal effective concentration being 10(-8) M. The growth effects of 10(-8) M ghrelin on cultured ZG cells were not affected by either the protein kinase (PK)A and PKC antagonists H-89 and calphostin-C or the mitogen-activated PK (MAPK) p38 antagonist SB-293580, but were abolished by both the tyrosine kinase (TK) and MAPK p42/p44 antagonists tyrphostin-23 (10(-5) M) and PD-98059 (10(-4) M), respectively. Ghrelin (10(-8) M) enhanced TK and MAPK p42/p44 activities of ZG cells. Preincubation with 10(-5) M tyrphostin-23 blocked the ghrelin-induced stimulation of both TK and MAPK p42/p44, while preincubation with 10(-4) M PD-98059 only annulled MAPK p42/p44 stimulation. Collectively, our findings allow us to conclude that ghrelin, acting via GHS-Rs exclusively located in the ZG, enhances the growth of human adrenal cortex, through a mechanism involving the activation of the TK-dependent MAPK p42/p44 cascade. PMID- 15350695 TI - The melanocortin agonist melanotan II increases insulin sensitivity in OLETF rats. AB - Effects of peripheral administration of melanotan II (MTII), a melanocortin agonist, on insulin sensitivity and glucose tolerance were examined in Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Subcutaneous administration of MTII with osmotic mini-pumps decreased food intake and body weight in OLETF rats. MTII group showed more sensitivity to insulin compared with that allowed to eat ad libitum or pair-fed group in insulin tolerance tests on day 9. MTII group also showed significantly lower glucose values than ad libitum group in glucose tolerance tests on days 11 and 23. Thus, MTII increased insulin sensitivity and improved glucose tolerance in OLETF rats. PMID- 15350696 TI - Galanin inhibits gastric acid secretion through a somatostatin-independent mechanism in mice. AB - The role of somatostatin in galanin-induced inhibition of gastric acid secretion in urethane-anesthetized mice was investigated by using immunoneutralization of endogenous somatostatin and somatostatin receptor type 2 (SSTR2) knockout mice. Intravenous galanin (10 and 20 microg/kg/h) inhibited pentagastrin-stimulated gastric acid secretion by 47 and 33%, respectively. Somatostatin antibody injected i.v. increased acid secretion by 3.5-fold over basal levels but did not modify the antisecretory effects of galanin. Urethane-anesthetized SSTR2 knockout mice had a basal secretion 14-fold higher than wild-type animals, that was inhibited by galanin (10 and 20 microg/kg/h) by 49 and 31% respectively. In mice galanin inhibits gastric acid secretion through a somatostatin-independent mechanism. PMID- 15350697 TI - Circadian phase difference of leptin in android versus gynoid obesity. AB - A circadian rhythm in serum leptin, measured every 4 h for 24 h, characterizes normal-weight women (N = 14), and women with gynoid (N = 17) or android (N = 26) obesity, peaking around midnight (P < 0.05), but differing by about 3 h between android and gynoid women (P < 0.01). Obesity is associated with a higher MESOR (rhythm-adjusted mean; P < 0.001) and a smaller relative circadian amplitude (P < 0.05). Gynoid obesity is associated with a larger circadian amplitude of cortisol (P < 0.05), whereas android obesity is associated with a larger circadian amplitude and a higher MESOR of insulin (P < 0.05). Understanding putative mechanisms underlying different body fat distribution may lead to improved chronotherapeutic measures. PMID- 15350698 TI - Ultrastructure of orexin-1 receptor immunoreactivities in the spinal cord dorsal horn. AB - The ultrastructural properties of orexin 1-receptor-like immunoreactive (OX1R-LI) neurons in the dorsal horn of the rat spinal cord were examined using light and electron microscopy techniques. At the light microscopy level, the most heavily immunostained OX1R-LI neurons were found in the ventral horn of the spinal cord, while some immunostained profiles, including nerve fibers and small neurons, were also found in the dorsal horn. At the electron microscopy level, OX1R-LI perikarya were identified containing numerous dense-cored vesicles which were more heavily immunostained than any other organelles. Similar vesicles were also found within the axon terminals of the OX1R-LI neurons. The perikarya and dendrites of some of the OX1R-LI neurons could be seen receiving synapses from immunonegative axon terminals. These synapses were found mostly asymmetric in shape. Occasionally, some OX1R-LI axon terminals were found making synapses on dendrites that were OX1R-LI in some cases and immunonegative in others. The synapses made by OX1R-LI axon terminals were found both asymmetric and symmetric in appearance. The results provide solid morphological evidence that OX1R is transported in the dense-cored vesicles from the perikarya to axon terminals and that OX1R-LI neurons in the dorsal horn of the spinal cord have complex synaptic relationships both with other OX1R-LI neurons as well as other neuron types. PMID- 15350699 TI - Vascular and brain neuropeptide Y in banded and spontaneously hypertensive rats. AB - Debate exists regarding the relative importance of neuropeptide Y (NPY) in the pathogenesis of genetic and non-genetic hypertension. NPY concentrations were compared in conduit, mesenteric and renal vasculatures and in hypothalamic and medullary regions of age-matched normotensive control, aortic banded and spontaneously hypertensive rats (SHRs). Lower NPY concentrations were measured in the pre-optic area of banded rats compared to controls and SHR. Renal vein NPY levels were reduced in banded animals, whereas renal artery levels were decreased in SHR. In mesenteric arteries, NPY concentration was selectively increased in SHR. These findings suggest that local hemodynamic alterations influence endogenous levels of this potent vasoconstrictor. PMID- 15350700 TI - Adrenomedullin as a sensitive marker for coronary and peripheral arterial complications in patients with atherosclerotic risks. AB - Plasma adrenomedullin (AM) levels are elevated in various pathological states including cardiovascular and inflammatory diseases. The present study investigated whether an increased AM level is a marker of vascular complications in patients with atherosclerotic risks. In 114 patients with cardiovascular risks and/or diseases including ischemic heart disease (IHD) and peripheral arterial disease (PAD), plasma AM concentration and other inflammatory markers such as high sensitive C-reactive protein (CRP) and interleukin (IL)-6 were examined. The plasma AM level was not altered by the absence or presence of each of four major risk factors, i.e., hypertension, diabetes mellitus, hyperlipidemia, and smoking and its level was not significantly correlated with blood pressure, plasma glucose, or serum lipid levels. The patients with IHD had a significantly higher concentration of plasma AM than those without IHD. The AM level in subjects with PAD was also increased significantly compared with those without PAD. The plasma AM was strongly correlated with inflammatory parameters such as CRP and IL-6. Among AM, CRP, and IL-6, however, only AM was an independent predictor for both IHD and PAD by multiple logistic regression analysis. Our findings suggest the possibility that plasma AM is a novel sensitive marker for the presence of vascular lesions in patients with atherosclerotic risks. PMID- 15350701 TI - Transport of angiotensin peptides across the Caco-2 monolayer. AB - The bidirectional transport of the angiotensin peptides--des-Asp-angiotensin I (DAAI), angiotensins III and IV--were studied using human intestinal Caco-2 monolayers. The peptides had low permeability rates but were relatively stable to enzymatic hydrolysis. DAAI was transported by diffusion while angiotensins III and IV were transported by an energy requiring, carrier-mediated process. The physicochemical properties and solution conformations of the peptides were investigated in an attempt to establish structure-transport correlations. Among the three peptides, DAAI was the most hydrophobic, had the highest hydrogen bonding potential and was the only peptide to have a random solution conformation, as determined from circular dichroism, two-dimensional (1)H NMR and molecular modelling. On the other hand, the more hydrophilic angiotensin IV had less hydrogen bonding potential and a solution conformation characterized by a beta turn. These factors may influence the transport characteristics of DAAI and angiotensin IV. PMID- 15350702 TI - Differential levels of "urotensin-II-like" activity determined by radio-receptor and radioimmuno-assays. AB - Plasma and urinary levels of "urotensin(U)-II-like" substances determined in healthy human volunteers were 12.4 +/- 0.6 ng/ml and 2.2 +/- 0.3 ng/ml by RIA, an order of magnitude lower than that seen by RRA, 167.5 +/- 9.5 ng/ml and 65.2 +/- 4.3 ng/ml. HPLC demonstrated the existence of at least three prominent activity peaks in plasma and urine, the more hydrophobic of which did not co-elute with U II, degradation products or URP. RRA and RIA recognized these peaks with contrasting efficacy. As such, published levels of "U-II-like" activity should be interpreted with caution until a better understanding is obtained regarding what species specific RIA and RRA assay reagents interact with. PMID- 15350703 TI - Study in vitro and in vivo of nociceptin/orphanin FQ(1-13)NH2 analogues substituting N-Me-Gly for Gly2 or Gly3. AB - In the present study, two analogues containing N-Me-Gly (Sarcosine, Sar) were synthesized to further investigate the structural-activity relationships of orphanin FQ/nociceptin (OFQ/NC, NC). The replacement of Gly(2) or Gly(3) with Sar increased the flexibility and decreased the hydrophobicity of the N-terminal tetrapeptide. The activity of the analogues was investigated in a series of assays in vivo and in vitro. [Sar(2)]NC(1-13)NH(2) was found to (1) produce dose dependent inhibition of the electrically induced contraction in MVD assay (pEC(50) = 6.14); (2) produce significant hyperalgesia effects in a dose dependent manner when intracerebroventricularly (i.c.v.) injected in mice. The inhibitive effects of [Sar(2)]NC(1-13)NH(2) in MVD assay could be significantly antagonized by [Nphe(1)]NC(1-13)NH(2), and partially antagonized by naloxone; the hyperalgesic effect of [Sar(2)]NC(1-13)NH(2) could be significantly antagonized by naloxone, and partially antagonized by [Nphe(1)]NC(1-13)NH(2). On the contrary, [Sar(3)]NC(1-13)NH(2) showed no effects in these assays. All the findings suggest that the flexibility of the peptide bond between Phe(1) and Gly(2) and between Gly(2) and Gly(3) play an important role in NC-OP(4) receptor interaction, and the hydrophobicity of the N-terminal tetrapeptide showed no significant effect on this interaction. The present work also helps to provide a novel method to elucidate structural and conformational requirements of the opioid peptide-receptor interaction. PMID- 15350704 TI - Modulation of anxiety-related behaviors by mu- and kappa-opioid receptor agonists depends on the social status of mice. AB - This study was aimed to determine the effects of mu- and kappa-opioid receptor activation in relation to the social status of mice, being a winner with repeated experience of victories or a loser with repeated experience of social defeats. The behaviors of the animals were assessed in a social encounter test measuring the communicative behavior towards a familiar and an unfamiliar partner behind a perforated transparent partition (partition test) and in an elevated plus-maze test estimating the anxiety level of mice. Placebo and graded doses of the mu opioid receptor agonist DAMGO (0.5 and 2 mg/kg s.c.) and the kappa-opioid receptor agonist U-50,488H (0.6, 1.25, and 2.5 mg/kg s.c.) were administered to the control mice, winners and losers in two experiments. In the partition test, the winners spent somewhat more time and the losers less time than the controls in the vicinity of their partner probably related to a lower and higher level of anxiety respectively. In the plus-maze test the losers appeared to have a somewhat higher anxiety level than the controls and winners. In both tests DAMGO produced anxiogenic-like effects in the winners and the controls, but not in the losers. Winners hardly responded to treatment with U-50,488H, while the losers responded dose dependently with an anxiolytic-like effect in both tests. It is concluded that anxiety-like responses in mice are differentially affected by stimulation of mu- and kappa-opioid receptors and that the effects depend on the social status of the animals. PMID- 15350705 TI - A novel ribonuclease from fruiting bodies of the common edible mushroom Pleurotus eryngii. AB - A ribonuclease with a temperature optimum of about 70 degrees C and a pH optimum of 6.5 was isolated from fruiting bodies of the mushroom Pleurotus eryngii. The ribonuclease was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and S-Sepharose. It possessed a molecular mass of 16 kDa, and exhibited higher ribonucleolytic activity toward poly A and poly G and lower ribonucleolytic activity toward poly C and poly U. Its N-terminal sequence was distinctly different from those of other mushroom ribonucleases, and resembled that of Pleurotus tuber-regium only by 40%. Furthermore, its thermostability characteristics, polyhomoribonucleotide specificity and molecular mass were dissimilar to those of other mushroom ribonucleases. PMID- 15350706 TI - Identification of an Adrenomedullin precursor fragment in plasma of sepsis patients. AB - Adrenomedullin and PAMP are potent vasodilatory peptides derived from a common larger precursor peptide. Elevation of circulating levels of both peptides has been described for diseases involving dysfunction of the cardiovascular system. However, the reliable quantification has been hampered by their short half-life times and - as known for Adrenomedullin -- the existence of a binding protein. Here we report the identification of another peptide derived from the Adrenomedullin precursor, termed proADM 45-92, which is present in large concentrations in plasma of septic shock patients. This peptide is produced in stoichiometric amounts to Adrenomedullin and PAMP, but -- contrary to them -- is apparently non-functional and stable. Thus, proADM 45-92 represents a suitable diagnostic target which could be used to assess the concentrations of Adrenomedullin gene products released into the bloodstream. PMID- 15350707 TI - The Lowbury lecture: behaviour in infection control. AB - The majority of healthcare-associated infections result from cross-transmission related to inappropriate patient-care practices. Improving practices frequently implies modifying healthcare workers' behaviour, a key challenge of today's infection control. To improve healthcare workers' compliance with practices, infection control should learn from the behavioural sciences. Social cognitive models can help to improve our understanding of human behaviour. Cognitive determinants that shape behaviour are acquired through the socialization process and are susceptible to change. Some models have been applied to evaluate predictors of health behaviour but, so far, none have been successfully applied to explain behaviour in the field of infection control. Successful strategies to improve infection control practices result from their multidimensional aspect. Similarly, social models that include several levels of cognitive determinants have more chance of success to explain change in behaviour. Concrete examples applied to infection control issues are presented, including special references to hand hygiene behaviour. The theory of ecological perspective, based on the idea that behaviour is viewed as being affected by and affecting multiple levels of influence, and that it both influences and is influenced by the social environment, seems promising to explain behaviour modification. Studies are needed to assess the key determinants of infection control practices and behaviour promotion among the different populations of healthcare workers, and to implement and evaluate the impact of the different components of multimodal programmes to promote optimal infection control practices. PMID- 15350708 TI - Comparison of the effect of closed versus open endotracheal suction systems on the development of ventilator-associated pneumonia. AB - The aim of this study was to compare the effect of closed versus open endotracheal suction systems on the development of ventilator-associated pneumonia (VAP). A prospective, randomized, controlled trial was performed in a medical intensive care unit (MICU) of a university hospital in patients who received mechanical ventilation for more than 48 h. Patients were randomized to receive endotracheal suction with either closed catheters (closed suction group; N-41) or single-use catheters (open suction group; N=37). Cultures were taken from the ventilator tubing of 42 patients to determine the rate of colonization. There was no difference between the groups in terms of the frequency of development of VAP, mortality in the MICU, length of MICU stay and duration of mechanical ventilation. Thirteen patients in the open suction group and 16 patients in the closed suction group became colonized (P=0.14). The colonization rates by Acinetobacter spp. and Pseudomonas aeruginosa were more frequent in the closed suction group than in the open suction group (P<0.01 and P=0.04, respectively). In conclusion, closed endotracheal suction resulted in increased colonization rates of ventilator tubing with multi drug-resistant micro-organisms but did not increase the development of VAP and MICU outcome compared with open endotracheal suction. PMID- 15350709 TI - Staphylococcus aureus nasal carriage in patients on haemodialysis: role of cutaneous colonization. AB - We performed a prospective study of Staphylococcus aureus nasal carriage in patients on chronic haemodialysis to determine the role of cutaneous colonization in the aetiology of recurrent nasal colonization. From February 2000 to September 2001, 71 patients on chronic haemodialysis in the dialysis unit at a university hospital were screened monthly for S. aureus nasal carriage. Carriers received nasal mupirocin for five days and were tested for nasal and cutaneous carriage two days later and monthly thereafter. Using genotyping results, recurrence was defined as relapse if pretreatment and subsequent nasal isolates were clonally identical; if the isolates were different, it was considered recolonization. Thirty-nine patients (55%) were nasal carriers: 11 initially and 28 during follow up. Among the mupirocin-treated patients, the eradication of S. aureus nasal carriage rate was 88.5%. Nasal recurrence was documented in 17 patients (43.5%), and S. aureus nasal strains were available for molecular typing in 14 patients with a total of 23 recurrence episodes. On the basis of pulsed-field gel electrophoresis analysis, 16 (70%) recurrence episodes were considered relapses and seven were considered (30%) recolonizations. Among the episodes of relapse, prior cutaneous colonization was detected in only three cases. In haemodialysis patients, the majority of nasal carriage recurrences after mupirocin therapy were due to relapses. Cutaneous colonization does not appear to be relevant in the development of these relapses. PMID- 15350710 TI - Implications of colonization of vancomycin-resistant enterococci (VRE) in renal dialysis patients. Learning to live with it? AB - Vancomycin-resistant enterococci (VRE) commonly colonize, but less frequently infect, debilitated patients, such as those on chronic renal dialysis. The emergence of VRE amongst our cohort of renal replacement therapy patients posed considerable challenges in our attempts to prevent spread. Although 60 of 451 (13%) patients became colonized, only two patients required systemic antibiotics for confirmed or suspected invasive infection. Mortality and inpatient stay was greater in VRE-positive compared with VRE-negative patients (50% versus 10%) and patients who were screened on three or more occasions were likely to remain positive (e.g. 56% of patients screened on six occasions were positive). The application of recommended guidelines for the control of VRE, however, severely disrupted our renal dialysis programme and therefore had to be abandoned. As patients on renal dialysis are more likely to acquire VRE, remain colonized, require antibiotics and require regular inpatient or outpatient care more frequently than other patients, control measures should be adapted to minimize spread but not disrupt important and essential medical services. PMID- 15350711 TI - Double-blind comparative study on the care of the neonatal umbilical cord using 80% ethanol with or without chlorhexidine. AB - We conducted a double-blind comparative study on the effectiveness of 80% ethanol (EtOH), with or without chlorhexidine (CHD), in the prevention of neonatal umbilical colonization by Staphylococcus aureus, the most frequently isolated pathogen on the neonatal umbilicus in the early period. A total of 100 neonates born at the National Tokyo Medical Centre from March to May 2000 and nursed at a maternity ward were enrolled. Forty-eight were randomly allocated to the group for whom umbilical cord disinfection was performed using 80% EtOH containing 0.5% CHD (CHD group) and 52 to disinfection with 80% EtOH alone (EtOH group). The mothers of the neonates and the nursing staff were unaware as to which disinfectant was being used. Disinfection of the umbilicus and the surrounding area was done immediately after birth and twice daily thereafter, after bathing and in the evening, throughout the hospitalization period. Specimens for bacterial culture were taken from the umbilical cord and surrounding skin on day 4 or 5 after birth. As a disinfectant susceptibility test, we checked the minimum killing concentration (MKC) of CHD and EtOH. There was no statistically significant difference between the two groups with respect to sex, gestation period, birthweight, APGAR score or delivery method. In the CHD group, S. aureus was isolated from 25% of the patients, while it was isolated from 57.7% in the EtOH group (P<0.001). In the CHD group, 50% of the S. aureus strains were MRSA, compared with 73.3% in the EtOH group (non-significant). All the S. aureus strains were killed by the combination of both CHD and EtOH at the concentrations used. In terms of the MKC90, there was no significant difference between the CHD group and the EtOH group. For the daily care of the neonatal umbilicus, disinfection using 80% EtOH containing CHD was found to be more effective than that using 80% EtOH alone in preventing colonization by S. aureus. PMID- 15350712 TI - Use of palivizumab to control an outbreak of syncytial respiratory virus in a neonatal intensive care unit. AB - To evaluate the safety and effectiveness of a humanized respiratory syncytial virus (RSV) monoclonal antibody (palivizumab) to control an outbreak of RSV in a neonatal intensive care unit (NICU), we retrospectively analysed two RSV outbreaks. Between 11 November 1998 and 18 March 1999, two separate RSV outbreaks occurred in a large (26 beds) NICU. All procedures for preventing nosocomial spread of RSV (including the use of palivizumab in the second outbreak) were retrospectively analysed. The cumulative incidence (CI), secondary attack rate (SAR) and risk ratio of infection were determined before and after the use of palivizumab for all patients and for those with gestational age below and above 32 weeks in the NICU during the second outbreak. Standard infection control measures were effective in the first outbreak (three cases). In the second outbreak, after three index cases, five additional infants were newly RSV infected within one month. Three infants had RSV pneumonia and required mechanical ventilation; one infant died. Standard infection control procedures were initiated from the beginning of this outbreak. Palivizumab was given to all infants in the NICU after the fifth case was identified. CI was 2.4% in the first 15 days and 10.5% in the second, and SAR was 2.9 per thousand in the first 15 days and 14.1 per thousand in the second, both dropping to zero after the administration of palivizumab. The risk ratio of infection was 4.65 times higher in infants under 32 weeks gestational age. After the use of palivizumab, there were no additional identified cases. In addition to careful infection control procedures, the use of palivizumab might have contributed to arresting the outbreak of RSV infection in the NICU, suggesting that it could be an additional resource in the control of severe nosocomial RSV outbreaks. PMID- 15350713 TI - Effects of cleaning and disinfection in reducing the spread of Norovirus contamination via environmental surfaces. AB - A reverse transcriptase polymerase chain reaction assay was used to study the transfer of Norovirus (NV) from contaminated faecal material via fingers and cloths to other hand-contact surfaces. The results showed that, where fingers come into contact with virus-contaminated material, NV is consistently transferred via the fingers to melamine surfaces and from there to other typical hand-contact surfaces, such as taps, door handles and telephone receivers. It was found that contaminated fingers could sequentially transfer virus to up to seven clean surfaces. The effectiveness of detergent- and disinfectant-based cleaning regimes typical of those that might be used to decontaminate faecally contaminated surfaces and reduce spread of NV was also compared. It was found that detergent-based cleaning with a cloth to produce a visibly clean surface consistently failed to eliminate NV contamination. Where there was faecal soiling, although a combined hypochlorite/detergent formulation at 5000 ppm of available chlorine produced a significant risk reduction, NV contamination could still be detected on up to 28% of surfaces. In order consistently to achieve good hygiene, it was necessary to wipe the surface clean using a cloth soaked in detergent before applying the combined hypochlorite/detergent. When detergent cleaning alone or combined hypochlorite/detergent treatment failed to eliminate NV contamination from the surface and the cleaning cloth was then used to wipe another surface, the virus was transferred to that surface and to the hands of the person handling the cloth. In contrast, were surfaces where contaminated with NV-infected faecal suspension diluted to 1 in 10 and 1 in 80, intended to simulate surfaces that have become contaminated after secondary transfer, treatment with a combined bleach/detergent formulation, without prior cleaning, was sufficient to decontaminate surfaces and prevent transfer. PMID- 15350714 TI - Manual versus automated methods for cleaning reusable accessory devices used for minimally invasive surgical procedures. AB - We undertook a simulated-use study using quantitative methods to evaluate the cleaning efficacy of ported and non-ported accessory devices used in minimally invasive surgery. We chose laparoscopic scissors and forceps to represent worst case devices which were inoculated with artificial test soil containing 10(6) cfu/mL Enterococcus faecalis and Geobacillus stearothermophilus and allowed to dry for 1 h. Cleaning was performed manually, as well as by the automated SI-Auto Narrow lumen cleaner. Manual cleaning left two- to 50-fold more soil residuals (protein, haemoglobin and carbohydrate) inside the lumen of non-ported versus ported laparoscopic accessory devices. The SI-Auto Narrow lumen cleaner was more efficient than manual cleaning and achieved >99% reduction in soil parameters in both non-ported (using retro-flushing) and ported laparoscopic devices. Only the automated cleaning of ported devices achieved 10(3)-10(4)-fold reduction in bacterial numbers. Sonication alone (no flushing of inner channel) did not effectively remove soil or organisms from the inner channel. Our findings indicate that non-ported accessory devices cannot be as reliably cleaned as ported devices regardless of the cleaning method used. If non-ported accessory devices are reprocessed, they should be cleaned using retro-flushing in an automated narrow lumen cleaner. PMID- 15350715 TI - A survey on the pathogenic fungi in soil samples of potted plants from Sari hospitals, Iran. AB - A total of 23 soil samples of potted plants was collected from hospitals in Sari, Iran. Each sample contained approximately 200 g soil, taken from a depth of 0-10 cm of the pots. Samples were analysed by two different methods. (1) Culture on Sabouraud's dextrose agar medium containing chloramphenicol (SC); cultured fungi were identified by macroscopic and microscopic characterization. (2) The hair baiting technique (HBT) was used to isolate the keratinophilic fungi. After culture on SC medium, 100% of plates were positive for fungal growth, and in total 1150 colonies with 13 different types of fungi were isolated. Penicillium spp. (52%), Acremonium spp. (20%), Paecilomyces spp. (11.9%), Cladsporium spp. (3.7%) and Aspergillus spp. (3.1%) were the predominant fungal species. Rhizopus spp. (0.1%) were less frequent. Keratinophilic fungi such as Microsporum gypseum, Microsporum cookei and Chrysosporium spp. were isolated using HBT. The presence of pathogenic fungi such as Cladosporium spp, Aspergillus spp, M. gypseum, and M. cookei in potted plants in hospitals represents a potential source of nosocomial infection. PMID- 15350716 TI - Nosocomial and community-acquired Enterobacter cloacae bloodstream infection: risk factors for and prevalence of SHV-12 in multiresistant isolates in a medical centre. AB - In a medical centre in northern Taiwan, 60 patients had bloodstream infection caused by Enterobacter cloacae from 1 January 2002 to 30 April 2003. Forty (66.7%) were nosocomial and 26 were caused by multiresistant isolates. Twenty patients died due to the infection. Central venous catheterization and mechanical ventilation were relative risks for nosocomial E. cloacae infection. Age and mechanical ventilation were risk factors for multiresistant E. cloacae infection. Mortality was associated with multiresistant isolates and polymicrobial infection. Pulsed-field gel electrophoresis (PFGE) analysis showed, the 26 multiresistant isolates comprised 12 different types, with type A predominating (12 isolates). Excluding the patients infected with PFGE type A, central venous catheterization was a relative risk for infection, and polymicrobial infection was a risk factor for mortality. All but one of the 26 multiresistant isolates had the extended-spectrum beta-lactamase SHV-12. TEM-1 and ampC beta-lactamase genes were also detected in 25 of the 26 multiresistant isolates. Southern blotting indicated that the SHV-12 gene was located on plasmids. Eleven of the 26 multiresistant isolates had minimum inhibitory concentrations (MIC) > or =16 mg/L for cefepime, which was reduced by the addition of sulbactam for most isolates, resulting in susceptibility. The combination of cefepime and sulbactam may be effective in the treatment of multiresistant E. cloacae bloodstream infection. PMID- 15350717 TI - Alcoholic fixation of blood to surgical instruments-a possible factor in the surgical transmission of CJD? AB - While developing a new protein removal test for the quality control of surgical instrument cleaning, it was noted that alcohol firmly binds blood to stainless steel. Creutzfeldt-Jacob disease is one of the transmissible spongiform encephalopathies (TSE) that has been transmitted between humans and chimpanzees by electroencephalogram electrodes, previously 'sterilized' using ethanol and formaldehyde. Although ethanol has a bactericidal action, it also binds protein to metal. Prion proteins found in TSE are thought to be the causal agents of spongiform disease and it is likely that these proteins are also bound to the stainless steel of surgical instruments by alcohols. Where spongiform disease is a possibility, alcohol, and probably formaldehyde, should not be used to decontaminate neurosurgical instruments. PMID- 15350718 TI - Prevention of percutaneous endoscopic gastrostomy site infections caused by methicillin-resistant Staphylococcus aureus. AB - Percutaneous endoscopic gastrostomy (PEG) is widely used to maintain enteral nutrition in patients who are unable to swallow. Peristomal wound infection is the most common complication of this procedure. In a hospital endemic for methicillin-resistant Staphylococcus aureus (MRSA), MRSA can be the most common organism associated with these infections. We have evaluated a strategy consisting of screening, skin decontamination and glycopeptide prophylaxis for preventing PEG-site infections. None of the 34 patients who received the decontamination protocol and glycopeptide prophylaxis (Group A) developed PEG site infections within one month of surveillance. Two patients were infected with MRSA after that period. One of seven patients who received the decontamination protocol alone (Group B) was infected within the period of surveillance, while another patient was infected after that period. Both were infected with MRSA. None of nine patients who received glycopeptide prophylaxis alone (Group C) were infected. The results suggest that the strategy of screening, decontamination and glycopeptide prophylaxis is effective in the prevention of PEG-site infections with MRSA. Further trials are necessary to confirm these findings. PMID- 15350719 TI - Does intrapartum antibiotic prophylaxis decrease the incidence of maternal group B streptococcal infections? PMID- 15350720 TI - The value of using chlorhexidine soap in a controlled trial to eradicate MRSA in colonized patients. PMID- 15350721 TI - Comparison of antibiotic dispensing data with administration data in intensive care. PMID- 15350722 TI - Barrel suction filters are not a source of intra-operative infection in total hip replacement. PMID- 15350723 TI - Decrease in Staphylococcus aureus surgical-site infection rates after orthopaedic surgery after intranasal mupirocin ointment. PMID- 15350724 TI - Blood pressure cuffs as potential reservoirs of extended-spectrum beta-lactamase VEB-1-producing isolates of Acinetobacter baumannii. PMID- 15350725 TI - Immunoprophylaxis or acyclovir prophylaxis against hospital-acquired varicella. PMID- 15350726 TI - Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays. AB - A single tube, fluorogenic probe-based, real-time quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) assay was developed for detection and quantitation of bovine respiratory syncytial virus (BRSV) using BioRad's iCycler iQ. Real-time Q-RT-PCR was compared with quantitative competitive RT-PCR (QC-RT-PCR) and viral titers. Viral mRNA levels were measured in BRSV-infected bovine turbinate cell lysate harvested at eight time points (1.5, 6, 12, 24, 36, 48, 60, 72 h) post-infection. A homologous BRSV cRNA standard was used for quantitation of the mRNA by plotting a standard curve of cycle threshold (Ct) values versus standard 10-fold dilutions of cRNA of known concentrations. Detection as low as 171 copies/microl of standard BRSV cRNA was possible. For QC-RT-PCR, a competitor RNA molecule having a deletion was designed and used for quantitation of the BRSV viral mRNA. The results of real-time Q-RT PCR and QC-RT-PCR assays showed a positive correlation. Real-time Q-RT-PCR was a sensitive, specific, rapid, and efficient method that eliminates the post-PCR processing steps when compared to QC-RT-PCR. Quantitation of BRSV using real-time Q-RT-PCR will have application in studies aimed at understanding the pathogenesis of BRSV. PMID- 15350727 TI - Evaluation of different assays for the detection of parvovirus B19 DNA in human plasma. AB - Human parvovirus B19 is a frequent contaminant of blood and plasma-derived medicinal products and transmission of this virus has been shown to occur through the administration of contaminated products. Inactivation of the virus has proved difficult and as a consequence, manufacturers of blood products have implemented screening measures to reduce the load of parvovirus B19 in manufacturing plasma pools by the use of nucleic acid amplification techniques (NAT). We present data comparing the performance of two commercially available kits for the detection and quantitation of parvovirus B19 DNA using the LightCycler, and determine their applicability for the detection of recently discovered variants of the virus. Parvovirus B19 DNA was readily quantified using both commercial assays. However, one kit failed to detect any of the variant viruses. The second kit detected the variant viruses although there was a marked difference in the sensitivity of detection of the different virus genotypes. To improve the detection of these variant viruses a novel assay has been developed and data are presented to show its use for screening pooled plasma for the presence of parvovirus B19 DNA and the variants identified recently. PMID- 15350728 TI - Development and use of an internal standard for oyster herpesvirus 1 detection by PCR. AB - Oyster samples were examined using a competitive PCR method in order to detect and quantify oyster herpesvirus 1 (OsHV-1) DNA. Quantitation of viral DNA by competitive PCR assay was based on co-amplification of OsHV-1 DNA and a competitor where a known amount of competitor DNA was present in the same reaction mixture. The competitor was engineered so that it differs in length (deletion of 76 base pairs) from the viral DNA. The assay allowed the detection of 1 fg of viral DNA among 0.5 mg of oyster tissues. The method was used to demonstrate the absence of PCR inhibitors in oyster spat ground tissues. PCR inhibition was observed in adult oyster samples when the same tissue preparation procedure was used. On the contrary, classical phenol/chloroform DNA extraction from adult oyster tissues allowed co-amplification of the internal standard competitor and the viral DNA. The method was successfully used to demonstrate the presence of viral DNA in asymptomatic adult oysters. Quantitation of OsHV-1 DNA in infected spat and asymptomatic adult oysters was also carried out. Viral DNA (1.5-325 pg) were detectable in 0.5 mg of oyster tissues in adults. The amounts of viral DNA in infected oyster spat varied from 750 pg to 35 ng per 0.5 mg of ground tissues. PMID- 15350729 TI - Development of a one tube-one step RT-PCR protocol for the detection of seven viroids in four genera: Apscaviroid, Hostuviroid, Pelamoviroid and Pospiviroid. AB - A one tube-one step RT-PCR was developed for the detection of seven viroids (Apple scar skin viroid, Apple dimple fruit viroid, Pear blister canker viroid, Hop stunt viroid, Chrysanthemum stunt viroid, Citrus exocortis viroid and Peach latent mosaic viroid) in four genera that infect eight plant species. The efficiency and specificity of this method were optimized by the use of Moloney murine leukemia virus (M-MLV) reverse-transcriptase and HotStarTaq DNA polymerase which allowed increase sensitivity of viroid detection. The method was assessed with 56 viroid-infected field plants. The multiplex one tube-one step RT-PCR has the advantage of requiring less hands-on time to set up an assay than standard multiplex one, it also reduces the possibility of false positive tests because all steps are performed in the same tube thus, avoiding cross-contamination. The method may be used routinely for viroid detection in sanitary and certification programmes. PMID- 15350730 TI - Immunodiagnosis of Prune dwarf virus using antiserum produced to its recombinant coat protein. AB - Certification represents the first line of defense against fruit tree viruses. For certification or surveys dealing with large number of samples, ELISA is still considered the technique of choice and requires a continuous supply of good quality antibodies. Prune dwarf virus (PDV) is among the major viruses affecting stone fruits; it belongs to the genus Ilarvirus named so for its isometric labile particles. Recombinant DNA technology was investigated for production of PDV antiserum to avoid labile virus purification and virus maintenance problems. The PDV coat protein gene (CP) was cloned into a protein expression bacterial plasmid vector which allowed a good level of expression of up to 2mg native protein/L culture. The recombinant PDV CP was injected into rabbits and the crude antiserum was successfully used in indirect ELISA at dilutions of up to 1:5000 to detect PDV in infected leaf samples. Similar results were obtained in dot blot immunoassays (DBIA). The antibodies were used in double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) and results were comparable to a reference commercial kit. The crude antiserum was efficiently used for coating ELISA plates, thereby reducing test costs. PMID- 15350731 TI - Development of homologous viral internal controls for use in RT-PCR assays of waterborne enteric viruses. AB - Enteric viruses often contaminate water sources causing frequent outbreaks of gastroenteritis. Reverse transcription-polymerase chain reaction (RT-PCR) assays are commonly used for detection of human enteric viruses in environmental and drinking water samples. RT-PCR provides a means to rapidly detect low levels of these viruses, but it is sensitive to inhibitors that are present in water samples. Inhibitors of RT-PCR are concentrated along with viruses during sample processing. While procedures have been developed to remove inhibitors, none of them completely remove all inhibitors from all types of water matrices. This problem requires that adequate controls be used to distinguish true from potentially false-negative results. To address this problem, we have developed homologous viral internal controls for hepatitis A virus (HAV), poliovirus, Norwalk virus and rotavirus. These internal controls can be used in RT-PCR assays for the detection of the above viruses by competitive amplification, thereby allowing the detection of false negatives in processed water samples. The internal controls developed in this study were successfully tested with virus seeded environmental water sample concentrates. PMID- 15350732 TI - Development of immunocapture reverse transcription loop-mediated isothermal amplification for the detection of tomato spotted wilt virus from chrysanthemum. AB - An immunocapture reverse transcription loop-mediated isothermal amplification (IC/RT-LAMP) was developed for the detection of tomato spotted wilt virus (TSWV) from chrysanthemum. This method enabled sensitive, reproducible and specific detection of TSWV from chrysanthemum plants. In the RT-LAMP method, TSWV genomic RNA could be amplified under isothermal (65 degrees C) conditions within 1 h. The resulting amplicons were detected by the measurement or observation of the turbidity of the reaction mixture without gel electrophoresis. IC/RT-LAMP was 100 times more sensitive than IC/RT-PCR. PMID- 15350733 TI - A universal microarray for detection of SARS coronavirus. AB - Severe acute respiratory syndrome (SARS) is caused by the SARS coronavirus (SARS CoV). There are many point mutations among SARS-CoV genome sequences. Previous studies suggested that the mutations are correlated closely with the SARS epidemic. It was found that the bases of six nucleotide positions (nt9404, nt9479, nt19838, nt21721, nt22222 and nt27827) with high-mutation rate have an important relationship with the SARS epidemic. For viral detection as well as genotyping, a universal microarray system was developed that combines RT-PCR and ligase detection reaction (LDR). The Zip Codes attached covalently to a slide remain constant and their complementary Zip Codes (cZip Codes) can be used for tagging target sequence, making the microarrays universal. The discriminating oligonucleotides contain on the 5' end "cZip Codes" that are used to direct LDR product to specific Zip Codes attached covalently to a slide. Since Zip Codes have no homology to either the target sequence or to other sequences in the genomes of both human host and SARS-CoV, there was no false signal due to mismatch hybridizations. 20 samples assayed with the universal microarray were confirmed by DNA sequencing, demonstrating that this microarray system is a promising diagnostic tool for detection and genotyping of the SARS-CoV. PMID- 15350734 TI - Highly reproducible transient transfections for the study of hepatitis B virus replication based on an internal GFP reporter system. AB - High throughput studies of hepatitis B virus (HBV) replication are limited by the absence of convenient and reliable in vitro systems that can be utilised to examine mutations emerging as a consequence of selective pressures such as drug therapy, immune response and genotypic evolution. We have developed an efficient and reproducible method of transfecting a human hepatoma cell line (Huh7) with a replication competent clone of HBV, which utilises an internal reporter of transfection efficiency. Wild type (WT) and a common lamivudine (LMV) resistant mutant virus characterised by a methionine to isoleucine mutation in the highly conserved YMDD motif of the polymerase gene were used to highlight the utility of this in vitro transfection procedure in studying common HBV mutations. The presence of an internal GFP reporter increased the efficiency of generating highly reproducible results and facilitated multiple forms of analysis from a single transfection accurately. PMID- 15350735 TI - Development of recombinant capsid antigen/transmembrane epitope fusion proteins for serological diagnosis of animal lentivirus infections. AB - Among animal lentiviruses, Feline immunodeficiency virus (FIV), Equine infectious anaemia virus (EIAV) and Small ruminant lentiviruses (SRLV) are important pathogens associated with a variety of clinical pictures including immunodeficiency, anaemia, arthritis, pneumonia. The detection of viral antibody response represents a practical diagnostic approach in all lentivirus infections since they remain detectable long life. Capsid antigen (CA) is the major viral core protein and specific antibodies against this antigen are usually first recognised in infected sheep, goat and horse, remaining detectable for long period. Transmembrane (TM) domain of envelope glycoprotein contains a well conserved motif known to form an immunodominant epitope in several lentiviruses. In this study a simple strategy was developed to express the entire CA and the TM epitope in a single fusion protein from equine, feline and small ruminant lentiviruses in prokaryotic system and evaluated the diagnostic utility of a purified preparation in an indirect ELISA for each of the three infections. Results demonstrate that, for FIV and SRLV infections, the combination of CA and TM fractions increases the sensitivity of diagnostic tests based only on CA. The corresponding CA/TM antigen from EIAV showed excellent agreement with Coggins test. PMID- 15350736 TI - Analytical performance of and real sample analysis with an HBV gene visual detection chip. AB - A novel hepatitis B virus (HBV) gene detection chip has been developed. The HBV specific probes immobilized on glass slides were hybridized with polymerase chain reaction (PCR) products of different serum samples. The hybridization signal can be easily visualized upon a sandwich assay with nanoparticle amplification. The analytical performance (e.g., specificity, sensitivity, and accuracy) of this method has been evaluated. The chip-based detection method possesses a greater sensitivity and a better reproducibility than some of the conventional immunological or molecular biological methods (e.g., enzyme-linked immunosorbent assay, ELISA) and is simple, cost-effective, and highly selective. PMID- 15350737 TI - Inactivation of the coronavirus that induces severe acute respiratory syndrome, SARS-CoV. AB - Severe acute respiratory syndrome (SARS) is a life-threatening disease caused by a novel coronavirus termed SARS-CoV. Due to the severity of this disease, the World Health Organization (WHO) recommends that manipulation of active viral cultures of SARS-CoV be performed in containment laboratories at biosafety level 3 (BSL3). The virus was inactivated by ultraviolet light (UV) at 254 nm, heat treatment of 65 degrees C or greater, alkaline (pH > 12) or acidic (pH < 3) conditions, formalin and glutaraldehyde treatments. We describe the kinetics of these efficient viral inactivation methods, which will allow research with SARS CoV containing materials, that are rendered non-infectious, to be conducted at reduced safety levels. PMID- 15350738 TI - Comparison of three current viral load assays for the quantitation of human immunodeficiency virus type 1 RNA in plasma. AB - The LCx HIV RNA quantitative assay (Abbott Laboratories, Delkenheim, Germany) was compared with the Versant HIV-1 RNA 3.0 (bDNA) assay (Bayer, Tarrytown, NY) and the COBAS Amplicor HIV-1 Monitor v1.5 test (Roche Diagnostics, Branchburg, NJ), using plasma samples of various viral load levels from HIV-1-infected patients. Considering the lower limit of the linear range of 50 copies/ml of both assays, the detection range of the LCx was 127/151 (84.1%) versus the 131/151 (86.8%) of the bDNA 3.0 assay, while overall agreement between the two assays was 93.4% (141/151). LCx and bDNA 3.0 results were found to be strongly correlated (r = 0.96). The fitted regression line was described by the equation log10(LCx copies/ml) = 0.05 + 1.06 x log10(bDNA 3.0 copies/ml) with 95% CI for the estimated slope and intercept at 1.01, 1.12 and -0.16, 0.26, respectively. Similarly, the detection range of the LCx was 115/148 (77.7%) versus the 128/148 (86.5%) of the Monitor v1.5 test. A 91.2% concordance (135/148) was observed between these two assays at a cut-off of 50 copies/ml. LCx and Monitor v1.5 results were highly correlated (r = 0.96). The fitted regression line was described by the equation log10(LCx copies/ml) = 0.06 + 1.03 x log(10)(Monitor v1.5 copies/ml) with 95% CI for the estimated slope and intercept at 0.97, 1.09 and -0.16, 0.28, respectively. PMID- 15350739 TI - Detection of plant viruses using a surface plasmon resonance via complexing with specific antibodies. AB - The use of instrumental systems based on the surface plasmon resonance (SPR) for rapid diagnosis of intact plant viruses (in particular, tobacco mosaic virus (TMV)) is considered. A new approach using detection of viral antigen and antibody (IgG) complexes formed during the preincubation step (instead of their consecutive application in classical approach) is discussed. A comparison between signal level registered from the mixture of virus and specific serum and that from the sample without virus (samples deposited onto the sensor surface treated with thiocyanate and protein A Staphylococcus aureus) allows unambiguous detection of viral particles in the material studied. The performance capabilities of the method are discussed and illustrated by quantitative detection of virus in the actual samples (cells homogenate) at high concentration. PMID- 15350740 TI - The use of differential display to isolate viral genomic sequence for rapid development of PCR-based detection methods. A test case using Taura syndrome virus. AB - The purpose of this study was to explore the efficacy of using differential display (DD) to isolate viral genomic sequence using tissues from infected organisms so that a PCR procedure to detect the pathogen may be developed rapidly. The model virus used was the Taura syndrome virus (TSV), a ssRNA virus that cause high rates of mortality at shrimp farms. Two random primers in combination with four anchored primers were used to isolate five cDNAs, ranging in size from 241 to 822 bp, that were differentially expressed in TSV-infected shrimp (Litopenaeus vannamei). PCR experiments revealed that four of the five encoded shrimp genes while the fifth was likely to be a TSV gene. Evidence that the putative TSV sequence is part of the TSV genome was obtained by the 97% sequence identity it shared with the published TSV genome. PCR primers were designed successfully using the differential display sequence to develop a RT-PCR based method to detect TSV. Because differential display does not require physical isolation of the virus and only a small amount of infected sample is needed, the technique may be useful as a method to isolate nucleic acid sequences from emerging pathogens so that PCR primers for their detection may be developed rapidly. PMID- 15350741 TI - Correlation between rules-based interpretation and virtual phenotype interpretation of HIV-1 genotypes for predicting drug resistance in HIV-infected individuals. AB - Drug resistance testing provides useful information for managing HIV-infected patients. Phenotyping could add complementary information to genotyping and occasionally be more useful, although is less available to clinicians. Large paired geno-pheno databases have allowed the prediction of phenotypes from genotypes. However, the accuracy of these virtual phenotypes (vPT) in a clinical setting has not been well assessed yet. We analyzed the concordance between vPT and interpreted genotype (GT) in 105 samples belonging to treatment-experienced HIV-infected patients. A high concordance was seen when examining both non nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI) (r = 0.95 either), while it was lower for nucleoside analogs (r = 0.79). The drugs with lower concordance were abacavir (71.1%), tenofovir (71.5%) and didanosine (71.9%). In 20% of specimens (21/105), the vPT did not provide results for all approved drugs. These were mainly samples with a high number of drug resistance mutations or rare genotypes, which seem to be underepresented in the VircoNET database. Overall, there is good correlation between vPT/GT, especially for PI and NNRTI. The inclusion of additional sequences in the VircoNET database, mainly those derived from heavily treatment-experienced patients and/or from patients failing the most recently approved drugs might improve its performance. PMID- 15350742 TI - Specific detection of lettuce mosaic virus isolates belonging to the "Most" type. AB - Lettuce mosaic virus (LMV)-Most isolates can infect and are seed-borne in cultivars containing the mo1 gene. A reverse transcription and polymerase chain reaction (RT-PCR)-based test was developed for the specific detection of LMV-Most isolates. Based on the complete genome sequences of three LMV isolates belonging respectively to the Most type, the Common type and neither of these two types, three different assays were compared: (i) presence of a diagnostic restriction site in the region of the genome encoding the variable N-terminus of the capsid protein, in the 3' end of the genome, (ii) RT-PCR using primers designed to amplify a cDNA corresponding to a portion of the P1 coding region, in the 5' end of the genome and (iii) RT-PCR using primers designed to amplify a central region of the genome. The assays were performed against a collection of 21 isolates from different geographical origins and representing the molecular variability of LMV. RT-PCR of the central region of the genome was preferred because its results are expected to be less affected by natural recombination between LMV isolates, and it allows sensitive detection of LMV-Most in situations of single as well as mixed contamination. PMID- 15350743 TI - Comparison of TGF-beta 1 concentrations in bronchoalveolar fluid of horses affected with heaves and of normal controls. AB - Airway remodeling may play an important role in heaves pathophysiology. Transforming growth factor-beta 1 (TGF-beta1) is a potent profibrotic cytokine, which might contribute to airway wall thickening and fibrosis of bronchiolar and alveolar submucosa. An ELISA designed for the measurement of human TGF-beta1 was used to measured total TGF-beta1 released in bronchoalveolar lavage fluid (BALF) of normal horses and of those affected with heaves in remission. The specificity of the assay for TGF-beta1 of the horse was confirmed using recombinant equine TGF-beta1. The influence of hay exposure on TGF-beta1 release in the airways was also examined by stabling horses in a dusty environment. TGF-beta1 was found in the BALF of all horses. However, no significant difference between basal concentration of TGF-beta1 in BALF of control horses versus that of horses affected with heaves was found. Furthermore, no differences were identified in these populations 1 and 9 days after allergen challenge. In conclusion, these data indicate that TGF-beta1 is released in BALF fluid of horses in biologically active concentrations. Other studies are necessary for a better definition of the role of this cytokine within the lung, as our study does not establish a causal relationship between TGF-beta1 and the pathophysiology of heaves in the horse. PMID- 15350744 TI - Immunophenotyping of the cutaneous cellular infiltrate after atopy patch testing in cats with atopic dermatitis. AB - Cats with spontaneously occurring atopic dermatitis have clinical and immunocytochemical characteristics compatible with these in humans with atopic dermatitis (AD). The atopy patch test (APT) has proven to be a valuable tool in elucidating the disease process in humans. Additionally, the APT is very specific and bypasses the problem of conflicting results due to differences in chronicity of lesions of AD patients. We adapted the APT for use in cats to explore the suitability of the APT as a tool to study the onset of allergic inflammation in cats with atopic dermatitis. APT were performed in AD cats (n = 6) and healthy cats (n = 10). All cats were patch tested with two allergens in three different dilutions and a diluent control. The allergens for the APT were selected from positive intradermal test and /or prick test results and consisted of: Dermatophagoides farinae, D. pteronyssinus, Tyrophagus putrescentiae, and a grass pollen mixture. APT were read after 10, 24 and 48 h, and punch biopsies for immunohistochemical evaluation were collected at these time points. Macroscopically positive APT reactions were observed in three out of six cats at 24 and/or 48 h with allergen concentrations of 25,000 and 100,000 NU/ml. Reactions were not observed at negative control sites and neither in control animals. A significantly increased number of IL-4+, CD4+, CD3+, MHC class II+ and CD1a+ cells was found in one AD cat with positive APT reactions. Five out of six AD cats had significantly increased IL-4+ T cell numbers at 24 and/or 48 h. Our data indicate that in cats, macroscopically positive patch test reactions can be induced, which have a cellular infiltrate similar to that in lesional skin. We found a high specificity and a macroscopically positive APT reaction in half of the cats, which is similar to what is seen in humans. Hence, the APT in cats might be a useful tool in studying the immunopathogenesis of feline atopic dermatitis. PMID- 15350745 TI - Assessment of anti-bovine IL4 and IFN gamma antibodies to label IL4 and IFN gamma in lymphocytes of the koala and brushtail possum. AB - We assess anti-bovine IL4 and IFN gamma (IFNg) antibodies for their ability to label IL4 and IFNg in koala (Phascolarctos cinereus), common brushtail possum (Trichosurus vulpecula) and mountain brushtail possum (Trichosurus caninus) lymphocytes using flow cytometry and immunohistochemistry to determine their applicability to studies of host response to intracellular pathogens. Anti-IFNg labelled a product of PMA-ionomycin stimulated sheep, koala and possum lymphocytes. High intensity labelling was not reduced by blocking non-specific binding with 10% FCS; and non-permeabilised koala lymphocytes labelled less, demonstrating that the labelled product was intracellular. The anti-IL4 antibody labelled variably more cells than the irrelevant antibody in some stimulated and non-stimulated preparations in all species but intensity of this labelling was similar to that of cells labelled with the irrelevant antibody. In this study, the antibodies did not label frozen or formalin-fixed tissues in a range of immunohistochemical techniques. We expect the anti-IFNg antibody to be effective in evaluating Th1 responses of koalas and possums exposed to various host, pathogen and environmental factors and add to the limited tools available for investigating the pathogenesis of marsupial diseases, especially those caused by intracellular organisms, such as tuberculosis of brushtail possums and chlamydial disease of koalas. PMID- 15350746 TI - Molecular cloning of the viperin gene and its promoter region from the mandarin fish Siniperca chuatsi. AB - A viperin gene has been cloned from the mandarin fish (Siniperca chuatsi). From the first transcription initiation site, the mandarin fish viperin gene extends 3163 nucleotides to the end of the 3' untranslated region, and it contains six exons and five introns. The open reading frame of the viperin transcript has 1062 nucleotides which encode a 354 amino acid peptide. The amino acid sequence of mandarin fish viperin shows high identities with its homologues in teleosts and mammals except for the first 70 amino acids. A characteristic feature in the viperin promoter region was the presence of five putative ICSBP (IRF8) binding sites and one IRF1 binding site. The viperin gene expressed mainly in lymphoid tissues before stimulation, but its expression can be examined in almost all the organs investigated after stimulation with virus or Poly I:C. The expression pattern and promoter sequence may be considered as the indirect evidence that the transcription of viperin is regulated by interferons or interferon induced genes. PMID- 15350747 TI - Evaluation of elutriation and magnetic microbead purification of canine monocytes. AB - An elutriation technique was developed to obtain large quantities of pure canine monocytes. Firstly, peripheral blood mononuclear cells (PBMC) were isolated from whole blood by Ficoll gradient. Then, the PBMC were separated by an elutriation procedure. We demonstrated that these techniques allow the isolation of canine peripheral blood monocytes with a purity of 64% +/- 7.9 when labelled with anti CD14 antibody. This purity increased to 83% +/- 2.2 after separation by magnetic anti-CD14 microbeads. The cell viability was more than 95% and apoptotic cells were less than 10%. The monocytes purified by these methods were functionally active in a mixed leukocyte reaction (MLR). A lymphocyte fraction was obtained directly only by elutriation with an average of 79.9% +/- 10.7 of CD5+, 7.9% +/- 3.5 of CD21+ and 1.78% +/- 2.53 of CD14+. Our results indicate that this elutriation procedure is a safe method to purify monocytes as well as lymphocytes, useful in MLR. PMID- 15350748 TI - Immunophenotypic and functional effects of bunker C fuel oil on the immune system of American mink (Mustela vison). AB - The relationship between exposure to environmental contaminants and immunotoxicity in vulnerable marine species is unknown. In this study, we used American mink (Mustela vision) as a surrogate species for the sea otter to examine the immunotoxic effects of chronic exposure to a low concentration of bunker C fuel oil (500 ppm admixed in the feed for 113-118 days). The mink immune system was monitored over time by flow cytometric analysis for alterations in the immunophenotype of blood lymphocytes and monocytes and by mitogen-stimulated proliferation assays for changes in peripheral blood mononuclear cell function. Fuel oil exposure caused a mild, yet significant (P < 0.05) increase in the absolute numbers of specific peripheral blood lymphocyte subsets (CD3+T cells) and monocytes, an increase in the level of expression of functionally significant cell surface proteins (MHC II, CD18), and an increase in mitogen-induced mononuclear cell proliferative responses. This heightened state of cellular activation along with the increase in specific cell surface protein expression on both the innate and adaptive immune cells is similar to the pro-inflammatory or "adjuvant-like" effect described in laboratory models of polycyclic aromatic hydrocarbon exposure in other species. These results show the benefits of using a controlled laboratory model for detecting and characterizing subtle petroleum oil induced perturbations in immune responses. In addition this study establishes a framework for studying the effects of environmental petroleum oil exposure on the immune system of free-ranging marine mammals. Expansion of these studies to address biolgical significance is warranted. PMID- 15350749 TI - Cryopreserved bovine mammary cells to model epithelial response to infection. AB - Mammary gland epithelial cells are likely to be important effectors in defending against mastitis, yet little is known about their response mechanisms. Here, we describe a cryopreserved bovine mammary epithelial cell model to study the infection response. Primary cell cultures from four Holstein cows were prepared, and frozen after two passages. The cell cultures from each cow were then thawed and maintained separately, yet simultaneously, and exposed to treatments that included infection with Staphylococcus aureus or exposure to LPS from Escherichia coli. A clear inflammatory response was shown by a significant (P < 0.05), dose dependent, increase of lactoferrin and IL-8 secretion within 24h in response to S. aureus or LPS. Marked increases (P < 0.05) in lactoferrin, TNF-alpha and serum amyloid A (SAA) mRNA expression were also observed. The results indicate the usefulness of our model to study infection responses of mammary epithelial cells, where all cells are simultaneously exposed to the same infection pressure. These responses can be studied over time, and most importantly, biological replication is provided by the four different genotypes being investigated individually. Finally, the results indicate that mammary epithelial cells play an important role in inflammatory response, through the production of pro-inflammatory cytokines, an acute phase protein, and lactoferrin. PMID- 15350750 TI - Total serum immunoglobulin M levels are affected by immunomodulators in seabream (Sparus aurata L.). AB - Immunoglobulin M (IgM) is a major component of the teleost humoral immune system. Despite the significance of IgM levels as an immune parameter, there are relatively few studies on changes induced in its total levels in serum. This study examines the effects of several immunomodulators (vitamin A, chitin, yeast cells or levamisole, which act as immunostimulants, and crowding, hypoxia or anaesthetics, which act as stressors) upon the total serum IgM levels of non immunized gilthead seabream (Sparus aurata L.). Total serum IgM levels of fish fed with the assayed immunostimulant-supplemented diets were statistically higher than those in fish fed a non-supplemented diet, especially in the case of levamisole. On the other hand, serum IgM levels of fish subjected to different stressors were not affected by crowding, hypoxia or certain anaesthetics. However, benzocaine and a narcotic dose of 2-phenoxyethanol provoked a great reduction, while quinaldine sulphate increased IgM levels to a significant degree. These results show how the seric IgM levels can be differently affected by some immunomodulators and the important role they may play in the regulation of total circulating IgM levels in seabream. The possibility of using total serum IgM for assessing immunostimulation, disease diagnosis and stress symptoms during fish farming is discussed. PMID- 15350751 TI - CpG oligodeoxynucleotides stimulate immune cell proliferation but not specific antibody production in rainbow trout (Oncorhynchus mykiss). AB - Bacterial DNA and CpG ODN have both been shown to have immunostimulatory effects in mammals, activating APCs and inducing a potent Th1 type immune response. They have also been shown to have a strong adjuvant effect and up-regulate MHC class 2 expression in murine cells, augment human and murine NK cell lytic activity, activate human B cells and induce murine B cell proliferation. However, little work has been carried out with regard to their effects on the piscine immune system. Here it is shown that various CpG ODN induce proliferation of peripheral blood leucocytes, spleen and head kidney cells from rainbow trout although, at the range of concentrations tested CpG ODN 2133 lacked the ability to induce specific antibody production to a protein antigen. PMID- 15350752 TI - Molecular cloning of the swine IL-4 receptor alpha and IL-13 receptor 1-chains: effects of experimental Toxoplasma gondii, Ascaris suum and Trichuris suis infections on tissue mRNA levels. AB - IL-4 and IL-13 are multi-functional cytokines with overlapping roles in the host defense against infection. Equally important in the regulation of IL-4 and IL-13 are their associated receptors. Though, their functional receptor complexes and signaling pathways are intricate and in some cases, share common elements, the specificity of the responses, nonetheless, resides in the structure and binding of the alpha-chain components. This report presents the cloning of the swine receptors IL-4Ralpha and IL-13Ralpha1 and the effects of parasite infection on their transcription. Pairwise alignment of predicted amino acid sequences indicates that the swine IL-13Ralpha1 is 86, 83, and 72% similar to canine, human and mouse sequences, respectively. Amino acid sequence conservation is appreciably lower between the swine IL-4Ralpha sequence and those from equine (72%), human (66%), and mouse (49%); however, noteworthy similarities were observed in their overall predicted secondary structures predominantly among the swine, equine, and human homologues. Relative levels of receptor mRNA in tissues from swine experimentally infected with the protozoan, Toxoplasma gondii (T. gondii) or the nematodes Ascaris suum (A. suum) or Trichuris suis (T. suis), which are known to induce Th1 or Th2 host responses, respectively, were measured by real-time PCR. Results indicated that within 14 days following infection, overall mRNA levels for IL-4Ralpha and IL-13Ralpha1 were elevated in T. gondii infected animals and reduced in A. suum-infected animals. Levels of swIL-4Ralpha and swIL-13Ralpha1 mRNA in T. suis-infected animals varied coincidentally with the course of the infection and the location of the analyzed tissue. PMID- 15350753 TI - Surfactant protein D expression in normal and pneumonic ovine lung. AB - Surfactant protein D (SP-D) is a collagenous calcium-dependent lectin constitutively expressed by alveolar type II pneumocytes and non-ciliated bronchiolar epithelial (Clara) cells. It binds to surface glycoconjugates expressed by a wide variety of microorganisms such as Gram-negative bacteria, influenza A virus, and various fungi, leading to pathogen inactivation or enhanced neutrophil and macrophage activity. Since a hallmark of bronchopneumonia is the initiation of inflammation in the bronchi and bronchoalveolar junction, we chose a classic ruminant model of bronchopneumonia caused by Mannheimia haemolytica to study the expression of SP-D within the bronchioles of infected lambs. Healthy weaned lambs were inoculated with either pyrogen-free saline (controls) or M. haemolytica intrabronchially using a fiber-optic bronchoscope. SP-D protein and mRNA expression in lung was detected by immunohistochemistry (IHC) and fluorogenic real-time relative quantitative reverse transcriptase polymerase chain reaction (real-time RT-PCR), respectively, during acute (1 day), subacute (15 days), and chronic (45 days) bronchopneumonia. At 15 and 45 days post-inoculation, areas of lung had peribronchiolar inflammatory cell infiltrate, epithelial cell hyperplasia, tortuosity of the airway lumens, and decreased intensity of SP-D protein staining and number of positive cells. The levels of SP D mRNA were not increased or significantly altered by M. haemolytica infection when compared to control animals. In conclusion, cell-associated SP-D protein expression significantly decreases within hyperplastic epithelium of lungs from infected animals during chronic bronchopneumonia. Exhaustion of SP-D protein reserves and absence of SP-D gene upregulation during the progression of bacterial pneumonia into chronicity may result in failure to clear the pathogen from the lung and/or cause animals to be more susceptible to re-infection. PMID- 15350754 TI - IL-4 induced CD23 (FcepsilonRII) up-regulation in equine peripheral blood mononuclear cells and pulmonary alveolar macrophages. AB - The objectives of this study were to quantify the induction of equine CD23 transcripts in equine peripheral blood mononuclear cells (PBMCs) and pulmonary alveolar macrophages cultured with recombinant equine IL-4 (rEq IL-4). PBMCs were isolated from blood drawn from four healthy horses. Bronchoalveolar lavage (BAL) fluid was collected from three healthy horses and alveolar macrophages were purified using adherence to plastic for 120 min. PBMCs and alveolar macrophages were cultured using four different conditions: rEq IL-4 and LPS, LPS alone, rEq IL-4 alone and a media control. Total RNA was isolated from cells cultured for 24 or 48 h. Reverse transcribed mRNA was amplified and quantified in real-time polymerase chain reaction (RT PCR) using a fluorescein labeled internal TaqMan probe for CD23 expression. Without exception, the relative value for CD23 mRNA transcripts from equine PBMCs and pulmonary alveolar macrophages cultured with rEq IL-4 for 24 and 48 h were higher than those cultured with LPS alone or the untreated control. Furthermore, morphologic changes were noted in alveolar macrophages cultured with rEq IL-4 prompting an investigation of cytokine expression levels. Alveolar macrophages cultured with LPS exhibited increased IL 8 and IL-12 p40 expression when compared to rEq IL-4, rEq IL-4 + LPS or the untreated control. These findings support two conclusions, (1) equine CD23 has a role in IL-4 mediated immune responses in the horse and (2) rEq IL-4 can modulate LPS-induced, pro-inflammatory cytokine production by equine pulmonary alveolar macrophages. PMID- 15350755 TI - Salmonella enteritidis clearance and immune responses in chickens following Salmonella vaccination and challenge. AB - Our previous work showed that the cell-mediated immunity (CMI) was enhanced by live Salmonella vaccine (LV). The objective of this study was to evaluate the impact of live and killed Salmonella vaccines on Salmonella enteritidis (SE) clearance and to determine if the clearance was mediated by cell-mediated and/or humoral immunity. Chickens were first immunized at 2 weeks of age followed by a booster dose at 4 weeks, challenged with live SE 2 weeks later (6-week-old) and tested for CMI, antibody response and SE clearance 1-week post SE-challenge (7 week-old). Spleen cell proliferation induced by SE-flagella and Concanavalin A (Con A) were significantly higher and SE shedding was significantly lower in the LV group. The splenic CD3 population was significantly lower and B cells were higher in the control group compared to all the SE-challenged groups (with and without vaccination). Serum antibody to SE-flagella and envelope were significantly higher in the KV group compared to all the other groups. These results suggest that LV protects against SE infection, probably by enhancing the CMI. PMID- 15350756 TI - Discovery and characterization of two types of liver-expressed antimicrobial peptide 2 (LEAP-2) genes in rainbow trout. AB - The sequences and gene organisation of two LEAP-2 molecules (LEAP-2A and LEAP-2B) from rainbow trout, Oncorhynchus mykiss are presented. Both genes consist of a 3 exon/2 intron structure, with exon sizes comparable to known mammalian genes. LEAP-2A notably differs from LEAP-2B in having larger introns and a larger 3' UTR. The predicted proteins contain a signal peptide and prodomain, followed by a mature peptide of 41 aa containing four conserved cysteines. The RXXR cleavage site to release the mature peptide was also conserved. Both genes were found to be constitutively expressed in the liver, with expression in the intestine, and to a lesser extent the skin, evident after bacterial challenge. PMID- 15350757 TI - Tryptic peptides of canine thyroglobulin reactive with sera of patients with canine hypothyroidism caused by autoimmune thyroiditis. AB - Canine thyroglobulin (cTg) was treated with trypsin at a ratio of trypsin to cTg of 1:100 (w/w). Tryptic peptides of cTg were analysed by Western immunoblotting for their reactivity to serum thyroglobulin autoantibodies (TgAA) from patients with TgAA-positive hypothyroidism and normal individuals. The sera of patients with TgAA-positive hypothyroidism reacted with several peptides: 43, 32.5 and 31 kDa; the sera of normal individuals did not bind these tryptic peptides. Some of the TgAA-positive sera of patients reacted with 25 kDa peptide in addition to three tryptic peptides above. This experiment was the first report about antigenic epitopes of cTg. These small tryptic peptides recognized by TgAA may be related with the induction of TgAA and may be useful as markers for autoimmune thyroid diseases in dog. PMID- 15350758 TI - Exploring the archives. PMID- 15350759 TI - At the margin: continuing crisis in British environmental history? AB - An understanding of historical relations between gender and nature, whether within the context of 'untouched wilderness' or the pulsating vibrancy of the industrial city, is central to collective self-knowledge in the world in which we all live. However, have British historians been less successful than their American counterparts in engaging with key aspects of the environmental past? PMID- 15350760 TI - Curicatures. AB - Marie Curie challenged many conventions when she left Poland, became a world famous French scientist and took X-ray machinery to the front line in World War I. Although she was the first person to win two Nobel prizes, many critics found it hard to believe that she could simultaneously be a caring mother and a brilliant researcher. As mythological accounts of Curie's success grew, photographs and caricatures of her reinforced traditional stereotypes of male and female roles in science. PMID- 15350761 TI - Sherlock Holmes: scientific detective. AB - Sherlock Holmes was intended by his creator, Arthur Conan Doyle, to be a 'scientific detective'. Conan Doyle criticized his predecessor Edgar Allan Poe for giving his creation - Inspector Dupin - only the 'illusion' of scientific method. Conan Doyle believed that he had succeeded where Poe had failed; thus, he has Watson remark that Holmes has 'brought detection as near an exact science as it will ever be brought into the world.' By examining Holmes' methods, it becomes clear that Conan Doyle modelled them on certain images of science that were popular in mid- to late-19th century Britain. Contrary to a common view, it is also evident that rather than being responsible for the invention of forensic science, the creation of Holmes was influenced by the early development of it. PMID- 15350762 TI - Fighting the 'microbe of sporting mania': Australian science and Antarctic exploration in the early 20th century. AB - The 'Heroic Age' of Antarctic exploration, which occurred during the first 15 years of the 20th century, captured headlines around the world. Australia was no exception, especially as Australian scientists played important roles in several expeditions. Through participation in the British Antarctic Expedition of 1907 1909, two Australian scientists - T.W. Edgeworth David (1858-1934) and Douglas Mawson (1882-1958) - became genuine national heroes, mainly through being members of the first party to reach the South Magnetic Pole area. At a superficial level, the vehicle of Antarctic exploration placed science at the forefront of public awareness, fulfilling David's ambition for greater recognition of science and scientists, especially considering the high level of public interest in sport. However, although David and Mawson gave Antarctic exploration a scientific veneer, simply through their status as scientists, the public viewed them as heroes because they had endured great hardships and conquered a point on the map in the name of science and the Empire. PMID- 15350763 TI - Shackleton's men: life on Elephant Island. AB - The experiences of the 22 men from Ernest Shackleton's Endurance expedition of 1914-1916 who were marooned on Elephant Island during the Antarctic winter are not as well known as the narrative of the ship being beset and sunk, and Shackleton's open boat journey to South Georgia to rescue them. Frank Wild was left in charge of the marooned men by Shackleton and saved them from starvation and despair. The morale of the men in the face of extreme exposure to the elements, the ingenuity of their devices for survival and their diet, conversation and entertainments all reveal heroic qualities of Shackletonian endurance. PMID- 15350764 TI - Learning from education to communicate science as a good story. AB - Science communicators must learn from science educators in their crusade to counteract the traditional boring and inefficient approaches to convey science. Educators encounter a need for methods of teaching that portray science as 'hard fun' and resources that encourage students' minds to burst into action. Narratives are considered by several authors as highly valuable resources for science education. However, little research has been undertaken to measure the efficiency of narratives in the context of science communication to the general public. Recent work however, suggests that narratives are indeed an alternative and an important means for science communication to convey information in an accurate, attractive, imaginative and memorable way. To present scientific information through stories, novels, comics and plays should be regarded as an important means to transmit information in the repertoire of both science teachers and science communicators. PMID- 15350765 TI - LINC: biology's revolutionary little computer. AB - The 1963 LINC (Laboratory INstrument Computer) stands at the center of two stories: the computerization of the biologist's laboratory and the advent of small-scale computing. The brainchild of Wesley Clark, 'the most brilliant computer designer of his generation', LINC was developed specifically to address the failure of biologists to adopt computer technology. To meet their unique needs, Clark built a machine the radical design of which defied and subverted the then dominant conventions of computer architecture. PMID- 15350766 TI - Understanding the current position of research in Africa as the foundation for child protection programs. PMID- 15350767 TI - Child abuse in South Africa: an examination of how child abuse and neglect are defined. AB - OBJECTIVE: The purpose of this study was to explore which of 17 categories of child maltreatment South Africans evaluated as most serious and to determine if those working with abuse and neglect evaluated abuse and neglect differently from those who did not. METHOD: A revised version of Giovannoni and Becerra's [Giovannoni, J., & Becerra, R. (1979). Defining child abuse. New York: The Free Press] questionnaire exploring the definition of abuse and neglect was completed by 181 residents of Cape Town, South Africa. The new form had 17 categories of child maltreatment, including 4 categories of societal abuse. Respondents were social workers (n = 57), human service workers (n = 42), laypersons (n = 65), and members of the child protection unit of the South African Police (n = 18). ANOVA was used to compare the groups' responses. When significant differences among groups were found, a Bonferroni post hoc test was run to determine differences between groups. RESULTS: The respondents ranked sexual abuse and child prostitution as most serious and housing and child labor as least serious of the 17 categories. There was a significant difference (p < or = .01) between groups on nine categories. When post hoc tests were run, differences were found for eight categories with laypersons generally evaluating categories as significantly more serious than social workers. CONCLUSIONS: Reasons for the order of the rankings are discussed, but concern remains that differences in the evaluation of child maltreatment will lead to difficulty in implementing a protocol for identifying and responding to incidents of abuse and neglect. PMID- 15350768 TI - Child sexual abuse in Tanzania and Kenya. AB - OBJECTIVE: Most research on child abuse in Tanzania and Kenya is unpublished in the international literature. The purpose of this paper is to examine the various commentaries and reports extant, toward an overview of the nature and frequency of child sexual abuse in Tanzania and Kenya. METHODS: Contacts were made with academics, government departments, NGOs, and UN agencies. This was followed by a field trip in the summer of 2001 where all available reports were examined and a wide range of interviews conducted. RESULTS: Little empirical data exist on child sexual abuse in Tanzania. It is widely perceived that it may be increasing as a result of AIDS sufferers' attempts to "cleanse" themselves. The breakdown of traditional childcare systems, foreign influences, poverty, and the lowly position of girls in society are also implicated. More research has been conducted in Kenya. It is clear that first coitus occurs at a young age for many Kenyan children and adolescents. Also, a degree of force, trickery, or material exchange is not uncommon in adolescent sexual relations. CONCLUSIONS: Child sexual abuse is under-researched in Tanzania and Kenya. Studies by UN agencies such as United Nations Children's Fund (UNICEF) and the International Labour Organisation (ILO) have focused on the commercial sexual exploitation of children, to the neglect of more pervasive abuse in children's own communities by family, relatives, and neighbors. Nationwide surveys of the general population are required for an empirical understanding of this topic. Given the high incidence of AIDS/HIV in both countries, it is important to know if the epidemic is increasing the risk of rape or incest for children. PMID- 15350769 TI - Attributions and discipline history as predictors of child abuse potential and future discipline practices. AB - OBJECTIVES: We attempted to identify factors that can be applied in primary and secondary prevention programs and expand the understanding of why those who were not abused may engage in abusive behavior. The purpose of this research was to explore how young adults' attributions of whether they deserved their childhood discipline, as well as their abuse history, relate to physical child abuse potential and their discipline plans for their future children. METHOD: A sample of 140 non-parent college students were asked to report on their discipline history, perceptions of that discipline, child abuse potential, and expected discipline practices. An age range of 18-20 was targeted for multiple reasons, including the suitability of these young adults for primary and secondary prevention programs. RESULTS: Analyses revealed that both physical child abuse potential and future discipline practices were independently predicted by respondents' belief that they deserved their discipline in conjunction with the harshness of their childhood discipline. DISCUSSION: These results suggest that the attributions of self-blame held by young adults about their discipline experiences are significant for increasing physical abuse potential regardless of whether the individual reports a history of abuse. PMID- 15350770 TI - Is there a specific relationship between childhood sexual and physical abuse and repeated suicidal behavior? AB - OBJECTIVE: Studies show that childhood sexual and physical abuse predict repeated suicide attempts and self-mutilation. Little is known about the importance of sexual and physical abuse when compared to other severe childhood adversities with respect to chronic suicidal behavior. METHOD: Seventy-four subjects, 65% of whom were women, consecutively admitted to a general hospital after having made a suicide attempt, were interviewed as part of the intake interview about prior suicide attempts and self-mutilation and received DSM-IV diagnoses. Sexual abuse, physical abuse, neglect, antipathy from parents, loss of parents, and severe discord in the family before the age of 18, were covered by the Childhood Experience of Care and Abuse (CECA) interview schedule. RESULTS: The prevalence of severe sexual abuse was 35%, severe physical abuse 18%, neglect 27%, antipathy 34%, loss of caregiver 37% and exposure to family violence 31%. Physical and sexual abuse were independently associated with repeated suicide attempts when controlling for the effects of the other childhood adverse factors. No other childhood adversity was related to chronic suicidal behavior. The odds ratio of exposure to sexual or physical abuse was highest among those who both repeated suicide attempts and self-mutilated. CONCLUSION: Physical and sexual abuse are significantly and independently associated with repeated suicidal behavior. PMID- 15350771 TI - Reactive attachment disorder in maltreated toddlers. AB - OBJECTIVE: To determine if Reactive Attachment Disorder (RAD) can be reliably identified in maltreated toddlers in foster care, if the two types of RAD are independent, and to estimate the prevalence of RAD in these maltreated toddlers. METHODS: Clinicians treating 94 maltreated toddlers in foster care were interviewed regarding signs of attachment disorder at intake in an intervention program. RESULTS: Using categorical and continuous measures, both types of RAD can be reliably identified in maltreated toddlers. Both continuous scores and categorical diagnoses indicated that a substantial minority of maltreated young children do exhibit signs of attachment disorders sufficient to meet criteria in DSM-IV and ICD-10. The two types were moderately convergent and at times co occurred in the same child. Prevalence of RAD in this high-risk sample was 38 40%. Indiscriminate/disinhibited RAD was identified in children with and without an attachment figure. Within this maltreated group, toddlers whose mothers had a history of psychiatric disturbance were more likely to be diagnosed with attachment disorders. CONCLUSIONS: RAD may be reliably identified in maltreated toddlers. Emotionally withdrawn/inhibited and indiscriminate/disinhibited types of RAD are not entirely independent. PMID- 15350772 TI - Factor structure and reliability of the Childhood Trauma Questionnaire in a Canadian undergraduate student sample. AB - OBJECTIVE: The aims of this study were to examine (1) the psychometric properties of the Childhood Trauma Questionnaire [CTQ; Bernstein, D., Fink, L., Handelsman, L., Foote, J., Lovejoy, M., Wenzel, K., Sapareto, E., & Ruggiero, J. (1994). Initial reliability and validity of a new retrospective measure of child abuse and neglect. American Journal of Psychiatry, 151, 1132-1136; Bernstein, D., & Fink, L. (1993). Manual for the Childhood Trauma Questionnaire. Unpublished manuscript. Bronx, NY: VA Medical Center] in an undergraduate sample and (2) the prevalence of abuse and neglect in this sample. METHOD: Principal components analyses (PCA), coefficient alpha, and correlations were used to analyse data for 470 undergraduate students from a mid-western Canadian university. Cut-offs from a short-form of the CTQ [Bernstein, D., & Fink, L. (1998). Manual for the Childhood Trauma Questionnaire. New York: The Psychological Corporation] were used to estimate prevalence. RESULTS: PCA yielded a five-factor solution comprised of emotional, physical, and sexual abuse, as well as emotional, and physical neglect. All factors, except physical neglect, demonstrated good internal consistency and test-retest reliability. This factor structure largely replicated results reported for an adolescent clinical sample [Bernstein, D., Ahluvala, T., Pogge, D., & Handelsman, L. (1997). Validity of the Childhood Trauma Questionnaire in an adolescent psychiatric population. Journal of the American Academy of Child and Adolescent Psychiatry, 36, 340-348], with the exception of the physical neglect factor which was comprised of considerably different items. Rates of childhood trauma were largely comparable to those reported for community and student samples in more densely populated regions of North America. CONCLUSIONS: The CTQ appears to be a valid measure of abuse and neglect in student samples, although experiences are somewhat differentially identified as different types of abuse and neglect depending on age and patient versus nonpatient status. PMID- 15350773 TI - The influence of dispersant concentration on the pore morphology of hydroxyapatite ceramics for bone tissue engineering. AB - There is a clinical need for synthetic scaffolds that will promote bone regeneration. Important factors include obtaining an optimal porosity and size of interconnecting windows whilst maintaining scaffold mechanical strength, enabling complete penetration of cells and nutrients throughout the scaffold, preventing the formation of necrotic tissue in the centre of the scaffold. To address this we investigated varying slip deflocculation in order to control the resulting porosity, pore size and interconnecting window size whilst maintaining mechanical strength. Hydroxyapatite (HA) porous ceramics were prepared using a modified slip casting process. Rheological measurements of the HA slips were used to identify deflocculation conditions which resulted in changes in the cell and window sizes of the resulting ceramics. Sintered ceramics were characterised by X-ray diffraction (XRD) and scanning electron microscopy (SEM). Pore and window size distribution was determined by SEM. XRD analysis confirmed that the crystal structure remained HA after the sintering process. SEM showed that HA porous ceramics presented a highly interconnected porous network with average pore sizes ranging from 391+/-39 to 495+/-25 microm. The average window size varied from 73+/-5 to 135+/-7 microm. Pore diameters obtained were controllable in the range 200-500 microm. Window sizes were in the range 30-250 microm. The use of dispersant concentration allows pore and window size to be modified whilst maintaining control over porosity demonstrated by a porosity of 85% for seven different dispersant concentrations. The advantage of this approach allows the correlation between the rheological conditions of the slip and the resultant sintered ceramic properties. In particular, optimising the ceramic strength by controlling the agglomeration during the casting process. PMID- 15350774 TI - The effect of backbone structure on polycation comb-type copolymer/DNA interactions and the molecular assembly of DNA. AB - A series of comb-type copolymers comprised of various polycation backbones and dextran (Dex) side chains were prepared to study the DNA/copolymer interaction. While the cationic copolymers with a lower degree of dextran grafts maintained an ability to condense DNA molecules into a globule form those with a higher degree of dextran grafting interacted with DNA without inducing DNA condensation. The structural differences in cationic backbones diversely influenced DNA hybridization as evaluated by circular dichroism (CD) spectrometry and UV-melting analyses. The copolymer having a polyallylamine (PAA) backbone induced B-->A-type transformation of DNA duplex, whereas the copolymers having either alpha-poly(l lysine) (alpha PLL) or epsilon-poly(l-lysine) (epsilon PLL) backbone induced B- >C-type transformation. The PAA copolymer is the first example of the artificial polymer that induces B-->A-type transformation under physiologically relevant condition. UV-melting analyses of DNA strands indicated that the alpha PLL copolymers showed the highest stabilization efficacy toward poly(dA).poly(dT) duplex and poly(dA).2poly(dT) triplex without affecting reversibility of inter DNA association. Melting temperatures (T(m)) of the triplex increased from 38 degrees Celsius to 99 degrees Celsius by the addition of the alpha PLL copolymer with an appropriate grafting degree. While the PAA copolymers had higher density of cationic groups along the backbone than alpha PLL copolymers, these copolymers moderately increased T(m) of the DNA triplex. The PAA copolymer caused considerable hysteresis in thermal melting/reassociation processes. Note that the PLL copolymers increased T(m) of the DNA triplex and not the duplex, suggesting their potential as a triplex selective stabilizer. Chemical structures of the cationic backbones of the copolymers were characteristically affected on the copolymer/DNA interaction even if their backbones were surrounded by abundant side chains (> wt%) of dextran. The study suggested that tailor-made design of "functional polycounterion" is a strategy to engineer molecular assembling of DNA. PMID- 15350775 TI - Effects of incorporation of HA/ZrO(2) into glass ionomer cement (GIC). AB - Glass ionomer cements (GICs) are a class of bioactive cements that bond directly to bone. In this paper, a new bioactive hydroxyapatite (HA)/zirconia (ZrO(2)) filled GIC composite was developed to improve the biocompatibility and bioactivity of the GICs with the surrounding bone and connective tissues. Nano sized HA/30 wt% ZrO(2) powders were heat treated at 700 degrees Celsius and 800 degrees Celsius for 3 h to elucidate the influence of the crystallinity of composite powders on the performance of HA/ZrO(2)-GICs. The effects of different volume percentages of HA/ZrO(2) powders (4, 12, 28 and 40 vol%) substituted within GICs were investigated based on their microhardness, compressive strength and diametral tensile strength. The HA/ZrO(2)-GICs composite was soaked in distilled water for 1 day and 1 week before subjecting the samples to mechanical testing. Results showed that the glass and HA/ZrO(2) particles were distributed uniformly in the GIC matrix. The substitution of highly crystalline HA/ZrO(2) improved the mechanical properties of the HA/ZrO(2)-GICs due to the slow resorption rate for highly crystalline powders in distilled water. The mechanical properties of HA/ZrO(2)-GICs increased with increasing soak time due to the continuous formation of aluminium salt bridges, which improved the final strength of the cements. The compositions 4 and 12 vol% HA/ZrO(2)-GICs exhibited superior mechanical properties than the original GICs. The mechanical properties of HA/ZrO(2)-GICs were found to be much better than those of HA-GICs because ZrO(2) has the attributes of high strength, high modulus, and is significantly harder than glass and HA particles. Furthermore, ZrO(2) does not dissolve with increasing soaking time. PMID- 15350776 TI - Synthesis of novel biodegradable polyanhydrides containing aromatic and glycol functionality for tailoring of hydrophilicity in controlled drug delivery devices. AB - Aromatic diacids were synthesized from chlorinated triethylene or pentaethylene glycols and p-hydroxy benzoic acid. The diacids were acetylated with acetic anhydride to produce homopolymers of polyanhydrides for controlled release applications such as the stabilization of proteins and drugs. Adding ethylene glycol segments into the acid monomer allows tailoring of the hydrophobicity of the polyanhydrides, which in turn dictates the solubility of molecules within the polymer matrix and degradation rate of the polymer. The glycol containing polyanhydrides were characterized by NMR, GPC, DSC and dissolution testing. The release characteristics and mechanism of the new polyanhydrides were evaluated using hydrophilic and hydrophobic dyes. PMID- 15350777 TI - Characterization of aqueous dispersions of Fe(3)O(4) nanoparticles and their biomedical applications. AB - A newly developed non-polymer coated Fe(3)O(4) nanoparticles showing well dispersion were synthesized using Fe(II) and Fe(III) salt chemical coprecipitation with tetramethylammonium hydroxide (N(CH(3))(4)OH) in an aqueous solution. Transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared spectrometer (FT-IR), X-ray photoelectron spectrometer (XPS) and superconducting quantum interference measurement device (SQUID) measurements were employed to investigate the iron oxide properties. The resulting iron oxide particles were manipulated to be as small as 9 nm diameter in size. Based on FT-IR and X-ray photoelectron spectrometer results, it is suggested that the surfaces of the magnetite (Fe(3)O(4)) particles are covered with hydroxide (-OH) groups incorporated with (CH(3))(4)N(+) through electrostatic interaction. The in vitro cytotoxicity test revealed that the magnetite particles exhibited excellent biocompatibility, suggesting that they may be further explored for biomedical applications. NMR measurements revealed significantly reduced water proton relaxation times T1 and T2. The MR images of the nanoparticles in water, serum, and whole blood were investigated using a 1.5 T clinical MR imager. Significant reduction of the background medium signal was achieved in the T2-weighted and the T2*-weighted sequence especially in the serum and whole blood. Combining the advantage of MRI signal contrast, the non-polymer coated surface chemistry for distinct bioconjugation and the homogenous nanometer size for better controlled biodistribution, these preliminary experiments demonstrated the potential of the as-synthesized magnetite material in functional molecular imaging for biomedical research and clinical diagnosis. PMID- 15350778 TI - In vitro corrosion study by EIS of a nickel-free stainless steel for orthopaedic applications. AB - The electrochemical impedance spectroscopy (EIS) technique was used for the study of the electrochemical behaviour of Ni-free austenitic stainless steel for orthopaedic applications. Experiments were carried out using four different test solutions: (i) phosphate-buffered saline (PBS), (ii) minimum essential medium (MEM), (iii) MEM + 10% fetal calf serum (FCS), (iv) MEM + 10% fetal calf serum + L929 fibroblast cell line (Cell). Bode-phase spectra showed the presence of two maxima and were fitted with an equivalent circuit characterized by two parallel combinations (Resistance, Constant Phase Element). The (R(1), CPE(1)) branch was assigned to the inner compact passive film and the (R(2), CPE(2)) branch to the external porous film. The resistance of the inner film R(1), here directly related to the material's uniform corrosion resistance, raised with the immersion time and increased in the following order: PBS 0.05). If hearts were treated with isoproterenol (ISO, 10 nmol/l), males exhibited significantly poorer recovery of post-ischemic contractile function than females (male: 13.0 +/ 1.9%; female: 28.1 +/- 1.2%; P < 0.05), and a significantly higher [Na(+)](i) accumulation during ischemia (male: 218 +/- 8%; female: 171 +/- 2%; P < 0.05). This ISO-induced male/female difference in [Na(+)](i) accumulation or contractile function was blocked by the nitric oxide synthase inhibitor, N(omega)-nitro-l arginine methyl ester (1 micromol/l). Furthermore, in ISO-treated hearts, the Na(+)/K(+)-ATPase inhibitor, ouabain (200 micromol/l) did not abolish the male/female difference in [Na(+)](i) accumulation during I/R or functional protection. Thus the data show that the sex difference in the [Na(+)](i) regulation is mediated through a NO-dependent mechanism, and the difference in susceptibility to I/R injury appears to result from a difference in Na(+) influx. PMID- 15350848 TI - Mechanism of oleic acid-induced gap junctional disassembly in rat cardiomyocytes. AB - This study investigated the mechanism of oleic acid (OA) on gap junctions and identified the protein kinase C (PKC) isoforms involved in OA-mediated gap junction disassembly in cardiomyocytes. Control cardiomyocytes showed continuous staining of the plasma membrane at cell-cell contact areas using antibodies reacting with connexin 43 (Cx43). The spontaneous contraction rate of cultured cardiomyocytes was reduced in a time-dependent manner by OA. In addition, Cx43 expression at cell-cell junction decreased, suggesting the disassembly of gap junction. Staining for PKC and PKCalpha, which were shown to colocalize with Cx43, also decreased with increased duration of OA treatment. The effects of OA on these distributional changes at cell junctions were reversed by 24 h incubation in fresh culture medium devoid of OA. Immunoprecipitation assays confirmed the biochemical binding between Cx43 and PKC/PKCalpha, and this protein interaction was not affected by OA. This may provide the basis for simultaneous detachment of Cx and PKC/PKCalpha from the cell-cell junction to the cytosol upon OA stimulation. Western blot analysis showed that OA-induced Cx43 Ser368 phosphorylation, and that this effect could be blocked by cotreatment with the general PKC inhibitor, calphostin C, the PKC inhibitor, eV1-2, or the Src kinase inhibitor, PP1, but not by the PKCalpha inhibitor, Go6976. eV1-2 also prevented the OA-induced disassembly of gap junctions. Taken together, these data suggest that OA-induced Cx43 Ser368 phosphorylation is mediated by activation of PKC and Src kinase and might be responsible for OA-induced gap junctional disassembly. PMID- 15350849 TI - Changes in cyclic nucleotide phosphodiesterase activity and calmodulin concentration in heart muscle of cardiomyopathic hamsters. AB - Cyclic nucleotides (cAMP and cGMP) phosphodiesterase (PDE) activities and expression are altered in the cardiac muscle of cardiomyopathic heart failure, and PDE inhibitors improve the abnormal muscle condition through changing the cyclic nucleotide concentration. These observations prompted us to investigate the role of calmodulin (CaM) in the regulation of cyclic nucleotide PDE activities, and moreover to study the modulation of the PDE isozymes in heart failure, using cardiac muscles of cardiomyopathic hamster. The CaM concentrations in the heart muscle of the normal control and cardiomyopathic hamsters (each of three to four hamsters) varied with cell fraction and with the age of the animal. The CaM concentrations in the soluble fraction obtained from cardiomyopathic hamster tissue were significantly increased at 25 and 32 weeks of age (2.02 +/- 0.62 microg/mg protein (mean +/- S.E.), and 3.21 +/- 0.95) compared with that obtained from the control (0.60 +/- 0.04) or cardiomyopathic (0.95 +/- 0.12) hamsters at 8 weeks of age. The solubilized PDE isolated from the hamster heart muscle (three or four hamsters in each age) by column chromatography on diethylaminoethyl (DEAE)-cellulose revealed three peaks of activity, which may correspond to the isozymes of PDE classified recently, namely PDE I, II, and III. These three peaks of activity, particularly peak III, seen in the soluble fraction of cardiomyopathic hamster heart declined in proportion to the age of the animal compared with that of the control hamster heart. In the cGMP-PDE assay system, the concentration of CaM inhibitor W-7 required for 50% inhibition (IC(50)) of PDE I, II, and III peak activities was 140, 29, and 46 microM, respectively, suggesting that PDE II is more sensitive to W-7. These results suggest that alteration in these isozyme activities accompanied with changes of CaM concentration may influence the cardiac muscle contractility in cardiomyopathic hamster via changes of cyclic nucleotide concentration. PMID- 15350850 TI - Cytokine-induced nitric oxide inhibits mitochondrial energy production and induces myocardial dysfunction in endotoxin-treated rat hearts. AB - The mechanism responsible for cardiac depression in septic shock remains unknown. The present study examined whether nitric oxide (NO) overproduced by inducible NO synthase (iNOS) can inhibit aerobic energy metabolism and impair the myocardial function in endotoxin-treated rat hearts. Lipopolysaccharide (LPS) significantly decreased systolic blood pressure (BP) to 44% of control during the 48 h treatment. Hearts from control and LPS-treated rats were perfused in a Langendorff apparatus. After LPS injection, left ventricular (LV) developed pressure (LVDP) was significantly depressed, plasma NO2-/NO3- (NO(x)) concentration was markedly increased, and myocardial adenosine 5'-triphosphate (ATP), creatine phosphate (CrP), and the ratio of ATP/adenosine 5'-diphosphate were progressively decreased with time. Immunological examination showed a significant expression of iNOS protein in the LPS-treated myocytes. Aminoguanidine, an inhibitor of iNOS, significantly attenuated these LPS-induced functional and metabolic changes. Myocardial cyclic guanosine 3',5'-monophosphate (cGMP) content was significantly increased after LPS injection. Methylene blue, an inhibitor of soluble guanylate cyclase, blunted this increase in cGMP and significantly restored the LPS-induced contractile dysfunction 6 h after LPS injection. In addition, there was a significant negative correlation between LVDP and myocardial cGMP levels as well as a significant negative correlation between LVDP and plasma NO(x) levels. In contrast, 48 h after LPS injection, methylene blue no longer affected cardiac performance, and there was a significant positive correlation between LVDP and myocardial ATP content. Furthermore, the normalized activities (as a ratio of the citrate synthase activity) of mitochondrial NADH CoQ reductase, succinate-CoQ reductase, and ATPase, were significantly inhibited, and the swelling or disruption of mitochondria cristae was seen in the 48 h LPS treatment. These LPS-induced functional and morphological disorders in the mitochondria were significantly improved by aminoguanidine. The findings suggest that sustained production of NO by iNOS leads to contractile dysfunction via cGMP in the early stage, but that it can directly impair the mitochondrial function, lower myocardial energy production, and contribute significantly to the myocardial dysfunction in the later stage of septic shock. PMID- 15350851 TI - Angiotensin II signals mechanical stretch-induced cardiac matrix metalloproteinase expression via JAK-STAT pathway. AB - BACKGROUND: Mechanical stress induces many pathophysiological effects in cardiomyocytes. The objective of this study was to test the hypothesis that angiotensin II is a potential mediator of stretch-induced activation of matrix metalloproteinases (MMP). METHODS: Neonatal rat cardiomyocytes grown on a flexible membrane were cyclically stretched achieving up to 20% elongation at 60 cycles/min (using a vacuum). We explored the signaling pathways involved in cyclical stretch-induced expression of MMP-14 and MMP-2. RESULTS: Cyclical mechanical stretch significantly increased mRNA expression and protein synthesis for MMP-14 and MMP-2 from the 6(th) to 24(th) h. The increase in MMP-14 and -2 proteins after stretch was completely blocked after the pretreatment with losartan (an angiotensin II AT1 receptor antagonist, 200 nM) and AG-490 (a Janus kinase 2 tyrosine kinase inhibitor, 100 nM) but not with PD 98059 (an inhibitor of p42/p44 mitogen-activated protein kinase, 50 microM) or wortmannin (a phosphatidylinositol 3-kinase, 30 nM). By zymography, MMP-2 activity was increased by cyclical stretch that was significantly attenuated by losartan and AG-490. The mechanical strain also increased the immunohistochemical labeling of MMP-14 and -2 that was attenuated by adding losartan. Cyclical stretch increased the expression of STAT-1 that was abolished by pretreating with losartan or AG 490 (50 microM and 100 microM). CONCLUSION: These findings indicate that cyclical stretch induces MMP-14 and -2 expression in neonatal rat cardiomyocytes and that the induction is mediated by the angiotensin II-JAK-STAT1 pathway. PMID- 15350852 TI - Phosphorylation-status of phospholamban and calsequestrin modifies their affinity towards commonly used antibodies. AB - Phospholamban (PLB) and calsequestrin (CSQ) play important roles in sarcoplasmic reticulum Ca(2+) transport and storage in cardiac muscle. Specific antibodies have been frequently used to quantitate CSQ and PLB protein levels. Here we demonstrate that two of the commonly available anti-PLB antibodies, anti-PLB-2D12 and anti-PLB-A1, show lower reactivity to phosphorylated than dephosphorylated PLB. A custom anti-PLB antibody, generated using a peptide corresponding to amino acids 2-14, is not affected by the phosphorylation state of PLB. In contrast, anti-CSQ reacts less with dephosphorylated CSQ than with phosphorylated CSQ. All three commercially available antibodies tested in this study have been widely used to quantify PLB and CSQ expression, and the results are integrated in many publications. Our studies reveal that the phosphorylation status of PLB and CSQ can affect antibody reactivity and may lead to over- or underestimation of the relative protein content and erroneous interpretation of data. PMID- 15350853 TI - Cognitive behavior therapy vs exposure in vivo in the treatment of panic disorder with agoraphobia (corrected from agrophobia). AB - Seventy-three psychiatric outpatients with DSM-IV diagnosis of panic disorder with agoraphobia were assessed with a battery of independent assessor, self observation, self-report and behavioral measures before and after therapy, and at a 1-yr follow-up. They were randomly assigned to Exposure in vivo (E; n = 25), Cognitive Behavior Therapy (CBT; n = 26), or a Wait-list control (WLC; n = 22) and received 12-16 individual therapy sessions, once weekly. The treatments yielded significant improvements, both on panic/agoraphobia measures and on measures of general anxiety, depression, social adjustment and quality of life, which were maintained at follow-up. However, there were no significant differences between E and CBT. The three criteria of clinically significant improvement were achieved by 67% of the E-patients and 79% of the CBT-patients at post-treatment, and 74% and 76%, respectively, at follow-up. The conclusion that can be drawn is that adding cognitive therapy to exposure did not yield significantly better results than for exposure alone. PMID- 15350854 TI - Social cognitive theory of posttraumatic recovery: the role of perceived self efficacy. AB - The present article integrates findings from diverse studies on the generalized role of perceived coping self-efficacy in recovery from different types of traumatic experiences. They include natural disasters, technological catastrophes, terrorist attacks, military combat, and sexual and criminal assaults. The various studies apply multiple controls for diverse sets of potential contributors to posttraumatic recovery. In these different multivariate analyses, perceived coping self-efficacy emerges as a focal mediator of posttraumatic recovery. Verification of its independent contribution to posttraumatic recovery across a wide range of traumas lends support to the centrality of the enabling and protective function of belief in one's capability to exercise some measure of control over traumatic adversity. PMID- 15350855 TI - Factorial structure and diagnostic efficiency of DSM-IV criteria for avoidant personality disorder in patients with binge eating disorder. AB - This study examined the factorial structure and diagnostic efficiency of the DSM IV criteria for avoidant personality disorder (AVPD). Two hundred and twenty eight consecutive outpatients (181 females and 47 males) with a primary diagnosis of binge eating disorder were reliably assessed with diagnostic interviews. Internal consistency of AVPD criteria was good, as suggested by coefficient alpha of 0.87, the pattern of inter-item correlations (range 0.41 to 0.64), and the lack of changes in alpha if any criteria are deleted. Exploratory factor analysis revealed a one-factor solution (56% of variance) supporting the unidimensionality of the AVPD criteria. Diagnostic efficiency indices (conditional probabilities, total predictive power, and kappa coefficients) were calculated for each AVPD criterion, for the entire study group and separately by gender. Overall, the best inclusion criterion was 'fears being ridiculed,' which was also the best predictor overall. These psychometric findings did not differ by gender. The findings support certain important aspects of the AVPD diagnosis. PMID- 15350856 TI - Measurement of compulsive hoarding: saving inventory-revised. AB - Four studies examined a new measure of compulsive hoarding (Saving Inventory Revised; SI-R). Factor analysis using 139 hoarding participants identified 3 factors: difficulty discarding, excessive clutter, and excessive acquisition. Additional studies were conducted with hoarding participants, OCD participants without hoarding, community controls and an elderly sample exhibiting a range of hoarding behavior. Internal consistencies and test-retest reliabilities were good. The SI-R distinguished hoarding participants from all other non-hoarding comparison groups. The SI-R showed strong correlations with other indices and methods of measuring hoarding (beliefs, activity dysfunction from clutter, observer ratings of clutter in the home) and relatively weaker correlations with non-hoarding measures (positive and negative affect and OCD symptoms). The SI-R appears to be an appropriate instrument for assessing symptoms of compulsive hoarding in clinical and non-clinical samples. PMID- 15350857 TI - Selective attention to threat in the dot probe paradigm: differentiating vigilance and difficulty to disengage. AB - The dot probe task [J Abnorm Psychol 95 (1986) 15] is an often-used paradigm to investigate selective attention to threat. A facilitated response to probes that appear at the same location of threat information in comparison with responses to probes at the opposite location of threat information is interpreted as vigilance for threat. We argue that the findings in the dot probe paradigm are ambiguous evidence for the vigilance to threat hypothesis. Results can also be interpreted as a difficulty to disengage from threat. In this study, 44 undergraduates performed a pictorial version of the probe detection task. Taking into account the reaction times on neutral trials, we found no evidence for a facilitated detection of threatening information. It was found that the dot probe effects are at least partially due to disengagement effects. The implications of these results for the understanding of attentional bias in normal and anxious individuals are discussed. PMID- 15350858 TI - The phobic stimuli response scales: a new self-report measure of fear. AB - Data from two large undergraduate samples were used to develop the phobic stimuli response scales (PSRS). Factor analyses of data from the first sample (N = 380) led to the creation of several scales, two of which (Social and Blood-Injection Fears) demonstrated good reliability and convergent validity. Analyses of data from a second sample (N = 330) were used to modify the three remaining scales (Animal, Bodily Harm, and Physical Confinement Fears), which also showed sound psychometric properties. As predicted, the PSRS were differentially correlated with neuroticism and extraversion, which may represent predisposing factors for these phobic responses. These new scales improve our understanding of the individual differences that are associated with various fears. PMID- 15350859 TI - Psychometric properties of the Thought-Action Fusion Scale in a Turkish sample. AB - The aim of the present study was to reveal the cross-cultural utility of the Thought-Action Fusion Scale (TAFS; J. Anxiety Disord. 10 (1996) 379). Thought action fusion (TAF) refers to the tendency to overvalue the significance and the consequences of thoughts. Two hundred and fifty one undergraduate Turkish students participated in the current study. The reliability and validity analyses of the Turkish version of the scale indicated that the TAFS had adequate psychometric properties in a Turkish sample. Consistent with the original TAF, the Turkish version of TAFS revealed two subscales as TAF-Likelihood and TAF Morality. Reliability analysis showed that TAF Scale and its factors had adequate internal consistencies and split-half reliability coefficients. Confirming the expectations, TAFS scores were found to be significantly and positively correlated with obsessive-compulsive symptoms, responsibility, and guilt measures. Moreover, it was found that people with high obsessive-compulsive symptoms had higher TAFS scores than those with low symptoms. PMID- 15350860 TI - Beliefs in personality disorders: a test with the personality disorder belief questionnaire. AB - The hypothesis that each personality disorder (PD) is characterized by a specific set of beliefs was tested in a sample of 643 subjects, including non-patient controls, axis-I and axis-II patients, diagnosed with SCID-I and -II interviews. Beliefs of six PDs (avoidant, dependent, obsessive-compulsive, paranoid, histrionic, borderline) were assessed with the Personality Disorder Belief Questionnaire (PDBQ). Factor analyses supported the existence of six hypothesized sets of beliefs. Structural equation modeling (SEM) supported the hypothesis that each PD is characterized by a specific set of beliefs. Path coefficients were however in the medium range, suggesting that PDs are not solely determined by beliefs. Nevertheless, empirically derived cutoff scores of the six belief subscales were reasonably successful in classifying subjects, percentages ranging form 51% to 83%. It appeared that there was a monotonical increase in scores on each belief subscale from non-patient controls, to patients without any PD, to patients with PDs (other than the pertinent PD), to patients with the pertinent PD. This suggests that PD-related beliefs are at least partly associated with (personality) psychopathology in general. Another explanation is that many patients' position on the underlying dimensions is not high enough to lead to a DSM PD diagnosis, but high enough to lead to an elevated belief score. PMID- 15350861 TI - The development of Plasmodium falciparum in experimentally infected Anopheles gambiae (Diptera: Culicidae) under ambient microhabitat temperature in western Kenya. AB - The effect of microhabitat temperature variation on the early development of Plasmodium falciparum in experimentally infected Anopheles gambiae s.s. (Diptera: Culicidae) was studied. Batches of mosquitoes were fed artificially on gametocyteamic blood obtained from human volunteers and then held in five environmental conditions described as: (1) incubator maintained at constant temperature of 28 +/- 1 degrees C as control; (2) temperature unregulated laboratory environment; (3) screen house; (4) grass thatched mud house and (5) corrugated iron roofed mud house. Both the grass and iron roofed mud houses were real houses found in the village communities around the ICIPE Research Centre in Mbita Point, Suba District south-western Kenya. The temperature and relative humidity of these holding environments were recorded over the study period. Mosquitoes were dissected after 24 h and 7 days to enumerate ookinetes and oocysts stages, respectively in their midguts. The mean temperature observed in the temperature-unregulated laboratory (28 degrees C) was significantly higher than the temperature of the screen house (24 degrees C) while the mean temperature observed in the iron roof mud house (27 degrees C) was comparable with that in the grass-thatched mud house (27 degrees C) although the iron roof house experienced more variation (coefficient of variation, C.V., = 9.6%) and higher peaking temperatures than the grass-thatch house. The mean relative humidity for the laboratory and screen house were 23% and 32.5%, respectively, much lower than relative humidity in the incubator (73%). Relative humidity of the grass thatch hut (42%) and Iron roof hut (51%) were also lower than those of the incubator. The ookinete intensities for mosquitoes in the screen house (10.11 +/- 1.79 ookinetes/midgut) were not statistically different (P = 0.41) from those held in the laboratory (7.50 +/- 1.19 ookinetes/midgut) or in the incubator (9.89 +/- 1.47 ookinetes/midgut). The holding environments influenced the oocyst infection rates (P = 0.04) that increased from 8.4% in the screen house to 10.2% in the laboratory. The highest infection rate (12.5%) was observed in mosquitoes held in the incubator. However, the mean oocyst intensities in mosquitoes did not differ under these environments (P = 0.58). In the 'real village house' environments, the mean ookinete intensities were not statistically different between groups of mosquitoes compared to the incubator (P = 0.86). The oocyst infection rates observed in the highly fluctuating iron roof house were 9.4% as compared to 9.0% and 6.9% in the more stable and constant habitats of grass thatch house and incubator, respectively. Results show that the natural microhabitats did not influence the infections rates in mosquitoes (P = 0.62). These findings indicate that the variation in temperatures prevailing in western Kenya particularly inside the village houses do not impede the development of malaria parasites in A. gambiae mosquitoes. PMID- 15350862 TI - Laboratory and field evaluation of Teknar HP-D, a biolarvicidal formulation of Bacillus thuringiensis ssp. israelensis, against mosquito vectors. AB - Larvicidal efficacy of Teknar HP-D, an improved biolarvicidal formulation of Bacillus thuringiensis ssp. israelensis (Bti), against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti was determined in the laboratory, and in field the efficacy of the formulation was tested against Cx. quinquefasciatus breeding in cesspits, unused wells and drains. The toxicity of the formulation to Gambusia affinis (larvivorous fish), Notonecta sp. and Diplonychus indicus (water bugs) was also evaluated in the laboratory. Teknar HP-D was field tested at three recommended dosages, 1, 1.5 and 2l/ha, selecting five habitats for each dosage. Another five habitats were kept untreated as controls. Ae. aegypti showed greatest susceptibility to the Bti toxin in the laboratory. In cesspits, all the three dosages caused >80% reduction of pupal recruitment up to day 6 post treatment, indicating that a weekly application at the lowest would be necessary for sustained control. The residual activity of the formulation was longer in unused wells, causing >80% reduction of pupal recruitment for 17 days from the day of treatment. In controlling pupal recruitment the three dosages produced equal effect. Application of Teknar HP-D at 1 l/ha once in three weeks is therefore recommended to control Cx. quinquefasciatus in unused wells. However, in drains, >80% reduction of pupal recruitment was observed for only 3 days and hence, application of Teknar HP-D at 2 l/ha that caused significantly higher level of reduction twice in a week at 3-day interval is necessary. At dosages from 0.032 to 3.2 mg/l (ppm), Teknar HP-D was non-toxic to Gambusia fish. The two predatory water bugs, Notonecta sp. and Diplonychus indicus that fed on the surviving larvae of Cx. quinquefasciatus exposed to the sub-lethal doses (LC(50) and LC(80)) of Teknar HP-D were safe with out having any mortality. PMID- 15350863 TI - Mortality profiles of Rhodnius prolixus (Heteroptera: Reduviidae), vector of Chagas disease. AB - Life table data of Rhodnius prolixus (Heteroptera: Reduviidae) kept at laboratory conditions were analysed in search for mortality patterns. Gompertz and Weibull mortality models seem adequate to explain the sigmoid shape of the survivorship curve. A significant fit was obtained with both models for females (R(2) = 0.70, P < 0.0005 for the Gompertz model; R(2) = 0.78, P < 0.0005 for the Weibull model) and for males (R(2) = 0.39, P < 0.0005 for the Gompertz model; R(2) = 0.48, P < 0.0005 for the Weibull model). The mortality parameter (b) is higher for females in Gompertz and Weibull models, using smoothed and non-smoothed data (P < 0.05), revealing a significant sex mortality differential. Given the particular life history of this insect, the non-linear relationship between the force of mortality and age may have an important impact in the vectorial capacity of R. prolixus as Chagas disease vector, and its consideration should be included as an important factor in the transmission of Trypanosoma cruzi by triatomines. PMID- 15350864 TI - Successful treatment of cutaneous leishmaniasis with lipid formulations of amphotericin B in two immunocompromised patients. AB - Pentavalent antimonial drugs are habitually the first choice for treating leishmaniasis, although they possess well-known toxicity and may present some therapeutic failure. Lipid formulations of amphotericin B (LFAB) have been increasingly used for treating several types of leishmaniasis. However, the administration of such lipid formulations specifically to patients with cutaneous leishmaniasis (CL) is still rare, including immunocompromised patients to whom standard treatments are more frequently contraindicated. We describe here two cases of immunocompromised patients with CL, one of them with AIDS, representing the first case of AIDS and CL co-infection treated with LFAB described in the literature. The patient achieved therapeutic success with a total 1.500 mg dose of amphotericin B colloidal dispersion. The other had diabetes mellitus as well as kidney failure and was under dialysis, having obtained the healing of lesion with a total dose of 600 mg of liposomal amphotericin B. Thus, the authors suggest that LFAB can represent a safe, efficient and less toxic therapeutic alternative to pentavalent antimonials, as well as to the so-called second line drugs, pentamidine and amphotericin B deoxycholate. PMID- 15350865 TI - Ultrasound organometry: the importance of body height adjusted normal ranges in assessing liver and spleen parameters among Chinese subjects with Schistosoma japonicum infection. AB - Hepatosplenic measurements among 550 Chinese subjects, aged 3-59 years from Yueyang city--a nonendemic area for schistosomiasis in Hunan province, China- were performed to define normal ranges of ultrasound organometry for assessing hepatosplenic morbidity in Schistosoma japonicum infection. Measurements included the size of the liver (left lobe and right lobe), the main portal vein stem, the peripheral periportal vein branches, and spleen length and thickness. The results document the significant relationship between body height and organometric parameters. The reference values stratified by body height improve the accuracy of assessment. Thus, height-based normal ranges established in this study can be applied in hospital routine and in field studies of patients infected with S. japonicum in Hunan province and in other endemic areas of China. PMID- 15350866 TI - Infection rate of Trypanosoma brucei s.l., T. vivax, T. congolense "forest type", and T. simiae in small wild vertebrates in south Cameroon. AB - In order to identify the infection rate of trypanosome species infecting wild animals in four localities (Bipindi, Campo, Fontem and Nditam) of southern Cameroon, 1,141 wild animals were sampled. These animals belonged to 36 species grouped in 8 orders including 407 primates, 347 artiodactyls, 264 rodents, 54 pangolins, 53 small carnivores, 11 saurians and crocodilians and 5 hyraxes. PCR using specific primers for Trypanosoma vivax, T. brucei s.l., T. congolense "forest type", and T. simiae showed that 18.7% of the animals were infected by at least one of these trypanosome species. A positive PCR result may not indicate absolutely an active infection because PCR can detect also transient infections. T. vivax (Duttonella) had the highest infection rate (9.5%) and was found in almost all the host orders studied. T. brucei s.l. mostly infected primates, rodents and some duikers (Cephalophus dorsalis and C. monticola). Trypanosomes of the subgenus Nannomonas had a lower infection rate of 5.5% (2.4% for T. simiae and 3.1% for T. congolense "forest type"). They were harboured mainly by primates, ungulates and rodents. Trypanosome infection rates were highest in Nditam (24.5%) and Bipindi (21%). T. brucei s.l. (Trypanozoon) had its maximum infection rate of 10.4% in Bipindi. The "Quantitative Buffy Coat" (QBC) and Kit for in vitro isolation techniques were used to identify 48 (6.1%) infected animals. 13 were positive using QBC, and 42 were positive by KIVI. However, PCR was negative on 16 of these infected animals, probably due to infections with other trypanosome species. This study showed that trypanosomes of the subgenera Duttonella, Nannomonas and Trypanozoon could infect small wild vertebrates as has been shown for large ungulates and carnivores. The presence of T. brucei s.l. in a large range of wild animals strengthens the hypothesis of the existence of a wild animal reservoir of T. b. gambiense in Cameroon. PMID- 15350867 TI - Serological surveillance of brucellosis and Q fever in cattle in the Central African Republic. AB - No data are available concerning the seroprevalence of brucellosis in Central African Republic (CAR) and the last report concerning the seroprevalence of Q fever in CAR is from 1995. The aim of our study was to determine the prevalence of these diseases in CAR, especially in Zebu cattle. We used the Rose Bengal Plate Test to test 2032 bovine serum samples for antibodies to Brucella spp. and an indirect immunofluorescence assay to test 784 bovine serum samples for antibodies to Coxiella burnetii (the species responsible for Q fever). The mean seroprevalences of antibodies to Brucella and Coxiella were 3.3 and 14.3%, respectively. Significant differences were found between regions and herds for both diseases. However, relation with differences of climate or vegetation were not evident. Therefore, further data are necessary to better understand the epidemiology of these diseases in CAR and evaluate losses to the farmers. PMID- 15350868 TI - Cystic echinococcosis in Argentina: evolution of metacestode and clinical expression in various Echinococcus granulosus strains. AB - Echinococcus granulosus hydatid cysts were examined in 41 patients from Neuquen and Tucuman provinces in Argentina. Sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) revealed in 19 patients common sheep strain (G1), in 6 patients Tasmania sheep strain (G2), in 1 patient cattle strain (G5), and in 15 patients camel strain (G6). In Argentina the only known is the domestic cycle that affects dogs and herbivorous, including ovine, swine, cattle and goats. These strains produced a total of 58.6% of primary liver infections, 29.2% primary in lung, 2.4% primary in spleen and 9.8% were multiorgan abdominal infections. The metacestode was classified using the evolutive stages proposed by WHO-IWGE (from CE1 to CE5). We estimated that CE1 cyst has a duration of about 22 years, CE2 of 14 years, CE3 of 10 years, CE4 of 19 years and CE5 was not determined. The active types CE1 and CE2 reached 75% of all cases from all strains. In 36 patients with cysts from G1, G5 and G6 strain, there were only two asymptomatic cases. The strains of the E. granulosus complex do not present important clinical differences; only G6 seems to have higher growth rate. PMID- 15350869 TI - Human fasciolosis in Van province, Turkey. AB - This investigation was carried out to determine the frequency of the fasciolosis in Ercis, a town in the Van province located in the Eastern Turkey. The study includes 500 asymptomatic subjects (322 males and 178 females) whose ages ranged from 5 to 75 years. In all subjects, stool samples were examined at initiation of the study and one week later to rule out pseudoparasitosis. The methods of flotation (in saturated saline solution) and sedimentation (in formalin-ether solution) were used in examination of the stool samples. Fasciola hepatica eggs were detected in nine (1.8%) out of 500 subjects. In conclusion, our data showed that the prevalence of human fasciolosis was high in our region because watercress consumption was common. We also think that human fasciolosis is more frequent than expected in Turkey. To determine the exact prevalence of the disease, extensive serological investigations should be performed in different regions of Turkey. PMID- 15350870 TI - Freeway safety as a function of traffic flow. AB - In this paper, we present evidence of strong relationships between traffic flow conditions and the likelihood of traffic accidents (crashes), by type of crash. Traffic flow variables are measured using standard monitoring devices such as single inductive loop detectors. The key traffic flow elements that affect safety are found to be mean volume and median speed, and temporal variations in volume and speed, where variations need to be distinguished by freeway lane. We demonstrate how these relationships can form the basis for a tool that monitors the real-time safety level of traffic flow on an urban freeway. Such a safety performance monitoring tool can also be used in cost-benefit evaluations of projects aimed at mitigating congestion, by comparing the levels of safety of traffic flows patterns before and after project implementation. PMID- 15350871 TI - Sensitivity to punishment and sensitivity to reward and traffic violations. AB - The aim of our study consists of contributing information on the relationship between the personality variables derived from Gray's model and the conduct that accompanies the infringement of the road traffic rules. Seven hundred and ninety two adults of both sexes took part in the study (389 men and 403 women), all of whom had driving licences and drove frequently. The subjects answered "The Sensitivity to Punishment and Sensitivity to Reward Questionnaire", a scale of monotony avoidance, and two Likert scales of attitude and behaviour in connection with traffic violations. We found a high positive relationship between attitude and behaviour, with the men infringing the rules more than the women. Hypotheses regarding a relationship between traffic offences and sensibility to reward and monotony avoidance were confirmed. Those people with high scores in sensitivity to punishment and low ones in sensitivity to reward were those who drove within the law, while those with low sensitivity to punishment and high sensitivity to reward were those who broke it more. Sensitivity to reward was a stronger determinant in encouraging infringement of the rules than was sensitivity to punishment in discouraging the subjects to do so. PMID- 15350872 TI - Ranking and selection of motor carrier safety performance by commodity. AB - We use recent safety performance data to rank US motor carrier commodity segments (e.g., Tank segment or Produce segment) in terms of several driver-related, vehicle-related, and crash-related safety measures. Ranking and selection inference techniques are used to determine the best and worst performing commodity segments at the 95% confidence level. The results are mixed, however the Passenger segment is generally best, while the Produce, Intermodal, and Refrigerated segments tend to be worst. PMID- 15350873 TI - Comparison of reporting of seat belt use by police and crash investigators: variation in agreement by injury severity. AB - PURPOSE: To evaluate agreement between police and trained investigators regarding seat belt use by crash victims, according to injury severity. METHODS: We used data from the National Accident Sampling System Crashworthiness Data System (CDS) for front seat occupants, 16 years and older, in crashes during 1993-2000. Crashworthiness Data System investigators determined belt use from vehicle inspection, interviews, and medical record information; their assessment was considered the gold standard for this analysis. Occupant severity of injury was categorized in five levels from no injuries to death. We estimated the sensitivity, specificity, and area under receiver operating characteristic curves for police reports of belt use. RESULTS: Among 48,858 occupants, sensitivity of a police report that a belt was used was 95.8% overall and varied only modestly by injury severity. Specificity of a police report that a belt was not used was 69.1% overall; it was the lowest among the uninjured (53.2%) and greatest among the dead (90.4%). The area under the curve was 0.82 (95% confidence interval 0.82 0.83) overall; this was lowest among those not injured (0.75, 95% confidence interval 0.74-0.76) and increased with injury severity to 0.91 (95% confidence interval 0.90-0.93) among those who died. CONCLUSION: Police usually classify belted crash victims as belted, regardless of injury severity. But they often classify unbelted survivors as belted when they were not. This misclassification may result in exaggerated estimates of seat belt effectiveness in some studies. PMID- 15350874 TI - Rural and urban traffic fatalities, vehicle miles, and population density. AB - The purpose of this study was to evaluate the effect of population density on the rates of motor vehicle mortality in rural and urban areas, while controlling for vehicle miles traveled (VMT). Rural and urban data for traffic mortality, VMT, and population were obtained for each state from the Federal Highway Administration for 1998-2000. Linear regression was used to estimate the effect of population density, VMT per capita, southern location, and presence of a trauma system on mortality. Variation in rural mortality rate (per 100,000 population) was proportional to rural VMT per capita, but population density and southern location were also independent predictors, together accounting for 91% of this variation. Variation in urban mortality rates was not affected by population density, but urban rates were also higher in the south. The exposure based rural mortality rate (deaths per 100 million VMT) was inversely proportional to population density, which along with southern location explained 41% of the variation from state to state. The presence of a state trauma system did not measurably affect mortality. After controlling for VMT and southern location, state population density was a moderately strong predictor of rural but not urban traffic mortality rates. PMID- 15350875 TI - A spatially disaggregate analysis of road casualties in England. AB - Spatially disaggregate ward level data for England is used in an analysis of various area-wide factors on road casualties. Data on 8414 wards was input into a geographic information system that contained data on land use types, road characteristics and road casualties. Demographic data on area-wide deprivation (the index of multiple deprivation) for each ward was also included. Negative binomial count data models were used to analyze the associations between these factors with traffic fatalities, serious injuries and slight injuries. Results suggest that urbanized areas are associated with fewer casualties (especially fatalities) while areas of higher employment density are associated with more casualties. More deprived areas tend to have higher levels of casualties, though not of motorized casualties (except slight injuries). The effect of road characteristics are less significant but there are some positive associations with the density of "A" and "B" level roads. PMID- 15350876 TI - Estimating the relationship between accident frequency and homogeneous and inhomogeneous traffic flows. AB - This paper estimates the relationship between accident frequency and the traffic flow empirically treating the hourly traffic flow in two different ways, as consisting of homogenous vehicles and as consisting of cars and lorries. Rural roads in Sweden are studied using Poisson and Negative Binominal regression models. It is found that important information is lost if no consideration is taken to differences between vehicle types when estimating the marginal effect of the traffic flow. The accident rate decreases when the traffic flow is treated as if homogeneous. However, when cars are studied separately the result suggests that the accident rate is constant or increases. The result with respect to lorries is reversed, indicating a decreasing number of accidents as the number of lorries increases. PMID- 15350877 TI - Some consequences of different older driver licensing procedures in Australia. AB - Given both the expected growth in the number of older drivers and their over involvement in fatal and serious injury crashes, there has been a world-wide call for improved licensing procedures to manage older driver safety. In particular, licensing authorities have been urged to move from mandatory assessment of all older drivers to assessment practices targeting only those at higher crash risk. The current study examined older driver fatal and serious injury crash involvement rates across all Australian States to determine a possible association with the different licensing procedures. In particular, older driver crash involvement rates in Victoria (where there is no age-based assessment program) have been compared with rates in other jurisdictions with assessment programs. Crash involvement rates have been calculated using two denominators: per population and per number of licensed drivers. Some data limitations notwithstanding, older drivers in jurisdictions with age-based mandatory assessment programs could not be shown to be safer than drivers in Victoria. Further, there is some indicative evidence that older drivers in Victoria may have a significantly safer record regarding overall involvement in serious casualty crashes. PMID- 15350878 TI - A two-step medically based injury surveillance system--experiences from the Oslo injury register. AB - This paper presents a two-step injury surveillance system. In the first step, limited data (a minimum data set) on all (or a representative sample of all) injuries to residents and non-residents within a defined geographical area were obtained using routine collection procedures within the medical care system. The second step involved periodically sampling of specific injuries, injured persons, or places for in-depth investigations from the database established by the first step, or selecting relevant injured persons seeking treatment in the medical care system, to collect many data (an expanded data set) on a limited number of injuries. This system was implemented in Oslo. Data from about 48,000 injuries were collected annually. Two in-depth investigations of serious occupational injuries were carried out. The first involved 223 cases and the second, 50 cases. Some in-site studies were included. Experiences from the implementation in Oslo suggest that this system can function in the medical care system and provide data required for making estimates of injury incidence rates, establishing trends, and on contributing factors to injuries. A crucial factor in the success of the first part of such a system is to have enough resources for continuous quality control and feedback to personnel involved in the registration of data. Combining the registered data from general practitioners, accident and emergency departments, hospitals and notifications of fatalities in Oslo, and on assessments of the number of injuries treated by private clinics and occupational health centres in Oslo, and in the health care system outside of Oslo, leads to the conclusion that 11.9% of the residents of Oslo will annually be treated for an injury. PMID- 15350879 TI - Bayesian spatial and ecological models for small-area accident and injury analysis. AB - In this article, recently developed Bayesian spatial and ecological regression models are applied to analyse small-area variation in accident and injury. This study serves to demonstrate how Bayesian modelling techniques can be implemented to assess potential risk factors measured at group (e.g. area) level. Presented here is a unified modelling framework that enables thorough investigations into associations between injury rates and regional characteristics, residual variation and spatial autocorrelation. Using hospital separation data for 83 local health areas in British Columbia (BC), Canada, in 1990-1999, we explore and examine ecological/contextual determinants of motor vehicle accident injury (MVAI) among male children and youth aged 0-24 and for those of six age groups (<1, 1-4, 5-9, 10-14, 15-19 and 20-24). Eighteen local health area characteristics are studied. They include a broad spectrum of socio-economic indicators, residential environment indicators (roads and parks), medical services availability and utilisation, population health, proportion of recent immigrants, crime rates, rates of speeding charge and rates of seatbelt violation. Our study indicates a large regional variation in MVAI in males aged 0 24 in British Columbia, Canada, in 1990-1999, and that adjusting for appropriate risk factors eliminates nearly all the variation observed. Socio-economic influence on MVAI was profoundly apparent in young males of all ages with the injury being more common in communities of lower socio-economic status. High adult male crime rates were significantly associated with high injury rates of boys aged 1-14. Seatbelt violations and excess speeding charges were found to be positively associated with the injury rates of young men aged 20-24. This and similar ecological studies shed light on reasons for regional variations in accident occurrence as well as in the resulting injuries and hospital utilisation. Thereby they are potentially useful in identifying priority areas for injury/accident prevention and in informing regional health planning and policy development. PMID- 15350880 TI - Effects of remote and in-person verbal interactions on verbalization rates and attention to dynamic spatial scenes. AB - This study focused on how teams allocated attention between a driving-related spatial task and a verbal task, and how different kinds of verbal interactions affected performance of the driving-related task. In Experiment 1, 29 two-person teams performed an interactive verbal task while one team member also performed a simulated driving task. Of the team members performing only the verbal task, half could see their partner's spatial situation, as a car passenger can (in-person condition), and half were remotely located, similar to someone speaking to a driver using a cell-phone. Teams interacted verbally at an overall slower rate during remote than in-person interactions, suggesting that remote verbal interactions are more difficult than in-person interactions. Verbal interactions degraded situation awareness for driving-related information while performing the spatial task; and this degradation was not greater during remote than in-person interactions. Experiment 2 used a faster-paced verbal task and found greater degradation of situation awareness due to the verbal task. These findings are potentially relevant to the issue of how passenger and cell-phone conversations affect driving performance. PMID- 15350881 TI - Wertheim's hypothesis on 'highway hypnosis': empirical evidence from a study on motorway and conventional road driving. AB - This paper aims to study the phenomenon known as 'highway hypnosis' or 'driving without attention mode', which has been defined as a state showing sleepiness signs and attention slip resulting from driving a motor vehicle for a long period in a highly predictable environment with low event occurrence, this being the case with motorways and very familiar roads [Highway hypnosis: a theoretical analysis. In: Gale, A.G., Brown, I.D., Haslegrave, C.M., Moorhead, I., Taylor, S. (Eds.), Vision in Vehicles-III. Elsevier, North-Holland, pp. 467-472]. According to Wertheim's hypothesis on 'highway hypnosis', long-term driving on motorways and conventional roads, e.g. main roads, secondary roads--implies differences in the predictability of the movement pattern of the visual stimulation, in the eye musculature activity and in the type of feedback used in visual information processing (mostly extra-retinal on motorways and retinal and extra-retinal on conventional roads). All this ultimately leads to alertness differences between both road types. Our research is intended to provide empirical evidence from the hypothesis, based on the data recorded during the actual driving experience of a group of subjects on a motorway and a conventional road. We studied whether or not significant alertness differences were found-measured by EEG data relative to time periods of on-target eye-tracking performance--between motorway and conventional road driving. Our results partially support the hypothesis, as drowsiness proved to be higher on motorways than on conventional roads during the final driving period but not during the starting stage, when the opposite trend was noticed. This result could be explained by the fact that during the first driving periods the effects of the stimulus movement predictability had not yet become apparent, since they tend to show after a long drive. PMID- 15350882 TI - Impact of infrastructure and local environment on road unsafety. Logistic modeling with spatial autocorrelation. AB - This article aims at modeling the impact of road characteristics and local spatial environment on road (un)safety. The study applies to Belgium where some 1,500 people are killed annually on the roads. This statistic corresponds to one of the highest risks in Europe. Road unsafety is expressed here as whether an hectometer of road belongs to a black zone; a black zone is defined as a segment of road where roads accidents are concentrated. Logistic modeling including spatial autocorrelation is used and compared to non-spatial regression. It is shown that a spatial model is needed to avoid biased estimated parameters. Results show that local environment and road infrastructure play a substantial role in the co-occurrence of road accidents. Hence, education and enforcement cannot be the only measures taken to reach a sustainable road safety. To attain their objectives of accident reduction, public authorities should also take their responsibilities in the matter of securing road infrastructure. PMID- 15350883 TI - Identifying predictors of persistent non-alcohol or drug-related risky driving behaviours among a cohort of young adults. AB - This study sought to identify adolescent risk factors that predicted persistent risky driving behaviours among young adults. It was part of a longitudinal study of a birth cohort (474 males and 459 females). The potential predictors were self reported data obtained at ages 15, 18, 21 years (academic qualifications, personality, mental health, anti-social behaviour and driving behaviour). The risky driving behaviour outcomes were obtained at ages 21 and 26 years and included driving fast for thrills, taking deliberate risks for fun, excessive speed, dangerous overtaking, and close following (tailgating). Persistent risky drivers were defined as those who often, or fairly often engaged in a behaviour at both ages. A minority of males and very few females were classified as persistent risky drivers. Among the males, the factors that predicted at least one, or more of the outcomes were the personality trait of low constraint (i.e. low scores for control, harm avoidance, and traditionalism), aggressive behaviour, and cannabis dependence. These are characteristics to be borne in mind when developing programmes for young drivers that aim to deter the development of persistent risky driving behaviour. PMID- 15350884 TI - Crash reduction following installation of centerline rumble strips on rural two lane roads. AB - Rural two-lane roads generally lack physical measures such as wide medians or barriers to separate opposing traffic flows. As a result, a major crash problem on these roads involves vehicles crossing the centerline and either sideswiping or striking the front ends of opposing vehicles. These types of opposing direction crashes account for about 20% all fatal crashes on rural two-lane roads and result in about 4,500 fatalities annually in the US. The present study evaluated a potential engineering countermeasure for such crashes-installation of rumble strips along the centerlines of undivided rural two-lane roads to alert distracted, fatigued, or speeding motorists whose vehicles are about to cross the centerlines and encroach into opposing traffic lanes. Data were analyzed for approximately 210 miles of treated roads in seven states before and after installation of centerline rumble strips. An empirical Bayes before-after procedure was employed to properly account for regression to the mean while normalizing for differences in traffic volume and other factors between the before and after periods. Overall results indicated significant reductions for all injury crashes combined (14%, 95% confidence interval (95% CI) = 5-23%) as well as for frontal and opposing-direction sideswipe injury crashes (25%, 95% CI = 6-44%)--the primary target of centerline rumble strips. In light of their effectiveness and relatively low installation costs, consideration should be given to installing centerline rumble strips more widely on rural two-lane roads to reduce the risk of frontal and opposing-direction sideswipe crashes. PMID- 15350885 TI - An explorative study of the relationship between lifestyle and driving behaviour among young drivers. AB - The high accident risk among young drivers is a well-known and well-documented fact in most countries, including Denmark. Lifestyle has proven to be related to driving behaviour as well as accident risk among young drivers. However, the underlying process through which the relationship between the lifestyle and the driving behaviour is established is not yet fully understood. Using focus group interviews divided by sex and education this study explores the psychosocial function of driving as well as the process through which a relationship between lifestyle and driving behaviour is established. Twenty-nine young drivers living in the Copenhagen area participated in the study. Data were analysed using a modified version of the Editing Analysis Style. PMID- 15350886 TI - Effects of lowering the legal BAC to 0.08 on single-vehicle-nighttime fatal traffic crashes in 19 jurisdictions. AB - BACKGROUND: In the past few decades, numerous policies, including those that lower legal blood alcohol concentration limits, have been enacted to reduce alcohol-impaired driving. In the US, 41 states and the District of Columbia have enacted 0.08 per se laws, which specify that if a driver's BAC is at or above 0.08, a violation has occurred even if the driver does not show signs of intoxication. OBJECTIVE: We examined effects of lowering the blood alcohol concentration limit to 0.08 per se on fatal traffic crashes in 18 states and the District of Columbia, and whether effects of the law varied by state or by baseline rates of fatal traffic crashes. METHOD: Data on fatal traffic crashes were obtained from the Fatality Analysis Reporting System, including all states that enacted 0.08 per se prior to 2001 in the contiguous United States. Effects of the 0.08 law were examined in each state separately, and the overall effect across states was examined using a mixed-model Poisson regression on single vehicle-nighttime fatal traffic crashes. RESULTS: State-specific analyses showed that fatal traffic crashes significantly decreased in three of the 19 states following the introduction of the 0.08 law, prior to adjusting for potential confounders. The mixed-model regression showed a statistically significant 5.2% reduction in single-vehicle-nighttime fatal traffic crashes associated with the 0.08 law across all states, after adjusting for administrative license revocation, the number of Friday and Saturday nights in a month, and trends in all other types of fatal traffic crashes. Findings indicate that the effect of the 0.08 law does not vary significantly by state or baseline rate of fatal traffic crashes in a state, and no significant statistical interaction exists between 0.08 and administrative license revocation policy effects. PMID- 15350887 TI - Long term medical costs of motor vehicle casualties in Alberta (1999): a population-based, incidence approach. AB - The purpose of this paper is to estimate the long term medical costs attributable to motor vehicle accidents (MVAs) for all persons in Alberta, Canada in 1999, primarily using observational data. Injury claims with personal identifiers for 1999 were reported by the automobile insurance companies. These records were linked to the provincial health registry which covers the entire population. The registry is linked to databases which identify all inpatient and outpatient (including emergency room) visits, physician services, and other health records. Utilization and costs were derived for all casualties who were admitted to hospital or seen in an emergency room, and for a large sample of other (low severity) cases; a sample of matched controls was derived and their costs were also estimated. Actual costs were obtained for 3 years, and longer term costs were projected for subsequent years. Total costs attributable to MVAs were estimated at over $117 million for 1999. Average net costs per casualty, reported by severity group, were: $22.9 thousand for hospital cases; $3.6 thousand for emergency room-only cases; and $157 for other cases. Long term costs were 65% of first year costs for hospitalized cases and 250% for emergency room cases. Overall, aggregate costs for all non-hospital cases exceeded those for hospitalized cases. PMID- 15350888 TI - Differences in safety belt use by race. AB - The purpose of the study was to identify daytime differences in safety belt use by race. Safety belt use was investigated in a direct observation survey of drivers and front-outboard passengers throughout Michigan. Data were weighted to calculate statewide safety belt use rates by race. Race was assessed visually by trained observers. The study showed that motor vehicle occupants identified as Black had significantly lower safety belt use than those occupants identified as White or Other. PMID- 15350889 TI - The burden of injuries in the Philippines: implications for national research policy. AB - Injuries cause 10% of the mortality and 15% of disability worldwide. However, there is a paucity of data on injuries in the developing world where two-thirds of all injury deaths occur. This is the first published report characterizing the overall problem of injuries in the Philippines, a developing country in southeast Asia. This report defines the burden of injuries in the Philippines and identifies priority areas for the national health research agenda. A systematic review of 35 years of published and unpublished data on injuries in the Philippines (1960-1995) was conducted. Injury fatality rates increased by 196% from 14.3 per 100,000 in 1960 to 42.3 per 100,000 in 1995, and one in 11 deaths in the Philippines are due to injuries. Intentional injuries account for 48% of all injury deaths and motor vehicle crashes for 15%. For 15-44 year old males, injuries account for 42% of all deaths, 67% of which are intentional. The proportion of all deaths attributable to intentional injuries has increased by 925% and that of motor vehicle crashes by 600% from 1960 to 1995. Improvements in injury surveillance and documentation of non-fatal injury outcomes are needed. Research into risk factors and potential interventions for the prevention of intentional injuries should be a priority in the Philippines. PMID- 15350890 TI - Possible aggregation biases in road safety research and a mechanism approach to accident modeling. AB - In accident reconstruction, individual road accidents are treated as essentially deterministic events, although incomplete information can leave one uncertain about how exactly an accident happened. In statistical studies, on the other hand, accidents are treated as individually random, although the parameters governing their probability distributions may be modeled deterministically. Here, a simple deterministic model of a vehicle/pedestrian encounter is used to illustrate how naively applying statistical methods to aggregated data could lead to an ecological fallacy and to Simpson's paradox. It is suggested that these problems occur because the statistical regularities observed in accident data have no independent status, but are simply the result of aggregating particular types and frequencies of mechanisms. PMID- 15350891 TI - Spatial analysis of alcohol-related motor vehicle crash injuries in southeastern Michigan. AB - Temporal, behavioral and social risk factors that affect injuries resulting from alcohol-related motor vehicle crashes have been characterized in previous research. Much less is known about spatial patterns and environmental associations of alcohol-related motor vehicle crashes. The aim of this study was to evaluate geographic patterns of alcohol-related motor vehicle crashes and to determine if locations of alcohol outlets are associated with those crashes. In addition, we sought to demonstrate the value of integrating spatial and traditional statistical techniques in the analysis of this preventable public health risk. The study design was a cross-sectional analysis of individual-level blood alcohol content, traffic report information, census block group data, and alcohol distribution outlets. Besag and Newell's spatial analysis and traditional logistic regression both indicated that areas of low population density had more alcohol-related motor vehicle crashes than expected (P < 0.05). There was no significant association between alcohol outlets and alcohol-related motor vehicle crashes using distance analyses, logistic regression, and Chi-square. Differences in environmental or behavioral factors characteristic of areas of low population density may be responsible for the higher proportion of alcohol-related crashes occurring in these areas. PMID- 15350892 TI - Pathogenesis of age-related macular degeneration. PMID- 15350893 TI - The role of NOS3 in stem cell mobilization. PMID- 15350894 TI - Stem cell aging and autoimmunity in rheumatoid arthritis. PMID- 15350895 TI - CIDE-A, a novel link between brown adipose tissue and obesity. PMID- 15350896 TI - Seipin: a mysterious protein. PMID- 15350897 TI - CDKs: taking on a role as mediators of dopaminergic loss in Parkinson's disease. PMID- 15350898 TI - Cdk5 deregulation in the pathogenesis of Alzheimer's disease. PMID- 15350899 TI - CD46 in Neisseria pathogenesis. PMID- 15350900 TI - The von Hippel-Lindau tumour suppressor: a multi-faceted inhibitor of tumourigenesis. PMID- 15350901 TI - The spectroscopic study of hydrogen bonding in some 3,3'-derivatives of 2,2' bipyridyl. AB - Four 3,3'-derivatives of 2,2'-bipyridyl have been investigated by multinuclear NMR, IR and X-ray methods. In all cases the weak intramolecular hydrogen bonds between exocyclic nitrogen-containing substituent and pyridine-type ring nitrogen atom were found. In contrast to the previous results the nitrogen chemical shifts of pyridine ring atom do not provide valuable information about hydrogen bond strength. The presence of intramolecular hydrogen bonds were confirmed by nitrogen chemical shifts of exocyclic amino and acetamide groups, deuterium isotope effects in the solid state and IR measurements in both chloroform solution and the solid state. The X-ray structures obtained for asymmetric 3 amino-3'-methylamino and 3,3'-diacetamide derivatives confirmed conclusions made on the base of spectral results. PMID- 15350902 TI - Curve-fitting of Fourier manipulated spectra comprising apodization, smoothing, derivation and deconvolution. AB - We present a general method for curve-fitting Fourier manipulated spectra, comprising apodized, smoothed, derivatised and deconvoluted spectra. The analytical expressions of Fourier manipulated bands in the spectral domain, needed for the curve-fitting, are usually very complex or do not even exist; hence an accurate curve-fit of Fourier manipulated spectra becomes unfeasible. Our strategy is to construct both the model and their derivatives in the Fourier domain, where they have simple and general expressions, and then Fourier transform them back to the spectral domain. The first benefit of this approach is the accurate curve-fitting of Fourier deconvoluted spectra, a main step in the secondary structure estimation of proteins by FTIR spectroscopy. PMID- 15350903 TI - Liquid/air interfacial structure of alcohol-octyl hydroxamic acid mixtures: a study by sum-frequency spectroscopy. AB - The molecular structure of the liquid/air interfaces of 1-octanol, 1-decanol, n decane and the branched decyl alcohol EXXAL 10 has been studied by sum-frequency spectroscopy (SFS) in the C-H stretching vibrational region. The data suggest that the interfaces consist of ordered molecules with closely packed alkyl tails, in close to all-trans conformation with some gauche defects. The degree of surface ordering for the branched alcohol is much higher than for octanol and decanol. When octyl hydroxamic acid (OHA) is dissolved in 1-octanol it increases the gauche conformational defects in the interfacial chains, possibly due to mixing with the surface alcohol molecules and disrupting their ordering. In contrast, we suggest that when octyl hydroxamic acid is dissolved in EXXAL 10, the surface ordering of the alcohol chains does not change. We put forward the hypothesis that the appearance of new bands, belonging to the asymmetric methylene group vibrations and to the asymmetric methyl modes in the SF spectra of the mixture suggests that the surface OHA molecules are arranged with their hydrocarbon tails tilted very close to the interface. PMID- 15350904 TI - Vibrational frequencies and structural determination of 1,3-dichloro-1,3 diazetidine-2,4-dione. AB - The normal mode frequencies and corresponding vibrational assignments of 1,3 dichloro-1,3-diazetidine-2,4-dione are examined theoretically using the Gaussian 98 set of quantum chemistry codes. Each of the vibrational modes was assigned to one of six types of motion predicted by a group theoretical analysis (C=O stretch, N-C stretch, N-Cl stretch, N-C-N bend, N-Cl bend, and C=O bend) utilizing the C2h symmetry of the molecule. Uniform scaling factors was derived for each type of motion. Predicted infrared and Raman intensities are reported. PMID- 15350905 TI - Investigations of kinetics and mechanism of chloropinnoite in boric acid aqueous solution at 303 K by Raman spectroscopy. AB - Raman spectroscopy of dissolution and transformation of chloropinnoite in 4.5% (wt.%) boric acid aqueous solution at 303 K has been recorded. The Raman spectra of kinetics process have been obtained. The phase transformation product is 2MgO.3B2O3.15H2O (kurnakovite). The main polyborate anions and their interaction in aqueous solution have been proposed according to the Raman spectrum. Some assignments were tentatively given and the relations between the existing forms of polyborate anions and the crystallizing solid phases have been gained. A mechanism of dissolution and crystallization reactions and the formation condition of kurnakovite in Qinghai-Tibet plateau were proposed and discussed. PMID- 15350906 TI - Intramolecular charge transfer dual fluorescence of p-dimethylaminobenzoates. AB - A series of ester, p-N,N-dimethylaminobenzoates ((CH3)2NC6H4COOR, R=n-CnH2n+1, n=1,2,...8), were synthesized and their fluorescence spectra were recorded. Intramolecular charge transfer (CT) dual fluorescence was found in common organic solvent or in water with cetyltrimethylammonium bromide (CTAB) micelle. The fluorescence intensity ratio of CT band to normal band (ICT/ILE) was decreased dramatically with the increasing of carbon number of R group when the carbon number of R group were 1-3, but the dependence was not obvious when the carbon number of R group were 4-8. Oppositely, the ICT/ILE was increased with increasing of carbon number of R group in water with CTAB micelle. PMID- 15350907 TI - Polarised infrared and Raman studies of YCa4O(BO3)3 a non-linear optical single crystal. AB - YCa4O(BO3)3-(YCOB) is a non-linear optical (NLO) material grown by Czochralski technique. Polarised IR, ATR-IR, polarised Raman and optical transmission spectral measurements were made. A series of absorption bands have been observed with intensities depending on the functional groups of the crystals. The observed bands were assigned and discussed. PMID- 15350908 TI - Spectroscopic characterization of fluorescein- and tetramethylrhodamine-labeled oligonucleotides and their complexes with a DNA template. AB - We measured absorption and emission spectra, fluorescence quantum yield, anisotropy, fluorescence resonance energy transfer (FRET), and melting temperature to characterize fluorescein- and tetramethylrhodamine (TMR)-labeled oligonucleotides in solution and when hybridized to a common DNA template. Upon hybridization to the template, both the absorption and emission spectra of TMR labeled duplexes exhibited a shift with respect to those of labeled oligonucleotides, depending on the location of the TMR on the oligonucleotide. Measurements of quantum yield, anisotropy, and melting temperature indicated that TMR interacted with nucleotides within the duplexes in the order (T1>T5>T11, T16) that the oligonucleotide with TMR labeled at the 5' end (T1) is stronger than that labeled at position 5 from the 5' end (T5), which is also stronger than those labeled at the positions, 11 and 16, from the 5' end (T11, T16). In the case of the duplex formed between T1 and the template, fluorescence quenching was observed, which is attributed to the interaction between the dye molecule and guanosines located at the single-stranded portion of the template. A two-state model was suggested to describe the conformational states of TMR in the duplex. The melting temperatures of the four FRET complexes show the same pattern as those of TMR-labeled duplexes. We infer that the interactions between TMR and guanosine persist in the FRET complexes. This interaction may bring the donor and the acceptor molecules closely together, which could cause interaction between the two dye molecules shown in absorbance measurements of the FRET complexes. PMID- 15350909 TI - Vibrational frequencies and structural determination of diethynyldimethylsilane. AB - The normal mode frequencies and corresponding vibrational assignments of diethynyldimethylsilane are examined theoretically using the Gaussian 98 set of quantum chemistry codes. Each of the vibrational modes was assigned to one of nine types of motion predicted by a group theoretical analysis (Si-C stretch, C[triple bond]C stretch, C-H stretch, C[triple bond]C-H bend, Si-C[triple bond]C bend, C-Si-C bend, H-C-H bend, CH3 wag, and CH3 twist) utilizing the C3v symmetry of the molecule. A set of uniform scaling factors was derived for each type of motion. Predicted infrared and Raman intensities are reported. PMID- 15350910 TI - Vibrational spectroscopic studies of vinyltriethoxysilane sol-gel and its coating. AB - Infrared and Raman spectra of vinyltriethoxysilane (VTES), the VTES sol-gel, and the sol-gel coated aluminum have been collected. The assignments of the vibrational modes for the silane, the sol-gel, and the sol-gel films have been made based on the group frequencies and the spectral variation collected at different physical and chemical states of the sol-gel samples. Applying the sol gels onto the metal grids allowed the drying and the high temperature treatment of the samples for the collection of infrared transmission spectra. From the variation of the sol-gel and the sol-gel coated aluminum IR spectra with temperature, it was noticed that the samples partially decomposed when the temperature was higher than 100 degrees C. Electrochemical experiments have demonstrated that the anticorrosion property of the coated aluminum has significantly increased. The water and the hexadecane contact angle measurements showed that the surface modified metal had a much higher hydrophobic property than the untreated metal. PMID- 15350911 TI - Spectroscopic characterization of tetradentate macrocyclic ligand: its transition metal complexes. AB - Manganese(II), cobalt(II), nickel(II) and copper(II) complexes are synthesized with a novel tetradentate ligand viz. 1,3,9,11-tetraaza-4,8,12,16-tetraoxo 2,6,10,14-tetrathiacyclohexadecane (L) and characterized by the elemental analysis, molar conductance measurements, magnetic susceptibility measurements, electron impact mass, 1H NMR, IR, electronic and EPR spectral studies. The molar conductance measurements of the complexes in DMSO correspond to be nonelectrolytic nature for Mn(II), Co(II) and Cu(II) while 1:2 electrolytes for Ni(II) complexes. Thus these complexes may be formulated as [M(L)X2] and [Ni(L)]X2 (where M: Mn(II), Co(II), and Cu(II) and X = Cl- and NO3-). On the basis of IR, electronic and EPR spectral studies an octahedral geometry has been assigned for Mn(II) and Co(II) complexes, square-planar for Ni(II) whereas tetragonal for Cu(II) complexes. The ligand and its complexes were also evaluated against the growth of bacteria and pathogenic fungi in vitro. PMID- 15350912 TI - Gas phase spectroscopy of HNO3 in the region 2000-8500 cm(-1). AB - Spectra of gas phase HNO3 were collected in the region 2000-8500 cm(-1) using Fourier-transform infrared spectroscopy. This region is dominated by the nu1 O-H stretching mode but also contains many previously unreported combination bands and overtones. This work marks the first observation of Fermi resonance the 2nu1 O-H stretching overtone. Previously unobserved bands were assigned and integrated intensities were obtained. For bands already reported in the literature, comparisons of relative intensities are presented when possible. This work gives a brief discussion on the trends in overtone intensities and on mode mixing in HNO3 in relation to previous experimental and theoretical studies. PMID- 15350913 TI - Expeditious implementation of two new methods for analysing the pigment composition of photosynthetic specimens. AB - Two new methods for analysing the pigment composition of photosynthetic samples have been developed during the last few years. One, called the spectral reconstruction method, uses linear least-squares fitting, while the other, named the Gauss-peak spectra method, entails non-linear optimisation; each has some advantages over the other, but both use a large number of data points and surpass the traditional method which uses absorbance at a few wavelengths. In order to make the new methods transparent to experimentalists who are not well versed in statistical analysis, curve fitting and interpolation, simple procedures are described for implementing the methods with the aid of a spreadsheet. The problem of analysing a sample containing chlorophyll c, which is difficult to isolate in a form sufficiently pure to serve as an analytical standard, is also briefly addressed. PMID- 15350914 TI - Photothermal spectrometry for detection in miniaturized systems: relevant features, strategies and recent applications. AB - There is a growing interest in using miniaturized analytical devices because they allow to execute the different steps of an analytical process within very short times and with drastic reduction in the amounts of solvents, reagents and samples. As for capillary electrophoresis, these systems require detectors which are sensitive, versatile and adaptable to very small detection volumes. In this respect, photothermal spectrometry which is complementary to fluorescence seems to be a promising detection method. This review describes the basic principle of photothermal spectrometry along with the related methods based on colinear-beam or crossed-beam configuration of the pump and probe lasers. Two experimental set ups especially designed for microfluidic systems as well as for capillary electrophoresis are described. Their characteristics and key features are discussed and the main applications are outlined. PMID- 15350915 TI - A novel method for preparation of cobalt(II) and lead(II) carbonates. AB - Cobalt(II) carbonate, CoCO3.4H2O and lead(II) carbonate, PbCO3.2H2O were synthesis by a new simple method during the reaction of aqueous solutions of CoX2 (X=Cl-, NO3- and CH3COO-) and PbX2 (X=NO3- or CH3COO-), respectively, with urea at approximately 85 degrees C for 2 h. The infrared spectra of the reaction products clearly indicates the absence of the bands due to coordinated urea, but show the characteristic bands of ionic carbonate. A general mechanisms describing the formation of cobalt and lead carbonates are suggested. PMID- 15350916 TI - Spectroscopic studies on some azo compounds and their cobalt, copper and nickel complexes. AB - Synthesis and characterization of [o-carboxy phenylazo] moiety of barbituric acid, thiobarbituric acid, thiouracil, citrazinic acid, and disodium chromotropate and their complexes derived from cobalt (II), nickel (II), and copper (II) salts were done. The stereochemistry and the mode of bonding of the complexes were achieved based on elemental analysis, NMR, UV-Vis, IR and ESR. The dissociation constants of the free azo ligands are evaluated by spectrophotometric methods. PMID- 15350917 TI - Vibrational frequencies and structural determination of triethynylmethylgermane. AB - The normal mode frequencies and corresponding vibrational assignments of triethynylmethylgermane are examined theoretically using the Gaussian98 set of quantum chemistry codes. Each of the vibrational modes was assigned to one of nine types of motion predicted by a group theoretical analysis Ge-C stretch, C[triple bond]C stretch, C-H stretch, C[triple bond]C-H bend, Ge-C[triple bond]C bend, C-Ge-C bend, H-C-H bend, CH3 wag, and CH3 twist) utilizing the C3v symmetry of the molecule. Uniform scaling factors were derived for each type of motion. Predicted infrared and Raman intensities are reported. PMID- 15350918 TI - Spectroscopic characterization of copper(II) complexes of indoxyl N(4)-methyl thiosemicarbazone. AB - New copper(II) complexes of indoxyl thiosemicarbazone (ITSC) of general composition CuL2X2 (where L: ITSC; X: Cl-, NO3-, ClO4-, NCS-) have been synthesized and characterized by elemental analysis, molar conductance, magnetic susceptibility measurements and spectral (electronic, IR, EPR, 1H NMR, Mass) studies. Cyclic voltammetry measurements show quasi-reversible Cu2+/Cu1+ couple. Various physico-chemical techniques suggest a tetragonal structure for these copper(II) complexes. PMID- 15350919 TI - Spectrophotometric determination of certain cephalosporins in pure form and in pharmaceutical formulations. AB - A simple and reproducible spectrophotometeric method for the assay of cefotaxime sodium, cefuroxime sodium, and ceftriaxone disodium with metol-chromium(VI) reagent has been developed. The procedure is based on direct oxidation of metol by potassium dichromate in presence of drug in acidic medium and subsequent formation of ternary complex. Beer's law is obeyed in the range 0.2-28 microg ml( 1) at lambdamax 520 nm. For more accurate analysis, Ringbom optimum concentration range is found to be 0.8-26.5 microg ml(-1). The molar absorptivity and Sandell sensitivity were calculated. Six replicate analysis of solutions containing seven different concentrations of the examined drugs were carried out and gave a mean correlation coefficient < or =0.9996; the factors of the regression line equation for the three cephalosporins were calculated. The proposed method was applied to the determination of the examined drugs in pharmaceutical formulations and the results demonstrated that the method is equally accurate, precise, and reproducible as the official methods. PMID- 15350920 TI - Solid state imposed vibrational coupling between nu1 and nu3 modes of tetrahedral ions. AB - Raman spectra of the thiomolybdate ion in the compounds [Cu.NH4][MoS4] and X2[MoS4], where X=NH4+, Rb+ or Cs+, are reported and used to confirm the hypothesis presented in an earlier paper [J. Chem. Phys. 94 (1991) 5946], that there is solid state induced vibrational coupling between the nu1 and nu3 modes of tetrahedral ions. PMID- 15350921 TI - Structure investigation, spectral, thermal, X-ray and mass characterization of piroxicam and its metal complexes. AB - [M(H2L)2](A)2.yH2O (where H2L: neutral piroxicam (Pir), A: Cl- in case of Ni(II) or acetate anion in case of Cu(II) and Zn(II) ions and y=0-2.5) and [M(H2L)3](A)z.yH2O (A: SO4(2-) in case of Fe(II) ion (z=1) or Cl(-) in case of Fe(III) (z=3) and Co(II) ions (z=2) and y=1-4) chelates are prepared and characterized using elemental analyses, IR, magnetic and electronic reflectance measurements, mass spectra and thermal analyses. IR spectra reveal that Pir behaves a neutral bidentate ligand coordinated to the metal ions through the pyridyl-N and carbonyl-O of the amide moiety. The reflectance and magnetic moment measurements reveal that these chelates have tetrahedral, square planar and octahedral geometrical structures. Mass spectra and thermal analyses are also used to confirm the proposed formulae and the possible fragments resulted from fragmentation of Pir and its chelates. The thermal behaviour of the chelates (TGA and DTA) are discussed in detail and the thermal stability of the anhydrous chelates follow the order Ni(II) congruent with Cu(II) Fe(II) or = 10 years compared with normal weight men. In women there was a graded increase (p < 0.001) in the likelihood of numerous metabolic disorders when moving from the normal weight to overweight < 10 year to overweight > or = 10 year categories. CONCLUSIONS: The duration of overweight has a significant effect on obesity-related comorbidities in men and women. PMID- 15350960 TI - Putative glucosensing property in rat and human activated microglia. AB - Microglial cells involved in the pathogenesis of many neurodegenerative diseases acquire the features of cytotoxic and phagocytic cells in response to certain pathogens and inflammatory signals. K(ATP) channels are energy sensors of ATP availability that link the cell's metabolic state to its membrane excitability. In pancreatic beta cells, they promote glucose-dependent insulin secretion, and in neurones, hyperpolarization that protects against hypoxic damage. This study analyses activated microglia in an in vivo rat neurodegenerative model based on acute hippocampal glutamate receptor overactivation and in postmortem samples from patients with Alzheimer's disease. We demonstrate that in activated microglia the K(ATP) channel components SUR-1 or SUR-2 are present together with glucokinase. Our results indicate that, according to glucose availability, these channels may modify microglia membrane potential. The functional relevance of these channels is seen as a new mechanism modulating the effects of external signals on microglia. PMID- 15350961 TI - Impaired long-term spatial and recognition memory and enhanced CA1 hippocampal LTP in the dystrophin-deficient Dmd(mdx) mouse. AB - Duchenne muscular dystrophy (DMD) is associated with cognitive deficits that may result from dystrophin deficiency in neurons. However, in the dystrophin deficient Dmd(mdx) mouse model of DMD, the nature of the memory impairment is not well characterised and its biological substrate is uncertain. Here, we demonstrate that dystrophin deficiency in Dmd(mdx) mice impairs long-term, but not short-term, object recognition memory and impairs long-term spatial memory, but not acquisition, following massed training in the water maze. Furthermore, we show that the abnormal enhancement of CA1 hippocampal LTP in Dmd(mdx) mice is not restricted to short-lasting mechanisms, but also affects the maintenance phase of LTP of both synaptic efficacy and neuronal excitability. We conclude that dystrophin loss alters memory consolidation in both spatial and nonspatial learning tasks, at least in part due to altered synaptic plasticity mechanisms, and suggest that the severity of the deficits may depend on the nature of the training procedure. PMID- 15350962 TI - Effects of vascular endothelial growth factor (VEGF) on motor neuron degeneration. AB - Both in mice and humans, low expression levels of vascular endothelial growth factor (VEGF) are linked to adult-onset motor neuron disease or amyotrophic lateral sclerosis (ALS). The mechanism through which reduced VEGF levels result in this phenotype is unknown. We therefore examined the direct effects of VEGF on motor neurons and found VEGF to have a direct neurotrophic effect on motor neurons in vitro. Survival and vulnerability to excitotoxicity of motor neurons from VEGF(delta/delta) mice was however similar to that of motor neurons from non transgenic littermates. The VEGF concentration in the spinal cord of mutant (G93A) SOD1 mice was not different from that found in wild-type SOD1 overexpressing mice. Upregulation of VEGF in the spinal cord, by housing mutant (G93A) SOD1 mice in hypoxic conditions, did not affect their life span. Our results show that VEGF is a neurotrophic factor for motor neurons in vitro, and shortage of this neurotrophic factor may contribute to the motor neuron death observed in humans and animals with low VEGF expression levels. PMID- 15350963 TI - The pathobiology of moderate diffuse traumatic brain injury as identified using a new experimental model of injury in rats. AB - Experimental models of traumatic brain injury have been developed to replicate selected aspects of human head injury, such as contusion, concussion, and/or diffuse axonal injury. Although diffuse axonal injury is a major feature of clinical head injury, relatively few experimental models of diffuse traumatic brain injury (TBI) have been developed, particularly in smaller animals such as rodents. Here, we describe the pathophysiological consequences of moderate diffuse TBI in rats generated by a newly developed, highly controlled, and reproducible model. This model of TBI caused brain edema beginning 20 min after injury and peaking at 24 h post-trauma, as shown by wet weight/dry weight ratios and diffusion-weighted magnetic resonance imaging. Increased permeability of the blood-brain barrier was present up to 4 h post-injury as evaluated using Evans blue dye. Phosphorus magnetic resonance spectroscopy showed significant declines in brain-free magnesium concentration and reduced cytosolic phosphorylation potential at 4 h post-injury. Diffuse axonal damage was demonstrated using manganese-enhanced magnetic resonance imaging, and intracerebral injection of a fluorescent vital dye (Fluoro-Ruby) at 24-h and 7-day post-injury. Morphological evidence of apoptosis and caspase-3 activation were also found in the cerebral hemisphere and brainstem at 24 h after trauma. These results show that this model is capable of reproducing major biochemical and neurological changes of diffuse clinical TBI. PMID- 15350964 TI - Nitric oxide synthase and intermittent hypoxia-induced spatial learning deficits in the rat. AB - Intermittent hypoxia (IH) during sleep induces significant neurobehavioral deficits in the rat. Since nitric oxide (NO) has been implicated in ischemia reperfusion-related pathophysiological consequences, the temporal effects of IH (alternating 21% and 10% O(2) every 90 s) and sustained hypoxia (SH; 10% O(2)) during sleep for up to 14 days on the induction of nitric oxide synthase (NOS) isoforms in the brain were examined in the cortex of Sprague-Dawley rats. No significant changes of endothelial NOS (eNOS) and neuronal NOS (nNOS) occurred over time with either IH or SH. Similarly, inducible NOS (iNOS) was not affected by SH. However, increased expression and activity of iNOS were observed on days 1 and 3 of IH (P < 0.01 vs. control; n = 12/group) and were followed by a return to basal levels on days 7 and 14. Furthermore, IH-mediated neurobehavioral deficits in the water maze were significantly attenuated in iNOS knockout mice. We conclude that IH is associated with a time-dependent induction of iNOS and that the increased expression of iNOS may play a critical role in the early pathophysiological events leading to IH-mediated neurobehavioral deficits. PMID- 15350965 TI - Synergistic effects of dopamine and Zn2+ on the induction of PC12 cell death and dopamine depletion in the striatum: possible implication in the pathogenesis of Parkinson's disease. AB - The mechanism that underlies the progressive degeneration of the dopaminergic neurons in Parkinson's disease (PD) is not clear. The Zn(2+) level in the substantia nigra of Parkinson's patients is increased. However, it is unknown whether Zn(2+) has a role in the degeneration of dopaminergic neurons. This study identifies an interaction between dopamine and Zn(2+) that induces cell death. When PC12 cells were pretreated with Zn(2+) before dopamine treatment, dopamine and Zn(2+) synergistically increased cell death, while Zn(2+) and H(2)O(2) had only additive effects on cell death. The synergistic effect appeared to be caused by increased apoptosis rather than necrosis. The synergistic effect was specific for Zn(2+). The synergistic effect was inhibited by thiol antioxidants but was not significantly affected by calcium channel blockers. There is a similar synergistic effect when dopamine and Zn(2+) were coinfused into the striatum, resulting in striatal dopamine content depletion in vivo. Thus, both dopamine oxidation and Zn(2+) are possibly linked to the degeneration of dopaminergic neurons. PMID- 15350966 TI - Inflammation alters cation chloride cotransporter expression in sensory neurons. AB - Cation chloride cotransporters have been proposed to play a role in the modulation of neuronal responses to gamma-aminobutyric acid (GABA). In conditions of neuronal damage, where neuronal excitability is increased, the expression of the KCC2 transporter is decreased. This is also seen in spinal cord in models of neuropathic pain. We have investigated the expression of the Na-K-Cl, and K-Cl cotransporters NKCC1 and KCC2, in dorsal root ganglion (DRG) and spinal sensory neurons during arthritis, a condition in which neuronal excitability is also increased. NKCC1 was expressed in control DRG neurons, and its expression was decreased in arthritis. Both NKCC1 and KCC2 were expressed in sensory neurons in the spinal cord. In acute arthritis, both NKCC1 and KCC2 mRNA increased in superficial but not deep dorsal horn, and this was accompanied by an increase in protein expression. In chronic arthritis, NKCC1 expression remained raised, but KCC2 mRNA and protein expression returned to control levels. Altered KCC2 and NKCC1 expression in arthritis may contribute to the control of spinal cord excitability and may represent novel therapeutic targets in the treatment of inflammatory pain. PMID- 15350967 TI - Inducible form of nitric oxide synthase expression in rat cortical neuronal cells in vitro. AB - The inducible form of nitric oxide synthase (iNOS) is an essential element of the immune response, which is expressed primarily in microglial cells within the CNS. Exposure of rat cortical neuronal cells to the pro-inflammatory bacterial endotoxin lipopolysaccharide (LPS) resulted in a significant increase in the expression of the cellular iNOS protein expression and NO generation (which serves as an indirect measure of NOS catalytic activity). These effects were potentiated by costimulation with interferon-gamma (IFNgamma) and the increase in NO generation was abolished by the iNOS selective inhibitor 1400W, although this did not attenuate the toxin-induced increase in the enzyme expression. As the cortex is one of the principal areas to be targeted in Alzheimer's disease (AD), the present findings may help to further our understanding of the biochemical events associated with the neurodegenerative process. PMID- 15350968 TI - Adenosine and glutamate extracellular concentrations and mitogen-activated protein kinases in the striatum of Huntington transgenic mice. Selective antagonism of adenosine A2A receptors reduces transmitter outflow. AB - The basal ganglia and deep layers of cerebral cortex neurodegeneration typically characterize the postmortem brain of Huntington disease (HD) patients. In this study, we employed 10- to 11-week-old transgenic HD mice (R6/2 line), in which the striatal adenosine extracellular levels, measured using the microdialysis technique, are significantly increased in comparison to wild-type mice. An increase in striatal adenosine is probably a precocious index of mitochondrial dysfunction that is described in both the postmortem brain of HD patients and transgenic mice striatal cells. The adenosine increase is matched by activation of the p38 mitogen-activated protein kinase (MAPK) in the striatal neurons of R6/2 mouse but not in the cortex. This result indicates that p38 MAPK is a correlate of striatal damage and suggests a role for p38 in the striatal neuron suffering and apoptosis described in this disease. The selective adenosine A(2A) receptor antagonist SCH 58261, administered through microdialysis fiber into the striatum, significantly decreases the outflow of glutamate in R6/2 mice. Antagonism of adenosine A(2A) receptors might be regarded as potentially useful in the treatment of this disease to control striatal excitotoxicity. PMID- 15350969 TI - Neonatal mice lacking functional Fas death receptors are resistant to hypoxic ischemic brain injury. AB - Neonatal hypoxia-ischemia (HI) upregulates Fas death receptor expression in the brain, and alterations in expression and activity of Fas signaling intermediates occur in neonatal brain injury. B6.MRL-Tnfrsf6(lpr) mice lacking functional Fas death receptors are protected from HI brain damage in cortex, striatum, and thalamus compared to wild-type mice. Expression of Fas death receptor and active caspases increase in the cortex after HI. In wild-type mice, the hippocampus is most severely injured, and the hippocampus is the only region not protected in the B6.MRL-Tnfrsf6(lpr) mice. The selective vulnerability of the hippocampus to injury correlates with (1) lower basal expression of [Fas-associated death-domain like IL-1beta-converting enzyme]-inhibitory protein (FLIP), (2) increased degradation of spectrin to its 145 or 150 kDa breakdown product, and (3) a higher percentage of non-apoptotic cell death following neonatal HI. We conclude that Fas signaling via both extrinsic and intrinsic caspase cascades causes brain injury following neonatal HI in a region-dependent manner. Basal levels of endogenous decoy proteins may modulate the response to Fas death receptor signaling and provide a novel approach to understanding mechanisms of neonatal brain injury. PMID- 15350970 TI - Neutral sphingomyelinase activation in endothelial and glial cell death induced by amyloid beta-peptide. AB - We have explored the molecular mechanism underlying amyloid beta-peptide (Abeta) mediated cytotoxicity in vitro. Exposure of murine cerebral endothelial cells (CECs) or C6 glioma cells to Abeta25-35 resulted in dose-dependent cell death. Ceramide is a pro-apoptotic lipid mediator. Forced elevation of cellular ceramide levels, either by application of an exogenous C2 ceramide analogue or bacterial sphingomyelinase that induces endogenous ceramide release from sphingomyelin, mimicked Abeta25-35 cytotoxicity in both CECs and C6 glioma cells. Abeta25-35 induced synthesis of ceramide was selectively mediated by activation of neutral sphingomyelinase (nSMase), but not acidic sphingomyelinase (aSMase) or ceramide synthase. Both 3-O-Me-SM and N-acetyl-L-cysteine, the selective and nonselective pharmacological inhibitors of nSMase, respectively, suppressed nSMase activation, ceramide production, and cytotoxic action induced by Abeta25-35 in CECs. Furthermore, genetic knockdown of nSMase by an antisense strategy rendered C6 glioma cells specifically resistant to Abeta25-35 cytotoxicity without affecting their vulnerability to serum deprivation. Together, nSMase activation with subsequent ceramide production may contribute, at least partially, to Abeta25-35 cytotoxicity in cell types with cerebral endothelial and glial lineage. PMID- 15350971 TI - Neuronal RNA oxidation is a prominent feature of familial Alzheimer's disease. AB - An in situ approach was used to identify the oxidized RNA nucleoside 8 hydroxyguanosine (8OHG) in the frontal cortex of familial Alzheimer's disease (FAD) with a mutation in presenilin-1 (PS-1) or amyloid beta protein precursor (AbetaPP) gene (n = 13, age 47-81 years). Neurons with marked 8OHG immunoreaction in the cytoplasm were widely distributed in the superior/middle frontal gyrus of FAD. Relative intensity measurements of neuronal 8OHG immunoreactivity showed that there was a significant increase in FAD compared with controls (n = 15, age 59-81 years), while there was no difference in relative 8OHG between the PS-1 and the AbetaPP FAD. Interestingly, a presymptomatic case carrying a PS-1 mutation showed a considerable level of relative 8OHG, and the increased levels of neuronal 8OHG in FAD were more prominent in cases with a lower percentage area of Abeta42 burden. These results suggest that oxidative stress is an early event involved in the pathological cascade of FAD. PMID- 15350972 TI - Human single chain Fv antibodies and a complementarity determining region-derived peptide binding to amyloid-beta 1-42. AB - A library of phage-displayed human single-chain Fv (scFv) antibodies was selected against the human amyloid-beta peptide (Abeta42). Two new anti-Abeta42 phage displayed scFvs antibodies were obtained, and the sequences of their V(H) and Vkappa genes were analyzed. A synthetic peptide based on the sequence of Ig heavy chain (V(H)) complementarity-determining region (HCDR3) of the clone with the highest recognition signal was generated and determined to bind to Abeta42 in ELISA. Furthermore, we showed for the first time that an HCDR3-based peptide had neuroprotective potential against Abeta42 neurotoxicity in rat cultured hippocampal neurons. Our results suggest that not only scFvs recognizing Abeta42 but also synthetic peptides based on the V(H) CDR3 sequences of these antibodies may be novel potential candidates for small molecule-based Alzheimer's disease (AD) therapy. PMID- 15350973 TI - CUPpling calcium to lysosomal biogenesis. AB - Transport from late endosomes to lysosomes results in the formation of an endosome-lysosome hybrid organelle from which late endosomes and lysosomes must be re-formed. Recent studies indicate that the transient receptor potential (TRP) related channel mucolipin-1 (the loss of which causes mucolipidosis type IV) and its Caenorhabditis elegans orthologue CUP-5 might control the process of lysosome re-formation by regulating calcium flux. PMID- 15350974 TI - Checkpoints in ER-associated degradation: excuse me, which way to the proteasome? AB - The failure of secreted proteins to fold results in their retrotranslocation from the endoplasmic reticulum (ER) and degradation by the proteasome in a process called "ER-associated degradation" (ERAD). Two recent studies indicate that ERAD substrates are targeted to different pathways depending on the topology of the substrate and the subcellular location of the misfolded domain. PMID- 15350975 TI - Modulated receptor interactions in bacterial transmembrane signaling. AB - Bacteria can detect and respond to a remarkably diverse set of environmental conditions. This ability enables motile species to integrate stimuli, to compare current surroundings with those of the recent past, and to adjust swimming behavior to move up gradients of attractants and avoid repellents. Many of the molecular details involved in the bacterial chemotaxis system have been elucidated. Several models have been proposed recently to explain how cells process external information through a patch of highly interactive transmembrane receptors and transduce this information to other components in the cytoplasm that, in turn, function to regulate motility. PMID- 15350976 TI - Short-range intracellular trafficking of small molecules across endoplasmic reticulum junctions. AB - Intracellular trafficking is not mediated exclusively by vesicles. Additional, non-vesicular mechanisms transport material, in particular small molecules such as lipids and Ca(2+) ions, from one organelle to another. This transport occurs at narrow cytoplasmic gaps called membrane contact sites (MCSs), at which two organelles come into close apposition. Despite the conservation of these structures throughout evolution, little is known about this transport, largely because of a lack of knowledge of almost all molecular components of MCSs. Recently, this situation has started to change because the structural proteins that bridge an MCS are now known in a single case, and proteins implicated in lipid trafficking have been localized to MCSs. In the light of these advances, I hypothesize that the endoplasmic reticulum has a central role in the trafficking of lipids and ions by forming a network of MCSs with most other intracellular organelles. PMID- 15350977 TI - Surfing, regulating and capturing: are all microtubule-tip-tracking proteins created equal? AB - A diverse group of microtubule-binding proteins has been linked through live-cell imaging of green fluorescent protein (GFP) fusion proteins. These proteins share the ability to associate with the plus ends of elongating microtubules and track with these tips as the microtubules grow, in a process known as "tip tracking". Several models have been proposed to explain the significance of this activity, including roles in delivering proteins to the cell periphery and in modulating microtubule dynamics. However, the recent observation that some of the tip trackers colocalize on structures undergoing search-capture suggests that tip tracking could be a fundamental aspect of the search-capture process. Focusing on the shared ability of these proteins to undergo tip tracking, this article is intended to place the search-capture model in the context of other proposed functions and to stimulate discussion in this area. PMID- 15350978 TI - Potentials and pitfalls of fluorescent quantum dots for biological imaging. AB - Fluorescent semiconductor nanocrystals, known as quantum dots (QDs), have several unique optical and chemical features. These features make them desirable fluorescent tags for cell and developmental biological applications that require long-term, multi-target and highly sensitive imaging. The improved synthesis of water-stable QDs, the development of approaches to label cells efficiently with QDs, and improvements in conjugating QDs to specific biomolecules have triggered the recent explosion in their use in biological imaging. Although there have been many successes in using QDs for biological applications, limitations remain that must be overcome before these powerful tools can be used routinely by biologists. PMID- 15350979 TI - Importin alpha: a multipurpose nuclear-transport receptor. AB - The importin alpha/beta heterodimer targets hundreds of proteins to the nuclear pore complex (NPC) and facilitates their translocation across the nuclear envelope. Importin alpha binds to classical nuclear localization signal (cNLS) containing proteins and links them to importin beta, the karyopherin that ferries the ternary complex through the NPC. A second karyopherin, the exportin CAS, recycles importin alpha back to the cytoplasm. In this article, we discuss control mechanisms that importin alpha exerts over the assembly and disassembly of the ternary complex and we describe how new groups of importin alpha genes arose during the evolution of metazoan animals to function in development and differentiation. We also describe activities of importin alpha that seem to be distinct from its housekeeping functions in nuclear transport. PMID- 15350980 TI - Peroxisome turnover by micropexophagy: an autophagy-related process. AB - Many organisms stringently regulate the number, volume and enzymatic content of peroxisomes (and other organelles). Understanding this regulation requires knowledge of how organelles are assembled and selectively destroyed in response to metabolic cues. In the past decade, considerable progress has been achieved in the elucidation of the roles of genes involved in peroxisome biogenesis, half of which are affected in human peroxisomal disorders. The recent discovery of intermediates and genes in peroxisome turnover by selective autophagy-related processes (pexophagy) opens the door to understanding peroxisome turnover and homeostasis. In this article, we summarize advances in the characterization of genes that are necessary for the transport and delivery of selective and nonselective cargoes to the lysosome or vacuole by autophagy-related processes, with emphasis on peroxisome turnover by micropexophagy. PMID- 15350981 TI - Neurological mechanisms of green tea polyphenols in Alzheimer's and Parkinson's diseases. AB - Tea consumption is varying its status from a mere ancient beverage and a lifestyle habit, to a nutrient endowed with possible prospective neurobiological pharmacological actions beneficial to human health. Accumulating evidence suggest that oxidative stress resulting in reactive oxygen species generation and inflammation play a pivotal role in neurodegenerative diseases, supporting the implementation of radical scavengers, transition metal (e.g., iron and copper) chelators, and nonvitamin natural antioxidant polyphenols in the clinic. These observations are in line with the current view that polyphenolic dietary supplementation may have an impact on cognitive deficits in individuals of advanced age. As a consequence, green tea polyphenols are now being considered as therapeutic agents in well controlled epidemiological studies, aimed to alter brain aging processes and to serve as possible neuroprotective agents in progressive neurodegenerative disorders such as Parkinson's and Alzheimer's diseases. In particular, literature on the putative novel neuroprotective mechanism of the major green tea polyphenol, (-)-epigallocatechin-3-gallate, are examined and discussed in this review. PMID- 15350982 TI - A multinational study of alpha-lactalbumin concentrations in human milk. AB - Alpha-lactalbumin, a 14-kD protein, plays a central biochemical role in the mammary gland as the regulatory subunit of lactose synthase, and also plays a nutritional role for the rapidly growing neonate as the protein in highest concentration in human milk. The current study was undertaken to better characterize alpha-lactalbumin concentrations in human milk from a variety of countries. Mature human milk (lactation duration > or =1 month) was collected from at least 50 women from nine different countries on five continents. Alpha lactalbumin concentration was determined by HPLC. The mean +/- SD for 452 samples was 2.44 +/- 0.64 g/L. The mean value of the samples from the United States was significantly higher than that from any other country, and the mean in Mexico was significantly lower than that from every country except China and Canada. Alpha lactalbumin concentration decreased with increasing duration of lactation and was positively correlated with total nitrogen. On average, alpha-lactalbumin contributed 16% of the total nitrogen content of human milk and consequently an important part of the amino acid content. PMID- 15350983 TI - Effect of the H, K-ATPase inhibitor, esomeprazole magnesium, on gut total antioxidant capacity in mice. AB - Antioxidant depletion is believed to be a mechanism involved in the pathophysiology of several upper gastrointestinal disorders, and H, K-ATPase inhibitors can alter free radical production by neutrophils. We hypothesized that the H, K-ATPase inhibitor esomeprazole magnesium would decrease gut free radical production with a concomitant increase in gut total antioxidant capacity. A/J mice (n = 10/group) received either vehicle (control) or one of three concentrations of esomeprazole magnesium in vehicle by once-daily gavage for 10 days. Using tissue extracts from stomach and colon, total antioxidant capacity, lipid peroxide levels, and constitutive Cu/Zn-superoxide dismutase were measured using validated assays. There was a dose-related increase in total antioxidant capacity (analysis of variance, P < 0.001) in stomach, but there was no change in the colon. In the assessment of free radical production, there was a trend toward decreased lipid peroxide levels in stomach from mice receiving esomeprazole. In stomach, Cu/Zn-superoxide dismutase activity was increased (ANOVA: p=.03) in mice receiving esomeprazole. In conclusion, gastric total antioxidant capacity and Cu/Zn-superoxide dismutase activity are increased by esomeprazole, and these changes may result in part from decreased free radical production. The present results support the notion that the pharmacological effects of this agent on upper intestinal tissue are more complex than previously thought, and appear to involve both enzymatic and nonenzymatic tissue antioxidants. PMID- 15350984 TI - The influence of beer with different antioxidant potential on plasma lipids, plasma antioxidant capacity, and bile excretion of rats fed cholesterol containing and cholesterol-free diets. AB - The aim of this investigation was to assess the influence of beers with different antioxidant potentials on plasma lipid metabolism, plasma antioxidant capacity, and bile excretion of rats fed cholesterol-containing and cholesterol-free diets. Four types of beers were investigated in vitro. Two of them (designated as BeerHigh and BeerLow) with the highest and lowest antioxidant potentials (34.5% and 21.4% and 2.07 mmol/L and 1.65 mmol/L according to beta-carotene assay and Trolox equivalent antioxidant coefficient, respectively), were chosen for the experiment on rats. A total of 60 male Wistar rats were divided into 6 dietary groups of 10 rats each; the groups were designated as Control, BeerA, BeerB, Chol, Chol/BeerA, and Chol/BeerB. The rats in the Control group were fed a basal diet (BD) only, which included wheat starch, casein, soybean oil, vitamin, and mineral mixtures. To the BD of the other five groups were added the following: BeerHigh (BeerA), BeerLow (BeerB), 1% of cholesterol (Chol), 1% of cholesterol and BeerHigh (Chol/BeerA), and 1% of cholesterol and BeerLow (Chol/BeerB). After 4 weeks of feeding, diets supplemented with BeerHigh and, to a lesser degree, with BeerLow (Chol/BeerA and Chol/BeerB groups) hindered a rise in plasma lipids and a decrease in plasma antioxidant capacity, and increased the bile excretion indices. Supplementation with BeerHigh and, to a lesser degree, with BeerLow in rats fed cholesterol-free diets increased their plasma antioxidant capacity. No significant changes in the plasma lipid levels, antioxidant capacity, and bile excretion indices were observed in the Control group. In conclusion, beer was found to have a positively effect on plasma lipid profile and plasma antioxidant capacity, and to increase the bile excretion indices in rats fed cholesterol containing diets. The degree of this positive influence is directly connected to the contents of the bioactive components and the related antioxidant potential of beer. It is suggested that to achieve the best results, beer with the highest antioxidant potential must be consumed. PMID- 15350985 TI - Biochemical responses of healthy subjects during dietary supplementation with L arginine. AB - Dietary supplements of L-arginine, a substrate for nitric oxide synthases, may promote formation of nitric oxide and thus may be of clinical benefit. However, the optimal level of L-arginine supplementation is unclear. The objective of this study was to evaluate the response of healthy individuals to increasing doses of L-arginine (as free acid). Twelve healthy subjects were recruited and instructed to take L-arginine for 1-week periods at daily doses of 3, 9, 21, and 30 g. At baseline and at the end of each week, 24-hour urine and fasting blood samples were collected, and weight, diastolic blood pressure, and systolic blood pressure were recorded. Samples were analyzed for L-arginine, L-citrulline, glycine, L lysine, L-ornithine, asymmetric dimethy L-arginine, symmetric dimethy L-arginine, glucose, insulin (serum), creatinine, cGMP (urine), and total nitrates (serum and urine). Ten subjects reported adverse side effects at initial L-arginine doses of 21 g/day (five subjects) or 30 g/day (five subjects), respectively. Blood pressure and weight did not change during the supplementation period. Of the individual biochemical measures, only L-arginine, glycine, and L-ornithine concentrations changed significantly. The mean concentration of L-arginine reached a peak during supplementation at 9 g/d; however, individuals differed markedly in their response. Availability of L-arginine, relative to that of asymmetric dimethy L-arginine, increased significantly at both 9 g/day and 21 g/day. Mean values indicate that supplementation with 9 g/day of L-arginine, a dose associated with minimal adverse side effects, is sufficient to increase circulating L-arginine concentrations. However, subjects varied widely in their responses, indicating that L-arginine supplementation needs to be tailored to individuals. PMID- 15350986 TI - Decreased aortic early atherosclerosis and associated risk factors in hypercholesterolemic hamsters fed a high- or mid-oleic acid oil compared to a high-linoleic acid oil. AB - Currently, diets higher in polyunsaturated fat are believed to lower blood cholesterol concentrations, and thus reduce atherosclerosis, greater than diets containing high amounts of saturated or possibly even monounsaturated fat. The present study was designed to investigate the effect of diets containing mid- or high-linoleic oil versus the typical high-linoleic sunflower oil on LDL oxidation and the development of early atherosclerosis in a hypercholesterolemic hamster model. Animals were fed a hypercholesterolemic diet containing 10% mid-oleic sunflower oil, high-oleic olive oil, or high-linoleic sunflower oil (wt/wt) plus 0.4% cholesterol (wt/wt) for 10 weeks. After 10 weeks of dietary treatment, only the animals fed the mid-oleic sunflower oil had significant reductions in plasma LDL-C levels (-17%) compared to the high-linoleic sunflower oil group. The high oleic olive oil-fed hamsters had significantly higher plasma triglyceride levels (+41%) compared to the high-linoleic sunflower oil-fed hamsters. The tocopherol levels in plasma LDL were significantly higher in hamsters fed the mid-oleic sunflower oil (+77%) compared to hamsters fed either the high-linoleic sunflower or high-oleic olive oil. Measurements of LDL oxidation parameters, indicated that hamsters fed the mid-oleic sunflower oil and high-oleic olive oil diets had significantly longer lag phase (+66% and +145%, respectively) and significantly lower propagation rates (-26% and -44%, respectively) and conjugated dienes formed (-17% and -25%, respectively) compared to the hamsters fed the high linoleic sunflower oil. Relative to the high-linoleic sunflower oil, aortic cholesterol ester was reduced by -14% and -34% in the mid-oleic sunflower oil and high-oleic olive oil groups, respectively, with the latter reaching statistical significance. Although there were no significant associations between plasma lipids and lipoprotein cholesterol with aortic total cholesterol and cholesterol esters for any of the groups, the lag phase of conjugated diene formation was inversely associated with both aortic total and esterified cholesterol in the high-oleic olive oil-fed hamsters (r = -0.69, P < 0.05). The present study suggests that mid-oleic sunflower oil reduces risk factors such as lipoprotein cholesterol and oxidative stress associated with early atherosclerosis greater than the typical high-linoleic sunflower oil in hypercholesterolemic hamsters. The high-oleic olive oil not only significantly reduced oxidative stress but also reduced aortic cholesterol ester, a hallmark of early aortic atherosclerosis greater than the typical high-linoleic sunflower oil. PMID- 15350987 TI - Dietary selenium supplementation prolongs pentobarbital induced hypnosis. AB - The present studies characterized the influence of dietary selenium (Na2SeO3) on the duration of pentobarbital (PB) induced hypnosis (sleep) in the rat. Rats were fed semipurified diets varying from 0.01 to 2.0 mg Se/kg for up to 4 weeks. Consumption of diets containing 1.0 and 2.0 mg Se/kg significantly prolonged PB induced hypnosis. Hepatic selenium, but not hepatic glutathione peroxidase activity, correlated with the length of PB induced hypnosis. The prolongation of hypnosis caused by diets containing 1.0 mg Se/kg was substantially reduced or eliminated by repeated exposure to PB. Although single exposure to increasing quantities of PB (60-100 mg/kg body weight) led to a progressive increase in sleep duration, the proportional increase caused by supplemental selenium (2.0 vs 0.1 microg Se/g) remained relatively constant (approximately 25%). Increasing maturity was inversely related to the duration of PB induced hypnosis, regardless of dietary selenium provided. Consumption of the 2.0 mg Se/kg diet prolonged PB induced hypnosis to a greater degree in immature than in mature rats (P < 0.05). Consumption of the selenium enriched diet (2 microg Se/g) resulted in an increase in cytochrome 2B, but had no effect on cytochrome 1A compared to controls (0.1 microg Se/g). Pretreatment of rats with P450 enzymes activators (i.e., PB, Aroclor 1254, or 3-methylcholanthrene) shortened the duration of PB induced sleep and masked the effects of dietary selenium. The current studies document that dietary selenium can influence the response to pentobarbital induced hypnosis and likely relates to changes in drug detoxification enzymes. PMID- 15350988 TI - Methylenetetrahydrofolate reductase 677C-->T polymorphism affects DNA methylation in response to controlled folate intake in young women. AB - DNA methylation is critical for normal genomic structure and function and is dependent on adequate folate status. A polymorphism (677C-->T) in a key folate enzyme, methylenetetrahydrofolate reductase (MTHFR), may impair DNA methylation when folate intake is inadequate and may increase the risk of reproductive abnormalities. The present study was designed to evaluate the effect of the MTHFR 677C-->T polymorphism on changes in global DNA methylation in young women consuming a low folate diet followed by repletion with the current Recommended Dietary Allowance (RDA). Women (age 20-30 years) with the TT (variant; n = 19) or CC (n = 22) genotype for the MTHFR 677C-->T polymorphism participated in a folate depletion-repletion study (7 weeks, 115 microg DFE/day; 7 weeks, 400 microg DFE/day). DNA methylation was measured at baseline, week 7, and week 14 using a [3H]methyl acceptance assay and a novel liquid chromatography tandem mass spectrometry assay of the DNA bases methylcytosine and cytosine. [3H]Methyl group acceptance tended to increase (P = 0.08) during depletion in all subjects, indicative of a decrease in global DNA methylation. During repletion, the raw change and the percent change in the methylcytosine/total cytosine ratio increased (P = 0.03 and P = 0.04, respectively) only in the subjects with the TT genotype. Moderate folate depletion in young women may cause a decrease in overall DNA methylation. The response to folate repletion suggests that following folate depletion women with the MTHFR 677 TT genotype have a greater increase in DNA methylation with folate repletion than women with the CC genotype. PMID- 15350989 TI - Structure-activity relationship of conjugated linoleic acid and its cognates in inhibiting heparin-releasable lipoprotein lipase and glycerol release from fully differentiated 3T3-L1 adipocytes. AB - Conjugated linoleic acid (CLA) reduces body fat in part by inhibiting the activity of heparin-releasable lipoprotein lipase (HR-LPL) activity in adipocytes, an effect that is induced by the trans-10,cis-12 CLA isomer. In this study we used a series of compounds that are structurally related to CLA (i.e., CLA cognates) to investigate the structural basis for this phenomenon. None of the 18:1 CLA cognates that were tested, nor trans-9,cis-12 18:2, cis-12-octadecen 10-ynoic acid (10y,cis-12) or 11-(2'-(n-pentyl)phenyl)-10-undecylenic acid (designated P-t10), exhibited any significant effect on HR-LPL activity. Among the CLA derivatives (alcohol, amide, and chloride) that were tested, only the alcohol form inhibited HR-LPL activity, although to a lesser extent than CLA itself. In addition, intracellular TG was reduced only by trans-10,cis-12 CLA and the alcohol form of CLA. Hence it appears that the trans-10,cis-12 conjugated double bond in conjunction with a carboxyl group at C-1 is required for inhibition of HR-LPL activity, and that an alcohol group can partially substitute for the carboxyl group. We also studied glycerol release from the cells, observing that this was enhanced by trans-10 18:1, trans-13 18:1, cis-12 18:1, cis-13 18:1, P-t10 but was reduced by cis-9 18:1, the alcohol and amide forms of CLA or 10y,cis-12. Accordingly the structural feature or features involved in regulating lipolysis appear to be more complex. Despite enhancing lipolysis in cultured 3T3-L1 adipocytes, trans-10 18:1 did not reduce body fat gain when fed to mice. PMID- 15350990 TI - [Pediatric practice by telephone: a difficult exercise]. PMID- 15350991 TI - [Pediatric telephone advice in the emergency department]. AB - OBJECTIVES: To describe the activity of telephone advice in a pediatric emergency department and assess the influencing factors to improve quality of care. METHODS: Descriptive study about all the anonymous telephone calls received on the direct line of the pediatric emergency room of Le Havre hospital, from 25 January to 25 July 2002, and all the advices given by a doctor or a nurse. RESULTS: The mean daily call frequency was 2.15 (0-12) with 586 calls during the 6 months period and the mean call duration was 3 min (1-20). Parents took telephone advices for: fever (27%), digestive troubles (22%), and trauma (14%). We found no difference concerning symptoms according to season. The rush hours were, on a bimodal graph, 0-1 am and 8-9 pm, paralleling the rush activity of consultation in pediatric emergency room. The heavy days for phone advices during the week were Tuesday and Wednesday. We found no correlation between heavy days of week and phone call duration. Thirty percent of cases did not need any advice because the asks were only an orientation advice. Advice to go to our emergency department was done in 11% of orientation advice. The call duration was significantly longer for: (1) calls including several symptoms or griefs, (2) calls given by a nurse; (3) calls taking place during hours of lowest activity in the emergency room (between 6 and 12 am), (4) calls including advice about medication or diet; (5) calls for counselling an orientation other than medical establishment. CONCLUSION: Our assessment of telephone call advices represented the first step to improve the quality of answer to families. Next step will be written protocols to answer more adequately to main griefs and symptoms that lead families to search for phone counselling. PMID- 15350992 TI - [Sickle cell disease pain management following the World Health Organization's protocol]. AB - Twenty-four percent of the Gabonese population has sickle cell trait, and 1-3% has sickle cell disease. Patients' management must follow well-defined protocols that take into account the debilitating effect of severe pain episodes. OBJECTIVES: To evaluate the three-step analgesic ladder for control of pain for sickle cell disease pain crises. METHODOLOGY: This is a prospective and descriptive study based on surveys filled between February 2000 and March 2001. Surveys were restricted to sickle cell disease patients having developed pain crises before and during their stay at the hospital. The assessment of the pain was based on the DEGR and EVA scales. The treatment followed the World Health Organisation's analgesic ladder. The criteria used for estimate pain severity and sedation duration were measured before, 2 and 24 h after treatment administration began (H0, H2 and H24) to evaluate its effectiveness. RESULTS: Sixty children were included. All the social layers of the population were represented. The school level was variable. Fifty-six percent of the patients suffered their first pain crisis before they were 1 year old. Thirty-one percent had annual crises and 20% monthly crises. Pain was located in joints in 35% of the cases, and in joints and bones in 15% of the cases. Eighty-one percent of the pain crises were successfully treated in stage I. The passage to stage II occurred in the remaining cases (18.3% of the cases). CONCLUSION: The WHO's three-step analgesic ladder for control of pain is effective for the sickle cell disease pain episodes. Stage I drugs relieved the pain for 81.6% of the patients in a relatively short time (2-16 h). It was not possible to go to stage III. PMID- 15350993 TI - [Hepatic abscesses in childhood: retrospective study about 33 cases observed in New-Caledonia between 1985 and 2003]. AB - Hepatic abscesses in childhood are rarely observed in Europe. The aim of this word was to study how to diagnose and how to treat an hepatic abscess. METHODS: Between 1985 and 2003, we recensed retrospectively 33 cases of hepatic abscesses hospitalised in the paediatric unit of Noumea. RESULTS: Children were mainly melanesians (79%), 7 years old on average, having abdominal pains, a clinical and biological infectious syndrome, and abscesses images on ultrasonography or computed tomography. The identified micro-organisms included Entamoeba histolytica in 30% (10 cases); Staphylococcus aureus in 15% (five cases), Staphylococcus coagulase negative in 6% (two cases), Streptococcus D in 3% (one case); Bartonella henselae in 9% (three cases); ascaris in 6% (two cases); Mycobacterium tuberculosis in 6% (two cases). In eight cases no bacteria was identified (24%) but the good evolution after antibiotics and the negative amoebic serology looked like pyogenic abscesses. Two abscesses were aspirated, two were drained, one child had a surgical intervention. There was no death. Following a mean duration of 1 month for antibiotics treatment, outcome was always favourable. CONCLUSION: Diagnosis of hepatic abscess can be difficult. Ultrasonography shows the abscess but not the causal agent. The amoebic serology is sensible, consequently, its negativity leads to evoke a pyogenic agent. Early antibiotic treatment against pyogenic, anaerobic bacteria, and Entamoeba histolytica is required. Hepatic abscesses in ascaridiosis, tuberculosis and cat scratch disease are less frequently encountered. If diagnosis remains doubtful or clinical evolution worsens, or if abscess volume increases, a percutaneous aspiration or drainage is needed. PMID- 15350994 TI - [Practice for insulin infusion in preterm infants]. AB - Transient neonatal hyperglycemia is commonly observed during the first week of life in the preterm infants less than 30 weeks of gestational age. Continuous insulin infusion is an effective treatment in this situation. OBJECTIVE: To ascertain how insulin is administered in different french neonatal intensive care units. MATERIAL AND METHODS: We surveyed 49 neonatal intensive care units with a questionnaire. Response rate was 77.5% (38/49). RESULTS: Thirty four of 38 neonatal intensive care units reported the use of insulin infusions in this setting. Glucose level indicating insulin therapy and the initial insulin doses were quite variable according to the different units (respectively 7-16.5 mmol/l and 0.01-0.1 U/kg/h). A range of minimal insulin concentrations was used (0.01 0.1 U/ml), 57% utilizing concentration between 0.05 and 0.2 U/ml. Flow rates below 0.3 ml/h were used at time by 76%. Albumin was rarely added. Fifty seven percent of the neonatal intensive care units took counter-measures such as preconditioning and flushing the tubing to control insulin loss due to adsorption. The counter-measures were differently applied. Despite these measures, hyperglycemia and insulin resistance were frequently observed (respectively 30% and 47%). The different practices are discussed according to the literature. CONCLUSION: In order to deliver insulin reliably, we suggest an insulin delivery method for the preterm infants. PMID- 15350995 TI - [Diffusing capacity for carbon monoxide (T(LCO)) and oxygen saturation during exercise in patients with cystic fibrosis]. AB - OBJECTIVES: To estimate the value of diffusing capacity for carbon monoxide (T(LCO)) in patients with cystic fibrosis and to evaluate its ability to predict arterial desaturation during exercise. METHOD: Fourty-four patients (9-30 years) with cystic fibrosis performed pulmonary function tests with measure of T(LCO) and a bicycle incremental exercise test. They represent a wide variation in disease severity: mean Shwachman score: 77.8 (range: 40-100), mean FEV1%: 72.8 (range: 17-131). This study investigated the relationship between T(LCO), lung volumes and exercise data. RESULTS: T(LCO) remained normal for a long time in patients with cystic fibrosis: 82% of them show a normal T(LCO) (mean value: 91.3% of predicted). T(LCO) was significantly correlated with FEV(1), residual volume, maximal work load and maximum oxygen uptake. A fall in arterial oxygen saturation was uncommon in our study (five patients) and not significantly correlated with T(LCO). CONCLUSIONS: T(LCO) is a good criter of severity of cystic fibrosis but remains unreliable to predict values above which physical activity is safe, without arterial desaturation. Exercise tests should be proposed in order to evaluate exercise adaptation of each patient and determine which factor limits maximal performance. PMID- 15350996 TI - [Spontaneous gastric perforation in a neonate. A case report]. AB - Gastric perforation in the neonate is rare. The authors report a 3-day-old male infant, born through a normal delivery after an uncomplicated pregnancy, who presented with a sudden gross distention of the abdomen followed by respiratory distress. Abdominal X-ray examination showed a massive pneumoperitoneum. After a brief resuscitation, an isolated perforation of the stomach was discovered on laparotomy and was sutured. Outcome of spontaneous gastric perforation in newborns is favourable provided the disease is early recognised and treated. PMID- 15350997 TI - [Acute bartholinitis caused by Pseudomonas aeruginosa in an 18-month-old infant]. AB - Acute bartholinitis is a disease usually seen in women in the period of genital activity. Its occurence in a prepubertal child is an extremely rare event. CASE REPORT: We describe the case of a 18-month-old infant presenting a Bartholin's gland abces caused by Pseudomonas Aeruginosa with a resolutive evolution after antibiotherapy and surgical drainage. CONCLUSION: Diagnosis of Bartholinitis should be considered in any female infant with a labial enlargement. PMID- 15350998 TI - [Oxcarbazepine and DRESS syndrome: a paediatric cause of acute liver failure]. AB - Drug rash with eosinophilia and systemic symptoms (DRESS) syndrome, also called hypersensitivity reaction, is a severe idiosyncratic reaction to drugs, especially to anti-epileptic drugs. Clinical features associate cutaneous eruption, fever, multiple peripheral ganglions, and potentially life-threatening damage of one or more organs. DRESS syndrome is well described in adults treated with aromatic anti-epileptic drugs, such as phenytoin, phenobarbital, and carbamazepine, but also with other drugs. The new anti-epileptic drugs, such as oxcarbazepine also induce various cutaneous eruptions, but with less report of DRESS syndrome. In children, DRESS syndrome is rare and probably underdiagnosed. We report on the case of a 11-year-old girl hospitalised with an acute severe hepatitis revealing an oxcarbazepine-induced DRESS syndrome. PMID- 15350999 TI - [Isolated agenesia of pulmonary artery]. AB - Unilateral primitive agenesia of pulmonary artery is a rare congenital disorder. Experience of three hospital-based pulmonology pediatric units including eight children (three boys and five girls) with such anomaly is reported (11 years median follow-up, range 6 months to 10 years). Median age at diagnosis was 4 years. Clinical features included recurrent respiratory tract infections (75%), effort dyspnea (50%) and, curiously, asthmatic symptoms (75%). Diagnosis was evoked on chest X rays and confirmed by angiography in all cases. Agenesis of the right pulmonary artery was more frequent (five cases). Abnormal vascular ring was associated in two cases of agenesis of the left pulmonary artery. Pulmonary function tests were performed in six children, with bronchial obstruction in two. Regular clinical follow-up, up to adult age in four cases, showed no complication, such as hemoptysis or pulmonary hypertension in our patients. PMID- 15351000 TI - [Molecular mechanism of edema formation in nephrotic syndrome]. AB - Nephrotic edema are the clinical feature of isolated interstitial expansion. Expanded interstitial compartment compensates sodium accumulation in the extracellular volume due to inappropriate renal sodium retention. Renal sodium retention is brought about by an activation of the molecular structures responsible for the reabsorption of sodium along the cortical collecting duct: amiloride-sensitive epithelial sodium channel at the apical face and sodium pump at the basolateral face of the principal cell. This activation is independent of aldosterone and vasopressin. The asymmetry of expansion between interstitium and plasma compartments is due to impaired Starling forces and increased fluid transfer through the capillary wall. The lack of significant changes in transcapillary oncotic and hydrostatic gradients suggests that increased hydraulic conductivity due to transconformation of endothelial intercellular junctions drives the leakage of fluid into the interstitium and allows to understand the mobility of nephrotic edema. Consistently with the site of renal sodium retention and the activation of the epithelial sodium channel, the association of amiloride and furosemide is efficient to increase urinary sodium excretion, to reverse sodium balance and to remove edema from patients with nephrotic syndrome. PMID- 15351001 TI - [Kangaroo mother care: bibliographical review on the current attitudes, their interests and their limits]. AB - Initiated in 1978 by a Colombian team, then largely adapted in industrialized countries as well as in poor developed countries, the kangaroo mother care (KMC) are known to ensure for low birth weight newborn, a thermoregulation, a good physiological stability and a better relational comfort with their parents. The goal of this work is to make a bibliographical review on current concepts, interests and limits of this method. We re-examined impact of the KMC on the basal metabolism, thermoregulation, growth and evolution of these children. They are helpful in the developing countries but medical safety should not be forgotten. In these countries where there's high frequentation of the services, they are able to regulate body temperature and metabolic adaptation of the newborn. In developed countries, KMC contribute to decrease anxiety of parents and improve the relations with their child. However, it is difficult to recommend their use in current practice. Rigorous randomised studies are necessary to argue their establishment in full safety, to know the neuropsychological development and the real somatic growth on the long term of the children and to known their true economic cost. PMID- 15351002 TI - [A tumour of the right forefinger]. PMID- 15351003 TI - [Sleep problems and bedtime routines]. PMID- 15351005 TI - [Prevention of fetal alcohol syndrome]. PMID- 15351006 TI - [Secondary case of meningitidis after prophylaxis in household contact of meningococcal disease]. PMID- 15351007 TI - [Acceptability and tolerance of prednisolone metasulfobenzoate in orally dispersing tablets in 2 to 12-years-old children]. PMID- 15351008 TI - [A revised nomenclature for allergy]. AB - We expose you the revised nomenclature for allergy following the proposal of the European Academy of Allergy and Clinical Immunology (EAACI). We will review here, as faithfully as possible, the distinct definitions and the right terms to use in allergy and in the other medical branches. PMID- 15351009 TI - [Neonatal death of a newborn twin]. AB - In France, in 2001, 33,000 children were born from multiple pregnancies (4.2% of births). Lately, with the stimulated fertilization improvement, this number has strongly increased. These pregnancies are pretty often difficult and the hospitalizations of the twins (or other multiples) in a neonatal intensive care unit are more frequent than in a single pregnancy (48 vs. 5.3%). Newborn twin death leads the parents to face a tough mourning because of the surviving twin presence. The psychiatrist's function is crucial with the parents, the deceased child, the surviving child and the pediatric staff. Therefore, as much as possible, the psychiatrist (or the psychologist) has to gather and work through the confused feelings of the parents. Several splittings occur in this situation. Some of these splittings concern the medical staff, some others affect the children, the dead one and the living one, either rejected and disinvested or idealized and overprotected. The child psychiatrist is a preferred recipient for the different projections of the parents.The hospital staff as a whole has to understand their full significance and their important psychic sparing for the couple. As the deceased child, the psychiatrist may look bad and unsatisfying, for instance: "he doesn't answer as we would like", "he doesn't come up to the couple's expectations", "he doesn't talk enough", "he's leaving too soon", etc.In addition, the psychiatrist may be attacked as the representative of the pediatric staff, in order to preserve in a better way the others medical protagonists, still essential for the survival of the alive child. These parents are in mourning and the psychiatrist must help them in this process in order to improve the specific investment of the surviving child, the attachment and the communication with him. PMID- 15351010 TI - [Longitudinal study of anthropometric measurements in Parisian children aged ten months to 18 years]. AB - AIM: To describe growth parameters in children followed longitudinally from 10 months of age to 18 years and to compare these data with reference values obtain in children born 30 years earlier. SUBJECTS AND METHOD: A follow study started in 1985 in Paris Health Centres at the ages of I 0 months, 2 and 4 years and subsequently at home every 2 years. Anthropometric data were compared with reference values derived from the French sample of the International Longitudinal Growth study which started in 1953-59. RESULTS: As compared to the reference values, children were taller. At the age of 18 years, in boys, height increase was 5.6 cm. This difference appeared as early as at the age of 12 years. Height increase in girls was 1.6 cm, but this difference was greater at 12. Thereafter, height gain was smaller than 30 years before. The Skelique index was greater due to longer leg length. Fat mass was higher and displayed a more android pattern. A higher prevalence of overweight appeared from the age of 8 years. CONCLUSION: Nowadays, growth processes seem unfavourable as compared to 30 years earlier. Fast growth, long leg length, android body fat distribution and overweight are associated with risk factors of various pathologies(cardiovascular diseases and cancer). Factors promoting these changes over time deserve to be explored more fully. PMID- 15351011 TI - [Evaluation of varicella complications through a retrospective hospital survey in a paediatric center over 16 years in France]. AB - OBJECTIVE: Evaluation of the varicella severity through a prevalence study of hospital admissions justified by a complication directly related to the onset of an acute episode of varicella. METHODS: Retrospective study in one paediatric center in France with a follow-up of a paediatric cohort from April 1987 to December 2002. This general paediatric hospital recruits children from a 400,000 inhabitants area. Inclusion criterion: diagnosis main or associated of varicella. EXCLUSION CRITERIA: congenital or acquired immunodepression, including long-term oral high dosage steroid therapy. RESULTS: Three hundred and forty-three (343) complications of varicella were reported in 309 children hospitalised for a symptom in relationship with varicella. Most of children (75%) were <2 years of age. The annual number of hospitalisations varied with a maximum of 44. An increase of the number of hospitalisations was reported since 1997 but particularly since 2000. Main complications were gastro-intestinal (75), neurological (68), bronchopulmonary (52), and skin and soft tissue infections (52). Superinfections of soft tissues such as Streptococcus pyogenes cellulitis and life-threatening complications, which occurred in children treated by a short course of corticosteroids for an acute episode such as an asthma crisis, were mostly noticed since 1995. Two deaths occurred. CONCLUSION: These data collected over 16 years as part of a retrospective survey of a paediatric cohort show that varicella, often considered as a mild disease, can be responsible for severe complications in young immunocompetent children. The digestive complications (30%) are the main complication in our study with existence of gingivitis stomatitis but also lower digestive manifestations (erosive gastritis aspect through endoscopy). These data have all the more to be taken into account because a vaccine, developed for healthy children, is now available in France. A national survey of hospitalised varicella has been set up since March 2003. PMID- 15351012 TI - [Radiological case study]. PMID- 15351013 TI - Afebrile seizures associated with minor infections: comparison with febrile seizures and unprovoked seizures. AB - This study aimed to demonstrate that afebrile seizures provoked by minor infections constitute a distinct epilepsy syndrome different from febrile seizures and unprovoked afebrile seizures. Of the children who were admitted to hospitals for their first seizure, 1170 had febrile seizures, 286 had provoked seizures, and 125 had unprovoked afebrile seizures. Children with provoked seizures were afebrile at the time of seizure but manifested definite symptoms or signs of minor infection, for example, cough, coryza, vomiting or diarrhea, normal metabolic and cerebrospinal fluid investigations, and no obvious cause for their seizures. The average follow-up was 6.1 years. The Kaplan-Meier estimate of risk at 5 years for subsequent unprovoked afebrile seizures after a first febrile seizure, provoked seizure, or unprovoked afebrile seizure was 1.6%, 5.7%, and 65.7% respectively. All differences were statistically significant (P < 0.0014). In conclusion, afebrile seizures provoked by minor illnesses constitute a distinct type of situation-related seizures, which have not been previously described. Children with provoked seizures have a much lower risk of subsequent unprovoked afebrile seizures than patients with the first afebrile seizure. Careful inquiry for symptoms of minor infections when children present with their first afebrile seizure will help determine the risk for subsequent seizures and the need for antiepileptic drugs. PMID- 15351014 TI - Seizures complicating infantile and childhood bacterial meningitis. AB - In this study, 116 patients, at least 1 month of age but younger than 5 years, were identified with culture-proven bacterial meningitis. A comparison was made between the clinical data of the patients with and without seizures during hospitalization. Seizures during acute bacterial meningitis accounted for 47% (55/116) of the episodes. Time interval between the onset of bacterial meningitis and that of seizures was 1 to 20 days (mean, 4 days). Twelve of these 55 patients had one or more afebrile seizures after completing the treatment. At follow-up of at least 1 year after completing treatment, 26 patients had good outcomes, whereas the other 29 patients had poor outcomes. A strong correlation between the findings of abnormalities through neuroimaging and the occurrence of seizures during hospitalization was observed. The long-term outcomes of patients with infantile and childhood bacterial meningitis, who had seizures during the acute phase of bacterial meningitis, were worse than the outcomes of those who did not have such seizures. No child developed late seizures unless there were acute seizures. Factors associated with seizures during acute bacterial meningitis include disturbed consciousness on admission, abnormal neuroimaging findings, and low glucose and high concentration of total proteins in cerebrospinal fluid. PMID- 15351015 TI - Magnetic resonance imaging in 122 children with spastic cerebral palsy. AB - The interrelationship between magnetic resonance imaging findings, types of cerebral palsy, and gestation was studied. We analyzed the magnetic resonance imaging of brain in 122 children with spastic cerebral palsy. Forty-three patients had spastic hemiplegia, 61 had spastic diplegia, and 18 had spastic tetraplegia. Magnetic resonance imaging abnormalities were observed in 75% of patients. Periventricular leukomalacia accounted for 66% of abnormalities observed in patients with spastic diplegia; other types of brain lesions were uncommon. In patients with spastic tetraplegia, two types of magnetic resonance imaging abnormalities predominated: congenital brain anomalies and term-type brain injuries, 42% and 33% respectively. Types of magnetic resonance imaging abnormalities were more heterogeneous in patients with spastic hemiplegia. Preterm brain injuries (periventricular leukomalacia and posthemorrhagic porencephaly) were observed often in patients born at preterm but were also observed in patients born at term. Term-type brain injuries (term-type border zone infarct, basal ganglia-thalamic lesion, subcortical leukomalacia, and multicystic encephalomalacia) were observed only in patients born at or near term. We conclude that magnetic resonance imaging findings for patients with spastic cerebral palsy were closely related to types of cerebral palsy and gestation at birth. Magnetic resonance imaging in patients with perinatal brain injury may reflect pathologic changes and is useful in understanding and evaluating cerebral palsy. PMID- 15351016 TI - Gastric emptying in children with cerebral palsy and gastroesophageal reflux. AB - Gastric emptying time is considered a factor in the increased frequency of gastroesophageal reflux in children with cerebral palsy. It is unknown if emptying time influences the severity of reflux. In this study, 76 cerebral palsy patients with reflux indicative symptoms were investigated by 24-hour pH monitoring. Reflux complications were also studied. Emptying time in children with reflux was investigated using gastric scintigraphy. Twenty-eight children with resistant asthma scanned for pulmonary aspiration were studied as control subjects for emptying time. Reflux was diagnosed in 51.3%; it was severe in 53.8%, moderate in 38.5%, and mild in 7.7%. Occurrence of reflux did not differ significantly among different forms of cerebral palsy or between males and females. The most frequent complications in reflux-positive patients were iron deficiency (51.3%), anemia (41.0%), malnutrition (33.3%), recurrent upper respiratory tract infections (28.2%), and low body weight (28.2%). Patients without reflux had less frequent complications. Gastric emptying time measured by gastric scintigraphy in 28 patients with reflux manifested no difference in comparison to the control group (P > 0.05). No relationship was found between emptying time and reflux severity (P > 0.05). In conclusion, reflux (moderate or severe) is common in children with cerebral palsy, frequently leading to complications but no delayed emptying time. The patients described in this report had no delayed emptying time. There was also no relationship between emptying time and severity of reflux. PMID- 15351017 TI - Expression of genes related to muscular dystrophy with lissencephaly. AB - There is a group of congenital muscular dystrophies accompanying the brain lesions termed cobblestone lissencephaly. Abnormal glia limitans could be considered the major pathogenesis of cobblestone lissencephaly. In this group, protein-O-linked mannose-beta1,2-N-acetylglucosaminyltransferase and protein-O mannosyltransferase 1 are considered to be responsible for muscle-eye-brain disease and Walker-Warburg syndrome, respectively, by glycosylation of alpha dystroglycan. However, the functions of fukutin, a gene responsible for Fukuyama type congenital muscular dystrophy, are still unclear. In this study, expression of the three aforementioned genes was compared by in situ hybridization in control cases to elucidate the functions of fukutin. Immunohistochemistry of fukutin and alpha-dystroglycan was also performed. In the central nervous system, all three genes were expressed in astrocytes and in immature neurons. A few mature neurons expressed fukutin, but many expressed the other two genes. All genes were expressed in various non-nervous tissues including tissues relating to secretion. Fukutin and alpha-dystroglycan were generally colocalized, but localization was not always the same, especially in the liver. Fukutin may be associated with the glycosylation of alpha-dystroglycan, and expression in astrocytes may indicate a relation to glia limitans. The roles of fukutin in mature neurons may be less critical compared with the other two genes. Additional functions of fukutin, especially in the liver, are suspected. PMID- 15351018 TI - Neuropsychological outcome after acute disseminated encephalomyelitis: impact of age at illness onset. AB - Recovery from acute disseminated encephalomyelitis in childhood appears relatively uneventful, at least when looking at functional recovery parameters such as neurologic outcome. However, neuropsychology literature suggests that relatively transient illnesses affecting the central nervous system are associated with cognitive and social sequelae, particularly when the illness occurs during the preschool years. This study investigated the impact of timing of acute disseminated encephalomyelitis on intellectual, educational, and social functioning in children. Nineteen children (10 with acute disseminated encephalomyelitis before the age of 5 years), who had been admitted to the Royal Children's Hospital, Melbourne Australia, in the past 6 years underwent a brief neuropsychologic assessment. Performance was compared with 19 control subjects, stratified for age and socioeconomic status with the acute disseminated encephalomyelitis group. Children who sustained their illness before 5 years of age were particularly vulnerable to impairments in both cognitive and social domains. In particular, a higher incidence of severe behavioral and emotional problems was reported by parents of children who had experienced acute disseminated encephalomyelitis before 5 years of age. This finding suggests that there may be long-term complications in early childhood. A multidisciplinary approach to management post-illness is warranted in this age group. PMID- 15351019 TI - Ketogenic diet: outpatient initiation, without fluid, or caloric restrictions. AB - Although the ketogenic diet has been used for more than 80 years, the optimal methods of initiating the diet and its maintenance have not been clearly defined. This retrospective study was performed to review our experience with initiation of the ketogenic diet in the outpatient and inpatient settings and maintenance of the diet without fluid or caloric restriction. We analyzed 54 patients who had medically intractable epilepsy of whom 44% manifested some degree of mental retardation, 80% had multiple seizure types, and failed on average 4.8 antiepileptic drugs. Forty-four patients underwent induction of the ketogenic diet on an outpatient basis and 21 as inpatients. Three patients in each group were fasted at the initiation of the diet. No significant differences were observed with regard to seizure control in that 62% and 71% had greater than 50% improvement in the outpatient and inpatient groups, respectively. Both groups manifested improvement in alertness and social interaction. The efficacy of a ketogenic diet in the symptomatic epilepsies was confirmed, and benefit for medically refractory childhood absence epilepsy was documented. We conclude that a prospective, randomized trial is necessary to compare outpatient vs inpatient initiation of the ketogenic diet and the utility of fluid and caloric restriction. PMID- 15351020 TI - Diffusion tensor imaging in frontal lobe epilepsy. AB - We report a 13-year-old female with refractory frontal lobe epilepsy in whom diffusion tensor imaging was useful for exploring subtle cortical malformation. She had frequent simple partial seizures characterized by clonic movement of the right upper extremity. Conventional magnetic resonance imaging was not conclusive. A diffusion tensor imaging revealed an increased apparent diffusion coefficient and reduced anisotropy in the left frontal lobe. Positron emission tomography disclosed hypermetabolism in the adjacent area. We could identify the relatively thick cortex with an indistinct gray-white matter junction in a thin slice T(1)-weighted image. PMID- 15351021 TI - A magnetoencephalographic study of astatic seizure in myoclonic astatic epilepsy. AB - To study the pathophysiologic mechanism of astatic seizures in a patient with myoclonic astatic epilepsy of early childhood, ictal magnetoencephalography was recorded and the neuronal pathway involved was analyzed. The patient was a 12 year-old female who developed myoclonic and astatic seizures including nodding and sudden falling at the age of 4. The current source of spikes during nodding attacks was located in the bilateral frontal area with left predominance, possibly in the premotor cortex. Although we could not claim, on the basis of our findings, that myoclonic astatic epilepsy of early childhood is a type of focal epilepsy, it seems likely that the premotor cortex might be more excitable than other areas. Thus we speculate that the functionally altered premotor reticulospinal tract which normally controls postural adjustment might play an important role in the generation of myoclonic astatic seizures. Furthermore, the underlying mechanism in the brainstem seems to be common, at least in part, for infantile spasms when considering the efficacy of synthetic adrenocorticotropic hormone for nodding seizures. PMID- 15351022 TI - Pyridoxine-dependent seizures and microcephaly. AB - Pyridoxine dependency is a rare autosomal-recessive disorder causing intractable seizures in neonates and infants. This case report describes an infant with pyridoxine-dependent seizures with microcephaly and discusses a probable pathogenetic mechanism of microcephaly in this condition. PMID- 15351023 TI - Subacute sclerosing panencephalitis: a cause of acute vision loss. AB - A 4-year-old male presented with only acute vision loss. His neurologic examination, funduscopic examination, and pupils were normal. Cranial magnetic resonance imaging revealed abnormal hyperintense, bilaterally symmetric lesions (on T(2)-weighted and fluid-attenuated inversion recovery, images) in bilateral optic radiations, pulvinar region in the thalami, crus posterior of internal capsules, periventricular white matter, and unilaterally left anterior pons. Elevated measles antibody titers in the cerebrospinal fluid confirmed the diagnosis of subacute sclerosing panencephalitis. Vision loss improved and cranial magnetic resonance imaging findings regressed,but myoclonic jerks and deterioration began 7 months later. The diagnosis of subacute sclerosing panencephalitis should be considered in cases with acute vision loss resulting from cortical blindness even when classical findings of the central nervous system do not exist. PMID- 15351024 TI - Riboflavin-responsive glutaric aciduria type II with recurrent pancreatitis. AB - A 22-year-old woman had suffered from several episodes of acute pancreatitis since the age of 11. Other than exercise intolerance since early childhood, her psychomotor development was normal. At age 21, she experienced two episodes of generalized muscle weakness including acute respiratory failure and hepatomegaly. Liver biopsy indicated fatty metamorphosis, and muscle biopsy revealed vacuolar myopathy with lipid accumulation. Biochemical investigations demonstrated elevated serum creatine kinase and elevated 2-hydroxylglutaric, pyruvic, ethylmalonic, hippuric, adipic, and seburic acids in urinary organic acid analysis. These findings confirmed the diagnosis of glutaric aciduria type II. Although acute pancreatitis in glutaric aciduria type II has been reported previously, this is the first reported case of recurrent pancreatitis occurring in glutaric aciduria type II. We treated the patient with l-carnitine and riboflavin. As of the latest follow-up 2.5 years later, the patient has had no further episodes of muscle weakness or pancreatitis. We suggested analyzing urine organic acid when lipid storage myopathy is suspected. PMID- 15351025 TI - Acute fulminant subacute sclerosing panencephalitis with absent measles and PCR studies in cerebrospinal fluid. AB - This report describes an atypical case of rapidly progressive subacute sclerosing panencephalitis presenting as transient visual agnosia and myoclonus in a 14-year old male. There were no typical periodic complexes in serial electroencephalographic monitoring; cerebrospinal fluid measles antibody titer was negative. The diagnosis was made by molecular and histologic examination of open brain biopsy tissue. PMID- 15351026 TI - Intrauterine shoulder weakness and obstetric brachial plexus palsy. AB - Obstetrical brachial plexus injury occurs when the forces preventing the stretch of the brachial plexus are overcome by the forces stretching it. This report describes an 8-day-old male delivered by uncomplicated cesarean section with right obstetrical brachial plexus palsy and congenital arm atrophy. The patient had a history of decreased right arm movement detected by fetal ultrasound at 18 to 20 weeks of gestation. The purpose of this article is to report that stretching of brachial plexus at birth sufficient to produce a plexus injury may occur in a patient with a vulnerable plexus even in the absence of traction during delivery. PMID- 15351027 TI - Glutaric aciduria type 1: proton magnetic resonance spectroscopy findings. AB - Glutaric aciduria type 1 is an inborn error of lysine, hydroxylysine, and tryptophan metabolism caused by deficiency of glutaryl-coenzyme A dehydrogenase. The disease often appears in infancy with an encephalopathic episode that results in acute basal ganglia and white matter degeneration. The neuroimaging findings in glutaric aciduria type 1 have been well defined. However, the changes in magnetic resonance spectroscopy, a noninvasive tool for identifying the biochemical state of the brain, are scarce in glutaric aciduria type 1. This report presents the magnetic resonance spectroscopy findings in a 19-month-old male with glutaric aciduria type 1. Magnetic resonance spectroscopy of right frontal white matter and right lentiform nuclei revealed decreased N acetylaspartate/creatine ratio, slightly increased choline/creatine ratio, and increased myoinositol/creatine ratio, compared with the age-matched control patients. We thought that these changes were in accordance with neuroaxonal damage, demyelination, and astrocytosis in these areas. In conclusion, proton magnetic resonance spectroscopy provides a tool for assessing metabolic disturbances and the extent of brain damage noninvasively in glutaric aciduria type 1. PMID- 15351028 TI - Meropenem decreases serum level of valproic acid. AB - Concomitant administration of meropenem has been reported to decrease serum level of valproic acid both in humans and in animals. This report describes three children who required simultaneous administration of valproic acid and meropenem. Meropenem rapidly decreased serum valproic acid concentration to subtherapeutic levels in all three children, and serum valproic acid levels were returned to therapeutic levels in a short time after discontinuing simultaneous meropenem therapy. Valproic acid was not changed to another antiepileptic agent, because no seizure activity was observed. To our knowledge, this is the first case report on the simultaneous administration of meropenem and valproic acid in childhood. In conclusion, it is clear that concomitant meropenem administration decreases serum valproic acid concentration, and we believe that there may be no need to change the antiepileptic drug during this period, provided that the patient has no seizure. More detailed studies are required. PMID- 15351029 TI - Photoparoxysmal responses in children: their characteristics and clinical correlates. PMID- 15351031 TI - Brain calcification in Mobius syndrome. PMID- 15351033 TI - Internalization of non-toxigenic Corynebacterium diphtheriae by cultured human respiratory epithelial cells. AB - Although infection by Corynebacterium diphtheriae is a model of extracellular mucosal pathogenesis, and diphtheria is one of the most worried diseases, this microorganism can be associated also with invasive infections such as endocarditis, septic arthritis, and osteomyelitis. Invasive infections are usually caused by non-toxigenic C. diphtheriae strains. Over the last years severe pharyngitis/tonsillitis associated with the isolation of non-toxigenic C. diphtheriae have been described. Penicillin treatment failure of these infections could only partially be explained by penicillin tolerance of the causing strain. Thus, we examined the in vitro ability of non-toxigenic C. diphtheriae throat clinical isolates to adhere to, and enter human respiratory epithelial cells. Trasmission and scanning electron microscopy demonstrated intracellular C. diphtheriae in laryngeal (HEp-2 cells) and pharyngeal (Detroit D562 cells) tissue culture. Live intracellular bacteria were detectable up to 48 h post-infection. Using a variety of compound that act on eukariotic cell structures, the internalization of C. diphtheriae seems to occur via a zipper-like mechanism. It is likely that internalization of C. diphtheriae can be involved in throat colonization contributing to bacterial eradication failure and asymptomatic carriage. PMID- 15351034 TI - Tumor necrosis factor-alpha is required for gastritis induced by Helicobacter felis infection in mice. AB - Helicobacter pylori colonizes the gastric mucosa of human and causes chronic gastritis. The previous studies have demonstrated that gamma interferon (IFN gamma) but not tumor necrosis factor-alpha (TNF-alpha) plays a critical role in pathogenesis of gastritis induced by H. pylori infection. In this study we investigated the induction of gastritis induced by H. felis infection in TNF alpha-deficient mice, comparing with IFN-gamma-deficient mice. The scores of gastritis and epithelial changes of TNF-alpha-deficient mice and IFN-gamma deficient mice were significantly lower than that of C57BL/6 mice. Moreover, the degrees of gastritis and epithelial changes of TNF-alpha-deficient mice were rather low compared with that of IFN-gamma-deficient mice. In spleen cell cultures stimulated with heat-killed H. felis, IFN-gamma production by TNF-alpha deficient mice and TNF-alpha production by IFN-gamma-deficient mice were significantly reduced compared with those in C57BL/6 mice. These results suggested that TNF-alpha is involved in pathogenesis of gastritis induced by H. felis infection as IFN-gamma and that an interaction between TNF-alpha and IFN gamma might be required in pathogenesis of gastritis induced by Helicobacter infection. PMID- 15351035 TI - Role of bovine herpesvirus 4 in bacterial bovine mastitis. AB - In order to study the role played by bovine herpesvirus 4 (BoHV-4) in bovine mastitis, PCR experiments were performed on a Hungarian dairy herd of 2000 cows. Milk cells were tested with a nested PCR adjusted to detect the virus in the milk. Thirty to forty-one percentage of the udders of 101 cows with bacterial mastitis (Escherichia coli, Streptococcus uberis or Staphylococcus aureus) gave positive results, whereas less than 6% of the milk samples were positive for BoHV 4 from 118 animals with healthy udders. The mastitis status of these 118 healthy cows was followed throughout the milking period; 4.2% had clinical, and 25.42% had subclinical mastitis. By the end of the milking period, more than 90% of the cows shed the virus in their milk, regardless of the bacterial status of the udder. No correlation was found between the virus shed, the somatic cell count, and the bacterial status of the udder. Viral DNA was detected in the wall of the milk duct. These results demonstrate that BoHV-4 neither causes mastitis directly nor plays a role in the initiation of the process, but later, when bacterial infection of the udder occurs, the reactivated virus replicates in the immune cells of the udder and/or in the epithelial cells of the milk ducts and may be responsible for more severe, prolonged mastitis. As mastitis is a crucial problem of milk production, this virus may be considered a possible predisposing factor and also an agent of secondary udder infections in prolonged mastitis cases. PMID- 15351036 TI - Immunofluorescence and flow cytometry analysis of fibronectin and laminin binding to Sporothrix schenckii yeast cells and conidia. AB - The adherence of Sporothrix schenckii yeast cells to several extracellular matrix (ECM) components has already been demonstrated, but the mechanisms of these interactions remained to be defined. In indirect immunofluorescence assays with polyclonal antibodies directed towards the ECM proteins, both hyphae and yeast cells of S. schenckii exhibited the ability to bind laminin and fibronectin. Flow cytometry confirmed the binding of these proteins, and revealed a significant greater binding capability for the yeast cells than for the conidia. Fibronectin and laminin binding was dose-dependent and specific. In addition, competition experiments with synthetic peptides mimicking the adhesive sequences of these proteins, or with cell wall fractions and carbohydrates constitutive of their sugar chains, were performed in order to specify the peptide or carbohydrate motifs involved in the recognition process. A 50% reduction was noticed in fibronectin binding in the presence of the synthetic peptide RGD, and a 38% reduction in laminin binding with the peptide YIGSR. Some carbohydrate-containing fractions of the yeast cell wall also inhibited the binding of fibronectin, but had no significant effect on laminin binding. Together, these results suggest the presence at the yeast surface of distinct receptors for laminin and fibronectin. PMID- 15351037 TI - Tobacco smoke induces persistent infection of Chlamydophila pneumoniae in HEp-2 cells. AB - We examined tobacco smoke exposure and its effect on the life cycle of Chlamydophila pneumoniae (C. pneumoniae) in HEp-2, a human respiratory epithelial cell line. Using noncytotoxic concentrations of smoke medium, chlamydiae were grown in tissue culture and infectious particles were quantitated indirectly by immunocytometry of infected indicator cells. Chlamydial genome copy number was assessed with real-time polymerase chain reaction, and ultrastructure was examined by transmission electron microscopy. There was a significant reduction (56-64%; p<0.05) in the number of infectious elementary bodies following smoke exposure compared to untreated cultures. Under the same conditions, at late time points, smoke-exposed cultures showed significantly fewer chlamydial DNA copies (p<0.04). Moreover, smoke exposure induced large aberrant bodies that predominated within the inclusion. Following in vitro smoke exposure, alterations in the developmental cycle of C. pneumoniae included: inhibition of productive infection, reduced bacterial cell division, and formation of aberrant bodies. Thus, using this novel system, we were able to induce chlamydial persistence. Tobacco smoke exposure may represent a risk for establishment of a chronic reservoir of C. pneumoniae infection within respiratory epithelium. PMID- 15351038 TI - The NheA component of the non-hemolytic enterotoxin of Bacillus cereus is produced by Bacillus anthracis but is not required for virulence. AB - A non-hemolytic enterotoxin (NHE) is one of the two enterotoxins thought to cause diarrhea produced by Bacillus cereus. We identified genes in Bacillus anthracis homologous to the B. cereus nheAB genes encoding proteins of the NHE complex. The NheA component was detected immunologically in culture supernatants from B. anthracis but not from a NheA(-) mutant, suggesting that B. anthracis produces and secretes the NheA subunit of NHE. A NheA deletion mutant was not attenuated in the guinea pig suggesting that NheA is not absolutely required for virulence. PMID- 15351039 TI - Production of prostaglandin E2 in monocytes stimulated in vitro by Chlamydia trachomatis, Chlamydophila pneumoniae, and Mycoplasma fermentans. AB - Chlamydia trachomatis (CT) as well as Chlamydophila pneumoniae (CP) cause chronic inflammatory diseases in humans. Persistently infected monocytes are involved in the pathogenesis by inducing mediators of inflammation. An in vitro system of chlamydial persistence in human peripheral blood monocytes (HPBM) was used to investigate prostaglandin E(2) (PGE(2)) production and the expression of the key enzyme for prostaglandin production, cyclooxygenase-2 (COX-2). PGE(2) production was determined by PGE(2)-ELISA of HPBM-culture supernatants. Cox-2 mRNA expression was measured by real-time RT-PCR of total RNA isolated from HPBM. Both, CT and CP, stimulated PGE(2) production of HPBM in vitro. Equivalent numbers of CT per host cell induced a higher PGE(2)-response compared to CP. The amount of synthesized PGE(2) depended on the chlamydial multiplicity of infection (MOI). Even at an MOI of 10 the amount of CT- and CP-induced prostaglandin, respectively, was lower than the amount of prostaglandin induced by E. coli lipopolysaccharide (LPS) at a concentration of 10microg/ml. In contrast to stimulation with LPS, Chlamydia-induced PGE(2) production as well as cox-2 mRNA decreased after day 1 post infection (p.i.). These data indicate that Chlamydia stimulate PGE(2) production in human monocytes. Since Chlamydia are often contaminated by mycoplasma, the influence of mycoplasma on the prostaglandin production was investigated additionally. Mycoplasma fermentans (MF) also stimulated PGE(2) production. The co-infection of mycoplasma and Chlamydia resulted in an additive effect in the production of PGE(2). Thus it is important to use host cells and Chlamydia free of mycoplasma contamination for the analysis of Chlamydia-induced prostaglandin production. PMID- 15351040 TI - Quantitation of H. pylori cytotoxin mRNA by real-time RT-PCR shows a wide expression range that does not correlate with promoter sequences. AB - Up to 28-fold differences in vacA expression in Helicobacter pylori strains grown in vitro were demonstrated by real time quantitative RT-PCR. These large differences in expression were unrelated to putative -35 and -10 motifs or to other untranslated sequences upstream of the ATG start site. The lack of correlation between promoter sequences and the vacA expression levels suggest the potential existence of a bacterial strain-specific factor, as earlier proposed by others on the basis of reporter gene fusions. PMID- 15351041 TI - A novel double coating for microemulsion electrokinetic chromatography with laser induced fluorescence detection: as tested with amino acid derivatives. AB - A novel double coating (DC) was developed for fast and reproducible microemulsion electrokinetic chromatography (MEEKC), as tested with separation and determination of amino acids using laser-induced fluorescence (LIF) detection after derivatization with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazol. The simple coating is a combination of a removable covalent layer and a dynamic SDS coating. Hexamethyldisilazane was utilized for the covalent layer that can be regenerated on-line. Compared with previous no-coating method, the analysis time was shortened; and the reproducibility of migration times was improved. PMID- 15351042 TI - Electrophoretic analysis of di- and oligosaccharides derived from glycosaminoglycans on microchip format. AB - Microchip electrophoresis is a powerful tool for fast analysis of nucleic acids and has expanded its applicability to the analysis of various biological materials including proteins and carbohydrates. Glycosaminoglycans have intrinsic negative charges, and are good targets for electrophoretic analysis. In the present paper, we developed a method to analyze oligosaccharides and unsaturated disaccharides derived from some glycosaminoglycans after digestion with specific enzymes followed by derivatization with 2-aminoacrydone (AMAC) by reductive amination. The method described here allowed rapid analysis of oligosaccharides derived from glycosaminoglycans within 150 s with high sensitivity. We show an application of the present technique to the glycosaminoglycan analysis in cultured HeLa cells. PMID- 15351043 TI - A rapid and simple determination of A77 1726 in human serum by high-performance liquid chromatography and its application for optimization of leflunomide therapy. AB - Leflunomide is a disease-modifying antirheumatic drug, which is bioactivated by formation of A77 1726. In this study a rapid and simple quantitative assay using a reversed phase HPLC-UV method is validated for detection of A77 1726 in human serum. The HPLC-UV method uses a mobile phase consisting of methanol and a KH2PO4 buffer (45 mM, pH = 3) (50:50,v/v), at a flow rate of 1 mL/min. A77 1726 is detected by UV-absorption at 295 nm with a retention time of 8.9 min. Demoxepam is used as internal standard. Validation showed lower and upper limits of quantitation of 0.5 and 100 mg/L, respectively. The assay was linear over the concentration range of 0.5-100 mg/L (r2 > 0.999). Intra- and inter-day precision showed coefficients of variation within 15% over the complete concentration range; accuracy was within 8%. Commonly prescribed drugs to treat rheumatoid arthritis like disease-modifying antirheumatic drugs, analgesics and corticosteroids, and their main metabolites, are separated from A77 1726 with a resolution >2. Serum levels of A77 1726 in 37 patients on leflunomide therapy were determined using this HPLC-UV method. Measured serum A77 1726 serum concentrations in patient samples showed large variability with a range of 3-176 mg/L. PMID- 15351044 TI - HPTLC method for guggulsterone. II. Stress degradation studies on guggulsterone. AB - Stress degradation studies were carried out on guggulsterone (the hypolipidemic agent in the gum-resin exudates of Commiphora mukul) following the conditions prescribed in the parent drug stability testing guideline (Q1AR) issued by International Conference on Harmonization (ICH). The present study describes degradation of guggulsterone under different ICH prescribed stress conditions (acid and base hydrolysis, oxidation, dry and wet heat degradation and photodegradation) and establishment of a stability indicating HPTLC assay. The method employed TLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of toluene-acetone (9:1, v/v). Densitometric analysis of guggulsterone was carried out in the absorbance mode at 250 nm. This system was found to give compact spots for E- and Z-guggulsterone, (Rf value of 0.38 +/- 0.02 and 0.46 +/- 0.02, respectively) following double development of chromatoplates with the same mobile phase. The drug undergoes degradation under acidic and basic conditions, oxidation, dry and wet heat treatment and photodegradation. All the peaks of degraded products were resolved from the standard guggulsterone with significantly different Rf values. As the method could effectively separate the drug from its degradation products, it can be employed as a stability indicating one. PMID- 15351045 TI - HPTLC method for guggulsterone. I. Quantitative determination of E- and Z guggulsterone in herbal extract and pharmaceutical dosage form. AB - A sensitive, selective, precise and robust high-performance thin-layer chromatographic method of analysis of E and Z stereoisomers of guggulsterone (the hypolipidemic agent in the gum-resin exudates of Commiphora mukul) both as a bulk drug and in formulations was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of toluene-acetone (9:1, v/v). Densitometric analysis of guggulsterone was carried out in the absorbance mode at 250 nm. This system was found to give compact spots for E- and Z-guggulsterone (Rf value of 0.38 +/- 0.02 and 0.46 +/- 0.02, respectively) following double development of chromatoplates with the same mobile phase. The linear regression analysis data for the calibration plots for E- and Z-guggulsterone showed good linear relationship with r2 = 0.9977 +/- 0.054 and 0.9975 +/- 0.068, respectively, in the concentration range of 100-6000 ng/spot. The mean value of slope and intercept were 0.11 +/- 0.006 and 0.11 +/- 0.005, 14.26 +/- 0.56 and 10.92 +/- 0.76, respectively, for E- and Z-guggulsterone. The method was validated for precision, robustness and recovery. The limit of detection and quantitation were 12, 10 and 24, 20 ng/spot, respectively, for E- and Z-guggulsterone. Statistical analysis proves that the method is repeatable and selective for the estimation of the said drug. Since the proposed mobile phase effectively resolves the E- and Z-isomers of guggulsterone, this HPTLC method can be applied for identification and quantitation of these isomers in herbal extracts and pharmaceutical dosage form. PMID- 15351046 TI - Improved detection of Coptidis alkaloids by field-amplified sample stacking in capillary electrophoresis. AB - To improve the on-line detection of Coptidis alkaloids in capillary electrophoresis the field-amplified sample stacking was studied for them. In this work the peak height enhancements of stacking with hydrodynamic and electrokinetic injections were compared with respect to the conventional sample injection. It was found that the stacking efficiency of electrokinetic injection was more than ten times greater than that of hydrodynamic injection. No peak height enhancement was observed with the pre-injection of a short water plug before sample injection with electrokinetic injection. The concentration limits of detection of berberine, coptisine and palmatine obtained with electrokinetic injection were about 5 ng/ml (ppb), which was approximately 240 times lower than those from conventional sample injections. Baseline separation was also achieved for the main alkaloids. After validation the developed method was applied to determine the quantity of berberine, coptisine and palmatine in a Coptidis Rhizoma sample. The method is simple, rapid and should be able to be used in identification and quantitative evaluation of the crude drugs. PMID- 15351047 TI - Chloride-selective membrane electrodes and optodes based on an indium(III) porphyrin for the determination of chloride in a sequential injection analysis system. AB - Two quasi-independent methods for potentiometric and optical determination of chloride were simultaneously implemented in a flow system, providing real-time assessment of the quality of results. A potentiometric and an optical polymeric membrane doped with the same indium(III) octaethyl-porphyrin were used as sensor ionophore. The working mechanism and the analytical characteristics of these porphyrin-based sensors with respect to dynamic range, selectivity, repeatability and lifetime are discussed. These sensors, utilised as detectors in a flow system, were applied for the analysis of chloride in pharmaceutical solutions. The quality of the results obtained was evaluated by comparison with those provided by the reference method and no significant statistical differences at the 95% confidence level were observed. The simultaneous attainment of two measurements permitted the standardisation of results in real time and the detection of failures in the procedure. PMID- 15351048 TI - Development and validation of a fully automated method for the chromatographic determination of content uniformity of drug tablets. AB - A fully automated method for the content uniformity analysis of LAS 34475 25mg tablets has been developed by using an automated procedure. This automated method has been validated within the requirements of ICH guidelines Q2A-Q2B. Standard and sample solutions are processed by an automated benchtop system. The operations automated include the phases of disintegration of the dosage form, filtration of the resultant homogenate and injection of the clear sample into the chromatographic system. Although a manual method validated according to ICH guidelines already existed for this compound, the benefits of applying appropriate automation should provide continuous operation, increased precision, an affordable electronic audit trail and significantly reduced time consumption as well as reducing the exposure of the analyst to the drug substance. The objective of this work was to adapt the manual method to an automated workstation. Considerable effort went into developing and validating an automated method. The results obtained in the validation of this automated method were equivalent to the manual method in terms of system precision, linearity, accuracy, robustness and sensitivity (limits of detection, LOD and limits of quantification, LOQ), and carry-over. PMID- 15351049 TI - Method development for betamethasone and dexamethasone by micellar liquid chromatography using cetyl trimethyl ammonium bromide and validation in tablets. Application to cocktails. AB - An isocratic liquid chromatographic method for the determination of betamethasone (BM) and dexamethasone (DM) using methylprednisolone (MPL) as internal standard and micellar mobile phases consisting of cetyl trimethyl ammonium bromide (CTAB) and organic modifiers such as propanol, butanol and pentanol has been developed. The effect of organic modifiers, surfactant concentration, temperature and flow rate on the separation has been studied. Method validation for dexametasone or bethametasone in tablets was carried out using a mobile phase 0.24% pentanol and 32.5 mM CTAB, a flow-rate of 0.5 ml min(-1), an Hypersil C18 column (60 degrees C), and UV detection at 243 nm. The recoveries for BM and DM found in the accuracy test were 99 +/- 3 and 101 +/- 2, respectively. Repeatability and intermediate precision expressed as R.S.D. were lower than 5% for both compounds. The proposed method was applied to cocktails containing both compounds. PMID- 15351050 TI - Development and validation of a novel LC/ELSD method for the quantitation of gentamicin sulfate components in pharmaceuticals. AB - The equivalent response of evaporative light scattering detector (ELSD) for compounds of similar structure is exploited to develop an LC/ELSD method for the simultaneous quantitation of the four main components of gentamicin sulfate, using as external standard the one main component kanamycin. A C18 column was used along with a mobile phase consisting of H2O (containing 35.4 microg/ml of trichloroacetic acid and 0.89 microl/ml of trifluoroacetic acid)-methanol acetonitrile (990:5:5, v/v/v), in an isocratic mode at 1.1 ml/min. Parameters of ELSD were 50 degrees C for evaporation temperature and 3.0 bar for pressure of carrier gas (N2). A logarithmic calibration curve was obtained for sulfate (tR = 1.9 min) from 4.2 to 150 microg/ml (r > 0.994) with a precision of 0.18%R.S.D. Kanamycin and the four gentamicin components (C(1a), C2, C(2a) and C1) were eluted at 3.2, 4.6, 5.9, 7.1 and 8.7 min, respectively, with good resolution (Rs > 1.5). Logarithmic calibration curve was obtained for each component (r > 0.99) with statistically equal slopes varying from 2.457 to 2.558. The mass range of total gentamycin was 35-240 microg/ml. The proposed method was applied for the determination of gentamicin components and sulfate in raw materials and pharmaceutical formulations (injection, drops and cream) without any pretreatment except cream, for which liquid-liquid extraction was required. Recovery from standard addition experiments in commercial formulations was 99-100% regarding total gentamicin and 89-108% regarding individual components, with a precision (%RSD, n = 4) 0.7-5.8%. PMID- 15351051 TI - Identification of 5-aminosalicylic acid, ciprofloxacin and azithromycin by abrasive stripping voltammetry. AB - Solid microparticles of 5-aminosalicylic acid, ciprofloxacin, and azithromycin were mechanically immobilized on the surface of the paraffin impregnated graphite electrode and investigated by square-wave and cyclic voltammetry in order to develop a method for their qualitative determination. 5-Aminosalicylic acid is oxidized at 0.540 V in the quasireversible electrode reaction, which is followed by the chemical transformation of the product, while ciprofloxacin and azithromycin are oxidized at 1.2 V and 0.94 V, respectively, in totally irreversible electrode reactions. The detection of these drugs in commercial dosage formulations is reported. PMID- 15351052 TI - Comparison of fluorimetric, voltammetric and biosensor methods for the determination of total antioxidant capacity of drug products containing acetylsalicylic acid. AB - Antioxidant capacity of several drug specialities containing as mean component acetylsalicylic acid were experimentally evaluated using an enzymatic electrode, recently developed by the present authors, based on superoxide dismutase (SOD) enzyme. The precision of this method of analysis was found to be good (for drug samples RSD < or = 5%). The results were also compared with those ones by a traditional spectrofluorimetric method and by two other methods, respectively, based on cyclic and pulsed voltammetry, recently trialled by the present authors. PMID- 15351053 TI - Study of urinary 8-hydroxydeoxyguanosine as a biomarker of oxidative DNA damage in diabetic nephropathy patients. AB - Increased oxidative stress induced by hyperglycemia may contribute to the pathogenesis of diabetic complications. Urinary 8-hydroxydeoxyguanosine (8-OHdG) has been reported to serve as a sensitive biomarker of oxidative DNA damage and also of oxidative stress. This article studied oxidative DNA damage in patients with diabetic nephropathy and in healthy control subjects by urinary 8-OHdG evaluations. Contents of 8-OHdG in urine were analyzed by capillary electrophoresis with end-column amperometric detection (CE-AD) after a single step solid-phase extraction (SPE). Levels of urinary 8-OHdG in diabetic nephropathy patients with macroalbuminuria was significant higher than in control subjects (5.72 +/- 6.89 micromol/mol creatinine versus 2.33 +/- 2.83 micromol/mol creatinine, P = 0.018). A significant difference of 24 h urinary 8-OHdG excretions exists between the patients with macroalbuminuria and the patients with normoalbuminuria (19.2 +/- 16.8 microg/24 h versus 8.1 +/- 1.7 microg/24 h, P = 0.015). There was a positive correlation between urinary excretion of 8-OHdG and glycosylated hemoglobin (HbA1c) (r = 0.287, P = 0.022). A weak correlation exists between the levels of 8-OHdG and triglyceride (r = 0.230, P = 0.074). However, the urinary 8-OHdG contents are not correlated with blood pressure and total cholesterol. The increased excretion of urinary 8-OHdG is seen as indicating an increased systemic level of oxidative DNA damage in diabetic nephropathy patients. PMID- 15351054 TI - The development of a higher throughput reactive intermediate screening assay incorporating micro-bore liquid chromatography-micro-electrospray ionization tandem mass spectrometry and glutathione ethyl ester as an in vitro conjugating agent. AB - An in vitro reactive intermediate screening assay, incorporating the use of the close analog of glutathione, glutathione ethyl ester (GSH-EE) as a conjugating agent, was developed to identify compounds that form reactive intermediates in an in vitro metabolite generating system. The biological assay consisted of substrate [s] = 10 microM, human liver microsomes, an NADPH generating system and glutathione ethyl ester. Conjugates were extracted from the biological matrix using a combination of protein precipitation and a semi-automated 96-well plate solid phase extraction (SPE) procedure. A micro-bore liquid chromatography-micro electrospray ionization-tandem mass spectrometry (microLC-microESI-MS/MS) method detected glutathione ethyl ester conjugates using selected reaction monitoring (SRM) to simultaneously monitor for multiple MH+ to [MH - 129]+ transitions, where the 129 mass unit (Da) represents the neutral loss of the pyroglutamate moiety from GSH-EE. The multiple MH+ to [MH - 129]+ transitions (SRM mass table) were generated for potential reactive intermediates of each compound. Glutathione (GSH) and GSH-EE conjugate standards were used to evaluate MS detection sensitivity. Based on direct comparison of standard curve data, an approximate 10 fold increase in sensitivity was observed for conjugates containing GSH-EE moiety versus GSH. In vitro experiments were conducted using literature substrates acetaminophen, rosiglitazone, clozapine, diclofenac and either GSH-EE or GSH as a reactive intermediate conjugating agent. An increase in detection sensitivity was observed for each GSH-EE conjugate and in the case of acetaminophen-GSH-EE the peak area increase was approximately 80-fold. Twelve drug compounds, each having known biotransformation mechanisms, were used to further test the detection capabilities of the assay and establish a concordance to literature data. When GSH was used in the assay, conjugates were detected for 4 out of the 12 test compounds (33%). When GSH-EE was used in the assay, conjugates were detected for 10 out of the 12 test compounds (83%). PMID- 15351055 TI - Simultaneous determination of cefepime and grepafloxacin in human urine by high performance liquid chromatography. AB - A liquid chromatographic method with UV detection for simultaneous determination of cefepime and grepafloxacin has been developed. The method uses a C18 column, equipped with a pre-column of the same material, and acetonitrile-0.1 M phosphoric acid/sodium hydroxide buffer (pH 3.0)-0.01 M n-octylamine (pH 3.0) as mobile phase in gradient mode. Mobile flow rate and sample volume injected were 1.3 mL min(-1) and 20 microL, respectively. Detection wavelengths were 259 nm for cefepime and 278 nm for grepafloxacin. The retention times were 4.03 min for cefepime and 8.85 min for grepafloxacin, with detection limits of 1.0 and 1.1 microg mL(-1), respectively. The method was applied to the determination of both antibiotics in spiked samples of human urine. PMID- 15351056 TI - Rapid analysis of docetaxel in human plasma by tandem mass spectrometry with on line sample extraction. AB - A simple, rapid, and sensitive analytical method for the measurement of docetaxel in human plasma was developed and validated. The method is based on positive electrospray ionization tandem mass spectrometry (ESI+-MS-MS) with on-line sample extraction. It uses paclitaxel as internal standard for calibration. The on-line sample extraction minimizes sample handling and is readily adopted for automation. Quantitation of plasma docetaxel was done by the multiple reaction monitoring (MRM) mode. The method had a linear calibration range of 1.00-3000 ng/mL with a correlation coefficient >0.9999. The limit of quantitation (LOQ) for docetaxel in plasma was 1.00 ng/mL. The on-line extraction recovery of docetaxel was between 86.1-94.7%, with %CV < or = 6.1%. This method has high accuracy (90.1 96.3%), and excellent intra-assay (0.6-3.8%) and inter-assay (2.0-5.7%) precision. Its applicability to clinical samples was demonstrated by measuring patient plasma samples after treatment of weekly docetaxel at 25 mg/m2 as 60-min infusion. PMID- 15351057 TI - Development and validation of a high-performance liquid chromatographic method for the determination of cyproterone acetate in human skin. AB - In the framework of a preliminary study on the transdermal penetration of cyproterone acetate (CPA), a simple and rapid procedure involving an extraction step coupled to a HPLC-UV determination has been developed for the separation and quantification of CPA in the two main skin layers-epidermis and dermis-after local application. The separation of epidermis and dermis layers was carefully carried out by means of a sharp spatula after skin immersion in heated water at 65 degrees C. The two skin layers were then treated separately according to the same process: (1) sample homogenization by vibration after freezing with liquid nitrogen in a Mikro-Dismembrator; (2) CPA extraction with methanol after addition of the internal standard (betamethasone dipropionate); (3) centrifugation; (4) evaporation of a supernatant aliquot; (5) dissolution of the dry residue in methanol and addition of water; (6) centrifugation; (7) injection of a supernatant aliquot into the HPLC system. The separation was achieved on octadecylsilica stationary phase using a mobile phase consisting in a mixture of acetonitrile and water (40:60 (v/v)). The method was then validated using a new approach based on accuracy profiles over a CPA concentration range from 33 to 667 ng/ml for each skin layer. Finally, the method was successfully applied to the determination of CPA to several skin samples after topical application of different gel formulations containing CPA. PMID- 15351058 TI - Determination of lansoprazole in pharmaceutical capsules by flow injection analysis using UV-detection. AB - The direct determination of lansoprazole by using a flow injection analysis (FIA) with UV-detection and its application to the pharmaceutical capsules is described, in this study. The best carrier solvent was found to be 0.01 M NaOH and it was determined at optimum conditions such as flow rate of 1 ml min(-1) and wavelength of 292 nm. Examining the repeatability of the method that was found to be 1.72% for intra-day and 2.13% for inter-day precision using the 8.01 x 10(-6) M lansoprazole concentration has validated the method. The linear range of the method was 5.4 x 10(-6) to 5.4 x 10(-5) M. The limit of detection and quantification was found to be 5.8 x 10(-7) and 1.7 x 10(-6) M, respectively. The proposed method was applied to the pharmaceutical capsules and very good results obtained. Thus, the FIA method for the quantification of lansoprazole can be proposed as a cheap, rapid, easy, accurate, and precise method for the routine determination in pharmaceutical preparations. PMID- 15351059 TI - Adsorptive stripping voltammetric quantification of the antipsychotic drug clozapine in bulk form, pharmaceutical formulation and human serum at a mercury electrode. AB - Using the cyclic voltammetry technique and the Britton-Robinson buffer (pH 2-10) as a supporting electrolyte, clozapine was found to reduce at the hanging mercury drop electrode in a single two-electron irreversible step corresponding to reduction of the azomethine group of the seven-member heterocyclic ring. Based on the interfacial adsorptive character of clozapine onto the hanging mercury drop electrode, a validated square-wave adsorptive cathodic stripping (SWAdCS) voltammetric procedure was described for the quantification of bulk clozapine with limits of detection and quantitation of 4.5 x 10(-10) and 1.5 x 10(-9) M, respectively. The proposed procedure was successfully applied to the quantification of the drug in pharmaceutical formulation (Leponex) and human serum without the necessity for samples' pretreatment or any time-consuming extraction or evaporation steps prior to the analysis. The limits of detection and quantitation of clozapine in spiked human serum were found to be 1 x 10(-9) and 3.3 x 10(-9) M, respectively. The proposed procedure for quantification of clozapine in bulk form, tablets and human serum has the advantage of being simple, rapid, sensitive, precise and inexpensive compared to most of the reported methods. PMID- 15351060 TI - Polarographic behaviour of Aceclofenac, Tenoxicam and Droxicam in a methanol water mixture. AB - A polarographic study about how three anti-inflammatories, such as Aceclofenac, Tenoxicam and Droxicam behave, using tast polarography (TP) and differential pulse polarography (DPP) was carried out. These studies were always carried out in a media formed by Methanol-Britton-Robinson aqueous buffer (0.1M) (4:96 (v/v)) due to the low solubility of these drugs in water. A strong influence of pH on analytical signals was observed, showing that the optimal pH values were between 4 and 5. Using DPP in the optimal experimental conditions, a detection limit of 10 ppb for Tenoxicam and Droxicam and 52 ppb for Aceclofenac was reached. The DPP proposed method was successfully applied to the determination of the active compounds in commercial drugs. PMID- 15351061 TI - Simple and rapid HPLC method for determination of amlodipine in human serum with fluorescence detection and its use in pharmacokinetic studies. AB - A fast, sensitive and specific high performance liquid chromatographic method using fluorescence detection is described for analysis of amlodipine in human serum. Amlodipine is extracted from serum by ethyl acetate and involves precolumn derivatization with 4-chloro-7-nitrobenzofurazan (NBD-Cl) and reverse-phase chromatography on C18 column. The mobile phase was sodium phosphate buffer (pH 2.5) containing 1 ml/l triethylamine and methanol at flow rate of 2.8 ml/min. Propranolol was used as internal standard. The standard curve was linear over the range 0.25-16 ng/ml of amlodipine in human serum. The within-day and between-day precision studies showed good reproducibility with coefficients of variation less than 12% for all the analytes. The limit of quantification was 0.25 ng/ml of serum. The method has been applied to a bioequivalence study after administration of 10 mg amlodipine in 12 normal subjects. PMID- 15351062 TI - Determination of nateglinide in animal plasma by micellar electrokinetic chromatography and on-line sweeping technique. AB - A micellar electrokinetic chromatography (MEKC) method was developed for the determination of nateglinide in animal plasma by on-line sweeping technique, in which plasma samples were simply deproteinized with acetonitrile, and analyzed with 16 mmol/L NaH2PO4 + 6 mmol/L Na2B4O7 + 60 mmol/L sodium dodecyl sulfate (SDS) (pH 7.14) as the running buffer, a fused-silica capillary as the separation tube, 21 kV as the running voltage and UV detection at 214 nm. Under these conditions, more than 100-fold enrichment of nateglinide was obtained with the good linear relation in the range of nateglinide plasma concentration 0.2-7 mg/L (R = 0.998). The method could be applied successfully to determine trace drugs in clinical analysis. PMID- 15351063 TI - A validated solid-phase extraction HPLC method for the simultaneous determination of the citrus flavanone aglycones hesperetin and naringenin in urine. AB - A simple, specific, precise, accurate, and robust HPLC assay for the simultaneous analysis of hesperetin and naringenin in human urine was developed and validated. Urine samples were incubated with beta-glucuronidase/sulphatase and the analytes were isolated by solid-phase extraction using C18 cartridges and separated on a C8 reversed phase column using a mixture of methanol/water/acetic acid (40:58:2, v/v/v) at 45 degrees C. The method was found to be linear in the 50-1200 ng/ml concentration range for both hesperetin and naringenin (r > 0.999). The accuracy of the method was greater than 94.8%, while the intra- and inter-day precision for hesperetin was better than 4.9 and 8.2%, respectively and for naringenin was better than 5.3 and 7.8%, respectively. Recovery for hesperetin, naringenin and internal standard 7-ethoxycoumarin was greater than 70.9%. The method has been applied for the determination of hesperetin and naringenin in urine samples obtained from a male volunteer following a single 300 mg oral dose of each of the corresponding flavanone glycosides hesperidin and naringin. The intra- and inter day reproducibility through enzyme hydrolysis was less than 3.9% for both total (free + conjugated) hesperetin and naringenin. Stability studies showed urine quality control samples to be stable for both hesperetin and naringenin through three freeze-thaw cycles and at room temperature for 24 h (error < or = 3.6%). PMID- 15351064 TI - Rapid quantitative analysis of oxiracetam in human plasma by liquid chromatography/electrospray tandem mass spectrometry. AB - A rapid and accurate reversed-phase liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantitative determination of oxiracetam in human plasma. After addition of internal standard (piracetam) plasma was precipitated with two volumes of acetonitrile and the supernatant was evaporated. The residues were dissolved in 0.1% acetic acid and analyzed by reversed-phase HPLC with the detection of the analyte in the multiple reaction monitoring (MRM) mode. This method for the determination of oxiracetam was accurate and reproducible, with a limit of quantitation of 0.2 microg/ml in human plasma. The standard calibration curve for oxiracetam was linear (r2 = 0.999) over the concentration range 0.2-40.0 microg/ml in human plasma. The intra- and inter-day precision over the concentration range of oxiracetam was lower than 8.3% (relative standard deviation, %R.S.D.), and accuracy was between 92.5-106.4%. PMID- 15351065 TI - Automated 96-well liquid-liquid back extraction liquid chromatography-tandem mass spectrometry method for the determination of ABT-202 in human plasma. AB - A high-throughput bioanalytical method using automated sample transferring, automated liquid-liquid back extraction and liquid chromatography-tandem mass spectrometry was developed in a GLP regulated environment for the determination of ABT-202 in human plasma. Samples of 0.30 ml were transferred into 96-well plate using an automatic liquid handler. Automated liquid-liquid extraction (LLE) was carried out on a 96-channel programmable liquid handling workstation using methyl tert-butyl ether as the extraction solvent. A dual-HPLC with single mass spectrometer configuration was utilized to provide a reliable and routine means to increase sample throughput. The standard curve range was 0.38-95.02 ng/ml. There was no interference from endogenous components in the blank plasma tested. The accuracy (% bias) at the lower limit of quantitation (LLOQ) was 7.7% and the precision (% CV) for samples at the LLOQ was 4.7%. The inter-day % CV and % bias of the quality control samples were < or = 6.8 and < or = 7.6%, respectively. Coefficients of determination, a measure of linearity, ranged from 0.994 to 0.997. The method was accurate and reproducible and was successfully applied to generate plasma concentration-time profiles for human subjects after low oral doses of the compound. PMID- 15351066 TI - Ion-pair complex-based solvent extraction combined with chemiluminescence determination of chlorpromazine hydrochloride with luminol in reverse micelles. AB - A new chemiluminescence (CL) method is proposed for the determination of chlorpromazine hydrochloride, which is based on the dichloromethane solvent extraction of ion-pair complex of tetrachloroaurate(III) with chlorpromazine hydrochloride and luminol chemiluminescence detection in a reversed micellar medium formed by the cation surfactant cetyltrimethylammonium bromide in a dichloromethane-cyclohexane (1:1 V/V)-water (0.3 mol/L Na2CO3 buffer solution with the pH of 11.5). The ion-pair complex of tetrachloroaurate(III) with chlorpromazine hydrochloride produced an analytical chemiluminescence signal when it entered the reversed micellar water pool. In the optimum conditions, CL intensities are proportional to concentrations of the studied drug over the range 0.05 approximately 10 microg/mL with a detection limit (DL) of 6 ng/mL. The relative standard deviation (R.S.D.) is 2.6% for 1.25 microg/mL chlorpromazine hydrochloride (n = 11). R.S.D. (precision) of inter-day and intra-day is less than 6%, and accuracy of inter-day and intra-day is satisfactory. The method has been applied to the determination of studied drug in pharmaceutical preparations and biological fluids with satisfactory results. PMID- 15351067 TI - HPLC study of glimepiride under hydrolytic stress conditions. AB - Glimepiride is a modern hypoglycaemic agent, which belongs to the group of sulfonylurea derivates. In this paper, simple, specific and accurate RP-HPLC method was developed in order to study decomposition of glimepiride under the hydrolytic stress conditions (acid, neutral, alkaline and oxidative). The best separation of glimepiride and its degradation products was achieved on reverse phase C18 column. The mobile phase was composed of acetonitrile-phosphate buffer (pH 3.5, 0.03 M) (48:52, v/v). Employing RP-HPLC method, five main degradation products were detected in the exposed samples. It was found that the susceptibility of glimepiride to hydrolytic decomposition increased in following manner: neutral condition < alkaline condition < acid condition < oxidative condition. PMID- 15351068 TI - Analysis of fluorescence excitation-emission matrix of multicomponent drugs. A case study with human placental extract used as wound healer. AB - The possibility of finger printing of multi component drugs, where a number of components are fluorescent, has been explored after generation of excitation emission matrix (EEM) contour plots. With nicotinamide adenine dinucleotide, reduced form (NADH) and N-acetyl tryptophanamide (NATA) serving as reference compounds, the contour widths were found to be proportional to fluorophore concentration. EEM generated from different batches of processed human placental extract, used as wound healer, in presence and absence of extraneous fluorescence probes like 1-anilino 8-naphthalenesulphonic acid (1-ANS) or dansyl chloride were found to be highly consistent. Variation of contour widths of these EEM after excitation at 280 or 340 nm were essentially zero subject to the limit of detection of variation (n = 5). This high degree of consistency between batches has been supported from analysis with such independent techniques like UV absorption spectra, Fourier Transformed IR spectra, TLC and reverse phase HPLC. The advantages of EEM analysis have been discussed. PMID- 15351069 TI - Determination of dopamine by flow-injection analysis coupled with luminol hexacyanoferrate(III) chemiluminescence detection. AB - A novel flow-injection method (FIA) for the determination of dopamine based on the inhibition of the intensity of chemiluminescence (CL) from luminol hexacyanoferrate(III) system in basic medium is described. The present method allows the determination of dopamine over the range 30-100 microg l(-1) and 400 3000 microg l(-1). The relative standard deviation is 2.32% for 70 microg l(-1) dopamine and 1.22% for 1500 microg l(-1) dopamine (n = 20). The detection limit is 5 microg l(-1) with the sampling rate of 135 samples h(-1). This method has been applied for the determination of dopamine in commercial pharmaceutical injection samples. The results obtained by this method agreed with those by the official method. PMID- 15351070 TI - An expression of within-plate uncertainty in sandwich ELISA. AB - This paper puts forward a method to describe an equation of the within-plate uncertainty (relative standard deviation (R.S.D.) of measurements) as a function of analyte concentration in sandwich enzyme-linked immunosorbent assay (ELISA). A kit for thyroid stimulating hormone is taken as an example. The pipetting procedures of analyte solution and chromogen-substrate solution and absorbance inherent to the wells of a microplate are identified as major error sources and their variability is included as parameters in the uncertainty equation. These parameters can be determined by the experiments with distilled water. The theoretical R.S.D. is shown to be in good agreement with the results of the repeated experiments using the real samples. Since the theory gives a continuous plot of R.S.D. against concentration, the uncertainty structure of the ELISA kit can be recognized over a wide concentration range and the detection limit and quantitation range can easily be determined on the plot. PMID- 15351071 TI - Identification and characterization of major degradation products of risperidone in bulk drug and pharmaceutical dosage forms. AB - Acid, base and oxygen stability of risperidone, a novel anti-psychotic drug, has been evaluated storing the sample in solution phase. One of the major degradation products has been identified and characterized by using techniques namely IR, MS and NMR after isolation by preparative LC. The other major degradation product has been identified with help of MS/MS data and by co-eluting in analytical LC with the available standard. The effect of acid and base resulted in the formation of hydroxy risperidone and the effect of oxygen lead to the formation of N-oxide of risperidone. The two major degradation products in the dosage forms were also characterized as 9-hydroxy risperidone and N-oxide of risperidone, after enrichment through preparative LC, by LC-MS/MS and HPLC. Structural elucidation of degradation product leading to the formation of N-oxide of risperidone is discussed in detail. PMID- 15351072 TI - Chemiluminescence detection of isoniazid using Ru(phen)3(2+)-isoniazid-Ce(IV) system. AB - In our experiment, it was observed that isoniazid could enhance the chemiluminescence (CL) emission of tris-(1,10-phenanthroline)ruthenium(II) (Ru(phen)3(2+))-cerium(IV) (Ce(IV)) system and this enhancement effect was dependent on the concentration of isoniazid, based on which, a novel CL system was established for the detection of isoniazid. Under the optimum experimental conditions, the dynamic range and detection limit are 7.0 x 10(-2) to 6.5 microg ml(-1) and 2.5 x 10(-2) microg ml(-1), respectively. The R.S.D. is 3.4% (n = 11). The proposed method has been applied to detect the content of isoniazid in the injection solution with satisfactory results. The possible mechanism of the CL reaction was studied. PMID- 15351073 TI - HPLC method for simultaneous determination of cefprozil diastereomers in human plasma. AB - A high-performance liquid chromatography method was developed for the determination of cefprozil diastereomers in human plasma. Cefprozil exists as cis and trans isomer at the ratio of 90:10. Plasma samples were prepared by protein precipitation using acetonitrile, trichloroacetic acid and methylene chloride. After the mixtures were vortexed and centrifuged, the aqueous supernatant was injected into a reversed-phase C8 column. The mobile phase consisted of acetonitrile, glacial acetic acid and distilled water at the volume ratio of 5.5:1.75:92.75 (pH 2.7). The signals were monitored with UV detection at 280 nm. The calibration curves of cis and trans isomer were linear in concentration ranges of 0.1-25 and 0.02-2.5 microg/mL with the correlation coefficient of 0.9999 and 0.9989, respectively. After oral administration of cefprozil in humans, Cmax and Tmax of total cefprozil were 18.80 +/- 2.14 microg/mL and 2.06 +/- 0.62 h. This method was sensitive with excellent selectivity and reproducibility, and successfully applied to a bioavailability study of cefprozil in healthy subjects. PMID- 15351074 TI - Founders of child neurology in Japan--Masaki Suzuki. PMID- 15351076 TI - Developing the brain -- a proposal to pediatricians. AB - What we should do as pediatric neurologists to solve educational problems may be not only to find symptomatic solutions but also to drastically re-evaluate 'the view of children'. Brain science, rapidly advancing at present, gives us some new scientific achievements, such as neuronal cell death, synaptic overproduction/elimination, sensitive period, higher cortical function of neonates, and neural correlates, useful for this re-evaluation. From this aspect, I have made a proposal for the establishment of new developmental neurology. PMID- 15351077 TI - 'Nurturing the brain' as an emerging research field involving child neurology. AB - 'Nurturing the brain' is an emerging research field integrating brain science, child care and education, which also involves child neurology. It has emerged from the recent remarkable progress in brain science and strong social demands for improvements in child care and education. This article reviews the current status of three major research themes in this field. First, developmental disorders represented by attention deficit/hyperactivity disorder, autism and Asperger syndrome often introduce difficulties in child care and education, which are to be addressed by appropriate assessment and treatment of affected children based on new knowledge of the pathogenesis of these disorders. Second, recent progress in research on the critical/sensitive periods of development of brain structures and functions promises useful advice for teachers and parents regarding optimal timing and ways of teaching various subjects. Third, the development of the brain throughout infancy, childhood and adolescence is paralleled by the growth and maturation of the mind. Neuronal mechanisms underlying the theory of mind, mirror neurons, internal model, cognitive control, and cognitive emotion regulation are important themes that bridge our understandings of the brain and the mind. PMID- 15351078 TI - The concept of 'developing the brain': a new natural science for learning and education. AB - From the viewpoint of biology, learning and education can be defined as the processes of forming neuronal connections in response to external environmental stimuli, and of controlling or adding appropriate stimuli, respectively. Learning and education can thus be studied as a new field of natural sciences with the entire human life span as its subject, thus including various problems such as fetal environment, childcare, language acquisition, general/special education, and rehabilitation. Non-invasive imaging of higher-order brain functions in humans will clarify the brain's developmental processes, and will provide various evidence for learning sciences. This new approach is called 'developing the brain' or 'brain science and education'. The origin of the concept and its present state are described and its future prospects are briefly analyzed. PMID- 15351079 TI - Learning and memorization impairment in childhood chronic fatigue syndrome manifesting as school phobia in Japan. AB - For the last 15 years, we have tried to understand the pathophysiology of childhood chronic fatigue syndrome (CCFS) in Japan. In this condition, two major symptoms are important: easy fatigability and disturbed learning and memorization. In CCFS patients we clinically evaluated autonomic nervous system function, circadian rhythm of hormonal secretion (melatonin, cortisol and 3 endorphin), core body temperature, and sleep-wake pattern. Most patients showed autonomic nervous system dysfunction and circadian rhythm disturbances, similar to those observed in jet lag. Radiological imaging studies (SPECT, Xe-CT, and MRS) revealed decreased blood flow in the frontal and thalamic areas, and accumulation of choline in the frontal lobe. We analyzed the relationship between the laboratory data and clinical symptoms in CCFS. PMID- 15351080 TI - Theophylline impairs memory/learning in developing mice. AB - We studied the relationship between theophylline and memory/learning using an elevated plus-maze test and measuring spontaneous locomotor activity in developing mice. There were no significant differences in transfer latency (TL) in 21-, 30- and 42-day-old mice in the acquisition trial, but theophylline significantly prolonged TL in comparison with the control group in 21- and 30-day old mice in the retention trial (n = 20; 21-day-old mice, P = 0.0009; 30-day-old mice, P = 0.03). The dose-dependent stimulant effects of theophylline were shown in spontaneous locomotor activity (n = 16; 21-day-old mice, P = 0.0005; 30-day old mice, P = 0.0007; 42-day-old mice, P = 0.0003) in all groups and in rearing in 21-day-old mice (P = 0.02) 1 h after drug administration, but there were no significant effects 24 h after drug administration in all groups. Our study showed that even a therapeutic dose of theophylline impaired memory/learning only in developing mice, and theophylline increased locomotor activity both in developing and adolescent mice. These observations are consistent with several clinical studies, which reported the relationships between theophylline use and memory/learning and learning-related behaviors in children, such as difficulty in studying and hyperactivity. Theophylline possibly induces memory/learning disabilities and hyperactivity in infants and young children. PMID- 15351081 TI - Dichloroacetate treatment for mitochondrial cytopathy: long-term effects in MELAS. AB - The long-term effects of the sodium salt of dichloroacetic acid (DCA) were evaluated in four patients with mitochondrial encephalomyelopathy with lactic acidosis and stroke-like episodes (MELAS) carrying A3243G mutation. Oral administration of DCA in MELAS patients was followed for an average of 5 years 4 months. Serum levels of lactate and pyruvate were maintained at around 10 and 0.6 mg/dl, respectively. Serum levels of DCA were 40-136 microg/ml. Symptoms responding to treatment included persistent headache, abdominal pain, muscle weakness, and stroke-like episodes. In contrast, no improvements in mental status, deafness, short stature, or neuroelectrophysiological findings were observed. Adverse effects included mild liver dysfunction in all patients, hypocalcemia in three and peripheral neuropathy in one. None of these adverse events was severe enough to require discontinuation of treatment. To determine suitable indications for DCA therapy, analysis of many more patients who have undergone DCA administration is required. PMID- 15351082 TI - Clinical features of A3243G mitochondrial tRNA mutation. AB - Mitochondrial cytopathy is a heterogeneous group of disorders with a wide range of clinical features. To evaluate the incidence and clinical heterogeneity of A3243G mitochondrial tRNA mutation in the Korean population, we evaluated patients who were clinically suggestive of having mitochondrial encephalomyopathy. Eighty-five patients were included in this study. All showed clinical features of mitochondrial encephalomyopathy and had three or more of the following clinical manifestations: (1) psychomotor regression, (2) hyperlacticacidemia, (3) recurrent stoke-like episodes, (4) idiopathic cardiomyopathy, (5) sensoryneural hearing loss, (6) diabetes mellitus, (7) myopathy, (8) renal disease and (9) relatives with known mitochondrial disease. The patients were clinically classified as MELAS, MERRF, Leigh syndrome, Kearns Sayre syndrome, chronic progressive external ophthalmoplegia and uncertain. Of the 85 patients, 19 had the A3243G mutation (22.3%). Thirty-one patients showed typical clinical characteristics of MELAS. Fourteen of those 31 patients had A3243G mutation (45.1%). Four patients harboring A3243G mutations showed atypical and heterogeneous clinical features, unlike MELAS. This study revealed the frequent occurrence of A3243G mutation in Korean patients with mitochondrial disorders and their clinical features can be heterogeneous. It will be helpful to screen the presence of A3243G mutation for the genetic diagnosis of mitochondrial encephalomyopathy in Korea. PMID- 15351083 TI - Gait control in spinal palsy. AB - Developmental motor impairment with lower limb spasticity most commonly corresponds to cerebral palsy of the spastic diplegia type. Here we describe a 4 year-old girl whose locomotor phenotype reflects early cortico-spinal lesion at the spinal level. This child has developmental spastic paraparesis secondary to D4-D8 cord compression. We analysed her gait using the ELITE optoelectronic system and compared it to that of six normal age-matched controls and six age matched children with leucomalacic spastic diplegia. Gait characteristics of the patient included preservation of head orientation and arm swing similar to findings in normal controls and contrasting with children with spastic diplegia. She also had truncal instability and displayed lack of selectivity in lower limb movement as in spastic diplegia and in contrast with normal controls. This may reflect differences in locomotor control between developmental spasticity of cerebral and spinal origin. The latter might correspond to spinal palsy defined as abnormal movement and posture secondary to non-progressive pathological processes affecting the immature spinal cord. PMID- 15351084 TI - Fukutin expression in mouse non-muscle somatic organs: its relationship to the hypoglycosylation of alpha-dystroglycan in Fukuyama-type congenital muscular dystrophy. AB - Recent studies suggest that hypoglycosylation of alpha-dystroglycan (alpha-DG) may play an essential role in the pathogenesis of Fukuyama-type congenital muscular dystrophy (FCMD), which is caused by defects in the fukutin gene and characterized by dystrophic changes in the skeletal muscles and dysplastic lesions in the central nervous system. alpha-DG is expressed in many organs in addition to muscle and brain, although these organs are not affected in FCMD. To elucidate whether or not fukutin protein is involved in the glycosylation of alpha-DG in non-muscle somatic organs, we examined the distribution pattern of fukutin in developing and adult mouse tissues. The fukutin antisera labeled the acinar cells of the pancreas, the renal glomerular and tubular cells, and the epithelium of the bronchi, salivary gland, alimentary tract and skin in both fetal and adult mice. This distribution pattern was also confirmed by in situ hybridization. Antisera against alpha-DG and beta-DG labeled the same cellular populations in each organ, especially along the cell surface membrane. We also examined the glycosylation status of alpha-DG in autopsied FCMD cases (n = 5) and found evidence of hypoglycosylation in the kidney, lung, skin and intestine. These results suggest that fukutin protein is involved in the glycosylation process of alpha-DG in non-muscle somatic organs both during development and in the adult. It is unclear why muscle and brain symptoms predominate in FCMD, however re-evaluation of the functions of alpha-DG and fukutin protein in non muscle somatic organs may provide valuable insight. PMID- 15351086 TI - Beneficial effect of L-arginine for stroke-like episode in MELAS. AB - We here reported the clinical course and therapeutic details of a 16-year-old girl with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke like episodes (MELAS) who had had five stroke-like episodes (two episodes were clinically mild, while the three subsequent episodes were severe). Among the three episodes, the symptoms improved earliest and magnetic resonance spectroscopy abnormality was minimal when given L-arginine in addition to prednisolone, glycerol and edalavone. L-arginine administration during the acute phase of MELAS might be a potential therapy to reduce brain damage due to mitochondrial dysfunction. PMID- 15351087 TI - Systemic growth hormone corrects sleep disturbance in Smith-Magenis syndrome. AB - Smith-Magenis syndrome (SMS) is a multiple congenital anomaly syndrome characterized by an interstitial deletion of chromosome 17p11.2. Sleep problems such as nocturnal awakening and abnormality in the percentage of rapid eye movement (REM) sleep are frequently observed in patients with SMS, and several medications have been administered to improve the sleep disorders. Here we present a female case of SMS showing early waking and reduction of REM sleep, which were corrected by human growth hormone (GH) replacement for her dwarfism. Also, we report changes in the sleep-wakefulness circadian rhythm and polysomnographical data before and after the start of human GH replacement. It is speculated that GH deficiency could be involved in sleep disturbance in SMS. PMID- 15351088 TI - Non-epileptic pedaling-like movement induced by triclofos. AB - We reported an otherwise healthy infant with non-epileptic pedaling-like movement induced by triclofos. For the purpose of sedation for head computed tomography, 80 mg/kg of triclofos was administered. Fifteen minutes after the use of triclofos, the infant exhibited pedaling-like movement accompanied by slight upper eye deviations. This movement could be arrested by manual restraint. The pedaling-like movement lasted for about 2 h intermittently. Electroencephalography during this movement revealed physiological activities with semirhythmic theta waves without paroxysmal activities. PMID- 15351089 TI - Hot water epilepsy and focal malformation of the parietal cortex development. AB - Hot water epilepsy (HWE) refers to a specific type of reflex epilepsy precipitated by the stimulus of bathing in hot water. HWE is considered to be a geographically specific epileptic syndrome since it mainly occurs in the Indian community. Spontaneous seizures may also occur later in life. The seizure pattern includes complex partial attacks. Although the pathogenesis of HWE is still unknown, temporal lobe has been thought to take part in the epileptogenesis. This paper reports on a 4-year-old girl who, at the age of 6 months, experienced complex partial seizures triggered by bathing in hot water. Non-provoked seizures intercritical EEG showed isolated spikes and spike-and-waves in the left parietal region. Brain MRI detected a left parietal focal cortical dysplasia. This is the second patient with HWE in whom a cortical malformation has been observed. The observation present here and data reported in the literature seem to indicate that the sensory cortex might also be involved in triggering seizures precipitated by a bath in hot water. Moreover, the authors believe that MRI examination should be considered for this group of patients. PMID- 15351091 TI - Development of the human breast. AB - The human breast undergoes a complete series of changes from intrauterine life to senescence. These changes can be divided into two distinct phases; the developmental phase and the differentiation phase. The developmental phase includes the early stages of gland morphogenesis, from nipple epithelium to lobule formation. In lobule formation, both processes, development and differentiation, take place almost simultaneously. For example, the progressive transition of lobule type 1 to types 2, 3, and 4 requires active cell proliferation, to acquire the cell mass necessary for the function of milk secretion. This later process implies differentiation of the mammary epithelium. Therefore, the presence of lobule type 4 is the maximal expression of development and differentiation in the adult gland, whereas the presence of lobule type 3 could indicate that the gland has already been developed. It is important to point out that the presence of proteins that are indicative of milk secretion, such as alpha-lactalbumin, casein, or milk fat lobule type membrane protein, also indicates cellular differentiation of breast epithelium. However, only when all the other components of milk, (such as lactose, alpha-lactalbumin, casein and milk fat) are coordinately synthesized within the appropriate structure can full differentiation of the mammary gland be acknowledged. PMID- 15351092 TI - Steroidal hormones and proliferation, differentiation and apoptosis in breast cells. AB - The impact of estrogens (E) and progestins (P) on the breast is crucial. Recent epidemiological studies raised a great concern concerning breast cancer risk and hormone replacement therapy (HRT). However, the effects of HRT in breast tissue remain unclear. Biological data predominantly show that P are antiproliferative and proapoptotic at least for normal breast cells. These antiproliferative effects of P are well described at the cellular level. Whereas E2 increases the level of the various cyclins involved in the cell cycle progression and decreases the cyclin kinase inhibitors, p21 and p27, progestins act in an opposite manner. In addition, they both modulate the phosphorylated rate of Rb involved into the S phase progression. Various proteins of the apoptotic cascade are also targets for E2 and P. We showed that bcl-2, p53 and caspase 3 are oppositely modulated by E2 and P in normal and breast cancer cell cultures. It is very possible that in vivo the balance between E2/P, the type of P, specific phenotypes could explain increasing risk during HRT, which appears to be mainly a promoter effect on preexisting transformed cells. PMID- 15351093 TI - Local biosynthesis and metabolism of oestrogens in the human breast. AB - The origin of oestrogens at the level of the breast itself is discussed. In particular in postmenopausal women an accumulation of oestradiol at the site of breast tumours has been documented by a number of independent studies. The mechanism behind the high local oestrogens concentrations is thought to be the in situ production of these steroids by local processes with androstenedione as the main precursor. The presence of all enzymes required for this production has been demonstrated in a large proportion of breast tumours, with probably aromatase, hydroxysteroid dehydrogenase type 1 and sulfatase as the most important enzymes leading to the biologically highly active oestradiol. The individual enzymes that are relevant for the biosynthesis and the metabolism of oestrogens are discussed. The conclusion is reached that a number of these local processes may be involved in the promotion of premalignant lesions and in stimulation of growth of malignant tumours in the human breast. PMID- 15351094 TI - Oncogenic pathways in hereditary and sporadic breast cancer. AB - Cancer is a genetic disease. Breast cancer tumorigenesis can be described as a multi-step process in which each step is thought to correlate with one or more distinct mutations in major regulatory genes. The question addressed is how far a multi-step progression model for sporadic breast cancer would differ from that for hereditary breast cancer. Hereditary breast cancer is characterized by an inherited susceptibility to breast cancer on basis of an identified germline mutation in one allele of a high penetrance susceptibility gene (such as BRCA1, BRCA2, CHEK 2, TP53 or PTEN). Inactivation of the second allele of these tumour suppressor genes would be an early event in this oncogenic pathway (Knudson's "two-hit" model). Sporadic breast cancers result from a serial stepwise accumulation of acquired and uncorrected mutations in somatic genes, without any germline mutation playing a role. Mutational activation of oncogenes, often coupled with non-mutational inactivation of tumour suppressor genes, is probably an early event in sporadic tumours, followed by more, independent mutations in at least four or five other genes, the chronological order of which is likely less important. Oncogenes that have been reported to play an early role in sporadic breast cancer are MYC, CCND1 (Cyclin D1) and ERBB2 (HER2/neu). In sporadic breast cancer, mutational inactivation of BRCA1/2 is rare, as inactivation requires both gene copies to be mutated or totally deleted. However, non-mutational functional suppression could result from various mechanisms, such as hypermethylation of the BRCA1 promoter or binding of BRCA2 by EMSY. In sporadic breast tumorigenesis, at least three different pathway-specific mechanisms of tumour progression are recognizable, with breast carcinogenesis being different in ductal versus lobular carcinoma, and in well differentiated versus poorly differentiated ductal cancers. Thus, different breast cancer pathways emerge early in the process of carcinogenesis, ultimately leading to clinically different tumour types. As mutations acquired early during tumorigenesis will be present in all later stages, large-scale gene expression profiling using DNA microarray analysis techniques can help to classify breast cancers into clinically relevant subtypes. PMID- 15351095 TI - Postmenopausal hormone therapy and the risk of breast cancer: the view of an epidemiologist. AB - OBJECTIVES: Postmenopausal hormone therapy (PMH) has been widely used by menopausal women living in western countries for the past several decades. Numerous studies have evaluated the relationship between PMH and breast cancer risk because steroid hormones have been implicated in breast cancer etiology. METHODS: A review of selected studies was performed to evaluate the history of investigations of the association between PMH and breast cancer, with a focus on studies evaluating different PMH regimens and different histologic types of breast cancer. RESULTS: Though studies conducted before the early 1990s suggest that both combined estrogen and progestin (E + P) PMH and unopposed estrogen (E) PMH are associated with an increased risk of breast cancer, more recent observational studies suggest that E + P, particularly current use for 5 years or longer, is more strongly associated with breast cancer risk than is unopposed E. Results from the Women's Health Initiative (WHI) randomized trials have confirmed these findings as they indicate that E + P is causally related to breast cancer (relative risk (RR) = 1.24; 95% confidence interval (CI): 1.01-1.54), while E alone is not (RR = 0.77; 95% CI: 0.59-1.01). CONCLUSIONS: There is clear and consistent evidence that use of E + P increases a woman's risk of breast cancer. Alternatively, current evidence suggests that use of unopposed E is not as strongly associated with breast cancer risk. Further studies are needed though to examine how different PMH regimens, doses, and methods of delivery are related to breast cancer risk, and how PMH impacts the risks of different types of breast cancer. PMID- 15351096 TI - Postmenopausal hormone therapy and the risk of breast cancer. A clinician's view. AB - Reports from the Women's Health Initiative (WHI) and the Million Women Study have indicated that postmenopausal hormone therapy increases the risk of breast cancer. At this point in time, it is not certain whether these data reflect a small increase in risk or an impact of hormone therapy on pre-existing tumors. The purpose of this review is to provide an analysis of the epidemiologic data that can help the clinician inform patients and assist patients in their decision making. PMID- 15351097 TI - Exogenous progestagens and the human breast. AB - The role of progestins (or progestagens) on the breast tissue remains controversial. However, according to the molecule and the duration of application, cell differentiation and apoptosis may predominate over proliferation. Progestins are also used as second-line agents for the treatment of metastatic breast cancer. In young women with benign breast disease, long-term treatment with 19-nortestosterone progestins had a trend to decrease breast cancer risk contrarily to what was observed in postmenopausal women receiving estrogens. Several compounds with progestational activity have been used for HRT. Small differences in the structure of the molecules may lead to pronounced differences in activities, some progestins exerting androgenic effects and some exerting estrogenic or glucocorticoid like activities. While most progestins do not bind to the estrogen receptors, it has been shown that some androgenic progestins stimulate MCF7 cells proliferation while progestins derived from progesterone did not induce cell multiplication in the same cell lines. Therefore, different progestins may induce different effects on the breast cells. Whether the progestins available to date are able to bind specifically to the progesterone receptors PR-A or PR-B and whether this is of clinical relevance to breast cell proliferation is still unclear. Although the relationship between progestin use and breast cancer risk is still the subject of debate and controversy, the data reported to date suggest that 5 years of treatment carry a low risk but further duration of use increases the risk. Further studies are still needed, randomised long-term prospective studies as well as from the laboratory, especially to determine whether a sequential or continuous regimen would be preferable as far as breast-cell response and apoptosis are concerned, and what are the effects of the various molecules used for HRT. PMID- 15351098 TI - Hormones and mammographic breast density. AB - Mammographic density reveals information about the hormonal environment along with the heritability in which breast cancer develops. This is made possible by the widespread use of population screening by mammography. Increasingly this is an important observation not just for population studies, which reveal disease determinants, but also for the individual. Density reveals the effect of the intrinsic hormonal environment and its background genetics, and also the effect of pharmaceuticals--agents used for disease control and prevention and hormone replacement therapy (HRT) used for well-being around the menopause. Increasingly this focus on the individual will need methods of measurement of density that can be monitored with greater accuracy than the widely used BI-RADS 4 categories. For this purpose studies are under way to measure volume of dense tissue as a continuous variable. In due course, measurement of density will be used as a biomarker of risk, employed in risk models and to monitor interventions. Before this can happen more knowledge will be needed of the change occurring naturally through the menopause and the differences between individuals. This will need specific study backed up with detailed information about the patient on large numbers of women and their mammograms. Currently the widespread use of HRT has increased the prevalence of the dense patterns and potentially may adversely affect the effectiveness of mammographic screening programmes. There is a large literature recording this from which we see that combined continuous preparations of oestrogen progestin are more likely to cause increased density than oestrogen alone or tibolone. Breast density, measured more accurately, has the potential to be an important adjunct to risk estimation and to monitor interventions for breast cancer prevention with pharmaceuticals (such as SERMS) and by change in lifestyle behaviours. PMID- 15351099 TI - Lessons to be learned from animal studies on hormones and the breast. AB - The relation of hormone use by postmenopausal women to breast cancer risk has been controversial and unclear. A recent large randomized trial, the Women's Health Initiative (WHI) and a large observational study (Million Women Study) provided somewhat conflicting answers. The WHI found an increased incidence of breast cancer among women given hormone therapy (conjugated equine estrogen plus medroxyprogesterone acetate) but no increase in those given estrogen only therapy (conjugated equine estrogen alone). Whereas, the Million Women Study found an increased breast cancer risk among the estrogen plus progestin and the estrogen only users. This review brings comparative perspective to the issue of the effects of estrogen plus progestin versus estrogen only effects on breast cancer and is focused particularly on nonhuman primates. Although data from rodents is mixed, studies of monkeys suggest that estrogen only treatment has little or no effect on breast cell proliferation, and by inference, on breast cancer risk. On the other hand, data from both mouse and monkey studies strongly support the conclusion that the co-administration of a progestogen with an estrogen markedly increases breast cell proliferation and the potential for breast cancer promotion. PMID- 15351100 TI - Lessons to be learned from clinical studies on hormones and the breast. AB - Estrogen is a well-known mitogen in human breast epithelium but the action of progestogen is complex and incompletely understood. During the last years, accumulating data from animal, clinical and observational studies suggest a proliferative effect in breast tissue when progestogen is added to estrogen. Findings in surrogate markers like breast density add to clinical and epidemiological reports indicating that continuous combined HRT may carry a higher risk of breast cancer than treatment with estrogen alone. Whether the results are valid for all progestogens remains to elucidated. It is also clear that not all women respond in the same way to the same treatment and the biological basis for the marked individual variation in breast response has to be clarified. Further knowledge about the role of androgens and of the impact of different treatment regimens is important and prospective randomized clinical studies are needed. PMID- 15351101 TI - Breast cancer and HRT use. PMID- 15351102 TI - Tissue-selective effects of tibolone on the breast. AB - Hormone treatment with an estrogen plus a progestagen (EPT) increases the risk of breast cancer. Both hormone activities are also induced by tibolone. In order to assess the breast safety of tibolone, it was evaluated in several pre-clinical models. The effects were inconclusive in breast cancer cell lines but, in various in vivo models, it did not stimulate the breast. In the 17,12 dimethylbenz(a)anthracene (DMBA) model, tibolone clearly inhibited the growth of breast tumors and, when given prophylactally, far less tumors developed. Ovariectomized monkeys showed no increase in the expression of the proliferation marker Ki67. The effects of tibolone and its metabolites on the steroid metabolizing enzymes in breast tissues were investigated in order to unravel its mode of action in the breast. Tibolone and its metabolites did not inhibit aromatase, but sulfatase was profoundly inhibited. The sulfated 3alpha-OH tibolone metabolite even showed irreversible inhibition of sulfatase. In addition, 17ss-hydroxysteroid dehydrogenase activities were slightly inhibited and sulfotransferase activity was stimulated at low concentrations. The consequence of these effects is that, for both endogenous estrogens and estrogenic-metabolites of tibolone, the equilibrium is preferential for the sulfated forms. The intracellular hormonal milieu tibolone and its metabolites also influence cellular homeostasis. It inhibits cell proliferation of normal breast epithelial cells and stimulates apoptosis. In this respect, tibolone behaves differently from estrogens. Clinical studies have shown that tibolone users experience less breast tenderness and do not show an increase in mammographic density as found with continuous combined EPT. The data concerning tibolone and breast cancer risk are inconclusive and require further investigation. PMID- 15351103 TI - The effects of tibolone and oestrogen-based HT on breast cell proliferation and mammographic density. AB - Tibolone is a tissue-selective compound used for the treatment of climacteric symptoms and the prevention of osteoporosis in post-menopausal women. In this review some in vitro data and clinical studies indicating that the effects of tibolone on breast tissue are different from those seen with oestrogen-based hormone therapy (HT) are briefly discussed. From a clinical perspective, an increase in mammographic density and breast cell proliferation should be regarded as an unwanted side-effect of HT. Efforts should therefore be made to define treatment regimens for post-menopausal women that have minimal effects on the breast but still maintain the many advantages of HT. Data suggest that tibolone may be such an alternative. PMID- 15351104 TI - Postmenopausal hormone therapy before and after breast cancer: clinical experiences. AB - Conventional oestrogen-based hormone therapy (HT) increases the incidence of breast pain and tenderness, mammographic density and the risk of breast cancer. Combined oestrogen plus progestogen therapy (EPT) increases the risk of breast cancer to a greater degree than oestrogen alone (ET). Attention must therefore be focused on identifying women at risk of breast cancer or on producing a HT that has fewer breast side effects. Randomised controlled trials have shown that while EPT induces breast tenderness or pain in up to 50% of women and increases mammographic density in up to 70% during the first year of treatment, only about as many as one-tenth women report breast tenderness or pain with tibolone and increases in mammographic density are rare, occurring with a similar incidence as seen in untreated controls. Many women with breast cancer suffer vasomotor symptoms rather than risk recurrence with conventional HT. However, in a small randomised controlled trial in women with early breast cancer undergoing adjuvant tamoxifen treatment, tibolone reduced hot flushes, night sweats and improved quality of life compared with placebo. PMID- 15351105 TI - Hypoglycemic activity of Bauhinia candicans in diabetic induced rabbits. AB - The effect of different fractions of methanolic extract of Bauhinia candicans leaves were investigated on experimentally-induced diabetic rabbits. The crude extracts (8 mg/kg) showed hypoglycemic activity along with a reduced urinary glucose excretion. Among the fractions, the highest activity was exhibited by the butanolic fraction (fraction III). Moreover, fraction III reduced plasma glucose level also in normal, as well as, glucose loaded rabbits. These results suggest that B. candicans increases the peripheral metabolism of glucose. PMID- 15351106 TI - A new acylated quercetin glycoside from Ranunculus lanuginosus. AB - A new acylated quercetin glycoside, quercetin 3-O-(2-t-p-coumaroyl)-beta-D glucopyranosyl-(1-->2)-beta-D-glucopyranoside-7-O-beta-D-glucopyranoside (1), together with the known 3-(3,4-dihydroxyphenyl)lactic acid (2) were isolated from Ranunculus lanuginosus leaves and identified on the basis of detailed spectral analysis, including 2D-NMR spectrometry and ESI-MS. PMID- 15351107 TI - Chronic toxicity of Gynostemma pentaphyllum. AB - The effect of water extract of Gynostemma pentaphyllum was evaluated on 6-month chronic toxicity in Wistar rats. Control group received orally 10 ml kg(-1) day( 1). The extract was orally given to the five treatment groups at the doses of 6, 30, 150, 750 and 750 mg kg(-1) day(-1) for 24 weeks. The last group served as the recovery group. The results showed that the extract did not produce any significant dose-related changes. Therefore, it is concluded that the extract of G. pentaphyllum at the given doses did not produce any significant toxic effect in rats during 6-month period of the treatment. PMID- 15351108 TI - A new ent-kaurane diterpenoid from Croton tonkinensis leaves. AB - A new ent-kaurane diterpenoid (1) was isolated from the leaves of Croton tonkinensis. The structure of 1 was determined as ent-7beta-hydroxy-15-oxokaur-16 en-18-ol from spectroscopic methods. PMID- 15351109 TI - Antiinflammatory, analgesic and antioxidant activities of the fruit of Foeniculum vulgare. AB - Oral administration (200 mg/kg) of Foeniculum vulgare fruit methanolic extract exhibited inhibitory effects against acute and subacute inflammatory diseases and type IV allergic reactions and showed a central analgesic effect. Moreover, it significantly increased the plasma superoxide dismutase (SOD) and catalase activities and the high density lipoprotein-cholesterol level. On the contrary, the malondialdehyde (MDA) (as a measure of lipid peroxidation) level was significantly decreased in F. vulgare fruit methanolic extract group compared to the control group (P<0.05). These results seems to support the use of F. vulgare fruit methanolic extract in relieving inflammation. PMID- 15351110 TI - Antifungal limonoids from the fruits of Khaya senegalensis. AB - Investigation of the fruits of Khaya senegalensis resulted in the isolation of three new mexicanolide limonoids containing a rare conjugated diene lactone system named seneganolide A (1), 2-hydroxyseneganolide A (2) and 2 acetoxyseneganolide A (3). Two known limonoids, 3-deacetyl-7-deacetoxy-7 oxokhivorin (4) and methyl 6-hydroxyangolensate (5), were also found. The structures of the new compounds were elucidated on the basis of spectral methods. The antifungal activity of compounds 1, 3 and 5 was tested against the fungus Botrytis cinerea. PMID- 15351111 TI - Germacranolides from Anvillea radiata. AB - The aerial parts of Anvillea radiata yielded a new germacranolide, 8alpha,9alpha epoxyparthenolide (3), together with two known compounds, 9alpha hydroxyparthenolide (1) and parthenolid-9-one (2). The structures of the compounds were elucidated from IR, HRMS, 1H and 13C-NMR, COSY, HETCOR, HMBC and HOHAHA spectral data. The major component 1 was tested for its cytotoxicity and antibacterial activity. PMID- 15351112 TI - Antimicrobial activity and cytotoxicity of Cirsium tenoreanum. AB - The methanol and ethyl acetate extracts of the aerial parts of Cirsium tenoreanum, have been evaluated for their phytochemical constituents and antibacterial and cytotoxic properties. Three flavonoids, apigenin, quercetin-3-O galactoside and kaempferol-3-O-ramnoside, were isolated. The ethyl acetate extract showed a good antiproliferative activity. PMID- 15351113 TI - Antifungal properties of Pseudarthria viscida. AB - The extracts of leaf, root, stem, and the callus obtained from Pseudarthria viscida showed significant inhibitory activity against some fungal pathogens causing major diseases in crop plants and stored food grains. PMID- 15351114 TI - The antioxidant activity of some commonly used vegetables in Iranian diet. AB - Methanol extracts of 26 commonly used vegetables in Iranian diet were monitored for antioxidant activity against linoleic acid peroxidation. Some vegetable extracts including savory, radish leaf, garden-cress, spirmint, leek, chive (aerial part), lettuce and dill showed an antioxidant activity comparable with those of dl-alpha-tocopherol and quercetin. PMID- 15351115 TI - Antibacterial activity of Salacia beddomei. AB - Leaves and stems of Salacia beddomei were extracted successively with petroleum ether, ethyl acetate and chloroform and tested for their antibacterial activity. Ethyl acetate extracts were found to be most effective against all the tested organisms. PMID- 15351116 TI - Antioxidant compounds from Chaerophyllum hirsutum extracts. AB - Antioxidant activity of sequential extracts obtained from Chaerophyllum hirsutum was tested using the reaction with the stable radical 1,1 diphenyl-2 picrylhydrazyl (DPPH). Activity-guided fractionation led to the isolation of 6 methylluteolin, luteolin-7-O-beta-D-glucopyranoside, chlorogenic acid, ferulic acid, feruloyl-6-O-beta-D-glucopyranoside and maltol-beta-D-glucopyranoside. PMID- 15351117 TI - Anticariogenic activity of some tropical medicinal plants against Streptococcus mutans. AB - The methanol extracts of five tropical plants, Baeckea frutescens, Glycyrrhiza glabra, Kaempferia pandurata, Physalis angulata and Quercus infectoria, exhibited potent antibacterial activity against the cariogenic bacterium Streptococcus mutans. In particular, G. glabra, K. pandurata and P. angulata conferred fast killing bactericidal effect against S. mutans in 2 min at 50 microg/ml of extract concentration. PMID- 15351118 TI - Antibacterial activity of essential oils of Pimenta racemosa var. terebinthina and Pimenta racemosa var. grisea. AB - The antibacterial activity of essential oils of Pimenta racemosa var. terebinthina and P. racemosa var. grisea was determined against Gram (+) and Gram (-) bacteria. P. racemosa var. grisea demonstrated a more pronounced activity. These data would indicate the potential usefulness of the variety grisea as a microbiostatic, antiseptic or disinfectant agent. PMID- 15351119 TI - Antibacterial effect of Phellinus linteus against methicillin-resistant Staphylococcus aureus. AB - Methanol extract and its fractions (CHCl3, n-BuOH and H2O) of the fruit body of Phellinus linteus mushroom were investigated for antibacterial activity against methicillin-resistant Staphylococcus aureus. The n-BuOH fraction showed a good antibacterial activity (MIC, 63-125 microg/ml) against all tested strains. PMID- 15351120 TI - Constituents of the pericarp of Glyptopetalum sclerocarpum. AB - Two new compounds, 6-oxo-20-hydroxy-20-epi-tingenol (1) and 2R*,3R*,5S* trihydroxy-6R*-nonadecyltetrahydropyran-4-one (2), were isolated from the pericarp of Glyptopetalum sclerocarpum along with other 10 constituents. PMID- 15351121 TI - Cucurbita-5,23-diene-3beta,25-diol from Sicana odorifera. AB - The spectral data of a new triterpene, cucurbita-5,23-diene-3beta,25-diol, isolated from the seeds of Sicana odorifera, are reported. PMID- 15351122 TI - Lotaustralin from Rhodiola rosea roots. AB - Lotaustralin was isolated as a mixture of two diastereoisomeric forms from the methanol extract of Rhodiola rosea roots, together with the known compounds rosavin, rosarin, rosin, rosiridin, salidroside, and beta-sitosterol. The structure of lotaustralin was established by 1D and 2D-NMR spectroscopy, including 1H-1H COSY, NOESY, HMQC, and HMBC, and FAB and HR MS. PMID- 15351123 TI - Constituents of aromatic plants: elemicin. AB - No results of short-term or chronic toxicity studies have been found. Elemicin did induce UDS in hepatocytes from male rats. Studies on carcinogenicity were negative, but the 1'-hydroxy-metabolite of elemicin gave positive and negative results. The total intake of elemicin from essential oil seems to be limited. The main source of intake appears to be nutmeg. Further studies are needed to evaluate if the intake of elemicin may represent a health risk. PMID- 15351125 TI - Proceedings of the Fifth International Workshop on Information Processing in Cells and Tissues. September 8-11, 2003. Lausanne Switzerland. PMID- 15351126 TI - A spiking neuron model for synchronous flashing of fireflies. AB - Certain species of fireflies show a group behavior of synchronous flashing. Their synchronized and rhythmic flashing has received much attention among many researchers, and there has been a study of biological models for their entrainment of flashing. The synchronous behavior of fireflies resembles the firing synchrony of integrate-and-fire neurons with excitatory or inhibitory connections. This paper shows an analysis of spiking neurons specialized for a firefly flashing model, and provides simulation results of multiple neurons with various transmission delays and coupling strengths. It also explains flashing patterns of some firefly species and examines the synchrony conditions depending on transmission delays and coupling strengths. PMID- 15351127 TI - A role of burst firings in encoding of spatiotemporally-varying stimulus. AB - To investigate a role of burst firings of neurons in encoding of spatiotemporally varying stimulus, we focus on electrosensory system of a weakly electric fish. Weakly electric fish generates electric field around its body using electric organ discharge and can accurately detect the location of an object using the modulation of electric field induced by the object. We developed a model of fish body by which we numerically describe the spatiotemporal patterns of electric field around the fish body. We also made neural models of electroreceptor distributed on the fish body and of electrosensory lateral-line lobe (ELL) to investigate what kinds of information of electric field distorted by an object they detect. Here we show that the spatiotemporal features of electric field around the fish body are encoded by the timing of burst firings of ELL neurons. The information of object distance is extracted by the area of synchronous firings of neurons in a higher nucleus, torus semicircularis. PMID- 15351128 TI - A neural model of predictive recognition in form pathway of visual cortex. AB - We present a functional model of form pathway in visual cortex based on predictive coding scheme, in which the prediction is compared with feedforward signals filtered by two kinds of spatial resolution maps, broad and fine resolution map. We propose here the functional role of the prediction and of the two kinds of resolution maps in perception of object form in visual system. The prediction is represented based on memory of dynamical attractors in temporal cortex, categorized by an elemental figure in posterior temporal cortex. The prediction is generated by the feedforward signals of main neurons in broad resolution maps of V(1) and V(4), and then is compared with the feedforward signals of main neurons in fine resolution map of V(1) and V(4). PMID- 15351129 TI - Oscillatory network with self-organized dynamical connections for synchronization based image segmentation. AB - An oscillatory network of columnar architecture located in 3D spatial lattice was recently designed by the authors as oscillatory model of the brain visual cortex. Single network oscillator is a relaxational neural oscillator with internal dynamics tunable by visual image characteristics - local brightness and elementary bar orientation. It is able to demonstrate either activity state (stable undamped oscillations) or "silence" (quickly damped oscillations). Self organized nonlocal dynamical connections of oscillators depend on oscillator activity levels and orientations of cortical receptive fields. Network performance consists in transfer into a state of clusterized synchronization. At current stage grey-level image segmentation tasks are carried out by 2D oscillatory network, obtained as a limit version of the source model. Due to supplemented network coupling strength control the 2D reduced network provides synchronization-based image segmentation. New results on segmentation of brightness and texture images presented in the paper demonstrate accurate network performance and informative visualization of segmentation results, inherent in the model. PMID- 15351130 TI - A neural mechanism for detecting the distance of a selected target by modulating the FM sweep rate of biosonar in echolocation of bat. AB - Most species of bats making echolocation use frequency modulated (FM) ultrasonic pulses to measure the distance to targets. These bats detect with a high accuracy the arrival time differences between emitted pulses and their echoes generated by targets. In order to clarify the neural mechanism for echolocation, we present neural model of inferior colliculus (IC), medial geniculate body (MGB) and auditory cortex (AC) along which information of echo delay times is processed. The bats increase the downward frequency sweep rate of emitted FM pulse as they approach the target. The functional role of this modulation of sweep rate is not yet clear. In order to investigate the role, we calculated the response properties of our models of IC, MGB, and AC changing the target distance and the sweep rate. We found based on the simulations that the distance of a target in various ranges may be encoded the most clearly into the activity pattern of delay time map network in AC, when the sweep rate of FM pulse used is coincided with the observed value which the bats adopt for each range of target distance. PMID- 15351131 TI - Human-like reflex control for an artificial hand. AB - In this paper, we illustrate the low level reflex control used to govern an anthropomorphic artificial hand. The paper develops the position and stiffness control strategy based on dynamic artificial neurons able to simulate the neurons acting in the human reflex control. The controller has a hierarchical structure. At the lowest level there are the receptors able to convert the analogical signal into a neural impulsive signal appropriate to govern the reflex control neurons. Immediately upon it, the artificial motoneurons set the actuators inner pressure to control the finger joint position and moment. Other auxiliary neurons in combination with the motoneurons are able to set the finger stiffness and emulate the inverse myotatic reflex control. Stiffness modulation is important both to save energy during task execution, and to manage objects made of different materials. The inverse myotatic reflex is able to protect the hand from possible harmful external actions. The paper also presents the dynamic model of the joints and of the artificial muscles actuating Blackfingers, our artificial hand. This new type of neural control has been simulated on the Blackfingers model; the results indicate that the developed control is very flexible and efficient for all kind of joints present in the humanoid hand. PMID- 15351132 TI - Neuronal evolution: analysis of regulatory genes in a first-evolved nervous system, the hydra nervous system. AB - Cnidarians represent the first animal phylum with an organized nervous system and a complex active behavior. The hydra nervous system is formed of sensory motoneurons, ganglia neurons and mechanoreceptor cells named nematocytes, which all differentiate from a common stem cell. The neurons are organized as a nerve net and a subset of neurons participate in a more complex structure, the nerve ring that was identified in most cnidarian species at the base of the tentacles. In order to better understand the genetic control of this neuronal network, we analysed the expression of evolutionarily conserved regulatory genes in the hydra nervous system. The Prd-class homeogene prdl-b and the nuclear orphan receptor hyCOUP-TF are expressed at strong levels in proliferating nematoblasts, a lineage where they were found repressed during patterning and morphogenesis, and at low levels in distinct subsets of neurons. Interestingly, Prd-class homeobox and COUP TF genes are also expressed during neurogenesis in bilaterians, suggesting that mechanoreceptor and neuronal cells derive from a common ancestral cell. Moreover, the Prd-class homeobox gene prdl-a, the Antp-class homeobox gene msh, and the thrombospondin-related gene TSP1, which are expressed in distinct subset of neurons in the adult polyp, are also expressed during early budding and/or head regeneration. These data strengthen the fact that two distinct regulations, one for neurogenesis and another for patterning, already apply to these regulatory genes, a feature also identified in bilaterian related genes. PMID- 15351133 TI - The epitheliome: agent-based modelling of the social behaviour of cells. AB - We have developed a new computational modelling paradigm for predicting the emergent behaviour resulting from the interaction of cells in epithelial tissue. As proof-of-concept, an agent-based model, in which there is a one-to-one correspondence between biological cells and software agents, has been coupled to a simple physical model. Behaviour of the computational model is compared with the growth characteristics of epithelial cells in monolayer culture, using growth media with low and physiological calcium concentrations. Results show a qualitative fit between the growth characteristics produced by the simulation and the in vitro cell models. PMID- 15351134 TI - Coupled computational simulation and empirical research into the foraging system of Pharaoh's ant (Monomorium pharaonis). AB - The Pharaoh's ant (Monomorium pharaonis), a significant pest in many human environments, is phenomenally successful at locating and exploiting available food resources. Several pheromones are utilized in the self-organized foraging of this ant but most aspects of the overall system are poorly characterised. Agent based modelling of ants as individual complex X-machines facilitates study of the mechanisms underlying the emergence of trails and aids understanding of the process. Conducting simultaneous modelling, and simulation, alongside empirical biological studies is shown to drive the research by formulating hypotheses that must be tested before the model can be verified and extended. Integration of newly characterised behavioural processes into the overall model will enable testing of general theories giving insight into division of labour within insect societies. This study aims to establish a new paradigm in computational modelling applicable to all types of multi-agent biological systems, from tissues to animal societies, as a powerful tool to accelerate basic research. PMID- 15351135 TI - Mathematical model of retinal mosaic formation. AB - The retina is one of the best examples of modular organisation in neural circuitry. This modular structure enables it to perform parallel processing. A mathematical model of the retina has been set up, focusing on mechanical features of retinal neurons and on the interaction and dendritic overlapping among retinal cells. The model focuses on the actions of local mechanical forces on the neuron's cytoskeleton. The cytoskeleton is regarded as a structure in which elastic and rigid elements are combined according to the tensegrity concept. We have assumed that dendritic overlap takes place in such a way as to favour uniform retinal neurons' distribution and that dendritic overlap is the only cause of neuron's motion on the retinal surface. This overlap depends on the growth of the dendrites due to the cytoskeletic deformation. The results obtained are in agreement with experimental results that support the notion that local mechanical interaction and dendritic overlapping are capable to transform random cell distributions into regular mosaics. PMID- 15351136 TI - Parallelising a model of bacterial interaction and evolution. AB - Large simulations of bacterial colonies require huge amounts of computational time, the only way to achieve the necessary level of performance is with parallel computers and a suitably designed implementation that maps the problem onto the hardware. For real problems this mapping can be a non-trivial problem requiring careful consideration of the constraints in both the system being modelled and the hardware that executes that model. Here we describe an implementation of a system for modelling bacterial evolution that encompasses many physical scales. This system is composed entirely of individual entities all playing out a complex series of interactions. These individuals exist at the scale of the population of bacterial and at the gene product scale. This paper reports that it is possible to map a dynamic problem such as this onto fixed resources, for the most part making use of implicit multiplexing of resources provided by the OS and partitioning the problem to reduce communication time. Through this an efficient simulation can be created, making maximal use of the available hardware without constraining the model to require excessively specific resources. PMID- 15351137 TI - Spontaneous switching of frequency-locking by periodic stimulus in oscillators of plasmodium of the true slime mold. AB - Microfabrication technique was used to construct a model system with a living cell of plasmodium of the true slime mold, Physarum polycephalum, a living coupled oscillator system. Its parameters can be systematically controlled as in computer simulations, so that results are directly comparable to those of general mathematical models. As the first step, we investigated responses in oscillatory cells, the oscillators of the plasmodium, to periodic stimuli by temperature changes to elucidate characteristics of the cells as nonlinear systems whose internal dynamics are unknown because of their complexity. We observed that the forced oscillator of the plasmodium show 1:1, 2:1, 3:1 frequency locking inside so-called Arnold tongues regions as well as in other nonlinear systems such as chemical systems and other biological systems. In addition, we found spontaneous switching behavior from certain frequency locking states to other states, even under certain fixed parameters. This technique can be applied to more complex systems with multiple elements, such as coupled oscillator systems, and would be useful to investigate complicated phenomena in biological systems such as information processing. PMID- 15351138 TI - Immune system learning and memory quantified by graphical analysis of B lymphocyte phylogenetic trees. AB - The immune system learns from its encounters with pathogens and memorizes its experiences. One of the mechanisms it uses for this purpose is the intra individual evolution of antigen receptors on B lymphocytes, achieved via hypermutation and selection of antigen receptor variable region genes during an immune response. We have developed a novel method for analyzing the graphical properties of phylogenetic trees of receptor genes which have been mutated and selected during an immune response. In the study presented here, we address the artifacts introduced by experimental methods of cell collection for DNA analysis, the meaning of each parameter measured on the tree graphs, and the differences between the dynamics of the humoral immune response in different lymphoid tissues. PMID- 15351139 TI - Artificial cell division. AB - After a survey of the theory and some realizations of self-replicating machines, this paper presents a novel self-replicating loop endowed with universal construction and computation properties. Based on the hardware implementation of the so-called Tom Thumb algorithm, the design of this loop leads to a new kind of cellular automaton made of a processing and a control units. The self-replication of the Swiss flag serves as an artificial cell division example of the loop which, according to autopoietic evaluation criteria, corresponds to a cell showing the phenomenology of a living system. PMID- 15351140 TI - A computational model for cell differentiation. AB - In this paper, we present a word set generating mechanism, called cell differentiation system, inspired by the tissue process formation in multicellular organisms, which might model some properties of evolving communities of living cells at the syntactical level. The tools utilized to model these biological phenomena belong to the formal language theory. In this context chromosomal mutations are defined as operations on strings and the differentiation according to the control of gene expression is represented by some random-context conditions in formal languages. In the presented formal framework we prove that in a simplified form of this formalism, with only one cell-type which is regular, one single cell and no mitosis involved, the problem of establishing whether or not the set of vectors of integers indicating the number of cells in each population, is finite, linear or semilinear, is recursively undecidable. However, one can algorithmically decide whether or not a cell-differentiation system of finite cell-type can produce a specific generation of cells. PMID- 15351141 TI - An evolving ontogenetic cellular system for better adaptiveness. AB - In this paper, we present an original cellular system named Phuon. The main motivation behind this project is to go beyond classical cellular systems, such as cellular automata (CA). CA often lack adaptability and turn out to be very brittle in uncertain environment. The idea here is to add ontogeny to cellularity, growth and development being means of adaptation and thus robustness. However, we do not wish to develop yet another cellular system for the sake of it. What we are seeking in the long term is a developmental system for problem solving. This global aim enticed us into finding a way to map a desired global behavior of the system to the local behavior of a cell. Quite naturally a peculiar brand of genetic programming was used for that purpose. The results are still preliminary but in our view they already validate some of the hypotheses behind this work. PMID- 15351142 TI - POEtic: an electronic tissue for bio-inspired cellular applications. AB - In this paper, we introduce the general architecture of a new electronic tissue called POEtic. This reconfigurable circuit is designed to ease the implementation of bio-inspired systems that bring cellular applications into play. It contains special features that allow a developer to realize systems that require evolution (Phylogenesis), development (Ontogenesis), and/or learning (Epigenesis). A dynamic routing algorithm has been added to a structure similar to that of common commercial FPGAs, in order to allow the creation of data paths between cells. As the creation of these paths is dynamic, it is possible to add new cells or to repair faulty ones at runtime. PMID- 15351143 TI - Hardware optimization and serial implementation of a novel spiking neuron model for the POEtic tissue. AB - In this paper we describe the hardware implementation of a spiking neuron model, which uses a spike time dependent plasticity (STDP) rule that allows synaptic changes by discrete time steps. For this purpose an integrate-and-fire neuron is used with recurrent local connections. The connectivity of this model has been set to 24 neighbours, so there is a high degree of parallelism. After obtaining good results with the hardware implementation of the model, we proceed to simplify this hardware description, trying to keep the same behaviour. Some experiments using dynamic grading patterns have been used in order to test the learning capabilities of the model. Finally, the serial implementation has been realized. PMID- 15351144 TI - Coevolving solutions to the shortest common superstring problem. AB - The shortest common superstring (SCS) problem, known to be NP-Complete, seeks the shortest string that contains all strings from a given set. In this paper we compare four approaches for finding solutions to the SCS problem: a standard genetic algorithm, a novel cooperative-coevolutionary algorithm, a benchmark greedy algorithm, and a parallel coevolutionary-greedy approach. We show the coevolutionary approach produces the best results, and discuss directions for future research. PMID- 15351145 TI - Adapting genetic regulatory models by genetic programming. AB - In this paper, we focus on the task of adapting genetic regulatory models based on gene expression data from microarrays. Our approach aims at automatic revision of qualitative regulatory models to improve their fit to expression data. We describe a type of regulatory model designed for this purpose, a method for predicting the quality of such models, and a method for adapting the models by means of genetic programming. We also report experimental results highlighting the ability of the methods to infer models on a number of artificial data sets. In closing, we contrast our results with those of alternative methods, after which we give some suggestions for future work. PMID- 15351146 TI - Modelling biological evolvability: implicit context and variation filtering in enzyme genetic programming. AB - This paper describes recent insights into the role of implicit context within the representations of evolving artefacts and specifically within the program representation used by enzyme genetic programming. Implicit context occurs within self-organising systems where a component's connectivity is both determined implicitly by its own definition and is specified in terms of the behavioural context of other components. This paper argues that implicit context is an important source of evolvability and presents experimental evidence that supports this assertion. In particular, it introduces the notion of variation filtering, suggesting that the use of implicit context within representations leads to meaningful variation filtering whereby inappropriate change is ignored and meaningful change is encouraged during evolution. PMID- 15351147 TI - Towards more biological mutation operators in gene regulation studies. AB - Genetic regulation is often viewed as a complex system whose properties emerge from the interaction of regulatory genes. One major paradigm for studying the complex dynamics of gene regulation uses directed graphs to explore structure, behaviour and evolvability. Mutation operators used in such studies typically involve the insertion and deletion of nodes, and the insertion, deletion and rewiring of links at the network level. These network-level mutational operators are sufficient to allow the statistical analysis of network structure, but impose limitations on the way networks are evolved. There are a wide variety of mutations in DNA sequences that have yet to be analysed for their network-level effects. By modelling an artificial genome at the level of nucleotide sequences and mapping it to a regulatory network, biologically grounded mutation operators can be mapped to network-level mutations. This paper analyses five such sequence level mutations (single-point mutation, transposition, inversion, deletion and gene duplication) for their effects at the network level. Using analytic and simulation techniques, we show that it is rarely the case that nodes and links are cleanly added or deleted, with even the simplest point mutation causing a wide variety of network-level modifications. As expected, the vast majority of simple (single-point) mutations are neutral, resulting in a neutral plateau from which a range of functional behaviours can be reached. By analysing the effects of sequence-level mutations at the network level of gene regulation, we aim to stimulate more careful consideration of mutation operators in gene regulation models than has previously been given. PMID- 15351148 TI - Coding theory based models for protein translation initiation in prokaryotic organisms. AB - Our research explores the feasibility of using communication theory, error control (EC) coding theory specifically, for quantitatively modeling the protein translation initiation mechanism. The messenger RNA (mRNA) of Escherichia coli K 12 is modeled as a noisy (errored), encoded signal and the ribosome as a minimum Hamming distance decoder, where the 16S ribosomal RNA (rRNA) serves as a template for generating a set of valid codewords (the codebook). We tested the E. coli based coding models on 5' untranslated leader sequences of prokaryotic organisms of varying taxonomical relation to E. coli including: Salmonella typhimurium LT2, Bacillus subtilis, and Staphylococcus aureus Mu50. The model identified regions on the 5' untranslated leader where the minimum Hamming distance values of translated mRNA sub-sequences and non-translated genomic sequences differ the most. These regions correspond to the Shine-Dalgarno domain and the non-random domain. Applying the EC coding-based models to B. subtilis, and S. aureus Mu50 yielded results similar to those for E. coli K-12. Contrary to our expectations, the behavior of S. typhimurium LT2, the more taxonomically related to E. coli, resembled that of the non-translated sequence group. PMID- 15351149 TI - A theory of optimal differential gene expression. AB - We investigate a model of optimal regulation, intended to describe large-scale differential gene expression. Relations between the optimal expression patterns and the function of genes are deduced from an optimality principle: the regulators have to maximise a fitness function which they influence directly via a cost term, and indirectly via their control on important cell variables, such as metabolic fluxes. According to the model, the optimal linear response to small perturbations reflects the regulators' functions, namely their linear influences on the cell variables. The optimal behaviour can be realised by a linear feedback mechanism. Known or assumed properties of response coefficients lead to predictions about regulation patterns. A symmetry relation predicted for deletion experiments is verified with gene expression data. Where the optimality assumption is valid, our results justify the use of expression data for functional annotation and for pathway reconstruction and suggest the use of linear factor models for the analysis of gene expression data. PMID- 15351150 TI - An engineer's view on genetic information and biological evolution. AB - We develop ideas on genome replication introduced in Battail [Europhys. Lett. 40 (1997) 343]. Starting with the hypothesis that the genome replication process uses error-correcting means, and the auxiliary one that nested codes are used to this end, we first review the concepts of redundancy and error-correcting codes. Then we show that these hypotheses imply that: distinct species exist with a hierarchical taxonomy, there is a trend of evolution towards complexity, and evolution proceeds by discrete jumps. At least the first two features above may be considered as biological facts so, in the absence of direct evidence, they provide an indirect proof in favour of the hypothesized error-correction system. The very high redundancy of genomes makes it possible. In order to explain how it is implemented, we suggest that soft codes and replication decoding, to be briefly described, are plausible candidates. Experimentally proven properties of long-range correlation of the DNA message substantiate this claim. PMID- 15351151 TI - Evolving beyond perfection: an investigation of the effects of long-term evolution on fractal gene regulatory networks. AB - This paper continues a theme of exploring algorithms based on principles of biological development for tasks such as pattern generation, machine learning and robot control. Previous work has investigated the use of genes expressed as fractal proteins to enable greater evolvability of gene regulatory networks (GRNs). Here, the evolution of such GRNs is investigated further to determine whether evolution exhibits natural tendencies towards efficiency and graceful degradation of developmental programs. Experiments where "perfect" GRNs are evolved for a further thousand generations without the addition of any further selection pressure, confirm this hypothesis. After further evolution, the perfect GRNs operate in a more efficient manner (using fewer proteins) and show an improved ability to function correctly with missing genes. When the algorithm is applied to applications (e.g. robot control) this equates to efficient and fault tolerant controllers. PMID- 15351152 TI - Parallel computation of simple arithmetic using peptide-antibody interactions. AB - We propose a theoretical model for representing and manipulating binary numbers using peptide-antibody interactions. In particular, we present models to solve simple binary arithmetical problems like addition and subtraction. As the interactions can take place in parallel we show that the number of steps is independent of the size (bits) of the numbers. PMID- 15351153 TI - Is there a gender paradox in the early invasive strategy for non ST-segment elevation acute coronary syndromes? PMID- 15351154 TI - Glycoprotein IIb/IIIa inhibition and long-term benefit: the stuff of dreams? PMID- 15351155 TI - Heart failure disease management works, but will it succeed? PMID- 15351156 TI - Continental doctors take leadership in British medicine. PMID- 15351157 TI - The effectiveness of disease management programmes in reducing hospital re admission in older patients with heart failure: a systematic review and meta analysis of published reports. AB - AIMS: To systematically evaluate the published evidence regarding the effectiveness of disease management programmes (DMPs) reducing hospital re admissions among elderly patients with heart failure (HF). METHODS AND RESULTS: Computerised search of MEDLINE (1966 to 31 August 2003) and EMBASE (1966 to 31 August 2003). The Cochrane Library was also searched, and reference lists of review articles on the topic, and of all relevant studies identified, were scanned. Search and selection of studies, data-extraction using standardised forms, and assessment of study quality was performed by two reviewers. The end point was the proportion of persons who underwent hospital re-admission, and pooled relative risks (RR) were used to summarise the effectiveness of DMPs. The meta-analysis included 54 articles, comprising 27 randomised and 27 non randomised controlled studies. Randomised studies consistently suggested that, in comparison with usual care, DMP reduced the frequency of re-admission for HF or cardiovascular disease by 30% (pooled RR 0.70; confidence interval (CI) 95% 0.62 0.79), all-cause re-admission by 12% (pooled RR 0.88, 95% CI: 0.79-0.97), and the combined event of re-admission or death by 18% (pooled RR 0.82, 95% CI: 0.72 0.94). The results displayed no substantial variation when only DMPs with home visits, out-patient visits to a clinic, or patient follow-up longer than 6 months were included. For DMPs with out-patient clinical visits, however, the reduction in re-admission for HF or cardiovascular disease, and for all causes, did not attain statistical significance. The magnitude of DMP benefits reported by non randomised studies was more than double that reported by randomised studies. Practically all the non-randomised studies failed to control for confounding factors, such as severity, co-morbidity and drug therapy. CONCLUSION: DMPs are effective at reducing re-admissions among elderly patients with HF. Their effectiveness is close to that observed in clinical trials evaluating drugs for HF, such as angiotensin-converting enzyme inhibitors, beta-blockers or digoxin. However, since none of the DMP studies compared different interventions directly, we do not know the relative effectiveness of types of healthcare delivery within the DMP. PMID- 15351158 TI - Heart failure clinics and outpatient management: review of the evidence and call for quality assurance. AB - Despite major advances in treatment options for heart failure patients, morbidity and mortality remain unacceptably high. Frequent readmissions are distressful for patients and are associated with large costs for society. In an attempt to improve care for heart failure patients and thereby reduce morbidity and hospital readmissions, specialised heart failure clinics have emerged over the last 10 years. In particular, clinics relying, at least in part, on nurses specially trained in heart failure have gained popularity. This review of the published literature describes the wide variety of designs and the types of interventions taking place in such heart failure clinics. A total of 18 randomised studies comparing heart failure clinics using nurse intervention with conventional care have been published to date, and the majority of these have shown either a reduction in hospital readmissions or shortening of hospitalisations in the intervention group. These findings are supported by the results of several non randomised, controlled investigations. Thus, it is concluded that heart failure clinics using nurse intervention should be an integrated part of the care process for patients with heart failure wherever possible. We argue that ongoing attention should be paid to the quality of care delivered by the clinics to ensure that the benefit of this intervention strategy persists. Thus, it would be of importance to continuously record relevant data describing the care process using specific indicators such as ACE-inhibitor and beta-blocker use and doses. One possible, practical method to apply such continuous quality assurance may be by means of electronic medical record databases. PMID- 15351159 TI - Medical predictors for psychopathology in adults with operated congenital heart disease. AB - AIMS: To test the predictive value of medical variables, covering the complete medical course from birth until the present, for long-term behavioural and emotional problems in adulthood, in patients operated for congenital heart disease in childhood. METHODS AND RESULTS: This study concerns the second psychological and medical follow-up of a cohort of patients operated for congenital heart disease (n=362; age 20-46 years). Behavioural and emotional problems were assessed with the Young Adult Self-Report and the Young Adult Behavior Checklist. Medical prediction variables were derived from medical examination and file search. Being female, having low exercise capacity and restrictions imposed by physicians are significant predictors for behavioural and emotional problems as reported by patients themselves. Regarding the scar, personal experiences of patients form a better predictor for later problems than judgement of aesthetical aspects by physicians. Early hospitalisations with reoperations are predictive for behavioural and emotional problems as reported by other informants. The cardiac diagnoses of ventricular septal defect and transposition of the great arteries are associated with higher levels of behavioural and emotional problems. CONCLUSION: Recent experiences concerning the scar, physical condition and imposed restrictions are the strongest predictors for behavioural and emotional problems as reported by patients themselves. PMID- 15351160 TI - Quantifying the heart failure epidemic: prevalence, incidence rate, lifetime risk and prognosis of heart failure The Rotterdam Study. AB - AIMS: To determine the prevalence, incidence rate, lifetime risk and prognosis of heart failure. METHODS AND RESULTS: The Rotterdam Study is a prospective population-based cohort study in 7983 participants aged > or =55. Heart failure was defined according to criteria of the European Society of Cardiology. Prevalence was higher in men and increased with age from 0.9% in subjects aged 55 64 to 17.4% in those aged > or =85. Incidence rate of heart failure was 14.4/1000 person-years (95% CI 13.4-15.5) and was higher in men (17.6/1000 man-years, 95% CI 15.8-19.5) than in women (12.5/1000 woman-years, 95% CI 11.3-13.8). Incidence rate increased with age from 1.4/1000 person-years in those aged 55-59 to 47.4/1000 person-years in those aged > or =90. Lifetime risk was 33% for men and 29% for women at the age of 55. Survival after incident heart failure was 86% at 30 days, 63% at 1 year, 51% at 2 years and 35% at 5 years of follow-up. CONCLUSION: Prevalence and incidence rates of heart failure are high. In individuals aged 55, almost 1 in 3 will develop heart failure during their remaining lifespan. Heart failure continues to be a fatal disease, with only 35% surviving 5 years after the first diagnosis. PMID- 15351161 TI - The pro-apoptotic serum activity is an independent mortality predictor of patients with heart failure. AB - AIM: Systemic inflammation with elevated serum levels of circulating pro inflammatory cytokines is a major determinant of prognosis in heart failure (HF). Since serum of patients with HF induces apoptosis of endothelial cells (EC), we aimed to determine whether the pro-apoptotic activity in the serum may predict prognosis of patients with HF. METHODS AND RESULTS: We measured the pro-apoptotic activity in the serum of 48 patients with HF of different aetiology by an ex vivo cell culture assay and subsequently monitored these patients for the single endpoint all-cause mortality. During follow-up, 16 patients died and 11 patients received a heart transplant. Survivors had a lower pro-apoptotic serum activity (P=0.007). By univariate analysis, pro-apoptotic serum activity, NYHA class, pro BNP, low blood pressure, and creatinine levels were significantly associated with mortality. In a multivariable stepwise Cox-regression model, the pro-apoptotic serum activity (adjusted hazard ratio, HR=1.85 per %, P=0.008), elevated pro-BNP levels (HR=9.35 per log[pro-BNP], P=0.001), and low blood pressure (HR=0.96 per mmHg, P=0.041) remained as independent predictors of death. CONCLUSION: In this exploratory study, the pro-apoptotic serum capacity is independently associated with a worse prognosis in patients with HF, suggesting that the assessment of serum-induced EC apoptosis could provide an integrative estimate of the deleterious effects of various pro-inflammatory cytokines and other cytotoxic factors in HF. PMID- 15351162 TI - Reduction of cyclosporine after introduction of mycophenolate mofetil improves chronic renal dysfunction in heart transplant recipients--the IMPROVED multi centre study. AB - AIMS: This comparative prospective multi-centre study evaluated efficacy and safety of cyclosporine A downtitration in heart transplant recipients with chronic renal dysfunction potentially attributable to cyclosporine (n=161). METHODS: In the intervention arm (n=109, recruited from 9 centres), mycophenolate mofetil was introduced de novo or substituting azathioprine, followed by cyclosporine reduction (target trough levels 2-4 microg/ml and 50 ng/ml, respectively). In controls (n=52, recruited from 1 centre), immunosuppression remained unchanged. Renal function was recorded twelve, six, and three months before, and throughout the eight-month study period. RESULTS: At study entry, cyclosporine trough levels and renal function parameters were comparable. At study end, mean+/-SD cyclosporine in the intervention arm was 57+/-24 vs. 116+/ 36 ng/ml in controls. During the study, creatinine decreased by 23.3+/-50.7 micromol/l (P<0.0001) in the intervention arm but increased by 7.3+/-46.9 micromol/l (P=0.992) in controls (P=0.0001 for comparison between groups). A creatinine reduction of at least 20% was found in 35% of subjects of the intervention arm but only in 4% in the control arm (P<0.0001 for comparison between groups). Improvement in renal function was not weakened after adjustment for baseline characteristics in multiple regression analysis. Renal function improved in strata of creatinine entry values from 150 to 310 micromol/l, regardless of the presence of diabetes. Myocardial biopsies at target levels for cyclosporine and mycophenolate mofetil showed three reversible subclinical rejection episodes. CONCLUSIONS: Cyclosporine downtitration improved renal dysfunction in diabetic and non-diabetic heart transplant recipients across a wide range of creatinine levels. The long-term benefit of this strategy deserves further study. PMID- 15351163 TI - Five-year outcome of patients with acute myocardial infarction enrolled in a randomised trial assessing the value of abciximab during coronary artery stenting. AB - AIMS: The aim of the study was to investigate the long-term (five years) efficacy of glycoprotein IIb/IIIa inhibition with abciximab given as an adjunct therapy to coronary stenting in patients with acute myocardial infarction (MI) using the patient cohort of the Intracoronary Stenting and Antithrombotic Regimen-2 (ISAR 2) randomised trial. METHODS AND RESULTS: The patient cohort of ISAR-2 trial (401 patients) was followed up for 5 years after enrollment. There were 201 patients in the abciximab group (stenting plus abciximab) and 200 patients in the control group (stenting without abciximab). The primary end-point of the study was mortality at 5 years. Recurrent MI and target vessel re-vascularisation were also assessed at 5 years after enrollment. On the basis of the Kaplan-Meier analyses, the 5-year mortality was 17.8% (35 patients) in the group with abciximab and 14.6% (29 patients) in the control group (relative risk, 1.20 [95% confidence interval, 0.73-1.96]; P=0.47). The 5-year combined incidence of death, recurrent MI and target vessel re-vascularisation was 38.2% (76 patients) in the group of abciximab and 37.7% (75 patients) in the control group (relative risk, 0.97 [95% confidence interval, 0.70-1.33]; P=0.83). Multivariable analysis showed no significant independent association of abciximab with 5-year mortality (adjusted hazard ratio, 1.16 [95% confidence interval, 0.70-1.92]; P=0.55). CONCLUSION: These findings are not in support of a sustained clinical benefit at 5 years with the use of abciximab during coronary artery stenting in patients with acute MI. PMID- 15351164 TI - Do men benefit more than women from an interventional strategy in patients with unstable angina or non-ST-elevation myocardial infarction? The impact of gender in the RITA 3 trial. AB - AIMS: The RITA 3 trial randomized patients with non-ST-elevation myocardial infarction or unstable angina to strategies of early intervention (angiography followed by revascularization) or conservative care (ischaemia or symptom driven angiography). The aim of this analysis was to investigate the impact of gender on the effect of these two strategies. METHODS AND RESULTS: In total, 1810 patients (682 women and 1128 men) were randomized. The risk factor profile of women at presentation was markedly different to men. There was evidence that men benefited more from an early intervention strategy for death or non-fatal myocardial infarction at 1 year (adjusted odds ratios 0.63, 95% confidence interval 0.41 0.98 for men and 1.79, 95% confidence interval 0.95-3.35 for women; interaction p value=0.007). Men who underwent the assigned angiogram were more likely to be put forward for coronary artery bypass surgery, even after allowing for differences in disease severity. CONCLUSION: An early intervention strategy resulted in a beneficial effect in men which was not seen in women although caution is needed in interpretation. Further research is needed to evaluate why women do not appear to benefit from early intervention and to identify treatments that improve the prognosis of women. PMID- 15351165 TI - Responsiveness to inhaled nitric oxide is a predictor for mid-term survival in adult patients with congenital heart defects and pulmonary arterial hypertension. AB - BACKGROUND: It was previously demonstrated that in adults with congenital heart disease and pulmonary arterial hypertension and/or the Eisenmenger syndrome, the pulmonary circulation remained responsive to inhaled nitric oxide (iNO). We wanted to evaluate whether the responsiveness to iNO was related to mid-term outcome in these patients. METHODS: In 21 consecutive patients, total pulmonary vascular resistance (TPR) was measured at baseline, after 5 min iNO (80 ppm), and after NO withdrawal. Patients were considered responders when TPR was reduced by at least 20% during NO inhalation or when TPR increased by more than 10% after NO withdrawal. Responders and non-responders were followed prospectively and the primary endpoint of the study was cardiopulmonary death, the secondary endpoint the combination of death, need for treatment with prostacyclin or heart-lung transplantation. Kaplan-Meier survival curves for both groups were plotted and compared using log rank testing. RESULTS: Ten patients were considered responders (four male, median age 25 years, Q1 19 and Q3 66 years), while 11 patients did not respond (two male, median age 27 years, Q1 18 and Q3 40 years). The median follow-up time of the total group was 5.0 years (Q1 3.2 and Q3 5.7 years). Four of the non-responders died a cardiovascular death; none of the responders died. The difference in survival between responders and non-responders was statistically significant. For the secondary endpoint, no significant differences were found between both groups. CONCLUSIONS: The responsiveness to inhaled NO in adult patients with pulmonary arterial hypertension and/or the Eisenmenger syndrome is related to mid-term outcome. These findings might be important for risk stratification and the choice of treatment in this specific patient population. PMID- 15351166 TI - Detection of coronary artery disease by magnetic resonance myocardial perfusion imaging with various contrast medium doses: first European multi-centre experience. AB - AIMS: Magnetic resonance (MR) first-pass myocardial perfusion imaging during hyperaemia detects coronary artery stenoses in humans with test sensitivity depending on contrast medium (CM)-induced signal change in myocardium. In this prospective multi-centre study, the effect of CM dose on myocardial signal change and on diagnostic performance was evaluated using a stress-only approach. METHODS AND RESULTS: Ninety-four patients with known or suspected coronary artery disease (CAD) were randomised to 0.05,0.10, or 0.15 mmol/kg body weight of an extravascular CM (Gd-DTPA) and X-ray coronary angiography was performed within 30 days prior/after the MR examination. A multi-slice MR technique with identical hardware and software in all centres was used during hyperaemia (adenosine 0.14 mg/kg/min) to monitor myocardial CM wash-in kinetics and data were analysed semi automatically in a core laboratory. Protocol violations resulted in 80 complete studies with CAD (defined as > or =1 vessel with diameter stenosis > or =50% on quantitative coronary angiography) present in 19/29, 13/24, and 20/27 patients for doses 1, 2, and 3, respectively. In normal myocardium, the upslope increased with CM dose (overall-p<0.0001, ANOVA). For CAD detection the area under the receiver operator characteristics curve for subendocardial data (3 slices with quality score<4 representing 86% of cases) was 0.91+/-0.07 and 0.86+/-0.08 for doses 2 and 3, respectively, and was lower for dose 1 (0.53+/-0.13, p<0.01 and p<0.02 vs. doses 2 and 3, respectively). Corresponding sensitivities/specificities (95% confidence intervals) for pooled doses 2/3 were 93% (77-99%; ns vs. dose 1) and 75% (48-92%;p<0.05 vs. dose 1), respectively. CONCLUSIONS: With increasing doses of CM, a higher signal response in the myocardium was achieved and consequently this stress-only protocol, with CM doses of 0.10-0.15 mmol/kg combined with a semi-automatic analysis, yielded a high diagnostic performance for the detection of CAD. PMID- 15351167 TI - Animal research is a source of human compassion, not shame. PMID- 15351168 TI - Clean water alone cannot prevent disease. PMID- 15351169 TI - Contemplating ACTION--long-acting nifedipine in stable angina. PMID- 15351170 TI - Endovascular aneurysm repair. PMID- 15351171 TI - Lymph-node-negative oestrogen-receptor-positive breast cancer. PMID- 15351172 TI - Chandipura virus: an emerging human pathogen? PMID- 15351173 TI - Food colourings and preservatives--allergy and hyperactivity. PMID- 15351174 TI - Cancer and psychosocial distress: frequent companions. PMID- 15351175 TI - New insight into sudden infant-death syndrome. PMID- 15351176 TI - The trials of anakinra. PMID- 15351177 TI - Historical keywords: circulation. PMID- 15351179 TI - Women in medicine. PMID- 15351180 TI - Women in medicine. PMID- 15351181 TI - Medical historians and the tobacco industry. PMID- 15351183 TI - Fighting against tobacco companies. PMID- 15351184 TI - Fighting against tobacco companies. PMID- 15351185 TI - Medical historians and the tobacco industry. PMID- 15351186 TI - B-Aware: recall of intraoperative events. PMID- 15351187 TI - B-Aware: recall of intraoperative events. PMID- 15351189 TI - Analgesia in prostate biopsy. PMID- 15351190 TI - Marijuana on the state. PMID- 15351191 TI - Comparison of endovascular aneurysm repair with open repair in patients with abdominal aortic aneurysm (EVAR trial 1), 30-day operative mortality results: randomised controlled trial. AB - BACKGROUND: Endovascular aneurysm repair (EVAR) is a new technology to treat patients with abdominal aortic aneurysm (AAA) when the anatomy is suitable. Uncertainty exists about how endovascular repair compares with conventional open surgery. EVAR trial 1 was instigated to compare these treatments in patients judged fit for open AAA repair. METHODS: Between 1999 and 2003, 1082 elective (non-emergency) patients were randomised to receive either EVAR (n=543) or open AAA repair (n=539). Patients aged at least 60 years with aneurysms of diameter 5.5 cm or more, who were fit enough for open surgical repair (anaesthetically and medically well enough for the procedure), were recruited for the study at 41 British hospitals proficient in the EVAR technique. The primary outcome measure is all-cause mortality and these results will be released in 2005. The primary analysis presented here is operative mortality by intention to treat and a secondary analysis was done in per-protocol patients. FINDINGS: Patients (983 men, 99 women) had a mean age of 74 years (SD 6) and mean AAA diameter of 6.5 cm (SD 1). 1047 (97%) patients underwent AAA repair and 1008 (93%) received their allocated treatment. 30-day mortality in the EVAR group was 1.7% (9/531) versus 4.7% (24/516) in the open repair group (odds ratio 0.35 [95% CI 0.16-0.77], p=0.009). By per-protocol analysis, 30-day mortality for EVAR was 1.6% (8/512) versus 4.6% (23/496) for open repair (0.33 [0.15-0.74], p=0.007). Secondary interventions were more common in patients allocated EVAR (9.8% vs 5.8%, p=0.02). INTERPRETATION: In patients with large AAAs, treatment by EVAR reduced the 30-day operative mortality by two-thirds compared with open repair. Any change in clinical practice should await durability and longer term results. PMID- 15351192 TI - Effect of long-acting nifedipine on mortality and cardiovascular morbidity in patients with stable angina requiring treatment (ACTION trial): randomised controlled trial. AB - BACKGROUND: Calcium antagonists are widely prescribed for angina pectoris but their effect on clinical outcome is controversial. We aimed to investigate the effect of the calcium antagonist nifedipine on long-term outcome in patients with stable angina pectoris. METHODS: We randomly assigned 3825 patients with treated stable symptomatic coronary disease to double-blind addition of nifedipine GITS (gastrointestinal therapeutic system) 60 mg once daily and 3840 to placebo. The primary endpoint was the combination of death, acute myocardial infarction, refractory angina, new overt heart failure, debilitating stroke, and peripheral revascularisation. Mean follow-up was 4.9 years (SD 1.1). Analysis was by intention to treat. FINDINGS: 310 patients allocated nifedipine died (1.64 per 100 patient-years) compared with 291 people allocated placebo (1.53 per 100 patient-years; hazard ratio 1.07 [95% CI 0.91-1.25], p=0.41). Primary endpoint rates were 4.60 per 100 patient-years for nifedipine and 4.75 per 100 patient years for placebo (0.97 [0.88-1.07], p=0.54). With nifedipine, rate of death and any cardiovascular event or procedure was 9.32 per 100 patient-years versus 10.50 per 100 patient-years for placebo (0.89 [0.83-0.95], p=0.0012). The difference was mainly attributable to a reduction in the need for coronary angiography and interventions in patients assigned nifedipine, despite an increase in peripheral revascularisation. Nifedipine had no effect on the rate of myocardial infarction. INTERPRETATION: Addition of nifedipine GITS to conventional treatment of angina pectoris has no effect on major cardiovascular event-free survival. Nifedipine GITS is safe and reduces the need for coronary angiography and interventions. PMID- 15351193 TI - Treatment of lymph-node-negative, oestrogen-receptor-positive breast cancer: long term findings from National Surgical Adjuvant Breast and Bowel Project randomised clinical trials. AB - BACKGROUND: Findings from the National Surgical Adjuvant Breast and Bowel Project B-14 and B-20 trials showed that tamoxifen benefited women with oestrogen receptor-positive tumours and negative axillary nodes, and that chemotherapy plus tamoxifen was more effective than tamoxifen alone. We present long-term findings from those trials and relate them to age, menopausal status, and tumour oestrogen receptor concentrations. We also discuss the extent of progress made in the treatment of such patients. METHODS: B-14 patients were randomly assigned to placebo (n=1453) or tamoxifen (n=1439); B-20 patients to tamoxifen (n=788) or cyclophosphamide, methotrexate, fluorouracil, and tamoxifen (CMFT, n=789). Primary endpoints were recurrence-free survival and overall survival estimated according to patients' age, menopausal status, and tumour oestrogen-receptor concentration. Smoothed recurrence rates were used to measure patterns of recurrence as a continuous function of age. FINDINGS: Compared with placebo, tamoxifen benefited women in B-14 through 15 years, irrespective of age, menopausal status, or tumour oestrogen-receptor concentration (hazard ratio [HR] for recurrence-free survival 0.58, 95% CI 0.50-0.67, p<0.0001; HR for overall survival 0.80, 0.71-0.91, p=0.0008). In B-20, the benefit from CMFT over 12 years was greater than that from tamoxifen alone (HR for recurrence-free survival 0.52, 0.39-0.68, p<0.0001; HR for overall survival 0.78, 0.60-1.01, p=0.063). When CMFT was compared with placebo, there were reductions in treatment failure of about 65% in all age-groups. INTERPRETATION: Much benefit has been achieved in treatment of women with oestrogen-receptor-positive tumours and negative nodes. When planning systemic therapy for such patients of all ages, it should be understood that some have tumours with variable concentrations of oestrogen receptors, a surrogate for other biomarkers associated with tumour growth and response to treatment. Older women tend to have higher tumour oestrogen-receptor concentrations and are more likely to benefit from tamoxifen than from chemotherapy; in younger women, the converse is true. Consequently, the notion that use of tamoxifen or chemotherapy should be based only on age is too restrictive. PMID- 15351194 TI - A large outbreak of acute encephalitis with high fatality rate in children in Andhra Pradesh, India, in 2003, associated with Chandipura virus. AB - BACKGROUND: An outbreak of acute encephalitis of unknown origin with high case fatality (183 of 329 cases) was reported in children from Andhra Pradesh state in southern India during 2003. We investigated the causative agent. METHODS: Cell lines and peripheral blood lymphocyte co-cultures were used to isolate the causative agent from clinical samples. Identity of the agent was established by electron microscopy and serological and molecular assays. FINDINGS: Clinical samples tested negative for IgM antibodies to Japanese encephalitis, West Nile, dengue, and measles viruses, and for RNA of coronavirus, paramyxovirus, enterovirus, and influenza viruses. Virus was isolated from six patients with encephalitis and was identified as Chandipura virus by electron microscopy, complement fixation, and neutralisation tests. Chandipura virus RNA was detected in clinical samples from nine patients. Sequencing of five of these RNA samples showed 96.7-97.5% identity with the reference strain of 1965. Chandipura viral antigen and RNA were detected in brain tissue of a deceased child by immunofluorescent antibody test and PCR. Neutralising, IgG, and IgM antibodies to Chandipura virus were present in some patients' serum samples. Serum samples obtained after 4 days of illness were more frequently positive for IgM to Chandipura virus than were those obtained earlier (p<0.001). A similar trend was noted for neutralising antibodies. INTERPRETATION: Our findings suggest that this outbreak of acute encephalitis in Andhra Pradesh was associated with Chandipura virus, adding to the evidence suggesting that this virus should be considered as an important emerging pathogen. PMID- 15351195 TI - Parkinsonism, premature menopause, and mitochondrial DNA polymerase gamma mutations: clinical and molecular genetic study. AB - BACKGROUND: Mutations in the gene encoding mitochondrial DNA polymerase gamma (POLG), the enzyme that synthesises mitochondrial DNA (mtDNA), have been associated with a mitochondrial disease-autosomal dominant or recessive progressive external ophthalmoplegia-and multiple deletions of mtDNA. Mitochondrial dysfunction is also suspected to participate in the pathogenesis of Parkinson's disease. However, no primary gene defects affecting mitochondrial proteins causing mendelian transmission of parkinsonism have been characterised. We aimed to analyse the gene sequence of POLG in patients with progressive external ophthalmoplegia and their healthy relatives. METHODS: In seven families of various ethnic origins we assessed patients with progressive external ophthalmoplegia and unaffected individuals by clinical, biochemical, morphological, and molecular genetic characterisation and positron emission tomography (PET). FINDINGS: We recorded mutations in POLG in members of all seven families. Clinical assessment showed significant cosegregation of parkinsonism with POLG mutations (p<0.0001), and PET findings were consistent with dopaminergic neuron loss. Post-mortem examination in two individuals showed loss of pigmented neurons and pigment phagocytosis in substantia nigra without Lewy bodies. Furthermore, most women with progressive external ophthalmoplegia had early menopause-before age 35 years. The POLG gene defect resulted in secondary accumulation of mtDNA deletions in patients' tissues. INTERPRETATION: Dysfunction of mitochondrial POLG causes a severe progressive multisystem disorder including parkinsonism and premature menopause, which are not typical of mitochondrial disease. Cosegregation of parkinsonism and POLG mutations in our families suggests that when defective, this gene can underlie mendelian transmission of parkinsonism. RELEVANCE TO PRACTICE: Awareness that mitochondrial POLG mutations can underlie parkinsonism is important for clinicians working in diagnosis of movement disorders, as well as for studies of the genetics of Parkinson's disease. Further, progressive external ophthalmoplegia with muscle weakness and neuropathy can mask symptoms of parkinsonism, and clinicians should pay special attention to detect and treat parkinsonism in those individuals. PMID- 15351196 TI - Management of severe COPD. AB - Chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and mortality worldwide, and the burden of the disorder will continue to increase over the next 20 years despite medical intervention. Apart from smoking cessation, no approach or agent affects the rate of decline in lung function and progression of the disease. Especially in the later phase, COPD is a multicomponent disorder, and various integrated intervention strategies are needed as part of the optimum management programme. This seminar describes largely non-pharmacological interventions aimed at improving health status and function of disabled patients. Exacerbations become progressively more troublesome as baseline lung function declines, commonly necessitating hospital admission and associated with the development of acute respiratory failure. PMID- 15351197 TI - COPD is not a death sentence. PMID- 15351198 TI - Atkins and other low-carbohydrate diets: hoax or an effective tool for weight loss? AB - CONTEXT: The Atkins diet books have sold more than 45 million copies over 40 years, and in the obesity epidemic this diet and accompanying Atkins food products are popular. The diet claims to be effective at producing weight loss despite ad-libitum consumption of fatty meat, butter, and other high-fat dairy products, restricting only the intake of carbohydrates to under 30 g a day. Low carbohydrate diets have been regarded as fad diets, but recent research questions this view. STARTING POINT: A systematic review of low-carbohydrate diets found that the weight loss achieved is associated with the duration of the diet and restriction of energy intake, but not with restriction of carbohydrates. Two groups have reported longer-term randomised studies that compared instruction in the low-carbohydrate diet with a low-fat calorie-reduced diet in obese patients (N Engl J Med 2003; 348: 2082-90; Ann Intern Med 2004; 140: 778-85). Both trials showed better weight loss on the low-carbohydrate diet after 6 months, but no difference after 12 months. WHERE NEXT?: The apparent paradox that ad-libitum intake of high-fat foods produces weight loss might be due to severe restriction of carbohydrate depleting glycogen stores, leading to excretion of bound water, the ketogenic nature of the diet being appetite suppressing, the high protein content being highly satiating and reducing spontaneous food intake, or limited food choices leading to decreased energy intake. Long-term studies are needed to measure changes in nutritional status and body composition during the low carbohydrate diet, and to assess fasting and postprandial cardiovascular risk factors and adverse effects. Without that information, low-carbohydrate diets cannot be recommended. PMID- 15351199 TI - Overcoming health-systems constraints to achieve the Millennium Development Goals. AB - Effective interventions exist for many priority health problems in low income countries; prices are falling, and funds are increasing. However, progress towards agreed health goals remains slow. There is increasing consensus that stronger health systems are key to achieving improved health outcomes. There is much less agreement on quite how to strengthen them. Part of the challenge is to get existing and emerging knowledge about more (and less) effective strategies into practice. The evidence base also remains remarkably weak, partly because health-systems research has an image problem. The forthcoming Ministerial Summit on Health Research seeks to help define a learning agenda for health systems, so that by 2015, substantial progress will have been made to reducing the system constraints to achieving the MDGs. PMID- 15351200 TI - Two errors. PMID- 15351201 TI - Recurrent hyperkalaemia cured by amputation. PMID- 15351202 TI - Different ways to reach the top of a cell. Analysis of rotavirus assembly and targeting in human intestinal cells reveals an original raft-dependent, Golgi independent apical targeting pathway. PMID- 15351203 TI - Detection and characterization of betaretroviral sequences, related to sheep Jaagsiekte virus, in Africans from Nigeria and Cameroon. AB - Jaagsiekte retrovirus (JSRV) causes ovine pulmonary adenomatosis (OPA) that resembles bronchioloalveolar carcinoma (BAC) in humans. To test the possible role of JSRV in human diseases, DNA specimens from 103 individuals either healthy or suffering from lung carcinomas were analyzed for JSRV sequences. orf-x sequences were detected in 19 of 64 samples and gag-prt sequences in 4 of 38 samples, predominantly in individuals from Africa. Sequences obtained from orf-x amplimers varied in-between each other and differed from control endogenous ovine JSRV sequence. No association with lung cancer was found. This is the first report of JSRV-like sequences detected in humans. PMID- 15351204 TI - Importance of Akt signaling pathway for apoptosis in SARS-CoV-infected Vero E6 cells. AB - Severe acute respiratory syndrome (SARS) is an acute respiratory tract infectious disease that is associated with a new coronavirus (SARS-CoV). Our recent study indicated that SARS-CoV infection induces activation of the p38 mitogen-activated protein kinase (MAPK) signaling pathway and the p38 MAPK inhibitor partially inhibited its cytopathic effect in Vero E6 cells. The results of the present study indicated that before cell death, Akt, which is an inhibitor of apoptosis, was also activated in response to viral replication. Phosphorylation of a serine residue on Akt was detected at least 8 h postinfection (hpi), which declined after 18 hpi. Thus, the phosphatidylinositol 3-kinase (PI3K)/Akt pathway is activated in virus-infected Vero E6 cells. However, a threonine residue was not phosphorylated. A downstream target of Akt, glycogen synthase kinase 3beta (GSK 3beta), was slightly phosphorylated, indicating that the level of activation of Akt was very low. PKCzeta, which is downstream of the PI3K pathway, was also phosphorylated in virus-infected cells. These results suggested that weak activation of Akt cannot prevent apoptosis induced by SARS-CoV infection in Vero E6 cells. PMID- 15351205 TI - Analysis of the interaction between respiratory syncytial virus and lipid-rafts in Hep2 cells during infection. AB - The assembly of respiratory syncytial virus (RSV) in lipid-rafts was examined in Hep2 cells. Confocal and electron microscopy showed that during RSV assembly, the cellular distribution of the complement regulatory proteins, decay accelerating factor (CD55) and CD59, changes and high levels of these cellular proteins are incorporated into mature virus filaments. The detergent-solubility properties of CD55, CD59, and the RSV fusion (F) protein were found to be consistent with each protein being located predominantly within lipid-raft structures. The levels of these proteins in cell-released virus were examined by immunoelectronmicroscopy and found to account for between 5% and 15% of the virus attachment (G) glycoprotein levels. Collectively, our findings suggest that an intimate association exists between RSV and lipid-raft membranes and that significant levels of these host-derived raft proteins, such as those regulating complement activation, are subsequently incorporated into the envelope of mature virus particles. PMID- 15351206 TI - Adenosine receptors control HIV-1 Tat-induced inflammatory responses through protein phosphatase. AB - Recently, adenosine has been proposed to be a "metabolic" switch that may sense and direct immune and inflammatory responses. Inflammation and pro-inflammatory cytokine production are important in development of HIV-1 associated dementia, a devastating consequence of HIV-1 infection of the CNS. The HIV-1 protein Tat induces cell death in the CNS and activates local inflammatory responses partially by inducing calcium release from the endoplasmic reticulum. Because activation of adenosine receptors decreases production of the pro-inflammatory cytokine TNF-alpha in several experimental paradigms both in vitro and in vivo, we hypothesized that adenosine receptor activation would control both increased intracellular calcium and TNF-alpha production induced by Tat. Treatment of primary monocytes with Tat significantly increased the levels of intracellular calcium released from IP3 stores. Activation of adenosine receptors with CGS 21680 inhibited Tat-induced increases of intracellular calcium by 90 +/- 8% and was dependent on protein phosphatase activity because okadaic acid blocked the actions of CGS 21680. Tat-induced TNF-alpha production was inhibited 90 +/- 6% by CGS 21680 and concurrent treatment with okadaic acid blocked the inhibitory actions of CGS 21680. Using a model monocytic cell line, CGS 21680 treatment increased cytosolic serine/threonine phosphatase. Together, these data indicate that A2A receptor activation increases protein phosphatase activity, which blocks IP3 receptor-regulated calcium release and reduction of intracellular calcium inhibits TNF-alpha production in monocytes. PMID- 15351207 TI - Subgenomic RNA as a riboregulator: negative regulation of RNA replication by Barley yellow dwarf virus subgenomic RNA 2. AB - Barley yellow dwarf virus (BYDV) generates three 3'-coterminal subgenomic RNAs (sgRNAs) in infected cells. Translation of BYDV genomic RNA (gRNA) and sgRNA1 is mediated by the BYDV cap-independent translation element (BTE) in the 3' untranslated region. sgRNAs 2 and 3 are unlikely to be mRNAs. We proposed that accumulation of sgRNA2, which contains the BTE in its 5' UTR, regulates BYDV replication by trans-inhibiting translation of the viral polymerase from genomic RNA (gRNA). Here, we tested this hypothesis and found that: (i) co-inoculation of the BTE or sgRNA2 with BYDV RNA inhibits BYDV RNA accumulation in protoplasts; (ii) Brome mosaic virus (BMV), engineered to contain the BTE, trans-inhibits BYDV replication; and (iii) sgRNA2 generated during BYDV infection trans-inhibits both GFP expression from BMV RNA and translation of a non-viral reporter mRNA. We conclude that sgRNA2, via its BTE, functions as a riboregulator to inhibit translation of gRNA. This may make gRNA available as a replicase template and for encapsidation. Thus, BYDV sgRNA2 joins a growing list of trans-acting regulatory RNAs. PMID- 15351208 TI - Characterization of a nuclear localization signal in the C-terminus of the adeno associated virus Rep68/78 proteins. AB - Adeno-associated virus (AAV) replicates in the nucleus of infected cells, and therefore multiple nuclear import events are required for productive infection. We analyzed nuclear import of the viral Rep proteins and characterized a nuclear localization signal (NLS) in the C-terminus. We demonstrate that basic residues in this region constitute an NLS that is transferable and mediates interaction with the nuclear import receptor importin alpha in vitro. Mutant Rep proteins are predominantly cytoplasmic and are severely compromised for interactions with importin alpha, but retain their enzymatic functions in vitro. Interestingly, mutations of the NLS had significantly less effect on importin alpha interaction and replication in the context of Rep78 than when incorporated into the Rep68 protein. Together, our results demonstrate that a bipartite NLS exists in the shared part of Rep68 and Rep78, and suggest that an alternate entry mechanism may also contribute to nuclear localization of the Rep78 protein. PMID- 15351209 TI - Novel patterns of nevirapine resistance-associated mutations of human immunodeficiency virus type 1 in treatment-naive patients. AB - Several reports have recently shown that drug-resistant human immunodeficiency virus type 1 (HIV-1) is often isolated from treatment-naive patients. We phenotypically analyzed HIV-1 strains isolated from 44 treatment-naive individuals and found two strains highly resistant (69- and >310-fold) against nevirapine (NVP). Direct sequencing showed these two isolates had a novel mutation, K238S, in reverse transcriptase (RT), but did not have any reported NVP resistance-associated mutation. A 48-h culture in the presence of NVP, however, selected HIV-1 carrying NVP resistance-associated mutations, V106A, V108I, or both, suggesting that minor viral populations of these two isolates had harbored these mutations. Replication kinetic studies of recombinant HIV-1 clones suggested that K238S conferred a significant resistance against NVP, especially when accompanied with V106A (530-fold) or V108I (56-fold). Our study identified a novel NVP resistance-associated mutation, K238S, which could be persistently detected by genotypic assay longer than V106A and V108I during off-treatment period. PMID- 15351210 TI - Development of reassortant viruses between pathogenic hantavirus strains. AB - Segment reassortment of negative strand viruses is an important mechanism for the development of new virus strains with altered pathogenicity. This study reports on in vitro generation of reassortants between Andes (ANDV) and Sin Nombre (SNV) viruses. Although they both cause hantavirus pulmonary syndrome (HPS), ANDV is the only hantavirus that has been transmitted from person to person (). Following dual infection of cells with ANDV and SNV, 8.9% of 337 progeny plaques contained reassortants, of which 66% were diploid, and 34% were monoploid. The monoploid reassortants contained the S and L segments of SNV and ANDV M segment. Analysis of replication of the monoploid reassortant indicated its efficiency was similar to ANDV rather than SNV. Results described in this study illustrate the ability to rapidly generate new hantavirus genotypes between genetically unrelated viruses by gene reassortment and provide a tool to dissect the pathogenesis of these important viruses. PMID- 15351211 TI - Increased levels of B1 and B2 SINE transcripts in mouse fibroblast cells due to minute virus of mice infection. AB - Minute virus of mice (MVM), an autonomous parvovirus, has served as a model for understanding parvovirus infection including host cell response to infection. In this paper, we report the effect of MVM infection on host cell gene expression in mouse fibroblast cells (LA9 cells), analyzed by differential display. Somewhat surprisingly, our data reveal that few cellular protein-coding genes appear to be up- or downregulated and identify the murine B1 and B2 short interspersed element (SINE) transcripts as being increased upon MVM infection. Primer extension assays confirm the effect of MVM infection on SINE expression and demonstrate that both SINEs are upregulated in a roughly linear fashion throughout MVM infection. They also demonstrate that the SINE response was due to RNA polymerase III transcription and not contaminating DNA or RNA polymerase II transcription. Furthermore, expression of MVM NS1, the major nonstructural protein, by transient transfection also leads to an increase in both murine SINEs. We believe this is the first time that the B1 and B2 SINEs have been shown to be altered by viral infection and the first time parvovirus infection has been shown to increase SINE expression. The increase in SINE transcripts caused by MVM infection does not appear to be due to an increase in either of the basal transcription factors TFIIIC110 or 220, in contrast to that which has been shown for other viruses. PMID- 15351212 TI - Cross-reactive pseudovirus-neutralizing anti-envelope antibodies coexist with antibodies devoid of such activity in persistent hepatitis C virus infection. AB - Most RNA viruses have evolved mechanisms to avoid neutralizing antibody responses, and it is generally believed that variability of envelope-encoding regions is the major molecular basis of this phenomenon. However, it has been hypothesized that other mechanisms can be involved. Recent experimental data indicate that in hepatitis C virus (HCV) infection, the anti-envelope humoral response includes cross-reactive antibody clones able to neutralize vesicular stomatitis virus (VSV) pseudotypes containing HCV E1 and E2 glycoproteins (HCV/VSV pseudotype) as well as other clones devoid of such activity. In this work, we demonstrate that natural infection with a large variety of HCV isolates belonging to different genotypes elicits HCV/VSV pseudotype-neutralizing cross reactive anti-envelope antibodies together with clones unable to neutralize this pseudovirus. This was shown by designing a novel strategy for quantitation of serum antibodies binding selectively to single viral cross-reactive conformational epitopes. These data can be useful not only for a better understanding of the virus-host interplay in important viral diseases, but also for the development of an effective anti-HCV vaccine. PMID- 15351213 TI - Nuclear localization of Vpr is crucial for the efficient replication of HIV-1 in primary CD4+ T cells. AB - The human immunodeficiency virus type 1 (HIV-1) accessory protein Vpr appears to make a substantial contribution to the replication of HIV-1 in established T cell lines when HIV-1 is present at very low multiplicities of infection. However, the role of Vpr in viral replication in primary CD4+ T cells remains to be clarified. In this study, we generated a panel of viruses that encoded mutant forms of Vpr that lacked either the ability to accumulate in the nucleus and induce G2 arrest or the ability to induce apoptosis, which has been shown to occur independently of G2 arrest of the cell cycle. We demonstrate here that the nuclear localization of Vpr and consequent G2 arrest but not the induction of apoptosis by Vpr are important for viral replication in primary CD4+ T cells at both high and low multiplicities of infection. Viruses that encoded mutant forms of Vpr that failed to be imported into the nucleus in the presence of cytoplasmic extracts from primary CD4+ T cells in an in vitro nuclear import assay replicated at drastically reduced rates. Thus, Vpr might be a key regulator of the viral nuclear import process during infection in primary CD4+ T cells. By contrast, a mutant form of Vpr that exhibited diffuse cytosolic staining exclusively in an immunofluorescence assay of HeLa cells and was not imported into nucleus by the cytosol from HeLa cells was effectively imported into the nucleus by cytosol from primary CD4+ T cells. This Vpr mutant virus replicated well in primary CD4+ T cells, indicating that cellular factors in primary CD4+ T cells are indispensable for the accumulation of Vpr in the nucleus and, thus, for viral replication. Our results suggest that the nuclear import of Vpr might be a good target in efforts to block the early stages of replication of HIV-1. PMID- 15351214 TI - Characterization of a DNA vaccine expressing a human immunodeficiency virus-like particle. AB - An ideal human immunodeficiency virus type-1 (HIV-1) vaccine will most likely need to elicit cross-reactive neutralizing antibodies and a strong cell-mediated immune response against multiple HIV-1 antigens to confer protection against challenge. In this study, DNA vaccines were constructed to express virally regulated human immunodeficiency virus-like particles (VLP) to elicit broad spectrum immune responses to multiple HIV-1 antigens. VLPs were efficiently produced using sequences encoding gag and pol gene products from an X4 isolate and sequences encoding for tat, rev, vpu, and env from R5 or R5X4 isolates. The integrase, vpr, vif, and nef genes were deleted. In addition, the long terminal repeats (LTRs) were removed and transcription of the VLP insert was driven by the addition of the cytomegalovirus immediate-early (CMV-IE) promoter. A second generation of VLP vaccine plasmids was constructed with mutations engineered into the VLP DNA to produce particles deficient in activities associated with viral reverse transcriptase and protease. Primate cell lines, transiently transfected with DNA, efficiently secreted VLP into the supernatant that banded within a sucrose gradient at densities similar to infectious virions. In addition, these particles incorporated Env on the particle surface that bound soluble human CD4. These VLPs provide a safe and efficient strategy for presenting multiple HIV-1 antigens, expressed from a single insert, to the immune system in a structure that mimics the infectious virion. PMID- 15351215 TI - In vivo mutational analysis of the N-terminal region of HIV-1 Nef reveals critical motifs for the development of an AIDS-like disease in CD4C/HIV transgenic mice. AB - HIV-1 Nef is a critical determinant of pathogenicity in humans and transgenic (Tg) mice. To gain a better understanding of the molecular mechanisms by which Nef induces an AIDS-like disease in Tg mice, a mutational analysis of the N terminal domain, involved in anchoring Nef to the plasma membrane, was carried out. The pathogenic effects of these Nef mutant alleles were evaluated in Tg mice by FACS analysis and by histopathological assessment. Mutation of the myristoylation site (G2A) completely abrogated the development of the AIDS-like organ disease in Tg mice, although partial downregulation of the CD4 cell surface protein and depletion of peripheral CD4+ T-cells, but not of CD4(+)CD8+ thymocytes, still occurred. Despite that, the peripheral CD4+ T cells expressing Nef(G2A) show normal spontaneous proliferation in vivo or after stimulation in vitro, including in an allogenic mixed leukocyte reaction (MLR). Three other internal deletion mutants of Nef, spanning amino acids 8-17 (Nef(Delta8-17)), 25 35 (Nef(Delta25-35)), and 57-66 (Nef(Delta57-66)), were also studied. Nef(Delta8 17) retained full pathogenic potential, although Nef(Delta25-35) and Nef(Delta57 66) Tg mice were free of organ disease. However, Nef(Delta25-35) Tg mice exhibited disorganization of thymic architecture and a partial depletion of peripheral CD4+ T cells. These data indicate that myristoylation and other regions at the N-terminus of Nef (aa 25-35 and 57-66) are involved in mediating severe T-cell phenotypes and organ disease, although residues 8-17 are dispensable for these Nef functions. In addition, these results indicate that at least some of the CD4+ T-cell phenotypes can develop independently of the other AIDS-like organ phenotypes. This apparent segregation of different Nef-mediated phenotypes suggests distinct mechanisms of Nef action in different populations of target cells, and may be relevant to human AIDS. PMID- 15351216 TI - Mutations that promote furin-independent growth of Semliki Forest virus affect p62-E1 interactions and membrane fusion. AB - The enveloped alphavirus Semliki Forest virus (SFV) infects cells via a low pH triggered membrane fusion reaction mediated by the E1 protein. E1's fusion activity is regulated by its heterodimeric interaction with a companion membrane protein E2. Mature E2 protein is generated by furin processing of the precursor p62. Processing destabilizes the heterodimer, allowing dissociation at acidic pH, E1 conformational changes, and membrane fusion. We used a furin-deficient cell line, FD11, to select for SFV mutants that show increased growth in the absence of p62 processing. We isolated and characterized 7 such pci mutants (p62 cleavage independent), which retained the parental furin cleavage site but showed significant increases in their ability to carry out membrane fusion in the p62 form. Sequence analysis of the pci mutants identified mutations primarily on the E2 protein, and suggested sites important in the interaction of p62 with E1 and the regulation of fusion. PMID- 15351217 TI - The phosphoprotein (P) and L binding sites reside in the N-terminus of the L subunit of the measles virus RNA polymerase. AB - Measles virus encodes an RNA-dependent RNA polymerase composed of the L and P proteins. Recent studies have shown that the L proteins of both Sendai virus and parainfluenza virus 3 form an L-L complex [Cevik, B., Smallwood, S., Moyer, S.A., 2003. The oligomerization domain resides at the very Nterminus of the Sendai virus L RNA polymerase protein. Virology 313, 525-536.; Smallwood, S., Moyer, S.A., 2004. The L polymerase protein of parainfluenza virus 3 forms anoligomer and can interact with the heterologous Sendai virus L, P and C proteins. Virology 318, 439-450.; Smallwood, S., Cevik, B., Moyer, S.A., 2002. Intragenic complementation and oligomerization of the L subunit of the Sendai virus RNA polymerase. Virology 304, 235-245.]. Using differentially tagged L proteins, we show here that measles L also forms an oligomer and the L-L binding site resides in the N-terminal 408 amino acids overlapping the P binding site in the same region of L. To identify amino acids important for binding P and L, site-directed mutagenesis of the L-408 protein was performed. Seven of twelve mutants in L-408 were unable to form a complex with measles P while the remainder did bind at least some P. In contrast, all of the mutants retained the ability to form the L L complex, so different amino acids are involved in the L and P binding sites on L. Four of the 408 mutations defective in P binding were inserted into the full length measles L protein and all retained L-L complex formation, but did not bind P. Full-length L mutants that did not bind P were also inactive in viral RNA synthesis, showing a direct correlation between P-L complex formation and activity. PMID- 15351218 TI - The increased replicative capacity of a late-stage simian immunodeficiency virus mne variant is evident in macrophage- or dendritic cell-T-cell cocultures. AB - Human and simian immunodeficiency virus (HIV and SIV) may co-opt antigen capture and presentation functions of antigen presenting cells (APCs) to facilitate infection of CD4+ T-cells. To address whether the replicative capacity of SIV in the host may be associated with the extent of viral replication in response to APC-T-cell interactions, we compared the replicative phenotypes of cloned early and late-stage SIVmne variants of known pathogenicity. Here, we show that the highly pathogenic late variant SIVmne027 replicates more efficiently in both macrophage- and dendritic cell (DC)-T-cell cocultures than the minimally pathogenic early virus SIVmneCl8. Contact between either macrophages or DC and T cells increases replication of SIVmne027. Our analysis also demonstrates that mutations in pol and nef contribute to the greater replicative capacity of SIVmne027 in DC- or macrophage-T-cell cocultures. Together, these data suggest that variant viruses that evolve to replicate vigorously in response to APC-T cell interactions may have increased replicative capacity in vivo. PMID- 15351220 TI - The UK Public Health Association and Congress of the World Federation of Public Health Associations (WFPHA) conference-plenary session presentations. PMID- 15351221 TI - Suburban sprawl and physical and mental health. AB - OBJECTIVE: To study the association between objective measures of suburban sprawl and chronic medical conditions and mental health disorders in the USA. METHODS: Cross-sectional analysis of survey data merged with objective measures of suburban sprawl. Outcomes are self-reported medical conditions, mental health disorders and health-related quality of life. RESULTS: Sprawl significantly predicts chronic medical conditions and health-related quality of life, but not mental health disorders. An increase in sprawl from one standard deviation less to one standard deviation more than average implies 96 more chronic medical problems per 1000 residents, which is approximately similar to an aging of the population of 4 years. CONCLUSIONS: A robust association between sprawl and physical (but not mental) health suggests that suburban design may be an important new avenue for health promotion and disease prevention. PMID- 15351222 TI - Mechanisms by which area-based regeneration programmes might impact on community health: a case study of the new deal for communities initiative. AB - STUDY OBJECTIVE: To explore mechanisms by which area-based regeneration initiatives may impact on health. DESIGN: Case study of the New Deal for Communities programme, part of the UK National Strategy for Neighbourhood Renewal. MAIN RESULTS: Health impacts may result from three mechanisms: sociospatial stigma, community participation and the commissioning of projects designed to change the distribution of determinants of health, including access to services and healthy lifestyles. CONCLUSIONS: The present UK National Strategy for Neighbourhood Renewal includes improving health status as one of its key outcome measures. Area-based regeneration schemes such as the New Deal for Communities initiative should, intuitively, bring about health gain through their participatory approach to changing the physical and socio-economic environments. At present, however, the direction and magnitudes of these impacts are not clear. PMID- 15351223 TI - Epidemiology of spider bites in Texas, 1998-2002. PMID- 15351224 TI - Response patterns to a postal survey using a cervical screening register as the sampling frame. AB - This study compared different types of respondent to a postal survey. A random sample of women aged 20-64 years (n = 4057) was selected from a population-based cervical screening register to examine their information experience during the screening programme. The initial response rate was 57%, and this increased to 81% after one reminder. Respondents were older (P < 0.0001) than non-respondents, but both groups were comparable with regard to attendance history for cervical screening (respondents 89.3%; non-respondents 89.1%) and normal smear test results (respondents 84.0%; non-respondents 81.4%). Early and late respondents were similar in age. Compared with late respondents, early respondents were more likely to live in highly affluent (P < 0.0001) and rural areas (P = 0.026). They were also more likely to be non-attenders (P < 0.0001), but they were less likely to have had inadequate smear results (P = 0.030) than late respondents. These results suggest that consideration should be given to factors other than sociodemographic differences when examining response patterns to postal surveys. PMID- 15351225 TI - Breast cancer incidence and its possible spatial association with pesticide application in two counties of England. AB - BACKGROUND: This study examines the spatial distribution of breast cancer incidence in Lincolnshire and Leicestershire, and its association with the application of selected pesticides. METHODS: The Black-White (BW) join-count statistic and Moran I coefficient were used to investigate the localized distribution of breast cancer. Linear regression techniques were applied to examine the association between the breast cancer incidence rate and pesticide application. RESULTS: The results from the BW join-count test and the Moran I coefficient test showed no overall evidence of localized distribution, 'clusters', of breast cancer. The regression analyses showed no sign of spatial association between breast cancer and application of pesticides in the urban areas as expected. The findings, however, did reveal a spatial association between the breast cancer incidence rates and the application of three selected pesticides [Aldicarb, Atrazine and Lindane (thought to potentially mimic oestrogen)] in rural Leicestershire. No such association was seen in Lincolnshire. CONCLUSIONS: Pesticides vary significantly in their spatial application. Overall, consistent associations between breast cancer incidence rates and the selected pesticides were not found, although the ecological study design has limitations and these are discussed. This approach is able to rule out strong associations; assessment of smaller risk would require a large and expensive study in individuals. The results of this study, although derived in the UK, have significance to the debate concerning the possibility of environmental compounds that mimic oestrogen and their consequences for human health. PMID- 15351226 TI - Influence of physicians' attitudes on under-notifying infectious diseases: a longitudinal study. AB - OBJECTIVES: To identify practitioners' demographic and professional characteristics associated with reporting of mandatory-reporting diseases (MRDs), and to identify attitudes associated with MRD reporting. DESIGN: Longitudinal study. SETTING: Regional Health Service of Galicia (North-western Spain). SUBJECTS: Random sample of 600 physicians. MAIN OUTCOME MEASURES: A postal questionnaire was used to measure the physicians' beliefs, knowledge and attitudes regarding MRDs. Associations between the independent variables and outcomes (notification or non-notification of MRDs every week during 1998) were modelled using the Andersen-Gill proportional hazards model. RESULTS: The response rate was 60.1%. The following beliefs, knowledge and attitudes were statistically associated with a smaller probability of reporting any given MRD: (1) I would only notify an MRD after confirming diagnosis; (2) the MRD reporting system interferes with my daily clinical practice; (3) besides the legal requirements, I would have to report MRDs as a medical professional; and (4) only the reporting of relevant or severe MRDs is necessary. Under-reporting was not associated with specialization (general or paediatric) or the type of contract (fixed or temporary), but was associated with gender. CONCLUSIONS: Some physicians' beliefs, knowledge and attitudes regarding MRDs are associated with under-reporting. This suggests that modification of certain attitudes and knowledge in physicians could greatly reduce the under-reporting of MRDs. PMID- 15351227 TI - Doctors, patients and influenza-like illness: clinicians or patients at risk? AB - BACKGROUND: Healthcare providers can transmit influenza and influenza-like illness (ILI) to patients and vice versa. However, the magnitude of this problem in the healthcare system as a whole is unknown. Using population-based administrative health data, we tested the hypothesis of a temporal association of ILI diagnosis among clinicians and their patients. METHODS: Healthcare providers under study included physicians (85%) and a variety of other medical professionals (such as chiropractors, dentists and optometrists). Cases of ILI were defined as having an ICD-9 code of influenza (487), pneumonia (480-486) or bronchitis (466 and 490) in a province-wide healthcare-provider billing system. Rates of ILI among persons who saw a sick (case) and non-sick (control) doctor were calculated and compared. RESULTS: The rate of ILI was lower among providers than among patients for every year of the study. The mean number of exposures to patients diagnosed with ILI was higher among case providers than among control providers (P = 0.044). However, exposure to case providers did not significantly increase the risk of ILI diagnosis among patients (OR=1.11; 95% confidence intervals 0.85-1.36). INTERPRETATION: Our findings are consistent with the hypothesis of patient-to-provider transmission. However, we may have underestimated the association and magnitude of this effect. Our results do not support the hypothesis that infectious doctors transmit disease to their patients, although this may be partly accounted for by the lower rates of ILI among the provider population in general. PMID- 15351228 TI - Correlates of heavy backpack use by elementary school children. PMID- 15351229 TI - Marasmius oreades lectin induces renal thrombotic microangiopathic lesions. AB - The present studies demonstrate that infusion of a type B specific lectin derived from the mushroom Marasmius oreades (MOA) into mice binds selectively to the glomerular endothelial cells via surface carbohydrate moieties resulting in cell injury and death associated with platelet-fibrin thrombi. This selective MOA binding to the endothelial cells can be abrogated by a sugar specific for the carbohydrate sequence. Hemolytic-Uremic Syndrome (HUS) and the closely associated Thrombotic Thrombocytopenic Purpura (TTP) are diseases associated with widespread microvascular injury in various organs. Clinically, these diseases are associated with microangiopathic hemolytic anemia and thrombocytopenia. The kidney glomerulus is a primary target of this microvascular injury. There are many underlying etiologies including bacterial toxins. Experimentally, such toxins injure endothelial cells in vitro but in vivo studies have failed to reproduce the characteristic renal pathology. We suggest that MOA-induced glomerular microangiopathic injury could be used to study the pathophysiology of endothelial cell injury as related to glomerular microangiopathic injury. PMID- 15351230 TI - Prevalence of MTHFR gene polymorphisms (C677T and A1298C) among Tamilians. AB - We have investigated the incidence of the C677T and A1298C methylene tetrahydrofolate reductase (MTHFR) gene single nucleotide polymorphisms (SNPs) in the South Indian Tamil Nadu population with a total number of 72 individuals. The MTHFR genotyping was performed using the polymerase chain reaction followed by restriction enzyme analysis. Homozygosity for the MTHFR A1298C SNP was detected in 15.3% (11/72) of the individuals tested, and 47.2% (34/72) were heterozygous for this SNP. Homozygosity for the C677T MTHFR SNP was detected in 1.38%(1/72), and the frequency of the C677T heterozygotes was 18.1%(13/72). When we analyzed the combined frequency of the two SNPs, the frequency of double heterozygosity was19.6%, and the frequency of double homozygosity was completely absent among the study group. The 'C' allele frequency for MTHFR A1298C was 0.389, and the 'T' allele frequency for C677T mutation was 0.104. Out of the 72 individuals included in the study, 52 were acute myocardial infarction (AMI) patients and 20 were healthy individuals with no documented history of heart disease. The results of this study indicate that the MTHFR A1298C SNP is more prevalent among the Tamilians when compared to the MTHFR C677T SNP, suggesting a possible role of MTHFR A1298C in the pathogenesis of heart diseases. PMID- 15351231 TI - Enhancing sensitivity of human herpes virus diagnosis with DNA microarrays using dendrimers. AB - DNA microarray technology has become a promising new tool for the detection and identification of viral pathogens in human plasma and cell cultures. For exploration of this technology, we have developed DNA microarrays that encode capture oligonucleotide probes for different human herpes viruses: herpes simplex virus (HSV) HSV-1, HSV-2, varicella zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), and HHV-6. The on-chip hybridization is accomplished with the PCR amplicons of the respective human herpes virus types. In this original article, we attached multiple Cy3-fluorophores to the branched 5' ends of the labeling oligonucleotide primers. For the first time, we experimentally demonstrated that the self-designed, knowledge-based, and focused microarrays specifically hybridized to fluorophore-labeled pathogenic DNAs using dendrimer technology. The fluorescence signal enhancement via the dendrimers was up to 30 times compared with the quenched single Cy3-fluorophore-labeled HSV-1 DNA. The on chip signal-amplifying effect depended upon the number of branches and the concentration of fluorophore-labeled pathogenic DNAs. Treblers were superior to doublers, as trebler-labeled nucleic acids had fluorescence-signal-enhancing effects over a broad range of labeled DNA concentrations exemplified for the quenched single Cy3-fluorophore-labeled HSV-1 and non-quenched single Cy3 fluorophore-labeled CMV DNAs. PMID- 15351232 TI - RNA degradation in human breast tissue after surgical removal: a time-course study. AB - There is much interest in the study of human malignancy using gene expression profiling techniques. Expression profiles obtained from microarrays utilize RNA extracted from the tissue in question. Currently, cell cultures or fresh tissue processed "quickly" are used in these studies. To our knowledge, there are no published reports of a time-course of RNA degradation in surgically removed breast tissue. Such a time-course study is critically needed. We obtained normal breast tissue from breast reduction surgery. Portions of breast tissue kept at room temperature were sampled and placed into RNAlater to preserve RNA at different time-points from 10 min to 3 h after the surgical removal. We evaluated total RNA integrity from each specimen using agarose gel electrophoresis and real time quantitative RT-PCR analysis of four genes. Electrophoresis showed good quality, intact RNA at all time points up to 3 h. Quantitative RT-PCR showed no difference in amplified products among all samples. Our study showed that there was no loss of RNA integrity in normal breast tissue for up to 3 h after surgical removal. PMID- 15351233 TI - Functional and structural characterization of two populations of human monocyte derived dendritic cells. AB - BACKGROUND: The characterization of human monocyte-derived dendritic cells (HM DC) subsets have been a very difficult and elusive task because of the lack of appropriate reagents. We, therefore utilized several diverse approaches to evaluate two populations of HM-DC including flow cytometry, ultra-structural evaluation by electron microscopy, and functional assays. In addition, we studied the kinetics of the expression of antigens on HM-DC at diverse intervals of time and identify surface markers and functional differences of these two HM-DC subsets. RESULTS: This study identified that a phenotype of HM-DC as defined by CD11c+, CD86+, and CD40+ could be separated in the presence or absence of TGF beta1 into two different subsets of DC: (i) HM-DC without Birbeck granuli (Mo-DC) and (ii) HM-DC with Birbeck granuli (Mo-LC). Furthermore, the functional studies showed that the HM-DC treated with TGF-beta1 (Mo-LC) exhibited the presence of Birbeck granuli and could actively divide. In addition, after undergoing more than four cell divisions, these cells split into at least two additional subsets of Mo-LC: (iia) Mo-LC with high forward scatter (FSC) and (iib) Mo-LC with normal FSC. In contrast, the Mo-DC cultured in absence of TGF-beta1 did not exhibit Birbeck granuli, showed reduced ability to divide, and kept the normal FSC when analyzed. CONCLUSIONS: This study enabled us to determine in HM-DC: (i) the existence of antigenic and functional differences between various subpopulations of Mo-DC and Mo-LC; (ii) the existence of differences in the kinetics of antigens expression among the subsets of Mo-DC and Mo-LC; (iii) the existence of specific markers for each of the subpopulations of HM-DC. PMID- 15351234 TI - Modulation of thyroidal radioiodine uptake by theophylline. AB - Diagnostic and therapeutic use of radioiodine in the management of thyroid disorders depends on the ability of thyroid cells to concentrate radioiodine, a process regulated by thyrotropin and dependent on the intracellular increase in cAMP. We tested the ability of theophylline, a drug known to increase intracellular cAMP via inhibition of phosphodiesterase, to modulate the thyroidal radioiodine uptake in FRTL-5 cells, in mice and in humans. In FRTL-5 cells, theophylline increased the uptake of radioactive iodine and intracellular cAMP only at low concentrations (1 microM). In mice, theophylline increased slightly the radioiodine uptake, although this increase varied from 1.5- to 6.6-fold. In humans, theophylline decreased slightly the radioiodine uptake, a decrease that became more pronounced with time after radioiodine administration. These studies suggest that theophylline modulates the radioiodine uptake in a dose-dependent fashion, although the modulation is mild and probably not applicable to the clinical setting. PMID- 15351235 TI - Oxidative stress in viral hepatitis and AIDS. AB - Dominant types of viral hepatitis are presently A, B, and C with prophylactic immunization available only for A and B. Hepatitis B and C and human immunodeficiency virus (HIV) infection constitute a worldwide scourge and treatment is far from satisfactory. Each produces severe oxidative stress (OS) and secondary cellular damage of varying severity and, as in toxic hepatitis, progression and regression are dependent on redox balance between oxidation and antioxidation. Experimental and clinical studies suggest that xenobiotics and co infections exert cumulative, detrimental effects on their pathogeneses and further deplete antioxidants. It is proposed therefore that in the clinical management of these infections and especially in their early stages, considerable benefit should accrue from antioxidant repletion at dosages substantially above recommended daily allowances (RDAs) in conjunction with a nutritious high protein diet. Because plasma zinc and selenium concentrations are very low, their replenishment by high dosages is urgent and mandatory particularly in advanced HIV infections bordering on acrodermatitis enteropathica. Also recommended is their long-term continuance at high normal levels. PMID- 15351236 TI - Experimental encephalomyocarditis virus infection in pregnant mice. AB - The present study was carried out to clarify the mode of encephalomyocarditis (EMC) virus infection in pregnant mice. Pregnant BALB/c mice were intraperitoneally inoculated with the D variant of EMC virus (EMC-D) (5 x 10(2) PFU/mouse) on 11 days of gestation and killed at 1, 3, and 5 days post inoculation (DPI). The virus titer (dam's serum, placenta, and fetus), histopathology (fetus, placenta, and uterus), distribution of viral RNA (fetus, placenta and uterus), and ultrastructure (fetal heart and placenta) were examined. No deaths occurred to fetuses at 1 DPI but almost all fetuses died at 5 DPI. The virus titers of dam's serum and placenta were elevated at 1 DPI, peaked at 3 DPI, and the former was not detected at 5 DPI. The virus titer of fetus was first elevated at 3 DPI and the level was lower than those of others. Histopathological changes and signals of viral RNAs detected by in situ hybridization (ISH) were observed in the spongiotrophoblast layer of placenta and in the fetal myocardium and liver at 3 DPI. The uterus was free from lesions and signals of viral RNA. Ultrastructural changes developed in trophoblast cells and giant cells in the spongiotrophoblast layer at and after 1 DPI and in fetal myocardial cells at 3 DPI. In the cytoplasm of trophoblast cells and giant cells, aggregations of virus-like particles 20-30 nm in diameter were observed in crystalline array. These results suggest that trophoblast cells and giant cells in the spongiotrophoblast layer are the main target of EMC virus in the placenta and that placental damage as well as the direct effect of virus to fetuses may be a cause of fetal death. PMID- 15351237 TI - Ureaplasma urealyticum binds mannose-binding lectin. AB - Mannose-binding C-type lectin (MBL) is an important component of innate immunity in mammals. Mannose-binding lectin (MBL), an acute phase protein, acts as an opsonin for phagocytosis and also activates the mannan-binding lectin complement pathway. It may play a particularly significant role during infancy before adequate specific protection can be provided by the adaptive immune system. Ureaplasma urealyticum has been linked to several diseases including pneumonia and chronic lung disease (CLD) in premature infants. We therefore investigated the ability of U. urealyticum to bind MBL. A guinea pig IgG anti-rabbit-MBL antiserum was produced. An immunoblot (dot-blot) assay done on nitrocellulose membrane determined that the anti-MBL antibody had specificity against both rabbit and human MBL. Pure cultures of U. urealyticum, serotype 3, were used to make slide preparations. The slides containing the organisms were then incubated with nonimmune rabbit serum containing MBL. Ureaplasma was shown to bind rabbit MBL with an immunocytochemical assay using the guinea pig IgG anti-rabbit MBL antiserum. Horseradish peroxidase (HRP)-labeled anti-guinea pig IgG was used to localize the reaction. The anti-MBL antiserum was also used in an immunocytochemical assay to localize U. urealyticum in histological sections of lungs from mice specifically infected with this organism. The same method also indicated binding of MBL by ureaplasma in human lung tissue obtained at autopsy from culture positive infants. Our results demonstrate that ureaplasma has the capacity to bind MBL. The absence of MBL may play a role in the predisposition of diseases related to this organism. PMID- 15351238 TI - Localization of vascular endothelial growth factor during the early reparative phase of the rats' vessels deprivation-induced osteonecrosis of the femoral heads. AB - The expedited revascularization of the rats' avascular, necrotic femoral heads suggests the operation of angiogenic factor(s). The blood circulation of the epiphysis was interrupted by cutting the cervical periosteum and the ligamentum teres of rats' femoral heads. Three days postoperatively, the marrow was necrotic. Seven days postoperatively, the subchondral bony plate and trabecular bone were necrotic as well. The joint capsule was distended by myriad, so-called synovial fibroblasts, all of which were virtually immunoreactive with an antibody to vascular endothelial growth factor. The expression of this factor in the synovial membrane of non-operated rats was limited to preexisting blood vessels. Revascularization of necrotic, avascular femoral heads makes up the essential step in the chain of events terminating in the repair processes, that is, resorption of the necrotic debris and its substitution by newly formed bony and hematopoietic-fatty tissues. Synthesis and release of excessive amounts of vascular endothelial growth factor by these fibroblasts explain the lively angiogenesis in the necrotic intertrabecular spaces of the femoral heads. PMID- 15351239 TI - Development of early apoptosis and changes in T-cell subsets in mouse thymocyte primary cultures treated with nivalenol. AB - Nivalenol (NIV), a trichothecene mycotoxin, is a secondary fungal metabolite mainly produced by Fusarium nivale. We first reported that NIV could induce apoptosis and changes in lymphocyte subsets in lymphoid tissues of mice. In this study, to clarify the direct effects of NIV on thymocytes, mouse thymocyte primary cultures were treated with NIV at the dose levels of 0.25, 0.5 and 1.0 microg/ml and examined for up to 24 h after treatment by flow cytometry. The number of viable cells decreased significantly at and after 6 h in dose- and time dependent manners, and FACS analysis revealed that the apoptotic cell index showed a significant increase in all treated groups at and after 3 h in a time dependent manner. The index at 24 h was lowest in 1.0 microg/ml-group. The number of CD4(+)CD8(+) cells was prominently depleted in all groups in a time-dependent manner. On the other hand, the numbers of CD4(+)CD8(-) and CD4(-)CD8(+) cells were significantly depleted only in 1.0 microg/ml-group at 24 HAT. These results indicate that NIV directly affects thymocytes and induces apoptosis mainly in CD4(+)CD8(+)cells. PMID- 15351241 TI - Adult traumatic brachial plexus injury. AB - Injury to the brachial plexus in the adult is usually a closed injury and the result of considerable traction to the shoulder. Brachial plexus injury in the adult is an increasingly common clinical problem. Recent advances in neurosurgical techniques have improved the outlook for patients with brachial plexus injuries. The choice of surgical procedure depends on the level of the injury and the radiologist has an important role in guiding the surgeon to the site of injury. This article will describe the anatomy and pathophysiology of traction brachial plexus injury in the adult. The neurosurgical options available will be described with emphasis on the information that the surgeon wants from imaging studies of the brachial plexus. The relative merits of MRI and CT myelography are discussed. PMID- 15351240 TI - Microvessel density as a molecular marker for identifying high-grade prostatic intraepithelial neoplasia precursors to prostate cancer. AB - BACKGROUND: Existing clinical data have shown that high-grade prostatic intraepithelial neoplasia (HGPIN) is the most likely precursor to prostate cancer (CaP). Criteria to distinguish HGPIN that progress to CaP from those that do not remain poorly defined. Our objective was to evaluate microvessel density as a molecular marker for distinguishing HGPINs that have the potential of progressing to cancer. MATERIALS AND METHODS: Human prostatic tissue samples were collected randomly from 50 prostatectomy and cystoprostatectomy patients. Formalin-fixed and paraffin-embedded sections were used for immunohistochemical localization of rabbit anti-human von Willebrand factor VIII (vWF) IgG, mouse anti-high molecular weight cytokeratin 34BE-12 in basal cells, and mouse anti-heparan sulphate proteoglycan (HSPG) IgGs in basement membranes associated with benign prostatic hyperplasia (BPH), PIN associated with some BPH (isolated PIN), and PIN associated with CaP. RESULTS: Analysis of immunostaining data showed that PINs could be categorized according to their distributions within and outside 2 standard deviations (SD) of the mean for microvessel density. The average number of microvessels was significantly higher (P < 0.0001) in PINs associated with Gleason score 7 tumors than those associated with Gleason scores 4-6 (P < 0.1328) or 8 and 9 tumors (P < 0.1708). Morphologically, PINs within 2 SD were composed of low- and high-grade type, whereas those outside 2 SD of microvessel density were predominantly of high-grade type. Cytokeratin and HSPG localization patterns also showed differences in PINs found within and outside 2 SD of microvessel density. We found localization of cytokeratin 34BE-12 in basal cells of specimens with BPH alone, isolated PIN, and PIN associated with CaP within 2 SD, whereas many PINs outside 2 SD showed disruptions in cytokeratin localization. The basement membranes of PINs within 2 SD of microvessel density were relatively intact, whereas those outside 2 SD were fragmented. CONCLUSIONS: Our immunostaining data indicates that once HGPIN is found in the initial prostatic biopsy, it should be evaluated for microvessel density by localization of vWF. Our data indicate that characteristics of HGPIN can be augmented by evaluations of cytokeratin and HSPG molecular markers to assess the potential of HGPIN progression to malignancy. When biopsy samples show HGPIN with increased microvessel density and disrupted cytokeratin and HSPG markers, the patient may be a candidate for repeat biopsy. Since our study is limited to 50 prostate tissue samples, we emphasize that our conclusion is tentative and ought to be confirmed in a study with a larger sample size. This is the first report to show that microvessel density may distinguish HGPIN that is a precursor to prostate cancer. PMID- 15351242 TI - PET/CT "Cometh the hour, cometh the machine?". PMID- 15351243 TI - Splenic abnormalities: a comparative review of ultrasound, microbubble-enhanced ultrasound and computed tomography. AB - The ultrasound appearances of abnormalities of the spleen are reviewed and images compared with computed tomography. Focal lesions, both benign and malignant, trauma, infarction and congenital abnormalities are presented. The use of microbubble ultrasound contrast media as an aid to identifying and characterizing abnormalities is discussed. PMID- 15351244 TI - Contour irregularities of the distal femur caused by developmental, traumatic, and benign cortically-based neoplastic conditions: radiographic and MRI correlation. AB - This review illustrates the radiographic and magnetic resonance imaging (MRI) appearances of a variety of entities that result in surface contour abnormalities of the distal femur. Such abnormalities may result from developmental variants expressed in the growing skeleton, from trauma, or from cortically-based tumours. As the number of MRI examinations of the knee performed annually to exclude internal derangement increases, the frequency with which these variants and pathological entities are encountered will likewise increase. In most cases, the location of the lesion, as well as its specific imaging characteristics, will allow an accurate diagnosis. PMID- 15351245 TI - Radiofrequency ablation of bony metastatic disease. AB - Image-guided radiofrequency (RF) ablation is an emerging alternative therapy for treatment of cancer in patients who can not undergo surgical resection. We have performed RF ablation on patients to locally treat and palliate painful osseous metastatic disease for several years. We present an overview of various applications of RF ablation for treating osseous metastatic disease. PMID- 15351246 TI - Percutaneous radiofrequency ablation for unresectable large hepatic tumours during hepatic blood flow occlusion in four patients. AB - AIM: To evaluate percutaneous radiofrequency (RF) ablation therapy for unresectable large hepatic tumours combined with regional interruption of hepatic blood flow, and to assess the safety and efficacy of this procedure. MATERIALS AND METHODS: Four patients with hepatic tumours were enrolled in this study. Patients were treated by a single session of RF ablation during occlusion of both hepatic artery and hepatic vein. Tumour size ranged from 45-57 mm (mean 50.2 mm). Initial therapeutic efficacy was evaluated with helical computed tomography (CT) performed within 9 days after the treatment. CT or magnetic resonance imaging (MRI) was performed every 2-3 months thereafter. RESULTS: The largest axis of coagulated lesions after the ablation was 50-60 mm (mean 56.5 mm) in diameter. The ablation therapy was considered complete in three patients; after a mean follow-up of 12.7 months, CT and MRI revealed complete destruction of their tumours. One patient required further treatment. No severe complications occurred. CONCLUSION: Although further studies are needed, in this limited clinical trial a local ablation area exceeding 50 mm in diameter was achieved safely. PMID- 15351247 TI - Helical CTPA in the investigation of pulmonary embolism: a 6-year review. AB - AIM: To assess the change in practice and resulting diagnostic outcome in the radiological investigation of pulmonary embolism (PE) since the introduction of helical CTPA in a large teaching hospital. MATERIALS AND METHODS: A retrospective review was undertaken of all radiological investigations performed over a 6-year period before and after the introduction of CTPA (protocols 1 and 2, respectively) as an integral part of the imaging protocol in the investigation of clinically suspected PE. The total numbers and results of all investigations are assessed for each protocol. RESULTS: A substantial increase in both the total number of patients and the number of investigations performed for the investigation of PE since the introduction of CTPA occurred. Five hundred and twenty-six patients underwent 617 investigations in 1995-1996, and 760 patients underwent 805 investigations in 2001-2002. There was a significant decrease in the number of investigations per patient, 1.17 in 1995-1996 versus 1.06 in 2001 2002. Primary investigation showed a significant decrease in indeterminate examinations from 25.7 to 8.5% and an increase in positive results for PE from 18 to 24%. CONCLUSION: In the study population there was a significant increase in the number of patients being investigated for PE, with a decrease in both the number of non-diagnostic investigations and the total investigations per patient since the introduction of helical CTPA. This is probably due to the ready availability of a new imaging technique and physicians awareness that CTPA has significantly improved specificity, which encourages the referral of patients for investigation. PMID- 15351248 TI - Imaging of pleural fluid in healthy individuals. AB - AIM: The aim of the present study was to determine the ability of radiography and ultrasonography to detect normal pleural fluid in healthy individuals and to assess the frequency of this finding. MATERIALS AND METHODS: Chest ultrasonography of both pleural spaces was performed in a group of 106 healthy volunteers to identify pleural fluid, first in the lateral decubitus position and than leaning on one elbow. Posteroanterior (PA) and lateral decubitus expiratory radiography were subsequently performed. An anechoic layer at least 2 mm thick on chest ultrasonography and a density with a horizontal level at least 3 mm in depth on lateral decubitus radiography were taken as positive results. RESULTS: On ultrasonography the fluid layer with a typical wedge-shaped appearance was visible in the pleural space of 28 of 106 (26%) volunteers, on both sides in 17 of 28 (61%) and unilaterally in 11 of 28 (39%). The mean fluid layer thickness in both positions was 2.84 mm (SD 0.41 mm, range 2.0-4.3 mm). Mean thickness in the decubitus position was found to be significantly larger than in the elbow position (P < 0.01). Lateral decubitus expiratory radiography showed physiological pleural fluid in only one case. CONCLUSION: Chest ultrasonography is superior to lateral decubitus expiratory chest radiography for demonstrating small amounts of normal pleural fluid in healthy individuals. A positive result, if detected, should not be taken as a sign of occult thoracic disease. PMID- 15351249 TI - Common crus aplasia: diagnosis by 3D volume rendering imaging using 3DFT-CISS sequence. AB - AIM: The purpose of this study was to evaluate the findings of three-dimensional (3D) volume rendering (VR) imaging in common crus aplasia (CCA) of the inner ear. MATERIALS AND METHODS: Using 3D VR imaging of temporal bone constructive interference in steady state (CISS) magnetic resonance (MR) images, we retrospectively reviewed seven inner ears of six children who were candidates for cochlear implants and who had been diagnosed with CCA. As controls, we used the same method to examine 402 inner ears of 201 patients who had no clinical symptoms or signs of sensorineural hearing loss. Temporal bone MR imaging (MRI) was performed with a 1.5 T MR machine using a CISS sequence, and VR of the inner ear was performed on a work station. Morphological image analysis was performed on rotation views of 3D VR images. RESULTS: In all seven cases, CCA was diagnosed by the absence of the common crus. The remaining superior semicircular canal (SCC) was normal in five and hypoplastic in two inner ears, while the posterior SCC was normal in all seven. One patient showed bilateral symmetrical CCA. Complicated combined anomalies were seen in the cochlea, vestibule and lateral SCC. CONCLUSION: 3D VR imaging findings with MR CISS sequence can directly diagnose CCA. This technique may be useful in delineating detailed anomalies of SCCs. PMID- 15351250 TI - Migrated foreign body granulomas on mammography after injection in the cervicofacial area. AB - AIM: The purpose of this study was to describe the mammographic findings of localized foreign body granulomas in women who were injected with a foreign body in the cervicofacial area to flatten wrinkles. MATERIALS AND METHODS: The teaching files of our institutions were searched, and nine mammograms of migrated foreign body granulomas located. The mammograms were analysed, including the location, shape, size, distribution and depth of the high-density nodules. All these cases had undergone sonography. RESULTS: The migrated foreign body granulomas appeared bilateral (n = 6) or unilateral (n = 3), as oval or round high-density nodules, in the upper inner and/or upper central locations. The average size of the lesions was less than 5 mm in all cases. All nine sonograms showed multiple, round cystic nodules, less than 5 mm in size, with strong posterior shadowing from far upper central to the upper inner breast. Sonography of the lower neck revealed the anatomy could not be visualized due to the strong posterior shadowing in the skin and subcutaneous fat layer. CONCLUSION: Migrated foreign body granulomas appeared on mammograms as localized, multiple oval or round high-density nodules, less than 0.5 cm in size, in the upper inner and/or upper central breast. Breast sonography, including the sonography of the suspected cervicofacial area, is helpful in verifying migrated foreign body granulomas. PMID- 15351251 TI - Managing errors in radiology: a working model. AB - AIM: To develop a practical mechanism for reviewing reporting discrepancies as addressed in the Royal College of Radiologists publication "To err is human. The case for review of reporting discrepancies". MATERIALS AND METHODS: A regular meeting was developed, and has evolved, within the department to review discrepancies. Standard forms were devised for submission of cases as well as recording and classification of discrepancies. This has resulted in availability of figures that can be audited annually. RESULTS: Eighty-one cases involving error were reviewed over a 12-month period. Seven further cases flagged as discrepancies were not identified on peer review. Twenty-four reports were amended subsequent to the meeting. Nineteen additional cases were brought to the meeting as illustrative of teaching points or for discussion. CONCLUSION: We have evolved a successful process of reviewing reporting errors, which enjoys the confidence and support of all clinical radiologists, and is perceived as a method of improving patient care through an increasing awareness of lapses in performance. PMID- 15351252 TI - Diagnostic value of MRCP in the management of hilar strictures after extended liver resection. PMID- 15351253 TI - Malignant monotypic epithelioid angiomyolipoma of the kidney. PMID- 15351254 TI - Conscious sedation for endoscopic and non-endoscopic interventional procedures: meeting patients' expectations, missing the standard. PMID- 15351256 TI - Mammographic casting-type calcification associated with small screen-detected invasive breast cancers: is this a reliable prognostic indicator? PMID- 15351257 TI - To clot or not to clot? That is the question in central venous catheters. PMID- 15351258 TI - To clot or not to clot? That is the question in central venous catheters. PMID- 15351260 TI - To clot or not to clot? That is the question in central venous catheters. PMID- 15351261 TI - How to synthesize evidence for imaging guidelines. PMID- 15351263 TI - How to synthesize evidence for imaging guidelines. PMID- 15351266 TI - Destabilization of phospholipid model membranes by YplA, a phospholipase A2 secreted by Yersinia enterocolitica. AB - Yersinia enterocolitica produces a virulence-associated phospholipase A(2) (YplA) that is secreted via its flagellar type-III secretion apparatus. When the N terminal 59 amino acids of YplA are removed (giving YplA(S)), it retains phospholipase activity; however, it is altered with respect to the apparent kinetics of hydrolysis using fluorescent phospholipid substrates in micellar form. To explore the physical properties of YplA more carefully, Langmuir phospholipid monolayers were used to study the association of YplA with biological membranes. YPlA and YplA(S) both associate with Langmuir monolayers, but YplA(S) appears to interact better at low initial lipid densities while YplA interacts better at higher densities. This may indicate that the N-terminus of YplA has a role in mediating its initial interaction with compact cellular membranes, which is consistent with spectroscopic observations that fluorescein labeled YplA may interact more readily with the nonpolar region of liposomes than does YplA(S). PMID- 15351267 TI - Interaction of 3,7-diamino-2,8-dimethyl-5-phenyl phenazinium chloride with model biological membranes and reverse micelles of lipid: a spectroscopic study. AB - The interaction of 3,7-diamino-2,8-dimethyl-5-phenyl phenazinium chloride (Safranine T) with the aqueous as well as reverse micellar solution of a phospholipid 1,2-diacyl-sn-glycero-3-phosphocholine (Azolecithin), a major structural phospholipid in brain, comprising approx 15% of total lipid, primarily localized in grey matter have been studied by absorption and fluorescence spectroscopic studies. The results show the evidence of complex formation of the dye in the ground and in the excited state. The interaction of the dye with the lipid in reverse micellar state is more compared to that in liposomes. An attempt has been made to determine the polarity of the microenvironment of the dye in liposomes or reverse micelles from the spectral studies of the dye in different solvents of known polarity. The polarity functions of the phosphatidylcholine (PC) liposomes are slightly lower compared to that of PC reverse micelles. PMID- 15351268 TI - Short synthesis of labeled and unlabeled 6Z,9Z,12Z,15-hexadecatetraenoic acid as metabolic probes for biosynthetic studies on diatoms. AB - We describe a short synthesis of the unusual polyunsaturated 6Z,9Z,12Z,15 hexadecatetraenoic acid found in marine and fresh water diatoms. Using a one pot reductive bis-Wittig-olefination, the homoconjugated tetraene backbone of the fatty acid can be generated from easy available precursors. Reductive olefination allows the non-statistical dissymmetrisation of a symmetrical bis-Wittig salt as key synthon. This short sequence was also applied to the generation of the corresponding 9,10-[(2)H(2)] labeled fatty acid. If administered to cell fragments of Thalassiosira rotula 9,10-[(2)H(2)]-6Z,9Z,12Z,15-hexadecatetraenoic acid is transformed oxidatively to the aldehyde 1,2-[(2)H(2)]-2E,4E/Z,7 octatrienal which is involved in the chemical defense of this alga. Using the synthetic standard it could be shown that the C16:4 omega1 fatty acid is released upon wounding of T. rotula cells. The synthesis with the labeled bis-Wittig salt is of general use and can also be applied for the fast generation of other internally labeled functionalized and non-functionalized polyunsaturated fatty acids. To our knowledge this represents the first synthesis of 6Z,9Z,12Z,15 hexadecatetraenoic acid. PMID- 15351269 TI - Temperature and cholesterol composition-dependent behavior of 1-myristoyl-2-[12 [(5-dimethylamino-1-naphthalenesulfonyl)amino]dodecanoyl]-sn-glycero-3 phosphocholine in 1,2-dimyristoyl-sn-glycero-3-phosphocholine membranes. AB - We present a steady-state and time-resolved fluorescence emission spectra analysis of the membrane probe 1-myristoyl-2-[12-[(5-dimethylamino-1 naphthalenesulfonyl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (DANSYL) in 1,2 dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and cholesterol multi-lamellar vesicles (MLV) prepared by modified rapid solvent exchange. We report that the dose-dependent cholesterol-induced blue shifts in the steady-state fluorescence emission spectra observed in DMPC MLV are due to complex solvent effects that include time-dependent dipolar relaxation and the formation of internal charge transfer (ICT) states. A key finding of this investigation is identification of two distinguishable DANSYL populations existing at both shallow and deep locations in the membrane; these two DANSYL populations are evidence of laterally phase-separated domains at cholesterol compositions between X(chol) = 0.30 and 0.60 at 30 degrees C in DMPC MLV. PMID- 15351270 TI - The interaction of water with the phospholipid head group and its relationship to the lipid electrical conductivity. AB - We have studied the interaction of water with the lipid head group by gravimetrically measuring the lipid water adsorption and the lateral dc electrical conductivity increase resulting from this hydration. We have done this for dimyristoyl phosphatidylcholine (DMPC) having protonated or deuterated hydrocarbon chains. These studies were also done for two cationic lipids having rather different polar head groups. All three lipids behave as strong water adsorbers and all three display a steep, logarithmic increase in the conductivity as the first 1-3 waters per lipid are adsorbed. This increase is usually 5-6 orders of magnitude. After the initial 1-3 waters are adsorbed, the conductivity increases much more gradually, upon additional water adsorption. This electrical behavior is also found for weak water adsorbers and appears to be independent of the head group composition. The conductivity behavior suggests two types of water interacting with the head group. Our studies also indicate that a choline-like component is responsible for the strong water binding nature of the lipids, although, both phosphate and choline make significant contributions to the total amount of adsorbed water. The conductivity behavior, however, does not depend on the presence of both these head group components. PMID- 15351271 TI - Structural characterization of diC14-amidine, a pH-sensitive cationic lipid used for transfection. AB - The structure of N-t-butyl-N'-tetradecyl-3-tetradecylaminopropionamidine (diC(14) amidine) cationic vesicles, used for transfection, was investigated at different pH values and ionic strengths, through the analysis of the electron spin resonance (ESR) spectra of spin labels. Phospholipid derivatives, spin labeled at the 5th and 16th C-atoms along the hydrocarbon chain, incorporated in diC(14) amidine bilayers, show that the bilayer structure is highly sensitive to the pH value of the medium, due to the two titratable groups present in the amphiphile. Compared with samples at higher pH values, the double charged diC(14)-amidine at pH 3 presents a rather non-organized bilayer gel phase, and a much lower gel fluid temperature transition, in accord with a strong headgroup electrostatic repulsion. In addition, the structure was found to be highly dependent on the ionic strength of the medium. However, pH 3 diC(14)-amidine bilayer, in the fluid phase, was found to be slightly more closely packed than those at pH 7.4 or 9.0, which are less charged. Parallel to that, the larger isotropic hyperfine splitting measured for nitroxides in the center of the pH 3 diC(14)-amidine bilayer suggests a higher membrane polarity for the highly charged low pH sample. PMID- 15351272 TI - Molecular species of ceramides from the ascomycete truffle Tuber indicum. AB - The ceramide fractions were isolated from the chloroform/methanolic extractable of the fruiting bodies of Tuber indicum and separated into three kinds of molecular species TI-1, TI-2, and TI-3 by normal and reverse phase silica gel column chromatography. By means of (1)H NMR and (13)C NMR spectroscopy, fast atom bombardment mass spectrometry (FAB-MS), and chemical degradation experiment, their component sphingoid base for TI-1 and TI-2 was uniformly (2S,3S,4R)-2-amino 1,3,4-octadecantriol, while the sphingoid of TI-3 was d-erythro-sphingosine, and their structures have been determined unequivocally to be (2S,2'R,3S,4R)-2-(2'-d hydroxyalkanoylamino) octadecane-1,3,4-triol, the fatty acid composition of which consists of 2-hydroxydocosanoic, 2-hydroxytetracosanoic, and 2-hydroxytricosanoic acids (from major to minor); (2S,3S,4R)-2-(alkanoylamino)octadecane-1,3,4-triol, the fatty acid composition of which is unusual and consists of docosanoic, hexadecanoic, tricosanoic, octadecanoic and nonadecanoic acids (from major to minor); and (2S,3R,4E)-2-(alkanoylamino)-4-octadecene-1,3-diol, the component fatty acids of which were hexadecanoic (predominant) and octadecanoic acids, respectively. PMID- 15351273 TI - Synthesis of four isomers of parinaric acid. AB - A simple and reliable method for synthesizing four isomers of parinaric acid from alpha-linolenic acid (ALA) in high yields is described. The methylene interrupted, cis triene system (1,4,7-octatriene) of ALA and common to other naturally occurring polyunsaturated fatty acids was transformed to a conjugated tetraene system (1,3,5,7-octatetraene). The synthesis involves bromination of ALA using 0.l M Br(2) in a saturated solution of NaBr in methanol, esterification of the fatty acid dibromides, double dehydrobromination by 1,8 diazabicyclo[5.4.0]undec-7-ene and saponification of the conjugated esters to a mixture of free conjugated acids. Addition of one molecule of bromine to the 12,13-double bond of ALA and subsequent dehydrobromination produces alpha parinaric acid (9Z,11E,13E,15Z-octadecatetraenoic acid); addition of Br(2) to the 9,10-double bond or 15,16-double bond and then dehydrobromination and rearrangement yields 9E,11E,13E,15Z-octadecatetraenoic or 9E,11E,13E,15Z octadecatetraenoic acids, respectively. The mixture of parinaric acid isomers is obtained in 65% yield, and the isomers can be purified by preparative HPLC; alternatively, the isomers can be converted by base catalyzed cis-trans isomerization (or by treatment with I(2)) to exclusively beta-parinaric acid (9E,11E,13E,15E-octadecatetraenoic acid). The various parinaric acid isomers were characterized by (1)H NMR, (13)C NMR, UV, GLC, HPLC and mass spectrometry. PMID- 15351274 TI - History and principles of conductive media for standard DNA electrophoresis. AB - DNA electrophoresis has been a dominant technique in molecular biology for 30 years. The foundation for this common technique is based on a few simple electrochemical principles. Electrophoretic DNA separation borrowed from existing protein and RNA techniques developed in the 1950s and 1960s. For 30 years, common DNA electrophoretic conductive media remained largely unchanged, with Tris as the primary cation. DNA electrophoresis relies simply upon the negative charge of the phosphate backbone and the ability to distribute a voltage gradient in a sieving matrix. Nevertheless, the conductive properties in DNA electrophoresis are complicated by choices involving voltage, electric current, conductivity, temperature, and the concentration and identity of the ionic species present. Differences among the extant chemical recipes for common conductive media affect central properties. Tris-based buffers, even in optimal form, create a runaway positive feedback loop between heat generation and retention, temperature, conductivity, and current. This is undesirable, leading to limitations on the permissible electric field and to impaired resolution. Recently, we developed low molarity conductive media to mitigate this positive feedback loop. Such media allow for application of a higher electric field. Applications of DNA electrophoresis can now be reengineered for lower ionic strength, higher field strengths, and lower requirements for heat dissipation. PMID- 15351275 TI - A coupled spectrophotometric enzyme assay for the determination of pectin methylesterase activity and its inhibition by proteinaceous inhibitors. AB - Pectin methylesterase (PME; EC 3.1.1.11) activities are widespread in bacteria, fungi, and plants. PME-mediated changes in cell wall pectin structure play important roles in plant development. Genome sequencing projects have revealed the existence of large PME multigene families in higher plants. Additional complexity for PME regulation arises from the presence of specific PME inhibitor proteins (PMEI) in plant cells. Several assay procedures for the determination of PME activity have been reported. However, previous protocols suffered from various limitations. Here we report a protocol for a coupled enzyme assay based on methanol oxidation via alcohol oxidase (AO; EC 1.1.3.13) and subsequent oxidation of formaldehyde by formaldehyde dehydrogenase (FDH; EC 1.2.1.3). This simple and robust assay allows the continuous monitoring of PME activity in the neutral pH range. Furthermore, as plant PMEIs do not interfer with AO and FDH activities, this assay is suitable for the characterization of the inhibition kinetics of PMEI. PMID- 15351276 TI - A differential assay for the reduced and oxidized states of metallothionein and thionein. AB - In the cellular environment, the sulfur ligands in zinc/thiolate coordination sites of proteins can be oxidized with concomitant mobilization of zinc. The characterization of such "redox zinc switches" requires the determination of three species, i.e., the zinc-containing complex and the zinc-free complex with the thiolate ligands either reduced or oxidized. Differential chemical modification of thiol groups in the presence and absence of either reducing or chelating agents allows the analytical speciation of such systems as demonstrated here for the characterization of the redox and metal-binding states of mammalian metallothionein. Thiol derivatization with 6-iodoacetamidofluorescein in the presence and absence of the reducing agent tris(2-carboxyethyl)phosphine, high performance liquid chromatographic separation, and photometric detection are employed to determine the reduced and oxidized protein. Because the holoprotein reacts only in the presence of a chelating agent such as ethylenediaminetetraacetate (EDTA) its amount can be determined as the difference between measurements in the presence and the absence of EDTA. This method is applied to the study of the chemical and enzymatic oxidation of metallothionein/thionein. It should also greatly facilitate the characterization of the redox and metal-binding properties of zinc/thiolate coordination environments of other proteins such as zinc finger proteins. PMID- 15351277 TI - High-performance liquid chromatography-based methods of enzymatic analysis: electron transport chain activity in mitochondria from human skeletal muscle. AB - This study addresses an application of pyridine nucleotide enzymatic analyses to evaluate the activity of the mitochondrial electron transport chain (reduced nicotinamide adenine dinucleotide (NADH) oxidase) and Complexes I and II in samples of human muscle as small as approximately 10 mg wet weight. Key aspects in this adaptation are the use of high-performance liquid chromatography with fluorescence detection of NADH and use of alamethicin, a channel-forming antibiotic that enables an unrestricted access of substrates into the mitochondrial matrix. The procedure includes disintegration of tissue by Polytron homogenizer, extraction of myosin from myofibrillar fragments by KCl/pyrophosphate to facilitate release of mitochondria, and preparation of fractions of subsarcolemmal and intermyofibrillar mitochondria. Oxidation of NADH or succinate is assayed in the presence of 40 microg/ml alamethicin and the reaction is terminated by H(2)SO(4), which also destroys the remaining NADH. Nicotinamide adenine dinucleotide (NAD) or fumarate concentrations are measured using alcohol dehydrogenase or fumarase plus malic dehydrogenase reactions, respectively. Generation of NADH, assessed in auxiliary reactions in the presence of hydrazine, is strictly proportional to NAD or fumarate content across a concentration range of 1-20 microM. NADH is quantitatively analyzed with a detection limit of 3-5 pmol by HPLC using a reverse-phase Hypersil ODS column connected to a fluorescence detector. PMID- 15351278 TI - Development and validation of a time-resolved fluorometric immunoassay for screening of antichlamydial activity using a genus-specific europium-conjugated antibody. AB - The lack of high-throughput assays has limited the screening of new antimicrobials against obligate intracellular bacteria, including chlamydia, which cause a variety of diseases. In this study, a novel technological approach was developed to detect intracellular bacteria using time-resolved fluorometric immunoassay (TR-FIA), and the method was validated for susceptibility testing of Chlamydia pneumoniae. In this cell-based, 96-well plate assay, chlamydial inclusions are labeled with europium-conjugated antibody and quantified as time resolved fluorometric signals by means of a multilabel counter. To confirm the reliability of the TR-FIA, susceptibilities of C. pneumoniae reference strain Kajaani 7 to a set of antimicrobial agents were determined by the TR-FIA, conventional immunofluorescence staining, and real-time polymerase chain reaction. Minimum inhibitory concentrations measured using the different methods demonstrated good to excellent correlation. Data relating to reproducibility (day to-day variation 9.0%), as well as to the signal-to-background, signal-to-noise, and Z' values (6.5, 6.9, and 0.4, respectively), showed the suitability of the TR FIA for screening. By means of dual labeling with sulfornodamine B the cytotoxicity of test compounds could be detected simultaneously with the susceptibility testing. In summary, the TR-FIA is a convenient, reliable, and objective alternative for detecting chlamydia in vitro. By being considerably less labor intensive and offering significantly higher throughput, the TR-FIA is especially suitable for screening of new antichlamydial compounds. PMID- 15351279 TI - Glucose biosensor prepared by glucose oxidase encapsulated sol-gel and carbon nanotube-modified basal plane pyrolytic graphite electrode. AB - A new glucose biosensor has been fabricated by immobilizing glucose oxidase into a sol-gel composite at the surface of a basal plane pyrolytic graphite (bppg) electrode modified with multiwall carbon nanotube. First, the bppg electrode is subjected to abrasive immobilization of carbon nanotubes by gently rubbing the electrode surface on a filter paper supporting the carbon nanotubes. Second, the electrode surface is covered with a thin film of a sol-gel composite containing encapsulated glucose oxidase. The carbon nanotubes offer excellent electrocatalytic activity toward reduction and oxidation of hydrogen peroxide liberated in the enzymatic reaction between glucose oxidase and glucose, enabling sensitive determination of glucose. The amperometric detection of glucose is carried out at 0.3 V (vs saturated calomel electrode) in 0.05 M phosphate buffer solution (pH 7.4) with linear response range of 0.2-20 mM glucose, sensitivity of 196 nA/mM, and detection limit of 50 microM (S/N=3). The response time of the electrode is < 5s when it is stored dried at 4 degrees C, the sensor showed almost no change in the analytical performance after operation for 3 weeks. The present carbon nanotube sol-gel biocomposite glucose oxidase sensor showed excellent properties for the sensitive determination of glucose with good reproducibility, remarkable stability, and rapid response and in comparison to bulk modified composite biosensors the amounts of enzyme and carbon nanotube needed for electrode fabrication are dramatically decreased. PMID- 15351280 TI - Purification of RanGDP, RanGTP, and RanGMPPNP by ion exchange chromatography. AB - Ran is a small GTPase that cycles between a guanosine diphosphate (GDP)-bound form (RanGDP) and a guanosine triphosphate (GTP)-bound form (RanGTP) and plays important roles in nuclear transport and mitosis. For studies of Ran function and its interactions with partner proteins, pure RanGDP and RanGTP complexes are critical. Ran complexed with the nonhydrolyzable GTP analog, GMPPNP (RanGMPPNP), is used instead of RanGTP when inhibition of hydrolysis is required. In this study, we demonstrate that the binding of Ran to a UNO Q ion exchange column is remarkably sensitive to small shifts in MgCl(2) concentration, and we use this property to purify recombinant RanGTP, RanGMPPNP, and RanGDP complexes. At 10 mM MgCl(2), Ran was found predominantly in the flow-through and, thus, was separated from the vast majority of bacterial proteins. After reducing the concentration of MgCl(2) to 5 mM, further purification of RanGTP, RanGMPPNP, and RanGDP was achieved by loading onto ion exchange columns and elution with an NaCl gradient. Purity of the resulting preparations was confirmed by releasing the bound nucleotide and checking it against a known nucleotide by high-performance liquid chromatography (HPLC). To further confirm the purity and function of the Ran preparations, appropriate protein-binding, enzymatic, and nuclear import assays were carried out. These methods should facilitate studies of cellular processes involving Ran by providing pure functional Ran-nucleotide complexes. PMID- 15351281 TI - Measurement of DNA adducts in cells exposed to cisplatin. AB - The two main cisplatin-induced DNA lesions, G--G and A--G, have been measured in cells exposed to the drug. (G--G and A--G denote the intrastrand bifunctional adducts formed between adjacent purine bases.) It has proven feasible, using liquid chromatography-tandem mass spectrometry (LC-MS/MS), observe the G--G and A -G lesions in mouse fibroblast cells exposed for 1 h to a 120 microM concentration of cisplatin. After extraction of the DNA from the cells, the lesions were enzymatically isolated from the DNA in the form of modified dinucleoside monophosphates with the phosphodiester bond intact. MS/MS detection of the modified dinucleoside monophosphates in the negative ion mode manifests two transitions; from the negative ion to the loss of one NH(3) group and from the ion less one NH(3) group to the loss of both NH(3) groups. The multiple reaction monitoring capability of LC-MS/MS was used to measure the three most abundant isotopes of the two main lesions for both transitions of each lesion (i.e., 12 MS/MS values in toto). Ion currents could be detected for all 12 pairs of MS/MS values in the DNA from exposed cells. Although this protocol results in some overlap of MS/MS values between the two lesions, a slight difference in elution times clearly distinguishes between them. PMID- 15351282 TI - Estimation of relative allele frequencies of single-nucleotide polymorphisms in different populations by microarray hybridization of pooled DNA. AB - Single-nucleotide polymorphisms (SNPs) are considered useful polymorphic markers for genetic studies of polygenic traits. A new practical approach to high throughput genotyping of SNPs in a large number of individuals is needed in association study and other studies on relationships between genes and diseases. We have developed an accurate and high-throughput method for determining the allele frequencies by pooling the DNA samples and applying a DNA microarray hybridization analysis. In this method, the combination of the microarray, DNA pooling, probe pair hybridization, and fluorescent ratio analysis solves the dual problems of parallel multiple sample analysis, and parallel multiplex SNP genotyping for association study. Multiple DNA samples are immobilized on a slide and a single hybridization is performed with a pool of allele-specific oligonucleotide probes. The results of this study show that hybridization of microarray from pooled DNA samples can accurately obtain estimates of absolute allele frequencies in a sample pool. This method can also be used to identify differences in allele frequencies in distinct populations. It is amenable to automation and is suitable for immediate utilization for high-throughput genotyping of SNP. PMID- 15351283 TI - Liquid chromatographic and tandem mass spectrometric assay for evaluation of in vivo inhibition of rat brain monoamine oxidases (MAO) A and B following a single dose of MAO inhibitors: application of biomarkers in drug discovery. AB - A simple and selective assay for the evaluation of in vivo inhibition of rat brain monoamine oxidases (MAO) A and B following a single dose of MAO inhibitors was developed through the simultaneous determination of endogenous 5-hydroxy tryptamine, 5-hydroxyindole-3-acetic acid (5-HIAA), tryptophane, and 2 phenethylamine (PEA) in rat brain using liquid chromatography-tandem mass spectrometry (LC/MS/MS). These analytes were separated on a Zorbax SB-C18 column using a gradient elution with acetonitrile and 0.2% formic acid and detected on an electrospray ionization mass spectrometer in positive-ion multiple-reaction monitoring mode. The susceptibility and variability of these analytes as potential biomarkers in response to MAO inhibition in vivo were evaluated after application to three MAO inhibitors, tranylcypromine, clorgyline, and pargyline. A dramatic increase (about 40-fold) in PEA brain level and a decrease in 5-HIAA by more than 90% were observed after administration of 15 mg/kg of the nonselective MAO inhibitor tranylcypromine. As expected, the brain level of PEA escalated to about 6-fold, while the 5-HIAA level remained unchanged following a dose of the MAO B inhibitor pargyline at 2mg/kg. In contrast, the brain level of 5-HIAA reduced by approximately 53%, but the PEA level was unaffected following the same dose of the MAO A inhibitor clorgyline. The results indicated that 5 HIAA and PEA were susceptible and effective biomarkers in the rat brain in response to MAO A and B inhibition, respectively. The LC/MS/MS method is useful not only for the determination of inhibitory potency but also for the differentiation of the selectivity of a MAO inhibitor against rat brain MAO A and B in vivo. PMID- 15351284 TI - Amperometric cholesterol biosensor based on in situ reconstituted cholesterol oxidase on an immobilized monolayer of flavin adenine dinucleotide cofactor. AB - A new amperometric biosensor for determining cholesterol based on deflavination of the enzyme cholesterol oxidase (ChOx) and subsequent reconstitution of the apo protein with a complexed flavin adenine dinucleotide (FAD) monolayer is described. The charge transfer mediator pyrroquinoline quinone (PQQ) was covalently bound to a cystamine self-assembled monolayer (SAM) on an Au electrode. Boronic acid (BA) was then bound to PQQ using the carbodiimide procedure, and the BA ligand was complexed to the FAD molecules on which the apo ChOx was subsequently reconstituted. The effective release of the FAD from the enzyme and the successful reconstitution were verified using molecular fluorescence and cyclic voltammetry. The optimal orientation of FAD toward the PQQ mediator and the distances between FAD and PQQ and between PQQ and electrode enhance the charge transfer, very high sensitivity (about 2,500 nAmM(-1)cm(-2)) being obtained for cholesterol determination. The biosensor is selective toward electroactive interferents (ascorbic acid and uric acid) and was tested in reference serum samples, demonstrating excellent accuracy (relative errors below 3% in all cases). The biosensor activity can be successfully regenerated in a simple process by successive reconstitution with batches of recently prepared apo ChOx on the same immobilized Au/SAM-PQQ-BA-FAD monolayer (it was tested five times); the lifetime of the biosensor is about 45-60 days. PMID- 15351285 TI - Sensitivity-enhancement of wavelength-modulation surface plasmon resonance biosensor for human complement factor 4. AB - Sandwich and colloidal Au techniques for enhancing the sensitivity of a wavelength-modulation surface plasmon resonance (SPR) immunosensor are demonstrated by the detection of human complement factor 4 (C4). The design of the wavelength-modulation SPR biosensor is based on fixing the incident angle of light and measuring the reflected intensity of light in the wavelength range spanning 500-900 nm simultaneously. The human C4 had good response in the concentration range 2-20 microg/mL in the direct assay. However, in the sandwich assay, the human C4 had good response in the concentration range 0.2-20 microg/mL and the lowest concentration is 10-fold lower than that obtained by the direct assay. With human C4-Au colloidal conjugate, the human C4 had good response in the concentration range 0.1-20 microg/mL and the lowest concentration is 20-fold lower than that obtained by the direct assay. In the colloidal-Au-enhanced sandwich assay, the human C4 had good response in the concentration range 0.05-5 microg/mL and the lowest concentration is 40-fold lower than that obtained by the direct assay. Under selected experimental conditions, the reproducibility, sensitivity, and reversibility of the enhanced SPR immunoassay are very satisfactory. The results represent potentially significant advantages in the sensitivity of SPR biosensors. PMID- 15351286 TI - A homogeneous time-resolved fluorescence detection of telomerase activity. AB - The homogeneous time-resolved fluorescence (HTRF) technology is an assay developed to study the interaction between biomolecules. This detection system is based on a fluorescence resonance energy transfer (FRET) between a Tris bipyridine europium cryptate used as a long-lived fluorescent donor and a chemically modified allophycocyanine as acceptor. This technology is characterized by both a spectral selectivity and a temporal selectivity (due to the time-resolved mode), ensuring a highly specific signal. Here a europium cryptate-labeled deoxyuridine triphosphate analogue (K-11-dUTP) was used to monitor the extension reaction on a biotinylated oligonucleotide used as substrate for telomerase in a telomeric repeat amplification protocol (TRAP). After the addition of an allophycocyanine-streptavidin conjugate, the extension products give rise to a FRET between the incorporated cryptate moieties and the allophycocyanine acceptor that then displays a specific long-lived emission. The TRAP-HTRF format was validated as a screening tool by using a 2,6 diaminoanthraquinone analogue, a known inhibitor of telomerase activity. The IC(50) measured was consistent with the reported values, showing the convenience of the HTRF technology for the study of telomerase activity and inhibitors. PMID- 15351287 TI - Limitations of the ultracentrifugal approach for measuring the effective net charge of a macroion. AB - Limitations have been detected in a recently published method for macroion valence determination by an ultracentrifugal procedure for quantifying the Donnan distribution of small ions in macroion solutions dialyzed against buffer supplemented with chromate as an indicator ion. The limitation reflects an implicit assumption that sedimentation velocity affords an unequivocal means of separating effects of chromate binding from those reflecting the Donnan redistribution of small ions. Although the assumed absence of significant Donnan redistribution of small ions across the sedimenting macroion boundary seemingly holds for some systems, this approximation is demonstrably invalid for others. Despite preliminary signs of promise, the ultracentrifugal procedure does not afford a simple, readily applied solution to the problem of unequivocal macroion valence determination. PMID- 15351288 TI - De novo quantitative bisulfite sequencing using the pyrosequencing technology. AB - Current protocols for DNA methylation analysis are either labor intensive or limited to the measurement of only one or two CpG positions. Pyrosequencing is a real-time sequencing technology that can overcome these limitations and be used as an epigenotype-mapping tool. Initial experiments demonstrated reliable quantification of the degree of DNA methylation when 2-6 CpGs were analyzed. We sought to improve the sequencing protocol so as to analyze as many CpGs as possible in a single sequencing run. By using an improved enzyme mix and adding single-stranded DNA-binding protein to the reaction, we obtained reproducible results for as many as 10 successive CpGs in a single sequencing reaction spanning up to 75 nucleotides. A minimum amount of 10 ng of bisulfite-treated DNA is necessary to obtain good reproducibility and avoid preferential amplification. We applied the assay to the analysis of DNA methylation patterns in four CpG islands in the vicinity of IGF2 and H19 genes. This allowed accurate and quantitative de novo sequencing of the methylation state of each CpG, showing reproducible variations of methylation state in contiguous CpGs, and proved to be a useful adjunct to current technologies. PMID- 15351289 TI - Biosynthesis of [3H]7 alpha-hydroxy-, 7 beta-hydroxy-, and 7-oxo dehydroepiandrosterone using pig liver microsomal fractions. AB - Current research on dehydroepiandrosterone (DHEA) is limited due to lack of radiolabeled metabolites. We utilized pig liver microsomal (PLM) fractions to prepare [(3)H]-labeled 7 alpha-hydroxy-DHEA (7 alpha-OH-DHEA), 7 beta-hydroxy DHEA (7 beta-OH-DHEA), and 7-oxo-DHEA substrates from 50 microM [1,2,6,7 (3)H]DHEA (specific radioactivity 60-80 mCi/mmol). The metabolites were separated by preparative thin-layer chromatography (TLC) using ethyl acetate:hexane:glacial acetic acid (18:8:3 v:v:v) as the mobile phase, extracted with ethyl acetate, and dried under a stream of nitrogen. Metabolites assayed by TLC and gas chromatography-mass spectrometry were observed to be pure. In the presence of an reduced nicotinamide adenine dinucleotide phosphate (NADPH)-regenerating system initiated with 1 mM NADPH alone, 1 mg/ml PLM produced 7 alpha-OH-DHEA with minor amounts of 7-oxo-DHEA (68 and 14 nmol/2h/2 ml, respectively; 82% conversion), while in the presence of 1mM NADPH and 1 mM oxidized nicotinamide adenine dinucleotide phosphate (NADP(+)), more 7-oxo-DHEA than 7 alpha-OH-DHEA (58 and 11 nmol/2 ml/120 min, respectively; 69% conversion) was formed. When longer reaction times were used with NADPH and NADP(+), a mixture of 7 alpha-OH-DHEA, 7 beta-OH DHEA, and 7-oxo-DHEA was produced (19,14, and 35 nmol/180 min/2 ml, respectively; 62% conversion). Using pig liver microsomes, the radiolabeled metabolites of DHEA can be prepared in stable, pure form at 10mM concentrations and >0.5 mCi/mmol levels of radioactivity for biochemical studies. PMID- 15351290 TI - Modeling of homogeneous cloned enzyme donor immunoassay. AB - One of the most widely used analytical techniques for sensitive detection of biologically and clinically significant analytes is the immunoassay. In recent years direct immunoprobes allowing label-free detection of the interaction between the antibody and the target analyte have proved their capabilities as fast, simple, and nevertheless highly sensitive methods. Cloned enzyme donor immunoassay (CEDIA) homogeneous assay is based on the bacterial enzyme beta galactosidase, which has been genetically engineered into two inactive fragments, enzyme donor and enzyme acceptor. Reassociation of the fragments in the assay forms active enzyme, which acts on substrate to generate a colored product. A comprehensive kinetic model of CEDIA is developed to aid in understanding this method and to facilitate development of a truly homogeneous version, potentially applicable to a dipstick-type multianalyte point of care analytical device (ChemChip). Although the standard assay involves a two-step process, we also chose to model a single-combined process, which would be simpler to apply in a ChemChip device. From the modeling simulation, we obtain the time courses of the amounts of product and active enzyme, from which the dynamic ranges can be obtained as 10(-6)-10(-7) and 10(-5)-10(-7)M analyte concentration for two-step and single-combined processes under the conditions of the assumed parameters, respectively. A simple one-step immunoassay has the merit of reducing time and cost and has an improved dynamic range. PMID- 15351291 TI - Fluorometric polyethyleneglycol-peptide hybrid substrates for quantitative assay of protein disulfide isomerase. AB - In eukaryotic cells the enzyme protein disulfide isomerase (PDI) is responsible for the formation and reshuffling of disulfide bonds in secretory proteins. The reaction carried out by PDI involves interaction with a highly complex mixture of polypeptide molecules that are in the process of folding. This means that PDI activity is typically measured in the context of a globular protein folding pathway. The absence of small, well-defined substrates for the quantitation of both oxidation and reduction reactions constitutes an inherent problem in the analysis of PDI activity. We describe a new type of substrate for PDI where two cysteine-containing oligopeptides are connected by an onameric ethylene glycol linker. We term such hybrid compounds PEGtides. The oligopeptides are each marked with a fluorescent aminobenzoic acid and a quenching nitrotyrosine group, respectively. The reversible formation of an intramolecular disulfide bond between fluorophore-containing and quencher-containing peptide segments results in a redox-dependent fluorescence signal. We find a model compound of this type to be a highly sensitive substrate for PDI both in oxidation and in reduction assays under steady state conditions. These aspects should make substrates of this type generally applicable for assaying PDI and other thiol-disulfide exchange enzymes. PMID- 15351292 TI - Mass spectrometric quantitation of deoxyguanosine and leukotriene A4 deoxyguanosine adducts of DNA. AB - An assay was developed using electrospray ionization negative ion tandem mass spectrometry (MS) to identify and quantitate the major product in the reaction of leukotriene A(4) (LTA(4)) with deoxyguanosine (dGuo). A second quantitative assay was established using the same separation and detection techniques to determine the amount of dGuo isolated from enzymatically processed DNA. The amount of LTA(4)-dGuo adduct could then be analytically determined in DNA samples and normalized to the amount of dGuo that had been simultaneously derived from the DNA sample. Stable isotope-labeled internal standards used for these quantitative assays were readily synthesized from isotopically labeled [(15)N(5)(13)C(10)]deoxyguanosine triphosphate and analyzed for isotopic purity using MS. A comparison of fragment ions formed from stable isotope analogs of dGuo revealed the loss of deoxyribose and secondarily the loss of a series of stable neutral small molecules in a fashion similar to patterns described previously for the collisional fragmentation of protonated guanine determined by positive ion fast atom bombardment/MS/MS. The combined quantitative assays were used for the determination of the amount of endogenously formed LTA(4)-dGuo adducts observed in DNA when isolated human neutrophils that had been incubated with arachidonic acid were stimulated with calcium ionophore to initiate leukotriene biosynthesis. PMID- 15351293 TI - Signal amplification on planar and gel-type sensor surfaces in surface plasmon resonance-based detection of prostate-specific antigen. AB - This article describes surface plasmon resonance (SPR)-based detection of prostate-specific antigen (PSA), comparing amplification with colloidal gold (10nm diameter) and latex microspheres (120 nm diameter) on planar- and gel-type sensor surfaces. As matrix, 3% BSA in PBS was used. Experimental data were compared with model calculations that predict the SPR signal that results from covering of the different sensor surfaces with each of the particles used. Amplification with latex particles gave a higher signal than did that with colloidal gold. However, the limit of detection (LOD) attained by latex amplification was not as good as that obtained after gold amplification, and this was unexpected. LOD and sensitivity of the amplified PSA assays when performed with the planar-type sensor disc were equally good or better compared with those when performed with the gel-type sensor disc. Indirect evidence indicates a restricted accessibility of the gel layer on the gel-type sensor toward the colloidal gold. Application of colloidal gold led to a sensitivity increase of approximately three orders of magnitude compared with nonamplified detection. The corresponding LOD was approximately 0.15 ng PSA/ml, which is sufficient for measuring enhanced, clinically relevant PSA levels (>4 ng/ml). PMID- 15351294 TI - A simplified procedure for the reduction and alkylation of cysteine residues in proteins prior to proteolytic digestion and mass spectral analysis. AB - A procedure for reduction and alkylation of cysteine residues in proteins was developed using the volatile reagents triethylphosphine and iodoethanol. These reagents may be used to modify proteins in solution, as well as proteins in gel slices, prior to proteolytic digestion and mass spectral analysis. The procedure eliminates several steps with both types of samples. Samples in solution do not need to be desalted following reduction and alkylation, with excess reagent being removed under vacuum. For gel slices, the procedure combines washing, destaining, reduction and alkylation into a single step. The procedure was applied successfully to samples as complex as serum, and we demonstrated alkylation of cysteines to be quantitative in purified proteins. We also were able to reduce and alkylate proteins with these reagents during the gas phase. Elimination of the need for desalting of samples after reaction raised the possibility of automation of the procedure for liquid samples, which is difficult with conventional reduction and alkylation chemistries. PMID- 15351295 TI - An oligonucleotide microarray for the monitoring of repair enzyme activity toward different DNA base damage. AB - Characterization of DNA-N-glycosylase activities in cell extract is a challenging problem and could represent a major concern for medical applications. Synthetic oligonucleotides which contain base lesions located on specific sites constitute suitable substrates for their study. An in vitro miniaturized assay was developed that allows the measurement of cleavage activities of DNA repair enzymes on a set of oligonucleotides (ODNs) that contained different lesions. The modified ODNs were indirectly hybridized onto probes chemically fixed at defined sites on a circular format within each well of a 96-well microtiter plate (Oligo Sorbent Array, OLISA). The lesions were selected among oxidative damage (8-oxo-7,8 dihydroguanine, formylamine), deaminated bases (uracil, hypoxanthine) and alkylated base (N(6)-etheno-adenine). Cleavage specificity was checked using different enzymes: Fapy-DNA-N-glycosylase, 3-methyladenine DNA glycosylase II, uracil-N-glycosylase, endonuclease V and endonuclease VIII. The extent of excision could be monitored simultaneously for the selected base damage. For this purpose, we used automated fluorescence imaging analysis of the residual ODNs that contained lesions and remained on the support after release of the cleaved ODNs recognized by the repair enzymes. The results indicated that this assay could advantageously replace the analysis of glycosylase activities by PAGE techniques. Finally we show that this in vitro repair assay represents an interesting tool for the determination of cellular repair activities. PMID- 15351296 TI - A microtiter plate assay for the determination of the synthetic activity of protease. PMID- 15351297 TI - Anomalous mobility of polymerase chain reaction products after bisulfite treatment of DNA. PMID- 15351298 TI - Trehalose increases chemical-induced transformation efficiency of Escherichia coli. PMID- 15351299 TI - An effective substitute for triisopropylnaphthalenesulfonic acid in the preparation of plant RNA. PMID- 15351300 TI - The role of aquaporin-1 (AQP1) expression in a murine model of lipopolysaccharide induced acute lung injury. AB - A murine model of lipopolysaccharide (LPS)-induced acute lung injury (ALI) was used to evaluate whether aquaporin-1 (AQP1) is involved in lung inflammation and lung edema formation. Swiss strain mice (n = 122) had LPS (5 mg/kg) instilled intratracheally (IT), and were then treated with either 0.9 % saline or dexamethasone (5 mg/kg/day). Mice were euthanized at 2 days and 7 days after treatment. Inflammatory cytokines (TNF-alpha, IL-6), protein concentration in bronchoalveolar lavage (BAL) fluid, lung wet-to-dry weight ratio, histology, immunohistochemistry, and AQP1 Western blot were performed. Lung wet-to-dry weight ratio and lung vascular permeability were also measured in the AQP1 knockout mice (n = 9) that received IT LPS (5 mg/kg) at 2 days. Intratracheal instillation of LPS produced a severe lung injury at 2 days, characterized by elevation of TNF-alpha, IL-6 in the BAL fluid, and by histological changes consistent with increased lung vascular permeability and neutrophil infiltration. AQP1-immunoreactivity in the pulmonary capillary endothelium was reduced at 2 days and 7 days. Administration of dexamethasone improved LPS-induced ALI and retained expression of AQP1. However, depletion of AQP1 did not affect lung edema formation, lung vascular permeability, or lung histology. The results suggest that although AQP1 expression is decreased after lung injury, depletion of AQP1 does not alter lung inflammation and lung edema induced by LPS. PMID- 15351301 TI - Cardio-respiratory measures following isocapnic voluntary hyperventilation. AB - In some individuals, breathing is greater than at rest following voluntary hyperventilation. Most previous investigations have employed short hyperventilation periods; here we examine the time course of cardio-respiratory measures before, during, and after a 5-min voluntary hyperventilation, maintaining isocapnia throughout. We examined the possible co-involvement of the cardiovascular system; hypothesising that post-hyperventilation hyperpnoea results from an increase in autonomic arousal. In four subjects (two males, two females) of 18 (nine males, nine females) we observed a post-hyperventilation hyperpnoea, characterised by a slow decline of ventilation toward resting levels with a time constant of 109.0 +/- 16.1s. By contrast, heart rate, and systolic and diastolic blood pressure were unchanged from rest during and after voluntary hyperventilation for all subjects. We concluded that males and females were equally likely to exhibit post-hyperventilation hyperpnoea, and suggest that they may be characterised by an increased resting heart rate and the choice of breathing frequency to increase ventilation during the voluntary hyperventilation. We further concluded that post-hyperventilation hyperpnoea is rare, but when present is a strong and lasting phenomenon, and that it is not the result of an increased autonomic arousal. PMID- 15351302 TI - P2 receptors modulate respiratory rhythm but do not contribute to central CO2 sensitivity in vitro. AB - Multiple brainstem sites are proposed to contribute to central respiratory chemosensitivity, however, the underlying molecular mechanisms remain unknown. P2X2 subunit-containing ATP receptors, which mediate pH-sensitive currents, appear to contribute to central chemosensitivity in vivo [J. Physiol. 523 (2000) 441]. However, recent data from P2X2 knockout mice [J. Neurosci. 23 (2003) 11315] indicate that they are not essential. To further explore the role of P2 receptors in central chemosensitivity, we examined the effects of P2 receptor agonists/antagonists on respiratory-related activity and CO2-sensitivity of rhythmically-active in vitro preparations from neonatal rat. Our main findings: (i) that putative chemosensitive regions of the ventrolateral medulla are immunoreactive for the P2X2 subunit; (ii) that ATP potentiates respiratory frequency in a dose-dependent, and PPADS-sensitive (P2 receptor antagonist), manner; and (iii) that the increase in burst frequency produced by increasing CO2 is unaffected by PPADS, indicate that ATP is a potent modulator of respiratory activity, but that P2 receptors do not contribute to central chemosensitivity in vitro. PMID- 15351303 TI - Pontine respiratory group neuron discharge is altered during fictive cough in the decerebrate cat. AB - A network of neurons in the rostral dorsal lateral pons and pons/mescencephalic junction constitute the pontine respiratory group (PRG) and is essential for reflex cough. As a next step in understanding the role of the PRG in the expression of the cough reflex, we examined neuron firing rates during fictive cough in cats. Decerebrated, thoracotomized, paralyzed, cycle-triggered ventilated adult cats were used. Extracellular activity of many single neurons and phrenic and lumbar neurograms were monitored during fictive cough produced by mechanical stimulation of the intrathoracic trachea. Neurons were tested during control periods for respiratory modulation of firing rate by cycle-triggered histograms and statistical tests. Most respiratory modulated cells were continuously active with various superimposed respiratory patterns; major categories included inspiratory decrementing (I-Dec), expiratory decrementing (E Dec) and expiratory augmenting (E-Aug). There were alterations in the discharge patterns of respiratory, as well as, non-respiratory modulated neurons during cough. The results suggest an involvement of the PRG in the configuration of the cough motor pattern. PMID- 15351304 TI - Friction and lubrication of pleural tissues. AB - The frictional behaviour of rabbit's visceral pleura sliding against parietal pleura was assessed in vitro while oscillating at physiological velocities and amplitudes under physiological normal forces. For sliding velocities up to 3 cm s(-1) and normal compressive loads up to 12 cm H2O, the average value of the coefficient of kinetic friction (mu) was constant at 0.019 +/- 0.002 (S.E.) with pleural liquid as lubricant. With Ringer-bicarbonate solution, mu was still constant, but significantly increased (Deltamu = 0.008 +/- 0.001; P < 0.001). Under these conditions, no damage of the sliding pleural surfaces was found on light and electron microscopy. Additional measurements, performed also on peritoneum, showed that changes in nominal contact area or strain of the mesothelia, temperature in the range 19-39 degrees C, and prolonged sliding did not affect mu. Gentle application of filter paper increased mu approximately 10 fold and irreversibly, suggesting alteration of the mesothelia. With packed the red blood cells (RBC) between the sliding mesothelia, mu increased appreciably but reversibly on removal of RBC suspension, whilst no ruptures of RBC occurred. In conclusion, the results indicate a low value of sliding friction in pleural tissues, partly related to the characteristics of the pleural liquid, and show that friction is independent of velocity, normal load, and nominal contact area, consistent with boundary lubrication. PMID- 15351305 TI - Effect of facial cooling and cold air inhalation on sympathetic nerve activity in men. AB - In nine healthy subjects, cold stimuli were administered to the forehead and hand, to the oral and nasal cavities via ice cubes and to the bronchial system via inhalation of cold air (-25 degrees C). Blood pressure, heart rate and muscle sympathetic nerve activity (MSNA) from the peroneal nerve were recorded. MSNA expressed as total activity increased during cold air inhalation, cooling of the forehead (P < 0.001, ANOVA), hand and mouth (P < or = 0.05), paralleled by a rise in blood pressure during cold air inhalation and cooling of the forehead and hand (P < 0.01). Cooling of the forehead provoked a faster increase of MSNA expressed as total activity (P < 0.05) and higher levels of diastolic blood pressure (P = 0.05) compared with cooling of the hand. Bradycardia was observed only during cooling of the nasal cavity (P < 0.001) and the forehead (P < 0.05). It is concluded that cooling of the skin and mucous membranes of the tracheobronchial tract elicits sympathetically mediated hemodynamic adaptations, probably via stimulation of cold-sensitive afferents. PMID- 15351306 TI - Effects of mechanical load on flow, volume and pressure delivered by high frequency percussive ventilation. AB - High-frequency percussive ventilation (HFPV) has proved its unique efficacy in the treatment of acute respiratory distress, when conventional mechanical ventilation (CMV) has demonstrated a limited response. We analysed flow (V(dot)), volume (V) and airway pressure (Paw) during ventilation of a single-compartment mechanical lung simulator, in which resistance (R) and elastance (E) values were modified, while maintaining the selected ventilatory settings of the HFPV device. These signals reveal the physical effect of the imposed loads on the output of the ventilatory device, secondary to constant (millisecond by millisecond) alterations in pulmonary dynamics. V(dot), V and Paw values depended fundamentally on the value of R, but their shapes were modified by R and E. Although peak Paw increased 70.3% in relation to control value, mean Paw augmented solely 36.5% under the same circumstances (maximum of 9.4 cm H2O). Finally, a mechanism for washing gas out of the lung was suggested. PMID- 15351307 TI - Perspectives on the changing face of lupus mortality. PMID- 15351308 TI - Mortality studies in SLE: how far can we improve survival of patients with SLE. PMID- 15351309 TI - Lupus mortality in Japan. AB - Systemic lupus erythematosus (SLE)-related mortality has significantly declined in Japan during the past four decades. Decrease in number of deaths due to renal failure has considerably improved the prognosis of SLE. The adequate use of steroid therapy, the new advances in the second-line therapies including immunosuppressants, plasmapheresis and hemodialysis, and the appropriate control of infectious complications are the major reasons of the reduction of SLE-related mortality. The establishment of a nation-wide registration system of SLE in Japan will allow us to improve our strategy to control the disease. PMID- 15351310 TI - Surviving the butterfly and the wolf: mortality trends in systemic lupus erythematosus. AB - OBJECTIVE: To address changes in survival of patients with systemic lupus erythematosus (SLE) and to compare mortality statistics with associated disease specific as well as comorbid conditions. METHODS: Review of the international literature on survival of patients with SLE. RESULTS: During the first half of the 20th century, SLE was considered a rapidly progressive and almost invariably fatal disease. Since the 1950s, the estimated 5-year survival of SLE patients in developed countries rose from <50% to >95% and similar increases were seen in 10 year survival. Mortality rates of SLE patients, however, remain approximately 3 times that of an age- and sex-matched population in most studies, indicating that death still occurs prematurely in a substantial portion of patients, albeit later in the disease course. This improved prognosis does not appear to have been shared equally by all racial/ethnic groups. This appears to be attributable more to socioeconomic and sociocultural factors than to true differences in disease manifestations. Along with the increased survival of SLE patients, there has been a change in the causes of death. Most notably, there has been a dramatic increase in the proportionate mortality from vascular disease, particularly accelerated atherosclerosis. Both disease and therapeutic modalities, in particular corticosteroids, appear to contribute to the high prevalence of coronary artery disease (CAD). CONCLUSIONS: Much progress has been made in improving the survival of SLE, but there is need for further improvement. Aggressive treatment of risk factors for CAD is advisable, but it remains to be assessed to what extent such interventions can further reduce mortality. PMID- 15351311 TI - Possible roles and determinants of microchimerism in autoimmune and other disorders. AB - Microchimerism is the presence of a low level of non-host stem cells or their progeny in an individual. The most common source of microchimerism is pregnancy. During pregnancy, bi-directional trafficking of hematopoietic cells occurs through the placenta and these microchimeric cells persist for decades after childbirth. A possible role of microchimerism in the pathogenesis of some (systemic sclerosis, systemic lupus erythematosus, primary biliary cirrhosis, autoimmune thyroid diseases and juvenile myositis) but not all autoimmune diseases has been suggested by recent studies. Contradictory reports exist regarding HLA allelic associations with persistent T lymphocyte microchimerism. Although much of the focus of past studies has been on microchimerism in the effector arm of the immune system, increasing evidence suggests that microchimeric cells may differentiate into many lineages in different tissues raising additional possible roles for these cells. The possibility of microchimerism in many organs should induce an exploration of how persistent mixtures of cells of different genetic backgrounds throughout the body may influence diverse physiologic processes during life. In the present review, we discuss possible influencing factors and roles of all forms of microchimerism in autoimmune and non-autoimmune diseases. A better understanding of the immune mechanisms, along with the identification of environmental and genetic risk factors, is crucial for further deciphering the many possible implications of maternal-fetal and fetal-maternal cell trafficking in health and disease. PMID- 15351312 TI - Class switch recombination in B lymphopoiesis: a potential pathway for B cell autoimmunity. AB - Pathogenic autoantibodies detected in autoimmune diseases are predominantly IgG isotypes, reflecting the generation and activation of an autoimmune memory B cell repertoire. It is not completely understood how such autoreactive cells are generated and escape central and/or peripheral tolerance mechanisms, and several models to explain this have been proposed. It is generally thought that B lymphocytes utilize IgM receptors for development and tolerance establishment, whereas IgG receptors are primarily used to promote memory formation and signal for memory-type responses. In here we review recent findings suggesting that spontaneously occurring class switch recombination in B lymphopoiesis confer B lymphocytes with a novel developmental pathway that is driven by non-IgM receptors. The physiological relevance of this developmental pathway in generating an autoimmune memory repertoire, as well as a Fas-dependent mechanism regulating it, is discussed. PMID- 15351313 TI - NK95806, a newly synthesized microtubule-disrupting agent that suppresses collagen-induced arthritis in mice. AB - We investigated the anti-arthritic effects of NK95806, a novel inhibitor of microtubule polymerization, on collagen-induced arthritis in mice. The suppressive effect of NK95806 on the induction and development of arthritis was shown as a significant reduction in clinical arthritis scores. Histological analysis of the hind paws confirmed the improvement in clinical severity and showed marked decreases in granulomatous formation and further bone destruction. Further, under the experimental conditions in which methotrexate had little, if any, effect, NK95806 significantly suppressed the development of arthritis. These results suggest that the disruption of microtubules might be a novel target for anti-rheumatic drugs and NK95806 may be a candidate for further development. PMID- 15351314 TI - Protective function of complement against alcohol-induced rat liver damage. AB - The complement system can promote tissue damage or play a homeostatic role in the clearance and disposal of damaged tissue. We assessed the role of the terminal complement pathway in alcohol-induced liver damage in complement C6 (C6-/-) genetically deficient rats. C6-/- and corresponding C6+/+ rats were continuously exposed to ethanol by feeding ethanol-supplemented liquid diet for six weeks. Liver samples were analyzed for histopathology and complement component deposition by immunofluorescence microscopy. Prostaglandin E receptors and cytokine mRNA levels were analyzed by RT-PCR and plasma cytokines by ELISA. Deposition of complement components C1, C3, C8 and C9 was observed in C6+/+ rats, but not in C6-/- animals. The histopathological changes, the liver weight increase and the elevation of the plasma pro-/anti-inflammatory TNF-alpha/IL-10 ratio were, on the other hand, more marked in C6-/- rats. Furthermore, ethanol enhanced the hepatic mRNA expression of the prostaglandin E receptors EP2R and EP4R exclusively in the C6-/- rats. Our results indicate that a deficient terminal complement pathway predisposes to tissue injury and promotes a pro inflammatory cytokine response. This suggests that an intact complement system has a protective function in the development of alcoholic liver damage. PMID- 15351315 TI - An in vitro investigation of the effects of the nerve agent pretreatment pyridostigmine bromide on human peripheral blood T-cell function. AB - The current pretreatment against nerve agent poisoning deployed by the UK and US armed forces is the acetylcholinesterase (EC 3.1.1.7) inhibitor pyridostigmine bromide (PB). At higher doses, PB is also used to treat the autoimmune disease myasthenia gravis. In both cases, the therapeutic effect is mediated by inhibition of acetylcholinesterase (AChE) at cholinergic synapses. However, the location of AChE is not restricted to these sites. AChE, acetylcholine (ACh) receptors and choline acetyltransferase have been reported to be expressed by T cells, suggesting that cholinergic signalling may exert some modulatory influence on T-cell function and consequently on the immune system. The aim of this study was to investigate the role of the T-cell cholinergic system in the immunological activation process and to examine whether inhibitors of AChE such as PB affect immune function. To investigate this, human peripheral blood mononuclear cells (PBMC) were stimulated using either mitogen, cross-linking of the T-cell receptor and co-receptors with antibodies (anti-CD3/CD28) or by antigen presentation in the presence of various AChE inhibitors and ACh receptor agonists or antagonist. Several indices were used to assess T-cell activation, including the secretion of IL-2, cell proliferation and expression of CD69. Treatment with PB had no significant effect on the immunological assays selected. Physostigmine (PHY), a carbamate compound similar to PB, consistently showed inhibition of T-cell activation, but only at concentrations in excess of those required to inhibit AChE. No evidence was found to support previously published findings showing muscarinic enhancement of cell proliferation or IL-2 secretion. PMID- 15351316 TI - Dehydroepiandrosterone induces apoptosis of thymocyte through Fas/Fas-L pathway. AB - Dehydroepiandrosterone (DHEA), the most plentiful steroid hormone, has been convincingly known to have many biological effects and diverse influence in various types of cells, tissues and organs. The effects of DHEA on the humoral and cellular immune response are widely tested, but it is unclear whether DHEA itself serves as an activator of immune function and the literature pertaining to the in vitro effects of DHEA remains contested. In the present paper, the effects of DHEA on the thymocytes in vitro were studied. The results showed that DHEA could enhance the expression of Fas and Fas-L and induce thymocyte apoptosis. This suggests that dehydroepiandrosterone may induce the apoptosis of thymocyte through Fas/Fas-L pathway. PMID- 15351317 TI - Activation of mitogen-activated protein kinases and AP-1 by polysaccharide isolated from the radix of Platycodon grandiflorum in RAW 264.7 cells. AB - The root of Platycodon grandiflorum has been widely used for the treatment of various diseases in oriental medicine. Our previous study showed that the PG, a polysaccharide isolated from P. grandiflorum, activates macrophages via Toll-like receptor 4 (TLR4). However, the associated biological mechanisms are not fully understood. To elucidate the molecular mechanism responsible for the macrophage activation, we investigated the effect of PG on the activity of mitogen-activated protein kinases (MAPKs) and activator protein-1 (AP-1) in RAW 264.7 cells, a murine macrophage cell line. Treatment of RAW 264.7 cells with PG produced a marked induction of AP-1 DNA binding activity. Moreover, all three MAPKs were activated by PG, and PG-induced activation of MAPKs was abrogated by the treatment of PD98059, curcumin, and SB203580, specific inhibitors of MEK-1/2, stress-activated protein kinases/jun N-terminal kinase (SAPK/JNK), and p38 MAP kianse, respectively. The induction of AP-1 DNA binding activity by PG was also inhibited by these MAPK inhibitors. Moreover, supershift analysis identified that JunB and Fra-1 are major components involved in the PG-mediated induction of AP-1 DNA binding. Additionally, curcumin and SB203580 suppressed PG-induced production of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), whereas PD98059 showed an inhibitory effect only on the TNF-alpha production. Taken together, these results suggest that macrophage activation by PG is mediated, at least in part, by MAPKs and AP-1. PMID- 15351318 TI - Methanol extract of Dioscoreae Rhizoma inhibits pro-inflammatory cytokines and mediators in the synoviocytes of rheumatoid arthritis. AB - Dioscoreae Rhizoma (MDR), the root of Dioscorea tokoro MAKINO, has been used for the treatment of arthritis, muscular pain and urinary diseases in oriental medicine. The present work evaluates a methanol extract of Dioscoreae Rhizoma (MDR). MDR did not show any cytotoxic effect on mouse lung fibroblast cells (mLFCs) or human fibroblast-like synovial cells (hFLSCs). However, it significantly reduced the proliferation of hFLSCs stimulated by interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha). MDR significantly inhibited the production of TNF-alpha and IL-1beta as well as down-regulating the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in IL-1beta- and TNF-alpha-stimulated hFLSCs. MDR also effectively reduced the level of reactive oxygen species (ROS) in these cells. Taken together, these findings provide evidence that MDR may be a candidate for the treatment of rheumatoid arthritis (RA). PMID- 15351319 TI - Characterization of anti-inflammatory compounds using transcriptomics, proteomics, and metabolomics in combination with multivariate data analysis. AB - The discovery of new anti-inflammatory drugs is often based on an interaction with a specific target, although other pathways often play a primary or secondary role. Anti-inflammatory drugs can be categorized into classes, based on their mechanism of action. In this article we investigate the possibility to characterize novel anti-inflammatory compounds by three holistic methods. For this purpose, we make use of macrophage-like U937 cells which are stimulated with LPS in the absence or presence of an anti-inflammatory compound. Using micro arrays, 2-D gel electrophoresis and a LC-MS method for lipids the effects on the transcriptome, proteome and metabolome of the exposed cells is investigated. The expression patterns are subsequently analyzed using in-house developed pattern recognition tools. Using the methods described above, we have examined the effects of six anti-inflammatory compounds. Our results demonstrate that different classes of anti-inflammatory compounds show distinct and characteristic mRNA, protein, and lipid expression patterns, which can be used to categorise known molecules and to discover and classify new leads. The potential of our approach is illustrated by the analysis of several beta (2)-adrenergic agonists (beta2-agonists). In addition to their primary pharmacological target, beta2 agonists posses certain anti-inflammatory properties. We were able to show that zilpaterol, a poorly characterized beta2-agonist, gives rise to an almost identical expression pattern as the beta2-agonists clenbuterol and salbutamol. Furthermore we have identified specific mRNA, protein and lipid markers for the anti-inflammatory compounds investigated in this study. PMID- 15351320 TI - The impact of a new immunomodulator oxo-quinoline-3-carboxamide on the progression of experimental lupus. AB - Autoimmune, lupus-prone MRL lpr/lpr mice were treated orally with oxo-quinoline-3 carboxamide (ABR-25757), a newly developed immunomodulator. Treatment was initiated in one set of experiment at the age of 10 weeks, before the onset of clinically apparent disease, and in another set at 15 weeks, after the development of established lupus disease. Beneficial therapeutic effects were obtained even when ABR-25757 was administered at the lowest dose tested (7.5 microg/mouse/week) to 15 weeks old mice with established lupus disease. The effects of ABR-25757 on longevity, as well as on development of glomerulonephritis were pronounced and comparable with those of LS-2616, a potent immunomodulator. Administration of ABR-25757 did not significantly alter T cell responses in vivo nor in vitro. In addition, it only marginally suppressed B cell responses measured as frequencies of immunoglobulin secreting cells. By the same token this compound did not affect overall leukocyte content in primary (bone marrow) or secondary (spleen) lymphoid tissues. In contrast, treatment with ABR 25757 up regulated expression of pro-inflammatory transcription factors NF-kappaB and AP-1. These results suggest (a) a potential therapeutic role of ABR-25757 in the treatment of experimental lupus and (b) that the effect of the treatment is mediated by immunodeviation rather than by immunosuppression. PMID- 15351321 TI - (S)-(+)-methadone is more immunosuppressive than the potent analgesic (R)-(--) methadone. AB - Methadone is a widely used synthetic opioid which is administered as a racemic mixture of (R)-(--)- and (S)-(+)-enantiomers, with only (R)-(--)-methadone possessing mu opioid receptor agonist activity. Methadone inhibits numerous immune functions in vitro at concentrations above 10 microM in a nonstereoselective and naloxone-insensitive fashion, suggesting the presence of nonclassical opioid receptors on immune cells. No in vivo data on the effects of methadone's enantiomers on immune function are available. Therefore, the stereoselectivity of methadone's analgesia (hot plate latency) in vivo and immune suppression ex vivo (splenocyte proliferation) was investigated in groups of Balb/c mice. Significant analgesia was observed in animals that received racemic methadone (P=0.0012, 52% MPE) and (R)-(--)-methadone (P=0.0002, 70% MPE) when compared to saline-treated controls, while (S)-(+)-methadone was devoid of any such effect (-4% MPE). In vivo (R)-(--)- and racemic methadone caused significant inhibition (P<0.001, greater than -70%) of basal proliferation compared to saline control. In stark contrast to analgesia, in vivo (S)-(+)-methadone caused significantly greater inhibition of basal proliferation (P<0.001, -130%) than (R) (--)- and racemic methadone. The immune suppression caused by methadone is not purely a classical opioid response but involves nonclassical opioid receptors located at the central level, which have yet to be characterised. Moreover, the dose at which immune suppression occurred could be achieved clinically. PMID- 15351322 TI - Participation of chemical mediators in the development of experimental allergic conjunctivitis in rats. AB - In the present study, we investigated the participation of chemical mediators in the development of experimental allergic conjunctivitis in rats. Cetirizine (a histamine H1 receptor antagonist), ramatroban (a thromboxane A2 (TXA2) receptor antagonist) and zafirlukast (a cysteinyl leukotrienes (cys-LTs) receptor antagonist) were orally administered from day 14 to day 42 during repeated topical antigen challenge. An increase in reactivity to antigen- and histamine induced eye scratching behavior was observed by topical sensitization in sensitized rats. Although increased reactivity to antigen was not influenced by cetirizine, ramatoroban and zafirlukast, increased reactivity to histamine was significantly inhibited by ramatroban. The development of conjunctival edema was also observed for topical sensitization. Cetirizine caused no inhibition of the development of conjunctival edema, but ramatroban and zafirlukast inhibited the development of conjunctival edema. In addition, the number of eosinophils in the conjunctiva was increased by topical sensitization. Cetirizine had no significant effect on the increase in the number of eosinophils. However, ramatroban and zafirlukast were effective in inhibiting an increase in the number of eosinophils induced by topical sensitization. These results indicate that TXA2 is involved in increased histamine reactivity, and TXA2 and cys-LTs are associated with not only the conjunctival edema but also eosinophil infiltration during the development of experimental allergic conjunctivitis in rats. PMID- 15351323 TI - Angiogenesis as an immunopharmacologic target in inflammation and cancer. AB - Many pathological processes including wound healing, chronic inflammation and cancer require angiogenesis, i.e., the formation of new vasculature in the lesions. Accumulating evidence indicates that angiogenesis is crucial for both chronic inflammation and the growth of malignant tumors with the participation of diverse cytokines, chemokines and growth factors. It is nevertheless believed that differences exist in angiogenesis between cancer and chronic inflammatory diseases. The aim of this review is to outline the characteristics of angiogenesis in chronic inflammation and cancer. A better understanding of the angiogenic processes may facilitate the design of more effective therapies for chronic inflammation and cancer. PMID- 15351324 TI - Differential effect of Coriolus versicolor (Yunzhi) extract on cytokine production by murine lymphocytes in vitro. AB - Being one of the commonly used Chinese medicinal herbs, Coriolus versicolor (CV), also named as Yunzhi, was known to possess both anti-tumor and immunopotentiating activities. The present study aimed to investigate the in vitro immunomodulatory effect of a standardized ethanol-water extract prepared from CV on the proliferation of murine splenic lymphocytes using the MTT assay, and the production of six T helper (Th)-related cytokines using the enzyme-linked immunosorbent assay (ELISA) technique. The results showed that the CV extract significantly augmented the proliferation of murine splenic lymphocytes in a time and dose-dependent manner, maximally by 2.4-fold. Moreover, the production of two Th1-related cytokines, including interleukin (IL)-2 and IL-12, in culture supernatants from the CV extract-activated lymphocytes was prominently upregulated at 48 and 72 h. Positive correlations were found between the levels of these two cytokines and the MTT-based proliferative response. In contrast, the production of two other Th1-related cytokines, including interferon (IFN)-gamma and IL-18, was significantly augmented only at 24 h, but not at 48 and 72 h. On the other hand, the levels of two Th2-related cytokines such as IL-4 and IL-6 were undetectable in the culture supernatants of lymphocytes treated with the CV extract. The CV extract was suggested to be a lymphocyte mitogen by differentially enhancing the production of Th1-related cytokines. PMID- 15351325 TI - Parenteral immunization to beta-lactoglobulin modifies the intestinal structure and mucosal electrical parameters in rabbit. AB - Systemic and local immune responses and the intestinal structure were examined in parenterally beta-Lg-sensitized rabbits. Immunization led to high IgG titers against beta-Lg. In a Ussing chamber, a sensitized ileum had a higher short circuit current (Isc) and potential difference (PD) than a control following in vitro beta-Lg challenge. Histological study indicated that presence of the sensitizing antigen affected and considerably modified the structure of the intestinal mucosa in sensitized rabbits when compared to controls. These alterations were revealed by active atrophy and marked infiltration of the lymphocytes. These findings indicate that antigen exposure results in morphological changes and abnormalities affecting the transport of water and electrolytes. This study provides a clearer understanding of the physiopathological mechanisms of allergy to cow's milk protein. PMID- 15351326 TI - The spirit of giving. PMID- 15351327 TI - The theoretical basis for nurse-midwifery practice in the United States: a critical analysis of three theories. AB - The relatively recent effort of midwifery scholars has resulted in the development of three middle-range midwifery theories in the United States. This article examines the three theoretical models that have been developed to describe the essential components of midwifery practice. The three theorists demonstrate remarkable consistency in the identification of concepts important to the discipline, which includes the following essential characteristics of the midwifery paradigm of care: 1) acknowledgment of connections between the mind and body and the person to the person's life and world; 2) assuming the perspective of the woman to investigate meaning and her experience of symptoms or conditions, so that a plan of care is developed by midwife and woman together; and 3) protection and nurturance of the "normal" in processes related to women's health, implying a judicious use of technology and intervention. PMID- 15351328 TI - First birth stories of student midwives: keys to professional affective socialization. AB - The purpose of this article was to examine the professional affective socialization of midwifery students. Students from the Community-Based Nurse Midwifery Education Program (CNEP) tell about their first births in their Lost My Hat stories posted on the electronic bulletin board of the Frontier School of Midwifery and Family Nursing. Qualitative analysis of 38 of these stories showed internalization of the values and beliefs of the midwifery model of care by students. Themes of the Lost My Hat stories were congruent with some of the aspects of the Model of Exemplary Midwifery Practice developed by Kennedy. Major themes included the circle of safety provided by the preceptor and a woman centered approach to care. These findings can help midwifery educators promote socialization of students in the profession. PMID- 15351329 TI - Redesigning prenatal care through CenteringPregnancy. AB - CenteringPregnancy is a model of group prenatal care that provides more than 20 hours of contact time between the childbearing care provider and a cohort of pregnant women with similar due dates. During this time, each woman has the opportunity to build community with other pregnant women, learn self-care skills, get assurance about the progression of her pregnancy, and gain knowledge about pregnancy, birth, and parenting. Ten essential elements have been defined, which contribute to the success of this model of prenatal care delivery. These elements correspond with the Institute of Medicine's 2001 challenge to improve the quality of health care in the United States. Foundational perspectives provide potential explanations for the model's growing influence and success. Implications for clinical practice and further research to link it with perinatal health outcomes are suggested. PMID- 15351330 TI - CenteringPregnancy and the current state of prenatal care. AB - Prenatal care is often credited with improving pregnancy outcomes. Yet rates of low birthweight (LBW) and prematurity have risen in recent decades, calling into question the efficacy of traditional prenatal routines. Proposals have included broadening the objectives of prenatal care beyond prevention of LBW and enriching care to provide education and support for pregnant women. CenteringPregnancy, an innovative model of prenatal care that integrates extensive health education and group support with the standard prenatal exam, incorporates many of these elements. Impediments to wider implementation of CenteringPregnancy are explored, as well as proposals for addressing these challenges. PMID- 15351331 TI - Pregnancy outcomes of adolescents enrolled in a CenteringPregnancy program. AB - Adolescent pregnancy remains a significant social, economic, and health issue in the United States. The unique developmental needs of the pregnant adolescent require attention when designing prenatal care services. The CenteringPregnancy model of group prenatal care provides education and support for young women in an active and developmentally appropriate environment. Thirteen groups of adolescents (N = 124) have completed the Centering program at the Teen Pregnancy Center at Barnes Jewish Hospital in St. Louis, Missouri. Evaluation data suggest that the model has encouraged excellent health care compliance, satisfaction with prenatal care, and low rates of preterm birth and low birth weight infants. PMID- 15351332 TI - Elective cesarean birth: issues and ethics of an informed decision. AB - Is elective cesarean birth an issue of a woman's autonomy to choose her mode of birth or fear of labor by both the patient and the providers? Are women consenting to this procedure truly informed about the short-term and long-term risks associated with cesarean birth versus vaginal birth? This article examines the issues associated with elective cesarean birth and the factors that have led to recent consideration of this option. It is essential that we evaluate how risks, benefits, and liability issues impact the recommendations for the mode of birth. The ethics of making an informed choice as well as ethical professional responsibilities are explored. PMID- 15351333 TI - Postpartum sexual functioning and method of delivery: summary of the evidence. AB - Short-term postpartum sexual problems are highly prevalent, ranging from 22% to 86%; however, there are few studies that address how mode of delivery affects sexual functioning after childbirth. The objective of this study was to perform a systematic review of the literature on selected postpartum sexual function outcomes as affected by cesarean, assisted vaginal, and spontaneous vaginal delivery. We searched PubMed, CINAHL, and Cochrane databases from January 1990 to September 2003 and focused on mode of delivery and the most commonly reported sexual health outcomes, which included perineal pain, dyspareunia, resumption of intercourse, and self-reported perception of sexual health/sexual problems. The studies all showed increased risks of delay in resumption of intercourse, dyspareunia, sexual problems, or perineal pain associated with assisted vaginal delivery. Some studies showed no differences in sexual functioning between women with cesarean delivery and those with spontaneous vaginal delivery, whereas others reported less dyspareunia for women with cesarean delivery. A systematic review of the literature suggests an association between assisted vaginal delivery and some degree of sexual dysfunction. Reported associations between cesarean delivery and sexual dysfunction were inconsistent. Continued research is necessary to identify modifiable risk factors for sexual problems related to method of delivery. PMID- 15351334 TI - Caring for women from culturally diverse backgrounds: midwives' experiences. AB - The aim of this study was to show how midwives cared for women from culturally diverse backgrounds. In-depth interviews were used to collect data from 12 experienced midwives who volunteered to participate in the study from a midwifery unit with a culturally diverse population. Study findings revealed that midwives preserved and accommodated the cultural preferences of women from a Chinese background by incorporating the forces of yin-yang into care, heeding the maternal hierarchy and women's stoicism; and for women from an Islamic background by heeding modesty and gender preferences (Hejab), the place of prayer in daily life (Salat), and the imperative of visiting by others (Hadith). Hence, midwives negotiated care that was culturally comfortable for women and their families. Furthermore, triangulated studies addressing the partnership between the midwife and the diverse client are needed, as well as the development of aspects of the health service that are more culturally sensitive. PMID- 15351335 TI - Do birth certificate data accurately reflect the number of CNM-attended births? An exploratory study. AB - The number of midwife-attended births is increasing as reported on birth certificates in the United States. However, there is some evidence that births attended by certified nurse-midwives (CNMs) may not be accurately recorded. In this exploratory study, data on birth attendants for those clients giving birth during the study period were compared by using four sources: the client's hospital chart, the CNM birth log, hospital birth certificate records, and state vital statistics records. Researchers sought to determine the accuracy of birth attendant data as reflected in these four sources and whether other providers were listed as the birth attendant for actual CNM-attended births. During the study period, the CNM birth log showed that CNMs attended 97 vaginal births, whereas the client hospital charts for these same births noted 92 births as attended by CNMs (the other five were operative vaginal births). Hospital birth certificate and state vital statistics data during the study time period credited 88 and 82 of the client's births, respectively, to the CNMs. Exploration of the inaccurately reported birth attendant data, implications for practice, and recommendations for accurately recording birth certificate data are discussed. PMID- 15351336 TI - Problem-based learning in reproductive physiology. AB - One of the most misunderstood concepts in progressive educational curricula is that of problem-based learning (PBL). Often confused with problem solving, the two are quite different. Courses conducted in a PBL format use problems as a launch pad for resulting study. This article reviews a course designed to enable students to learn and understand the principles of human reproductive physiology using PBL. The learning process is situation based, student directed, and takes place in small groups. The rationale for some of these innovations is discussed and the way in which they were implemented is described. PMID- 15351337 TI - Is domperidone safe for breastfeeding mothers? PMID- 15351340 TI - Defining the problem: psychiatric and behavioral comorbidity in children and adolescents with epilepsy. AB - A variety of comorbid psychiatric conditions are frequently identified in children and adolescents with epilepsy, including depression, anxiety, psychosis, and attention-deficit hyperactivity disorder. Data regarding the epidemiology and precise prevalence of comorbid disorders in childhood epilepsy are incomplete and just now beginning to be compiled. Psychiatric and behavioral comorbidities are believed to affect approximately 40-50% of children and adolescents with epilepsy. Optimal diagnosis, clinical evaluation, and choice of treatment are predicated on the proper identification of coexisting psychiatric and behavioral disorders. Comorbid conditions in children and adolescents with epilepsy should be evaluated and treated as soon as they are recognized. PMID- 15351341 TI - Differential diagnosis and treatment of psychiatric disorders in children and adolescents with epilepsy. AB - Behavioral problems, such as the inattentive form of attention-deficit hyperactivity disorder (ADHD), anxiety, and depression, are common in children and adolescents with epilepsy and especially associated with central nervous system damage, family dysfunction, and severe seizures. This article discusses the risk factors to be considered when focusing on the prevalence of behavioral problems, the family factors that influence their incidence, as well as the differential diagnosis of behavioral disorders commonly associated with epilepsy. It also considers the assessment of these behavioral disorders and their treatment with psychotherapy, education, and a variety of psychopharmacological agents. PMID- 15351342 TI - Effects of epilepsy surgery on psychiatric and behavioral comorbidities in children and adolescents. AB - When considering surgery in children, it is important to think about the potential effect on psychiatric and behavioral outcome. A growing body of evidence suggests that the type of epilepsy (benign or catastrophic) and the age of the patient affect developmental outcome. In younger children with catastrophic epilepsy, the primary goal of therapy is not only to control seizures, but also to offer the child the opportunity for the best possible development. In older children with partial seizures, the therapeutic goal is to control seizures, which may allow a more normal life and improve behavior. Epilepsy surgery generally appears to have a positive effect on behavior and development in many younger children with catastrophic epilepsy. Psychiatric and behavioral outcomes in older children with complex partial epilepsy are less clear; many patients improve, but there does appear to be a small risk of developing new psychiatric or behavioral disorders postoperatively. PMID- 15351343 TI - Effects of antiepileptic medications on psychiatric and behavioral comorbidities in children and adolescents with epilepsy. AB - The three goals of this article are (1) to delineate the limitations in determining the actual incidence of antiepileptic drug (AED) psychiatric and behavioral side effects; (2) to summarize existing data on the direct effects of AEDs on psychiatric and behavioral comorbidities and examine the relationship between these direct effects and specific AED mechanisms of action; and (3) to recognize the indirect effects of AEDs on psychiatric and behavioral medications that can result in aggravation of these comorbidities through drug-drug interactions. All of these data are then combined and formatted into a practical algorithm useful in many clinical situations. PMID- 15351344 TI - Behavioral issues involving children and adolescents with epilepsy and the impact of their families: recent research data. AB - OBJECTIVE: Using data from a larger study on new-onset seizures, we reported preliminary findings concerning relationships between family factors and child behavioral problems at baseline and 24 months. We also explored which baseline and changes in family factors were associated with changes in child behavioral problems over the 24-month period. METHODS: Subjects were 224 children and their primary caregivers. Data were collected using structured telephone interviews and analyzed using multiple regression. RESULTS: Deficient family mastery and parent confidence in managing their child's discipline were associated with behavior problems at baseline and at 24 months; they also predicted child behavior problems over time. Decreasing parent confidence in disciplining their child was associated with increasing child behavior problems. Decreases in parent emotional support of the child were associated with increases in child internalizing problems. CONCLUSION: Child behavior problems, family environment, and parenting behaviors should be assessed when children present to the clinical setting with new-onset seizures. PMID- 15351346 TI - New nucleoside reverse transcriptase inhibitors for the treatment of HIV infections. AB - Several new nucleoside analogs are currently in development for the treatment of HIV-1 infections. Alovudine, amdoxovir, elvucitabine, Racivir, Reverset and SPD 754 are nucleoside reverse transcriptase inhibitors that were designed and selected in anticipation of having improved resistance, safety, compatibility and efficacy profiles. Clinical trials are demonstrating that some of these goals are being met, and that nucleoside analogs as a class of compounds remain fertile ground for finding valuable additions to current anti-retrovirus treatment regimens. PMID- 15351347 TI - New non-nucleoside reverse transcriptase inhibitors (NNRTIs) in development for the treatment of HIV infections. AB - Despite the availability of 20 approved anti-retroviral drugs for the treatment of HIV infection, there is still a need for new anti-retrovirals to improve convenience, reduce toxicity and, of particular importance, to provide activity against the growing number of drug-resistant HIV strains. A new generation of potent HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs) is emerging that inhibit HIV-1 strains resistant to the NNRTIs that are prescribed today, and which provide a higher genetic barrier for resistance development than do their predecessors. Of several NNRTIs that are in preclinical and clinical development, two agents, capravirine and TMC125, have shown promise in early clinical trials. The persistent and systematic study of the capacity of HIV to evolve under drug pressure, combined with basic studies in the mode of action of NNRTIs, can provide new weapons in the fight against AIDS. PMID- 15351348 TI - The current status of, and challenges in, the development of CCR5 inhibitors as therapeutics for HIV-1 infection. AB - The discovery of CCR5 as a HIV-1 co-receptor unfolded the cryptic and complicated process of HIV-1 cellular entry and has provided more than a few entry steps as possible modalities for effective viral intervention. The proof-of-principle has already been established for the use of entry inhibitors against HIV-1 and there is a cautious optimism that several CCR5 inhibitors might soon be added to our armamentarium for therapy of HIV-1 infection. PMID- 15351349 TI - Antivirals against DNA viruses (hepatitis B and the herpes viruses). AB - Antiviral drugs against DNA viruses are widely used for the management of diseases caused by infections with the Herpes viruses and have recently been introduced for Hepatitis B. There are also several emerging treatments (i.e. those that are in clinical development) and novel treatments that are still in the preclinical phase. Although the majority of emerging drugs are nucleoside analogues, there is a trend towards the development of non-nucleosidic drugs with unique mechanisms of action, in the hope that efficacy will be maximised and drug resistance and viral rebound minimised. PMID- 15351350 TI - Strategies for hepatitis C therapeutic intervention: now and next. AB - Interferon-alpha-based therapies are the mainstay of hepatitis C treatment. However, these broad immunomodulatory agents, already limited by their side effects, expense and route of administration, are not efficacious in all cases. Therefore, research efforts have focused on identifying additional drugs and therapeutic modalities to benefit those individuals refractive to current therapy, and which might be more affordable, such as orally active small-molecule alternatives and preventive or therapeutic vaccines. It is likely that successful therapeutic intervention of hepatitis C will require a combination of approaches targeting different aspects of the viral life-cycle. PMID- 15351351 TI - Glycopeptides in clinical development: pharmacological profile and clinical perspectives. AB - Vancomycin and teicoplanin are the two glycopeptides currently used in the clinics for the treatment of multiresistant infections by Gram-positive organisms. The development of resistance in enterococci and staphylococci has stimulated the search for new derivatives with improved activity, particularly against strains resistant to conventional derivatives. Three of these, obtained by hemi-synthesis starting from natural compounds, are now in clinical development (oritavancin and telavancin, as derivatives of vancomycin; and dalbavancin, as a derivative of teicoplanin). The presence of a lipophilic tail on these molecules results in them having a prolonged half-life. It also modifies their mode of action, conferring to them a concentration-dependent bactericidal activity. Their spectrum of activity includes methicillin-susceptible or methicillin-resistant staphylococci, penicillin-resistant pneumococci and enterococci (including vancomycin-resistant strains for oritavancin and telavancin). Ongoing clinical studies are evaluating the efficacy and safety of these molecules for the treatment of complicated skin and soft tissue infections and bactereamia, in a once-daily (oritavancin, telavancin) or once-weekly (dalbavancin) scheme of administration. Despite these remarkable properties, the use of these potent molecules should be restricted to severe infections by multiresistant organisms to limit the risk of selection of resistance. PMID- 15351352 TI - Structural understanding of efflux-mediated drug resistance: potential routes to efflux inhibition. AB - The active efflux of cytotoxic drugs mediated by multidrug transporters is the basis of multidrug resistance in prokaryotic and eukaryotic cells. Individual multidrug transporters can be extremely versatile, often exhibiting a staggering range of substrate specificity that can negate the effects of clinically relevant therapies. The effective treatment of bacterial, fungal and protozoan infections, along with certain cancer treatments, has been compromised by the presence of multidrug transporters. Traditionally, advances in the understanding of multidrug transporters have been made through biochemical analyses; more recently, however, fundamental advances have been made with the elucidation of several three dimensional structures of representative multidrug pumps. Biochemical and structural analysis of multidrug pumps could lead to the development of novel 'anti-efflux' therapies. PMID- 15351354 TI - High-throughput X-ray crystallography for drug discovery. AB - Knowledge of the three-dimensional structures of protein targets has the potential to greatly accelerate drug discovery, but technical challenges and time constraints have traditionally limited its use to lead optimization. Its application is now being extended beyond structure determination into new approaches for lead discovery. Structure-activity relationships by nuclear magnetic resonance have been widely used to detect ligand binding and to give some indication of the location of the binding site. X-ray crystallography has the advantage of defining ligand-binding sites with greater certainty. High throughput approaches make this method applicable to screening to identify molecular fragments that bind protein targets, and to defining precisely their binding sites. X-ray crystallography can then be used as a rapid technique to guide the elaboration of the fragments into larger molecular weight compounds that might be useful leads for drug discovery. PMID- 15351355 TI - Identification of CFTR activators and inhibitors: chance or design? AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP activated Cl(-) channel expressed in various epithelial cells, and is a pharmacological target for activators and inhibitors. Activators are useful for the pharmacotherapy of cystic fibrosis, specifically for those mutations that affect CFTR protein by reducing its ability to stay in the open state. Conversely, inhibitors are potentially useful to treat secretory diarrhoea caused by enterotoxins, as the CFTR is the main route for Cl(-) flux in the intestine. Recently, a variety of potent modulators of the CFTR Cl(-) channel activity have been identified by high-throughput screening of a large collections of small molecules. The identification of CFTR activators and inhibitors with novel chemical scaffolds might help with the rational design of compounds with improved pharmacological properties. PMID- 15351356 TI - Zebrafish as a pharmacological tool: the how, why and when. AB - Zebrafish combine the relevance of a vertebrate with the scalability of an invertebrate. They can live in 96-well plate format and readily absorb chemicals from the water. These features have stimulated the use of zebrafish by medical researchers to model human disease and then assess the action of compounds in a whole organism. Examples of the power of this system have been illustrated with the cloning of zebrafish human ether-a-go-go-related gene (HERG), which shows near 100% homology in key domains, and the associated ability to identify drugs that prolong the QT interval both rapidly and with tiny amounts (micrograms) of compound. PMID- 15351357 TI - Therapeutic targets from a Drosophila model of Alzheimer's disease. AB - The dominating role of the mouse in modeling Alzheimer's disease has been challenged this year by Drosophila melanogaster. Transgenic flies expressing toxic beta-amyloid peptides develop neurodegeneration. It is these peptides that accumulate in human disease and are thought to be the initiating factor in Alzheimer's disease. The flies exhibit a clear phenotype from a few days of age, including reduced locomotor function, impaired olfactory memory and shortened lifespan. Therapeutic agents that interfere with the generation of toxic aggregates of beta-amyloid peptides have been shown to rescue the flies. Several groups are now using the power and speed of genetic screens in the fly to accelerate the discovery of novel therapeutic agents. PMID- 15351358 TI - The application of functional magnetic resonance imaging to neuropharmacology. AB - The technique of functional magnetic resonance imaging (fMRI) has the capacity to acquire data with spatial and temporal resolution that far exceeds other currently available methods of non-invasive investigation of brain function. This coupled with its ability for serial studies makes it an attractive prospect for investigating the effects of pharmacological agents in the brain. Recent advances in fMRI have been made in the areas of reward and dependence, brain trauma and injury, psychotropic drugs and pain using small animals. Although the use of fMRI in pharmacological studies is becoming popular, there are various associated complications, such as the possible interference of drugs with the mechanisms that give rise to the pharmacological fMRI signal, and local or global cardiovascular changes that might produce functional responses unrelated to neural activity. Consideration of these concerns, coupled with careful attention to experimental detail and verification procedures, promises to make pharmacological fMRI use a valuable tool for understanding the actions of drugs in the brain. PMID- 15351359 TI - siRNA for gene silencing: a route to drug target discovery. AB - The identification of RNA interference in mammalian cells, mediated via both virally-derived short interference RNA (siRNA) and endogenously produced microRNA, has revolutionised our understanding of the translational control of gene expression. Indeed, since its initial discovery, siRNA has been rapidly deployed for the elucidation of gene function and the identification of potential drug targets, a process often known as target discovery. In this review, we briefly discuss the mechanism of RNA interference and then critically examine the use of siRNA in target discovery, with a particular emphasis upon issues such as efficacy, selectivity, delivery and application in high-throughput studies. PMID- 15351360 TI - Pharmacological chaperone action on G-protein-coupled receptors. AB - An increasing number of genetic diseases are found to result from mutations that lead to retention of the affected proteins in the endoplasmic reticulum, where they are recognized as misfolded by the quality control system. Several of these conformational diseases involve mutations in G-protein-coupled receptors. Recent studies demonstrated that pharmacologically selective compounds, termed pharmacological chaperones, can stabilize the misfolded receptors, facilitating their export from the endoplasmic reticulum to the plasma membrane, where they can be active. Such functional rescue suggests that pharmacological chaperones could represent novel therapeutic agents for the treatment of conformational diseases. Although only a few examples are currently available, the observation that pharmacological chaperones can also favour the folding of wild-type G protein-coupled receptors indicates that these compounds could have wide applications. PMID- 15351361 TI - EEG source imaging. AB - OBJECTIVE: Electroencephalography (EEG) is an important tool for studying the temporal dynamics of the human brain's large-scale neuronal circuits. However, most EEG applications fail to capitalize on all of the data's available information, particularly that concerning the location of active sources in the brain. Localizing the sources of a given scalp measurement is only achieved by solving the so-called inverse problem. By introducing reasonable a priori constraints, the inverse problem can be solved and the most probable sources in the brain at every moment in time can be accurately localized. METHODS AND RESULTS: Here, we review the different EEG source localization procedures applied during the last two decades. Additionally, we detail the importance of those procedures preceding and following source estimation that are intimately linked to a successful, reliable result. We discuss (1) the number and positioning of electrodes, (2) the varieties of inverse solution models and algorithms, (3) the integration of EEG source estimations with MRI data, (4) the integration of time and frequency in source imaging, and (5) the statistical analysis of inverse solution results. CONCLUSIONS AND SIGNIFICANCE: We show that modern EEG source imaging simultaneously details the temporal and spatial dimensions of brain activity, making it an important and affordable tool to study the properties of cerebral, neural networks in cognitive and clinical neurosciences. PMID- 15351362 TI - Differential modulation of the short- and long-latency somatosensory evoked potentials in a forewarned reaction time task. AB - OBJECTIVE: We investigated modulation of the short- and long-latency somatosensory evoked potentials (SEPs) in a forewarned reaction time task. METHODS: A pair of warning (auditory) and imperative stimuli (somatosensory) was presented with a 2 s interstimulus interval. In movement condition, subjects responded by grip movement with the ipsilateral hand to the somatosensory stimulation when the imperative stimulus was presented. In counting condition, they silently counted the number of imperative stimuli. The SEPs in response to the imperative stimuli were recorded. RESULTS: Frontal N30 and central N60 amplitudes were significantly smaller in the movement than in the counting or rest conditions. None of the short-latency components differed between the counting and rest conditions. In contrast to the short-latency components, P80 was significantly larger in the counting than in the rest condition, and showed a further increase from the counting to the movement condition. The N140 amplitude was significantly larger in the movement than the rest condition, but was not changed between the counting and the rest conditions. CONCLUSIONS: The attenuation of the frontal N30 and central N60, and the enhancement of the P80 and possibly the N140 resulted from the centrifugal mechanism. The present findings may show the different effects of voluntary movement on the early and subsequent cortical processing of the relevant somatosensory information requiring a behavioral response. SIGNIFICANCE: The present study demonstrated the differential modulation of short- and long-latency components of SEPs in a forewarned reaction time task. PMID- 15351363 TI - Impaired P50 sensory gating in Machado-Joseph disease. AB - OBJECTIVE: Machado-Joseph disease (MJD), an autosomal dominant spinocerebellar degeneration caused by an expanded CAG repeat on chromosome 14q32.1, is a disorder with wide range of neurological findings and brain regions involved. Studies evaluating neurophysiological parameters related to sensory gating in MJD are lacking. METHODS: This study intends to investigate P50 suppression, an auditory mid-latency evoked potential in a test-conditioning paradigm, considered as an index of sensory gating function. Twelve patients with MJD, 24 normal subjects and 12 schizophrenic patients were evaluated. RESULTS: MJD subjects had higher P50 ratios as compared to normal subjects (76.2 vs. 42.1%, P = 0.001), but similar to the group of schizophrenic patients. The difference from controls was due to greater test amplitudes (3.4 vs. 2.0 microV, P = 0.002), rather than to conditioning amplitudes. Latencies were higher for the MJD subjects than for controls (60.4 vs. 56.1 ms, P = 0.016). CONCLUSIONS: MJD may present sensory gating dysfunction. However, the pattern of this dysfunction seems to slightly differ from that classically found in schizophrenia, were both test and conditioning amplitudes seem to be implicated. SIGNIFICANCE: These results point out the P50 paradigm as a potential tool for further neurophysiological surveying in MJD. PMID- 15351364 TI - Influence of sleep stage and wakefulness on spectral EEG activity and heart rate variations around periodic leg movements. AB - OBJECTIVE: Typical changes in spectral electroencephalographic (EEG) activity and heart rate (HR) have been described in periodic leg movements (PLM) associated with or without microarousals (MA). We aimed to determine the effects of sleep stage and wakefulness on these responses to ascertain whether a common pattern of EEG and HR activation takes place. METHODS: The time course of EEG spectral activity and HR variability associated with PLM was analysed in 13 patients during light NREM sleep, rapid-eye-movement (REM) sleep and wakefulness. The same analysis was also conducted for PLM without MA occurring in stage 2. RESULTS: A significant EEG and electrocardiogram (ECG) activation was found associated with PLM during sleep, but not during wakefulness. While in light NREM sleep, an increase in delta and theta bands was detected before the PLM onset, in REM sleep the EEG activation occurred simultaneously with the PLM onset. Moreover, during stage 1 and REM sleep, alpha and fast frequencies tended to remain sustained after the PLM onset. In contrast, during wakefulness, a small and not significant increase in cerebral activity was present, starting at the PLM onset and persisting in the post-movement period. A typical pattern of cardiac response was present during NREM and REM sleep, the autonomic activation being lesser and prolonged during wakefulness. CONCLUSIONS: We conclude that the EEG and HR responses to PLM differ between sleep stages and wakefulness with lesser changes found during wakefulness. SIGNIFICANCE: These findings suggest that specific sleep state-dependent mechanisms may underlie the occurrence of PLM. PMID- 15351365 TI - Clinical correlates of periodic breathing in neonatal polysomnography. AB - OBJECTIVE: Periodic breathing is a respiratory pattern typical of preterm infants, but its clinical significance has not been clarified yet. The present study was designed to investigate whether the presence of periodic breathing is specifically associated to low post-conceptional ages, preterm birth, or common clinical disorders related to preterm birth. METHODS: The study included 271 consecutive infants submitted to neonatal polysomnography, of whom 138 were born before 37 complete gestational weeks (preterm) and 133 were full-term. The main outcome measure was periodic breathing. A multivariate analysis was performed to test the specific impact of preterm birth, respiratory distress syndrome and hypoxic-ischemic encephalopathy on the occurrence of periodic breathing, with adjustment for potential confounding factors such as the post-conceptional age by the time of the polysomnography. RESULTS: Periodic breathing was twice more frequent in infants born before term (83/138 [60%]) than in full-term babies (41/133 [31%], chi2 = 22.3, P < 0.0001). The presence of periodic breathing was not significantly associated to either hypoxic-ischemic encephalopathy or respiratory distress syndrome. After a multivariate analysis, only preterm birth remained in the regression model as a specific risk factor for periodic breathing (adjusted odds ratio=5.62, P < 0.0001). CONCLUSIONS: Periodic breathing is a respiratory pattern independently associated with preterm birth, and this association cannot be imputed to the coexistence of either respiratory distress syndrome or hypoxic-ischemic encephalopathy among preterm infants. SIGNIFICANCE: The finding of periodic breathing in neonatal polysomnography correlates with preterm birth but not with its associated clinical disorders. PMID- 15351366 TI - Age and gender effects in EEG coherence: I. Developmental trends in normal children. AB - OBJECTIVE: This study investigated intra-hemispheric and inter-hemispheric EEG coherences as a function of age and gender in normal children. METHODS: Two groups of 40 boys and 40 girls, each containing 8 normal children in each 1-year band from 8 to 12 years, participated. EEG was recorded from 21 sites during an eyes-closed resting condition. Wave-shape coherence was calculated for 8 intra hemispheric electrode pairs (4 in each hemisphere), and 8 inter-hemispheric electrode pairs, within each of the delta, theta, alpha and beta bands. RESULTS: There was consistent evidence of coherences developing across this age range, particularly in long-range intra- and inter-hemispheric electrode pairs. Coherences were larger in the left hemisphere than the right. Coherences also appeared to develop further in males than females, across several regions and frequency bands, suggesting that females lag males in this aspect of brain development. CONCLUSIONS: EEG coherences in normal children of this age range develop systematically with age. These developmental effects vary substantially with gender, brain region and frequency bands. SIGNIFICANCE: The data reported here provide basic norms for coherence development in different brain regions in normal children. PMID- 15351367 TI - Iron supplementation brings up a lacking P300 in iron deficient children. AB - OBJECTIVE: A decrease in iron concentration is accompanied by alterations in catecholaminergic and GABAergic neurotransmission systems, important in learning, memory and attention. It was hypothesized that iron deficient children would present attention deficits. A visual-event related potentials (ERPs) study is presented using an oddball paradigm in order to determine the P300 in ID children. METHODS: After medical examination, blood was obtained from 201 children for a complete hematological study. Two groups were selected, iron deficient (ID) (serum iron <60 microg/dl) and control (C) (serum iron >60 microg/dl). In both groups ERPs were recorded while executing a continuous performance task (oddball paradigm). Afterwards iron levels were restored in ID children by iron supplementation (ID-IS group) and all tests reapplied. RESULTS: ID children almost lacked a P300 in central and parietal regions. After iron supplementation, P300 clearly became evident although its Pz amplitude remained smaller compared to C children. CONCLUSIONS: A clear and strong correlation was found between ID and attention alterations in children. Iron supplementation nearly brings the P300 to normal levels although it is not known if the P300 difference in Pz is due to other nutritional/environmental deficits or to developmental psychomotor impairments in ID children. SIGNIFICANCE: It has been long known that iron deficient children have cognitive impairments but there is an insufficient number of electrophysiological works allowing to identify the source of this problem. In this work an attention deficit is demonstrated in ID children through a severely reduced P300, which recovers substantially after iron supplementation. PMID- 15351368 TI - The role of sensory and motor evoked potentials in the prognosis of Pott's paraplegia. AB - OBJECTIVE: In view of paucity of evoked potential changes in Pott's paraplegia, it is proposed to evaluate the role of motor and somatosensory evoked potentials in predicting the outcome. METHODS: Consecutive patients with Pott's paraplegia during 1993-2003 were subjected to detailed clinical, radiological and evoked potential study. The latter comprised of tibial somatosensory evoked potential (SEP) and motor evoked potential (MEP) study to tibialis anterior. The patients were clinically evaluated at 6 and 12 months and the outcome was defined into poor (bed ridden), partial (dependent for activities of daily living) and complete recovery (independent). The evoked potential findings were correlated with clinical and radiological findings and outcome. RESULTS: There were 39 patients whose age ranged between 16 and 70 (mean 42.1) years and 22 were females. The mean duration of symptoms was 8.2 months. Sensory motor deficit was present in 18 and pure motor signs in 21 patients. Five patients had quadriplegia and remaining had paraplegia. The muscle weakness was severe in 12 and moderate in 15 patients. In 12 patients, lower limb power was normal but they had lower limb hyper-reflexia with or without spasticity suggesting pyramidal dysfunction. Pinprick and joint position sensations were abnormal in 18 patients. MRI was abnormal in all and revealed cervical involvement in 7, thoracic in 22 and lumbar in 10 patients. Paravertebral soft tissue shadow was present in 36 and cord compression in 30 patients. Motor evoked potential was abnormal in 19 patients (unrecordable in 11 patients, 21 sides and prolonged in 8 patients, 14 sides). SEP was abnormal in 18 patients (unrecordable in 15 patients, 25 sides and prolonged central conduction in 8 patients, 9 sides). Both MEP and SEP were abnormal in 16, normal in 18, and only MEP was abnormal in 3 and only SEP in 2 patients. At 6 month 25 patients had complete, 9 partial and 5 poor recovery. At 1 year 33 had complete and 4 partial recovery. SEP and MEP abnormalities correlated with respective sensory and motor functions, vertebral level and outcome at 6 and 12 months. CONCLUSIONS: MEP and SEP both are helpful in predicting 6-month outcome. Combining SEP and MEP gives stronger correlation with 6-month outcome compared to only MEP or SEP. The potential role of evoked potentials in deciding different therapeutic strategies needs further studies. PMID- 15351369 TI - How large must an epileptic focus be to cause an electrographic status epilepticus--a case report. AB - Based on experimental data from animal studies different theories regarding the size of an epileptic focus have been postulated which range from single pacemaker cells to extended neuronal networks. We report a case which gives further information about the size of a human epileptic focus which can trigger manifest epileptic seizures. We report a 22-year-old man with medically refractory temporal lobe epilepsy. This patient suffered from brief complex partial seizures and frequent epigastric auras. To differentiate a mesiotemporal from a temporolateral seizure origin the patient was implanted with a 10 contact depth electrode from a posterior approach into the right hippocampus, and additional temporobasal/temporolateral subdural strip electrodes. Depth recordings revealed an electrographic status with continuous rhythmic sharp wave activity (1 Hz), the field of which was confined to a diameter of less than 1 cm in the anterior hippocampus, whereas temporobasal subdural strip electrodes did not display this activity. Periodically, spread of this activity occurred to the amygdala, to the posterior part of the hippocampus, and less often to the temporobasal cortex. Most seizure patterns remained subclinical, few of them became symptomatic as partial seizures. This case demonstrates that a hippocampal epileptic focus causing electrographic focal status epilepticus may be limited to a volume of less than 1 cm in diameter. This observation is discussed with regard to implantation strategies and to possible superselective resective or modulatory approaches in the treatment of such limited epileptogenic areas. PMID- 15351370 TI - Seizure detection: evaluation of the Reveal algorithm. AB - OBJECTIVE: The aim of this study is to evaluate an improved seizure detection algorithm and to compare with two other algorithms and human experts. METHODS: 672 seizures from 426 epilepsy patients were examined with the (new) Reveal algorithm which utilizes 3 methods, novel in their application to seizure detection: Matching Pursuit, small neural network-rules and a new connected object hierarchical clustering algorithm. RESULTS: Reveal had a sensitivity of 76% with a false positive rate of 0.11/h. Two other algorithms (Sensa and CNet) were tested and had sensitivities of 35.4 and 48.2% and false positive rates of 0.11/h and 0.75/h, respectively. CONCLUSIONS: This study validates the Reveal algorithm, and shows it to compare favorably with other methods. SIGNIFICANCE: Improved seizure detection can improve patient care in both the epilepsy monitoring unit and the intensive care unit. PMID- 15351371 TI - Identifying true brain interaction from EEG data using the imaginary part of coherency. AB - OBJECTIVE: The main obstacle in interpreting EEG/MEG data in terms of brain connectivity is the fact that because of volume conduction, the activity of a single brain source can be observed in many channels. Here, we present an approach which is insensitive to false connectivity arising from volume conduction. METHODS: We show that the (complex) coherency of non-interacting sources is necessarily real and, hence, the imaginary part of coherency provides an excellent candidate to study brain interactions. Although the usual magnitude and phase of coherency contain the same information as the real and imaginary parts, we argue that the Cartesian representation is far superior for studying brain interactions. The method is demonstrated for EEG measurements of voluntary finger movement. RESULTS: We found: (a) from 5 s before to movement onset a relatively weak interaction around 20 Hz between left and right motor areas where the contralateral side leads the ipsilateral side; and (b) approximately 2-4 s after movement, a stronger interaction also at 20 Hz in the opposite direction. CONCLUSIONS: It is possible to reliably detect brain interaction during movement from EEG data. SIGNIFICANCE: The method allows unambiguous detection of brain interaction from rhythmic EEG/MEG data. PMID- 15351372 TI - On the spatio-temporal organisation of quadratic phase-couplings in 'trace alternant' EEG pattern in full-term newborns. AB - OBJECTIVE: The time courses of quadratic phase-coupling (QPC) of electroencephalographic burst and interburst patterns of the 'trace alternant' (TA) in full-term newborns have been quantified. METHODS: Using the Gabor expansion, a fast Fourier transformation based method, biamplitude, bicoherence and phase-bicoherence time courses of both burst and interburst patterns have been determined (common average reference EEG recordings). With a frequency resolution of 0.25 Hz and a frequency grid of 1-1.5 <==> 3.5-4.5 Hz (region-of interest), a number of 15 frequency pairs result. These pairs have been investigated. RESULTS: The burst and the interburst patterns are characterized by temporally and topographically different QPC profiles. All differences are dominant at the electrode Fp1 followed by Fp2. There is a significant difference (combined multiple and global test strategy) in the QPC characteristics between both patterns within the time period from 0.75 to 1.5 s after the pattern onset at electrode Fp1. The maximal QPC in burst patterns (especially at Fp1) can be observed during this time period. In contrast to this finding, maximal QPC in interburst patterns (at Fp1) are reached immediately after the onset and at 3 s. Summarising all findings, a QPC-rhythm of 0.1 Hz during TA can be assumed. CONCLUSIONS: It can be assumed that the QPC rhythm of the TA is generated by a pattern-spanning time-variant phase-locking process and there are indications for a possible correspondence between the QPC rhythm and vegetative rhythms. SIGNIFICANCE: This study showed that advanced, time-variant analysis methods quantifying QPC rhythms are able to add new scientific information to the understanding of nature, characteristics and significance of TA in the neonatal EEG. PMID- 15351373 TI - Medial antebrachial cutaneous nerve conduction study, a new tool to demonstrate mild lower brachial plexus lesions. A report of 16 cases. AB - OBJECTIVE: This was to demonstrate the ability to electrodiagnosed mild lower brachial plexus lesion only through abnormal medial antebrachial cutaneous nerve (MABCN) conduction study. METHODS: We report 16 cases of unilateral, atypical pains and paresthesias of the upper limbs without motor deficit or atrophy. Patients were referred as carpal tunnel syndrome in 12 cases. All patients had needle examination of the impaired upper limb from C5 to T1. Motor and sensory conductions of median and ulnar nerves were bilaterally studied. MABCN was antidromically (16 cases) and orthodromically (9 cases) studied at the elbow in the both sides. MABCN abnormality was defined by an interside amplitude ratio of the sensory nerve action potential equal or greater than 2 (mean + 3 SD). RESULTS: No patient had a definitive and accurate diagnosis, before MABCN abnormality determination. MABCN testing was abnormal in all the 16 cases with a mean interside amplitude ratio of 7.2 (mean + 25 SD), when all other motor and sensory nerve conductions were normal. All except four patients showed normal needle examination from C5 to T1. In 5 cases, an obvious cause (traumatic and neoplastic) explained the mild lower brachial plexus lesion. In 2 cases, a mild neurogenic thoracic outlet syndrome (NTOS) was confirmed by surgical findings. In the 9 other cases, the mild lower brachial plexus lesion defined by MABCN findings, was without cause and was considered as a mild NTOS. CONCLUSIONS AND SIGNIFICANCE: These 16 cases, support a new electrodiagnostic pattern to define a mild lower brachial plexus lesion: comparatively low or low MABCN SNAP amplitude, normal median and ulnar SNAP/Compound motor action potential amplitudes and normal or slightly reduced interference pattern in some C8-T1 innervated muscles. This pattern can be found in patients with 'carpal tunnel syndrome like' symptoms who have normal electro-diagnostic examination, or in patients with clinical features suggesting a lower brachial plexus lesion. PMID- 15351374 TI - Pure motor mononeuropathy with distal conduction block: an unusual presentation of multifocal motor neuropathy with conduction blocks. AB - OBJECTIVE: To report an hitherto undescribed presentation of Motor Neuropathy with Multifocal Conduction Block (MMNCB). METHODS: Description of two patients presenting with complete paralysis of the 3 heads of the trapezius muscle (case one) and progressive weakness of finger extension (case 2). RESULTS: Nerve conduction studies (NCS) established that the corresponding nerves were distally inexcitable. In the affected muscles, no voluntary activity was elicited in both patients with spontaneous activity noted in patient 2. Systematic NCSs documented an asymptomatic, partial conduction block (CB) in a median nerve forearm segment in both patients. Neurophysiological follow-up after a dramatic response to intravenous immunoglobulins demonstrated recovery of the initially unobtainable motor responses. CONCLUSIONS: This indicates that a complete, distal CB of the motor fibers destined to the trapezius muscle in patient 1, and to the extensor indicis proprius in patient 2, had caused the heralding deficits. SIGNIFICANCE: These findings underscore the possibility of distal CB in this disorder and the need for extensive NCSs, including asymptomatic nerves, for an accurate diagnosis. PMID- 15351375 TI - Neurophysiologic findings in early acute inflammatory demyelinating polyradiculoneuropathy. AB - BACKGROUND: Patients with early acute inflammatory demyelinating polyradiculoneuropathy (AIDP) may not meet the current neurophysiologic criteria. OBJECTIVE: To document neurophysiologic findings in early AIDP. METHODS: Clinical and neurophysiologic data from 38 AIDP patients, assessed within 10 days of symptom onset were reviewed. RESULTS: In addition to absent H reflexes and abnormal F-wave responses in the acute stage of AIDP, abnormalities of blink reflexes, upper limb sensory responses abnormalities with spared sural responses, presence of distal CMAP dispersion, and A-waves were frequently observed. Established demyelinating neurophysiologic parameters were infrequently seen. CONCLUSIONS: Abnormalities of H reflexes and F responses were most frequently noted in early AIDP. Additionally, distal temporal dispersion, prolonged or absent blink reflexes, and A-waves were often present in the acute stage of AIDP when classic diagnostic criteria of AIDP were not satisfied. SIGNIFICANCE: Neurophysiologic studies in early AIDP frequently reveal abnormalities that are not specific of primary demyelinating neuropathy. PMID- 15351376 TI - Various aspects of F-wave values in a healthy population. AB - OBJECTIVE: To characterize various aspects of F-wave in a healthy population and establish normative data for future clinical use. METHODS: A total of 100 healthy volunteers underwent sensory and motor nerve conduction studies of the ulnar and tibial nerves, including F waves elicited by 32 stimuli. RESULTS: The F-wave measurements (mean +/- SD for ulnar vs tibial nerve) consisted of persistence (83 +/- 19 vs 97 +/- 5%), minimum, mean and maximum latencies (26.5 +/- 2.1, 28.1 +/- 2.2, and 30.4 +/- 2.3 vs 47.0 +/- 4.1, 49.6 +/- 4.4, and 52.5 +/- 4.4 ms), minimum, mean and maximum F-wave conduction velocities (FWCV) (55.0 +/- 2.7, 60.0 +/- 2.3, and 64.0 +/- 3.0 vs 49.0 +/- 2.9, 52.2 +/- 3.1, and 55.5 +/- 3.4 m/s), chronodispersion (3.9 +/- 0.9 vs 5.5 +/- 1.4 ms), mean amplitude (347 +/- 152 vs 384 +/- 148 microV) and mean duration (8.6 +/- 2.9 vs 13.0 +/- 4.5 ms). Additional measures, registered by electronic averaging, included latency (27.4 +/- 2.3 vs 48.6 +/- 4.7 ms), duration (9.6 +/- 2.2 vs 16.4 +/- 4.2 ms), and amplitude (299 +/- 156 vs 208 +/- 116 microV). CONCLUSIONS: The use of a height nomogram serves well as an acceptable means to adjust F latencies for the limb length. In addition to the commonly used minimal latency, maximal FWCV, and persistence, clinically relevant measures with a narrow variability includes mean and maximal latencies, chronodispersion, and mean duration. In particular, mean latency obtained with 10 stimuli gave accurate results either for group or individual analysis. SIGNIFICANCE: The data help establish an adequate manner of recording F-wave latencies in clinical evaluation. PMID- 15351377 TI - Changes in the perioral muscle responses to cortical TMS induced by decrease of sensory input and electrical stimulation to lower facial region. AB - OBJECTIVE: To determine the changes in the motor cortex due to repetitive electrical stimulation and cutaneous anesthesia in lower facial region. METHODS: A total of 11 subjects participated in the study of repetitive electrical stimulation, and 10 other subjects in the study of lower facial anesthesia. Facial nerve root and face associated cortical MEPs by transcranial magnetic stimulation (eight-shaped coil) were recorded from perioral muscles pre- and post electrical stimulation and lower facial anesthesia. Cheek near to the corner of the mouth was transcutaneously stimulated by bipolar surface electrode giving repetitive electrical shocks at 5 Hz. Five percent lidocain/prilocain local anesthetic cream was applied to left or right lip-cheek region. RESULTS: There was no significant change in perioral MEP responses after 10-30 min of 5 Hz electrical stimulation. We found a significant increase of amplitude in cortical MEP recordings during lower facial anesthesia especially in cases of cortical magnetic stimulations ipsilateral and contralateral to the anaesthetized side and in perioral recordings contralateral to the anaesthetized side. CONCLUSIONS: The present study demonstrates that topical anesthesia to the lower facial region leads to cortical modulation and fast plastic changes in both hemispheres that are directed to the normal side. PMID- 15351378 TI - Tendon reflexes for predicting movement recovery after acute spinal cord injury in humans. AB - OBJECTIVE: Use the tendon reflex to examine spinal cord excitability after acute spinal cord injury (SCI), relating excitability findings to prognosis. METHODS: We conducted repeated measures of reflex responses to mechanical taps at the patellar and Achilles tendons of the lower limbs, and the wrist flexor tendons of the upper limbs in persons with acute SCI, beginning as early as the day of injury. The single largest EMG response (peak-to-peak) for each site was recorded. Subjects were compared based on level of injury and final neurologic status of lower limb motor function (i.e. absence of any voluntary recruitment in a lower limb muscle: motor-complete; voluntary recruitment in 1 or more lower limb muscles: motor-incomplete). RESULTS: We studied 229 subjects with acute SCI. Persons with injury to the cervical or thoracic spinal cord and who were (or became) motor-incomplete showed large tendon responses, even at the time of initial evaluation. In combination with larger tendon response amplitudes, the presence of the 'crossed-adductor' response to patellar tendon taps at the acute stage was highly predictive of functional motor recovery following SCI. In marked contrast, tendon responses were small (e.g. < 0.1 mV) or absent in persons with acute, motor-complete injury (and which remained motor-complete), and the crossed adductor response was never seen. Reflex amplitudes and the incidence of the crossed-adductor response increased somewhat over time in persons with motor complete SCI, but did not approach the values seen in motor-incomplete subjects. CONCLUSIONS: Taken together, tendon response amplitude and reflex spread were sensitive and specific indicators of preserved supraspinal control over lower limb musculature in subjects with acute SCI. A simple algorithm using these outcome measures predicted a 'motor-complete' status with 100% accuracy, and a motor-incomplete status with accuracy exceeding 91%. PMID- 15351379 TI - Physiological measures of therapeutic response to interferon beta-1a treatment in remitting-relapsing MS. AB - OBJECTIVE: This pilot study was designed to determine the effects of interferon beta-1a (IFNB) therapy (Avonex) on cortically evoked motor potentials (MEPs) during resting and fatigued states in individuals with multiple sclerosis (MS). METHODS: Eight women with relapsing-remitting MS (mean age 36) and mean Expanded Disability Status Scale (EDSS) score of 3.1 were evaluated before and after 3, 6, and 12 months of IFNB therapy. At each test period, MEPs were recorded at rest and following a fatigue paradigm (3 min maximal contraction). Effects of IFNB on neurological and functional (7.7 m walk and 10 s finger tapping) status and fatigue were also examined. RESULTS: Recovery from post-exercise depression of MEP amplitudes (PED) was 41, 43, and 43.5% faster at 3, 6, and 12 months, respectively, compared to baseline (P < 0.05). Percent reduction of MEP amplitude was significantly less at 6 months (P < 0.05) The majority of subjects (5/8 at 3 months; 6/8 at 6 and 12 months) reported decreased physical fatigue. Functional improvements were observed for walk and finger tapping scores after 3 months of IFNB treatment. MEP latencies were unchanged over the course of the intervention. CONCLUSIONS: Results indicate that IFNB therapy may improve the rate of recovery from central fatigue. SIGNIFICANCE: Transcranial magnetic stimulation (TMS) may have promise as an objective physiological tool to evaluate disease activity and treatment responses in MS. PMID- 15351380 TI - Altered central nervous system signal during motor performance in chronic fatigue syndrome. AB - OBJECTIVE: The purpose of this study was to determine whether brain activity of chronic fatigue syndrome (CFS) patients during voluntary motor actions differs from that of healthy individuals. METHODS: Eight CFS patients and 8 age- and gender-matched healthy volunteers performed isometric handgrip contractions at 50% maximal voluntary contraction level. They first performed 50 contractions with a 10 s rest between adjacent trials--'Non-Fatigue' (NFT) task. Subsequently, the same number of contractions was performed with only a 5 s rest between trials -'Fatigue' (FT) task. Fifty-eight channels of surface EEG were recorded simultaneously from the scalp. Spectrum analysis was performed to estimate power of EEG frequency in different tasks. Motor activity-related cortical potential (MRCP) was derived by triggered averaging of EEG signals associated with the muscle contractions. RESULTS: Major findings include: (i) Motor performance of the CFS patients was poorer than the controls. (ii) Relative power of EEG theta frequency band (4-8 Hz) during performing the NFT and FT tasks was significantly greater in the CFS than control group (P < 0.05). (iii) The amplitude of MRCP negative potential (NP) for the combined NFT and FT tasks was higher in the CFS than control group (P < 0.05) (iv) Within the CFS group, the NP was greater for the FT than NFT task (P<0.01), whereas no such difference between the two tasks was found in the control group. CONCLUSIONS: These results clearly show that CFS involves altered central nervous system signals in controlling voluntary muscle activities, especially when the activities induce fatigue. SIGNIFICANCE: Physical activity-induced EEG signal changes may serve as physiological markers for more objective diagnosis of CFS. PMID- 15351381 TI - Characterising the central mechanisms of sensory modulation in human swallowing motor cortex. AB - OBJECTIVE: Pharyngeal stimulation can induce remarkable increases in the excitability of swallowing motor cortex, which is associated with short-term improvements in swallowing behaviour in dysphagic stroke patients. However, the mechanism by which this input induces cortical change remains unclear. Our aims were to explore the stimulus-induced facilitation of the cortico-bulbar projections to swallowing musculature and examine how input from the pharynx interacts with swallowing motor cortex. METHODS: In 8 healthy subjects, a transcranial magnetic stimulation (TMS) paired-pulse investigation was performed comprising a single conditioning electrical pharyngeal stimulus (pulse width 0.2 ms, 240 V) followed by cortical TMS at inter-stimulus intervals (ISI) of 10-100 ms. Pharyngeal sensory evoked potentials (PSEP) were also measured over the vertex. In 6 subjects whole-brain magnetoencephalography (MEG) was further acquired following pharyngeal stimulation. RESULTS: TMS evoked pharyngeal motor evoked potentials were facilitated by the pharyngeal stimulus at ISI between 50 and 80 ms (Delta mean increase: 47+/-6%, P < 0.05). This correlated with the peak latency of the P1 component of the PSEP (mean 79.6+/-8.5 ms). MEG confirmed that the equivalent P1 peak activities were localised to caudolateral sensory and motor cortices (BA 4, 1, 2). CONCLUSIONS: Facilitation of the cortico-bulbar pathway to pharyngeal stimulation relates to coincident afferent input to sensorimotor cortex. SIGNIFICANCE: These findings have mechanistic importance on how pharyngeal stimulation may increase motor excitability and provide guidance on temporal windows for future manipulations of swallowing motor cortex. PMID- 15351382 TI - Perceived effort in force production as reflected in motor-related cortical potentials. AB - OBJECTIVE: The perceived effort in force production was investigated in a series of experiments where subjects performed isometric force tasks with the index finger while the nominal force, the rate of force development and signal gain were controlled and rating of effort for each task was obtained. The hypotheses tested were that: (1) force-related perceived effort may selectively influence the amplitude of motor-related cortical potentials (MRCP); and (2) the MRCP may directly reflect the intensity of perceived effort associated with force production. METHODS: The force trace was displayed on a computer monitor using various control-gains so that the perceived effort matched or was at odds with actual muscular effort applied to the load cell to accomplish the task. The MRCP were extracted from continuous EEG records using averaging techniques. RESULTS: The findings showed that: (1) perceived effort proportionally increased with the increment of rate of force development and force error, but not with the actual force level; (2) the amplitude of the MRCP increased when a large amount of force was accompanied by an increased rate of force development; (3) the amplitude of early components of MRCP preceding the force initiation (MP-100 to 0) increased as a function of anticipated effort, whereas, the amplitude movement-monitoring potentials (MMP) accompanying the force production increased as a function of actual force level. CONCLUSIONS: The findings from this study provide additional insight clarifying the distinct patterns of EEG activity exhibited under various degrees of perceived effort associated with force output. The findings support the hypothesis that the early components of MRCP may reflect the perceived effort associated with achieving the required force level. SIGNIFICANCE: The results from this study may be considered in the larger context of physical activity in terms of importance of perceived effort during prescribed exercise in rehabilitation programs. PMID- 15351383 TI - Sparing of perceptual implicit memory despite suppression of the electrophysiological word repetition effect: a study with general anesthetic isoflurane. AB - OBJECTIVE: Much effort has been devoted to the search for the neurophysiological correlates of implicit memory. A commonly held view is that the early portion (250-500 ms) of the event-related potential (ERP) word repetition effect reflects processes important for perceptual implicit memory whereas the latter portion reflects processes implicated in explicit memory. It is, however, difficult to disentangle with certainty the relative contributions of each form of memory on ERPs since both forms co-exist in normal subjects. To dissociate ERP effect related to implicit and explicit memory, we used isoflurane sedation in normal subjects to suppress explicit remembering while sparing implicit memory. These ERPs were compared with those of non-medicated control subjects. METHODS: Thirteen subjects performed an incidental encoding task for words presented auditorily during the inhalation of a subanesthetic dose of isoflurane. After termination of isoflurane administration, we assessed free recall and recorded ERPs during a syllable completion task (implicit memory) and during a passive listening task (ERP repetition effect). Eleven non-medicated control subjects were tested in a similar manner. RESULTS: The controls showed robust early and late ERP repetition effect. The isoflurane group had implicit memory without free recall and showed no ERP repetition effect. CONCLUSIONS: These findings failed to show an association between any part of the repetition effect and perceptual implicit memory. The results are consistent with the view that processes linked to explicit memory contribute to the ERP repetition effect since there was a marked difference in free recall between the control and isoflurane groups. SIGNIFICANCE: The present study shows that the reversible alterations of memory by general anesthetics can be used to study the neurophysiological correlates of memory processes. PMID- 15351384 TI - Different TMS patterns of intracortical inhibition in early onset Alzheimer dementia and frontotemporal dementia. AB - OBJECTIVE: To investigate putative changes in cortical excitability of patients affected by early-onset mild dementia by means of transcranial magnetic stimulation (TMS) and to verify whether a peculiar neurophysiological profile may contribute to characterise Alzheimer's disease (AD) vs frontotemporal dementia (FTD). METHODS: Motor threshold and intracortical inhibition (ICI) and facilitation (ICF) after paired-pulse TMS (inter-stimulus intervals from 1 to 20 ms) were studied in two groups of early-onset demented patients with a neuropsychological profile suggestive of AD (n = 12) and FTD (n = 8). Twelve age matched healthy subjects were considered as control group. In both patient groups, recordings were performed before and after a single oral dose of 4 mg galantamine. RESULTS: No significant difference in motor threshold was observed among the three studied groups. On the contrary, early-onset AD showed a significant reduction of ICI compared to control group, no changes were detected in FTD patients. No significant changes in ICF were found between both patient groups and healthy subjects. The acute administration of galantamine reversed the modified ICI in AD group. CONCLUSIONS: The differential pattern of ICI exhibited by early-onset AD vs FTD in the early stage of disease may represent a non invasive, reproducible electrophysiological tool, which may contribute to early differential diagnosis and, possibly, to monitor therapeutic effectiveness. SIGNIFICANCE: The present results support the possibility that subtle, early modifications in intracortical circuitry features AD, but not FTD patients. PMID- 15351385 TI - MRI study of human brain exposed to weak direct current stimulation of the frontal cortex. AB - OBJECTIVE: To determine whether weak transcranial direct current stimulation (tDCS), which is an interesting new tool inducing prolonged cortical excitability shifts in humans, induces brain edema, disturbance of the blood-brain barrier or structural alterations of the brain detectable by magnetic resonance imaging (MRI). METHODS: In 10 healthy individuals, tDCS, which is known to alter cortical excitability for about 1 h, was applied over motor and pre-frontal cortices. contrast-enhanced t1-, t2-, and diffusion-weighted mri was performed immediately before, 30 and 60 min after tdcs. RESULTS: MRI performed 30 and 60 min after tDCS did not show pathological signal alterations in pre- and post-contrast-enhanced T1-weighted and diffusion-weighted MR sequences. CONCLUSIONS: tDCS protocols which are known to result in cortical excitability changes persisting for an hour after stimulation do not induce brain edema or alterations of the blood-brain barrier or cerebral tissue detectable by MRI. SIGNIFICANCE: These results deliver further evidence for the safety of the currently applied tDCS protocols in humans. PMID- 15351386 TI - Cocaine pharmacology and current pharmacotherapies for its abuse. AB - Cocaine abuse continues to be prevalent and effective therapies for cocaine craving and addiction remain elusive. In the last decade immunopharmacotherapy has been proposed as a promising means to alleviate this illness. By using the organism's natural immune response, an anti-cocaine vaccine promotes the production of cocaine-specific antibodies that sequester the drug before their passage into the brain, where it exerts its reinforcing and thus addictive effects. A series of studies demonstrating the cocaine-blocking properties of various immunogenic conjugates will be reviewed in the context of the neuropsychopharmacological profile of the drug. PMID- 15351387 TI - Anchimeric assistance effect on regioselective hydrolysis of branched PEGs: a mechanistic investigation. AB - Branched poly(ethylene glycols) (PEG2) are nowadays widely used for protein and peptides bioconjugation, for their favourable properties (such as the ability to protect the protein surface in an 'umbrella like' fashion). The discovery that mPEG(2)-LysMetbeta AlaOEt lost one mPEG chain during standard base-catalysed ester hydrolysis conditions prompted us to investigate the hydrolytic stability of such systems and the mechanism involved in the PEG chain loss. A series of branched PEGs, substituted with different aminoacids and dipeptides, have been prepared to test the influence of steric hindrance, chain lengths, ramification and Lys-AA amide substitution on hydrolysis. Unexpected results reveal an anchimeric assistance of the Lys-AaA amide proton to the hydrolysis of the carbamoyl moiety joining mPEG to the alpha-amino group of lysine through the formation of an hydantoin system. PMID- 15351388 TI - Synthesis and structure-activity relationships of phenoxypyridine derivatives as novel inhibitors of the sodium-calcium exchanger. AB - The sodium-calcium exchanger (NCX) is known as the transporter that controls the concentration of Ca(2+) in cardiac myocytes. In the setting of heart failure and myocardial ischemia-reperfusion, NCX underlies an arrhythmogenic transient inward current responsible for delayed after--depolarizations and nonreentrant initiation of ventricular tachycardia. NCX is an attractive target for treatment in heart failure and myocardial ischemia-reperfusion. We have designed and synthesized a series of phenoxypyridine derivatives, based on compound 3. These derivatives have been evaluated for their inhibitory activity against both the reverse and forward mode of NCX in CCL39 cells. We have discovered several novel potent NCX inhibitors (39q, 48k), which have a high selectivity for reverse NCX inhibitory activity. PMID- 15351389 TI - Designed calix[8]arene-based ligands for selective tryptase surface recognition. AB - Basic amino acid calix[8]arene receptors for tryptase surface recognition have been synthesized. The tetrameric arrangement and the negative charge distribution close to the active sites of the enzyme, have suggested the design of complementary multifunctional receptors that might bind to the active region of the protein blocking the approach of the substrate. Kinetic inhibition analysis on recombinant lung tryptase have showed a time-dependent competitive inhibition with both initial and steady-state rate constants in the nanomolar range. PMID- 15351390 TI - Design, synthesis, and biological evaluation of new phosphodiesterase type 4 inhibitors. AB - The design, synthesis, and biological evaluation of new phosphodiesterase type 4 inhibitors, which possess new templates instead of a cyclohexane ring, are described. The mode of interaction with the enzyme is discussed based on the structure-activity relationship (SAR) data obtained for the synthesized inhibitors. Furthermore, the roles of three pharmacophores, a catechol moiety, a nitrile moiety, and acidic moieties, are discussed using in silico docking studies. More detailed biological evaluations of selected compounds are also presented. PMID- 15351391 TI - Synthesis and evaluation of thymine-derived carboxamides against mitochondrial thymidine kinase (TK-2) and related enzymes. AB - Based on the structure of our previously identified mitochondrial thymidine kinase (TK-2) inhibitors, three series of thymine-derived carboxamides have been synthesized and tested against TK-2 and related enzymes. The methodology employed has been a solution-phase parallel synthesis based on the coupling of three thymine-derived acids [4-(thymin-1-yl)butyric acid (I), [4-(thymin-1-yl) butyrylamino]acetic acid (II) and 6-(thymin-1-yl)hexanoic acid (III)] with different commercially available primary amines that carry cyano and/or phenyl groups. The couplings were performed in good yields (from 60% to 90%), with the exception of those that incorporate the highly crowded triphenylmethylamine (e). From the new synthesized compounds, the N-trityl-6-(thymin-1-yl)hexanamide (IIIe) was the most active TK-2 inhibitor (IC(50)=19+/-2microM). PMID- 15351392 TI - Synthesis of polyhydroxylated piperidines and evaluation as glycosidase inhibitors. AB - A series of 16 new chiral nonracemic polyhydroxylated piperidines was synthesized utilizing several chiral beta-amino-alcohols. They act as a nitrogen source, chirality inducer and iminium stabilizer, in the desymmetrization of meso trihydroxylated glutaraldehyde. The biological activity of these compounds towards several glycosidases (alpha-D-glucosidase, alpha-D-mannosidase, alpha-L fucosidase) has been evaluated. PMID- 15351393 TI - Synthesis and biological activity of N-terminal lipidated and/or fluorescently labeled conjugates of astressin as corticotropin releasing factor antagonists. AB - This report describes the synthesis of eight N-terminally modified astressin analogs and their biochemical evaluation as corticotropin releasing factor (CRF) antagonists. The lipidated astressin derivatives were tested on rat CRF receptor type 1 and 2alpha and were found to be active as CRF antagonists (rCRFR1: pA(2)=7.5-8.3; rCRFR2alpha: pA(2)=7.5-9.0) with nearly equal activities as compared to unmodified astressin (rCRFR1: pA(2)=8.3+/-0.09; rCRFR2alpha: pA(2)=8.7+/-0.08). PMID- 15351394 TI - Synthesis, antimicrobial, and anti-HIV-1 activity of certain 5-(1-adamantyl)-2 substituted thio-1,3,4-oxadiazoles and 5-(1-adamantyl)-3-substituted aminomethyl 1,3,4-oxadiazoline-2-thiones. AB - The reaction of 5-(1-adamantyl)-1,3,4-oxadiazoline-2-thione 2 with iodoethane, 2 dimethylaminoethyl chloride hydrochloride or 2-piperidinoethyl chloride hydrochloride in ethanolic potassium hydroxide yielded the corresponding 5-(1 adamantyl)-2-ethyl or substituted ethylthio-1,3,4-oxadiazoles 3a-c. Interaction of 2 with formaldehyde solution and primary aromatic amines or 1-substituted piperazines, in ethanol at room temperature yielded the corresponding 5-(1 adamantyl)-3-arylaminomethyl-1,3,4-oxadiazoline-2-thiones 4a-m or 5-(1-adamantyl) 3-(4-substituted-1-piperazinylmethyl)-1,3,4-oxadiazoline-2-thiones 5a-h, respectively. All the synthesized compounds were tested for in vitro activities against certain strains of Gram-positive and Gram-negative bacteria and the yeast like pathogenic fungus Candida albicans. Compounds 2, 5a, and 5e were found as the most active derivatives, particularly against the Gram-positive bacteria. In addition, the antiviral activity of compounds 2, 4a-m, and 5a-h against HIV-1 using the XTT assay was carried out. Compound 2 produced 100%, 43%, and 37% reduction of viral replication at 50, 10, and 2microg/mL concentrations, respectively. PMID- 15351395 TI - Combinatorial synthesis of substituted 3-(2-indolyl)piperidines and 2-phenyl indoles as inhibitors of ZipA-FtsZ interaction. AB - The ZipA-FtsZ protein-protein interaction is a potential target for antibacterial therapy. The design and parallel synthesis of a combinatorial library of small molecules, which target the FtsZ binding area on ZipA are described. Compounds were demonstrated to bind to the FtsZ binding domain of ZipA by HSQC NMR and to inhibit cell division in a cell elongation assay. PMID- 15351396 TI - Evaluation of chelating agents as anti-angiogenic therapy through copper chelation. AB - The evaluation of several sets of polyamine donor chelating agents including a selection of novel hexadentate 1,3,5-cis,cis-triaminocyclohexane (tach) based derivatives were performed in an in vitro endothelial cell proliferation assay to assess their cytotoxicity and selectivity as novel anti-angiogenic agents. The selective nature of the anti-angiogenic agents for human umbilical vein endothelial cells (HUVEC) was compared to a normal fibroblast cell line and a human Glioma cell line to evaluate these compounds. Linear tri- and tetra polyamines were superior to both macrocyclic and the tach based polyamine chelating agents in terms of selectivity of its inhibitory activity toward the proliferation of HUVEC cells compared to the fibroblast and human Glioma cells. The linear polyamine, triethylenetetramine (22), previously reported to possess anti-angiogenic properties failed to demonstrate any selectivity for inhibiting the proliferation of HUVEC cells compared to the fibroblast and human Glioma cells. PMID- 15351397 TI - Antioxidant activities of flavidin in different in vitro model systems. AB - Flavidin was isolated from Orchidaceae species and purified by silica gel column chromatography. The structure was identified using physical and spectral ((1)H, (13)C NMR, and mass) data. Antioxidant potency of flavidin was investigated employing various established in vitro model systems viz., beta-carotene linoleate, 1,1-diphenyl-2-picryl hydrazyl (DPPH), phosphomolybdenum method, and scavenging of hydrogen peroxide methods. Flavidin showed very good antioxidant activity (90.2%) and almost equivalent to that of BHA at 50ppm level by beta carotene-linoleate method. Radical scavenging activity of flavidin was compared with BHA at 5, 10, 20, and 40ppm concentration and flavidin showed more radical scavenging activity than BHA at all the tested concentrations. Furthermore, flavidin showed very good antioxidant capacity by the formation of phosphomolybdenum complex method. Besides this, flavidin showed effective hydrogen peroxide scavenging activity. The data obtained in the in vitro models clearly establish the antioxidant potency of flavidin. However, comprehensive studies need to be conducted to ascertain the in vivo safety of flavidin in experimental animal models. This is the first report on antioxidant activity of 9,10-dihydro-5H-phenanthro-(4,5 bcd)-pyrans/flavidin type of compounds. PMID- 15351398 TI - Design, synthesis, and biological evaluation of cytotoxic 11 aminoalkenylindenoisoquinoline and 11-diaminoalkenylindenoisoquinoline topoisomerase I inhibitors. AB - The cytotoxic indenoisoquinolines are a novel class of noncamptothecin topoisomerase I inhibitors having certain features that compare favorably with the camptothecins. A new strategy was adopted to attach aminoalkenyl substituents at C-11 of the indenoisoquinoline ring system, which, according to molecular modeling, would orient the side chains toward the DNA minor groove. All of the newly synthesized compounds were more cytotoxic than the parent indenoisoquinoline NSC 314622. Despite an imperfect correlation between cytotoxicities and topoisomerase I inhibition results, the hypothetical structural model of the cleavage complex presented here provides a conceptual framework to explain the structure-activity relationships. PMID- 15351399 TI - The anandamide membrane transporter. Structure-activity relationships of anandamide and oleoylethanolamine analogs with phenyl rings in the polar head group region. AB - A new series of anandamide and N-oleoylethanolamine analogs, most of which with aromatic moieties in the head group region, has been synthesized and evaluated as inhibitors of anandamide uptake. Some of them efficaciously inhibit the uptake process with K(i) values in the low micromolar range (2.4-21.2microM). Strict structural requisites are needed to observe a significant inhibition and in no case inhibition of fatty acid amidohydrolase overlaps with inhibition of anandamide uptake. PMID- 15351400 TI - Design, synthesis, and activity of caffeoyl pyrrolidine derivatives as potential gelatinase inhibitors. AB - The synthesis and biological evaluation of caffonyl pyrrolidine derivatives as MMPs inhibitors are reported in this paper. Inhibiting activities of synthesized compounds on gelatinase (MMP-2 and -9) were tested by using succinylated gelatin as substrate. Structure-activity relationship results from these tested compounds demonstrated that longer and more flexible side chain linked to the pyrrolidine ring at C(4) produced higher activity at gelatinase. Furthermore, aromatic heterocycle and sulfamide in the same position could enhance the activities. Compounds with free phenol hydroxyl group showed higher activity compared to methylated derivatives (or counterparts), which confirms the importance of phenol hydroxyl functionality in the interaction with gelatinase. The anti-metastasis model of mice bearing H(22) tumor cell was used to evaluate their in vivo inhibiting activities. All tested compounds were orally administered at a dose of 50 or 100mg/kg, 6days/week for two weeks. The test results demonstrated that most of these inhibitors showed significant anti-cancer activities (inhibitory rate>35%) and were devoid of toxic effects. Compound 29 showed the highest inhibitory rate at 69.25%, indicating that it might be a promising lead compound. PMID- 15351401 TI - Benzimidazole derivatives. Part 5: design and synthesis of new benzimidazole arylpiperazine derivatives acting as mixed 5-HT1A/5-HT3 ligands. AB - A series of new mixed benzimidazole-arylpiperazine derivatives were designed by incorporating in general structure III the pharmacophoric elements of 5-HT(1A) and 5-HT(3) receptors. Compounds 1-11 were synthesized and evaluated for binding affinity at both serotoninergic receptors, all of them exhibiting high 5-HT(3)R affinity (K(i)=10-62nM), and derivatives with an o-alkoxy group in the arylpiperazine ring showing nanomolar affinity for the 5-HT(1A)R (K(i)=18-150nM). Additionally, all the synthesized compounds were selective over alpha(1) adrenergic and dopamine D(2) receptors (K(i)>1000-10,000nM). Compound 3 was selected for further pharmacological characterization due to its interesting binding profile as mixed 5-HT(1A)/5-HT(3) ligand with high affinity for both receptors (5-HT(1A): K(i)=18.0nM, 5-HT(3): K(i)=27.2nM). In vitro and in vivo findings suggest that this compound acts as a partial agonist at 5-HT(1A)Rs and as a 5-HT(3)R antagonist. This novel mixed 5-HT(1A)/5-HT(3) ligand was also effective in preventing the cognitive deficits induced by muscarinic receptor blockade in a passive avoidance learning test, suggesting a potential interest in the treatment of cognitive dysfunction. PMID- 15351402 TI - Structure-activity relationship of phosmidosine: importance of the 7,8-dihydro-8 oxoadenosine residue for antitumor activity. AB - To study the structure-activity relationship of phosmidosine, a variety of phosmidosine derivatives 9a-g were synthesized by condensation of N-diisopropyl N'-(N-tritylprolyl)phosphorodiamidite 6 with appropriately protected nucleoside derivatives 7a-g. As the result, replacement of the 7,8-dihydro-8-oxoadenine base by adenine and 6-N-acetyladenine did not affect the antitumor activity. However, phosmidosine derivatives containing uracil, cytosine, and guanine in place of the 7,8-dihydro-8-oxoadenine base did not show significant activity. A plausible explanation for the selective expression of phosmidosine compared with that of phosmidosine analogs having other amino acids in place of proline is also discussed. These results suggest that phosmidosine serves as an inhibitor of prolyl adenosine 5'-phosphate (prolyl-AMP) to inhibit the peptide synthesis in cancer-related cells. PMID- 15351403 TI - Peptidyl allyl sulfones: a new class of inhibitors for clan CA cysteine proteases. AB - A new series of peptidyl allyl sulfone inhibitors was discovered while trying to synthesize epoxy sulfone inhibitors from vinyl sulfones using basic oxidizing conditions. The various dipeptidyl allyl sulfones were evaluated with calpain I, papain, cathepsins B and L, cruzain and rhodesain and found to be potent inhibitors. In comparison to the previously developed class of vinyl sulfone inhibitors, the novel dipeptidyl allyl sulfones were more potent inhibitors than the corresponding dipeptidyl vinyl sulfones. It was observed that the stereochemistry of the vinyl sulfone precursor played a role in the potency of the dipeptidyl allyl sulfone inhibitor. PMID- 15351404 TI - Synthesis and biological properties of novel glucocorticoid androstene C-17 furoate esters. AB - A series of novel corticosteroid derivatives featuring C-17 furoate ester functionality have been synthesised. Profiling in vitro and in vivo has resulted in the identification of a compound with a longer duration of action and a lower oral side effect profile in rodents compared to budesonide. PMID- 15351405 TI - Analysis of human topoisomerase I inhibition and interaction with the cleavage site +1 deoxyguanosine, via in vitro experiments and molecular modeling studies. AB - Human topoisomerase I (Top1) plays a pivotal role in cell replication and transcription, and therefore is an important anti-cancer target. Homocamptothecin is a lead compound for inhibiting Top1, and is composed of five conjugated planar rings (A-E). The homocamptothecin E-ring beta-hydroxylactone opens slowly to a carboxylate at pH>7.0. We analyzed, which form of homocamptothecin was biochemically relevant in the following ways: (1) the homocamptothecin carboxylate was tested for activity in vitro and found to be inactive; (2) homocamptothecin was incubated with Top1 and dsDNA, and we found that the homocamptothecin beta-hydroxylactone form was stabilized; (3) the homocamptothecin E-ring beta-hydroxylactone was modified to prevent opening, and the derivatives were either inactive or had low activity. These results indicated that the homocamptothecin beta-hydroxylactone was the active form, and that an E ring carbonyl oxygen and adjacent unsubstituted/unprotonated ring atom were required for full activity. Homocamptothecin and derivatives were docked into a Top1/DNA active site model, in which the +1 deoxyguanosine was rotated out of the helix, in order to compare the interaction energies between the ligands and the Top1/DNA active site with the in vitro activities of the ligands. It was found that the ligand interaction energies and in vitro activities were correlated, while the orientations of the ligands in the Top1/DNA active site explained the importance of the E-ring beta-hydroxylactone independently of E-ring opening. An essential component of this Top1/DNA active site model is the rotated +1 deoxyguanosine, and in vitro experiments and molecular modeling studies supported rotation of the +1 deoxyguanosine out of the helix. These results allow for the rational design of more potent Top1 inhibitors through engineered interactions with as yet unutilized Top1 active-site residues including: Glu356, Asn430, and Lys751. PMID- 15351406 TI - UKR--time for caution? PMID- 15351407 TI - Minimally invasive unicompartmental knee arthroplasty. PMID- 15351408 TI - Medium-term results of the AMC-unicompartmental knee arthroplasty. AB - This study presents the outcome of patients treated by AMC-unicompartmental knee arthroplasty (UCA). The prosthesis was implanted in 361 cases for the medial or lateral compartment arthritis. The mean follow-up period was 5.5 (range: 2.3 12.5) years. Mean age at surgery was 69.5 (range: 46.3-88.6) years. Six knees were lost to follow-up (1.6%). Patients were assessed by an independent clinical observer and results were standardised using the Knee Society Rating System. The roentgenographic analyses were performed according to the Knee Society Evaluation System. Ninety-five percent of patients had no pain or slight pain at the latest follow-up, 92% had a good or excellent clinical outcome. Three knees were revised for mobile bearing dislocation after medial UCA and three for lateral mobile bearing dislocation after lateral UCA. Five revisions were necessitated by component loosening and there was one case of deep infection. The clinical results of the investigated patients demonstrate that the AMC-UCA is a successful concept with safe fixation of the prosthesis and good durability of the mobile bearings. PMID- 15351409 TI - Comparison of a mobile with a fixed tibial bearing unicompartimental knee prosthesis: a prospective randomized trial using a dedicated outcome score. AB - Between February 1996 and December 1997, forty consecutive patients with medial compartment knee arthritis were assigned randomly into two groups. All underwent a unicompartimental knee replacement. The tibial prosthesis in group A (20 knees) had a fixed bearing and in group B (20 knees) a mobile bearing. The average age was 69. Five in group A and 71 in group B. Pre-operatively, and at an average follow-up of 5.7 years, all the knees were assessed using both a dedicated UKR score specifically developed and adopted by the Italian Orthopaedic Unicompartimental Knee Replacement Users Group (GIUM) and the Knee Society Score. During surgery in group B we experienced a medial tibial plateau fracture fixed intra-operatively. One prosthesis in group A required an early revision to a TKR because of a continuous pain without any evident sign of loosening. There were no meniscal dislocations in group B. No statistically significant difference in outcome was observed between the two groups at the latest follow-up. PMID- 15351410 TI - Minimally invasive Oxford unicompartmental knee arthroplasty: functional results at 1 year and the effect of surgical inexperience. AB - Oxford medial unicompartmental arthroplasty (UCA) is now routinely performed through a short incision without dislocation of the patella. We present the 1 year results of the first 104 knees operated in Oxford using this technique and the learning curve associated with it. The patient's average age at surgery was 68 years. The average American Knee Society 'knee score' improved from 37 to 94 points and the average 'functional score' from 50 to 92 points. Average maximum flexion improved from 117 to 131 degrees. The average knee score during a surgeon's first 10 cases was 88 points, which was significantly less (P<0.03) than the score achieved for subsequent cases after the first 10 (95 points). These data suggest for the first time that a learning curve exists with worse results being achieved during a surgeon's first 10 cases. However, the results during the learning curve are still similar to that achieved after total knee arthroplasty. PMID- 15351411 TI - Unicompartmental knee arthroplasty: an intermediate report of survivorship after the introduction of a new system with analysis of failures. AB - We describe the outcome of a series of 66 consecutive porous coated low contact stress (LCS) unicompartmental knee arthroplasty (UKA) cases performed in 52 patients for osteoarthritis (OA) by a single surgeon. Both survival, using the endpoint of revision for any cause, and knee function, using the Oxford knee score (OKS) as a validated outcome measure, were established in a retrospective review. At an average postoperative follow-up period of 5.9 years (range 5.1 6.6), there were 8 knees in patients who had died and 58 knees in those who were still living. We established the status of all knees, and prosthesis survival at 5 years was 89.7% (95% confidence interval, 81.6% to 97.7%). Technical errors were responsible for four of six failures and included progression of lateral compartment OA due to overcorrection, a medial tibial stress fracture due to poor pin placement, and a case where cement was required and poor cementing technique lead to early tibial component loosening. In the remaining 52 knees, the average preoperative OKS had improved significantly (p<0.0001) from 37.0 (range, 17-49) to a postoperative score of 20.5 (range, 13-32). We conclude that the functional results following UKA compare favourably to total knee arthropasty (TKA); however, the survivorship of this series does not match that of published reports of TKA. The introduction of a new system of UKA includes the risk of early failures due to surgeon error, even when a surgeon is competent in UKA, warranting careful surveillance during this period. PMID- 15351412 TI - Analysis of first forty Oxford medial unicompartmental knee replacement from a small district hospital in UK. AB - We present a prospective review of our first 40 consecutive Oxford unicompartmental knee replacements for anteromedial osteoarthritis from a small District General Hospital performing an average of 8 operations each year. At an average follow-up of 7.5 years (6-10 years), no revisions were undertaken with a significant improvement in the Oxford and American Knee Score at review. Our study confirms the good results reported from other centres when strict selection criteria and surgical techniques are met. It does not support the observation that less number of these procedures performed each year correlate with a poorer outcome. PMID- 15351413 TI - Fixed or mobile bearing unicompartmental knee replacement? A comparative cohort study. AB - Unicompartmental knee replacement (UKR) for anteromedial osteoarthritis is a well established technique. Numerous clinical outcome studies demonstrate good medium and long-term results. The aim of this prospective study was to compare the complications and short-term clinical outcome of the St. Georg Sled, a fixed bearing UKR, with the Oxford mobile bearing UKR. Outcome at 2 years was assessed using the Bristol knee score and the Oxford knee score; in addition, complications occurring since formal scoring at 2-year follow-up were recorded. A total of 47 Oxford UKRs and 57 St. Georg Sled UKRs were performed in 91 patients, none of whom were lost to follow-up. At 2 years, both outcome measures were better for the St. Georg Sled Group. The pain component of the Bristol knee score was significantly better for this group (p-value = 0.013). Three patients in the Oxford group had bearing dislocations and a further four patients required revision (mean time to revision 3.0 years). In the St. Georg Sled group, three patients required revisions (mean time to revision 3.4 years). These results demonstrate that in the short-term, the Oxford mobile bearing prosthesis has a higher re-operation rate and that the St. Georg Sled achieves better pain relief. The functional scores of the two groups were similar. PMID- 15351414 TI - The effect of tibial implant size on post-operative alignment following medial unicompartmental knee replacement. AB - We have retrospectively reviewed a consecutive series of 71 medial unicompartmental knee replacements in 56 patients performed between 1991 and 1996 by a single surgeon. The aim of this study was to quantify the amount of correction in the tibiofemoral angle, which is achievable with each thickness of polyethylene tibial insert. Our results have shown that the amount of correction which can be expected with an 8 mm, 10 mm, 12 mm and 14 mm insert are 5.3 degrees, 4.8 degrees, 6.6 degrees and 9.5 degrees, respectively. These results will act as a useful reference for surgeons who perform this procedure. PMID- 15351415 TI - Early complications of unicompartmental knee replacement: the Droitwich experience. AB - A total of 185 patients underwent unicompartmental knee replacements (UKR) in 206 knees over a period of 5 years. This is a retrospective look at the early results and a comment on the complications seen within the first year in these patients. Patient selection and technical error were responsible for the bulk of the complications seen within the first year post-operatively. PMID- 15351416 TI - The precision and accuracy of templating the size of unicondylar knee arthroplasty. AB - In trauma and joint arthroplasty, preoperative templating is an important step that can help in the selection of implant size, position and alignment. Although the precision (reproducibility) of templating in unicondylar knee arthroplasty has been assessed previously, the accuracy has never been studied. Our aim is to assess the precision and accuracy of using the templating system for one commonly used unicondylar knee arthroplasty, ALPHANORM design (Alphanorm Medizintechnik, Germany). Eight observers used the templating system to estimate the size of the unicondylar knee prosthesis ALPHANORM in 29 randomly selected patients with osteoarthritis. The observers, who all were orthopaedic surgeons with two different levels of experience, worked independently and repeated their measurements 2 weeks later. All the patients subsequently underwent unicondylar knee replacements, and the actual size of the femur and tibia was assessed intra operatively without any knowledge of the template sizes. Our results revealed a high level of intra-observer reproducibility. However, the inter-observer reproducibility and the accuracy all were poor. PMID- 15351417 TI - Is closed-suction drain necessary in unicompartmental knee replacement? A prospective randomised study. AB - In a prospective randomised trial we evaluated the use of a post-operative closed suction drain in unicompartmental knee replacement (UKR). Seventy-eight patients were divided into two groups: one without a post-operative closed-suction drain (Group A) and one with a drain (Group B). Both groups were matched for age, sex and pre-operative haemoglobin. In group A we observed a lower day one post operative analgesic requirement, smaller knee circumference 3 days post operatively and less local wound complications. Drain usage in UKR resulted in no significant advantage in post-operative pain, range of motion and hospital stay. Post-operative drainage does, however, increase the cost of the procedure both in labour and equipment expenditure. No deep infections occurred in either group during the follow-up period. We conclude that avoiding post-operative closed suction drainage in UKR does not influence the outcome. PMID- 15351418 TI - Radiological changes ten years after St. Georg Sled unicompartmental knee replacement. AB - Failure of a unicompartmental knee replacement (UKR) may be caused by progressive osteoarthritis of the knee and/or failure of the prosthesis. We have examined the fate of the other compartments of the knee by radiological assessment 10 years after operation. A total of 50 UKRs were performed on 45 carefully selected patients between 1989 and 1992. Fifteen patients died, two patients were lost to follow-up and two knees were revised. Standard long-leg weight-bearing anteroposterior views of the knee and skyline views of the patellofemoral joint were taken pre-operatively and at 8 months and 10 years after operation. The radiographs of the remaining 30 knees were reviewed three times by blind and randomized assessment to measure the progression of osteoarthritis within the joints. Two knees showed evidence of progression of osteoarthritis within the patellofemoral joint and three knees showed some progression of the opposite tibiofemoral compartment--but only on one of the radiological grading systems used for assessment. It is concluded that progression of arthritis in the unreplaced compartments is not a significant problem after fixed bearing UKR. PMID- 15351419 TI - Wear debris associated with a large lateral femoral condyle cyst following an Oxford medial unicompartmental knee replacement. AB - A 63-year-old gentleman presented with a history of worsening left knee pain almost 5 years following bilateral uni-compartmental knee replacements. X-Rays revealed a fracture of the medial tibial plateau and revision surgery was undertaken. A large cyst was found in the lateral femoral condyle, which extended from the weight bearing area in the lateral compartment to the margin of the trochlea. Histology of the lesion revealed a central degenerate cyst with multiple particles of polyethylene and cement debris around its periphery. Cyst progression in association with wear debris is a rare problem following unicompartmental knee replacement. The findings at the time of revision in this case demonstrate that the complications of unicompartmental arthroplasty may affect all compartments of the knee. PMID- 15351420 TI - Spontaneous recurrent haemarthrosis following unicompartmental knee arthroplasty and its successful treatment by coil embolisation. AB - A case of spontaneous recurrent haemarthrosis of the knee presenting 3 months after the unicondylar knee replacement is described. Femoral arteriography demonstrated hypertrophy of the saphenous branch of descending genicular artery with a prominent vascular blush in the region of posteromedial aspect of the popliteal fossa thought to represent a hypertrophic vascular mass of synovium. Therapeutic embolisation of the saphenous branch of descending genicular artery was performed using three fibred platinum coils with satisfactory clinical results. Since the embolisation, the haemarthrosis has not recurred. To our knowledge, this therapeutic procedure used to treat recurrent haemarthrosis following unicondylar knee arthroplasty has not been previously described in the literature. PMID- 15351421 TI - The congenital muscular dystrophies in 2004: a century of exciting progress. AB - The congenital muscular dystrophies are a heterogeneous group of inherited disorders. The clinical features range from severe and often early fatal disorders to relatively mild conditions compatible with survival into adult life. The recent advances in the genetic basis of congenital muscular dystrophies have allowed to significantly improve our understanding of their pathogenesis and clinical diversity. These advances have also allowed to classify these forms according to a combination of clinical features and primary biochemical defects. In this review we present how the congenital muscular dystrophies field has evolved over the last decade from a clinical and genetic point of view. PMID- 15351422 TI - The role of muscle biopsy in analysis of the dystrophin gene in Duchenne muscular dystrophy: experience of a national referral centre. AB - Although the majority (65%) of boys with Duchenne muscular dystrophy (DMD) carry a deletion in the dystrophin gene, finding mutations in the remaining families is vital for counselling. We have provided a comprehensive mutation service as a national referral centre for France for over 10 years and we report here our experience. Mutation screening is on mRNA from a muscle biopsy. We have detected 79 mutations in 89 samples referred with a diagnosis of DMD, which is the most comprehensive survey to date of the full range of nondeletion mutations. Although some mutations were nonsense mutations, some frameshift mutations and some splicing mutations, all of them led to the generation of premature stop codons or a shortened product which could be detected using the Protein Truncation Test. We recommend a protocol which is robust and sensitive applied to the entire coding region reverse-transcribed from dystrophin transcripts from muscle biopsy. PMID- 15351423 TI - Mutations in Czech LGMD2A patients revealed by analysis of calpain3 mRNA and their phenotypic outcome. AB - Calpain3 (CAPN3, p94) is a muscle-specific nonlysosomal cysteine proteinase. Loss of proteolytic function or change of other properties of this enzyme (such as stability or ability to interact with other muscular proteins) is manifested as limb girdle muscular dystrophy type 2A (LGMD2A, calpainopathy). These pathological changes in properties of calpain3 are caused by mutations in the calpain3 gene. The fact that the human gene for calpain3 is quite long led us to analyse its coding sequence by reverse transcription-PCR followed by sequence analysis. This study reports nine mutations that we found by analysing mRNA of seven unrelated LGMD patients in the Czech Republic. Three of these mutations were novel, not described on the Leiden muscular dystrophy pages so far. Further, we observed a reduction of dysferlin in muscle membrane in five of our seven LGMD2A patients by immunohistochemical analysis of muscle sections. PMID- 15351424 TI - Insulin sensitivity and skeletal muscle glucose transport in horses with equine polysaccharide storage myopathy. AB - Equine polysaccharide storage myopathy (PSSM) is an inherited disorder characterized by the accumulation of glycogen and abnormal polysaccharide in muscle with normal glyco(geno)lytic enzyme activities. The purpose of this study was to evaluate in vivo insulin sensitivity and glucose excursion in PSSM using a euglycemic hyperinsulinemic clamp. In addition, the content of muscle glucose transporters (GLUT1 and GLUT4) and the insulin receptor was determined in muscle biopsies using Western blot analysis. The glycogen content was 1.8-fold higher, and isolated polysaccharide analyzed by iodine absorption spectra, was less branched in equine PSSM. Throughout the clamp, the affected horses required a higher rate of glucose infusion to maintain euglycemia. Although GLUT1 content was lower, the total content of GLUT4 and insulin receptor was not different in myopathic vs. control horses. PSSM therefore represents a novel disorder where enhanced insulin sensitivity and elevated glucose excursion leads to increased synthesis of muscle glycogen, which in our horses appears to be independent of augmented GLUT4 or IR quantity. PMID- 15351425 TI - Histological parameters for the quantitative assessment of muscular dystrophy in the mdx-mouse. AB - Duchenne muscular dystrophy is a severe X-linked hereditary disease caused by the absence of functional dystrophin. The dystrophin-deficient mdx-mouse strain is a widely used animal model for dystrophin-deficiency. Several therapeutic approaches for muscular dystrophy have been proposed by different laboratories. In order to compare the efficacy of these therapies in the mdx-mouse, it is essential to implement standardized protocols for the assessment of functional and histological parameters in this mouse model. Here, we determine that the minimal 'Feret's diameter' is a geometrical parameter that allows for reliable measure of muscle fiber cross-sectional size. Using this geometrical parameter we calculate variance coefficients of the muscle fiber size and provide reference values for the quantitative assessment of dystrophic symptoms in frequently investigated muscles of wild-type and mdx-mouse. In addition, we compare the variance coefficients of the muscle fiber size with the percentage of muscle fibers with centralized nuclei; another histological hallmark of muscular dystrophy. PMID- 15351426 TI - Increased risk for cardiorespiratory failure associated with the A3302G mutation in the mitochondrial DNA encoded tRNALeu(UUR) gene. AB - Screening the mitochondrial DNA of a 64-year-old woman with mitochondrial myopathy revealed 76% of the tRNA(Leu(UUR)) A3302G mutation in muscle. Muscle of her affected son carried 96% mutated mitochondrial DNA. Both patients were biopsied twice, showing isolated complex I deficiency in the son's first biopsy, additional increased (within normal range) complex II + III activities in his second biopsy, combined complex I, II + III deficiency in mothers first biopsy and additional complex IV deficiency in her second biopsy. After a stay in the mountains, the son died of cardiac arrhythmia. The A3302G mutation has been reported before and is associated with mitochondrial myopathy and cardiorespiratory failure. Pathogenesis is explained by abnormal mtRNA processing, which was also reported for the adjacent C3303T mutation associated with cardiomyopathy and/or skeletal myopathy. Our findings suggest that a high mutation load of the A3302G mutation can lead to fatal cardiorespiratory failure, likely triggered by low environmental oxygen pressure and exercise. PMID- 15351427 TI - Expanding the phenotype of potassium channelopathy: severe neuromyotonia and skeletal deformities without prominent Episodic Ataxia. AB - We report an unusual family in which the same point mutation in the voltage-gated potassium channel gene KCNA1 resulted in markedly different clinical phenotypes. The propositus presented in infancy with marked muscle stiffness, motor developmental delay, short stature, skeletal deformities, muscle hypertrophy and muscle rippling on percussion. He did not experience episodic ataxia. His mother presented some years later with typical features of Episodic Ataxia type 1 (EA1), with episodes of ataxia lasting a few minutes provoked by exercise. On examination she had myokymia, joint contractures and mild skeletal deformities. A heterozygous point mutation in the voltage-gated K(+) channel (KCNA1) gene (ACG AGG, Thr226Arg) was found in both. We conclude that mutations in the potassium channel gene (KCNA1) can cause severe neuromyotonia resulting in marked skeletal deformities even if episodic ataxia is not prominent. PMID- 15351428 TI - Subclinical cardiac involvement in myotonic dystrophy. PMID- 15351430 TI - Prednisone in dystrophin-deficient Caernorabditis elegans. PMID- 15351432 TI - A glycogen synthase kinase 3-beta promoter gene single nucleotide polymorphism is associated with age at onset and response to total sleep deprivation in bipolar depression. AB - The molecular mechanisms driving the biological clock in the suprachiasmatic nucleus of the hypothalamus may play a role in mood disorders. A single nucleotide polymorphism (SNP) (-50T/C) falling into the effective promoter region (nt -171 to +29) of the gene coding for glycogen synthase kinase 3-beta (GSK3 beta) has been linked with different age at onset of bipolar illness. GSK3-beta codes for an enzyme which is a target for the action of lithium and valproic acid, and the inhibition of which causes antidepressant-like behaviors in a preclinical model. We studied the effect of this polymorphism on the acute response to total sleep deprivation of 60 depressed bipolar type I inpatients. Homozygotes for the mutant allele of GSK3-beta promoter (-50T/C) SNP showed a later onset of bipolar illness, and better acute effects of TSD treatment on perceived mood (as rated on VAS). Overall, these observations suggest a protective role for this genotype in respect to bipolar illness. Results warrant interest for the variants of genes pertaining to the molecular clock as possible endophenotypes of bipolar disorder, and for GSK3-beta as a target of a new class of antidepressant drugs, but caution ought to be taken in interpreting these preliminary results and future replication studies must be awaited because of the low frequency of the GSK3-beta*C/C genotype in the studied populations. PMID- 15351433 TI - Distribution of pH regulators in the rat laryngeal nerve: the spatial relationship between Na(+)/HCO(3)(-) cotransporters and Na(+)/H(+) exchanger type 3. AB - We studied the distribution of Na(+)/HCO(3)(-) cotransporters (NBC) and Na(+)/H(+) exchanger type 3 (NHE3) in the laryngeal nerve by immunohistochemistry to elucidate the spatial relationship of pH regulation system in the peripheral nerves. The nervous components, i.e., the nerve cells in the nodose and local ganglia and nerve fibers, were immunoreactive for NBC. Glial components such as Schwann cells and satellite cells surrounding nerve fibers and nerve cell bodies were also immunoreactive for NBC in most cases, while the cells comprising the perineurium and endoneurium were immunoreactive for NHE3. These results suggest that NBC-dependent pH regulation systems are present in the laryngeal nerve. Whereas, NHE3 may regulate extracellular pH rather than intracellular pH. In conclusion, spatial relationship of NBC and NHE3 in the laryngeal nerve would be important for pH regulation. Perineural lymph may have key role for acid-induced modulation of axons and Schwann cells. PMID- 15351434 TI - Phase-resetting response to (+)8-OH-DPAT, a serotonin 1A/7 receptor agonist, in the mouse in vivo. AB - Several lines of evidence indicate that 5-HT neuronal systems may play a critical role for the non-photic entrainment of the rodent circadian clock. Although it is well established that (+)8-hydroxy-2-(di-n-propylamino)tetralin [(+)8-OH-DPAT], a 5-HT(1A/7) receptor agonist, causes a phase-advance of behavioral rhythm in hamsters, little is known whether this agent produces phase shifts of activity rhythm in mice. Therefore, we examined the effect of (+)8-OH-DPAT on the mouse locomotor activity rhythm. Systemic administration of this chemical at mid subjective daytime induced a clear and dose-dependent phase advance, while there were no significant phase shifts at other times (early-subjective day, late subjective day, or subjective night). Additionally, (+)8-OH-DPAT accelerated the re-entrainment of mouse behavioral rhythm to a 6-h advanced light-dark cycle. These results suggest that we can use mice for understanding the molecular mechanism of (+)8-OH-DPAT-induced phase shift because of availability of clock gene targeted mice. PMID- 15351435 TI - Regional changes in glial cell line-derived neurotrophic factor after cardiac arrest and hypothermia in rats. AB - Hypothermia after resuscitation from cardiac arrest reduces functional and histological brain injury. Stimulation of neurotrophic factors may contribute to the beneficial effects of hypothermia. This study examined the effects of cardiac arrest and induced hypothermia on regional levels of glial cell line-derived neurotrophic factor (GDNF) over the first 24 h after rat cardiac arrest. Hypothermia increased GDNF in hippocampus at 6 h, but did not prevent a subsequent decline in hippocampal GDNF. In contrast, hypothermia prevented early increases in cortical levels of GDNF at 3 and 6 h. Cerebellar GDNF increased slightly over 24 h in hypothermia-treated rats, but brainstem levels of GDNF did not change in response to cardiac arrest or hypothermia. These results suggest that temperature after resuscitation produces regionally specific changes of GNDF levels in brain. PMID- 15351436 TI - Influence of polymorphisms in the genes for cytokines and glutathione S transferase omega on sporadic Alzheimer's disease. AB - We studied promoter region polymorphisms in the interleukin (IL)-1alpha, IL 1beta, IL-6, IL-10, tumor necrosis factor, and transforming growth factor (TGF) beta1 genes in Japanese patients with Alzheimer's disease (AD) (n = 172) and normal controls (n = 163). We also examined an association of a polymorphism located in the glutathione S-transferase omega 1 (GSTO-1) gene region with AD patients. None of these genotypes or allele frequencies showed a significant difference between AD patients and controls. We also failed to detect any difference in the disease onset between each genotype of the seven polymorphisms. Although AD patients carrying high producer alleles of TGF-beta1 and IL-1beta or TGF-beta1 and IL-6 showed a tendency for an early onset of the disease, neither of these combined effects reached a significant level after multiple comparisons. Our findings suggest that genetic polymorphisms in the cytokines and GSTO do not play a major role in Japanese AD patients. PMID- 15351437 TI - Apolipoprotein E receptor 2 is involved in neuritic plaque formation in APP sw mice. AB - Apolipoprotein E receptor 2 (apoER2) is a receptor for apolipoprotein E containing lipoprotein and also for Reelin. Apolipoprotein E-associated risk of developing Alzheimer's disease (AD) may be related to its binding to and clearance by cell surface receptors, including members of the low-density lipoprotein receptor family. Otherwise there is circumstantial evidence that the Reelin signaling pathway may contribute to neurodegeneration in AD. To investigate the role of apoER2 on amyloid deposition and neurodegeneration in vivo, we examined the presence of apoER2 in the brains of APP sw transgenic mice (Tg2576) using three apoER2 monoclonal antibodies. Our immunohistochemical study revealed that apoER2 was localized in fine granular structure and reactive astrocytes surrounding amyloid plaques. The double labeling immunohistochemistry revealed that this granular structure overlaps synaptophysin-positive dystrophic neurites. These findings indicate that neuronal apoER2 may play a role for amyloid deposition and neuronal degeneration in AD. PMID- 15351438 TI - APP and PS-1 mutations induce brain oxidative stress independent of dietary cholesterol: implications for Alzheimer's disease. AB - Epidemiological and biochemical studies strongly implicate a role for cholesterol in the pathogenesis of Alzheimer's disease (AD). Mutation in the PS-1 and APP genes, which increases production of the highly amyloidogenic amyloid beta peptide (Abeta42), is the major cause of familial AD. The AD brain is under significant oxidative stress, including protein oxidation and lipid peroxidation. In the present study, protein oxidation and lipid peroxidation were compared in the brain homogenates from knock-in mice expressing mutant human PS-1 and APP in relation to the intake of dietary cholesterol. The APP and PS-1 mice displayed increased oxidative stress as measured by protein oxidation and lipid peroxidation, independent of dietary cholesterol. These results are discussed with reference to proposed therapeutic strategies of AD. PMID- 15351439 TI - Heterogeneous response of cerebral blood flow to hypotension demonstrated by laser speckle imaging flowmetry in rats. AB - The response of cerebral blood flow (CBF) to mild hypotension shows great variability. CBF changes in different cortical regions at a mean arterial blood pressure (MABP) of 70 mmHg (near the lower limit of CBF-pressure autoregulation) were analyzed using laser speckle imaging flowmetry (LSIF). We hypothesize that variations in CBF autoregulation might be due to differences between regions within the cortex of each animal, regional heterogeneity, as opposed to differences between animals. Different responses of local CBF to hypotension in each rat were demonstrated as %CBF(70), the ratio of CBF at a MABP of 70 mmHg to CBF at a MABP of 100 mmHg. Regional heterogeneity ranged from 12 to 51%, expressed as the coefficient of variation of %CBF(70). At the same time LSIF revealed distinctly different patterns of CBF-pressure autoregulation between animals. Between-animal heterogeneity represents a continuous distribution, spread between animals with predominantly decreased CBF (%CBF(70) < 85%, rat #3) and animals with a high proportion of increased flow responses (%CBF(70) > 115%, rat #2). There are approximately equal contributions to the heterogeneity of %CBF(70) from within-animal (57 +/- 5%, percent variance +/- standard error) and between-animal (43 +/- 26%) variations. Within-animal variations could be due to heterogeneity of vascular anatomy, while the diversity of vascular control mechanisms might contribute to between-animal variations. PMID- 15351440 TI - How the human auditory system treats repetition amongst change. AB - Occasional repetitions embedded in a sequence of ever-changing sounds elicit a frontally negative event-related potential peaking between 100 and 200 ms from the onset of sound repetition (repetition negativity, RN). Three interpretations have been suggested for the emergence of RN: (1) RN is a mismatch negativity (MMN) response elicited because the auditory system detected sound change as a regular feature of the stimulus sequence, which is violated by repetitions. (2) RN reflects short-term sensitization of the auditory system to the specific acoustic parameters of each sound, a process subserving the formation of durable memory traces. (3) The difference between responses elicited by repetition and change reflects differential refractoriness of the P2 component. In order to distinguish between the MMN-based and the other two explanations of RN, we varied the global sequential probability of tone-repetitions (at 5, 20, and 50%) in sequences composed of tones of five different frequencies. Whereas all three explanations of the RN predict a local repetition-probability effect on the RN amplitude, MMN should also be affected by the global repetition-probability, because MMN is sensitive to the global deviant-probability, independently of local deviant-probability. To eliminate differences in local repetition probabilities, we compared the responses elicited by tones that followed the same micro-sequence (change-change-repetition) across the three different global probability conditions. Global repetition probability had a significant effect on RN: it was only elicited with 5%, but not with the two higher repetition probabilities. This result supports the MMN interpretation of the RN. PMID- 15351441 TI - Expression of tumor necrosis factor alpha in nerve fibers and oligodendrocytes after transient focal ischemia in mice. AB - The expression of tumor necrosis factor alpha (TNFalpha) increases and participates in several central nervous system (CNS) disorders. However, its expression after transient middle cerebral artery occlusion (tMCAO) in mice is not fully discussed yet. Therefore, we examined gene expression and protein localization of TNFalpha in brain using real-time polymerase chain reaction (PCR) and immunostaining after 1 h tMCAO in mice. After 1 h of ischemic conditions, we observed an increase in the expression of TNFalpha mRNA from basal level. While the expression decreased immediately to control level after reperfusion, it increased again significantly at 24 and 48 h after tMCAO. TNFalpha-like immunoreactivity (TNFalpha-LI) was slightly detected in fibrous structures of the neurons before ischemia. After ischemia, TNFalpha-LI spread widely to the soma of neurons and became more abundant in the nerve fibers, including axonal and dendritic processes. Moreover, TNFalpha-LI was also expressed in the oligodendrocytes and, occasionally, in microglia/macrophages, but not in astrocytes 24 h after tMCAO. These results suggest that TNFalpha shows biphasic expression that corresponds with ischemia and reperfusion, and might play a role in various cells to regulate CNS disorders such as neuronal and oligodendritic cell death after transient ischemia. PMID- 15351442 TI - Chronic treatment with lithium increases the ecto-nucleotidase activities in rat hippocampal synatosomes. AB - Lithium is a mood-stabilizing treatment used in bipolar and other psychiatric disorders. The molecular mechanisms underlying lithium action remain poorly understood. Adenosine is a neuromodulator that possesses anticonvulsant and neuroprotective properties and the ecto-nucleotidase pathway is a metabolic source of the extracellular adenosine. Here we investigated the effect of lithium on the ecto-nucleotidase pathway in synaptosomes from hippocampus and cerebral cortex of adult rats. Male Wistar rats received standard rat chow with lithium chloride (2.5 mg/g of chow) and NaCl (17 mg/g of chow) during 4 weeks. The serum lithium levels were 1.18 +/- 0.05 mEq./L. ATP and AMP hydrolysis was significantly increased (20 and 35%, respectively) in hippocampal synaptosomes of rats chronically treated with lithium chloride. No significant differences were observed in the hydrolysis of the three nucleotides by cortical synaptosomes. In conclusion, the modulation of the ecto-nucleotidase pathway may be a new explanation for the potential neuroprotective lithium action in hippocampal lesions. PMID- 15351443 TI - Intraocular implantation of DNA-transfected retinal pigment epithelium cells: a new approach for analyzing molecular functions in the newt retinal regeneration. AB - Adult newts can regenerate their entire retinas, even after surgical removal of the neural retina (retinectomy), through transdifferentiation of the retinal pigment epithelium (RPE) cells. To develop a new experimental system for analyzing molecular functions during retinal regeneration of adult newts, we attempted to deliver a foreign gene into RPE cells of retina-less eye-cups in vitro. Here we used pCS2mt-GFP as a reporter construct, and selected Polyfect as a transfection reagent. DNA-transfection appeared to be restricted to the RPE cells of retina-less eye-cups and its efficiency was 0.1-0.2%. We tried to implant RPE-choroid tissue containing DNA-transfected RPE cells into the eye of a host animal. The tissue was placed into the posterior eye-chamber immediately after retinectomy so that the implanted RPE tissue was facing the cornea (i.e., normal orientation). The implant and host RPE regenerated one continuous hybrid neural retina. Ocular sections after 60 days of implantation showed that a small number of cells in the regenerating retina were intensely stained with an anti GFP antibody. Some of those cells were believed to be retinal cells such as ganglion cells, amacrine cells and photoreceptors. The GFP-positive cells in the hybrid regenerating retina could represent clones derived from a single RPE cell. These results indicate that this experimental system could become useful in the study of adult newt retinal regeneration. PMID- 15351444 TI - The relation between force magnitude, force steadiness, and muscle co-contraction in the thumb during precision grip. AB - The aim of this study was to investigate the problem of agonist-antagonist co contracti during a precision force task performed at different force levels. Using a precision grip, seven young adults performed a constant force matching task (10, 22.5, 35, 47.5, and 60% maximum) as accurately as possible (10 trials per force level). Muscle co-contraction in the thumb was monitored by the surface EMG activity of the extensor pollicis longus (EPL) and the flexor pollicis brevis (FPB), and the ratio between those EMG activities (EPL/FPB). Results showed that both EMG activities increased as grip force increased, but the EPL/FPB ratio decreased over the range of force investigated. Force steadiness (as expressed by the coefficient of variation, CV) appeared as a U-shape function of the force level (with maximal steadiness at 22.5%). Separate analyses at each force level showed no correlation between CV and EMG indices. In addition, the contrast between trials with high and low CV revealed no significant difference in terms of our EMG indices. We conclude that muscle co-contraction and grip force steadiness depend on grip force magnitude, but grip force steadiness does not depend on muscle co-contraction. PMID- 15351445 TI - Both N-methyl-D-aspartate (NMDA) and non-NMDA receptors mediate glutamate-induced cleavage of the cyclin-dependent kinase 5 (cdk5) activator p35 in cultured rat hippocampal neurons. AB - Cyclin-dependent kinase 5 (cdk5) regulates crucial neurobiological events, and deregulation of cdk5 has been implicated in several neurodegenerative disorders. The deregulation is suggested to occur due to cleavage of the cdk5 activator protein p35 to a smaller p25 fragment by the calcium-activated protease calpain. Here we have elucidated the role of different calcium-permeable ionotropic glutamate receptors in the induction of p35 cleavage in cultured rat hippocampal neurons. The glutamate receptor agonists glutamic acid, N-methyl-D-aspartate (NMDA), kainic acid, and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) were all able to induce p35 cleavage, in a manner depending on extracellular calcium. The effect of glutamate was mediated by NMDA receptors, as it was prevented by the NMDA antagonist MK-801. Cyclothiazide (CTZ), an inhibitor of AMPA receptor desensitization, enhanced glutamate-induced p35 cleavage. In immature 6-day-old cultures the non-NMDA agonist kainic acid provoked p35 cleavage, whereas glutamate and NMDA were ineffective. The data suggest that both NMDA and non-NMDA receptors are able to induce p35 cleavage. Different factors, such as maturation state of neurons or desensitization properties of non-NMDA receptors, may determine which receptor predominantly mediates the effect of glutamate on p35 cleavage. PMID- 15351446 TI - Constitutive activity of N-beta-alanyl-catecholamine ligase in insect brain. AB - N-Beta-Alanyldopamine (NBAD) is the primary catechol tanning agent precursor in typical brown or yellow insect cuticle. The insect integument enzyme responsible for the synthesis of NBAD was reported to be expressed solely in the epidermis, and only at the time of cuticle sclerotization. However, in this study we demonstrate directly that the enzyme also is expressed in a constitutive manner in the neural system of insects. The requirements and kinetic parameters of the brain-associated enzyme appear similar to those of the epidermis-associated enzyme in Ceratitis capitata. The brain-associated enzyme also was able to catalyze the in vitro synthesis of N-beta-alanylnorepinephrine (NBANE) and beta alanyl derivatives of other biogenic amines. A melanic mutant of C. capitata, niger, was unable to conjugate beta-alanine with dopamine or other amines in either the epidermis or the brain. This result strongly supports the idea that these enzymes actually are expressed from a single gene and that differences in regulation must exist that account for the constitutive expression in the neural system. Similar results were obtained in Drosophila melanogaster and other insects. From these data, a number of questions arise about the role of beta alanyl derivatives of biogenic amines and other compounds in insect brain and similarly, in the mammalian CNS. PMID- 15351447 TI - Effects of food consistency on the pattern of extrinsic tongue muscle activities during mastication in freely moving rabbits. AB - The effects of physical characteristics of foods on the coordination of extrinsic tongue muscle activities during natural mastication were evaluated. Electromyograms of tongue-retractor (styloglossus, SG) and tongue-protractor (genioglossus, GG) muscles as well as the jaw-movement trajectories were recorded during raw rice and chow pellet chewing in the freely moving rabbit. Each masticatory cycle included a jaw closing (Cl) phase consisting of a fast-closing (FC) and a slow-closing (SC) phase, and a jaw opening (Op) phase. The duration of the Cl and SC phases was found to be much larger while the duration of the FC phase was much smaller during rice chewing than pellet chewing. The jaw movements during rice chewing had smaller amplitudes of the gape and lateral excursion of the jaw as compared with those during pellet chewing. The SG muscle had a double peaked burst activity in each masticatory cycle with one peak during the Op phase (the SG1 burst) and the other during the Cl phase (the SG2 burst). They were significantly larger during pellet chewing as compared with rice chewing, but the duration of the SG2 burst was significantly longer during rice chewing than pellet chewing. The offset of the SG2 burst was delayed during rice chewing as compared with that during pellet chewing. There was little difference in the activity pattern of the GG burst between the foods. Our present results suggest that the SG muscle activity could be modified by the sensory feedback possibly to adapt to environmental demands during chewing. PMID- 15351448 TI - Changes to cold detection and pain thresholds following low and high frequency transcranial magnetic stimulation of the motor cortex. AB - There is some evidence that repetitive transcranial magnetic stimulation (rTMS) can alleviate the experience of chronic pain. The mechanisms by which rTMS may induce pain relief, however, are unknown. The present study examined whether a session of rTMS would produce sensory threshold changes in healthy individuals. Detection and pain thresholds for cold sensations were compared following low frequency (1 Hz) (Experiment 1) and high frequency (20 Hz) (Experiment 2) repetitive TMS. While cold detection threshold was significantly lowered by both rTMS rates, only high frequency rTMS produced a significant change in cold pain threshold. In contrast, sham rTMS did not alter thresholds for cold stimuli. These findings provide evidence that sensory thresholds can be influenced by repetitive transcranial magnetic stimulation. PMID- 15351449 TI - Does brain specific 24S-hydroxycholesterol in plasma indicate the disruption of the blood-brain barrier in patients with ischemic stroke? AB - Determination of plasma 24S-hydroxycholesterol, which is produced almost exclusively in the brain and is released only in small amounts into circulation under physiological conditions, might be a marker to monitor non-invasively the time course and the extent of the disintegration of the blood-brain barrier after cerebral ischemia. We investigated the plasma concentrations for 4 to 10 days of 24S-hydroxycholesterol, and compared the concentration with the liver-specific oxysterol 7alpha-hydroxycholesterol, the ubiquitously produced 27 hydroxycholesterol, and cholesterol itself in six patients who were admitted to the hospital within 24 h after symptoms of stroke. Quantification of oxysterols was performed by isotope dilution mass spectrometry and cholesterol by gas-liquid chromatography. Initial concentrations of cholesterol, 24S-, 7alpha-, and 27 hydroxycholesterol in stroke patients were not different from data of healthy controls given in the literature. During the following days, no changes could be observed in the concentrations of cholesterol nor in the other oxysterols. Particularly the brain specific 24S-hydroxycholesterol was very constant and showed only minimal changes. Furthermore, comparison of patients with extended or small lesions did not reveal any differences in the concentrations of oxysterols. Therefore, circulating levels of 24S-hydroxycholesterol are supposed to be only of limited value for monitoring the brain-blood barrier function in patients with acute ischemic stroke. PMID- 15351450 TI - Apparent scarcity of glial fibrillary acidic protein expression in the brain of the pygmy shrew Sorex minutus as revealed by immunocytochemistry. AB - We examined astroglial cells in the brain of the pygmy shrew Sorex minutus (Insectivora). For that purpose we labeled glial fibrillary acidic protein (GFAP) immunohistochemically in brain sections with a polyclonal antibody. Antigen retrieval experiments were performed to counteract formaldehyde fixation masking of GFAP epitopes. Our results showed remarkable paucity of GFAP-immunoreactive cells and fibers in the cerebral cortex and nuclei, as well as in the majority of the diencephalic and mesencephalic structures. In the forebrain, significant numbers of GFAP-containing astrocytes were found only in the ependyma and subventricular zones, superficial part of layer I of the cerebral cortex, and the majority of white matter structures. In the diencephalon, habenular nuclei were rich in GFAP-immunopositive astrocytes and labeled radial fibers were extended between median eminence and the third ventricle. A considerably higher density of labeled astrocytes was detected in the caudal brainstem and cerebellum. In contrast, in the mouse brain, immunoreactive astrocytes were present in large quantities in various structures. Staining of sections of the shrew brain against glutamine synthetase revealed abundance of immunofluorescent astrocytes in many areas, especially in the shrew cerebral cortex. It seems probable that in the shrew brain only a limited fraction of astroglia expresses GFAP, while other astroglial cells can be detected with different markers. It is possible that the rodent type of astroglial GFAP expression might not be common to insectivores and probably to some other mammalian orders. PMID- 15351451 TI - Posttraining administration of pentylenetetrazol dissociates gabapentin effects on memory consolidation from that on memory retrieval process in mice. AB - Gabapentin (GBP), an anticonvulsant drug, 10 mg/kg, i.p., but not 100 mg/kg, i.p., enhanced retention of an inhibitory avoidance task when given 20 min after training, as indicated by retention performance 48 h later. The immediate post training administration of pentylenetetrazol (PTZ, 45 mg/kg, i.p.) impaired retention performance. The amnesic effects of the convulsant drug PTZ were not influenced by GBP at any level of doses. However, GBP 100 mg/kg, but not 10 mg/kg, delayed the latency to first clonic body seizures and decreased the duration of convulsion induced by PTZ. The enhancing effect of GBP on retention was not prevented by the opiate receptor antagonist, naltrexone (0.01 mg/kg, i.p.), which completely prevented the impairment of retention caused by PTZ. Further, naltrexone did not modify the convulsions induced by PTZ. In mice pretreated with naltrexone and that received PTZ, the administration of GBP again, enhanced retention performance during the retention test. Since previous results indicate that the amnesic action of PTZ are due to an effect on memory retrieval, the present results provide additional pharmacological evidence suggesting that GBP influenced memory consolidation and not memory retrieval of an inhibitory avoidance task in mice. PMID- 15351452 TI - Delta and theta power spectra of night sleep EEG are higher in breast-feeding mothers than in non-pregnant women. AB - The power spectra of night sleep EEGs of 12 breast-feeding 9-13 week postpartum mothers were analyzed and compared with those of 12 non-pregnant women. The power spectra in the delta and theta frequency range during NREM sleep for breast feeding mothers were significantly higher than those for non-pregnant women. In addition, the all-night sleep patterns of the mothers were classified into two groups - interrupted sleep due to taking care of their infants and non interrupted sleep - in order to observe the influence of partial sleep deprivation. The power spectra in the delta and theta frequency range were not significantly different between them. This result suggests that increased delta and theta power spectra during postpartum sleep do not result from partial sleep deprivation. The role of prolactin in breast-feeding mothers' sleep is also discussed. PMID- 15351453 TI - Alternative modalities of adenovirus-mediated gene expression in hippocampal neurons cultured on microisland substrate. AB - Previously, we have used CsCl gradient-purified recombinant adenovirus (AdV) to successfully transfer genes into hippocampal neurons cultured on microisland substrate. Here, we report that purification of AdV particles is not required and efficient gene expression can be achieved using either crude AdV lysates or HEK 293 cells infected with AdV. The advantages of the simplified procedure are greatly reduced preparation time and reduced requirements for equipment and expertise. PMID- 15351454 TI - Capsaicin-evoked substance P release in rat dorsal horn increases after peripheral inflammation: a microdialysis study. AB - Numerous in vitro studies suggest that inflammation is associated with enhanced release of substance P (SP) in the dorsal horn. To test the hypothesis that inflammation increases the evoked concentration of SP in the intact animal, we used in vivo microdialysis with a highly sensitive radioimmunoassay to monitor SP like immunoreactivity (SP-LI) in the dorsal horn. Seven days after the induction of persistent unilateral inflammation with hindpaw injection of complete Freund's adjuvant, perfusion of the microdialysis probe with 10 microM capsaicin (a concentration which failed to induce SP-LI release in rats without inflammation) induced a significant increase of microdialysate SP-LI. Inclusion of an NMDA antagonist in the perfusion fluid completely blocked this capsaicin-evoked SP release. Administration of a five-fold higher dose of capsaicin did not further increase SP release. These results in a rat model of chronic arthritis suggest that persistent inflammatory signaling facilitates capsaicin-evoked SP release in the dorsal horn in vivo. PMID- 15351455 TI - The human brain processes visual changes that are not cued by attended auditory stimulation. AB - Event-related potentials (ERPs) to visual stimuli were recorded from the scalp of eight adult humans performing a task in which they counted vowels from a heard story. In the oddball condition, a repeated (standard) light bar of 50 ms in duration was rarely (P = 0.1) replaced by a (deviant) one differing in orientation from the standard. In the control condition, standards were simply omitted from the series and only (alone-) deviants retained. In both conditions, visual stimuli were asynchronous with auditory-task-relevant stimuli. ERPs to deviants significantly differed in amplitude from those to standards in the midline electrodes centrally, parietally and occipitally at 160-200 ms from stimulus onset. Occipitally, such a difference was absent between ERPs to alone deviants and those to standards. The occipital differential ERPs to deviants, which thus could be found only when standards were present in the series, are discussed in the context of the mismatch negativity (MMN). PMID- 15351456 TI - Human neuronal encoding of English syntactic violations as revealed by both L1 and L2 speakers. AB - Our previous study [M. Kubota, P. Ferrari, T.P.L. Roberts, Magnetoencephalography detection of early syntactic processes in humans: comparison between L1 speakers and L2 learners, Neurosci. Lett. 353 (2003) 107-110] showed that an early syntactic response was elicited in first language (L1) speakers for within phrase, but not across-phrase violations, implying that there may exist a continuum of neuronal error gravity. Such an early component was not elicited by second-language (L2) learners. The current auditory study investigated whether two types of different syntactic violations regarding noun-phrase raising (NP raising) and case-filter constructions would elicit a prominent early syntactic component in each hemisphere for both L1 and advanced L2 speakers of English. Neuromagnetic fields were recorded, using a dual 37-channel gradiometer system. A prominent component, peaking at approximately 150 ms post-onset, was observed in both hemispheres of two groups in response to NP-raising induced violations, but not case-filter violations. The findings imply that L1 and L2 speakers have similar neuronal mechanisms subserving syntactic processing of such violations. PMID- 15351457 TI - [Activity report about hospital Saint-Louis's team for taking in charge sequellae of Noma]. AB - A presentation of the country in which the missions have been made, the authors describe the schedule of their action and the synergy developed with associative partners allowing tacking in charge of sequelae of Noma disease. Epidemiology and clinics of Noma are shortly described. Different cases are exposed by localisations: lips, centro-facial, cheek, hemi-face. Different principles of these reconstructions are exposed and the techniques with their adaptation to the African lip and to the particularity of Noma lesion (important lose of substance of mucosae which is very important to repair). The very important points of the organisation are exposed especially for the follow up of these patients. PMID- 15351458 TI - [The retro-orbicularis oculus fat (ROOF) or Charpy's fat pad. Descriptive and functional anatomy. Surgical concepts applied to the design of a frontotemporal lift procedure]. AB - The eyebrow fat pad, or retro-orbicularis oculus fat (ROOF) is a syssarcosis, described as an anatomic and functional unity by M. Charpy in 1909. Our anatomic study on 12 (24 half-head) fresh cadaver specimens confirm anatomic variations, mainly according to age and sex. Synthesis of recent literature shows the renewed interest for this fat pad and its involvement in the eye's lateral flare. We will describe its detailed ultrastructure and the physiopathological mechanisms of aging, causing an earlier lateral eyebrow ptosis. So we can evolve new anatomic concepts applied to recent facial rejuvenative procedures, particularly frontotemporal facelifts (endoscopic or classical procedures), by a harmonious repositioning of the eyebrow and in order to restore the lateral flare of the eye. PMID- 15351459 TI - [Complications of the resurfacing laser: retrospective study of 749 patients]. AB - BACKGROUND: Resurfacing laser is one of the alternative treatment for cutaneous facial rejuvenation. Results are mainly good to excellent, but complications do occur with variable frequency. OBJECTIVES: The purpose of this study was to identify and report the complications they occurred during eight years laser resurfacing and to study their possible avoidance. The author experiences are discussed in conjunction with a review of published reports. METHODS: A retrospective analysis and chart review was performed in 749 patients who underwent cutaneous laser resurfacing with CO2 and Erbium laser by a single operator. Side effects and complications relating to infection, postoperative healing, pigmentary changes, and scarring were tabulated. RESULTS: The most common complication observed was postoperative erythema, which occurred in all patients. Hyperpigmentation was seen in 32% of patients and must be precociously treated. Dermatitis occurred in 10.6% of patients. Hypopigmentation and infections are rare (<1%). CONCLUSION: Although laser resurfacing is a safe and effective method of facial rejuvenation, the cosmetic surgeon must be aware of the various complications that may be encountered. Prompt recognition of complications and appropriate management provide the best opportunity for an acceptable aesthetic outcome. PMID- 15351460 TI - [The lateral fold mammaplasty]. AB - The mammaplasty is characterized by the non-reliability of the shape achieved at the end of the operation: sometimes a conic shape is expected and a relatively flat breast is obtained. The mammaplasty is also characterized by the lateral slipping of the shape with time, cause of the sad aspect of the breast. The purpose of the lateral fold mammaplasty is to facilitate the creation of the desired shape and to stabilize the evolution of this shape with time. The standard mammaplasty technique can determine the sub-mammary fold; the medial contour with its specific shape is already determined. Those inferior and medial folds are stables. But the lateral contour is usually not defined and very rarely stable with time. This procedure creates a well-defined and stable lateral fold, which is the convex junction between the anterior axillary line and the sub mammary fold. This new fold is designed by attachment with five sutures of the axillary sub-dermal tissues to the lateral border of the pectoralis major muscle and just beneath, to the 5th costal arch. A medial tension is applied to the axillary flap before suturing to improve the axillary region. Consequently, the breast shape is more predictable and more stable because the lateral, inferior and medial folds are definitely determined. The realization is simple with no specific complication. It could be an additional procedure to any mammaplasty technique. PMID- 15351461 TI - [Congenital quintus varus supraductus: surgical procedure]. AB - Quintus varus supraductus is a congenital malformation that associates a hyperextension, a varus and an external rotation of the fifth toe coming over the fourth. The goal of this study is to explain an easy, reproducible and efficient surgical procedure to correct that malformation. Our procedure is only about soft tissues. It comprises a fifth toe extensor tenolysis, a circular capsulotomy and lateral ligament section of the fifth metacarpophalangeal articulation. The glenoid plaque is desinserted only if the peroperative reduction did not seem sufficient. The stabilization is achieved through a cutaneous plasty of modified BUTLER, with a cutaneous lengthening VY plasty. Exceptionally, an axial broaching of the articulation has been done. On about 20 children, aged 4-17 years, operated with this procedure, we have noticed a complete and definitive correction in 19 patients. Only one patient showed an incomplete resurgence, but without any functional disturbance. We did not notice any failure. This procedure seems efficient, and all the more interesting that it is technically easy to perform. PMID- 15351462 TI - [Ewing sarcoma of thumb: report of a case and review of the literature]. AB - The authors report a case of an Ewing sarcoma of the thumb. It is a rare localisation of this disease and it should not be unknown. Based on the literature, the appropriate treatment is discussed. PMID- 15351463 TI - [Surgical management of stingray injuries. About two clinical cases]. AB - Fifty injuries by stingrays are annually examined in the New Caledonia hospital. The injuries occur most often in the lower extremity, rare puncture injuries to the thorax or abdomen can cause death. The wound is associated with envenomation. The pain is intense with oedema, cyanosis, erythema often followed by tissue necrosis. In many cases, the management of stingray wounds is inadequate and the gravity often not well appreciated. Localized morbidity and prolonged healing may occur without an effective surgical management. Wound exploration and debridement are particularly indicated after first-aid measures, such as immersion in hot water. The characteristics, treatment and prevention are discussed in connection with two cases. PMID- 15351464 TI - [High frequency ultrasonography and celluscore: an improvement in the objective evaluation of cellulite phenomenon]. AB - As they usually do not have any serious effect on health condition, cellulite phenomenons are not considered as potentially hazardous by general practitioners; unfortunately they involve almost all the feminine population after the forties and may really induce bad side effects either psychological or physical whatever the confusing origin of that cutaneous deformation: heredity, nutrition, circulatory and hormonal diseases etc. Trying to appreciate in a more scientific way this superficial skin disorder, we have developed a computerised questionnaire which can be combined with the finest upto date way of skin exploration, high frequency ultrasonography. It seems possible to consider now cellulite as the result of various disorders which can be separate into fibrotic or retentional phenomenons (surrounding superficial fat tissues) and the direct adipocyte reaction (adiposis). We hope to be able in the future to extend the treatment of cellulite beyond the limits of liposuction which seems until today the only way to bring some improvement to that phenomenon in spite of the very delicacy of the surgical approach. PMID- 15351465 TI - [Management of a patient for mammary reconstruction after mastectomy and lymphadenectomy followed by radiotherapy and chemotherapy]. PMID- 15351466 TI - [Emergency management in a patient wtih an open Gustilo IIIb (Cauchoix III) tibial fracture]. PMID- 15351467 TI - Hospital ethnography: introduction. AB - The introduction sets out two central ideas around which this collection of articles on hospital ethnography has been organised. The first is that hospitals are not identical clones of a global biomedical model. Hospitals take on different forms in different cultures and societies. Medical views and technical facilities may vary considerably leading to different diagnostic and therapeutic traditions. The second idea, related to the first, is that biomedicine and the hospital as its foremost institution is a domain where the core values and beliefs of a culture come into view. Hospitals both reflect and reinforce dominant social and cultural processes of their societies. The authors further discuss some methodological and ethical complexities of doing fieldwork in a hospital setting and present brief summaries of the contributions, which deal with hospitals in Ghana, South Africa, Bangladesh, Mexico, Italy, The Netherlands, Papua New Guinea, Egypt and Lebanon. PMID- 15351468 TI - "Villagers": differential treatment in a Ghanaian hospital. AB - Differential treatment of patients by health workers in African medical institutions is acknowledged by patients, health workers and policy-makers alike to be an obstacle in the realization of government objectives of equity in health care. This qualitative study understands the production and legitimization of differential treatment from the perspective of health workers. On the basis of qualitative field material from a hospital in Northern Ghana the relation between socio-cultural, biomedical and bureaucratic aspects of hospital practice is explored through a focus on categorizations of patients. It is concluded that to blame the "bad attitudes" of health workers for differential treatment is not an adequate explanation. It is important to acknowledge that differential treatment can be understood as a form of agency, and is related to the conditions of hospital work and to the professional and social identities of health workers. PMID- 15351469 TI - The gaps in the gaze in South African hospitals. AB - Analysis of health care systems, especially hospitals, could benefit from Foucault's description of the medical gaze and the panopticon. Foucault's perspective sheds new light on the South African transformation from an oppressive to a more democratic State and is played out in particular ways in hospital settings. Analyses of the South African health care system and its interface with patients in hospitals seldom draw on the work of Foucault, despite its pertinent description of the diffuse and insidious forms of social surveillance (the 'gaze') and processes of 'normalization' brought about in panoptical settings. The gaze has become a metaphor for the processes whereby disciplinary 'technologies', together with the emergence of a normative social science, discipline both the mind and body of the individual, as in my example of a medicalised institutional setting. Transformation from an oppressive State system to a democratic South Africa has impacted in particular ways on the hospital setting. Instead of being subject to the constant surveillance of the gaze of the State or of medicine, there are numerous instances where patients to all intents become 'invisible', and end up beyond its perimeters. In the hospital, as in the heterogeneous South African community, there is a continuous process of adjustment, with patients, services and staff being rotated in an attempt to provide redress and equal access to health services for all. A large, long-standing lack of funds forces the medical staff to make decisions as to who should get access to beds and to optimal care, and this permeates the everyday experience of institutionalisation and care-giving. It also militates against neutral policy objectives, consistent surveillance or a homogenised system of care. PMID- 15351470 TI - Poverty and violence, frustration and inventiveness: hospital ward life in Bangladesh. AB - An ethnographic exploration was done in an orthopaedic ward of a government teaching hospital in Bangladesh to understand the nature of hospital culture in the context of Bangladeshi society at large. Life and work in the ward result in a culture that is simultaneously created by its inhabitants and the conditions in which they are situated. The study shows that biomedicine is a product of particular social conditions and that the hospital reflects features of its society. Behind the injuries and broken limbs in the ward are stories of violence, crime, and intolerance occurring in a society where masses of people fight over limited resources. In the ward people interact in an extremely hierarchical manner. The patients, who are mainly from poor economic backgrounds, remain at the bottom of the hierarchy. Doctors and other staff members are often professionally frustrated. Strikes related to hospital staff's various professional demands hamper the regular flow of work in the ward. Family members are engaged in nursing and provide various kinds of support to their hospitalized relatives. Patients give small bribes to ward boys and cleaners to obtain their day-to-day necessities. Patients joke with each other and mock senior doctors. Thus, they neutralize their powerlessness and drive away the monotony of their stay. Doctors develop 'indigenous' solutions to orthopaedic problems. Instead of using high-tech devices, they employ instruments made of bamboo, bricks, and razor blades. This study shows how medical practice takes shape in an understaffed, under-resourced and poorly financed hospital operating in a low income country. PMID- 15351471 TI - Biomedicine globalized and localized: western medical practices in an outpatient clinic of a Mexican hospital. AB - Following contemporary globalization, biomedicine and western style hospitals have penetrated most corners of the world. We must therefore ask, "How has the diffusion of biomedicine impacted biomedicine's core features of practice cross culturally? How do physicians in different countries make diagnoses, explain etiology and treat patients? To what degree does a physician's cultural understanding shape biomedicine?" Based on extensive fieldwork in a Mexican hospital (Physicians at work, patients in pain. Revised with new preface, Carolina Academic Press, Durham, 2001), this study analyzes the ways in which biomedicine becomes culturally reinterpreted as it moves from one cultural venue to another, and explores the theoretical and practical consequences of this reinterpretation. This analysis illuminates the relationship between biomedicine and the nature of social transformations and refines our understanding of globalization. From a practical perspective, the study is important because a nation's epidemiological profiles are based on statistics drawn from the diagnoses that physicians make. We must not assume that because the same medical nomenclature is used to make the diagnoses, these diagnoses are based on culturally neutral and uniform assessments. PMID- 15351472 TI - Compliance as strategy: the importance of personalised relations in obstetric practice. AB - This paper highlights the importance of personalised relations in institutionalised obstetric care. It seeks to explore the link between objectification and agency, by examining the way in which women find a new subjectivity in motherhood within the walls of the obstetric institution. The paper focuses on obstetric encounters through the lens of labour and intrapartum routine procedures, and argues that when expectant women enter the obstetric institution, a series of relations ensue through their efforts to become connected with the official obstetric system. The hospital organisation and network of relations that originate in it reflect and reproduce a passive role for expectant women and mothers; however, it would be misleading to represent their behaviour as simply "compliant". Expectant mothers value the connectedness with the obstetric system exemplified by personalised patient-practitioner relations because it is a guarantee of safety. They are happy to follow clinicians' instructions, even when this implies significant sacrifice or suffering because they are projected towards fulfilling their parenting desires. Compliance, like resistance then, is here intended to be understood as yet another maternal strategy, and as such it is the most widespread, deployed by women in order to achieve their ideals and desires. PMID- 15351473 TI - Dealing with doubt: making decisions in a neonatal ward in The Netherlands. AB - Neonatology provides intensive care for newborns. Most of the patients in a neonatal ward have been born prematurely. In this article decision-making concerning children born very prematurely (at fewer than 28 weeks, 12 or more weeks too early) is described. Three phases of daily practice are discussed: birth, treatment and referral or death. The article is based on ethnographic research conducted in the neonatal ward of the Amsterdam University Hospital. This ward's policy is to refrain from starting life-prolonging treatment for some children. This is done when staff members consider their chances too small. When life-prolonging treatment is started it is done provisionally and with trial-like features. Staff members want to judge the acceptability of the child's future quality of life. If they think it is unacceptable, they want to stop life prolonging treatment. Parents play an important role in decision-making because their consent is needed for such decisions. More importantly, staff members need parental input about what, for that particular child and those parents, is seen as a good, bad or acceptable quality of future life. Decision-making shows a specific characteristic. Parents are informed about their child in sober terms and because they have considerable opportunities to influence decision-making it is argued that the processes taking place in the ward reflect the Dutch 'negotiation culture'. PMID- 15351474 TI - Doctors and retribution: the hospitalisation of compensation claims in the Highlands of Papua New Guinea. AB - The cultures in the Papua New Guinea Highlands are characterised by a tradition of retribution. Compensation is part of an elaborate system of exchanging gifts, goods and services. Compensation is paid to those who have suffered some kind of loss for which others are held responsible. Such incidents include death or injury caused by fighting, a road accident or domestic violence, theft, rape, gossip, and property damage. Fear of revenge is an important motive for paying compensation. The hospital has become an increasingly important institution for retribution. It provides medical reports to support compensation claims of physical damage in cases involving violence or an accident. Case material, collected by one of the authors who conducted fieldwork in a hospital in the Southern Highlands, shows that the hospital has established itself as an authoritative actor in the local compensation culture. Doctors spend about one afternoon per week writing medical reports for compensation claims. These reports have become an attractive extra source of income for the hospital. The article describes and analyses a number of cases to illustrate the hospital's role in the production and legitimisation of retribution. PMID- 15351475 TI - Privacy, privatization, and the politics of patronage: ethnographic challenges to penetrating the secret world of Middle Eastern, hospital-based in vitro fertilization. AB - In recent years, in vitro fertilization (IVF) has spread around the globe, including to the Middle East. Privacy, privatization, and the politics of patronage are all key issues affecting anthropological research in Middle Eastern hospital-based IVF clinics. IVF-seeking patients generally desire privacy, even total secrecy, when pursuing these treatments, due to cultural issues of stigmatization, particularly regarding male infertility. Thus, ethical issues surrounding the informed consent process are of prime importance. Furthermore, privatization of medical services in the Middle East has left patients--and anthropologists--with few choices other than private IVF clinic settings in which to pursue treatment and research. Both the ethos of patient privacy and medical privatization affect the ability of anthropologists to "penetrate" the secret world of IVF. Permission to conduct ethnography in private hospital IVF clinics may be difficult to obtain without the help of highly motivated physician patrons, who are willing to recruit their private IVF patients for ethnographic interviewing. This article provides a personal account of some of these challenges as faced by a medical anthropologist during a 15-year career of hospital-based IVF research in the Middle East. PMID- 15351476 TI - Social inequalities and disability in older men: prospective findings from the British regional heart study. AB - The independent association of socio-economic position with self-reported disability was assessed. The effect of home and car ownership as additional indices of socio-economic position within occupational social classes was explored. Data from a prospective study of a cohort of 7735 men aged 40-59 years at recruitment and representative of the occupational social class distribution of middle-aged men in Great Britain were used. Men were selected from one general practice in each of 24 towns in England, Wales and Scotland in 1978-1980. The present study concerns 5773 (88.4% of those able to take part) men aged 52-73 years at follow up in 1992 who completed the disability section of a postal questionnaire. A quarter (1453) of men reported disability. Socio-economic position measured as both occupational class (social class I vs. V: age-adjusted OR 5.0, 95% CI 3.4-7.5) and ownership of home and car (both vs. neither: age adjusted OR 2.8, 95% CI 2.3-3.4) showed a graded relationship with likelihood of reporting disability in 1992. Within all social class groups, those owning both home and car had a lower risk of disability than those who owned neither, even after adjustment for a wide range of risk factors. Men from manual occupations were more likely than those in non-manual occupations to report disability on developing chronic diseases. The relationship between socio-economic position and severe, but not milder, disability appeared to be independent of disease status. Socio-economic position is a strong predictor of disability in later life independent of a wide range of lifestyle factors and presence of diagnosed disease. The likelihood of reporting disability between and within social class groups is influenced by material wealth. PMID- 15351477 TI - Gender differences in factors affecting use of health services: an analysis of a community study of middle-aged and older Australians. AB - Research on patterns of self-rated health and health service use suggests that women report having poorer health than men, and that, after controlling for health measures, women are more likely to obtain formal health care. Proposed reasons for these differences have included that women's self-rated health is more strongly influenced by psychosocial factors or negative affect and that women are likely to obtain services when at better levels of self-rated health, compared with men. Our study explored gender differences in the effects of non health attributes on decisions to obtain primary medical services for an Australian community-based sample of 4140 adults from two age groups: 40-44 years and 60-64 years. Participants provided information on measures of physical and mental health, and on predisposing and enabling factors that could affect their levels of health service use. Information on visits made to general practitioners (GPs) in a 6-month period was obtained from the national insurer. We found that men and women who obtained no GP services reported comparable levels of physical and mental health and that, for both men and women, measures of health needs were most strongly associated with their obtaining care. After controlling for measures of mental and physical health and enabling factors, we found that non health factors that could predispose an individual to obtain care had greater impact on men's but not women's decisions to obtain any GP services. Our findings do not support the hypothesis that in choosing to obtain medical care, women are more strongly influenced by non-health factors compared with men. PMID- 15351478 TI - Childhood IQ and cardiovascular disease in adulthood: prospective observational study linking the Scottish Mental Survey 1932 and the Midspan studies. AB - This study investigated the influence of childhood IQ on the relationships between risk factors and cardiovascular disease (CVD), coronary heart disease (CHD) and stroke in adulthood. Participants were from the Midspan prospective cohort studies which were conducted on adults in Scotland in the 1970s. Data on risk factors were collected from a questionnaire and at a screening examination, and participants were followed up for 25 years for hospital admissions and mortality. 938 Midspan participants were successfully matched with their age 11 IQ from the Scottish Mental Survey 1932, in which 1921-born children attending schools in Scotland took a cognitive ability test. Childhood IQ was negatively correlated with diastolic and systolic blood pressure, and positively correlated with height and respiratory function in adulthood. For each of CVD, CHD and stroke, defined as either a hospital admission or death, there was an increased relative rate per standard deviation decrease (15 points) in childhood IQ of 1.11 (95% confidence interval 1.01-1.23), 1.16 (1.03-1.32) and 1.10 (0.88-1.36), respectively. With events divided into those first occurring before and those first occurring after the age of 65, the relationships between childhood IQ and CVD, CHD and stroke were only seen before age 65 and not after age 65. Blood pressure, height, respiratory function and smoking were associated with CVD events. Relationships were stronger in the early compared to the later period for smoking and FEV1, and stronger in the later compared to the earlier period for blood pressure. Adjustment for childhood IQ had small attenuating effects on the risk factor-CVD relationship before age 65 and no effects after age 65. Adjustment for risk factors attenuated the childhood IQ-CVD relationship by a small amount before age 65. Childhood IQ was associated with CVD risk factors and events and can be considered an important new risk factor. PMID- 15351479 TI - The association of personal and neighborhood socioeconomic indicators with subclinical cardiovascular disease in an elderly cohort. The cardiovascular health study. AB - There has been recent interest in determining whether neighborhood characteristics are related to the cardiovascular health of residents. However, there are no data regarding the relationship between neighborhood socioeconomic status (SES) and prevalence of subclinical cardiovascular disease (CVD) in the elderly. We related personal SES (education, income, and occupation type) and neighborhood socioeconomic characteristics (a block-group score summing six variables reflecting neighborhood income and wealth, education, and occupation) to the prevalence of subclinical CVD (asymptomatic peripheral vascular disease or carotid atherosclerosis, electrocardiogram or echocardiogram abnormalities, and/or positive responses to Rose Questionnaire claudication or angina pectoris) among 3545 persons aged 65 and over, without prevalent CVD, in the Cardiovascular Health Study. Sixty percent of participants had at least one indicator of subclinical disease. Compared to those without, those with subclinical disease had significantly lower education, income, and neighborhood scores and were more likely to have blue-collar jobs. After adjustment for age, gender, and race, those in the lowest SES groups had increased prevalence of subclinical disease compared with those in the highest SES groups (OR = 1.50; 95% CI 1.21, 1.86 for income; OR = 1.41; 95% CI 1.18, 1.69 for education; OR = 1.39; 95% CI 1.16, 1.67 for block-group score). Those reporting a blue-collar lifetime occupation had greater prevalence of subclinical disease relative to those reporting a white collar occupation (OR = 1.29; 95% CI 1.02-1.59). After adjustment for behavioral and biomedical risk factors, all of these associations were reduced. Neighborhood score tended to remain inversely associated with subclinical disease after adjustment for personal socioeconomic indicators but associations were not statistically significant. Personal income and blue-collar occupation remained significantly associated with subclinical disease after simultaneous adjustment for neighborhood score and education. Personal and neighborhood socioeconomic indicators were associated with subclinical disease prevalence in this elderly cohort. These relationships were reduced after controlling for traditional CVD risk factors. PMID- 15351480 TI - Clarifying the relationships between health and residential mobility. AB - Health-selective migration within countries has been implicated as one of the mechanisms by which spatial disadvantage is created and maintained. However, there is conflicting evidence on the nature of the relationship between health and mobility, caused in part by diverse definitions, and age and sex differences. This paper uses the first two waves of data for the middle-aged cohort (aged 45 50 in 1996) of the Australian Longitudinal Study on Women's Health to investigate the relationship between four sets of health variables with subsequent local moves (within the same postcode), longer distance moves (between postcodes) and inter-regional migration from rural and remote areas 'up' the urban hierarchy. After adjusting for socio-economic and marital status, short and longer distance mobility among these middle-aged Australian women was positively associated with long-term and chronic poor health and being a smoker. Moves between postcodes and rural-to-urban migration were positively associated with multiple recent visits to a medical specialist. Our findings are consistent with UK and US studies that have found mobility to be more strongly associated with poor health than good health in mature adults. As the population ages, the health of receiving areas may be adversely affected by relatively unhealthy in-migrants seeking amenities not provided in their former place of residence. PMID- 15351481 TI - The elderly and AIDS: coping with the impact of adult death in Tanzania. AB - The elderly are one of the key groups of survivors who may be adversely affected by the death of prime-age adults from AIDS. We use a longitudinal survey of households from Northwestern Tanzania in 1991-94 to compare the activities and welfare of the elderly in households before and after the death of a prime-age adult with that of the elderly in households that did not have an adult death. The elderly in households that had an adult death were more educated, more likely to engage in wage employment and spent less time farming than the elderly in households that did not have a death during the survey. Time spent by the elderly in household chores rose following an adult death and participation in wage employment fell; there was no evidence of increased participation in farm work. The physical well-being of the elderly, as measured by body mass index (BMI), was lower prior to an adult death and higher afterward. Finally, the elderly with the lowest BMI are those in poor households that did not have an adult death during the survey. Thus, policymakers should be concerned about the adverse impacts of adult deaths on the physical well-being of the elderly-primarily during the period of illness prior to a prime-age adult death-but they should also focus on the larger groups of poor elderly with much lower physical health status. PMID- 15351482 TI - Organ transplantation in Singapore: history, problems, and policies. AB - The article explores the rules and criteria used for the allocation of donor organs in Singapore. Organs are extremely scarce, so hard decisions have to be made about who receives them. Based on an analysis of documents and interviews done with local program directors, both the admission to a waiting list for transplant and the final recipient selection are covered. The observed practices appear remarkably similar to those predominant in several Western countries until recently, when they had to be modified there following public criticism. It remains to be seen whether Singapore can sustain its much stricter standards in the future. PMID- 15351483 TI - Heparin binding activity of orf virus F1L protein. AB - The orf virus is the type species of the Parapoxvirus genus and is the causative agent of contagious echtyma, a debilitating skin disease of sheep and goats, which can also affect man. The virus exhibits a restricted host range, even if it has been shown to bind to a wide range of tissues of non-permissive species. This ability is an argument for its potential use as an expression vector. Since most mammalian cell types express heparan sulfate (HS) surface receptors, we assumed that HS could serve as receptors to mediate orf virus binding. In this study, we showed that orf virus is inhibited by the addition of soluble heparin in cell cultures. Affinity chomatography using heparin agarose demonstrated that orf virus F1L is the major heparin binding protein. Furthermore, the recombinant F1L protein was visualised on the cell surface by confocal microscopy, and rabbits immunised with recombinant F1L protein produced virus neutralising antibodies. These results confirm that the F1L immunodominant protein is also involved in virus binding to cells as for the vaccinia homologue H3L protein. Heparin also inhibited the binding of the F1L protein to cells showing that this protein has a role in the early stages of infection. PMID- 15351484 TI - Heliocoverpa armigera single nucleocapsid nucleopolyhedrovirus induces Hz-AM1 cell cycle arrest at the G2 phase with accumulation of cyclin B1. AB - The cell cycle phase distributions of Hz-AM1 cells grown in monolayer culture were G1 = 49.7 +/- 3.3%, S = 22.7 +/- 3.8% and G2/M = 27.8 +/- 4.2% without >4N DNA content. The culture doubling time was about 40 h and the duration of the G1, S and G2/M phases was estimated to be 10, 14 and 16 h, respectively. HaSNPV infection of Hz-AM1 cells resulted in both unsynchronized and synchronized G1 phase arrest at G2/M phase. HaSNPV infection also resulted in the appearance of more than 4N DNA content, which accumulated to the highest levels 72 h post infection. We also found that the expression level of cyclin B1 increased significantly after 16 h post-infection, while cyclin A did not show any change. This observation supports the Hz-AM1-infected arrest at the G2/M phage. Cytoplasm location of cyclin B1 indicated that the Hz-AM1 cell cycle arrest was at the G2 phase. PMID- 15351485 TI - Characterization of protein-protein interactions between the nucleocapsid protein and membrane protein of the SARS coronavirus. AB - The human coronavirus, associated with severe acute respiratory syndrome (SARS CoV), was identified and molecularly characterized in 2003. Sequence analysis of the virus indicates that there is only 20% amino acid (aa) identity with known coronaviruses. Previous studies indicate that protein-protein interactions amongst various coronavirus proteins are critical for viral assembly. Yet, little sequence homology between the newly identified SARS-CoV and those previously studied coronaviruses suggests that determination of protein-protein interaction and identification of amino acid sequences, responsible for such interaction in SARS-CoV, are necessary for the elucidation of the molecular mechanism of SARS CoV replication and rationalization of anti-SARS therapeutic intervention. In this study, we employed mammalian two-hybrid system to investigate possible interactions between SARS-CoV nucleocapsid (N) and the membrane (M) proteins. We found that interaction of the N and M proteins takes place in vivo and identified that a stretch of amino acids (168-208) in the N protein may be critical for such protein-protein interactions. Importantly, the same region has been found to be required for multimerization of the N protein (He et al., 2004) suggesting this region may be crucial in maintaining correct conformation of the N protein for self-interaction and interaction with the M protein. PMID- 15351486 TI - Porcine adenoviral vectors evade preexisting humoral immunity to adenoviruses and efficiently infect both human and murine cells in culture. AB - Preexisting immunity against human adenoviruses (HAd) limits the efficiency of transduction of HAd vectors in humans. In addition, development of a vector specific immune response after the first inoculation with a HAd vector further lowers vector uptake following readministration. We investigated the usefulness of porcine adenovirus serotype 3 (PAd3)-based vectors as a supplement to HAd vectors. Here we demonstrate that preexisting HAd-specific neutralizing antibodies in humans do not cross-neutralize PAd3. In order to generate E1A deleted PAd3 vectors, an E1-complementing cell line of porcine origin was produced. E1A-deleted PAd3 vector expressing green fluorescent protein; GFP (PAd GFP) and E1-deleted HAd5 vector expressing GFP (HAd-GFP) transduced human cell lines with comparable efficiencies. Both of these vectors efficiently transduced murine MT1A2 breast cancer cell line, while PAd-GFP transduced murine NIH 3T3 fibroblast cell line significantly better (P < 0.05) than HAd-GFP. These results suggest that PAd3 vectors would be promising supplement to HAd vectors as a delivery vehicle for recombinant vaccines and gene therapy applications. PMID- 15351487 TI - Molecular epidemiology of respiratory syncytial virus (RSV) of group A in Stockholm, Sweden, between 1965 and 2003. AB - The epidemiology of respiratory syncytial virus (RSV) group A was followed by nucleotide sequencing of the variable parts of the glycoprotein (G) gene. The amino acid sequences of an aminoterminal (A-terminal, amino acids 90-132) and carboxyterminal (C-terminal, amino acids 262-298) portion of the G protein in 47 virus strains, collected in Stockholm, between 1965 and 2004, were determined. In phylogenetic analysis jointly with previously described genotypes (GA1 to GA7 and SAA1), 33 virus strains (isolated between 1991 and 2004) belonged to genotype GA5, seven to GA2, three to genotype GA1 (isolated before 1991), one to genotype GA4 (isolated in 1982) and three to genotype GA7 (isolated in 1993 and 2001). Genotype GA5 was predominant in four epidemics, between 2000/2001 and 2003/2004. Little or no variation with time of the C-terminal amino acid sequence of the G protein was found when the virus strains were compared within their own genotype. Identical and nearly identical nucleotide sequences were found between strains isolated more than 10 (GA5) and 25 (GA2) years apart. The A-terminal part of they G protein of genotype GA2 was highly conserved. In contrast, the A-terminal part of the G protein of genotype GA5 exhibited a pronounced variation in its amino acid sequence over time. PMID- 15351488 TI - Complete gene sequence of a human Puumala hantavirus isolate, Puumala Umea/hu: sequence comparison and characterisation of encoded gene products. AB - Puumala virus is a member of the hantavirus genus in the Bunyaviridae family, and the major causative agent of haemorrhagic fever with renal syndrome in Europe. This study was conducted with a human Puumala virus isolate (PUUV Umea/hu), and contains the determination of the first complete PUUV sequence from a human source. When the relationship to other Puumala viruses was analysed, a possible RNA segment exchange between two local strains of PUUV was noticed. Furthermore, the coding regions of PUUV Umea/hu S- and M-segments were cloned, and a large set of gene products were expressed in mammalian cells. In addition, postulated N- and O-linked glycosylation sites in the two envelope proteins (Gn and Gc) were investigated individually by site-directed mutagenesis followed by gel-shift analysis. Our data demonstrate that N-linked glycosylation occurs at three sites in Gn (N142, N357 and N409), and at one site in Gc (N937). Also, one possible O glycosylation site was identified in Gc (T985). We conclude that the diversity between different Puumala virus isolates is high, and consequently characterization of local PUUV isolates is important for clinical diagnostic work. Finally, the obtained results concerning the encoded gene products are of great importance for the design of new vaccines. PMID- 15351489 TI - The conserved amino-terminal region (amino acids 1-20) of the hepatitis B virus X protein shows a transrepression function. AB - The X protein of hepatitis B virus or HBx is a multifunctional regulatory protein that carries the fame of a promiscuous transactivator. Although, the N-terminal 'A' region of HBx (amino acids 1-20) is the most conserved region among mammalian hepadnavirus genomes, it has been found to be dispensable for transactivation function [Proc. Natl. Acad. Sci. U.S.A. 93, 1996, 5647]. To elucidate its biological role, DNA sequence corresponding to the A region of X gene was amplified by polymerase chain reaction and cloned as a 72 base pair HBx mutant X17. In order to augment the intracellular biochemical stability of the expressed protein, the monomeric X17 was multimerized and 2-10 units long tandem repeats of the A region (X17-n) were cloned in a mammalian expression vector. Expression of the X17 constructs was confirmed by in vitro transcription and translation, as well as by RT-PCR after transfection in hepatoma cells. The function of X17 was investigated using the chloramphenicol acetyl transferase reporter constructs of viral (RSV-LTR, HIV1-LTR and HBx) and cellular gene promoters (c-Jun and epidermal growth receptor). Not only did the X17 multimers inhibit the HBx mediated transactivation of all the reporter genes, but also their basal activities. The inhibition was dependent on the amount of X17 plasmid transfected in cells as well as on the number of repeat units present in the X17 expression vectors. Further, the X17-related inhibition of transactivation was not a cytotoxic effect. Thus, our data suggests that the N-terminal 'A' domain of HBx has a negative regulatory function. PMID- 15351490 TI - Phylogenetic relationship of porcine endogenous retrovirus (PERV) in Chinese pigs with some type C retroviruses. AB - PCR amplification of proviral DNA extracted from peripheral blood lymphocytes of three Chinese pigs (Banna minipig inbreed (BMI), Wu-Zhi-Shan pig (WZSP) and Neijiang pig (NJP)), using primers corresponding to highly conserved regions of reverse transcriptase (RT) of pol gene and nucleocapsid sequence of gag gene. PCR products were then extracted and cloned into pGEM-T vector. Phylogenetic analysis of the nucleotide sequences of PERV-BMI, PERV-WZSP and PERV-WZSP revealed that they were of retroviral origin. Phylogenetic trees were constructed from the translated amino acids of PERVs and other type C retrovirus, as well as lentivirus of GenBank. The research demonstrated that PERVs of Chinese pigs and other PERVs were closely related to other pathogenic type C retroviruses. From the gag analysis, a novel subgroup of PERV was identified and this novel sequence described in this report would allow such investigation to be actively pursued. PMID- 15351491 TI - Inhibitory effects of cystatins on proteolytic activities of the Plum pox potyvirus cysteine proteinases. AB - In an effort to develop new antiviral strategies effective against potyviruses, several cystatins were evaluated for their ability to inhibit the cysteine proteinases of Plum pox potyvirus (PPV) using in vitro proteolytic assays. The following cystatins were purified as GST fusion proteins and shown to be active against papain:oryzacystatins I and II (OCI and OCII), corn cystatin II (CCII), human stefin A (HSA), the domain 8 of tomato multicystatin (TMC-8) and a large 24kDa tomato cystatin (LTCyst). These cystatins did not inhibit the activity of purified recombinant PPV NIa proteinase, a serine-like cysteine proteinases related to the 3C proteinases of picornaviruses and to chymotrypsin. The cystatins were shown to inhibit slightly the activity of the PPV HC-Pro proteinase with CCII being the best inhibitor. However a large excess of the cystatins was required to observe any inhibition. Based on these results and on the documented pleiotropic effects of cystatins on the metabolism of plants, we conclude that they are not the best candidates for antiviral strategies targeted to viral cysteine proteinases. The availability of soluble active recombinant PPV NIa proteinase will be instrumental for the selection of other proteinase inhibitors with increased affinity and specificity for this proteinase. PMID- 15351492 TI - Protective efficacy of intranasal cold-adapted influenza A/New Caledonia/20/99 (H1N1) vaccines comprised of egg- or cell culture-derived reassortants. AB - Live, cold-adapted, temperature-sensitive (ca/ts) Russian influenza A vaccines are prepared in eggs by a 6:2 gene reassortment of the ca/ts donor strain A/Leningrad/134/17/57 (H2N2) (Len/17) with a current wild-type (wt) influenza A strain contributing hemagglutinin (HA) and neuraminidase (NA) genes. However, egg derived reassortant vaccines are potentially more problematic to manufacture in large quantities than vaccines from cell-based procedures. To compare egg- and cell culture-derived reassortant vaccines, we prepared in Madin Darby canine kidney (MDCK) cells two cloned, ca/ts reassortants (25M/1, 39E/2) derived from Len/17 and a wt reference strain A/New Caledonia/20/99 (H1N1) (NC/wt). Both 25M/1 and 39E/2 reassortants preserved the ca/ts phenotype and mutations described for internal genes of the A/Len/17 parent. When compared to a commercial, egg-derived ca/ts Russian A/17/NC/99/145 (H1N1) New Caledonia vaccine (NC/145), the MDCK derived reassortant 39E/2 vaccine conferred similar levels of protection in ferrets challenged i.n. with 7 x 10(10) pfu of NC/wt. In a dose-ranging study, the protective vaccine dose for 50% of ferrets (PD50) was less than 1.2 x 10(4) pfu for the 25M/1 vaccine derived by recombination and amplification in MDCK cells. Clonal isolates of ca/ts influenza A/New Caledonia/20/99 (H1N1) obtained by recombination and amplification entirely in MDCK cells can be highly protective i.n. vaccines. PMID- 15351493 TI - Sequence analysis, viral rescue from infectious clones and generation of recombinant virions of the avian adeno-associated virus. AB - Aiming at the generation of a viral-vectored system for gene delivery and vaccination in poultry, the entire genomes of the VR-865 and DA-1 strains of the avian adeno-associated virus have been cloned and sequenced. Sequence analysis of the clones showed that the genomic distribution of the structural and non structural protein-coding genes of these viruses is conserved and in agreement with what has been previously described for the primate adeno-associated viruses. Amino acid differences between the avian adeno-associated viruses and the primate adeno-associated viruses are more evident in the genes that code for the non structural (Rep) proteins of the virus, while the Cap region amino acid sequence was found to be more conserved. Since all the regulatory and coding sequences of the virus were present in the plasmids obtained, complete infectious viral particles were rescued from these clones, and these rescued viral populations were amplified by co-infecting primary embryo liver cells with the rescued virus and the CELO strain of the avian adenovirus type 1. As a proof of concept of the validity of this system for the purpose of gene delivery, recombinant viruses encoding for the LacZ gene as a reporter system were also generated. These recombinant viruses were used to express beta galactosidase activity in primary chicken embryo cell cultures. PMID- 15351494 TI - Close proximity of the MPMV CTE to the polyadenylation sequences is important for efficient function in the subgenomic context. AB - The constitutive transport element (CTE) of Mason-Pfizer monkey virus (MPMV) is a short cis-acting sequence element critical for virus gene expression. Analogous to the Rev/Rev Responsive Element (RRE) of primate lentiviruses, CTE allows the nucleocytoplasmic transport of unspliced viral mRNAs. In fact, CTE can functionally replace Rev/RRE in the genomic context and has been used successfully in the expression of viral and cellular genes from expression vectors as well. However, unlike RRE, CTE accomplishes this by interacting with cellular factors, making CTE function independent of co-expressed trans factors. Thus, CTE has proven to be a valuable tool in the expression of heterologous genes. Our previous studies have shown that close proximity of CTE to the polyadenylation sequences is important for CTE function in the genomic context. However, it is controversial whether CTE needs to be located spatially close to the polyadenylation sequences in the subgenomic context. Since CTE is being frequently used in expression vectors, we investigated the position dependency of CTE in the heterologous, subgenomic background using both genetic and structural analyses. Our results reveal that similar to the genomic situation, close proximity of CTE to the polyadenylation sequences is important for its function in the heterologous subgenomic context. PMID- 15351495 TI - Age and experience-related improvements in gap detection in the rat. AB - The ability to accurately process brief, successive acoustic signals rapidly presented to the central nervous system is believed to underlie successful language development. The limits of temporal resolution of the auditory system, often assessed using gap detection tasks, has been widely studied in relation to developing and decoding speech. In the present study, a reflex modification paradigm was used to investigate potential shifts in gap detection thresholds in rats across development, with test sessions beginning on postnatal day (P) 15, P35 and P64. We found that thresholds decreased over the course of development. These thresholds were determined to lie between 10 and 20 ms for the P15 and P35 groups, and between 5 and 10 ms for the P64 group. Moreover, we observed improvements in gap detection thresholds in all age groups over 5 days of testing, including the youngest age group (P15). These later results suggest that experience-dependent plasticity mechanisms at the level of sensory processing are operational and observable both very early in development, and also in adult animals. The present findings also demonstrate maturational improvements in silent gap detection using a pre-pulse inhibition paradigm. PMID- 15351496 TI - Impaired gap detection in juvenile microgyric rats. AB - Previous research with adult animal models links the presence of cortical neuromigrational anomalies (i.e., microgyria similar to that found in brains of dyslexics) with rapid auditory processing (RAP) impairments. RAP impairments are in turn found in children with specific language impairment (SLI) and also in individuals with dyslexia. Gap detection, a simple measure of auditory temporal acuity, appears to be impaired in children with SLI but not in dyslexic adults, even though both groups exhibit impaired processing on more complex, rapid auditory tasks. In the current study, juvenile rats with bilateral microgyria, but not their adult counterparts, exhibited impaired detection of short duration silent gaps in white noise when compared to age-matched sham littermates. Results lend further support to: (1) an association between neuromigrational anomalies and RAP impairments; and (2) the validity of an animal model of RAP impairments associated with language disturbances in humans. Current results also support the view that auditory processing disturbances associated with cortical malformations may be evident early in development at a relatively "low" level (e.g., simple gap detection), but may require "higher-order" auditory discrimination tasks (e.g., tone sequences, phonemic discriminations) to be elicited later in life. PMID- 15351497 TI - Developmental changes of GABAergic synapses formed between primary cultured cortical neurons. AB - The characteristics of functional changes of GABAergic synapses between cultured rat cortical neurons were observed by monitoring intracellular calcium level ([Ca2+]in) during development in vitro. After 5 days in vitro (DIV), cultured cortical neurons spontaneously exhibited synchronous oscillatory changes in [Ca2+]in, which were derived from synaptic activity. Exposure to bicuculline, antagonist of gamma-aminobutyric acid (GABA)(A) receptors, caused a marked decrease in the frequency of [Ca2+]in oscillations at 7-20 DIV. Although the frequency of spontaneous oscillations increased during this culture period, the ratio of the decrease in the frequency following bicuculline treatment did not significantly change. Thereafter, to investigate the detailed morphological changes of GABAergic synapses during development in vitro, the cultured neurons were immunostained with antibodies to glutamic acid decarboxylase (GAD), synaptophysin and GABA(A) receptor and were observed under a confocal laser microscope. Most of the GAD-positive puncta colocalized with synaptophysin positive puncta and were opposed to GABA(A) receptor-positive structures. The images of GAD-positive puncta were reconstructed from the confocal three dimensional data to analyze their number, volume, and surface area. The number of these puncta increased with culture time at 7-20 DIV. Although the volume of individual GAD-positive puncta did not significantly change, the surface area decreased in a time-dependent manner over the culture period. This system that we developed enabled us to investigate in detail the morphological and functional changes of GABAergic synapses during neuronal development. PMID- 15351498 TI - Developmental expression of RFamide-related peptides in the rat central nervous system. AB - RFamide-related peptides (RFRP-1 and RFRP-3) have been recently identified in mammals and considered to play significant functional roles in the rat brain. In this study, we report the developmental expression of RFRP mRNA and its immunoreactive neuronal cells and fibers in the rat brain. The RFRP mRNA was expressed in the brain from embryonic day 15 (E15) according to reverse transcription-polymerase chain reaction analysis. We first detected RFRP mRNA expressing neurons in the caudal portion of the hypothalamus at E16 by in situ hybridization analysis. Immunohistochemical analysis showed that RFRP-3 or RFRP-1 immunoreactive neuronal cell bodies were first detected at E16 or E17, respectively. Double-labeling fluorescent immunohistochemical analysis showed that neurons containing both RFRP-1 immunoreactivity (ir) and RFRP-3-ir were detected from E18. We also detected RFRP-1 immunoreactive nerve fiber processes in the forebrain, hypothalamus, thalamus, midbrain, pons and medulla oblongata at prenatal day and the distribution of RFRP-1 immunoreactive nerve fibers in postnatal day 0 (P0) were almost coincident with that in adult. However, localization of RFRP-3 immunoreactive nerve fibers was limited around the RFRP-3 immunoreactive neuronal cell bodies during prenatal days. The distribution of RFRP-3 immunoreactive nerve fibers was first detected in the above areas at P0. The nerve fibers containing only RFRP-3-ir in the thalamus or spinal cord were first appeared at P21 or P28. Our results show that RFRP mRNA was expressed during the neonates and the distribution of RFRP-1 or RFRP-3 immunoreactive nerve fibers would be distinctly regulated in the developing rat brain. PMID- 15351499 TI - Co-localization of fukutin and alpha-dystroglycan in the mouse central nervous system. AB - Hypoglycosylation of alpha-dystroglycan (alpha-DG) has been identified in several human diseases associated with muscular dystrophy and brain malformations, including Fukuyama-type congenital muscular dystrophy (FCMD) caused by mutations in the fukutin gene. Although disruption of the intra-extra membrane linkage in the sarcolemma via the dystroglycan (DG) has been hypothesized as a possible underlying mechanism, little is known about the pathogenesis of brain anomalies in these conditions. In this study, we examined the patterns of expression of fukutin and alpha-DG in developing and adult mouse brains. Antisera against fukutin and alpha-DG identified neurons of the fetal cerebral and cerebellar cortex and the subpial pontine migratory stream. In adult mice, fukutin and alpha DG were extensively co-expressed in neurons of the cerebral and cerebellar cortex, hippocampus, basal ganglia and olfactory bulb, as well as in the pontine nucleus and the cranial nerve nuclei. These results support the hypothesis that fukutin is involved in the glycosylation process of alpha-DG and that a defect in this process plays an essential role in the pathogenesis of FCMD. Further research into the physiological function of alpha-DG in migrating and mature neurons is required. PMID- 15351500 TI - Expression of plasma membrane Ca2+ ATPase family members and associated synaptic proteins in acute and cultured organotypic hippocampal slices from rat. AB - Plasma membrane Ca2+ ATPases (PMCAs) are critical regulators of intracellular Ca2+ concentration ([Ca2+]i). Specific isoforms have also been demonstrated to interact and co-localise with members of the synapse-associated protein (SAP) family in hippocampal dendritic spines. Presently, only indirect evidence of changes in PMCA protein expression during postnatal development exists, therefore we chose to examine the postnatal developmental protein expression patterns of PMCAs 1-4 and the SAP proteins SAP102 and PSD95. Using Western blotting analysis, we compared the postnatal expression in the in vivo hippocampus to the expression within in vitro organotypic hippocampal slice cultures; a valid model of the developing hippocampus. All PMCA and SAP family members studied showed a marked increase in protein expression levels throughout the postnatal time course both in vivo and in vitro. SAP102 and the ubiquitously expressed PMCAs 1 and 4 followed a similar time course of expression within the in vivo and in vitro preparations. In contrast, the neurone-specific PMCA isoforms 2 and 3 and PSD95 displayed slight differences in early postnatal development. However, and most importantly, their expression > or = 14 days in vitro (DIV) was similar to that in vivo. The results of this study demonstrate that postnatal expression of all PMCAs, SAP102 and PSD95 is similar in both the in vivo hippocampus and the in vitro organotypic hippocampal slice culture. Our results support the use of organotypic hippocampal slice cultures for future investigations of the importance of PMCAs for neuronal Ca2+ handling and SAP family member interactions. PMID- 15351501 TI - Long-term effects of seizures in neonatal rats on spatial learning ability and N methyl-D-aspartate receptor expression in the brain. AB - For the purpose of investigating the long-term effects of seizures in neonatal rats on spatial learning ability and N-methyl-D-aspartate (NMDA) receptor expression in adult rat brain, a seizure was induced by inhalant flurothyl daily in neonatal Wistar rats from postnatal day 6 (P6). The authors assigned six rats each averagely into the single-seizure group, the recurrent-seizure group (seizures induced in six consecutive days), and the control group. During P60 to P65, the rats were tested for spatial learning ability with the Morris water maze task. On P75, the authors examined protein expression of the NMDA receptor (NR) subunits, NR1, 2A, 2B, 2C, and 2D, in the cerebral cortex and hippocampus by Western blotting analysis. On P65, the escape latencies from the water maze of the rats in the recurrent-seizure group were significantly longer than those of the control rats, but there was no difference between the single-seizure group and the control group. NR subunit expression in the cerebral cortex and hippocampus of the rats with single seizure was similar to those in the control rats. Compared with the control rats, the protein expressions of NR1, NR2A and NR2B in the cerebral cortex and NR2A in the hippocampus of the recurrent-seizure group was significantly decreased, but NR2C protein expression in the cerebral cortex and hippocampus significantly increased. Recurrent seizures induced in neonatal rats might cause long-term spatial learning ability deficit and modify NR expression in the cerebral cortex and hippocampus of adult rats. The results suggest that abnormal NR expression might play an important role in long-term spatial learning ability deficit induced by recurrent seizures in early life. PMID- 15351502 TI - Neuropathology of seizures in the immature rabbit. AB - Acute morphologic changes of brain due to chemically induced seizures are studied in developing rabbits. Accordingly, rabbits of postnatal days 6 and 7 (p6-7) and p10-12 are injected with a single dose of 1-6 mg/kg kainic acid (KA) intraperitoneally (i.p.) or injected with a single dose of 200-300 mg/kg pilocarpine subcutaneously (s.c.). Many animals developed seizures of varying severity and length. Histologic examination of brain 2 days following injection showed that KA-induced seizures did not cause neuronal death. Pilocarpine-induced seizures resulted in neuronal death mainly involving the CA1 region of hippocampus. In the p6-7 group, only a small number of brains were involved, lesions were mild and limited to CA1. In the p10-12 group, majority of the brains were damaged, lesions were relatively severe, and in some brains extended beyond the CA1 region involving the subiculum, CA3, cortex, and amygdala. Measurements of physiologic parameters indicate that these changes were not secondary to hypoxemia during seizures. However, there was hypotension and hyperthermia, both of which may contribute to brain damage during seizures. The findings suggest that pilocarpine-induced seizures during the second postnatal week in rabbits is a useful model to study the morphologic changes of brain due to seizure in the developing animal and also to assess the systemic physiologic alterations during seizures. PMID- 15351503 TI - Early postnatal Purkinje cells from staggerer mice undergo aberrant development in vitro with characteristic morphologic and gene expression abnormalities. AB - The staggerer (sg) mutant mouse shows an early block in Purkinje cell (PC) development due to a truncation of the transcription factor retinoic acid-related orphan receptor alpha (ROR alpha). Cerebellar cultures from neonatal sg mice recapitulate many in vivo postnatal changes in PCs and allow for detailed observation of sequential dendritic outgrowth and gene expression changes. In addition, this in vitro system provides a tractable basis for ROR alpha replacement studies. PMID- 15351504 TI - High-level expression of functional chemokine receptor CXCR4 on human neural precursor cells. AB - Neural precursor cells (NPCs) are self-renewing, multipotent progenitors that give rise to neurons, astrocytes and oligodendrocytes in the central nervous system (CNS). Fetal NPCs have attracted attention for their potential use in studying normal CNS development. Several studies of rodent neural progenitors have suggested that chemokines and their receptors are involved in directing NPC migration during CNS development. In this study, we established a consistent system to culture human NPCs and examined the expression of chemokine receptors on these cells. NPCs were found to express the markers nestin and CD133 and to differentiate into neurons, astrocytes and oligodendrocytes at the clonal level. Flow cytometry and RNase protection assay (RPA) indicated that NPCs express high levels of CXCR4 and low levels of several other chemokine receptors. When examined using a chemotaxis assay, NPCs were able to respond to CXCL12/SDF 1alpha, a ligand of CXCR4. Treatment with anti-CXCR4 antibody or HIV-1 gp120 abolished the migratory response of NPCs towards CXCL12/SDF-1alpha. These findings suggest that CXCR4 may play a significant role in directing NPC migration during CNS development. PMID- 15351505 TI - Dose-dependent effects of methylmercury administered during neonatal brain spurt in rats. AB - Rapid brain growth occurs primarily during the third trimester in humans, whereas in rats it occurs after parturition. Therefore, we hypothesized that the effects of methylmercury (MeHg) on the postnatal developing rat nervous system may help in understanding the neurotoxicity on the human fetal brain when the brain is most vulnerable. In the present experiment, the dose-response effects of MeHg treatment during the postnatal developing phase in rats were studied. Male Wistar rats were orally administered 0, 1, 3, and 5 mg/kg/day methylmercury chloride (MMC), respectively, on postnatal day 1 and for 30 consecutive days. The body weight decline began from day 25 and typical symptoms, such as hind-limb crossing and ataxia, were observed in rats treated with 5 mg/kg/day MMC. The weight loss and typical symptoms were not observed in rats treated with 1 and 3 mg/kg/day. Mercury (Hg) concentrations in the brain were 2.6, 4.5, and 9.6 microg/g in the rats treated with 1, 3, and 5 mg/kg/day, respectively, on the day after the final MMC treatment. At 5 to 6 weeks of age, dose-dependent deficits of motor coordination in the rotarod test and learning disability in the passive avoidance response test were observed. Histopathological examination of a proportion of the MeHg-treated rats revealed widespread neuronal degeneration manifested by neuron loss and astrocytosis in the cerebral cortex, striatum, and cerebellum, where severity of the lesions seemed to increase in proportion to the administered dose of MMC. These findings using neonatal rats will be useful for better understanding of the effects of MeHg in the developing human brain during gestation. PMID- 15351506 TI - Prenatal exposure of Long-Evans rats to 17alpha-ethinylestradiol modifies neither latent inhibition nor prepulse inhibition of the startle reflex but elicits minor deficits in exploratory behavior. AB - Prenatal administration of synthetic estrogens in humans as well as lower mammals was reported to alter behavior in adulthood. The alterations remain to be characterized according to specific pathophysiological hypotheses. In this study, three common behavioral models of schizophrenia were tested, i.e., latent inhibition (LI), prepulse inhibition of the startle response (PPI) and hyperlocomotion under amphetamine. Female Long-Evans rats were injected i.p. with a solution of 17alpha-ethinylestradiol (15 microg kg(-1)) everyday from day 9 to 14 of pregnancy, and behavioral characteristics of their offspring, raised by Wistar foster mothers, were compared to those of rats born from dams injected with the vehicle only, over the same gestation period. LI was tested in a conditioned taste aversion and a conditioned passive avoidance paradigm followed by a parametric study of PPI and an evaluation of locomotion in an open field under saline or amphetamine (1.5 mg kg(-1)). Histological brain measurements were also carried out in a subset of the same rats. Neither LI nor PPI was altered using methods that had proven sensitive in previous pharmacological studies. Treated rats' locomotion was impaired, but amphetamine did not elicit a differential enhancement. A thinner Amon's horn layer was observed in their hippocampus. This indicates that standard models of schizophrenia did not fit to the behavioral abnormalities found by others and confirmed in this study. They were not due to the abnormal maternal care to pups elicited by the treatment. PMID- 15351507 TI - Identification of pleiotrophin in conditioned medium secreted from neural stem cells by SELDI-TOF and SELDI-tandem mass spectrometry. AB - Neural stem cells (NSCs) are multipotential progenitor cells that have self renewal activity. Since the fates of the NSCs in situ depend on their niche containing growth factors and cytokines, we performed surface enhanced laser desorption/ionization time-of flight mass spectrometry (SELDI-TOF-MS) to screen for differentially secreted proteins in conditioned medium of neural stem cells and compared with that of NIH3T3 cells. A 15.3-kDa protein detected only in the conditioned medium of neural stem cells was determined as pleiotrophin (PTN) by SELDI-TOF-MS and ProteinChip-tandem MS systems. Identification of pleiotrophin was further confirmed by one-dimensional SDS gel electrophoresis and Edman degradation analysis. The mRNA transcripts of PTN and its receptors [receptor protein tyrosine phosphatase (RPTP) beta/zeta, N-syndecan and anaplastic lymphoma kinase (ALK)] were detected in neurosphere, suggesting that pleiotrophin signaling systems are present in the neural stem cells and are involved in the modulation of fate of neural stem cells. PMID- 15351508 TI - Binge ethanol exposure delays development of GABAergic miniature postsynaptic currents in septal neurons. AB - Whole cell GABA(A)R currents of septal neurons isolated from rat pups increase rapidly during the first weeks of life when inhibitory synapses are forming. Early postnatal binge ethanol intubation on days 4-9 delays this maturational up regulation in septal neurons isolated several days later suggesting inhibitory synapse formation could be disrupted [S.-H. Hsiao, J.L. Acevedo, D.W. DuBois, K.R. Smith, J.R. West, G.D. Frye, Early postnatal ethanol intubation blunts GABA(A) receptor upregulation and modifies 3alpha-hydroxy-5alpha-pregnan-20-one sensitivity in rat MS/DB neurons, Brain Res. Dev. Brain Res. 130 (2001) 25-40]. Surprisingly, whole cell GABA(A)R function does not increase rapidly when septal neurons are grown for the same period in vitro and is not blunted by comparable ethanol exposure of the cultures [S.-H. Hsiao, D.W. DuBois, R.C. Miranda, G.D. Frye, Critically timed ethanol exposure reduces GABA(A)R function on septal neurons developing in vivo but not in vitro, Brain Res Dev. Brain Res. 1008 (2004) 69-80]. Because GABAergic miniature postsynaptic currents (mPSCs) show parallel patterns of maturation whether cortical neurons are growing in vivo or in vitro [D.D. Dunning, C.L. Hoover, I. Soltesz, M.A. Smith, D.K. ODowd, GABA(A) receptor-mediated miniature postsynaptic currents and alpha-subunit expression in developing cortical neurons, J. Neurophysiol. 82 (1999) 3286-3297], we examined the impact of binge ethanol exposure on synaptic receptors activated by these currents in septal cultures. Binge ethanol treatment of embryonic septal neurons over 6-11 days in vitro (DIV) slightly reduced GABA(A)R-mediated mPSC amplitude and frequency, but also substantially slowed decay kinetics when mPSCs were recorded later on DIV 13-18. Decreased frequency and slowed mPSC decay kinetics after ethanol were consistent with parameters measured in immature neurons. Untreated septal neurons exhibited decreased mPSC amplitude and frequency with acute 30-100 mM ethanol, without changing decay kinetics suggesting a direct inhibition of postsynaptic receptors. Sustained inhibition of GABA(A)Rs with 100 microM picrotoxin on DIV 6-11 decreased mPSC amplitude and frequency and slowed decay kinetics similar to binge ethanol exposure. These results suggest that binge ethanol exposure delays mPSC maturation by interfering with trophic postsynaptic GABA(A)R signaling during the early development of septal neurons. PMID- 15351509 TI - Predominant expression of GluR2 among the AMPA receptor subunits in neuronal progenitor cells of the rat hippocampus. AB - Cells derived from the hippocampus of embryonic day 18 (E18) rats were cultured in B27-supplemented Neurobasal medium without serum. We found the presence of numerous small cells with round or elliptical somata and fine processes in this primary culture. These cells were first detectable on culture day 8 and gradually increased in number that reached the maximum on approximately day 14. They incorporated bromodeoxyuridine (BrdU) and expressed nestin, a marker of stem cells and progenitor cells. Furthermore, nearly a half of these cells also expressed neuron-specific beta tubulin. On the other hand, they did not express O4 and glial fibrillary acid protein (GFAP), markers of oligodendrocytes and astrocytes, respectively. Thus, these small cells are most likely to be neuronal progenitor cells. The whole-cell patch clamp studies revealed that these cells expressed voltage-gated Na+, Ca2+ and K+ channels. With regard to ligand-gated channels, these cells were sensitive to alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA), but not to N-methyl-D-aspartate (NMDA). The current voltage relationship of the AMPA-induced current was slightly outwardly rectifying, suggesting that the AMPA receptors contained the GluR2 subunit in their oligomeric assemblies. The single-cell reverse transcription (RT)-PCR analysis revealed that GluR2 is predominant over the other AMPA receptor subunits in these cells. Furthermore, GluR2 was expressed mainly in the flip form. PMID- 15351510 TI - Clonidine increases caspase-3 mRNA level and DNA fragmentation in the developing rat brainstem. AB - The densities of alpha2-adrenergic receptors, labeled by 3H-clonidine or 3H RX821002, reach a peak in the rat brainstem during the first week of its life. This enables the agonist of alpha2-adrenergic receptor clonidine, which is used as a component of anaesthetic solution in infants and children, to have specific effects in this structure of the developing brain. Clonidine was injected into the fetal brain (5 microg in 5 microl of saline) or subcutaneously to the pups (1, 10 microg in 50 microl of saline) 3 days before investigation. Clonidine increased the level of apoptotic enzyme caspase-3 mRNA expression, as measured by RT-PCR and enhanced the DNA fragmentation, as determined by gel electrophoresis, in the brainstem of the 21-day-old fetuses and 8-day-old rats. In the cortex of 8 day-old rat, the alpha2-adrenergic receptors are at a much lower level than the brainstem. Clonidine treatment had no evident effects on caspase-3 mRNA level and DNA fragmentation in the cortex of an 8-day-old rat. The data suggest that clonidine facilitates cell death in the developing brainstem. This drug effect provides a potential mechanism whereby clonidine during early life could induce long-lasting alterations in brain neurochemistry, autonomic functions and behavior. PMID- 15351511 TI - Transiently overexpressed alpha2-adrenoceptors and their control of DNA synthesis in the developing brain. AB - During brain development, neurotransmitters act as trophic factors controlling the patterns of cell replication and differentiation. Alpha2-adrenoceptors (alpha2ARs) are transiently overexpressed in zones with high mitotic activity and we evaluated whether these receptors are linked to DNA synthesis in the perinatal rat brain. Acute administration of clonidine (2 mg/kg), an alpha2AR agonist, elicited dramatic decreases in DNA synthesis in the forebrain, brainstem, and cerebellum whether given on gestational day (GD) 21, or on postnatal days (PN) 1 or 8. However, alpha2AR blockade elicited by yohimbine (2.5 mg/kg) also resulted in decreased DNA synthesis on GD21 and PN8, albeit to a smaller extent than with clonidine. Yohimbine was able to blunt the effects of clonidine, verifying that both drugs are acting through the same receptor population. Because betaARs are also known to regulate DNA synthesis, we used propranolol (10 mg/kg) blockade of betaARs to evaluate whether the alpha2AR effects were mediated by presynaptic autoreceptors that regulate the release of norepinephrine and consequent betaAR responses; the effects of yohimbine were still discernible in the presence of propranolol. Accordingly, transiently overexpressed alpha2ARs in the developing brain participate in the control of DNA synthesis in a biphasic manner, with promotional actions at low, endogenous levels of stimulation, but inhibitory effects when stimulation is high. Effects on alpha2ARs are likely to contribute to long-term consequences of adrenergic agents used in obstetrics or neurotoxicants that affect adrenergic activity. PMID- 15351512 TI - The influx of neutral amino acids into the porcine brain during development: a positron emission tomography study. AB - Pigs of three different age groups (newborns, 1 week old, 6 weeks old) were used to study the transport of the large neutral amino acids 6-[18F]fluoro-L-DOPA ([18F]FDOPA) and 3-O-methyl-6-[18F]fluoro-L-DOPA ([18F]OMFD) across the blood brain barrier (BBB) with positron emission tomography (PET). Compartmental modeling of PET data was used to calculate the blood-brain clearance (K1) and the rate constant for the brain-blood transfer (k2) of [18F]FDOPA and [18F]OMFD after i.v. injection. A 40-70% decrease of K1(OMFD), K1(FDOPA) and k2(OMFD) from newborns to juvenile pigs was found whereas k2(FDOPA) did not change. Generally, K1(OMFD) and k2(OMFD) are lower than K1(FDOPA) and k2(FDOPA) in all regions and age groups. The changes cannot be explained by differences in brain perfusion because the measured regional cerebral blood flow did not show major changes during the first 6 weeks after birth. In addition, alterations in plasma amino acids cannot account for the described transport changes. In newborn and juvenile pigs, HPLC measurements were performed. Despite significant changes of single amino acids (decrease: Met, Val, Leu; increase: Tyr), the sum of large neutral amino acids transported by LAT1 remained unchanged. Furthermore, treatment with a selective inhibitor of the LAT1 transporter (BCH) reduced the blood-brain transport of [18F]FDOPA and [18F]OMFD by 35% and 32%, respectively. Additional in vitro studies using human LAT1 reveal a much lower affinity of FDOPA compared to OMFD or L-DOPA. The data indicate that the transport system(s) for neutral amino acids underlie(s) developmental changes after birth causing a decrease of the blood-brain barrier permeability for those amino acids during brain development. It is suggested that there is no tight coupling between brain amino acid supply and the demands of protein synthesis in the brain tissue. PMID- 15351513 TI - Expression of Mcl-1 in cerebellar granule neurons is regulated by IGF-I in a developmentally specific fashion. AB - Transgenic (Tg) mice that overexpress IGF-I during postnatal brain development exhibit remarkable cerebellar overgrowth characterized by significant increases in granule cell number that is predominantly due to IGF-I anti-apoptotic actions. Using these mice as a model to define the gene expression profile underlying the pro-survival actions of IGF-I, we screened 243 apoptosis-related genes by cDNA arrays and found that Mcl-1 was down-regulated in cerebella of IGF-I Tg mice. Contrary to the results obtained by cDNA array, Northern blot analyses showed that the Mcl-1 mRNA abundance in the cerebella of IGF-I Tg mice at postnatal day 14 (P14) was five times more than that of wild-type (Wt) controls. The increase in Mcl-1 mRNA expression in IGF-I Tg mice was detected as early as P8, peaked at P14, and remained detectable at P20. Both IGF-I Tg and Wt mice showed a similar expression pattern of Mcl-1 mRNA which coincided with the post-mitotic migration and the post-migratory maturation of granule cells. We measured the relative abundance of Mcl-1 protein in the cerebellum by immunoblots and found that anti apoptotic Mcl-1L was the predominant form, while pro-apoptotic Mcl-1S was minimally detectable. Cerebellar Mcl-1L was 2.6-fold more abundant in IGF-I Tg mice compared with that in their Wt littermates. Using laser capture microdissection followed by RT-PCR, we determined that Mcl-1 mRNA was expressed in granule cells, but not in Purkinje cells. In summary, these findings show that the anti-apoptotic Mcl-1 isoform is expressed in cerebellar granule neurons, which undergo apoptosis during postnatal cerebellar cortical lamination, and Mcl 1 expression is up-regulated by IGF-I overexpression in a developmentally specific manner. These data suggest that anti-apoptotic Mcl-1 may mediate IGF-I pro-survival actions on granule neurons during the development of cerebellar cortex. They also point out pitfalls of cDNA array analyses. PMID- 15351514 TI - Identification of a mouse orthologue of the G-protein-coupled receptor SALPR and its expression in adult mouse brain and during development. AB - The mouse orthologue of somatostatin and angiotensin-like peptide receptor (SALPR) was amplified from cDNA of the hippocampal cell line HT22. It coded for a protein of 472 amino acids showing 84% sequence identity with human SALPR and 43% with human G-protein-coupled receptor 100 (GPR100). A distinct pattern of expression in brain, spinal cord, and dorsal root ganglia during development and in the mature brain hint at important functions of SALPR for differentiation and maintenance of the nervous system. PMID- 15351515 TI - Integrin and the Reelin-Dab1 pathway: a sticky affair? AB - It has been repeatedly suggested that integrin is essential for neuronal migration. A new study proposes a link between Disabled-1 (Dab1) phosphorylation and alpha3 integrin signalling that is thought to drive the timely detachment of migrating neurons from the guiding radial glia fibers during early corticogenesis. This proposal however is hardly compatible with time-lapse anatomical investigations and genetic studies on integrin or with recent works which indicate direct reelin signalling to radial fibers. PMID- 15351516 TI - Improved immunocytochemical identification of neural, endothelial, and inflammatory cell types in paraffin-embedded injured adult rat spinal cord. AB - Methods that facilitate the accurate counting of specific neural cell types would be of substantial value in evaluating the efficacy of treatments applied to spinal cord injury. This report describes reliable procedures for identification of neurons, oligodendrocytes, astrocytes, endothelial cells and inflammatory cells (neutrophils and activated macrophage/microglial cells) in paraformaldehyde fixed, paraffin-embedded injured adult rat spinal cord. Antigen retrieval techniques (enzymatic and thermal) were used to improve antibody access to masked epitopes. To decrease background immunofluorescence and autofluorescence of hemoglobin, the tissue sections were pretreated with 0.1% sodium borohydride in PBS (30min), followed by 1-5min incubation in 0.5% Sudan black in 70% ethanol. Commercially available techniques to amplify the primary signal such as tyramide signal amplification (TSA) and avidin/biotin/peroxidase/DAB/nickel/cobalt amplification (ABP/DABA) were also tested. Hoechst 33342 nuclear staining was used to indicate cell location, number, and integrity, thereby avoiding misidentification of cells. The best antibodies were: anti-NeuN antibody for neurons, anti-S100 for astrocytes, and anti-S100 and APC-7 antibodies in combination for oligodendrocytes, anti-laminin (LN) for endothelial cells, and ED1 antibody for activated macrophages and microglia. Amplification of the primary signal with TSA or ABP/DABA was also found to be beneficial. PMID- 15351517 TI - 3D-catFISH: a system for automated quantitative three-dimensional compartmental analysis of temporal gene transcription activity imaged by fluorescence in situ hybridization. AB - Fluorescence in situ hybridization (FISH) of neural activity-regulated, immediate early gene (IEG) expression provides a method of functional brain imaging with cellular resolution. This enables the identification, in one brain, of which specific principal neurons were active during each of two distinct behavioral epochs. The unprecedented potential of this differential method for large-scale analysis of functional neural circuits is limited, however, by the time-intensive nature of manual image analysis. A comprehensive software tool for processing three-dimensional, multi-spectral confocal image stacks is described which supports the automation of this analysis. Nuclei counterstained with conventional DNA dyes and FISH signals indicating the sub-cellular distribution of specific, IEG RNA species are imaged using different spectral channels. The DNA channel data are segmented into individual nuclei by a three-dimensional multi-step algorithm that corrects for depth-dependent attenuation, non-isotropic voxels, and imaging noise. Intra-nuclear and cytoplasmic FISH signals are associated spatially with the nuclear segmentation results to generate a detailed tabular/database and graphic representation. Here we present a comprehensive validation of data generated by the automated software against manual quantification by human experts on hippocampal and parietal cortical regions (96.5% concordance with multi-expert consensus). The high degree of reliability and accuracy suggests that the software will generalize well to multiple brain areas and eventually to large-scale brain analysis. PMID- 15351518 TI - Sindbis viral-mediated expression of eGFP-dopamine D1 receptors in situ with real time two-photon microscopic detection. AB - Dopamine D1 receptors (D1DRs) mediate a major component of dopaminergic neurotransmission, and alterations in their synaptic and subcellular distribution may underlie a variety of neurological diseases. In order to monitor D1DR localization in real time, we subcloned a sindbis virus containing an enhanced GFP coding region inserted at the C-terminal region of a dopamine D1 receptor (eGFP-D1DR). Two-photon excitation of expressed eGFP-D1DRs was monitored in a variety of viable neural preparations. Infection of primary cultured rat ventral striatal neurons, verified for neuronal phenotype using patch clamp electrophysiology, was induced by the simple addition of the virus to media. Parasagittal slice cultures, including the ventral tegmental area (VTA) and nucleus accumbens (NAc), were infected by manual injection below the glia surface. NAc-containing parasagittal slices prepared from mice in which the virus was administered via stereotaxic injection in vivo also displayed robust eGFP D1DR expression. Expression of functional D1DRs following infection in baby hamster kidney (BHK) cells was monitored by DA-stimulated cAMP production that was also blocked by a selective D1 antagonist. Taken together, these findings provide the first demonstration of the functional expression and real-time imaging of eGFP-D1DRs, and indicate that sindbis virus is an effective method for D1 receptor expression in a variety of native neuronal preparations. PMID- 15351519 TI - A reliable technique for the induction of locomotor-like activity in the in vitro neonatal rat spinal cord using brainstem electrical stimulation. AB - The use of brainstem electrical stimulation (BES) to evoke locomotion in in vivo preparations, such as the decerebrate cat, is well established. In contrast, despite the popularity of in vitro rodent spinal cord models, BES has not been adapted for routine induction of locomotion in these preparations. We describe a simple and reliable method of inducing locomotor-like activity in the in vitro neonatal rat spinal cord using BES. Relatively large amplitude (0.5-10mA), long duration (4-20ms) and low frequency (0.8-2.0Hz) pulses were delivered through a metal-in-glass electrode placed in contact with the ventral surface of the brainstem, without the need for precise targeting of specific sites. During continuous BES, locomotor-like activity (0.15-0.63Hz) persisted for over 45min. Episodes of locomotor-like activity could be recruited repeatedly for hours, using short periods (60s) of BES alternating with brief rest periods. Additional observations confirmed that the rhythmogenic effect of BES is mediated by excitation of spinal projections at the brainstem level, rather than spinal cord activation due to electrotonic spread of stimulus from the brainstem electrode. Endogenous activation of locomotor networks using BES offers important advantages over bath-applied application of neurochemicals to induce stepping in the in vitro rat model. PMID- 15351520 TI - A new strategy for analysis of phenotype marker antigens in hollow neurospheres. AB - Changes in expression of phenotypic markers characterizing the cells comprising intact neurospheres are difficult to determine easily and accurately. The problem is compounded by the diversity of cell populations and phenotypes involved, and consequent non-uniform protein expression across the neurosphere wall, or around the circumference. Therefore, interpreting the effects of phenotype modifying conditions has been a complex and demanding task. Here, we report a novel direct method for measuring protein expression in immunofluoerescently labeled intact neurospheres by densitometric image analysis of optical cross-sections, obtained by scanning laser confocal microscopy. To demonstrate our methodology, hollow human neurospheres were exposed to basic fibroblast growth factor (FGF2), which reportedly induces neuronal commitment in monolayer cultures of neuroprogenitor cells derived from neurospheres. We determined that this treatment downregulated nestin and vimentin, protein markers accepted to indicate an immature, uncommitted phenotype. Neuron specific enolase was only marginally affected. Our strategy allows quantitation of changes in expression of marker proteins that is comparable to Western blot analysis. In addition to discriminating heterogeneity in protein expression, suitable optics may allow the resolution down to single cell level. We propose that this novel strategy, with or without confocal microscopy, may be applied to other biological systems. Analysis of protein expression by the cells comprising tubular or cylindrical cellular structures, or approximately spherical cell aggregates, can be performed efficiently using a small sample size. PMID- 15351521 TI - Real-time visualization of neural synchrony for identifying coordinated cell assemblies. AB - We introduce a synchrony map that translates the fine temporal organization of multi-unit responses in the visual cortex into an easily interpreted spatial display. We test the synchrony map on microelectrode array recordings in Area 17 of anesthetized and paralyzed cats. We first examine the synchrony map using averaged data and probability calculations to demonstrate orientation-dependent changes in synchrony. We then demonstrate how the synchrony map can be implemented for real-time visualization of synchrony among neural assemblies. PMID- 15351522 TI - Beta-escin diminishes voltage-gated calcium current rundown in perforated patch clamp recordings from rat primary afferent neurons. AB - Perforated patch recordings of neuronal calcium currents (I(Ca)) with amphotericin B or nystatin reduce dialysis of intracellular constituents and current rundown, but can be difficult and frequently unsuccessful. We investigated the saponin beta-escin as a putative ionophore for perforated patch I(Ca) recordings in acutely dissociated, rat dorsal root ganglion neurons. I(Ca) was recorded in time-course studies after including either beta-escin (50 microM), or amphotericin B (240 microg/ml) as perforating ionophores in the internal pipette solution, in comparison to standard ruptured-patch technique, using suction. Perforated patches were allowed to take place spontaneously. The percentage loss of I(Ca) per min (within the first 20 min) was significantly less after beta-escin (0.0518%) (n = 18), versus either amphotericin (1.82%) (n = 12) or standard patch (4.52%) (n = 7), (P < 0.001). The slope of the rundown after linear fit was also less after beta-escin (P < 0.001). Minimal "steady-state" access resistance (R(a)) of 6.6 +/- 1.6 MOmega was achieved within 7.1 +/- 9.3 min following perforation with beta-escin, 7.9 +/- 3.5 MOmega within 44 =/- 14 min after amphotericin B, and 6.8 +/- 1.9 MOmega with standard patch (P < 0.05 for R(a), and P < 0.01 for permeabilization time, respectively). Success rates were 59% with beta-escin versus 27% with amphotericin. Leak >10% of peak I(Ca) was present in 25% of cells after beta-escin versus 20% after amphotericin, and 12% after standard technique. Perforated patches using beta-escin were stable for 15-60 min. We conclude that beta-escin may be used as an alternative ionophore for perforated patch-clamp studies in neurons, and results in minimal rundown that can facilitate long-term recordings of I(Ca). Limited rundown may be due to better preservation of cytosolic ATP content. PMID- 15351523 TI - A novel method of thermal activation and temperature measurement in the microscopic region around single living cells. AB - We present a simple approach to bring fast and reversible temperature steps of a wide range of amplitudes from the temperature of the experimental chamber up to the boiling point of water in a desired position, with rise and fall times of around 10 ms in a microvolume of microm in size, such as in a single cell. For this purpose, we applied a technique for illuminating a metal aggregate (1-2 microm in diameter) placed at the tip of a glass micropipette with a focused infrared (1064 nm) laser beam under an optical microscope. Stable temperature gradients were created around the metal aggregate using an appropriate neutral density filter set for the laser output. To monitor the local temperature, we devised a new microthermometer composed of the tip of a micropipette filled with thermosensitive fluorescent dye Europium-TTA possessing steep temperature dependent phosphorescence upon 365 nm excitation. The microm size of the tip of this pipette was able to measure the local temperature with 0.1 degrees C precision and microm spatial resolution. This new approach is compatible with standard electrophysiological and imaging techniques. PMID- 15351524 TI - Method for identifying neuronal cells suffering zinc toxicity by use of a novel fluorescent sensor. AB - During excitotoxic brain damage, injured neurons accumulate an anomalous, pathological burden of weakly bound, rapidly exchangeable Zn(2+) that diffusely fills the soma, nucleus and proximal dendrites. Mounting evidence indicates that this Zn(2+) is a major contributing factor in the subsequent demise of the damaged neurons. Thus, identifying, imaging, and characterizing zinc-filled cells have become essential steps in understanding excitotoxicity. Here we demonstrate that a new fluorescent stain for zinc can rather selectively and quite vividly label zinc-filled neurons in frozen histologic sections. The method is more sensitive and selective than the existing stain TSQ, and simpler than the Timm Danscher silver staining techniques. A previously unobserved population of apparently injured cells in the dentate gyrus has been discovered with the new reagent. Whereas cells viewed in situ in normal, healthy tissue virtually never display any perikaryal staining by histochemical methods for zinc, injured cells stain intensely for zinc in culture, acute slice preparations and in tissue harvested in vivo. Thus, the presence of rapidly-exchangeable, "stainable" perikaryal zinc may be taken as an indicator of cell injury. PMID- 15351525 TI - Analysis of functional MRI data based on an estimation of the hemodynamic response in the human brain. AB - Functional magnetic resonance imaging (fMRI) data is usually analyzed using hemodynamic response data formulated with the aid of a gamma function. An fMRI response to stimuli can be mathematically modeled by convolution of the hemodynamic response and the presented stimulus. This approach is based on a linear system analysis. However, it is known that most biological systems are nonlinear and the nature of the hemodynamic response depends on both the subjects and brain regions under study. In this work, we estimated the hemodynamic responses of the fusiform face area (FFA) and the primary visual area (V1) using 1st- and 2nd-order Volterra kernels. The estimated hemodynamic responses were used to analyze fMRI data obtained from the corresponding regions. The results of the analysis of fMRI data using the estimated hemodynamic responses show a more significant activation than is obtained by conventional analysis using the hemodynamic response modeled by gamma function. PMID- 15351526 TI - A laser micromachined probe for recording multiple field potentials in the thalamus. AB - Multichannel recording provides integral information about electrical brain activities at one instant in time. In this study, multielectrode probes were fabricated to record the thalamic field potentials (FPs) responding to the electrical stimulation of nerve at the rat tail. At first, the number of sweeps used to form the evoked FP average and the spatial sampling density were determined by using cross-correlation functions, which were then statistically analyzed. The difference was significant at P < 0.05, if the number of sweeps for averaging was more than 50 and the spatial interval between two consecutive recording sites was less than 50 microm in the anteroposterior, mediolateral and ventrodorsal directions. The responsive area was distributed vertically in the thalamus (ventral posterior lateral (VPL) nucleus); therefore, the recording sites were arranged in one linear array. Sixteen recording sites, which were 50 microm apart from each other, were distributed in the ventrodorsal direction. A 16-channel silicon probe was fabricated by using a standard photolithography process and laser micromachining techniques. The probe provides capabilities to record multiple thalamic evoked FPs and multiunit activities simultaneously. PMID- 15351527 TI - Multivariate tests for the evaluation of high-dimensional EEG data. AB - In this paper several multivariate tests are presented, in particular permutation tests, which can be used in multiple endpoint problems as for example in comparisons of high-dimensional vectors of EEG data. We have investigated the power of these tests using artificial data in simulations and real EEG data. It is obvious that no one multivariate test is uniformly most powerful. The power of the different methods depends in different ways on the correlation between the endpoints, on the number of endpoints for which differences exist and on other factors. Based on our findings, we have derived rules of thumb regarding under which configurations a particular test should be used. In order to demonstrate the properties of different multivariate tests we applied them to EEG coherence data. As an example for the paired samples case, we compared the 171-dimensional coherence vectors observed for the alpha1-band while processing either concrete or abstract nouns and obtained significant global differences for some sections of time. As an example for the unpaired samples case, we compared the coherence vectors observed for language students and non-language students who processed an English text and found a significant global difference. PMID- 15351528 TI - Reasons given by relatives of colorectal cancer patients for not undergoing screening. AB - BACKGROUND: Relatives of colorectal cancer (CRC) patients are at increased risk of developing CRC themselves. Although this increased-risk group could benefit from screening, many do not undergo screening. Little is known about the reasons why this group chooses not to be screened, and understanding these reasons has implications for the development of tailored interventions. METHODS: Telephone interviews were conducted with 132 nonscreening relatives of CRC patients. Variables collected included reasons for not screening, intent to be screened in the future, potential prompts to future screening, and perceived benefits and barriers to screening as well as perceived CRC risk. RESULTS: We were able to categorize the majority of nonscreeners into four discrete types: a group who cite feeling healthy as their reason for not screening (n = 40), a group whose physicians have not endorsed screening (n = 18), a younger group who were told to wait to be screened (n = 21), and a group who anticipate discomfort and pain with screening (n = 20). CONCLUSIONS: It is possible to identify distinct groups of nonscreeners according to their particular barriers and reasons for not screening. Tailored interventions can be designed to address the specific issues in each of these groups. PMID- 15351529 TI - Cancer screening among older women in a culturally insular community. AB - BACKGROUND: "Culturally insular communities" are defined by a religious or ethnic orientation that effectively places them outside mainstream sources of information on health promotion. We investigated whether older women in one such community were less likely to be screened for breast, cervical, and colorectal cancer than other women in the same geographic area. METHODS: A random sample of older orthodox Jewish women in Borough Park, Brooklyn, was compared to white women of the same age in the New York metropolitan area who were surveyed in the Behavioral Risk Factors Surveillance System (BRFSS). RESULTS: Borough Park women aged 50-69 were less likely to report they had been screened for colorectal cancer, and women over age 70 were less likely to report screening for all three cancer prevention modalities. Among Borough Park women, functional and cognitive deficits, elicited by self-report, also reduced the likelihood of screening. CONCLUSIONS: Cultural and religious prescriptions are important to consider in health promotion. These factors should be taken into account in designing strategies to promote cancer screening. PMID- 15351530 TI - Greek employee awareness of carcinogenic exposure. AB - BACKGROUND: Occupational risk factors contribute significantly to the development of lung cancer; however, little is known about the extent to which employees are informed of occupational exposure to carcinogenic substances. METHODS: Through a case-control study, we estimated the level of awareness among Greek employees potentially exposed to known carcinogenic substances within various occupational settings. RESULTS: Only 6.6% of men (n = 482) employed in occupations with potential exposure to carcinogenic substances were aware of such occupational exposures. Age, education, and residence were significantly associated with awareness. Employees having at least a secondary level of education were 3.5 times more aware than those having at most 6 years of educational training. CONCLUSIONS: Assessing awareness among workers potentially exposed to occupational risk factors and promoting occupational health education are important steps for increasing health and safety at the workplace. PMID- 15351531 TI - Cigarette smoking among rural-to-urban migrants in Beijing, China. AB - BACKGROUND: Tobacco use remains a substantial public health challenge in China. The rapidly growing rural-to-urban migrant population currently numbering approximately 121 million individuals may be especially vulnerable to tobacco use. METHODS: Cigarette smoking levels and associated factors were examined among 2,201 subjects using cross-sectional data from the Migrant Health Behavior Survey (MHBS), conducted in 2002, Beijing, China. RESULTS: Respondents (39.3% females) were 18-30 years old with a mean age of 22.7 years; 77.3% were single and 35.6% had at least attended high school for education. Migrants reported an average of 4 years of migratory experience; 28.8% had worked in at least three cities and 36.8 had held at least four jobs. Males (51.7%) and females (10.9%) reported 30 day cigarette use. Smoking levels were positively associated with age, education, income, migration years, number of cities in which the migrants had worked, living in rental properties, and number of jobs held. Smoking levels were negatively associated with job and life satisfaction and living with relatives in Beijing. Smoking levels were not associated with marital status. CONCLUSIONS: Female migrants had disproportionately high rate of cigarette smoking. Several conditions and experiences common to the migratory lifestyle were associated with increased levels of smoking. Tobacco control measures are needed to protect migrant populations in China. PMID- 15351532 TI - Attitudes and behaviors related to weight control in two diverse populations. AB - BACKGROUND: Researchers have found that African-American women have concerns about their weight, but are less successful in their weight loss efforts compared to Caucasian women. Reasons for this disparity have not been explored in great detail. Identifying specific attitudes and behaviors that contribute to obesity in these subpopulations will help inform the design of interventions. METHODS: This report compares specific attitudes and behaviors related to diet and exercise in two groups who are trying to lose weight: African-American women in Indianapolis (n = 80) and Caucasian women from the rural Midwest (n = 45). Data are from surveys that shared many common measures, but were otherwise not related to one another. The choice of measures was guided by a conceptual framework of self-management processes. RESULTS: Compared to women in the urban study, women in the rural study report greater social support for behavior change, greater self-efficacy, and outcome expectations, and greater use of every strategy measured. CONCLUSIONS: While there is room for improvement in both groups, these results contribute to our understanding of subgroup differences and similarities in the complex behavioral issues surrounding successful weight management. Larger studies with more representative samples are needed to confirm these findings and inform future interventions. PMID- 15351533 TI - Effects of hiking at altitude on body composition and insulin sensitivity in recovering drug addicts. AB - In the current study individuals with a history of drug abuse (users of heroin, cocaine, or amphetamine) displayed a 13-100% increase in body weight (self reported) and exhibited a trend toward insulin resistance. Therefore, we investigated the effects of long-term altitude hiking on insulin sensitivity in this special population. Nine males recovering from drug addiction (ex-addicts) (age 28.7 +/- 1.3 years) and 17 control subjects (age 29 +/- 1.1 years) voluntarily participated in a 25-day hiking activity (altitude 2200-3800 M). On the 25th day of hiking, oral glucose tolerance test (OGTT), insulin response, lean body mass, fat mass, and waist-to-hip ratio (WHR) were measured in all subjects. After the altitude expedition, insulin levels during the OGTT in ex addicts were similar to controls, suggesting that insulin sensitivity in this special population was normalized by long-term altitude activity. Along with improvements in insulin sensitivity, a significant reduction in WHR, but small increase in lean body mass, was observed. Twenty-five days of altitude activity significantly reverses hyperinsulinemia in the ex-addicts and this improvement appears to be partially associated with the reduction in central fatness. PMID- 15351534 TI - Periconceptional folate intake by supplement and food reduces the risk of nonsyndromic cleft lip with or without cleft palate. AB - BACKGROUND: Inadequate maternal vitamin intake during pregnancy has been suggested as a risk factor for cleft lip with or without cleft palate (CLP). The independent role of folate has not been clarified. METHODS: To investigate the association between maternal folate intake by supplement and food and the risk of CLP offspring, a case-control study was conducted in the Netherlands (1998-2000) among 174 mothers of a child with nonsyndromic CLP and 203 mothers of a child without congenital malformations. RESULTS: Daily use of a folic acid supplement by mothers starting from 4 weeks before until 8 weeks after conception gave a 47% CLP risk reduction compared to mothers who did not use these supplements [odds ratio (OR): 0.53, 95% confidence interval (CI): 0.33, 0.85]. Ninety-three percent of the users took a supplement containing folic acid only. Dietary folate intake reduced CLP risk independently in a dose-response manner. The largest risk reductions were found on those mothers who had a diet of more than 200 microg folate per day in combination with a folic acid supplement (OR: 0.26, 95% CI: 0.09, 0.72). CONCLUSIONS: We demonstrated that periconceptional maternal folic acid supplement use was beneficial to reduce the risk for CLP. An additional effect of food folate was shown. PMID- 15351535 TI - Identification of factors promoting abstinence from smoking in a cohort of elementary schoolchildren. AB - BACKGROUND: Despite the progress made in smoking prevention among youth, few scientific reports have identified the determinants underlying the abstinence from cigarettes, and even fewer have made an assessment of their evolution over time. It has been proposed that this could be done by modeling after theoretical frameworks. METHODS: A self-reported questionnaire developed according to Ajzen's Theory of Planned Behavior was repeatedly administered to a cohort of 373 grade 5 students (191 girls and 182 boys) during their transition from elementary to secondary school. Four measures spanning a 26-month period were thus obtained and analyzed according to the GEE method. RESULTS: After 26 months, 76.14% of the students in the sample remained abstinent from cigarettes. The perceived smoking behaviors of friends and of a brother, intention over time, perceived self efficacy, parental supervision, and socioeconomic status predicted the maintenance of abstinence from smoking. The final predictive model had a sensitivity of 79.8% and a specificity of 75.6%, with an average rate of accurate classification of 79.1%. CONCLUSIONS: The predictive model yielded by this study represents an important step toward developing tools to help 10-12 years olds maintaining their abstinent behavior. It emphasizes positive protective factors and guides health promoters in the choice of their interventions' contents and targets. PMID- 15351536 TI - County characteristics and racial and ethnic disparities in the use of preventive services. AB - BACKGROUND: Studies examining predictors of preventive service utilization generally focus on individual characteristics and ignore the role of contextual variables. To help address this gap in the literature, the present study investigates whether county-level characteristics, such as racial and ethnic composition, are associated with the use of preventive services. METHODS: Data from the Medical Expenditure Panel Survey and the Area Resource Files (1996-1998) are used to identify the individual- and county-level predictors of five types of preventive services (n = 49,063). RESULTS: County racial or ethnic composition is associated with the utilization of certain preventive services, net of individual level characteristics. Specifically, individuals in high percent Hispanic counties are more likely to report cholesterol screenings, while those in counties with more blacks are more likely to have regular mammograms. Moreover, county racial or ethnic composition modifies the relationship between individual race or ethnicity and preventive use. In particular, Hispanic individuals who reside in high percent black counties report higher levels of utilization for most preventive services compared to Hispanics living in other counties. CONCLUSIONS: Physical and social environments are key determinants of health behaviors and outcomes. Future studies should take into account the racial or ethnic composition of an area and how this interacts with individual race or ethnicity when investigating predictors of preventive care use. PMID- 15351537 TI - Risks of anemia in infants according to dietary iron density and weight gain rate. AB - BACKGROUND: The objective of the study was to identify risk factors of infant anemia and to assess a new indirect method to identify the risks of anemia using a probability curve based on dietary iron density and weight gain rate. METHODS: A cross-sectional study was conducted on 110 term infants in a public health unit in Goiania, Brazil. A socioeconomic, demographic, anthropometric survey and dietary assessment were performed. Hemoglobin, red blood cell distribution width (RDW), ferritin, and C-reactive protein were determined. RESULTS: The risk factors identified by final adjusted logistic regression were male infants (OR = 2.38), infants whose mothers were housewives (OR = 5.01), and households with more than three members (OR = 3.08). Another risk factor was the introduction of meat after 6 months (OR = 3.3). There was a positive dose-response effect between milk consumption and prevalence of anemia adjusted for sex and age (Effect = 0.120; 95% CI: 0.015, 0.225). Dietary iron density (from 6 to 9 months) and weight gain rate (from 9 to 12 months) significantly affected the probability of the infant having anemia. CONCLUSIONS: The probability curve for anemia obtained from dietary iron density and weight gain rate can be used to identify the risks of anemia, and together with the risk factors identified are important for anemia prevention programs. PMID- 15351538 TI - Changes in health-related behaviours and cardiovascular risk factors in young adults: associations with living with a partner. AB - OBJECTIVE: To examine changes in cardiovascular risk factors and health-related behaviours in young Australian adults at a stage of transition from the family environment. STUDY DESIGN: Repeated surveys between 9 and 25 years of age in a community-based group that included 569 eighteen-year-olds and 600 twenty-five year-olds. RESULTS: There were significant increases (P < 0.001 for all variables) in body mass index (BMI) [men 2.5 kg/m2 (2.0); women 1.7 kg/m2 (2.9)], waist girth [men 7.6 cm (6.5); women 4.3 cm (7.2)], BP (systolic/diastolic) [men 5(12)/7(8) mm Hg; women 3(10)/6(7) mm Hg] and in total cholesterol [men 15% increase; women 9%]. The proportion of sedentary behaviour increased from 19% to 39% in men (P < 0.001) and from 40% to 41% (P = 0.801) in women. Cohabitation was associated with significantly greater increases in BMI, waist circumference, and total cholesterol, associated with dietary change in women and decreased physical activity in men. In mothers, waist girth increased by 8.0 cm (0.1) compared with 3.5 cm (0.6) in women without children (P = 0.003), and physical fitness decreased [-0.5 W/kg (0.4) vs. 1.2 W/kg (0.2), respectively; P = 0.001]. CONCLUSION: Encouragement of a healthy lifestyle, particularly physical activity, should be a priority in this age group, particularly among newly cohabiting couples and in young mothers. PMID- 15351539 TI - Increased awareness, knowledge and utilization of preconceptional folic acid in Israel following a national campaign. AB - BACKGROUND: To decrease the risk of neural tube defects (NTDs), the Israeli Ministry of Health (MOH) issued guidelines in August 2000 recommending daily folic acid (FA) supplementation for women in their childbearing age, and concurrently launched a national FA campaign. Campaign effects were assessed by comparing the results of a survey done in 2002 with a baseline survey done in June 2000. METHODS: Both surveys were done within the network of the Public Health Services' Mother and Child Health Clinics (MCHC). Nursing staff conducted structured interviews of pregnant women and mothers of newborn infants. RESULTS: In the 2002 survey (n = 1661), awareness was 85%, correct knowledge was 77.7% and 30.5% utilized FA preconceptionally. Ratios of awareness, knowledge and utilization were highest among women with post-university education (93%, 84%, 46%), and awareness and utilization were significantly higher in the 25-29 year age bracket (90%, 35%). In the baseline 2000 survey (n = 1719), FA awareness had been 54.6%, knowledge of the benefits of FA was 17.6% and preconceptional utilization was reported by a mere 5.2%. CONCLUSIONS: A national periconceptional FA campaign in Israel resulted in significant increases in awareness and correct knowledge, and a sixfold increase in its intake. PMID- 15351540 TI - Sociodemographic differences in use of preventive services by women enrolled in Medicare+Choice plans. AB - BACKGROUND: We examined the effect of sociodemographic factors on the receipt of mammography, colorectal cancer screening, and influenza vaccinations by women enrolled in two Medicare+Choice health plans. METHODS: Administrative and survey data for 2,698 female health plan members was analyzed using multivariate logistic and ordinal logistic regression to assess the effects of enrollee characteristics on use of preventive services. RESULTS: Age, race and wealth were associated with the receipt of one or more preventive services. Older women were less likely to receive mammograms, wealthier women were more likely to receive mammograms and CRC screening, and Black women were more likely to receive CRC screening but less likely to receive influenza vaccinations. Wealthier women received a greater number of preventive services, other things equal, while older women received fewer preventive services. CONCLUSIONS: Race and wealth continue to be important factors in the receipt of preventive services by elderly women, though not always consistent with historical trends. Medicare+Choice plans should consider strategies to further reduce racial and wealth disparities in the use of preventive services. PMID- 15351541 TI - Relationship of diabetes-specific knowledge to self-management activities, ambulatory preventive care, and metabolic outcomes. AB - BACKGROUND: Educational interventions increase diabetes patients' knowledge and self-care activities, but their impact on the use of health services to prevent diabetes complications is unclear. We sought to determine the relationship of patients' diabetes-specific knowledge with self-management behaviors, use of ambulatory preventive care, and metabolic outcomes. METHODS: We surveyed 670 adults with diabetes from three managed care plans to assess diabetes knowledge (using an eight-item scale) and self-management activities. With chart review, we assessed five processes of care--retinal and foot examinations, low-density lipoprotein cholesterol (LDL-C) testing, hemoglobin A1c (HbA1c) testing, and urine microalbumin testing--and three metabolic outcomes--HbA1c < or = 9.5%, LDL C <130 mg/dL (3.36 mmol/L), and last blood pressure <140/90 mm Hg. RESULTS: In adjusted analyses, a one-point increase on the knowledge scale was associated with following a diabetes diet (OR 1.23, 95% CI 1.10-1.38), blood glucose self measurement (OR 1.29, 95% CI 1.13-1.48), and regular exercise (OR 1.15, 95% CI 1.03-1.28) but not with processes of care or metabolic outcomes. CONCLUSIONS: Knowledgeable patients were more likely to perform self-management activities but not to receive recommended ambulatory care or reach metabolic outcome goals. Providing patient education about diabetes care processes should be tested as a means of increasing ambulatory care to prevent diabetes complications. PMID- 15351542 TI - What's in store for medical students? Awareness and utilization of expert nutrition guidelines among medical school preceptors. AB - BACKGROUND: Instruction of physicians and other health professionals in medical nutrition sciences is among the expert recommendations to promote population health and reduce risks for cancer and other major causes of morbidity and mortality in the population. However, formal training in nutrition in United States medical schools is still lacking compared to the gains in basic and applied medical nutrition sciences. We sought to understand the awareness and current utilization of expert nutrition recommendations and practice guidelines among medical student faculty preceptors. METHODS: We surveyed the teaching faculty who precept for first-, second-, and third-year medical students in two required courses at Boston University. The instrument queried preceptor awareness and current utilization of expert nutrition recommendations, nutritional management practice guidelines, as well as faculty-student interactions regarding patient nutritional education and counseling. RESULTS: Of 187 faculty surveyed, 139 (74%) responded. Faculty reported using 2.3 expert guideline sources (N = 111; SD = 1.8; range = 0-8) but 83% had considered only one or no sources or did not remember what guidelines they had used. Eighty-four percent of preceptors expected students to routinely discuss nutritional practices with patients and/or their families; however, less than half of preceptors routinely provided feedback to students on patient nutritional education or counseling strategies. CONCLUSION: Our findings suggest gaps in faculty awareness and utilization of expert nutrition recommendations and practice guidelines relating to cancer and other chronic disease-risk reduction and population health promotion, underscoring the need for improvements in faculty and medical student training in basic and applied medical nutrition sciences. PMID- 15351543 TI - Dietary fat intake and lipid profiles of Iranian adolescents: Isfahan Healthy Heart Program--Heart Health Promotion from Childhood. AB - OBJECTIVES: To assess the serum lipid profiles of Iranian adolescents and their correlation with dietary fat intake and to evaluate the knowledge, attitude, and practice (KAP) of students, parents, and school staff. METHODS: The subjects of this cross-sectional study were 2000 students (1000 girls and 1000 boys), ages 11 18 years, selected by multistage random sampling, and one of their parents (2000 subjects), as well as 500 school staff in urban and rural areas of two provinces in Iran (one for further interventions and the other for reference). The data were obtained by questionnaires, anthropometric measurements, 3-day food record form, and a 20-item food frequency questionnaire (FFQ). All serum lipids were determined in the same laboratory. RESULTS: Although the percentage of fat intake (21.2 +/- 0.4%) among the adolescents was within the recommended daily allowance (RDA < or = 30%), in most cases, the percentiles of serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and triglyceride (TG) were significantly higher and the percentiles of high-density lipoprotein cholesterol (HDL-C) were lower than standard values according to Lipid Research Clinics (LRC) data; for example, the mean TC values for girls in the 11- to 14- and 15- to 18 year age groups were significantly higher than LRC standard values (169 and 172 vs. 160 and 159 mg/dl, respectively, P < 0.05). This difference was also significant in boys (167 and 168 vs. 160 and 153 mg/dl, respectively) at the P < 0.05 level. A significant linear association was shown between adolescents' dyslipidemia and the frequency of intake of hydrogenated fat, fast foods, cheese puffs, and potato chips (P < 0.05). Although the protein intake was lower than the RDA (13.4 +/- 0.9% vs. 15%, P < 0.05), because of the highly prevalent consumption of fatty lamb meat, the frequency of red meat intake had a direct association with dyslipidemia (P < 0.05). CONCLUSION: The improper intake of high amounts of saturated fat and the observed serum lipid profile of Iranian adolescents are likely placing them at increased risk for cardiovascular disease (CVD) and necessitate developing guidelines and community-based interventions. PMID- 15351544 TI - Individual susceptibility to hexavalent chromium of workers of shoe, hide, and leather industries. Immunological pattern of HLA-B8, DR3-positive subjects. AB - BACKGROUND: This study was designed to examine the effects of hexavalent chromium [Cr(VI)] on the immunological pattern of shoe, hide, and leather industry workers, moving from the hypothesis that some haplotypes (HLA-B8,DR3) can be important hidden risk cofactors. METHODS: Workplaces of 20 firms were monitored for total and respirable dusts and for total and hexavalent chromium. Cr(VI) on materials was also measured. Assay of chromium levels in blood and urine of 44 serological human leukocytes antigen (HLA)-typed workers (20 exposed, 15 HLA B8,DR3-negative/5-positive and 24 non-exposed, 18 HLA-B8,DR3-negative/6-positive subjects) was performed by atomic absorption, and lymphocyte subsets (FACS analysis), mitogen-mediate lympho-proliferation ([3H]thymidine incorporation), cytokine levels (ELISA), natural killer (NK) cytotoxic activity (51Cr-release assay) were determined. RESULTS: The environmental parameter levels are lower than threshold limit value-time-weighted average (TLV-TWA); in the materials, the Cr(VI) values exceeded the levels allowed. The peripheral blood mononuclear cells (PBMC) proliferation and the T-helper1 (TH1) cytokine pattern of subjects chronically exposed were significantly raised; addition in vitro of Cr(VI) further stimulated these parameters and in general the entire TH1 system and NK activity. The TH2 system was unaltered. In the HLA-B8,DR3-positive workers, immunologically "low responders", the addition of Cr(VI) in vitro caused a further reduction of the considered parameters in the exposed subjects with a dramatic deficit of the TH1 system. CONCLUSIONS: Results indicate the unsuitability of TLV-TWA as a line of demarcation between safe and dangerous Cr(VI) concentrations and the importance of individual genetic susceptibility for occupational and preventative medicine. In particular, the presence of the HLA B8,DR3 alleles can represent an important cofactor of immunotoxic susceptibility consequent to chronic low-dose Cr(VI) exposure. PMID- 15351545 TI - "Bulletproof skeptics in life's jungle": which self-exempting beliefs about smoking most predict lack of progression towards quitting? AB - OBJECTIVE: To determine the prevalence, correlates, and predictive value for intention to quit of 18 commonly expressed self-exempting beliefs about smoking among smokers and recent quitters, some 20 years after intensive tobacco control commenced in Australia. METHOD: National telephone survey of randomly selected 802 adults (685 smokers; 117 recent quitters). Main outcome measures. Level of agreement or disagreement with 18 self-exempting beliefs about smoking and intention to quit. RESULTS: Four coherent categories of self-exempting beliefs are widely held by smokers ("bulletproof", "skeptic", "jungle", and "worth it"). Smokers who hold self-exempting beliefs are more likely to be aged over 50, smoke more than 15 cigarettes per day, have less than 12 years of schooling, and be in the precontemplation stage of change. All scales had some relationship with progress towards quitting. In particular, "worth it" beliefs are powerful independent predictors of smokers not planning to quit. CONCLUSIONS: Some self exempting beliefs seem to act as a shield for smokers, giving them false reassurance and allowing them to avoid thinking deeply about the importance of quitting. This is particularly true of "worth it" beliefs. The prevalence of such beliefs may suggest confusion about smoking being a risk rather than a probable cause of illness. Creative approaches to increasing the saliency of the costs of smoking may be fruitful. PMID- 15351546 TI - Successful dietary changes in a cardiovascular risk reduction intervention are differentially predicted by biopsychosocial characteristics. AB - BACKGROUND: Psychosocial factors have demonstrated associations with both prognosis of patients with cardiovascular disease (CVD) as well as the development of CVD. Psychosocial distress may influence health behaviors (such as eating behavior) that increase cardiovascular risk. METHOD: The purpose of this research was to evaluate the role of psychosocial distress on intake of high-fat snacks and fruits and vegetables in 740 men and women, identified as high risk for cardiovascular disease, who took part in a 1-year cardiovascular risk factor reduction program. RESULTS: Participants who were more successful at decreasing or staying at an optimal level of high-fat food intake had lower levels of baseline total psychosocial distress and worry and decreases in total psychosocial distress. A greater success rate at increasing fruit and vegetable intake or staying at an optimal level of fruit and vegetable intake was associated with changes in Type A and competing behaviors. CONCLUSIONS: Results from these analyses suggest that baseline levels and changes in psychosocial variables, particularly decreased psychosocial distress, were associated with improvements in healthful eating behaviors. Further work may provide information that could be useful in tailoring health behavior interventions to specific subgroups based on psychosocial characteristics, initial health behaviors, and demographic characteristics. PMID- 15351547 TI - Level of knowledge and attitude of primary care physicians in Eastern Anatolian cities in relation to child abuse and neglect. AB - BACKGROUND: Child abuse and neglect is not a new phenomenon. It has always existed in every nation and culture in one form or another. The aim of this study was to determine the level of knowledge and attitude toward child abuse and neglect of primary care physicians serving in eastern Turkish cities. METHODS: The study was done in four Eastern Turkish cities involving Elazig, Tunceli, Malatya, and Bingol. All the primary care physicians serving in these cities were contacted. Of 266 primary care physicians, 175 replied (65.8% reply rate) and participated in the study. RESULTS: Most of the participated primary care physicians did not have adequate knowledge about physical assessment of suspected child abuse and neglect. Those physicians serving in primary care centers had better knowledge than those serving in Mother-Child Health and Family Planning Centers. CONCLUSIONS: Results from this study indicate that primary care physicians working in the eastern cities of Turkey do not have adequate knowledge and proper attitudes toward the identification and reporting of suspected child abuse. There is a need for educational programs to primary care physicians and other health personals working with child abuse and neglect to increase their knowledge and skills in detection, assessment, reporting, treatment, and prevention of child abuse and neglect. PMID- 15351548 TI - Validating outdoor workers' self-report of sun protection. AB - BACKGROUND: Few studies assessing sun safety behaviors have used validated survey instruments. Using an observational strategy, this study measured the validity of self-report of occupational sun protection behaviors. METHODS: Postal workers (N = 1,036) self-reported their frequency of protective clothing and sunglasses use during the past 5 workdays on Likert scales ranging from "Never" to "Always". Responses were dichotomized by Always vs. all other responses. Data collectors observed these same behaviors during mail delivery several days before survey administration. Agreement between the two measures was computed. RESULTS: Self report for all items was significantly related to observed behaviors, with Kappa values in the good to excellent range (Kappas = 0.51-0.83). CONCLUSIONS: For outdoor postal workers, survey items assessing recent occupational use of sun protective items had high levels of validity. PMID- 15351549 TI - Factors associated with the acceptance of an AIDS vaccine: an exploratory study. AB - BACKGROUND: Behavioral issues relative to acceptance of an HIV vaccine have been understudied. This exploratory study identified factors that may preclude people from accepting an AIDS vaccine when one is developed and approved for use. METHODS: A cross-sectional survey of 278 adults, from different at-risk populations, was conducted. Sixteen items assessed participants' concerns about receiving an AIDS vaccine if one was approved for use. Data reduction was achieved by principal components analysis, using an oblique rotation. Reliability and construct validity of subscales were determined. Finally, logistic regression models were used to identify demographic correlates of each precluding factor. RESULTS: A four-factor solution accounted for 59.5% of the variance. Three factors achieved satisfactory inter-item reliability: labeled as (1) personal issues, (2) product concerns, and (3) stigma-associated worry. Findings suggest that the three factors had construct validity. Each factor was associated with a unique demographic profile and two factors were associated with self-reported hepatitis B vaccination. CONCLUSION: Subsequent studies should use these exploratory findings (and possibly the measurement instrument) to tailor intervention programs aimed at promoting an AIDS vaccine. Because these studies will be time consuming, prudence dictates the initiation of further behavioral research before an AIDS vaccine is developed and approved. PMID- 15351550 TI - The changing age of HIV: sexual risk among older African American women living in rural communities. AB - BACKGROUND: African American women aged 50 and older are disproportionately affected by the HIV/AIDS epidemic. Despite African Americans making up 11% of all older women in the United States, in 2001, they accounted for more than 50% of AIDS cases among older women and more than 65% of HIV cases among older women. Using the AIDS Risk Reduction Model as a conceptual framework, this study investigated HIV risk behavior among older rural African American women (mean age = 58 years). METHODS: A cross-sectional survey was conducted (n = 181) in three rural counties in South Carolina. RESULTS: Most (67%) of the women had at least one sex partner in the past five years, and of these, more than half (59.5%) reported at least one sexual risk behavior. High-risk behavior was associated with less education, lower condom use self-efficacy, more peers who discussed HIV related risk behavior, and less comfort communicating with partners about sex. CONCLUSIONS: A significant proportion of older African American women living in rural counties are at increased risk for HIV infection, suggesting an urgent need for HIV prevention efforts to target this population. PMID- 15351551 TI - Diet and physical activity counseling during ambulatory care visits in the United States. AB - BACKGROUND: Research is limited regarding national patterns of behavioral counseling during ambulatory care. We examined time trends and independent correlates of diet and physical activity counseling for American adults with an elevated cardiovascular risk during their outpatient visits. METHODS: The National Ambulatory Medical Care Survey (NAMCS) and National Hospital Ambulatory Medical Care Survey (NHAMCS) provided 1992-2000 national estimates of counseling practices in private physician offices and hospital outpatient departments. RESULTS: Rates of diet and physical activity counseling among visits by at-risk adults exhibited a modest ascending trend from 1992 to 2000, with the biggest growth found between 1996 and 1997. Throughout the 1990s, however, diet counseling was provided in <45% and physical activity counseling in < or = 30% of visits by adults with hyperlipidemia, hypertension, obesity, or diabetes mellitus. Lower likelihood of either counseling was significantly associated with patients who were > or = 75 years of age, seen by generalists, and those with fewer risk factors. Also, diet counseling was less frequently provided during visits by whites vs. ethnic minorities and by men vs. women. CONCLUSIONS: Despite available national guidelines, diet and physical activity counseling remain below expectations during outpatient visits by adults with an elevated cardiovascular risk. Given recent trends, immediate, satisfactory improvement is unlikely without future innovative interventions. PMID- 15351552 TI - Exercise programs for older men: mode and intensity to induce the highest possible health-related benefits. AB - BACKGROUND: Although health-related benefits of fitness training in older men are well established, it is not clear yet which mode and intensity of a exercise program is most effective. This study addresses whether the combination of endurance (ED) and resistance training in older men have supplementary health related benefits in addition to profits attained through endurance training alone. Additionally, effects of moderate- and low-intensity resistance training are compared. METHODS: Men, 55-75 years of age, were randomly assigned to a control group (N = 13) or one of three exercise groups (20 weeks, two to three times per week): endurance plus moderate resistance (MR) training (N = 22), endurance plus low resistance (LR) training (N = 22) and endurance training only (N = 22). Cardiovascular (CV) risk factors, muscular fitness and postural control were assessed before and after training. RESULTS: All exercise groups revealed significant (P < 0.05) improvements in resting heart rate, work capacity and recovery, waist girth, insulin response and knee-extensor strength with no differences among groups. Body composition, resting metabolic rate (RMR), VO2peak and postural control did not change in exercise groups. CONCLUSION: In older men, a fitness program consisting of 20 weeks endurance training combined with resistance training is equally effective as endurance training alone. Moderate vs. low resistance training added to endurance training yields similar health related benefits. PMID- 15351553 TI - School-based opportunities for physical activity in West Virginia public schools. AB - BACKGROUND: Schools have the unique potential to provide numerous opportunities for promoting physical activity. This article describes findings from a statewide survey of opportunities for physical activity in West Virginia (WV) schools. The purpose was to provide baseline data for two of the WV Healthy People 2010 objectives related to schools and youth to identify priorities for action. METHOD: Survey questions were adapted from the 2000 School Health Policies and Programs Study (SHPPS), conducted by CDC. Random stratified sampling across school level and size resulted in a final sample of 296 elementary schools, 146 middle and junior high schools, and 124 high schools (total = 566). The overall response rate was 73%. RESULTS: Eleven percent of elementary, 2% of middle or junior high, and 31% of senior high schools met the SHPP's criterion of providing daily physical education. Ninety-four percent of elementary schools reported offering daily recess. Overall, 42.3% of schools provided student and community access to indoor facilities outside of normal school hours, while 80.7% of schools provided access to outdoor facilities beyond normal school hours. CONCLUSIONS: Survey results are being used to target increased physical education in elementary schools and increased opportunities beyond physical education at all school levels. PMID- 15351554 TI - An index to measure the association between dietary patterns and coronary heart disease risk factors: findings from two Italian studies. AB - BACKGROUND: To examine the association between established coronary heart disease (CHD) risk factors and eating patterns by creating a dietary index utilizing a limited amount of food frequency questionnaire data on specified indicator foods. METHODS: Selected data from the Italian Risk Factors and Life Expectancy (RIFLE) project collected between 1978-1979 and 1983-1984 on 7665 men and women, aged 20 59 years, were used to compute a dietary index summarizing the relative proportion of fatty to non-fatty foods. The association between this index and systolic and diastolic blood pressure (SBP, DBP), body mass index, total serum cholesterol (TC), and blood glucose, controlling for potential confounders was then tested using multiple linear regression. RESULTS: Among men, a positive association was found between the index and all five CHD risk factors (P < 0.01). Among women, the associations were weaker, becoming statistically significant only for TC and glucose. CONCLUSIONS: Our food index was able to detect relationships between diet and traditional CHD risk factors. A tool such as this, which requires only a limited number of key indicator food items, might enhance the use of existing food frequency questionnaires and also streamline the process of collecting new dietary information. PMID- 15351555 TI - Current and new strategies in immunotherapy for superficial bladder cancer. PMID- 15351556 TI - Systemic inflammatory response syndrome in open versus laparoscopic adrenalectomy. AB - OBJECTIVES: To compare the surgical invasiveness of laparoscopic adrenalectomy with that of open surgery when the invasiveness is evaluated by the incidence of systemic inflammatory response syndrome (SIRS). METHODS: Patients who underwent open adrenalectomy (35 patients) and those who underwent transperitoneal laparoscopic adrenalectomy (35 patients) were included in this study. We retrospectively investigated the incidence and duration of SIRS in the two groups. Of the 70 patients, 39 had primary aldosteronism and 31 had clinically nonfunctioning adrenal tumors. RESULTS: The incidence of SIRS was 51.4% (18 patients) in the open group and 31.4% (11 patients) in the laparoscopic group (P = 0.14, Fisher's exact probability test). The mean duration of SIRS was 1.9 days in the open group and 1.2 days in the laparoscopic group (P = 0.04, unpaired t test). Surgical site infection, which occurred in 2 patients in the open group (5.7%) and 3 patients (8.5%) in the laparoscopic group, was the only infectious complication. No patient had any severe or life-threatening complications. CONCLUSIONS: Laparoscopic adrenalectomy seems less invasive than open surgery, when the magnitude of the invasiveness is evaluated by the incidence of SIRS. PMID- 15351557 TI - Percutaneous nephrolithotomy in treatment of large stones within horseshoe kidneys. AB - OBJECTIVES: To present our experience with percutaneous nephrolithotomy (PCNL) in the treatment of large stones within horseshoe kidneys. METHODS: The study included 34 patients with 45 stone-bearing horseshoe kidneys treated by PCNL in one center. The indications for PCNL were large stone burden (greater than 2 cm) in 10 kidneys (22.2%), multiple complex stones in 22 (48.9%), staghorn stones in 3 (6.7%), and failed extracorporeal shock wave lithotripsy (ESWL) in 10 kidneys (22.2%). All PCNL procedures were performed in a standard one-session technique with fluoroscopic guidance. Procedures were evaluated for intraoperative and postoperative complications, use of auxiliary procedures (second-look PCNL, ureteroscopy, and ESWL), and length of hospital stay. Moreover, the stone-free rate was calculated at discharge from the hospital and 3 months later. In addition, we assessed the need for secondary intervention during a mean follow-up of 75 +/- 16 months (range 6 to 108). RESULTS: The stone burden ranged from 264 to 2408 mm(2) (mean 664 +/- 153). Access to the horseshoe kidneys was supracostal in 10 (22.2%) and subcostal in 35 kidneys (77.8%). A single tract was used in 37 (82%), and two tracts were created in the remaining 8 kidneys (18%). Major complications were seen in six PCNL procedures (13.3%) and included significant hematuria requiring blood transfusion in three, septicemia in one, ureteral obstruction in one, and colonic injury in one. All complications were successfully treated. Auxiliary procedures were required in 12 patients (35.3%) and included ureteroscopy in 1, second-look PCNL in 3, and ESWL to fragment residual caliceal stones in 8 patients. The mean hospital stay was 4 +/- 1.9 days (range 3 to 12). The stone-free rate was 82% at discharge and 89% at 3 months. During the course of follow-up, 19 patients (56%) developed stone recurrence and were successfully treated with ESWL in 8 and PCNL in 11. CONCLUSIONS: PCNL is a safe and effective procedure in the treatment of large stones within horseshoe kidneys. PMID- 15351558 TI - Clinical utility of dual active deflection flexible ureteroscope during upper tract ureteropyeloscopy. AB - OBJECTIVES: To evaluate the clinical utility of a dual active deflection ACMI DUR 8 Elite ureteroscope in a referral endourology practice. METHODS: Retrospective chart review was performed on 54 consecutive patients who underwent flexible ureteroscopy by a single surgeon (S.Y.N.) from February to July 2003. Cases in which standard flexible ureteroscopes alone could complete the procedure, cases in which standard flexible ureteroscopy could not complete the procedure and the DUR-8 Elite ureteroscope did, and cases in which both ureteroscopes failed to complete the procedure were analyzed. RESULTS: A total of 54 procedures were performed on 37 patients. Three cases were not analyzed because they were distal ureter procedures. Of the remaining 51 procedures, 6 were removed from analysis because they were second-look procedures. When classified by diagnosis, 27 patients had stones (79.4%), 5 had cancer (14.7%), and 1 had hematuria (2.9%). The global success rate was 91.1%. The average use rate of the DUR-8 was 28.9%, and the success rate using the DUR-8 Elite was 69.2% in those cases in which it was necessary. Of the 13 cases in which the DUR-8 was used, 61.5% were for lower pole pathologic findings. The DUR Elite use and success rate in the lower pole was 57.1% and 75%, respectively. A statistically significant association was found between the diagnosis and procedure location (P = 0.00128). CONCLUSIONS: Our preliminary data indicate that the dual deflecting DUR-8 Elite ureteroscope may be helpful in cases in which the single deflection flexible instruments fail to access and treat upper urinary tract pathologic findings. PMID- 15351559 TI - Comparison of tipless and helical baskets in an in vitro ureteral model. AB - OBJECTIVES: To compare the rapidity of stone capture and removal by six tipless and four helical baskets in a ureteral model. Expeditious ureteral stone extraction is reliant on the basket size, configuration, and radial dilation force. METHODS: A 5-mm feline calculus was placed in a simulated model of the human ureter at a distance of 7.5 cm from the orifice, and saline irrigant instilled. A 6/7.5F Wolf semirigid ureteroscope was used for access and visualization of the stone. Operators were randomized to start stone extraction with 1 of 10 stone baskets, and three extraction attempts were conducted with each device. Six experienced operators tested each device. The tipless baskets tested were the ACMI Sur-Catch NT 3.0F, Bard Dimension 3.0F, Boston Scientific Zerotip 3.0F, Cook N-Circle 3.0F, Cook Delta 2.4F, and Sacred Heart Halo 1.9F. The helical baskets tested were the Cook N-Force 3.2F, Microvasive Gemini 3.0F, ACMI Sur-Catch Helical 3.0F, and Sacred Heart Hercules 3.0F. The time to extraction in seconds was recorded. RESULTS: No statistically significant difference was observed between the tipless (14 +/- 7 seconds) or helical (12.8 +/- 6.6 seconds, P = 0.21) basket groups. All baskets were able to retrieve the ureteral stones successfully. The most efficient basket was the Cook N-Circle with an average time of 8.7 seconds to stone extraction compared with the least efficient basket, the ACMI Sur-Catch NT, at 15.8 seconds. CONCLUSIONS: The Cook N Circle basket provided the most expeditious means to stone extraction in a ureteral model. PMID- 15351562 TI - Is there a role for small-diameter ureteral access sheaths? Impact on irrigant flow and intrapelvic pressures. AB - OBJECTIVES: To evaluate irrigant flows and intrapelvic pressures with small diameter access sheaths. Ureteral access sheaths improve irrigant flow and decrease intrarenal pelvic pressures during flexible ureteroscopy. However, no comparisons of individual sheaths have been conducted. Previous studies have demonstrated more favorable results with the 12F sheath than with the 10F sheath. METHODS: Ureteral access sheaths were tested ex vivo in porcine kidneys. An 18F angiocatheter was placed in the renal pelvis and connected to a Hewlett Packard Gauss Pressure transducer. Irrigant was maintained at 100 mm Hg pressure. Irrigant flow and intrapelvic pressures were measured with three flexible ureteroscopes at baseline and using each of four 10F sheaths, with the sheaths positioned in the middle ureter and the ureteroscopes positioned in the renal pelvis. The pressure at which irrigant efflux through the sheath occurred and the rate of irrigant efflux through the access sheath were measured. RESULTS: Intrapelvic pressures measured greater than 40 mm Hg, and irrigant flows remained at less than 15 mL/min when the Olympus URF-P3 and Storz 11274AAU flexible ureteroscopes were tested with all four sheaths. The intrapelvic pressures, irrigant inflow, and irrigant efflux with the Wolf 7325.172 (7.5F) flexible ureteroscope were optimized in combination with the Cook Peelaway 10F and Applied Access 10F sheaths. CONCLUSIONS: Small ureteral access sheaths should be used only with the Wolf 7325.172 flexible ureteroscope. The Cook Peelaway (10F) and Applied Access (10F) sheaths offered the greatest increase in irrigant flow and decrease in intrapelvic pressures. PMID- 15351565 TI - Electrocauterization of bleeding points for percutaneous nephrolithotomy. AB - OBJECTIVES: To perform a retrospective study to assess the efficacy and safety of electrocauterization of bleeding points after stone extraction using the data from 249 percutaneous nephrolithotomies. METHODS: A total of 341 percutaneous nephrolithotomies in 324 patients were performed at the Chiayi Christian Hospital from July 2000 to July 2003. Electrocauterization of bleeding points with an elongated electrode probe was performed in 249 patients. The age, height, weight, preoperative hemoglobin level, stone burden, operating time, stone free rate, length of postoperative hospital stay, postoperative urinary tract infection rate, and blood transfusion rate were recorded by retrospective chart review. RESULTS: No statistically significant differences in age, height, weight, stone burden, operating time, stone free rate, or length of postoperative hospital stay were found between patients with or without electrocauterization. No increase occurred in the postoperative urinary tract infection rate in patients who received electrocauterization, and these patients had a statistically significant decrease in the transfusion rate. No nephrostomy tube was inserted at the completion of surgery in 84 (33.7%) of the 249 operations in which electrocauterization was performed. CONCLUSIONS: Electrocauterization of the bleeding points at the end of percutaneous renal surgery decreases the blood transfusion rate without causing an increase in morbidity. This procedure is safe and effective and may make more patients suitable for tubeless modification. PMID- 15351567 TI - Stripping technique for endoscopic management of distal ureter during nephroureterectomy: experience of 32 procedures. AB - OBJECTIVES: To assess the results of the stripping technique for endoscopic management of the distal ureter during nephroureterectomy. METHODS: Since 1995, 32 patients have undergone open nephroureterectomy involving stripping of the distal ureter for upper urinary tract carcinoma. After endoscopic placement of a Chevassu ureteral catheter, nephrectomy was performed using a lumbotomy approach. The catheter tip was tied to the top of the ureteral extremity. At the end of the intervention, the catheter was pulled out transurethrally to intussuscept the distal ureter. Endoscopic resection through the bladder muscular wall was performed around the everted ureteral orifice, then the distal ureter was stripped and removed with a bladder cuff. RESULTS: The mean operative time was 180 minutes, and the mean blood loss was 220 mL. The stripping failed in 6 patients (18.7%), requiring an iliac incision. The reasons for failure were a retained ureter in 5 cases and ureteral breakage in 1 case. The mean duration of urethral catheterization was 5 days, and the mean hospital stay was 8 days. The complication rate was 3.1%. At a median follow-up of 35.2 months, 9 patients (28.1%) had experienced recurrence: 6 with superficial bladder tumors, 2 with renal fossa recurrence, and 1 with asynchronous liver metastases. The disease specific mortality rate was 9.4%. CONCLUSIONS: These results confirm that the stripping technique for endoscopic management of the distal ureter during open nephroureterectomy is a simple and safe procedure. PMID- 15351569 TI - Preoperative renal tumor evaluation by three-dimensional magnetic resonance imaging: Staging and detection of multifocality. AB - OBJECTIVES: To evaluate, retrospectively, the staging accuracy of three dimensional magnetic resonance imaging (3D-MRI) in our institution as a prelude to a prospective comparison of 3D-MRI and 3D computed tomography (CT) for preoperative planning of partial nephrectomy. In recent years, the use of 3D-CT for preoperative evaluation and surgical planning in patients undergoing nephron sparing surgery has gained considerable popularity. METHODS: The images of 26 consecutive patients evaluated by 3D-MRI as part of the preoperative imaging studies for renal tumor were evaluated retrospectively and compared with the surgical pathologic findings to evaluate the ability of 3D-MRI to predict tumor multifocality, tumor stage, collecting system invasion, and venous invasion. RESULTS: 3D-MRI accurately predicted tumor multifocality in 1 of 2 cases. Imaging identified five of seven multifocal lesions. Two subcentimeter lesions were missed. Preoperative staging was correct in 29 of 30 lesions (97% accuracy). One T3b tumor was incorrectly staged as T2. Venous invasion was identified in 2 of 3 cases (67% sensitivity), but no false-positive results were seen. 3D-MRI had 100% sensitivity and 88% specificity in the prediction of collecting system invasion. CONCLUSIONS: The staging accuracy of 3D-MRI appears to be quite good. Given the accuracy of this technique, along with the popularity of 3D imaging before renal surgery, these results provide the impetus for a future study directly comparing 3D-CT with 3D-MRI in the capacity of surgical preoperative planning. PMID- 15351570 TI - Laparoscopic versus open partial nephrectomy. AB - OBJECTIVES: To compare, retrospectively, the results of laparoscopic partial nephrectomy (LPN) to open partial nephrectomy (OPN) using a tumor size-matched cohort of patients. Limited data are available comparing LPN to OPN in the treatment of small renal tumors. METHODS: Between September 2000 and September 2003, 27 LPNs and 22 OPNs were performed to treat renal masses less than 4 cm. Patient demographics and tumor location and size (2.4 +/- 1.0 cm versus 2.9 +/- 0.9 cm, respectively; P = not statistically significant) were similar between the LPN and OPN groups. RESULTS: Although the mean operative time was longer in the LPN than in the OPN group (210 +/- 76 minutes versus 144 +/- 24 minutes; P <0.001), the blood loss was comparable between the two groups (250 +/- 250 mL versus 334 +/- 343 mL; P = not statistically significant). No blood transfusions were performed in either group. The hospital stay was significantly reduced after LPN compared with after OPN (2.9 +/- 1.5 days versus 6.4 +/- 1.8 days; P <0.0002), and the postoperative parenteral narcotic requirements were lower in the LPN group (mean morphine equivalent 43 +/- 62 mg versus 187 +/- 71 mg; P <0.02). Three complications occurred in each group. With LPN, no patient had positive margins or tumor recurrence. Also, direct financial analysis demonstrated lower total hospital costs after LPN (4839 dollars+/- 1551 dollars versus 6297 dollars+/- 2972 dollars; P <0.05). CONCLUSIONS: LPN confers several benefits over OPN concerning patient convalescence and costs, despite prolonged resection times at our current phase of the learning curve. Long-term results on cancer control in patients treated with LPN continue to be assessed. PMID- 15351571 TI - Long-term survival of patients with unilateral sporadic multifocal renal cell carcinoma according to histologic subtype compared with patients with solitary tumors after radical nephrectomy. AB - OBJECTIVES: To determine the pathologic features, including histologic subtype and tumor size, associated with multifocal renal cell carcinoma (RCC) and the impact of multifocality on survival after radical nephrectomy, about which controversy exists. METHODS: We studied 2373 patients who underwent radical nephrectomy for RCC from 1970 to 2000. Histologic subtype, stage (2003 TNM), nuclear grade, tumor size, and multifocality, defined as the presence of more than one ipsilateral RCC tumor of the same histologic subtype, were evaluated. Associations of multifocality with ipsilateral and contralateral recurrence and death from RCC were evaluated using Cox proportional hazards models. RESULTS: The incidence of sporadic histologically concordant multifocality was greater in papillary RCC (29 of 266; 10.9%) compared with clear cell RCC (40 of 1934; 2.0%; P <0.001) and chromophobe RCC (2 of 104; 1.9%; P = 0.005). Patients with solitary clear cell and papillary RCC had larger tumors compared with multifocal clear cell and papillary RCC (P <0.001 and P = 0.024, respectively). Patients with multifocal clear cell RCC were more likely to have a contralateral recurrence than were patients with solitary clear cell RCC (risk ratio 2.91, P = 0.142). Multifocality was not significantly associated with ipsilateral recurrence or death from RCC in patients with clear cell or papillary RCC. CONCLUSIONS: The incidence of multifocality was significantly greater among patients with papillary RCC than among patients with clear cell or chromophobe RCC. Patients with multifocal clear cell RCC were more likely to experience a contralateral recurrence. Multifocality was not significantly associated with ipsilateral recurrence or death from RCC. In addition, multifocality was not associated with larger and higher stage tumors, as previously reported. PMID- 15351572 TI - Loss of calponin h1 in renal angiomyolipoma correlates with aggressive clinical behavior. AB - OBJECTIVES: To investigate whether any immunohistochemical differences exist between two types of renal angiomyolipomas (AMLs). Renal AMLs are generally considered to be benign in nature. However, a few of these tumors have been reported to involve other organs. METHODS: Tissue specimens from 3 cases of clinically aggressive and 9 cases of clinically indolent renal AMLs were examined histologically and immunohistochemically using antibodies against calponin h1 and alpha-smooth muscle actin. Immunostaining was evaluated semiqualitatively as no staining to strong staining. RESULTS: The tumor cells in the myomatous component of the aggressive type did not show any reaction to the antibody to calponin h1 protein, but they showed strong immunoreactions with the antibody to the alpha smooth muscle actin antigen. All cases of the nonaggressive AMLs demonstrated strong immunoreactions with both antibodies used. CONCLUSIONS: Our initial results in a small series of cases suggest a potential molecular difference between the aggressive and nonaggressive type of AMLs. Loss of calponin h1 from the tumor cells of the aggressive type might be related to their pathologically invasive features, and this aggressive type might be categorized into an intermediate type between the benign and malignant types. PMID- 15351573 TI - Significance of heat shock protein-27 expression in patients with renal cell carcinoma. AB - OBJECTIVES: To investigate heat shock protein (HSP)-27 expression in patients with renal cell carcinoma (RCC) and examine its biologic significance. HSPs were first defined as proteins induced by heat shock and other environmental and pathophysiologic stresses. They are implicated in protein-protein interactions and are thought to play an important role in cancer. The expression of HSP-27 has been demonstrated in some human tumors. METHODS: The expression of HSP-27 was studied in tumor and normal parenchyma tissue specimens from 76 patients with RCC by immunohistochemistry. The findings were correlated with clinical stage, lymph node metastasis, histologic grade, and survival. RESULTS: Of the 76 RCC tissue specimens studied, the presence of HSP-27 was demonstrated in 73 (96%). The expression was low in 10 patients (14%), intermediate in 38 (50%), and high in 25 (33%). HSP-27 expression was greater in RCC tissue compared with adjacent noncancerous renal tissue (P <0.001). An inverse relationship was found between tumor stage and HSP-27 expression (r = -0.281, P = 0.016). However, no statistically significant difference was observed in progression-free survival with respect to HSP-27 expression. No relationship was found between HSP-27 expression and tumor grade, lymph node metastasis, distant metastasis, or cause specific survival. CONCLUSIONS: Our data suggest that HSP-27 expression is not a powerful and statistically significant prognostic indicator for disease-free survival for patients with RCC. PMID- 15351574 TI - Gemcitabine and carboplatin combination as first-line treatment in elderly patients and those unfit for cisplatin-based chemotherapy with advanced bladder carcinoma: Phase II study of the Hellenic Co-operative Oncology Group. AB - OBJECTIVES: To evaluate, in a multicenter Phase II study, the safety and efficacy of the combination of gemcitabine and carboplatin, as first-line treatment in elderly and unfit patients with advanced bladder carcinoma. The toxicity of platinum-based chemotherapy combinations represents a common problem for elderly or unfit patients with advanced bladder carcinoma. METHODS: Patients with previously untreated inoperable or metastatic bladder carcinoma and an Eastern Cooperative Oncology Group performance status greater than 2, age older than 75 years, or creatinine clearance of less than 50 mL/min were treated with carboplatin area under the curve 4 on day 1 and gemcitabine 1000 mg/m(2) on days 1 and 8, every 21 days for a total of six cycles. RESULTS: A total of 56 patients (48 men and 8 women, median age 75 years) were enrolled. Of these patients, 46% had a performance status of 2 to 3, 68% had a creatinine clearance of less than 50 mL/min, and 59% had distant metastases. The overall response rate was 36% (95% confidence interval 23.4% to 49.6%), and an additional 14 patients had disease stabilization (25%, 95% confidence interval 14.4% to 38.4%). The median time to progression was 4.8 months, the median overall survival was 7.2 months, and the 1 year survival rate was 26%. Grade 3 or 4 toxicity included anemia (18%); thrombocytopenia (16%); neutropenia (27%), with two episodes of febrile neutropenia requiring hospitalization; diarrhea (2%); and fatigue (5.5%). Two toxic deaths occurred during the study. CONCLUSIONS: The combination of gemcitabine and carboplatin has some activity as first-line treatment of advanced bladder carcinoma in the elderly and those unfit for cisplatin-based chemotherapy, with manageable toxicity, and represents a reasonable choice for the treatment of such patients. PMID- 15351575 TI - Urinary conduit formation using retubularized bowel from continent urinary diversion or intestinal augmentations: I. A multi-institutional experience. AB - OBJECTIVES: To present our multi-institutional experience with retubularized bowel conduits. Some patients with augmentation cystoplasty, augmented rectal bladder, or continent urinary reservoirs require conversion to an ileal conduit for various reasons. This is generally accomplished by construction of a de novo ileal conduit. We retubularized the bowel segment used in the augmented bladder or the urinary reservoir to form the intestinal urinary conduit. METHODS: A total of 29 patients (mean age 42 years) underwent conduit formation using the augmentation patch of enterocystoplasty, the segment of cutaneous urinary diversion reservoir or orthotopic neobladder, or the augmentation patch of an augmented rectal bladder. Patients were followed up with history, physical examination, laboratory tests, and imaging studies. RESULTS: The follow-up ranged from 12 to 130 months (mean 42). One patient developed small bowel obstruction requiring laparotomy with lysis of adhesions. None of the other patients had bowel complications. Two patients developed ureteral obstruction with hydronephrosis. One had pyocystitis requiring simple cystectomy. One patient was lost to follow-up. CONCLUSIONS: Retubularization of previously used bowel segments to construct a urinary conduit is feasible and has several advantages. It is possible to perform this procedure after several types of reconstructive procedures in which bowel was incorporated into the urinary tract. PMID- 15351577 TI - Clinical outcome in patients with locally advanced bladder carcinoma treated with conservative multimodality therapy. AB - OBJECTIVES: To describe the outcome of patients with muscle-invasive bladder carcinoma treated with multimodality therapy in our institution from 1993 to 2002. METHODS: The charts of 60 patients with Stage T2-T4N0-N1M0 treated with transurethral resection of bladder tumor followed by a chemoradiotherapy combination were retrospectively reviewed. Of the 60 patients, 22 had received neoadjuvant chemotherapy (methotrexate, cisplatin, and vinblastine or methotrexate, adriamycin, cisplatin, and vinblastine) followed by concomitant chemoradiotherapy (weekly cisplatin/carboplatin or a cisplatin and 5-fluorouracil combination), and 38 had received concomitant chemoradiotherapy alone. Radiotherapy delivered a median dose of 45 Gy to the pelvis and 65 Gy to the bladder in a once-daily or twice-daily fractionation scheme. Follow-up evaluations included cystoscopy with biopsies at regular intervals. Salvage cystectomy was recommended in the case of local persistent tumor or bladder relapse. RESULTS: The median follow-up was 48.5 months (range 10 to 126). Of the 22 patients who received neoadjuvant chemotherapy, 18 (82%) had received two or more cycles; 51 (85%) of the 60 patients received the concomitant chemotherapy as planned. Radiotherapy was completed in 56 patients. Twenty-eight patients developed relapse either locally (14 did not achieve a complete local response after chemoradiotherapy and 6 had true local relapse during follow-up) or at distant sites. The actuarial 5-year disease-specific survival and freedom from local and distant relapse rate was 54% and 42%, respectively. The actuarial local control rate with an intact bladder was 56% at 5 years. When stratified according to stage and grade, patients with Stage T2-T3, grade 2 tumors had a statistically significantly better chance of remaining relapse free than did the others (P = 0.045). Salvage cystectomy (n = 11) for isolated local failure in this population achieved limited results. CONCLUSIONS: Transurethral resection of bladder tumor with this chemoradiotherapy combination achieved satisfactory results in this unfavorable population with invasive bladder carcinoma. PMID- 15351578 TI - Analysis of treatment for small cell cancer of the bladder and report of three cases. AB - OBJECTIVES: To present our experience with 3 patients with small cell cancer (SCC) of the bladder who were treated with different modalities and review the literature for patients undergoing primary chemoradiotherapy. SCC of the bladder is a rare tumor, with patients commonly presenting with metastatic disease. Surgery, radiotherapy, and chemotherapy, either alone or as part of combined therapy, have been used. Because of the rarity of this disease, no prospective studies evaluating the most effective treatment have been done. METHODS: The medical records of 3 patients diagnosed with SCC of the bladder at our institution were reviewed. Additionally, we reviewed published reports to identify all cases of SCC of the bladder treated with primary chemoradiotherapy. RESULTS: Three patients with SCC of the bladder were identified at our institution. A total of 23 patients with SCC of the bladder who were treated with primary chemoradiotherapy were identified: 22 in published reports and 1 at our institution. Patients presented with muscle-invasive disease (17%), extravesical disease only (26%), and metastatic disease (52%). Multiagent chemotherapy was administered to most patients. The reported median radiation dose was 6000 cGy. A total of 16 patients (70%) were alive at a median follow-up of 34 months. The median survival of patients had not yet been reached in this study at the last follow-up. We did not find any reports of SCC recurrence in the bladder, and the bladder was preserved in most patients (87%). CONCLUSIONS: SCC of the bladder should be viewed as a systemic disease, because most patients present with metastatic disease. Primary chemoradiotherapy appears to be an effective treatment modality. Prospective studies are needed to evaluate the optimal treatment further. PMID- 15351579 TI - Short-term effects of increased urine output on male bladder function and lower urinary tract symptoms. AB - OBJECTIVES: To determine whether the human bladder can also adapt to an increased physiologic load, because bladder dysfunction is considered to be an important factor in the development of lower urinary tract symptoms. Animal studies have shown that bladder function can be improved by increasing the urine output. METHODS: A total of 44 men between 55 and 75 years of age were asked to increase their daily fluid intake by 2 L for a 2-month period. The objective outcome measures were maximal urinary flow rate (Da Capo home uroflowmeter), maximal voided volume (frequency-volume chart), and average voided volume (frequency volume chart). The International Prostate Symptom Score and global perceived benefit of the intervention were recorded to assess the subjective effects of the intervention. RESULTS: The maximal flow rate increased by 13% (2.3 mL/s), the maximal voided volume increased by 23% (93.8 mL), and average voided volume increased by 25% (60.1 mL). Of the 44 participants, 56% reported an improvement in their lower urinary tract function, but the International Prostate Symptom Score increase was statistically significant at 1.2 point. CONCLUSIONS: The human bladder seems able to adapt to an increased load. Future randomized effectiveness studies with longer follow-up should be done to determine the upper limit of objective bladder adaptation. In addition, future studies should address the long term efficacy in the prevention of symptoms. PMID- 15351580 TI - Association between serum concentrations of micronutrients and lower urinary tract symptoms in older men in the Third National Health and Nutrition Examination Survey. AB - OBJECTIVES: To evaluate the association of serum micronutrients with lower urinary tract symptoms (LUTS). METHODS: We included 2497 men, 60 years old and older, who participated in the Third National Health and Nutrition Examination Survey from 1988 to 1994 and for whom serum concentrations of vitamins A, C, and E, carotenoids, and selenium had been measured previously. Cases were men with three or four of the following symptoms: nocturia, hesitancy, incomplete emptying, and weak stream, but who had never undergone noncancer prostate surgery. Controls were men without symptoms, who had never undergone noncancer prostate surgery. We adjusted for age and race in logistic regression models and used sampling weights. RESULTS: Serum concentrations of vitamin E (P = 0.03), lycopene (P = 0.06), and selenium (P = 0.03) were lower in men with LUTS compared with controls. Men in the top four quintiles of vitamin E, lycopene, and selenium had a nonstatistically significant 25% to 50% reduced odds of LUTS compared with men in the bottom quintile. Inverse associations were not seen for the other carotenoids or vitamin A. A high serum vitamin C concentration was associated with a lower odds of LUTS in current smokers, but with a nonstatistically significant greater odds in those who never smoked and in former smokers. CONCLUSIONS: Greater circulating concentrations of vitamin E, lycopene, and selenium, antioxidant micronutrients that are supported in published reports as protecting against prostate cancer, were observed also to be inversely associated with LUTS. The effect modification of the association with vitamin C by cigarette smoking warrants additional examination. PMID- 15351581 TI - Effects of a diet rich in phytoestrogens on prostate-specific antigen and sex hormones in men diagnosed with prostate cancer. AB - OBJECTIVES: To determine the effects of diets rich in soy and linseed compared with a control diet on biochemical markers of prostate cancer in men diagnosed with prostate cancer. METHODS: Twenty-nine men diagnosed with prostate cancer and scheduled to undergo a radical prostatectomy were randomized to one of three groups: soy (high phytoestrogen), soy and linseed (high phytoestrogen), or wheat (low phytoestrogen). A bread was specially manufactured to incorporate 50 g of heat-treated (HT) soy grits or 50 g of HT soy grits and 20 g of linseed as part of the study participant's daily diet. Baseline and preoperative levels of prostate-specific antigen (PSA), free PSA, testosterone, sex hormone-binding globulin, free androgen index, and dihydrotestosterone were measured. RESULTS: Statistically significant differences were detected between the HT soy grits group and the control wheat group for the percentage of change in total PSA ( 12.7% versus 40%, P = 0.02) and the percentage of change in free/total PSA ratio (27.4% versus -15.6%, P = 0.01); and between the HT soy grits group and the HT soy grits and linseed group for the percentage of change in free androgen index (16.4% versus -15.5%, P = 0.04) and the percentage of change in free/total PSA ratio (27.4% versus -10%, P = 0.007). CONCLUSIONS: The data from this study indicate that a daily diet containing four slices of a bread rich in HT soy grits favorably influences the PSA level and the free/total PSA ratio in patients with prostate cancer. This work provides some evidence to support epidemiologic studies claiming that male populations who consume high phytoestrogen diets have a reduced risk of prostate cancer development and progression. PMID- 15351582 TI - Preoperative neural network using combined magnetic resonance imaging variables, prostate-specific antigen, and Gleason score to predict positive surgical margins. AB - OBJECTIVES: To assess whether artificial neural network analysis (ANNA) predicts for positive surgical margins (PSMs) more effectively than logistic regression analysis (LRA) according to the combined use of the findings of pelvic coil magnetic resonance imaging (pMRI) and other preoperatively available tumor variables in patients with clinically organ-confined prostate cancer after radical prostatectomy. METHODS: A total of 205 patients with clinically localized prostate cancer, who underwent retropubic radical prostatectomy were evaluated. The predictive variables included clinical TNM stage, prostate-specific antigen (PSA) level, PSA density, biopsy Gleason score, percentage of cancer in biopsy specimens, and pMRI findings. The predicted outcome was PSMs. The patient data were randomly split into four cross-validation sets and used to develop and validate the ANNA and LRA models. For comparison, the area under the receiver operating characteristic curve was used. RESULTS: The overall PSM rate was 22% (n = 45). Using all input parameters, the accuracy of the ANNA and LRA was 84% and 75% for the prediction of PSMs, respectively. The area under the receiver operating characteristic curve of the ANNA (0.872 +/- 0.014) was significantly greater statistically (P <0.001) than that for LRA (0.791 +/- 0.006). The simplified ANNA models that used the pMRI findings in addition to PSA and Gleason score were as accurate as the model that used all the variables (P = 0.89). A high percentage of cancer in the biopsy specimens, pMRI findings, and high PSA density were equally the most influential predictors (relative weight 1.881, 1.964, and 1.493, respectively). CONCLUSIONS: All the ANNA models in this study were superior to LRA in the prediction of PSMs. The ANNA using pMRI findings, PSA level, and Gleason score as input variables performed as well as the ANNA using all the input parameters. Additional studies seem warranted. PMID- 15351583 TI - Prostate-specific antigen, sex steroid hormones, and the insulin-like growth factor axis in U.S.-born, Jamaican, and Haitian black men: a pilot study. AB - OBJECTIVES: African-Caribbean men have a risk of prostate cancer comparable to that of African-American men. To begin exploring potential risk factors for prostate cancer in these high-risk black subgroups, we conducted a pilot study in Brooklyn, New York, a community with large numbers of African-Americans and immigrants from Jamaica and Haiti. METHODS: Black men, 35 to 65 years of age, who were born in the United States, Jamaica, or Haiti were recruited in Brooklyn. The subjects' serum samples were analyzed for prostate-specific antigen (PSA) and the following hormones, which may be related to prostate cancer: testosterone, sex hormone-binding globulin, 3alpha-androstanediol glucuronide, insulin-like growth factor-1 (IGF-1), and IGF-binding protein-3 (IGFBP-3). Subgroup differences in PSA and hormonal levels, adjusted for relevant covariates, were explored using analysis of variance techniques. RESULTS: For 3 months, we recruited 21 U.S. born, 20 Jamaican-born, and 24 Haitian-born black men using various methods. The mean age-adjusted PSA level was 1.04 ng/mL in the U.S.-born men, 1.09 ng/mL in the Jamaican-born men, and 0.85 ng/mL in the Haitian-born men (P = 0.55). The mean age-adjusted hormone levels, as well as testosterone/sex hormone-binding globulin and IGF-1/IGFBP-3 ratios, also were not significantly different statistically across the subgroups. CONCLUSIONS: It is feasible to conduct epidemiologic studies of prostate cancer in these high-risk black subgroups in Brooklyn. Our preliminary data suggest that the serum levels of PSA and potential hormonal risk factors are similar among U.S.-born, Jamaican-born, and Haitian born black men. Larger follow-up studies are being planned to confirm these findings. PMID- 15351584 TI - Unilateral pudendal nerve blockade for relief of all pain during transrectal ultrasound-guided biopsy of the prostate: a randomized, double-blind, placebo controlled study. AB - OBJECTIVES: To investigate the efficacy of unilateral pudendal nerve block for the relief of all pain during transrectal ultrasound (TRUS)-guided prostate biopsy. TRUS-guided prostate biopsy is the standard procedure to diagnose or rule out prostate cancer. The pain, attributed to ultrasound probe insertion and the needle punctures into the prostate, inflicted by TRUS-guided prostate biopsy limits its effectiveness. METHODS: We performed a prospective, randomized, double blind, placebo-controlled study of 65 consecutive men suspected of having prostate cancer who were undergoing TRUS-guided prostate biopsy, 51 of whom fulfilled the inclusion criteria. Before the biopsy, each patient was randomized to one of two groups. Both the patient and the physician who performed the TRUS guided biopsy were unaware of the contents of the injection for the pudendal nerve block. Unilateral pudendal nerve blockade was performed transperineally with digital rectal examination guidance using 10 mg of 1% prilocaine (group 1 [n = 26]) or 10 mL of a 0.9 NaCl solution (group 2 [n = 25]) by way of a 22-gauge spinal needle by the same anesthetist. Pain was evaluated using an 11-point visual analog scale questionnaire. RESULTS: No statistically significant differences were found in the visual analog scale score for pain during the pudendal nerve blockade or digital rectal examination between the groups. A statistically significant difference was found in the visual analog scale score for the biopsy procedure (P < 0.01) and probe discomfort (P < 0.05) between the two groups. CONCLUSIONS: Unilateral pudendal nerve blockade was effective in reducing the pain at both biopsy and probe manipulation in our study. PMID- 15351585 TI - Influence of transrectal ultrasound probe on prostate cancer detection in transrectal ultrasound-guided sextant biopsy of prostate. AB - OBJECTIVES: To determine whether end-fire probes increase the prostate cancer (PCa) detection rate. Enhancing the PCa detection rate is the main goal of biopsy protocols. Prostate biopsy is limited by side-fire probes to a longitudinal axis, but end-fire probes allow biopsy cores to also be taken in the transverse section. METHODS: A total of 2625 patients underwent systematic sextant biopsy in three institutions using the same protocol. Three different ultrasound probes were used-the Kretz Combisone and Bruel & Kjaer side-fire probes and the ATL HDI end-fire probe. We retrospectively evaluated the influence of the probe on the PCa detection rate. RESULTS: The Kretz probe was used in 384 men, the Bruel & Kjaer probe in 598 men, and the ATL probe in 1643 men. Overall, 35.2% had PCa detected. Analyzing all patients, no statistically significant difference (P = 0.73) was found for the probes, but the subgroup with a prostate-specific antigen level of 4 to 10 ng/mL demonstrated a statistically significant improvement in the detection rate using the end-fire probe (31.3% versus 24.5% and 21.5% for the side-fire probes, P = 0.01). Patients with nonpalpable PCa also demonstrated a statistically significant increase in detection with the end-fire probe (P = 0.004). Multivariate analysis confirmed that the ultrasound probe is an independent parameter to enhance the PCa detection rate. CONCLUSIONS: Our results showed that end-fire probes provide a statistically significant improvement in the PCa detection rate compared with side-fire probes in patients with a prostate specific antigen level of 4 to 10 ng/mL and nonpalpable disease. The reason could be the facilitated sampling in the most lateral part of the peripheral zone. Our results suggest that the widespread use of end-fire probes for prostate biopsy could enhance the PCa detection rate. PMID- 15351586 TI - Effect of dutasteride on the detection of prostate cancer in men with benign prostatic hyperplasia. AB - OBJECTIVES: To examine the rate of prostate cancer detection in three large randomized placebo-controlled benign prostatic hyperplasia trials of dutasteride. Dutasteride, which lowers serum dihydrotestosterone more than 93% by inhibiting type 1 and type 2 5-alpha-reductase, is effective in the treatment of benign prostatic hyperplasia. However, its effect on the development of prostate cancer is unknown. METHODS: A total of 4325 men with benign prostatic hyperplasia but without a history, or evidence, of prostate cancer, and a serum prostate-specific antigen level of 1.5 to 10 ng/mL, were randomized to 0.5 mg/day dutasteride or placebo for 24 months. The prostate cancer detection rates for subjects were determined by non-protocol-mandated biopsies, either during the double-blind phase of the study or during the first 3 months of the open-label extension. A follow-up questionnaire was administered to a subset of consenting subjects to ascertain the number, outcomes, and reasons for the prostate biopsies. RESULTS: The cumulative incidence of prostate cancer as an adverse event was significantly lower in the dutasteride versus placebo group at 24 months (1.1% versus 1.9%, P = 0.025) and 27 months (1.2% versus 2.5%, P = 0.002). There were no differences in the diagnosis rates of prostate cancer during the first 15 months, after which time the detection rate of prostate cancer increased in the placebo group and remained low in the dutasteride group. CONCLUSIONS: Prostate cancer detection was significantly lower in subjects randomized to dutasteride compared with the placebo group. These results have prompted the initiation of the Reduction by Dutasteride of Prostate Cancer Events (REDUCE) study, which was designed and powered to test the hypothesis that treatment with dutasteride decreases the incidence and progression of prostate cancer. PMID- 15351590 TI - Relating biopsy and clinical variables to radical prostatectomy findings: can insignificant and advanced prostate cancer be predicted in a screening population? AB - OBJECTIVES: To assess the capacity of several clinical and needle biopsy pathologic parameters to predict insignificant and advanced prostate carcinoma (CaP) in radical prostatectomy tissue from men enrolled in a prostate-specific antigen screening program. METHODS: We captured multiple clinical variables and measures of needle biopsy tumor extent from 152 men with Stage T1c CaP with a mean of six biopsy cores who were treated with radical prostatectomy. Insignificant CaP was defined as a tumor volume of less than 0.5 cm(3) that was organ confined with a Gleason score less than 7. Advanced CaP was defined by a formula that combined the Gleason score, pathologic stage, and margin status. Bivariate and logistic regression analyses were used to identify variables predictive of either insignificant or advanced CaP. RESULTS: Of the cases of CaP, 25.7% were pathologically insignificant, and 14.5% were pathologically advanced. The best model for predicting insignificant CaP was less than 10% tumor as the greatest percentage of carcinoma in any core and a biopsy Gleason score of less than 7, yielding a sensitivity of 76.9% and specificity of 75.2%. For predicting advanced CaP, the best model was a total biopsy length of CaP greater than 3 mm, Gleason high-grade pattern 4 or 5 disease, perineural invasion in the biopsy, and more than one in six biopsy cores containing CaP, yielding a sensitivity of 13.6% and specificity of 100%. CONCLUSIONS: The prediction of insignificant and advanced CaP on an individual basis in patients from a prostate-specific antigen screening study is a challenging problem. However, several histopathologic features of CaP in needle biopsy tissue contain useful information about the severity of disease. PMID- 15351591 TI - Positive proximal (bladder neck) margin at radical prostatectomy confers greater risk of biochemical progression. AB - OBJECTIVES: To identify the prognostic significance of positive bladder neck margin as the sole positive margin after radical retropubic prostatectomy for treatment of prostate cancer. METHODS: We retrospectively analyzed the data from patients who underwent radical retropubic prostatectomy at Johns Hopkins Hospital between 1984 and 2001. Of 164 patients with positive margins, 38 had a positive margin only at the bladder neck margin and 126 had a positive margin at one or more locations separate from the bladder neck margin. RESULTS: Kaplan-Meier analyses showed that patients with a positive bladder neck margin had a greater biochemical progression risk than patients with positive margins at one or more locations other than the bladder neck margin. The 5-year actuarial risk of progression in men with positive and negative bladder neck margins was 69.8% and 33.0%, respectively. A positive bladder neck margin was statistically an independently significant predictor of progression (P = 0.001), along with the prostatectomy Gleason score (P = 0.0001) in the multivariate analysis. The hazard ratio for predicting progression for postoperative Gleason score and bladder neck positivity was 2.1 and 2.5, respectively. Serum prostate-specific antigen values, clinical stage, age, and the percentage of tumor at the bladder neck margin were not predictors of progression in either univariate or multivariate analysis. CONCLUSIONS: Patients with a positive bladder neck margin as the sole positive margin at radical retropubic prostatectomy have a greater risk of biochemical progression than patients with one or more positive margins at other locations, although not as adverse as the risk reported for those with Stage T4 disease. PMID- 15351592 TI - High-dose-rate iridium-192 afterloading therapy combined with external beam radiotherapy for T1c-T3bN0M0 prostate cancer. AB - OBJECTIVES: To determine the efficacy of radiotherapy (RT) for T1c-T3bN0M0 prostate cancer in a prospective clinical trial of concurrent external beam RT and fractionated iridium-192 high-dose-rate brachytherapy. METHODS: Included in the study were 98 patients with T1c-T3bN0M0 prostate cancer who were diagnosed between October 1997 and September 2002 and underwent high-dose-rate brachytherapy with external beam RT. Treatment consisted of external beam RT (four ports) to the prostate of 16 fractions of 2.3 Gy to a total dose of 36.8 Gy and high-dose-rate brachytherapy of 4 fractions of 6.0 Gy within 30 hours to a total dose of 24.0 Gy. No patient received adjuvant hormonal therapy after RT. RESULTS: The most recent prostate-specific antigen level was 0.0 to 3.9 ng/mL (median 0.4). Seven patients (7.1%) developed recurrence and treatment was considered a failure (bone metastasis in two and biochemical failure in five). The overall biochemical disease-free survival (bDFS) rate was 95.9% at 2 years and 92.9% at 5 years. The bDFS rate by T stage was 98.6% at 2 years and 95.9% at 5 years for Stage T1c-T2b and 88.0% and 84.0% for Stage T3a-b, respectively (P = 0.047). The 2-year and 5-year bDFS rate was better in patients with an initial prostate-specific antigen level of less than 20 ng/mL compared with 20 ng/mL or greater (98.6% and 97.1% versus 93.1% and 82.8%, respectively, P = 0.0261). Acute toxicity was mild to moderate (Radiation Therapy Oncology Group grade 1-2) and consisted of cystourethritis or proctitis in 29 (29.6%) of 98 patients. CONCLUSIONS: With a low complication rate and satisfactory bDFS rates, this combination therapy can be considered an alternative method for clinical Stage T1c-T3b prostate cancer and is expected to improve patient quality of life. Additional long-term follow-up is needed to confirm this treatment. PMID- 15351593 TI - Intraoperative characterization of arterial vasculature in spermatic cord. AB - OBJECTIVES: To characterize the arterial microanatomy within the spermatic cord. METHODS: Between June 2002 and March 2003, 72 men underwent 120 microsurgical subinguinal varicocelectomies for the treatment of varicocele. A micro-ruler was used to measure the diameter of each artery in the spermatic cord at the external inguinal ring. RESULTS: A single testicular artery was identified in 83 (69%), two testicular arteries in 32 (27%), and three arteries in 4 (3%) of 120 cases. The testicular artery diameter ranged from 0.2 to 1.9 mm (mean 1.0). A single vasal artery was identified in 117 (98%) of 120 cases, with the vasal artery diameter ranging between 0.2 and 1.8 mm (mean 0.6). A single cremasteric artery was identified in 66 (55%), two arteries in 37 (31%), and three arteries in 5 (4%) of 120 cases. The cremasteric artery diameter ranged from 0.1 to 1.5 mm (mean 0.5). The diameter of the testicular artery was equal to or greater than the sum of the vasal and cremasteric artery diameters in 69 (57.5%) of 120 spermatic cords. CONCLUSIONS: The testicular artery was consistently the largest caliber arterial vessel, with a diameter equal to or greater than the sum of the vasal and cremasteric arteries in more than 50% of the spermatic cords. This suggests that although the vasal and cremasteric arteries combined contribute significantly to the testicular blood supply, the testicular artery provides most of the blood flow to the human testes. Preservation of the testicular arteries is recommended for optimal testicular blood flow. PMID- 15351594 TI - Heroic measures may not always be justified in extensive urethral stricture due to lichen sclerosus (balanitis xerotica obliterans). AB - OBJECTIVES: Strictures due to lichen sclerosus (LS) may affect the urethra as far proximally as the mid-bulb. For such strictures, a staged full-length repair is required and should use a nonpenile graft source such as buccal mucosa. Many cases occur in a population already accustomed to seated voiding, leading us to re-evaluate this approach and, in some circumstances, recommend definitive perineal urethrostomy alone. METHODS: We reviewed the medical records and retrograde urethrograms of all patients undergoing surgery for LS at our facilities between January 1991 and June 2002. RESULTS: A total of 63 patients, with an average age of 54.2 years, underwent surgery for LS stricture with an average follow-up of 38.5 months (range 4 to 117). Of the 63 patients, 19 underwent grafting in preparation for future reconstruction. Of these, 11 completed the second-stage repair, and 8 patients elected not to undergo the second stage of the repair, leaving a functional perineal urethrostomy. This led us to look more critically at definitive perineal urethrostomy alone for some patients. Parallel with the staged repairs, and subsequent to them, 44 patients underwent perineal urethrostomy alone. CONCLUSIONS: The often extensive nature of LS, the prevailing philosophy that urethroplasty must use nonpenile skin, the limited availability of such sources, and the acceptance of many patients for seated voiding makes definitive perineal urethrostomy alone a viable treatment option. In all our cases, this satisfied patients' quality of life concerns, leaving the anterior urethra dry and amenable to future repair. Younger men desirous of penile voiding should still be considered for staged repair using current techniques. PMID- 15351595 TI - Encrusted cystitis and pyelitis in children: an unusual condition with potentially severe consequences. AB - OBJECTIVES: To report our experience with the management of encrusted cystitis and pyelitis (EC and EP) in the pediatric population. EC and EP are well-known entities in adults but are rarely identified in children. They consist of mucosal encrustations and are due to specific microorganisms. METHODS: Between 1996 and 2001, 4 children with a mean age of 9 years (range 4 to 13) were treated for EC (n = 2), EP (n = 1), and EC and EP (n = 1). The latter was a kidney transplant recipient. We retrospectively evaluated the clinical characteristics of the patients and the results of conservative management. RESULTS: The delay between the beginning of the symptoms and the diagnosis was longer than 1 month in all cases. The diagnosis of EC was not evoked and was made during cystoscopy in all cases. EP was diagnosed during pyelotomy in 1 patient because it was evoked and confirmed by computed tomography scan in the kidney transplant recipient. Corynebacterium urealyticum was identified in the urine of all patients. EC was treated by antibiotics and endoscopic debulking, and EP was treated by antibiotics and local acidification. The duration of antibiotic therapy was between 1 and 6 months. The tolerance to local acidification of the kidneys was poor. Cure was achieved in 3 cases, but the treatment of EP failed in the kidney transplant recipient and graft removal was decided after 6 months of failed management because intractable febrile urinary tract infections became life threatening for the patient. CONCLUSIONS: EC and EP are uncommon in children; however, these diseases must be considered. They must be diagnosed rapidly and require, if possible, conservative management. Nevertheless, kidney loss can occur in transplant recipients with EP. PMID- 15351596 TI - High-grade renal injuries in children--is conservative management possible? AB - OBJECTIVES: To review our experience with the management of high-grade (grade IV and V) renal injuries to clarify the role of conservative management. METHODS: From 1991 to 2003, 79 consecutive patients (age range 2 to 14 years) with renal injuries were treated in an urban level I pediatric trauma center. Twenty children were identified as having high-grade renal injury (grade IV, 10 children and grade V, 10 children). The mechanism of injury was blunt trauma in 17 patients (85%) and penetrating trauma in 3 (15%). RESULTS: Of the 10 patients with grade IV injury, 8 (80%) were successfully treated conservatively with bedrest and catheter drainage. Two patients with persistent urine leaks required ureteral stenting, and one subsequently required open operative repair. The initial radiographic findings in both patients demonstrated complete renal fracture with retained vasculature to both renal segments. All 10 patients with grade V injury required open operative management and only 3 (30%) achieved long term renal salvage. CONCLUSIONS: Most children with grade IV renal injury can be treated conservatively. Patients with complete renal fracture or significant urinary extravasation on initial radiographic imaging may be less likely to undergo spontaneous resolution. Patients with a persistent urinary leak can be successfully treated with internal drainage. Grade V injuries are associated with an increased risk of requiring open operative intervention, and the renal preservation rates are low. PMID- 15351597 TI - Abdominal sacral colpopexy for repair of neovaginal prolapse in male-to-female transsexuals. AB - Penectomy, bilateral orchiectomy, and penoscrotal flap vaginoplasty are procedures that increase the psychosocial well-being and enhance body acceptance of male-to-female transsexuals. The incidence of neovaginal prolapse is not known but is believed to be relatively rare. We report 2 cases of neovaginal prolapse that were successfully treated with abdominal sacral colpopexy at our institution. PMID- 15351598 TI - Extraction of UroLume endoprostheses with one-stage urethral reconstruction. AB - We describe a technique for transperineal excision of UroLume endoprostheses with one-stage urethral reconstruction. We have used this technique in 3 patients with occluded urethral stents placed for bulbar stricture disease. All patients experienced acceptable outcomes. PMID- 15351599 TI - Heterotopic ossification in testicular microlithiasis. PMID- 15351600 TI - Fournier's gangrene as a complication of steroid enema use for treatment of radiation proctitis. PMID- 15351601 TI - Rapidly growing adenomatoid tumor extending into testicular parenchyma mimics testicular carcinoma. AB - A 40-year-old man presented with a 1-month history of a large, painless, rapidly growing right scrotal mass. Examination, ultrasonography, and intraoperative inspection did not exclude an intratesticular tumor, and a radical orchiectomy was performed. Histology was typical for adenomatoid tumor, but gross and microscopic inspection revealed bundles of tumor extending from the tunica albuginea into the testicular parenchyma. In this case the rapid growth, large size, and the inability to exclude an intratesticular origin directed suspicion toward a malignant neoplasm. In hindsight, the tumor's unique pattern of extension into the adjacent testicular parenchyma confounded the preoperative diagnosis. PMID- 15351602 TI - Furosemide nephrolithiasis causing ureteral obstruction and urinoma in a preterm neonate. AB - Nephrolithiasis and nephrocalcinosis in low-birth-weight, preterm neonates are well-documented complications of furosemide administration. Perinephric urinomas and urinary ascites have been previously reported as complications of obstruction due to posterior urethral valves or ureteropelvic junction obstruction. We present an unusual case of furosemide-induced nephrolithiasis causing ureteral obstruction, urinoma, and acute renal failure in a preterm neonate. PMID- 15351603 TI - Intracranial metastatic prostate carcinoma presenting as intermittent double vision. AB - We describe a prostate cancer patient whose initial symptom was intermittent double vision. Intracranial magnetic resonance imaging demonstrated a pituitary mass extending to the cavernous sinus, which caused the double vision. After hormonal therapy for prostate cancer, the pituitary mass disappeared, and double vision was completely resolved without local therapy for the brain. In the 19 months of follow-up after hormone treatment, the prostate cancer remained stable, and the patient remained neurologically intact. PMID- 15351604 TI - Use of a mechanical thrombectomy catheter for percutaneous extraction of renal fungal bezoars in a premature infant. AB - Fungal urinary tract infections are commonly encountered in the hospitalized neonate. Although these infections most commonly take the form of cystitis, the infection may be complicated by the formation of fungal bezoars, with subsequent urinary tract obstruction. In certain cases, endosurgical debulking or extraction of the fungal bezoar may be necessary. This is particularly challenging in neonates due to their often-compromised physiologic state and small size. We report a case of a premature infant with bilateral obstructing renal fungal bezoars in whom a percutaneous catheter-based thrombectomy system was used successfully to debulk the fungal burden. PMID- 15351605 TI - Incomplete bladder duplication presenting antenatally. AB - We report a case of an incomplete bladder duplication diagnosed by prenatal ultrasonography at 35 weeks of gestation. The patient was followed until bladder neck obstruction and high-grade vesicoureteral reflux of a solitary kidney prompted definitive repair at 6 months of age. Bladder duplication is a rare anomaly. To our knowledge this is the first documented case of an incomplete bladder duplication discovered antenatally. PMID- 15351606 TI - Monoamniotic twin discordant for bilateral renal agenesis with normal pulmonary function. AB - A 22-year-old G4P3 woman with a twin pregnancy underwent an uncomplicated delivery at 35 weeks of gestation of normal-appearing twin boys. One of the boys returned 24 hours later with no voiding since birth. He was doing well with no respiratory difficulties and normal physical examination findings. Renal/bladder ultrasonography and computed tomography of the abdomen and pelvis revealed no evidence of kidneys. The admission laboratory values were consistent with renal failure. A peritoneal dialysis catheter was inserted and peritoneal dialysis started. Our patient died at 2 months old secondary to complications from the peritoneal dialysis. We describe the unusual case of a monoamniotic twin discordant for bilateral renal agenesis with normal pulmonary function. This case shows the importance of normal renal function in the production of amniotic fluid and its relationship to the proper development of the pulmonary system. PMID- 15351607 TI - Malignant B-cell lymphoma in renal cyst wall. AB - Renal cysts are frequently found in adults older than 50 years of age. Bosniak type III and IV cysts are commonly associated with malignancy, but most Bosniak I and II lesions are benign, and the optimal management has not been clearly defined. Although computed tomography and ultrasound examinations have improved diagnostic accuracy, some masses will remain indeterminate and require more invasive evaluation. We report a patient with a Bosniak type II renal cyst associated with malignant B-cell lymphoma in the cyst wall diagnosed after laparoscopic renal exploration. PMID- 15351608 TI - Intrascrotal extratesticular arteriovenous malformation. AB - We describe a case of intrascrotal-extratesticular arteriovenous malformation in an adult that presented as a hard scrotal mass mimicking post-traumatic hematoma. This is the first reported case of an arteriovenous malformation at the intrascrotal-extratesticular region. PMID- 15351609 TI - Urinary retention after tension-free vaginal tape procedure: from incision to excision...to complete urethrolysis. AB - We report on a healthy neurologically intact 45-year-old patient with persistent obstruction after a tension-free vaginal tape procedure necessitating several surgical procedures for correction. PMID- 15351610 TI - Bilateral spontaneous perinephric urinomas: case report and review of the literature. AB - Urinoma is defined as an encapsulated collection of extravasated urine in the perirenal or paraureteral space. Urinomas in childhood are rare and usually either secondary to obstructive uropathies or due to blunt or penetrating trauma leading to injuries of the collecting system and/or ureter. Spontaneous perinephric urinomas are rare. We report the case of an 8-year-old boy with bilateral spontaneous perinephric urinomas secondary to an abnormally elevated voiding pressure with detrusor sphincter dyssynergia. The available literature on this subject is reviewed, and clinical features, pathophysiology, and treatment of this entity are discussed. PMID- 15351611 TI - Histopathologic confirmation of complete cancer-cell kill in excised specimens after renal cryotherapy. AB - We present the first case of complete documentation of total cancer ablation in a previously cryoablated human renal tumor. A 61-year-old patient with metachronous bilateral renal cell carcinoma was treated with open partial nephrectomy and subsequently laparoscopic cryoablation. His renal function deteriorated, prompting bilateral radical nephrectomy before renal transplantation. Detailed histopathologic examination of the specimen did not reveal any evidence of malignancy in two cryoablated sites 36 months (left kidney) and 19 months (right kidney) after the respective cryoablations. PMID- 15351612 TI - Misattributed paternity in a living related donor: to disclose or not to disclose? AB - Many ethical considerations surround living kidney donation, some of which are not anticipated. We present a case in which misattributed paternity was inadvertently discovered during the workup of a father and son and present arguments for and against disclosure of this information. We recommend that transplant programs advise patients participating in living organ donor programs that misattributed paternity might be discovered during routine preoperative testing and that protocols for dealing with complex ethical issues be in place. PMID- 15351613 TI - Successful treatment of primary bilateral renal mucormycosis with bilateral nephrectomy. AB - Bilateral invasive renal mucormycosis has previously been associated with a 100% mortality rate. We report a case of bilateral invasive renal mucormycosis in a patient treated with amphotericin B and bilateral nephrectomy who survived and is currently disease-free. PMID- 15351614 TI - Penile strangulation treated with the modified string method. AB - Penile strangulation by a nonmetallic or thin metallic ring is easily overcome by severing the object, but a heavy metal ring causing penile strangulation is very difficult to sever. Here we report 2 cases of penile strangulation by metal rings, involving a 38-year-old man and a 44-year-old man, both of whom were treated with the modified string method and glandular puncture. PMID- 15351615 TI - Laparoscopic warm renal ischemia in the solitary porcine kidney model. AB - OBJECTIVES: To determine the effect of a laparoscopic approach on warm renal ischemia in the pig with a solitary kidney. Although the maximal safe duration for warm ischemia during open partial nephrectomy is commonly accepted to be 30 minutes, the maximal safe ischemic time during laparoscopic partial nephrectomy has not been previously determined. METHODS: Sixteen farm pigs underwent unilateral laparoscopic right nephrectomy. Two weeks later, the pigs underwent complete laparoscopic mobilization of the remaining left kidney and were randomized to complete hilar clamp times of 0, 30, 60, or 90 minutes. Serum creatinine was evaluated before right nephrectomy and at days 0, 2, 4, 7, 14, and 30 after renal ischemia during laparoscopy. All renal specimens were evaluated by an experienced nephropathologist in a blinded fashion. RESULTS: The serum creatinine remained stable at all points in the control and 30-minute ischemia groups. The serum creatinine level rose initially on days 2 and 4 in the 60 minute and 90-minute ischemia groups but returned to baseline by day 7. At harvest, no statistically significant difference was found among the groups in serum creatinine or histologic features. CONCLUSIONS: Renal ischemic times up to 90 minutes during laparoscopy were well tolerated by the solitary porcine kidney. Possible explanations for this finding include the protective effect of a solitary kidney and the potential protective effect of relative ischemic preconditioning provided by the pneumoperitoneum. PMID- 15351616 TI - Cytotoxicity of different intracavernous vasoactive drugs on cultured endothelial cells of human corpus cavernosum penis. AB - OBJECTIVES: To investigate the cytotoxic effect of prostaglandin E(1) (PGE(1)), a standard combination of papaverine/phentolamine, and a triple mixture of these agents on human cavernosal endothelial cells using a cell culture model. The endothelial layer of the corpus cavernosum plays an important role in signal transduction of penile erection and is directly exposed to vasoactive agents after intracavernous injection for erectile dysfunction. METHODS: Primary endothelial cells were obtained from the corpus cavernosum of 13 potent patients undergoing penile surgery. Cultured cells were exposed for 30 minutes to physiologic dilutions of 20 microg PGE(1), 30 mg papaverine/1 mg phentolamine, or the same dosages of the triple mixture of these agents, each dissolved in 5 to 50 mL sodium chloride. Lactate dehydrogenase release as a cytotoxicity marker was measured 6 hours after drug exposure, and the total cell metabolic activity was quantified after 48 hours with a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS)-based assay. Additionally, the amount of viable cells was identified with a dual fluorescent staining procedure. RESULTS: The initial release of lactate dehydrogenase was elevated up to 3.2-fold in the concentrated papaverine/phentolamine and triple mixture group compared with PGE(1) and the control. After 48 hours, the papaverine-containing formulations led to a significant dose-dependent decrease in the viable cell count and metabolic activity of the cultures that was not noticed with PGE(1). CONCLUSIONS: These in vitro data strongly suggest an unfavorable effect of vasoactive agents containing papaverine on cavernosal endothelial cells. Before fibrotic changes of the smooth muscle stroma, the functionally important endothelium of the corpus cavernosum might suffer significantly from intracavernous injection therapy. Therefore, papaverine should no longer be used for this indication. PMID- 15351617 TI - Role of adenosine triphosphate-dependent potassium channels in canine penile erection. AB - OBJECTIVES: To define the physiologic role and hemodynamic features of nitric oxide (NO) and the adenosine triphosphate (ATP)-dependent K(+) (K(ATP)) channel in canine penile erection. METHODS: Mongrel dogs were anesthetized, and penile erection was induced by electrical stimulation of the pelvic nerve. Changes in the intracavernous pressure (ICP) were measured with a transducer. RESULTS: The basal ICP was 12.8 +/- 5.0 mm Hg. Pelvic nerve stimulation (5 to 20 V, 5 to 15 Hz, for 1-minute intervals) significantly increased the ICP to 86.2 +/- 11.4 mm Hg (n = 5, P <0.05). Treatment with the NO synthase inhibitor N(omega)-nitro-L arginine methyl ester (10 mg/kg intravenously) abolished this increase (15.4 +/- 5.0 mm Hg, n = 5). Intracavernous injection of the K(ATP) channel opener cromakalim (3 and 10 microg) increased the ICP (103 +/- 14.4 mm Hg and 106 +/- 12.1 mm Hg, respectively; n = 4). This response was abolished by the prior intracavernous injection of the selective K(ATP) channel-specific blocker glibenclamide (10 mg). Glibenclamide did not affect the increase in ICP induced by electric stimulation of the pelvic nerve (88 +/- 24.2 mm Hg). CONCLUSIONS: Our results indicate that relaxation of canine cavernous smooth muscle and penile tumescence are mediated by NO. The failure of glibenclamide to affect the increase in ICP induced by pelvic nerve stimulation suggests that ATP-dependent K(+) channels probably do not play a physiologic role in canine penile erection. PMID- 15351618 TI - Effect of timing of diuresis on response of rat bladder to outlet obstruction. AB - OBJECTIVES: To determine whether results similar to a previous study, in which sucrose-induced diuresis started before creating partial outlet obstruction resulted in protection of bladder contractile function in obstructed animals, would be obtained if diuresis were begun after creation of partial outlet obstruction. METHODS: Thirty-six rats were separated into six groups of 6 rats each: group 1, control; group 2, diuresis; group 3, obstructed; group 4, obstructed immediately after diuresis; group 5, obstructed with diuresis beginning 10 days after obstruction; and group 6, obstructed before diuresis. Diuresis was induced by feeding 5% sucrose instead of water. After 3 weeks of obstruction, all bladders were excised, cut into longitudinal strips, and mounted in isolated muscle baths for physiologic study. RESULTS: Sucrose-induced diuresis resulted in a statistically significant increase in bladder mass and a mild increase in smooth muscle contractility compared with the control water-fed group. Obstruction resulted in a fourfold increase in bladder weight and substantial decreases in the responses to all forms of stimulation. The bladder weights of the obstruction plus diuresed groups were all similar to the diuresis alone group and significantly lower than those of the obstructed group. All diuresed groups showed protection of the contractile responses compared with the obstruction-alone group. Unexpectedly, the group that began diuresis 1 day after obstruction showed the strongest protection. CONCLUSIONS: Diuresis significantly reduced the increase in bladder mass induced by obstruction and also reduced the level of contractile dysfunction. Diuresis begun 1 day after obstruction resulted in the greatest level of protection. PMID- 15351619 TI - Expression of p27 and its ubiquitin ligase subunit Skp2 in upper urinary tract transitional cell carcinoma. AB - OBJECTIVES: To analyze p27 and S-phase kinase-associated protein 2 (Skp2) expression in upper urinary tract transitional cell carcinoma (TCC) with respect to biologic significance. p27 (p27/kip1) is involved in cell cycle control, and loss of p27 protein expression may result in tumor development and/or progression. The association of p27 with the ubiquitin ligase subunit Skp2 targets p27 for degradation. METHODS: A total of 53 upper urinary tract TCC specimens were investigated immunohistochemically using a tissue microarray technique. The immunoreactivity of p27 and Skp2 was analyzed with respect to associations with pT stage, grade, and prognosis. RESULTS: Non-neoplastic renal tissue showed p27 immunoreactivity in tubule epithelium and pelvic urothelium, but lacked immunoreactivity for Skp2. In the TCC specimens, p27 immunoreactivity was noted in 47 (89%) of 53 cases. High p27 expression (50% or greater of tumor cell nuclei) tended to decrease with rising tumor stage (14 [45%] of 31 with pT1 pT2 versus 4 [18%] of 22 with pT3; P = 0.076), but was independent of tumor grade (11 [39%] of 28 grade 2 versus 7 [28%] of 25 grade 3-4; P = 0.56). Skp2 immunoreactivity was noted in 32 (60%) of 53 tumors. Skp2 expression increased with rising tumor stage (9 [41%] of 22 pT1 versus 23 [74%] of 31 pT2-pT3; P = 0.023) and tumor grade (12 [43%] of 28 grade 2 versus 20 [80%] of 25 grade 3; P = 0.043) and was associated with angioinvasion (P = 0.017). In multivariate analysis, tumor stage proved to be the only independent prognostic factor regarding disease-free survival. CONCLUSIONS: p27 and Skp2 are additional biomarkers in urogenital pathologic findings. The statistically significant association of Skp2 expression with high-grade TCC, as well as the lack of expression in non-neoplastic tissue, suggests that Skp2 could be a promising target for future cancer therapy strategies. PMID- 15351620 TI - Numeric aberrations of HER-2 and chromosome 17 detected by fluorescence in situ hybridization in urine-exfoliated cells from patients with urothelial carcinoma. AB - OBJECTIVES: To elucidate the clinical significance of the HER-2 gene alterations in urine-exfoliated cells detected by fluorescence in situ hybridization (FISH) in patients with urothelial transitional cell carcinoma. METHODS: The relative increase of HER-2 (RI-HER2) and gain of chromosome 17 (G-17) in urine-exfoliated cells were examined using DNA probes for HER-2 and the chromosome 17 centromere in 103 patients. In addition, FISH analysis was performed using corresponding paraffin-embedded tissue sections from 45 cases to compare the results obtained using urine-exfoliated cells and those obtained using paraffin-embedded tissue. RESULTS: RI-HER2 and G-17 was found in 23 (22.3%) and 46 (44.6%) of 103 patients, respectively. RI-HER2 was significantly more frequent in tumors with two or more recurrences (40.7% versus 15.8%, P = 0.010) and in those with carcinoma in situ (CIS) (35.4% versus 15.9%, P = 0.029). G-17 was more frequent in high-grade tumors (69.1% versus 16.7%, P = 0.032), invasive tumors (63.6% versus 14.3%, P < 0.001), and in patients with CIS (77.1% versus 29.0%, P < 0.001). The positive rate for FISH (presence of RI-HER2 and/or presence of G-17) tended to be more frequent in FISH than in cytology. A comparison of the analyses using urine exfoliated cells and paraffin-embedded tissue showed identical results in 36 (80.0%) of 45 cases. CONCLUSIONS: Numeric alterations of the chromosome 17 centromere in urine-exfoliated cells detected by FISH may reflect the malignant potential of urothelial carcinoma. In addition, a relative increase in HER-2 was associated with the number of recurrences and the presence of CIS. PMID- 15351621 TI - Effects of C-peptide on expression of eNOS and iNOS in human cavernosal smooth muscle cells. AB - OBJECTIVES: To investigate the role of C-peptide alone or in conjunction with insulin on the expression of nitric oxide synthase (NOS) in human corpus cavernosum smooth muscle cells (HCSMCs). Erectile dysfunction, among diabetic patients, is a significant health problem. The specific causes of erectile dysfunction are unknown. It has been suggested that impairment of penile relaxation is related to a reduction of penile NOS. Plasma levels of C-peptide and insulin are decreased in individuals with type 1 diabetes and late-stage type 2 diabetes. METHODS: Primary cultures were initiated from explants of HCSMCs. Confluent cells at passages 2 to 4 were assigned to one of four groups with the following incubation conditions: (a) 27 mM glucose, (b) 27 mM glucose and insulin, (c) 27 mM glucose and human recombinant (hr)C-peptide, and (d) 27 mM glucose, insulin, and hrC-peptide. After 24 hours, total RNA and protein were extracted from cells and subjected to reverse transcriptase-polymerase chain reaction and Western blot analysis, respectively. Intracellular Ca(2+) was examined under the four conditions, using the Fura 2 method. RESULTS: The least expression of endothelial NOS (eNOS) and inducible NOS (iNOS) in HCSMCs was observed in cells exposed to 27 mM glucose alone. Increased expression of eNOS and iNOS was found after treatment with insulin or hrC-peptide alone, and the maximal expression of eNOS and iNOS was detected in HCSMCs exposed to both insulin and hrC-peptide. Western blot analyses using eNOS and iNOS antibodies confirmed the RNA data. These effects are likely mediated by the insulin-induced and/or C-peptide-induced increase in intracellular Ca(2+). CONCLUSIONS: Our results demonstrated that C-peptide, in the presence of insulin, increases the expression of iNOS and eNOS in HCSMCs. These results suggest that C-peptide, especially in conjunction with insulin, may have beneficial effects on cavernosal smooth muscle relaxation. PMID- 15351622 TI - Hugh Hampton Young, the anesthesiologist. PMID- 15351623 TI - Dissociable neural pathways for the perception and recognition of expressive and instrumental gestures. AB - Previous functional imaging studies have sought to characterize the neural correlates of gesture representation. However, little is yet known about the representation of different categories of gesture. Here we contrasted the perception of hand gestures that express inner feeling states, e.g. I am angry, I do not care, with the perception of instrumental gestures intended to change the behavior of others by communicating commands, e.g. come here, look over there. We hypothesised that recognition of expressive gestures would activate a network of brain regions associated with mentalising ('theory of mind') whereas instrumental gestures would activate different neural pathways. Twelve normal volunteers underwent fMRI while they watched a series of short videos (3 s duration) of actors performing expressive and instrumental gestures. The volunteers had either to recognise the gesture or to monitor the positions of the hands. As predicted, different neural networks were activated by the observation of instrumental or expressive gestures. The perception of expressive gestures elicited activity in the anterior paracingulate cortex, the amygdala and the temporal poles bilaterally and the right superior temporal sulcus. These regions have all previously been activated during the performance of mentalising tasks. In contrast, instrumental gestures elicited activity in a left-lateralised system previously associated with language and motor imitation. PMID- 15351624 TI - The space of senses: impaired crossmodal interactions in a patient with Balint syndrome after bilateral parietal damage. AB - Balint syndrome after bilateral parietal damage involves a severe disturbance of space representation including impaired oculomotor behaviour, optic ataxia, and simultanagnosia. Binding of object features into a unique spatial representation can also be impaired. We report a patient with bilateral parietal lesions and Balint syndrome, showing severe spatial deficits in several visual tasks predominantly affecting the left hemispace. In particular, we tested whether a loss of spatial representation would affect crossmodal interactions between simultaneous visual and tactile events occurring at the same versus different locations. A tactile discrimination task, where spatially congruent or incongruent visual cues were delivered near the patient's hands, was used. Following stimulation of the left hand in the left side of space, we observed visuo-tactile interactions that were not modulated by spatially congruent conditions. In contrast, performance following stimulation of the right hand in the right side of space was affected in a spatially selective manner--facilitated for congruent stimuli and slowed for incongruent stimuli. To dissociate effects on somatotopic and spatiotopic coordinates, we crossed the patient's hands during unimodal tactile discriminations. Tactile performance of the left hand improved when it was positioned in the right hemispace, whereas placing the right hand in left space produced no significant changes, suggesting that left-sided tactile inputs are coded with respect to a combination of limb- and trunk-centred coordinates. These data converge with recent findings in animals and healthy humans to indicate a critical role of the posterior parietal cortex in multimodal spatial integration, and in the fusion of different coordinates into a unified representation of space. PMID- 15351625 TI - Migration and fusion of tactile sensation--premorbid susceptibility to allochiria, neglect and extinction? AB - Migration of tactile sensation was found to occur very frequently in about 25% of normal people (High Error subjects) and very infrequently in others. When synchronous stimuli touched the two hands, if the unattended stimulus was modulated when the attended one was not (and was thus more salient) it "migrated" to the attended hand and fused with or replaced the attended stimulus. However, latencies reflecting congruence and incongruence of simultaneous stimuli showed that their identities on each hand had been (nonconsciously) registered veridically. Subsequent experiments, involving Focused and Divided Attention without speed pressure showed that mislocation errors in these subjects (i) were not due to confusion about location of otherwise perceptually distinct stimuli, (ii) nor to speed demand, (iii) nor to relative salience per se, (iv) were immune to attentional manipulation and practice in most subjects, (v) required a stimulus on the attended hand, and (vi) reflected a changed experience. Finally the same subjects rated similarity of the attended stimulus when accompanied by a distractor to each stimulus alone. Scaling distributions tested against a sampling model showed that most High Error subjects experienced a fusion of the stimuli, some experienced a replacement, and Low Error subjects experienced neither. The individual difference appears to be in attentional separability and spatial binding of tactile stimuli. Mislocation to the focus of spatial attention is common in healthy people, especially when binding is prevented. The present phenomenon appears equivalent to allochiria, but also accounts for phenomena in neglect and extinction, and suggests a premorbid susceptibility to spatial migration and integration that can be exaggerated by brain damage. PMID- 15351626 TI - The pathophysiology of letter-by-letter reading. AB - Pure alexia is a frequent and incapacitating consequence of left occipitotemporal lesions. It is thought to result from the disruption or the disconnection of the visual word form area (VWFA), a region reproducibly located within the left occipito-temporal sulcus, and encoding the abstract identity of strings of visual letters. Alexic patients often retain effective single letter recognition abilities, and develop an effortful letter-by-letter reading strategy which is the basis of most rehabilitation techniques. We study a patient who developed letter-by-letter reading following the surgical removal of left occipito-temporal regions. Using anatomical and functional MRI in the patient and in normal controls, we show that alexia resulted from the deafferentation of left fusiform cortex, and we analyze the network of brain regions subtending letter-by-letter reading. We propose that during letter-by-letter reading (1) letters are identified in the intact right-hemispheric visual system, with a central role for the region symetrical to the VWFA; (2) letters are serially transferred to the left hemisphere through the intact segment of the corpus callosum; (3) word identity is eventually recovered in the left hemisphere through verbal working memory processes involving inferior frontal and supramarginal cortex. PMID- 15351627 TI - Shared and distinct neurophysiological components of the digits forward and backward tasks as revealed by functional neuroimaging. AB - The digits forward (DF) and backward (DB) tasks are widely used neuropsychological measures believed to tap overlapping systems of phonological processing and working memory. Studies of focal brain lesions have partially elucidated the brain regions essential for these tasks; however relatively little information exists on the underlying functional neuroanatomy in the intact brain. We therefore examined the shared and separate neural systems of these tasks in two positron emission tomography (PET) experiments. In Experiment 1, eight healthy participants performed verbal DF, DB, and a sensorimotor control task during measurement of regional cerebral blood flow (rCBF). DF and DB each activated frontal, parietal, and cerebellar regions as well as prominently activating medial occipital cortex. To eliminate possible visuospatial confounds, Experiment 2 replicated the first experiment in six additional healthy participants who were blindfolded during the study. No differences in activation were found between the two experimental groups. Combined data from both experiments demonstrate that DF and DB rely upon a largely overlapping functional neural system associated with working memory, most notably right dorsolateral prefrontal cortex (DLPFC) and bilateral inferior parietal lobule (IPL) as well as the anterior cingulate, a region associated with attentional effort. The degree of activation increased linearly with increasing task difficulty in DF. DB additionally recruited bilateral DLPFC, left IPL, and Broca's area. Medial occipital cortex (including higher and lower visual processing areas) was robustly activated in both DF and DB and could not be attributed to visual processing per se, suggesting a possible visual imagery strategy for these aural verbal tasks. PMID- 15351628 TI - Action comprehension in aphasia: linguistic and non-linguistic deficits and their lesion correlates. AB - We tested aphasic patients' comprehension of actions to examine processing deficits in the linguistic and non-linguistic domains and their lesion correlates. Twenty-nine left-hemisphere injured patients and 18 age-matched control subjects matched pictured actions (with the objects missing) or their linguistic equivalents (printed sentences with the object missing) to one of two visually-presented pictures of objects. Aphasic patients performed poorly not only in the linguistic domain but also in the non-linguistic domain. A subset of the patients, largely consisting of severe and non-fluent aphasics, showed a greater deficit in the linguistic domain compared with the non-linguistic domain and across the patient group, deficits in the linguistic and non-linguistic domains were not tightly correlated. Poor performance in pantomime interpretation was associated with lesions in the inferior frontal, premotor and motor cortex, a portion of somatosensory cortex, and the caudate, while poor reading comprehension of actions was associated with lesions around the anterior superior temporal lobe, the anterior insula and the anterior portion of the inferior parietal lobe. Lesion size did not correlate with deficits. The lesion results for pantomime interpretation deficits demonstrate that lesions in the frontal component of the human analog of the "mirror neuron system" are associated with deficits in non-linguistic action understanding. For reading comprehension deficits, the lesion correlates are brain areas known to be involved in linguistic tasks including sentence processing and speech articulation; the parietal lesion site may also correspond to a subpart of the human mirror neuron system. These results indicate that brain areas important for the production of language and action are also recruited in their comprehension. Similar findings have been reported in electrophysiological and neuroimaging studies. Our findings now also lend neuropsychological support to an embodied view of brain organization for action processing. PMID- 15351629 TI - On the role of response conflicts and stimulus position for hemispheric differences in global/local processing: an ERP study. AB - It is widely assumed that the local and global levels of hierarchical stimuli are processed more efficiently in the left and right cerebral hemispheres, respectively. However, corresponding effects were not observed under all circumstances. In ERP studies, they occurred more often with centrally presented stimuli than with laterally presented ones, whereas reaction-time studies revealed that a response conflict between the levels is relevant. The present study examines which of these two factors is more important by presenting conflicting and non-conflicting stimuli to the left or right visual field and recording ERPs as well as collecting behavioral data. If a central stimulus position is crucial, then no effects should show up. Contrary to this prediction, the expected hemispheric differences were observed in the behavioral data as well as in the later occurring (N2 and P3) ERP amplitudes. However, in all variables, the effects were more pronounced for conflicting stimuli. The results suggest that response conflicts are more important for obtaining hemispheric differences in global/local processing than a central stimulus presentation. This is interpreted in the way that hemispheric differences vary with respect to the stimulus representation that is needed to select a proper response. PMID- 15351630 TI - Time course and hemispheric lateralization effects of complex pitch processing: evoked magnetic fields in response to rippled noise stimuli. AB - To delineate the time course and processing stages of pitch encoding at the level of the supratemporal plane, the present study recorded evoked magnetic fields in response to rippled noise (RN) stimuli. RN largely masks simple tonotopic representations and addresses pitch processing within the temporal domain (periodicity encoding). Four dichotic stimulus types (111 or 133 Hz RN at one ear, white noise to the other one) were applied in randomized order during either visual distraction or selective auditory attention. Strictly periodic signals, noise-like events, and mixtures of both signals served as control conditions. (1) Attention-dependent ear x hemisphere interactions were observed within the time domain of the M50 field, indicating early streaming of auditory information. (2) M100 responses to strictly periodic stimuli were found lateralized to the right hemisphere. Furthermore, the higher-pitched stimuli yielded enhanced activation as compared to the lower-pitch signals (pitch scaling), conceivably reflecting sensory memory operations. (3) Besides right-hemisphere pitch scaling, the relatively late M100 component in association with the RN condition (latency = 136 ms) showed significantly stronger field strengths over the left hemisphere. Control experiments revealed this lateralization effect to be related to noise rather than pitch processing. Furthermore, subtle noise variations interacted with signal periodicity. Obviously, thus, complex task demands such as RN encoding give rise to functional segregation of auditory processing across the two hemispheres (left hemisphere: noise, right hemisphere: periodicity representation). The observed noise/periodicity interactions, furthermore, might reflect pitch-synchronous spectral evaluation at the level of the left supratemporal plane, triggered by right-hemisphere representation of signal periodicity. PMID- 15351631 TI - Selective rotation of egocentric spatial representation following right putaminal hemorrhage. AB - Although the role of frontoparietal cortex in spatial egocentric processing is well established, recent animal-lesion and human functional imaging studies have suggested that the neostriatum may also be a critical modulator in the processing of body-centred spatial orientation. We describe here a patient with right putamen-centred hemorrhage who exhibited a consistent counterclockwise rotation of approximately 90 degrees when drawing and writing from memory. A more detailed assessment with a series of representational clock tests demonstrated that the rotation was present only in tasks requiring the use of egocentric cues. In the absence of external cues the patient would adopt and maintain a stable but incorrectly-oriented egocentric representation of the imagined or recollected object. By contrast, performance could be rectified by presentation of correctly oriented stimuli. These findings suggest that the putamen is part of a circuit underlying egocentric, as opposed to allocentric, representation of space in humans. PMID- 15351632 TI - Performance on the IOWA card task by adolescents and adults. AB - Performance on the Iowa Card Task (ICT) is impaired in several populations that demonstrate poor decision-making and risk-taking including patients with damage to the ventromedial prefrontal cortex. For some individuals, adolescence is a period of poor decision-making and risk-taking. Adolescence is also a period of neuroanatomic and neurochemical remodeling. There is evidence that neuropsychological functions of the prefrontal cortex change during adolescence; however, decision-making, as measured by the ICT, has not been systematically studied in adolescents over the age of 14 years. In the present study, 60 participants (30 female) in each of grades 6-12, and 60 young adults were administered the Iowa Card Task, the Wisconsin Card Sorting Task (WCST), a survey of impulsivity and excitement-seeking, and a quantity-frequency index of substance use. Results showed significant and steady improvement on the Iowa Card Task from the 6th grade to adulthood. Performance on the Iowa Card Task was not correlated with measures of impulsivity, excitement-seeking or reported individual substance use. There was one indication that poly-substance use was negatively correlated with performance on the ICT. Performance on the WCST was not significantly correlated with performance on the Iowa Card Task or any other measure. Across age, males and females had a different response pattern on the ICT. Females tended to choose cards associated with both immediate wins and with long-term outcome. Males tended to choose on the basis of long-term outcome. Results are discussed in terms of adolescent brain development and decision making differences between males and females. PMID- 15351633 TI - Hemispheric integration and differences in perception of a line-motion illusion in the divided brain. AB - Five people lacking the corpus callosum (two callosotomized, three with agenesis of the corpus callosum) and neurologically normal subjects were shown vertical lines that appeared instantaneously between pairs of rectangles in one or other visual field. When one of the rectangles flashed prior to the presentation of the line, and the line was in the same visual field, all subjects perceived the line as spreading from the flashed rectangle to the other. Normal subjects and one of the callosotomized subjects showed a slight but significant right visual-field advantage, perhaps reflecting a left-hemispheric superiority in processing rapid temporal events. The illusion was also induced when the line and the flash were in opposite visual fields in one of the callosotomized, one of the acallosal subjects, and about half of the normal subjects, implying interhemispheric integration even in the absence of the corpus callosum. PMID- 15351634 TI - Deficits in long-term retention of learned motor skills in patients with cortical or subcortical degeneration. AB - We investigated the acquisition and long-term retention of new skills in patients with cortical (Alzheimer's disease, AD) and subcortical (progressive supranuclear palsy, PSP; Parkinson's disease, PD) degeneration. The motor skill task performance of the PD and PSP patients improved with training, but the improvement disappeared within a few months, whereas AD patients retained learned skills for 3-18 months. The results of our experiments show that subcortical dysfunction induces a retention deficit for newly learned motor skills. Our present study suggests that a normal striatum is necessary for the formation of long-lasting motor skills, and that the striatum plays an important role as a motor skill consolidation system. PMID- 15351635 TI - A comparative analysis of RNA targeting strategies in the thymosin beta 4 gene. AB - The thymosin beta 4 (Tbeta4) gene is of biological and pharmaceutical relevance because of its anti-inflammatory and wound-healing properties. As such, it is an example of a gene that may be targeted in immunotherapy regimens. Therefore, we have used the Tbeta4 gene to compare alternative strategies for RNA targeting, namely short hairpin (sh) RNAi versus external guide sequence (EGS)-mediated RNase P cleavage. Tbeta4 has two transcripts (UTbeta4 and LTbeta4) formed by alternative splicing that differ in both expression levels and the biological activity of their encoded products. Thus, we were able to compare the capacity of shRNAi/EGS mini-genes to target molecules of high and low abundance; to specifically target alternatively spliced mRNAs; and to discriminate between very closely related alleles encoding for identical proteins. Finally, we compared transient gene knockdown in tissue culture with results in stable systems in vitro and in vivo. The data demonstrate that shRNAi and EGS can both target the Tbeta4 gene, but that the extent of RNA reduction with shRNAi ( approximately 90%) is greater. RNAi targeting shows varying efficacy against two overlapping RNAs, is largely but not completely splice form-specific, and preferentially, but not exclusively, targets a perfect-sequence match. Very high targeting achieved with an shRNAi expressed from an RNA polymerase III promoter in transient transfection was not maintained in stably transfected clones and was not efficiently transmitted through the mouse germline. These results demonstrate the versatility and the limitations of RNA targeting strategies, and suggest that particular biological and clinical needs may be best met by varying the strategy. PMID- 15351636 TI - Inhibition of the expression of the human RNase P protein subunits Rpp21, Rpp25, Rpp29 by external guide sequences (EGSs) and siRNA. AB - External guide sequences (EGSs) and siRNAs were targeted individually to the mRNA of three of the protein subunits of human RNase P, Rpp21, Rpp25 and Rpp29. The production of each of the three targets was inhibited in every specific case. In addition, some of the remaining protein subunits were also inhibited by these specific EGSs and the siRNAs. These data, in general, confirm previous results on the inhibition of a sub-group of all the protein subunits with an EGS against Rpp38. The effect of EGSs is apparent in 24 hours after transfection but the effect of siRNAs, which is comparable to the EGS data in amounts of inhibition, takes at least 48 to 96 hours to become evident. No general understanding of the mechanism of action of the siRNAs, in terms of which portion of a target mRNA they bind to for function, was apparent from the design of those used here. PMID- 15351637 TI - Transcription factors IIF and IIS and nucleoside triphosphate substrates as dynamic probes of the human RNA polymerase II mechanism. AB - The mechanism for elongation catalyzed by human RNA polymerase II (RNAP II) has been analyzed using millisecond phase transient state kinetics. Here, we apply a running start, two-bond, double-quench protocol. Quenching the reaction with EDTA indicates NTP loading into the active site followed by rapid isomerization. HCl quenching defines the time of phosphodiester bond formation. Model-independent and global kinetic analyses were applied to simulate the RNAP II mechanism for forward elongation through the synthesis of two specific phosphodiester bonds, modeling rate data collected over a wide range of nucleoside triphosphate concentrations. We report adequate two-bond kinetic simulations for the reaction in the presence of TFIIF alone and in the presence of TFIIF+TFIIS, providing detailed insight into the RNAP II mechanism and into processive RNA synthesis. RNAP II extends an RNA chain through a substrate induced-fit mechanism, termed NTP-driven translocation. After rapid isomerization, chemistry is delayed. At a stall point induced by withholding the next templated NTP, RNAP II fractionates into at least two active and one paused conformation, revealed as different forward rates of elongation. In the presence of TFIIF alone or in the presence of TFIIF+TFIIS, rapid rates are very similar; although, with TFIIF alone the complex is more highly poised for forward synthesis. Based on steady-state analysis, TFIIF was thought to suppress transcriptional pausing, but this view is misleading. TFIIF supports elongation and suppresses pausing by stabilizing the post-translocated elongation complex. When TFIIS is present, RNA cleavage and transcriptional restart pathways are supported, but TFIIS has a role in suppression of transient pausing, which is the most important contribution of TFIIS to elongation from a stall position. PMID- 15351638 TI - Complex interactions between bovine plasminogen and streptococcal plasminogen activator PauA. AB - The interactions between bovine plasminogen and the streptococcal plasminogen activator PauA that culminate in the generation of plasmin are not fully understood. Formation of an equimolar activation complex comprising PauA and plasminogen by non-proteolytic means is a prerequisite to the recruitment of substrate plasminogen; however the determinants that facilitate these interactions have yet to be defined. A mutagenesis strategy comprising nested deletions and random point substitutions indicated roles for both amino and carboxyl-terminal regions of PauA and identified further essential residues within the alpha domain of the plasminogen activator. A critical region within the alpha domain was identified using non-overlapping PauA peptides to block the interaction between PauA and bovine plasminogen, preventing formation of the activation complex. Homology modelling of the activation complex based upon the known structures of streptokinase complexed with human plasmin supported these findings by placing critical residues in close proximity to the plasmin component of the activation complex. PMID- 15351639 TI - Role of the putative structural protein Sed1p in mitochondrial genome maintenance. AB - The nuclear gene MIP1 encodes the mitochondrial DNA polymerase responsible for replicating the mitochondrial genome in Saccharomyces cerevisiae. A number of other factors involved in replicating and segregating the mitochondrial genome are yet to be identified. Here, we report that a bacterial two-hybrid screen using the mitochondrial polymerase, Mip1p, as bait identified the yeast protein Sed1p. Sed1p is a cell surface protein highly expressed in the stationary phase. We find that several modified forms of Sed1p are expressed and the largest of these forms interacts with the mitochondrial polymerase in vitro. Deletion of SED1 causes a 3.5-fold increase in the rate of mitochondrial DNA point mutations as well as a 4.3-fold increase in the rate of loss of respiration. In contrast, we see no change in the rate of nuclear point mutations indicating the specific role of Sed1p function in mitochondrial genome stability. Indirect immunofluorescence analysis of Sed1p localization shows that Sed1p is targeted to the mitochondria. Moreover, Sed1p is detected in purified mitochondrial fractions and the localization to the mitochondria of the largest modified form is insensitive to the action of proteinase K. Deletion of the sed1 gene results in a reduction in the quantity of Mip1p and also affects the levels of a mitochondrially-expressed protein, Cox3p. Our results point towards a role for Sed1p in mitochondrial genome maintenance. PMID- 15351640 TI - Characterization of hnRNP K protein-RNA interactions. AB - The heterogeneous nuclear ribonucleoprotein K protein is an RNA-binding protein found in several subcellular compartments where it is thought to be involved in signaling multiple processes that compose gene expression. K protein contains three K homology (KH) domains that mediate RNA-binding. We used a serial analysis of gene expression (SAGE)-based strategy, yeast three-hybrid screen, RNA pull down assays and computational analysis to characterize K protein-associated RNAs. We demonstrate that K protein interacts with many sense and antisense nuclear and mitochondrial transcripts through both direct and indirect binding. The highly specific direct binding of transcripts to K protein is mediated by a consensus sequence comprising three C-rich patches. Structural analysis suggests a three prong interaction model whereby each of the three KH domains binds one of the C rich patches. Genome-wide and yeast three-hybrid clone analysis revealed that these sequences are located preferentially in the 3' untranslated regions, which are known to regulate mRNA translation and processing. PMID- 15351641 TI - A conserved zinc binding domain in the largest subunit of DNA-dependent RNA polymerase modulates intrinsic transcription termination and antitermination but does not stabilize the elongation complex. AB - An evolutionarily conserved zinc-binding motif is found close to the amino terminus of the largest subunits of DNA-dependent RNA polymerases from bacteria, archaea, and eukaryotes. In bacterial RNA polymerase, this motif, the zinc binding domain, has been implicated in protein-DNA interactions that stabilize the transcription elongation complex and that occur downstream of the catalytic center. Here, we show that this view is incorrect, and instead, the zinc binding domain interacts with product RNA located upstream of the catalytic center and the RNA-DNA hybrid, a view consistent with structural studies of the elongation complex. We engineered mutations that alter or remove the zinc binding domain of Escherichia coli RNA polymerase. Several mutants, including one that lacked all four zinc ligands and another that lacked the entire domain, produced enzymes that were active in vitro and formed stable elongation complexes. However, they were defective in two functions that require interaction of polymerase with product RNA. First, they terminated less efficiently than the wild-type at intrinsic transcription terminators. Second, enzymes lacking the tip of the zinc binding domain or the zinc ligands did not antiterminate in response to an intrinsic antiterminator encoded by the put site of phage HK022. Termination, but not antitermination, was restored by the bacterial termination factor NusA. Surprisingly, a mutant that lacks the entire zinc binding domain regained a partial response to put. To account for this we suggest that put RNA interacts with an additional site in the elongation complex to mediate antitermination, and that this site is occluded by the wild-type zinc binding domain. PMID- 15351642 TI - Crystal structure of an IdeR-DNA complex reveals a conformational change in activated IdeR for base-specific interactions. AB - The iron-dependent regulator (IdeR) is an essential protein in Mycobacterium tuberculosis that regulates iron uptake in this major pathogen. Under high iron concentrations, IdeR binds to several operator regions and represses transcription of target genes. Here, we report the first crystal structure of cobalt-activated IdeR bound to the mbtA-mbtB operator at 2.75 A resolution. IdeR binds to the DNA as a "double-dimer" complex with two dimers on opposite sides of the DNA duplex with the dimer axes deviating approximately 157 degrees. The asymmetric unit contains two such double-dimer complexes with a total molecular mass of 240 kDa. Two metal-binding sites are fully occupied with the SH3-like third domain adopting a "wedge" position to interact with the two other domains, and providing two ligands for the metal site 1 in all eight subunits per asymmetric unit. A putative sodium ion is observed to mediate interactions between Asp35 and DNA. There is a conformational change in the DNA-binding domain caused by a 6-9 degrees rotation of the helix-turn-helix motif with respect to the rest of the molecule upon binding to the DNA. Ser37 and Pro39 make specific interactions with conserved thymine bases while Gln43 makes non-specific contacts with different types of bases in different subunits. A "p1s2C3T4a5" base recognition pattern is proposed to be the basis for key interactions for each IdeR subunit with the DNA in the IdeR-DNA complex structure. PMID- 15351643 TI - Regulation of hem gene expression in Rhodobacter capsulatus by redox and photosystem regulators RegA, CrtJ, FnrL, and AerR. AB - Biosynthetic pathways for heme and chlorophyll share common intermediates from 5 aminolevulinic acid through protoporphyrin IX. To obtain a better understanding of how photosynthetic organisms coordinate heme and chlorophyll biosynthesis, we have undertaken detailed analysis of the expression pattern of numerous heme biosynthesis genes in the purple photosynthetic bacterium Rhodobacter capsulatus. beta-Galactosidase reporter assays demonstrated that expression of hemA, hemB, hemC, hemE and hemZ genes is elevated under conditions that give rise to elevated bacteriochlorophyll synthesis. Heme gene expression is shown to be affected by mutations in previously identified transcriptional regulators RegA, FnrL, CrtJ, and AerR, which also control expression of genes involved in bacteriochlorophyll and carotenoid synthesis, and synthesis of the apoprotein subunits of the photosynthetic and electron transport apparatus. High-resolution primer extension analysis of hem mRNA reveals the presence of numerous putative RegA, FnrL and CrtJ binding sites in several hem promoter regions. PMID- 15351644 TI - Efficient regulation of sigmaF, the first sporulation-specific sigma factor in B.subtilis. AB - Differential gene expression is established in the prespore and mother-cell compartments of Bacillus subtilis through the successive activation of a series of cell-type-specific sigma factors. Crucial to the success of this process is the control of the first prespore-specific sigma factor, sigmaF. sigmaF is regulated by the proteins SpoIIAB, SpoIIAA and SpoIIE. SpoIIAB forms an inhibitory complex with sigmaF, which can be dissociated by interaction with SpoIIAA. During this interaction SpoIIAA is phosphorylated. SpoIIE is a membrane bound phosphatase that dephosphorylates SpoIIAA, thereby re-activating it. It is not understood how sigmaF is activated specifically in the prespore but not in the mother cell. Here, we use a recently developed fluorescence spectroscopy technique to follow in real time the formation of sigmaF.SpoIIAB complexes and their dissociation by SpoIIAA. We show that complete activation of sigmaF is induced by a tenfold increase in SpoIIE activity. This result demonstrates that relatively small changes in SpoIIE activity, which could arise from asymmetric septation, can achieve the all-or-nothing response in sigmaF activity required by the cell. For long-term sigmaF activation, we find that sustained SpoIIE activity is required to counteract the activity of SpoIIAB. Even though the continual phosphorylation and dephosphorylation of SpoIIAA by these two enzymes will expend some ATP, the formation of SpoIIAA.SpoIIAB.ADP complexes greatly diminishes the rate of the phosphorylation reaction, and thus minimizes the wastage of energy. These features provide a very efficient system for regulating sigmaF. PMID- 15351645 TI - The 1.3 A crystal structure of human mitochondrial Delta3-Delta2-enoyl-CoA isomerase shows a novel mode of binding for the fatty acyl group. AB - The crystal structure of Delta3-Delta2-enoyl-CoA isomerase from human mitochondria (hmEci), complexed with the substrate analogue octanoyl-CoA, has been refined at 1.3 A resolution. This enzyme takes part in the beta-oxidation of unsaturated fatty acids by converting both cis-3 and trans-3-enoyl-CoA esters (with variable length of the acyl group) to trans-2-enoyl-CoA. hmEci belongs to the hydratase/isomerase (crotonase) superfamily. Most of the enzymes belonging to this superfamily are hexamers, but hmEci is shown to be a trimer. The mode of binding of the ligand, octanoyl-CoA, shows that the omega-end of the acyl group binds in a hydrophobic tunnel formed by residues of the loop preceding helix H4 as well as by side-chains of the kinked helix H9. From the structure of the complex it can be seen that Glu136 is the only catalytic residue. The importance of Glu136 for catalysis is confirmed by mutagenesis studies. A cavity analysis shows the presence of two large, adjacent empty hydrophobic cavities near the active site, which are shaped by side-chains of helices H1, H2, H3 and H4. The structure comparison of hmEci with structures of other superfamily members, in particular of rat mitochondrial hydratase (crotonase) and yeast peroxisomal enoyl CoA isomerase, highlights the variable mode of binding of the fatty acid moiety in this superfamily. PMID- 15351646 TI - Conformational changes of troponin C within the thin filaments detected by neutron scattering. AB - Regulation of skeletal and cardiac muscle contraction is associated with structural changes of the thin filament-based proteins, troponin consisting of three subunits (TnC, TnI, and TnT), tropomyosin, and actin, triggered by Ca2+ binding to TnC. Knowledge of in situ structures of these proteins is indispensable for elucidating the molecular mechanism of this Ca2+-sensitive regulation. Here, the in situ structure of TnC within the thin filaments was investigated with neutron scattering, combined with selective deuteration and the contrast matching technique. Deuterated TnC (dTnC) was first prepared, this dTnC was then reconstituted into the native thin filaments, and finally neutron scattering patterns of these reconstituted thin filaments containing dTnC were measured under the condition where non-deuterated components were rendered "invisible" to neutrons. The obtained scattering curves arising only from dTnC showed distinct difference in the absence and presence of Ca2+. These curves were analyzed by model calculations using the Monte Carlo method, in which inter-dTnC interference was explicitly taken into consideration. The model calculation showed that in situ radius of gyration of TnC was 23 A (99% confidence limits between 22 A and 23 A) and 24 A (99% confidence limits between 23 A and 25 A) in the absence and presence of Ca2+, respectively, indicating that TnC within the thin filaments assumes a conformation consistent with the extended dumbbell structure, which is different from the structures found in the crystals of various Tn complexes. Elongation of TnC by binding of Ca2+ was also suggested. Furthermore, the radial position of TnC within the thin filament was estimated to be 53 A (99% confidence limits between 49 A and 57 A) and 49 A (99% confidence limits between 44 A and 53 A) in the absence and presence of Ca2+, respectively, suggesting that this radial movement of TnC by 4A is associated with large conformational changes of the entire Tn molecule by binding of Ca2+. PMID- 15351647 TI - Helical order in tarantula thick filaments requires the "closed" conformation of the myosin head. AB - Myosin heads are helically ordered on the thick filament surface in relaxed muscle. In mammalian and avian filaments this helical arrangement is dependent on temperature and it has been suggested that helical order is related to ATP hydrolysis by the heads. To test this hypothesis, we have used electron microscopy and image analysis to study the ability and temperature dependence of analogs of ATP and ADP.Pi to induce helical order in tarantula thick filaments. ATP or analogs were added to rigor myofibrils or purified thick filaments at 22 degrees C and 4 degrees C and the samples negatively stained. The ADP.Pi analogs ADP.AlF4 and ADP.Vi, and the ATP analogs ADP.BeFx, AMPPNP and ATPgammaNH2, all induced helical order in tarantula thick filaments, independent of temperature. In the absence of nucleotide, or in the presence of ADP or the ATP analog, ATPgammaS, there was no helical ordering. According to crystallographic and tryptophan fluorescence studies, all of these analogs, except ATPgammaS and ADP, induce the "closed" conformation of the myosin head (in which the gamma phosphate pocket is closed). We suggest that helical order requires the closed conformation of the myosin head but is not dependent on the hydrolysis of ATP. PMID- 15351648 TI - Crystal structure of human triggering receptor expressed on myeloid cells 1 (TREM 1) at 1.47 A. AB - The triggering receptor expressed on myeloid cells (TREM) family of single extracellular immunoglobulin receptors includes both activating and inhibitory isoforms whose ligands are unknown. TREM-1 activation amplifies the Toll-like receptor initiated responses to invading pathogens allowing the secretion of pro inflammatory chemokines and cytokines. Hence, TREM-1 amplifies the inflammation induced by both bacteria and fungi, and thus represents a potential therapeutic target. We report the crystal structure of the human TREM-1 extracellular domain at 1.47 A resolution. The overall fold places it within the V-type immunoglobulin domain family and reveals close homology with Ig domains from antibodies, T-cell receptors and other activating receptors, such as NKp44. With the additional use of analytical ultracentrifugation and 1H NMR spectroscopy of both human and mouse TREM-1, we have conclusively demonstrated the monomeric state of this extracellular ectodomain in solution and, presumably, of the TREM family in general. PMID- 15351649 TI - Diversity in structure and function of the Ets family PNT domains. AB - The PNT (or Pointed) domain, present within a subset of the Ets family of transcription factors, is structurally related to the larger group of SAM domains through a common tertiary arrangement of four alpha-helices. Previous studies have shown that, in contrast to the PNT domain from Tel, this domain from Ets-1 contains an additional N-terminal helix integral to its folded structure. To further investigate the structural plasticity of the PNT domain, we have used NMR spectroscopy to characterize this domain from two additional Ets proteins, Erg and GABPalpha. These studies both define the conserved and variable features of the PNT domain, and demonstrate that the additional N-terminal helix is also present in GABPalpha, but not Erg. In contrast to Tel and Yan, which self associate to form insoluble polymers, we also show that the isolated PNT domains from Ets-1, Ets-2, Erg, Fli-1, GABPalpha, and Pnt-P2 are monomeric in solution. Furthermore, these soluble PNT domains do not associate in any pair-wise combination. Thus these latter Ets family PNT domains likely mediate interactions with additional components of the cellular signaling or transcriptional machinery. PMID- 15351650 TI - Crystal structure of the molecular chaperone HscA substrate binding domain complexed with the IscU recognition peptide ELPPVKIHC. AB - HscA, a specialized bacterial Hsp70-class molecular chaperone, interacts with the iron-sulfur cluster assembly protein IscU by recognizing a conserved LPPVK sequence motif. We report the crystal structure of the substrate-binding domain of HscA (SBD, residues 389-616) from Escherichia coli bound to an IscU-derived peptide, ELPPVKIHC. The crystals belong to the space group I222 and contain a single molecule in the asymmetric unit. Molecular replacement with the E.coli DnaK(SBD) model was used for phasing, and the HscA(SBD)-peptide model was refined to Rfactor=17.4% (Rfree=21.0%) at 1.95 A resolution. The overall structure of HscA(SBD) is similar to that of DnaK(SBD), although the alpha-helical subdomain (residues 506-613) is shifted up to 10 A relative to the beta-sandwich subdomain (residues 389-498) when compared to DnaK(SBD). The ELPPVKIHC peptide is bound in an extended conformation in a hydrophobic cleft in the beta-subdomain, which appears to be solvent-accessible via a narrow passageway between the alpha and beta-subdomains. The bound peptide is positioned in the reverse orientation of that observed in the DnaK(SBD)-NRLLLTG peptide complex placing the N and C termini of the peptide on opposite sides of the HscA(SBD) relative to the DnaK(SBD) complex. Modeling of the peptide in the DnaK-like forward orientation suggests that differences in hydrogen bonding interactions in the binding cleft and electrostatic interactions involving surface residues near the cleft contribute to the observed directional preference. PMID- 15351651 TI - Folding kinetics of an alpha helical membrane protein in phospholipid bilayer vesicles. AB - We report a detailed kinetic study of the folding of an alpha-helical membrane protein in a lipid bilayer environment. SDS denatured bacteriorhodopsin was folded directly into phosphatidylcholine lipid vesicles by stopped-flow mixing. The folding kinetics were monitored with millisecond time resolution by time resolving changes in protein fluorescence as well as in the absorption of the retinal chromophore. The kinetics were similar to those previously reported for folding bacteriorhodopsin in detergent or lipid micelles, except for the presence of an additional apoprotein intermediate. We suggest this intermediate is a result of the greater internal two-dimensional pressure present in these lipid vesicles as compared to micelles. These results lay the groundwork for future studies aimed at understanding the mechanistic origin of the effect of lipid bilayer properties on protein folding. Furthermore, the use of biologically relevant phosphatidylcholine lipids, together with a straightforward rapid mixing process to initiate the folding reaction, means the method is generally applicable, and thus paves the way for an improved understanding of the in vitro folding of transmembrane alpha-helical proteins. PMID- 15351652 TI - Controlling the folding efficiency of an integral membrane protein. AB - Research into the folding mechanisms of integral membrane proteins lags far behind that of water-soluble proteins, to the extent that the term protein folding is synonymous with water-soluble proteins. Hydrophobic membrane proteins, and particularly those with transmembrane alpha-helical motifs, are frequently considered too difficult to work with. We show that the stored curvature elastic stress of lipid bilayers can be used to guide the design of efficient folding systems for these integral membrane proteins. The curvature elastic stress of synthetic phosphatidylcholine/phosphatidylethanolamine lipid bilayers can be used to control both the rate of folding and the yield of folded protein. The use of a physical bilayer property generalises this approach beyond the particular chemistry of the lipids involved. PMID- 15351653 TI - Conformational prerequisites for formation of amyloid fibrils from histones. AB - We demonstrate that bovine core histones are natively unfolded proteins in solutions with low ionic strength due to their high net positive charge at pH 7.5. Using a variety of biophysical techniques we characterized their conformation as a function of pH and ionic strength, as well as correlating the conformation with aggregation and amyloid fibril formation. Tertiary structure was absent under all conditions except at pH 7.5 and high ionic strength. The addition of trifluoroethanol or high ionic strength induced significant alpha helical secondary structure at pH 7.5. At low pH and high salt concentration, small-angle X-ray scattering and SEC HPLC indicate the histones are present as a hexadecamer of globular subunits. The secondary structure at low pH was independent of the ionic strength or presence of TFE, as judged by FTIR. The data indicate that histones are able to adopt five different relatively stable conformations; this conformational variability probably reflects, in part, their intrinsically disordered structure. Under most of the conditions studied the histones formed amyloid fibrils with typical morphology as seen by electron microscopy. In contrast to most aggregation/amyloidogenic systems, the kinetics of fibrillation showed an inverse dependence on histone concentration; we attribute this to partitioning to a faster pathway leading to non-fibrillar self associated aggregates at higher protein concentrations. The rate of fibril formation was maximal at low pH, and decreased to zero by pH 10. The kinetics of fibrillation were very dependent on the ionic strength, increasing with increasing salt concentration, and showing marked dependence on the nature of the ions; interestingly Gdn.HCl increased the rate of fibrillation, although much less than NaCl. Different ions also differentially affected the rate of nucleation and the rate of fibril elongation. PMID- 15351654 TI - Structure of the constitutively active double mutant CheYD13K Y106W alone and in complex with a FliM peptide. AB - CheY is a member of the response regulator protein superfamily that controls the chemotactic swimming response of motile bacteria. The CheY double mutant D13K Y106W (CheY**) is resistant to phosphorylation, yet is a highly effective mimic of phosphorylated CheY in vivo and in vitro. The conformational attributes of this protein that enable it to signal in a phosphorylation-independent manner are unknown. We have solved the crystal structure of selenomethionine-substituted CheY** in the presence of its target, a peptide (FliM16) derived from the flagellar motor switch, FliM, to 1.5A resolution with an R-factor of 19.6%. The asymmetric unit contains four CheY** molecules, two with FliM16 bound, and two without. The two CheY** molecules in the asymmetric unit that are bound to FliM16 adopt a conformation similar to BeF3- -activated wild-type CheY, and also bind FliM16 in a nearly identical manner. The CheY** molecules that do not bind FliM16 are found in a conformation similar to unphosphorylated wild-type CheY, suggesting that the active phenotype of this mutant is enabled by a facile interconversion between the active and inactive conformations. Finally, we propose a ligand-binding model for CheY and CheY**, in which Ile95 changes conformation in a Tyr/Trp106-dependent manner to accommodate FliM. PMID- 15351655 TI - Aquaporin-0 membrane junctions form upon proteolytic cleavage. AB - Aquaporin-0 (AQP0), previously known as major intrinsic protein (MIP), is the only water pore protein expressed in lens fiber cells. AQP0 is highly specific to lens fiber cells and constitutes the most abundant intrinsic membrane protein in these cells. The protein is initially expressed as a full-length protein in young fiber cells in the lens cortex, but becomes increasingly cleaved in the lens core region. Reconstitution of AQP0 isolated from the core of sheep lenses containing a proportion of truncated protein, produced double-layered two-dimensional (2D) crystals, which displayed the same dimensions as the thin 11 nm lens fiber cell junctions, which are prominent in the lens core. In contrast reconstitution of full-length AQP0 isolated from the lens cortex reproducibly yielded single layered 2D crystals. We present electron diffraction patterns and projection maps of both crystal types. We show that cleavage of the intracellular C terminus enhances the adhesive properties of the extracellular surface of AQP0, indicating a conformational change in the molecule. This change of function of AQP0 from a water pore in the cortex to an adhesion molecule in the lens core constitutes another manifestation of the gene sharing concept originally proposed on the basis of the dual function of crystallins. PMID- 15351656 TI - Navigation-aided reconstruction of medial orbital wall and floor contour in cranio-maxillofacial reconstruction. AB - The reconstruction of the anterio-posterior inclination of the medial aspect of the orbital floor, despite a wide 360 degrees exposure, including coronal and conjunctival incisions, is a challenging task in severe injuries of the orbit with massive comminution and complete displacement of the medial orbital wall and orbital floor. Out of a total of 20 patients with orbital fractures, five underwent a surgical intervention of repositioning the medial aspect of the orbital floor and especially the transition area between the orbital floor and medial orbital wall, using navigation-aided procedures. Using the mirroring tool of the Stryker-Leibinger STN-system, post-operative CTs indicated an average difference of the globe position of -4.9% between the operated side and the unaffected side, depending on the position of the medial aspect of the orbital floor. Navigation-aided procedures proved to be an essential precondition for achieving precise and predictable results in orbital reconstruction. In such cases, unlike those with an intact medial orbital wall remnant as a surgical target, bone grafts for reconstruction of the orbital floor cannot be implanted as onlay grafts. PMID- 15351657 TI - Ocular injuries caused by plastic bullet shotguns in Switzerland. AB - Five patients with blunt ocular trauma due to hard plastic shotguns used by police forces during riots presented to the Ophthalmology Department of University Hospital in Zurich, Switzerland, between December 2000 and May 2001. All five eyes suffered ocular concussion. Three of five eyes presented with severe damage to the anterior segment of the eye, two of these eyes showed combined involvement of the anterior and posterior segments. Two patients completely recovered their visual acuity in the injured eye, two reached a final visual acuity of 6/12 and in one case the injured eye was legally blinded. Three of the patients claimed to have been uninvolved bystanders at the riots. The theoretical probability of hitting the head/neck area or one of the two eyes for each shot fired at a person from different operational distances is calculated and ophthalmological and technical aspects of this special type of plastic bullet shotgun used in Switzerland are discussed. PMID- 15351658 TI - Are nasal fractures being referred appropriately from the accident and emergency department to ENT? AB - OBJECTIVE: Nasal trauma does not always require ENT clinic follow-up. We assess the appropriateness of referral to the ENT department from A&E of suspected nasal fractures. METHOD: The 342 case notes of patients referred to ENT from A&E with suspected nasal fractures were reviewed. The patients were divided into groups depending on whether or not they showed clinical features in A&E. RESULTS: Patients showing clinical features in A&E initially were significantly [P < .005] more likely to attend their clinic appointment than those without clinical features. They were also significantly more likely [P < 0.001] to undergo surgical correction of their nose. CONCLUSIONS: Suspected nasal fractures following trauma should be referred to ENT only if they show clinical features on presentation to A&E. For patients developing clinical features over the next 3 weeks a telephone number should be given to the patient for rapid access to the ENT clinic. PMID- 15351659 TI - The 'Pillow Burrito' for hand elevation. PMID- 15351660 TI - Evaluation of techniques for treating the bleeding wound. AB - BACKGROUND: Despite the fact that the urgent control of active bleeding from external body surfaces demands a basic procedure, it is puzzling that this topic has received little if any attention in modern medical research. Elastic adhesive dressing (ELAD) has been proposed for dressing the bleeding wound. METHODS AND MATERIALS: Three techniques were compared over a simulated wound in a human model. Pressures generated between the chosen dressing surface and the underlying mock wound's cutaneous surface, time consumption, convenience, satisfaction and pain were tested for each technique. RESULTS: Sixty-eight experiments were performed over nine separate anatomical sites. Average pressures for field dressing, ELAD and manual compression were 33, 88 and 180 mmHg, respectively; these differences in pressure were statistically significant. Manual pressure was equally inconvenient for both patient and caregiver. The more proximal and wider anatomical regions were more difficult and time consuming to compress. The caregivers graded ELAD the highest level of convenience and general satisfaction. CONCLUSIONS: Field bandage testing reflected its inadequacy in controlling bleeding from most body regions. The results suggest that ELAD may be the hands free technique of choice. We hope that this article will stimulate further research and elicit evidence on precisely which technique is most suitable for various anatomical location. PMID- 15351661 TI - A novel system for the irrigation of open fractures. AB - It is accepted that a good washout of contaminated wounds with copious fluids reduces the rate of infection. We describe a simple method of rapidly irrigating wounds with high volumes of fluid. PMID- 15351662 TI - Wet cement remains a poorly recognised cause of full-thickness skin burns. AB - In 1993 we published a paper stating that wet construction cement is a poorly recognised cause of full-thickness burns [Injury 24 (1993) 615]. Ten years since publication we re-evaluated cement burns to ascertain if there had been any changes in awareness and incidence. All cement related burns assessed at this unit were reviewed over a three and a half year period. Postal questionnaires were sent to all those affected. Cement burns accounted for 2.2% of referrals to the Burns Unit during this studied period as compared to 1% during the original study. Fifty-one percent of patients stated they were unaware of the risk of cement burns and took no precautions. Eighty-six percent of the burns involved the lower limb. Fifty-three percent of burns affected professional users. Fifty percent had full-thickness involvement with 21% requiring operation. Eighty-eight percent of patients did not notice any warnings on the cement bag or delivery docket and 95% felt warnings should be larger and more obvious. This study has shown that cement burns may have resulted from shortcomings in the information and warnings provided to customers by cement manufacturers. PMID- 15351663 TI - Prevention of secondary osteoporotic fractures--why are we ignoring the evidence? AB - Osteoporosis is a major problem world-wide. In the UK, it costs the NHS about pound 1.7 billion per annum. Admission to orthopaedic units for treatment and rehabilitation with fractured necks of femur makes up a large proportion of the cost. The evidence suggests simple, cost-effective treatments can reduce fractures and re-fractures attributable to osteoporosis by up to 50%, and easy to follow guidelines exist. Our study, of a typical large district general hospital suggests that ideally placed orthopaedic surgeons are however not taking the opportunity to start these treatments or offer advice about this common condition. Commencing these treatments could prevent more than 7000 fractures per year in the UK. PMID- 15351664 TI - Basal fractures of the femoral neck: intra- or extra-capsular. AB - The method of treatment and outcome for 83 patients presenting with a basal fracture of the femoral neck were determined. This represented 2.3% of all hip fractures admitted to one institution. The mean age of patients was 80 years, 25% were male. Seventy-one fractures were treated with a sliding hip screw, one of which cut-out. No other failures of fixation occurred or re-operations were required. Two fractures were fixed with cancellous screws, one healed in varus and the other fixation failed requiring re-operation. Six were managed with an arthroplasty and four treated conservatively. At 1-year from injury 29% of patients had died. For the survivors 87% were able to return home and only 6% had significant residual pain. PMID- 15351665 TI - Prediction of fixation failure after sliding hip screw fixation. AB - Cut-out of the lag screw is the commonest cause of fixation failure after sliding hip screw fixation of extracapsular hip fracture. A number of technical aspects of surgery have been used to asses the risk of cut-out. This study was to determine which of these indicators was the most reliable predictor of cut-out. The anterior-posterior and lateral post-operative radiographs of 23 cases of cut out were compared with those of 77 cases of uneventful fracture healing. The tip apex distance with correction for magnification was found to show the most significant difference between patients with cut-out against those without (P = 0.001), followed by the lag screw position on the lateral radiographs (P = 0.0095 and 0.014), reduction of the fracture on the anterior-posterior radiograph (P = 0.011 and 0.016) and the uncorrected tip-apex distance (P = 0.019). We recommend that for audit and research purposes the corrected tip-apex distance, fracture reduction and implant positioning methods should be used. For routine clinical practice, the uncorrected tip to apex distance, which is sum of the distance from the tip of the lag screw to the apex of the femoral head on anterior-posterior and lateral radiograph, and fracture reduction angle on the anterior-posterior radiograph are recommended. PMID- 15351666 TI - DHS osteosynthesis for stable pertrochanteric femur fractures with a two-hole side plate. AB - The sliding hip screw is the implant of choice for the operative treatment of stable pertrochanteric femur fractures (AO classification 31-A.1). For this indication, a four-hole side plate with four bicortical screws is widely used to allow full weight bearing immediately after operation, but scientific support for the need of such a long side plate is not available. A shorter side plate is potentially less invasive. Therefore, we retrospectively evaluated all 148 consecutive patients (median age 80 years) with a stable pertrochanteric femur fracture who were treated between 1995 and 2001 with a dynamic hip screw (DHS) and a short (two-hole) side plate and immediate full weight bearing mobilisation. Although two wound hematomas and four wound infections occurred, 145 fractures healed radiologically without implant-related complications within 6 months. Two hip screws cut out of the femoral head due to a poor position. In one patient, the side plate broke out after a fall out of bed on the fifth postoperative day. Pull off of any two-hole side plate during early full weight bearing mobilisation without further trauma was not observed. We conclude that fixation of stable pertrochanteric femur fractures with a two-hole DHS is safe. The traditional use of a four-hole DHS plate for this indication is therefore 'over-treatment' since it is more invasive. PMID- 15351667 TI - Retrospective analysis of factors influencing the operative result after percutaneous osteosynthesis of intracapsular femoral neck fractures. AB - A retrospective analysis of 104 patients with an intracapsular femoral neck fracture, treated with closed reduction and internal fixation showed that the Garden classification, the age of the patient and the quality of reduction and fixation are predictors for the final outcome. Fractures with a Garden classification of 1 and 2 have significantly better results than Garden 3 and 4 (P < 0.01). An unsuccessful reduction of the fracture and a poor surgical fixation technique had significant effect on the outcome (P = 0.01 and 0.02, respectively). If internal fixation is considered, the reduction and the technique of fixation should be perfect. PMID- 15351668 TI - Screening time for extra-capsular proximal femoral fracture fixation; the difference between extra-medullary and intra-medullary implant usage. AB - The aim of this study was to compare the fluoroscopic screening time used for treatment of fractures of the trochanteric region of the femur using two different implant systems. Data were collected from 277 proximal femoral fracture fixations. A dynamic hip screw (DHS) was used in 145, and an intra-medullary hip screw (IMHS) was used in 132. There was no difference between the two groups with respect to age, gender or side. Altogether, there were 42% two parts, 35% were three parts and 23% were four parts extra-capsular neck fractures. There was no statistical difference in ionising radiation exposure in closed reduction of these fractures regardless of the fracture configuration or surgical experience of the surgeon. The mean screening time to implant a DHS in two part fractures was 0.48 min, for three part fractures it was 0.45 min and for four part fractures it was 0.46 min. The mean screening time to implant IMHS was 1.02 min for two part fractures, 0.96 min for three part fractures and 1.03 min for four part fractures. These differences were statistically significant (P < or = 0.05). PMID- 15351669 TI - Mortality following surgery for undisplaced intracapsular hip fractures. AB - PURPOSE: The aim of the study was to evaluate the mortality following the operative treatment of undisplaced subcapital fracture of the hip by internal fixation (with three lag screws) or hemiarthroplasty. METHODS: A prospective audit of all patients admitted with hip fracture was undertaken at the university hospital in Nottingham. An independent research assistant collected data on a standardised questionnaire. Mortality was calculated from data received from National office of Statistics allowing 100% 1-year follow up for mortality statistics. RESULTS: One hundred and sixty patients were admitted with undisplaced intracapsular fracture of the hip. Twenty-one patients had non operative management and were excluded from the results. One hundred and thirty nine patients had surgical treatment. Mean age of patients was 78 years. Twenty nine patients had hemiarthroplasty and 110 patients underwent internal fixation of their fractures. There was no significant difference between the two groups for age, sex, mobility, residential status, co-morbidity and cognitive state. There was a significant difference in mortality between the two operated groups at 1 month and 1 year after the operation. Six patients (21%) died after hemiarthroplasty in the first month while there were only two (2%) deaths in the internal fixation group (P < 0.001). At 1 year from operation, 11 patients (38%) from the hemiarthroplasty group and 17 patients (16%) from the internal fixation group died (P = 0.0072). The re-operation rate within 1 year was higher for the internal fixation group (n = 8; 7.2%) than the hemiarthroplasty group (n = 1; 3%). CONCLUSIONS: There is significant increase in mortality when undisplaced intracapsular hip fractures are treated by hemiarthroplasty as compared to internal fixation and we would not recommend it for these fractures. PMID- 15351670 TI - Critical radiological analysis after Austin Moore hemiarthroplasty. AB - The aim of this study is to investigate the causes of prosthesis loosening in patients treated with Austin Moore hemiarthroplasty (AMA). The clinical and radiological outcomes were documented in a quantitative manner after 7 years follow-up of 144 patients. At the time of final follow-up, 52 patients had died and 48 patients were lost to follow-up, leaving a total of 44 patients for analysis. Immediate post-operative X-rays were studied for the initial alignment of prosthesis, the fit of the prosthesis and the degree of osteoporosis. X-rays on latest follow-up were studied for evidence of loosening. All patients were assessed clinically with the hip score of hospital for special surgery. It was found that hip pain was significantly related to subsidence and pivoting of the prosthesis (P = 0.014 and 0.035, respectively). Significant increase in subsidence was noted if the stem of prosthesis was not fitting well within the shaft of femur (P = 0.006). When the patient was younger than 73 years old at the time of operation, there was more subsidence of the prosthesis at the final follow-up (P = 0.001). It was concluded that the fill of AMA within the shaft of femur should be greater than 70% to avoid early loosening. Relatively younger patients with acute fracture of the neck of femur should be treated by methods other than cementless AMA. PMID- 15351671 TI - Hip fractures in centenarians. AB - The centenarian population is increasing yet there is little about their morbidity and mortality rates following hip fracture. The aim was to review centenarians treated for proximal femoral fractures in Edinburgh describing treatment outcomes in relation to mortality, walking ability and residential status comparing centenarians with a the more typical hip fracture population. In this retrospective review, 18 centenarians sustaining hip fractures in Edinburgh between 1998 and 2002 were compared to 18 randomly selected "normal" hip fracture patients aged 75-83 years. Centenarian in-hospital, 1 and 4 month mortality was 11.1, 33.3 and 50%, respectively, versus 0, 0 and 5.6% in the normal group. Centenarian 4 month mortality was significantly greater than that of the normal group (Fisher's Exact Test, P = 0.00723). A total of 22.2% of centenarians regained pre-fracture walking ability compared to 58.8% of the normal patients. A total 28.6% of centenarians could continue living independently post-fracture compared to 69.2% of the normal group. PMID- 15351672 TI - Cephalomedullary interlocked nail for ipsilateral hip and femoral shaft fractures. AB - A retrospective study of the management of 23 cases of ipsilateral hip and femoral shaft fractures, between January 1998 and December 2001, is presented. All except two cases were managed by a single implant, i.e. reconstruction nail. There was delayed diagnosis of femoral neck fracture in two cases where the "miss a nail" technique was used for fixation of the femoral neck fracture. All patients managed by reconstruction nail were simultaneously operated on for both fractures and operative treatment was executed as early as the general condition of the patient permitted. Delay in treatment was generally because of the associated injuries (head, chest or abdominal). There were 22 males and 1 female patient with an average age of 34.5 years. Average follow-up was 30.9 months. There was one case of non-union of the femoral neck fracture, one case of avascular necrosis and one femoral neck fracture united in varus. There were four cases of non-union and six cases of delayed union of femoral shaft fractures. Mean time for union of the femoral neck fracture was 15 weeks and for the shaft fracture was 22 weeks. In this series femoral shaft fracture determined the total union period. Complications involving the femoral shaft fracture were more common than those related to femoral neck fractures. Shaft complications were more manageable with or without secondary procedures as compared to femoral neck complications, which usually require more extensive procedures. This stresses the need to realise the significance and seriousness of both components of this complex injury, in evaluation, management and post-operative care. We conclude that, though technically demanding, reconstruction nail is an acceptable alternative for management of concomitant fractures of the femoral neck and shaft with acceptable rates of complications and good results. PMID- 15351673 TI - Origami in dynamic hip screw surgery. AB - The insertion of a dynamic hip screw requires assessment of the native angle of the femoral neck. The two commonly used angles of DHS insertion are 135 and 150 degrees. We present a simple way of judging whether a 135 or 150 degrees angle plate is more suitable in the preoperative plan by using one or two sheets of A4 or letter paper, respectively. PMID- 15351674 TI - The blast component of firework injuries--not to be underestimated. PMID- 15351675 TI - Not just another 'head lac'...low-velocity, penetrating intra-cranial injuries: a case report and review of the literature. PMID- 15351676 TI - Surgical management of traumatic luxation of the ruptured eyeball. PMID- 15351677 TI - Fatal retropharyngeal haematoma secondary to blunt trauma. PMID- 15351678 TI - Combined anterior and posterior stabilisation for treating an unstable cervical spine fracture in a patient with long standing ankylosing spondylitis. PMID- 15351679 TI - Simultaneous post-traumatic 'criss cross' dislocation of hip joints--one anterior and other posterior. PMID- 15351680 TI - Unusual presentation of sciatica in a 14-year-old girl. AB - The sciatic nerve can be compressed by a variety of causes, while intervertebral disc herniation is the most common cause of sciatica [Surg. Neurol. 46 (1996) 14], other documented causes include, infection, neoplasm, degenerative disease of a spine, congenital anomalies and traumatic posterior hip dislocation [BMJ 287 (1983) 157]. Sciatic neuropathy in children is uncommon. We present an unusual case of sciatic nerve compression in a 14-year-old-girl that was caused by an avulsion fracture of the ischial tuberosity. The compression was relieved by surgical excision of the avulsed ischial tuberosity. PMID- 15351681 TI - Simultaneous bilateral fractures of femoral neck in children--mechanism of injury. PMID- 15351682 TI - Cryopreservation of viable hepatocyte monolayers in cryoprotectant media with high serum content: metabolism of testosterone and kaempherol post cryopreservation. AB - Little work in the literature focuses on the cryopreservation of primary hepatocytes as monolayer cultures, yet this technique offers many distinct advantages over other cryopreservation systems, including high recovery, high post-thaw nutrient penetration, and low numbers of trapped dead cells. This article investigates the cryopreservation of primary rat hepatocytes at -78 degrees C attached as monolayers to collagen coated culture dishes, and describes efforts to increase post-thaw viability and function through manipulation of the freeze/thaw protocol. Different concentrations of foetal calf serum (FCS) with 10% (v/v) dimethyl sulphoxide (ME2SO) were tested as cryopreservation media, and high cryoprotectant serum levels were found to be important in maintaining membrane integrity and function in the cryopreserved rat hepatocyte monolayer cultures. Cultures cryopreserved with 90% (v/v) FCS plus 10% (v/v) ME2SO maintain 79.7+/-6.5% of the monolayer area as viable cells with normal morphology (by image analysis), 112.7+/-14.2% protein concentration, 55.4+/-4.2% carboxyfluorescein diacetate de-acetylation, 27.2+/-7.5% kaempherol glucuronidation (a measure of UDP-glucuronosyl transferase activity), and 39.3+/ 7.3% testosterone hydroxylation (a measure of cytochrome P-450 activity) compared with non-cryopreserved controls. This method of cryopreservation may provide a simple, convenient means of long-term storage of hepatocytes for in vitro metabolism studies. PMID- 15351683 TI - Studies on chilling sensitivity of zebrafish (Danio rerio) oocytes. AB - Human activity in the last few decades has had a devastating effect on the diversity of fresh water and marine fish. Further decline of fish population may have serious economic and ecological consequences. One of the most promising techniques to preserve fish population is to cryopreserve their germ cells. Cryopreservation has been successfully applied to fish sperm of many species, but there has been no success with fish embryo cryopreservation and fish oocyte cryopreservation has never been studied systematically. The aim of this study is to investigate the chilling sensitivity of fish oocytes. Experiments were conducted with zebrafish stage III (vitellogenic) and stage V (mature) oocytes, which were chilled at 10, 5, 0, -5 or -10 degrees C for 15 or 60 min using a low temperature bath. Control oocytes were kept at room temperature at 22 degrees C. Oocyte viability was assessed using three different methods: trypan blue staining (TB), thiazolyl blue tetrazolium bromide (MTT) staining and observation of germinal vesicle breakdown (GVBD). The results showed that zebrafish oocyte are very sensitive to chilling and their survival decreased with decreasing temperature and increasing exposure time periods. Normalised survivals assessed with TB staining after exposure to 0, -5 or -10 degrees C for 15 or 60 min were 90.1+/-6.0, 77.8+/-7.6, and 71.2+/-9.3%, and 60.2+/-3.8, 49.6+/-6.7, and 30.4+/ 3.0%, respectively. The study found that the sensitivity of viability assessment methods increase in the order of MTT < TB < GVBD. It was found that stage III oocytes were more susceptible to chilling than stage V oocytes, and that individual female had a significant influence (p < 0.0001) on oocyte chilling sensitivity. Zebrafish oocyte chilling sensitivity may also be one of the limiting factors for development of protocol of their cryopreservation. PMID- 15351684 TI - Loading red blood cells with trehalose: a step towards biostabilization. AB - A method for freeze-drying red blood cells (RBCs) while maintaining a high degree of viability has important implications in blood transfusion and clinical medicine. The disaccharide trehalose, found in animals capable of surviving dehydration can aid in this process. As a first step toward RBC preservation, we present a method for loading RBCs with trehalose. The method is based on the thermal properties of the RBC plasma membranes and provides efficient uptake of the sugar at 37 degrees C in a time span of 7 h. The data show that RBCs can be loaded with trehalose from the extracellular medium through a combination of osmotic imbalance and the phospholipid phase transition, resulting in intracellular trehalose concentrations of about 40 mM. During the loading period, the levels of ATP and 2,3-DPG are maintained close to the levels of fresh RBCs. Increasing the membrane fluidity through the use of a benzyl alcohol results in a higher concentration of intracellular trehalose, suggesting the importance of the membrane physical state for the uptake of the sugar. Osmotic fragility data show that trehalose exerts osmotic protection on RBCs. Flow cytometry data demonstrate that incubation of RBCs in a hypertonic trehalose solution results in a fraction of cells with different complexity and that it can be removed by washing and resuspending the RBCs in an iso-osmotic medium. The data provide an important first step in long-term preservation of RBCs. PMID- 15351685 TI - Sperm membrane fatty acid composition in the Eastern grey kangaroo (Macropus giganteus), koala (Phascolarctos cinereus), and common wombat (Vombatus ursinus) and its relationship to cold shock injury and cryopreservation success. AB - Marsupial spermatozoa tolerate cold shock well, but differ in cryopreservation tolerance. In an attempt to explain these phenomena, the fatty acid composition of the sperm membrane from caput and cauda epididymides of the Eastern grey kangaroo, koala, and common wombat was measured and membrane sterol levels were measured in cauda epididymidal spermatozoa. While species-related differences in the levels of linolenic acid (18:3, n-6) and arachidonic acid (20:4, n-6) were observed in caput epididymal spermatozoa, these differences failed to significantly alter the ratio of unsaturated/saturated membrane fatty acids. However in cauda epididymidal spermatozoa, the ratio of unsaturated/saturated membrane fatty acids in koala and kangaroo spermatozoa was approximately 7.6 and 5.2, respectively; substantially higher than any other mammalian species so far described. Koala spermatozoal membranes had a higher ratio of unsaturated/saturated membrane fatty acids than that of wombat spermatozoa (t = 3.81; df = 4; p < or = 0.02); however, there was no significant difference between wombat and kangaroo spermatozoa. The highest proportions of DHA (22:6, n 3), the predominant membrane fatty acid in cauda epididymidal spermatozoa, were found in wombat and koala spermatozoa. While species-related differences in membrane sterol levels (cholesterol and desmosterol) were observed in cauda epididymidal spermatozoa, marsupial membrane sterol levels are very low. Marsupial spermatozoal membrane analyses do not support the hypothesis that a high ratio of saturated/unsaturated membrane fatty acids and low membrane sterol levels predisposes spermatozoa to cold shock damage. Instead, cryogenic tolerance appears related to DHA levels. PMID- 15351686 TI - Cryopreservation of the endangered mahseer (Tor khudree) spermatozoa: I. Effect of extender composition, cryoprotectants, dilution ratio, and storage period on post-thaw viability. AB - Several in situ and ex situ conservation strategies have been suggested for the revival of stocks of Tor khudree (Sykes), a threatened species. Cryopreservation of spermatozoa is crucial for the conservation of stocks of endangered species so that sustainable production can be ensured. Among the different extenders, modified fish Ringer (E1) was found to be the best for cryopreservation of T. khudree spermatozoa. Extender E2 appeared the next best. Extenders based on chicken egg yolk and milk powder were found to be unsuitable for the cryopreservation of T. khudree spermatozoa. Among the cryoprotectants, dimethyl sulfoxide provided maximum protection to spermatozoa during freezing and thawing. Propylene glycol and methanol were found to be less effective. Of the four spermatozoa dilutions, 1:10, 1:15, and 1:20 showed better motility rates than 1:5. At the former dilution ratios, the motility rates which were more than 95% prior to freezing were reduced to 80-81 and 43-67%, 10 and 70 days after cryopreservation, respectively. The motility duration did not differ much with increasing storage period at all the dilution ratios. Motility rates generally decreased with an increase in frozen storage. When spermatozoa were thawed and stored at 25 degrees C for varying periods, motility percentage, and duration decreased gradually as the storage period increased; spermatozoa stored up to 40 min after thawing retained 55% motility and were motile up to 77s; these values declined further leading to the complete cessation of motility 70 min after storage. The importance of extender-cryoprotectant mixture, milt dilution, and storage period in developing a protocol for T. khudree spermatozoa cryopreservation is discussed. PMID- 15351687 TI - Effect of cryopreservation on expression of Th1 and Th2 cytokines in blood mononuclear cells from patients with different cytokine profiles, analysed with three common assays: an overall decrease of interleukin-4. AB - Studies on cytokine expression in blood cells are commonly performed on cryopreserved cells. Previous studies show that cryopreservation affects cytokine expression, but the findings are not consistent. This may be due to divergent effects of freezing on different cytokines, different stimuli, and different patient groups or to the use of different assays in the studies. This study was designed to investigate the effect of freezing on spontaneous, auto-antigen, allergen, and mitogen induced cytokine secretion from peripheral blood mononuclear cells from several groups of patients expressing different cytokine profiles; multiple sclerosis, atopic children, non-atopic children, and pregnant women. The expression of IFN-gamma, IL-4, IL-5, IL-9, IL-10, and IL-13 was analysed with ELISA, ELISPOT and/or real time RT-PCR. Our data provide evidence that the process of cryopreservation and thawing does affect the expression of cytokines, both at the protein and the mRNA level. Moreover, the effect varied among different cytokines, different stimuli, and different patient groups, which partly may be explained by differences in optimal freezing conditions for non activated and activated cells. An increase of allergen and PHA stimulated IFN gamma secretion in atopic children was found following cryopreservation, but no such increase in auto-antigen induced IFN-gamma was seen in MS-patients. The most consistent finding was that expression of IL-4 was generally decreased in spontaneous and auto-antigen/allergen induced expression in cryopreserved cells. In conclusion, this study points out the importance of investigation of the effects of freezing for each cytokine, stimuli and patient group before using frozen cells in studies of in vitro cytokine secretion. PMID- 15351688 TI - Cryoprotectant permeability parameters for cells used in a bioengineered human corneal equivalent and applications for cryopreservation. AB - A human corneal equivalent is being developed with applications in pharmaceutical testing and biomedical research, but the distribution of this engineered tissue, depends on successful cryopreservation. Cryopreservation of tissues depends on the presence of cryoprotectants, their addition and removal, and exposure to conditions during freezing and thawing, all of which depend on cellular membrane permeabilities to water and cryoprotectant. This study defines the permeability properties that define the rate of water and cryoprotectant movement across the plasma membrane of isolated human corneal endothelial, keratocyte, and epithelial cells. Cells were transferred from isotonic conditions (300 mosm/kg) to 0.5, 1, or 2 M dimethyl sulfoxide and propylene glycol solutions at constant temperature, and cell volumes monitored using an electronic particle counter. Histograms describing cell volume changes over time after cryoprotectant exposure allowed calculation of hydraulic conductivity (Lp), cryoprotectant permeability (Ps), and the reflection coefficient (sigma). Experimental values for Lp and Ps at 4, 13, 22, and 37 degrees C were used to determine the Arrhenius activation energy (Ea). Defining the permeability parameters and temperature dependencies allows simulation of responses of human corneal cells to addition and removal of cryoprotectants and to freezing conditions, allowing amount of supercooling, intracellular electrolyte concentration, and intracellular cryoprotectant concentration to be calculated. Simulations also show that the constituent cells in the bioengineered cornea respond differently to addition and removal of cryoprotectants and to freezing. This study has defined the requirements during cryopreservation for the corneal cells; future work will define the matrix requirements which will allow the development of a cryopreservation protocol. PMID- 15351689 TI - High ice nucleation temperature of zebrafish embryos: slow-freezing is not an option. AB - Although fish embryos have been used in a number of slow-freezing cryopreservation experiments, they have never been successfully cryopreserved. In part this is because little is known about whether ice forms within the embryo during the slow-freezing dehydration process. Therefore, we examined the temperature of intraembryonic ice formation (T(IIF)) and the temperature of extraembryonic ice formation (T(EIF)), using a cryomicroscope. We used both unmodified zebrafish embryos and those with water channels (aquaporin-3 or AQP3) inserted into their membranes to increase permeability to water and cryoprotectants, examined at 100% epiboly to the 6-somite stage. In these experiments we examined: (1) the spontaneous freezing of (external) solutions; (2) the spontaneous freezing of solutions containing embryos; (3) the effect of preloading the embryos with cryoprotectants on T(IIF); (4) whether preloading the embryos with cryoprotectant helps in survival after nucleating events in the solution; and (5) the damaging effects of extracellular nucleation events versus solution toxicity on the embryos. The solutes alone (embryo medium--EM, sucrose culture medium, 1 M propylene glycol in EM, and 1 M propylene glycol in a sucrose culture medium) froze at -14.9 +/- 1.1, -17.0 +/- 0.3, -17.8 +/- 1.0, and -17.7 +/- 1.4, respectively. There was no difference amongst these means (P > 0.05), thus adding cryoprotectant did not significantly lower the nucleation point. Adding embryos (preloaded with cryoprotectant or not) did not change the basic freezing characteristics of these solutes. In all these experiments, (T(EIF)) equaled (T(IIF)), and there was no difference in the freezing point of the solutions with or without the embryos (P > 0.05). Additionally, there was no difference in the freezing characteristics of embryos with and without aquaporins (P > 0.05). The formation of intraembryonic ice was lethal to the zebrafish embryos in all cases. But this lethal outcome was not related to solution injury effects, because 88-98% of embryos survived when exposed to a higher solute concentration with no ice present. Taken together, these data suggest that slow freezing is not a suitable option for zebrafish embryos. The mechanism of this high temperature nucleation event in zebrafish embryos is still unknown. PMID- 15351690 TI - Activation of AMP-dependent protein kinase by hypoxia and hypothermia in the liver of frog Rana perezi. AB - We have investigated different signaling molecules that could be activated by temperature acclimation and hypoxia, using an experimental approach consisting in submerging frogs in a water-filled box maintained at 2-4 degrees C at ambient oxygen levels or supplied with 98% N2:2% CO2 for normoxia or hypoxia conditions, respectively. The results obtained showed no significant changes in the expression of heat shock protein 70. The phosphorylation state of AMP-dependent activated protein kinase, the down-stream component of a protein kinase cascade that acts as an intracellular energy sensor, was significantly increased in both experimental conditions, showing higher values in the absence of oxygen. Similarly, the phosphorylation state of one of its known substrates, elongation factor 2, was also increased, consistent with the arrest of protein synthesis. These results point out an important role of this kinase, adjusting the rates of ATP-consuming and ATP-generating pathways, in the survival strategies to hypoxia and hypothermia. PMID- 15351691 TI - Inter-sample variability in post-thaw human spermatozoa. AB - Semen cryopreservation is a useful tool for preserving fertility in men who have been diagnosed with cancer and will undergo chemotherapy, radiotherapy or testicular surgery. Semen is also commonly cryopreserved prior to its use in assisted reproductive techniques such as in vitro fertilization and intracytoplasmic sperm injection. The post-thaw quality of banked sperm can vary, which may negatively affect fertilization rates. The objective of our study was to assess the pre-freeze and post-thaw variability of sperm parameters in patients who used our sperm banking services. Multiple samples obtained after a short period of sexual abstinence were examined for variation in sperm characteristics. Semen samples showed a high degree of post-thaw inter-sample variability in sperm motility, motion characteristics, and percentage cryosurvival rate compared with the pre-freeze inter-sample variability. Further research is necessary to understand the mechanism(s) responsible for this variability. This may also assist clinicians utilize semen samples with optimum semen quality in ART procedures. PMID- 15351692 TI - Cell-type-specific activation of mitogen-activated protein kinases in PAN-induced progressive renal disease in rats. AB - We examined the time-course activation and the cell-type specific role of MAP kinases in puromycin aminonucleoside (PAN)-induced renal disease. The maximal activation of c-Jun-NH2-terminal kinase (JNK), extracellular signal regulated kinase (ERK), and p38 MAP kinase was detected on Days 52, 38, and 38 after PAN treatment, respectively. p-JNK was localized in mesangial and proximal tubular cells at the early renal injury. It was expressed, therefore, in the inflammatory cells of tubulointerstitial lesions. While, p-ERK was markedly increased in the glomerular regions and macrophages p-p38 was observed in glomerular endothelial cells, tubular cells, and some inflammatory cells. The results show that the activation of MAP kinases in the early renal injury by PAN-treatment involves cellular changes such as cell proliferation or apoptosis in renal native cells. The activation of MAP kinases in infiltrated inflammatory cells and fibrotic cells plays an important role in destructive events such as glomerulosclerosis and tubulointerstitial fibrosis. PMID- 15351693 TI - TRIM45, a novel human RBCC/TRIM protein, inhibits transcriptional activities of ElK-1 and AP-1. AB - The tripartite motif (TRIM) proteins play important roles in a variety of cellular functions including cell proliferation, differentiation, development, oncogenesis, and apoptosis. In this study, we report the identification and characterization of the human tripartite motif-containing protein 45 (TRIM45), a novel member of the TRIM family, from a human embryonic heart cDNA library. TRIM45 has a predicted 580 amino acid open reading frame, encoding a putative 64 kDa protein. The N-terminal region harbors a RING finger, two B-boxes, and a predicted alpha-helical coiled-coil domain, which together form the RBCC/TRIM motif found in a large family of proteins, whereas the C-terminal region contains a filamin-type immunoglobulin (IG-FLMN) domain. Northern blot analysis indicates that TRIM45 is expressed in a variety of human adult and embryonic tissues. In the cell, TRIM45 protein is expressed both in cytoplasm and in cell nucleus. Overexpression of TRIM45 in COS-7 cells inhibits the transcriptional activities of ElK-1 and AP-1. These results suggest that TRIM45 may act as a new transcriptional repressor in mitogen-activated protein kinase signaling pathway. PMID- 15351694 TI - Reverse endocytosis of transmembrane ephrin-B ligands via a clathrin-mediated pathway. AB - Eph/ephrin receptors and ligands mediate cell-cell interaction through reciprocal signaling upon juxtacrine contact, and play a critical role in embryonic patterning, neuronal targeting, and vascular assembly. To study transmembrane ephrin-B ligand trafficking, we determined the cellular localization of ephrin-B1 GFP upon engagement by EphB1. Under normal culture conditions ephrin-B1-GFP is localized to the plasma membrane, mostly at the lateral cell borders. Addition of soluble EphB1-Fc receptor induces ephrin-B1-GFP clustering on the cell surface and subsequent internalization, as judged by biochemical studies, electron microscopy, and co-localization with endosomal markers. A dominant-negative mutant of dynamin or potassium depletion blocks ephrin-B1 endocytosis. These results suggest that ephrin-B1 internalization is an active receptor-mediated process that utilizes the clathrin-mediated endocytic pathway. PMID- 15351695 TI - Crystal structure of an acidic platelet aggregation inhibitor and hypotensive phospholipase A2 in the monomeric and dimeric states: insights into its oligomeric state. AB - Phospholipases A2 belong to the superfamily of proteins which hydrolyzes the sn-2 acyl groups of membrane phospholipids to release arachidonic acid and lysophospholipids. An acidic phospholipase A2 isolated from Bothrops jararacussu snake venom presents a high catalytic, platelet aggregation inhibition and hypotensive activities. This protein was crystallized in two oligomeric states: monomeric and dimeric. The crystal structures were solved at 1.79 and 1.90 angstroms resolution, respectively, for the two states. It was identified a Na+ ion at the center of Ca2+-binding site of the monomeric form. A novel dimeric conformation with the active sites exposed to the solvent was observed. Conformational states of the molecule may be due to the physicochemical conditions used in the crystallization experiments. We suggest dimeric state is one found in vivo. PMID- 15351696 TI - Anti-sense expression of a metallopeptidase gene enhances nuclear entry of HBV DNA. AB - Although several putative hepatitis B virus (HBV) receptors have been identified, none of them is capable of initiating HBV replication in a non-permissive human cell line. Using an Epstein-Barr virus-based extrachromosomal replication system, we have screened through a human liver cDNA library and successfully identified a clone capable of facilitating nuclear transport of HBV-DNA during the early phase of HBV infection. This clone contained a cDNA encoding a metallopeptidase-like protein in anti-sense orientation. Pretreatment of naive HepG2 cells with 1,10 phenanthroline, an inhibitor for liver metallopeptidases, led to nuclear entry of HBV-DNA after HBV infection. However, cccDNA was still undetectable in the nuclei, indicating other cellular factors required to complete the replication cycle were still missing. Our present data suggest that in the initial stage of HBV infection, liver metallopeptidase constitutes a barrier for effective nuclear entry of HBV genomic DNA. Attenuation of metallopeptidase activity may facilitate HBV infection. PMID- 15351697 TI - Expression analysis of members of the neuronal calcium sensor protein family: combining bioinformatics and Western blot analysis. AB - We have used in silico mining of public databases (NCBI UniGene and NCI SAGE Anatomic Viewer) as a tool to obtain the tissue distribution pattern of three members of the neuronal calcium sensor protein family, namely VILIP-1, hippocalcin, and NCS-1 in humans. The theoretical human mRNA expression profile of the calcium sensor proteins derived from expressed sequence tag (EST) and serial analysis of gene expression (SAGE) data was compared with expression data from human tissues obtained by Western blot analysis. Since the EST databank searches do not yet give comparable results for rat which is often used as model animal, we have also analyzed the protein expression in rat tissues. Similar to the human expression profile in rat tissues calcium sensor proteins are mainly detected in the nervous system, but the data consistently implicated the additional expression in peripheral tissues with remarkable differences between the calcium sensor proteins. PMID- 15351698 TI - Intracellular mechanisms mediating the anti-apoptotic action of gastrin. AB - We previously reported that gastrin (G17) inhibits apoptosis of AR4-2J pancreatic adenocarcinoma cells, through the activation of Akt. We dissected the mechanisms responsible for this effect. D2, a CCKB receptor antagonist, inhibited G17 induction of Akt phosphorylation, measured by Western blots with anti-phospho-Akt antibodies. The intracellular calcium chelator BAPTA-AM, but not the PKC inhibitor GF109203X, blocked G17 induction of Akt. G17 stimulated BAD phosphorylation, measured by both Western blots with anti-phospho-BAD antibodies and by in vitro Akt kinase assays using recombinant BAD as substrate. G17 also induced FOXO3 phosphorylation assessed by Western blots with anti-phospho-FOXO3 antibodies, and BAPTA-AM inhibited this effect. Gastrin inhibited luciferase activity in cells transfected with FOXO1 together with a vector containing insulin-responsive sequences upstream of the luciferase reporter gene. In conclusion, G17 induces Akt through activation of CCKB receptors and of intracellular calcium-dependent, PKC-independent, pathways. This effect leads to BAD phosphorylation and to forkhead transcription factors inactivation. PMID- 15351699 TI - Independence of hyperleptinemia-induced fat disappearance from thyroid hormone. AB - Sustained hyperleptinemia induced in normal rats causes the rapid disappearance of body fat. This is attributed to a marked increase in uncoupled fatty acid oxidation in the white adipocytes, which also occurs in hyperthyroidism. Because hyperleptinemic rats have normal plasma T3 or T4 levels, we tested the possibility of "localized hyperthyroidism" due to increased conversion of T4 to T3 in the adipose tissue. We therefore induced sustained hyperleptinemia in normal rats by intravenous injection of recombinant adenovirus containing the leptin cDNA (AdCMV-leptin) and measured the mRNA and the activity of enzymes involved in T4 metabolism in the disappearing fat. The epididymal fat pad remnants exhibited a decrease in mRNA of deiodinase 1 and a doubling of deiodinase 2 mRNA (p<0.05), but their enzyme activities did not differ from normoleptinemic controls. To determine if thyroid hormone was required for the fat-wasting action of hyperleptinemia, we infused AdCMV-leptin into rats made athyroid by total thyroidectomy or by methimazole therapy. The fat loss in hyperleptinemic athyroid rats was as great as in euthyroid controls. We conclude that the fat-wasting effect of sustained hyperleptinemia does not involve "local hyperthyroidism" in white adipose tissue and does not require thyroid hormone. PMID- 15351700 TI - Binding analyses of Cry1Ab and Cry1Ac with membrane vesicles from Bacillus thuringiensis-resistant and -susceptible Ostrinia nubilalis. AB - The binding properties of Bacillus thuringiensis toxins to brush border membrane vesicles of Dipel-resistant and -susceptible Ostrinia nubilalis larvae were compared using ligand-toxin immunoblot analysis, surface plasmon resonance (SPR), and radiolabeled toxin binding assays. In ligand-toxin immunoblot analysis, the number of Cry1Ab or Cry1Ac toxin binding proteins and the relative toxin binding intensity were similar in vesicles from resistant and susceptible larvae. Surface plasmon resonance with immobilized activated Cry1Ab toxin indicated that there were no significant differences in binding with fluid-phase vesicles from resistant and susceptible larvae. Homologous competition assays with radiolabeled Cry1Ab and Cry1Ac toxin and vesicles from resistant and susceptible larvae resulted in similar toxin dissociation constants and binding site concentrations. Heterologous competition binding assays indicated that Cry1Ab and Cry1Ac completely competed for binding, thus they share binding sites in the epithelium of the larval midguts of O. nubilalis. Overall, the binding analyses indicate that resistance to Cry1Ab and Cry1Ac in this Bt-resistant strain of O. nubilalis is not associated with a loss of toxin binding. PMID- 15351701 TI - Exchangeable zinc ions transiently accumulate in a vesicular compartment in the yeast Saccharomyces cerevisiae. AB - The baker's yeast Saccharomyces cerevisiae was used as a model to visualize intracellular labile zinc under conditions of nutritional zinc imbalance. Zinc specific staining was performed in yeast cells using both Zinquin fluorescence and zinc-selenium autometallography. Both techniques resulted in specific labeling of an intracellular vesicular compartment that was present in wild type cells as well as in the vacuolar Zn transporter mutants Deltazrc1 and Deltacot1. This compartment, that closely resembles mammalian zincosomes, appeared rapidly under conditions of zinc availability and was independent of endocytosis. However, persistence of the zinc loaded vesicles in nutritional zinc deficiency was dependent on the presence of functional Zrc1 and Cot1 vacuolar transporters. Overall our findings indicate that labile zinc in yeast cells enters a dynamic vesicular compartment which could represent an extremely important defence to buffer both zinc excess and deficiency. PMID- 15351702 TI - Chronic administration of NMU into the paraventricular nucleus stimulates the HPA axis but does not influence food intake or body weight. AB - Hypothalamic neuromedin U (NMU) appears to have a role in the regulation of appetite and the hypothalamo-pituitary-adrenal (HPA) axis. Acute administration of NMU into the paraventricular nuclei (iPVN) increases plasma adrenocorticotrophic hormone and corticosterone, and inhibits food intake in fasted rats. No studies have as yet investigated the chronic effects of centrally administered NMU. We investigated the effect of twice-daily iPVN injections of 0.3 nmol NMU for 7 days on food intake, body weight, the HPA axis, and behavior in freely fed rats. Chronic iPVN NMU was not associated with a decrease in food intake or body weight. Chronic iPVN NMU produced a typical behavioral response on day 1 and day 4 of the study, and resulted in the elevation of plasma corticosterone present 18 h after the final injection. These results suggest NMU may have a role in the regulation of the HPA axis and behavior. PMID- 15351703 TI - Potent hydroxyl radical-scavenging activity of drought-induced type-2 metallothionein in wild watermelon. AB - Wild watermelon (Citrullus lanatus sp.) has the ability to tolerate severe drought/high light stress conditions despite carrying out normal C3-type photosynthesis. Here, mRNA differential display was employed to isolate drought responsive genes in the leaves of wild watermelon. One of the isolated genes, CLMT2, shared significant homology with type-2 metallothionein (MT) sequences from other plants. The second-order rate constant for the reaction between a recombinant CLMT2 protein and hydroxyl radicals was estimated to be 1.2 x 10(11) M(-1) s(-1), demonstrating that CLMT2 had an extraordinary high activity for detoxifying hydroxyl radicals. Moreover, hydroxyl radical-catalyzed degradation of watermelon genomic DNA was effectively suppressed by CLMT2 in vitro. This is the first demonstration of a plant MT with antioxidant properties. The results suggest that CLMT2 induction contributes to the survival of wild watermelon under severe drought/high light stress conditions. PMID- 15351704 TI - Mg and Ca isotope fractionation during CaCO3 biomineralisation. AB - The natural variation of Mg and Ca stable isotopes of carbonates has been determined in carbonate skeletons of perforate foraminifera and reef coral together with Mg/Ca ratios to assess the influence of biomineralisation processes. The results for coral aragonite suggest its formation, in terms of stable isotope behaviour, approximates to inorganic precipitation from a seawater reservoir. In contrast, results for foraminifera calcite suggest a marked biological control on Mg isotope ratios presumably related to its low Mg content compared with seawater. The bearing of these observations on the use of Mg and Ca isotopes as proxies in paleoceanography is considered. PMID- 15351705 TI - Demethylating agent, 5-azacytidine, reverses differentiation of embryonic stem cells. AB - The de novo methylation activity is essential for embryonic development as well as embryonic stem (ES) cell differentiation, where the intensive and extensive DNA methylation was detected. In this study, we investigated the effects of a demethylating agent, 5-azacytidine (5-AzaC), on differentiated ES cells in order to study the possibility of reversing the differentiation process. We first induced differentiation of ES cells by forming embryoid bodies, and then the cells were treated with 5-AzaC. The cells showed some undifferentiated features such as stem cell-like morphology with unclear cell-to-cell boundary and proliferative responsiveness to LIF. Moreover, 5-AzaC increased the expressions of ES specific markers, SSEA-1, and alkaline phosphatase activity as well as ES specific genes, Oct4, Nanog, and Sox2. We also found that 5-AzaC demethylated the promoter region of H19 gene, a typical methylated gene during embryonic differentiation. These results indicate that 5-AzaC reverses differentiation state of ES cells through its DNA demethylating activity to differentiation related genes. PMID- 15351706 TI - Interaction of the BMPR-IA tumor suppressor with a developmentally relevant splicing factor. AB - Inactivation of bone morphogenetic protein signaling via mutation of the BMPR-IA TGF-beta superfamily type I receptor causes familial juvenile polyposis, an inherited gastrointestinal cancer predisposition syndrome. In an effort to provide new insight into the mechanism(s) of BMP-mediated tumor suppression, we employed a yeast two-hybrid screen to identify novel proteins that interact with the intracellular domain of BMPR-IA. 30/31 interacting clones encoded SAP49, a splicing factor that has been shown to be required for normal development in Caenorhabditis elegans. The remaining interacting clone was FKBP12.6, a known TGF beta type I receptor interactor. The interaction between BMPR-IA and SAP49 was confirmed via coimmunoprecipitation in human cells. Mutational analysis demonstrated that the GS domain of the receptor and the conserved proline-rich domain of SAP49 were required for the interaction. Co-localization studies suggested that the interaction may occur at the inner leaflet of the nuclear membrane. These data suggest that BMPR-IA may interact with and modulate the activity of a developmentally relevant splicing factor. PMID- 15351707 TI - Sprouty regulates cell migration by inhibiting the activation of Rac1 GTPase. AB - Sprouty (SPRY) protein negatively modulates fibroblast growth factor and epidermal growth factor actions. We showed that human SPRY2 inhibits cell growth and migration in response to serum and several growth factors. Using rat intestinal epithelial (IEC-6) cells, we investigated the involvement of the Rho family of GTPases, RhoA, Rac1, and cdc42 in SPRY2-mediated inhibition of cell migration and proliferation. The ability of TAT-tagged SPRY2 to inhibit proliferation and migration of IEC-6 cells transfected with constitutively active mutants of RhoA(G14V), Rac1(G12V), and cdc42 (F28L) was determined. Constitutively active RhoA(G14V), Rac1(G12V), or cdc42(F28L) did not protect cells from the anti-proliferative actions of TAT-SPRY2. The ability of TAT-hSPRY2 to inhibit migration was not altered by of RhoA(G14V) and cdc42(F28L). However, Rac1(G12V) obliterated the ability of SPRY2 to inhibit cell autonomous or serum induced migration. Also, the activation of endogenous Rac1 was attenuated by TAT SPRY2. Thus, SPRY2 mediates its anti-migratory actions by inhibiting Rac1 activation. PMID- 15351708 TI - ATP-binding cassette protein E is involved in gene transcription and translation in Caenorhabditis elegans. AB - ATP-binding cassette protein E (ABCE) gene has been annotated as an RNase L inhibitor in eukaryotes. All eukaryotic species show the ubiquitous presence and high degree of conservation of ABCEs, however, RNase L is present only in mammals. This indicates that ABCEs may function not only as RNase L inhibitors, but also may have other functions that have yet to be determined. As an initial investigation into the novel functions of ABCE, we characterized the gene (Y39E4B.1) in Caenorhabditis elegans by a combination of data mining and functional assays. ABCE promoters drove GFP expressions in hypoderm, pharynx, vulvae, head, and tail neurons at all developmental stages. Three genes, rpl-4, nhr-91, and C07B5.3, were previously found to interact with ABCE. Our expression data showed overlapping expression patterns of ABCE and rpl-4 and nhr-91, but not C07B5.3. RNAi against ABCE resulted in embryonic lethality and slow growth. These data suggest that ABCE protein might be involved in the control of translation and transcription, work as shuttle protein between cytoplasm and nucleus, and possibly as a nucleocytoplasmic transporter. In addition, RNAi data suggest that ABCE and NHR-91 may function in vulvae development and molting pathways in C. elegans. Furthermore, our data suggest that ABCE, along with its interacting components, functions in a well-conserved pathway. PMID- 15351709 TI - Redox modulation of tau and microtubule-associated protein-2 by the glutathione/glutaredoxin reductase system. AB - Alterations in the redox status of proteins have been implicated in the pathology of several neurodegenerative diseases including Alzheimer's and Parkinson's. We report that peroxynitrite and H2O2-induced disulfides in the porcine brain microtubule-associated proteins tau and microtubule-associated protein-2 are substrates for the glutaredoxin reductase system composed of glutathione reductase, human or Escherichia coli glutaredoxin, reduced glutathione, and NADPH. Oxidation and reduction of cysteines in tau and microtubule-associated protein-2 were quantitated by monitoring the incorporation of 5-iodoacetamido fluorescein, a thiol-specific labeling reagent. Reduction of disulfide bonds in the microtubule-associated proteins by the glutaredoxin reductase system restored their ability to promote the assembly of microtubules composed of purified porcine tubulin. Thiol-disulfide exchange between oxidized glutathione and the microtubule-associated proteins was detected by monitoring protein oxidation and was quantitated by measuring reduced glutathione by HPLC. PMID- 15351710 TI - Involvement of syntaxin 4 in the transport of membrane-type 1 matrix metalloproteinase to the plasma membrane in human gastric epithelial cells. AB - Membrane-type 1 matrix metalloproteinase (MT1-MMP) localized on the plasma membrane plays a central role in various normal biological responses including tissue remodeling, wound heeling, and angiogenesis and in cancer cell invasion and metastasis, by functioning as a collagenase and activating other matrix metalloproteinases. In order to elucidate the molecular mechanism of the MT1-MMP targeted localization on the plasma membrane, we examined the participation of syntaxin proteins in MT1-MMP intracellular transport to the plasma membrane in human gastric epithelial AGS cells. Western blotting showed that syntaxin 3 and 4 proteins, which are known to function in intracellular transport towards the plasma membrane, were expressed in AGS cells. Immunocytochemistry revealed that transient transfection of AGS cells with dominant-negative mutant syntaxin 4 decreased plasma membrane MT1-MMP expression. In contrast, transient transfection with either dominant-negative mutant syntaxin 3 or 7 did not affect MT1-MMP localization on the plasma membrane. Cell surface biotinylation assay and Matrigel chamber assay demonstrated that stable transfection with dominant negative mutant syntaxin 4 decreased the amount of MT1-MMP on the plasma membranes and inhibited the cell invasiveness. We suggest that syntaxin 4 is involved in the intracellular transport of MT1-MMP toward the plasma membrane. PMID- 15351711 TI - Inhibition of lipopolysaccharide-induced expression of inducible nitric oxide synthase by butein in RAW 264.7 cells. AB - Butein has been reported to exert anti-inflammatory effect but the possible mechanism involved is still unclear. Here, we report the inhibitory effect of butein on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) gene expression. Butein also inhibited the induction of tumor necrosis factor alpha and cyclooxygenase 2 by LPS. To further investigate the mechanism responsible for the inhibition of iNOS gene expression by butein, we examined the effect of butein on LPS-induced nuclear factor-kappaB (NF-kappaB) activation. The LPS-induced DNA binding activity of NF-kappaB was significantly inhibited by butein, and this effect was mediated through inhibition of the degradation of inhibitory factor-kappaB and phosphorylation of Erk1/2 MAP kinase. Furthermore, increased binding of the osteopontin alphavbeta3 integrin receptor by butein may explain its inhibitory effect on LPS-mediated NO production. Taken together, these results suggest that butein inhibits iNOS gene expression, providing possible mechanisms for its anti-inflammatory action. PMID- 15351712 TI - Immune receptors for polysaccharides from Ganoderma lucidum. AB - This study was designed to identify and characterize the immune receptors for polysaccharides from Ganoderma lucidum, a Chinese medicinal fungus that exhibits anti-tumor activities via enhancing host immunity. We herein demonstrate that G. lucidum polysaccharides (GLPS) activated BALB/c mouse B cells and macrophages, but not T cells, in vitro. However, GLPS was unable to activate splenic B cells from C3H/HeJ mice that have a mutated TLR4 molecule (incapable of signal transduction) in proliferation assays. Rat anti-mouse TLR4 monoclonal antibody (Ab) inhibited the proliferation of BALB/c mouse B cells under GLPS stimulation. Combination of Abs against mouse TLR4 and immunoglobulin (Ig) achieved almost complete inhibition of GLPS-induced B cell proliferation, implying that both membrane Ig and TLR4 are required for GLPS-mediated B cell activation. In addition, GLPS significantly inhibited the binding of mouse peritoneal macrophages with polysaccharides from Astragalus membranaceus, which is known to bind directly with TLR4 on macrophage surface. Moreover, GLPS induced IL-1beta production by peritoneal macrophages from BALB/c, but not C3H/HeJ, mice, suggesting that TLR4 is also involved in GLPS-mediated macrophage activation. We Further identified a unique 31 kDa serum protein and two intracellular proteins (ribosomal protein S7 and a transcriptional coactivator) capable of binding with GLPS in co-precipitation experiments. Our results may have important implications for our understanding on the molecular mechanisms of immunopotentiating polysaccharides from traditional Chinese medicine. PMID- 15351713 TI - Nitric oxide regulates interactions of PMN with human brain microvessel endothelial cells. AB - The hypothesis that the NO/cGMP pathway modulates PMN adhesion to human brain microvessel endothelial cells (HBMEC) was examined. Human PMN were incubated with resting or TNF-alpha-treated endothelial monolayers, and adhesion was quantified by light microscopy. TNF-alpha upregulated PMN adhesion in a time-dependent manner. Treatment of HBMEC with the NO donors SNP and DETA NONOate for 4 or 24 h decreased PMN adhesion. This was completely reversed by the guanylyl cyclase inhibitor ODQ, while addition of a cGMP agonist (8-Br-cGMP) decreased PMN adhesion. NO donors did not affect the levels of E-selectin or ICAM-1 in HBMEC. However, pre-treatment of PMN with NO donors or 8-Br-cGMP decreased their adhesion to recombinant E-selectin and ICAM-1, suggesting an effect of NO on PMN. These findings indicate that NO modulates PMN-HBMEC interactions through cGMP and decreases the binding of PMN to the adhesion molecules E-selectin and ICAM-1. PMID- 15351714 TI - The N-terminus of yeast peptide: N-glycanase interacts with the DNA repair protein Rad23. AB - Yeast peptide: N-glycanase (PNGase) is involved in the proteasomal degradation of misfolded glycoproteins where it interacts with the DNA repair protein Rad23 as first detected in a yeast two-hybrid assay and subsequently confirmed by biochemical in vivo analyses. Limited proteolysis of PNGase with trypsin led to the removal of both an N-terminal and a C-terminal stretch. Based on these truncations the N-terminal region of yeast PNGase was identified as being responsible for binding to Rad23. Secondary structure predictions of this region suggest that it is composed of a single, solvent-exposed alpha-helix. The interaction between PNGase and Rad23 was studied using surface plasmon resonance revealing an equilibrium binding constant of approximately 2.5 microM. The oligomeric nature of Rad23 was also investigated using sedimentation equilibrium analysis. Although Rad23 exists as a dimer in solution, the monomeric form of Rad23 associates with a PNGase monomer in a 1:1 stoichiometric ratio. PMID- 15351715 TI - Allogenic antibody-mediated identification of head and neck cancer antigens. AB - Recently, we described a new target-identification technology, autoantibody mediated identification of antigens (AMIDA). AMIDA takes advantage of autologous serum autoantibodies to identify disease-associated antigens. Here, we evaluated the allogenic variant of AMIDA (allo-AMIDA), using permanent cancer cell lines as an antigen-pool rather than primary biopsy samples. Twelve different proteins were retrieved exclusively with antibodies from cancer patients, but not from healthy donors. The expression of three of these antigens, e-FABP, hnRNP H, and Grb2, was evaluated in more detail. All three proteins were strongly overexpressed in primary carcinomas and metastases thereof, as compared to healthy epithelium. Additionally, serum reactivity against e-FABP was detected in 20% of cancer patients but only 2% of healthy volunteers. In summary, we demonstrate that permanent cancer cell lines represent a reliable source for tumour-associated antigens. Moreover, we show that allo-AMIDA is suitable for the identification of tumour-specific antigens overcoming the limitations of autologous screening techniques. PMID- 15351716 TI - Catalytic superoxide scavenging by metal complexes of the calcium chelator EGTA and contrast agent EHPG. AB - Metal ion chelators widely used in experimental protocols and clinical diagnosis are generally assumed to be inert. We previously reported that the ubiquitous chelator EDTA has high levels of superoxide suppressing activity. Here, we report that the common chelators calcium chelator EGTA and contrast agent EHPG have significant activities in suppressing superoxide levels depending on the nature of metal ion chelated. The most active species is Mn(II)-EGTA which exhibited an IC50 value of 0.19 microM for superoxide destruction. In addition, IC50 values for Mn(II)-EHPG and 2Cu(II)-EGTA were 0.69 and 0.60 microM, respectively. In conclusion, Mn(II) and Cu(II) complexes of the common chelators EGTA and EHPG exhibit considerable superoxide scavenging activities. Caution should be employed in their use in biological systems where superoxide has a key role and they may be useful for the development of catalytic anti-oxidants. PMID- 15351717 TI - Differential cooperation between dHAND and three different E-proteins. AB - dHAND is a transcription factor belonging to the class B basic helix-loop-helix protein family and is expressed during embryogenesis in the heart, branchial arches, limb buds, and neural crest derivatives. Despite much study, the molecular mechanisms involved in the regulation of dHAND activity are not well understood. We therefore carried out yeast two-hybrid screening using full-length dHAND as bait, which led to identification of several dHAND-binding proteins, including three E-proteins: E2A, ME2, and ALF1. Subsequent analysis revealed that although their heterodimerization and transcriptional activities were similar, dHAND/E-protein heterodimers bind to an E-box element with differing affinities, suggesting they have distinct DNA binding specificities. Moreover, in situ hybridization showed that E-protein genes are expressed fairly ubiquitously among embryonic tissues, including the branchial arches and limb buds. By contrast, little signal was detected in the heart, suggesting that dHAND complexes with partners other than E-proteins in cardiac tissue. PMID- 15351718 TI - Accumulation of pathogenic DeltamtDNA induced deafness but not diabetic phenotypes in mito-mice. AB - Mito-mice carrying various proportions of deletion mutant mtDNA (DeltamtDNA) were generated by introduction of the DeltamtDNA from cultured cells into fertilized eggs of C57BL/6J (B6) strain mice. Great advantages of mito-mice are that they share exactly the same nuclear-genome background, and that their genetic variations are restricted to proportions of pathogenic DeltamtDNA. Since accumulation of DeltamtDNA to more than 75% induced respiration defects, the disease phenotypes observed exclusively in mito-mice carrying more than 75% DeltamtDNA should be due to accumulated DeltamtDNA. In this study, we focused on the expressions of hearing loss and diabetic phenotypes, since these common age associated abnormalities have sometimes been reported to be inherited maternally and to be associated with pathogenic mutant mtDNAs. The results showed that accumulation of exogenously introduced DeltamtDNA was responsible for hearing loss, but not for expression of diabetic phenotypes in mito-mice. PMID- 15351719 TI - Structural evidence for guanidine-protein side chain interactions: crystal structure of CutA from Pyrococcus horikoshii in 3 M guanidine hydrochloride. AB - This study was carried out to investigate the structural perturbation of the protein's local structure by the denaturants under non-denaturing conditions. Crystal structure of CutA from an archaeon Pyrococcus horikosii (PhoCutA), a heavy-metal binding protein, was determined at 1.6-angstroms resolution in the presence of 3 M guanidine HCl (GdnHCl). Native PhoCutA has a large number of short intramolecular hydrogen bonds and salt bridges on the protein surface, of which greater than 90% of hydrogen bonds and all salt bridges were retained in 3 M GdnHCl. Hydrogen bonds that disappeared in the GdnHCl crystal structure were mainly located on the protein surface, especially around the structurally perturbed loop, suggesting interactions between peptide groups and GdnHCl. Only a few GdnH+ ions were observed in the crystal structure, although none at the surface, of the protein. Two GdnH+ ions were observed in the center of the trimeric structure, replacing water molecules, and were hydrogen bonded with Asp84 and Asp86 of each chain. The exterior loop from Tyr39 to Lys44, including Trp40-Trp41, was perturbed structurally. Decreases in temperature factors were observed in beta strand 5 and the N terminus of helix 3. These results suggest the specific bindings of GdnH+ with some acidic residues and the non-specific bindings around Trp residues and peptide groups on the protein surface and that binding of GdnHCl to the native protein is limited, resulting in local structural perturbation. PMID- 15351720 TI - Proinflammatory effects of pancreatic elastase are mediated through TLR4 and NF kappaB. AB - Pancreatic elastase has been implicated in the pathophysiology of severe acute pancreatitis, characterized by systemic inflammatory response, distant organ failure, and high mortality. Here we show that pancreatic elastase activates transcription factors NF-kappaB, AP-1, and NFAT in human myeloid cells (U-937 and THP-1) in culture. Pancreatic elastase also induces TNF-alpha secretion and increased expression of CD11b in THP-1 cells which can be inhibited by neutralizing anti-Toll-like receptor 4 (TLR4) antibodies. NF-kappaB blocking agents (MG-132, PGA1) prevented elastase-induced TNF-alpha secretion from THP-1 cells. Our results suggest that pancreatic elastase-induced proinflammatory effects are mediated by TLR4 and NF-kappaB in human myeloid cells. PMID- 15351721 TI - Role of the RuvAB protein in avoiding spontaneous formation of deletion mutations in the Escherichia coli K-12 endogenous tonB gene. AB - The endogenous tonB gene of Escherichia coli was used as a target for spontaneous deletion mutations which were isolated from ruvAB-, recG-, and ruvC- cells. The rates of tonB mutation were essentially the same in ruv+, ruvAB-, recG-, and ruvC cells. We analyzed tonB mutants by sequencing. In the ruv+, recG-, and ruvC- strains, the spectra were different from those obtained from the ruvAB- cells, where deletions dominated followed by IS insertions, base substitutions, and frameshifts, in that order. We then analyzed the tonB-trp large deletion, due to simultaneous mutations of the trp operon, and found that the frequency in ruvAB- was higher than those in ruv+, recG-, and ruvC- cells. To characterize deletion formation further, we analyzed all the tonB mutants from one colicin plate. Seven deletions were identified at five sites from the 45 tonB mutants of ruv+ cells and 24 deletions at 11 sites from the 43 tonB mutants of ruvAB- cells. Thus, the ruvAB- strain is a deletion mutator. We discuss the role of RuvAB in avoiding deletions. PMID- 15351722 TI - Polyamines regulate eukaryotic initiation factor 4E-binding protein 1 gene transcription. AB - Difluoromethylornithine-induced polyamine depletion produced a significant fall in the rate of 4E-BP1 gene transcription in IEC-6 cells, without a change in stability of the 4E-BP1 message. The effect was reversed by the addition of exogenous putrescine. Decreased 4E-BP1 gene transcription produced a concomitant fall in steady-state concentration of the 4E-BP1 protein. Segments of the 4E-BP1 gene 5' flanking sequence were inserted into a GFP reporter construct. While all the segments containing the first 500 nucleotides 5' to exon 1 were capable of driving GFP expression, two regions (between -2465 and -1965, and between -896 and 511) did so in a polyamine-dependent manner. Steady-state concentration of ornithine decarboxylase (ODC), the first enzyme in the polyamine biosynthetic pathway, was increased in response to polyamine depletion. These data provide a mechanism by which polyamines affect transcription of the 4E-BP1 gene, which in turn affect translation of ODC and perhaps other cap-dependent proteins. PMID- 15351723 TI - HLA class I signal transduction is dependent on Rho GTPase and ROK. AB - Chronic rejection is the major limitation to long-term allograft survival. HLA class I signaling pathways have been implicated in this process because ligation of class I molecules by anti-HLA antibodies (Ab) initiates intracellular signals in smooth muscle cells (SMC) and endothelial cells (EC) that synergize with growth factor receptors to elicit cell survival and proliferation. Anti-HLA Ab mediate cell proliferation and survival through a focal adhesion kinase dependent pathway that requires the integrity of the actin cytoskeleton. In this study, we investigated the role of Rho and Rho-kinase (ROK) in class I signal transduction. We show that class I ligation results in activation of Rho and increased stress fiber formation. In addition, inhibitors of Rho GTPase and ROK block HLA class I mediated tyrosyl phosphorylation of paxillin and FAK, central elements of the focal adhesion signaling complex. These results suggest that HLA class I-induced signaling in EC is dependent on Rho GTPase and ROK. PMID- 15351724 TI - Cellular prion protein regulates intracellular hydrogen peroxide level and prevents copper-induced apoptosis. AB - The function of cellular prion protein (PrPC), which is a copper binding protein, remains unclear. To elucidate the mechanisms in which PrPC is involved in neuroprotection, we compared death signals in prion protein gene-deficient (Prnp /-) primary cerebellar granular neurons (CGNs) to those with wild-type (WT) CGNs. When copper was exposed to these CGNs, ZrchI, and Rikn Prnp-/- CGNs were more sensitized and underwent apoptotic cell death more readily than WT CGNs. Furthermore, the level of intracellular hydrogen peroxide (H2O2) in WT CGNs increased by copper toxicity, whereas those in ZrchI and Rikn Prnp-/- CGNs did not. These results suggest that PrPC modulates the intracellular H2O2 level as a copper-binding protein to protect CGNs from apoptotic cell death possibly due to inhibiting a Fenton reaction. PMID- 15351725 TI - Identification of an osteogenic protein-1 responsive element in the aggrecan promoter. AB - Previous studies have demonstrated that osteogenic protein-1 (OP-1), a member of the bone morphogenetic protein (BMP) subfamily of the TGF-beta superfamily, stimulates aggrecan synthesis. To investigate transcriptional regulation of the aggrecan gene by OP-1, we constructed a clone containing a 1 kb region of the 5' upstream sequence of the mouse aggrecan gene fused to the promoter-less luciferase reporter gene in pGL2-Basic vector. A series of promoter deletion constructs were also generated. Studies of the promoter activity of these DNA constructs in transient-transfected cells revealed that deletion of a 33 bp region rendered the promoter unresponsive to OP-1, BMP-6, and CDMP-1 without affecting BMP-2 and TGF-beta1 responsiveness. Thus, the expression of the mouse aggrecan gene in response to BMPs appears to be the result of a unique combination of different cis-acting elements. PMID- 15351726 TI - Cloning and characterization of the promoter of human Wnt inhibitory factor-1. AB - Wnt inhibitory factor-1 (WIF-1) is a secreted antagonist of Wnt signaling and functions by directly binding to Wnt ligands in the extracellular space. Here we report the identification of the 5' promoter region (approximately 1.5 kb) of the human WIF-1 gene. Functional analysis of this region shows that a whole fragment displays high basal promoter activity in different cell lines, while the truncated forms do not, indicating that integrity of the WIF-1 promoter region may be important for WIF-1 activity. Moreover, we found that the expression level of beta-catenin in cancer cell lines correlates with the WIF-1 promoter activity, suggesting that the WIF-1 promoter may be regulated by the Wnt/beta-catenin pathway and may function in a negative feedback manner. Our results also suggest that a methylated CpG island, which we observed recently in human lung cancer, lies within the functional WIF-1 promoter region and therefore bears the importance of the methylation-status of this CpG island as an important key in Wnt activation in human cancer. PMID- 15351727 TI - Stress regulation of sulfotransferases in male rat liver. AB - Sulfotransferase (SULT) catalyzed sulfation is responsible for hormone regulation and xenobiotic detoxification. Induction of SULTs by various hormones has been reported. Stress regulation of SULTs has not been reported, however. Here we report that rat liver SULTs can be regulated by physical stress (forced running, EX) and chemical stress (the organophosphorus pesticide parathion, PS). Both EX and PS increased rat liver phenol-sulfating SULT1A1 and hydroxysteroid-sulfating SULT2A1 activities. The increase in SULT1A1 activity did not correlate with protein (Western blot) or mRNA (RT-PCR) results but correlated well with increased non-protein soluble thiols. This suggests a possible Cys modification mechanism for stress regulation of SULT1A1. In vitro studies on GSH/GSSG effects on SULT1A1 activity support this conclusion. In contrast, SULT2A1 activity following physical or chemical stress treatments correlated well with protein and mRNA levels. This suggests a stress regulation mechanism of SULT2A1 at the gene transcription level, possibly occurring via hormones. PMID- 15351728 TI - A novel IKKbeta inhibitor stimulates adiponectin levels and ameliorates obesity linked insulin resistance. AB - Adiponectin is an anti-diabetic and anti-atherogenic hormone that is exclusively secreted from fat cells. Serum adiponectin levels are reduced in obese patients and obese model mice, despite increased adipose tissue mass. Elucidation of the mechanism(s) by which plasma adiponectin levels are decreased in obese and diabetic patients would provide insight into the cause of obesity-induced diabetes and the development of therapeutic advances. In the present study, the regulation of adiponectin secretion was investigated using 3T3-L1 adipocytes and a diabetic-/obese-mouse model. A novel insulin sensitizer, IkappaB kinase beta (IKKbeta) inhibitor, ameliorated insulin resistance and up-regulated plasma levels of adiponectin without producing a significant change in body weight in KKAy mice that were fed a high-fat diet. The IKKbeta inhibitor cancelled the TNFalpha-mediated down-regulation of adiponectin secretion and simultaneously up regulated the phosphorylation of Akt in 3T3-L1 adipocytes. Using dominant negative mutants of Akt or PKClambda (downstream effectors of phosphoinositide 3 kinase), insulin-stimulated Akt activity was found to be important in the regulation of adiponectin secretion by insulin in 3T3-L1 adipocytes. These observations suggest that "insulin-stimulated Akt activity in adipocytes" may play an important role in the regulation of adiponectin secretion. PMID- 15351729 TI - Monocarboxylate transporters in subsarcolemmal and intermyofibrillar mitochondria. AB - Whether subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondria contain monocarboxylate transporters (MCTs) is controversial. We have examined the presence of MCT1, 2, and 4 in highly purified SS and IMF mitochondria. These mitochondria were not contaminated with plasma membrane, sarcoplasmic reticulum or endosomal compartments, as the marker proteins for these sub-cellular compartments (Na+-K+-ATPase, Ca2+-ATPase, and the transferrin receptor) were not present in SS or IMF mitochondria. MCT1, MCT2, and MCT4 were all present at the plasma membrane. However, MCT1 and MCT4 were associated with SS mitochondria. In contrast, the IMF mitochondria were completely devoid of MCT1 and MCT4. However, MCT2 was associated with both SS and IMF mitochondria. These observations suggest that SS and IMF mitochondria have different capacities for metabolizing monocarboxylates. Thus, the controversy as to whether mitochondria can take up and oxidize lactate will need to take account of the different distribution of MCTs between SS and IMF mitochondria. PMID- 15351730 TI - Genome comparison of a novel foot-and-mouth disease virus with other FMDV strains. AB - The genome of a novel foot-and-mouth disease virus, HKN/2002, was 8104 nucleotides (nt) in length (excluding the poly(C) tract and poly(A) tail) and was composed of a 1042-nt 5'-untranslated region (UTR), a 6966-nt open reading frame, and a 93-nt 3'-UTR. Genome sequences of HKN/2002 and other known FMDV strains were compared. The VP1, VP2, and VP3-based neighbor-joining (NJ) trees were divided into distinct clusters according to different serotypes, while other region-based NJ trees exhibited some degree of intercross among serotypes. Mutations in HKN/2002 were revealed, including frequent deletions and insertions in the G-H loop of VP1, and deletion involving 10 amino acid residues in the 3A protein. An evolutionary relationship of HKN/2002 with an Asian FMDV lineage isolated from a Hong Kong swine host in 1970 was postulated. A 43-nt deletion identified in the 5'-UTR of HKN/2002 possibly contributed to the loss of one pseudo-knot domain. PMID- 15351731 TI - In vitro inhibition of severe acute respiratory syndrome coronavirus by chloroquine. AB - We report on chloroquine, a 4-amino-quinoline, as an effective inhibitor of the replication of the severe acute respiratory syndrome coronavirus (SARS-CoV) in vitro. Chloroquine is a clinically approved drug effective against malaria. We tested chloroquine phosphate for its antiviral potential against SARS-CoV-induced cytopathicity in Vero E6 cell culture. Results indicate that the IC50 of chloroquine for antiviral activity (8.8 +/- 1.2 microM) was significantly lower than its cytostatic activity; CC50 (261.3 +/- 14.5 microM), yielding a selectivity index of 30. The IC50 of chloroquine for inhibition of SARS-CoV in vitro approximates the plasma concentrations of chloroquine reached during treatment of acute malaria. Addition of chloroquine to infected cultures could be delayed for up to 5h postinfection, without an important drop in antiviral activity. Chloroquine, an old antimalarial drug, may be considered for immediate use in the prevention and treatment of SARS-CoV infections. PMID- 15351732 TI - XBP-1 increases ERalpha transcriptional activity through regulation of large scale chromatin unfolding. AB - Human X box binding protein 1 (XBP-1) is a transcription factor essential for hepatocyte growth, the differentiation of plasma cells, and the unfolded protein response. Recently, we have demonstrated that two forms of XBP-1, XBP-1S, and XBP 1U, enhance estrogen receptor alpha (ERalpha)-dependent transcriptional activity in a ligand-independent manner. However, how XBP-1S and XBP-1U regulate ERalpha transcriptional activity remains unknown. Here, we report that XBP-1S and XBP-1U induce large-scale chromatin unfolding by targeting the XBP-1 proteins to an amplified, lac operator-containing chromosome region in mammalian cells. This unfolding activity maps to the transactivation domains of XBP-1S and XBP-1U. Wild type XBP-1S and XBP-1U, but not the mutants that completely abolished the ERalpha transcriptional activation, increased the chromatin unfolding activity of ERalpha. These data identify a novel function of XBP-1 and suggest that regulation of large-scale chromatin unfolding by XBP-1 may be responsible for the enhancement of ERalpha transcriptional activity. PMID- 15351733 TI - Double-stranded siRNA targeted to the huntingtin gene does not induce DNA methylation. AB - RNA interference is an evolutionarily conserved mechanism of post-transcriptional gene silencing. Small interfering RNAs (siRNA) of 21-23 nucleotides generated from processing double-stranded RNA (dsRNA) by ribonuclease III, Dicer, are widely used for selective sequence-specific gene silencing in a broad range of organisms. In plants, siRNA is associated with de novo RNA-directed DNA methylation (RdDM) at the homologous target genomic region. To examine RdDM in somatic cells, human glioblastoma cell lines were treated with siRNAs homologous to the human huntingtin gene responsible for Huntington's disease. Methylation of CpG dinucleotides in the plasmid vectors expressing the dsRNAs and homologous genomic region was investigated by bisulfite-mediated genomic sequencing. Target regions of the siRNA in the huntingtin gene showed no significant change in the pattern of DNA methylation, and no CpG methylation was observed on the plasmid vectors. These results indicate that siRNA is not directly linked to DNA methylation at the target huntingtin genomic locus in human cells. PMID- 15351734 TI - Gentamicin binds to the lectin site of calreticulin and inhibits its chaperone activity. AB - Recently, it became clear that aminoglycoside antibiotics affect protein-protein interactions involving protein disulfide isomerase as well as protein synthesis in the endoplasmic reticulum. In this study, we used affinity column chromatography to screen gentamicin-binding proteins in microsomes derived from bovine kidney in order to learn about the possible mechanisms of gentamicin associated nephrotoxicity. One of the gentamicin-binding proteins was identified as calreticulin (CRT) by N-terminal amino acid sequence analysis. Interestingly, gentamicin inhibited the chaperone and oxidative refolding activities of CRT when N-glycosylated substrates such as alpha1-antitrypsin and alpha-mannosidase were used as substrates, but it did not inhibit the chaperone activity of CRT when unglycosylated citrate synthase was used. Moreover, CRT suppressed the aggregation of deglycosylated and denatured alpha-mannosidase, but gentamicin did not inhibit its chaperone activity. Experiments with domain mutants suggest that the lectin site of CRT is the main target for gentamicin binding and that binding of gentamicin to this site inhibits the chaperone activity of CRT. PMID- 15351735 TI - The cloning of cyclin B3 and its gene expression during hormonally induced spermatogenesis in the teleost, Anguilla japonica. AB - We cloned cyclin B1, B2, and B3 cDNAs from the eel testis. Northern blot analysis indicated that these cyclin B mRNAs were expressed and increased from day 3 onward after the hormonal induction of spermatogenesis, and that cyclin B3 was most dominantly expressed during spermatogenesis. In situ hybridization showed that cyclin B1 and B2 were present from the spermatogonium stage to the spermatocyte stage. On the other hand, cyclin B3 mRNA was present only in spermatogonia. Although mouse cyclin B3 is expressed specifically in the early meiotic prophase, these results indicate that eel cyclin B3 expression is limited during spermatogenesis to spermatogonia, but is not present in spermatocytes. These facts together suggest that eel cyclin B3 is specifically involved in spermatogonial proliferation (mitosis), but not in meiosis. PMID- 15351736 TI - Ku80 is required but not sufficient for Galpha13-mediated endodermal differentiation in P19 embryonic carcinoma cells. AB - We have shown that a constitutively active Galpha13 (Galpha13Q226L) induces differentiation in P19 embryonic carcinoma cells to an endodermal phenotype. In this report, we demonstrate that Ku, a heterodimer of p80 (Ku80) and p70 (Ku70), is upregulated in P19 cells overexpressing Galpha13Q226L. Ku is the regulatory subunit of the DNA-dependent protein kinase and is primarily involved in DNA repair and recombination. Ku80 also is a somatostatin receptor. We show that while overexpression of Ku80 drastically reduced P19 cell proliferation, it was not sufficient to induce endodermal differentiation. However, coexpression of Galpha13Q226L and an antisense Ku80 abrogated the retarded growth rate and endodermal differentiation observed in cells expressing only Galpha13Q226L. Overexpression of Galpha13Q226L or Ku80 downregulated RNA polymerase I-mediated transcriptional activity and overexpression of antisense Ku80 restored the activity to control level. These results suggest that Ku80 is required for Galpha13-mediated endodermal differentiation in P19 cells. PMID- 15351737 TI - Establishment of hepatitis C virus replicon cell lines possessing interferon resistant phenotype. AB - To clarify the mechanism underlying resistance to interferon (IFN) by the hepatitis C virus (HCV) in patients with chronic hepatitis, we attempted to develop an IFN-resistant HCV replicon from the IFN-sensitive 50-1 replicon established previously. By treating 50-1 replicon cells with a prolonged low-dose treatment of IFN-alpha and then transfecting the total RNA derived from the IFN alpha-treated replicon cells, we successfully obtained four clones (named 1, 3, 4, and 5) of HCV replicon cells that survived against IFN-alpha (200 IU/ml). These cloned cells were further treated with IFN-alpha or IFN-beta (increased gradually to 2000 or 1000 IU/ml, respectively). This led to four replicon cell lines (alphaR series) possessing the IFN-alpha-resistant phenotype and four replicon cell lines (betaR series) possessing the IFN-beta-resistant phenotype. Furthermore, we obtained an additional replicon cell line (alphaRmix) possessing the IFN-alpha-resistant phenotype by two rounds of prolonged treatment with IFN alpha and RNA transfection as mentioned above. Characterization of these obtained HCV replicon cell lines revealed that the betaR series were highly resistant to both IFN-alpha and IFN-beta, although the alphaR series containing alphaRmix were only partially resistant to both IFN-alpha and IFN-beta. Genetic analysis of these HCV replicons found one common amino acid substitution in the NS4B and several additional amino acid substitutions in the NS5A of the betaR series, suggesting that these genetic alterations are involved in the IFN resistance of these HCV replicons. These newly established HCV replicon cell lines possessing IFN-resistant phenotypes are the first useful tools for understanding the mechanisms by which HCV acquires IFN resistance in vivo. PMID- 15351738 TI - Bax increases the pore size of rat brain mitochondrial voltage-dependent anion channel in the presence of tBid. AB - Voltage-dependent anion channel (VDAC), Bax, and tBid play a central role in apoptosis regulation but their functioning is still very controversial. VDAC forms voltage gated pore in planar lipid bilayers, and acts as the pathway for the movement of substances in and out of the mitochondria by passive diffusion. Here we report that there is increase in the pore size of VDAC in the presence of Bax and tBid through bilayer electrophysiological experiments. We hereby hypothesize that this increase in pore size might cause swelling in the mitochondria, leading to the rupture of mitochondrial outer membrane and release of cytochrome c causing brain cell death. PMID- 15351739 TI - Pancreas developing markers expressed on human mononucleated umbilical cord blood cells. AB - Haematopoietic system represents the main source of haematopoietic stem cells and probably of multipotential adult progenitor cells and mesenchimal stem cells at first described as colony forming unit-fibroblast. Whereas there are many studies on the gene expression profile of the different precursors along their haematopoietic differentiation, few data (sometimes conflicting) have been reported about the phenotype of the cells (present in bone marrow and possibly in cord blood) able to differentiate into non-haematopoietic cells. As both postnatal bone marrow and umbilical cord blood contain nestin positive cells able to proliferate and differentiate into the main neural phenotype (neuron, astroglia and oligodendroglia) many authors considered nestin a neuroepithelial precursor marker that seems to be essential also in multipotential progenitor cells of pancreas present both in rat and in human pancreatic islets (called nestin positive islet derived progenitors). Although the importance of nestin in these cells appears to be evident, it remains yet to clarify the number and the sequential expression of the genes coding all the transcription factors essential for beta cells differentiation and therefore the conditions able to induce the expression of many important transcription factors genes such as isl-1, pax-4, pdx-1 and ngn-3. Among them pdx-1 is a gene essential for pancreas development which is able to control ngn-3 in activating the expression of other differentiation factors for endocrine cells. Here, we describe for the first time in human umbilical cord blood cells (UCB) the pattern of expression of a panel of markers (nestin, CK-8, CK-18) and transcription factors (Isl-1, Pdx-1, Pax-4, Ngn 3) considered important for beta cells differentiation. Our data demonstrate that UCB contains a cell population having a phenotype very similar to endocrine cell precursors in transition to beta cells. PMID- 15351740 TI - Functional expression of TREK-2 in insulin-secreting MIN6 cells. AB - Insulin secretion from pancreatic beta cells is partly regulated by cell membrane potential. Background K+ channels that stabilize the resting membrane potential would suppress excitability and insulin secretion. Recent studies show that members of the two-pore domain K+ (K2P) channel family behave as background K+ channels in many excitable cells. Therefore, the expression of K2P channels was studied in insulin-secreting MIN6 cells. Reverse transcriptase PCR showed that, among nine K2P channels tested, TASK-1, TASK-2, TASK-3, TREK-2, and TRESK-2 were expressed in MIN6 cells. Cell-attached recordings on MIN6 cells revealed five types of K+ channels that were open at rest. Two were ATP-sensitive and Ca2+ activated K+ channels, as judged by their sensitivity to ATP and Ca2+, respectively, and single-channel conductance. Among five K2P channels, only TREK 2 could be clearly identified in MIN6 cells. The molecular identity of two other K+ channels is not yet known. TREK-2 in MIN6 cells was activated by arachidonic acid, membrane stretch, and low pH solution (pH 5.8). Arachidonic acid increased Ba2+-sensitive whole-cell current in MIN6 cell. These results suggest that TREK-2 contributes to the background K+ conductance in MIN6 cells, and may regulate depolarization-induced secretion of insulin. PMID- 15351741 TI - SRA coactivation of estrogen receptor-alpha is phosphorylation-independent, and enhances 4-hydroxytamoxifen agonist activity. AB - The ability of steroid receptor RNA activator (SRA), an AF-1 coactivator, to contribute to differences in estrogen receptor (ER)-alpha and ERbeta transcriptional activity was tested. In transient transfections, SRA expression increased ERalpha- and ERbeta-dependent gene expression. However, when the receptors' amino-terminal A/B regions were examined as GAL4 DNA binding domain fusions, SRA enhanced the activity of GAL-ABalpha but not GAL-ABbeta. Exogenous SRA also enhanced AF-2 activity for both receptors, indicating that SRA effects are not limited to AF-1. Simultaneously mutating three phosphorylation sites within GAL-ABalpha domain only modestly reduced SRA coactivation of GAL-ABalpha, suggesting that phosphorylation does not play a major role in SRA function relative to this domain. SRA enhanced ERalpha activity stimulated by 4 hydroxytamoxifen, but was unable to convert this mixed antiestrogen to an ERbeta agonist. Thus, SRA is an ERalpha AF-1-specific coactivator that enhances the agonist activity of tamoxifen-bound ERalpha and may contribute to tamoxifen resistance. PMID- 15351742 TI - Oligomeric Aip2p/Dld2p modifies the protein conformation of both properly folded and misfolded substrates in vitro. AB - Oligomeric actin-interacting protein 2 (Aip2p) [Nat. Struct. Biol. 2 (1995) 28]/D lactate dehydrogenase protein 2 (Dld2p) [Yeast 15 (1999) 1377, Biochem. Biophys. Res. Commun. 295 (2002) 910] exhibits the unique grapple-like structure with an ATP-dependent opening [Biochem. Biophys. Res. Commun. 320 (2004) 1271], which is required for the F-actin conformation modifying activity in vitro and in vivo [Biochem. Biophys. Res. Commun. 319 (2004) 78]. To further investigate the molecular nature of oligomeric Aip2p/Dld2p, the substrate specificity of its binding and protein conformation modifying activity was examined. In the presence of 1mM ATP or AMP-PNP, oligomeric Aip2p/Dld2p bound to all substrates so far examined, and modified the conformation of actin, DNase I, the mature form of invertase, prepro-alpha-factor, pro-alpha-factor, and mitochondrial superoxide dismutase, as determined by the trypsin susceptibility assay. Of note, the activity could modify even the conformation of pathogenic highly aggregated polypeptides, such as recombinant prion protein in beta-sheet form, alpha synuclein, and amyloid beta (1-42) in the presence of ATP. The in vivo protein conformation modifying activity, however, depends on the growth stage; the most significant substrate modification activity was observed in yeast cells at the log phase, suggesting the presence of a cofactor/s in yeast cells, where F-actin is supposed to be a major target in vivo. These data further support our previous notion that the oligomeric Aip2p/Dld2p may belong to an unusual class of molecular chaperones [Biochem. Biophys. Res. Commun. 320 (2004) 1271], which can target both properly folded and misfolded proteins in an ATP-dependent manner in vitro. PMID- 15351743 TI - Biochemical and biological responses induced by coupling of Gab1 to phosphatidylinositol 3-kinase in RET-expressing cells. AB - Grb2-associated binder-1 (Gab1) is a docking protein closely related to insulin receptor substrates. We previously reported that tyrosine 1062 in RET receptor tyrosine kinase activated by glial cell line-derived neurotrophic factor (GDNF) represents a binding site for the Shc-Grb2-Gab1 complex, and that the p85 subunit of phosphatidylinositol 3-kinase (PI3K) and SHP2 tyrosine phosphatase is associated with Gab1 in GDNF-treated cells. In the present study, we further analyzed the physiological roles of Gab1 downstream of RET, using Gab1 mutants that lack the binding sites for PI3K (Gab1 PI3K-m) or SHP-2 (Gab1 SHP2-m). Expression of Gab1 PI3K-m in SK-N-MC human primitive neuroectodermal tumor cells expressing wild-type RET markedly impaired Akt phosphorylation, Rac1 activation, and lamellipodia formation that were induced by GDNF whereas expression of Gab1 SHP2-m partially impaired Erk activation. Furthermore, expression of Gab1 PI3K-m, but not Gab1 SHP2-m, in TT human medullary thyroid carcinoma cells expressing RET with a multiple endocrine neoplasia 2A mutation enhanced cytochrome c release, and apoptosis induced by etoposide, suggesting that PI3K is involved in survival of TT cells via a mitochondrial pathway. These findings demonstrated that coupling of Gab1 to PI3K is important for biological responses in RET-expressing cells. PMID- 15351744 TI - Profiling the expression of mitogen-induced T-cell proteins by using multi membrane dot-blotting. AB - High throughput technologies are standard methods for analysis of the proteome. Multi-layer multi-well plate dot-blotting system (MLDot) technology is a high throughput dot blotting system that provides a simple, cost-effective approach for protein expression profiling in multiple samples. In contrast to traditional dot blot, MLDot uses a layered stack of thin, sieve-like membranes in place of a single nitrocellulose membrane. Therefore, up to 10 membranes can be prepared from the samples arrayed in a single 96-well plate. We describe the ability of MLDot to detect the predicted changes in protein expression following multiple mitogen treatment of T-cells. We compare the levels of the phopshorylated forms of CREB, Jun, and Akt in Jurkat T-cells as detected by MLDot to those measured by a gel-based assay. We also describe the ability of MLDot to detect differences in the levels of phosphorylated Akt in Jurkat cells as compared to primary lymphocytes. PMID- 15351745 TI - A pilot study of alcohol exposure and pharmacokinetics in women with or without children with fetal alcohol syndrome. AB - AIMS: To determine the alcohol exposure and pharmacokinetics of alcohol in a group of women who had given birth to children with FAS, compared with women who had not given birth to FAS children. METHODS: 10 women who had given birth to FAS children (FAS mothers) and 20 Controls were studied to determine how they metabolize alcohol in a single limited-access quasi-experimental session of voluntary consumption of alcohol. They had free choice in the consumption of any amount of their favourite beverage for approximately 2.5 h, but their drinking was terminated if the breath alcohol concentrations (BrAC) exceeded 150 mg%. BrACs was measured during ethanol consumption and for several hours after, for estimation of alcohol exposure and pharmacokinetics. RESULTS: FAS mothers consumed significantly larger amounts of alcohol, and achieved significantly higher peak BrAC levels than Controls. The rate of decline of alcohol from the circulation (beta-60) showed a 2-fold variation across subjects but there was no significant difference between the two groups. CONCLUSIONS: This study did not show any difference in alcohol pharmacokinetics in free-choice drinking by non pregnant women, who either have given or have never given birth to FAS children. However, mothers of FAS children tend to consume more alcohol per session. Future studies in larger samples will be needed to confirm these findings and to examine drinking patterns and other factors that may increase the risk of FAS in children of women who drink alcohol during pregnancy. PMID- 15351746 TI - Usefulness, length and content of alcohol-related discussions in primary health care: the exit poll survey. AB - AIMS: To evaluate patients' opinions of the usefulness of alcohol-related discussions with general practitioners (GPs), the time used for the discussion and its main content. METHODS: Exit poll survey to 2000 consecutive patients right after GP consultations. RESULTS: The response rate was 60.2% (1203/2000). Of the patients 11.6% (139/1203) reported that they were asked and/or advised about alcohol during the consultation. The time used for discussion about alcohol was mostly <4 min; longer for heavy than for non-heavy drinkers. Main topics of the discussion dealt with quantities consumed and harm caused by alcohol. The majority of the patients (81%) reported that discussions concerning alcohol were useful. In that respect heavy drinkers did not differ from non-heavy drinkers. CONCLUSIONS: Discussions about alcohol in primary health care were rare and short, but patients' opinions about their usefulness were mainly positive. PMID- 15351747 TI - Alcohol and injury: multi-level analysis from the emergency room collaborative alcohol analysis project (ERCAAP). AB - AIMS: To analyze the relationship between individual-level characteristics and site-level contextual variables on the association of acute alcohol use and injury. METHODS: Blood alcohol concentration (BAC) and survey data collected (using similar methodology and instruments) at the time of the emergency department (ED) visit, between 1985 and 2003 on probability samples of injured and non-injured patients (n = 18 438) from 31 EDs in seven countries (Argentina, Canada, Italy, Mexico, Poland, Spain, USA) were analyzed using hierarchical linear modeling (HLM). RESULTS: BAC and self-reported consumption were predictive of an injury (compared to a non-injury), controlling for gender and age, with odds ratios of 1.51 and 1.58, respectively. The likelihood of injury given a positive BAC and self-report was less for heavier drinkers (those reporting five or more drinks on an occasion) than for lighter drinkers, and was greater in those societies with greater detrimental drinking patterns than those with lower detrimental patterns. CONCLUSIONS: These data suggest a moderate, but robust association of a positive BAC and self-report with admission to the ED for an injury, which is modified by the patient's usual heavier drinking and by societal drinking patterns. PMID- 15351749 TI - Does group A beta-hemolytic streptococcal infection increase risk for behavioral and neuropsychiatric symptoms in children? AB - OBJECTIVE: To determine whether group A beta-hemolytic streptococcal infections increase the risk of developing symptoms characteristic of the diagnosis pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS). DESIGN: Prospective cohort study. METHODS: Children (N = 814) aged 4 to 11 years seen for sore throat or well-child care in a large pediatric practice in Rochester, NY, were enrolled from October 2001 to June 2002 (group A beta-hemolytic streptococcal [GAS] infected, n = 411; GAS uninfected, n = 403, of whom 207 had a sore throat of presumed viral etiology and 196 were well children). Symptomatic children with GAS infection (n = 399) were treated with antibiotics. At baseline and 2 and 12 weeks following baseline, all parents completed a 20-item questionnaire about the presence/absence of recent PANDAS symptoms in their children, and capable children answered 10 items about worries, obsessions, and compulsions. The relative risk of developing a "mild PANDAS variant" (> or = 2 new PANDAS symptoms) by illness type (GAS positive, presumed viral, or well child) and by parent and child report was determined and adjusted for potential covariates. RESULTS: By parent report, ill children more frequently manifested several PANDAS symptoms at baseline than well children. However, neither new symptoms nor the risk of developing a mild PANDAS variant developed during the subsequent 12 weeks more commonly in children with GAS infection than in those with presumed viral illness or in well children by parent or child report. CONCLUSIONS: Ill children with GAS infection, treated for their GAS infection, were not at increased risk for developing PANDAS symptoms or a mild PANDAS variant compared with children with presumed viral illness or well children. The role of antibiotics in the prevention or treatment of PANDAS as well as the investigation of PANDAS in the asymptomatic, infectious host deserves future research. PMID- 15351750 TI - A randomized controlled trial of an information prescription for pediatric patient education on the Internet. AB - BACKGROUND: Information prescriptions (IPs) are prescriptions of specific, evidence-based information to manage health problems. OBJECTIVE: To determine whether a pediatrician-provided IP would change parents' attitudes/behaviors about using Internet health information resources. DESIGN, SETTING, AND PARTICIPANTS: We conducted a randomized controlled trial of parents visiting an academic general pediatric practice. Both groups received a preintervention paper survey and a telephone survey 2 to 3 weeks after their clinic visit. The intervention group was offered computer training and received the IP and training summary handout. RESULTS: Final groups included 100 control and 97 intervention subjects. Of all parents, 68.0% had used the Internet in the past 6 months for health information and 52.8% used it for children's health information. The intervention group used the Internet more for general health information (P =.05) and child health information (P<.001) than the control group. At follow-up, 51 (66.2%) of 77 Internet information resources used by the intervention group were prescribed by the pediatricians. Parents who used the IP (31 [32%] of 97 intervention parents) used the Internet for general health information (P<.001) and child health information (P =.001) more than nonusers. Compared with nonusers, IP users were more likely to state they would use the IP again in the future (P =.02) and had already recommended the IP to family or friends more than nonusers (P =.001). CONCLUSIONS: Parents of children in pediatric practices commonly use the Internet for general and children's health information. In this study, IPs were associated with specific parental attitude and behavior changes resulting in increased Internet utilization for general and child health information and for specific high-quality information resources. Pediatricians can implement IPs in their office. PMID- 15351751 TI - Can an information prescription change parental attitudes and behaviors related to using the Internet for health information resources? PMID- 15351752 TI - The feasibility of universal influenza vaccination for infants and toddlers. AB - BACKGROUND: Physicians' opinions on the feasibility of routine influenza vaccination of infants and toddlers are unknown. OBJECTIVE: To assess the opinions of primary care providers regarding (1) the feasibility of an expanded influenza vaccination recommendation, (2) potential barriers, and (3) current and projected use of immunization reminder systems for influenza vaccination. METHODS: In February 2001, we mailed a 20-item, self-administered survey to a national random sample of pediatricians and family physicians (FPs). The survey primarily focused on a scenario of routine influenza vaccination for children aged 12 through 35 months using either injected or intranasal spray vaccine. RESULTS: Four hundred fifty-eight eligible physicians completed the survey (eligible response rate: pediatricians, 72%; FPs, 52%). Regarding the scenario mentioned above, most physicians agreed that implementation would be feasible (pediatricians, 80%; FPs, 69%); would significantly decrease illness visits during influenza season (pediatricians, 67%; FPs, 57%); and was justified by influenza's severity and complications (pediatricians, 61%; FPs, 41%). When considering a scenario that extended down to 6 months of age and only allowed use of injectable vaccine for infants, fewer physicians (pediatricians, 50%; FPs, 40%) considered implementation feasible. The issues most frequently cited as important potential barriers for practices were costs (77%), vaccine safety issues (52%), and the inability to identify eligible children (46%). CONCLUSION: To make widespread implementation feasible, the following are needed: minimizing costs for families and physician practices, educational campaigns on key issues, and primary care system changes (eg, tracking of eligible children, reminder and/or recall systems, and immunization clinics). PMID- 15351753 TI - A multisite randomized trial of the effects of physician education and organizational change in chronic-asthma care: health outcomes of the Pediatric Asthma Care Patient Outcomes Research Team II Study. AB - BACKGROUND: Traditional primary care practice change approaches have not led to full implementation of national asthma guidelines. OBJECTIVE: To evaluate the effectiveness of 2 asthma care improvement strategies in primary care. DESIGN: Two-year randomized controlled clinical trial. SETTING: Forty-two primary care pediatric practices affiliated with 4 managed care organizations. PARTICIPANTS: Children aged 3 to 17 years with mild to moderate persistent asthma enrolled in primary care practices affiliated with managed care organizations. INTERVENTIONS: Peer leader education consisted of training 1 physician per practice in asthma guidelines and peer teaching methods. Planned care combined the peer leader program with nurse-mediated organizational change through planned visits with assessments, care planning, and self-management support, in collaboration with physicians. Analyses compared each intervention with usual care. MAIN OUTCOME MEASURES: Annualized asthma symptom days, asthma-specific functional health status (Children's Health Survey for Asthma), and frequency of brief oral steroid courses (bursts). RESULTS: Six hundred thirty-eight children completed baseline evaluations, representing 64% of those screened and eligible. Mean +/- SD age was 9.4 +/- 3.5 years; 60% were boys. Three hundred fifty (55%) were taking controller medication. Mean +/- SD annualized asthma symptom days was 107.4 +/- 122 days. Children in the peer leader arm had 6.5 fewer symptom days per year (95% confidence interval [CI], - 16.9 to 3.6), a nonsignificant difference, but had a 36% (95% CI, 11% to 54%) lower oral steroid burst rate per year compared with children receiving usual care. Children in the planned care arm had 13.3 (95% CI, - 24.7 to -2.1) fewer symptom days annually (-12% from baseline; P =.02) and a 39% (95% CI, 11% to 58%) lower oral steroid burst rate per year relative to usual care. Both interventions showed small, statistically significant effects for 2 of 5 Children's Health Survey for Asthma scales. Planned care subjects had greater controller adherence (parent report) compared with usual care subjects (rate ratio, 1.05 [95% CI, 1.00 to 1.09]). CONCLUSIONS: Planned care (nurse mediated organizational change plus peer leader education) is an effective model for improving asthma care in the primary care setting. Peer leader education on its own may also serve as a useful model for improving asthma care, although it is less comprehensive and the treatment effect less pronounced. PMID- 15351754 TI - Prevalence and characteristics of children with special health care needs. AB - BACKGROUND: Children with special health care needs (SHCNs) are an important population from health care services, economic, and policy perspectives. However, until recently, no national data on their prevalence and health care service needs that use a commonly accepted definition have existed. OBJECTIVE: To provide national estimates of the number of children with SHCNs and their characteristics, including an assessment of how well their needs are being met. SETTING: The United States. PARTICIPANTS: Interviews were conducted by telephone with the families of 38 866 children with SHCNs younger than 18 years using the State and Local Area Integrated Telephone Survey platform developed by the Centers for Disease Control and Prevention, Atlanta, Ga. MAIN OUTCOME MEASURES: Prevalence of SHCNs, demographic and socioeconomic correlates of SHCNs, access to care, satisfaction with care, and impact on the family. RESULTS: An estimated 12.8% of US children experienced an SHCN in 2001. Prevalence was highest among boys, school-age children, and children in lower-income families. A substantial minority of these children experienced unmet health needs (17.7%) or lacked critical elements of family-centered health care (33.5%). The impact on families was pronounced, as 20.9% reported their child's health care caused financial problems, and 29.9% reported cutting back or quitting work because of their child's condition. These adverse child- and family-level impacts were concentrated among low-income and uninsured children with SHCNs. CONCLUSIONS: Children with SHCNs and their families represent an important underserved population. In addition, substantial disparities are present in access, satisfaction, and family impact. PMID- 15351755 TI - Court-mediated disputes between physicians and families over the medical care of children. AB - OBJECTIVE: To describe the judiciary's approach to parent-physician disputes over the care of sick children. DATA SOURCES: Court publications. STUDY SELECTION: Fifty parent-physician disagreements over the care of children led to physician requests for court intervention and resulted in judicial opinions published by the court. The opinions describe 66 children from 20 states. DATA SYNTHESIS: Physicians prevailed at the initial decision in 44 (88%) of the 50 disputes and at the final decision in 40 disputes (80%). Physicians were more likely to prevail in religion-based disputes than in other cases (27 of 30 vs 13 of 20; P<.03), but they were less likely to prevail in disputes concerning life threatening or potentially disabling conditions (23 of 31 vs 17 of 19; P<.19). Courts acknowledged the pediatric patients' views in only 10 of the disputes (9 of the 19 cases involving adolescents and 1 of the 31 cases involving children younger than 12 years). For most courts, the petitioning physicians provided the only source of scientific information. CONCLUSIONS: Published court opinions create precedents for future decisions and provide insight into the consequences of seeking court intervention for the physician who encounters parental refusal of care. PMID- 15351756 TI - Effectiveness of breastfeeding peer counseling in a low-income, predominantly Latina population: a randomized controlled trial. AB - BACKGROUND: Breastfeeding peer counseling has improved breastfeeding rates in developing countries; however, its impact in this country has not been adequately evaluated. OBJECTIVE: To evaluate the effectiveness of an existing, breastfeeding peer counseling program within the United States. DESIGN: Randomized, prospective, controlled trial in which participants were recruited prenatally and randomly assigned to receive either routine breastfeeding education or routine breastfeeding education plus peer counseling. SETTING: An urban hospital serving a large population of low-income Latinas. PARTICIPANTS: Pregnant women (< or =26 weeks' gestation) were recruited from the hospital's prenatal clinic. Inclusion criteria specified that participants be low income, be considering breastfeeding, have delivered a healthy, full-term singleton, and have access to a telephone. Intervention Breastfeeding peer counseling services included 1 prenatal home visit, daily perinatal visits, 3 postpartum home visits, and telephone contact as needed. MAIN OUTCOME MEASURES: Breastfeeding rates at birth and 1, 3, and 6 months postpartum. RESULTS: The proportion not initiating breastfeeding was significantly lower in the intervention group than among controls (8/90 [9%] vs 17/75 [23%]; relative risk, 0.39; 95% confidence interval, 0.18-0.86). The probability of stopping breastfeeding also tended to be lower in the intervention group at both 1 month (36% vs 49%; relative risk, 0.72; 95% confidence interval, 0.50-1.05) and 3 months (56% vs 71%; relative risk, 0.78; 95% confidence interval, 0.61-1.00). CONCLUSION: These findings demonstrate that, in the United States, peer counselors can significantly improve breastfeeding initiation rates and have an impact on breastfeeding rates at 1 and 3 months post partum. PMID- 15351757 TI - The quality of highly regarded adolescent substance abuse treatment programs: results of an in-depth national survey. AB - BACKGROUND: Drug and alcohol abuse and dependence are the most prevalent causes of adolescent morbidity and mortality in the United States. Effective, accessible treatment for adolescents with substance abuse problems is urgently needed. OBJECTIVE: To conduct the first systematic evaluation of the quality of highly regarded adolescent substance abuse treatment programs in the United States. METHODS: An advisory panel of 22 experts defined 9 key elements of effective treatment for adolescent substance abuse based on a review of the literature. In depth telephone and written surveys were conducted with 144 highly regarded adolescent substance abuse treatment programs identified by panel members and by public and private agencies. There was a 100% response rate to the initial interviews, and a 65% response rate to the follow-up surveys. The open-ended survey responses were coded by defining 5 components deemed to be crucial in addressing each of the 9 key elements, and quality scores were calculated overall and for each of the 9 key elements. RESULTS: Out of a possible total score of 45, the mean score was 23.8 and the median was 23. Top-quartile programs were not more likely to be accredited. The majority of programs scored at least 4 of a possible 5 on only 1 of the 9 key elements (qualified staff). The elements with the poorest-quality performance were assessment and treatment matching, engaging and retaining teens in treatment, gender and cultural competence, and treatment outcomes. CONCLUSIONS: Most of the highly regarded programs we surveyed are not adequately addressing the key elements of effective adolescent substance abuse treatment. Expanded use of standardized assessment instruments, improved ability to engage and retain youths, greater attention to gender and cultural competence, and greater investment in scientific evaluation of treatment outcomes are among the most critical needs. Expanding awareness of effective elements in treating adolescents will lead the way to program improvement. PMID- 15351759 TI - Impact of the Bienestar school-based diabetes mellitus prevention program on fasting capillary glucose levels: a randomized controlled trial. AB - OBJECTIVE: To evaluate the impact of a school-based diabetes mellitus prevention program on low-income fourth-grade Mexican American children. DESIGN: A randomized controlled trial with 13 intervention and 14 control schools. SETTING: Elementary schools in inner-city neighborhoods in San Antonio, Tex. PARTICIPANTS: Eighty percent of participants were Mexican American and 94% were from economically disadvantaged households. Baseline and follow-up measures were collected from 1419 (713 intervention and 706 control) and 1221 (619 intervention and 602 control) fourth-grade children, respectively. INTERVENTION: The Bienestar Health Program consists of a health class and physical education curriculum, a family program, a school cafeteria program, and an after-school health club. The objectives are to decrease dietary saturated fat intake, increase dietary fiber intake, and increase physical activity. MAIN OUTCOME MEASURES: The primary end point was fasting capillary glucose level, and the secondary end points were percentage of body fat, physical fitness level, dietary fiber intake, and dietary saturated fat intake. Fasting capillary glucose level, bioelectric impedance, modified Harvard step test, three 24-hour dietary recalls, weight, and height were collected at baseline and 8 months later. RESULTS: Children in the intervention arm attended an average of 32 Bienestar sessions. Mean fasting capillary glucose levels decreased in intervention schools and increased in control schools after adjusting for covariates (-2.24 mg/dL [0.12 mmol/L]; 95% confidence interval, -6.53 to 2.05 [-0.36 to 0.11 mmol/L]; P =.03). Fitness scores (P =.04) and dietary fiber intake (P =.009) significantly increased in intervention children and decreased in control children. Percentage of body fat (P =.56) and dietary saturated fat intake (P =.52) did not differ significantly between intervention and control children. CONCLUSION: This intervention showed some positive results, but additional research is needed to examine long-term benefits, translation, and cost-effectiveness. PMID- 15351760 TI - Characteristics associated with use or intention to use indoor tanning among adolescents. AB - BACKGROUND: Indoor tanning is a popular behavior that may increase skin cancer risk. OBJECTIVE: To examine characteristics associated with use or intention to use indoor tanning among adolescents. METHODS: A telephone interview was conducted with 1273 adolescents, aged 14 to 17 years, in the Minneapolis-St Paul, Minn, and Boston, Mass, metropolitan areas. Questions included demographic and phenotypic characteristics, knowledge, attitudes, social factors, use of indoor tanning, and intention to tan indoors. RESULTS: Twelve percent of boys and 42% of girls had tanned indoors. Among nontanners, 22.4% planned to start, and 77.2% of tanners planned to continue tanning indoors. Nontanners and tanners at risk for future indoor tanning use were each significantly more likely to be female, less likely to use sun protection, less knowledgeable about skin cancer risks, more likely to agree that tans were attractive, and more strongly influenced by social factors compared with their low-risk counterparts. CONCLUSIONS: Our data suggest that intention to tan indoors may identify a group of adolescents at risk for adopting the behavior; prospective studies are needed for confirmation. PMID- 15351761 TI - Duplicate publication. PMID- 15351762 TI - Pediatric Autoimmune Neuropsychiatric Disorders Associated With Streptococcal Infection (PANDAS): implications for clinical practice. PMID- 15351763 TI - Data sharing and duplication: is there a problem? PMID- 15351764 TI - The family (Squatting couple). PMID- 15351765 TI - Cognitive enhancement therapy for schizophrenia: effects of a 2-year randomized trial on cognition and behavior. AB - BACKGROUND: Deficits in social cognition and neurocognition are believed to underlie schizophrenia disability. Attempts at rehabilitation have had circumscribed effects on cognition, without concurrent improvement in broad aspects of behavior and adjustment. OBJECTIVE: To determine the differential effects of cognitive enhancement therapy (a recovery-phase intervention) on cognition and behavior compared with state-of-the-art enriched supportive therapy. DESIGN: A 2-year, randomized controlled trial with neuropsychological and behavioral assessments completed at baseline and at 12 and 24 months. SETTING: An outpatient research clinic housed in a medical center's comprehensive care service for patients with severe mental illness. PATIENTS: A total of 121 symptomatically stable, non-substance-abusing but cognitively disabled and chronically ill patients with schizophrenia or schizoaffective disorder. INTERVENTIONS: Cognitive enhancement therapy is a multidimensional, developmental approach that integrates computer-assisted training in neurocognition with social cognitive group exercises. Enriched supportive therapy fosters illness management through applied coping strategies and education. MAIN OUTCOME MEASURES: Six highly reliable summary measures--Processing Speed, Neurocognition, Cognitive Style, Social Cognition, Social Adjustment and Symptoms--were tested using analysis of covariance and linear trend analysis. RESULTS: At 12 months, robust cognitive enhancement therapy effects were observed on the Neurocognition and Processing Speed composites (P<.003), with marginal effects observed on the behavioral composites. By 24 months, differential cognitive enhancement therapy effects were again observed for the 2 neuropsychological composites and for Cognitive Style (P=.001), Social Cognition (P=.001), and Social Adjustment (P=.01). As expected, no differences were observed on the residual Symptoms composite. Effects were unrelated to the type of antipsychotic medication received. Enriched supportive therapy also demonstrated statistically significant within-group effect sizes, suggesting that supportive psychotherapy can also have positive, although more modest, effects on cognitive deficits. CONCLUSION: Many cognitive deficits and related behaviors of patients with stable schizophrenia are improved when sufficient exposure to relevant rehabilitation is provided. PMID- 15351766 TI - Attenuation of the neural response to sad faces in major depression by antidepressant treatment: a prospective, event-related functional magnetic resonance imaging study. AB - BACKGROUND: Depression is associated with interpersonal difficulties related to abnormalities in affective facial processing. OBJECTIVES: To map brain systems activated by sad facial affect processing in patients with depression and to identify brain functional correlates of antidepressant treatment and symptomatic response. DESIGN: Two groups underwent scanning twice using functional magnetic resonance imaging (fMRI) during an 8-week period. The event-related fMRI paradigm entailed incidental affect recognition of facial stimuli morphed to express discriminable intensities of sadness. SETTING: Participants were recruited by advertisement from the local population; depressed subjects were treated as outpatients. PATIENTS AND OTHER PARTICIPANTS: We matched 19 medication-free, acutely symptomatic patients satisfying DSM-IV criteria for unipolar major depressive disorder by age, sex, and IQ with 19 healthy volunteers. Intervention After the baseline assessment, patients received fluoxetine hydrochloride, 20 mg/d, for 8 weeks. MAIN OUTCOME MEASURES: Average activation (capacity) and differential response to variable affective intensity (dynamic range) were estimated in each fMRI time series. We used analysis of variance to identify brain regions that demonstrated a main effect of group (depressed vs healthy subjects) and a group x time interaction (attributable to antidepressant treatment). Change in brain activation associated with reduction of depressive symptoms in the patient group was identified by means of regression analysis. Permutation tests were used for inference. RESULTS: Over time, depressed subjects showed reduced capacity for activation in the left amygdala, ventral striatum, and frontoparietal cortex and a negatively correlated increase of dynamic range in the prefrontal cortex. Symptomatic improvement was associated with reduction of dynamic range in the pregenual cingulate cortex, ventral striatum, and cerebellum. CONCLUSIONS: Antidepressant treatment reduces left limbic, subcortical, and neocortical capacity for activation in depressed subjects and increases the dynamic range of the left prefrontal cortex. Changes in anterior cingulate function associated with symptomatic improvement indicate that fMRI may be a useful surrogate marker of antidepressant treatment response. PMID- 15351767 TI - The co-occurrence of DSM-IV alcohol abuse in DSM-IV alcohol dependence: results of the National Epidemiologic Survey on Alcohol and Related Conditions on heterogeneity that differ by population subgroup. AB - BACKGROUND: In DSM-IV, an alcohol abuse diagnosis is preempted by dependence, although the symptoms of each disorder are different. Consequently, little is known about the extent to which dependence occurs with or without abuse. The distinction is important because of potential heterogeneity in dependence as a phenotype in genetic research, as well as potential underestimation of alcohol dependence when surveys cover dependence symptoms only among those who screen positive for alcohol abuse. OBJECTIVE: To present the prevalence of DSM-IV alcohol dependence with and without alcohol abuse in national and population subgroups. DESIGN: Face-to-face interviews. SETTING: The United States, including Alaska, Hawaii, and the District of Columbia. PARTICIPANTS: Household and group quarters residents, 18 years and older, in the National Epidemiologic Survey on Alcohol and Related Conditions (N = 42 392). MAIN OUTCOME MEASURES: DSM-IV alcohol dependence with and without DSM-IV alcohol abuse, assessed with the Alcohol Use Disorder and Associated Disabilities Interview Schedule. RESULTS: Among respondents with current alcohol dependence, 33.7% did not additionally meet criteria for abuse (29.0% among men and 46.1% among women). Current dependence without abuse was especially common among minority women (48.5% among African Americans, 55.2% among Hispanics). Among respondents with lifetime diagnoses of dependence, 13.9% did not additionally meet criteria for abuse (10.1% among men, 22.1% among women): proportions were highest among minorities, eg, 29.1% among Hispanic women and 19.2% among Hispanic men. CONCLUSIONS: Alcohol abuse does not always accompany alcohol dependence in the general population, especially among women and minorities. Dependence with and without abuse may represent heterogeneous phenotypes for genetic research. Use of alcohol abuse as a screening method for alcohol dependence in large epidemiologic studies will differentially underestimate the prevalence of dependence by subgroup, affecting time trend and comorbidity research. Such underestimation may also perpetuate a lack of services for traditionally underserved groups. PMID- 15351768 TI - Genetic and environmental contributions to the development of alcohol dependence in male twins. AB - BACKGROUND: Information on the heritability of the development of alcohol dependence could provide a better understanding of the importance of genetic components in disease transition. OBJECTIVE: To examine the genetic and nongenetic contributions to the age at onset of regular alcohol use, the age at diagnosis of alcohol dependence, and the transition from regular alcohol use to alcohol dependence. DESIGN: Classic twin study. SETTING: General community. PARTICIPANTS: This study included 3372 twin pairs of known zygosity from the Vietnam Era Twin Registry. The diagnosis of DSM-III-R-defined alcohol dependence and related information were obtained through telephone-administered interviews conducted in 1992. MAIN OUTCOME MEASURES: Standardized proportions due to genetic vs nongenetic factors of the total variation in twin resemblance on the age at onset of regular alcohol use, the age at meeting criteria for a diagnosis of alcohol dependence, and the transition period from regular alcohol use to a diagnosis of alcohol dependence. RESULTS: Genetic influence accounted for 49% of the variation in the age at diagnosis of alcohol dependence. After adjusting for co-occurring psychiatric diseases, additive genetic factors still explained more than 37% of the variance in age at onset of alcohol dependence and at least 25% of the variance in the transition period between regular drinking and the diagnosis of alcohol dependence. Additionally, after grouping participants as early and late regular users of alcohol, the genetic effects on the transition period for early regular users were statistically significantly greater than those for late regular users. CONCLUSION: Our results demonstrate a substantial heritable basis for alcohol dependence according to its developmental sequence, including age at onset of regular use, age at diagnosis, and the transition period between regular use and diagnosis. PMID- 15351769 TI - Oral topiramate reduces the consequences of drinking and improves the quality of life of alcohol-dependent individuals: a randomized controlled trial. AB - BACKGROUND: Topiramate, a fructopyranose derivative, was superior to placebo at improving the drinking outcomes of alcohol-dependent individuals. OBJECTIVES: To determine whether topiramate, compared with placebo, improves psychosocial functioning in alcohol-dependent individuals and to discover how this improvement is related to heavy drinking behavior. DESIGN: Double-blind, randomized, controlled, 12-week clinical trial comparing topiramate vs placebo for treating alcohol dependence (1998-2001). PARTICIPANTS: One hundred fifty alcohol-dependent individuals, diagnosed using the DSM-IV. INTERVENTIONS: Seventy-five participants received topiramate (escalating dose of 25 mg/d to 300 mg/d), and 75 had placebo and weekly standardized medication compliance management. MAIN OUTCOME MEASURES: Three elements of psychosocial functioning were measured: clinical ratings of overall well-being and alcohol-dependence severity, quality of life, and harmful drinking consequences. Overall well-being and dependence severity and quality of life were analyzed as binary responses with a generalized estimating equation approach; harmful drinking consequences were analyzed as a continuous response using a mixed-effects, repeated-measures model. RESULTS: Averaged over the course of double-blind treatment, topiramate, compared with placebo, improved the odds of overall well-being (odds ratio [OR] = 2.17; 95% confidence interval [CI], 1.16 2.60; P =.01); reported abstinence and not seeking alcohol (OR = 2.63; 95% CI, 1.52-4.53; P =.001); overall life satisfaction (OR = 2.28; 95% CI, 1.21-4.29; P =.01); and reduced harmful drinking consequences (OR = -0.07; 95% CI, -0.12 to 0.02, P =.01). There was a significant shift from higher to lower drinking quartiles on percentage of heavy drinking days, which was associated with improvements on all measures of psychosocial functioning. CONCLUSIONS: As an adjunct to medication compliance enhancement treatment, topiramate (up to 300 mg/d) was superior to placebo at not only improving drinking outcomes but increasing overall well-being and quality of life and lessening dependence severity and its harmful consequences. PMID- 15351770 TI - Somatic symptoms and physiologic responses in generalized anxiety disorder and panic disorder: an ambulatory monitor study. AB - BACKGROUND: Physiologic responses of patients with anxiety disorders to everyday events are poorly understood. OBJECTIVE: To compare self-reports and physiologic recordings in patients with panic disorder (PD), patients with generalized anxiety disorder (GAD), and nonanxious controls during daily activities. DESIGN: Participants underwent four 6-hour recording sessions during daily activities while wearing an ambulatory monitor. Physiologic and subjective data were recorded every 30 minutes and during subject-signaled periods of increased anxiety or tension or panic attack. SETTING: Participants' everyday environment. PARTICIPANTS: Twenty-six patients with PD and 40 with GAD, both without substantial comorbidity, and 24 controls. INTERVENTIONS: Recordings obtained during everyday activities. MAIN OUTCOME MEASURES: Recordings of heart interbeat intervals, skin conductance levels, respirations, motion, and ratings of subjective somatic symptoms and tension or anxiety. RESULTS: Patients with anxiety disorders rated higher on psychic and somatic anxiety symptoms than did controls. Common to both anxiety disorders was diminished autonomic flexibility that manifested itself throughout the day, accompanied by less precise perception of bodily states. The main differences between patients with PD and GAD were a heightened sensitivity to body sensations and more frequent button presses. There also was a trend toward heightened basal arousal in patients with PD, manifesting itself in a faster heart rate throughout the day. CONCLUSIONS: Patients with PD or GAD are more sensitive to bodily changes than nonanxious individuals, and patients with PD are more sensitive than those with GAD. Patients with PD experience more frequent distress than those with GAD and controls, but their physiologic responses are comparable in intensity. The findings suggest that the perception of panic attacks reflects central rather than peripheral responses. The diminished autonomic flexibility observed in both anxiety conditions may result from dysfunctional information processing during heightened anxiety that fails to discriminate between anxiety-related and neutral inputs. PMID- 15351771 TI - Family transmission and heritability of externalizing disorders: a twin-family study. AB - BACKGROUND: Antisocial behavior and substance dependence disorders exact a heavy financial and human cost on society. A better understanding of the mechanisms of familial transmission for these "externalizing" disorders is necessary to better understand their etiology and to help develop intervention strategies. OBJECTIVES: To determine the extent to which the family transmission of externalizing disorders is due to a general vs a disorder-specific vulnerability and, owing to the genetically informative nature of our data, to estimate the heritable vs environmental nature of these transmission effects. DESIGN: We used structural equation modeling to simultaneously estimate the general and specific transmission effects of 4 externalizing disorders: conduct disorder, adult antisocial behavior, alcohol dependence, and drug dependence. SETTING: Participants were recruited from the community and were interviewed in a university laboratory. PARTICIPANTS: The sample consisted of 542 families participating in the Minnesota Twin Family Study. All families included 17-year old twins and their biological mother and father. MAIN OUTCOME MEASURES: Symptom counts of conduct disorder, the adult criteria for antisocial personality disorder, alcohol dependence, and drug dependence. RESULTS: Transmission of a general vulnerability to all the externalizing disorders accounted for most familial resemblance. This general vulnerability was highly heritable (h2 = 0.80). Disorder-specific vulnerabilities were also detected for conduct disorder, alcohol dependence, and drug dependence. CONCLUSIONS: The mechanism underlying the familial transmission of externalizing disorders is primarily a highly heritable general vulnerability. This general vulnerability or common risk factor should be the focus of research regarding the etiology and treatment of externalizing disorders. PMID- 15351772 TI - Prospective investigation of stress inoculation in young monkeys. AB - BACKGROUND: Retrospective studies in humans have identified characteristics that promote stress resistance, including childhood exposure to moderately stressful events (ie, stress inoculation). OBJECTIVE: Because of limited opportunities for prospective studies in children, we tested whether exposure to moderate stress early in life produces later stress resistance in a primate model. DESIGN AND MAIN OUTCOME MEASURES: Twenty squirrel monkeys were randomized to intermittent stress inoculation (IS; n = 11) or a nonstress control condition (NS; n = 9) from postnatal weeks 17 to 27. At postnatal week 35, each mother-offspring dyad underwent testing in a moderately stressful novel environment for inferential measures of offspring anxiety (ie, maternal clinging, mother-offspring interactions, object exploration, and food consumption) and stress hormone concentrations (corticotropin [ACTH] and cortisol). At postnatal week 50, after acclimation to an initially stressful wire-mesh box attached to the home cage, independent young monkeys underwent testing for inferential measures of anxiety (ie, voluntary exploration and play) in the box. RESULTS: In the novel environment test, IS compared with NS offspring demonstrated diminished anxiety as measured by decreased maternal clinging (P =.02), enhanced exploratory behavior (P =.005), and increased food consumption (P =.02). Mothers of IS offspring accommodated offspring-initiated exploration (P =.009) and served as a secure base more often compared with NS mothers (P =.047). Compared with NS offspring, IS offspring had lower basal plasma ACTH (P =.001) and cortisol (P =.001) concentrations and lower corticotropin (P =.04) and cortisol (P =.03) concentrations after stress. In the subsequent home-cage wire-box test, IS offspring demonstrated enhanced exploratory (P<.001) and play (P =.008) behaviors compared with NS offspring. CONCLUSIONS: These results provide the first prospective evidence that moderately stressful early experiences strengthen socioemotional and neuroendocrine resistance to subsequent stressors. This preclinical model offers essential opportunities to improve our understanding and enhance prevention of human stress-related psychiatric disorders by elucidating the etiology and neurobiology of stress resistance. PMID- 15351773 TI - Impact of maternal depression on infant nutritional status and illness: a cohort study. AB - BACKGROUND: The risk for emotional and behavioral problems is known to be high among children of depressed mothers, but little is known about the impact of prenatal and postnatal depression on the physical health of infants. OBJECTIVE: To determine whether maternal depression is a risk factor for malnutrition and illness in infants living in a low-income country. DESIGN: Prospective cohort study. SETTING: Rural community in Rawalpindi, Pakistan. PARTICIPANTS: Six hundred thirty-two physically healthy women were assessed in their third trimester of pregnancy to obtain at birth a cohort of 160 infants of depressed mothers and 160 infants of psychologically well mothers. MAIN OUTCOME MEASURES: All infants were weighed and measured at birth and at 2, 6, and 12 months of age, and they were monitored for episodes of diarrhea and acute respiratory infections. The mothers' mental states were reassessed at 2, 6, and 12 months. Data were collected on potential confounders of infant outcomes, such as birth weight and socioeconomic status. RESULTS: Infants of prenatally depressed mothers showed significantly more growth retardation than controls at all time points. The relative risks for being underweight (weight-for-age z score of less than -2) were 4.0 (95% confidence interval [CI], 2.1 to 7.7) at 6 months of age and 2.6 (95% CI, 1.7 to 4.1) at 12 months of age, and the relative risks for stunting (length-for-age z score of less than -2) were 4.4 (95% CI, 1.7 to 11.4) at 6 months of age and 2.5 (95% CI, 1.6 to 4.0) at 12 months of age. The relative risk for 5 or more diarrheal episodes per year was 2.4 (95% CI, 1.7 to 3.3). Chronic depression carried a greater risk for poor outcome than episodic depression. The associations remained significant after adjustment for confounders by multivariate analyses. CONCLUSIONS: Maternal depression in the prenatal and postnatal periods predicts poorer growth and higher risk of diarrhea in a community sample of infants. As depression can be identified relatively easily, it could be an important marker for a high-risk infant group. Early treatment of prenatal and postnatal depression could benefit not only the mother's mental health but also the infant's physical health and development. PMID- 15351774 TI - Genome-wide linkage analysis of a composite index of neuroticism and mood-related scales in extreme selected sibships. AB - There is considerable evidence to suggest that the genetic vulnerabilities to depression and anxiety substantially overlap and quantitatively act to alter risk to both disorders. Continuous scales can be used to index this shared liability and are a complementary approach to the use of clinical phenotypes in the genetic analysis of depression and anxiety. The aim of this study (Genetic and Environmental Nature of Emotional States in Siblings) was to identify genetic variants for the liability to depression and anxiety after the application of quantitative genetic methodology to a large community-based sample (n = 34,371), using four well-validated questionnaires of depression and anxiety. Genetic model fitting was performed on 2658 unselected sibships, which provided evidence for a single common familial factor that accounted for a substantial proportion of the genetic variances and covariances of the four scales. Using the parameter estimates from this model, a composite index of liability (G) was constructed. This index was then used to select a smaller--but statistically powerful--sample for DNA collection (757 individuals, 297 sibships). These individuals were genotyped with more than 400 microsatellite markers. After the data were checked and cleaned, linkage analysis was performed on G and the personality scale of neuroticism using the regression-based linkage program MERLIN-REGRESS. The results indicated two potential quantitative trait loci (QTL): one on chromosome 1p (LOD 2.2) around 64 cM (43-70 cM) near marker D1S2892 and another on chromosome 6p (LOD 2.7) around 47 cM (34-63 cM) near marker D6S1610. Further exploratory sex-specific analyses suggested that these QTLs might have sex limited effects. PMID- 15351775 TI - Mild overexpression of MeCP2 causes a progressive neurological disorder in mice. AB - Mutations in the X-linked methyl-CpG-binding protein 2 (MECP2), encoding a transcriptional repressor, cause Rett syndrome and a variety of related neurodevelopmental disorders. The vast majority of mutations associated with human disease are loss-of-function mutations, but precisely what aspect of MeCP2 function is responsible for these phenotypes remains unknown. We overexpressed wild-type human protein in transgenic mice using a large genomic clone containing the entire human MECP2 locus. Detailed neurobehavioral and electrophysiological studies in transgenic line MeCP2(Tg1), which expresses MeCP2 at approximately 2 fold wild-type levels, demonstrated onset of phenotypes around 10 weeks of age. Surprisingly, these mice displayed enhanced motor and contextual learning and enhanced synaptic plasticity in the hippocampus. After 20 weeks of age, however, these mice developed seizures, became hypoactive and approximately 30% of them died by 1 year of age. These data demonstrate that MeCP2 levels must be tightly regulated in vivo, and that even mild overexpression of this protein is detrimental. Furthermore, these results support the possibility that duplications or gain-of-function mutations in MECP2 might underlie some cases of X-linked delayed-onset neurobehavioral disorders. PMID- 15351776 TI - Inhibitory effect of a bitter melon extract on the P-glycoprotein activity in intestinal Caco-2 cells. AB - Extracts of bitter melon, soybean, dokudami and welsh onion by 40% methanol increased the accumulation of rhodamine-123 by Caco-2 cells, suggesting that these extracts inhibited P-glycoprotein (P-gp). The extract of bitter melon was separated in a tC18 cartridge column and the eluate from 80% acetonitrile most markedly increased the [(3)H]-daunomycin accumulation by Caco-2 cells. The inhibitory compounds in the bitter melon fraction were isolated by HPLC with Pegasil C4 and Pegasil ODS columns. The HPLC fraction having the highest activity was analyzed by (1)H-NMR and FAB-MS, and the active compound was identified as 1 monopalmitin. The inhibitory activities of 1-monopalmitin and its related compounds suggested that the inhibition of P-gp activity was not dependent on the degree of unsaturation of fatty acid in the monoglyceride, but on the chain length. It was also suggested that the monoglyceride structure played an important role in the inhibition of P-gp activity. Monoglycerides could therefore alter the pharmacokinetics of drugs by inhibiting the P-gp-mediated efflux. PMID- 15351777 TI - Nitric oxide-dependent beta2-adrenergic dilatation of rat aorta is mediated through activation of both protein kinase A and Akt. AB - Vasorelaxation to beta(2)-adrenoceptor stimulation occurs through both endothelium-dependent and endothelium-independent mechanisms, and the former is mediated through Ca(2+)-independent activation of endothelial-type nitric oxide synthase (NOS-3). Since Ca(2+)-independent NOS-3 activation may occur through its serine phosphorylation via protein kinase A (PKA) or Akt, we determined the PKA and Akt dependency of beta(2)-adrenergic relaxation of rat aorta. Rat aortic rings were pre-incubated with the PKA inhibitor H-89 (10(-7) m), the phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin (5 x 10(-7) m), Akt inhibitor (10(-5) m), or vehicle, in the absence or presence of the NOS inhibitor N(G)-nitro-l-arginine methyl ester (l-NAME, 10(-4) m). Rings were then contracted with phenylephrine (10(-7) m), and concentration-relaxation responses determined to the beta(2)-adrenoceptor agonist albuterol. Rings exhibited a concentration dependent relaxation to albuterol: pEC(50) 6.9+/-0.2, E(max) 88.2+/-4.0%. l-NAME attenuated E(max) to 60.2+/-3.5% (P<0.001). In the presence of l-NAME, wortmannin or Akt inhibitor did not influence albuterol responses, whereas H-89 reduced E(max) further, to 27.5+/-2.2% (P<0.001). In the absence of l-NAME, E(max) to albuterol was reduced by H-89, wortmannin or Akt inhibitor, to 56.2+/-2.2, 56.0+/ 1.6 and 55.4+/-1.8%, respectively (P<0.001 for each); the combinations H-89 plus wortmannin or H-89 plus Akt inhibitor reduced E(max) further still. Western blotting of NOS-3 immunoprecipitates from rat aortas confirmed that albuterol increased serine phosphorylation of NOS-3, and this increase was attenuated by H 89 or Akt inhibitor. Our results indicate that beta(2)-adrenoceptor stimulation relaxes rat aorta through both NO-dependent and independent mechanisms. The latter is predominantly PKA-mediated, whereas the former occurs through both PKA and PI3K/Akt activation. PMID- 15351778 TI - Characterization of the human HCN1 channel and its inhibition by capsazepine. AB - The human hyperpolarization-activated cyclic nucleotide-gated 1 (hHCN1) subunit was heterologously expressed in mammalian cell lines (CV-1 and CHO) and its properties investigated using whole-cell patch-clamp recordings. Activation of this recombinant channel, by membrane hyperpolarization, generated a slowly activating, noninactivating inward current. The pharmacological properties of hHCN1-mediated currents resembled those of native hyperpolarization-activated currents (I(h)), that is, blockade by Cs(+) (99% at 5 mm), ZD 7288 (98% at 100 microm) and zatebradine (92% at 10 microm). Inhibition of the hHCN1-mediated current by ZD 7288 was apparently independent of prior channel activation (i.e. non-use-dependent), whereas that induced by zatebradine was use-dependent. The VR1 receptor antagonist capsazepine inhibited hHCN1-mediated currents in a concentration-dependent (IC(50)=8 microm), reversible and apparently non-use dependent manner. This inhibitory effect of capsazepine was voltage-independent and associated with a leftward shift in the hHCN1 activation curve as well as a dramatic slowing of the kinetics of current activation. Elevation of intracellular cAMP or extracellular K(+) significantly enhanced aspects of hHCN1 currents. However, these manipulations did not significantly affect the capsazepine-induced inhibition of hHCN1. The development of structural analogues of capsazepine may yield compounds that could selectively inhibit HCN channels and prove useful for the treatment of neurological disorders where a role for HCN channels has been described. PMID- 15351779 TI - New insights into the human 5-HT4 receptor binding site: exploration of a hydrophobic pocket. AB - A body of evidences suggests that a hydrophobic pocket of the human 5-HT(4) receptor contributes to the high affinity of some bulky 5-HT(4) ligands. A thorough study of this pocket was performed using mutagenesis and molecular modeling. Ligand binding or competition studies with selected bulky ligands (RS39604, RS100235, [(3)H]GR113808 and ML11411) and small ligands (5-HT and ML10375) were carried out on wild-type and mutant receptors (W7.40A/F, Y7.43F, R3.28L) transiently transfected in COS-7 cells. The functional activity of the mutated receptors was evaluated by measuring the ability of 5-HT to stimulate adenylyl cyclase. For W7.40F mutation, no changes in the affinity of studied ligands and in the functional activity of the mutant receptor were observed, in contrary to W7.40A mutation, which abolished both binding of ligands and 5-HT induced cAMP production. Mutation R3.28L revealed a totally silent receptor with a basal level of cAMP production similar to the mock control despite its ability to product cAMP in the presence of 5-HT. Moreover, a one order loss of affinity of RS39604 and a 45-fold increase of ML11411 affinity were observed. Mutation Y7.43F modified the affinity of GR113808, which displays a 13-fold lower affinity for the mutant than for the wild-type receptor. In conclusion, in the hydrophobic pocket, two polar amino acids are able to interact through hydrogen bonds with bulky ligands depending on their chemical properties. Moreover, these experimental data may validate the proposed new three-dimensional model of the human 5-HT(4) receptor. PMID- 15351780 TI - Expression of Rho-kinase (ROCK-1 and ROCK-2) and its substantial role in the contractile activity of the sheep ureter. AB - Expression of two isoforms of Rho-kinase (ROCK) and its functional role in the physiological control of smooth muscle contraction in the sheep ureter were investigated. Helical strips of the ureteric smooth muscle were stimulated by electrical field stimulation (EFS, 60 V, 1 mS, 2, 4, 8, 16 and 32 Hz, for 20 S), KCl (80 mm), carbachol (CCh, 10(-8)-10(-4) m) or phenylephrine (Phe, 10(-8)-10( 4) m). EFS produced a reproducible contractile activity, which was abolished by tetrodotoxin (3 x 10(-6) m), a Na(+) channel blocker. A muscarinic receptor antagonist, atropine (2 x 10(-6) m), and an adrenergic neuron blocker, guanethidine (10(-5) m), significantly suppressed the contraction induced by EFS. However, this contraction was augmented in the presence of N(G)-nitro-l-arginine (l-NA, 10(-4) m), a nitric oxide synthase inhibitor. Two Rho-kinase inhibitors, Y 27632 (5 x 10(-5) m) and fasudil (5 x 10(-5) m), markedly attenuated the EFS elicited contraction. CCh and Phe produced concentration-dependent contraction in the sheep ureter. pD(2) values for Phe and CCh were 5.04+/-0.11 and 5.00+/-0.22, respectively. Y-27632 (5 x 10(-5) m) and fasudil (5 x 10(-5) m) also significantly inhibited CCh- and Phe-induced contractions. Moreover, these ROCK inhibitors produced relaxations in the KCl-elicited contraction in a concentration-dependent manner. pD(2) values for Y-27632 and fasudil were, respectively, 5.17+/-0.07 and 4.58+/-0.08 (P<0.001). Furthermore, the influences of these agents were also tested on spontaneous phasic contractions of the tissue. Among Y-27632, fasudil, TTX, l-NA, guanethidine and atropine, only the ROCK inhibitors (10(-6)-10(-5) m) were able to suppress the spontaneous contractile activity. Western blot analysis has revealed that both isoforms of Rho-kinase (ROCK-1 and ROCK-2) are expressed in the sheep ureter. Densitometric analysis has indicated that these enzymes are less expressed in the sheep ureter than are in the sheep aorta in a significant manner. These results show that a contractile enzyme, Rho-kinase, is expressed, and it mediates agonist- and EFS induced contractions as well as spontaneous contractile activity of the isolated sheep ureter. Since Y-27632 and fasudil depressed the contractions, it seems plausible to postulate that Rho-kinase inhibitors may be beneficial in the treatment of renal colic. PMID- 15351781 TI - Conformational changes in the phosphorylated C-terminal domain of rhodopsin during rhodopsin arrestin interactions. AB - Phosphorylation of activated G-protein-coupled receptors and the subsequent binding of arrestin mark major molecular events of homologous desensitization. In the visual system, interactions between arrestin and the phosphorylated rhodopsin are pivotal for proper termination of visual signals. By using high resolution proton nuclear magnetic resonance spectroscopy of the phosphorylated C terminus of rhodopsin, represented by a synthetic 7-phosphopolypeptide, we show that the arrestin-bound conformation is a well ordered helix-loop structure connected to rhodopsin via a flexible linker. In a model of the rhodopsin-arrestin complex, the phosphates point in the direction of arrestin and form a continuous negatively charged surface, which is stabilized by a number of positively charged lysine and arginine residues of arrestin. Opposite to the mostly extended structure of the unphosphorylated C-terminal domain of rhodopsin, the arrestin bound C-terminal helix is a compact domain that occupies a central position between the cytoplasmic loops and occludes the key binding sites of transducin. In conjunction with other binding sites, the helix-loop structure provides a mechanism of shielding phosphates in the center of the rhodopsin-arrestin complex and appears critical in guiding arrestin for high affinity binding with rhodopsin. PMID- 15351782 TI - T cell expression of CIITA represses Th1 immunity. AB - Despite the fact that major histocompatibility complex class II transactivator (CIITA) has been known to be involved in Th1/Th2 balance in addition to its major role as a master regulator for the expression of MHC class II genes, the exact role of CIITA in Th1/Th2 balance is still controversial. To investigate whether the Th1/Th2 balance could be modulated by T cell specific expression of CIITA, we generated CIITA-transgenic mice, in which the CIITA expression is controlled by the distal promoter of p56lck, resulting in constitutive expression of CIITA predominantly in peripheral T cells. Naive CD4+ T cells from CIITA-transgenic mice exhibited a low level of IFN-gamma secretion as well as impaired Th1 polarization in vitro, while IL-4 secretion was enhanced under Th2 condition. In addition, the development of experimental autoimmune encephalomyelitis (EAE), a prototype of Th1-mediated disease, was repressed in CIITA-transgenic mice. Resistance to EAE was correlated with reduced production of IFN-gamma in response to MOG35-55, while the proliferation of MOG35-55 -specific T cells was not affected in CIITA-transgenic mice. Together, these data demonstrate that overexpression of CIITA in T cells inhibits Th1 differentiation and function, suggesting that the expression of CIITA in T cells might play a role in the regulation of the Th1/Th2 balance during the T cell lineage commitment. PMID- 15351783 TI - Specific subsets of murine dendritic cells acquire potent T cell regulatory functions following CTLA4-mediated induction of indoleamine 2,3 dioxygenase. AB - Murine dendritic cells (DCs) expressing indoleamine 2,3 dioxygenase (IDO) catabolize tryptophan and can suppress T cell responses elicited in vivo. Here, we identify specific subsets of splenic (CD11c+) dendritic cells competent to mediate IDO-dependent T cell suppression following CTLA4-mediated ligation of B7 molecules. IDO-competent DC subsets acquired potent and dominant T cell suppressive properties as a consequence of IDO up-regulation, as they blocked the ability of T cells to respond to other stimulatory DCs in the same cultures. Soluble CTLA4 (CTLA4-Ig) and cloned CTLA4+ regulatory T cells (Tr1D1) up regulated IDO selectively in DC subsets co-expressing B220 or CD8alpha. The ability of Tr1D1 T cells to suppress CD8+ T cell responses was completely dependent on their ability to induce tryptophan catabolism in DCs. Selective IDO up-regulation in DCs did not inhibit T cell activation, but prevented T cell clonal expansion due to rapid death of activated T cells. T cell responses were restored by genetic or pharmacologic inhibition of IDO enzyme activity, or by adding excess tryptophan. DCs from interferon gamma (IFNgamma)-receptor-deficient mice were effective in promoting IDO-dependent T cell suppression following CTLA4 Ig exposure in vivo, indicating that IFNgamma signaling was not necessary for IDO up-regulation in this model. These findings suggest that IDO-competent DCs provide a regulatory bridge, mediated by CTLA4-B7 engagement, between certain regulatory T cell subsets and naive responder T cells. PMID- 15351784 TI - STAT6-mediated signaling in Th2-dependent allergic asthma: critical role for the development of eosinophilia, airway hyper-responsiveness and mucus hypersecretion, distinct from its role in Th2 differentiation. AB - When wild-type BALB/c mice were transferred with OVA-specific Th2 cells followed by OVA inhalation, a severe eosinophilia, mucus hypersecretion and airway hyper responsiveness (AHR) was induced in parallel with a marked elevation of IL-4, IL 5 and IL-13 levels in bronchoalveolar lavage fluid (BALF). However, neither eosinophilia, AHR nor mucus hypersecretion was induced in Th2 cell-transferred STAT6-/- mice. The failure of eosinophilia was not due to the defect of Th2 cytokine production in BALF of STAT6-/- mice transferred with Th2 cells, but because of the defect of STAT6-dependent eotaxin production. Indeed, intranasal administration of eotaxin reconstituted pulmonary eosinophilia but not AHR and mucus hypersecretion in OVA-inhalated STAT6-/- mice. These results initially provided direct evidence that STAT6-dependent eotaxin production is essential for pulmonary eosinophilia. We also dissociated the role of STAT6 for eosinophilia from that for AHR and mucus hypersecretion. Thus, STAT6 also plays a critical role at late phase of Th2-dependent allergy induction. PMID- 15351785 TI - Antigen-independent acquisition of MHC class II molecules by human T lymphocytes. AB - We report here that human T lymphocytes have the capacity of acquiring large amounts of MHC class II molecules from various types of antigen-presenting cells (APC) in an antigen-independent manner. The transfer of MHC class II molecules from APC to T cell required direct cell-to-cell contact and appeared to involve the interaction of numerous adhesion molecules between these cells. Depletion of cholesterol from the plasma membrane reduced the amount of MHC class II transferred onto the T cells. Most significantly, the newly acquired MHC class II molecules were capable of efficiently presenting antigen to T helper cells. These results suggest that T cells are able to interact with other T cells to regulate immune responses by presenting MHC peptide complexes that have been snatched away from nearby APC. PMID- 15351786 TI - Modulation of T cell development and activation by novel members of the Schlafen (slfn) gene family harbouring an RNA helicase-like motif. AB - The regulatory networks governing development and differentiation of hematopoietic cells are incompletely understood. Members of the Schlafen (Slfn) protein family have been implicated in the regulation of cell growth and T cell development. We have identified and chromosomally mapped four new members, slfn5, slfn8, slfn9 and slfn10, which belong to a distinct subgroup within this gene family. The characteristic feature of these proteins is the presence of sequence motifs identifying them as distinct members of the superfamily I of DNA/RNA helicases. A significant role of these newly identified members in hematopoietic cell differentiation is suggested based on their differential regulation (i) in developing and activated T cells, (ii) in LPS or IFNgamma activated macrophages, (iii) upon IL6 or LIF driven terminal differentiation of myeloblastic M1 cells into macrophage-like cells, and (iv) in splenocytes of mice infected with Listeria monocytogenes. In contrast to wild-type cells, IRF-1 and IFNalpha/betaR deficient macrophages, although undergoing growth arrest, fail to upregulate slfn gene expression upon IFNgamma or LPS stimulation, respectively. Therefore, an essential participation in IFNgamma or LPS induced growth arrest appears unlikely. Likewise, ectopic expression of the newly identified slfn family members in fibroblasts did not reveal a general impact on growth control. In contrast, transgenic T-cell specific expression of a representative member of this new subfamily, slfn8, resulted in profoundly impaired T cell development and peripheral T cells showed a reduced proliferative potential. Thus, functional participation of slfn8 in the regulatory networks governing T cell development and growth appears to be cell type specific. PMID- 15351787 TI - In vivo selection of a TCR Vbeta repertoire directed against an immunodominant influenza virus CTL epitope. AB - Little is known about the mechanisms governing TCR repertoire selection in response to foreign antigens. Here, we evaluate the molecular features of the murine C57BL/6 (B6) TCR Vbeta repertoire directed at the NP(366-374) immunodominant epitope of the influenza virus nucleoprotein. Common or 'public' beta chains are shared among individuals following either primary or secondary infection. Importantly, repertoire diversity decreases substantially after a second viral exposure due to enrichment of TCRs sharing Vbeta CDR3 loops of identical length and highly related amino acid sequences. TCRs from these secondary T cell populations possess greater overall avidity for the NP(366 374)/D(b) complex compared to those from the primary repertoire. Thus, expansion of CD8(+) T cells expressing a favored germline Vbeta gene segment in the primary response and further selection for CDR3beta loops during the secondary response, contribute to optimization of immune recognition against certain viral epitopes. PMID- 15351788 TI - The fate of autoreactive, GFP+ T cells in rat models of uveitis analyzed by intravital fluorescence microscopy and FACS. AB - Experimental autoimmune uveitis (EAU) is an inflammatory disease of the immune privileged inner eye, mediated by CD4(+) Th1 cells specific for retinal autoantigens. To elucidate the fate of the T cells in the eye we adoptively transferred green fluorescent protein-positive (GFP(+)) T cells with specificity for R14, a peptide from interphotoreceptor retinoid-binding protein (IRBP) or OVA as foreign control antigen to naive Lewis rats. We also used the model of immunogenic uveitis, an inflammatory eye disease induced by intraocular injection of soluble OVA 1 day post transfer of OVA-specific GFP(+) cells. We investigated the timing of ocular T cell infiltration and their immunological activation state by intravital fluorescence microscopy (IVFM) of the iris until onset of intraocular inflammation. Within 30 min of injection, GFP(+) cells invaded the iris tissue, irrespective of their antigen specificity, whereas intraocular inflammation was only observed 3 days later, if cells recognized their respective antigen (R14-specific cells in EAU, OVA-specific cells in immunogenic uveitis). Using FACS analysis we found that activation markers were upregulated only on cells from uveitic eyes, but not from other sources, suggesting that intraocularly presented specific antigen is a prerequisite for T cell reactivation and subsequent recruitment of inflammatory cells. PMID- 15351789 TI - Role of interstitial cells of Cajal in the control of gastric motility. AB - Most regions of the gastrointestinal tract generate spontaneous electrical and mechanical activity in the absence of stimulation. When electrical recordings are made from slow muscle cells lying in the gastrointestinal tract, a regular discharge of long lasting waves of depolarization, slow waves, is detected. It has recently become apparent that slow waves are generated by a specialized population of smooth muscle cells, known as interstitial cells of Cajal (ICC). ICC can be subdivided into at least two separate groups. In most regions of the gastrointestinal tract, one group of ICC form a network that generates pacemaker potentials, so producing rhythmical membrane potential changes in the adjacent muscle layers. The second group of ICC are distributed amongst the smooth muscle cells and are tightly electrically coupled to them. In some regions of the gut, the second group of ICC augment the waves of pacemaker depolarization, so ensuring that voltage-dependent calcium channels in the smooth muscles are activated during each slow wave cycle. In addition, the second group of ICC are densely innervated by inhibitory and excitatory nerve terminals. Thus intrinsic nerve terminals, rather than communicating directly with smooth muscle cells, selectively innervate ICC and release transmitters directly onto them. The signals that are generated in the ICC, by the neurally released transmitters, then alter the activity of surrounding smooth muscle cells. PMID- 15351790 TI - Forefront of Na+/Ca2+ exchanger studies: regulation kinetics of Na+/Ca2+ exchangers. AB - Five isoforms of Na+/Ca2+ exchanger have been identified: NCX1, NCX2, NCX3, NCX SQ1, and CALX. In all of the exchangers, the Na+/Ca2+ exchange current, which was recorded in inside-out membrane patches, was regulated by cytoplasmic Ca2+. However, the mode of regulation is different among the exchangers. NCX1, NCX2, and NCX-SQ1 are positively regulated by cytoplasmic Ca2+, but CALX is negatively regulated. NCX3 apparently has both positive and negative regulation mechanisms. In this review, I briefly summarize the Ca2+ -dependent regulation mechanisms of the exchangers. PMID- 15351791 TI - Determination of the effectiveness of components of the herbal medicine Toki Shakuyaku-San and fractions of Angelica acutiloba in improving the scopolamine induced impairment of rat's spatial cognition in eight-armed radial maze test. AB - The improving effects of various components of Toki-Shakuyaku-San (TSS) and fractions isolated from Angelica acutiloba Radix (Toki) on scopolamine-induced spatial memory impairment were investigated in eight-armed radial maze. The scopolamine-induced memory impairment was characterized by prominent increase of error choices in addition to decreased correct choices. Toki, Cnidium officinale Rhizoma (Senkyu), Poria cocos Hoelen (Bukuryo), Alisma orientale Rhizoma (Takusha), and Atractylodes lancea Rhizoma (Sojutsu) increased the correct choices, while only the Toki, Sojutsu, and Takusha decreased the error choices. No effect was produced by Paeonia lactiflora Radix (Shakuyaku). Investigation of effects of fractions isolated from Toki revealed that its activity mainly resided in the butanol layer and its contents of N-methyl-beta-carboline-3-carboxamide and amines. Moreover, the alkaloid, internal and external solutions (containing poly-, di-, and monosaccharides) obtained by dialysis with Visking cellophane tubing also improved the memory. However, no improving properties were detected for methanol and hexanol layers, L-(-)-tryptophan, L-arginine, L-(-)-lysine, and choline chloride. The results showed that the TSS components could improve the reference and working memory impaired by scopolamine. The improving effect of TSS is produced greatly by the Toki component, the activity of which was greatly produced by the fraction extracted by butanol. PMID- 15351792 TI - Pharmacological characterization of FR194921, a new potent, selective, and orally active antagonist for central adenosine A1 receptors. AB - Adenosine A1 receptors in the brain are believed to play an important role in brain functioning. We have discovered a novel adenosine A1 receptor antagonist, FR194921 (2-(1-methyl-4-piperidinyl)-6-(2-phenylpyrazolo[1,5-a]pyridin-3-yl) 3(2H)-pyridazinone), and characterized the pharmacological activity in the present study. FR194921 showed potent and selective affinity for the adenosine A1 receptor without affinity for A2A and A3 receptors and did not show any species differences in binding affinity profile among human, rat, and mouse. Pharmacokinetic study in rats revealed that FR194921 was orally active and highly brain penetrable. Oral administration of FR194921 dose-dependently ameliorated the hypolocomotion induced by the A1 receptor agonist N6-cyclopentyladenosine in rats, indicating this compound exerts A1-antagonistic action in vivo. In the passive avoidance test, scopolamine (1 mg/kg)-induced memory deficits were significantly ameliorated by FR194921 (0.32, 1 mg/kg). In two animal models of anxiety, the social interaction test and elevated plus maze, FR194921 showed specific anxiolytic activity without significantly influencing general behavior. In contrast, FR194921 did not show antidepressant activity even at a dose of 32 mg/kg in the rat forced swimming test. These results indicate that the novel, potent, and selective adenosine A1 receptor antagonist FR194921 exerts both cognitive-enhancing and anxiolytic activity, suggesting the therapeutic potential of this compound for dementia and anxiety disorders. PMID- 15351793 TI - Pharmacological characteristics of rotational behavior in hemiparkinsonian rats transplanted with mouse embryonic stem cell-derived neurons. AB - Embryonic stem (ES) cells have many of the characteristics of an optimal cell source for cell-replacement therapy. Although the usefulness of the in vitro generation of dopamine (DA)-neural precursors from ES cells has been widely discussed, functional recovery in animal models of Parkinson's disease is not fully understood. In 6-hydroxydopamine-lesioned rats, apomorphine markedly induced contralateral rotation. Apomorphine-induced rotation was significantly reduced by transplantation of neuron-like cells that had differentiated from mouse ES cells using nicotinamide, but not L-lysine. In addition, methamphetamine induced ipsilateral rotation was significantly reduced. On the other hand, picrotoxin did not inhibit apomorphine-induced rotational asymmetry. Fluoxetine alone and fenfluramine alone induced slight contralateral rotation and rotation in both directions, respectively, and these effects were similar in transplanted rats. Although immunoreactivity for tyrosine hydroxylase (TH) was almost completely lost in the ipsilateral striatum in hemiparkinsonian rats, TH immunoreactivity was detected in transplanted cells and sprouting fibers. In contrast, immunoreactivities for gamma-aminobutyric acid (GABA) and serotonin (5 HT) neurons were not changed. These results suggest that improvement of rotational behavior may be induced predominantly by transplantation of nicotinamide-treated ES cell-derived DA neurons, rather than by changes in the activities of GABA or 5-HT neural systems, in hemiparkinsonian rats. PMID- 15351794 TI - Gene expression profile in experimental mesangial proliferative glomerulonephritis. AB - We analyzed gene expression in rat anti-Thy1 antibody-induced glomerulonephritis by using the cDNA microarray method. Ninety-seven genes that differed by more than 1.5-fold intensity in comparison with the controls were selected. Cluster analysis showed that the expression of genes associated with inflammation reached maximum levels at 24 h, while genes involved in the development of fibrosis increased at 7 days after injection. Microarray analysis of animal disease models may be a powerful approach for understanding the gene expression programs that underlie these disorders. PMID- 15351795 TI - Efficacy and safety of once daily gliclazide (20 mg/day) compared with nateglinide. AB - An open-label prospective cross-over trial was performed to compare the efficacy and safety of once daily low-dose gliclazide (20 mg/day) with that of nateglinide at the usual dosage (270 mg/day, 90 mg t.i.d.) in Japanese type 2 diabetics with relatively good glycemic control (HbA1c<7.0%). Eight patients received 20 mg/day of gliclazide and 16 received 270 mg/day of nateglinide. After at least 12 weeks of gliclazide or nateglinide therapy, the drugs were switched and treatment was continued for another 12 weeks. The final HbA1c value was modestly, but significantly, lower after gliclazide treatment than after nateglinide treatment (6.2% vs. 6.4%). However, symptoms related to hypoglycemia were significantly more common with gliclazide treatment than nateglinide treatment (7 vs. 0 cases), although there were no severe hypoglycemic events. While gliclazide acts as a free radical scavenger, there was no effect on parameters of oxidative stress such as malondialdehyde-modified low density lipoprotein and thiobarbituric acid reactive substances at the low dosage tested. In conclusion, both drugs are reasonable options for early type 2 diabetes. Compared with the regular dose of nateglinide, 20 mg/day of gliclazide achieved modestly better glycemic control with an increased frequency of hypoglycemia in diabetic patients with relatively good glycemic control. PMID- 15351796 TI - Immunohistochemical analysis of bcl-2, Bax and Bak expression in thyroid glands from patients with Graves' disease. AB - In order to clarify the role of apoptosis and the expression of Bcl-2 family proteins in the pathology of Graves' disease (GD), we evaluated the apoptosis by in situ end-labeling of fragmented DNA and the expression of Bcl-2, Bax and Bak by immunohistochemistry in thyroid tissues from 20 patients with GD and in normal thyroid tissues from 6 patients with follicular adenoma (N). Apoptotic nuclei were found in thyrocytes and in germinal center of lymphoid follicles. Bcl-2 was strongly expressed in both GD and N thyrocytes. Bax was not expressed in either GD or N thyrocytes. Bak was expressed in thyrocytes from 5 of 20 patients with GD, while it was detected in all N thyrocytes. In lymphoid follicles Bcl-2 was expressed in the mantle zone, while Bax and Bak were both expressed in the germinal center. The percentage of apoptotic nuclei in GD thyrocytes was low (0~3.6%), and negatively correlated with the weight of the thyroid glands resected (rs = -0.43, P<0.05). It was greater in Bak-positive GD thyrocytes than in Bak-negative ones (mean +/- SD; 1.7 +/- 0.7% vs. 0.7 +/- 0.9%, P<0.05). These findings suggest that the differential expression of Bcl-2 family proteins in both thyrocytes and lymphoid follicles may be involved in the pathology of GD. PMID- 15351797 TI - Lessons learnt from influences of the Marmara earthquake on glycemic control and quality of life in people with type 1 diabetes. AB - To examine the short- and long-term influences of the Marmara earthquake, which occurred on August 17, 1999 in Turkey, on glycemic control and quality of life (QOL), HbA1c, insulin requirement and QOL of 88 people with type 1 diabetes living in the quake zone were evaluated one year before (PreE), 3 months after (PostE) and one year after (FE) the earthquake. HbA1c levels and daily insulin requirements increased significantly at PostE (HbA1c from 7.4 +/- 1.3% to 8.5 +/- 1.8%, p<0.05; insulin from 0.58 +/- 0.2 IU/kg/day to 0.77 +/- 0.2 IU/kg/day, p<0.05). Mean total QOL scores at PostE were significantly lower than the scores obtained at PreE (62.7 +/- 17.3 vs 74.2 +/- 13.4, p<0.001). There were no significant differences between HbA1c levels and total QOL scores at PreE and FE. People with type 1 diabetes living in the same house after the earthquake and not having enough food supply were reported to have lower QOL than people moving to another house and having enough food supply after the earthquake (p = 0.014, p<0.0001, respectively). The Marmara Earthquake had a negative impact on the glycemic control and QOL of the subjects with type 1 diabetes for the short term but prequake scores might be achieved after a long period. PMID- 15351798 TI - Leptin regulation of the thyroids: negative regulation on thyroid hormone levels in euthyroid subjects and inhibitory effects on iodide uptake and Na+/I- symporter mRNA expression in rat FRTL-5 cells. AB - Leptin receptors are distributed throughout the body and leptin has been shown to have various effects. As we have recently demonstrated a positive correlation between serum leptin levels and TSH in euthyroid subjects, we investigated the effect of leptin on the thyroids. It was observed that serum leptin levels were negatively correlated with free thyroxine/TSH ratios in the serum of euthyroid female subjects. This suggests that leptin may modulate TSH effects. RT-PCR for leptin receptor expression revealed that FRTL-5 cells possess the gene transcript to the long cytoplasmic form of the receptor. Leptin actually appeared to induce an increase in c-fos mRNA expression. However, it inhibited iodide uptake typically induced by both TSH and dibutyryl cAMP, while leptin did not inhibit TSH-induced cAMP production or TSH-stimulated DNA synthesis in 4H medium (in the absence of insulin and TSH). Leptin also was observed to inhibit TSH- and dibutyryl cAMP-induced Na+/I- symporter and thyroglobulin mRNA expression. Lastly, leptin was seen to inhibit TSH-stimulated thymidine incorporation in 5H medium. Taken together, these results suggest that leptin suppresses TSH-induced thyroid function. Therefore, we hypothesized that leptin may be one of the regulators of thyroid function in obese patients. PMID- 15351799 TI - Endocrinological and pathological effects of anabolic-androgenic steroid in male rats. AB - Many athletes use drugs, especially anabolic androgenic steroids (AAS), but there are few reports on the endocrinological and pathological changes in AAS abusers. In this study we reported the results of endocrinological examinations in rats administered AAS and also physical changes. We separated 37 male Wistar rats (7 weeks old) into 3 groups: Group A was medicated with nandrolone decanoate, metenolone acetate, and dromostanolone; Group B with nandrolone decanoate and saline; and Group C was given only saline. They were given subcutaneous injections of the medications or the control vehicle once a week for 6 weeks. Medications were stopped for 4 weeks, and then resumed for another 6 weeks. After that, rats were sacrificed. Serum testosterone level in Group A was significantly higher than that in Group C. Serum dihydrotestosterone in Group A was significantly higher than that in both Groups B and C. Serum estradiol-17beta levels in Groups A and B were significantly higher than that in Group C. In pathological evaluation, heart, testis, and adrenal gland were severely damaged. These findings indicate that there is a high degree of risk related to the use of AAS. PMID- 15351800 TI - SIADH closely associated with non-functioning pituitary adenoma. AB - We demonstrated severe hyponatremia in a 68 year-old man who had pituitary tumor. He had poor appetite and was disoriented. Tests revealed hyponatremia of 110 mmol/l, and he was admitted to Jichi Medical School Omiya Medical Center to undergo further tests. Physical findings revealed disturbance of consciousness with Japan Coma Scale I-2. There was neither dehydration nor edema. Laboratory data showed a serum sodium level of 112 mmol/l; plasma osmolality, 219 mmol/kg; and urinary osmolality, 555 mmol/kg. Plasma arginine vasopressin (AVP) level was 1.6 pmol/l despite the marked hypoosmolality. Anterior pituitary function was normal. Brain magnetic resonance imaging showed a pituitary tumor of 20 x 18 x 20 mm in size, which pushed the pituitary stalk upward. After the adenomectomy, serum sodium level was kept normal without any treatment. Histology showed basophilic adenoma. These findings indicate that local pituitary tumor may cause exaggerated secretion of AVP, resulting in the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). PMID- 15351801 TI - Appearance of thyroid stimulating and blocking immunoglobulins after bone marrow transplantation: presentation of two contrasting cases. AB - Two acute leukemia cases who presented autoimmune thyroid diseases after bone marrow transplantation (BMT) are described with reference to the pathogenesis of their autoimmune clones. A 37-year old Japanese woman developed Graves' hyperthyroidism 39 months after allogeneic BMT for acute myeloid leukemia (AML) donated from her sister. Although both donor and recipient were euthyroid and negative for thyroid autoimmunity before BMT, the donor was positive for anti nuclear and anti-single strand DNA autoantibodies. Studies on polymorphism for variable number of tandem repeat region of T-cell receptor gene suggested that the lymphocytes responsible for the hyperthyroidism were of donor origin. The second case was a 12-year-old Japanese schoolboy who presented nongoitrous hypothyroidism 2 years after autologous BMT for acute lymphoblastic leukemia (ALL). He had been clinically euthyroid before transplantation. Family history revealed that his mother and sister had a history of Graves' disease. His serum was positive for thyroid-stimulation blocking antibody. It is highly likely that the autoimmune process was activated after transient immune suppression during peri-BMT period in this patient. Pathogenesis, incidence, and observed time lag between BMT and development of autoimmune thyroid diseases were discussed. PMID- 15351802 TI - Prostatic adenocarcinoma metastasis in the thyroid gland. AB - Metastasis from prostate to thyroid gland is very uncommon. Here we report a 77 year-old man who was admitted to the hospital because of a nodular goiter. A fine needle aspiration biopsy of the nodule showed metastatic prostatic adenocarcinoma. This is the second case of a metastatic prostate carcinoma to the thyroid gland. PMID- 15351803 TI - [Health economics analysis of colorectal screening]. AB - AIM: To assess the screening rate, the cost of screening and treatment, and to calculate the expected epidemiological and economic gain and cost-effectiveness of mass-screening programme. METHODS: The data derive from the financial database of the National Health Insurance Fund of Hungary from 2001. The cost of treatment includes the cost of outpatient care, the acute and chronic inpatient care, the subsidies of medicines' prices and the expenditure on disability to work (including sickness-pay). The expected benefits of the screening programme were modeled with different screening strategy and mortality decrease for a 10 years interval. RESULTS: The cost of treatment of colorectal cancer was around 9.98 billion Hungarian forints (34 817 250 USD, 38 871 666 EUR) in 2001. In the age group 45-65 with 10% mortality decline 718 lives (net present value, NPV: 515), with 20% mortality decline 1462 (NPV: 1050) lives can be saved during a 10 years screening programme. The cost of one life saved varies between 4.0 million Hungarian forints (13968 USD, 15595 EUR)/life saved and 16.3 million Hungarian forints (56.952 USD, 63.584 EURO)/life saved according to the mortality decline and screening strategy. The cost of one life year saved varies between 307 909 Hungarian forints (1074 USD, 1200 EUR)/life year saved and 1.25 million Hungarian forints (4381 USD, 4891 EUR)/life years saved. CONCLUSION: The implementation of organized colorectal screening can lead to cost saving in Hungary. The cost effectiveness of colorectal screening seems to be acceptable for purchaser, but many methodological and organizational issues should be discussed in details. PMID- 15351804 TI - [Predictive value of human genotoxicological biomarkers in the primary prevention of chronic non-infectious diseases]. AB - The main goal of biomarker research in the frame of primary prevention of chronic diseases is the prevention of appearance of clinical symptoms by an early recognition of the process leading to the symptoms. By the use of well established biomarkers one can detect such tendencies finally leading to the manifestation of the disease far before the progress turns irreversible. As several parameters play role in such processes, the estimation of biological changes with the help of biomarkers is a precise and relatively simple means of the prevention. Indications of intervention in order to prevent the manifestation of a disease are rather determined by the nature but not the number of alterations. The use of genetic screening and monitoring by adequate biomarkers provide us with a new opportunity to measure such qualitative changes rather than the quantitative ones, ensuring a great improvement to the previous methods. The use of qualitative parameters as a routine method instead of the classic quantitative measures might represent a challenge for current prevention policies in Europe to approach health problems and safety at work. When the biomarkers give positive results that means a high probability to be at risk but not yet the manifestation of a certain illness. This stage may be relevant to the early onset of certain pathological processes, but it does not necessarily turn to a performed disease. In consequence a so-called positive result might cause unnecessary disturbances for the probands leading to ethical issues of risk communication. The monitoring system, however, should find acceptable communication strategies for the high-risk conditions detected by the well established biomarkers. PMID- 15351805 TI - [Cellular biology and public health]. AB - Within the framework of the National Research and Development Program No. 1/016 /2001 the authors determined the population levels of ionizing and nonionizing radiations, elaborated the system of assays for detection of biological effects including dose-effect relationships, studied the roles of protective and sensitizing factors in the effect modification. The radium content of building materials were determined as well as the indoor radon activity concentrations and the magnetic induction fields around household equipment operating with 50 Hz. Biological dosimetry techniques were categorized according to the indication time. In addition, radiation sensitivity of intracellular antioxidant enzymes and the antioxidant capacity of blood sera were measured. PMID- 15351806 TI - [Biomarker indicators of chemoprevention among subjects occupationally exposed to metals]. AB - Chemoprevention with chelating agent Humetta for three months was performed, due to anaemia and other haematologic disorders, immunotoxicological alterations and/or increased chromosome aberration rate among galvanisers and goldsmiths occupationally exposed to precious and heavy metals. Twenty-two of altogether 47 subjects took part voluntarily in the chemoprevention, and the rest of the subjects served as untreated controls. Complex clinical laboratory testing including detailed anamneses; genotoxicological and immunotoxicological monitoring were performed before and after administration of chemopreventive agent. After chemoprevention a significant improvement was observed in anaemia and serum glucose levels, while a less marked improvement was found in serum cholesterol levels and liver functions. Altered chromosome aberration and apoptotic cell fraction also tended to normalise after treatment. Immunological parameters were not affected by the treatment. The obtained results may suggest that chemoprevention with chelating agents as Humetta can help in the prevention of harmful effects of occupational exposures to metals. PMID- 15351807 TI - [Factors influencing lycopene content of foods, and lycopene intake of Hungarian population]. AB - Lycopene is an acyclic, biologically active carotenoid that constitutes foods, its preventive role in several cancerous diseases have been proved by epidemiological and experimental data. Its beneficial role in maintenance of human health is related to its significant antioxidant properties. Data of dietary lycopene intake of the Hungarian population is not available. The aims of the present complex study were 1) to measure the lycopene content of foods frequently consumed in Hungary, 2) to investigate the effect of agrotechnological procedures and food processing on lycopene content of tomatoes, 3) to estimate the lycopene intake in two groups of the Hungarian population with the use of a three-day dietary record. The best lycopene sources are the raw (5.0-16.0 mg/100 g) and processed tomatoes and tomato products (3.0-80.0 mg/100 g), and also watermelon (3.6-6.2 mg/100 g). The variety of the plants, the growing circumstances, and the weather conditions significantly influence the lycopene content of freshly consumed and processed tomato fruits. Mild technological processes can preserve a considerable amount of the original lycopene content in tomato. The estimated average dietary intakes of the Hungarian children (n=521) and adults (n=205) were 2.98 +/- 4.71 mg/day/capita, and 4.24 +/- 8.47 mg/day/capita, respectively. Optimal climate conditions of Hungary makes possible to produce tomato fruits with high dietary value including significant amount of health protective lycopene. Increased consumption of tomato and tomato products with high concentration of lycopene may improve the antioxidant capacity of human body, and the risk of several cancerous diseases may be reduced. PMID- 15351808 TI - [Characterization of chemically exposed groups by immunotoxicological methods]. AB - At the National Institute of Chemical Safety we have surveyed the immunological status of donors from the oil industry, health services, and metallurgy exposed to different immunotoxic compounds. Their data were compared to those of healthy, non-exposed controls. Our aim was to study the relationship between immunotoxic exposure and immune function, and to establish a system of immunological parameters by which chemical exposure can be specifically monitored. Subpopulations and activation of lymphocytes was measured by flow cytometry, using immunophenotyping of peripheral blood lymphocytes. In the groups exposed to immunotoxic compounds we found an increase in helper, and a decrease in cytotoxic T lymphocytes, leading to a shift in Th/Tc ratios. These phenomena are not substance specific, but relate to chemical exposure. The lymphocytes of exposed groups showed a higher proportion of activated cells, but there was a difference in the expressed activation markers. Our results suggest that characterizing lymphocyte subpopulations and activation markers on PBL of donors is a useful tool in tracking environmental immunotoxic effects. PMID- 15351809 TI - [Evaluation of high-dose-rate brachytherapy of the preoperative treatment of stage IB (FIGO) cervical carcinoma]. AB - PURPOSE: Examination of stage IB (FIGO) cervical cancer patients treated with preoperative high dose rate brachytherapy (HDR BT) on the basis of specimen histology, local control and disease-free survival. MATERIAL AND METHOD: Patients with stage IB cervical cancer were treated between 01/01/1993 and 31/1/1997 as part of complex treatment with HDR BT, the dose calculated for point A 2x5.5 Gy, with one week interval in the Municipal Centre of Oncoradiology, Uzsoki Hospital, Budapest. Different types of surgery were carried out in general within two weeks after preoperative HDR BT. Depending on the specimen histology, postoperative radiation treatment varied. In case of residual tumor vaginal BT 2x5.5 Gy was applied within a week; the dose was relevant to a tissue depth of 5 mm and to the upper third of vagina. This was followed by percutaneous irradiation (telecobalt, linear accelerator) at a dose of 50 Gy, box technique. In certain cases the para aortal region was simultaneously irradiated. In case of negative specimen no postoperative vaginal HDR BT was applied in the majority of patients. Retrospective analysis helped to determine the effectiveness of regularly followed patients, as for local control and metastasis on the basis of the time of development of dissemination, respectively. RESULT: 33.4% of 153 preoperatively treated patients had tumor-free specimen. Regular follow-up (at least 5 years) showed local relapse in 17 patients (11%), dissemination in 13 patients (8.4%), and 4 among them (2.6%) had simultaneous local relapse. CONCLUSION: On the basis of our data it can be stated that preoperative HDR BT in stage IB cervical carcinoma decreased local relapse. In spite of negative specimen histology, vaginal HDR BT can be suggested, but if there is residual tumor, both HDR BT and percutaneous irradiation is suggested on the basis of our data. PMID- 15351810 TI - [Radiotherapy in combination with low-dose chemotherapy in locally advanced head and neck cancer]. AB - A previous preclinical study revealed that the maximal additive effect between chemotherapy (CT) and irradiation (RT) occurred at a low level of CT. Therapy was therefore designed with an oral drug daily given in combination with RT in order to determine the efficacy and toxicity. Locoregionally advanced head and neck tumor patients were treated with simultaneous RT and CT. RT was administered 5 times per week at 2 Gy per fraction in a total dose of 70 Gy. Throughout the treatment 30 mg/kg Tegafur was given daily orally. In the period between 2000 and 2002, 50 patients were enrolled. Complete remission was attained in 60%, with an overall response rate of 94%. Acute mucositis of grade 2 or 3 was observed in 56% (28 patients), and gastrointestinal and hematologic toxicity of grade 2 or 3 occurred in 8% (4 patients). Because of side-effects, the duration of treatment was at most 2 weeks longer. Toxicity was eliminated quickly by careful supportive therapy. In conclusion, it is considered that oral low-dose CT in combination with RT is an efficient and simple mode of treatment for locally advanced head and neck tumor patients with a poor prognosis. PMID- 15351811 TI - [The radiochemotherapy of the oesophageal cancer]. AB - AIM: To prove the efficacy of the simultaneous radio-chemotherapy of the inoperable or non-resectable esophageal cancer. METHODS: Twenty-nine patients with inoperable or non-resectable oesophageal cancer were treated between January 1995 and December 2002. The therapy was started with intraluminal HDR AL irradiation for the recanalisation of the esophagus (8 Gy at 0.5 cm depth, repeated two-three times, with one-week interval), followed by percutaneous megavolt irradiation one week after the last HDR AL session (50 Gy total dose, 5 x 2 Gy/week fractions for 5 weeks). The chemotherapy was started simultaneously with the percutaneous megavolt irradiation (three courses of Cisplatin-5 Fluorouracil combination, with four-week intervals). RESULTS: The swallow function has been improved in 16/29 patients, it remained unchanged in 10/29 and got worse in 3/29 patients (1 and 3 Units), respectively. REMISSION: complete 9/29 patients, partial 17/29 patients. Side effects: Esophagitis of different degree occurred in all patients, consecutive transitory dysphagia developed in 8/29 patients, leucopenia after the chemotherapy in 2/29 patients, tracheo esophageal fistula in 3/29 patients. Follow-up time: average 12.2 months (3-55 months). The duration of the swallow function improvement: average 10.7 months (2 55 months). CONCLUSION: The initial results refer to the favourable effect of the palliative radio-chemotherapy of the inoperable esophageal cancer. PMID- 15351812 TI - [Treating Hodgkin's disease: for whom the more means better?]. AB - During the last half century, our knowledge and expertise increased, and Hodgkin's disease became one of the most curable malignant diseases. However, our results are still not satisfactory in the case of advanced stage. Apart from standard chemotherapy regimens, several other, so-called intensive first line therapy schedules have been tried, whose common characteristic is that more antineoplastic drugs within a reasonably shorter period and sometimes with increased dose had been administered. Out of these regimens, escalated dose BEACOPP (bleomycin, etoposide, adriamycin, cyclophosphamide, vincristine, procarbazine, prednisolone), the therapy yielding the best results and inciting hot debates in the literature is discussed here in detail. Though many questions related to BEACOPP scheme are still unanswered and at present it is not recommended for use in first line therapy except for trials, it seems that patients under 60 years of age, with advanced stage disease having poor prognosis, may benefit from this treatment. PMID- 15351813 TI - [Pure red-cell anemia due to recombinant human erythropoietin and its status in oncology practice]. AB - A special form of anemia was observed during the treatment with recombinant human erythropoietin having been applied for more than 15 years. The pure red-cell anemia due to antibodies against erythropoietin was described in chronic renal failure patients. Since oncological patients are also treated with rhuEPO it is interesting to know whether this side effect could be observed in the patients with solid tumors as well. It should be considered in tumorous patients when coexistence of antibodies against rhuEPO with pure red-cell aplasia is demonstrated, and its other causes as immunological disease, thymoma, viral infections (eg. Parvovirus B12, Hepatitis B or C) are excluded. The author collected literature data and found the absence of reports on this side effect in cases of treatment with rhuEPO in cancer patients. The rhuEPO treatment is a safe method for the cure of cancer anemia as antibodies against rhuEPO have not been shown together with PRCA among cancer patients. The possible explanation could be the shorter application time in cancer compared to the chronic renal failure patients. The side effect observed in chronic renal failure patients calls the attention to the precise compliance with the instructions of the manufacturers. PMID- 15351814 TI - [Diagnostic imaging of cancer]. AB - Rapid development of imaging techniques provided much more precise methodology of diagnosis, staging and dynamics of cancer. Nowadays the onco-radiodiagnostic units are able to select the optimal imaging technique based on established international protocols. These protocols provide the basis of cancer diagnosis, therapy control and clinical research. The onco-radiodiagnostic unit is essential part of the oncoteam managing the cancer patients. Follow-up protocols are now equally important compared to those of the diagnosis and staging, requiring a continuous interaction between radiologists and physicians. The comprehensive cancer centers with all the necessary imaging techniques are the optimal organizations where professional and economic priorities both can be considered for the benefit of cancer patients. PMID- 15351815 TI - The renaissance of palladium(II)-catalyzed oxidation chemistry. AB - Palladium(II)-catalyzed oxidations constitute a paramount reaction class but have remained immature over the past few decades. Recently, this field has reappeared at the forefront of organometallic catalysis. This emerging area article outlines recent developments in palladium(II)-catalyzed oxidation chemistry with discussion of potential future growth. PMID- 15351816 TI - Cross-metathesis coupling of sugars and fatty acids to lysine and cysteine. AB - Attachment of an olefin tether to the side chain of either lysine or cysteine allows cross metathesis (CM) conjugation with olefin-containing sugar and fatty acid analogues. PMID- 15351817 TI - Regio- and stereoselective hydrosilylation of terminal alkynes using Grubbs' first-generation olefin-metathesis catalyst. AB - Grubbs' first-generation, Ru metathesis complex catalyses the hydrosilylation of terminal alkynes. The reaction exhibits an interesting selectivity profile that is dependent on the reaction concentration and more importantly on the silane employed. PMID- 15351818 TI - Atrazine transformation using synthetic enzymes prepared by molecular imprinting. AB - Atrazine imprinted synthetic polymers were prepared using a combination of methacrylic acid and 2-sulfoethyl methacrylate that bound and converted atrazine and other 6-chlorotriazine herbicides to less toxic compounds. A chloride at the 6-position of the triazines was converted to a methoxy group by the polymer in a methanol-containing solvent, where the imprinting effects enhanced the catalytic activity. Competitive inhibition of the atrazine methanolysis was observed in the presence of a structurally related binder, ametryn, suggesting that the catalytic reaction proceeded in the binding sites generated by the molecular imprinting process. PMID- 15351819 TI - The Strecker reaction: kinetic and equilibrium studies of cyanide addition to iminium ions. AB - Kinetics studies are reported of the reactions of benzylidene benzylamine 4a, and of benzylidene allylamine 4b, with cyanide in aqueous buffers to give the corresponding [small alpha]-aminonitriles. The results allow the calculation of values of rate and equilibrium constants for reaction of the iminium ions formed from 4a and 4b with cyanide ions. These values are compared with those, obtained from the hydrolysis reactions, for reaction of the iminium ions with hydroxide ions and with water. Comparison with some other iminium ions reveals that those formed from 4a and 4b are relatively unreactive due to the possibilities of charge delocalisation. PMID- 15351820 TI - Is the ring conformation the most critical parameter in lipase-catalysed acylation of cycloalkanols? AB - CAL-B catalysed the resolution of several five and six-membered cyclic beta hydroxy esters efficiently with the exception of the cis-cyclohexanol (+/-)-4. When employing molecular modelling techniques the conformation turned out to be the most important determinant for their reactivity towards O-acetylation. In all cases, the R enantiomers reacted faster than the S enantiomers since the reactive intermediates of the former can adopt more favourable ring conformations and thus experience less steric hindrance in the active site. Furthermore, the minimised structure for the main conformer of R-4 showed that the axial hydrogens in the 3 and 5-positions with respect to the hydroxyl group prevent the enzymatic reaction. PMID- 15351821 TI - Reactivity of hydrazinophthalazine drugs with the lipid peroxidation products acrolein and crotonaldehyde. AB - The nucleophilic drug hydralazine strongly inhibits cell toxicity mediated by acrolein, a short chain 2-alkenal formed during lipid peroxidation. We here report the chemistry of acrolein-trapping by hydralazine, and show that together with its structural analogue dihydralazine, it also readily traps crotonaldehyde. Isolable reaction products included (1E)-acrylaldehyde phthalazin-1-ylhydrazone (E-APH), (1Z)-acrylaldehyde phthalazin-1-ylhydrazone (Z-APH), (1E,2E)-but-2-enal phthalazin-1-ylhydrazone (E-BPH) and (1Z,2E)-but-2-enal phthalazin-1-ylhydrazone (Z-BPH). Concentration-dependent formation of (1E)-acrylaldehyde phthalazin-1 ylhydrazone was observed in the culture media of cells co-exposed to hydralazine and the acrolein precursor allyl alcohol. These aldehyde-sequestering properties of hydrazinophthalazine drugs may contribute to the protection they provide against 2-alkenal-mediated toxicity. PMID- 15351822 TI - Rigid versus flexible: how important is ligand "preorganization" for metal ion recognition by lower rim-functionalized calix[4]arenes? AB - For an assessment of the outcomes from use of an appropriately "preorganized" calixarene-based ionophore versus its conformationally mobile prototype, solvent extraction propensities of flexible calix[4]arene di-[N-(X-sulfonyl)carboxamides] for alkali, alkaline earth metal cations, Pb2+, Ag+ and Hg2+ are compared with those for seven new rigid analogs fixed in the cone, partial cone and 1,3 alternate conformations. For each of the metal ions, the preferred calix[4]arene conformation was determined from the NMR spectra for the metal salt of the flexible ligand. Except for Ag+, flexible calix[4]arene di-[N-(X sulfonyl)carboxamides] were found to provide greater metal ion extraction efficiency and better selectivity than the corresponding "preorganized" ionophores. PMID- 15351823 TI - Conjugation of a novel histidine derivative to biomolecules and labelling with [99mTc(OH2)3(CO)3]+. AB - The new histidine derivative 3-[1-[3-(9H-fluoren-9-ylmethoxycarbonylamino) propyl]-1H-imidazol-4-yl]-2-(3-trimethylsilanyl-ethylcarboxyamino)-propionic acid methyl ester (7) has been prepared via alkylation of the histidine urea derivative (7S)-5,6,7,8-tetrahydro-7-(methoxycarbonyl)-5-oxoimidazo-[1,5-c] pyrimidine (2) with Fmoc-protected 3-iodopropyl-amine, followed by ring opening with 2-trimethylsilylethanol. After Fmoc cleavage by HNEt2, the histidine amine derivative was coupled to biotin, to the pentapeptide leucine-enkephalin and to Vitamin B12-b-acid by amide formation, employing TBTU as the coupling reagent. In order to make the histidine accessible for labelling, the teoc protecting group was removed by either NBu4F (for the biotin conjugate) or by TFA (for the enkephalin and B12 conjugates). Reaction of a 10(-4) M solution of the bioconjugates with [99mTc(H2O)3(CO)3]+ at 50 degrees C for 30 min led to the formation of one single new peak in the HPLC radiochromatogram in each case, confirming quantitative labelling of the respective biomolecules. To assess the nature of the labelled compounds, the rhenium analogues with Re(CO)3 were also synthesised and similar retention times confirmed the identity with the 99mTc labelled conjugates. PMID- 15351824 TI - Chimeric (aeg-pyrrolidine)PNAs: synthesis and stereo-discriminative duplex binding with DNA/RNA. AB - The design and facile conversion of naturally occurring 4-hydroxyproline to all four diastereomers of thymine pyrrolidine PNA monomer, (2R,4S)-adenine, -guanine and -cytosine monomers and their incorporation into duplex forming PNA oligomers is reported. The interesting results of the hybridization studies with complementary DNA/RNA sequences in either parallel or antiparallel orientation reveal the stereochemistry-dependent DNA vs. RNA discriminations and parallel/antiparallel orientation selectivity. PMID- 15351825 TI - Preparation of novel selenapenams and selenacephems by nucleophilic and radical chemistry involving benzyl selenides. AB - 2,2a-Dihydro-1H,8H-azeto[2,1-b][1,3]benzoselenazin-1-one (12), 5-selena-1 azabicyclo[4.2.0]oct-3-en-8-one (13), ethyl 1-aza-7-oxo-4 selenabicyclo[3.2.0]heptane-2-carboxylate (16), and benzoselenopenem (33) can be prepared in 39-85% yield through the intramolecular homolytic substitution of aryl, vinyl or alkyl radicals at the selenium atom in suitably-substituted 4 benzylseleno-beta-lactams, or through intramolecular nucleophilic substitution by the benzylseleno moiety in 4-halo-beta-lactam precursors. Application of this chemistry to the preparation of optically active selenium-containing analogues of beta-lactam antibiotics is also detailed. PMID- 15351826 TI - Supramolecular ion channels from a transmembrane bischolic acid derivative showing two discrete conductances. AB - Bischolic acid derivative 1 linked by m-xylylene dicarbamate at the 3,3'-position was synthesized and the single ion channel properties were examined. Compound 1 showed two clearly distinct conductances, 9.5 (type A) and 25.3 pS (type B), under 500 mM KCl symmetric salt conditions, whereas various conductances, ranging from 5-20 pS, were observed in the tetramethylether analogue reported previously. Results indicate that the replacement of four methylether groups with hydroxyls at the 7, 7', 12, and 12' positions in the bischolic acid framework influences the stability of aggregated (supramolecular) ion channel structures. Ion permeability ratios (potassium/chloride and potassium/sodium) for each type of channel were also determined. PMID- 15351827 TI - Synthesis of chlorinated and non-chlorinated biphenyl-2,3- and 3,4-catechols and their [2H3]-isotopomers. AB - A synthetic scheme is described for chlorinated biphenyl-2,3- and 3,4-catechols to be used as standards for structural assignment of metabolites and protein adducts of 2,2',5,5'-tetrachlorobiphenyl in which both rings retain chlorine substituents. The scheme has general applicability to the synthesis of chlorinated biphenyl catechols. Dimethyl catechol ethers are coupled to dichloroaniline via the Cadogan reaction to give a library of isomers, followed by demethylation of the ethers with BBr3 to yield the target catechols. Separation of pure isomers is accomplished by TLC or HPLC prior to or following demethylation, depending on the isomer mixture. [2H3]-Isotopomers are generated using 2,5-dichloroaniline-d3 as the starting arylamine in the coupling reaction. The dichloroaniline-d3 hydrochloride is obtained as the sole product from nitration of p-dichlorobenzene-d4 followed by Pd/C-catalyzed hydrogenation under strongly acidic conditions. This hydrogenation procedure provides a simple and convenient approach to selective reduction of aryl nitro groups in the presence of halogen ring substituents. PMID- 15351828 TI - Asymmetric total synthesis of sperabillins B and D via lithium amide conjugate addition. AB - Diastereoselective conjugate addition of homochiral lithium (R)-N-allyl-N-alpha methylbenzylamide to methyl (2E,5E)-hepatadienoate, followed by protecting group manipulation and subsequent iodocyclocarbamation allows a concise route to the core fragment, methyl (3R,5R,6R)-3,6-diamino-5-hydroxyheptanoate, of sperabillins B and D. Differentiation between the C-3 and C-6 primary amino groups of this core amino acid was readily achieved by treatment with acetone, giving the 5,6 isopropylidene and C-3-imine protected diamine, with subsequent regioselective acylation of the C-6-nitrogen facilitating the total synthesis of sperabillin D in 10.8% overall yield, and the first asymmetric synthesis of sperabillin B in 5.8% overall yield. PMID- 15351829 TI - Enzymatic cyclization reactions of geraniol, farnesol and geranylgeraniol, and those of truncated squalene analogs having C20 and C25 by recombinant squalene cyclase. AB - The substrate specificity of squalene-hopene cyclase was investigated using the C10-C25 analogs including naturally occurring substances, e.g. geraniol (C10), farnesol (C15) and geranylgeraniol (C20). No cyclization occurred for geraniol, but a significantly high conversion ratio (64%) was observed for farnesol, yielding the cyclic sesquiterpenes consisting of 6/6-fused bicyclic ring systems. Among them, an attractive compound having C30 was produced, in the structure of which acyclic the farnesol unit is linked to the bicyclic skeleton through ether linkage. Conversion of geranylgeraniol was low (ca. 12%). The squalene analogs having C20 and C25 also were cyclized in yields of ca. 33-36%, but the analogs having the methyl group at C7 and/or at C11 underwent no cyclization; the large steric bulk size of C7-Me and/or C11-Me, which is arranged in [small alpha] disposition for all the pre-chair conformation, would have interacted repulsively with the cyclase recognition site near to the C7 and/or C11, resulting in no construction of the all-chair conformation inside the reaction cavity. A relatively low yield of geranylgeraniol indicated that a less bulky hydrogen atom must be located at C14 for the efficient polycyclization reaction. The squalene cyclase shows remarkably broad substrate specificity to accept the truncated analogs having carbon-chain lengths of C(15)-C25 in addition to C30. PMID- 15351830 TI - Ring-closing metathesis for the synthesis of side chain knotted pentapeptides inspired by vancomycin. AB - A versatile method for the synthesis of bicyclic side chain knotted peptides inspired by vancomycin is described. The synthetic approach is based on the incorporation of a central amino acid derivative having two allylic groups introduced by a Stille coupling-into pentapeptide 8 containing two allylated serine residues. Treatment of this bis-ring-closing metathesis precursor with 2nd generation Grubbs catalyst results in the formation of a bicyclic pentapeptide with the correct side chain to side chain connectivity pattern as observed in vancomycin: i- 2 --> i, i --> i + 2. Modelling studies using MacroModel hint at a cavity-like structure of the bicyclic pentapeptide which may bind suitable ligands. PMID- 15351831 TI - The synthesis of unsymmetrically N-substituted chiral 1,4,7-triazacyclononanes. AB - A number of chiral unsymmetrically N-substituted 1,4,7-triazacyclononane ligands have been prepared by modular methods. The key step in the synthesis centres on the macrocyclisation of three tertiary amide precursors under standard Richman Atkins conditions which allows for subsequent N-functionalisation. PMID- 15351832 TI - Nucleophilic alkylations of 3-nitropyridines. AB - 3-Nitropyridine and 4-substituted-3-nitropyridines were reacted with chloroform, methyl chloroacetate and ethyl 2-chloropropionate under vicarious nucleophilic substitution (VNS) conditions. Substitution was obtained in the ortho or para position to the nitro group with acceptable to good yields and regioselectivity. With potassium 5-nitropyridine-2-sulfonate the substitution took place in the 4 position. Further substitution of the sulfonate group proved to be possible. PMID- 15351833 TI - An empirical study of the effect of the variables in a flash vacuum pyrolysis (FVP) experiment. AB - The effect of the variation of the experimental parameters on the conversion of precursor to products in a typical flash vacuum pyrolysis (FVP) experiment was investigated empirically. Temperature-conversion plots can be used to optimise FVP conditions and their mechanistic significance is exemplified. At a given temperature, the conversion can be increased by an increase in the background pressure, or by packing a section of the furnace tube with inert material (particularly when placed at the trap end of the furnace tube) or by employing a catalyst. Despite the prevailing view that only intramolecular reactions take place by FVP, it has been shown by a 'dual-FVP' cross-over experiment that the dimerisation of benzyl radicals occurs in the gas-phase, before the cold trap, under standard conditions. However, reduction in through-put rate, increase in furnace temperature and reduction in background pressure all reduce the amount of gas-phase coupling. PMID- 15351834 TI - Enantioselective synthesis of non-natural amino acids using phenylalanine dehydrogenases modified by site-directed mutagenesis. AB - The substrate scope of three mutants of phenylalanine dehydrogenase as biocatalysts for the transformation of a series of 2-oxo acids, structurally related to phenylpyruvic acid, to the analogous alpha-amino acids, non-natural analogues of phenylalanine, has been investigated. The mutant enzymes are more tolerant than the wild type enzyme of the non-natural substrates, especially those with substituents at the 4-position on the phenyl ring. Excellent enantiocontrol resulted in all cases. PMID- 15351835 TI - Effect of molecular symmetry on melting temperature and solubility. AB - Molecular symmetry has a pronounced effect on the melting properties and solubility of organic compounds. As a general rule, symmetrical molecules in crystalline form have higher melting temperatures and exhibit lower solubilities compared with molecules of similar structure but with lower symmetry. Symmetry in a molecule imparts a positive amount of residual entropy in the solid phase (i.e., more possible arrangements leading to the same structure). This means that the entropy of a crystal of symmetric molecules is greater than the entropy of crystal of a similar, but non-symmetric molecule. An analysis is presented relating the enthalpy, entropy and temperature of melting for an idealised system of structural isomers of different molecular symmetries. The analysis presented helps explain why often, yet not always, the crystal of a more symmetric molecule, which has greater entropy to start (closer to that of the liquid), also exhibits a greater gain in entropy upon melting, compared with the crystal of a less symmetrical molecule. The residual entropy due to molecular symmetry has the direct effect of reducing the entropy gain upon melting (a negative effect). However, molecular symmetry also exerts indirect effects on both the entropy and enthalpy of melting. These indirect effects, imposed by the condition of equilibrium melting, are positive, such that it is the balance between the direct and indirect effects what determines the value observed for the entropy of melting of the symmetric molecules. When the indirect effect of molecular symmetry is greater than its direct effect, the observed entropy gain upon melting of the more symmetrical molecule is greater than that of a less symmetrical one. PMID- 15351836 TI - Cell specific control of coagulation and fibrinolysis within the lung. PMID- 15351837 TI - Antiangiogenic properties of low molecular weight heparin - does tissue factor provide the answer? PMID- 15351838 TI - Mouse models in haemostasis and thrombosis. PMID- 15351839 TI - Role of tissue factor in haemostasis, thrombosis, angiogenesis and inflammation: lessons from low tissue factor mice. AB - Tissue factor (TF) contributes to a variety of biological processes by generating coagulation proteases and by the activation of protease activated receptors (PARs). Studies, using low TF mice, have provided us with novel insights into the role of TF in tissue-specific haemostasis. Low TF mice exhibit defects in the uterus, placenta, heart and lung, which are all tissues that normally express high levels of TF. We propose that these observed defects are primarily due to a reduction in the level of TF to below a critical threshold that is required to maintain adequate haemostasis. Nevertheless, a reduction in TF:FVIIa-dependent signaling may also reduce cell survival and/or compromise the integrity of the vasculature in these organs. Low TF mice are also a useful tool to study the role of TF and the coagulation cascade in other processes, such as thrombosis, inflammation and angiogenesis. PMID- 15351840 TI - Mice deficient in tissue factor demonstrate attenuated intimal hyperplasia in response to vascular injury and decreased smooth muscle cell migration. AB - Tissue factor (TF) is the primary initiator of the coagulation cascade and is thought to play a key role in the generation of arterial thrombosis. Recent studies have suggested that TF mediates inflammatory processes in the arterial wall and may be an important regulator of intimal hyperplasia. We have employed genetically engineered mice (mTF(-/-) /hTF(+)) with markedly diminished TF activity ( approximately 1% normal levels) o examine the role of TF in mediating the response to arterial injury. mTF(-/-)/hTF(+) displayed a marked reduction in intimal hyperplasia (46% decrease in intimal area, 60% decrease in intimal/medial ratio) in response to femoral artery injury when compared to wild type controls. The decreased intimal hyperplasia seen in low TF mice was noted in a model of vascular injury not associated with significant thrombosis, suggesting that it may be mediated by non-procoagulant properties of TF. Smooth muscle cells from mTF(-/-)/hTF(+) mice grew normally in response to serum, but exhibited a marked defect in cell migration in a modified Boyden chamber assay. In contrast, there was no difference in platelet derived growth factor- induced migration, suggesting that the effect of TF on smooth muscle cell migration is agonist dependent. These data suggest that TF may mediate intimal hyperplasia by regulating smooth muscle cell migration. PMID- 15351841 TI - Role of P-selectin and PSGL-1 in coagulation and thrombosis. AB - P-selectin and PSGL-1 are cell adhesion molecules, regulating the initial interactions between leukocytes and the blood vessel wall as well as between activated platelets and leukocytes. P-selectin is expressed on activated endothelial cells and platelets, while its major ligand PSGL-1 is expressed on leukocytes. Multiple studies have shown that these adhesion molecules are required for the normal recruitment of leukocytes during an inflammatory reaction. More recently, these adhesion molecules have been implicated in recruitment of leukocytes and leukocyte microparticles to thrombi. The P-selectin and PSGL-1-dependent delivery of circulating microparticles to thrombi appears to be important for normal tissue factor accumulation and fibrin generation in thrombi. PMID- 15351842 TI - Mouse models of thrombosis: thrombomodulin. AB - This review describes animal models of TM-deficiency that cause thrombosis in mice. Thrombomodulin (TM) is a key component of the protein C anticoagulant pathway by facilitating the activation of protein C by thrombin. In addition, TM integrates fibrinolytic and anti-inflammatory responses in a manner that is in part independent of protein C and thrombin. A series of genetically modified mouse strains is available in which the various and distinct functions of TM have been altered by means of site-directed mutagenesis of the TM gene locus (Thbd). The focus of the current review is the pathological activation of the hemostatic mechanism in mice with altered TM function (the pathologic activation of the hemostatic mechanism). The analysis of these mouse models has revealed novel and in part organ-specific functions of TM, most notably in the vascular bed of the placenta. In these mouse models, the severity and phenotypic expression of thrombosis is highly variable and is dependent on interaction with secondary genetic or environmental modifiers. The mutant mouse strains replicate important aspects of thrombophilia and thrombosis in humans, and provide a valuable resource to validate existing, and develop novel concepts of disease mechanisms in human patients. PMID- 15351843 TI - Dysfunctional platelet membrane receptors: from humans to mice. AB - Insights into hemostasis and thrombosis have historically benefited from the astute diagnosis of human bleeding and thrombotic disorders followed by decades of careful biochemical characterization. This work has set the stage for the development of a number of mouse models of hemostasis and thrombosis generated by gene targeting strategies in the mouse genome. The utility of these models is the in depth analysis that can be performed on the precise molecular interactions that support hemostasis and thrombosis along with efficacy testing of various therapeutic strategies. Already the mouse has proven to be an excellent model of the processes that support hemostasis and thrombosis in the human vasculature. A brief summary of the salient phenotypes from knockout mice missing key platelet receptors is presented, including the glycoprotein (GP) Ib-IX-V and GP IIb/IIIa (alphaIIb/beta3) receptors; the collagen receptors, GP VI and alpha2/beta1; the protease activated receptors (PARs); and the purinergic receptors, P2Y(1) and P2Y(12). A few differences exist between mouse and human platelets and where appropriate those will be highlighted in this review. Concluding remarks focus on the importance of understanding the power and limitations of various in vitro, ex vivo and in vivo models currently being used and the impact of the mouse strain on the described platelet phenotype. PMID- 15351844 TI - Murine thrombosis models. AB - Due to exciting advances in molecular biology, the laboratory mouse has become an important and frequently used model for studying thrombosis. This article reviews several experimental approaches that have been used to study arterial, venous, and microvascular thrombosis in mice. The advantages and limitations of different models are examined. Related topics of mouse anesthesia, phlebotomy, and in vitro hemostasis testing are also reviewed. PMID- 15351845 TI - The effect of vascular smooth muscle cell-targeted expression of tissue factor pathway inhibitor in a murine model of arterial thrombosis. AB - Tissue factor pathway inhibitor (TFPI) is a Kunitz-type protease inhibitor that regulates the extrinsic pathway of coagulation by inhibiting the factor VIIa/tissue factor (TF) catalytic complex. TFPI is expressed by both endothelial and smooth muscle cells in the vasculature and circulates at low levels. The role of local vascular TFPI in thrombosis and the development of vascular disease is unknown. To establish an experimental animal model to directly modulate smooth muscle cell-derived TFPI on the development of arterial thrombosis, transgenic mice in which a cDNA encoding murine TFPI is expressed from the murine SM22alpha promoter were generated. Expression of transgenic mRNA was 4-fold higher than the level of endogenous TFPI mRNA in arteries from transgenic mice. In situ hybridization confirmed that expression of the transgene was limited to medial vascular smooth muscle cells. Vascular TFPI activity was increased to 2 to 3-fold in carotid homogenates. There was no difference in plasma TFPI levels or hemostatic measures (PT, aPTT and tail vein bleeding times) between these mice and their wildtype littermates. In a ferric chloride-induced model of carotid thrombosis, homozygotic transgenic mice demonstrated resistance to thrombotic occlusion compared to wildtype littermates. In transgenic mice 22% occluded within 30 minutes of application while 84% of wild type mice occluded within the same time frame (p<0.01). Heterozygotic transgenic mice had an intermediate thrombotic phenotype. Taken together, these data indicated that local VSMC specific TFPI overexpression attenuated ferric chloride-induced thrombosis without systemic or hemostatic effects. Furthermore, this transgenic mouse model should prove useful for studying the role of TFPI in the development and progression of vascular disease. PMID- 15351846 TI - Targeted deletion of murine coagulation factor XII gene-a model for contact phase activation in vivo. AB - To analyze the biological role of factor XII (FXII, Hageman Factor) in vivo, we generated mice deficient for FXII using a gene targeting approach on two distinct genetic backgrounds, i.e. mixed C57Bl/6J X 129X1/SvJ and inbred 129X1/SvJ. Homozygous FXII knockout (FXII(-)/(-)) mice showed no FXII plasma activity and had a markedly prolonged activated partial thromboplastin time (aPTT). In contrast, coagulation factors XI, VIII, IX, X,VII, V, II and fibrinogen did not differ between FXII(-/-) mice and their wild-type littermates. Heterozygous matings segregated according to the Mendelian inheritance indicating that FXII deficiency does not increase fetal loss. Furthermore, matings of FXII(-/-) males and FXII(-/-) females resulted in normal litter sizes demonstrating that total FXII deficiency in FXII(-/-) females does not affect pregnancy outcome. Also, gross and histological anatomy of FXII(-/-) mice was indistinguishable from that of their wild-type littermates on both genetic backgrounds. Thus it appears that deficiency of murine FXII does not cause thrombophilia or impaired fibrinolysis in vivo. These results indicate that FXII deficiency does not affect hemostasis in vivo and we anticipate that the FXII(-/-) mice will be helpful to elucidate the biological role(s) of FXII in health and disease. PMID- 15351847 TI - The catalytic subunit of pseutarin C, a group C prothrombin activator from the venom of Pseudonaja textilis, is structurally similar to mammalian blood coagulation factor Xa. AB - Pseutarin C, a group C prothrombin activator from Pseudonaja textilis venom, is a large protein complex consisting of catalytic and nonenzymatic subunits, which are functionally similar to the mammalian FXa-FVa complex. Here, we present the complete cDNA sequence of the catalytic subunit of pseutarin C. The cDNA of the catalytic subunit encodes a protein of 449 amino acids, which includes a 22 residue signal peptide, 18-residue propeptide and a mature protein of 409 amino acids. The deduced amino acid sequence shows 74-83% identity to group D prothrombin activators from snake venom and approximately 42% identity to mammalian FX and has identical domain structure. The precursor of the catalytic subunit of pseutarin C has several unique features. The activation peptide of the catalytic subunit of pseutarin C is significantly smaller (27 as compared to 52 residues in mammalian FX) and does not contain any glycosylation sites. Unlike coagulation FXa, Ser52 and Asn45 of the light and heavy chains are O- and N glycosylated in pseutarin C catalytic sub-unit. There is a 12-residue insertion in pseutarin C catalytic sub-unit close to the region that is implicated in binding to FVa. This is the first sequence of the catalytic subunit of a group C prothrombin activator. PMID- 15351848 TI - Immunodominant T-cell epitopes in the factor VIII C2 domain are located within an inhibitory antibody binding site. AB - Formation of inhibitor antibodies to factor VIII (FVIII) is a major complication of FVIII replacement therapy for hemophilia A patients, and it occurs through a T cell dependent process. The C2 domain of FVIII contains epitopes that are recognized by antibody inhibitors. We have examined regions of the C2 domain that form epitopes for T cells in mice congenitally deficient in FVIII. We obtained CD4(+)T cells from mice immunized by intravenous infusion of therapeutic doses of recombinant human FVIII (rFVIII), or by subcutaneous injections of rFVIII or recombinant human C2 domain in adjuvant. In all cases, the T cells recognized most strongly and consistently two overlapping peptides that spanned residues 2191 to 2220 of the C2 domain. Analysis of the crystal structure of human factor VIII C2 bound to a human monoclonal antibody, BO2C11, showed these residues also constitute part of a human alloimmune B-cell epitope (Spiegel et al., Blood 2001; 98: 13-19). This region includes one of the "hydrophobic spike" protrusions, consisting of M2199 and F2200, as well as the basic residues R2215 and R2220. These residues contribute to membrane binding and to association with von Willebrand factor (vWF). These findings suggest that a major T-cell epitope in the C2 domain recognized by hemophilic mice is located within the same region that binds to inhibitors, vWF, and activated membranes. PMID- 15351849 TI - Compartment- and cell-specific expression of coagulation and fibrinolysis factors in the murine lung undergoing inhalational versus intravenous endotoxin application. AB - Intraalveolar and intravascular fibrin formation are typical hallmarks of acute inflammatory lung diseases, and may foster subsequent fibroproliferative events. We investigated the regulation and cellular sources of key coagulation and fibrinolysis factors in lungs undergoing compartmentalized challenge with endotoxin (LPS). BALB/c mice received 15 ng LPS either by intravenous injection or by inhalation. Quantitative gene expression analysis (real-time RT-PCR) was performed for tissue factor (TF), TF pathway inhibitor (TFPI), tissue-type plasminogen activator (t-PA), urokinase-type-PA (u-PA), PA inhibitor-1 (PAI-1), and PAI-2 in peripheral white blood cells (PBC) as well as in alveolar macrophages (AM), type-II pneumocytes (ATII), endothelial cells (EC) and smooth muscle cells (SMC), all obtained by laser microdissection. Neither route of LPS administration caused substantial protein leakage or leukocyte recruitment into the alveolar space. Compartmentalized upregulation of procoagulant and downregulation of fibrinolytic activities was, however, observed in response to both modes of LPS challenge. Intraalveolar endotoxin, in particular, caused strong upregulation of TF ( approximately 20-fold increase in gene expression) and PAI-2 (225-fold increase) in microdissected AM, upregulation of PAI-1 in microdissected ATII (300-fold increase) and EC (180-fold increase), upregulation of t-PA in EC (40-fold), and downregulation of u-PA in vascular smooth muscle cells. TFPI was largely unchanged in all cell types, and PBC showed no major gene regulatory response to inhaled endotoxin. We conclude that the lung possesses a cell-specific alveolar coagulation and fibrinolysis system, being independent of the vascular coagulation cascade and responding readily with enhanced procoagulant and anti-fibrinolytic activities to LPS challenge. PMID- 15351850 TI - Influence of polymorphisms in the factor VII gene promoter on activated factor VII levels and on the risk of myocardial infarction in advanced coronary atherosclerosis. AB - In this study, we investigate the influence of three factor VII (FVII) gene polymorphisms on activated FVII levels (FVIIa), and also on the risk of myocardial infarction (MI) in patients with advanced coronary atherosclerotic disease (CAD). The -323A2 allele in the promoter is known to be associated with low FVII levels, and has been suggested to protect against MI in some studies. The -402GA promoter polymorphism, that in vitro has been associated with having opposite effect, is less well studied clinically. For this study, plasma FVIIa levels and three FVII gene polymorphisms were assessed in 934 subjects of both sexes, all with an angiographic documentation of coronary vessels. Our results show that two promoter polymorphisms, plasma cholesterol, and gender, were significant predictors of FVIIa levels. The -402A allele was associated to a significant increase of FVIIa levels in males (by 19.2%). In a selected clinical model including the patients with severe CAD, with or without a thrombotic complication (MI), male carriers of the -402A had an increased risk of MI (OR=1.79; 95% CI 1.15-2.80). The -323A2 allele was associated to a significant decrease in FVIIa (by 36.02% in males, and 39.7% in females). Male carriers of the -323A2 were protected from MI (OR=0.6; 95% CI 0.39-0.94), but only after correction for the confounding effect of combined heterozygosity for the promoter polymorphisms. We can conclude that FVII gene polymorphisms with an opposite effect on FVIIa levels may modulate the risk of MI in males with advanced CAD. This study highlights a "within-gene" interaction, and the need to explore polymorphisms in candidate gene(s) in detail. PMID- 15351851 TI - Antithrombin reduces ischemia/reperfusion-induced liver injury in rats by activation of cyclooxygenase-1. AB - This study was conducted to determine which isoform of cyclooxygenase (COX) is more significantly involved in the anti-thrombin (AT)-induced increase in prostaglandin production in the liver of rats, subjected to hepatic ischemia/reperfusion (I/R). Hepatic tissue levels of 6-keto-PGF(1alpha), a stable metabolite of prostacyclin (PGI(2)), and PGE(2) were transiently increased 1 hour after reperfusion. Thereafter, hepatic PGE2 levels were gradually increased until 6 hours after reperfusion, while hepatic 6-keto-PGF(1alpha) levels were decreased to the pre-ischemia levels at 6 hours after reperfusion. AT significantly enhanced increases in hepatic tissue levels of 6-keto-PGF(1alpha) and PGE(2) seen 1 hour after reperfusion, while it inhibited increases in hepatic PGE(2) levels seen 6 h after reperfusion. Neither dansyl-Glu-Gly-Arg-chloromethyl ketone treated factor Xa (DEGR-Xa), a selective inhibitor of thrombin generation, nor Trp(49)-modified AT which lacks affinity for heparin, showed any effects on these changes. Pretreatment with indomethacin (IM), a non-selective inhibitor of COX, inhibited AT-induced increases in hepatic tissue levels of 6-keto-PGF(1alpha) and PGE(2) seen 1 hour after reperfusion, whereas pretreatment with NS-398, a selective inhibitor of COX-2, did not. The increase in hepatic tissue blood flow and inhibition of hepatic inflammatory responses seen in animals given AT were reversed by pretreatment with IM, but were not affected by pretreatment with NS 398. Administration of ilo-prost, a stable analog of PGI(2), and PGE(2) produced effects similar to those induced by AT. Increases in hepatic tissue levels of PGE(2) 6 hours after reperfusion were inhibited by pretreatment with NS-398. Although AT did not affect COX-1 mRNA levels 1 hour after reperfusion, it inhibited the I/R-induced increases in hepatic tissue levels of both PGE(2) and COX-2 mRNA 6 hours after reperfusion. These observations strongly suggested that AT might reduce the I/R-induced liver injury by increasing the production of PGI2 and PGE2 through activation of COX-1. Furthermore, since TNF-alpha is capable of inducing COX-2, inhibition of TNF-alpha production by AT might inhibit COX-2 mediated PGE(2) production. These effects induced by AT might contribute to its anti-inflammatory activity. PMID- 15351852 TI - A comparative study of amyloid-beta (1-42) as a cofactor for plasminogen activation by vampire bat plasminogen activator and recombinant human tissue-type plasminogen activator. AB - The activity of both human tissue-type plasminogen activator (t-PA) and the PA from the saliva of the vampire bat, Desmodus rotundus, (DSPA) is critically dependent on the presence of a cofactor. The most efficient cofactor for both PAs is fibrin, but fibrinogen and amyloid beta peptides also have cofactor activities for human t-PA. Compared to t-PA, DSPA has a more stringent requirement for fibrin as a cofactor. The present study was undertaken to compare cofactor activities of amyloid beta 1-42 (Abeta1-42) for plasminogen activation by DSPA alpha1 or by t-PA. The two PAs were incubated with different concentrations of glu-plasminogen, a chromogenic substrate for plasmin and 100 micro g mL (-1) of Abeta1-42, fibrinogen or fibrin as cofactor. Using the kinetic parameters directly determined from the chromogenic substrate conversion curves, we derived the relative efficacies of DSPA or t-PA in the presence of cofactor at the physiological plasminogen concentration of 2 micro M. In the presence of fibrin, the activity of DSPA was comparable to that of t-PA and 23,270-fold higher than its activity without cofactor, whereas fibrin induced only a 248-fold increase in t-PA activity. The activity of DSPA with Abeta1-42 or fibrinogen as cofactor was 485-fold lower than its activity in the presence of fibrin, while for t-PA this difference was only 26-fold. The much lower activity of DSPA as compared to t-PA with Abeta1-42 or fibrinogen might lead to fewer side effects when used for the thrombolytic therapy of stroke. PMID- 15351853 TI - Activation of fibrinolysis in the pericardial cavity after cardiopulmonary bypass. AB - Aprotinin is frequently administered systemically in patients undergoing cardiopulmonary bypass to preserve platelet function and ameliorate excessive activation of fibrinolysis. More recently, aprotinin topically applied in the pericardial cavity was also found to improve postoperative blood loss. However, platelet activation was not reduced locally during surgery. Hence, we investigated in the present prospective, in a randomized double-blind trial, the intra- and early postoperative state of systemic and local fibrinolytic activity, and whether topically administered aprotinin acts as an antifibrinolytic and therefore improves local hemostasis. Patients undergoing elective coronary artery bypass grafting were divided in two groups containing 22 patients each. Both, group I and II patients received high-dose aprotinin (6.0 x 10(6) kallikrein inhibitor units (KIU)) systemically. Before resuming extracorporeal circulation (ECC), either 1.0 x 10(6) KIU aprotinin (group I) or vehicle solution (group II) was applied into the pericardial cavity. Plasminogen, 2 alpha(2)-antiplasmin, plasmin-alpha(2)-antiplasmin complex, plasminogen-activator-inhibitor type-1 and D-dimers were measured in the pericardial cavity and systemic circulation immediately before resuming extracorporeal circulation (ECC), and at 1 h and 4 h postoperatively. The local fibrinolytic activity was found to significantly exceed that measured in the systemic circulation over time, whether ot not they received aprotinin into the thoracic wound surface. Furthermore, evidence was provided that topically applied aprotinin reduces not only the local fibrinolytic activity but also the postoperative blood loss significantly by 33% which demonstrates the clinical relevance. The local activation of fibrinolysis seems to play an important role in blood loss after cardiopulmonary bypass. Therefore in fibrinolysis and blood coagulation the surgeon should not only consider what happens in the systemic circulation but also on a local level. PMID- 15351854 TI - Recombinant urokinase for restoration of patency in occluded central venous access devices. A double-blind, placebo-controlled trial. AB - The interval occlusion of central venous access devices (CVADs) remains a significant clinical problem, often requiring re-intervention for catheter exchange or replacement. The purpose of this Phase 3, multi-center, double blinded study was to test the hypothesis that instillation of recombinant urokinase (r-UK) 5000 IU/ml is superior to placebo in restoring total catheter patency to an unselected cohort of occluded CVADs. After obtaining informed consent, adult and pediatric patients with occluded, non-hemodialysis CVADs of any duration or type were randomized (2 : 1) to receive either r-UK 5000 IU/ml or placebo instilled into all occluded lumens of their catheter. Catheter function was assessed at 5, 15 and 30 min after the first instillation. If the catheter remained occluded after 30 min, a second dose was instilled with repeat assessments at 5, 15 and 30 min. The primary efficacy variable was the restoration of catheter function to all treated lumens (i.e., total catheter patency) after one or two instillations. Catheters that were not successfully recanalized after two instillations were allowed to receive up to two instillations of open-label r-UK administered in the same manner. The primary safety variable was the occurrence of hemorrhagic and non-hemorrhagic events within 72 hr after instillation. A total of 180 patients were enrolled at 43 sites in the United States and Canada. Most patients were adults, although 20% were Thr mutation in the transmembrane region of GP IX does not induce intracellular GP Ib/IX complex degradation, but prevents its insertion in the cytoplasmic membrane of platelets and CHO cells. PMID- 15351859 TI - ADP induced platelet degranulation in healthy individuals is reduced by clopidogrel after pretreatment with atorvastatin. AB - Statins inhibit platelet reactivity and reduce blood thrombogenicity. The effectiveness of clopidogrel in inhibiting platelet reactivity was suggested to be reduced in the presence of atorvastatin due to shared enzymes in metabolism. Healthy individuals, 17 pretreated with atorvastatin (20 mg/d for 3 days) and 17 without pretreatment, as well as 15 patients with stable coronary artery disease (CAD) and concurrent atorvastatin therapy were started on clopidogrel (loading dose 300 mg, then 75 mg/d). P-selectin on platelet surface after stimulation with ADP or Thrombin Receptor Activating Peptide (TRAP) was flow cytometrically measured before and during clopidogrel administration. Whole blood platelet agglutination was tested by a platelet function assay.TRAP and - in trend - ADP induced p-selectin exposure was reduced by the atorvastatin pretreatment before clopidogrel was added. Combining clopidogrel with atorvastatin in the healthy individuals led to a further reduction in ADP-induced platelet p-selectin exposure. Clopidogrel also reduced platelet reactivity in CAD patients with concurrent atorvastatin medication. We conclude that pretreatment with atorvastatin reduces platelet reactivity before administration of clopidgrel. No drug interaction was seen, however, platelet inhibitory effects were observed during the treatment with clopidogrel and atorvastatin. PMID- 15351860 TI - Platelet hyperreactivity in hemodialysis patients with frequently occluded vascular access. AB - It is known that thrombosis is a leading cause of vascular access failure and that the formation of thrombus requires platelets. The activation of platelets induces the increase in intracellular Ca 2(+) levels ([Ca(2+)](i)) leading to aggregation and thrombosis. We compared the platelet [Ca(2+)](i) before and after stimulation between the patients with and without easily occluded vascular access. Our study included two groups of hemodialysis patients. Group 1 consisted of 21 patients who had received chronic hemodialysis therapy for more than 6 months. They had had more than three events (including three) of vascular access failures during the past year. Group 2 consisted of 21 hemodialysis patients with age, sex, and diabetes mellitus matched who had never suffered from any event of vascular access failure. We measured the basal and stimulated platelet [Ca(2+)](i) after stimulation with 1 U/ml thrombin, 1 micro M arachidonic acid, 1 micro M platelet activation factor (PAF), and 10 micro M adenosine diphosphate (ADP), respectively. Our results showed that in Ca 2(+)-containing media, there was no significant differences in the basal [Ca(2+)](i), but the maximal increases of [Ca(2+)](i) of platelets were higher (p <0.05) in group 1 than in group 2 after stimulating with PAF and ADP, but not with thrombin and arachidonic acid. We concluded that the causes for the susceptibility of some hemodialysis patients to vascular access occlusion were multifactorial. In addition to previously reported plasma factors, there was a sub-group of patients who showed greater elevations of agonists stimulated platelet intracellular calcium levels. PMID- 15351861 TI - Inhibition of endothelial cell tube formation by the low molecular weight heparin, tinzaparin, is mediated by tissue factor pathway inhibitor. AB - Heparin and low molecular weight heparins (LMWHs) have both antithrombotic and anti-angiogenic activities. The anti-angiogenic activity of LMWH may be associated with the release of endothelial tissue factor pathway inhibitor (TFPI), an important endogenous inhibitor of tissue factor/Factor VIIa (TF/fVIIa). To evaluate the effects of LMWH, tinzaparin, and TFPI in a model of angiogenesis-mediated processes, we compared the effects of tinzaparin, and recombinant TFPI in inhibiting either basic fibroblast growth factor-2 (FGF2)- or TF/fVIIa-induced endothelial cell tube formation in human umbilical vein endothelial cells (HUVEC). Our results show that tinzaparin and recombinant TFPI both blocked endothelial tube formation induced by either FGF2 or TF/fVIIa, in a concentration-dependent manner. Endothelial tube formation was only marginally inhibited by a potent and specific anti-Factor Xa, recombinant tick anticoagulant protein (rTAP). A monoclonal anti-TFPI anti-body reversed the inhibitory effects of either tinzaparin or recombinant-TFPI on HUVEC tube formation. Tinzaparin fractions in the range of 8,000 to 12,600 Da were most effective in stimulating the release of TFPI from HUVEC. These results suggest that the inhibitory effect of the LMWH tinzaparin on endothelial tube formation is associated with stimulation of the release of TFPI, but not to anti-Factor Xa activity. PMID- 15351862 TI - Modulation of VEGF-induced endothelial cell cycle protein expression through cyclic AMP hydrolysis by PDE2 and PDE4. AB - Endothelial cell proliferation in response to VEGF plays an important role in physiological and pathological angiogenesis. The role of PDE2 and PDE4 in VEGF induced proliferation in HUVEC was investigated: 1) VEGF increased cAMP hydrolytic activity by up-regulating the expression of PDE2 and PDE4 isozymes; 2) VEGF increased progression in cell cycle with an increase in p42/p44 MAP kinase, cyclin A and cyclin D1 expressions and with a decrease in p21 waf1/cip1 and p27 kip1 expressions; 3) EHNA (20 micro M), a selective PDE2 inhibitor, RP73401 (10 micro M), a selective PDE4 inhibitor blocked the VEGF-induced increase in p42/p44 MAP kinase expression; 4) RP73401, but not EHNA, blocked the VEGF-induced increase in cyclin A and decrease in p27 kip1 expressions; 5) EHNA, contrary to RP73401, enhanced the VEGF-induced increase of cyclin A and decrease of p27 kip1. 6) EHNA and RP73401 together blocked the VEGF-induced increase in cyclin D1 and decrease in p21 waf1/cip1 expressions; 7) Inhibition of VEGF-upregulated PDE2 and PDE4 reversed the VEGF-induced alterations in cell cycle protein expression, bringing back endothelial cells to a non-proliferating status. Consequently, PDE2 and PDE4 inhibitions were able to inhibit VEGF-induced endothelial cell proliferation by restoring cell cycle key protein expression, and might thus be useful in excessive angiogenesis. Furthermore, the differences between PDE2 and PDE4 effects may suggest compartmentalized effects. PMID- 15351863 TI - Extrinsic coagulation pathway activation and metalloproteinase-2/TIMPs system are related to oxidative stress and atherosclerosis in hemodialysis patients. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), together with extrinsic coagulation pathway activation and increased oxidative stress (SOX) have all been implicated as important factors in atherosclerosis and vascular remodelling. The aim of the present study was to investigate whether the MMPs/TIMPs system is associated with activation of the extrinsic coagulation pathway in conditions of increased SOX in hemodialysis (HD) patients. In HD patients, with and without cardiovascular disease (CVD), and in controls, we compared pre-dialysis levels of MMP-2, MMP-9,TIMP-1,TIMP-2; the markers of extrinsic coagulation pathway-tissue factor (TF) and its inhibitor (TFPI), prothrombin fragment F1+2 (F1+2); a marker of SOX-Cu/Zn superoxide dismutase (Cu/Zn SOD) and a surrogate of atherosclerotic disease-intima media thickness (IMT). Hemodialysis patients, particularly those with CVD, showed a significant increase in values of MMP-2/TIMPs, markers of the extrinsic coagulation pathway, Cu/Zn SOD and IMT as compared to controls. The markers of coagulation activation positively correlated with the MMP-2/TIMPs system, whereas they did not correlate with MMP-9. In addition, both MMP-2/TIMPs as well as extrinsic coagulation parameters were related to the prevalence of increased SOX, IMT and CVD. Multiple stepwise regression analysis showed that low TIMP-2 followed by increased Cu/Zn SOD and MMP-2 levels independently and significantly predicted elevated IMT on maintenance HD patients. In conclusion, our data suggest that the MMP-2/TIMPs system and an activated extrinsic coagulation pathway could cooperate in conditions of elevated SOX and could influence arterial remodeling resulting in the presence of CVD in haemodialysis patients. PMID- 15351865 TI - Does haemodilution produce a hypercoagulable state? PMID- 15351864 TI - Prevention of central venous line-related thrombosis by continuous infusion of low-dose unfractionated heparin, in patients with haemato-oncological disease. A randomized controlled trial. AB - We have conducted a prospective randomized controlled trial to evaluate the role of low-dose unfractionated heparin prophylaxis in preventing central venous line related thrombosis in patients with haemato-oncological disease. Patients were randomly assigned to receive either prophylactic intravenous unfractionated heparin (continuous infusion of 100 IU/kg/daily) or 50 ml/daily of normal saline solution as a continuous infusion. CVLs were externalized, non tunneled, double lumen catheters. All CVLs were placed percutaneously by the same physician in the subclavian vein. Upper limb veins were systematically examined by ultrasonography just before, or <24 hours after, catheter removal, and in case of clinical signs of thrombosis. One hundred and twenty-eight CVLs were inserted. Catheter-related thrombosis occurred in 1.5% of the catheters inserted in patients of the heparin group, and in 12.6% in the control group (p = 0.03). No other risk factors were found for the development of catheter-related thrombosis. Two and three patients experienced severe bleeding in the heparin group, and in the control group, respectively (p = 0.18). There were no other side-effects clearly ascribable to the use of unfractionated heparin. This is the first prospective, randomized study, which shows that low-dose of unfractionated heparin is safe and effective to prevent catheter-related thrombosis in patients with haemato-oncological disease. PMID- 15351866 TI - [Patterns on the skin. New aspects of their embryologic and genetic causes]. AB - The concept of cutaneous mosaicism has now been proven by numerous studies. In this way, the idea that all nevi represent mosaics has so far been confirmed. We can distinguish five different patterns of mosaicism in human skin in the form of lines of Blaschko, checkerboard pattern, phylloid pattern, patchy pattern without midline separation, and lateralization. Two different etiological categories are epigenetic and genomic mosaicism. According to present knowledge all epigenetic mosaics are caused by the activity of retrotransposons. In addition to the well known examples of functional X-chrosomosome mosaicism, the concept of autosomal functional mosaicism, as observed in mice and dogs, has recently been established. Most likely, this mechanism may also explain the exceptional familial occurrence of pigmentary mosaicism in man. Phylloid hypomelanosis and speckled lentiginous nevus syndrome are recently described phenotypes that can be added to the list of lethal mutations surviving by mosaicism. The concept of type 2 segmental manifestation of autosomal dominant skin diseases has recently been confrirmed at the molecular level in a case of Hailey-Hailey disease. The concept of didymosis may explain why two different nevi can occur in spatial and temporal proximity, e.g., cutis tricolor or phacomatosis pigmentovascularis. The mechanism of paradominant inheritance may explain why some nevi that usually occur sporadically may affect, by way of exception, several members of a family. Examples are sebaceous nevus and Becker nevus. In autosomal recessive skin disorders, an event of loss of homozygosity may occur at an early developmental stage, giving rise to healthy skin areas arranged in a nevoid distribution. For example, when dermatologists examine a patient with xeroderma pigmentosum they should pay particular attention to such areas of revertant mosaicism. PMID- 15351867 TI - [Purely cutaneous Rosai-Dorfman disease]. AB - Sinus histiocytosis with massive lymphadenopathy or Rosai-Dorfman disease is a non-Langerhans cell histiocytosis of unknown etiolosy. The most characteristic feature is lymphadenopathy, especially that of cervical lymph nodes. In approximately 40% of patients there are extranodal manifestations of the disease. Skin is the most commonly affected organ (27%). We report a 79-year-old female patient with purely cutaneous manifestations. This form of the disease is a very rare differential diagnosis in dermatology. PMID- 15351868 TI - [Mucosal epidermolysis bullosa acquisita complicated by laryngeal stenosis]. AB - Epidermolysis bullosa acquisita (EBA) is an acquired chronic autoimmune blistering skin disease that rarely affects mucous membranes. We report a 75-year old female patient who had EBA restricted to oral, nasal and laryngeal mucous membranes with life-threatening laryngeal stenosis. PMID- 15351869 TI - [Eruptive xanthomas]. AB - A 41-year old male patient presented with yellowish papules on the elbows, upper arms, back and the buttocks, as well as yellow streaks in the palmar creases. Laboratory examination revealed Fredrickson type IIb hyperlipidemia. Histologic changes were consistent with eruptive xanthomas. Treatment was started with HMG CoA reductase inhibitors and dietary measures were taken to lower serum levels of cholesterol and triglycerides. Hyperlipidemia is the cause of eruptive xanthomas and strongly increases the risk of cardiovascular diseases. PMID- 15351870 TI - [Ulcus terebrans. Therapy options and their limits]. AB - Basal cell carcinoma (BCC) is the most common tumor in humans and is defined as a slow-growing, locally invasive, epithelial skin tumor which rarely metastasizes. The first line treatment is surgical excision with histologic examination of the tumor margins, but numerous alternative therapies are available. A 75-year old patient with the most destructive form of BCC, ulcus terebrans, involving the scalp and invading the frontal bone. We discuss the problems and therapeutic limitations for this unusual BCC variant. PMID- 15351871 TI - [Sensitivity to constituents of bone cement in a patient with joint prosthesis]. AB - Failure to tolerate prosthetic material can lead to a variety of clinical findings. A 55-year-old woman had a total replacement of the right knee. Two weeks later she developed pruritic skin lesions over the joint, as well as pain and impaired function. Allergic testing showed clinically relevant Type IV sensitization to methylmethacrylate, as well as to various formulations of the bone cement, including one with an added antibiotic. The symptom complex of pruritic skin lesions in the area of the prosthesis, pain and disability, as well as relevant Type IV sensitization justified replacing the prosthesis. PMID- 15351872 TI - Avascular necrosis of the femoral head foreshadowing familial Mediterranean fever: apropos of three cases. AB - We reported here on three patients in whom the diagnosis of familial Mediterranean fever was established after avascular necrosis of the femoral head had been detected. The pathogenesis and the management of this rare concomitance are discussed in the light of the relevant literature. PMID- 15351873 TI - Raynaud's phenomenon in undifferentiated connective tissue disease (UCTD). AB - The aim of this study was to ascertain which clinical and immunological factors are associated with Raynaud's phenomenon (RP) in patients with undifferentiated connective tissue disease (UCTD) and to investigate microvascular involvement. A total of 78 patients were evaluated. They all showed symptoms suggestive of a connective tissue disorder (CTD), but did not fulfil the criteria for any of the defined CTDs. They all had a disease duration of at least 1 year. Nailfold capillaroscopy (NC) was performed using a computerised videomicroscope. We diagnosed RP in 52.5% of our patients. Patients with RP showed a higher occurrence of oesophageal dysmotility (p=0.001) and anti-ribonucleoprotein (RNP) antibodies (p=0.004) than those without RP. The distinguishing capillaroscopic characteristics of UCTD patients with RP were widened and irregularly enlarged loops (75 and 55%, respectively), giant capillaries (35%), and less than two haemorrhages per finger (40%). The combination of features indicative of a 'slow' scleroderma pattern was present in 18 of 40 patients with UCTD and RP (p=0.0003). Only 3 of the original 78 patients (3.8%) developed a definite CTD. In none of our patients did we observe avascular areas or changes from the original capillaroscopic pattern during follow-up examination. Our study indicates that patients with UCTD would seem to have a benign form of RP, since they show the absence of cutaneous complications, the existence of a mild microvascular damage and a stable nailfold capillary pattern. Further examinations of these patients will be required in order to confirm our findings. PMID- 15351874 TI - Late bilateral diaphragmatic rupture: challenging diagnostic and surgical repair. AB - A 67-year-old man was referred to our department, after a vehicle accident, with multiple bone fractures and a left blunt diaphragmatic rupture. An emergency laparatomy was performed, and the left diaphragmatic defect directly sutured. Postoperatively, a delayed right diaphragmatic rupture occurred due to progressive inflammation and muscle devitalisation. The diagnosis was challenging because the right rupture became clinically evident later after extubation. Diaphragmatic reconstruction was performed through a right thoracotomy. A high index of suspicion should always be observed for missed or delayed bilateral diaphragmatic ruptures. PMID- 15351875 TI - A simple fluorimetric method for the estimation of DNA-DNA relatedness between closely related microorganisms by thermal denaturation temperatures. AB - Determination of whole-genome DNA-DNA similarity is today a standard technique for species delineation in microbial taxonomy. However, these studies demand hard to-perform and time-consuming experiments. Herein, we present an easy and rapid fluorimetric method to estimate DNA-DNA relatedness between microbial strains from differences of the thermal denaturation temperatures of hybrid and homologous genomic DNA. Double-stranded DNA was specifically stained with SYBR Green I, and its thermal denaturalization was followed by measuring a decrease in fluorescence. A quantitative, real-time PCR thermocycler was used to perform the experiment and obtain fluorescence determinations at increasing temperatures. The proposed method was validated by comparing species of the hyperthermophilic genera Pyrococcus and Thermococcus. The method proves to be an easy, rapid, and inexpensive alternative to estimate DNA-DNA relatedness between closely related species. PMID- 15351876 TI - Moroccan women with a history of child sexual abuse and its long-term repercussions: a population-based epidemiological study. AB - Childhood sexual abuse is defined as sexual contact imposed on a child whose development is still, from the emotional, cognitive and maturity point of view, lacking. The objective of this epidemiological study, conducted among a representative sample of the female population aged 20 and over in Casablanca, Morocco, is to determine the prevalence of the childhood sexual abuse and its long-term repercussions. The main results were that 9.2% (n=65) reported childhood sexual abuse. The mean age of this subgroup was 31.34+/-9.43 years (20 63); 66.1% were housewives; 40% were single and 10.8% were divorced. The prevalence of genital abuse with penetration was 33.8% (22 cases). The abuser was known in 56.2% and was a family member in 20.4% of the cases. Of the 728 women interviewed, 27 (43.5%) abused women and 83 (29.5%) non-abused women during childhood reported depressive symptoms with a positive association (p=0.03). There was also an association between the most severe types of abuse and vaginismus. Generally speaking, all those who were abused during their childhood suffered sexual disturbances during their adult life. PMID- 15351877 TI - Proteins interacting with the tuberous sclerosis gene products. AB - Tuberous sclerosis (TSC) is an autosomal dominant tumor suppressor gene syndrome affecting about 1 in 6000 to 10000 individuals. The genes, TSC1, encoding hamartin, and TSC2, encoding tuberin are responsible for TSC. Since their identification 1997 and 1993 respectively, a variety of different functions have been described for the TSC gene products. Hamartin and tuberin form a complex, providing a tentative explanation for the similar disease phenotype in TSC patients with mutations in either of these genes. In addition, associations of hamartin or tuberin with several different proteins have been demonstrated. In this review, we summarize the current knowledge on hamartin- and tuberin interacting proteins and discuss their role for the understanding of the functions of the TSC gene products. PMID- 15351878 TI - Peripherally inserted central venous catheter in pediatric hematological patients. PMID- 15351879 TI - Ten years' activity of the first Italian public hospice for terminally ill patients. AB - We present a review of the first 10 years of the hospice at the Geriatric Institute Pio Albergo Trivulzio of Milan, Italy's first public hospice for the admission of terminally ill patients. Over 1200 patients were admitted to the nine-bed hospice between October 1991 and December 2001, most of whom (63%) were referred by the Home Palliative Care Units operating in Milan. The hospice patients are elderly (nearly 60% are 70 or more years of age, median 72 years). Admission to the hospice was until the patient's death (74% of patients) after a brief time (4 weeks on average). From the very outset, we have striven to focus our attention on the daily application of the programmes of care inspired by the philosophy and practice of palliative medicine, i.e. the holistic approach and attention devoted to quality of life, multidimensional assessment, and the services of a multiprofessional team to provide, alongside medical and nursing assistance, psychosocial and spiritual support, bereavement support, etc. The continuing education of health workers and the systematic use of a clinical audit tool specifically designed for palliative care, are the two key elements which, in our judgement, have proved to be the most fruitful in reaching the objectives described above. These 10 years of the Pio Albergo Trivulzio Hospice have made a significant contribution towards defining a concrete Italian model which can be applied to the care of the terminally ill inpatient. PMID- 15351880 TI - A hidden cause of hypokalemic paralysis in a patient with prostate cancer. AB - Hypokalemic paralysis is a medical emergency due to the risks of cardiac arrhythmia, respiratory failure, and rhabdomyolysis. Besides supplementing patients with KCl to hasten recovery, the astute physician must search for the underlying cause to avoid missing a treatable and curable disorder. We report on an elderly Korean man who presented with marked limb paralysis, myalgias, and mild hypertension. He had prostate cancer treated with orchiectomy and hormone therapy 2 years previously. The major biochemical abnormalities were hypokalemia (K+: 1.7 mmol/l) associated with high renal K+ wasting and metabolic alkalosis (HCO3-: 42.6 mmol/l). Low plasma renin activity, low aldosterone concentration, and normal cortisol concentration pointed to a state of pseudohyperaldosteronism. While reviewing his drug history, the patient revealed he had been consuming eight packs (100 ml/pack) of a Korean herbal tonic daily to treat his prostate cancer for the past 2 months. A significant amount of glycyrrhizic acid (0.23 mg/ml), an active ingredient of licorice, was detected in the tonic. Discontinuation of the herbal tonic along with KCl supplementation achieved recovery in 2 weeks. As many complementary/alternative medicines for cancer contain licorice, this must be kept in mind as a cause of hypokalemia in cancer patients. PMID- 15351881 TI - Are endophyte-mediated effects on herbivores conditional on soil nutrients? AB - Neotyphodium endophytes are assumed to have mutualistic relationship with their grass hosts, mainly resulting from mycotoxin production increasing plant resistance to herbivores by the fungus that subsists on the plant. To study importance of often ignored environmental effects on these associations, we performed a greenhouse experiment to examine the significance of endophyte infection and nutrient availability for bird-cherry aphid ( Rhopalosiphum padi) performance on meadow fescue ( Lolium pratense). Naturally endophyte-infected (E+), uninfected (E-), or manipulatively endophyte-free (ME-) half-sib families of meadow fescue were grown on two soil nutrient levels. Endophyte infection reduced aphid performance in general. However, to our knowledge, this is the first study to demonstrate experimentally that herbivore performance decreases on E+ host plants with increasing availability of nutrients in soils. Potential improvement in herbivore performance in high nutrient soils and decreased plant performance in low nutrient soils in ME- plants, compared to E- and E+ plants, suggests that loss of endophyte infection after long coevolutionary relationship may be critical to plant fitness. PMID- 15351882 TI - Cloning, sequencing, and expression of the genomic DNA encoding the protein phosphatase cdc25 in Dictyostelium discoideum. AB - A genomic DNA ( Dd-cdc25) encoding the protein phosphatase cdc25 was isolated from the cellular slime mold Dictyostelium discoideum. The Dd-cdc25 DNA sequence, with a length of 2,958 bp, encodes a protein consisting of 986 amino acid (aa) residues. The sequence shares significant identities with cdc25 from human, mouse, Xenopus, Drosophila, and Shizosaccharomyces pombe, particularly at the C terminal region including the catalytic site for phosphatase activity. The deduced Dictyostelium cdc25 protein (Dd-cdc25) has the highest molecular mass (109.9 kDa) in several cdc25 species so far reported and contains four regions consisting of unusually long asparagine repeats (22-31) in the sequence. Unexpectedly, however, Western blot analysis using a specific antibody raised against the C terminus (aa 892-986) of Dd-cdc25 demonstrated that the protein exists as a short form (56 kDa), which has the C-terminal active site of phosphatase, during the course of Dictyostelium development. The Western blot analysis also revealed marked changes in the phosphorylated state of the Dd cdc25, coupling with cellular development. PMID- 15351883 TI - Expression of poly-3-(R)-hydroxyalkanoate (PHA) polymerase and acyl-CoA transacylase in plastids of transgenic potato leads to the synthesis of a hydrophobic polymer, presumably medium-chain-length PHAs. AB - Medium-chain-length poly-3-(R)-hydroxyalkanoates (mcl-PHAs) belong to the group of microbial polyesters. The minimum gene-set for the accumulation of mcl-PHAs from de novo fatty acid biosynthesis has been identified in prokaryotes as consisting of the Pha-C1 polymerase and the ACP-CoA-transacylase. In this paper, the synthesis of mcl-PHAs has been attempted in transgenic potato (Solanum tuberosum L.) using the same set of genes that were introduced into potato by particle bombardment. Polymer contents of transgenic lines were analysed by gas chromatography and by a new simple method employing a size-exclusion filter column. The expression of the Pha-C1 polymerase and the ACP-CoA-transacylase in the plastids of transgenic potato led to the synthesis of a hydrophobic polymer composed of mcl-hydroxy-fatty acids with carbon chain lengths ranging from C-6 to C-12 in leaves of the selected transgenic lines. We strongly suggest that the polymer observed consists of mcl-PHAs and that this report establishes for the first time a possible route for the production of mcl-PHAs from de novo fatty acid biosynthesis in plants. PMID- 15351884 TI - The Nissen fundoplication: indication, technical aspects and postoperative outcome. AB - INTRODUCTION: Gastroesophageal reflux disease (GERD) is the most common foregut disease, with a great impact on quality of life and with intestinal, respiratory and cardiac symptoms and implications of carcinogenesis of the oesophagus. Medical therapy often fails, due to the complex pathophysiology of GERD. Surgery can cure the disease, since it is able to restore the anti-reflux barrier. It improves quality of life and prevents carcinogenesis. METHODS: Review of the literature and presentation of our own experience and data in a series of more than 4,000 evaluated patients referred for suspected reflux disease, of whom 382 have been operated on. CONCLUSION: The laparoscopic Nissen fundoplication is the most commonly used operation technique. It provides good long-term results in the majority of patients. However, due to an increase of outflow resistance of the oesophagus this operation is associated with some postoperative side effects. Therefore, alternative anti-reflux procedures may be indicated in selected patients. PMID- 15351885 TI - Laparoscopic appendicectomy is associated with a lower complication rate even during the introductory phase. AB - BACKGROUND: The benefit of laparoscopic appendicectomy is under debate. To study the beneficial clinical effects of laparoscopic appendicectomy during the introductory phase we analysed, retrospectively, 493 patients from one district general hospital. PATIENTS AND METHODS: During a period of 3 years 250 patients were operated on prospectively by open appendicectomy, and 243 patients were operated on laparoscopically. Both groups were compared with regard to their demographic data, operation time, body mass index and complication rate. RESULTS: The conversion rate was 4.5%. The distribution of histological stages of inflammation was comparable in both groups. The median operating time was significantly longer for the open (40 min) than for the laparoscopic procedure (35 min, P=0.002). The body mass index in the laparoscopic group was significant higher (23.7 kg/m(2) vs 22.6 kg/m(2), P=0.009). Perioperative white blood cell count, C-reactive protein and body temperature were equal in both groups. There was no significant difference with regard to the analgesia required postoperatively between the open and the laparoscopic group. Nevertheless, the specific complication rate after open appendicectomy (18.4%) was significantly higher than that following laparoscopic appendicectomy (10.8%, P=0.03). CONCLUSION: Even during the introductory phase, laparoscopic appendicectomy is a safe and clinically beneficial operating procedure. PMID- 15351886 TI - A general model for visual motion detection. AB - We propose a general model for detection of both first-order motion and second order motion. In this model an input stimulus is divided into a number of partially overlapping spatiotemporal local regions. Spatiotemporal frequency analysis is done for every local region using Gabor filters, then the input stimulus (original spatiotemporal signal) is replaced by the outputs of Gabor filters. Local motion is detected by applying Gabor motion detectors to each local spatiotemporal pattern depicted by each local feature value. Outputs of all the detectors are integrated to give the final output for global motion of the input stimulus. The model was simulated on a computer and was confirmed to correctly detect second-order motion as well as first-order motion. PMID- 15351887 TI - Biomechanical mechanism for transitions in phase and frequency of arm and leg swing during walking. AB - As humans increase walking speed, there are concurrent transitions in the frequency ratio between arm and leg movements from 2:1 to 1:1 and in the phase relationship between the movements of the two arms from in-phase to out-of-phase. Superharmonic resonance of a pendulum with monofrequency excitation had been proposed as a potential model for this phenomenon. In this study, an alternative model of paired pendulums with multiple-frequency excitations is explored. It was predicted that the occurrence of the concurrent transitions was a function of (1) changes in the magnitude ratio of shoulder accelerations at step and stride frequencies that accompany changes in walking speed and (2) proximity of these frequencies to the natural resonance frequencies of the arms modeled as a pair of passive pendulums. Model predictions were compared with data collected from 14 healthy young subjects who were instructed to walk on a treadmill. Walking speeds were manipulated between 0.18 and 1.52 m/s in steps of 0.22 m/s. Kinematic data for the arms and shoulders were collected using a 3D motion analysis system, and simulations were conducted in which the movements of a double-pendulum system excited by the accelerations at the suspension point were analyzed to determine the extent to which the arms acted as passive pendulums. It was confirmed that the acceleration waveforms at the shoulder are composed primarily of stride and step frequency components. Between the shoulders, the stride frequency components were out-of-phase, while the step frequency components were in-phase. The amplitude ratio of the acceleration waveform components at the step and stride frequencies changed as a function of walking speed and were associated with the occurrence of the transitions. Simulation results using these summed components as excitatory inputs to the double-pendulum system were in agreement with actual transitions in 80% of the cases. The potential role of state-dependent active muscle contraction at shoulder joints on the occurrence of the transitions was discussed. Due to the tendency of arm movements to stay in the vicinity of their primary resonance frequency, these active muscle forces were hypothesized to function as escapements that created limit cycle oscillations at the shoulder's resonant frequency. PMID- 15351888 TI - Crossbow homicides. AB - A total of eight cases of homicide by crossbow are reported, including six intentional, assault-like killings and one hired killer. The bolts showed a high penetration capacity despite the rather low kinetic energy (<100 J): a field-tip traversed one upper arm and the thorax (36 cm) and two broadheads caused perforating injuries of the thorax (25-26 cm). This was due to the high sectional density and the split-like penetration mechanism. Wound morphology was especially important if the perpetrator had extracted the bolt, which occurred in half of the cases. The shape of the entrance wound depended on the type of arrowhead: broadheads produced star-shaped to triangular wounds, field-tips caused circular, oval or slit-like injuries. Foreign material from the arrowhead was found inside two injuries. In assaults, the crossbow was used to hunt the victim down from a short distance which does not require practice but still has the advantage of a distance weapon. However, immediate incapacitation occurred rarely so that additional violence was frequently applied. The noiseless character of the weapon explains why many victims were taken by surprise and why the corpses initially remained unnoticed. Crossbows can therefore be considered ideal weapons for man hunting and some were bought for the very purpose of the killing. PMID- 15351890 TI - Frontotemporal lobar degeneration with motor neuron disease-type inclusions predominates in 76 cases of frontotemporal degeneration. AB - This report presents the largest series of consecutive, neuropathologically confirmed cases of frontotemporal degeneration (FTD). Prior studies have found dementia lacking distinctive histology (DLDH) to be the most common pathology underlying the clinical diagnosis of FTD. In this series of 76 cases, 29 (38%) were found to have frontotemporal lobar degeneration with motor neuron disease type inclusions (FTLD-MND-type) or FTLD-MND (with ALS), the most common neuropathological classification in our series. Only eight (11%) were classified as Pick's disease. Several cases originally designated as DLDH could be reclassified as FTLD-MND-type based on current recommendations for classification of FTD. PMID- 15351889 TI - Chromosomal passengers: the four-dimensional regulation of mitotic events. AB - Chromosomal passengers are proteins that are involved in coordinating the chromosomal and cytoskeletal events of mitosis. The passengers are present in cells as a complex with at least four members: Aurora B, a protein kinase; inner centromeric protein, an activation and targeting subunit; Survivin (function unknown) and Borealin (function also unknown). The kinase is activated at the onset of mitosis, at least partly accomplished by regulation of the levels of its constituents. As mitosis progresses, the kinase complex moves to a highly choreographed series of locations in the mitotic cell, activating key substrates at precise locations and specific times. Functions that require chromosomal passenger activity include chromatin modification (phosphorylation of histone H3), correction of kinetochore attachment errors, aspects of the spindle assembly checkpoint, assembly of a stable bipolar spindle and the completion of cytokinesis. The chromosomal passenger complex provides an essential mechanism for mitotic regulation. PMID- 15351891 TI - A two-hour window for hypothermic modulation of early events that impact delayed opening of the rat blood-brain barrier after ischemia. AB - Opening of the blood-brain barrier (BBB) and consequent edema are known to intensify 24-72 h after ischemic stroke, and research on potential ameliorative therapies in animal models may lead to improved clinical treatments to prevent brain swelling and the secondary damage it causes. In this study, post-ischemic hypothermia treatment, which is an established neuroprotective strategy, was examined for its ability to prevent delayed BBB opening in a rat model of global ischemia. Anesthetized, normothermic SD rats (340-380 g) underwent 20 min of two vessel (carotid) occlusion plus hypotension (2VO ischemia, between 0900-1100 h). Marked cortical BBB leakiness, which developed overnight, was indicated at sacrifice 24 h post-2VO by an average six- to eightfold increase above baseline in transfer constant values (K(i) ) for rate of blood to brain diffusion of intravenously delivered [(3)H]sucrose. A post-2VO treatment involving whole body cooling to 31.5 degrees-32.5 degrees C, maintenance for 6 h and rewarming to normothermia, significantly reduced BBB leakiness at 24 h, whether cooling was initiated immediately after reperfusion, or after a 1-h delay, but not after 2-h delay. Immediate hypothermia treatment reduced overall tissue injury at 24 h as evidenced by an assay of mitochondrial succinate dehydrogenase activity, and also reduced brain edema. By contrast, treatment of rats with the anti-inflammatory drugs cyclosporine A or minocycline offered no protection of BBB or mitochondria. It is concluded that hypothermic alteration of critical events during the first 2 h after prolonged ischemia powerfully mitigates the BBB damage and associated events that normally develop 24 h later. PMID- 15351892 TI - Surgical management of cecal diverticulitis: is diverticulectomy enough? AB - INTRODUCTION: Cecal diverticulitis is a rare condition in the western population. The optimal management of this condition is still controversial, ranging from conservative antibiotic treatment to aggressive resection. We present our experience of the surgical management of eight cases of cecal diverticulitis over a 25-year period. PATIENTS AND METHODS: The mean age of the patients was 54.2 years. Five patients underwent diverticulectomy, 2 patients underwent ileocecal resection, and 1 patient underwent suture of the perforated diverticulum. RESULTS: The postoperative course of all patients was uneventful. At long-term follow-up (mean 14.6 years, range 1-25 years) none of the patients who underwent diverticulectomy, mentioned any symptom or complication. CONCLUSION: We conclude that diverticulectomy, if technically feasible, could be considered as adequate therapy for cecal diverticulitis. Aggressive resection should be considered in cases of extensive inflammatory changes. PMID- 15351893 TI - Day surgery for thyroglossal duct cyst excision: a safe alternative. AB - The goal of this study was to assess which factors affect length of stay in patients operated on for a thyroglossal duct cyst (TDC) and whether day surgery is a safe alternative to a postoperative admission. All charts of patients with TDC excisions at one children's hospital from 1995 to 2001 were reviewed retrospectively. Chi-square tests and multiple logistic regression were used as statistical tests, with p<0.05 considered significant. One hundred children with a mean age of 6 years were operated on for TDC; 46% had day-surgery, with a median length of stay (LOS) of 4 h, and the other 54% were admitted overnight after TDC excision, with a median LOS of 24 h. Factors that significantly prolonged LOS were drain placement, start of surgery after 1 p.m., and surgery being performed by the ENT surgical service rather than the pediatric general surgery service. There was only one hospital readmission for a patient with day surgery at 36 h postoperation for wound hematoma that resolved with nonoperative treatment. Day surgery is safe for routine TDC excisions. There must be a shift in surgeons' behavior if the trend towards an increase in outpatient services for TDC excisions is to continue. PMID- 15351894 TI - Extrahepatic portal hypertension in children: observations on three surgical procedures. AB - This paper presents a comparative prospective study of three modalities of surgical treatment for extrahepatic portal hypertension in children: central splenorenal shunt after splenectomy (CSS), side-to-side lienorenal shunt (SSLR) without splenectomy, and splenectomy and gastroesophageal devascularization (SGD). In an 18-month period, 27 procedures were performed: 10 CSS, 10 SSLR, and seven SGD. The outcomes were evaluated by fall in portal pressures, hematological parameters, shunt patency, splenic regression, and disappearance of esophageal varices. All three procedures were comparable in the fall of portal pressure after surgery. The average blood loss and operating time were statistically significant in favor of SSLR compared with CSS. At 3-month follow-up, shunt patency was confirmed by duplex Doppler study in all the patients in the SSLR group and in nine out of 10 patients in the CSS group. In the CSS and SGD groups, hypersplenism resolved in all the patients. In the SSLR group, blood counts improved in only five out of eight affected children. No patient re-bled during a follow-up of 3-5 years. There were no cases of hepatic encephalopathy or overwhelming postsplenectomy sepsis. In conclusion, CSS is useful when there is a large spleen, severe hypersplenism, and a shuntable splenic vein. SSLR is suitable when there is only mild splenomegaly, mild hypersplenism, and a shuntable splenic vein. Splenectomy and devascularization is the choice when there is no shuntable splenic vein. PMID- 15351895 TI - Surgical closure and reconstruction of a large occipital encephalocele without parenchymal excision. AB - INTRODUCTION: Encephalomeningocele is a herniation of the brain and meninges through a congenital bone defect resulting from the failure of normal midline fusion of the cranial neural tube. Various sites are possible; however, among Caucasians more than 70% of encephaloceles are located on the occipital/posterior part of the cranium. Usually they contain degenerative cerebral cortex, which is often excised during surgical closure of the defect. Techniques to preserve the herniated brain tissue have been described, but they focus on increasing the intracranial cavity by ventriculomegaly and retracting the encephalocele into new intracranial space in a second stage, as a result of hydrocephalic treatment. TECHNICAL REPORT: The authors report a well-documented newborn with a large encephalocystocele of the left occipital lobe. This encephalocele included the left lateral ventricle and a part of the cerebellum, and was covered with allopathic skin. Initially the cystic portion was removed so that a partial reduction of the encephalocystocele was achieved. Six months later, the surgical closure of the defect was performed, with preservation of the occipital and cerebellar parenchyma, by incising the tentorium and retracting the cortex to the newly created infratentorial space. The bony defect was covered with autologous osseous graft harvested from parietal bone and reconstructed. OUTCOME: In the long-term follow-up after 13 years, the child has preserved visual function. PMID- 15351896 TI - Thoracoscopic techniques for the treatment of scoliosis. AB - STUDY DESIGN: To determine the effectiveness of an endoscopic option in an anterior approach to the thoracolumbar spine for scoliosis treatment, 50 patients with follow-ups of 24-45 months were studied retrospectively. OBJECTIVE: The objective was to develop a safe, reproducible, and effective endoscopic technique for the treatment of scoliosis that will provide equal or better outcomes compared with formal open surgical techniques. Techniques for endoscopic treatment of spinal disorders have been under development since 1993. The benefits of thoracoscopic surgery in treating spinal deformities have been documented as improved visualization of the spine, enhanced access to the extremes of the curve, decreased operative times and blood loss, shorter hospital stays and recuperative periods, and decreased overall costs. Following more than 150 endoscopic procedures for the treatment of these spinal deformities, the next progression was to develop a thoracoscopic technique of instrumentation, correction, and fusion for primary thoracic scoliosis. The goals are to gain comparable results, fusion rates, and degrees of correction that meet or exceed the current gold standards of an open procedure. METHODS: From October 1996 to October 1998, 50 patients with a diagnosis of primary thoracic scoliosis were selected to undergo a thoracoscopic technique of instrumentation, correction, and fusion. Postoperatively, patients were assessed for restoration of spinal alignment, axial derotation, pain management, and incidence of complications. RESULTS. Successful endoscopic instrumentation occurred in all patients. Curve correction averaged 50.2%, improving to 68.6% in the last ten cases. Patients with hypokyphosis averaged 20.7 degrees of correction. The preoperative axial rotation, based on the scoliometer, averaged 16 degrees , which was corrected to 5 degrees postoperatively. Postoperative pain was less; patients were off all pain medication by 1-3 weeks compared with patients with a formal open procedure requiring pain medication for 6-12 weeks. The hospital stay averaged 2.9 days. Our initial complication rate was high, which can be attributed to the development of a new technique. The keys to successful fusions included total discectomy, complete endplate removal, and autogenous bone graft. CONCLUSIONS: Although still in early development, the initial results of thoracoscopic techniques are promising. With experience, surgical times are decreasing and fusion and curve correction rates are improving. With further evolution, patients should realize shortened hospitalization, decreased rehabilitation times, and decreased levels of postoperative pain. This is a technically demanding procedure and requires demonstrated skills in endoscopic discectomy and fusion. PMID- 15351897 TI - The orientation of the sandhopper Talitrus saltator during a partial solar eclipse. AB - To acquire more information about the identification and use of the sun and other celestial cues in the sea-land orientation of the sandhopper Talitrus saltator, we carried out releases in a confined environment during a partial solar eclipse and at sunset. The sandhoppers were unable to identify the sun (86% covered) during the eclipse nor to use other celestial compass factors of orientation. This was probably due to the low level of light intensity (close to the minimum level for orientation recorded at sunset) and to the variations in intensity and pattern of skylight polarization. PMID- 15351898 TI - [Screening for diabetic retinopathy and optic disc topography with the "retinal thickness analyzer" (RTA)]. AB - BACKGROUND: The RTA features a fundus photography mode and optic disc topography. Those two new modes were investigated regarding their clinical use. METHODS: First, the combination of wide-field photography with a retinal thickness map was assessed for tele-screening for diabetic retinopathy in 31 eyes. Second, normal values of the optic disc were collected on 30 non-glaucomatous probands in the non-mydriatic mode and compared to the results of the mydriatic mode, for which reproducibility was also investigated. RESULTS: For diabetic retinopathy, the RTA yielded mean 93% sensitivity for proliferative diabetic retinopathy and 100% sensitivity for detecting diabetic macular edema compared to clinical examination at specificities ranging from 58 to 96%. Regarding the measurements of the optic disc, the normal values in mydriasis coincided with those in miosis in all but 4 of 12 parameters. CONCLUSIONS: The RTA is suitable for application in tele screening for diabetic retinopathy. The topography of the optic disc is highly reproducible and may be used for glaucoma diagnostics. PMID- 15351899 TI - Absolute quantification of human liver metabolite concentrations by localized in vivo 31P NMR spectroscopy in diffuse liver disease. AB - Phosphorus-31 NMR spectroscopy using slice selection (DRESS) was used to investigate the absolute concentrations of metabolites in the human liver. Absolute concentrations provide more specific biochemical information compared to spectrum integral ratios. Nine patients with histopathologically proven diffuse liver disease and 12 healthy individuals were examined in a 1.5-T MR scanner (GE Signa LX Echospeed plus). The metabolite concentration quantification procedures included: (1) determination of optimal depth for the in vivo measurements, (2) mapping the detection coil characteristics, (3) calculation of selected slice and liver volume ratios using simple segmentation procedures and (4) spectral analysis in the time domain. The patients had significantly lower concentrations of phosphodiesters (PDE), 6.3+/-3.9 mM, and ATP-beta, 3.6+/-1.1 mM, (P<0.05) compared with the control group (10.0+/-4.2 mM and 4.2+/-0.3 mM, respectively). The concentrations of phosphomonoesters (PME) were higher in the patient group, although this was not significant. Constructing an anabolic charge (AC) based on absolute concentrations, [PME]/([PME] + [PDE]), the patients had a significantly larger AC than the control subjects, 0.29 vs. 0.16 (P<0.005). Absolute concentration measurements of phosphorus metabolites in the liver are feasible using a slice selective sequence, and the technique demonstrates significant differences between patients and healthy subjects. PMID- 15351900 TI - Direct magnetic resonance arthrography. AB - Magnetic resonance (MR) arthrography has gained increasing popularity as a diagnostic tool in the assessment of intra-articular derangements. Its role has been studied extensively in the shoulder, but it also has been explored in the hip, elbow, knee, wrist and ankle. This article reviews the current role of direct MR arthrography in several major joints, with consideration of pertinent anatomy, techniques and applications. PMID- 15351901 TI - MRI in coronary artery disease. AB - Diagnosis of coronary artery disease (CAD) is a major challenge for medical imaging, because CAD is the leading cause of death in developed nations. Several non-invasive tests are used in clinical routine for the detection of CAD. However, due to limited sensitivity and specificity, the reliable diagnosis as well as the exclusion of CAD can only be established by catheter angiography. In patients with known CAD, therapeutic decisions require accurate information on myocardial function, ischemia and viability. Recently, magnetic resonance (MR) imaging has emerged as a non-invasive cardiac imaging technique that provides information on cardiac morphology, cardiac function, myocardial viability, and coronary morphology. This review discusses technical aspects and the clinical impact of different MR techniques. PMID- 15351902 TI - MR-guided transgluteal biopsies with an open low-field system in patients with clinically suspected prostate cancer: technique and preliminary results. AB - The purpose of this study was to examine the feasibility and safety of MR-guided biopsies with a transgluteal approach in patients with uncertain or suspicious prostate lesions. Twenty-five patients with uncertain or suspicious focal prostate lesions detected by high-field MR imaging of the prostate gland using endorectal coil imaging were biopsied with a transgluteal approach in a low-field MRI system (0.2 T, Concerto, Siemens). The procedures were guided using T1 weighted FLASH sequences. The prostate gland was biopsied repeatedly with a coaxial technique through a 15-gauge pencil tip with a 16-gauge biopsy handy (median 3.8 samples per patient). Complications and biopsy findings were documented retrospectively. Using T1-weighted sequences biopsy procedures were performed successfully with MR guidance in all cases without any side effects or complications. The median intervention time was 11.3 min. Pathological findings revealed ten cases of hyperplasia or atrophy, three cases of prostatitis, ten cases of carcinoma and two cases of normal tissue. The clinical follow-up showed that in two patients prostate cancer was missed at MR-guided biopsy. Transgluteal MR-guided biopsy of the prostate gland is a safe and promising approach for histological clarification of uncertain or suspicious lesions. PMID- 15351903 TI - Radiation-induced temporary hair loss as a radiation damage only occurring in patients who had the combination of MDCT and DSA. AB - As imaging technologies become increasingly advanced, it is possible to obtain detailed morphological information as well as functional imaging data. In some imaging technologies, the radiation dose increases with the ability to obtain better images or more detailed information. We encountered three cases of temporary bandage-shaped hair loss, which was caused by perfusion studies of the head by multi-detector row computed tomography (MDCT) for evaluation of cerebral blood flow in patients with vascular disorders. In all three patients with temporary hair loss, two angiographies of the head had been performed in the period of serial CT examinations. This suggested the possibility that radiation exposure from angiography performed in serial examinations, combined with the perfusion studies of the head with MDCT, played an important role in this temporary, bandage-shaped hair loss. Radiologists should be aware that a cumulative or multiplier effect of radiation exposure from multiple diagnostic techniques may result in hair loss and other types of radiation complications. PMID- 15351904 TI - MR demonstration of septal involvement in arrhythmogenic right ventricular dysplasia. AB - Arrhythmogenic right ventricular dysplasia (ARVD) is a heart muscle disease of unknown origin. Although MR imaging is regarded as the best technique for the demonstration of functional and structural abnormalities in ARVD, fat deposits in the interventricular septum have never been documented on MR imaging. We report the case of interventricular septal fatty deposition demonstrated by fat suppressed MR imaging in a 48-year-old man. PMID- 15351905 TI - Compression of the extrahepatic bile duct by the portal vein: a diagnostic pitfall of MR cholangiopancreatography. PMID- 15351906 TI - Specificity of choline metabolites for in vivo diagnosis of breast cancer using 1H MRS at 1.5 T. AB - The purpose was to determine if in vivo proton magnetic resonance spectroscopy ((1)H MRS) at 1.5 T can accurately provide the correct pathology of breast disease. Forty-three asymptomatic volunteers including three lactating mothers were examined and compared with 21 breast cancer patients. Examinations were undertaken at 1.5 T using a purpose-built transmit-receive single breast coil. Single voxel spectroscopy was undertaken using echo times of 135 and 350 ms. The broad composite resonance at 3.2 ppm, which includes contributions from choline, phosphocholine (PC), glycerophosphocholine (GPC), myo-inositol and taurine, was found not to be a unique marker for malignancy providing a diagnostic sensitivity and specificity of 80.0 and 86.0%, respectively. This was due to three of the asymptomatic volunteers and all of the lactating mothers also generating the broad composite resonance at 3.2 ppm. Optimised post-acquisitional processing of the spectra resolved a resonance at 3.22 ppm, consistent with PC, in patients with cancer. In contrast the spectra recorded for three false-positive volunteers, and the three lactating mothers had a resonance centred at 3.28 ppm (possibly taurine, myo-inositol or GPC). This improved the specificity of the test to 100%. Careful referencing of the spectra and post-acquisitional processing intended to optimise spectral resolution of in vivo MR proton spectra from human breast tissue resolves the composite choline resonance. This allows the distinction of patients with malignant disease from volunteers with a sensitivity of 80% and specificity of 100%. Therefore, resolution of the composite choline resonance into its constituent components improves the specificity of the in vivo (1)H MRS method, but does not overcome the problem of 20% false-negatives. PMID- 15351907 TI - Rotations of three-joint fingers: a radiological study. AB - The aim of the current study was to test a protocol of quantification of phalangeal three-dimensional (3D) rotations during flexion of three-joint digits. Three-dimensional-specific software was developed to analyze CT reconstruction images. A protocol was carried out with six fresh-frozen upper limbs from human cadavers free from any visible pathology (three females, three males). CT millimetric slices were done for reconstruction of hand bone units. Orthonormal coordinate systems of inertia were calculated for each unit. Three-dimensional phalangeal rotations were estimated between two static positions (fingers in extension and in a fist position). Results were displayed for the joints of each three-joint finger with calculation of 3D rotations. Mean longitudinal axial rotations of metacarpophalangeal (MCP), proximal interphalangeal (PIP) and distal interphalangeal (DIP) joints ranged from 14 degrees pronation to 19 degrees supination. The index finger was in a global pronation position (4/6 specimens). The fourth and fifth fingers were in a global supination position in every case. The third finger was in a more variable global rotation (pronation in 2/6 specimens). MCP, PIP and DIP flexion angles ranged respectively from 71 degrees to 89 degrees, 65 degrees to 87 degrees, and 41degrees to 77 degrees. Lateral angles ranged from 19 degrees (ulnar angulation) to 23 degrees (radial angulation). The study of phalangeal rotations was possible in spite of a heavy protocol. This protocol could be partially automatated to speed up the analyses. Longitudinal axial rotations could be analyzed, in addition to flexion/extension or abduction/adduction rotations. CT scan reconstructions would be helpful for investigating pathological fingers. Abnormal rotations of digits could be quantified more precisely than during a current clinical examination of the hand. PMID- 15351912 TI - Decreased uptake after fractionated ablative doses of iodine-131. AB - PURPOSE: In an attempt to obviate the necessity for hospitalisation, the ablative dose of 131I in the treatment of thyroid cancer is divided into two or three fractions at weekly intervals in some hospitals with no special bed for 131I treatment. Thyroid stunning has been observed in patients receiving a 131I dose between 74 and 370 MBq (2-10 mCi). However, the influence of 131I uptake after administration of a higher dose, such as 1,110-1,850 MBq of 131I, has never been reported. In this study, we evaluated the degree of reduction in 131I uptake after patients received 1,480 MBq of 131I and evaluated the clinical value of fractionated ablative doses of 131I. METHODS: Thirty-five patients with functional thyroid cancer received a total of 4,440 MBq (120 mCi) of 131I which was divided into three fractions administered at weekly intervals. In all patients two 131I whole-body scans were performed. The first scan was performed directly prior to the second dose of 131I (7 days after the first administration of 131I), and the second scan was performed 7 days after the second administration of 131I and directly prior to the third administration. Regions of interest including the neck and lungs were drawn to calculate the uptake of 131I in the thyroid remnant and possible cervical lymph node and lung metastases. RESULTS: The mean uptake of 131I was 2.73% 7 days after the first administration, and decreased significantly to 0.26% 7 days after the second administration. The mean decrease was as high as 80.7%. The decrease in 131I uptake was significant in all patients except the two with lung metastases. In the two patients with lung metastases, no definite evidence of decreased uptake was noted; the uptake of 131I in the lung metastases even increased on the second 131I image in one of these patients. After administration of 1,480 MBq of 131I, the decreased uptake was significant in all neck lesions but not in lung metastases. CONCLUSION: The use of fractionated ablative doses of 131I is not to be recommended in patients without lung metastases. However, the influence of fractionated ablative doses of 131I in patients with lung metastases is worthy of further study. PMID- 15351913 TI - Precise localisation of a sentinel lymph node in a rare drainage region with SPECT/MRI using interstitial injection of 99mTc-nanocolloid and superparamagnetic iron oxide. PMID- 15351914 TI - From the magic bullet to an effective therapy: the peptide experience. PMID- 15351916 TI - Chondroid lipoma: correlation of imaging findings and histopathology of an unusual benign lesion. AB - The imaging findings of soft tissue tumours are often non-specific and generally require biopsy to differentiate between benign and malignant lesions. The finding of curvilinear, annular or amorphous mineralisation in an enlarging mass has sinister connotations. In this case report, we present the imaging findings with histological correlation of a chondroid lipoma, an unusual benign soft tissue tumour, which presented with radiographic evidence of calcification, an imaging finding not previously described. We also describe the ultrasound appearance and certain MR imaging appearances that have not been previously attributed to this tumour in the few reported cases. PMID- 15351918 TI - Establishment of a gene transfer system for Rhodothermus marinus. AB - Genetic manipulation of Rhodothermus marinus has been hampered by the lack of a selection system for gene transfer. We report construction of a Rhodothermus/Escherichia coli shuttle plasmid, containing the R. marinus trpB gene, based on pUC18 and the cryptic R. marinus plasmid pRM21. A plasmid-less R. marinus recipient strain was selected on the basis of growth characteristics and absence of restriction activity. The shuttle plasmid, pRM100, was successfully introduced into a TrpB- mutant of the recipient strain using electroporation and was found to transform it to prototrophy. No loss or rearrangement of pRM100 was observed after growth for 80 generations in non-selective medium. The relative copy numbers of pRM100 and of the parental plasmid, pRM21, were determined as 7+/ 1 and 42+/-4, respectively. The shuttle plasmid was used to optimize an electroporation protocol, and the maximal number of transformants obtained was 4.3+/8-0.7x10(6) cfu/microg DNA at 22.5 kV/cm, 200 Omega and 25 microF. Transformation failed, however, after chemical preparation of cells according to several protocols. This is the first report of genetic transformation in the genus Rhodothermus. PMID- 15351919 TI - Evolution of MHC-DRB class II polymorphism in the genus Apodemus and a comparison of DRB sequences within the family Muridae (Mammalia: Rodentia). AB - Allelic diversity at major histocompatibility complex (MHC) genes is thought to be maintained by balancing selection over long periods of time, even across multiple speciation events. Trans-species sharing of MHC alleles among genera has been supported by many studies on mammals and fish, but in rodents, the results are ambiguous. We investigated natural levels of MHC- DRB variability and evolutionary processes in the wood mouse (Apodemus sylvaticus) and the yellow necked mouse (Apodemus flavicollis), which are common, sympatric murid rodents in European forests. Using single-strand conformation polymorphism analysis and DNA sequencing, 38 DRB exon 2 alleles were detected among 162 A. sylvaticus from nine different locations in Germany and Switzerland, and 15 DRB exon 2 alleles were detected among 60 A. flavicollis from three different locations in northern Germany. There was evidence for balancing selection in both species. Phylogenetic analysis, including additional murid taxa, showed that the DRB exon 2 sequences did not separate according to species, consistent with trans-species evolution of the MHC in these taxa. PMID- 15351920 TI - Endobronchial carcinoid in a child: depiction with three-dimensional volume rendering. AB - Endobronchial carcinoid is an extremely rare neoplasm in children. We report a case in which this entity was prospectively diagnosed by 3-D CT bronchography with volume rendering techniques. The ability to prospectively implicate an endobronchial neoplasm in a child may help to guide subsequent therapy and management. PMID- 15351921 TI - Tailgut cyst in a child. AB - Tailgut cyst, or retrorectal cystic hamartoma, is a rare congenital lesion found in the presacral space. The lesion has been infrequently reported in the literature. We report the MRI findings of a tailgut cyst in a 2-year-old girl who presented with a sacral dimple and skin discoloration. PMID- 15351922 TI - Primary bacterial peritonitis in otherwise healthy children: imaging findings. AB - We report the imaging findings of two recent cases of primary bacterial peritonitis in otherwise healthy children with a clinical presentation mimicking acute appendicitis. Primary bacterial peritonitis is rare in the absence of underlying systemic disease. Although it has been described in the pediatric literature, the imaging findings have not been described in the radiological literature to the best of our knowledge. With imaging playing an increasing role in the evaluation of appendicitis in children, it is important for the radiologist to be familiar with this inflammatory process. PMID- 15351923 TI - Premedication medicines do not cause drug metabolic interaction with propofol using human liver microsomes in vitro. AB - OBJECTIVE: Propofol (2,6-diisopropylphenol) is widely used for anesthetic induction as well as for chronic sedation in intensive care units. In this study, we investigated the interaction between propofol and premedications, i.e., psychotropic and antianxiety agents (diazepam, midazolam), hypnotics (thiamylal), local anesthetics (lidocaine), depolarizing muscular relaxants (vecuronium), an antihypertensive (clonidine) and an H2-receptor antagonist (cimetidine) using human liver microsomes in vitro. METHODS: The interaction effects between propofol and premedications were examined using human liver microsomal preparation in vitro. The concentration of propofol was determined by HPLC with UV detection. RESULTS: The apparent Michaelis-Menten constant (Km) and the maximal velocity of total metabolic formation (Vmax) of propofol in human liver microsomes were 123 microM and 26.1 micromol/min per milligram of mg protein, respectively. Seven premedications (diazepam, midazolam, thiamylal, lidocaine, cimetidine, vecuronium, and clonidine) did not inhibit propofol metabolism in human liver microsomes at concentrations within the therapeutic range. CONCLUSIONS: These results showed no interactions between propofol and seven premedication drugs within the therapeutic range of propofol using human liver microsomes in vitro. PMID- 15351924 TI - Requirements for a successful implementation of drug interaction information systems in general practice: results of a questionnaire survey in Germany. AB - OBJECTIVE: To determine drug interaction information requirements in general practice with respect to both content and mode of presentation. METHODS: In a mail survey among 2,000 general practitioners in south-west Germany, we collected information on risk evaluation of drug interactions and combinations of concern, usage of and satisfaction with the current information sources, desirable content and mode of future presentation and demographic variables. Categorical variables were compared using chi2 test. Trends were analysed with Cochran-Armitage test and determinants of literature usage with logistic regression. RESULTS: Response rate was 60.8%. The majority of general practitioners considered drug interactions a risk factor in prescribing (88.6%). For 18.2% of the drug combinations most frequently indicated as interacting, there was no published evidence of a clinically relevant interaction. More than half of the participants were dissatisfied with the information on severity, mechanism, and dose adjustment currently available in their sources. In particular, non-interacting alternatives were thought to be lacking (86.9%). Users of drug interaction software more frequently retrieved drug interaction information than non-users [odds ratio (OR) 1.95; 95% confidence intervals (CI) 1.50, 2.52], but only 28.6% of general practitioners had access to such systems. There was a significant trend towards electronic sources among younger physicians, but at present, 41.7% of general practitioners favour printed sources, and 8.8% would refuse to use electronic sources. CONCLUSION: General practitioners wish for more informative support on drug interactions, especially concerning management. Despite a trend towards electronic information sources, printed documents are presently still required to reach all prescribers. PMID- 15351925 TI - Reinnervation of the rat musculocutaneous nerve stump after its direct reconnection with the C5 spinal cord segment by the nerve graft following avulsion of the ventral spinal roots: a comparison of intrathecal administration of brain-derived neurotrophic factor and Cerebrolysin. AB - Experimental model based on the C5 ventral root avulsion was used to evaluate the efficacy of brain-derived neurotrophic factor (BDNF) and Cerebrolysin treatment on motor neuron maintenance and survival resulted in the functional reinnervation of the nerve stump. In contrast to vehicle, BDNF treatment reduced the loss and atrophy of motor neurons and enhanced the regrowth axon sprouts into the distal stump of musculocutaneous nerve. However, the axon diameter of the myelinated fibers was smaller than those of control rats. The morphometric results were related to a low score in behavioral test similar to vehicle-treated rats. Cerebrolysin treatment greatly protected the motor neurons against cell death. Moreover, morphometric features of myelinated axons were better than those of rats treated with vehicle or BDNF. The mean score of grooming test suggested better results of the functional motor reinnervation than after BDNF administration. The majority of rescued motor neurons regenerating their axons through nerve graft in both BDNF- and Cerebrolysin-treated rats expressed choline acetyltransferase immunostaining. The results demonstrate that BDNF has more modest effects in preventing the death of motor neurons and functional recovery of injured motor nerve after root avulsion than Cerebrolysin. PMID- 15351926 TI - Perceived reachability: the roles of handedness and hemifield. AB - The impact of reaching experience on the ability to predict one's own reaching range was investigated. Left- and right-handed participants made verbal estimates about the reachability of a target object for both arms in ipsilateral and contralateral frontal space. There was a significant overestimation bias in both groups and for both hands. The overestimation bias increased with the target object's eccentricity in contralateral space. The implications of these findings for models of motor control and motor imagery are discussed. PMID- 15351927 TI - Excitability changes in human hand motor area dependent on afferent inputs induced by different motor tasks. AB - Using the technique of transcranial magnetic stimulation (TMS) with a figure-of eight-shaped coil in 16 normal volunteers, we studied the extents of motor evoked potentials (MEPs) induced by remote facilitation of voluntary teeth clenching (VTC) and by motor imagery (MI). In particular, we examined whether different excitability changes in the primary motor cortex (M1) induced by both facilitation methods occur between early (I1 and I2) and late (I3 and I4) components of I-waves elicited from a first dorsal interosseous (FDI) muscle. Both components of I-waves were induced by anterior-medially (AM) directed currents or posterior-laterally (PL) directed currents. Our hypothesis was that facilitatory effects of VTC and MI on M1 differ because the neural pathways of these afferent inputs differ. The present results indicate that during MI MEP amplitudes of late components are significantly larger than those of early ones, although both MEP amplitudes are enhanced. On the other hand, during VTC MEP amplitudes of early components are significantly enhanced, but those of late ones are rather depressed. We conclude that recruitment of early and late components of I-waves differ depending on the afferent inputs to the motor cortex. PMID- 15351928 TI - Nociceptin inhibits vanilloid TRPV-1-mediated neurosensitization induced by fenoterol in human isolated bronchi. AB - Chronic exposure to beta(2)-adrenoceptor agonists, especially fenoterol, has been shown to increase smooth muscle contraction to endothelin-1 in human bronchi partly through tachykinin-mediated pathways. The purpose of this work was to further investigate the role of sensory nerves in fenoterol-induced sensitization of human airways and the effect of nociceptin, a nociceptin/orphanin FQ (NOP) receptor agonist, on the increase in contraction after fenoterol exposure. Human bronchi from 62 patients were sensitized to endothelin-1 by prolonged incubation with fenoterol (0.1 microM, 15 h). The sensitizing effect of fenoterol was inhibited by high concentration of capsaicin (10 microM, 30 min before fenoterol sensitization), which induces depletion of mediators from sensory nerves, or co incubation of fenoterol and capsazepine (1 microM), a vanilloid TRPV-1 receptor antagonist. Moreover, short pretreatment of bronchi with capsaicin (10 microM) or capsazepine (1 microM) after sensitization by fenoterol decreased the rise in smooth muscle contraction to endothelin-1. Nociceptin (1 microM) also inhibited the increased contraction in fenoterol-sensitized bronchi. Tertiapin (10 microM), an inhibitor of the inward-rectifier K(+) channels, but not naloxone (0.1 microM), a DOP/KOP/MOP receptor antagonist, prevented the inhibitory effect of nociceptin. In conclusion, fenoterol induces sensitization of human isolated bronchi to endothelin-1 in part through the stimulation of the vanilloid TRPV-1 receptor on tachykininergic sensory nerves. Nociceptin inhibits airway hyperresponsiveness via NOP receptor activation. This effect involves inward rectifier K(+) channels. PMID- 15351929 TI - Robust simple sequence repeat markers for spruce (Picea spp.) from expressed sequence tags. AB - Traditionally, simple sequence repeat (SSR)markers have been developed from libraries of genomic DNA. However, the large, repetitive nature of conifer genomes makes development of robust, single-copy SSR markers from genomic DNA difficult. Expressed sequence tags (ESTs), or sequences of messenger RNA, offer the opportunity to exploit single, low-copy, conserved sequence motifs for SSR development. From a 20,275-unigene spruce EST set, we identified 44 candidate EST SSR markers. Of these, 25 amplified and were polymorphic in white, Sitka, and black spruce; 20 amplified in all 3 spruce species tested; the remaining five amplified in all except one species. In addition, 101 previously described spruce SSRs (mostly developed from genomic DNA), were tested. Of these, 17 amplified across white,Sitka, and black spruce. The 25 EST-SSRs had approximately 9% less heterozygosity than the 17 genomic-derived SSRs (mean H=0.65 vs 0.72), but appeared to have less null alleles, as evidenced by much lower apparent inbreeding (mean F=0.046 vs 0.126). These robust SSRs are of particular use in comparative studies,and as the EST-SSRs are within the expressed portion of the genome, they are more likely to be associated with a particular gene of interest, improving their utility for quantitative trait loci mapping and allowing detection of selective sweeps at specific genes. PMID- 15351930 TI - [Whole-body MRI in comparison to skeletal scintigraphy for detection of skeletal metastases in patients with solid tumors]. AB - PURPOSE: The aim of this study was to compare the diagnostic efficacy of whole body magnetic resonance imaging (WB-MRI) as a new and rapid examination technique with skeletal scintigraphy for detection of skeletal metastases from solid tumors. MATERIAL AND METHODS: In 129 patients with solid malignant tumors, WB-MRI was performed for individual comparison with skeletal scintigraphy. Examinations were performed with the innovative AngioSURF rolling table with integrated phased array surface coil and coronary TIRM sequences for different body regions. RESULTS: The results for WB-MRI and skeletal scintigraphy were concordant in 81% of the cases, whereby both procedures excluded skeletal metastases in 43%. WB-MRI and skeletal scintigraphy demonstrated skeletal metastases in 38% of the cases, whereby WB-MRI provided more comprehensive findings in 45%. In 12% of the cases, skeletal scintigraphy was superior to WB-MRI and in 19% the findings were discordant, whereby WB-MRI detected skeletal metastases in 15 cases which had not been found on skeletal scintigraphy. In nine cases, skeletal scintigraphy was positive when the WB-MRI was negative. In 60% of the cases, WB-MRI evidenced tumor-associated findings. CONCLUSION: WB-MRI represents a promising new staging technique for detection of skeletal metastases, which is more sensitive in many cases than skeletal scintigraphy in detecting and assessing the extent of skeletal metastases-and tumor-associated findings that are relevant for treatment strategy. PMID- 15351931 TI - [Right ventricle in arterial hypertension]. AB - Left ventricular hypertrophy is the best known and most evident cardiac organ manifestation of arterial hypertension. However, only limited findings are available on function and structure of the right ventricle. The published studies show that the right ventricle is also affected in the course of hypertensive heart disease. Particularly hypertrophy of the right ventricular free wall, impaired right ventricular diastolic filling, elevated right ventricular filling pressure as well as impairment of right ventricular ejection fraction in the late phase of arterial hypertension were reported. Possible causes of these structural and functional changes of the right ventricle are (1) translation of the increased left ventricular filling pressure in the pulmonary circulation, (2) interaction of the right and left ventricle, and (3) systemic circulation of cytokines and growth hormones such as angiotensin II, aldosterone, and others. PMID- 15351932 TI - Diagnostic significance of gamma-interferon in tuberculous pleurisy. AB - We studied ADA and gamma interferon (gamma-IFN) levels in pleural fluid of 45 cases presenting with pleural effusion to the Ankara University School of Medicine Chest Diseases Hospital between September 2001 and September 2002. Fifteen patients had TB pleurisy, 20 patients had malignant pleurisy and 10 patients had transudative pleural effusion. The cut-off value for pleural fluid gamma-IFN levels were 12 pg/mL. According to this, all patients with transudative effusions, 19 of 10 patients with malignant effusions and 2 of 15 patients with tuberculous (TB) effusions had pleural fluid gamma-IFN levels under the cut-off value. In exudative effusions, sensitivity and specificity of gamma-IFN were 87% and 95% respectively. The sensitivity of pleural fluid ADA levels was 86% and specificity of pleural fluid ADA levels was 100%. Pleural fluid ADA levels in TB effusions were significantly higher than the non-TB effusions. Also there were no statistically significant differences between pleural fluid ADA and g-IFN levels according to sensitivity and specificity. As a result, we have shown that gamma IFN is a valuable test in diagnosis of TB pleurisy. We think that when it is used routinely, it will be a good alternative to the conventional invasive diagnostic tests. PMID- 15351933 TI - [The nitrosative effect of peripheral ischemia-reperfusion on lung and preventive of caffeic acid phenethyl ester]. AB - Ischemia-reperfusion (IR) is characterized by microvascular disfunction and this involves both direct effected organ and remote organ by systemic inflammatory response. These remote effects of IR are most frequently observed in the lung and cardiovascular system. In this study we aim to determine lung damage which induced IR, and endothelial and microvascular disfunction using nitrosative markers. Previous studies suggest that caffeic acid phenethyl ester (CAPE) has some antioxidant effects. Therefore, we also investigated whether it has a role associated with nitric oxide during IR condition. Twenty-two adult male Wistar rats were divided into three groups: control (n= 7), IR (n= 7), and CAPE + IR (n= 8). 8 h IR period was performed on right hindlimb in the IR and the CAPE with IR group. In the CAPE with IR group, animals received CAPE 10 microM 1 h before the reperfusion. At the end of the reperfusion period, blood, bronchoalveolar lavage (BAL) and lung tissue were obtained, and were used for biochemical and histopathological examination. There was a significantly elevation in serum nitrate, BAL MPO, and leukocyte infiltration in the lung in the IR group compared to the CAPE + IR group. But, serum nitrite and lung 3-NT levels were not different between these groups. While nitrate (p< 0.0001), MPO (p< 0.0001) and leukocyte infiltration (chi2= 27.163, p= 0.0001), reduce by using CAPE before reperfusion, tissue 3-NT levels did not change. In conclusion, peripheral IR leads to systemic inflammatory responses and endothelial disfunction-induced NO production, and these harmful effects may reduced by CAPE. PMID- 15351934 TI - Adaptation of functional outcomes of sleep questionnaire (FOSQ) to Turkish population. AB - Turkish version of functional outcomes of sleep questionnaire (FOSQ.tr) was examined for its psychometric properties in patients with obstructive sleep apnoea hypopnoea syndrome (OSAHS). The FOSQ was translated into Turkish using a forward-backward translation. For the psychometric evaluation, 73 consecutive patients were selected along with 73 control subjects. Internal consistency, test retest reliability, concurrent and discriminant validity were investigated. Values of Cronbach's alpha for the total FOSQ.tr (alpha= 0.92) and its sub-scales (alpha= 0.76-0.80) suggest that the questionnaire was consistent internally. Test retest reliability of the FOSQ.tr was significant for the total score (r= 0.7) and the sub-scales apart from social outcome (r= 0.5 to 0.8, all p< 0.01). FOSQ.tr correlated moderately with Epworth sleepiness scale (ESS), with coefficients ranging from r= -0.5 to -0.62, (all p< 0.05) for the sub-scales, and r= -0.64 (p< 0.01) for total score. Discriminant analysis showed that FOSQ.tr could significantly distinguish the patients from normal subjects (p< 0.03). The psychometric properties of the FOSQ.tr suggest that it is a valid and reliable instrument for the assessment of the impact of disorders of excessive sleepiness on daily behaviour. PMID- 15351935 TI - [Determination of environmental tobacco smoke in primary school children with urine cotinine measurements]. AB - Environmental tobacco smoke (ETS) has been regarded as one of the most important public health issues. It has been estimated that approximately 75% of Turkish children are exposed to ETS. In this study the parental smoking habits were determined. Then, the relationship between parent-reported estimates of children's exposure to ETS in the home and children's urinary cotinine levels was examined. According to the reports of parents, 57.8% of the fathers and 23.3% of the mothers were current smokers, 69.8% of the children came from homes with smokers, and 53.4% had daily exposure to ETS. Urinary cotinine levels were significantly higher in the exposed group than the nonexposed group. This data showed that ETS exposure was prevalent and a combination of a parent-report and a biological measures is suggested as the most informative estimate of ETS exposure in children. PMID- 15351936 TI - [The effects of hormone replacement therapy on pulmonary functions in postmenopausal women]. AB - Although the effects of hormone replacement therapy (HRT) used after the menopause on the cardiovascular system, bone tissue, mammary gland, endometrium, menopausal symptoms and sexuality has been well known, there are very few studies about the effects of HRT on the pulmonary functions. In this study, we evaluated the effects of HRT on the pulmonary functions. For this purpose the postmenopausal women were randomized into two groups, 0.625 mg conjugated equine estrogens + 5 mg medroxyprogesterone acetate as a HRT was given to the group I (n= 32), and placebo was given to the control group. Pulmonary function tests (PFT) were performed to the all cases by same technician before and three months after the treatment. FVC, FEV1, FEV1/FVC, FEF25-75 and PEF parameters of the groups were compared. There were no statistically significant differences between the two groups considering the age, duration of the menopause and body mass index (BMI) (p> 0.05). There were no statistically significant differences between the groups considering FVC, FEV1, FEV1/FVC and FEF25-75 values of initial and the 3rd months measurements (p> 0.05). But PEF levels of the HRT group were significantly different than the initial values three months after the treatment (4.42 +/- 2.6 and 4.84 +/- 1.1) (p= 0.023). There were no statistically significant differences between the other compared parameters of PFT. As a result among the PFT parameters of the postmenopausal women taking HRT, PEF that was used as on obstruction parameter was found elevated. So, we concluded that more detailed and prospective studies were needed about the effects of HRT on PFT. PMID- 15351937 TI - Effects of antithyroid medication on the flow-volume loop in patients with hyperthyroidism. AB - This prospective study was designed to evaluate the effects of hyperthyroidism on flow-volume loops in nonasthmatic 20 patients with hyperthyroidism. Thyroid related hormones (Total T3, Total T4 and TSH), thyroid gland volumes with ultrasonography, circumference of neck values and flow-volume loops were obtained at the beginning and after three months of antithyroid treatment. Propylthiouracil treatment was followed by a statistically significant decrease in thyroid gland volume and circumference of neck (p< 0.001 and p< 0.001, respectively). The most significant result was improvement of maximum midexpiratory flow rate (MMEFR) after propylthiouracil therapy for three months (p= 0.003). Increases in mean forced expiratory flow after 25% of FVC has been exhaled (FEF25), mean forced expiratory flow after 75% of FVC has been exhaled (FEF75) values were found consistent with the overall improvement in expiratory flow parameters (p= 0.044, p= 0.012 respectively). In conclusion, we speculated that improvement of expiratory flow parameters might be the earlier changes in flow volume loops of patients who were treated with propylthiouracil for hyperthyroidism. PMID- 15351938 TI - Risk factors associated with postoperative pulmonary complications following oncological surgery. AB - The purpose of our study was to determine the incidence of different postoperative pulmonary complications (PPCs) and their associated risk factors in patients who have undergone various elective surgical procedures in an oncological surgery center. Ninety five adult patients were studied prospectively for one year period. For the study group, predictors of pulmonary complications of interest were determined as age, gender, body mass index, co morbid conditions (preexisting history of chronic obstructive pulmonary disease, asthma, bronchiectasis, restrictive lung disease), site and type of the operation, smoking history, The American Society of Anesthesiologists (ASA) physical status, physical examination and chest X- Ray findings, pulmonary function tests, type and duration of anesthesia, surgical incision site and length and presence of nasogastric tube suction. The PPC rate of our study group was 40% (38/95). Atelectasis and bronchospasm were the most frequently observed PPCs (13.7%) Among all the risk factors taken into consideration, only three were found to be significant independent predictors of pulmonary complications according to multivariate analysis as follows: incision location concerning abdomen (p= 0.008), duration of anesthesia per hour (p= 0.0001), values of FEV1 < 50% (p= 0.007). Our data revealed that the incidence of PPCs was high in our study group when compared to results of general population. Application of major resection surgeries for cancer patients can be an explanation for this result. Shortening the duration of surgery, avoiding general anaesthesia in selected group of patients may reduce the risk of PPCs. PMID- 15351939 TI - [Body mass index and serum and sputum TNF-alpha levels relation in asthma and COPD]. AB - Previous studies have revealed the relationship between asthma with obesity and low body mass index (BMI) with chronic obstructive pulmonary disease (COPD). Tumor necrosis factor-alpha (TNF-alpha) is thought to be related with low BMI. The aim of this study was to determine sputum and serum TNF-alpha levels in patients with COPD and asthma and to evaluate whether these parameters had correlation with BMI. Thirty patients with moderate persistent asthma and 26 patients with moderate -severe COPD were included. After BMI values were calculated, sputum was induced by inhalation of hypertonic saline solution and blood was drawn for analysis of serum TNF-alpha levels. There were significant differences in age, serum and sputum TNF-alpha levels between asthma and COPD subjects (Sputum TNF-alpha: asthma; 513 +/- 151 pg/mL-COPD: 333 +/- 126 pg/mL, p< 0.001; Serum TNF-alpha: asthma; 332 +/- 114 pg/mL-COPD: 197 +/- 81 pg/mL, p< 0.001), however there was no difference in BMI (asthma; 28 +/- 5.7-COPD; 26.6 +/- 12.9, p= 0.1). Patients were divided into four categories according to their BMI values as underweight, normal, overweight and obese. In asthmatics; there were 12 (40%) obese and 11 (36%) overweight patients while 9 (34%) of COPD patients were underweight. No significant difference was observed among these four groups according to serum-sputum TNF-alpha and smoking history both in asthmatics and in COPD subjects. While there was no correlation between BMI and serum-sputum TNF alpha levels, BMI was significantly correlated with both smoking history and duration of disease in COPD patients. As a result, most of the asthmatic patients were described as overweight and obese while no such variation was noted in the COPD patients. The induced sputum TNF-alpha levels has no additional benefit on serum TNF-alpha levels which has already known to be associated with BMI. PMID- 15351940 TI - [Synchronous primary lung cancers: due to the four cases]. AB - If two primary lung cancers are present in the same time, it diagnosed as synchronous lung cancer. It constitutes 0.1-1.6% of all lung cancers. In this report, we described four cases diagnosed synchronous multiple primary lung cancers (MPLC) in between June 1999 and May 2002. Mean age was 63, and all of those were male. All patients were heavy smoker (mean 58 packet/year). Histology of lung cancers was squamous-adeno in two cases, squamous-small cell lung cancer in one case, and squamous-squamous in two cases. All patients had bilateral mass lesions and no mediastinal and systemic spread. Lesion of second primary lung cancer was unable to seen in two patient's chest X-ray. Their lesions were seen by computerized tomography in one case, and by bronchoscope only in the other case. Lesion site was left lower lobe, right lower lobe, right middle lobe, and left upper lobe (25% for each). Diagnosis of lung cancer was made by transthoracic needle biopsy in three lesion, and made by bronchoscopic biopsy in other lesions. While it is recommended that, all of MPLC lesions were staged separately and treated surgically, due to the advanced age, presence of small cell lung cancer, and inadequate post-operative respiratory reserve, surgical treatment could not apply to these patients and they received chemotherapy. Three of them was died (mean survival was 11 months). Due to the this report, we emphasized that diagnostic procedures should be done separately for each lesions in patients who had more than one lesion, and treatment should plane according to these results. PMID- 15351941 TI - Tracheobronchopathia osteochondroplastica: two cases and review of literature. AB - Tracheobronchopathia osteochondroplastica (TO) is an unusual disorder. It mainly affects men over 50 years old and clinical manifestations are observed when obstructive or infectious complications occur. A 50-year old woman was investigated because of productive cough and 42 years old man was investigated because of haemoptysis. In two cases, at bronchoscopy, the typical picture of TO was observed. Microscopic examination of the biopsy material revealed bone formation. TO should be considered in the differential diagnosis as an unusual cause of chronic persistent cough, haemoptysis, persistent atelectasis, and recurrent segmental or lobar infection. PMID- 15351942 TI - Primary endobronchial leiomyoma: a case report. AB - Pulmonary leiomyoma is a rare benign tumor. It has been usually described as a single case. The aim of this paper is to present a case of endobronchial leiomyoma. A 43 year-old nonsmoker female presented to our center with complaint of cough and sputum production for two years. Her chest roentgenogram showed consolidation on lower zone of right lung. Computed tomography of thorax demonstrated a mass lesion partially obstructing lateral segment bronchus of right middle lobe and consolidation on right lower lung field. Fiberoptic bronchoscopy detected mass obstructing lateral segment bronchus of right middle lobe. The biopsy specimen obtained by fiberbronchoscopy revealed the diagnosis of endobronchial leiomyoma. There was no pathologic finding in uterine examination. Bilobectomy inferior was performed. The patient was healthy six months later. PMID- 15351943 TI - [Recurrent pulmonary embolism secondary to hereditary thrombophilia (case report)]. AB - Pulmonary embolism is seen commonly but diagnosed difficulty and has high mortality. There are too many risk factors that have been described for pulmonary embolism. However, the hereditary factors are important risk factors for the cases especially with recurrent pulmonary embolism. In our case who had been diagnosed as pulmonary embolism three times before and treated with anticoagulants, because of the recurrent pulmonary embolism, the genetic risk factors were investigated. Homozygous factor V Leiden mutation, deficiency of protein S and hyperhomocysteinaemia were determined in our case. In addition, in the investigation of the family, protein S, protein C and factor V Leiden mutation were determined in all three daughters of our case. Since our patient has recurrent pulmonary embolism and has more than one genetic risk factors, anticoagulant treatment was planned for lifelong. Recurrent thromboembolism is too important because of threatening the life. Identification of the genetic risk factors that result in increased tendency to thrombosis has important implications for the patients and their families. PMID- 15351944 TI - [Bronchiolitis obliterans organizing pneumonia presenting with solitary pulmonary nodule and spontan pneumothorax]. AB - Chronic obstructive pulmonary disease (COPD) is characterized with progressive airflow limitation as a result of abnormal inflammation due to inhalation of various noxious gases and particulate dusts. COPD is an increasing important health problem that is parallel to the increasing habit of tobacco smoke. Tracheobronchial infections seem to be the most important cause of exacerbation in COPD, however pneumothorax and pulmonary thromboembolism are also important determinant factors in the attack's severity. Since the tobacco smoking is the common risk factor in both COPD and lung cancer, solitary pulmonary nodules especially in smokers should be carefully examined. Bronchiolitis obliterans organizing pneumonia (BOOP) is an uncommon pulmonary disorder, the clinical spectrum of which is variable. In this case report, a COPD patient manifesting spontaneous pneumothorax and solitary pulmonary nodule has been discussed who was diagnosed as BOOP after surgical procedure. PMID- 15351945 TI - [Approach of diffuse interstitial lung diseases]. AB - Diffuse interstitial lung diseases (DILD) are heterogeneous set of diseases affecting the interstitium of the lung. The etiologic factors may be known or unknown. The diagnosis is based on the disease history, findings of the physical and radiological examinations, physiological and bronchoscopic findings, the cyto and/or histopathological findings of the bronchoscopic materials and surgical lung biopsy in some selected patients. The reader will find the definition, classification, general properties and clinical approach of DILD. PMID- 15351946 TI - Rights of the children against tobacco (protect them before they are misleaded, educate them before they take up the habit). PMID- 15351947 TI - [Idiopathic retroperitoneal fibrosis]. AB - Retroperitoneal fibrosis is an uncommon disease, characterized by the replacement of normal retroperitoneal tissue with fibrosis and/or chronic inflammation. In two thirds of the cases retroperitoneal fibrosis is idiopathic (IRF), whereas in the remaining ones it is secondary/associated to cancer, infections, drugs, autoimmune disease and vasculitis. IRF appears as a dense, fibrous plaque that usually arises between the level of the lower aorta and the common iliac arteries. As the plaque progresses, it engulfs the adjacent structures (e. g., ureters). In its early stages IRF is characterized by a rich infiltrate of lymphocytes, plasma cells and macrophages interspersed within fibroblasts and collagen bundles. In its advanced stages it becomes relatively avascular and acellular with abundant collagen bundles and scattered calcifications. The pathogenesis is unknown: some Authors suggest that IRF is a consequence of a local autoimmune reaction against atherosclerotic plaque antigens whereas others propose that it is the manifestation of a systemic autoimmune disease. The presenting signs and symptoms are non-specific; systemic manifestations (fever, anorexia, weight loss), often associated with local symptoms, are usually found to be related to the entrapment of retroperitoneal structures. The most common local symptom is lumbar and/or abdominal pain. The treatment can be surgical and/or medical: the former is required when obstructive complications are present; the latter, associated or not with surgery, can significantly improve the outcome of IRF patients and usually modifies the natural history of the disease. Steroids and tamoxifen are the most used drugs, whereas other agents such as azathioprine, methotrexate and cyclosporine are usually given to non responder patients. PMID- 15351948 TI - [Familial hyperaldosteronism]. AB - Primary aldosteronism is a disorder characterized by hypertension and hypokalemia due to aldosterone secretion out of renin-angiotensin control. It is generally caused by aldosterone-producing adenoma or adrenocortical hyperplasia but, in some cases, it is due to genetic alterations. Familial type I hyperaldosteronism is the result of anomalous regulation of aldosterone secretion from ACTH (which normally regulates cortisol synthesis). Aldosterone hypersecretion can be suppressed by exogenous glucocortcoids such as dexamethasone. This autosomal dominant disorder is caused by unequal cross-over between two genes with wide sequence homology: CYP11B1 and CYP11B2. The hybrid gene is the product of fusion between the ACTH-responsive regulatory portion of the 11b-hydroxylase gene (CYP11B1) and the coding region of the aldosterone synthase gene (CYP11B2). Familial type I hyperaldosteronism is a disease with incomplete penetration and variable expressivity, especially in relation to hypertension. The marked variability in hypertension severity can mirror an interaction between the hybrid gene and other hereditary factors involved in the regulation of blood pressure. Familial type II hyperaldosteronism is another autosomal dominant form of hyperaldosteronism due to aldosterone hyper-secretion not suppressible by dexamethasone. This disorder is unrelated to mutation of the hybrid gene. The genetic cause of type II hyperaldosteronism is presently unknown, but a genome wide search has revealed that the disorder is linked with a locus on chromosome 7 in a region that corresponds to cytogenetic band 7p22. PMID- 15351949 TI - [Cardiovascular risk and renal transplantation]. AB - Cardiovascular disease after renal transplantation is often the expression of a disease process that first started with the onset of renal dysfunction many years before, and its prevention starts with the early predialysis phase of chronic renal failure and with the aggressive treatment of hypertension and dyslipidemia. The evidence that dialysis treatment itself accelerates arterial damage is poor. After transplantation, however, many patients are restored to a state not of normal renal function but of chronic renal impairment and have drug-induced hypertension and dyslipidemia, resulting in a vastly increased risk of atherosclerosis. Further research is required on optimal strategies to prevent or ameliorate cardiovascular disease, to establish the roles of lipid-lowering and antihypertensive therapies after transplantation and to define immunosuppressive ad hoc treatments for each kind of patient. PMID- 15351950 TI - [Clinical dialysis: new problems and new prospects]. AB - The main problem nephrologists have to face today is the very high patient morbidity and mortality. A number of traditional and non-traditional risk factors have a role; among these anaemia, hypertension, dislipidemia, abnormalities in calcium-phosphate metabolism, hyperhomocysteinemia and endothelial dysfunction. An important innovation in the field of hemodialysis has been the availability of high-permeable and high-flux membranes, characterized by a high biocompatibility and ultrafiltration coefficient. The development of automatic systems to control ultrafiltration has enabled the utilisation of these membranes in the clinical setting (high-flux hemodialysis, hemofiltration, hemodiafiltration). It is common opinion that high-flux membranes can positively influence cardiovascular instability, but this has not been confirmed by clinical trials. Although preliminary data indicated a favorable effect on the correction of anemia in patients treated with high-permeable membranes, randomized trials have not shown a significant effect. Better control of anemia could be possible by means of on line treatments, given their higher removal of medium- and large molecules and reduced microbiological and pyrogenic contamination of the dialysate. A number of analyses showed a lower incidence of bone cysts and/or carpal tunnel syndrome in patients treated with high-flux membranes compared to low-flux ones. High-flux treatments could reduce morbidity and mortality in hemodialysis patients. However, despite its large sample size, the HEMO Study has not been capable of showing a statistically significant effect of higher dialysis dose and high-flux membranes on survival and morbidity. The MPO study has been expressively designed to do a prospective evaluation of the long-term effect of membrane permeability on clinical outcomes. These results are greatly awaited. PMID- 15351951 TI - [Microhematuria. Prevalence, test validity and clinical usefulness]. AB - The dipstick test is the most sensitive test for microhematuria screening. In the general population asymptomatic microhematuria is not rare. In terms of clinical presentation hematuria can be classified in a double temporal dimension, i.e. according to the age of the patients and to its time course (resolution vs. persistence). In isolated microhematuria, red cells morphology is useful to establish the site of hematuria (glomerular vs. non glomerular). This is important mostly because in older patients this alteration may underlie a genito urinary cancer. Microhematuria is probably one of the most sought for clinical signs of urinary diseases. It is fundamental that this clinical sign be always interpreted in a context considering other symptoms and clinical data. PMID- 15351952 TI - [Bilateral hydronephrosis: a stringent diagnosis]. AB - A forty-nine year old man was admitted to the Renal Unit of our Hospital because of renal failure, loin pain, low grade fever and loss of weight. It was done a complete work-up. The ultrasound and the computed tomography of the abdomen were very helpful to get the right diagnosis. PMID- 15351954 TI - Stratification and weighting via the propensity score in estimation of causal treatment effects: a comparative study. AB - Estimation of treatment effects with causal interpretation from observational data is complicated because exposure to treatment may be confounded with subject characteristics. The propensity score, the probability of treatment exposure conditional on covariates, is the basis for two approaches to adjusting for confounding: methods based on stratification of observations by quantiles of estimated propensity scores and methods based on weighting observations by the inverse of estimated propensity scores. We review popular versions of these approaches and related methods offering improved precision, describe theoretical properties and highlight their implications for practice, and present extensive comparisons of performance that provide guidance for practical use. PMID- 15351955 TI - Estimating vaccine efficacy from household data observed over time. AB - Estimation of vaccine efficacy has traditionally focused on the reduction in susceptibility to infection, or the vaccine efficacy for susceptibility (VE(S)). However, a vaccine, such as a prophylactic HIV vaccine, may also lower the infectiousness of a vaccinated person who became infected. The relative reduction in infectiousness due to vaccination is the vaccine efficacy for infectiousness (VE(I)). Estimation of VE(I) is challenging because it requires information on exposure to infection, and gathering this type of information is often expensive and difficult, or even impossible. Household studies are expected to provide more information on who is exposed to whom. In a previous paper, we developed a method for estimating VE(S) and VE(I) from a household study where only the final outbreak data are available. However, the resulting estimates were quite unstable. In this work, we develop a survival model for the estimation of VE(S) and VE(I) from household data where the time of infection is known for every study participant. Using stochastic simulations, we show that the proposed method significantly reduces the bias and mean square error in the estimation of both VE(S) and VE(I) as compared to the method based on final outbreak data. We also show that when time-to-event data are available, a household study produces more robust estimators than a same-size study of unrelated individuals. In addition, we investigate the bias in estimating VE(S) and VE(I) due to misclassification of infection status when only illness data, rather than true infection data, are available. PMID- 15351956 TI - Multiple comparisons with a control in families with both one-sided and two-sided hypotheses. AB - Comparing several treatments with a control is a common objective of clinical studies. However, existing procedures mainly deal with particular families of inferences in which all hypotheses are either one- or two-sided. In this article, we seek to develop a procedure which copes with a more general testing environment in which the family of inferences is composed of a mixture of one- and two-sided hypotheses. The proposed procedure provides a more flexible and powerful tool than the existing method. The superiority of this method is also substantiated by a simulation study of average power. Selected critical values are tabulated for the implementation of the proposed procedure. Finally, we provide an illustrative example with sample data extracted from a medical experiment. PMID- 15351957 TI - Improving Cox survival analysis with a neural-Bayesian approach. AB - In this article we show that traditional Cox survival analysis can be improved upon when supplemented with sensible priors and analysed within a neural Bayesian framework. We demonstrate that the Bayesian method gives more reliable predictions, in particular for relatively small data sets. The obtained posterior (the probability distribution of network parameters given the data) which in itself is intractable, can be made accessible by several approximations. We review approximations by Hybrid Markov Chain Monte Carlo sampling, a variational method and the Laplace approximation. We argue that although each Bayesian approach circumvents the shortcomings of the original Cox analysis, and therefore yields better predictive results, in practice the use of variational methods or Laplace is preferable. Since Cox survival analysis is infamous for its poor results with (too) many inputs, we use the Bayesian posterior to estimate p values on the inputs and to formulate an algorithm for backward elimination. We show that after removal of irrelevant inputs Bayesian methods still achieve significantly better results than classical Cox. PMID- 15351958 TI - Modelling infectious disease transmission with complex exposure pattern and sparse outcome data. AB - We present a regression modelling framework to analyse infectious disease transmission during a time period where extensive exposure data are available, but where the outcome data are sparse. A latent variable model is used for each exposure time, allowing a straight-forward accumulation of risk for a collection of exposures for which outcome data are available. We describe an analysis of HIV infection from blood products among a cohort of haemophiliacs in Ireland between 1980 and 1985. The analysis provides estimates of the time pattern and batch effects; we show how analytical complexity such as smoothly varying coefficients or random coefficient models can be accommodated by the model. Finally, we discuss other problems where the model is applicable. PMID- 15351959 TI - A statistical model for high-resolution mapping of quantitative trait loci determining HIV dynamics. AB - Are there specific genes that control the pathogenesis of HIV infection? This question, which is of fundamental importance in designing personalized strategies of gene therapy to control HIV infection, can be examined by genetic mapping approaches. In this article, we present a new statistical model for unravelling the genetic mechanisms for the dynamic change of HIV that causes AIDS by marker based linkage disequilibrium (LD) analyses. This new model is the extension of our functional mapping theory to integrate viral load trajectories within a genetic mapping framework. Earlier studies of HIV dynamics have led to various mathematical functions for modelling the kinetic curves of plasma virions and CD4 lymphocytes in HIV patients. Through incorporating these functions into the LD based mapping procedure, we can identify and map individual quantitative trait loci (or QTL) responsible for viral pathogenesis. We derive a closed-form solution for estimating QTL allele frequency and marker-QTL linkage disequilibrium in the context of EM algorithm and implement the simplex algorithm to estimate the mathematical parameters describing the curve shapes of HIV pathogenesis. We performed different simulation scenarios based on currently used clinical designs in AIDS/HIV research to illustrate the utility and power of our model for genetic mapping of HIV dynamics. The implications of our model for genetic and genomic research into AIDS pathogenesis are discussed. PMID- 15351960 TI - Smooth centile curves for skew and kurtotic data modelled using the Box-Cox power exponential distribution. AB - The Box-Cox power exponential (BCPE) distribution, developed in this paper, provides a model for a dependent variable Y exhibiting both skewness and kurtosis (leptokurtosis or platykurtosis). The distribution is defined by a power transformation Y(nu) having a shifted and scaled (truncated) standard power exponential distribution with parameter tau. The distribution has four parameters and is denoted BCPE (mu,sigma,nu,tau). The parameters, mu, sigma, nu and tau, may be interpreted as relating to location (median), scale (approximate coefficient of variation), skewness (transformation to symmetry) and kurtosis (power exponential parameter), respectively. Smooth centile curves are obtained by modelling each of the four parameters of the distribution as a smooth non parametric function of an explanatory variable. A Fisher scoring algorithm is used to fit the non-parametric model by maximizing a penalized likelihood. The first and expected second and cross derivatives of the likelihood, with respect to mu, sigma, nu and tau, required for the algorithm, are provided. The centiles of the BCPE distribution are easy to calculate, so it is highly suited to centile estimation. This application of the BCPE distribution to smooth centile estimation provides a generalization of the LMS method of the centile estimation to data exhibiting kurtosis (as well as skewness) different from that of a normal distribution and is named here the LMSP method of centile estimation. The LMSP method of centile estimation is applied to modelling the body mass index of Dutch males against age. PMID- 15351961 TI - Multiple imputation for body mass index: lessons from the Australian Longitudinal Study on Women's Health. AB - In large epidemiological studies missing data can be a problem, especially if information is sought on a sensitive topic or when a composite measure is calculated from several variables each affected by missing values. Multiple imputation is the method of choice for 'filling in' missing data based on associations among variables. Using an example about body mass index from the Australian Longitudinal Study on Women's Health, we identify a subset of variables that are particularly useful for imputing values for the target variables. Then we illustrate two uses of multiple imputation. The first is to examine and correct for bias when data are not missing completely at random. The second is to impute missing values for an important covariate; in this case omission from the imputation process of variables to be used in the analysis may introduce bias. We conclude with several recommendations for handling issues of missing data. PMID- 15351962 TI - Control of asymmetric cell divisions: will cnidarians provide an answer? AB - Cells in the basal metazoan phylum Cnidaria are characterized by remarkable plasticity in their differentiation capacity. The mechanism controlling asymmetric cell divisions is not understood in cnidarians or in any other animal group. PIWI proteins recently have been shown to be involved in maintaining the self-renewal capacity of stem cells in organisms as diverse as ciliates, flies, worms and mammals. Seipel et al.1 find that, in the cnidarian Podocoryne carnea, the Piwi homolog Cniwi is transcriptionally upregulated when the polyp generates buds, which will develop into medusae. Since transdifferentiation of striated muscle cells to smooth muscle cells also activated Cniwi expression, Cniwi appears to play a crucial role in differentiation events. The discovery should facilitate elucidation of the poorly understood factors that control asymmetric cell divisions at the beginning of animal evolution. PMID- 15351963 TI - Bile ducts as a source of pancreatic beta cells. AB - In recent years, there have been a number of well-documented examples demonstrating that one cell type can be converted to another. Two such examples are the appearance of ectopic pancreas in the liver and formation of hepatic tissue in the pancreas. The conversion of liver to pancreas raises the intriguing possibility of generating insulin-producing beta cells for therapeutic transplantation into diabetics. There is now a striking addition to the growing list of pancreatic conversions: the formation of pancreatic tissue in the developing biliary system. PMID- 15351964 TI - Class III HD-Zip gene regulation, the golden fleece of ARGONAUTE activity? AB - MicroRNAs, small noncoding RNAs, are implicated in gene regulation in both metazoans and plants. In plants, many of the targets of miRNA-mediated gene regulation encode transcription factors with functions in development, such as the Class III HD-Zip gene family whose members direct polarity establishment in leaves and vasculature. Three recent papers provide insight into how miRNAs, likely acting through a complex containing an Argonaute protein, regulate Class III HD-Zip gene expression in Arabidopsis and maize.1-3 While the precise biological activity of Argonaute proteins remains an enigma, ARGONAUTE1 in Arabidopsis is required for the proper regulation of miRNA165/166, which targets cleavage of Class III HD-Zip mRNAs. Consistent with their proposed role in negative regulation, expression of miRNA165/166 is complementary to that of Class III HD-Zip gene expression, but this is perturbed in agronaute1 mutants. Determining how these complementary patterns of expression are formed should lead us closer to an understanding of the molecular mechanisms by which asymmetry is established in developing leaves. PMID- 15351965 TI - VEGF: once regarded as a specific angiogenic factor, now implicated in neuroprotection. AB - Both blood vessels and nerves are guided to their target. Vascular endothelial growth factor (VEGF)A is a key signal in the induction of vessel growth (a process termed angiogenesis). Though initial studies, now a decade ago, indicated that VEGF is an endothelial cell-specific factor, more recent findings revealed that VEGF also has direct effects on neural cells. Genetic studies showed that mice with reduced VEGF levels develop adult-onset motor neuron degeneration, reminiscent of the human neurodegenerative disorder amyotrophic lateral sclerosis (ALS). Additional genetic studies confirmed that VEGF is a modifier of motor neuron degeneration in humans and in SOD1(G93A) mice--a model of ALS. Reduced VEGF levels may promote motor neuron degeneration by limiting neural tissue perfusion and VEGF-dependent neuroprotection. VEGF also affects neuron death after acute spinal cord or cerebral ischemia, and has also been implicated in other neurological disorders such as diabetic and ischemic neuropathy, nerve regeneration, Parkinson's disease, Alzheimer's disease and multiple sclerosis. These findings have raised growing interest in assessing the therapeutic potential of VEGF for neurodegenerative disorders. PMID- 15351966 TI - Adding to the ends: what makes telomerase processive and how important is it? AB - Telomerase is a cellular reverse transcriptase responsible for telomere maintenance in most organisms. It does so by adding telomere repeats onto pre existing ends using an integral RNA component as template. Compared to "prototypical" reverse transcriptases, telomerase is unique in being able to repetitively copy a short templating RNA segment, thus adding multiple copies of the repeat to the DNA substrate following a single binding event. This uniquely processive property hints at the intricate conformational alterations that the enzyme must choreograph during its reaction cycles. Recent studies have identified distinct structural elements within both the RNA and protein components of telomerase that modulate enzyme processivity. Pharmacological and genetic analysis suggest that telomerase processivity is a significant determinant of telomere length. Because telomere maintenance and the lack thereof have been linked to tumor progression and aging, further investigation of telomerase processivity may lead to novel medical intervention strategies. PMID- 15351967 TI - PML nuclear bodies: dynamic sensors of DNA damage and cellular stress. AB - Promyelocytic leukaemia nuclear bodies (PML NBs) are generally present in all mammalian cells, and their integrity correlates with normal differentiation of promyelocytes. Mice that lack PML NBs have impaired immune function, exhibit chromosome instability and are sensitive to carcinogens. Although their direct role in nuclear activity is unclear, PML NBs are implicated in the regulation of transcription, apoptosis, tumour suppression and the anti-viral response. An emerging view is that they represent sites where multi-subunit complexes form and where post-translational modification of regulatory factors, such as p53, occurs in response to cellular stress. Following DNA damage, several repair factors transit through PML NBs in a temporally regulated manner implicating these bodies in DNA repair. We propose that PML NBs are dynamic sensors of cellular stress, which rapidly disassemble following DNA damage into large supramolecular complexes, dispersing associated repair factors to sites of damage. The dramatically increased total surface area available would enhance interactions between PML-associated factors regulating DNA repair and apoptosis. PMID- 15351968 TI - Dynamic cross-talk between cells and the extracellular matrix in the testis. AB - In the seminiferous tubule of the mammalian testis, one type A1 spermatogonium (diploid, 2n) divides and differentiates into 256 spermatozoa (haploid, n) during spermatogenesis. To complete spermatogenesis and produce approximately 150 x 10(6) spermatozoa each day in a healthy man, germ cells must migrate progressively across the seminiferous epithelium yet remain attach to the nourishing Sertoli cells. This active cell migration process involves precisely controlled restructuring events at the tight (TJ) and anchoring junctions at the cell-cell interface. While the hormonal events that regulate spermatogenesis by follicle-stimulating hormone and testosterone from the pituitary gland and Leydig cells, respectively, are known, less is known about the mechanism(s) that regulates junction restructuring during germ cell movement in the seminiferous epithelium. The relative position of tight (TJs) and anchoring junctions in the testis is of interest. Sertoli cell TJs that constitute the blood-testis barrier (BTB) are present side by side with anchoring junctions and are adjacent to the basement membrane. This intimate physical association with the TJs, the anchoring junctions and the basement membrane (a modified form of extracellular matrix, ECM) suggests a role for the ECM in the junction dynamics of the testis. Indeed, evidence is accumulating that ECM proteins are crucial to Sertoli cell TJ dynamics. In this review, we discuss the pivotal role of tumor necrosis factor alpha (TNFalpha) on BTB dynamics via its effects on the homeostasis of ECM proteins. In addition, discussion will also be focused on the novel findings regarding the role of non-basement-membrane-associated ECM proteins and components of focal adhesion (a cell-matrix anchoring junction type) in the regulation of junction dynamics in the testis. PMID- 15351969 TI - Trafficking and signaling pathways of nuclear localizing protein ligands and their receptors. AB - Interaction of ligands such as epidermal growth factor and interferon-gamma with the extracellular domains of their plasma membrane receptors results in internalization followed by translocation into the nucleus of the ligand and/or receptor. There has been reluctance, however, to ascribe signaling importance to this, the focus instead being on second messenger pathways, including mobilization of kinases and inducible transcription factors (TFs). The latter, however, fails to explain the fact that so many ligands stimulate the same second messenger cascades/TFs, and yet show distinct gene activation profiles. This is particularly apt in the case of the seven STAT TFs that are held to be the mediators of the distinct cellular functions of over 60 ligands. The current review focuses on five representative nuclear localizing ligands for which there is documentation of translocation into the cytosol and nucleus through well characterized pathways, in addition to a role in gene activation by ligand/receptor in the nucleus. PMID- 15351970 TI - Model systems in stem cell biology. AB - Stem cell scientists and ethicists have focused intently on questions relevant to the developmental stage and developmental capacities of stem cells. Comparably less attention has been paid to an equally important set of questions about the nature of stem cells, their common characteristics, their non-negligible differences and their possible developmental species specificity. Answers to these questions are essential to the project of justly inferring anything about human stem cell biology from studies in non-human model systems--and so to the possibility of eventually developing human therapies based on stem cell biology. After introducing and discussing these questions, I conclude with a brief discussion of the creation of novel model systems in stem cell biology: human-to animal embryonic chimeras. Such novel model systems may help to overcome obstacles to extrapolation, but they are also scientifically and ethically contentious. PMID- 15351971 TI - Stem cells as probabilistic self-producing entities. AB - Stem cells have the capacity both to self-renew and to give rise to differentiated progeny, and are vital to the organization of multicellular organisms. Stem cells raise a number of fundamental questions regarding lineage restriction and cellular differentiation, and they hold enormous promise for cell based therapies. Here I propose a theoretical framework for stem cell biology based on the concepts of autopoiesis (self-production) and complementarity. I argue that stem cells are pivotal in the self-production of the organism and that we need complementary approaches to understand their probabilistic behavior. I discuss how this framework generates testable hypotheses regarding stem-cell functions. PMID- 15351972 TI - Molecular mechanisms for organizing the neuronal cytoskeleton. AB - Neurofilaments and microtubules are important components of the neuronal cytoskeleton. In axons or dendrites, these filaments are aligned in parallel arrays, and separated from one another by nonrandom distances. This distinctive organization has been attributed to cross bridges formed by NF side arms or microtubule-associated proteins. We recently proposed a polymer-brush-based mechanism for regulating interactions between neurofilaments and between microtubules. In this model, the side arms of neurofilaments and the projection domains of microtubule-associated proteins are highly unstructured and exert long range repulsive forces that are largely entropic in origin; these forces then act to organize the cytoskeleton in axons and dendrites. Here, we review the biochemical, biophysical, genetic and cell biological data for the polymer-brush and cross-bridging models. We explore how the data traditionally used to support cross bridging may be reconciled with a polymer-brush mechanism and compare the implications of recent experimental insights into axonal transport and physiology for each model. PMID- 15351973 TI - The gain in the brain is plain when evo meets devo. PMID- 15351974 TI - Gender transitions. PMID- 15351975 TI - African genomics. PMID- 15351976 TI - Coexpression of p53 protein and MDR functional phenotype in leukemias: the predominant association in chronic myeloid leukemia. AB - BACKGROUND: One of the best characterized resistance mechanisms of leukemias is multidrug resistance (MDR) mediated by P-glycoprotein (Pgp) and multidrug resistant related protein (MRP). In addition to Pgp and MRP, p53 mutation or inactivation might play a relevant role in therapeutic failure. Some studies have demonstrated that Pgp and MRP may be activated in association with overexpression of mutant or inactivated p53 protein. The aim of this study was to investigate the association between p53 expression and MDR functional phenotype analyzed by flow cytometry (FCM). METHODS: Rhodamine-123 assay analyzed by FCM was used to detect the MDR phenotype that was positive in 18 out of 41 (43.9%) cases of chronic myeloid leukemia (CML), 16 out of 28 (57.1%) chronic lymphoid leukemia (CLL) cases, 11 out of 28 (39.3%) acute myeloid leukemia (AML) cases, and four out of 22 (18.2%) acute lymphoid leukemia (ALL) cases. RESULTS: Variable levels of p53 expression were observed in leukemic cells: 12 out of 41 (29.2%) in CML, nine out of 28 (32.1%) in CLL, 15 out of 28 (53.6%) in AML, and eight out of 22 (36.4%) in ALL samples. CONCLUSIONS: In our study, no significant association between p53 expression and MDR functional phenotype was observed in ALL, CLL, and AML. On the other hand, a significant association (P = 0.0003) of the coexpression was observed in CML. The p53 overexpression was more frequently seen in the accelerated phase and the blastic phase of this disease. Our results suggest that an MDR functional phenotype could be associated with p53 mutation in the advanced stage of leukemias. PMID- 15351977 TI - Detection of residual neuroblastoma cells in bone marrow: comparison of flow cytometry with immunocytochemistry. AB - BACKGROUND: Because the cytomorphologic examination of bone marrow (BM) aspirates appears not sensitive enough to detect residual neuroblastoma cells, two four color flow cytometric assays using different combinations of CD9, CD81, CD56, CD45, and anti-GD2 were evaluated. METHODS: The sensitivity of the flow cytometric assays was assessed by spiking experiments in normal peripheral blood samples. Twenty-eight BM samples, 12 biopsies, and 3 peripheral blood stem cell (PBSC) preparations from 22 patients with neuroblastoma were analyzed. The results were compared with those of an anti-GD2 immunocytochemical reference assay. RESULTS: Flow cytometric and immunocytochemical analyses showed residual neuroblastoma cells in four BM samples. One PBSC preparation and 20 BM samples were negative for both assays. Four BM and two PBSC samples scored positive for the immunocytochemical assay but were negative for the flow cytometric tests. This was due to the limited number of cells that were flow cytometrically analyzed. A strong correlation between the flow cytometric and immunocytochemical tests was found (chi2 = 6.4, P = 0.011). CONCLUSIONS: When an equal amount of cells is analyzed, the sensitivity of the flow cytometric assays is to be about 10 times lower than that of the immunocytochemical test. However, the flow cytometric assays can be used to screen for residual cells in clinical samples with a sensitivity of one neuroblastoma cell in 10(4) to 10(5) normal mononuclear cells. Flow cytometry is simple, quick, and cost effective compared with immunocytochemistry. In addition, the flow cytometric assays can be used to screen for residual neuroblastoma cells in case of a GD2-negative primary tumor. Therefore we recommend flow cytometry for the detection of residual neuroblastoma cells. PMID- 15351978 TI - Comparison of CD4 and CD8 lymphocyte counts in HIV-negative pulmonary TB patients with those in normal blood donors and the effect of antitubercular treatment: hospital-based flow cytometric study. AB - BACKGROUND: This study was designed to flow cytometrically determine baseline and sequential values of CD4 and CD8 lymphocyte subsets in patients without the human immunodeficiency virus and with pulmonary tuberculosis (TB) and to correlate these values with those obtained from normal male blood donors and with the radiologic extent of disease and response to therapy. METHODS: We studied 39 male patients without the human immunodeficiency virus and with sputum positive for pulmonary TB who had been admitted to Military Hospital (Cardiothoracic Center) in Pune, India. Clinical, laboratory, and radiologic evaluations of these patients were done. Hematologic parameters were assessed by an automated hematology cell counter (AcT*Diff, Coulter), and T-cell subsets (CD4 and CD8) were determined flow cytometrically (EPICS-XL, Coulter). RESULTS: CD4 counts and percentages of CD4 were significantly lower, but CD8 values were normal, in patients with pulmonary TB when compared with values obtained in normal blood donors. The CD4/CD8 ratio was significantly lower in patients with TB. The CD4 counts normalized with antitubercular treatment. The radiologic extent of disease did not correlate well with the immune parameters studied. CONCLUSIONS: TB is a reversible cause of CD4 lymphocytopenia and is associated with normal numbers of CD8 cells. The radiologic extent of disease does not seem to determine the immune response. PMID- 15351979 TI - Flow cytometric characterization of cell populations in bronchoalveolar lavage and bronchial brushings from patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a serious, chronic inflammatory disease of the airway associated with cigarette smoking. Leucocytes are involved in the inflammatory process in the airways in COPD. There is a need for accurate characterization of cellular populations in bronchoalveolar lavage (BAL) due to variation in the predominant cell types reported, which were investigated mostly with manual counting techniques. METHODS: Bronchial brushings and BAL were obtained from human subjects undergoing fiber optic bronchoscopy. Flow cytometry was applied to identify various cell types. Quenching of autofluorescence of BAL-derived alveolar macrophages was achieved with eta-octyl beta-D-galactopyranoside and crystal violet. Comparisons of cell counts obtained with flow cytometric and manual counting methods were performed. RESULTS: Correlation analysis showed that manual cell counting methods overestimated the percentage of macrophages when compared with flow cytometric methods (R2 = 0.54). There was also a small tendency by manual counting to underestimate the percentage of lymphocytes and neutrophils. Using flow cytometry, the percentage and absolute numbers of alveolar macrophages and lymphocytes in BAL were not significantly different between patients with COPD and control subjects. The percentage and absolute numbers of neutrophils were higher in BAL from patients with moderate to severe COPD. CONCLUSIONS: This novel flow cytometric assay for identification of various cell types from heterogenous samples of BAL and bronchial brushing will allow further investigation of cell characteristics, such as cytokine production and receptor expression, and an accurate evaluation of apoptosis for different cell types and provide a rationale for urgently required effective treatments for COPD. PMID- 15351980 TI - Multiplex bead array assays for detection of soluble cytokines: comparisons of sensitivity and quantitative values among kits from multiple manufacturers. AB - BACKGROUND: Multiplex bead array assays permit simultaneous cytometric quantitation of multiple cytokines in solution by capturing these to spectrally distinct beads. Because several manufacturers offer reagents to quantitate the same cytokines on a single instrument, a comparison should be made to determine whether these kits yield similar data and whether these data are comparable to enzyme-linked immunosorbent assay (ELISA). METHODS: This study compared cytokine detection kits by using Luminex 100. Twenty-six serum samples from seven subjects were analyzed for interferon-gamma, interleukins 1beta, 6, and 8, and tumor necrosis factor-alpha by using multiplex kits from LINCO Research, Bio-Rad Laboratories, R&D Systems, and BioSource International. Each assay was performed according to the manufacturers' specifications. Standard curves were generated by using reference concentrations supplied by each manufacturer. ELISAs for interleukin-8 were performed by using kits from R&D and BioSource. RESULTS: Cytokine levels followed similar patterns, although absolute concentrations differed among kits. ELISA and Luminex values for interleukin-8 were similar in kits from the same manufacturer. CONCLUSIONS: Because relative cytokine measurements are often valuable when performed serially, it may be possible to make interlaboratory comparisons by using different kits. When comparison of absolute values is crucial, kits from the same supplier should be used. Within vendor, bead array, and ELISA values appear comparable. PMID- 15351981 TI - Flow cytometry detection of Shiga toxins in the blood from children with hemolytic uremic syndrome. AB - BACKGROUND: Hemolytic uremic syndrome (HUS) is the main cause of acute renal failure in early childhood. Most cases are due to intestinal infections from Escherichia coli strains (STEC) which produce by Shiga toxin (Stxs). Stx1 and Stx2 produced by STEC in the gut are absorbed into the circulation and, after binding on polymorphonuclear leukocytes (PMNs), are targeted to renal endothelium. The aim of the present work was the development of a method to detect Stxs bound on circulating PMNs and to diagnose STEC infections in patients with HUS. METHODS: White blood cells isolated after erythrocytic lysis were incubated with anti-Stxs mouse monoclonal antibodies in the presence of human serum to saturate Fc receptors on PMNs. After incubation with fluorescein isothiocyanate-goat anti-mouse immunoglobulin G, flow cytometric analysis was used to demonstrate the cell-bound fluorescence. RESULTS: The method was quick (3 h), sensitive (femtomoles), and capable of detecting both Stxs. The presence of Stxs was detected on PMNs from six patients with HUS: four patients had serologic or microbiological evidence of STEC infection, whereas the other two patients had no evidence of STEC infection when employing the standard diagnostic methods. CONCLUSIONS: The method described is rapid, simple, and based on commercially available reagents, and it might be more sensitive than the standard methods for diagnosis of STEC infection. It also allows the detection of Stxs in blood, a key step to monitor the pathogenesis of HUS. PMID- 15351982 TI - CD3 bright lymphocyte population reveal gammadelta T cells. AB - BACKGROUND: In routine CD3/CD4/CD8 T-cell analysis, a CD3 bright population of lymphocytes is frequently observed. The aim of the present study was to identify the immunological significance of such CD3 bright lymphocytes. METHODS: We analyzed samples from 31 healthy adult volunteers, 78 human immunodeficiency virus (HIV)-positive, and 78 renal transplanted patients. RESULTS: A clearly distinct CD3 bright (frequently CD4-/CD8-) T-cell fraction was observed in 84% of donors and was directly correlated with the fraction of gammadelta T cells (r2 = 0.64). CD3 overexpression on gammadelta T cells was confirmed by a combination of monoclonal antibody staining (CD3-ECD, gammadeltaTCR-FITC, and alphabetaTCR-PE Cy5) or immunomagnetic purification of gammadelta T cells (i.e., MdFI 20 vs 8.86). The gammadelta T cells expressed CD8 polypeptide chains (alpha and beta) in all possible combinations. The largest proportion, surprisingly, were cells expressing CD8betabeta homodimers (43.8 +/- 16.5%). CD8alphaalpha homodimers were expressed on 14.2% (+/- 12.3) of total gammadelta T cells, whereas CD8alphabeta heterodimers were expressed on 12.2% (+/- 7.5). We also observed a bimodal distribution of the intensity of CD3 fluorescence of gammadelta T cells in immunocompromised patients with a threshold at 105 cell/microl. CD3 bright gammadelta T cells were more frequently observed in HIV patients (29%) compared with renal transplant patients (11%) and healthy donors (3%; chi2 test: P = 0.0007). CONCLUSIONS: The simple observation of a CD3 bright T-cell subset on CD3/CD4/CD8 routine analysis suggests a high gammadelta T-cell fraction and, in our opinion, should be followed by a complementary analysis to determine precisely the number of gammadelta T cells and to identify their CD8alpha/beta phenotype. When CD3 bright T cells/microl were more than 40%, high gammadelta T cells were detected in more than 87% of cases, with a specificity of 76%. Occasionally, the CD3 bright subset appeared to be strongly homogeneous, suggesting an oligoclonal proliferation that could possibly reveal a chronic localized stimulation or an early lymphoproliferative disorder. Because the gammadelta T cells have interesting immunological peculiarities, the clinical significance of their quantitative abnormality should be clarified in diseases such as HIV, organ transplantation, autoimmunity and lymphoma. PMID- 15351983 TI - Scanning fluorescent microscopy analysis is applicable for absolute and relative cell frequency determinations. AB - BACKGROUND: Flow cytometry (FCM) and laser scanning cytometry (LSC) are the routine techniques for fluorescent cell analysis. Recently, we developed a scanning fluorescent microscopy (SFM) technique. This study compares SFM to LSC (two slide-based cytometry, SBC, techniques) and FCM, in experimental and clinical settings. METHODS: For the relative cell-frequency determinations, HT29 colorectal cancer cells and Ficoll separated blood mononuclear cells (FSBMCs) were serially diluted (from 1:1 to 1:1,000) and measured by each of the three techniques. For the absolute cell number determinations (only for SBC) FSBMCs were smeared on slides, then HT29 cells were placed on the slide with a micromanipulator (5-50 cells). Tumor cells circulating in the peripheral blood were isolated by magnetic separation from clinical blood samples of colorectal cancer patients. All samples were double-stained by CD45 ECD and CAM5.2 FITC antibodies. For slides, TOTO-3 and Hoechst 33258 DNA dyes were applied as nuclear counter staining. RESULTS: In the relative cell frequency determinations, the correlations between the calculated value and measured values by SFM, LSC, and FCM were r(2) = 0.79, 0.62, and 0.84, respectively (for all P < 0.01). In the absolute cell frequency determinations, SFM and LSC correlated to a high degree (r(2) = 0.97; P < 0.01). CONCLUSIONS: SFM proved to be a reliable alternative method, providing results comparable to LSC and FCM. SBC proved to be more suitable for rare-cell detection than FCM. SFM with digital slides may prove an acceptable adaptation of conventional fluorescent microscopes in order to perform rare-cell detection. PMID- 15351984 TI - Analysis of UV-excited fluorochromes by flow cytometry using near-ultraviolet laser diodes. AB - INTRODUCTION: Violet laser diodes have become common and reliable laser sources for benchtop flow cytometers. While these lasers are very useful for a variety of violet and some ultraviolet-excited fluorochromes (e.g., DAPI), they do not efficiently excite most UV-stimulated probes. In this study, the next generation of InGaN near-UV laser diodes (NUVLDs) emitting in the 370-375-nm range have been evaluated as laser sources for cuvette-based flow cytometers. METHODS: Several NUVLDs, ranging in wavelength from 370 to 374 nm and in power level from 1.5 to 10 mW, were mounted on a BD Biosciences LSR II and evaluated for their ability to excite cells labeled with the UV DNA binding dye DAPI, several UV phenotyping fluorochromes (including Alexa Fluor 350, Marina Blue, and quantum dots), and the fluorescent calcium chelator indo-1. RESULTS: NUVLDs at the 8-10-mW power range gave detection sensitivity levels comparable to more powerful solid-state and ion laser sources, using low-fluorescence microsphere beads as measurement standards. NUVLDs at all tested power levels allowed extremely high-resolution DAPI cell cycle analysis, and sources in the 8-10-mW power range excited Alexa Fluor 350, Marina Blue, and a variety of quantum dots at virtually the same signal-to-noise ratios as more powerful UV sources. CONCLUSIONS: These evaluations indicate that near-UV laser diodes installed on a cuvette-based flow cytometer performed nearly as well as more powerful solid-state UV lasers on the same instrumentation, and comparably to more powerful ion lasers on a jet-in-air system, and. Despite their limited power, integration of these small and inexpensive lasers into benchtop flow cytometers should allow the use of flow cytometric applications requiring UV excitation on a wide variety of instruments. PMID- 15351985 TI - The polarization of fluorescence of DNA stains depends on the incorporation density of the dye molecules. AB - BACKGROUND: The fluorescence induced by polarized light sources, such as the lasers that are used in flow cytometry, is often polarized and anisotropic. In addition, most optical detector systems are sensitive to the direction of polarization. These two factors influence the accuracy of fluorescence intensity measurements. The intensity of two light sources can be compared only if all details of the direction and degree of polarization are known. In a previous study, we observed that fluorescence polarization might be modified by dye-dye interactions. This report further investigates the role of dye density in fluorescence polarization anisotropy. METHODS: We measured the polarization distribution of samples stained with commonly used DNA dyes. To determine the role of fluorophore proximity, we compared the monomeric and a dimeric form of the DNA dyes ethidium bromide (EB), thiazole orange (TO), and oxazole yellow (YO). RESULTS: In all dyes sampled, fluorescence polarization is less at high dye concentrations than at low concentrations. The monomeric dyes exhibit a higher degree of polarization than the dimeric dyes of the same species. CONCLUSIONS: The polarization of fluorescence from DNA dyes is related to the density of incorporation into the DNA helix. Energy transfer between molecules that are in close proximity loosens the linkage between the excitation and emission dipoles, thereby reducing the degree of polarization of the emission. PMID- 15351986 TI - Whole genome amplification for CGH analysis: Linker-adapter PCR as the method of choice for difficult and limited samples. AB - BACKGROUND: Comparative genomic hybridization (CGH) is a powerful method to investigate chromosomal imbalances in tumor cells. However, DNA quantity and quality can be limiting factors for successful CGH analysis. The aim of this study was to investigate the applicability of degenerate oligonucleotide-primed PCR (DOP-PCR) and a recently developed linker-adapter-mediated PCR (LA-PCR) for whole genome amplification for use in CGH, especially for difficult source material. METHODS: We comparatively analyzed DNA of variable quality derived from different cell/tissue types. Additionally, dilution experiments down to the DNA content of a single cell were performed. FISH and/or classical cytogenetic analyses were used as controls. RESULTS: In the case of high quality DNA samples, both methods were equally suitable for CGH. When analyzing very small amounts of these DNA samples (equivalent to one or a few human diploid cells), DOP-PCR-CGH, but not LA-PCR-CGH, frequently produced false-positive signals (e.g., gains in 1p and 16p, and losses in chromosome 4q). In case of formalin-fixed paraffin embedded tissues, success rates by LA-PCR-CGH were significantly higher as compared to DOP-PCR-CGH. DNA of minor quality frequently could be analyzed correctly by LA-PCR-CGH, but was prone to give false-positive and/or false negative results by DOP-PCR-CGH. CONCLUSIONS: LA-PCR is superior to DOP-PCR for amplification of DNA for CGH analysis, especially in the case of very limited or partly degraded source material. PMID- 15351987 TI - Effects of fluorescent and nonfluorescent tracing methods on lymphocyte migration in vivo. AB - BACKGROUND: The use of fluorescent dyes to monitor in vivo cellular migration and proliferation has greatly expanded, but little is known about their potential influence on cell migration. METHODS: Adoptive transfer studies of lymphocytes labeled with various dyes were performed, and their in vivo homing was compared with that of coinjected unlabeled control cells. In addition, in vitro migration and binding studies were performed to analyze the various steps of transmigration separately. RESULTS: These data showed that the intracellular fluorescent dyes calcein acetoxymethyl ester, 2',7'-bis-(2-carboxyethyl)-5-(and-6) carboxyfluorescein acetoxymethyl ester, 5-chloromethylfluorescein diacetate, 5 (and-6)-carboxyfluorescein diacetate, succinimidyl ester, and fluorescein isothiocyanate affect in vivo homing of especially B lymphocytes to lymphoid organs, without any direct effect on in vitro chemotactic or adhesive activity. The only label that did not affect migration was the extracellular and nonfluorescent molecule biotin, provided that the labeling was performed at room temperature. Interestingly, by using the highly versatile congenic Ly5.1-Ly5.2 system, we also demonstrated intrinsic differences in lymphocyte migration based on allelic differences. CONCLUSIONS: Our data showed that fluorescent labeling of lymphocytes has a severe effect on their homing capacity in vivo. Labeling of cells with biotin appeared to be a good alternative for this purpose; however, if direct fluorescence is required, the negative effects on cell migration should be considered. PMID- 15351988 TI - Architectural analysis of oral cancer, dysplastic, and normal epithelia. AB - BACKGROUND: We present a novel, automated, and quantitative approach to evaluate local epithelial tissue architecture based on mathematical graph theory. METHODS: Four hundred forty-one images of three diagnostic classes of oral epithelium (normal, dysplastic, and neoplastic) were analysed. The epithelial compartment was partitioned into exclusive areas associated with each nucleus to approach the theoretical cell extents. The spatial arrangement of cells in neighbourhoods of two sizes was characterised by constructing graph networks based on the cell centroids and recording 29 statistical properties. We analysed 104,627 and 67,590 neighbourhoods of diameters 37.5 and 75 microm, respectively. RESULTS: The discrimination power of the architectural descriptors was evaluated by using discriminant analysis. The best neighbourhood discrimination rate was 75% for normal versus carcinoma. For the pooled data, discrimination into three classes based on largest number of neighbourhoods associated with each class was 100% correct. Case-wise, discrimination rates were 67%, 100%, and 80% correct for normal, premalignant, and malignant. When considering two classes, discrimination rates was 89% (normal) and 100% (malignant) correct, with 71% of premalignant cases assigned to the malignant class. CONCLUSIONS: The results indicate that unbiased and reproducible quantification of tissue architectural features is possible and may provide valuable morphological information for diagnostic purposes. PMID- 15351989 TI - Use of SYBR14, 7-amino-actinomycin D, and JC-1 in assessing sperm damage from rats with spinal cord injury. AB - BACKGROUND: Although fluorescent dyes combined with flow cytometry have been used to confirm the viability of sperm in the past, methods to detect damage to spermatozoa following injury have been limited to use of dyes, which are often difficult to adequately compensate for in a single laser system. METHODS: In this article, we present what we believe is a better method to assess damage to sperm secondary to spinal cord injury in an in vivo model, for use with a standard Ar laser and flow cell. In this rat model of spinal cord injury leading to sperm damage, the spinal cords of the rats were injured, but the reproductive organs were not. To understand the origins of sperm injury, and to develop ways to overcome the loss of fertility, we used the viability dye SYBR-14 along with 7 amino actinomycin D to detect apoptosis. Additionally, we used the dye JC-1 to measure the changes in mitochondrial transmembrane potential that accompany the damage. RESULTS: We found that SYBR-14 plus 7-amino actinomycin D was a useful method for quantifying apoptosis, particularly when another dye, such as JC-1, was used simultaneously. By using these dyes in concert with motility studies, we were able to quantify the extent of damage to sperm and correlate it to the decrease in motility of sperm (r(2) = 0.99 for SYBR14 versus motility and r(2) = 0.98 for JC-1 versus motility by regression analysis). CONCLUSIONS: With a method established to measure injury to sperm, we hope to determine which treatment regimens of ones we will test are effective in restoring sperm to a more fertile state, in the future. PMID- 15351990 TI - LAMP-1 and LAMP-2, but not LAMP-3, are reliable markers for activation-induced secretion of human mast cells. AB - BACKGROUND: Mast cells are resident tissue cells that induce anaphylactic reactions by rapidly releasing mediators after antigen-mediated cross-linking of immunoglobulin E receptors. In the similarly active peripheral blood basophilic leukocyte, lysosome-associated membrane protein 3 (LAMP-3; CD63) has been described as an activation marker, but LAMPs have not been investigated in normal tissue mast cells. METHODS: Intra- and extracellular expressions of LAMP-1 (CD107a), LAMP-2 (CD107b), and LAMP-3 (CD63) were analysed by flow cytometry, immunocytochemistry, and functional assays in unstimulated and stimulated leukemic human mast cell line 1 (HMC-1) and skin mast cells. RESULTS: On flow cytometry, all mast cells expressed LAMP-3 at their cell membranes, whereas LAMP 1 and LAMP-2 were barely detectable (HMC-1 cells) or expressed at low levels (<10% of skin mast cells). After fixation and permeabilisation, high intracellular levels of all three LAMPs were noted in both cell types. After stimulation, a rapid translocation of intracellular LAMPs to the cell membrane, with an associated release of histamine, leukotriene C(4) and prostaglandin D(2), was ascertained in skin mast cells only. CONCLUSION: These results show that LAMP 1 and LAMP-2 are activation markers for normal mast cells. The lack of LAMP translocation after activation of leukemic mast cells may be related to maturation or malignancy-associated defects of these cells. PMID- 15351991 TI - B-cell subsets in blood and lymphoid organs in Macaca fascicularis. AB - BACKGROUND: Cynomolgus monkeys (Macaca fascicularis) are widely used animal models in biomedical research. However, the phenotypic characteristics of cynomolgus monkey (CM) B cells in peripheral blood (PB) and lymphoid organs are poorly understood. METHODS: FACS analyses of PB-, spleen-, lymph node (LN)-, and bone marrow (BM)-derived B cells were performed. RESULTS: CM peripheral blood B cells have a smaller fraction of CD27(-) (naive) cells ( approximately 40%), as compared to human blood samples ( approximately 70%). Similar to humans, an early activation marker, CD23, is expressed more on CD27(-) CM naive B cells, as compared to CD27(+) B cells. The mean fraction of B cells exhibiting a memory phenotype is similar to that seen in human blood. Unlike humans, CM blood contains a subset of CD20(++)CD80(+)CD21(-)IgM(+/-)CD27(+)CD19(+)FSC(++)BAFF R(low) B cells that are likely of germinal center origin. Thus, CM blood contains (i) a higher percentage of B cells that express the co-stimulatory molecule CD80, and (ii) a lower fraction of B cells that are CD21(+), as compared to human blood. Due to the relative paucity of information on B-cell subsets in organs of healthy humans, a direct comparison between human and CM lymphoid organ data is limited. The fraction of CD27(+) and CD23(+) B cells appears to be similar, while the fraction of CD80(+) B cells appears to be higher than that seen in human lymphoid organs. CM spleens and to some extent lymph nodes have a distinct subset of CD21(++) cells that are also CD80(+/-)CD23(low)IgM(++)CD27(+/-)FSC(++). This subset is phenotypically similar to the marginal zone B cells present in human spleen and LN samples. We also provide detailed analyses on the fraction of lymphoid organ B cells that express CD21, CD23, CD32, and/or CD80 B-cell markers. CONCLUSIONS: In general, cynomolgus monkey B-cell subsets are similar to those seen in humans, as well as to those seen in other nonhuman primates. However, there are some clear differences between human and cynomolgus monkey B-cell subsets. These findings have direct implications for a variety of in vivo studies in cynomolgus monkeys, ranging from basic research on primate B-cell differentiation to models of infectious diseases and trials of new B-cell targeting therapeutic agents. PMID- 15351992 TI - Salicylic acid-mediated potentiation of Hsp70 induction correlates with reduced apoptosis in tobacco protoplasts. AB - BACKGROUND: Elevated temperatures jeopardize plant disease resistance, as mediated by salicylic acid (SA). SA potentiates heat-induced expression of the 70 kDa heat shock protein (Hsp70) in tomato cells. In mammalian cells, Hsp70 suppresses apoptosis. We hypothesized that potentiation of heat-induced Hsp70 by SA contributes to a reduction in apoptosis in tobacco protoplasts. METHODS: Tobacco protoplasts (Nicotiana tabacum) were exposed to SA (70 microM) at normal temperatures or in combination with heat shock. Hsp70/Hsc70 accumulation and phosphatidylserine (PS) exposure, DNA fragmentation, as well as loss of mitochondrial membrane potential were quantified by flow cytometry. RESULTS AND CONCLUSIONS: SA at normal temperatures did not influence Hsp70/Hsc70 accumulation, but were found to induce apoptosis. In contrast, SA in combination with HS potentiated heat-induced Hsp70/Hsc70 accumulation in tobacco protoplasts that correlated negatively with apoptosis, illustrated by decreased PS exposure and DNA fragmentation and enhanced mitochondrial membrane potential. We propose that this correlation supports a possible role for apoptosis suppression by Hsp70 under elevated temperatures during pathogen infection. PMID- 15351993 TI - Affinity of single S. cerevisiae cells to 2-NBDglucose under changing substrate concentrations. AB - BACKGROUND: Saccharomyces cerevisiae is a widely employed microorganism in biotechnological processes. Since proliferation and product formation depend on the capacity of the cell to access and metabolize a carbon source, a technique was developed to enable for analyzing the S. cerevisiae H155 cells' affinity to extracellular glucose concentrations. METHODS: The fluorescent glucose analogue 2 NBDglucose was employed as a functional parameter to analyze the cells' affinity to glucose. Structural parameters (proliferation, neutral lipid content, granularity, and cell size) were also investigated. Cells were grown both in batches and in chemostat regimes. RESULTS: The 2-NBDglucose uptake in individual cells proceeds in a time- and concentration-dependent manner and is affected by respiratory and respirofermentative modes of growth. The process is inhibited by D-glucose, D-fructose, D-mannose, and sucrose, but not L-glucose, D-galactose or lactose; maltose is a weak inhibitor. The affinity of the individual cells to 2 NBDglucose was found to be high at low extracellular glucose concentrations, and weak at high concentrations. An additional, underlying pattern in the cells' affinity to glucose was detected, illustrated by the recurrent appearance of two subpopulations showing distinctly differing quantities of this substrate. CONCLUSIONS: A multiparameter flow cytometry approach is presented that enables, for the first time, for analysis of the affinity of individual S. cerevisiae cells to glucose. Besides the adjustment of the yeast cell metabolism to extracellular glucose concentrations by altering their affinity to glucose, at least one further mechanism is clearly involved. Two subpopulations of cells were resolved, with different affinities not correlated with other cellular parameters measured. PMID- 15351995 TI - Enantiomeric resolution of 2-arylpropionic acid nonsteroidal anti-inflammatory drugs by capillary electrophoresis: methods and applications. AB - The 2-arylpropionic acids (2-APAs) are an important group of nonsteroidal anti inflammatory drugs. These agents, the majority of which are available as racemates, exhibit stereoselectivity in both their action and disposition. Developments in stereoselective separation science methodology, mainly chromatographic, have facilitated an evaluation of the pharmacological properties of the individual enantiomers of these drugs and contributed to our understanding of both their mode(s) of action and disposition. While a number of electrophoretic techniques, including capillary electrophoresis, capillary electrochromatography and isotachophoresis, have been applied to the stereoselective resolution and stereospecific analysis of these agents using a variety of chiral selectors, e.g., cyclodextrins, oligosaccharides, macrocyclic antibiotics, and proteins, the number of published applications in pharmaceutical and biomedical analysis remains relatively limited. However, the utility of electrophoretic techniques for stereospecific analysis may be illustrated using the 2-APAs as typical examples of chiral acidic pharmaceuticals. Applications include: determination of enantiomeric composition following biosynthetic stereoselective hydrolysis; examination of both achiral and chiral impurity profiles in bulk drugs and formulated products; determination of enantiomeric impurities in both bulk drugs and formulated products; examination of configurational stability following stress testing of formulated products; determination of enantiomeric composition and metabolite profile in biological fluids following administration of the racemates and individual enantiomers. It may be anticipated that future exploitation of electrophoretic approaches to the stereospecific analysis of these agents will result in further contributions to our understanding of their stereoselective biological properties and therapeutic use. PMID- 15351996 TI - Single-isomer sulfated alpha-cyclodextrins for capillary electrophoresis: hexakis(2,3-di-O-methyl-6-O-sulfo)-alpha-cyclodextrin, synthesis, analytical characterization, and initial screening tests. AB - The third, concluding member of the family of single-isomer, fully sulfated alpha cyclodextrins, the sodium salt of hexakis(2,3-di-O-methyl-6-O-sulfo)-alpha cyclodextrin (HxDMS), has been synthesized on the kilogram scale, completing the nine-member array of the single-isomer, 6-O-sulfo CDs now available. HxDMS was tested for the capillary electrophoretic (CE) resolution of the enantiomers of nonelectrolyte, weak acid and weak base analytes contained in our CD screening kit. HxDMS complexed differently with many of the analytes tested than either its larger-ring analogs, heptakis(2,3-di-O-methyl-6-O-sulfo)-beta-CD (HDMS) and octakis(2,3-di-O-methyl-6-O-sulfo)-gamma-CD (ODMS) or its same-ring, but differently substituted analogs, hexakis(6-O-sulfo)-alpha-CD (HxS) and hexakis(2,3-di-O-acetyl-6-O-sulfo)-alpha-CD (HxDAS). For all analytes, the effective mobilities and separation selectivities as a function of the background electrolyte concentration of HxDMS followed the trends that were found for the other single-isomer, 6-O-sulfo CDs. PMID- 15351997 TI - Enantioseparation using sulfated cyclosophoraoses as a novel chiral additive in capillary electrophoresis. AB - Highly sulfated cyclosophoraoses (HS-Cys) were synthesized by the chemical modification of a family of neutral cyclosophoraoses isolated from Rhizobium leguminosarum bv. trifolii. The HS-Cys were then analytically characterized using Fourier transform-infrared spectroscopy and elemental analysis. These HS-Cys were successfully used as a novel chiral additive, in low-pH aqueous background electrolytes, for capillary electrophoretic separation of five basic chiral drugs such as arterenol, atenolol, isoproterenol, propranolol, and metoprolol. PMID- 15351998 TI - Influence of (hydroxy)alkylamino substituents on enantioseparation ability of single-isomer amino-beta-cyclodextrin derivatives in chiral capillary electrophoresis. AB - A family of single-isomer amino-beta-cyclodextrin (amino-beta-CD) derivatives containing an amino or (hydroxy)alkylamino group in one of the primary positions has been synthesized. The steric effect and hydrogen bond forming ability of the different substituents on enantioseparation of acidic enantiomers has been studied by capillary electrophoresis (CE). Three enantiomeric model compounds (mandelic acid, cis-permethrinic acid, and cis-deltamethrinic acid) having significantly different apparent complex stability constants with beta-CD were applied in the experiments. Dependence of separation selectivity, resolution as well as mobility difference on chiral selector concentration (0.1-20 mM, pH 6.0) was investigated. Each amino-beta-CD showed higher enantioselectivity than the native beta-CD. One hydroxyalkyl group attached to the primary amino N-atom significantly increased both the enantioselectivity and the resolution compared to the primary amino-beta-CD, while two hydroxyalkyl moieties decreased them due to the predominance of steric hindrance. The value of the apparent complex stability constants obtained suited well the mobility difference model (by Wren). On the other hand, the optimum selector concentrations calculated according to the model were slightly lower than the experienced concentrations giving the maximum enantioresolution of enantiomers. PMID- 15351999 TI - Evaluation of balhimycin as a chiral selector for enantioresolution by capillary electrophoresis. AB - The glycopeptide antibiotic balhimycin and its haloanalogue bromobalhimycin were evaluated as chiral selectors for enantioresolution by capillary electrophoresis. In order (i) to eliminate the adsorption of the glycopeptide antibiotics on the capillary wall, (ii) to shorten the separation time and (iii) to improve the detection sensitivity, a combined approach of the dynamic surface coating technique, the co-electroosmotic flow electrophoresis technique and the partial filling technique was employed for the enantioresolution of 16 acidic racemates. The effect of experimental parameters (plug length of the partial filling solution containing the chiral selector, selector concentration and buffer pH) on enantiorecognition was investigated. Furthermore, the enantiorecognition ability imparted by balhimycin, bromobalhimycin and vancomycin were compared. For most tested compounds, the highest enantiorecognition was obtained with balhimycin as chiral selector. Only in the case of the enantioresolution of tiaprofenic acid, vancomycin showed a superior enantiorecognition. PMID- 15352000 TI - Effects of partial/asymmetrical filling of micelles and chiral selectors on capillary electrophoresis enantiomeric separation: generation of a gradient. AB - The effect of several experimental parameters on enantiomeric separations in micellar capillary electrophoresis (MCE) was studied. A model separation system was tested. It was composed of an acidic phosphate buffer with sodium dodecyl sulfate (SDS) and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as the chiral selector. A substituted angelicin was used as a chiral analyte. Changes in the concentration of SDS micelles/SDS monomers in the presence of HP-beta-CD and their impact on the enantioselective separation were investigated. Variation of the composition of electrolytes in the individual compartments of the separation system (inlet vial, capillary, and outlet vial) affected both the migration times and the resolution of the enantiomers. Current vs. time dependencies also were monitored during the separations. A mathematical model of electromigration in micellar systems with chiral selector present was proposed and a computer simulation was used to explain the observed phenomena and to confirm the generation of a CD/SDS-micelle concentration gradient under certain experimental conditions. This is the reported first attempt of a computer simulation of the complex, dynamic chiral environment of the CD-SDS-MCE system. PMID- 15352001 TI - Enantiomeric separation of basic compounds using heptakis(2,3-di-O-methyl-6-O sulfo)-beta-cyclodextrin in combination with potassium camphorsulfonate in nonaqueous capillary electrophoresis: optimization by means of an experimental design. AB - The enantiomeric separation of a series of basic pharmaceuticals (beta-blockers, local anesthetics, sympathomimetics) has been investigated in nonaqueous capillary electrophoresis (NACE) systems using heptakis(2,3-di-O-methyl-6-O sulfo)-beta-cyclodextrin (HDMS-beta-CD) in combination with potassium camphorsulfonate (camphorSO3-). For this purpose, a face-centered central composite design with 11 experimental points was applied. The effect of the concentrations of HDMS-beta-CD and camphorSO3- on enantioresolution was statistically evaluated and depended largely on the considered analyte. The presence of camphorSO3- was found to be particularly useful for the enantioseparation of compounds with high affinity for the anionic CD. CamphorSO3- seems to act as a competitor, reducing the affinity for the CD, probably by ion pair formation with these analytes. For compounds with lower affinity for HDMS beta-CD, the combination of camphorSO3- and the CD appeared to have a favorable effect on enantioresolution only if the optimal CD concentration could be reached. On the other hand, for compounds characterized by a very low affinity for the anionic CD, the association of camphorSO3- and HDMS-beta-CD is always unfavorable. Finally, experimental conditions were selected by means of the multivariate approach in order to obtain the highest resolution (Rs) value for each studied compound. PMID- 15352002 TI - Comparative study on the chiral separation of phenyl alcohols by capillary electrophoresis and liquid chromatography. AB - Structurally related phenyl alcohols were separated by capillary electrophoresis and liquid chromatography. A statistical experimental design was used in order to optimize the main electrophoretic parameters such as pH, concentration of selector and separation voltage in capillary electrophoresis (CE). Response surfaces were derived using the mathematical model and used for a selection of the optimal experimental conditions. Concentration of the chiral selector, the distance between the aromatic group and asymmetric center of the analytes, were identified as the factors influencing the complexation, selectivity and resolution. Experiments were also performed by high-performance liquid chromatography (HPLC) and the results of CE and HPLC were compared. PMID- 15352003 TI - Enantioselectivity of structurally modified poly(sodium undecenoyl-L-leucinate) by insertion of Triton X-102 surfactant molecules. AB - The effectiveness of Triton X-102 (TX-102), as a structural modifier of the polymeric surfactant sodium undecanoyl-L-leucinate (L-SUL) was investigated for enhanced enantiomeric recognition of various chiral compounds in micellar electrokinetic chromatography (MEKC). Increasing concentrations of TX-102 were separately added into the micellar solutions of L-SUL and then polymerized to form poly-L-SUL. The resulting polymers were purified by use of 3500 molecular weight-cutoff (MWCO) dialysis membranes. Fluorescence and pulsed field gradient nuclear magnetic resonance (PFG-NMR) techniques were used to elucidate the structural effects of TX-102 on poly-L-SUL. Evaluation of data from fluorescence measurements suggested an increase in polarity with increasing concentration of TX-102. However, the polarity decreased at higher concentrations of TX-102. Evaluation of data from PFG-NMR suggested an increase in hydrodynamic radius upon increasing the concentration of TX-102. The racemates of coumarinic and phenythiohydantoin amino acid derivatives, and pindolol were used as test analytes in MEKC. A notable increase in resolution and capacity factors of the test analytes was observed when the modified poly-L-SUL was used in MEKC measurements. Examination of the data obtained from fluorescence, PFG-NMR, and MEKC suggests a strong correlation between the polarity and the hydrodynamic radii of TX-102 modified micelles and the enantiomeric resolution of the test analytes. PMID- 15352004 TI - Enantiomeric separation of neutral hydrophobic dihydrofuroflavones by cyclodextrin-modified micellar capillary electrophoresis. AB - The enantiomeric separations of several very hydrophobic dihydrofuroflavones were performed and optimized using cyclodextrin-modified micellar capillary electrophoresis. Overall, the greatest enantiomeric peak-to-peak separations for the greatest number of flavones were obtained with hydroxypropyl-gamma cyclodextrin. The effects of cyclodextrin and sodium dodecyl sulfate concentration and pH were examined in order to optimize the separation conditions. The ratio of surfactant-to-cyclodextrin concentration affected the chiral discrimination of the system significantly, with increases in the derivatized cyclodextrin concentration generally enhancing resolution. Higher efficiencies were obtained with lower concentrations of surfactant and cyclodextrin, although enantioseparation optimization often required higher concentrations to be used. A highly acidic pH was necessary to effectively suppress the electroosmotic flow when operating in the reversed polarity mode. Experiments utilizing the normal polarity mode and higher pH produced no separations. PMID- 15352005 TI - Chiral separation of N-imidazole derivatives, aromatase inhibitors, by cyclodextrin-capillary zone electrophoresis. Mechanism of enantioselective recognition. AB - Baseline separation of ten new, substituted [1-(imidazo-1-yl)-1-phenylmethyl)] benzothiazolinone and benzoxazolinone derivatives with one chiral center was achieved using cyclodextrin-capillary zone electrophoresis (CD-CZE). A method for the enantiomeric resolution of these compounds was developed using neutral CDs (native alpha-, beta-, gamma-CDs or alpha-, beta-, gamma-hydroxypropyl (HP)-CDs) as chiral selectors. Operational parameters including the nature and concentration of the chiral selectors, pH, ionic strength, organic modifiers, temperature, and applied voltage were investigated. The use of neutral CDs provides enantiomeric resolution by inclusion of compounds in the CD cavity. The HP-alpha-CD and HP-beta-CD were found to be the most effective complexing agents and allowed efficient enantiomeric resolutions. Optimal separation of N-imidazole derivatives was obtained using 50 mM phosphate buffer at pH 2.5 containing either HP-alpha-CD or HP-beta-CD (7.5-12.5 mM) at 25 degrees C, with an applied field of 0.50 kV.cm(-1) giving resolution factors Rs superior to 1.70 with migration times of the second enantiomer less than 13 min. The same enantiomer migration order observed for all molecules can be related to a close interaction mechanism with CDs. The influence of structural features of the solutes on Rs and tm was studied. The lipophilic character (log kw) of the solutes and the apparent and averaged association constants of inclusion complexes for four compounds with the six different CDs led us to rationalize the enantioseparation mechanisms. The conclusions were corroborated with reversed-phase high-performance liquid chromatography (HPLC) on chiral stationary phases (CSPs) based on CDs. PMID- 15352006 TI - Chiral capillary electrophoresis applied to the determination of phenylglycidol enantiomers obtained from cinnamyl alcohol by asymmetric epoxidation using new titanium(IV) alkoxide compounds as catalysts. AB - A capillary electrophoresis method for the simultaneous determination of phenylglycidol enantiomers in the presence of an excess of cinnamyl alcohol was developed. The effects of the nature, pH and concentration of the buffer, the nature and concentration of chiral selector, the addition of methanol or acetonitrile, and the capillary temperature on the chiral resolution of phenylglycidol enantiomers were studied. Separations were achieved using 20 mM succinylated beta-cyclodextrin dissolved in a 10 mM borate buffer (pH 10.0). Chiral resolution for the phenylglycidol enantiomers in the optimized electrophoretic conditions was higher than 2.0 with an analysis time less than 7 min. The method developed was validated in terms of selectivity, linearity, precision (instrumental repeatability, method repeatability, intermediate precision), the limits of detection and quantitation, and accuracy. Limits of detection of 6.5 mg/L and 8.3 mg/L for (2S,3S)-(-)-3-phenylglycidol ((S,S)-PG) and (2R,3R)-(+)-3-phenylglycidol ((R,R)-PG), respectively, were obtained. The method was applied to study the asymmetric epoxidation of cinnamyl alcohol with titanium(IV) alkoxide compounds as catalysts in order to evaluate their catalytic activity and stereoselectivity of the epoxidation processes. PMID- 15352007 TI - Enantioseparation of aromatic amino acids and amino acid esters by capillary electrophoresis with crown ether and prediction of enantiomer migration orders by a three-dimensional quantitative structure-property relationship/comparative field analysis model. AB - The complete enantioseparations of eight aromatic amino acids and four alkyl esters of 2-phenylglycine were achieved by chiral capillary electrophoresis employing 20 mM Tris-citric acid background electrolyte (pH 2.50) containing 5.0 mM of (+)-18-crown-6-tetracarboxylic acid. The relative migration times (RMTs) to that of 6-aminonicotinic acid were characteristic of each enantiomer with good within-day precisions (% relative standard deviation (RSD) < or = 2.0). Quantitative structure-property relationship (QSPR) modeling based on comparative molecular field analysis (CoMFA) was performed to investigate the correlation between the molecular field descriptor values of each enantiomer studied as analyte and its RMT. The resulting CoMFA model allowed reliable prediction for the RMT values (q2= 0.406, r2 = 0.996), thus being expected to become a valuable tool to predict enantiomer migration orders (EMOs) of amino acids and amines whose pure enantiomers are unavailable. The CoMFA steric fields supported the well-established chiral recognition mechanism based on molecular interaction between chiral selector (+)-18-crown-6-tetracarboxylic acid and amino acid enantiomers. PMID- 15352008 TI - Ultrashort partial-filling technique in capillary electrophoresis for infinite resolution of tramadol enantiomers and its metabolites with highly sulfated cyclodextrins. AB - The possibility to enhance resolution to infinite value in chiral capillary electrophoresis is attained as soon as the apparent mobility of one enantiomer becomes opposite to the other. This could be achieved on the basis of the carrier ability of multiple charged chiral selectors such as highly sulfated cyclodextrin (HS-CD). With tramadol and its phase I metabolites selected as model compounds, the HS-gamma-CD was found to be the most appropriate chiral selector. The CD concentration was determined where one enantiomer still migrated as a cation while the other migrated in the opposite side. Besides the chiral selector concentration, secondary parameters such as buffer concentration appeared to be critical to reach infinite resolution. The latter was achieved with partial filling technique using ultrashort separation zones (a few mm). In order to better understand the interaction mechanism between the selected CD and the analytes, the classical affinity capillary electrophoresis method, although not fully satisfactory because of ionic strength variations within a series of mobility shift measurements, was applied to estimate complexation constants and complex mobilities. The results obtained point to the prevailing role of complex mobility differences in the enantioselectivity mechanism. PMID- 15352009 TI - A screening strategy for the development of enantiomeric separation methods in capillary electrophoresis. AB - Method development of enantiomeric separations in capillary electrophoresis (CE) is a time-consuming task, since finding the appropriate chiral selector is usually a "trial and error" process. It is impossible to predict the selectivity of a selector towards a certain enantiomer. Therefore, the affinity of all selectors has to be examined one at a time. In order to speed up this process, a strategy is proposed based on simple experimental design methodology. The approach includes first a screening in function of the pH to determine the optimal migration conditions followed by a selection of the right chiral selector by means of Taguchi designs. In the approach several variables, such as the type and concentration of cyclodextrin, the concentration of buffer electrolyte, and the percentage of organic modifier, are varied simultaneously to find initial separation conditions rapidly. The resulting initial separation conditions can be optimized in further steps to be more reproducible. We discuss the results of the approach when applied on a number of selected compounds that are recently in development at Johnson & Johnson--Pharmaceutical Research and Development. Parameters, such as quality of the separation and analysis time, are evaluated to determine initial separation conditions for each compound. PMID- 15352010 TI - Enantioseparations of hydrobenzoin and structurally related compounds in capillary zone electrophoresis using heptakis(2,3-dihydroxy-6-O-sulfo)-beta cyclodextrin as chiral selector and enantiomer migration reversal of hydrobenzoin with a dual cyclodextrin system in the presence of borate complexation. AB - We investigated the enantioseparations of racemic hydrobenzoin, together with benzoin and benzoin methyl ether, in capillary electrophoresis (CE) using the single-isomer heptakis(2,3-dihydroxy-6-O-sulfo)-beta-cyclodextrin (SI-S-beta-CD) as a chiral selector in the presence and absence of borate complexation and enantiomer migration reversal of hydrobenzoin with a dual CD system consisting of SI-S-beta-CD and beta-CD in the presence of borate complexation at pH 9.0 in a borate buffer. The enantioselectivity of hydrobenzoin increased remarkably with increasing SI-S-beta-CD concentration and the enantioseparation depended on CD complexation between hydrobenzoin-borate and SI-S-beta-CD. The (S,S)-enantiomer of hydrobenzoin-borate complexes interacted more strongly than the (R,R) enantiomer with SI-S-beta-CD. The enantiomers of hydrobenzoin could be baseline resolved in the presence of SI-S-beta-CD at a concentration as low as 0.1% w/v, whereas the three test analytes were simultaneously enantioseparated with addition of 0.3% w/v SI-S-beta-CD or at concentrations >2.0% w/v in a borate buffer and 0.5% w/v in a phosphate background electrolyte at pH 9.0. Compared with the results obtained previously using randomly sulfated beta-CD (MI-S-beta CD) in a borate buffer, enantioseparation of these three benzoin compounds is more advantageously aided by SI-S-beta-CD as the chiral selector. The enantioselectivity of hydrobenzoin depended greatly on the degree of substitution of sulfated beta-CD. Moreover, binding constants of the enantiomers of benzoin compounds to SI-S-beta-CD and those of hydrobenzoin-borate complexes to SI-S-beta CD were evaluated for a better understanding of the role of CD complexation in the enantioseparation and chiral recognition. Enantiomer migration reversal of hydrobenzoin could be observed by varying the concentration of beta-CD, while keeping SI-S-beta-CD at a relatively low concentration. SI-S-beta-CD and beta-CD showed the same chiral recognition pattern but they exhibited opposite effects on the mobility of the enantiomers. PMID- 15352011 TI - Heptakis(6-amino-6-deoxy)-beta-cyclodextrin as a chiral selector for the separation of anionic analyte enantiomers by capillary electrophoresis. AB - A hepta-substituted beta-cyclodextrin bearing seven amino groups, heptakis(6 amino-6-deoxy)-beta-cyclodextrin (per-6-NH2-beta-CD) was successfully used as a chiral selector for the enantioseparation of different anionic analytes. The running buffer pH and chiral selector concentration were the studied parameters crucial in achieving the maximum possible enantioresolution. Enantiomeric separation of a mixture of seven carboxybenzyl-amino acids was achieved in 24 min. Excellent resolution was obtained for carboxybenzyl-tryptophan (Rs = 11.2). PMID- 15352012 TI - Chiral capillary electrophoresis: facts and fiction on the reproducibility of resolution with randomly substituted cyclodextrins. AB - Comparative enantioseparation of the enantiomers of 1,1'-binaphthyl-2,2'-diyl hydrogen phosphate was performed with cyclodextrin (CD)-modified capillary electrophoresis (CE). Two single isomers, beta-CD, heptakis(2,3,6-tri-O-methyl) beta-CD (TM-beta-CD), and heptakis(2,6-di-O-methyl)-beta-CD (DM-beta-CD) of 98% purity as well as heptakis(2,3-di-O-acetyl)-beta-CD were used and compared in terms of resolution power to randomly methylated and corresponding acetylated beta-CDs, which were synthesized in our laboratory. The methylated ones were characterized by means of matrix-assisted laser desorption/ionization-time of flight-(MALDI-TOF) mass spectrometry. By testing defined mixtures of single isomers and comparing their resolution power to randomly substituted CDs of similar degree of substitution we could show, that a simple characterization by the average molecular degree of substitution (DS) is not sufficient. In order to get reproducible results, a clearly defined substitution pattern is necessary, which is not given using randomly substituted CDs. Taken together, a validation of a chiral separation with "undefined" CD derivatives is almost impossible. PMID- 15352013 TI - Enantioseparations by capillary electro-chromatography: differences exhibited by normal- and reversed-phase versions of polysaccharide stationary phases. AB - The influence of using normal-phase and reversed-phase versions of four commercial polysaccharide stationary phases on chiral separations was investigated with capillary electrochromatography (CEC). Both versions of the stationary phases, Chiralcel OD, OJ, and Chiralpak AD, AS were tested for the separation of two basic, two acidic, a bifunctional, and a neutral compound. Different background electrolytes were used, two at low pH for the acid, bifunctional and neutral substances, and three at high pH for the basic, bifunctional and neutral ones. This setup allowed evaluating differences between both stationary-phase versions and between mobile-phase compositions on a chiral separation. Duplicate CEC columns of each stationary phase were in-house prepared and tested, giving information about the intercolumn reproducibility. In general, reversed-phase versions of the current commercial polysaccharide stationary phases are found to be best for reversed-phase CEC, even though at high pH no significant differences were seen between both versions. Most differences were observed at low pH. For acidic compounds, it was seen that an ammonium formate electrolyte performed best, which is also an excellent electrolyte if coupling with mass spectrometry is desired. For basic, bifunctional and neutral compounds, no significant differences between the three tested electrolytes were observed at high pH. Here, a phosphate buffer is preferred as electrolyte because of its buffering capacities. However, if coupling to mass spectrometry is wanted, the more volatile ammonium bicarbonate electrolyte can be used as an alternative. PMID- 15352014 TI - Preparation of a positively charged cellulose derivative chiral stationary phase with copolymerization reaction for capillary electrochromatographic separation of enantiomers. AB - A positively charged chiral stationary phase (CSP) was prepared by chemically immobilizing cellulose 3,5-dimethylphenylcarbamate onto methacryloyldiethylenetriaminopropylated silica (MCDEAPS) via a radical copolymerization reaction. The prepared CSP was evaluated for enantiomer separation in nonaqueous capillary electrochromatography (CEC). Electroosmotic flow (EOF) generated on the prepared CSP could be significantly improved with introduction of positive charges into the CSP, and separation of enantiomers in CEC has been achieved with mobile phases of ethanol and hexane-ethanol, respectively. In addition, we investigated the solvent versatility of the immobilized CSP on enantioseparations in CEC and capillary liquid chromatography (CLC) due to the elimination of dissolution of chiral selector in a number of solvents. Chiral resolution of some enantiomers was improved by adopting tetrahydrofuran (THF) and chloroform as mobile-phase modifiers, respectively. PMID- 15352015 TI - A simplified synthesis of polymeric nonparticulate stationary phases with macrocyclic antibiotic as chiral selector for capillary electrochromatography. AB - A simplified approach to synthesize nonparticulate (continuous or monolithic) beds with embedded vancomycin chiral selectors for capillary electrochromatography is proposed. In the present approach, N,N' diallyltartardiamide monomer with diol functionality is used, which can be readily converted to aldehyde groups via periodate treatment. Parallel to the activation of the polymeric matrix for covalent attachment of vancomycin, the periodate treatment has shown secondary effects on the polymeric bed morphology, namely the increase of the average pore size and porosity of the skeleton. Inversed size-exclusion chromatography was applied to characterize porosimetric properties of the capillary columns before and after the periodate treatment. Electroosmotic and enantioselective properties of the nonparticulate beds synthesized are presented. The approach is of more general interest attaching different affinity groups to the polymeric matrix and/or enhancing the accessibility to the active sites, for instance, in the molecular imprinting technique. PMID- 15352016 TI - Determination of the chiral and achiral related substances of methotrexate by cyclodextrin-modified micellar electrokinetic chromatography. AB - A cyclodextrin-modified micellar electrokinetic chromatographic (CD-MEKC) method for the determination of the most important potential impurities of methotrexate (MTX): 2,4-diamino-6-(hydroxymethyl)pteridine, aminopterine hydrate, 4-[N-(2 amino-4-hydroxy-6-pteridinylmethyl)-N-methylamino] benzoic acid, 4-[N-(2,4 diamino-6-pteridinylmethyl)-N-methylamino] benzoic acid, and the distomer D-MTX is presented. The MEKC separation of these compounds was optimized by applying a step-by-step approach. The addition of beta-CD to a conventional MEKC system, based on sodium dodecyl sulfate (SDS) as surfactant, showed to be essential for the enantioresolution of racemic MTX as well as for the separation of the achiral impurities. To achieve high-resolution factor between the peaks adjacent to the main component (L-MTX), as required in the analysis of related impurities, the separation conditions were stressed; in particular, the addition of methanol to the CD-MEKC system resulted in a very effective choice. Under the optimized final conditions (100 mM SDS and 45 mM beta-CD in a mixture of 50 mM borate buffer, pH 9.30-methanol (75:25 v/v)), the method was validated showing a general adequate accuracy (93-106% recovery) in the determination of L-MTX related substances at the impurity level of 0.12% w/w with a relative standard deviation (RSD)% lower than 8% (n = 4). The method was successfully applied to the analysis of pharmaceuticals (tablets and injections) which showed to contain the distomer D MTX as major impurity and aminopterine hydrate as a further related substance in the commercial tablets. PMID- 15352017 TI - Influence of methanol on the enantioresolution of antihistamines with carboxymethyl-beta-cyclodextrin in capillary electrophoresis. AB - According to the model of Wren and Rowe, the separation between two enantiomers in capillary electrophoresis (CE) decreases if an organic modifier is added to the run buffer containing a neutral cyclodextrin (CD) in a concentration below its optimal value in a solvent-free system. In previous work, however, it was observed that the addition of methanol to the background electrolyte (BGE) containing not charged carboxymethyl-beta-CD in a concentration below its optimal value, increased the enantioresolution of dimetindene maleate. The enantioresolution decreased when other organic modifiers (ethanol, isopropanol or acetonitrile) were added and/or when other neutral (beta-CD, hydroxypropyl-beta CD) or chargeable (carboxyethyl-beta- and succinyl-beta-CD) CDs were used. In this CE study further attempts are made to elucidate the observed phenomena through investigating other basic drugs. The effect of organic modifier and CD concentration on the enantioseparation was studied by means of central composite designs. It is shown that obtaining this increase in enantioresolution depends upon the type of CD, the type of organic modifier, and the structure of the analytes. It was also observed that small differences in the structure of the analytes or the CD could have an influence on the enantioresolution. The addition of methanol also resulted in different effects on the resolution of closely related analytes. PMID- 15352018 TI - Enantioselective analysis of primaquine and its metabolite carboxyprimaquine by capillary electrophoresis. AB - An enantioselective capillary electrophoresis method for the simultaneous determination of primaquine (PQ) and carboxyprimaquine (CPQ) in rat liver mitochondrial fraction, suitable for in vitro metabolism studies is presented. The drug and metabolite were extracted by liquid-liquid extraction using ethyl ether. The enantiomers were resolved in a fused-silica capillary, 50 microm inside diameter (ID) and 24 cm of effective length, using an electrolyte solution consisting of a 20 mmol/L sodium phosphate solution, pH 3.0, and 10% w/v maltodextrin. Hydrodynamic sample injection was used with a 10 s injection time at 50 mbar pressure. The applied voltage was 22 kV and the capillary temperature was controlled at 20 degrees C. Detection was carried out at 264 nm. Under these conditions, the enantiomeric fractions of the drug and of its metabolite were analyzed within 6 min. The extraction procedure was efficient in removing endogenous interferents and low values (<10%) for the coefficients of variation and deviation from theoretical values were demonstrated for both within-day and between-day assays. The method described allows the determination of PQ and CPQ enantiomers as low as 100 and 40 ng/mL, respectively. After validation, the method was used for an in vitro metabolism study of PQ. The results showed that the enantiomer (-)-PQ was preferentially metabolized to (-)-CPQ. PMID- 15352019 TI - Development and validation of a capillary electrophoresis method for the enantiomeric purity determination of SLV307, a basic potential antipsychotic compound. AB - A capillary electrophoresis method for the enantiomeric purity determination of SLV307, a basic potential antipsychotic compound, has been developed and validated. It is shown that the presence of the eutomer in the sample solution may have a significant effect on the peak shape of the distomer, due to electrodispersion. The method is shown to give good performance with respect to the validation parameters specificity, linearity, accuracy, precision, determination limits, stability of solutions, and robustness of extraction. PMID- 15352020 TI - Evaluation of chiral purity for a substituted imidazole p38 MAP kinase inhibitor and its intermediates using a single chiral capillary electrophoresis method. AB - The chiral separation of a substituted imidazole p38 MAP kinase inhibitor and its intermediates was investigated using capillary electrophoresis (CE) with various sulfated cyclodextrins. After initial screens, a single CE chiral method with a randomly sulfated beta-CD was selected for the evaluation of chiral purity for all three compounds. Operational parameters, such as the concentration of the chiral selectors, background electrolyte (or mobile phase) pH, organic modifiers, and temperature were varied in order to achieve an optimized method. The optimal method was validated in terms of linearity, sensitivity, precision, ruggedness, and specificity. PMID- 15352021 TI - Assessment of the N-oxidation of deprenyl, methamphetamine, and amphetamine enantiomers by chiral capillary electrophoresis: an in vitro metabolism study. AB - A chiral capillary electrophoresis method using hydroxypropyl-beta-cyclodextrin as chiral selector was developed and validated for the quantification of the N oxygenated metabolites of deprenyl, methamphetamine, and amphetamine enantiomers, formed in vitro. The influence of various parameters (selector concentration, buffer pH, temperature, polymer additive, etc.) on the simultaneous separation of the optical isomers of the parent drugs and their metabolites has been evaluated. The buffer pH had the greatest impact on the separation selectivity of the N oxygenated compounds. Linear calibration curves were obtained over the concentration range of 2.5-50 microM for the enantiomers of amphetamine hydroxylamine, methamphetamine-hydroxylamine, and deprenyl-N-oxide. The inter- and intra-assay precision and accuracy varied by less than 15% for all analytes at concentrations of 5, 10, and 30 microM, and less than 20% at the lower limit of quantitation (2.5 microM). The sample extraction recovery ranged between 109 and 129% at the three concentration levels. The drug enantiomers were incubated with recombinant human flavin-containing monooxygenase enzymes (FMO3 and FMO1), and human liver microsomes, respectively. The enantioselectivity of the substrate preference, as well as the stereoselective formation of the new chiral center upon the oxidation of the prochiral tertiary nitrogen of deprenyl were assessed. FMO1, the extrahepatic form of the enzyme in man, was shown to be more active in the N-oxygenation of both deprenyl and methamphetamine isomers than FMO3. Deprenyl enantiomers and S-methamphetamine were substrates of human recombinant FMO3. Conversion of amphetamine to its hydroxylamine derivative could not be observed on incubation with either FMO1 or FMO3. Formation of the new chiral center on the nitrogen, during N-oxidation of the tertiary amine deprenyl, was found stereoselective. The two FMO isoforms have shown opposite preference in the formation of this chiral center. Methamphetamine-hydroxylamine formed from methamphetamine was further transformed by FMO, amphetamine-hydroxylamine was identified as the product of a demethylation reaction. PMID- 15352022 TI - Optimization and validation of an enantioselective method for a chiral drug with eight stereo-isomers in capillary electrophoresis. AB - Highly selective capillary electrophoresis (CE) screening methods were applied to find a satisfactory separation of a chiral drug with eight stereoisomeric compounds. The initial separation conditions were further optimized using response surface modelling by applying a Box-Behnken experimental design. This approach resulted in a rapid and efficient optimization of the buffer concentration, the concentration of two cyclodextrins, and the run voltage, in order to obtain final separation conditions of the method. Further optimization and validation of the system in terms of sensitivity and robustness resulted in a method that is suitable for quality control release purposes. PMID- 15352023 TI - Application of stepwise discriminant analysis to classify commercial orange juices using chiral micellar electrokinetic chromatography-laser induced fluorescence data of amino acids. AB - The use of chiral amino acids content and stepwise discriminant analysis to classify three types of commercial orange juices (i.e., nectars, orange juices reconstituted from concentrates, and pasteurized orange juices not from concentrates) is presented. Micellar electrokinetic chromatography with laser induced fluorescence (MEKC-LIF) and beta-cyclodextrins are used to determine L- and D-amino acids previously derivatized with fluorescein isothiocyanate (FITC). This chiral MEKC-LIF procedure is easy to implement and provides information about the main amino acids content in orange juices (i.e., L-proline; L-aspartic acid, D-Asp, L-serine, L-asparagine, L-glutamic acid, D-Glu, L-alanine, L .arginine, D-Arg, and the non-chiral gamma-amino-n-butyric acid (GABA), i.e., gamma-aminobutyric acid). From these results, it is clearly demonstrated that some D-amino acids occur naturally in orange juices. Application of stepwise discriminant analysis to 26 standard samples showed that the amino acids L-Arg, L Asp and GABA were the most important variables to differentiate the three groups of samples. With these three selected amino acids a 100% correct classification of the samples was obtained either by standard or by leave-one-out cross validation procedures. These classification functions based on the content in L Arg, L-Asp and GABA were also applied to nine test samples and provided an adequate classification and/or interesting information on these samples. It is concluded that chiral MEKC-LIF analysis of amino acids and stepwise discriminant analysis can be used as a consistent procedure to classify commercial orange juices providing useful information about their quality and processing. To our knowledge, this is the first report about the combined use of chiral capillary electrophoresis and discriminant techniques to classify foods. PMID- 15352024 TI - Direct enantioseparation of catechin and epicatechin in tea drinks by 6-O-alpha-D glucosyl-beta-cyclodextrin-modified micellar electrokinetic chromatography. AB - Cyclodextrin-modified micellar electrokinetic chromatography was applied to the enantioseparation of catechin and epicatechin using 6-O-alpha-D-glucosyl-beta cyclodextrin together with sodium dodecyl sulfate and borate-phosphate buffer. Factors affecting chiral resolution and migration time of catechin and epicatechin were studied. The optimum running conditions were found to be 200 mM borate-20 mM phosphate buffer (pH 6.4) containing 25 mM 6-O-alpha-D-glucosyl-beta cyclodextrin and 240 mM sodium dodecyl sulfate with an effective voltage of +25 kV at 20 degrees C using direct detection at 210 nm. Under these conditions, the resolution (Rs) of racemic catechin and epicatechin were 4.15 and 1.92, respectively. With this system, catechin and epicatechin enantiomers along with other four catechins ((-)-catechin gallate, (-)-epicatechin gallate, (-) epigallocatechin, (-)-epigallocatechin gallate) and caffeine in tea samples were analyzed successfully. The difference of migration time between catechin and epicatechin is discussed. PMID- 15352027 TI - Histone deacetylase inhibitors: understanding a new wave of anticancer agents. AB - Cancer is as much an epigenetic disease as it is a genetic and cytogenetic disease. The discovery that drastic changes in DNA methylation and histone modifications are commonly found in human tumors has inspired various laboratories and pharmaceutical companies to develop and study epigenetic drugs. One of the most promising groups of agents is the inhibitors of histone deacetylases (HDACs), which have different biochemical and biologic properties but have a single common activity: induction of acetylation in histones, the key proteins in nucleosome and chromatin structure. One of the main mechanisms of action of HDAC inhibitors is the transcriptional reactivation of dormant tumor suppressor genes, such as p21WAF1. However, their pleiotropic nature leaves open the possibility that their well-known differentiation, cell-cycle arrest and apoptotic properties are also involved in other functions associated with HDAC inhibition. Many phase I clinical trials indicate that HDAC inhibitors appear to be well-tolerated drugs. Thus, the field is ready for rigorous biologic and clinical scrutiny to validate the therapeutic potential of these drugs. Our current data indicate that the use of HDAC inhibitors, probably in association with classical chemotherapy drugs or in combination with DNA-demethylating agents, could be promising for cancer patients. PMID- 15352028 TI - Dibenzo[A,L]pyrene-induced genotoxic and carcinogenic responses are dramatically suppressed in aryl hydrocarbon receptor-deficient mice. AB - Dibenzo[a,l]pyrene (DB[a,l]P), a notorious air pollutant, is the most powerful carcinogenic polycyclic aromatic hydrocarbon (PAH) ever tested. Although the carcinogenicity of PAH may be primarily mediated by the aryl hydrocarbon receptor (AhR), the in vivo role of AhR in skin carcinogenesis remains to be defined. In this context, we investigated the genotoxic and carcinogenic responses of the AhR deficient mouse skin to DB[a,l]P. A single painting resulted in a striking epidermal hyperplasia in AhR+/+ mice but not in AhR-/- mice. Bromodeoxyuridine labeling index and accumulation of p53 protein in epidermal cells of AhR+/+ mice were 8- and 33-fold higher than those of AhR-/- mice, respectively. 32P Postlabeling assay for DB[a,l]P-DNA adducts displayed a 2-fold increase in the AhR+/+ mouse skin. After DB[a,l]P exposure, AhR-/- mice arranged a nearly 60% reduction in the induction of epidermal cytochrome P450 (CYP)1A1, but CYP1B1 was constitutively expressed in both genotypes of mice, irrespective of DB[a,l]P treatment. As compared with AhR+/+ mice, AhR-/- mice had both significantly lower incidence (100% vs. 33%) and multiplicity (2.7 vs. 0.46) of skin tumors by the complete carcinogenesis study. These observations indicate that a reduced tumor yield in AhR-/- mice may be secondary to reduction of inducible CYP1A1 activation and subsequent DNA adduction. It is evident from our continuous work that although AhR is likely to play a central role in epidermal proliferation and possibly neoplastic transformation, the relative importance of AhR for carcinogenesis may be different among PAH examined. PMID- 15352029 TI - Selenoprotein P, as a predictor for evaluating gemcitabine resistance in human pancreatic cancer cells. AB - Gemcitabine is a new standard chemotherapeutic agent used in the treatment of pancreatic cancer, but the mechanisms of gemcitabine sensitivity are still controversial. In our study to determine a mechanism that regulates gemcitabine sensitivity, we carried out molecular analysis on the susceptibility of the pancreatic cancer cells. Using a gemcitabine-sensitive pancreatic cancer cell line KLM1, we established a resistant cell line KLM1-R exhibiting a 20-fold IC50 value (the concentration of gemcitabine causing 50% growth inhibition). Microarray analysis of genes showed specific expression of selenoprotein P, one of the anti-oxidants, in the KLM1-R cell line but not in the KLM1 cell line. Administration of selenoprotein P inhibited the gemcitabine-induced cytotoxicity in the pancreatic cell lines. The levels of intracellular reactive oxygen species (ROS) were increased in the KLM1 cells by gemcitabine, but selenoprotein P suppressed the gemcitabine-induced ROS levels. Furthermore interferon-gamma suppressed the expression of selenoprotein P mRNA and increased intracellular ROS level, leading to the recovery of the gemcitabine sensitivity in KLM1-R. These results suggest a novel mechanism that selenoprotein P reduces the intracellular ROS levels, resulting in the insusceptibility to gemcitabine. PMID- 15352030 TI - Enhanced cathepsin L expression is mediated by different Ras effector pathways in fibroblasts and epithelial cells. AB - Ras expression induces increased expression and altered targeting of lysosomal proteases in multiple cell types, but the specific downstream cytoplasmic signaling pathways mediating these changes have not been identified. In this study, we compared the involvement of 3 major Ras effectors, Raf, phosphatidylinositol 3-kinase (PI3K) and Ral guanine nucleotide exchange factor (RalGEF) in the Ras-mediated alteration of lysosomal protease protein expression and targeting in rat 208F fibroblasts and rat ovarian surface epithelial (ROSE) cells. Effector domain mutants of Ras, constitutively activated variants of Raf, PI3K and RalGEF and pharmacologic inhibitors of MEK and PI3K were utilized to determine the role of these downstream pathways in mediating fibroblast transformation and lysosomal protease regulation in the fibroblasts and epithelial cells. We found that Raf activation of the ERK mitogen-activated protein kinase pathway alone was sufficient to cause morphologic and growth transformation of the fibroblasts and was necessary and sufficient to alter cathepsin L expression and targeting. In contrast, transformation and upregulation of cathepsin L expression in the epithelial cells required the activity of all 3 Ras effectors. Increased protease secretion from the epithelial cells was not observed on ectopic expression of Ras, as it was from the fibroblasts, consistent with the utilization of different signaling pathways in the 2 cell types. In neither cell type did Ras expression increase the expression, processing or secretion of 2 other major lysosomal proteases, cathepsin B and cathepsin D. Thus, Ras utilizes different effectors to mediate transformation and to deregulate cathepsin L expression and secretion in fibroblast and epithelial cells. PMID- 15352031 TI - Dissecting progressive stages of 5-fluorouracil resistance in vitro using RNA expression profiling. AB - Resistance to anticancer drugs such as the widely used antimetabolite 5 fluorouracil (FU) is one of the most important obstacles to cancer chemotherapy. Using GeneChip arrays, we compared the expression profile of different stages of FU resistance in colon cancer cells after in vitro selection of low-, intermediate- and high-resistance phenotypes. Drug resistance was associated with significant changes in expression of 330 genes, mainly during early or intermediate stage. Functional annotation revealed a majority of genes involved in signal transduction, cell adhesion and cytoskeleton with subsequent alterations in apoptotic response, cell cycle control, drug transport, fluoropyrimidine metabolism and DNA repair. A set of 33 genes distinguished all resistant subclones from sensitive progenitor cells. In the early stage, downregulation of collagens and keratins, together with upregulation of profilin 2 and ICAM-2, suggested cytoskeletal changes and cell adhesion remodeling. Interestingly, 6 members of the S100 calcium-binding protein family were suppressed. Acquisition of the intermediate-resistance phenotype included upregulation of the well-known drug resistance gene ABCC6 (ATP-binding cassette subfamily C member 6). The very small number of genes affected during transition to high resistance included the primary FU target thymidylate synthase. Although limited to an in vitro model, our data suggest that resistance to FU cannot be explained by known mechanisms alone and substantially involves a wide molecular repertoire. This study emphasizes the understanding of resistance as a time depending process: the cell is particularly challenged at the beginning of this process, while acquisition of the high-resistance phenotype seems to be less demanding. PMID- 15352032 TI - Expression of transforming growth factor-beta 1 and growth in soft agar differentiate prostate carcinoma-associated fibroblasts from normal prostate fibroblasts. AB - Carcinoma-associated fibroblasts (CAF) promote tumor progression of pre neoplastic epithelial cells. To investigate the basis of this phenomenon, we compared the properties of fibroblasts cultured from normal human prostate (NHPF) to prostate CAF. NHPF and CAF were assayed for growth potential, cell death and proliferative capacity by measuring population doubling time, cell cycle distribution and capability to form colonies in soft agar. Resistance to genotoxic (UV radiation: 0-50 J/cm2) and chemotoxic (0-200 nM Taxol) agents were compared between CAF and NHPF by measuring cell viability and cell cycle analysis. Transforming growth factor beta1 (TGF-beta1) immunoreactivity was assessed in non-malignant and malignant prostatic tissue. No detectable differences were found when comparing CAF and NHPF with respect to population doubling time, cell cycle distribution and response to genotoxic and chemotoxic agents. The mean number of colonies in soft agar was 120.5 for CAF vs. 18.2 for NHPF (p < 0.05). Because TGF-beta1 and matrix metalloproteinase (MMP)-9 have been associated with growth of fibroblasts in soft agar and tumor promotion, we measured the expression of these factors in NHPF and CAF by ELISA. There was no difference in expression of MMP-9; however, TGF-beta1 was expressed in higher concentrations in CAF than in NHPF (p < 0.0014). Furthermore, TGF-beta1 expression was higher in the carcinoma-associated stroma of prostate cancer tissue than stroma of non-malignant prostatic tissue. Increased capability of CAF as compared to NHPF to form colonies in soft agar may be due to a higher expression of TGF-beta1 and correlates with the ability of CAF to promote malignant progression of prostate epithelial cells. PMID- 15352033 TI - Fenretinide induces sustained-activation of JNK/p38 MAPK and apoptosis in a reactive oxygen species-dependent manner in neuroblastoma cells. AB - Fenretinide, which mediates apoptosis in neuroblastoma cells, is being considered as a novel therapeutic for neuroblastoma. The cytotoxic mechanisms of fenretinide, however, have not been fully elucidated. Sustained-activation of JNK and p38 MAPK signaling has been shown recently to have a pivotal role in stress induced apoptosis. Whether fenretinide activates the signaling in neuroblastoma cells is not known. In the present study, fenretinide induced sustained activation of both JNK and p38 MAPK in neuroblastoma cells. Pretreatment with the antioxidant L-ascorbic acid almost completely inhibited the accumulation of fenretinide-induced intracellular reactive oxygen species (ROS), activation of JNK and p38 MAPK and apoptosis. Intracellular ROS production and activation of stress signaling was not altered by fenretinide in resistant neuroblastoma cells. Our study demonstrates that in neuroblastoma cells, fenretinide induces sustained activation of JNK and p38 MAPK in an ROS-dependent manner and indicates that JNK and p38 MAPK signaling might mediate fenretinide-induced apoptosis. Our results also indicate that suppression of the fenretinide-induced ROS productive system and the downstream JNK and p38 MAPK signaling pathways causes neuroblastoma cells to become resistant to fenretinide. PMID- 15352034 TI - The tumor-suppressive reagent taurolidine is an inhibitor of protein biosynthesis. AB - Taurolidine has been successfully used as a disinfectant and to prevent the spreading and growth of tumor cells after surgical excision. However, the underlying mechanisms regarding its effects remain obscure. Here, we show that taurolidine treatment reduces endogenous levels of IkappaBalpha, p105, c-Jun, p53 and p27 in a dose-dependent manner in colon adenocarcinoma cells, which can be in part due to massive cell death. Because expression of tested proteins was affected by taurolidine, its influence on protein expression was studied. In the coupled transcription/translation system, taurolidine inhibited c-Jun expression with an IC50 value of 1.4 mM. There was no or little effect on transcription. In contrast, translation of c-Jun or p53 mRNA was completely inhibited by taurolidine. To determine which step of translation was affected, prominent complexes occurring in the course of translation were analyzed by density gradient centrifugation. In the presence of taurolidine, no preinitiation translation complex was assembled. Taurolidine also suppressed protein expression in bacteria. Based on our data, we conclude that taurolidine blocks a fundamental early phase of translation, which might explain its effects as a disinfectant and inhibitor of tumor growth. PMID- 15352035 TI - Induction of a metastatogenic tumor cell type by neurotransmitters and its pharmacological inhibition by established drugs. AB - The active migration of tumor cells, a crucial requirement for metastasis development and cancer progression, is regulated by signal substances including neurotransmitters. We investigated the migration of tumor cells within a three dimensional collagen matrix using time-lapse videomicroscopy and computer assisted analysis of the migration path. Tumor cell migration is induced by norepinephrine, dopamine and substance P. We show that this induced migration, using MDA-MB-468 breast and PC-3 prostate carcinoma cells, can be inhibited by using specific, clinically established receptor antagonists to the beta2 adrenoceptor, the D2 receptor, or the neurokinin-1 receptor, respectively. All of the investigated neurotransmitters significantly activated the cyclic adenosine monophosphate response element binding protein (CREB). Furthermore, microarray analysis revealed changes of gene expression toward a highly motile tumor cell type, including an upregulation of the alpha2 integrin, which is an essential adhesion receptor for collagen in migration. The gene for the tumor suppressor gelsolin was downregulated. These 2 critical alterations were confirmed on the protein level by flow-cytometry and immunoblotting, respectively. Neurotransmitters thus induce a metastatogenic tumor cell type by directly regulating gene expression and increased migratory activity, which can be prevented by established neurotransmitter antagonists. PMID- 15352036 TI - Identification of genes differentially expressed in human hepatocellular carcinoma by a modified suppression subtractive hybridization method. AB - To identify differentially expressed genes in human HCC in China, we applied a modified SSH method for cDNA subtraction. Such modification has made the method more effective for subtraction. We have obtained 36 and 24 differentially expressed cDNA fragments after modified SSH from 4 paired samples of human HCC and non-HCC tissues, respectively. Reverse Northern blotting analysis was performed to further identify the genes differentially expressed in the HCC and non-HCC tissue samples. There were 25 genes really overexpressed in HCC, and their corresponding encoding molecules may reflect the events of cell accelerated metabolism, proliferation, angiogenesis, anti-apoptosis, tumorigenesis (TLH107, TFH9) and the potential for metastasis. Of the 25 genes overexpressed in HCC, 5 were novel and their full-length cDNAs were cloned. These 5 novel genes are functionally associated with the occurrence and development of HCC according to the Database analysis. In the paired non-HCC tissues, there were 15 genes lowly or not expressed in HCC, and their encoding proteins function as tumor suppressors (TFA3, TFA11), acute-phase reactive proteins, and the blood plasma proteins that are mainly or exclusively synthesized in the liver. The distinct profiles of the differentially expressed genes in HCC and the paired non-HCC tissues have partially reflected the genetic alterations during HCC tumorigenesis. The novel HCC-specific gene TLH6 and the CT antigen encoding gene TLH107 may have diagnostic and therapeutic potentials in HCC and/or other solid cancers. PMID- 15352037 TI - Global gene expression profiles of human head and neck squamous carcinoma cell lines. AB - For genomewide monitoring and identification of biomarkers of head and neck squamous cell carcinoma (HNSCC), we have conducted a systematic characterization of gene expression profiles, using human cDNA microarrays containing 9K clones, in 25 HNSCC cell lines and 1 immortalized human oral keratinocyte cell line. We used normal human oral keratinocytes (NHOKs) as a reference. Our study showed that genes primarily involved in cell cycle regulation, oncogenesis, cell proliferation, differentiation, apoptosis and cell adhesion were widely altered in the 26 cell lines. Upregulated genes included known oncogenes, protein kinases, DNA-binding proteins and cell cycle regulators, while those commonly downregulated included differentiation markers, cell adhesion proteins, extracellular matrix proteins, structural proteins (keratins) and protease inhibitor proteins. Compared to NHOK, we observed a striking reduction in the expression of genes involved in terminal differentiation, suggesting that a loss in this process is an important signature of HNSCC. In addition, hierarchical clustering analysis as well as principal component analysis revealed 2 distinctive subtypes of gene expression patterns among the 26 cell lines, reflecting a degree of heterogeneity in HNSCC. By applying significance analysis of microarrays, 128 genes were selected for being distinctively expressed between the 2 groups. Genes differentially expressed in the 2 subgroups include cell proliferation-related genes, IGFBP6, EGFR and VEGFC; tumor suppression and apoptosis-related genes such as Tp53, Tp63; as well as cell cycle regulators such as CCND1 and CCND2 (cyclins D1 and D2), suggesting that the 2 subgroups might have undergone different pathways of carcinogenesis. PMID- 15352038 TI - Vegetable, fruit and meat consumption and potential risk modifying genes in relation to colorectal cancer. AB - Epidemiological evidence shows high red meat consumption to increase the risk of colorectal cancer, while the consumption of fruit and vegetables has been shown to be protective. Many genes have been identified that encode for enzymes involved in the metabolism of dietary carcinogens or anti-carcinogens. A study of 500 incident colorectal cancer cases and population controls, matched for age, sex and general practitioner, was conducted in the United Kingdom to investigate whether 6 such genes (CYP1A1, GSTT1, GSTM1, GSTP1, EPHX1 and NQO1) modify the relationship between diet and disease risk. Usual diet was estimated using a detailed questionnaire administered by interview. Fruit and vegetable consumption were both found to protect against colorectal cancer, while overall meat and red meat consumption were found to increase risk. There was some evidence of interaction between GSTT1 and vegetable consumption (p=0.006, not adjusted for multiple tests) but no evidence of interaction with GSTM1. The protective effect of vegetables was only seen in those with deficient or intermediate GSTT1 predicted phenotype [OR 0.3, 95% confidence interval (0.1, 0.6), and OR 0.6 (0.4, 0.96), OR 1.4 (0.3, 2.4) for those with fast phenotype], and a similar result was observed for cruciferous vegetables. There was also weak evidence of interaction between red meat intake and GSTT1 (p=0.06), GSTP1 (p=0.16, with p=0.02 after adjustment for potential confounders) and NQO1 predicted phenotype (p=0.01). Because of the multiple hypotheses tested in our study, these findings require independent confirmation. PMID- 15352039 TI - Expression of tumor necrosis factor receptor-associated factor 1 in nasopharyngeal carcinoma: possible upregulation by Epstein-Barr virus latent membrane protein 1. AB - EBV infection is associated with virtually all cases of undifferentiated NPC, and the EBV-encoded LMP1 is expressed in a proportion of cases. LMP1 has transforming functions similar to members of the TNF receptor family and activates intracellular signaling cascades through interaction with TRAFs. In B cells, expression of TRAF1 is in turn upregulated by LMP1. LMP1 signaling in epithelial cells may be affected by the presence or absence of TRAF1. By immunohistochemistry, we detected TRAF1 expression in 17 of 42 (40%) EBV+ undifferentiated NPCs. All 7 LMP1+ NPC biopsies were also TRAF1+. Using an RNAse protection assay, high-level TRAF1 expression was detected in an LMP1-expressing NPC-derived cell line (C15) and expression was weaker in 2 LMP1- cell lines (C17, C19). Finally, LMP1 upregulated TRAF1 expression in an EBV- keratinocyte cell line. Our results demonstrate that TRAF1 is expressed in NPC tumor cells in vivo and suggest that TRAF1 expression may be upregulated by LMP1 in NPC. An antiapoptotic function has been proposed for TRAF1, and this may be relevant for the pathogenesis of NPC. PMID- 15352040 TI - High-density methylation of p14ARF and p16INK4A in Epstein-Barr virus-associated gastric carcinoma. AB - Promoter hypermethylation of various tumor-related genes is extremely frequent in gastric carcinoma (GC) associated with Epstein-Barr virus (EBV). To investigate the significance of the promoter methylation in this type of GC, we examined the methylation densities of the promoter regions of p14ARF and p16INK4A in EBV associated (n=7) and EBV-negative (n=14) GC. Bisulfite sequencing demonstrated a high frequency of concurrent methylation of p14ARF and p16INK4A promoter regions in EBVaGC. Methylation was observed in all 29 CpG sites of p14ARF and all 16 sites of p16INK4A with equally high densities. In EBV-negative GC, the methylation profiles differed between the 2 genes. Promoter methylation was sporadic and variable in p14ARF, and only the last position of CpG in p14ARF was methylated at high frequency. High-density methylation in p16INK4A was observed in a subset of GC, but the first position of CpG was never methylated in EBV negative GC. These findings suggest the presence of mechanisms of de novo and maintenance methylation specific to EBVaGC that might be associated with EBV infection. PMID- 15352041 TI - RGD-modified anti-CD3 antibodies redirect cytolytic capacity of cytotoxic T lymphocytes toward alphavbeta3-expressing endothelial cells. AB - To redirect the lytic activity of cytotoxic T lymphocytes (CTL) toward tumor vascular endothelial cells, we prepared bifunctional proteins with specificity for both alphavbeta3 and CD3. Monocyclic RGD peptides (cRGDfK) were covalently coupled to an anti-CD3 monoclonal antibody at different peptide:protein ratios. The resulting RGDpep-anti-CD3 conjugates bound specifically to alphavbeta3 expressing endothelial cells. Displacement studies with radiolabeled alphavbeta3 ligand demonstrated that the alphavbeta3 binding affinity of RGDpep-anti-CD3 conjugates was elevated as compared to the non-conjugated RGDpep. IC50 values ranged from 150-1.1 nM, depending on the amount of coupled RGDpep molecules per antibody molecule. RGD modification did not affect the ability of anti-CD3 antibodies to bind to CTL. Furthermore, RGDpep-anti-CD3 was fully capable of activating T cells upon CD3 binding as was shown in a Jurkat/NFAT reporter-gene activation assay. All RGDpep-anti-CD3 conjugates were able to induce RGDpep, CD3 dependent lysis of human primary endothelial cells by anti-CD3/IL-2 activated human peripheral blood mononuclear cells (PBMC), with a significant induction of cytotoxicity observed at an E/T ratio as low as 10. Redirecting cytolytic activity reached up to 50% cytotoxicity using the conjugate with the highest RGD peptide load. Combining the good accessibility of tumor blood vessel endothelium for CTL with the efficiency of target cell killing warrants further investigations on anti-tumor effects of this type of conjugates in vivo. PMID- 15352042 TI - Investigation of MRP-1 protein and MDR-1 P-glycoprotein expression in invasive breast cancer: a prognostic study. AB - The efficacy of breast cancer treatment is limited by the development of resistance to various chemotherapeutic agents. We conducted a retrospective study of the expression of 2 drug resistance efflux pumps, MRP-1 and MDR-1 Pgp, in 177 invasive breast carcinomas. Immunohistochemical expression of these proteins was correlated with clinicopathologic characteristics as well as relapse-free survival (RFS) and overall survival (OS) times. MDR-1 Pgp was associated strongly with higher histologic grade (grade III). A highly significant association was shown between MDR-1 Pgp and MRP-1 expression (p < 0.01), 47.4% of patients expressing both proteins; MRP-1 was expressed in approximately 61% of patients and MDR-1, in approximately 66% of patients. No association was shown in the overall group between either MDR-1 Pgp or MRP-1 and any of the other clinicopathologic features. Kaplan-Meier analysis revealed that in a subset of patients with either high-grade (grade III) stage 1 (node-negative) or stage 2 (node-positive) tumours who were treated with surgery followed by adjuvant chemotherapy, MRP-1 expression in <25% of tumour cells at diagnosis was significantly associated with improved RFS (p < 0.02) and OS (p < 0.02). Using multivariate analysis, MRP-1 expression in <25% of tumour cells at diagnosis was identified as an independent, significant prognostic factor for RFS (p < 0.01) and OS (p < 0.01) in this patient group but not in other groups. In this subgroup, no significant correlation was observed between expression of MDR-1 Pgp and MRP-1. While the number of patients with high-grade tumours treated with adjuvant chemotherapy was small and further confirmatory research is warranted, it appears that assessment of MRP-1 expression at diagnosis may offer useful prognostic information in subgroups of patients with stage 1 or stage 2 high grade tumours who receive CMF-based adjuvant chemotherapy. Given the known substrate specificities of MRP-1, any mechanistic relationship between MRP-1 expression and CMF resistance remains unclear. No association was shown between MDR-1 Pgp expression and either RFS or OS time in any subgroup of patients. PMID- 15352043 TI - Carotenoids, vitamin A and risk of adenomatous polyp recurrence in the polyp prevention trial. AB - One trial reported beta-carotene supplementation was protective of adenomatous polyp recurrence in nonsmokers. We now examine the relation of serum and dietary carotenoids and vitamin A to adenomatous polyp recurrence in a subcohort of 834 participants in a low fat, high fiber, high fruit and vegetable dietary intervention, the Polyp Prevention Trial. Multivariate odds ratio (OR) and 95% confidence intervals (CI) of polyp recurrence were obtained using baseline or the average (first 3 years of the trial) carotenoid and vitamin A values after adjustment for covariates. Compared to the lowest quartile of baseline alpha carotene concentrations, the OR of multiple polyp recurrence for the highest quartile was 0.55 (95% CI = 0.30-0.99) and the OR of right-sided recurrence was 0.60 (95% CI = 0.37-0.95). Baseline dietary intakes of alpha-carotene and vitamin A from food with/without supplements were inversely associated with any recurrence (p for linear trend = 0.03-alpha-carotene; p = 0.004 and p = 0.007 intakes of vitamin A). Compared to the lowest quartile of averaged beta-carotene concentrations, the OR of multiple adenomas for the highest quartile was 0.40 (95% CI = 0.22-0.75) with an inverse trend (p = 0.02). The risk was inversely related to averaged: alpha-carotene concentrations and right-sided polyps; alpha carotene intake and recurrence of any, multiple and right-sided polyps; beta carotene intake and multiple adenoma recurrence; vitamin A from food (with supplements) and each adverse endpoint. Thus, alpha-carotene and vitamin A may protect against recurrence in nonsmokers and nondrinkers or be indicative of compliance or another healthy lifestyle factor that reduces risk. PMID- 15352044 TI - First pregnancy characteristics and subsequent breast cancer risk among young women. AB - There is growing evidence that perinatal factors associated with altered gestational hormones may influence subsequent breast cancer risk in the mother. Events occurring during the first pregnancy may be particularly important. In this matched case-control study, we investigated the relation between characteristics of a woman's first pregnancy and her later breast cancer risk using linked records from the New York State birth and tumor registries. Cases were 2,522 women aged 22 to 55 diagnosed with breast cancer between 1978 and 1995 and who had also completed a first pregnancy in New York State (NY) at least 1 year prior to diagnosis. Controls were 10,052 primiparous women not diagnosed with breast or endometrial cancer in NY and matched to cases on county of residence and date of delivery. Information on factors characterizing the woman's first pregnancy was obtained from the pregnancy record of each subject. The association of these factors to breast cancer risk was assessed using conditional logistic regression. Extreme prematurity (< 32 weeks gestational age) was associated with elevated maternal breast cancer risk [adjusted odds ratio (OR)=2.1, 95% confidence interval (CI) 1.2,3.9], as were abruptio placentae (OR = 1.8, CI 1.1,3.0) and multifetal gestation (OR=1.8, CI 1.1,3.0). Preeclampsia was associated with a marked reduction in breast cancer risk among women who bore their first child after age 30 (OR=0.3, CI 0.2,0.7) and in the first 3 years after delivery (OR=0.2 (0.1-0.9). These findings suggest that certain perinatal factors influence maternal breast cancer risk and offer indirect support for a role of gestational hormones, and particularly gestational estrogens, in the etiology of breast cancer. PMID- 15352045 TI - Endogenous sex hormones and subsequent breast cancer in premenopausal women. AB - Because of large intra-individual variation in hormone levels, few studies have investigated the relation of serum sex hormones to breast cancer (BC) in premenopausal women. We prospectively studied this relation, adjusting for timing of blood sampling within menstrual cycle. Premenopausal women (5,963), recruited to the Hormones and Diet in the Etiology of Breast Tumors (ORDET) cohort study, provided a blood sample in the 20-24th day of their menstrual cycle. After 5.2 years of follow-up, 65 histologically confirmed BC cases were identified and matched individually to 4 randomly selected controls. Sera, stored at -80 degrees C, were assayed blindly for dehydroepiandrosterone sulfate, total and free testosterone (FT), androstenedione, androstanediol-glucoronide, progesterone, 17 OH-progesterone, sex hormone-binding globulin, follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Fifty-five cases had information for multivariate analyses. Compared to controls, BC cases had shorter cycles and intervals between blood sampling and bleeding, and lower LH and FSH. FT was significantly associated with BC risk: relative risk (RR; adjusted for age, body mass index and ovarian cycle variables) of highest vs. lowest tertile was 2.85 [95% confidence interval (CI) = 1.11-7.33, p for trend = 0.030]. Progesterone was inversely associated with adjusted RR for highest vs. lowest tertile of 0.40 (95% CI = 0.15 1.08, p for trend = 0.077), significantly so in women with regular menses, where adjusted RR was 0.12 (95% CI = 0.03-0.52, p for trend = 0.005). These findings support the hypothesis that ovarian hyperandrogenism associated with luteal insufficiency increases the risk of BC in premenopausal women. PMID- 15352046 TI - Dietary soy and increased risk of bladder cancer: a prospective cohort study of men in Shanghai, China. AB - To verify our previous finding of a positive association between dietary soy and bladder cancer risk, we examined the association in a second, geographically distinct prospective cohort of Chinese subjects, the Shanghai Cohort Study. Briefly, 18,244 men aged 45-64 years were recruited between January 1986 and September 1989. As of December 31, 2002, 61 incident bladder cancer cases were identified. Information on soy consumption was obtained through in-person interviews at baseline using a food frequency questionnaire. Cox proportional hazard regression methods were used to estimate relative risks (RR) and their corresponding 95% confidence intervals (CI), with adjustment for age (years) at baseline interview, level of education and other potential confounders. Compared to men consuming soy less than once a week, the RR (95% CI) for those who consumed soy 1-<3 times per week, 3-<7 times a week and daily were 2.05 (0.80 5.29), 2.45 (0.89-6.76) and 4.61 (1.57-13.51), respectively (p for trend = 0.004), after adjustment for age, cigarette smoking and level of education. The soy-bladder cancer risk associations in smokers and non-smokers were comparable. The soy-bladder cancer relationship became stronger when the analysis was restricted to subjects with 2 or more years of follow-up. PMID- 15352047 TI - Smoking is associated with postmenopausal breast cancer in women with high levels of estrogens. AB - We investigated the association between smoking and risk of postmenopausal breast cancer in groups defined by high levels of estrogens, a factor known to enhance tumour progression. Two prospective cohorts of Swedish women provided 260 postmenopausal breast cancer cases and 514 controls. Blood samples were collected at baseline, and anthropometry, life-style factors and reproductive history had been assessed. Subjects were classified into quartiles with regard to the level of estrone, and into three categories with regard to estradiol. All analyses of the relation between smoking and breast cancer were repeated in different categories of these hormones. Logistic regression analysis, adjusted for matching factors, i.e., age at baseline, storage time and sub-cohort, yielded odds ratios (OR) with 95% confidence intervals (CI). Ever-smoking was associated with breast cancer in the top category of estrone, 2.02 (1.17-3.49). The highest risk was seen among ex-smokers, 2.96 (1.53-5.75). The pattern was similar for estradiol. Recent smoking cessation was associated with a high OR in top categories of estrone, 4.38 (1.27-15.2) and estradiol 10.0 (1.14-88.7). Smoking initiation before the age of 20 was associated with breast cancer in the top category of estrone, 2.73 (1.27-5.91). Several potential confounders were introduced into the statistical model, but none remained using backward selection. We conclude that ever-smoking was associated with the risk of breast cancer in women with high levels of estrone, and that ex-smoking was associated with breast cancer in women with high levels of estrone or estradiol. PMID- 15352048 TI - Anti-tumor activity of a combination of plasminogen activator and captopril in a human melanoma xenograft model. AB - Angiostatin, a proteolytic fragment of plasminogen consisting of the first 3 or 4 kringle domains, reduces tumor growth by specifically inhibiting tumor angiogenesis. Angiostatin is generated in vitro in a 2-step process. First, plasminogen is converted to plasmin by plasminogen activators. Next, plasmin excises the angiostatin fragment from plasminogen, a process requiring molecules that are able to donate a free sulfhydryl group. In this study, we investigated whether stimulation of in vivo angiostatin generation by administration of plasminogen activator and a free sulfhydryl group donor (FSD) has anti-tumor activity. First, we determined the optimal conditions for in vitro angiostatin generation by incubating murine plasma with different concentrations of plasminogen activator and/or the FSD captopril. Angiostatin generation was monitored by western blot analysis. Our results were extrapolated to the in vivo situation by administering the optimal dose of tissue-type plasminogen activator (tPA, i.v. injection 3 times/week) and captopril (in drinking water) to mice and analyzing the presence of angiostatin in the circulation. Angiostatin was readily detectable in mice receiving both tPA and captopril, but not in mice receiving either one of the agents. Finally, the anti-tumor activity of the tPA/captopril treatment was tested in a human melanoma xenograft model. Administration of tPA alone had only a marginal effect on tumor growth. Captopril alone reduced tumor growth by about 60%, whereas treatment with both captopril and tPA resulted in 83% inhibition of tumor growth. PMID- 15352049 TI - Efficacy of transferrin-conjugated paclitaxel-loaded nanoparticles in a murine model of prostate cancer. AB - Chemotherapy remains the preferred choice of treatment for prostate cancer but modest drug response and significant toxicity by conventional methods of administration limit their efficacy. In our study, we determined the efficacy of paclitaxel (Tx)-loaded biodegradable nanoparticles (NPs) on tumor inhibition. We hypothesized that NPs following conjugation to transferrin (Tf) ligand (NPs-Tf) would enhance the therapeutic efficacy of the encapsulated drug. The antiproliferative activity of NPs was determined in human prostate cancer cell line (PC3) and their effect on tumor inhibition in a murine model of prostate cancer. NPs (approximately 220 nm in diameter, 5.4% w/w drug loading) under in vitro conditions exhibited sustained release of the encapsulated drug (60% release in 60 days). The IC50 (concentration of drug for 50% inhibition of cell growth) of the drug with Tf-conjugated NPs (Tx-NPs-Tf) was about 5-fold lower than that with unconjugated NPs (Tx-NPs) or drug in solution. Animals that received a single-dose intratumoral injection of Tx-NPs-Tf (Tx dose= 4 mg/kg) demonstrated complete tumor regression and greater survival rate than those that received either Tx-NPs or Tx-Cremophor EL formulation. In conclusion, sustained release NPs demonstrated greater antitumor activity following their conjugation to Tf ligand. PMID- 15352050 TI - Accuracy of human papillomavirus testing in primary screening of cervical neoplasia: results from a multicenter study in India. AB - The knowledge that cervical neoplasia are caused by human papillomavirus (HPV) infection has led to the evaluation of its role in screening. We evaluated the accuracy of HPV testing by Hybrid capture II (HC II) method in detecting cervical intraepithelial neoplasia grade 2 and 3 (CIN 2 and 3) lesions in 4 cross sectional studies with common protocol and questionnaire in 3 different locations (Kolkata, Mumbai and Trivandrum) in India. These studies involved 18,085 women aged 25-65 years. The reference standard for final diagnosis was a combination of colposcopy/biopsy. All women were investigated with colposcopy and 3,116 received directed biopsy. The sensitivity of HPV testing for detecting CIN 2-3 lesions varied from 45.7% to 80.9% across the study sites; the specificity varied from 91.7% to 94.6% and the positive predictive value from 6.7% to 13.7%. Retesting of 298 randomly chosen denatured samples in France revealed an agreement rate of 85.9% and a kappa-value of 0.72. Although HPV testing seems to be a promising approach for cervical cancer prevention, a large range in sensitivity was observed in our study, possibly due to variations in the quality of specimen collection and reference standards. A higher sensitivity was associated with the center performing the test well. Further developments in terms of more reproducible, less expensive and less sophisticated testing are essential to make the test feasible and effective in low-resource settings. PMID- 15352051 TI - Childhood body mass index and later cancer risk: a 50-year follow-up of the Boyd Orr study. AB - Associations between childhood BMI and adult cancer risk were investigated in a historical cohort study based on the Carnegie ("Boyd Orr") Survey of Diet and Health in Pre-War Britain (1937-9). In 14 centres in England and Scotland, children had their height and weight measured. We included 2,347 individuals aged between 2 and 14 years 9 months at the time of measurement, who were traced through the National Health Service Central Register. Relative cancer risk (registration or death) was estimated in relation to age- and sex-specific BMI SD scores. We studied associations with (i) all cancers, (ii) cancer groups stratified according to their relationship to smoking and (iii) certain site specific cancers. In the 50 years of follow-up, 188 men and 192 women developed cancer. There was a 9% increase (95% CI -3 to 22%) in risk of cancer in adulthood per SD increase in BMI measured in childhood. There was no evidence of confounding by childhood or adulthood socioeconomic position, other anthropometric variables, childhood energy intake or birth order. There was a 30% increase (95% CI 10-54%) in risk of smoking-related cancers per SD increase in childhood BMI. There was no relationship between BMI and cancers not related to smoking. Associations for all cancers and non-smoking-related cancers tended to be stronger in children who were measured at an older (>8 years) rather than a younger (< or =8 years) age. We conclude that childhood BMI is related to increased risk of cancer in later life, particularly smoking-related cancers. PMID- 15352052 TI - Low levels of mycophenolic acid induce differentiation of human neuroblastoma cell lines. PMID- 15352054 TI - Reduced protein adsorption at solid interfaces by sugar excipients. AB - Sugar excipients are shown to reduce the adsorption of ribonuclease A, bovine serum albumin, and hen egg white lysozyme at the liquid-solid interface. The amount of protein adsorbed decreased as the concentration of the sugar increased. At the same sugar concentration, the ability of sugars to reduce protein adsorption followed the trend: trisaccharides > disaccharides > 6-carbon polyols > monosaccharides. This trend in adsorbed protein amounts among sugars was explained by stabilization of the protein native state in solution by the sugar excipients. The heat of solution of the amorphous saccharide was found to correlate with the amount of protein adsorbed. PMID- 15352055 TI - Biooxidation of pyrite by defined mixed cultures of moderately thermophilic acidophiles in pH-controlled bioreactors: significance of microbial interactions. AB - The oxidative dissolution of pyrite (FeS2) by pure and mixed cultures of moderately thermophilic acidophiles was studied in shake flask cultures and in pH controlled bioreactors, incubated at 45 degrees C. Various combinations of seven eubacteria (a Leptospirillum sp. (MT6), Acidimicrobium ferrooxidans, Acidithiobacillus caldus, an Alicyclobacillus sp. (Y004), and three Sulfobacillus spp.) and one archaeon (Ferroplasma sp. MT17) were examined. Pyrite dissolution was determined by measuring changes in soluble iron and generation of acidity, and microbial populations were monitored using a combined culture-dependent (plate counts) and culture-independent (fluorescent in situ hybridization) approach. In pure cultures, the most efficient pyrite-oxidizing acidophile was Leptospirillum MT6, which was unique among the prokaryotes used in being obligately autotrophic. Mixed cultures of Leptospirillum MT6 and the sulfur oxidizer At. caldus generated more acidity than pure cultures of the iron oxidizer, though this did not necessarily enhance pyrite dissolution. In contrast, a mixed culture of Leptospirillum MT6 and the obligate heterotroph Alicyclobacillus Y004 oxidized pyrite more rapidly and more completely than a pure culture of Leptospirillum MT6, in synchronized bioreactors. Although the autotroph, At. caldus, and the "heterotrophically inclined" iron-oxidizer, Am. ferrooxidans, were both ineffective at leaching pyrite in pure culture, a mixed culture of the two bacteria was able to accelerate dissolution of the mineral. Concentrations of dissolved organic carbon accumulated to >100 mg/L in some mixed cultures, and the most effective bioleaching systems were found to be consortia containing both autotrophic and heterotrophic moderate thermophiles. A mixed culture comprising the autotrophs Leptospirillum MT6 and At. caldus, and the heterotroph Ferroplasma MT17, was the most efficient of all of those examined. Mutualistic interactions between physiologically distinct moderately thermophilic acidophiles, involving transfer of organic and inorganic carbon and transformations of iron and sulfur, were considered to have critical roles in optimizing pyrite dissolution. PMID- 15352056 TI - Estimating the potential refolding yield of recombinant proteins expressed as inclusion bodies. AB - Recombinant protein production in bacteria is efficient except that insoluble inclusion bodies form when some gene sequences are expressed. Such proteins must undergo renaturation, which is an inefficient process due to protein aggregation on dilution from concentrated denaturant. In this study, the protein-protein interactions of eight distinct inclusion-body proteins are quantified, in different solution conditions, by measurement of protein second virial coefficients (SVCs). Protein solubility is shown to decrease as the SVC is reduced (i.e., as protein interactions become more attractive). Plots of SVC versus denaturant concentration demonstrate two clear groupings of proteins: a more aggregative group and a group having higher SVC and better solubility. A correlation of the measured SVC with protein molecular weight and hydropathicity, that is able to predict which group each of the eight proteins falls into, is presented. The inclusion of additives known to inhibit aggregation during renaturation improves solubility and increases the SVC of both protein groups. Furthermore, an estimate of maximum refolding yield (or solubility) using high performance liquid chromatography was obtained for each protein tested, under different environmental conditions, enabling a relationship between "yield" and SVC to be demonstrated. Combined, the results enable an approximate estimation of the maximum refolding yield that is attainable for each of the eight proteins examined, under a selected chemical environment. Although the correlations must be tested with a far larger set of protein sequences, this work represents a significant move beyond empirical approaches for optimizing renaturation conditions. The approach moves toward the ideal of predicting maximum refolding yield using simple bioinformatic metrics that can be estimated from the gene sequence. Such a capability could potentially "screen," in silico, those sequences suitable for expression in bacteria from those that must be expressed in more complex hosts. PMID- 15352057 TI - Real-time update of calibration model for better monitoring of batch processes using spectroscopy. AB - In order to reduce the large calibration matrix usually required for calibrating multiwavelength optical sensors, a simple algorithm based on the addition in process of new standards is proposed. A small calibration model, based on 14 standards, is periodically updated by spectra collected on-line during fermentation operation. Concentrations related to these spectra are reconciled into best-estimated values, by considering carbon and oxygen balances. Using this method, fructose, acetate, and gluconacetan were monitored during batch fermentations of Gluconacetobacter xylinus 12281 using mid-infrared spectroscopy. It is shown that this algorithm compensates for noncalibrated events such as production or consumption of by-products. The standard error of prediction (SEP) values were 0.99, 0.10, and 0.90 g/L for fructose, acetate, and gluconacetan, respectively. By contrast, without an updating of the calibration model, the SEP values were 2.46, 0.92, and 1.04 g/L for fructose, acetate, and gluconacetan, respectively. Using only 14 standards, it was therefore possible to approach the performance of an 88-standard-based calibration model having SEP values of 1.11, 0.37, and 0.79 g/L for fructose, acetate, and gluconacetan, respectively. Therefore, the proposed algorithm is a valuable approach to reduce the calibration time of multiwavelength optical sensors. PMID- 15352058 TI - Change of extracellular cAMP concentration is a sensitive reporter for bacterial fitness in high-cell-density cultures of Escherichia coli. AB - Guanosine-3',5'-tetraphosphate (ppGpp) and sigmaS, two regulators of the starvation response of Escherichia coli, have received increasing attention for monitoring cell physiological changes in production processes, although both are difficult to quantify. The kinetics of cAMP formation and degradation were not yet investigated in such processes, although the complex regulation of cAMP by synthesis, release, and degradation in connection with straightforward methods for analysis renders it a highly informative target. Therefore, we followed the cAMP concentration in various nonrecombinant and in four different recombinant glucose-limited fed-batch processes in different production scales. The intracellular cAMP concentration increases strongly at the end of the batch phase. Most cAMP is released to the cultivation medium. The rates of accumulation and degradation of extracellular cAMP are growth-rate-dependent and show a distinct maximum at a growth rate of about 0.35 h(-1). At very low growth rates, below 0.05 h(-1), extracellular cAMP is not produced but rather degraded, independent of whether this low growth rate is caused by glucose limitation or by the high metabolic load of recombinant protein production. In contrast to intracellular cAMP, which is highly unstable, analysis of extracellular cAMP is simpler and the kinetics of accumulation and degradation reflect well the physiological situation, including unlimited growth, limitation, and severe starvation of a production host. PMID- 15352059 TI - Establishment of FUT8 knockout Chinese hamster ovary cells: an ideal host cell line for producing completely defucosylated antibodies with enhanced antibody dependent cellular cytotoxicity. AB - To generate industrially applicable new host cell lines for antibody production with optimizing antibody-dependent cellular cytotoxicity (ADCC) we disrupted both FUT8 alleles in a Chinese hamster ovary (CHO)/DG44 cell line by sequential homologous recombination. FUT8 encodes an alpha-1,6-fucosyltransferase that catalyzes the transfer of fucose from GDP-fucose to N-acetylglucosamine (GlcNAc) in an alpha-1,6 linkage. FUT8(-/-) cell lines have morphology and growth kinetics similar to those of the parent, and produce completely defucosylated recombinant antibodies. FUT8(-/-)-produced chimeric anti-CD20 IgG1 shows the same level of antigen-binding activity and complement-dependent cytotoxicity (CDC) as the FUT8(+/+)-produced, comparable antibody, Rituxan. In contrast, FUT8(-/-)-produced anti-CD20 IgG1 strongly binds to human Fcgamma-receptor IIIa (FcgammaRIIIa) and dramatically enhances ADCC to approximately 100-fold that of Rituxan. Our results demonstrate that FUT8(-/-) cells are ideal host cell lines to stably produce completely defucosylated high-ADCC antibodies with fixed quality and efficacy for therapeutic use. PMID- 15352060 TI - Arabidopsis glycosyltransferases as biocatalysts in fermentation for regioselective synthesis of diverse quercetin glucosides. AB - Regioselectivity of glycosyltransferases offers an important means to overcome the limitations of chemical synthesis of small molecule glycosides. In this study we explore a large multigene family of UDP-glucose:glycosyltransferases of Arabidopsis for their potential as novel biocatalysts for in vitro synthesis and whole-cell catalysis. We used quercetin as a substrate for this study because the flavonol and its glycosides have important medicinal properties and the metabolite provides a complex structure for regioselective glucosylation. We analyzed the activity of 91 recombinant enzymes for in vitro activity toward quercetin and discovered 29 that are capable of glucosylating the substrate. We demonstrate the first enzymic synthesis of a range of glucosides in vitro, including the 3-O-, 7-O-, 3'-O-, and 4'-O-monoglucosides, 3,7-di-O-glucoside, and 7,3'-di-O-glucoside. We also show that the regioselectivity of glucosylation can be maintained when the enzymes are used as whole-cell biocatalysts in Escherichia coli. PMID- 15352061 TI - Williopsis californica, Williopsis saturnus, and Pachysolen tannophilus: novel microorganisms for stereoselective oxidation of secondary alcohols. AB - A screening of 416 microorganisms from different taxonomical groups (bacteria, actinomycetes, yeasts, and filamentous fungi) has been performed looking for active strains in the stereoselective oxidation of secondary alcohols. The working collection was composed of 71 bacterial strains, 45 actinomycetes, 59 yeasts, 60 basidiomycetes, 33 marine fungi, and 148 filamentous fungi. All microorganisms selected were mesophilic. Yeasts were the most active microbial group in the whole-cell-catalyzed oxidation. Williopsis californica, Williopsis saturnus, and Pachysolen tannophilus were the strains of greatest interest, both as growing cells and as resting cells. The oxidation of the alcohols takes place when cells are in the stationary growth phase (after 48 h of culture). These three strains are S-stereoselective for the oxidation of racemic secondary alkanols and show stereospecificity in the oxidation of menthol or neo-menthol, whereas iso-menthol is not oxidized. In the case of the 1-tetrahydronaphtol enantiomers, only the S-enantiomer is oxidized. The three strains were immobilized by entrapment using agarose and agar from algae of the Gracilaria genus. The agarose derivatives displayed significant improvement in the stereospecificity of the reactions. PMID- 15352062 TI - Extreme scale-down of expanded bed adsorption: Purification of an antibody fragment directly from recombinant E. coli culture. AB - Scale-down is a methodology that combines the use of very small volumes of process fluid in dedicated devices to predict accurately the behaviour of process scale biotechnological unit operations and for the production of comparable material for use in further devices which, taken together, facilitate the mimic of a complete full-scale process. This article provides the rationale behind the development of a small-scale mimic and demonstrates the use of a highly scaled down expanded bed to predict hydrodynamic, kinetic, and adsorptive performance using less than 5-mL sample volumes. Data acquired on a specially developed 1.9 mm ID column was compared with that obtained in a standard 25 mm ID column. A homogenised E. coli system expressing an antibody fragment (F(ab)) adsorbed onto an rProtein A matrix was used to characterise the full adsorptive performance. Breakthrough curve studies using BSA in buffer were used to characterise binding kinetics. Performance at the two scales was comparable both in terms of expansion, axial dispersion, binding isotherms, and elution behaviour of the antibody fragment. The eluted F(ab) material was further purified by ion exchange chromatography to demonstrate the similarity between the profile of the product material obtained at both scales. The high level of scale-down (approximately 200 fold) provides for rapid process evaluation early in development, where material is at a premium and where a fast appreciation of the likely merits of one process strategy will lead to greater confidence in process selection and more robust flowsheets. PMID- 15352063 TI - Anaerobic hydrogen production with an efficient carrier-induced granular sludge bed bioreactor. AB - A novel bioreactor containing self-flocculated anaerobic granular sludge was developed for high-performance hydrogen production from sucrose-based synthetic wastewater. The reactor achieved an optimal volumetric hydrogen production rate of approximately 7.3 L/h/L (7,150 mmol/d/L) and a maximal hydrogen yield of 3.03 mol H2/mol sucrose when it was operated at a hydraulic retention time (HRT) of 0.5 h with an influent sucrose concentration of 20 g COD/L. The gas-phase hydrogen content and substrate conversion also exceeded 40 and 90%, respectively, under optimal conditions. Packing of a small quantity of carrier matrices on the bottom of the upflow reactor significantly stimulated sludge granulation that can be accomplished within 100 h. Among the four carriers examined, spherical activated carbon was the most effective inducer for granular sludge formation. The carrier-induced granular sludge bed (CIGSB) bioreactor was started up with a low HRT of 4-8 h (corresponding to an organic loading rate of 2.5-5 g COD/h/L) and enabled stable operations at an extremely low HRT (up to 0.5 h) without washout of biomass. The granular sludge was rapidly formed in CIGSB supported with activated carbon and reached a maximal concentration of 26 g/L at HRT = 0.5 h. The ability to maintain high biomass concentration at low HRT (i.e., high organic loading rate) highlights the key factor for the remarkable hydrogen production efficiency of the CIGSB processes. PMID- 15352064 TI - Adsorption of fluorescein by protein crystals. AB - Adsorption characteristics of native and cross-linked lysozyme crystals were examined using fluorescein as model adsorbate. The adsorption isotherms exhibited Langmuir or linear behavior. The affinity constant (b1) and the adsorption capacity (Qsat) for fluorescein were found to depend on the type and concentration of co-solute present in the solution. The dynamics of adsorption isotherm transition from Langmuir to linear showed that affinity of lysozyme for solutes increases in the order 2-(cyclohexylamino)ethanesulphonic acid (CHES), 4 morpholinepropanesulphonic acid (MOPS), acetate, fluorescein. Furthermore, the crystal morphology, the degree of cross-linking of the crystals, and, in particular, solution pH were identified as factors determining fluorescein adsorption by the lysozyme crystals. These factors seem to affect crystal capacity for the solute more than affinity for the solute. Adsorption of fluorescein by cross-linked tetragonal lysozyme crystals was exponentially dependent on the lysozyme net charge calculated from the final solution pH. The 3 5-fold increase in the fluorescein adsorption as a result of cross-linking is presumably due to the increasing hydrophobicity of the lysozyme crystal. PMID- 15352065 TI - Neurotransmitter analog tethered to a silicon platform for neuro-BioMEMS applications. AB - The design of chemically well-defined, machinable surfaces containing neuroactive molecules offers potential for fundamental neuroscience and clinical neural engineering applications. Here we report the assembly and characterization of silicon platforms containing a tethered form of muscimol. Muscimol, an analog of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), is a potent agonist at postsynaptic GABA(A) and GABA(C) receptors. Surfaces were assembled using covalent avidin conjugation to silanized silicon followed by high-affinity avidin-biotin binding of a biotinylated derivative of muscimol (muscimol-biotin). Contact angle measurements, ellipsometry, and X-ray photoelectron spectroscopy (XPS) were conducted to characterize the wettability, thickness, and chemical composition of progressively deposited surface layers. The data demonstrate successful incorporation of a neurotransmitter analog as part of a layered, silicon-based structure possessing robust and specific biomolecular composition. These findings represent a step toward the design of platforms for applications involving control and modulation of neural signaling. PMID- 15352066 TI - Precipitation by polycation as capture step in purification of plasmid DNA from a clarified lysate. AB - The demand for highly purified plasmids in gene therapy and plasmid-based vaccines requires large-scale production of pharmaceutical-grade plasmid. Large scale purification of plasmid DNA from bacterial cell culture normally includes one or several chromatographic steps. Prechromatographic steps include precipitation with solvents, salts, and polymers combined with enzymatic degradation of nucleic acids. No method alone has so far been able to selectively capture plasmid DNA directly from a clarified alkaline lysate. We present a method for selective precipitation of plasmid DNA from a clarified alkaline lysate using polycation poly(N, N'-dimethyldiallylammonium) chloride (PDMDAAC). The specific interaction between the polycation and the plasmid DNA resulted in the formation of a stoichiometric insoluble complex. Efficient removal of contaminants such as RNA, by far the major contaminant in a clarified lysate, and proteins as well as 20-fold plasmid concentration has been obtained with about 80% recovery. The method utilizes a inexpensive, commercially available polymer and thus provides a capture step suitable for large-scale production. PMID- 15352068 TI - Lentiviral vectors for treating and modeling human CNS disorders. AB - Vectors based on lentiviruses efficiently deliver genes into many different types of primary neurons from a broad range of species including man and the resulting gene expression is long term. These vectors are opening up new approaches for the treatment of neurological diseases such as Parkinson's disease (PD), Huntington's disease (HD), and motor neuron diseases (MNDs). Numerous animal studies have now been undertaken with these vectors and correction of disease models has been obtained. Lentiviral vectors also provide a new strategy for in vivo modeling of human diseases; for example, the lentiviral-mediated overexpression of mutated human alpha-synuclein or huntingtin genes in basal ganglia induces neuronal pathology in animals resembling PD and HD in man. These vectors have been refined to a very high level and can be produced safely for the clinic. This review will describe the general features of lentiviral vectors with particular emphasis on vectors derived from the non-primate lentivirus, equine infectious anemia virus (EIAV). It will then describe some key examples of genetic correction and generation of genetic animal models of neurological diseases. The prospects for clinical application of lentiviral vectors for the treatment of PD and MNDs will also be outlined. PMID- 15352069 TI - Safe two-plasmid production for the first clinical lentivirus vector that achieves >99% transduction in primary cells using a one-step protocol. AB - We report the design of a unique two-plasmid production system for the first lentiviral vector to be evaluated in humans, VRX496. VRX496 is an optimized VSV-G pseudotyped vector derived from HIV-1 that expresses antisense to the HIV envelope gene. We found that a two-plasmid approach to production resulted in higher vector production titers when compared with a three-plasmid approach, which is particularly important for vector production at the large scale. Therefore, we carefully designed a single packaging construct, VIRPAC, for safety by reducing its homology with VRX496 and by insertion of functionally validated genetic elements designed to reduce the risk of generation of a replication competent lentivirus (RCL). A native cis-acting ribozyme is used to prevent read through into the envelope gene from the upstream gag-pol genes in the packaging vector, thus preventing RNAs containing gag-pol and env together for comparable safety to a three-plasmid system. We demonstrate that there is no significant in vivo vector mobilization using a primary SCID-hu mouse transplantation model, which correlates with the presence of an anti-HIV payload and suggests that inclusion of antisense may be a useful tool to restrict mobilization in other vector constructs. Gene transfer is achieved using a one-step transduction procedure that is simple and clinically translatable, which reaches stable transduction efficiencies of >99% in CD4+ T lymphocytes within 3 days of culture initiation. PMID- 15352070 TI - Hepatocyte-specific gene expression from integrated lentiviral vectors. AB - BACKGROUND: For many applications, efficient gene therapy will require long-term, organ-specific therapeutic gene expression. Lentiviral vectors based on HIV-1 are promising gene delivery vehicles due to their ability to integrate transgenes into non-dividing cells. Many experimental vectors express transgenes under the control of the cytomegalovirus (CMV) immediate-early gene promoter. Although this promoter directs strong gene expression in vitro, it may be shut off rapidly in vivo. This study explores the potential of HIV-1-based vectors to transduce hepatocytes and compares gene expression from different promoters in integrated vectors. METHODS: HIV-1-based vector plasmids expressing the green fluorescent protein (GFP) under the control of the CMV promoter, the alpha-1 antitrypsin gene promoter or promoters derived from the hepatitis B virus (HBV) genome were used to compare expression in transfected and transduced cell lines. RESULTS: Hepatocyte cell lines differed strikingly in their transfectability. Transduction with replication-deficient HIV-1-based vector particles incorporating the different promoter elements was uniformly effective in hepatocyte and non hepatocyte lines. However, in hepatocytes, only the CMV, alpha-1 antitrypsin and HBV core but not HBV surface promoters were able to produce GFP expression. Addition of the HBV enhancer 2 element improved the transducing ability of the HBV surface promoter and suppressed expression in non-hepatocytes increasing specificity for hepatocytes. CONCLUSIONS: Integrated lentiviral vectors can be used to direct transgene expression in liver cells both promiscuously and specifically. Promoters derived from the alpha-1 antitrypsin gene or HBV are alternatives to the CMV promoter. Inclusion of the HBV enhancer 2 permits strong liver-specific gene expression in vitro. PMID- 15352071 TI - Structural and functional healing of critical-size segmental bone defects by transduced muscle-derived cells expressing BMP4. AB - BACKGROUND: Our previous studies have shown that muscle-derived cells, including a population of muscle stem cells, transduced with a retroviral vector expressing bone morphogenetic protein 4 (BMP4) can improve the healing of critical-size calvarial defects. However, we did not evaluate the functionality of the healed bone. The purpose of this study was to determine whether primary muscle-derived cells transduced with retroBMP4 can heal a long bone defect both structurally and functionally. METHODS: Primary muscle-derived cells were genetically engineered to express BMP4 and were implanted into 7-mm femoral defects created in syngeneic rats. Muscle-derived cells transduced with retroLacZ were used in the control group. Bone healing was monitored by radiography, histology, and biomechanical testing at designated time points. RESULTS: Most of the defects treated with muscle-derived cells expressing BMP4 formed bridging callous by 6 weeks after surgery, and exhibited radiographically evident union at 12 weeks after cell implantation. Histological analysis at 12 weeks revealed that the medullary canal of the femur was restored and the cortex was remodeled between the proximal and distal ends of each BMP4-treated defect. In contrast, the defects treated with muscle-derived cells expressing beta-galactosidase displayed nonunion at all tested time points. An evaluation of the maximum torque-to-failure in the treatment group indicated that the healed bones possessed 77 +/- 28% of the strength of the contralateral intact femora. Torsional stiffness and energy-to failure were not significantly different between the treated and intact limbs. CONCLUSIONS: This study demonstrated that primary muscle-derived cells transduced with retroBMP4 can elicit both structural and functional healing of critical-size segmental long bone defects created in rats. PMID- 15352072 TI - Marked inhibition of retinal neovascularization in rats following soluble-flt-1 gene transfer. AB - BACKGROUND: In mouse models of retinopathy of prematurity (ROP) inhibitors of vascular endothelial growth factor (VEGF) functions administered systemically completely block retinal neovascularization. In contrast, selective ocular VEGF depletion has achieved an approx. 50% inhibition of retinal neovascular growth. It is unclear whether a more complete inhibition of new blood vessel development can be obtained with an anti-VEGF therapy localized to the eye. Therefore, the objective of the present study was to determine the effect of local anti-VEGF therapy in a different animal model which closely mimics human ROP. METHODS: Rats were exposed to alternating cycles of high and low levels of oxygen for 14 days immediately after birth; thereafter, they were intravitreally injected with an adenoviral vector expressing a secreted form of the VEGF receptor flt-1 (Ad.sflt), which acts by sequestering VEGF. Contralateral eyes were injected with the control vector carrying the reporter gene expressing beta-galactosidase (Ad.betaGal). RESULTS: At the peak of retinal neovascular growth, i.e. post-natal day 21 (P21), we observed up to 97.5% decrease in retinal neovascularization in animals injected with Ad.sflt. At the end of observation (P28), no significant difference in retinal vessel number was detected in both oxygen-injured and normoxic Ad.sflt-treated retinas compared with untreated or Ad.betaGal-treated retinas. CONCLUSION: Adenoviral-mediated sflt-1 gene transfer induces a near complete inhibition of ischemia-induced retinal neovascularization in rats without affecting pre-existing retinal vessels. PMID- 15352073 TI - Differential rAAV2 transduction efficiencies and insulin secretion profiles in pure and co-culture models of human enteroendocrine L-cells and enterocytes. AB - BACKGROUND: Cell-based therapies for treating insulin-dependent diabetes (IDD) can provide a more physiologic regulation of blood glucose levels in a less invasive fashion than insulin injections. Previously, we developed an engineered human enteroendocrine L-cell model for regulated insulin release via recombinant adeno-associated virus serotype 2, or rAAV2, transduction. The aim of this study was to evaluate the efficiency and selectivity of rAAV2-mediated insulin gene delivery to enteroendocrine L-cells in co-culture with a prevailing number of enterocytes, which are the predominant cell type in intestinal epithelium. METHODS: We tested rAAV2 transduction in pure and co-culture models of human cell lines of enterocytes (Caco-2 and T84 cell lines) and enteroendocrine L-cells (NCI H716 cell line). Non-viral, chemical-mediated transfection was used as a control. Transduced and transfected co-cultures were subjected to insulin secretion studies. RESULTS: In pure cultures, rAAV2 exhibited a low transduction efficiency towards both Caco-2 and T84 enterocytes, as opposed to a strong reporter expression in permissive NCI-H716 L-cells. In co-cultures of NCI-H716 L-cells and Caco-2 or T84 enterocytes, rAAV2 exhibited differential transduction efficiency with a strong preference towards NCI-H716 L-cells. The rAAV2-transduced co culture achieved regulated insulin release against stimulation, whereas the chemically transfected co-culture failed to respond. CONCLUSIONS: This study demonstrated that rAAV2-mediated insulin gene transfer can differentiate human intestinal cell types in vitro, in particular enterocyte and enteroendocrine L cell lines. We consider the AAV2 vector a useful tool in developing enteroendocrine L-cell-specific insulin gene delivery for IDD treatment, in terms of AAV2 avoiding enterocytes and targeting selectively L-cells. PMID- 15352074 TI - Vesicular stomatitis virus glycoprotein: a transducing coat for SFV-based RNA vectors. AB - BACKGROUND: Semliki Forest virus (SFV) vectors have a great potential for the induction of protective immunity in a large number of clinical conditions including cancer. Such a potential accounts for the huge efforts made to improve the in vivo expression from SFV vectors. It is noteworthy that efficient in vivo expression strongly relies on the ability to deliver high-titre vectors. To achieve this, the generation of recombinant SFV particles, using independent expression systems for structural SFV genes, has been proposed. However, despite several modifications in the production process, a risk of contamination with replication-competent, or partially recombined, virus has remained. METHODS: Here, we exploit the ability of the vesicular stomatitis virus glycoprotein (VSV G), expressed in trans, to hijack full-length genomic SFV RNA into secreted virus like particles (VLPs). To allow SFV vector mobilisation, we designed a CMV driven SFV vector in which the internal 26S promoter has been extensively mutated. With this vector, mobilisation events were monitored using the Green Fluorescent Protein (GFP). The production procedure involves a sequential transfection protocol, of plasmids expressing the VSV-G and the SFV vector respectively. RESULTS: We show that the VLPs are effective for cellular delivery of SFV vectors in a broad range of human and non-human cellular targets. Furthermore, production of VLPs is easy and allows, through concentration, the harvest of high-titre vector. CONCLUSIONS: The present paper describes a convenient process aimed at mobilising full length SFV vectors. A major issue to consider, while developing clinically relevant gene transfer vectors, is the risk of undesirable generation of replication competent by-products. Importantly, as the VSV-G gene shares no homology with the SFV genome, our VLPs offer a strong guarantee of biosafety. PMID- 15352075 TI - Improved non-viral transfection of glial and adult neural stem cell lines and of primary astrocytes by combining agents with complementary modes of action. AB - BACKGROUND: Rational design of gene vectors for therapeutic applications requires understanding of transfection mechanisms. In this study, multiple transfection assays revealed complementary mechanisms between two commonly used transfection agents. This finding was then exploited to produce improved transfection outcomes. METHODS AND RESULTS: Rat C6 glial cells, adult rat hippocampal progenitor cells and primary astrocytes were transfected using Lipofectamine (LA) or polyethylenimine (PEI), in vitro. Although LA- and PEI-transfected populations expressed the same total level of transgene product, LA transfected considerably more cells than PEI (approximately 20 vs. 14%). A fluorescently labelled plasmid and time-course analysis, involving both flow cytometry and confocal microscopy, were used to explain this apparent discrepancy. Results showed that LA delivered more plasmid DNA to the cytoplasm and achieved transgene expression in more cells than PEI. In contrast, PEI transfected fewer cells but, on average, produced more transgene product per transfected cell. CONCLUSIONS: A comparative transfection model was developed to explain these different characteristics. According to this model, transfection is a multistage process with different transfection agents exerting their primary effect at different stages in this process. This model forecast that it should be possible to prepare a chimeric complex with a transfection efficiency that exceeded that achievable with Lipofectamine or polyethylenimine alone. This prediction was tested and shown to hold for glioma cells, primary astrocytes, and adult neural stems cells. PMID- 15352076 TI - Human FGF-1 gene transfer promotes the formation of collateral vessels and arterioles in ischemic muscles of hypercholesterolemic hamsters. AB - BACKGROUND: Acidic fibroblast growth factor (FGF-1) has been identified as a potent mitogen for vascular cells, inducing formation of mature blood vessels in vitro and in vivo and represents one of the most promising approaches for the treatment of ischemic cardiovascular diseases by gene therapy. Nevertheless, and most probably due to the few experimental models able to address the issue, no study has described the therapeutic effects of FGF-1 gene transfer in subjects with peripheral arterial disease (PAD) exhibiting a clinically relevant cardiovascular pathology. METHODS: In order to assess the potency of FGF-1 gene transfer for therapeutic angiogenesis in ischemic skeletal muscles displaying decreased gene expression levels and sustained impaired formation of collateral vessels and arterioles, we developed a model of PAD in hamsters with a background of hypercholesterolemia. Hamsters fed a cholesterol-rich diet and subjected to hindlimb ischemia exhibit a sustained impaired angiogenic response, as evidenced by decreased angiographic score and histological quantification of arterioles in the ischemic muscles. RESULTS: In this model, we demonstrate that NV1FGF (a human FGF-1 expression plasmid), given intramuscularly 14 days after induction of hindlimb ischemia, promoted the formation of both collateral vessels and arterioles 14 days after treatment (i.e. 28 days post-ischemia). CONCLUSIONS: Our data provide evidence that NV1FGF can reverse the cholesterol-induced impairment of revascularization in a hamster model of hindlimb ischemia by promoting the growth of both collateral vessels and arterioles in ischemic muscles exhibiting significantly decreased levels of gene expression compared with control muscles. Therefore, this study underscores the relevance of NV1FGF gene therapy to overcome perfusion defects in patients with PAD. PMID- 15352079 TI - From cells to tissues: fluorescence confocal microscopy in the study of histological samples. AB - Our knowledge of the genetic mechanisms controlling cell proliferation and differentiation usually originates from in vitro cultured cell line models. However, the definition of the molecular switches involved in control of homeostasis and the understanding of the changes occurring in neoplastic transformation require looking at single cells as the components of a complex tissue network. Histological examination of tissue samples can gain a substantial amount of information from high-resolution fluorescence analysis. In particular, confocal microscopy can help in the definition of functional pathways using multiparameter analysis. In this report, we present acquisition and analysis procedures to obtain high-resolution data from tissue sections. Confocal microscopy coupled to computational restoration, statistical evaluation of spatial correlations, and morphological analysis over large tissue areas were applied to colorectal samples providing a molecular fingerprint of the biological differences inferred from classical histological examination. PMID- 15352080 TI - Optical-sectioning improvement in two-color excitation scanning microscopy. AB - We present a new beam-shaping technique for two-color excitation fluorescence microscopy. We show that by simply inserting a properly designed shaded-ring filter in the illumination beam of smaller wavelength, it is possible to improve the effective optical sectioning capacity of such microscopes by 23%. Such an improvement is obtained at the expense of only a very small increasing of the overall energy in the point-spread-function sidelobes. The performance of this technique is illustrated by a numerical imaging simulation. PMID- 15352081 TI - Colocalization analysis yields superior results after image restoration. AB - Colocalization analysis is a powerful tool for the demonstration of spatial and temporal overlap in the distribution patterns of fluorescent probes. In unprocessed images, background affects image quality by impairing resolution and obscuring image detail in the low-intensity range. Because confocal images suffer from background levels up to 30% maximum intensity, colocalization analysis, which is a typical segmentation process, is limited to high-intensity signal. In addition, noise-induced, false-positive events ("dust") may skew the results. Therefore, suppression of background is crucial for this type of image analysis. Analysis of synthetic and biological objects demonstrates that median filtering is able to eliminate noise-induced colocalization events successfully. Its disadvantages include the occasional generation of false-positive and false negative results as well as the inherent impairment of resolution. In contrast, image restoration by deconvolution suppresses background to very low levels (<10% maximum intensity), which makes additional objects in the low-intensity but high frequency range available for analysis. The improved resolution makes this technique extremely suitable for examination of objects of near resolution size as demonstrated by correlation coefficients. Deconvolution is, however, sensitive to overestimation of the background level. Conclusions for practical application are: (1) In raw images, colocalization analysis is limited to the intensity range above the background level. This means the higher the RS/N the better. Unfortunately, images of most biological specimens have a low RS/N. (2) Filtering improves the result substantially. The reduction of background levels and the concomitant increase of the RS/N are generated at the expense of resolution. This is a quick and simple method in cases where resolution is not a major concern. (3) If colocalization in the low-intensity range and/or maximum resolution play a role, deconvolution should be used. PMID- 15352082 TI - 3D restoration with multiple images acquired by a modified conventional microscope. AB - A problem in high magnification microscopy is the blurring in the imaging of an object. In this article, we demonstrate a restoration technique that simultaneously makes use of the confocal image and the wide-field image. These images can be acquired by a modified conventional microscope. In front of the light-source, there is an array of pinholes. There are no pinholes at the detection plane. Instead, one or more pixels from the CCD camera are used, where the pinholes would have been. Using all pixels gives the wide-field image, but using a selected subset can give a confocal image. The array is used to speed up the process of acquiring the image. Note that the speed of acquisition is proportional to the number of pinholes. We show that the restoration from the two images can lead to a better result than using only one of the images. If this is the case, we show that a distance of 5 times the diameter of the pinholes can give the same results as a distance of 20 times after deconvolution. This offers an increase in acquisition time of a factor 16. PMID- 15352083 TI - Backscattered light confocal imaging of intracellular MTT-formazan crystals. AB - Metabolically active animal and plant cells reduce MTT tetrazolium salt to a corresponding nonfluorescent formazan. Reduction of MTT by viable cells is exploited in a number of tests widely used in biological research. The aim of this study was to optimize a microscopy method of detecting small crystals of MTT formazan formed in intact cells maintained in in vitro cultures. We examined scattering properties of small intracellular crystals of MTT formazan and found that the efficiency of light scattering was dependent on wavelength. Small (<3 microm) crystals of MTT-formazan, formed in viable cells, scattered red, but not blue, light. Large crystals, which are formed later at a stage when cells begin to lose viability, scattered both red and blue light. We conclude that optimal detection of early stages of crystallization of MTT-formazan in living cells is possible using confocal microscopy of red, but not blue, scattered light. High contrast and resolution of images can be achieved by filtering out interference effects in the frequency domain. PMID- 15352084 TI - Saturation modified point spread functions in two-photon microscopy. AB - Excitation saturation can dramatically alter the effective imaging point spread function (PSF) in two-photon fluorescence microscopy. The saturation-modified PSF can have important implications for resolution in fluorescence imaging as saturation leads to both an increased fluorescence observation volume and an altered spatial profile for the PSF. We introduce here a computational approach to accurately quantify molecular excitation profiles that represent the modified imaging PSF in two-photon microscopy under the influence of excitation saturation. An analytical model that accounts for pulsed laser excitation is developed to calculate the influence of saturation at any location within the excitation laser profile. The overall saturation modified molecular excitation profiles are then evaluated numerically. Our results demonstrate that saturation can play an important role in two-photon fluorescence microscopy even with relatively modest excitation levels. PMID- 15352085 TI - Alignment of the cell nucleus from labeled proteins only for 4D in vivo imaging. AB - Studies of protein dynamics by 4D (3D + time) confocal microscopy in vivo are hampered by global cell motion. The time between the acquisitions of the 3D images is in the order of minutes. Therefore, it is not to be expected that the cell as a whole remains fixed in the water basin on the stage. This superimposes a motion on the protein dynamics that has to be removed. We present a robust registration technique to align the cell images that does not require the a priori establishment of point-to-point correspondences. Instead, it uses the distribution of the labeled proteins. After correction for the translation, the 3D rotation of the cell is estimated. A robust intrinsic body coordinate system is constructed via the inertia tensor from the intensity distribution. By combining basis transformation to this intrinsic coordinate system, we can calculated the rotation matrix in a conceptual and computational straightforward manner. We have evaluated the performance of this approach in three experiments with human osteaosarcoma cells (U-2 OS), where the nuclear proteins Histon H4 and PML were visualized. The PML is concentrated in several dozen nuclear spots. Expression of Histon H4 results in a total nuclear staining. The registration results for both channels computed independently are very similar. Practically, this means that only the labeled material needs to be observed and still registration of the cell as a whole can be achieved. PMID- 15352086 TI - Improvement in volume estimation from confocal sections after image deconvolution. AB - The confocal microscope can image a specimen in its natural environment forming a 3D image of the whole structure by scanning it and collecting light through a small aperture (pinhole), allowing in vivo and in vitro observations. So far, the confocal fluorescence microscope (CFM) is considered a true volume imager because of the role of the pinhole that rejects information coming from out-of-focus planes. Unfortunately, intrinsic imaging properties of the optical scheme presently employed yield a corrupted image that can hamper quantitative analysis of successive image planes. By a post-image collection restoration, it is possible to obtain an estimate, with respect to a given optimization criterium, of the true object, utilizing the impulse response of system or Point Spread Function (PSF). The PSF can be measured or predicted so as to have a mathematical and physical model of the image-formation process. Further modelling and recording noise as an additive Gaussian process has used the regularized Iterative Constrained Tykhonov Miller (ICTM) restoration algorithm for solving the inverse problem. This algorithm finds the best estimate iteratively searching among the possible positive solutions; in the Fourier domain, such an approach is relatively fast and elegant. In order to compare the effective improvement in the quantitative image information analysis, we measured the volume of reference objects before and after image restoration, using the isotropic Fakir method. PMID- 15352087 TI - Adaptive image-processing technique and effective visualization of confocal microscopy images. AB - A common observation about confocal microscopy images is that lower image stacks have lower voxel intensities and are usually blurred in comparison with the upper ones. The key reasons are light absorption and scattering by the objects and particles in the volume through which light passes. This report proposes a new technique to reduce such noise impacts in terms of an adaptive intensity compensation and structural sharpening algorithm. With these image-processing procedures, effective 3D rendering techniques can be applied to faithfully visualize confocal microscopy data. PMID- 15352088 TI - Automated acquisition and processing of multidimensional image data in confocal in vivo microscopy. AB - The successful development of visualization techniques for live cell imaging leads to the development of suitable software for the acquisition and processing of multidimensional image data. This report compares several possible approaches to image acquisition and processing in confocal in vivo microscopy and suggests new alternatives to the published methods. Special attention is paid to spinning disk systems based either on a classical Nipkow disk or on the microlens principle. This study shows how to optimize image acquisition process in live cell studies using camera binning feature and how to perform object tracking using a new fast image registration method based on the graph theory. PMID- 15352089 TI - AFM/CLSM data visualization and comparison using an open-source toolkit. AB - There is a vast difference in the traditional presentation of AFM data and confocal data. AFM data are presented as surface contours while confocal data are usually visualized using either surface- or volume-rendering techniques. Finding a common meaningful visualization platform is not an easy task. AFM and CLSM technologies are complementary and are more frequently being used to image common biological systems. In order to provide a presentation method that would assist us in evaluating cellular morphology, we propose a simple visualization strategy that is comparative, intuitive, and operates within an open-source environment of ImageJ, SurfaceJ, and VolumeJ applications. In order to find some common ground for AFM-CLSM image comparison, we have developed a plug-in for ImageJ, which allows us to import proprietary image data sets into this application. We propose to represent both AFM and CLSM image data sets as shaded elevation maps with color-coded height. This simple technique utilizes the open source VolumeJ and SurfaceJ plug-ins. To provide an example of this visualization technique, we evaluated the three-dimensional architecture of living chick dorsal root ganglia and sympathetic ganglia measured independently with AFM and CLSM. PMID- 15352090 TI - Applying spectral fingerprinting to the analysis of FRET images. AB - Forster resonance energy transfer (FRET) allows one to study interactions between two fluorescently labeled molecules (donors and acceptors) at distances on the order of 5 nm. Many studies have described methods of how to measure the efficiency of FRET. However, few have addressed the question of how fluorescence from unpaired donors and acceptors can be determined in addition to that from FRET-pairs and how the signal-to-noise ratio (SNR) of such estimates depends on the presence of the partner species. Such knowledge, however, is essential for many biological applications, in which-after initial characterization of the spectral properties of a well-defined donor-acceptor complex-the in vivo affinity and stoichiometry of the complex is of interest. Here, we provide a theoretical analysis on how spectral fingerprinting can be applied to separate fluorescence of FRET pairs from that originating from unpaired donors and acceptors and how to select imaging parameters to optimize the SNR of the estimates. Thereby, we uncover a fundamental problem in this application and discuss ways to evade its adverse consequences. We compare the expected resolution of traditional FRET measures with that of optimized spectral fingerprinting and analyze the resolution of a method for FRET measurements that combines spectral with fluorescence lifetime information. PMID- 15352091 TI - Three-dimensional microscopy migrates to the web with "PowerUp Your Microscope". AB - "PowerUp Your Microscope" is a software package designed and realized for the optimization of 3D optical microscopy image quality using the Internet and inverse problems computational approaches. The package is mainly devoted to 3D microscopy users, being operative for wide-field, confocal, and multiphoton microscopy. It provides the microscopy community with an extremely easy and comparatively powerful access to advanced image restoration methods. The core of the computational section is the optical system modeling and inverse deconvolution implementation, which is strongly linked to Web-based software and technology. This project constitutes a real and effective migration to the Web, extending computational approaches to image restoration to the whole microscopy user community, regardless of their background. PMID- 15352092 TI - Cluster computing for digital microscopy. AB - Microscopy is becoming increasingly digital and dependent on computation. Some of the computational tasks in microscopy are computationally intense, such as image restoration (deconvolution), some optical calculations, image segmentation, and image analysis. Several modern microscope technologies enable the acquisition of very large data sets. 3D imaging of live cells over time, multispectral imaging, very large tiled 3D images of thick samples, or images from high throughput biology all can produce extremely large images. These large data sets place a very large burden on laboratory computer resources. This combination of computationally intensive tasks and larger data sizes can easily exceed the capability of single personal computers. The large multiprocessor computers that are the traditional technology for larger tasks are too expensive for most laboratories. An alternative approach is to use a number of inexpensive personal computers as a cluster; that is, use multiple networked computers programmed to run the problem in parallel on all the computers in the cluster. By the use of relatively inexpensive over-the-counter hardware and open source software, this approach can be much more cost effective for many tasks. We discuss the different computer architectures available, and their advantages and disadvantages. PMID- 15352094 TI - Control of molecular topology and metal nuclearity in multimetallic assemblies: designer metallosiloxanes derived from silanetriols. AB - Lipophilic N-bonded silanetriol RSi(OH)(3) (R=(2,6-iPr(2)C(6)H(3))N(SiMe(3))) can be utilized as an effective synthon for building a variety of multimetallic assemblies containing the Si-O-M motif. The type of metallosiloxane synthesized its nuclearity and its molecular topology-can be readily modulated by the choice of the metal substrate, reaction stoichiometry, and reaction conditions. It is anticipated that the synthetic principles elaborated here will allow the design of many other multifunctional synthons. PMID- 15352095 TI - Galactose oxidase models: solution chemistry, and phenoxyl radical generation mediated by the copper status. AB - Galactose oxidase (GO) is an enzyme that catalyzes two-electron oxidations. Its active site contains a copper atom coordinated to a tyrosyl radical, the biogenesis of which requires copper and dioxygen. We have recently studied the properties of electrochemically generated mononuclear Cu(II)-phenoxyl radical systems as model compounds of GO. We present here the solution chemistry of these ligands under various copper and dioxygen statuses: N(3)O ligands first chelate Cu(II), leading, in the presence of base, to [Cu(II)(ligand)(CH(3)CN)](+) complexes (ortho-tert-butylated ligands) or [(Cu(II))(2)(ligand)(2)](2+) complexes (ortho-methoxylated ligands). Excess copper(II) then oxidizes the complex to the corresponding mononuclear Cu(II)-phenoxyl radical species. N(2)O(2) tripodal ligands, in the presence of copper(II), afford directly a copper(II)-phenoxyl radical species. Addition of more than two molar equivalents of copper(II) affords a Cu(II)-bis(phenoxyl) diradical species. The donor set of the ligand directs the reaction towards comproportionation for ligands possessing an N(3)O donor set, while disproportionation is observed for ligands possessing an N(2)O(2) donor set. These results are discussed in the light of recent results concerning the self-processing of GO. A path involving copper(II) disproportionation is proposed for oxidation of the cross-linked tyrosinate of GO, supporting the fact that both copper(I) and copper(II) activate the enzyme. PMID- 15352096 TI - Chiral recognition by resorcin[4]arene receptors: intrinsic kinetics and dynamics. AB - Molecular recognition of representative amino acids (A) by a chiral amido[4]resorcinarene receptor (1(L)) was investigated in the gas phase by ESI-FT ICR mass spectrometry. The ligand displacement reaction between noncovalent diastereomeric [1(L).H.A](+) complexes and the 2-aminobutane enantiomers (B) exhibits a distinct enantioselectivity with regard to both the leaving amino acid A and the amine reactant B. The emerging selectivity picture, discussed in the light of molecular mechanics and molecular dynamics calculations, points to chiral recognition by 1(L), as determined by the effects of the host asymmetric frame on the structure, stability, and rearrangement dynamics of the diastereomeric [1(L).H.A](+) complexes and the orientation of the amine reactant B in encounters with [1(L).H.A](+). The results contribute to the development of a dynamic model of chiral recognition of biomolecules by enzyme mimics in the unsolvated state. PMID- 15352097 TI - Synthesis of a polymer-bound galactosylamine and its application as an immobilized chiral auxiliary in stereoselective syntheses of piperidine and amino acid derivatives. AB - A 2,3,4-tri-O-pivaloylated beta-D-galactopyranosyl azide bearing a hydroxy functionalized spacer unit at the C-6 position of the galactose was synthesized and immobilized on the solid phase by using a polymer-bound chlorosilane. The azide was reduced to the corresponding galactopyranosylamine, which served as a versatile chiral auxiliary in highly diastereoselective Ugi four-component condensation reactions at ambient temperature. Fluoride-induced cleavage from the polymeric support furnished N-glycosylated N-acylated alpha-amino acid amides. The reaction of the immobilized galactosylamine with aldehydes gave rise to the corresponding aldimines, which underwent a domino Mannich-Michael condensation reaction with Danishefsky's diene at ambient temperature to yield 2-substituted 5,6-didehydropiperidin-4-ones on the solid phase. Subsequent cleavage with tetra n-butylammonium fluoride delivered the N-glycosylated products in high yields, purities, and diastereoselectivities. A chemoselective 1,4-hydride addition to the polymer-bound dehydropiperidinones was achieved in the presence of the bulky oxygenophilic Lewis acid methylaluminum [bis(2,6-di-tert-butyl-4 methylphenoxide)]. The conjugate addition of cyano-modified Gilman reagents to the immobilized dehydropiperidinones furnished 2,6-cis-substituted piperidine derivatives as the major diastereomers that were isolated after cleavage from the support. PMID- 15352098 TI - Biomimetic synthesis of the tumor-associated (2,3)-sialyl-T antigen and its incorporation into glycopeptide antigens from the mucins MUC1 and MUC4. AB - Glycoproteins on epithelial tumor cells often exhibit aberrant glycosylation profiles. The incomplete formation of the glycan side chains resulting from a down-regulated glucosamine transfer and a premature sialylation results in additional peptide epitopes, which become accessible to the immune system in mucin-type glycoproteins. These cancer-specific structure alterations are considered to be a promising basis for selective immunological attack on tumor cells. Among the tumor-associated saccharide antigens, the (2,3)-sialyl-T antigen has been identified as the most abundant glycan, found in several different carcinoma cell lines. According to a linear biomimetic strategy, the (2,3)-sialyl T antigen was synthesized by a stepwise glycan chain extension of a protected galactosamine-threonine precursor. For the construction of immunostimulating antigens combining both peptide and saccharide motifs, this antigen was incorporated into glycopeptide partial structures from the mucins MUC1 and MUC4 by sequential solid-phase synthesis. PMID- 15352099 TI - De novo metallonucleases based on helix-loop-helix motifs. AB - Three new 42-mer peptides (PR I-III) designed to fold into a hairpin helix-loop helix motif have been prepared. In the peptide sequence two (PR II-III) or four (PR I) copies of an unnatural amino acid bearing a triazacyclononane metal-ion binding site (ATANP) have been inserted in appropriate positions to allow the ligand subunits to face each other either within the same helix or between the two helices of the hairpin motif. Circular dichroism (CD) studies in solution have shown that the apopeptides adopt a well-defined helix-loop-helix tertiary structure that dimerizes in solution at concentrations above 200 microM to form a four-helix bundle. However, the helical content is strongly dependent on pH and metal-ion binding. Both protonation of the amines of the triazacyclononane units present in the ATANP lateral arm and complexation with Zn(II) ions cause a significant decrease of the helical content of the sequences. The Zn(II) complexes of the three peptides catalyze the transesterification of the RNA model substrate 2-hydroxypropyl-p-nitrophenyl phosphate (HPNP) with different efficiency. The best catalyst appears to be PR I-4 Zn(II), that is, the peptide incorporating four ATANP units. Michaelis-Menten saturation kinetics allowed us to estimate that substrate fully bound to the catalyst reacts 380 times faster than in its absence. The kinetic evidence suggests cooperativity between (at least two) metal ions: one activating the nucleophilic species (directly or indirectly) and the other facilitating nucleophilic attack by coordination of the phosphate. PMID- 15352100 TI - Stereoselective synthesis of (+)-nephrosteranic acid, (+)-trans-cognac lactone, and (+)-trans-whisky lactone using a chiral cyclohexadienyl Ti compound. AB - We present the stereoselective transfer of cyclohexadienyl from 3-metalated 1,4 cyclohexadienes to various aldehydes. Lewis-acid-mediated "allylation" of aldehydes by treatment with 3-silylated and 3-stannylated 1,4-cyclohexadienes could not be achieved with high diastereoselectivity. In contrast, cyclohexadienyl titanium compounds reacted with both aliphatic and aromatic aldehydes with good-to-excellent diastereoselectivities. Reaction of a chiral TADDOL-derived (TADDOL, 2,2-dimethyl-alpha,alpha,alpha',alpha'-tetraphenyl-1,3 dioxolandimethanol) cyclohexadienyl Ti derivative with various aldehydes led to the corresponding homoallylic alcohols with excellent diastereo- and enantioselectivities. Lower selectivities were obtained with chiral B cyclohexadienyldiisopinocampheylborane. The 1,3-cyclohexadienes are very useful building blocks for the preparation of biologically important gamma butyrolactones. Short efficient syntheses of (+)-nephrosteranic acid, (+)-trans whisky lactone, and (+)-trans-cognac lactone by desymmetrization of 1,4 cyclohexadiene are described. PMID- 15352101 TI - Synthesis and characterization of pentachlorophenyl-metal derivatives with d0 and d10 electron configurations. AB - By reaction of [TiCl(3)(thf)(3)] with LiC(6)Cl(5), the homoleptic organotitanium(III) derivative [Li(thf)(4)][Ti(III)(C(6)Cl(5))(4)] (1) has been prepared as a paramagnetic (d(1), S = 1/2, g(av) = 1.959(2)), extremely air sensitive compound. Oxidation of 1 with [N(C(6)H(4)Br-4)(3)][SbCl(6)] gives the diamagnetic (d(0)) organotitanium(IV) species [Ti(IV)(C(6)Cl(5))(4)] (2). Compounds 1 and 2 are also electrochemically related (E(1/2) = 0.05 V). The homoleptic, diamagnetic (d(10)) compounds [N(PPh(3))(2)][Tl(C(6)Cl(5))(4)] (3) and [Sn(C(6)Cl(5))(4)] (4) have also been prepared. Nearly tetrahedral environments have been found for the d(0), d(10), and d(1) metal centers in the molecular structures of compounds 2-4 as well as in that of [Li(thf)(2)(OEt(2))(2)][Ti(III)(C(6)Cl(5))(4)].CH(2)Cl(2) (1') (X-ray diffraction). The reaction of the heavier Group 4 metal halides, MCl(4) (M = Zr, Hf) with LiC(6)Cl(5) in the presence of [NBu(4)]Br gives, in turn, the heteroleptic species [NBu(4)][M(C(6)Cl(5))(3)Cl(2)] (M = Zr (5), Hf (6)). Compounds 5 and 6 are isomorphous and isostructural, with the metal center in a trigonal-bipyramidal (TBPY-5) environment defined by two axial Cl ligands and three equatorial C(6)Cl(5) groups (X-ray diffraction). No redox features are observed for compounds 3-6 in CH(2)Cl(2) solution between -1.6 and +1.6 V. PMID- 15352102 TI - Olefins as steering ligands for homogeneously catalyzed hydrogenations. AB - Iridium(I) complexes containing a (5H-dibenzo[a,d]cyclohepten-5-yl)-phosphane (tropp(R); R = phosphorus-bound substituent = Ph, Cyc) as a rigid, concave shaped, mixed phosphane olefin ligand were prepared and tested as catalyst precursors in the hydrogenation of imines. With the complex [Ir(tropp(Cyc))(cod)]OTf, turnover frequencies (TOFs) of >6000 h(-1) were reached in the hydrogenation of N-phenyl-benzylidenamine, PhN==CHPh. Lower activities (TOF>80 h(-1)) are observed with N-phenyl-(1-phenylethylidene)amine, PhN==CMePh. Chiral tropp-type ligands were prepared in few simple steps. Monosubstitution of the olefinic unit in the dibenzo[a,d]cycloheptenyl moiety with (R)- or (S) mentholate gave mixtures of diastereomers that could be separated and isolated in enantiomerically pure form. Iridium(I) complexes with these ligands are rare examples of side-on bonded enolether complexes. In catalytic imine hydrogenations, complete conversion (>98 %) was reached in all cases (conditions: p[H(2)] = 50 bar, T = 50 degrees C, t = 2 h, substrate/catalyst 100:1). The best enantiomeric excess (ee = 86 % S isomer) was reached with PhN==CMePh as substrate and the R,R form of the (10-menthyloxy-5H-dibenzo[a,d]cyclohepten-5 yl)diphenylphosphane ligand. The iridium(I) complex containing the same phosphane gave a 60 % ee (S isomer) with the enamide N-(1-phenylvinyl)acetamide as substrate (conditions: p[H(2)] = 4 bar, T = 50 degrees C, t = 18 h, substrate/catalyst = 50:1). These reactions constitute the first examples in which chiral olefins have been used as steering ligands in catalytic enantioselective hydrogenations. PMID- 15352103 TI - Bis(pyridine)iodonium tetrafluoroborate (IPy2BF4): a versatile oxidizing reagent. AB - The use of bis(pyridine)iodonium tetrafluoroborate (IPy(2)BF(4)) as an oxidizing agent towards different types of alcohols is reported. The observed reactivity involves different reaction pathways, as a function both of the structures of the starting materials and of the experimental conditions. Interestingly, the title iodine-containing compound is capable of a tuneable reaction with simple cycloalkanols, providing straight and selective access either to omega iodocarbonyl compounds or to ketones, a previously unreported and chemoselective range of oxidation potential. Furthermore, appropriate conditions for the preparation of aldehydes and esters from primary alcohols by easily performed experimental procedures were also established. The beta-scission reactions of cycloalkanols and the alpha-oxidation processes of primary, secondary and benzylic alcohols are discussed. PMID- 15352104 TI - Binding H2, N2, H-, and BH3 to transition-metal sulfur sites: synthesis and properties of [RuL(PR3)(N2Me2S2)] Complexes (L=eta2-H2, H-, BH3; R=Cy, iPr). AB - The reactions of [Ru(N(2))(PR(3))('N(2)Me(2)S(2)')] ['N(2)Me(2)S(2)'=1,2 ethanediamine-N,N'-dimethyl-N,N'-bis(2-benzenethiolate)(2-)] [1 a (R=iPr), 1 b (R=Cy)] and [micro-N(2)[Ru(N(2))(PiPr(3))('N(2)Me(2)S(2)')](2)] (1 c) with H(2), NaBH(4), and NBu(4)BH(4), intended to reduce the N(2) ligands, led to substitution of N(2) and formation of the new complexes [Ru(H(2))(PR(3))('N(2)Me(2)S(2)')] [2 a (R=iPr), 2 b (R=Cy)], [Ru(BH(3))(PR(3))('N(2)Me(2)S(2)')] [3 a (R=iPr), 3 b (R=Cy)], and [Ru(H)(PR(3))('N(2)Me(2)S(2)')](-) [4 a (R=iPr), 4 b (R=Cy)]. The BH(3) and hydride complexes 3 a, 3 b, 4 a, and 4 b were obtained subsequently by rational synthesis from 1 a or 1 b and BH(3).THF or LiBEt(3)H. The primary step in all reactions probably is the dissociation of N(2) from the N(2) complexes to give coordinatively unsaturated [Ru(PR(3))('N(2)Me(2)S(2)')] fragments that add H(2), BH(4) (-), BH(3), or H(-). All complexes were completely characterized by elemental analysis and common spectroscopic methods. The molecular structures of [Ru(H(2))(PR(3))('N(2)Me(2)S(2)')] [2 a (R=iPr), 2 b (R=Cy)], [Ru(BH(3))(PiPr(3))('N(2)Me(2)S(2)')] (3 a), [Li(THF)(2)][Ru(H)(PiPr(3))('N(2)Me(2)S(2)')] ([Li(THF)(2)]-4 a), and NBu(4)[Ru(H)(PCy(3))('N(2)Me(2)S(2)')] (NBu(4)-4 b) were determined by X-ray crystal structure analysis. Measurements of the NMR relaxation time T(1) corroborated the eta(2) bonding mode of the H(2) ligands in 2 a (T(1)=35 ms) and 2 b (T(1)=21 ms). The H,D coupling constants of the analogous HD complexes HD-2 a ((1)J(H,D)=26.0 Hz) and HD-2 b ((1)J(H,D)=25.9 Hz) enabled calculation of the H- D distances, which agreed with the values found by X-ray crystal structure analysis (2 a: 92 pm (X-ray) versus 98 pm (calculated), 2 b: 99 versus 98 pm). The BH(3) entities in 3 a and 3 b bind to one thiolate donor of the [Ru(PR(3))('N(2)Me(2)S(2)')] fragment and through a B-H-Ru bond to the Ru center. The hydride complex anions 4 a and 4 b are extremely Bronsted basic and are instantaneously protonated to give the eta(2)-H(2) complexes 2 a and 2 b. PMID- 15352105 TI - Adrenaline recognition in water. AB - Host molecule 1 displays a high affinity in water towards catecholamines and especially related structures such as beta-blockers with extended aromatic pi faces (up to 7x10(3) M(-1) for each single complexation step or 5x10(7) M(-2) for both steps). The amphiphilic structural design leads to an extensive self association of host molecules through their aromatic flanks. Above a cmc (critical micelle concentration) of 3x10(-4) M, host 1 forms micelles that produce a favorable microenvironment for hydrophobic interactions with the included guest molecules. Electrostatic attraction of the ammonium alcohol by the phosphonate anions is thus combined with hydrophobic contributions between the aromatic moieties. Ionic hydrogen bonds with polar OH or NH groups of the guest enforce the non-covalent interactions, and finally lead to increased specificity. Both its affinity and its selectivity towards adrenergic receptor substrates are greatly enhanced if the receptor molecule 1 is transferred from water into a lipid monolayer. Catecholamines and beta-blockers lead to drastically different effects at concentrations approaching the micromolar regime. Especially beta blockers with minute structural changes can be easily distinguished from each other. In both cases, extensive hydrophobic interactions with a self-associated and/or self-organized microenvironment are largely responsible for the observed high efficiency and specificity. PMID- 15352106 TI - Relative binding affinities of molecular capsules investigated by ESI-mass spectrometry. AB - ESI-MS(/MS) has been used as a method which allows the fast, unambiguous and sensitive simultaneous detection and relative stability approximation of supramolecular assemblies in mixtures. In spite of the obvious fundamental differences between solution and gas phase, ESI-MS in the case of self-assembled molecular capsules has been shown to produce very similar results to single binding experiments monitored by NMR titrations as well as conformational searches performed by Monte-Carlo simulations. MS/MS experiments reveal the same relative order of gas phase stabilities as previously found in solution. Moreover, proton transfer reactions which lead to new molecular capsules, are not detectable in the time-averaged NMR spectrum. However, the newly produced species are found in the complex mixtures by ESI-MS and can be conveniently characterized by subsequent MS/MS experiments: in a collision-induced dissociation the single half-spheres are easily discovered and structurally assigned. Thus, ESI-MS has worked as a valuable tool for the rapid screening of complex supramolecular mixtures and in combination with MS/MS experiments elucidated both the path of unexpected side reactions as well as the thermodynamic gas-phase stabilities of all components in the mixture. PMID- 15352107 TI - Hydrogen-bonding cooperativity: using an intramolecular hydrogen bond to design a carbohydrate derivative with a cooperative hydrogen-bond donor centre. AB - Neighbouring groups can be strategically located to polarise HO.OH intramolecular hydrogen bonds in an intended direction. A group with a unique hydrogen-bond donor or acceptor character, located at hydrogen-bonding distance to a particular OH group, has been used to initiate the hydrogen-bond network and to polarise a HO.OH hydrogen bond in a predicted direction. This enhanced the donor character of a particular OH group and made it a cooperative hydrogen-bond centre. We have proved that a five-membered-ring intramolecular hydrogen bond established between an amide NH group and a hydroxy group (1,2-e,a), which is additionally located in a 1,3-cis-diaxial relationship to a second hydroxy group, can be used to select a unique direction on the six-membered-ring intramolecular hydrogen bond between the two axial OH groups, so that one of them behaves as an efficient cooperative donor. Talose derivative 3 was designed and synthesised to prove this hydrogen bonding network by NMR spectroscopy, and the mannopyranoside derivatives 1 and 2 were used as models to demonstrate the presence in solution of the 1,2-(e,a)/five membered-ring intramolecular hydrogen bond. Once a well-defined hydrogen-bond is formed between the OH and the amido groups of a pyranose ring, these hydrogen bonding groups no longer act as independent hydrogen-bonding centres, but as hydrogen-bonding arrays. This introduces a new perspective on the properties of carbohydrate OH groups and it is important for the de novo design of molecular recognition processes, at least in nonpolar media. Carbohydrates 1-3 have shown to be efficient phosphate binders in nonpolar solvents owing to the presence of cooperative hydroxy centres in the molecule. PMID- 15352108 TI - Diastereoselective remote C-H activation by hydroboration. AB - Hydroboration of tetrasubstituted or trisubstituted alkenes with BH(3) and subsequent thermolysis allows remote diastereoselective C-H activation of neighboring aryl rings. Tetrasubstituted and trisubstituted 1,1-diphenylethylene derivatives undergo a highly stereoselective 1,2-rearrangement followed by remote C-H activation to lead, after oxidative workup, to a diol in which the relative stereochemistry of two stereocenters has been controlled. The mechanism of this remote activation has been studied and extended to related molecules that undergo this stereoselective C-H activation, namely alkenylbiphenyl systems or alkenes with only one phenyl ring, such as alkenylbenzenes, or bicyclic systems. We have shown that this reaction allows diastereoselective synthesis of molecules with up to three contiguous chiral centers. PMID- 15352109 TI - Synthesis of tailor-made glycoconjugate mimetics of heparan sulfate that bind IFN gamma in the nanomolar range. AB - We have recently described the preparation of three building blocks for the combinatorial synthesis of heparan sulfate (HS) fragments. Herein we show that one of these building blocks (disaccharide 4) allows the preparation, in high yields and with total alpha stereoselectivity, of tetra-, hexa- and octasaccharides from the heparin (HP) regular region, by using 2+2, 2+4 and 4+4 glycosylation strategies, respectively. These oligosaccharides were processed into sulfated derivatives bearing an allyl moiety in the anomeric position. The UV-promoted conjugation of these compounds with alpha,omega bis(thio)poly(ethylene glycol) spacers of three different lengths allowed us to prepare nine benzylated glycoconjugates. After final deprotection, the glycoconjugates 1 a-c, 2 a-c and 3 a-c were obtained and their ability to inhibit the interaction between IFN-gamma and HP was tested by using surface plasmon resonance detection. Compound 3 b, containing two HP octasaccharides linked by a 50-A linker was able to inhibit the IFN-gamma/HP interaction with an IC(50) value of approximately 35 nM. In addition, the nine glycoconjugates were perfect tools in the study to ascertain the topology of the IFN-gamma binding site on HS. Compounds 1 a-c, 2 a-c and 3 a-c, by mimicking the alternating sulfated and nonsulfated regions found in HS, thus comprise the first example of a library of synthetic HS mimetics giving access to the "second level of molecular diversity" found in HS. PMID- 15352110 TI - A new color of the synthetic chameleon methoxyallene: synthesis of trifluoromethyl-substituted pyridinol derivatives: an unusual reaction mechanism, a remarkable crystal packing, and first palladium-catalyzed coupling reactions. AB - Addition of lithiated methoxyallene to pivalonitrile afforded after aqueous workup the expected iminoallene 1 in excellent yield. Treatment of this intermediate with silver nitrate accomplished the desired cyclization to the electron-rich pyrrole derivative 2 in moderate yield. Surprisingly, trifluoroacetic acid converted iminoallene 1 to a mixture of enamide 3 and trifluoromethyl-substituted pyridinol 4 (together with its tautomer 5). A plausible mechanism proposed for this intriguing transformation involves addition of trifluoroacetate to the central allene carbon atom of an allenyl iminium intermediate as crucial step. Enamide 3 is converted to pyridinol 4 by an intramolecular aldol-type process. A practical direct synthesis of trifluoromethyl-substituted pyridinols 4, 10, 11, and 12 starting from typical nitriles and methoxyallene was established. Pyridinol 10 shows an interesting crystal packing with three molecules in the elementary cell and a remarkable helical supramolecular arrangement. Trifluoromethyl-substituted pyridinol 4 was converted to the corresponding pyridyl nonaflate 13, which is an excellent precursor for palladium-catalyzed reactions leading to pyridine derivatives 14-16 in good to excellent yields. The new synthesis of trifluoromethyl-substituted pyridines disclosed here demonstrates a novel reactivity pattern of lithiated methoxyallene which is incorporated into the products as the unusual tripolar synthon B. PMID- 15352111 TI - Using noncovalent intra-strand and inter-strand interactions to prescribe helix formation within a metallo-supramolecular system. AB - The effect of inter-strand and intra-strand interactions is explored in a metallo supramolecular system in which the metal-ligand coordination requirements may be satisfied by more than one different supramolecular architecture. This is achieved by introducing alkyl substituents onto the spacers of readily prepared bis(pyridylimine) ligands. The alkyl substituents induce twisting within the ligand strand and this intra-strand effect favours formation of helical architectures. The alkyl substituents also introduce inter-strand CH.pi interactions into the system. For the smaller methyl group these are most effectively accommodated in a trinuclear circular helicate architecture. A solution mixture of dinuclear double-helicate and trinuclear circular helicate results from which, for copper(I), the trinuclear circular helicate crystallises. The CH.pi interactions endow the circular helicate with a bowl-shaped conformation and the triangular unit aggregates into a tetrahedral ball-shaped array. Low-temperature NMR studies indicate that the CH.pi interactions also confer a bowl-shaped conformation on the triangle in solution. The larger ethyl groups can sustain intra-strand CH.pi interactions in the lower nuclearity double helical system and this is the unique architecture for that ligand system in both solution and the solid state. Crystal structures are described for both the copper(I) and silver(I) complexes. Thus we show that intra-strand interactions may be used to induce helicity within this system, while the nuclearity of the array can be prescribed by the inter-strand interactions. PMID- 15352112 TI - Generation of cationic [Zr-[tert-butyl enolate]] reactive species: methyl abstraction versus hydride abstraction. AB - Treatment of the neutral methyl-Zr-enolate [Cp(2)Zr(Me)[O(tBuO)C=CMe(2)]] (1) with one equivalent of B(C(6)F(5))(3) or [HNMe(2)Ph][B(C(6)F(5))(4)] as a methyl abstractor in THF at 0 degrees C leads to the selective formation of the free ion pair complex [Cp(2)Zr(THF)[O(tBuO)C=CMe(2)]](+) [anion](-) (2) (anion=MeB(C(6)F(5))(3) (-), B(C(6)F(5))(4) (-)), which is relevant to the controlled polymerization of methacrylates. Cation 2 rapidly decomposes at 20 degrees C in THF with release of one equivalent of isobutene to form the cationic Zr-carboxylate species [Cp(2)Zr(THF)(O(2)CiPr)](+) (3), through a proposed intramolecular proton transfer process from the tert-butoxy group to the enolate. The reaction of 1 with one equivalent of B(C(6)F(5))(3) or [HNMe(2)Ph][B(C(6)F(5))(4)] in CH(2)Cl(2) leads to the direct, rapid formation of the dimeric micro-isobutyrato-Zr dicationic species [[Cp(2)Zr[micro (O(2)CiPr)]](2)](2+) (4), which gives 3 upon dissolution in THF. Contrastingly, when [Ph(3)C][B(C(6)F(5))(4)] is used to generate the cationic Zr-enolate species from 1 in CD(2)Cl(2), a 15:85 mixture of dicationic complexes 4 and [[Cp(2)Zr[micro-(O(2)C-C(Me)=CH(2))]](2)](2+)[B(C(6)F(5))(4)]]2-(5 [B(C(6)F(5))(4)](2)) is obtained quantitatively. The formation of 5 is proposed to arise from initial hydride abstraction from a methyl enolate group by Ph(3)C(+), as supported by the parallel production of Ph(3)CH, and subsequent elimination of methane and isobutene. In addition to standard spectroscopic and analytical characterizations for the isolated complexes 2-5, complexes 4 and 5 have also been structurally characterized by X-ray diffraction studies. PMID- 15352113 TI - Cubane-type Mo3CoS4 molecular clusters with three different metal electron populations: structure, reactivity and their use in the synthesis of hybrid charge-transfer salts. AB - Heterodimetallic cubane-type complexes coordinated to diphosphanes [Mo(3)CoS(4)(dmpe)(3)Cl(4)](+) ([1](+)) (dmpe=1,2-bis(dimethylphosphanyl)ethane), [Mo(3)CoS(4)(dmpe)(3)Cl(4)] (1) and [Mo(3)CoS(4)(dmpe)(3)Cl(3)(CO)] (2) with 14, 15 and 16 metal electrons, respectively, have been prepared from the [Mo(3)S(4)(dmpe)(3)Cl(3)](+) trinuclear precursor using [Co(2)(CO)(8)] or CoCl(2) as cobalt source. Cluster complexes [1](+) and 1 are easily interconverted chemically and electrochemically. The Co-Cl distance increases upon electron addition and substitution of the chlorine atom coordinated to cobalt with CO only takes place in presence of a reducing agent to give complex 2. Structural changes in the intermetallic distances agree with the entering electrons occupying an orbital which is basically Mo-Mo non-bonding and slightly Mo-Co bonding. Magnetic susceptibility measurements for [1](+) and 1 are consistent with the presence of two and one unpaired electrons, respectively and therefore with an "e" character for the HOMO orbital. Oxidation of 1 with TCNQ results in the formation of a charge transfer salt formulated as [1](+)[TCNQ](-) with alternate layers of paramagnetic cluster cations and also paramagnetic organic anions. There is no magnetic interaction between layers and the thermal variation of the magnetic susceptibility has been modelled as a S= 1/2 TCNQ antiferromagnetic chain plus a S=1 cluster monomer with zero field splitting. PMID- 15352114 TI - Cylindrical micelles from the aqueous self-assembly of an amphiphilic poly(ethylene oxide)-b-poly(ferrocenylsilane) (PEO-b-PFS) block copolymer with a metallo-supramolecular linker at the block junction. AB - A supramolecular AB diblock copolymer has been prepared by the sequential self assembly of terpyridine end-functionalized polymer blocks by using Ru(III)/Ru(II) chemistry. By this synthetic strategy a hydrophobic poly(ferrocenylsilane) (PFS) was attached to a hydrophilic poly(ethylene oxide) (PEO) block to give an amphiphilic metallo-supramolecular diblock copolymer (PEO/PFS block ratio 6:1). This compound was used to form micelles in water that were characterized by a combination of dynamic and static light scattering, transmission electron microscopy, and atomic force microscopy. These complementary techniques showed that the copolymers investigated form rod-like micelles in water; the micelles have a constant diameter but are rather polydisperse in length, and light scattering measurements indicate that they are flexible. Crystallization of the PFS in these micelles was observed by differential scanning calorimetry, and is thought to be the key behind the formation of rod-like structures. The cylindrical micelles can be cleaved into smaller rods whenever the temperature of the solution is increased or they are exposed to ultrasound. PMID- 15352115 TI - Theoretical study of the electrocyclic ring closure of hydroxypentadienyl cations. AB - At the 6-311G* level of theory, DFT methods predict that the rearrangement of 1,4 dihydroxy-5-methylpentadienyl cation 1 (R = Me) to protonated trans-3-hydroxy-2 methylcyclopent-4-en-1-one 2, an intermediate step in the Piancatelli reaction or rearrangement of furfuryl carbinols to trans-2-alkyl(aryl)-3-hydroxycyclopent-4 en-1-one, is a concerted electrocyclic process. Energetic, magnetic, and stereochemical criteria are consistent with a conrotatory electrocyclic ring closure of the most stable out,out-1 isomer to afford trans-2. Although the out,in-1 isomer is thermodynamically destabilized by 6.84 kcal mol(-1), the activation energy for its cyclization is slightly lower (5.29 kcal mol(-1) versus 5.95 kcal mol(-1)). The cyclization of the isomers of 1 with the C1-hydroxy group inwards showed considerably higher activation energies than their outwards counterparts. in,out-1, although close in energy to out,out-1 (difference of 1.57 kcal mol(-1)) required about 10 kcal mol(-1) more to reach the corresponding transition structure. The value measured for the activation energy of in,in-1 (17.32 kcal mol(-1)) eliminates the alternative conrotatory electrocyclization of this isomer to provide trans-2. Geometric scrambling by isomerization of the terminal C1--C2 bond of 1 is also unlikely to compete with electrocyclization. The possibility to interpret the 1-->2 reaction as a nonpericyclic cationic cyclization was also examined through NBO analysis, and the study of bond lengths and atomic charges. It was found that the 1-->2 concerted rearrangement benefits from charge separation at the cyclization termini, an effect not observed in related concerted electrocyclic processes, such as the classical Nazarov reaction 3-->4 or the cyclization of the isomeric 2-hydroxypentadienyl cation 5. PMID- 15352116 TI - Solid-phase synthesis of dihydrovirginiamycin S1, a streptogramin B antibiotic. AB - We describe the first solid-phase synthesis of dihydrovirginiamycin S(1), a member of the streptogramin B family of antibiotics, which are nonribosomal peptide natural products produced by Streptomyces. These compounds, along with the synergistic group A components, are "last line of defense" antimicrobial agents for the treatment of life-threatening infections such as vancomycin resistant enterococci. The synthesis features an on-resin cyclization and is designed to allow production of streptogramin B analogues with diversification at positions 1', 1, 2, 3, 4, and 6. Several synthetic challenges known to hinder the synthesis of this class of compounds were solved, including sensitivity to acids and bases, and epimerization and rearrangements, through the judicious choice of deprotection conditions, coupling conditions, and synthetic strategy. This work should enable a better understanding of structure-activity relationships in the streptogramin B compounds, possible identification of analogues that bypass known resistance mechanisms, and perhaps the identification of analogues with novel biological activities. PMID- 15352117 TI - A new phototransformation of methoxycarbonyl-substituted (E,Z,E)-1,3,5 hexatrienes: easy access to ring-annelated 8-oxabicyclo[3.2.1]octa-2,6-diene derivatives. AB - On attempting photochemically induced electrocyclizations of the previously reported 1,6-bismethoxycarbonyl- or 1,6-bistrimethylsilyl-substituted ring attached (E,Z,E)-1,3,5-hexatrienes 4 b, 4 c and 5 b, 5 c, equilibrium mixtures of the starting materials and their diastereomers, the corresponding (E,Z,Z) hexatrienes 4 b, 4 c and 5 b, 5 c were obtained. The desired trans-disubstituted ring-annelated cyclohexadienes 9 b and 10 b were formed by subsequent thermal 6pi electrocyclization of the (E,Z,Z)-hexatrienes 4 b and 5 b in good yields (77-83 %). Upon treating the bissilyl-substituted hexatrienes (E,Z,E)-5 c or (E,Z,Z)-5 c under the same conditions, 6pi-electrocyclizations did occur, but the primary products immediately isomerized to a large extent, and mixtures of the cyclohexane-annelated cyclohexadienes 10 c-12 c along with the dehydrogenation products 13, 14 c were formed. When the bismethoxycarbonyl-substituted hexatriene (E,Z,E)- 5 b was irradiated for an extended period of time (4.5 h), the gradual formation of the oxabicyclo[3.2.1]octa-2,6-diene 17 b by a formal intramolecular hetero-Diels-Alder reaction was observed and 17 b could be isolated in up to 69 % yield. To explore the scope of this new photochemical reaction, the new ring attached (E,Z,E)-hexatrienes 4 a, 5 a and 6 b were synthesized by twofold Heck reactions from 1,2-dibromocycloalkenes 1-3 (59-66 %). While irridiation of the cyclopentene-attached 1,3,5-hexatrienes only led to decomposition, the cyclohexene- and cycloheptene-attached hexatrienes gave the hetero-Diels-Alder products or other photoproducts depending on the size of the cycloalkene moiety and the nature of the alkoxycarbonyl substituents at the vinyl termini. The photoreaction products 17 b and 18 b which are bicyclic acetals, underwent cleavage upon treatment with a Lewis acid such as Me(3)SiOTf to give the ring annelated methoxycarbonyl-substituted troponecarboxylates 21 b and 22 b. PMID- 15352118 TI - A novel dimerization mode of a cyclic ketene imine. AB - The strained seven-membered cyclic ketene imine 9, obtained by cycloaddition of thiocarbonyl ylide 6 with 2,3-bis(trifluoromethyl)fumaronitrile (7), underwent base-catalyzed dimerization at room temperature on treatment with KCN in acetonitrile or with proton sponge in acetonitrile or CDCl(3). Two diastereoisomeric dimers, (6SR,3'RS)-13 and (6SR,3'SR)-13, were formed in 1:1 ratio in high yield. X-ray analysis revealed a deep-seated structural change which is unrelated to known dimerization pathways of ketene imines. In 13, one of the seven-membered rings is opened, and attached to the second unit by a thioimidate group. An ionic chain reaction with a formal fluoride ion as transfer agent offers a rationalization. PMID- 15352119 TI - Self-assembled metallocycles with two interactive binding domains. AB - Five metallocycles 1 a-e have been self-assembled from S-shaped bispyridyl ligands 2 a-e and a palladium complex, [Pd(dppp)(OTf)(2)] (dppp=1,3 bis(diphenylphosphanyl)propane), and have been characterized by elemental analysis and various spectroscopic methods including (1)H NMR spectroscopy and electrospray ionization (ESI) mass spectrometry. These metallocycles all are monocyclic compounds, but can fold to generate two binding domains bearing hydrogen-bonding sites based on pyridine-2,6-dicarboxamide units. The binding properties of the metallocycles with N,N,N',N'-tetramethylterephthalamide (G) have been probed by means of ESI mass spectrometry and (1)H NMR spectroscopy. The results both in the gas phase and in solution are consistent with the fact that the metallocycles accommodate two molecules of the guest G. Thus, the ESI mass spectra clearly show fragments corresponding to the 1:2 complexes in all cases. (1)H NMR studies on 1 a and G support the formation of a 1:2 complex in solution; the titration curves are nicely fitted to a 1:2 binding isotherm, but not to a 1:1 binding isotherm. In addition, a Job plot also suggests a 1:2 binding mode between 1 a and G, showing maximum complexation at approximately 0.33 mol fraction of the metallocycle 1 a in CDCl(3). The binding constants K(1) and K(2) are calculated to be 1600 and 1400 M(-1) (+/-10 %), respectively, at 25 degrees C in CDCl(3), indicative of positively cooperative binding. This positive cooperativity was confirmed by the Hill equation, affording a Hill coefficient of n = 1.6. Owing to insufficient solubility in CDCl(3), for comparison purposes the binding properties of the metallocycles 1 b-e were investigated in a more polar medium, 3 % CD(3)CN/CDCl(3). (1)H NMR titrations revealed that the metallocycles all bind two molecules of the guest G with Hill coefficients ranging from 1.4 to 1.8. This positive cooperativity may be attributed to a structural reorganization of the second binding cavity when the first guest binds to either one of the subcavities present in the metallocycles. PMID- 15352120 TI - The induced magnetic field in cyclic molecules. AB - The response of a molecule to an applied external magnetic field can be evaluated by a graphical representation of the induced magnetic field. We have applied this technique to four representative, cyclic organic molecules, that is, to aromatic (C(6)H(6), D(6h)), anti-aromatic (C(4)H(4), D(2h)) and non-aromatic (C(4)H(8), D(4h), and C(6)H(12), D(3d)) molecules. The results show that molecules that contain a pi system possess a long-range magnetic response, while the induced magnetic field is short-range for molecules without pi systems. The induced magnetic field of aromatic molecules shields the external field. In contrast, the anti-aromatic molecules increase the applied field inside the ring. Aromatic, anti-aromatic, and non-aromatic molecules can be characterized by the appearance of the magnetic response. We also show that the magnetic response is directly connected to nucleus-independent chemical shifts (NICS). PMID- 15352122 TI - Changes in the DNA methylation profile of the rat H19 gene upstream region during development and transgenic hepatocarcinogenesis and its role in the imprinted transcriptional regulation of the H19 gene. AB - Monoallelic expression of the imprinted H19 and insulin-like growth factor-2 (Igf2) genes depends on the hypomethylation of the maternal allele and hypermethylation of the paternal allele of the H19 upstream region. Previous studies from our laboratory on liver carcinogenesis in the F1 hybrid of Fischer 344 (F344) and Sprague-Dawley Alb SV40 T Ag transgenic rat (SD) strains revealed the biallelic expression of H19 in hepatomas. We undertook a comparative study of the DNA methylation status of the upstream region of H19 in fetal, adult, and neoplastic liver. Bisulfite DNA sequencing analysis of a 3.745-kb DNA segment extending from 2950 to 6695 bp of the H19 upstream region revealed marked variations in the methylation patterns in fetal, adult, and neoplastic liver. In the fetal liver, equal proportions of hyper- and hypomethylated strands revealed the differentially methylated status of the parental alleles, but in neoplastic liver a pronounced change in the pattern of methylation was observed with a distinct change to hypomethylation in the short segments between 2984 and 3301 bp, 6033-6123 bp, and 6518-6548 bp. These results indicated that methylation of all cytosines in this region may contribute to the imprinting status of the rat H19 gene. This phenomenon of differential methylation-related epigenetic alteration in the key cis-regulatory domains of the H19 promoter influences switching to biallelic expression in hepatocellular carcinogenesis. Similar to mouse and human, we showed that the zinc-finger CCTCC binding factor (CTCF) binds to the unmethylated CTCF binding site in the upstream region to influence monoallelic imprinted expression in fetal liver. CTCF does not appear to be rate limiting in fetal, normal, and neoplastic liver. 3' to the CTCF binding sites, another DNA region exhibits methylation of CpG's in both DNA strands in adult liver, retention of the imprint in fetal liver, and complete demethylation in neoplastic liver. In this region is also a putative binding site for a basic helix-loop-helix leucine-zipper transcription factor, TFEB. The differential CpG methylation seen in the adult that involves the TFEB binding site may explain the lack of expression of the H19 gene in adult normal liver. Furthermore, these findings demonstrate that the loss of imprinting of the H19 gene in hepatic neoplasms of the SD Alb SV40 T Ag transgenic rat is directly correlated with and probably the result of differential methylation of CpG dinucleotides in two distinct regions of the gene that are within 4 kb 5' of the transcription start site. Cytogenetic analysis of hepatocytes in the transgenic animal prior to the appearance of nodules or neoplasms indicates a role of such loss of imprinting in the very early period of neoplastic development, possibly the transition from the stage of promotion to that of progression. PMID- 15352123 TI - Gene expression microarray analysis and genome databases facilitate the characterization of a chromosome 22 derived homogeneously staining region. AB - Karyotype and fluorescence in situ hybridization (FISH) analyses previously identified a homogeneously staining region (HSR) derived from chromosome 22 in OV90, an epithelial ovarian cancer (EOC) cell line. Affymetrix expression microarrays in combination with the UniGene and Human Genome Browser databases were used to identify the candidate genes comprising the amplicon of the HSR, based on comparison of expression profiles of OV90, EOC cell lines lacking HSRs and primary cultures of normal ovarian surface epithelial (NOSE) cells. A group of probe sets displaying a minimum 3-fold overexpression with a high reliability score (P-call) in OV90 were identified which represented genes that mapped within a 1-2 Mb interval on chromosome 22. A large number of probe sets, some of which represent the same genes, displayed no evidence of overexpression and/or low reliability scores (A-call). An investigation of the probe set sequences with the Affymetrix and Sanger Institute Chromosome 22 Group databases revealed that some of the probe sets displaying discordant results for the same gene were complementary to intronic sequences and/or the antisense strand. Microarray results were validated by RT-PCR. Genomic analysis suggests that the HSR was derived from the amplification of a 1.1 Mb interval defined by the chromosomal map positions of ZNF74 and Hs.372662, at 22q11.21. The deduced amplicon is derived from a complex region of chromosome 22 that harbors low-copy repeats (LCRs). The amplicon contains 18 genes as likely targets for gene amplification. This study illustrates that large-scale expression microarray analysis in combination with genome databases is sufficient for deducing target genes associated with amplicons and stresses the importance of investigating probe set design before engaging in validation studies. PMID- 15352124 TI - Proto oncogene/eukaryotic translation initiation factor (eIF) 4E attenuates mevalonate-mediated regulation of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase synthesis. AB - The rate-limiting enzyme for mevalonate synthesis in mammalian cells is 3-hydroxy 3-methylglutaryl coenzyme A (HMG-CoA) reductase. Products of mevalonate synthesis are required for cell cycle progression as well as cell growth and survival. In tumor cells, HMG-CoA reductase is generally elevated because of attenuated sterol mediated regulation of transcription. However, tumor cell HMG-CoA reductase remains sensitive to post-transcriptional regulation by mevalonate-derived isoprenoid intermediates of cholesterol synthesis. Isoprenoids suppress HMG-CoA reductase synthesis through a mechanism that reduces initiation of translation on HMG-CoA reductase mRNA. Because HMG-CoA reductase mRNA transcripts have 5' untranslated regions (UTR) that are GC rich and contain stable secondary structure, we tested the hypothesis that overexpression of eIF4E would attenuate isoprenoid-mediated regulation of HMG-CoA reductase. eIF4E is elevated in many tumor cells and behaves as a proto-oncogene by aberrantly translating mRNAs whose translation is normally suppressed by 5-UTRs that are GC rich. A CHO cell line expressing high levels of eIF4E (rb4E) was developed by infecting cells with retroviruses containing a full-length mouse cDNA for eIF4E. Levels of reductase synthesis were elevated fivefold in rb4E cells compared to noninfected CHO cells; HMG-CoA reductase mRNA levels were not increased in rb4E cells compared to normal CHO cells. Total cellular protein synthesis was only increased by approximately 15% in rb4E cells compared to CHO cells. The mTOR inhibitor rapamycin lowered HMG CoA reductase synthesis by 50 and 60% in rb4E and CHO cells, respectively; no equivalent effect was observed for HMG-CoA reductase mRNA levels with rapamycin treatment. These results indicate that HMG-CoA reductase mRNA is in a class of mRNAs with highly structured 5'-UTRs whose m(7)GpppX cap-dependent translation is closely linked to the rapamycin-sensitive mitogen activated pathway for protein synthesis. PMID- 15352125 TI - Increased DNA methylation in the HoxA5 promoter region correlates with decreased expression of the gene during tumor promotion. AB - Promoter-region DNA methylation inhibits transcription. A two-stage SENCAR (sensitive to mouse carcinogenesis) mouse skin carcinogenicity model was used to examine gene-specific changes in methylation during skin tumor promotion. Analysis was performed on 7,12-dimethylbenz[a]anthracene (DMBA)-initiated skin promoted with 9, 18, 27, or 36 mg cigarette smoke condensate (CSC) for 9 wk, or 27 mg CSC for 9 wk and sacrificed 6 wk afterwards (recovery group). Additionally, tumors arising following promotion with 27 mg CSC for 29 wk were assessed. Gene array analysis identified differentially expressed genes. Expression of HoxA5, a tumor suppressor gene, was decreased following 9 wk of treatment with 27 mg CSC, and returned to control levels during recovery. HoxA5 promoter methylation was measured with the enzymatic regional methylation assay (ERMA). DNA was bisulfite modified, PCR-amplified with primers containing dam sites, incubated with [14C methyl] S-adenosyl-L-methionine (SAM) and dam methyltransferase for DNA quantification, then incubated with [3H-methyl] SAM and SssI methylase to quantify methylation status. Higher 3H/14C ratios indicate increased methylation. The 3H/14C ratios of animals promoted with 27 or 36 mg CSC (48.2 +/- 6.9 and 24.2 +/- 6.1, respectively) were higher than the control or recovery group ratios (12.3 +/- 0.1 and 12.6 +/- 0.3, respectively); sequence analysis supported these findings. Increased methylation of p16 or O6 methylguanine methyltranferase (MGMT) was detected in 4/8 (50%) of the tumor samples from mice promoted with 27 mg CSC. These data suggest that increased DNA methylation contributes to the downregulation of HoxA5, and combined with hypermethylation of p16 or MGMT, this might facilitate the clonal expansion of increasingly aberrant cells during promotion. PMID- 15352127 TI - Environmental Mutagen Society 35th Annual Meeting, October 2-6, 2004, Pittsburgh, Pennsylvania, USA. Abstracts. PMID- 15352131 TI - The hedonic response to cigarette smoking is proportional to dopamine release in the human striatum as measured by positron emission tomography and [11C]raclopride. AB - Positron emission tomography and [11C]raclopride were used to assess the dopaminergic response to cigarette smoking in ten smokers. Nicotine-deprived smokers were scanned twice on separate days. In one condition, participants smoked their usual brand of cigarettes while in the scanner and in the other condition they remained nicotine abstinent. On each day, subjects monitored the hedonic properties of their experience as well as their levels of craving. Initial analyses revealed no significant differences between the conditions in [11C]raclopride binding potential (BP) in the caudate, putamen, or ventral striatum. Because previous research suggested that drug-induced dopamine transmission is related to levels of craving and/or hedonic drug effects, the relationship between these variables and [11C]raclopride BP was examined. Craving levels were reduced by smoking but were not systematically related to BP change. However, the hedonic response to smoking was correlated with BP reduction in the caudate (P < 0.001) and posterior putamen (P < 0.05) but not in the ventral striatum. Post hoc analyses revealed that only five of the ten smokers reported mood-elevating effects in response to the smoking procedure. In these subjects, smoking was associated with decreased [11C]raclopride BP in the caudate. On the other hand, among subjects that reported a diminished mood response to smoking there was an increase in BP in the caudate and putamen. These results suggest that pleasurable drug experiences are associated with increased dopamine transmission in the dorsal striatum while unpleasant experiences may be related to decreased dopamine release in this region. PMID- 15352132 TI - Altered emotional behaviors and the expression of 5-HT1A and M1 muscarinic receptors in micro-opioid receptor knockout mice. AB - Anxiety and depression alterations have been reported in micro-opioid receptor knockout mice after exon 2 disruption. However, emotional behaviors, such as novelty and emergence responses have not been reported in micro-opioid receptor knockout mice due to the disruptions of exon 2 and 3. Here, we report that mu opioid receptor knockout mice, with deletion of exon 2 and 3, display significant emotional behavior changes; they showed less anxiety in the elevated plus maze and emergence tests, reduced response to novel stimuli in the novelty test, and less depressive-like behavior in the forced-swim test. Analysis of the compensatory mechanism in mu-opioid receptor knockout mice revealed that the M1 mRNA levels were reduced in the cortex, caudate putamen, nucleus accumbens, and hippocampus, and that M1 receptor levels were reduced in the nucleus accumbens, CA1, and the dentate gyrus of the hippocampus, versus the wild-type. However, 5 HT1A receptor levels were significantly elevated in the cerebral cortex and in the hypothalamus of mu-opioid receptor knockout mice versus the wild-type. These aberrant emotional behavioral phenotypes are possibly related to M1 and 5-HT1A receptor alterations in the micro-opioid receptor knockout mice. Overall, our study suggests that micro-opioid receptor may play a role in the modification of emotional responses to novelty, anxiety, and depression. PMID- 15352133 TI - Binding characteristics of high-affinity dopamine D2/D3 receptor agonists, 11C PPHT and 11C-ZYY-339 in rodents and imaging in non-human primates by PET. AB - We have evaluated the in vitro autoradiographic binding characteristics and in vivo brain distribution of two high-affinity dopamine D2/D3 receptor agonists, (+/-)-2-(N-phenethyl-N-1'-11C-propyl)amino-5-hydroxytetralin (11C-PPHT) and (+/-) 2-(N-cyclohexylethyl-N-1'-11C-propyl)amino-5-hydroxytetralin (11C-ZYY-339) in rodents and in monkeys using positron emission tomography (PET). In vitro autoradiograms in rat brain slices with (11)C-PPHT and 11C-ZYY-339 revealed binding to dopaminergic regions in the striata, which was substantially (>90%) displaced by 10 microM sulpiride. Striatal binding was also removed in the presence of 5-guanylylimidophosphate (Gpp(NH)p), indicative of binding of these radiotracers to the high-affinity (HA) state. The results of in vivo studies in rats exhibited binding of the two radiotracers to the striata (striata/cerebellum approached 2 in 30 min). The regional selectivity to the striata was reduced by preadministration of haloperidol. PET studies in male rhesus monkeys using an ECAT EXACT HR+ scanner indicated localization of 11C-PPHT and 11C-ZYY-339 in the striata and thalamus. Striata to cerebellum and thalamus to cerebellum ratios were low (1.5 and 1.3, respectively, at 30 min postinjection) for both 11C-PPHT and 11C-ZYY-339, apparently due to the slower nonspecific clearance from cerebellum. These findings with 11C-PPHT and 11C-ZYY-339 indicate the possibility of in vivo imaging of high-affinity state of dopamine D2/D3 receptors in both the striata and the thalamus. PMID- 15352134 TI - Long-term prenatal hypoxia alters maturation of brain catecholaminergic systems and motor behavior in rats. AB - In order to determine the influence of long-term prenatal hypoxia on the maturation of the brain catecholaminergic structures involved in motor and cognitive functions, pregnant rats were subjected to hypoxia (10% O2) from the 5th to 20th day of gestation. The in vivo activity of tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis, was assessed, by accumulation of L-DOPA after i.p. administration of NSD-1015, in the motor cortex areas, the hippocampus, and the striatum at birth and at the 3rd, 7th, 14th, 21st, and 68th postnatal days. The motor reactivity to novelty and the circadian motor activity were measured at the 21st and 68th postnatal days. Exposure to prenatal hypoxia strongly altered the developmental pattern of in vivo TH activity in restricted noradrenergic terminals of the brain. In the 21-day-old prenatal hypoxic rats, the TH activity was reduced by 80% in the motor cortex areas and by 43% in the hippocampus, compared to control rats, while no differences could be detected in the striatum. Compared to control rats, the prenatal hypoxic pups exhibited a higher motor reactivity to novelty and a nocturnal motor hypoactivity at the 21st postnatal day. The neurochemical and behavioral alterations were no longer observed at the 68th postnatal day. The altered in vivo TH activity in the young rats might be part of the neural mechanisms contributing to the motor behavioral impairments induced by prenatal hypoxia. Long-term prenatal hypoxia could be linked to the development of psychopathologies that can be detected in infancy. PMID- 15352135 TI - Modulation of bradykinin-induced calcium changes in SH-SY5Y cells by neurosteroids and sigma receptor ligands via a shared mechanism. AB - In this study we investigated the effects of sigma receptor ligands and neurosteroids on bradykinin-induced intracellular calcium concentration ([Ca2+]i) changes in SH-SY5Y neuroblastoma cells. [Ca2+]i levels in cells loaded with fura 2 were monitored with dual-wavelength ratiometric fluorescence measurement. Submicromolar concentrations of bradykinin elicited [Ca2+]i responses with a fast rise followed by a slow decline in these cells. Preincubation of low micromolar concentrations of the neurosteroids pregnenolone, dehydroepiandrosterone (DHEA), or the prototypic sigma (sigma) receptor agonist (+)pentazocine potentiated bradykinin-induced [Ca2+]i changes in SH-SY5Y cells. The sigma receptor antagonist haloperidol blocked the enhancing effects on [Ca2+]i by (+)pentazocine or pregnenolone. Progesterone did not significantly affect the basal [Ca2+]i level or bradykinin-induced [Ca2+]i changes in these cells. However, coincubation of progesterone with (+)pentazocine, pregnenolone, or DHEA reversed their potentiating effects. The antagonistic effects of haloperidol and progesterone on the potentiating effects of (+)pentazocine and pregnenolone suggested that these ligands might act through a common mechanism. We further showed that progesterone, pregnenolone, and DHEA competed for [3H]+pentazocine binding in SH SY5Y cells with Ki values of 0.13 +/- 0.03 microM, 0.98 +/- 0.34 microM, and 5.2 +/- 1.4 microM, respectively. Thus, the modulation of bradykinin-induced [Ca2+]i changes by neurosteroids in these cells is likely due to their actions on sigma receptors. PMID- 15352136 TI - Serotonin-induced phase advances of SCN neuronal firing in vitro: a possible role for 5-HT5A receptors? AB - Spontaneous firing rates of neurons in the suprachiasmatic nuclei (SCN) follow a consistent pattern, peaking near the midpoint of the light phase in a 12:12 light/dark schedule, and repeating this brief period of increased activity in subsequent circadian cycles. These carefully timed fluctuations reflect the output signal of the SCN, long recognized as the site of the endogenous biological clock in mammals. In rat hypothalamic slices, bath incubations of 8-OH DPAT had previously been shown to elicit phase advances when applied at ZT6 (or 6 h following the onset of light), an action that could readily be attributed to 5 HT7 receptor activation. The present studies set out with the simple goal of establishing that the same receptor mechanism was responsible for the phase shifting actions of 5-HT itself. Surprisingly, the phase advances elicited by 5 HT (0.5 microM, 1 h) at ZT6 were reduced by one 5-HT7 antagonist, ritanserin (10 microM), but not by another, mesulergine (10 microM). Receptor binding studies demonstrated a 25-fold greater affinity of ritanserin for h5-HT5A sites compared to mesulergine (Ki = 71 nM vs. 1,800 nM), an observation suggestive of a 5-HT5A mechanism for 5-HT and consistent with earlier observations of robust labeling of 5-HT5A sites in the SCN. 5-HT generated by the addition of L-tryptophan (10 microM, 1 h) to the slices displayed the same pattern of sensitivity, that is, blockade by ritanserin but not by mesulergine. Rp-cAMPS, a cAMP antagonist, failed to block the phase shifts elicited by 5-HT at a concentration (1 microM) previously shown to be effective against 8-OH-DPAT-induced phase shifts, in keeping with the proposed negative coupling of 5-HT5A receptors to cAMP production. Taken together, these results suggest that activation of both 5-HT5A and 5-HT7 receptors can produce phase advances of the circadian clock in vitro when they occur during mid-subjective day. PMID- 15352137 TI - Subthalamic nucleus lesions alter basal and dopamine agonist stimulated electrophysiological output from the rat basal ganglia. AB - The subthalamic nucleus (STN) is an important link in the "indirect" striatal efferent pathway. To assess its role on basal ganglia output via the substantia nigra pars reticulata (SNr), we monitored the single unit activities of SNr neurons in chloral hydrate-anesthetized rats 5-8 days after bilateral kainic acid lesions (0.75 microg/0.3 microl/side) of the STN. Consistent with loss of an excitatory input, the average basal firing rate of SNr neurons was significantly reduced in STN-lesioned animals. Moreover, the lesions modified the responses of SNr neurons to individual and concurrent stimulation of striatal D1 and D2 receptors. Bilateral striatal infusions of the D1/D2 agonist apomorphine (10 microg/microl/side) into the ventral-lateral striatum (VLS) were previously shown to cause significant increases in SNr cell firing (to 133% of baseline) in normal rats. However, in STN-lesioned rats, identical infusions caused no overall change in SNr activity (mean, 103% of basal rates). Conversely, selective stimulation of striatal D2 receptors by bilateral co-infusion of the D2 agonist quinpirole and the D1 antagonist SCH 23390 that previously caused little change in SNr firing in normal rats significantly inhibited their firing in STN-lesioned rats. Finally, the modest excitatory responses of SNr neurons to selective stimulation of striatal D1 receptors by co-infusions of SKF 82958 with the D2 antagonist YM09151 2 were not altered by lesions of the STN. These results implicate the STN as a mediator of excitatory response of SNr neurons to D2, and mixed D1/D2, dopamine receptor agonists in normal rats, and challenge conventional views on the role of the STN and the "indirect" pathway in regulating dopamine-stimulated output from the SNr. PMID- 15352138 TI - A longitudinal study of Alzheimer's disease: rates of cognitive and functional decline. AB - OBJECTIVE: To measure rates of decline in cognition and function in patients with Alzheimer's disease (AD) and to investigate their accelerating risk factors in Korea. METHODS: This study presents longitudinal data on a community-based sample of 107 patients with AD, followed at 6 months and 12 months. The cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-cog), the Mini Mental State Examination (MMSE) and the Disability Assessment for Dementia Scale (DAD) were given. Mixed model analyses were conducted using the following independent variables: times of repeated assessment (0, 6 or 12 months), severity of dementia assessed by the Functional Assessment Staging (FAST) and individual indicators as covariates. RESULTS: Average annual rates of decline in the MMSE, the ADAS-cog and the DAD were 2.3, 11.4 and 15.1 points, respectively. Neither gender, duration of formal education, nor duration of AD since onset was significant predictors of cognitive and functional decline. Patterns of functional decline in total DAD, instrumental ADLs, planning and organization and performance subscale are linear as MMSE score declines, while those of the basic ADLs and the initiation are curvilinear. CONCLUSION: This naturalistic observational study measured rates of cognitive and functional decline in AD, and can provide reference data for further longitudinal studies or clinical trials. Further study will be necessary to determine whether linear or curvilinear pattern in functional decline is due to progression of AD itself or statistical artifact. PMID- 15352139 TI - Combined effect of vision and hearing impairment on depression in elderly Chinese. AB - BACKGROUND: Sensory impairment and depression are common in old age and the relation between depression and vision as well as hearing impairment have been established. However, few studies have directly compared their effects and examined the impact of dual sensory loss. The purpose of this study is to compare impacts of self-reported hearing and vision loss as well as the effect of double sensory impairment on depression. METHOD: This article analyzes cross-sectional data collected from a representative community sample of 2,003 Chinese elderly people aged 60 or above in Hong Kong. Respondents were interviewed in a face-to face format and data including vision and hearing impairment, socio-demographic variables, health indicators, family support, and depression were obtained. RESULTS: Logistic regression analyses revealed that visual impairment was significantly related to depression even after age, gender, marital status, education, self-reported health status, the presence of 11 diseases, functional limitation and family support were controlled but hearing loss was not. Hearing impairment did not add to the likelihood of depression where visual impairment was already present. CONCLUSION: The impact of visual impairment on psychological well-being among elderly Chinese is more robust than hearing loss. Therefore, aged care service practitioners must take this risk factor into consideration in their preventive intervention and treatment for psychological well-being. PMID- 15352140 TI - Antidepressant efficacy and cognitive effects of repetitive transcranial magnetic stimulation in vascular depression: an open trial. AB - BACKGROUND: Beneficial effects of repetitive transcranial magnetic stimulation (rTMS) were demonstrated by many controlled studies in major depression. Moreover, this promising and non invasive therapeutic tool seems to be better tolerated than electroconvulsive therapy.Vascular depression is a subtype of late life depression, associated with cerebrovascular disease and means a poorer response to antidepressant treatment. We employed rTMS over the left prefrontal cortex in 11 patients with late-onset resistant vascular depression. The primary purpose of this two-week open study was to examine antidepressant efficacy of rTMS in vascular depression. The secondary aim was to evaluate cognitive effects of rTMS in our sample. METHODS: Clinical status, as measured with the Hamilton Depression Rating Scale (HDRS), and cognitive effects, as evaluated by neuropsychological tests, were assessed at baseline and after two weeks of rTMS. Brain measurements to obtain an index of prefrontal atrophy were performed at both the motor cortex and prefrontal cortex. RESULTS: Five out of 11 resistant patients with late-onset vascular depression were responders. They showed a clinically meaningful improvement in HDRS scores, with a decrease of 11, 4 points (p<0.01). Antidepressant response is correlated to the relative degree of prefrontal atrophy (p = 0.05). After two weeks, verbal fluency and visuospatial memory improved. No cognitive performance deteriorated except for verbal memory, as the delayed recall decreased significantly in the responders' group. CONCLUSIONS: Our preliminary observations prompt to perform a subsequent controlled study to examine if rTMS may constitute an alternative to electroconvulsive therapy. PMID- 15352141 TI - Lifetime urban/rural residence, social support and late-life depression in Korea. AB - BACKGROUND: Population ageing and rural-urban migration are accelerating in many non-Western nations. This study aimed to investigate: (i) the association between lifetime urban/rural residence and late-life depression in Korea and (ii) modification of associations between depression and social support by lifetime residence. METHODS: 1204 urban/rural residents aged 65+were interviewed and GMS AGECAT diagnoses made. Previous areas of residence were recorded and social support deficits quantified. RESULTS: Depression was present in 9% and 21% of the rural and urban samples respectively. For the urban sample, depression was not associated with earlier urban/rural residence. Social support deficits were most strongly associated with depression in people with a lifetime rural residence, followed by urban residents with a rural birthplace. CONCLUSIONS: Prevalence rates of depression were increased in the urban sample regardless of previous urban/rural residence. Reduced social support was particularly strongly associated with depression in people with a rural upbringing. PMID- 15352142 TI - Does depression in old age increase only cardiovascular mortality? The Leiden 85 plus Study. AB - BACKGROUND: Depression in old age is associated with an increased mortality risk of cardiovascular disease but the mortality risk from non-cardiovascular causes is disputed. OBJECTIVE: To investigate the effect of depression on cardiovascular and non-cardiovascular mortality in old age. METHODS: We prospectively followed 500 subjects from age 85 years onwards within the population-based Leiden 85-plus Study. Depressive symptoms were assessed annually with the 15-item Geriatric Depression Scale (GDS-15). Mortality risks were estimated in a Cox proportional hazards model with the annual assessment of depression (GDS-15> or =4 points) as a time-dependent covariate. RESULTS: During 1654 person-years of follow-up (mean per person, 3.2 years), depression was associated with a two-fold increase of all cause mortality [Relative Risk (RR), 1.83; 95% Confidence Interval (CI), 1.24 2.69] that was not explained by comorbid conditions. Both cardiovascular mortality and non-cardiovascular mortality contributed equally to the excess mortality (RR 1.95 and 1.75 respectively). CONCLUSION: Depression in old age contributes to an increase of both cardiovascular and non-cardiovascular mortality. Motivational depletion may play an important role in the increased mortality in elderly with depression. PMID- 15352143 TI - A randomized controlled trial of a behavior advisory service for hospitalized older patients with confusion. AB - OBJECTIVE: To determine whether individualized advice on non-pharmacological strategies for hospitalized older patients with confusion and behavioral problems can improve levels of agitation and reduce the use of psychotropic medication. METHOD: Pragmatic randomized controlled trial in two metropolitan teaching hospitals in South Australia. Seventy-one older patients with confusion and a behavioral disturbance were randomly allocated to receive either an assessment and ongoing individualized advice from an extended practice nurse on non pharmacological strategies or usual care. Usual care included an assessment by a geriatrician. MEASUREMENTS: Primary outcomes were levels of agitation (Pittsburgh Agitation Scale), appropriateness of psychotropic medication prescribing (Medication Appropriateness Index) and levels of psychotropics administered (chlorpromazine and diazepam dose equivalents). Secondary outcomes were length of stay, discharge destination, number of falls, restraint use and satisfaction from nursing staff and next of kin. RESULTS: Levels of agitation were modest at baseline with mean PAS intervention group 3.4 (SD = 0.5) and control group 4.0 (SD = 0.4) and both groups improved over time to 1.7 (SD = 0.4) for the intervention group and 1.8 (SD = 0.3) for the control group on the final day of data collection. Median length of follow-up was nine days. There was no effect of the intervention on levels of agitation, amount and appropriateness of psychotropic medication prescribed and administered, falls, length of stay, discharge destination, restraint use and nursing and next of kin satisfaction. CONCLUSION: A nursing consultation service providing individualized non pharmacological advice does not improve patient agitation or use of psychotropic medication for older patients with confusion and behavioral problems in an acute hospital. PMID- 15352144 TI - Brain perfusion correlates of the apathy inventory dimensions of Alzheimer's disease. AB - BACKGROUND: Apathy is defined as a lack of motivation in behavior, cognition and affect. This syndrome is frequent in various neuropsychiatric diseases but little is known about its pathophysiology. OBJECTIVES: The aim of this study was to investigate the metabolic correlates of the behavioral, cognitive and emotional, aspects of apathy in Alzheimer's disease (AD). METHOD: Thirty AD patients were included. Lack of initiative, lack of interest and of emotional blunting were assessed with the Apathy Inventory (IA), a tool designed to provide a separate assessment of the behavioral, cognitive and emotional, aspects of apathy. Brain perfusion was measured by (99m)Tc-labeled bicisate (ECD) single photon emission tomography. RESULTS: The Statistical Parametric Mapping software provides negative correlation between IA total score and brain perfusion in left and right superior orbito-frontal gyrus, and to a lesser extent in left middle frontal gyrus (BA10). Lack of initiative score was negatively correlated with perfusion in right anterior cingulate cortex. Lack of interest score was negatively correlated with perfusion in right middle orbitofrontal gyrus). Emotional blunting score correlated negatively with in left superior dorsolateral prefrontal cortex activity. CONCLUSION: These results underline that the cognitive, behavioral and affective components of motivation are mediated by different fronto-sub-cortical circuits and are differently lateralized. In particular, left prefrontal hypoperfusion is involved in emotional blunting, as it was often demonstrated in depressive disorders. These distinct components of apathy may be targeted by different therapeutic means, in which dopaminergic enhancement might play a major role. PMID- 15352145 TI - Characteristics of the GPCOG, a screening tool for cognitive impairment. AB - BACKGROUND: Early dementia diagnosis is aided by the use of brief screening tests; scores can be biased by patient and informant characteristics such as age, gender and education. OBJECTIVE: To assess whether the General Practitioner's Assessment of Cognition (GPCOG), a brief screening tool for detecting cognitive impairment comprising a patient cognitive test and questions to an informant, is biased by patient and informant characteristics. DESIGN: Sixty-seven general practitioners recruited consecutive patients (with informants). Patients were subsequently assessed by a research psychologist, and DSM-IV diagnoses assigned following a case-conference. SETTING: Primary Care. SUBJECTS: Two hundred and eighty three home-dwelling individuals, 11.3% of whom were aged 50-74 years with suspected memory problems and the rest aged 75 or more. METHODS: The GPCOG, Cambridge Mental Disorder of the Elderly Examination cognitive scale (CAMCOG), Geriatric Depression Scale (GDS), and the SF-12 Health Survey (SF-12) were administered and demographic data were collected and consensus DSM-IV diagnoses of dementia made. Relationships between patient and informant characteristics and the GPCOG measure were examined using Pearson correlations and linear regression analyses. RESULTS: There were correlations in GPCOG-patient scores with age, education and depression scores but on regression analysis only age was associated with the GPCOG-patient section. The GPCOG-informant section was free of bias. CONCLUSIONS: The GPCOG has advantages for use in primary care and is free of many biases common in other scales. PMID- 15352146 TI - Telephone word-list recall tested in the rural aging and memory study: two parallel versions for the TICS-M. AB - BACKGROUND: Parallel versions of memory tasks are useful in clinical and research settings to reduce practice effects engendered by multiple administrations. We aimed to investigate the usefulness of three parallel versions of ten-item word list recall tasks administered by telephone. METHODS: A population based telephone survey of middle-aged and elderly residents of Bradley County, Arkansas was carried out as part of the Rural Aging and Memory Study (RAMS). Participants in the study were 1845 persons aged 40 to 95 years. Word lists included that used in the telephone interview of cognitive status (TICS) as a criterion standard and two newly developed lists. RESULTS: The mean age of participants was 61.05 (SD 12.44) years; 39.5% were over age 65. 78% of the participants had completed high school, 66% were women and 21% were African-American. There was no difference in demographic characteristics between groups receiving different word list versions, and performances on the three versions were equivalent for both immediate (mean 4.22, SD 1.53) and delayed (mean 2.35 SD 1.75) recall trials. The total memory score (immediate+delayed recall) was negatively associated with older age (beta = -0.41, 95%CI=-0.11 to -0.04), lower education (beta = 0.24, 95%CI = 0.36 to 0.51), male gender (beta = -0.18, 95%CI = -1.39 to -0.90) and African-American race (beta = -0.15, 95%CI = -1.41 to -0.82). CONCLUSIONS: The two RAMS word recall lists and the TICS word recall list can be used interchangeably in telephone assessment of memory of middle-aged and elderly persons. This finding is important for future studies where parallel versions of a word-list memory task are needed. (250 words). PMID- 15352147 TI - Assessment of agitation in elderly patients with dementia: correlations between informant rating and direct observation. AB - BACKGROUND: Assessment of behavior problems in elderly persons with dementia is important for understanding and managing those behaviors. The most common method for assessing agitation is the use of informant ratings; however, these ratings may be affected by staff bias, inaccurate or insufficient memory, or stress. An alternative method is direct observation, which is more objective, but very costly and necessitates time sampling, thereby limiting the period covered by the assessment. To date, little research attention has been given to the degree to which these two methods converge. METHODS: In the present study, 175 elderly persons with dementia who manifested problem behaviors were recruited from 11 nursing home facilities in Maryland. The average age for the participants was 87 years; 78% were female. Two methods were employed for assessing agitation: the Agitated Behaviors Mapping Instrument (ABMI), which is based upon direct observations, and the Cohen-Mansfield Agitation Inventory (CMAI), which is a frequency rating scale completed by a formal caregiver. The ABMI and CMAI contain some identical items for tapping behavior problems. RESULTS: Data analysis revealed significant Pearson correlations between identical items on the two assessment instruments, as well as significant correlations of summary measures based on these different instruments, demonstrating a strong convergence between informant ratings and direct observations. CONCLUSIONS: Informant ratings can achieve moderate agreement with direct observation when valid instruments and informants are used. PMID- 15352148 TI - Effectiveness of a clinical pathway for the diagnosis and treatment of dementia and for the education of families. AB - AIMS: Clinical pathways (CPs) are rarely used in the treatment of dementia. We established a CP for a series of medical practices (diagnosis, treatment, establishment of a care system, and caregiver education) for patients with dementia hospitalized for a three-week period, and evaluated its usefulness. METHODS: The length of hospital stay and hospital costs were compared between 23 consecutive patients with dementia hospitalized and treated using a CP and 20 controls treated by conventional medical practice without using a CP in a special ward for dementia patients. In the CP group, at the time of discharge, primary caregivers, physicians, and nurses were given a questionnaire to obtain their comments about the impression of treatment with the CP. RESULTS: The questionnaire survey indicated that the CP deepened the caregiver's understanding of the sequence of medical practices for the inpatient, the disorders of the inpatient, the treatment methods, and the methods for coping with the disorder. The CP was also useful for facilitating inpatient medical practice and promoting the establishment of a care system after discharge. The use of the CP significantly shortened the length of hospital stay and decreased hospital costs during hospitalization but increased the amount of work per day and made the medical staff feel that their freedom to choose medical procedures had been restricted. CONCLUSIONS: The CP was useful for execution of inpatient medical practices for patients with dementia. PMID- 15352149 TI - Efficacy and safety of long-acting risperidone in elderly patients with schizophrenia and schizoaffective disorder. AB - BACKGROUND: Elderly patients are often an underserved population in terms of optimizing treatment outcomes. Long-acting risperidone, the first long-acting injectable atypical antipsychotic, can improve outcomes through continuous medication delivery. OBJECTIVE: To assess the efficacy and safety of long-acting injectable risperidone in elderly patients with psychotic disorders. METHODS: This is a subanalysis of 57 patients aged > or =65 years enrolled in an open label study of long-acting risperidone that included 725 symptomatically stable patients with schizophrenia or schizoaffective disorder. Patients were assigned to receive 25, 50, or 75 mg of long-acting risperidone every 2 weeks for up to 50 weeks. RESULTS: Fifty-seven elderly patients (mean +/- SE age, 70.9 +/- 0.7 years) were enrolled. Mean Positive and Negative Syndrome Scale (PANSS) total scores improved significantly throughout the study and at endpoint (p < 0.001). The PANSS factor scores (positive symptoms, negative symptoms, disorganized thoughts, uncontrolled hostility/excitement, and anxiety/depression) also significantly improved (p < 0.01). Clinical improvement (> or =20% reduction in PANSS total scores) was achieved by 49% of these stable patients, and 55% improved on the Clinical Global Impressions scale. Severity of movement disorders (Extrapyramidal Symptom Rating Scale scores) was reduced significantly. Adverse events reported in >10% of patients were insomnia (14%), constipation (12%), and bronchitis (12%). CONCLUSIONS: Long-acting risperidone was associated with significant symptom improvements in stable elderly patients with schizophrenia or schizoaffective disorder. Treatment was well tolerated. PMID- 15352150 TI - Persistent Deja vu: a disorder of memory. PMID- 15352151 TI - Characterization of triacylglycerols from overwintering prepupae of the alfalfa pollinator Megachile rotundata (Hymenoptera: Megachilidae). AB - Alfalfa leafcutting bees, Megachile rotundata (F.), overwinter as prepupae. The internal lipids were extracted from prepupae that had been wintered at 4 degrees C for 7 months. Megachile rotundata prepupae possessed copious quantities of internal lipids (20% of the fresh weight) that were extracted with CHCl3/methanol (2:1). Transmission electron microscopy revealed that lipids were stored within very large intracellular vacuoles. Separation by silica chromatography revealed that 88% of the internal lipids were triacylglycerols. Ester derivatives of fatty acids from triacylglycerol components were analyzed by gas chromatography-mass spectrometry and 15 fatty acid constituents were identified. The majority (76%) of the triacylglycerol fatty acids were unsaturated fatty acids. The major triacylglycerol fatty acid constituent (30%) was the C16 monounsaturated fatty acid, palmitoleic acid (16:1, hexadec-9-enoic acid), with substantial amounts of linolenic acid (18:3, octadec-9,12,15-trienoic acid, 15%), palmitic acid (16:0, hexadecanoic acid, 14%) and oleic acid (18:1, octadec-9-enoic acid, 13%). Palmitoleic acid as the major fatty acid of an insect is an unusual occurrence as well as the presence of the 16-carbon polyunsaturated fatty acids, 16:2 and 16:3. The major intact triacylglycerol components were separated and identified by high performance liquid chromatography-mass spectrometry. A complex mixture of approximately 40 triacylglycerol components were identified and major components included palmitoyl palmitoleoyl oleoyl glycerol, palmitoyl palmitoleoyl palmitoleoyl glycerol, myristoyl palmitoleoyl palmitoleoyl glycerol, myristoleoyl palmitoyl palmitoleoyl glycerol, and palmitoyl palmitoleoyl linolenoyl glycerol. The function of these internal lipids and their relevance to winter survival and post-wintering development of M. rotundata is discussed. PMID- 15352152 TI - Molecular cloning and bacterial expression of pheromone binding protein in the antennae of Helicoverpa armigera (Hubner). AB - A cDNA clone coding for pheromone binding protein was isolated from the antennae of Helicoverpa armigera by RT-PCR and (5'/3')-RACE technique. The full-length of H. armigera pheromone binding protein (HarmPBP) was 952 bp, possessing 162 amino acid residues including a signal peptide of 20 amino acids. Its predicted molecular weight and isoelectric point were 18.26 kDa and 5.23, respectively. This deduced amino acid sequence shared some common structural features with odorant-binding proteins from several moth species, including the six conserved cysteine motif, a typical characteristic of insect's odorant-binding proteins. Northern blot showed that HarmPBP is specifically expressed in the antennae of Helicoverpa armigera and more abundantly expressed in male than female. During the antennal development, HarmPBP is first expressed about 4 days prior to adult eclosion and rises to a plateau 2 days prior to adult eclosion. In order to obtain sufficient PBP for further determining its biochemical and physiological properties, a bacterical expression vector of PBP was constructed and successfully expressed in Escherichia coli. The recombinant PBP was shown to cross-react with an anti-PBP antiserum from Antheraea polyphemus. Polyclonal antibodies against HarmPBP were used to mark the distribution of the protein in olfactory sensilla. Very strong labeling was observed in the sensillum lymph of the hair lumen and of the sensillum-lymph cavity. In the male, HarmPBP is expressed in sensilla trichodea and not in sensilla basiconica, while in the female, it is expressed both in sensilla basiconica and sensilla trichodea. PMID- 15352153 TI - Catalytic activity and expression of two flavin-containing monooxygenases from Drosophila melanogaster. AB - Two flavin-containing monooxygenase genes occur in the Drosophila genome (named DmFMO-1 and DmFMO-2). Differences exist between these two FMOs in: (1) genomic DNA architecture and predicted post-translational modifications; (2) recombinant protein solubility, activity, and absorbance spectra; and (3) subcellular distribution and developmental transcription/translation profiles in wildtype flies. Characteristic FAD absorbance spectra and strong catalytic competence in methimazole sulfoxidation were observed for recombinant DmFMO-2. Alternatively, weak sulfoxidation was observed for DmFMO-1, which correlated with reduced solubility in the recombinant system. Western blot analyses using specific antisera raised to each FMO showed the two FMOs to be immunologically distinct. In addition, Western blot analyses revealed FMO protein expression in both the microsomal and cytosolic sub-cellular fractions. Interestingly, a larger form of DmFMO-1 occurs in the cytosol that is most strongly expressed in the adult head. These findings suggest divergent physiological roles for DmFMO-1 and DmFMO-2. More specifically, it appears that DmFMO-1 has a distinct developmental role, while DmFMO-2 may have a general housekeeping function. PMID- 15352154 TI - Characterization of an ecto-phosphatase activity in malpighian tubules of hematophagous bug Rhodnius prolixus. AB - We have characterized a phosphatase activity present on the external surface of intact Malpighian tubules in Rhodnius prolixus. This phosphatase hydrolyses the substrate p-nitrophenyl phosphate at a rate of 3.38 +/- 0.07 nmol Pi x mg(-1) x min(-1). Phosphatase activity decreased with the increase of the pH from 6.4 to 7.6 pH, a range in which tubules cellular integrity was maintained for at least 1 h. Classical inhibitors of acid phosphatase, such as ammonium molybdate, fluoride, vanadate, mpV-PIC, and bpV-PHEN, caused different patters of inhibition. The ecto-phosphatase present an apparent Km of 1.67 +/- 0.34 mM and Vmax of 5.71 +/- 0.37 nmol Pi x mg(-1) x min(-1) for p-NPP. Zinc chloride inhibited 78.2% of ecto-phosphatase activity, with Ki of 0.35 mM. Such inhibition was reversed by incubation with cysteine and GSH, but not DTT, serine, and GSSG, showing that cysteine residues are important for enzymatic activity. Phosphatase activity increased 141% three days after blood meal, and returned to basal levels 2 days later. These results suggest that ecto-phosphatase activity could be involved in a diuretic mechanism, essential in the initial days after a blood meal for the control of Rhodnius homeostasis. PMID- 15352155 TI - Heparin effect on osteoblast-like cells in vitro. PMID- 15352156 TI - Effects of phosphorylation by protein kinase CK2 on the human basal components of the RNA polymerase II transcription machinery. AB - We have investigated the role of phosphorylation by vertebrate protein kinase CK2 on the activity of the General Transcription Factors TFIIA, TFIIE, TFIIF, and RNAPII. The largest subunits of TFIIA, TFIIE, and TFIIF were phosphorylated by CK2 holoenzyme. Also, RNA polymerase II was phosphorylated by CK2 in the 214,000 and 20,500 daltons subunits. Our results show that phosphorylation of TFIIA, TFIIF, and RNAPII increase the formation of complexes on the TATA box of the Ad MLP promoter. Also, phosphorylation of TFIIF increases the formation of transcripts, where as phosphorylation of RNA polymerase II dramatically inhibits transcript formation. Furthermore, we demonstrate that CK2 beta directly interacts with RNA polymerase II, TFIIA, TFIIF, and TBP. These results strongly suggest that CK2 may play a role in regulating transcription of protein coding genes. PMID- 15352157 TI - Ub on the move. AB - Ubiquitination is an increasingly common post-translation modification that controls both the expression and activity of numerous proteins in the eukaryotic cell. One frequent target of the ubiquitin (Ub) modification machinery is transcription factors. Although ubiquitination generally modulates their function by inducing proteasome-dependent degradation, past and recent studies indicate that ubiquitination also regulates nuclear-cytoplasmic trafficking of transcriptional regulators. Ubiquitination is known to modulate transcription factor localization by destroying sequestering proteins and by directly promoting nuclear import and export of modified substrates. This review discusses old and new paradigms relating Ub modification and the control of transcription factor shuttling in and out of the nucleus. PMID- 15352158 TI - Regulation of cell signalling cascades by steroid hormones. AB - Steroid hormones control a wide variety of cellular functions important for cell homeostasis, proliferation, differentiation, and apoptosis. Evidence collected in the last few years indicates that this regulation is mediated by a complex interface of direct control of gene expression (so-called "transcriptional" action, driven by receptors that are localized in cell nucleus) and by regulation of cell signaling/phosphorylation cascades ("extranuclear" action, mediated by receptors that are localized in close proximity to cellular membrane). Regulation of gene expression takes place via ligand-dependent receptor binding to target gene promoters as part of the preinitiation complex, which leads to chromatin remodeling and ultimately regulates the rate of gene expression. Steroid-mediated regulation of cell signaling does not depend on gene transcription or protein biosynthesis. The molecular mechanism of this phenomenon is not fully understood. This review will focus on recent developments related to our understanding of the molecular mechanism of the extranuclear action of steroid hormones. PMID- 15352159 TI - Incorporating pathologists' criteria of malignancy into the evolutionary model for cancer development. AB - A wide variety of alterations in cell and tissue structure still form the basis for cancer diagnosis by pathologists. Cancer development is recognized to be an evolutionary process [Foulds, 1954; Cairns, 1975; Nowell, 1976; Sager, 1982; Tomlinson et al., 1996; Cahill et al., 1999; Tomlinson and Bodmer, 1999], but the phenotypic changes diagnostic of cancer (pathologists' "criteria of malignancy") have not been integrated into the existing evolutionary framework. Since phenotypic changes bear an important relationship to the genetic and physiologic changes underlying Darwinian evolution, we propose that diagnostic structural alterations also bear an important and predictable relation to both the cancer genes and the functional alterations active at any particular step in the development of a cancer. Cancer genes are predicted to mediate the acquisition of cellular-level diagnostic criteria and the diagnostic cellular-level structural changes should reflect in a useful manner the altered cell physiology required for the cell to achieve increased "cellular fitness" at any particular step of colonal evolution. Tissue-level criteria of malignancy should relate less directly to specific cancer genes, but tissue-level criteria should still provide essential insight into the interplay of the altered cellular fitness with the constraints imposed by the cells' microenvironment. The evolutionary framework allows tissue-level criteria of malignancy to be expressed in terms of viable hypotheses for the mechanism of clonal expansion at any particular step in cancer development. This approach to conveying the tissue-level criteria of malignancy complements pattern recognition approaches to diagnosis, and establishes common ground between pathology and cell biology. When viewed from this perspective, the functions of cancer genes appear quite different from those predicted by the "Gatekeeper, Caretaker" or "Hallmarks of Cancer" models. Finally, a full evolutionary framework incorporating the criteria of malignancy restores congruity between the histogenetic classification and the emerging molecular classification of cancer. PMID- 15352160 TI - Human cytomegalovirus pp71: a new viral tool to probe the mechanisms of cell cycle progression and oncogenesis controlled by the retinoblastoma family of tumor suppressors. AB - The DNA tumor virus oncogenes (adenovirus E1A, simian virus 40 (SV40) large T antigen, and papillomavirus E7) have been instrumental in illuminating the molecules and mechanisms of cell cycle progression and carcinogenesis. However, since these multifunctional proteins target so many important cellular regulators, it is sometimes difficult to establish the functional importance of any individual interaction. Perhaps a herpesvirus protein, newly defined as a cell cycle regulator, can help address these issues. Like the DNA tumor virus proteins, the human cytomegalovirus (HCMV) pp71 protein contains a retinoblastoma protein (Rb) binding motif (LxCxD), and stimulates DNA synthesis in quiescent cells. Unlike E1A, T antigen, and E7, pp71 expression does not induce apoptosis, nor does it cooperate to transform primary cells. Determining how pp71 induces cell cycle progression without invoking apoptosis or leading to cellular transformation may help in defining the signals that ultimately lead to these processes. PMID- 15352161 TI - Mechanisms regulating the origins of the vertebrate vascular system. AB - In order to sustain growth, differentiation, and organogenesis, vertebrate embryos must form a functional vascular system early in embryonic development. Intrinsic interest in this process as well as the promise of potential clinical applications has led to significant progress in understanding the mechanisms governing the formation of the vascular system, however the earliest stages of vascular development--the emergence of committed endothelial precursors from the mesoderm--remain unclear. A review of the current literature reveals an unexpected diversity and heterogeneity with respect to where vascular endothelial cells originate in the embryo, when they become committed and the mechanisms governing how endothelial cells acquire their identity. Spatially, a widespread region of the early mesoderm possesses the ability to give rise to vascular endothelial cells; temporally the process is not limited to a small window during embryogenesis, but rather, may continue throughout the lifespan of the organism. On the molecular level, recent findings point to several determinative pathways that regulate, modulate, and extend the scope of the Flk1/VEGF signaling system. An expanding array of novel gene products implicated in endothelial cell type determination appear to act synergistically, with different combinations of factors leading to diverse cellular responses, varying patterns of differentiation, and considerable heterogeneity of endothelial cell types during embryogenesis. PMID- 15352162 TI - Regulation of histone deacetylase activities. AB - Histone deacetylases (HDACs) are enzymes that catalyze the removal of acetyl groups from lysine residues in both histone and non-histone proteins. They play a key role in the regulation of gene transcription and many other biological processes involving chromatin. Significantly, recent studies suggest that HDACs are critically involved in cell-cycle regulation, cell proliferation, differentiation, and in the development of human cancer. HDAC inhibitors currently are being exploited as potential anti-cancer agents. As expected for vital regulators of many cellular processes, the activities of HDACs are tightly controlled and precisely regulated by multiple mechanisms. The activities of most if not all HDACs are regulated by protein-protein interactions. In addition, many HDACs are regulated by post-translational modifications as well as by subcellular localization. Less studied, but perhaps equally important, is the regulation of some HDACs by control of expression, availability of cofactors, and by proteolytic processing. A complete understanding of how HDACs are regulated will contribute not only to our overall knowledge of chromatin structure and gene control, but will offer tremendous insight into approaches for developing therapeutic HDAC inhibitors with improved specificity. PMID- 15352163 TI - Autoinhibited proteins as promising drug targets. AB - Current drug discovery efforts generally focus on a limited number of protein classes, typically including proteins with well-defined catalytic active sites (e.g., kinases) or ligand binding sites (e.g., G protein-coupled receptors). Nevertheless, many clinically important pathways are mediated by proteins with no such obvious targets for small molecule inhibitors. Allosteric inhibitors offer an alternative approach to inhibition of protein activities, particularly for proteins that undergo conformational changes as part of their activity cycle. Proteins regulated by autoinhibitory domains represent one broad class of proteins that meets this criterion. In this article, we discuss the potential of autoinhibited proteins as targets for allosteric inhibitors and describe two examples of small molecules that act by stabilizing native autoinhibited conformations of their targets. We propose that proteins regulated by autoinhibition may be generally amenable to allosteric inhibition by small molecules that stabilize the native, autoinhibited fold. PMID- 15352164 TI - Myocardin/MKL family of SRF coactivators: key regulators of immediate early and muscle specific gene expression. AB - Myocardin, megakaryoblastic leukemia-1 (MKL1), and MKL2 belong to a newly defined family of transcriptional coactivators. All three family members bind to serum response factor (SRF) and strongly activate transcription from promoters with SRF binding sites. SRF is required for the serum induction of immediate early genes such as c-fos and for the expression of many muscle specific genes. Consistent with a role in muscle specific gene expression, myocardin is specifically expressed in cardiac and smooth muscle cells while MKL1 and 2 are broadly expressed. Myocardin has particularly been shown to be required for smooth muscle development while MKL1/2 are required for the RhoA signaling pathway for induction of immediate early genes. SRF can be activated by at least two families of coactivators, p62TCF and myocardin/MKL. These factors bind to the same region of SRF such that their binding is mutually exclusive. This provides one mechanism of regulation of SRF target genes by pathways that differentially activate the coactivators. The RhoA pathway appears to activate MKL1 by altering MKL1's binding to actin and causing MKL1's translocation from the cytoplasm to the nucleus. However, this mechanism of activation of the myocardin/MKL family has not been observed in all cell types such that other regulatory mechanism(s) likely exist. In particular, rapid serum inducible phosphorylation of MKL1 was observed. The regulation of this coactivator family is key to understanding how SRF target genes are activated during muscle cell differentiation or growth factor induced cell proliferation. PMID- 15352165 TI - Extremely low frequency electromagnetic fields as effectors of cellular responses in vitro: possible immune cell activation. AB - There is presently an intense discussion if electromagnetic field (EMF) exposure has consequences for human health. This include exposure to structures and appliances that emit in the extremely low frequency (ELF) range of the electromagnetic spectrum, as well as emission coming from communication devices using the radiofrequency part of the spectrum. Biological effects of such exposures have been noted frequently, although the implication for specific health effects is not that clear. The basic interaction mechanism(s) between such fields and living matter is unknown. Numerous hypotheses have been suggested, although none is convincingly supported by experimental data. Various cellular components, processes, and systems can be affected by EMF exposure. Since it is unlikely that EMF can induce DNA damage directly, most studies have examined EMF effects on the cell membrane level, general and specific gene expression, and signal transduction pathways. In addition, a large number of studies have been performed regarding cell proliferation, cell cycle regulation, cell differentiation, metabolism, and various physiological characteristics of cells. Although 50/60 Hz EMF do not directly lead to genotoxic effects, it is possible that certain cellular processes altered by exposure to EMF indirectly affect the structure of DNA causing strand breaks and other chromosomal aberrations. The aim of this article is to present a hypothesis of a possible initial cellular event affected by exposure to ELF EMF, an event which is compatible with the multitude of effects observed after exposure. Based on an extensive literature review, we suggest that ELF EMF exposure is able to perform such activation by means of increasing levels of free radicals. Such a general activation is compatible with the diverse nature of observed effects. Free radicals are intermediates in natural processes like mitochondrial metabolism and are also a key feature of phagocytosis. Free radical release is inducible by ionizing radiation or phorbol ester treatment, both leading to genomic instability. EMF might be a stimulus to induce an "activated state" of the cell such as phagocytosis, which then enhances the release of free radicals, in turn leading to genotoxic events. We envisage that EMF exposure can cause both acute and chronic effects that are mediated by increased free radical levels: (1) Direct activation of, for example macrophages (or other cells) by short-term exposure to EMF leads to phagocytosis (or other cell specific responses) and consequently, free radical production. This pathway may be utilized to positively influence certain aspects of the immune response, and could be useful for specific therapeutic applications. (2) EMF-induced macrophage (cell) activation includes direct stimulation of free radical production. (3) An increase in the lifetime of free radicals by EMF leads to persistently elevated free radical concentrations. In general, reactions in which radicals are involved become more frequent, increasing the possibility of DNA damage. (4) Long-term EMF exposure leads to a chronically increased level of free radicals, subsequently causing an inhibition of the effects of the pineal gland hormone melatonin. Taken together, these EMF induced reactions could lead to a higher incidence of DNA damage and therefore, to an increased risk of tumour development. While the effects on melatonin and the extension of the lifetime of radicals can explain the link between EMF exposure and the incidence of for example leukaemia, the two additional mechanisms described here specifically for mouse macrophages, can explain the possible correlation between immune cell system stimulation and EMF exposure. PMID- 15352166 TI - Multiple functions of BMPs in chondrogenesis. AB - The ability of bone morphogenetic proteins (BMPs) to promote chondrogenesis has been investigated extensively over the past two decades. Although BMPs promote almost every aspect of chondrogenesis, from commitment to terminal differentiation is well known, the mechanisms of BMP action in discrete aspects of endochondral bone formation have only recently begun to be investigated. In this review, we focus on in vivo studies that have identified interactions between BMP signaling pathways and key downstream targets of BMP action in chondrogenesis. We also discuss evidence regarding the potential roles of BMP receptors in mediating distinct aspects of chondrogenesis, and studies investigating the intersection of BMP pathways with other pathways known to coordinate the progression of chondrocytes through the growth plate. These studies indicate that both Smad-dependent and -independent BMP pathways are required for chondrogenesis, and that BMPs exert essential roles via regulation of the Indian hedgehog (IHH)/parathyroid hormone-related protein (PTHrP) and fibroblast growth factor (FGF) pathways in the growth plate. PMID- 15352167 TI - Thiol oxidation of cell signaling proteins: Controlling an apoptotic equilibrium. AB - Studies of cell signal transduction have predominantly focused on regulation of protein function by phosphorylation. However, recent efforts have begun to uncover another layer of regulation mediated by direct oxidation of cysteine residues in signaling proteins. Typically induced during signaling responses accompanied by generation of reactive oxygen species, these thiol modifications have a variety of functional consequences for target proteins. Using specific signaling protein targets as examples, we discuss how thiol oxidation generally activates pro-apoptotic signaling pathways while inhibiting pathways that promote cell survival. We propose a model in which thiol oxidation acts to control the equilibrium between survival and apoptosis, fine tuning cellular responses that play a central role in the apoptotic decision-making process. We identify areas of focus for future work, including a better understanding of specificity in thiol oxidation events, and a critical need for approaches to examine these modifications under physiologically relevant signaling conditions. PMID- 15352168 TI - Analysis of mesenchymal cells derived from an chondrodysplasia punctuate patient and donors. AB - Conradi-Hunermann syndrome (CDPX2) is X-linked dominant disorder appeared with aberrant punctuate calcification. The skeletal cells derived from the marrow stroma are active in maintaining the skeletal formation. We obtained mesenchymal stem cells from a patient with CDPX2 and studied the formation of colony forming unit-fibroblast (CFU-F) in vitro in comparison cells obtained from normal donors. Cultured cells were studied morphologically and subjected to gene expression analysis. Marrow stromal cells (MSC)-chondrodysplasia punctuate (CDP) cells from CDPX2 were identified by their mosaic morphology formed three phenotypically distinct types of CFU-F colonies. One type consisted of normal fibroblasts with developed cell body and cellular processes; the second type contained pathological small cells without processes; and the third type comprised of mixed cells. We compared gene expression by the MSC-CDP to cells from normal donors. Transcription factors analyzed proliferation potential were similar in both normal and mixed colonies of MSC-CDP and similar to normal MSCs. The message expression for cytokines and extra cellular matrix (ECM) proteins revealed similar expression for biglycan, osteocalcin, and osteonectin, while IL-6, IL-11, and M-CSF mRNA levels were significantly higher in normal cells than in MSC-CDP. Mixed cells had elevated levels for IL-6 and M-CSF mRNA, but expressed IL-11 at the normal range. The studied genes were expressed at lower levels by the pathological (MSC-CDP) cells compared to normal ones. Hence, MSC-CDP was demonstrated to display abnormal morphology and transcription of several investigated genes. This study further illuminates the basis of the mosaic pattern of mesenchymal cells derived from a patient affected with CDPX2, and their gene expression involvement. PMID- 15352169 TI - Post-transcriptional control of expression of sFlt-1, an endogenous inhibitor of vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) is a major modulator of angiogenesis. Biological effects of VEGF are mediated by endothelial cell-surface receptors, KDR and Flt-1. Alternative Flt-1 RNA processing, involving retention of intron 13 and the use of intronic cleavage-polyadenylation signals, produces a secreted form of Flt-1, "sFlt-1," that binds VEGF with high affinity and can inhibit VEGF signaling. To probe mechanisms controlling sFlt-1 expression, we have cloned and sequenced Flt-1 intron 13 from a mouse genomic library and located RNA processing signals potentially involved in sFlt-1 mRNA formation. A minigene construct containing Flt-1 intron 13 directed the expression of both secreted (i.e., cleaved/polyadenylated) and transmembrane (i.e., spliced) forms of Flt-1 mRNA and protein. Using rapid amplification of 3' cDNA ends (3'-RACE) and quantitative PCR (QPCR) analysis to test the activity of intronic cleavage-polyadenylation signals, we observed that multiple sites were utilized for sFlt-1 mRNA processing in both native tissues and Flt-1 minigene transfectants. In transfectants, the most distal signal was utilized preferentially. The potential for interaction between pathways leading to sFlt-1 or full-length Flt-1 was evaluated using QPCR to measure relevant mRNAs after transfection with signal mutants. Decreased expression of sFlt-1 mRNA in cleavage-polyadenylation mutants was accompanied by reciprocal increases in full-length Flt-1 mRNA. Multiple sFlt-1 mRNA species are formed that differ by up to 3.9 kb in their 3'-untranslated regions (UTRs), which contain sites of potential regulatory importance. The reciprocity between sFlt-1 and Flt-1 mRNA expression suggests a novel post-transcriptional mechanism by which sFlt-1 protein production and, thereby, responsiveness to VEGF, may be modulated. PMID- 15352170 TI - Increased apoptosis of periprostatic adipose tissue in VDR null mice. AB - The vitamin D receptor (VDR) is a member of the steroid/retinoid receptor superfamily of nuclear receptors that controls mineral ion homeostatis and has potential tumor-suppressive functions for various cancer types, specifically prostate cancer. A VDR ablated transgenic animal model (VDDRII, vitamin D dependent rickets type II) has been developed and the animals typically have various diseases including, hypocalcemia, hyperparathyroidism, rickets, osteomalacia, and alopecia. This transgenic mouse system provides us with a model to decipher the influences of the VDR on prostatic growth and function. VDRs are abundant both in prostatic epithelial and stromal cells, and vitamin D signaling can be studied in this model. Although, there were no gross differences between the prostate tissue of the experimental and control groups, VDR null mice showed fat necrosis and individual cell apoptosis in the periprostatic adipose tissue. This indicates a possible role of VDR in the signaling pathways resulting the prostate. This may be particularly attractive for VDR targets for the inhibition of cancer progression using VD(3) and its analogs as potential chemo-preventive agents. PMID- 15352171 TI - Effects of hydrogen peroxide on MAPK activation, IL-8 production and cell viability in primary cultures of human bronchial epithelial cells. AB - The airway epithelium is continuously exposed to inhaled oxidants, including airborne pollutants and cigarette smoke, which can exert harmful proinflammatory and cytotoxic effects. Therefore, the aim of our study was to investigate, in primary cultures of human bronchial epithelial cells (HBEC), the signal transduction pathways activated by increasing concentrations (0.25, 0.5, and 1 mM) of hydrogen peroxide (H(2)O(2)), as well as their effects on IL-8 production and cell viability. The reported results show that H(2)O(2) elicited, in a concentration-dependent fashion, a remarkable increase in phosphorylation dependent activation of mitogen-activated protein kinases (MAPKs), associated with a significant induction of IL-8 synthesis and a dramatically enhanced cell death. Pre-treatment of HBEC with MAPK inhibitors was able to significantly inhibit the effects of H(2)O(2) on IL-8 secretion, and to effectively prevent cell death. Therefore, these findings suggest that MAPKs play a key role as molecular transducers of the airway epithelial injury triggered by oxidative stress, as well as potential pharmacologic targets for indirect antioxidant intervention. PMID- 15352172 TI - Hypoxia-induced erythropoietin expression in human neuroblastoma requires a methylation free HIF-1 binding site. AB - The glycoprotein hormone Erythropoietin (EPO) stimulates red cell production and maturation. EPO is produced by the kidneys and the fetal liver in response to hypoxia (HOX). Recently, EPO expression has also been observed in the central nervous system where it may be neuroprotective. It remained unclear, however, whether EPO is expressed in the peripheral nervous system and, if so, whether a neuronal phenotype is required for its regulation. Herein, we report that EPO expression was induced by HOX and a HOX mimetic in two cell lines derived from neuroblastoma (NB), a tumor of the peripheral nervous system. Both cell lines with inducible EPO expression, SH-SY5Y and Kelly cells, expressed typical neuronal markers like neuropeptide Y (NPY), growth-associated protein-43 (GAP 43), and neuron-specific enolase (ENO). NB cells with a more epithelial phenotype like SH-SHEP and LAN-5 did not show HOX inducible EPO gene regulation. Still, oxygen sensing and up-regulation of hypoxia-inducible factor-1 (HIF-1) were intact in all cell lines. We found that CpG methylation of the HIF binding site (HBS) in the EPO gene 3' enhancer was only present in the SH-SHEP and LAN-5 cells but not in SH-SY5Y and Kelly cells with regulated EPO expression. The addition of recombinant EPO to all NB cells, both under normoxic and hypoxic conditions, had no effect on cell proliferation. We conclude that the ability to respond to HOX with an increase in EPO expression in human NB may depend on CpG methylation and the differentiation status of these embryonic tumor cells but does not affect the proliferative characteristics of the cells. PMID- 15352174 TI - Inhibition of cell proliferation and the action mechanisms of arsenic trioxide (As2O3) on human breast cancer cells. AB - Arsenic trioxide (As(2)O(3)) is one of the arsenic compounds found in nature. As(2)O(3) has recently been used to treat patients suffering from retinoic acid receptor (AML). It is of clinical interest to investigate whether As(2)O(3) is also effective in treating solid tumors. Here, we report that As(2)O(3) exhibited inhibitory effects on the proliferation of human breast cancer MCF-7 cells in a dose- and time-dependent manner. The 50% inhibitory concentration (IC(50)) of As(2)O(3) in inhibiting proliferation of MCF-7 cells were 8, 1.8, and 1.2 microM upon 1-, 2-, and 3-day treatment, respectively. In elucidating the underlying action mechanisms, the results of experiments concerning DNA fragmentation and externalization indicated that As(2)O(3) exerted its action on MCF-7 cells via apoptosis, whereas the result of flow cytometry also indicated that As(2)O(3) could induce mitochondrial mediated cell-cycle arrest at G(1) phase. Further studies by Western blot analysis indicated that As(2)O(3) regulated apoptosis and the expression of cell-cycle-related proteins as it upregulated p53 protein level and downregulated bcl-2 protein level. Results in present study indicated that As(2)O(3) might also be a good candidate for treating breast cancer. PMID- 15352173 TI - Paeoniflorin induces apoptosis of lymphocytes through a redox-linked mechanism. AB - Paeoniflorin (PF), isolated from paeony root, has been used as a herbal medicine for more than 1,200 years in China, Korea, and Japan for its anti-allergic, anti inflammatory, and immunoregulatory effects. In this study, we found that PF induces apoptosis in both murine T-lineage cells and human T-cell leukemia Jurkat cells. This apoptosis was mediated through the reduction of mitochondrial membrane potential, activation of caspase, and fragmentation of DNA. Interestingly, PF induced generation of reactive oxygen species (ROS) and a reducing agent, dithiothreitol (DTT), and a ROS scavenger, N-acetyl cysteine (NAC), successfully attenuated the PF-induced apoptosis. Additionally, PF induced the phosphorylation of three mitogen-activated protein (MAP) family kinases, extracellular signal-regulated kinase, c-Jun amino-terminal kinase (JNK), and p38 MAP kinase. Curcumin, an anti-oxidant and JNK inhibitor, inhibited PF-induced apoptosis, suggesting the possible involvement of curcumin-sensitive JNK or other redox-sensitive elements in PF-induced apoptosis. These results partially explain the action mechanism of PF-containing paeony root as a herbal medicine. PMID- 15352175 TI - Non-thermal effects of electromagnetic fields at mobile phone frequency on the refolding of an intracellular protein: myoglobin. AB - Non-thermal effects induced by exposure to microwave electromagnetic field (MW EMF) at 1.95 MHz, a frequency used in mobile communication, have been observed on the refolding kinetics of the heme binding site in an intracellular protein: tuna myoglobin, starting from acidic conditions. We have selected myoglobin because it can be considered a good model to study protein interactions with MW-EMF for its well-known high-resolution crystallographic structure. Myoglobin solutions at pH 3.0 were subjected to 3 h exposure to microwave field (with a specific absorption rate of 51 +/- 1 mW/g); the heme site refolding has been followed by measuring the molecular absorption in the Soret spectral region and the data were fitted to a bi-exponential model. The kinetics of exposed samples appear to be slowered by MW-EMF action. Moreover, the tryptophanyl lifetime distribution of the exposed protein, as deduced by the analysis of the fluorescence emission decay from its single tryptophan, appears sharper if compared to non-exposed protein samples. This observation suggests that the presence of MW-EMF could affect the propensity of protein molecules to populate specific conformational substates among which myoglobin molecules fluctuate at acidic pH. Changes in the structural fluctuation caused by MW perturbation can affect differently the aggregation process that occurs competitively during the protein folding, so representing a potential risk for protein "misfolding." These data suggest that MW-EMF could have also biochemical and, consequently, biological effects on eukaryotic cells that are still under investigation. PMID- 15352176 TI - Prolactin increases HO-1 expression and induces VEGF production in human macrophages. AB - The pituitary hormone prolactin (PRL) is a multifunctional polypeptide which exerts a role on cell proliferation and may also contribute to cell differentiation. PRL is also produced by immune cells and is regarded as a key component of the neuroendocrine-immune loop and as a local regulator of macrophage response. The involvement of PRL in regulating monocyte/macrophage functions is suggested by the presence of PRL receptors in these cells. It has been shown that PRL possess both angiogenic and antiangiogenic effects. Recently, we revealed that augmentation of HO-1 activity enhances PRL-mediated angiogenesis in human endothelial cells. Since macrophages are key participants in angiogenesis our objective was to investigate the effect of PRL also in human macrophages. In vitro treatment of macrophages with PRL was found to increase both heme oxygenase-1 (HO-1) expression and protein synthesis in a time and dose dependent manner as quantified respectively by reverse-transcriptase real-time polymerase chain reaction and Western blot analysis. PRL-treated macrophages also showed an enhanced release of vascular endothelial growth factor (VEGF) as demonstrated by ELISA assay. Furthermore, to determine whether PRL-induced HO-1 activity was required for VEGF production by macrophages, the effect of PRL on the induction of VEGF was studied in the presence of an inducer stannic chloride (SnCl(2)) and of an inhibitor stannic mesoporphyrin (SnMP) of HO activity. Our observations suggest that PRL may regulate monocyte activation and influences not only immune function but also angiogenesis. PMID- 15352177 TI - Homocysteine induces metalloproteinase and shedding of beta-1 integrin in microvessel endothelial cells. AB - Although studies have suggested microvessel endothelial cells (MVEC) activation and induction of matrix metalloproteinases (MMPs) by homocysteine (Hcy), the transduction mechanism leading to endothelial activation was unclear. We hypothesized that Hcy induced metalloproteinase and altered the levels of integrin in MVEC. MVEC from mouse brain were isolated and characterized by CD-31 (PECAM-1) FITC labeling. The MVEC were activated with different doses (6-40 microM) of Hcy. The cultured-conditioned-medium was analyzed for MMP activity by gelatin gel-zymography. TIMP-1, -4, beta-1 integrin, and a disintegrin and metalloproteinase-12 (ADAM-12) were quantified by Western blot analysis. We used MVEC in cell culture to study the effect of increasing concentrations of Hcy upon the secretion of various proteins into the culture medium. MMP-9, beta-1 integrin, ADAM-12, and TIMP-1 were found in increased concentrations in the culture medium of Hcy-treated cells whereas TIMP-4 was decreased. We have shown that purified TIMP-4 blocked the increase of beta-1 integrin shedding in Hcy treated cells. Interestingly, our results suggest that TIMP-1 and TIMP-4 function antagonistically in Hcy-induced signaling pathways. PMID- 15352181 TI - On the TEM and AFM evidence of zeosil nanoslabs present during the synthesis of silicalite-1. PMID- 15352180 TI - A radical show. PMID- 15352183 TI - Towards perfect catalytic asymmetric synthesis: dual activation of the electrophile and the nucleophile. AB - The design and development of new high-performance catalysts for applications in asymmetric catalytic reactions is of ongoing interest in organic chemistry. The combination of a Lewis acid and a Lewis base working in concert is now considered state of the art in stereoselective syntheses. The synergistic activation by two or more reactive centers allows high reaction rates and excellent transfer of stereochemical information. Despite the self-quenching reaction between Lewis acids and Lewis bases that might lead to an inactive catalyst, considerable effort has been directed towards the development of the dual-activation concept. The ultimate goal is to mimic nature by the discovery of catalytic systems analogous to enzymatic processes that involve metal-ion cocatalysts. With this aim, the dual activation concept greatly broadens the range of artificial catalysts. The most efficient catalytic systems are reviewed, and the mechanisms of action are discussed. PMID- 15352184 TI - Regio- and enantioselective iridium-catalyzed allylic alkylation with in situ activated P,C-chelate complexes. PMID- 15352185 TI - Total synthesis of apoptolidin. PMID- 15352186 TI - Fully metalated silsesquioxanes: building blocks for the construction of catalyst models. PMID- 15352187 TI - Sn-filled single-crystalline wurtzite-type ZnS nanotubes. PMID- 15352188 TI - Formation of a stable, lattice-framework disilene: a strategy for the construction of bulky substituents. PMID- 15352189 TI - A combinatorial approach to catalytic peptide dendrimers. PMID- 15352190 TI - Liquid crystals with complex superstructures. PMID- 15352191 TI - Chemical synthesis of the second messenger nicotinic acid adenine dinucleotide phosphate by total synthesis of nicotinamide adenine dinucleotide phosphate. PMID- 15352192 TI - Enantioselective organocatalytic cyclopropanation via ammonium ylides. PMID- 15352193 TI - Simple synthesis of nodulation-factor analogues exhibiting high affinity towards a specific binding protein. PMID- 15352194 TI - Total synthesis of (+/-)-nominine, a heptacyclic hetisine-type aconite alkaloid. PMID- 15352195 TI - Halide recognition through diagnostic "anion-pi" interactions: molecular complexes of Cl-, Br-, and I- with olefinic and aromatic pi receptors. PMID- 15352196 TI - An in vitro assay for evaluation of small-molecule inhibitors of cholesterol absorption. PMID- 15352197 TI - Green and red three-photon upconversion from polymeric lanthanide(III) complexes. PMID- 15352198 TI - Responsive aligned carbon nanotubes. PMID- 15352200 TI - Morphology, structure of dimorphic sperm, and reproduction in the hermaphroditic commensal bivalve Pseudopythina tsurumaru (Galeommatoidea: Kellidae). AB - In Japan Pseudopythina tsurumaru is an up to 10.8 mm-long commensal of the burrowing sea cucumber Protankyra bidentata, whereas in Hong Kong the same species is smaller and associated with the crab Hexapus anfractus, itself a commensal of P. bidentata. Japanese P. tsurumaru is a hermaphrodite tending towards protogyny maturing to a female when > or = 7 mm, and entering the hermaphroditic condition when > or = 9 mm long. In addition to normal euspermatozoa, the species produces 30-32 microm long and 7 x 8 microm broad spindle-shaped paraspermatozoa provided with a conical acrosome, a nucleus, and a bundle of approximately 15-16 flagella issuing from the head region. Paired pouch formed seminal receptacles normally occur in bivalves > or = 6 mm. Bulk sperm transfer presumably takes place by way of spermatozeugmata formed by the two types of sperm cells. Exogenous euspermatozoa attach to particular nonepithelial cells that occupy the interior of the receptacles. These cells, together with their associated sperm, are probably released as syncytial sperm-carrying bodies into the suprabranchial chamber, where the ova are fertilized. PMID- 15352201 TI - Biomechanical modeling and sensitivity analysis of bipedal running ability. I. Extant taxa. AB - I used a simple mathematical model of the inverse dynamics of locomotion to estimate the minimum muscle masses required to maintain quasi-static equilibrium about the four main limb joints at mid-stance of fast running. Models of 10 extant taxa (a human, a kangaroo, two lizards, an alligator, and five birds) were analyzed in various bipedal poses to examine how anatomy, size, limb orientation, and other model parameters influence running ability. I examined how the muscle masses required for fast running compare to the muscle masses that are actually able to exert moments about the hip, knee, ankle, and toe joints, to see how support ability varies across the limb. I discuss the assumptions and limitations of the models, using sensitivity analysis to see how widely the results differed with feasible parameter input values. Even with a wide range of input values, the models validated the analysis procedure. Animals that are known to run bipedally were calculated as able to preserve quasi-static equilibrium about their hindlimb joints at mid-stance, whereas non-bipedal runners (iguanas and alligators) were recognized as having too little muscle mass to run quickly in bipedal poses. Thus, this modeling approach should be reliable for reconstructing running ability in extinct bipeds such as nonavian dinosaurs. The models also elucidated how key features are important for bipedal running capacity, such as limb orientation, muscle moment arms, muscle fascicle lengths, and body size. None of the animals modeled had extensor muscle masses acting about any one joint that were 7% or more of their body mass, which provides a reasonable limit for how much muscle mass is normally apportioned within a limb to act about a particular joint. The models consistently showed that a key biomechanical limit on running ability is the capacity of ankle extensors to generate sufficiently large joint moments. Additionally, the analysis reveals how large ratite birds remain excellent runners despite their larger size; they have apomorphically large extensor muscles with relatively high effective mechanical advantage. Finally, I reconstructed the evolution of running ability in the clade Reptilia, showing that the ancestors of extant birds likely were quite capable runners, even though they had already reduced key hip extensors such as M. caudofemoralis longus. PMID- 15352202 TI - Biomechanical modeling and sensitivity analysis of bipedal running ability. II. Extinct taxa. AB - Using an inverse dynamics biomechanical analysis that was previously validated for extant bipeds, I calculated the minimum amount of actively contracting hindlimb extensor muscle that would have been needed for rapid bipedal running in several extinct dinosaur taxa. I analyzed models of nine theropod dinosaurs (including birds) covering over five orders of magnitude in size. My results uphold previous findings that large theropods such as Tyrannosaurus could not run very quickly, whereas smaller theropods (including some extinct birds) were adept runners. Furthermore, my results strengthen the contention that many nonavian theropods, especially larger individuals, used fairly upright limb orientations, which would have reduced required muscular force, and hence muscle mass. Additional sensitivity analysis of muscle fascicle lengths, moment arms, and limb orientation supports these conclusions and points out directions for future research on the musculoskeletal limits on running ability. Although ankle extensor muscle support is shown to have been important for all taxa, the ability of hip extensor muscles to support the body appears to be a crucial limit for running capacity in larger taxa. I discuss what speeds were possible for different theropod dinosaurs, and how running ability evolved in an inverse relationship to body size in archosaurs. PMID- 15352203 TI - Anatomy and functional morphology of the feeding apparatus of the lesser electric ray, Narcine brasiliensis (Elasmobranchii: Batoidea). AB - Protrusion of the jaws during feeding is common in Batoidea (rays, skates, sawfishes, and guitarfishes), members of which possess a highly modified jaw suspension. The lesser electric ray, Narcine brasiliensis, preys primarily on polychaete annelids using a peculiar and highly derived mechanism for jaw protraction. The ray captures its prey by protruding its jaws beneath the substrate and generating subambient buccal pressure to suck worms into its mouth. Initiation of this protrusion is similar to that proposed for other batoids, in that the swing of the distal ends of the hyomandibulae is transmitted to Meckel's cartilage. A "scissor-jack" model of jaw protrusion is proposed for Narcine, in which the coupling of the upper and lower jaws, and extremely flexible symphyses, allow medial compression of the entire jaw complex. This results in a shortening of the distance between the right and left sides of the jaw arch and ventral extension of the jaws. Motion of the skeletal elements involved in this extreme jaw protrusion is convergent with that described for the wobbegong shark, Orectolobus maculatus. Narcine also exhibits asymmetrical protrusion of the jaws from the midline during processing, accomplished by unequal depression of the hyomandibulae. Lower jaw versatility is a functional motif in the batoid feeding mechanism. The pronounced jaw kinesis of N. brasiliensis is partly a function of common batoid characteristics: euhyostylic jaw suspension (decoupling the jaws from the hyoid arch) and complex and subdivided cranial musculature, affording fine motor control. However, this mechanism would not be possible without the loss of the basihyal in narcinid electric rays. The highly protrusible jaw of N. brasiliensis is a versatile and maneuverable feeding apparatus well-suited for the animal's benthic feeding lifestyle. PMID- 15352204 TI - Spermiogenesis in caecilians Ichthyophis tricolor and Uraeotyphlus cf. narayani (Amphibia: Gymnophiona): analysis by light and transmission electron microscopy. AB - Spermiogenesis, known as spermateleosis in lower vertebrates, is the transformation of the round spermatid into a highly specialized spermatozoon with a species-specific structure. Spermateleosis and sperm morphology of two species of caecilians, Ichthyophis tricolor and Uraeotyphlus cf. narayani, from the Western Ghats of Kerala, India, were studied using light and transmission electron microscopy. Spermateleosis is described in early, mid-, and late phases. During the early phase, the spermatid nucleus does not elongate, but the acrosome vesicle is Golgi-derived and its material is produced as a homogeneous substance rather than as discrete granules. In development of the acrosome, the centrioles shift in position to the lower half of the cell. The acrosomal vesicles take the full shape of the acrosome with the establishment of the perforatorium in midphase. An endonuclear canal develops and accommodates the perforatorium. The incipient flagellum is laid down when the proximal centriole attaches to the posterior side of the nucleus and the distal centriole connects to the proximal centriole, which forms the basal granule of the acrosome. The axial fiber also appears during midphase. The mitochondria shift in position to the posterior pole of the cell to commence establishment of the midphase. Late phase is characterized by nuclear condensation and elongation. Consequently, the final organization of the sperm is established with the head containing the nucleus and the acrosome. The undulating membrane separates the axoneme and axial fiber. Most of the cytoplasm is lost as residual bodies. PMID- 15352205 TI - Functional morphology of the reproductive system of Galathea intermedia (Decapoda: Anomura). AB - Spermatophore formation in Galathea intermedia begins in the proximal part of the vas deferens. The contents subsequently form a spermatophoric ribbon, the so called "secondary spermatophore," in its distal part. A strongly muscular ductus ejaculatorius is present in the coxa of the fifth pereiopod which builds up pressure for the extrusion of the spermatophoric ribbon. After extrusion, the ribbon is caught by the first gonopod, while the second gonopod dissolves the matrix of the ribbon. During copulation the spermatophores are randomly placed on the sternum of the female, near the genital opening, by the fifth pereiopods of the male. Subsequent ovulation of the female via the genital opening, an active process accomplished through muscular activity, results in fertilization of the eggs by the exploding spermatophores. External intersexes are characterized by both male and female external sexual characters, but in all individuals only male gonads are present. No trace of a female reproductive system could be detected. Thus, these external intersexes are exclusively functional males. PMID- 15352206 TI - Comparative anatomy of the superfamily Myliobatoidea (Chondrichthyes) with some comments on phylogeny. AB - The anatomy of species belonging to the superfamily Myliobatoidea was examined with the aim of better determining their phylogenetic relationships. A wide variation among genera was observed in skeletal anatomy, despite the fact that they all share a common morphological pattern. However, variation among species of the same genus was low, excepting Mobula. Dorsal musculature showed a substantial consistency, except for the epiaxialis muscle, which was larger in rhinopterids and mobulids. Variation in the ventral muscles was low among species of the same genus, but considerable among different genera. Mobulids have a reduction in ventral muscles, while rhinopterids and myliobatoids show an increase in muscular mass. A consensus tree shows a basal split into two groups. The first includes the family Gymnuridae with the genera Gymnura and Aetoplatea; this group is supported by seven synapomorphies, including: 27(1) ceratobranchialis fused proximally, 36(1) anterior lateral processes present in the synarcual, 52(0) quadratomandibularis internal muscle present. The second group is composed of the family Myliobatidae (Myliobatis, Aetomylaeus, Aetobatus, Rhinoptera, Mobula, and Manta), this group is supported by 11 synapomorphies, including: 5(1) first postorbital process fused with the second, 21(1) fused mandibular symphysis, 24(1) first hypobranchial cartilage absent, 48(2) epiaxialis muscle inserted in the cranial orbital region, 73(1) pectoral fins joined behind the orbital region. This study concluded that myliobatoids (Myliobatis, Aetomylaeus, and Aetobatus) integrate a monophyletic group which, unlike other phylogenies previously obtained, is the sister group of rhinopterids (Rhinoptera). Mobulids (Mobula and Manta) are the sister group of myliobatoids rhinopterids. PMID- 15352207 TI - Changes in oviductal morphology of the skink, Lampropholis guichenoti, associated with egg production. AB - We describe changes in the morphology of the oviductal epithelium of an oviparous skink, Lampropholis guichenoti, during the course of egg production and oviposition: to characterize the luminal epithelial changes; to provide a baseline for understanding uterine changes in viviparous species; and to establish whether the plasma membrane transformation of uterine epithelial cells is indeed a feature restricted to viviparous species. Oviducts from vitellogenic, gravid, and postgravid females were observed using scanning electron microscopy. Cellular characteristics of the oviductal epithelium previously used to determine the plasma membrane transformation were assessed morphologically. Three anatomically different areas were defined within the oviduct, but no plasma membrane transformation was observed in the oviparous skink, suggesting that this is a phenomenon particular to viviparity. PMID- 15352208 TI - Oxidative stress-induced death in the nervous system: cell cycle dependent or independent? AB - Neuronal death, attributable to perturbed redox homeostasis, is the underlying factor in many acute and chronic neurological disorders. The mechanisms employed by oxidatively stressed neurons to commit to cell death pathways are beginning to be characterized, but this is hampered by a lack of good models that extrapolate readily to redox-dependent neuronal death paradigms. In this Mini-Review, we discuss mechanisms by which oxidative stress can result in neurodegeneration. We examine evidence on which terminally differentiated neurons might commit to death under conditions of oxidative stress. In some cases, death may be linked to an aberrant and uncoordinated reentry into the cell cycle and mitotic catastrophe. Other evidence suggests that cell cycle reentry is not mandatory for death execution. Rather, the reexpression of cell cycle proteins may induce apoptotic pathways in a cell cycle-independent manner. In contrast to these models, there is also evidence that oxidative neuronal death is independent of cell cycle proteins. We conclude that oxidative stress-induced neuronal death may be promoted via several pathways, which may be cycle protein dependent or independent. The determining factor for which or how many pathways are induced appears to be context dependent and determined by the level and duration of oxidative stress. PMID- 15352209 TI - Constitutive Ras activity induces hippocampal hypertrophy and remodeling of pyramidal neurons in synRas mice. AB - The small G protein Ras, which is involved critically in neurotrophic signal transduction, has been implicated in neuronal plasticity of both the developing and the adult nervous systems. In the present study, the cumulative effects of constitutive Ras activity from early in postnatal development into the adult upon the morphology of hippocampal pyramidal neurons were investigated in synRas mice overexpressing Val12-Ha-Ras postmitotically under the control of the rat synapsin I promoter. In synRas mice, stereologic investigations revealed hypertrophy of the hippocampus associated with an increase in perikaryal size of pyramidal neurons within the CA2/CA3 region and the gyrus dentatus. Morphometric analyses of Lucifer Yellow-filled CA1 pyramidal neurons, in addition, demonstrated considerable expansion of dendritic arbors. The increase in basal dendritic size was caused primarily by alterations of intermediate and distal segments and was associated with an enlarged dendritic surface. Apical dendrites showed similar but more moderate changes, which were attributed mainly to elongation of terminal segments. Sholl analyses illustrated higher complexity of both basal and apical trees. Despite significant morphologic alterations, dendritic arbors preserve their major design principles. The synaptic density within the stratum radiatum of CA1 remained unchanged; however, increases in the total hippocampal volume and in apical dendritic size imply an increment in the absolute number of synaptic contacts. The data presented here suggest a critical involvement of Ras dependent signaling in morphoregulatory processes during the maturation and in the maintenance of hippocampal pyramidal neurons. PMID- 15352210 TI - Treatment of cerebellar granule cell neurons with the neurotrophic factor pigment epithelium-derived factor in vitro enhances expression of other neurotrophic factors as well as cytokines and chemokines. AB - Microarray analyses demonstrated that a variety of genes was affected by treatment of cerebellar granule cell neurons with the neurotrophic factor pigment epithelium-derived factor (PEDF). The genes for neurotrophins, glial cell-derived neurotrophic factor (GDNF), and their receptors were regulated differentially in immature versus mature neurons; however, nerve growth factor (NGF), neurotrophin (NT)-3, and GDNF did not contribute to the protective effect of PEDF. Brain derived neurotrophic factor (BDNF) seemed capable of inducing apoptosis, because a blocking antibody enhanced the protective effect of PEDF. In addition, PEDF exposure also stimulated expression of several cytokine and chemokine genes. Removal of the less than 1% of microglia in the cultures by treatment with L leucine methyl ester, combined with enzyme-linked immunosorbent assays (ELISAs), demonstrated that the cerebellar granule cells constitutively produce three chemokines, macrophage inflammatory protein (MIP)-1alpha, MIP-2, and MIP-3alpha, whose production is enhanced further by treatment with PEDF. Blocking antibodies to each of the chemokines was protective under control conditions, suggesting that they may contribute to the "natural" apoptosis occurring in the cultures, and enhanced the effects of PEDF. Although PEDF enhanced production of all three chemokines, the blocking antibodies did not increase its protective effect against induced apoptosis. These results suggest that although PEDF enhances expression of other neurotrophic factors or chemokines, it does not exert its neuroprotective effect on cerebellar granule cells through their production. PMID- 15352211 TI - A sulfatase regulating the migratory potency of oligodendrocyte progenitor cells through tyrosine phosphorylation of beta-catenin. AB - By using cDNA subtraction, we identified an extracellular sulfatase (RsulfFP1) from rat oligodendrocyte progenitor cells (OPCs) whose mRNA expression is down regulated by tumor necrosis factor-alpha. RsulfFP1 mRNA was expressed specifically in the floor plate and the ventral portion of the rat spinal cord at E15. The expression pattern of RsulfFP1 overlapped with the OPCs, which are also located at the ventral region of the ventricular zone. After this stage, RsulfFP1 expression was attenuated, and the OPCs efficiently migrated throughout the spinal cord. The modification of CG-4 cells, a cell line established from rat O2A cells, by RsulfFP1 activated canonical Wnt signaling. Furthermore, the deletion of RsulfFP1 expression by an antisense oligonucleotide caused impairment of OPC migration in rat spinal cord slice culture. Modification of cells by RsulfFP1 resulted in the increased tyrosine phosphorylation of immunoprecipitated beta catenin, suggesting that sulfation of the extracellular matrix induced by this sulfatase might be responsible for an increase in Wnt signaling that is involved in the migration of OPCs. Thus, the present study revealed that a sulfatase is responsible for the migration of OPCs and activates intracellular mechanisms that regulate migration. PMID- 15352212 TI - BDNF overexpression produces a long-term increase in myelin formation in the peripheral nervous system. AB - The neurotrophin brain-derived neurotrophic factor (BDNF) is an endogenous regulator of the myelination process during development in the peripheral nervous system. Enhancement of myelin formation by BDNF is mediated by the neurotrophin receptor p75(NTR). Although this neurotrophin is a positive modulator of myelination during early development, the final effects of BDNF on myelin sheaths after active myelination is completed are largely unknown. Using BDNF transgenic mice, we examined the long-term effects of BDNF on myelination of the peripheral nervous system in vivo. Elevation of BDNF levels in the transgenic mice produced an increase in both the rate and extent of the myelination process. BDNF enhanced and accelerated myelin formation during early development and this increase in myelin content and thickness was maintained in adulthood. Besides enhanced myelination, BDNF also influenced axon caliber size but to a lesser extent. This lagging increase in axon caliber compared to myelin suggests that the axon size is not the only determinant of myelin thickness. PMID- 15352213 TI - T-cell hybridoma specific for myelin oligodendrocyte glycoprotein-35-55 peptide produced from HLA-DRB1*1501-transgenic mice. AB - The goal of this study was to establish an unlimited and standardized source of humanized myelin peptide-specific T cells for in vitro testing of biological function. Thus, we perpetuated myelin oligodendrocyte glycoprotein (MOG)-35-55 peptide-specific T cells obtained from immunized HLA-DRB1*1501-transgenic (Tg) mice by somatic fusions with BW5147 thymoma cells or BW5147 T-cell receptor (TCR) alpha(-)beta(-) variant (BW5147 variant) cells. The resulting T-cell hybridomas responded strongly to both mouse MOG-35-55 (42S) and human MOG-35-55 peptide (42P), regardless of which peptide was used for initial immunization, and were DRB1*1501 restricted. The MOG-35-55-reactive T-cell hybridomas were CD3(+)CD4(+)CD8(-) and expressed intracellular Th1 cytokines upon concanavalin A stimulation. Clones from either human MOG-35-55- or mouse MOG-35-55-selected hybridomas uniquely expressed the TCR BV8 gene in combination with AV17 and AV11 genes. V gene analyses confirmed the expression of TCR AV1, AV11, AV16, BV1, and BV5 gene segments in the widely used fusion partner BW5147 and demonstrated deletion of TCR AV1, AV11, and BV1 in the BW5147 variant. T-cell hybridomas were positively stained with anti-TCR beta-chain antibody on the cell surface, whereas neither BW5147 nor its variant had positive TCR surface expression. For functional application, we found that a monomeric form of the human HLA-DR2 derived recombinant T-cell receptor ligand (RTL) covalently linked to human MOG 35-55 peptide specifically inhibited proliferation of a hybridoma clone selected with human MOG-35-55 but not a different hybridoma clone selected with myelin basic protein. The RTL-induced inhibition in vitro of the human MOG-35-55 peptide specific hybridoma reflected the ability of the RTL to inhibit experimental autoimmune encephalomyelitis induced by human MOG-35-55 peptide in HLA-DR2 transgenic mice. Thus, the MOG-35-55 peptide-specific T-cell hybridoma from DR2 Tg mice represents a novel humanized T-cell reagent useful for standardized biological screening of both DR2-restricted stimulation and RTL-dependent inhibition of response to human MOG-35-55 peptide. PMID- 15352214 TI - Effect of manipulation of iron storage, transport, or availability on myelin composition and brain iron content in three different animal models. AB - Several observations suggest that iron is an essential factor in myelination and oligodendrocyte biology. However, the specific role of iron in these processes remains to be elucidated. This role could be as an essential cofactor in metabolic processes or as a transcriptional or translational regulator. In this study, we used animals models each with a unique defect in iron availability, storage, or transfer to test the hypothesis that disruptions in these mechanisms affect myelinogenesis and myelin composition. Disruption of iron availability either by limiting dietary iron or by altering iron storage capacity resulted in a decrease in myelin proteins and lipids but not the iron content of myelin. Among the integral myelin proteins, proteolipid protein was most consistently affected, suggesting that limiting iron to oligodendrocytes results not only in hypomyelination but also in a decrease in myelin compaction. Mice deficient in transferrin must receive transferrin injections beginning at birth to remain viable, and these mice had increases in all of the myelin components and in the iron content of the myelin. This finding indicates that the loss of endogenous iron mobility in oligodendrocytes could be overcome by application of exogenous transferrin. Overall, the results of this study demonstrate how myelin composition can be affected by loss of iron homeostasis and reveal specific chronic changes in myelin composition that may affect behavior and attempts to rescue myelin deficits. PMID- 15352215 TI - Developmental shift in expression of netrin receptors in the rat spinal cord: predominance of UNC-5 homologues in adulthood. AB - Netrins are a family of secreted proteins required for normal neural development. Netrin-1 is expressed at similar levels in the adult rat spinal cord and the embryonic CNS, suggesting that it contributes to adult CNS function. Here we show that the netrin receptors dcc, neogenin, unc5h1, unc5h2, and unc5h3 are also expressed in the adult rat spinal cord. Lower levels of DCC and neogenin were detected in the adult relative to the embryonic CNS. Conversely, the adult spinal cord contains increased levels of UNC-5 homologues in comparison with the embryo. Multiple mRNA transcripts detected by Northern blot analysis suggested that netrin receptors might be encoded by alternatively spliced mRNAs. We have identified a novel alternatively spliced mRNA encoding UNC5H1, UNC5H1(Delta)TSP1, which lacks the first of the two extracellular thrombospondin domains. This novel splice variant is the major transcript detected in the early embryonic CNS, although both splice variants are expressed in the adult. Previously identified alternatively spliced mRNAs encoding DCC and neogenin were also detected. Dcc, neogenin, unc5h1, unc5h2, and unc5h3 are expressed by subsets of neurons. Robust expression of unc5h2 was found in glia. These findings suggest that unc-5 homologues constitute a major mode of netrin-1 signal transduction in the adult spinal cord and may be involved in phenomena analogous to axon repulsion, such as inhibiting process extension and collateral sprouting. PMID- 15352216 TI - Neutralization of the chemokine CXCL10 enhances tissue sparing and angiogenesis following spinal cord injury. AB - After spinal cord injury, there is a chemoattractant-mediated inflammatory response that is associated with secondary degeneration. The chemoattractant CXCL10 recruits CD4 Th1 cells via the CXCR3A receptor and inhibits growth and chemotaxis of endothelial cells via the CXCR3B receptor. To test the hypothesis that CXCL10 inhibits angiogenesis following spinal cord injury, we assayed the brainstems and spinal cords of spinal cord-injured mice treated with anti-CXCL10 antibodies for expression of angiogenesis-associated genes and quantified blood vessels within their spinal cords. Brainstem microarray analysis indicated eight angiogenesis-associated genes that had significantly higher expression levels in the treated mice than in the untreated mice. Ribonuclease protection assays of the spinal cords showed a significant increase in eight angiogenesis-associated genes in treated animals compared with untreated animals. Histological analysis of the spinal cords of treated and untreated mice showed a significant increase in the number of blood vessels in treated animals. We conclude that CXCL10 plays a critical role in vasculature remodeling following spinal cord injury and that angiogenesis is enhanced following anti-CXCL10 treatment of spinal cord injuries. Improved blood flow and oxygen supply to the injury site may contribute to the functional improvement associated with this treatment. PMID- 15352217 TI - Human and murine lymphocyte neurotrophin expression is confined to B cells. AB - Recent reports indicate that autoreactive T cells may produce neurotrophic factors capable of mediating repair and regeneration of damaged neurons. By using semiquantitative RT-PCR, we examined gene expression of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and the trkB BDNF receptor in autoreactive T cells from SWXJ mice immunized with the p104 117 encephalitogen of myelin proteolipid protein (PLP 104-117). We observed antigen-inducible expression of NGF and BDNF, but not NT-3 and trkB, in lymph node cells activated with PLP 104-117. To determine which leukocyte subpopulation expressed neurotrophins, CD4(+), CD8(+), B220(+), CD11b(+), and NK1.1(+) cells were purified from activated primary cultures, and their mRNAs were analyzed. Neurotrophin expression was also measured in CD3(+) T cells purified from mouse CNS during acute onset of experimental autoimmune encephalomyelitis as well as in resting and activated human T cells and B cells purified from peripheral blood of normal subjects. In all cases, we found that neurotrophin expression was confined exclusively to B cells (B220(+)) in both mouse and human. CD3(+), CD4(+), and CD8(+) T cells as well as NK1.1(+) cells and CD11b(+) monocytes and macrophages did not express any detectable BDNF, NGF, NT-3, or trkB under any conditions. Our data indicate that B cells rather than T cells are the predominant if not the only source of leukocyte-derived neurotrophins and as such may provide "protective autoimmunity" in repair and regeneration of the injured nervous system. PMID- 15352218 TI - Antinociceptive effects of galanin in the rat tuberomammillary nucleus and the plasticity of galanin receptor 1 during hyperalgesia. AB - Although the tuberomammillary nucleus (TM) is well defined in terms of anatomy and neurochemistry, little is known about its function in nociceptive modulation. There was an abundance of galanin-immunoreactive fibers in the TM, and galanin has been implicated in pain processing. The present study assessed the role of galanin in the modulation of nociception in the TM of rats. Intra-TM injection of galanin dose-dependently increased the hindpaw withdrawal latency of rats to a noxious thermal stimulus, indicating an antinociceptive role of galanin in the TM. The antinociceptive effect of galanin was blocked by a subsequent intra-TM injection of galantide, a putative galanin receptor antagonist, suggesting that the antinociceptive effect of galanin is mediated by galanin receptors. Moreover, there was abundant galanin receptor 1 (GalR1) in the TM, and the number of GalR1 positive neurons in the ipsilateral TM increased significantly after unilateral loose ligation of the sciatic nerve compared with the contralateral TM or the TM of intact rats. However, the number of GalR1-positive neurons was not significantly altered by carrageenan-induced inflammation, in either the ipsilateral or the contralateral TM. The results suggest that galanin and GalR1 in the TM may play important roles in pain regulation. PMID- 15352219 TI - Differential roles for neuropeptide Y Y1 and Y5 receptors in anxiety and sedation. AB - Central administration of neuropeptide Y (NPY) causes both anxiolysis and sedation. Previous studies suggest that both effects are mediated via NPY Y1 receptors. However, most of these studies were carried out before the advent of specific NPY receptor ligands. Therefore, a potential role for other NPY receptors in anxiety and sedation remains a possibility. In the present study, we addressed this issue by testing the effects of intracerebroventricular (i.c.v.) injection of NPY as well as specific receptor agonists for the Y1 receptor ([D His(26)]NPY), Y2 receptor (C2-NPY), and Y5 receptor ([cPP(1-7),NPY(19 23),Ala(31),Aib(32),Gln(34)]hPP) in the elevated plus maze and open field tests. As with NPY, the Y1 agonist had a dose-dependent anxiolytic-like effect in both behavioral tests. In contrast to NPY, which caused significant sedation in the open field test, the Y1 agonist was without sedative effect. The Y2 agonist showed neither anxiolytic-like nor sedative effects. The Y5 agonist showed anxiolytic-like activity in both behavioral tests and caused sedation in the same dose range as NPY in the open field test. These results indicate that anxiolytic like effects of i.c.v.-administered NPY in rats are mediated via both Y1 and Y5 receptors, whereas sedation is mediated via Y5 receptors. PMID- 15352220 TI - Role of N-methyl-D-aspartate receptors in gastric mucosal blood flow induced by histamine. AB - Ionotropic N-methyl-D-aspartate (NMDA) receptor agonists, L-aspartic acid (L-Asp) and NMDA, have been shown to inhibit histamine-stimulated acid secretion, but their effect on gastric mucosal blood flow (GMBF) is largely unknown. The aim of this study was to investigate whether L-Asp and NMDA inhibit histamine-stimulated GMBF and to examine the expression patterns of NMDA receptor subunits NR1, NR2A, and NR2B in rat stomach. Laser Doppler flowmetry was used to measure gastric blood flow in anesthetized rats. The GMBF was assessed during an intravenous infusion of histamine in the presence of tripelennamine. The effects of L-Asp and NMDA on histamine-induced gastric blood flow were examined. In addition, the distribution patterns of NR1-, NR2A-, and NR2B-contaning NMDA receptors in rat stomach were determined immunohistochemically by using specific antibodies against NR1, NR2A, and NR2B. Histamine-induced enhancement of GMBF depended on acid secretion and the activation of H(2)-receptors. Neither L-Asp nor NMDA had an effect on the spontaneous GMBF. However, L-Asp and NMDA reduced the histamine induced increase in GMBF. DL-2-amino-5-phosphonopentanoic acid (AP-5), an NMDA receptor antagonist; and prazosin, an alpha(1)-receptor antagonist; but not propanolol, a beta(2)-receptor antagonist; or yohimbine, a alpha(2)-receptor antagonist; reversed the inhibitory effect of L-Asp and NMDA on the histamine induced increase in GMBF. Therefore, L-Asp and NMDA inhibit histamine-induced GMBF via a mechanism involving the activation of NMDA receptors and alpha(1)- adrenoceptors. The fact that NMDA receptor subunits NR1, NR2A, and NR2B were found to be localized in the rat stomach as visualized immunohistochemically with specific antibodies against NR1, NR2A, and NR2B is consistent with this hypothesis. PMID- 15352221 TI - Repeated contact with subtoxic soman leads to synaptic vulnerability in hippocampus. AB - Soman, an anticholinesterase and dangerous nerve agent, produces convulsions, memory impairment, and cell loss in the brain, especially in the hippocampus. Soman-induced accumulation of acetylcholine initiates mechanisms responsible for the development of incapacitating seizures. The prolonged epileptiform nature of these seizures causes the release of another excitatory neurotransmitter, glutamate, which has been linked to the toxic action of the nerve agent. Here, we tested whether subtoxic soman exposures influence the brain's sensitivity to glutamate-based excitotoxicity. Over a 1-week period, hippocampal slice cultures were exposed daily to a transient level of soman that produced no evidence of synaptic deterioration. After the subtoxic soman treatments, however, the tissue became vulnerable to a brief episode of glutamate receptor overstimulation that normally resulted in little or no excitotoxic damage. In those slice cultures treated with subtoxic soman, a decline in synaptic markers as well as an increase in spectrin breakdown occurred 24 hr after the mild excitotoxic event. Exposure to high soman concentrations alone produced similar synaptic degeneration, but without evident cell death, suggesting that synaptic decline is an early neurotoxicological response to the nerve agent. Interestingly, enhanced excitotoxic sensitivity caused the brain tissue to become susceptible to disparate insults initiated before or after the soman contact. These findings indicate that seemingly innocuous soman exposures leave the hippocampus sensitive to the types of insults implicated in traumatic brain injury and stroke. They also warn that asymptomatic contact with soman may lead to progressive synaptopathogenesis and that early indicators of soman exposure are critical to prevent potential brain injury. PMID- 15352222 TI - 6-Hydroxydopamine induces dopaminergic cell degeneration via a caspase-9-mediated apoptotic pathway that is attenuated by caspase-9dn expression. AB - This study showed that primary dopaminergic neurons or the dopaminergic cell line MN9D, when exposed to 15 min of the parkinsonian toxin 6-hydroxydopamine (6-OHDA) in the range of 30-100 microM, underwent delayed degeneration and exhibited hallmarks of apoptosis. These results, along with the absence of any increase in lactate dehydrogenase (LDH) release from the degenerated cells, imply that apoptosis was the dominant mode of cell death. Moreover, a distinct elevation in the measured cellular activities of caspase-9 and -3 but not of caspase-8 points to the caspase-9/caspase-3 cascade as the predominant apoptotic pathway in the degeneration of dopaminergic neurons and MN9D cells. In addition, the presence of caspase-9 or -3 peptide inhibitors but not of caspase-8 inhibitor attenuated cell death significantly, supporting the notion that only the intrinsic apoptotic pathway is utilized to achieve cell death. Finally, overexpression of a mutant caspase-9 with dominant negative phenotype (caspase-9dn) in MN9D cells and primary dopaminergic neurons via the adenovirus and adenoassociated virus gene delivery system, respectively, conferred marked increases in tolerance to the toxicity of 6-OHDA. These results point to the intrinsic caspase-9/caspase-3 cascade as the predominant signaling pathway underlying dopaminergic cell death induced by 6-OHDA and suggest that gene delivery of caspase-9dn can attenuate this pathway and its degenerative consequences. PMID- 15352223 TI - High-resolution magic angle spinning and 1H magnetic resonance spectroscopy reveal significantly altered neuronal metabolite profiles in CLN1 but not in CLN3. AB - The neuronal ceroid lipofuscinoses (NCLs) are among the most severe inherited progressive neurodegenerative disorders of children. The purpose of this study was to compare the in vivo 1.5-T 1H magnetic resonance (MR) and ex vivo 14.3-T high-resolution (HR) magic angle spinning (MAS) 1H MR brain spectra of patients with infantile (CLN1) and juvenile (CLN3) types of NCL, to obtain detailed information about the alterations in the neuronal metabolite profiles in these diseases and to test the suitability of the ex vivo HR MAS (1)H MRS technique in analysis of autopsy brain tissue. Ex vivo spectra from CLN1 autopsy brain tissue (n = 9) significantly differed from those of the control (n = 9) and CLN3 (n = 5) groups, although no differences were found between the CLN3 and the control groups. Principal component analysis of ex vivo data showed that decreased levels of N-acetylaspartate (NAA), gamma-aminobutyric acid (GABA), glutamine, and glutamate as well as increased levels of inositols characterized the CLN1 spectra. Also, the intensity ratio of lipid methylene/methyl protons was decreased in spectra of CLN1 brain tissue compared with CLN3 and control brain tissue. In concordance with the ex vivo data, the in vivo spectra of late-stage patients with CLN1 (n = 3) revealed a dramatic decrease of NAA and a proportional increase of myo-inositol and lipids compared with control subjects. Again, the spectra of patients with CLN3 (n = 13) did not differ from those of controls (n = 15). In conclusion, the ex vivo and in vivo spectroscopic findings were in good agreement within all analyzed groups and revealed significant alterations in metabolite profiles in CLN1 brain tissue but not in CLN3 compared with controls. Furthermore, HR MAS 1H MR spectra facilitated refined detection of neuronal metabolites, including GABA, and composition of lipids in the autopsy brain tissue of NCL patients. PMID- 15352225 TI - A human proteome project with a beginning and an end. AB - Research activities centered on the ensemble of and individual human proteins have taken on numerous guises, some of which fall under the banner of what could be defined as a Human Proteome Project (HPP). However, the latter has yet to take on the apparent global focus of its predecessor, the Human Genome Project. The reasons for this are both financial and technical. The disparate properties afforded to each protein by a 20-letter code render a single unifying approach difficult to implement, while the current limit of analytical detection has yet to deliver an entire proteome for even the simplest of microbes. The situation is complicated further by the fact that low abundance proteins dominate within any living cell. Thus, enhancement of signal-to-noise ratio by affinity ligands becomes of paramount importance if whole-organism proteomics is to be realized. The generation of such ligands (molecules exhibiting desirable affinity and selectivity for target) could provide the necessary focus and a task list with a definable beginning and end. Such a finite task list is considered essential if an HPP might one day deliver global coverage on a scale seen currently for the total DNA sequence of some 200 living organisms. PMID- 15352226 TI - A critical evaluation of sample extraction techniques for enhanced proteomic analysis of recalcitrant plant tissues. AB - Most published proteomics studies of bulk plant tissues use a procedure in which proteins are precipitated with trichloroacetic acid (TCA) and acetone (TCA-A), but few attempts have been made to contrast this approach in a systematic way with alternative methods against a spectrum of tissues. To address this, TCA-A was compared with another acetone-based protocol (TCA-B) or a phenol (Phe)-based method, targeting a range of tomato tissues and three species of fruits that contain high levels of contaminating compounds: banana, avocado and orange. The Phe method gave a higher protein yield and typically greater resolution and spot intensity, particularly with extracts from tissues containing high levels of soluble polysaccharides. The methods also generated remarkably different two dimensional gel electrophoresis (2-DE) protein spot patterns. Peptide mass fingerprinting was used to identify polypeptides that were common to multiple extracts or uniquely present in one extract type. While no clear pattern emerged to explain the basis for the differential protein extraction, it was noted that the Phe method showed enhanced extraction of glycoproteins. These results suggest that the Phe protocol is highly effective with more recalcitrant tissues and that a combination of TCA-A and Phe methods provides enhanced 2-DE based proteomic analyses of most plant tissues. PMID- 15352227 TI - Parallel isoelectric focusing chip. AB - Fast isoelectric focusing (IEF) is becoming a key method in modern protein analysis. We report here the theory and experimental results of new parallel isoelectric devices (PID) for fast IEF. The main separation tool of any PID is a dielectric membrane with conducting channels filled by immobiline gels of varying pH. The pH value of the surrounding aqueous solution is not equal to the pH of any of the channels. The membrane is held perpendicular to the applied electric field. Proteins are collected (trapped) in the channels whose pH values are equal to the pI of the proteins. The fast particle transport between different channels takes place due to convection in the aqueous solution. We developed a mathematical model for PID. Experiment duration is shown to be proportional to the number of different bands N (the peak capacity in standard IEF) in contrast with N(2) for usual IEF devices. This model was validated with experimental results. Parallel IEF accelerates the fractionation of proteins by their pI values (down to several minutes) allowing a more desirable collection efficiency to be achieved. The main theoretical limitation of PID resolution is the sensitivity of proteins to pH change due to the Coulomb blockade effect. The existence of a minimal pH change deltapH(min) for each type of protein is shown: deltapH(min) approximately r(-1) for globular molecules with radius r. PMID- 15352228 TI - A peptide preconcentration approach for nano-high-performance liquid chromatography to diminish memory effects. AB - An advanced method has been developed for the analysis of proteolytic digests of complex protein mixtures by reversed phase high-performance liquid chromatography coupled to mass spectrometry. The occurrence of memory effects was prevented by a parallel set of two precolumns employed for simultaneous separation and washing procedures. The system was tested extensively, and tryptic digests of three single proteins were analyzed. In addition, different solvent systems were evaluated for effective washing of the employed precolumns. Using the analytical strategy presented, a reliable identification of proteins in complex mixtures was obtained and not hampered by the occurrence of memory effects. PMID- 15352229 TI - Automated, on-line two-dimensional nano liquid chromatography tandem mass spectrometry for rapid analysis of complex protein digests. AB - Evaluation of cellular processes and their changes at the level of protein expression and post-translational modifications may allow identification of novel proteins and the mechanisms involved in pathogenic processes. However, the number of proteins and, after tryptic digestion, of peptides from a single cell can be tremendously high. Separation and analysis of such complex peptide mixtures can be performed using multidimensional separation techniques such as two-dimensional gel electrophoresis or two-dimensional-high-performance liquid chromatography (2 D-HPLC). The aim of this work was to establish a fully automated on-line 2-D-HPLC separation method with column switching for the separation of complex tryptic digests. A model mixture of five proteins as well as a nuclear matrix protein sample were digested with trypsin and separated using a strong cation exchange (SCX) column in the first dimension and nano reversed phase in the second dimension. Separated peptides were detected using an ion trap mass spectrometer. The advantages of this new fully automated method are rapid sample loading, the possibility of injecting large volumes and no introduction of salt into the mass spectrometer. Furthermore, column switching allows the independent control and optimization of the two dimensions independently. PMID- 15352230 TI - Characterization of a digested protein complex with quantitative aspects: an approach based on accurate mass chromatographic analysis with Fourier transform ion cyclotron resonance mass spectrometry. AB - We describe a new approach for the characterization of a digested protein complex with quantitative aspects. Accurate masses of tryptic peptides in the digested complex were acquired by nano-liquid chromatography Fourier transform-ion cyclotron resonance mass spectrometry (MS). The conditions of the electrospray ion source were alternated to acquire normal and fragment-ion-rich mass spectra concurrently. This, alternating-scan method, which includes no tandem mass spectrometry (MS/MS), allowed us to retain the integrity of the mass chromatograms and averted missed peptides due to MS and MS/MS switching. Tentative assignments of accurate peptide masses were verified with the concurrently acquired fragment-ion-rich spectra, and the identities of the protein components were established. For each identified protein component, mass chromatograms attributable to the validated accurate peptide masses were extracted, and the peak areas of multiple mass chromatograms were standardized. The standardized peak areas appeared to reasonably reflect the molar ratio of the protein components in standard mixtures. This new approach was successfully applied to the characterization of a cyanobacterial photosystem II complex preparation. A clear difference in the standardized peak areas was observed between the two groups of identified components, namely eight stoichiometric photosystem II proteins and two minor copurified phycobiliproteins. PMID- 15352231 TI - Two-dimensional electrophoresis for the isolation of integral membrane proteins and mass spectrometric identification. AB - Acrylamide concentration, urea content, and the trailing ion used for sodium dodecyl sulfate (SDS)-gels modify electrophoretic protein mobilities in a protein dependent way. Varying these parameters we coupled two SDS-gels to a two dimensional (2-D) electrophoresis system. Protein spots in 2-D gels are dispersed around a diagonal. Hydrophobic proteins are well separated from water-soluble proteins which is the essential advantage of the novel technique. Mass spectrometric identification of previously unaccessible hydrophobic proteins is now possible. PMID- 15352232 TI - Analysis of the humoral immune response to immunoselected phage-displayed peptides by a microarray-based method. AB - We describe a novel approach for high-throughput analysis of the immune response in cancer patients using phage-based microarray technology. The recombinant phages used for fabricating phage arrays were initially selected via the use of random peptide phage libraries and breast cancer patient serum antibodies. The peptides displayed by the phages retained their ability to be recognized by serum antibodies after immobilization. The recombinant phage microarrays were screened against either breast cancer or healthy donor serum antibodies. A model-based statistical method is proposed to estimate significant differences in serum antibody reactivity between patients and normals. A significant tumor effect was found with most of the selected phage-displayed peptides, suggesting that recombinant phage microarrays can serve as a tool in monitoring humoral responses towards phage-displayed peptides. PMID- 15352233 TI - Database-independent, database-dependent, and extended interpretation of peptide mass spectra in VEMS V2.0. AB - The Virtual Expert Mass Spectrometrist (VEMS) program package was developed for flexible, automated, and manual de novo tandem mass spectrometry (MS/MS) protein sequencing, and includes accessory programs for matrix-assisted laser desorption/ionization-mass spectrometry (MS) interpretation, and generation of protein and peptide databases. VEMS V2.0 has been developed into a fast tool for combining database-independent and -dependent protein assignments in an extended analysis of MS/MS-peptide data. MS or MS/MS data can be directly recalibrated after the first search by fitting the data to the best search result using polynomial equations. The score function is an improvement of known scoring algorithms and can be adapted for any MS instrument type. In addition, VEMS offers a novel statistical model for evaluating the significance of the protein assignment. The novel features are illustrated by the analysis of the fragmentation spectra obtained by liquid chromatrography-MS/MS analysis of peptides from an anionic peroxidase enriched protein fraction from potato root tissue. The extended analysis mode resulted in the additional assignment of spectra for nine modified tryptic peptides and nine miscleaved peptides, in addition to the 45 spectra from regular tryptic peptides. Of the nine modified peptides, three were glycosylated. PMID- 15352234 TI - Automated methods for improved protein identification by peptide mass fingerprinting. AB - In order to maximize protein identification by peptide mass fingerprinting noise peaks must be removed from spectra and recalibration is often required. The preprocessing of the spectra before database searching is essential but is time consuming. Nevertheless, the optimal database search parameters often vary over a batch of samples. For high-throughput protein identification, these factors should be set automatically, with no or little human intervention. In the present work automated batch filtering and recalibration using a statistical filter is described. The filter is combined with multiple data searches that are performed automatically. We show that, using several hundred protein digests, protein identification rates could be more than doubled, compared to standard database searching. Furthermore, automated large-scale in-gel digestion of proteins with endoproteinase LysC, and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis, followed by subsequent trypsin digestion and MALDI TOF analysis were performed. Several proteins could be identified only after digestion with one of the enzymes, and some less significant protein identifications were confirmed after digestion with the other enzyme. The results indicate that identification of especially small and low-abundance proteins could be significantly improved after sequential digestions with two enzymes. PMID- 15352235 TI - Determination of protein-protein interactions through aldehyde-dextran intermolecular cross-linking. AB - A very simple strategy, based on the intermolecular cross-linking of associated proteins by using aldehyde-dextrans, has been proposed to detect protein-protein interactions. Aldehyde-dextran was able to cross-link different enzymes composed of several polypeptide chains (e.g., trypsin and penicillin G acylase), proteolyzated proteins (e.g., extracts from porcine pancreas) and finally, an immunocomplex (horseradish peroxidase/anti-horseradish peroxidase). This cross linked immunocomplex could be selectively adsorbed on immobilized anti-rabbit IgG. The presence of unspecific covalent attachment between unrelated protein molecules was not detected. Thus, this strategy permits the cross-linking of different protein components and avoids the formation of nonspecific protein protein associations. PMID- 15352236 TI - Proteome analysis of human mesothelial cells during epithelial to mesenchymal transitions induced by shed menstrual effluent. AB - Peritoneal endometriosis is the result of ectopic implantation and growth of endometrium tissue that has been regurgitated into the abdominal cavity during menstruation. We have previously shown that menstrual effluent induces epithelial to mesenchymal transitions (EMT) in mesothelial cells, which results in cell retraction and exposure of submesothelial extracellular matrix. Since endometrial tissue preferentially adheres to the extracellular matrix, adhesion of endometrial tissue to the peritoneum is facilitated. The EMT were shown to be associated with differential expression and phosphorylation of mesothelial proteins. Using radiolabeling and proteomics we detected changes in protein expression and phosphorylation that occur in mesothelial cells during the EMT process. The identity of 73 proteins, which were obtained from 324 analyzed spots, was confirmed. The expression of 35 proteins involved in organization of the cytoskeleton, signal transduction, regulation of the redox state, and production of ATP, was altered during the EMT process. Four of the identified proteins were differentially phosphorylated: annexin-1, an actin-binding protein and a substrate for receptor tyrosine kinases; tropomyosin-alpha, a regulator of actin filament stability and cell shape; elongation factor 1 delta; ATP synthase beta-chain. In conclusion, factors from menstrual effluent induce specific changes in the expression and phosphorylation status of structural, regulatory and metabolic proteins relevant to the complex process of EMT in mesothelial cells. PMID- 15352237 TI - Use of surface-enhanced laser desorption/ionization -time of flight to explore bacterial proteomes. AB - Surface-enhanced laser desorption/ionization (SELDI)-time of flight is a recent technology that allows proteomic analysis with limited material requirements. This characteristic makes it a valuable technique for microbiologists handling problematic samples, such as low cell number cultures. We compared three simple procedures for protein extraction from bacteria for compatibility with the ProteinChip Array; we also determined the amount of protein required for each analysis. The protocol for the SELDI analysis was evaluated by generating protein expression profiles of a Streptococcus pneumoniae strain grown in different conditions and those of different strains of the same species. The protocol also was successfully applied to a wide range of Gram positive and negative bacteria. The results of this study suggest the appropriateness of this technology for microorganism protein profiling as complementary or alternative to two dimensional gel electrophoresis. PMID- 15352238 TI - The identification of Dictyostelium phosphoproteins altered in response to the activation of RasG. AB - Dictyostelium RasG has been implicated in the regulation of a variety of cellular processes, including the initiation of development, cell movement, and cytokinesis, but the molecular components of the signaling pathways involved are largely unknown. We used a tetracycline-regulated protein expression system to study the effect of activated RasG, RasG(G12T), expression on the phosphorylation state of Dictyostelium proteins. Over 70 vegetative phosphoprotein components were resolved by two-dimensional (2-D) immunoblot analysis and of these 16 phosphothreonine and three phosphotyrosine protein components were found to reproducibly change upon RasG(G12T) expression. Thirteen of these were recovered from 2-D gels and identified by mass spectrometry of in-gel tryptic digestions. The proteins identified include the signaling proteins RasGEF-R and protein kinase B, the adhesion protein DdCAD-1, the cytoskeletal protein actin, the mitochondrial division protein FtsZA, and proteins involved in translation and metabolism. In addition to the direct demonstration of the phosphorylation of putative downstream targets of RasG activation, these findings reveal previously undetected phosphorylation of several proteins. PMID- 15352239 TI - Insights into Pseudomonas putida KT2440 response to phenol-induced stress by quantitative proteomics. AB - To gain insight into the global mechanism underlying phenol toxicity and tolerance in bacteria, we have generated a two-dimensional protein reference map and used it to identify variations in protein expression levels in Pseudomonas putida KT2440 following exposure to sub-lethal inhibitory concentrations of this solvent. Inspection of the two-dimensional gel electrophoresis gels revealed that 1 h following sudden cell exposure to two different concentrations of phenol, leading to the inhibition of exponential growth (600 mg/L) or to growth arrest for, at least, 4 h before inhibited growth resumption (800 mg/L), the amount of 68 proteins was increased while the amount of 13 proteins was reduced. The up regulated proteins include proteins involved in the: (i) oxidative stress response (AhpC, SodB,Tpx and Dsb); (ii) general stress reponse (UspA, HtpG, GrpE and Tig); (iii) energetic metabolism (AcnB, AtpH, Fpr, AceA, NuoE, and MmsA-1); (iv) fatty acid biosynthesis (FabB, AccC-1 and FabBx1); (v) inhibition of cell division (MinD); (vi) cell envelope biosynthesis (LpxC, VacJ, and MurA); (vii) transcription regulation (OmpR and Fur); and (viii) transport of small molecules (TolC, BraC, AotJ, AapJ, FbpA and OprQ). Among the down-regulated proteins are those involved in nucleotide biosynthesis (PurM, PurL, PyrH and Dcd) and cell motility (FliC). The information emerging from this genome expression profiling and the detailed investigation of the biological role of candidate genes, as targets of phenol toxicity or as determinants of phenol resistance in P. putida KT2440, will allow more rationale strategies for developing bacteria with greater solvent tolerance with impact in bioremediation and whole-cell biotransformations in media with organic solvents. PMID- 15352240 TI - Identification of Bacillus anthracis proteins associated with germination and early outgrowth by proteomic profiling of anthrax spores. AB - The use of anthrax spores as a bioweapon has spurred efforts aimed at identifying key proteins expressed in Bacillus anthracis. Because spore germination and outgrowth occur prior to and are required for disease manifestations, blocking germination and early outgrowth with novel vaccines or inhibitors targeting critical B. anthracis germination and outgrowth-associated factors is a promising strategy in mitigating bioterror. By screening 587 paired protein spots that were isolated from dormant and germinating anthrax spores, respectively, we identified 10 spore proteins with statistically significant germination-associated increases and decreases. It is likely that proteins whose levels change during germination may play key roles in the germination and outgrowth processes, and they should be listed as priority targets for development of prophylactic and therapeutic agents against anthrax. The 31 new proteins identified in this study also complement an emerging proteomic database of B. anthracis. PMID- 15352241 TI - Protein composition of Paracoccus denitrificans cells grown on various electron acceptors and in the presence of azide. AB - Two-dimensional gel electrophoresis (2-DE) with immobilized pH gradients was carried out on total cell lysates and membrane fractions of Paracoccus denitrificans with the aim to characterize differences in protein expression during growth under aerobic and various anaerobic conditions (with nitrate, nitrite or nitrous oxide). Comparative image analysis of the protein pattern revealed several subgroups of the total 800 protein spots resolved that were characteristically induced or repressed in response to individual electron acceptors. The respiratory inhibitor azide also exerted a profound influence upon cellular protein composition. However, since most of the proteins showing an altered expression pattern in cells growing on oxygen differed from those in cells growing on nitrite, we suppose that azide acts mainly indirectly, possibly by influencing other cellular signals. Limited information on the P. denitrificans genome has precluded the identification of more than eight protein spots as yet. A public accessible P. denitrificans 2-DE protein database is currently built up at http://www.mpiib-berlin.mpg.de/2D-PAGE. PMID- 15352242 TI - Wheat leaf proteome analysis using sequence data of proteins separated by two dimensional electrophoresis. AB - Identifying wheat leaf protein expression is a major challenge of functional genomics. Using two-dimensional gel electrophoresis 541 wheat leaf proteins were separated and 55 of them were sequenced by nano liquid chromatography-tandem mass spectrometry. Peptide sequence data were screened against protein banks and expressed sequence tag public banks. Among these 55 spots, 20 proteins were found in wheat and 21 in other grass families (http://www.ncbi.nlm.nih.gov/). Twelve proteins showed similarities with other eukaryotic plant species. One protein showed homology to a bacterial sequence and another protein remained unknown. In 18 cases a significant score was found for the wheat TUC (Tentative Unique Contigs) of the PlantGDB (http://www.plantgdb.org/) data. In several cases, different spots were identified as corresponding to the same protein that can probably be attributed to the hexaploid structure of wheat. The identified proteins were classified in six groups and their role is discussed. Most of them (31/55) are involved in carbohydrate metabolism. PMID- 15352243 TI - Proteomic analysis of aneuploid lines in the homeologous group 1 of the hexaploid wheat cultivar Courtot. AB - Three monosomic lines (MSLs) and three nullisomic lines (NSLs) of the homeologous group 1 and one euploid line of the bread wheat Triticum aestivum cultivar Courtot were used in a proteomic approach to investigate the effects of zero, one or two doses of chromosomes 1A, 1B and 1D on the amount of endosperm proteins. Polypeptides whose amounts changed significantly between each aneuploid line and the euploid line were identified using image analyses of two-dimensional gel electrophoresis patterns resulting from specific endosperm protein extractions. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry and electrospray ionization tandem mass spectrometry were also used for protein identification. Removing one chromosome or a chromosome pair allowed varying responses to be observed for the remaining endosperm protein genes. Compensation phenomena for the high molecular weight glutenin subunits (HMW-GS) were detected only in the MSLs. Subunits Bx7, By8 and Dy12 were the only HMW-GS overexpressed (from 152-737%) when chromosomes 1A or 1B or 1D were at hemizygous state. Thirteen new protein spots were detected only in the NSL1D, and seven were identified as HMW-GS analogs. These seven new spots may result from the expression of inactive genes. The HMW-GS were of significantly higher volume in MSLs, whereas the low molecular weight glutenin subunits and the gamma-gliadins were of lower volume in aneuploid lines. Most of the down-regulated proteins in the MSLs were storage proteins encoded at loci located on another chromosome pair. Complex regulations between chromosomes and loci of the homeologous groups 1 and 6 in bread wheat are discussed. PMID- 15352244 TI - New targets of Arabidopsis thioredoxins revealed by proteomic analysis. AB - Proteomics was used to search for putative thioredoxin (TRX) targets in leaves of the model plant, Arabidopsis thaliana. About forty different proteins have been found to be reduced by TRX, after TRX itself has been specifically reduced by its NADPH-dependent reductase. Twenty-one of the identified proteins were already known or recently proposed to be TRX-dependent and nineteen of the proteins were new potential targets. The identified proteins are involved in a wide variety of processes, including the Calvin cycle, metabolism, photosynthesis, folding, defense against oxidative stress and amino acid synthesis. Two proteins from the glycine cleavage complex were also identified as putative TRX targets, and a new role can be postulated in leaves for TRX in defense against herbivores and/or pathogens. PMID- 15352245 TI - Homology-based functional proteomics by mass spectrometry: application to the Xenopus microtubule-associated proteome. AB - The application of functional proteomics to important model organisms with unsequenced genomes is restricted because of the limited ability to identify proteins by conventional mass spectrometry (MS) methods. Here we applied MS and sequence-similarity database searching strategies to characterize the Xenopus laevis microtubule-associated proteome. We identified over 40 unique, and many novel, microtubule-bound proteins, as well as two macromolecular protein complexes involved in protein translation. This finding was corroborated by electron microscopy showing the presence of ribosomes on spindles assembled from frog egg extracts. Taken together, these results suggest that protein translation occurs on the spindle during meiosis in the Xenopus oocyte. These findings were made possible due to the application of sequence-similarity methods, which extended mass spectrometric protein identification capabilities by 2-fold compared to conventional methods. PMID- 15352246 TI - Surface-enhanced laser desorption/ionization-time of flight-mass spectrometry approach to biomarker discovery in blue mussels (Mytilus edulis) exposed to polyaromatic hydrocarbons and heavy metals under field conditions. AB - Proteomics provide potential in the discovery of new sensitive biomarkers for environmental pollution. To evaluate this potential, we have utilized ProteinChip technology to analyze the proteomic profile of blue mussels (Mytilus edulis) from polluted marine habitats surrounding the island of Karmoy, Norway. Two different types of contamination, heavy metals and polyaromatic hydrocarbons (PAHs), were compared to a clean reference site. Differentially expressed proteins/peptides were found, which showed a specific induction or a general suppression associated with the field site of origin. By combining sets of protein markers in a tree building algorithm, we were able to correctly classify samples from these sites with an accuracy of 90%. PMID- 15352247 TI - Identification of protein targets for mycophenolic acid acyl glucuronide in rat liver and colon tissue. AB - Covalent binding of acyl glucuronides to proteins is considered an initiating event for the organ toxicity of drugs containing a carboxylic acid group. An acyl glucuronide (AcMPAG) of the immunosuppressant mycophenolic acid was described and shown to form covalent adducts with plasma albumin in vivo. The aim of the present investigation was to identify AcMPAG target proteins in the liver and colon of rats treated with mycophenolate mofetil, which may contribute to a better understanding of the mechanisms responsible for the development of side effects during therapy with this drug. Mycophenolate mofetil was administered per os in to Wistar rats (40 mg/kg/day) over 21 days. Proteins in liver and colon homogenates were separated by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. AcMPAG labeled protein spots were detected by Western blotting. After in-gel tryptic digestion of the protein spots from parallel gels (n = 2), peptides were characterized by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. Data base searching identified AcMPAG target proteins. Tryptic peptides with sufficient signal intensities were subjected to post-source decay analysis. Three proteins in the liver (ATPase/ATP synthase (alpha and beta subunits), protein disulfide isomerase A3 and selenium binding protein) and one protein in the colon (selenium binding protein) were identified as targets for AcMPAG. ATPase/ATP synthase and protein disulfide isomerase are essential proteins involved in the control of the energy and redox state of the cells, whereas the physiological role of selenium binding protein is not fully understood. This study shows for the first time the formation of adducts between tissue proteins and AcMPAG. Whether this chemical modification is associated with compromised protein function and drug toxicity remains to be investigated. PMID- 15352248 TI - Evaluation of an integrated strategy for proteomic profiling of skeletal muscle. AB - Proteomic analysis of skeletal muscle presents particular challenges when trying to identify valid biomarkers of phenotypic change in small biopsies from genetically diverse human subjects. Currently, two-dimensional (2-D) gel electrophoresis and mass spectrometry are the chosen analytical strategies but 2 D gels are not appropriate for analyzing proteins less than 11 kDa, they can suffer from problems of reproducibility and in routine use are not a viable high throughput technique. We have evaluated an integrated proteomic strategy employing Ciphergen ProteinChip arrays, one-dimensional polyacrylamide gel electrophoresis and mass spectrometry. Protein fingerprints characteristic of fast and slow contracting muscles from normal and kyphoscoliosis (ky) mutant mice were obtained from Ciphergen protein arrays. Eight statistically validated protein biomarkers have so far been identified capable of discriminating fast from slow muscle. Five of these showed further differential expression in ky versus normal BDL soleus muscles. Several biomarkers have been formally identified, and were myosin light chain isoforms shown previously to be expressed differentially by fast versus slow skeletal muscles. This integrated experimental approach using a model mouse muscle system shows the potential of Ciphergen protein array technology for proteomic analysis of small proteins in small muscle samples and its applicability for phenotypic characterization of skeletal muscle in general. PMID- 15352249 TI - Identification of metastasis-associated proteins in a human tumor metastasis model using the mass-mapping technique. AB - For most cancer cell types, the acquisition of metastatic ability leads to clinically incurable disease. The identification of molecules whose expression is specifically correlated with the metastatic spread of cancer would facilitate the design of therapeutic interventions to inhibit this lethal process. In order to facilitate metastasis gene discovery we have previously characterized a pair of monoclonal cell lines from the human breast carcinoma cell line MDA-MB-435 that have different metastatic phenotypes in immune-compromised mice. In this study, serum-free conditioned media was collected from the cultured monoclonal cell lines and a mass mapping technique was applied in order to profile a component of each cell line proteome. We utilized chromatofocusing in the first dimension to obtain a high resolution separation based on protein pI, and nonporous silica reverse-phase high performance liquid chromatography was used for the second dimension. Selected proteins were identified on the basis of electrospray ionization time of flight mass spectrometry (ESI-TOF MS) intact protein mapping and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) peptide mass fingerprinting. Using this approach we were able to map over 400 proteins and plot them as a 2-D map of pI versus accurate M(r). This was performed over a pI range of 4.0-6.2, and a mass range of 6-80 kDa. ESI-TOF MS data and further analysis using MALDI-TOF MS confirmed and identified 27 differentially expressed proteins. Proteins associated with the metastatic phenotype included osteopontin and extracellular matrix protein 1, whereas the matrix metalloproteinase-1 and annexin 1 proteins were associated with the non metastatic phenotype. These findings demonstrate that the mass mapping technique is a powerful tool for the detection and identification of proteins in complex biological samples and which are specifically associated with a cellular phenotype. PMID- 15352250 TI - Identification of tumor-associated plasma biomarkers using proteomic techniques: from mouse to human. AB - In an effort to identify tumor-associated proteins from plasma of tumor-bearing mice that may be used as diagnostic biomarkers, we developed a strategy that combines a tumor xenotransplantation model in nude mice with comparative proteomic technology. Five human cancer cell lines (SC-M1, HONE-1, CC-M1, OECM1, GBM 8401) derived from stomach, nasopharyngeal, colon, oral and brain cancers were subcutaneously inoculated into nude mice and compared to control nude mice injected with phosphate-buffered saline. One month later, plasma from mice inoculated with cancer cells was collected for proteomic analysis using two dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). Comparison of plasma 2-DE maps from tumor-bearing mice with those produced from control mice revealed the overexpression of several mouse acute phase proteins (APPs) such as haptoglobin. Another APP, serum amyloid A (SAA), was found only in mice bearing tumors induced by the stomach cancer cell line SC-M1, which has not previously been demonstrated in xenotransplatation experiment. Furthermore, by using immunohistochemistry, SAA and haptoglobin were found to originate from the mouse hosts and not from the human cancer cell line donors. The protein alterations were further confirmed on patients with stomach cancers where up-regulated levels of SAA were also observed. These results indicate that APPs may be used as nonspecific tumor-associated serum markers. SAA in particular may serve as a potential marker for detecting stomach cancer. Taken together, the combination of the xenotransplatation model in nude mice and proteomics analysis provided a valuable impact for clinical applications in cancer diagnostics. In addition, our findings demonstrate that a panel of APPs might serve as screening biomarkers for early cancer detection. PMID- 15352251 TI - Proteomic signature of human cancer cells. AB - We assessed proteomic profiles as biomarkers for monitoring cell phenotypes. Protein expression profiles were obtained by fluorescence two-dimensional difference gel electrophoresis (2-D-DIGE), in which quantitative ability is improved by labeling proteins with fluorescent dyes prior to electrophoresis. Integrated protein spot intensities were analyzed by a statistical approach. The proteomic data of two groups of cell lines: (1) adenocarcinoma (AC) cell lines derived from lung, pancreas and colon tissues and (2) lung cancer cell lines with different histological backgrounds, including AC, squamous cell carcinoma and small cell carcinoma, were assessed on the basis of prior biological information. Hierarchical clustering analysis and principal component analysis were used to divide the cell lines into subgroups on the basis of similarities between their protein expression profiles. The majority of cell lines were grouped according to their organ of origin or histological background. A machine-learning algorithm selected 32 protein spots that were responsible for the classification. The results indicate that proteomic data generated by 2-D-DIGE can provide a signature of essential cell phenotypes, suggesting that it might be possible to apply this technique to developing tumor markers that could identify the organ of origin of metastatic tumors and contribute to the differential diagnosis of lung cancer. PMID- 15352252 TI - Mitochondrial proteome: cancer-altered metabolism associated with cytochrome c oxidase subunit level variation. AB - Shifts in metabolism associated with tumorigenesis were first noted by Otto Warburg in the 1920s. In the ensuing decades many examples of the phenomenon have been elucidated while the underlying molecular mechanism has remained elusive. As the enzyme complex at the crux of oxidative phosphorylation, cytochrome c oxidase is uniquely positioned to have a very high impact on cellular metabolism. In this study, we test the hypothesis that there is a specific association between altered cytochrome c oxidase subunit levels and altered metabolism by combining the technique of reverse-phase protein microarray with radiolabeled glucose metabolic studies. Such a relationship is observed with five different cell lines, two of which (1542N and 1542T) are a matched set of normal and tumor-based lineages derived from the same prostate gland. By measuring the [(14)C]carbon dioxide production of a cell line metabolizing [1-(14)C]glucose and comparing those measurements to values obtained for the same cell line metabolizing [6 (14)C]glucose, we determined the relative utilization of the hexose monophosphate shunt and glycolysis progressing through the Krebs cycle metabolic pathway in each cell line. In all cases there is an increased utilization of hexose monophosphate shunt relative to glycolysis progressing through the Krebs cycle in tumor derived relative to normal derived cell lines. Additionally, there is an associated increase in the ratio of nuclear encoded cytochrome c oxidase subunits to mitochondrially encoded cytochrome c oxidase subunits in the tumor-derived cell lines. These results demonstrate an alteration in subunit levels of a single enzyme complex (cytochrome c oxidase) commensurate with tumor-altered metabolism. PMID- 15352253 TI - Profiling treatment-specific post-translational modifications in a complex proteome with subtractive substrate phage display. AB - Proteolytic activation of zymogens or controlled degradation of inhibitory factors is part of a major regulatory system on the post-translational level to regulate treatment induced cellular stress responses. The identification of differential activity based substrates is thus of high interest to prioritize and validate candidate targets for drug discovery. Here we present a novel subtractive substrate phage display screening method for the selection of treatment induced post-translational peptide modifications in complex proteomes. We investigated this approach with tumor cells in response to a protease activating anticancer treatment modality using subtractive and iterative screening of cellular extracts derived from control and treated cells. Specific phage were identified that served as substrates for proteolytic activities in response to treatment related activity changes and could be distinguished from substrates for unspecific proteolytic background activities. Novel, selected peptide substrates were investigated in vitro and in vivo and showed high substrate specificity and functional biological significance. PMID- 15352254 TI - Proteomic analysis of differential protein expression in neuropathic pain and electroacupuncture treatment models. AB - Acupuncture has long been used for pain relief. Although recent studies have shown that acupuncture can reduce neuropathic pain, the mechanism of this effect is not clear and little information is available regarding proteins that are involved in the development of neuropathic pain and the effects of acupuncture. We have developed an animal model for neuropathic pain using young adult male Sprague-Dawley rats. The model was confirmed by behavioral tests. Electroacupuncture (EA) treatment was applied to Zusanli (ST36) of neuropathic pain model to examine the analgesic effect of EA. The protein expression profile of the hypothalamus in both neuropathic pain and EA treatment models was analyzed using two-dimensional electrophoresis-based proteomics. We detected thirty-six proteins that were differentially expressed in the neuropathic pain model compared with normal rats and that restored to normal expression levels after EA treatment. Twenty-one of these proteins were identified in the MS-FiT database and are involved in a number of biological processes, including inflammation, enzyme metabolism and signal transduction. Potential applications of our results include the identification and characterization of signaling pathways involved in EA treatment and further exploration of the role of selected identified proteins in the animal model. PMID- 15352255 TI - Proteomic analysis of proteins altered by dibenzoylmethane in human prostatic cancer LNCaP cells. AB - This paper explores the use of proteomics as a tool for identifying protein species whose expression has been altered by dibenzoylmethane (DBM) in LNCaP cells. Although DBM, a constituent of licorice, has been shown to induce cell cycle arrest and regulate androgen receptor (AR) expression, the mechanism by which these events occur is unknown. To develop a better understanding of the effect of DBM on cancer cells, we analyzed changes in protein expression induced by DBM in LNCaP cells using two-dimensional (2-D) gel electrophoresis. The proteomic approach used to study LNCaP cells has lead to the analysis and identification of a number of protein species that increase or decrease as a result of exposure to DBM. In particular, twenty features were found to be differentially expressed in this study based on the quantitation of two separate 2-D-fluorescence difference gel electrophoresis analyses. Thirteen of these features were identified through mass spectrometric analysis. The intensity of 10 out of the 13 spots identified increased 2- to 3-fold in response to 25 micro M and 50 micro M DBM and the remaining three spots decreased 2-fold in response to the same DBM treatment. This study investigates proteomic changes induced by treatment of cells with DBM in order to develop a model for the mechanism by which DBM induces cell cycle arrest and represses AR expression. PMID- 15352261 TI - Toxicity of the cyanobacterium Cylindrospermopsis raciborskii to Daphnia magna. AB - The effect of two strains of Cylindrospermopsis raciborskii on the survivorship, somatic growth, and detoxification processes of juvenile Daphnia magna were investigated. Both strains of C. raciborskii (and also Ankistrodesmus falcatus, used as the control) were given to newborn D. magna at equivalent biovolumes. The survival curves for D. magna subjected to the two C. raciborskii treatments differed from those of the starved and fed treatments. After 48 h of exposure, the percentage of D. magna surviving after exposure to Cylin-A (a cylindrospermopsin-producing strain isolated from Australia) and Cylin-P (a non cylindrospermopsin-producing strain isolated from Portugal) was 10.00% and 93.33%, respectively. The strain that produces cylindrospermopsin caused the greatest toxic effect in juvenile D. magna. Statistically significant differences in D. magna body size between the four treatments (Cylin-A, Cylin-P, A. falcatus, and starved) were detected after 48 h of exposure. The juvenile D. magna that received the two C. raciborskii treatments showed an increase in size (relative to their size at T(0)) of 2.54% and 38.14%, respectively. These values were statistically significantly different than those of the A. falcatus-fed control (55.54%) and the starved control (11.47%). In both C. raciborskii treatments there was a tendency for increased GST enzyme activities after 24 h of exposure. Cylindrospermopsin was detected (HPLC-MS/MS) in D. magna tissues after 24 and 48 h (0.025 and 0.02 ng animal(-)1, respectively). The results of this study indicate that C. raciborskii can affect the fitness and growth potential of juvenile D. magna. PMID- 15352262 TI - Xenobiotic substances such as PCB mixtures (Aroclor 1254) and TBT can influence swimming behavior and biotransformation activity (GST) of carp (Cyprinus carpio). AB - Different groups of carp were treated with polychlorinated biphenyl (PCB) or tributyltin (TBT), and possible effects of the chemicals on the swimming behavior of the carp were examined using the BehavioQuant system. By evaluating quantitative behavioral parameters of the animals, it became evident that exposure to high concentrations of chemicals (organotin, 7 microg L(-1), or polychlorinated biphenyl, 22 microg L(-1)) severely affected the carp, causing a significant change in their swimming speed. TBT stress led to a rapid decrease in mean swimming activity. A decrease in the preferred swimming depth was observed in TBT- and PCB-exposed fish. Animals exposed to PCB reduced their mean daily activity and increased their mean swimming speed in the nighttime during the second week of exposure. Therefore, our findings imply that the fish were adapted to cope with the chemicals after the second week of exposure. Furthermore, our results showed that low concentrations (TBT, 0.3 and 2 microg L(-1), or PCB, 14 microg L(-1)) did not significantly alter any quantified parameters of swimming behavior. In addition, the direct effects of chemicals on enzyme activity (GST) were determined. Measurement of soluble glutathione-S-transferase activity of fish liver or gills showed a significant elevation after exposure to PCB (0.1 or 22 microg L(-1)) or TBT (0.8 or 7 microg L(-1)). We had to conclude that the two different end points tested generally are useful as biomarkers of exposure and for investigations of energy resources in organisms under the influence of toxic stress. PMID- 15352263 TI - Evaluating acute toxicity of methyl parathion application in constructed wetland mesocosms. AB - Wetland ecosystems have reduced ambient levels of various organic and metallic compounds, although their effectiveness on agricultural pesticides is not well documented. Five stations within each of two 10 x 50 m constructed wetlands (two vegetated, two nonvegetated) were selected to measure the fate and effects of methyl parathion (MeP). Following a simulated storm event (0.64 cm of rainfall), aqueous, sediment, and plant samples were collected and analyzed spatially (5, 10, 20, and 40 m from the inlet) and temporally (after 3-10 days) for MeP concentrations and for the impact of those concentrations on the aquatic fauna. Aqueous toxicity to fish decreased spatially and temporally in the vegetated mesocosm. Pimephales promelas survival was significantly reduced, to 68%, at the 10-m station of the nonvegetated wetlands (3 h postapplication), with pesticide concentrations averaging 9.6 microg MeP/L. Ceriodaphnia in both the vegetated and nonvegetated wetlands was sensitive (i.e., a significant acute response to MeP occurred) to pesticide concentrations through 10 days postapplication. Mean MeP concentrations in water ranged from 0.5 to 15.4 microg/L and from 0.1 to 27.0 microg/L in the vegetated and nonvegetated wetlands, respectively. Hyalella azteca aqueous tests resulted in significant mortality in the 5-m vegetated segment 10 days after exposure to MeP (2.2 microg/L). Solid-phase (10-day) sediment toxicity tests showed no significant reduction in Chironomus tentans survival or growth, except for the sediments sampled 3 h postapplication in the nonvegetated wetland (65% survival). Thereafter, midge survival averaged >87% in sediments sampled from both wetlands. These data suggest that wetlands play a significant role in mitigating the effect of MeP exposure in sensitive aquatic biota. PMID- 15352264 TI - Interaction between halogenated aromatic compounds in the Ah receptor signal transduction pathway. AB - Many toxic and biochemical responses to halogenated aromatic compounds (HACs) such as polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins (PCDDs) are mediated through the aryl hydrocarbon receptor (AhR), which is an intracellular cytosolic target for HACs. Environmental exposure to HACs almost always involves complex mixtures of congeners, some of which can antagonize the action of potent HACs such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In this work we studied TCDD and representative PCB congeners, alone and in mixture, for their effect on CYP1A gene transcription and protein levels in primary rat hepatocytes. Together with our previous work, our results suggest that formation of the Ah receptor-ligand-DRE (dioxin response element) complex is the principal point of divergence in the mechanism between an AhR agonist and an AhR antagonist. The coplanar PCBs 77 and 126 and the mono-ortho PCB 156 were full agonists toward CYP1A1 gene transcription and CYP1A protein levels, showing typical additive behavior with TCDD to the target molecule AhR. In contrast, the nonplanar PCB 153 antagonized the action of TCDD, even at concentrations that occupied a significant fraction of AhR molecules. Competitive inhibition explains the commonly reported decrease of ethoxyresorufin-O-deethylase (EROD) activity when PCBs are present in high concentrations and the antagonism of PCBs to the EROD activity of TCDD. The result is that Western blotting offers a much more reliable measure of CYP1A protein concentration than does the EROD assay, despite the greater convenience of the latter. PMID- 15352265 TI - Assessing cytotoxicity of photosensitized transformation products of 2,4,6 trinitrotoluene (TNT) and atrazine with freshwater microbial assemblages. AB - In this study of riboflavin-sensitized photolysis of 2,4,6-trinitrotoluene (TNT), the final photoproducts were found to include 2-amino-4,6-dinitrotoluene, 4-amino 2,6-dinitrotoluene, and 3,5-dinitroaniline. After exposure to TNT (10 mg/L) for 90 min, in a river water sample there was inhibition of the viability count of heterotrophic bacterial assemblages by 28.3% and 24.3% and of bacterial heterotrophic mineralization of glucose by 99.5% and 93.6% relative to the light control and dark control groups, respectively. After exposure separately to 10 mg/L 2-amino-4,6-dinitrotoluene, 4-amino-2,6-dinitrotoluene, or 3,5 dinitroaniline, the viability count of heterotrophic bacterial assemblages was enhanced by 30.0%, 98.2%, and 148.7%, respectively, relative to the TNT group in light and by 12.2%, 40.2%, and 36.5%, respectively, relative to TNT group in dark. After exposure to the same aforementioned test chemicals, heterotrophic activity of bacterial assemblages was inhibited by 75.7%, 72.4%, and 56.0%, respectively, relative to light control group, and there were no significant changes in the dark. A similar study was conducted with atrazine (10 mg/L). The main products of atrazine riboflavin-sensitized phototransformation include desethylatrazine (DEA) and desisopropylatrazine (DIA). It was found that DEA and DIA did not inhibit microbial heterotrophic activity. However, after 72 h the viable count was enhanced by 52.8% and 63.3% in the DEA and DIA exposure groups, respectively, in comparison to the control. These results suggested that photosensitized transformation decreased TNT and atrazine cytotoxicity to microbial assemblages in the natural water. Photoproducts of atrazine--DEA and DIA, and of TNT could become growth substrates for bacterial assemblages in natural water. PMID- 15352266 TI - Tropical ecotoxicity testing with Ceriodaphnia cornuta. AB - The Sai Gon-Dong Nai river system in southern Vietnam is of great social and economic importance yet receives a large amount of industrial, domestic, and agricultural discharges. Toxicity assessment has started to become an issue in Vietnam, and it is important to employ a test system that is appropriate for typical Vietnamese conditions with a species that is representative of the invertebrates living in its aquatic ecosystems. The aim of this study was to develop and to validate an ecotoxicity test with an autochthonous organism. The microcrustacean Ceriodaphnia cornuta (Cladocera) was isolated from the Sai Gon River. A protocol was developed for the culturing of the organisms, and satisfactory results were obtained for long-term reproduction of C. cornuta. Quality control criteria were established. The toxicity of two relevant pesticides (diazinon and methyl parathion) and two metals (chromium and mercury) to C. cornuta was evaluated and compared to the standard organisms for ecotoxicological testing: Daphnia magna and Vibrio fischeri. Nonlinear regression models were applied to estimate such parameters as EC(50). The results of acute toxicity tests showed that C. cornuta was less tolerant than D. magna and V. fischeri to salinity and that C. cornuta was more sensitive than D. magna and V. fischeri to potassium dichromate, diazinon, methyl parathion, and mercury. PMID- 15352267 TI - Circulating immune complexes as indicators of environmental contamination. AB - The aim of this study was to examine the effect of environmental contamination on the concentration of circulating immune complexes (CIC) in bovine blood. We determined that in a conventionally ecologically clear district, the CIC concentration in healthy bovine blood serum was within the limits of 0.18-0.52 mg/mL, (mean = 0.31 mg/mL, V = 30.14%). In a conventionally ecologically contaminated district, the CIC concentration in healthy bovine blood serum was detected within the limits of 0.48-0.99 mg/mL (mean = 0.65 mg/mL, V = 17.97%; P < 0.05). The CIC concentration in serum of bovine leukemia virus (BLV)-infected bovines from different districts was within the limits of 0.29-1.76 mg/mL (mean = 0.81 mg/mL, V = 36.67%; P < 0.05). According to these preliminary results of the investigation, it may be supposed that the increased concentration of CIC is closely related to the environmental contamination. Therefore, studies of CIC concentration in healthy mammals are of great importance for the monitoring of anthropogenic factors and can be exploited as an indicator of environmental contamination. PMID- 15352268 TI - In vitro androgenicity in pulp and paper mill effluents. AB - Pulp and paper mill effluents were examined for in vitro androgenicity using a recombinant yeast-based androgen receptor assay. Low levels of androgenic effects were detected in extracts of some effluents after activated sludge treatment. Yeast cell growth cell was inhibited in some samples, and this interfered with the androgenic response. Solid-phase fractionation revealed androgenicity after partial separation of components in the effluents. Comparison of levels in untreated effluent and in effluent treated in an aerated lagoon showed that this treatment had only a marginal effect on androgenicity. An assay of the fractions eluted with increasing concentrations of methanol showed that androgenic compounds were low to moderately lipophilic. In an attempt to identify these compounds, a number of wood-related compounds (guaiacol, vanillin, beta sitosterol, betulin, pinosylvin-O-methyl ether, and a wood extract enriched in lignans) were examined but were found not to be androgenic. Raw process water was not androgenic, but water from a highly humified lake and process water from the production of pulp from partly decayed wood had low androgenicity. It therefore can be plausibly suggested that the androgens originated in decaying wood. An assay of androgenicity in the bile of juvenile rainbow trout exposed to effluents for 3 weeks showed increased dose-dependent levels of androgens after enzymatic hydrolysis of hormone conjugates. PMID- 15352269 TI - Life-history traits of standard and autochthonous cladocerans: I. Acute and chronic effects of acetylsalicylic acid. AB - Pharmaceuticals have been recognized as an important group of aquatic micropollutants, mainly because of their biologically active nature. Acetylsalicylic acid (ASA), which is the active compound of Aspirin and many other pharmaceuticals, is consumed in large quantities every year. Therefore, its acute and chronic effects on standard (Daphnia magna) and autochthonous (Daphnia longispina) daphnids were investigated. The results showed that ASA impaired the survivorship, reproduction, and growth of the cladoceran species. The standard daphnid was the more tolerant species in acute assays (48-h EC(50) = 1293.05 mg/L; D. longispina: 48-h EC(50) = 647.31 mg/L); whereas the autochthonous daphnid seemed to be more resistant under chronic exposure to ASA, mainly its population-level traits. Despite this, the observed effect concentrations were much higher than the environmental concentrations of ASA. Notwithstanding this, the impairment of individual-level traits is likely to occur at environmental levels as an ultimate response to long-term exposure. PMID- 15352270 TI - Life-history traits of standard and autochthonous cladocerans: II. Acute and chronic effects of acetylsalicylic acid metabolites. AB - Metabolic products are often more toxic than their pharmacological parent compounds. Therefore, the acute and chronic effects of the main metabolites- salicylic acid (SAL), gentisic acid (GEN), and o-hydroxyhippuric acid (HDP)--of acetylsalicylic acid (ASA), the active ingredient in Aspirin and many other pharmaceuticals, were assessed using standard (Daphnia magna) and autochthonous (Daphnia longispina) cladocerans. The sequence of decreasing levels of acute and chronic toxicity of ASA metabolites to daphnids was GEN > SAL > HDP. HDP did not present acute toxicity, but chronic exposures enabled the production of abnormal neonates and, in particular, egg abortion. Thus, reproduction was the end point most susceptible to HDP. On the other hand, SAL and GEN induced changes in the normal patterns of reproduction and growth of both species. In general, D. longispina was more sensitive than was D. magna, although the population growth of the autochthonous species was superior under SAL exposures than that of the standard test species. Although the concentrations that were determined to have a toxic effect were above the levels detected in aquatic environmental samples, exposure to low levels of pharmacologically active substances for a duration longer than the test period may induce changes in nontarget organisms. PMID- 15352271 TI - First report of cyanobacterium Cylindrospermopsis raciborskii from Algerian freshwaters. AB - This study investigated the first report of the cyanobacterium Cylindrospermopsis raciborskii in Algerian freshwaters. The morphological characteristics of the two morphotypes observed in Lake Oubeira (Algeria) conformed to those of natural populations of C. raciborskii species described in the literature. The two morphotypes produced only straight trichomes. During the study (February 2000 September 2001), this species occurred as a codominant with Microcystis spp., and a peak was observed during the autumn (November 2000) with a remarkable density estimated to be 43 x 10(5) trichomes/L. However, difficulty in isolating and culturing these two morphotypes limited the evaluation of their toxic potential. PMID- 15352272 TI - From clinical observations to clinimetrics: a tribute to Alvan R. Feinstein, MD. PMID- 15352273 TI - Behavioral Neurobiology of Birdsong. Proceedings of a conference. New York, New York, USA, 12-14 December 2002. PMID- 15352274 TI - Homocysteine as a predictive factor for hip fracture in older persons. PMID- 15352275 TI - Homocysteine as a predictive factor for hip fracture in older persons. PMID- 15352276 TI - Homocysteine as a predictive factor for hip fracture in older persons. PMID- 15352277 TI - Myostatin mutation associated with gross muscle hypertrophy in a child. PMID- 15352278 TI - Case 18-2004: a 61-year-old man with rectal bleeding. PMID- 15352279 TI - WALTHAM International Science Symposium: Nature, Nurture, and the Case for Nutrition. Proceedings of a conference. October 28-31, 2003, Bangkok, Thailand. PMID- 15352283 TI - Steroid drugs disappoint for COPD. PMID- 15352284 TI - Managing chronic pain. PMID- 15352285 TI - The next step in vision correction. PMID- 15352286 TI - Rehashing red meat. PMID- 15352287 TI - Avoid angioplasty by lowering cholesterol? PMID- 15352288 TI - Prolapse is less frequent, less dangerous, than previously thought. PMID- 15352290 TI - Alcohol and diabetes. PMID- 15352289 TI - Lose the fat, keep the bone. PMID- 15352291 TI - Surviving a bone marrow transplant. PMID- 15352292 TI - Routine screening for lung cancer? PMID- 15352293 TI - Fighting foot and toenail fungus. PMID- 15352294 TI - A new geranylgeranyl diphosphate synthase gene from Ginkgo biloba, which intermediates the biosynthesis of the key precursor for ginkgolides. AB - Geranylgeranyl diphosphate synthase (GGPPS, EC: 2.5.1.29) catalyzes the biosynthesis of geranylgeranyl diphosphate (GGPP), which is a key precursor for ginkgolide biosynthesis. Here we reported for the first time the cloning of a new full-length cDNA encoding GGPPS from the living fossil plant Ginkgo biloba. The full-length cDNA encoding G. biloba GGPPS (designated as GbGGPPS) was 1657bp long and contained a 1176bp open reading frame encoding a 391 amino acid protein. Comparative analysis showed that GbGGPPS possessed a 79 amino acid transit peptide at its N-terminal, which directed GbGGPPS to target to the plastids. Bioinformatic analysis revealed that GbGGPPS was a member of polyprenyltransferases with two highly conserved aspartate-rich motifs like other plant GGPPSs. Phylogenetic tree analysis indicated that plant GGPPSs could be classified into two groups, angiosperm and gymnosperm GGPPSs, while GbGGPPS had closer relationship with gymnosperm plant GGPPSs. PMID- 15352295 TI - World Health Organization says failure to deliver AIDS medicines is a global health emergency. PMID- 15352296 TI - Diabetes cases could double in developing countries in next 30 years. PMID- 15352297 TI - Global AIDS treatment emergency. PMID- 15352298 TI - WHO/FAO release independent expert report on diet and chronic disease. PMID- 15352299 TI - US $30 million dollars research effort to develop new tests for deadly infectious diseases. PMID- 15352300 TI - WHO launches annual Move for Health day as global initiative to promote benefits of physical activity. PMID- 15352301 TI - PROTEKT US: results from the second year of this longitudinal surveillance study of antimicrobial resistance among respiratory tract pathogens isolated from paediatric and adult patients in the USA. Introduction. PMID- 15352303 TI - Accuracy of a 12-lead electrocardiogram in screening patients with suspected heart failure for open access echocardiography: a systematic review and meta analysis. AB - AIM: The aim of this study was to examine the usefulness of 12-lead electrocardiogram in screening patients with suspected heart failure for open access echocardiography. We carried out a systematic review and meta-analysis of published observational studies. Published studies were identified in the Medline and Embase databases, this was supplemented by key texts, personal reference collections and examining the cited references in the above sources. English language articles in which patients were referred from primary care with a presumptive diagnosis of heart failure to an open access echocardiography service were reviewed. We included studies that reported both electrocardiography findings and echocardiography as a reference standard. Primary outcome measures were sensitivity and specificity, meta-analysed simultaneously to produce a Summary Receiver Operating Characteristic (SROC) curve. RESULTS: Four trials were identified which satisfied the inclusion criteria, including 1419 people with a potential diagnosis of heart failure referred for open access echocardiography. Estimates of sensitivity and specificity varied greatly in individual studies. The area under the SROC curve was 0.84 (95% Cl: 0.33 to 1.00). CONCLUSION: A 12 lead electrocardiogram is an inadequate screening tool for identifying patients with suspected heart failure who require echocardiography. Further research is required to determine the usefulness of other screening strategies including B type natriuretic peptide in screening patients with suspected heart failure who need referral for open access echocardiography. PMID- 15352304 TI - Pathophysiology, evaluation and management of valvular heart disease. Developed from the symposium "Valves in the Heart of the Big Apple", May 2-3, 2002 in New York, New York, USA. PMID- 15352305 TI - Proceedings from the 8th Interdisciplinary Czech-Slovak Toxicological Conference. Praha, Czech Republic, September 3-5, 2003. PMID- 15352306 TI - Festschrift for R G O'Regan - Sensing and adaptation to alterations in respiratory gases: oxygen and carbon dioxide. PMID- 15352307 TI - NAD+ surfaces again. PMID- 15352309 TI - Abstracts of the 58th Annual Meeting of the American Academy for Cerebral Palsy and Developmental Medicine. Los Angeles, California, USA, 2004. PMID- 15352308 TI - SSIEM 41st Annual Symposium. Amsterdam, The Netherlands, 31 August-3 September 2004. Abstracts. PMID- 15352310 TI - Abstracts of the 5th World Congress of Veterinary Dermatology. 25-28 August 2004, Vienna, Austria. PMID- 15352312 TI - Abstracts of the 57th Annual Meeting of the American Association of Blood Banks. Baltimore, Maryland, USA, October 23-26, 2004. PMID- 15352311 TI - Missouri State Medical Association 2004-2005 membership directory. PMID- 15352313 TI - American Society of Animal Science 2004 sectional meeting abstracts. PMID- 15352314 TI - National survey finds thousands of children with mental illness "warehoused" in juvenile detention centers. PMID- 15352315 TI - Genotoxicological studies in aquatic organisms: an overview. AB - Substantial progress has been made in the lat two decades to evaluate the impact of physical and chemical genotoxins in aquatic organisms. This overview (a) summarises the major high lights in this stimulating area of research, (b) compares the developments in this field with the developments in mammalian genotoxicological studies, where appropriate, (c) introduces 18 different articles presented in this special issue of Mutation Research in the backdrop of main advances and , (d) hypothesises on future directions of research in this exciting field. PMID- 15352316 TI - You are what you eat. PMID- 15352317 TI - Preventing unintended pregnancy in the U.S. AB - In 2000, 34 million women--half of all U.S. women of reproductive age--were in need of contraceptive services and supplies to help prevent unintended pregnancy, and half of those were in need of public support for such care. In the absence of a national health insurance program, the United States relies on a patchwork system of public insurance and subsidized clinics to provide care to those in need. Holes in this patchwork, however, are becoming increasingly evident, threatening the system's ability to provide needed services and heightening political interest in more comprehensive solutions. PMID- 15352318 TI - The benefits of investing in sexual and reproductive health. AB - The full value of investing in sexual and reproductive health services has been underestimated, as its wide range of benefits has been largely unrecognized. The direct medical benefits of preventing unintended pregnancies, improving maternal health and preventing, diagnosing and treating sexually transmitted infections including HIV/AIDS are well-known; however, the economic and social benefits are no less real, even if they are more difficult to measure. The global community cannot afford not to fully fund these services to achieve global development goals. PMID- 15352320 TI - Trichonosema algonquinensis n. sp. (Phylum microsporidia) in Pectinatella magnifica (Bryozoa: phylactolaemata) from Algonquin Park, Ontario, Canada. AB - A new species of microsporidian, Trichonosema algonquinensis, is described from a freshwater bryozoan, Pectinatella magnifica from Ontario, Canada. The parasite develops in epithelial cells and appears as white, spherical masses throughout the tissues. Trichonosema algonquinensis is diplokaryotic, diploblastic and undergoes development in direct contact with the cytoplasm of the host cell. Mature spores are ovoid, tapered at one end, and measure 8.5 +/- 0.3 x 4.4 +/- 0.1 microm. The polar filament is wound in 20 to 23 helical coils. Although the parasite resembles T. pectinatellae described from the same host in Michigan and Ohio, it differs in the length of the spore and number of coils of the polar filament. Analysis of 16S rDNA by maximum likelihood, parsimony and Baysian inference, complements the morphological data in supporting the placement of T. algonquinensis as a sister species of T. pectinatellae. PMID- 15352319 TI - Current progress in the fatty acid metabolism in Cryptosporidium parvum. AB - Cryptosporidium parvum is one of the apicomplexans that can cause severe diarrhea in humans and animals. The slow development of anti-cryptosporidiosis chemotherapy is primarily due to the poor understanding on the basic metabolic pathways in this parasite. Many well-defined or promising drug targets found in other apicomplexans are either absent or highly divergent in C. parvum. The recently discovered apicoplast and its associated Type II fatty acid synthetic enzymes in Plasmodium, Toxoplasma, and Eimeria apicomplexans are absent in C. parvum, suggesting this parasite is unable to synthesize fatty acids de novo. However, C. parvum possesses a giant Type I fatty acid synthase (CpFAS1) that makes very long chain fatty acids using mediate or long chain fatty acids as precursors. Cryptosporidium also contains a Type I polyketide synthase (CpPKS1) that is probably involved in the production of unknown polyketide(s) from a fatty acid precursor. In addition to CpFAS1 and CpPKS1, a number of other enzymes involved in fatty acid metabolism have also been identified. These include a long chain fatty acyl elongase (LCE), a cytosolic acetyl-CoA carboxylase (ACCase), three acyl-CoA synthases (ACS), and an unusual "long-type" acyl-CoA binding protein (ACBP), which allows us to hypothetically reconstruct the highly streamlined fatty acid metabolism in this parasite. However, C. parvum lacks enzymes for the oxidation of fatty acids, indicating that fatty acids are not an energy source for this parasite. Since fatty acids are essential components of all biomembranes, molecular and functional studies on these critical enzymes would not only deepen our understanding on the basic metabolism in the parasites, but also point new directions for the drug discovery against C. parvum and other apicomplexan-based diseases. PMID- 15352321 TI - The microtubule analog protein, FtsZ, in the endosymbiont of trypanosomatid protozoa. AB - Blastocrithidia culicis and Crithidia deanei are trypanosomatids that harbor an endosymbiotic bacterium in their cytoplasm. In prokaryotes, numerous proteins are essential for cell division, such as FtsZ, which is encoded by filament-forming temperature-sensitive (fts) genes. FtsZ is the prokaryotic homolog of eukaryotic tubulin and is present in bacteria and archaea, and has also been identified in mitochondria and chloroplasts. FtsZ plays a key role in the initiation of cytokinesis. It self-assembles into the Z ring, which establishes the division plane during septation. In this study, immunoblotting analysis using a FtsZ polyclonal antibody, revealed a 40-kDa band characteristic of FtsZ in endosymbiont fractions and in whole trypanosomatid homogenates, but not in whole cell extracts of aposymbiotic strains. Confocal microscopy and ultrastructural analysis revealed a specific and dispersed labeling over the endosymbiont. Bars and ring-like structures, which are suggestive of the presence of Z-rings, were never observed, even during the division of the symbiont. This peculiar distribution of FtsZ may represent an arrangement of cytoskeleton protein intermediate between prokaryotic and eukaryotic cells. The endosymbiont ftsz gene was completely sequenced after amplification of DNA from symbiont-bearing trypanosomatids or from pure endosymbiont fractions, using PCR and specific primers. The sequences obtained from the endosymbionts from C. deanei and B. culicis were very similar, and were most closely related to bacteria from the genus Pseudomonas. PMID- 15352322 TI - Ribosomal RNA phylogeny of bodonid and diplonemid flagellates and the evolution of euglenozoa. AB - Euglenozoa is a major phylum of excavate protozoa (comprising euglenoids, kinetoplastids, and diplonemids) with highly unusual nuclear, mitochondrial, and chloroplast genomes. To improve understanding of euglenozoan evolution, we sequenced nuclear small-subunit rRNA genes from 34 bodonids (Bodo, Neobodo, Parabodo, Dimastigella-like, Rhynchobodo, Rhynchomonas, and unidentified strains), nine diplonemids (Diplonema, Rhynchopus), and a euglenoid (Entosiphon). Phylogenetic analysis reveals that diplonemids and bodonids are more diverse than previously recognised, but does not clearly establish the branching order of kinetoplastids, euglenoids, and diplonemids. Rhynchopus is holophyletic; parasitic species arose from within free-living species. Kinetoplastea (bodonids and trypanosomatids) are robustly holophyletic and comprise a major clade including all trypanosomatids and most bodonids ('core bodonids') and a very divergent minor one including Ichthyobodo. The root of the major kinetoplastid clade is probably between trypanosomatids and core bodonids. Core bodonids have three distinct subclades. Clade 1 has two distinct Rhynchobodo-like lineages; a lineage comprising Dimastigella and Rhynchomonas; and another including Cruzella and Neobodo. Clade 2 comprises Cryptobia/ Trypanoplasma, Procryptobia, and Parabodo. Clade 3 is an extensive Bodo saltans species complex. Neobodo designis is a vast genetically divergent species complex with mutually exclusive marine and freshwater subclades. Our analysis supports three phagotrophic euglenoid orders: Petalomonadida (holophyletic), Ploeotiida (probably holophyletic), Peranemida (paraphyletic). PMID- 15352323 TI - Co-infection of the macronucleus of Paramecium caudatum by free-living bacteria together with the infectious Holospora obtusa. AB - Infection experiments were performed incubating Paramecium caudatum with non infectious free-living bacteria or weakly infectious intracellular bacteria together with the infectious Holospora obtusa. Two of four non-infectious free living bacteria (Enterobacter aerogenes and Klebsiella pneumoniae) were found to get into the nuclei when added to Paramecium together with H. obtusa. The endonuclear bacterium Nonospora macronucleata that is weakly infectious by itself increases its infectivity when presented together with the infectious holosporas. The results provide evidence that H. obtusa may facilitate entry of other, non infectious bacteria into the nuclei of Paramecium. PMID- 15352324 TI - Inhibition of Vorticella microstoma stalk formation by wheat germ agglutinin. AB - Fluorescently labeled conjugates of wheat germ agglutinin and concanavalin A stained the contractile stalk but not the cell body of Vorticella microstoma trophonts. Binding of the fluorescent conjugants did not noticeably alter the activity of the trophonts. However, unconjugated wheat germ agglutinin prevented free swimming telotrochs from adhering to a glass surface and deploying a contractile stalk during differentiation into trophonts. These observations indicated that the stalk, the material that binds the stalk to surfaces, and the precursors for these components have saccharide residues in common. PMID- 15352325 TI - Identification and assignment of function to the genes of Aspergillus fumigatus expressed at 37 degrees C. AB - Aspergillus fumigatus, a fungal pathogen, causes a spectrum of allergic and invasive disorders. In order to rapidly identify genes of this fungus relevant for pathogenesis and as potential antifungal drug targets, 125 expressed sequence tags (ESTs) were generated from 200 phage clones of a non-normalized cDNA library. Out of a novel 68 ESTs, 45 were assigned putative functions based on the sequence similarity. The identities of some of these genes suggest that they may be involved in pathogenesis or autoimmune reactions. Additional genes were identified that are possible targets for the development of antifungal drugs or that may be of use in diagnosing fungal infections. PMID- 15352326 TI - A molecular biological approach to the phylogenetic position of the genus Hyperamoeba. AB - In 1923 Alexeieff described a new amoebic species within a new genus and named it Hyperamoeba flagellata. This amoeba exhibits three life cycle stages, an amoeboid trophozoite, a flagellated stage, and a cyst-like the heteroloboseans, the mastigamoebae, and several slime moulds. Since then more strains have been isolated and relationships to the protostelids and cercomonads and to the myxogastrid plasmodial slime moulds have been suggested. However, up to now the classification and phylogenetic position of the hyperamoebae has remained unclear. The aim of our study was to make an approach to the phylogeny of the genus Hyperamoeba with combined morphological and molecular biological data. Since 1988 we have isolated and collected Hyperamoeba-like strains from different aquatic and terrestrial sources. The 18S rDNA-sequences of 8 new Hyperamoeba strains isolated from various habitats were analysed and a cluster analysis was performed including all other available hyperamoebae. Altogether, the results of our study corroborate the relatedness of Hyperamoeba to various slime moulds. However, the hyperamoebae do not seem to be a monophyletic group, clearly putting the validity of the genus Hyperamoeba into question. PMID- 15352327 TI - Molecular phylogeny of phyllopharyngean ciliates and their group I introns. AB - We analyzed small subunit ribosomal DNA (ssu-rDNA) sequences to evaluate both the monophyly of the ciliate class Phyllopharyngea de Puytorac et al. (1974), and relationships among subclasses. Classifications based on morphology and ultrastructure divide the Phyllopharyngea into four subclasses, the Phyllopharyngia, Chonotrichia, Rhynchodia, and Suctoria. Our analyses of ssu-rDNA genealogies derived from sequence data collected from diverse members representing three of the four subclasses of Phyllopharyngea (Suctoria: Ephelota spp., Prodiscophyra collini, Acineta sp.; Phyllopharyngia: Chlamydodon exocellatus, Chlamydodon triquetrus, Dysteria sp.; and Chonotrichia: Isochona sp.) provide strong support for the monophyly of the Phyllopharyngea, and show that the Chonotrichia emerge from within the Phyllopharyngia. Based on this initial sampling, suctorian budding types are monophyletic, and exogenous budding appears to be basal to evaginative and endogenous budding. Further, we report the discovery of a group I intron at position 891 in the Suctoria Acineta sp. and Tokophrya lemnarum, and a second group I intron at position 1506 in T. lemnarum. These introns represent only the second examples of group I introns in a ciliate ribosomal gene, since the discovery of ribozymes in the LSU rRNA gene of Tetrahymena thermophila. Phylogenetic analyses of Group I introns suggest a complex evolutionary history involving either multiple loses or gains of introns within endogenously budding Suctoria. PMID- 15352328 TI - Strombidinopsis jeokjo n. sp. (ciliophora: choreotrichida) from the coastal waters off western Korea: morphology and small subunit ribosomal DNA gene sequence. AB - The planktonic ciliate Strombidinopsis jeokjo n. sp. is described from Quantitative Protargol-Stained (QPS) preparations, and the sequence of the small subunit rDNA (SSU rDNA) from cultured cells is reported. This species is ovoid and bluntly tapered towards the posterior. The ranges (and mean +/- standard deviation, n = 31) of cell length, cell width, and oral diameter of the QPS stained specimens were 100-190 microm (149 +/- 25), 60-105 microm (79 +/- 13), and 55-80 microm (64 +/- 5), respectively. Fifteen to seventeen external oral polykinetids had oral membranelle cilia 20-35 microm long. Twenty-six to twenty eight somatic kineties were equally spaced around the cell body and extended from the oral to the posterior regions with 23-44 dikinetids per kinety. Both kinetosomes of each kinetid bore cilia 3-7 microm long. Strombidinopsis jeokjo had two ovoid macronuclei of 25-38 microm x 12-15 microm. When properly aligned, the sequence of the SSU rDNA of S. jeokjo (GenBank Accession No. AJ628250) was approximately 2% different from that of an unidentified Strombidinopsis species (GenBank Accession No. AF399132-AF399135), the closest species in the SSU rDNA sequence. PMID- 15352329 TI - Pyruvate formate lyase (PFL) and PFL activating enzyme in the chytrid fungus Neocallimastix frontalis: a free-radical enzyme system conserved across divergent eukaryotic lineages. AB - Fermentative formate production involves the activity of pyruvate formate lyase, an oxygen-sensitive enzyme that employs a glycyl radical in its reaction mechanism. While common among anaerobic prokaryotes, this enzyme has so far been found in only two distantly related eukaryotic lineages, anaerobic chytridiomycetes and chlorophytes. Sequence comparisons of homologues from the chytridiomycetes Piromyces and Neocallimastix, the chlorophyte Chlamydomonas, and numerous prokaryotes suggest a single, eubacterial origin of eukaryotic pyruvate formate lyases. Pyruvate formate lyase activating enzyme introduces the glycyl radical into the pyruvate formate lyase protein chain. We discovered this enzyme, which had not previously been reported from eukaryotes, in the same two eukaryotic lineages and show that it shares a similar evolutionary history to pyruvate formate lyase. Sequences with high homology to pyruvate formate lyase activating enzyme were identified in the genomes of the anaerobic protozoan parasites Trichomonas vaginalis, Entamoeba histolytica, and Giardia intestinalis. While the occurrence of pyruvate formate lyase activating enzyme together with pyruvate formate lyase in fungi and chlorophytes was to be expected, the target protein of a glycyl radical enzyme-activating enzyme in these protozoa remains to be identified. PMID- 15352330 TI - Predicted secondary structure of the foraminiferal SSU 3' major domain reveals a molecular synapomorphy for granuloreticulosean protists. AB - The small subunit ribosomal RNA genes of foraminiferal protists are the largest and most divergent of any eukaryote. We demonstrate that this foraminiferal sequence alteration represents a substantial modification to the small subunit ribosomal RNA structure, including a large (up to 350 nt) novel helix in a very well-conserved portion of the head domain. This modification dates from the beginning of the foraminiferal radiation and, within modern orders, is partially conserved at the sequence level, suggesting that it is a functional part of the ribosome. The pattern of conservation makes it particularly useful for determining lower-taxon relationships in morphologically ambiguous allogromiid foraminifera. PMID- 15352332 TI - Vertical micro-distribution and response to nitrogen deposition of testate amoebae in Sphagnum. AB - Previous studies have shown the existence of a vertical micro-distribution of testate amoebae in the first centimeters of Sphagnum and their response to nutrient enrichment. In order to test the response of testate amoebae to depth and N addition in dry moss carpets recolonizing cutover peatlands, we sampled Sphagnum that had received 0, 1, 3, or 10g N m(-2) yr(-1) for three years. The mosses were cut into three segments: 0-1cm, 1-3cm and 3-5cm and analyzed for testate amoebae. The overall diversity (22 taxa) was high considering the dryness of the site, but the species richness of individual samples was low (mean 6.6). The presence of several species characteristic of wetter conditions suggests that they have a broader tolerance than usually believed and/or have a high colonization potential. Species richness increased with depth. Assulina muscorum was most abundant in the top segment, while Phryganella acropodia, Heleopera rosea and Nebela militaris were most abundant in the deepest segment. Neither the metabolism type nor the shell characteristics significantly explained the vertical distribution of species. There was no overall response of testate amoebae to N, although one species, Bullinularia indica, was significantly more abundant in the fertilized than in the control plots. PMID- 15352331 TI - Differential stimulation of microglial pro-inflammatory cytokines by Acanthamoeba culbertsoni versus Acanthamoeba castellanii. AB - Acanthamoeba spp. are opportunistic pathogens that cause granulomatous amebic encephalitis. We compared the highly pathogenic species A. culbertsoni to the relatively less pathogenic species A. castellanii for its capacity to elicit from neonatal rat microglia the gene expression of pro-inflammatory cytokines. Acanthamoeba culbertsoni elicited a robust cytokine gene response by neonatal rat microglia in vitro as compared to A. castellanii. The preponderant cytokine elicited at the mRNA and protein levels was interleukin-1beta. In addition, transmission electron microscopy revealed that microglial cells were capable of phagocytozing A. castellanii. In contrast, A. culbertsoni destroyed microglia. Collectively, these results suggest that a combined action of pro-inflammatory cytokines and destruction of host cells by amebae contribute to the pathology caused by the more pathogenic species. PMID- 15352333 TI - Thwarting behavioral health violence through facility design. PMID- 15352334 TI - The Rashomon effect: organization ethics in health care. AB - The Academy-Award winning 1950 Japanese movie Rashomon depicts an incident involving an outlaw, a rape or seduction of a woman, and a murder or suicide of her husband. A passer-by, who is also the narrator, explains how the story is told to officials from four different perspectives: that of the outlaw, the woman, the husband and himself. The four narratives agree that the outlaw, wandering through the forest, came upon the woman on a horse being led by her husband; the outlaw tied up the husband, sex took place between the woman and the outlaw in front of the bound husband, and the husband was found dead. The narratives do not agree on how these events occurred or who killed the husband. The outlaw contends that consensual sex occurred between him and the wife, and he claims to have killed the husband. The wife depicts the sex as rape and claims that because of her disgrace she killed her husband. The husband, through a medium, says that the sexual act began as rape and ended in consent, and that in shame, after being untied by the outlaw he killed himself. The passer-by's story agrees with the husband's account of the sex and the bandit's account of the murder. Because the passer-by is also the narrator of the film, we tend to believe his version. But what actually took place is never resolved. PMID- 15352335 TI - Criteria for determining the appropriate method for an ethics consultation. PMID- 15352336 TI - Seeking common ground in a world of ethical pluralism: a review essay of Moral Acquaintances: methodology in bioethics by Kevin Wm. Wildes, S.J. PMID- 15352337 TI - Normal variants of competence to consent to treatment. PMID- 15352338 TI - Canadian Medical Association's ethics activities. PMID- 15352339 TI - Validating a patient satisfaction survey translated into Spanish. AB - A measure of patient satisfaction with the inpatient care experience, which was originally developed in English, was validated in Spanish, and differences in satisfaction between English-language and Spanish-language respondents were assessed. The Spanish translation of the survey demonstrated the same underlying factor structure of the English version of the survey, was reliable at both the subscale and overall level, and accounted for 81% of the variance in Spanish language respondents' reported likelihood to recommend the hospital to others. Spanish-language respondents showed higher mean satisfaction levels with regard to most aspects of their care than English-language respondents, except when rating staff courtesy. PMID- 15352340 TI - Improving the quality of diabetes care in primary care practice. AB - The quality of care delivered to patients with diabetes has an impact on long term outcomes. The purpose of this quality improvement project was to examine the effect of a Diabetes Disease Management Program (DDMP) on compliance with recommended process measures of care in primary care practice settings. Certified diabetes nurse educators visited five participating primary care practices biweekly for 1 year providing education to physicians and office staff on standards for diabetes management and to patients regarding self-management. Among 208 participants, 85.5% had >2 HbA1cs, 93.6% had lipid profiles, and 100% had foot examinations. Improvements in HbA1c (9.0% to 7.7%) were observed. Implementation of DDMP effectively promotes compliance with care standards in primary care practices. PMID- 15352341 TI - Carolyn M. Clancy on the Agency for Healthcare Research and Quality. PMID- 15352342 TI - Can't miss: conquer any number task by making important statistics simple. Part 4. Confidence intervals with t distributions, standard error, and confidence intervals for proportions. AB - Healthcare quality professionals need to understand and use inferential statistics to interpret sample data from their organizations. Since in quality improvement and healthcare research studies, all the data from a population often are not available, investigators take samples and make inferences about that population using inferential statistics. This series of six articles will give readers an understanding of the concepts of inferential statistics, as well as the specific tools for calculating confidence intervals and tests of statistical significance for samples of data. The statistical principles are equally applicable to quality improvement and healthcare research studies. This article, Part 4, starts with a review of the information contained in Parts 1, 2, and 3, which appeared in the July/August 2003 issue of the Journal for Healthcare Quality. This article describes t distributions and how these are used to calculate confidence intervals for estimating a population mean based on a sample mean of a continuous variable. Part 4 concludes with a discussion of standard error, margin of error, and confidence intervals for estimating a population proportion based on a sample proportion from a binomial variable. PMID- 15352343 TI - Can't miss: conquer any number task by making important statistics simple. Part 5. Comparing two confidence intervals, standard error of the difference between two means and between two proportions. AB - Healthcare quality professionals need to understand and use inferential statistics to interpret sample data from their organizations. Since in quality improvement and healthcare research studies all the data from a population often are not available, investigators take samples and make inferences about that population using inferential statistics. This series of six articles will give readers an understanding of the concepts of inferential statistics as well as the specific tools for calculating confidence intervals and tests of statistical significance for samples of data. This article, Part 5, demonstrates the comparison of two confidence intervals as a method for estimating the difference between two population means. The concept of the standard error of the difference between two sample means is presented along with the confidence interval for estimating the difference between two population means. The article concludes with the standard error and confidence interval for estimating the difference between two population proportions from a binomial variable. PMID- 15352344 TI - Can't miss: conquer any number task by making important statistics simple. Part 6. Tests of statistical significance (z test statistic, rejecting the null hypothesis, p value), t test, z test for proportions, statistical significance versus meaningful difference. AB - Healthcare quality professionals need to understand and use inferential statistics to interpret sample data from their organizations. Since in quality improvement and healthcare research studies all the data from a population often are not available, investigators take samples and make inferences about that population using inferential statistics. This series of six articles will give readers an understanding of the concepts of inferential statistics as well as the specific tools for calculating confidence intervals and tests of statistical significance for samples of data. This article, Part 6, merges the four concepts of the (1) standard error of the difference between sample means, (2) the z test statistic, (3) rejecting the null hypothesis, and (4) the p value to provide a comprehensive view of tests of statistical significance. This is followed by a description of t tests, statistical tests for comparing two sample proportions, and Type I and Type II errors. The series of articles concludes with a description of statistical significance versus meaningful difference. PMID- 15352345 TI - Superior signal-to-noise ratio of a new AA1 sequence for random-modulation continuous-wave lidar. AB - In an earlier work [Proc. SPIE 4484, 216 (2001)] we proposed a new AA1 modulation sequence for random-modulation continuous-wave lidar. It possesses significantly better signal properties than other pseudorandom codes (the M, A1, and A2 sequences). We derive and compare the signal-to-noise ratio (SNR) of the new AA1 sequence with those of previous modulation sequences. Using a figure of merit proposed for pseudorandom sequences in additive (and generally colored) noise, we show that the SNR of the AA1 sequence in 1/f noise can be as much as 50 times better than that of the commonly used M sequence. This improved SNR should permit as much as a 7:1 increase of the maximum lidar sensing range in baseband modulation direct-detection infrared lidar with no significant changes to the transmitter and receiver. PMID- 15352346 TI - Comment on "Complete electromagnetic coherence in the space-frequency domain". AB - It is pointed out that a recently introduced definition of the degree of coherence of a random electromagnetic field is not a true quantitative measure of coherence. PMID- 15352347 TI - Polarization singularity democracy: WYSIWYG. AB - The canonical point singularity of elliptically polarized light is a C point, an isolated point of circular polarization surrounded by a field of polarization ellipses. The defining singular property of a C point is that the surrounding ellipses rotate about the point. It is shown that this rotation is seen only for a particular line of sight (LOS) and, conversely, that there exists a unique LOS for every ellipse along which the ellipse is seen as a singularity. It is also shown that changes in LOS can turn singularities into stationary points and vice versa. The democratic behavior of polarization singularities and stationary points is a consequence of the fundamental "what you see is what you get" property of ellipse fields. Simple experiments are proposed for observing this unusual property of elliptically polarized light. PMID- 15352348 TI - Engineering space-variant inhomogeneous media for polarization control. AB - Novel devices for converting a linear polarization state to radial or azimuthal polarization states are realized by use of space-variant inhomogeneous media on a subwavelength scale. The two designs presented use form birefringence to locally transform the polarization state. The devices are fabricated in a GaAs substrate for operation in the far-infrared wavelength range. The experimental characterization is in good agreement with the designs, demonstrating high conversion efficiency. PMID- 15352349 TI - Consequences of the Kramers-Kronig relations for light diffraction on thick gratings. AB - Diffraction of monochromatic light on a grating leads to the attenuation of the transmitted beam of diffraction order zero. In the case of a thick grating the diffraction efficiency, and hence the effective attenuation coefficient, is a fast-varying function of the Bragg mismatch angle. According to Kramers-Kronig theory, the transmitted beam encounters a phase shift that also depends on the mismatch angle. This phase shift is measured for holographic gratings in a photoaddressable block copolymer and compared with analytical calculations. PMID- 15352350 TI - Elegant Ince-Gaussian beams. AB - The existence of elegant Ince-Gaussian beams that constitute a third complete family of exact and biorthogonal elegant solutions of the paraxial wave equation is demonstrated. Their transverse structure is described by Ince polynomials with a complex argument. Elegant Ince-Gaussian beams constitute exact and continuous transition modes between elegant Laguerre-Gaussian and elegant Hermite-Gaussian beams. The expansion formulas among the three elegant families are derived. PMID- 15352351 TI - Fast profile measurement of micrometer-sized tapered fibers with better than 50 nm accuracy. AB - The forward scattering of light illuminating a transparent dielectric cylinder, such as a tapered fiber, from the side can be understood as interference of the diffracted, reflected, and transmitted light. Additionally, light can be resonantly coupled into the fiber if a multiple of the wavelength matches the circumference. Using a suitable laser setup with a novel evaluation algorithm allows us to quickly extract the fiber radius from the complex diffraction pattern, obtaining an accuracy of better than 50 nm. We demonstrate experimentally our method, which is noncontact and allows one to simultaneously measure the profile of a several-centimeter-long fiber waist with a diameter near the diffraction limit. PMID- 15352352 TI - Generation of pure two-beam interference grating structures in an optical fiber with a femtosecond infrared source and a phase mask. AB - Fiber Bragg gratings were fabricated in all-silica core fiber by focusing 125-fs 800-nm pulses with an 80-mm lens through a phase mask with 4.28-microm pitch onto a fiber sample. When the phase-mask-fiber separation was 5 mm the observed structure was clearly the result of two-beam interference between the +/- 1 orders. The elimination of the remaining 9 orders is a consequence of the walk off experienced by the mask orders and the short duration of the femtosecond pulse. This effect is unique to the fabrication of Bragg gratings with femtosecond sources and would not be observed with a longer pulse duration or incoherent UV sources. PMID- 15352353 TI - Frequency stabilization of a 1319-nm Nd:YAG laser by saturation spectroscopy of molecular iodine. AB - Hyperfine transitions of molecular iodine were observed by use of the frequency doubled output of a 1319-nm Nd:YAG laser with saturation spectroscopy. The laser frequency was stabilized to the observed hyperfine transition and reached a stability of 6 x 10(-12) for a 1.5-s averaging time, improving toward the 1 x 10( 12) level after 100 s. The iodine-stabilized 1319-nm Nd:YAG laser is an excellent candidate for an optical frequency standard for telecommunication applications. PMID- 15352354 TI - Signal-processing algorithm for white-light optical fiber extrinsic Fabry-Perot interferometric sensors. AB - We present a novel signal-processing algorithm for single-mode optical fiber extrinsic Fabry-Perot interferometric sensors that can achieve both high resolution, absolute measurement of the cavity length and a large dynamic measurement range simultaneously. The algorithm is based on an accurate model of the characteristics of a fiber-optic sensor that takes into account the phase shift that is due to the coupling of light reflected at the second surface to the lead-in fiber end. PMID- 15352355 TI - Applicability of classical optical fiber theories to holey fibers. AB - We discuss the applicability of well-established classical optical fiber theories to holey fibers. By appropriately defining the V parameter, we can easily estimate the fundamental properties of holey fibers, such as effective index, group-velocity dispersion, mode field diameter, beam divergence, and splice loss, through simple empirical expressions without the need for heavy numerical computations. We confirm the validity of the V parameter defined here by comparing the calculated results with the earlier experimental and numerical results. PMID- 15352356 TI - Numerical analysis of an annular-aperture solid immersion lens. AB - A physical model of an annular-aperture solid immersion lens (SIL) is proposed, and its attractive features are presented numerically with the finite-difference time-domain method. Placing an appropriate annular aperture in front of the SIL shows that the focal depth can evidently be improved, combining the virtues of the annular-aperture technique and the SIL technique. With this proposed method the rigorous distance control condition in related devices can be relaxed, preventing scratches or collisions between the optical head and the recording medium. Potentially, this technique could have great prospects for applications in optical data recording, lithography, and other applications that depend on immersion media to meet the resolution criteria across the image field. PMID- 15352357 TI - High-bandwidth transmission of an efficient photonic-crystal mode converter. AB - We have investigated both theoretically and experimentally the spectral behavior and the transmission and reflection performance of a photonic-crystal (PhC) mode converter upon an InP substrate. This taper exhibits 70% transmission efficiency on an 80-nm bandwidth when it couples a ridge access guide to a strongly confined single-missing-row PhC guide. Such a taper design included in a PhC bend contributes a large benefit to the overall transmission budget of the PhC-based link. PMID- 15352358 TI - Total internal reflection-evanescent coupler for fiber-to-waveguide integration of planar optoelectric devices. AB - We present a method for parallel coupling from a single-mode fiber, or fiber ribbon, into a silicon-on-insulator waveguide for integration with silicon optoelectronic circuits. The coupler incorporates the advantages of the vertically tapered waveguides and prism couplers, yet offers the flexibility of planar integration. The coupler can be fabricated by use of either wafer polishing technology or gray-scale photolithography. When optimal coupling is achieved in our experimental setup, the coupler can be packaged by epoxy bonding to form a fiber-waveguide parallel coupler or connector. Two-dimensional electromagnetic calculation predicts a coupling efficiency of 77% (- 1.14-dB insertion loss) for a silicon-to-silicon coupler with a uniform tunnel layer. The coupling efficiency is experimentally achieved to be 46% (-3.4-dB insertion loss), excluding the loss in silicon and the reflections from the input surface and the output facet. PMID- 15352359 TI - Broadband generation of terahertz radiation in a waveguide. AB - We report a novel geometry that allows for the phase-matched generation of broadband terahertz radiation in a polymer-based parallel-plate metal waveguide by means of optical rectification. Both the optical pump beam and the generated terahertz radiation propagate in the fundamental mode of the waveguide. This allows for noncritical phase matching over a broad range of terahertz frequencies. We demonstrate guided-wave interaction lengths of up to 3 mm. PMID- 15352360 TI - Micromachined array tip for multifocus fiber-based optical coherence tomography. AB - High-resolution optical coherence tomography demands a large detector bandwidth and a high numerical aperture for real-time imaging, which is difficult to achieve over a large imaging depth. To resolve these conflicting requirements we propose a novel multifocus fiber-based optical coherence tomography system with a micromachined array tip. We demonstrate the fabrication of a prototype four channel tip that maintains a 9-14-microm spot diameter with more than 500 microm of imaging depth. Images of a resolution target and a human tooth were obtained with this tip by use of a four-channel cascaded Michelson fiber-optic interferometer, scanned simultaneously at 8 kHz with geometric power distribution across the four channels. PMID- 15352361 TI - Photoacoustic and ultrasonic coimage with a linear transducer array. AB - A technique is developed to simultaneously acquire ultrasound and photoacoustic (PA) images based on a linear transducer array. The system uses conventional ultrasound for rapid identification of potential targets. After a target is identified, the ultrasound echo and PA signals can be simultaneously obtained with optimized excitation and a signal collection sequence. The corresponding ultrasound impedance and optical absorption images are reconstructed with a phase controlled algorithm. The approach can effectively reduce the artifacts associated with a conventional filter backprojection algorithm used in PA imaging by linear scanning. The technique provides a new approach for practical applications. PMID- 15352362 TI - Rapid-scanning forward-imaging miniature endoscope for real-time optical coherence tomography. AB - We developed a miniature endoscope that is capable of rapid lateral scanning and is suitable for real-time forward-imaging optical coherence tomography (OCT). The endoscope has an outer diameter of 2.4 mm, consisting of a miniature tubular lead zirconate titanate (PZT) actuator, a single-mode fiber-optic cantilever, and a graded-index lens. Rapid lateral scanning at 2.8 kHz is achieved when the fiber optic cantilever is resonated with the PZT actuator. This allows OCT imaging to be performed by fast lateral beam scanning followed by slow depth scanning, which is different from the conventional OCT imaging sequence. Real-time OCT imaging with the endoscope operated in the new image acquisition sequence at 6 frames/s is demonstrated. PMID- 15352363 TI - Diffuse optical measurement of blood flow, blood oxygenation, and metabolism in a human brain during sensorimotor cortex activation. AB - We combine diffuse optical and correlation spectroscopies to simultaneously measure the oxyhemoglobin and deoxyhemoglobin concentration and blood flow in an adult human brain during sensorimotor stimulation. The observations permit calculation of the relative cerebral metabolic rate of oxygen in the human brain, for the first time to our knowledge, by use of all-optical methods. The feasibility for noninvasive optical measurement of blood flow through the skull of an adult brain is thus demonstrated, and the clinical potential of this hybrid, all-optical noninvasive, methodology can now be explored. PMID- 15352364 TI - Defect-mode dependence of two-photon-absorption enhancement in a one-dimensional photonic bandgap structure. AB - A one-dimensional photonic crystal containing a single CdS defect layer of various thicknesses was fabricated. The dependence of the two-photon-absorption (TPA) coefficient on the defect mode was investigated by use of a femtosecond pump-probe method. Experimental results show that the TPA coefficient of the CdS defect layer depends strongly on the defect mode in the photonic bandgap. This is consistent with the predicted dependence of light intensity within the defect layer. PMID- 15352365 TI - Control of multiple filamentation in air. AB - In this Letter we provide what is believed to be the first experimental evidence of suppression of the number of filaments for high-intensity laser pulses propagating in air by beam astigmatism. We also show that the number, pattern, and spatial stability of the filaments can be controlled by varying the angle that a focusing lens makes with the axial direction of propagation. This new methodology can be useful for applications involving atmospheric propagation, such as remote sensing. PMID- 15352366 TI - Vapor-transport equilibrated near-stoichiometric lithium tantalate for frequency conversion applications. AB - Near-stoichiometric lithium tantalate (SLT) crystals were produced from congruent lithium tantalate by a vapor-transport equilibration process. Because of the resultant increase in photoconductivity and reduction in photogalvanism, the crystals showed no observable photorefractive damage at 514.5 nm up to the highest intensity used, 2 MW/cm2. The crystals also exhibited low green-induced infrared absorption, a Curie temperature of 693 degrees C, and a coercive field of 80 V/mm. The SLT samples were periodically poled with an 8-microm-period grating, permitting first-order quasi-phase-matched second-harmonic generation of 532-nm radiation at 43 degrees C. A 17-mm-long sample generated 1.6 W of continuous-wave output power at 532 nm for 50 h. With 150-ns pulses at a 100-kHz repetition rate in the same sample, 5-W average-power, 532-nm radiation was generated for 1000 h. No damage to the crystal and no aging effects were observed during these experiments. PMID- 15352367 TI - Low-threshold spatial solitons in reverse-proton-exchanged periodically poled lithium niobate waveguides. AB - Combining reverse proton exchange and uniform periodic poling in LiNbO3 planar waveguides, we demonstrate low-energy spatial optical solitons by second-harmonic generation at room temperature, with a threshold as low as 23 pJ/microm at 1.5 microm. PMID- 15352368 TI - High-speed horizontal-path atmospheric turbulence correction with a large actuator-number microelectromechanical system spatial light modulator in an interferometric phase-conjugation engine. AB - Results of atmospheric propagation for a high-speed, large-actuator-number adaptive optics system are presented. The system uses a microelectromechanical system- (MEMS-) based spatial light modulator correction device with 1024 actuators. Tests over a 1.35-km path achieved correction speeds in excess of 800 Hz and Strehl ratios close to 0.5. The wave-front sensor was based on a quadrature interferometer that directly measures phase. This technique does not require global wave-front reconstruction, making it relatively insensitive to scintillation and phase residues. The results demonstrate the potential of large actuator-number MEMS-based spatial light modulators to replace conventional deformable mirrors. PMID- 15352369 TI - Demonstration of a simple technique for determining the M/# of a holographic substrate by use of a single exposure. AB - We propose and demonstrate a simple technique for determining the M/# parameter of a holographic recording material. In this method, divergent object and reference beams are used to produce a spatially varying index modulation. One can analyze the resultant diffraction pattern to find M/# by using only a single grating; existing techniques require many gratings. PMID- 15352370 TI - Reconstruction of in-line digital holograms from two intensity measurements. AB - We propose a new method based on in-line digital holography for the reconstruction of a wave front from only two intensity recordings. The simulation result shows that this method works well when the object wave is weak compared with the reference wave. This technique can be employed for real-time imaging. PMID- 15352371 TI - Depth-enhanced integral imaging by use of optical path control. AB - The image depth of integral imaging is enhanced by doubling the number of central depth planes by use of optical path control. To accomplish this, the optical path lengths are changed by controlling whether reflections occur behind the lens array. We propose three schemes that use mirrors, a combination of beam splitters and polarizers, and polarization beam splitters, respectively. In experiments we implement the systems that are completely electronically controllable, are compact, and provide two central depth planes with 50.4-mm separation. PMID- 15352372 TI - Applications of fractional transforms to object reconstruction from in-line holograms. AB - We propose a method for the digital reconstruction of an object whose diffraction pattern has been recorded on a hologram. The fractional Fourier transform is used for the object reconstruction. To determine the position of the object, the fractional order is scanned. The fractional cosine and fractional Hartley transforms are also employed for object reconstruction. These two transforms are real valued and allow the reconstruction to be done with lower computing complexity. Simulations and experimental results are presented. PMID- 15352373 TI - Optical vortex beam shaping by use of highly efficient irregular spiral phase plates for optical micromanipulation. AB - Optical dark traps such as Laguerre-Gaussian beams, modulated optical vortices, and high-order Bessel beams have been used in the micromanipulation of microparticles. Such optical traps are highly versatile, as they are able to trap both high- and low-index microparticles as well as to set them into rotation by use of the orbital angular momentum of light. Holography has been widely used to modulate the shape of an optical vortex for new optical traps. We show that, by designing the shape of a spiral phase plate and using electron-beam lithography for fabrication, one can modulate the amplitude and the phase of an optical vortex with respect to the specific shape of the spiral phase plate as required. Furthermore, to the best of our knowledge this is the first report of transferring orbital angular momentum from a spiral phase plate to an absorptive microparticle in an experiment. Hence, with this technique, optical dark traps can easily be designed and fabricated. PMID- 15352374 TI - Observation of rubidium 5S1/2 --> 7S1/2 two-photon transitions with a diode laser. AB - The 5S1/2 --> 7S1/2 two-photon transition of atomic rubidium, which is 100 times weaker than the 5S-5D transition, is observed with an extended-cavity diode laser and a vapor cell. Signals with a signal-to-noise ratio of 280 are obtained with a laser power of 10 mW, and the observed linewidth is 3 MHz. The laser wavelength is 760 nm and is locked on the transitions to a stability of 2 x 10(-11). For the first time to our knowledge, the isotope shift of this transition is measured to be 130(4) MHz in atomic frequency. PMID- 15352375 TI - Typesetting of terahertz waveforms. AB - We demonstrate programmable generation of temporally shaped terahertz waveforms in LiNbO3 by spatially shaping the beam profile of femtosecond excitation laser pulses with a spatial light modulator. The generated terahertz waveforms have amplitudes that are approximately proportional to the first spatial derivative of the excitation beam profile. PMID- 15352376 TI - Terahertz emission via ultrashort-pulse excitation of magnetic metal films. AB - We observe terahertz emission by optical rectification of an intense 1.5-eV, 50 fs pulse in single-crystal iron thin films grown by molecular beam epitaxy. The azimuthal dependence of the emission indicates the presence of a magnetic nonlinearity and a nonmagnetic surface nonlinearity. PMID- 15352377 TI - Carrier-envelope phase slip of ultrashort dispersion-managed solitons. AB - The carrier-envelope phase slip of an ultrashort pulse circulating in a mode locked Ti:sapphire laser is analyzed. The laser cavity is modeled by a dispersion and nonlinearity-managed nonlinear Schrodinger equation. The combined contributions to the phase slip induced by nonlinear phase and nonlinear dispersion are found to approach zero for strong dispersion maps. The dependence of the slip on third-order dispersion is found as well. The analytical results are verified using numerical simulations. PMID- 15352378 TI - Fourier algorithm for four-wave-mixing signals from optically dense systems with memory. AB - A triple Fourier-transform algorithm for generating and propagating femtosecond four-wave-mixing signals in optically thick samples is demonstrated. The algorithm has a dynamic range that is useful for tests of theory and simulations of experiments with an arbitrary nonlinear response. Although two-pulse echoes decay faster as optical density increases for a Bloch model, we find that systems with memory exhibit the opposite trend. PMID- 15352379 TI - The influence of fasting on liver sulfhydryl groups, glutathione peroxidase and glutathione-S-transferase activities in the rat. AB - Sulfhydryl groups, glutathione peroxidase (GPx) and glutathione-S-transferase (GST) are important elements of the antioxidant defence in the organism. The efficacy of their antioxidant action is influenced by many factors. In this work, the effect of fasting on total, protein-bound and nonprotein sulfhydryl groups and on the activity of liver and serum GPx and GST in rats were determined. Male Wistar rats were divided into two groups: non-fasted and 18-hour fasted. In fasted animals liver content of nonprotein sulfhydryl groups (represented predominantly by reduced glutathione; GSH) was diminished by 22% in comparison to non-fasted group, whereas total and protein-bound -SH groups were unaffected. The activity of liver and serum GPx was unchanged in food deprived rats. In these animals the activity of GST in serum was reduced by 26%. Fasting had no significant effect on the activity of GST in the liver. Our results demonstrate that in rats deprived of food for 18 hours liver and serum GPx and GST are not involved in protection against action of reactive oxygen species formed during fasting. The observed drop in the content of liver nonprotein sulfhydryl groups without concomitant rise in the activity of GPx and GST indicates that this effect may be due to augmented degradation of GSH, its potentiated efflux from hepatocytes and formation of conjugates with intermediates arising as a result of reactive oxygen species action. PMID- 15352380 TI - Construction of chimeric cytosolic fructose-1,6-bisphosphatases by insertion of a chloroplastic redox regulatory cluster. AB - In order to transform cytosolic fructose-1,6-bisphosphatases (FBPase)(EC 3.1.3.11) into potential reductively-modulated chloroplast-type enzymes, we have constructed four chimeric FBPases, which display structural viability as deduced by previous modelling. In the X1-type BV1 and HL1 chimera the N-half of cytosolic sugar beet (Beta vulgaris L.) and human FBPases was fused with the C-half of the pea (Pisum sativum L.) chloroplast enzyme, which carries the cysteine-rich light regulatory sequence. In the X2-type BV2 and HL2 chimera this regulatory fragment was inserted in the corresponding site of the sugar beet cytosolic and human enzymes. Like the plant cytosolic FBPases, the chimeric enzymes show a low rise of activity by dithiothreitol. Both BV1 and BV2, but not HL1 and HL2, display a negligible activation by Trx f, but neither of them by Trx m. Antibodies raised against the pea chloroplast enzyme showed a positive reaction against the four chimeric FBPases and the human enzyme, but not against the sugar beet one. The four chimera display typical kinetics of cytosolic FBPases, with Km values in the 40-140 microM range. We conclude the existence of a structural capacity of cytosolic FBPases for incorporating the redox regulatory cluster of the chloroplast enzyme. However, the ability of these chimeric FBPases for an in vitro redox regulation seems to be scarce, limiting their use from a biotechnology standpoint in in vivo regulation of sugar metabolism. PMID- 15352381 TI - MAO activity in serotonergic endings of rat major cerebral arteries. AB - The present work studies the existence of monoamine oxidase (MAO) activity in serotonergic endings present in rat major cerebral arteries. Enzymatic activity was appraised in vivo by serotonin (5-HT) accumulation or 5-hydroxyindole acetic acid (5-HIAA) disappearance with time after systemic administration of MAO inhibitors. Pargyline (75 mg/Kg, ip) brought about significant 5-HT increase and 5-HIAA decrease in major cerebral arteries 30 and 60 min after its administration. Clorgyline (75 mg/Kg, ip) also induced 5-HT enhancement and 5 HIAA decline in these arteries 30 and 60 min after its injection. However, treatment with deprenyl (75 mg/Kg, ip) only evoked a significant 5-HT increase at 60 min. When either clorgyline (5 mg/Kg, ip) or deprenyl (5 mg/Kg, ip) were administered 5-HT and 5-HIAA levels remained unaffected. Two weeks after performing electrolytical lesion of dorsal raphe nucleus and 60 min after clorgyline (75 mg/Kg, ip) injection 5-HT and 5-HIAA levels appeared significantly reduced in cerebral arteries and striatum when compared to sham-lesioned controls. These results suggest that MAO-A isoform acting on endogenous 5-HT is present in rat major cerebral arteries and is located in nerve endings of fibers arising from dorsal raphe nucleus. PMID- 15352382 TI - Effects of dietary L-arginine supplementation on serum lipids and intestinal enzyme activities in diabetic rats. AB - We investigated whether dietary supplementation with L-arginine, the endogenous precursor of nitric oxide, might affect serum lipid levels and activities of intestinal mucosa enzymes in animals, in which diabetes was induced by administration of streptozotocin. Control and diabetic rats were fed diets with or without 2% L-arginine supplementation for 4 weeks. Diabetic rats had significantly higher concentrations of serum triglycerides and LDL-cholesterol than control rats. These alterations were partially reduced by L-arginine supplementation. Experimental diabetes did not influence the lactase and leucine aminopeptidase activity in the intestine, but the activity of alkaline phosphatase was increased. Furthermore, activities of maltase and sucrase in the intestinal mucosa were elevated in streptozotocin-induced diabetic rats and were restored to control levels after dietary L-arginine supplementation. On the basis of the present experimental evidence, dietary L-arginine supplementation appears to affect the metabolism of lipoproteins and might alleviate some gastrointestinal dysfunctions, commonly seen in diabetes mellitus. PMID- 15352383 TI - Adiposity and dietary intake in cardiovascular risk in an obese population from a Mediterranean area. AB - The aim of the study was to determine the particular relevance of android fat distribution and dietary intake in cardiovascular risk in an obese Mediterranean population with high intake of monounsaturated fatty acids (MUFA) and to compare the findings with those from normal-weight subjects. For the study, 193 subjects aged 25-60 were selected: 118 obese (BMI > or = 27 kg/m2), and 75 normal-weight (BMI < 25 kg/m2). Cardiovascular risk factors including hypertension, dyslipidaemia, glucose intolerance and insulin resistance were assessed. Nutrient intake and body fat distribution were determined. Results show that MUFA were highly consumed in the total population (21% of total energy). The obese population was normolipidemic and normoinsulinemic. However, cardiovascular risk factors (CVRF) were significantly higher than in normal-weight (P < 0.05). Obese subjects derived a greater percentage of their energy intake from total fat and lower from carbohydrates and saturated fats (P < 0.05). BMI and waist-hip ratio positively correlated with fat percentage of total energy intake and with MUFA (g/100 g fatty acids) in men, indicating that the excess of fat intake in obesity is due to a larger consumption of olive oil. CVRF were significantly and positively associated to waist circumference and WHR, both in obese and in normal weight subjects. In conclusion, not only obesity but also android fat in normal weight subjects are important factors in cardiovascular disease even in the Mediterranean population, with a high intake of MUFA, where these factors seem to be more relevant to cardiovascular risk than dietary composition. PMID- 15352384 TI - Influence of fasting on the effects of diazoxide in the ischemic-reperfused rat heart. AB - This investigation aimed to assess whether the mitochondrial ATP-sensitive potassium channel opener diazoxide could reproduce the protection conferred by ischemic preconditioning and to ascertain whether its effects are associated with changes in glycogen breakdown and glycolytic activity. Hearts of fed and 24-h fasted rats were perfused with 10 mM glucose containing medium and exposed to 25 min no-flow ischemia plus 30 min reperfusion. Diazoxide (10 microM) perfusion was begun 10 min before ischemia and continued throughout the experiment. Fasting accelerated reperfusion recovery of contraction, reduced the post-ischemic contracture and decreased lactate accumulation during ischemia but had no effects on glycogen levels and cellular viability. Diazoxide, did not affect glycogen catabolism but improved reperfusion recovery of contraction. Furthermore, diazoxide reduced ischemic lactate accumulation and contracture amplitude only in the fed group whereas it improved cell viability in the fed and fasted groups. These data indicate that: 1) reduced lactate production which may attenuate myocyte acidification might explain, at least in part, the beneficial effects of diazoxide on mechanical function, although data obtained with the fasted rat hearts indicate that other mechanisms must be involved as well; 2) the reduction of lactate production occurring in the fed group, does not seem to be related to glycogenolysis; and 3) since diazoxide improved cell viability in the fasted rat group where it did not reduce glycolytic activity, other mechanisms may be responsible for this cytoprotective effect. PMID- 15352386 TI - Imaging spectrograph for interstellar shocks: a narrowband imaging payload for the far ultraviolet. AB - We present an imaging spectrometer developed for narrowband imaging at 1035 A with high (approximately 1-arc sec) spatial resolution over a modest field of view (approximately 5 arc min). The instrument is based on a conventional Gregorian telescope with aberration-corrected holographic rulings on the secondary optic. These aberration-correcting rulings enable stigmatic imaging in diffracted light with a minimum number of optical elements, thereby maintaining a high system efficiency. The capabilities of this instrument allow us to map the distribution of UV-emitting material in the hot (approximately 300,000 K) plasma from shocks in supernova remnants. Although this design is optimized for imaging near 1035 A, the basic concept can be applied to provide narrowband imaging or long-slit imaging spectroscopy at any wavelength. In addition, a larger field of view is possible with a corresponding loss in spatial resolution. PMID- 15352385 TI - Physiological and metabolic functions of melatonin. AB - Melatonin is a lipophilic hormone, mainly produced and secreted at night by the pineal gland. Melatonin synthesis is under the control of postganglionic sympathetic fibers that innervates the pineal gland. Melatonin acts via high affinity G protein-coupled membrane receptors. To date, three different receptor subtypes have been identified in mammals: MT1 (Mel 1a) and MT2 (Mel 1b) and a putative binding site called MT3. The chronobiotic properties of the hormone for resynchronization of sleep and circadian rhythms disturbances has been demonstrated both in animal models or in clinical trials. Several other physiological effects of melatonin in different peripheral tissues have been described in the past years. In this way, it has been demonstrated that the hormone is involved in the regulation of seasonal reproduction, body weight and energy balance. This contribution has been focused to review some of the physiological functions of melatonin as well as the role of the hormone in the regulation of energy balance and its possible involvement in the development of obesity. PMID- 15352387 TI - Lenticular array for spatial filtering velocimetry of laser speckles from solid surfaces. AB - We present a low-cost optical design for the detection of speckle translation, which can provide measures of in-plane translation or the rotation of a solid structure. A nonspecular target surface is illuminated with coherent light. The scattered light is propagated through an optical arrangement that has been particularly designed for the type of mechanical measurand for which the sensor is intended. The dynamics of the speckle field that arise from the target surface are projected onto a lenticular array, constituting a narrow spatial bandpass filter for the speckle spectrum. The filter provides access to the full phase information of the temporal quasi-sinusoidal intensity output; thus differential arrangements of photodetectors can provide suppression of low-frequency oscillations and higher harmonics, and the direction of the speckle translation can be determined. The spatial filter of the sensor is characterized, and the precision of the sensor when it is integrated with an electronic zero-crossing detection processor is investigated. The best measurement accuracy obtained at constant velocity is 1% at 1.6-mm translation; the relative standard deviation decreases with the square root of the distance traveled. PMID- 15352388 TI - Three-dimensional deformation measurement from the combination of in-plane and out-of-plane electronic speckle pattern interferometers. AB - An optical setup that can be switched to produce in-plane and out-of-plane sensitivity interferometers was designed for three-dimensional deformation measuring by electronic speckle pattern interferometry. Divergent illumination is considered in the evaluation of sensitivity vectors to measure both in-plane and out-of-plane displacement components. The combination of these interferometers presents the advantage of greater sensitivity in directions u, v, and w than a typical interferometer with three illumination beams provides. The system and its basic operation are described, and results with an elastic target that is exposed to a mechanical load are reported. PMID- 15352389 TI - Exact analysis of low-finesse multimode fiber extrinsic Fabry-Perot interferometers. AB - A straightforward theory is presented to accurately model the light inferences in a low-finesse multimode fiber extrinsic Fabry-Perot (FP) interferometer. The effect on the fringe visibility of the gap length, sensor structure imperfections, and modal power distributions is explored. The analysis is particularly useful in the design and optimization of sensors that use an extrinsic FP cavity as the sensing element. PMID- 15352390 TI - Liquid-crystal depolarizer consisting of randomly aligned hybrid orientation domains. AB - A novel depolarization method for linearly polarized incident light that uses a liquid-crystal (LC) cell with randomly aligned hybrid orientation domains is theoretically described by use of Mueller matrix calculations. The depolarization effect of the incident linear polarization is confirmed with Stokes parameter measurements. The unique optical properties of the fabricated LC depolarizer are revealed; that is, the intensity of the transmitted light is independent of the rotation of the analyzer. The degree of polarization becomes zero when the retardation of the LC depolarizer coincides with a half-wavelength. PMID- 15352391 TI - Polarized backlight based on selective total internal reflection at microgrooves. AB - A polarized backlight for LCD illumination is designed and fabricated in which s polarized light is extracted owing to selective total internal reflection at microstructures in the anisotropic layer. From the measurement, the contrast ratio in normal viewing direction can be as high as 64. Luminous uniformity of higher than 80% is achieved for polarized backlights. Furthermore, 1.6 gain in efficiency is obtained aiming for high-efficiency LCD illumination. PMID- 15352392 TI - Use of the delta-P1 approximation for recovery of optical absorption, scattering, and asymmetry coefficients in turbid media. AB - We introduce a robust method to recover optical absorption, reduced scattering, and single-scattering asymmetry coefficients (microa, micro's, g1) of infinite turbid media over a range of (micro's/microa) spanning 3 orders of magnitude. This is accomplished through the spatially resolved measurement of irradiance at source-detector separations spanning 0.25-8 transport mean free paths (l*). These measurements are rapidly processed by a multistaged nonlinear optimization algorithm in which the measured irradiances are compared with predictions given by the delta-P1 variant of the diffusion approximation to the Boltzmann transport equation. The ability of the delta-P1 model to accurately describe radiative transport within media of arbitrary albedo and on spatial scales comparable to l* is the key element enabling the separation of g1 from micro's. PMID- 15352393 TI - Carbon monoxide total column retrievals by use of the measurements of pollution in the troposphere airborne test radiometer. AB - The Measurements of Pollution in the Troposphere (MOPITT) Airborne Test Radiometer (MATR) uses gas correlation filter radiometry from high-altitude aircraft to measure tropospheric carbon monoxide. This radiometer is used in support of the ongoing validation campaign for the MOPITT instrument aboard the Earth Observation System Terra satellite. A recent study of MATR CO retrievals that used data from the autumn of 2001 in the western United States is presented. Retrievals of the CO total column were performed and compared to in situ sampling with less than 10% retrieval error. Effects that influence retrieval, such as instrument sensitivity, retrieval sensitivity, and the bias between observations and the radiative transfer model, are discussed. Comparisons of MATR and MOPITT retrievals show promising consistency. A preliminary interpretation of MATR results is also presented. PMID- 15352394 TI - Design, fabrication, and evaluation of crystal-cored fibers for efficient second harmonic generation based on Cerenkov-radiation-type phase matching. AB - A formulation for calculating the optical second-harmonic power based on the Cerenkov-radiation-type phase matching is derived for a crystal-cored fiber. The prerequisite condition for high efficiency is expressed by a simple relation by use of the refractive indices of a core crystal, a core radius, and a fundamental wavelength. An organic crystal-cored fiber is designed and practically fabricated by the guiding principle derived here. A blue second-harmonic wave of 1.4 mW is obtained from a 1-mm-long fiber by use of a 60-mW semiconductor laser, and the second-harmonic intensity agrees well with the prediction. Degradation of the organic core crystal caused by the generated blue wave is observed, and the lifetime of the device is evaluated. PMID- 15352395 TI - Gated viewing and high-accuracy three-dimensional laser radar. AB - We have developed a fast and high-accuracy three-dimensional (3-D) imaging laser radar that can achieve better than 1-mm range accuracy for half a million pixels in less than 1 s. Our technique is based on range-gating segmentation. We combine the advantages of gated viewing with our new fast technique of 3-D imaging. The system uses a picosecond Q-switched Nd:Yag laser at 532 nm with a 32-kHz pulse repetition frequency (PRF), which triggers an ultrafast camera with a highly sensitive CCD with 582 x 752 pixels. The high range accuracy is achieved with narrow laser pulse widths of approximately 200 ps, a high PRF of 32 kHz, and a high-speed camera with gate times down to 200 ps and delay steps down to 100 ps. The electronics and the software also allow for gated viewing with automatic gain control versus range, whereby foreground backscatter can be suppressed. We describe our technique for the rapid production of high-accuracy 3-D images, derive performance characteristics, and outline future improvements. PMID- 15352396 TI - Ground-based imaging differential optical absorption spectroscopy of atmospheric gases. AB - We describe a compact remote-sensing instrument that permits spatially resolved mapping of atmospheric trace gases by passive differential optical absorption spectroscopy (DOAS) and present our first applications of imaging of the nitrogen dioxide contents of the exhaust plumes of two industrial emitters. DOAS permits the identification and quantification of various gases, e.g., NO2, SO2, and CH2O, from their specific narrowband (differential) absorption structures with high selectivity and sensitivity. With scattered sunlight as the light source, DOAS is used with an imaging spectrometer that is simultaneously acquiring spectral information on the incident light in one spatial dimension (column). The second spatial dimension is scanned by a moving mirror. PMID- 15352397 TI - Characterization of used mineral oil condition by spectroscopic techniques. AB - Optical absorption, fluorescence, and quantitative 13C NMR spectroscopy have been used to study the degradation of mineral gearbox oil. Samples of used oil were collected from field service. Measured absorption, fluorescence, and quantitative 13C NMR spectra of used oils show characteristic changes from the spectra of a fresh oil sample. A clearly observable, approximately 20-nm blueshift of the fluorescence emission occurs during the early stages of oil use and correlates with changes in intensity of some specific 13C NMR resonance lines. These changes correlate with oil age because of the connection between the blueshift and breaking of the larger conjugated hydrocarbons of oil as a result of use. PMID- 15352398 TI - Influence of forward and multiple light scatter on the measurement of beam attenuation in highly scattering marine environments. AB - Using three-dimensional Monte Carlo radiative transfer simulations, we examine the effect of beam transmissometer geometry on the relative error in the measurement of the beam-attenuation coefficient in an aquatic environment characterized by intense light scattering, especially within submerged bubble clouds entrained by surface-wave breaking. We discuss the forward-scattering error associated with the detection of photons scattered at small angles (< 1 degrees) and the multiple-scattering error associated with the detection of photons scattered more than once along the path length of the instrument. Several scattering phase functions describing bubble clouds at different bubble void fractions in the water are considered. Owing to forward-scattering error, a beam attenuation meter (beam transmissometer) with a half-angle of receiver acceptance of 1.0 degrees and a path length of 0.1 m can underestimate the true beam attenuation within the bubble cloud by more than 50%. For bubble clouds with a beam attenuation of as much as 100 m(-1), the multiple-scattering error is no more than a few percent. These results are compared with simulations for some example phase functions that are representative of other scattering regimes found in natural waters. The forward-scattering error for the Petzold phase function of turbid waters is 16% for a typical instrument geometry, whereas for the Henyey Greenstein phase function with the asymmetry parameter of 0.7 and 0.9 the error range is 8-28%. PMID- 15352399 TI - Partially coherent flat-topped beam and its propagation. AB - A partially coherent beam with flat-topped profile is proposed. The cross spectral density of this beam can be expressed as a finite series of the cross spectral density of partially coherent Gaussian-Schell-model beams with different parameters. Analytical propagation formulas for partially coherent flat-topped beams are derived through aligned and misaligned optical systems. The propagation property of partially coherent flat-topped beams in free space is illustrated numerically. The fractional Fourier transform of partially coherent fiat-topped beams is also studied. Our method provides a convenient way to describe partially coherent flat-topped beams and treat their propagation and transformation. PMID- 15352400 TI - Effects of hydrostatic pressure on phase and group modal birefringence in microstructured holey fibers. AB - We calculated the sensitivity of phase (dB/dp) and group (dG/dp) modal birefringence to hydrostatic pressure versus wavelength in two birefringent holey fibers of different construction, where B is the phase modal birefringence, G is the group modal birefringence, and p is the pressure applied to the fiber. The contributions of the geometrical effects that were related only to deformation of the holey structure and the stress-related contribution to the overall pressure sensitivities were analyzed separately. Our results show that these two factors decrease the phase modal birefringence in both structures, which results in negative signs of dB/dp and dG/dp. Furthermore, we demonstrate that the geometrical effects are much weaker than the stress-related effects and contribute only a few percent to the overall pressure sensitivity. PMID- 15352401 TI - Interferometric optical isolator employing a nonreciprocal phase shift operated in a unidirectional magnetic field. AB - An interferometric optical isolator that employs a nonreciprocal phase shift was studied. The optical isolator consisted of an interferometer with distinct layer structures. A traveling light wave underwent distinct nonreciprocal phase shifts such that the optical isolator could be operated in a unidirectional magnetic field. The optical isolator, in which the waveguide had a HfO2 cladding layer in one of the arms, was designed at a wavelength of 1.55 microm. The propagation distance of the nonreciprocal phase shifter required for the isolator's operation was less than 1.5 mm. The device's total length was less than 2 mm. An optical isolator with distinct layer structures was fabricated and evaluated. An isolation ratio of approximately 9.9 dB was obtained in the unidirectional magnetic field. PMID- 15352402 TI - Modeling of energy-transfer upconversion and thermal effects in end-pumped quasi three-level lasers. AB - An analytical model of cw quasi-three-level lasers that includes the influence of energy-transfer upconversion (ETU) has been developed. The results of the general output modeling were applied to a laser with Gaussian beams, and rigorous numerical calculations have been made to study the influence of ETU on threshold, output power, spatial distribution of population-inversion density, and fractional thermal loading. The model was applied to a laser operating at 946 nm in Nd:YAG, where the dependence of laser-beam size on laser performance was investigated in particular. A simple model for the degradation of laser-beam quality from a transversally varying saturated gain is proposed that is in good agreement with measurements of the laser in a plane-plane cavity. PMID- 15352403 TI - Observation and investigation of off-axis modes in a high-power Nd:YAG laser. AB - Unusual transverse patterns were observed in a cw-lamp-pumped Nd:YAG laser near the boundaries of the stability zones of the laser resonator. An interpretation of these patterns is given in terms of off-axis modes, which can be excited owing to spherical aberrations and astigmatism of the thermal lens and thermally induced birefringence. Additional evidence of the off-axis character of the observed modes was obtained by analysis of the temporal behavior of the observed patterns. PMID- 15352404 TI - Push-button medicine. If you can monitor your symptoms electronically, why go to the doctor? PMID- 15352405 TI - What is your diagnosis? Dermatophytic pseudomycetoma. PMID- 15352406 TI - Effect of body size on gastric emptying using the 13C-octanoic acid breath test. AB - The 13C-octanoic acid breath test (OABT) may be a useful non-invasive method for assessing the rate of gastric emptying in dogs. The aim of this study was to determine whether an association exists between body size and rate of gastric emptying in dogs. Fifty-five dogs ranging from 6 to 39 kg were recruited and rate of gastric emptying was assessed using the OABT. The time to peak 13CO2 excretion (tmax) and half-dose recovery time (t1/2) were calculated. The OABT was simple to perform and well tolerated by the dogs. Mean (sd) tmax was 2.67 hours (0.6) and mean t1/2 was 3.38 hours (0.79). Inter-individual variation in the rate of gastric emptying was 23.3 per cent for t1/2 and 22.5 per cent for tmax. No association was detected between the rate of gastric emptying and body surface area, mass, age, sex or test operator. The OABT may be a useful non-invasive and non-radioactive test for assessment of the rate of gastric emptying in dogs in clinical practice. PMID- 15352407 TI - Clinical study of cats injured and killed in road traffic accidents in Cambridgeshire. AB - Data were collected on the injuries, treatment and outcome of 128 cats involved in road accidents and seen as first-opinion cases in veterinary practices in Cambridgeshire. Sixteen cats were dead on arrival; the mortality rate for the remaining cats was 16 per cent. Half of the cats were aged between seven months and two years, with more males than females affected. Most cats had moderate injuries; strays had more severe injuries than owned cats. Areas of the body most often injured were the extremities, head and neck, pelvis and thorax. Skeletal injuries were present in 67 cats and neurological signs in 29. Diagnostic procedures and medical treatment were necessary for the majority of cats; surgery was required in 51 cases. Most cats were hospitalised for between two and seven days and some required up to one month of treatment. The cost of treatment was less than 400 pounds sterling for 84 per cent of cats. PMID- 15352408 TI - Use of ultrasonography to facilitate surgical removal of non-enteric foreign bodies in 17 dogs. AB - Plant material foreign bodies may cause vague clinical signs at the time of initial presentation but can progress to cause persistent or recurrent abscessation, sinus tract formation and debilitating disease, due to migration of the foreign body, severe tissue reaction and secondary infection. Ultrasonography is a non-invasive technique that can be used to identify precisely the presence, location and size of radiolucent foreign bodies. Early identification of foreign bodies using ultrasonography facilitates surgical retrieval before further migration and tissue injury occur. This paper reviews the ultrasonographic findings and outcome in 17 dogs with non-enteric, radiolucent, plant material foreign bodies. PMID- 15352409 TI - Bacteriological study of the liver in dogs. AB - This study aimed to confirm the possible presence of bacteria in the liver of healthy dogs. Laparotomy was performed in 20 animals admitted for routine abdominal surgery. To be selected for the study, dogs had to be healthy adults without clinical liver disease, signs of infection or macroscopic liver abnormalities. Biopsy samples were histologically and bacteriologically examined. Bacteriological analysis was negative for the livers of eight of the dogs. The remaining animals harboured a diverse bacterial flora in their liver. Twelve bacterial species were identified. Histology showed that the livers of 19 dogs had minor or no abnormalities, and only one animal had interstitial fibrosis and trabecular disarrangement. Histological changes were not related to the presence of bacteria. Thus the results showed that the liver of healthy dogs may harbour different bacterial species. These microorganisms did not cause any detectable manifestation of disease, despite being potential pathogens. PMID- 15352410 TI - Traumatic partial hypopituitarism in a cat. AB - Traumatic hypopituitarism was diagnosed in an 11-month-old male neutered cat. The presenting complaints were polydipsia, polyuria and lethargy of three months' duration. Craniocerebral trauma, as a result of a road traffic accident, had preceded the onset of clinical signs by six weeks. Neurological examination revealed right-sided mydriasis, reduced visual and tactile left forelimb placing reflexes and decreased proprioception in both the left fore- and hindlimb. Initial laboratory findings included hypernatraemia, hyperchloraemia, mild azotaemia, eosinophilia and isosthenuria. Low basal cortisol, thyroxine, thyroid stimulating hormone and insulin growth factor-1 were noted. Subsequent to treatment with prednisolone, a water deprivation test confirmed the presence of central diabetes insipidus and therapy with synthetic antidiuretic hormone successfully ameliorated the polydipsia. PMID- 15352411 TI - Massive cystic tetrathyridiosis in a dog. AB - This report describes a case of massive cystic tetrathyridiosis in a four-year old female dobermann. The larvae were distributed in both the peritoneal and pleural cavities, as well as in the lumen of the small intestine along with adult Mesocestoides species. These parasites were identified as tetrathyridia. Numerous cysts were dispersed on the peritoneal, pleural and mesenteric surfaces. The same cysts were seen both on subcapsular surfaces and within sections of the spleen, liver, kidneys and lungs. Histopathological examination also revealed characteristics of tetrathyridium of Mesocestoides species. PMID- 15352412 TI - Post-traumatic paraureteral urinoma in a cat. AB - A retroperitoneal urinoma (uriniferous pseudocyst) was diagnosed in a domestic shorthair cat exhibiting a sublumbar swelling two weeks after a road traffic accident. Plain radiography revealed a soft tissue opacity in the left retroperitoneal space. Intravenous urography and fine-needle aspiration were diagnostic. Contrast was seen pooling in the dilated ipsilateral renal pelvis and proximal ureter. Fluid aspirated from the retroperitoneal space had a creatinine level five times that of serum. Surgical drainage followed by nephrectomy and omentalisation of the pseudocyst was curative. PMID- 15352413 TI - MRI findings in a dog with discospondylitis caused by Bordetella species. AB - A case of discospondylitis in a dog secondary to Bordetella species, diagnosed early with the assistance of magnetic resonance imaging (MRI), is reported. The history and clinical signs were suggestive of possible discospondylitis. MRI identified changes and allowed a presumptive diagnosis of discospondylitis, which was subsequently confirmed by bacterial culture of biopsy material. Discospondylitis associated with Bordetella species infection has not, to the authors' knowledge, been previously reported in the dog. PMID- 15352414 TI - Meningitis in a dog caused by Prevotella oralis. AB - Prevotella oralis, an obligate Gram-negative anaerobe, was shown to be the cause of meningitis in a two-year-old crossbreed dog. The dog had a six-week history of lethargy and neck pain. On presentation, the animal was reluctant to walk, had a stiff gait and neck pain on flexion. Analysis of cerebrospinal fluid documented a neutrophilic pleocytosis (7488 cells/microl) and the presence of occasional intracellular bacterial coccobacilli. The aetiological diagnosis was confirmed by routine aerobic and anaerobic culture, and identification of the isolates. This report also reviews the literature of the documented cases of bacterial meningitis in dogs. PMID- 15352415 TI - Feedback arising from personal experience in practice. PMID- 15352416 TI - Completing the picture: use of CT to investigate elbow dysplasia. PMID- 15352417 TI - Re-establishment of tolerance: the prospect of developing specific treatment for human lupus. PMID- 15352418 TI - The eradication of congenital heart block. PMID- 15352419 TI - Peptides from antibodies to DNA elicit cytokine release from peripheral blood mononuclear cells of patients with systemic lupus erythematosus: relation of cytokine pattern to disease duration. AB - Peptides from VH regions of antibodies to DNA drive immune responses in systemic lupus erythematosus (SLE). We studied peptide-induced cytokine release by peripheral blood mononuclear cells (PBMC) of patients, the influence of peptide concentration, disease characteristics and HLA-D haplotypes. Cells secreting cytokines (IFNgamma, IL-2, IL-4 and IL-10) were measured by ELISPOT in PBMC from 31 patients with SLE and 20 matched healthy controls in response to seven peptides (A-G) from the CDR1/FR2 to CDR2/FR3 VH regions of human anti-DNA MAbs. Disease activity was assessed by SELENA-SLEDAI. HLA-DR and -DQ alleles were determined by molecular typing techniques. PBMC from significantly higher proportions of SLE patients than controls responded to VH peptides by generating IFNgamma and IL-10. Type of cytokines released in response to at least one peptide (D) depended on antigen concentration. Cytokine release was not associated with clinical features of SLE except for disease duration. A shift occurred from IFNgamma, IL-4 and IL-10 production in early disease to IL-4 and IL 10 in late disease (suggesting increasing TH2-like responses over time). Three peptides (B, D, G) were more stimulatory in the SLE patients than controls. Although none of the peptides was restricted by any particular MHC class II allele, among responders there was increased prevalence of HLA- DQB1*0201 and/or DRB1*0301, alleles known to predispose to SLE. Thus, responses to some VH peptides are more frequent in SLE and vary with disease duration. Increased responses in individuals with HLA class II genotypes that predispose to SLE suggest that peptide presentation by those molecules permits brisker peripheral blood cell responses to autoantibody peptides, thus increasing risk for disease. PMID- 15352420 TI - Headaches in patients with systemic lupus erythematosus: a comparative study. AB - The incidence and nature of headaches in 85 systemic lupus erythematosus (SLE) patients attending an outpatient clinic were studied and compared to those experienced by 61 nurses. The two groups were similar in age, sex and ethnicity. Test-retest assessment of reliability gave both groups 95% confidence limits of 0.09-0.21. Thirty-two (38%) patients developed migrainous headaches and nine (10%) stress headaches with the onset of lupus. In the control group, four (6%) developed migraine and 40 (66%) developed stress headaches on commencing work. We could not document any association of headaches with flares of systemic disease, the ACA syndrome, Raynaud's phenomenon or increased SLEDAI score. We conclude that migrainous headaches are more common in lupus patients than healthy controls, but in an outpatient setting are not statistically associated with flares of systemic disease. PMID- 15352421 TI - Pulmonary hypertension in systemic lupus. AB - Pulmonary arterial hypertension (PAH) has devastating consequences in the rheumatic diseases; however, the prevalence in lupus is not well delineated. We searched the University of Toronto lupus database to ascertain the first echocardiogram ordered at their physician's discretion between 1995 and 2002. We reviewed the echocardiogram reports for right ventricular systolic pressure (RVSP), valvular disease, and atrial and ventricular function. The PAH was defined as RVSP > or = 40 mmHg. Patients were divided into three groups: RVSP > or = 40 mmHg, RVSP = 30-39 mmHg and RVSP < 30 mmHg. We analysed potential associations between presence of PAH and lupus including disease activity, organ involvement and anticardiolipin antibodies, both at the time of and any time prior to echocardiography. In total, 129 patients underwent echocardiography. Nine patients' echocardiograms were not obtainable, and three patients were excluded from analysis, as their visit was more than six months from the date of echocardiography. Sixteen patients (14%) had RVSP > or = 40 mmHg, 43 (37%) patients had RVSP of 30-39, and 60 (51%) patients had RVSP < 30 mmHg. There was no statistical difference in disease activity, organ involvement or serology among all three groups. In conclusion, the prevalence of PAH (RVSP > or = 40 mmHg) on first echocardiogram ordered at physician discretion in our cohort was 14%. An RVSP of 30-39 mmHg was found in 37% of patients. Although abnormal, the clinical significance of this finding is unknown. Disease activity, organ involvement and anticardiolipin antibodies were not associated with PAH. Further research is needed to identify the mechanism, response to immunosuppression and impact on quality of life in these patients. PMID- 15352422 TI - A putative regulatory polymorphism in PD-1 is associated with nephropathy in a population-based cohort of systemic lupus erythematosus patients. AB - The association between polymorphisms in the programmed death (PD-1) gene and susceptibility to systemic lupus erythematosus (SLE) was determined using genomic DNA, isolated from a population-based cohort of 95 SLE patients and 155 healthy controls. Polymorphisms in the complete PD-1 gene except the large intron 1 were detected by sequencing. Furthermore, the patients were stratified according to the presence or absence of lupus nephropathy. The influence of the detected single nuclear polymorphisms (SNPs) on this specific clinical disease parameter was determined. In total, we identified 12 single nucleotide polymorphisms, of which six were novel and eight were considered to be rare (the frequency of the minor allele of these was less than 1% in our study populations). We found a significant association of an intronic 6867C/G SNP in the PD-1 gene with the presence of lupus nephropathy. As the 6867C/G SNP is located in a putative binding site for the transcriptional repressor ZEB, the associated allele of this SNP potentially alters the transcriptional regulation of PD-1. This report, for the first time, indicates that a 6867C/G SNP of the PD-1 gene is associated with lupus nephropathy in Caucasian SLE patients. PMID- 15352423 TI - Reduced erythrocyte complement receptor type 1 in systemic lupus erythematosus is related to a disease activity index and not to the presence or severity of renal disease. AB - The present study investigated the expression of the complement receptor type 1 (CR1) on the membrane of erythrocytes (CR1/E) of patients with systemic lupus erythematosus (SLE) by flow cytometry. We found a significant reduction in CR1/E numbers in SLE patients (n = 52), compared to controls (512 +/- 171 and 689 +/- 146, respectively, P = 0.0001). Reduction was more pronounced in active disease patients. The mean CR1/E number observed in patients with inactive disease was 546 +/- 163 CR1/E, while active SLE patients presented a mean of 385 +/- 133 CR1/E (P = 0.001). Patients with SLE with similar activity indexes tend to have similar CR1/E numbers, irrespective of disease severity. We also observed a trend to CR1/E reduction in severe nephritis patients. A small group of SLE patients with chronic renal failure and inactive disease showed CR1/E numbers nearly identical to controls (689 +/- 146 versus 686 +/- 123, respectively, P = 0.95). This was the only group of SLE patients with normal CR1/E numbers. These results confirm the CR1/E reduction in SLE patients as previously described, and also suggest that this reduction is related to disease activity and not to disease severity. PMID- 15352424 TI - Antibodies to mannose binding lectin in patients with systemic lupus erythematosus. AB - Deficiency of mannose binding lectin (MBL), a C-type lectin with structural similarities to C1q, has been shown to predispose to the development of systemic lupus erythematosus (SLE). Some patients have low serum MBL levels which cannot be explained by either structural gene mutations or promoter polymorphisms. The objective of this study was to detect the presence of autoantibodies against MBL and to evaluate their relationship to serum MBL levels. Anti-MBL antibodies of IgM and IgG classes from consecutive SLE patients (n = 135) and healthy subjects (n = 50) were measured by an in-house ELISA. Using the 90th percentile of controls as a cutoff, more SLE patients [23.7% (32/135)] were found to have IgG anti-MBL antibodies than normal controls [10.0% (5/50)] (P = 0.04). The same trend was observed when ethnicity was taken into account by analysing Caucasians alone (n = 90). IgM anti-MBL antibodies were only found in two SLE patients (2/22, 9.1%) who had no concomitant IgG anti-MBL antibodies. Serum levels of IgG anti-MBL antibodies were found to correlate with serum MBL levels (r = 0.55, P = 0.049). However, the levels of anti-MBL antibodies did not correlate with overall disease activity. Thus the production of anti-MBL antibodies is likely to be a specific antigen-driven process. Its role in lupus pathogenesis remains to be elucidated. PMID- 15352425 TI - Health care and burden of illness in systemic lupus erythematosus compared to rheumatoid arthritis: results from the National Database of the German Collaborative Arthritis Centres. AB - During the past 20 years, outcome studies in the rheumatic diseases have, on the one hand, given increasing evidence of the unfavourable long-term prognosis of rheumatoid arthritis (RA) and on the other hand determined continuous improvement of prognosis in systemic lupus erythematosus (SLE). The aim of the study was to investigate how this translates into the current spectrum of patients with rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) seen by rheumatologists in Germany and to compare aspects of the burden of disease, disease outcomes and treatment between these two important rheumatic diseases using a large clinical database. Current health care was analysed with data from the German rheumatological database of 10 068 patients with RA and 1248 patients with SLE seen by rheumatologists in 2001. In addition, of a total of 3546 patients with SLE and 24 969 patients with RA seen at the German Collaborative Arthritis Centres between 1994 and 2001, 3465 cases of SLE were matched by age, sex, disease duration and referral status with a corresponding RA case. There were considerable differences in treatment of patients before referral to a rheumatologist and in rheumatologic care. In 2001, patients with SLE were treated by their rheumatologists mainly with antimalarials (AM, 37%), azathioprine (29%) and nonselective NSAIDs (16%). Of them, 61% received at least one immunosuppressive drug (including AM) plus glucocorticoids. In RA, methotrexate was the predominant medication (63%), and 56% received at least one immunosuppressive drug plus glucocorticoids. Matched pairs analysis showed that SLE patients with a short disease duration were almost equally burdened by pain, functional limitations and reduced health status as RA patients. After a disease duration of >10 years, however, patients with RA showed poorer outcomes than those with SLE: RA patients reported significantly more often severe pain (30% in RA versus 17% in SLE) and poor global health status (52 versus 38%), and their disease activity as well as severity was rated higher by the rheumatologists. In conclusion, comparing large groups of RA and SLE patients we found a similar burden in early but not in late disease. Taking into account limitations as to the generalizability of the results (recruitment in rheumatologic care, cross sectional data, underestimation of SLE-specific outcomes), the discrepancy between the high increase in disease-related negative outcomes with longer disease duration in RA but not in SLE indicates a better long-term prognosis in SLE concerning the items observed. The great disparity in treatment intensity between rheumatologists and nonrheumatologists shows that the involvement of a specialist is needed equally in both diseases. PMID- 15352426 TI - Outcome of patients with systemic lupus erythematosus in intensive care unit. AB - The objective of the study was to identify the causes, outcome and prognosis of severe illness in patients with systemic lupus erythematosus (SLE) requiring intensive care unit (ICU) care in a University Hospital over a five-year period. The design was a cohort study. Forty-eight SLE patients requiring ICU management over a five-year period (January 1997-December 2001) were studied prospectively. Of 48 patients, 14 (29.2%) died, predominantly with multiorgan dysfunction syndrome (MODS). Patients whose APACHE II score was equal to or greater than 20 had higher mortality than those with APACHE score below 20 (60 versus 7.1%; and P < 0.01). All the 18 patients whose health status rated as 'good' survived, while 46.7% of 30 patients whose health rated as 'poor' died (P < 0.01). Patients who had thrombocytopenia associated with sepsis and/or disseminated intravascular coagulopathy (DIC) had the highest mortality (75%, five-year survival). In conclusion, SLE patients admitted to the ICU had a lower mortality rate than some of the previous reports. Patients with SLE with high APACHE score, > or =20, poor health status, thrombocytopenia and multiorgan dysfunction syndrome had poor prognosis in the ICU. PMID- 15352427 TI - First international meeting on mycophenolate mofetil: an advance in the treatment of lupus 27 April 2004, St Thomas' Hospital, London, UK. PMID- 15352428 TI - A case of systemic lupus erythematosus with giant hepatic cavernous hemangioma. AB - A 39-year old woman with antiphospholipid antibody positive systemic lupus erythematosus (SLE) was admitted to our department because of high fever, liver dysfunction and high level of C-reactive protein. At hospitalization, there was no anemia or jaundice. A tumor was palpable in the epigastric region, and there was tenderness in this region, but no muscular defense. There were no findings which indicated disease activity of SLE. The result of abdominal ultrasonography showed that there was a giant tumor, which occupied the majority of the left lobule of the liver, and a nonuniform ultrasound image was observed inside the tumor. The result of dynamic computed tomography (CT) showed peripheral globular enhancement, and enhancement then extended to the tumor center with time. Consequently, the patient was diagnosed with multiple hepatic hemangiomas. After admission, anemia rapidly deteriorated, and platelet count tended to decline. Therefore, intratumor hemorrhage was suspected, and emergent angiography was performed. For hemostatic purposes, transcatheter arterial embolization (TAE) and extended left hepatic lobectomy were performed. In patients with autoimmune diseases such as SLE and antiphospholipid syndromes, when thrombocytopenia is observed, care should be paid to identifying its cause, considering thrombocytopenia may be induced by hemangioma, although these cases are extremely rare. PMID- 15352429 TI - Lupus pneumonitis or severe acute respiratory syndrome? AB - Severe acute respiratory syndrome (SARS) is a serious respiratory illness caused by a novel human coronavirus. The disease is highly infectious and carries significant mortality and morbidity. There was a major outbreak of SARS in Guangdong, Taiwan, Beijing, Hong Kong and Toronto between March and June 2003. Common presenting features of SARS are high fever, chills, rigor, malaise, nonproductive cough, lymphopenia and pulmonary infiltrates, followed by rapidly progressive respiratory failure in some cases. We describe two patients with systemic lupus erythematosus (SLE) who presented with fever, systemic upset and pulmonary infiltrates between April and June, 2003. One patient was confirmed to have coronavirus pneumonia while the other had active SLE with lung involvement. Our cases illustrate the difficult diagnostic dilemma in the evaluation of febrile SLE patients during the SARS epidemic. PMID- 15352430 TI - Therapeutic management of antiphospholipid syndrome in aspirin sensitive patients. PMID- 15352431 TI - Tubular aggregates in a patient with primary antiphospholipid syndrome. PMID- 15352432 TI - No water, no future. PMID- 15352433 TI - Visualizing contaminants in plants. PMID- 15352434 TI - Indoor mold standard could prove elusive. PMID- 15352435 TI - Airborne mycotoxins discovered in moldy buildings. PMID- 15352436 TI - Rapid indicators of beach pollution needed. PMID- 15352437 TI - Florida, EPA slammed for regulatory failures. PMID- 15352438 TI - Untangling the causes of coral reef decline. PMID- 15352439 TI - Why carbon monoxide still matters. PMID- 15352440 TI - Mycotoxin risk assessment for the purpose of setting international regulatory standards. AB - The 2003 Council for Agricultural Science and Technology Mycotoxin report states that one 21st century goal is the development of uniform regulations worldwide for foodborne mycotoxin contamination. This study informs that endeavor by a risk assessment and economic analysis of two important mycotoxins: fumonisins and aflatoxins. The goals are to identify the nations that would be most heavily impacted by tighter mycotoxin regulations, examine costs and benefits as a function of regulatory stringency, and address risk-risk tradeoffs between health benefits and economic losses from compliance with those regulations. Among industrial nations, the United States would experience the heaviest economic losses from more precautionary mycotoxin standards. Environmental conditions in the developing world, however, are more conducive to mycotoxin accumulation in crops. Contrary to concerns expressed among policymakers, the less developed countries that would likely experience the greatest loss from tighter mycotoxin standards are not sub-Saharan African nations, but China and Argentina. If a fumonisin standard of 0.5 mg/kg were adopted worldwide, total export losses from fumonisins in corn may exceed 300 million dollars annually: 3-fold higher than if the less stringent U.S. standard of 2 mg/kg were adopted. Likewise, export losses from aflatoxins in peanuts may exceed 450 million dollars under the current EU regulatory standard of 4 microg/kg: almost 5-fold higher than if the U.S. standard of 20 microg/kg were adopted. Stricter standards are unlikely to improve health significantly. In developing nations such as China where hepatitis B and C are prevalent, tighter aflatoxin standards may increase health risks until improved control methods for aflatoxins are found, as high-quality crops may be exported instead of being consumed domestically. PMID- 15352441 TI - Perfluorinated compounds in coastal waters of Hong Kong, South China, and Korea. AB - Perfluorinated compounds (PFCs), such as perfluorooctanesulfonate (PFOS) and related compounds, have recently been identified in the environment. PFOS, the terminal degradation product of many of the PFCs, has been found globally in many wildlife species, as well as open ocean waters, even in remote regions far from sources. In this study, a solid-phase extraction procedure coupled with high performance liquid chromatography interfaced to high-resolution mass spectrometry was used to isolate, identify, and quantify small concentrations of PFCs in seawater. These techniques were applied to investigate the local sources of PFCs in several industrialized areas of Asia and provide information on how the PFCs are circulated by coastal currents. Ranges of concentrations of PFOS in coastal seawaters of Hong Kong, the Pearl River Delta, including the South China Sea, and Korea were 0.09-3.1, 0.02-12, and 0.04-730 pg/mL, respectively, while those of perfluorooctanoic acid (PFOA) were 0.73-5.5, 0.24-16, and 0.24-320 pg/mL, respectively. Potential sources of PFCs include major industrialized areas along the Pearl River Delta of southern China and major cities of Korea, which are several of the fastest growing industrial and economic regions in the world. Detectable concentrations of PFOS and PFOA in waters of southern China were similar to those in the coastal marine environment of Japan and certain regions in Korea. Concentrations of PFCs in several locations in Korean waters were 10 100-fold greater than those in the other locations on which we report here. The spatial and seasonal variations in PFC concentrations in surface seawaters in the Pearl River Delta and South China Sea indicate the strong influence of the Pearl River discharge on the magnitude and extent of PFC contamination in southern China. All of the concentrations of PFOS were less than those that would be expected to cause adverse effects to aquatic organisms or their predators except for one location in Korea adjacent to an industrialized area. Hazard quotients were from <0.001 to 0.002 for aquatic animals and ranged from <0.001 to 17 for predatory birds. PMID- 15352442 TI - Detection of perfluorooctane surfactants in Great Lakes water. AB - Widespread use of perfluorooctane surfactants has led to ubiquitous presence of these chemicals in biological tissues. While perfluorooctane surfactants have been measured in blood and liver tissue samples of fish, birds, and mammals in the Great Lakes region, data for the aqueous concentrations of these compounds in the Great Lakes or other ambient waters is lacking. Sixteen Great Lakes water samples were analyzed for eight perfluorooctane surfactants. The monitored perfluorooctane surfactants were quantitatively determined using single quadrupole HPLC/MS and qualitatively confirmed using ion trap MS/MS. Additionally, PFOS was quantitatively confirmed using triple quadrupole LC/MS/MS. Concentrations of PFOS and PFOA in the two lakes ranged from 21-70 and 27-50 ng/L, respectively. Analysis also showed the presence of PFOS precursors, N EtFOSAA (range of 4.2-11 ng/L) and FOSA (range of 0.6-1.3 ng/L), in all samples above the LOQ. PFOSulfinate, another precursor, was identified at six of eight locations with a concentration range, when present, of <2.2-17 ng/L. Other PFOS precursors, N-EtFOSE, PFOSAA, and N-EtFOSA were not observed at any of the sampling locations. These are the first reported concentrations of perfluorooctane surfactants in Great Lakes water and the first report of PFOS precursors in any water body. PMID- 15352443 TI - Brominated organic contaminants in the liver and egg of the common cormorants (Phalacrocorax carbo) from Japan. AB - The contamination profiles of brominated flame retardants (BFRs) such as polybrominated diphenyl ethers (PBDEs), biphenyls (PBBs), dibenzo-p-dioxins (PBDDs), and dibenzofurans (PBDFs) were determined in the liver and egg of common cormorants from Japan. PBDEs and PBBs were detected in all the samples; especially the former were detected at elevated levels. PBDDs/PBDFs were also detected in cormorants, albeit the concentrations were lower than those of the PBBs. The total concentration of PBDEs ranged from 330 to 2600 in the liver and from 600 to 3300 in the egg on a nanogram per gram of lipid basis. The concentration of PBBs was in the range from 3.0 to 33 (in the liver) and from 3.4 to 82 (in the egg) on a nanogram per gram of lipid basis. The 2,2',4,4',5,5' hexabromobiphenyl (BB-153) was the most predominant PBB congener in either organ, which corresponds to a major constituent of the BFR FireMaster BP-6. Concentrations of PBDDs/ PBDFs in the liver (range from 21 to 470) were slightly higher than in the egg (range from 31 to 160) on a picogram per gram of lipid basis. The results of this study imply that common cormorants accumulate a high level of PBDEs and PBBs. Comparing the concentrations of brominated organic compounds with those of chlorinated analogues, good relevance between PBBs and coplanar PCBs (r2 = 0.746 [liver] and 0.715 [egg]) was elucidated. To our knowledge, the present study demonstrates the first report of PBDEs, PBBs, and PBDDs/PBDFs in the common cormorant from Japan. PMID- 15352444 TI - Sorption of diverse organic vapors to snow. AB - Sorption from air to one snow sample has been measured for a broad set of organic vapors covering a wide range of physicochemical properties. Those results that could be compared to literature values mostly lay in the same order of magnitude. As expected, a fit with the vapor pressure did not reveal a good correlation (R2 = 0.11). Therefore, the data set was interpreted with a linear free energy relationship, based on intermolecular interactions (van der Waals interactions and hydrogen bond interactions). Although we cannot assign the observed sorption to a specific process (adsorption to the snow crystal surface, incorporation in the solid ice crystal, absorption into a quasi-liquid layer, or grain boundary effects), the model provides a useful tool for the prediction of snow sorption for other compounds: log K(i snow suface/air) = 0.639 (+/- 0.037) log K(i hexadecane/air) + 3.38 (+/- 0.17) sigmabetai + 3.53 (+/- 0.25) sigmaalphai - 6.85. The sorption coefficients measured could be described well with the compound parameters used (subscript i), with an R2 = 0.90. PMID- 15352445 TI - Historical record of European emissions of heavy metals to the atmosphere since the 1650s from alpine snow/ice cores drilled near Monte Rosa. AB - Cr, Cu, Zn, Co, Ni, Mo, Rh, Pd, Ag, Cd, Sb, Pt, Au, and U have been determined in clean room conditions by inductively coupled plasma sector field mass spectrometry and other analytical techniques, in various sections of two dated snow/ice cores from the high-altitude (4450 m asl) glacier saddle Colle Gnifetti, Monte Rosa massif, located in the Swiss-Italian Alps. These cores cover a 350 year time period, from 1650 to 1994. The results show highly enhanced concentrations for most metals in snow/ice dated from the second half of the 20th century, compared with concentrations in ancient ice dated from the 17th and 18th centuries. The highest increase factors from the pre-1700 period to the post-1970 period are observed for Cd (36), Zn (19), Bi (15), Cu (11), and Ni (9), confirming the importance of atmospheric pollution by heavy metals in Europe. Metal concentrations observed in Colle Gnifetti snow around 1980 appear to be quantitatively related to metal emissions from Italy, Switzerland, Germany, France, Belgium, and Austria at that time, making it possible to reconstruct past changes in metal emission in these countries during the last centuries. PMID- 15352446 TI - Are brook trout streams in western Virginia and Shenandoah National Park recovering from acidification? AB - Streamwater composition data obtained through periodic sampling of streams that support brook trout (Salvelinus fontinalis) in the mountains of western Virginia were examined for evidence of recovery from acidification during the 1988-2001 period. Measurements of sulfate deposition in precipitation indicate that sulfate deposition in the region declined approximately 40% between 1985 and 2000. While no significant regional trends in acid-base constituents were observed for the set (n = 65) of western Virginia study streams, significant regional trends were observed for a subset (n = 14) of streams in Shenandoah National Park (SNP). For the subset of SNP streams, the median increase in acid-neutralizing capacity (ANC) was 0.168 microequiv L(-1) year(-1) and the median decrease in sulfate concentration was -0.229 microequiv L(-1) year(-1). Although these trends are consistent with recovery from acidification, the degree of apparent recovery is small compared to estimates of historic acidification in SNP streams and much less than observed in other, more northern regions in the United States. Correlation between sulfate concentration trends and current sulfate concentrations in streamwater suggests that recovery from stream acidification in the western Virginia region is determined by sulfur retention processes in watershed soils. A transient increase in nitrate concentrations that occurred among some western Virginia streams following forest defoliation by the gypsy moth (Lymantria dispar) complicates interpretation of the observed patterns of change in acid-base status. PMID- 15352447 TI - Investigating the molecular interactions of oxytetracycline in clay and organic matter: insights on factors affecting its mobility in soil. AB - The interactions of oxytetracycline with model clay adsorbents were investigated as a function of suspension pH. The clay adsorbents used were native montmorillonite (SWy-2), Na-montmorillonite (Na-SWy-2), and hexadecyl trimethylammonium-montmorillonite (HDTMA-montmorillonite). The adsorption of oxytetracycline to the clay could be described by Freundlich-type adsorption isotherms. It was observed that the adsorption of oxytetracycline in the native and sodium forms of montmorillonite decreases with increasing pH in the order pH 1.5 > 5.0 > 8.7 > 11.0. This trend is consistent with cationic exchange interactions that are dominant at lower pH values when oxytetracycline has a net positive charge. On the other hand, hydrophobic interactions when oxytetracycline is zwitterionic (at pH 5.0) are predominant over other mechanisms, as evident from the FT-IR spectrum of the HDTMA-montmorillonite and humic acid montmorillonite adsorbed with oxytetracycline. The presence of a large amount of dissolved organic matter (DOM) was also found to decrease the sorption of oxytetracycline to clay, suggesting that DOM may increase its mobility in the natural environment. Several mechanisms of interaction of oxytetracycline in clay are proposed based on the adsorption isotherms and the results from X-ray diffraction (XRD) and Fourier transform-infrared (FT-IR) analyses. PMID- 15352448 TI - Biotransformation of tributyltin to tin in freshwater river-bed sediments contaminated by an organotin release. AB - The largest documented release of organotin compounds to a freshwater river system in the United States occurred in early 2000 in central South Carolina. The release consisted of an unknown volume of various organotin compounds such tetrabutyltin (TTBT), tributyltin (TBT), tetraoctyltin (TTOT), and trioctyl tin (TOT) and resulted in a massive fish kill and the permanent closures of a municipal wastewater treatment plant and a local city's only drinking-water intake. Initial sampling events in 2000 and 2001 indicated that concentrations of the ecologically toxic TTBT and TBT were each greater than 10 000 microg/kg in surface-water bed sediments in depositional areas, such as lakes and beaver ponds downstream of the release. Bed-sediment samples collected between 2001 and 2003, however, revealed a substantial decrease in bed-sediment organotin concentrations and an increase in concentrations of degradation intermediate compounds. For example, in bed sediments of a representative beaver pond located about 1.6 km downstream of the release, total organotin concentrations [the sum of TTBT, TBT, and the TBT degradation intermediates dibutyltin (DBT) and monobutyltin (MBT)] decreased from 38 670 to 298 microg/kg. In Crystal Lake, a large lake about 0.4 km downstream from the beaver pond, total organotin concentrations decreased from 28 300 to less than 5 microg/kg during the same time period. Moreover, bed sediment inorganic tin concentrations increased from pre-release levels of less than 800 to 32 700 microg/kg during this time. These field data suggest that the released organotin compounds, such as TBT, are being transformed into inorganic tin by bed-sediment microbial processes. Microcosms were created in the laboratory that contained bed sediment from the two sites and were amended with tributyltin (as tributyltin chloride) under an ambient air headspace and sacrificially analyzed periodically for TBT, the biodegradation intermediates DBT and MBT, and tin. TBT concentrations decreased faster [half-life (t1/2) = 28 d] in the organic-rich sediments (21.5%) that characterized the beaver pond as compared to the slower (t1/2 = 78 d) degradation rate in the sandy, organic-poor, sediments (0.43%) of Crystal Lake. Moreover, the concentration of inorganic tin increased in microcosms containing bed sediments from both locations. These field and laboratory results suggest that biotransformation of the released organotins, in particular the ecologically detrimental TBT, does occur in this fresh surface water system impacted with high concentrations of neat organotin compounds. PMID- 15352449 TI - Photooxidation and its effects on the carboxyl content of dissolved organic matter in two coastal rivers in the southeastern United States. AB - Photodecarboxylation (often stoichiometrically expressed as RCOOH + (1/2)O2 --> ROH + CO2) has long been postulated to be principally responsible for generating CO2 from photooxidation of dissolved organic matter (DOM). In this study, the quantitative relationships were investigated among O2 consumption, CO2 production, and variation of carboxyl content resulting from photooxidation of DOM in natural water samples obtained from the freshwater reaches of the Satilla River and Altamaha River in the southeastern United States. In terms of loss of dissolved organic carbon (DOC), loss of optical absorbance, and production of CO2, the rate of photooxidation of DOM was increased in the presence of Fe redox chemistry and with increasing O2 content. The ratio of photochemical O2 consumption to CO2 photoproduction ranged from approximately 0.8 to 2.5, depending on the O2 content, the extent of involvement of Fe, and probably the initial oxidation state of DOM as well. The absolute concentration of carboxyl groups ([-COOH]) on DOM only slightly decreased or increased over the course of irradiation, possibly depending on the stages of photooxidation, while the DOC normalized carboxyl content substantially increased in the presence of Fe redox chemistry and sufficient O2. Both the initial [-COOH] and the apparent loss of this quantity over the course of irradiation was too small to account for the much larger production of CO2, suggesting that carboxyl groups were photochemically regenerated or that the major production pathway for CO2 did not involve photodecarboxylation. The results from this study can be chemically rationalized by a reaction scheme of (a) photodecarboxylation/ regeneration of carboxyl: CxHyOz(COOH)m + aO2 + (metals, hv) --> bCO2 + cH2O2 + Cx-bHy'Oz'(COOH)m b(COOH)b or of (b) nondecarboxylation photooxidation: CxHyOz(COOH)m + aO2 + (metals, hv) --> bCO2 + cH2O2 + Cx-bHy'Oz'(COOH)m. PMID- 15352450 TI - Hydration of natural organic matter: effect on sorption of organic compounds by humin and humic acid fractions vs original peat material. AB - Natural organic matter (NOM) hydration is found to change activity-based sorption of test organic compounds by as much as 2-3 orders of magnitude, depending on the compound and the specific NOM sorbent. This is demonstrated for sorption on humin, humic acid, and the NOM source material. Hydration assistance in organic compound sorption correlates with the ability of the sorbate to interact strongly with hydrated sorbents, demonstrating the important role of noncovalent polar links in organizing the sorbent structure. Differences in hydration effect between the sorbents are caused mainly by differences in compound-sorbent interactions in the dry state. For a given compound, hydration of the sorbent tends to equalize the sorption capability of the three sorbents. No correlation was found between the strength of sorbate-sorbent interactions or the type of sorbate functional groups and the extent of sorption nonlinearity. Sorption nonlinearity compared over the same sorbed concentration range is greater on the original NOM than on either of the two extracted fractions. In elucidating sorption mechanisms on hydrated NOM, it is important to explicitly consider the participation of water molecules in organic compound interactions in the NOM phase. PMID- 15352451 TI - Biodegradation of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) by Phanerochaete chrysosporium: new insight into the degradation pathway. AB - Octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) is a recalcitrant energetic chemical that tends to accumulate in soil, close to the surface. The present study describes the aerobic biodegradability of HMX using Phanerochaete chrysosporium. When added to 7 day old static P. chrysosporium liquid cultures, HMX (600 nmol) degraded within 25 days of incubation. The removal of HMX was concomitant with the formation of transient amounts of its mono-nitroso derivative (1-NO-HMX). The latter apparently degraded via two potential routes: the first involved N-denitration followed by hydrolytic ring cleavage, and the second involved alpha-hydroxylation prior to ring cleavage. The degradation of 1 NO-HMX gave the ring-cleavage product 4-nitro-2,4-diazabutanal (NDAB), nitrite (NO2 -), nitrous oxide (N2O), and formaldehyde (HCHO). Using [14C]-HMX, we obtained 14CO2 (70% in 50 days), representing three C atoms of HMX. Incubation of real soils, contaminated with either HMX (403 micromol kg(-1)) (military base soil) or HMX (3057 micromol kg(-1)), and RDX (342 micromol kg(-1)) (ammunition soil) with the fungus led to 75 and 19.8% mineralization of HMX (liberated 14CO2), respectively, also via the intermediary formation of 1-NO-HMX. Mineralization in the latter soil increased to 35% after the addition of glucose, indicating that a fungus-based remediation process for heavily contaminated soils is promising. The present findings improve our understanding about the degradation pathway of HMX and demonstrate the utility of using the robust and versatile fungus P. chrysosporium to develop effective remediation processes for the removal of HMX. PMID- 15352452 TI - Odorous products of the chlorination of phenylalanine in water: formation, evolution, and quantification. AB - To explain some of the possible origins of an odor episode which took place in a drinking water supply in the region of Paris (France), the chlorination reaction in water of phenylalanine was studied. This amino acid was chosen for first experiments because of its physical and chemical particular properties. Changes in the different byproducts formed were followed by high-performance liquid chromatography (HPLC) over a period of time. N-chlorophenylalanine (mono-N chlorinated amino acid) and then phenylacetaldehyde were the major products formed for the lower chlorine to nitrogen molar ratios. For Cl/N molar ratios of 1 and beyond, phenylacetonitrile and N-chlorophenylacetaldimine appeared and increased with the chlorination level. N-chlorophenylacetaldimine was quantified by using its difference of stability in various organic solvents. Our attention was first directed to the monochlorinated derivative but further examination indicated that it could not be responsible for odor troubles: it dissociated before reaching the consumer's tap and it was produced at consistently low yields under conditions relevant to drinking water treatment. On the contrary, chloroaldimine appeared to be a very odorous and water-stable product: it strongly smells of swimming pool with a floral background. The odor detection threshold is about 3 microg x L(-1) and it can persist for more than one week at 18 degrees C. It is now suspected of being a source of off-flavor concerns among consumers. PMID- 15352453 TI - Do arsenosugars pose a risk to human health? The comparative toxicities of a trivalent and pentavalent arsenosugar. AB - Seafood frequently contains high concentrations of arsenic (approximately 10-100 mg/kg dry weight). In marine algae (seaweed), this arsenic occurs predominantly as ribose derivatives known collectively as arsenosugars. Although it is clear that arsenosugars are not acutely toxic, there is a possibility of arsenosugars having slight chronic toxicity. In general, trivalent arsenicals are more toxic than their pentavalent counterparts, so in this work we examine the hypothesis that trivalent arsenosugars might be significantly more toxic than pentavalent arsenosugars in vitro. We compared the in vitro toxicity of (R)-2,3 dihydroxypropyl-5-deoxy-5-dimethylarsinoyl-beta-D-riboside, a pentavalent arsenosugar, to that of its trivalent counterpart, (R)-2,3-dihydroxypropyl-5 deoxy-5-dimethylarsino-beta-D-riboside. The trivalent arsenosugar nicked plasmid DNA, whereas the pentavalent arsenosugar did not. The trivalent arsenosugar was more cytotoxic (IC50 = 200 microM, 48 h exposure) than its pentavalent counterpart (IC50 > 6000 microM, 48 h exposure) in normal human epidermal keratinocytes in vitro as determined via the neutral red uptake assay. However, both the trivalent and the pentavalent arsenosugars were significantly less toxic than MMA(III), DMA(III), and arsenate. Neither the pentavalent arsenosugar nor the trivalent arsenosugar were mutagenic in Salmonella TA104. The trivalent arsenosugar was readily formed by reaction of the pentavalent arsenosugar with thiol compounds, including, cysteine, glutathione, and dithioerythritol. This work suggests that the reduction of pentavalent arsenosugars to trivalent arsenosugars in biology might have environmental consequences, especially because seaweed consumption is a significant environmental source for human exposure to arsenicals. PMID- 15352454 TI - Solar photodecomposition of decabromodiphenyl ether: products and quantum yield. AB - Decabromodiphenyl ether (BDE209) is a widely used flame retardant, yet information regarding its environmental transformation rates and pathways are largely unknown. Because photochemical transformation is often suggested as a potentially important fate process for BDE209, the reaction rate and products of the solar degradation under favorable solvent conditions were determined in this study. Decabromodiphenyl ether (BDE209), dissolved in hexane, reacts in minutes via direct solar irradiation, at midlatitude (40 degrees 29' N, 86 degrees 59.5' W) in afternoon July and October sunlight. Observed first-order reaction rate constants, kobs, at the different exposure times were kobs = 1.86 x 10(-3) s(-1) (July) and kobs = 1.11 x 10(-3) s(-1) (October). The photodecomposition quantum yield was calculated from these data and from the solar irradiance data measured at 300, 305.5, 311.4, 317.6, 325.4, 332.4, and 368 nm reported at a USGS UVB monitoring station located nearby. The range of wavelengths where both the molar absorptivity of BDE209 and the solar irradiance flux are significant occurs between 300 and 350 nm. For this range, the wavelength average quantum yield for BDE209 photoreaction, phiave, was calculated to be 0.47. The difference between kobs values at the two exposure times is explained fully by the difference between the solar irradiation fluxes. Upon solar irradiation, BDE209 reductively dehalogenated to other polybrominated diphenyl ethers (PBDEs). During 34 h of irradiation, PBDEs ranging from nona- to tri-bromodiphenyl ethers were observed. In total, 43 PBDEs were detected, and the GC retention times and mass spectral fragment patterns of 21 products matched those of available congener standards, including congeners 2,2',4,4',5-pentabromodiphenyl ether and 2,2',4,4' tetrabromodiphenyl ether. PMID- 15352455 TI - Secondary organic aerosol formation from the ozonolysis of cycloalkenes and related compounds. AB - The secondary organic aerosol (SOA) yields from the laboratory chamber ozonolysis of a series of cycloalkenes and related compounds are reported. The aim of this work is to investigate the effect of the structure of the hydrocarbon parent molecule on SOA formation for a homologous set of compounds. Aspects of the compound structures that are varied include the number of carbon atoms present in the cycloalkene ring (C5 to C8), the presence and location of methyl groups, and the presence of an exocyclic or endocyclic double bond. The specific compounds considered here are cyclopentene, cyclohexene, cycloheptene, cyclooctene, 1 methyl-1-cyclopentene, 1-methyl-1-cyclohexene, 1-methyl-1-cycloheptene, 3-methyl 1-cyclohexene, and methylenecyclohexane. The SOA yield is found to be a function of the number of carbons present in the cycloalkene ring, with an increasing number resulting in increased yield. The yield is enhanced by the presence of a methyl group located at a double-bonded site but reduced by the presence of a methyl group at a non-double-bonded site. The presence of an exocyclic double bond also leads to a reduced yield relative to that of the equivalent methylated cycloalkene. On the basis of these observations, the SOA yield for terpinolene relative to the other cyclic alkenes is qualitatively predicted, and this prediction compares well to measurements of the SOA yield from the ozonolysis of terpinolene. This work shows that relative SOA yields from ozonolysis of cyclic alkenes can be qualitatively predicted from properties of the parent hydrocarbons. PMID- 15352456 TI - Chlorophyll fluorescence imaging of individual algal cells: effects of herbicide on Spirogyra distenta at different growth stages. AB - Serious environmental degradation of aquatic ecosystems has been caused by eutrophication and by pollutants such as herbicides. Therefore, measurement of in situ algal photosynthetic activity is important for environmental monitoring. With ordinary nonimaging fluorometers, algal chlorophyll fluorescence can be measured easily, but heterogeneity within samples cannot be detected. Effects of a herbicide preparation containing 3-(3,4-dichlorophenyl)-1,1 -dimethylurea (DCMU) on photosynthetic activity at different growth stages of Spirogyra distenta were investigated by using a computer-aided microscopic imaging system for chlorophyll afluorescence. Photosystem II photochemical yield (phiPSII) images were used to diagnose photosynthetic activity of spiral filate chloroplasts in algal cells. The herbicide treatment caused a stronger decline in phiPSII values in younger than in mature algae cells. This result indicated that heterogeneity within algal samples should be considered when algae are used for environmental monitoring. Thus, measurement of chlorophyll fluorescence from young and mature chloroplasts with a microscopic imaging system makes it possible to improve the sensitivity for monitoring the environmental degradation of aquatic ecosystems. PMID- 15352457 TI - Desorption kinetics of phenanthrene in aquifer material lacks hysteresis. AB - Desorption experiments were carried out in flow through columns following long term sorption batch experiments (up to 1010 days at 20 degrees C; Rugner, H.; Kleineidam, S.; Grathwohl, P. Long-term sorption kinetics of phenanthrene in aquifer materials. Environ. Sci. Technol. 1999, 33, 1645-1651) to elucidate sorption/desorption hysteresis phenomena of phenanthrene in aquifer materials. Most of the sorbents employed in this study (homogeneous lithocomponents separated from aquifer sediments or fresh rock fragments) showed highly nonlinear sorption isotherms because of coal particles embedded inside the grains. Because sorption capacities were high, sorption equilibrium was not reached in most of the sorbents during the initial sorptive uptake experiments lasting up to 1010 days. Desorption was studied up to 90 days at 20 degrees C. The temperature was raised after that stepwise from originally 20 to 30, 40, 50, and finally to 70 degrees C for selected samples to estimate activation energies of desorption. A numerical intraparticle pore diffusion model was used to fit sorptive uptake data and subsequently for pure forward prediction of the release rates in the desorption column experiments. Desorption was initially fast followed by extended tailing which in other studies is fitted by using multirate first-order models. Our results demonstrate that the retarded intraparticle pore diffusion model can predict the desorption rates with a single diffusion rate constant obtained independently from the long-term batch sorption experiment. No evidence for hysteresis was found, suggesting that many hysteresis phenomena reported earlier are experimental artifacts resulting from nonequilibrium effects and "nonphysical" models. The different temperature steps allowed one to additionally calculate activation energies of desorption (45-59 kJ mol(-1)), which were in reasonably good agreement with results from earlier studies for a retarded pore diffusion process. In addition, equilibrium sorption isotherms were determined at 20 and 40 degrees C to compare sorption and desorption enthalpies. Both were in good agreement, confirming that desorption was not significantly different from sorption. PMID- 15352458 TI - Simulating the influence of snow on the fate of organic compounds. AB - Snow scavenging, a seasonal snowpack, and a dynamic water balance are incorporated in a non-steady-state generic multimedia fate model in order to investigate the effect of snow on the magnitude and temporal variability of organic contaminant concentrations in various environmental media. Efficient scavenging of large nonpolar organic vapors and particle-bound organic chemicals by snow can lead to reduced wintertime air concentrations and incorporation in the snowpack. The snow cover functions as a temporary storage reservoir that releases contaminants accumulating over the winter during a short melt period, resulting in temporarily elevated concentrations in air, water, and soil. The intensity of these peaks increases with the length of the snow accumulation period. Organic chemicals of sufficient volatility (log KOA < 9; e.g., light polychlorinated biphenyls) can volatilize from the snowpack, resulting in springtime concentration maxima in the atmosphere. The behavior of fairly water soluble chemicals during snowmelt depends on their relative affinity for the newly formed liquid water phase and the rapidly diminishing ice surface quantitatively expressed by their interface-water partition coefficient (KIW). Chemicals with a preference for the dissolved phase (low KIW; e.g., pentachlorophenol) can become enriched in the first meltwater fractions and experience a temporary concentration peak in lakes and rivers. Organic chemicals that are neither volatile enough to evaporate from the snowpack nor sufficiently water soluble to dissolve in the meltwater (e.g., polybrominated diphenyl ethers) sorb to the particles in the snowpack. These particles may be sufficiently contaminated to constitute the major input route to the terrestrial environment upon release during snowmelt. Because wintertime deposition to the snowpack may be higher than to a non-snow covered surface, this can result in higher soil concentrations of persistent organic contaminants in the long term. The potential ecotoxicological significance of peak exposures demands a better understanding of the role of snow in the fate of organic contaminants. PMID- 15352459 TI - Sources of toxaphene and other organochlorine pesticides in North America as determined by air measurements and potential source contribution function analyses. AB - A previous study from our laboratory suggested that the Gulf of Mexico might be a potential source of toxaphene to the United States. To investigate this hypothesis, we measured gas-phase toxaphene concentrations at sampling sites ranging from northern Michigan to southern Louisiana; the samples were collected every 12 days during 2002-2003. We also measured other organochlorine pesticides in these samples. We identified major source regions of each pesticide group using the potential source contribution function model with the Clausius Clapeyron equation defining the criterion levels. These results indicate that southern cotton farms are major sources of both toxaphene and p,p'-DDE to the northern United States. In fact, there is a very strong correlation of toxaphene and DDE atmospheric concentrations at all sites, further indicating a common source. On the other hand, the Gulf of Mexico is not a major source of toxaphene or DDE. DDE's source region is similar to that of toxaphene but somewhat broader, reflecting DDT's historically more diversified use. The level of endosulfan in the atmosphere at all of the sites was similar, and PSCF modeling indicated that its sources were all toward the east of the sampling sites. PMID- 15352460 TI - A novel analytical approach for visualizing and tracking organic chemicals in plants. AB - Vegetation plays a key role in the environmental fate of many organic chemicals, from pesticides applied to plants, to the air-vegetation exchange and global cycling of atmospheric organic contaminants. Our ability to locate such compounds in plants has traditionally relied on inferences being made from destructive chemical extraction techniques or methods with potential artifacts. Here, for the first time, two-photon excitation microscopy (TPEM) is coupled with plant autofluorescence to visualize and track trace levels of an organic contaminant in living plant tissue, without any form of sample modification or manipulation. Anthracene-a polynuclear aromatic hydrocarbon (PAH)-was selected for study in living maize (Zea mays) leaves. Anthracene was tracked over 96 h, where amounts as low as approximately 0.1-10 pg were visible, as it moved through the epicuticular wax and plant cuticle, and was observed reaching the cytoplasm of the epidermal cells. By this stage, anthracene was identifiable in five separate locations within the leaf: (1) as a thin (approximately 5 microm) diffuse layer, in the upper surface of the epicuticular wax; (2) as thick (approximately 28 microm) diffuse bands extending from the epicuticular wax through the cuticle, to the cell walls of the epidermal cells; (3) on the external surface of epidermal cell walls; (4) on the internal surface of epidermal cell walls; and (5) within the cytoplasm of the epidermal cells. This technique provides a powerful nonintrusive tool for visualizing and tracking the movement, storage locations, and degradation of organic chemicals within vegetation using only plant and compound autofluorescence. Many other applications are envisaged for TPEM, in visualizing organic chemicals within different matrixes. PMID- 15352461 TI - Development and validation of a canister method for measuring ethylene oxide in ambient air. AB - A sampling and analytical method for measuring ethylene oxide (EO) in ambient air was developed and evaluated. The method is based on the use of evacuated canisters and gas chromatography-mass spectrometry (GC-MS). The objectives of this work were to characterize the performance of the method with respect to the following: (1) stability/recovery of ethylene oxide in a canister over a 15-day holding time; (2) detection capability; and (3) measurement of EO in an ambient air matrix. Both electropolished and silica-lined stainless steel canisters were evaluated in this study. The method evaluation involved both laboratory and field tests. The recovery of the EO was evaluated both on an absolute basis and relative to a spiked internal standard of toluene. EO spiked at levels of 2 ppbv and 20 ppbv was found to be stable for holding times of up to 15 days at 25 degrees C in both a humidified nitrogen matrix and in ambient air. The detection limit of the method was found to be 0.25 ppbv using EPA's traditional approach of seven replicate analyses of a low-level standard and 0.20 ppbv using a probability-based approach. EO recoveries in the laboratory stability study generally were 100 +/- 25%, and did not vary by canister type, nor did the EO recoveries decrease with holding time. Field studies demonstrated that the method is capable of detecting EO (as well as benzene and toluene) in an ambient air matrix. PMID- 15352462 TI - Tire-wear particles as a source of zinc to the environment. AB - Tire-tread material has a zinc (Zn) content of about 1 wt %. The quantity of tread material lost to road surfaces by abrasion has not been well characterized. Two approaches were used to assess the magnitude of this nonpoint source of Zn in the U.S. for the period 1936-1999. In the first approach, tread-wear rates from the automotive engineering literature were used in conjunction with vehicle distance-driven data from the U.S. Department of Transportation to determine Zn releases. A second approach calculated this source term from the volume of tread lost during lifetime tire wear. These analyses showed that the quantity of Zn released by tire wear in the mid-1990s was of the same magnitude as that released from waste incineration. For 1999, the quantity of Zn released by tire wear in the U.S. is estimated to be 10 000-11 000 metric tons. A specific case study focused on Zn sources and sinks in an urban-suburban watershed (Lake Anne) in the Washington, DC, metropolitan area for a time period of the late 1990s. The atmospheric flux of total Zn (wet deposition) to the watershed was 2 microg/cm2/yr. The flux of Zn to the watershed estimated from tire wear was 42 microg/cm2/yr. The measured accumulation rate of total Zn in age-dated sediment cores from Lake Anne was 27 microg/cm2/yr. These data suggest that tire-wear Zn inputs to urban-suburban watersheds can be significantly greater than atmospheric inputs, although the watershed appears to retain appreciable quantities of vehicular Zn inputs. PMID- 15352463 TI - Retention of three heavy metals (Zn, Pb, and Cd) in a calcareous soil controlled by the modification of flow with geotextiles. AB - Although geotextiles are increasingly employed in stormwater infiltration basins, their influence on the flow and transfer of contaminants, such as heavy metals, has not been fully investigated. Leaching column experiments were conducted to characterize the flow and transfer of three heavy metals (zinc, lead, and cadmium) in a calcareous soil with and without geotextiles under steady-state flow and close to saturation forthe soil. The influence of geotextiles was characterized for two types of geotextiles (needlepunched and thermosealed) and for two different initial saturation degrees for the needlepunched geotextile. The main results showed that, when placed wet, the needlepunched geotextile had no influence. When placed dry, it homogenized the flow in its surroundings and thus allowed better contact between heavy metals and the reactive soil, resulting in an increase of their retention. The thermosealed geotextile, placed dry, homogenized the flow and increased retention over a larger area, resulting in optimal global retention. In conclusion, geotextiles could be used in infiltration basins, provided that their effect on both flow and heavy metal retention is optimized by appropriate design--choice of geotextiles--and appropriate monitoring--control of hydric conditions. PMID- 15352464 TI - Formation of PCDD/Fs in the sintering process: influence of the raw materials. AB - The sintering process is among the major sources of PCDD/Fs in the environment. This research studies the influence of the raw materials in this type of industrial plant on the amounts of PCDD/Fs generated. Particular interest is given to coke, which constitutes the principal source of carbon for the de novo synthesis of PCDD/Fs, and to the dust collected in the electrostatic precipitator (E.S.P. dust), usually recycled in the raw materials. The de novo synthesis of PCDD/Fs is simulated at the laboratory scale by thermal treatments of the samples. The use of a particular coke as a fuel does not drastically reduce the formation of PCDD/Fs. Actually, the global amounts of PCDD/Fs generated from the graphite and the two cokes tested are very similar. Only modifications in the fingerprint are observed. On the other hand, the addition of 10 wt % dust collected in the electrostatic precipitator leads to the formation of amounts of PCDD/Fs multiplied by a factor larger than 10(3). These results imply caution against the recycling of this E.S.P. dust in the raw materials. PMID- 15352465 TI - Evaluation of current techniques for isolation of chars as natural adsorbents. AB - Chars in soils or sediments may potentially influence the soil/sediment sorption behavior. Current techniques for the isolation of black carbon including chars rely often on acid demineralization, base extraction, and chemical oxidation to remove salts and minerals, humic acid, and refractory kerogen, respectively. Little is known about the potential effects of these chemical processes on the char surface and adsorptive properties. This study examined the effects of acid demineralization, base extraction, and acidic Cr2O7 2- oxidation on the surface areas, surface acidity, and benzene adsorption characteristics of laboratory produced pinewood and wheat-residue chars, pure or mixed with soils, and a commercial activated carbon. Demineralization resulted in a small reduction in the char surface area, whereas base extraction showed no obvious effect. Neither demineralization nor base extraction caused an appreciable variation in benzene adsorption and presumably the char surface properties. By contrast, the Cr2O7 2- oxidation caused a >31% reduction in char surface area. The Boehm titration, supplemented by FTIR spectra, indicated that the surface acidity of oxidized chars increased by a factor between 2.3 and 12 compared to non-oxidized chars. Benzene adsorption with the oxidized chars was lower than that with the non oxidized chars by a factor of >8.9; both the decrease in char surface area and the increase in char surface acidity contributed to the reduction in char adsorptive power. Although the Cr2O7 2-oxidation effectively removes resistant kerogen, it is not well suited for the isolation of chars as contaminant adsorbents because of its destructive nature. Alternative nondestructive techniques that preserve the char surface properties and effectively remove kerogen must be sought. PMID- 15352466 TI - Chitosan-supported palladium catalyst. 5. Nitrophenol degradation using palladium supported on hollow chitosan fibers. AB - Hollow chitosan fibers were reacted with chloropalladate solutions and subsequently reduced by hydrogen produced in situ by reaction of sulfuric acid with zinc powder in order to manufacture palladium supported on catalytic hollow chitosan fibers (C2HF-Pd). This catalytic support was used to degrade 3 nitrophenol (3-NP) using two different hydrogen donors (hydrogen gas and sodium formate). The solution was flowed through the lumen of the fiber, while the sodium formate was recirculated round the outside of the fiber. In the case of hydrogen gas, the gas was maintained under controlled pressure outside the fiber. The influence of the pH, residence time (ca. flow velocity), nitrophenol concentration, and hydrogen-donor concentration (or pressure) was investigated for both systems in order to evaluate the limiting parameters. While the system using sodium formate was the most efficient for nitrophenol conversion, the system using hydrogen gas avoided the production of secondary waste solutions (formate solutions with traces of nitrophenol, which pass through the fiber membrane). PMID- 15352467 TI - Degradation and sorption of selected organophosphate and carbamate insecticides in urban stream sediments. AB - Monitoring studies show that urban surface streams in the United States are commonly contaminated with pesticides, and contamination by organophosphates and carbamates is of particular concern because of their aquatic toxicity. The degradation and sorption of four common organophosphate and carbamate insecticides were studied in urban creek sediments from southern California, USA. In sediment, malathion was quickly degraded under either aerobic or anaerobic conditions, with a half-life (t(1/2)) <3 d. Diazinon and chlorpyrifos were moderately persistent under aerobic conditions (t(1/2) = 14-24 d). However, persistence of chlorpyrifos increased significantly under anaerobic conditions, and t(1/2) was prolonged to 58 to 223 d. The greatest effect of redox potential was found with carbaryl. Although rapid dissipation occurred under aerobic conditions (t(1/2) = 1.8-4.9 d), carbaryl became virtually nondegradable under anaerobic conditions (t(1/2) = 125-746 d). The sorption coefficient consistently increased with time for all pesticides, and chlorpyrifos displayed greater sorption potential than the other pesticides. This study indicates that pesticides in sediment may become less available with time because of increased sorption, and pesticide persistence in sediment may vary greatly among compounds and with redox conditions. Under anaerobic conditions, long persistence may occur even for nonpersistent compounds. PMID- 15352468 TI - Organophosphorus pesticides in storm-water runoff from southern California (USA). AB - Large quantities of the organophosphorus (OP) pesticides diazinon and chlorpyrifos are applied to California (USA) watersheds every year, but few data are available on the sources of OP pesticides in urban watersheds. The goal of this study was to characterize diazinon and chlorpyrifos concentrations from different land uses indicative of source categories in urban southern California watersheds. This characterization included analysis of 128 runoff samples from eight different land uses over five storm events. Diazinon was consistently detected (93% of samples) during this study, whereas chlorpyrifos was not consistently detected (12% of samples). The mixed agricultural land use had the highest flow weighted mean (FWM) concentration of diazinon (4076 ng/L), which exceeded the next-highest land-use categories (commercial and residential) by one to two orders of magnitude (324-99 ng/L, respectively). Open space had the lowest concentration of diazinon (<20 ng/L). Concentrations of diazinon at replicate land-use sites and during replicate storm events at the same site were highly variable. The difference in diazinon FWM concentrations among replicate sites ranged from 1.5-fold to 45-fold. The difference in diazinon FWM concentrations among storms at the same site ranged from 1.25-fold to 30-fold. Part of this variability is a response to the temporal patterns observed within a storm event. The majority of land-use site-events had peak concentrations before peak flow indicating a first-flush effect, but this was not always a predictable temporal trend. The first-flush effect was rarely evident in terms of mass loadings because flows can range orders of magnitude during a single event in highly impervious urban watersheds. Flow variability thus overwhelms the variability in diazinon concentrations attributable to the first-flush effect. PMID- 15352469 TI - Water-extractability, free ion activity, and pH explain cadmium sorption and toxicity to Folsomia candida (Collembola) in seven soil-pH combinations. AB - Toxicity of cadmium to Folsomia candida was determined in soils at different pHs (3.5, 5.0, and 6.5). The Langmuir sorption constant (K(L)), based on pore-water or water-extractable concentrations, showed a pH-related increase of cadmium sorption that was most pronounced when using free Cd2+ ion activities ([Cd2+]s). Two-species Langmuir isotherms that used total cadmium concentration ([Cd]) or [Cd2+] and pH in the water-extractable fractions gave the best description of cadmium sorption on all soils together. Cadmium concentrations causing 50% reduction of growth and reproduction (median effective concentrations [EC50s]) differed by a factor of 4.5 to 20 when based on total soil concentrations and increased with increasing pH. However, when based on water-extractable or pore water [Cd] or [Cd2+], EC50s decreased with increasing pH, but differences between soils were still a factor of 4.5 to 32. The EC50s differed by less than a factor of 2.2 when based on body [Cd] in the surviving animals. Two-species Langmuir isotherms were used to relate body [Cd] in survivors to [Cd2+], corrected for pH in water-extractable or pore-water fractions. An excellent description of effects on growth and reproduction was found when related to the body concentrations predicted in this way; the difference in EC50s between soils was reduced to a factor <2. This demonstrates that F. candida is mainly exposed to cadmium through the soil solution, and suggests that principles of a biotic ligand model approach may be applicable for this soil organism. PMID- 15352470 TI - Effect of extraction conditions on deposit feeder in trace element solubilization in only a digestive fluid. AB - In vitro extraction of sediments using digestive fluid of deposit-feeding invertebrates has been advocated as a measure of in vivo bioavailability, but the standard procedure used for the extractions alters fluid properties relative to in vivo conditions. In vivo fluid properties (redox potential [Eh], pH, dissolved oxygen) were measured in the gut of the polychaete Arenicola brasiliensis, and then the effects of gut evacuation, elevated temperature, and atmospheric exposure on these properties and/or metal and metalloid extraction capability of the fluid was assessed.The midgut of A. brasiliensis has an electropositive Eh of about +170 to +230 mV, a neutral pH, and little or no dissolved oxygen. Exposure to the atmosphere during in vitro extraction increased gut fluid oxygen content and affected the ability of the fluid to extract some trace elements from sediment particles, particularly Pb, Ni, Cu, Mn, and As. Even for these substances, however, the effects of oxygen exposure on extractability by gut fluid typically were small (<2-fold difference in amount extracted). For Cr, Hg, Se, Cd, Fe, and Zn, the effects of oxygen exposure usually were minor and inconsistent in direction. Maintaining anaerobic conditions during in vitro contaminant extractions is necessary when the experimental objective is to mimic accurately in vivo conditions and predict trace element extraction in the gut. If using metal extractability as a relative measure for sediment risk assessment purposes, a high degree of accuracy may not be required, and aerobic in vitro extractions could be suitable and produce only minor errors in extractability relative to in vivo conditions. PMID- 15352471 TI - The role of desorption for describing the bioavailability of select polycyclic aromatic hydrocarbon and polychlorinated biphenyl congeners for seven laboratory spiked sediments. AB - Lumbriculus variegatus and Diporeia spp. were exposed to two contaminant pairs 3H benzo[a]pyrene (BaP) and 14C-2,4,5,2'4',5'-hexachlorobiphenyl (HCBP), and 3H pyrene (PY) and 14C-3,4,3',4'-tetrachlorobiphenyl (TCBP) sorbed to each of seven field-collected sediments of varying composition. Toxicokinetic coefficients, bioaccumulation factors (BAF), and biota-sediment accumulation factors ([BSAF], BAF normalized to the organism lipid content and sediment organic carbon content) were determined. The contaminant desorption rates from sediments were measured with a Tenax resin extraction technique. The desorption rate constants for rapid, slow, and very slow contaminant compartments and the fractions of contaminant in each compartment were compared with the toxicokinetic coefficients, BAF, and BSAF to describe contaminant behavior across sediments, among compounds, and between species. The best description of the bioavailability was the log BSAF regressed against the fraction rapidly desorbed (F(rapid)) across all sediments and compounds for both species, r2 = 0.67 and 0.66 for L. variegatus and Diporeia, respectively. Use of a calculated fraction desorbed in 6 h or 24 h, which has been suggested as a surrogate for F(rapid,) did not produce as predictive a regression because of uneven desorption in a fixed duration for each compound among the sediments. Thus, F(rapid)provided a good surrogate for the bioavailability of the sediment-sorbed contaminant as represented by BSAF across seven sediments and four compounds with predictions within a factor of approximately two of the measured value. PMID- 15352472 TI - Fate and metabolism of [15N]2,4,6-trinitrotoluene in soil. AB - The fates of the labels from [14C] and [15N] trinitrotoluene were analyzed in bioreactors under aerobic conditions in soil treated by a fungal bioremediation process with Stropharia rugosoannulata and in control soil. Up to 17.5% of the 15N label had a different fate than the 14C label. Three N-mineralization processes were identified in detailed experiments with [15N]TNT. About 2% of the 15N label was found as NO3- and NH4+, showing simultaneous processes of direct TNT denitration (I) and reduction with cleavage of the amino groups (II). The enrichment of NO2-/NO3- (up to 7.5 atom% 15N abundance) indicates the formation of Meisenheimer complexes with a denitration of [15N]TNT. A 1.4% of the label was found distributed between N2O and N2. However, the 15N enrichment of the N2O (up to 38 atom%) demonstrated that both N atoms were generated from the labeled TNT and clearly indicates a novel formation process (III). We propose, as an explanation, the generation of N2O by cleavage from condensed azoxy metabolites. In addition, 1.7% of the 15N label was detected as biogenic amino acids in the wheat straw containing the fungus. Overall, 60 to 85% of the applied [15N]TNT was degraded and 52 to 64% was found as nonextractable residues in the soil matrix. Three percent was detected as 2-amino-4,6-dinitrotoluene and 4-amino-2,6 dinitrotoluene. PMID- 15352473 TI - Assessment of the availability of polycyclic aromatic hydrocarbons from gasworks soil using different extraction solvents and techniques. AB - This study was designed to assess the availability of polycyclic aromatic hydrocarbons (PAHs) present at a gasworks site to different soil remediation techniques. The study examined the effect on PAH availability of using different organic solvents, the degree of pretreatment, and the extraction time. In total, 25 PAHs (with two to six fused rings) and five carbonyl derivatives were measured. The results indicated that the PAHs and their derivatives were bound loosely to the surface of the studied soil and that there were no significant kinetic boundaries associated with the extraction of the PAHs. Furthermore, it was concluded that the studied soil was not suitable for bioremediation, as the concentration of PAHs with low molecular weight were limited. However, pressurized liquid extraction (PLE) with methanol as the solvent extracted 97% of all PAHs and PAH-derivatives, indicating that extraction may be effective as part of a soil remediation technique for old gasworks soils. PMID- 15352474 TI - Blarina brevicauda as a biological monitor of polychlorinated biphenyls: evaluation of hepatic cytochrome P450 induction. AB - We assessed the value of short-tailed shrews (Blarina brevicauda) as a possible biomonitor for polychlorinated biphenyl pollution through measurement of the induction of hepatic cytochrome P450 and associated enzyme activities. First, we checked the inducibility of four monooxygenases (benzyloxyresorufin-O-dealkylase [BROD], ethoxyresorufin-O-dealkylase [EROD], methoxyresorufin-O-dealkylase [MROD], and pentoxyresorufin-O-dealkylase [PROD]) by measuring the activity of these enzymes in hepatic microsomes prepared from shrews injected with beta naphthoflavone (betaNF) or phenobarbital (PB), typical inducers of cytochrome P4501A (CYP1A) and CYP2B enzyme families, respectively. Enzyme activity was induced in shrews that received betaNF but not in shrews that received PB; PROD was not induced by either exposure. Later, shrews were exposed to a mixture of polychlorinated biphenyls (PCBs) (Aroclor 1242:1254, in 1:2 ratio) at 0.6, 9.6, and 150 ppm in food, for 31 d. Induction in these shrews was measured by specific enzyme activity (BROD, EROD, and MROD) in hepatic microsomes, by western blotting of solubilized microsomes against antibodies to CYP1A or CYP2B, and by duration of sodium pentobarbital-induced sleep. These three CYP enzymes were induced in shrews by PCBs at similar levels of exposure as in cotton rat (Sigmodon hispidus). Neither sleep time nor the amount of CYP2B family protein were affected by PCB exposure. Blarina brevicauda can be a useful biomonitor of PCBs that induce CYP1A, especially in habitats where they are the abundant small mammal. PMID- 15352475 TI - Estrogenic potential of the Venice, Italy, lagoon waters. AB - The exposure of the Venice lagoon (Italy) to endocrine-disrupting compounds (EDCs) from different sources was investigated. Spatial and time distribution of EDC concentrations were determined in four sampling sessions (December 2001-May 2002) by solid phase extraction followed by high-performance liquid chromatography separation coupled with mass spectrometry detection via electrospray interface (SPE-HPLC-ESI-MS), which allowed identification of natural (estradiol, estrone) and synthetic estrogenic compounds, both steroidal (ethinylestradiol, mestranol) and nonsteroidal (benzophenone, bisphenol-A, nonylphenol, nonylphenol monoethoxylate carboxylate). No significant differences in the EDC distribution were observed between stations located near selected sources (raw sewage from the historical center of Venice, treated municipal and industrial effluents from sewage treatment plants, and areas undergoing the inflow of rivers). While synthetic nonsteroidal analytes were recorded in the 1 to 1040 ng/L range (average concentration: 34 ng/L), steroidal EDC (estradiol, ethinylestradiol) concentrations were lower (1-125 ng/L; average concentration: 8 ng/L). The estrogenic activity of lagoon waters was estimated in terms of estradiol equivalent concentration (EEQ) by applying the estradiol equivalency factors (EEFs). Steroidal EDCs (estradiol, ethinylestradiol) contributed >97% to the total potential estrogenicity of the waters, which accounted for 4 to 172 ng/L (average: 25 ng/L), as total EEQs. These levels are likely to pose adverse effects on the Venice lagoon aquatic organisms. PMID- 15352476 TI - Arsenic speciation in tissues of the Mediterranean polychaete Sabella spallanzanii. AB - Arsenic toxicity is strictly related to its chemical form and marine organisms are known to accumulate this element mostly as organoarsenic nontoxic molecules. Contrasting with this general trend, the presence of moderately toxic to toxic arsenic compounds recently has been reported in some polychaete species, showing a completely different profile of represented chemical species. In this work the presence and distribution of arsenic were characterized in the Mediterranean polychaete Sabella spallanzanii, by analyzing total levels in different tissues and subcellular fractions and the occurrence of various arsenical compounds. Further investigations on arsenic accumulation in S. spallanzanii were based on the capability of this species to regenerate the branchial crown both in laboratory and field conditions. Though basal levels of arsenic in the thorax were similar to those already described for most polychaetes and invertebrate species, branchial crown revealed a remarkable accumulation of this element with concentrations higher than 1000 microg/g. Arsenic mainly was localized in a soluble form within the cytosol and dimethyl-arsinate (DMA) appeared the most represented chemical species. Experiments on bioaccumulation of arsenic in regenerating branchial crowns confirmed the environmental origin of this element and the time-dependent appearance of various As compounds suggested a methylation pathway more than a degradation process for the elevated content of DMA. The accumulation of this moderately toxic compound in the more vulnerable tissues of the polychaete might represent an antipredatory strategy, as indicated by some feeding trials where fish ate the thorax but rejected the branchial crowns. PMID- 15352477 TI - Use of cyclodextrin and calcium chloride for enhanced removal of mercury from soil. AB - The use of solutions containing carboxymethyl-beta-cyclodextrin (CMCD) or CaCl2 for enhancing the removal of Hg from a sandy soil was investigated using batch and column experiments. The retention of Hg appeared to be controlled by specific adsorption reactions, which greatly constrained Hg removal when using water (KNO3 solution) to flush columns packed with contaminated soil. The results showed that the two reagents did enhance the removal of Hg from the soil. For example, 81% and 60% of Hg was recovered after 50 pore volumes of flushing with 50 mM CaCl2 and 2 mM CMCD, respectively, compared to 24% recovery for a 10 mM KNO3 solution. However, significant tailing and delayed recovery of Hg during the elution process occurred in the presence of all reagents, indicating that the removal of Hg from the soil was rate limited. PMID- 15352478 TI - Degradation of petroleum hydrocarbons in two sub-antarctic soils: influence of an oleophilic fertilizer. AB - In order to determine the long-term effects of fertilizer on the degradation rate and the toxicity of hydrocarbons in sub-Antarctic soils contaminated by petroleum hydrocarbons, a field study was initiated in December 2000 on two different soils of the Kerguelen Islands (69 degrees 42'E, 49 degrees 19'S). The number of hydrocarbon-degrading bacteria (HDB) increased greatly after crude-oil and diesel fuel contamination, and the fertilizer addition had a favorable effect on HDB growth and activity. Hydrocarbon-degrading bacteria counts remained high until the end of the experiment although the total hydrocarbon content in all contaminated soils was reduced to 80 to 90% of their initial value after 330 d. Degradation of n-alkanes was enhanced significantly in the presence of the fertilizer, while the degradation of polycyclic aromatic hydrocarbons (PAHs) was only barely enhanced. Toxicity results showed a noticeable reduction with time, although toxicity remained present and important in both soils at the end of the experiment. In addition, fertilized plots showed a toxic signal greater than unfertilized ones. Overall results clearly show that fertilizer addition improves the rate of degradation of both oil contaminants. However, remaining toxic residues may constitute a drawback of the fertilizer-assisted biodegradation process at low temperatures. PMID- 15352479 TI - Reduced growth of rainbow trout (Oncorhynchus mykiss) fed a live invertebrate diet pre-exposed to metal-contaminated sediments. AB - Juvenile rainbow trout (Oncorhynchus mykiss) were fed live diets of Lumbriculus variegatus cultured in metal-contaminated sediments from the Clark Fork River Basin (MT, USA), an uncontaminated reference sediment, or an uncontaminated culture medium. Fish were tested in individual chambers; individual growth as well as the nutritional quality and caloric value of each trout's consumed diet were determined. Growth was measured following 14, 28, 42, 56, and 67 d of exposure. A subset of fish was sampled at 35 d for whole-body metals. Metals (whole body, digestive tract, and liver) and histology were measured at the end of the test. We observed significant growth inhibition in trout fed the contaminated diets; growth inhibition was associated with reductions in conversion of food energy to biomass rather than with reduced food intake. Growth inhibition was negatively correlated with As in trout tissue residues. Histological changes in contaminated treatments included hepatic necrosis and degenerative alterations in gallbladder. The present study provides evidence that metal-contaminated sediments can pose a hazard to trout health through a dietary exposure pathway. PMID- 15352480 TI - Impact of perfluorooctanoic acid on fathead minnow (Pimephales promelas) fatty acyl-CoA oxidase activity, circulating steroids, and reproduction in outdoor microcosms. AB - This study investigates reproductive impairment and biochemical changes in fathead minnow (Pimephales promelas) exposed for 39 d to varying concentrations of perfluorooctanoic acid (PFOA) under microcosm conditions. While the concentrations tested in this study were much higher than those normally found in the environment, no mortality was associated with PFOA exposure. Only modest changes were observed in condition factor and in relative liver and gonad size. Significant declines in circulating plasma steroids were observed, but these were accompanied by only limited increases in time to first oviposition and decreases in overall egg production. Peroxisome proliferation, as quantified by fatty acyl CoA oxidase (FAO) activity, was elevated with low PFOA concentrations but attenuated with exposure to higher PFOA doses. Little evidence was seen of differential induction of peroxisome-associated enzyme activity with sex. Oxidative stress, as quantified by the 2-thiobarbituric acid reactive substances (TBARS) assay, was only modestly influenced by PFOA exposure and is not a significant consequence of FAO activity in fathead minnow. Perfluorooctanoic acid appears to be relatively nontoxic at environmentally relevant concentrations but may impact biochemical and reproductive endpoints under conditions associated with environmental spills. PMID- 15352481 TI - Effects of the pesticides captan, deltamethrin, isoproturon, and pirimicarb on the microbial community of a freshwater sediment. AB - In three microcosm experiments, we exposed microbial communities of a natural sediment to environmentally relevant concentrations of the fungicide captan, the herbicide isoproturon, and the insecticides deltamethrin and pirimicarb. Exposure concentrations were estimated negligible concentrations (NCs), maximum permissible concentrations (MPCs), and 100 times MPC (100MPC). Experimental endpoints were microbial community respiration and biomass, bacterial activity, and denitrification. All four pesticides inhibited bacterial activity by 20 to 24% at MPC, which corresponded to concentrations in the range of microg/kg dry weight sediment. Treatments with deltamethrin and isoproturon showed inhibiting effects on bacterial activity at NC exposures. Surprisingly, for captan, deltamethrin, and isoproturon, this inhibiting effect was not observed at 100MPC treatments. Microbial biomass was negatively effected in MPC treatments with deltamethrin and in NC treatments with isoproturon. The tested pesticides did not affect community respiration and denitrification rates. These results show that exposure to the tested pesticides may induce toxic responses in sediment microbial communities at concentrations that are predicted to be environmentally safe. PMID- 15352482 TI - Toxicity of glyphosate-based pesticides to four North American frog species. AB - Glyphosate-based herbicides are among the most widely used pesticides in the world. We compared the acute toxicity of the glyphosate end-use formulation Roundup Original to four North American amphibian species (Rana clamitans, R. pipiens, R. sylvatica, and Bufo americanus) and the toxicity of glyphosate technical, the polyethoxylated tallowamine surfactant (POEA) commonly used in glyphosate-based herbicides, and five newer glyphosate formulations to R. clamitans. For R. clamitans, acute toxicity values in order of decreasing toxicity were POEA > Roundup Original > Roundup Transorb > Glyfos AU; no significant acute toxicity was observed with glyphosate technical material or the glyphosate formulations Roundup Biactive, Touchdown, or Glyfos BIO. Comparisons between the four amphibian species showed that the toxicity of Roundup Original varied with species and developmental stage. Rana pipiens tadpoles chronically exposed to environmentally relevant concentrations of POEA or glyphosate formulations containing POEA showed decreased snout-vent length at metamorphosis and increased time to metamorphosis, tail damage, and gonadal abnormalities. These effects may be caused, in some part, by disruption of hormone signaling, because thyroid hormone receptor beta mRNA transcript levels were elevated by exposure to formulations containing glyphosate and POEA. Taken together, the data suggest that surfactant composition must be considered in the evaluation of toxicity of glyphosate-based herbicides. PMID- 15352483 TI - Dietary accumulation and metabolism of polybrominated diphenyl ethers by juvenile carp (Cyprinus carpio). AB - Polybrominated diphenyl ethers (PBDEs) are hydrophobic organic contaminants with properties and nomenclature similar to polychlorinated biphenyls (PCBs). While much information is available on the bioaccumulation and pharamcokinetics of PCBs, little information is available on PBDEs. In this study, juvenile carp were exposed to a diet spiked with a cocktail of four BDE congeners (2,4,4'-tribromoDE [BDE 28], 2,2',4,4'-tetrabromoDE [BDE 47], 2,2',4,4',5-pentabromoDE [BDE 99], and 2,2',4,4',5,5'hexabromoDE [BDE 153]) for 60 d followed by a 40-d depuration period. As a positive control, three PCB congeners with similar log K(ow), values (2,2',5,5'-tetrachlorobiphenyl [PCB 52], 2,2',4,4', 5,5'-hexachlorobiphenyl [PCB 153], and 2,2',3,4,4',5,5'-heptachlorobiphenyl [PCB 180]) were included in the cocktail to compare their assimilation and fate with the model BDE congeners. Concentrations of BDEs and PCBs were monitored in whole-fish tissues and liver tissues over the duration of the experiment. In addition, blood serum samples were taken and pooled among replicates to determine if any phenolic metabolites of BDE and PCBs were formed. Rapid assimilation of BDE 47 was observed relative to all other BDE and PCB congeners, whereas apparently no accumulation of BDE 99 occurred over the course of the experiment. Assimilation efficiencies for BDE 47 suggest that approximately 100% of the BDE 47 exposure was absorbed by carp tissues after 60 d. However, based on the time course of BDE 47 assimilation, it is improbable that all BDE 47 was assimilated; more likely, production of BDE 47 in carp tissues occurred as a result of debromination of higher-brominated compounds, possibly BDE 99. The net assimilation efficiencies of BDE 28 and BDE 153 were also apparently low (20 and 4%, respectively) relative to the three PCBs (40% assimilated) examined in this study. The low assimilation efficiency and high depuration rates for BDEs suggest a higher potential for biotransformation. While all three PCB compounds displayed very similar assimilation and depuration rates, three of the four BDE compounds displayed significantly different assimilation rates among BDE congeners and relative to the PCBs. This study suggests that BDEs have significantly different fate dynamics relative to PCBs in wild carp and likely other species of fish. PMID- 15352484 TI - Interactive effects of p,p'-dichlorodiphenyldichloroethylene and methoxychlor on hormone synthesis in largemouth bass ovarian cultures. AB - p,p'-Dichlorodiphenyldichloroethylene (DDE) and methoxychlor were tested alone and in combination to assess the similarity of their actions on hormone synthesis in gonadal tissue from largemouth bass (Micropterus salmoides floridanus), a species whose reproductive fitness has relevance to ecosystem health in Florida (USA). Gonads were harvested from adult female bass (age, two to three years) during the peak reproductive season (January-May), minced, and incubated in culture medium with or without test agents for 48 h. Duplicates of each treatment were performed in each of three experiments using tissue from a different female. Both 17beta-estradiol and testosterone were measured in aliquots of culture medium by validated radioimmunoassay procedures. Dose-response relationships of individual agents were characterized over a 6-log concentration range (1 X 10(-2) to 1 X 10(4) ppb). Both DDE and methoxychlor, tested individually, produced a dose-dependent decrease in testosterone levels. 17beta-Estradiol levels were unaffected. Mixtures of the agents were tested at all concentration combinations of 0.01, 1, 100, and 10,000 ppb in culture medium. Statistical tests indicated that of 16 dose combinations tested, 15 were antagonistic, and only 1 was additive based on the Loewe additivity model of no interaction. These results imply that methoxychlor and DDE inhibit testosterone production by different mechanisms in bass ovaries. PMID- 15352485 TI - Development of a whole-sediment toxicity test using a benthic marine microalga. AB - An acute whole-sediment toxicity test with a benthic marine microalga was developed and optimized using flow cytometry to distinguish algae (based on their chlorophyll a autofluorescence) from sediment particles. Of seven benthic marine algae screened, the diatom Entomoneis cf punctulata was most suitable because of its tolerance of a wide range of water and sediment physicochemical parameters, including salinity, pH, ammonia, and sulfide. A whole-sediment and water-only toxicity test based on inhibition of esterase activity in this species was developed. Enzyme activity rather than growth was used as the test endpoint, as nutrient release from sediments has previously been found to stimulate algal growth, potentially masking contaminant toxicity. The sensitivity of the bioassay to a range of metals (copper, zinc, cadmium, lead, arsenic, manganese) and phenol in water-only exposures was compared to the standard 72-h growth rate inhibition test. The esterase enzyme inhibition test was sensitive to copper, with a 3-h inhibitory concentration to cause a 50% (IC50) reduction in a fluorescein diacetate fluorescence value of 97 +/- 39 microg Cu/L. A concentration-dependent response was also observed in the presence of sediment particles (copper tailings), with and without dilution, using a control clean sediment. The primary route of exposure to copper was via pore water rather than by direct contact with tailings particles. This is the first whole-sediment bioassay developed with a benthic alga suitable for sediment quality assessment in marine/estuarine systems, and its advantages and limitations are discussed. PMID- 15352486 TI - Maternal transfer of selenium in Alligator mississippiensis nesting downstream from a coal-burning power plant. AB - Selenium (Se) is embryotoxic in many oviparous vertebrates, but little is known about maternal transfer of Se and its impact in reptiles. Over a four-year period, we collected three clutches of eggs of the American alligator (Alligator mississippiensis) from a single nest at a site contaminated with Se and compared egg and hatchling Se concentrations and clutch viability from this nest to nests downstream from the contaminated site (two clutches from two nests) and at a reference site (two clutches from two nests). Eggs and hatchlings from the nest at the Se-contaminated site and downstream nests had elevated Se concentrations (2.1-7.8 ppm) and lower viability (30-54%) compared to reference nests (1.4-2.3 ppm and 67-74% viability), but Se concentrations did not exceed reproductive toxicity thresholds established for other oviparous vertebrates. Selenium concentrations were higher in chorioallantoic membranes of eggs from Se contaminated sites, suggesting that this tissue may be useful as a nondestructive index of Se exposure for embryos of A. mississippiensis. Examination of these data suggests that further studies on uptake, accumulation, and reproductive success of crocodilian embryos exposed to excessive Se are warranted. PMID- 15352487 TI - Comparison of accumulation of micropollutants between igenous and transplanted zebra mussels (Dreissena polymorpha). AB - Zebra mussels (Dreissena polymorpha) were exposed at 12 canals and lakes situated in Flanders (Belgium), in cages for six weeks during the summer of 2002. Accumulation of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), hexachlorobenzene, and trace metals were measured in the transplanted mussels and levels compared to levels in indigenous mussels. Additionally, zebra mussels were exposed at a small lake in the vicinity of Antwerp (Belgium), and accumulation of contaminants was followed for an extended period from December 2001 to April 2002. Analysis of the pollutants in the indigenous mussels showed that the selected sites displayed a wide range of pollution from near to background to very high levels of metals and/or organic contaminants when compared to the literature. For organic contaminants and for most metals, comparison of levels between caged and resident mussels revealed no significant differences. Only for cadmium and nickel, significant differences were observed, with levels being either higher (cadmium) or lower (nickel) in caged mussels. For organic contaminants, significant correlations between levels in caged and resident mussels were found with r2 values up to 0.98. For some metals, no or poor correlations were found. At most sites, concentrations of those metals were of the same order of magnitude and comparable to levels in mussels from unpolluted sites. This might explain the absence of significant correlations. When mussels were exposed for an extended period, the concentration of some pollutants increased, whereas others decreased with time. Only in the case of certain metals did levels differ significantly because of the slow depuration of metals already present in the transplanted mussels. This is an additional indication that measured concentrations in transplanted mussels indeed reflected the local situation. With this study, we were able to prove the applicability of transplanted mussels as a biomonitoring tool for the accumulation of pollutants over a wide range of pollution. PMID- 15352488 TI - Runoff simulation with particle-associated azinphosmethyl in multispecies stream microcosms: implications for the field. AB - We investigated the acute (5 d) effects of particle-associated azinphosmethyl (AZP) in multispecies microcosms and assessed the results in the context to data obtained from a parallel field study undertaken in the Lourens River, South Africa. A runoff simulation was carried out in stream microcosms containing the macroinvertebrate fauna of an uncontaminated Lourens River site exposed to particle-associated AZP (control and 200, 1000, 5000, 20,000 microg/kg; three replicates each) for 1 h. Measured AZP concentrations in filtered microcosm water resulted in the following values: Not detectable (control) and 0.03, 0.2, 1.1, and 6.9 microg/L, respectively. The two highest treatments resulted in significantly (analysis of variance [ANOVA]) reduced total numbers of individuals, while the number of taxa was affected in the 20,000 microg/kg treatment only. A comparison with previous data suggests that observed effects partly resulted from particle-associated AZP. Particularly affected were six out of 14 macroinvertebrate taxa such as mayfly and stonefly taxa. In parallel, the distribution of macroinvertebrates at a pesticide-free and a contaminated stretch of the Lourens River was monitored five times during the spraying season in 2001 and 2002. Out of the 14 core taxa found in the microcosm study as well as in the field approach, 10 showed comparable reactions in the microcosm experiment and in their field distribution; they were either classified as affected or unaffected in both studies. Thus, we conclude that particle-associated AZP has the potential to affect the invertebrate community structure of the Lourens River and that microcosm studies employing field-relevant exposure scenarios may be valuable for a local risk assessment of pesticide-related community disruptions in the Lourens River. PMID- 15352489 TI - Destabilized lysosomes and elimination of polycyclic aromatic hydrocarbons and polychlorinated biphenyls in eastern oysters (Crassostrea virginica). AB - Contaminated oysters from near the Galveston Bay Ship Channel ([GBSC], TX, USA) were transplanted into clean aquariums in order to investigate the recovery of their lysosomal health. Destabilized lysosomes in the oyster hemocytes recovered from 81% to 34% after 23 d of elimination. Chemical analyses showed that transplanted oysters eliminated organic chemicals they had accumulated in their tissue. Enhanced lysosomal health (i.e., decreased lysosomal destabilization) was correlated positively with lower tissue concentrations of chemicals. This study indicates that lysosomal destabilization is a transient biomarker, which indicates only the current status (e.g., within one month) of chemical exposure and potential adverse biological effects. Analytical measurements revealed that polycyclic aromatic hydrocarbons (PAHs) generally were eliminated faster than polychlorinated biphenyls (PCBs) by a factor of 5 to 7. Body burdens of PCBs decreased exponentially, while the elimination pattern of PAHs were biphasic, representing a faster elimination during the early phase of elimination and, subsequently, a much slower elimination. This indicates that elimination of PAHs in the oysters is more complex and may include additional mechanisms compared to the uptake and elimination of PCBs. PMID- 15352490 TI - Plasma sex steroid concentrations and gonadal aromatase activities in African clawed frogs (Xenopus laevis) from South Africa. AB - Adult African clawed frogs (Xenopus laevis) were collected from a corn-growing region (CGR) and a non-corn-growing region (NCGR) with different exposure profiles for atrazine and related triazines. Physical, chemical, and biological parameters from the catchment areas were also measured. Frogs were surveyed for possible effects of exposure to triazine herbicides on plasma testosterone (T) and estradiol (E2) titers, gonadal aromatase activity, and gonad growth (GSI). Concentrations of both T and E2 varied among locations and were correlated to some accessory factors, such as pH, several ions, and metals. Greatest median plasma T concentrations (males: 19 ng/ml; females: 16 ng/ml) occurred in frogs inhabiting NCGR as compared to those from the CGR (males: 4 ng/ml; females: 1 ng/ml). Median E2 concentrations were also greater in frogs collected from the NCGR (males: 3 ng/ml; females: 28 ng/ml) than those in frogs from the CGR (males: 2 ng/ml; females: 5 ng/ml). Because some exposure to agricultural chemicals at both regions occurred, as did simultaneous exposures to multiple chemicals, a regression analysis was employed. Negative correlations were observed between plasma T concentrations and concentrations of atrazine, deisopropylatrazine, deethylatrazine, and tertbuthylazine in females and between T and diaminochlorotriazine in males. Estradiol in females exhibited a significant negative correlation with atrazine and deethylatrazine. No correlations were observed between gonadal aromatase activity or GSI and any of the agricultural chemicals measured. Median aromatase activities in ovaries varied among sampling sites ranging from 7 to >3000 times greater than those in males when measurable. Testicular aromatase activity was below the detection limit of the assay in male frogs at most of the sites. Although exposure to agricultural inputs did not affect aromatase activities, effects of atrazine or coapplied pesticides on sex steroid homeostasis cannot be excluded at this point. PMID- 15352491 TI - Association of heavy metals with metallothionein and other proteins in hepatic cytosol of marine mammals and seabirds. AB - Distribution of Cu, Zn, Cd, Ag, Hg, and Se were determined in hepatocytosol of northern fur seals (Callorhinus ursinus), black-footed albatrosses (Diomedea nigripes), and Dall's porpoises (Phocoenoides dalli). Copper, Zn, and Cd were accumulated preferentially in metallothionein (MT) fraction and their contents in MT fraction increased with the amounts in the hepatocytosol. Silver was bound to both high-molecular-weight substances (HMWS) and MT in the hepatocytosol for all three species, whereas the distribution of Ag in the cytosol was different among the three species. In northern fur seals, Ag mainly was bound to MT, whereas it mainly was associated with HMWS in Dall's porpoises. In contrast, Ag was distributed almost equally in both HMWS and MT for black-footed albatrosses. Mercury content in HMWS and Se content in HMWS and low-molecular-weight substances (LMWS) increased with their contents in hepatocytosol for all the three species. A significant positive correlation was found between Se and Hg contents in high-molecular weight (HMW) fraction in cytosol. The molar ratio of Hg and Se was close to unity in HMW fraction of the specimens with high Hg concentration in cytosol, implying that the Hg-Se complex was bound to the HMWS. Analysis of metals in the hepatocytosol by high-performance liquid chromatography/inductively coupled plasma-mass spectrometry (HPLC/ICP-MS) suggests that multiple isoforms of MT are present in hepatocytosol of the three species and that the metal profiles in hepatocytosols are different among the species. To our knowledge, this is the first report on the association of Ag with HMWS and MT in hepatocytosol of marine mammals and seabirds. Also, distribution and interaction of Hg and Se were investigated for the first time in hepatocytosol of the higher trophic marine animals. PMID- 15352492 TI - Baseline toxicity of a chlorobenzene mixture and total body residues measured and estimated with solid-phase microextraction. AB - Body residues of compounds with a narcotic mode of action that exceed critical levels result in baseline toxicity in organisms. Previous studies have shown that internal concentrations in organisms also can be estimated by way of passive sampling. In this experiment, solid-phase microextraction (SPME) fibers were used as a tool to estimate the body residues, which were then compared to measured levels. Past application of SPME fibers in the assessment of toxicity risk of samples has focused on separate exposure of fibers and organisms, often necessitated by the amount of agitation needed in order to achieve steady state in the fibers within a convenient time period. Uptake kinetic studies have shown that in SPME fibers with thin coatings, equilibrium concentrations can be reached without agitation within the time frame of a toxicity test. In contrast to toxicity experiments to date, the SPME fibers in the current study were exposed concomitantly to the test water with the organisms, ensuring an exposure under the exact same conditions. Fibers and two aquatic invertebrate species were exposed to a mixture of four chlorobenzenes with a narcotic mode of action. The total body residue of these compounds in the organisms was determined, as was the acute toxicity resulting from the accumulation. The total body residues of both species were correlated to the total concentrations in SPME fibers. It was concluded that toxicity could be predicted based on total body residue (TBR) estimates from fiber concentrations. PMID- 15352493 TI - Chronic toxicity and body residues of the nonpolar narcotic 1,2,3,4 tetrachlorobenzene in Chironomus riparius. AB - The use of internal concentrations as a dose parameter for baseline toxicity requires an understanding of the relationship between accumulation level and toxic effects, not only for acute but also for chronic exposure. In this study of chronic toxicity of the nonpolar narcotic 1,2,3,4-tetrachlorobenzene (TeCB) to Chironomus riparius, the chronic median lethal concentration (LC50) was determined to be 0.99 (0.54-1.82) microM, the median sublethal effect concentration (EC50) for growth was 0.76 (0.73-0.97) microM, and the chronic (sublethal) no-observed-effect concentration (NOEC) was 0.24 +/- 0.01 microM. An acute-to-chronic ratio of 9.8 was calculated from a previously determined acute LC50 value and this NOEC. The chronic critical body residue (CBR), 136 mmol/kg lipid, was the same as the acute CBR, previously determined. The similarity of the chronic and acute CBRs lends support to the exposure time independent aspect of baseline toxicity theory. An implication of this is that internal concentrations estimated by biomimetic sampling devices may be compared to acute CBR data to determine chronic baseline toxicity risk. Such sampling devices, solid-phase microextraction (SPME) fibers, were simultaneously exposed during the toxicity test. The results of this study suggest that body residues estimated with SPME may be used to predict baseline toxicity for various exposure durations. PMID- 15352494 TI - Influence of multigeneration acclimation to copper on tolerance, energy reserves, and homeostasis of Daphnia magna straus. AB - A multigeneration acclimation experiment was performed with Daphnia magna exposed to copper to assess possible changes in tolerance and to establish the optimal concentration range (OCEE) of this species. The hypothesis was tested that as the bioavailable background concentration of an essential metal increases (within realistic limits), the natural tolerance (to the metal) of the acclimated/adapted organisms and communities will increase. During 18 months the daphnids were exposed to six different, environmentally relevant, copper background concentrations ranging between 0.5 and 100 microg Cu L(-1) (7 x 10(-15) and 3.7 x 10(-9) M Cu2+). An increase in acute (effect concentration resulting in 50% immobility: 48-h EC50) and chronic copper (effect concentration resulting in 50% or 10% reproduction reduction: 21-d EC50, 21-d EC10) tolerance was observed with increasing exposure concentration. The 48-h EC50 increased significantly from 204 +/- 24 microg Cu L(-1) to 320 +/- 43 microg Cu L(-1). A nonsignificant change from 48.0 (47.9-48.0) microg Cu L(-1) to 78.8 (66.3-93.6) microg Cu L(-1) was noted in the chronic toxicity assays. The optimal concentration range was assessed using different biological parameters (i.e., net reproduction [R0]), energy reserves (Ea), body length measurements, filtration rates, and body burdens. After three generations of acclimation the OCEE ranged between 1 and 35 microg Cu L(-1) (2 X 10(-14) to 80 x 10(-12) M Cu2+). Body burden measurements revealed an active copper regulation up to 35 microg Cu L(-1) (80 pM Cu2+). It can be concluded that acclimation of D. magna to copper does occur in laboratory experiments, even at realistic copper background concentrations (10(-11) - 10(-9) M Cu2+). However, it is suggested that this phenomenon is of less importance in the context of regulatory risk assessments. An optimal copper concentration range for D. magna was observed between 1 and 35 microg Cu L(-1) (10(-14) - 10(-11) M Cu2+), indicating that copper deficiency can occur in routine laboratory cultures. PMID- 15352495 TI - Effects of chronic dietary copper exposure on growth and reproduction of Daphnia magna. AB - A matter of current, intense debate with regard to the effects of metals on biological systems is the potential toxicity of metals associated with food particles. Recently developed biotic ligand models (BLM), which predict the toxicity of waterborne metals, may not be valid if the dietary exposure route contributes to metal toxicity. The present study is, to our knowledge, the first that investigates the potential toxicity of dietary copper to a freshwater invertebrate (i.e., Daphnia magna) feeding on a live diet (i.e., the green alga Pseudokircheneriella subcapitata). Algae were exposed for 3 d to different copper concentrations, resulting in algal copper burdens between approximately 6.2 X 10( 16) and 250 x 10(-16) g cell(-1). These algae were then used as food in chronic, 21-d D. magna toxicity tests in which growth, reproduction, and copper accumulation were assessed. Three exposure scenarios were tested: A waterborne exposure, a dietary exposure, and a combined waterborne and dietary exposure. Although exposure to dietary copper resulted in an increased copper body burden of the adult daphnids, it did not contribute to toxicity and did not affect the 21-d effect concentrations expressed as waterborne copper, indicating that the previously established good predictive capacity of the chronic D. magna BLM is not affected. On the contrary, exposure to the highest dietary copper levels resulted in an increase of as much as 75% in growth and reproduction. To our knowledge, this is the first evidence that dietary copper exposure of a freshwater invertebrate feeding on a live diet resulted in a beneficial effect. PMID- 15352496 TI - Non-destructive rapid analysis of brominated flame retardants in electrical and electronic equipment using Raman spectroscopy. AB - Using Raman spectroscopy without any preparation, we analyzed the brominated flame retardant, polybromodiphenyl ethers (PBDEs), which has been prohibited by the European Union. The typical Raman bands of PBDEs, which are clearly different from those of other brominated flame retardants, are from 100 to 300 cm(-1). In our analysis, the detection limit was about 100 ppm, and the analysis took about 1 min, whereas the usual method of solvent extraction using gas chromatography/mass spectroscopy takes 50 h. PMID- 15352497 TI - Recent developments, characteristics, and potential applications of electrochemical biosensors. AB - The objective of this study is to analyze the technical importance, performance, techniques, advantages, and disadvantages of the biosensors in general and of the electrochemical biosensors in particular. A product of reaction diffuses to the transducer in the first generation biosensors (based on Clark biosensors). The mediated biosensors or second generation biosensors use specific mediators between the reaction and the transducer to improve sensitivity. The second generation biosensors involve two steps: first, there is a redox reaction between enzyme and substrate that is reoxidized by the mediator, and eventually the mediator is oxidized by the electrode. No normal product or mediator diffusion is directly involved in the third generation biosensors, direct biosensors. Based on the type of transducer, current biosensors are divided into optical, mass, thermal, and electrochemical sensors. They are used in medical diagnostics, food quality controls, environmental monitoring, and other applications. These biosensors are also grouped under two broad categories of sensors: direct and indirect detection systems. Moreover, these systems could be further grouped into continuous or batch operation. Therefore, amperometric biosensors and their current applications are focused on more in detail since they are the most commonly used biosensors in monitoring and diagnosing tests in clinical analysis. Problems related to the commercialization of medical, environmental, and industrial biosensors as well as their performance characteristics, their competitiveness in comparison to the conventional analytical tools, and their costs determine the future development of these biosensors. PMID- 15352498 TI - The interaction of copper-bipyridyl complex with DNA and cleavage to DNA. AB - The interaction of a copper-bipyridyl (bpy) complex with CT-DNA was investigated by voltammetry, absorption and fluorescence spectrophotometry. The binding constant of the Cu(bpy)2(2+) complex interacting with DNA was 3.24 x 10(4) L/mol and the ability binding of Cu(bpy)2(2+) to DNA was 1.3-times as large as that of Cu(bpy)2+ to DNA. DNA could be efficiently cleaved by a potential-modulated method in the presence of the Cu(bpy)2(2+) complex. The fragments of the cleaved DNA were separated by high-performance liquid chromatography (HPLC). The experimental results revealed that the proposed method for DNA cleavage is highly efficient. PMID- 15352499 TI - Attachment of DNA to the carbon fiber microelectrode via gold nanoparticles for simultaneous determination of dopamine and serotonin. AB - A novel biochemical sensor was fabricated on a carbon fiber microelectrode, which consisted of an inner layer of electrodeposited gold nanoparticles, as a nano array electrode, and an outer layer of electrodeposited calf thymus ds-DNA at +1.5 V vs. SCE. This modified electrode was characterized by X-ray photoelectron spectroscopy, scanning electron microscope, atomic force microscopy, cyclic voltammetry and differential pulse voltammetry (DPV). It was found that this electrochemical sensor exhibits a strong catalytic activity toward the oxidation of dopamine (DA), serotonin (5-HT) and ascorbic acid (AA), as a result of resolving the anodic voltammetric peaks of DA, 5-HT and AA into three well defined peaks. Simutaneous DPV determination of DA and 5-HT can be achieved in the presence of 2000-fold AA. The modified electrode shows good sensitivity, selectivity and stability. PMID- 15352500 TI - A novel membrane sensor for histamine H1-receptor antagonist "fexofenadine". AB - The construction and general performance of thirteen new polymeric membrane sensors for the determination of fexofenadine hydrochloride based on its ion exchange with reineckate, tetraphenylborate and tetraiodomercurate have been studied. The effects of membrane composition, type of plasticizer, pH value of sample solution and concentration of the analyte in the sensor internal solution have been thoroughly investigated. The novel sensor based on reineckate exchanger shows a stable, potentiometric response for fexofenadine in the concentration range of 1 x 10(-2) - 2.5 x 10(-6) M at 25 degrees C that is independent of pH in the range of 2.0 - 4.5. The sensor possesses a Nernstian cationic slope of 62.3+/ 0.7 mV/concentration decade and a lower detection limit of 1.3 x 10(-6) M with a fast response time of 20 - 40 s. Selectivity coefficients for a number of interfering ions and excipients relative to fexofenadine were investigated. There is negligible interference from almost all studied cations, anions, and pharmaceutical excipients, however, citrizine that has a structure homologous to that of fexofenadine was found to interfere. The determination of fexofenadine in aqueous solution shows an average recovery of 99.83% with a mean relative standard deviation (RSD) of 0.5%. Direct potentiometric determination of fexofenadine in tablets gave results that compare favorably with those obtained by standard spectrophotometric methods. Potentiometric titration of fexofenadine with phosphomolybdic acid as a titrant has been monitored with the proposed sensor as an end point indicator electrode. PMID- 15352501 TI - Amperometric glucose biosensor based on mediated electron transfer between immobilized glucose oxidase and plasma-polymerized thin film of dimethylaminomethylferrocene on sputtered gold electrode. AB - We propose an electron transfer-mediated amperometric enzyme biosensor based on plasma-polymerized thin film of dimethylaminomethylferrocene (DMAMF) on a sputtered gold electrode. The DMAMF plasma-polymerized film is deposited directly onto the surface of the electrode under dry conditions. The resulting thin film not only has redox sites but also is extremely thin (approximately 20 nm), adheres well onto the substrate (electrode), has a flat surface and a highly crosslinked network structure, and is hydrophilic in nature. Glucose oxidase is densely immobilized onto the surface of DMAMF plasma-polymerized film on the gold electrode. From the electrochemical measurement, the biosensor can cover the wide range of glucose concentration (1.3 - 81 mM) at +350 mV of applied potential. The current response of the glucose biosensor was decreased by less than 5% in an aerobic solution as compared to that in an anaerobic solution. These show that the DMAMF plasma-polymerized films play a role as the electron transfer mediators between the reaction center of enzyme and the electrode. PMID- 15352502 TI - Characterization of the rutin-metal complex by electrospray ionization tandem mass spectrometry. AB - According to the strong application background of bioflavonoid and metal flavonoid complexes, novel electrospray ionization tandem mass spectrometry (ESI MSn) was applied to investigate the structure and fragmentation mechanism of transition metal-rutin complexes. In the full-scan mass spectra, different stoichiometric ratios of rutin-metal complexes were found. In the reaction between rutin and Cu, four kinds of complexes with four different stoichiometric ratios were produced. In the reaction between rutin and Zn, Mn(II), and Fe(II), only two kind of complexes with stoichiometric ratios of 1:1 and 1:2 occured. In further tandem mass spectrometric experiments of different rutin-metal complexes, product fragments came from the neutral loss of the external rhamnose and the internal glucose unit, oligosaccharide chain, aglycone, and small organic molecules. According to the MSn data, we proposed a mechanism for all fragments of the rutin-Cu complex A and the structure of two rutin-Cu complexes, C and D. PMID- 15352503 TI - Oxygen indicator composed of an organic/inorganic hybrid compound of methylene blue, reductant, surfactant and saponite. AB - An organic/inorganic hybrid compound consisting of methylene blue, a cationic surfactant and a reductant intercalated into saponite was found to serve as an oxygen indicator that changes color in the presence of oxygen. A mixture of a blue colored dye, methylene blue, a reductant in the form of ascorbic acid or reducing sugar, and cetyltrimethylammonium ion intercalated into synthetic saponite became colorless in an atmosphere having an oxygen concentration of less than 0.1 vol%, and then returned to its blue color as a result of subsequent exposure to air. An oxygen indicator, in the form of a thin film coated on paper prepared by adding a pigment, phloxine B, to the above organic/inorganic hybrid compound, exhibited a pink color at oxygen concentrations of less than 0.1 vol%, and a blue color at oxygen concentrations of higher than 0.5 vol%. In addition, this oxygen indicator exhibited superior photo-fading resistance and storage stability compared with indicators using only methylene blue as the functional dye. PMID- 15352504 TI - Field determination of trace iron in fresh water samples by visual and spectrophotometric methods. AB - Sensitive visual and micro spectrophotometric methods have been developed for field determination of trace iron in fresh water samples. For the visual method, a water sample (0.45-microm filtrate acidified to 0.1 M HCl) was placed in a glass vial and mixed with a reagent solution containing 1,10-phenanthroline, sodium thiocyanate and 0.1 M HCl. Iron was extracted as pink ferroin thiocyanate with 1 ml of 4-methy-2-pentanone. The sample up to 20 ml was added step-by-step, until the color of the extract was detected visually. Without any special instrument or color standard, iron down to 0.001 mg 1(-1) (0.025 microg) in a sample can be determined with an error of 20% in the field. For the micro spectrophotometric method, the extract for 20 ml of sample was separated by capillary suction in a column (micro pipette chip) with acrylic fibers. A part of the extract was pushed out into a micro cell for the absorbance measurement at 525 nm. The column was re-usable after washing with ethanol. This method had a detection limit of 0.001 mg 1(-1) and allowed determinations within an error of 5%. The proposed methods were applied to deionized-, tap-, river-, lake- and reservoir-water samples. PMID- 15352505 TI - Neutral-carrier-type potassium ion-selective electrodes based on polymer supported liquid-crystal membranes for practical use. AB - Polymer-supported liquid-crystal membranes have been designed for neutral-carrier type potassium ion-selective electrodes, aiming for practical applications of high-performance liquid-crystalline membrane ion sensors. Two types of polymer supported liquid-crystal membranes were tested for their usefulness; one is microporous poly(tetra fluoroethylene) (PTFE) membranes impregnated by thermotropic liquid-crystalline compounds, and another is poly(methyl methacrylate) (PMMA) membrane dispersing the same liquid-crystalline compounds. Both of the polymer-supported liquid-crystal membranes containing a liquid crystalline benzo-15-crown-5 neutral carrier as well as a lipophilic anion excluder work well as ion-sensing membranes for potassium ion-selective electrodes, the ion selectivities of which can be switched by the measurement temperatures. Specifically, PTFE-impregnated liquid-crystal membranes are better than the PMMA-dispersed ones in the sensitivity and selectivity of the resulting ion electrodes. A potassium ion assay in blood sera has proved that neutral carrier-type ion-selective electrodes based on the polymer-supported liquid crystal membranes are reliable for practical uses. PMID- 15352506 TI - Activation effects of a platinum electrode by laser pulse irradiation on the electro-oxidation of glucose in alkaline solution. AB - In order to demonstrate the activation effects of a Pt electrode by laser pulse irradiation, the electro-oxidation of glucose was tested at an activated Pt electrode by cyclic voltammetry. A fixed potential was applied to the electrode, and then the electrode was irradiated with laser pulses from a Nd:YAG laser at 20 Hz for 20 s. Activation by the laser pulse irradiation gave two remarkable effects on cyclic voltammograms from the electro-oxidation of glucose in a 0.1 mol dm(-3) NaOH solution, i.e., surface modulation and cleaning effects. Significant differences were found in the cyclic voltammograms at the activated and at the simply polished electrodes. Such differences in the oxidation waves are attributed to a crystallographic change of the electrode surface induced by a laser ablation, accompanied by laser pulse irradiation. Due to the cleaning effect, the activated Pt electrode gave a sharp oxidation wave at -0.3 V even in real samples containing various organic compounds that could foul the electrode, though the activated Pt electrode lacked selectivity to the electro-oxidation of glucose. PMID- 15352507 TI - Electrochemical reduction of 1-([(4-halophenyl)imino]methyl)-2-naphthols in aprotic media. AB - The electrochemical reduction of 1-([(4-halophenyl)imino]methyl)-2-naphthols on graphite electrodes was studied using cyclic voltammetry, chronoamperometry, constant-potential coulometry and preparative constant-potential electrolysis techniques. The data revealed that the reduction on graphite was irreversible and followed an EC mechanism. The diffusion coefficients and the number of electrons transferred were determined using the chronoamperometric Cottrell slope and the ultramicro disc Pt-electrode steady-state current. The number of electrons was also determined by bulk electrolysis. The compounds were subjected to constant potential preparative electrolysis and the electrolysis products were purified and identified by spectroscopic methods. Based on these findings, a mechanism for the electro-reduction process is proposed. PMID- 15352508 TI - Highly selective thiocyanate electrode based on bis-[N-(2 hydroxyethyl)salicylaldimino]copper(II) complex as a neutral carrier. AB - A novel selective thiocyanate electrode of a PVC membrane based on bis-[N-(2 hydroxyethyl)salicylaldimino]copper(II) [Cu(II)-BNSDM] as a neutral carrier is reported. The selectivity sequence of this electrode is as follows: SCN- > ClO4- > I- > Sal- > Br- > NO2- > Cl- > NO3- > H2PO4- > SO4(2-), which is an anti Hofmeister selectivity sequence. The electrode exhibits a Nernstian potential linear range to thiocyanate from 1.0 x 10(-1) to 6.0 x 10(-6) mol/L with a detection limit of 2.0 x 10(-6) mol/L and a slope of (-59.0+/-0.2)mV/decade in pH 5.0 of a phosphate buffer solution at 25 degrees C. Electrochemical impedance spectroscopy (EIS) and UV spectroscopy techniques were performed to understand the response mechanism of the electrode. The electrode had the advantage of simplicity, high selectivity, fast response and low detection limit compared with the other electrode, which had been reported before. The electrode had been applied to wastewater analysis, and the obtained results were satisfactory. PMID- 15352509 TI - Low-capacity cation-exchange chromatography of amino acids using a novel sulfoacylated macroreticular polystyrene-divinylbenzene column with binary gradient elution. AB - This paper describes a versatile technique for amino-acid separation using a novel low-capacity sulfoacylated macroreticular polystyrene-divinylbenzene cation exchange column with a simple binary high-pressure pH gradient elution. Proteinic 16 amino acids were well separated within 50 min using a H3PO4/Na2HPO4-CH3CN eluent system, and the cycle time was about 70 min. The chromatography with postcolumn OPA fluorescent detection was reproducible with RSDs less than 1% for retention times, and was quantitative with RSDs less than 5% for area responses. A linear regression line with an r2 value above 0.9990 was obtained for each analyte in concentration from 0.1 to 10 microM by 20 microL injection. The method was applicable to the separation and detection of urinary diagnostic amino acid due to inborn errors of metabolism, such as phenylketonuria. The analytical costs would be decreased by using the proposed method. PMID- 15352510 TI - Liquid chromatography of uranium complexes of tetradentate Schiff bases. AB - Dioxouranium together with copper(II), nickel(II) and iron(II) were extracted in chloroform as complexes of bis(salicylaldehyde)-dl-stilbenediimine (dl-H2SA2S) or bis(salicylaldehyde)-meso-stilbenediimine (meso-H2SA2S), and separated by liquid chromatography with UV detection. The linear calibration range and detection limits were 40 - 200 ng and 10 ng/injection for each metal ion. The method was applied to the determination of uranium from mineral ore samples at concentrations of 30 - 700 microg/g with coefficients of variation from 3.6 to 5.5%. The relative elution of dioxouranium complexes of different Schiff bases was examined from reversed-phase HPLC; the substitution of methyl and phenyl groups at the bridge position enhanced the column retention of uranyl complexes. PMID- 15352511 TI - Simultaneous kinetic-spectrophotometric determination of hydrazine and acetylhydrazine in micellar media using the H-point standard addition method. AB - The H-point standard addition method (HPSAM), based on a spectrophotometric measurement for the simultaneous determination of hydrazine and acetylhydrazine, is described. This method is based on the difference between the rates of their reactions with N,N-dimethylaminobenzaldehyde (DAB) in the presence of sodium dodecyl sulfate (SDS) in acidic media. The results showed that hydrazine and acetylhydrazine could be determined simultaneously in the range of 0.020 - 0.70 and 0.20 - 5.0 mg L(-1), respectively. Under the working conditions, the proposed method was successfully applied to the simultaneous determination of hydrazine and acetylhydrazine in several synthetic mixtures and plasma and water samples. PMID- 15352512 TI - A sequential injection system for the spectrophotometric determination of calcium, magnesium and alkalinity in water samples. AB - A sequential injection methodology for the spectrophotometric determination of calcium, magnesium and alkalinity in water samples is proposed. A single manifold is used for the determination of the three analytes, and the same protocol sequence allows the sequential determination of calcium and magnesium (the sum corresponds to the water hardness). The determination of both metals is based on their reaction with cresolphtalein complexone; mutual interference is minimized by using 8-hydroxyquinoline for the determination of calcium and ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) for the determination of magnesium. Alkalinity determination is based on a reaction with acetic acid, and corresponding color change of Bromcresol Green. Working ranges of 0.5 - 5 mg dm(-3) for Ca, 0.5 - 10 mg dm(-3) for Mg, and 10 - 100 mg HCO3- dm( 3), for alkalinity have been achieved. The results for water samples were comparable to those of the reference methods and to a certified reference water sample. RSDs lower than 5% were obtained, a low reagent consumption and a reduced volume of effluent have been accomplished. The determination rate for calcium and magnesium is 80 h(-1), corresponding to 40 h(-1) per element, while 65 determinations of alkalinity per hour could be carried out. PMID- 15352513 TI - Identification of glass and ceramics by X-ray fluorescence analysis with a pyroelectric X-ray generator. AB - Applications of X-ray fluorescence (XRF) analysis with a pyroelectric X-ray generator are presented. Glass and ceramics were analyzed with this novel X-ray generator to examine its capability for analyzing nonmetallic inorganic material. Although the power of X-ray output was a few orders of magnitude lower than conventional X-ray tubes, many elements such as Si, K, Ca, Ti, Cr, Fe, Zn, Sr, Ba, and Pb were detected in glass and ceramic samples. Light elements such as Na, Mg, and Al were not detected because of the low fluorescence yield and the absorption of fluorescent X-rays in air. The elements detected or the relative peak intensities of the elements were different from each other, and the samples analyzed were identified by the XRF spectrum, notwithstanding the low power of the X-ray output. This novel device showed the sufficient capability for preliminary screening before strict identification of analytes. The pyroelectric X-ray generator can also be used to analyze large samples that cannot be put into ordinary XRF spectrometers because the device has no dimensional limitation of analytes. PMID- 15352514 TI - Single-channel recordings of gramicidin at agarose-supported bilayer lipid membranes formed by the tip-dip and painting methods. AB - Agarose-supported BLMs were prepared by the tip-dip and painting methods, and single-channel recordings of gramicidin were examined for the development of an ion-channel sensor. The supported BLMs formed by the tip-dip method had an electric resistance of >1.0 x 10(11) omega and a longer lifetime as compared with unsupported ones, which enabled single-channel recordings of gramicidin. The supported BLMs formed by the painting method also enabled single-channel recordings, but the lifetime was shorter than that of unsupported planar BLMs formed by the monolayer folding method. PMID- 15352515 TI - Analysis of self-assembled monolayers by thermal desorption mass spectrometry: neighborhood interaction and hydrogen/deuterium exchange. AB - Two ideas for characterizing the local structure in mixed self-assembled monolayers (SAMs) were introduced. First, to use thermal desorption mass spectrometry (TD-MS) to probe nearest neighborhoods in the mixed monolayers. Second, to use hydrogen/deuterium exchange as a probe of the accessibility of acidic protons to exchange. Neighborhood interactions provided an opportunity to observe microscopic phase changes at the SAMs surface as a function of the OH terminal ratio. H/D exchange between surficial OH-terminal SAMs and D2O was successfully observed. H/D exchange was of importance in characterizing the surface properties and molecular basis interactions for the design of bio interface structures using self-assembled monolayers. PMID- 15352516 TI - Spectrophotometric and AAS determination of trace amounts of cobalt after preconcentration by using alpha-benzilmonoxime-microcrystalline naphthalene. AB - The formed cobalt-a-benzilmonoxime complex was adsorbed onto microcrystalline naphthalene. Then it was determined by zero and first derivative spectrophotometry and by atomic absorption spectrophotometry (AAS) after dissolving into chloroform and methylisobutylketone (MIBK), respectively. Under optimum conditions, cobalt in the range of 1.0 - 20.0, 0.4 - 30.0 and 2.5 - 50.0 microg could be determined by spectrophotometry, first derivative spectrophotometry and AAS method, respectively. By the method, a preconcentration factor equal to approximately 30 for cobalt was obtained. The effect of diverse ions on the determination of 5.0 microg cobalt was also studied. The method was successfully applied to some pharmaceuticals and synthetic alloy samples. PMID- 15352517 TI - Validated spectrophotometric methods for the determination of amiodarone hydrochloride in commercial dosage forms using p-chloranilic acid and 2,3 dichloro-5,6-dicyano-1,4-benzoquinone. AB - Two simple, sensitive and economical spectrophotometric methods have been developed for the determination of amiodarone hydrochloride in pure form and commercial dosage form. These methods (A and B) are based on the reaction of amiodarone base as n-electron donor with p-chloranilic acid and 2,3-dichloro-5,6 dicyano-1,4-benzoquinone (DDQ) as pi-acceptors to give highly colored complex species which absorb maximally at 535 and 570 nm, respectively. Beer's law is obeyed in the concentration ranges 10.0 - 360.0 and 2.0 - 65.0 microg ml(-1) for methods A and B, respectively. Application of the proposed methods to commercial pharmaceutical tablets are presented. PMID- 15352518 TI - Spectrofluorimetric determination of trace amounts of europium(III) ion with lutetium(III)-sparfloxacin-sodium dodecyl sulfate luminescence enhancement system. AB - A new luminescence-enhancement system based on complex formation between europium and sparfloxacin in the presence of lutetium in a sodium dodecyl sulfate solution has been discovered. By adding a suitable amount of Lu3+ to the Eu-sparfloxacin sodium dodecyl sulfate system, the luminescence can be enhanced by approximately 5-fold compared with that of the system without Lu3+. Under the optimum conditions, the luminescence intensity of the system is a linear function of the concentration of europium in the range of 1.0 x 10(-10) - 5.0 x 10(-7) mol L(-1). The detection limit of europium is 2.0 x 10(-13) mol L(-1) (S/N = 3). The system was used for the determination of trace amounts of europium in rare earth samples with satisfactory results. PMID- 15352519 TI - Identification of two novel interferon-stimulated genes from cultured CAB cells induced by UV-inactivated grass carp hemorrhage virus. AB - Interferon (IFN) exerts its antiviral effect by inducing the expression of a number of IFN-stimulated genes (ISGs) to establish a host antiviral state. Earlier studies identified some important fish IFN system genes from IFN-induced CAB cells (crucian carp Carassius auratus L. embryonic blastulae cells) after treatment with UV-inactivated GCHV (grass carp hemorrhage virus). Herein, the cloning of 2 novel IFN-stimulated genes, termed Gig1 and Gig2, is described for the same cell system. The complete cDNA sequences of Gig1 and Gig2 contain 1244 bp encoding for a 194-amino-acid protein and 693 bp for a 158-amino-acid protein, respectively. A search of public databases revealed that these are 2 novel IFN stimulated genes, since neither significant homologous genes nor conserved motifs were identified. Active GCHV, UV-inactivated GCHV and CAB IFN-containing supernatant (ICS) induced transcription of these genes and distinct kinetics were observed. An analysis of differences in expression between the 2 genes and the IFN signal factors CaSTAT1 and CaIRF7 indicated that GCHV infection activated different signal pathways for their up-regulation. Upon virus infection, the transcription of Gig1 but not of Gig2 is strongly suppressed by cycloheximide (CHX). In contrast, following treatment with CAB IFN-containing supernatant, CHX does not inhibit either gene transcription. The results suggest that GCHV infection can induce expression of both Gig1 and Gig2 via newly synthesized CAB IFN, most probably through the JAK-STAT signal pathway, and can also directly activate Gig2 transcription without ongoing protein synthesis. PMID- 15352520 TI - Protection of atlantic salmon Salmo salar against infectious pancreatic necrosis after DNA vaccination. AB - Although vaccines against infectious pancreatic necrosis (IPN) based on inactivated virus or recombinant structural viral proteins are commercially available, the protection is not complete and the disease is still a problem for the Atlantic salmon Salmo salar farming industry. In the present study, 5 different plasmids that expressed whole or parts of the large open reading frames (ORF) of Segment A of the IPN virus (IPNV) were constructed. The plasmids were shown to express proteins in cell cultures and in zebrafish Danio rerio in vivo. The specificities of the expressed proteins were confirmed by staining with IPNV specific monoclonal antibodies (MAb) The plasmids were then used alone or in different combinations to vaccinate groups of Atlantic salmon, which subsequently were challenged in an experimental assay for IPN. A high level of protection was induced only by the plasmid combination that contained a plasmid expressing all the large ORF polyprotein. PMID- 15352521 TI - Vibrio parahaemolyticus and V. harveyi cause detachment of the epithelium from the midgut trunk of the penaeid shrimp Sicyonia ingentis. AB - Shrimp Sicyonia ingentis were either injected with Vibrio parahaemolyticus (10(4) CFU) or V. harveyi (10(6) CFU) or immersed in ASW containing either species at 10(5) CFU ml(-1). These densities were shown in preliminary experiments to kill approximately half the population by 7 d. On Day 7, surviving shrimp were classified as either diseased or apparently healthy, and their midgut trunks (MGT) were examined by light and electron microscopy. All shrimp immersed in ASW containing either species of Vibrio showed detachment of the epithelium in the MGT. In shrimp injected with either species of Vibrio, epithelial detachment was common in diseased shrimp but not in apparently healthy animals. Experiments with live shrimp were supported by in vitro experiments where MGTs were removed, tied off at both ends, and injected with either pathogenic bacteria (V. parahaemolyticus or V. harveyi), non-pathogenic bacteria (Bacillus subtilis or Escherichia coli), or ASW. After 2 h incubations in ASW at 15 degrees C, the MGTs were processed and examined. The epithelium consistently detached from isolated MGTs injected with either species of Vibrio, but not from MGTs injected with non pathogenic bacteria or ASW. Because the MGT epithelium secretes the peritrophic membrane, loss of the epithelium eliminates 2 layers that may restrict penetration of ingested pathogens into the shrimp body and may disrupt the osmoregulatory function of the MGT. A second finding was that fixed, large granule hemocytes associated with the basal lamina degranulated in the presence of the 2 species of Vibrio, but not with the non-pathogenic bacteria or ASW. These blood cells may help fight specific bacteria penetrating the MGT. PMID- 15352522 TI - Impact of PCB on resistance to Flavobacterium psychrophilum after experimental infection of rainbow trout Oncorhynchus mykiss eggs by nanoinjection. AB - The effects of sublethal exposure of a commercial blend of polychlorinated biphenyls (PCB), i.e. Clophen A50, on disease resistance to the aetiological agent of rainbow trout fry syndrome, Flavobacterium psychrophilum, were investigated. Newly fertilised rainbow trout Oncorhynchus mykiss eggs were nanoinjected with 2 doses of Clophen A50 (0.4 or 2 microg egg(-1)) and/or 100 colony forming units of F. psychrophilum. The mean cumulative mortality in control groups, and groups exposed to the lower dose of Clophen A50 (0.4 microg egg(-1)) was below 5.0%. The mean cumulative mortality in groups exposed to the higher dose of Clophen A50 (2.0 microg egg(-1)) was 5.8%, which was not significantly different from the control groups. In all groups infected with F. psychrophilum, with or without exposure to Clophen A50, significantly higher cumulative mortalities compared with control groups were recorded. No differences in mortality were recorded between groups exposed to bacteria alone or bacteria in combination with the higher dose of Clophen A50 (21.6 and 20.4%, respectively). Decreased disease resistance was recorded in groups exposed to F. psychrophilum and the lower dose of Clophen A50, with a mean cumulative mortality of 56.0%. These results could be due to non dose-dependent effects on the immune system, or toxic effects of PCB or their metabolites on the bacteria in groups exposed to the higher dose of Clophen A50. The present study indicates that maternal transfer of PCB might affect disease resistance to vertically transmitted F. psychrophilum. PMID- 15352523 TI - Some immune parameters in carp cyprinus Carpio susceptible and resistant to the haemoflagellate Trypanoplasma borreli. AB - The present study addresses aspects of the (specific) immune response of carp to the haemoflagellate Trypanoplasma borreli. Sera of resistant carp contained antibodies, which agglutinated the flagellates in vitro. When flagellates were incubated in fish sera from resistant carp, binding of antibodies to flagellates could be demonstrated by flow cytometry, and T. borreli were effectively killed. Heat-treatment of the sera prevented killing, indicating that complement activation is important for the control of a T. borreli infection. Sera of carp that were highly susceptible to infection with T. borreli contained no antibodies capable of binding to or killing the parasite. Furthermore, specific antibodies were not generated after experimental infection. This lack of antibody production in susceptible carp is not due to a general unresponsiveness of lymphoid cells, since peripheral blood leukocytes (PBL) from susceptible and resistant carp responded to mitogenic stimuli in vitro with lymphocyte proliferation in a similar manner. However, viable flagellates were significantly less able to stimulate proliferation of PBL from susceptible carp. In vitro-produced culture supernatants of freshly isolated PBL from both carp lines (but not those of head kidney cells) differentially modulated the mitogen-induced proliferation of PBL from susceptible and resistant carp. The supernatants enhanced the proliferation of leukocytes obtained from individuals from the same carp line, but suppressed the mitogen-induced proliferation of PBL from the other line tested. This indicates that lymphoid cells from susceptible and resistant carp differ in their spectrum of spontaneously produced immunomodulatory mediators. Whether this is decisive for a T. borreli-specific and successful immune response is discussed. PMID- 15352524 TI - Ultrastructure of Spironucleus salmonis n. comb. (formerly Octomitus salmonis sensu Moore 1922, Davis 1926, and Hexamita salmonis sensu Ferguson 1979), with a guide to spironucleus species. AB - Diplomonad flagellates can be associated with significant morbidity and mortality in fishes, particularly in farmed salmonids. Diagnosis using only light microscopy is limited, and species cannot be confirmed. We therefore undertook a comprehensive transmission electron microscopy study of 20 trophozoites from the intestine of farmed juvenile rainbow trout Oncorhynchus mykiss from Northern Ireland (reported as Hexamita salmonis by Ferguson in 1979). Re-assignment to the genus Spironucleus was determined based on the anteriorly tapering and intertwined elongate nuclei, anterior-medial kinetosomes, and the presence of a flagellar pocket. At the species level we observed a tri-radiate pattern of microtubules in the opening of the asymetrical striated lamina, comprising 3 to 5 microtubules following the edge of the flagellar pocket, 3 radiating away from the opening of the striated lamina, and 4 to 5 curving over the striated lamina. We observed electron-dense plaques adjacent to the kinetosomes, electron-dense bodies, numerous free ribosomes, aggregations of glycogen, bowl-shaped membranous structures, rough endoplasmic reticulum, and a novel distinctive pyriform sac of densely packed free ribosomes at the posterior of the cell (8-shaped in transverse section, and delineated by furrowed endoplasmic reticulum). We now propose to rename H. salmonis sensu Ferguson (1979) Spironucleus salmonis. Our review of the morphology of Octomitus salmonis illustrated by Moore (1922a,b) and Davis (1926), has shown that this organism also belongs to the genus Spironucleus. We synonymise H. salmonis sensu Ferguson (1979) with O. salmonis sensu Moore (1922) and Davis (1926), and rename them S. salmonis. An expanded diagnostic guide, including new cytoplasmic elements, is presented for the 4 species of Spironucleus from fishes (S. barkhanus, S. salmonis, S. torosa and S. vortens). We recommend that type descriptions of diplomonads be based on organisms taken directly from the host rather than from in vitro culture in order to reliably include the cytoplasmic organelles. PMID- 15352525 TI - 18S ribosomal DNA-based PCR identification of Neoparamoeba pemaquidensis, the agent of amoebic gill disease in sea-farmed salmonids. AB - Neoparamoeba pemaquidensis is a parasomal amoeboid protozoan identified as the agent of amoebic gill disease (AGD) in Atlantic salmon Salmo salar reared in sea pens in Tasmania, Australia, and coho salmon Oncorhynchus kisutch farmed on the west coast of the USA. Outbreaks of AGD caused by immunologically cross-reactive paramoebae have also been reported in sea-farmed salmonids in several other countries. Complete 18S rDNA sequences were determined for respective paramoebae isolated from infected gills of salmon from Tasmania and Ireland, and N. pemaquidensis isolates from the USA and UK, including representative free-living isolates. Alignments over 2110 bp revealed 98.1 to 99.0% sequence similarities among isolates, confirming that paramoebae implicated in AGD in geographically distant countries were homologous and belonged to the same species, N. pemaquidensis. The results supported previous findings that N. pemaquidensis exists as a widely distributed, amphizoic marine protozoan. Partial 18S rDNA sequences were obtained for the ultrastructurally similar species, N. aestuarina, and for the morphologically similar but non-parasomal amoeba Pseudoparamoeba pagei. N. aestuarina had 95.3 to 95.7% sequence similarities with N. pemaquidensis strains, which distinguished 2 closely related but separate species. Neoparamoeba spp. were not analogous to P. pagei or to other marine Gymnamoebia. We designed 4 oligonucleotide primers based on elucidated 18S rDNA sequences and applied them to single-step and nested 2-step PCR protocols developed to identify N. pemaquidensis to the exclusion of apparently closely related and non-related protistan taxa. Nested PCR was able to detect the AGD parasite from non-purified, culture-enriched net microfouling samples from Atlantic salmon sea-pens in Tasmania, and confirmed that N. pemaquidensis was also responsible for AGD in chinook salmon O. tshawytscha in New Zealand. Our sequence and PCR analyses have now shown that AGD affecting 3 different salmonid species farmed in 4 countries are associated with N. pemaquidensis. A species specific diagnostic PCR provides for the first time, a highly specific detection and identification assay for N. pemaquidensis that will facilitate future ecological and epidemiological studies of AGD. PMID- 15352526 TI - 'Cup cell disease' in the colonial tunicate Botryllus schlosseri. AB - A new progressive, fatal disease called 'cup cell disease' was characterized in ex situ cultures of Botryllus schlosseri, a colonial tunicate. The disease originated as a few dark spots growing within zooids. The infected colonies then started to deteriorate, morphologically diagnosed by ampullar retraction, lethargic blood circulation and by a swollen and soft tunic matrix. In later stages of the disease, developed buds were also affected. Many large black dots were scattered within the tunic matrix, and zooids were transformed to opaque, dilated, sac-like structures, signaling impending death. Colonies were infected periodically, even without direct tissue contact. The time course from first appearance to colony death ranged between 30 and 45 d. Histological studies, in vitro culturing of blood cells and blood smears revealed the existence of numerous cup-like cells (up to 4.8 microm diameter on average) with a yellowish cell wall and transparent cytoplasm that was not stained by various dyes (except azocarmine-G). Cells were refractive under bright-field illumination and revealed a flattened wall with flanges, characteristic of species of the phylum Haplosporidia. Cup cells aggregated in blood vessels and in internal parts of zooids and buds and were phagocytosed by blood cells. In a single case, plasmodia like structures were found only in the tunic matrix of an infected colony. This is the first record in botryllid ascidians of an infectious lethal disease associated with haplosporidian protists. PMID- 15352527 TI - Cloning and sequencing of envelope proteins (VP19, VP28) and nucleocapsid proteins (VP15, VP35) of a white spot syndrome virus isolate from Korean shrimp. AB - Since our first report in 1998, white spot syndrome virus (WSSV) has become wide spread on the southern and western coasts of Korea. Almost all shrimp in ponds die within 3 to 4 d after the first dead shrimp are observed with gross lesions ranging from abnormal red body discoloration to white spots in the cuticle. From one isolate, we cloned and sequenced WSSV genomic DNA coding for VP19 and VP28 envelope proteins and VP15 and VP35 nucleocapsid proteins. Putative protein sequences were submitted to GenBank and assigned accession numbers AY316119 (VP19), AY324881 (VP28), AY374120 (VP15) and AY325896 (VP35). At the nucleotide level, VP19, VP28 and VP15 sequences were, respectively, 99, 100 and 100% identical to those of China, Indonesia, Japan and the United States and the VP35 sequence was 100% identical to that of a Taiwanese isolate. The deduced amino acid sequences were 99 to 100% identical to those from other countries. In VP19, C and T in the foreign isolates were replaced by T and A in the Korean isolate at Positions 57 and 218 nt, respectively, downstream of A (+) of the VP19 start codon. The change at Position 218 nt resulted in valine in the foreign isolates being replaced by aspartate in the Korean isolate. PMID- 15352528 TI - Communicating effectively and managing care. PMID- 15352529 TI - Evaluating patient compliance with nurse advice line recommendations and the impact on healthcare costs. AB - OBJECTIVE: To explore the effect of telephone triage and advice lines in uninsured and managed care populations served by a safety net system and to document the relationship between the patient's initial plan for healthcare, the nurse recommendation, and the patient's subsequent healthcare action. STUDY DESIGN: Prospective telephone survey. PATIENTS AND METHODS: Of 1538 calls to a nurse advice line in a 28-day period, 710 (46%) callers were selected to be surveyed. Of those, 278 (39%) were surveyed by telephone within 7 days of their call to assess patient compliance with recommendations, the patient's actual healthcare actions, and their satisfaction with the service. RESULTS: Patients' reported actions were classified as either (1) home care (46%), (2) clinic visit (27%), or (3) hospital visit (27%). Seventy percent of patients complied with nurse advice line recommendations. Most patient actions (68%) differed from their original healthcare plan, with many (46%) choosing a lower intensity of care. Changes from patients' original healthcare plans had a potential annual net savings of $322 249. CONCLUSION: The simple act of calling a nurse triage and advice line corresponds with a change in the reported actions of uninsured and managed care patients and a potential reduction in costs to the safety net system providing their healthcare. PMID- 15352530 TI - Inflammatory bowel disease. AB - Inflammatory bowel disease is a complicated condition, including Crohn's disease, ulcerative colitis, microscopic colitis, and indeterminate colitis, that affects the intestine and several extraintestinal sites. There has been much debate regarding whether Crohn's disease and ulcerative colitis are distinct entities or if they exist along a continuum of the same disease process. In this article, the pathogenic mechanisms and clinical manifestations of inflammatory bowel disease are reviewed, as well as treatment options. Because Crohn's disease and ulcerative colitis are chronic diseases, they have an important economic effect on our healthcare system and the United States as a whole. Some newer and more expensive treatment options may provide overall cost savings in select patient populations because of decreased use of healthcare resources. PMID- 15352531 TI - Interventions to improve quality of care: the Kaiser Permanente-alzheimer's Association Dementia Care Project. AB - OBJECTIVE: To improve quality of dementia care in a Kaiser Permanente service area through rigorous dissemination of practice guidelines and social worker support for physicians and patients. STUDY DESIGN: Pre-post design with practice behavior change assessed by medical record review, and provider and caregiver satisfaction with care assessed by surveys. METHODS: A diagnostic guideline and later a management guideline were adopted for use by Kaiser Permanente physicians in metropolitan Los Angeles. Physicians received training based on the guidelines, and social workers provided ancillary support. Eighty-three community dwelling dementia patients and their caregivers were referred to the project by primary care physicians and then were assessed and followed by social workers. Data were abstracted from medical records to determine whether these interventions led to improved quality of care as indicated by adherence to key care processes derived from the adopted dementia guidelines. Chi-square and t tests were applied to compare guideline adherence and satisfaction rates before and after the interventions. RESULTS: Compared with baseline, higher rates of provider and caregiver satisfaction with Kaiser's system of dementia care were found at the postintervention follow-up. There also were significantly higher rates of adherence to several practice guideline-based quality measures: assessment of cognitive status; referrals to the Alzheimer's Association; and assessments of activities of daily living, decision-making capacity, depression, and wandering risk. CONCLUSION: Quality of primary care for people with dementia can be improved through guideline implementation with care management support by social workers. PMID- 15352532 TI - Associations with reduced length of stay and costs on an academic hospitalist service. AB - BACKGROUND: Prior studies of hospitalist services have suggested improved efficiency and quality of care compared with traditional inpatient services. OBJECTIVES: To compare outcomes of patients on a new hospitalist service with those on traditional inpatient services and to determine the impact of hospitalists on particular patient subgroups. STUDY DESIGN: Prospective, quasiexperimental, observational. METHODS: The study was conducted on the general medicine service at an academic teaching hospital, staffed by hospitalist physicians (HP) and nonhospitalist physicians (NHP), and included 1706 consecutive, directly admitted patients over 1 year. RESULTS: The 447 HP patients and 1259 NHP patients had similar rates of in-hospital mortality (1.3% vs 2.1%, respectively; P = .29) and 30-day readmission (7.8% vs 8.7%, respectively; P= .55). Mean hospital length of stay (LOS) was 1 day shorter for HP patients in unadjusted analyses (5.5 vs 6.5 days, respectively; P = .009) and in multivariable analyses adjusting for clustering and patient factors. Physician experience was not correlated (P < .2) with LOS. In stratified analyses, differences in LOS between HP and NHP patients were greater for patients residing closer to the hospital. Mean total costs were $917 less for HP patients (P = .08) and 10% less (P= .04) in multivariable analyses. Decreases in costs were significant (P < .05) for nursing ($604; P = .002) and laboratory services ($126; P = .04). Nonetheless, mean costs per day were $122 higher (P= .003) for HP patients. CONCLUSIONS: Patients managed by hospitalists had shorter LOS and lower costs than patients managed by nonhospitalists, but had higher costs per day. These results suggest that hospitalists increase the intensity of care and may have their greatest impact on specific types of patients and classes of hospital costs. PMID- 15352533 TI - CURATA: A patient health management program for the treatment of osteoarthritis in Quebec: an integrated approach to improving the appropriate utilization of anti-inflammatory/analgesic medications. AB - OBJECTIVES: To identify gaps in current osteoarthritis (OA) care in Quebec, Canada, and to implement and evaluate interventions to promote appropriate use of evidence-based medicine. STUDY DESIGN: Pretest and posttest; analysis of the Quebec health insurance database. METHODS: CURATA is a patient health management program utilizing an evidence-based approach for OA treatment. Evaluation of the current level of care revealed major gaps in physicians' knowledge of (1) risk factors for gastrointestinal (GI) toxicity associated with nonsteroidal anti inflammatory drugs (NSAIDs); (2) NSAID-induced toxicity associated with long-term administration and contraindications for NSAID use in patients with hypertension, cardiovascular disease, or renal insufficiency; (3) choice of cytoprotection; and (4) use of nonpharmacologic treatments for OA. The CURATA intervention consisted of educational workshops, with and without presentation of a decision tree regarding appropriate use of pharmacologic and nonpharmacologic OA treatments. Participating physicians were asked to complete an 8-item questionnaire before and after the workshop, as well as 3 and 6 months later, to test their immediate and remote knowledge of treatment choices. The prescribing patterns of GPs also were evaluated through analysis of the Quebec health insurance database. RESULTS: The participating physicians were better immediate and remote risk assessors of GI bleeding and made more appropriate treatment choices (15.2% improvement relative to mean preworkshop score). CONCLUSION: These evidence-based interventions were successful not only in improving the physicians' knowledge regarding the diagnosis and management of OA, but also--more importantly--in changing their behavior to make more appropriate therapy choices for their patients. PMID- 15352534 TI - New approaches to the management and treatment of attention-deficit/hyperactivity disorder. PMID- 15352535 TI - Innovations in attention-deficit/hyperactivity disorder pharmacotherapy: long acting stimulant and nonstimulant treatments. AB - This article reviews innovations in attentiondeficit/hyperactivity disorder (ADHD) pharmacotherapy and describes research on the newer, long-acting stimulant and nonstimulant treatments for ADHD. Results from peer-reviewed articles comparing the efficacy and safety of longer-acting methylphenidate or amphetamine based stimulants and the nonstimulant atomoxetine are described. Longer-acting stimulants and nonstimulants provide increased clinical utility compared with short-acting stimulants. Efficacy and safety are similar to 2- or 3-times-a-day treatment with short-acting stimulants. Longer-acting stimulants and nonstimulants provide increased convenience and flexibility for treating youth with ADHD and show considerable promise. Direct head-to-head studies are needed to better inform clinical decision making and to identify moderators and mediators of differential response. PMID- 15352536 TI - Attention-deficit/hyperactivity disorder: medication treatment-dosing and duration of action. AB - Several trends have affected the current standards for medication dosing recommendations for the treatment of attention-deficit/hyperactivity disorder (ADHD). Empirical evidence that treatment can effect improvement across multiple domains of impairment has encouraged optimizing treatment throughout the day. For stimulants, the value of increasing the dose to improve effect has replaced former notions of minimizing exposure to medication by treating at the lowest dose demonstrating effect. Several new stimulant formulations control the rate of dose delivery to optimize effect for longer intervals than immediate-release medication. This avoids morbidity associated with improper dose sculpting, inherent ups and downs of effect, and poorer adherence to multiple daily doses of shorter-acting products. Dosing of the new nonstimulant agent atomoxetine (Strattera) is based primarily on weight. Atomoxetine appears to have a longer duration of action in children and adolescents, but in adults it requires twice daily dosing to maintain effect across the day. Whereas its overall effect size is smaller than stimulant agents, there is no way to predict an individual's response in advance, and for some atomoxetine may be the optimal medication treatment. Some head-to-head comparison studies between long-acting stimulant preparations and atomoxetine, especially those focused on duration of effective action, are likely. PMID- 15352537 TI - Satisfaction with treatment for attention-deficit/hyperactivity disorder. AB - Patient satisfaction with treatment services is an important variable in ascertaining overall outcome; however, it is not a substitute for improving target symptoms and function. This article reviews the general determinants of satisfaction, including the effectiveness of the treatment, patient expectations, acceptability of the specific treatment, and provider factors as they apply to the treatment of attention-deficit/hyperactivity disorder (ADHD). Generally, parents and teachers are more receptive to or prefer nonmedication or behavioral therapies to medication, despite the established effectiveness of medication. Children are similarly ambivalent about medication. These preferences probably result from the lower acceptability of medication treatment for ADHD. Data does not establish greater patient satisfaction with a particular medication class or formulation. However, parents and children/adolescents may prefer the newer longer-acting medications. Measuring satisfaction with ADHD treatment serves several purposes, such as identifying acceptable treatment modalities, defining patient expectations for treatment, and determining those providers that may be deficient in communicating with their clients. PMID- 15352538 TI - New options in the pharmacological management of attention-deficit/hyperactivity disorder. AB - A brief review of the efficacy and common side effects of pharmacological treatments for attention-deficit/hyperactivity disorder (ADHD) is presented. Methylphenidate and amphetamine continue to have the strongest base of empiric support. They are both available in extended- and immediate-release preparations. Although most children with ADHD respond favorably to either medication, many children who do not respond to one medication will respond to the other. Additional pharmacological options include the nonstimulant atomoxetine, the stimulant pemoline, the antidepressants bupropion and desipramine, and the antihypertensives clonidine and guanfacine. In selecting the appropriate pharmacological regimen, consideration should be given to the child's daily school and after-school schedule, the presence of aggressive symptoms, and the risk of diversion. Careful selection of an appropriate medication regimen and active engagement of the child, parents, and teacher in daily management may help to ensure long-term adherence. PMID- 15352539 TI - Development and analytical validation of an enzyme linked immunosorbent assay for the measurement of canine gastric lipase immunoreactivity in serum. AB - The objective of this study was to develop and analytically validate an enzyme linked immunosorbent assay (ELISA) for measurement of canine gastric lipase immunoreactivity (cGLI). A sandwich ELISA was developed using canine gastric lipase (cGL) purified from canine stomachs and polyclonal antibodies directed against cGL, raised in rabbits and purified by affinity chromatography. The assay was validated by determination of sensitivity, working range, linearity, accuracy, precision, reproducibility, and the upper limit of the control range by determining the 97.5th percentile of serum cGLI concentration in 74 healthy canines. Sensitivity and working range in serum were 200 ng/L and 200 to 39 160 ng/L, respectively. Observed to expected ratios for dilutional parallelism for 3 serum samples and 3 dilutions ranged from 86.1% to 244.2% (mean +/- standard deviation [s]; 125.4% +/- 48.2%). Observed to expected ratios for spiking recoveries for 3 serum samples and 6 spiking concentrations ranged from 66.4% to 152.5% (mean +/- s; 104.5% +/- 22.9%). Intra-assay and interassay variabilities for 3 different serum samples were 25.5%, 9.4%, and 13.4% and 26.0%, 17.2%, and 14.4%, respectively. The upper limit of the control range for serum cGLI was 662 ng/L. We concluded that the ELISA for cGLI described here is highly sensitive and shows a wide working range. However, the validation characteristics for this assay are suboptimal and below values of approximately 2.000 ng/L the assay is more semiquantitative in nature. Despite its limitations, whether this assay is useful for the diagnosis of canine gastric disorders remains to be determined. PMID- 15352540 TI - Bulk tank milk urea nitrogen: seasonal patterns and relationship to individual cow milk urea nitrogen values. AB - The objectives of this study were: 1) to determine if bulk tank milk urea nitrogen (BTMUN) and whole herd weighted average of the individual cow MUN levels (WHMUN) were equivalent measurements of herd MUN status; and 2) to determine the seasonal variation in BTMUN concentrations in Prince Edward Island (PEI) dairy herds. For BTMUN-WHMUN correlation testing, bulk tank milk samples from 176 herds were tested for MUN once every 1 to 2 wk between September 1999 and August 2002, as part of routine BTM testing for milk components. During this 3-year period, all herds had all milking cows tested for MUN once a month at the same lab. The WHMUN levels (weighted for milk production) were calculated for each month, and were compared to BTMUN levels using a concordance correlation coefficient (CCC) and a graphic procedure. Tests were only compared if they occurred on the same date, producing a final dataset of 669 comparisons. The BTMUN had good (but not perfect) correlation with WHMUN (CCC = 0.91). This high reliability extended to both the pasture and non-pasture seasons, various milk sampling protocols, and all herd sizes seen in PEI. For evaluating the seasonal variation of BTMUN, the 3 y worth of data (24 803 observations) were divided into 15 seasonal categories, 5 seasons per year (early, mid, and late pasture, and early and late stable). Using linear mixed modelling, significantly (P < 0.05) higher BTMUN values were found during the mid and late pasture seasons of 2000, likely because the precipitation was unusually high during this period, enhancing pasture growth. PMID- 15352541 TI - Susceptibility of Mycobacterium avium sbsp paratuberculosis to monensin sodium or tilmicosin phosphate in vitro and resulting infectivity in a murine model. AB - This study was designed to determine the susceptibility in vitro and infectivity of 1 field isolate of Mycobacterium avium sbsp paratuberculosis after exposure to monensin sodium and tilmicosin phosphate. Minimum inhibitory concentrations (0.39 microg monensin sodium/mL; 1.60 microg tilmicosin phosphate/mL) were determined in quintuplicate. Organisms were then incubated with 3 different concentrations of each medication for 3 different lengths of time, then washed and resuspended in sterile physiologic saline and injected intraperitoneally into mice that were genetically susceptible to infection. Mice were euthanatized 50 d later and the number of hepatic granulomas was used as the indicator of infectivity. Neither time of incubation nor concentration of medication had any effect on the infectivity of the organisms. Monensin sodium significantly reduced the number of hepatic granulomas in genetically susceptible mice while tilmicosin phosphate did not. Antimycobacterial activity of monensin sodium suggests that the role of monensin in the control of bovine paratuberculosis should be evaluated further. PMID- 15352542 TI - Effect of intramammary injection of rboGM-CSF on milk levels of chemiluminescence activity, somatic cell count, and Staphylococcus aureus count in Holstein cows with S. aureus subclinical mastitis. AB - The effect of intramammary injection of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF, 400 microg/10 mL) on quarter milk levels of chemiluminescence (CL) activity, and somatic cell count (SCC) and shedding pattern of Staphylococcus aureus was investigated. Ten Holstein cows, naturally infected with S. aureus were used, with either early-stage or late-stage subclinical mastitis. Injection of rboGM-CSF caused a remarkable increase in milk CL activity with a peak at 6 h after the cytokine injection in the early- and late-stage groups. In the early-stage group, milk SCC stayed around preinjection level at 6 h, rose significantly on days 1 and 2, and was followed by a smooth and significant decline to an under preinjection level (below 200 000 cells/mL) on day 7 postinjection. Alternatively, in the late-stage group, milk SCC rose significantly at 6 h after the cytokine injection and maintained high levels thereafter. The milk S. aureus count decreased drastically by the cytokine injection in the early-stage group. The bacterial count was moderately decreased in the late-stage group, but increased back to preinoculation levels on day 7 after the cytokine injection. The results suggest that the rboGM-CSF has a potential as a therapeutic agent for S. aureus infection causing subclinical mastitis of dairy cows, if the cytokine is applied at the initial stage of infection. PMID- 15352543 TI - Bacteriology and somatic cell counts in milk samples from ewes on a Scottish farm. AB - Milk samples from 50 sheep on a single Scottish research farm were collected weekly for 10 wk postpartum. Samples were analyzed for somatic cell counts (SCC) each week and bacteriologic culture was done for 7 of the 10 wk. A total of 492 udder half samples were cultured, of which 467 had corresponding cell count data. Statistical analysis on complete SCC and culture data showed no association between SCC and bacterial isolation, even when more than 10 colonies of a single bacterial species were present. Only 3.6% of the samples were simultaneously positive for high count (> 10 colonies from 0.01 mL of milk) of any one bacterial species and high SCC (> 1 x 10(6)/mL). The bacteria recovered were: Staphylococcus equorum (19 times), S. xylosus (7 times), S. simulans (6 times), Streptococcus uberis (3 times) and other streptococci (4 times), Mannheimia (Pasteurella) haemolytica (2 times), Staphylococcus aureus (1 time), S. capitis (1 time), and Enterococcus faecium (1 time). There was an association between the test day and SCC, with higher SCC values in the first 2 wk. In addition, significantly higher SCC values were found in the oldest animals compared to the other age groups. PMID- 15352544 TI - Clinical signs and hematologic, cytokine, and plasma nitric oxide alterations in response to Strongylus vulgaris infection in helminth-naive ponies. AB - The objective of this study was to determine the effect of infection with Strongylus vulgaris on serum cytokines and plasma nitric oxide (NO) concentrations in helminth-naive ponies. Group 1 (n = 21) was given 500 S. vulgaris L3 larvae and group 2 (n = 7) received a saline control. Ponies were monitored daily for clinical signs, and blood was collected for complete blood cell counts and serum cytokines (TNF, IL-1, IL-6) quantification. Group 1 ponies were depressed, anorexic, and febrile for variable periods of time. Plasma NO was increased on day 21 in group 1 and on days 9 and 21 in group 2. Significant increases in total white blood cell counts, fibrinogen, and plasma protein concentrations in group 1 were found. Significant decreases in red blood cell counts and packed cell volume were also noted in group 1. There were no differences in serum cytokines across time in either group of ponies. Despite the lack of proinflammatory cytokine induction with the apparent inflammatory response to S. vulgaris there is evidence of a potential role of NO. PMID- 15352545 TI - Differences in expression of retinal pigment epithelium mRNA between normal canines. AB - A reference database of differences in mRNA expression in normal healthy canine retinal pigment epithelium (RPE) has been established. This database identifies non-informative differences in mRNA expression that can be used in screening canine RPE for mutations associated with clinical effects on vision. Complementary DNA (cDNA) pools were prepared from mRNA harvested from RPE, amplified by PCR, and used in a subtractive hybridization protocol (representational differential analysis) to identify differences in RPE mRNA expression between canines. The effect of relatedness of the test canines on the frequency of occurrence of differences was evaluated by using 2 unrelated canines for comparison with 2 female sibling canines of blue heeler/bull terrier lineage. Differentially expressed cDNA species were cloned, sequenced, and identified by comparison to public database entries. The most frequently observed differentially expressed sequence from the unrelated canine comparison was cDNA with 21 base pairs (bp) identical to the human epithelial membrane protein 1 gene (present in 8 of 20 clones). Different clones from the same-sex sibling RPE contained repetitions of several short sequence motifs including the human epithelial membrane protein 1 (4 of 25 clones). Other prevalent differences between sibling RPE included sequences similar to a chicken genetic marker sequence motif (5 of 25), and 6 clones with homology to porcine major histocompatibility loci. In addition to identifying several repetitively occurring, noninformative, differentially expressed RPE mRNA species, the findings confirm that fewer differences occurred between siblings, highlighting the importance of using closely related subjects in representational difference analysis studies. PMID- 15352546 TI - An assessment of sanitation protocols for commercial transport vehicles contaminated with porcine reproductive and respiratory syndrome virus. AB - The objective of this study was to develop and test a rapid (< 2 h) sanitation protocol designed for porcine reproductive and respiratory syndrome virus (PRRSV) positive commercial transport vehicles involving cold water washing and disinfection via fumigation using scale models of weaned pig trailers. The study consisted of 2 phases. Following experimental contamination of model trailers with PRRSV MN 30-100 (5 x 10(5)TCID50), phase 1 evaluated the presence or absence of PRRSV RNA by polymerase chain reaction (PCR) on swabs collected from the trailer interiors 0, 60, and 90 min after treatment. Phase 2 consisted of evaluating the infectivity of trailers 90 min posttreatment by monitoring changes in the PRRSV-status of naive sentinel pigs housed for 2 h. Treatments included washing only (treatment 1), washing plus formaldehyde fumigation (treatment 2), washing plus fumigation with glutaraldehyde-quaternary ammonium chloride (treatment 3), and washing plus overnight drying (treatment 4). Porcine reproductive and respiratory syndrome virus RNA was detected in all trailers (20 out of 20 replicates) at 60 and 90 min following the application of treatments 1 and 2. These trailers also contained infectious PRRSV, as determined by the infection of naive pigs housed in treated trailers and the testing of organic debris collected from the interior of trailers by swine bioassay. At 90 min posttreatment, all trailers treated with glutaraldehyde-quaternary ammonium chloride were PCR-negative, non-infectious to sentinel pigs, and swine bioassay negative. Similar results were observed in trailers allowed to dry for 8 h. Under the conditions of this study, it appears certain disinfectants may possess different levels of efficacy against PRRSV and PRRSV-positive models may be effectively sanitized in the absence of overnight drying. PMID- 15352547 TI - Exposure of sero-positive gilts to swine influenza virus may cause a few stillbirths per litter. AB - Six pregnant gilts were purchased from a high health herd and were found to be serologically positive for swine influenza virus (SIV) subtype H3N2. Three of the gilts, at 80 to 82 days of gestation, were experimentally exposed a second time to the same SIV subtype--H3N2. No clinical signs resulted from the second exposure to SIV and hemagglutination-inhibition (HI) titers for SIV at 4 weeks postexposure were unchanged suggesting that the gilts had not been reinfected. However, the second exposure to SIV affected the number of pigs born alive. Each of the 3 litters from the twice exposed gilts suffered 2 or 3 stillborn piglets per litter. In contrast the 3 matched, sero-positive gilts that were not exposed to SIV (controls) had no stillborn piglets. These differences were statistically significant using a t-test for unequal variances (P = 0.0086). Sera from 2 of the stillborn piglets were negative for HI antibodies and there was no indication from the pigs born alive that the H3N2 virus had crossed the placenta. PMID- 15352548 TI - Enteritis associated with porcine circovirus 2 in pigs. AB - This report describes the diagnosis of porcine circovirus 2 (PCV2)-associated enteritis in 6 weaned pigs without postweaning multisystemic wasting syndrome by histopathology, virus isolation, and in situ hybridization. The most unique lesions were granulomatous inflammation affecting Peyer's patches, characterized by infiltrates of epithelioid macrophages and giant multinucleated cells. Large, multiple, basophilic or amphophilic, grape-like intracytoplasmic inclusion bodies were often seen in the cytoplasm of histiocytic cells and giant multinucleated cells. No microscopic lesions were observed in the lymphoid tissue, such as lymph node, spleen, and tonsil. A strong hybridization signal for PCV2 was detected in the cytoplasm of histiocytes and giant multinucleated cells in Peyer's patches. Porcine circovirus 2 was isolated from homogenates of the small and large intestines in 2 weaned pigs. The presence of diarrhea and granulomatous enteritis, and abundant PCV2 DNA associated with the microscopic lesions is suggestive of PCV2-associated enteritis. Thus, PCV2-associated enteritis could be a distinct clinical manifestation of PCV2. PMID- 15352549 TI - An attempt to render oocytes and embryos free from the porcine circovirus type 2 after experimental in vitro exposure. AB - The purpose of this study was to investigate the possibility of rendering oocytes and embryos free of porcine circovirus type 2 (PCV2). Groups of cumulus oocytes complexes, cumulus free oocytes, and embryos 3 to 5 d post breeding were exposed to PCV2 (10(5) TCID50/mL) prior to disinfection by washing and different combinations of enzymatic treatments. The study suggests that under the in vitro conditions used, standard washing procedures with, or without, trypsin or incorporating pronase or hyaluronidase and DNase/RNase in the treatment was not effective in rendering oocytes and embryos free from PCV2 nucleic acid. Since the virus is noncytopathic in cell culture and for embryonic cells, it appears that there is a possibility of introducing viral contamination through oocytes collected from infected pigs into the in vitro fertilization system with subsequent potential of producing in vitro fertilized embryos associated with PCV2. PMID- 15352550 TI - Combined vaccination of live 1B Chlamydophila abortus and killed phase I Coxiella burnetii vaccine does not destroy protection against chlamydiosis in a mouse model. AB - Q fever and chlamydiosis often affect ovine and caprine flocks simultaneously or successively. Combination vaccines effective against these 2 diseases would be of great value in veterinary medicine. Unfortunately, the current effective vaccines are a live vaccine for chlamydiosis and killed vaccine for Q fever. Vaccination of mice with live chlamydiosis vaccine 1B and killed phase I vaccine against Q fever at 2 points on the back at the same time produced good protection against chlamydial abortion. This suggests that it may be possible to vaccinate ewes and goats against chlamydiosis and Q fever simultaneously. PMID- 15352551 TI - Molecular basis of quinolone resistance in Escherichia coli from wild birds. AB - Nine quinolone resistant (minimal inhibitory concentration [MIC] was > 32 microg/mL for nalidixic acid, > 1 microg/mL for ciprofloxacin) isolates of Escherichia coli have been found in wild birds with septicemia. All of the isolates were aerobactin positive. The mechanisms of resistance were characterised by sequencing the quinolone resistance-determining region (QRDR) of the gyrA, gyrB, parC, and parE genes. Sequence analysis of the gyrA gene in all isolates identified only 1 nucleotide substitution at codon Serine-83 for Leucine 83. Sequence analysis of the gyrB, parC, and parE QRDR genes revealed no mutations in any of the isolates. This study was conducted to determine the importance of these genes in the susceptibility of E. coli strains isolated from wild birds to quinolones. PMID- 15352552 TI - In vitro regulation of Mac-1 expression on bovine polymorphonuclear leukocytes by endotoxin and tumor necrosis factor-alpha at different stages of lactation. AB - The purpose of this in vitro study is to clarify some of the underlying mechanisms leading to the decreased migratory capacity of polymorphonuclear leukocytes (PMN) during mastitis in dairy cows soon after calving. Surface expression of Mac-1 (CD11b, CR3) on PMN and of CD14 on monocytes was measured in early- (EL), peak- (PL), and midlactation (ML) by flow cytometric analysis. In addition, we evaluated the effect of lipopolysaccharide (LPS) and tumor necrosis factor (TNF)-alpha on CD11b surface expression in PMN at different stages of lactation in a whole blood model. During EL, while resting monocytes expressed diminished levels of CD14, the basal expression of CD11b on PMN was not significantly altered. The relative increase of CD11b on PMN after incubation with LPS or TNF-alpha did not significantly differ among EL, PL, or ML at any of the concentrations tested. The current findings do not support an important role for basal CD11b levels nor for a defective mobilization of CD11b by LPS and TNF alpha in the reduced migratory capacity of PMN during EL. PMID- 15352554 TI - From X-ray technician to radiologic technologist. PMID- 15352553 TI - Evaluation of a ceftiofur-washed whole cell Streptococcus suis bacterin in pigs. AB - The efficacy of currently available washed whole cell Streptococcus suis bacterins is generally poor. We developed and tested the efficacy of a novel ceftiofur-washed whole cell bacterin. Sixty-six, 2-week-old specific pathogen free (SPF) pigs were randomly divided into 5 groups. Three groups were vaccinated 28 and 14 d prior to challenge. The 3 ceftiofur-washed whole cell bacterins each contained 1 of 3 different adjuvants (Montanide ISA 25, Montanide ISA 50, and Saponin). Pigs exhibiting severe central nervous system disease or severe joint swelling and lameness were euthanized immediately and necropsied. All remaining pigs were necropsied at 14 d post inoculation. The ceftiofur-washed whole cell S. suis bacterin with Montanide ISA 50 adjuvant significantly (P < 0.05) reduced bacteremia, meningitis, pneumonia, and mortality associated with S. suis challenge. Further work on this novel approach to bacterin production is warranted. PMID- 15352555 TI - Human oocyte radiosensitivity. AB - The teratogenic effects of ionizing radiation are well documented, but less is known about the radiosensitivity of the human oocyte. Women of childbearing age are understandably concerned about diagnostic radiology examinations--their risk/benefit ratio and their impact on the ability to conceive. Current research and evidence gathered from studies of the long-term effects of intra-abdominal therapeutic radiation shed light on this subject. With respect to this issue, radiographers can now give more practical advice to their nongravid female patients. PMID- 15352556 TI - Measuring mammography patient satisfaction. AB - This study identified how the mammography industry addresses patient needs and satisfaction through consumer satisfaction mechanisms. Survey results from 500 randomly selected mammography facilities show that more than 50% of the facilities make an effort to address patient satisfaction. Results highlight the need for institutions to critically review reliability and validity criteria of satisfaction instruments, thus enabling valuable organizational change. PMID- 15352557 TI - Digital image processing. AB - Digital image processing is now commonplace in radiology, nuclear medicine and sonography. This article outlines underlying principles and concepts of digital image processing. After completing this article, readers should be able to: List the limitations of film-based imaging. Identify major components of a digital imaging system. Describe the history and application areas of digital image processing. Discuss image representation and the fundamentals of digital image processing. Outline digital image processing techniques and processing operations used in selected imaging modalities. Explain the basic concepts and visualization tools used in 3-D and virtual reality imaging. Recognize medical imaging informatics as a new area of specialization for radiologic technologists. PMID- 15352558 TI - Self-leadership: becoming an exceptional leader. AB - This article describes the importance of self-leadership for health care workers. By continually evaluating and improving leadership skills, health care workers and organizations can develop a work environment that encourages individual growth and trains leaders for future success. After completing this article, the reader will be able to: Understand the impact values and emotions have on daily interaction. Recognize personal strengths and weaknesses through self-evaluation. Understand how emotional and personality profile tests aid in the self-discovery process. Explain the importance of physical and mental well-being. Understand the value of establishing a life-long leadership plan. PMID- 15352559 TI - Remedying an educator shortage. PMID- 15352560 TI - Taking the time to talk. PMID- 15352561 TI - A computed radiography challenge. PMID- 15352562 TI - Student supervision and patient care. PMID- 15352563 TI - Safety alert for transdermal patches. PMID- 15352564 TI - PET helps uncover undiagnosed neurological syndromes. PMID- 15352565 TI - Thoracic PET appropriate for diagnosing some lung cancers. PMID- 15352566 TI - MDCT angiography may help identify patients in danger of stroke. PMID- 15352567 TI - Patient page. Lower extremity radiography. PMID- 15352568 TI - The looming threat of HIV/AIDS drug resistance in South Africa. PMID- 15352569 TI - Screening for breast cancer. PMID- 15352570 TI - Screening for breast cancer. PMID- 15352571 TI - Screening for breast cancer. PMID- 15352572 TI - Screening for breast cancer. PMID- 15352573 TI - Failed contraception? PMID- 15352574 TI - Medical practitioners' attitudes towards older patients. PMID- 15352575 TI - ARV compliance--the new battle terrain. PMID- 15352576 TI - Drug laws--state to comply... but later. PMID- 15352577 TI - Winning back mother's magical milk. PMID- 15352578 TI - Cancer sufferers could postpone pregnancy. PMID- 15352579 TI - Marketing your medical practice--Part II. PMID- 15352580 TI - Prescribed minimum benefits or minimum prescribed benefits? PMID- 15352581 TI - Sedation of children undergoing MRI--a risky business! PMID- 15352582 TI - Occupational post-exposure HIV prophylaxis. PMID- 15352583 TI - Usefulness of neuro-imaging for the diagnosis of HIV encephalopathy in children. PMID- 15352584 TI - Universal newborn hearing screening in South Africa--a First-World dream? PMID- 15352585 TI - I want my Mirena out! PMID- 15352586 TI - Gender distribution of students and staff at the University of Malawi College of Medicine, 1991-2003. PMID- 15352587 TI - Access to health care in South Africa--the influence of race and class. AB - OBJECTIVES: The first democratic government elected in South Africa in 1994 inherited huge inequities in health status and health provision across all sections of the population. This study set out to assess the impact of the new government's commitment to address these inequities and implement policies to improve population health in general and address inequalities in health care in particular. DESIGN: A 1998 household survey assessed many aspects of health delivery, including their own perceived and actual access to health care among different segments of South African society. RESULTS: Race was the main predictor of perceived changes in access to health care, with black, coloured-and Indian respondents significantly more likely to feel that access had improved since 1994, compared with white respondents. Socio-economic status (SES) was the main predictor of actual access to health care, with low and middle SES classes significantly less likely to access care when ill. CONCLUSIONS: One-third of respondents perceived health care access to have improved between 1994 and 1998, and this response was partially determined along racial lines. About one-quarter reported an inability to access health care when they required it, and this response was partially determined along socio-economic lines. This set of contrasting responses suggests that at a political level perceptions are largely influenced by race, but at the operational level actual access is influenced by SES. PMID- 15352588 TI - Initial experience of a public sector antiretroviral treatment programme for HIV infected children and their infected parents. AB - OBJECTIVE: To describe the initial experience of treating HIV-infected children and their infected parents with antiretroviral therapy. DESIGN: Prospective, cohort study. SETTING: Tertiary, referral hospital. PATIENTS: HIV-infected children and their parents. METHODS: This report focuses on the early response of children to highly active antiretroviral therapy (HAART). Children were followed up at 4-weekly intervals. Monitoring included initial and yearly viral load measurements, baseline and 6-monthly CD4 counts and 4-weekly adherence checks. RESULTS: Between August 2002 and June 2003, 80 children were enrolled in the programme, representing a follow-up period of 23.9 patient-years. Seventy-five children had severe clinical disease, severe immune suppression, or a combination of the two. The response of children who had received HAART for > or = 6 months (N = 17) was assessed. There was no change in mass z-score (p = 0.11) or length z score (p = 0.37), but a significant increase in CD4 percentage (p < 0.0001) during the first 6 months of therapy. Six-month viral loads were available for 12 children. There was a significant drop in viral load (p = 0.001) and 9 achieved undetectable levels by 6 months. Most children achieved > or = 85% adherence. By June 2002, 67 children (84%) were relatively well, 1 had B-cell lymphoma, 7 (8.8%) had died, 4 (5%) were lost to follow-up and 1 was withdrawn from the programme. Of 57 children who completed 3 months of HAART, 12 were admitted a total of 17 times for infectious complications. There were no severe drug reactions. Three of 7 mothers on HAART received treatment through the programme. CONCLUSION: These initial results suggest that many HIV-infected children in the public sector will benefit from antiretroviral therapy. However, both ambulatory and inpatient facilities are required to manage children on HAART comprehensively. PMID- 15352589 TI - Imiglucerase low-dose therapy for paediatric Gaucher disease--a long-term cohort study. AB - BACKGROUND: Gaucher disease is the most common lysosomal storage disorder caused by the insufficiency of the lysosomal enzyme, glucocerebrosidase. This deficiency results in absent or inefficient conversion of glucocerebroside (a membrane lipid) to ceramide and glucose. Accumulation of glucocerebroside occurs primarily in macrophage lysosomes (i.e. monocytes and macrophages) during phagocytic degradation of red blood cells. Clinical symptoms arise due to the displacement of normal cells by lipid-engorged Gaucher cells. Enzyme replacement therapy (ERT) targets the macrophage system and has been shown to be successful in the treatment of type 1 Gaucher disease in adults and children. ERT (60 U/kg) every 2 weeks decreases and often reverses organomegaly and haematological complications and improves quality of life for patients with type 1 Gaucher disease. The present study describes the course of 9 paediatric patients followed up for 2 - 10 years receiving low-dose imiglucerase therapy (+/- 10 U/kg every 2 weeks) for moderate to severe type 1 Gaucher disease. OBJECTIVES: To evaluate the efficacy of low-dose imiglucerase therapy in paediatric Gaucher disease. SUBJECTS AND METHODS: Data were recorded at a single centre for 9 paediatric patients. Assessment of response included serial measurements of haemoglobin (Hb) concentrations, platelet count, angiotensin-converting enzyme (ACE) and total acid phosphatase (TAP) levels. Growth was assessed by serial determinations of body weight and height, plotted against standard growth charts. Organ size (liver and spleen) was measured clinically and also radiologically, where possible. RESULTS: In this low-dose imiglucerase treatment group: (i) there was a significant increase in Hb over time--normal Hb levels were achieved in 7 of the 9 patients after a mean of 3.7 years; (ii) platelet counts increased over time, reaching normal levels in 7 patients; (iii) there was a significant decrease in both ACE and TAP over time; (iv) heights and weights of the subjects increased significantly over time with treatment, normalising to the expected growth percentiles; and (v) organ size (liver and spleen) reduced with therapy in all patients measured. CONCLUSION: ERT with low-dose imiglucerase (+/- 10 U/kg/fortnight) ameliorates Gaucher disease-associated anaemia and thrombocytopenia. Low-dose ERT is effective and may be considered in resource poor clinical situations when other alternatives are not available. PMID- 15352590 TI - The role of the traditional bonesetter in primary fracture care in Nigeria. AB - BACKGROUND: In view of the growing interest in the management of fracture worldwide, the traditional bonesetter (TBS) practice in Nigeria was documented. OBJECTIVE: To highlight the role of the TBS in primary fracture care in Nigeria. SETTING: Rural. STUDY DESIGN: Over a 5-year period a qualitative study of the TBS settings and knowledge, diagnostic techniques and principles of fracture treatment in four TBS centres in Nigeria was carried out by active participation, on the spot assessment, interactive dialogues and oral interviews. RESULTS: It was found that TBS services are well preserved as a family practice, and training is by apprenticeship. Records are kept by oral tradition. There is no prescribed fee and the patronage is high. Fracture diagnosis is based on physical assessment and experience. The TBS relies solely on the conservative method of fracture treatment, and all fractures are reduced by the closed method and stabilised with an external traditional splint and a protracted period of immobilisation. The outcome of TBS treatment is good for closed fractures of the shaft of the humerus, ulna, radius and tibia, but poor for peri-articular and open fractures. Non-union, malunion, traumatic osteomyelitis and limb gangrene were the common major complications of TBS treatment. CONCLUSION: Despite criticisms and antagonism from orthodox medical practitioners TBS practice is well patronised by Nigerians. In order to guarantee safety and efficiency of the TBS practice in primary fracture care service delivery in Nigeria, there is therefore a need both to educate the community and to train the TBS. PMID- 15352591 TI - Surgical repair of brachial plexus injury. PMID- 15352592 TI - Surgical repair of brachial plexus injury: a multinational survey of experienced peripheral nerve surgeons. AB - OBJECT: Brachial plexus injuries (BPIs) are often devastating events that lead to upper-extremity paralysis, rendering the limb a painful extraneous appendage. Fortunately, there are several nerve repair techniques that provide restoration of some function. Although there is general agreement in the medical community concerning which patients may benefit from surgical intervention, the actual repair technique for a given lesion is less clear. The authors sought to identify and better define areas of agreement and disagreement among experienced peripheral nerve surgeons as to the management of BPIs. METHODS: The authors developed a detailed survey in two parts: one part addressing general issues related to BPI and the other presenting four clinical cases. The survey was mailed to 126 experienced peripheral nerve physicians and 49 (39%) participated in the study. The respondents represent 22 different countries and multiple surgical subspecialties. They performed a mean of 33 brachial plexus reconstructions annually. Areas of significant disagreement included the timing and indications for surgical intervention in birth-related palsy, treatment of neuroma-in-continuity, the best transfers to achieve elbow flexion and shoulder abduction, the use of intra- or extraplexal donors for motor neurotization, and the use of distal or proximal coaptation during nerve transfer. CONCLUSIONS: Experienced peripheral nerve surgeons disagree in important ways as to the management of BPI. The decisions made by the various treating physicians underscore the many areas of disagreement regarding the treatment of BPI, including the diagnostic approach to defining the injury, timing of and indications for surgical intervention in birth-related palsy, the treatment of neuroma-in-continuity, the choice of nerve transfers to achieve elbow flexion and shoulder abduction, the use of intra- or extraplexal donors for neurotization, and the use of distal or proximal coaptation during nerve transfer. PMID- 15352593 TI - Evaluation of suprascapular nerve neurotization after nerve graft or transfer in the treatment of brachial plexus traction lesions. AB - OBJECT: The aim of this retrospective study was to evaluate the restoration of shoulder function by means of suprascapular nerve neurotization in adult patients with proximal C-5 and C-6 lesions due to a severe brachial plexus traction injury. The primary goal of brachial plexus reconstructive surgery was to restore biceps muscle function and, secondarily, to reanimate shoulder function. METHODS: Suprascapular nerve neurotization was performed by grafting the C-5 nerve in 24 patients and by accessory or hypoglossal nerve transfer in 29 patients. Additional neurotization involving the axillary nerve was performed in 18 patients. Postoperative needle electromyography studies of the supraspinatus, infraspinatus, and deltoid muscles showed signs of reinnervation in most patients; however, active glenohumeral shoulder function recovery was poor. In nine (17%) of 53 patients supraspinatus muscle strength was Medical Research Council (MRC) Grade 3 or 4 and in four patients (8%) infraspinatus muscle power was MRC Grade 3 or 4. In 18 patients in whom deltoid muscle reinnervation was attempted, MRC Grade 3 or 4 function was demonstrated in two (11%). In the overall group, eight patients (15%) exhibited glenohumeral abduction with a mean of 44 +/- 17 degrees (standard deviation [SD]; median 45 degrees) and four patients (8%) exhibited glenohumeral exorotation with a mean of 48 +/- 24 degrees (SD; median 53 degrees). In only three patients (6%) were both functions regained. CONCLUSIONS: The reanimation of shoulder function in patients with proximal C-5 and C-6 brachial plexus traction injuries following suprascapular nerve neurotization is disappointingly low. PMID- 15352594 TI - Radiosurgery for arteriovenous malformations. PMID- 15352595 TI - Do the morphological characteristics of arteriovenous malformations affect the results of radiosurgery? AB - OBJECT: The authors sought to determine which morphological features of arteriovenous malformations (AVMs) are statistically predictive of preradiosurgical hemorrhage, postradiosurgical hemorrhage, and neuroimaging defined failure of radiosurgical treatment. In addition, correlation between computerized tomography (CT) scanning and angiography for the identification of AVM structures was investigated. METHODS: Archived CT dosimetry and available angiographic and clinical data for 268 patients in whom AVMs were treated with linear accelerator radiosurgery were retrospectively reviewed. Many of the morphological features of AVMs, including location, volume, compact or diffuse nidus, neovascularity, ease of nidus identification, number of feeding arteries, location (deep or superficial) of feeding arteries, number of draining veins, deep or superficial venous drainage, venous stenoses, venous ectasias, and the presence of intranidal aneurysms, were analyzed. In addition, a number of patient and treatment factors, including patient age, presenting symptoms, radiation dose, repeated treatment, and radiological outcome, were subjected to multivariate analyses. Two hundred twenty-seven patients were treated with radiosurgery for the first time and 41 patients underwent repeated radiosurgery. Eighty-one patients presented with a history of AVM hemorrhage and 91 patients had AVMs in a periventricular location. Twenty-six patients (10%) experienced a hemorrhage following radiosurgery. Of the 268 patients, 81 (30%) experienced angiographically defined cures, and 37 (14%) experienced MR imaging-defined cures. Eighty-six patients (32%) experienced neuroimaging-defined treatment failure, and 64 underwent insufficiently long follow up. A larger AVM volume (odds ratio [OR] 0.349; p = 0.004) was associated with a decreased rate of pretreatment hemorrhage, whereas periventricular location (OR 6.358; p = 0.000) was associated with an increased rate of pretreatment hemorrhage. None of the analyzed factors was predictive of hemorrhage following radiosurgery. A higher radiosurgical dose was strongly correlated with neuroimaging-defined success (OR 3.743; p = 0.006), whereas a diffuse nidus structure (OR 0.246; p = 0.008) and associated neovascularity (OR 0.428; p = 0.048) were each associated with a lower neuroimaging-defined cure rate. A strong correlation between CT scanning and angiography was noted for both nidus structure (p = 0.000; Fisher exact test) and neovascularity (p = 0.002; Fisher exact test). CONCLUSIONS: Patients presenting with AVMs that are small or periventricular were at higher risk for experiencing hemorrhage. A higher radiosurgical dose correlated strongly with neuroimaging defined success. Patients in whom the AVM had a diffuse structure or associated neovascularity were at higher risk for neuroimaging-defined failure of radiosurgery. A strong correlation between CT scanning and angiography in the assessment of AVM structure was demonstrated. PMID- 15352596 TI - Shunt-dependent hydrocephalus after rupture of intracranial aneurysms: a prospective study of the influence of treatment modality. AB - OBJECT: This study was designed to determine whether the frequency of shunt dependent hydrocephalus in patients suffering from aneurysmal subarachnoid hemorrhage (SAH) differs when comparing surgical clip application with endovascular obliteration of ruptured aneurysms. METHODS: In this prospective nonrandomized study, 245 patients with aneurysmal SAH treated using either surgical clip application or endovascular coil embolization were studied at our institution between September 1997 and March 2003. One hundred eighty patients underwent clip application and 65 had coil embolization. In those patients who underwent clip application of anterior circulation aneurysms, the lamina terminalis was systematically fenestrated. The occurrence of acute, asymptomatic, and shunt-dependent hydrocephalus was analyzed in both treatment groups. A subgroup analysis of patients with good clinical grade (World Federation of Neurosurgical Societies [WFNS] Grades I-III) and better Fisher Grade (1-3) and of patients with Fisher Grade 4 hemorrhage was performed. Acute hydrocephalus was observed in 19% of surgical cases and 46% of endovascular ones. The occurrence of asymptomatic hydrocephalus was similar in both treatment groups (p = 0.4). Shunt dependent hydrocephalus occurred in 14% of surgical cases and 19% of endovascular cases. This difference did not reach statistical significance (p = 0.53). Logistic regression models controlling for patient age, WFNS grade, Fisher grade, and acute hydrocephalus in patients with good clinical grade and better Fisher grade revealed no significant difference in the rate of shunt-dependent hydrocephalus in both therapy groups (odds ratio [OR] 0.8, 95% confidence interval [CI] 0.2-2.65). Results of similar models indicated that among patients with intraventricular hemorrhage (IVH), surgical clip application carried a lower risk of shunt-dependent hydrocephalus (OR 0.32, 95% CI 0.14-0.75) compared with that for endovascular embolization. CONCLUSIONS: Shunt-dependent hydrocephalus was comparable in the two treatment groups, even in patients with better clinical and radiological grades on admission. Only patients in the endovascular therapy group who had experienced IVH showed a higher likelihood of shunt-dependent hydrocephalus. PMID- 15352597 TI - Relationship between intracranial pressure and other clinical variables in patients with aneurysmal subarachnoid hemorrhage. AB - OBJECT: Increased intracranial pressure (ICP) is well known to affect adversely patients with head injury. In contrast, the variables associated with ICP following aneurysmal subarachnoid hemorrhage (SAH) and their impact on outcome have been less intensely studied. METHODS: In this retrospective study the authors reviewed a prospective observational database cataloging the treatment details in 433 patients with SAH who had undergone surgical occlusion of an aneurysm as well as ICP monitoring. All 433 patients underwent postoperative ICP monitoring, whereas only 146 (33.7%) underwent both pre- and postoperative ICP monitoring. The mean maximal ICP was 24.9 +/- 17.3 mm Hg (mean +/- standard deviation). During their hospital stay, 234 patients (54%) had elevated ICP (> 20 mm Hg), including 136 of those (48.7%) with a good clinical grade (Hunt and Hess Grades I-III) and 98 (63.6%) of the 154 patients with a poor grade (Hunt and Hess Grades IV and V) on admission. An increased mean maximal ICP was associated with several admission variables: worse Hunt and Hess clinical grade (p < 0.0001), a lower Glasgow Coma Scale (GSC) motor score (p < 0.0001); worse SAH grade based on results of computerized tomography studies (p < 0.0001); intracerebral hemorrhage (p = 0.024); severity of intraventricular hemorrhage (p < 0.0001); and rebleeding (p = 0.0048). Both intraoperative cerebral swelling (p = 0.0017) and postoperative GCS score (p < 0.0001) were significantly associated with a raised ICP. Variables such as patient age, aneurysm size, symptomatic vasospasm, intraoperative aneurysm rupture, and secondary cerebral insults such as hypoxia were not associated with raised ICP. Increased ICP adversely affected outcome: 71.9% of patients with normal ICP demonstrated favorable 6-month outcomes postoperatively, whereas 63.5% of patients with ICP between 20 and 50 mm Hg and 33.3% with ICP greater than 50 mm Hg demonstrated favorable outcomes. Among 21 patients whose raised ICP did not respond to mannitol therapy, all experienced a poor outcome and 95.2% died. Among 145 patients whose elevated ICP responded to mannitol, 66.9% had a favorable outcome and only 20.7% were dead 6 months after surgery (p < 0.0001). According to results of multivariate analysis, however, ICP was not an independent outcome predictor (odds ratio 1.26, 95% confidence interval 0.28-5.68). CONCLUSIONS: Increased ICP is common after SAH, even in patients with a good clinical grade. Elevated ICP post-SAH is associated with a worse patient outcome, particularly if ICP does not respond to treatment. This association, however, may depend more on the overall severity of the SAH than on ICP alone. PMID- 15352598 TI - Impact of stereotactic hematoma evacuation on activities of daily living during the chronic period following spontaneous putaminal hemorrhage: a randomized study. AB - OBJECT: Stereotactic evacuation of hematoma has been reported to reduce the incidence of mortality and to improve functional outcome in patients with spontaneous putaminal hemorrhage. Stereotactic evacuation of hematoma has not been widely accepted as a standard therapy, however, because its effect on functional outcome has been regarded as marginal and there have been no randomized trials with sufficient statistical power to quantify the benefits of this procedure. The authors reassessed the value of stereotactic evacuation of hematoma by analyzing its impact on activities of living during the chronic period following spontaneous putaminal hemorrhage in a randomized study. METHODS: Four hundred ninety patients were entered into the study. The severity of their hemorrhages was graded neurologically on admission (neurological grades: 1, eyes are open; 2, eyes are closed but open to weak stimuli; 3, eyes are closed but open to strong stimuli; 4, eyes do not open but extremities move to stimuli; and 5, eyes do not open and extremities do not move to stimuli). Patients with Grade 2 and those with Grade 3 were randomized into two groups with different treatment protocols (Group I, stereotactic evacuation of the hematoma; and Group II, conservative treatment). Patients assigned neurological Grade 4 or 5 were excluded from the study because a large-scale retrospective study in Japan revealed that surgical treatment in patients assigned to these neurological grades does not improve functional outcome. Among the 490 patients, 242 were randomized strictly. This patient population comprised 148 men and 94 women ranging in age from 38 to 80 years (mean 60.5 years). Compared with Group II, Group I treatment resulted in a lower mortality rate and better recovery to functional independence in patients with neurological Grade 3. In patients with Grade 2, Group I treatment contributed to a better recovery of functional outcome and a lower mortality rate, but the difference was not significant. Multivariate analysis confirmed that stereotactic evacuation of the hematoma was contributory to a better recovery in functional outcome. CONCLUSIONS: Stereotactic evacuation of hematoma is clearly of value in selected patients with spontaneous putuminal hemorrhage, whose eyes are closed but will open in response to strong stimuli (neurological Grade 3) on admission. PMID- 15352599 TI - Quantitative analysis of radioisotope cisternography in the diagnosis of intracranial hypotension. AB - OBJECT: The authors attempted a quantitative analysis of conventional radioisotope cisternography for the purpose of more accurate diagnosis of intracranial hypotension. METHODS: Fifty-seven patients suspected of having intracranial hypotension underwent radioisotope cisternography. Whole-body images were obtained 2.5, 6, and 24 hours after intrathecal injection of 111In diethylenetriamine pentaacetic acid. Radioactivity in the cerebrospinal fluid (CSF) space was counted during scanning, and radioisotope clearance was studied. Direct signs of radioisotope leakage into the spinal epidural space were found in 25 patients. Most leaks were located in the lumbosacral region. Analysis of the radioisotope clearance curve revealed two different patterns. In patients without a radiographically demonstrated radioisotope leak, absolutely exponential curves were observed (R2 > 0.99). The activity of the radioisotope decreased at a rate of e(-003) to e(-0.107) (mean +/- standard deviation, e(-0.056 +/- 0.018); 32 patients). Clearance in patients with an overt radioisotope leak was not a simple exponential curve; it could be divided into an early rapid phase and a late slow phase. The clearance rate during the first 6 hours was e(-0.219 +/- 0.127) (25 patients) and e(-0.076 +/- 0.021) thereafter. The authors speculated that the early escape of undiluted radioisotope solution through an aberrant CSF outlet, such as a traumatic spinal dural tear, was responsible for this phenomenon. CONCLUSIONS: The quantitative analysis featured in this study seems to be useful in the diagnosis of intracranial hypotension. A small CSF leak below the limit of radioisotope cisternography resolution might be detected using this technique. PMID- 15352600 TI - Detailed anatomy of the motor and sensory roots of the trigeminal nerve and their neurovascular relationships: a magnetic resonance imaging study. AB - OBJECT: The trigeminal nerve conducts both sensory and motor impulses. Separate superior and inferior motor roots typically emerge from the pons just anterosuperomedial to the entry point of the sensory root, but to date these two motor roots have not been adequately displayed on magnetic resonance (MR) images. The specific aims of this study, therefore, were to identify the superior and inferior motor roots, to describe their exact relationship to the sensory root, and to assess the neurovascular relationships among all three roots of the trigeminal nerve. METHODS: Thirty-three patients and seven cadaveric specimens (80 sides) were studied using three-dimensional (3D) Fourier transform constructive interference in steady-state (CISS) imaging. The 33 patients were also studied by obtaining complementary time-of-flight (TOF) MR angiography sequences with and without contrast enhancement. At least one motor root was identified in all sides examined: in 51.2% of the sides a single motor root, in 37.5% two motor roots, and in 11.2% three motor roots. The superior cerebellar artery (SCA) and the anterior inferior cerebellar artery (AICA) contacted the sensory root in 45.5% of patients and 42.9% of specimens. The SCA often contacted the superior motor root (48.5% of patients and 50% of specimens) and less frequently the inferior motor root (26.5% of patients and 20% of specimens). CONCLUSIONS: Three-dimensional CISS and complementary 3D TOF MR angiography sequences reliably display sensory, superior motor, and inferior motor roots of the trigeminal nerve and their relationships to the SCA and AICA. PMID- 15352601 TI - Normobaric hyperoxia--induced improvement in cerebral metabolism and reduction in intracranial pressure in patients with severe head injury: a prospective historical cohort-matched study. AB - OBJECT: The effect of normobaric hyperoxia (fraction of inspired O2 [FIO2] concentration 100%) in the treatment of patients with traumatic brain injury (TBI) remains controversial. The aim of this study was to investigate the effects of normobaric hyperoxia on five cerebral metabolic indices, which have putative prognostic significance following TBI in humans. METHODS: At two independent neurointensive care units, the authors performed a prospective study of 52 patients with severe TBI who were treated for 24 hours with 100% FIO2, starting within 6 hours of admission. Data for these patients were compared with data for a cohort of 112 patients who were treated in the past; patients in the historical control group matched the patients in our study according to their Glasgow Coma Scale scores after resuscitation and their intracranial pressure within the first 8 hours after admission. Patients were monitored with the aid of intracerebral microdialysis and tissue O2 probes. Normobaric hyperoxia treatment resulted in a significant improvement in biochemical markers in the brain compared with the baseline measures for patients treated in our study (patients acting as their own controls) and also compared with findings from the historical control group. In the dialysate the glucose levels increased (369.02 +/- 20.1 micromol/L in the control group and 466.9 +/- 20.39 micromol/L in the 100% O2 group, p = 0.001), whereas the glutamate and lactate levels significantly decreased (p < 0.005). There were also reductions in the lactate/glucose and lactate/pyruvate ratios. Intracranial pressure in the treatment group was reduced significantly both during and after hyperoxia treatment compared with the control groups (15.03 +/- 0.8 mm Hg in the control group and 12.13 +/- 0.75 mm Hg in the 100% O2 group, p < 0.005) with no changes in cerebral perfusion pressure. Outcomes of the patients in the treatment group improved. CONCLUSIONS: The results of the study support the hypothesis that normobaric hyperoxia in patients with severe TBI improves the indices of brain oxidative metabolism. Based on these data further mechanistic studies and a prospective randomized controlled trial are warranted. PMID- 15352602 TI - Risk of persistent cranial nerve injury after carotid endarterectomy. AB - OBJECT: Cranial nerve injuries, particularly motor nerve injuries, following carotid endarterectomy (CEA) can be disabling and therefore patients should be given reliable information about the risks of sustaining such injuries. The reported frequency of cranial nerve injury in the published literature ranges from 3 to 23%, and there have been few series in which patients were routinely examined before and after surgery by a neurologist. METHODS: The authors investigated the risk of cranial nerve injuries in patients who underwent CEA in the European Carotid Surgery Trial (ECST), the largest series of patients undergoing CEA in which neurological assessment was performed before and after surgery. Cranial nerve injury was assessed and recorded in every patient and persisting deficits were identified on follow-up examination at 4 months and 1 year after randomization. Risk factors for cranial nerve injury were examined by performing univariate and multivariate analyses. There were 88 motor cranial nerve injuries among the 1739 patients undergoing CEA (5.1% of patients; 95% confidence interval [CI] 4.1-6.2). In 23 patients, the deficit had resolved by hospital discharge, leaving 3.7% of patients (95% CI 2.9-4.7) with a residual cranial nerve injury: 27 hypoglossal, 17 marginal mandibular, 17 recurrent laryngeal, one accessory nerve, and three Homer syndrome. In only nine patients (0.5%; 95% CI 0.24-0.98) the deficit was still present at the 4-month follow-up examination; however, none of the persisting deficits resolved during the subsequent follow up. Only duration of operation longer than 2 hours was independently associated with an increased risk of cranial nerve injury (hazard ratio 1.56, p < 0.0001). CONCLUSIONS: The risk of motor cranial nerve injury persisting beyond hospital discharge after CEA is approximately 4%. The vast majority of neurological deficits resolve over the next few months, however, and permanent deficits are rare. Nevertheless, the risk of cranial nerve injury should be communicated to patients before they undergo surgery. PMID- 15352603 TI - Functional separation of languages in the bilingual brain: a comparison of electrical stimulation language mapping in 25 bilingual patients and 117 monolingual control patients. AB - OBJECT: The aim of this investigation was to address three questions in bilingualism research: 1) are multiple languages functionally separated within the bilingual brain; 2) are these languages similarly organized; and 3) does language organization in bilinguals mirror that in monolinguals? 9: During awake dominant-hemisphere craniotomy in each of 25 bilingual patients, the authors mapped both languages by using identical object-naming stimuli. Essential sites for primary (L1) and secondary (L2) languages were compared. Sites were photographically recorded and plotted onto an anatomically referenced grid system. Language organization in bilinguals was then compared with that in 117 monolinguals and 11 monolingual children. CONCLUSIONS: The authors found distinct language-specific sites as well as shared sites that support both languages. The L1 and L2 representations were similar in total cortical extent but significantly different in anatomical distribution. The L2-specific sites were located exclusively in the posterior temporal and parietal regions, whereas the L1 and shared sites could be found throughout the mapped regions. Bilinguals possessed seven perisylvian language zones, in which L2 sites were significantly underrepresented when compared with the distribution of language sites in monolinguals. These L2-restricted zones overlapped the primary language areas found in monolingual children, indicating that these zones become dedicated to L1 processing. These findings support three conclusions. First, it is necessary to map both languages in bilinguals because L1 and L2 sites are functionally distinct. Second, differences exist in the organization of L1 and L2 sites, with L2-specific sites located exclusively in the posterior temporal and parietal lobes. Third, language organization comparisons in bilingual and monolingual brains demonstrate the presence of L2-restricted zones, which are dedicated to L1. PMID- 15352604 TI - Variability of intraoperative electrocortical stimulation mapping parameters across and within individuals. AB - OBJECT: Electrocortical stimulation mapping is regarded as the gold standard of intraoperative mapping for predicting functional outcomes. Nevertheless, methodologies across institutions are inconsistent. Although many vary and maximize stimulation currents at each cortical site, some use a single current level to map the entire exposed cortex. The former comes at the cost of possibly inducing additional afterdischarge activity. The authors retrospectively reviewed their eperience with intraoperative electrocortical stimulation mapping to characterize variability of both mapping and afterdischarge thresholds. METHODS: Seventeen patients satisfied the study inclusion criteria. Significant variability in mapping thresholds was identified within individuals and across the patient population. Moreover, a statistically significant difference in mapping thresholds was demonstrated between the frontal and parietal/temporal lobes (p = 0.007, one-way analysis of variance). The authors report a surprisingly high incidence of afterdischarge during mapping, wide variability in afterdischarge thresholds within individuals and across the study population, and mapping thresholds regularly exceeding afterdischarge thresholds in neighboring cortex. Differences in afterdischarge thresholds across lobes only approached significance (p = 0.086). CONCLUSIONS: To maximize identification of eloquent cortices in some clinical situations, it may be advantageous to maximize currents at each cortical site regardless of adjacent afterdischarge threshold rather than to map the entire exposed cortex at a single current level. Moreover, the current findings highlight the need for electrocorticography during electrocortical stimulation mapping, both to identify when afterdischarges occur and to verify stimulation by recording stimu lation artifacts. The advantages and limitations of maximizing currents at each cortical site as well as mapping at a single current level are discussed. PMID- 15352605 TI - Proton magnetic resonance spectroscopy imaging in the evaluation of patients undergoing gamma knife surgery for Grade IV glioma. AB - OBJECT: The purpose of this study was to assess the differences in spatial extent and metabolic activity in a comparison of a radiosurgical target defined by conventional strategies that utilize the enhancing lesion and a metabolic lesion defined by proton magnetic resonance spectroscopy (MRS) imaging. The authors evaluated whether these differences manifest themselves in the clinical outcome of patients and assessed the value of incorporating 1H-MRS imaging-derived spatial information into the treatment planning process for gamma knife surgery (GKS). METHODS: Twenty-six patients harboring Grade IV gliomas who had previously been treated with external-beam radiation therapy were evaluated by comparing the radiosurgically treated lesion volume with the volume of metabolically active tumor defined on 1H-MRS imaging. The cohort was evenly divided into two groups based on the percentage of overlap between the radiosurgical target and the metabolic lesion volumes. Patients with a percentage of overlap greater than 50% with respect to the metabolic lesion volume were classified as low risk and those with an overlap less than 50% were classified as high risk. Kaplan-Meier estimators were calculated using time to progression and survival as dependent variables. The metabolite levels within the metabolic lesion were significantly greater than those within the radiosurgical target (p < or = 0.001). The median survival was 15.7 months for patients in the low-risk group and 10.4 months for those in the high-risk group. This difference was statistically significant (p < 0.01). CONCLUSIONS: Analysis of the results of this study indicates that patients undergoing GKS may benefit from the inclusion of 1H-MRS imaging in the treatment planning process. PMID- 15352606 TI - Combined use of 18F-fluorodeoxyglucose and 11C-methionine in 45 positron emission tomography-guided stereotactic brain biopsies. AB - OBJECT: The aim of this study was to compare the contribution of the tracers 11C methionine (Met) and 18F-fluorodeoxy-glucose (FDG) in positron emission tomography (PET)-guided stereotactic brain biopsy. METHODS: Forty-five patients underwent combined Met-PET and FDG-PET studies associated with computerized tomography (CT)- or magnetic resonance (MR)-guided stereotactic biopsy. Each patient presented with a lesion that was in proximity to the cortical or subcortical gray matter. The Met-PET and FDG-PET scans were analyzed to determine which tracer offers the best information to guide at least one stereotactic biopsy trajectory. Histologically based diagnoses were rendered in all patients (39 tumors, six nontumorous lesions) and biopsies were performed in all tumors with the aid of PET guidance. When tumor FDG uptake was higher than that in the gray matter (18 tumors), FDG was used for target definition. When FDG uptake was absent or equivalent to that in the gray matter (21 tumors), Met was used for target definition. Parallel review of all histological and imaging data showed that all tumors had an area of abnormal Met uptake and 33 had abnormal FDG uptake. All six nontumorous lesions had no Met uptake and biopsies were performed using CT or MR guidance only. All tumor trajectories had an area of abnormal Met uptake; all nondiagnostic trajectories in tumors had no abnormal Met uptake. CONCLUSIONS: When FDG shows limitations in target selection, Met is a good alternative because of its high specificity in tumors. Moreover, in the context of a single-tracer procedure and regardless of FDG uptake, Met is a better choice for PET guidance in neurosurgical procedures. PMID- 15352607 TI - Comparison of the transvermian and telovelar approaches to the fourth ventricle. AB - OBJECT: The two most common surgical routes to the fourth ventricle are the transvermian and telovelar approaches. The purpose of this study was to compare the microanatomy and exposures gained through these approaches. METHODS: Ten formalin-fixed specimens were dissected in a stepwise manner to simulate the transvermian and telovelar surgical approaches. Stealth image guidance was used to compare the exposures and working angles obtained using these approaches. The transvermian and telovelar approaches provided access to the entire rostrocaudal length of the fourth ventricle floor from the aqueduct to the obex. In addition, both approaches provided access to the entire width of the floor of the fourth ventricle. The major difference between the two approaches regarded the exposure of the lateral recess and the foramen of Luschka. The telovelar, but not the transvermian, approach exposed the lateral and superolateral recesses and the foramen of Luschka. The transvermian approach, which offered an incision through at least the lower third of the vermis, afforded a modest increase in the operator's working angle compared with the telovelar approach when accessing the rostral half of the fourth ventricle. CONCLUSIONS: The transvermian approach provides slightly better visualization of the medial part of the superior half of the roof of the fourth ventricle. The telovelar approach, which lacks incision of any part of the cerebellum, provides an additional exposure to the lateral recesses and the foramen of Luschka. PMID- 15352608 TI - Effects of hyperbaric oxygen therapy on cerebral oxygenation and mitochondrial function following moderate lateral fluid-percussion injury in rats. AB - OBJECT: In the current study, the authors examined the effects of hyperbaric O2 (HBO) following fluid-percussion brain injury and its implications on brain tissue oxygenation (PO2) and O2 consumption (VO2) and mitochondrial function (redox potential). METHODS: Cerebral tissue PO2 was measured following induction of a lateral fluid-percussion brain injury in rats. Hyperbaric O2 treatment (100% O2 at 1.5 ata) significantly increased brain tissue PO2 in both injured and sham injured animals. For VO2 and redox potential experiments, animals were treated using 30% O2 or HBO therapy for 1 or 4 hours (that is, 4 hours 30% O2 or 1 hour HBO and 3 hours 100% O2). Microrespirometer measurements of VO2 demonstrated significant increases following HBO treatment in both injured and sham-injured animals when compared with animals that underwent 30% O2 treatment. Mitochondrial redox potential, as measured by Alamar blue fluorescence, demonstrated injury induced reductions at 1 hour postinjury. These reductions were partially reversed at 4 hours postinjury in animals treated with 30% O2 and completely reversed at 4 hours postinjury in animals on HBO therapy when compared with animals treated for only 1 hour. CONCLUSIONS: Analysis of data in the current study demonstrates that HBO significantly increases brain tissue PO2 after injury. Nonetheless, treatment with HBO was insufficient to overcome injury-induced reductions in mitochondrial redox potential at 1 hour postinjury but was able to restore redox potential by 4 hours postinjury. Furthermore, HBO induced an increase in VO2 in both injured and sham-injured animals. Taken together, these data demonstrate that mitochondrial function is depressed by injury and that the recovery of aerobic metabolic function may be enhanced by treatment with HBO. PMID- 15352609 TI - Application of p27 gene therapy for human malignant glioma potentiated by using mutant p27. AB - OBJECT: Malignant glioma could be an ideal candidate for local gene therapy because its invasion is local and it has little metastatic potential. A low expression level and high degradation activity of p27 are known to constitute an independent poor prognostic factor in patients with malignant glioma. In this study, the authors investigated the roles of wild-type p27 and mutant p27 on the treatment of malignant glioma. METHODS: The authors used two adenoviruses: one expressed wild-type p27 (ad-p27wt) and the other, containing a mutation at the major metabolic site, expressed mutant p27 (ad-p27mt). The antitumor effects of the two adenoviruses were compared with respect to cell growth arrest, cell cycle alteration, apoptosis induction, and in vitro tumorigenicity in three glioblastoma mutiforme (GBM) cell lines and in a primary GBM cell line. Transduction with ad-p27wt or ad-p27mt induced the production of p27 and the dephosphorylation of pRB. The protein level of mutant p27 was significantly higher than that of wild-type p27. The ad-p27wt induced cell cycle arrest at the G1-S transition point, whereas the ad-p27mt induced arrest at the G2-M point. Both ad-p27wt and ad-p27mt induced a growth-inhibiting effect, apoptosis, and suppression of in vitro tumorigenicity; however, ad-p27mt displayed a stronger antitumor effect than ad-p27wt in brain tumor cell lines. CONCLUSIONS: Gene therapy involving p27, especially mutant p27, has the potential to become a novel and powerful therapy for malignant glioma. PMID- 15352610 TI - Neurotransmitter release from high-frequency stimulation of the subthalamic nucleus. AB - OBJECT: High-frequency stimulation (HFS) delivered through implanted electrodes in the subthalamic nucleus (STN) has become an established treatment for Parkinson disease (PD). The precise mechanism of action of deep brain stimulation (DBS) in the STN is unknown, however. In the present study, the authors tested the hypothesis that HFS within the STN changes neuronal action potential firing rates during the stimulation period by modifying neurotransmitter release. METHODS: Intracellular electrophysiological recordings were obtained using sharp electrodes in rat STN neurons in an in vitro slice preparation. A concentric bipolar stimulating electrode was placed in the STN slice, and electrical stimulation (pulse width 50-100 microsec, duration 100-2000 msec, amplitude 10 500 microA, and frequency 10-200 Hz) was delivered while simultaneously obtaining intracellular recordings from an STN neuron. High-frequency stimulation of the STN either generated excitatory postsynaptic potentials (EPSPs) and increased the action potential frequency or it generated inhibitory postsynaptic potentials and decreased the action potential frequency of neurons within the STN. These effects were blocked after antagonists to glutamate and gamma-aminobutyric acid were applied to the tissue slice, indicating that HFS resulted in the release of neurotransmitters. Intracellular recordings from substantia nigra pars compacta (SNc) dopaminergic neurons during HFS of the STN revealed increased generation of EPSPs and increased frequency of action potentials in SNc neurons. CONCLUSIONS: During HFS of STN neurons the mechanism of DBS may involve the release of neurotransmitters rather than the primary electrogenic inhibition of neurons. PMID- 15352611 TI - Suprapineal recess: an alternate site for third ventriculostomy? Case report. AB - This 30-year-old woman presented with clinical symptoms and signs of intracranial hypertension and Parinaud syndrome secondary to ventriculoperitoneal shunt dysfunction. Magnetic resonance (MR) imaging revealed gross triventricular hydrocephalus with a large suprapineal recess due to aqueductal stenosis. Using an endoscopic approach, a ventriculostomy was performed within the floor of the dilated suprapineal recess. Following this procedure the patient experienced alleviation of all her neurological symptoms and signs. Postoperative MR imaging and cerebrospinal fluid flow studies demonstrated a functioning ventriculostomy. The anatomy of the suprapineal recess and its suitability for endoscopic ventriculostomy are discussed. PMID- 15352612 TI - Diagnosis of Erdheim-Chester disease by using computerized tomography-guided stereotactic biopsy of a caudate lesion. Case report. AB - The authors present the case of a 27-year-old woman with Erdheim-Chester disease (ECD) and extensive intracranial involvement, in whom the initial diagnosis of ECD was established based on computerized tomography (CT)-guided stereotactic biopsy of a caudate lesion. Erdheim-Chester disease is a rare non-Langerhans cell histiocytosis of unknown origin that is clinically characterized by bone pain, diabetes insipidus, and exophthalmos. The radiological hallmarks of the disease include symmetrical sclerosis of the long bones with epiphysial sparing and increased tracer uptake in lesions seen on scintigraphic imaging. Erdheim-Chester disease is characterized histologically by the presence of infiltrating lipid laden histiocytes that commonly involve the retroperitoneum, orbits, skin, pericardium, lungs, and long bones. Although the occurrence of diabetes insipidus often precedes the diagnosis of ECD by more than a decade in most patients, magnetic resonance imaging- and CT-documented central nervous system involvement is exceedingly rare. In the setting of neurological involvement, neurosurgical biopsy has been reported seven times in the literature, with only one of these biopsies being the basis for the initial diagnosis of the disease. The authors' case represents only the second time the disease has been diagnosed by means of neurosurgical biopsy, highlighting the diagnostic difficulties that patients with EDC present. Skeletal radiographs were confirmatory in this case and this modality should be emphasized as the simplest and most direct route to the diagnosis. The degree of neurological involvement further distinguishes the case presented from prior reports in the literature. The multiple bilateral intraaxial lesions were intensely enhancing on contrast CT scans, distributed infra- and supratentorially, involving both white and gray matter, and associated with diffuse cerebral edema. The case presented is also remarkable by virtue of the symmetrical involvement of the caudate nuclei, representing the first such example documented in the literature. The diagnosis, treatment, and outcome in this patient are discussed, and a review of the literature is presented. PMID- 15352613 TI - Transition from meningeal melanocytoma to primary cerebral melanoma. Case report. AB - The authors describe the first case of an intracranial transition of a melanocytoma into a primary malignant melanoma within a short time. A 37-year-old woman presented with progressive brainstem syndrome due to a tumor, originally diagnosed and treated 12 years earlier, that extended from the petroclival area to the anterior craniocervical junction. The histological workup following subtotal tumor resection of the initial tumor had revealed the typical features of a fibrous melanocytic meningioma without increased proliferation. Ten years after the patient had completed treatment for the melanocytic meningioma, control neuroimaging demonstrated growth of the residual tumor with compression of the brainstem. Another neurosurgical intervention revealed a dark tumor of hard consistency. At this time immunohistochemical examinations demonstrated melanocytic features (expression of vimentin, S100 protein, and melan A) of the lesion with focally increased proliferation (5% of Ki-67-positive cells) but no higher mitotic activity. Clinical signs of deterioration along with imaging confirmed tumor progression precipitated another operation within 7 months. A neuropathological examination revealed epithelial and anaplastic changes and indicated that the MIB-1 indices were greater than 25%. Pleomorphic changes and a focal high mitotic activity led to the diagnosis of a primary cerebral malignant melanoma. The patient's later clinical course consisted of a rapid diffuse meningeal spread of the lesion throughout the entire brain and spine. Despite whole-brain and stereotactic radiation therapy as well as chemotherapy, the patient died 4 months after the last neuropathological diagnosis. Although grossly resembling a meningioma, melanocytomas lack the former's histological and immunohistochemical features. The biological behavior of a melanocytoma is variable and recurrence may happen after subtotal resection, but intracranial transition into a malignant melanoma has not been observed previously. PMID- 15352614 TI - Ruptured aneurysm arising from a basilar artery fenestration and associated with a persistent primitive hypoglossal artery. Case report and review of the literature. AB - A 42-year-old woman experienced the sudden onset of a severe headache. Angiograms demonstrated a persistent primitive hypoglossal artery (PHA) originating from the internal carotid artery at the C-2 vertebral level. In addition, a fenestration at the PHA-basilar artery (BA) junction and an aneurysm at the proximal end of this fenestration were revealed. To perform endovascular embolization of the aneurysm, a microcatheter was introduced into the aneurysm sac via the PHA and two Guglielmi Detachable Coils were placed in the aneurysm. The patient's postoperative course was uneventful, and she was able to resume her normal life. Although many clinical cases have been reported in which a ruptured aneurysm was associated with a PHA or a BA fenestration, as far as the authors know there has been no case in the literature in which a ruptured aneurysm associated with both anomalies and no case in which endovascular embolization was used to treat a ruptured aneurysm associated with a PHA. This rare case is discussed and a review of the relevant literature is presented. PMID- 15352615 TI - Endotracheal tube electrodes to map and monitor activities of the vagus nerve intraoperatively. Technical note. AB - Difficulty swallowing due to damage of the vagus nerve is one of the most devastating complications of surgery in and around the medulla oblongata; therefore, intraoperative anatomical and functional evaluation of this nerve is crucial. The authors applied endotracheal tube surface electrodes to record electromyography (EMG) activity from vocal cords innervated by the vagus nerve. The vagal nucleus or rootlet was electrically stimulated during surgery and vocalis muscle EMG activities were displayed by auditory and visual signals. This technique was used successfully to identify the vagus motor nerve and evaluate its integrity during surgery. The advantages of this method compared with the use of needle electrodes include safe simple electrode placement and stable recording during surgery. In cases involving a pontine cavernoma pressing the nucleus or a jugular foramen tumor encircling the rootlet, this method would be particularly valuable. Additional studies with a larger number of patients are needed to estimate the significance of this method as a means of functional monitoring to predict clinical function. PMID- 15352616 TI - A quiet hand for microneurosurgery: twiddle your thumb. Technical note. AB - Fatigue is both the most tremor-producing factor and the constant companion of a busy neurosurgeon. Because of the difficulty in controlling tremor-generating factors, the authors sought to develop a means of manipulating surgical instruments with minimal muscle movement while allowing accurate and repeatable movements in all conditions. The "quiet hand technique" is an isolation technique that creates a stable platform by forming the four fingers into a salute that thenceforth moves as one. Fine movements are supplied by the thumb. For instruments that open and close, the thumb abducts and adducts. For suture placement using a round needle holder, the thumb flexes and extends. Because only the thumb moves, the extent of movement is decreased and therefore momentum and inertia are limited as well. Previously, microsurgeons favored ballistic movements. With the quiet hand technique, however, tension movements are easy. The greater control makes tying 10-0 and 11-0 sutures more predictable. Although the quiet hand technique was developed for use in small-vessel anastomosis, the neurophysiological principles on which it is based apply to movement at all levels and are applicable to working a bipolar coagulator, pickups, a vascular clip holder, or other soft-tissue manipulators. PMID- 15352617 TI - Neuritis ossificans of the sciatic nerve. Case illustration. PMID- 15352618 TI - Ictal bradycardia after endovascular treatment of an aneurysm. Case illustration. PMID- 15352619 TI - Harvey W. Cushing and cerebrovascular surgery: Part I, Aneurysms. AB - The development of surgical techniques for the treatment of intracranial aneurysms has paralleled the evolution of the specialty of neurological surgery. During the Cushing era, intracranial aneurysms were considered inoperable and only ligation of the carotid artery was performed. Cushing understood the limitations of this approach and advised the need for a more thorough understanding of aneurysm pathology before further consideration could be given to the surgical treatment of cerebral aneurysms. Despite his focus on brain tumors, Cushing's contributions to the discipline of neurovascular surgery are of great importance. With the assistance of Sir Charles Symonds, Cushing described the syndrome of subarachnoid hemorrhage. He considered inserting muscle strips into cerebral aneurysms to promote aneurysm sac thrombosis and designed the "silver clip," which was modified by McKenzie and later used by Dandy to clip the first intracranial aneurysm. Cushing was the first surgeon to wrap aneurysms in muscle fragments to prevent recurrent hemorrhage. He established the foundation on which pioneers such as Norman Dott and Walter Dandy launched the modern era of neurovascular surgery. PMID- 15352620 TI - Harvey W. Cushing and cerebrovascular surgery: Part II, Vascular malformations. AB - The surgical treatment of cerebrovascular malformations intrigued early neurosurgeons. Cushing defined vascular malformations as tumors arising from cerebral blood vessels. He successfully resected the first arteriovenous malformation 3 years after it had been irradiated. In the absence of angiography, the pathoanatomy of these lesions remained elusive and early techniques such as cortical vein ligation proved catastrophic. Cushing demonstrated the favorable results of radiation treatments on vascular malformations and advocated decompressive craniectomy followed by radiotherapy. He ligated cortical feeding vessels and external carotid arteries with an improved understanding of the angioarchitecture of vascular malformations. He stressed the importance of preoperative diagnosis because the radical resection of nonirradiated vascular malformations challenged the limitations of the available neurosurgical armamentarium. PMID- 15352621 TI - Motor evoked potentials. PMID- 15352622 TI - Human serum albumin. PMID- 15352623 TI - High-dose mannitol. PMID- 15352624 TI - High-dose mannitol. PMID- 15352625 TI - High-dose mannitol. PMID- 15352626 TI - [Consequence of intellectual property and consortium project in neurosurgery]. PMID- 15352627 TI - [Differences in outcome in patients with poor grade subarachnoid hemorrhage (WFNS grade IV and V): according to the perioperative management against vasospasm]. AB - Cerebral vasospasm remains a leading cause of morbidity and increasing mortality rates following aneurysmal subarachnoid hemorrhage (SAH). The rate of vasospasm and the outcome (Glasgow Outcome Scale; GOS) especially in poor WFNS grade were retrospectively analyzed over a 6-year period. Patients were divided into three different groups. The first group (pre-group) consisted of 43 patients (grade IV: 31, grade V: 12), who were admitted between 1996 and 1998. When vasospasm occurred, they were mainly treated by papaverine (PPV) and percutaneous transluminal angioplasty (PTA). In the second group (mil-cis group), for the prevention of vasospasm, cisternal irrigation therapy with milrinone was applied in 24 patients (grade IV: 13, grade V: 11), who were admitted in the period between 1999 and 2001. The third group, (w/o mil-cis group), consisted of 30 patients (grade IV: 15, grade V: 15), in whom cisternal irrigation therapy was not able to be carried out. In grade IV, vasospasm was observed in 66% of the patients the first group, 50% in the w/o mil-cis group and significantly less in the mil-cis group (15%, p<0.024). In grade V, the rate of vasospasm was also lower in the mil-cis group but no statistical significance was revealed. Although the rate of favorable outcome in GOS was highest and the rate of death was least in the mil-cis group in both grade IV and V, only the trend was observed. Many factors should be considered, Cisternal irrigation therapy with milrinone reduced the occurrence of vasospasm. However, outcome was not improved because of the initial poor clinical condition. PMID- 15352628 TI - [Unruptured cerebral aneurysms; the features of cases undetected with MR angiography]. AB - With the development of MRI and MRA, many unruptured aneurysms have been detected and treated. Nevertheless, not a few false-positive and false-negative cases are found. We investigate aneurysms that were suspected after screening MRA at the neurosurgical outpatient clinic and the features of aneurysms detected not with MRA but with DSA were studied. Seventy-six patients (85 aneurysms) were suspected due to screening MRA and DSA was performed in 64 (71 aneurysms) of them. Correct diagnosis of cerebral aneurysms with MRA was obtained in 44 patients (45 aneurysms, 63.4%), while false-negative cases were found in 17 patients (plus 20 aneurysms) and false-positive cases in 7 patients (10 aneurysms). The accuracy was 97.2% in ACA, 93% in MCA, 94.4% in VA-BA, and 78.9% in IC, while the sensitivity 100%, 88.2%, 81.8%, 64.7% and the specificity 96.5%, 94.4%, 96.7%, 91.9%, respectively. The features of aneurysms correctly diagnosed with MRA were relatively large ACA, including AcoA, MCA and VA-BA aneurysms, whereas the features of aneurysms undetected with MRA were small IC aneurysms (1-3mm in diameter), especially at the C2-3 portion. These aneurysms at the C2-3 portion or at unusual portions tended to be difficult to detect even with 3D-CTA. Though most of the aneurysms detected with DSA but not with MRA tended to be small and not interventionally treated in the present study, we should pay attention to the fact that these aneurysms are overlooked despite the possibility that they may become enlarged or rupture. Though ruptured aneurysms were surgically treated with only MRA or 3D-CTA without conventional angiography in these days, we recommend the examination of the unruptured cases, which are usually asymptomatic and not hasty, with precise inspection by target MIP, high-performance 3D-CTA or DSA. PMID- 15352629 TI - [Successful trans-arterial occlusion of a vertebral arteriovenous fistula with aid of intra vascular ultra sound (IVUS) and balloon catheter]. AB - We report a case of spontaneous vertebral arteriovenous fistula manifesting pulsating tinnitus with left cervical bruit. A 50-year-old woman presented with pulsatile tinnitus of unknown duration. Angiography revealed a simple and direct fistula between the second segment of the left vertebral artery and the epidural venous system. The fistula was well visualized by intra vascular ultra sound (IVUS). Balloon catheters were used for regulating blood flow during coil embolization, which successfully obliterated the fistula, with subsequent control of the symptoms. Postoperative course (18 months) was uneventful. PMID- 15352630 TI - [Two cases of spinal arteriovenous malformation presenting with subarachnoid hemorrhage]. AB - Two cases of spinal arteriovenous malformation (AVM) with subarachnoid hemorrhage (SAH) are reported. The first case is that of a 14-year-old boy who was transferred to our hospital with a sudden onset of headache. Neurological examination revealed no motosensory deficit, but a brain CT showed a slight diffuse SAH. A left vertebral angiogram demonstrated intramedullary AVM in the cervical region of the spinal cord. This AVM was therefore occluded using a solid embolization material. The patient was then discharged without neurological deficit. The second case is that of a 67-year-old man who visited our hospital with a sudden onset of headache. Neurological examination revealed no motor or sensory deficit, but a brain CT showed SAH, which was dominant in the posterior fossa. Initial cerebral angiography demonstrated no abnormality such as cerebral aneurysm or AVM except for laterality of the C1 radiculo-meningeal artery. A second angiogram on day 11 demonstrated spinal arteriovenous fistula (AVF), which was fed by the left radiculo-meningeal artery and drained to the posterior spinal vein. Embolization for the AVF was performed using liquid material. He was then discharged without neurological deficit. These two cases revealed non-specific SAH symptoms and were indistinguishable from other ruptured aneurysms. Although the brain CT can show a slight SAH or posterior fossa dominant SAH, repeated angiography may be necessary to verify and conclude the diagnosis of spinal AVM. PMID- 15352631 TI - [Therapeutic effect of spinal cord stimulation for a patient suffering spasticity after hypoxia of the brain]. AB - We reported a case of severe spasticity of the bilateral upper and lower limbs which was improved by cervical spinal cord stimulation (SCS). A 53-year-old man was suffering from sudden cardiopulmonary arrest while walking. After receiving cardiopulmonary resuscitation for 40 minutes, his cardiopulmonary function re started, but he failed to regain consciousness. MRI revealed a hypoxic brain in his bilateral basal ganglia and occipital lobes. After 2 months, his consciousness advanced to a vegetative state and the muscle tone of his bilateral upper and lower limbs deteriorated to severe spasticity. SCS was performed in the expectation of improving his consciousness. Contrary to this anticipated result, his vegetative state continued but the severe spasticity of his upper and lower limbs improved during the SCS. Single photon emission computed tomography (SPECT) during the period of stimulation revealed a high blood flow, especially in the area of the basal ganglia, thalamus, brain stem and cerebellum, compared with off stimulation blood flow. The neurophysiologic mechanisms of these abnormal fields and the underlying aberrant afferent nerve impulses from the posterior funiculus in the cervical cord to the cerebral sensory cortex, which may indeed be secondary to ischemic brain, may be regulated by SCS, also adding the effect of increased blood flow to the brain. PMID- 15352632 TI - [Supratentorial primitive neuroectodermal tumor: report of a surgical case]. AB - We reported a rare case of supratentorial primitive neuroectodermal tumors (sPNET) and reviewed the literature. A 15-year-old girl presented with a one month history of headache and vomiting. CT scans showed a huge, solid and cystic tumor with calcification, occupying the left anterior cranial fossa. The solid portion of the tumor was hypointense on T1-weighted images, slightly hyperintense on T2-weighted images, hyperintense on diffusion- weighted images, isointense on fluid-attenuated inversion recovery (FLAIR) images, and strongly enhanced after administration of contrast medium. The expansile tumor had a broad attachment to the dura matter of the anterior cranial fossa. The patient underwent an uneventful extirpation of the tumor. Microscopically, the solid tumor contained small, round poorly-differentiated cells with pleomorphic nuclei and brisk mitotic activity. The tumor cells were immunoreactive for synaptophysin and GFAP, whereas lack of MIC2 gene product expression was confirmed using the monoclonal antibody 12E7. The MIB-1-positive rate was 43%. The histological diagnosis was sPNET. Postoperatively, a dose of 30.6Gy was administrated to the whole brain and a boost of 19.6Gy to the T2-hyperintensity lesion. The patient successively received a dose of 30.4Gy to the whole spine and Linac stereotactic radiotherapy with a marginal dose of 16.8Gy at the tumor bed. Three months after radiotherapy the patient received chemotherapy using carboplatin and etoposide. Follow-up MR images showed no evidence of recurrent tumor 5 months after the radiochemotherapy. On the basis of MR findings on both diffusion-weighted and FLAIR images, preoperative diagnosis of sPNET may be important for choosing appropriate therapeutic strategies for this tumor. PMID- 15352633 TI - [Stent-assisted percutaneous transluminal angioplasty for intracranial vertebral and basilar artery stenosis: two case reports]. AB - We report two cases of patients who underwent stent-assisted percutaneous transluminal angioplasty for intracranial vertebral artery (VA) and basilar artery (BA) stenosis. They presented with sudden onset of vertigo and right hemiparesis. Vertebral angiographies revealed severe proximal BA stenosis (case 1) and intracranial VA stenosis (case 2). They underwent angioplasty and stenting, with excellent angiographic results. The availability of new flexible intravascular stents, allowing access to tortuous proximal intracranial vessels, provides a new therapeutic approach for patients with vertebral and basilar artery stenosis. PMID- 15352634 TI - [Spontaneous regression of primary central nervous system lymphoma: a case report]. AB - A rare case of primary central nervous system lymphoma that regressed spontaneously as shown on serial cranial magnetic resonance imagings (MRI) is described. A 60-year-old woman was admitted to our hospital with a well-enhanced mass lesion in the cerebellum although MRI had demonstrated no abnormal findings 4 months before admission. On admission, She complained of headache, but no neurological deficits were observed. The patient underwent exploratory craniotomy and the tumor was partially removed on February 14. Pathologic examination suspected B-cell type of malignant lymphoma. Postoperative MRI performed on February 23, 2001 showed disappearance of the mass lesion without further treatment. Different polymerase chain reaction (PCR) assays detected clonal immunoglobulin heavy chain gene rearrangements in paraffin-embedded tissues diagnosed as B-cell lymphoma. The patient was discharged without any neurological deficits for two weeks and was followed up on MRI. One year after operation, MRI revealed an abnormal finding in the cerebellum without apparent neurological deterioration. Radiation therapy of the whole brain and the local site was carried out with a total dose of 50Gy. MRI demonstrated complete remission of the tumor following radiation therapy. She was admitted again to our hospital with right hemiparesis on May, 2003. Subsequently, systemic chemotherapy was carried out. The patient died 2 years and 10 months after her initial presentation. PMID- 15352635 TI - [Surgical treatment for cerebral aneurysm after incomplete Guglielmi detachable coil embolization]. AB - The frequency of surgical treatment for intracranial aneurysms that have been incompletely treated by endovascular coiling will increase in time. The authors describe their experience in the following 3 patients. There was one non-ruptured aneurysm and two ruptured ones. The intervals between coiling and surgery were 1, 5, and 10 months. Surgery was indicated because of partial treatment and/or growth of residual neck. Observation during surgery revealed that coils were exposed to the subarachnoid space at the tip of the aneurysm in the cases of ruptured aneurysms. When the residual neck was large enough for the size of the clip blades, there was no need to remove the coils. The coils in the neck were able to be removed when the interval between coiling and surgery was very short, but could not be removed when the interval was prolonged. Intraoperative angiography and reconstructive bypass surgery may be essential in these complex cases. As pointed out previously, the operative approach should be determined by the need for coil removal and the duration since coiling. PMID- 15352636 TI - [Pallidotomy: a guide to stereotactic brain operations]. PMID- 15352637 TI - [Clinico-pathology of skull tumor: Ewing's sarcoma, malignant lymphoma, plasmacytoma/multiple myeloma]. PMID- 15352638 TI - Why do oncologists need this new journal? PMID- 15352639 TI - New therapeutic approaches for hot flashes in women. AB - While most women will suffer from hot flashes at some point over their lifetime, most symptoms resolve with time. However, some women may experience severe and/or long-lasting hot flashes. Estrogen, the most effective treatment for hot flashes, is not generally recommended for women with a history of breast cancer or women at high risk of developing breast cancer. Moreover, long-term administration of estrogen to healthy women is associated with increased risks of cardiovascular disease, stroke, and breast cancer. Newer antidepressants from the selective serotonin or noradrenergic reuptake inhibitor family, such as venlafaxine and paroxetine, appear to be among the most effective nonhormonal agents for the treatment of hot flashes. New information demonstrates that gabapentin also is an effective nonhormonal therapy for hot flashes. In this review, we will discuss current knowledge of the epidemiology and pathophysiology of hot flashes, along with treatment options. We will focus on nonhormonal treatments that have been studied in prospective randomized clinical trials, and will present an algorithm for the treatment of symptomatic patients. PMID- 15352640 TI - Management of dyspnea. AB - Patients with cancer frequently report dyspnea, the uncomfortable awareness of breathing. Lung involvement with cancer does not predict its occurrence. Patients describe it as one of the most frightening and distressing symptoms, and patient self-report is the only reliable measure. Measurements of respiratory rate, oxygen saturation, and arterial blood gases do not measure dyspnea. Opioids in modest doses have been demonstrated to give effective relief of dyspnea, whether or not identifiable reversible causes exist. Medical management of dyspnea can be directed at the underlying cause when the potential benefits outweigh the burdens of such treatment. In rare cases where symptomatic treatment is unable to control dyspnea to the patient's satisfaction, sedation is an effective, ethical option. PMID- 15352641 TI - Understanding and managing chemotherapy-induced diarrhea. AB - Diarrhea is particularly problematic in patients receiving fluoropyrimidines and/or irinotecan. Careful patient monitoring, patient education, and good patient-provider communication are the primary tools of prevention. The patient must be carefully evaluated on a regular basis early in treatment, so that mid course corrections, dose adjustments, or dose delays can be instituted early on if indicated. Diet need not be modified as a preventive measure, but once diarrhea occurs, a number of modifications must be made. Maintenance of fluid intake is critical and inability to maintain adequate hydration would be a primary indication for parenteral fluid support. Oral intake of fluids should not be limited to plain water only, since electrolytes need to be replenished. Early recognition of diarrhea and early pharmacologic intervention can greatly facilitate successful resolution of this treatment complication. PMID- 15352642 TI - Pilot evaluation of citalopram for the relief of hot flashes. AB - Symptoms associated with premature menopause are a significant problem for women with a history of breast cancer who cannot take hormone replacement therapy. Thus, effective nonhormonal alternatives are needed to manage hot flashes, the most prevalent symptom of menopause. Previous studies have defined that venlafaxine, an anti-depressant, is an effective treatment for such hot flashes. Based on suggestive anecdotal information, we set out to evaluate, in a pilot trial, whether the antidepressant citalopram might be a good nonhormonal treatment option to add to our armamentarium for controlling hot flashes. A prospective pilot study was developed in which patients were studied for 5 weeks, with the first week used to establish a baseline, followed by 4 weeks of treatment with citalopram. During the first week of treatment, 10 mg/day of citalopram was taken while 20 mg/day was taken during each of the following three weeks. Hot-flash diaries were completed daily, symptom diaries and quality-of life items were completed weekly and the Profile of Mood States was completed at baseline and at week 5. Evaluable patients who completed the study had a mean hot flash frequency reduction of 58% and a mean hot-flash score reduction of 64% from baseline to week 5. The patients finishing the study also reported decreased anger, tension and depression, as well as improved mood. This pilot trial suggests that citalopram may be an effective non-hormonal treatment for hot flashes in women who can tolerate it. PMID- 15352643 TI - Getting comfortable with methadone. PMID- 15352644 TI - Treatment of opiate-related sedation: utility of the cholinesterase inhibitors. AB - Daytime sedation is a common and potentially dose-limiting side effect of the opiate analgesics. The psychostimulants, such as methylphenidate, are frequently prescribed to treat this problem, but their use may be limited by side effects, such as weight loss, anxiety, or insomnia, or tolerance to their antisedative effects. Medications which enhance intracerebral cholinergic activity may offer an alternative treatment approach, since functional deficits of acetylcholine may, in part, account for the sedative and mind-dulling effects of opiates. Preliminary studies with donepezil, a centrally acting acetylcholinesterase (AChE) inhibitor approved for use in Alzheimer's disease, have suggested at least short-term benefit in treating opiate-related sedation. In a retrospective study, we reviewed the results of donepezil treatment in 40 patients, 37 of whom had cancer, which in most cases was in an advanced stage. All patients were receiving chronic opiate treatment, and the majority were on a stable opiate dose in the 2 weeks preceding the initiation of donepezil. The average opiate dose was 844 mg in oral morphine equivalents. Seventy-three percent of the patients experienced moderate or greater improvement on the Clinical Global Improvement Scale. Evaluations on the Epworth Sleepiness Scale (ESS; 0-24), visual analog sleepiness scale (VAS; 0-100), and average pain levels (0-100), before and after donepezil treatment, were obtained on 19 patients. Prior to donepezil, ESS, VAS, and pain scores were 18.5, 76.3, and 47.4. After an average of 21 days of donepezil treatment, scores were 9.5 (P < 0.001), 39.5 (P < 0.001), and 36.6 (P = 0.07), respectively. The mean total duration of treatment for patients was 54.4 days, with most patients stopping donepezil due to progression to a terminal state. Patients were generally started on 5 mg/ day, but 17 patients required higher doses to achieve or maintain efficacy, the average treatment dose being 9.13 mg/day. We concluded from this study that centrally acting AChE inhibitors are promising agents in the treatment of sedation,and perhaps of other neuropsychological side effects associated with the use of opiate analgesics. PMID- 15352645 TI - Olanzapine and mirtazapine for multiple palliation. PMID- 15352647 TI - Anemia: cancer patients who are very likely to benefit often go untreated. PMID- 15352646 TI - Does palliative chemotherapy palliate? PMID- 15352648 TI - Pulmonary thromboembolism following BMT: what autopsies have shown. PMID- 15352649 TI - Results from the Last Acts campaign: how can we improve? PMID- 15352650 TI - How to use implantable intrathecal drug delivery systems for refractory cancer pain. PMID- 15352651 TI - Imatinib results in better quality of life for CML patients than interferon/Ara C. PMID- 15352652 TI - Pathogenesis-based treatment of chemotherapy-induced nausea and vomiting--two new agents. AB - Chemotherapy-induced nausea and vomiting (CINV) is associated with a significant deterioration in quality of life. The emetogenicity of the chemotherapeutic agents, repeated chemotherapy cycles, and patient risk factors (female gender, younger age, alcohol consumption, history of motion sickness) are the major risk factors for CINV. The use of 5-hydroxytryptamine3 (5-HT3) receptor antagonists plus dexamethasone has significantly improved the control of acute CINV, but delayed nausea and vomiting remains a significant clinical problem. Although the 5-HT3 receptor antagonists, dexamethasone, and metoclopramide have been used to prevent delayed CINV, only dexamethasone appears to have much efficacy with acceptable toxicity. Recent studies have introduced two new agents, palonosetron and aprepitant, for the prevention of both acute and delayed CINV. Palonosetron is a new 5-HT3 receptor antagonist with a longer half life and a higher binding affinity than older 5-HT3 receptor antagonists. It improves the complete response rate (no emesis, no need for rescue) of acute and delayed CINV in patients receiving moderately emetogenic chemotherapy compared to the older 5-HT3 receptor antagonists. The other agent, aprepitant, is the first agent available in the new drug class of neurokinin-1 receptor antagonists. When added to a standard regimen of a 5-HT3 receptor antagonist and dexamethasone in patients receiving highly emetogenic chemotherapy, it improves the complete response rate of acute CINV. Aprepitant also improves the complete response of delayed CINV when compared to placebo and when used in combination with dexamethasone compared to dexamethasone alone. Acute and delayed nausea may also be improved by aprepitant when used in combination with a 5-HT3 and dexamethasone prechemotherapy or with daily dosing for 3-5 days following chemotherapy. Based on these studies, new guidelines for the prevention of CINV are proposed. Future studies may consider the use of palonosetron and aprepitant with current and other new agents (olanzapine, gabapentin) in moderately and highly emetogenic chemotherapy, as well in the clinical settings of multiple-day chemotherapy and bone marrow transplantation. PMID- 15352653 TI - Epoetin alfa and chemotherapy: a summary of two studies on preventive and maintenance use. PMID- 15352654 TI - Mechanisms and management of neuropathic pain in cancer. AB - Neuropathic pain is a common syndrome in people with cancer. The pathophysiology of such pain in cancer is not fully understood, often leading to poor management and needless suffering. Knowledge regarding the potential mechanisms of neuropathic pain, skill in appropriate history-taking and physical-assessment techniques, and awareness of the more common neuropathic pain syndromes and their etiologies, as well as familiarity with the role of new pharmacologic interventions, will allow healthcare professionals to provide better relief of neuropathic pain. At present,a variety of agents are used to treat neuropathic pain situations. Rehabilitation of persons with neuropathic pain should be part of overall management and should address functional impairment and safety factors to prevent accidents resulting from sensory loss. PMID- 15352655 TI - Current status of selenium and other treatments for secondary lymphedema. AB - The unsatisfactory status of lymphedema treatment in patients with cancer warrants an intensified search for new treatment approaches, especially since survival of cancer patients has improved markedly. Lymphedema, regardless of etiology, is essentially incurable, but current therapeutic approaches can reduce swelling, restore shape, and prevent inflammatory episodes. The most conservative approach is physical therapy, including a combination of compression, exercise, and, if possible, massage.Meticulous skin care is needed to avoid recurrent inflammatory episodes. If physical therapy fails, surgery, in the form of reducing or "debulking" operations, is indicated, while microsurgical interventions for certain forms of lymphedema continue to be evaluated. Drug therapy has included the use of diuretics, corticosteroids, and coumarin- or flavonoid-type compounds. Diuretics and corticosteroids may be useful in edema of mixed origin and in palliative circumstances but cannot be recommended for persistent lymphedemas. Coumarin and flavonoids reduce swelling in all types of lymphedema, but their long-term use is problematic. One promising step in drug therapy seems to be the introduction of free-radical scavengers, such as selenium. Present data demonstrate that selenium can enhance the benefits of physical therapy in radiation-induced lymphedemas. The very low toxicity profile of selenium and its cost effectiveness are further arguments for its use in lymphedema treatment. PMID- 15352656 TI - Assessment of hematologic effects and fatigue in cancer patients with chemotherapy-induced anemia given darbepoetin alfa every two weeks. AB - The objective of this ongoing trial is to study the ability of darbepoetin alfa to reverse chemotherapy-induced anemia in cancer patients, and to relate improvement in hemoglobin with changes in fatigue and functional capacity. Eligible subjects had a nonmyeloid malignancy, were receiving multicycle chemotherapy, and were anemic, as defined by a screening hemoglobin < or = 11 g/dL. Darbepoetin alfa was administered at a starting dosage of 3 microg/kg every 2 weeks for up to eight doses (16 weeks) in an open-label, noncomparative setting. A total of 194 oncology practices contributed 1,173 subjects to this interim analysis. The mean increase in hemoglobin was 1.7 g/dL (95% CI: 1.6, 1.8) to last value on study (intent-to-treat analysis) and 2.1 g/dL (95% CI: 1.9, 2.2) for those patients receiving the full 16 weeks of therapy. The Kaplan-Meier estimate of the proportion of subjects with a hematopoietic response (increase in hemoglobin > or = 2 g/dL and/or hemoglobin value > or = 12 g/dL) was 84% (95% CI:81,86). Subjects in the lower baseline hemoglobin category (< 10 g/dL) tended to have a greater hemoglobin response during treatment. The Functional Assessment of Cancer Therapy-Fatigue (FACT-Fatigue) subscale score increased by a mean of 6.8 points (26%) during the study, and improvements in fatigue paralleled the increases observed in hemoglobin. Study treatment-related toxicity was minimal, with the most common event being injection-site pain, seen in 2% of subjects. Experience to date with an every-2-week regimen of darbepoetin alfa indicated efficacy comparable to historical experience with weekly and 3-times-weekly regimens of epoetin alfa in treating chemotherapy-induced anemia in cancer subjects. PMID- 15352657 TI - Palliative care and the ethics of research: Medicare, hospice, and phase I trials. PMID- 15352658 TI - Involuntary weight loss: the forgotten vital sign. PMID- 15352659 TI - Suggestions for patients on how to talk with children about a parent's cancer. PMID- 15352660 TI - The changing impact of lifestyle and health. PMID- 15352661 TI - Trichofolliculoma with incidental acantholytic dyskeratosis. PMID- 15352662 TI - Tree stand falls: a persistent cause of sports injury. AB - OBJECTIVE: Tree stand falls are a well-known cause of hunting-related injury. Spine and brain injuries associated with these falls result in a significant incidence of permanent disability. Prior studies indicate that hunting tree stand injuries are largely preventable with the proper use of safety belts; however, compliance with safety belt use is variable. The purposes of this study were to determine 1) current compliance with safety belt use, 2) alterations in the spectrum of injury, and 3) causes of the falls. METHODS: From January 1996 to October 2001, 51 tree stand-related injuries referred to either of two regional trauma centers or their region's medical examiner's office were reviewed. Data had been recorded in each hospital's trauma registry, and the registries were searched for falls. Medical records were reviewed for additional data retrospectively, with an emphasis on determining the use of safety belts, and mechanisms contributing to the fall. RESULTS: Fifty-one cases of tree stand associated injuries were identified. These injuries all occurred in men, with a mean age of 42.6 years (range, 22-69 years). Alcohol use was present in 10% of patients and in two of the three deaths. The mean Injury Severity Score was 18.1 (range, 2-75). The most common injuries were spinal fractures (51% of series) and extremity fractures (41% of series). Closed head injuries were identified in 24% and lung injuries were identified in 22% of patients. Abdominal visceral injuries were present in 8% and genitourinary injuries were present in 4%. Three patients died. In addition to injury from the fall, a significant number (six patients [12%]) had additional morbidity from exposure. Only two patients reported the use of a safety belt (4% of series). There were no cases of gunshot wounds in this review, either self-inflicted or hunter-related. The chief reasons reported for these falls were errors in placement that resulted in structural failure of the stand, or errors made while climbing into or out of the stand (50% of falls). CONCLUSION: Devastating spine and brain injuries continue to occur after falls from tree stands during recreational hunting when safety belts are not used. Our results suggest a continuing need for the education of hunters concerning safe tree stand hunting practices, including proper methods of stand placement, assessment of tree branch strength, avoidance of fatigue and alcohol, anticipation of firearm recoil, and proper methods of stand entrance and exit. Trauma prevention programs directed toward heightened public awareness of these injuries during hunting season are still needed. PMID- 15352663 TI - Ulnar nerve injury associated with trampoline injuries. AB - OBJECTIVE: This study reports three cases of ulnar neuropathy after trampoline injuries in children. METHODS: A chart review was performed on children who sustained an ulnar nerve injury from a trampoline accident. In all cases, surgical intervention was required. RESULTS: Injuries included upper-extremity fractures in two cases and an upper-extremity laceration in one case. All cases required surgical exploration with internal neurolysis and ulnar nerve transposition. Nerve grafts were used in two cases and an additional nerve transfer was used in one case. All patients had return of intrinsic hand function and sensation after surgery. CONCLUSION: Children should be followed for evolution of ulnar nerve neuropathy after upper-extremity injury with consideration for electrical studies and surgical exploration if there is no improvement after 3 months. PMID- 15352664 TI - Does intraoperative electromyographic monitoring in lumbar microdiscectomy correlate with postoperative pain? AB - OBJECTIVES: Our objective was to correlate the findings of intraoperative electromyographic (EMG) monitoring with immediate postoperative pain in patients undergoing lumbar microdiscectomy. METHODS: A total of 112 patients undergoing de novo lumbar microdiscectomy were prospectively randomized into a control group (n = 45) and a study group (n = 67) in which intraoperative EMG monitoring was used. Postoperative pain and postoperative narcotic consumption were recorded for each patient. RESULTS: The presence or absence of EMG monitoring did not influence the level of reported pain in any anatomic area. In the monitored group, the degree of recorded nerve root irritation did not correlate with reported pain or postoperative narcotic consumption. The level of back pain was found to be significantly higher than the level of hip and calf pain (P < 0.0001). CONCLUSIONS: In our study no correlation was found between intraoperative EMG findings and immediate postoperative pain. PMID- 15352665 TI - Emergency department volume and acuity as factors in patients leaving without treatment. AB - OBJECTIVES: Patients who leave emergency departments (EDs) without treatment represent potential personal and hospital liability. Many department-dependent factors have been linked to patients who leave without treatment (LWT) in high volume EDs. The authors studied how department volume and acuity influence LWTs in a small, low-volume, university-affiliated ED. METHODS: Through retrospective ED census review, LWTs, department volume, and department acuity were recorded for 12-hour shifts over 1 year. Department acuity is defined as patients requiring resuscitation or admission. RESULTS: Over a 12-month period, 629 of 18,664 patients left the ED. When shift volume exceeded 25 patients, there were significantly more LWTs. When department acuity exceeded four patients per shift, there were also significantly greater numbers of LWTs. More than half of all shifts exceeded one of these thresholds. CONCLUSIONS: Thresholds of 25 patients and an acuity of five patients per shift were associated with significant increases in LWTs, suggesting possible per-physician maximum patient loads before an increased risk of LWT patients. PMID- 15352666 TI - Introduction. Sports medicine. PMID- 15352667 TI - Injuries to the foot and ankle in athletes. PMID- 15352668 TI - Articular cartilage injuries in the athlete's knee: current concepts in diagnosis and treatment. PMID- 15352669 TI - Injuries to the shoulder in athletes. PMID- 15352670 TI - Anterior cruciate ligament rupture. PMID- 15352671 TI - The patellofemoral joint. PMID- 15352672 TI - Fractures of the hand and carpal navicular bone in athletes. PMID- 15352673 TI - Neurapraxia, "stingers," and spinal stenosis in athletes. PMID- 15352674 TI - Trichofolliculoma with incidental focal acantholytic dyskeratosis. AB - Incidental focal acantholytic dyskeratosis has been described in a variety of cutaneous lesions, including benign and malignant epithelial lesions, fibrohistiocytic lesions, inflammatory lesions, and melanocytic lesions. It has also been observed in follicular lesions such as comedones and ruptured follicles. We report the case of a 47-year-old man with a firm, flesh-colored 2 mm pruritic papule in the sun-exposed area above the left eyebrow. An excisional biopsy was performed, the tissue was processed, and the hematoxylin and eosin slides were evaluated. Microscopic examination showed a dilated cystic cavity filled with keratinous debris and scattered fragments of hair. Smaller secondary follicular structures branched from the primary cyst's walls into the adjacent dermis. Hyperkeratosis, acantholysis, dyskeratosis, and suprabasilar clefts were also focally present. Correlation of the lesion's clinical morphology and microscopic features established a diagnosis of trichofolliculoma with incidental focal acantholytic dyskeratosis. Hence, trichofolliculoma can be added to the list of follicular lesions in which focal acantholytic dyskeratosis may be observed as an incidental microscopic change. PMID- 15352675 TI - Biceps pain as the presenting symptom of Parkinson disease: effective treatment with L-dopa. AB - Parkinson disease can present remarkably as shoulder pains that may long precede the clinical diagnosis. A case is presented in which, during a 2-year period, both lower biceps brachii became exquisitely painful, with both the pain and the "frozen shoulders" responding promptly to the initiation of L-dopa therapy. PMID- 15352676 TI - The femoral gateway. AB - A patient who abused cocaine intravenously had an abscess in the groin, which healed to form a sinus tract leading directly to the wall of the femoral vein. She used this "gateway" as her primary means of venous access for the next 5 years. It was both "painless and brainless" and led to femoral vein thrombosis and sepsis. This is the second reported case of such a unique means of venous access. PMID- 15352677 TI - The Greenfield filter as a potential hazard to the operating surgeon. AB - The Greenfield filter and similar devices are placed in the vena cava of patients who have contraindications to anticoagulation, but who would otherwise be at risk of pulmonary embolism without such anticoagulation. Injury to a health care worker from one of these devices has been reported in the case of a pathologist performing a necropsy. To the author's knowledge, injury or near-injury to a member of the surgical team during operation has not been reported in the literature. The authors report on a near-miss injury from such a device to the surgical first assistant during the performance of a pancreaticoduodenectomy. Simple guidelines to avoid similar injuries are provided. PMID- 15352678 TI - Marijuana-induced transient global amnesia. AB - A 6-year-old boy accidentally became intoxicated with marijuana secondary to ingesting cookies laced with marijuana. He presented with retentive memory deficit of sudden onset that was later diagnosed as transient global amnesia. Transient global amnesia as a result of marijuana intoxication is an extremely rare event. PMID- 15352679 TI - Multifocal skeletal tuberculosis. AB - Multifocal skeletal tuberculosis is defined as osteoarticular lesions that occur simultaneously at two or more locations. We present radiologic findings in two cases of multifocal osteoarticular tuberculosis. Differential diagnoses of such lesions, based on different radiologic modalities, can include metastatic disease, eosinophilic granuloma, or lymphoma. Since tuberculosis can be present in multiple sites, especially in patients from areas where tuberculosis is endemic, it is essential to avoid a delay in diagnosis. PMID- 15352680 TI - Disseminated Candida tropicalis in a patient with chronic mucocutaneous candidiasis. AB - Chronic mucocutaneous candidiasis is a heterogeneous group of immunodeficiencies associated with persistent candidal infections. Patients with chronic mucocutaneous candidiasis are rarely associated with systemic infections caused by other fungi, but almost never by Candida. The authors report a case of a 16 year-old with chronic mucocutaneous candidiasis who developed a fungemia with Candida tropicalis. PMID- 15352681 TI - Zolpidem for postanoxic spasticity. AB - A 28-year-old male sustained anoxic brain damage following aborted cardiac arrest, and subsequently developed severe muscular rigidity and spasticity involving all extremities. The spasticity was refractory to the standard regimens used for spastic hypertonia. Zolpidem dramatically inhibited muscular rigidity, spasticity, and dystonic posturing in a dose-dependent manner, resulting in a sustained improvement of his global performance over four years. The authors postulate a central mechanism of action by selective inhibition of GABAergic inhibitory neurons, and suggest a controlled clinical study to investigate the potential efficacy of zolpidem in relieving spasticity related to postanoxic brain injury. PMID- 15352682 TI - Physician values and medical decisions. PMID- 15352683 TI - Use of cryotherapy to isolate infected extremities during amputation. PMID- 15352684 TI - Black strap molasses for the treatment of inflammatory bowel disease-associated anemia. PMID- 15352685 TI - The patient's page. PMID- 15352686 TI - Hepatitis C virus transmission in the prison/inmate population. PMID- 15352688 TI - Expediting E-prescribing, getting Medicare beneficiaries on the Web. PMID- 15352687 TI - Perspectives. HSAs set for boom, but potential as personal savings vehicle is low. PMID- 15352689 TI - Information sources and level of knowledge of contraception issues among Greek women and men in the reproductive age: a country-wide survey. AB - OBJECTIVE: Good reproductive health depends to a great extent on how well informed people are on contraception issues. Contrary to the situation existing in many European countries, in Greece there has been a lack of studies investigating how well informed Greek women and men are. Also, the sources on which they draw are equally important. The aim of this study was to examine the knowledge of Greek women and men on issues of contraception, with the ultimate goal of identifying which groups should be the focus of prevention planning programs. METHOD: A country-wide survey was conducted through a self-administered questionnaire. The sample, numbering 1500 people, was representative of the Greek population aged 16-45 years. RESULTS: A key finding was that only a small percentage of the respondents were able to answer correctly 50% or more of the questions on knowledge of basic contraceptive issues (30.6% of women and 14.7% of men). Regarding sources of information, media and friends were reported as the primary sources of information for men of all ages and young women. The gynecologist becomes a significant source of consultation for women only after the age of 25 years. Even then, only 4% of women visiting a gynecologist do so to obtain information on prevention of pregnancy. The majority of women asking professional advice have already experienced an unwanted pregnancy. CONCLUSION: The need for health education is apparent, especially for young people. PMID- 15352690 TI - Frequent low-dose misoprostol for termination of second-trimester pregnancy. AB - OBJECTIVES: To determine the efficacy of an application regimen of low-dose frequent misoprostol for second-trimester pregnancy termination. METHODS: A total of 250 women between 12 and 20 weeks of gestation who were scheduled for second trimester pregnancy termination received 200 microg vaginal misoprostol followed by 100 microg oral misoprostol every 2 h until expulsion of the fetus. Mechanical cervical dilatation with a 16-French Foley balloon catheter was performed if no cervical dilatation was observed after 24 h. The main outcome measures were the delivery rate within 24 h and the factors influencing the interval between the onset of induction and abortion. Secondary outcome measures were the side-effects of the regimen and the total misoprostol dose required. RESULTS: With application of this protocol, 245 women (98%) delivered within 24 h of induction. The mean (+/-standard deviation) misoprostol dose used was 728+/-297 microg (200-2100 microg). Cox regression analysis revealed that vaginal spotting or nulliparity do not effect the induction-abortion time. On the other hand, using this regimen induction to abortion time tends to be longer in the presence of live fetuses (odds ratio (OR) = 0.45; confidence interval (CI) =0.2-0.8; p=0.008) and pregnancies with gestational age > 16 weeks (OR= 0.59; CI = 0.4-0.8; p= 0.003) when compared with cases of in utero death and pregnancies with a gestational age of 12-13 weeks, respectively. Twenty-seven women (10.8%) experienced one or more side-effects attributable to misoprostol. CONCLUSION: The 100-microg oral misoprostol every 2 h following 200 microg vaginal misoprostol is a highly effective protocol for inducing abortion at 12-20 weeks of pregnancy. Cases with live fetuses or pregnancies with older gestational age (> 16 weeks) deliver in a longer time period. PMID- 15352691 TI - The relationship between women's satisfaction with prenatal care service and the characteristics of the pregnant women and the service. AB - OBJECTIVE: The purpose of this descriptive correlational study was to investigate the relationship between women's satisfaction with prenatal care services and the demographic characteristics of the women and the service's features. METHODS: The population studied consisted of Turkish women living in Erzurum, Turkey, who had delivered their infants and were still hospitalized. The women had received prenatal care, had no complications during pregnancy, carried their pregnancies to term, and were considered to have had normal deliveries. The sample size was determined as 367 women, and 350 women responded to the questionnaire. Question items consisted of women's satisfaction with prenatal care services, the demographics characteristics of the pregnant women, and the characteristics of prenatal care. The questionnaire was given to the women in the birthing house. The researchers collected data using an interview method between 1 January and 30 April, 2002. RESULTS: In terms of the mean item score and total scale, the women were 'somewhat satisfied' with the prenatal care service. The women were more satisfied with health-care information and the 'cost of the service' dimension of the scale. There was an association between women's satisfaction with prenatal care services and the characteristics of both prenatal care and those of the pregnant women. CONCLUSION: The sample of women in this study reflects only the study population in this area of Turkey. The findings of the study should be limited to this population. The scale would be useful for further studies investigating how prenatal care providers can better identify women's satisfaction with prenatal care service. A satisfaction measurement scale of prenatal care services would be helpful as an interview form that could identify the satisfaction scale and rate the scale's importance in the health-care provider's office, hospital, during a home visitation, or a primary health-care center, private clinic or maternity and child health center. PMID- 15352692 TI - Attitudes of Israeli gynecologists towards the levonorgestrel-releasing intrauterine system: a questionnaire survey. AB - OBJECTIVES: The aim of the study was to examine the current attitudes and practices of gynecologists regarding the use of the levonorgestrel-releasing intrauterine system (LNG-IUS, Mirena, Schering AG, Germany) as a contraceptive device and for the treatment of menorrhagia in the premenopausal period. In addition, physician knowledge of patient satisfaction with the device was evaluated. METHODS: A written, self-administered questionnaire was given to 300 gynecologists actively employed in gynecological departments and outpatient clinics in urban centers in Israel. The items covered the indications for which the LNG-IUS was prescribed, the estimated rate of use of the LNG-IUS compared to copper intrauterine devices (IUDs), patient satisfaction with the LNG-IUS and previous knowledge about it, and demographic characteristics of the population using IUDs. All forms were completed on an anonymous basis. RESULTS: The questionnaire was returned, properly completed, by 298 physicians, yielding a response rate of 99.3%. Almost one-half of the physicians (n = 147, 49.3%) used the LNG-IUS only as a contraceptive device, 17.4% only as treatment for menorrhagia, and 33.2% for both indications. The vast majority of gynecologists (82.2%) reported a patient satisfaction rate of 70% or more. Among the gynecologists using the LNG-IUS as treatment for menorrhagia, the estimated overall patient satisfaction rate was more than 50%. Although most physicians expressed greater patient satisfaction with the LNG-IUS than with copper releasing IUDs (p < 0.004), the majority of gynecologists still used the copper releasing IUDs more frequently. Most considered the LNG-IUS as an appropriate method for endometrial protection in postmenopausal women using estrogen replacement therapy. CONCLUSIONS: Physicians should be made aware of the beneficial health effects of the LNG-IUS, particularly in premenopausal women with menorrhagia and for endometrial protection during hormone replacement. PMID- 15352693 TI - Attitudes towards male condom use in two different populations in Thrace, Greece. AB - OBJECTIVE: To outline Thracian women's disposition and attitude towards the male condom. METHODS: Representatives of the two major religious subgroups in Thrace (127 Christian Orthodox and 120 Muslim women) were studied. All respondents were of reproductive age (from 17 to 39 years) and were encouraged to answer a specific questionnaire. Women expressed their opinions about their mood while using it, its safety and similarity to natural contraception, and its contribution to the prevention of carcinogenesis and sexually transmitted diseases (STDs). They were also questioned about the ease of accessibility to supplies, the convenience experienced in buying it, its cost, and whether they thought it was necessary to use one in every act of intercourse as a contraceptive method. The statistical software package used was the Sigma Stat 2.0. RESULTS: The majority of the respondents in both subgroups had a positive opinion about the condom's contribution to the prevention of acquired immune deficiency syndrome (AIDS) and STDs. Christians were better informed about the condom's contribution to the prevention of carcinogenesis, while the majority of Muslims did not feel convinced about the condom's contraceptive efficacy. CONCLUSIONS: The study results reveal a significant difference in the way that Christian and Muslim women in Thrace consider the impact of condom use on sexual behavior and reproductive health care. PMID- 15352694 TI - Experience with Implanon in a northeast London family planning clinic. AB - AIM: The primary objective of the study was to evaluate the continuation rates of a relatively new long-acting method of contraception, Implanon. The secondary objective was to study discontinuations related to bleeding problems and their management. METHOD: A retrospective review of records of women fitted with Implanon was undertaken during February 2000--January 2003. RESULTS: Results from the secondary objective will be the subject of a separate communication. Of the 147 implants fitted, 132 records could be retrieved; of these, 97% of the women had pre-insertion counselling by the clinician. The most common indications for Implanon usage were choice of a long-acting method, unhappy experience with other contraceptive methods and suboptimal compliance with contraceptive pills and injectables. The median age of fitting was 25 years and 36% of the women were nulliparous. No problems were experienced with fitting or removal of implants. Twenty (15%) women were lost to follow-up. Twenty-two implants were removed by the end of the study period, with 12 (60%) removals attributed to prolonged heavy/light bleeding. There were no known pregnancies during the study. Given that 15% of the women could not be followed up or contacted, the assumed lifetimes of Implanon using the Kaplan-Meier method are 0.90 (95% confidence interval 0.82-0.95) at 12 months, 0.80 (0.67-0.88) at 24 months and 0.75 (0.58 0.85) at 35 months. The confirmed lifetimes are 0.84 (0.71-0.91) at 12 months, 0.63 (0.42-0.78) at 24 months and 0.53 (0.28-0.73) at 35 months. CONCLUSION: Although not free of side-effects, Implanon can be a good choice for women who are properly informed and counselled and seek long-term 'forgettable' contraception. PMID- 15352695 TI - Family planning and psychosocial support for infertile couples. AB - AIMS: The study was set up to analyze the psychological/emotional needs of women who undergo treatment for in vitro fertilization (IVF) and to emphasize the importance of the psychosocial support that family planning centers can provide to them. METHOD: This was a cohort study with closed questions. A total of 235 infertile women participating in an IVF program were studied. The statistical analysis of the findings was conducted by the Statistical Package for Social Sciences. RESULTS: The psychosocial support and the scientific information provided to those women who participate in IVF programs are insufficient. In order to succeed in an IVF program, the women must be assisted by a doctor with much medical experience. The study shows that 59.3% of the women studied sought more medical information and another 32.5% sought emotional support. These percentages varied with reference to education, age and origin. These kinds of services must be provided by family planning centers. PMID- 15352696 TI - [Intervertebral disk prolapse: how long is conservative treatment adequate?]. PMID- 15352697 TI - [Minimally invasive procedures of the intervertebral disk. They don't always produce what they promise]. PMID- 15352698 TI - [Don't allow yourself to be misled by overanxious parents. Asthma children should not be excused from school sports]. PMID- 15352699 TI - [New diabetes therapy in clinical trial: will need adapted insulin supply be available soon? (interview by Dr. Judith Neumaier)]. PMID- 15352700 TI - [Homeopathy of classical medicine: your prescription for cough, common cold, hoarseness?]. PMID- 15352701 TI - [Current controversies in endocrinology: obesity epidemic, hormone replacement therapy, latent thyroid gland disorders]. PMID- 15352702 TI - [Hormone replacement therapy and the risk of developing breast cancer]. AB - The Million Women Study (MWS) and the Women's Health Initiative (WHI) Study reveal an increased breast cancer risk and a higher relative risk for cardiovascular diseases in women on hormone replacement therapy (HRT) in comparison with control groups. For this reason, the WHI study was terminated prematurely. From the point of view of the internist, it would appear that, on the basis of the results of these studies, prophylactic treatment with estrogens and progesterone is no longer indicated. It must, however, be noted that numerous aspects are still unclear. This therefore means that in patients with severe menopausal symptoms giving rise to a high level of distress, HRT continues to be justified over the short term after a current cardiovascular risk or relevant tumor disease has been excluded. PMID- 15352703 TI - [Thyroid gland malfunction]. AB - Uncharacteristic clinical symptoms such as cardiac arrhythmias, restlessness, changes in weight or hair loss may signify hyperthyroidism, whereas impairment of physical or mental vigilance may be a manifestation of hypothyroidism. In many cases the diagnosis can be established only on the basis of laboratory investigations, and, on account of the difficulty in establishing a differential diagnosis, these should not be stinted. However, when it comes to interpreting findings, consideration must be given to drug interference, such diseases as the nephrotic syndrome, serious general diseases or pregnancy. In the event of pathological laboratory findings, ultrasonography and scintigraphy are helpful further diagnostic measures. Antihyperthyroid drugs, surgery or radioiodine therapy are the leading therapeutic options for hyperthyroidism. As in the case of hypothyroidism treated with L-thyroxine, the primary target parameter is titration of TSH into the normal range. PMID- 15352704 TI - [Minimally invasive surgery in endocrinology--potentials and limitations]. PMID- 15352705 TI - [Electromagnetic pollution (electrosmog)--potential hazards of our electromagnetic future]. AB - The term electromagnetic environment encompasses the totality of all electric, magnetic and electromagnetic fields generated by natural and technical sources. A differentiation is made between low- and high-frequency electromagnetic fields. Typical sources of the former are domestic electricity Exposure to the latter is, for example, associated with the sue of mobile telephones. Studies on the health related effects of electromagnetic fields are available in particular for the low frequency range, based on an appropriate estimation of exposure. A number of these studies reveal an association between exposure to this type of electromagnetic fields and the occurrence of infantile leukemia in the highest exposure category. For high-frequency electromagnetic fields the number of epidemiological studies is limited. An increased risk of an accident occurring through the use of a cellular phone while driving has consistently been shown. Against the background of our limited knowledge about possible adverse effects of exposure to mobile phone transmitters, and the inability of the public to influence such exposure, transparency in the communication of the risks involved is of great importance. PMID- 15352706 TI - [Hypertensive crisis/hypertensive emergency: how are they defined and what's to be done?]. PMID- 15352707 TI - [When Ulla Schmidt finally signs: bureaucracy insanity of DMPs will finally dissipate]. PMID- 15352708 TI - [DMP requires more education than the family physician can afford. How colleagues solve the problem]. PMID- 15352709 TI - [New evaluation by the EU approval committee. Benefits of cox-2 inhibitors outweigh possible risks (interview by Dr. Thomas Meissner)]. PMID- 15352710 TI - [10 minute consultation: vertigo in the elderly]. PMID- 15352711 TI - Advantages and limitations of the internet in scientific literature searches. PMID- 15352712 TI - Pesticide analysis by capillary electrophoresis. AB - In this work, a critical and updated revision of the current situation of the analysis of pesticides by Capillary Electrophoresis (CE) is presented. The review has been written in two main sections. The first one presents a thorough revision of the various offline and on-line sample preconcentration procedures that have been used in conjunction with CE to analyze these compounds. The second part reviews the various detection strategies (i.e., UV, LIF, MS, and electrochemical) and CE modes that have been applied to the analysis of pesticides. Future trends that can be expected from this hot research area are also discussed. PMID- 15352713 TI - Rapid analysis of triterpenic acids by liquid chromatography using porous graphitic carbon and evaporative light scattering detection. AB - An original system which uses Porous Graphitic Carbon as support and a mixture of organic solvents as mobile phase is proposed for the analysis of triterpenic acids by liquid chromatography. The separation of betulinic acid, ursolic acid, oleanolic acid, and 18alpha- and 18beta-glycyrrhetinic acids was carried out within a short time and monitored by evaporative light scattering detection as universal detection method. Molecular modelling studies show that the main contribution to the selectivity comes from the electrostatic interaction characterised by the dipole moment of the products. PMID- 15352714 TI - Separation and characterisation of sphingoceramides by high-performance liquid chromatography-electrospray ionisation mass spectrometry. AB - We developed a simple and reliable analytical method for the quantification and the characterization of ceramides extracted from biological samples by high performance liquid chromatography (HPLC) coupled to electrospray ionisation tandem mass spectrometry (ESI/MS/MS). The chromatographic separation of analytes was carried out in a RP8 column, eluting with a methanol-water mixture in gradient elution mode. The separated lipids were detected by total ion monitoring and characterised by MS/MS spectra; quantitative analysis was performed by integrating the extracted ion peaks obtained in the negative ion mode. Good repeatability was obtained for retention time (0.3-2%), peak area ratio (A(S)/A(IS), 2-8%), as well as limit of detection (LOD, 5-26 pg) and quantification (LOQ, 13-53 pg). The method was validated for the analysis of N palmitoyl-D-erythro-sphingosine (Cer16), N-stearoyl-D-erythro-sphingosine (Cer18), N-tetracosanoyl-D-erythro-sphingosine (N24:0, lignoceric ceramide, Cer24:0), and N-tetracos-15'-enoyl-D-erythro-sphingosine (N24:1, nervonic ceramide, Cer24:1), giving good results. Lipid mixtures, extracted from skin and epidermal cells, were analysed for their content of the studied ceramides. PMID- 15352715 TI - Separation of racemic 2,4-dinitrophenyl amino acids on zirconia-immobilized quinine carbamate in reversed-phase liquid chromatography. AB - Zirconia is known to be one of the best chromatographic support materials due to its excellent chemical, thermal, and mechanical stability. A quinine carbamate coated zirconia was prepared as a chiral stationary phase for separation of enantiomers of DNP-amino acids in reversed-phase liquid chromatography. Retention and enantioselectivity of this phase were compared to those for quinine carbamate bonded onto silica. Most amino acids studied were separated on the quinine carbamate-zirconia CSP although retention was longer and chiral selectivity was somewhat lower than on the corresponding silica CSP. Increased retention and decreased selectivity are probably due to strong non-enantioselective Lewis acid base interactions between the amino acid molecule and the residual Lewis acid sites on the zirconia surface. PMID- 15352716 TI - Study of the mechanism of enantioseparation part VI: thermodynamic study of HPLC separation of some enantiomers of phenylcarbamic acid derivatives on a (S,S) Whelk-O 1 column. AB - The enantiomers of thirteen 2-, 3-, and 4-alkoxyphenylcarbamic acid 2-methoxy-1 [(4-methylpiperazino)methyl]ethyl ester were separated on a (S,S) Whelk-O 1 CSP column isothermally in the range of 0-50 degrees C at 10 degrees C increments, using methanol/ water (90/10, v/v) containing 17.5 mmol L(-1) acetic acid and 14.36 mmol L(-1) triethylamine as a mobile phase. The dependence of the natural logarithms of retention and selectivity factors (In k, In alpha, respectively) on the inverse of temperature, 1/T, was used to determine thermodynamic data of enantiomers of alkoxysubstituted phenylcarbamic acid 2-methoxy-1-[(4 methylpiperazino) methyl]ethyl esters. Enthalpyentropy compensation plots showed that all of the compounds in this study separate via the same enthalpy-driven chiral recognition mechanism. PMID- 15352718 TI - Multi-criteria decision making in micellar liquid chromatographic separation of chlorophenols. AB - Simultaneous optimization of separation quality and analysis time of the micellar liquid chromatography of nine chlorophenol isomers was investigated. The effect on retention of three experimental parameters was studied using multivariate analysis. The factors studied were the concentration of sodium dodecyl sulfate, propanol content, and pH of the mobile phase. The experiments were performed according to the face-centered cube central composite design and the inverse form of the experimental retention times of analytes was fitted to polynomial models. The results of the analysis of variance showed that the models obtained explain over 99% of the variance observed in the chromatograms. The good predictive ability of the models was verified by high correlation coefficient (R2 > 0.99) and F ratio values for the plots of predicted cross-validated versus experimental retention times. The study showed that the use of the Pareto-Optimality method, an approach from multi-criteria decision making, allows selection of the best possible combinations of separation quality and analysis time in micellar liquid chromatography of chlorophenols. PMID- 15352717 TI - Optimization of micellar liquid chromatographic separation of polycyclic aromatic hydrocarbons with the addition of second organic additive. AB - The micellar liquid chromatographic (MLC) separations of polycyclic aromatic hydrocarbons (PAHs) were optimized for three micellar systems, cetyltrimethylammonium chloride (CTAC), dodecyltrimethylammonium chloride (DTAC), and sodium dodecylsulfate (SDS), with 1-pentanol as the only organic additive. A difference in the separation was observed between CTAC and SDS/DTAC. Under each optimized separation conditions, CTAC-modified mobile phase provides the least desirable separation, which is attributed to its longer carbon tail (C16 vs. C12). In addition to 1-pentanol, the main organic additive, a second organic additive (3% 1-propanol) in the micelle-modified mobile phase was found to enhance the resolution of PAH chromatographic peaks. However, the extent of the enhancement varies for the different micellar systems, with the greatest resolution improvement seen for CTAC, and little effect for shorter-tail SDS and DTAC. This study shows the potential use of second organic additive (1-propanol), to the main nonpolar additive (1-pentanol), in facilitating the MLC separation of larger nonpolar compounds. PMID- 15352719 TI - Investigation of the factors that induce analyte peak splitting in capillary electrophoresis. AB - Peak splitting has a detrimental effect on analyses by capillary electrophoresis. Many papers have reported it and several mechanisms have been proposed to explain the phenomenon. We investigated the electrophoretic behavior of an amphoteric analyte, levodopa, in phosphate buffer and observed a peak splitting phenomenon at moderate sample concentrations and under general analytical conditions, even without organic solvent. The dependence of effective mobility on pH was taken into account and pKa values of 2.30, 8.11, and 9.92 were obtained for levodopa. Then, we constructed pH-dependent distribution diagrams of levodopa and phosphate species present in aqueous solution and proposed that the most relevant factors contributing to peak splitting are the presence of ionizable groups in the analyte molecule and the occurrence of ionization, yielding charged species which interacted with buffer electrolyte species in a definite pH range to form complexes. This result is different from those presented in the literature and broadens our understanding of amphoteric analyte peak splitting. PMID- 15352720 TI - A preliminary approach to the separation of Leishmania cell-surface antigens. AB - The purpose of the current study was to characterize Leishmania cell-surface antigens by two different methods established for the purification of glycoproteins and proteins, and to point out a useful approach to define their size and mass heterogeneity. L. tropica parasites were initially isolated from patients with active cutaneous leishmaniasis and were then cultured in vitro. The parasite-cell layer was solubilised with 6 M guanidinium chloride (GuHCl) and subsequently prepared for the purification procedure. The methods used in this work were gel filtration chromatography and isopycnic density-gradient centrifugation. Because of the presence of a substantial amount of non-specific proteins in the culture medium, these methods were not effective alone in distinguishing these antigens. However, a good idea of their N-glycosylated structures could be obtained by using Periodic acid-Schiffs (PAS) and Con A lectin, and also size and mass heterogeneity. A combination of these methods effected a clear separation of the antigens. Amino acid analysis of the purified antigens was performed to positively identify them as well-known Leishmania cell surface antigen gene products. The results confirmed the presence of more than one cell-surface antigen on the Leishmania parasite and the combination of gel chromatography and density-gradient centrifugation could be useful for their isolation. PMID- 15352721 TI - Dynamic solid phase microextraction for sampling of airborne sarin with gas chromatography-mass spectrometry for rapid field detection and quantification. AB - A portable dynamic air sampler and solid phase microextraction were used to simultaneously detect, identify, and quantify airborne sarin with immediate analysis of samples using a field portable gas chromatography-mass spectrometry system. A mathematical model was used with knowledge of the mass of sarin trapped, linear air velocity past the exposed sampling fiber, and sample duration allowing calculation of concentration estimates. For organizations with suitable field portable instrumentation, these methods are potentially useful for rapid onsite detection and quantification of high concern analytes, either through direct environmental sampling or through sampling of air collected in bags. PMID- 15352722 TI - Chiral separation of omeprazole and several related benzimidazoles using supercritical fluid chromatography. AB - A study of the enantiomeric separation of omeprazole and several related benzimidazoles, using supercritical fluid chromatography (SFC), on the amylose based column Chiralpak AD is presented in this work. The effect of the organic modifier as well as temperature on the retention and enantioresolution was investigated. Alcohol-type modifiers provided the best results, allowing the enantiomeric separation of all the compounds studied with resolutions that were in most cases higher than 2, and analysis times lower than 10 minutes. An investigation of the temperature effect revealed that the isoelution temperature was below the working temperature range in only two cases, and hence it was better to work at the highest temperature permitted. PMID- 15352723 TI - Rapid and sensitive determination of morphine in street opium samples by thermal desorption gas chromatography using a microfurnace pyrolyzer. AB - Thermal desorption of the alkaloids in opium samples at 300 degrees C using a vertical microfurnace pyrolyzer was followed by their on-line gas chromatographic (GC) analysis on a large-bore glass capillary column. This method permitted rapid and sensitive determination of the content of the main alkaloid, morphine, in the small (ca. 100 microg) opium samples with a relative standard deviation within 4% for 5 runs. The observed morphine contents of about 12 to 15 w/w% in the given opium samples were in fairly good agreement with those estimated by a conventional GC-MS method. PMID- 15352724 TI - Resolution of triacylglycerol positional isomers by reversed-phase high performance liquid chromatography. AB - The ability of reversed-phase high-performance liquid chromatography (RP-HPLC) to separate some positionally isomeric disaturated and monounsaturated triacylglycerols (TAGs) as intact species is demonstrated for the first time. Mobile phases of acetonitrile modified with methanol, ethanol, 2-propanol, 1 propanol, 1-butanol, acetone, or dichloromethane were tested for the separation of POP-PPO, PLP-PPL, PEP-PPE, and PDP-PPD (P-palmitic, O-oleic, L-linoleic, E eicosapentaenoic, D-docosahexaenoic acid residue) on a single RP-HPLC column. The resolution improved with increasing number of double bonds in the acyl residues. While POP and PPO were only partially resolved, PDP and PPD were fully separated with all tested mobile phases, except those containing methanol. Also separated were the four TAGs having the same equivalent carbon number (ECN = 42), PEP, PPE, PDP, and PPD, on a single RP-HPLC column with mobile phase acetonitrile-2 propanol (70:30, v/v) at 0.8 mL/min. In all cases the isomer with the unsaturated acyl residue in either 1- or 3-position was retained more strongly than the respective 2-isomer. PMID- 15352725 TI - Behavior of new complexes of tetrakis(4-methoxylphenyl)porphyrin with heavy rare earth elements in reversed-phase high performance liquid chromatography. AB - An HPLC method has been developed for the separation of new complexes of tetrakis(4-methoxylphenyl)porphyrin (TMOPP) with four heavy rare earth elements (RE = Y, Er, Tm, and Yb). The function of amine and acid in the mobile phase has been investigated and a reasonable explanation is presented. Successful separation of the RE-TMOPP-Cl complexes is accomplished in 10 min with a mobile phase consisting of methanol-water-acetic acid-triethanolamine. The detection limits (S/N= 3) for the four complexes are 0.01 microg/mL. This method is rapid, sensitive, and simple. PMID- 15352726 TI - Enrichment of the glycoalkaloids alpha-solanine and alpha-chaconine from potato juice by adsorptive bubble separation using a pH gradient. AB - For the first time, the solanidine alkaloids alpha-solanine and alpha-chaconine could be quantitatively enriched from potato juice by Adsorptive Bubble Separation (ABS) with a pH gradient. The enrichment into the foam was influenced by the pH value, bubble size, and gas flow rate. The efficiency was highest on using diluted samples with a concentration between 2 and 6 mg L(-1) of the alkaloids at pH 6.0. The experiments with a standard solution of each alkaloid confirmed that these substances can be quantitatively enriched into the 'spumat' without surface active potato proteins. The transfer into the foam fraction under these conditions was similar to that from the aqueous potato extract. PMID- 15352727 TI - Is there no controversy in chromatography? Review of the 27th International Symposium on Capillary Chromatography Riva del Garda, Italy, May 31-June 4, 2004. PMID- 15352728 TI - Enhancing collaborative tendencies: extending the single identity model for youth conflict education. AB - A conflict intervention in postapartheid South Africa suggests new insights into encouraging an honest and engaging participation in the dialogue process. PMID- 15352729 TI - Seeds of peace: toward a common narrative. AB - In the woods of Maine, young people from conflict regions meet to learn about themselves, each other, and their potential to promote peace. PMID- 15352730 TI - Normalizing effective conflict management through academic curriculum integration: the example of workable peace. AB - An innovative program being implemented in U.S. school systems teaches youth to integrate the skills of conflict management beyond academics to their social and civic lives. PMID- 15352731 TI - After the fall: a conflict management program to foster open society. AB - A conflict management program developed to support the transition from a closed to open society in postcommunist eastern and central Europe focuses on the future leaders of this region: adolescents. PMID- 15352732 TI - Youth intervention for peace project: Burundi case study. AB - Youth in war-ridden Burundi embrace the skills of conflict management in their process to become positive participants in their country's political process. PMID- 15352733 TI - Challenging intolerance. AB - Social identity is multilayered and complex. Schools, communities, and nations can implement structures and approaches that help youth to understand one another and respect differences. PMID- 15352734 TI - The blue planet. PMID- 15352735 TI - Denizens of the deep. PMID- 15352736 TI - Swimming with giants. PMID- 15352737 TI - Feeling the sting. PMID- 15352738 TI - Don't go in the water. PMID- 15352739 TI - Exposure opportunity to substances of abuse. PMID- 15352740 TI - Curacao patients with coronary artery disease have a higher prevalence of the HFE C282Y mutation. AB - Epidemiological studies indicate a positive relation between iron status and coronary artery disease (CAD) risk The HFE C282Y allele is associated with increased iron status and higher CAD risk. We investigated whether HFE C282Ymight be a CAD risk factor in Curacao in a case-control study design. The patient group comprised 42 men and 10 women. Fifty-four men and 30 women without history of CAD served as age and gender matched controls. HFE C282Y genotypes were established using sequence-specific priming polymerase chain reaction. None of the investigated subjects were homozygous for HFE C282Y, whereas 5/52 (9.6%) CAD patients and 1/84 controls (1.2%) were heterozygous for HFE C282Y (p = 0.03). The HFE C282Y mutation was 8.8 fold (95% CI 1.001, 77.8; p = 0.049) more prevalent in CAD patients than in controls. The HFE C282Y allele frequency in Curacao is higher than that of African populations, but comparable with that of Jamaica. We conclude that Curacao CAD patients have somewhat higher frequency of HFE C282Y heterozygosity than controls, and that the HFE C282Y allele frequency in the Curacao population is higher than might be expected in persons of African descent. The consequences of HFE C282Y heterozygosity as CAD risk factor are as yet uncertain, since there is no proof that iron lowering reduces CAD risk. PMID- 15352741 TI - Serum malondialdehyde levels in patients infected with Plasmodium vivax. AB - Malondialdehyde, a lipid peroxide, may be used as an indicator of oxidative stress. This study investigated the oxidative stress hypothesis in patients infected with Plasmodium vivax. Serum malondialdehyde (MDA) concentration activity was measured in 87 patients. Scores were obtained for the patients and 60 seronegative healthy controls matched for age and gender. The difference between MDA levels of patients infected with Plasmodium vivax and the control group was statistically significant both for females (p < 0.001) and males (p < 0.001). In the patient and control group, no correlation was found between age and malondialdehyde levels (p > 0.05) in both females and males. In addition, no significant correlation could be found between MDA levels of both females and males for patients and control group (p > 0.05). MDA levels were increased in the patients infected with Plasmodium vivax. PMID- 15352742 TI - Incidence of cervical carcinoma in a high-risk, non-screened area results of a retrospective analysis on the Dutch Caribbean Antilles from 1983 to 1998. AB - Cancer of the uterine cervix is one of the most common malignancies worldwide. The average annual incidence of cervical cancer varies widely per geographical area. Some of the highest rates have been reported in Latin American and Caribbean countries. Previous research into the frequency of cervical carcinoma in Curacao, the main island of the Dutch Caribbean, has reported predominantly far-advanced stages of the disease. The objectives of this study were to determine: whether there are indications of changes in the incidence of cervical carcinoma, whether these changes are a result of increased clinical attention paid by general practioners (GPs) and gynaecologists, and whether less advanced stages of cervical cancer were found, indicating earlier diagnosis. Data on patients with cervical carcinoma, obtained from records kept in the Department of Pathological Anatomy at the St Elisabeth Hospital in Curacao were retrospectively analyzed for the period 1983 to 1998. To study the incidence rate over time, cumulative and incidence rates, standardized to the World Standard Population, were calculated and compared to the rates in other Latin American and Caribbean countries. The age-standardized incidence rate for Curacao was 14.3 per 100 000 women. The cumulative rate up to 74-years-of-age was 1.6%. Sub-division into the three groups showed a trend towards a decrease in the incidence and cumulative rates. A comparison of clinical staging International Federation of Obstetrics and Gynaecology (FIGO) showed a trend towards an increase in time of almost 48% to 59% in stage IA (ie less severe cases). A significant increase (p < 0.0001) was found in the numbers of smears performed in the complete period of 1983-1998, compared to a previously examined period of 1972-1982. The incidence of cervical cancer in Curacao appears to be decreasing, and is lower than in other areas in the region. The number of smears GPs performed over time increased However the incidence is still relatively high compared to western countries and cases are still presenting in relatively advanced stages of the disease. Although the introduction of a screening programme might not influence the actual incidence of cervical cancer dramatically, nonetheless it may contribute to earlier diagnosis of cervical carcinoma. PMID- 15352743 TI - HIV seroprevalence and risk factors in female substance abusers seeking rehabilitation in Trinidad and Tobago. AB - Crack cocaine use, especially among females, is associated with risky sexual behaviour. Data from 122 female substance abusers admitted to a rehabilitation centre in Trinidad and Tobago between 1996 and 2002 were retrospectively reviewed to determine Human Immunodeficiency Virus (HIV) seroprevalence and other associated risk factors. HIV seroprevalence was 19.7%, eight times higher than in the general population. Factors significantly associated with HIV infection were use of crack cocaine, low educational attainment and diagnosis of a sexually transmitted infection. Other factors associated with risky sexual behaviour were common--use of cocaine in combination with alcohol (65.6%), psychiatric illness (28.7%) and a history of sexual abuse (83%). Female substance abusers, especially female crack addicts are at high risk of acquiring and transmitting HIV. Rehabilitation programmes should include HIV prevention interventions which specifically target all risk factors including trauma counselling, skills development and screening for sexually transmitted infections. PMID- 15352745 TI - Survival and surface adherence ability of bacterial pathogens in oral liquid pharmaceuticals and their containers. AB - The survival and surface adherence ability of Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa in nutrient broth and in five oral liquid pharmaceuticals (nivaquine syrup, cough mixture, paracetamol elixir, cotrimoxazole and vitamin C) were investigated The bacteria grew more in nutrient broth than in the pharmaceuticals (p < 0. 001) and the recovery of stressed cells was enhanced when 3% Tween 80 was used as the recovery medium as against the use of normal saline (p < 0.01). The Gram-negative bacteria were more adapted to the pharmaceuticals than their Gram positive counterparts. Klebsiella pneumoniae and Ps. aeruginosa were recovered in large numbers from nivaquine and cotrimoxazole suspensions that did not support the growth of the other bacteria. The effect of bacterial growth on the physico chemical properties of the pharmaceuticals was also evaluated The properties were not altered significantly except for pH shifts of 0.3 to 1.1 caused by E. coli and S. aureus in paracetamol and vitamin C. Adherence capability was found to correlate with the survival ability of the bacteria. Populations on coupons were significantly higher when nutrient broth was used as the suspending medium compared with any of the pharmaceuticals (p < 0.01). Rubber and plastic coupons were significantly more accessible to the bacteria than glass coupon as revealed by the high population of bacteria recovered from their surfaces. PMID- 15352744 TI - Infections in neurosurgical patients admitted to the intensive care unit at the University Hospital of the West Indies. AB - Patients admitted to the Intensive Care Unit (ICU) are at risk of developing life threatening nosocomial infections, especially with organisms resistant to commonly used antibiotics. Neurosurgical patients are particularly vulnerable because of the serious nature of their illness, the frequency of associated trauma and the presence of invasive devices. Of 120 neurosurgical patients admitted to the ICU of the University Hospital of the West Indies (UHWI) between September 1995 and December 1999, the records of 73 patients were available for analysis. All had prophylactic antibiotics. Twenty-one of these 73 patients (28.8%) developed 22 infections after a mean of five days in the ICU: nine with chest infection, seven with urinary tract infection, four with central nervous system (CNS) infection and one each with wound and skin infection. This is an incidence of 11.6/1000 patient-days. The responsible organisms included Pseudomonas (7/21), Acinetobacter (3/21), E. coli 2/21, Enterobacter (2/21), and Klebsiella (2/21), and one each with Staphylococcus aureus, methicillin resistant Staphylococcus aureus, coagulase negative Staphylococcus, group D Streptococcus and bacteroides (1/21). Infection was significantly related to length of hospital stay, length of ICU stay, duration of intubation, duration of ventilation and the presence of diabetes mellitus. All patients who had surgery after ICU admission developed infection, seven with chest infection, two with urinary tract infection, two with CNS and one with skin infection. The three patients who were admitted with intracranial infections all developed other infections. Infected patients had a significantly longer hospital stay. Five patients died, none directly attributable to infection, while 55 (75.5%) made a good recovery. The problem of ICU infection may be expected to escalate with the increased use of intensive care, increasingly more complex surgical procedures and the growing problem of antibiotic resistance. Since infection is related to the length of ICU stay, earlier discharge of neurosurgical patients to an appropriately staffed high dependency unit is likely to result in reduction of the infection rate. Reinforcement of infection control strategies within the ICU may be expected to further minimize the infection rate. PMID- 15352746 TI - Cancer of the colon and rectum in a Jamaican population: diagnostic implications of the changing frequency and subsite distribution. AB - The aim of this study was to examine the clinical and pathological characteristics of colorectal cancer in Jamaica, to determine whether there was a change in the anatomic distribution and clinical presentation and to discuss the options for diagnosis and management. A comprehensive retrospective review of patients newly diagnosed with colorectal carcinoma was conducted at The University Hospital of the West Indies by reviewing both patient records and pathological data. These data were compared with previous reports of patients with colorectal cancer seen in Jamaica. One hundred and forty-seven patients were studied There were 85 females and 62 males with a female to male ratio of 1.37:1. The median age was 65.5 years (range 19 to 94 years). The predominant symptoms were abdominal pain in 91 patients, change in bowel habit in 77 patients and rectal bleeding in 74 patients. Sixty patients presented with weight loss and 28 with a rectal mass. The most common tumours were right-sided colonic cancers in 42 patients (28.5%) followed by sigmoid colon in 30 (20.4%) rectum in 34 (23.1%) and left and transverse colon accounting for 16 and 10 cases respectively. Most of the tumours were well or moderately differentiated adenocarcinomas. Only eight patients presented with Dukes' A disease, 50 with Dukes'B, 53 with Dukes'C disease and 34 with advanced disease. The findings showed that sigmoid and rectal tumours accounted for 43.5% of cancers. The colon/rectum ratio in this series was 3.3:1 indicating a significant proximal shift of colorectal cancers in this population in keeping with recent reports. The results of the current study suggest that the sub-site location of colorectal cancers seen is similar to that reported in high incidence countries such as the United States of America and parts of Europe but differs from the African continent which has a high proportion of rectal tumours. This right-sided preponderance also differs from previous studies in Jamaica, which report a higher incidence of rectal lesions The detection of early colorectal carcinoma will require screening at a stage when the disease is asymptomatic in order to improve the chance for cure. The data presented here imply that screening programmes should allow evaluation of the entire colon rather than the distal 25 cm. PMID- 15352747 TI - Paravertebral anaesthesia for breast surgery an initial experience at the University Hospital of the West Indies. AB - Paravertebral blockade (PVB) is a regional anaesthetic technique that allows the injection of local anaesthetic agents into the paravertebral space. It has been used for acute and chronic pain relief and as an anaesthetic technique for unilateral surgery of the chest, breast, shoulder, kidney, and inguinal region. Paravertebral blockade has been performed on a limited basis for breast surgery at the University Hospital of the West Indies (UHWI) since 1998. This retrospective review was undertaken to report the initial experience with this block. We reviewed the notes of all patients who were given a PVB alone, or in combination with general anaesthesia (GA). Twenty-one patients had P VB: twenty females and one male, with age range of 24 to 90 years. Six were attempted with PVB alone, but two of these needed supplementation with a GA. Fifteen were done in combination with GA. No complications were recorded The initial experience shows that the performance of PVB is both possible and safe; it may offer an alternative to GA for breast surgery. A randomized prospective study is underway to allow a detailed comparison between the two methods. PMID- 15352748 TI - Thirty-five years of cardiac surgery in Jamaica. AB - A retrospective review of the files of all patients who underwent cardiac surgery at the University Hospital of the West Indies (UHWI) and the Bustamante Hospital for Children (BHC), during the period April 1968 to June 2003 was undertaken. Data collected included age, gender New York Heart Association risk score, type and date of cardiac surgery. The mortality rate of patients who underwent surgery during the period January 1994 to June 2003 was also analyzed A total of 2202 patients had undergone cardiac surgery (CS) in Jamaica during the study period of 35 years and two months. The common surgical procedures were valve surgery- replacement and repair (37.65), correction of patent ductus arteriosus (25.2%) and repair of congenital heart disease (24.2%). Coronary arterial bypass grafting procedures constituted a small percentage (4.1%) of the cardiac surgical operations. A considerable number of patients have undergone CS in Jamaica, but much more needs to be done as the patient load exists. The future of the cardiac surgical service therefore depends on improvement in the facilities at both institutions and the cadre of the intensive care nursing staff The building of the Cardiothoracic-Neurosurgical Unit (commenced in March, 2003) is an essential step towards this. PMID- 15352749 TI - A history of cardiology in Jamaica. AB - The history of cardiology in Jamaica is conveniently considered in decades beginning in the 1950s. The decade of the 1950s was characterized by early descriptions of the pattern of cardiac disease in adults and children in Jamaica, the establishment of a cardiac clinic at the University Hospital of the West Indies and early cardiac surgical landmarks. Extensive preparatory experimental work in the canine laboratory with respect to cardiopulmonary bypass in the early to mid-1960s culminated in the successful completion of the first open heart surgical procedure in April, 1968. Cardiac catheterization was also increasingly developed in the decade of the 1960s. A highlight of the decade of the 1970s was the establishment of the Heart Foundation of Jamaica which began contributing greatly to preventive cardiology in Jamaica by providing a variety of programmes of prevention. In the decade of the 1980s, non-invasive cardiac diagnostic facilities in Jamaica were considerably enhanced by the introduction and development of echocardiography, treadmill exercise testing and ambulatory electrocardiography. In addition, the very important National Rheumatic Fever prevention programme was established. The cardiac catheterization laboratory was re-opened in the 1990s, thus allowing the performance of coronary arteriography in Jamaica for the first time, and interventional cardiology procedures soon followed. The Jamaica Foundation for Cardiac disease was also established in this decade. The vision for the new millennium of "A heart healthy Jamaica in the 21st century" is achievable, but will require appropriate emphasis on expanded preventive and curative cardiology programmes. PMID- 15352750 TI - Dyspepsia: review of management options. AB - Dyspepsia is a common clinical problem which is associated with significant healthcare cost. The management of patients presenting with dyspepsia remains controversial. It has been shown to be cost effective and associated with long term improvement to test and treat young patients positive for H. pylori, who present to the primary care setting. Patients negative for H. pylori may be treated with an antisecretory medication. Prompt endoscopy is recommended in patients over age 50 years, those with alarm symptoms and those who have failed previous therapy. In the Caribbean, the management of patients should be individualized in view of the relatively high prevalence of H pylori infection and gastric carcinoma. PMID- 15352751 TI - Role of yoga in stress management. AB - The state of the mind and that of the body are intimately related. If the mind is relaxed, the muscles in the body will also be relaxed. Stress produces a state of physical and mental tension. Yoga, developed thousands of years ago, is recognized as a form of mind-body medicine. In yoga, physical postures and breathing exercises improve muscle strength, flexibility, blood circulation and oxygen uptake as well as hormone function. In addition, the relaxation induced by meditation helps to stabilize the autonomic nervous system with a tendency towards parasympathetic dominance. Physiological benefits which follow, help yoga practitioners become more resilient to stressful conditions and reduce a variety of important risk factors for various diseases, especially cardio-respiratory diseases. PMID- 15352752 TI - Image and diagnosis. Right diaphragm eventration and bronchopneumonia. PMID- 15352753 TI - The early story of coronary artery disease. AB - Angina pectoris was first adequately described and named in 1772. However, coronary artery disease, as the cause, was noted in 1779. The pathology and clinical features of myocardial infarction were only recognized in 1912. The distinction between angina and infarction was clarified in the 1920s. Nitrate therapy remained the sole treatment until modern medical and surgical developments which this review does not cover. PMID- 15352754 TI - Human external ophthalmomyiasis occurring in Barbados. AB - Human infection with the sheep nasal botfly Oestrus ovis occurs sporadically. In most cases, there is a history of a strike in the eye by the adult fly. Human O. ovis has been reported rarely from the Americas. We report the first case of O. ovis infection in the Caribbean region, which occurred in an urban area of Barbados. The patient responded to removal of the larvae from the conjunctiva and symptomatic treatment. PMID- 15352755 TI - The management of isolated splenic abscess. PMID- 15352756 TI - Beginning of the end? North Mississippi Health Services' settlement of uninsured billing issues raises questions about future of hospital lawsuits. AB - The first settlement in the uninsured billing case could be the sign of a full surrender or just a single defection, healthcare legal experts say. North Mississippi Health Services made the agreement before attorney Richard Scruggs, left, and his firm filed legal action against the system. PMID- 15352757 TI - Eyeing for-profits. Uninsured lawsuits filed against HCA, UHS, HMA. PMID- 15352758 TI - Softer service. CMS eases admission rule, but LTACs still not happy. PMID- 15352759 TI - Tenet takes good with bad. Second quarter brings big loss, but cash flow is up. PMID- 15352760 TI - Still united? N.J. docs say lawsuit stays merger; insurer disagrees. PMID- 15352761 TI - Squaring off. Suits, resignations end another doc, hospital dispute. PMID- 15352762 TI - Unusual case. Oral hearings set in N.Y. for rare antitrust case. PMID- 15352763 TI - Case in point. Challenge of hospital merger key to antitrust climate. PMID- 15352765 TI - Afterglow fading. Democrats' hard sell on healthcare policies fails to generate much of a buzz. PMID- 15352764 TI - The right stuff. U.S. needs to invest in the technology that really pays off. PMID- 15352766 TI - The best care money can buy? The U.S. delights in its medical technology, and pays a premium for it, but studies show other countries beat us in outcomes, values. PMID- 15352767 TI - The need for speed. Rapid blood tests have quickened the diagnostic process, and even faster products are on the way. PMID- 15352768 TI - Combination therapy as initial treatment for hypertension. PMID- 15352769 TI - The health economics of hypertension. PMID- 15352771 TI - 'Imi Hale -- the Native Hawaiian cancer awareness, research, and training network: second-year status report. AB - PURPOSE: The purpose of this paper is to describe 'Imi Hale, a program developed and managed by Native Hawaiians to increase cancer awareness and research capacity among Native Hawaiians. This US subgroup of indigenous people of the Hawaiian islands has disproportionately high rates of cancer mortality and low rates of participation in health and research careers. METHODS: As a community based research project, 'Imi Hale spent its first year gathering data from Native Hawaiians about their cancer awareness and research priorities. These findings guide 'Imi Hale's community and scientific advisors, a community-based Institutional Review Board, Na Liko Noelo (budding researchers), and staff in developing and carrying out projects that address these priority areas. Emphasis is placed on transferring skills and resources to Native Hawaiians through training, technical assistance, and mentorship. A biennial survey assesses the extent to which community-based participatory research principles are being followed. PRINCIPAL FINDINGS: By the end of the school year, statewide and island specific awareness plans were produced, and 9 funded awareness projects are supporting the development and dissemination of Hawaiian health education materials. Research accomplishments include the enrollment of 42 Native Hawaiian Na Liko Noelo (budding researchers), 22 of which are involved in 14 funded research projects. The biennial evaluation survey found that 92% of our advisors felt that 'Imi Hale was promoting scientifically rigorous research that was culturally appropriate and respectful of Native Hawaiian beliefs, and 96% felt that 'Imi Hale was following its own principles of community-based participatory research. CONCLUSION: 'Imi Hale's community-based approach to promoting cancer awareness will result in a sustainable infrastructure for reducing the cancer burden on Native Hawaiians. PMID- 15352772 TI - The Asian American Network for Cancer Awareness, Research and Training's role in cancer awareness, research and training. AB - PURPOSE: The purpose of this paper is to describe the content for the Asian American Network for Cancer Awareness Research and Training (AANCART) with respect to Asian American demographic characteristics and their cancer burden, highlights of accomplishments in various AANCART regions, aspirations for AANCART, and an interim assessment of AANCART's activities to date. METHODS: The author compiled literature and other data references to describe the context for Asian American demographic characteristics and their cancer burden. As the AANCART Principal Investigator, he collected data from internal AANCART reports to depict highlights of accomplishments in various AANCART regions and offer evidence that AANCART's first two specific aims have been attained. PRINCIPAL FINDINGS: With respect to our first specific aim, we have built an infrastructure for cancer awareness, research and training operationally at a Network-wide basis through program directors for biostatistics, community, clinical, and research and in our four original AANCART regions: New York, Seattle, San Francisco, and Los Angeles. With respect to our second specific aim, we have established partnerships as exemplified by working collaboratively with New York's Charles B. Wang Community Health Center in securing external funding with them for a tobacco control initiative and nationally with the American Cancer Society. With respect to our third specific aim, we have been fortunate to assist at least eight junior investigators in receiving NCI-funded pilot studies. The most notable change was the transfer of AANCART's national headquarters from Columbus, Ohio to Sacramento, California along with potentially an increased diversification of Asian American ethnic groups as well as an expansion to Hawaii and Houston. CONCLUSION: As of the end of year 2 of AANCART, AANCART's two specific aims have been achieved. We are focusing on our third specific aim. PMID- 15352773 TI - The Asian Tobacco Education, Cancer Awareness and Research's role in tobacco and cancer control efforts in Asian American communities. AB - PURPOSE: This article identifies the urgent needs for tobacco and cancer control in Asian American communities and the barriers that have historically prevented the development and implementation of tobacco and cancer prevention and intervention research programs among Asian Americans residing in Delaware Valley region of Pennsylvania and New Jersey. This article also presents ATECAR, the first long-term federal effort in tobacco and cancer control targeting the multi ethnic Asian American communities in this area. METHODS: ATECAR uses a three pronged approach: the development of community and university infrastructure, the development of training programs for minority and underserved students so that they will pursue cancer research careers, and community-based intervention research projects. PRINCIPAL FINDINGS: ATECAR's pioneering work redefined our understanding of Asian Americans' smoking habits and the important role of smoking, especially in newly immigrated communities. CONCLUSION: The advent of ATECAR marks the building of a viable infrastructure that can ensure sustainability of current and future efforts in tobacco control and use. PMID- 15352774 TI - The Pacific Islander Cancer Control Network's role in cancer awareness, research and training. AB - PURPOSE: The Pacific Islander Cancer Control Network (PICCN) is one of the 18 Special Populations Networks recently established by the National Cancer Institute (NCI) to address the cancer control needs of America's medically under served populations. The PICCN focuses on three Pacific Islander groups: Samoans, Guamanians/Chamorros, and Tongan Americans. The program provides an infrastructure for collaboration between an academic institution, the University of California, Irvine; an NCI designated cancer center, the Chao Family Comprehensive Cancer Center; community-based organizations; and other agencies concerned with the health of Pacific Islanders. METHODS: The PICCN's objectives include improving cancer awareness, enhancing recruitment to clinical trials, increasing the number of cancer control investigators, and encouraging more research among these Pacific Islanders. The activities that increase cancer awareness include assessing existing cancer education materials aimed at Pacific Islanders, working with the NCI's Cancer Information Service to modify existing materials, developing new culturally-sensitive materials, and distributing the materials in a culturally-sensitive manner. PRINCIPAL FINDINGS: The PICCN enhances recruitment of Pacific Islanders to clinical trials by establishing relationships with cancer centers, making them aware of existing cancer center studies, and developing culturally appropriate recruitment materials when necessary. The network plans to increase the number of Pacific Islander investigators and encourage more research among Pacific Islanders by identifying potential investigators, informing them about existing cancer control training opportunities, developing a new training opportunity, and providing mentors to help with the development of pilot projects and RO1 applications. CONCLUSION: Through these efforts, the PICCN is addressing the goal of eliminating health disparities among ethnic and racial minorities in this country. PMID- 15352775 TI - Perinatal substance use among high risk women in Hawaii: patterns and impact on pregnancy outcomes. AB - PURPOSE: This study examines the relationship between perinatal substance use and adverse pregnancy outcomes among a select group of high risk women residing in the state of Hawaii. METHODS: Participants were the 2,750 women who had live births during 1994 and 1995 and were enrolled in Hawaii's Perinatal Support Services (PSS) program. Demographic and behavioral risk factors associated with selection for participation in the PSS program were examined along with patterns of substance use and adverse perinatal outcomes. PRINCIPAL FINDINGS: After adjustment for demographic factors, Hawaiian/part Hawaiian women compared to non Hawaiian women were 1.5 times as likely to drink alcohol (p<0.01) and 1.8 times as likely to smoke (p<0.001). Among these women, the relative risk (RR) for preterm delivery was significantly higher for those who used alcohol (RR=1.7, p=0.027), tobacco (RR=1.6, p=0.027), or illicit drugs (RR=1.8, 0.029). Among the same women, the relative risk for low birth weight was significantly higher for those who smoked (RR=1.6, p=0.010) or used illicit drugs (RR=1.7, p=0.046). CONCLUSION: Hawaiian/part Hawaiian PSS women had a higher prevalence of substance use than non-Hawaiian PSS women, even after adjusting for age, education, and marital status. RELEVANCE TO ASIAN AMERICAN AND PACIFIC ISLANDER POPULATIONS: Special attention must be paid to the reasons why Hawaiian/part Hawaiian pregnant women use tobacco and alcohol at higher rates than women of other ethnic groups enrolled in Hawaii's PSS Program. Furthermore, services to assist pregnant women refraining from using illicit drugs should be supported. PMID- 15352776 TI - Translation challenges of cross-cultural research and program development. AB - PURPOSE: Translation of cross-cultural surveys and programs pose unique challenges. This paper identifies potential pitfalls of current translation guidelines and presents solutions employed for one project. METHODS: We followed current guidelines for translation of survey instruments including forward and back translation, and also utilized qualitative methods to decenter survey questions. In addition, our project adapted translation by committee with a core translation team of bilingual/trilingual investigators and staff members with diverse skills and complementary degrees of proficiency in the languages and dialects of interest. PRINCIPAL FINDINGS: This approach resulted with a mechanism for emic equivalence, identifying and resolving dialectic and sociolinguistic concerns, selecting appropriate written Chinese styles, and providing appropriate linguistic training to interviewers. CONCLUSIONS: An understanding of the history of a people and its ramifications for the language are important components of the translation process. Allocation of appropriate time, resources, and skills are critical to the successful development of valid cross-cultural survey instruments and effective programs. PMID- 15352777 TI - Strategies in making cross-linked enzyme crystals. PMID- 15352778 TI - Role of structural factors in the nonlinear optical properties of phthalocyanines and related compounds. PMID- 15352779 TI - Comparative quantitative structure-activity relationship studies (QSAR) on non benzodiazepine compounds binding to benzodiazepine receptor (BzR). PMID- 15352780 TI - Alkylidenecarbenes, alkylidenecarbenoids, and competing species: which is responsible for vinylic nucleophilic substitution, [1 + 2] cycloadditions, 1,5-CH insertions, and the Fritsch-Buttenberg-Wiechell rearrangement? PMID- 15352781 TI - Polarization measurements in macro- and micro-Raman spectroscopies: molecular orientations in thin films and azo-dye containing polymer systems. PMID- 15352782 TI - Recent advances in the liquid-phase syntheses of inorganic nanoparticles. PMID- 15352783 TI - Mechanism of oxidation reactions catalyzed by cytochrome p450 enzymes. PMID- 15352784 TI - Manganese clusters with relevance to photosystem II. PMID- 15352785 TI - Biological applications of perturbed angular correlations of gamma-ray spectroscopy. PMID- 15352786 TI - Nanostructured oxides in chemistry: characterization and properties. PMID- 15352787 TI - The Baeyer-Villiger reaction: new developments toward greener procedures. PMID- 15352788 TI - Cyclization reactions of dianions in organic synthesis. PMID- 15352789 TI - Recent developments in the application of oxazoline-containing ligands in asymmetric catalysis. PMID- 15352790 TI - Synthesis and reactivity of Ru-, Os-, Rh-, and Ir-halide-sulfoxide complexes. PMID- 15352791 TI - Freezing on heating of liquid solutions. AB - We report a reversible liquid-solid transition upon heating of a simple solution composed of a-cyclodextrine (alpha CD), water, and 4-methylpyridine. These solutions are homogeneous and transparent at ambient temperature and solidify when heated to temperatures between 45 degrees and 75 degrees. Quasielastic and elastic neutron scattering show that molecular motions are slowed down in the solid and that crystalline order is established. The solution "freezes on heating." This process is fully reversible, on cooling the solid melts. A rearrangement of hydrogen bonds is postulated to be responsible for the observed phenomenon. PMID- 15352792 TI - Reaction-path switching induced by spatial-distribution change of reactants: CO oxidation on Pt(111). AB - We studied the mechanism of CO oxidation on O-covered Pt(111) surfaces during CO exposure by means of time-resolved near edge x-ray absorption fine structure spectroscopy. Two distinct reaction processes were found to occur sequentially; isolated O atoms and island-periphery O atoms contribute to each process. Combination of in situ monitoring of the reaction kinetics and Monte Carlo simulations revealed that CO coadsorption plays a role of inducing the dynamic change in spatial distribution of O atoms, which switches over the two reaction paths. PMID- 15352793 TI - Stimulated emission three-pulse photo-echo peakshift: a mixed pump-probe and photon-echo technique for studying excited-state dynamics. AB - A novel four-pulse photon-echo technique for exploring condensed phase dynamics at different parts of the excited-state potential energy surface is presented. In contrast to traditional three-pulse photon-echo signals, the introduction of a fourth pump pulse allows the use of photon-echo techniques to probe excited-state phenomena. Here, a "proof of principle" experiment is presented where the excited state solvent dynamics of the coumarin 153 chromophore dissolved in methanol is explored. The fluctuations of the stimulated emission transition is probed, in contrast to the ground-state absorption transition explored in traditional echo measurements. Distinctly different excited-state dynamics, in contrast to ground state signals, is observed and discussed. PMID- 15352794 TI - Many-body interaction analysis: algorithm development and application to large molecular clusters. AB - A completely automated algorithm for performing many-body interaction energy analysis of clusters (MBAC) [M. J. Elrodt and R. J. Saykally, Chem. Rev. 94, 1975 (1994); S. S. Xantheas, J. Chem. Phys. 104, 8821 (1996)] at restricted Hartree Fock (RHF)/MA Plesset 2nd order perturbation theory (MP2)/density functional theory (DFT) level of theory is reported. Use of superior guess density matrices (DM's) for smaller fragments generated from DM of the parent system and elimination of energetically insignificant higher-body combinations, leads to a more efficient performance (speed-up up to 2) compared to the conventional procedure. MBAC approach has been tested out on several large-sized weakly bound molecular clusters such as (H(2)O)(n), n=8, 12, 16, 20 and hydrated clusters of amides and aldehydes. The MBAC results indicate that the amides interact more strongly with water than aldehydes in these clusters. It also reconfirms minimization of the basis set superposition error for large cluster on using superior quality basis set. In case of larger weakly bound clusters, the contributions higher than four body are found to be repulsive in nature and smaller in magnitude. The reason for this may be attributed to the increased random orientations of the interacting molecules separated from each other by large distances. PMID- 15352795 TI - Electronic g-tensors of solvated molecules using the polarizable continuum model. AB - We present the implementation of density functional response theory combined with the polarizable continuum model (PCM), enabling first principles calculations of molecular g-tensors of solvated molecules. The calculated g-tensor shifts are compared with experimental g-tensor shifts obtained from electron paramagnetic resonance spectra for a few solvated species. The results indicate qualitative agreement between the calculations and the experimental data for aprotic solvents, whereas PCM fails to reproduce the electronic g-tensor behavior for protic solvents. This failure of PCM for protic solvents can be resolved by including into the model those solvent molecules which are involved in hydrogen bonding with the solute. The results for the protic solvents show that the explicit inclusion of the solvent molecules of the first solvation sphere is not sufficient in order to reproduce the behavior of the electronic g-tensor in protic solvents, and that better agreement with experimental data can be obtained by including the long-range electrostatic effects accounted for by the PCM approach on top of the explicit hydrogen-bonded complexes. PMID- 15352796 TI - Effect of time-dependent basis functions and their superposition error on atom centered density matrix propagation (ADMP): connections to wavelet theory of multiresolution analysis. AB - We present a rigorous analysis of the primitive Gaussian basis sets used in the electronic structure theory. This leads to fundamental connections between Gaussian basis functions and the wavelet theory of multiresolution analysis. We also obtain a general description of basis set superposition error which holds for all localized, orthogonal or nonorthogonal, basis functions. The standard counterpoise correction of quantum chemistry is seen to arise as a special case of this treatment. Computational study of the weakly bound water dimer illustrates that basis set superposition error is much less for basis functions beyond the 6-31+G(*) level of Gaussians when structure, energetics, frequencies, and radial distribution functions are to be calculated. This result will be invaluable in the use of atom-centered Gaussian functions for ab initio molecular dynamics studies using Born-Oppenheimer and atom-centered density matrix propagation. PMID- 15352797 TI - Simplified diagrammatic expansion for effective operators. AB - For a quantum many-body problem, effective Hamiltonians that give exact eigenvalues in reduced model space usually have different expressions, diagrams, and evaluation rules from effective transition operators that give exact transition matrix elements between effective eigenvectors in reduced model space. By modifying these diagrams slightly and considering the linked diagrams for all the terms of the same order, we find that the evaluation rules can be made the same for both effective Hamiltonian and effective transition operator diagrams, and in many cases it is possible to combine many diagrams into one modified diagram. We give the rules to evaluate these modified diagrams and show their validity. PMID- 15352798 TI - Conservation of connectivity of model-space effective interactions under a class of similarity transformation. AB - Effective interaction operators usually act on a restricted model space and give the same energies (for Hamiltonian) and matrix elements (for transition operators, etc.) as those of the original operators between the corresponding true eigenstates. Various types of effective operators are possible. Those well defined effective operators have been shown to be related to each other by similarity transformation. Some of the effective operators have been shown to have connected-diagram expansions. It is shown in this paper that under a class of very general similarity transformations, the connectivity is conserved. The similarity transformation between Hermitian and non-Hermitian Rayleigh Schrodinger perturbative effective operators is one of such transformations and hence the connectivity can be deducted from each other. PMID- 15352799 TI - Thermodynamics of attractive hard rods: a test of mean field density functional theory. AB - Mean field density functional theory (MFDFT) has been employed to calculate the free energy of a pair of attractive hard rods on a ring. The results for homogeneous and optimal inhomogeneous density profiles have been compared with the exact free energy as a test of the approach. We discuss the problems in applying MFDFT to small systems and suggest modifications which allow a reasonably accurate treatment of this particular, rather extreme, case. PMID- 15352800 TI - Interpolating moving least-squares methods for fitting potential energy surfaces: applications to classical dynamics calculations. AB - As a continuation of our efforts to develop efficient and accurate interpolating moving least-squares (IMLS) methods for generating potential energy surfaces, we carry out classical trajectories and compute kinetics properties on higher degree IMLS surfaces. In this study, we have investigated the choice of coordinate system, the range of points (i.e., the cutoff radius) used in fitting, and strategies for selections of data points and basis elements. We illustrate and test the method by applying it to hydrogen peroxide (HOOH). In particular, reaction rates for the O-O bond breaking in HOOH are calculated on fitted surfaces using the classical trajectory approach to test the accuracy of the IMLS method for providing potentials for dynamics calculations. PMID- 15352801 TI - New hybrid method for reactive systems from integrating molecular orbital or molecular mechanics methods with analytical potential energy surfaces. AB - A computational approach to calculating potential energy surfaces for reactive systems is presented and tested. This hybrid approach is based on integrated methods where calculations for a small model system are performed by using analytical potential energy surfaces, and for the real system by using molecular orbital or molecular mechanics methods. The method is tested on a hydrogen abstraction reaction by using the variational transition-state theory with multidimensional tunneling corrections. The agreement between the calculated and experimental information depends on the quality of the method chosen for the real system. When the real system is treated by accurate quantum mechanics methods, the rate constants are in excellent agreement with the experimental measurements over a wide temperature range. When the real system is treated by molecular mechanics methods, the results are still good, which is very encouraging since molecular mechanics itself is not at all capable of describing this reactive system. Since no experimental information or additional fits are required to apply this method, it can be used to improve the accuracy of molecular orbital methods or to extend the molecular mechanics method to treat any reactive system with the single constraint of the availability of an analytical potential energy surface that describes the model system. PMID- 15352802 TI - A novel method for calculating rate constants of diffusion-influenced reactions. AB - In the present work we suggest a method for calculating rate constants of chemical processes affected by mobility of reactants. The method is based on the encounter theory. Unlike the widely accepted model of collision complexes it provides a general formal solution for practically arbitrary reaction scheme. PMID- 15352803 TI - The integral encounter theory of multistage reactions containing association dissociation reaction stages. III. Taking account of quantum states of reactants. AB - The formalism developed in Part I [K. L. Ivanov, N. N. Lukzen, A. A. Kipriyanov, and A. B. Doktorov, Phys. Chem. Chem. Phys. 6, 1706 (2004)] of the present contribution is extended to treat the reacting particles with internal quantum states. Initial spatial correlations of reactants are considered in the framework of this formalism as well. PMID- 15352804 TI - Inelastic hard rods in a periodic potential. AB - A simple model of inelastic hard rods subject to a one-dimensional array of identical wells is introduced. The energy loss due to inelastic collisions is balanced by the work supplied by an external stochastic heat bath. We explore the effect of the spatial nonuniformity on the steady states of the system. The spatial variations of the density, granular temperature, and pressure induced by the gradient of the external potential are investigated and compared with the analogous variations in an elastic system. Finally, we study the clustering process by considering the relaxation of the system starting from a uniform homogeneous state. PMID- 15352805 TI - Solvent effects on electronic properties from Wannier functions in a dimethyl sulfoxide/water mixture. AB - We present an efficient implementation for the calculation of maximally localized Wannier functions (MLWFs) during parallel Car-Parrinello molecular dynamics simulations. The implementation is based on a block Jacobi method. The calculation of MLWFs results in only a moderate (10%-20%) increase in computer time. Consequently it is possible to calculate MLWFs routinely during Car Parrinello simulations. The Wannier functions are then applied to derive molecular dipole moments of dimethyl sulfoxide (DMSO) in gas phase and aqueous solution. We observe a large increase of the local dipole moment from 3.97 to 7.39 D. This large solvent effect is caused by strong hydrogen bonding at the DMSO oxygen atom and methyl groups. Decomposing the dipole moment into local contributions from the S-O bond and the methyl groups is used to understand the electrostatic response of DMSO in aqueous solution. A scheme is given to derive charges on individual atoms from the MLWFs using the D-RESP methodology. The charges also display large solvent effects and give insight into the transferability of recent force field models for DMSO. PMID- 15352806 TI - Calculating expectations with time-dependent perturbations in quantum Monte Carlo. AB - We show that a small perturbation periodic in imaginary time can be used to compute expectation values of nondifferential operators that do not commute with the Hamiltonian within the framework of quantum diffusion Monte Carlo. Some results for the harmonic oscillator and the helium atom are presented showing the validity of the proposed method. PMID- 15352807 TI - Accurate vibrational-rotational partition functions and standard-state free energy values for H2O2 from Monte Carlo path-integral calculations. AB - Accurate quantum mechanical partition functions and absolute free energies of H(2)O(2) are determined using a realistic potential energy surface [J. Koput, S. Carter, and N. C. Handy, J. Phys. Chem. A 102, 6325 (1998)] for temperatures ranging from 300 to 2,400 K by using Monte Carlo path integral calculations with new, efficient polyatomic importance sampling methods. The path centroids are sampled in Jacobi coordinates via a set of independent ziggurat schemes. The calculations employed enhanced-same-path extrapolation of trapezoidal Trotter Fourier path integrals, and the paths were constructed using fast Fourier sine transforms. Importance sampling was also used in Fourier coefficient space, and adaptively optimized stratified sampling was used in configuration space. The free energy values obtained from the path-integral calculations are compared to separable-mode approximations, to the Pitzer-Gwinn approximation, and to values in thermodynamic tables. Our calculations support the recently proposed revisions to the JANAF tables. PMID- 15352808 TI - Analytical second-order geometrical derivatives of energy for the self-consistent charge density-functional tight-binding method. AB - Analytical formulation of the second-order geometrical derivatives of energy for the self-consistent-charge density-functional tight-binding (SCC-DFTB) method is presented. To test its quality and numerical performance, the derived formalism has been coded and applied for calculation of harmonic vibrational frequencies for a set of 17 small and medium size molecules. For this set, the average absolute deviation from experiment is 99 cm(-1) for SCC-DFTB vs 62 cm(-1) for the Moller-Plesset second-order perturbation theory with the cc-pVDZ basis set (MP2/cc-pVDZ) and 32 cm(-1) for the B3LYP density functional method with the same basis set (B3LYP/cc-pVDZ), while the maximal deviation is 465 cm(-1) vs 1,741 cm( 1) for MP2/cc-pVDZ and 112 cm(-1) for B3LYP/cc-pVDZ. The SCC-DFTB results are in reasonable agreement with experiments as well as with ab initio and density functional results, and are better than other semiempirical methods. The SCC-DFTB method allows for considerable computational time saving when compared to other methods while retaining similar overall accuracy. Data for a series of conjugated polyenes show that an analytical formulation of SCC-DFTB is noticeably faster than its numerical formulation. PMID- 15352809 TI - Modeling vibrational spectra using the self-consistent charge density-functional tight-binding method. I. Raman spectra. AB - An extension of the self-consistent charge density-functional tight-binding (SCC DFTB) method is presented that allows for calculating intensities of peaks in vibrational Raman spectra for very large molecules. The extension is based on a simple ansatz: an extra term, which describes interaction of an external electric field with induced atomic charges, is added to the SCC-DFTB energy expression. We apply the modified SCC-DFTB formalism for reproducing vibrational Raman spectra of 17 organic molecules. The calculated spectra are compared with experiment and with spectra obtained from density functional theory (DFT) calculations. We find that the SCC-DFTB method is capable of reproducing most of the features of experimental Raman spectra. Limitations and advantages of this approach are analyzed and suggestions for interpreting calculated SCC-DFTB Raman spectra are given. PMID- 15352810 TI - Rovibrational distributions of HF in the photodissociation of vinyl fluoride at 193 nm: a direct MP2 quasiclassical trajectory study. AB - Quasiclassical trajectory calculations were performed to calculate rovibrational distributions of the nascent HF fragment in the photodissociation of vinyl fluoride at 193 nm. The trajectories were initiated at the transition states of the four-center (4C) and three-center (3C) HF elimination channels, using a microcanonical, quasiclassical normal-mode sampling. In general, the calculated distributions are in reasonably good agreement with experiment. In particular, the trajectory distributions show bimodal character, although not as pronounced as that observed experimentally. The calculations predict that the 3C and 4C distributions are rather similar to each other, which suggests that the low-J and high-J components of the rotational distributions cannot be specifically assigned to each of these channels. PMID- 15352811 TI - Role of the F spin-orbit excited state in the F+HD reaction: contributions to the dynamical resonance. AB - We report quantum mechanical calculations of excitation functions (relative reaction cross sections) for the F+HD reaction. We include three potential energy surfaces and an accurate treatment of all couplings (non-adiabatic, spin-orbit, and Coriolis). Comparison with experimental results [Dong, Lee, and Liu, J. Chem. Phys., 113, 3633 (2000)] show excellent agreement for the DF product channel and an improved but not perfect agreement for the HF product channel. In the former case, when weighted by the (16%) fractional population of the spin-orbit excited state (F(*)) in the beam, the overall reactivity of the F(*) is small (approximately 5%). For the HF product channel and with the same (16%) fractional weight, F(*) reactivity makes a contribution of approximately 12% in the high energy tail of the resonance peak. As a result, averaging over the population of F spin-orbit states in the beam changes the shape of the resonance. The greater the fraction of F(*) in the beam, the less pronounced will be the resonance modulation of the reaction excitation function. PMID- 15352812 TI - Resonance spectrum and dissociation dynamics of ozone in the 3B2 electronically excited state: experiment and theory. AB - The rovibrational spectrum assigned to the low-lying (3)B(2) electronic state of ozone is measured with intracavity laser absorption spectroscopy. The experimental results are interpreted by means of quantum dynamical calculations on a global ab initio potential energy surface. The observed spectrum is shown to originate from the vibrational ground state in the local minimum of the (3)B(2) potential. The spectrum of short-lived resonance states in this local minimum is analyzed. Additionally, the global minimum of the surface is shown to lie in the dissociation channel in the van der Waals region. This region supports a short sequence of weakly bound vibrational states. PMID- 15352813 TI - Dielectric and vibrational properties of amino acids. AB - We calculate polarizability tensors and normal mode frequencies for the amino acids alanine, leucine, isoleucine, and valine using density functional perturbation theory implemented within the plane wave pseudopotential framework. It is found that the behavior of the electron density under external fields depends to a large extent on the geometrical structure of the molecule in question, rather than simply on the constituent functional groups. The normal modes are able to help distinguish between the different types of intramolecular hydrogen bonding present, and help to explain why leucine is found in the zwitterionic form for the gaseous phase. Calculated IR spectra show a marked difference between those obtained for zwitterionic and nonzwitterionic molecules. These differences can be attributed to the different chemical and hydrogen bonds present. Effective dynamical charges are calculated, and compared to atomic charges obtained from Mulliken population analysis. It is found that disagreement exists, largely due to the differing origins of these quantities. PMID- 15352814 TI - Symmetries, vibrational instabilities, and routes to stable structures of clusters of Al, Sn, and As. AB - We investigate the stability of small clusters using density functional theory to compute the total energy, forces, and vibrational frequencies using linear response. We exhibit an efficient and computationally low-cost route to finding stable structures, by starting with high-symmetry structures and distorting them according to their unstable modes. We illustrate this by application to 4-, 6-, and 13-atom clusters of Al, Sn, and As. This technique also naturally elucidates the origins of stability of the lower symmetry structures, which is variously due to the linear or pseudo Jahn-Teller effect, combined with a lowering of various contributions to the total energy. We show that the situation is more complex than has generally been appreciated. PMID- 15352815 TI - Product multiplet branching in the O(1D) + H2-->OH(2Pi) + H reaction. AB - The statistical model of atom-diatom insertion reactions is combined with coupled states capture theory and used to calculate product multiplet-resolved integral cross sections for the title reaction. This involves an ab initio determination of the four electronic potential energy surfaces that correlate with the products ((1,3)A(') and (1,3)A(")), and an accurate description of the electronic and spin orbit couplings between them. The dependence of the resulting cross sections on the final-state rotational quantum number shows a statistical behavior similar to that observed in earlier studies of the reaction in which only the lowest ((1)A(')) potential was retained. In addition, however, the present calculations provide information on the branching between the OH((2)Pi) multiplet levels. Although the two spin-orbit manifolds are predicted to be equally populated, we find a strong propensity for the formation of the Pi(A(')) Lambda-doublet states. These two predictions confirm the experimental results of Butler, Wiesenfeld, Gericke, Brouard, and their co-workers. The nonstatistical population of the OH Lambda-doublet levels is a consequence of the bond breaking in the intermediate H(2)O complex and is preserved through the multiple curve crossings as the products separate. This exit-channel coupling is correctly described by the present theory. PMID- 15352816 TI - On the vertical excitation energy of cyclopentadiene. AB - The vertical excitation energy for the lowest valence pi-->pi(*) transition of cyclopentadiene is investigated. Using a combination of high-level theoretical methods and spectroscopic simulations, the vertical separation at the ground state geometry is estimated to be 5.43+/-0.05 eV. This value is intermediate between those calculated with coupled-cluster and multireference perturbation theory methods and is about 0.13 eV higher than the observed maximum in the absorption profile. PMID- 15352817 TI - Rovibrational and dynamical properties of the hydrogen bonded complex (CH2)2S-HF: a combined free jet, cell, and neon matrix-Fourier transform infrared study. AB - Fourier transform infrared spectra of the nu(s) (HF stretching) band of the (CH(2))(2)S-HF complex have been recorded at 0.1-0.5 cm(-1) resolution in a cooled cell, in a supersonic jet expansion seeded with argon and in a neon matrix at 4.5 K. The combination of controlled temperature effects over a range of 40 250 K and a sophisticated band contour simulation program allows the separation of homogeneous and inhomogeneous contributions and reveals significant anharmonic couplings between intramolecular and intermolecular vibrational modes similar to our previous work on (CH(2))(2)S-DF. The sign of the coupling constants is consistent with the expected strengthening of the hydrogen bond upon vibrational excitation of HF which also explains the observed small variations of the geometrical parameters in the excited state. The analysis of sum and difference combination bands involving nu(s) provides accurate values of intermolecular harmonic frequencies and anharmonicities and a good estimate of the dissociation energy of the complex. Frequencies and coupling parameters derived from gas phase spectra compare well with results from neon matrix experiments. The effective linewidth provides a lower bound for the predissociation lifetime of 10 ps. The comparison between effective linewidths and vibrational densities of states for (CH(2))(2)S-HF and -DF complexes highlights the important role of intramolecular vibrational redistribution in the vibrational dynamics of medium strength hydrogen bonds. PMID- 15352818 TI - Molecular elimination of Br2 in 248 nm photolysis of bromoform probed by using cavity ring-down absorption spectroscopy. AB - By using cavity ring-down spectroscopy technique, we have observed the channel leading to Br(2) molecular elimination following photodissociation of bromoform at 248 nm. A tunable laser beam, which is crossed perpendicular to the photolysis laser beam in a ring-down cell, is used to probe the Br(2) fragment in the B(3)Pi(ou)(+)-X(1)Sigma(g)(+) transition using the range 515-524 nm. The ring down time lasts 500 ns, so the rotational population of the Br(2) fragment may not be nascent nature, but its vibrational population should be. The vibrational population ratio of Br(2)(upsilon=1)/Br(2)(upsilon=0)=0.8+/-0.2 implies that the fragmented Br(2) is vibrationally hot. The quantum yield of the molecular elimination reaction is 0.23+/-0.05, consistent with the values of 0.26 and 0.16 reported in 234 and 267 nm photolysis of bromoform, respectively, using velocity ion imaging. A plausible photodissociation pathway is proposed, based upon this work and ab initio calculations. The A(1)A(2), B(1)E, and C(1)A(1) singlet states of bromoform are probably excited at 248 nm. These excited states may couple to the high vibrational levels of the ground state X(1)A(1) via internal conversion. This vibrationally excited bromoform readily surpasses a reaction barrier 389.6 kJ/mol prior to decomposition. The transition state structure tends to correlate with vibrationally hot Br(2). Dissociation after internal conversion of the excited states to vibrationally excited ground state should result in a large fraction of the available energy to be partitioned in vibrational states of the fragments. The observed vibrationally hot Br(2) fragment seems to favor the dissociation pathway from high vibrational levels of the ground state. Nevertheless, the other reaction channel leading to a direct impulsive dissociation from the excited states cannot be excluded. PMID- 15352819 TI - Conformational effects on vibronic spectra and excited state dynamics of 3 fluorobenzoic acid dimer. AB - Two conformational isomers of 3-fluorobenzoic acid dimer (3-FBA(2)) have been identified in a supersonic jet expansion by use of laser-induced fluorescence excitation (FE), UV-UV hole-burning, and dispersed fluorescence (DF) spectroscopic methods. In the FE spectrum, the S(1) origins of the two isomeric species appear at a frequency gap of only 24 cm(-1), and the vibronic intensities of the redshifted dimer (dimer I) are about two times weaker than those of dimer II. However, ab initio quantum chemistry calculations at the MP2/6-31G(**) level of theory predict that all the isomeric species of 3-FBA(2) have almost the same binding energy (approximately 17 kcal/mol) in the ground state. Furthermore, unlike benzoic acid dimer, the present system shows intense activity for a low frequency mode in both the FE and DF spectra. With the aid of DFT (B3LYP/6 311G(**)) predicted normal mode frequencies, we have assigned the mode to the in plane gear (cogwheel) vibration of the cyclic hydrogen-bonded frame of the dimer. The Franck-Condon profiles for vibronic excitation of the mode indicate that the distortion of the cyclic hydrogen bond frame as a result of S(1)<--S(0) excitation is larger for dimer I than dimer II. Moreover, the fluorescence lifetime at the S(1) zero-point level of the former is also significantly smaller than the latter. Using the predictions of configuration interaction singles calculations, we have proposed that the spectral and dynamical differences between the two isomeric species observed in this study are manifestations of the different characteristics of their S(1) surfaces. By measuring FE, DF, and hole burning spectra of a mixed dimer between 3-fluobenzoic acid and benzoic acid we have shown that the isomeric features in the homodimer spectra are due to two locally excited rotamers of the 3-fluorobenzoic acid moiety. PMID- 15352820 TI - Electron scattering from perfluorocyclobutane (c-C4F8). AB - We report experimental results for electron scattering from perfluorocyclobutane, c-C(4)F(8), obtained from measurements in our two laboratories. A set of differential, integral, and momentum transfer cross sections is provided for elastic scattering for incident electron energies from 1.5 to 100 eV. Inelastic scattering (vibrational excitation) cross sections have been measured for incident electron energies of 1.5, 2, 5, 6, and 7 eV. In order to investigate the role of intermediate negative ions (resonances) in the scattering process we have also measured an excitation function for elastic scattering and vibrational excitation of the ground electronic state of C(4)F(8) for incident energies between 0.6 and 20 eV. These results are compared with the limited amount of data available in the literature for scattering from this molecule. PMID- 15352821 TI - Spin relaxation in isolated molecules and clusters: the interpretation of Stern Gerlach experiments. AB - Intramolecular spin relaxation may occur in isolated molecules or clusters provided that the density of rovibrational eigenstates is sufficiently high to serve as an energy bath and angular momentum is conserved. In the coupled, zero field limit, total angular momentum (J) is the sum of spin (S) and rotational (N) momenta such that J and M(J) are good angular momentum quantum numbers. In the coupled limit, transitions between Zeeman levels (Delta M(J)++0) cannot occur in the absence of an external torque. However, in the high-field limit, J and M(J) are no longer good quantum numbers, as N and S are decoupled and only their projections on the z axis defined by the external field are invariant. In this case M(N) and M(S) remain as good quantum numbers so that angular momentum conserving transitions can occur subject to the selection rule Delta M(N)=-Delta M(S). Determination of the magnetic moments of isolated molecules and clusters via a thermodynamics-based analysis requires that their magnetizations are measured at sufficiently large fields that spin-rotation effects become negligible and the Zeeman level structure approaches the free-spin case. PMID- 15352822 TI - Electronic excitation and charge transfer processes in collisions between Mg(3(1)S0) atoms and Rb+(1S0) ions in the 0.07-4.00 keV energy range. AB - Inelastic collision processes between neutral Mg atoms and Rb(+) ions, both in their ground states, have been studied by means of a crossed molecular beam technique measuring the decay fluorescence of the excited species formed. Emissions corresponding to Mg(3 (1)P(1)), Mg(3 (3)D(3,2,1)), and Mg(4 (3)S(1)), formed by direct target excitation, Rb(5 (2)P(3/2,1/2)), Rb(6 (2)P(3/2,1/2)) produced by electron capture and also the phosphorescent emission due to decay of Mg(3 (3)P(1)), have been detected and the corresponding absolute cross-section values measured both as total values and resolved into their J states. No polarization measurements could be made. Ab initio calculations using pseudopotentials have been performed and from these a manifold of adiabatic energy curves correlating with the different entry and exit channels have been obtained, allowing to propose a qualitative interpretation of the results, such as the shape of the cross section vs energy for different transitions and the oscillating nature of the branching ratios due to interference effects. PMID- 15352823 TI - Rotational excitations of N2O in small helium clusters and the role of Bose permutation symmetry. AB - We present a detailed study of the energetics, structures, and Bose properties of small clusters of (4)He containing a single nitrous oxide (N(2)O) molecule, from N=1 (4)He up to sizes corresponding to completion of the first solvation shell around N(2)O (N=16 (4)He). Ground state properties are calculated using the importance-sampled rigid-body diffusion Monte Carlo method, rotational excited state calculations are made with the projection operator imaginary time spectral evolution method, and Bose permutation exchange and associated superfluid properties are calculated with the finite temperature path integral method. For N< or =5 the helium atoms are seen to form an equatorial ring around the molecular axis, at N=6 helium density starts to occupy the second (local) minimum of the N(2)O-He interaction at the oxygen side of the molecule, and N=9 is the critical size at which there is onset of helium solvation all along the molecular axis. For N> or =8 six (4)He atoms are distributed in a symmetric, quasirigid ring around N(2)O. Path integral calculations show essentially complete superfluid response to rotation about the molecular axis for N> or =5, and a rise of the perpendicular superfluid response from zero to appreciable values for N> or =8. Rotational excited states are computed for three values of the total angular momentum, J=1-3, and the energy levels fitted to obtain effective spectroscopic constants that show excellent agreement with the experimentally observed N dependence of the effective rotational constant B(eff). The non monotonic behavior of the rotational constant is seen to be due to the onset of long (4)He permutation exchanges and associated perpendicular superfluid response of the clusters for N> or =8. We provide a detailed analysis of the role of the helium solvation structure and superfluid properties in determining the effective rotational constants. PMID- 15352824 TI - Can density functional methods be used for open-shell actinide molecules? Comparison with multiconfigurational spin-orbit studies. AB - The geometries, electronic structures, and vibrational frequencies of two isoelectronic compounds PuO(2)(2+) and PuN(2) have been studied in detail at the density functional theory (DFT) and multiconfigurational ab initio levels of theory. Dynamic correlation was taken into account using second-order perturbation theory (CASPT2) and the variational difference-dedicated configuration interaction method for comparison with the results of the DFT study. Spin-orbit effects were included within the framework of an effective uncontracted spin-orbit configuration-interaction method which considers electron correlation effects and spin-orbit coupling on equal footing. The twelve lowest f f electronic transitions are reported. The electronic ground state of both systems is found to be the Omega=4 component of (3)H(g). We thus disagree with an earlier assignment of the ground state of PuN(2) [E. F. Archibong and A. K. Ray, J. Mol. Struct: THEOCHEM 530, 165 (2000)]. Spin-orbit effects are small on both the geometry and vibrational frequencies of the ground states of PuO(2)(2+) and PuN(2), but they completely change the distribution of electronically excited states. A comparison of results obtained with the two classes of methods allows us to demonstrate that an unambiguous assignment of the electronic ground state and electronic spectra requires the use of multireference methods including spin orbit coupling. Single-reference methods such as DFT provide a reasonable description of the electronic properties of ground states of these open-shell systems, and therefore also of their structural and vibrational properties. The experimental antisymmetric stretching frequency of matrix-isolated PuN(2) is reproduced well by both CASPT2 and DFT calculations; generalized gradient approximation formulations of DFT are more successful than hybrid versions in this respect. Ground-state properties of UO(2) (2+), UN(2), UO(2), PuO(2) (2+), and PuN(2) are compared and discussed. PMID- 15352825 TI - Mechanism of anion formation in C 1s-->pi*-excited carbon dioxide. AB - The mechanism of anion formation from core-excited carbon dioxide is investigated in the C 1s-excitation regime (280-340 eV), where negative-ion-positive-ion coincidences and negative-ion-positive-ion-positive-ion coincidences experiments are performed. O(-) formation occurs efficiently upon C 1s-->pi(*)-excitation (290.7 eV) from the singly charged cation CO(2) (+). This anion is measured in coincidence with the singly charged atoms C(+) and O(+). The formation of atomic cations is accompanied by a substantial kinetic energy release. This suggests that an intermediate CO(++) is formed together with O(-), where the dication decays via fission. The results are discussed in terms of an anion formation mechanism from core-excited molecules considering previous experimental results. PMID- 15352826 TI - Bridge mediated two-electron transfer reactions: on the influence of intersite Coulomb interactions. AB - Donor-acceptor two-electron transfer (TET) mediated by a linear molecular bridge is described theoretically. The particular case is considered where the TET takes place in the presence of a strong electronic intersite coupling within the bridge and against the background of fast vibrational relaxation processes. For such a situation the coarse-grained description of bridge-assisted electron transfer in molecular systems can be utilized [Petrov et al., J. Phys. Chem. B 106, 3092 (2002)]. In the present case it leads to kinetic equations and rate expression for TET reactions. Our recent treatment of completely nonadiabtic TET reactions [Petrov et al., J. Chem. Phys. 120, 4441 (2004)] including a reduction to single exponential kinetics (with overall transfer rate K(TET)) is generalized here to the case of strong intrabridge coupling and the presence of intersite Coulomb interactions. The dependence of K(TET) on the bridge length which is determined by a separate stepwise and concerted contribution is discussed in detail. It is found that the intersite Coulomb interaction favors the TET if the donor and the acceptor are uncharged in their completely reduced states (with two excess electrons present). PMID- 15352827 TI - Comparison of lattice and continuum treatments of n-alkanes with experimental data. AB - Using a homologous series of n-alkanes as a model system, we compare the predictions of a lattice Born-Green-Yvon (BGY) theory and a continuum BGY theory with experimental results. We find that both theories are capable of describing the fluid properties and critical points of alkanes ranging from heptamers (n-C7) to nonadecamers (n-C19). We probe the connection between the lattice and continuum BGY models and extend our discussion to include a sampling of other lattice and continuum treatments. PMID- 15352828 TI - The workings of a molecular thermometer: the vibrational excitation of carbon tetrachloride by a solvent. AB - An intriguing energy-transfer experiment was recently carried out in methanol/carbon tetrachloride solutions. It turned out to be possible to watch vibrational energy accumulating in three of carbon tetrachloride's modes following initial excitation of O-H and C-H stretches in methanol, in effect making those CCl(4) modes "molecular thermometers" reporting on methanol's relaxation. In this paper, we use the example of a CCl(4) molecule dissolved in liquid argon to examine, on a microscopic level, just how this kind of thermal activation occurs in liquid solutions. The fact that even the lowest CCl(4) mode has a relatively high frequency compared to the intermolecular vibrational band of the solvent means that the only solute-solvent dynamics relevant to the vibrational energy transfer will be extraordinarily local, so much so that it is only the force between the instantaneously most prominent Cl and solvent atoms that will significantly contribute to the vibrational friction. We use this observation, within the context of a classical instantaneous-pair Landau-Teller calculation, to show that energy flows into CCl(4) primarily via one component of the nominally degenerate, lowest frequency, E mode and does so fast enough to make CCl(4) an excellent choice for monitoring methanol relaxation. Remarkably, within this theory, the different symmetries and appearances of the different CCl(4) modes have little bearing on how well they take up energy from their surroundings--it is only how high their vibrational frequencies are relative to the solvent intermolecular vibrational band edge that substantially favors one mode over another. PMID- 15352829 TI - Dissipative curve crossing problem. I. High-barrier crossing. AB - Variational approach based on the diabatic variational transition state theory optimization is extended and applied to the calculation of the radiationless transition rate for asymmetric high-barrier normal and inverted crossings. Scaling argument is employed to derive an analytic expression for the renormalized frequency as a function of the asymmetry of the crossing and of the dissipation strength for the particular case of Ohmic dissipation (Debye relaxation). The effect of the asymmetry of the crossing on the physical parameters and on the radiationless transition rate is explored. The effective adiabaticity parameter increases with the increasing asymmetry of the crossing in the normal region and decreases with the increasing asymmetry in the inverted region. The physical behavior of the radiationless transition rate in the normal and inverted regions is qualitatively different in the strong dissipation (Smoluchowski) limit. In the inverted region the rate exhibits stretched exponential decay as a function of the dissipation strength reflecting adiabatic suppression in the strong dissipation limit. The accuracy of the approach decreases with the increasing (decreasing) asymmetry of the crossing in the normal (inverted) region and breaks down in the activationless case. PMID- 15352830 TI - An improved renormalization group theory for real fluids. AB - On the basis of White's theory, an improved renormalization group (RG) theory is developed for chain bonding fluids inside the critical region. Outside the critical region, the statistical associating fluid theory based on the first order mean sphere approximation [Fluid Phase Equilibria 171, 27 (2000)] is adopted and all the microscopic parameters are taken directly from its earlier application of real fluids. Inside the critical region, the RG transformation for long-range density fluctuation is derived in the k space, which illustrates explicitly the contributions from the mean-field term, the local density fluctuation, and the nonlocal density fluctuation. The RG theory is applied to describe physical behavior of ten n alkanes (C1-C10) both near to and far from the critical point. With no additional parameters for chain bonding fluids, good results are obtained for critical specific heat and phase coexistence curves and the resulting critical exponents are in good agreement with the reported nonclassic values. PMID- 15352831 TI - Strong enhancement of vibrational relaxation by Watson-Crick base pairing. AB - We have studied the ultrafast dynamics of NH-stretch vibrational excitations in Watson-Crick base pairs consisting of adenine and uracil derivatives. To estimate the influence of the A:U hydrogen bonding on the vibrational dynamics, we have also studied the uracil derivative in monomeric form. The vibrational relaxation of the NH-stretching mode is found to occur much faster in the Watson-Crick base pair than in monomeric uracil. From the delay dependence of the transient vibrational spectra, it can be concluded that both in base-paired and monomeric uracil, the energy relaxation takes place in two steps, the first step being a rapid transfer of energy from the NH-stretching mode to an accepting mode, the second step the relaxation of this accepting mode. The transient spectra show evidence that in the base pair the hydrogen bond between the nucleobases acts as the accepting mode, and that the hydrogen bonding between the bases is responsible for the extremely fast vibrational relaxation in this system. PMID- 15352832 TI - Electron spin relaxation due to reorientation of a permanent zero field splitting tensor. AB - Electron spin relaxation of transition metal ions with spin S> or =1 results primarily from thermal modulation of the zero field splitting (zfs) tensor. This occurs both by distortion of the zfs tensor due to intermolecular collisions and, for complexes with less than cubic symmetry, by reorientational modulation of the permanent zfs tensor. The reorientational mechanism is much less well characterized in previous work than the distortional mechanism although it is an important determinant of nuclear magnetic resonance (NMR) paramagnetic relaxation enhancement phenomena (i.e., the enhancement of NMR relaxation rates produced by paramagnetic ions in solution or NMR-PRE). The classical density matrix theory of spin relaxation does not provide an appropriate description of the reorientational mechanism at low Zeeman field strengths because the zero-order spin wave functions are stochastic functions of time. Using spin dynamics simulation techniques, the time correlation functions of the spin operators have been computed and used to determine decay times for the reorientational relaxation mechanism for S=1. In the zfs limit of laboratory field strengths (H(Zeem)< or =H(Zeem), and that its neglect can lead to serious errors in the interpretation of NMR-PRE data. PMID- 15352833 TI - Mobility-induced instability and pattern formation in a reaction-diffusion system. AB - Ions undergoing a reaction-diffusion process are susceptible to electric field. We show that a constant external field may induce a kind of instability on the state stabilized by diffusion in a reaction-diffusion system giving rise to formation of pattern even when the diffusion coefficients of the reactants are equal. The origin of the pattern is due to the difference in mobilities of the two species and is thus markedly different from that of deformed Turing pattern in presence of the field. While this differential flow instability had been shown earlier to result in traveling waves, we realize in the context of stationary pattern formation in a typical reaction-diffusion-advective system. Our analysis is based on a numerical simulation of a generic model on a two-dimensional domain. PMID- 15352834 TI - Towards an assessment of the accuracy of density functional theory for first principles simulations of water. II. AB - A series of 20 ps ab initio molecular dynamics simulations of water at ambient density and temperatures ranging from 300 to 450 K are presented. Car-Parrinello (CP) and Born-Oppenheimer (BO) molecular dynamics techniques are compared for systems containing 54 and 64 water molecules. At 300 K, an excellent agreement is found between radial distribution functions (RDFs) obtained with BO and CP dynamics, provided an appropriately small value of the fictitious mass parameter is used in the CP simulation. However, we find that the diffusion coefficients computed from CP dynamics are approximately two times larger than those obtained with BO simulations for T>400 K, where statistically meaningful comparisons can be made. Overall, both BO and CP dynamics at 300 K yield overstructured RDFs and slow diffusion as compared to experiment. In order to understand these discrepancies, the effect of proton quantum motion is investigated with the use of empirical interaction potentials. We find that proton quantum effects may have a larger impact than previously thought on structure and diffusion of the liquid. PMID- 15352835 TI - Monte Carlo simulations of segregation in Pt-Re catalyst nanoparticles. AB - We have investigated the segregation of Pt atoms to the surfaces of Pt-Re nanoparticles using the Monte Carlo method and modified embedded-atom method potentials that we have developed for Pt-Re alloys. The Pt(75)Re(25) nanoparticles (containing from 586 to 4,033 atoms) are assumed to have disordered fcc configurations and cubo-octahedral shapes (terminated by [111] and [100] facets), while the Pt(50)Re(50) and Pt(25)Re(75) nanoparticles (containing from 587 to 4,061 atoms) are assumed to have disordered hcp configurations and truncated hexagonal bipyramidal shapes (terminated by [0001] and [1011] facets). We predict that due to the segregation process the equilibrium Pt-Re nanoparticles would achieve a core-shell structure, with a Pt-enriched shell surrounding a Pt-deficient core. For fcc cubo-octahedral Pt(75)Re(25) nanoparticles, the shells consist of almost 100 at. % of Pt atoms. Even in the shells of hcp truncated hexagonal bipyramidal Pt(50)Re(50) nanoparticles, the concentrations of Pt atoms exceed 85 at. % (35 at. % higher than the overall concentration of Pt atoms in these nanoparticles). Most prominently, all Pt atoms will segregate to the surfaces in the hcp truncated hexagonal bipyramidal Pt(25)Re(75) nanoparticles containing less than 1000 atoms. We also find that the Pt atoms segregate preferentially to the vertex sites, less to edge sites, and least to facet sites on the shell of Pt-Re nanoparticles. PMID- 15352836 TI - Gravity-induced liquid crystal phase transitions of colloidal platelets. AB - The influence of gravity on a suspension of sterically stabilized colloidal gibbsite platelets is studied. An initially isotropic-nematic biphasic sample of such a suspension develops a columnar phase on the bottom on prolonged standing. This phenomenon is described using a simple osmotic compression model. We performed Monte Carlo simulations of cut spheres with aspect ratio L/D=1/15 and took data from the literature to supply the equations of state required for the model. We find that the model describes the observed three-phase equilibrium quite well. PMID- 15352837 TI - Electric field induced switching of poly(ethylene glycol) terminated self assembled monolayers: a parallel molecular dynamics simulation. AB - Effects of electric field on the structure of poly(ethylene glycol) (PEG) terminated alkanethiol self-assembled monolayer (SAM) on gold have been studied using parallel molecular dynamics method. An applied electric field triggers a conformational transition from all-trans to a mostly gauche conformation. The polarity of the electric field has a significant effect on the surface structure of PEG leading to a profound effect on the hydrophilicity of the surface. The electric field applied antiparallel to the surface normal causes a reversible transition to an ordered state in which the oxygen atoms are exposed. On the other hand, an electric field applied in a direction parallel to the surface normal introduces considerable disorder in the system and the oxygen atoms are buried inside. The parallel field affects the overall tilt structure of SAMs more adversely than the antiparallel field. PMID- 15352838 TI - The behavior of mixed-metal oxides: physical and chemical properties of bulk Ce1 xTbxO2 and nanoparticles of Ce1-xTbxOy. AB - The physical and chemical properties of bulk Ce(1-x)Tb(x)O(2) and Ce(1 x)Tb(x)O(y) nanoparticles (xTb exchange nor the introduction of oxygen vacancies in Ce(1-x)Tb(x)O(y) significantly affect the charge on the Ce cations. In contrast, the O K-edge and Tb L(III)-edge XANES spectra for Ce(1-x)Tb(x)O(y) nanoparticles show substantial changes with respect to the corresponding spectra of Ce and Tb single oxide references. The Ce(0.5)Tb(0.5)O(y) compounds exhibit a much larger Tb(3+)/Tb(4+) ratio than TbO(1.7). A comparison with the properties of Ce(1-x)Zr(x)O(y) and Ce(1-x)Ca(x)O(y) shows important differences in the charge distribution, the magnitude of the dopant induced strain in the oxide lattice, and a superior behavior in the case of the Ce(1-x)Tb(x)O(y) systems. The Tb-containing oxides combine stability at high temperature against phase segregation and a reasonable concentration of O vacancies, making them attractive for chemical and catalytic applications. PMID- 15352839 TI - Cycloaddition reactions of cyanogen (C2N2) on the Si(100)-2x1 surface. AB - Multireference as well as density functional theories in combination with the surface integrated molecular orbital molecular mechanics were adopted to study the surface reactions of cyanogens on Si(100)-2x1 surface. Three different products were identified as minima in the initial surface reaction. Among these, the [2+2] product is both kinetically easily accessible and thermodynamically the most stable. Therefore, it can be considered as the experimentally found strongly bound surface species. Unlike other conjugated systems, the [4+2] product is less stable than the [2+2] product. Subsequent surface isomerization studies revealed that kinetically favorable channels exist between the initially formed low temperature species and the high-temperature species, indicating that surface morphology changes gradually as a function of surface temperature. Theses two channels eventually lead to the same final surface products, which is consistent with experiment. Current study shows that the subsequent surface isomerizations are the key reactions to better understand the complex surface structures and their properties. PMID- 15352840 TI - Positronium lifetime in polymers. AB - A model describing the relationship between the orthopositronium lifetime and the volume of a void, located in a synthetic zeolite, is analyzed. Our idea, which allows us to take into account the effects of temperature, comprises the introduction of a non-Hermitian term in the Hamiltonian, which accounts for the annihilation of the orthopositronium. The predictions of the present model are also confronted against an already known experimental result. PMID- 15352841 TI - Low-temperature susceptibility of concentrated magnetic fluids. AB - The initial susceptibility of concentrated magnetic fluids (ferrocolloids) has been experimentally investigated at low temperatures. The results obtained indicate that the interparticle dipole-dipole interactions can increase the susceptibility by several times as compared to the Langevin value. It is shown that good agreement between recent theoretical models and experimental observations can be achieved by introducing a correction for coefficients in the series expansion of susceptibility in powers of density and aggregation parameter. A modified equation for equilibrium susceptibility is offered to sum over corrections made by Kalikmanov (Statistical Physics of Fluids, Springer Verlag, Berlin, 2001) and by B. Huke and M. Lucke (Phys. Rev. E 67, 051403, 2003). The equation gives good quantitative agreement with the experimental data in the wide range of temperature and magnetic particles concentration. It has been found that in some cases the magnetic fluid solidification occurs at temperature several tens of kelvins higher than the crystallization temperature of the carrier liquid. The solidification temperature of magnetic fluids is independent of particle concentration (i.e., magneto-dipole interparticle interactions) and dependent on the surfactant type and carrier liquid. This finding allows us to suggest that molecular interactions and generation of some large-scale structure from colloidal particles in magnetic fluids are responsible for magnetic fluid solidification. If the magnetic fluid contains the particles with the Brownian relaxation mechanism of the magnetic moment, the solidification manifests itself as the peak on the "susceptibility-temperature" curve. This fact proves the dynamic nature of the observed peak: it arises from blocking the Brownian mechanism of the magnetization relaxation. PMID- 15352842 TI - Irreversible versus reversible aggregation: mean field theory and experiments. AB - Colloidal aggregation processes arising at different electrolyte concentrations were studied by means of experiments and confronted with theoretical predictions of different kinetic aggregation models. For this purpose, aqueous dispersions of relatively large polystyrene microspheres were chosen as experimental systems. Aggregation was induced by adding KBr electrolyte to the initially stable particle dispersions. During the aggregation processes, the cluster-size distribution was monitored by means of single cluster light scattering. Analyzing the time evolution of the monomer concentration, we found that the processes arising even at moderate electrolyte concentrations cannot be described by pure time-independent irreversible aggregation models. Hence, alternative models such as time-dependent irreversible aggregation and several reversible aggregation models were also tested. The model that considers a time-dependent sticking probability was found to fit the data quite satisfactorily. Nevertheless, the fitted was so slow that it seems not very likely to find such a behavior in real systems. The aggregation-fragmentation models reported in the literature were unable to reproduce the experimental observations. Hence, a more realistic reversible aggregation model was developed. This model accounts also for reenforced or double bonds between the constituent particles. The corresponding fit improved significantly and reached the same quality as the time-dependent model. Moreover, the obtained fitting parameters were in qualitative agreement with the DLVO predictions and so, reversible aggregation seems to be a more reasonable explanation for the experimental data than time-dependent irreversible aggregation. However, no definite statement on the possible secondary bond fragmentation mechanism may be made since both the applied shear stress in the measuring cell and thermal fluctuations can cause weaker bonds to break. PMID- 15352843 TI - Lanthanide transport in stabilized zirconias: interrelation between ionic radius and diffusion coefficient. AB - The diffusion of all stable lanthanides was measured both in calcia stabilized zirconia (CSZ) and in yttria stabilized zirconia (YSZ) in the temperature range between 1,286 and 1,600 degrees C. The lanthanide diffusion coefficients obtained increase with increasing ionic radius. The experimental activation enthalpy of diffusion is near 6 eV for CSZ and between 4 and 5 eV for YSZ and is not strongly affected by the type of lanthanide. The results were correlated with defect energy calculations of the lanthanide diffusion enthalpy using the Mott-Littleton approach. An association enthalpy of cation vacancies with oxygen vacancies of about 1 eV (96 kJ/mol) was deduced in the case of CSZ, while there is no association in the case of YSZ. Furthermore, the change in diffusion coefficients can be correlated to the interaction parameter for the interaction between the lanthanide oxide with zirconia: The higher the interaction parameter, the higher the lanthanide diffusion coefficient. PMID- 15352844 TI - On the thermodynamic stability of clathrate hydrates IV: double occupancy of cages. AB - We have extended the van der Waals and Platteeuw theory to treat multiple occupancy of a single cage of clathrate hydrates, which has not been taken into account in the original theory but has been experimentally confirmed as a real entity. We propose a simple way to calculate the free energy of multiple cage occupancy and apply it to argon clathrate structure II in which a larger cage can be occupied by two argon atoms. The chemical potential of argon is calculated treating it as an imperfect gas, which is crucial to predict accurate pressure dependence of double occupancy expected at high pressure. It is found that double occupancy dominates over single occupancy when the guest pressure in equilibrium with the clathrate hydrate exceeds 270 MPa. PMID- 15352845 TI - Charge renormalization of charged spheres based on thermodynamic properties. AB - At strong electrostatic coupling, counterions are accumulated in the vicinity of the surface of the charged particle with intrinsic charge Z. In order to explain the behavior of highly charged particles, effective charge Z(*) is therefore invoked in the models based on Debye-Huckel approximation, such as the Derjaguin Landau-Verwey-Overbeek potential. For a salt-free colloidal suspension, we perform Monte Carlo simulations to obtain various thermodynamic properties omega in a spherical Wigner-Seitz cell. The effect of dielectric discontinuity is examined. We show that at the same particle volume fraction, counterions around a highly charged sphere with Z may display the same value of omega as those around a weakly charged sphere with Z(*), i.e., omega(Z)=omega(Z(*)). There exists a maximally attainable value of omega at which Z=Z(*). Defining Z(*) as the effective charge, we find that the effective charge passes through a maximum and declines again due to ion-ion correlation as the number of counterions is increased. The effective charge is even smaller if one adopts the Debye-Huckel expression omega(DH). Our results suggest that charge renormalization can be performed by chemical potential, which may be observed in osmotic pressure measurements. PMID- 15352846 TI - Semiflexible polymers in a random environment. AB - We present using simple scaling arguments and one step replica symmetry breaking a theory for the localization of semiflexible polymers in a quenched random environment. In contrast to completely flexible polymers, localization of semiflexible polymers depends not only on the details of the disorder but also on the ease with which polymers can bend. The interplay of these two effects can lead to the delocalization of a localized polymer with an increase in either the disorder density or the stiffness. Our theory provides a general criterion for the delocalization of polymers with varying degrees of flexibility and allows us to propose a phase diagram for the highly folded (localized) states of semiflexible polymers as a function of the disorder strength and chain rigidity. PMID- 15352847 TI - Competition of hydrophobic and Coulombic interactions between nanosized solutes. AB - The solvation of charged, nanometer-sized spherical solutes in water, and the effective, solvent-induced force between two such solutes are investigated by constant temperature and pressure molecular dynamics simulations of model solutes carrying various charge patterns. The results for neutral solutes agree well with earlier findings, and with predictions of simple macroscopic considerations: substantial hydrophobic attraction may be traced back to strong depletion ("drying") of the solvent between the solutes. This hydrophobic attraction is strongly reduced when the solutes are uniformly charged, and the total force becomes repulsive at sufficiently high charge; there is a significant asymmetry between anionic and cationic solute pairs, the latter experiencing a lesser hydrophobic attraction. The situation becomes more complex when the solutes carry discrete (rather than uniform) charge patterns. Due to antagonistic effects of the resulting hydrophilic and hydrophobic "patches" on the solvent molecules, water is once more significantly depleted around the solutes, and the effective interaction reverts to being mainly attractive, despite the direct electrostatic repulsion between solutes. Examination of a highly coarse-grained configurational probability density shows that the relative orientation of the two solutes is very different in explicit solvent, compared to the prediction of the crude implicit solvent representation. The present study strongly suggests that a realistic modeling of the charge distribution on the surface of globular proteins, as well as the molecular treatment of water, are essential prerequisites for any reliable study of protein aggregation. PMID- 15352848 TI - Using nanoparticles to create self-healing composites. AB - The need for viable materials for optical communications, display technologies, and biomedical engineering is driving the creation of multilayer composites that combine brittle materials, such as glass, with moldable polymers. However, crack formation is a critical problem in composites where thin brittle films lie in contact with deformable polymer layers. Using computer simulations, we show that adding nanoparticles to the polymers yields materials in which the particles become localized at nanoscale cracks and effectively form "patches" to repair the damaged regions. Through micromechanics simulations, we evaluate the properties of these systems in the undamaged, damaged, and healed states and determine optimal conditions for harnessing nanoparticles to act as responsive, self assembled "band aids" for composite materials. The results reveal situations where the mechanical properties of the repaired composites can potentially be restored to 75%-100% of the undamaged material. PMID- 15352850 TI - On the nucleation mechanism of the beta-delta phase transition in the energetic nitramine octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine. PMID- 15352849 TI - Phase behavior of polarizable spherocylinders in external fields. AB - Applied electric fields are known to induce significant changes in the properties of systems of polarizable molecules or particles. For rod-shaped molecules, the field-induced behavior can be rather surprising, as in the case of the negative electric birefringence of concentrated solutions of rodlike polyelectrolytes. We have investigated the interplay of shape anisotropy and field-induced anisotropy in molecular dynamics simulations of systems of polarizable soft spherocylinders in an electric field, in the limit of infinitely anisotropic polarizability, taking full account of mutual induction effects. We find a novel crystalline structure (K(2)) in the high-field limit, whose formation is driven by interactions between induced dipoles. For high pressures, the phase diagram exhibits a polar nematic phase between the hexagonal close-packed crystal phase and the K(2) phase. We also compare this system with an analogous system of spherocylinders with permanent electric dipoles and find that qualitatively similar behavior is obtained in the limit of strong coupling of the permanent dipoles to the external field. PMID- 15352851 TI - A note on geometry relaxation of molecular positron complexes. PMID- 15352853 TI - Rehabilitation of a patient with cleft lip and palate with an extremely edentulous atrophied posterior maxilla using zygomatic implants: case report. AB - OBJECTIVE: This case report describes the clinical and surgical management of a patient with a unilateral alveolar cleft and associated extremely atrophied totally edentulous maxilla. METHOD: Two zygomatic implants and four endosseous oral implants were placed under general anesthesia in a compromised maxilla to rehabilitate a 33-year-old patient with cleft lip and palate. The two specially designed zygomatic implants were utilized to avoid the need for bone grafting in the patient. The final prosthetic rehabilitation was an esthetic and functional maxillary overdenture prosthesis supported by implants. RESULTS: Preliminary results have shown how dental prostheses supported by endosseous implants in grafted alveolar cleft are a reliable possibility in the dental rehabilitation of this malformation. CONCLUSION: The use of zygomatic implants may be considered a reliable alternative to more resource-demanding techniques such as bone grafting in patients with cleft palate. PMID- 15352854 TI - Lacrimoauriculodentodigital syndrome with cleft lip/palate and renal manifestations. AB - OBJECTIVE: To describe the largest family reported with the lacrimoauriculodentodigital (LADD) syndrome. DESIGN: A family study of eight individuals with LADD syndrome. SETTING: Pediatric Clinical Research Center at Oakland Children's Hospital. PATIENTS: Eight individuals in a four-generation family. MAIN OUTCOME MEASURE: A diagnosis of LADD syndrome was determined by the presence of one or more of the characteristic lacrimal duct, auricular, dental, or digital malformations. Tear function and orofacial clefting were also considered in the final diagnosis of LADD syndrome. RESULTS: Affected family members had the characteristic features of LADD syndrome, including cup-shaped ears; lacrimal duct obstruction; and dental, forearm, and digit malformations. In addition to the cardinal features of LADD syndrome, the proband was born with cleft lip and palate. The mother and sister of the proband have hydronephrosis, a rare feature of the disorder. Family members also have features not previously described in LADD syndrome, including vesicoureteral reflux, recurrent urinary tract infections, camptodactyly, distal thumb symphalangism, and a bicornuate uterus. CONCLUSIONS: The findings suggest that the presence of vesicoureteral reflux and recurrent urinary tract infections should be evaluated in LADD syndrome patients and that the distinction of LADD syndrome from ectrodactyly ectodermal dysplasia and clefting syndrome by the absence of clefting may need to be reconsidered. PMID- 15352855 TI - Development and clinical usage of a motion analysis system for the face: preliminary report. AB - OBJECTIVE: To evaluate the motion of the face and jaw of patients with cleft lip and palate, facial palsy, and in patients after reconstruction, a motion analyzing system was developed. The aim of this article was to investigate the accuracy of this system and the possibility of clinical application. METHODS: Markers of 1 to 2 mm were placed on the face, and motion images were obtained by three digital video cameras controlled by a synchronizer and recorded on digital video tape. The image was processed on a personal computer. The markers were automatically tracked across the image sequences, and their three-dimensional coordinates were then calculated. MAIN OUTCOME MEASURES: System accuracy was investigated using a positioning actuator with high accuracy and a known object. In three patients with bilateral cleft lip and palate, lip pursing was analyzed using the aforesaid method. RESULTS AND CONCLUSIONS: The mean differences from the known values to the distances between the tracked sample points and to the mobile distances of the sample points per frame were 0.24 to 0.36 mm and 0.02 to 0.05 mm, respectively. Both results were similar regardless of the mobile speed or direction. In five repeated measurements, the mean differences from the known values as for the distances and the mobile speed ranged from 0.19 to 0.38 mm and from 0.00 to 0.07 mm, respectively. Examination of three patients with bilateral cleft lip and palate indicated the possibility that lip movement could be successfully analyzed using the present system. PMID- 15352856 TI - A comparison of oral-nasal balance patterns in speakers who are categorized as "almost but not quite" and "sometimes but not always". AB - OBJECTIVE: The purpose of this study was to determine whether amplitude or temporal patterns of oral-nasal balance differentiate speakers with cleft palate who are classified as belonging to the "almost but not quite" (ABNQ) and "sometimes but not always" (SBNA) subgroups of marginal velopharyngeal inadequacy. DESIGN: The nasal accelerometric vibrational index (NAVI) was used to measure amplitude and temporal aspects of oral-nasal balance during the productions of oral and nasal syllables, words, and sentences. NAVI measures obtained include mean amplitude, time integral (area under the curve), duration, rise time, and fall time. SETTING: Tertiary care center for patients with cleft palate-craniofacial anomalies. PARTICIPANTS: Seventeen patients with repaired cleft palate who were assigned by perceptual assessment to the ABNQ subgroup and 17 patients who were assigned to the SBNA subgroup. RESULTS: No differences were found between the ABNQ and SBNA subgroups with regard to patterns of nasalization. Further analysis as a function of level of production and phonetic context revealed no differences between the subgroups. CONCLUSIONS: Although clinicians may report perceived differences in the resonance patterns of speakers who fall within the category of marginal velopharyngeal inadequacy, further division into the ABNQ and SBNA subgroups has yet to be validated. PMID- 15352857 TI - Digital three-dimensional photogrammetry: evaluation of anthropometric precision and accuracy using a Genex 3D camera system. AB - OBJECTIVE: To determine the precision and accuracy of facial anthropometric measurements obtained through digital three-dimensional (3D) photogrammetry. DESIGN: Nineteen standard craniofacial measurements were repeatedly obtained on 20 subjects by two independent observers, using calipers and 3D photos (obtained with a Genex 3D camera system), both with and without facial landmarks labeled. Four different precision estimates were then calculated and compared statistically across techniques. In addition, mean measurements from 3D photos were compared statistically with those from direct anthropometry. RESULTS: In terms of measurement precision, the 3D photos were clearly better than direct anthropometry. In almost all cases, the 3D photo with landmarks labeled had the highest overall precision. In addition, labeling landmarks prior to taking measurements improved precision, regardless of method. Good congruence was observed between means derived from the 3D photos and direct anthropometry. Statistically significant differences were noted for seven measurements; however, the magnitude of these differences was often clinically insignificant (< 2 mm). CONCLUSIONS: Digital 3D photogrammetry with the Genex camera system is sufficiently precise and accurate for the anthropometric needs of most medical and craniofacial research designs. PMID- 15352858 TI - Cleft palate in a newborn with duplication 2(q13q23). AB - OBJECTIVES: A preterm boy was born with multiple anomalies including cleft palate and ventricular septal defect. Chromosome analysis on a blood sample identified additional material within the long arm of chromosome 2. SETTING: The newborn was in the neonatal intensive care unit requiring tertiary care during his 22 days of life. RESULTS: A supplementary fluorescent in situ hybridization test was performed to confirm the extra chromosomal material was chromosome 2. Parents' chromosomes were normal, indicating a de novo duplication of 2q13q23. CONCLUSION: Comparison of this case with those in the literature suggests involvement of cleft palate of cases with duplication of 2q13. PMID- 15352859 TI - Abnormal vomer development in human fetuses with isolated cleft palate. AB - OBJECTIVE: The aim of this study was to elucidate the prenatal human development of the vomer with emphasis on the vomeral footplate and to assess vomeral morphology in fetuses with isolated cleft palate. MATERIAL AND METHODS: Nine human fetuses of which four were normal (menstrual age [MA] 13 to 21 weeks) and five with isolated cleft palate (14 to 19 weeks MA) were studied. Midaxial cranial tissue blocks from the fetuses were cut frontally in 4microm serial sections. Sections were stained with toluidine blue in 30% ethanol. RESULTS: From 16 weeks MA, the vomeral footplate of normal fetuses was formed from bilateral ossifications located below a U-shaped vomeral body. Later in development, an osseous connection was found between the footplate and the vomeral body. Neither bilateral areas of ossification below the vomer nor a vomeral footplate was observed in isolated cleft palate fetuses. CONCLUSIONS: In normal fetuses, the base or footplate of the vomeral bone appears from 16 weeks MA in frontal sections. In fetuses with isolated cleft palates, with no connection between the nasal septum and the maxillary processes, this vomeral footplate does not develop in the period observed (14 to 19 weeks MA). PMID- 15352860 TI - Management of the impaired adult cleft patient: the last chance. AB - OBJECTIVE: Unfortunately, not all patients receive the benefits of a well trained, experienced, multidisciplinary cleft team, and a significant number of patients reach adulthood with severe neglected cleft related problems. The vast majority of this group have made their way through several surgical procedures, usually performed by different surgeons in different centers, and still carry the original cleft stigmata aggravated by variable degrees of maxillary hypoplasia, fistulae, scarring, muscle pull, soft tissue tension, lost teeth, dental adaptation, and instability of bony fragments. Adult patients who reach this stage with their original deformities have, in most instances, lost their best opportunity to be properly treated. With this assumption in mind, an aggressive surgical plan focused mainly on the deformities that can objectively achieve functional and aesthetical improvement should be individualized for each patient. In this article, experience with 38 consecutive adult patients with cleft managed following a two-stage protocol used at our cleft clinic for the last 4 years is presented. CONCLUSION: An algorithm to help definitively solve these deformities is suggested. PMID- 15352861 TI - Dentoalveolar growth of patients with complete unilateral cleft lip and palate by early two-stage furlow and push-back method: preliminary results. AB - OBJECTIVE: This study examined dentoalveolar growth changes prior to the time of palatoplasty up to 3 years of age by the early two-stage Furlow and push-back methods. SUBJECTS: Thirty-four Japanese patients with complete unilateral cleft lip and palate (UCLP) treated with either a two-stage Furlow procedure (Furlow group: seven boys, eight girls) from 1998 to 2002 or a push-back procedure (push back group; 12 boys, 7 girls) from 1993 to 1997. METHOD: Consecutive plaster models were measured by three-dimensional laser scanner, before primary palatoplasty, before hard palate closure (Furlow group only), and at 3 years of age. Bite measures were taken at 3 years of age. RESULTS: In the Furlow group, arch length, canine width, first and second deciduous molar width and cross sectional area, and depth and volume at midpoint showed greater growth than in the push-back group. In the Furlow group, the crossbite score was also better than in the push-back group at 3 years of age. In comparison with the push-back group, inhibition of growth impediment in the anterior region was observed in the horizontal direction in the Furlow group. In the midregion, it was observed in the horizontal and vertical directions, and in the posterior region it was observed in the horizontal direction. CONCLUSION: The results demonstrate that the early two-stage Furlow method showed progressive alveolar growth. Therefore, the early two-stage Furlow method is a more beneficial procedure than the push back method. PMID- 15352862 TI - Speech outcome following treatment in cross-linguistic cleft palate studies: methodological implications. AB - OBJECTIVE: In cross-linguistic studies of cleft palate speech outcome following treatment, treatment is the independent variable, speech outcome is the dependent variable, and the speakers' language background is a background variable, like sex, age, and cleft type, which must be eliminated. This article focuses on language as a background variable and how it should be treated. The methodological problems are illustrated through a presentation of two cross linguistic speech outcome studies. CONCLUSION: When speakers of different language background are included in cleft palate studies of speech outcome following treatment, speech outcome data should be based on speech units that are phonetically identical across languages. This affects the make-up of the speech material used in the study. In practice, the requirement of phonetically identical speech units may not be totally met, and detailed information regarding the interaction between the cleft condition and speech sound production is still required to fully understand how the validity of data is affected if this requirement is not met. PMID- 15352863 TI - Prevalence of palatal and alveolar cysts in babies with cleft lip and palate. AB - OBJECTIVE: To evaluate the prevalence of palatal and alveolar cysts in babies with cleft lip and/or palate. DESIGN: Cross-sectional. SETTING: Hospital for Rehabilitation of Craniofacial Anomalies, University of Sao Paulo (HRAC-USP), Bauru, Sao Paulo, Brazil. PARTICIPANTS: Two hundred ninety-one Caucasian babies divided into four groups according to the type of cleft: cleft lip with or without cleft alveolus (70), complete unilateral cleft lip and palate (112), complete bilateral cleft lip and palate (56), and cleft palate (53). RESULTS: A low prevalence of palatal and alveolar cysts was observed among patients with the four different types of clefts, with no statistically significant difference between genders. The maxilla and the anterior area of the mouth were more affected than the mandible and the posterior area. CONCLUSIONS: The low prevalence of palatal and alveolar cysts in the four groups of babies with clefts included in this study may have been due to the high mean age of the sample. PMID- 15352864 TI - An acoustic study of the temporal characteristics of nasalization in children with and without cleft palate. AB - OBJECTIVE: The purpose of this study was to measure the oral and nasal cavity acoustic energies to determine whether temporal patterns of nasalization differentiate children with and without cleft palate and the extent to which vowel context contributes to these temporal differences. DESIGN: Nasal onset interval, nasal offset interval, and total nasalization duration measures were obtained from acoustic waveforms, spectrograms, and energy contours acquired using the Computerized Speech Lab (Kay Elemetrics, Lincoln Park, NJ). In addition to absolute temporal values, proportional durations of nasalization were measured to obtain information regarding the relative duration of nasalization. PARTICIPANTS: Fifteen children with cleft palate exhibiting mild hypernasality and 15 children without cleft palate participated in the study. Children in both groups were between 4 and 7 years of age. RESULTS: Each of the three absolute measures was significantly different between the two groups of subjects and within the three vowel contexts. Children with cleft palate showed longer temporal characteristics than children without cleft palate in all three absolute measures. The three temporal variables of the high vowel contexts were generally longer than those of low vowel contexts. Regarding proportional measures, nasal offset interval ratio, and total nasalization ratio showed significant differences between the two groups. CONCLUSIONS: Nasalization, as reflected by acoustic signals, showed different timing characteristics between children with cleft palate and without cleft palate and across vowel contexts. This suggests that the duration of nasalization reflecting temporal patterns of the oral-nasal acoustic impedance may have an influence on the perception of hypernasality. PMID- 15352865 TI - Variations in the tensor veli palatini muscle with special reference to its origin and insertion. AB - OBJECTIVE: Previous research on the tensor veli palatini muscle (TVP) has produced conflicting descriptions of its functions and topographical relationships with other orofacial structures. The goal of this study was to describe the morphology of the TVP in a systematic and comprehensive manner. METHODS: One hundred nineteen sides of 77 human heads from donated cadavers were partially dissected under a binocular microscope. Histological examination of the hard tissue-muscle interfaces was also undertaken. RESULTS: There were two adjacent origins of the TVP: the cranial base origin (CB origin) and the auditory tube cartilage origin (AT origin). The CB origin always lay anterior to the AT origin and there was no septum or loose tissue between the two muscular laminae leading from these origins. The muscle fibers converged on a central tendinous plate in the muscle belly, which gradually became a common tendon that rounded the pterygoid hamulus before inserting into the palatine aponeurosis. Notably, secondary insertions were found on the maxillary tuber (33.6%) and/or in the submucosal tissue near the palatoglossal arch (37.8%). Maxillary insertions were almost exclusively associated with an AT origin that was wide as or wider than the CB origin. Histological observations confirmed that the hamulus acted purely as a pulley and suggested that a connecting band to the tensor tympani had no or few functions of an intermediate tendon. CONCLUSIONS: The TVP appears to act as the dilator tubae and that this function can be maintained by preserving or reconstructing the maxillary insertion during push-back surgery, even if hamulotomy is necessary. PMID- 15352866 TI - A suspected case of Abruzzo-Erickson syndrome. AB - The features of Abruzzo-Erickson syndrome are described. A case report of an adult female suspected to have this condition is described and illustrated. The possible link to Abruzzo-Erickson syndrome and differential diagnosis of CHARGE is discussed. PMID- 15352867 TI - Kita H, Kochi S, Kamiya N, et al. Alveolar ridge notching as a predictor for secondary bone grafting in incomplete alveolar clefts. PMID- 15352868 TI - Approaches of South African traditional healers regarding the treatment of cleft lip and palate. AB - OBJECTIVE: Due to the large number of Black South Africans known to consult with traditional healers and the influential role they play in the developing world, this study investigated the approaches of a group of 15 traditional healers toward the treatment of cleft lip and palate. DESIGN: An exploratory-descriptive, qualitative research design was employed, which involved the use of standardized, semistructured interviews, conducted with the assistance of an interpreter who was fluent in several African languages. SETTING: Interviews were conducted outdoors in places in which traditional healers usually consult with their patients. These areas were in the fields close to the traditional healers' homes or under a highway bridge in the Johannesburg inner city. PARTICIPANTS: Individual interviews were conducted with 15 African traditional healers who had been practicing in their callings for at least 1 year. RESULTS: Traditional healers interviewed had treated one to six persons with a cleft lip, cleft palate, or both. Most informants believed that clefts were caused by the ancestors, spirits, and witchcraft. A variety of plant and animal products were used to treat these conditions and were augmented by spiritual resources from the ancestors. All except one healer had undergone formal training, although they had received no specific training relative to cleft lip and palate. Most participants reported referring patients to Western health care practitioners who were referred to as modern doctors but did not receive reciprocal referrals from these professionals. Patients generally consulted with traditional healers because this approach was part of their culture. CONCLUSIONS: These findings have important implications for health care professionals as well as traditional healers in terms of cross-cultural consultation, collaboration, and information sharing with regard to cleft lip and palate; the potential use of traditional healers in primary health care and education; and further research. PMID- 15352869 TI - A cephalometric evaluation of lower lip in patients with unilateral cleft lip and palate. AB - OBJECTIVE: The aim of this study was to evaluate cephalometrically the lower lip position and area of patients with unilateral cleft lip and palate (UCLP) comparatively with Class I skeletodental normal subjects. PATIENTS: Lateral cephalometric and hand-wrist radiographs obtained from 24 patients with UCLP (mean age 12.86 years), along with 20 normal individuals (mean age 12.33 years) used as a control group, were examined. DESIGN: In addition to standard cephalometric dentofacial variables, lower lip area (superior, middle, inferior) was also measured using a digital planimeter on the lateral cephalograms. RESULTS: The superior and middle part of the lower lip areas were significantly smaller (p < .05) in the UCLP group, compared to the control group. The inferior and total lower lip areas of patients with UCLP were found to be significantly smaller than controls. The labiomental angle was also smaller (38.79 degrees). CONCLUSIONS: The results suggest that the lower lip of patients with UCLP is smaller, retruded, and curved, with a deep labiomental sulcus, compared with normal individuals during puberty. Therefore, practitioners should focus not only on the upper lip of patients with UCLP but also should consider the lower lip as it was found distinct from normal individuals during puberty. PMID- 15352870 TI - Demographic and reproductive factors associated with hemifacial microsomia. AB - OBJECTIVE: To identify demographic and reproductive risk factors for hemifacial microsomia in offspring. DESIGN: In a case-control study, maternal interviews were conducted within 3 years after delivery. Cases with hemifacial microsomia were ascertained from craniofacial centers in 26 cities in the United States and Canada. Controls were patients of the cases' pediatricians. Two hundred thirty nine cases were compared with 854 controls. Odds ratios for various infant and maternal factors were estimated. RESULTS: Cases had lower birth weights, were more often male or a twin, and had more relatives with craniofacial malformations or hearing loss than controls. Case mothers had lower family incomes, had a lower body mass index, had more vaginal bleeding in the second trimester, and were more likely to have had a spontaneous abortion in a previous pregnancy. CONCLUSIONS: Nonmodifiable factors (age and parity) were not associated with hemifacial microsomia risk. Factors that are related to poverty (low family income, late recognition of pregnancy, and low body mass index) are associated with an increase in risk. High risk estimates for multiple pregnancies and second trimester vaginal bleeding suggest a vascular etiology. PMID- 15352871 TI - Conserved extracellular cysteine residues and cytoplasmic loop-loop interplay are required for functionality of the heptahelical MLO protein. AB - We performed a structure-function analysis of the plasma membrane-localized plant specific barley (Hordeum vulgare) MLO (powdery-mildew-resistance gene o) protein. Invariant cysteine and proline residues, located either in extracellular loops or transmembrane domains that have been conserved in MLO proteins for more than 400 million years, were found to be essential for MLO functionality and/or stability. Similarly to many metazoan G-protein-coupled receptors known to function as homo- and hetero-oligomers, FRET (fluorescence resonance energy transfer) analysis revealed evidence for in planta MLO dimerization/oligomerization. Domain-swap experiments with closely related wheat and rice as well as diverged Arabidopsis MLO isoforms demonstrated that the identity of the C-terminal cytoplasmic tail contributes to MLO activity. Likewise, analysis of a progressive deletion series revealed that integrity of the C-terminus determines both MLO accumulation and functionality. A series of domain swaps of cytoplasmic loops with the wheat (Triticum aestivum) orthologue, TaMLO-B1, provided strong evidence for co operative loop-loop interplay either within the protein or between MLO molecules. Our data indicate extensive intramolecular co-evolution of cytoplasmic domains in the evolutionary history of the MLO protein family. PMID- 15352872 TI - Random mutagenesis of bacterial luciferase: critical role of Glu175 in the control of luminescence decay. AB - Bacterial luciferases (LuxAB) can be readily classed as slow or fast decay luciferases based on their rates of luminescence decay in a single turnover assay. Luciferases from Vibrio harveyi and Xenorhabdus (Photorhabdus) luminescens have slow decay rates, and those from the Photobacterium genus, such as Photobacterium fisheri, P. phosphoreum and P. leiognathi, have rapid decay rates. By substitution of a 67-amino-acid stretch of P. phosphoreum LuxA in the central region of the LuxA subunit, the 'slow' X. luminescens luciferase was converted into a chimaeric luciferase with a significantly more rapid decay rate [Valkova, Szittner and Meighen (1999) Biochemistry 38, 13820-13828]. To understand better the role of specific residues in the classification of luciferases as slow and fast decay, we have conducted random mutagenesis on this region. One of the mutants generated by a single mutation on LuxA at position 175 [E175G (Glu175- >Gly)] resulted in the 'slow decay' X. luminescens luciferase being converted into a luciferase with a significantly more rapid decay rate. These results indicate the importance of Glu175 in LuxA as a critical residue for differentiating between 'slow' and 'fast' luciferases and show that this distinction is primarily due to differences in aldehyde affinity and in the decomposition of the luciferase-flavin-oxygen intermediate. PMID- 15352873 TI - X-ray crystal structure of ornithine acetyltransferase from the clavulanic acid biosynthesis gene cluster. AB - The orf6 gene from the clavulanic acid biosynthesis gene cluster encodes an OAT (ornithine acetyltransferase). Similar to other OATs the enzyme has been shown to catalyse the reversible transfer of an acetyl group from N-acetylornithine to glutamate. OATs are Ntn (N-terminal nucleophile) enzymes, but are distinct from the better-characterized Ntn hydrolase enzymes as they catalyse acetyl transfer rather than a hydrolysis reaction. In the present study, we describe the X-ray crystal structure of the OAT, corresponding to the orf6 gene product, to 2.8 A (1 A=0.1 nm) resolution. The larger domain of the structure consists of an alphabetabetaalpha sandwich as in the structures of Ntn hydrolase enzymes. However, differences in the connectivity reveal that OATs belong to a structural family different from that of other structurally characterized Ntn enzymes, with one exception: unexpectedly, the alphabetabetaalpha sandwich of ORF6 (where ORF stands for open reading frame) displays the same fold as an DmpA (L aminopeptidase D-ala-esterase/amidase from Ochrobactrum anthropi), and so the OATs and DmpA form a new structural subfamily of Ntn enzymes. The structure reveals an alpha2beta2-heterotetrameric oligomerization state in which the intermolecular interface partly defines the active site. Models of the enzyme substrate complexes suggest a probable oxyanion stabilization mechanism as well as providing insight into how the enzyme binds its two differently charged substrates. PMID- 15352874 TI - Dysfunctional proofreading in the Escherichia coli DNA polymerase III core. AB - The epsilon-subunit contains the catalytic site for the 3'-->5' proofreading exonuclease that functions in the DNA pol III (DNA polymerase III) core to edit nucleotides misinserted by the alpha-subunit DNA pol. A novel mutagenesis strategy was used to identify 23 dnaQ alleles that exhibit a mutator phenotype in vivo. Fourteen of the epsilon mutants were purified, and these proteins exhibited 3'-->5' exonuclease activities that ranged from 32% to 155% of the activity exhibited by the wild-type epsilon protein, in contrast with the 2% activity exhibited by purified MutD5 protein. DNA pol III core enzymes constituted with 11 of the 14 epsilon mutants exhibited an increased error rate during in vitro DNA synthesis using a forward mutation assay. Interactions of the purified epsilon mutants with the alpha- and theta;-subunits were examined by gel filtration chromatography and exonuclease stimulation assays, and by measuring polymerase/exonuclease ratios to identify the catalytically active epsilon511 (I170T/V215A) mutant with dysfunctional proofreading in the DNA pol III core. The epsilon511 mutant associated tightly with the alpha-subunit, but the exonuclease activity of epsilon511 was not stimulated in the alpha-epsilon511 complex. Addition of the theta;-subunit to generate the alpha-epsilon511-theta; DNA pol III core partially restored stimulation of the epsilon511 exonuclease, indicating a role for the theta;-subunit in co-ordinating the alpha-epsilon polymerase exonuclease interaction. The alpha-epsilon511-theta; DNA pol III core exhibited a 3.5-fold higher polymerase/exonuclease ratio relative to the wild-type DNA pol III core, further indicating dysfunctional proofreading in the alpha-epsilon511 theta; complex. Thus the epsilon511 mutant has wild-type 3'-->5' exonuclease activity and associates physically with the alpha- and theta;-subunits to generate a proofreading-defective DNA pol III enzyme. PMID- 15352876 TI - Histochemical and morphometrical characterization and distribution of fibre types in four muscles of ostrich (Struthio camelus). AB - A staining procedure used for simultaneously determining three different fibre types in single sections bovine, porcine or ovine skeletal muscle was modified for use with ostrich skeletal muscle. The muscle fibres of gastrocnemius pars externa, tibialis cranialis caput tibiale, tibialis cranialis caput femorale and fibularis longus tendo caudalis were studied. The histochemical results revealed the presence of three types of fibre only in the gastrocnemius pars externa muscle: fast-twitch glycolytic fibres (FG), fast-twitch oxidative glycolytic fibres (FOG) and slow-twitch oxidative fibres (SO), while in the other muscles the FG fibres were absent. The percentage distribution of fibres types showed a higher incidence of SO fibres compared to FOG fibres in tibialis cranialis caput femorale and tibialis cranialis caput tibiale muscles, while it was opposite in the case of the fibularis longus tendo caudalis muscle. In the gastrocnemius pars externa muscle the FG fibres outnumber the other fibres, followed by the SO and FOG fibres. The results of the analysis of variance show significant interaction between muscle x fibre type for every morphometric parameter evaluated. Differences about value of fibres area exists between tibialis cranialis caput femorale and fibularis longus tendo caudalis muscles. Both fibre types in tibialis cranialis caput tibiale muscle have mean values of transversal section area smaller than tibialis cranialis caput femorale. The other morphometric parameters show a similar trend. The gastrocnemius pars externa muscle presents similar dimensions of muscle fibres for the FG and FOG types, and significantly smaller for the SO type. PMID- 15352875 TI - Protein interaction quantified in vivo by spectrally resolved fluorescence resonance energy transfer. AB - We describe a fluorescence resonance energy transfer (FRET)-based method for finding in living cells the fraction of a protein population (alpha(T)) forming complexes, and the average number (n) of those protein molecules in each complex. The method relies both on sensitized acceptor emission and on donor de-quenching (by photobleaching of the acceptor molecules), coupled with full spectral analysis of the differential fluorescence signature, in order to quantify the donor/acceptor energy transfer. The approach and sensitivity limits are well suited for in vivo microscopic investigations. This is demonstrated using a scanning laser confocal microscope to study complex formation of the sterile 2 alpha-factor receptor protein (Ste2p), labelled with green, cyan, and yellow fluorescent proteins (GFP, CFP, and YFP respectively), in budding yeast Saccharomyces cerevisiae. A theoretical model is presented that relates the efficiency of energy transfer in protein populations (the apparent FRET efficiency, E(app)) to the energy transferred in a single donor/acceptor pair (E, the true FRET efficiency). We determined E by using a new method that relies on E(app) measurements for two donor/acceptor pairs, Ste2p-CFP/Ste2p-YFP and Ste2p GFP/Ste2p-YFP. From E(app) and E we determined alpha(T) approximately 1 and n approximately 2 for Ste2 proteins. Since the Ste2p complexes are formed in the absence of the ligand in our experiments, we conclude that the alpha-factor pheromone is not necessary for dimerization. PMID- 15352877 TI - Male and female effects on the in vitro production of bovine embryos. AB - A 3-year study was carried out to evaluate male and female effects on the efficiency of an in vitro fertilization (IVF) programme. The semen of different bulls used for artificial insemination was tested for the in vitro production of transferable blastocysts. The fertilization capacity was recorded for each bull. Bovine oocytes were matured in vitro, fertilized with frozen/thawed semen of 63 individual bulls and cultured during 8 days. The semen of one bull was used as control. The percentage of cleavage (36.3-93.4%) and blastocysts on day 7 (6.9 51.2%) varied from bull to bull. Despite high variability, blastocysts were produced with the semen of all bulls in the first trial. Moreover, oocytes fertilized with 85% of tested bulls reached a blastocyst rate not different to the control bull. The correlation coefficients of six bulls showed no significant male effect but an influence of oocytes on the cleavage rate (F-value 0.38, P > 0.05, and 12.4, P < 0.001, respectively). The development to blastocysts on day 7 was significantly influenced by sperms and also oocytes and session (P < 0.01), but no combined interaction was observed between female and male. It is concluded that transferable embryos can be produced in vitro in the first trial with frozen/thawed semen of 63 tested bulls. The results show different capacities of bulls to produce embryos and high male and female effects on the efficiency of an IVF programme. PMID- 15352878 TI - On the structure of the adrenal gland of the common seal (Phoca vitulina vitulina). AB - The adrenal gland is a vitally important endocrine gland that occupies a central role in the regulatory mechanisms of the body metabolism. Environmental stress factors lead to permanent strain and overload of the body resulting in structural alterations of the adrenals that in turn are followed by hormonal imbalances. This leads to an increased susceptibility to bacterial and viral diseases. The recurrence of numerous fatalities in the different seal populations of the North Sea (during the years 1988, 1989 and 2002), of the Baikal Lake and Caspian Sea (during the years 2000 and 2001) were the motive for a morphological investigation of the species-specific structure of the adrenal gland of the common seal in order to differentiate environmental stress-induced pathological alterations from the physiological structure of this organ. The study was based on adrenals of 112 common seals (Phoca vitulina vitulina) using light microscopic and transmission and scanning electron microscopic methods. The phocine adrenal gland displays several structural characteristics. Originating from the connective tissue organ capsule, narrow and broad septa intersperse the adrenal cortex. These septa contain blastemata as a reserve for the regeneration of hormone-producing cortical cells. Such blastemata are also occurring in the form of an intermediate zone in between the zona glomerulosa and zona fasciculata in the phocine adrenal cortex. Another species-specific characteristic is an inverse part of the adrenal cortex encircling the central vein of the organ. These structural features have to be considered in assessment and definition of pathological alterations of the adrenals as observed in the form of exhausted blastema cell pools in the adrenocortex of seals perished in the mentioned phocine mass mortalities. PMID- 15352879 TI - The neonatal treatment of rats with monosodium glutamate induces morphological changes in the subfornical organ. AB - The parenteral administration of monosodium glutamate (MSG) to neonatal rats induces specific lesions in the central nervous system that lead to a well characterized neuroendocrinological dysfunction. Additionally, it has been shown that MSG-treated rats present a blunted blood pressure response to the injection of nitric oxide synthase inhibitors. Recently, a similar cardiovascular alteration has been reported after the electrolytic lesion of the anteroventral region of the third ventricle affecting the connections of the subfornical organ (SFO). We hypothesized that the treatment of neonatal rats with MSG could affect the nitrergic cells of the SFO. In the present work, we have looked for alterations in the NADPH-diaphorase activity (a commonly used marker for nitrergic neurons) in the SFO of MSG-treated rats of either sex and at two different ages. Our results shown that the treatment of neonatal rats with MSG induced a substantial reduction in the volume of the SFO and in the number of its nitrergic cells with regard to control animals. These findings suggest that the SFO could be implicated in some of the cardiovascular alterations observed in MSG treated rats. PMID- 15352880 TI - The macroanatomy of coronary arteries in donkeys (Equus asinus L.). AB - The aim of this study was to define the arteries that supply the cardiac muscle in donkeys with regard to their course and possible variations. Six hearts belonging to donkeys of different age and sex constituted the material of the study. Following exposition of the arteries by means of injection of latex coloured with Rotring ink to a. coronaria sinistra and a. coronaria dextra, dissection was performed. The arterial vascularization of the heart in donkeys was determined to be supplied by a. coronaria sinistra and a. coronaria dextra which originate from the aorta. A. coronaria sinistra, measured to be larger than a. coronaria dextra, was determined to have its origin at the aorta, at the level of the free border of valvula semilunaris sinistra, and to extend between truncus pulmonalis and auricula sinistra. The mentioned artery was detected to give off two branches, namely, ramus interventricularis paraconalis and ramus circumflexus sinister which extend in sulcus interventricularis paraconalis and sulcus coronarius, respectively. However, a. coronaria dextra was determined to have its origin at the beginning of the aorta, at the level of valvula semilunaris dextra, and to extend to margo ventricularis dexter between truncus pulmonalis and auricula dexter. This study has revealed ramus interventricularis subsinuosus and ramus circumflexus dexter to intercommunicate by means of anastomosis and to form a. coronaria dexter. Examination of material revealed the absence of anastomosis between r. circumflexus sinister and r. circumflexus dexter. Branches named rami septales, originating from ramus interventricularis paraconalis and ramus interventricularis subsinuosus were determined to supply septum interventriculare. Ramus proximalis atrii sinistri was determined to have its origin at ramus circumflexus sinister whereas ramus proximalis venriculi dextri was determined to stem from the beginning of a. coronaria dextra in the material of this study. However, examination of the cranial and caudal branches of ramus coni arteriosi, revealed the cranial branch to be ramus proximalis ventriculi dextri in a cadaver. PMID- 15352881 TI - Histological, histochemical and immunohistochemical study of the haemal nodes of the dromedary camel. AB - Haemal nodes are lymphoid organs found in various mammals and some birds. The structure of haemal nodes has been described in a number of species but not yet in the camel. Therefore, haemal nodes from 10 camels were studied histologically and tested for CD3, CD22, major histocompatibility complex (MHC) class II/DR, alpha-smooth muscle actin and for the demonstration of acid and alkaline phosphatases. The haemal nodes were of spherical or kidney shape with one or two hili and had a capsule and trabeculae of connective tissue and smooth muscles. The main parenchyma was composed of a cortex and a medulla. The cortex was formed from lymphoid follicles and diffuse interfollicular lymphocytes. The medulla consisted of lymphoid cords separated by medullary sinuses. The interfollicular lymphocytes and those in the medullary cord were CD3-positive. The lymphoid follicles showed CD22-positive cells. MHC class II/DR was expressed by most cells of the parenchyma. There were also subcapsular, peritrabecular and medullary blood sinuses. Afferent and efferent lymphatics and lymphatic sinuses were also found. Acid phosphatase-positive cells were localized mainly in the marginal, the interfollicular zone and in the medullary cord. Alkaline phosphatase positivity was observed in the endothelium of the sinuses and in the lymphoid follicles. The morphology of these organs in the camel allows a classification as haemolymph nodes and suggests involvement in blood and lymph filtration. PMID- 15352882 TI - scanning electron microscopy of lingual papillae in the common shrew, Sorex araneus, L. AB - The dorsal surface of the tongue of the adult common shrew (Sorex araneus L.) was examined by scanning electron microscopy. As in the other insectivores, three types of lingual papillae were observed: filiform, fungiform and vallate papillae. The filiform papillae represented the most numerous type of lingual papillae. The characteristic feature of the filiform papillae, covering the apex and corpus of the tongue, is the two processes tilted to the root of the tongue. The filiform papillae on the lingual apex are reduced in size and structure. Five to six fungiform papillae are placed symmetrically along the left and right border of the corpus of the tongue. Two large oval vallate papillae are located on the radix of the tongue. The posterior surface of the tongue in common shrew is covered with a smooth mucosa with the openings of the serous glands. PMID- 15352883 TI - An anatomical study of an electric organ and its nerve supply in the electric ray (Torpedinidae Narke japonica). AB - Despite anatomist's great interest in the electric organ (EO) of the electric ray, its detailed morphology remains unclear. In order to understand more completely the architecture of the EO and the branchial organ, it is necessary to examine detailed relationships regarding the origin, course and distribution of nerves innervating the EO. We thus carried out a macroscopic and microscopic anatomical study, focusing on issues, using the 18 sides of nine electric rays. The following results were obtained: (i) the EO was innervated exclusively by the facial, glossopharyngeal and vagus nerves; (ii) although these three cranial nerves consistently innervated the EO, variation in the number of the nerves innervating the EO was observed; (iii) cranial nerves innervated the EO in a segmental manner, at both entry and in the area of distribution. These results suggest that the EO of the electric ray might have differentiated from a non constant branchial muscle anlage but preserves the branchial segments in terms of the craniocaudal, dorsoventral and proximodistal axes. PMID- 15352884 TI - Changes of calbindin D-28k immunoreactivity in the hippocampus after adrenalectomy in the seizure sensitive gerbil. AB - Calbindin D-28k (CB), a calcium-binding protein, containing neurons in the hippocampus plays an important role in hippocampal excitability in epilepsy. In the present study, we investigated changes of CB immunoreactivity after adrenalectomy (ADX) in the hippocampus and dentate gyrus of the seizure sensitive gerbil, which is susceptible to seizure to identify roles of CB in epileptogenesis. The changes of the CB immunoreactivity after ADX were significant in the hippocampal CA1 region. By 24 h after ADX, CB-immunoreactive CA1 pyramidal cells and CB immunoreactivity increased. At this time, well-stained dendrites projected to the stratum radiatum. Thereafter, the CB immunoreactivity decreased time dependently by 96 h after ADX. In the dentate gyrus, the changes of CB-immunoreactive neurons were mainly observed in the granule cell layer. The number and immunoreactivity of CB-immunoreactive neurons was high at 24 h after ADX, thereafter, those decreased by 96 h after ADX. These results suggest that glucocorticoid has an important role in modulating the seizure activity and CB serves an inhibitory function, which regulates the seizure activity and output signals from the hippocampus. PMID- 15352885 TI - The scanning electron and light microscopic structure of bovine tactile hair. AB - Bovine tactile hairs in skin samples from the lateral side of the upper lip were examined using scanning electron and light microscopy. The root of these hairs has a variable length and is surrounded by a large sinus located between the internal and the external dermal sheath. With a prominent thickness, the external dermal sheath forms the external wall of the tactile hair and contains many nerves some of which extend into trabeculae. Trabeculae projecting from the internal dermal sheath and attaching to the external dermal sheath with two or more branches are present in the entire sinus. The trabeculae are interconnected by connective tissue sheets that support the integrity of the trabecular organization. The sinus surfaces as well as trabeculae are lined by endothelia. As a result, the bovine tactile hair is truly a cavernous type of tactile hair with a well organized microscopic anatomy. Thus, the bovine tactile hair most likely plays an important role in relatively immobile and insensitive bovine lips. PMID- 15352886 TI - Immunohistochemical study of intermediate filaments and neuroendocrine marker expression in leydig cells of laboratory rodents. AB - The aims of this study were to detect the expression of intermediate filaments and to verify the existence of marker substances for neuronal and neuroendocrine cells within the interstitial Leydig cells of laboratory rodent's testes, such as it has been described in other species. Adult male rats, mice, gerbils, Syrian hamsters and guinea-pigs were used and the localization of the different markers was achieved by the streptoavidin-peroxidase immunohistochemical method. The present study demonstrates in all rodents studied a similar pattern of localization in Leydig cells of intermediate filaments (vimentin, cytokeratin, neurofilament 200 kD and glial fibrillary acidic protein) and other marker substances (S-100, CgA, substance P and neurone-specific enolase), which are typical of neuroendocrine (APUD cells or paraneurones) and glial cells. The expression of these substances, related to neurotransmitters or neurohomones and other proteins characteristic of neuroendocrine cells, could suggest that it is a neural crest derived cell. Although this study provides more evidences about the immunoexpression of neuronal and glial markers in Leydig cells, this fact cannot be related directly to their embryological origin, because the current data support the hypothesis of a mesenchymal origin of the Leydig cells. PMID- 15352887 TI - Review article: exploration of the genetic aetiology of inflammatory bowel disease--implications for diagnosis and therapy. AB - Genomic technologies offer new approaches to the investigation of the aetiology and pathophysiology of inflammatory bowel disease. An important field relevant to inflammatory bowel disease therapy is the pharmacogenetic investigation of gene variations that may predict responses to certain medications in order to target these therapeutic interventions more precisely. To date, only about 12,000 of the estimated 30,000-50,000 human genes have been characterized. Therefore, the use of techniques for a global analysis of gene expression may allow the identification of new pathways or molecules in the therapeutic mechanisms of drugs. Recently, NOD2 has been identified as the first disease gene in inflammatory bowel disease. DLGS and OCTN-1 have been named as further disease genes. Although the detection of disease-associated variants has greatly advanced our understanding of the primary events that lead to the development of inflammatory bowel disease in a subgroup of patients with Crohn's disease, the implications of the findings for diagnostic and therapeutic algorithms are less clear. However, it appears that there is a clear association between certain subphenotypes of Crohn's disease and the disease-associated variants in the NOD2 gene. It can be anticipated that genomic findings will profoundly influence the future therapy of inflammatory bowel disease. PMID- 15352888 TI - Review article: the aetiopathogenesis of inflammatory bowel disease--immunology and repair mechanisms. AB - Although the aetiopathogenesis of Crohn's disease and ulcerative colitis, remains unsolved, current evidence indicates that defective T-cell apoptosis and impairment of intestinal epithelial barrier function play important roles in the pathogenesis of both conditions. Without appropriate control of T-cell proliferation and death during an immune response, an inappropriate accumulation of T cells and subsequent intestinal inflammation may occur. Differences in T cell responses between Crohn's disease and ulcerative colitis have been identified, with mucosal T-cell apoptosis being defective in Crohn's disease, but not in ulcerative colitis. Furthermore, cell cycling is considerably faster, with a vigorous clonal expansion, in Crohn's disease, whereas, in ulcerative colitis, T cells cycle normally, but have a remarkably reduced capacity to divide and expand. The elimination of excessive T cells therefore seems to be a reasonable approach to restore the gut to a physiological state or, at least, a controlled state of inflammation. The tumour necrosis factor-alpha blocker, infliximab, exerts its beneficial effects, at least in part, by the induction of apoptosis in lamina propria T cells and monocytes. In addition, repeated damage and injury of the intestinal surface is a hallmark of inflammatory bowel disease and may facilitate the entry of luminal antigens into the mammalian organism and the initiation and perpetuation of both nonspecific and specific immune responses. A better understanding of and enhancement of intestinal repair mechanisms may thus provide future approaches for the treatment of inflammatory bowel disease. PMID- 15352889 TI - Review article: gut flora and inflammatory bowel disease. AB - The pathogenesis of inflammatory bowel disease involves interactions between the host susceptibility, mucosal immunity and intestinal microflora. There is therefore great interest in the changes in the endogenous flora in inflammatory bowel disease patients and in the establishment of potential genetic variations in host responses to endogenous bacteria. In this review, we summarize the modifications in the various regional ecosystems in the gastrointestinal tract during inflammatory bowel disease (luminal bacteria in faeces or inside the gastrointestinal tract, bacteria in mucus and bacteria directly attached to the mucosa). Results were obtained following a 'candidate microorganism strategy' and, as is occurring increasingly frequently, following a 'full description strategy', which has progressed largely due to the development of culture independent techniques. The possibility of modifying the ecosystem using prebiotics or probiotics offers hope for new treatment developments, particularly in the prevention of relapse. PMID- 15352890 TI - Review article: colorectal carcinoma and inflammatory bowel disease. AB - The risk of colorectal cancer for any patient with ulcerative colitis is estimated to be 2% after 10 years, 8% after 20 years and 18% after 30 years of disease. The relative risk of colorectal cancer in Crohn's colitis is approximately 5.6 and should raise the same concerns as in ulcerative colitis. Risk factors for colorectal cancer include disease duration, early onset, extensive disease, primary sclerosing cholangitis and a family history of sporadic colorectal cancer. All patients should have a review colonoscopy 8-10 years after diagnosis to establish the extent of the disease. Surveillance should begin 8-10 years after disease onset for pancolitis and 15-20 years after disease onset for left-sided disease. Regular surveillance is recommended, with a screening interval every 3 years in the second decade of disease and annually by the fourth decade. Random biopsies should be taken at regular intervals with attention paid to dysplasia-associated lesions or masses, irregular plaques, villiform elevations, ulcers and strictures. Dysplasia is recognized as a premalignant condition, but the likelihood of progression to cancer is difficult to predict. High-grade dysplasia, confirmed by two expert gastrointestinal pathologists, is a strong indication for colectomy, as is low-grade dysplasia, although the diagnosis of low-grade dysplasia is unreliable. Surveillance programmes indicate that the overall 5-year survival rate is higher in surveyed patients, although patients still present with Dukes C cancers or disseminated malignancy. Surveillance has huge socioeconomic implications. As surveillance is not 100% effective, alternative ways of reducing the cancer risk with chemopreventive agents, such as aminosalicylates, are being considered. PMID- 15352891 TI - Review article: the chemoprevention of colorectal carcinoma. AB - This review aims to provide an insight into some of the common pathways involved in the development of colorectal cancer--so-called chromosomal instability and microsatellite instability. There is both clinical and molecular evidence to show that colorectal cancer development occurs over an extended period of time. Together with a knowledge of the molecular pathogenesis of colorectal cancer, this time window allows physicians to counteract cancer development, a strategy known as chemoprevention. In familial and sporadic colorectal cancer, aspirin, other nonsteroidal anti-inflammatory drugs and cyclo-oxygenase-2 inhibitors have been tested for their ability to reduce polyp and cancer development in clinical studies. In inflammatory bowel disease-related colorectal cancer, evidence from retrospective case-control studies shows that mesalazine (mesalamine) can reduce the development of dysplastic lesions or cancer. Although the mechanisms behind this are incompletely understood, general anti-inflammatory, oxygen scavenger and some pro-apoptotic and cyclo-oxygenase inhibitory activities of mesalazine are thought to be involved. New molecular evidence points to a direct DNA stabilizing effect of mesalazine, resulting in a significant reduction of spontaneous microsatellite mutations. PMID- 15352892 TI - Review article: joint involvement in inflammatory bowel disease. AB - Peripheral involvement of the joints, including pauciarticular, asymmetrical, transitory and migrating synovitis and enthesiopathy, is observed in 10-20% of affected inflammatory bowel disease patients. Recurrence is common and frequently coincides with a flare-up of intestinal disease. The true prevalence of axial involvement is less well established. Sacroiliitis is a hallmark of spondylitis, but is under-reported due to its insidious onset and sometimes asymptomatic nature. Radiographic evidence of sacroiliitis is present in about 20-25% of patients. Ankylosing spondylitis, as defined by the Rome criteria, is present in 3-10% of inflammatory bowel disease patients, and is thought to have a different genetic predisposition in these patients compared with 'classic' ankylosing spondylitis: whereas the human leucocyte antigen B27 phenotype is present in 90% of patients with 'classic' ankylosing spondylitis, the prevalence decreases to only 30% in patients with ankylosing spondylitis secondary to Crohn's disease. Polymorphisms involving CARD15 appear to be a possible genetic trigger: 78% of patients with Crohn's disease and symptomatic or asymptomatic sacroiliitis carry at least one mutation, compared with only 48% of control Crohn's disease patients. Moreover, in other forms of spondyloarthropathy, a similar association has been reported: 42% of patients with spondyloarthropathy and associated asymptomatic chronic gut inflammation, who are considered likely to develop Crohn's disease and ankylosing spondylitis, are carriers of at least one CARD15 mutation, compared with only 7% of patients with normal histology. In addition to genetic markers, clinical features support the relationship between gut and joint pathophysiology. In cases of spondyloarthropathy, a very rapid, substantial and sustained improvement in symptoms has been reported following treatment with infliximab, suggesting an essential role for tumour necrosis factor-alpha in spondyloarthropathy, similar to that observed in Crohn's disease. PMID- 15352893 TI - Review article: bone disease in inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) is associated with an increased incidence of osteoporosis. Osteoporosis with osteoporotic pain syndromes, fragility fractures and osteonecrosis accounts for significant morbidity and impacts negatively on the quality of life. It is generally agreed that there is a need to increase awareness for inflammatory bowel disease-associated osteoporosis. However, the best ways in which to identify at-risk patients, the epidemiology of fractures and an evidence-based rational prevention strategy remain to be established. The overall prevalence of IBD-associated osteoporosis is 15%, with higher rates seen in older and underweight subjects. The incidence of fractures is about 1 per 100 patient years, with fracture rates dramatically increasing with age. While old age is a significant risk factor, disease type (Crohn's disease or ulcerative colitis) is not related to osteoporosis risk. Corticosteroid use is a major variable influencing IBD-associated bone loss; however, it is difficult to separate the effects of corticosteroids from those of disease activity. The recommendations in inflammatory bowel disease are similar to those for postmenopausal osteoporosis, with emphasis on lifestyle modification, vitamin D (400-800 IE daily) and calcium (1000-1500 mg daily) supplementation and hormone replacement therapy (oestrogens/selective oestrogen receptor modulators in women, testosterone in hypogonadal men). Bisphosphonates have been approved for patients with osteoporosis (T-score < 2.5), osteoporotic fragility fractures and patients receiving continuous steroid medication. Data on the recently Food and Drug Administration-approved osteoanabolic substance parathyroid hormone and on osteoprotegerin are promising in terms of both steroid-induced and inflammation mediated osteoporosis, the key elements of inflammatory bowel disease-associated bone disease. PMID- 15352894 TI - Review article: skin complications associated with inflammatory bowel disease. AB - Cutaneous manifestations are well-recognized complications of Crohn's disease and ulcerative colitis. The incidence of these manifestations varies widely but, at the time of diagnosis, the mean incidence is around 10%. During the course of the disease, a great variety of skin lesions may develop, many of which are secondary to granulomatous cutaneous disease, reactive skin eruptions, nutritional deficiency and other associated conditions. The disorders that are directly related to the inflammatory process of Crohn's disease include perianal and peristomal ulcers and fistulae, metastatic Crohn's disease and oral granulomatous lesions. Histologically, the features are similar to those found in the inflamed bowel. These lesions usually respond to treatment of the underlying intestinal disease. The most common forms of reactive skin eruption are erythema nodosum and pyoderma gangrenosum. Certain subsets of patients are more susceptible to the development of erythema nodosum; in a previous report from our group, erythema nodosum was seen mainly in females, and in patients with colonic involvement and/or arthritis. This manifestation tends to occur during the first 2 years of the clinical course of the disease and may recur in approximately one-half of cases. Infliximab is highly effective in healing refractory lesions of erythema nodosum and pyoderma gangrenosum. Manifestations that are secondary to nutritional deficiency or associated conditions include acrodermatitis enteropathica, psoriasis and autoimmune disorders. For most of the cutaneous manifestations, the primary therapeutic target remains the bowel. Early aggressive therapy can minimize severe complications and maintenance treatment may prevent some devastating consequences. PMID- 15352895 TI - Review article: patients' fears and unmet needs in inflammatory bowel disease. AB - The management of chronic illness is becoming increasingly patient-centred. Although patients with inflammatory bowel disease have a normal life expectancy, most individuals experience an impact of inflammatory bowel disease on their daily lives as well as on their attitudes, fears and beliefs. Although currently available therapies for ulcerative colitis and Crohn's disease are becoming increasingly effective, there are still many unmet needs to address in this patient population. Physicians and patients' spouses or significant others underestimate the type and severity of problems reported by inflammatory bowel disease patients. Physical problems are frequently measured using disease activity indices. Emotional and social problems are reported using quality of life questionnaires and other specific measurement tools pertinent to the question of interest. Studies have indicated a poorer physical and emotional function in inflammatory bowel disease patients than in the general population. Effective therapies, both medical and surgical, produce significant improvements in the general and disease-specific quality of life. The quality of life is worse when the disease is more severe. Concomitant anxiety or depression appears to impair the quality of life even further. Common fears include the possibility of unanticipated flares, the need for surgery, poor energy levels and the side effects of medication. Recent studies have suggested that more prominent patient participation in management has the potential for greater patient satisfaction, better outcomes and more efficient health resource utilization. Thus, future studies should focus not only on ensuring the wider availability of effective therapies, but on increasing access to health care that is tailored to individuals--more structure for some and more independence and self-management for others, with health provider supervision. PMID- 15352896 TI - Review article: aminosalicylates in inflammatory bowel disease. AB - Aminosalicylate therapy for ulcerative colitis remains a foundational strategy for the induction and maintenance of remission for mild to moderate disease. Although it seems clear that topical mesalazine (mesalamine) is the most efficacious approach to distal ulcerative colitis, recent trials with orally delivered azo conjugates suggest that there may be an advantage over pH-released mesalazine as a first-line approach to active disease. No such comparisons are available for azo products and the prolonged-release formulation, Pentasa. However, recent meta-analyses have demonstrated that, although there is little difference in systemic exposure between marketed products, luminal concentrations may vary. In maintenance therapy, aminosalicylates remain the standard approach after aminosalicylate-induced remission. A number of gaps remain in the evidence base with regard to the optimal dosing of oral mesalazine as a maintenance agent, whether oral mesalazine can maintain remissions after rectal mesalazine induction, and the dose-response and efficacy of aminosalicylates after steroid- or ciclosporin-induced remissions. Although aminosalicylates have been advocated for several decades in Crohn's disease, a number of controversies have evolved since the original trials with sulfasalazine in active Crohn's disease. The original trials demonstrated benefits for sulfasalazine in colonic involvement, but controlled trial evidence for the role of sulfasalazine as maintenance therapy has not been as firmly established. In addition, although oral mesalazine has been demonstrated in controlled trials to be superior to placebo in mild to moderate disease, it is less efficacious than corticosteroids at inducing remissions. The maintenance benefits of mesalazine appear to be limited to patients 'induced into remission' with mesalazine and in some post-operative settings. PMID- 15352897 TI - Review article: systemic and topical steroids in inflammatory bowel disease. AB - Steroids are still widely used in the treatment of inflammatory bowel diseases. Pharmacological studies have shown that there is no major abnormality in the pharmacokinetics of steroids in these disorders. Foam preparations with rectal application decrease the bioavailability to low levels, eliminating systemic complications. For oral use, 'nonsystemic' steroids have been developed. In ulcerative colitis, steroids are rarely needed as 5-aminosalicylates are effective in the majority of patients. This is true for rectal application in distal colitis, as well as in more extensive disease. In Crohn's disease, steroids are more often used; however, in population-based studies, less than 50% of patients have been treated with steroids, as there are alternative treatments available for the large group of patients with mild to moderate activity. For those patients needing steroid treatment, budesonide seems to be a good choice in active disease, but has not shown convincing effects in the maintenance of remission over longer periods of time. There is no place for long-term steroid treatment in ulcerative colitis and very little in Crohn's disease- immunosuppression with azathioprine or related drugs is certainly the better alternative. PMID- 15352898 TI - Review article: antibiotics and probiotics in inflammatory bowel disease. AB - Treatment with antibiotics in inflammatory bowel disease has a long tradition and is widely used. The indications for antibiotic therapy are wide ranging, from specific situations such as abscesses or fistulae, to patients with severe disease (as an unspecific 'protective' measure), and to address the hypothesis that the enteric flora as a whole, or specific microorganisms such as mycobacteria, are involved in the pathogenesis of inflammatory bowel disease. The best-studied single antibiotic compound is metronidazole. However, overall, the scientific basis for the use of antibiotics is limited, which may reflect a lack of interest from sponsors within the pharmaceutical industry. Despite this weak evidence base, antibiotics are a globally established therapeutic tool in inflammatory bowel disease. Growing evidence from human and animal studies points towards a pivotal pathogenetic role of intestinal bacteria in inflammatory bowel disease. In view of these experimental findings, clinical trials have been undertaken to elucidate the therapeutic effects of probiotics in inflammatory bowel disease. Probiotics are viable nonpathogenic microorganisms which confer health benefits to the host by improving the microbial balance of the indigenous microflora. So far, of the many candidates, one specific strain (Escherichia coli Nissle 1917) and a mixture of eight different bacteria have demonstrated convincing therapeutic efficacy in controlled studies. Maintenance therapy in ulcerative colitis and prevention therapy, as well as the treatment of pouchitis, have emerged as areas in which probiotic therapy offers a valid therapeutic alternative to current treatments. Further investigations may detect additional clinically effective probiotics and other clinical indications. PMID- 15352899 TI - Review article: the role of nutrition in the treatment of inflammatory bowel disease. AB - Nutrients may be involved in the modulation of the immune response through at least three different mechanisms. First, the intestinal ecosystem plays a pivotal role in the pathogenesis of inflammatory bowel disease, triggering the uncontrolled inflammatory response in genetically predisposed individuals. Nutrients, together with bacteria, are major components of, and can therefore influence, the intestinal environment. Second, as components of cell membranes, nutrients can mediate the expression of proteins involved in the immune response, such as cytokines, adhesion molecules and nitric oxide synthase. The composition of lipids in the cell membrane is modified by dietary changes and can influence cellular responses. Indeed, various epidemiological, experimental and clinical data suggest that the immune response may be sensitive to changes in dietary composition. Finally, suboptimal levels of micronutrients are often found in both children and adults with inflammatory bowel disease, although, with the exception of iron and folate, it is unusual to discover symptoms attributable to these deficits. However, subclinical deficits may have a pathophysiological significance, as they may favour the self-perpetuation of the disease (due to defects in the mechanisms of tissue repair), cause defective defence against damage produced by oxygen free radicals and facilitate lipid peroxidation. These events can occur even in clinically inactive or mildly active disease, as well as in the development of dysplasia in the intestinal mucosa. Some dietary manipulations have been attempted as primary treatment for rheumatoid arthritis, and specially formulated diets for enteral nutrition have proved to be an effective treatment for Crohn's disease. Most trials, although lacking sufficient patient numbers, have demonstrated a role for dietary manipulation as primary therapy for inflammatory disease. Dietary lipids are one of the most active nutritional substrates modulating the immune response. Recently, it has been demonstrated that lipids may be a key factor explaining the therapeutic effect of clinical nutrition in Crohn's disease. PMID- 15352900 TI - Review article: inflammatory bowel disease--empowering the patient and improving outcome. AB - Inflammatory bowel disease, in common with most chronic diseases, is managed by specialist clinicians during regular clinic follow-up visits. Patients spend approximately 1 h per year with clinicians, with few provisions made for patient participation in their own management for the remainder of the year, resulting in dependence and disempowerment. The provision of regular, fixed clinic appointments for diseases that follow an unpredictable relapsing/remitting pattern results in inefficiencies for health services and inconvenience for patients, as well as high rates of noncompliance. Self-care is a normal human function and accounts for the management of three-quarters of all episodes of ill health. More formalized applications include patients and doctors working collaboratively to develop a set of guidelines which patients use to manage their chronic disease themselves. Recent studies have shown that, in patients with stable ulcerative colitis, self-management results in reduced health service utilization, speedier access to treatment and high levels of patient acceptability, without compromising health outcomes. There are a number of barriers to the effective implementation of guided self-management at many levels. Clinicians may be reluctant to pass control of treatment changes to patients, particularly the use of steroids. Access to clinics at short notice may be difficult and some patients themselves prefer a system in which all decisions are made by doctors. Research into guided self-management is ongoing as the long term outcomes are uncertain. However, there are indications that passing 'ownership of management' back to patients may improve compliance as patients realize their own responsibilities for remaining well. PMID- 15352901 TI - Review article: the management of mild to severe acute ulcerative colitis. AB - The goals for the management of acute ulcerative colitis are the objective evaluation of disease activity, induction of remission, prevention of relapse and treatment of complications. Clinical practice should be guided by simple activity indices, as it is easy to underestimate severity. For the induction of remission, topical treatment with mesalazine (mesalamine) is appropriate initial therapy for distal disease but, if symptoms persist for over a fortnight, decisive treatment is usually appreciated by the patient. For mild to moderate disease, corticosteroids have been the mainstay in Europe, although high-dose aminosalicylates (such as Pentasa, 4 g orally daily and 1 g rectally) are an alternative for symptoms not interfering with daily activity. Novel therapeutic approaches in ulcerative colitis have lagged behind those used for Crohn's disease, but several (epidermal growth factor, RDP 58, basiliximab, leucocytapheresis) are on the horizon. Severe colitis, defined as a bloody stool frequency of more than six per day with any one of tachycardia (pulse > 90 beats/min), temperature (> 37.8 degrees C), anaemia (haemoglobin < 10.5 g/dL) or raised erythrocyte sedimentation rate (> 30 mm/h), is an indication for intensive intravenous treatment. National UK figures indicate that 30% of ulcerative colitis cases progress to colectomy, and objective criteria for predicting the need for colectomy have been validated. The timing of colectomy is the most important decision that a physician is called upon to make, in conjunction with the patient and surgical colleagues. For the maintenance of remission, aminosalicylates continue to be first-line therapy, although the choice of 5 aminosalicylate appears to be influenced as much by geography as by theoretical considerations. Steroids have no place in the maintenance of remission. Indications for azathioprine include patients after a severe relapse of ulcerative colitis, those with early relapse after steroids (dose of < 15 mg/day, or within 6 weeks of stopping) and those needing a second course of steroids within a year. Therapeutic decisions should have a strategy, aimed at navigating the patient around relapses and through to sustained remission. Good management depends on clinical skills, compassion and care of the individual, in addition to pharmaceuticals. PMID- 15352902 TI - Review article: problematic proctitis and distal colitis. AB - About two-thirds of patients with ulcerative colitis have an inflammatory involvement distal to the splenic flexure, and therefore may be effectively treated with topical treatment, allowing the delivery of the active drug directly to the site of inflammation and limiting systemic absorption and potential side effects. Topical aminosalicylate therapy is the most effective approach, and most patients will benefit hugely, provided that the formulation reaches the upper extent of the disease. Therefore, the choice of topical preparation should be based on the proximal extent of the disease and on patient preference. Oral aminosalicylates are less effective than topical therapies; however, a combination of oral and topical aminosalicylates can be successful in refractory patients. Alternatives to aminosalicylates are the new glucocorticoids, budesonide and beclometasone dipropionate, either as enemas or oral formulations (only beclometasone dipropionate). A combination of oral or rectal new glucocorticoids with rectal aminosalicylates should be considered in patients refractory to either approach. When these measures fail, treatment with oral glucocorticoids is necessary. An intensive intravenous steroid regimen is also helpful for patients refractory to oral steroids. Alternative treatments include short-chain fatty acid enemas, nicotine enemas and patches, acetarsol suppositories, ciclosporin enemas and epidermal growth factor enemas. Several factors potentially having a negative impact on therapeutic response include concurrent enteric pathogens, coexistent irritable bowel syndrome, patient nonadherence to therapy, inadequate dosing and duration of therapy, and proximal progression of the disease. Surgical colectomy may be required in those rare patients refractory or intolerant to pharmacotherapy. PMID- 15352903 TI - Review article: the long-term management of ulcerative colitis. AB - After the induction of remission, the second priority of therapy for ulcerative colitis is sustained clinical remission, defined as the absence of inflammatory symptoms (diarrhoea, bleeding, rectal urgency) and the maintenance of an intact mucosa, with the absence of ulcers, friability or significant granularity at endoscopy. The 'optimal' maintenance strategy will depend on the therapy needed to induce remission. Thus, the transition from induction to maintenance therapy will be determined by the intensity of acute therapy necessary to induce remission and the duration of therapy required to complete the resolution of clinical symptoms. There are few controlled clinical trials pertaining to maintenance after each induction regimen. However, experience dictates that aminosalicylates are efficacious after aminosalicylate-induced remissions, that steroids should be tapered according to the time required to induce remission, that patients requiring ciclosporin will benefit from the addition of long-term immunomodulation with azathioprine or mercaptopurine, and that many patients with distal colitis who require topical mesalazine (mesalamine) will continue to need topical therapy to maintain remission, albeit at reduced frequency. The expectations for maintenance therapy require patient adherence to the prescribed treatment regimen. Patients require education with regard to the long-term goals of maintenance therapy (e.g. prevention of relapse, reduction of long-term complications of disease activity or risks of acute therapy with steroids), and should be warned against the use of nonsteroidal anti-inflammatory drugs and cautioned about the cessation of smoking, when applicable, due to potential risks of relapse or chronic activity. PMID- 15352904 TI - Review article: chronic active disease and maintaining remission in Crohn's disease. AB - The clinical management of Crohn's disease can be considered in relation to the treatment of acute disease and the maintenance of remission. The medication used to achieve these two goals may or may not be the same. Some patients with mildly active disease may respond to high-dose (4 g/day) mesalazine (mesalamine), and 5 aminosalicylic acid may also be helpful in weaning a patient off steroids after treatment for a flare-up. However, the value of 5-aminosalicylic acid in maintaining remission in Crohn's disease remains controversial. Subgroups of patients may be helped: for example, patients with Crohn's disease who have experienced a relapse within the last 2 years may benefit. Steroids form the first-line therapy for acute episodes of inflammation but do not maintain remission. Azathioprine and mercaptopurine are the first-line drugs for the maintenance of remission in moderate to severe Crohn's disease, and by titrating the dose up from 2 mg/kg daily, some previously resistant patients will be brought into remission. One-half of patients who do not tolerate azathioprine will tolerate mercaptopurine. Methotrexate is effective in inducing and maintaining remission, and is useful for patients who fail azathioprine treatment. Thalidomide is not proven in controlled studies, but two open studies have demonstrated its efficacy. The optimal dose, however, remains to be defined. Purified liquid diets with food exclusion can induce remission in patients with active disease, but food exclusion is difficult to maintain long term. Infliximab can induce and maintain remission in patients resistant to other therapies, with two-thirds of patients initially responding to treatment. One-third go into remission and, of those who respond to a single treatment, approximately one-half maintain remission when treated regularly for a year. Infliximab is, however, associated with an increased risk of infection, and its effect on cancer incidence is uncertain. The development of antibodies against the drug is associated with a loss of effect and allergic infusion reactions. In summary, simple proven therapies should be used first, because of their safety and benefit in some patients. However, aggressive therapy should be used when needed. PMID- 15352905 TI - Review article: treatment of perianal fistulizing Crohn's disease. AB - Fistulizing Crohn's disease can involve the bowel, but is more commonly seen in the perianal region. In acute perianal Crohn's disease, perianal lesions are manifestations of disease activity and are frequently treated concomitantly with bowel lesions. Spontaneous resolution occurs in up to 50% of patients. Fistulae are secondary lesions that may progress to destruction of the sphincter apparatus necessitating proctectomy after years of suffering. The control of sepsis is the first objective. The drainage of abscesses and the placement of setons are essential steps in treatment. Disease severity can be readily assessed by examination under anaesthesia and by magnetic resonance imaging. Endoscopic ultrasonography is sensitive, but is hampered by the necessary introduction of a large instrument into an often narrowed anorectum. Antibiotics, especially metronidazole and ciprofloxacin, are useful short-term therapies to decrease or stop drainage, but relapse is immediate on discontinuation. Immunosuppression with azathioprine (2.5 mg/kg per day) or mercaptopurine (1.5 mg/kg per day) is effective, but slow and often incomplete. The management of perianal fistulizing disease resistant to standard treatment has greatly improved with the introduction of the anti-tumour necrosis factor-alpha antibody, infliximab. The complete arrest of the drainage of fistulae is obtained in 46% of patients 10 weeks after the administration of 5-10 mg/kg of infliximab at weeks 0, 2 and 6 and, on average, lasts for 12 weeks. A treatment algorithm for fistulizing Crohn's disease must therefore involve the early and optimal use of immunosuppression and of infliximab. Medical and surgical co-operation is also critical to achieve the best possible outcome. PMID- 15352906 TI - Cost-effectiveness of thiopurine methyltransferase genotype screening in patients about to commence azathioprine therapy for treatment of inflammatory bowel disease. AB - BACKGROUND: Azathioprine is a useful agent in the management of inflammatory bowel disease. Its use is limited by its side-effect profile. Marrow toxicity occurs in approximately 3.2% of patients and is known to be associated with diminished thiopurine methyltransferase enzyme activity resulting from genetic polymorphisms. AIM: To evaluate the cost-effectiveness of screening for thiopurine methyltransferase gene polymorphisms prior to initiation of azathioprine therapy. METHODS: Analysis of the literature was undertaken to calculate the expected frequency of leucopenia and its relationship with thiopurine methyltransferase polymorphisms in a model of theoretical inflammatory bowel disease patients. Decision analysis was then applied to assess the cost of a pre-treatment genotyping strategy, taking account of direct costs and cost per life-year saved. RESULTS: In 1000 inflammatory bowel disease patients treated with azathioprine, 32 will develop myelosuppression and one will die because of this. Of those who develop myelosuppression during azathioprine therapy, 32% are attributable to lower thiopurine methyltransferase activity. Pre-treatment genotyping costs pound 347 per life-year saved for a 30 year old and pound 817 per life-year saved for a 60 year old. This compares favourably with other health care technologies. CONCLUSION: The use of pre-treatment screening for thiopurine methyltransferase polymorphisms in inflammatory bowel disease patients commencing azathioprine therapy represents good value for money. PMID- 15352907 TI - Azathioprine formulation optimizes metabolite profile in inflammatory bowel disease. AB - BACKGROUND: Recent studies have suggested that mercaptopurine metabolism is influenced by drug formulation (mercaptopurine vs. azathioprine) and concomitant use of 5-aminosalicylic acid medications. AIM: To determine the influence of dose, formulation and 5-aminosalicylic acid use on mercaptopurine metabolism. METHODS: Metabolites from 131 inflammatory bowel disease patients were analysed. Logistic regression was used to analyse correlations between dose and metabolite levels. Multivariate analysis was used to determine the effects of drug formulation and 5-aminosalicylic acid use. RESULTS: A positive correlation was detected between dose and 6-tioguanine nucleotides levels for azathioprine/Imuran formulation (P = 0.005) but not for mercaptopurine formulation. Adjusted mean 6 tioguanine nucleotides levels were similar for both formulations. Adjusted mean 6 methylmercaptopurine levels were higher for mercaptopurine formulation than for azathioprine formulation (1950 vs. 1056, P = 0.04). 5-Aminosalicylic acid use: 6 tioguanine nucleotides levels did not differ based on concomitant 5 aminosalicylic acid use. However, 5-aminosalicylic acid use did result in higher adjusted mean 6-methylmercaptopurine levels: 2078 on 5-aminosalicylic acid vs. 991 off 5-aminosalicylic acid (P = 0.004). CONCLUSIONS: (i) Azathioprine may have metabolic benefits by achieving a correlation of dose with 6-tioguanine nucleotides levels and by leading to lower mean 6-methylmercaptopurine levels. (ii) 5-aminosalicylic acid use does not significantly impact 6-tioguanine levels and may lead to higher 6-methylmercaptopurine levels. PMID- 15352908 TI - Rapid improvement of bone metabolism after infliximab treatment in Crohn's disease. AB - BACKGROUND: Crohn's disease is associated with low bone mineral density and altered bone metabolism. AIM: To assess the evolution of bone metabolism in Crohn's disease patients treated with infliximab. METHODS: We studied 71 Crohn's disease patients treated for the first time with infliximab for refractory Crohn's disease. Biochemical markers of bone formation (type-I procollagen N terminal propeptide, bone-specific alkaline phosphatase, osteocalcin) and of bone resorption (C-telopeptide of type-I collagen) were measured in the serum before and 8 weeks after infliximab therapy and compared with values in a matched healthy control group. RESULTS: Eight weeks after treatment with infliximab, a normalization of bone markers was observed with a median increase in formation markers of 14-51% according to marker and a lower but significant decrease in resorption marker (median 11%). A clinically relevant increase in bone formation markers was present in 30-61% of patients according to the marker. A clinically relevant decrease in C-telopeptide of type-I collagen was present in 38% of patients. No association was found with any tested demographic or clinical parameter. CONCLUSION: Infliximab therapy in Crohn's disease may rapidly influence bone metabolism by acting either on bone formation or bone resorption. This improvement seems to be independent of clinical response to infliximab. PMID- 15352909 TI - Diagnostic accuracy of faecal calprotectin estimation in prediction of abnormal small bowel radiology. AB - BACKGROUND: [corrected] Patients being investigated for symptoms of abdominal pain, diarrhoea and or weight loss often undergo small bowel radiology as part of their diagnostic workup mainly to exclude inflammatory bowel disease. AIM: To assess and compare the utility of a single faecal calprotectin estimation to barium follow through as well as conventional inflammatory markers such as erythrocyte sedimentation rate and C-reactive protein in exclusion of intestinal inflammation. METHODS: Seventy-three consecutive cases undergoing barium follow through for investigation of symptoms of diarrhoea and or abdominal pain with or without weight loss were studied. The control group comprised 25 cases with known active Crohn's disease (positive controls), 26 normal healthy volunteers (negative controls) and 25 cases of irritable bowel syndrome diagnosed by Rome II criteria. Symptoms, erythrocyte sedimentation rate and C-reactive protein were recorded at recruitment and a single stool sample assayed for calprotectin within 7 days prior to or after barium follow through. RESULTS: The median calprotectin value in the active Crohn's group, irritable bowel syndrome group and normal volunteers was 227 microg/g of stool, 19 and 10 microg/g respectively (P < 0.0001). A faecal calprotectin above a cut-off value of 60 microg/g was able to predict all nine cases with an abnormal barium follow through as well as all six cases with a normal barium follow through but with organic intestinal disease. The negative predictive value of a single calprotectin result below 60 microg/g of stool was 100% compared with 91% each for erythrocyte sedimentation rate > 10 mm and C-reactive protein > 6 mg/L and 84% for a combination of erythrocyte sedimentation rate and C-reactive protein in predicting absence of organic intestinal disease. CONCLUSION: A single stool calprotectin value < 60 microg/g of stool obviates the need for further barium radiology of the small bowel, is more accurate than measurement of erythrocyte sedimentation rate or C-reactive protein and effectively excludes Crohn's disease or non-functional gastrointestinal disease. PMID- 15352910 TI - A pilot study of orlistat treatment in obese, non-alcoholic steatohepatitis patients. AB - BACKGROUND: Treatment options for non-alcoholic steatohepatitis (NASH) are limited. Weight loss remains the most recommended therapy. Orlistat is an effective adjunct to dietary weight loss therapy. AIM: To evaluate the efficacy of orlistat, given for 6 months to patients with obesity and biopsy confirmed NASH. METHODS: Ten obese patients with biopsy proven NASH were enrolled. Orlistat was given with meals for 6 months. Body Mass Index (BMI), liver enzymes, haemoglobin A1c, fasting lipids and glucose were assessed at baseline and at completion of the study. Paired liver histology was obtained. RESULTS: Six women and four men were enrolled. The mean weight loss was 22.7 lb and ranged from 0 to 24.3%. The following clinical values significantly improved: mean BMI: 43.4-39.8 (P = 0.007); mean haemoglobin A1c (%): 7.14-5.95 (P = 0.021); mean alanine aminotransferase (ALT) (U/L): 93 -54 (P = 0.009); and mean aspartate aminotransferase (AST) (U/L): 79-48 (P = 0.008). Steatosis improved in six patients, and fibrosis improved in three patients. CONCLUSIONS: Orlistat therapy and dietary counselling were associated with significant decreases in body weight, haemoglobin A1c, ALT and AST. A 10% or greater reduction in weight improved steatosis and fibrosis as well as haemoglobin A1c levels in the majority of patients treated for 6 months. Controlled trials of longer duration are warranted to assess for histopathologic improvement as well as cost-efficacy in comparison to diet and exercise alone. PMID- 15352911 TI - Combination therapy for the treatment of hepatitis C in the veteran population: higher than expected rates of therapy discontinuation. AB - AIM: To compare the efficacy of high-dose induction with standard dose interferon therapy for the treatment of chronic hepatitis C virus at the Dallas Veterans Affairs Medical Center. METHODS: Patients were randomized to receive 5 million units daily interferon-alpha2b for 4-weeks followed by 44-weeks for genotype 1 or 20 weeks for non-genotype 1 of standard dose therapy (3 million units three times a week) or standard dose therapy for total treatment duration. Daily weight-based ribavirin was used for entire therapy interval. RESULTS: Forty-five patients were enrolled in the trial with genotype 1 comprising 75.6% of the sample. Cirrhosis or bridging-fibrosis was present in 69% of the patients. Of the 29 liver biopsies available for Knodell scoring, 41% and 51% had scores of 6-10 and 11-15, respectively. Rates of sustained virological response did not differ significantly between the two treatment groups. Therapy type and/or early intervention for depression did not affect the rate of therapy discontinuation, which was 26.6%. CONCLUSION: The rate of sustained virological response was similar between the two treatment groups and higher than anticipated among patients with cirrhosis or bridging-fibrosis. The rate of therapy discontinuation was also higher than anticipated but was not attributable to therapy type or untreated depression. PMID- 15352912 TI - Oesophageal and gastric pH profiles in patients with gastro-oesophageal reflux disease and Barrett's oesophagus treated with proton pump inhibitors. AB - BACKGROUND: Acid plays a significant role in the development of gastro oesophageal reflux symptoms and tissue damage. It is generally assumed that acid suppressive therapy with proton pump inhibitors improves or eliminates symptoms of gastro-oesophageal reflux disease by normalizing intra-oesophageal pH. However, the degree of acid suppression induced by proton pump inhibitor therapy in patients with gastro-oesophageal reflux disease and/or Barrett's oesophagus has not been adequately studied. AIM: To assess the efficacy of proton pump inhibitors in normalizing intra-oesophageal and intra-gastric pH in patients with gastro-oesophageal reflux disease with and without Barrett's oesophagus who have been rendered symptom-free by acid-suppressive therapy. METHODS: Patients with gastro-oesophageal reflux disease and Barrett's oesophagus were prospectively evaluated by dual sensor 24-h pH monitoring while receiving proton pump inhibitor therapy for complete control of gastro-oesophageal reflux disease symptoms. Analyses and comparisons of intra-oesophageal and intra-gastric pH profiles on therapy were then made. RESULTS: One hundred and ten patients, 98 men and 12 women, with gastro-oesophageal reflux disease (n = 62) and/or Barrett's oesophagus (n = 48), were studied. All tolerated proton pump inhibitors well and were asymptomatic at the time of the study. Thirty-six (58%) patients with gastro oesophageal reflux disease and 24 (50%) patients with Barrett's oesophagus (P = 0.4) normalized their intra-oesophageal pH profiles on proton pump inhibitors. Compared with patients with gastro-oesophageal reflux disease, patients with Barrett's oesophagus were more likely to have higher degree of pathologic acid reflux despite proton pump inhibitor therapy (DeMeester score 50.5 +/- 8.2 vs. 31.4 +/- 4.6, P = 0.03) and exhibited less intra-gastric acid suppression (% total pH < 4.0: 53.9 +/- 2.7 vs. 39.9 +/- 2.6, P = 0.0004), particularly supine (% pH < 4.0: 62.1 +/- 3.4 vs. 44.8 +/- 3.4, P = 0.0006). CONCLUSIONS: Gastro oesophageal reflux disease patients with or without Barrett's oesophagus continue to exhibit pathologic gastro-oesophageal reflux disease and low intra-gastric pH despite proton pump inhibitor therapy that accomplishes complete reflux symptom control. Further, intra-oesophageal and intra-gastric pH control is significantly more difficult to achieve in patients with Barrett's oesophagus. These findings may have significant therapeutic implications. PMID- 15352913 TI - Daily use of non-steroidal anti-inflammatory drugs is less frequent in patients with Barrett's oesophagus who develop an oesophageal adenocarcinoma. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs use may protect against development of oesophageal adenocarcinoma. AIM: To define the consequences of non steroidal anti-inflammatory drugs use in patients with Barrett's oesophagus. METHODS: Records of all Barrett's oesophagus/oesophageal adenocarcinoma patients examined in Blackpool-Wyre-Fylde area were reviewed. All surviving patients completed validated questionnaires. RESULTS: Use of non-steroidal anti inflammatory drugs of any type and at any frequency was more prevalent in Barrett's oesophagus patients [147 (38%) Barrett's oesophagus vs. 30 (26%) oesophageal adenocarcinoma, P = 0.02]. Daily use of non-steroidal anti inflammatory drugs was more prevalent in Barrett's oesophagus patients [88 (23%) Barrett's oesophagus vs. 14 (12%) oesophageal adenocarcinoma, P = 0.02], due to more prevalent consumption of non-aspirin non-steroidal anti-inflammatory drugs [48 (13%) Barrett's oesophagus vs. four (4%) oesophageal adenocarcinoma, P = 0.009]. There was no difference between the two groups in usage of either daily low-dose aspirin or of occasional non-steroidal anti-inflammatory drugs. In logistic regression analysis any use of non-steroidal anti-inflammatory drugs [odds ratio (OR) = 0571 (95% CI: 0.359-0.909), P = 0.018] and daily use of non aspirin non-steroidal anti-inflammatory drugs [OR = 0.297 (95% CI: 0.097-0.911), P = 0.034] were significant protective factors. Non-steroidal anti-inflammatory drugs use did not affect the survival of oesophageal adenocarcinoma patients. Oesophageal adenocarcinoma and Barrett's oesophagus consuming non-steroidal anti inflammatory drugs did not differ in upper gastrointestinal bleeding [26 (15%) non-steroidal anti-inflammatory drugs consumers vs. 29 (9%) non-consumers, P = 0.08], oesophageal ulcers [31 (18%) non-steroidal anti-inflammatory drug consumers vs. 49 (15%) non-consumers, P = 0.43] or stricturing [19 (11%) non steroidal anti-inflammatory drug consumers vs. 41 (13%) non-consumers, P = 0.58]. CONCLUSIONS: (i) Daily use of non-steroidal anti-inflammatory drugs is more prevalent in Barrett's oesophagus than oesophageal adenocarcinoma patients, because of a more prevalent use of non-aspirin non-steroidal anti-inflammatory drugs. (ii) Use of non-steroidal anti-inflammatory drugs in Barrett's oesophagus patients is safe if acid suppression is adequate. PMID- 15352914 TI - Esomeprazole 20 mg on-demand is more acceptable to patients than continuous lansoprazole 15 mg in the long-term maintenance of endoscopy-negative gastro oesophageal reflux patients: the COMMAND Study. AB - BACKGROUND: Symptom relief, through adherence to appropriate maintenance therapy, is the sole objective of treatment for patients with endoscopy-negative gastro oesophageal reflux disease. AIM: To compare the efficacy of 'on-demand' treatment with esomeprazole 20 mg vs. continuous treatment with lansoprazole 15 mg daily in patients with endoscopy-negative gastro-oesophageal reflux disease. METHODS: Endoscopy-negative gastro-oesophageal reflux disease patients who achieved complete resolution of heartburn after short-term (2-4 weeks) treatment with esomeprazole 20 mg (n = 774) were randomized to receive either esomeprazole 20 mg on-demand (n =311) or lansoprazole 15 mg continuous daily treatment (n = 311) for 6 months. RESULTS: Significantly more patients were willing to continue taking esomeprazole on-demand than lansoprazole continuous therapy after 6 months (93% vs. 88%; P = 0.02). This superior outcome was achieved despite patients on esomeprazole requiring medication only 38% as often as those on lansoprazole, leading to direct cost savings of more than one-third (36%). Furthermore, patients receiving esomeprazole 20 mg on-demand were more satisfied with their treatment after 1 month compared with patients taking lansoprazole 15 mg continuously. CONCLUSIONS: In patients with endoscopy-negative gastro-oesophageal reflux disease, esomeprazole 20 mg on-demand is more acceptable to patients and is an economically more effective treatment than lansoprazole 15 mg continuously. PMID- 15352915 TI - Long-term outcome and objective changes of anorectal function after biofeedback therapy for faecal incontinence. AB - BACKGROUND: In the short-term, biofeedback therapy improves symptoms and anorectal function in patients with faecal incontinence but whether there is long term improvement is incompletely understood. AIM: To prospectively evaluate bowel symptoms and anorectal function, both immediately and 12 months after biofeedback therapy. METHODS: A total of 105 consecutive patients (male/female = 12/93) with faecal incontinence, unresponsive to supervised medical treatment were enrolled in biofeedback training consisting of biweekly pelvic muscle strengthening exercises, anal squeeze and sensory-motor coordination training, and reinforcement sessions at 3, 6 and 12 months. Anorectal manometry, saline continence test, stool diaries and bowel satisfaction scores were used to assess improvement. RESULTS: 94/105 (male/female = 10/84) completed training and 11 dropped out. Sixty completed 1-year assessment. At 1-year, 63% reported no episodes of incontinence. Biofeedback decreased (P < 0.001) stool frequency and number of incontinence episodes and increased (P < 0.001) bowel satisfaction score, anal resting and squeeze pressures, squeeze duration and ability to retain saline infusion increased (P < 0.001), both immediately and at 1 year. Sensory thresholds decreased (P < 0.001). CONCLUSIONS: Biofeedback therapy produced sustained improvement in bowel symptoms and anorectal function. Because it is safe, inexpensive compared with other surgical interventions, and effective, biofeedback should be offered to incontinent patients unresponsive to medical therapy. PMID- 15352916 TI - Effect of cyclo-oxygenase inhibitors on Helicobacter pylori susceptibility to metronidazole and clarithromycin. AB - BACKGROUND: We previously reported that aspirin inhibited Helicobacter pylori growth and suppressed the mutagenic effect of metronidazole. AIM: To determine the effects of a cyclo-oxygenase (COX)-2-specific inhibitor, SC-236, and a non selective COX inhibitor, indometacin, on the growth, urease activity and antimicrobial susceptibility of H. pylori. METHODS: Three H. pylori reference strains, and 18 clinical isolates were treated with SC-236 or indometacin for 24 and 48 h. Growth, urease activity and susceptibility to clarithromycin and metronidazole of the bacteria were assessed by viable colony counting, spectrophotometry and E-test respectively. RESULTS: SC-236 and indometacin inhibited H. pylori growth in a dose-dependent manner with the lowest inhibitory concentrations of 0.03 and 0.1 mm, and the lethal concentrations of 0.09 and 0.3 mm, respectively. The numbers of CFU/mL in Brucella broth containing 0.09 mm SC 236 were 2 log lower at 24 h, and even 3 log lower at 48 h than that at 0 h (P = 0.035, compared with the vehicle control). Treatment of 0.3 mm indometacin reduced the number of CFU/mL by 1 log at 24 h compared with that at 0 h (P = 0.037 compared with the vehicle control). Helicobacter pylori urease activity began to decrease with 0.06 mm SC-236 at 24 h (P = 0.016), and 0.3 mm indometacin at 48 h (P = 0.025). MICs of metronidazole and clarithromycin against H. pylori were decreased significantly in the presence of 0.03 mm SC-236 or 0.1 mm indometacin (all P < 0.001). CONCLUSION: Both SC-236 and indometacin suppressed the growth and urease activity of H. pylori in a dose-dependent manner, and increased its susceptibility to the antibiotics. PMID- 15352917 TI - Effects of effervescent ranitidine on gastric pH: comparison with almagate and placebo in fasting and postprandial conditions. AB - AIM: To compare the effect of effervescent ranitidine, almagate (magnesium carbonate-aluminium hydroxide) and placebo on gastric pH, in fasting and postprandial conditions. METHODS: Twelve healthy volunteers underwent a gastro oesophageal pH monitoring on three different occasions after the administration of each of the following randomly allocated treatments: almagate, effervescent ranitidine and placebo. Treatment effects were assessed in fasting and postprandial conditions. Onset and duration of alkalinization, percentage of time with pH > 4 and median gastric pH after treatments were calculated in both periods. RESULTS: Onset of action of effervescent ranitidine was similar to almagate in fasting [median 1.2 min (IQR: 0.6-12.7) vs. 2.9 min (0.4-227.6)] and postprandial conditions [1.4 min (0.5-4.9) vs. 4.1 min (1.3-63.8)] and both were significantly faster than placebo [fasting 211.1 min (2.7-240); postprandial 240 min (175.6-240)]. The duration of action of effervescent ranitidine was statistically significant longer than almagate in fasting [235 min (105.2-239.4) vs. 19.4 min (6.7-38.8)] and postprandial conditions [171.8 min (133.2-239.5) vs. 61.3 min (44.7-91.9)]. Effervescent ranitidine was more effective than almagate in increasing the percentage of time with a pH > 4 both in fasting (73.9% vs. 7.3%) and postprandial (59.1% vs. 21.3%). CONCLUSIONS: Effervescent ranitidine shows an effect on gastric pH as fast as almagate but provides a duration of alkalinization longer than almagate, in both fasting and postprandial conditions. PMID- 15352918 TI - Differential effects of two types of antidepressants, amitriptyline and fluoxetine, on anorectal motility and visceral perception. AB - BACKGROUND: Although antidepressants are used for functional gastrointestinal disorders, the mechanisms of their effects on gut are incompletely understood. AIM: To assess the effects of two types of antidepressants (tricyclic, serotoninergic) on anorectal motility and visceral perception. METHODS: A placebo controlled, randomized, double-blind, crossover study was performed in 12 healthy male volunteers who received a single oral dose of amitriptyline (80 mg), fluoxetine (40 mg) or placebo. Drug effects were assessed using phasic isobaric distensions of the rectum with an electronic barostat (11 levels from 1 to 51 mmHg) 4 h after drug intake. Maximal rectal volume and pressure, mean and residual pressures at upper anal canal, mean pressure at lower anal canal, defecation sensation (5-level scale) and visceral perception (visual analogue scale) were recorded at each level of distending pressure. RESULTS: Ten subjects completed the study. Compared with placebo, neither amitriptyline nor fluoxetine modified rectal compliance or visceral perception. Compared with placebo, antidepressants significantly reduced mean and residual pressures at upper anal canal (-18%, P = 0.0019, and -27%, P = 0.0002, respectively, for amitriptyline; 26%, P = 0.0001, and -33%, P = 0.0001, respectively, for fluoxetine) whereas only amitriptyline significantly reduced mean pressure at lower anal canal (-16%, P = 0.0008). CONCLUSION: Both antidepressants similarly relaxed the internal anal sphincter, probably through a non-specific mechanism, without modifying visceral perception. Only amitriptyline relaxed the external anal sphincter. PMID- 15352919 TI - Epidemiology of ischaemic colitis. PMID- 15352924 TI - A successful antipsychotic combination trial. Quo Vadis? PMID- 15352925 TI - Hypofrontality in schizophrenia: a meta-analysis of functional imaging studies. AB - OBJECTIVE: Hypofrontality is not a well-replicated finding in schizophrenia either at rest or under conditions of task activation. METHOD: Studies comparing whole brain and frontal blood flow/metabolism in schizophrenic patients and normal controls were pooled. Voxel-based studies were also combined to examine the pattern of prefrontal activation in schizophrenia. RESULTS: Whole brain flow/metabolism was reduced in schizophrenia to only a small extent. Resting and activation frontal flow/metabolism were both reduced with a medium effect size. Duration of illness significantly moderated resting hypofrontality, but the moderating effects of neuroleptic treatment were consistent with an influence on global flow/metabolism only. Pooling of voxel-based studies did not suggest an abnormal pattern of activation in schizophrenia. CONCLUSION: Meta-analysis supports resting hypofrontality in schizophrenia. Task-activated hypofrontality is also supported, but there is little from voxel-based studies to suggest that this is associated with an altered pattern of regional functional architecture. PMID- 15352926 TI - Poor maternal care and high maternal body mass index in pregnancy as a risk factor for schizophrenia in offspring. AB - OBJECTIVE: We investigated whether antenatal factors in mothers would increase the risk of schizophrenia in the offspring, and also examined any relationship between these factors and histories of obstetric complications (OCs). METHOD: Using the Mother and Child Health Handbooks of 52 patients with schizophrenia and 284 healthy subjects, we evaluated the risk-increasing effects of the frequency of antenatal care visits and mothers' body mass index (BMI) at both early and late pregnancy. RESULTS: In logistic regression analysis, there was a significant association between the number of antenatal care visits and the risk of the disorder; an increase in a unit of visits corresponds to a reduction of the risk by 12%. We also found a 24% increase in the risk with a one-unit increase of BMI at the early pregnancy, and a 19% increase at the late pregnancy. These antenatal factors were found to contribute, in part, to an excess of OCs in individuals with schizophrenia. CONCLUSION: Poor maternal care during pregnancy and comparatively high maternal BMI especially at early pregnancy may cause a predisposition to schizophrenia in the offspring. PMID- 15352927 TI - Hallucinatory experiences and onset of psychotic disorder: evidence that the risk is mediated by delusion formation. AB - OBJECTIVE: To examine the hypothesis that the risk for onset of psychotic disorder in individuals with self-reported hallucinatory experiences (HE) would be higher in those who developed delusional ideation (DE) than in those who did not. METHOD: A population sample of 4673 individuals were interviewed with the Composite International Diagnostic Interview at baseline and 1 and 3 years later. At year 3, clinical re-interview took place to identify onset of psychotic disorder. RESULTS: Given the presence of HEs at baseline, the increase in risk of having the psychosis outcome at year 3 was much higher in those with DE at year 1 than in those without DE (risk difference between individuals with and without DE: 18.72%, 95% CI: 2.22-35.23, chi(2) = 4.94, df = 1, P = 0.026). CONCLUSION: The results are in line with current psychological theories stating that clinical outcome of psychosis-like experiences is related to the development of secondary beliefs and appraisals. PMID- 15352928 TI - Pleasurable auditory hallucinations. AB - OBJECTIVE: The focus in auditory hallucination (AH) research is usually on the negative impact of the experience itself. There are practically no studies on whether voices can be perceived as pleasurable. The aim of the present study was to assess the frequency of voices as a pleasurable experience in a psychotic patient population. METHOD: A total of 160 patients with AHs (89 schizophrenia and 17 other psychoses) were assessed with the psychotic symptom rating scale (PSYRATS) for AHs, including an added item on whether the experience was pleasurable. RESULTS: Twenty-eight patients (26%) reported the voices as a pleasurable experience and 10 of them did so frequently. Pleasurable hallucinations showed negative associations with amount and intensity of distress, degree of negative content and loudness. Positive associations were apparent with chronicity and perceived control over the voices. CONCLUSION: Pleasurable hallucinations can be detected in a substantial proportion of patients, and cross validated with existing instruments. PMID- 15352929 TI - Serum lipids in schizophrenia and other functional psychoses: a general population northern Finland 1966 birth cohort survey. AB - OBJECTIVE: To compare fasting serum lipid concentrations of subjects with schizophrenia with a comparison group. METHOD: The study sample consists of 5654 members of the northern Finland 1966 birth cohort who participated in the field study with blood samples after overnight fasting and clinical examination in 1997 98. Total cholesterol (TC), high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides (TG) and glucose were analyzed. Analysis of variance were used for comparing differences in lipids means between diagnostic categories. RESULTS: Mean fasting TC in subjects with schizophrenia was 20 mg/dl higher than in the comparison group. TC and TG levels in the group of other psychoses resembled the schizophrenia group. CONCLUSION: Blood lipid levels in subjects with schizophrenia and other functional psychoses were high. As these persons are at special risk for hyperlipidemia their lipid levels should be regularly monitored, and cholesterol lowering diet, as well as medication, should be considered. PMID- 15352930 TI - Does duration of untreated psychosis bias study samples of first-episode psychosis? AB - OBJECTIVE: While findings are contradictory, many studies report that long Duration of Untreated Psychosis (DUP) correlates with poorer outcome in first episode psychosis. In an outcome study of first-episode psychosis, we compared the patients who refused to participate in a follow-along with those who consented to estimate the importance of this factor in sample recruitment bias. Our questions were: (i) What is the percentage of refusers? (ii) Are there systematic differences between refusers and consenters on DUP and/or other admission variables? (iii) What is the risk of refusal for different values of DUP? METHOD: In an unselected group of consecutively admitted patients we compared follow-along refusers and consenters on the following admission variables: sex, age, diagnostic group, substance abuse, being in-patient, coming from an early detection site and DUP. We conducted a logistic regression analysis with refusal as the outcome variable. RESULTS: Ninety-three of 397 patients (23%) were refusers. In univariate analyses the only significant difference was found for DUP. The median DUP for consenters was 10 weeks and for refusers 32 weeks. DUP remained significant when all independent variables were controlled for. CONCLUSION: DUP was significantly related to risk for refusal. For tests of the impact of DUP on outcome, this may introduce a type II error bias. PMID- 15352931 TI - Amisulpride augmentation of clozapine: an open non-randomized study in patients with schizophrenia partially responsive to clozapine. AB - OBJECTIVE: Treatment options are very limited for individuals with schizophrenia resistant to clozapine. We tested the hypothesis that amisulpride augmentation would lead to an improvement in these patients. METHOD: This was an open non randomized study. Thirty-three patients with sub-optimal response to clozapine were commenced on amisulpride in addition to clozapine. Clinical status was evaluated at baseline, 3 and 6 months using the Positive And Negative Syndrome Scale (PANSS), Scale for the Assessment of Negative Symptoms (SANS), Global Assessment Scale (GAS), Calgary Depression Scale, Calgary Anxiety Scale and various side effect rating scales. RESULTS: Twenty-eight subjects completed 6 months treatment on clozapine and amisulpride. There was a statistically significant improvement in the mean scores for PANSS, SANS and GAS at follow-up and no significant changes in side effect ratings. CONCLUSION: Co-administration of amisulpride, in a group of patients partially or non-responsive to clozapine, may lead to a substantial improvement in positive and negative symptoms, without worsening the side effect burden. PMID- 15352932 TI - Obesity, dyslipidaemias and smoking in an inpatient population treated with antipsychotic drugs. AB - OBJECTIVE: The prevalence of cardiovascular disease in people with schizophrenia is known to be above population norms. In addition, many antipsychotic drugs have been associated with weight gain and some with dyslipidaemia. Our aims were to determine the apparent and actual prevalence of the modifiable risk factors for cardiovascular disease: obesity, hyperlipidaemia and smoking, in hospitalized patients treated with antipsychotic drugs. METHOD: (i) Survey of clinical notes. (ii) Direct measurement of fasting lipids, body mass index and waist : hip ratio. RESULTS: Lipids were rarely monitored and body weight often not recorded making the apparent prevalence of these cardiovascular risk factors low. Smoking status was recorded. When directly measured, the actual prevalence of obesity was above population norms and two-thirds of patients had a dyslipidaemia. CONCLUSION: Routine screening for these modifiable risk factors followed by active intervention where appropriate could significantly reduce morbidity and mortality in this patient group. PMID- 15352933 TI - The dental health of people with schizophrenia. AB - OBJECTIVE: To examine the dental health of community dwelling people with schizophrenia and to compare results with those in the general population. METHOD: Dental health of 428 people with schizophrenia in six different areas of the UK was assessed by a self-report questionnaire. RESULTS: Compared with the general population, significantly more of the younger patients were edentate (3 39% vs. 1-20%) and fewer had more than 20 teeth (70% vs. 83%). None of four dental health targets had been achieved in the patient population. More patients had last visited the dentist because of trouble with their teeth; fewer had visited for a check-up. Fewer patients cleaned their teeth daily; this group had more negative symptoms. CONCLUSION: The dental health of people with schizophrenia is poor. Community mental health teams should encourage them to attend their community dentist regularly. PMID- 15352934 TI - On the threshold of subthreshold depression. PMID- 15352937 TI - Amphetamine-like stimulant cessation in an abusing patient treated with buproprion (Tardiev et al. 2004). PMID- 15352950 TI - Echocardiography for cardiopulmonary optimization in the intensive care unit: should we expand its use? PMID- 15352951 TI - Troponin T-values provide long-term prognosis in elderly patients undergoing non cardiac surgery. AB - BACKGROUND: The aim of this study was to evaluate the significance of elevated postoperative Troponin T (TnT) levels in an elderly population undergoing non cardiac surgery. METHODS: Five hundred and forty-six consecutive patients aged 70 years or older undergoing non-cardiac surgery of >30-min duration were enrolled in this prospective, observational study. A postoperative TnT measurement was obtained on the 5th to 7th postoperative day. Troponin T values greater than 0.02 ng ml(-1) were considered positive. Patients were followed over a 1-year period, and mortality and non-fatal cardiac events (acute myocardial infarction and coronary interventions) were recorded. RESULTS: Troponin T concentrations greater than 0.02 ng ml(-1) were detected in 53 of the study subjects (9.7%). Eleven per cent of the patients with elevated TnT had electrocardiographic or clinical signs of myocardial ischemia. One year after surgery, 17 (32%) of the patients with abnormal TnT concentrations had died. In a multivariate Cox regression analysis adjusting for baseline and perioperative data, a TnT value >0.02 ng ml(-1) was an independent correlate of the mortality adjusted hazard ratio (HR): 14.9 (95% CI 3.7-60.3). Other independent predictors of death were tachycardia (HR, 14.9 95% CI 3.45-64.8), ASA 4 (HR, 8.1 95% CI 1.3-50.0), reoperation (HR, 6.4 95% CI 1.1 36.9), and use of diuretics (HR, 4.2 95% CI 1.3-13.8). CONCLUSION: We conclude that elevated TnT levels in the postoperative period confer a 15-fold increase in mortality during the first year after surgery. Our findings also provide evidence that silent myocardial ischemia is common in an elderly population. Routine perioperative surveillance for TnT might therefore be of use in detecting patients at an increased risk of mortality during the first postoperative year. PMID- 15352952 TI - Expiratory flow limitation in morbidly obese postoperative mechanically ventilated patients. AB - Although obesity promotes tidal expiratory flow limitation (EFL), with concurrent dynamic hyperinflation (DH), intrinsic PEEP (PEEPi) and risk of low lung volume injury, the prevalence and magnitude of EFL, DH and PEEPi have not yet been studied in mechanically ventilated morbidly obese subjects. In 15 postoperative mechanically ventilated morbidly obese subjects, we assessed the prevalence of EFL [using the negative expiratory pressure (NEP) technique], PEEPi, DH, respiratory mechanics, arterial oxygenation and PEEPi inequality index as well as the levels of PEEP required to abolish EFL. In supine position at zero PEEP, 10 patients exhibited EFL with a significantly higher PEEPi and DH and a significantly lower PEEPi inequality index than found in the five non-EFL (NEFL) subjects. Impaired gas exchange was found in all cases without significant differences between the EFL and NEFL subjects. Application of 7.5 +/- 2.5 cm H2O of PEEP (range: 4-16) abolished EFL with a reduction of PEEPi and DH and an increase in FRC and the PEEPi inequality index but no significant effect on gas exchange. The present study indicates that: (a) on zero PEEP, EFL is present in most postoperative mechanically ventilated morbidly obese subjects; (b) EFL (and concurrent risk of low lung volume injury) is abolished with appropriate levels of PEEP; and (c) impaired gas exchange is common in these patients, probably mainly due to atelectasis. PMID- 15352953 TI - Predictable reduction of intracranial hypertension with hypertonic saline hydroxyethyl starch: a prospective clinical trial in critically ill patients with subarachnoid haemorrhage. AB - BACKGROUND: After head trauma, hypertonic saline lowers intracranial pressure (ICP) and preserves or increases cerebral perfusion pressure (CPP). Hypertonic saline has not been studied in patients with increased ICP due to subarachnoid haemorrhage (SAH). The aim of this study was to evaluate the effects on elevated ICP and on CPP in patients critically ill from SAH. METHODS: Critically ill SAH patients needing urgent treatment for an elevated ICP, but otherwise stable, were included in this study. We infused 7.2% saline in 6% hydroxyethyl starch (HyperHAES((R)) Fresenius Kabi AG, Bad Homburg v.d.h., Germany) 2 ml kg(-1) during 20 min in 10 episodes of ICP > 20 mmHg in seven patients with SAH. Our primary outcome variables were changes in ICP and CPP during and for 3 h after this infusion. RESULTS: All interventions resulted in decreased ICP and elevation of CPP. The mean value for maximum ICP decrease in percent of baseline was 58% (range 43-83%, P = 0.002), which occurred at mean 40 min (range 25-90 min) after start of infusion. The mean percent peak increase in CPP was 26% (range 16-32%, P = 0.002). After 210 min, ICP was 35% lower than baseline (range 19-39%, P = 0.008). Serum sodium increase was mean 6.6 mmol l(-1) (range 5-9 mmol l(-1)) 30 min after start of infusion. CONCLUSIONS: 7.2% saline in 6% hydroxyethyl starch is an effective and safe therapy for intracranial hypertension after SAH. We demonstrate that an infusion of 2 ml kg(-1) during 20 min has a predictable and clinically significant beneficial effect on ICP and CPP. The effect was still present 3 h after end of infusion. Rebound ICP-increase was not observed within 3 h. PMID- 15352954 TI - Impact of a critical care outreach team on critical care readmissions and mortality. AB - AIMS: The aim of a critical care outreach team is to facilitate discharges from critical care beds, educate ward staff in the management of deteriorating patients, facilitate transfer to critical care and reduce readmission rates to critical care. Although intuitively a good idea, there are few data to support outreach in terms of reducing the readmission rate to critical care and subsequent patient mortality. This retrospective observational study attempted to determine the change in the critical care readmission rate, an indicator of the quality of critical care, critical care mortality and in-hospital mortality following the introduction of a critical care outreach team in a major teaching hospital. METHODS: A retrospective review of 1380 discharges from critical care was undertaken and the readmissions identified (n = 176). Readmission rate, mortality and other demographic data were compared between the pre and post outreach periods. RESULTS: Critical care mortality, in-hospital mortality and 30 day mortality were all reduced in the post-outreach period amongst readmissions to critical care. There was also a decease in the overall mortality of all patients admitted to critical care. There were no apparent causative factors for this reduction in mortality before and following outreach. CONCLUSIONS: There are many confounding factors in assessing the impact of outreach teams in hospitals. This study tentatively concludes that outreach teams may have a favourable impact on mortality rate amongst readmissions to critical care, but more data is needed from multicentre trials. PMID- 15352955 TI - Peripheral venous pressure is an alternative to central venous pressure in paediatric surgery patients. AB - BACKGROUND: Peripheral venous pressure (PVP) is easily and safely measured. In adults, PVP correlates closely with central venous pressure (CVP) during major non-cardiac surgery. The objective of this study was to evaluate the agreement between CVP and PVP in children during major surgery and during recovery. METHODS: Fifty patients aged 3-9 years, scheduled for major elective surgery, each underwent simultaneous measurements of CVP and PVP at random points during controlled ventilation intraoperatively (six readings) and during spontaneous ventilation in the post-anaesthesia care unit (three readings). In a subset of four patients, measurements were taken during periods of hypotension and subsequent fluid resuscitation (15 readings from each patient). RESULTS: Peripheral venous pressure was closely correlated to CVP intraoperatively, during controlled ventilation (r=0.93), with a bias of 1.92 (0.47) mmHg (95% confidence interval = 2.16-1.68). In the post-anaesthesia care unit, during spontaneous ventilation, PVP correlated strongly with CVP (r = 0.89), with a bias of 2.45 (0.57) mmHg (95% confidence interval = 2.73-2.17). During periods of intraoperative hypotension and fluid resuscitation, within-patient changes in PVP mirrored changes in CVP (r = 0.92). CONCLUSION: In children undergoing major surgery, PVP showed good agreement with CVP in the perioperative period. As changes in PVP parallel, in direction, changes in CVP, PVP monitoring may offer an alternative to direct CVP measurement for perioperative estimation of volume status and guiding fluid therapy. PMID- 15352956 TI - Clinical application of the pO(2)-pCO(2) diagram. AB - Based on the classic, linear blood gas diagram a logarithmic blood gas map was constructed. The scales were extended by the use of logarithmic axes in order to allow for high patient values. Patients with lung disorders often have high arterial carbon dioxide tensions, and patients on supplementary oxygen typically respond with high oxygen tensions off the scale of the classic diagram. Two case histories illustrate the clinical application of the logarithmic blood gas map. Variables from the two patients were measured by the use of blood gas analysis equipment. Measured and calculated values are tabulated. The calculations were performed using the oxygen status algorithm. When interpreting the graph for a given patient it is recommended first to observe the location of the marker for the partial pressure of oxygen in inspired, humidified air (I) to see whether the patient is breathing atmospheric air or air with supplementary oxygen. Then observe the location of the arterial point (a) to see whether hypoxemia or hypercapnia appears to be the primary disturbance. Finally observe the alveolo arterial oxygen tension difference to estimate the degree of veno-arterial shunting. If the mixed venous point (v) is available, then observe the value of the mixed venous oxygen tension. This is the most important indicator of global tissue hypoxia. PMID- 15352957 TI - Cardioprotective action of fentanyl in a model of central sympathetic overactivity in rabbits: antiarrhythmic and anti-ischemic effects. AB - BACKGROUND: Sympathetic overactivity resulting from perioperative noxious stimuli elicits hyperdynamic cardiovascular responses that may lead to myocardial ischemia and/or ventricular arrhythmia, especially in patients presenting with coronary artery disease. In the present study we investigated the cardioprotective effects of clinically relevant doses of fentanyl in an experimental model of sympathetic overactivity associated with myocardial ischemia in anesthetized rabbits. METHODS: Central sympathetic stimulation was achieved through intracerebroventricular (i.c.v.) injection of L-glutamate (10 micro mol), with simultaneous inhibition of nitric oxide (NO) synthesis through i.v. administration of N(omega)-nitro-l-arginine methyl ester (L-NAME; 40 mg kg( 1)). RESULTS: L-glutamate triggered ventricular arrhythmia and electrocardiographic alterations indicative of myocardial ischemia. The intravenous administration of fentanyl (5, 10 or 50 micro g kg(-1)) reduced the incidence of ST-segment shift (70, 20 and 10%, respectively, vs. 66.7% in controls) as well as of T-wave inversion from 58.3% to 30, 20 and 10%, respectively. The total number of ventricular premature complexes per minute fell from 65.2 +/- 16 in the control group to 6.8 +/- 3, 3.5 +/- 2 and 2.6 +/- 1.5, respectively. The occurrence of ventricular tachycardia and bigeminy was completely abolished by fentanyl. Finally, the i.v. administration of fentanyl did not induce significant hemodynamic effects (except for dP/dt(max) in the 50 micro g kg(-1)-dose). CONCLUSION: Fentanyl elicits significant cardioprotective effects in a model of arrhythmia resulting from the association of central sympathetic overactivity with myocardial ischemia in rabbits, independently from its systemic hemodynamic actions. PMID- 15352958 TI - Oleic acid lung injury: a morphometric analysis using computed tomography. AB - BACKGROUND: The oleic acid-induced lung injury (OAI) model is considered to represent the early phase of acute respiratory distress syndrome (ARDS). Its inherent properties are important for the design and the interpretation of interventional studies. The aim of this study was to describe the evolution of morphometric lung changes during OAI using computed tomography (CT) analysis. Furthermore, the effect of a temporary change in positive end-expiratory pressure (PEEP) was evaluated. METHODS: Fifteen anaesthetized pigs were ventilated in volume-controlled mode with a baseline PEEP of 5 cm H(2)O. Helical CT scans were taken at baseline and 1 h after oleic acid injection. The PEEP was then either increased to 10 cm H(2)O (n = 5), decreased to 0 cm H(2)O (n = 5) or kept constant (n = 5) for 30 min. For the next 30 min, the baseline PEEP level was applied in all animals before the final CT scans 2 h after the induction of OAI. Dimensional and volumetric changes were determined from radiographical attenuation values. RESULTS: There was a major decrease in gas volume and an increase in tissue volume within the first hour. A net increase in total lung volume, with a larger transverse area but no displacement of the diaphragm, was manifest after 2 h. A minor increase in volume of non-aerated lung, located to the caudal region, was observed during the second hour. The tidal volume was redistributed to the middle and apical regions. The temporary change in PEEP did not influence the morphological progress of OAI. CONCLUSION: Decreased gas volume and increased tissue volume are the dominating morphometric characteristics of oleic acid lung injury, occurring mainly within the first hour. With these changes manifest, the course of injury is not affected by a limited period of moderately changed PEEP during the second hour. The net increase of total lung volume suggests a predominance of oedema formation over airway and alveolar collapse. PMID- 15352959 TI - 'Protective premedication': an option with gabapentin and related drugs? A review of gabapentin and pregabalin in in the treatment of post-operative pain. AB - Substantial progress has been made during the last decades in our understanding of acute pain mechanisms, and this knowledge has encouraged the search for novel treatments. Of particular interest has been the observation that tissue injury initiates a number of modulations of both the peripheral and the central pain pathways, which convert the system from a 'physiological' to a 'pathological' mode of processing afferent information. Gabapentin, which binds to the alpha(2)delta subunit of the voltage-dependent calcium channel, is active in animal models of 'pathological' but not in models of 'physiological' pain. Consequently, attention has so far been focused on neuropathic pain as a target for the clinical use of gabapentin and analogues. Recently, several reports have indicated that gabapentin may have a place in the treatment of post-operative pain. This article presents a brief summary of the potential mechanisms of post operative pain, and a systematic review of the available data of gabapentin and pregabalin for post-operative analgesia. It is concluded that the results with gabapentin and pregabalin in post-operative pain treatment published so far are promising. It is suggested that future studies should explore the effects of 'protective premedication' with combinations of various antihyperanalgesic and analgesic drugs for post-operative analgesia. PMID- 15352960 TI - Postoperative cognitive dysfunction: true deterioration versus random variation. AB - BACKGROUND: Postoperative cognitive dysfunction (POCD) is a common complication, especially in the elderly. The aim of this study was to describe how variability in neuropsychological testing could lead to the detection of cognitive improvement and poor consistency of POCD between postoperative test sessions. METHODS: In four published studies performed by the ISPOCD group, we included 2536 patients and 359 healthy controls. Cognitive function was assessed using neuropsychological tests preoperatively and at 7 days and 3 months thereafter, comparing the changes between those at baseline with those after surgery. Postoperative cognitive dysfunction was defined as a Z score greater than 2, and we also defined a corresponding improvement as a Z score less than -2. Consistency of POCD between postoperative test sessions was analyzed and we also assessed test-retest variability using data from healthy control subjects. RESULTS: Improvement in cognitive function was found in 4.2-8.7% of patients after 1 week and in 5.0-7.8% after 3 months. The ratio between incidence of dysfunction and improvement varied in patients between 3.3 and 6.2 early after major surgery. Of those patients who displayed POCD at the 3-month test, 30-48% also had POCD at the previous test at 1 week. The test-retest reliability was between 0.56 and 0.90, except for the error score in Concept Shifting Test, where the values were 0.20 and 0.37. CONCLUSION: Variability in neuropsychological test data contributes to a low consistency between postoperative test sessions but it does not explain the detection of cognitive dysfunction after major surgery. PMID- 15352962 TI - Supplemental remifentanil during coronary artery bypass grafting is followed by a transient postoperative cardiac depression. AB - BACKGROUND: The pharmacokinetic properties of the short-acting micro opioid receptor-agonist remifentanil makes it possible to give cardiac surgical patients a deep intraoperative anesthesia without experiencing postoperative respiratory depression and a prolonged stay in the intensive care unit (ICU). However, previous investigations have shown that patients who received remifentanil required additional analgesia during the early postoperative period as compared to patients who received fentanyl. The aim of the present study therefore was to investigate the effects of supplementing remifentanil to a standard fentanyl based anesthesia in coronary artery bypass grafting (CABG). METHODS: The study was prospective, randomized, double-blind, and placebo-controlled. Twenty male patients aged 55-70 years were included. All patients received a standard fentanyl and isoflurane-based anesthesia. In addition, the patients were randomized to receive either remifentanil 0.5 micro g kg(-1) min(-1) or placebo during surgery. Hemodynamic recordings and measurements of blood glucose and plasma adrenaline and noradrenaline were performed intra- and postoperatively. RESULTS: Remifentanil reduced the hemodynamic and metabolic response to surgical stress compared to the standard fentanyl-based anesthetic regimen. However, the patients in the remifentanil group had a lower cardiac output (CO), left ventricular stroke work index (LVSWI), and mixed venous oxygen saturation (SvO(2)), and a higher central venous pressure (CVP) than the patients in the placebo group during the early postoperative phase, indicating a postoperative cardiac depression in the remifentanil group. CONCLUSION: In CABG, remifentanil reduces the hemodynamic and metabolic responses during surgery but seems to give a cardiac depression in the early postoperative phase. PMID- 15352961 TI - An anaesthetic protocol in the young domestic pig allowing neuromuscular blockade for studies of cardiac function following cardioplegic arrest and cardiopulmonary bypass. AB - BACKGROUND: Neuromuscular blockade should, for ethical reasons, not be allowed in animal experiments unless the use is strongly motivated. Beforehand, the anaesthetic protocol must be documented without muscle relaxation in the species studied. Documentation is difficult to obtain from the scientific literature. When focusing on cardiac function over time, in particular, the ideal anaesthetic protocol should cause no or minor alterations in cardiac variables. METHODS: We intended to document an anaesthetic protocol involving ventilation with N(2)O combined with loading doses and continuous infusions of pentobarbital, fentanyl and midazolam in seven pigs by applying potentially painful stimuli every 15 min for 7 h. Subsequently, left ventricular global and regional function was studied with conductance catheter and strain rate imaging by echocardiography in eight pigs with pancuronium included. RESULTS: Pigs without pancuronium were completely immobilized and unresponsive to potentially painful stimuli and sternotomy, with no accumulation or degradation of anaesthetic agents. With pancuronium included, left ventricular preload gradually decreased together with reduction of cardiac index from 3.52 +/- 0.14 at 2 h to 2.84 +/- 0.11 L min(-1). m(-2) (+/-SEM) after 7 h of observation. Preload recruitable stroke work decreased after 7 h, whereas peak systolic strain in the anterior left ventricular wall and load-independent indices of diastolic function were not significantly altered. CONCLUSION: In specific experimental protocols, the anaesthetic protocol described could allow the use of muscular paralysis in young domestic pigs, for instance when involving hypothermic cardiopulmonary bypass, cardioplegic arrest and reperfusion. PMID- 15352963 TI - Effects of nitrous oxide on baroreflex gain and heart rate variability. AB - BACKGROUND: Spontaneous baroreflex method allows continuous assessment of cardiovagal reflex function within resting blood pressure, but effects of nitrous oxide, per se, on the spontaneous baroreflex response remain unknown. This study was designed to determine the effects of nitrous oxide on spontaneous baroreflex gain and heart rate variability assessed by power spectral analysis in humans. METHODS: Electrocardiogram and non-invasive blood pressure were monitored in 12 healthy volunteers before and during a 15-min inhalation of 67% nitrous oxide in oxygen, while spontaneous respiration was maintained. Least-square regression analysis relating R-R interval and systolic blood pressure was performed to obtain spontaneous baroreflex gains. Heart rate variability was analyzed using fast Fourier transformation. RESULTS: Nitrous oxide did not significantly alter spontaneous baroreflex gains, which correlated well with high-frequency power (0.15-0.4 Hz) of heart rate variability before and during nitrous oxide inhalation. CONCLUSION: Our results indicate that (a) cardiovagal reflex response is not affected by nitrous oxide, per se, and (b) spontaneous baroreflex responses closely reflect beat-to-beat dynamic modulation of the cardiac cycle by the parasympathetic nervous system during inhalation of 67% nitrous oxide. PMID- 15352964 TI - Comparison of BIS and AAI as measures of anaesthetic drug effect during desflurane-remifentanil anaesthesia. AB - BACKGROUND: Autoregressive modelling with exogenous input of the middle-latency auditory evoked potential has been developed for monitoring of anaesthetic depth. This study was designed to investigate the dose-response relationship between endtidal desflurane concentrations and the Alaris Autoregressive Index (AAI, Alaris Medical, Hampshire, UK, version 1.4) or the bispectral index (Aspect Medical Systems, Newton, MA, USA, version XP). METHODS: Twenty-one patients scheduled for radical prostatectomy were investigated. After premedication and induction of anaesthesia with propofol und remifentanil all patients received atracurium and a remifentanil background infusion at a constant rate of 0.1 micro g kg(-1) min(-1). During dissection of the prostate, desflurane endtidal concentrations were varied between 3 and 9 vol%. Both AAI and BIS were determined and compared with the respective endtidal desflurane concentration. RESULTS: None of the patients showed a significant change of AAI values while changing the desflurane concentrations between 3 and 9 vol%. The dose-response of BIS values and desflurane concentrations was not uniform: two patients showed increasing BIS values with increasing desflurane concentrations, while in three patients BIS values remained unchanged. In 16 patients decreasing BIS values adequately reflected an increase in desflurane concentrations. CONCLUSION: Changes of desflurane concentrations during deep anaesthesia were adequately displayed only in 16 of 21 cases by BIS but in none of the cases by AAI monitoring. PMID- 15352965 TI - Electromyographic assessment of blink reflexes correlates with a clinical scale of depth of sedation/anaesthesia and BIS during propofol administration. AB - BACKGROUND: General anaesthesia is characterized by loss of consciousness, amnesia and obtundation of reflex responses to noxious stimuli. Quantifying the blink reflex may reflect the depression of reflex arches induced by anaesthetics and thus being informative on the anaesthetic state. METHODS: The relation between the electrically evoked blink reflexes and the depth of sedation and anaesthesia induced with intravenous propofol was investigated. Twenty patients received propofol by target-controlled infusion to create a stepwise deepening of sedation and anaesthesia. Depth of anaesthesia was assessed using the observer's assessment of anaesthesia and sedation (OAAS) scale, and by bispectral EEG analysis (BIS). Probit analysis was used to estimate the predicted propofol effect site concentrations producing unconsciousness, no response to noxious stimulation, and loss of blink reflex components. RESULTS: Latency of the first (R1) and second (R2) blink component increased, whereas duration and area decreased with increasing depth of sedation and anaesthesia. A reasonably strong correlation between OAAS and the areas of R1 and R2 components was found (Spearman's rho = 0.92 and 0.89). The areas of R1 and R2 and the OAAS also correlated with BIS (Spearman's rho = 0.91, 0.88 and 0.90). EC(50) and EC(95) for loss of R1 were 2.8 (95% CI: 2.5-3.2) micro g/ml and 4.6 (95% CI: 4.1-5.5) micro g/ml, respectively. CONCLUSIONS: Our results suggest that the differential sensitivity of the components of the blink reflex could be useful in monitoring depth of sedation and light levels of anaesthesia during the administration of propofol. Both OAAS and BIS correlate similarly with the blink reflex components. PMID- 15352966 TI - Nitrous oxide diffusion into tracheal tube cuffs: comparison of five different tracheal tube cuffs. AB - BACKGROUND: The aim of this study was to investigate cuff compliance and cuff pressure during nitrous oxide exposure in the recently introduced Microcuff tracheal tube with a polyurethane cuff (Microcuff GmbH, Weinheim, Germany), and to compare it to conventional tracheal tubes with PVC cuffs. METHODS: In an in vitro set up, five cuffed tracheal tubes (ID 7.0 mm) from different manufacturers (Microcuff HVLP, Portex Profile Soft Seal, Mallinckrodt HiLo, Rusch Super Safety Clear and Sheridan CF) were studied. Pressure-volume curves were assessed and changes of cuff pressure during exposure to nitrous oxide (for 60 min; 66% N(2)O in oxygen) were recorded without and with restriction of the cuff in a trachea model. Each experiment was performed four times using two exemplars of each tube twice. Sixty-minute values of the Microcuff group were compared with the other groups using the Mann- Whitney test (P < 0.05). RESULTS: The Microcuff polyurethane cuff demonstrated an intermediate cuff compliance but the highest cuff pressure increase during nitrous oxide exposure under unrestricted conditions. When inflated within the artificial trachea its cuff compliance became the highest of all tested tracheal tubes. However, exposure to N(2)O, again led to a rapid increase in cuff pressure. CONCLUSION: The ultra-thin polyurethane tube cuff demonstrated higher permeability for nitrous oxide than conventional PVC cuffs and led to a rapid cuff pressure increase when exposed to N(2)O. Routinely checking of cuff-pressure or filling the cuff with nitrous oxide are more important than the brand of tube used. PMID- 15352967 TI - Propacetamol and diclofenac alone and in combination for analgesia after elective tonsillectomy. AB - BACKGROUND: Diclofenac and paracetamol have different mechanisms and sites of action. Therefore, we tested if their combination is more effective for analgesia after tonsillectomy than either drug alone with respect to rescue analgesic consumption and visual analog scale values. METHODS: The analgesic effects of intravenously administered propacetamol (injectable pro-drug of paracetamol) and diclofenac or a combination on postoperative pain were compared in 71 adult elective tonsillectomy patients in a randomized, double-blind study. After induction of anesthesia the patients received monotherapy with 2 g propacetamol (n = 25) or 75 mg diclofenac (n = 25), or a combined treatment with 2 g propacetamol and 75 mg diclofenac (n = 21) in physiologic saline as an infusion. Postoperatively the propacetamol dosage was repeated twice and diclofenac once on the ward. Oxycodone (0.03 mg kg(-1)) was used as a rescue analgesic by patient controlled analgesia. RESULTS: On average the patients needed oxycodone 15.3, 13.2 and 10.6 times in the propacetamol, diclofenac and combination groups, respectively (NS). A verbal rating scale and a visual analog scale were employed for assessing post-tonsillectomy pain, nausea and patient satisfaction in all groups. No statistically significant differences were found between the groups. Twelve of the 25 (48%) patients having received propacetamol complained of pain at the cannulation site. CONCLUSION: Combined treatment with propacetamol and diclofenac with the dosages used provided clinically only a minor advantage over monotherapy with propacetamol or diclofenac with respect to postoperative analgesia or the incidence of side-effects in adult tonsillectomy patients. PMID- 15352968 TI - Efficacy of oral rofecoxib versus intravenous ketoprofen as an adjuvant to PCA morphine after urologic surgery. AB - BACKGROUND: Adjunctive use of nonsteroidal anti-inflammatory drugs has become increasingly popular in the perioperative period because of their opioid-sparing effects. This randomized, controlled, double-dummy study was designed to evaluate the cost-effectiveness of using oral rofecoxib as an alternative to intravenous ketoprofen for pain management in patients undergoing urologic surgery. METHODS: Seventy patients were randomly assigned to receive either a placebo (Control) or rofecoxib 50 mg po (Rofecoxib) 1 h prior to surgery. After a standardized spinal anesthetic, patients in the Control group received ketoprofen 100 mg IV q 8 h for 24 h, while the Rofecoxib group received an equivolume of saline at 8-h intervals for 24 h. Both groups were allowed to self-administer morphine (1 mg IV boluses) using a PCA delivery system. The need for 'rescue' analgesic medication, as well as pain scores [using an 11-point verbal rating scale (VRS) (0 = none to 10 severe)], were recorded at 1, 2, 6, 12, and 24-h intervals after surgery. In addition, the incidences of side-effects were recorded at the end of the study period. RESULTS: Total amount of morphine required in the initial 24-h postoperative period was nonsignificantly reduced in the Rofecoxib group (29 +/- 2 vs. 37 +/- 4 mg). More importantly, the percentage of patients reporting moderate-to-severe pain (VRS score > or =4) during the study period was lower in the Rofecoxib group (12 vs. 22%, P < 0.05). The daily cost of rofecoxib (USD 1.14 for 50-mg dose) was also significantly less than ketoprofen (USD 3.06 for three 100-mg doses). CONCLUSION: Premedication with oral rofecoxib (50 mg) is a cost effective alternative to the parenteral nonselective NSAID, ketoprofen (100 mg q 8 h), when used as an adjuvant to PCA morphine for pain management after urologic surgery. PMID- 15352969 TI - Presurgical intravenous parecoxib sodium and follow-up oral valdecoxib for pain management after laparoscopic cholecystectomy surgery reduces opioid requirements and opioid-related adverse effects. AB - BACKGROUND: Opioids are associated with numerous adverse effects. It is unclear if reduced postoperative opioid consumption lowers the incidence and severity of opioid-related adverse effects. This analysis -- from a multicenter, randomized, double-blind trial -- tested if the reduction of opioid consumption among patients who received intravenous preoperative parecoxib 40 mg, followed by oral valdecoxib 40 mg qd postoperatively, in Days 1-4 after outpatient laparoscopic cholecystectomy surgery, reduced opioid-related symptoms. METHODS: Patients received intravenous fentanyl for pain before discharge, and oral acetaminophen 500 mg hydrocodone 5 mg q 4-6 h prn postdischarge for up to 7 days postsurgery. Patients also received intravenous parecoxib 40 mg administered 30-45 min preoperatively, and valdecoxib 40 mg qd up to Day 4 and prn Days 5-7 postsurgery, or placebo. Patients completed an opioid-related Symptoms Distress Scale (SDS) questionnaire every 24 h for 7 days. Opioid use was converted to morphine equivalent doses (MEDs). Clinically meaningful events (CMEs) for 12 opioid related symptoms were assessed by three ordinal measures: frequency, severity, and bothersomeness. Reduction of CMEs on Day 1 and number of patient-days with CMEs on Days 1-4 were examined. RESULTS: Cumulative MEDs on Day 0, Day 1, and Days 1-4 were significantly lower in the parecoxib/valdecoxib group compared with the placebo group (P < 0.001). At the end of Day 1, parecoxib/valdecoxib-treated patients had significantly lower SDS scores (P < 0.02), a significantly reduced incidence of CMEs (P < 0.05), and significantly fewer patient-days with CMEs in Days 1-4 than placebo patients (P < 0.05). Patients in the parecoxib/valdecoxib group were less likely to have CMEs for multiple symptoms than those in the placebo group (P < 0.001). CONCLUSIONS: Treatment with parecoxib and valdecoxib significantly reduced the cumulative MED requirements, the incidence of opioid related adverse effects, and patient-days with CMEs. PMID- 15352970 TI - Gas embolism: an exceptional complication of radial arterial catheterization. AB - BACKGROUND: We report a rare complication of radial arterial catheterization in a 74-year-old man who had undergone retroperitoneal surgery for an infra-renal aortic aneurysm. A right subclavian venous catheter and a right radial artery catheter were inserted for hemodynamic monitoring. RESULTS: The patient suddenly went into a coma, with dyspnea and bradycardia, 1 day postsurgery, as a result of a cerebral gas embolism produced by the accidental entry of pressurized air into the artery via the arterial catheter. Cerebral Magnetic Resonance Imaging revealed multiple, diffuse brain lesions. Six months later the patient still had a left hemiparesis and a cognitive deficit. He walks with assistance. CONCLUSION: Misuse of an arterial catheter can lead to a severe gas embolism. The infusion system used to flush arterial catheters should be checked regularly to ensure it contains no gas. PMID- 15352971 TI - Total spinal anaesthesia as a complication of local anaesthetic test-dose administration through an epidural catheter. AB - We describe a case of total spinal anaesthesia, which occurred after a 3-ml lignocaine (20 mg ml(-1)) test dose was administered through an epidural catheter in a 79-year-old patient scheduled for gastrectomy under combined general and epidural anaesthesia. The surgery was postponed, and the patient required admission to the intensive therapy unit. Spinal MRI from the total spinal cord did not reveal any pathology. During the next 24 h the patient recovered and after 11 days was successfully operated on under general anaesthesia. No late complications followed. We presume that during placement, the epidural catheter had migrated to the spinal canal as a result of technical difficulties. Although controversial, we consider that administering a standard test dose of local anaesthetic via an epidural catheter is recommended, especially in high-risk patients and when epidural space identification or catheter placement poses technical difficulties. A test dose of local anaesthetic does not fully prevent complications. PMID- 15352972 TI - Fetal bradycardia and disseminated coagulopathy: atypical presentation of amniotic fluid emboli. AB - We present a parturient who developed an atypical case of amniotic fluid emboli presented by sudden fetal bradycardia, followed by maternal disseminated coagulopathy. The typical feature of cardiopulmonary collapse was absent in this patient implying that in some cases of amniotic fluid emboli (AFE), fetal hypoxia or acidemia is unrelated to maternal cardiopulmonary status. PMID- 15352973 TI - Sevoflurane and seizures: deja vu. PMID- 15352974 TI - Bilateral occipital neuropathy, not so rare. PMID- 15352975 TI - Malaria and postoperative fever. PMID- 15352976 TI - Epidural anesthesia for laminectomy in a geriatric patient. PMID- 15352977 TI - Reactive versus neoplastic lymphoid follicles: proliferation and death versus quiescence and staying alive. PMID- 15352978 TI - Post-transplant air-leak syndrome. PMID- 15352979 TI - Genetic basis of blood group diversity. AB - In the last 18 years the genes that encode all but one of the 29 blood group systems present on red blood cells (RBCs) have been identified. This body of knowledge has permitted the application of molecular techniques to characterize the common blood group antigens and to elucidate the background for some of the variant phenotypes. Just as the RBC was used as a model for the biochemical characterization of cell membranes, so the genes encoding blood groups provide a readily accessible model for the study of gene expression and diversity. The application of genotyping techniques to identify fetuses at risk of haemolytic disease of the newborn is now the standard of care, and the expansion of nucleic acid testing platforms to include both disease testing and blood typing in the blood centre is on the horizon. This review summarizes the molecular basis of blood groups and illustrates the mechanisms that generate diversity through specific examples. PMID- 15352980 TI - The role of PET imaging in lymphoma. AB - Positron emission tomography using fluorine-18 (FDG-PET) is increasingly used in the staging and follow-up of malignant lymphomas, although its precise role has not yet been determined. This review considers the results reported at the different stages in the disease history and separately considers the major histological subtypes. Attention is given to the situations in which PET scanning is most likely to influence management. Finally, this review discusses ongoing developments in PET scanning with improved resolution and different radiolabelled tracers. PMID- 15352981 TI - FLT3 mutations in myeloid sarcoma. AB - Myeloid sarcoma is an extramedullary tumour that typically occurs in the setting of acute myeloid leukaemia (AML), or myeloproliferative disorders. In AML, two types of mutations in Fms-like tyrosine kinase 3 (FLT3) have been described; internal tandem duplications (ITD) and point mutations at aspartic acid residue 835 (D835). We analysed 24 myeloid sarcoma specimens from 20 patients for FLT3 ITD and D835 mutations. FLT3 ITD mutations were identified in three of 20 cases (15%); no D835 mutations were identified. The ITD inserts ranged in size from 33 to 198 base pairs (bp) and represented approximately 20-40% of the FLT3 alleles. Two cases showed discordance in FLT3 ITD mutational status. In one case, the leukaemia specimen was positive for a FLT3 ITD mutation and the myeloid sarcoma specimen was negative. In the second case, the myeloid sarcoma was positive for a FLT3 ITD mutation at diagnosis, but negative in subsequent relapse samples. Our findings suggest that small molecule inhibitors of FLT3 may be useful therapeutic agents for treatment of myeloid sarcomas-containing FLT3 mutations, however, the potential for discordance between the leukaemia and myeloid sarcoma, necessitates that the myeloid sarcoma tumour itself be analysed for FLT3 mutations. PMID- 15352982 TI - A randomized study (WOS MM1) comparing the oral regime Z-Dex (idarubicin and dexamethasone) with vincristine, adriamycin and dexamethasone as induction therapy for newly diagnosed patients with multiple myeloma. AB - Whilst infusional vincristine, adriamycin and dexamethasone (VAD) is an effective treatment for patients with multiple myeloma (MM), administration may be complicated by line-associated infections and thromboses. The oral regime, Z-Dex (idarubicin and dexamethasone) has been shown to be efficacious in MM. We conducted a randomized study comparing Z-Dex with VAD as induction therapy in newly diagnosed MM patients. A total of 106 patients (median age, 56 years; range: 37-73; Durie-Salmon stage II/III) were randomized to receive four to six cycles of Z-Dex or VAD. Central line complications were reported in 38 patients on 57 cycles, primarily because of infection. Neutropenia (all grades) was more common in the Z-Dex arm (P = 0.009) although grade III/IV neutropenia was not significantly different between the treatment groups (P = 0.06). Infections (all grades) were more commonly seen in the VAD arm (P = 0.001) although grade III/IV infections were not significantly different between the two groups (P = 0.081). The responses to therapy (complete/partial response) in evaluable patients were: VAD 74% vs. Z-Dex 58%, with an estimated difference in response of 16% (95% CI -2 33, P = 0.075). VAD recipients (15%) suffered early treatment-related mortality compared with 12% of Z-Dex recipients. Overall, 45 patients have died: disease progression (Z-Dex n = 13, VAD n = 10), regimen-related toxicity (Z-Dex n = 2, VAD n = 2), infection (Z-Dex n = 0, VAD n = 3), other causes (Z-Dex n = 7, VAD n = 2), unknown (Z-Dex n = 3, VAD n = 2). This study demonstrated that Z-Dex might be a suitable oral alternative to VAD for treating newly diagnosed MM patients, although definitive evidence for equivalence is not provided. PMID- 15352983 TI - Treating childhood acute myeloid leukaemia with the AML-BFM-83 protocol: experience in a developing country. AB - Treatment for childhood acute myeloid leukaemia (AML) consists of remission induction chemotherapy followed by postremission chemotherapy with or without bone marrow transplantation. The AML Berlin-Frankfurt-Munster (BFM)-83 protocol with induction-consolidation-maintenance chemotherapy for 2 years has been reported to result in a 6-year event-free survival (EFS) and event-free interval (EFI) of 49% and 61% respectively. A total of 174 Malaysian children were treated with this protocol between 1985 and 1999. The 5-year EFS and EFI was 30.7% and 48.0% respectively. The overall mortality from sepsis was 24%, which needs urgent address. The 5-year EFS for patients treated before 1993 and after 1993 was 18.6% and 41.3%, respectively (P = 0.04), while the EFI was 32% and 60.6% respectively (P = 0.034). The improvement seen after 1993 was related to a reduction in induction deaths for that period and probably reflected increased capability and familiarity to cope with the demands of the AML-BFM-83 protocol and accompanying complications in the treatment of AML. PMID- 15352984 TI - Impaired generation of bone marrow CD34-derived dendritic cells with low peripheral blood subsets in patients with myelodysplastic syndrome. AB - Myelodysplastic syndrome (MDS) is a stem cell disorder characterized by ineffective haematopoiesis and blood cytopenias. The present study investigated the potential of bone marrow CD34(+) progenitors in MDS patients to proliferate and differentiate into dendritic cells (DCs) in a cytokine-supplemented liquid culture system and analysed the status of blood DC subsets in these patients. CD34(+) progenitors had low potential to generate DCs in vitro, as the number of DCs obtained from one CD34(+) cell was significantly lower compared with controls (median value 0.2 vs. 4, P = 0.003). In patients, the survival and proliferation of CD34(+) cells in culture was not correlated to the degree of apoptosis. Phenotypically and functionally CD34(+)-derived DCs were similar in MDS patients and normal subjects. The percentage of both circulating DC subsets in patients was extremely diminished compared with controls (myeloid DC: 0.10 +/- 0.10% vs. 0.35 +/- 0.13%, P < 0.001; plasmacytoid DC: 0.11 +/- 0.10% vs. 0.37 +/- 0.14%, P < 0.001). In cases with the 5q deletion both CD34-derived DCs and blood DCs harboured the cytogenetic abnormality. Our results indicate that, in MDS, the production of DCs is affected by the neoplastic process resulting in ineffective 'dendritopoiesis' with low blood DC precursor numbers. This quantitative DC defect probably contributes to the poor immune response against infectious agents and to the escape of the malignant clone from immune recognition with disease progression towards acute leukaemia. PMID- 15352985 TI - High response rate and manageable toxicity with an intensive, short-term chemotherapy programme for Burkitt's lymphoma in adults. AB - A very short, intensive paediatric chemotherapy programme was tested in a consecutive monoinstitutional group of 22 adult Burkitt's lymphoma (BL) patients. After a 5-week induction phase of weekly infusions consisting of vincristine, cyclophosphamide, doxorubicin, high-dose (HD) methotrexate (MTX) plus leukovorin rescue, and intrathecal MTX or cytarabine (ARA-C), a consolidation phase including HD ARA-C plus cisplatin was given. Responding patients achieving less than complete response (CR) after completion of the initial induction phase, were promptly shifted to a high-dose, stem cell supported sequential chemotherapy schema (R-HDS). PATIENT CHARACTERISTICS: median age, 35.5 (range 18-76) years; Ann Arbor stage I-II/III-IV, 11/11; bulky disease, 15 patients; LDH > or = 460 U/l, 11 patients. The median duration of the chemotherapy programme was 62 d (range, 43-94 d). Seventeen patients achieved a CR (77%), one patient died of progressive disease and four partial responders following induction were converted to CR following R-HDS. Of 17 patients in CR, one died of infectious toxicity while in CR, and one relapsed at 30 months and died of progressive disease. After a median follow-up of 28.7 months (range, 6-158 months), 16 patients (73%) were in continued CR. Overall survival and progression-free survival were 77% [95% confidence interval (CI), 52-99%] and 68% (95% CI, 43-99%) respectively. Confirmation of these excellent efficacy and feasibility results by larger, multicentre and prospective studies is warranted. PMID- 15352986 TI - Dual targeting of Bruton's tyrosine kinase and Janus kinase 3 with rationally designed inhibitors prevents graft-versus-host disease (GVHD) in a murine allogeneic bone marrow transplantation model. AB - The purpose of the present study was to evaluate the effectiveness of targeting Bruton's tyrosine kinase (BTK) with a specific BTK inhibitor, alpha-cyano-beta hydroxy-beta-methyl-N-(2,5-dibromophenyl)-propenamide (LFM-A13), for prevention of acute fatal graft-versus-host disease (GVHD) in a murine model of allogeneic bone marrow transplantation (BMT). Vehicle-treated control C57BL/6 mice receiving bone marrow/splenocyte grafts from allogeneic BALB/c donors developed severe multi-organ acute GVHD and died after a median survival time (MST) of 40 d. LFM A13 treatment (25 mg/kg/d) significantly prolonged the MST of the BMT recipients to 47 d. The probability of survival at 2 months after BMT was 2 +/- 2% for vehicle-treated control mice and 22 +/- 6% for mice treated with LFM-A13 (P = 0.0008). Notably, the combination regimen of LFM-A13 plus the standard anti-GVHD drug methotrexate (MTX) (10 mg/m(2)/d) was more effective than LFM-A13 alone, while the combination regimen of LFM-A13 plus the novel anti-GVHD drug JANEX-1 (60 mg/kg/d), targeting Janus kinase 3, was more effective than LFM-A13, JANEX-1 or MTX alone. More than 70% of recipients receiving this most effective GVHD prophylaxis (LFM-A13 + JANEX-1) remained alive throughout the 80-d observation period with an MST of >80 d. Taken together, these results indicate that targeting BTK with the chemical inhibitor LFM-A13 may attenuate the severity of GVHD, especially when it is combined with other anti-GVHD drugs, such as MTX and JANEX-1. PMID- 15352987 TI - Induction of acute graft-versus-host disease by T cells that do not respond to in vitro alloantigen stimulation. AB - The mixed lymphocyte reaction (MLR) has been used extensively to measure alloreactive T cells. In clinical practice, a negative MLR of recipient T cells responding to donor cells does not necessarily mean that a donor-specific tolerance has been established. This discrepancy indicates that the presently used methods fail to demonstrate the full repertoire of alloreactive T cells. This could be the result of the fact that some alloreactive T cells do not respond in vitro but will mount a response towards alloantigens in vivo, or that some alloreactive T cells do not respond during the MLR but will respond later if the alloantigen stimulation remains. We therefore examined the non-proliferating T-cell population in a mouse primary alloreactive response. Spleen and lymph node cells derived from C57BL/6J (H-2(b)) mice were stained with carboxy-fluorescein diacetate succinimidyl ester and injected intravenously into C.B-17 severe combined immunodeficient (SCID) mice (H-2(d)). The donor cells were recovered 5 d after the injection. The non-proliferating T cells were sorted and were non reactive to alloantigen stimulation in vitro. Nevertheless, these non proliferating T cells could proliferate and cause acute graft-versus-host disease after being adoptively transferred to secondary recipients of SCID mice. These results suggest that there exists an alloreactive T-cell population that does not respond to in vitro alloantigen stimulation but can mount a delayed alloresponse in vivo. PMID- 15352988 TI - Targeted pretransplant host lymphocyte depletion prior to T-cell depleted reduced intensity allogeneic stem cell transplantation. AB - Mixed chimaerism and graft rejection are higher after reduced-intensity allogeneic stem cell transplantation (RIST) with T-cell depleted (TCD) allografts. As host immune status before RIST affects engraftment, we hypothesized that targeted depletion of host lymphocytes prior to RIST would abrogate graft rejection and promote donor chimaerism. Lymphocyte-depleting chemotherapy was administered at conventional doses to subjects prior to RIST with the intent of decreasing CD4(+) counts to <0.05 x 10(9)cells/l. Subjects (n = 18) then received reduced-intensity conditioning followed by ex vivo TCD human leucocyte antigen-matched sibling allografts. All evaluable patients (n = 17) were engrafted; there were no late graft failures. At day +28 post-RIST, 12 patients showed complete donor chimaerism. Mixed chimaerism in the remaining five patients was associated with higher numbers of circulating host CD3(+) cells (P = 0.0032) after lymphocyte-depleting chemotherapy and was preferentially observed in T lymphoid rather than myeloid cells. Full donor chimaerism was achieved in all patients after planned donor lymphocyte infusions. These data reflect the importance of host immune status prior to RIST and suggest that targeted host lymphocyte depletion facilitates the engraftment of TCD allografts. Targeted lymphocyte depletion may permit an individualized approach to conditioning based on host immune status prior to RIST. PMID- 15352989 TI - IL-7 effect on immunological reconstitution after HSCT depends on MHC incompatibility. AB - Considerable progress has been recently accomplished in the management of patients who have undergone haplo-incompatible haematopoietic stem cell transplantation (HSCT) in terms of intake and prevention of graft-versus-host disease. Nevertheless haplo-incompatible HSCT is a procedure limited to a small number of patients because of the long-lasting immunodeficiency that is responsible for more than 50% of deaths within the first 3 months. Interleukin (IL)-7, which plays a unique and key role in T-cell development both in the mouse and in the human, is particularly attractive for attempting to speed up T-cell reconstitution. However, controversial results have been obtained after bone marrow graft in murine and primate models. To elucidate the impact of IL-7 treatment, we have performed HSCT in irradiated murine recombination activating gene (RAG) immunodeficient recipients, using donors that exhibited increased major histocompatibilty complex (MHC) incompatibility. Although irradiation performed prior to HSCT lead to a profound defect in the thymic stromal cells responsible for IL-7 production, IL-7 treatment had no significant effect on immune reconstitution in the MHC compatible and partially compatible settings. Interestingly, in the MHC fully incompatible setting in which only one-third of the recipients demonstrated active thymopoiesis, probably because of the rejection of donor cells by host natural killer cells, IL-7 treatment had a beneficial effect on T-cell development. PMID- 15352990 TI - Prospective clinical evaluation of lower cut-offs for galactomannan detection in adult neutropenic cancer patients and haematological stem cell transplant recipients. AB - The recent advent of an improved commercial serum enzyme-linked immunosorbent assay (ELISA) for the detection of circulating galactomannan (GM), a major constituent of Aspergillus cell walls, has contributed to the diagnosis of invasive aspergillosis (IA) in many haematology and transplant centres. However, the optimal threshold for positivity remains a matter of debate. We prospectively evaluated the impact of lowering the cut-off in 124 neutropenic episodes with a high pretest probability for IA. Two new cut-off points, lower than previously accepted, are proposed: (a) a 'static' cut-off at 0.8 and (b) a 'dynamic' cut-off at 0.5. A single assay with an optical density (OD) index > or = 0.8 warrants the initiation of anti-Aspergillus therapy. A further lowering of the 'static' threshold seems not clinically feasible given the drop in positive predictive value (PPV). However, the demonstration of at least two sequential sera with an OD > or = 0.5 ('dynamic' threshold) increased the specificity and the PPV to 98.6% and the efficiency to 98%. Applying both cut-offs to a subgroup of 21 'possible' fungal infections further identified and upgraded six cases of IA. However, the clinical benefit of lower cut-offs (particularly for earlier diagnosis) depends upon the kinetics of antigenaemia and the intensity of serum sampling. PMID- 15352991 TI - Circulating reticulated platelets in the early and late phases after ischaemic stroke and transient ischaemic attack. AB - The percentage of reticulated platelets (% RP) could be a useful marker of increased platelet production and/or turnover in patients with increased platelet activation, but few flow cytometric studies have measured the % RP in patients with ischaemic cerebrovascular disease (CVD). Whole blood flow cytometry using thiazole orange was performed to compare the % RP in patients in the early (1-27 d, n = 79) and late phases (79-725 d, n = 70) after ischaemic stroke or transient ischaemic attack (TIA) with controls without CVD (n = 27). The impact of aspirin dose escalation (75-300 mg/d) on the % RP was investigated in 10 patients in the late phase after stroke/TIA. The platelet count and mean platelet volume (MPV) were similar in CVD patients and controls. Compared with controls, the unadjusted % RP was not significantly higher in early or late phase CVD patients (P < or = 0.3). However, having adjusted for age, the % RP was higher in early (P = 0.047) and late phase CVD patients (P = 0.01). There was a positive correlation between % RP and MPV in EDTA- and citrate-anticoagulated blood in both early and late phase CVD patients (P< or = 0.01). The % RP was not significantly influenced by aspirin dose. These data do not convincingly support an excessive stimulus to platelet production in the early or late phases after ischaemic stroke/TIA, but are consistent with the hypothesis that reticulated platelets are larger than more mature 'non-reticulated' platelets in ischaemic CVD. PMID- 15352992 TI - Preserved granulocyte formation and function, as well as bone marrow innervation, in subjects with complete spinal cord injury. AB - Patients with a spinal cord injury are at risk of infections and is partly attributed to immobilization. Their lymphocyte-mediated immunity is impaired and the growth of blood progenitor cells is reduced. An adequate immune response depends on granulocytes being mobilized rapidly and activated properly, at the inflammatory site. Possibly this requires a coordinated interaction between the autonomous nervous system and cells within the haematopoietic bone marrow. Granulocyte function in the spinal cord injured has not been evaluated. Although there is evidence that the bone marrow in rodents is innervated, it is uncertain whether human bone marrow is similarly affected. Microscopy and immunolabelling followed by flow cytometry, showed that blood and bone marrow counts of leucocyte subsets were similar in paraplegic, tetraplegic and control subjects (P > 0.05). Neutrophilic migration and oxygen consumption, as well as eosinophil activation, assayed as release of eosinophilic cationic protein or CD69 expression, were not altered after spinal cord injury (P > 0.05). Cryostat sections of human bone marrow biopsies stained positive with glyoxylic acid, indicating the presence of catecholamine-containing nerves in both the patients and the controls. We conclude that terminal differentiation and formation of granulocytes, as well as their functional capacity, do not depend appreciably on supraspinal nervous regulation. PMID- 15352993 TI - Day case management of sickle pain: 3 years experience in a UK sickle cell unit. AB - A day centre was established to determine whether an alternative approach to the management of uncomplicated sickle pain would improve the quality of care and reduce hospital admissions in patients with sickle cell disease. Since the centre opened there has been a 43% decrease in hospital admissions and 49% decrease in occupied bed days. In the third year, 84% of patients treated for severe sickle pain were managed without the need for hospital admission. A centre offering day case management of painful crisis reduced unnecessary hospital admissions for uncomplicated pain. This approach is safe and cost-effective. PMID- 15352994 TI - A novel silent beta-thalassemia mutation in the distal CACCC box affects the binding and responsiveness to EKLF. AB - The silent beta-thalassemia mutation, beta(+)-101C-->T, is the only mutation currently described in the distal beta-globin CACCC box. We present a novel mutation, a C-->G transversion, in the same position. Expression analysis in heterozygous subjects demonstrated that the mutation determines a 20% reduction in the output of the beta-globin gene. DNA-protein interaction and transactivation analysis correlated the decrease in the beta-globin synthesis with the reduced binding and transactivation of EKLF to the mutant promoter. These data predict that the beta-101C-->G mutation will display a silent thalassemia phenotype similar to that of the beta-101C-->T mutation. PMID- 15352995 TI - Effects of anagrelide on platelet factor 4 and vascular endothelial growth factor levels in patients with essential thrombocythemia. PMID- 15352998 TI - The C677T methylenetetrahydrofolate reductase variant and third trimester obstetrical complications in women with unexplained elevations of maternal serum alpha-fetoprotein. AB - INTRODUCTION: The C677T MTHFR variant has been associated with the same third trimester pregnancy complications as seen in women who have elevations of maternal serum alpha-fetoprotein (MSAFP). We hypothesized that these women with third trimester pregnancy complications and MSAFP elevations would have an increased frequency of the variant compared to an abnormal study control group (women with MSAFP elevations without pregnancy complications) as well as to normal population controls. METHODS: Women who had unexplained elevations of MSAFP in pregnancy were ascertained retrospectively. The frequency of the C677T MTHFR variant among those women with unexplained elevations of MSAFP who had experienced later pregnancy complications was compared to that of women with unexplained elevations of MSAFP without complications as well as to that of the previously established Manitoba frequency. RESULTS: Women who had complications of pregnancy and an unexplained MSAFP elevation had a higher allele frequency for the C677T MTHFR variant (q = 0.36,) compared to women with MSAFP elevations and normal pregnancy outcomes (q = 0.25, OR 1.73 95% CI 1.25-2.37, p = 0.03). The frequency was also higher than that of the population controls (q= 0.25, OR 1.70 95% CI 1.11-2.60, p = 0.007). The frequency in women with MSAFP elevations without pregnancy complications was not significantly different from that of the population controls (p = 0.41). CONCLUSION: Women with unexplained elevations of MSAFP and who experience complications in later pregnancy are more likely to have one or two alleles of the C677T MTHFR variant. PMID- 15352999 TI - Glucose-stimulated insulin response in pregnant sheep following acute suppression of plasma non-esterified fatty acid concentrations. AB - BACKGROUND: Elevated non-esterified fatty acids (NEFA) concentrations in non pregnant animals have been reported to decrease pancreatic responsiveness. As ovine gestation advances, maternal insulin concentrations fall and NEFA concentrations increase. Experiments were designed to examine if the pregnancy associated rise in NEFA concentration is associated with a reduced pancreatic sensitivity to glucose in vivo. We investigated the possible relationship of NEFA concentrations in regulating maternal insulin concentrations during ovine pregnancy at three physiological states, non-pregnant, non-lactating (NPNL), 105 and 135 days gestational age (dGA, term 147+/- 3 days). METHODS: The plasma concentrations of insulin, growth hormone (GH) and ovine placental lactogen (oPL) were determined by double antibody radioimmunoassay. Insulin responsiveness to glucose was measured using bolus injection and hyperglycaemic clamp techniques in 15 non-pregnant, non-lactating ewes and in nine pregnant ewes at 105 dGA and near term at 135 dGA. Plasma samples were also collected for hormone determination. In addition to bolus injection glucose and insulin Area Under Curve calculations, the Mean Plasma Glucose Increment, Glucose Infusion Rate and Mean Plasma Insulin Increment and Area Under Curve were determined for the hyperglycaemic clamp procedures. Statistical analysis of data was conducted with Students t-tests, repeated measures ANOVA and 2-way ANOVA. RESULTS: Maternal growth hormone, placental lactogen and NEFA concentrations increased, while basal glucose and insulin concentrations declined with advancing gestation. At 135 dGA following bolus glucose injections, peak insulin concentrations and insulin area under curve (AUC) profiles were significantly reduced in pregnant ewes compared with NPNL control ewes (p < 0.001 and P < 0.001, respectively). In hyperglycaemic clamp studies, while maintaining glucose levels not different from NPNL ewes, pregnant ewes displayed significantly reduced insulin responses and a maintained depressed insulin secretion. In NPNL ewes, 105 and 135 dGA ewes, the Glucose Infusion Rate (GIR) was constant at approximately 5.8 mg glucose/kg/min during the last 40 minutes of the hyperglycaemic clamp and the Mean Plasma Insulin Increment (MPII) was only significantly (p < 0.001) greater in NPNL ewes. Following the clamp, NEFA concentrations were reduced by approximately 60% of pre clamp levels in all groups, though a blunted and suppressed insulin response was maintained in 105 and 135 dGA ewes. CONCLUSIONS: Results suggest that despite an acute suppression of circulating NEFA concentrations during pregnancy, the associated steroids and hormones of pregnancy and possibly NEFA metabolism, may act to maintain a reduced insulin output, thereby sparing glucose for non-insulin dependent placental uptake and ultimately, fetal requirements. PMID- 15353000 TI - Predicting declines in physical function in persons with multiple chronic medical conditions: what we can learn from the medical problem list. AB - BACKGROUND: Primary care physicians are caring for increasing numbers of persons with comorbid chronic illness. Longitudinal information on health outcomes associated with specific chronic conditions may be particularly relevant in caring for these populations. Our objective was to assess the effect of certain comorbid conditions on physical well being over time in a population of persons with chronic medical conditions; and to compare these effects to that of hypertension alone. METHODS: We conducted a secondary analysis of 4-year longitudinal data from the Medical Outcomes Study. A heterogeneous population of 1574 patients with either hypertension alone (referent) or one or more of the following conditions: diabetes, coronary artery disease, congestive heart failure, respiratory illness, musculoskeletal conditions and/or depression were recruited from primary and specialty (endocrinology, cardiology or mental health) practices within HMO and fee-for-service settings in three U.S. cities. We measured categorical change (worse vs. same/better) in the SF-36(R) Health Survey physical component summary score (PCS) over 4 years. We used logistic regression analysis to determine significant differences in longitudinal change in PCS between patients with hypertension alone and those with other comorbid conditions and linear regression analysis to assess the contribution of the explanatory variables. RESULTS: Specific diagnoses of CHF, diabetes and/or chronic respiratory disease; or 4 or more chronic conditions, were predictive of a clinically significant decline in PCS. CONCLUSIONS: Clinical recognition of these specific chronic conditions or 4 or more of a list of chronic conditions may provide an opportunity for proactive clinical decision making to maximize physical functioning in these populations. PMID- 15353001 TI - RNA-binding proteins to assess gene expression states of co-cultivated cells in response to tumor cells. AB - BACKGROUND: Tumors and complex tissues consist of mixtures of communicating cells that differ significantly in their gene expression status. In order to understand how different cell types influence one another's gene expression, it will be necessary to monitor the mRNA profiles of each cell type independently and to dissect the mechanisms that regulate their gene expression outcomes. RESULTS: In order to approach these questions, we have used RNA-binding proteins such as ELAV/Hu, poly (A) binding protein (PABP) and cap-binding protein (eIF-4E) as reporters of gene expression. Here we demonstrate that the epitope-tagged RNA binding protein, PABP, expressed separately in tumor cells and endothelial cells can be used to discriminate their respective mRNA targets from mixtures of these cells without significant mRNA reassortment or exchange. Moreover, using this approach we identify a set of endothelial genes that respond to the presence of co-cultured breast tumor cells. CONCLUSION: RNA-binding proteins can be used as reporters to elucidate components of operational mRNA networks and operons involved in regulating cell-type specific gene expression in tissues and tumors. PMID- 15353002 TI - The effects of moderate alcohol supplementation on estrone sulfate and DHEAS in postmenopausal women in a controlled feeding study. AB - BACKGROUND: We have demonstrated that moderate alcohol consumption (15 g/d, 30 g/d) for 8 weeks resulted in significantly increased levels of serum estrone sulfate and DHEAS in 51 postmenopausal women in a randomized, placebo-controlled trial. We now report on the relationships between serum estrone sulfate and dehydroepiandrosterone sulfate (DHEAS) levels after 4 weeks of moderate alcohol supplementation, and compare the results to the 8 weeks data to elucidate time-to effect differences. METHODS: Postmenopausal women (n = 51) consumed 0 (placebo), 15 (1 drink), and 30 (2 drinks) g alcohol (ethanol)/ day for 8 weeks as part of a controlled diet in a randomized crossover design. Blood samples were drawn at baseline, at 4 weeks and at 8 weeks. Changes in estrone sulfate and DHEAS levels from placebo to 15 g and 30 g alcohol per day were estimated using linear mixed models. RESULTS AND DISCUSSION: At week 4, compared to the placebo, estrone sulfate increased an average 6.9% (P = 0.24) when the women consumed 15 g of alcohol per day, and 22.2% (P = 0.0006) when they consumed 30 g alcohol per day. DHEAS concentrations also increased significantly by an average of 8.0% (P < 0.0001) on 15 g of alcohol per day and 9.2% (P < 0.0001) when 30 g alcohol was consumed per day. Trend tests across doses for both estrone sulfate (P = 0.0006) and DHEAS (P < 0.0001) were significant. We found no significant differences between the absolute levels of serum estrone sulfate at week 4 versus week 8 (P = 0.32) across all doses. However, absolute DHEAS levels increased from week 4 to week 8 (P < 0.0001) at all three dose levels. CONCLUSIONS: These data indicate that the hormonal effects due to moderate alcohol consumption are seen early, within 4 weeks of initiation of ingestion. PMID- 15353003 TI - How patient-physician encounters in critical medical situations affect trust: results of a national survey. AB - BACKGROUND: Patients' trust in physicians and in the medical profession is vital for a successful patient-physician relationship. Trust is especially salient in critical medical situations, such as serious side-effects, hospitalizations, and diagnoses of serious medical conditions, but most trust studies have been done with the general population or in routine primary care settings. This study examines the association between patient-physician encounters in such critical medical situations and patients' trust in their physician and in the medical profession in general. METHODS: A random national telephone survey was conducted using validated multi-item questionnaire measuring trust and satisfaction with physicians and with the medical profession. A seven item questionnaire measured the patient-physician encounters in critical medical situations. A total of 1117 subjects aged 20 years and older with health insurance were included for analyses. Spearman rank order correlations were used to determine the association of encounter variables with trust in physicians and the medical profession. RESULTS: Prescription of medications by primary care physicians that patients believed might have side effects was negatively correlated with trust in physician (rho = -0.12, p < 0.001, n = 1045) in multivariate analysis. A primary care physician evaluating the patient for a condition the patient believed was serious was positively correlated with trust in physician (rho= 0.08, p < 0.01). Being hospitalized was positively correlated with trust in the medical profession (rho = 0.12, p < 0.01, n = 475). CONCLUSION: Hospitalization, perceived seriousness of condition, and concerns about the risks of medications were found to be associated with patient trust in physicians or the medical profession. These findings highlight the salience of trust in serious physician-patient encounters and the role that patient vulnerability plays in determining patient trust. PMID- 15353004 TI - Phenotypic plasticity of fine root growth increases plant productivity in pine seedlings. AB - BACKGROUND: The plastic response of fine roots to a changing environment is suggested to affect the growth and form of a plant. Here we show that the plasticity of fine root growth may increase plant productivity based on an experiment using young seedlings (14-week old) of loblolly pine. We use two contrasting pine ecotypes, "mesic" and "xeric", to investigate the adaptive significance of such a plastic response. RESULTS: The partitioning of biomass to fine roots is observed to reduce with increased nutrient availability. For the "mesic" ecotype, increased stem biomass as a consequence of more nutrients may be primarily due to reduced fine-root biomass partitioning. For the "xeric" ecotype, the favorable influence of the plasticity of fine root growth on stem growth results from increased allocation of biomass to foliage and decreased allocation to fine roots. An evolutionary genetic analysis indicates that the plasticity of fine root growth is inducible, whereas the plasticity of foliage is constitutive. CONCLUSIONS: Results promise to enhance a fundamental understanding of evolutionary changes of tree architecture under domestication and to design sound silvicultural and breeding measures for improving plant productivity. PMID- 15353005 TI - Characterization of a novel large deletion and single point mutations in the BRCA1 gene in a Greek cohort of families with suspected hereditary breast cancer. AB - BACKGROUND: Germline mutations in BRCA1 and BRCA2 predispose to breast and ovarian cancer. A multitude of mutations have been described and are found to be scattered throughout these two large genes. We describe analysis of BRCA1 in 25 individuals from 18 families from a Greek cohort. METHODS: The approach used is based on dHPLC mutation screening of the BRCA1 gene, followed by sequencing of fragments suspected to carry a mutation including intron--exon boundaries. In patients with a strong family history but for whom no mutations were detected, analysis was extended to exons 10 and 11 of the BRCA2 gene, followed by MLPA analysis for screening for large genomic rearrangements. RESULTS: A pathogenic mutation in BRCA1 was identified in 5/18 (27.7 %) families, where four distinct mutations have been observed. Single base putative pathogenic mutations were identified by dHPLC and confirmed by sequence analysis in 4 families: 5382insC (in two families), G1738R, and 5586G > A (in one family each). In addition, 18 unclassified variants and silent polymorphisms were detected including a novel silent polymorphism in exon 11 of the BRCA1 gene. Finally, MLPA revealed deletion of exon 20 of the BRCA1 gene in one family, a deletion that encompasses 3.2 kb of the gene starting 21 bases into exon 20 and extending 3.2 kb into intron 20 and leads to skipping of the entire exon 20. The 3' breakpoint lies within an AluSp repeat but there are no recognizable repeat motifs at the 5' breakpoint implicating a mechanism different to Alu-mediated recombination, responsible for the majority of rearrangements in the BRCA1 gene. CONCLUSIONS: We conclude that a combination of techniques capable of detecting both single base mutations and small insertions/deletions and large genomic rearrangements is necessary in order to accurately analyze the BRCA1 gene in patients at high risk of carrying a germline mutation as determined by their family history. Furthermore, our results suggest that in those families with strong evidence of linkage to the BRCA1 locus in whom no point mutation has been identified re-examination should be carried out searching specifically for genomic rearrangements. PMID- 15353006 TI - Is remission of depressive symptoms in primary care a realistic goal? A meta analysis. AB - BACKGROUND: A primary goal of acute treatment for depression is clinical remission of symptoms. Most meta-analyses of remission rates involve randomized controlled trials (RCTs) using patients from psychiatric settings, but most depressed patients are treated in primary care. The goal of this study was to determine remission rates obtained in RCTs of treatment interventions for Major Depressive Disorder (MDD) conducted in primary care settings. METHODS: Potentially relevant studies were identified using computerized and manual search strategies up to May 2003. Criteria for inclusion included published RCTs with a clear definition of remission using established outcome measures. RESULTS: A total of 13 studies (N = 3202 patients) meeting inclusion criteria were identified. Overall remission rates for active interventions ranged between 50% and 67%, compared to 32% for pill placebo conditions and 35% for usual care conditions. CONCLUSIONS: Remission rates in primary care studies of depression are at least as high as for those in psychiatric settings. It is a realistic goal for family physicians to target remission of symptoms as an optimal outcome for treatment of depression. PMID- 15353007 TI - The FK506 binding protein 13 kDa (FKBP13) interacts with the C-chain of complement C1q. AB - BACKGROUND: The pharmacological action of specific immunosuppressants is mediated by immunophilins. While cyclosporin A binds to cyclophilins, FK506/tacrolimus, rapamycin, and others bind to FK506 binding proteins (FKBPs). Different physiological actions of immunophilins were described but their genuine function, however, remains elusive and is still under investigation. A yeast two-hybrid screen was performed using the FK506 binding protein 13 kDa (FKBP13) as a bait and a fetal liver expression library as a prey. RESULTS: The C-chain of complement C1q (C1q-C) was detected to interact with FKBP13 in the yeast two hybrid system and in a protein complementation assay. Neither FKBP12, FKBP25, FKBP52 nor the unrelated immunophilin CypA did react with C1q-C in the yeast system stressing the specificity of the interaction. Binding of C1q-C to FKBP13 could not be prevented in the presence of FK506, demonstrating that possibly other regions than the binding pocket of the drug are responsible for the interaction of the two proteins. CONCLUSION: It is concluded that exclusively FKBP13 but no other FKBPs tested so far interact with the C-chain of complement C1q in the two different assays and further work will be initiated to investigate the physiological relevance of the interaction. PMID- 15353008 TI - Opioid modulation of GABA release in the rat inferior colliculus. AB - BACKGROUND: The inferior colliculus, which receives almost all ascending and descending auditory signals, plays a crucial role in the processing of auditory information. While the majority of the recorded activities in the inferior colliculus are attributed to GABAergic and glutamatergic signalling, other neurotransmitter systems are expressed in this brain area including opiate peptides and their receptors which may play a modulatory role in neuronal communication. RESULTS: Using a perfusion protocol we demonstrate that morphine can inhibit KCl-induced release of [3H]GABA from rat inferior colliculus slices. DAMGO ([D-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin) but not DADLE ([D-Ala2, D Leu5]-enkephalin or U69593 has the same effect as morphine indicating that micro rather than delta or kappa opioid receptors mediate this action. [3H]GABA release was diminished by 16%, and this was not altered by the protein kinase C inhibitor bisindolylmaleimide I. Immunostaining of inferior colliculus cryosections shows extensive staining for glutamic acid decarboxylase, more limited staining for micro opiate receptors and relatively few neurons co-stained for both proteins. CONCLUSION: The results suggest that micro-opioid receptor ligands can modify neurotransmitter release in a sub population of GABAergic neurons of the inferior colliculus. This could have important physiological implications in the processing of hearing information and/or other functions attributed to the inferior colliculus such as audiogenic seizures and aversive behaviour. PMID- 15353009 TI - Evaluating integrative medicine: science and art. Commentary on "A descriptive analysis of an integrative medicine clinic". PMID- 15353010 TI - From a healer to scientists: on duality. PMID- 15353011 TI - Comments on the energy issue. PMID- 15353012 TI - Probiotics in the prevention and treatment of diarrhea. PMID- 15353013 TI - Cranberries and urinary-tract health: a knowledge assessment of American College of Obstetricians and Gynecologists fellows. PMID- 15353014 TI - Can relaxation programs with music enhance human immune function? PMID- 15353015 TI - Healing vision without spectacles, contact lenses or refractive surgery: a personal journey of healing vision. PMID- 15353016 TI - Reliability of the Prognos electrodermal device for measurements of electrical skin resistance at acupuncture points. AB - OBJECTIVES: (1) To characterize and calibrate an electrodermal screening device, Prognos. (2) To replicate a previous test-retest reliability study of this device with measurements of electrical skin resistance (ESR) at 24 Jing-well acupuncture points (APs). (3) To determine measurement precision in three successively more exacting trial protocols on the same set of subjects. SETTINGS: Oregon College of Oriental Medicine and Portland State University, Portland, OR. INSTRUMENTS: The Prognos device was electrically characterized by a team of research engineers at the Biomedical Signal Processing Laboratory of Portland State University. They determined that Prognos measures the average direct-current (DC) resistance between a metallic wrist strap and an electrode probe tip. The probe tip is connected to a linear spring set to trigger with an optically generated signal at a deflection of 2.62 mm, which corresponds to an average applied force of 2.68 +/ 0.04 N (mean +/- standard deviation [SD], n = 6). They also determined that the device quantifies resistance by applying a 1.1 microA current for an average of 223 +/- 3 ms (n = 7). When calibrated against a series of known resistors, Prognos measures accurately in the range of 150 kOmega to 14.3 MOmega with an error of less than 0.4%. SUBJECTS: Thirty-one (31) healthy volunteers, 17 females and 14 males, 23-63 years of age. RESULTS OF RELIABILITY TEST-RETEST: The mean reliability of a single measurement was; 0.758 for a standard measurement protocol of four sequential sweeps of 24 Jing-well (Ting) APs; 0.851 for four sequential sweeps after ink-marking the APs; and 0.961 for four rapid repeat measurements at each inked AP. Mean absolute values of ESR decreased between the standard and marked protocols, but not between the marked and rapid repeat protocols. CONCLUSIONS: Prognos performs accurately, against known resistors over the reported range of ESR. The reliability in the standard protocol (r = 0.758) is comparable to the reliability of 0.721 demonstrated under similar conditions by other investigators. Marking APs, and performing measurements in a rapid sequence, increases reliability of ESR measurements. Increased reliability in the second and third protocols is associated with decreased mean ESR values which may be related to increased accuracy of Prognos probe placement and/or inking the APs. PMID- 15353017 TI - Surface electromyography apparatus as a measurement device for biofield research: results from a single case study. AB - OBJECTIVES: The objective of this preliminary investigation was to determine if surface electromyography (EMG) equipment is capable of detecting electrical signals that can be used as a measure in biofield research. DESIGN: These preliminary data were collected from one subject (a member of the research team), using the subject as his own control. Resting state data are compared to data collected during a specific Healing Touch (HT) intervention. SETTINGS/LOCATION: The setting was in an electromagnetically shielded room (a modular enclosure utilizing two separate conductive electromagnetic barriers, which were electrically isolated, attached by a single-point ground connection) at the University of Wisconsin-Milwaukee, Milwaukee, WI. INTERVENTION: Data were collected during a resting state, and subsequently during a partial "HT back sequence." The back sequence is a standard HT procedure consisting of "Connecting the Lower Body," "Open Spinal Energy Flow," a "Vertebral Spiral Technique," and other techniques taught in HT level 2. MEASURES: Electrodes were attached to the back of the subject at eight acupoints. The Model 544 EMG System (amplifier and electrode assembly) (Therapeutics Unlimited, Inc. Iowa City, IA) was used to acquire data. The raw data were divided into discrete time segments and converted from the time domain into the frequency domain by performing a fast Fourier transform. Integer frequency averaging was conducted to allow for ease of interpretation. Using the voltages at each frequency, the signal strength was calculated in decibels. RESULTS: A 0-65 Hz frequency spectrum was obtained from the surface EMG apparatus. An averaged baseline, "resting state" frequency spectrum was definable for the subject that was stable with time for signal strengths >0 dB. When the HT practitioner attuned with the subject, higher frequency components of the spectrum increased in amplitude, peaked, then decreased throughout the intervention. CONCLUSIONS: The conclusions drawn from these results is that the surface EMG apparatus has potential as a measurement device for biofield research. Further experimentation using this device for biofield measurement is encouraged. PMID- 15353018 TI - Effectiveness gaps: a new concept for evaluating health service and research needs applied to complementary and alternative medicine. AB - BACKGROUND: An effectiveness gap (EG) is an area of clinical practice in which available treatments are not fully effective. EGs have not been previously researched. Complementary and alternative medicine (CAM) interventions, by definition, are not generally available through normal health care channels. Therefore, if effective, they have the potential to increase achieved community effectiveness. AIMS: A pilot study to determine whether EGs exist, and if so to provide initial data on their nature, frequency, and causes. To obtain preliminary data on whether CAM may offer effective interventions in these clinical areas. DESIGN: Semistructured telephone interviews; literature search. SETTING: Twenty-two (22) general practitioners (GPs) in London, U.K. METHOD: One hundred and fifty-two (152) doctors who had responded to an earlier survey on attitudes to CAM were approached. Respondents were asked to specify EGs and to give reasons why available treatment is unsatisfactory and to estimate the frequency and severity of clinical problems relating to EGs. Sampling was continued to redundancy. A bibliometric study examined the volume and type of published evidence on the effectiveness of CAM interventions in the identified clinical areas. RESULTS: There was good concordance among respondents on EGs encountered in general/family practice. Seventy-eight (78) clinical problems were cited. EGs are encountered quite frequently: 68 of 78 (85%) of EGs were encountered at least once per month. Musculoskeletal problems were cited by 20 of 22 (90%) of respondents as being affected by EGs. Depression, eczema, chronic pain, and irritable bowel syndrome were also frequently mentioned. Systematic reviews and meta-analyses conclude that there is evidence for the effectiveness of various CAM interventions in most of these areas. CONCLUSIONS: EGs, mapped against evidence, have the potential to inform service development and research policy. Further study should be undertaken: it should incorporate improved sampling and data collection methodology. Specifically, where effective CAM interventions exist but are not being applied, EGs form part of the "avoidable burden of illness" identified by early work on evidence-based medicine. Practice guidelines should incorporate CAM interventions where there is evidence. The CAM research agenda should focus on areas affected by EGs. PMID- 15353019 TI - Dietary salt intake as a potential modifier of airway responsiveness in bronchial asthma. AB - While pharmacologic treatment of chronic asthma is usually highly effective, medications often have significant side-effects or exhibit tachyphylaxis. Alternative and/or complementary treatments that reduce dependence on pharmacologic medications are of interest in reducing the severity of asthma. This review analyzes the literature that has evaluated dietary salt intake as a potential modifier of the severity of asthma and airway responsiveness. High dietary intakes of salt, greater than 9 g/d, are common in Western civilizations, as is asthma. The question is whether reducing dietary salt intake potentially would improve pulmonary function and airway responsiveness in individuals with asthma. This review details the existing studies in this regard and includes the studies that have evaluated dietary salt on the severity of exercise-induced asthma (exercise-induced bronchoconstriction [E1B]). From a critical analysis of the existing literature, the data that support a role for dietary salt reduction for reducing severity of asthma and airway responsiveness in individuals with asthma is considered encouraging but not clinically convincing. The existing studies have suffered from a variety of experimental and population limitations. In contrast, the data from studies that have altered dietary salt and evaluated severity of EIB in nonatopic individuals is much more convincing. In each study so far, lowering dietary salt has reduced the severity of EIB to subclinical levels. Correspondingly, the supplementing of diets to higher than normal salt intake increased EIB significantly. This review concludes that the data are sufficient to warrant a clinical trial that is properly controlled and randomized to further investigate the influence of dietary salt intake on pulmonary function, airway responsiveness, symptoms, quality of life, and medication requirements in asthma and EIB. PMID- 15353020 TI - New diagnostic and therapeutic approach to thyroid-associated orbitopathy based on applied kinesiology and homeopathic therapy. AB - OBJECTIVES: To investigate pathogenetic mechanisms related to the lacrimal and lymphatic glands in patients with thyroid-associated orbitopathy (TAO), and the potential of applied kinesiology diagnosis and homeopathic therapeutic measures. DESIGN: Prospective. SETTINGS/LOCATION: Thyroid outpatient unit and a specialized center for complementary medicine (WOMED, Innsbruck; R.M. and H.M.). SUBJECTS: Thirty-two (32) patients with TAO, 23 with a long-standing disease, and 9 showing discrete initial changes. All patients were euthyroid at the time of the investigation. INTERVENTIONS: Clinical investigation was done, using applied kinesiology methods. Departing from normal reacting muscles, both target organs as well as therapeutic measures were tested. Affected organs will produce a therapy localization (TL) that turns a normal muscle tone weak. Using the same approach, specific counteracting therapies (i.e., tonsillitis nosode and lymph mobilizing agents) were tested. OUTCOME MEASURES: Change of lid swelling, of ocular movement discomfort, ocular lock, tonsil reactivity and Traditional Chinese Medicine criteria including tenderness of San Yin Jiao (SP6) and tongue diagnosis were recorded in a graded fashion. RESULTS: Positive TL reactions were found in the submandibular tonsillar structures, the pharyngeal tonsils, the San Yin Jiao point, the lacrimal gland, and with the functional ocular lock test. Both Lymphdiaral (Pascoe, Giessen, Germany) and the homeopathic preparation chronic tonsillitis nosode at a C3 potency (Spagyra, Grodig, Austria) counteracted these changes. Both agents were used therapeutically over 3-6 months, after which all relevant parameters showed improvement. CONCLUSIONS: Our study demonstrates the involvement of lymphatic structures and flow in the pathogenesis of TAO. The tenderness of the San Yin Jiao point correlates to the above mentioned changes and should be included in the clinical evaluation of these patients. PMID- 15353021 TI - A descriptive analysis of an integrative medicine clinic. AB - OBJECTIVE: To evaluate and describe an integrative medicine clinic including its patients; their presenting problems, medical objectives, treatment recommendations; whether recommendations were recalled and adhered to; and patients' self-reported health outcomes. DESIGN: A prospective cohort study of 160 new clinic patients were treated using a wide range of integrative medicine therapies. Patients were interviewed at intervals of 1, 3, and 6 months after their initial visit. SETTING: The study was conducted at the Institute for Health and Healing clinic, at California Pacific Medical Center located in San Francisco, CA. SUBJECTS: The subjects were new clinic patients seeking care for a variety of symptoms and diagnoses. INTERVENTION: Patients presented with an average of 4.0 symptoms and 2.9 diagnoses; physicians recommended 9.5 "treatments" per patient. RESULTS: At 6-month follow-up, symptom intensity decreased from 6.5 to 4.2 (on a 10-point scale; p < 0.005); and 57% of the patients had made "substantial" progress or had completely achieved their health objectives (p < 0.005). The Short Form 12 (SF-12) measure of mental and physical functioning improved significantly from baseline to one month (p = 0.05; p = 0.001, respectively) and maintained this improvement at 3 (p = 0.01; p = 0.01) and 6 months (p = 0.001; p = .001). At 1-month follow-up, patients recalled 57% of the actionable statements from the physician without a prompt and they followed 55% of all recommendations well (90% or above). Over the follow-up period, patients increasingly attributed the reductions in symptoms (p = 0.01) and achieving their health objectives (p = 0.01) to the treatment plan. Self reported measures of days ill and days missed at work/school did not differ significantly from the year before coming to the clinic. CONCLUSION: The results indicate that patients can recall and follow a complex treatment regimen. Subjective and objective measures of patients' health status improved after one month and this effect was sustained or improved further at 6 months follow-up. Further study is needed to determine whether patients' improvement was the result of the treatment regimen. PMID- 15353022 TI - Supplement use among cancer survivors in the Vitamins and Lifestyle (VITAL) study cohort. AB - OBJECTIVES: Although dietary supplement use is believed to be prevalent among patients with cancer, less is known about specific associations of supplements with types of cancer. These analyses describe the use of specific supplements by 10,857 cancer survivors in comparison to cancer-free controls (n = 64,226). DESIGN: Cross-sectional cohort study. SUBJECTS/SETTINGS/LOCATION: Participants were members of the VITAL (Vitamins and Lifestyle) study, a cohort study of 75,288 men and women in western Washington State, 50-76 years of age, examining supplement use and future cancer risk. INTERVENTIONS/OUTCOME MEASURES: Respondents completed a 24-page questionnaire, including information on cancer history, demographic characteristics, and current use of vitamin, mineral, and herbal supplements. RESULTS: In this cohort, cancer survivors used similar numbers of supplements as cancer-free controls. Analyses examined the association of a diagnosis history of 11 specific cancers (n > or = 280), by anatomic site, with the high use of multivitamins and 16 individual vitamin/mineral and 20 herbal/specialty supplements. In analyses adjusted for gender, age, education, and race/ethnicity, odds ratios and 95% confidence intervals for the strongest positive associations were: cranberry pills with bladder cancer 3.44 (1.86, 6.35); zinc with ovarian cancer 2.19 (1.41, 3.40); soy with prostate cancer 1.99 (1.38, 2.87); melatonin with cervical cancer 1.86 (1.19, 2.90); and vitamin D with thyroid cancer 1.66 (1.21, 2.28). CONCLUSIONS: Future research needs to assess the efficacy and safety of these specific supplements in these specific cancer populations. PMID- 15353023 TI - Artichoke leaf extract reduces symptoms of irritable bowel syndrome and improves quality of life in otherwise healthy volunteers suffering from concomitant dyspepsia: a subset analysis. AB - OBJECTIVES: Does artichoke leaf extract (ALE) ameliorate symptoms of Irritable bowel syndrome (IBS) in otherwise healthy volunteers suffering concomitant dyspepsia? METHODS: A subset analysis of a previous dose-ranging, open, postal study, in adults suffering dyspepsia. Two hundred and eight (208) adults were identified post hoc as suffering with IBS. IBS incidence, self-reported usual bowel pattern, and the Nepean Dyspepsia Index (NDI) were compared before and after a 2-month intervention period. RESULTS: There was a significant fall in IBS incidence of 26.4% (p < 0.001) after treatment. A significant shift in self reported usual bowel pattern away from "alternating constipation/diarrhea" toward "normal" (p < 0.001) was observed. NDI total symptom score significantly decreased by 41% (p < 0.001) after treatment. Similarly, there was a significant 20% improvement in the NDI total quality-of-life (QOL) score in the subset after treatment. CONCLUSION: This report supports previous findings that ALE ameliorates symptoms of IBS, plus improves health-related QOL. PMID- 15353024 TI - Retrospective case series on patients with chronic spinal pain treated with dextrose prolotherapy. AB - OBJECTIVES: To determine the clinical benefits of dextrose prolotherapy in patients with chronic spinal pain. DESIGN: Retrospective case series. SETTING/LOCATION: During the first 2 years at an outpatient prolotherapy clinic. SUBJECTS: One hundred and seventy-seven (177) consecutive patients with a history of chronic spinal pain completed prolotherapy treatment and were followed for a period ranging from 2 months to 2.5 years. INTERVENTIONS: Patients were treated with a proliferant solution containing 20% dextrose and 0.75% xylocaine. One half milliliter (0.5 mL) of proliferant was injected into the facet capsules of the cervical, thoracic, and lumbar spine, or combinations of the three areas. The iliolumbar and dorsal sacroiliac ligaments were also injected in patient with low back pain. Injections were typically done on a weekly basis for up to 3 weeks. A set of three injections was repeated in 1 month's time if needed. OUTCOME MEASURES: Level of pain, and improvement in activities of daily living were measured on a five-point scale. Improvement in ability to work was also assessed. RESULTS: Ninety-one percent (91.0%) of patients reported reduction in level of pain; 84.8% of patients reported improvement in activities of daily living, and 84.3% reported an improvement in ability to work. Women required on average, three more injections than men. Cervical spine response rates were lower than thoracic or lumbar spine. No complications from treatment were noted. CONCLUSIONS: Dextrose prolotherapy appears to be a safe and effective method for treating chronic spinal pain that merits further investigation. Future studies need to consider differences in gender response rates. PMID- 15353025 TI - Impact of qigong exercise on self-efficacy and other cognitive perceptual variables in patients with essential hypertension. AB - OBJECTIVES: The purpose of this study was to investigate the impact of practicing qigong on middle-age subjects with essential hypertension. Impacts on blood pressure, reported self-efficacy, perceived benefit, and emotion were observed. DESIGN: Thirty-six (36) adult volunteers were assigned to either a waiting list control or a qigong group that practiced two 30-minute qigong programs per week over 8 consecutive weeks. RESULTS: Systolic and diastolic blood pressure was significantly reduced in members of the qigong group after 8 weeks of exercise. Significant improvements in self-efficacy and other cognitive perceptual efficacy variables were also documented in the qigong group compared to the original situation described above. CONCLUSIONS: This pilot study demonstrates the positive effects of practicing qigong on controlling blood pressure and enhancing perceptions of self-efficacy. PMID- 15353026 TI - Effect of qi training on neutrophil function in young and elderly males. AB - OBJECTIVES: To examine the effect of qi training on neutrophil bactericidal function (superoxide generation and adhesion). METHODS: We studied the effects of one session of qi training on superoxide generation and adhesion of neutrophils immediately after (Post I), and 2 hours after qi training (Post II), in nine young and nine elderly male subjects. RESULTS: The qi training significantly enhanced the superoxide generation and adhesion of neutrophils, and there were significant differences at Post I compared to before qi training (Pre) in both groups. CONCLUSION: Our current observations show that qi training enhances superoxide generation and adhesion of neutrophils. It is supposed that qi training may increase the resistance of trained individuals against common infection and inflammation. PMID- 15353027 TI - Washing hands before and after performing massages? Changes in bacterial survival count on skin of a massage therapist and a client during massage therapy. AB - OBJECTIVE: Massage therapy involves direct contact between the therapist and the client in a clinical setting, which can result in the transfer of bacteria. The purpose of this study was to determine the degree of bacteria transfer between a therapist and a client during a massage session and differences between types of lubricants used. DESIGN: Medium cultures were taken from the therapist's palms and the client's skin before, during, and after the massage session. Experiments proceeded with the following settings: use of three different types of lubricants, without the use of a lubricant, and without massage as control. After each massage session, the therapist washed his or her hands and a bacterial sample was again taken. MAIN OUTCOME MEASURES: Changes in bacteria count. RESULTS: (1) Bacteria count on the therapist's palms increased during and after massage with and without the use of lubricant. There was no change in the case of no massage. (2) Bacteria count on the client's skin decreased during and after massage with the use of three different kinds of lubricants compared to the no massage control. However, an increase was evident when no lubricant was used. (3) After hand washing with water for 20 seconds after each massage session, there were still bacteria on the therapist's palms. However, bacteria count differed with the type of lubricant used. CONCLUSION: Bacteria from the client's skin transferred to the therapist's hands during massage therapy. Bacteria count when no lubricant was used was greater than massage sessions using lubricants. PMID- 15353028 TI - Inhibition of prostate cancer-cell proliferation by Essiac. AB - OBJECTIVE: To assess the ability of Essiac tea extracts (Essiac Canada International, Ottawa, Canada) to modulate cancer cell proliferation and immune responsiveness. DESIGN: A noncancerous transformed cell line was compared to a cancerous cell line and spleen cells that had been isolated from mice to examine proliferation responses mediated by the addition of an Essiac preparation. RESULTS: We found in vitro evidence of decreased proliferation of both noncancerous transformed (CHO) and cancerous prostate cell line (LNCaP) when Essiac was present in the culture media. A dose response for inhibition was demonstrated by a linear regression performed on the data for both the CHO and LNCaP cells. The percent inhibition of the LNCaP cells was higher than the percent inhibition of the CHO cells suggesting that Essiac may have a more selective effect on cancer cells than transformed cells. In addition, the effects of Essiac were examined in an immune T-lymphocyte proliferation assay. At low doses of Essiac, augmentation of proliferation of these T cells was demonstrated, but at higher doses Essiac was inhibitory to T-cell proliferation. The same doses of Essiac that stimulated spleen cells were inhibitory for LNCaP cell proliferation. CONCLUSIONS: Essiac preparations may be able to inhibit tumor cell growth while enhancing immune response to antigenic stimulation. This may be especially valuable in immune-suppressed individuals. PMID- 15353029 TI - A brief history of time: the power of botanical systematic reviews. AB - There are more than 20 completed Cochrane systematic reviews on botanical medicine presently published in the Cochrane Library. There are more than 40 that are planned or in progress. It is an opportune time to explore the information needs of readers of botanical systematic reviews and how those needs can be met better by Cochrane systematic reviews. It is proposed that Cochrane systematic reviews focus not only on efficacy but also on expanded safety and quality. Expanded safety refers not only to the occurrence of adverse events but also the contraindications for use such as drug-herb interactions or allergies to products. Quality pertains to whether or not there was a method of standardizing active ingredients in trials and methods for minimizing risks of contamination. Because there are no package inserts to accompany herbal products as there are for drugs, Cochrane systematic reviews offer the ideal forum to present this much needed information on expanded safety and quality. PMID- 15353030 TI - One size does not fit all: aptitude chi treatment interaction (ATI) as a conceptual framework for complementary and alternative medicine outcome research. Part II--research designs and their applications. AB - When multiple treatment choices are available, the question is not just "which treatment is the best?" but more importantly "best or better for whom, when, and why?" Aptitude (or attribute) by treatment interaction (ATI) is a research paradigm that attempts to examine exactly that--how outcome depends on the match or mismatch between patients' specific characteristics and the treatments they receive. The purpose of this two-part paper is to introduce ATI methods as a conceptual framework into complementary and Alternative medicine/integrative medicine (CAM/IM) outcome research. Part I presented key concepts in ATI research. Part II presents ATI research designs and discusses their applications to the examination of the relationships between individuals and therapies, and the illumination of the mechanisms that make therapies differentially effective. Based on this examination, we conclude that ATI research offers invaluable insights into the multifaceted package of care typically delivered in contemporary medicine and therefore should be included in the portfolio of all CAM/IM outcome research. PMID- 15353031 TI - Integrating Chinese traditional medicine into a U.S. public health paradigm. AB - Chinese traditional medicine (CTM) is a health care system with an extensive history of practical clinical experience. The foundation of CTM, while relatively simple, is substantively different from much of allopathic medicine. Such differences are difficult to explain using a Western medical vocabulary, and extend beyond linguistic foundations. This proves challenging when trying to identify appropriate teaching and research methods that are sensitive to the CTM paradigm and yet relevant to a public health orientation. Given the increased use of CTM, it becomes important to address possibilities that would ensure successful integration of CTM into a public health framework. We propose a model in which both CTM and biomedical clinical services could be offered to provide diverse, yet truly integrative, therapeutic approaches. Within this model, it is critical to enhance reciprocal educational and research-directed opportunities for both CTM practitioners and allopathic clinicians. Considerable responsibility rests upon academic institutions in becoming proactive in developing and implementing educational curricula and research programs that illustrate more effectively the potential bilateral benefit(s), limitations, and, ultimately, roles that CTM and biomedical approaches may assume within an integrative system of care. PMID- 15353037 TI - Vesicular stomatitis virus: a potential therapeutic virus for the treatment of hematologic malignancy. AB - Certain strains of vesicular stomatitis virus (VSV) have been shown to be oncolytic in a wide variety of solid tumors. In the present study, we tested the leukemolytic properties of VSV using established leukemia cell lines and primary patient material. VSV efficiently killed essentially all leukemic cell lines. In contrast, however, normal clonogenic bone marrow progenitor cells and peripheral blood cells were remarkably refractory to infection by VSV. By exploiting this large difference in susceptibility to infection we successfully purged contaminating leukemic cells from cultures of peripheral blood progenitor cells (PBPC) using VSV. VSV was also able to infect and kill leukemic cells in primary samples taken from patients with multiple myeloma (MM). This study demonstrates the potential utility of VSV in the treatment, both ex vivo and in vivo, of hematologic malignancies. PMID- 15353038 TI - Functional characterization of a recombinant adeno-associated virus 5-pseudotyped cystic fibrosis transmembrane conductance regulator vector. AB - Despite extensive experience with recombinant adeno-associated virus (rAAV) 2 vectors in the lung, gene expression has been low in the context of cystic fibrosis (CF) gene therapy, where the large size of the cystic fibrosis transmembrane conductance regulator (CFTR) coding sequence has prompted the use of compact endogenous promoter elements. We evaluated the possibility that gene expression from recombinant adeno-associated virus (rAAV) could be improved by using alternate AAV capsid serotypes that target different cell-surface receptors (i.e., rAAV5) and/or using stronger promoters. The relative activities of the cytomegalovirus (CMV) Rous sarcoma virus (RSV) promoter, the CMV enhancer/beta actin (CB) promoter combination, and the CMV enhancer/RSV promoter hybrid were assessed in vitro in a CF bronchial cell line. The CB promoter was the most efficient. AAV capsid serotypes, rAAV2 and rAAV5, were also compared, and rAAV5 was found to be significantly more efficient. Based on these studies a rAAV5-CB promoter-driven CFTR minigene vector was then used to correct the CF chloride transport defect in vitro, as well as the hyperinflammatory lung phenotype in Pseudomonas-agarose bead challenged CF mouse lungs in vivo. These studies provide functional characterization of a new version of rAAV-CFTR vectors. PMID- 15353039 TI - Modulation of eomes activity alters the size of the developing heart: implications for in utero cardiac gene therapy. AB - Congenital heart disease is the most prevalent cause of infant morbidity and mortality in developed countries. The mechanisms responsible for many specific types of congenital cardiac malformations are strongly associated with gene abnormalities. However, at this time no strategies for gene therapy of the various congenital heart malformations have been investigated. In the present studies we focus on Eomesodermin (Eomes), a T-box transcription factor expressed in developing vertebrate mesoderm. Although Eomes is required for early mesodermal patterning and differentiation, the role of Eomes in cardiac development is unknown. In the present studies we demonstrate that Eomes is expressed in the developing heart, with a pronounced myocardial distribution in the Xenopus ventricle during late cardiac development. Using either a conditional dominant-interfering approach (GR-Eomes--engrailed) or an Eomes-activating approach (GR-Eomes-VP16) we demonstrate that manipulating Eomes activity during late cardiac development can either suppress ventricular development (GR-Eomes enR) or increase ventricular myocardial size (GR-Eomes-VP16). Thus, a potential gene therapy approach for treating both congenital ventricular hypoplasia (e.g., the hypoplastic left heart syndrome) and hypertrophic cardiomyopathy is hypothetically implicit from the present results. PMID- 15353040 TI - Augmentation of antitumor activity of a recombinant adeno-associated virus carcinoembryonic antigen vaccine with plasmid adjuvant. AB - Recombinant adeno-associated virus 2 (rAAV) vectors have been successfully used for sustained expression of therapeutic genes. The potential of using rAAV as a cancer vaccine vector and the impact of a bacterial plasmid adjuvant on this activity were investigated. C57BL/6 mice received a single intramuscular injection of rAAV expressing the human tumor-associated antigen, carcinoembryonic antigen (CEA). Three weeks later, when CEA expression was optimal, a bacterial plasmid containing methylated DNA motifs was injected into the same muscle. Mice were challenged 1 week later with syngeneic MC38 tumor cells stably expressing CEA. Immunization with rAAV-CEA alone resulted in sustained transgene expression and the elicitation of a humoral immune response to CEA. Cellular immune response, however, was weak, and tumor protection was not significant. In contrast, immunization with rAAV-CEA and the plasmid adjuvant resulted in stronger cellular immune response to CEA and tumor protection. The addition of plasmid adjuvant increased both myeloid dendritic cell recruitment in situ and CEA-specific T-helper-1-associated immune response. These data indicate that robust rAAV transgene expression of a tumor antigen followed by transient plasmid delivery to recruit and activate dendritic cells is an effective method of eliciting antitumor cellular immune responses. PMID- 15353041 TI - A synthetic Rev-independent bovine immunodeficiency virus-based packaging construct. AB - Replication competent lentivirus (RCL) has been the major safety concern associated with applications of lentivirus-based gene transfer systems for human gene therapy. Minimization and elimination of overlaps between the packaging and the transfer vector constructs are expected to reduce the potential to generate RCL. We previously developed second- and third-generation bovine immunodeficiency virus (BIV)-based gene transfer systems. However, some sequence homologies between the vector and gag/pol packaging constructs remained. In order to minimize the sequence homologies, we recoded gag/pol with codon usage optimized for expression in human cells in this report. Expression of the recoded gag/pol was Rev/RRE independent. Thus, RRE was eliminated from the packaging construct, thereby removing a 312 bp block of homology. In addition, recoding gag/pol minimized overall homologies between the packaging and transfer vector constructs. Vectors generated by the recoded packaging construct with a four plasmid system had titers greater than 1 x 10(6) transducing units per milliliter, equivalent to those of the earlier generation systems. The vectors were functional in vitro and efficiently transduced rat pigment epithelial cells in vivo. Generation of the synthetic packaging construct provides further advances to the safety of lentiviral vectors for clinical applications. PMID- 15353042 TI - Protective immunity against trypanosoma cruzi infection in a highly susceptible mouse strain after vaccination with genes encoding the amastigote surface protein 2 and trans-sialidase. AB - Protective immunity against lethal infection is developed when BALB/c or C57BL/6 mice are immunized with plasmids containing genes from the protozoan parasite Trypanosoma cruzi. However, genetic vaccination of the highly susceptible mouse strain A/Sn promoted limited survival after challenge. This observation questioned whether this type of vaccination would be appropriate for highly susceptible individuals. Here, we compared the protective efficacy and the immune response after individual or combined genetic vaccination of A/Sn mice with genes encoding trans-sialidase (TS) or the amastigote surface protein-2 (ASP-2). After challenge, a significant proportion of A/Sn mice immunized with either the asp-2 gene or simultaneously with asp-2 and ts genes, survived infection. In contrast, the vast majority of mice immunized with the ts gene or the vector alone died. Parasitological and histological studies performed in the surviving mice revealed that these mice harbored parasites; however, minimal inflammatory responses were seen in heart and striated muscle. We used this model to search for an in vitro correlation for protection. We found that protective immunity correlated with a higher secretion of interferon- by spleen cells on in vitro restimulation with ASP-2 and the presence of ASP-2-specific CD8 cells. Depletion of either CD4 or CD8 or both T-cell subpopulations prior to the challenge rendered the mice susceptible to infection demonstrating the critical contribution of both cell types in protective immunity. Our results reinforce the prophylactic potential of genetic vaccination with asp-2 and ts genes by describing protective immunity against lethal T. cruzi infection and chronic tissue pathology in a highly susceptible mouse strain. PMID- 15353043 TI - Functional expression of the single subunit NADH dehydrogenase in mitochondria in vivo: a potential therapy for complex I deficiencies. AB - It has been reported that defects of mitochondrial proton-translocating NADH quinone oxidoreductase (complex I) are involved in many human diseases (such as encephalomyopathies and sporadic Parkinson's disease). However, no effective remedies have been established for complex I deficiencies. We have adopted a gene therapy approach utilizing the NDI1 gene that codes for the single subunit NADH dehydrogenase of Saccharomyces cerevisiae (Ndi1). Our earlier experiments show that the Ndi1 protein can replace or supplement the functionality of complex I in various cultured cells. For this approach to be useful, it is important to demonstrate in vivo that the mature protein is correctly placed in mitochondria. In this study, we have attempted in vivo expression of the NDI1 gene in skeletal muscles and brains (substantia nigra and striatum) of rodents. In all tissues tested, the Ndi1 protein was identified in the injected area by immunohistochemical staining at 1-2 weeks after the injection. Sustained expression was observed for at least 7 months. Double-staining of the sections using antibodies against Ndi1 and F(1)-ATPase revealed that the expressed Ndi1 protein was predominantly localized to mitochondria. In addition, the tissue cells expressing the Ndi1 protein stimulated the NADH dehydrogenase activity, suggesting that the expressed Ndi1 is functionally active. It was also confirmed that the Ndi1 expression induced no inflammatory response in the tissues examined. The data indicate that the NDI1 gene will be a promising therapeutic tool in the treatment of encephalomyopathies and neurodegenerative diseases caused by complex I impairments. PMID- 15353044 TI - Trans-splicing adeno-associated viral vector-mediated gene therapy is limited by the accumulation of spliced mRNA but not by dual vector coinfection efficiency. AB - Therapeutic application of recombinant adeno-associated virus (AAV) has been limited by its small carrying capacity. To overcome this limitation trans splicing vectors were developed recently. However, the transduction efficiency of trans-splicing vectors is considerably lower than that of a single intact vector in skeletal muscle. To improve trans-splicing vectors for skeletal muscle gene therapy, we examined whether coinfection efficiency is a rate-limiting factor in the mdx mouse, a model for Duchenne muscular dystrophy. Two different AAV viruses were delivered to the mdx muscle. Similar to previous reports in normal muscle, coinfection efficiency reached approximately 90% in the diseased muscle. This result suggests that coinfection is not a hurdle in dystrophic muscle. Another critical step in the trans-splicing method is the transcription and splicing across the inverted terminal repeat (ITR) junction in the reconstituted genome. To test whether this represented a significant obstacle, we systematically evaluated the transcription, pre-mRNA stability and splicing, and translation in a synthetic lacZ construct that mimicked the reconstituted genome. Although inserting an intron in the lacZ gene had no effect on its expression, inclusion of the ITR junction in the intron reduced expression by 50%. In construct containing the ITR junction, the mRNA transcript level was significantly reduced. This mRNA level reduction was associated with decreased pre-mRNA stability. These data suggest that the accumulation of mRNA is a rate-limiting factor in trans splicing vector-mediated gene therapy. PMID- 15353045 TI - Long-term correction of murine lipoprotein lipase deficiency with AAV1-mediated gene transfer of the naturally occurring LPL(S447X) beneficial mutation. AB - Human lipoprotein lipase (LPL) deficiency causes profound hypertriglyceridemia and life-threatening pancreatitis. We recently developed an adult murine model for LPL deficiency: LPL -/- mice display grossly elevated plasma triglyceride (TG) levels (>200-fold) and very low high-density lipoprotein cholesterol (HDL-C < 10% of normal). We used this animal model to test the efficacy of adeno associated virus-mediated expression of hLPL(S447X) (AAV1-LPL(S447X)) in muscle for the treatment of LPL deficiency. Intramuscular administration of AAV1 LPL(S447X) resulted in dose-dependent expression of hLPL protein and LPL activity (up to 33% of normal murine levels) in postheparin plasma. Remarkably, visible hyperlipidemia was resolved within 1 week; plasma TG was reduced to near-normal levels (from 99.0 to 1.8 mmol/L), and plasma HDL-C was increased 6-fold (from 0.2 to 1.1 mmol/L). At 8 months after administration of AAV1-LPL(S447X), an intravenous lipid challenge showed efficient, near-normal clearance of plasma TG. Histologic analyses of injected muscle further indicated that abnormal muscle morphology observed in LPL -/- mice was reversed after treatment. Expression of therapeutic levels of LPL(S447X), and the subsequent beneficial effect on plasma lipid levels, has lasted for more than 1 year. We therefore conclude that AAV1 mediated transfer of LPL(S447X) into murine skeletal muscle results in long-term near-correction of dyslipidemia associated with LPL deficiency. PMID- 15353047 TI - Near-patient testing in primary care. PMID- 15353048 TI - The journey towards patient-centredness. PMID- 15353049 TI - The course of C-reactive protein response in untreated upper respiratory tract infection. AB - BACKGROUND: High C-reactive protein (CRP) values are frequently found in patients with bacterial respiratory infection, and CRP testing has been shown to be useful in differentiating pneumonia from other respiratory infections. Raised CRP values may also be found in viral respiratory infection, and as a result there is a risk that antibiotics may be wrongly prescribed. AIMS: To describe the course of the CRP response during untreated upper respiratory tract infections and associations between the development of CRP values, erythrocyte sedimentation rate (ESR) and respiratory symptoms. DESIGN OF STUDY: Prospective study. SETTING: Seven general practices in northern Norway. METHOD: Patients with upper respiratory tract infection aged 16 years or over, who were not treated with antibiotics and who had been ill for no more than 3 days, were recruited. Microbiological examinations were undertaken, together with measurements of CRP, ESR and recording of symptoms daily during the first week of illness and on days 10, 14 and 21. RESULTS: An aetiological agent was established in 23 of the 41 included subjects. These were: influenza A, influenza B, rhinovirus, and other agents. Among the 15 patients examined on both the second and the third day of illness, the median CRP value increased from 7-10 mg/l, and the mean value was from 19-24 mg/l between day 2 and day 3. Peak CRP values were reached on days 2 to 4. Higher CRP values were found in those infected with influenza A and B than in the other subjects (P <0.001). A CRP value >10 mg/l was found in 26 subjects during the first 7 days, compared to five subjects after 1 week. Evidence of a secondary infection with group A streptococci was found in two of these five subjects. The development of the symptoms of sore throat, fatigue, clamminess, and pain from muscles and joints followed a similar course as the CRP response, while stuffy nose, cough, sputum production, and dyspnoea tended to persist after the CRP values had approached the normal range. CONCLUSION: A moderately elevated CRP value (10-60 mg/l) is a common finding in viral upper respiratory tract infection, with a peak during days 2-4 of illness. Moderately elevated CRP values cannot support a diagnosis of bacterial infection when the illness has lasted less than 7 days, but may indicate a complication of viral infection after a week. PMID- 15353050 TI - C-reactive protein measurement in general practice may lead to lower antibiotic prescribing for sinusitis. AB - BACKGROUND: Symptoms of bacterial sinusitis overlap with viral sinusitis, and it is difficult to distinguish between the two conditions based only on a clinical examination. Uncertain diagnosis results in the significant overuse of antibiotics, which is considered to be one of the most important reasons for development of bacterial resistance to antibiotics. A raised C-reactive protein (CRP) level is an indicator of bacterial infection and the CRP rapid test has been shown to be useful for the diagnosis of bacterial sinusitis in general practice. AIMS: To examine whether general practitioners (GPs) who use the CRP rapid test in their practice have a lower antibiotic prescribing rate for sinusitis than GPs who do not use the test. DESIGN OF STUDY: Observational design. SETTING: General practice in Denmark. METHOD: A group of GPs registered all contacts (n = 17 792) with patients who had respiratory tract infections during a 3-week period between 1 November 2001 and 31 January 2002. GPs who used a CRP rapid test were compared with GPs who did not, and the treatment of their patients (n = 1444) with suspected sinusitis was compared. RESULTS: A CRP rapid test was used by 77% (n = 281) of the GPs. In the group of GPs using a CRP rapid test, the rate of antibiotic prescribing was 59% (95% confidence interval [CI] = 56 to 62) compared with 78% (95% CI = 73 to 82) in the group of GPs who did not use a CRP test. Performing a CRP rapid test was the factor that exerted the greatest influence on whether the patients were prescribed antibiotics, and the level of CRP had a strong influence on the prescribing rate. CONCLUSION: The CRP rapid test has a substantial influence on the treatment of sinusitis, and implementing the test in general practice may lead to a reduction in antibiotic prescribing to patients with sinusitis. PMID- 15353051 TI - Eliciting patients' concerns: a randomised controlled trial of different approaches by the doctor. AB - BACKGROUND: Although a 'patient-centred' approach to general practice consultation is widely advocated, there is mixed evidence of its benefits. AIM: To measure the costs and benefits of using a prompt to elicit patients' concerns when they consult for minor illness. DESIGN OF STUDY: An open randomised controlled trial. SETTING: Four training semi-rural general practices in the south- east of the United Kingdom. METHOD: Patients identified during the first part of the consultation as having a self-limiting illness were randomised to a second part of the consultation that was conducted 'as usual' or involved a written prompt to elicit the patient's concerns. After each consultation the doctor noted the diagnosis and the consultation length and the patient self completed a questionnaire containing measures of satisfaction, enablement and anxiety. RESULTS: One hundred and ten patients were studied. Patients in the elicitation group reported a small but significant increase in the 'professional care' score of the consultation satisfaction questionnaire (88.2 versus 80.9, mean difference = 7.3, 95% confidence interval = 2.0 to 12.6) but no other benefits were detected. Consultations in the elicitation group, however, were longer by about a minute. CONCLUSION: Given the pressures on consultation time in general practice there must be questions about the practical value of eliciting patients' concerns if the benefit of doing so is small and the cost large. PMID- 15353052 TI - Giving patients an audiotape of their GP consultation: a randomised controlled trial. AB - BACKGROUND: Providing patients with an audiotape of their medical consultation has been a relatively common practice in oncology clinics for some years. However, broader generalisability of the technique has yet to be examined. AIMS: To investigate the efficacy of providing patients with an audiotape of their consultation in a general practice setting. DESIGN OF STUDY: Randomised controlled trial: 95 experimental participants, 85 controls. SETTING: Routine surgeries run by two general practitioners (GPs) in two different health centres. METHOD: All patients attending GP appointments were eligible for inclusion. Patients were followed up by telephone 7-10 days later. RESULTS: More than half (61%) of the patients who received a tape listened to it. Among listeners, 64% rated the tape useful or very useful; 24% noticed information not heard in the consultation. Half of listeners (46%) said that their understanding of the consultation improved after listening to the tape. Half of the listeners (48%) shared the tape with others, of whom 71% found sharing helpful or very helpful. However, 21% of those who shared the information with others found this unhelpful or very unhelpful, suggesting that patients may need to be briefed on the potential risks of sharing. At follow-up a week later, it emerged that being given a tape had no effect on adherence with GPs' advice, nor on anxiety about conditions. CONCLUSION: Providing patients with an audiotape of their GP consultation was positively rated by many patients. Although there were no detectable clinical effects at follow-up, the technique merits further evaluation in general practice. PMID- 15353053 TI - Influence of patient characteristics on doctors' questioning and lifestyle advice for coronary heart disease: a UK/US video experiment. AB - BACKGROUND: Risk factors for coronary heart disease (CHD) vary with patient characteristics but we do not know how this influences doctors' questioning and advice giving. AIMS: To find out whether four patient characteristics - age (55 versus 75 years), sex, class, and race - influence primary care doctors' questioning style and advice giving in the United Kingdom (UK) and United States (US). DESIGN OF STUDY: A factorial experiment using video simulation of a patient consulting with CHD symptoms, designed to systematically alter their age, sex, class, and race. SETTING: Surrey, south east London and the West Midlands in the UK, and Massachusetts in the US. METHOD: A stratified random sample of 128 general practitioners (GPs) in the UK and 128 primary care doctors in the US were shown video vignettes in their practices of patient consultations, and interviewed about patient management strategies. RESULTS: Sex and age influence doctors' questioning of patients presenting with CHD. Men are asked more questions overall, particularly about smoking and drinking. Middle-aged patients are asked more about their lifestyle. Advice about smoking is given to more men than women, and to more mid-life than older patients. Women doctors question patients about their lifestyle more often, and give more advice to patients about their diet. CONCLUSION: Doctors' questioning strategies are influenced by patients' sex and age, suggesting that doctors may miss smoking- and alcohol related factors among women and older patients with CHD. Doctors give more advice about smoking to men, despite sex equality in smoking prevalence. Therefore, doctors' information seeking and advice giving do not match known patient risk factors. PMID- 15353054 TI - Improving access to depression care: descriptive report of a multidisciplinary primary care pilot service. AB - BACKGROUND: Research has identified a need for improved depression care in primary care, while current United Kingdom (UK) health policy outlines standards for the management of the condition, including improved access to care. Innovative ways of working are needed to address these standards and provide better care. AIMS: To pilot a multidisciplinary service for the management of depressed patients with a particular focus on facilitating access. DESIGN OF STUDY: Uncontrolled descriptive pilot study. SETTING: One general practice in inner London. METHODS: The service was advertised by post to all 6689 adult patients registered with the practice. It provided open access and face-to-face assessment by a specially trained primary care nurse for patients who considered themselves to be depressed. Following assessment, depressed patients received systematic telephone support from nursing staff in addition to the usual care from the general practitioners (GPs). The ser vice was evaluated for a 6-month period. RESULTS: Sixty-six people, aged 19-77 years, 44 of them female, contacted the service, the majority in the first 2 months. Fifty-four patients were offered an assessment by the nurse. Thirty-five (80%) of the 44 attendees fulfilled criteria for major depression. Between them, the nurses and doctors achieved high levels of adherence to treatment and follow-up. This specialist service appears to have enabled a group of depressed patients, some of whom may not have sought or received help, to gain access to primary care. With appropriate supervision and training in depression care the nurses were able to assess and support depressed patients and this appeared to be acceptable to both patients and GPs. CONCLUSION: In its present form the service would not be cost-effective. However, we believe it could be adapted to suit the needs of individual or clusters of practices incorporating key elements of the service (open access and case management, in particular), and further evaluation by a controlled trial is suggested. PMID- 15353055 TI - Risk factors of influenza transmission in households. AB - BACKGROUND: Influenza transmission in households is a subject of renewed interest, as the vaccination of children is currently under debate and antiviral treatments have been approved for prophylactic use. AIMS: To quantify the risk factors of influenza transmission in households. DESIGN OF STUDY: A prospective study conducted during the 1999 to 2000 winter season in France. SETTING: Nine hundred and forty-six households where a member, the index patient, had visited their general practitioner (GP) because of an influenza-like illness were enrolled in the study. Five hundred and ten of the index patients tested positive for influenza A (subtype H3N2). A standardised daily questionnaire allowed for identification of secondary cases of influenza among their household contacts, who were followed-up for 15 days. Of the 395 (77%) households that completed the questionnaire, we selected 279 where no additional cases had occurred on the day of the index patient's visit to the GP. METHODS: Secondary cases of influenza were those household contacts who had developed clinical influenza within 5 days of the disease onset in the index patient. Hazard ratios for individual clinical and demographic characteristics of the contact and their index patient were derived from a Cox regression model. RESULTS: Overall in the 279 households, 131 (24.1%) secondary cases occurred among the 543 household contacts. There was an increased risk of influenza transmission in preschool contacts (hazard ratio [HR] = 1.85, 95% confidence interval [CI] = 1.09 to 3.26) as compared with school-age and adult contacts. There was also an increased risk in contacts exposed to preschool index patients (HR = 1.93, 95% CI = 1.09 to 3.42) and school-age index patients (HR = 1.68, 95% CI = 1.07 to 2.65), compared with those exposed to adult index cases. No other factor was associated with transmission of the disease. CONCLUSION: Our results support the major role of children in the dissemination of influenza in households. Vaccination of children or prophylaxis with neuraminidase inhibitors would prevent, respectively, 32-38% and 21-41% of secondary cases caused by exposure to a sick child in the household. PMID- 15353056 TI - Dependency rather than old age increases the risk of warfarin-related bleeding. AB - There is uncertainty about the safety of anticoagulation with warfarin in the elderly. This cohort study investigated the risk factors for warfarin-related bleeding in a primary care setting. In multiple regression analyses increased risk of bleeding was associated with domiciliary monitoring of international normalised ratio (INR), low proportion of INR tests in the target range, high intensity of anticoagulation, and male sex. Age was not associated with increased risk after adjustment. High levels of dependency, not old age, should raise concerns about the safety of anticoagulation with warfarin. PMID- 15353057 TI - Preference for a general practitioner and patients' evaluations of care: a cross sectional study. AB - Personal continuity is no longer always evident in general practice. Changes in society and in general practice seem to have shifted away from an emphasis on personal patient-doctor relationships. We studied how patients' lack of preference for a particular general practitioner (GP) or preference for a different GP is related to patients' evaluations of care. Patients who were indifferent to the GP seen, and patients who would have preferred another GP, evaluated consultations significantly less positively than patients who saw their GP of preference. Developments towards less personal doctoring in general practice should, therefore, be considered carefully. PMID- 15353058 TI - Cardiology. AB - This article describes recent developments in cardiology and cardiovascular disease that are likely to be relevant to primary healthcare professionals and their patients. The following subject areas are covered: Primary prevention: recent developments in pharmaco-logical interventions, drug interactions, and drugs that are likely to cause harm; cardiovascular risk estimation and shared decision making with patients; and new developments in 24-hour ambulatory blood pressure monitoring. Secondary prevention: new models of care, including nurse led care for the provision of hypertension and secondary prevention clinics; new drugs for the treatment of angina and myocardial infarction; ambulatory electrocardiography for the diagnosis of arrhythmias in primary care; and new developments in the treatment of atrial fibrillation - direct thrombin inhibitors and implantable devices. Tertiary prevention: recent developments in cardiac rehabilitation; recent evidence concerning revascularisation procedures and appropriateness criteria for referral; and implantable defibrillators. PMID- 15353059 TI - Are Scots with hypertension at high risk of diabetes or impaired glucose tolerance? PMID- 15353060 TI - Systematic review of service provisions to improve primary care access. PMID- 15353061 TI - To integrate or not to integrate? PMID- 15353062 TI - How would patients like to be addressed? A brief survey. PMID- 15353063 TI - Hepatitis B vaccination. PMID- 15353064 TI - Chlamydia screening in primary care. PMID- 15353065 TI - New concepts in screening. PMID- 15353066 TI - Advanced access. PMID- 15353068 TI - Showering with a green frog. PMID- 15353067 TI - A short walk! A feasible fitness test for general practice. PMID- 15353070 TI - The status of preimplantation genetic diagnosis in Japan: a criticism. AB - Advances in preimplantation genetic diagnosis (PGD) are occurring worldwide. New clinics specializing in this approach to the control of disease genes or imbalanced chromosome numbers in human preimplantation embryos continue to increase. One exception is Japan, where the Japanese Society of Obstetrics and Gynecology disapproves of this practice because it discriminates against people with genetic abnormalities. Yet, some doctors there wish to introduce this method to help their couples to improved forms of IVF. This paper stresses the rights of patients to have a healthy baby, if necessary by the use of PGD. It argues against prohibition, since it complements the current nature of prenatal diagnosis and avoids the need for abortions in case of afflicted embryos. Consideration is also given to other attempts at restriction that have failed. PMID- 15353071 TI - Case Report: grandmother, mother and another -- an intergenerational surrogacy using anonymous donated embryos. AB - Surrogacy and embryo donation remain beneficial but controversial infertility therapies. This is the first report of a pregnancy and subsequent birth of healthy twins following a surrogacy involving a female patient's biological mother as surrogate, but using anonymously donated embryos. Due to the novelty of combining intergenerational surrogacy with embryo donation, the legal side of adoption has taken over 3 years to complete. Intergenerational surrogacy using donated embryos can successfully provide a family with a third generation, offering an alternative to adoption of genetically unrelated children without involvement in the pregnancy. The legal and ethical aspects of the treatment and eventual adoption are discussed. PMID- 15353073 TI - Inhibition of nitric oxide synthesis potentiates apoptosis in the rabbit corpus luteum. AB - To determine if nitric oxide (NO) plays a role in corpus luteum (CL) physiology by affecting progesterone secretion or luteal apoptosis, an in-vitro pseudopregnant rabbit ovarian perfusion system was used to measure the effects of an inhibitor of NO synthesis, NG-nitro-L-arginine methyl ester (L-NAME), on progesterone secretion and corpus luteal apoptosis as measured by internucleosomal DNA breakdown. Pseudopregnant rabbit ovaries perfused in vitro with L-NAME did not demonstrate any significant differences compared with control ovaries in progesterone secretion. However, apoptosis, as measured by internucleosomal breakdown, was significantly increased in L-NAME-perfused CL compared with controls. While NO does not appear to directly affect progesterone secretion, there does appear to be a role for NO in CL maintenance, or a role for inhibition of NO production in CL regression. PMID- 15353074 TI - Enhanced oestradiol secretion briefly after embryo transfer in conception cycles from IVF. AB - The hypothesis was tested that conception cycles (CC) resulting from IVF can be distinguished from non-conception cycles (NC) by differences in corpora lutea function that are detectable at the earliest stage of embryo implantation. Luteal oestradiol secretion was analysed retrospectively in 409 ovarian stimulation cycles of 296 patients from the day of embryo transfer until 14 days after embryo transfer (ET+14) in IVF/intracytoplasmic sperm injection (ICSI) cycles. Human chorionic gonadotrophin (HCG) was administered in 330 of 409 cycles in addition to vaginal progesterone in all cycles. Differences in serum oestradiol concentrations between CC and NC increased from day ET+1 onward and became statistically significant on days ET+4 through ET+14, with higher oestradiol concentrations in CC compared with NC. Even though exogenous HCG administration prevented the fall in luteal oestradiol concentrations after ET+4 both in CC and NC, increasing differences in oestradiol concentrations between CC and NC after embryo transfer were observed in both groups of HCG-supplemented and non supplemented cycles. It is concluded that luteal oestradiol secretion is affected at the earliest stage of embryo implantation. The putative early signal to the corpus luteum associated with embryo attachment and early implantation appears to be superimposed onto the effect of exogenous luteal HCG administration and is clearly distinguishable as early as 4 days after embryo transfer in conception cycles. PMID- 15353077 TI - Oestradiol, cyclodextrin-encapsulated 17beta-oestradiol and the oestradiol solubilizer 2-hydroxypropyl-beta-cyclodextrin all impair preimplantation mouse embryo development. AB - The aim of this study was to examine the effects of 2-hydroxypropyl-beta cyclodextrin (HbetaC) used as a solubilizer for oestradiol, 17beta-oestradiol (ethanol soluble) and HbetaC-encapsulated-17beta-oestradiol on mouse embryo development in vitro. HbetaC had no effect on day 3 development. In contrast, blastocyst development and blastocyst cell number were significantly reduced in the presence of 10(-4) mol/l solubilizer equivalent, but not at lower concentrations. The proportion of compacted embryos was significantly reduced with 10(-4) mol/l 17beta-oestradiol. No blastocysts were formed at 10(-4) mol/l concentration of 17beta-oestradiol, although the rate of blastocyst formation did not differ at lower concentrations. Blastocyst cell number was significantly decreased compared with controls at 10(-5) mol/l 17beta-oestradiol. The dose response using HbetaC-encapsulated-17beta-oestradiol revealed that at 17beta oestradiol concentrations of 10(-4) and 10(-5) mol/l, blastocyst development was significantly reduced. Blastocyst cell number was significantly reduced compared with controls for all concentrations of HbetaC-encapsulated-17beta-oestradiol. Exposure of embryos to 17beta-oestradiol (10(-4) mol/l) reduced blastocyst development on days 4 and 5 significantly in cultures initiated at the zygote, 2 cell and 8-cell, but not the morulae, stages of development. Trophectoderm, ICM and blastocyst cell numbers as well as percentage ICM development were reduced significantly, regardless of the stage of development. Therefore, 17beta oestradiol does compromise embryo development. PMID- 15353079 TI - Human ovarian tissue cultures: extracellular matrix composition, coating density and tissue dimensions. AB - Human ovarian tissue can be successfully cryopreserved for fertility preservation. Optimal use of this approach requires the development of reliable restoration methods, including in-vitro culture of follicles. A culture system has been established, but improvement of the basic handling and techniques is necessary. Ovarian biopsies were collected from 33 women, cut into small pieces and cultured for 7-14 days on an extracellular matrix. Three separate studies investigated tissue dimensions (slices and cubes), coating density of extracellular matrix (diluted, thin and thick), and different extracellular matrix compositions (regular Matrigel, growth factor reduced Matrigel and laminin). Initial recruitment of primordial follicles and reduction in follicle viability was observed in all cultures compared with uncultured tissue. After 7 days of culture, more viable follicles were present in the cubed tissue, which also showed significant activation of growth, observed in tissue slices only after 14 days of culture. A diluted coating of Matrigel supported a greater proportion of viable follicles in 7-day cultures, whereas composition of the extracellular matrix had no effect. Human ovarian follicles can grow and develop in vitro within cortical tissue, and may benefit from culture as cubes on diluted Matrigel. This technique may provide a solution to the successful recovery and growth of follicles from frozen human ovarian tissue even though it will take time and much more optimization before it can be used in clinical practice. PMID- 15353080 TI - Role of INSL3 and LGR8 in cryptorchidism and testicular functions. AB - Cryptorchidism is the most frequent congenital anomaly of the urogenital tract in human males. INSL3 and LGR8/GREAT proteins seem to act as ligand and receptor respectively, and to have a role in gubernaculum development involved in testicular descent. Mutations in the INSL3 gene or LGR8/GREAT were found to be associated with cryptorchidism in humans. In a cohort of 135 ex-cryptorchid patients and 100 controls, mutations were sought in INSL3 and LGR8/GREAT genes by sequencing. Six patients were found with mutations in the INSL3 gene and four patients with LGR8/GREAT mutation (10/135, 7.4%). The 10 patients show different phenotypes, ranging from normozoospermia to complete azoospermia, and from bilateral cryptorchidism to retractile testes. Furthermore, the endocrine function of the testis appeared normal in all subjects. These findings demonstrate that INSL3-LGR8/GREAT mutations are frequently associated with human cryptorchidism, and that the only clinical consequence of alterations of the INSL3-LGR8/GREAT system seems to be failure of the testis to descend normally in the scrotum during embryonic development, without affecting the spermatogenic and endocrine components of the testis itself. The first analysis in humans of INSL3 was then performed using a novel radioimmunoassay kit to measure INSL3 concentrations in serum of adults. The results show that INSL3 circulates in adult men, it is a male-specific hormone, and it is of almost exclusively testicular origin. The role of this hormonal system in adulthood is, however, to date unknown. PMID- 15353081 TI - Role of utero-ovarian vascular impedance: predictor of ongoing pregnancy in an IVF-embryo transfer programme. AB - The objective of this study was to evaluate the role of uterine, endometrial and follicular blood flow in prediction of ongoing pregnancy after assisted conception. A prospective observational study was conducted on 53 women undergoing IVF treatment. Transvaginal colour and pulsed Doppler measurements were performed on the day that pituitary suppression was confirmed, on day 10 of ovarian stimulation and on the day prior to human chorionic gonadotrophin injection. On the last day of ovarian stimulation, blood flow in the ascending uterine artery of the women who would conceive was characterized by significantly lower pulsatility index values. Sub-endometrial vascular impedance was comparable in the pregnant and non-pregnant groups. There were no differences in the perifollicular vascularity between pregnant and non-pregnant women. The chance of achieving pregnancy predicted by uterine artery Doppler and perifollicular blood flow in women whose PI values were higher than 3.26 and 1.08 was very low, with a sensitivity of 1.00 and specificity of 0.59 and 0.82 respectively. The data provide evidence for an association between utero-ovarian perfusion and reproductive outcome following IVF treatment. Uterine and ovarian vascular impedance values identify those women whose pregnancy chances are significantly limited. Measures to decrease vascular impedance in such women might enhance pregnancy rates by improving embryo quality and uterine receptivity for implantation. PMID- 15353082 TI - Sex differences of abortuses and neonates in women with allo-immune recurrent abortions. AB - To investigate the possible association of recurrent spontaneous abortions (RSA) of immune aetiologies with neonatal sex differences, karyotyping of abortuses from allo-immune RSA and epidemiological studies on the sex differences of neonates from sporadic aborters was carried out. Allo-immune disorders, as diagnosed by an increased number of shared HLA class II loci and reduced blocking activity of the woman's serum in mixed lymphocyte reaction, were found almost twice (54.9%) as often as auto-immune disorders (29.9%) among a total of 244 women with RSA. Of 33 abortuses karyotyped from women with RSA, 69.7% showed normal female karyotypes, while only 6.1% had normal male karyotypes, indicating that female fetuses are more prone to abort than males. Epidemiological studies revealed that boys were born at a significantly greater incidence of 58.1% in 221 women with a history of sporadic abortion than 47.6% in 893 women with no history of abortion. Moreover, the proportion of women giving birth to boys only was consistently and significantly higher, regardless of repeated deliveries, in sporadic aborters (36.7%) than in women with no history of abortion (19.6%), showing that more boys were born than girls to women with sporadic abortion. It is concluded that male fetuses are more likely to survive than females in allo immune RSA due to allo-immune reproductive wastage of chromosomally normal female concept in early human pregnancy, and that allo-immune RSA makes up the highest proportion of unexplained RSA. PMID- 15353083 TI - Sperm aneuploidy and recurrent pregnancy loss. AB - Experiments of double target in-situ hybridization were performed separately for chromosomes 1-17, 8-18 and sex chromosomes on sperm samples from 20 couples suffering from three or more recurrent first trimester abortions. For a subset of this study population, additional experiments of multicolour fluorescence in-situ hybridization for chromosomes 4, 7, 12, 13, 15, 18, 21, and 22, were performed on the bases of the available data from abortive tissue karyotyping. A markedly high rate of sperm disomy (14.5-15.5%) was scored in only two cases. For three other patients, the cumulative disomy rates for chromosomes 1, 17, 8, 18, X and Y also increased but at a lower level (7.8-9.5%). For the remaining 15 patients, the frequency of sperm aneuploidy was moderately increased or normal. Men with recurrent pregnancy loss (RPL) and poor semen quality had baseline sperm aneuploidy and diploidy rates higher than men with normal semen parameters (with or without RPL). Using probes for chromosomes 1, 17, 8, 18, X and Y, significantly elevated frequencies of sperm aneuploidy (not diploidy) were found in 10% of men with a history of RPL. Their rate of sperm aneuploidy was 30-34%. For the other men, changes in sperm aneuploidy were not thought to affect RPL. Poor semen quality per se impacted negatively on sperm aneuploidy and diploidy, thus making the interpretation of clinical data more difficult. PMID- 15353084 TI - Nine-day-old human embryo cultured in vitro: a clue to the origins of embryonic stem cells. AB - The aims of this study were to investigate whether the human embryo could sustain development beyond the blastocyst stage in vitro and to identify the precise origins of embryonic stem cells (ES cells) from the embryoblast. A frozen-thawed 4-cell embryo was cultured to the post-blastocyst stage. This 9-day-old embryo presented a solid mass of inner cells (resembling a tumour) surrounded by surface trophoblast cells. Clumps of multinucleated syncytiotrophoblast cells were evident at one pole. Most cells resembled those of blastocysts. However, there were groups of comparatively undifferentiated cells within the inner cell mass somewhat resembling ES cells documented previously, that might give a clue as to their origins. The embryo attempted to form an amnion with a cavity, but did not present a bilaminar, discoidal structure as expected in week 2 of development, and hence was abnormal. PMID- 15353085 TI - Human blastocysts for the development of embryonic stem cells. AB - Establishment of human embryonic stem cells (hES) from surplus human IVF embryos has been successful when both fresh and frozen-thawed cleavage stage embryos have been cultured to the blastocyst stage. This study reports the characteristics of the starting material, the blastocysts, for hES cell lines that were first derived at the University of Gothenburg, Sahlgrenska University Hospital in 1999. Twenty-two hES cell lines were derived by Cellartis AB from 114 blastocysts, giving an overall success rate of 19.3%. The blastocysts from which the hES cell lines were established were of varying morphological quality, both fresh and frozen-thawed. Two techniques of hES establishment were applied, i.e. direct application of the blastocysts on feeder cells or the standard immunosurgery method. It was further found that the efficiency by which frozen-thawed embryos gave rise to new hES cell lines was 3.7 times better than with fresh surplus embryos. These findings suggest that frozen-thawed embryos are superior to fresh surplus human embryos in hES cell establishment, which also avoids specific ethical problems associated with embryo donation in a fresh IVF cycle. PMID- 15353087 TI - Role of free radicals in female reproductive diseases and assisted reproduction. AB - Infertility is a common problem experienced by many couples. Numerous treatments are available for female infertility. However, in some cases, the treatment is empirical in nature because the aetiology of infertility is not fully understood. Recently, reactive oxygen species (ROS) have been shown to have an important role in the normal functioning of reproductive system and in the pathogenesis of infertility in females. Reactive oxygen species may also play a role in other reproductive organ diseases of women such as endometriosis. Oxidative stress develops when there is an imbalance between the generation of ROS and the scavenging capacity of antioxidants in the reproductive tract. It affects both natural and assisted fertility. Because assisted reproductive techniques are used extensively in the treatment of infertility, it is critical to understand the in vitro conditions that affect fertilization and embryo development. Treatments that reduce oxidative stress may help infertile women with diseases that are caused by this imbalance. Such strategies include identifying the source of excessive generation of ROS, treating the primary cause, and in-vitro and in-vivo supplementation of antioxidants. Research is in progress to identify the mechanisms that are involved in the aetiology of female reproductive diseases caused by ROS, and to create effective strategies that can counteract oxidative stress. PMID- 15353086 TI - Cultures of human embryonic stem cells: serum replacement medium or serum containing media and the effect of basic fibroblast growth factor. AB - Human embryonic stem (hES) cells have traditionally been cultured in medium containing fetal calf serum (FCS) and mouse fibroblasts as feeder cells. The use of animal derived materials carries a risk of transmitting animal pathogens, and they are not optimal in cultures aimed at cell transplantation in humans. This technical study aiming at facilitating IVF units to establish new hES cell lines, has systematically compared the non-differentiated growth of the hES cell line HS237, originally derived and thereafter cultured using human foreskin fibroblasts as feeder cells, by culturing it in media containing serum replacement (SR; 10, 15, 20%), FCS, and human serum. In addition, optimal concentrations of insulin-transferrin-selenium (ITS) mixture and the effect of basic fibroblast growth factor (bFGF) have also been studied. Cellular growth was monitored daily and maintenance of their non-differentiated character was studied using antibodies against TRA-1-60, TRA-1-81 and SSEA-4 and expression of Oct-4. The hES cells proliferated fastest when 20% of SR was used. In human serum containing medium, the cells underwent extensive spontaneous differentiation within a few passages. The FCS supported the non-differentiated growth poorly. Basic fibroblast growth factor supported non-differentiated growth, the highest concentration (8 ng/ml) giving the best result, while ITS was not beneficial. PMID- 15353089 TI - Should ICSI be used in non-male factor infertility? AB - There is general agreement that intracytoplasmic sperm injection (ICSI) should be used in male factor infertility cases, such as oligoasthenoteratozoospermia, presence of anti-sperm antibodies, or azoospermia, these cases being diagnosed through abnormal semen analysis. There are no randomized clinical trials comparing ICSI with IVF (or other interventions) where semen quality is so poor that IVF would not achieve fertilization. It is accepted that ICSI is the only treatment option in those circumstances. The role of ICSI where IVF can be expected to give a reasonable fertilization rate is the question that needs to be answered. The argument is whether or not ICSI should be used for all cases of infertility. This paper proposes and strongly supports the use of ICSI for all indications. Considerations of fertilization and embryo development, cost effectiveness and safety will be clearly discussed. PMID- 15353094 TI - A survey of complementary therapy services provided by hospices. AB - A questionnaire was mailed to 300 randomly selected hospices in the United States, to gather preliminary data on the nature of complementary therapy services provided by hospices. Information included types of complementary therapies offered, utilization, staffing, obstacles, as well as suggestions for improving hospice complementary therapy services. Of a total of 169 responding hospices, 60% offered complementary therapies to patients. The most popular therapies were massage therapy and music therapy. Only a portion of patients in these hospices received complementary therapy. Many hospices were limited in the amount of complementary therapy services they could provide because of program constraints, such as funding problems, lack of qualified complementary staff, inadequate knowledge of complementary therapies and how to offer these services, and resistance to complementary therapies by some staff and patients. A crucial challenge for hospices interested in providing complementary therapies to patients is to find ways to overcome these obstacles. PMID- 15353095 TI - Ethical wills and suffering in patients with cancer: a pilot study. AB - INTRODUCTION: Suffering at the end of life may be caused by many factors, including pain and other symptoms, concern about family and friends, and loss of control of one's life. Several authors have suggested that loss of meaning is pivotal in suffering. An ethical will (EW) is a statement, usually written, capturing one's values, wisdom, hopes, and advice. EWs have been suggested as a vehicle for finding meaning as the end of life approaches. This pilot study of EWs examined methods for exploring the role of EWs in reducing suffering at the end of life. METHODS: Oncology clinic patients 65+ years of age in active therapy for cancer were randomly assigned to one of two arms: EW or control. Subjects in both arms had writing assignments, three home visits, and exit interviews. Suffering was measured at baseline and at the time of the exit interview, using a series of Likert-like scales. RESULTS: Twenty-four subjects (10 EW and 14 control) completed the study. Among EW subjects, trends toward reduced suffering were noted in "concern for loved ones," "unfinished business," and "fear of the future." Several methodological issues were identified by this pilot study, including selection of population for studying suffering, placebo effect, and randomization. CONCLUSIONS: EW may be valuable in alleviating suffering; a larger study will be needed to examine efficacy. The study of interventions designed to reduce suffering at the end of life requires careful attention to the definition and measurement of suffering, study design, and subject selection. PMID- 15353096 TI - Are depressed patients more likely to share health care decisions with others? AB - OBJECTIVES: To determine whether psychological variables, particularly depression, influence patients' willingness to share medical decisions with family members or friends. DESIGN: Cross-sectional interviews. SETTING: Oncology and general geriatrics outpatient clinics of an urban VA medical center. PARTICIPANTS: Ninety-five patients with a Charlson comorbidity index score greater than 5. MEASUREMENTS: Subjects described the way that they make health care decisions with friends or family as a dichotomous variable ("shared" versus "not shared") and as a 5-point ordinal variable (the degree to which they share decisions). Patients also completed the 15-item version of the Geriatric Depression Scale (GDS), the Global Distress Index of the Memorial Symptom Assessment Scale, and selected tests of cognitive function and health literacy. RESULTS: Patients with a GDS score higher than 5 were more likely to share decision-making (16/26 versus 26/69; odds ratio 2.58; p = 0.040), as were patients who were married (23/35 versus 19/60; odds ratio 3.63; p = 0.001). In multivariable regression models, a short form GDS score higher than 5 was independently associated with a willingness to share decision with others. CONCLUSION: These results suggest that depression may have a clinically significant influence on patients' willingness to share health care decisions with others. Health care providers should be alert to this possibility, particularly when the decision at hand is significant. PMID- 15353097 TI - "Getting everyone on the same page": nursing home physicians' perspectives on end of-life care. AB - PURPOSE: To improve understanding of nursing home physicians' perspectives regarding end-of-life care, and to suggest directions for further research. METHODS: An exploratory qualitative design based on interviews of 12 nursing home physicians, 10 of whom were medical directors. Medical students served as interviewers. SAMPLE: A purposeful sampling strategy yielded interviews with 12 physicians. The sample was selected based on "intensity sampling," which seeks information-rich but not extreme cases. Ten of the 12 physicians were nursing home medical directors; all respondents practiced at least 4 years part-time or full-time in a nursing home setting. Respondents varied by age, gender, urban/rural location, and fellowship training (half the sample had completed a geriatrics fellowship). Seven physicians were affiliated with an academic medical center. RESULTS: Four themes were identified in the analysis of the 12 interview transcripts: extensive familiarity with dying; consensus is integral to good end of-life care; obstacles can interfere with consensus; and advance directives set the stage for conversations about end-of-life care. The importance of consensus, both in terms of prognosis and in developing a palliative care plan, emerged as the major finding. CONCLUSIONS: For the 12 physicians in this study consensus about the resident's status and an appropriate care plan are important features of good end-of-life care. Further research is needed to determine if other members of the health care team (i.e., residents, family members, nursing staff, social worker, etc.) also value consensus highly. It will be important to determine what barriers to consensus other team members identify. Based on the understanding generated from this study, a refinement of the general Education for Physicians on End-of-Life Care (EPEC) model describing the relationship between curative and palliative care is proposed for nursing homes. The refinement underscores the points at which the team might consider revisiting consensus about the resident's status and care plan. PMID- 15353098 TI - Hydromorphone neuroexcitation. AB - Continuous parenteral hydromorphone is used to treat pain in palliative care. Case reports have suggested that neuroexcitatory symptoms, such as agitation, myoclonic activity, and even seizures may occur during administration. However, little information exists on the incidence of these side effects or their relationship to the dose or duration of parenteral hydromorphone. A retrospective chart review was performed on 48 terminally ill hospice patients who received continuous parenteral hydromorphone for pain control. Chart reviews were conducted searching for three neuroexcitatory symptoms: agitation, myoclonus, and seizures; the incidence and relationship of these symptoms were statistically compared to the maximal dose and number of days on continuous parenteral hydromorphone. We found that agitation, myoclonus, and seizures were not associated with the patients gender, age, or diagnosis but found that agitation was associated (p < 0.01) in patients with known metastatic disease. Agitation, myoclonus, and seizures were independently associated with the maximal dose (p < 0.05, p < 0.001, and p < 0.05) and with the duration (p < 0.01, p < 0.05, and p < 0.01) of continuous parenteral hydromorphone A possible mechanism for these findings is hydromorphone-3-glucoronide, a metabolic product of hydromorphone, which has been implicated in neuroexcitatory symptoms in laboratory investigations. PMID- 15353099 TI - Individual quality of life is not correlated with health-related quality of life or physical function in patients with amyotrophic lateral sclerosis. AB - We compared the change over time of individual quality of life (QOL) versus health-related QOL (HRQOL) and functional status in palliative care patients with amyotrophic lateral sclerosis (ALS). Forty-two patients with ALS performed the following assessments: the ALS functional rating scale (ALSFRS), the Sickness Impact Profile (SIP), the Short Form 36 (SF-36), and the Schedule for the Evaluation of Individual QOL-Direct Weighting (SEIQOL-DW). The SF-36 and the SEIQOL-DW were assigned at random. Patients were examined at least three times at 2-month intervals. There was a significant decrease from visit 1 to 3 (4-month period) in the function-based values of the SIP, SF-36, and ALSFRS. Despite this progressive decline of physical function and HRQOL, individual QOL as assessed by the SEIQOL-DW remained stable throughout the observation period. Correspondingly, there was a clear correlation between the ALSFRS, the SIP, and the SF-36, but no correlation between these scales and the SEIQOL-DW. The QOL domains most often named in the SEIQOL-DW were family, friends/social life, health, and profession. Thus, individual QOL appears to be largely independent from physical function in severely ill patients with ALS. Because of the characteristics of the scale, assessment of individual QOL may have an interventional value in palliative care. PMID- 15353100 TI - Jewish medical ethics and end-of-life care. AB - While Judaism espouses the infinite value of human life, Judaism recognizes that all life is finite and, as such, its teachings are compatible with the principles of palliative medicine and end-of-life care as they are currently practiced. Jewish medical ethics as derived from Jewish law, has definitions for the four cardinal values of secular medical ethics: autonomy, beneficence, nonmaleficence, and justice, with the major difference between Jewish law and secular medical ethics being that orthodox or traditional Jews are perceived to limit their autonomy by choosing, with the assistance and advice of their rabbis, to follow God's law as defined by the Bible and post-Biblical sources. With an understanding of Jewish medical ethics as defined by Jewish law, various issues pertaining to the care of Jewish patients who are near the end-of-life can be better understood. Jewish tradition contains within its textual sources the concept of terminal illness. The shortening of life through suicide, assisted suicide, or euthanasia is categorically forbidden. For patients who are terminally ill, treatments that are not potentially curative may be refused, especially when harm may result. Under certain circumstances, treatments may be withheld, but active treatment already started may not usually be withdrawn. While patients should generally not be lied to regarding their conditions, withholding information or even providing false information may be appropriate when it is felt that the truth will cause significant harm. Pain and suffering must be treated aggressively, even if there is an indirect risk of unintentionally shortening life. Finally, patients may execute advance directives, providing that the patient's rabbi is involved in the process. PMID- 15353101 TI - Update in hospice and palliative care 2003. PMID- 15353102 TI - Neuroexcitatory effects of opioids: patient assessment #57. PMID- 15353103 TI - Neuroexcitatory effects of opioids: treatment #58. PMID- 15353109 TI - JPM patient information. Care of the dying patient at home. PMID- 15353105 TI - Care of the Christian Science patient. PMID- 15353111 TI - Causes and consequences of fever complicating critical surgical illness. AB - BACKGROUND: Fever may have malign consequences in the postoperative period. This study was performed to determine the causes and consequences of fever in critically ill surgical patients. The specific hypothesis tested is that postoperative fever is associated with adverse clinical outcomes, including increased organ dysfunction and risk of death. METHODS: Inception-cohort study of critically ill surgical patients who manifested a core temperature of >/=38.2 degrees C for the first time. The episode of fever was monitored until resolution, which was defined as a core temperature of <38.2 degrees C for at least 72 consecutive h. Demographic data collected included age, gender, admission diagnosis, admission status (elective/emergency), severity of illness (APACHE III), the systemic inflammatory response syndrome (SIRS) score, the cumulative multiple organ dysfunction score, cause of fever (infectious/non infectious), ICU and hospital length of stay, and mortality. The day of onset of fever in the ICU, peak temperature, ICU day of peak temperature, and duration of fever episode were recorded. All diagnostic and therapeutic interventions were recorded, including the type and duration of antibiotic therapy. Univariate results of possible significance (alpha < 0.15) were tested in logistic regression models for independence of effect upon mortality after auto correlation was excluded by matrix correlations and the Durbin-Watson statistic. Cases where both non-infectious and infectious causes of fever were present were analyzed as part of the infectious group, whereas the cumulative MOD score was dichotomized (< 5, >/=5 points) at a value known to be associated with increased mortality. RESULTS: Among 2,419 screened patients, 626 patients (26%) developed fever. Febrile patients were older, sicker, more likely to have undergone emergency surgery, more likely to develop organ dysfunction, and more likely to die (all, p < 0.0001). The mean day of onset of fever was day 1 and the mean peak temperature for the episode was 39.1 +/- 0.1 degrees C. For most patients, it was their only episode of fever, with a mean of 1.4 +/- 0.1 episodes/patient. Forty six percent of febrile patients were found to have an infectious cause of fever. Nearly all patients had SIRS, and nearly all developed organ dysfunction to some degree. By logistic regression, the presence of SIRS (as opposed to fever in isolation), emergency status, higher APACHE III score and the peak temperature were associated with increased mortality, with peak temperature being the most powerful predictor in the model (OR 2.20, 95% Cl 1.57-3.19). Gender had no bearing on outcome, and there was a trend toward a protective effect from an infectious etiology of fever. CONCLUSIONS: Postoperative fever is deleterious to critically ill patients. The magnitude of fever is a determinant of mortality, whereas an infectious etiology of fever may not be. The impacts of nosocomial infection and suppression of fever on critically surgical patients deserve further study. PMID- 15353112 TI - The burden of appendicitis-related hospitalizations in the United States in 1997. AB - BACKGROUND: Acute appendicitis is among the most frequent causes of surgical abdominal disease worldwide. METHODS: Data from the Nationwide Inpatient Sample of the Healthcare Utilization Project were used to estimate the prevalence and disease burden of appendicitis-related hospitalizations in the United States in 1997. The data are a representative sample of discharge records from community hospitals drawn from 22 states in the United States. RESULTS: In the United States in 1997, there were an estimated 252,682 (95% CI: 242,957-262,407) appendicitis-related hospitalizations. The mean length of stay and total charges for appendicitis-related hospitalizations were four days (95% CI: 3.4-4.6) and $11,645 (95% CI: $11,299-$11,992) per hospitalization, respectively. Appendicitis related hospitalizations associated with post-operative infection, peritoneal abscess, or peritonitis had longer average lengths of stay and higher average costs when compared to hospitalizations associated with local appendicitis without post-operative infection. Appendicitis cases among very young and older patients were more likely to be associated with peritoneal abscess, peritonitis, or post-operative infection. CONCLUSIONS: In 1997, appendicitis-related hospitalizations accounted for 0.6% of all hospitalizations in the United States, resulting in approximately one million hospital days and $3 billion in hospital charges. Between 1984 and 1997, the rate of appendicitis hospitalizations in the U. S. population declined slightly, whereas the total number of hospital days remained unchanged. PMID- 15353113 TI - Pulmonary MnSOD is nitrated following hepatic ischemia-reperfusion. AB - BACKGROUND: Ischemia-reperfusion (I/R) of remote organs is a common cause of lung injury. We observed that lung injury after partial hepatic I/R in mice coincides with the appearance of 3-nitrotyrosine (NT) in the lung tissue, a marker of peroxynitrite involvement and oxidant stress. Peroxynitrite can cause mitochondrial dysfunction by inactivation of manganese superoxide dismutase (MnSOD), the major antioxidant enzyme in mitochondria. Our aims were to examine whether pulmonary MnSOD is a target of nitration following hepatic I/R and whether nitrated MnSOD (N-MnSOD) correlates with acute lung injury. METHODS: Five 20-25-g male C57BL/6 mice underwent laparotomy, and atraumatic occlusion of the portal and arterial blood supply to the upper three lobes of the liver for 90 min. This warm ischemic period was followed by 4 h of reperfusion, and the animals were then euthanized. Lung injury was assessed by LDH and protein levels in bronchoalveolar lavage (BAL) fluid. Pulmonary MnSOD activity in pulmonary homogenates was measured by the cytochrome c reduction method. The presence of N MnSOD was determined by immunoprecipitation (IP) and Western Blot analysis. Controls (N = 5) underwent sham operation. RESULTS: Elevated plasma transaminases confirmed hepatic injury. Lung injury was demonstrated by elevation in BAL protein and LDH levels (495.7 (48.4) versus 644.9 (37.3) [p < 0.05] and 56.5 (11.8) versus 345.2 (80) [p < 0.01], respectively). Immunoprecipitation and Western blot demonstrated N-MnSOD in the lung tissue of I/R animals but not controls. MnSOD activity decreased following I/R (8.1 (0.7) versus 10.8 (0.3) [p < 0.05]). CONCLUSIONS: Pulmonary MnSOD is both nitrated and inactivated following hepatic I/R and is associated with acute lung injury. These findings suggest that MnSOD incapacitance may contribute to I/R-induced lung injury and provide a therapeutic target in attenuating multisystem injury following hepatic I/R. PMID- 15353114 TI - Local infections after above-knee prosthetic femoropopliteal bypass for intermittent claudication. AB - BACKGROUND: The use of prosthetic grafts in the treatment of intermittent claudication is still a controversy. Prosthetic bypass for this usually benign condition may in some cases lead to a graft infection. This potentially disastrous complication is difficult to manage. METHODS: One hundred forty-one consecutive operations done on 129 patients between January, 1990 and December, 2001 in a single university vascular unit have been studied. Lymphatic complications and local infections have been related to preoperative risk factors, complications, secondary procedures, and outcome. RESULTS: During a mean follow-up period of 42 months, lymph complications occurred after 18 operations, surgical site infection after 11 procedures, and graft infection after 17 operations. Eleven infected grafts were treated successfully without graft excision. Six of these grafts healed with antibiotics only. The risk of developing a local infection was significantly correlated with postoperative lymph fistula. Reoperative surgery was associated with graft infection. Graft infection caused by Staphylococcus aureus always warranted surgery, either local revision or graft excision. CONCLUSIONS: The present series had a high frequency of graft infections. Our data suggest that a selective approach should be taken towards excision of infected femoropopliteal prostheses. The need for and extent of surgery should be individualized according to the clinical presentation of the graft infection and the type of bacteria involved. We advocate a conservative attitude towards surgical treatment of intermittent claudication. PMID- 15353115 TI - Methicillin-resistant Staphylococcus aureus infections in vascular surgery: increasing prevalence. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is a virulent organism that causes substantial infection-related morbidity and mortality in hospitalized patients. For example, MRSA infection of prosthetic vascular grafts can be limb- and life-threatening in surgical patients. We investigated the incidence of MRSA infection in vascular surgical patients who developed postoperative infectious complications. METHODS: Prospective data (including procedure, nature of complication, and outcome) were collected on all vascular surgery patients (n = 772) over a two-year period from January, 2000, to December, 2001. The study cohort included all patients who developed postoperative complications (n = 119, 15.4%), including infection, unplanned or prolonged intubation, metabolic abnormalities, and myocardial ischemia. Bacteriologic data of all clinical cultures were reviewed and infection-related outcomes were examined. RESULTS: Hospital-acquired infection occurred in 79 (66.4%) of 119 patients who developed postoperative complications. Review of microbiology data (total, 197 bacterial isolates) confirmed that gram-positive organisms were the leading cause of infections, with 120 gram-positive isolates (60.9%). Of the gram-positive isolates, S. aureus was the most common pathogen, isolated in 73 (60.8%) cultures. The single most prevalent organism was MRSA, identified in 42 of 73 (57.5%) of cultures. The site of MRSA infection was confirmed to be the surgical site (54.8%), sputum (17.0%), blood (14.3%), urine (9.5%), peritoneal fluid (2.4%), or catheter insertion site (2.4%). Only four patients had prior MRSA infection or colonization. Prosthetic vascular grafts were placed in 51 (42.9%) of the 119 vascular patients, and 26 patients (51.6%) acquired MRSA infection. Graft removal was required in three patients due to infection (MRSA infection in two patients, Pseudomonas aeruginosa in one patient). Forty percent of patients with MRSA infection required an amputation as their surgical procedure. No patient died of MRSA bacteremia during the study period. Mean hospital length of stay (LOS) was longer in patients with MRSA infection (29.6 d vs. 22.7 days, range 2-174 days, p < 0.05) compared to the total study cohort of vascular patients with postoperative infections. CONCLUSIONS: Methicillin-resistant S. aureus has emerged as the leading cause of postoperative infection in vascular surgery patients, and is associated with substantial morbidity, increased hospital LOS, and higher incidences of amputation and graft removal. Greater emphasis on preoperative screening protocols for MRSA colonization is warranted, in conjunction with aggressive infection control measures, alteration of preoperative prophylactic antimicrobial use in MRSA-colonized patients, and meticulous postoperative surveillance for MRSA infection. Furthermore, antimicrobial treatment of postoperative infectious complications in vascular surgery patients should include empiric coverage for MRSA in institutions where MRSA is endemic. PMID- 15353116 TI - Pyogenic liver abscess complicating biliary stricture due to chronic pancreatitis. AB - BACKGROUND: Chronic pancreatitis uncommonly causes common bile duct stricture, and common bile duct stricture rarely leads to pyogenic liver abscess. METHODS: We describe a 51-year-old man who developed a pyogenic liver abscess after the development of a common bile duct stricture due to chronic pancreatitis. RESULTS: The patient required open drainage of the liver abscess and decompressive choledochoduodenostomy. His infection was persistent and he required percutaneous and subsequent open drainage of a second hepatic abscess. The patient died of refractory sepsis and multi-organ failure. CONCLUSIONS: The relevant English literature concerning pyogenic liver abscesses, specifically in the setting of common bile duct stricture due to chronic pancreatitis, is reviewed and discussed. PMID- 15353117 TI - Perforated appendicitis in the elderly masquerading as acute coronary syndrome. AB - BACKGROUND: Evaluation of acute appendicitis in the elderly presents a diagnostic challenge to the surgeon. The diagnosis is often difficult to establish on account of general poverty of history and clinical signs, comorbidity and the unreliability of diagnostic procedures. Due to this, morbidity and mortality statistics in this group of patients is unacceptably higher than the general population. METHODS: Individual case report and literature review. RESULTS: We present an unusual case of perforated appendicitis masquerading as acute coronary syndrome. CONCLUSIONS: Patient's education, high index of suspicion, and an aggressive surgical approach is needed if the outcome among the elderly population is to be improved. PMID- 15353118 TI - Intramedullary abscess from group F Streptococcus. AB - BACKGROUND: Intramedullary abscesses are rare and may be misdiagnosed. We report the case of a 54-year-old man with carcinoma of the gastroesophageal junction who presented after thoracotomy with hemiparesis and a ring-enhancing intramedullary lesion at C2-3 on magnetic resonance imaging. METHODS: Individual case report and literature review. RESULTS: Although the lesion was initially mistaken for metastasis, at surgery an intramedullary abscess was discovered, from which group F Streptococcus was isolated. Conclusions : Although other streptococcal species have been found in abscesses of the spinal cord, this particular strain has not been identified previously as a cause of such abscesses. Prompt surgical evacuation is recommended, and should be followed by appropriately tailored antibiotic therapy. PMID- 15353119 TI - Fulminant liver failure following infection by Clostridium perfringens. AB - BACKGROUND: Since the first description of gas gangrene of an internal organ by Fraenkel in 1889, few cases of acute organ failure following Clostridium perfringens infection have been described in the medical literature. Isolated Clostridium perfringens infection with subsequent sepsis syndrome is an extremely rare clinical syndrome. A consecutive pattern of multiple organ failure generally has a very high mortality rate. METHODS: Individual case report and literature review. RESULTS: A 58-year-old male patient developed fulminant necrotic liver failure following a Clostridium perfringens infection. Despite all intensive care measures, including computed tomography-guided drainage, the condition of the patient deteriorated rapidly and the patient died. In this case report, we characterize the symptoms of gas gangrene isolated to the liver and compare the treatment measures instituted with the medical literature. CONCLUSIONS: In our presented case, primary malignant disease of the papilla of Vater and resection by a Whipple procedure with a hepatico-jejunostomy were a decisive cause of the gas gangrene in the liver. The origin is probably ascension up the common hepatic duct of gut-derived bacteria. PMID- 15353120 TI - Hand infections: basic principles and management. AB - BACKGROUND: An untreated hand infection can lead rapidly to tissue destruction and permanent disability. METHODS: Review of the pertinent English literature. RESULTS: Early diagnosis and timely, effective intervention is crucial to treatment. Anatomically, the hand consists of multiple potential spaces and tissue planes that can become infected with a wide variety of organisms. Thus, a working knowledge of hand anatomy as well as pharmalogical therapies is imperative for successful management. CONCLUSIONS: Knowledge of the basic principles of hand care, relevant anatomy, infectious organisms, and clinical signs of an unusually aggressive infection greatly improves treatment efficacy. PMID- 15353122 TI - Outcomes/cost effectiveness and clinical suspicion of appendicitis. PMID- 15353123 TI - Uterine receptivity and the plasma membrane transformation. AB - This review begins with a brief commentary on the diversity of placentation mechanisms, and then goes on to examine the extensive alterations which occur in the plasma membrane of uterine epithelial cells during early pregnancy across species. Ultrastructural, biochemical and more general morphological data reveal that strikingly common phenomena occur in this plasma membrane during early pregnancy despite the diversity of placental types--from epitheliochorial to hemochorial, which ultimately form in different species. To encapsulate the concept that common morphological and molecular alterations occur across species, that they are found basolaterally as well as apically, and that moreover they are an ongoing process during much of early pregnancy, not just an event at the time attachment, the term 'plasma membrane transformation' is suggested which also emphasises that alterations in this plasma membrane during early pregnancy are key to uterine receptivity. PMID- 15353124 TI - Identification of a novel population of human cord blood cells with hema topoietic and chondrocytic potential. AB - With the exception of mature erythrocytes, cells within the human hematopoietic system are characterized by the cell surface expression of the pan-leukocyte receptor CD45. Here, we identify a novel subset among mononuclear cord blood cells depleted of lineage commitment markers (Lin-) that are devoid of CD45 expression. Surprisingly, functional examination of Lin-CD45- cells also lacking cell surface CD34 revealed they were capable of multipotential hematopoietic progenitor capacity. Co-culture with mouse embryonic limb bud cells demonstrated that Lin-CD45-CD34- cells were capable of contributing to cartilage nodules and differentiating into human chondrocytes. BMP-4, a mesodermal factor known to promote chondrogenesis, significantly augmented Lin-CD45-CD34- differentiation into chondrocytes. Moreover, unlike CD34+ human hematopoietic stem cells, Lin CD45-CD34- cells were unable to proliferate or survive in liquid cultures, whereas single Lin-CD45-CD34- cells were able to chimerize the inner cell mass (ICM) of murine blastocysts and proliferate in this embryonic environment. Our study identifies a novel population of Lin-CD45-CD34- cells capable of commitment into both hematopoietic and chondrocytic lineages, suggesting that human cord blood may provide a more ubiquitous source of tissue with broader developmental potential than previously appreciated. PMID- 15353125 TI - A novel protein-DNA interaction involved with the CpG dinucleotide at -30 upstream is linked to the DNA methylation mediated transcription silencing of the MAGE-A1 gene. AB - To understand the DNA-methylation mediated gene silencing mechanisms, we analyzed in cell culture of the promoter function of the MAGE-A1 gene, which is frequently demethylated and over-expressed in human hepatocellular carcinoma. We have established the correlation of the DNA methylation of the promoter CpG island with expression status of this gene in a panel of the established liver cancer cell lines. The crucial CpG dinucleotide(s) within the minimal promoter subjected to the control mediated by DNA methylation with profound biological functions was also delineated. Furthermore, a novel sequence-specific DNA-protein interaction at the -30 CpG dinucleotide upstream of the gene was found having a vital part to play in the DNA methylation mediated transcription silencing of the MAGE-A1 gene. Our results would not only provide new insights into the DNA methylation mediated mechanisms over transcription of the MAGE-A1 gene, but also pave the way for further defining the cross-talk among DNA methylation, histone modification and chromatin remodeling in detail. PMID- 15353126 TI - The modulation of radiation-induced cell death by genistein in K562 cells: activation of thymidine kinase 1. AB - Ionizing radiation is one of the most effective tools in cancer therapy. In a previous study, we reported that protein tyrosine kinase (PTK) inhibitors modulate the radiation responses in the human chronic myelogenous leukemia (CML) cell line K562. The receptor tyrosine kinase inhibitor, genistein, delayed radiation-induced cell death, while non-recepter tyrosine kinase inhibitor, herbimycin A (HMA) enhances radiation-induced apoptosis. In this study, we focused on the modulation of radiation-induced cell death by genistein and performed PCR-select suppression subtractive hybridization (SSH) to understand its molecular mechanism. We identified human thymidine kinase 1 (TK1), which is cell cycle regulatory gene and confirmed expression of TK1 mRNA by Northern blot analysis. Expression of TK1 mRNA and TK1 enzymatic activity were parallel in their increase and decrease. TK1 is involved in G1-S phase transition of cell cycle progression. In cell cycle analysis, we showed that radiation induced G2 arrest in K562 cells but it was not able to sustain. However, the addition of genistein to irradiated cells sustained a prolonged G2 arrest up to 120 h. In addition, the expression of cell cycle-related proteins, cyclin A and cyclin B1, provided the evidences of G1/S progression and G2-arrest, and their relationship with TK1 in cells treated with radiation and genistein. These results suggest that the activation of TK1 may be critical to modulate the radiation-induced cell death and cell cycle progression in irradiated K562 cells. PMID- 15353127 TI - The involvement of XPC protein in the cisplatin DNA damaging treatment-mediated cellular response. AB - Recognition of DNA damage is a critical step for DNA damage-mediated cellular response. XPC is an important DNA damage recognition protein involved in nucleotide excision repair (NER). We have studied the XPC protein in cisplatin DNA damaging treatment-mediated cellular response. Comparison of the microarray data from both normal and XPC-defective human fibroblasts identified 861 XPC responsive genes in the cisplatin treatment (with minimum fold change > or = 1.5). The cell cycle and cell proliferation-related genes are the most affected genes by the XPC defect in the treatment. Many other cellular function genes, especially the DNA repair and signal transduction-related genes, were also affected by the XPC defect in the treatment. To validate the microarray data, the transcription levels of some microarray-identified genes were also determined by an RT-PCR based real time PCR assay. The real time PCR results are consistent with the microarray data for most of the tested genes, indicating the reliability of the microarray data. To further validate the microarray data, the cisplatin treatment-mediated caspase-3 activation was also determined. The Western blot hybridization results indicate that the XPC defect greatly attenuates the cisplatin treatment-mediated Caspase-3 activation. We elucidated the role of p53 protein in the XPC protein DNA damage recognition-mediated signaling process. The XPC defect reduces the cisplatin treatment-mediated p53 response. These results suggest that the XPC protein plays an important role in the cisplatin treatment mediated cellular response. It may also suggest a possible mechanism of cancer cell drug resistance. PMID- 15353128 TI - Enhancement of human ACAT1 gene expression to promote the macrophage-derived foam cell formation by dexamethasone. AB - In macrophages, the accumulation of cholesteryl esters synthesized by the activated acyl-coenzyme A:cholesterol acyltransferase-1 (ACAT1) results in the foam cell formation, a hallmark of early atherosclerotic lesions. In this study, with the treatment of a glucocorticoid hormone dexamethasone (Dex), lipid staining results clearly showed the large accumulation of lipid droplets containing cholesteryl esters in THP-1-derived macrophages exposed to lower concentration of the oxidized low-density lipoprotein (ox-LDL). More notably, when treated together with specific anti-ACAT inhibitors, the abundant cholesteryl ester accumulation was markedly diminished in THP-1-derived macrophages, confirming that ACAT is the key enzyme responsible for intracellular cholesteryl ester synthesis. RT-PCR and Western blot results indicated that Dex caused up-regulation of human ACAT1 expression at both the mRNA and protein levels in THP-1 and THP-1-derived macrophages. The luciferase activity assay demonstrated that Dex could enhance the activity of human ACAT1 gene P1 promoter, a major factor leading to the ACAT1 activation, in a cell-specific manner. Further experimental evidences showed that a glucocorticoid response element (GRE) located within human ACAT1 gene P1 promoter to response to the elevation of human ACAT1 gene expression by Dex could be functionally bound with glucocorticoid receptor (GR) proteins. These data supported the hypothesis that the clinical treatment with Dex, which increased the incidence of atherosclerosis, may in part due to enhancing the ACAT1 expression to promote the accumulation of cholesteryl esters during the macrophage-derived foam cell formation, an early stage of atherosclerosis. PMID- 15353129 TI - Characterization of function of three domains in dishevelled-1: DEP domain is responsible for membrane translocation of dishevelled-1. AB - Wnt signaling plays an important role in embryogenesis and tumorgenesis. Although the mechanism about how Wnts transduce their signaling from receptor frizzled (Fz) to cytosol has not been understood, dishevelled (Dvl) protein was considered as the intersection of Wnt signal traffic. In this study, we characterized the function of three domains (DIX, PDZ and DEP) of Dvl-1 in canonical Wnt signal transduction and Dvl-1 membrane translocation. It was found both DIX and DEP domain were sufficient to block Wnt-3a-induced LEF-1 transcriptional activity and free cytosol beta-catenin accumulation; whereas PDZ domain and a functional mutant form of DEP domain (DEP-KM) had no effect on canonical Wnt signaling. In addition, when cotransfected with Fz-7, DEP domain, but not DIX, PDZ or DEP-KM, translocated and co-localized with Fz-7 to the plasma membrane, which was similar to Dvl-1. Furthermore, it was DEP domain that could block Fz-7-induced membrane translocation of Dvl-1 via a possible competitive mechanism. These results strongly suggest that DEP domain is responsible for the membrane translocation of Dvl-1 protein upon Wnt signal stimulation. PMID- 15353130 TI - Expression of DNA-dependent protein kinase in human granulocytes. AB - Human polymorphonuclear leukocytes (PMN) have been reported to completely lack of DNA-dependent protein kinase (DNA-PK) which is composed of Ku protein and the catalytic subunit DNA-PKcs, needed for nonhomologous end-joining (NHEJ) of DNA double-strand breaks. Promyelocytic HL-60 cells express a variant form of Ku resulting in enhanced radiation sensitivity. This raises the question if low efficiency of NHEJ, instrumental for the cellular repair of oxidative damage, is a normal characteristic of myeloid differentiation. Here we confirmed the complete lack of DNA-PK in PMN protein extracts, and the expression of the truncated Ku86 variant form in HL-60. However, this degradation of DNA-PK was shown to be due to a DNA-PK-degrading protease in PMN and HL-60. In addition, by using a protease-resistant whole cell assay, both Ku86 and DNA-PKcs could be demonstrated in PMN, suggesting the previously reported absence in PMN of DNA-PK to be an artefact. The levels of Ku86 and DNA-PKcs were much reduced in PMN, as compared with that of the lymphocytes, whereas HL-60 displayed a markedly elevated DNA-PK concentration. In conclusion, our findings provide evidence of reduced, not depleted expression of DNA-PK during the mature stages of myeloid differentiation. PMID- 15353131 TI - Apoptosis in granulosa cells during follicular atresia: relationship with steroids and insulin-like growth factors. AB - It is well known that during mammalian ovarian follicular development, the majority of follicles undergo atresia at various stages of their development. However, the mechanisms controlling this selection process remain unknown. In this study, we investigated apoptosis in granulosa cells during goat follicular atresia by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The changes in the levels of steroids, insulin-like growth factors (IGFs) and IGF receptors were studied by radioimmunoassay (RIA) and semi quantitative reverse transcription-PCR. We found that the percentage of apoptotic granulosa cells in the atretic (A) follicles was significantly higher than that in the slightly atretic (SA) and healthy (H) follicles. The level of estradiol and the ratio of estradiol to progesterone in H follicles were significantly higher than those in A follicles. On the other hand, the level of progesterone was not significantly different among these follicle types. We also found that the level of IGF-I in H follicles was higher than in SA and A follicles, whereas the amount of IGF-II did not vary significantly. The expression of IGF receptor also decreased in A follicles as compared to that in H and SA follicles. These results suggested that estradiol and IGF-I might be involved in controlling apoptosis in granulosa cells during follicular atresia. PMID- 15353132 TI - Pakistan and the millennium development goals for health: a case of too little, too late ? PMID- 15353133 TI - Hyperhomocysteinemia as a risk factor for ischemic heart disease. AB - OBJECTIVE: To determine association of hyperhomocysteinemia with myocardial infarction and conventional risk factors for ischemic heart disease. DESIGN: A non-interventional comparative case control study. PLACE AND DURATION OF STUDY: The study was conducted at the department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Rawalpindi and Armed Forces Institute of Cardiology / National Institute of Heart Diseases, Rawalpindi from January 2001 to June 2001. PATIENTS AND METHODS: A total of 100 hospitalized patients having myocardial infarction (MI) were randomly selected comprising 85 males and 15 females. The average age of the patients was 53 +/-4.5 years. A similar number of age and gender-matched healthy controls were also selected. The demographic details, history and clinical examination of both patients and controls were recorded and their blood was collected in fasting state for estimation of serum total cholesterol, plasma glucose and serum total homocysteine. RESULTS: Serum total homocysteine level in controls was significantly lower (10.8 +/- 4.1 micromol/L) as compared to patients (18.0 +/- 5.9 micromol/L) (p < 0.0001). Smoking showed statistically significant association with hyperhomocysteinemic patients (p = 0.04). CONCLUSION: Ischemic heart disease was associated with moderate hyperhomocysteinemia in our patients and it was also associated with smoking. PMID- 15353134 TI - Allogeneic peripheral blood stem cell transplantation in patients with haematological malignancies. AB - OBJECTIVE: To report the initial data on allogeneic peripheral blood stem cell transplantation for haematological malignancies in Pakistan. DESIGN: A single centre descriptive study. PLACE AND DURATION OF STUDY: Bismillah Taqee Institute of Health Sciences and Blood Diseases Centre from September 1999 to June 2004. PATIENTS AND METHODS: Patients with haematological malignancies were included who had received allogeneic PBSC transplantation of Filgrastim (rhG-CSF) mobilized peripheral blood stem cells from HLA-identical siblings (except one 5/6 antigen sibling) with Busulphan and Cyclophosphamide standard conditioning therapy in all patients. No patient received antibiotics for gut decontamination. Empirical antibiotics included Ceftriaxone and Amikacin for febrile neutropenia, oral Itraconazole for antifungal prophylaxis while oral acyclovir was used for antiviral prophylaxis. All donors and recipients were CMV IgG positive Cyclosporin A / Methotrexate were given for graft versus host disease (GvHD) prophylaxis. Stem cells were harvested using Haemonetics MCS+ cell separator. All patients received G-CSF starting from day +4 until their neutrophil count rose to normal. RESULTS: There were 21 patients with age range of 8-38 years and male to female ratio of 2:1. Engraftment was achieved in all patients; median time to absolute neutrophil count of > 0.5 x 10(9)/l was 10 days (range 8 - 12 days) and platelet count of > 20 x 10(9)/l was 14 days (12-17 days). Acute graft versus host disease ( aGvHD) was seen in 7 patients; one patient had grade IV skin and hepatic GvHD; another patient had grade III gut GvHD, grade II GvHD was seen in 3 patients while grade I skin aGvHD was seen in 2 patients. Median hospital stay was 34 days. Treatment related mortality was seen in 3 patients (18%). Chronic GvHD was seen in 5 patients. Four more patients died during the follow-up period. Malaria was seen in 2 while tuberculosis developed in one case. Relapse was seen in 2 patients. The estimated probability of survival at one hundred day, at one year and five years was 82, 47 and 40 percent respectively. CONCLUSION: Haematopoietic stem cell transplant programme can be developed in a developed country setting. Post transplant complications are similar to what have been reported in the developed countries. In endemic areas malaria could prove to be fatal if not recognised and treated early. PMID- 15353135 TI - Frequency of Helicobacter pylori antibodies in porto-systemic encephalopathy. AB - OBJECTIVE: To study the frequency of Helicobacter pylori antibodies in patients presenting with porto-systemic encephalopathy due to liver disease. STUDY DESIGN: An observational, cross-sectional study. PLACE AND DURATION OF STUDY: Different medical units at Civil Hospital, Karachi between May 2001 and April 2002. PATIENTS AND METHODS: During the study period, seventy-six patients of porto systemic encephalopathy due to liver diseases were selected. These subjects were evaluated for hepatic encephalopathy grade, modified Child-Pugh classification and were managed according to the standard practices. These patients were evaluated for Helicobacter (H. pylori ) antibody status by ELISA(Abbott Laboratories) method. RESULTS: Out of 76 patients studied and tested for H. pylori antibodies, 48(63.2%) were males and 28(36.8%) were females with age ranging between 17 and 85 years. Out of 76 patients who presented with porto systemic encephalopathy, 59(77.6%) had a positive H. pylori antibody test. Thirty five of these were males and 24 were females. A significant number of patients who presented with higher grade of encephalopathy were H. pylori antibody positive (p < 0.001). CONCLUSION: In this study, frequency of H. pylori antibodies was significantly high in patients of porto-systemic encephalopathy. PMID- 15353136 TI - General seroprevalence of hepatitis C and hepatitis B virus infections in population. AB - OBJECTIVE: To determine the prevalence of hepatitis C virus (HCV) and hepatitis B virus (HBV) infection by detection of anti-HCV and hepatitis B surface antigen (HbsAg) in general population of Pakistan. DESIGN: Observational. PLACE AND DURATION OF STUDY: Community clinic of Shifa International Hospital, Islamabad, from January 1998 to June 2004. MATERIALS AND METHODS: Sera of healthy adult individuals who presented for medical evaluation as a pre-employment criteria in the Gulf region were examined for presence of hepatitis B surface antigen and anti-HCV antibody. Alanine aminotransferase levels were also determined. RESULTS: A total of 47,538 individuals were examined. Out of these, 2528 (5.31%) were positive for anti-HCV and 1221 (2.56%) individuals had positive HBsAg. Hepatitis B surface antigen and anti-HCV both were found in 92 (0.19%) individuals. Mean age of subjects, positive for HCV antibody was 44 years and 40.5 years for HBV. Ninety-four percent individuals were males and 6% were females. Alanine aminotransferase (ALT) was normal in 56 % of subjects with positive HCV and 84% of individuals with HBV. CONCLUSION: This study which evaluated predominantly a healthy male population, showed a high seroprevalence of anti-HCV and average seroprevalence of hepatitis B virus infection. A large majority of these patients was young and had normal ALT. PMID- 15353137 TI - Benign tumours of the vulva. AB - OBJECTIVE: To present clinicopathological analysis of benign tumours of the vulva. DESIGN: A case series. PLACE AND DURATION OF STUDY: The study was carried out in two phases at JPMC, Karachi from October 1995 to April 1997 and at Usman Memorial Hospital from 1st February 2003 to 30th July 2003. PATIENTS AND METHODS: Thirty cases of benign tumours of vulva were studied during 2 years research period. Detailed history along with complete local and general physical examination followed by all necessary pre-operative investigations were carried out. Excision surgery was the treatment of choice in majority of cases while marsupialization was done for Bartholin's cyst. Histopathology of tumours specimen was also collected. RESULTS: A total of 30 cases were studied. Twenty two were cystic and 8 were solid tumours. Aggressive angiomyxoma was 10% of solid tumours and Bartholin's cyst was 46.6% of cystic tumours. Most of the patients were multipara and between 21-30 years of age. The main site of tumour was labium majus. Excision surgery for all cases and marsupialization for Bartholin's cyst was treatment of choice. CONCLUSION: Aggressive angiomyxoma is the commonest solid benign vulval tumour. It should be considered in the differential diagnosis of vulval mass in women of reproductive age. PMID- 15353138 TI - Management of gestational trophoblastic tumours: a five-year clinical experience. AB - OBJECTIVE: An analysis of a 5-year clinical experience in the management of gestational trophoblastic tumours in a tertiary care hospital. DESIGN: A prospective cohort follow-up study. PLACE AND DURATION OF STUDY: The study was conducted at Hayatabad Medical Complex, Peshawar from 1998 to 2003. PATIENTS AND METHODS: A total of 30 cases were managed and a detailed analysis of these patients was done. Of these 13 followed Hydatidiform Mole, 10 after abortion and 7 after a term pregnancy. RESULTS: Out of 30 cases of gestational trophoblastic tumour, 63.3% were between 21 and 38 years of age. Ninety percent of the patients presented with vaginal bleeding, while life-threatening hemorrhage occurred in 23.3%of the cases.43.3% of the patients had hydatidiform mole as an antecedent pregnancy and 36.7% of the patients presented within four months of the antecedent pregnancy. Blood groups O and B were most frequently encountered i.e. in 40% and 33.3% of the cases. Metastatic disease was present in 46.6% of the cases, of which 8 were high risk and one was of medium risk group. Major sites of metastasis were lungs (33.3%) and vagina (30%). Serum BHCG of 40,000 miu / ml and above was present in 53.3% of the cases (P=0.016) and number of metastasis >8 were found in 16.7% cases (P=0.001). Prior chemotherapy was given in only 2 patients and both of them died due to resistance. Chemotherapy was given to 100% of patients; survival was 100% in low-risk group and 50% in high-risk group (P=0.004). Overall mortality was 20% i.e. 6 patients died of the disease. Major side effects of chemotherapy were stomatitis (66.6%), alopecia (56.6%), low hemoglobin (60%), weight loss and recurrent infection. CONCLUSION: Late diagnosis, previously failed chemotherapy and high WHO prognostic scores are major risk factors affecting outcome in these patients. Hence every female in reproductive age group with unexplained bleeding per vaginum should be investigated with serum BHCG (Beta human chorionic gonadotrophin). PMID- 15353139 TI - Perinatal mortality rate in relation to gender. AB - OBJECTIVE: To calculate the gross and gender adjusted perinatal mortality rate (PMR) and determine the association between gender and susceptibility towards perinatal death. DESIGN: A cross-sectional hospital-based study conducted at Gynae Unit-I of Bolan Medical Complex Hospital, Quetta from 1st January to 31st December, 2002. PATIENTS AND METHODS: The study included the total births and perinatal deaths over one year period. The cause of death was ascertained through specifically-designed questionnaires and external autopsy. PMR was defined as the number of fetal deaths from 28 weeks of gestations to early neonatal deaths within 7 days after birth. Birth weight of >1000 grams or 35 cms crown-heel length was considered in lieu of unknown gestational age. Multiple pregnancies and stillbirths occurring at home were excluded. Extended Wiggles Worth classification was used to study the etiology of perinatal death separately. RESULTS: The PMR was calculated to be 113 per 1000 births. Stillbirth rate was 103 per 1000 total births; out of these, 56.5% were intrapartum and 43.3% antepartum. Male stillbirths were 89.24% in intrapartum and 62.5% in antepartum (df=1,X2=16, p < 0.001). Out of 16 early neonatal deaths, 11 were male infants. Aberdeen classification revealed obstetric causes of PMR as antepartum haemorrhage (34 cases;76% males), malpresentations (28 cases; 57% males), and congenital anomalies (26 cases; 80% males), after adjusting for maternal age and parity. CONCLUSION: This study shows fetal gender as statistically significant risk factor and it will pave the way for future community-based studies to confirm such an association, after adjusting for other co-variables. PMID- 15353140 TI - Central venous catheter-related bloodstream infections in cancer patients. AB - OBJECTIVE: To determine the frequency of central venous catheter-related bloodstream infections (CR-BSI) in cancer patients and the antimicrobial susceptibility pattern of the isolates. DESIGN: A cross-sectional study. PLACE AND DURATION OF STUDY: The study was conducted between April 2002 and March 2003 at the Armed Forces Institute of Pathology, Rawalpindi in collaboration with Oncology Units of Armed Forces Bone Marrow Transplant Centre and Combined Military Hospital, Rawalpindi. SUBJECTS AND METHODS: Cancer patients requiring short or long-term central venous catheterization at the time of admission or thereafter were included. Catheter tips on removal were cultured quantitatively; specimens of blood and pus were cultured qualitatively. Isolates were identified and antimicrobial susceptibility testing was performed by standard techniques. RESULTS: Eighty-nine patients were included in the study. The frequency of CR-BSI was 17%. Out of the 19 organisms isolated, 10 (53%) were Gram-positive cocci, 8 (42%) were Gram-negative rods and 1 (5%) was a fungus. Coagulase negative staphylococci (27%) were the predominant pathogens. Among the staphylococci, 46% of the isolates were methicillin-resistant. All Gram-positive isolates were susceptible to glycopeptides. Gram-negative rods were resistant to most of the commonly used antimicrobial groups. CONCLUSION: Central venous catheter is an important source of bloodstream infections in cancer patients. Most of the infections are caused by Gram-positive cocci. Rigorous infection control measures and continuous surveillance are required to curb the frequency of these infections. PMID- 15353141 TI - External fixation in close comminuted femoral shaft fractures in adults. AB - OBJECTIVE: To evaluate the role of external fixation in the management of highly comminuted femoral shaft fractures in adults. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Agency HQ Hospital, Landikotal and Saidu Group of Hospitals, Swat, from April 1999 to March 2002. PATIENTS AND METHODS: Fourteen adults with close Winquist Type III and IV comminuted fractures of the shaft of the femur were treated with external fixation. The fixator was applied until the fracture became stable and was followed by the application of a femoral cast brace till consolidation. All patients were followed for a minimum period of one year after the index procedure. RESULTS: Thirteen patients (93%) achieved fracture consolidation in an average time of 5.2 months (range 4-8). Eight patients had pin-tract infections and 3 developed shortening and 11 lost an average of 34 (0-75) degrees of range-of-motion in their knees. On the basis of this loss, there were 4 excellent, 6 good, 3 fair and 1 poor result. CONCLUSION: External fixation is a useful technique for the stabilization of close comminuted femoral shaft fractures where modern technology is not available. Pin-tract infection and knee stiffness are problems inherent with the use of this method but can be easily minimized with good pin hygiene and knee exercises. PMID- 15353142 TI - Use of norethisterone oenanthate and predisposition to thrombosis. AB - OBJECTIVE: To observe the effects of progestin-only hormonal contraceptives in our population. DESIGN: Cohort study. PLACE AND DURATION OF STUDY: Department of Pharmacology, Ziauddin Medical University, Karachi and Department of Pathology, Faculty of Health Sciences, The Aga Khan University, Karachi from October 2002 to September 2003. PATIENTS AND METHODS: The study comprised of 50 women of reproductive age group, divided into two equal and matched groups of 25 each, namely, a group of users of progestin-only injectable hormonal contraceptive Norethisterone oenanthate [NET EN] 200 mg used as a 2-monthly injection and a control group not using any hormonal contraception. Complete blood counts, prothrombin time (PT) and international normalized ratio (INR) as well as activated partial thromboplastin time (APTT) were monitored. RESULTS: APTT and prothrombin time were not significantly different in both the groups. Menstrual irregularities were found out by more frequent in users of injectable hormonal contraceptives than in non-users (p= 0.001). Hemoglobin (p= 0.029) and hematocrit were slightly better preserved in the same group. CONCLUSION: Use of Norethisterone oenanthate does not predispose to thrombogenesis in current dose for contraception. PMID- 15353143 TI - Multiple familial trichoepithelioma: a rare cutaneous tumour. AB - Multiple familial trichoepithelioma (MFT) is a rare autosomal dominant skin disease that present as many small tumours predominantly on the face. We report a case of multiple familial trichoepithelioma occurring in three members of a family. They were diagnosed simultaneously. Only one was treated with medium depth chemical peeling with partial response. PMID- 15353144 TI - A case of pulmonary sarcoidosis, not tuberculosis. AB - The case of sarcoidosis of the lungs is described in a 50 years old female who presented with symptoms similar to pulmonary tuberculosis. As she showed no improvement on anti-tubercular therapy, sarcoidosis was suspected and confirmed by endobronchial biopsy and she responded well to treatment. PMID- 15353145 TI - Postobstructive pulmonary edema associated with biting a laryngeal mask airway. AB - Laryngeal mask airway (LMA) has established itself as a reliable mode of maintaining a patent airway; however, it is associated with a few complications. This case report describes the presentation, diagnosis, management and treatment of an unusual complication associated with the use of LMA, that is postobstructive pulmonary edema (POPE), which is a potentially life-threatening complication of acute airway obstruction. It develops rapidly but responds quickly to the therapy. PMID- 15353146 TI - Hand-assisted laparoscopic surgery for colorectal malignancies. AB - OBJECTIVE: To report our initial experience with hand-assisted laparoscopic surgery (HALS) for colorectal malignancies using a specially-designed laparoscopic hand cannula. PATIENTS AND METHODS: Nine caucasians patients with colorectal malignancies underwent HALS which included 02 right hemicolectomies, 01 transverse colectomy, 03 sigmoid colectomies, 01 anterior resection and 02 low anterior resections. RESULTS: There were 4 males and 5 females. The mean length of incision for placement of the cannula was 7 cms (range 7-8 cms). The mean operating time was 180 minutes. Postoperatively on an average patients were ambulatory by day 2 (range 1-4) and taking oral fluids by day 3 (range 1-4). There were no conversions to laparotomy. Furthermore there was no operative mortality and no complication directly related to the use of the device. CONCLUSION: HALS appears to be a useful adjuvant for laparoscopic colectomy due to advantages provided by tactile sensation. A curative resection for malignancy can be performed without compromising oncological principles. PMID- 15353147 TI - Screening for hepatocellular carcinoma. AB - Liver cancer remains one of the most common causes of cancer death globally, and its cure rate has not improved for the past 20 years. Hepatocellular carcinoma develops in patients with cirrhosis of any etiology. Patients with cirrhosis are thus usually included in screening program aiming to achieve early detection and effective treatment for HCC. Only those patients diagnosed with HCC who would benefit with the available treatment, should undergo the screening procedure. This is based on ultrasonography and alpha-fetoprotein every 3-6 months based on the prevalence and economic status of the community being tested. PMID- 15353148 TI - Fetal MRI : reviewing the history, indications, technique, safety and drawbacks. AB - The ultrasound is undoubtedly the first-line diagnostic examination to identify prenatal congenital abnormalities. Fetal-MRI (FMRI) is a non-invasive, fast, highly informative examination that can shed immense information on anatomical abnormalities and affect decision-making. If, in spite of a significant pathology, the parents insist on continuing with the pregnancy, help of other interdisciplinary specialities can be sought for postnatal care. The article briefly describes different aspects of FMRI. PMID- 15353149 TI - Regional block nerve locator and insulated needle modification. PMID- 15353150 TI - Sensitivity and spectrum of bacterial isolates in infectious otitis externa. PMID- 15353151 TI - Commercial sponsorship of CME: there are alternatives. PMID- 15353152 TI - Many unnecessary Pap smears are performed after hysterectomy. PMID- 15353153 TI - Nonsurgical treatment is effective for carpal tunnel syndrome. PMID- 15353154 TI - D-dimer useful for excluding deep vein thrombosis and pulmonary embolism. PMID- 15353155 TI - Favorable response to proton pump inhibitor doesn't necessarily diagnose GERD. PMID- 15353156 TI - Liposuction does not improve cardiovascular risk factors. PMID- 15353157 TI - Changes in recommended treatments for mild and moderate asthma. PMID- 15353158 TI - How does HIPAA affect public health reporting? PMID- 15353159 TI - Resting injured limbs delays recovery: a systematic review. AB - OBJECTIVES: Rest is commonly used as primary treatment, rather than just palliation, for injured limbs. We searched the literature for evidence of benefit or harm from immobilization or mobilization of acute limb injury in adults. DATA SOURCES: We systematically searched for and retrieved randomized controlled trials (RCTs) of mobilization or rest for treatment of acute limb injuries, in Medline (1966-2002), EMBASE, Web of Science, and the Cochrane library, in all languages. REVIEW METHODS: We examined patient-centered outcomes (pain, swelling, and cost), functional outcomes (range of motion, days lost from work) and complications of treatment. RESULTS: Forty-nine trials of immobilization for soft tissue injuries and fractures of both upper and lower limbs were identified (3366 patients). All studies reported either no difference between rest and early mobilization protocols, or found in favor of early mobilization. Reported benefits of mobilization included earlier return to work; decreased pain, swelling, and stiffness; and a greater preserved range of joint motion. Early mobilization caused no increased complications, deformity or residual symptoms. CONCLUSIONS: We should not assume any benefit for immobilization after acute upper or lower limb injuries in adults. Rest appears to be overused as a treatment. More trials are needed to identify optimal programs for early mobilization. PMID- 15353160 TI - Excoriations and ulcers on the arms and legs. PMID- 15353161 TI - Interpretation of survival curves. PMID- 15353162 TI - R-E-S-P-E-C-T: patient reports of disrespect in the health care setting and its impact on care. AB - OBJECTIVE: The health care encounter is a setting in which racial/ethnic disparities can arise. Patients who experience disrespect in this encounter may be less likely to use health care services that improve outcomes. The objective of this study was to examine factors in the health care encounter and to model how negative perceptions of the encounter influence health care utilization. DESIGN, SUBJECTS, AND SETTING: Data were derived from the Commonwealth Fund 2001 Health Care Quality Survey, a nationwide random-digit-dial survey of 6722 adults, conducted between April 30 and November 5, 2001. Bivariate and multivariate analyses were performed on weighted data. MAIN OUTCOME MEASURES: Measures of negative perceptions of the patient-provider relationship included feelings of being treated with disrespect or being looked down upon, a belief that unfair treatment was received due to race or language spoken, and a belief that better treatment would have been received had the respondent had been of a different race. Measures of utilization included receipt of a physical exam within the prior year, receipt of optimal cancer screening and recommended elements of chronic disease care, delay of needed care, and not following the doctor's advice. MAIN RESULTS: Minorities were significantly more likely to report being treated with disrespect or being looked down upon in the patient-provider relationship. Specifically, 14.1% of blacks (P=.06), 19.4% of Hispanics (P<.001), and 20.2% if Asians (P<.001) perceived being treated with disrespect or being looked down upon, compared with only 9.4% of whites. Persons who thought that they would have received better treatment if they were of a different race were significantly less likely to receive optimal chronic disease screening and more likely to not follow the doctor's advice or put off care (P<.01). CONCLUSIONS: Perceptions of disrespect or of receiving unfair treatment within the patient provider relationship are prevalent, particularly among racial/ethnic minorities. Such negative perceptions influence health care utilization and may contribute to existing health disparities. PMID- 15353163 TI - Clinical inquiries. Do routine eye exams reduce occurrence of blindness from type 2 diabetes? PMID- 15353164 TI - Clinical inquiries. What is the most effective diagnostic evaluation of streptococcal pharyngitis? PMID- 15353165 TI - Clinical inquiries. Is the long-term use of proton pump inhibitors safe? PMID- 15353166 TI - Clinical inquiries. First- or second-generation antihistamines: which are more effective at controlling pruritus? PMID- 15353167 TI - Clinical inquiries. What is the most effective way for relieving constipation in children aged >1 year? PMID- 15353168 TI - Taste-aversion-prone (TAP) rats and taste-aversion-resistant (TAR) rats differ in ethanol self-administration, but not in ethanol clearance or general consumption. AB - Taste-aversion (TA)-prone (TAP) rats and TA-resistant (TAR) rats have been developed by means of bidirectional selective breeding on the basis of their behavioral responses to a TA conditioning paradigm. The TA conditioning involved the pairing of an emetic-class agent (cyclophosphamide) with a novel saccharin solution as the conditioned stimulus. Despite the absence of ethanol in the selective breeding process, these rat lines differ widely in ethanol self administration. In the current study, blood alcohol concentrations (BACs) were determined after 9 days of limited (2 h per day) access to a simultaneous, two bottle choice of a 10% ethanol in water solution [volume/volume (vol./vol.)] or plain water. The BACs correlated highly with ethanol intake among TAR rats, but an insufficient number of TAP rats yielded measurable BACs to make the same comparison within this rat line. The same rats were subsequently exposed to 24-h access of a two-bottle choice (10% ethanol or plain water) for 8 days. Ethanol consumption during the 24-h access period correlated highly with that seen during limited access. Subsequent TA conditioning with these rats yielded line-typical differences in saccharin preferences. In a separate group of rats, ethanol clearance was determined by measuring BACs at 1, 4, and 7 h after injection of a 2.5-g/kg dose of ethanol. Ethanol clearance was not different between the two lines. Furthermore, the lines did not differ with respect to food and water consumption. Therefore, the TAP rat-TAR rat differences in ethanol consumption cannot be attributed to line differences in ethanol metabolism or in general consummatory behavior. The findings support our contention that the line differences in ethanol consumption are mediated by differences in TA-related mechanisms. The findings are discussed with respect to genetically based differences in the subjective experience of ethanol. PMID- 15353169 TI - Ethanol increases tumor necrosis factor-alpha receptor-1 (TNF-R1) levels in hepatic, intestinal, and cardiac cells. AB - Chronic ethanol consumption leads to cell injury in virtually every tissue. Tumor necrosis factor-alpha (TNF-alpha) constitutes a major factor in the development of alcohol-induced liver injury. In alcohol-dependent subjects, elevated levels of plasma TNF-alpha are strongly predictive of mortality. Binding of TNF-alpha to TNF-alpha receptor-1 (TNF-R1) activates death domain pathways, leading to necrosis and apoptosis in most tissues, and it also increases the expression of intercellular adhesion molecules (i.e., ICAM-1), which promote inflammation. We determined whether ethanol exposure leads to increases in cellular TNF-R1. We incubated HepG2 human hepatoma cells and H4-II-E-C3 rat hepatoma cells with 25, 50, and 100 mM ethanol for various intervals of time up to 48 h. Human colonic adenocarcinoma cells (Caco-2 cells) and neonatal rat primary cardiomyocytes were also incubated with different concentrations of ethanol. Levels of TNF-R1 were measured either by a sandwich enzyme-linked immunosorbent assay (ELISA) method or by determining the extracellular transmembrane domain of TNF-R1 by an intact-cell ELISA method. Ethanol exposure for 48 h increased TNF-R1 levels in human hepatoma cells in a dose-dependent manner. Levels increased significantly by 164% at 50 mM and by 240% at 100 mM ethanol. Effects were time dependent and did not reach a plateau at 48 h. Similar increases in TNF-R1 were also observed in rat hepatoma cells (90% at 50 mM and 230% at 100 mM ethanol). Under similar conditions, Caco-2 cells showed a significant 80% increase in TNF-R1 levels at 200 mM ethanol, a concentration found in intestine. Neonatal rat primary cardiomyocytes showed TNF R1 increases of 36% at 50 mM and 44% at 100 mM ethanol. These results indicate that exposure of different cell types to pharmacologic concentrations of ethanol increases TNF-R1 levels and may augment TNF-alpha-mediated cell injury in different tissues. PMID- 15353170 TI - Gene expression in the liver of rats fed alcohol by means of intragastric infusion. AB - It has become increasingly evident that one of the most fruitful approaches to understanding cellular processes and their relation to disease consists of large scale gene profiling of cells, tissues, and organs. This also constitutes a first step in exploring the molecular biologic basis of various diseases. In the current study, we used cDNA microarray technology to assess possible changes in the expression of a large number of genes in the liver of rats fed alcohol (ethanol) chronically (4 weeks) by means of intragastric infusion. This animal model resembles closely the alcoholic liver disease in human beings. Of a total of 8,740 probe sets arrayed on the microchip, 2,069 were expressed by the liver. After a correction for false discovery rate at 10%, 72 genes were found to be significantly up-regulated (40) or down-regulated (32). Forty-two genes were suppressed, and four genes were induced, by alcohol. These genes are involved in a wide spectrum of cellular functions. Also, the genes that underwent significant changes were categorized into two groups: genes that have been implicated in alcoholic liver disease and genes that have not been tested for possible changes in expression. Large-scale gene profiling of the liver reveals changes in the expression of a number of genes that have never been implicated in alcohol induced injury. Further investigation of such genes may cast light on mechanisms underlying alcohol-induced liver injury and help in the design of new therapeutic approaches. PMID- 15353171 TI - Maternal separation has no effect on voluntary ethanol intake in female Wistar rats. AB - Adverse experiences, early in life or during adulthood, can increase the vulnerability for development of drug dependence. Investigators have shown that short and prolonged periods of maternal separation during the postnatal period can affect voluntary ethanol intake in male rats. Recent study findings have indicated sex-dependent effects of maternal separation, and, in the current study, the effects of maternal separation on acquisition of voluntary ethanol intake in female Wistar rats were investigated. Rat pups were subjected to daily maternal separation for 15 min (MS15) or 360 min (MS360) during the first 3 weeks of life or reared under normal animal facility rearing (AFR) conditions. During 25 days, starting at 10 weeks of age, the effects of maternal separation on acquisition of voluntary ethanol intake were investigated. In contrast with previous study results for male rats, neither MS15 nor MS360 affected acquisition of voluntary ethanol intake in female rats. A stressful situation in adulthood, restraint stress, resulted in a significant increase in ethanol intake during the restraint period compared with baseline levels in the animals reared under normal AFR conditions, an effect that persisted throughout the postrestraint period. In rats subjected to MS15 or MS360, a significant increase in ethanol intake was shown during the postrestraint period compared with baseline levels. The current study findings therefore give further evidence for sex differences in the consequences of maternal separation. Compared with previous findings in male rats, acquisition of ethanol intake was not affected, and restraint-induced effects were less pronounced but more prolonged, in female rats. PMID- 15353172 TI - Effects of a low dose of ethanol in an animal model of premenstrual anxiety. AB - Low (1 mM), but not 10 mM, concentrations of ethanol selectively potentiate current gated by alpha(4)beta(2)delta subunit combinations of the gamma aminobutyric acid type A (GABA(A)) receptor, a subtype increased in hippocampus after withdrawal from progesterone in a rodent model of premenstrual anxiety. In the current study, we tested the hypothesis that the anxiolytic effect of ethanol would exhibit a similar dose-response effect by using the acoustic startle response (ASR) and elevated plus-maze as behavioral models. To this end, adult, female rats were tested (1) 24 h after removal of a progesterone-filled capsule implanted subcutaneously for 21 days (progesterone withdrawal) or (2) on the day of diestrus, a low hormone state. Low doses of ethanol (0.2-0.4 mg/kg) produced a significant 60%-70% decrease in the ASR only in animals undergoing progesterone withdrawal. However, higher doses of ethanol (0.8-1.2 g/kg) were ineffective in these animals, resulting in an "inverted U" ethanol dose effect similar to that observed at recombinant alpha(4)beta(2)delta subunit combinations of the GABA(A) receptor. Consistent with these findings, significant 70% attenuation of the ASR was also achieved after progesterone withdrawal with 3 mg/kg of 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP), a GABA(A) receptor partial agonist with greater potency at alpha(4)betadelta receptors than at other known isoforms. In contrast, this partial agonist was not anxiolytic in control animals. These results support the suggestion that very low doses of ethanol are anxiolytic in a model of premenstrual anxiety, whereas higher, potentially sedative, doses are without effect. The results may be relevant for altered ethanol sensitivity during premenstrual syndrome, when increased ethanol consumption has been reported. PMID- 15353173 TI - Effects of chronic naltrexone treatment in rats on place preference and locomotor activation after acute administration of cocaethylene or ethanol plus cocaine. AB - When cocaine and ethanol are taken together a cocaine metabolite called cocaethylene is produced. Investigators have determined that cocaine, ethanol, and cocaethylene all produce a conditioned place preference when administered intraperitoneally. On the basis of the moderate success of naltrexone at attenuating the rewarding effects of ethanol and cocaine administered independently, we examined the ability of chronic naltrexone treatment (administered by means of subcutaneous implant across 6 days) to reduce the preference and motor-stimulating effects resulting from intraperitoneal administration of cocaethylene (Experiment 1) and the co-administration of ethanol with cocaine (Experiment 2) in outbred rats. Results demonstrated naltrexone modestly reduced conditioned place preference for cocaethylene but had no effect on the locomotor stimulation resulting from cocaethylene administration. Naltrexone failed to decrease the preference for the chamber paired with co-administration of ethanol and cocaine and did not change the degree of locomotor activation produced by these drugs. These results support the suggestion that naltrexone as a pharmacotherapy to treat co-abuse of ethanol and cocaine in human beings may have limited benefits. PMID- 15353174 TI - Exercise reverses ethanol inhibition of neural stem cell proliferation. AB - Neural stem cells in the adult brain may contribute to learning and neural plasticity, to stress-induced neuropathologic changes, to mood and affective disorders, and to other complex brain functions. Learning, an enriched environment, and exercise (as modeled by running) all increase neural stem cell proliferation in the hippocampus and improve performance on learning tests, whereas acute ethanol consumption decreases neural stem cell proliferation in the hippocampus. To explore the interaction of exercise and ethanol consumption (two behaviors important for mental health), C57BL/6 mice were given access to ethanol, a running wheel, or both, and neural stem cell proliferation was investigated in the hippocampal dentate gyrus, corpus callosum, and forebrain subventricular zone of these mice. Mice given access to ethanol consumed large amounts of ethanol, and mice given access to a running wheel ran long distances, with the combined groups' behavior being comparable to that of the ethanol access alone group and of the running wheel access alone group. Neural stem cell proliferation was assessed by treating mice with bromodeoxyuridine (12 days, 300 mg/kg/day, i.p.) and evaluating consequent immunoreactivity. Exercise increased bromodeoxyuridine immunoreactivity in dentate gyrus, but not in other brain regions studied. Self-administration of ethanol decreased bromodeoxyuridine immunoreactivity by approximately 60% (P <.01) in dentate gyrus; 70% (P <.001), in forebrain; and 80% (P <.001), in corpus callosum. Findings for exercise and ethanol consumption combined were remarkably similar to those for exercise alone. The opposing effects of ethanol consumption and exercise on neurogenesis could contribute to the CNS pathology and health benefits, respectively, of these two behaviors. PMID- 15353175 TI - Genomic heterogeneity and instability of the AZF locus on the human Y chromosome. AB - The spermatogenesis locus azoospermia factor (AZF) in Yq11 has been mapped to three microdeletion intervals designated as AZFa, AZFb, and AZFc. They are caused by intrachromosomal recombination events between large homologous repetitive sequence blocks, and AZFc microdeletions are now recognised as the most frequent known genetic lesion causing male infertility. However, in the same Y-region, large genomic heterogeneities are also observed in fertile men, and only complete AZFa and AZFb deletions are associated with a specific testicular pathology. Partial AZF deletions are associated with variable pathologies and partial AZFc deletions may even have no impact on male fertility. This suggests a genetic redundancy of the multi-copy genes in AZFb and AZFc and a causative relationship between the occurrence of first microdeletions then macrodeletions in the repetitive structure of Yq11 where large palindromes are probably promoting multiple gene conversions and AZF rearrangements. PMID- 15353176 TI - Nuclear receptor LXRalpha is involved in cAMP-mediated human renin gene expression. AB - The cAMP-signaling pathway plays a crucial role in the regulation of the renin gene, but the mechanism involved remains poorly understood. We have focused our studies of renin gene regulation on the unique cAMP responsive element (huREN/CNRE, -135 to -107) in the human renin promoter. We have cloned a protein that binds to this unique CNRE and demonstrated that this protein is liver X receptor-alpha (LXRalpha), a transcriptional factor of the nuclear receptor family. Transient expression of LXRalpha in human renin-producing Calu-6 cells increased cAMP inducibility of human renin promoter. Similarly, LXRalpha-stably transfected Calu-6 cells exhibited increased cAMP inducibility of renin promoter as well as the endogenous renin gene. Site-directed mutation of huREN/CNRE, which disrupted LXRalpha binding, decreased cAMP-induced transcriptional activity of human renin promoter. Furthermore, we demonstrated that the binding of LXRalpha derived from human juxtaglomerular cells, the main production site of renin in the kidney, to the huREN/CNRE in vivo. These results suggest that LXRalpha plays an important role in the cAMP-mediated regulation of human renin gene transcription by binding to CNRE. PMID- 15353177 TI - Ciliary neurotrophic factor influences endocrine adipocyte function: inhibition of leptin via PI 3-kinase. AB - Ciliary neurotrophic factor (CNTF), originally known for its involvement in the modulation of neuronal growth, has been discovered to exert anorexigenic effects and is currently being investigated in clinical studies for the treatment of obesity and insulin resistance. This neuropeptide acts on the central nervous system. However, we have recently demonstrated direct peripheral effects on adipocyte signalling and thermogenesis. Given the emerging endocrine role of adipose tissue in the regulation of energy homeostasis and insulin resistance, we investigated potential effects of CNTF on leptin expression and secretion. Our study demonstrates a direct inhibition of leptin expression and secretion by acute and chronic CNTF treatment. Furthermore, we demonstrate a differentiation- and Janus kinase 2 (JAK2)-independent, but phosphatidylinositol 3-kinase dependent signalling pathway mediating this negative effect. These results provide novel evidence for a role of CNTF in the selective modulation of adipocyte endocrine function which may have important implications for the regulation of energy homeostasis. PMID- 15353178 TI - Autocrine regulation of steroidogenic function of Leydig cells by transforming growth factor-alpha. AB - We have determined the effects of LH on the expression of transforming growth factor-alpha (TGFalpha) and epidermal growth factor receptor (EGFR) system in rat Leydig cells and investigated its role in steroidogenesis. LH and TGFalpha/epidermal growth factor (EGF) significantly increased the levels of TGFalpha mRNA and protein, and the levels of EGFR protein in immature rat Leydig cells (ILC). Treatment with TGFalpha or EGF for 24h resulted in significant increase in androgen production in ILC. The increase in androgen production in response to TGFalpha was associated with increased mRNA levels of SR-BI, steroidogenic acute regulatory (StAR) and P450scc but not of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and P450c17. TGFalpha also caused a marked increase in the levels StAR protein in ILC. EGFR inhibitor (AG1478) blocked the effects of TGFalpha while MEK-inhibitor (PD98059) potentiated TGFalpha or LH effects on steroidogenesis. A PKA inhibitor (H89) blocked both TGFalpha and LH effects on steroidogenesis. We conclude that TGFalpha plays an autocrine role in LH dependent development and function of Leydig cells. PMID- 15353179 TI - Spontaneously immortalized epithelial cells from mouse caput epididymidis. AB - We report here on the characterization of tissue-culture cell lines derived from primary cultures of the mouse caput epididymidis epithelium. The cell lines were spontaneously immortalized without the use of transforming oncogenes. In defined conditions, our epididymal cells adopted various morphological features that resembles that of the in vivo epididymis epithelium such as a polarized organization and the presence of junctional structures at their apical/lateral membranes as revealed by electron microscopy analyses. Flow cytometry analysis revealed that we were dealing with homogenous cell populations that had reached a near-tetraploid state. RT-PCR assays were used in order to show that several genes that can be considered as markers of in vivo caput epididymidis epithelium activity were expressed in our cell lines confirming that these cells were indeed in a differentiated state close to their endogenous state. PMID- 15353180 TI - Receptor-selective determinants in catfish gonadotropin seat-belt loops. AB - Mammalian gonadotropins are highly selective. Charge differences between the Cys(10-11) sequence of FSHbeta and LHbeta/CGbeta seat-belt loops determine the ability of these hormones to interact with the LH-R. Selective FSH-R binding is mainly dependent on the presence of an FSHbeta-specific sequence between Cys(11 12) of the seat-belt loop. Intriguingly, African catfish LHbeta (cfLHbeta) lacks a positively charged Cys(10-11) region and stimulates both catfish LH-R and FSH-R with comparable potencies. Our studies on the promiscuous behaviour of cfLH using chimeric gonadotropins revealed that the Cys(10-11) region of cfLHbeta contains cfLH-R-selective determinants, whereas the Cys(11-12) region of cfLHbeta confers FSH-R-stimulating activity to cfLH. Hence, the location of receptor-selective determinants appeared to be fairly well conserved throughout evolution, despite the low sequence identity between mammalian and catfish seat-belt loops. Moreover, various structure-function differences between gonadotropins are discussed in the context of the different (female) reproductive strategies between mammalian and non-mammalian species that required the divergence to a more specific LH-R-stimulating activity of one of the gonadotropins in mammals. PMID- 15353181 TI - Proteasome-dependent degradation of ERalpha but not ERbeta in cultured mouse aorta smooth muscle cells. AB - Here we investigate ERalpha and ERbeta expression and regulation in vascular smooth muscle cells from mouse aorta. Immunocytochemistry showed nuclear staining for both ERalpha and ERbeta. Double stainings revealed co-expression of ERalpha and ERbeta in vascular smooth muscle cells. ERalpha (66 kDa) and ERbeta (54 kDa) expression determined by Western blotting was unchanged within 7 h after inhibition of protein synthesis with cycloheximide in the absence of 17beta estradiol (E(2)), showing that both proteins are stable without ligand-binding. Treatment with 10 nM E(2) for 7 h in the presence of cycloheximide increased ERalpha, suggesting that E(2) causes a conformational change in the ERalpha protein. The ERbeta was not affected by E(2). Treatment with the proteasome inhibitor epoxomicin (100 nM) for 3 days caused a prominent upregulation of ERalpha both in the absence and in the presence of E(2), while ERbeta was unaffected, suggesting that ERalpha but not ERbeta is degraded by ubiquitin proteasome system in vascular smooth muscle cells. In summary, we disclose a short-term regulation of ERalpha protein by estrogen and that ERalpha but not ERbeta is degraded via the ubiquitin-proteasome pathway in vascular smooth muscle cells. PMID- 15353182 TI - Evaluation of the role of FSH in regulation of Leydig cell function during different stages of its differentiation. AB - The role of follicle stimulating hormone (FSH) in Leydig cell function was evaluated by a passive neutralization approach at different stages of Leydig cell development. Neutralization of endogenous FSH in neonatal rats (10-day-old) resulted in reduction of testes weight, however the testicular testosterone levels and in vitro testosterone production by purified Leydig cells were elevated. Administration of FSH antiserum to immature (25-28-day-old) and adult (90-day-old) rats did not have any effect on testes weight, serum testosterone and testicular testosterone. Interestingly, there was a significant reduction in testosterone production by isolated Leydig cells under hCG stimulated and 22-R hydroxycholesterol (22-R-OH CHOL) saturated conditions. In support of this observation administration of recombinant FSH to immature and adult rats resulted in significant increase in testosterone production by Leydig cells following incubations in presence of hCG and saturating concentrations of 22-R-OH CHOL, although there was no change in serum and testicular testosterone levels. The role of FSH in immature rats was also confirmed employing FSH receptor antiserum which was raised against the unique domains of FSH receptor. RT-PCR analysis revealed a significant reduction in the mRNA levels of StAR and IGF-1 following blockade of FSH action by FSH receptor antiserum. The results of our studies suggest a stage specific function for FSH in regulation of Leydig cell development by modulating the LH responsiveness and steroidogenesis. PMID- 15353183 TI - A pint a day raises a man's pay; but smoking blows that gain away. AB - This paper studies the wage effects of the use of alcohol and tobacco. The analysis based on a recent survey in The Netherlands shows that for males the use of tobacco has a negative wage effect of about 10% while the use of alcohol has a positive wage effect of about the same size. The wages of females are not affected by smoking and drinking. PMID- 15353184 TI - On the possibility of a bridge between CBA and CEA: comments on a paper by Dolan and Edlin. AB - In a recent work by Dolan and Edlin, it is concluded that no link can be established between cost-benefit analysis (CBA) and cost-effectiveness analysis (CEA). However, the conclusion seems to depend rather heavily on what is understood by a link between CBA and CEA as well as on the exact meaning of the latter two terms. We argue that there is at least one approach to CBA and CEA in which the two are very intimately linked. On the other hand, the limitations in the access to preference information has consequences for the kind of questions that can be meaningfully addressed in both CBA and CEA. PMID- 15353185 TI - Anti-social welfare functions: a reply to Hansen et al. AB - We could reasonably expect society to give at least the same weight to the marginal utility of the poor as to the rich, and to the marginal utility of the ill as compared to the healthy. Whilst Hansen et al. [Journal of Health Economics (2004)], may be said to link CEA and CBA within a welfarist framework, the assumptions they require are inconsistent with these types of ethical preferences. Thus, the degree to which they employ a reasonable social welfare function is doubtful. This paper argues that any link between CEA and CBA will occur not within a welfarist framework but instead within a non-welfarist one in which it is unlikely that CBA results could be easily transformed into cost effectiveness ratios. PMID- 15353186 TI - Obesity and risk knowledge. AB - Obesity is an epidemic health problem in many developed countries, and it is an emerging public health concern in developing, transitional, and newly developed countries. The purpose of this research is to investigate the relationship between individuals' knowledge concerning the health risks of obesity and their tendency to be obese (as measured by the "body mass index"). Instead of assuming that obesity is a pure physiological problem as in previous studies, we allow an individual's cost/benefit evaluation to play a role. Based on survey data from Taiwan, we investigate the relationship with the quantile regression technique. The results suggest that such a relationship does exist and it is different for males and females. PMID- 15353187 TI - To err on humans is not benign. Incentives for adoption of medical error reporting systems. AB - Concerns about frequent and harmful medical errors have led policy makers to advocate the creation of a system for medical error reporting. Health providers, fearing that reported information about errors would be used against them under the current medical malpractice system, have been reluctant to participate in such reporting systems. We propose a re-design of the malpractice system -- one in which penalties are a function of the health provider's reporting efforts -- to overcome this incentive problem. We also consider some alternatives to this mechanism that address two important ways in which reporting effort may not be observable: hospitals may have interests distinct from individual physicians and may not be able to observe their reporting efforts, and a regulatory agency or a court may not be able to adequately observe reporting efforts by a provider. PMID- 15353188 TI - The effect of HMOs on fee-for-service health care expenditures: evidence from Medicare revisited. AB - This paper implements a new method for calculating the extent of selection in the aged Medicare HMO market. Selection is measured as the difference in average costs between new Medicare HMO enrollees and Medicare fee-for-service stayers with data from 1990 to 1994. Results suggest Medicare HMO enrollees were 1030 US dollars cheaper in their first year of enrollment. The effect is found entirely in Part A (hospital) expenditures, confirming selection is based on inpatient rather than outpatient or preventive care. These results are consistent with previous work. PMID- 15353189 TI - Socio-economic status, health and lifestyle. AB - The role of lifestyle in mediating the relationship between socio-economic characteristics and health has been discussed extensively in the epidemiological and economic literatures. Previous analyses have not considered a formal framework incorporating unobservable heterogeneity. In this paper, we develop a simple economic model in which health is determined (partially) by lifestyle, which depends on preferences, budget and time constraints and unobservable characteristics. We estimate a recursive empirical specification consisting of a health production function and reduced forms for the lifestyle equations using Maximum Simulated Likelihood (MSL) for a multivariate probit (MVP) model with discrete indicators of lifestyle choices and self-assessed health (SAH) on British panel data from the 1984 and 1991 Health and Lifestyle Survey (HALS). We find that sleeping well, exercising, and not smoking in 1984 have dramatic positive effects on the probability of reporting excellent or good SAH in 1991, and that these effects are much larger having accounted for endogeneity. The failure of epidemiological analyses to account for unobserved heterogeneity can explain their low estimates of the relevance of lifestyle in the socio-economic status-health relationship. Indicators for prudent alcohol consumption and eating breakfast in 1984 are not found to be statistically significant determinants of SAH in 1991. PMID- 15353190 TI - Innovation and risk selection in deregulated social health insurance. AB - One important motive for deregulating social health insurance is to encourage product innovation. For the first time, the cost savings achieved by non-US managed care plans that are attributable to product innovation are estimated, using a novel approach. Panel data from a major Swiss health insurer permits to infer health status, which can be used to predict health care expenditure. The econometric evidence suggests that the managed care plans benefit from risk selection effects. In the case of the health maintenance organization (HMO) plan, however, the pure innovation effect may account for as much as two-thirds of the cost advantage. PMID- 15353191 TI - The transition from good to poor health: an econometric study of the older population. AB - This is a study of the influence of socioeconomic factors on the state of health of older Canadians. Three years of panel data from the Survey of Labour and Income Dynamics are used to model the transition probabilities between good and poor health. Care is taken to avoid the problem of endogeneity of income in modelling its effects, and to adjust reported income to free it from its strong association with age at the time of the survey. Of particular note are the significant effects found for income, in spite of universal public health care coverage. Significant effects are found also for age, education, and other variables. PMID- 15353192 TI - Parallel imports and the pricing of pharmaceutical products: evidence from the European Union. AB - We consider policy issues regarding parallel imports (PIs) of brand-name pharmaceuticals in the European Union, where such trade is permitted. We develop a simple model in which an original manufacturer competes in its home market with PI firms. The model suggests that for small trade costs the original manufacturer will accommodate the import decisions of parallel traders and that the price in the home market falls as the volume of parallel imports rises. Using data from Sweden we find that the prices of drugs subject to competition from parallel imports fell relative to other drugs over the period 1994-1999. Econometric analysis finds that parallel imports significantly reduced manufacturing prices, by 12-19%. There is evidence that this effect increases with multiple PI entrants. PMID- 15353193 TI - The effect of the State Children's Health Insurance Program on health insurance coverage. AB - This paper presents the first national estimates of the effects of the SCHIP expansions on insurance coverage. Using CPS data on insurance coverage during the years 1996-2000, we estimate instrumental variables regressions of insurance coverage. Our regression results imply that 9% of children meeting income eligibility standards for SCHIP gained public insurance. While low, our estimates indicate that states were more successful in enrolling children in SCHIP than they were with prior Medicaid expansions that were focused on children just above the poverty line. Crowd-out of private health insurance was estimated to be nearly 50%, which is in line with estimates for the Medicaid expansions of the early 1990s. In addition, state anti-crowd-out provisions in the form of waiting periods were found to significantly affect both take-up and crowd-out. PMID- 15353194 TI - Corrosion behavior and surface characterization of titanium in solution containing fluoride and albumin. AB - The objective of this study was to demonstrate the role of albumin on the corrosion behavior of titanium in a solution containing 2.0 g/l fluoride and either 0.1 or 1.0 g/l albumin. The corrosion behavior and surface characterization of passive films on titanium immersed in such a solution were examined. In addition, the change in pH and the concentration of dissolved titanium in the solution were examined. The results showed that the corrosion of titanium in a solution containing fluoride was distinct, and that adding albumin to the solution containing fluoride suppressed corrosion. Fluorine was detected on the titanium surface immersed in the solution containing fluoride, and dissolution of the titanium was confirmed. The titanium immersed in a solution containing both fluoride and albumin had an albumin film regardless of the albumin concentration. In addition, the amount of dissolved titanium from the titanium immersed in the solution was less than when the solution contained no albumin. It was suggested that the formation of adsorbed albumin films on or in the passive film acted to not only protect the titanium from attack by the fluoride but also suppressed dissolution of the titanium-fluoride compounds. PMID- 15353195 TI - Potential of biomimetic surfaces to promote in vitro osteoblast-like cell differentiation. AB - Bioactive glasses, osteoproductive materials, have received considerable attention as bone graft substitutes in the treatment of bony defects. More recent strategies for achieving a predictable periodontal regeneration include the use of enamel matrix proteins, due to their role in the formation of bone tissue. The aim of our study is to examine the effects of these materials on the proliferation and differentiation of the mouse preosteoblastic cell line MC3T3 E1. Cells were cultured up to 28 days in contact with three types of granules: Bioglass 45S5 granules (BG), 45S5 granules coated with enamel matrix proteins (Emdogain) (BG/EMD), and a less reactive glass used as a control (60S). Phase contrast microscopic observations have shown that all substrates supported the growth of osteoblastic cells. Zones of differentiation were observed at an earlier stage in cultures of BG and BG/EMD. TEM observations revealed ultrastructural features very close to what is observed in vivo during intramembranous ossification with a direct bone apposition on the bioactive glasses. Total protein production was higher in the cultures with BG and BG/EMD. Northern Blot analysis revealed a stimulation of the transcription factor Cbfa1/Runx2 at day 13 in cultures of BG when compared to the two other cultures. Bone sialoprotein (early marker of differentiation) and osteocalcin (marker of late-stage differentiation) expression was increased in cultures with BG and BG/EMD when compared to 60S. Taken together, our findings indicate that Bioglass alone or combined with Emdogain, have the ability to support the growth of osteoblast-like cells in vitro and to promote osteoblast differentiation by stimulating the expression of major phenotypic markers. In addition, we noticed that the bioactive granules coated with Emdogain revealed significantly higher protein production than the bioactive granules alone at day 20. PMID- 15353196 TI - Quantitative assessment of the response of osteoblast- and macrophage-like cells to particles of Ni-free Fe-base alloys. AB - In the present study, the effect of mechanically alloyed particles of new FeAlCr alloys developed for potential applications as surgical implants has been tested on osteoblast- and macrophage-like cells and compared to particles of the Ti6Al4V alloy, for which there is a good clinical experience. After microstructural characterisation of the particles, cells were cultured with particles for 24-48 h using three different concentrations of particles, and the response of cells was quantified by assessment of viability, proliferation, and morphology. Mineralisation by osteoblasts was verified after 21 days. The amount of aluminium and chromium ions in the culture medium of macrophages was measured by graphite furnace atomic absorption and phagocytosis of particles assessed by light microscopy. Viability and proliferation of osteoblast- and macrophage-like cells were substantially unaffected by the presence of particles of the new alloys, which were phagocytosed according to their size. Aluminium and chromium ions were released in the culture medium, but no direct correlation with the cell behaviour was found. In vitro mineralisation was achieved by osteoblasts in due time. The new alloys are well tolerated in in vitro systems, and, due to their chemical and mechanical characteristics, they are under development for surgical implants. PMID- 15353197 TI - Whole blood coagulation on protein adsorption-resistant PEG and peptide functionalised PEG-coated titanium surfaces. AB - The aim of this study was to investigate whole blood coagulation on low blood plasma protein adsorbing surfaces. For this purpose, the polycationic graft copolymer poly(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG), PLL-g-PEG grafted with a cell adhesive peptide containing the amino acid sequence -Arg-Gly-Asp- (RGD), and PLL-g-PEG with a control peptide -Arg-Asp-Gly- (RDG) were adsorbed onto titanium (oxide), forming stable monomolecular adlayers through electrostatic attraction. Free oscillation rheometry and complementary techniques were used to measure the coagulation time (CT) and other interactions of the surfaces with native whole blood, recalcified platelet-rich plasma (PRP), and recalcified citrated platelet-free plasma (PFP). The results show that the uncoated titanium surfaces (reference) activated platelets and quickly triggered the coagulation cascade via the intrinsic pathway, whereas the PLL-g-PEG surfaces displayed a prolonged CT, approximately 2-3 times longer compared to uncoated titanium. We hypothesise that blood coagulates outside the vascular system independent of low protein adsorption to or activation by surfaces, due to the absence of an active down-regulation of procoagulative processes by the vascular endothelium. PMID- 15353198 TI - Intracellular protein phosphorylation in adherent U937 monocytes mediated by various culture conditions and fibronectin-derived surface ligands. AB - Macrophages play a central role in the normal healing process after tissue injury and the host response to foreign objects such as biomaterials. The process leading to macrophage adhesion and activation on protein-adsorbed substrates is complex and unresolved. While the use of primary cells offers clinical relevancy, macrophage cell lines offer unique advantages such as availability and relatively homogeneous phenotype as models to probe the molecular mechanism of cell-surface interaction. Our goal was to better characterize the effect of the culture condition and surface-associated ligands on the extent of U937 adhesion. Tyrosine phosphorylation of intracellular proteins was surveyed as a basis to seek a greater understanding of the molecular mechanism involved in mediating U937 adhesion on various ligand-adsorbed surfaces. U937 viability and adhesion on tissue culture polystyrene (TCPS) increased with (i) increasing serum level, (ii) decreasing tyrosine phosphorylation inhibitor AG18 concentration, or (iii) increasing culture time. The adsorption of various adhesion proteins such as fibronectin and peptide ligands (i.e., RGD, PHSRN) on TCPS did not significantly increase the adherent density of U937 when compared with albumin and PBS ligand controls. However, ligand identity and the presence of phorbol myristate acetate dramatically affected the extent (i.e., increase or decrease) and the identity (i.e., molecular weight) of phosphotyrosine proteins in adherent U937 in a time dependent manner. The extent and identity of phosphotyrosine proteins did not exhibit a clear AG18 dose dependency, rather the level of tyrosine phosphorylation for a distinct group of proteins was either increased or decreased for a given AG18 concentration. PMID- 15353199 TI - Patterned networks of mouse hippocampal neurons on peptide-coated gold surfaces. AB - Patterned networks of hippocampal neurons were generated on peptide-coated gold substrates prepared by microscope projection photolithography and microcontact printing. A 19 amino acid peptide fragment of laminin A (PA22-2) that includes the IKVAV cell adhesion domain was used to direct patterns of cell adhesion in primary culture. Microscale grid patterns of peptide were deposited on gold coated glass cover slips by soft lithography using "stamps" fashioned from polydimethylsiloxane. Strong coordination bonding between gold atoms on the surface and the sulfur atoms of the N-terminal cysteine residues supported stable adhesion of the peptide, which was confirmed by immunofluorescence using anti IKVAV antiserum. Dispersed hippocampal cells isolated from neonatal mouse pups were grown on peptide-patterned gold substrates for 7 days. Neurons preferentially adhered to peptide-coated regions of the gold surface and restricted their processes to the peptide patterns. Whole cell recordings of neurons grown in patterned arrays revealed an average membrane potential of -50 mV, as well as the presence of voltage-gated ion conductances. Peptide-modified gold surfaces serve as convenient and effective substrates for growing ordered neural networks that are compatible with existing multi-electrode array recording technology. PMID- 15353200 TI - Adhesion contact dynamics of primary hepatocytes on poly(ethylene terephthalate) surface. AB - The design of bioartificial liver assist device requires an effective attachment of primary hepatocytes on polymeric biomaterials. A better understanding of this cell-surface interaction would aid the optimal choice of biomaterials. In this study, the adhesion contact dynamics of primary hepatocytes on poly(ethylene terephthalate) (PET) surface with grafted poly(acrylic acid) (PAA) and coated collagen is probed with confocal reflectance interference contrast microscopy (C RICM) in conjunction with phase contrast microscopy. An increase of acrylic acid density from 0 to 12 nmole/cm2 raises both the root-mean-square surface roughness and amount of adsorbed collagen of PET surface. C-RICM demonstrates that hepatocytes form tight adhesion contacts upon seeding on both plain PET and PAA grafted PET (both with collagen coating) despite the insignificant two dimensional cell spreading. At two hours after cell seeding, the normalized contact area and adhesion energy of hepatocytes on 12 nmole/cm2 PAA-grafted-PET (with collagen coating) is 27% and 114% higher, respectively, than that on collagen coated plain PET. Interestingly, the growth kinetics of adhesion patch for hepatocyte on PAA-grafted PET with collagen coating is best fitted by R proportional to t0.5 and is significantly different from that on collagen coated plain PET, which is best fitted by R proportional to t0.25. Overall, this study demonstrates the modulation of biophysical response of adherent hepatocytes through the control of the biomaterial surface properties. PMID- 15353201 TI - Evaluation of three-dimensional scaffolds made of blends of hydroxyapatite and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) for bone reconstruction. AB - Hydroxyapatite (HAP) was blended into poly(3-hydroxybutyrate) (PHB) and poly(3 hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) to make films and scaffolds. After HAP blending, mechanical properties of PHB including compressive elastic modulus and maximum stress showed improvement and osteoblast responses including cell growth and alkaline phosphatase activity were also strengthened. On the other hand, scaffolds made of PHBHHx blended with HAP had an adverse effect. No remarkable change on degradation of PHB or PHBHHx blended with HAP, respectively, was observed in simulated body fluid. Scanning electron microscopy examination revealed that osteoblast responses to HAP incorporation may be related to surface morphology and to the exposed HAP particles on polymer surface. All these results indicated that the blending of HAP particles into PHBHHx scaffolds fabricated by salt leaching was unable to either strengthen its mechanical properties or enhance osteoblast responses. Although HAP is bioactive and osteoconductive, its blending with PHBHHx did not generate a better performance on bone reconstruction. PMID- 15353202 TI - Deformation, yielding, fracture and fatigue behavior of conventional and highly cross-linked ultra high molecular weight polyethylene. AB - Medical grade ultra high molecular weight polyethylene (UHMWPE) has been used as the bearing surface of total joint replacements for over four decades. These polymeric devices are susceptible to accumulated cyclic damage in vivo. Wear debris formation that ultimately leads to a need for revision surgery is linked to the plasticity, fatigue and fracture mechanisms of UHMWPE. This paper examines the deformation, yielding, fracture and fatigue behavior of conventional and highly cross-linked medical grade UHMWPE. Such properties play an important role in determining the long-term success of orthopedic devices. The mechanical properties discussed include the deformation behavior of UHMWPE, the yielding associated with quasi-static tension and compression, fracture toughness, cyclic loading, and fatigue resistance. PMID- 15353203 TI - Nitric oxide-releasing sol-gels as antibacterial coatings for orthopedic implants. AB - To assess the benefits of nitric oxide (NO)-releasing sol-gels as potential antibacterial coatings for orthopedic devices, medical-grade stainless steel is coated with a sol-gel film of 40% N-aminohexyl-N-aminopropyltrimethoxysilane and 60% isobutyltrimethoxysilane. Upon converting the diamine groups in these films to diazeniumdiolate NO donors, the NO release from the sol-gel-coated stainless steel is evaluated at both ambient and physiological temperature. Sol-gel films incubated at 25 degrees C have a lower NO flux over the first 24 h compared to those at 37 degrees C, but release more than five times longer. The bacterial adhesion resistance of NO-releasing coatings is evaluated in vitro by exposing bare steel, sol-gel, and NO-releasing sol-gel-coated steel to cell suspensions of Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis at 25 degrees C and 37 degrees C. Cell adhesion to bare and sol-gel-coated steel is similar, while NO-releasing surfaces have significantly less bacterial adhesion for all species and temperatures investigated. PMID- 15353204 TI - Surface characteristics of holmium-loaded poly(L-lactic acid) microspheres. AB - Radioactive holmium-166-loaded poly(L-lactic acid) microspheres (Ho-PLLA-MS) are promising systems for the treatment of liver malignancies. The surface characteristics of Ho-PLLA-MS before and after both neutron and gamma irradiation were investigated in order to get insight into their suspending behaviour and to identify suitable surfactants for clinical application of these systems. X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) were used for surface characterization. The residual amounts of poly(vinyl alcohol) (PVA) of the microspheres, which was used as an emulsifier during the solvent evaporation process, were determined using a colorimetric iodine-borate method and the wettability of microspheres and PLLA films with and without holmium (Ho) loading was tested using suspending experiments and contact angle measurements. XPS showed that the surface of Ho-PLLA-MS mainly consisted of PLLA, less than 10% of the surface was covered with PVA after several washing and sieving steps. A colorimetric assay showed that the microspheres contained 0.2-0.3% (w/w) PVA. Combined with XPS data, this assay demonstrates that the PVA is likely dissolved in the core of the microspheres. XPS analysis also showed that after neutron irradiation, some holmium appeared on the surface. Moreover, Ho-loaded PLLA films had a much higher contact angle (85 degrees) than non-loaded films (70 degrees). Therefore, the Ho on the surface of neutron-irradiated Ho-PLLA-MS is probably the reason for their poor suspending behaviour in saline. No surface changes were seen with XPS after gamma irradiation. Based on their surface characteristics, a pharmaceutically acceptable solvent (1% Pluronic F68 or F127 in 10% ethanol) was formulated with which a homogeneous suspension of radioactive Ho-PLLA-MS could be easily obtained, making these systems feasible for further clinical evaluation. PMID- 15353205 TI - Physico-chemical studies of the gelation of chitosan in a hydroalcoholic medium. AB - The formation of chitosan physical hydrogels without any external cross-linking agent was studied. This gelation took place in an acetic acid-water-propanediol solution. Static light scattering was used to detect the gel point and then, to study the gelation for different initial conditions. Thus, we investigated the influence of the degree of acetylation, the gelation temperature and the nature of the initial solvent. The variation of the solvent composition during gelation was determined from a simple weighting, and the ionisation state of the polymer at the gel point, by pH titrations. This work showed that it was possible to form a chitosan physical-hydrogel, whatever the degree of acetylation provided typical conditions were observed. The mechanism of gelation simply consisted in the modification of the hydrophilic/hydrophobic balance allowing the formation of both hydrophobic interactions and hydrogen bonding. Several parameters had an important role on this mechanism: 1--the apparent charge density of chitosan, modified by the degree of neutralisation, 2--the dielectric constant of the solvent, related to the composition of the medium, 3--the degree of acetylation, 4--temperature, playing a role on the interactions responsible for the physical cross-linking and the molecular mobility, and, 5--the molecular mobility depending on possible changes of conformation, steric hindrance and viscosity of the medium. PMID- 15353206 TI - Modeling of dispersed-drug release from two-dimensional matrix tablets. AB - A mathematical model was developed and analytical solutions were obtained for dispersed-drug release from two-dimensional matrix tablets in a perfect sink. This model can be used to describe kinetics of solute release from matrices with isotropic or anisotropic properties. Moving boundaries of dispersed-drug in both radial and axial directions and release kinetics were predicted by the model. Various factors influencing release kinetics were analyzed including the ratio of initial solute loading (C0) to solute solubility (Cs), the anisotropy of the matrix and the aspect ratio of tablet radius to the half-thickness. The model is also applicable to 1-D planar or 1-D cylindrical geometries when R/H is larger than 100 or smaller than 0.01. PMID- 15353207 TI - Impaired peroxisomal function in the central nervous system with inflammatory disease of experimental autoimmune encephalomyelitis animals and protection by lovastatin treatment. AB - Peroxisomes are ubiquitous subcellular organelles and abnormality in their biogenesis and specific gene defects leads to fatal demyelinating disorders. We report that neuroinflammatory disease in brain of experimental autoimmune encephalomyelitis (EAE) rats decreased the peroxisomal functions. Degradation of very long chain fatty acids decreased by 47% and resulted in its accumulation (C26:0, 40%). Decreased activity (66% of control) of dihydroxyacetonephosphate acyltransferase (DHAP-AT), first enzyme in plasmalogens biosynthesis, resulted in decreased levels of plasmalogens (16-30%). Catalase activity, a peroxisomal enzyme, was also reduced (37%). Gene microarray analysis of EAE spinal cord showed significant decrease in transcripts encoding peroxisomal proteins including catalase (folds 3.2; p<0.001) and DHAP-AT (folds 2.6; p<0.001). These changes were confirmed by quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis, suggesting that decrease of peroxisomal functions in the central nervous system will have negative consequences for myelin integrity and repair because these lipids are major constituents of myelin. However, lovastatin (a cholesterol lowering and anti-inflammatory drug) administered during EAE induction provided protection against loss/down-regulation of peroxisomal functions. Attenuation of induction of neuroinflammatory mediators by statins in cultured brain cells [J. Clin. Invest. 100 (1997) 2671-2679], and in central nervous system of EAE animals and thus the EAE disease [J. Neurosci. Res. 66 (2001) 155-162] and the studies described here indicate that inflammatory mediators have a marked negative effect on peroxisomal functions and thus on myelin assembly and that these effects can be prevented by treatment with statins. These observations are of importance because statins are presently being tested as therapeutic agents against a number of neuroinflammatory demyelinating diseases. PMID- 15353208 TI - The metabotropic glutamate receptor 5 is necessary for late-phase long-term potentiation in the hippocampal CA1 region. AB - Selective antagonists of the metabotropic receptors 1 (mGluR1), +/-2-methyl-4 carboxyphenylglycine (LY367385), and mGluR5, 2-methyl-6-(phenylethynyl)-pyridine (MPEP), were used to investigate the role of group I metabotropic receptors in late-phase long-term potentiation (L-LTP) at Schaffer collateral/commissural fiber-CA1 synapses in rat hippocampal slices. L-LTP was induced with three trains of tetanization of 1 s duration at 100 Hz separated by 10-min intervals. Neither LY367385 nor MPEP affected basal synaptic responses at the doses used (200 and 10 microM, respectively) and only the mGluR5 inhibitor MPEP blocked L-LTP. However, in agreement with previous mouse mutant studies, we found that both LY367385 and MPEP inhibited the induction of an LTP obtained with a single train of tetanization of 1 s duration at 100 Hz. MPEP's ability to disrupt L-LTP was not due to an effect on NMDA responses since it did not affect pharmacologically isolated N-methyl-D-aspartate (NMDA) excitatory postsynaptic potentials (EPSPs). However, MPEP prevented the increased phosphorylation in dendrites of p70 S6 kinase (p70(S6K)) at Thr3889, a major regulator of translation required for the induction of protein synthesis-dependent forms of LTP. PMID- 15353209 TI - Cells of monocyte/microglial lineage are involved in both microvessel amyloidosis and fibrillar plaque formation in APPsw tg mice. AB - Ultrastructural three-dimensional reconstruction indicates that deposition of amyloid in the wall of capillaries and in perivascular plaques in APP(SW) transgenic mice (Tg2576) represents two steps of one pathological process associated with inflammation of the vascular wall and perivascular space with cells of monocyte/microglia lineage and fibrillar amyloid-beta deposition. Plaque growth is associated with an increase in the number of microglial cells from two in the smallest plaque to 113 in the largest plaque; however, the growth in the number of microglial cells does not result in amyloid deposit degradation. On the contrary, an increase in the number and volume of microglial cells correlates with the growth of amyloid star from 62 to 34,460 microm(3), and an increase of the plaque volume from 1555 to 284,497 microm(3) (r=0.9). Growth in the number of microglial cells in the absence of morphological evidence of fibrillar amyloid internalization and phagocytosis indicates that microglial cells do not remove amyloid in Tg2576 mice. The study suggests that (a) the mechanism of capillary amyloidosis and plaque formation is similar, (b) the cells of monocyte/macrophage lineage play a critical role in fibrillar amyloid deposition in both types of lesions, and (c) treatment of one of these two forms of brain amyloidosis may affect both types of pathological changes. PMID- 15353210 TI - Transient cerebral ischemia induces site-specific hyperphosphorylation of tau protein. AB - Neurofibrillary tangles (NFTs) are a pathological hallmark of many neurodegenerative diseases including Alzheimer's disease (AD), Parkinson's disease (PD), and frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP17). However, the cellular origin and the consequence of the NFT formation are poorly understood. Epidemiological evidence suggests a much higher occurrence of dementia in stroke patients. This may represent the pathogenesis of sporadic AD, which accounts for the majority of AD occurrence. Here we show that after a transient cerebral ischemia, hyperphosphorylated tau accumulates in cortical neurons in a site-specific manner. The hyperphosphorylated tau presents a conformation similar to those present in human tauopathies, and colocalizes largely with signs of apoptosis. Our current study suggests that tau hyperphosphorylation may contribute to the brain damage induced by transient cerebral ischemia, and may be involved in the pathogenesis of neurodegenerative disorders in patients after stroke. Further, these results indicate that ischemic neuronal damage and apoptosis associates with tau hyperphosphorylation, and potentially NFTs formation. Finally, our results also suggest that neuronal apoptosis may be a therapeutic target in preventing tauopathy-related neurodegenerative diseases. PMID- 15353211 TI - Increased cholecystokinin release in the rat anterior cingulate cortex during carrageenan-induced arthritis. AB - Several human and animal studies indicate that the anterior cingulate cortex (ACC) plays an important role in the affective component of pain. The neuropeptide cholecystokinin (CCK) is especially abundant in the ACC. CCK has been suggested to be involved in the mediation of anxiety and in the modulation of opioid effects in the spinal cord and medulla oblongata. However, its possible role in pain transmission or modulation in the brain is far less clear. In this study, a model of subchronic inflammatory pain in rats, carrageenan-induced monoarthritis, was used to study the effect of pain on the release of CCK-like immunoreactivity (CCK-LI) in the ACC. Pain-related behaviour quantified by weight bearing and stance scoring, as well as inflammation measured by ankle oedema, was increased for at least 24 h after carrageenan injection with a maximum at 5 h. Using microdialysis in freely moving rats, extracellular concentrations of CCK-LI was measured in the ACC during a time period when the animals showed significant pain behaviour. In animals with carrageenan-induced arthritis, both basal and potassium-evoked release of CCK-LI were significantly increased compared to controls. HPLC analysis of dialysates from the ACC during potassium stimulation showed that the main part of the immunoreactive material was sulphated CCK-8. Because CCK has been implicated in anxiety, we suggest that an altered CCK-ergic activity in the ACC may be of importance for the affective component of pain, but an involvement in the modulation of nociception is also possible. PMID- 15353212 TI - Astrocytic proliferation in the piriform cortex of amygdala-kindled subjects: a quantitative study in partial versus fully kindled brains. AB - Complex partial epilepsy is a seizure disorder in which attacks frequently arise from foci located in the temporal lobes. The amygdala-kindling model is a widely used model of complex partial epilepsy with secondary generalization. The present study was designed to quantitatively assess astrocytic changes in the rat piriform cortex in the amygdala-kindling model of epilepsy. Bromodeoxyuridine injected subjects were sacrificed 24 h after the first stage 1 or fifth stage 5 seizure. Brain sections were prepared and examined quantitatively. A significantly higher number of dividing astrocytes (identified by co-labeling with antibodies to bromodeoxyuridine and to one of the astrocytic intermediate filament proteins glial fibrillary acidic protein or vimentin) was found in both partially kindled (stage 1) and fully kindled (stage 5) brains. The partially kindled brains had a significantly higher number of double-labeled cells on the side ipsilateral to stimulation. The opposite trend was observed in the fully kindled brains. Differences between the ipsilateral and contralateral sides of the kindled brain may suggest different role(s) for astrocytes in the development and progression of the seizure-prone state. PMID- 15353213 TI - Necrosis, apoptosis and hybrid death in the cortex and thalamus after barrel cortex ischemia in rats. AB - Focal ischemia in the cerebral cortex results in acute and delayed cell death in the ischemic cortex and non-ischemic thalamus. We examined the hypothesis that neurons in ischemic and non-ischemic regions died from different mechanisms; specifically, we tested whether a mixed form of cell death containing both necrotic and apoptotic changes could be identified in individual cells. Focal barrel cortex ischemia in rats was induced by occlusion of small branches of the middle cerebral artery (MCA) corresponding to the barrel cortex, local blood flow was measured by quantitative autoradiography. Cell death was visualized by 2,3,5 triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (H&E) staining, the terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and caspase-3 staining 1 to 10 days after the ischemia. Electron microscopy was used for ultrastructural examination. Cell death occurred in the ipsilateral cortex 24 h after ischemia, followed by selective neuronal death in the ventrobasal (VB) thalamus 3 days later. TUNEL positive neurons were found in these two regions, but with striking morphological differences, designated as type I and type II TUNEL positive cells. The type I TUNEL positive cells in the ischemic cortex underwent necrotic changes. The type II TUNEL positive cells in the thalamus and the cortex penumbra region represented a hybrid death, featured by concurrent apoptotic and necrotic alterations in individual cells, including marked caspase-3 activation, nuclear condensation/fragmentation, but with swollen cytoplasm, damaged organelles and deteriorated membranes. Cell death in the thalamus and the cortex penumbra were attenuated by delayed administration of the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone (Z-VAD-FMK). Our data suggest that TUNEL staining should be evaluated with morphological changes, the hybrid death but not typical apoptosis occurs in the penumbra region and non-ischemic thalamus after cerebral ischemia. PMID- 15353214 TI - Specific changes in cerebral second messenger accumulation underline REM sleep inhibition induced by the exposure to low ambient temperature. AB - In the rat the exposure to an ambient temperature (Ta) of -10 degrees C induces an almost total REM sleep deprivation that results in a proportional rebound in the following recovery at normal laboratory Ta when the exposure lasts for 24 h, but in a rebound much lower than expected when the exposure lasts 48 h. The possibility that this may be related to plastic changes in the nervous structures involved in the control of thermoregulation and REM sleep has been investigated by measuring changes in the concentration of adenosine 3':5'-cyclic monophosphate (cAMP) and D-myo-inositol 1,4,5-trisphosphate (IP(3)) in the preoptic-anterior hypothalamic area (PO-AH), the ventromedial hypothalamic nucleus (VMH) and, as a control, the cerebral cortex (CC). Second messenger concentration was determined in animals either stimulated by being exposed to hypoxia, a depolarizing condition that induces maximal second messenger accumulation or unstimulated, at the end of a 24-h and a 48-h exposure to -10 degrees C and also between 4 h 15 min and 4 h 30 min into recovery (early recovery). At the end of both exposure conditions, cAMP concentration significantly decreased in PO-AH-VMH, but did not change in CC, whilst changes in IP(3) concentration were similar in all these regions. The low cAMP concentration in PO-AH-VMH was concomitant with a significantly low accumulation in hypoxia. The normal capacity of cAMP accumulation was only restored in the early recovery following 24 h of exposure, but not following 48 h of exposure, suggesting that this may be a biochemical equivalent of the REM sleep inhibition observed during 48 h of exposure and which is carried over to the recovery. PMID- 15353215 TI - Endogenous neurotensin attenuates dopamine-dependent locomotion and stereotypy. AB - The neuropeptide neurotensin (NT) is highly sensitive to changes in dopaminergic signaling in the striatum, and is thought to modulate dopamine-mediated behaviors. To explore the interaction of NT with the dopamine system, we utilized mice with a targeted deletion of dopamine synthesis specifically in dopaminergic neurons. Dopamine levels in dopamine-deficient (DD) mice are less than 1% of control mice, and they require daily administration of the dopamine precursor L dihydroxyphenylalanine (L-DOPA) for survival. DD mice are supersensitive to the effects of dopamine, becoming hyperactive relative to control mice in the presence of L-DOPA. We show that 24 h after L-DOPA treatment, when DD mice are in a "dopamine-depleted" state, Nt mRNA levels in the striatum of DD mice are similar to those in control mice. Administration of L-DOPA or L-DOPA plus the L amino acid decarboxylase inhibitor, carbidopa, (C/L-DOPA) induced Nt expression in the striatum of DD mice. The dopamine D1 receptor antagonist, SCH23390, blocked C/L-DOPA-induced Nt. To test the hypothesis that this striatal Nt expression modulated dopamine-mediated behavior in DD mice, we administered SR 48692, an antagonist of the high affinity NT receptor, together with L-DOPA or C/L-DOPA. L-DOPA-induced hyperlocomotion and C/L-DOPA-induced stereotypy were potentiated by peripheral administration of SR 48692. Furthermore, intrastriatal microinjections of SR 48692 augmented L-DOPA-induced hyperlocomotion. These results demonstrate a dynamic regulation of striatal Nt expression by dopamine via D1 receptors in DD mice, and point to a physiological role for endogenous striatal NT in counteracting motor behaviors induced by an overactive dopamine system. PMID- 15353216 TI - An interhemispheric asymmetry in motor cortex disinhibition during bimanual movement. AB - The release of short interval intracortical inhibition (SICI) induced by passive bimanual movement was assessed in dominant and non-dominant primary motor cortices (M1). Dual-pulse focal transcranial magnetic stimulation (TMS) was delivered over M1 while the limbs were at rest, and during the mid-flexion phase of contralateral rhythmical wrist flexion-extension. Test and conditioned responses were recorded from flexor carpi radialis (FCR) when the wrist was passively moving alone, during bimanual mirror symmetric passive synchronous movement, and during bimanual passive asynchronous movement. Tonic inhibition was released to a greater extent in the non-dominant M1 than in the dominant M1 during synchronous mirror symmetric movement. This interhemispheric asymmetry was not evident during asynchronous movement. The findings support the idea that the dominant M1 has the capacity to disinhibit homologous representations in the contralateral M1 during synchronous movement, but the non-dominant M1 does not reciprocate to the same extent. PMID- 15353217 TI - Neurotrophic effects of bone morphogenetic protein-7 in a rat model of Parkinson's disease. AB - Previous studies have demonstrated that pretreatment with bone morphogenetic protein-7 (BMP7) reduces ischemic neuronal injury in vivo. Moreover, exogenous application of BMP7 increases both the number of tyrosine hydroxylase (+) cells and dopamine (DA) uptake in rat mesencephalic cell cultures. The purpose of this study was to investigate the in vivo effects of BMP7 on 6-hydroxydopamine (6 OHDA) induced lesioning of midbrain DA neurons. Adult Fischer 344 rats were anesthetized and injected with BMP7 or vehicle into the left substantia nigra, followed by local administration of 9 microg of 6-OHDA into the left medial forebrain bundle. The lesioned animals that received BMP7 pretreatment, as compared to vehicle/6-OHDA controls, had a significant reduction in methamphetamine-induced rotation 1 month after the surgery. BMP7-pretreatment partially preserved KCl-induced dopamine release in the lesioned striatum and significantly increased TH immunoreactivity in the lesioned nigra and striatum. In summary, our data suggest that BMP7 has neuroprotective and/or neuroreparative effects against 6-OHDA lesioning of the nigrostriatal DA pathway in an animal model of Parkinson's disease (PD). PMID- 15353218 TI - Dissociation of hyperalgesia from fever following intracerebroventricular administration of interleukin-1beta in the rat. AB - Interleukin-1beta (IL-1beta) is a cytokine that contributes to the hyperalgesia, inactivity, and fever associated with illness. These three components of the illness response occur simultaneously following peripheral administration of IL 1beta. The objective of the present study was to determine whether hyperalgesia, inactivity, and fever correspond following central administration. Rats were injected with IL-1beta (0.05 pg-50 ng/10 microl) into the lateral ventricle and core body temperature and activity were assessed for 5.5 h using radio telemetry while rats remained in their home cage. Rats were removed from the cage periodically to assess nociception by measuring the latency for hindpaw withdrawal to radiant heat. The two highest doses of IL-1beta (5 and 50 ng) caused an increase in core body temperature and a decrease in activity beginning 105 min following administration. No change in nociception was evident at any time after administration of IL-1beta regardless of dose. These data indicate that the hyperalgesia associated with fever is triggered by a peripheral, not a central action of IL-1beta, presumably by activation of vagal afferents. PMID- 15353219 TI - Deficits of neuronal density in CA1 and synaptic density in the dentate gyrus, CA3 and CA1, in a mouse model of Down syndrome. AB - Ts65Dn mice are partially trisomic for the distal region of MMU16, which is homologous with the obligate segment of HSA21 triplicated in Down syndrome (DS). Ts65Dn mice are impaired in learning tasks that require an intact hippocampus. In order to investigate the neural basis of these deficits in this mouse model of Down syndrome, quantitative light and electron microscopy were used to compare the volume densities of neurons and synapses in the hippocampus of adult Ts65Dn (n=4) and diploid mice (n=4). Neuron density was significantly lower in the CA1 of Ts65Dn compared to diploid mice (p<0.01). Total synapse density was significantly lower in the dentate gyrus (DG; p<0.001), CA3 (p<0.05) and CA1 (p<0.001) of Ts65Dn compared to diploid mice. The synapse-to-neuron ratio was significantly lower in the DG (p<0.001), CA3 (p<0.01) and CA1 (p<0.001) of Ts65Dn compared to diploid mice. When the data were broken down by synapse type, asymmetric synapse density was found to be significantly lower in the DG (p<0.001), CA3 (p<0.05) and CA1 (p<0.001) of Ts65Dn compared to diploid mice, while such a difference in symmetric synapse density was only present in the DG (p<0.01). The asymmetric synapse-to-neuron ratio was significantly lower in the DG (p<0.001), CA3 (p<0.01) and CA1 (p<0.001) of Ts65Dn compared to diploid mice, but there were no such significant differences in symmetric synapse-to-neuron ratios. These results suggest that impaired synaptic connectivity in the hippocampus of Ts65Dn mice underlies, at least in part, their cognitive impairment. PMID- 15353220 TI - Immunohistochemical and morphometrical studies on myelin breakdown in the demyelination (dmy) mutant rat. AB - The demyelination (dmy) rat is a unique mutant exhibiting severe myelin breakdown in the central nervous system (CNS). In this study, we conducted immunohistochemical and morphometrical investigations in the dmy rat. From around 6 weeks of age, the affected rats developed ataxia especially in the hindlimbs. Afterwards, ataxia worsened rapidly, resulting in complete paralysis of the hindlimbs and recumbency. Histopathology at 7 to 10 weeks of age revealed myelin destruction throughout the white matter of the CNS in the dmy rats. The most severely affected lesions were distributed in the corpus callosum, capsula interna, striatum, subcortical white matter, cerebellar peduncle, and ventral and lateral parts of the spinal cord. Immunohistochemistry demonstrated prominent astrogliosis and many ED-1 positive macrophages in the myelin-destructed areas. Until the 4th week, no significant differences in myelin thickness and fiber diameter were found between dmy and control rats. However, from 5 weeks of age, myelin thickness of residual myelinated fibers in dmy rats became significantly less than that in controls. These data indicated that the dmy phenotype shows a prolonged period of myelin destruction, suggesting that dmy mutation affects the adequate maintenance of myelin. PMID- 15353221 TI - Excitability changes in resting forearm muscles during voluntary foot movements depend on hand position: a neural substrate for hand-foot isodirectional coupling. AB - When associating hand and foot voluntary oscillations, isodirectional coupling is preferred irrespective of hand position (prone or supine). To investigate the neural correlates of this coupling modality, excitability of the motor projections innervating the resting forearm was tested during cyclic voluntary flexion-extensions of the ipsilateral foot. H-reflexes, in some experiments facilitated by subliminal Transcranial Magnetic Stimulation (TMS), and Compound Muscle Action Potentials (CMAPs), evoked by supraliminal TMS, were elicited in Flexor Carpi Radialis (FCR) and Extensor Carpi Radialis (ECR) muscles at five intervals during the foot movement cycle. With the hand prone, a sinusoidal excitability modulation was observed in wrist flexors and extensors, but reversed in phase: in FCR, excitability increased during plantar-flexion and decreased during dorsiflexion, while in ECR the opposite occurred. This reciprocal organisation was confirmed by the excitability modulation of CMAPs evoked simultaneously in the two antagonists. When the hand was supinated, the H-reflex modulation reversed in phase, i.e., FCR excitability increased during foot dorsiflexion and decreased during plantar-flexion. In both muscles and hand positions tested, when the muscle-to-movement phase-lag was increased by inertial loading of the foot, H-reflex excitability modulations remained phase linked to muscular contractions, not to movement. Together, these results suggest that the subliminal excitability modulation of hand movers has a common central origin with the parallel overt activation of foot movers, is reciprocally organised, and is direction- not muscle-dependent. It may therefore represent the neural substrate for isodirectional coupling of hand (prone or supine) with the foot. PMID- 15353222 TI - Estradiol-induced changes in the activity of hippocampal neurons in network culture are suppressed by co-incubation with gabapentin. AB - The ovarian steroid hormone estradiol, in addition to its function in the maintenance and regulation of reproductive capacity, can alter neuronal excitability. Estradiol is proconvulsant, increases neuronal excitability and decreases the threshold for seizure activity. Over one-third to one-half of women with epilepsy experience catamenial seizures, which are seizures influenced by cyclical hormone changes. These hormone-sensitive seizures respond to the anti epileptic drug gabapentin, which is a structural analogue of the inhibitory amino acid neurotransmitter GABA. We studied the effects of 17-beta-estradiol alone and estradiol co-incubated with gabapentin on neuronal activity in network cultures of rat hippocampal neurons using a fluorescent calcium binding dye fluo-3 AM, FM 1-43 labeling of synaptic vesicles and electrophysiological recordings. Significant changes in the neuronal network activity were observed in the estradiol-treated neuronal cultures; the reactivity of the neurons to KCl depolarization induced intracellular calcium changes, and FM 1-43 destaining was increased as was the frequency of spontaneous miniature excitatory postsynaptic currents (mEPSC). All these excitatory effects of estradiol were nullified by co incubating the neurons with a combination of estradiol and gabapentin. This suggests that gabapentin can indeed affect the estradiol-induced changes in neuronal network hyperexcitability by influencing the neuronal calcium levels, exocytosis and synaptic activity. Our findings could provide an understanding of the cellular basis of hormone-sensitive seizure control by gabapentin. PMID- 15353223 TI - Medullary noradrenergic neurons release norepinephrine in the medial amygdala in females in response to mating stimulation sufficient for pseudopregnancy. AB - In the female rat, stimuli from the uterine cervix and vagina are carried to the brain areas involved in the mating-induced pseudopregnancy (PSP) response via the ventral noradrenergic bundle. Noradrenergic neurons projecting through this tract synapse in many forebrain areas including the amygdala, and neurons in the posterodorsal medial amygdala (MePD) are activated following mating. The goal of this experiment was to investigate whether norepinephrine (NE) is released into the MePD after mating using microdialysis and to determine the origin of this release. Ovariectomized estrogen- and progesterone-treated rats were implanted unilaterally with guide cannulae aimed at the MePD. Females were placed with males until they received 15 intromissions (15I), 5 intromissions (5I) or 15 mounts-without-intromission (MO). Dialysate samples collected every 20 min for 2 h before to 3 h after mating were analyzed for NE using HPLC with electrochemical detection. A significant increase in mean NE release in the MePD was seen at 80 min after mating onset in females receiving 15I, and no increase was seen in animals receiving 5I or MO. The time of peak NE release varied in 15I animals from 60 to 160 min after mating. Mean baseline levels of NE did not differ between groups. The retrograde tracer FluoroGold (FG), administered through the probe after cessation of dialysis sampling, was observed within identified noradrenergic cells primarily within the A1 and A2 cell groups. Infusion of anti dopamine-beta-hydroxylase-saporin (DBH-SAP) into the MePD lesioned noradrenergic neurons located in the A1 and A2 cell groups. Because high levels of NE release occurred in the MePD only after the females received a number of intromissions sufficient to induce PSP, these results suggest that NE release within the MePD may be important for the establishment of PSP. PMID- 15353224 TI - Cholinergic neurons in the basal forebrain of aged female mice. AB - Aging is associated with at least down-regulation of several cellular functions and diminished responsiveness to internal and external signals, and possibly with direct cell death. Consequently, pharmacological manipulations may be less effective in aged than in young organisms. In the present study, we investigated whether the basal forebrain cholinergic neurons and the estrogen receptor alpha (ERalpha) which they contain respond to changes in estrogen availability in aged female mice. The mice were sham-operated, ovariectomized, or ovariectomized and treated with 17beta-estradiol at the age of 18 months. Three months later, the mice were perfused and brain sections were double immunostained for choline acetyltransferase (ChAT) and ERalpha. Cell counting with a stereological method revealed that changes in the estrogen level have no effect on the total number of ChAT-immunoreactive (ir) neurons in the basal forebrain. However, the percentage of ChAT-ir neurons containing ERalpha-ir was higher in the ovariectomized mice than in the sham-operated or estrogen-treated mice. This was specific for the medial septum and vertical diagonal band of Broca. The findings indicate that even at old age the ERalphas in cholinergic cells are able to respond to changes in estrogen levels, though in a region-specific manner. This is naturally important for studies aiming to develop therapies for the elderly. PMID- 15353225 TI - Acute effects of 17beta-estradiol on the extracellular concentration of excitatory amino acids and energy metabolites during transient cerebral ischemia in male rats. AB - Elevation of extracellular levels of amino acids has been implicated in the pathogenesis of stroke. The failure of brain energy metabolism due to the lack of oxygen and glucose contributes also to cell loss. Estrogen has been shown to protect brain cells against ischemia by a still unclear mechanism. We used intracerebral microdialysis to monitor the effects of acute 17beta-estradiol treatment on the release of glutamate and aspartate and on the levels of the energy metabolites glucose and lactate. In male rats subjected to 90 min of transient middle cerebral artery occlusion followed by 24-h reperfusion, acute treatment with 17beta-estradiol (0.8 mg/kg, i.v.) at the time of occlusion reduced the ischemic infarct by about 50%. In these treated rats, the ischemia induced increases of extracellular levels of glutamate and aspartate were significantly and rapidly reduced. The reduction of glucose level during occlusion was not affected by 17beta-estradiol treatment; however, the increase of extracellular lactate was reduced during occlusion and reperfusion, probably due to the reduced glutamate-driven astrocytic glycolysis. These data suggest that acute treatment with 17beta-estradiol at the onset of occlusion significantly reduces the ischemia-induced excitotoxicity in the cortex, a mechanism that may participate in the neuroprotective effect on cellular survival. PMID- 15353226 TI - Identification of the protein disulfide isomerase family member PDIp in experimental Parkinson's disease and Lewy body pathology. AB - Parkinson's disease (PD) is a slowly progressing neurodegenerative disorder with no clear etiology. Pathological hallmarks of the disease include the loss of dopaminergic neurons from the substantia nigra (SN) and the presence of Lewy bodies (LBs) (alpha-synuclein and ubiquitin-positive, eosinophilic, cytoplasmic inclusions) in many of the surviving neurons. Experimental modeling of PD neurodegeneration using the neurotoxins 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) and 1-methyl-4-phenyl-pyridinium (MPP(+)) has identified changes in gene expression of different endoplasmic reticulum (ER) stress proteins associated with MPTP- and PD-related neurodegeneration. We show that the protein disulfide isomerase (PDI) family member pancreatic protein disulfide isomerase (PDIp), previously considered exclusively expressed in pancreatic tissue, is uniquely upregulated among PDI family members within 24 h following exposure of retinoic acid (RA)-differentiated SH-SY5Y human neuroblastoma cells to either 1 mM MPP(+) or 10 microM of the highly specific proteasome inhibitor lactacystin. RT-PCR confirms PDIp expression in brain of post-mortem human PD subjects and immunohistochemical studies demonstrate PDIp immunoreactivity in LBs. Collectively, these findings suggest that increased PDIp expression in dopaminergic (DA) neurons might contribute to LB formation and neurodegeneration, and that this increased PDIp expression may be the result of proteasome impairment. PMID- 15353227 TI - Specific alteration of the expression of selected hypothalamic neuropeptides during acute and late mouse brain infection using a morbillivirus: relevance to the late-onset obesity? AB - Neurotropic viruses are involved in pathologies of the central nervous system, triggering transient or irreversible disorders, such as neurological diseases or homeostasis imbalance. In experimental animals, viruses have been shown to cause obesity, a complex disease depending on multiple factors, including genetic susceptibility and environmental components. Using a mouse model of virally induced obesity following brain infection by the Canine Distemper Virus (CDV), a morbillivirus closely related to the human measles virus, we investigated the modulation of expression of several hypothalamic neuropeptides known to intervene in the regulation of body weight and energy expenditure, both during the acute and late stages of infection. During the acute stage, while viral replication occurs, we found a dramatic decrease of expressions of neuropeptides, in particular neuropeptide Y, melanin-concentrating hormone (MCH), hypocretin, vasopressin and tachykinins, the magnitude of which seemed to be linked to the viral burden and the individual susceptibility. The effect of the virus, however, varied with the hypothalamic nucleus and neuropeptide involved, suggesting that certain circuits were affected while others remained intact. During the late stage of infection, marked recovery to the initial hypothalamic levels of peptide expression was seen in a number of lean animals, suggesting recovery of homeostasis equilibrium. Interestingly, some neuropeptidergic systems remained disturbed in mice exhibiting obese phenotype, arguing for their involvement in triggering/maintaining obesity. Even though our data could not fully explain the viral-induced obesity, they may be helpful in understanding the molecular events associated with obesity and in investigating therapeutic alternatives. PMID- 15353228 TI - Regional differences in the expression of Fos-like immunoreactivity after central salt loading in conscious rats: modulation by endogenous vasopressin and role of the area postrema. AB - In this study, we examined the quantitative relationship between centrally administered hypertonic saline (HS) concentrations and the expression of Fos-like immunoreactivity (FLI) in brain regions involved in the homeostasis of body fluids. The regions examined were the organum vasculosum laminae terminalis (OVLT), the median preoptic nucleus (MnPO), the subfornical organ (SFO), the paraventricular nucleus (PVN), the supraoptic nucleus of the hypothalamus, the nucleus of the solitary tract (NTS), and the area postrema (AP). The experiments were performed in conscious rats with attention to the actual changes in central [Na(+)]. Hypertonic saline (0.3, 0.67, or 1.0 M) was delivered at 1 microl/min for 20 min. The changes in cerebrospinal fluid [Na(+)] during i.c.v. administration of 0.3 M hypertonic saline were compatible with those expected for thermal dehydration. FLI increased in a dose-dependent manner in the dorsomedial cap of the PVN and NTS. Although the pressor responses during central salt loading were not significantly affected by pretreatment with the peripheral vasopressin V(1) receptor antagonist OPC-21268, FLI expression in the PVN was significantly augmented. In addition, in AP-lesioned rats, FLI expression in the lateral magnocellular part of the PVN and NTS was significantly enhanced after central salt loading. These results suggest that the peripheral vasopressin system participates in negative feedback to modulate neuronal activities in the PVN, probably through the AP or direct action at the PVN in response to central osmotic and/or Na(+) stimulation. PMID- 15353229 TI - Akt is activated via insulin/IGF-1 receptor in rat retina with episcleral vein cauterization. AB - The Akt serine/threonine kinase mediates pro-survival signalings in retina and was reported to be activated in a response to some retinal and optic nerve injuries. Human and experimental glaucoma induce apoptosis of retinal ganglion cells (RGCs). The purpose of this study is to test whether episcleral vein cauterization (EVC) to chronically elevate intraocular pressures (IOPs) in rats increase apoptosis of RGCs and affect activation of Akt and its upstream insulin like growth factor (IGF)-1 receptor/Insulin receptor. Three episcleral veins in left eyes of Sprague-Dawley rats were cauterized to elevate IOPs. Up to 6 months, IOPs were monitored and the retina was dissected at several time points. The numbers of terminal dUTP nick end labeling (TUNEL)-positive cells and those of RGCs labeled with fluorogold were counted in flat-mounted retina. Immunohistochemistry and immunoblotting were performed to identify cells expressing phosphorylated Akt and to quantify the phospho- to total ratios of Akt and IGF-1 receptor/insulin receptor. EVC significantly elevated IOPs up to 2 months, increased TUNEL-positive cells in an IOP-dependent fashion, and reduced 34.5% of RGCs at 6 months (P<0.001) compared with contralateral retinas. Phosphorylated Akt was specifically expressed in RGCs until 1 month after cauterization. Akt (P=0.036) and IGF-1 receptor/Insulin receptor (P=0.003) were transiently phosphorylated at 3 days. Intrinsic activation of the IGF-1 receptor/Insulin receptor to Akt pathway may occur in RGCs in retina with EVC. PMID- 15353230 TI - Evidence for biological effects of exogenous LPA on rat primary afferent and spinal cord neurons. AB - There is growing behavioural evidence that the phospholipid growth factor lysophosphatidic acid (LPA) modulates nociceptive responses in vivo. The present study investigated further the effects of LPA on peripheral nociceptive processing. Effects of intraplantar injection of LPA on ongoing and peripheral mechanically evoked responses of spinal neurons were studied in vivo. In addition, LPA-evoked responses of adult rat dorsal root ganglion (DRG) neurons were studied with calcium imaging. To determine whether LPA may also act at the level of the spinal cord, LPA receptor G-protein coupling in lumbar spinal cord sections was studied with in vitro autoradiography of guanylyl 5'-[g [(35)S]thio]triphosphate ([(35)S]GTPgammaS) binding. Intraplantar injection of LPA (5 microg/5 microl) significantly increased the duration (P<0.001) and frequency of spinal neuronal firing (P<0.01), compared to vehicle. Intraplantar injection of LPA (1 microg/5 microl) did not significantly alter innocuous and noxious mechanically evoked responses of spinal neurons, but a higher dose of LPA (5 microg) significantly (P<0.05) attenuated mechanically evoked responses of spinal neurons. Calcium imaging studies demonstrated that LPA (0.001-3 microM) increases intracellular calcium concentration in adult DRG neurons, suggesting that LPA can produce direct effects on. Incubation of spinal cord sections with LPA (1 microM) significantly (P<0.001) increased [(35)S]GTPgammaS binding in the superficial laminae of the dorsal horn of the spinal cord, suggesting that LPA may also have biological effects at this level. These data provide further evidence that exogenous LPA can modulate nociceptive processing and suggest that this may be mediated by a direct effect on primary afferent nociceptors. PMID- 15353231 TI - Regulators of G-protein signaling 4: modulation of 5-HT1A-mediated neurotransmitter release in vivo. AB - Regulators of G-protein signaling (RGS) play a key role in the signal transduction of G-protein-coupled receptors (GPCRs). Specifically, RGS proteins function as GTPase accelerating proteins (GAPs) to dampen or "negatively regulate" GPCR-mediated signaling. Our group recently showed that RGS4 effectively GAPs Galpha(i)-mediated signaling in CHO cells expressing the serotonin-1A (5-HT(1A)) receptor. However, whether a similar relationship exists in vivo has yet to be identified. In present studies, a replication-deficient herpes simplex virus (HSV) was used to elevate RGS4 mRNA in the rat dorsal raphe nuclei (DRN) while extracellular levels of 5-HT in the striatum were monitored by in vivo microdialysis. Initial experiments conducted with noninfected rats showed that acute administration of 8-OH-DPAT (0.01-0.3 mg/kg, subcutaneous [s.c.]) dose dependently decreased striatal levels of 5-HT, an effect postulated to result from activation of somatodendritic 5-HT(1A) autoreceptors in the DRN. In control rats receiving a single intra-DRN infusion of HSV-LacZ, 8-OH-DPAT (0.03 mg/kg, s.c.) decreased 5-HT levels to an extent similar to that observed in noninfected animals. Conversely, rats infected with HSV-RGS4 in the DRN showed a blunted neurochemical response to 8-OH-DPAT (0.03 mg/kg, s.c.); however, increasing the dose to 0.3 mg/kg reversed this effect. Together, these findings represent the first in vivo evidence demonstrating that RGS4 functions to GAP Galpha(i)-coupled receptors and suggest that drug discovery efforts targeting RGS proteins may represent a novel mechanism to manipulate 5-HT(1A)-mediated neurotransmitter release. PMID- 15353232 TI - Serotonergic modulation of psychological stress-induced alteration in synaptic plasticity in the rat hippocampal CA1 field. AB - In order to elucidate possible involvement of the serotonergic neuronal system in the stress-induced alteration in synaptic plasticity, the effects of contextual fear conditioning (CFC) on long-term potentiation (LTP) in the hippocampal CA1 field were examined in 5-HT-depleted rats by pretreatment with 5,7 dihydroxytryptamine (5,7-DHT, 200 microg/rat, i.c.v.). LTP induction was suppressed by footshock (FS) stimulation in 5-HT-lesioned rats and vehicle treated controls. When rats were exposed to CFC, which was received 24 h after FS stimulation, LTP was also blocked in both-treated groups. CFC-induced impairment of LTP, however, significantly attenuated in 5-HT-lesioned rats when compared with that in controls. Fear-related freezing behavior after FS stimulation occurred similarly in both treated groups, whereas the behavior observed during exposure to CFC significantly reduced in 5-HT-lesioned rats. These results suggest that the serotonergic mechanism is involved in the psychological stress induced alteration in synaptic plasticity, which appears to be associated with fear-related behavior. PMID- 15353233 TI - Timed hypocaloric food restriction alters the synthesis and expression of vasopressin and vasoactive intestinal peptide in the suprachiasmatic nucleus. AB - In mammals, the main circadian pacemaker is located in the suprachiasmatic nucleus (SCN) and its most potent synchronizer is the daily variation of the intensity of light. However, other nonphotic cues, such as timed food restriction, can induce changes in the circadian rhythms, leading also to the appearance of a food-entrained oscillator. The present study was designed to establish if the alterations of the circadian rhythms induced by timed hypocaloric food restriction are accompanied by structural changes in the SCN. Two groups of adult rats, both maintained on 12-h light/12-h dark cycles, were used; in one group, animals had permanent free access to food, whereas in the other they were subjected to a restricted hypocaloric early morning feeding during 7 months. Using stereological techniques and in situ hybridization, we have examined the structure of the SCN and the synthesis and expression of vasopressin (AVP) and vasoactive intestinal peptide (VIP). The volume of the SCN and the total number of neurons did not vary between the two groups. However, the total number of AVP- and VIP-immunoreactive neurons and the AVP and VIP mRNA levels were significantly decreased in timed hypocaloric food-restricted animals. The results indicate that timed hypocaloric food restriction has led to changes of AVP and VIP content of the neurons. They furthermore suggest the existence of a coupling between the food-entrainable oscillator and the light-entrainable pacemaker. PMID- 15353234 TI - Reduction of brain injury by antithrombotic agent acutobin after middle cerebral artery ischemia/reperfusion in the hyperglycemic rat. AB - In vivo magnetic resonance imaging (MRI) was used to observe the effect of acutobin, a purified thrombin-like enzyme (TLE), isolated from the snake venom of Deinagkistrodon acutus, on MRI-detected brain lesion volume and tissue perfusion deficit in a hyperglycemic rat right middle cerebral artery occlusion/reperfusion (MCAO/R) model. Acutobin (0.75 U/ml) was intravenously injected with a dosage of 2.5 U/kg body weight 30 min after MCAO (MCAO duration=60 min) and again 24 h after reperfusion. Multislice diffusion weighted imaging (DWI) and single-slice dynamic bolus tracking gradient echo (GE) imaging were sequentially acquired before and after MCAO/R. DWI-detected lesion volume was significantly (p<0.05) reduced by 24-31% from 350+/-45, 369+/-45 and 374+/-36 mm(3) in the saline treated group to 239+/-17, 282+/-26 and 259+/-32 mm(3) at 3, 4 and 24 h after reperfusion in the acutobin-treated group, respectively. Residual cerebral blood flow (CBF) in the right hemisphere recovered and remained at approximately 80% of normal perfusion over the measurement period in the acutobin-treated group, compared to approximately 40% in the saline-treated group. Mortality at 1 week after MCAO/R in the acutobin-treated group was significantly lower (25% mortality) than the saline control group (85% mortality). Our results indicate that acutobin improves brain tissue perfusion and reduces infarct volume and mortality in the hyperglycemic rat MCAO/R model. PMID- 15353235 TI - Degeneration of cholecystokinin-immunoreactive afferents to the VPL thalamus in a mouse model of Niemann-Pick disease type C. AB - Niemann-Pick disease type C (NP-C) is a progressive neurological disorder of lipid metabolism. The Balb/C npc1 mutant strain is a genetically authentic murine model of NPC, which reproduce the clinical and histologic features of human NP-C. In the present study, we show that cholecystokinin (CCK)-immunoreactive fibers in the thalamic VPL nuclei, which are densely distributed in controls, degenerate in NPC mice. This degeneration is associated with the appearance of CCK immunoreactive axonal spheroids containing characteristic intracellular inclusions of NP-C. These observations provide supportive evidence of the occurrence of dying-back axonopathy of neurons in the dorsal column nuclei in this mouse model. PMID- 15353236 TI - Effect of housing conditions on sulpiride-induced increases in extracellular dopamine (DA) levels in the nucleus accumbens of alcohol-preferring (P) rats. AB - The effect of housing conditions on sulpiride-induced increases in extracellular dopamine (DA) levels was investigated in the nucleus accumbens (NAC) of P rats. Rats were double-housed (DH) in plastic tubs, or single-housed (SH) in hanging wire cages for 12 weeks. Microdialysis in the NAC showed greater sulpiride induced DA increases in the NAC of SH vs. DH rats, with no difference in basal levels. The data indicate that housing conditions can alter DA D(2) receptor function in the NAC. PMID- 15353237 TI - Asymmetrically lesioned mesencephalon in healthy rodents: call for caution. AB - Stereological counting of tyrosine-hydroxylase immunoreactive (TH-IR) neurons in the mesencephalon is a pivotal parameter in assessing the extent of lesioning in animal models of Parkinson's disease. We here show that the number of TH-IR neurons often appears abnormally decreased in healthy--commercially available- mice and rats, although both the number of Nissl-stained cells and the striatal dopaminergic innervation are unaffected. This potential bias in assessing extent of neurotoxin-induced lesion and subsequent protection by pharmacological manipulation prompts us to call for caution in setting up experimental designs. PMID- 15353238 TI - The alpha7 nicotinic acetylcholine receptor-selective antagonist, methyllycaconitine, partially protects against beta-amyloid1-42 toxicity in primary neuron-enriched cultures. AB - Studies have suggested that the neuroprotective actions of alpha7 nicotinic agonists arise from activation of receptors and not from the extensive desensitization which rapidly follows activation. Here, we report that the alpha7 selective nicotinic antagonist, methyllycaconitine (MLA), protects against beta amyloid-induced neurotoxicity; whereas the alpha4beta2-selective antagonist, dihydro-beta-erythroidine, does not. These findings suggest that neuroprotective actions of alpha7-acting agents arise from receptor inhibition/desensitization and that alpha7 antagonists may be useful neuroprotective agents. PMID- 15353239 TI - Effect of the synthetic polyamine N,N'-bis-(3-aminopropyl) cyclohexane-1,4 diamine (DCD) on rat spinal cord nociceptive transmission. AB - In rats submitted to a C-fiber reflex response paradigm, intravenous (i.v.) administration of 2.5, 5 and 10 mg/kg of the synthetic polyamine N,N'-bis-(3 aminopropyl) cyclohexane-1,4-diamine (DCD) dose-dependently reduced both the integrated C reflex responses and wind-up activity. Inhibitory effects of the polyamine on spinal cord nociceptive transmission are likely to be consequence of blockade by extracellular DCD of NMDA receptor channels localized in dorsal horn neurons, although modulatory actions at supraspinal level and at other ion channels could also be possible. PMID- 15353240 TI - Validation of ion chromatography for human salivary anionic analysis. AB - OBJECTIVE: Assess the applicability of ion chromatography (IC) for human saliva studies. MATERIALS AND METHODS: The behaviour of an ion chromatograph (DX-100, Dionex) in the analysis of human saliva was examined in the following respects: calibration, reproducibility, recovery, and effect of pre-treatment conditions (dilution, temperature). The eluents used were 1.7 mM NaHCO3 + 1.8 mM Na2CO3 + 100 mg/L p-cyanophenol (2 mL/min) and 1.6 mM perfluorobutyric acid + 0.3 mM sodium hydroxide (0.3 mL/min) for inorganic and organic anions, respectively. RESULTS: The calibration curves were satisfactorily linear, but did not have unit slope (log-log plot) in the case of chloride, phosphate, sulphate and acetate. No significant effects were detected for centrifugation temperature, or between runs on the same occasion. CONCLUSION: IC appears to be a valuable technique well suited to the efficient analysis of several salivary anions. PMID- 15353241 TI - Human salivary anionic analysis using ion chromatography. AB - OBJECTIVE: To survey concentrations and time-variability of the major anions of human saliva using ion chromatography (IC). MATERIALS AND METHODS: Resting saliva samples after overnight fasting, obtained under standardized conditions in time series for 10 subjects, were analysed for chloride, phosphate, nitrate, sulphate, thiocyanate, acetate and lactate. A second group of five subjects provided mixed saliva as before, and in addition a sample of parotid saliva collected immediately afterwards using a Lashley cup. Analysis was by ion chromatography (DX-100, Dionex) using 1.7 mM NaHCO3 + 1.8 mM Na2CO3 + 100 mg/L p-cyanophenol (2 mL/min) and 1.6 mM perfluorobutyric acid + 0.3 mM sodium hydroxide (0.3 mL/min) as eluents for inorganic and organic anions, respectively. RESULTS: The time series data showed great natural variability. Analysis of the correlations showed that the concentration of nitrate was uncoupled from those of the other anions. Lactate was not detected. Parotid saliva showed marked differences from mixed saliva, most notably the absence of acetate and the presence in two instances of lactate. Bromide, nitrite and citrate were not detected. CONCLUSIONS: Acetate is a major component of the oral chemical environment. Simultaneous analyses permit the investigation of the relationships between component concentrations that otherwise might have been difficult to study. PMID- 15353242 TI - Processing of acidic proline-rich proprotein by human salivary gland convertase. AB - Previously it was found that proproteins for basic and glycosylated salivary proline-rich proteins (PRP) were cleaved prior to secretion from cells by furin, a well-known convertase. In contrast proproteins for acidic PRPs are not cleaved by furin or other convertases. To investigate the convertase responsible for in vivo processing of acidic PRP proproteins, homogenates of human sublingual glands were fractionated by centrifugation at 10,000 x g and 100,000 x g and activity demonstrated in all fractions. The 100,000 x g pellet was fractionated into Golgi, smooth endoplasmic reticulum and microsomal fractions with the latter containing the enzyme. Subfractionation of the microsomes revealed that the activity was located in the membrane proteins. Since the microsomes contain components of the secretory pathway the enzyme in this fraction may be responsible for intracellular cleavage of the acidic PRP proprotein. The enzyme was active at alkaline pH. It was strongly inhibited by metal chelators indicating that it is a metalloprotease. It was not inhibited by an acid protease inhibitor, but partly inhibited by some serine protease inhibitors indicating that serine proteases may play a role in degradation. Co2+ and to some extent Zn2+ activated the enzyme, but it was strongly inhibited by Hg2+ and Cu2+ as well as the organomercurial p-chloromercuribenzenesulfonic acid. Thus it appears that the enzyme contains an important -SH group. These characteristics indicate that the convertase is related to a group of metal- and thiol-dependent proteases known as thimet oligopeptidases, but in contrast to the latter enzymes the sublingual convertase was not inhibited by angiotensin antagonists. PMID- 15353243 TI - Tissue distribution and nucleotide sequence of bovine mRNA for salivary proline rich protein P-B. AB - The tissue distribution of P-B was investigated to obtain information on the physiological significance of this proline-rich protein. To design primers and probes for a tissue distribution analysis, a polymerase chain reaction (PCR) based cloning of bovine P-B cDNA was performed using tooth germ and the nucleotide sequence was determined. The cloned bovine P-B cDNA was composed of 356 bp and included the region corresponding to the mature P-B protein and part of the 3' non-coding sequence. This part of the sequence is identical to the corresponding region of human P-B cDNA from the submaxillary gland. DNA corresponding to the P-B mRNA was amplified by PCR using cDNAs from various bovine tissues including tooth germ, submaxillary gland, parotid gland, lachrymal gland, heart, liver, stomach, pancreas, spleen, kidney, adrenal, and ovary. A quantitative analysis indicated the heart, submaxillary gland, tooth germ and kidney to be major sites of P-B expression. The ubiquitous distribution of P-B mRNA among bovine tissues together with findings of the presence of genes hybridizable with a DNA probe for P-B among species such as human, bovine, rat, mouse, and yeast as reported previously suggested a fundamental physiological role for this protein. PMID- 15353244 TI - Nitric oxide-dependent mitotic activity in salivary glands of the rat upon sympathetic stimulation. AB - Incorporation of [3H]thymidine into trichloroacetic acid (TCA)-insoluble material of the parotid and submandibular glands was used as an index of mitotic activity following unilateral electrical stimulation of the sympathetic innervation (20 Hz, 4 min every fifth minute over 34 min). Stimulation under beta-adrenoceptor blockade (propranolol 2 mg/kg, intravenous) alone or combined with alpha adrenoceptor blockade (phentolamine 2 mg/kg, intravenous) did not increase the rate of [3H]thymidine incorporation into the two types of glands. However, under alpha-adrenoceptor blockade the [3H]thymidine incorporation increased into the parotid glands, by 122% (compared to the glands on the contralateral side), but not into the submandibular glands. In the presence of the neuronal type NO synthase (nNOS) blocker N-PLA (30 mg/kg, intravenous) or the unselective NO synthase blocker L-NAME (30 mg/kg, intravenous), this increase was reduced to 49 and 47%, respectively. Thus, the major part of the sympathetically nerve-evoked beta-adrenoceptor-mediated mitotic response was found to depend on the activity of neuronal type NO-synthase to generate NO. Since the sympathetic nerve fibres of the parotid gland lack NO-synthase, the neuronal type NO-synthase subjected to the inhibitors is likely to be of parenchymal origin. PMID- 15353245 TI - Evaluation of acetaminophen P-glycoprotein-mediated salivary secretion by rat submandibular glands. AB - The constant ratio between saliva and plasma acetaminophen concentrations (S/P) during the elimination phase is assumed to result from the equilibrium established among the free-drug concentrations in the arterial blood, venous blood and saliva. Salivary secretion of acetaminophen is assumed to result from a passive diffusion of the drug to saliva from the blood that supplies the salivary glands. However, the constant S/P ratio during acetaminophen disposition and the finding that P-glycoprotein (P-gp), a protein recognized to pump substrates out of the cell, is expressed in duct cells of the submandibular glands questions the mechanisms involved in acetaminophen salivary secretion. Thus, we intended to evaluate the existence of a P-glycoprotein-mediated transport of acetaminophen in rat submandibular glands. Acetaminophen (30 mg/kg, i.v.) pharmacokinetics was assessed in controls and in rats pre-treated with erythromycin (100 mg/kg) as a P glycoprotein inhibitor. Acetaminophen pharmacokinetic parameters were calculated from saliva and plasma levels considering a non-compartmental analysis. Mean plasma and salivary profiles of control and pre-treated animals were almost superimposable. No difference could be found in S/P ratios in control and erythromycin pre-treated animals (P > 0.05). Moreover, no statistical difference could be found in the kinetic parameters calculated from saliva or plasma drug level (P > 0.05). These observations indicate that acetaminophen salivary secretion in rat submandibular glands is not related to P-glycoprotein-mediated transport under the experimental conditions of the present work. PMID- 15353246 TI - Protective effect of the in situ formed short-term salivary pellicle. AB - Salivary pellicle, as previously investigated, protects the enamel surface after certain processes of maturation against the influence of acidic agents. The aim of the present study was to investigate the protective effect of the short-term salivary pellicle formed in situ over periods of 3, 60 and 120 min. Six human volunteers used intraoral acrylic splints with bovine enamel samples fixed at the buccal and palatal sites of the maxillary first molars and second premolars. Enamel specimens (n = 252) with and without pellicle were immersed for 60 s in 1.0% citric acid solution under agitation. Knoop surface hardness (KHN) of uneroded polished enamel was measured as a baseline and estimated immediately after erosive treatment reflecting the microhardness loss (DeltaKHN). The amounts of calcium dissolved from the eroded enamel surface were analysed by atomic absorption spectroscopy and scored in mg/l per 10 mm2 of enamel surface area. In addition, the scanning electron microscope was used for the micromorphological examination of the erosive alterations of the enamel surface. The average microhardness loss values after erosion of the enamel samples with buccally/palatally formed pellicle layers were measured as 139.1/144.9 DeltaKHN for 3 min pellicle, 145.9/146.9 DeltaKHN for 60 min pellicle and 141.7/138.6 DeltaKHN for 120 min pellicle. Calcium release values from the specimens with buccal/palatal pellicles were amounted to 15.0/14.9, 16.5/15.9 and 15.3/17.4 mg/l per 10 mm2 for 3, 60 and 120 min-old pellicles, respectively. No significant differences were related to the pellicle formation time and intraoral site (buccal or palatal) in all tested series (ANOVA, P < 0.05). However, significant protection of the enamel surface provided by the pellicle layer was observed on all pellicle-covered surfaces if compared to the non-covered enamel samples (calcium release: 25.6 mg/l per 10 mm2; microhardness loss 187.0 DeltaKHN). These data were in accordance with the morphologic alterations caused by citric acid on the pellicle-covered and pellicle non-covered specimens. It could be concluded that salivary pellicle formed in situ within a period of 3 min offers protection of enamel against citric acid. However, pellicle does not completely inhibit the erosive action of citric acid under the conditions of the present study. PMID- 15353247 TI - TGF-beta activated Smad signalling leads to a Smad3-mediated down-regulation of DSPP in an odontoblast cell line. AB - OBJECTIVE: Transforming growth factor-beta (TGF-beta) regulates odontoblast differentiation and stimulates dentine extracellular matrix synthesis. However, until recently, the molecular mechanisms of action of TGF-beta have been unknown. Smad proteins have recently been identified as intracellular signalling mediators of TGF-beta. In this study, we characterise the role of Smad proteins as mediators of TGF-beta in a mouse odontoblast cell line MDPC-23. METHODS: Transcription of Smads was detected by RT-PCR. The change of intracellular location of Smad proteins treated by TGF-beta1 was evaluated immunocytochemically. Smad function and its role in transcription of dentin sialophosphoprotein (DSPP) were investigated in cotransfection experiments using promoter-luciferase reporter gene constructs. RESULTS: MDPC-23 cells expressed Smad2, Smad3 and Smad4 mRNA. Endogenous Smad2, Smad3 and Smad4 rapidly translocated from the cytoplasm into the nucleus in response to TGF-beta1. The activity of the TGF-beta-responsive p3TP-Lux reporter construct was stimulated by 12.7-fold with TGF-beta1 treatment. Over-expression of wild-type Smad3 promoted TGF-beta1-induced luciferase activity, whereas dominant negative Smad3 inhibited it. TGF-beta1 also inhibited the activity of DSPP promoter luciferase reporter construct containing the sequence between -791 bp and +54 bp of the mouse DSPP gene. Over-expression of wild-type Smad3 potentiate the inhibitory effect of TGF beta1 on transcriptional regulation of DSPP, while dominant negative Smad3 decreased the effect. In contrast to Smad3, wild-type Smad2 or its dominant negative mutant had little effect on TGF-beta1 regulation of the promoter activity of DSPP. CONCLUSIONS: Smad2, Smad3 and Smad4 are present and activated by TGF-beta1 in MDPC-23 cells. The Smad pathway is functional in these cells and Smad3 appears to be involved in down-regulation of DSPP by TGF-beta1. These findings raise the possibility that Smad signalling plays a role in dentinogenesis. PMID- 15353248 TI - Radiodensity of enamel and dentin of human, bovine and swine teeth. AB - Several studies have aimed to evaluate the characteristics of hard dental tissues from animal species in order to adequately substitute the usage of human teeth. The purpose of this study was to evaluate the radiodensity of enamel and dentin of human, bovine and swine teeth. Five specimens of 2 mm in thickness were obtained from human, bovine and swine teeth and the radiographic images were taken positioning it on a phosphor plaque digital system, Digora (Soredex, Helsinki, Finland). The radiodensity of each specimen was obtained and data were compared by ANOVA following Tukey test (P < 0.05). The results showed that human and bovine enamel presented similar radiodensity, which was higher than the one from swine enamel; bovine and swine dentin presented similar radiodensity, and only bovine dentin presented greater similarity to human dentin. Bovine teeth seems to be more similar to human teeth in respect to radiodensity. PMID- 15353249 TI - Crystalline structure of dental enamel after Ho:YLF laser irradiation. AB - Irradiation of teeth with lasers using specific wavelengths and energy densities produces surface melting. This effect has been already applied to different procedures such as caries prevention and hypersensitivity reduction. The aim of this study is to characterize the crystalline structure of bovine enamel after holmium laser irradiation. A holmium laser (Ho:YLF) with emission wavelength of 2065 nm was used. Enamel tissues were irradiated in ablative regime and their structures before and after irradiation were analyzed using the powder X-ray diffraction technique. The X-ray diffraction patterns of non-irradiated enamel correspond to carbonated hydroxyapatite and those produced by irradiated samples indicate the existence of a mixture of two crystalline phases: hydroxyapatite and tetracalcium phosphate. The structural characteristics of holmium irradiated enamel were compared with those of the same tissue irradiated with other lasers. PMID- 15353250 TI - Quantitative analysis of the impact of the organic matrix on the fluoride effect on erosion progression in human dentine using longitudinal microradiography. AB - After an initial demineralisation, an intensive fluoridation is capable of inhibiting the erosive mineral loss in dentine completely, which might be related to the presence of the exposed organic dentine matrix. Aim of the present study was, therefore, to evaluate whether fluoride is also effective when the organic material is removed. The study was a cyclic de- and remineralisation model over 5 days in vitro. Samples from human coronal dentine were demineralised with citric acid (pH 2.3; 6 x 10 min per day) and intermittently stored in a remineralisation solution. Groups (n = 25 each) were defined as follows: Group 1: erosion only, no fluoridation; Group 2: erosion, alternately fluoridation with toothpaste (NaF; 0.15% F-; 3 x 5 min per day), mouthrinse (Olaflur/SnF2; 0.025 F-; 3 x 5 min per day) and gel (Olaflur/NaF; 1.25% F-; at Days 1 and 3 instead of one toothpaste application); Group 3: erosion and fluoridation as Group 2, organic material was continuously removed with collagenase (from Clostridium Histolyticum type VII, 100 U/ml) added to the remineralisation solution. Mineral content was monitored daily using longitudinal microradiogaphy. After fluoridation in the presence of collagenase, a linear increase in mineral loss (73.3 +/- 17.6 microm at Day 5) was observed, which significantly (P < or = 0.001) exceeded that of the control group (45.9 +/- 14.3 microm at Day 5). After fluoridation in the absence of collagenase, mineral loss ceased after the 2nd day (12.2 +/- 10.2 microm at Day 5) and was significantly lower compared to Groups 1 and 3 (P < or = 0.001). The results indicate that the dentine matrix is essential for the effectiveness of fluoride in dental erosion. PMID- 15353251 TI - X-ray microtomographic study of mineral concentration distribution in deciduous enamel. AB - OBJECTIVE: To determine the mineral concentration distribution in deciduous enamel by quantitative X-ray microtomography (XMT). DESIGN: Tooth rods ( approximately 2 mm x 2 mm) were removed from the mid-buccal region of 11 deciduous molars. Three XMT slices were taken at 1.5, 2.0 and 2.5 mm from the amelocemental junction. The distribution and variation in mineral concentration of enamel were studied from the XMT images. RESULTS: The mean mineral concentration for all the teeth was 2.81 (S.D. = 0.065) g cm(-3). There was no notable difference in the mean mineral concentration values between the three XMT slices of each tooth. However, there was up to 8% variation between different teeth (2.69-2.92 g cm(-3)). Gradients of increasing mineral concentration from the amelodentinal junction (ADJ) to the external surface were found, ranging from 0.08 to 0.60 mg cm(-3) microm(-1) with a mean of 0.366 mg cm(-3) microm(-1). The mineral concentration gradients in the occlusal slices were steeper than those in the cervical slices. The difference in mineral concentration between the inner and outer enamel ranged from 1.5 to 8.7%. CONCLUSION: In view of the large variation in both the means and the gradients of mineral concentration in deciduous molars, the mineral distribution of each experimental tooth should be measured as baseline data in studies of caries progression. PMID- 15353252 TI - Expression of growth factors in the gingival crevice fluid of patients with phenytoin-induced gingival enlargement. AB - The mechanism underlying phenytoin (PHT)-induced gingival enlargement (GE) is not yet known. The aim of the present study was to investigate transforming growth factor-beta1 (TGF-beta1), platelet-derived growth factor-BB (PDGF-BB) and basic fibroblast growth factor (bFGF) profiles in the gingival crevice fluid (GCF) of patients with PHT-induced GE and to compare the results with healthy controls. Five PHT-treated patients and five healthy subjects with normal periodontal tissue were included in this study. GCF samples were collected from (i) enlarged gingival sites in patients receiving PHT (GE+); (ii) non-enlarged gingival sites in the same patients (GE-); (iii) normal gingival sites of healthy subjects (control). The levels of TGF-beta1, PDGF-BB and bFGF in the GCF samples were analysed by ELISA. The results showed that the total amounts of TGF-beta1 and PDGF-BB in the GE+ group were higher than in the GE- group and significantly higher than in the control group (P < 0.05). However, no significant differences were found between the groups when the concentrations of these growth factors were compared. bFGF levels were not compared as this growth factor could be detected in only 33, 41 and 44% of the GE+, GE- and control GCF samples, respectively. These results show that TGF-beta1 and PDGF-BB are readily detectable in GCF obtained from enlarged and non-enlarged sites of PHT recipients and suggest that since the amounts were markedly higher at the GE+ than the GE- sites, the systemic administration of PHT has a pronounced localised effect on the levels of these growth factors. Moreover, our findings provide evidence that both TGF-beta1 and PDGF-BB are closely associated with the clinical manifestation of PHT-induced GE. PMID- 15353265 TI - Tachykinin-modulated anti-viral responses. AB - Following viral infection, the expression of substance P and its receptor can contribute significantly to the resulting host response. For gammaherpesvirus infection of mice, the presence of this tachykinin and its receptor contributes to the protective host response. It is likely that this augmentation of the immune response is directed toward the developing T helper type 1 response and cytotoxic T lymphocyte activation. However it has also been shown that the presence of substance P and its receptor may contribute to viral diseases by facilitating viral replication or by contributing to a destructive inflammatory response. Specifically, the presence of substance P can augment replication of HIV in cultured macrophages, which is especially significant since levels of this tachykinin are elevated in patients with this viral disease. Furthermore, rodent models of paramyxovirus infection have demonstrated that the presence of neurokinin receptors and their ligands contributes to the destructive inflammatory response in airways. Taken together, these studies demonstrate a surprising role for substance P and the neurokinin-1 receptor in the host response following these viral infections. PMID- 15353266 TI - Osmoadaptation and osmoregulation in archaea: update 2004. AB - The response of archaea to changes in external NaCl is reviewed and compared to what is known about osmoadaptation and osmoregulation in bacteria and eukaryotes. Cells placed in altered external NaCl exhibit short term and long term responses. The earliest events are likely to be water movement through aquaporin-like channels (efflux if external NaCl has been increased, influx into the cell if the external NaCl has been decreased) and ion movement (e.g., K+ moving in the direction opposite to water flow) through channels sensitive to osmotic pressure. A brief discussion of recent structures of homologues of these membrane proteins is presented. Accumulation of organic solutes, either by uptake from the medium or de novo synthesis, is triggered after these initial changes. Archaea have some unique organic solutes (osmolytes) that are not used by other organisms. These as well as other more common solutes have a role in stabilizing macromolecules from denaturation. Many osmolytes are distinguished by their stability in the cell and their lack of strong interactions with cellular components. A cell may respond by accumulating one or more temporary osmolytes, then over time readjust the intracellular solute distribution to what is optimal for cell growth under the new conditions. Coupled with the movement and accumulation of solutes is the induction of stress proteins (e.g., chaperonins) and, in some cases, transcriptional regulation of key enzymes. The response to NaCl stress of Methanococcus thermolithotrophicus is presented as an example of how one particular archaeon responds and adapts to altered osmotic pressure. The detailed response of many other archaea to osmotic stress will be needed in order to identify features (aside from some of the organic osmolytes) unique to the organisms in this kingdom. PMID- 15353267 TI - Cloning and sequencing of a cDNA encoding for a protein specifically expressed in human testis, ovary and placenta. AB - Reproductive cell/tissue-specific antigens are attractive candidates for the development of a contraceptive vaccine. Using the differential display technology, a human cDNA fragment of 322-bp, designated as T17, was identified showing specific expression in the human testis. The T17 cDNA fragment was used as a probe to screen the human testis cDNA-lambda gt10 library. After screening, one positive clone of approximately 1.1 kb having T17 nucleotide (nt) sequence was obtained. The 5' and 3' termini of this cDNA clone were extended by using 5' and 3' RACE procedures that yielded a full-length cDNA, designated as the TOP gene. The TOP cDNA is 1,480-bp long and has an ORF of 463 aa with the first ATG Met start codon at nucleotide (nt) 64 and the stop codon TGA at nt 1452. The translated protein has a calculated molecular mass of 49.3 kD with isoelectric point of 12.17. The deduced amino acid (aa) has one potential N-glycosylation site, and several phosphorylation and myristoylation sites. Hydropathy plot generated from the deduced aa sequence showed it to be a surface protein. Comprehensive computer search in the database did not reveal any homology to any existing sequences both at the nt and aa levels. The TOP cDNA was found to be completely localized on the human chromosome 16 at the nt position 507833-506354, with the TOP ORF at the nt position 507770-506354. Northern blot analysis using three human Northern blots, indicated the specific expression of TOP gene in the human testis, ovary and placenta. RT-PCR-Southern blot analysis also confirmed tissue-specific expression of the TOP gene. The TOP cDNA may help us to gain insight into transcriptional control of the differentially expressed reproductive tissue-specific genes. It may also find clinical applications in the development of a contraceptive vaccine, and specific diagnosis and treatment of infertility in humans. PMID- 15353268 TI - Oxygen free radical and antioxidant defense mechanism in cancer. AB - The reactive oxygen species (ROS) can damage the nucleic acids. The oxidative modification of the DNA constitutes the fundamental molecular event in carcinogenesis and that is why the interest in the study of the involvement of ROS in that process. On the other hand, oxidative DNA damage-induced mutagenesis is widely hypothesized to be a frequent event in the normal human cell. The enormous evidence suggests an important role of ROS in the expansion and progression of tumor clones, being considered a relevant class of carcinogens. In addition, the use of immunohistochemical techniques has showed that the various types of cancer examined to date manifest an imbalance in their antioxidant mechanisms to respect the primary cell. In the near future new insights in cancer therapies, based on modulation of cellular redox status, may lead the way to additional tools against carcinogenesis from ROS. PMID- 15353269 TI - Building a functional artery: issues from the perspective of mechanics. AB - Despite the many successes of arterial tissue engineering, clinically viable implants may be a decade or more away. Fortunately, there is much more that we can learn from native vessels with regard to designing for optimal structure, function, and properties. Herein, we examine recent observations in vascular biology from the perspective of nonlinear mechanics. Moreover, we use a constrained mixture model to study potential contributions of individual wall constituents. In both cases, the unique biological and mechanical roles of elastin come to the forefront, especially its role in generating and modulating residual stress within the wall, which appears to be key to multiple growth and remodeling responses. PMID- 15353270 TI - Regulation of magnesium content during proliferation of mammary epithelial cells (HC-11). AB - To study the role of Magnesium in the regulation of cell proliferation we characterized the proliferation behaviour of HC-11 mammary epithelial cells that were grown in media containing low to high Mg concentrations. Cells grown under control conditions (0.5 mM Mg in the medium) or in the presence of high (H) Mg (45 mM) displayed similar log-phases and reached confluence in 72h. In the presence of low (L) Mg (0.025 mM) the cells exhibited a reduced growth rate and did not reach confluence at 72h. Intra cellular total Mg increased from 12 to 36h of culture in all cells examined but returned to basal levels in those cells which reached confluence (i.e., control and H-Mg cells). Intra cellular Mg increased independent of mitosis-induced changes of volume and adenine nucleotides pools but correlated with an increased percentage of cells in the S phase and with total nucleic acid contents. These bell-shaped changes of intra cellular Mg were less evident in L-Mg cells, likely due to a combination of low Mg levels in the medium and decreased growth rate. Changes in membrane potential and pH were important factors that contributed to maintaining intra cellular Mg at physiologic levels in the face of increased or decreased availability of extra cellular Mg. H-Mg cells were depolarised and more acidic than control cells; conversely, L-Mg cells showed a pattern of hyperpolarization and alkalinization. These results lend support to the concept that Mg may be involved in regulating cell proliferation, and show that cells maintain adequate levels of intra cellular Mg, and hence their proliferation potential, even under conditions of extreme changes of extra cellular Mg. PMID- 15353271 TI - Expression of non-neuronal acetylcholine in lymphocytes and its contribution to the regulation of immune function. AB - Lymphocytes express most components of the cholinergic system including acetylcholine (ACh), muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively), choline acetyltransferase (ChAT), high affinity choline transporter and acetylcholinesterase. ACh and mAChR agonists elicit intracellular Ca2+ signaling, up-regulation of c-fos expression and nitric oxide synthesis within T and B cells probably via M3 and M5 mAChRs. Stimulation of nAChRs with ACh or nicotine causes a rapid and transient Ca2+ signaling in T and B cells, probably via alpha7 nAChR subunit-mediated pathways. Phytohemagglutinin- or antigen-induced T cell activation via cell surface molecules (e.g., T cell receptor/CD3 complexes) enhances lymphocytic cholinergic transmission by up regulating ChAT and M5 mAChR expression. It is thus likely that a local lymphocytic cholinergic system is involved in regulating immune function. This idea is supported by the findings that lymphocytic cholinergic activity is altered in animal models exhibiting immunological abnormalities. In addition, it appears likely that during interactions mediated by cell surface molecules T cells communicate via ACh with thymic epithelial cells and vascular endothelial cells, which also express ChAT and nAChRs or mAChRs. This interaction leads to T cell selection and maturation in the thymus and local vascular smooth muscle relaxation. Collectively, these data provide a compelling picture in which lymphocytes constitute a cholinergic system that is independent of cholinergic nerves, and which is involved in the regulation of immune function and local circulation. PMID- 15353272 TI - Laboratory diagnostic aspects of drug resistant tuberculosis. AB - Multi-drug resistant strains of Mycobacterium tuberculosis (MDR-TB) are present world wide, and in many areas constitute a serious threat to the efficacy of TB control programs. The most effective strategies to limit further spread of MDR-TB are rapid detection of drug resistance followed by prompt and effective therapy. Routine laboratory diagnosis of drug resistance in TB requires a viable, pure culture of M. tuberculosis. Use of liquid media has decreased the turn around time for susceptibility test results however, because of the slow growth of M. tuberculosis these assays can still take 10 to 14 days. Alternatively, an increased understanding of the molecular basis for resistance to the antituberculosis drugs can greatly contribute to further decreasing turn around time. Based on this information, more precise and rapid molecular testing can be developed and lead to more appropriate and timely treatment regimens. In this review, we discuss methods for, and problems encountered in, performing TB drug susceptibility assays. Descriptions of routine protocols will be followed by recent developments in molecular detection of drug resistance. PMID- 15353273 TI - The involvement of oxidative stress in bovine herpesvirus type 4-mediated apoptosis. AB - Bovine herpesvirus type 4 (BHV-4) belongs to the gamma-2-herpesviruses of the Gammaherpesvirinae subfamily. BHV-4 has a worldwide distribution and has been isolated in a variety of clinical diseases as well as from healthy cattle. In this report we demonstrate that BHV-4 induces apoptosis in MDBK cells. In the early phases of apoptosis, cells show an increase in the intracellular level of reactive oxygen species, which is indicative of oxidative stress. This precedes DNA fragmentation, a hallmark typical of apoptosis. Cells were protected from apoptosis only by certain antioxidants (butylated hydroxyanisole and ebselen), whereas N-acetylcysteine turned out to be ineffective. Antioxidants that protected cells from apoptosis prevented oxidative stress but failed to block virus growth. These observations suggest that oxidative stress may be a crucial event in the sequence leading to apoptotic cell death but apoptosis is not required for the multiplication of BHV-4. PMID- 15353274 TI - DNA replication licensing. AB - The DNA replication licensing system ensures that chromosomal DNA is replicated precisely once before cell division occurs. A DNA helicase must be loaded on origin DNA for replication to initiate. Considerable evidence suggests that the MCM complex acts as a replicative helicase in eukaryotes. When the MCM complex is loaded on the chromatin, the replication origin is formally defined as being licensed for replication. Licensing takes place several hours before origins are activated to undergo replication in S-phase. Genetic and biochemical studies show that the licensing process is well conserved in eukaryotes. Cyclin Dependent Kinases (CDKs), the master regulators of the cell cycle, coordinate the initiation of the two key cell cycle events, replication of DNA and its segregation at mitosis. Eukaryotes have developed complex regulatory mechanisms to ensure that origin licensing is coordinated with these events so that genome integrity is preserved during successive cell divisions. PMID- 15353275 TI - Eucaryotic replication origin binding proteins. AB - Initiation of eukaryotic DNA replication is a tightly controlled process. Replication initiates at multiple specific sites (replication origins) that have been licensed for replication, following the cell cycle-dependent, multi-step assembly of specific factors. Thus, replication origins occur in two chromatin states: a replication-competent pre-replicative (pre-RC) state, when a number of replication proteins assemble on the origin in a stepwise fashion, and a replication-incompetent post-replicative (post-RC) state, in which the origin (or elements of it) is bound only by the origin recognition complex (ORC) (or subunits of it). This review summarizes the origin binding proteins that have been have been identified to date. PMID- 15353276 TI - Demographic analysis and modeling of human populations exposed to ionizing radiation. AB - The health effects of ionizing radiation on human populations are often analyzed using epidemiological statistical methods. Because of the complexity of the health consequences of ionizing radiation and the prolonged period during which the consequences emerge, we propose to evaluate these health effects using mathematical models that are based on the best theoretical reasoning and prior biological evidence about disease mechanisms. We believe this will improve the ability of the model to identify health effects and reduce erroneous inferences. PMID- 15353277 TI - The role of tachykinins in central nervous system inflammatory responses. AB - While glial cells are recognized for their roles in maintaining neuronal function, there is growing appreciation of the ability of resident glial cells to initiate and/or augment inflammation following trauma or infection in the central nervous system (CNS). The tachykinin, substance P, is found throughout the CNS, with evidence for both neuronal and glial cells as being sources of this neuropeptide. Substance P is well known to augment inflammatory responses at peripheral sites, such as the gastrointestinal tract and skin, which raises the possibility that this tachykinin might serve a similar function within the brain. This review focuses on the evidence for tachykinins in regulating the immune functions of CNS glial cells. Neurokinin-1 (NK-1) receptors have a high affinity for substance P and are expressed by a number of resident CNS cell types, including astrocytes and microglia. Importantly, substance P/NK-1 receptor interactions elicit activation of signal transduction pathways in both cell types and can initiate, or augment, inflammatory responses by astrocytes and microglia. The ability of substance P to augment immune responses of glial cells has important ramifications for the development of protective host responses within the CNS or, alternatively, the progression of damaging inflammation. PMID- 15353278 TI - Non-neuronal mammalian tachykinin expression. AB - Mammalian tachykinins are traditionally viewed as neuropeptides. This review describes the mammalian tachykinins and evidence for expression of these peptides by non-neuronal cells. Tachykinin expression is defined as evidence for gene transcription, peptide production, or peptide secretion. Since the functions of mammalian tachykinins have been amply reviewed, the biological roles of these peptides will be noted briefly, with emphasis on immune cell action. Of particular interest is the predicted existence and non-neuronal expression of new mammalian tachykinins--hemokinin 1, the endokinins and C14TKL-1. Synthetic forms of these peptides have high affinity for the NK1 receptor, the protein traditionally associated with substance P binding. By acting on the same "substance P" receptor, these tachykinins have the potential for promoting similar post-receptor functions. The structure and action of representative non mammalian tachykinins acting on mammals are also presented. These peptides, of interest in their own right, also appear to exhibit selectivity for the NK1 receptor. They strengthen the notion that multiple ligands may be capable of binding to one receptor, NK1, effecting similar cellular responses. PMID- 15353279 TI - Cell spreading controls balance of prestress by microtubules and extracellular matrix. AB - The controversy surrounds the cellular tensegrity model. Some suggest that microtubules (MTs) must bear a significant portion of cell contractile stress (prestress) if tensegrity is a useful model. Previously we have shown that for highly spread airway smooth muscle cells (areas>2500 microm2) MTs balance a significant but small potion (average 14%) of the prestress. To further explore if controlling the degree of cell spreading could modulate the portion of the prestress balanced by MTs, we utilized a recent method by which tractions are quantified in cells that are constrained within micropatterned adhesive islands of defined sizes on the surface of flexible polyacrylamide gels containing fluorescent microbeads. The prediction is that if MTs balance a portion of the contractile stress, then, upon their disruption, the portion of the stress balanced by MTs would shift to the substrate, causing an increase in traction and strain energy. We first activated the cells maximally with histamine and then disrupted the MTs with colchicine. Histamine resulted in an increase in intracellular calcium whereas ensuing colchicine addition in the presence of histamine did not change intracellular calcium concentration, suggesting there was no additional net increase in contractile stress inside the cell. We found that following disruption of MTs the increase in traction and strain energy varied with the degree of cell spreading: as the cell projected areas increased from 500 micrometer 2 to about 1800 micrometer 2, the percent increase in tractions decreased from 80% to about a few percent and the percent increase in strain energy decreased from 200% to almost zero percent, indicating the portion of the prestress balanced by MTs decreased as the cells increased spreading. These findings demonstrate that complementary role of the extracellular matrix and the MTs in balancing the prestress is controlled by the degree of cell spreading. PMID- 15353280 TI - Quantifying cell-adhesion strength with micropipette manipulation: principle and application. AB - Quantifying cell-adhesion strength is of great importance in biology and medicine. Cell-adhesion strength can be characterized by separating two adherent cells and determining the force required to do so, or by measuring the lifetime of a receptor-ligand bond that mediates cell adhesion. To this end, several micropipette-based experimental techniques that operate at both cellular and molecular levels have been developed over the past few decades. In this review, we provide an overview of three of these techniques, i.e., the step-pressure technique (SPT), the biomembrane-force probe (BFP), and the micropipette aspiration technique (MAT). More detailed discussion will be given about the requirements and applications of the MAT. PMID- 15353281 TI - Anti-interferon antibodies in multiple sclerosis. Molecular basis and their impact on clinical efficacy. AB - Low levels of naturally-occurring, high-affinity antibodies directed against cytokines can be found in the circulation of individuals who have never been exposed to exogenously-supplied cytokines. These antibodies are thought to play a regulatory role in the intensity and duration of immune response. Interferon (IFN) beta has been shown to attenuate both relapsing-remitting multiple sclerosis (MS) and secondary progressive MS in several well-powered, randomized, controlled clinical trials. IFN therapy can induce the production of anti-IFN neutralizing antibodies (NAb), usually in the second 6 months of treatment, in 3 to 45% of treated patients. This variation in the proportion of NAb-positive patients is probably due to the immunogenicity of different formulations of IFN beta, as well as the assay used, which are not currently standardized. The occurrence of NAb appears to be directly correlated with the dose of therapeutic IFN administered, up to a certain dose threshold. If the dose is increased beyond this threshold, the levels of NAb decrease. The biological significance of anti IFN NAb is not yet known, nor has it been proven conclusively that they affect the clinical response to IFN beta therapy. The presence of NAb is therefore not an indication that treatment should be changed. Indeed, any treatment decision should be based only on the clinical response to therapy. PMID- 15353282 TI - Vascular elastic laminae: anti-inflammatory properties and potential applications to arterial reconstruction. AB - Biomaterials, including non-biodegradable and biodegradable polymers, and collagen and fibrin matrices, have been used in experimental and clinical arterial reconstruction. While these biomaterials exhibit various characteristics suitable for arterial reconstruction, the patency of biomaterial-based arterial substitutes remains problematic because of inflammation and thrombogenesis. Endothelial cell seeding of biomaterials has been proposed and used for reducing the thrombogenicity of biomaterials. However, difficulties in cell retention hamper the application of such an approach. Although autogenous vein grafts offer satisfactory results, not all patients possess veins available for arterial replacements. Thus, a critical issue in arterial reconstruction is developing arterial substitutes that are inflammation/thrombosis-resistant while possessing the characteristics of natural arteries. Here we show that allogenic vascular elastic laminae exhibit anti-inflammatory properties and may be considered a potential material for arterial reconstruction. In this article, we briefly review the composition, structure, and function of vascular elastic laminae, summarize recent discoveries on the role of elastic laminae in regulating leukocyte adhesion and vascular smooth muscle cell proliferation and migration, and discuss potential applications of allogenic elastic laminae to arterial reconstruction. PMID- 15353283 TI - Multiple functions of maspin in tumor progression and mouse development. AB - Maspin is a unique serpin with diverse biological functions. Initially identified from human normal mammary epithelial cells, maspin expression is either reduced or completely silenced in breast cancers. Numerous studies have implicated maspin function in cancer progression and angiogenesis. Maspin has also been targeted for breast cancer gene therapy. Recently, transgenic and gene knockout mouse models have been used by our laboratory to identify the biological functions of maspin in vivo. In this review, we summarize the multiple functions of maspin in tumor progression and mouse development. These data demonstrate that maspin not only plays a role in tumor progression and metastasis but also is a key regulatory molecule for normal mammary gland and embryonic development. PMID- 15353284 TI - Molecular mechanisms of NCAM function. AB - Neural cell adhesion molecule (NCAM) was originally characterised as a homophilic cell adhesion molecule (CAM) abundantly expressed in the nervous system. However, the last decade of research has challenged the traditional view and defined novel roles for NCAM. NCAM is now considered a signaling receptor that responds to both homophilic and heterophilic cues, as well as a mediator of cell-cell adhesion. This review describes NCAM function at the molecular level. We discuss recent models for extracellular ligand-interactions of NCAM, and the intracellular signaling cascade that follows to define cellular outcomes such as neurite outgrowth. PMID- 15353285 TI - Kaposi's sarcoma-associated herpesvirus immediate early gene activity. AB - KSHV is the causative agent of three human proliferative disorders: Kaposi s sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. Herpesvirus gene expression and viral replication is a complex, tightly regulated process involving latent, immediate early, early, and late viral gene transcription. The immediate early genes generally code for transcriptional activators and are critical for initiating viral transcription. KSHV encodes for approximately nine immediate early gene products, including ORF50, K8, K9, K3, K5, ORF57, ORF29b, ORF45, and K4.2. This review will address the activities of these proteins and what roles they play in virus replication, evasion of the host immune response, and viral pathogenesis. PMID- 15353286 TI - The role of TWEAK/Fn14 in the pathogenesis of inflammation and systemic autoimmunity. AB - Interactions between members of the TNF ligand superfamily with their cognate TNF receptors play a crucial role in maintaining immune homeostasis in normal individuals, while dysregulation of certain TNF-ligands and receptors contributes to the pathogenesis of autoimmunity. Identification of novel members of the TNF ligand and receptor families will promote our understanding of the pathogenesis of systemic autoimmune diseases, thus facilitating the development of novel therapeutic approaches. TNF-like weak inducer of apoptosis (TWEAK), a recently identified member of the TNF ligand family, induces PGE2, MMP-1, IL-6, IL-8, RANTES, and IP-10 in fibroblasts and synoviocytes, and upregulates ICAM-1, E selectin, IL-8, and MCP-1 in endothelial cells. The receptor for TWEAK, Fn14, is expressed in various organs including the kidney; it is intriguing that some of these chemokines induced by TWEAK are crucial in the pathogenesis of lupus nephritis. Furthermore, others have described upregulated TWEAK expression on the surface of T cells in human lupus. In this paper we review the possible roles of TWEAK/TWEAK receptor interactions in the pathogenesis of inflammatory and systemic autoimmune diseases, with particular focus on systemic lupus erythematosus. TWEAK blockade may be helpful therapeutically in restoration of tolerance, but is more likely to modify inflammatory damage in target organs. PMID- 15353287 TI - Crystal structures of cyclophilin and its partners. AB - Cyclophilin (CyP) is a cytosolic receptor of immunosuppressive drug cyclosporin A (CsA). The binary complex of CyP-CsA inhibits the activity of Ca2+/calmodulin dependent serine/threonine calcineurin (CN). The inhibition of CN in turn disables the transcription activity of nuclear factor of activated T cell, thus suppressing the T cell activation and cardiac hypertrophy. CyP is also an enzyme catalyzing peptidyl-prolyl cis-trans isomerization and serves as a molecular chaperone in various biological processes. For example, CyPA is involved in the assembly/deassembly of HIV-1 virion and is required for the full infectious activity of HIV-1. However, the in vivo function of CyP remains a mystery. This review will describe the three-dimensional structures of CyPs and its partners and discuss the structural clues to understanding the CyP functions in biological processes. The structures of CyP in complex with proline-containing peptides provided insight into the mechanism of peptidyl-prolyl cis-trans isomerization. The structures of CyPA in complex with HIV-1 capsid protein and its peptides revealed details of interactions of CyP with HIV-1 capsid protein, thus providing a guideline for design of anti-HIV drugs. The rearrangement of two tetratricopeptide repeats of the, large, cyclophilin CyP40 into a long helix under the crystallization conditions might be biologically relevant to the CyP40 function in the hsp90 molecular chaperone system. The structures of the binary CyPA-CsA and ternary CN-CyPA-CsA complexes showed how CsA binds to its receptors and therefore provide a template for design of new immunosuppressive drugs. PMID- 15353288 TI - The febrile patient: diagnostic, prognostic and therapeutic considerations. AB - Although clinicians have long pondered the diagnostic and prognostic implications and the treatment of fever, fundamental questions remain unanswered. The value of the height or pattern of a fever in predicting the etiology or course of the illness causing it is a case in point. Whether fever is ever harmful and should, therefore, be suppressed is another. These controversies and others concerning the febrile patient are the subject of this manuscript. PMID- 15353289 TI - Novel approaches and cutting edge immunotherapies in multiple sclerosis. AB - MS is a chronic inflammatory disease of the central nervous system (CNS). MS is a predominantly CD4+ T cell mediated autoimmune disorder. Recent studies have challenged this existing paradigm by supporting the role of other immune cells and factors (even non-immune) including CNS antigen-driven clonally expanded B cells, autoantibodies, complement and mediators of the innate immune responses in MS lesions. Further expansion of this global CNS dysfunction includes oligodendroglial cell (OGC) loss, attenuated remyelination, axonopathy, and gliosis. The recognition of new "players" directing effector and regulatory functions and further insight into reparative mechanisms occurring at various stages of the disease within a given individual will influence ongoing and future therapeutic trials. The following discussion will encompass evolving concepts in the pathogenesis of MS with a focus on novel immunotherapies. These new approaches reflect targeting of a multifaceted spectrum of immune activity. The immunotherapies will be characterized by their intervening role of specific and/or multiple pathogenic steps including initiation, peripheral activation, molecular co-stimulation and immune effector responses during early, transitional and late phases of disease. Emerging strategies for the enhancement of neuroprotection and reparative mechanisms will also be reviewed. Classification of novel approaches will include the following main types of immunotherapies: (1) targeting of myelin specific T cells: antigen-specific therapies (2) targeting of B cell and autoantibody responses (3) targeting of immunologic steps of disease pathology (4) targeting of reparative stages of disease: neurotrophic and neuroprotective, (5) global therapies: broad-based polydirectional strategies. PMID- 15353290 TI - Synovial activation in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease with progressive articular damage. Activated cells of the synovium produce pro-inflammatory and matrix-degrading effector molecules, which maintain the inflammation and lead to the destruction of the involved joints. In addition to macrophages and T- and B cells, fibroblast-like synoviocytes must be considered key cells in driving the pathological processes. They can be distinguished by their transformed-appearing phenotype and their invasion into adjacent cartilage and bone. Synovial activation is driven by pro-inflammatory cytokines as well as cytokine independent pathways including endogenous retroviral elements and Toll-like receptors (TLR). These pathways are connected by a complex network of autocrine and paracrine acting factors. Another feature of RA synovium is hyperplasia of the lining layer, which results from increased proliferation and decreased apoptosis of synovial fibroblasts. Thanks to new techniques in basic research, novel insights into the cellular and molecular mechanisms of the pathogenesis of RA were gained and led to the development of new, specific therapeutic strategies. PMID- 15353291 TI - Enterococcal virulence--pathogenicity island of E. Faecalis. AB - Enterococcal species, though most commonly regarded as members of the microbial flora of the intestinal tract, have recently emerged as human pathogens of significant concern. The rapid spread of antibiotic resistance among enterococci, which has resulted in strains now being routinely isolated that are resistant to all bactericidal regimens, has prompted considerable interest in investigating the pathogenesis of enterococcal infection, with a view toward deriving new, information-based treatment strategies. This review summarizes major findings on the pathogenesis of enterococcal infection, fits them into a model for the dual lifestyle of enterococci as commensal and pathogen, and integrates into that model a recently discovered pathogenicity island of Enterococcus faecalis. PMID- 15353292 TI - Mechanisms of HTLV-1 transformation. AB - HTLV-1 is the etiological agent of the fatal disease adult T-cell leukemia. The virus encodes many proteins including several accessory proteins, p12I, p13II, p27I, and p30II, whose roles have recently begun to be elucidated. These accessory proteins are important in T-cell activation, transcriptional regulation, viral persistence, and virus assembly. The viral oncogene Tax is thought to be largely responsible for tumorigenesis, although the precise mechanisms underlying transformation are not completely understood. Tax has a profound impact on transcription, cell growth regulation, genomic stability and apoptosis. This review will provide possible contributions of the accessory proteins to transformation as well as highlight the alterations of the above mentioned cellular events by Tax. Animal models of both Tax and the accessory proteins are also included based on the essential information on the transformation process in vivo that they provide. PMID- 15353293 TI - Current advances in molecular immunology: reference guide for reviews on molecular vaccines. AB - The field of immunology has made significant and rapid advances in the past two decades. This understanding has led to a systematic approach to studying the various aspects of development, activation, expansion, maintenance and termination of immune responses, with respect to disease-pathogenesis, prevention and therapeutic interventions. This issue of the journal focuses on reviewing the molecular aspects of vaccine design. In order to provide the reader with a framework and reference to the new aspects of immunological concepts, this article reviews the recent developments in immunology, with emphasis on induction of immune responses during vaccination. Key recent reviews in each section are provided for the readers to obtain detailed information. Several schematic figures are provided for a visual representation of basic concepts. PMID- 15353294 TI - Role of viral regulatory and accessory proteins in HIV-1 replication. AB - Human immunodeficiency virus-1 (HIV-1) is the causative agent of acquired immune deficiency syndrome (AIDS), a disease characterized by CD4+ T lymphocyte depletion. HIV-1 replicates actively in a variety of cells by encoding several regulatory (Tat and Rev) and accessory (Vpr, Vif, Vpu, and Nef) proteins. Accessory proteins, thought initially to be dispensable for infection, have now been shown to be important for efficient infection in vivo. Recent evidence suggests that certain viral proteins, like Vif, have evolved to overcome the antiviral mechanisms of the host, while proteins like Nef, which are markers for disease pathogenesis in vivo, help to increase pathogenesis by targeting bystander cells. Thus, these proteins control many aspects of the virus life cycle as well as host cell function, namely gene regulation and apoptosis. Understanding the mechanisms by which the virus is able to successfully replicate in host cells and subsequently cause gradual destruction of the immune system may yield new approaches for therapeutic strategies. In this review, we attempt to integrate information on the role of these regulatory and accessory proteins, emphasizing their interactions with other viral and cellular components, and the subsequent effect on viral replication. PMID- 15353295 TI - Functional role of acetylcholine in the immune system. AB - Increasing evidence indicates that the central nervous system (CNS) can operate certain immune functions. There is a bidirectional regulation system between the brain and peripheral immune system during infection and inflammation. Furthermore, autopsy brain preparations in Alzheimer's disease patients show the up-regulated expression of proinflammatory cytokines, suggesting an important role of cytokines in disease progression. Recently, acetylcholine has been suggested to inhibit lipopolysaccharide (LPS)- induced inflammation in macrophages. In this paper, we review the important aspects of several cytokines under the pathologic conditions of CNS. Furthermore, we discuss recent views on the nonneuronal action of acetylcholine in the cross talk between acetylcholine and the immune system. PMID- 15353296 TI - Prolyl isomerases in yeast. AB - Prolyl isomerases are enzymes that catalyze cis-trans isomerization of peptidyl prolyl bonds and span three structurally unrelated protein families: the cyclophilins, FKBPs, and parvulins. The genome of the budding yeast Saccharomyces cerevisiae encodes eight different cyclophilins (Cpr1 to Cpr8), four FKBPs (Fpr1 to Fpr4), and a single parvulin (Ess1). Remarkably, two of these proteins, cyclophilin A and FKBP12, are conserved from yeast to humans and mediate virtually all of the intracellular actions of the immunosuppressive antifungal drugs cyclosporin A, FK506, and rapamycin. The study of prolyl isomerases in S. cerevisiae has proven invaluable to understand the elusive functions of these proteins, and continues to provide new insights into their diverse cellular roles. Here we review the current state of knowledge about prolyl-isomerases in this model organism. PMID- 15353297 TI - Sickness behavior in fever and hypothermia. AB - Sickness behavior has become a common expression in the description of general symptoms of diseases and regarded as partly or fully advantageous for the patient to combat infection or other disturbance acting on the body. Several components of sickness behavior such as anorexia, sleepiness and inactivity have significant energetic connotations and hence may affect body mass and/or body temperature. Thermoregulatory accompaniments of sickness behavior could be either fever or hypothermia depending on the nature and severity of disease. A survey of the relevant literature has identified afferent, central and efferent mechanisms that may allow separate or coordinated appearance of behavioral and/or thermoregulatory aspects of these symptoms occurring under different experimental conditions. An attempt has been made to find some biological logic in the appearance of various components of sickness behavior and changes in body temperature that could explain the purported positive value of sickness behavior in disease survival. PMID- 15353298 TI - Passive immunization for immunocontraception: lessons learned from infectious diseases. AB - Development of vaccine for contraception is an exciting proposition that could provide a valuable alternative to the presently available methods for birth control. Various targets such as gonadotropin releasing hormone (GnRH), follicle stimulating hormone (FSH), leutinizing hormone (LH), zona pellucida (ZP) antigens, sperm antigens, and human chorionic gonadotropin (hCG) are being explored for immunocontraception. Besides specific concerns associated with each contraceptive vaccine, the progress has been restricted by the variability of the immune response after active immunization, attain and maintain high antibody titers, time lag to achieve reasonably good antibody titers, and uncertainty regarding how long the bioeffective antibodies will remain in circulation. It is envisaged that these concerns may be taken care of by using the preformed antibodies in the passive immunization approach. The antibody therapies have been tried and found to be successful against various infectious diseases both in animals as well as humans. Some have become treatment modalities in the clinics. This manuscript will review the data available for the passive immunization of preformed polyclonal and murine/humanized/human monoclonal antibodies, their efficacy, mode of delivery, duration of the effects, and limitations, if any. The overall objective is to examine the feasibility and practicability of the passive immunization approach for immunocontraception. PMID- 15353299 TI - Thymidylate synthase: a critical target in cancer therapy? AB - For the last four decades, synthesis and testing of potentially active drugs (e.g., antimetabolites) have focused on structural modification of existing metabolites as precursors of DNA and RNA synthesis. In recent years, the focus has shifted to synthesis of target-specific agents. Thus, the current emphasis of drug development is directed at inhibiting specific target(s) expressed preferentially, if not exclusively, in tumor tissues, with the ultimate goal of improving the therapeutic efficacy and selectivity of these new agents. Preclinically, proof-of-principle studies were carried out in tumors with specific expression of the intended target. With the hope of translating preclinical findings to the design of implementation of clinical trials. Thymidylate synthase (TS) continues to be a critical target for 5-fluorouracil (5 FU) and its prodrugs, UFT/LV (Orzel), capecitabine (Xeloda), and S-1, primarily because this enzyme is essential for the synthesis of 2-deoxythymidine-5 monophosphate, a precursor for DNA synthesis. While fluoropyrimidine antimetabolites have other sites of action, antifolates ZD1694 (raltitrexed, Tomudex) and AG337 (Thymitag) are more specific and potent TS inhibitors. Thus, it is hoped that pronounced and sustained inhibition of this enzyme could result in downstream regulation of molecular markers associated with sensitivity and resistance to these agents. It is also critical to recognize that the degree and duration of inhibition of the target enzyme may depend on the expression level of the target enzyme, thymidylate synthase. Correlative studies in preclinical and clinical systems demonstrated a close relationship between the enzyme level (mRNA and protein) and response to therapy of colorectal cancer patients treated with fluoropyrimidine or Tomudex. However, significant overlap was demonstrated between responders and non-responders. These data are consistent with the hypothesis that prediction of response to anticancer drugs is multifactorial, and TS is one target. Clinically, although overall response of colorectal cancer patients to a variety of TS inhibitors is similar, toxicity profiles are different. The availability of the 5-FU prodrugs offers the possibility of greater therapeutic selectivity based on the demonstration that thymidine phosphorylase, the activating enzyme for 5-FU, is expressed at a higher level in tumor tissue compared with normal tissue counterparts. It is likely that successful application of TS inhibitors will not only be based on measurement of the TS level in tumors vs. normal tissues, but on the delineation of the consequences of this inhibition on molecular markers associated with cellular proliferation, apoptosis and cell cycle regulation. PMID- 15353300 TI - MMPS and TIMPS in ovarian physiology and pathophysiology. AB - The matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) have been postulated to play a critical role in the extracellular matrix (ECM) remodeling associated with follicular development. The gelatinases were localized to the theca of developing follicles and in the stroma of the rodent ovary. Gelatinolytic activity corresponded with the localization of MMP-2 and MMP-9 around the developing follicles and at the apex of preovulatory follicles. The TIMPs-1, -2, and -3 were localized to the stroma and theca of developing follicles and correlation between MMPs and the quality of the developing follicles was found. During the process of ovulation, MMP-1 protein was found in the theca interna and externa, interstitial glands, and germinal epithelium. Synthetic inhibitor of mammalian tissue collagenases was documented to be inhibitory to ovulation in perfused rat ovaries. MMP-19 and TIMP-1 messenger RNA were localized to the granulosa and thecal-interstitial cells of large preovulatory and ovulating follicles. Both were induced and upregulated 5-10 fold by human chorionic gonadotropin (hCG). MMP-2 mRNA found in theca-interstitial cells and membrane-type (MT) 1-MMP mRNA found in granulosa and theca-interstitial cells were both induced after stimulation with pregnant mare's serum gonadotropin (PMSG). Gelatinolytic activity was observed throughout the formation of the corpus luteum. At 12 h after hCG, luteinizing granulosa cells expressed TIMP-1 and TIMP-3 mRNA. In the newly forming corpus luteum at 24 h after hCG administration, the luteal cells expressed TIMP-1, -2, and -3 mRNA with unique pattern of cellular expression for each of the TIMPs. Regression of the corpus luteum is associated with a significant increase in the activity of the metalloproteinases. In luteinized granulosa cells from women with polycystic ovarian syndrome (PCOS) the MMP-TIMP balance is shifted towards greater MMP activity. Cultured luteinized granulosa cells obtained from PCOS patients secrete higher levels of MMP-9 and MMP-2 compared to granulosa cells from normal ovulatory patients whereas the secreted basal level of TIMP-1 was similar in both types of granulosa cells. These results indicate a higher net gelatinolytic activity within the luteinizing granulosa cells of patients with PCOS. It has been shown that in sheep, diversion of normal follicles to atresia by hypophysectomy is followed by a significant increase of intrafollicular levels of MMP-2 and MMP-9 and the disappearance of connexin-43. It is therefore reasonable to speculate that MMP-9 and MMP-2 may be associated with inappropriate atresia in PCOS. PMID- 15353301 TI - Pharmacogenomics of thymidylate synthase in cancer treatment. AB - Cancer drugs such as 5-fluorouracil (5-FU) that target the enzyme thymidylate synthase (TS) have been and are still being widely used in cancer treatment, but as with other anti-cancer drugs, the majority of tumors do not respond to the treatment, whereas the patients still suffer drug-related toxicity. The most recent attempts at improving cancer treatment have taken the pharmacogenetic approach of identifying biochemical response determinants for response, so that patients with suboptimal determinants who unlikely to respond can be identified prior to treatment. Studies to date indicate that high intratumoral levels of TS gene expression or TS protein generally predict for non-response, whereas low levels are associated with a high response rate. Measuring these determinants requires tumor tissue and, in the case of gene expression, a technically demanding quantitative PCR procedure. Thus, considerable interest was generated by data suggesting that the variable number of a 28 base-pair (bp) segment in the promoter region of the TS gene was associated with TS gene expression and/or protein expression, as well as with tumor response to 5-FU therapy, toxicity and patient survival. However, not all studies have obtained the same results, so that the role of this TS polymorphism as a predictor of treatment outcome is still not clear and is currently under evaluation. This review will summarize pharmacogenomic studies of TS that were aimed at elucidating the function of this genetic polymorphism. PMID- 15353302 TI - Infectious transmission and replication of human T-cell leukemia virus type 1. AB - Retrovirus infection proceeds by attachment of the envelope glycoprotein to a cell surface receptor, followed by fusion of the viral and cellular membranes. Once in the cell, the viral enzymes and structural proteins form a replication complex that converts the single-stranded viral genomic RNA into a double stranded DNA, which is then integrated into the host cell chromosome. For HTLV-1, these events are not well characterized. We have developed cell culture systems, infectious molecular clones, and viral vectors that can be used to characterize the mechanisms of HTLV-1 infection and replication. Infection with cell-free HTLV 1 virions is orders of magnitude less efficient compared with other retroviruses. This inefficiency is the result of a block in the replication process after the virion is bound to the cell surface. We are determining whether this block is conferred by the viral replication enzymes, results from the actions of cellular restriction factors, reflects the need for cell-cell contact, or is caused by a combination of these factors. PMID- 15353303 TI - Navigating novel mechanisms of cellular plasticity with the NAD+ precursor and nutrient nicotinamide. AB - Interest in neuroprotectants for the central nervous system continues to garner significant attention. Nicotinamide, the amide form of niacin (vitamin B3), is the precursor for the coenzyme beta-nicotinamide adenine dinucleotide (NAD+) and is considered to be necessary for cellular function and metabolism. However, recent work has focused on the development of nicotinamide as a novel agent that is critical for modulating cellular plasticity, longevity, and inflammatory microglial function. The ability of nicotinamide to preserve both neuronal and vascular cell populations in the brain during injury is intriguing, but further knowledge of the specific cellular mechanisms that determine protection by this agent is required. The capacity of nicotinamide to govern not only intrinsic cellular integrity, but also extrinsic cellular inflammation rests with the modulation of a host of cellular targets that involve protein kinase B, glycogen synthase kinase-3 beta (GSK-3 beta), Forkhead transcription factors, mitochondrial dysfunction, poly(ADP-ribose) polymerase, cysteine proteases, and microglial activation. Intimately tied to the cytoprotection of nicotinamide is the modulation of an early and late phase of apoptotic injury that is triggered by the loss of membrane asymmetry. Identifying robust cytoprotective agents as nicotinamide in conjunction with the elucidation of the cellular mechanisms responsible for cell survival will continue to solidify the development of therapeutic strategies against neurodegenerative diseases PMID- 15353304 TI - Translational autoregulation of thymidylate synthase and dihydrofolate reductase. AB - The folate-dependent enzymes, thymidylate synthase (TS) and dihydrofolate reductase (DHFR) are critical for providing the requisite nucleotide precursors for maintaining DNA synthesis and DNA repair. In addition to their essential roles in enzyme catalysis, these two enzymes have now been shown to function as RNA binding proteins. Using in vitro and in vivo experimental model systems, we have shown that the functional consequence of binding of TS protein to its own cognate mRNA, as well as binding of DHFR to its own DHFR mRNA, is translational repression. Herein, we review and update studies focusing on the translational autoregulatory control of TS and DHFR expression and discuss the molecular elements that are required for these specific RNA-protein interactions. Moreover, we present evidence showing that abrogation of these normal translational autoregulatory feedback mechanisms provides the molecular basis for the rapid development of cellular drug resistance. PMID- 15353305 TI - Immunopathogensis of HTLV-I associated neurologic disease: molecular, histopathologic, and immunologic approaches. AB - Human T-cell lymphotropic virus type I (HTLV-I) infection is associated with a variety of human diseases including HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP), a chronic progressive inflammatory neurological disease. An important risk factor for the development of HAM/TSP is thought to be a high HTLV-I proviral load. Histopathological studies have demonstrated the presence of HTLV-I virus in the affected areas of spinal cords from HAM/TSP patients. Furthermore, T-cell infiltrations have been shown in spinal cord lesions. The precise mechanism for disease development is still unknown. Virus-host immune interactions are considered to play an important role in disease pathogenesis. This review focuses on current molecular, histopathological, and immunological approaches to understand the immunopathogenesis of HAM/TSP. PMID- 15353306 TI - Brain RNA expression in obese vs lean mice after LPS-induced systemic inflammation. AB - Mortality of obese patients with severe sepsis is higher than non-obese patients. Thus far, a pathophysiologic mechanism has not been identified that explains this higher mortality. The central nervous system is now becoming increasingly recognized as a target organ in sepsis and the systemic inflammatory response syndrome and may hold clues to the deleterious affects of obesity in patients with sepsis syndrome. In this study, obese and non-obese mice were given LPS IP and the brains were harvested 2 hours after injection. The brains were processed and mRNA isolated and hybridized to a microarray chip and processed. Analysis of gene expression demonstrated distinct expression difference between the lean and obese animals. Ontology data supports clear differences between the lean and obese groups in the coagulation system, neuro-endocrine system, lipid transport and insulin receptors. Approximately eighty genes were identified to show 10-fold differential expression between the obese and lean mice. PMID- 15353307 TI - Prevention of colorectal cancer using COX-2 inhibitors: basic science and clinical applications. AB - Cyclooxygenase-2 (COX-2), an inducible prostaglandin G/H synthase, is overexpressed in pre-neoplastic tissues and several human cancers including colorectal cancer. Evidence linking COX-2 activity to carcinogenesis was derived from epidemiologic studies and animal models with defect adenomatous polyposis coli (APC) gene. PGE2 induced by COX-2 exerts several biological properties that may be advantageous for tumorigenesis: 1) Promoting angiogenesis (increased VEGF, bFGF, and PDGF production), 2) Anti-apoptosis mechanism (via increased bcl-2 and Akt activity), 3) Stimulating tumor metastasis (by increasing matrix metalloproteinases) and 4) Decreased immune surveillance (decreased cytokine production and NK activity). In addition, COX-2 reaction can cause DNA oxidation and induce mutations. Chemoprevention of colorectal cancer has attracted great attention in recent years. Epidemiologic data showed that chronic intake of traditional nonsteroidal anti-inflammatory drugs (NSAIDs) could reduce the incidence of colorectal cancer. Recent clinical trial studies showed that celecoxib, a selective COX-2 inhibitor, is equally effective in reducing colorectal adenomas in animal models and patients with familial adenomatous polyposis (FAP), yet with superior GI safety. Two COX-2 inhibitors (celecoxib and refocoxib) have been approved by FDA as adjuncts to usual care in FPA patients, and are currently being studied in patients with sporadic adenomas and other types of cancers. These studies are expected to generate evidence in favor of targeting COX-2 and its gene products as chemopreventive strategies, which may provide an alternative in current approach to reducing the morbidity and mortality of this disease. PMID- 15353308 TI - The role of isoflavones in cancer chemoprevention. AB - Cancer is one of the major health problems around the world. However, it has been estimated that more than two-thirds of human cancers could be prevented by modification of lifestyle including dietary modification.The incidences of hormone-related cancers are much higher in Western countries compared to Asian countries. One of the major differences in diet between these populations is that the Asians consume a traditional diet high in isoflavones. Epidemiologic evidence together with data from animal and in vitro studies strongly supports relationship between isoflavones and the lower risk of cancers. Isoflavones have been shown to inhibit carcinogenesis in vivo in animal experiments. It has been known that genistein, one of the major isoflavones, inhibits the growth of various cancer cells through the modulation of genes that are related to the control of cell cycle, apoptosis, and cell signaling pathways. Moreover, genistein has been found to be a potent inhibitor of oxidative stress, angiogenesis, and metastasis.Therefore, isoflavones exert beneficial effects on human health and may be promising agents for cancer prevention and/or treatment. However, further in depth experimental investigations along with clinical trials are needed to fully evaluate the value of isoflavones in human cancer prevention and/or treatment. PMID- 15353309 TI - Chemopreventive role of folic acid in colorectal cancer. AB - Mortality from colorectal cancer, a leading cause of death in the U.S.A. and other western countries, has remained unchanged over the past 45 years. Therefore, the search for strategies to prevent the development and progression of colorectal cancer has markedly intensified. Chemoprevention is one such strategy. Accumulating evidence suggests that folic acid, a water soluble vitamin, could be an effective chemopreventive agent for colorectal cancer. Results from several studies have demonstrated that a diet deficient in folic acid may be associated with an increased risk of colonic neoplasia, whereas dietary supplementation of this nutrient may be chemopreventive. Although the mechanisms by which folic acid exerts its chemopreventive role in colorectal carcinogenesis remain to be fully elucidated, supplemental folic acid has been shown to arrest the loss of heterozygosity (LOH) of the tumor suppressor gene DCC (deleted in colorectal cancer) and to stabilize its protein in normal appearing rectal mucosa of patients with colorectal adenomas. Data from in vitro studies utilizing colon cancer cell lines suggest that supplemental folic acid or its metabolite 5-methyltetrahydrofolate (5-MTF) attenuates the expression and activation of EGF-receptor (EGFR) as well as proliferation of cells. The folic acid mediated reduction of EGFR function could partly be the result of suppression of EGFR gene through increased methylation of CpG sequences within its promoter. PMID- 15353310 TI - The promise and challenge toward the clinical application of maspin in cancer. AB - Since the identification of the human maspin gene, a decade of extensive research revealed the promise of maspin both as a valuable molecular marker for the diagnosis and prognosis of many types of cancers, and as a tumor suppressor at the level of tumor growth, invasion, angiogenesis, metastasis, and tumor sensitivity to drug-induced apoptosis. This review is intended to summarize the consensus of these findings, and provide an overview of the current challenges toward the clinical application of maspin. Specifically, this review discusses several likely molecular mechanisms underlying (1) the differential regulation of maspin expression, and (2) the biological activities of maspin in tumor progression. PMID- 15353311 TI - Orexigenic vs. anorexigenic peptides and feeding status in the modulation of fever and hypothermia. AB - Prevailing changes in the feeding status or the nutritional status, in general, can modify the expression of many orexigenic and anorexigenic peptides, which influence hypothalamic functions. These peptides usually adjust body temperature according to anabolic (increased appetite with suppressed metabolic rate and body temperature) or catabolic (anorexia with enhanced metabolism and temperature) patterns. It was plausible to presume that such peptides contribute to regulated changes of body temperature (either fever or hypothermia) in systemic inflammation, particularly since anorexia is a common feature in inflammatory processes. No consistent, common, or uniform way of action was, however, demonstrated, which could have described the effects of various peptides. With the exception of cholecystokinin (CCK), all investigated peptides were devoid of real thermoregulatory actions: they influenced the metabolic rate (and consequently body temperature), but not the mechanisms of heat loss. Central CCK is indeed catabolic and may participate in febrigenesis. Leptin may activate various cytokines, catabolic peptides and may inhibit anabolic peptides, but it probably has no direct febrigenic effect and it is not indispensable in fever. Melanocortins and corticotropin-releasing factor provide catabolic adaptive mechanisms to food intake (diet induced thermogenesis) and environmental stress, respectively, but they act rather as endogenous antipyretic substances during systemic inflammation, possibly contributing to the mechanisms of limitation of fever. Bacterial lipopolysaccharides enhance the expression of most of these catabolic peptides. In contrast, neuropeptide Y (NPY) expression may not be changed, only its release is decreased at specific nuclei, a defective NPY effect may also contribute to the febrile rise in body temperature. The data provide no clear-cut explanation for the mechanism of hypothermia seen in systemic inflammation. According to speculations, a presumed, overflow,-type release of NPY from the hypothalamic nuclei, as well as a suppression of the activity of catabolic peptides, could possibly cause hypothermia. There are no cues, however, referring to the identity of factors that could trigger such changes during systemic inflammation in order to induce hypothermia. PMID- 15353312 TI - Atherosclerosis as an inflammatory disease: implications for therapy. AB - Cardiovascular disease remains a leading cause of mortality in the United States despite the use of new pharmacologic therapy, lifestyle modifications, and different coronary interventions. Atherosclerosis represents a wide variety of pathologic lesions with different clinical impacts. In this review, we address the current understanding of the pathophysiological mechanisms underlying the development of atherosclerosis. We define atherosclerosis as a multifactorial process representing a series of molecular and cellular mechanisms and involving multiple interactions between lipid metabolism, monocyte activation, endothelial cells, cytokines and/or other intracellular metabolic pathways. We also imply that control of atherosclerosis could be achieved through therapeutic interventions at different sites of the inflammatory process. Therapeutic targets could include cytokine pathways, growth factors, transcription factors, defective genes and other intracellular metabolic pathways. PMID- 15353313 TI - Computational protein chemistry of p53 and p53 peptides. AB - Computational protein chemistry has potential to contribute to the development of new therapeutic approaches in medicine in several different ways, including indirectly by increasing understanding of the disease-associated changes in protein structure that are mechanistically important, which can have diagnostic implications, as well as directly in designing peptides to counteract the patho physiologic effects of these changes. Studies of the role of the tumor suppressor protein p53 in the carcinogenic process provide examples of both types of contribution. Computational studies of the effects of mutations in p53 on its structure have provided insights into cancer mechanisms and have served to elucidate potential new diagnostic approaches based on the identification of changes in p53 structure. Computational studies of p53 peptides have contributed to identifying and optimizing the structural characteristics that contribute to their activity in selectively killing cancer cells. PMID- 15353314 TI - Structure-based development of a novel collagen inhibitor for MMP-1: re-designing the functions of a matrix protein. AB - Collagenases are a highly specific class of enzymes. In their native states, collagenases cleave only native triple helical collagen molecules at a single peptide bond between Gly775-Leu776 for Type I collagen and Gly775-Ile776 for Type II collagen. The linear sequence of collagen is about 1050 amino acids in length, where three linear peptide sequences are required to form a triple helical collagen molecule. At present, there exist no crystallographic structures of collagenase bound to native triple helical collagen; nor has it been shown that collagenase recognizes the triple helical conformation of collagen. In our study, we have used an inhibitor design structure-activity based approach to show that collagenase recognizes and cleaves triple helical collagen conformations in preference to non-triple helical collagen conformations. PMID- 15353315 TI - Molecular modeling of mammalian cytochrome P450s. AB - The cytochrome P450 enzymes collectively metabolize a wide range of xenobiotic and endogenous compounds. The broad substrate specificity of this superfamily derives from the multiplicity of P450s whose unique substrate specificity profiles reflect underlying differences in primary sequence. Experimental structures of P450s, where available, have provided great insight into the basis of substrate recognition. However, for those mammalian P450s whose structures have not been determined, homology modeling has become an increasingly important tool for understanding substrate specificity and mechanism. P450 modeling is often a challenging task, owing to the rather low sequence identity between target and template proteins. Although mammalian P450 models have previously been based on bacterial P450 structures, the recent advent of mammalian P450 structures holds great potential for generating more accurate homology models. Consequently, the substrate recognition properties of several mammalian P450s have been rationalized using the predicted substrate binding site of recently developed models. This review summarizes the major concepts and current approaches of molecular modeling of P450s. PMID- 15353317 TI - Signaling the brain in inflammation: the role of endothelial cells. AB - Peripheral inflammation signals the brain primarily via blood-borne proinflammatory cytokines, released from activated immune cells. In addition to these cytokines, immune-brain signaling is known to involve another key mediator, prostaglandin E2 (PGE2), the level of which is elevated in the brain during various inflammatory states and which acts to influence the central neuronal activity to evoke some, but not all, of the sickness behavior including fever and the activation of hypothalamo-pituitary-adrenal axis. Studies over the last decade have indicated that brain endothelial cells are the major source of PGE2 under various inflammatory states. In this review, we highlight the significance of the endothelial mechanism in immune-brain signaling mediated by PGE2, but discuss also the possible influence of other mechanisms on brain PGE2 elevation. PMID- 15353316 TI - Conformational searches elucidate effects of stereochemistry on structures of deoxyadenosine covalently bound to tumorigenic metabolites of benzo[C] phenanthrene. AB - Remarkably different conformations can result when DNA binds with stereoisomeric compounds containing differing absolute configurations of substituents about chiral carbon atoms. Furthermore, the biochemical functions of covalent adducts with DNA are strongly affected by the stereochemistry of the ligands. Such stereochemical effects are manifested by DNA covalent adducts derived from metabolites of the non-planar fjord region environmental chemical carcinogen benzo[c]phenanthrene. To analyze these phenomena, an extensive conformational investigation for R and S stereoisomeric adducts to deoxyadenosine, derived from trans addition of enantiomeric anti diol epoxide metabolites of benzo[c]phenanthrene, has been carried out. We have surveyed the potential energy surface of the two adducts by varying systematically at 5 degree intervals in combination, the three important torsion angles that govern conformational flexibility of the carcinogen bulk with respect to the linked nucleoside. We carried out a grid search by creating 373, 248 structures for each isomer, and evaluated their molecular mechanical energies. This has permitted us to map the potential energy surface of each adduct in these three variables, and to delineate their low energy regions. The maps have a symmetric relationship which stems from the near mirror-image stereochemistry in the R and S isomers. This produces near mirror-image low energy structures in the nucleoside adducts. The limited sets of stereoisomer-dependent conformational domains delineated are determined by steric effects. Moreover, these features have been experimentally demonstrated to play governing structural roles in such carcinogen-damaged DNA duplexes: opposite orientations in the stereoisomer pairs computed for the nucleosides are observed by high-resolution NMR in the similarly modified DNA double helices, and are likely to play important roles in their interactions with enzymes involved in DNA transactions, and hence their biological activities. PMID- 15353318 TI - Tamoxifen: an emerging preventive. AB - Tamoxifen is well known for its actions as an antagonist of estrogen receptor mediated signaling and is one of the most extensively used endocrine agents both in the clinic and in the research setting. Tamoxifen has emerged from recent Breast Cancer Prevention Trials, conducted to evaluate risk reduction, as an effective preventive agent. Specifically, comparing tamoxifen to placebo (for 5 years) has shown that tamoxifen: (a) significantly reduced the risk of breast cancer recurrence, in those with a history of the disease; (b) reduced or delayed breast cancer progression, from an noninvasive to invasive breast cancer; (c) prevented or substantially reduced the risk of getting breast cancer (risk of occurrence) in healthy women with risk factors. The extraordinary outcomes offer support for the use of tamoxifen in multilevel preventive approaches and predict that it will continue to be vital in facilitating mechanistic studies. Information produced by mechanistic studies is needed to understand how to prevent cancer and how to confront treatment problems such as resistance. Molecular determinants of the resistant phenotype to tamoxifen are currently being identified. The next major effort will be to link these determinants to readily detectable biological changes that could be used to indicate the development of resistance before clinical manifestations develop. PMID- 15353319 TI - Identification of interstitial cells of Cajal in the human rectum. AB - It is postulated that the electric waves of the gut are generated by interstitial cells of Cajal (ICC). We hypothesized the presence of ICC in the rectum as initiating the rectal electric activity. The current study investigated this hypothesis. Two rectal specimens were taken from healthy areas of excised rectum of 22 rectal cancer patients (age 44.6+/-.2 SD years, 12 men, 10 women). The study specimens were subjected to c-kit immunohistochemistry. Controls for antisera specificity consisted of tissue incubation with normal rabbit serum substituted for the primary antiserum. C-kit positive branched ICC-like cells were detected in the rectal musculature of the studied specimens. They were distinguishable from the c-kit-negative non-branched smooth muscle cells and from the c-kit positive but non-branched mast cells. Immunoreactivity was absent in the negative controls. We have identified in the rectum for the first time cells with morphologic and immunologic phenotypes similar to the ICC of the gut. The role of these cells in normal physiologic and pathologic conditions of the rectum needs further studies. PMID- 15353321 TI - Activating the ryanodine receptor with dihydropyridine receptor II-III loop segments: size and charge do matter. AB - Excitation-contraction coupling in skeletal muscle is thought to depend on a physical interaction between II-III loop of the alpha1 subunit of the skeletal dihydropyridine receptor (DHPR) and ryanodine receptor (RyR). A peptide corresponding to II-III loop residues 671-690 of the skeletal DHPR (peptide A) is a high affinity activator of the RyR when it adopts an alpha-helical structure with critical basic residues aligned along one helical surface (1). Neither the structure of the full length II-III loop, or of sequences longer than 671-690 residues have been determined. Here we describe the structure and function of a 40 amino acid peptide corresponding to residues 671-710 (peptide AB) of the skeletal DHPR alpha1 subunit. This peptide contains the A region with a further 20 residues towards the C-terminus of the II-III loop. We predicted that peptide AB would strongly activate the RyR, because (a) it contains the active A sequence of basic residues and (b) it contains a greater proportion of the II-III loop. The structure of the AB peptide was determined and it was found to consist of two helical regions joined by an unstructured linker region. Surprisingly, although the structure of the A region was maintained, the 40 residue peptide was unable to release Ca2+ from skeletal SR. Strong activity was restored when four negatively charged residues in the C-terminal part of the peptide were replaced by neutral residues. The charge substitution caused minimal changes in the overall structural profile of the peptide and virtually no changes in the A portion of the peptide. The results suggest that the ability of the A region of the skeletal II-III loop to interact with the RyR could depend on the tertiary conformation of the II-III loop, which is thought to change during EC coupling. PMID- 15353320 TI - Adult T cell leukemia lymphoma. AB - Adult T cell leukemia lymphoma (ATLL) is a CD4+ lymphoproliferative malignancy resulting from human T-cell leukemia virus type 1 (HTLV-1) infection. It includes differing clinical forms classified as smoldering, chronic, lymphomatous, and acute ATLL. The Tax protein of HTLV-1 has been implicated as a viral oncoprotein which enhances virus replication and alters cellular gene expression, including activation of nuclear factor kappa B (NF kB), to result in lymphoid transformation. Chemotherapy for ATLL has had limited efficacy with median survivals of about 1 yr. Antiviral therapy employing zidovudine and interferon has shown promising results, as have antibody-based therapies to the interleukin 2 (IL2) receptor. Novel approaches employ a combination of chemo/antiretroviral therapy, hematopoietic stem cell transplantation, or inhibitors of NF kB activation. PMID- 15353323 TI - Obstructive sleep apnea, inflammation, and cardiopulmonary disease. AB - Obstructive sleep apnea (OSA) occurs commonly in the U.S. population and is seen in both obese as well as non-obese individuals. OSA is a disease characterized by periodic upper airway collapse during sleep, which then results in either apnea, hypopnea, or both. The disorder leads to a variety of medical complications. Neuropsychiatric complications include daytime somnolence, cognitive dysfunction, and depression. Increased incidence of motor vehicle accidents has been documented in these patients and probably reflects disordered reflex mechanisms or excessive somnolence. More importantly, vascular disorders such as hypertension, stroke, congestive cardiac failure, arrhythmias, and atherosclerosis occur frequently in these patients. The lungs may be affected by pulmonary hypertension and worsening of asthma. Recent data from several laboratories demonstrate that obstructive sleep apnea is characterized by an inflammatory response. Cytokines are elaborated during the hypoxemic episodes leading to inflammatory responses as marked clinically by elevated C-reactive protein (CRP). As elevated CRP levels are considered markers of the acute phase response and characterize progression of vascular injury in coronary artery disease, it is likely that obstructive sleep apnea could lead to worsening of vasculopathy. Moreover, as inflammatory mechanisms regulate bronchial asthma, it is also likely that cytokines and superoxide radicals generated during hypoxemic episodes could exacerbate reactive airway disease. Patients with Cough, Obstructive sleep apnea, Rhinosinusitis, and Esophageal reflux clustered together can be categorized by the acronym, "CORE", syndrome. The purpose of this manuscript is to review the inflammatory responses that occur in patients with obstructive sleep apnea and relate them to the occurrence of cardiopulmonary disease. PMID- 15353322 TI - Polymerisation underlies alpha1-antitrypsin deficiency, dementia and other serpinopathies. AB - We review here the molecular mechanisms that underlie alpha1-antitrypsin deficiency and show how an understanding of this mechanism has allowed us to explain the deficiency of other members of the serine proteinase inhibitor or serpin superfamily. These include the deficiency of antithrombin, C1-inhibitor and alpha1-antichymotrypsin in association with thrombosis, angio-oedema and emphysema respectively. Moreover the accumulation of mutant neuroserpin within neurones causes the novel dementia familial encephalopathy with neuroserpin inclusion bodies (FENIB). We have grouped these conditions together as the serpinopathies as recognition of their common pathophysiology provides a platform to develop strategies to treat the associated clinical syndromes. PMID- 15353324 TI - Major factors modulating the serum oxalic acid level in hemodialysis patients. AB - Ascorbic acid overload and vitamin B6 deficiency have been implicated in the development of hyperoxalemia in dialysis patients, but there is still disagreement about this. Hemodialysis patients who are exposed long-term hyperoxalemia may develop secondary oxalosis with an increased risk of cardiac, vascular, and bone disease, and thus may benefit from maintaining a low serum oxalic acid level. In 452 hemodialysis patients, the serum level of oxalic acid was 47.2 +/- 22.9 micromol /l before and 16.9 +/- 10.5 micromol/l after a 4-hour dialysis session, while the ascorbic acid levels were 39.0 +/- 92.7 micromol/l and 6.5 +/- 18.6 micromol/l, the glycolic acid levels were 7.3 +/- 10.1 micromol/l and 0.6 +/- 2.3 micromol/l, and the citric acid levels were 141.3 +/- 54.7 micromol/l and 117.6 +/- 37.2 micromol/l, respectively. Most patients (65.3 percent) had low serum ascorbic acid levels (less than 10 micromol/l) before hemodialysis. The serum level of oxalic acid [Ox] showed a significant positive correlation with the levels of ascorbic acid [AA], glycolic acid [Gly], and creatinine [Cre]: [Ox] = 21.711 + 0.181 x [AA] + 0.174 x [Gly] + 0.171 x [Cre], (all micromol/l, p less than 0.05). In 124 dialysis patients, the 4-pyridoxic acid level was 8.9 +/- 19.6 micromol /l before and 3.9 +/- 8.8 micromol/l after dialysis, and it was not correlated with oxalic acid or glycolic acid. Most dialysis patients (65.3 percent) had low serum levels of ascorbic acid, but a subgroup of patients (12 percent) had high serum ascorbic acid levels (more than 100 micromol/l) associated with hyperoxalemia (88.2 +/- 24.5 micromol/l). High dose vitamin C supplementation may aggravate hyperoxalemia in hemodialysis patients, so attention should be paid to avoiding this risk. PMID- 15353325 TI - Sensing and adapting to environmental stress: the archaeal tactic. AB - Archaea occupy a considerable diversity of niches encompassing extreme environments with extreme of pH, salinity and temperature that cannot be tolerated by other forms of life. Survival and colonisation requires the capacity to sense, and adapt to environmental change. In this review we consider the issues of adaptation to environmental stresses, in particular the mechanisms that might be employed by different Archaea to respond to the specific challenges of their particular niche. We lay emphasis on the strategies adopted to respond to oxidative and chemical stress. In particular, this paper reviews major key points in the generation of reactive oxygen species in Archaea, defense mechanisms and genetic responses to oxidative stress. Finally, we discuss complex biological response mechanisms to chemical damage with particular attention to detoxification from metals and drugs. PMID- 15353326 TI - Isoflavones in breast cancer chemoprevention: where do we go from here? AB - Based on the evidence from epidemiological, animal, in vitro data and human clinical trials, it is evident that isoflavones are promising agents for breast cancer chemoprevention. It is also evident that the form of isoflavone used (purified vs soy products), dose of isoflavone used (low vs high), timing and duration of exposure of isoflavones appears to play a major role in determining agonistic or antagonistic effects. Collectively, these isoflavones have enough evidence to warrant use in a number of clinical trials to examine its efficacy as a potential chemopreventive agent for breast cancer. In this comprehensive review, we attempt to summarize the evidence demonstrating the potential use of isoflavones in breast cancer chemoprevention and the rationale to examine a combination of biochemical, morphological and molecular intermediate endpoint biomarkers of breast cancer risk to examine the efficacy of this promising agent. PMID- 15353327 TI - Analytic approaches to differential gene expression in AIDS versus control brains. AB - We previously showed that specific strains of human immunodeficiency virus (HIV) 1 infect the brain and contribute to Neuropathology, Cognitive Distress, and Neuropsychiatric Disease. To study further brain disease that results from HIV-1 infection, we commenced analysis of changes in gene expression in brain. We analyzed RNA purified from Frontal Cortex of 5 HIV-1 infected and 4 HIV-1 negative control subjects RNA was amplified and Affymetrix technology was used to analyze gene expression using the 12,585 gene Affymetrix Human Genome U95A chip. The expressed genes showed highly significant Pearsons correlations with each other within the two groups. Expression intensities were transferred to Microsoft Excel and Spotfire was used to analyze the results. Twenty-group K-means cluster analysis was done for HIV+ and HIV- subjects. Genes that were expressed in the same cluster numbers in the two groups were removed from further analysis. Analysis of Gene expression in the top 13 HIV+ clusters showed expression in the 40 gene categories designated in our prior studies. Genes from several categories occurred in more than one K-means cluster. Genes identified in these lists included several genes that have been previously studied: MBP, Myelin-PLP, NMDA receptor, MAG, astrocytic protein, Notch 3, APP, Senescence, proteasome, Ferritin, signaling, cell cycle, iNOS, Chemokine, splicing, synapse, protein tags, and ribosomal proteins. The first (primary significant) axis of both Principal Component Analyses ordered the genes in the same patient groups as the K-means cluster analysis for the respective patient groups. PCA was thus not more informative than K-Means cluster analysis. Ratios of HIV+ to HIV- intensities were calculated for all the averaged gene expression intensities. The ratio range was 0.14 to 9.26. The genes at the extremes (ad extrema) did not correspond to the gene order by K-means clustering (or PCA). The genes in the top 13 K-means clusters showed low-level changes by expression ratio. Genes ad extrema by ratio were in clusters with very large memberships. Mann-Whitney analysis confirmed expression ratio results. Several inferences result from our preliminary study. First, study design will be different in future studies involving additional replicates. Second, ratios inform us of the extent of changes in gene expression quantitatively. Third, Cluster methodology provides us with more subtle information, how bunches (clusters) of genes behave in terms of their centroids (attractors). Fourth, genes that change extensively by ratio tend to be in the larger k-Means clusters. We conclude that ranking gene expression with the use of expression ratio or by K-means clustering, yield different representations of the data. PMID- 15353328 TI - Mechanisms and models of peripheral CD4 T cell self-tolerance. AB - Each response made by our immune system is either to promote or to prevent immune reactivity. That the immune system can successfully achieve both goals under specific biological conditions depends on central and peripheral tolerance mechanisms. Over the years, various experimental systems have been generated to study peripheral CD4 T cell tolerance. The experimental approaches are clearly diverse, but can be broadly categorized into those involving, tolerogenic, antigen injections, the use of transgenic mice (antigen transgenics, TCR transgenics and the combination thereof) and transplantation models. Results of these studies suggested a number of potential mechanisms mediating peripheral CD4 T cell tolerance, including clonal deletion, anergy and immune regulation. While the available systems all presented with their own limitations, they nevertheless provided a foundation from which our understanding of peripheral CD4 T cell tolerance will be built. In addition, previous studies done to examine CD8 T cell tolerance left with us some potentially helpful clues and ideas about how CD4 T cell tolerance should be studied. However, our current understanding on the immunity/tolerance decision made by CD4 T cells in the periphery remains incomplete. Evidence accumulated thus far favors the view that peripheral tolerance, unlike central tolerance, may not primarily determine the selection of specificities within the baseline T cell repertoire but may instead regulate cellular responses within the repertoire, in terms of both expansion/contraction and differentiation. Nevertheless, more robust and refined models need to be developed before we can make definitive conclusions about the potential role of peripheral tolerance in repertoire selection mediated by deletion of self reactive cells. PMID- 15353329 TI - Can ends justify the means? Digging deep for human fusion genes of prokaryotic origin. AB - Gene fusion has been described as an important evolutionary phenomenon. This report focuses on identifying, analyzing, and tabulating human fusion proteins of prokaryotic origin. These fusion proteins are found to mimic operons, simulate protein-protein interfaces in prokaryotes, exhibiting multiple functions and alternative splicing in humans. The accredited biological functions for each of these proteins is made available as a database at http://sege.ntu.edu.sg/wester/fusion/ PMID- 15353330 TI - Current vaccine adjuvants: an overview of a diverse class. AB - Numerous compounds are under evaluation as immunological adjuvants for improvement of vaccine performance. This review will briefly summarize some of the many diverse substances that are currently being utilized as vaccine adjuvants in preclinical and clinical studies. PMID- 15353331 TI - Epithelial cell invasion of the stroma in human skin organ culture. AB - Invasion of the surrounding tissue by malignant epithelial cells is a complex process. It involves degradation of basement membrane and stromal connective tissue elements and movement of tumor cells into and through the partially degraded matrix. While the process of tumor cell invasion has been extensively studied in simple two-dimensional or three-dimensional assay systems, it is difficult to know how the cellular and molecular mechanisms that underlie invasion in these culture systems relate to mechanistic events that drive invasion in intact tissue. To help address this question, our laboratory has developed a human skin organ culture model to study epithelial cell invasion of the stroma in intact tissue. In this model, punch biopsies of human skin are maintained in organ culture under serum-free, growth factor-free conditions or in the same culture medium supplemented with exogenous epidermal growth factor (EGF). In the absence of growth factor stimulation, normal tissue architecture and biochemical function are preserved. However, in the presence of exogenous EGF, the dermal-epidermal juncture is eroded and epithelial cells invade the dermis. Concomitant with dermal invasion is induction of matrix metalloproteinases (MMPs), stimulation of fibronectin synthesis and active epithelial cell migration into the stroma. These observations provide convincing evidence that events thought to be involved in invasion in simple two- and three dimensional in vitro models are also operative during invasion of intact tissue. PMID- 15353332 TI - Metal ion transport and regulation in Mycobacterium tuberculosis. AB - The regulation of metal ion concentrations is central to the physiology of the interaction between pathogenic bacteria and their hosts. Apart from the NRAMP orthologue, MntH, metal ion transporters in Mycobacterium tuberculosis have not been studied. Mn, the physiological substrate of MntH in other bacteria, may play an important role as a structural and redox-active cofactor in a wide range of metabolic processes. Fe, Cu and Zn play structural and catalytic roles in metalloenzymes involved in oxidative stress responses. Fe and Mg are required for growth in macrophages. Genomic analyses reveal 28 sequences encoding a broad repertoire of putative metal ion transporters (or transporter subunits), representing 24% of all transporters in this organism. These comprise 8 families of secondary active transporters and 3 families of primary active transporters, including 12,P, type ATPases. Potential metal ion specificities include K+, Na+, Cu2+, Cd2+, Zn2+, Mn2+, Mg2+, Ca2+, Co2+, Ni2+, Fe2+/3+, Hg2+, AsO2- and AsO4(2 ). 17 of these transporters are also encoded as complete open reading frames in Mycobacterium leprae, suggesting a role in intracellular survival. Iron transcriptionally regulates a diverse set of genes via the iron-dependent DNA binding proteins, Fur and IdeR. Changes in Fe and Mg concentrations signal entry into the intracellular compartment and potentially trigger up-regulation of virulence determinants. The plethora of putative transport systems encoded by the M. tuberculosis genome contrasts strikingly with the paucity of experimental data on these systems. The detailed analysis of the temporal pattern of M. tuberculosis transporter gene expression during infection will provide important insights into the basic biology of intracellular parasitism and may help to shape novel therapeutic strategies. PMID- 15353333 TI - Eight cytochrome P450s catalyze vitamin D metabolism. AB - Vitamin D3 plays a central role in calcium and phosphate homeostasis and is essential for the proper development and maintenance of bone. To exert its biological activities, vitamin D3 has to receive enzymatic transformation to the active form, 1,25-dihydroxyvitamin D3. The first step is the 25-hydroxylation reaction in the liver that produces 25-hydroxyvitamin D3, the major circulating form of vitamin D3. The 25-hydroxylation reaction is the prerequisite step for the subsequent 1 alpha-hydroxylation and 24-hydroxylation reactions in the kidney. The 1 alpha-hydroxylation reaction produces the active form of vitamin D3, whereas 24-hydroxylation reaction leads to inactivation. Both reactions are strictly controlled by parathyroid hormone, 1,25-dihydroxyvitamin D3, and calcium in a reciprocal manner in the kidney. At present, six cytochrome P450s (CYP2C11, 27A1, 2D25, 2R1, 3A4, and 2J3) are found to exhibit vitamin D 25-hydroxylation activities, and CYP27B1 and CYP24 are proved to be 1 alpha-hydroxylase and 24 hydroxylase, respectively. The main focus of this review is to summarize the properties of individual P450 in light of their catalytic activities to understand their physiological significance. PMID- 15353334 TI - B cell/antibody tolerance to our own antigens. AB - The lymphoid system normally mounts damaging responses to infectious pathogens while avoiding equally damaging responses to self. A notable number of antibodies to self antigens are formed but normally remain at levels below the damaging threshold, only temporarily rising to damaging levels during protective responses against infectious nonself. Many mechanisms regulate the level of autoantibodies and anti-self B cells including deletion, anergy, ignorance for antigen, receptor editing, coinhibition, competition for resources to sustain B cell responses, and apoptotic denouement of damaging responses following the ejection or containment of foreign invaders. While infectious events may encourage immune responses to self antigens, infectious events tend also to strengthen regulatory mechanisms. When regulatory mechanisms do not function properly, abnormal damaging responses to self antigens may occur. While defects in a single regulatory mechanism may result in autoimmunity, this eventuality usually happens only on permissive genetic backgrounds; this indicates that weakness in other regulatory mechanisms may be necessary to result in the emergence of damaging responses to self antigens. The immune system and its regulatory mechanisms are not simple, as one would expect of a homoeostatic process that also has the ability to expand enormously when challenged and to contract rapidly when threats pass. These processes that avoid damaging anti-self B cells are much more complicated than that envisaged in standard two signal models. Simple signals through the B cell antigen-receptor probably encourage B cell survival and receptivity, while other signals (costimulatory or coinhibitory) promote B cell stimulation or non stimulation/inactivation. PMID- 15353335 TI - Initiation of DNA replication in Xenopus egg extracts. AB - In the last decade, extraordinary advances in our understanding of the initiation step of eukaryotic DNA replication have been achieved. Many factors required for replication initiation have been identified, and an elegant model to explain how DNA replication is restricted to a single round per cell cycle has emerged. Of the many experimental approaches used to study DNA replication, egg extracts from Xenopus laevis are among the most powerful, since they recapitulate a complete round of cell-cycle regulated chromosomal DNA replication in vitro. In this review, we discuss current models for how DNA replication is initiated and regulated in Xenopus eggs, and we highlight similarities and differences seen between this and the other most common experimental organisms, yeast and humans. PMID- 15353336 TI - Prostaglandin E2 as a mediator of fever: the role of prostaglandin E (EP) receptors. AB - Prostaglandin E2 (PGE2) is a principal fever mediator that induces hyperthermia when injected into the organum vasculosum lamina terminalis (OVLT) and the adjacent preoptic area of the hypothalamus (POA). PGE (EP) receptors have four subtypes, i.e. EP1, EP2, EP3, and EP4. In the rat OVLT/POA region, at least three of these receptors, i.e. EP1, EP3, and EP4 receptors, have distinctively different distribution patterns. In rats, intracerebroventricular injection of EP1 receptor agonists and EP3 receptor agonists increased core temperature (Tc) and that of an EP4 receptor agonist decreased it. IntraPOA injection of an EP1 receptor agonist increased Tc. IntraPOA injection of an EP3 receptor agonist, however, induced hyperalgesia but not hyperthermia. Studies using mice with EP receptor gene deletions have indicated that EP3 receptors play a crucial role in febrile response. Therefore, the involvement of EP3 receptors at other levels of the nervous system should be considered. Such nuclei include the raphe pallidus nucleus, intermediolateral cell column in the spinal cord, or the nucleus of the solitary tract. PMID- 15353337 TI - Paradoxical effects of DNA binding polyamides on HTLV-1 transcription. AB - Human T-cell leukemia virus type-1 (HTLV-1) depends on the virally encoded transcription factor Tax for efficient viral replication and gene expression. In a complex with CREB, Tax contacts the minor groove of the promoter DNA at guanine and cytosine rich sequences that flank three of the off-consensus cyclic-AMP response elements (CREs). In this study, we used six Tax-directed pyrrole imidazole polyamides specifically designed to block Tax binding to DNA at each GC sequence of the three viral CREs. We found that four of these polyamides disrupt binding of the Tax/CREB complex in vitro, and that these same molecules also inhibit Tax-mediated transcription in vitro on chromatin-assembled templates. However, of these four Tax/CREB-specific polyamides, only one polyamide appears to be uniquely Tax specific. We show that polyamides can enter the nuclei of HTLV 1 infected T-cells, and two of the four polyamides down-regulated virion production in these cells. Together, these data illustrate the importance of studying polyamide inhibition of gene expression in vitro and in vivo, as the function of the polyamides in living cells is not fully understood. Finally, our data indicates that targeted disruption of the Tax/CREB complex, or other complexes which assemble on the HTLV-1 promoter, may provide a novel approach for inhibiting viral replication in vivo. PMID- 15353338 TI - Thermal homeostasis in systemic inflammation: modulation of neuronal mechanisms. AB - Fever can be defined as a specific model of body temperature control, modified by action of humoral substances released due to bacterial infection. Under laboratory conditions exogenous and endogenous pyrogens affect nervous endings in the body periphery, as well as thermosensitive neurons in the hypothalamus, which first manifests as a shock reaction and then as shifts of temperature thresholds for activation of thermoregulatory effectors (cold thermogenesis, vasomotion, sweating or panting) to higher body temperatures. During the later phase of fever, the temperature threshold for cold thermogenesis starts to move downwards, while the thresholds for other thermoregulatory outputs remain elevated, the result being enlarging of the interthreshold zone. This creates conditions for cooling of the body and for termination of the fever. During different phases of fever cytokines, prostaglandins, neuropeptides and catecholamines participate in modulation of mechanisms regulating thermoregulatory functions. This paper aims to specify the role of individual cytokines in induction of fever, as well as in activation of thermoregulatory centers as well as individual thermoregulatory effectors and to define differences in their mode of action. The paper further attempts to summarize our knowledge on humoral modulation of the cytokine release. It is concluded that cytokines are not the primary factors responsible for setting of the body thermostat during fever. PMID- 15353339 TI - Soluble interleukin-2 receptor in cancer. AB - Levels of serum sIL-2R (soluble interleukin-2 receptor) reflect the total amount of activated T lymphocytes in tumor infiltrating lymphocytes of cancer tissues and metastatic organs, because a part of alpha-chain of IL-2R is released into the bloodstream on the attachment of IL-2 (interleukin-2) to its specific IL-2R membrane. In most malignant diseases, elevated levels of serum sIL-2R are found, compare to normal control. Serum sIL-2R is a useful parameter for evaluating the disease stage and monitoring the disease progression during the post-treatment follow-up, though it is not an organ-specific parameter. PMID- 15353340 TI - Ginkgo biloba and neurodegenerative disorders. AB - The Gingko biloba extract EGb 761 has been the subject of many studies which confirm its usefulness for the prevention and treatment of neurodegenerative pathologies. These studies have focused on: a) the probable mechanisms of action that are involved in these disorders (including non-specific mechanisms implicated in diverse neurodegenerative disorders, particularly oxidative stress, or specific mechanisms such as those associated with beta-amyloid in Alzheimer's disease) and the processes of neuronal death; b) available animal models, and c) healthy individuals or those suffering from mild cognitive impairment or Alzheimer's disease. This data must be completed, particularly with regard to new knowledge about the pathogenesis of these disorders. Ambitious interventional studies are underway and may provide new evidence regarding the effect of EGb 761 in preventing Alzheimer's disease in humans. Positive findings would be particularly interesting since this drug is very safe to use. PMID- 15353341 TI - Angiogenesis and its role in the behavior of astrocytic brain tumors. AB - Angiogenesis, the development of new vessels from a pre-existing vasculature, accompanies the growth and malignant transformation of astrocytic brain tumors. Neovascularization is essential for sustained tumor growth, and with increasing grade, astrocytic tumors undergo an, angiogenic switch, manifested by marked increases in vessel density and changes in vascular morphology. In the quiescent state, endogenous anti-angiogenic factors including endostatin, thrombospondin, and soluble vascular endothelial growth factor receptor-1 (sVEGFR-1) balance the actions of pro-angiogenic stimuli and restrain the angiogenic switch. Once activated, pro-angiogenic factors including most notably basic fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF-A), and platelet-derived growth factor (PDGF) incite robust astrocytoma neovascularization. Recent studies have also explored the expression patterns and functional importance of the angiopoietins, Tie2 and neuropilin receptors, and hepatocyte growth factor/scatter factor (HGF). Together these angiogenic factors have diverse actions on endothelium and perivascular supporting cells that engender tumor neovessels with a unique phenotype, distinct from normal vessels. Properties of the astrocytoma neovasculature contribute to tumor growth, malignant progression, invasion, hemorrhage, and edema formation. Thus, the mechanistic actions of angiogenic factors on cerebral microvessels and the nature of the resultant tumor neovasculature establish a framework for understanding many of the characteristic behaviors of astrocytoma tumors. PMID- 15353342 TI - Cellular immunity and immunotherapy of brain tumors. AB - Factors influencing the host immune response to central nervous system (CNS) tumors are not yet well understood. This review will outline what is known about anti-tumor immune responses against CNS tumors and describe how advances in our knowledge of basic immunology may be applied to brain tumor immunotherapy. We will first focus on cellular immune system interactions involved in peripheral anti-tumor immune responses. Then, we will discuss characteristics of tumors arising within the confines of the CNS that distinguish them from peripheral neoplasms, emphasizing immune defects that seem to limit or curtail specific anti tumor immunity against brain tumors. Finally, the current state of immune-based treatment paradigms and future directions will be discussed, paying particular attention to adoptive cellular immunotherapy and tumor vaccine approaches for the treatment of malignant gliomas. PMID- 15353343 TI - Endothelial microparticles (EMP) bind and activate monocytes: elevated EMP monocyte conjugates in multiple sclerosis. AB - Elevated plasma endothelial microparticles (EMP) have been documented in MS during exacerbation. However, the role of EMP in pathogenesis of MS remains unclear. We investigated the formation of EMP-monocyte conjugates (EMP-MoC) and their potential role in transendothelial migration of inflammatory cells in MS. EMP-MoC were assayed in 30 MS patients in exacerbation, 20 in remission and in 35 controls. EMP-leukocyte conjugation was investigated flowcytometrically by employing alpha-CD54 or alpha-CD62E for EMP, and alpha-CD45 for leukocytes. EMP MoC were characterized by identifying adhesion molecules involved and their effect on monocyte function. In vivo (clinical): EMP-MoC were markedly elevated in exacerbation vs. remission and controls, correlating with presence of GD+ MRI lesions. Free CD54+ EMP were not elevated but free CD62E+ EMP were. In vitro: EMP bound preferentially to monocytes, less to neutrophils, but little to lymphocytes. Bound EMP activated monocytes: CD11b expression increased 50% and migration through cerebral endothelial cell layer increased 2.6-fold. Blockade of CD54 reduced binding by 80%. Most CD54+ EMP bound to monocytes, leaving little free EMP, while CD62+ EMP were found both free and bound. These results demonstrated that phenotypic subsets of EMP interacted differently with monocytes. Based on our observations, EMP may enhance inflammation and increase transendothelial migration of monocytes in MS by binding to and activating monocytes through CD54. EMP-MoC were markedly increased in MS patients in exacerbation compared to remission and may serve as a sensitive marker of MS disease activity. PMID- 15353344 TI - Mechanisms of alcohol liver damage: aldehydes, scavenger receptors, and autoimmunity. AB - While most of the investigations into the causative events in the development of alcoholic liver disease (ALD) have been focused on multiple factors, increasing interest has centered around the possible role of immune mechanisms in the pathogenesis and perpetuation of ALD. This is because many of the clinical features of ALD suggest that immune effector mechanisms may be contributing to liver tissue damage, as evidenced by the detection of circulating autoantibodies, and the presence of CD4+ and CD8+ lymphoid cells in the livers of patients with ALD. One mechanism that has been associated with the development of autoimmune responses is the modification (haptenation or adduction) of liver proteins with aldehydes or other products of oxidative stress. This is because it has been shown that these adducted proteins can induce specific immune responses, to the adduct, the adduct plus protein (conformational antigens), as well as the unmodified parts of the protein. More importantly, it is possible to demonstrate that adducted self-proteins can induce reactivity to the normal self-protein and thereby induce autoimmune responses. Therefore, it is the purpose of this manuscript to outline the mechanism(s) by which these modified self proteins can induce autoimmune reactivity, and thus play a role in the development and/or progression of ALD. PMID- 15353345 TI - Mechanisms of inflammation: the good, the bad and the ugly. AB - To the general public, the term, inflammation, is associated with pain, swelling, fever and a general sense of unease ranging from mere nuisance to debilitating illness. Under normal circumstances, the process of inflammation is actually a protective response designed to ward off invasion of the person by pathogens such as bacteria, viruses and/or parasites. The immune system of higher mammals (e.g. humans) is comprised of two distinct, arms, termed the innate and the adaptive systems. While these two components play unique roles in controlling pathogens, each relies, in some part, upon the effective function of the other in order to efficiently eliminate invading microorganisms. There are however situations in which this complex system is unable to properly function leading to unresolved infections and/or chronic states of inflammation. This review will summarize the basic mechanisms involved in the inflammatory process as well as discuss some of the key mediators and modulators of this process. PMID- 15353346 TI - N-nitrosations of basic amino acid residues in polypeptide. AB - Changes in the electrophoretic pattern were noted in the products of polypeptides of identical basic amino acids preincubated with reactive or degraded PN, suggesting the occurrence of N-nitrosation of the epsilon-amino group of lysine, the guanido group of arginine and the imidazole group of histidine. Additionally, increase in the N-nitroso immunoreactivity of preincubated histones H2A and H2B was detected by Western blot analysis. PMID- 15353347 TI - Metabolomic analysis of molecular species of phospholipids from normotensive and preeclamptic human placenta electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry (ESI/MS) is a highly sensitive, fast and powerful technique for the metabolite and metabolomic analysis of mixtures of lipids in a biological extract. We have exploited this technique to identify and characterize various phospholipids present in the placenta of preeclamptic and normal pregnant women. Multiple major molecular species can be detected in each phospholipid class have arachidonic acid as major fatty acid constituent. There is no remarkable difference in the molecular composition in each of these phospholipids in both the extracts. However, there seems to be lower amounts of the plasmenyl phosphatidylethanolamine and greater amounts of free fatty acids in preeclamptic placenta. PMID- 15353348 TI - Endogenous morphinergic signaling and tumor growth. AB - The mu3 opiate receptor subtype has been characterized by various binding assays as opiate alkaloid selective (e.g. morphine) and opioid peptide (e.g. methionine enkephalin) insensitive. This opiate receptor subtype has been found on human, including cancer cell lines, and invertebrate tissues, demonstrating that it has been conserved during evolution. Furthermore, in numerous reports, this receptor is coupled to constitutive nitric oxide release. In this regard, for example, morphine immune down regulating activities parallels those actions formerly attributed to nitric oxide. We have now identified the mu3 receptor at the molecular level and sequence analysis of the isolated cDNA suggests that it is a novel, alternatively spliced variant of the mu opiate receptor gene (MOR). Furthermore, using Northern blot, reverse transcription coupled to polymerase chain reaction (RT-PCR) and sequence analysis, we have demonstrated the expression of this new mu variant in human vascular tissue, mononuclear cells, polymorphonuclear cells, and human neuroblastoma cells. The presence of this mu splice variant, adds to the growing body of evidence supporting the hypothesis that morphine is an endogenous signaling molecule in neural, immune and vascular systems. In addition to their use in the treatment of pain, opioid peptides appear to be important in the growth regulation of normal and neoplastic tissue. This review will focus on the influence of opiate alkaloids, e.g., morphine, on tumor growth, with emphasis on immuno-regulatory and antiproliferative mechanisms. PMID- 15353349 TI - Human cell proteins and human immunodeficiency virus DNA integration. AB - Integration, catalyzed by the viral integrase (IN) protein, is a crucial step in the life cycle of all retroviruses including human immunodeficiency virus type 1 (HIV-1). Although purified HIV-1 IN protein is sufficient to catalyze the DNA breakage and joining steps of integration in the absence of any other protein factor, a number of studies indicate that cellular proteins participate in the integration process in cells. These host cell proteins have been proposed to act through binding the pre-integrated viral cDNA substrate, by directly interacting with the IN protein, and/or by repairing the single-stranded DNA gaps that occur at viral/chromosomal DNA junctions during integration. In this paper we summarize the identification and potential roles of specific cell factors in HIV-1 integration. We also present experimental results of human cell proteins that coimmunoprecipitated with HIV-1 IN following its expression in HeLa cells and discuss these results in light of the previously-identified integration cofactors. PMID- 15353350 TI - The role of tachykinins on bacterial infections. AB - Tachykinins represent a family of neuropeptides sharing similar C-terminus sequences, but exhibiting preferential binding to one of three receptors called neurokinin receptors (NK-R). While known for its role in contracting smooth muscle or acting as a pain signal neurotransmitter, substance P (SP) and other tachykinins can directly influence immune responses. Studies from the early 1980s revealed that human lymphocytes bore NK-R, but it remains unclear, even to-date, why such receptors are expressed on leukocytes. Nerve tracing studies have provided some speculation that the nervous system can assist the immune system in stimulating an immune response dependent upon which neuropeptide-bearing fibers infiltrate specific lymphoid structures. Such observations have important implications for regulating mucosal responses given that tachykinin-bearing nerve fibers extensively innervate the gut, and SP concentrations in the gut are second only to the brain. Such evidence suggests that SP and related neuropeptides may be important in controlling bacterial infections of the gut. This is shown by blocking SP action in which mice show increased susceptibility to Salmonella infections since induction of IFN-gamma is significantly reduced. In addition, the absence or its presence of SP's or the newly discovered lymphocyte-derived neurokinin called hemokinin's action can modify host IgA responses. Thus, tachykinins introduce new circuits to immune regulation suggesting that these neuropeptides exhibit cytokine- and chemokine-like action. PMID- 15353351 TI - HTLV envelopes and their receptor GLUT1, the ubiquitous glucose transporter: a new vision on HTLV infection? AB - We identified the ubiquitous glucose transporter GLUT1 as a receptor for Deltaretroviruses HTLV-1 and HTLV-2 envelopes (Env), mediating viral binding and entry. Here, we review the context and key observations that led us to this finding: functional modules of HTLV SU are similar to those of Gammaretrovirus Env which use multimembrane-spanning nutrient transporters as receptors; the HTLV Env receptor is an early marker of T lymphocyte activation; and HTLV Env inhibits glucose transport. We review several molecular, viral, cellular and physiological aspects of HTLV infection in relation to the in vivo and in vitro properties of GLUT1. Also, we examine the implications of HTLV-1 Env-GLUT1 interactions and altered glucose transport on the two major HTLV-1-induced diseases, adult T cell leukemia (ATL) and neurodegenerative tropical spastic paraparesis/HTLV-associated myelopathy (TSP/HAM). Complementary to the classical models of disease progression, we propose new schemes that emphasize the potential metabolic alterations caused in different cellular compartments. Finally, we review the potential use of HTLV Env-derived constructs as tools for labeling GLUT1 in vivo and inhibiting GLUT1 transport in tumor cells. PMID- 15353352 TI - IFN-beta in rheumatoid arthritis. AB - IFN-beta is a cytokine with pleiotropic effects and is expressed in rheumatoid synovial tissue. Based on in vitro work and experiments in animal models of rheumatoid arthritis (RA), the effects are mainly anti-inflammatory. Of special interest is the ability of IFN-beta to reduce the secretion of TNF-alpha, IL-1 beta, and IL-6, which are all key players in the pathogenesis of RA. At the same time IFN-beta could enhance the production of anti-inflammatory mediators like IL 1 receptor antagonist (IL-1Ra) and IL-10. Treatment of mice and monkeys with collagen-induced arthritis with daily IFN-beta injections resulted in clinical improvement, decreased synovial inflammation, and protection against joint destruction. Similar data were obtained after IFN-beta gene therapy. However, treatment of RA patients with IFN-beta has been unsuccessful so far, presumably due to pharmacokinetic issues. Novel approaches leading to constitutive IFN-beta production at the site of inflammation may be required to induce clinical efficacy in patients. PMID- 15353353 TI - A role for Pin1 in mammalian germ cell development and spermatogenesis. AB - The peptidyl-prolyl isomerase Pin1 is proposed to have diverse functions in many vital aspects of the cell. Despite the multitude of proteins targeted by Pin1 and the proposed regulatory role it plays in critical cellular functions, Pin1 is an essential gene in some eukaryotic organisms, but is dispensable in metazoans. In two genetic models, Candida albicans and Drosophila melanogaster, Pin1 participates in distinct developmental processes regulated by the MAPK pathway. Pin1-deficient mice exhibit decreased primordial germ cell proliferation during embryonic development, along with several degenerative or proliferative defects in the adult testis, retina, mammary gland, and brain. The combination of primordial germ cell deficit and spermatogonial depletion contributes to severe fertility defects in Pin1-null mice. Since growth factor activated MAPK pathways are vital to germ cell proliferation and differentiation, a role for Pin1 in mammalian germ cell development and spermatogenesis is discussed in the context of the Ras/MEK/MAPK pathway. PMID- 15353354 TI - Transglutaminase contributes to CPPD crystal formation in osteoarthritis. AB - Calcium pyrophosphate dihydrate (CPPD) crystals are common components of osteoarthritic joints and correlate with a poor prognosis. Transglutaminase (Tgase) enzymes have been implicated in pathologic mineralization in cartilage; yet, definitive studies linking Tgase activity to CPPD crystal formation in osteoarthritic articular cartilage are lacking. We measured in-vivo Tgase activity in osteoarthritic and normal human cartilage, and explored the effect of Tgase inhibitors on CPPD crystal formation by normal chondrocytes. Osteoarthritic articular cartilage from was obtained from specimens discarded at the time of knee replacement surgery. Normal adult cartilage samples from a tissue bank were used as controls. Tgase-specific isopeptide (epsilon-(gamma-glutamyl) lysine) bonds were measured in cartilage extracts by HPLC. Tgase-specific crosslinks were localized in osteoarthritic cartilage by immunohistochemistry. The effect of Tgase inhibition was determined in an in-vitro model of CPPD crystal formation. Tgase-specific crosslink levels were 1.55 +/- 0.3 picomoles/ng protein in normal human adult articular cartilage and 4.74 +/- 0.7 picomoles/ng protein in osteoarthritic human cartilage (p less than 0.001). Immunostaining confirmed the presence of Tgase crosslinks in the pericellular matrix of chondrocytes at potential sites of CPPD crystal formation. Tgase inhibitors suppressed CPPD crystal formation by porcine chondrocytes. These findings support a role for Tgase in CPPD crystal formation in aging or degenerated cartilage. PMID- 15353355 TI - A report on single exon genes (SEG) in eukaryotes. AB - Single exon genes (SEG) are archetypical of prokaryotes. Hence, their presence in intron-rich, multi-cellular eukaryotic genomes is perplexing. Consequently, a study on SEG origin and evolution is important. Towards this goal, we took the first initiative of identifying and counting SEG in nine completely sequenced eukaryotic organisms--four of which are unicellular (E. cuniculi, S. cerevisiae, S. pombe, P. falciparum) and five of which are multi-cellular (C. elegans, A. thaliana, D. melanogaster, M. musculus, H. sapiens). This exercise enabled us to compare their proportion in unicellular and multi-cellular genomes. The comparison suggests that the SEG fraction decreases with gene count (r = -0.80) and increases with gene density (r = 0.88) in these genomes. We also examined the distribution patterns of their protein lengths in different genomes. PMID- 15353356 TI - Susceptibility and resistance to experimental adjuvant arthritis. AB - Autoimmunity is the result of an abnormal immune response against constituents of body tissues. For many years, the study of animal models of human diseases was aimed at defining the factors participating in the autoimmune process. During the past two decades, much of the attention was diverted to another intriguing aspect of animal models: the mechanisms rendering some animal strains autoimmune susceptible and others resistant. In this report, we focus on one experimental model, adjuvant arthritis (AA) which is inducible in the Lewis rat following stimulation of the immune system by heat-killed mycobacterium and its 65kDa heat shock protein. We describe genetic loci regulating the severity of this disease as well as the contribution of microbial flora and endocrine activity to susceptibility and resistance. In our opinion, a better understanding of the processes underlying susceptibility and resistance to AA is an important step towards the development of new therapeutic approaches to autoimmunity. PMID- 15353357 TI - Molecular and biological effects of hemodynamics on vascular cells. AB - A variety of systemic risk factors, including smoking, hypertension, hyperlipidemia and diabetes have been found to promote atherosclerosis. Although these elements affect blood vessels equally, clinically significant lesions develop at predictable locations, i.e., major branch points and bifurcations. This suggests that the development of clinically significant atherosclerotic plaques involves a complex interplay between vascular anatomy, vascular biology and hemodynamic forces. Cyclic strain, circumferential pulsatile pressure exerted upon a vessel wall, has been found to cause changes in endothelial cells that tend to disfavor atherosclerosis formation. Cultured endothelial cells have been shown to migrate, proliferate and alter cytoskeletal alignment in response to cyclic strain. Levels of macromolecules such as prostacyclin, endothelin, nitric oxide and tissue plasminogen activator have been found to be altered by cyclic strain. Additionally, cyclic strain has been shown to stimulate expression of cellular adhesion molecules such as ICAM-1 and intracellular second messenger systems such as the adenylate cyclase-cAMP, diacylglycerol-IP3, and protein kinase C pathways. This article reviews the most current pertinent literature and summarizes the presently known effects of cyclic strain on endothelial cells. PMID- 15353358 TI - An unconventional hypothesis of oxidation in Alzheimer's disease: intersections with excitotoxicity. AB - There are two major lines of investigation from which a connection has been traditionally drawn between chemical oxidation and Alzheimer's disease. First, a major risk factor for AD is age, and oxidative stress has long been a component of general hypotheses about biological aging. The second line of reasoning is a corollary of the Amyloid Hypothesis, the assumption that the amyloid beta-peptide (A-beta) which comprises AD's pathognomic plaques is a key mediator of the neurodegeneration occurring in this disorder. Under many experimental conditions, A-beta has been shown to evoke oxidative damage to tissues, cells, and biomolecules; even the redox properties of the peptide itself have been hotly debated. These two modalities of conjecture intersect under the Inflammatory Hypothesis of AD, as inflammation produces oxidation, old age is associated with elevation in inflammatory events, and A-beta can further exacerbate such inflammatory reactions in brain cells. This review discusses these arguments about the pathogenesis of AD and how they might be generalized to other neurodegenerative conditions. But, additional speculation is offered in the form of an inclusionary mechanism that may be specific and novel enough to qualify as a third line of theory; namely, the possibility that inflammatory reactions in microglia--activated by A-beta or other factors among the "usual suspects"- initiate programmed oxidation that is converted to the neuron-specific stress of excitotoxicity. PMID- 15353359 TI - The protein folding problem: global optimization of the force fields. AB - The evolutionary development of a theoretical approach to the protein folding problem, in our laboratory, is traced. The theoretical foundations and the development of a suitable empirical all-atom potential energy function and a global optimization search are examined. Whereas the all-atom approach has thus far succeeded for relatively small molecules and for alpha-helical proteins containing up to 46 residues, it has been necessary to develop a hierarchical approach to treat larger proteins. In the hierarchical approach to single- and multiple-chain proteins, global optimization is carried out for a simplified united residue (UNRES) description of a polypeptide chain to locate the region in which the global minimum lies. Conversion of the UNRES structures in this region to all-atom structures is followed by a local search in this region. The performance of this approach in successive CASP blind tests for predicting protein structure by an ab initio physics-based method is described. Finally, a recent attempt to compute a folding pathway is discussed. PMID- 15353360 TI - Porous carbon composite/enzyme glucose microsensor. AB - An enzyme glucose microsensor using a glucose oxidase-immobilized porous carbon/Teflon composite microelectrode was developed. The microsensor was fabricated by etching a platinum microelectrode (platinum, radius of 25 and 50 micrometers) in hot aqua regia to create a cavity at the tip and then packing a porous carbon/Teflon composite, which was made from acetylene black and Teflon emulsion, into the cavity. Nafion was impregnated into the inner surface of porous carbon/Teflon composite electrode following immobilization of Os(bpy)3+2/+3 as electron transfer mediators. The loading amount of Os(bpy)3+2/+3 in the Nafion/porous carbon/Teflon composite electrode was found to be 7.0x10(-8) mole cm(-2), which is much higher than that in polymer modified electrodes reported in literatures. The microsensor was further dipped overnight in buffer solution containing glucose oxidase for enzyme modification. With both glucose oxidase and mediators in the porous carbon/Teflon composite surface, the sensor performance was evaluated in buffer solutions containing different glucose concentrations and serum samples for glucose determination. The microsensor showed directly electrochemical glucose oxidation on the Os(bpy)3+2/+3 impregnated enzyme/porous carbon/Teflon composite surface with linear response over concentration range of 0-15 mM and Machaelis behavior. Reliability and reproducibility were conducted in serum samples and glucose buffer solution, and the results demonstrated there was no significant decrease of amperometric response in air-saturated solution for one month. The sensor demonstrated potential in clinical diagnostic applications. PMID- 15353361 TI - Testing the amyloid toxicity hypothesis of Alzheimer's disease in transgenic Caenorhabditis elegans model. AB - Alzheimer's disease (AD) is affecting more people every year due to the increase in elderly population. This disease is characterized by senior plaques, containing aggregated amyloid beta peptide (A beta), and neurofibrillary tangles in the AD brains. The A beta depositions are thought to increase in cellular oxidative stress, which subsequently produces neuronal cell death in the patient s brain, causing loss of memory and, in the latter stages, dementia. Diverse models have been established to test this, "Amyloid Toxicity Hypothesis of AD". Among these, the use of the nematode Caenorhabditis elegans has some advantages. This invertebrate has its entire genome known, as well as numerous gene homologues to those seen in humans. In relationship with the cell model, the nematode gives the benefit of an organismal view of the disease. The nematode's short life span proves useful, when compared with that of mice, allowing mechanistic studies of the disease and pharmacological treatments. Alongside with other laboratories, we have used this in vivo model to correlate the Abeta expression with its toxicity through the observance of the organism's behavior to provide a better understanding of the cellular processes underlining AD. PMID- 15353362 TI - Non-prostaglandin eicosanoids in fever and anapyrexia. AB - Until recently, studies on the role of the metabolites of arachidonic acid (AA), eicosanoids in fever have primarily focused on prostaglandins, prostaglandin E2 (PGE2) in particular, derived from the pathway related to cyclooxygenases (COX). COX exists in two known isoforms; a constitutive COX-1, and COX-2, which is inducible upon the action of pyrogens. Data accumulated in our laboratories suggest a thermoregulatory role for two other pathways of arachidonate metabolism; 5-lipoxygenase (5-LOX) and cytochrome P-450 (epoxygenase). We have demonstrated that leukotrienes (LTs; 5-LOX-derived eicosanoids) and various isomers of epoxyeicosatrienoic acids (EETs; epoxygenase-derived eicosanoids) contribute to the process of endogenous antipyresis or cryogenesis, which limits the height of fever. In support of this are several lines of evidence based on both in vivo and in vitro experiments. 1) Intracerebroventricular (icv) injections of LTC4 at nanomolar concentrations cause a dose-dependent decrease of body temperature (Tb) in mice. 2) Lipopolysaccharide (LPS)-induced anapyrexia in mice is preceded and accompanied by elevation in hypothalamic cysteinyl-LT (CysLT) production. 3) The inhibitor of LT synthesis MK-886 suppresses both of these processes. 4) EETs as well as inducers of the epoxygenase attenuate, whereas inhibitors of epoxygenase enhance the LPS-induced fever in rats. 5) One of the isomers of EET, 11,12-EET, in in vitro studies inhibited both the generation of PGE2 and IL-6 in monocytes stimulated with LPS. These results, together with a well-established pyrogenic role of PGE2, indicate that AA cascade may be regarded as an endogenous system to regulate the temperature response upon disease. COX, 5-LOX, and epoxygenase products may act at the level of hypothalamus as proximal mediators of, respectively, fever (PGE2) or cryogenesis (CysLTs and EETs), or indirectly by influencing the other endogenous cryogens and pyrogens. PMID- 15353363 TI - Plasminogen activator/plasminogen activator inhibitors in ovarian physiology. AB - The target extracellular matrix (ECM) degradation generated by plasminogen activator (PA) and regulated by plasminogen activator inhibitor (PAI) is an event that affects a wide variety of physiological and pathological processes in the ovary. Studies carried out over the past 25 years in a number of laboratories have elucidated some of the biochemical events related to the function and regulation of the PA system in the ovary. Hormone-induced coordinated expression of tissue-type PA (tPA) produced mainly by granulosa cells and its inhibitor PAI 1 secreted by theca cells in the preovulatory follicles is responsible for a controlled and directed proteolysis leading to the rupture of selected follicles in the rat, monkey and other mammals. Increase in tPA and PAI-1 expression in corpus luteum (CL) of rat and monkey at a later stage is well correlated with a sharp decrease in CL progesterone production, indicating its important role in the initiation of luteal regression. In contrast, the urokinase-type PA (uPA) may play an essential role in the early growing follicles during cell proliferation and migration, and in the early CL formation related to ECM degradation and angiogenesis. Ovarian function is also modulated by endogenously-produced local factors that regulate expression of the PA activator and inhibitor, and the MMP system. Thus, the next challenge is to identify the interrelationship between multiple paracrine and autocrine factors and the PA system, and to know how they regulate the protease and the protease inhibitor in the ovary. PMID- 15353364 TI - Genetic immune modulation of Ran GTPase against different microbial pathogens. AB - Septic shock characterized by pro-inflammatory cytokine storm can be induced by a variety of microbial infections. Typical pro-inflammatory cytokines include TNFalpha, IL1 and IL6. Although one or more of them is often expressed in any given microbial infection, usually it is the elevation of one cytokine that becomes predominant at a particular time in a given infection. Here we showed that administration of adenoviral antigens alone led to a predominant elevation of serum IL6 but not TNFalpha Administration of endotoxin together with adenoviral antigens led to elevation of both serum IL6 and TNFalpha. In vivo expression of RanC/d, but not RanT/n or LacZ into peritoneal macrophages rapidly down-modulated the levels of these cytokines in both experimental situations. It also correlated with reduced liver inflammatory damage and increased resistance to septic shock. We conclude that RanC/d can be applied to down-modulating production of cytokines induced by microbial products other than endotoxin and to render resistance to mice against septic shock induced by one or more microbial pathogens. The ability of using RanC/d to down-modulate and RanT/n to up regulate host innate immune response induced by multiple microbial pathogens is illustrated in this study. Incorporation of either or both RanC/d and RanT/n alleles into appropriate vectors will produce genetics vaccines valuable for biodefense and medically important illness in which host immune system against invading agents is severely burdened. PMID- 15353365 TI - Lactate biosensors for continuous monitoring. AB - Blood lactate concentration is a highly sensitive measure of tissue oxygen deprivation from ischemia, trauma, and hemorrhage, which can produce life threatening shock. Significantly, blood lactate is the most reliable end point indicator of resuscitation and predictor of survivability. The need for continuous measurement of blood lactate, and the slowness of discrete conventional photometric assays, is leading to the development of monitoring systems based on electrochemical lactate biosensors. Research and development of both in vivo and ex vivo biosensor systems is ongoing. Ex vivo biosensors are used with implanted microdialysis or ultrafiltration probes. Lactate from the blood diffuses into the dialysate/filtrate is transported outside of the body for measurement. The external biosensors are protected from fouling or contamination from unwanted blood constituents, but at the cost of an inherent delay in response despite system miniaturization. In vivo biosensors provide a direct measurement of blood lactate concentration, providing rapid response to changes in lactate levels. In vivo sensors are placed in the skin or implanted subcutaneously. Response to changes in lactate concentration is rapid, but biocompatibility requirements are more stringent than for ex vivo sensors. As is the case with all microdialysis systems, some in vivo biosensors must be implanted into the body using an insertion needle or surgical means, limiting their use. However, small, inexpensive, disposable in vivo sensors are also being developed which can emplaced and removed by the user. PMID- 15353366 TI - Delta opioid peptide (D-Ala 2, D-Leu 5) enkephalin: linking hibernation and neuroprotection. AB - Hibernation is a potential protective strategy for the peripheral, as well as for the central nervous system. A protein factor termed hibernation induction trigger (HIT) was found to induce hibernation in summer-active ground squirrels. Purification of HIT yielded an 88-kD peptide that is enriched in winter hibernators. Partial sequence of the 88-kD protein indicates that it may be related to the inhibitor of metalloproteinase. Using opioid receptor antagonists to elucidate the mechanisms of HIT, it was found that HIT targeted the delta opioid receptors. Indeed, delta opioid (D-Ala 2, D-Leu 5) enkephalin (DADLE) was shown to induce hibernation. Specifically, HIT and DADLE were found to prolong survival of peripheral organs, such as the lung, the heart, liver, and kidney preserved en bloc or as a single preparation. In addition, DADLE has been recently demonstrated to promote survival of neurons in the central nervous system. Exposure to DADLE dose-dependently enhanced cell viability of cultured primary rat fetal dopaminergic cells. Subsequent transplantation of these DADLE treated dopaminergic cells into the Parkinsonian rat brain resulted in a two-fold increase in surviving grafted cells. Interestingly, delivery of DADLE alone protected against dopaminergic depletion in a rodent model of Parkinson s disease. Similarly, DADLE blocked and reversed the dopaminergic terminal damage induced by methamphetamine (METH). Such neuroprotective effects of DADLE against METH neurotoxicity was accompanied by attenuation of mRNA expressions of a tumor necrosis factor p53 and an immediate early gene c-fos. In parallel to these beneficial effects of DADLE on the dopaminergic system, DADLE also ameliorated the neuronal damage induced by ischemia-reperfusion following a transient middle cerebral artery occlusion. In vitro replication of this ischemia cell death by serum-deprivation of PC12 cells revealed that DADLE exerted neuroprotection in a naltrexone-sensitive manner. These results taken together suggest that DADLE stands as a novel therapeutic agent. In this review paper, we present laboratory evidence supporting the use of DADLE for protection of peripheral and central nervous system. PMID- 15353367 TI - Virulence factors of the group A streptococci and genes that regulate their expression. AB - Group A streptococci produce a wide variety of extracellular (cell-associated and released) virulence factors. The function of these factors varies and includes roles in adhesion, spreading, tissue destruction, immune system evasion, and cell toxicity. How these factors are regulated with regard to one another is important for this organism s ability to bring about the variety of diseases this microbe is capable of causing. Therefore, along with the multitude of virulence factors, there are a number of regulatory systems that regulate, either directly or indirectly, the production of these factors and therefore influence the pathogenesis of group A streptococcal infections. PMID- 15353368 TI - Real time and in vivo monitoring of nitric oxide by electrochemical sensors--from dream to reality. AB - Nitric oxide is a key intercellular messenger in the human and animal bodies. The identification of nitric oxide (NO) as the endothelium-derived relaxing factor (EDRF) has driven an enormous effort to further elucidate the chemistry, biology and therapeutic actions of this important molecule. It has found that nitric oxide is involved in many disease states such as such as chronic heart failure, stroke, impotent (erectile dysfunction). The bioactivity of nitric oxide intrinsically linked to its diffusion from its site production to the sites of action. Accurate reliable in real time detection of NO in various biological systems is therefore crucial to understanding its biological role. However, the instability of NO in aqueous solution and its high reactivity with other molecules can cause difficulties for its measurement depending on the detection method employed. Although a variety of methods have been described to measure NO in aqueous environments, it is now generally accepted that electrochemical (amperometric) detection using NO-specific electrodes is the most reliable and sensitive technique available for real-time in situ detection of NO. In 1992 the first commercial NO electrode-based amperometric detection system was developed by WPI. The system has been used successfully for a number of years in a wide range of research applications, both in vitro and in vivo. Recently, many new electrochemical nitric sensors have been invented and commercialized. Here we describe some of the background principles in NO sensors design, methodology and their applications. PMID- 15353369 TI - Natural antioxidants and neurodegenerative diseases. AB - Neurodegenerative diseases such as Alzheimer's (AD) and Parkinson's (PD) diseases are defined by a progressive neuronal dysfunction and an ensuing behavioral dysfunction. Although protein aggregation (i.e beta-amyloid and alpha-synuclein) plays a pivotal role in both AD and PD, there is increasing evidence that excessive accumulation of reactive oxygen species (ROS) that occurs during normal and pathological brain aging contributes to neuronal losses and dysfunction. Based on these observations, it has been hypothesized that natural antioxidants derived from food, beverages and natural extracts may be beneficial to prevent or delay the occurrence of age-related cognitive deficits and neurodegenerative diseases. We will summarize in this review the role of oxidative stress in pathological brain aging, and provide evidence for a role for antioxidant molecules as therapeutic agents. We will also focus on the various mechanisms underlying their neuroprotective effects in in vivo and in vitro models of neurotoxicity. PMID- 15353370 TI - Insights into the catalytic mechanism of peptidyl prolyl cis/trans isomerases. AB - A large body of physiological, cell biological, kinetic and structural data about peptidyl prolyl cis/trans isomerases (PPIases) has been accumulated during the past 20 years, but despite the simplicity of the catalyzed reaction the question of how the enzyme action is performed is still not fully answered. In this review the center of attention is the molecular background of the catalytic mechanism of PPIases and the spontaneously occurring peptidyl prolyl cis/trans isomerization. We summarize and compare the available kinetic, structural and amino acid sequence data of all three PPIase families, the cyclophilins, FKBP and parvulins. Different catalytic mechanisms that have been suggested in the literature are discussed. A comprehensive comparison of enzyme active site structures reveals a hitherto unnoticed similarity between the three PPIase families and might suggest that PPIases utilize mechanisms that are more similar than previously suspected. PMID- 15353371 TI - Porous carbon composite/enzyme glucose microsensor. AB - An enzyme glucose microsensor using a glucose oxidase-immobilized porous carbon/Teflon composite microelectrode was developed. The microsensor was fabricated by etching a platinum microelectrode (platinum, radius of 25 and 50 micrometers) in hot aqua regia to create a cavity at the tip and then packing a porous carbon/Teflon composite, which was made from acetylene black and Teflon emulsion, into the cavity. Nafion was impregnated into the inner surface of porous carbon/Teflon composite electrode following immobilization of Os(bpy)3+2/+3 as electron transfer mediators. The loading amount of Os(bpy)3+2/+3 in the Nafion/porous carbon/Teflon composite electrode was found to be 7.0x10(-8) mole cm(-2), which is much higher than that in polymer modified electrodes reported in literatures. The microsensor was further dipped overnight in buffer solution containing glucose oxidase for enzyme modification. With both glucose oxidase and mediators in the porous carbon/Teflon composite surface, the sensor performance was evaluated in buffer solutions containing different glucose concentrations and serum samples for glucose determination. The microsensor showed directly electrochemical glucose oxidation on the Os(bpy)3+2/+3 impregnated enzyme/porous carbon/Teflon composite surface with linear response over concentration range of 0-15 mM and Machaelis behavior. Reliability and reproducibility were conducted in serum samples and glucose buffer solution, and the results demonstrated there was no significant decrease of amperometric response in air-saturated solution for one month. The sensor demonstrated potential in clinical diagnostic applications. PMID- 15353372 TI - Development of new anti-cancer peptides from conformational energy analysis of the oncogenic ras-p21 protein and its complexes with target proteins. AB - We have employed a computational approach to design peptides, from known oncogenic proteins, that inhibit tumor growth. This approach has been applied to the ras-p21 protein that becomes oncogenic when single amino acid substitutions occur at critical positions in its polypeptide chain, such as at Gly 12 and Gln 61. In this approach, using two sampling methods, molecular dynamics and the electrostatically driven Monte Carlo (EDMC) method, we have computed the average structures of wild-type and oncogenic forms of ras-p21 alone and bound to a number of its target proteins, such as the ras-binding domain (RBD) of raf, guanine nucleotide exchange protein (GAP) and SOS guanine nucleotide exchange protein (GAP). By superimposing the average structures of the oncogenic forms on those of their wild-type counterparts, we have identified a number of domains that change conformation. These domains are potential effector domains that are involved uniquely in oncogenic ras-p21 signaling. We have therefore synthesized peptides corresponding to these domains and tested them in Xenopus laevis oocytes for their abilities to inhibit oncogenic ras-p21 selectively. Using this approach, we have identified three peptides from ras-p21 and one peptide each from the RBD of raf, GAP and SOS that selectively inhibit oncogenic but not insulin-activated wild-type ras-p21-induced oocyte maturation. We have synthesized the ras-p21 peptides attached to a penetratin leader sequence to enable cell membrane penetration and introduced these peptides into a ras transformed pancreatic cancer cell line; these peptides, but not an unrelated negative control peptide, cause the cells to undergo complete phenotypic reversion. On the other hand, none of these peptides has any effect on the growth of untransformed pancreatic acinar cells in culture, further suggesting that they may not interfere with normal cell growth. Thus these peptides can be useful agents in the treatment of cancers. We have further used these peptides to demonstrate that oncogenic and wild-type ras-p21 proteins utilize different signal transduction pathways and to identify where these differences occur in cells. PMID- 15353373 TI - Opioid peptides in cerebrospinal fluid-methods for analysis and their significance in the clinical perspective. AB - The discovery of the endogenous opioid peptide systems and their subsequent identification in human cerebrospinal fluid near 30 years ago triggered an intensive research to evaluate the function of these compounds in the clinical perspective. However, for this purpose it was necessary to develop reliable techniques with high sensitivity and reproducibility. Furthermore, it was necessary to assess the chemical nature of the opioid activity present in CSF. Therefore, research on opioid peptides in CSF have to a considerable extent been directed to attempts to characterize the peptide activity present in this fluid in order to identify suitable markers of activity in any particular opioid peptide system. In the clinic these markers have been used in attempts to correlate alterations in peptide levels to various neurological diseases. This article reviews the past and ongoing research on opioid peptide systems in CSF from human with particular emphasis on their relevance in the clinical perspective. PMID- 15353374 TI - Trauma symptoms, sexual behaviors, and substance abuse: correlates of childhood sexual abuse and HIV risks among men who have sex with men. AB - Childhood sexual abuse is associated with high-risk sexual behavior in men who have sex with men. This study examined psychological and behavioral correlates of HIV risk behavior associated with childhood sexual abuse in a sample of men who have sex with men. Men attending a large gay pride event (N = 647) completed anonymous surveys that assessed demographic characteristics, childhood sexual abuse history, symptoms of dissociation and trauma-related anxiety, borderline personality characteristics, substance use, and sexual risk behavior. Results indicated that men who have a history of childhood sexual abuse were more likely to: engage in high-risk sexual behavior (i.e., unprotected receptive anal intercourse), trade sex for money or drugs, report being HIV positive, and experience non-sexual relationship violence. Results of this study extend previous research to show that men who have sex with men and who have a history of child sexual abuse are more likely to be at high risk for HIV infection. PMID- 15353375 TI - Childhood sexual and physical abuse and adult vulnerability to PTSD: the mediating effects of attachment and dissociation. AB - Two hundred and eighty-four adults from the metropolitan New York area reported on their history of childhood sexual abuse (CSA), childhood physical abuse (CPA), and on the nature of their exposure to the terrorist attack on the World Trade Center. The respondents also completed the Impact of Events Scale-Revised (IES R), the Attachment Style Questionnaire (ASQ) and the Dissociative Experiences Scale (DES). Those reporting histories of CSA and/or CPA were found to endorse more serious symptoms of PTSD, as did those who witnessed the terrorist attack live. The presence of secure attachments and dissociative symptoms were related significantly to both CSA and CPA, and to scores on the IES-R. Adult attachment and dissociation were found to mediate the relationship between childhood abuse and severity of PTSD. PMID- 15353376 TI - Measuring child sexual abuse in children and youth. AB - Asking children and adolescents directly about their experience of sexual victimization overcomes some of the methodological weaknesses inherent in other approaches. Yet complex legal, ethical, and methodological issues remain. This paper reviews the psychometric properties of those questions or instruments that have measured exposure to child sexual abuse directly. A search of four electronic databases using descriptors "child sexual abuse" and "measurement" or "instrumentation" yielded four telephone administered tools, 13 face-to-face interviews, and 32 self-administered questionnaires. Few instruments had been subjected to rigorous evaluation. Establishing the validity and reliability of instruments measuring child sexual abuse and other forms of victimization are critical for the growth and expansion of the field. PMID- 15353377 TI - Assessing the long-term effects of EMDR: results from an 18-month follow-up study with adult female survivors of CSA. AB - This 18-month follow-up study builds on the findings of a randomized experimental evaluation that found qualified support for the short-term effectiveness of Eye Movement Desensitization and Reprocessing (EMDR) in reducing trauma symptoms among adult female survivors of childhood sexual abuse (CSA). The current study provides preliminary evidence that the therapeutic benefits of EMDR for adult female survivors of CSA can be maintained over an 18-month period. Furthermore, there is some support for the suggestion that EMDR did so more efficiently and provided a greater sense of trauma resolution than did routine individual therapy. PMID- 15353378 TI - A videotape series for teaching physicians to evaluate sexually abused children. AB - A free videotape subscription series was utilized to increase the knowledge of general physicians in clinical practice about the medical evaluation of sexually abused children. Of the 65 physicians who requested the first tape, 39 (60%) completed it. Fourteen of the 39 physicians who completed the first tape (36%) completed the 5-tape series. Completion data suggested that series completion was unrelated to prior knowledge, years since training or number of sexual abuse examinations performed in the previous year. Evaluative comments suggested that quality of the tapes was not a factor in completion rate. On tests of immediate retention, the average posttest percent correct was significantly higher than on the pretest. In a 3-year follow-up of the 14 physicians who completed the series, 10 reported that they were still performing sexual abuse examinations. PMID- 15353379 TI - Involvement in high-profile child sexual abuse controversies: costs and benefits. AB - The costs and benefits associated with participation in conflicts around child sexual abuse are explored. Using qualitative data from interviews with 40 high profile participants, findings suggest that professionals experience both positive and negative consequences as a result of their involvement. Costs identified include attacks and misrepresentations, harm to health or emotional well-being, and career losses or changes. Benefits included personal growth, a sense of personal satisfaction, and career enhancement. Women were found to have experienced more impacts overall than men. Psychological theories about stress and coping and sociological theories about social movement participation are used to explore why people remain committed despite the risks associated with participation. Material rewards, commitment to change, the development of a highly salient identity, and the support of social networks may be contributing factors. PMID- 15353381 TI - Is Alzheimer disease a mental disorder? PMID- 15353383 TI - Alzheimer disease with psychosis: excess cognitive impairment is restricted to the misidentification subtype. AB - OBJECTIVE: Psychotic symptoms occur in 30%-60% of individuals with Alzheimer disease (AD) with psychosis (AD+P). AD+P identifies a distinct AD phenotype, with increased severity of cognitive impairment and a more rapid cognitive decline. Using factor and cluster analysis, we previously proposed two subtypes of patients with AD+P, one characterized by misidentifications and hallucinations (Misidentification), the other by persecutory delusions (Paranoid). We hypothesized that these two groups differed in their patterns of cognitive impairment, compared with AD subjects without psychosis. METHODS: Subjects (N=119) with possible or probable AD were assessed with a comprehensive neuropsychological test battery at the time of initial presentation. Psychotic symptoms were ascertained with the CERAD Behavioral Rating Scale. Cognitive test scores were compared among groups by use of general linear-regression models, with age, education, and duration of illness entered as covariates. All results were corrected for multiple comparisons. RESULTS: The Misidentification group was significantly more impaired than the Non-Psychotic group on tests of verbal fluency and visuospatial function. The Paranoid group did not differ from the Non Psychotic group on any test. CONCLUSIONS: These results support the identification of the Misidentification and Paranoid groups as distinct subgroups of AD+P. The ability to detect meaningful biologic associations of AD+P in future studies would be enhanced by separate analysis of the Misidentification and Paranoid phenotypes. PMID- 15353384 TI - End-of-life care: issues relevant to the geriatric psychiatrist. AB - Most deaths in the United States occur in the context of chronic diseases in later life and are too often accompanied by potentially remediable emotional or physical suffering. Geriatric psychiatrists and other mental health professionals can contribute meaningfully to the provision of optimal care during the final phases of life. This review provides an overview of end-of-life care, focusing on issues most relevant to the geriatric psychiatrist. The author examined palliative care textbooks and review papers to determine the topics to be included in this article, and searched computerized literature databases on these topics. Many of the recommendations provided herein stem from experts' clinical experience; however empirical evidence is also incorporated and critiqued. Topics covered include conversations with patients and families about end-of-life care; the evaluation and treatment of suffering, including pain, depression, suicidality, anxiety, and delirium; the role of individual and family therapies in caring for dying patients; capacity determination; advance care planning; withholding life-sustaining treatments; and "last resort" (and, in some cases, quite controversial) options, such as terminal sedation, assisted suicide, and euthanasia. The author also notes the relevance of such end-of-life-care considerations to patients with dementia. Geriatric psychiatrists' skills across these multiple domains are of particular usefulness. Through such clinical skills and the application of empirical research tools to the many unanswered questions in the care of dying patients, geriatric psychiatry can make increasingly valuable and visible contributions to improving quality of life for people suffering from life-threatening illnesses. PMID- 15353385 TI - Long-term outcomes of galantamine treatment in patients with Alzheimer disease. AB - OBJECTIVE: The authors evaluated the long-term safety, efficacy, and tolerability of galantamine 24 mg/day in the treatment of Alzheimer disease by means of a 12 month, open-label extension of an earlier 5-month, double-blind, placebo controlled trial with a 6-week withdrawal phase. METHODS: Patients completing two double-blind, placebo-controlled trials (N=699) were escalated to a 24-mg dose (12 mg bid) of galantamine during a period of 2 weeks and treated for 12 months beyond the initial 6.5-month, double-blind period (total treatment duration: 18.5 months). The primary efficacy measure was the change from baseline in the Alzheimer's Disease Assessment Scale (ADAS-Cog/11) score at 18.5 months; secondary endpoints included total scores on the Alzheimer's Disease Cooperative Study of Activities of Daily Living and the Neuropsychiatric Inventory. Standard safety evaluations, including adverse-event monitoring, were performed. RESULTS: Patients taking galantamine continuously throughout the double-blind and open label studies (N=288) showed sustained cognitive benefits on ADAS-Cog/11 scores at 18.5 months. Patients were maintained close to baseline cognitive ability for 12 months, and safety was as expected and documented in other large studies of galantamine. Analysis of the subgroup of patients (N=113) who completed the entire 18.5 months of galantamine treatment showed that cognitive function was maintained up to 14 months. CONCLUSIONS: Results of this open-label extension support the findings from previous galantamine studies and demonstrate the safety and tolerability of galantamine for up to 18.5 months. PMID- 15353386 TI - Effective recruitment strategies for Latino and Caucasian dementia family caregivers in intervention research. AB - OBJECTIVE: Latino family caregivers of older people with memory problems are a rapidly growing group. However, their participation in intervention research is disproportionately low. The authors compared the participant retention rates of three different recruitment strategies in a sample of Latino and Caucasian family caregivers. METHODS: A total of 310 caregivers (195 Latino and 105 Caucasian) were screened for participation in one of two clinical intervention trials. Potential participants were recruited from one of three sources: 1) non professional (e.g., health fairs); 2) professional (e.g., community agencies); and 3) advertisements. Reasons for non-participation were also obtained. RESULTS: Caucasians were significantly more likely to be successfully retained across all recruitment strategies (52% versus 31%, respectively). However, logistic regression revealed a recruitment strategy-by-ethnicity interaction: Latinos were highly similar in retention rates to Caucasians when recruited through professional referrals, and far less likely to participate with other strategies. Descriptive analyses also revealed ethnic differences for non-participation. CONCLUSION: Partnership of Latino family caregivers with community agencies may result in the highest recruitment rates among this group. Latino cultural values likely play a role in the success of these partnerships in participant recruitment. PMID- 15353387 TI - Cognitive response to pharmacological treatment for depression in Alzheimer disease: secondary outcomes from the depression in Alzheimer's disease study (DIADS). AB - OBJECTIVE: The authors assessed the cognitive effects of depression treatment with sertraline in patients with Alzheimer disease (AD) and major depression. METHODS: Forty-four patients with probable AD and major depression were enrolled in a double-blind, placebo-controlled clinical trial of sertraline. Cognitive testing was done at baseline and at 3-week intervals throughout the 12-week study. At the 12th week, subjects were categorized by treatment response (full, partial, or no response). Cognitive data from 41 participants who completed three or more testing sessions and 36 who completed all five study visits were included in the analyses. RESULTS: Neither improved mood nor use of sertraline was associated with cognitive change over time in AD patients. Post-hoc exploration of the data, however, suggested a sex difference in cognitive response to sertraline such that women treated with sertraline demonstrated improved cognition compared with women on placebo, whereas men treated with sertraline worsened significantly in cognition compared with men on placebo. CONCLUSIONS: In this study, among depressed AD patients after treatment with sertraline or placebo, there was no evidence that improved mood was associated with cognitive improvement. Future studies aimed at increasing power to detect mood as well as medication effects will be valuable in determining the relationship between cognition and treatment of depression in AD patients. PMID- 15353388 TI - Risperidone and falls in ambulatory nursing home residents with dementia and psychosis or agitation: secondary analysis of a double-blind, placebo-controlled trial. AB - OBJECTIVE: Authors evaluated the association between use/dosage of risperidone (RIS) and falls in a residential-care dementia population. METHODS: Authors performed secondary analysis of data from ambulatory patients in a randomized, double-blind, placebo-controlled, 12-week trial of three RIS dosages (0.5 mg/day, 1 mg/day, 2 mg/day). Outcomes included number of fallers, rate of falls, and time until the first fall after randomization. Additional analyses evaluated wandering as a potential moderating or mediating variable. RESULTS: The ambulatory sample included 537 subjects. Of those, 22.3% on placebo, 18.0% on RIS 0.5 mg/day, 12.7% on 1 mg/day, and 27.3% on 2 mg/day, respectively, fell during the trial. The difference between the RIS 1 mg/day group and placebo was significant, with a significantly lower hazard ratio in the RIS 1-mg/day group than placebo. Wandering was associated with an increased risk of falls. Among 205 patients with the highest levels of wandering at baseline, RIS 1 mg/day was associated with approximately a 70% reduction in risk for falls versus placebo condition. However, in those with the lowest levels of wandering at baseline, RIS 2 mg/day may have increased the risk of falls. CONCLUSIONS: Evaluating the benefits versus risks of risperidone in patients with dementia is complex and must consider multiple outcomes as a function of dose. At 1 mg/day, RIS was associated with decreased falls, especially in patients who exhibit wandering. However, at 2 mg/day, it may increase the risk of falls in ambulatory individuals with low levels of wandering. PMID- 15353389 TI - A randomized, double-blind, crossover comparison of risperidone and haloperidol in Korean dementia patients with behavioral disturbances. AB - OBJECTIVE: Behavioral disturbances in dementia are extremely prominent and distressful, and often result in serious physical, social, and economic consequences. The authors compared the efficacy and tolerability of risperidone and haloperidol in the treatment of behavioral and psychological symptoms of dementia (BPSD) in institutionalized elderly Korean patients with Alzheimer disease, vascular dementia, or mixed dementia. METHODS: This was an 18-week double-blind, crossover study involving 120 patients who were randomly assigned to receive flexible doses (0.5-1.5 mg/day) of risperidone or haloperidol. BPSD were assessed using the Korean version of the Behavioral Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD-K), the Korean version of the Cohen-Mansfield Agitation Inventory (CMAI-K), and the Clinical Global Impression of Change scale (CGI-C). Safety and tolerability assessments included the Extrapyramidal Symptom Rating Scale and the incidence of adverse events. RESULTS: Both risperidone and haloperidol were efficacious in alleviating BPSD. However, when receiving risperidone, patients showed significantly greater improvement than when receiving haloperidol in the total and subscale scores of the BEHAVE-AD-K, the total and subscale scores of the CMAI-K, and the scores on the CGI-C scale. Also, risperidone had an additional benefit on aggressiveness and anxieties/phobias. The risk of antipsychotic-induced parkinsonism throughout this study was significantly lower with risperidone than with haloperidol. CONCLUSION: Risperidone had a favorable efficacy and tolerability profile compared with haloperidol in the treatment of BPSD in this patient population. PMID- 15353390 TI - Evaluation of insomnia and daytime napping in Iranian Alzheimer disease patients: relationship with severity of dementia and comparison with normal adults. AB - OBJECTIVE: Alzheimer disease (AD) is among the most common and disabling diseases in adulthood, and it has been largely neglected in traditional societies like Iran. Exacerbation of AD symptoms creates many severe problems, especially in traditional Iranian extended families. One of these problems is insomnia, which can very significantly affect patients' family members because of disturbing insomnia-related behaviors. METHODS: The authors studied 53 patients with probable AD who had been interviewed for dementia and depression in the Neurology Clinic of Roozbeh Psychiatric Hospital (Tehran University of Medical Sciences). Excluding patients with overlapping multi-infarct dementia, 35 AD patients (22 men, 13 women) and 21 adults without dementia were studied as a comparison group (11 men, 10 women). RESULTS: There was a significant relationship between duration of informant-reported symptoms and severity of dementia with daytime napping and more daytime napping among AD patients than in the comparison group. There was also a significant relationship between cognitive decline and daytime napping in the comparison group. Our study showed a relationship between insomnia and aggression, between daytime napping and paranoid delusions; and between onset of insomnia and anxiety. The comparison group showed a significant relationship between depression and daytime napping. CONCLUSIONS: Daytime napping and insomnia are very common among Iranian AD patients, much more so than in normal adults. There is also a significant relationship among psychiatric symptoms, daytime napping, and insomnia, but it seems that daytime napping and insomnia in depressed AD patients are caused more by the progress of AD than by depression. PMID- 15353391 TI - Hemoglobin levels and Alzheimer disease: an epidemiologic study in India. AB - OBJECTIVE: Anemia is common in developing countries, where populations are aging rapidly. The authors explored the cross-sectional relationship between hemoglobin concentration and Alzheimer disease (AD) in a rural elderly sample in Ballabgarh, India. METHODS: A clinical diagnostic evaluation for dementia and a hemoglobin estimation were performed in 605 persons selected by screening a larger community based sample age 55+ years. Twenty-six participants met criteria for AD. RESULTS: Hemoglobin was inversely associated with AD after adjustment for age, sex, and literacy. CONCLUSION: Low hemoglobin is associated with AD and should be investigated further as a modifiable risk factor. PMID- 15353392 TI - Assessment of mood state in dementia by use of the visual analog mood scales (VAMS). AB - OBJECTIVE: The assessment of mood states in individuals with dementia is a challenging yet clinically useful task. The purpose of the present study was to examine the validity of the Visual Analog Mood Scales (VAMS) in individuals with dementia. METHODS: Thirty-one patients who met diagnostic criteria for dementia completed the VAMS and a modified Profile of Mood States. RESULTS: Authors found good convergent validity between all monotrait-heteromethod mood states. Excellent discriminant validity was found for VAMS Happy, Confused, Angry, and Energetic scales. CONCLUSION: These results provide evidence for the validity of the VAMS in patients with dementia. PMID- 15353393 TI - Late-life depression among black and white elderly homecare patients. AB - OBJECTIVE: The authors compared the prevalence of major depressive disorder (MDD) and the prescription rates of antidepressant medication, by race, among frail, older homecare patients. METHODS: A random sample of 56 black and 458 white newly admitted homecare patients age 65 and over was assessed for MDD with structured interviews and medical records, and antidepressant prescription rates were tallied. RESULTS: The prevalence of MDD did not differ significantly across racial groups. Only 16.7% of black patients and 32.0% of white patients were prescribed antidepressant medication. CONCLUSIONS: Prevalence of MDD was similar among black and white elderly homecare patients. In both groups, depression is undertreated and contributes to the burden of this frail, older patient group. PMID- 15353394 TI - Preliminary report on childhood sexual abuse, suicidal ideation, and suicide attempts among middle-aged and older depressed women. AB - OBJECTIVE: Authors examined the relationship between childhood sexual abuse histories and suicidal ideation and behavior among depressed women age 50 years and older. METHODS: After admission to a psychiatric unit, participants were administered the Structured Clinical Interview for DSM-III-R and measures of suicidal ideation and behavior. RESULTS: Women who reported abuse histories were more likely to report suicidal ideation at the time of hospitalization and a history of multiple suicide attempts. CONCLUSIONS: These preliminary findings underscore the need for more study of how childhood abuse amplifies risk for suicidal ideation and behavior among women across the life course. PMID- 15353395 TI - Characteristics and treatment outcome of older methadone-maintenance patients. AB - OBJECTIVE: There are over 160,000 persons receiving methadone maintenance treatment in the United States. Surveys indicate that 5% to 6% of these patients are older than age 55. Authors hypothesized, from a survey of methadone programs, that methadone patients older than age 55 would have higher rates of medical and psychiatric problems and higher rates of unemployment, and would do less well in treatment, as compared with their younger counterparts. METHODS: Authors reviewed 759 consecutive admissions to a Midwestern methadone-maintenance program. RESULTS: It was unexpectedly found that patients older than age 55 (N=54) were not different from their younger counterparts in terms of medical and psychiatric problems or employment; they were more likely to be married, and did significantly better in treatment. CONCLUSIONS: Older methadone patients may, in fact, have fewer problems and do well in treatment. This finding has implications regarding allocation of resources for methadone programs serving these patients. PMID- 15353396 TI - Donepezil in the treatment of dementia with lewy bodies. PMID- 15353397 TI - Diminished stress resilience in institutionalized elderly patients: is hypovitaminosis D a factor? PMID- 15353398 TI - Repeat cognitive screening of initially normal, older primary-care patients. PMID- 15353399 TI - The human paracellin-1 gene (hPCLN-1): renal epithelial cell-specific expression and regulation. AB - Tubular reabsorption of Mg2+ is mediated by the tight junction protein paracellin 1, which is encoded by the gene PCLN-1 (CLDN16) and exclusively expressed in the kidney. Tubular Mg2+ reclamation is modulated by many hormones and factors. The aim of this study was to define regulatory elements essential for renal tubular cell-specific expression of human PCLN-1 (hPCLN-1) and to explore the effect of Mg2+ transport modulators on the paracellin-1 gene promoter. Endogenous paracellin-1 mRNA and protein were detected in renal cell lines opossom kidney (OK), HEK293, and MDCT, but not in the fibroblast cell line NIH3T3. A 7.5-kb hPCLN-1 5'-flanking DNA sequence along with seven 5'-deletion products were cloned into luciferase reporter vectors and transiently transfected into the renal and nonrenal cells. The highest levels of luciferase activity resulted from transfection of a 5'-flanking 2.5-kb fragment (pJ2M). This activity was maximal in OK cells, was orientation dependent, and was absent in NIH3T3 cells. Mg2+ deprivation significantly increased pJ2M-driven activity in transfected OK cells, whereas Mg2+ load decreased it compared with conditions of normal Mg2+. Deletion analysis along with electrophoretic mobility-shift assay demonstrated that OK cells contain nuclear proteins, which bind a 70-bp region between -1633 and -1703 of major functional significance. Deleting this 70-bp segment, which contains a single peroxisome proliferator-response element (PPRE), or mutating the PPRE, caused a 60% reduction in luciferase activity. Stimulating the 70-bp sequence with 1,25(OH)2 vitamin D decreased luciferase activity by 52%. This effect of 1,25(OH)2 vitamin D was abolished in the absence of PPRE or in the presence of mutated PPRE. We conclude that the PPRE within this 70-bp DNA region may play a key role in the cell-specific and regulatory activity of the hPCLN-1 promoter. Ambient Mg2+ concentration and 1,25(OH)2 vitamin D may modulate paracellular, paracellin-1-mediated, Mg2+ transport at the transcriptional level. 1,25(OH)2 vitamin D exerts its activity on the hPCLN-1 promoter likely via the PPRE site. PMID- 15353400 TI - Stress response inhibits the nephrotoxicity of cisplatin. AB - Salt loading and saline hydration are used to protect patients from cisplatin induced nephrotoxicity. The mechanism by which salt exerts its protective effect is unknown. As part of an ongoing study of cisplatin nephrotoxicity, an in vitro assay system was developed that models the in vivo exposure and response of proximal tubule cells to cisplatin. In this study, it was discovered that the toxicity of cisplatin toward LLC-PK(1) cells varied dramatically according to the tissue culture media used for 3-h cisplatin exposure. Further experiments revealed that minor variations in the sodium concentration among standard tissue culture media modulated cisplatin nephrotoxicity. NaCl has been shown to protect against cisplatin-induced nephrotoxicity in vivo but has never before been demonstrated in vitro. NaCl did not alter the cellular accumulation of cisplatin. NaCl altered the osmolarity of the external media, and its effect was replicated by substituting equiosmolar concentrations of impermeant anions or cations. The change in osmolarity triggered a stress response within the cell that modulated sensitivity to cisplatin. These data resolve several long-standing controversies regarding the mechanism by which salt loading protects the kidney from cisplatin induced nephrotoxicity. PMID- 15353401 TI - Minocycline reduces renal microvascular leakage in a rat model of ischemic renal injury. AB - Tetracyclines exhibit significant anti-inflammatory properties, inhibit matrix metalloproteinases (MMPs), and are protective in models of ischemia-reperfusion injury (IRI). Both inflammatory cascades and MMP activation have been demonstrated to modulate microvascular permeability. Because increased microvascular permeability occurs during IRI in a variety of organ systems including the kidney, we hypothesized that minocycline, a semisynthetic tetracycline, would diminish microvascular leakage during renal IRI. To test this hypothesis, we used intravital 2-photon microscopy to examine leakage of fluorescent dextrans from the vasculature in a rodent model of IRI. Minocycline significantly reduced the extent of dextran (500 kDa) leakage from the renal microvasculature 24 h after ischemia. Although minocycline diminished leukocyte accumulation in the kidney following ischemia, areas of leukocyte accumulation did not correlate with areas of microvascular permeability in either the saline- or minocycline-pretreated animals. Minocycline diminished the perivascular increase in MMP-2 and MMP-9, as well as the increase in MMP-2 activity 24 h after ischemia. ABT-518, a specific inhibitor of MMP-2 and MMP-9, also significantly reduced the extent of dextran (500 kDa) leakage from the renal microvasculature 24 h after ischemia. Our results indicate that minocycline mitigates the renal microvascular permeability defect following IRI. This effect is spatially distinct from the effect of minocycline on leukocyte accumulation and may be related to diminished activity of MMPs on the integrity of the perivascular matrix. PMID- 15353402 TI - Expression of epidermal growth factor in the developing rat kidney. AB - Epidermal growth factor (EGF) is important in mammalian renal development. In our study, we investigated the detailed distribution and the time of the first appearance of EGF in developing rat kidney. Kidneys from embryonic 18 (E18)- and 20-day-old (E20) fetuses, postnatal 1 (P1)-, 3 (P3)-, 7 (P7)-, 14 (P14)-, and 21 day-old (P21) pups, and adults were processed for immunohistochemistry and electronmicroscopy. In adult rat kidney, EGF immunoreactivity was found in distal tubule including the thick ascending limb (TAL) and portion 1 of distal convoluted tubule (DCT1), whereas no EGF immunoreactivity was seen in portion 2 of distal convoluted tubule (DCT2) and connecting tubule. In developing kidney, EGF-positive cells first appeared at P3 and were localized in the middle portion of the differentiating TAL of the corticomedullary junction. By P7, the abundance of EGF expression had dramatically increased in the medullary TAL. Between P14 and P21, EGF immunoreactivity was found in the TAL and the DCT for the first time. However, EGF-positive and EGF-negative cells were in the TAL in developing rat kidney. EGF-positive cells did not differ from negative cells in the expression of sodium transport proteins or in the proliferation rate at P3 and P7. In the TAL, smooth-surfaced cells had strong EGF immunoreactivity, but no EGF immunoreactivity was seen in the rough-surfaced cells with well-developed microvilli. Our results suggest that the expression of EGF in developing kidney plays an important role in the regulation of growth and differentiation of the loop of Henle during kidney development and that this may act in the paracrine mode. PMID- 15353403 TI - A high-salt diet stimulates thick ascending limb eNOS expression by raising medullary osmolality and increasing release of endothelin-1. AB - A high-salt diet increases renal endothelin (ET) production and thick ascending limb (THAL) endothelial nitric oxide synthase (eNOS) expression. ET stimulates THAL eNOS expression via ET(B) receptors. The tonicity of the renal medulla is highly variable, and hyperosmolality stimulates ET-1 synthesis by endothelial cells. We hypothesized that a high-salt diet raises medullary osmolality, increases ET release by the THAL, and thus enhances eNOS expression. Seven days of high salt (1% NaCl in drinking water) increased eNOS expression in THALs by 125 +/- 31%. High salt increased outer medullary osmolality from 362 +/- 13 to 423 +/- 6 mosmol/kg H(2)O (P < 0.05). Bosentan, a dual-ET receptor antagonist, blocked the increase in THAL eNOS expression caused by high salt (2.66 +/- 0.44 absorbance units with bosentan vs. 5.15 +/- 0.67 for vehicle; P < 0.05). Conscious systolic blood pressure did not differ between the two groups. In primary cultures of medullary THALs, raising osmolality from 300 to 350 and 400 mosmol/kg H(2)O using NaCl increased eNOS expression by 39 +/- 11% (P < 0.05) and 71 +/- 16%, respectively (P < 0.05). In primary cultures of THALs, raising osmolality from 300 to 400 mosmol/kg H(2)O for 1 h increased ET-1 release from 62 +/- 7 to 113 +/- 2 pg/mg protein (P < 0.05). BQ-788, an ET(B) receptor antagonist (1 muM), blocked the stimulatory effect of 400 mosmol/kg H(2)O on eNOS expression (70 +/- 13% vs. -5 +/- 10%; paired difference, 74 +/- 15%; P < 0.05). BQ-788 alone had no significant effect. We concluded that high salt stimulates THAL eNOS expression by increasing outer medullary osmolality, ET-1 release by the THAL and ET(B) receptor activation. This may be an important regulatory mechanism of THAL NaCl absorption when dietary salt intake is increased. PMID- 15353404 TI - Cyclosporin A produces distal renal tubular acidosis by blocking peptidyl prolyl cis-trans isomerase activity of cyclophilin. AB - Cyclosporin A (CsA), a widely used immunosuppressant, causes distal renal tubular acidosis (dRTA). It exerts its immunosuppressive effect by a calcineurin inhibitory complex with its cytosolic receptor, cyclophilin A. However, CsA also inhibits the peptidyl prolyl cis-trans isomerase (PPIase) activity of cyclophilin A. We studied HCO(3)(-) transport and changes in beta-intercalated cell pH on luminal Cl(-) removal in isolated, perfused rabbit cortical collecting tubules (CCDs) before and after exposure to media pH 6.8 for 3 h. Acid incubation causes adaptive changes in beta-intercalated cells by extracellular deposition of hensin (J Clin Invest 109: 89, 2002). Here, CsA prevented this adaptation. The unidirectional HCO(3)(-) secretory flux, estimated as the difference between net flux and that after Cl(-) removal from the lumen, was -6.7 +/- 0.2 pmol.min( 1).mm(-1) and decreased to -1.3 +/- 0.2 after acid incubation. CsA in the bath prevented the adaptive decreases in HCO(3)(-) secretion and apical Cl(-):HCO(3)( ) exchange. To determine the mechanism, we incubated CCDs with FK-506, which inhibits calcineurin activity independently of the host cell cyclophilin. FK-506 did not prevent the acid-induced adaptive decrease in unidirectional HCO(3)(-) secretion. However, [AD-Ser](8) CsA, a CsA derivative, which does not inhibit calcineurin but inhibits PPIase activity of cyclophilin A, completely blocked the effect of acid incubation on apical Cl(-):HCO(3)(-) exchange. Acid incubation resulted in prominent "clumpy" staining of extracellular hensin and diminished apical surface of beta-intercalated cells [smaller peanut agglutinin (PNA) caps]. CsA and [AD-Ser](8) CsA prevented most hensin staining and the reduction of apical surface; PNA caps were more prominent. We suggest that hensin polymerization around adapting beta-intercalated cells requires the PPIase activity of cyclophilins. Thus CsA is able to prevent this adaptation by inhibition of a peptidyl prolyl cis-trans isomerase activity. Such inhibition may cause dRTA during acid loading. PMID- 15353405 TI - Characteristics of renal Rhbg as an NH4(+) transporter. AB - Rhbg is one of two recently cloned nonerythroid glycoproteins belonging to the Rh antigen family. Rhbg is expressed in basolateral membranes of intercalated cells of the kidney cortical collecting duct and some other cell types of the distal nephron and may function as NH(4)(+) transporters. The aim of this study was to characterize the role of Rhbg in transporting NH(4)(+). To do so, we expressed Rhbg in Xenopus laevis oocytes. Two-electrode voltage-clamp and H(+)-selective microlectrodes were used to measure NH(4)(+) currents, current-voltage plots, and intracellular pH (pH(i)). In oocytes expressing Rhbg, 5 mM NH(4)(+) induced an inward current of 93 +/- 7.7 nA (n = 20) that was significantly larger than that in control oocytes of -29 +/- 7.1 nA (P < 0.005). Whole cell conductance, at all tested potentials (-60 to +60 mV), was significantly more in oocytes expressing Rhbg compared with H(2)O-injected oocytes. In Rhbg oocytes, 5 mM NH(4)(+) depolarized the oocyte by 28 +/- 3.6 mV and decreased pH(i) by 0.30 +/- 0.04 at a rate of -20 +/- 2.5 x 10(-4) pH/s. In control oocytes, 5 mM NH(4)(+) depolarized V(m) by only 20 +/- 5.8 mV and pH(i) decreased by 0.07 +/- 0.01 at a rate of -2.7 +/- 0.6 x 10(-4) pH/s. Raising bath [NH(4)(+)] in increments from 1 to 20 mM elicited a proportionally larger decrease in pH(i) (DeltapH(i)), larger depolarization (DeltaV(m)), and a faster rate of pH(i) decrease. Bathing Rhbg oocytes in 20 mM NH(4)(+) induced an inward current of 140 +/- 7 nA that was not significantly different from 178 +/- 23 nA induced in H(2)O-injected (control) oocytes. The rate of pH(i) decrease induced by increasing external [NH(4)(+)] was significantly faster in Rhbg than in H(2)O-injected oocytes at all external NH(4)(+) concentrations. In oocytes expressing Rhbg, net NH(4)(+) influx (estimated from NH(4)(+)-induced H(+) influx) as a function of external [NH(4)(+)] saturated at higher [NH(4)(+)] with a V(max) of approximately 30.8 and an apparent K(m) of 2.3 mM (R(2) = 0.99). These data strongly suggest that Rhbg is a specific electrogenic transporter of NH(4)(+). PMID- 15353406 TI - Early changes in insulin secretion and action induced by high-fat diet are related to a decreased sympathetic tone. AB - To evaluate the relationship between the development of obesity, nervous system activity, and insulin secretion and action, we tested the effect of a 2-mo high fat diet in rats (HF rats) on glucose tolerance, glucose-induced insulin secretion (GIIS), and glucose turnover rate compared with chow-fed rats (C rats). Moreover, we measured pancreatic and hepatic norepinephrine (NE) turnover, as assessment of sympathetic tone, and performed hypothalamic microdialysis to quantify extracellular NE turnover. Baseline plasma triglyceride, free fatty acid, insulin, and glucose concentrations were similar in both groups. After 2 days of diet, GIIS was elevated more in HF than in C rats, whereas plasma glucose time course was similar. There was a significant increase in basal pancreatic NE level of HF rats, and a twofold decrease in the fractional turnover constant was observed, indicating a change in sympathetic tone. In ventromedian hypothalamus of HF rats, the decrease in NE extracellular concentration after a glucose challenge was lower compared with C rats, suggesting changes in overall activity. After 7 days, insulin hypersecretion persisted, and glucose intolerance appeared. Later (2 mo), there was no longer insulin hypersecretion, whereas glucose intolerance worsened. At all times, HF rats also displayed hepatic insulin resistance. On day 2 of HF diet, GIIS returned to normal after treatment with oxymetazoline, an alpha(2A)-adrenoreceptor agonist, thus suggesting the involvement of a low sympathetic tone in insulin hypersecretion in response to glucose in HF rats. In conclusion, the HF diet rapidly results in an increased GIIS, at least in part related to a decreased sympathetic tone, which can be the first step of a cascade of events leading to impaired glucose homeostasis. PMID- 15353407 TI - Effects of glucagon-like peptide-1 on endothelial function in type 2 diabetes patients with stable coronary artery disease. AB - GLP-1 stimulates insulin secretion, suppresses glucagon secretion, delays gastric emptying, and inhibits small bowel motility, all actions contributing to the anti diabetogenic peptide effect. Endothelial dysfunction is strongly associated with insulin resistance and type 2 diabetes mellitus and may cause the angiopathy typifying this debilitating disease. Therefore, interventions affecting both endothelial dysfunction and insulin resistance may prove useful in improving survival in type 2 diabetes patients. We investigated GLP-1's effect on endothelial function and insulin sensitivity (S(I)) in two groups: 1) 12 type 2 diabetes patients with stable coronary artery disease and 2) 10 healthy subjects with normal endothelial function and S(I). Subjects underwent infusion of recombinant GLP-1 or saline in a random crossover study. Endothelial function was measured by postischemic FMD of brachial artery, using ultrasonography. S(I) [in (10(-4) dl.kg(-1).min(-1))/(muU/ml)] was measured by hyperinsulinemic isoglycemic clamp technique. In type 2 diabetic subjects, GLP-1 infusion significantly increased relative changes in brachial artery diameter from baseline FMD(%) (3.1 +/- 0.6 vs. 6.6 +/- 1.0%, P < 0.05), with no significant effects on S(I) (4.5 +/- 0.8 vs. 5.2 +/- 0.9, P = NS). In healthy subjects, GLP-1 infusion affected neither FMD(%) (11.9 +/- 0.9 vs. 10.3 +/- 1.0%, P = NS) nor S(I) (14.8 +/- 1.8 vs. 11.6 +/- 2.0, P = NS). We conclude that GLP-1 improves endothelial dysfunction but not insulin resistance in type 2 diabetic patients with coronary heart disease. This beneficial vascular effect of GLP-1 adds yet another salutary property of the peptide useful in diabetes treatment. PMID- 15353408 TI - Mest/Peg1 imprinted gene enlarges adipocytes and is a marker of adipocyte size. AB - Obesity is a common and serious metabolic disorder in the developed world that is occasionally accompanied by type II diabetes, atherosclerosis, hypertension, and hyperlipidemia. We have found that mesoderm-specific transcript (Mest)/paternally expressed gene 1 (Peg1) gene expression was markedly enhanced in white adipose tissue of mice with diet-induced and genetically caused obesity/diabetes but not with streptozotocin-induced diabetes, which does not cause obesity. Administration of pioglitazone, a drug for type II diabetes and activator of peroxisome proliferator-activated receptor (PPAR)gamma, in obese db/db mice reduced the enhanced expression of Mest mRNA in adipose tissue, concomitant with an increase in body weight and a decrease in the size of adipose cells. Ectopic expression of Mest in 3T3-L1 cells caused increased gene expression of adipose markers such as PPARgamma, CCAAT/enhancer binding protein (C/EBP)alpha, and adipocyte fatty acid binding protein (aP)2. In transgenic mice overexpressing Mest in adipose tissue, enhanced expression of the adipose genes was observed. Moreover, adipocytes were markedly enlarged in the transgenic mice. Thus Mest appears to enlarge adipocytes and could be a novel marker of the size of adipocytes. PMID- 15353409 TI - Different epidemic curves for severe acute respiratory syndrome reveal similar impacts of control measures. AB - Severe acute respiratory syndrome (SARS) has been the first severe contagious disease to emerge in the 21st century. The available epidemic curves for SARS show marked differences between the affected regions with respect to the total number of cases and epidemic duration, even for those regions in which outbreaks started almost simultaneously and similar control measures were implemented at the same time. The authors developed a likelihood-based estimation procedure that infers the temporal pattern of effective reproduction numbers from an observed epidemic curve. Precise estimates for the effective reproduction numbers were obtained by applying this estimation procedure to available data for SARS outbreaks that occurred in Hong Kong, Vietnam, Singapore, and Canada in 2003. The effective reproduction numbers revealed that epidemics in the various affected regions were characterized by markedly similar disease transmission potentials and similar levels of effectiveness of control measures. In controlling SARS outbreaks, timely alerts have been essential: Delaying the institution of control measures by 1 week would have nearly tripled the epidemic size and would have increased the expected epidemic duration by 4 weeks. PMID- 15353410 TI - Invited commentary: real-time tracking of control measures for emerging infections. PMID- 15353412 TI - Sleep-disordered breathing, glucose intolerance, and insulin resistance: the Sleep Heart Health Study. AB - Clinic-based studies suggest that sleep-disordered breathing (SDB) is associated with glucose intolerance and insulin resistance. However, in the available studies, researchers have not rigorously controlled for confounding variables to assess the independent relation between SDB and impaired glucose metabolism. The objective of this study was to determine whether SDB was associated with glucose intolerance and insulin resistance among community-dwelling subjects (n=2,656) participating in the Sleep Heart Health Study (1994-1999). SDB was characterized with the respiratory disturbance index and measurements of oxygen saturation during sleep. Fasting and 2-hour glucose levels measured during an oral glucose tolerance test were used to assess glycemic status. Relative to subjects with a respiratory disturbance index of less than 5.0 events/hour (the reference category), subjects with mild SDB (5.0-14.9 events/hour) and moderate to severe SDB (> or =15 events/hour) had adjusted odds ratios of 1.27 (95% confidence interval: 0.98, 1.64) and 1.46 (95% confidence interval: 1.09, 1.97), respectively, for fasting glucose intolerance (p for trend < 0.01). Sleep-related hypoxemia was also associated with glucose intolerance independently of age, gender, body mass index, and waist circumference. The results of this study suggest that SDB is independently associated with glucose intolerance and insulin resistance and may lead to type 2 diabetes mellitus. PMID- 15353413 TI - Thirty-year trends in cardiovascular risk factor levels among US adults with diabetes: National Health and Nutrition Examination Surveys, 1971-2000. AB - Among US adults with diabetes, using data from the National Health and Nutrition Examination Survey for 1971-1974, 1976-1980, 1988-1994, and 1999-2000, the authors describe 30-year trends in total cholesterol, blood pressure, and smoking levels. Using Bayesian models, the authors calculated mean changes per year and 95% credible intervals for age-adjusted mean total cholesterol and blood pressure levels and the prevalence of high total cholesterol (> or =5.17 mmol/liter), high blood pressure (systolic blood pressure: > or =140 mmHg and/or diastolic blood pressure: > or =90 mmHg), and smoking. Between 1971-1974 and 1999-2000, mean total cholesterol declined from 5.95 mmol/liter to 5.48 mmol/liter (-0.02 (95% credible interval: -0.03, -0.01) mmol/liter per year). The proportion with high cholesterol decreased from 72% to 55%. Mean blood pressure declined from 146/86 mmHg to 134/72 mmHg (systolic blood pressure: -0.5 (95% credible interval: -1.1, 0.5) mmHg per year; diastolic blood pressure: -0.6 (95% credible interval: -1.0, 0.03) mmHg per year). The proportion with high blood pressure decreased from 64% to 37%, and smoking prevalence decreased from 32% to 17%. Although these trends are encouraging, still one of two people with diabetes has high cholesterol, one of three has high blood pressure, and one of six is a smoker. PMID- 15353414 TI - Glucose and insulin components of the metabolic syndrome are associated with hyperandrogenism in postmenopausal women: the atherosclerosis risk in communities study. AB - In 1990-1992, the authors investigated the association of total and free testosterone with the metabolic syndrome in postmenopausal US women not taking hormone replacement therapy (n=362) in a prevalent case-control study of carotid atherosclerosis. Free testosterone was estimated by using the free androgen index (FAI) (total testosterone/sex hormone-binding globulin ratio). The metabolic syndrome was defined as the presence of three or more of the following criteria: waist circumference > or =35 inches (88.9 cm), triglycerides > or =150 mg/dl, high density lipoprotein cholesterol <40 mg/dl, blood pressure >130/80 mmHg, fasting insulin > or =100 pmol/liter, or impaired glucose homeostasis (fasting glucose > or =110 mg/dl or diagnosed diabetes mellitus). FAI, but not total testosterone, was strongly associated with the metabolic syndrome. Compared with women in the lowest FAI quartile, those in the highest quartile had a fivefold greater odds of having the metabolic syndrome (odds ratio=5.38, 95% confidence interval: 2.70, 10.7) after adjustment for age, race, and carotid atherosclerosis status. In multivariate analyses, the three-component metabolic syndrome combinations that contained both hyperinsulinemia and hyperglycemia were most strongly associated with increased FAI (absolute increase=0.41-0.54 compared with that for women who did not have these combinations; all p's < 0.001). Higher FAI was associated with the hyperinsulinemia and hyperglycemia components of the metabolic syndrome. The role of androgens in glucose homeostasis in postmenopausal women requires further study. PMID- 15353415 TI - Low bone mineral density is related to echogenic carotid artery plaques: a population-based study. AB - In a 1994-1995 cross-sectional, population-based study of 2,543 men and 2,726 postmenopausal women aged 55-74 years in Tromso, Norway, the authors assessed a possible relation between bone mineral density (BMD) and the prevalence of carotid artery plaques, with an emphasis on plaque morphology. BMD measurements of the forearm and ultrasonography of the carotid artery were performed. Study participants were divided into quartiles with respect to sex-specific BMD values. Prevalent plaques were categorized into four groups ranging from low echogenicity to high echogenicity. For echogenic plaques, a significant inverse correlation with BMD was found (p for linear trend=0.007 after adjustment for age, sex, and cardiovascular risk factors). For predominantly echogenic plaques, a similar but weaker association was indicated (p = 0.08); for predominantly echolucent and echolucent plaques, no significant associations were observed (p > or = 0.3). Subjects whose BMD values were in the highest quartile had a statistically significant lower risk of echogenic plaques than subjects whose BMD values were in the lowest quartile (odds ratio=0.51, 95% confidence interval: 0.31, 0.83). This study indicates that low bone mass is associated with an increased risk of echogenic calcified atherosclerotic plaques but not with a risk of echolucent plaques. PMID- 15353416 TI - Impact of obesity on medical problems and quality of life in Taiwan. AB - Little is known about the impact of obesity on medical problems and quality of life for people in the Asia-Pacific region. This January 2002-June 2003 cross sectional study surveyed 6,318 Taiwanese (3,540 men and 2,778 women) visiting health screening centers in southern Taiwan. The authors used the body mass index classification endorsed by the World Health Organization for people in this region. Information was collected on 15 medical problems and quality of life outcomes, measured by the Medical Outcomes Study Short Form 36 questionnaire. After adjustment for age, lifestyle, and sociodemographic factors, and after comparison of subjects with those not overweight or obese (reference group), an increasing trend of body mass index effects based on this reference category was observed on hypertension, hypercholesterolemia, hypertriglyceridemia, type II diabetes, hyperuricemia, pulmonary function impairment, fatty liver disease, and osteoarthritis in both sexes (p <0.01). Concerning quality of life, an increasing trend of body mass index effects was also observed on the outcomes physical functioning and bodily pain for both sexes and role limitation due to physical problems for women (p <0.05). Specifically, only the physical functioning domain, including daily activities such as climbing stairs, bending, walking, or some moderate activities, was significantly associated with obesity and was limited to class II obesity. PMID- 15353417 TI - Antidepressant medication use and non-Hodgkin's lymphoma risk: no association. AB - Animal and human studies have suggested that antidepressant medications may be associated with several cancers. The authors evaluated the association between antidepressant medication use and the risk of non-Hodgkin's lymphoma using a Canadian population-based case-control study, the National Enhanced Cancer Surveillance Study. Non-Hodgkin's lymphoma cases (n=638) diagnosed in 1995-1996 were identified using the Ontario Cancer Registry, and controls (n=1,930) were identified from the Ontario Ministry of Finance Property Assessment Database. Antidepressant medication use was ascertained using a self-administered questionnaire. Multivariate logistic regression was used to estimate odds ratios. "Ever" use of antidepressant medications was not associated with non-Hodgkin's lymphoma risk. The odds ratio for non-Hodgkin's lymphoma with 25 or more months of tricyclic antidepressant medication use was 1.6; however, this was nonsignificant. Duration or history of use or individual types of antidepressant medications were not associated with non-Hodgkin's lymphoma risk. These findings do not support an increased risk of non-Hodgkin's lymphoma with antidepressant medication use. PMID- 15353418 TI - Intussusception and oral poliovirus vaccination: is there an association? AB - Oral rotavirus vaccine was voluntarily withdrawn from the market after studies observed an increased risk of intussusception within 2 weeks after immunization. Concern has been raised that other orally administered vaccines, such as oral poliovirus vaccine (OPV), may also be associated with intussusception. In this 1990-1998 case-control study, the authors examined the association between OPV and intussusception in the Washington State Medicaid population, evaluating receipt of OPV during the month prior to intussusception among 119 cases and 589 controls matched by date of birth. Analysis was conducted via matched conditional logistic regression, controlling for sex. Between 1990 and 1998, 119 children younger than age 2 years had a therapeutic enema, surgical reduction, or hospitalization for intussusception and had been enrolled in Medicaid for at least 1 month prior to their intussusception date. There was no significantly elevated risk of intussusception associated with receipt of OPV; 9.2% (11/119) of cases and 8.5% (50/589) of controls were given OPV 0-28 days prior to the case's intussusception date (odds ratio=1.1, 95% confidence interval: 0.5, 2.2). However, to address the hypothesis that risk of intussusception is related to receipt of a particular dose of OPV, a larger study would be required. PMID- 15353419 TI - Human immunodeficiency virus infection in a rural community in the United States. AB - In 1986, a population-based survey of human immunodeficiency virus (HIV) infection in a rural Florida community showed that HIV prevalence was 28/877 (3.2%, 95% confidence interval (CI): 2.0, 4.4). In 1998-2000, the authors performed a second population-based survey in this community and a case-control study to determine whether HIV prevalence and risk factors had changed. After 609 addresses had been randomly selected for the survey, 516 (85%) residents were enrolled, and 447 (73%) were tested for HIV. HIV prevalence was 7/447 (1.6%, 95% CI: 0.4, 2.7) in western Palm Beach County and 5/286 (1.7%, 95% CI: 0.2, 3.3) in Belle Glade (p=0.2 in comparison with 1986). Independent predictors of HIV infection in both 1986 and 1998-2000 were having a history of sexually transmitted disease, number of sex partners, and exchanging money or drugs for sex. A history of having sex with men was a risk factor among men in 1986 but not in 1998-2000; residence in specific neighborhoods was a risk factor in 1998-2000 but not in 1986. The authors conclude that heterosexually acquired HIV infection did not spread throughout the community between 1986 and 1998 but persisted at a low level in discrete neighborhoods. Interventions targeting HIV-endemic neighborhoods will be needed to further reduce HIV prevalence in this area. PMID- 15353420 TI - Added epidemiologic value to tuberculosis prevention and control of the investigation of clustered genotypes of Mycobacterium tuberculosis isolates. AB - The Centers for Disease Control and Prevention established the US National Tuberculosis Genotyping and Surveillance Network to study the utility of genotyping Mycobacterium tuberculosis isolates for prevention and control. From 1998 to 2000, four sites performed conventional contact investigations and epidemiologic investigations of cases with genotypically matched M. tuberculosis isolates, called cluster investigations. The authors compared cluster pairs (two cases with M. tuberculosis isolates having identical genotypes) whose epidemiologic linkages were discovered only during cluster investigation with those whose epidemiologic linkages were discovered during conventional contact investigation. Among the 2,141 reported culture-positive tuberculosis cases, 2,055 (96%) M. tuberculosis isolates were genotyped. By itself and at a minimum, cluster investigation added 43 (38%) of the 113 total epidemiologic linkages discovered. Of the epidemiologic linkages discovered during conventional contact investigation, 29% of tuberculosis case pairs were not supported by genotyping data. The linkages discovered only during cluster investigation were more likely discovered in nontraditional settings and relationships and among larger clusters (cluster size of >5: adjusted odds ratio = 57.6, 95% confidence interval: 31.8, 104.6). Information gained from genotyping M. tuberculosis isolates should initiate cluster investigations of tuberculosis cases not previously discovered as being epidemiologically linked during conventional contact investigation. Cluster investigations will play a crucial role in predicting recent tuberculosis transmission more accurately, as we move toward tuberculosis elimination in the United States. PMID- 15353421 TI - Comparability of skin screening histories obtained by telephone interviews and mailed questionnaires: a randomized crossover study. AB - The comparability of information collected through telephone interviews and information collected through mailed questionnaires has not been well studied. As part of the first phase of a randomized controlled trial of population screening for melanoma in Queensland, Australia, the authors compared histories of skin examination reported in telephone interviews and self-administered mailed questionnaires. A total of 1,270 subjects each completed a telephone interview and a mailed questionnaire 1 month apart in 1999; 564 subjects received the interview first, and 706 received the mailed questionnaire first. Agreement between the two methods was 91.2% and 88.6% for whole-body skin examination by a physician in the last 12 months and the last 3 years, respectively, and 81.9% for whole-body skin self-examination in the last 12 months. Agreement was lower for "any" skin self-examination. Agreement between the two methods was similar regardless of whether the interview or the questionnaire was administered first. Missing data were less frequent for interviews (0.5%) than for mailed questionnaires (3.8%). Costs were estimated at A$9.55 (US$6.21) per completed interview and A$3.01 (US$1.96) per questionnaire. The similarity of results obtained using telephone interviews and mailed questionnaires, coupled with the substantially higher cost of telephone interviews, suggests that self administered mailed questionnaires are an appropriate method of assessing this health behavior. PMID- 15353422 TI - Re: "condom effectiveness for reducing transmission of gonorrhea and chlamydia: the importance of assessing partner infection status". PMID- 15353423 TI - Re: "parity and the risk of Down's syndrome". PMID- 15353424 TI - Re: "parity and the risk of Down's syndrome". PMID- 15353425 TI - Cost-effectiveness of cardiac resynchronization therapy in patients with symptomatic heart failure. AB - BACKGROUND: Heart failure is a common, costly, and debilitating illness. Resynchronization of ventricular contraction in patients with heart failure improves ejection fraction. The long-term morbidity and costs associated with such cardiac resynchronization therapy remain unclear. OBJECTIVE: To assess the incremental cost-effectiveness of cardiac resynchronization therapy. DESIGN: Markov model with Monte Carlo simulation. Future costs and effects were discounted at 3%. DATA SOURCES: Effects data were obtained from a concurrent systematic review. Health-related quality-of-life and cost data were obtained from publicly available data or from surveys. TARGET POPULATION: Patients with reduced ventricular function and prolonged QRS. TIME HORIZON: Lifetime. PERSPECTIVE: U.S. health care system. INTERVENTIONS: Cardiac resynchronization therapy versus medical therapy. OUTCOME MEASURES: Quality-adjusted life-years (QALYs), costs, and incremental cost-effectiveness. RESULTS OF BASE-CASE ANALYSIS: Medical therapy yielded a median of 2.64 (interquartile range, 2.47 to 2.82) discounted QALYs and a median discounted lifetime cost of 34,400 dollars (interquartile range, 31,100 dollars to 37,700 dollars). Cardiac resynchronization therapy was associated with a median incremental cost of 107,800 dollars(interquartile range, 79,800 dollars to 156,500 dollars) per additional QALY. RESULTS OF SENSITIVITY ANALYSIS: Results were sensitive to changes in several variables, including the relative risk for death or hospitalization. LIMITATIONS: These results apply to patients who meet the inclusion criteria of the currently completed trials. CONCLUSIONS: The incremental cost per QALY for cardiac resynchronization is similar to that of other commonly used interventions but is sensitive to changes in several key variables. Resynchronization therapy should not be considered in patients with comorbid illness that shortens life expectancy. PMID- 15353426 TI - Location of adenomas missed by optical colonoscopy. AB - BACKGROUND: Previous estimates of the adenoma miss rate with optical colonoscopy (OC) are hindered by the use of OC as its own reference standard. OBJECTIVE: To evaluate the frequency and characteristics of colorectal neoplasms that are missed prospectively on OC by using virtual colonoscopy (VC) as a separate reference standard. DESIGN: Prospective, multicenter screening trial. SETTING: 3 medical centers. PARTICIPANTS: 1233 asymptomatic adults who underwent same-day VC and OC. MEASUREMENTS: Colorectal neoplasms (adenomatous polyps) missed at OC before VC results were unblinded. RESULTS: Fourteen (93.3%) of 15 nonrectal neoplasms were located on a fold; 10 (71.4%) of these were located on the backside of a fold. Five (83.3%) of 6 rectal lesions were located within 10 cm of the anal verge. LIMITATIONS: Estimation of the OC miss rate depended on polyp detection on both VC and second-look OC and therefore underestimates the true OC miss rate, particularly for smaller polyps. CONCLUSIONS: Most clinically significant adenomas missed prospectively on OC are located behind a fold or near the anal verge. The 12% OC miss rate for large adenomas (>or=10 mm) when state-of the-art 3-dimensional VC is used as a separate reference standard is increased from the previous 0% to 6% estimates derived by using OC as its own reference standard. PMID- 15353427 TI - Clinical characteristics and functional outcomes of West Nile Fever. AB - BACKGROUND: West Nile fever, considered a nonsevere manifestation of West Nile virus infection, has not been clinically well described in the United States. In 2002, Illinois had 884 documented cases of West Nile virus infection with 66 associated deaths. OBJECTIVE: To describe the symptoms and functional outcomes of West Nile fever. DESIGN: Case series. SETTING: Illinois. PATIENTS: 98 community dwelling patients with laboratory evidence of West Nile virus infection but no history of clinical evidence of meningitis, encephalitis, or acute flaccid paralysis. INTERVENTION: Outpatient interviews. MEASUREMENTS: Presence and duration of patient-reported symptoms of infection, symptom-associated absenteeism, health care use, and impact on daily activities. RESULTS: Of 98 patients, 96% had fatigue for a median of 36 days, 81% had fever for a median of 5 days, 71% had headache for a median of 10 days, 61% had muscle weakness for a median of 28 days, and 53% had difficulty concentrating for a median of 14 days. Thirty respondents reported hospitalization, with a median stay of 5 days. At 30 days after onset, 63% of respondents continued to have symptoms. Duration did not vary significantly with increased age. Among the 72 patients who normally attended work or school, 57 (79%) could not attend because of illness (median absence, 10 days). LIMITATIONS: Recall bias could have been introduced by the delay between illness onset and interview and by self-reporting of illness information. CONCLUSIONS: West Nile fever is a more severe illness than has previously been documented. Mandatory reporting of West Nile fever cases in addition to West Nile meningoencephalitis cases could allow more accurate and timely recognition of the geographic distribution of West Nile virus infections and could inform public health interventions. PMID- 15353428 TI - A controlled trial to increase detection and treatment of osteoporosis in older patients with a wrist fracture. AB - BACKGROUND: Despite the high risk for future fractures and the availability of effective treatments, fewer than 10% to 20% of patients who sustain a fragility fracture are tested or treated for osteoporosis. OBJECTIVES: To improve rates of testing and treatment for osteoporosis in patients with wrist fractures who are seen in the emergency department. DESIGN: Nonrandomized, controlled trial with blinded ascertainment of outcomes. SETTING: Emergency departments in Edmonton, Alberta, Canada. PATIENTS: Persons 50 years of age or older who were treated for a wrist fracture and their physicians. Patients admitted to the hospital or treated for osteoporosis were excluded. Overall, 572 consecutive patients with fractures were screened, and 102 patients (55 intervention, 47 control) and 101 physicians were studied. MEASUREMENTS: The primary end point was the prescription of osteoporosis treatment 6 months after fracture. Secondary end points included rates of testing for bone mineral density and patients' knowledge, satisfaction, and quality of life. INTERVENTION: Faxed physician reminders that contained osteoporosis treatment guidelines endorsed by local opinion leaders and patient education. Control patients received usual care and information about falls and home safety. RESULTS: The median patient age was 66 years. Most patients were female (78%) and white (79%); 70% of patients reported a previous fracture, and 22% had a fall with injury in the previous year. The intervention increased the rates of testing for bone mineral density to 62% (vs. 17% for controls; adjusted relative increase, 3.6 [P < 0.001]) and the rates of osteoporosis treatment to 40% (vs. 10% for controls; adjusted relative increase, 3.8 [P = 0.002]) within 6 months of fracture. Intervention patients were more likely to report a diagnosis of osteoporosis, but other patient-reported outcomes did not differ significantly between groups. LIMITATIONS: This was a small, nonrandomized, controlled study with process-based outcomes. CONCLUSIONS: In a multifaceted intervention directed at patients and their physicians, the rates of testing and treatment for osteoporosis after emergency department care for a fragility fracture were more than 3 times those of controls. PMID- 15353429 TI - Update in gastroenterology and hepatology. PMID- 15353430 TI - Systematic review: cardiac resynchronization in patients with symptomatic heart failure. AB - BACKGROUND: Even with optimal pharmacotherapy, symptomatic heart failure is associated with substantial morbidity and mortality. PURPOSE: To determine the efficacy and safety of cardiac resynchronization therapy in adults with advanced systolic heart failure. DATA SOURCES: The Cochrane Central Register of Controlled Trials (2002, volume 4), MEDLINE (1980-2003), EMBASE (1980-2003), other electronic databases, and U.S. Food and Drug Administration reports. We contacted primary study authors and device manufacturers, and we hand searched bibliographies of relevant papers and conference proceedings. STUDY SELECTION: Randomized, controlled clinical trials for efficacy and controlled trials plus prospective cohort studies for safety. DATA EXTRACTION: Two reviewers chose studies and extracted data independently; random-effects models were used for analyses. DATA SYNTHESIS: Nine trials were included in the efficacy review (3216 patients). All trial participants had reduced ejection fraction and prolonged QRS duration, and 85% had New York Heart Association (NYHA) class III or IV symptoms. Cardiac resynchronization therapy improved ejection fraction (weighted mean difference, 0.035 [95% CI, 0.015 to 0.055]), quality of life (weighted mean reduction in score on the Minnesota Living with Heart Failure Questionnaire, 7.6 points [CI, 3.8 to 11.5 points]), and function (58% vs. 37% of patients improved by at least 1 NYHA class). Heart failure hospitalizations were reduced by 32% (relative risk [RR], 0.68 [CI, 0.41 to 1.12]), with benefits most marked in patients with NYHA class III or IV symptoms at baseline (RR, 0.65 [CI, 0.48 to 0.88]; number needed to treat for benefit [NNT(B)], 12). All-cause mortality was reduced by 21% (RR, 0.79 [CI, 0.66 to 0.96]; NNT(B), 24), driven largely by reductions in death from progressive heart failure (RR, 0.60 [CI, 0.36 to 1.01]). Eighteen studies (total of 3701 patients with cardiac resynchronization devices) were included in the safety review. Implant success rate was 90% (CI, 89% to 91%), and 0.4% of patients died during implantation (CI, 0.2% to 0.7%). Over a median 6-month follow-up, leads dislodged in 9% of patients (CI, 7% to 10%) and mechanical malfunctions occurred in 7% (CI, 5% to 8%). LIMITATIONS: These trials enrolled only patients with heart failure with NYHA class III or IV symptoms despite medical therapy, a prolonged QRS duration, and reduced ejection fraction; in addition, experienced providers implanted the devices. Because all but one of these trials randomly assigned patients after device implantation, their results may overestimate the potential benefits of cardiac resynchronization. Finally, since few patients in these trials had bradyarrhythmias or atrial fibrillation, the role of cardiac resynchronization in such patients is uncertain. CONCLUSIONS: In selected patients with heart failure, cardiac resynchronization therapy improves functional and hemodynamic status, reduces heart failure hospitalizations, and reduces all-cause mortality. PMID- 15353431 TI - Assessing the new medicare prescription drug law. AB - The Medicare Modernization Act (MMA) is the product of a political compromise to attract moderate Republicans and enough Democrats without losing Republican conservatives. The compromise offered more private health plans to beneficiaries while maintaining and improving traditional Medicare's benefits. This compromise did not settle the debate over the legislation, which is a major issue in the 2004 elections. Voters poorly understand the law because of its complexity. In this paper, I explain how the policy decisions made by the U.S. Congress have contributed to the law's complexity and controversy. I examine the new private health plan options that will be offered to beneficiaries, improvements made to traditional Medicare, and the impact of introducing income-based determinations into Medicare. I also discuss the impact of the drug benefit on beneficiaries in different income and assets categories and Congress's decision to prohibit the federal government from directly negotiating prices with drug manufacturers. I conclude by assessing the major claims made by critics and proponents. Both might be more circumspect in their assessments of the law's impact, since it is impossible to predict how a law of such complexity, with so many human variables, will work out in the end. The MMA is a worthwhile but imperfect effort to extend drug coverage to seniors who are most in need. It deserves neither condemnation nor indiscriminate praise but instead a commitment to help it succeed. PMID- 15353432 TI - Colonoscopy: as good as gold? PMID- 15353433 TI - Physician as patient: lessons to learn. PMID- 15353434 TI - Malpractice reform and medical injury. PMID- 15353435 TI - New treatments for brittle bones. PMID- 15353436 TI - Aspirin use and risk for colorectal adenoma. PMID- 15353437 TI - Low-molecular-weight heparin for pulmonary embolism. PMID- 15353438 TI - Alcohol use and diabetes mellitus. PMID- 15353439 TI - Alcohol use and diabetes mellitus. PMID- 15353440 TI - Hospice effect on medicare expenditures. PMID- 15353441 TI - Silo-Filler's disease, the acute respiratory distress syndrome, and oxides of nitrogen. PMID- 15353442 TI - Summaries for patients. The cost-effectiveness of cardiac resynchronization therapy for heart failure. PMID- 15353443 TI - Summaries for patients. How often does colonoscopy fail to detect polyps? PMID- 15353444 TI - Summaries for patients. Clinical characteristics and consequences of West Nile Fever. PMID- 15353445 TI - Summaries for patients. Increasing the detection and treatment of osteoporosis in patients who present to an emergency department with a wrist fracture. PMID- 15353446 TI - Summaries for patients. Cardiac resynchronization therapy for heart failure. PMID- 15353447 TI - Noninvasive ventilation in exacerbations of chronic obstructive pulmonary disease: implications of different meta-analytic strategies. PMID- 15353448 TI - SNAP: workbench management tool for evolutionary population genetic analysis. AB - The reconstruction of population processes from DNA sequence variation requires the coordinated implementation of several coalescent-based methods, each bound by specific assumptions and limitations. In practice, the application of these coalescent-based methods for parameter estimation is difficult because they make strict assumptions that must be verified a priori and their parameter-rich nature makes the estimation of all model parameters very complex and computationally intensive. A further complication is their distribution as console applications that require the user to navigate through console menus or specify complex command-line arguments. To facilitate the implementation of these coalescent based tools we developed SNAP Workbench, a Java program that manages and coordinates a series of programs. The workbench enhances population parameter estimation by ensuring that the assumptions and program limitations of each method are met and by providing a step-by-step methodology for examining population processes that integrates both summary-statistic methods and coalescent-based population genetic models. AVAILABILITY: SNAP Workbench is freely available at http://snap.cifr.ncsu.edu. The workbench and tools can be downloaded for Mac, Windows and Unix operating systems. Each package includes installation instructions, program documentation and a sample dataset. SUPPLEMENTARY INFORMATION: A description of system requirements and installation instructions can be found at http://snap.cifr.ncsu.edu. PMID- 15353449 TI - Capturing biological information with class-responsibility-collaboration cards. AB - Class-responsibility-collaboration (CRC) cards have been used extensively in the software industry for defining complex object-oriented software requirements. We have adapted this tool to capture information about biological components, collaborators and responsibilities within these collaborations, which is not captured by current annotation tools. CRC cards should provide a common ground that will facilitate communication between biologist and computer scientists. AVAILABILITY: A CRC card template, XML representation and XML schema are freely available at http://people.musc.edu/~zhengw/CRCCard/CRC_Card_Index.html SUPPLEMENTARY INFORMATION: Supplemental Figures 1-4. PMID- 15353450 TI - iPfam: visualization of protein-protein interactions in PDB at domain and amino acid resolutions. AB - SUMMARY: There are many resources that contain information about binary interactions between proteins. However, protein interactions are defined by only a subset of residues in any protein. We have implemented a web resource that allows the investigation of protein interactions in the Protein Data Bank structures at the level of Pfam domains and amino acid residues. This detailed knowledge relies on the fact that there are a large number of multidomain proteins and protein complexes being deposited in the structure databases. The resource called iPfam is hosted within the Pfam UK website. Most resources focus on the interactions between proteins; iPfam includes these as well as interactions between domains in a single protein. AVAILABILITY: iPfam is available on the Web for browsing at http://www.sanger.ac.uk/Software/Pfam/iPfam/; the source-data for iPfam is freely available in relational tables via the ftp site ftp://ftp.sanger.ac.uk/pub/databases/Pfam/database_files/. PMID- 15353451 TI - A Bayesian approach to reconstructing genetic regulatory networks with hidden factors. AB - MOTIVATION: We have used state-space models (SSMs) to reverse engineer transcriptional networks from highly replicated gene expression profiling time series data obtained from a well-established model of T cell activation. SSMs are a class of dynamic Bayesian networks in which the observed measurements depend on some hidden state variables that evolve according to Markovian dynamics. These hidden variables can capture effects that cannot be directly measured in a gene expression profiling experiment, for example: genes that have not been included in the microarray, levels of regulatory proteins, the effects of mRNA and protein degradation, etc. RESULTS: We have approached the problem of inferring the model structure of these state-space models using both classical and Bayesian methods. In our previous work, a bootstrap procedure was used to derive classical confidence intervals for parameters representing 'gene-gene' interactions over time. In this article, variational approximations are used to perform the analogous model selection task in the Bayesian context. Certain interactions are present in both the classical and the Bayesian analyses of these regulatory networks. The resulting models place JunB and JunD at the centre of the mechanisms that control apoptosis and proliferation. These mechanisms are key for clonal expansion and for controlling the long term behavior (e.g. programmed cell death) of these cells. AVAILABILITY: Supplementary data is available at http://public.kgi.edu/wild/index.htm and Matlab source code for variational Bayesian learning of SSMs is available at http://www.cse.ebuffalo.edu/faculty/mbeal/software.html. PMID- 15353452 TI - Quality assessment in cardiac surgery: do not miss the boat! PMID- 15353453 TI - Slide tracheoplasty for congenital tracheal stenosis with glottic stenosis. AB - A 7-month-old child with respiratory distress was diagnosed to have full-length tracheal stenosis with glottic stenosis. Slide tracheoplasty and anterior cricoid split with stenting for glottic stenosis were successfully performed, and the child has completed 5 years of asymptomatic follow-up. This case widens the scope for slide tracheoplasty, and cricoid split with stenting is a feasible procedure for associated glottic stenosis. PMID- 15353454 TI - Capillary leakage in cardiac surgery with cardiopulmonary bypass. AB - Cardiopulmonary bypass causes a systemic inflammatory response, which can lead to capillary leak syndrome. In 15 adults undergoing elective cardiac surgery with cardiopulmonary bypass, we determined the volume and peak time of capillary leakage from the measurements of extracellular fluid volume and circulating blood volume taken preoperatively, at various intervals up to 24 hours after surgery, and on the 7th postoperative day. Extracellular fluid volume rose from 15.5 +/- 2.7 L preoperatively to a peak 4 hours after surgery of 18.3 +/- 3.2 L and remained elevated at 24 hours. Circulating blood volume fell from 4.10 +/- 0.68 L preoperatively to 3.20 +/- 0.58 L at the end of surgery. Fluid administered intraoperatively did not raise the circulating blood volume. Intraoperative fluid balance was positive at 2.62 +/- 0.72 L but negative at all time points postoperatively. There was significant postoperative capillary leakage, increasing from 4.7% +/- 2.3% of body weight at the end of surgery to a peak 4 hours later of 5.4% +/- 2.0% and falling to 2.8% +/- 3.3% at 24 hours. This knowledge of the pattern of change in capillary leakage after cardiac surgery with cardiopulmonary bypass might serve as a valuable guide for postoperative management. PMID- 15353455 TI - Blood-air interface during cardiopulmonary bypass. AB - The aim of this study was to compare the systemic blood activation with open and closed perfusion management during cardiopulmonary bypass. In 30 patients undergoing coronary artery bypass grafting, we prospectively studied systemic blood activation, blood loss and the need for donor blood. In 15 patients we used an open venous reservoir consisting of a hard shell venous reservoir with an integrated cardiotomy filter. In another 15 patients we used a totally closed venous reservoir consisting of a collapsible venous reservoir, no coronary suction, modified vent and cell saver. Venous blood samples were collected pre, post and 24 hours postoperatively. Sex, age and perfusion times were identical in both groups. There were no statistically significant differences in concentrations of FXIIa and C3a, amount of blood loss and need for donor blood. Interleukin-6 and Elastase levels showed trends toward a lesser inflammatory reaction in closed venous reservoir patients. Modification of perfusion management with optimized air management does not seem to be an effective strategy in reducing the inflammatory response and influencing the coagulation system in this small cohort. PMID- 15353456 TI - Factors affecting early exsanguination and death in blunt thoracic aortic trauma. AB - The traditional approach to blunt thoracic aortic injuries has been expedient diagnosis and operative repair due to the significant risk of early exsanguination and death in initial survivors. Nonoperative management has been advocated in patients with multiple injuries to reduce the operative mortality. However, specific clinical parameters and diagnostic tests that may predict the risk of early exsanguination and death have yet to be identified. A retrospective analysis of 80 patients with these injuries was undertaken to identify factors associated with early exsanguination or death. Available aortograms were also examined and graded to determine their utility in predicting these outcomes. Early exsanguination and death were found to be associated with low systolic blood pressure on admission and with short duration from injury to diagnosis. Exsanguination was also associated with the total number of lesions in thoracic injuries, and mortality with age greater than 30 years. Aortographic appearance was not found to correlate with either outcome. Patients with blunt thoracic aortic injuries should continue to be managed expediently, with immediate surgical repair if not contraindicated by associated injuries, to avoid early rupture. PMID- 15353457 TI - Long term outcome of femoropopliteal bypass for claudication and critical ischemia. AB - We performed 167 femoropopliteal bypass surgeries in 151 patients (95 patients underwent above-knee bypass and 56 below-knee bypass) from December 1985 to December 2000 with the use of prosthetic graft or autologous vein graft. We compared primary patency rates between age, sex, graft material, distal anastomotic site and severity of ischemia, considering their survival rates to elucidate the long-term outcome of above-knee and below-knee femoropopliteal bypass. The 10 year patency rate for above-knee bypass was 47.4%, compared to 36.9% for below-knee ( p < 0.01). Better results were found after bypass surgery for claudicants than for critical ischemia ( p < 0.05). With regard to graft material and age categories, there were unexpectedly no statistical differences in either above-knee or below-knee anastomosis. The survival rate at 10 years in claudicants was 51.2%, compared to 15.9% with critical ischemia ( p < 0.01). Mortality was much influenced by ischemic heart disease ( p < 0.002) and the age of patient ( p < 0.05). The results after above-knee bypass had comparable patency, whereas the results after below-knee bypass were disappointing. Below knee arterial reconstruction for claudicants should be carefully considered and might be recommended only to patients with critical ischemia. PMID- 15353458 TI - Aortic valve replacement with the medtronic freestyle stentless bioprosthesis. AB - During a five-year period from 1996 to 2000, the Medtronic Freestyle stentless bioprosthesis was implanted in 310 patients of advanced age. Age at operation ranged from 60 to 90 years (mean, 76 +/- 4 years). 191 patients were female and 119 male. All implants were done by the modified subcoronary method using our own modification which enabled an improved adaptation of the porcine aortic root to the human anatomy. Two sinuses were scalloped and the third left intact. Additional coronary bypass grafts were necessary in 129 (39%) patients and mitral valve procedures in 23 (7%). Mean perfusion time was 109 +/- 12 minutes and crossclamp time 87 +/- 8 minutes. 16 (5%) patients died perioperatively. Another 17 (5.7%) patients died during a 1 to 5.6 year follow-up (mean, 2.9 years). There was only one valve related death due to infection of the valve. In spite of the advanced age, 95% of the survivors were free from cardiac symptoms and continued to live an active and fruitful life. The biological nature of the valve and the low gradients are perhaps reasons for the good results. The long-term results are expected to be good. PMID- 15353459 TI - Coronary bypass surgery in patients aged 70 years and over: mortality, morbidity, length of stay and hospital cost. AB - The objective of this investigation was to compare the outcome of isolated coronary artery bypass grafting surgery in patients > or = 70 years with those < 70. The cardiac surgery database of the Institute was used to obtain the characteristics of patients undergoing coronary artery bypass grafting between January 2000 and September 2001. The patients were divided into those > or = 70 years of age and those < 70 years old. A Parsonnet risk score was determined for each patient for the analysis of mortality, length of stay and hospital charges. During the study period, 1594 cases of isolated coronary artery bypass grafting were carried out. 184 (11.5%) cases were performed in the older group. The 30-day mortality for patients aged 70 and over was 7 (3.3%) out of 184 while that of patients < 70 years of age was 47 (3.8%) out of 1410 (p < 0.740). The overall hospital mortality was 10 (5.4%) of 184 and 53 (3.8%) of 1410 (p < 0.272) respectively. Apart from a higher incidence of wound infection in elderly patients, the frequency of other major complications was comparable in both groups. The average length of postoperative stay for the elderly patients was 10.4 +/- 0.9 days compared to 8.7 +/- 0.2 days for the younger group (p < 0.049). The mean hospital charge in patients > or = 70 was RM 25,160.38 +/- 1656.75 whereas for patients < 70, it was RM 21,801.47 +/- 308.91 (p < 0.048). This study supports the continued performance of coronary artery bypass grafting in patients > or = 70 years. Advanced age alone should not deter a cardiac surgeon from offering such a potentially beneficial intervention. PMID- 15353460 TI - A new technique of fixing a costal coaptation pin after resection of rib segment. AB - The Poly-L-lactide costal coaptation pin is an effective device in chest wall reconstruction. However, fixation is sometimes incomplete, despite the use of the costal coaptation pin. We report here the use of two suture techniques for the fixation of the incised ribs with costal coaptation and discuss the effectiveness of these procedures. We used the Poly-L-lactide costal coaptation pin in 174 cases of posterolateral thoracotomies with two suture methods. In one method the rib was generally fixed with suture only (L-method, n = 30), and in the H-method pairs of holes were made at the end of the incised ribs for ligating with sutures (H-method, n = 144). The effectiveness of each method was evaluated based on the degree of fixation and lateral shift 24 months postoperatively. Lateral shift was none in 114 (79.2%) cases using the H-method and 18 (60.0%) cases using the L method. Fixation was good in 131 (91.0%) cases using the H-method but in only 20 (66.7%) cases using the L-method. H-method was significantly more effective than the L-method of costal coaptation. The H-method was very effective for fixing incised ribs and is convenient for use by thoracic surgeons. PMID- 15353461 TI - Semi-skeletonized internal mammary artery grafts and sternal wound complications. AB - This study aimed to evaluate the risk factors for sternal wound complications in patients undergoing myocardial revascularization using bilateral semi skeletonized internal mammary arteries. Prospectively collected data on 751 patients undergoing coronary artery surgery from September 1994 to August 2002 were analyzed. The mean age of the patients was 56 years, 633 (84%) were male, 44 (6%) were over 66 years of age, and 170 (23%) were diabetic. Forty-four (5.9%) patients developed sternal wound complications. Among these cases, sternal infection occurred in 22 (2.9%) patients, of which 15 (2.0%) had sternal infection with mediastinitis and 7 (0.9%) had sternal infection alone. Independent risk factors for any sternal wound complications were peripheral vascular disease, diet-controlled diabetes, and delayed sternal closure. The risk factors for sternal infection were diabetes, postoperative pulmonary complications, and postoperative stroke. The perioperative mortality rate was 1.5% (11 patients), including 2 patients who had sternal wound complications. The use of bilateral semi-skeletonized internal mammary artery conduits carries a comparable sternal wound complication rate as conduits harvested by other techniques. PMID- 15353462 TI - Transventricular mitral commissurotomy in critical mitral stenosis during pregnancy. AB - The management of a pregnant patient with mitral stenosis is a subject of debate with regards to the optimal type of treatment and the time of intervention. We performed trans-ventricular mitral commissurotomy (TVMC) either as an isolated procedure in the second trimester, or in combination with Cesarian section at term. We retrospectively analyzed our experience with TVMC during pregnancy and formulated a protocol for its management. Between January 1987 and April 2002, fifty one patients underwent TVMC for critical mitral stenosis during pregnancy. In 38 patients, elective TVMC was performed during the second trimester, while in 12, it was performed as an initial procedure along with Cesarian section at term. One patient had an emergency TVMC in the second trimester when she presented with intractable acute pulmonary edema. There were no maternal mortalities. Three patients who developed post-operative mitral regurgitation were managed conservatively. Another two patients who developed cerebral embolism with hemiplegia recovered completely without any neurological deficit. There was only one fetal death in a patient where TVMC was performed as an emergency procedure for acute pulmonary edema. We conclude that TVMC in pregnancy is a safe, cost effective alternative in critical mitral stenosis complicating pregnancy. PMID- 15353463 TI - Early results and problems with St. Jude Medical Symmetry bypass system in Japan. AB - The early results of coronary artery grafting with an aortic connector system were assessed in Japanese patients. From May 2002 through April 2003, 24 consecutive patients underwent off-pump coronary artery bypass using an aortic connector system. Another patient was excluded because the saphenous vein was insufficient for the smallest available aortic connector system. Saphenous veins were harvested from the thigh in 17 (70.8%) patients, and from the lower leg in 7. The size of the aortic connector system was 4.5-5.0 mm in 19 (79.2%) patients. Intraoperative epiaortic echo indicated that a side-clamp was contraindicated in 15 cases. Hemostasis was instantaneous in all patients. There were no hospital deaths and no neurologic morbidity. Pre-discharge angiography revealed 100% patency of the anastomoses. Use of the aortic connector system demonstrated excellent early results with low neurologic morbidity even when employed in the context of an atheromatous ascending aorta. However, smaller sizes of the device are required for some Japanese patients. PMID- 15353464 TI - Evolution of repaired and non-repaired tricuspid regurgitation in rheumatic mitral valve surgery without severe pulmonary hypertension. AB - The aim of this study was to evaluate the usefulness of repairing significant tricuspid regurgitation (> or = grade 2) without severe pulmonary hypertension (< or = 50 mm Hg). Between 1993 and June 2001, 88 consecutive patients were operated on for rheumatic mitral valve disease associated with significant tricuspid regurgitation and without severe pulmonary hypertension. The severity of the tricuspid valve disease was assessed by echocardiography. Sixty-three patients had severe (> or = grade 3) tricuspid regurgitation (Group I), and 25 patients had moderate (grade 2) tricuspid regurgitation (Group II). There was no hospital mortality. six patients died during follow-up. The overall actuarial survival rate for 8 years was 92.1% +/- 3.1%. Cox proportional hazard regression analysis showed that age ( p = 0.006) and pulmonary complication ( p = 0.01) were associated with increased late mortality. Freedom from death was similar in both groups at 8 years (93.1% +/- 3.3% versus 88% +/- 8%, p = 0.7). Severe postoperative tricuspid regurgitation (> or = grade 3), caused by the failure of tricuspid repair or leaving the valve untouched, impaired long-term survival after surgery, and actuarial survival was 96.1% +/- 2.7% and 83% +/- 7.8% at 7 years ( p = 0.048), respectively. Severe tricuspid regurgitation, functional or organic, should be corrected at the time of mitral valve surgery, whereas untouched functional moderate tricuspid regurgitation improves after mitral valve surgery. PMID- 15353465 TI - Surgical management of pulmonary cavity associated with fungus ball. AB - Surgery for pulmonary cavity associated with fungus ball is challenged by chronic lung disease. The purpose of this report was to review patient data, operative procedures and results of surgery. This was a retrospective study. Twenty patients were operated on between January 1997 and December 2002. Fourteen (70%) patients were male and the mean age was 46.30 +/- 13.10 years (range, 24 to 76 years). The most common underlying pulmonary disorder was tuberculosis (70%). Ninety five percent of the patients had a history of hemoptysis, and 35% presented with massive hemoptysis. Lobectomy was performed in 11 (55%) patients and 6 (30%) patients were operated on by cavernostomy with transposition of muscle flap technique. There was no operative mortality and 8 complications (3 prolonged airleaks, 2 wound infections, 1 postoperative bleeding, 1 seroma and 1 empyema). It was also found that emergency surgery and cavernostomy with transposition of muscle flap compromised the postoperative course. Surgery is very effective in controlling and preventing hemoptysis in patients who have pulmonary cavity associated with fungus ball. Elective surgery and formal pulmonary resection may be the proper option for low risk patients. Cavernostomy with transposition of muscle flap may be suitable for patients who have poor pulmonary reserve. PMID- 15353466 TI - What influences the results in critical patients after cardiovascular surgery? AB - The predictive factors of surgical outcome were evaluated in compromised patients following cardiovascular surgery. Of 608 patients undergoing cardiovascular surgery between 1991 and 1999, 55 stayed in the intensive care unit for 2 weeks or longer. The mean age of these 55 patients was 56 years. There were 35 survivors and 20 nonsurvivors. Postoperative respiratory failure and gastrointestinal complications were significantly more frequent in those who died. The survival rate was significantly higher in patients who had enteral feeding compared to those who did not (88% versus 43%). Serum cholinesterase and total cholesterol concentrations were higher in the survivors. It was concluded that postoperative respiratory and gastrointestinal conditions influenced the surgical outcome, and serum cholinesterase and total cholesterol concentrations were valuable predictors of survival. PMID- 15353467 TI - Ruptured chronic traumatic mycotic pseudoaneurysm of the ascending aorta. AB - We report a rare case of posttraumatic chronic mycotic pseudoaneurysm of the ascending aorta presenting with acute rupture. The uniqueness of the case lies in its unusual etiology, presentation, and management with direct repair of the aorta without using cardiopulmonary bypass. PMID- 15353468 TI - Atypical pulmonary giant hydatid cyst as bilaterally symmetrical solitary cysts. AB - A pulmonary giant hydatid cyst, a special clinical entity, is rare. Our case involves a young patient who presented with a bilaterally symmetrical solitary cyst in each lung, a feature consistent with congenital lung cysts. The radiological and immunological findings were equivocal. A diagnosis of giant hydatid cyst was made intraoperatively and both cysts were removed conservatively. A follow-up showed complete recovery. PMID- 15353469 TI - Combined coronary artery bypass grafting and lung surgery. AB - Combined coronary bypass and lung surgery was performed in 3 patients. Through a median sternotomy or a left thoracotomy, bypass grafting was performed on beating heart or under cardiopulmonary bypass, followed by the lung operation. The lung lesion was diagnosed as carcinoma in 2 patients and hydatid cyst in 1 patient. With few exceptions, beating heart coronary bypass through a median sternotomy can be performed in a combined operation. PMID- 15353470 TI - Posterior mediastinal goiter. AB - Most mediastinal goiters are retrosternally situated in the anterior mediastinal compartment. Posterior mediastinal goiters, either retrotracheal or retroesophageal, are rare. We herein describe a case involving a retrotracheal goiter in the right posterior mediastinum, which was excised using a combined cervico-partial sternotomy and right thoracotomy approach. PMID- 15353471 TI - Cusp extension technique for bicuspid aortic valve in Turner-like stigmata. AB - A 19-year-old girl with bicuspid aortic valve and Turner-like stigmata underwent a successful repair with a cusp extension technique using fresh autologous pericardium for aortic insufficiency. She also had a simultaneous artificial graft replacement for ascending aortic dilatation. Aortic valve function showed no significant stenosis and slight insufficiency 2 years postoperatively. This technique is recommended to avoid a Ross or Ross/Konno procedure because of its ready availability, simplicity and excellent midterm results. PMID- 15353472 TI - Sutureless fixation of temporary pacing wire. AB - A simple sutureless technique of fixing ordinary straight insulated temporary pacing wire is described by creating a floret of the insulation of the wire. It can be easily accomplished and reproduced. PMID- 15353473 TI - Radiofrequency ablation for atrial fibrillation: different approaches. AB - The existing literature regarding radiofrequency ablation for the surgical treatment of atrial fibrillation was reviewed, analyzing the early and late results. A MEDLINE search supplemented with a manual bibliographic review was performed for all peer-reviewed English language articles regarding the use of radiofrequency ablation for the treatment of atrial fibrillation. Six studies were identified, with a total of 451 patients. None of the studies was completely randomized. All patients underwent radiofrequency ablation as an adjunct to a variety of cardiac surgical procedures. The hospital mortality rate was 2.7%. The overall survival rate was 97.1%, and freedom from atrial fibrillation was 76.3% +/- 5.1% after a mean follow-up period of 13.8 +/- 1.9 months. It was concluded that radiofrequency ablation should be considered a safe and effective means to cure atrial fibrillation in patients undergoing open heart surgery. PMID- 15353474 TI - Asymptomatic cardiac lipoma as a finding in a left thoracotomy for lung cancer. PMID- 15353475 TI - Tracheobronchopathia osteochondroplastica as an incidental finding. PMID- 15353476 TI - Gender difference outcomes after coronary artery surgery. PMID- 15353477 TI - Abnormal regulation of Mg2+ transport via Na/Mg exchanger in sickle erythrocytes. AB - Erythrocyte magnesium (Mg2+) deficiency has been demonstrated in sickle cell disease to contribute to erythrocyte dehydration, K loss, and thus sickling. No studies have assessed the functional properties of the Na/Mg exchanger in sickle cell disease. Using Mg(2+)-loaded erythrocytes, we measured Mg2+ efflux induced by extracellular Na+. We estimated that the Na/Mg exchanger had higher maximal velocity, higher affinity for Na+, and lower cooperativity for Mg2+ in sickle than in normal erythrocytes. The activity of the exchanger was markedly decreased by hypotonic and hypertonic conditions in normal erythrocytes but not in sickle erythrocytes. Studies of density-separated erythrocytes showed that the activity of the exchanger decreased as the mean cellular hemoglobin concentration increased in normal but not in sickle erythrocytes. Inhibition of protein kinase C (PKC) activity by calphostin C and chelerythrine increased the activity of the exchanger in normal but not in sickle erythrocytes. Inhibition of serine/threonine phosphatases did not affect the activity of the exchanger in either normal or sickle erythrocytes. Altogether, these data indicate that the Na/Mg exchanger is abnormally regulated in sickle erythrocytes. Therefore, Mg2+ depletion in sickle erythrocytes might be mediated by an up-regulated Na/Mg exchanger, possibly by dephosphorylation of the transporter or a closely associated regulator. PMID- 15353478 TI - Modulation of cord blood CD8+ T-cell effector differentiation by TGF-beta1 and 4 1BB costimulation. AB - Transforming growth factor-beta1 (TGF-beta1), an immunosuppressive cytokine, inhibits cytotoxic T cell (CTL) immune responses. In contrast, 4-1BB (CD137), a costimulatory molecule in the tumor necrosis factor (TNF) receptor family, amplifies CTL-mediated antitumor immune responses. We investigated whether TGF beta1 responses could be reversed by 4-1BB costimulation during in vitro differentiation of naive CD8+ T cells into effector CTL cells. TGF-beta1 potently suppressed CTL differentiation of human cord blood naive CD8+ T cells as determined by reduced induction of characteristic phenotypes of effector cells and cytotoxic activity. TGF-beta1-mediated suppression of CTL differentiation was abrogated by 4-1BB costimulation but not by CD28 or another member in the TNF receptor family, CD30. 4-1BB costimulation suppressed Smad2 phosphorylation induced by TGF-beta1, suggesting that 4-1BB effects were at the level of TGF beta1 signaling. 4-1BB effects on the TGF-beta1-mediated suppression were enhanced by interleukin 12 (IL-12) but counteracted by IL-4; 4-1BB expression was up- or down-regulated, respectively, by IL-12 and IL-4. IL-4 was more dominant than IL-12 when both cytokines were present during 4-1BB costimulation in the presence of TGF-beta1. This indicates critical roles for IL-4 and IL-12 in regulating 4-1BB effects on TGF-beta1-mediated suppression. PMID- 15353479 TI - Dendritic cell-derived IL-2 production is regulated by IL-15 in humans and in mice. AB - Dendritic cells (DCs) are involved in the initiation and regulation of innate and adaptive immune responses. Several molecular mechanisms regulate these diverse DC functions, and we have previously reported that mouse dendritic cells (mDCs) can produce interleukin-2 (IL-2) in vitro and in vivo, in response to microbial activation and T-cell-mediated stimuli. This property is shared by different DC subtypes, including Langerhans cells. Here we show that, on appropriate stimulation, human DCs, both plasmacytoid and myeloid subtypes, also express IL 2. Interestingly, the production of IL-2 by myeloid DCs is induced by T-cell mediated stimuli and depends on the presence of IL-15. The key role of this cytokine in regulating IL-2 production was also confirmed in the mouse system. In particular, we could show that DCs from IL-15-deficient mice were strongly impaired in the ability to produce IL-2 after interactions with different microbial stimuli. Our results indicate that DC-produced IL-2 is tightly coregulated with the expression of IL-15. PMID- 15353480 TI - The CD28 family: a T-cell rheostat for therapeutic control of T-cell activation. AB - The CD28 family of receptors (CD28, cytotoxic T-lymphocyte-associated antigen 4 [CTLA-4], inducible costimulator [ICOS], program death-1 [PD-1], and B- and T lymphocyte attenuator [BTLA]) plays a critical role in controlling the adaptive arm of the immune response. While considerable information is available regarding CD28 and CTLA-4, the function of the more recently discovered members of the CD28 family is less well understood. This review will highlight recent findings regarding the CD28 family with special emphasis on effects the CD28 family has on immunopathology, the discovery of costimulatory antibodies with superagonist function, and the status of clinical trials using various strategies to augment or block T-cell costimulation. PMID- 15353481 TI - Hematopoietic stem cell transplantation (HSCT) in children with juvenile myelomonocytic leukemia (JMML): results of the EWOG-MDS/EBMT trial. AB - Allogeneic hematopoietic stem cell transplantation (HSCT) is the only proven curative therapy for juvenile myelomonocytic leukemia (JMML). We, the European Working Group on Childhood MDS (EWOG-MDS) and the European Blood and Marrow Transplantation (EBMT) Group, report the outcome of 100 children (67 boys and 33 girls) with JMML given unmanipulated HSCT after a preparative regimen including busulfan, cyclophosphamide, and melphalan. Forty-eight and 52 children received transplants from an HLA-identical relative or an unrelated donor (UD), respectively. The source of hematopoietic stem cells was bone marrow, peripheral blood, and cord blood in 79, 14, and 7 children, respectively. Splenectomy had been performed before HSCT in 24 children. The 5-year cumulative incidence of transplantation-related mortality and leukemia recurrence was 13% and 35%, respectively. Age older than 4 years predicted an increased risk of disease recurrence. The 5-year probability of event-free survival for children given HSCT from either a relative or a UD was 55% and 49%, respectively (P = NS), with median observation time of patients alive being 40 months (range, 6 to 144). In multivariate analysis, age older than 4 years and female sex predicted poorer outcome. Results of this study compare favorably with previously published reports. Disease recurrence remains the major cause of treatment failure. Outcome of UD-HSCT recipients is comparable to that of children receiving transplants from an HLA-identical sibling. PMID- 15353482 TI - Regulation of the single-chain urokinase-urokinase receptor complex activity by plasminogen and fibrin: novel mechanism of fibrin specificity. AB - Activation of plasminogen by urokinase plasminogen activator (uPA) plays important roles in several physiologic and pathologic conditions. Cells secrete uPA as a single-chain molecule (scuPA). scuPA can be activated by proteolytic cleavage to a 2-chain enzyme (tcuPA). scuPA is also activated when it binds to its receptor (uPAR). The mechanism by which the enzymatic activity of the scuPA/suPAR complex is regulated is only partially understood. We now report that the plasminogen activator activity of the scuPA/suPAR complex is inhibited by Glu and Lys-plasminogen, but not by mini-plasminogen. In contrast, neither Glunor Lys-plasminogen inhibits the activation of plasminogen by 2-chain uPA. Inhibition of scuPA/suPAR activity was evident at a Glu-plasminogen concentration of approximately 100 nM, and at physiologic plasma concentrations inhibition was nearly complete. A plasminogen fragment containing kringles 1-3 inhibited the enzymatic activity of scuPA/suPAR with an inhibition constant (Ki) equal to 1.9 microM, increased the Michaelis constant (Km) of scuPA/suPAR from 18 nM to 49 nM, and decreased the catalytic constant (Kcat) approximately 3-fold from 0.035 sec( 1) to 0.011 sec(-1). Inhibition of scuPA/suPAR by plasminogen was completely abolished in the presence of fibrin clots. These studies provide insight into the regulation of uPA-mediated plasminogen activation and identify a novel mechanism for its fibrin specificity. PMID- 15353483 TI - The Jab1/COP9 signalosome subcomplex is a downstream mediator of Bcr-Abl kinase activity and facilitates cell-cycle progression. AB - Jab1 is a multifunctional protein associated with the signaling pathway, cell cycle regulation, and development, and acts as a key subunit of COP9 signalosome (CSN). Jab1 promotes degradation of the cyclin-dependent kinase inhibitor p27(Kip1) by transportation from the nucleus to the cytoplasm. However, there has been no clear evidence for whether and how Jab1 contributes to malignant transformation in human cancers. Here we show that Bcr-Abl tyrosine kinase facilitates the down-regulation of p27 by modulating complex formation of Jab1/CSN through the mitogen-activated protein (MAP) kinase and phosphatidylinositol 3 (PI3) kinase signaling pathways. Nearly half of the chronic myelogenous leukemia cell lines and the murine hematopoietic precursor cells expressing Bcr-Abl exhibited a marked increase in the small loose Jab1 complex located in the cytoplasm. Inhibition of Bcr-Abl kinase by STI571 induced G1 arrest and caused a recovery of the p27 level with reduction of the small Jab1 complex from the cytoplasm. Either blockade of the MAP kinase and PI3 kinase pathways by specific inhibitors or Jab1 knockdown by small interfering RNA (siRNA) prevented p27 down-regulation as well as formation of the small complex. Thus, regulation of p27 via modulation of the Jab1 subcomplex is a novel mechanism whereby Bcr-Abl oncogenic signals accelerate abnormal cell proliferation. PMID- 15353484 TI - Splenic lymphoma with villous lymphocytes, associated with type II cryoglobulinemia and HCV infection: a new entity? AB - Hepatitis C virus (HCV) has been associated with the development of B-cell non Hodgkin lymphomas. We recently reported the regression of splenic lymphoma with villous lymphocytes (SLVL) in patients with HCV after antiviral treatment, demonstrating a direct role of HCV in lymphomagenesis. This study expands our previous results in 18 patients with chronic HCV and SLVL. Mixed cryoglobulinemia (MC) was present in all cases and was symptomatic in 13 (72%). All patients were treated with interferon alone or in association with ribavirin. Hematologic and virologic responses were correlated. Fourteen (78%) patients achieved a sustained complete hematologic response after clearance of HCV RNA. Two patients had a virologic partial response and achieved a complete hematologic response. Two virologic nonresponders achieved partial hematologic response. Regardless of the response, monoclonal immunoglobulin gene rearrangement persisted after treatment. This study underscores the role of HCV in the lymphomagenesis and the benefit of antiviral treatment for patients presenting with HCV-driven lymphoproliferations. PMID- 15353485 TI - High factor VIII levels in venous thromboembolism show linkage to imprinted loci on chromosomes 5 and 11. AB - High factor VIII (FVIII) levels are known to be a risk factor for deep venous thrombosis, but the mechanisms responsible for high FVIII levels remain unclear. Here, a new phenotype "FVIII level residuum" (FVIII-R) was defined in order to eliminate the impact of common determinants on FVIII levels. We studied 13 families of patients with thrombosis and reproducibly high FVIII levels of unknown origin. Since familial clustering was evident, we looked for a possible genetic basis. A genome scan was performed with 402 evenly spaced microsatellite markers. A quantitative linkage analysis using variance component methods showed suggestive evidence for linkage of FVIII-R with a locus on chromosome 8 (logarithm of odds [LOD] = 2.1). In addition, we performed parametric exploratory linkage analysis of dichotomized FVIII-R, taking a parent-of-origin effect into account. Single-trait-locus MOD-score analysis showed suggestive evidence for linkage under an imprinting model at chromosomes 5 and 11. Furthermore, a 2-trait locus analysis under a multiplicative model for the loci of chromosomes 5 and 11 yielded a remarkable LOD of 4.44. It confirmed the finding of paternal imprinting, obtained by single-trait-locus analysis, at both loci. Our results suggest that high FVIII levels in venous thromboembolism represent a complex trait caused by several genetic factors. PMID- 15353486 TI - Novel mechanism of G-CSF refractoriness in patients with severe congenital neutropenia. AB - Severe congenital neutropenia (SCN) is a rare disease diagnosed at or soon after birth, characterized by a myeloid maturation arrest in the bone marrow, ineffective neutrophil production, and recurrent infections. Most patients respond to treatment with granulocyte colony-stimulating factor (G-CSF), and the majority harbor mutations in the neutrophil elastase gene. In the subset of patients with SCN transforming to acute myeloid leukemia (AML), mutations that truncate the cytoplasmic tail of the G-CSF receptor (G-CSFR) have been detected. Here, we report a novel mutation in the extracellular portion of the G-CSFR within the WSXWS motif in a patient with SCN without AML who was refractory to G CSF treatment. The mutation affected a single allele and introduced a premature stop codon that deletes the distal extracellular region and the entire transmembrane and cytoplasmic portions of the G-CSFR. Expression of the mutant receptor in either myeloid or lymphoid cells was shown to alter subcellular trafficking of the wild-type (WT) G-CSFR by constitutively heterodimerizing with it. WT/mutant G-CSFR heterodimers appeared to be retained in the endoplasmic reticulum and/or Golgi and accumulate intracellularly. These findings together with 2 previous case reports of extracellular mutations in the G-CSFR in patients with SCN unresponsive to G-CSF suggest a common mechanism underlying G-CSF refractoriness. PMID- 15353487 TI - Combined adipocyte-macrophage fatty acid-binding protein deficiency improves metabolism, atherosclerosis, and survival in apolipoprotein E-deficient mice. AB - BACKGROUND: The adipocyte fatty acid-binding protein (FABP) aP2 is expressed by adipocytes and macrophages and modulates insulin resistance, glucose and lipid metabolism, and atherosclerosis. Insulin sensitivity is improved in obese but not in lean aP2-deficient mice. A second fatty acid-binding protein, mal1, also is expressed in adipocytes and macrophages, and mal1 deficiency produces similar effects on insulin resistance. We tested the hypothesis that combined aP2 and mal1 deficiency would produce synergistic effects on metabolism and reduce atherosclerosis in apolipoprotein E-deficient (apoE-/-) mice. METHODS AND RESULTS: Male and female apoE-/- mice null for both aP2 and mal1 (3KO) and apoE-/ controls were fed a low-fat chow diet for 16 or 56 weeks. Lean 3KO mice had significantly lower serum cholesterol and triglycerides as well as improved insulin and glucose tolerance as compared with controls. Analysis of atherosclerotic lesions in the 3KO mice showed dramatic reductions in both early (20 weeks) and late-stage (60 weeks) atherosclerosis. Strikingly, survival in the 3KO mice was improved by 67% as compared with apoE-/- controls when challenged with the Western diet for 1 year. CONCLUSIONS: Combined aP2 and mal1 deficiency improved glucose and lipid metabolism, reduced atherosclerosis, and improved survival in apoE-/- mice, making these proteins important therapeutic targets for the prevention of the cardiovascular consequences of the metabolic syndrome. PMID- 15353488 TI - Differential effects of apolipoprotein A-I-mimetic peptide on evolving and established atherosclerosis in apolipoprotein E-null mice. AB - BACKGROUND: Apolipoprotein (apo) A-I and apoA-I-mimetic peptides showed promise to prevent atherosclerosis development. Using a bypassed vein graft model in apoE null mice, we evaluated the effects of oral or intraperitoneal administration of an apoA-I-mimetic peptide on evolving atherosclerotic lesions in the vein graft and compared such effects on the established atherosclerotic lesions in aortic sinus in the same mice. METHODS AND RESULTS: We used apoE-null mice in which a segment of inferior vena cava was grafted into the right carotid artery at 16 weeks of age. Native aortic atherosclerotic lesions (established atherosclerosis) and vein-graft atherosclerotic lesions (evolving atherosclerosis) were assessed 4 weeks after daily oral (0.3 mg/mL) or intraperitoneal (50 microg in 200 microL saline) administration of an apoA-I-mimetic peptide, D4F. Mice receiving saline or water without D4F served as controls. Both oral and intraperitoneal administration of D4F reduced vein-graft atherosclerotic (evolving lesions) plaque size by 43% and 42%, plaque lipid by 70% and 49%, and macrophage immunoreactivity by 63% and 62%, respectively, compared with controls. In contrast, D4F had no effect on the native aortic sinus atherosclerotic lesions (established lesions). CONCLUSIONS: Oral and intraperitoneal administration of the apoA-I-mimetic peptide D4F significantly reduced rapidly evolving atherosclerotic lesions in vein grafts but not established atherosclerotic lesions in aortic sinus. These observations suggest that the type of atherosclerotic lesions and the time of initiation during the course of lesion evolution modulate the beneficial effects of apoA-I-mimetic peptides on atherosclerosis. PMID- 15353489 TI - Elevations in troponin I after percutaneous coronary interventions are associated with abnormal tissue-level perfusion in high-risk patients with non-ST-segment elevation acute coronary syndromes. AB - BACKGROUND: In the setting of non-ST-segment-elevation (NSTE) acute coronary syndromes (ACS), the pathophysiological mechanisms underlying post-percutaneous coronary intervention (PCI) cardiac troponin I (cTnI) elevation remain unclear. METHODS AND RESULTS: We evaluated the relationship between troponin elevation and tissue-level perfusion using the TIMI flow grade, corrected TIMI frame count, TIMI myocardial perfusion grade (TMPG), and myocardial contrast enhancement by intracoronary myocardial contrast echocardiography (MCE) before and immediately after PCI performed within 24 to 48 hours of hospital admission in 42 high-risk (angina at rest, unequivocal ST-segment depression, and cTnI elevation) patients with NSTE-ACS. All patients were treated with glycoprotein IIb/IIIa inhibitors (27 with tirofiban and 15 with abciximab) and had successful PCI. Fourteen patients had a postprocedural cTnI elevation, whereas 28 did not. TMPG 0/1 after PCI was observed more frequently in patients with postprocedural cTnI elevation (43% versus 7%; P<0.02). cTnI levels were higher among patients with TMPG 0/1 versus patients with TMPG 2/3 (5.3+/-2.7 versus 1.5+/-1.3 ng/mL; P<0.0001). Patients with postprocedural cTnI elevation also presented a significantly lower number of perfused segments at MCE (59% versus 81%; P=0.02) as well as a lower MCE score index (0.65+/-0.38 versus 0.89+/-0.21; P<0.02). CONCLUSIONS: Postprocedural cTnI elevation in high-risk patients with NSTE-ACS is associated with an abnormal tissue-level perfusion. PMID- 15353490 TI - Acute cardiovascular effects of fetal surgery in the human. AB - BACKGROUND: Prenatal surgery for congenital anomalies can prevent fetal demise or alter the course of organ development, resulting in a more favorable condition at birth. The indications for fetal surgery continue to expand, yet little is known about the acute sequelae of fetal surgery on the human cardiovascular system. METHODS AND RESULTS: Echocardiography was used to evaluate the heart before, during, and early after fetal surgery for congenital anomalies, including repair of myelomeningocele (MMC, n=51), resection of intrathoracic masses (ITM, n=15), tracheal occlusion for congenital diaphragmatic hernia (CDH, n=13), and resection of sacrococcygeal teratoma (SCT, n=4). Fetuses with MMC all had normal cardiovascular systems entering into fetal surgery, whereas those with ITM, CDH, and SCT all exhibited secondary cardiovascular sequelae of the anomaly present. At fetal surgery, heart rate increased acutely, and combined cardiac output diminished at the time of fetal incision for all groups including those with MMC, which suggests diminished stroke volume. Ventricular dysfunction and valvular dysfunction were identified in all groups, as was acute constriction of the ductus arteriosus. Fetuses with ITM and SCT had the most significant changes at surgery. CONCLUSIONS: Acute cardiovascular changes take place during fetal surgery that are likely a consequence of the physiology of the anomaly and the general effects of surgical stress, tocolytic agents, and anesthesia. Echocardiographic monitoring during fetal surgery is an important adjunct in the management of these patients. PMID- 15353491 TI - Omega-3 fatty acids in cardiac biopsies from heart transplantation patients: correlation with erythrocytes and response to supplementation. AB - BACKGROUND: Omega-3 fatty acids (FAs) appear to reduce the risk of sudden death from myocardial infarction. This reduction is believed to occur via the incorporation of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) into the myocardium itself, altering the dynamics of sodium and calcium channel function. The extent of incorporation has not been determined in humans. METHODS AND RESULTS: We first determined the correlation between red blood cell (RBC) and cardiac omega-3 FA levels in 20 heart transplant recipients. We then examined the effects of 6 months of omega-3 FA supplementation (1 g/d) on the FA composition of human cardiac and buccal tissue, RBCs, and plasma lipids in 25 other patients. Cardiac and RBC EPA+DHA levels were highly correlated (r=0.82, P<0.001). Supplementation increased EPA+DHA levels in cardiac tissue by 110%, in RBCs by 101%, in plasma by 139%, and in cheek cells by 73% (P<0.005 versus baseline for all; responses among tissues were not significantly different). CONCLUSIONS: Although any of the tissues examined could serve as a surrogate for cardiac omega 3 FA content, RBC EPA+DHA was highly correlated with cardiac EPA+DHA; the RBC omega-3 response to supplementation was similar to that of the heart; RBCs are easily collected and analyzed; and they have a less variable FA composition than plasma. Therefore, RBC EPA+DHA (also called the Omega-3 Index) may be the preferred surrogate for cardiac omega-3 FA status. PMID- 15353492 TI - Major vascular anomalies in Turner syndrome: prevalence and magnetic resonance angiographic features. AB - BACKGROUND: Turner syndrome (TS) is associated with aortic coarctation and dissection; hence, echocardiographic evaluation of all patients is currently recommended. X-ray angiography in clinically symptomatic patients has suggested a range of other vascular anomalies, but the true prevalence of such lesions in TS is unknown. To better understand the prevalence and pathogenesis of cardiovascular defects in TS, we prospectively evaluated a group of asymptomatic adult volunteers with TS using magnetic resonance (MR) angiography. METHODS AND RESULTS: A total of 85 adults with TS and 27 normal female adult volunteers underwent gadolinium-enhanced 3D MR angiography. A high prevalence of aortic anomalies was seen in women with TS, including elongation of the transverse arch (49%), aortic coarctation (12%), and aberrant right subclavian artery (8%). Venous anomalies were also prominent, including persistent left superior vena cava (13%) and partial anomalous pulmonary venous return (13%). None of these anomalies were found in healthy female controls. The constellation of elongation of the transverse arch, aortic coarctation, and persistent left superior vena cava was significantly associated with women with TS. Neck webbing and increased thoracic anterior-to-posterior dimension diameters were strong predictors for arterial and venous anomalies. CONCLUSIONS: Thoracic vascular anomalies are common in TS, occurring in approximately 50% of a group not preselected for cardiovascular disease. The highly significant association between neck webbing, increased chest diameter, and these vascular anomalies suggests that in utero, centrally localized lymphatic obstruction may contribute to these cardiovascular deformities in TS. Improved recognition of these often-undetected vascular lesions may be important for identification of patients in need of closer cardiovascular monitoring. PMID- 15353493 TI - Upper-extremity deep vein thrombosis: a prospective registry of 592 patients. AB - BACKGROUND: Upper-extremity deep vein thrombosis (UEDVT) occurs spontaneously or sometimes develops as a complication of pacemaker use, long-term use of a central venous catheter (CVC), or cancer. METHODS AND RESULTS: To improve our understanding of UEDVT, we compared the demographics, symptoms, risk factors, prophylaxis, and initial management of 324 (6%) patients with central venous catheter (CVC)-associated UEDVT, 268 (5%) patients with non-CVC-associated UEDVT, and 4796 (89%) patients with lower-extremity DVT from a prospective US multicenter DVT registry. The non-CVC-associated UEDVT patients were younger (59.2+/-18.2 versus 64.2+/-16.9 years old; P<0.0001), less often white (65% versus 73%; P<0.01), leaner (body mass index [BMI] 26.8+/-7.1 versus 28.5+/-7.3 kg/m2; P<0.001), and more likely to smoke (19% versus 13%; P=0.02) than the lower extremity DVT patients. By way of propensity analysis and multivariable logistic regression analysis, we determined that an indwelling CVC was the strongest independent predictor of UEDVT (odds ratio [OR], 7.3; 95% confidence interval [CI], 5.8 to 9.2). An age of <67 years, a BMI of <25 kg/m2, and hospitalization were the independent predictors of non-CVC-associated UEDVT. Most (68%) UEDVT patients were evaluated while they were inpatients. Only 20% of the 378 UEDVT patients who did not have an obvious contraindication to anticoagulation received prophylaxis at the time of diagnosis. CONCLUSIONS: UEDVT risk factors differ from the conventional risk factors for lower-extremity DVT. Our findings identify deficiencies in our current understanding and the prophylaxis of UEDVT and generate hypotheses for future research efforts. PMID- 15353494 TI - Interleukin-1 receptor signaling mediates atherosclerosis associated with bacterial exposure and/or a high-fat diet in a murine apolipoprotein E heterozygote model: pharmacotherapeutic implications. AB - BACKGROUND: Current data demonstrate that progressive atherosclerosis is associated with activation of the inflammatory process, as evidenced by systemic elevations of molecules such as tumor necrosis factor, interleukin (IL)-6, and IL 1. It has been postulated that inflammatory events within an atherogenic lesion are induced by oxidized LDL. Recent evidence suggests that infectious agents, including those that cause periodontal disease, may also play an important role. Studies presented here tested the hypothesis that IL-1 receptor (IL-1R1) signaling plays a crucial role in bacteria- and/or high-fat diet (HFD)-enhanced atherogenesis. METHODS AND RESULTS: Ten-week-old ApoE+/- mice lacking either 1 IL 1R1 allele (ApoE+/-/IL-1R1+/-) or 2 IL-1R1 alleles (ApoE+/-/IL-1R1-/-) fed either an HFD or regular chow were inoculated intravenously with live Porphyromonas gingivalis (P gingivalis) (10(7) CFU), an important periodontal pathogen, or vehicle once per week for 14 or 24 consecutive weeks. Histomorphometry of plaque cross-sectional area in the proximal aortas, en face measurement of plaque area over the aortic trees, and ELISA for systemic proinflammatory mediators were performed. Atherosclerotic lesions of proximal aortas and aortic tree were substantially reduced in ApoE+/-/IL-1R1-/- mice than in ApoE+/-/IL-1R1+/- mice challenged with P gingivalis. At 24 weeks after P gingivalis inoculation, proximal aortic lesion size quantified by histomorphometry was 5-fold-reduced in chow-fed ApoE+/-/IL-1R1-/- mice than in ApoE+/-/IL-1R1+/- mice (P<0.05). In the HFD group, ApoE+/-/IL-1R1-/- mice exhibited marked attenuation of the progression of atherosclerotic lesions (78% to 97%), with and without P gingivalis inoculation (P<0.05). CONCLUSIONS: Ablation of IL-1R1 under P gingivalis challenge and/or an HFD reduced the progression of atherosclerotic plaques. These results indicate that IL-1 plays a crucial role in bacteria- and/or HFD-enhanced atherogenesis. PMID- 15353495 TI - Estrogen modulates inflammatory mediator expression and neutrophil chemotaxis in injured arteries. AB - BACKGROUND: We have previously shown that estrogen (17beta-estradiol; E2) inhibits neointima formation and migration of leukocytes, particularly neutrophils, into rat carotid arteries after acute endoluminal injury. This study tested the hypothesis that E2 inhibits expression of adhesion molecules, chemokines, and proinflammatory cytokines in rat carotid arteries in the early hours after balloon injury, thus attenuating the stimulus for leukocyte entry and negatively modulating the injury response. METHODS AND RESULTS: Ovariectomized (OVX) rats were randomly assigned to treatment with E2 or vehicle (V) and subjected to balloon injury of the right carotid artery. After 2, 6, and 24 hours, rats were euthanized, and both carotid arteries were processed for real time reverse transcription-polymerase chain reaction (2 and 24 hours), ELISA (6 hours), or neutrophil chemotaxis assay (24 hours). Expression of mRNA for adhesion molecules (P-selectin, vascular cell adhesion molecule-1, and intercellular adhesion molecule-1), chemoattractants (cytokine-induced neutrophil chemoattractant [CINC]-2beta and monocyte chemoattractant protein [MCP]-1), and proinflammatory cytokines (interleukin [IL]-1 and IL-6) was markedly increased (2 to 5000 times) in injured arteries of OVX+V rats at 2 hours and was reduced by 24 hours. E2 significantly attenuated expression of the proinflammatory mediators (by 60% to 80%) at 2 hours. ELISA confirmed injury-induced upregulation of neutrophil and monocyte/macrophage chemoattractants (CINC-2alpha, MCP-1) in OVX+V arteries and E2-induced inhibition of CINC-2alpha expression. E2 significantly (by 65%) inhibited neutrophil chemotactic activity of arterial homogenates. CONCLUSIONS: E2 attenuates the early vascular injury response, at least in part, by negatively modulating proinflammatory mediator expression and the resultant chemotactic activity of injured vessels for neutrophils. PMID- 15353496 TI - Value of delayed-enhancement cardiovascular magnetic resonance imaging in predicting myocardial viability after surgical revascularization. AB - BACKGROUND: Despite the accepted utility of delayed-enhancement MRI in identifying irreversible myocardial injury, no study has yet assessed its role as a viability tool exclusively in the setting of coronary artery bypass surgery (CABG), and no study has repeated delayed-enhancement MRI late after revascularization. In a clinical trial in which patients underwent CABG by either the off-pump or on-pump surgical technique, we hypothesized that (1) preoperative delayed-enhancement MRI would have high diagnostic accuracy in predicting viability and (2) the occurrence of perioperative myocardial necrosis would affect late regional wall motion recovery. METHODS AND RESULTS: Fifty-two patients undergoing multivessel CABG were studied by preoperative and early (day 6) and late (6 months) postoperative cine MRI for global and regional functional assessment and delayed-enhancement MRI for assessment of irreversible myocardial injury. Preoperatively, 611 segments (21%) had abnormal regional function, whereas 421 segments (14%) showed evidence of hyperenhancement. At 6 months after revascularization, 57% (343 of 611) of dysfunctional segments improved contraction by at least 1 grade. When all preoperative dysfunctional segments were analyzed, there was a strong correlation between the transmural extent of hyperenhancement and the recovery in regional function at 6 months (P<0.001). Of a total of 96 previously dysfunctional but nonenhancing or minimally hyperenhancing myocardial segments that did not improve regional function at 6 months, 35 (36%) demonstrated new perioperative hyperenhancement in the early postoperative MRI scan. CONCLUSIONS: Delayed-enhancement MRI is a powerful predictor of myocardial viability after surgery, suggesting an important role for this technique in clinical viability assessment. PMID- 15353497 TI - Effect of ramipril in reducing sudden deaths and nonfatal cardiac arrests in high risk individuals without heart failure or left ventricular dysfunction. AB - BACKGROUND: ACE inhibitor therapy reduces the risk of cardiovascular death, myocardial infarction, stroke, hospitalization for heart failure, and need for revascularization in high-risk patients with clinical heart failure, overt left ventricular systolic dysfunction, or vascular disease. In patients with clinical heart failure or overt left ventricular systolic dysfunction, ACE inhibitor therapy also reduces the risk of sudden or arrhythmia-related cardiac death. The objective of this study was to assess the effect of the ACE inhibitor ramipril on sudden unexpected death or resuscitated cardiac arrest among the 9297 individuals without clinical heart failure or overt left ventricular dysfunction enrolled in the Heart Outcomes Prevention Evaluation (HOPE) trial. METHODS AND RESULTS: During the median follow-up of 4.5 years, the composite outcome of unexpected death, documented arrhythmic death, or resuscitated cardiac arrest was reduced by 21% in patients randomized to ramipril therapy compared with those randomized to placebo. There were 155 (3.3%) composite outcome events in patients randomized to ramipril therapy compared with 195 (4.2%) such events in patients randomized to placebo (RR 0.79, 95% CI 0.64 to 0.98, P=0.028). There were trends toward reductions in fatal primary outcome events (unexpected death or documented arrhythmic death; RR 0.81, 95% CI 0.64 to 1.02, P=0.072) and in nonfatal primary outcome events (resuscitated cardiac arrest; RR 0.65, 95% CI 0.38 to 1.13, P=0.127) in the ramipril treatment group. CONCLUSIONS: Ramipril reduces the risk of fatal and nonfatal serious arrhythmic events in high-risk patients without clinical heart failure or overt left ventricular systolic dysfunction. PMID- 15353498 TI - High-dose atorvastatin reduces total plasma levels of oxidized phospholipids and immune complexes present on apolipoprotein B-100 in patients with acute coronary syndromes in the MIRACL trial. AB - BACKGROUND: Oxidized phospholipids (OxPL) are present within atherosclerotic plaques and bound by lipoprotein (a) [Lp(a)] in plasma. This study evaluated the impact of atorvastatin on oxidized LDL (OxLDL) in patients with acute coronary syndromes (ACS). METHODS AND RESULTS: OxLDL-E06 (OxPL content on apolipoprotein B 100 [apoB] detected by antibody E06), apoB-100 immune complexes (apoB-IC), OxLDL autoantibodies, and Lp(a) levels were measured in 2341 patients at baseline and after 16 weeks of treatment with atorvastatin 80 mg/d or placebo. The OxLDL-E06 and apoB-IC data are reported per apoB-100 particle (OxPL/apoB, IC/apoB) and as total levels on all apoB-100 particles (total apoB-OxPL and total apoB-IC [eg, OxPL/apoB or IC/apoBxapoB-100 levels]). Compared with baseline values, atorvastatin reduced apoB-100 (-33%), total apoB-OxPL (-29.7%), total apoB-IC IgG (-29.5%), and IgM (-25.7%) (P<0.0001 for all), whereas no change or an increase was observed with placebo. When normalized per apoB-100, compared with placebo, atorvastatin increased OxPL/apoB (9.5% versus -3.9%, P<0.0001) and Lp(a) (8.8% versus -0.7%, (P<0.0001). A strong correlation was noted between OxPL/apoB and Lp(a) (R=0.85, P<0.0001), consistent with previous data that Lp(a) binds OxPL. CONCLUSIONS: After atorvastatin treatment, total OxPL on all apoB-100 particles was decreased. However, there was enrichment of OxPL on a smaller pool of apoB 100 particles, in parallel with similar increases in Lp(a), suggesting binding by Lp(a). These data support the hypothesis that atorvastatin promotes mobilization and clearance of proinflammatory OxPL, which may contribute to a reduction in ischemic events after ACS. PMID- 15353499 TI - Predictors of heart failure among women with coronary disease. AB - BACKGROUND: Although heart failure is common among women with coronary disease, the risk factors for developing heart failure have not been well studied. We determined the risk factors for developing heart failure among postmenopausal women with established coronary disease. METHODS AND RESULTS: This is a prospective cohort study using data from the Heart and Estrogen/progestin Replacement Study (HERS), a randomized, blinded, placebo-controlled trial of 4.1 years' duration, and subsequent open-label observational follow-up for 2.7 years (HERS II), performed at 20 US clinical centers between 1993 and 2000. Of the 2763 postmenopausal women with established coronary disease in the HERS trial, we studied the 2391 women with no heart failure at baseline by self-report and physical examination. The primary outcome of this analysis was incident heart failure defined by hospital admission or death from heart failure. During the 6.3+/-1.4-year follow-up, 237 women (10%) developed heart failure. Nine predictors were identified: diabetes (defined as a self-reported history of diabetes on treatment), atrial fibrillation, myocardial infarction, creatinine clearance <40 mL/min, systolic blood pressure >120 mm Hg, current smoking, body mass index >35 kg/m2, left bundle-branch block, and left ventricular hypertrophy. Randomization to estrogen/progestin was not associated with heart failure (hazard ratio=1.0; 95% CI, 0.7 to 1.3). Diabetes was the strongest risk factor (adjusted hazard ratio=3.1; 95% CI, 2.3 to 4.2). Diabetic women with elevated body mass index or depressed creatinine clearance were at highest risk, with annual incidence rates of 7% and 13%, respectively. Among diabetic women, hyperglycemia was associated with heart failure risk (adjusted hazard ratio=3.0; 95% CI, 1.2 to 7.5 for fasting glucose >300 mg/dL compared with fasting glucose 80 to 150 mg/dL). CONCLUSIONS: We identified 9 predictors of heart failure in postmenopausal women with coronary disease. Diabetes was the strongest risk factor, particularly when poorly controlled or with concomitant renal insufficiency or obesity. PMID- 15353500 TI - Disruption of endothelial-cell caveolin-1alpha/raft scaffolding during development of monocrotaline-induced pulmonary hypertension. AB - BACKGROUND: In the monocrotaline (MCT)-treated rat, there is marked stimulation of DNA synthesis and megalocytosis of pulmonary arterial endothelial cells (PAECs) within 3 to 4 days, followed by pulmonary hypertension (PH) 10 to 14 days later. Growing evidence implicates caveolin-1 (cav-1) in plasma membrane rafts as a negative regulator of promitogenic signaling. We have investigated the integrity and function of endothelial cell-selective cav-1alpha/raft signaling in MCT-induced PH. METHODS AND RESULTS: Although PH and right ventricular hypertrophy developed by 2 weeks after MCT, a reduction in cav-1alpha levels in the lung was apparent within 48 hours, declining to approximately 30% by 2 weeks, accompanied by an increase in activation of the promitogenic transcription factor STAT3 (PY-STAT3). Immunofluorescence studies showed a selective loss of cav 1alpha and platelet endothelial cell adhesion molecule-1 in the PAEC layer within 48 hours after MCT but an increase in PY-STAT3. PAECs with cav-1alpha loss displayed high PY-STAT3 and nuclear immunostaining for proliferating cell nuclear antigen (PCNA). Biochemical studies showed a loss of cav-1alpha from the detergent-resistant lipid raft fraction concomitant with hyperactivation of STAT3. Moreover, cultured PAECs treated with MCT-pyrrole for 48 hours developed megalocytosis associated with hypo-oligomerization and reduction of cav-1alpha, hyperactivation of STAT3 and ERK1/2 signaling, and stimulation of DNA synthesis. CONCLUSIONS: MCT-induced disruption of cav-1alpha chaperone and scaffolding function in PAECs likely accounts for diverse alterations in endothelial cell signaling in this model of PH. PMID- 15353501 TI - Mechanisms of organized left atrial tachycardias occurring after pulmonary vein isolation. AB - BACKGROUND: A proarrhythmic consequence of pulmonary vein (PV) isolation can be a recurrent organized left atrial (LA) tachycardia after ablation. This arrhythmia is frequently referred to as "left atrial flutter," but the mechanism and best ablation strategy have not been determined. METHODS AND RESULTS: Isolation of arrhythmogenic PVs was initially performed by segmental ostial PV ablation guided by a circular mapping catheter in 341 patients. Patients whose predominant recurrent arrhythmia was a persistent organized tachycardia returned for mapping and ablation. Recurrent organized LA tachycardias (cycle length 253+/-33 ms, range 213 to 328 ms) occurred in 10 (2.9%) of 341 patients (age 59+/-9 years, 1 woman). Mapping was consistent with a focal origin in 8 patients and with macroreentry in 1 patient and was unclear in 1 patient owing to degeneration to atrial fibrillation. Focal tachycardias originated from reconnected segments of prior isolated PVs (6 patients), the posterior LA (1 patient), or the superior septum (1 patient). Focal atrial tachycardias were ablated with point lesions that targeted the earliest activation. All reconnected PVs were also reisolated. Reentrant LA flutter occurred around the left PVs in 1 patient. After 6.7+/-2.3 months of follow-up, 9 (90%) of 10 patients were arrhythmia free (4 of whom were taking antiarrhythmic drug therapy), and one was having recurrent atrial fibrillation. CONCLUSIONS: Recurrent organized LA tachycardia after PV isolation is uncommon and typically has a focal origin from reconnected PV ostia. Reisolation of the PV and ablation of non-PV foci are sufficient to treat this proarrhythmia. Linear lesions are only required when a macroreentrant mechanism is present. PMID- 15353502 TI - Plasma level of B-type natriuretic peptide as a prognostic marker after acute myocardial infarction: a long-term follow-up analysis. AB - BACKGROUND: Circulating levels of B-type natriuretic peptide (BNP), a cardiac hormone, reflect the severity of cardiac dysfunction. Because the plasma BNP level changes dramatically during the period after the onset of acute myocardial infarction (AMI), identification of a suitable sampling time is problematic. There have been several reports indicating that the plasma BNP level obtained in the acute phase of AMI can be used as a prognostic marker. We examined whether the plasma BNP level measured 3 to 4 weeks after the onset of AMI represents a reliable prognostic marker for patients with AMI. METHODS AND RESULTS: We analyzed 145 consecutive patients with AMI. Plasma BNP levels were measured during the 3 to 4 weeks after onset of AMI. Of those patients, 23 experienced fatal cardiac events during this study. The mean follow-up period was 58.6 months. Log BNP, left ventricular end-diastolic pressure, and pulmonary vascular resistance were all significantly higher in the cardiac death group, and there were more men and more patients with a history of heart failure in the cardiac death group. A Cox proportional hazards model analysis showed that log BNP was an independent predictor of cardiac death. The survival rate was significantly higher in patients with log BNP <2.26 (180 pg/mL) than in those with log BNP > or =2.26. CONCLUSIONS: The plasma BNP level obtained 3 to 4 weeks after the onset of AMI can be used as an independent predictor of cardiac death in patients with AMI. PMID- 15353503 TI - Systemic inhibition of nitric oxide synthase unmasks neural constraint of maximal myocardial blood flow in humans. AB - BACKGROUND: Nitric oxide (NO) is an endothelial mediator that regulates vascular smooth muscle tone, but it may exert its cardiovascular action also by modulating the autonomic control of vasomotor tone. We assessed the effect of simultaneous inhibition of both endothelial (eNOS) and neuronal (nNOS) NO synthase isoforms on myocardial blood flow (MBF) and coronary flow reserve (CFR) in volunteers and in (denervated) transplant recipients. METHODS AND RESULTS: MBF (mL x min(-1) x g( 1)) was measured at rest and during adenosine-induced hyperemia with positron emission tomography and 15O-labeled water. CFR was calculated as adenosine/resting MBF. Measurements were repeated during one of the following intravenous infusions: group 1 (n=12), saline; group 2 (n=9), 3 mg/kg N(G) monomethyl-L-arginine (L-NMMA), which crosses the blood-brain barrier and inhibits both eNOS and nNOS; group 3 (n=13), 10 mg/kg L-NMMA; group 4 (n=8), phenylephrine titrated to simulate the hemodynamic changes in group 3; and group 5 (n=6), 10 mg/kg L-NMMA infused into the heart transplant recipients. After intervention, hyperemic MBF and CFR were unchanged in groups 1, 2, and 4. By contrast, both hyperemic MBF (+53%, P<0.0001 versus baseline) and CFR (+52%, P<0.0001 versus baseline) increased in group 3, whereas they remained unchanged in group 5, which suggests that an intact cardiac innervation was required for the increase in MBF and CFR observed in group 3. CONCLUSIONS: The results of the present study suggest that maximal adenosine-induced hyperemia and CFR in humans are constrained by neurally mediated vasoconstriction, which can be relieved by systemic NOS inhibition with L-NMMA. PMID- 15353504 TI - Oxygen-sensitive Kv channel gene transfer confers oxygen responsiveness to preterm rabbit and remodeled human ductus arteriosus: implications for infants with patent ductus arteriosus. AB - BACKGROUND: Oxygen (O2)-sensitive K+ channels mediate acute O2 sensing in many tissues. At birth, initial functional closure of the ductus arteriosus (DA) results from O2-induced vasoconstriction. This mechanism often fails in premature infants, resulting in persistent DA, a common form of congenital heart disease. We hypothesized that the basis for impaired O2 constriction in preterm DA is reduced expression and function of O2-sensitive, voltage-gated (Kv) channels. METHODS AND RESULTS: Preterm rabbit DA rings have reduced O2 constriction (even after inhibition of prostaglandin and nitric oxide synthases), and preterm DA smooth muscle cells (DASMCs) display reduced O2-sensitive K+ current. This is associated with decreased mRNA and protein expression of certain O2-sensitive Kv channels (Kv1.5 and Kv2.1) but equivalent expression of the L-type calcium channel. Transmural Kv1.5 or Kv2.1 gene transfer "rescues" the developmental deficiency, conferring O2 responsiveness to preterm rabbit DAs. Targeted SMC Kv1.5 gene transfer also enhances O2 constriction in human DAs. CONCLUSIONS: These data demonstrate a central role for developmentally regulated DASMC O2 sensitive Kv channels in the functional closure of the DA. Modulation of Kv channels may have therapeutic potential in diseases associated with impaired O2 responsiveness, including persistent DA. PMID- 15353505 TI - Determination of human ventricular repolarization by noncontact mapping: validation with monophasic action potential recordings. AB - BACKGROUND: Noncontact mapping (NCM) has not been validated as a clinical technique to measure ventricular repolarization. We used NCM to determine repolarization characteristics by analysis of reconstructed unipolar electrograms (UEs) at the same sites as monophasic action potential (MAP) recordings in the human ventricle. METHODS AND RESULTS: MAPs were recorded from a total of 355 beats at 46 sites in the left or right ventricle of 9 patients undergoing ablation of ventricular tachycardia guided by NCM (EnSite system). Measurements were made during sinus rhythm, constant right ventricular pacing, and ventricular extrastimuli during restitution-curve construction. The EnGuide locator signal was used to document MAP catheter locations on the endocardial geometry. UE determined activation-recovery interval (ARI) measured at the maximum derivative of the T wave (Wyatt method) and the minimum derivative of the positive T wave (alternative method) was correlated with MAP measured at 90% repolarization (MAP90%) at the same sites. ARI correlated with MAP90% during steady state by the Wyatt method (r=0.83, P<0.001) and the alternative method (r=0.94, P<0.001). Restitution curves constructed from MAP and UE data exhibited the same characteristics, with a mean correlation coefficient of 0.95 (range, 0.90 to 0.99, P<0.001). The error between ARI and MAP90% was greater over a shorter diastolic coupling interval but was not influenced by distance of the sampling site from the multielectrode array. CONCLUSIONS: NCM accurately determines steady state and dynamic endocardial repolarization in humans. Global, high-density, NCM data could be used to characterize abnormalities of human ventricular repolarization. PMID- 15353506 TI - Outcome in elderly patients undergoing primary coronary intervention for acute myocardial infarction: results from the Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications (CADILLAC) trial. AB - BACKGROUND: Biological age is a strong determinant of prognosis in patients with acute myocardial infarction (AMI). We sought to examine the impact of age after primary percutaneous coronary intervention in AMI and to determine whether routine coronary stent implantation and/or platelet glycoprotein IIb/IIIa inhibitors improve clinical outcomes in elderly patients after primary angioplasty. METHODS AND RESULTS: In the CADILLAC trial, 2082 patients with AMI were randomized to balloon angioplasty, angioplasty plus abciximab, stenting alone, or stenting plus abciximab. No patient was excluded on the basis of advanced age; patients ranging from 21 to 95 years of age were enrolled. One-year mortality increased for each decile of age, exponentially after 65 years of age (1.6% for patients <55 years, 2.1% for 55 to 65 years, 7.1% for 65 to 75 years, 11.1% for patients >75 years; P<0.0001). Elderly patients also had increased rates of stroke and major bleeding compared with their younger counterparts. Among elderly patients (> or =65 years), 1-year rates of ischemic target revascularization (7.0% versus 17.6%; P<0.0001) and subacute or late thrombosis (0% versus 2.2%; P=0.005) were reduced with stenting compared with balloon angioplasty. Routine abciximab administration, although safe, was not of definite benefit in elderly patients. Rates of mortality, reinfarction, disabling stroke, and major bleeding in the elderly were independent of reperfusion modality. CONCLUSIONS: Despite contemporary mechanical reperfusion strategies, mortality, major bleeding, and stroke rates remain high in elderly patients undergoing primary percutaneous coronary intervention, outcomes that are not affected by stents or glycoprotein IIb/IIIa inhibitors. By reducing restenosis, however, stent implantation improves clinical outcomes in elderly patients with AMI. PMID- 15353507 TI - Effectiveness of beta-blockade in experimental chronic aortic regurgitation. AB - BACKGROUND: Past studies have suggested that the adrenergic system becomes abnormally activated in chronic volume overload, such as in severe aortic valve regurgitation (AR). However, the effectiveness of agents directed against this adrenergic activation has never been adequately tested in chronic AR. We therefore tested the effects of metoprolol treatment on the left ventricular (LV) function and remodeling in severe chronic AR in rats. METHODS AND RESULTS: Severe AR was created in adult male Wistar rats by retrograde puncture of the aortic leaflets under echocardiographic guidance. Two weeks later, some animals received metoprolol treatment (25 mg/kg) orally for 24 weeks, and some were left untreated. LV dimensions, ejection fraction, and filling parameters were evaluated by echocardiography. Hearts were harvested at 1, 2, 14, and 180 days for the evaluation of hypertrophy, beta-adrenergic receptor status, and extracellular matrix remodeling. We found that metoprolol treatment prevented LV dilatation and preserved the ejection fraction and filling parameters compared with untreated animals. Metoprolol increased the expression of beta1 adrenoreceptor mRNA and reduced G protein receptor kinase 2 levels. Collagen I and III mRNA levels were reduced. Cardiac myocyte hypertrophy was also prevented. CONCLUSIONS: In our experimental model of severe AR, metoprolol treatment had a significant beneficial global effect on LV remodeling and function. These results suggest that the adrenergic system is important in the development of volume overload cardiomyopathy in AR and that adrenergic-blocking agents may play a role in the treatment of this disease. PMID- 15353508 TI - Transplacental fetal treatment improves the outcome of prenatally diagnosed complete atrioventricular block without structural heart disease. AB - BACKGROUND: Untreated isolated fetal complete atrioventricular block (CAVB) has a significant mortality rate. A standardized treatment approach, including maternal dexamethasone at CAVB diagnosis and beta-stimulation for fetal heart rates <55 bpm, has been used at our institutions since 1997. The study presents the impact of this approach. METHODS AND RESULTS: Thirty-seven consecutive cases of fetal CAVB since 1990 were studied. Mean age at diagnosis was 25.6+/-5.2 gestational weeks. In 33 patients (92%), CAVB was associated with maternal anti-Ro/La autoantibodies. Patients were separated into those diagnosed between 1990 and 1996 (group 1; n=16) and those diagnosed between 1997 and 2003 (group 2; n=21). The 2 study groups were comparable in the clinical presentation at CAVB diagnosis but did differ in prenatal management (treated patients: group 1, 4/16; group 2, 18/21; P<0.0001). Overall, 22 fetuses were treated, 21 with dexamethasone and 9 with beta-stimulation for a mean of 7.5+/-4.5 weeks. Live-birth and 1-year survival rates of group 1 were 80% and 47%, and these improved to 95% for group 2 patients (P<0.01). The 21 patients treated with dexamethasone had a 1-year survival rate of 90%, compared with 46% without glucocorticoid therapy (P<0.02). Immune-mediated conditions (myocarditis, hepatitis, cardiomyopathy) resulting in postnatal death or heart transplantation were significantly more common in untreated anti-Ro/La antibody-associated pregnancies compared with patients treated with steroids (0/18 versus 4/9 live births; P=0.007). CONCLUSIONS: A standardized treatment approach, including transplacental fetal administration of dexamethasone and beta-stimulation at heart rates <55 bpm, reduced the morbidity and improved the outcome of isolated fetal CAVB. PMID- 15353509 TI - In vivo 16-slice, multidetector-row computed tomography for the assessment of experimental atherosclerosis: comparison with magnetic resonance imaging and histopathology. AB - BACKGROUND: Noninvasive imaging can detect early atherosclerotic disease. Magnetic resonance imaging (MRI), because of its excellent spatial resolution, is already established as a tool for plaque characterization. Sixteen-slice, multidetector-row computed tomography (MDCT) was recently introduced into the field of cardiac imaging, with promising results for noninvasive angiography. We compared the capabilities of MDCT and MRI for the assessment of noncalcified, atherosclerotic plaques. METHODS AND RESULTS: Six atherosclerotic rabbits underwent in vivo imaging by MDCT and 1.5-T MRI. MDCT parameters were 120 kV, 120 mA/s, collimation 12x0.75, and spatial resolution 0.6x0.6 mm. MRI parameters were as follows: for proton density, repetition time/echo time (TR/TE) 2300/5.6; for T2, TR/TE 2300/62; and for T1, TR/TE 800/5.6; slice thickness was 3 mm and spatial resolution, 0.3x0.3 mm. Blinded analysis of 3-mm axial reconstructions from MDCT and the carefully matched MRI images (182 sections) showed excellent agreement between both modalities. MDCT yielded a slightly larger lumen area, anteroposterior diameters, and lateral diameters, with no significant differences in total vessel area. The sensitivity and specificity, respectively, to detect noncalcified, atherosclerotic plaques were 89% and 77% for MDCT and 97% and 94% for MRI. Fibrous-rich and lipid-rich plaque could not be differentiated visually, although they showed different attenuation properties (116+/-27 vs 51+/-25 Hounsfield units, P<0.01). CONCLUSIONS: Both techniques allow reliable detection of noncalcified, atherosclerotic plaques and accurate assessment of vessel areas and diameters. MDCT offers the additive value of a very short image acquisition time when compared with MRI. The subtle measurement differences found between modalities may be due to the better spatial resolution of MRI, which probably explains its superiority for tissue characterization. PMID- 15353510 TI - Exogenous thioredoxin reduces inflammation in autoimmune myocarditis. PMID- 15353511 TI - Targeted therapy to prevent progression of calcific aortic stenosis. PMID- 15353512 TI - Images in cardiovascular medicine. Multiple radial artery plaques. PMID- 15353513 TI - Images in cardiovascular medicine. Multimodality imaging of hemangiectasia hypertrophicans (Klippel-Trenaunay-Parkes-Weber syndrome). PMID- 15353514 TI - Inhibition of Rho-kinase in the brainstem augments baroreflex control of heart rate in rats. AB - The Rho/Rho-kinase pathway in the nucleus tractus solitarii (NTS) of the brain stem contributes to blood pressure regulation. Activation of this pathway might be involved in the central nervous system mechanisms of hypertension. The aim of the present study was to determine whether baroreflex control of heart rate is altered by inhibition of Rho-kinase in the NTS. Adenovirus vectors encoding dominant-negative Rho-kinase or beta-galactosidase were transfected into the nucleus tractus solitarii of Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). Baroreflex control of heart rate was examined by changing arterial pressure with an intravenous infusion of phenylephrine or sodium nitroprusside. The maximum gain of baroreflex control of heart rate was attenuated in SHR compared with WKY before the gene transfer. Transfection of adenovirus vectors encoding dominant-negative Rho-kinase significantly augmented the maximum gain in both WKY and SHR. The extent of this augmentation, however, was greater in SHR than in WKY. After treatment with metoprolol, the maximum gain was significantly decreased in rats transfected with adenovirus vectors encoding dominant-negative Rho-kinase, but not in nontransfected rats. In contrast, after treatment with atropine, the maximum gain was greater in rats transfected with adenovirus vectors encoding dominant-negative Rho-kinase compared with nontransfected rats, although it was decreased in both groups. These results suggest that inhibition of Rho-kinase in the NTS augments baroreflex control of heart rate, in both WKY and SHR, probably because of a cardiac sympathoinhibitory effect. PMID- 15353515 TI - Summary of the Fourth Report on the Diagnosis, Evaluation, and Treatment of High Blood Pressure in Children and Adolescents. PMID- 15353516 TI - A piece of my mind. In memoriam. PMID- 15353517 TI - Asia's growing HIV/AIDS epidemics in spotlight at International Conference. PMID- 15353518 TI - New HIV prevention strategies urged: averting new infections key to controlling pandemic. PMID- 15353519 TI - Physicians advised on how to offer pain relief while preventing opioid abuse. PMID- 15353520 TI - Improvement seen in US immunization rates. PMID- 15353524 TI - The AMA health insurance proposal. PMID- 15353525 TI - The AMA health insurance proposal. PMID- 15353526 TI - Corpulence and carbs in 1892. PMID- 15353527 TI - Parental atrial fibrillation as a risk factor for atrial fibrillation in offspring. PMID- 15353528 TI - Staging for antiretroviral therapy among HIV-infected drug users. PMID- 15353529 TI - Incidence of HIV superinfection following primary infection. PMID- 15353530 TI - Relationship of physical fitness vs body mass index with coronary artery disease and cardiovascular events in women. AB - CONTEXT: Individual contributions of obesity and physical fitness (physical activity and functional capacity) to risk of coronary heart disease in women remain unclear. OBJECTIVE: To investigate the relationships of measures of obesity (body mass index [BMI], waist circumference, waist-hip ratio, and waist height ratio) and physical fitness (self-reported Duke Activity Status Index [DASI] and Postmenopausal Estrogen-Progestin Intervention questionnaire [PEPI-Q] scores) with coronary artery disease (CAD) risk factors, angiographic CAD, and adverse cardiovascular (CV) events in women evaluated for suspected myocardial ischemia. DESIGN, SETTING, AND PARTICIPANTS: The National Heart, Lung, and Blood Institute-sponsored Women's Ischemia Syndrome Evaluation (WISE) is a multicenter prospective cohort study. From 1996-2000, 936 women were enrolled at 4 US academic medical centers at the time of clinically indicated coronary angiography and then assessed (mean follow-up, 3.9 [SD, 1.8] years) for adverse outcomes. MAIN OUTCOME MEASURES: Prevalence of obstructive CAD (any angiographic stenosis >or=50%) and incidence of adverse CV events (all-cause death or hospitalization for nonfatal myocardial infarction, stroke, congestive heart failure, unstable angina, or other vascular events) during follow-up. RESULTS: Of 906 women (mean age, 58 [SD, 12] years) with complete data, 19% were of nonwhite race, 76% were overweight (BMI >or=25), 70% had low functional capacity (DASI scores <25, equivalent to .05 for each comparison). A total of 25% of volunteers missed scheduled duties due to vaccine-related symptoms. CONCLUSIONS: Even at diluted doses, APSV is an effective smallpox vaccine, allowing for expansion of the current stockpile. However, reactogenicity was not reduced with dilution of the vaccine and, as with other smallpox vaccines, may impair daily activities. PMID- 15353534 TI - Consumer-driven health care: lessons from Switzerland. AB - Switzerland's consumer-driven health care system achieves universal insurance and high quality of care at significantly lower costs than the employer-based US system and without the constrained resources that can characterize government controlled systems. Unlike other systems in which the choice and most of the funding for health insurance is provided by third parties, such as employers and governments, in the Swiss system, individuals are required to purchase their own health insurance. The positive results achieved by the Swiss system may be attributed to its consumer control, price transparency of the insurance plans, risk adjustment of insurers, and solidarity. However, the constraints the Swiss system places on hospitals and physicians and the paucity of provider quality information may unduly limit its impact. The Swiss health care system holds important lessons, including evidence about its feasibility and equity, for the United States, which is now embarking on its own consumer-driven health care system. PMID- 15353535 TI - Near-syncope after exercise. AB - Syncope and near-syncope are great diagnostic challenges in medicine. On the one hand, the symptom may result from a benign condition and pose little or no threat to health other than that related to falling. On the other hand, syncope or near syncope can be the manifestation of a serious underlying condition that poses an imminent threat to life. Patients with a cardiac cause of syncope are at far greater risk of dying in the first year after an episode of syncope or near syncope than individuals with a noncardiac cause. A cardiac cause of syncope should be considered in every patient with syncope or near-syncope, but it is particularly common in older patients or in patients with known structural heart disease, arrhythmia, or certain electrocardiographic abnormalities. Although many diagnostic tests may be helpful in the evaluation of syncope and near-syncope, the history, physical examination, and electrocardiogram pinpoint the cause in many circumstances. Syncope after exercise may be due to left ventricular outflow tract obstruction from aortic stenosis or hypertrophic obstructive cardiomyopathy but can also suggest the diagnosis of postexercise hypotension in which an abnormality in autonomic regulation of vascular tone or heart rate results in vasodilation or bradycardia after moderate-intensity aerobic activity. The patient discussed in this case highlights the importance of the clinical history in the evaluation of this condition, since the diagnosis was revealed as the patient's story was described and eventually acted out. PMID- 15353536 TI - The Swiss health system: regulated competition without managed care. PMID- 15353537 TI - The fitness, obesity, and health equation: is physical activity the common denominator? PMID- 15353538 TI - Pharmacotherapy of chronic fatigue syndrome: another gallant attempt. PMID- 15353539 TI - JAMA patient page. Fainting. PMID- 15353540 TI - Phase II study of denileukin diftitox for relapsed/refractory B-Cell non Hodgkin's lymphoma. AB - PURPOSE: Denileukin diftitox is a fusion protein combining diphtheria toxin and interleukin-2 (IL-2) that targets tumor cells expressing the IL-2 receptor. Its efficacy has been shown in CD25+ cutaneous T-cell lymphoma, but not in B-cell non Hodgkin's lymphoma (NHL). A phase II study was performed to evaluate the efficacy and tolerability of denileukin diftitox for relapsed or refractory B-cell NHL. PATIENTS AND METHODS: Patients with relapsed or refractory B-cell NHL were eligible. Tumor CD25 expression was determined by immunohistochemistry or flow cytometry. Denileukin diftitox was administered intravenously at a dose of 18 microg/kg once daily for 5 days every 3 weeks, up to eight cycles. RESULTS: Of the 45 patients assessable for response, 32 (71%) were refractory to the last chemotherapy treatment, and all were previously treated with rituximab. Three complete responses (6.7%) and eight partial responses (17.8%) were observed, for an overall response rate of 24.5%. Nine patients (20%) had stable disease. Objective response rates were similar in CD25+ (22%) and CD25- histologies (29%), as were stable disease rates (22% and 18%, respectively). For responding patients, the median time to treatment failure was 7 months, with a median follow up in survivors of 18 months (range, 9 to 28 months), and the projected progression-free survival at 20 months was 24% (95% CI, 0% to 60%). Most toxicities were low-grade and transient. CONCLUSION: Denileukin diftitox seems to be effective in relapsed or refractory, CD25+ and CD25- B-cell NHL and is well tolerated at the dosage evaluated. Evaluation of denileukin diftitox in combination with other agents may be warranted. PMID- 15353541 TI - Cancer prevention and the American Society of Clinical Oncology. PMID- 15353542 TI - Outcome of treatment in adults with acute lymphoblastic leukemia: analysis of the LALA-94 trial. AB - PURPOSE: We analyzed the benefits of a risk-adapted postremission strategy in adult lymphoblastic leukemia (ALL), and re-evaluated stem-cell transplantation (SCT) for high-risk ALL. PATIENTS AND METHODS: A total of 922 adult patients entered onto the trial according to risk groups: standard-risk ALL (group 1), high-risk ALL (group 2), Philadelphia chromosome-positive ALL (group 3), and CNS positive ALL (group 4). All received a standard four-drug/4-week induction course. Patients from group 1 who achieved a complete remission (CR) after one course of induction therapy were randomly assigned between intensive and less intensive postremission chemotherapy, whereas those who achieved CR after salvage therapy were then included in group 2. Patients in groups 2, 3, and 4 with an HLA identical sibling were assigned to allogeneic SCT. In groups 3 and 4, autologous SCT was offered to all other patients, whereas in group 2 they were randomly assigned between chemotherapy and autologous SCT. RESULTS: Overall, 771 patients achieved CR (84%). Median disease-free survival (DFS) was 17.5 months, with 3 year DFS at 37%. In group 1, the 3-year DFS rate was 41%, with no difference between arms of postremission randomization. In groups 2 and 4, the 3-year DFS rates were 38% and 44%, respectively. In group 2, autologous SCT and chemotherapy resulted in comparable median DFS. Patients with an HLA-matched sibling (groups 2 and 4) had improved DFS. Three-year DFS was 24% in group 3. CONCLUSION: Allogeneic SCT improved DFS in high-risk ALL in the first CR. Autologous SCT did not confer a significant benefit over chemotherapy for high-risk ALL. PMID- 15353543 TI - Autologous stem-cell transplantation can be performed safely without the use of blood-product support. AB - PURPOSE: Autologous stem-cell transplantation has been shown to be a curative procedure for a variety of leukemias and lymphomas. Most transplants require RBC and platelet support. We report the ability to perform autologous transplantation without blood-product support. SUBJECTS AND METHODS: In this study, we treated 26 patients with religious objection to blood products with autologous stem-cell support without the use of any blood products. Patients received a combination of granulocyte colony-stimulating factor (G-CSF), erythropoietin, and interleukin-11 or G-CSF alone to mobilize stem cells. Post-transplant patients received intravenous iron, erythropoietin, G-CSF, and epsilon aminocaproic acid. RESULTS: There were two major bleeding complications (8%), with two treatment-related deaths (8%). There were three minor bleeding complications (12%). The median fall in hemoglobin level was 4.7 g/dL; the median hemoglobin level 30 days after transplantation was 9.2 g/dL. The median total number of days with platelet count less than 10 x 10(9)/L was 4 days; the median days to platelet recovery greater than 20 x 10(9)/L was 12 days. CONCLUSION: Autologous stem-cell transplantation can be performed safely without the use of any blood products. PMID- 15353544 TI - Impacts of a new transcription factor family: mammalian GCM proteins in health and disease. AB - GCM proteins constitute a small transcription factor family with a DNA-binding domain exhibiting a novel fold composed of two subdomains rigidly held together by coordination of one of two structural zinc cations. In all known cases, GCM proteins exert the role of master regulators: the prototypical family member determines gliogenesis in Drosophila melanogaster, whereas mammalian GCM proteins orchestrate divergent aspects of development and physiology in placenta, kidney, thymus, and parathyroid gland. Recent data point to an involvement of GCM proteins in different pathological contexts, such as preeclampsia, hyper- or hypoparathyroidism, and parathyroid gland tumors. PMID- 15353545 TI - Visualization of early chromosome condensation: a hierarchical folding, axial glue model of chromosome structure. AB - Current models of mitotic chromosome structure are based largely on the examination of maximally condensed metaphase chromosomes. Here, we test these models by correlating the distribution of two scaffold components with the appearance of prophase chromosome folding intermediates. We confirm an axial distribution of topoisomerase IIalpha and the condensin subunit, structural maintenance of chromosomes 2 (SMC2), in unextracted metaphase chromosomes, with SMC2 localizing to a 150-200-nm-diameter central core. In contrast to predictions of radial loop/scaffold models, this axial distribution does not appear until late prophase, after formation of uniformly condensed middle prophase chromosomes. Instead, SMC2 associates throughout early and middle prophase chromatids, frequently forming foci over the chromosome exterior. Early prophase condensation occurs through folding of large-scale chromatin fibers into condensed masses. These resolve into linear, 200-300-nm-diameter middle prophase chromatids that double in diameter by late prophase. We propose a unified model of chromosome structure in which hierarchical levels of chromatin folding are stabilized late in mitosis by an axial "glue." PMID- 15353546 TI - Robust cell polarity is a dynamic state established by coupling transport and GTPase signaling. AB - Yeast cells can initiate bud formation at the G1/S transition in a cue independent manner. Here, we investigate the dynamic nature of the polar cap and the regulation of the GTPase Cdc42 in the establishment of cell polarity. Using analysis of fluorescence recovery after photobleaching, we found that Cdc42 exchanged rapidly between the polar caps and cytosol and that this rapid exchange required its GTPase cycle. A previously proposed positive feedback loop involving actomyosin-based transport of the Cdc42 GTPase is required for the generation of robust cell polarity during bud formation in yeast. Inhibition of actin-based transport resulted in unstable Cdc42 polar caps. Unstable polarity was also observed in mutants lacking Bem1, a protein previously implicated in a feedback loop for Cdc42 activation through a signaling pathway. When Bem1 and actin were both inhibited, polarization completely failed. These results suggest that cell polarity is established through coupling of transport and signaling pathways and maintained actively by balance of flux. PMID- 15353547 TI - Quantitative kinetic analysis of nucleolar breakdown and reassembly during mitosis in live human cells. AB - One of the great mysteries of the nucleolus surrounds its disappearance during mitosis and subsequent reassembly at late mitosis. Here, the relative dynamics of nucleolar disassembly and reformation were dissected using quantitative 4D microscopy with fluorescent protein-tagged proteins in human stable cell lines. The data provide a novel insight into the fates of the three distinct nucleolar subcompartments and their associated protein machineries in a single dividing cell. Before the onset of nuclear envelope (NE) breakdown, nucleolar disassembly started with the loss of RNA polymerase I subunits from the fibrillar centers. Dissociation of proteins from the other subcompartments occurred with faster kinetics but commenced later, coincident with the process of NE breakdown. The reformation pathway also follows a reproducible and defined temporal sequence but the order of reassembly is shown not to be dictated by the order in which individual nucleolar components reaccumulate within the nucleus after mitosis. PMID- 15353548 TI - Phosphorylation of actopaxin regulates cell spreading and migration. AB - Actopaxin is an actin and paxillin binding protein that localizes to focal adhesions. It regulates cell spreading and is phosphorylated during mitosis. Herein, we identify a role for actopaxin phosphorylation in cell spreading and migration. Stable clones of U2OS cells expressing actopaxin wild-type (WT), nonphosphorylatable, and phosphomimetic mutants were developed to evaluate actopaxin function. All proteins targeted to focal adhesions, however the nonphosphorylatable mutant inhibited spreading whereas the phosphomimetic mutant cells spread more efficiently than WT cells. Endogenous and WT actopaxin, but not the nonphosphorylatable mutant, were phosphorylated in vivo during cell adhesion/spreading. Expression of the nonphosphorylatable actopaxin mutant significantly reduced cell migration, whereas expression of the phosphomimetic increased cell migration in scrape wound and Boyden chamber migration assays. In vitro kinase assays demonstrate that extracellular signal-regulated protein kinase phosphorylates actopaxin, and treatment of U2OS cells with the MEK1 inhibitor UO126 inhibited adhesion-induced phosphorylation of actopaxin and also inhibited cell migration. PMID- 15353549 TI - p107 regulates neural precursor cells in the mammalian brain. AB - Here we show a novel function for Retinoblastoma family member, p107 in controlling stem cell expansion in the mammalian brain. Adult p107-null mice had elevated numbers of proliferating progenitor cells in their lateral ventricles. In vitro neurosphere assays revealed striking increases in the number of neurosphere forming cells from p107(-/-) brains that exhibited enhanced capacity for self-renewal. An expanded stem cell population in p107-deficient mice was shown in vivo by (a) increased numbers of slowly cycling cells in the lateral ventricles; and (b) accelerated rates of neural precursor repopulation after progenitor ablation. Notch1 was up-regulated in p107(-/-) neurospheres in vitro and brains in vivo. Chromatin immunoprecipitation and p107 overexpression suggest that p107 may modulate the Notch1 pathway. These results demonstrate a novel function for p107 that is distinct from Rb, which is to negatively regulate the number of neural stem cells in the developing and adult brain. PMID- 15353550 TI - A novel checkpoint in the Bcl-2-regulated apoptotic pathway revealed by murine cytomegalovirus infection of dendritic cells. AB - Infection with murine cytomegalovirus (MCMV) has contributed to understanding many aspects of human infection and, additionally, has provided important insight to understanding complex cellular responses. Dendritic cells (DCs) are a major target for MCMV infection. Here, we analyze the effects of MCMV infection on DC viability, and show that infected DCs become resistant to apoptosis induced by growth factor deprivation. The precise contribution of changes in the expression of Bcl-2 family proteins has been assessed and a new checkpoint in the apoptotic pathway identified. Despite their resistance to apoptosis, MCMV-infected DCs showed Bax to be tightly associated with mitochondria and, together with Bak, forming high molecular weight oligomers, changes normally associated with apoptotic cell death. Exposure of a constitutively occluded Bax NH2-terminal epitope was blocked after infection. These results suggest that MCMV has evolved a novel strategy for inhibiting apoptosis and provide evidence that apoptosis can be regulated after translocation, integration, and oligomerization of Bax at the mitochondrial membrane. PMID- 15353551 TI - The PYRIN connection: novel players in innate immunity and inflammation. AB - Periodic fever syndromes (PFSs) comprise a subset of the hereditary autoinflammatory disorders that are defined by recurrent self-resolving attacks of systemic inflammatory reactions in the absence of infection or autoimmunity. Recent advances have led to the discovery that members of a new family of genes, the PYRIN family, account for several hereditary PFSs. Here we discuss new insights into the function of PYRIN proteins and the molecular basis of PFSs. PMID- 15353552 TI - Decreased survival of B cells of HIV-viremic patients mediated by altered expression of receptors of the TNF superfamily. AB - Human immunodeficiency virus (HIV) infection leads to numerous perturbations of B cells through mechanisms that remain elusive. We performed DNA microarray, phenotypic, and functional analyses in an effort to elucidate mechanisms of B cell perturbation associated with ongoing HIV replication. 42 genes were up regulated in B cells of HIV-viremic patients when compared with HIV-aviremic and HIV-negative patients, the majority of which were interferon (IFN)-stimulated or associated with terminal differentiation. Flow cytometry confirmed these increases and indicated that CD21(low) B cells, enhanced in HIV-viremic patients, were largely responsible for the changes. Increased expression of the tumor necrosis factor (TNF) superfamily (TNFSF) receptor CD95 correlated with increased susceptibility to CD95-mediated apoptosis of CD21(low) B cells, which, in turn, correlated with HIV plasma viremia. Increased expression of BCMA, a weak TNFSF receptor for B lymphocyte stimulator (BLyS), on CD21(low) B cells was associated with a concomitant reduction in the expression of the more potent BLyS receptor, BAFF-R, that resulted in reduced BLyS binding and BLyS-mediated survival. These findings demonstrate that altered expression of genes associated with IFN stimulation and terminal differentiation in B cells of HIV-viremic patients lead to an increased propensity to cell death, which may have substantial deleterious effects on B cell responsiveness to antigenic stimulation. PMID- 15353554 TI - Determination of the critical concentration of neutrophils required to block bacterial growth in tissues. AB - We showed previously that the competition between bacterial killing by neutrophils and bacterial growth in stirred serum-containing suspensions could be modeled as the competition between a first-order reaction (bacterial growth) and a second-order reaction (bacterial killing by neutrophils). The model provided a useful parameter, the critical neutrophil concentration (CNC), below which bacterial concentration increased and above which it decreased, independent of the initial bacterial concentration. We report here that this model applies to neutrophil killing of bacteria in three-dimensional fibrin matrices and in rabbit dermis. We measured killing of 10(3)-10(8) colony forming units/ml Staphylococcus epidermidis by 10(5)-10(8) human neutrophils/ml in fibrin gels. The CNC was approximately 4 x 10(6) neutrophils/ml gel in the presence of normal serum and approximately 1.6 x 10(7) neutrophils/ml gel in the presence of C5-deficient serum. Application of our model to published data of others on killing of approximately 5 x 10(7) to 2 x 10(8) E. coli/ml rabbit dermis yielded CNCs from approximately 4 x 10(6) to approximately 8 x 10(6) neutrophils/ml dermis. Thus, in disparate tissues and tissuelike environments, our model fits the kinetics of bacterial killing and gives similar lower limits (CNCs) to the neutrophil concentration required to control bacterial growth. PMID- 15353553 TI - MyD88 but not TRIF is essential for osteoclastogenesis induced by lipopolysaccharide, diacyl lipopeptide, and IL-1alpha. AB - Myeloid differentiation factor 88 (MyD88) plays essential roles in the signaling of the Toll/interleukin (IL)-1 receptor family. Toll-IL-1 receptor domain containing adaptor inducing interferon-beta (TRIF)-mediated signals are involved in lipopolysaccharide (LPS)-induced MyD88-independent pathways. Using MyD88 deficient (MyD88-/-) mice and TRIF-deficient (TRIF-/-) mice, we examined roles of MyD88 and TRIF in osteoclast differentiation and function. LPS, diacyl lipopeptide, and IL-1alpha stimulated osteoclastogenesis in cocultures of osteoblasts and hemopoietic cells obtained from TRIF-/- mice, but not MyD88-/- mice. These factors stimulated receptor activator of nuclear factor-kappaB ligand mRNA expression in TRIF-/- osteoblasts, but not MyD88-/- osteoblasts. LPS stimulated IL-6 production in TRIF-/- osteoblasts, but not TRIF-/- macrophages. LPS and IL-1alpha enhanced the survival of TRIF-/- osteoclasts, but not MyD88-/- osteoclasts. Diacyl lipopeptide did not support the survival of osteoclasts because of the lack of Toll-like receptor (TLR)6 in osteoclasts. Macrophages expressed both TRIF and TRIF-related adaptor molecule (TRAM) mRNA, whereas osteoblasts and osteoclasts expressed only TRIF mRNA. Bone histomorphometry showed that MyD88-/- mice exhibited osteopenia with reduced bone resorption and formation. These results suggest that the MyD88-mediated signal is essential for the osteoclastogenesis and function induced by IL-1 and TLR ligands, and that MyD88 is physiologically involved in bone turnover. PMID- 15353555 TI - Constitutive activation of STAT5A promotes human hematopoietic stem cell self renewal and erythroid differentiation. AB - Activation of the transcription factor signal transducer and activator of transcription (STAT)5 is involved in various aspects of hematopoiesis, affecting cell proliferation, differentiation, and cell survival. Constitutive activation of STAT5 has also been associated with leukemic transformation. We overexpressed the constitutively active mutant STAT5A(1*6) in human cord blood CD34+ cells and evaluated the effects on the hematopoietic potential of stem cells in a variety of in vitro and in vivo systems. The observed phenotypic changes were correlated with differential gene expression patterns induced by STAT5A(1*6). Our data indicate that a persistent activation of STAT5A in human hematopoietic stem and progenitor cells results in their enhanced self-renewal and diverts differentiation to the erythroid lineage. PMID- 15353556 TI - Molecular characterization of virus-induced autoantibody responses. AB - Here we present a comprehensive molecular mapping of virus-induced autoimmune B cell responses obtained by serological identification of antigens by recombinant expression cloning analysis. Immunoscreening of cDNA expression libraries of various organs (lung, liver, and spleen) using sera from mice infected with cytopathic (vaccinia virus [VV]) or noncytopathic (lymphocytic choriomeningitis virus [LCMV]) viruses revealed a broad specificity of the elicited autoantibody response. Interestingly, the majority of the identified autoantigens have been previously described as autoantigens in humans. We found that induction of virus induced autoantibodies of the immunoglobulin G class largely depends on the CD40 CD40L-mediated interaction between T and B cells. Furthermore, antibody titers against a number of autoantigens were comparable to the concomitantly induced antiviral antibody response. Comparison of serum reactivity against a selected panel of autoantigens after infection with VV, LCMV, or vesicular stomatitis virus showed that the different virus infections triggered distinct autoantibody responses, suggesting that virus infections may leave specific "autoantibody fingerprints" in the infected host. PMID- 15353557 TI - Dormancy phenotype displayed by extracellular Mycobacterium tuberculosis within artificial granulomas in mice. AB - Mycobacterium tuberculosis residing within pulmonary granulomas and cavities represents an important reservoir of persistent organisms during human latent tuberculosis infection. We present a novel in vivo model of tuberculosis involving the encapsulation of bacilli in semidiffusible hollow fibers that are implanted subcutaneously into mice. Granulomatous lesions develop around these hollow fibers, and in this microenvironment, the organisms demonstrate an altered physiologic state characterized by stationary-state colony-forming unit counts and decreased metabolic activity. Moreover, these organisms show an antimicrobial susceptibility pattern similar to persistent bacilli in current models of tuberculosis chemotherapy in that they are more susceptible to the sterilizing drug, rifampin, than to the bactericidal drug isoniazid. We used this model of extracellular persistence within host granulomas to study both gene expression patterns and mutant survival patterns. Our results demonstrate induction of dosR (Rv3133c) and 20 other members of the DosR regulon believed to mediate the transition into dormancy, and that rel(Mtb) is required for Mycobacterium tuberculosis survival during extracellular persistence within host granulomas. Interestingly, the dormancy phenotype of extracellular M. tuberculosis within host granulomas appears to be immune mediated and interferon-gamma dependent. PMID- 15353558 TI - Activation of PI3K is indispensable for interleukin 7-mediated viability, proliferation, glucose use, and growth of T cell acute lymphoblastic leukemia cells. AB - Interleukin (IL)-7 is essential for normal T cell development. Previously, we have shown that IL-7 increases viability and proliferation of T cell acute lymphoblastic leukemia (T-ALL) cells by up-regulating Bcl-2 and down-regulating the cyclin-dependent kinase inhibitor p27kip1. Here, we examined the signaling pathways via which IL-7 mediates these effects. We investigated mitogen-activated protein kinase (MEK)-extracellular signal-regulated kinase (Erk) and phosphatidylinositol-3-kinase (PI3K)-Akt (protein kinase B) pathways, which have active roles in T cell expansion and have been implicated in tumorigenesis. IL-7 induced activation of the MEK-Erk pathway in T-ALL cells; however, inhibition of the MEK-Erk pathway by the use of the cell-permeable inhibitor PD98059, did not affect IL-7-mediated viability or cell cycle progression of leukemic cells. IL-7 induced PI3K-dependent phosphorylation of Akt and its downstream targets GSK-3, FOXO1, and FOXO3a. PI3K activation was mandatory for IL-7-mediated Bcl-2 up regulation, p27kip1 down-regulation, Rb hyperphosphorylation, and consequent viability and cell cycle progression of T-ALL cells. PI3K signaling was also required for cell size increase, up-regulation of CD71, expression of the glucose transporter Glut1, uptake of glucose, and maintenance of mitochondrial integrity. Our results implicate PI3K as a major effector of IL-7-induced viability, metabolic activation, growth and proliferation of T-ALL cells, and suggest that PI3K and its downstream effectors may represent molecular targets for therapeutic intervention in T-ALL. PMID- 15353559 TI - NMR studies of the structure and Mg2+ binding properties of a conserved RNA motif of EMCV picornavirus IRES element. AB - The structure and Mg(2+) binding properties of a conserved 75mer RNA motif of the internal ribosome entry site (IRES) element of encephalomyocarditis virus picornavirus have been investigated by (1)H-NMR and UV melting experiments. The assignment of the imino proton resonances with characteristic chemical shift dispersion for canonical and non-canonical base pairs confirmed the predicted secondary structure of the 75mer and its fragments. Addition of Mg(2+) resulted in a dramatic increase in apparent melting temperature, with the 75mer RNA registering the biggest increase, from 63 to 80 degrees C, thus providing evidence for enhanced stability arising from Mg(2+) binding. Similarly, addition of Mg(2+) induced selective changes to the chemical shifts of the imino protons of a GCGA tetraloop in the 75mer, that is essential for IRES activity, thereby highlighting a possible structural role for Mg(2+) in the folding of the 75mer. Significantly, the same protons show retarded exchange to water solvent, even at elevated temperature, which suggest that Mg(2+) induces a conformational rearrangement of the 75mer. Thus, we propose that Mg(2+) serves two important roles: (i) enhancing thermodynamic stability of the 75mer RNA (and its submotifs) via non-specific interactions with the phosphate backbone and (ii) promoting the folding of the 75mer RNA by binding to the GCGA tetraloop. PMID- 15353560 TI - Gene co-regulation is highly conserved in the evolution of eukaryotes and prokaryotes. AB - Differences between species have been suggested to largely reside in the network of connections among the genes. Nevertheless, the rate at which these connections evolve has not been properly quantified. Here, we measure the extent to which co regulation between pairs of genes is conserved over large phylogenetic distances; between two eukaryotes Caenorhabditis elegans and Saccharomyces cerevisiae, and between two prokaryotes Escherichia coli and Bacillus subtilis. We first construct a reliable set of co-regulated genes by combining various functional genomics data from yeast, and subsequently determine conservation of co regulation in worm from the distribution of co-expression values. For B.subtilis and E.coli, we use known operons and regulons. We find that between 76 and 80% of the co-regulatory connections are conserved between orthologous pairs of genes, which is very high compared with previous estimates and expectations regarding network evolution. We show that in the case of gene duplication after speciation, one of the two inparalogous genes tends to retain its original co-regulatory relationship, while the other loses this link and is presumably free for differentiation or sub-functionalization. The high level of co-regulation conservation implies that reliably predicted functional relationships from functional genomics data in one species can be transferred with high accuracy to another species when that species also harbours the associated genes. PMID- 15353561 TI - A novel real-time PCR assay for quantitative analysis of methylated alleles (QAMA): analysis of the retinoblastoma locus. AB - Altered methylation patterns have been found to play a role in developmental disorders, cancer and aging. Increasingly, changes in DNA methylation are used as molecular markers of disease. Therefore, there is a need for reliable and easy to use techniques to detect and measure DNA methylation in research and routine diagnostics. We have established a novel quantitative analysis of methylated alleles (QAMA) which is essentially a major improvement over a previous method based on real-time PCR (MethyLight). This method is based on real-time PCR on bisulfite-treated DNA. A significant advantage over conventional MethyLight is gained by the use of TaqMan probes based on minor groove binder (MGB) technology. Their improved sequence specificity facilitates relative quantification of methylated and unmethylated alleles that are simultaneously amplified in single tube. This improvement allows precise measurement of the ratio of methylated versus unmethylated alleles and cuts down potential sources of inter-assay variation. Therefore, fewer control assays are required. We have used this novel technical approach to identify hypermethylation of the CpG island located in the promoter region of the retinoblastoma (RB1) gene and found that QAMA facilitates reliable and fast measurement of the relative quantity of methylated alleles and improves handling of diagnostic methylation analysis. Moreover, the simplified reaction setup and robustness inherent to the single tube assay facilitates high throughput methylation analysis. Because the high sequence specificity inherent to the MGB technology is widely used to discriminate single nucleotide polymorphisms, QAMA potentially can be used to discriminate the methylation status of single CpG dinucleotides. PMID- 15353563 TI - Biodiversity of vibrios. AB - Vibrios are ubiquitous and abundant in the aquatic environment. A high abundance of vibrios is also detected in tissues and/or organs of various marine algae and animals, e.g., abalones, bivalves, corals, fish, shrimp, sponges, squid, and zooplankton. Vibrios harbour a wealth of diverse genomes as revealed by different genomic techniques including amplified fragment length polymorphism, multilocus sequence typing, repetetive extragenic palindrome PCR, ribotyping, and whole genome sequencing. The 74 species of this group are distributed among four different families, i.e., Enterovibrionaceae, Photobacteriaceae, Salinivibrionaceae, and Vibrionaceae. Two new genera, i.e., Enterovibrio norvegicus and Grimontia hollisae, and 20 novel species, i.e., Enterovibrio coralii, Photobacterium eurosenbergii, V. brasiliensis, V. chagasii, V. coralliillyticus, V. crassostreae, V. fortis, V. gallicus, V. hepatarius, V. hispanicus, V. kanaloaei, V. neonatus, V. neptunius, V. pomeroyi, V. pacinii, V. rotiferianus, V. superstes, V. tasmaniensis, V. ezurae, and V. xuii, have been described in the last few years. Comparative genome analyses have already revealed a variety of genomic events, including mutations, chromosomal rearrangements, loss of genes by decay or deletion, and gene acquisitions through duplication or horizontal transfer (e.g., in the acquisition of bacteriophages, pathogenicity islands, and super-integrons), that are probably important driving forces in the evolution and speciation of vibrios. Whole-genome sequencing and comparative genomics through the application of, e.g., microarrays will facilitate the investigation of the gene repertoire at the species level. Based on such new genomic information, the taxonomy and the species concept for vibrios will be reviewed in the next years. PMID- 15353562 TI - Binary bacterial toxins: biochemistry, biology, and applications of common Clostridium and Bacillus proteins. AB - Certain pathogenic species of Bacillus and Clostridium have developed unique methods for intoxicating cells that employ the classic enzymatic "A-B" paradigm for protein toxins. The binary toxins produced by B. anthracis, B. cereus, C. botulinum, C. difficile, C. perfringens, and C. spiroforme consist of components not physically associated in solution that are linked to various diseases in humans, animals, or insects. The "B" components are synthesized as precursors that are subsequently activated by serine-type proteases on the targeted cell surface and/or in solution. Following release of a 20-kDa N-terminal peptide, the activated "B" components form homoheptameric rings that subsequently dock with an "A" component(s) on the cell surface. By following an acidified endosomal route and translocation into the cytosol, "A" molecules disable a cell (and host organism) via disruption of the actin cytoskeleton, increasing intracellular levels of cyclic AMP, or inactivation of signaling pathways linked to mitogen activated protein kinase kinases. Recently, B. anthracis has gleaned much notoriety as a biowarfare/bioterrorism agent, and of primary interest has been the edema and lethal toxins, their role in anthrax, as well as the development of efficacious vaccines and therapeutics targeting these virulence factors and ultimately B. anthracis. This review comprehensively surveys the literature and discusses the similarities, as well as distinct differences, between each Clostridium and Bacillus binary toxin in terms of their biochemistry, biology, genetics, structure, and applications in science and medicine. The information may foster future studies that aid novel vaccine and drug development, as well as a better understanding of a conserved intoxication process utilized by various gram-positive, spore-forming bacteria. PMID- 15353565 TI - CO-sensing mechanisms. AB - Carbon monoxide (CO) has long been known to have dramatic physiological effects on organisms ranging from bacteria to humans, but recently there have a number of suggestions that organisms might have specific sensors for CO. This article reviews the current evidence for a variety of proteins with demonstrated or potential CO-sensing ability. Particular emphasis is placed on the molecular description of CooA, a heme-containing CO sensor from Rhodospirillum rubrum, since its biological role as a CO sensor is clear and we have substantial insight into the basis of its sensing ability. PMID- 15353564 TI - Effects of length and location on the cellular response to double-stranded RNA. AB - Since double-stranded RNA (dsRNA) has not until recently generally been thought to be deliberately expressed in cells, it has commonly been assumed that the major source of cellular dsRNA is viral infections. In this view, the cellular responses to dsRNA would be natural and perhaps ancient antiviral responses. While the cell may certainly react to some dsRNAs as an antiviral response, this does not represent the only response or even, perhaps, the major one. A number of recent observations have pointed to the possibility that dsRNA molecules are not seen only as evidence of viral infection or recognized for degradation because they cannot be translated. In some instances they may also play important roles in normal cell growth and function. The purpose of this review is to outline our current understanding of the fate of dsRNA in cells, with a focus on the apparent fact that their fates and functions appear to depend critically not only on where in the cell dsRNA molecules are found, but also on how long they are and perhaps on how abundant they are. PMID- 15353566 TI - Bacterial transcriptional regulators for degradation pathways of aromatic compounds. AB - Human activities have resulted in the release and introduction into the environment of a plethora of aromatic chemicals. The interest in discovering how bacteria are dealing with hazardous environmental pollutants has driven a large research community and has resulted in important biochemical, genetic, and physiological knowledge about the degradation capacities of microorganisms and their application in bioremediation, green chemistry, or production of pharmacy synthons. In addition, regulation of catabolic pathway expression has attracted the interest of numerous different groups, and several catabolic pathway regulators have been exemplary for understanding transcription control mechanisms. More recently, information about regulatory systems has been used to construct whole-cell living bioreporters that are used to measure the quality of the aqueous, soil, and air environment. The topic of biodegradation is relatively coherent, and this review presents a coherent overview of the regulatory systems involved in the transcriptional control of catabolic pathways. This review summarizes the different regulatory systems involved in biodegradation pathways of aromatic compounds linking them to other known protein families. Specific attention has been paid to describing the genetic organization of the regulatory genes, promoters, and target operon(s) and to discussing present knowledge about signaling molecules, DNA binding properties, and operator characteristics, and evidence from regulatory mutants. For each regulator family, this information is combined with recently obtained protein structural information to arrive at a possible mechanism of transcription activation. This demonstrates the diversity of control mechanisms existing in catabolic pathways. PMID- 15353567 TI - Microbial type I fatty acid synthases (FAS): major players in a network of cellular FAS systems. AB - The present review focuses on microbial type I fatty acid synthases (FASs), demonstrating their structural and functional diversity. Depending on their origin and biochemical function, multifunctional type I FAS proteins form dimers or hexamers with characteristic organization of their catalytic domains. A single polypeptide may contain one or more sets of the eight FAS component functions. Alternatively, these functions may split up into two different and mutually complementing subunits. Targeted inactivation of the individual yeast FAS acylation sites allowed us to define their roles during the overall catalytic process. In particular, their pronounced negative cooperativity is presumed to coordinate the FAS initiation and chain elongation reactions. Expression of the unlinked genes, FAS1 and FAS2, is in part constitutive and in part subject to repression by the phospholipid precursors inositol and choline. The interplay of the involved regulatory proteins, Rap1, Reb1, Abf1, Ino2/Ino4, Opi1, Sin3 and TFIIB, has been elucidated in considerable detail. Balanced levels of subunits alpha and beta are ensured by an autoregulatory effect of FAS1 on FAS2 expression and by posttranslational degradation of excess FAS subunits. The functional specificity of type I FAS multienzymes usually requires the presence of multiple FAS systems within the same cell. De novo synthesis of long-chain fatty acids, mitochondrial fatty acid synthesis, acylation of certain secondary metabolites and coenzymes, fatty acid elongation, and the vast diversity of mycobacterial lipids each result from specific FAS activities. The microcompartmentalization of FAS activities in type I multienzymes may thus allow for both the controlled and concerted action of multiple FAS systems within the same cell. PMID- 15353568 TI - Determination of the core of a minimal bacterial gene set. AB - The availability of a large number of complete genome sequences raises the question of how many genes are essential for cellular life. Trying to reconstruct the core of the protein-coding gene set for a hypothetical minimal bacterial cell, we have performed a computational comparative analysis of eight bacterial genomes. Six of the analyzed genomes are very small due to a dramatic genome size reduction process, while the other two, corresponding to free-living relatives, are larger. The available data from several systematic experimental approaches to define all the essential genes in some completely sequenced bacterial genomes were also considered, and a reconstruction of a minimal metabolic machinery necessary to sustain life was carried out. The proposed minimal genome contains 206 protein-coding genes with all the genetic information necessary for self maintenance and reproduction in the presence of a full complement of essential nutrients and in the absence of environmental stress. The main features of such a minimal gene set, as well as the metabolic functions that must be present in the hypothetical minimal cell, are discussed. PMID- 15353571 TI - The renal protective effects of angiotensin II receptor blockers in type 2 diabetes mellitus. AB - OBJECTIVE: To review the renal protective effects of angiotensin II receptor blockers (ARBs) in patients with type 2 diabetes mellitus. DATA SOURCES: A MEDLINE search (1966-March 2004) was completed using irbesartan, candesartan, losartan, valsartan, eprosartan, olmesartan, telmisartan, renal protection, nephropathy, albuminuria, and type 2 diabetes mellitus as key words. STUDY SELECTION AND DATA EXTRACTION: All identified English-language articles were reviewed. References of the identified sources were used to identify additional articles. Articles representative of the subject matter of our review were included. DATA SYNTHESIS: ARBs have extensive data showing their renal protective benefits in hypertensive type 2 diabetic patients with microalbuminuria or proteinuria. The benefits are over and above that of blood pressure reduction alone and extend to normotensive diabetic patients as well. Maximizing the ARB dose before adding additional therapies or another renal-protecting agent (angiotensin-converting enzyme [ACE] inhibitor or non-dihydropyridine calcium channel blocker) may be superior to adding another class of antihypertensive, even if similar blood pressures can be achieved. CONCLUSIONS: ARBs are an important therapy for hypertensive type 2 diabetic patients and can benefit normotensive diabetic patients as well. ARB dosage optimization or the addition of a second renoprotective agent (ACE inhibitor or non-dihydropyridine calcium channel blocker) may be important for optimal renoprotection, although further research is clearly needed in this area. PMID- 15353575 TI - Atazanavir. AB - OBJECTIVE: To review the pharmacology, virology, pharmacokinetics, resistance profile, clinical efficacy, safety, and drug interactions of atazanavir. DATA SOURCES: A PubMed and NLMGateway search (1966-June 2004) utilizing the key words atazanavir and BMS-232632 was performed. Abstracts from scientific meetings, including the Conference on Retroviruses and Opportunistic Infections, International AIDS Society Conference on HIV Pathogenesis and Treatment, Interscience Conference on Antimicrobial Agents and Chemotherapy, and the Infectious Diseases Society of America, were also reviewed. STUDY SELECTION AND DATA EXTRACTION: All publications and meeting abstracts were reviewed, and information relevant to the formulary decision-making process was selected. DATA SYNTHESIS: Atazanavir is a once-daily protease inhibitor (PI) that received approval by the Food and Drug Administration in June 2003. In clinical trials in antiretroviral (ARV)-naive patients, atazanavir had efficacy similar to that of efavirenz or nelfinavir. In ARV-experienced patients, atazanavir was inferior to lopinavir/ritonavir unless atazanavir was coadministered with low-dose ritonavir. Following failure of an atazanavir-containing regimen in ARV-naive patients, a unique 150L mutation was seen. Atazanavir resistance is likely when resistance to >/=3 PIs is present. Atazanavir can cause increases in unconjugated bilirubin levels, which rarely leads to jaundice or scleral icterus. In contrast to comparators, atazanavir did not negatively impact the lipid profile. Similar to other PIs, atazanavir is metabolized by and inhibits CYP3A at clinically relevant concentrations; therefore, many potential drug interactions exist. CONCLUSIONS: Atazanavir is a once-daily PI that, unlike other PIs, does not negatively impact the lipid profile. Atazanavir may be particularly desirable in patients with hyperlipidemia or other coronary artery disease risk factors. PMID- 15353569 TI - Single-cell microbiology: tools, technologies, and applications. AB - The field of microbiology has traditionally been concerned with and focused on studies at the population level. Information on how cells respond to their environment, interact with each other, or undergo complex processes such as cellular differentiation or gene expression has been obtained mostly by inference from population-level data. Individual microorganisms, even those in supposedly "clonal" populations, may differ widely from each other in terms of their genetic composition, physiology, biochemistry, or behavior. This genetic and phenotypic heterogeneity has important practical consequences for a number of human interests, including antibiotic or biocide resistance, the productivity and stability of industrial fermentations, the efficacy of food preservatives, and the potential of pathogens to cause disease. New appreciation of the importance of cellular heterogeneity, coupled with recent advances in technology, has driven the development of new tools and techniques for the study of individual microbial cells. Because observations made at the single-cell level are not subject to the "averaging" effects characteristic of bulk-phase, population-level methods, they offer the unique capacity to observe discrete microbiological phenomena unavailable using traditional approaches. As a result, scientists have been able to characterize microorganisms, their activities, and their interactions at unprecedented levels of detail. PMID- 15353576 TI - Evaluation of lamotrigine toxicity reported to poison centers. AB - BACKGROUND: Lamotrigine is an antiepileptic drug for the treatment of partial and generalized seizures as well as bipolar disorder. Limited published information exists describing the clinical effects of lamotrigine overdose. OBJECTIVE: To examine the toxicity of exposures to lamotrigine utilizing national poison center data. METHODS: Data on single-substance exposures to lamotrigine reported to the American Association of Poison Control Centers Toxic Exposure Surveillance System in 2000 and 2001 were retrospectively analyzed. RESULTS: There were 493 cases that met the inclusion criteria. The majority of exposures occurred within the age groups 20-59 years old (n = 198, 40.2%), followed by 2 years. METHODS: We calculated the slopes of serum creatinine(-1)/year and urinary protein excretion/year to follow graft function over time. Subjects were genotyped for the deletion (D) polymorphism of the gene encoding angiotensin I-converting enzyme, the angiotensin II-receptor type1 gene 1166A-C polymorphism and the M235T polymorphism of the angiotensinogen gene. RESULTS: The frequencies of factors predicting graft function were similar in patients with different genotypes. None of the polymorphisms influenced need for dialysis in the first week after transplantation, occurrence of at least one rejection episode, the slope of serum creatinine(-1)/year or the slope of urinary protein excretion/year. Results were independent of blood pressure or the use of angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers or calcineurin inhibitors. The combination of genotypes did not influence the indicators of early and long-term graft dysfunction. CONCLUSIONS: Neither the investigated gene polymorphisms of the RAS in kidney allograft recipients nor their combinations have an impact on early and long-term graft dysfunction. PMID- 15353578 TI - Urinary monocyte chemoattractant protein-1 (MCP-1) is a marker of active renal vasculitis. AB - BACKGROUND: Macrophage infiltration and cytokine production are important in the pathogenesis of crescentic glomerulonephritis in anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitis. The aim of this study was to investigate whether urinary levels of chemokines, monocyte chemoattractant protein-1 (MCP-1) and fractalkine, were useful tools for non-invasive assessment of renal vasculitis. METHODS: In a prospective study, concentrations of chemokines were measured in urine and serum samples using specific enzyme-linked immunosorbent assays, and related to the patients' clinical status. Renal expression of MCP-1 was studied by immunohistochemical staining of renal biopsies. RESULTS: Urinary levels of MCP-1 were significantly higher in patients with active (P<0.01) or persistent (P<0.05) renal vasculitis, in comparison with healthy volunteers, control patients, patients with inactive vasculitis and patients with extra-renal disease only. There were no differences in serum concentrations of MCP-1 between these groups. Reduction in urinary MCP-1 levels following treatment preceded the improvement of renal function by a median of 2 weeks. In one patient, rising urinary levels of MCP-1, despite immunosuppressive therapy, was associated with progression to severe renal failure. There were no differences in urinary fractalkine levels between the different groups of patients and controls. Immunohistology of renal biopsies from patients with crescentic glomerulonephritis showed increased staining for MCP-1 in glomerular and interstitial cells. Urinary MCP-1 levels correlated with glomerular, but not tubulointerstitial, macrophage infiltration (P<0.05). CONCLUSIONS: This study shows that measurement of urinary MCP-1, but not fractalkine, is a useful non invasive technique for the assessment of renal involvement and monitoring the response to therapy in ANCA-associated vasculitis. PMID- 15353579 TI - Suppressed T-cell activation by IFN-gamma-induced expression of PD-L1 on renal tubular epithelial cells. AB - BACKGROUND: The interaction of the T-cell molecule PD-1 (programmed death-1) with its ligands PD-L1 and PD-L2 represents a known mechanism of T-cell inhibition. PD 1 is homologous to CD28 while the PD-1 ligands share homology with the B7 family of co-stimulatory molecules. METHODS: We have studied surface expression and transcript levels of PD-L1 and PD-L2 on murine renal tubular epithelial cells (TEC) by flow cytometric analysis and reverse transcription-polymerase chain reaction. Western blot analysis was used to confirm protein expression of PD-L1. We also tested the functional role of PD-L1 and PD-1 in antigen presentation. Furthermore, we stained mouse kidney transplants with rejection for the expression of the PD-1 ligands. RESULTS: We found that PD-L1 but not PD-L2 was weakly expressed on unstimulated TEC. Upon stimulation with IFN-gamma, a dose dependent upregulation of PD-L1 expression was observed. Blockade of the PD-L1/PD 1 pathway with monoclonal antibodies in antigen presentation assays uncovered an inhibitory role of this ligand system in Th1 and Th2 cell activation. Staining for PD-L1 was strong in proximal and distal tubules in mouse kidney transplants with rejection, whereas staining of normal kidneys and syngenic mouse kidney transplants did not reveal PD-L1 expression. PD-L2 was not observed in normal or rejected mouse kidneys. CONCLUSIONS: These data demonstrate that PD-L1 is an inducible renal tubular epithelial antigen that negatively regulates T-cell responses elicited by IFN-gamma-stimulated TEC. We speculate that the PD-1/PD-L1 pathway may play a role in protecting the epithelium from immune-mediated tubulointerstitial injury. PMID- 15353580 TI - Higher offspring survival among Tibetan women with high oxygen saturation genotypes residing at 4,000 m. AB - Here we test the hypothesis that high-altitude native resident Tibetan women with genotypes for high oxygen saturation of hemoglobin, and thus less physiological hypoxic stress, have higher Darwinian fitness than women with low oxygen saturation genotypes. Oxygen saturation and genealogical data were collected from residents of 905 households in 14 villages at altitudes of 3,800-4,200 m in the Tibet Autonomous Region along with fertility histories from 1,749 women. Segregation analysis confirmed a major gene locus with an autosomal dominant mode of inheritance for high oxygen saturation levels, associated with a 10% higher mean. Oxygen saturation genotypic probability estimators were then used to calculate the effect of the inferred oxygen saturation locus on measures of fertility, in a subsample of 691 women (20-59 years of age and still married to their first husbands, those with the highest exposure to the risk of pregnancy). The genotypic probability estimators were not significantly associated with the number of pregnancies or live births. The high oxygen saturation genotypic mean offspring mortality was significantly lower, at 0.48 deaths compared with 2.53 for the low oxygen saturation homozygote, because of lower infant mortality. Tibetan women with a high likelihood of possessing one to two alleles for high oxygen saturation had more surviving children. These findings suggest that high altitude hypoxia is acting as an agent of natural selection on the locus for oxygen saturation of hemoglobin by the mechanism of higher infant survival of Tibetan women with high oxygen saturation genotypes. PMID- 15353581 TI - Phenotypic conversion of human mammary carcinoma cells by autocrine human growth hormone. AB - We report here that autocrine production of human growth hormone (hGH) results in a phenotypic conversion of mammary carcinoma cells such that they exhibit the morphological and molecular characteristics of a mesenchymal cell, including expression of fibronectin and vimentin. Autocrine production of hGH resulted in reduced plakoglobin expression and relocalization of E-cadherin to the cytoplasm, leading to dissolution of cell-cell contacts and decreased cell height. These phenotypic changes were accompanied by an increase in cell motility, elevated activity of specific matrix metalloproteinases, and an acquired ability to invade a reconstituted basement membrane. Forced expression of plakoglobin significantly decreased mammary carcinoma cell migration and invasion stimulated by autocrine hGH. In vivo, autocrine hGH stimulated local invasion of mammary carcinoma cells concomitant with a prominent stromal reaction in comparison with well delineated and capsulated growth of mammary carcinoma cells lacking autocrine production of hGH. Thus, autocrine production of hGH by mammary carcinoma cells is sufficient for generation of an invasive phenotype. Therapeutic targeting of autocrine hGH may provide a mechanistic approach to prevent metastatic extension of human mammary carcinoma. PMID- 15353582 TI - Biography of Zdenek P. Bazant. PMID- 15353583 TI - Regulation of chromosome stability by the histone H2A variant Htz1, the Swr1 chromatin remodeling complex, and the histone acetyltransferase NuA4. AB - NuA4, the only essential histone acetyltransferase complex in Saccharomyces cerevisiae, acetylates the N-terminal tails of histones H4 and H2A. Affinity purification of NuA4 revealed the presence of three previously undescribed subunits, Vid21/Eaf1/Ydr359c, Swc4/Eaf2/Ygr002c, and Eaf7/Ynl136w. Experimental analyses revealed at least two functionally distinct sets of polypeptides in NuA4: (i) Vid21 and Yng2, and (ii) Eaf5 and Eaf7. Vid21 and Yng2 are required for bulk histone H4 acetylation and are functionally linked to the histone H2A variant Htz1 and the Swr1 ATPase complex (SWR-C) that assembles Htz1 into chromatin, whereas Eaf5 and Eaf7 have a different, as yet undefined, role. Mutations in Htz1, the SWR-C, and NuA4 cause defects in chromosome segregation that are consistent with genetic interactions we have observed between the genes encoding these proteins and genes encoding kinetochore components. Because SWR-C dependent recruitment of Htz1 occurs in both transcribed and centromeric regions, a NuA4/SWR-C/Htz1 pathway may regulate both transcription and centromere function in S. cerevisiae. PMID- 15353584 TI - The roles of latex and the vascular bundle in morphine biosynthesis in the opium poppy, Papaver somniferum. AB - The opium poppy, Papaver somniferum, is one of mankind's oldest medicinal plants. Opium poppy today is the commercial source of the narcotic analgesics morphine and codeine. Along with these two morphinans, opium poppy produces approximately eighty alkaloids belonging to various tetrahydrobenzylisoquinoline-derived classes. It has been known for over a century that morphinan alkaloids accumulate in the latex of opium poppy. With identification of many of the enzymes of alkaloid biosynthesis in this plant, biochemical data suggested involvement of multiple cell types in alkaloid biosynthesis in poppy. Herein the immunolocalization of five enzymes of alkaloid formation in opium poppy is reported: (R,S)-3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase central to the biosynthesis of tetrahydroisoquinoline-derived alkaloids, the berberine bridge enzyme of the sanguinarine pathway, (R,S)-reticuline 7-O-methyltransferase specific to laudanosine formation, and salutaridinol 7-O-acetyltransferase and codeinone reductase, which lead to morphine. In capsule and stem, both O methyltransferases and the O-acetyltransferase are found predominantly in parenchyma cells within the vascular bundle, and codeinone reductase is localized to laticifers, the site of morphinan alkaloid accumulation. In developing root tip, both O-methyltransferases and the O-acetyltransferase are found in the pericycle of the stele, and the berberine bridge enzyme is localized to parenchyma cells of the root cortex. Laticifers are not found in developing root tip, and, likewise, codeinone reductase was not detected. These results provide cell-specific localization that gives a coherent picture of the spatial distribution of alkaloid biosynthesis in opium poppy. PMID- 15353585 TI - Hematopoietic contribution to skeletal muscle regeneration by myelomonocytic precursors. AB - Adult bone marrow-derived cells can participate in muscle regeneration after bone marrow transplantation. In recent studies a single hematopoietic stem cell (HSC) was shown to give rise to cells that not only reconstituted all of the lineages of the blood, but also contributed to mature muscle fibers. However, the relevant HSC derivative with this potential has not yet been definitively identified. Here we use fluorescence-activated cell sorter-based protocols to test distinct hematopoietic fractions and show that only fractions containing c-kit(+) immature myelomonocytic precursors are capable of contributing to muscle fibers after i.m. injection. Although these cells belong to the myeloid lineage, they do not include mature CD11b(+) myelomonocytic cells, such as macrophages. Of the four sources of mature macrophages tested that were derived either from monocytic culture, bone marrow, peripheral blood after granulocyte colony-stimulating factor mobilization, or injured muscle, none contributed to muscle. In addition, after transplantation of bone marrow isolated from CD11b-Cre-transgenic mice into the Cre-reporter strain (Z/EG), no GFP myofibers were detected, demonstrating that macrophages expressing CD11b do not fuse with myofibers. Irrespective of the underlying mechanisms, these data suggest that the HSC derivatives that integrate into regenerating muscle fibers exist in the pool of hematopoietic cells known as myelomonocytic progenitors. PMID- 15353586 TI - On the road to selenocysteine. PMID- 15353587 TI - Sfp1 is a stress- and nutrient-sensitive regulator of ribosomal protein gene expression. AB - Yeast cells modulate their protein synthesis capacity in response to physiological needs through the transcriptional control of ribosomal protein (RP) genes. Here we demonstrate that the transcription factor Sfp1, previously shown to play a role in the control of cell size, regulates RP gene expression in response to nutrients and stress. Under optimal growth conditions, Sfp1 is localized to the nucleus, bound to the promoters of RP genes, and helps promote RP gene expression. In response to inhibition of target of rapamycin (TOR) signaling, stress, or changes in nutrient availability, Sfp1 is released from RP gene promoters and leaves the nucleus, and RP gene transcription is down regulated. Additionally, cells lacking Sfp1 fail to appropriately modulate RP gene expression in response to environmental cues. We conclude that Sfp1 integrates information from nutrient- and stress-responsive signaling pathways to help control RP gene expression. PMID- 15353588 TI - Molecular basis for catecholaminergic neuron diversity. AB - Catecholaminergic neurons control diverse cognitive, motor, and endocrine functions and are associated with multiple psychiatric and neurodegenerative disorders. We present global gene-expression profiles that define the four major classes of dopaminergic (DA) and noradrenergic neurons in the brain. Hypothalamic DA neurons and noradrenergic neurons in the locus coeruleus display distinct group-specific signatures of transporters, channels, transcription, plasticity, axon-guidance, and survival factors. In contrast, the transcriptomes of midbrain DA neurons of the substantia nigra and the ventral tegmental area are closely related with <1% of differentially expressed genes. Transcripts implicated in neural plasticity and survival are enriched in ventral tegmental area neurons, consistent with their role in schizophrenia and addiction and their decreased vulnerability in Parkinson's disease. The molecular profiles presented provide a basis for understanding the common and population-specific properties of catecholaminergic neurons and will facilitate the development of selective drugs. PMID- 15353589 TI - CD26 up-regulates expression of CD86 on antigen-presenting cells by means of caveolin-1. AB - CD26 is a T cell costimulatory molecule with dipeptidyl peptidase IV activity in its extracellular region. We previously reported that recombinant soluble CD26 enhanced T cell proliferation induced by the recall antigen tetanus toxoid (TT). However, the mechanism involved in this enhancement is not yet elucidated. We now demonstrate that CD26 binds Caveolin-1 on antigen-presenting cells, and that residues 201-211 of CD26 along with the serine catalytic site at residue 630 contribute to binding to caveolin-1 scaffolding domain. In addition, after CD26 caveolin-1 interaction on TT-loaded monocytes, caveolin-1 is phosphorylated, which links to activate NF-kappaB, followed by up-regulation of CD86. Finally, reduced caveolin-1 expression on monocytes inhibits CD26-mediated CD86 up regulation and abrogates CD26 effect on TT-induced T cell proliferation. Taken together, these results strongly suggest that CD26-caveolin-1 interaction plays a role in the up-regulation of CD86 on TT-loaded monocytes and subsequent engagement with CD28 on T cells, leading to antigen-specific T cell activation. PMID- 15353590 TI - Counting primes, groups, and manifolds. AB - Let Lambda=SL(2)(Z) be the modular group and let c(n)(Lambda) be the number of congruence subgroups of Lambda of index at most n. We prove that lim(n--> infinity )(log c(n)(Lambda)/((log n)(2)/log log n))=(3-2(sqrt)2)/4. The proof is based on the Bombieri-Vinogradov "Riemann hypothesis on the average" and on the solution of a new type of extremal problem in combinatorial number theory. Similar surprisingly sharp estimates are obtained for the subgroup growth of lattices in higher rank semisimple Lie groups. If G is such a Lie group and Gamma is an irreducible lattice of G it turns out that the subgroup growth of Gamma is independent of the lattice and depends only on the Lie type of the direct factors of G. It can be calculated easily from the root system. The most general case of this result relies on the Generalized Riemann Hypothesis, but many special cases are unconditional. The proofs use techniques from number theory, algebraic groups, finite group theory, and combinatorics. PMID- 15353591 TI - Reverse transcriptase activity innate to DNA polymerase I and DNA topoisomerase I proteins of Streptomyces telomere complex. AB - Replication of Streptomyces linear chromosomes and plasmids proceeds bidirectionally from a central origin, leaving recessed 5' termini that are extended by a telomere binding complex. This complex contains both a telomere protecting terminal protein (Tpg) and a telomere-associated protein that interacts with Tpg and the DNA ends of linear Streptomyces replicons. By using histidine-tagged telomere-associated protein (Tap) as a scaffold, we identified DNA polymerase (PolA) and topoisomerase I (TopA) proteins as other components of the Streptomyces telomere complex. Biochemical characterization of these proteins indicated that both PolA and TopA exhibit highly efficient reverse transcriptase (RT) activity in addition to their predicted functions. Although RT activity innate to other DNA-dependent DNA polymerases has been observed previously, its occurrence in a topoisomerase is unprecedented. Deletion mapping and sequence analysis showed that the RT activity of Streptomcyces TopA resides in a peptide region containing motifs that are absent from most bacterial topoisomerases but are highly conserved in a novel subfamily of eubacterial topoisomerases found largely in Actinobacteria. Within one of these motifs, and essential to the RT function of Streptomyces TopA, is an Asp-Asp doublet sequence required also for the RT activities of human immunodeficiency virus and eukaryotic cell telomerases. PMID- 15353592 TI - Different functional roles of T1R subunits in the heteromeric taste receptors. AB - The T1R receptors, a family of taste-specific class C G protein-coupled receptors, mediate mammalian sweet and umami tastes. The structure-function relationships of T1R receptors remain largely unknown. In this study, we demonstrate the different functional roles of T1R extracellular and transmembrane domains in ligand recognition and G protein coupling. Similar to other family C G protein-coupled receptors, the N-terminal Venus flytrap domain of T1R2 is required for recognizing sweeteners, such as aspartame and neotame. The G protein coupling requires the transmembrane domain of T1R2. Surprisingly, the C-terminal transmembrane domain of T1R3 is required for recognizing sweetener cyclamate and sweet taste inhibitor lactisole. Because T1R3 is the common subunit in the sweet taste receptor and the umami taste receptor, we tested the interaction of lactisole and cyclamate with the umami taste receptor. Lactisole inhibits the activity of the human T1R1/T1R3 receptor, and, as predicted, blocked the umami taste of l-glutamate in human taste tests. Cyclamate does not activate the T1R1/T1R3 receptor by itself, but potentiates the receptor's response to l glutamate. Taken together, these findings demonstrate the different functional roles of T1R3 and T1R2 and the presence of multiple ligand binding sites on the sweet taste receptor. PMID- 15353593 TI - Inactivation of a bacterial virulence pheromone by phagocyte-derived oxidants: new role for the NADPH oxidase in host defense. AB - Quorum sensing triggers virulence factor expression in medically important bacterial pathogens in response to a density-dependent increase in one or more autoinducing pheromones. Here, we show that phagocyte-derived oxidants target these autoinducers for inactivation as an innate defense mechanism of the host. In a skin infection model, expression of phagocyte NADPH oxidase, myeloperoxidase, or inducible nitric oxide synthase was critical for defense against a quorum-sensing pathogen, Staphylococcus aureus, but not for defense against a quorum sensing-deficient mutant. A virulence-inducing peptide of S. aureus was inactivated in vitro and in vivo by reactive oxygen and nitrogen intermediates, including HOCl and ONOO(-). Inactivation of the autoinducer prevented both the up-regulation of virulence gene expression and the downstream sequelae. MS analysis of the inactivated peptide demonstrated that oxidation of the C-terminal methionine was primarily responsible for loss of activity. Treatment of WT but not NADPH oxidase-deficient mice with N-acetyl methionine to scavenge the inhibitory oxidants increased in vivo quorum sensing independently of the bacterial burden at the site of infection. Thus, oxidant-mediated inactivation of an autoinducing peptide from S. aureus is a critical innate defense mechanism against infection with this pathogen. PMID- 15353594 TI - Potential for neural regeneration after neurotoxic injury in the adult mammalian retina. AB - It has long been believed that the retina of mature mammals is incapable of regeneration. In this study, using the N-methyl-D-aspartate neurotoxicity model of adult rat retina, we observed that some Muller glial cells were stimulated to proliferate in response to a toxic injury and produce bipolar cells and rod photoreceptors. Although these newly produced neurons were limited in number, retinoic acid treatment promoted the number of regenerated bipolar cells. Moreover, misexpression of basic helix-loop-helix and homeobox genes promoted the induction of amacrine, horizontal, and rod photoreceptor specific phenotypes. These findings demonstrated that retinal neurons regenerated even in adult mammalian retina after toxic injury. Furthermore, we could partially control the fate of the regenerated neurons with extrinsic factors or intrinsic genes. The Muller glial cells constitute a potential source for the regeneration of adult mammalian retina and can be a target for drug delivery and gene therapy in retinal degenerative diseases. PMID- 15353595 TI - Synergistic interactions of lipids and myelin basic protein. AB - This report describes force measurements and atomic force microscope imaging of lipid-protein interactions that determine the structure of a model membrane system that closely mimics the myelin sheath. Our results suggest that noncovalent, mainly electrostatic and hydrophobic, interactions are responsible for the multilamellar structure and stability of myelin. We find that myelin basic protein acts as a lipid coupler between two apposed bilayers and as a lipid "hole-filler," effectively preventing defect holes from developing. From our protein-mediated-adhesion and force-distance measurements, we develop a simple quantitative model that gives a reasonably accurate picture of the molecular mechanism and adhesion of bilayer-bridging proteins by means of noncovalent interactions. The results and model indicate that optimum myelin adhesion and stability depend on the difference between, rather than the product of, the opposite charges on the lipid bilayers and myelin basic protein, as well as on the repulsive forces associated with membrane fluidity, and that small changes in any of these parameters away from the synergistically optimum values can lead to large changes in the adhesion or even its total elimination. Our results also show that the often-asked question of which membrane species, the lipids or the proteins, are the "important ones" may be misplaced. Both components work synergistically to provide the adhesion and overall structure. A better appreciation of the mechanism of this synergy may allow for a better understanding of stacked and especially myelin membrane structures and may lead to better treatments for demyelinating diseases such as multiple sclerosis. PMID- 15353596 TI - A nonpolio enterovirus with respiratory tropism causes poliomyelitis in intercellular adhesion molecule 1 transgenic mice. AB - Coxsackievirus A21 (CAV21) is classified within the species Human enterovirus C (HEV-C) of the Enterovirus genus of picornaviruses. HEV-C share striking homology with the polioviruses (PV), their closest kin among the enteroviruses. Despite a high level of sequence identity, CAV21 and PV cause distinct clinical disease typically attributed to their differential use of host receptors. PV cause poliomyelitis, whereas CAV21 shares a receptor and a propensity to cause upper respiratory tract infections with the major group rhinoviruses. As a model for CAV21 infection, we have developed transgenic mice that express human intercellular adhesion molecule 1, the cell-surface receptor for CAV21. Surprisingly, CAV21 administered to these mice via the intramuscular route causes a paralytic condition consistent with poliomyelitis. The virus appears to invade the CNS by retrograde axonal transport, as has been demonstrated to occur in analogous PV infections. We detected human intercellular adhesion molecule 1 expression on both transgenic mouse and human spinal cord anterior horn motor neurons, indicating that members of HEV-C may share PV's potential to elicit poliomyelitis in humans. PMID- 15353597 TI - The magnetic polarity stratigraphy of the Mauch Chunk Formation, Pennsylvania. AB - Three sections of Chesterian Mauch Chunk Formation in Pennsylvania have been studied paleomagnetically to determine a Late Mississippian magnetic polarity stratigraphy. The upper section at Lavelle includes a conglomerate with abundant red siltstone rip-up clasts that yielded a positive conglomerate test. All samples were subjected to progressive thermal demagnetization to temperatures as high as 700 degrees C. Two components of magnetization were isolated: a synfolding "B" component and the prefolding "C" component. The conglomerate test is positive, indicating that the C component was acquired very early in the history of the sediment. A coherent pattern of magnetic polarity reversals was identified. Five magnetozones were identified in the upper Lavelle section, which yields a pattern that is an excellent match with the pattern of reversals obtained from the upper Mauch Chunk at the original type section of the Mississippian/Pennsylvanian boundary at Pottsville, PA. The frequency of reversals in the upper Mississippian, as identified in the Mauch Chunk Formation, is approximately one to two per million years, which is an average for field reversal through time. PMID- 15353598 TI - NF-kappa B-mediated repression of growth arrest- and DNA-damage-inducible proteins 45alpha and gamma is essential for cancer cell survival. AB - The NF-kappaB/IkappaB signaling pathway is a critical regulator of cell survival in cancer. Here, we report that combined down-regulation of growth arrest- and DNA-damage-inducible proteins (GADD)45alpha and gamma expression by NF-kappaB is an essential step for various cancer types to escape programmed cell death. We demonstrate that inhibition of NF-kappaB in cancer cells results in GADD45alpha- and gamma-dependent induction of apoptosis and inhibition of tumor growth. Inhibition of GADD45alpha and gamma in cancer cells by small interfering RNA abrogates apoptosis induction by the inhibitor of NF-kappaB and blocks c-Jun N terminal kinase activation, whereas overexpression of GADD45alpha and gamma activates c-Jun N-terminal kinase and induces apoptosis. These results establish an unambiguous role for the GADD45 family as an essential mediator of cell survival in cancer cells with implications for cancer chemotherapy and novel drug discovery. PMID- 15353599 TI - Ultrafast hydration dynamics in protein unfolding: human serum albumin. AB - We report studies of unfolding and ultrafast hydration dynamics of the protein human serum albumin. Unique in this study is our ability to examine different domains of the same protein and the intermediate on the way to the unfolded state. With femtosecond resolution and site-selective labeling, we isolate the dynamics of domains I and II of the native protein, domain I of the intermediate at 2 M guanidine hydrochloride, and the unfolded state at 6 M of the denaturant. For studies of unfolding, we used the fluorophores, acrylodan (covalently bound to Cys-34 in domain I) and the intrinsic tryptophan (domain II), whereas for hydration dynamics, we probed acrylodan and prodan; the latter is bound to domain II. From the time-dependent spectra and the correlation functions, we obtained the time scale of dynamically ordered water: 57 ps for the more stable domain I and 32 ps for the less stable domain II, in contrast to approximately 0.8 ps for labile, bulk-type water. This trend suggests an increased hydrophilic residues water interaction of domain I, contrary to some packing models. In the intermediate state, which is characterized by essentially intact domain I and unfolded domain II, the dynamics of ordered water around domain I is nearly the same (61 ps) as that of native state (57 ps), whereas that in the unfolded protein is much shorter (13 ps). We discuss the role of this fluidity in the correlation between stability and function of the protein. PMID- 15353600 TI - The inositol polyphosphate 5-phosphatase Ocrl associates with endosomes that are partially coated with clathrin. AB - The subcellular localization of Ocrl, the inositol polyphosphate 5-phosphatase that is mutated in Lowe syndrome, was investigated by fluorescence microscopy. Ocrl was localized to endosomes and Golgi membranes along with clathrin, giantin, the mannose 6-phosphate receptor, transferrin, and the early endosomal antigen 1 endosomal marker in fixed cells. The endosomal localization of Ocrl was confirmed by live-cell time-lapse microscopy in which we monitored the dynamics of Ocrl on endosomes. GST binding assays show that Ocrl interacts with the clathrin terminal domain and the clathrin adaptor protein AP-2. Our findings suggest a role for Ocrl in endosomal receptor trafficking and sorting. PMID- 15353601 TI - Dynamic alterations of membrane tethers stabilize leukocyte rolling on P selectin. AB - Leukocytes rolling on selectins extrude thin membrane tethers that might stabilize rolling velocities despite marked alterations in wall shear stress. To test this hypothesis, we used differential interference contrast videomicroscopy to visualize formation and breakage of membrane tethers as neutrophils rolled on P-selectin under flow. Neutrophils rapidly increased tether number as wall shear stress rose and decreased tether number as wall shear stress declined. Membrane tethers invariably accompanied slower, more uniform rolling steps that translated into lower mean rolling velocities and variances in velocity. Unexpectedly, neutrophils, but not fixed cells or microspheres bearing selectin ligands, rolled progressively more slowly and uniformly over time. Scanning electron microscopy revealed that neutrophils developed larger, more complex tether structures as they rolled for longer periods. These data provide evidence that neutrophils stabilize selectin-mediated rolling by rapidly adjusting tether number in response to changes in wall shear stress. Gradual remodeling of tether architecture may further reduce rolling velocities, facilitating integrin dependent deceleration and arrest on inflamed vascular surfaces. PMID- 15353602 TI - The crystal structure of ribosomal chaperone trigger factor from Vibrio cholerae. AB - Trigger factor is a molecular chaperone that is present in all species of eubacteria. It binds to the ribosomal 50S subunit near the translation exit tunnel and is thought to be the first protein to interact with nascent polypeptides emerging from the ribosome. The chaperone has a peptidyl-prolyl cis trans isomerase (PPIase) activity that catalyzes the rate-limiting proline isomerization in the protein-folding process. We have determined the crystal structure of nearly full-length trigger factor from Vibrio cholerae by x-ray crystallography at 2.5-A resolution. The structure is composed of two trigger factor molecules related by a noncrystallographic two-fold symmetry axis. The monomer has an elongated shape and is folded into three domains: an N-terminal domain I that binds to the ribosome, a central domain II that contains PPIase activity, and a C-terminal domain III. The active site of the PPIase domain is occupied by a loop from domain III, suggesting that the PPIase activity of the protein could be regulated. The dimer interface is formed between domains I and III and contains residues of mixed properties. Further implications about dimerization, ribosome binding, and other functions of trigger factor are discussed. PMID- 15353603 TI - A Populus EST resource for plant functional genomics. AB - Trees present a life form of paramount importance for terrestrial ecosystems and human societies because of their ecological structure and physiological function and provision of energy and industrial materials. The genus Populus is the internationally accepted model for molecular tree biology. We have analyzed 102,019 Populus ESTs that clustered into 11,885 clusters and 12,759 singletons. We also provide >4,000 assembled full clone sequences to serve as a basis for the upcoming annotation of the Populus genome sequence. A public web-based EST database (POPULUSDB) provides digital expression profiles for 18 tissues that comprise the majority of differentiated organs. The coding content of Populus and Arabidopsis genomes shows very high similarity, indicating that differences between these annual and perennial angiosperm life forms result primarily from differences in gene regulation. The high similarity between Populus and Arabidopsis will allow studies of Populus to directly benefit from the detailed functional genomic information generated for Arabidopsis, enabling detailed insights into tree development and adaptation. These data will also valuable for functional genomic efforts in Arabidopsis. PMID- 15353604 TI - Blue light stimulated luminescence in calcium fluoride, its characteristics and implications in radiation dosimetry. AB - Strong optically stimulated luminescence (OSL), stimulated by blue light, has been observed, for the first time, in natural calcium fluoride (CaF(2)) phosphor, used as thermoluminescence dosemeters. Traps responsible for all three thermoluminescence (TL) peaks appearing upto 300 degrees C, i.e. 126, 196 and 264 degrees C, were observed to contribute to the blue light stimulated luminescence (BLSL) in CaF(2). When the areas under the respective curves (BLSL and TL) were measured, the BLSL measured at room temperature was found to be approximately 1.8 times the TL output of the dosimetry peak that appeared at approximately 264 degrees C. However, when measured after thermally annealing the first two TL peaks, the BLSL signal was found to be 0.26 times the TL output of the dosimetry peak at 264 degrees C. This paper describes the characteristics of the BLSL signal using linearly modulated OSL after subjecting the phosphor to different annealing treatments. Feasibility studies to assess the usefulness of BLSL in CaF(2) for environmental radiation monitoring are also discussed. PMID- 15353605 TI - Decision support systems in nuclear emergencies: a scenario-based comparison of domestic and reference tools. AB - The article reports selective results of a comparison between RODOS-an emerging decision support system for the management of nuclear emergencies in Europe developed by an international research consortium under EEC aegis, and a resident software package developed and maintained for similar purposes at IFIN-HH, Bucharest. Reproducible similarity patterns obtained in the output data distributions provide for simple normalising procedures that may ensure convergent radiological assessments. When properly consolidated on a sufficient scenario casuistry, such procedures could lend a certain resilience to domestic decision support tools over the interim lead time required by the full implementation of RODOS, or other major league, internationally accepted reference systems. PMID- 15353606 TI - Hourly indoor radon measurements in a research house. AB - This paper reports and discusses the behaviour of radon concentration with time in an uninhabited dwelling. The relationship between variations in radon concentrations and indoor-outdoor temperatures and wind intensity has also been discussed. Radon concentration was measured hourly in a house located at a height of 800 m in the Lombard Prealps, at the top of the Valassina valley. The wind velocity and indoor-outdoor temperatures were measured by means of a meteorological station located on the terrace of the house. The data were analysed using the LBL model for indoor-outdoor air exchange and the models for the indoor accumulation of radon due to exhalation from building materials and pressure-driven infiltrations located underground. The role of wind and indoor outdoor temperatures were analysed. The agreement of measurements with modelling clearly demonstrates the importance of the different sources of indoor radon. As the investigation was conducted in an uninhabited house, the measurements were not affected by the behaviour of people, e.g. opening and closing of windows. Measurements of the outdoor atmospheric concentrations of (222)Rn provide an index of the atmospheric stability, the formation of thermal inversions and convective turbulence. PMID- 15353607 TI - A new approach to studying angiogenesis in rheumatoid arthritis by means of power Doppler ultrasonography and measurement of serum vascular endothelial growth factor. AB - OBJECTIVE: To evaluate angiogenesis as an essential component of pannus formation and cartilage destruction in rheumatoid arthritis (RA) using power Doppler ultrasonography (PDUS) and serum vascular endothelial growth factor (VEGF) measurement. METHODS: Twenty-one RA patients with a painful and swollen wrist and 12 healthy controls were examined with ultrasound. By means of standard scans, vascularity near and inside the joint capsule was visualized with PDUS. Two trained investigators performed sonography. Representative video clips were stored and read by two independent investigators, under blinded conditions, with regard to the microvascular Doppler flow being either inside or outside the joint capsule and with respect to a qualitative estimate of the intensity of blood flow, according to a grading from 1 to 3. Serum levels of VEGF were measured with a standard quantitative sandwich ELISA. RESULTS: The power Doppler mode identified increased synovial microvascular blood flow inside the joint capsule in 17 of 21 RA patients (81%) vs one of the healthy controls. We found large variation in serum VEGF levels in RA patients and in healthy controls. The degree of synovial vascularity determined by PDUS showed no correlation with the immediate serum VEGF level in the same patient. CONCLUSION: The high correlation between intra-articular microvascular power Doppler flow and clinical synovitis in RA patients (P<0.0001) indicates that PDUS may be helpful in studying the role of synovial blood vessels in rheumatoid inflammation. PMID- 15353608 TI - A longitudinal study of disease activity and functional status in a hospital cohort of patients with ankylosing spondylitis. AB - OBJECTIVE: To evaluate changes in functional status and disease activity and their determinants in patients with ankylosing spondylitis (AS) attending hospital, using the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and the Bath Ankylosing Spondylitis Functional Index (BASFI). METHODS: Patients completed BASDAI and BASFI questionnaires annually from 1996 to 2001. Demographic and clinical data were collected. The mean first and last recorded scores were compared. The change per year and area under the curve per year for the BASDAI and BASFI were calculated. Relationships between demographic, clinical and longitudinal BASDAI/BASFI data were examined. Subgroup analyses were performed using the cross-sectional and longitudinal data. RESULTS: Two hundred and seventy nine BASDAI and 322 BASFI questionnaires were analysed. The BASFI increased [mean change 6.15, 95% confidence interval (CI) 1.9, 10.3, P = 0.005] but the BASDAI did not (mean change 0.87, 95% CI -3.96, 5.7, P = 0.71). First recorded scores were the best predictors of the cumulative scores per year. Patients with peripheral joint (P = 0.01) and hip (P<0.001) disease had higher mean BASFI scores. Males (P<0.001) and patients with spinal disease alone (P = 0.0014), iritis (P = 0.005) and late-onset AS (P = 0.002) became more functionally impaired over time. CONCLUSIONS: Disease activity in this AS cohort remained relatively constant but there was functional decline. Initial BASDAI/FI can predict a severe disease course. PJD patients with peripheral joint disease were more functionally impaired, but deteriorated less than spinal disease alone patients. Iritis and late onset disease may be severity markers for functional impairment. PMID- 15353609 TI - Putative anti-muscarinic antibodies cannot be detected in patients with primary Sjogren's syndrome using conventional immunological approaches. AB - OBJECTIVE: To determine whether autoantibodies directed against muscarinic M3 receptors are present in the serum of patients with primary Sjogren's syndrome (pSS), and if so whether these autoantibodies inhibit secretion by intact salivary acinar cells. METHODS: IgG was purified by affinity chromatography using protein G from sera collected from 15 patients with pSS. Antibody detection was by Western blotting, whole-cell enzyme-linked immunosorbent assay (ELISA) and immunoblotting. The antisecretory activity of the IgG faction was determined using fura-2 microfluorimetry to measure changes in intracellular Ca(2+) activity ([Ca(2+)](i)) in human and mouse salivary gland acinar cells and in Chinese hamster ovary (CHO) cells transfected with human M3 receptors (CHO-M3). RESULTS: We found no specific M3 receptor recognition by the IgG fraction obtained from pSS patients using either Western blotting or ELISA or immunoblot techniques in which epitope conformation were preserved. Chronic exposure to pSS IgG had no effect on agonist-evoked Ca(2+) signals measured in human or mouse submandibular acinar cells or in CHO-M3 cells. However, acute application of IgG from Sjogren's syndrome patients produced a rapidly reversible reduction in the agonist stimulated elevation in [Ca(2+)](i). CONCLUSION: These data represent the first demonstration of salivary acinar cell inhibition by pSS IgG; however, this inhibition was found to be reversible. Our data also show that pSS IgG binding to M3R cannot be visualized by conventional immunological approaches. PMID- 15353610 TI - Gene expression relevant to osteoclastogenesis in the synovium and bone marrow of mature rats with collagen-induced arthritis. AB - OBJECTIVES: To investigate gene expression relevant to osteoclastogenesis in the synovium and bone marrow during the development of collagen-induced arthritis (CIA) in mature rats. METHODS: Total messenger RNAs (mRNAs) were obtained from CIA synovium and bone marrow after immunization. First, reverse transcriptase polymerase chain reactions (RT-PCR) were carried out to detect the mRNA encoding receptor activator of NF-kappaB (RANK), RANK ligand (RANKL), osteoprotegerin (OPG), tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6 and the osteoclast markers tartrate-resistant acid phosphatase (TRAP) and cathepsin K. Secondly, the genes detected clearly by RT-PCR were quantified using real-time PCR. RESULTS: In the synovium, expression of all genes was confirmed by specific single bands in RT-PCR. In real-time PCR, the expression levels of TNF-alpha, IL 1beta, IL-6, RANKL, TRAP and cathepsin K mRNA increased, whereas the expression levels of RANK and OPG were unchanged and decreased respectively. RANKL expression was highly correlated with the two osteoclast markers. In the bone marrow, RT-PCR did not clearly detect the expression of IL-6, RANKL or OPG mRNA. Quantitative real-time PCR showed that TNF-alpha, RANK and TRAP mRNA expression did not change significantly with time, and that IL-1beta and cathepsin K changed slightly compared with those in the synovium. CONCLUSIONS: In the early stages of arthritis, synovial RANKL is closely involved in osteoclastogenesis, and various changes in synovial cytokines, including down-regulation of OPG, probably accelerate osteoclast formation. In contrast, cytokine mRNA in the bone marrow showed little fluctuation. We suggest that synovial cytokines affect osteoclastogenesis not only in the synovium but in the bone marrow. PMID- 15353611 TI - Health-related quality of life in Italian patients with systemic lupus erythematosus. I. Relationship between physical and mental dimension and impact of age. AB - OBJECTIVE: To examine health-related quality of life (HRQOL) in Italian patients with systemic lupus erythematosus (SLE) and compare it with that of healthy people, and to investigate relationships among different dimensions and subscales of a generic health status measure. METHODS: The Medical Outcomes Study Short Form-36 (SF-36) was applied in a cohort of 126 consecutive SLE patients and 96 healthy controls. At the time of HRQOL testing, all patients underwent clinical and laboratory evaluation. RESULTS: Both physical (PCS) and mental (MCS) component summary scores of the SF-36 were reduced in patients compared with controls. In SLE great variability in all the subscales was observed. Significant correlations between PCS and MCS and between many different subscales were observed in patients but not in controls. The PCS was higher than MCS more frequently in controls than in SLE patients (81 vs 48.4%, P<0.00001). In SLE, HRQOL tended to worsen with age. CONCLUSION: Both PCS and MCS contribute to the decrease in HRQOL in SLE patients. In SLE the mutual interaction between these two dimensions seems to be more relevant than in healthy people. PMID- 15353612 TI - The non-thiol angiotensin-converting enzyme inhibitor quinapril suppresses inflammatory arthritis. AB - OBJECTIVES: In addition to its vasoactive effects, angiotensin II has proinflammatory properties. Angiotensin-converting enzyme (ACE) inhibitors reduce the production of angiotensin II and could therefore act as anti-inflammatory agents. Here we investigated the capacity of the ACE inhibitor quinapril to modulate inflammatory arthritis. METHODS: We studied the effect of quinapril on disease activity in mice with collagen-induced arthritis (CIA). Mice received oral quinapril (10 mg/kg/day) at the time of arthritis induction (prophylaxis protocol) or at the onset of mild arthritis (therapy protocol). Concentrations of immunoglobulin G (IgG) subtypes specific for bovine Type II collagen and TNF alpha were measured by enzyme-linked immunoassay. RESULTS: Quinapril significantly diminished the activity of CIA when given as prophylaxis or therapy (prophylaxis protocol, P<0.001; therapy protocol P=0.002). Antigen-specific IgG2a antibodies were reduced by 52% (P=0.02) in the quinapril prophylaxis protocol. Suppression of arthritis by quinapril was associated with reduced articular expression of TNF-alpha by 68% (P=0.01) in the prophylaxis protocol and 27% (P=0.06) in the therapy protocol. Quinapril therapy also inhibited expression of splenocyte TNF-alpha production following lipopolysaccharide (LPS) in vitro stimulation by 59% (P=0.02). In parallel human in vitro experiments, ACE inhibition suppressed LPS-stimulated production of TNF-alpha by monocytes. In order to confirm that the action of quinapril occurred predominantly through suppression of angiotensin II, parallel experiments with the angiotensin receptor antagonist candesartan cilexetil demonstrated that this agent also inhibited disease activity in CIA. CONCLUSIONS: These data suggest that angiotensin II is a mediator of chronic inflammation and that ACE inhibition may have therapeutic effects in human inflammatory arthritis. PMID- 15353614 TI - Advances in nuclear particle dosimetry for radiation protection and medicine. Proceedings of the Ninth Symposium on Neutron Dosimetry. September 28 - October 3, 2003. Delft, The Netherlands. PMID- 15353613 TI - Evidence-based recommendations for the role of exercise in the management of osteoarthritis of the hip or knee--the MOVE consensus. AB - OBJECTIVES: Exercise is an effective and commonly prescribed intervention for lower limb osteoarthritis (OA). Many unanswered questions remain, however, concerning the practical delivery of exercise therapy. We have produced evidence based recommendations to guide health-care practitioners. METHODS: A multidisciplinary guideline development group was formed from representatives of professional bodies to which OA is of relevance and other interested parties. Each participant contributed up to 10 propositions describing key clinical points regarding exercise therapy for OA of the hip or knee. Ten final recommendations were agreed by the Delphi technique. The research evidence for each was determined. A literature search was undertaken in the Medline, PubMed, EMBASE, PEDro, CINAHL and Cochrane databases. The methodological quality of each retrieved publication was assessed. Outcome data were abstracted and effect sizes calculated. The evidence for each recommendation was assessed and expert consensus highlighted by the allocation of two categories: (1) strength of evidence and (2) strength of recommendation. RESULTS: The first round of the Delphi process produced 123 propositions. This was reduced to 10 after four rounds. These related to aerobic and strengthening exercise, group versus home exercise, adherence, contraindications and predictors of response. The literature search identified 910 articles; 57 intervention trials relating to knee OA, 9 to hip OA and 73 to adherence. The evidence to support each proposition is presented. CONCLUSION: These are the first recommendations for exercise in hip and knee OA to clearly differentiate research evidence and expert opinion. Gaps in the literature are identified and issues requiring further study highlighted. PMID- 15353615 TI - Neutron detection and neutron dosimetry. AB - A review is presented on the research and development of inorganic scintillators for position-sensitive thermal neutron detectors to be used at new spallation neutron sources. Furthermore some recent developments in the field of fast neutron dosimetry will be discussed. PMID- 15353616 TI - Ion chamber gas-to-wall conversion factors for fast neutron dosimetry. AB - Modern ionising photon dosimetry is essentially entirely based upon gas-filled cavity determinations. For photons, ion chamber response is largely independent of photon energy almost perfectly transforming absorbed dose in the gas to the surrounding media. Absolute uncertainties are <1-2%. For fast neutron dosimetry, this is certainly not the case. Interpretation of the response of the cavity filling material, usually a gas, to the charged particle spectrum induced in the walls and interacting with the cavity gas is fraught with uncertainties. Despite these challenges, gas filled cavities surrounded by various mixtures, compounds and elements, have proved to be essential for integral determinations of the indirectly ionising neutrons, generating dosimetric quantities, such as kerma and absorbed dose. The transformation from gas response to wall dose is material dependent and varies with neutron energy. This study discusses recent advances in cavity response interpretation using the results from complex nuclear modelling of microscopic cross sections as well as estimates of secondary particle production enabling much improved cavity gas-to-wall media conversion factors. PMID- 15353617 TI - Total nucleon-nucleus reaction cross sections calculated in the frame of the cascade model. AB - Total nucleon reaction cross sections with nuclei are necessary as input data for various applications, such as transport calculations or prediction of different radiation effects. Nevertheless, experimental data until now are rather scarce and contradictory whereas reliable theoretical models are absent. The optical model of nuclear reactions, most often used for the calculation of reaction cross sections, cannot be applied for extrapolation into the regions where no experimental data exist, due to the large number of free parameters in the phenomenological optical potential. This paper presents the calculations of nucleon reaction cross sections in the frame of the new generalised cascade model. The model includes the self-consistent description of the nucleon density and potential in nuclei, accurate consideration of the refraction process by the mean field, non-locality of the nuclear interaction and reduction of the nucleon nucleon interaction cross sections in dense nuclear matter. The results of the calculations show overall good agreement with available experimental data in wide ranges of nuclei and nucleon energies from 10 MeV to 10 GeV without free model parameters. The present model may be a good base for the creation of global systematics of total nucleon reaction cross sections. PMID- 15353618 TI - Neutron calibration facilities. AB - Reliable measurement of neutron radiation is a difficult task due to the large energy range of neutrons, their complex and energy-dependent interaction mechanisms with matter and, consequently, the imperfect response characteristics of most instruments. Therefore, Calibration procedures and calibration facilities play an important role. Different types of calibration fields have been developed and made available at several institutions. The primary reference quantity used for the calibration of neutron measuring devices--area monitors, personal dosemeters, spectrometers, etc.--is the neutron fluence. This quantity is determined by appropriate experimental methods whereas dosimetric quantities are derived by applying recommended fluence-to-dose conversion coefficients. This paper summarises the basic principles underlying neutron production, the metrology employed to characterise the radiation fields and the calibration procedures employed. Examples of calibration facilities will be given, which enable routine calibrations, investigations of energy dependence and application specific calibrations. PMID- 15353619 TI - Developing a thermal neutron irradiation system for the calibration of personal dosemeters in terms of Hp(10). AB - At the ENEA Radiation Protection Institute in Bologna a thermal neutron irradiation facility is available for the calibration of neutron dosemeters. It consists of a 1 m x 1 m x 1 m polyethylene cube containing three 241Am-Be sources of about 185 GBq. The cube contains three co-axial cylindrical calibration cavities of different dimension. Due to their limited dimensions, the cavities do not allow the calibration of thermal neutron personal dosemeters in terms of Personal Dose Equivalent Hp(d), that should be carried out on the 30 cm x 30 cm x 15 cm ISO phantom. The study herewith presented was addressed at adapting the facility for external irradiation of personal dosemeters on the ISO phantom. Extensive Monte Carlo studies were carried out to characterise the neutron fluence spatial distribution along the front face of the phantom. A satisfying neutron field homogeneity within the measurement area has been obtained by means of a pyramidal polyethylene fluence flattening filter and the selection of the proper cube to phantom distance. This new irradiation set-up was experimentally tested through measurements with activation foils, according to the spatial mapping array taken from the calculations. PMID- 15353620 TI - AMANDE: a new facility for monoenergetic neutron fields production between 2 keV and 20 MeV. AB - The variation of the response of the instruments with the neutron energy has to be determined in well-characterized monoenergetic neutron fields. The AMANDE facility will deliver such neutron fields between 2 keV and 20 MeV in an experimental hall designed with metallic walls for neutron scattering minimisation. The neutrons will be produced by nuclear interaction of accelerated protons or deuterons on thin targets of selected materials. The measuring devices to be characterised will be accurately placed with a fully automated detector transport system. The energy of the neutron field will be validated by time-of flight experiments and a large set of standard detectors and fluence monitors will be used to determine the neutron fluence references. The scattered neutron fluence and dose equivalent were calculated by the MCNP Monte Carlo code at several measuring points in order to determine their contribution to the neutron field. PMID- 15353621 TI - Double-differential heavy-ion production cross sections. AB - Current computational tools used for space or accelerator shielding studies transport energetic heavy ions either using a one-dimensional straight-ahead approximation or by dissociating the nuclei into protons and neutrons and then performing neutron and proton transport using Monte Carlo techniques. Although the heavy secondary particles generally travel close to the beam direction, a proper treatment of the light ions produced in these reactions requires that double-differential cross sections should be utilised. Unfortunately, no fundamental nuclear model capable of serving as an event generator to provide these cross sections for all ions and energies of interest exists currently. Herein, we present a model for producing double-differential heavy-ion production cross sections that uses heavy-ion fragmentation yields produced by the NUCFRG2 fragmentation code coupled with a model of energy degradation in nucleus-nucleus collisions and systematics of momentum distributions to provide energy and angular dependences of the heavy-ion production. PMID- 15353622 TI - Double-differential light-ion production cross sections. AB - Codes used for space radiation shielding studies typically transport light-ions from the incident cosmic ray spectrum using either a one-dimensional straight ahead approximation or in three dimensions, often without accounting for their breakup. Light-ions are also produced by breakup of energetic heavy ions in target materials. Herein, we present cross section models that can be used to predict double-differential light-ion production cross sections. Deuteron breakup is based on a parameterization of the total reaction cross sections. Alpha fragmentation utilizes a parameterization based on an abrasion-ablation model that has been augmented by experimental data. Neutron and proton production is modelled with a quantum mechanical abrasion-ablation-coalescence model. This same model is also used to predict cross sections for triton and 3He breakup. Finally, energies and emission angles for particles other than nucleons are specified using a model of energy degradation in nucleus-nucleus collisions and systematics of momentum distributions. PMID- 15353623 TI - W values of protons in alkane-based TE gases and dimethylether in the energy range 1-100 keV: measurements and additivity models. AB - W values of tissue-equivalent gases based on propane and butane were measured for protons in the energy range 1-100 keV and compared with those of dimethylether and water vapour. The experimental values of the tissue-equivalent gas mixtures were compared with data calculated from the W values of the mixture components using five existing additivity models and two new models derived within the scope of the present work. Apart from one model--the simplest one--all models show a good agreement with the experimental data. PMID- 15353624 TI - Characterization of gamma rays existing in the NMIJ standard neutron field. AB - Our laboratory provides national standards on fast neutron fluence. Neutron fields are always accompanied by gamma rays produced in neutron sources and surroundings. We have characterised these gamma rays in the 5.0 MeV standard neutron field. Gamma ray measurement was performed using an NE213 liquid scintillator. Pulse shape discrimination was incorporated to separate the events induced by gamma rays from those by neutrons. The measured gamma ray spectra were unfolded with the HEPRO program package to obtain the spectral fluences using the response matrix prepared with the EGS4 code. Corrections were made for the gamma rays produced by neutrons in the detector assembly using the MCNP4C code. The effective dose equivalents were estimated to be of the order of 25 microSv at the neutron fluence of 10(7) neutrons cm(-2). PMID- 15353625 TI - Characteristics of the neutron field of the facility at DIN-UPM. AB - A new source facility (241Am-Be) has been installed in a bunker-type room of large dimensions. To characterise the neutron fields in the facility, detailed calculations have been made with MCNP-4C, showing the different components of the neutron radiation reaching the reference points (direct, inscattered, backscattered). The contribution from neutrons scattered in the walls to the total ambient dose equivalent remains reasonably low (<10%) in the reference points. Additionally, spectra measurements have been performed with a Bonner spheres spectrometer with a 6LiI(Eu) scintillator (0.4 phi x 0.4 cm2), UTA4 response matrix and BUNKIUT unfolding code. The calculated and experimentally obtained spectra are compared, with small differences found in the epithermal and thermal region, attributable to the concrete composition used in the calculations. The H*(10) rate has been determined from the spectra, and then compared to the reading of an active dosemeter (LB6411), with differences found lower than 8%. PMID- 15353626 TI - Neutron spectra and angular distributions of concrete-moderated neutron calibration fields at JAERI. AB - The Facility of Radiation Standards of Japan Atomic Energy Research Institute has been equipped with concrete-moderated neutron calibration fields as simulated workplace neutron fields. The fields use an 241Am-Be neutron source placed in the narrow space surrounded by concrete bricks, walls and floor. The neutron spectra and the neutron fluence rates of the fields were measured with the Bonner multi sphere spectrometer system (BMS), spherical recoil-proton proportional counters, and a liquid scintillation counter (NE-213). The results were compared with each other. The reference values of H*(10) were determined from the results of BMS. The angular distributions of neutron fluence were calculated using MCNP-4B2 to obtain the reference values of Hp(10). The calculated results show that the scattered neutrons have a wide range of incident angles. The reference Hp(10) values considered the angular distribution were found to be 10-18% smaller than those without consideration. PMID- 15353627 TI - Construction of 144, 565 keV and 5.0 MeV monoenergetic neutron calibration fields at JAERI. AB - Monoenergetic neutron calibration fields of 144, 565 keV and 5.0 MeV have been developed at the Facility of Radiation Standards of JAERI using a 4 MV Pelletron accelerator. The 7Li(p,n)7Be and 2H(d,n)3He reactions are employed for neutron production. The neutron energy was measured by the time-of-flight method with a liquid scintillation detector and calculated with the MCNP-ANT code. A long counter is employed as a neutron monitor because of the flat response. The monitor is set up where the influence of inscattered neutrons from devices and their supporting materials at a calibration point is as small as possible. The calibration coefficients from the monitor counts to the neutron fluence at a calibration point were obtained from the reference fluence measured with the transfer instrument of the primary standard laboratory (AIST), a 24.13 cm phi Bonner sphere counter. The traceability of the fields to AIST was established through the calibration. PMID- 15353628 TI - Conceptual design of spectrum changeable neutron calibration fields in JAERI/FRS. AB - Spectrum changeable neutron calibration fields are planned to be established with an accelerator installed in Japan Atomic Energy Research Institute/Facility of Radiation Standards. The neutron fields are provided by bombarding a target surrounded by a moderator, with charged particles from the accelerator. In the fields, a wide variety of neutron spectra is provided with sufficient fluence rate for the calibration of dosemeters. In this study, necessity of the field was first discussed in view of relationship between readings of existing dosemeters and true dose equivalents where the dosemeters were used. Second, test simulation of neutron spectra was carried out with the Monte Carlo technique for some arrangements with a LiF target and quasi-cylindrical moderators with different materials. The simulated spectra were summarised in terms of fluence-average energy, fluence rate and calibration factor for the dosemeters. PMID- 15353629 TI - Quasi-monoenergetic neutron reference fields in the energy range from thermal to 200 MeV. AB - Well-characterised neutron fields are a prerequisite for the investigation of neutron detectors. Partly in collaboration with external partners, the PTB neutron metrology group makes available for other users neutron reference fields covering the full energy range from thermal to 200 MeV. The specification of the neutron fluence in these beams is traceable to primary standard cross sections. PMID- 15353630 TI - Inter-laboratory test program (PTI) in 'neutron dosimetry'. AB - The Laboratory of External Dosimetry Studies and Researches (LRDE), associated laboratory to the Bureau National de Metrologie, is strongly involved in the reference establishment and calibration of instruments in neutron radiation fields provided by 241Am-Be, 252Cf and (252Cf + D2O)/Cd sources. In 1997, on the request of COFRAC, the LRDE has managed an Inter-Laboratory Test Program in 'neutron dosimetry' to compare the different standard calibration methods. The transfer instrument used was a "Harwell Monitor N91". The results of the participant were in good agreement but the associated uncertainties need to be revised by some of laboratories. According to the Mutual Recognition Arrangement signed in 1999, laboratories for metrology will have to assure the traceability of their references. In 2003-2005, the LRDE organises, with National Physical Laboratory (UK) and Physikalisch-Techische Bundesanstalt (Germany), an international comparison in 'neutron dosimetry'. Two transfer instruments, a "2202D" rate metre and a "Harwell Monitor N91", will be sent in turn to the participant laboratories for measurements at their own neutron sources facilities. PMID- 15353631 TI - Standard fields of old neutron sources--parameters and traceability. AB - Standard neutron fields of isotope neutron sources, established in the Institute of Atomic Energy, Poland, have been examined for nearly 20 y. The neutron dose equivalent and gamma dose rates were measured, applying various techniques. The neutron emission of standard neutron sources of 241Am-Be and 252Cf has been originally determined in primary standard laboratory, then checked, making use of transfer instrument calibrated in primary laboratory. The growth of neutron emission of 239Pu-Be source, used for routine calibrations, has been demonstrated. The total uncertainty of determined parameters has been discussed. The periodically repeated checks of neutron fields of standard sources with the use of transfer instrument, calibrated in primary laboratory, should provide the traceability to primary laboratory and the fields could be officially recognised. PMID- 15353632 TI - A realistic field facility to simulate reactor spectra. AB - The design construction and characterisation of a simulated realistic neutron workplace field is described. Utilising a low-energy primary source of neutrons based on the 7Li(p,n) reaction, and a heavy water moderator, a broad spectrum is produced with energies extending from thermal to about 1 MeV. The field simulates the type of spectra encountered around UK gas-cooled reactors. PMID- 15353633 TI - Characteristics of the simulated workplace neutron fields using a 252Cf source surrounded with cylindrical moderators. AB - The authors established the simulated workplace neutron fields using a 252Cf source surrounded with cylindrical moderators at the Japan Nuclear Cycle Development Institute (JNC), Tokai Works. The moderators are annular cylinders made of polymethyl methacrylate and steel. The neutron energy spectrum at the reference calibration point was evaluated from the calculations by MCNP-4B and the measurements by the Bonner multisphere spectrometer and the hydrogen-filled proportional counters. The calculated neutron spectra were in good agreements with the measured ones. These fields can provide the realistic neutron spectra similar to those encountered around the glove-boxes of the fabrication process of MOX (PuO2-UO2 mixed oxide) fuel. PMID- 15353634 TI - Monte Carlo simulation of the IRSN CANEL/T400 realistic mixed neutron-photon radiation field. AB - The calibration of dosemeters and spectrometers in realistic neutron fields simulating those encountered at workplaces is of high necessity to provide true and reliable dosimetric information to the exposed nuclear workers. The CANEL assembly was set-up at IRSN to produce such neutron fields. It comprises a depleted uranium shell, to produce fission neutrons, then iron and water to moderate them and a polyethylene duct. The new presented CANEL facility is used with 3.3 MeV neutrons. Calculations were performed with the MCNP4C code to characterise this mixed neutron-photon expanded radiation field at the position where calibrations are usually performed. The neutron fluence energy and the direction distributions were calculated and the operational quantities were derived from these distributions. The photon fluence and corresponding ambient dose equivalent were also estimated. Comparison with experimental results showed an overall good agreement. PMID- 15353635 TI - Measurement and Monte Carlo modelling of the JRC 241Am-Li(alpha,n) source spectrum. AB - The neutron energy spectrum of the JRC 241Am-Li(alpha,n) radionuclide source has been measured at the PTB in Germany using various spectrometry systems, such as Bonner spheres, proton recoil counters and NE213 liquid scintillators. The source photon spectrum has been measured and the neutron contamination due to traces of beryllium through the 9Be(alpha,n)12C reaction has been determined. The effects associated with source encapsulation and the materials it contains have been investigated previously, using Monte Carlo simulations, the various structures seen in the spectra have been explained and the anisotropy of fluence distributions in 4pi have been calculated and compared to measurements. These extensive high resolution spectrometry measurements have been coupled to Monte Carlo calculations to provide a realistic spectrum and new spectrum-averaged fluence-to-dose equivalent conversion coefficients for the actual 241Am-Li source. PMID- 15353636 TI - Characterisation of the IRSN graphite moderated Americium-Beryllium neutron field. AB - The SIGMA facility was set up at IRSN to provide thermal neutrons for metrology and dosimetry purposes. SIGMA consists of six Am-Be radioactive sources located in a 1.5 x 1.5 x 1.5 m3 graphite moderator block. The neutron field at the calibration position, situated at 50 cm from the west surface of the assembly was characterised experimentally and by Monte Carlo calculations. The thermal neutron fluence was determined by the activation of gold foils; the neutron fluence energy distribution above 240 keV was measured with proton recoil spectrometers and the neutron fluence energy distribution from thermal energies to 20 MeV was measured with a Bonner spheres spectrometer. A Monte Carlo simulation of the SIGMA assembly was undertaken using the MCNP4C code, and the calculated neutron fluence energy distribution was compared with the measurements. As a whole, the experimental data and the MCNP calculation are in a good agreement. PMID- 15353637 TI - Neutron spectrometry for radiation protection. AB - The various instruments used for neutron spectrometry can be divided into four categories. Within each of these categories, the underlying measurement principle for all devices is essentially the same. The applications of the instruments within each group to spectrometry for radiation protection are reviewed. PMID- 15353638 TI - Measurement of neutron fluence spectra up to 150 MeV using a stacked scintillator neutron spectrometer. AB - A stacked scintillator neutron spectrometer (S3N) consisting of three slabs of liquid organic scintillator is described. A pulsed beam providing a broad spectrum of neutron energies is used to determine the detection efficiency of the spectrometer as a function of incident neutron energy and to measure the pulse height response matrix of the system. Neutron spectra can then be determined for beams with any kind of time structure by unfolding pulse height spectra measured by the S3N. Examples of fluence spectrum measurements in the energy range 20-150 MeV are presented. PMID- 15353639 TI - Neutron spectra in a tissue equivalent phantom during photon radiotherapy treatment by LINACS. AB - Bremsstrahlung photon beams produced by LINAC accelerators are currently the most used radiotherapy method for tumour treatments. When the photon energy exceeds the (gamma,n) reaction threshold energy, the patient receives an undesired dose due to photoneutron production both in the accelerator head and in the human body. In this paper, a method is presented for the assessment of the photoneutron spectra produced by Giant Dipole Resonance (GDR) during cancer radiotherapy with energetic photon beams. Experimental and numerical results have been obtained for the facility at Onkologik Klinik, Lund (Sweden), which is based on an ELEKTA 18 MV LINAC. Neutron spectra are evaluated both at the patient plane and inside an anthropomorphic phantom. PMID- 15353640 TI - The response of a bonner sphere spectrometer to charged hadrons. AB - Bonner sphere spectrometers (BSSs) are employed in neutron spectrometry and dosimetry since many years. Recent developments have seen the addition to a conventional BSS of one or more detectors (moderator plus thermal neutron counter) specifically designed to improve the overall response of the spectrometer to neutrons above 10 MeV. These additional detectors employ a shell of material with a high mass number (such as lead) within the polyethylene moderator, in order to slow down high-energy neutrons via (n,xn) reactions. A BSS can be used to measure neutron spectra both outside accelerator shielding and from an unshielded target. Measurements were recently performed at CERN of the neutron yield and spectral fluence at various angles from unshielded, semi-thick copper, silver and lead targets, bombarded by a mixed proton/pion beam with 40 GeV per c momentum. These experiments have provided evidence that under certain circumstances, the use of lead-enriched moderators may present a problem: these detectors were found to have a significant response to the charged hadron component accompanying the neutrons emitted from the target. Conventional polyethylene moderators show a similar behaviour but less pronounced. These secondary hadrons interact with the moderator and generate neutrons, which are in turn detected by the counter. To investigate this effect and determine a correction factor to be applied to the unfolding procedure, a series of Monte Carlo simulations were performed with the FLUKA code. These simulations aimed at determining the response of the BSS to charged hadrons under the specific experimental situation. Following these results, a complete response matrix of the extended BSS to charged pions and protons was calculated with FLUKA. An experimental verification was carried out with a 120 GeV per c hadron beam at the CERF facility at CERN. PMID- 15353641 TI - Development of active environmental and personal neutron dosemeters. AB - For neutron dosimetry in the radiation environment surrounding nuclear facilities, two types of environmental neutron dosemeters, the high-sensitivity rem counter and the high-sensitivity multi-moderator, the so-called Bonner ball, have been developed and the former is commercially available from Fuji Electric Co. By using these detectors, the cosmic ray neutrons at sea level have been sequentially measured for about 3 y to investigate the time variation of neutron spectrum and ambient dose equivalent influenced by cosmic and terrestrial effects. Our Bonner ball has also been selected as the neutron detector in the International Space Station and has already been used to measure neutrons in the US experimental module. The real time wide-range personal neutron dosemeter which uses two silicon semiconductor detectors has been developed for personal dosimetry and is commercially available from Fuji Electric Co. This dosemeter has good characteristics, fitted to the fluence-to-dose conversion factor in the energy range from thermal energies to several tens of mega-electron-volts and is now widely used in various nuclear facilities. PMID- 15353642 TI - Advanced passive detectors for neutron dosimetry and spectrometry. AB - Different neutron detectors have been developed in the past which exploit electrical and electrochemical processes in plastic foils and thin-film capacitors (namely metal-oxide-silicon devices) to trigger avalanche processes, which greatly facilitate the detection of neutron-induced charged particles. These detectors are: (i) spark-replica counter of neutron-induced fission fragment holes in plastic films, thin-film breakdown counter of neutron-induced fission fragments, and electrochemically etched detectors of neutron-induced recoils in plastic foils. The major shortcomings of damage-track detectors for the measurement of low neutron fluencies, such as those of cosmic ray neutrons at civil aviation altitudes, are their large and unpredictable background and their small signal-to-noise ratio. These shortcomings have been overcome respectively by using long exposure times and large detector areas and counting coincidence track events on matched pairs of detectors even for a few-micron-long tracks such as those of neutron recoils. The responses of all these detectors have been analysed both with neutrons with energy up to approximately 200 MeV and protons up to tens of gigaelectron volts. Applications of these detectors for the cosmic ray neutron dosimetry and/or spectrometry will be mentioned. PMID- 15353643 TI - Angle dependence of response characteristics of neutron survey instruments. AB - Neutron area survey instruments are designed to have an approximately isotropic response. In practice, the response cannot be perfectly isotropic for instruments that do not have spherical symmetry, and for all instruments it is modified by the inclusion of batteries, electronics, handles, etc. This affects the ability of the survey instrument to measure accurately an isotropic dose equivalent quantity. Measurements of the angle dependence of response for four of the most commonly used designs of survey instrument (Harwell 0949, Mark 7 NRM, NM2 and Studsvik 2202D) have been performed in a low-scatter room using radionuclide and monoenergetic neutron sources. The Monte Carlo code MCNP has been used to model the responses and to investigate their sensitivity to the polyethylene density, counting gas pressure and other manufacturing tolerances. Preliminary modelling results are presented here. PMID- 15353644 TI - Passive detectors for neutron personal dosimetry: state of the art. AB - Passive, solid-state detectors still dominate the field of neutron personal dosimetry, mainly thanks to their low cost, high reliability and elevated throughput. However, the recent appearance in the market of several electronic personal dosemeters for neutrons presents a challenge to the exclusive use of passive systems for primary or official dosimetry. This scenario drives research and development activities on passive dosemeters towards systems offering greater accuracy of response and lower detection limits. In addition, further applications and properties of the passive detectors, which are not met by the electronic devices, are also being explored. In particular, extensive investigations are in progress on the use of solid-state detectors for aviation and space dosimetry, where high-energy neutron fields are encountered. The present situation is also stimulating an acceleration in the development of international standards on performance and test requirements for passive dosimetry systems, which can expedite significantly the implementation of techniques in commercial personal dosimetry services. Upcoming standards will cover thermoluminescence albedo dosemeters, etched-track detectors, superheated emulsions and direct ion storage chambers, attesting to the level of maturity reached by these techniques. This work reviews the developments in the field of passive neutron dosimetry emerged since the previous Neutron Dosimetry Symposium, reporting on the current status of the subject and indicating the direction of ongoing research. PMID- 15353645 TI - Numerical and experimental results of the operational neutron dosemeter 'Saphydose-N'. AB - Since 1993, the Institute for Radiological Protection and Nuclear Safety (IRSN) has lead, in association with Electricite de France (EDF), a R&D study of a neutron personal electronic dosemeter. This dosemeter, called 'Saphydose-N', is manufactured by the SAPHYMO company. This paper presents first the optimisation of some detector components using Monte Carlo calculations, and second the test of the manufactured Saphydose-N under radiation following the IEC 1323 standard's recommendations for active personal neutron dosemeters. The measurements with the manufactured dosemeter were performed on the one hand at PTB (Physikalisch Technische Bundesanstalt) in mono-energetic neutron fields and, on the other hand at IRSN in neutron fields generated by a thermal facility (SIGMA), radionuclide ISO sources and a realistic spectrum (CANEL/T400). The manufactured dosemeter Saphydose-N was also tested during measurement campaigns of the European programme EVIDOS ('Evaluation of Individual Dosimetry in Mixed Neutron and Photon Radiation Fields') at different nuclear workplaces. The study showed that Saphydose-N complies with the recommendations of standard IEC 1323 and can be used at any workplace with no previous knowledge of the neutron field characteristics. PMID- 15353646 TI - Improved neutron detection by gamma-ray spectroscopy. AB - Most high-efficiency interactions for neutron detection are with thermal neutrons, and the detection of the prompt gamma radiation emitted as a result of thermal neutron absorption in specific materials was sometimes used. These materials are named converters, the most widespread being 10B, 113Cd or 73Ge (the last two materials are detector materials themselves for CdZnTe and HPGe detectors). However, for these materials, the main gamma-ray energies are relatively low (in the region 473-600 keV), and there are interferences from other gamma rays present in background. Different combinations of moderator converter-detector systems were employed to optimise the system efficiency and signal-to-background ratio, using B, Cd, Gd and Cl converters and HPGe, NaI(Tl) and BGO detectors. When using a Cl converter, the 1164.7 keV peak seems to be a good choice when a HPGe spectrometer is used. A very practical material containing chlorine is PVC. PMID- 15353647 TI - Present status of the personal neutron dosemeter based on direct ion storage. AB - In this paper the present status of the Direct Ion Storage Neutron (DIS-N) prototype dosemeter (RADOS) is described. The separation of neutron from photon dose equivalent has been improved by adding tin shieldings. The neutron energy response has been changed by additional plastic covers containing 40% B4C in order to reduce the over-response to thermal neutrons. The responses of the dosemeters were determined for standard photon and neutron fields (monoenergetic neutrons, neutron sources and simulated workplace fields). Irradiations in real workplaces were also performed. The dependence of the neutron response on the angle of incidence was measured for different neutron sources. PMID- 15353648 TI - Development of a portable high-energy neutron spectrometer. AB - The design of a portable high-energy (20-800 MeV) neutron spectrometer based on CsI or BaF2 is described. The particle discrimination properties of these scintillators allow the light-ion spallation products (p, d, t and alpha) from neutron interactions to be identified uniquely. One or more of the resulting pulse-height spectra can be unfolded to reveal the incident neutron spectrum. Dosimetric quantities can then be calculated based on the unfolded spectrum. Due to the high stopping power of these scintillators, modest-sized crystals are suitable for this application. Combined with advances in electronics, a lightweight instrument capable of on-line particle discrimination with a real time display of neutron-induced count rate is feasible. Preliminary experimental data are presented, and the importance of validating MCNPX-generated response functions is discussed. A brief discussion on future work follows. PMID- 15353649 TI - Development of a fast neutron spectrometer composed of silicon-SSD and position sensitive proportional counters. AB - A new fast neutron spectrometer has been developed. The spectrometer is composed of a silicon surface barrier detector and three position-sensitive proportional counters with methane gas working as counting gas and a radiator. A collimated incident neutron interacts with a hydrogen atom in the methane gas to generate a recoil proton. The position information on the path of the recoil proton obtained from the three position-sensitive proportional counters gives the recoil angle. In the meanwhile, the energy of the recoil protons is measured with the three proportional counters and the silicon surface barrier detector. The characteristics of the spectrometer were evaluated with a monoenergetic neutron beam. The best energy resolution was 1.8% for 5.0 MeV neutrons. PMID- 15353650 TI - Hardness ratios of different neutron spectra. AB - Extensive data have been gathered in the past on the response of different detectors, based on the registration of neutron-induced fissions in bismuth, gold, tantalum and thorium by the spark-replica counter and the thin film breakdown counter. These detectors make it possible to exploit the excellent characteristics of the fission reactions for the measurements of high-energy neutrons. Most of the investigations have been carried out at the quasi monoenergetic neutron beam facility at The Svedberg Laboratory-TSL of the Uppsala University in cooperation with the Khlopin Radium Institute (KRI). The responses of different fission detectors in the neutron energy range 35-180 MeV have been evaluated: a region where the predictive power of available nuclear reaction models and codes is not reliable yet. For neutron energy >200 MeV, the fission detector responses have been derived from the data of the proton fission cross sections. By using the ratio of the responses of these detectors, a simple and accurate way to evaluate the spectrum hardness can be obtained, thus providing a tool to obtain spectral information needed for neutron dosimetry without the need to know the entire spectrum. Extensive data have been already obtained for the high-energy neutron spectrum from the CERN concrete facility. In the present paper, the measured values of the response ratios for different fissile detectors exposed at the CERN facility are compared with those calculated for the spectra from the same facility and from different altitudes in the atmosphere, respectively. PMID- 15353651 TI - Development and application of CVD diamond detectors to 14 MeV neutron flux monitoring. AB - CVD diamond is an interesting material for radiation detection, its atomic number (Z = 6) is close to that of soft tissues (Z = 7.1) and it can also work in harsh environments. Since many years CVD diamond films have been grown at the Faculty of Engineering, Rome 'Tor Vergata' University, and in 1998 a collaboration with ENEA Fusion Division was established to develop fast neutron monitors to be used in fusion tokamak environment. In this paper the first test of a 120 microm thick polycrystalline CVD diamond detector used for monitoring 14.7 MeV neutrons emission produced with the Frascati Neutron Generator (FNG) is reported. The detector operates in air and in pulse mode. The time irradiation profiles recorded with the CVD diamond detector were compared with those recorded by the standard monitors available at FNG (SSD, fission chamber, NE-213). Good stability and capability to operate in neutron flux up to 1.5 x 10(8) n cm(-2) s(-1) was observed. The radiation hardness property was also investigated using a 460 microm thick film and these results are also reported. PMID- 15353652 TI - Measurement of energy and direction distribution of neutron and photon fluences in workplace fields. AB - Within the EU Project EVIDOS, a spectrometer with 24 silicon detectors mounted on the surface of a polyethylene sphere is used for the determination of the energy and direction distribution of neutrons and photons. It has been characterized with respect to neutron radiation with energies from thermal up to 15 MeV and to photon radiation with energies from 65 keV to 6 MeV. The first measurements described here were performed in the simulated workplace field, CANEL, at Cadarache, with the purpose of checking the instrument and the unfolding procedures. PMID- 15353653 TI - A new area multidetector dosemeter for mixed n-gamma fields. AB - A new instrument to assess neutron ambient doses has been designed and constructed. In its design, spectrometric capabilities have been implemented that allow to take into account the energy spectrum of the neutron field in the evaluation of the operational magnitude, ambient dose equivalent, H*(10). This dosemeter is based on the moderation-absorption technique and can be employed over a wide range of energies from thermal to 20 MeV. It consists of a spherical shaped polyethylene moderator with a set of thermoluminescence dosemeters (TLDs) inserted in different positions of its interior to evaluate the external neutron energy spectrum. At this moment the system uses pairs 6LiF:Mg,Ti (TLD-600) and 7LiF:Mg,Ti (TLD-700) thermoluminescence dosemeters for a better gamma discrimination. The dosemeter response matrix was calculated using the MCNP4C Monte Carlo code (MC). The viability of the dosemeter for area dosemetry has been examined experimentally showing its capabilities over a wide range of energies. PMID- 15353654 TI - ANDI-03: a genetic algorithm tool for the analysis of activation detector data to unfold high-energy neutron spectra. AB - The thresholds of (n,xn) reactions in various activation detectors are commonly used to unfold the neutron spectra covering a broad energy span, i.e. from thermal to several hundreds of MeV. The saturation activities of the daughter nuclides (i.e. reaction products) serve as the input data of specific spectra unfolding codes, such as SAND-II and LOUHI-83. However, most spectra unfolding codes, including the above, require an a priori (guess) spectrum to starting up the unfolding procedure of an unknown spectrum. The accuracy and exactness of the resulting spectrum primarily depends on the subjectively chosen guess spectrum. On the other hand, the Genetic Algorithm (GA)-based spectra unfolding technique ANDI-03 (Activation-detector Neutron DIfferentiation) presented in this report does not require a specific starting parameter. The GA is a robust problem solving tool, which emulates the Darwinian Theory of Evolution prevailing in the realm of biological world and is ideally suited to optimise complex objective functions globally in a large multidimensional solution space. The activation data of the 27Al(n,alpha)24Na, 116In(n,gamma)116mIn, 12C(n,2n)11C and 209Bi(n,xn)(210-x)Bi reactions recorded at the high-energy neutron field of the ISIS Spallation source (Rutherford Appleton Laboratory, UK) was obtained from literature and by applying the ANDI-03 GA tool, these data were used to unfold the neutron spectra. The total neutron fluence derived from the neutron spectrum unfolded using GA technique (ANDI-03) agreed within +/-6.9% (at shield top level) and +/-27.2% (behind a 60 cm thick concrete shield) with the same unfolded with the SAND-II code. PMID- 15353655 TI - Development of neutron-monitor detector using liquid organic scintillator coupled with 6Li + ZnS(Ag) Sheet. AB - A phoswitch-type detector has been developed for monitoring neutron doses in high energy accelerator facilities. The detector is composed of a liquid organic scintillator (BC501A) coupled with ZnS(Ag) sheets doped with 6Li. The dose from neutrons with energies above 1 MeV is evaluated from the light output spectrum of the BC501A by applying the G-function, which relates the spectrum to the neutron dose directly. The dose from lower energy neutrons, on the other hand, is estimated from the number of scintillations emitted from the ZnS(Ag) sheets. Characteristics of the phoswitch-type detector were studied experimentally in some neutron fields. It was found from the experiments that the detector has an excellent property of pulse-shape discrimination between the scintillations of BC501A and the ZnS(Ag) sheets. The experimental results also indicate that the detector is capable of reproducing doses from thermal neutrons as well as neutrons with energies from one to several tens of megaelectronvolts (MeV). PMID- 15353656 TI - Development and testing of an active area neutron dosemeter. AB - This paper describes the design, development and testing of an active area neutron dosemeter (AAND). The classic moderator and central detector is retained but in AAND this arrangement is augmented by small thermal neutron detectors positioned within the moderating body. The outputs from these detectors are combined using an appropriately weighted linear superposition to fit both the ambient dose equivalent and the radiation weighting factor. Experimental verifications of both the modelled detector energy reponses and the overall AAND response are given. In the relatively soft D2O moderated 252Cf spectra, the AAND determined both the H*(10) and mean radiation weighting factor to better than +10%. PMID- 15353657 TI - A new approach to the dosimetry of mixed radiation using a recombination chamber. AB - A new method of handling data derived from saturation curves of a recombination chamber is proposed. The method involves formation and extrapolation of a graph y(x), where y and x are simple, well-defined functions of the ratio of the ionisation current of the recombination chamber irradiated in the radiation field investigated to that in the field of a reference gamma radiation, using the same set of voltages, applied consecutively to the chamber. It makes it possible to determine separately the low linear energy transfer (LET) and high-LET components of an absorbed dose and also other important dosimetric quantities characterising a mixed radiation field. PMID- 15353658 TI - A measuring system with a recombination chamber for neutron dosimetry around medical accelerators. AB - A measuring system for dosimetry of neutrons generated around medical electron accelerators is proposed. The system consists of an in-phantom tissue-equivalent recombination chamber and associated electronics for automated control and data acquisition. A second ionization chamber serves as a monitor of photon radiation. Two quantities are determined by the recombination chamber--the total absorbed dose and the recombination index of radiation quality. The ambient dose equivalent, H*(10), or neutron absorbed dose in an appropriate phantom, can be then derived from the measured values. Tests of the system showed that a 0.5% dose contribution of neutrons to the absorbed dose of photons could be detected and estimated under laboratory conditions. Preliminary tests at the 15 MV Varian Clinac 2300C/D medical accelerator confirmed that the measuring system could be used under clinical conditions. The H*(10) of the mixed radiation was determined with an accuracy of approximately 10%. PMID- 15353659 TI - Development of a pMOSFET sensor with a Gd converter for low energy neutron dosimetry. AB - A pMOSFET having a 10 microm thick Gadolinium (Gd) layer has been invented as a slow neutron sensor. When slow neutrons are incident to the Gd layer, conversion electrons, which generate electron-hole pairs in the SiO2 layer of the pMOSFET, are generated by a neutron capture process. The holes are easily trapped in the oxide and act as positive-charge centres in the oxide. Due to the induced charges, the threshold turn-on voltage of the pMOSFET is changed. The developed sensors were tested at a neutron beam port of the HANARO research reactor and a 60Co irradiation facility to investigate slow neutron response and gamma ray contamination, respectively. The resultant voltage change was proportional to the accumulated neutron dose and it was very sensitive to slow neutrons. Moreover, ionising radiation contamination was negligible. It can also be used in a mixed radiation field by subtracting the voltage change of a pMOSFET without Gd from that of the Gd-pMOSFET. PMID- 15353660 TI - Novel developments in the MOSFET dosemeter for neutron dosimetry applications. AB - The feasibility of large-geometry Metal Oxide Semiconductor Field Effect Transistor (MOSFET) devices has been assessed for both active and passive neutron dosimetry and use in radiotherapy environments. Neutron sensitivity has been enhanced with the use of polymeric cement surrounding the gate region. Neutron activation via nuclear interaction processes is a potential problem with conventionally packaged and fabricated devices. To overcome this problem, a unique low-activation device design is described. Standard Dual in-Line devices, modified with polymeric cement and boron loaded cement have been exposed to gamma rays (60Co) and neutrons (gamma-ray shielded 252Cf) to provide neutron sensitivity estimates. The results show that the neutron sensitivity can be increased by a factor of approximately three by the use of a thin layer of polymeric cement over the gate region. Essentially zero activation is observed in the activation-reduced design compared with 1000 cps in the conventional design MOSFET when both are exposed under identical conditions to a neutron field from a gamma-ray shielded 252Cf isotopic source. PMID- 15353661 TI - The electronic neutron/photon dosemeter PTB DOS-2002. AB - During the last few years, PTB has developed the electronic dosemeter DOS-2002. It is of an especially simple design (1 silicon detector) and detects the photon and neutron personal dose equivalent with a low detection threshold of 0.016 and 10 microSv, respectively. Its dosimetric characteristics have been determined in neutron fields with energies ranging from thermal energies up to 15 MeV and in photon fields with mean energies from 65 keV to 7 MeV. It can be used in a wide temperature region from -20 degrees C to +50 degrees C, shows almost no interference in electromagetic fields but is still shock sensitive. PMID- 15353662 TI - Performance of a cylindrical tissue-equivalent proportional counter for use in neutron monitoring. AB - Tissue-equivalent proportional counters (TEPC) allow the measurements of the absorbed dose and the ambient dose equivalent for neutron fields. A device based on this approach, called NAUSICAA((1,2)), has already been developed by IRSN to be used in high energy neutron fields for space applications. The response of this detector underestimates significantly the dose equivalent at low energies (several hundred keV) which represent the major component of neutron fields at workplaces in the nuclear industry. A counter with a similar geometry (cylindrical detector) and a lower gas pressure was studied in order to simulate a 1 microm biological site. In 2003, the performance of the device was further improved by adding a small amount of 3He to the tissue-equivalent gas (propane based) in order to increase the response for the lower energies of neutrons. Three amplification circuits were used to cover lineal energy range from 10(-1) to 10(4) keV microm(-1). Tests were performed in monoenergetic neutron and source fields. This paper presents the experimental results obtained with this change. PMID- 15353663 TI - Development of a TL detector for neutron measurement by CaSO4:Dy phosphors. AB - Personal neutron dosimetry is quite a difficult area because a neutron is always accompanied with gamma radiation, which is required of a capability for mixed field dosimetry. CaSO4:Dy phosphor is known to have a very high sensitivity to gamma radiation, but the neutron capture cross section of the constituents of CaSO4:Dy are so small that the interactions between the thermal neutron and the phosphor are rare. One method to improve the neutron interaction is by introducing an impurity ion with a large thermal neutron captures cross section into the phosphor to act as a neutron target centre such as 6Li. In neutron-gamma mixed radiation fields, if two detectors for the 6Li-7Li compounds embedded CaSO4:Dy thermoluminescent (TL) pellets are used, a 6Li-compound embedded pellet can detect the neutron and gamma radiations together, and the other pellet can only detect the gamma radiation. Recently, the Korea Atomic Energy Research Institute (KAERI) has developed a new type of CaSO4:Dy TL materials embedded with phosphorous (KCT-300) to detect beta and gamma radiation with a very high sensitivity. This paper presents the development of CaSO4:Dy TL pellets embedded with 6Li compound for a thermal neutron measurement, and the detection method of the neutron and gamma dose in mixed fields with CaSO4:Dy TL pellets embedded with a 6Li compound (KCT-306) and CaSO4:Dy TL pellets embedded with a 7Li compound (KCT-307) is introduced. The net neutron sensitivity of CaSO4:Dy TL pellets embedded with 6Li compound developed in this study is about two times higher than that of the TLD-600 (Harshaw Chemical) dosemeter which is available commercially. PMID- 15353664 TI - On fading corrections for LiF:Mg,Ti irradiated by thermal neutrons. AB - The phenomena of preirradiation and postirradiation fading of LiF:Mg,Ti phosphors are considered a significant practical problem in routine dosimetry because of the uncertainties introduced, especially for mixed neutron-gamma dosimetry, as different fading corrections may be necessary for the gamma and neutron contributions. We have investigated if the region of interest (ROI) method is applicable to minimizing the fading effect for thermal neutron irradiation as it is for gamma irradiation. By employing the ROI method, which consists of integration between channel 31 below the glow curve maximum and channel 16 above the glow curve maximum, a constant dose value was obtained for the period of 90 d. Thus, by employing the ROI method, a fading independent evaluation of the thermal neutron dose can be made. PMID- 15353665 TI - Introduction of a thermal response to the DSTL PADC personal neutron dosemeter. AB - The response of the Defence Science and Technology Laboratory (DSTL) PADC personal neutron dosemeter is strongly dependent upon neutron energy, with a range of 300-500 tracks per cm2 per mSv for energies between 1 and 5 MeV. Below 1 MeV the response drops off sharply. This lack of sensitivity is undesirable when the dosemeter is employed with the softened fission spectra encountered in the workplace. In order to incorporate a thermal response, a polypropylene converter doped with LiF has been placed directly in front of the PADC elements. Tritons produced in the thermal neutron reaction 6Li (n,t)alpha at 2.7 MeV will then penetrate the PADC, leaving a trail of damage. The reaction rate within the converter has been calculated using MCNP for thermal neutrons and a range of higher energies, while transport of the tritons is modelled using the SRIM/TRIM package to determine the resultant track density and depth distribution. The modelling and experimental work have demonstrated that a concentration of 0.2% natural lithium by weight results in a track density in a thermal field comparable with that produced per unit personal dose equivalent by neutrons greater than 1 MeV in the standard dosemeter. Additional MCNP modelling has demonstrated that the dosemeters' albedo response to intermediate energy neutrons can be enhanced considerably by placing a boron-doped shield in front of the converter and increasing its lithium concentration. PMID- 15353666 TI - Dosimetry of heavy charged particles with thermoluminescence detectors--models and applications. AB - Lithium fluoride thermoluminescent (TL) detectors, with Li-7 isotope and various activators (MTS-7 LiF:Mg,Ti, MTT-7 LiF:Mg,Ti with enhanced Ti concentration and MCP-7 LiF:Mg,Cu,P) were used for dosimetry of heavy charged particles, within the ICHIBAN experiment. The microdosimetric model has been applied to calculate detection efficiency, eta, relative to gamma-ray dose, of these detectors after proton and heavy charged particle (HCP) irradiation for ion charges ranging from Z = 1 to Z = 6 and in the energy range from 0.3 to 20 MeV amu(-1). The calculated ratio eta(MCP-7)/eta(MTS-7) lies in the range between 0.2 and 1.0 for protons and between 0.2 and 0.4 for HCP with Z > 1. The calculated value of eta(MTT 7)/eta(MTS-7) for protons was found range between 1.0 and 1.45 and, for Z > 1, between 1.3 and 2. These relationships can be applied to derive information about the 'effective LET' in an unknown HCP field and to correct the TLD readings for dose evaluation. PMID- 15353667 TI - Fast-neutron OSL sensitivity of thallium-doped ammonium salts. AB - The main problem in selecting suitable thermoluminescent (TL) materials for fast neutron dosimetry is finding a material that is both tissue-equivalent and not damaged upon heating. Optically stimulated luminescence (OSL) avoids the need to heat the materials and allows the use of materials with a high content of hydrogen (responsible for 90% of the absorbed dose of fast-neutrons). The choice of studying the ammonium salts for their OSL properties was based on the calculation of their neutron kerma factor. A constant ratio of an ammonium salt's kerma coefficients to the tissue's kerma coefficients (in the fast-neutron range) is a prerequisite for a similar energy response to neutrons, i.e. tissue equivalency. The salts studied are NH4Br and (NH4)2SiF6 both doped with Tl+. This paper describes the OSL properties of Tl(+)-doped NH4Br and (NH4)2SiF6 after exposure to 14.5 MeV neutrons to explore their potential for developing new, tissue-equivalent OSL materials suitable for fast-neutron dosimetry. The relative neutron sensitivity, k, defined as the ratio of the sensitivity of the material to neutrons to its sensitivity to gamma rays, has been determined for 14.5 MeV neutrons and varies between k = 0.15 and k = 0.5. The latter value is a factor 2.5 higher than that found for known TL materials (k < or = 0.2). A drawback of these materials is the fast fading of the OSL signal. PMID- 15353668 TI - Superheated emulsions as high-energy neutron dosemeters. AB - Superheated emulsions being inexpensive, easy to fabricate, and having tissue equivalent composition make them as one of the popular neutron dosemeters. One more advantage is that they can be made insensitive to gamma rays by the choice of the sensitive liquid. It is observed that the response of commercially available bubble detector to neutron decreases above 20 MeV while its response is roughly flat in the 0.1-15 MeV region. This restricts its application as a dosemeter to high-energy neutrons. The response of bubble detector from Bubble Technology Industries, has been observed by using Pb-breeder for high-energy neutrons from different facilities in Japan. It is observed that 2-3 cm Pb breeder is effective in increasing the response of the detector to the nominal value. Theoretical calculation using MCNPX code indicates an increase in neutrons in the energy range of 0.1-10 MeV with Pb-breeder. The present work indicates the possibility of using the bubble detector as a dosemeter to high-energy neutron using a Pb-breeder of proper thickness. PMID- 15353669 TI - Comparison of imaging plates with track detectors for fast-neutron dosimetry. AB - Imaging plate (IP) radiation detectors are widely used in industrial radiography, medical imagery and autoradiography. When an IP is exposed to ionising radiation, some of the energy is absorbed to form a latent image. The energy stored, which is proportional to the dose received, can be liberated by a selective optical stimulation and collected to reconstitute the distribution of the ionising radiation on the IP. In this work, IPs for use in fast-neutron measurements are characterised. The response of our IP dosemeters in conjunction with their reading system was found to be linear in dose between 75 microSv and 10 mSv. This performance is compared with those of dosemeters based on the plastic track detectors PN3 and CR-39. PMID- 15353670 TI - A TLD-based personal dosemeter system for aircrew monitoring. AB - Evaluation of LiF:Mg,Ti thermoluminescence dosemeters (TLDs) according to the high-temperature ratio (HTR) method enables the determination of the dose-average linear energy transfer (LET), the mean quality factor and the dose equivalent in mixed radiation fields of unknown composition. The neutron contribution is assessed by the Extended Pair method calibrated in the CERN-EU High-Energy Reference Field (CERF). The advantages of the small passive detectors as an easy to-handle monitoring system for in-flight surveillance are demonstrated by measurements on-board north-bound and trans-equatorial flights. The experimental results are compared with calculations by the well-established CARI code. PMID- 15353671 TI - Passive in-flight neutron spectrometry by means of bonner spheres. AB - The spectral distribution of the neutron fluence rate was determined on-board a series of eight trans-atlantic flights between Cologne (Germany) and Washington, DC (USA), using a passive Bonner Sphere spectrometer based on thermoluminescence detectors. Contrary to the commonly applied active systems, the passive instrument facilitates a complete discrimination of gamma ray and charged particle-induced events in the detector count rate. The system was calibrated in the CERN-EU High-Energy Reference Field (CERF). The measured spectra are compared with FLUKA Monte Carlo simulations and show excellent agreement. PMID- 15353672 TI - A mobile TEPC-based system to measure the contributions to H*(10) at flight altitudes. AB - A very promising method to measure the ambient dose equivalent H*(10) at flight altitudes is to use Tissue Equivalent Proportional Counters (TEPC). The measured quantity is the lineal energy, y, which can be converted into equivalent dose as a good estimate of H*(10). According to the lineal energy transfer (LET) spectra one may even extract information about the composition of the radiation field. A new system was developed by adding a surrounding coincidence detector (CACS: Coincidence/Anti-Coincidence Shield) that allows one to identify the primary particle, which deposits energy in the TEPC, as neutral or charged. The entire system was calibrated in the neutron and high-energy photon reference fields at PTB. One of the results of these measurements is, the use of low- and high-LET calibration factors when performing measurements in mixed radiation fields. The TEPC/CACS system is now operated on-board aircrafts as a fixed or mobile dosimetry system. PMID- 15353673 TI - Aircrew exposure monitoring: results of 2001 to 2003 studies. AB - Aircrew exposure represents one of the recent subjects of occupational individual dosimetry. Since 1991 many new results have been found; there is however a need to gather further data on this exposure and its variation with geomagnetic position, solar activity and flight route parameters. Since 2001, many individual and six long-term monitoring programmes have been conducted onboard aircraft of Czech Airlines (CSA). In these programmes, a Si-diode spectrometer was fixed in an aircraft. Together with it, passive dosemeters thermoluminescent detector, track-etch based neutron dosemeter linear energy transfer and spectrometer) were exposed. More than 700 regular commercial flights were monitored in this manner. CSA supplied us also with full navigation data, which allowed us to calculate the exposure levels using EPCARD 3.2 and CARI6 codes. Direct experimental readings obtained with the detectors mentioned above were interpreted on the basis of calibrations in on-Earth reference fields and compared with calculated data. A satisfactory correlation between all sets of data was observed. PMID- 15353674 TI - Study of the ratio of non-neutron to neutron dose components of cosmic radiation at typical commercial flight altitudes. AB - CIEMAT, in close co-operation with Iberia Airlines, carried out an extensive programme of in-flight measurements, covering both hemispheres, during the years 2001 and 2002. Although the instrumentation onboard included different active devices, the results presented here were obtained from a polyethylene/tungsten moderated rem meter (SWENDI2; Eberline) and an ionisation chamber (RSS-131; Reuter-Stokes) used for measuring the ambient dose equivalent due to the neutron and the non-neutron components of cosmic radiation, respectively. This paper presents a study of each of the dose components mentioned as a function of the vertical cut-off rigidity and the flight altitude. The ratio between the two components is also presented to determine the variations in cosmic radiation composition as a function of the aforementioned parameters. The experimental results have also been compared with those predicted by the code EPCARD3.2 for the non-neutron and the neutron components of the ambient dose equivalent. PMID- 15353675 TI - In-flight measured and predicted ambient dose equivalent and latitude differences on effective dose estimates. AB - The results from 2 years (2001-2002) of experimental measurements of in-board radiation doses received at IBERIA commercial flights are presented. The routes studied cover the most significant destinations and provide a good estimate of the route doses as required by the new Spanish regulations on air crew radiation protection. Details on the experimental procedures and calibration methods are given. The experimental measurements from the different instruments (Tissue Equivalent Proportional Counter and the combination of a high pressure ion chamber and a high-energy neutron compensated rem-counter) and their comparison with the predictions from some route-dose codes (CARI-6, EPCARD 3.2) are discussed. In contrast with the already published data, which are mainly focused on North latitudes over parallel 50, many of the data presented in this work have been obtained for routes from Spain to Central and South America. PMID- 15353676 TI - Averaged particle dose conversion coefficients in air crew dosimetry. AB - The MCNPX Monte Carlo code was used to calculate energy-dependent fluence-to effective dose conversion coefficients for neutrons, protons, electrons, photons, charged pions and muons. The FLUKA Monte Carlo code was used to calculate the spectral particle fluences of secondary cosmic rays for different altitudes, and for different combinations of solar modulation and vertical cut-off rigidity parameters. The energy-averaged fluence-to-dose conversion coefficients were obtained by folding the particle fluence spectra with the conversion coefficients for effective dose and ambient dose equivalent. They show a slight dependence on altitude, solar activity and location in the geomagnetic field. PMID- 15353677 TI - Investigations of doses on board commercial passenger aircraft using CR-39 and thermoluminescent detectors. AB - Measurements of cosmic radiation dose rates (from the neutron and the non-neutron components) on board passenger aircraft were performed using environmental packages with thermoluminescent TL and CR-39 etched track detectors. The packages were calibrated at the CERN-EU high-energy Reference Field Facility and evaluated at the Institute of Nuclear Physics in Krakow (TL + CR-39) and at the German Aerospace Centre in Cologne (CR-39). Detector packages were exposed on board passenger aircraft operated by LOT Polish Airlines, flown between February and May 2001. The values of effective dose rate determined, averaged over the measuring period, ranged between 2.9 and 4.4 microSv h(-1). The results of environmental measurements agreed to within 10% with values calculated from the CARI-6 code. PMID- 15353678 TI - TEPC measurements in commercial aircraft. AB - The collaborative project involving the Mullard Space Science Laboratory (MSSL), Virgin Atlantic Airways (VAA), the UK Civil Aviation Authority (CAA) and the UK National Physical Laboratory (NPL) has been performing tissue-equivalent proportional counter measurements of cosmic ray doses in commercial aircraft since January 2000. In that time data have been recorded on over 700 flights, including over 150 flights with Air New Zealand (ANZ). This substantial set of data from the southern hemisphere is an ideal complement to the London-based measurements performed primarily on VAA flights. Although some ANZ data remains to be analysed, dose information from 111 flights has been compared with the CARI and EPCARD computer codes. Overall, the agreement between the measurements and EPCARD was excellent (within 1% for the total ambient dose equivalent), and the difference in the total effective doses predicted by EPCARD and CARI was <5%. PMID- 15353679 TI - The energy spectrum of cosmic-ray induced neutrons measured on an airplane over a wide range of altitude and latitude. AB - Crews of high-altitude aircraft are exposed to radiation from galactic cosmic rays (GCRs). To help determine such exposures, the Atmospheric Ionizing Radiation Project, an international collaboration of 15 laboratories, made simultaneous radiation measurements with 14 instruments on a NASA ER-2 high-altitude airplane. The primary instrument was a sensitive extended-energy multisphere neutron spectrometer. Its detector responses were calculated for energies up to 100 GeV using the radiation transport code MCNPX 2.5.d with improved nuclear models and including the effects of the airplane structure. New calculations of GCR-induced particle spectra in the atmosphere were used to correct for spectrometer counts produced by protons, pions and light nuclear ions. Neutron spectra were unfolded from the corrected measured count rates using the deconvolution code MAXED 3.1. The results for the measured cosmic-ray neutron spectrum (thermal to >10 GeV), total neutron fluence rate, and neutron dose equivalent and effective dose rates, and their dependence on altitude and geomagnetic cut-off agree well with results from recent calculations of GCR-induced neutron spectra. PMID- 15353680 TI - Evaluation of solid state nuclear track detector stacks exposed on the international space station. AB - The aim of the study was to investigate the contribution of secondary neutrons to the total dose inside the International Space Station (ISS). For this purpose solid-state nuclear track detector (SSNTD) stacks were used. Each stack consisted of three CR-39 sheets. The first and second sheets were separated by a Ti plate, and the second and third sheets sandwiched a Lexan polycarbonate foil. The neutron and proton responses of each sheet were studied through MC calculations and experimentally, utilising monoenergetic protons. Seven stacks were exposed in 2001 for 249 days at different locations of the Russian segment 'Zvezda'. The total storage time before and after the exposure onboard was estimated to be seven months. Another eight stacks were exposed at the CERF high-energy neutron field for calibration purposes. The CR-39 detectors were evaluated in four steps: after 2, 6, 12 and 20 h etching in 6 N NaOH at 70 degrees C (VB = 1.34 microm h( 1)). All the individual tracks were investigated and recorded using an image analyser. The stacks provided the averaged neutron ambient dose equivalent (H*) between 200 keV and 20 MeV, and the values varied from 39 to 73 microSv d(-1), depending on the location. The Lexan detectors were used to detect the dose originating from high-charge and high-energy (HZE) particles. These results will be published elsewhere. PMID- 15353681 TI - Single event upsets in semiconductor devices induced by highly ionising particles. AB - A new model of single event upsets (SEUs), created in memory cells by heavy ions and high energy hadrons, has been developed. The model takes into account the spatial distribution of charge collection efficiency over the cell area not considered in previous approaches. Three-dimensional calculations made by the HADRON code have shown good agreement with experimental data for the energy dependence of proton SEU cross sections, sensitive depths and other SEU observables. The model is promising for prediction of SEU rates for memory chips exposed in space and in high-energy experiments as well as for the development of a high-energy neutron dosemeter based on the SEU effect. PMID- 15353682 TI - The use of passive personal neutron dosemeters to determine the neutron dose equivalent component of radiation fields in spacecraft. AB - For the altitude range and inclination of the International Space Station (ISS), secondary neutrons can be a major contributor to dose equivalent inside a spacecraft. The exact proportion is very dependent on the amount of shielding of the primary galactic cosmic radiation and trapped particles, but is likely to lie in the range of 10-50%. Personal neutron dosemeters of simple design, processed using simple techniques developed for personal dosimetry, may be used to estimate this neutron component. PMID- 15353683 TI - Exposure of aircraft crew to cosmic radiation: on-board intercomparison of various dosemeters. AB - Owing to their professional activity, flight crews may receive a dose of some millisieverts within a year; airline passengers may also be concerned. The effective dose is to be estimated using various experimental and calculation tools. The European project DOSMAX (Dosimetry of Aircrew Exposure during Solar Maximum) was initiated in 2000 extending to 2004 to complete studies over the current solar cycle during the solar maximum phase. To compare various dosemeters in real conditions simultaneously in the same radiation field, an intercomparison was organised aboard a Paris-Tokyo round-trip flight. Both passive and active detectors were used. Good agreement was observed for instruments determining the different components of the radiation field; the mean ambient dose equivalent for the round trip was 129 +/- 10 microSv. The agreement of values obtained for the total dose obtained by measurements and by calculations is very satisfying. PMID- 15353684 TI - Cosmic radiation exposure of aircraft crew: compilation of measured and calculated data. AB - The aim of the working group has been to bring together, in particular from European research groups, the available, preferably published, experimental data and results of calculations, together with detailed descriptions of the methods of measurement and calculation. The purpose is to provide a dataset for all European Union Member States for the assessment of individual doses and/or to assess the validity of different approaches, and to provide an input to technical recommendations by the Article 31 group of experts and the European Commission. The radiation protection quantity of interest is effective dose, E (ISO), but the comparison of measurement results obtained by different methods or groups, and comparison of measurement results and the results of calculations, is done in terms of the operational quantity ambient dose equivalent, H*(10). The final report giving the results of the investigations will be published by the European Commission Directorate General Transport and Energy. This paper gives a preview of the report. PMID- 15353685 TI - New calculations of the atmospheric cosmic radiation field--results for neutron spectra. AB - The propagation of primary cosmic rays through the Earth's atmosphere and the energy spectra of the resulting secondary particles have been calculated using the Monte Carlo transport code FLUKA with several novel auxiliary methods. Solar modulated primary cosmic ray spectra were determined through an analysis of simultaneous proton and helium measurements made on spacecraft or high-altitude balloon flights. Primary protons and helium ions are generated within the rigidity range of 0.5 GV-20 TV, uniform in cos2theta. For a given location, primaries above the effective angle-dependent geomagnetic cut-off rigidity, and re-entrant albedo protons, are transported through the atmosphere. Helium ions are initially transported using a separate transport code called HEAVY to simulate fragmentation. HEAVY interfaces with FLUKA to provide interaction starting points for each nucleon originating from a helium nucleus. Calculated cosmic ray neutron spectra and consequent dosimetric quantities for locations with a wide range of altitude (atmospheric depth) and geomagnetic cut-off are presented and compared with measurements made on a high-altitude aeroplane. Helium ion propagation using HEAVY and inclusion of re-entrant albedo protons with the incident primary spectra significantly improved the agreement of the calculated cosmic ray neutron spectra with measured spectra. These cosmic ray propagation calculations provide the basis for a new atmospheric ionising radiation (AIR) model for air-crew dosimetry, calculation of effects on microelectronics, production of cosmogenic radionuclides and other uses. PMID- 15353686 TI - Criticality accident dosimetry systems: an international intercomparison at the SILENE reactor in 2002. AB - In criticality accident dosimetry and more generally for high dose measurements, special techniques are used to measure separately the gamma ray and neutron components of the dose. To improve these techniques and to check their dosimetry systems (physical and/or biological), a total of 60 laboratories from 29 countries (America, Europe, Asia) participated in an international intercomparaison, which took place in France from 9 to 21 June 2002, at the SILENE reactor in Valduc and at a pure gamma source in Fontenay-aux-Roses. This intercomparison was jointly organised by the IRSN and the CEA with the help of the NEA/OCDE and was partly supported by the European Communities. This paper describes the aim of this intercomparison, the techniques used by the participants and the two radiation sources and their characteristics. The experimental arrangements of the dosemeters for the irradiations in free air or on phantoms are given. Then the dosimetric quantities measured and reported by the participants are summarised, analysed and compared with the reference values. The present paper concerns only the physical dosimetry and essentially experiments performed on the SILENE facility. The results obtained with the biological dosimetry are published in two other papers of this issue. PMID- 15353687 TI - EPR dosimetry in a mixed neutron and gamma radiation field. AB - Suitability of Electron Paramagnetic Resonance (EPR) spectroscopy for criticality dosimetry was evaluated for tooth enamel, mannose and alanine pellets during the 'international intercomparison of criticality dosimetry techniques' at the SILENE reactor held in Valduc in June 2002, France. These three materials were irradiated in neutron and gamma-ray fields of various relative intensities and spectral distributions in order to evaluate their neutron sensitivity. The neutron response was found to be around 10% for tooth enamel, 45% for mannose and between 40 and 90% for alanine pellets according their type. According to the IAEA recommendations on the early estimate of criticality accident absorbed dose, analyzed results show the EPR potentiality and complementarity with regular criticality techniques. PMID- 15353688 TI - Criticality accident dosimetry by chromosomal analysis. AB - The technique of measuring the frequency of dicentric chromosomal aberrations in blood lymphocytes was used to estimate doses in a simulated criticality accident. The simulation consisted of three exposures; approximately 5 Gy with a bare source and 1 and 2 Gy with a lead-shielded source. Three laboratories made separate estimates of the doses. These were made by the iterative method of apportioning the observed dicentric frequencies between the gamma and neutron components, taking account of a given gamma/neutron dose ratio, and referring the separated dicentric frequencies to dose-response calibration curves. An alternative method, based on Bayesian ideas, was employed. This was developed for interpreting dicentric frequencies in situations where the gamma/neutron ratio is uncertain. Both methods gave very similar results. One laboratory produced dose estimates close to the eventual exercise reference doses and the other laboratories estimated slightly higher values. The main reason for the higher values was the calibration relationships for fission neutrons. PMID- 15353689 TI - A computational tool based on voxel geometry for dose reconstruction of a radiological accident due to external exposure. AB - In the case of overexposure to ionising radiation, estimation of the absorbed dose in the organism is an important indicator for evaluating the biological consequences of this exposure. The physical dosimetry approach is based either on real reconstruction of the accident, using physical phantoms, or on calculation techniques. Tools using Monte Carlo simulations associated with geometric models are very powerful since they offer the possibility to simulate faithfully the victim and the environment for dose calculations in various accidental situations. Their work presents a new computational tool, called SESAME, dedicated to dose reconstruction of radiological accidents based on anthropomorphic voxel phantoms built from real medical images of the victim in association with the MCNP Monte Carlo code. The utility was, as a first step, validated for neutrons by experimental means using a physical tissue-equivalent phantom. PMID- 15353690 TI - Czech results at criticality dosimetry intercomparison 2002. AB - Two criticality dosimetry systems were tested by Czech participants during the intercomparison held in Valduc, France, June 2002. The first consisted of the thermoluminescent detectors (TLDs) (Al-P glasses) and Si-diodes as passive neutron dosemeters. Second, it was studied to what extent the individual dosemeters used in the Czech routine personal dosimetry service can give a reliable estimation of criticality accident exposure. It was found that the first system furnishes quite reliable estimation of accidental doses. For routine individual dosimetry system, no important problems were encountered in the case of photon dosemeters (TLDs, film badge). For etched track detectors in contact with the 232Th or 235U-Al alloy, the track density saturation for the spark counting method limits the upper dose at approximately 1 Gy for neutrons with the energy >1 MeV. PMID- 15353691 TI - Reference dosimetry measurements for the international intercomparison of criticality accident dosimetry SILENE 9-21 June 2002. AB - An international intercomparison of criticality accident dosimetry systems took place in the SILENE reactor, in June 2002. Participants from 60 laboratories irradiated their dosemeters (physical and biological) using two different configurations of the reactor. In preparation for this intercomparison, the leakage radiation fields were characterised by spectrometry and dosimetry measurements using the ROSPEC spectrometer associated with a NE-213 scintillator, ionisation chambers, GM counters, diodes and thermoluminescence dosemeters (TLDs). For this intercomparison, a large area was required to irradiate the dosemeters both in free air and on phantoms. Therefore, measurements of the uniformity of the field were performed with activation detectors and TLDs for neutron and gammas, respectively. This paper describes the procedures used and the results obtained. PMID- 15353692 TI - The ENEA criticality accident dosimetry system: a contribution to the 2002 international intercomparison at the SILENE reactor. AB - The present paper summarises the activity carried out at the ENEA Radiation Protection Institute for updating the methodologies employed for the evaluation of the neutron and photon dose to the exposed workers in case of a criticality accident, in the framework of the 'International Intercomparison of Criticality Accident Dosimetry Systems' (Silene reactor, IRSN-CEA-Valduc June 2002). The evaluation of the neutron spectra and the neutron dosimetric quantities relies on activation detectors and on unfolding algorithms. Thermoluminescent detectors are employed for the gamma dose measurement. The work is aimed at accurately characterising the measurement system and, at the same time, testing the algorithms. Useful spectral information were included, based on Monte Carlo simulations, to take into account the potential accident scenarios of practical interest. All along this exercise intercomparison a particular attention was devoted to the 'traceability' of all the experimental and computational parameters and therefore, aimed at an easy treatment by the user. PMID- 15353693 TI - International intercomparison for criticality dosimetry: the case of biological dosimetry. AB - The Institute of Radiation Protection and Nuclear Safety (IRSN) organized a biological dosimetry international intercomparison with the purpose of comparing (i) dicentrics yield produced in human lymphocytes; (ii) the gamma and neutron dose estimate according to the corresponding laboratory calibration curve. The experimental reactor SILENE was used with different configurations: bare source 4 Gy, lead shield 1 and 2 Gy and a 60Co source 2 Gy. An increasing variation of dicentric yield per cell was observed between participants when there were more damages in the samples. Doses were derived from the observed dicentric rates according to the dose-effect relationship provided by each laboratory. Differences in dicentric rate values are more important than those in the corresponding dose values. The doses obtained by the participants were found to be in agreement with the given physical dose within 20%. The evaluation of the respective gamma and neutron dose was achieved only by four laboratories, with some small variations among them. PMID- 15353694 TI - Chemical dosimetry system for criticality accidents. AB - Ruder Boskovic Institute (RBI) criticality dosimetry system consists of a chemical dosimetry system for measuring the total (neutron + gamma) dose, and a thermoluminescent (TL) dosimetry system for a separate determination of the gamma ray component. The use of the chemical dosemeter solution chlorobenzene-ethanol trimethylpentane (CET) is based on the radiolytic formation of hydrochloric acid, which protonates a pH indicator, thymolsulphonphthalein. The high molar absorptivity of its red form at 552 nm is responsible for a high sensitivity of the system: doses in the range 0.2-15 Gy can be measured. The dosemeter has been designed as a glass ampoule filled with the CET solution and inserted into a pen shaped plastic holder. For dose determinations, a newly constructed optoelectronic reader has been used. The RBI team took part in the International Intercomparison of Criticality Accident Dosimetry Systems at the SILENE Reactor, Valduc, June 2002, with the CET dosimetry system. For gamma ray dose determination TLD-700 TL detectors were used. The results obtained with CET dosemeter show very good agreement with the reference values. PMID- 15353695 TI - Dose evaluation in criticality accident conditions using transient critical facilities fueled with a fissile solution. AB - Neutron dose measurement and evaluation techniques in criticality accident conditions using a thermo luminescence dosemeter (TLD) was studied at the Transient Experiment Critical Facility (TRACY) of Japan Atomic Energy Research Institute (JAERI). In the present approach, the absorbed dose is derived from the ambient dose equivalent measured with a TLD, using the appropriate conversion factor given by computation. Using this technique, the neutron dose around the SILENE reactor of the Institute for Radioprotection and Nuclear Safety (IRSN) of France was measured in the Accident Dosimetry Intercomparison Exercise (June 10 21, 2002) organized by OECD/NEA and IRSN. In this exercise, the gamma dose was also measured with a TLD. In this report, measurements and evaluation results at TRACY and SILENE are presented. PMID- 15353696 TI - A neutron dosemeter for nuclear criticality accidents. AB - A neutron dosemeter which offers instant read-out has been developed for nuclear criticality accidents. The system is based on gels containing emulsions of superheated dichlorodifluoromethane droplets, which vaporise into bubbles upon neutron irradiation. The expansion of these bubbles displaces an equivalent volume of gel into a graduated pipette, providing an immediate measure of the dose. Instant read-out is achieved using an array of transmissive optical sensors which consist of coupled LED emitters and phototransistor receivers. When the gel displaced in the pipette crosses the sensing region of the photomicrosensors, it generates a signal collected on a computer through a dedicated acquisition board. The performance of the device was tested during the 2002 International Accident Dosimetry Intercomparison in Valduc, France. The dosemeter was able to follow the initial dose gradient of a simulated accident, providing accurate values of neutron kerma; however, the emulsion was rapidly depleted of all its drops. A model of the depletion effects was developed and it indicates that an adequate dynamic range of the dose response can be achieved by using emulsions of smaller droplets. PMID- 15353697 TI - Computation of cross sections and dose conversion factors for criticality accident dosimetry. AB - In the application of criticality accident dosemeters the cross sections and fluence-to-dose conversion factors have to be computed. The cross section and fluence-to-dose conversion factor for the thermal and epi-thermal contributions to neutron dose are well documented; for higher energy regions (>100 keV) these depend on the spectrum assumed. Fluence is determined using threshold detectors. The cross sections require the folding of an expected spectrum with the reaction cross sections. The fluence-to-dose conversion factors also require a similar computation. The true and effective thresholds are used to include the information on the expected spectrum. The spectra can either be taken from compendia or measured at the facility at which the exposures are to be expected. The cross sections can be taken from data computations or analytic representations and the fluence-to-dose conversion factors are determined by various standards making bodies. The problem remaining is the method of computation. The purpose of this paper is to compare two methods for computing these factors: analytic and Monte Carlo. PMID- 15353698 TI - Response of a lithium gadolinium borate scintillator in monoenergetic neutron fields. AB - Accurate estimation of neutron dose requires knowledge of the neutron energy distribution in the working environment. Existing neutron spectrometry systems, Bonner spheres for example, are large and bulky, and require long data acquisition times. A portable system that could indicate the approximate neutron energy spectrum in a short time would be extremely useful in radiation protection. A composite scintillator, consisting of lithium gadolinium borate crystals in a plastic scintillator matrix, produced by Photogenics is being tested for this purpose. A prototype device based on this scintillator and digital pulse processing electronics has been calibrated using quasi monoenergetic neutron fields at the low-scatter facility of the UK National Physical Laboratory (NPL). Energies selected were 144, 250, 565, 1400, 2500 and 5000 keV, with correction for scattered neutrons being made using the shadow cone technique. Measurements were also made in the NPL thermal neutron field. Pulse distributions collected with the digitiser in capture-gated mode are presented, and detection efficiency and energy resolution derived. For comparison, neutron spectra were also collected using the commercially available Microspec N-Probe from Bubble Technology Industries, which consists of an NE213 scintillator and a 3He proportional counter. PMID- 15353699 TI - Characterisation of neutron fields around high-energy x-ray radiotherapy machines. AB - Photoneutron spectra around the treatment bed of a Varian Clinac 2100C machine were measured using a Bonner sphere spectrometer. To overcome problems with pulse pile-up and detection of non-neutron-induced events, the active detector of thermal neutrons normally used at the centre of the spheres was replaced by a sandwich of four CR-39 track detectors interleaved with 10B radiators. Track densities measured for the CR-39 detectors in Bonner spheres were used for the unfolding of neutron spectra. Neutron fluence and ambient dose equivalent for the whole energy range and partial energy intervals were derived from the neutron spectra. PMID- 15353700 TI - A recoil-proton spectrometer based on a p-i-n diode implementing pulse-shape discrimination. AB - A recoil-proton spectrometer was created by coupling a p-i-n diode with a polyethylene converter. The maximum detectable energy, imposed by the thickness of the totally depleted layer, is approximately 6 MeV. The minimum detectable energy is limited by the contribution of secondary electrons generated by photons in the detector assembly. This limit is approximately 1.5 MeV at full-depletion voltage and was decreased using pulse-shape discrimination. The diode was set up in the 'reverse-injection' configuration (i.e. with the N+ layer adjacent to the converter). This configuration provides longer collection times for the electron hole pairs generated by the recoil-protons. The pulse-shape discrimination was based on the zero-crossing time of bipolar signals from a (CR)2-(RC)2 filter. The detector was characterised using monoenergetic neutrons generated in the Van De Graaff CN accelerator at the INFN-Laboratori Nazionali di Legnaro. The energy limit for discrimination proved to be approximately 900 keV. PMID- 15353702 TI - Characterisation of the IRSN CANEL/T400 facility producing realistic neutron fields for calibration and test purposes. AB - The new CANEL/T400 facility has been set-up at the Institute for Radiological Protection and Nuclear Safety (IRSN) to produce a realistic neutron field. The accurate characterisation of this neutron field is mandatory since this facility will be used as a reference neutron source. For this reason an international measuring campaign, involving four laboratories with extensive expertise in neutron metrology and spectrometry, was organised through a concerted EUROMET project. Measurements were performed with Bonner sphere (BS) systems to determine the energy distribution of the emitted neutrons over the whole energy range (from thermal energy up to a few MeV). Additional measurements were performed with proton recoil detectors to provide detailed information in the energy region above 90 keV. The results obtained by the four laboratories are in agreement with each other and are compared with a calculation performed with the MCNP4C Monte Carlo code. As a conclusion of this exercise, a reliable characterisation of the CANEL/T400 neutron field is obtained. PMID- 15353701 TI - Neutron measurements in the Vandellos II nuclear power plant with a Bonner sphere system. AB - In some Spanish nuclear power plants of pressurised water reactor (PWR) type, albedo thermoluminescence dosemeters are used for personal dosimetry while survey meters, based on a thermal-neutron detector inside a cylindrical or spherical moderator, are used for dose rate assessment in routine monitoring. The response of both systems is highly dependent on the energy of the existing neutron fields. They are usually calibrated by means of ISO neutron sources with energy distributions quite different from those encountered at these installations. Spectrometric measurements with a Bonner sphere system (BSS) allow us to determine the reference dosimetric values. The UAB group, under request from the National Coordinated Research Action, was in charge of characterising the neutron fields and evaluating the response of personal dosemeters at several measurement points inside the containment building of the Catalan Nuclear Power Plant Vandellos II. The neutron fields were characterised at five places using the UAB BSS and a home made unfolding code called MITOM. The results obtained confirm the presence of low-energy components in the neutron field in most of the selected points. Moreover, we have found no influence of the nuclear fuel burning on the shape of the spectrum. PMID- 15353704 TI - A telescope-design directional neutron spectrometer. AB - A directional spectrometer that uses a superheated emulsion of dichlorotetrafluoroethane at the centre of a 30 cm diameter moderating-sphere of nylon-6. The system has a telescope-design wherein the detector views a narrow solid angle of about 1/6 steradians. The hydrogenous sphere effectively attenuates laterally incident neutrons, thus providing a strong angular dependence of the response. The central detector is sequentially operated at seven temperatures between 25 and 55 degrees C in order to generate a matrix of nested response functions suitable for few-channel spectrometry. The response matrix of the system has been determined by calibrations with monoenergetic neutrons and by Monte Carlo neutron transport calculations. The double differential unfolding method developed for this system applies the principle of maximum entropy and allows for the rigorous use of all a priori information. The spectrometer is intended for use in the mixed neutron/photon fields encountered in the nuclear power industry, being suitable for spatially distributed radiation sources with maximum neutron energies up to 10 MeV. PMID- 15353703 TI - Characterization of Bonner sphere systems at monoenergetic and thermal neutron fields. AB - The Institute for Radiological Protection and Nuclear Safety (IRSN) and the GFR, Universitat Autonoma de Barcelona (UAB) use Bonner spheres (BS) for neutron spectrometry at workplaces. The two systems, equipped with similar cylindrical 3He proportional counters, were simulated with the MCNP Monte-Carlo code to determine the response to neutrons of different energies for each polyethylene sphere. The BS systems were characterized at monoenergetic and thermal neutron fields. Measurements were performed at the Physikalisch-Technische Bundesanstalt (PTB) and at the National Physical Laboratory (NPL) standard laboratories, and with the newly characterized IRSN 'SIGMA' thermal neutron facility. The energy distribution of the reference neutron fluence was folded with the response functions for comparison purposes with the experimental data. In almost all cases related to monoenergetic neutrons, a good agreement between the experimental and the calculated count rates was found, and some discrepancies of a few per cent were observed in the thermal region. PMID- 15353705 TI - An unfolding method for directional spectrometers. AB - The development of new spectrometers that are sensitive to the directional distribution of neutrons requires new unfolding methods that can determine the distribution of the neutron fluence as a function of energy and angle. Such information is needed to compute non-isotropic dosimetric quantities (e.g. personal dose equivalent and effective dose). We describe an unfolding method that applies the maximum entropy principle to this problem. It maximises the relative entropy, defined as the information-theory entropy of the distribution of the neutron fluence relative to a distribution that encodes prior knowledge, subject to constraints imposed by the measurements. We provide examples of the applicability of the method using data from two directional spectrometers of different design that have been developed in the context of EVIDOS, a project concerning mixed neutron-photon field analysis in the nuclear industry. PMID- 15353706 TI - MITOM: a new unfolding code based on a spectra model method applied to neutron spectrometry. AB - The MITOM code was developed at UAB (Universitat Autonoma de Barcelona) for unfolding neutron spectrometric measurements with a Bonner spheres system (BSS). One of the main characteristics of this code is that an initial parameterisation of the neutron energy components (thermal, intermediate and fast) is needed. This code uses the Monte Carlo method and the Bayesian theorem to obtain a set of solutions achieving different criteria and conditions between calculated and measured count rates. The final solution is an average of the acceptable solutions. The MITOM code was tested for ISO sources and a good agreement was observed between the reference values and the unfolded ones for global magnitudes. The code was applied recently to characterise both thermal SIGMA and CANEL/T400 sources of the IRSN facilities. The results of these applications were very satisfactory as well. PMID- 15353707 TI - Multisphere default spectra--solution spectrum and dosemeter response implications. AB - Initial calibration of a multisphere spectroscopy system has been completed at Los Alamos National Laboratory using four standard calibration scenarios. Spectrum unfolding was performed using three methods of constructing the default spectrum: simple parameter models, Monte Carlo calculations and physical measurement. Comparisons of the resulting spectra for each solution method are presented. Implications of the spectral solutions upon dosemeter characterisation are addressed. PMID- 15353708 TI - Measurement and unfolding of neutron spectra using Bonner spheres. AB - Neutron spectra from Am-Be, 252Cf sources and 2H + 2H and 2H + 3H reactions have been measured using a Bonner sphere system in conjunction with a 4 x 4 mm2 LiI(Eu) detector, and unfolded using the two codes BUNKI and MAXED. The BON unfolding algorithm is used with BUNKI. It has been observed that end test percentage between 1 and 3 and smoothing factor between 0.05 and 0.1 are optimal choices for the BUNKI code. A temperature parameter 1.0 is used for MAXED. Comparison with standard spectra shows that the shape of the spectra is fairly well reproduced. A coupling between the two codes is made and the solution spectrum from BUNKI is used as the default spectrum in MAXED. PMID- 15353709 TI - In phantom Dosimetric response of tooth enamel to neutrons. AB - Electron Paramagnetic Resonance dosimetry based on tooth enamel has one important application in dose reconstruction of nuclear plant workers, where the contribution of neutrons to individual dose is often important. Evaluation of tooth enamel response to neutrons is thus an important goal. A few experimental data at thermal and fast neutron energies are available. A first evaluation of the tooth enamel relative response to 60Co in monoenergetic neutron flux of 2.8 and of 14 MeV, published elsewhere, has provided results apparently non consistent with the results obtained at lower and higher energies. A comparison of those results in the 2.8 and 14 MeV beams with those available in the literature for other beams is reported and possible reasons for incongruities are discussed. Dose conversion factors of enamel to the water and air are also calculated and reported. PMID- 15353710 TI - Dosimetry of clinical neutron and proton beams: an overview of recommendations. AB - Neutron therapy beams are obtained by accelerating protons or deuterons on Beryllium. These neutron therapy beams present comparable dosimetric characteristics as those for photon beams obtained with linear accelerators; for instance, the penetration of a p(65)+Be neutron beam is comparable with the penetration of an 8 MV photon beam. In order to be competitive with conventional photon beam therapy, the dosimetric characteristics of the neutron beam should therefore not deviate too much from the photon beam characteristics. This paper presents a brief summary of the neutron beams used in radiotherapy. The dosimetry of the clinical neutron beams is described. Finally, recent and future developments in the field of physics for neutron therapy is mentioned. In the last two decades, a considerable number of centres have established radiotherapy treatment facilities using proton beams with energies between 50 and 250 MeV. Clinical applications require a relatively uniform dose to be delivered to the volume to be treated, and for this purpose the proton beam has to be spread out, both laterally and in depth. The technique is called 'beam modulation' and creates a region of high dose uniformity referred to as the 'spread-out Bragg peak'. Meanwhile, reference dosimetry in these beams had to catch up with photon and electron beams for which a much longer tradition of dosimetry exists. Proton beam dosimetry can be performed using different types of dosemeters, such as calorimeters, Faraday cups, track detectors and ionisation chambers. National standard dosimetry laboratories will, however, not provide a standard for the dosimetry of proton beams. To achieve uniformity on an international level, the use of an ionisation chamber should be considered. This paper reviews and summarises the basic principles and recommendations for the absorbed dose determination in a proton beam, utilising ionisation chambers calibrated in terms of absorbed dose to water. These recommendations are based on the recent IAEA TRS398 Code of Practice: 'Absorbed Dose Determination in External Beam Radiotherapy: An International Code of Practice for Dosimetry based on Standards of Absorbed Dose to Water'. PMID- 15353711 TI - Application of semiconductors for dosimetry of fast-neutron therapy beam. AB - Two types of ion implanted miniature p-i-n diodes were tested in a d(48.5) + Be fast-neutron beam produced in the Detroit superconducting cyclotron. The increase in forward voltage drop caused by neutron-induced damage was correlated with neutron dose measured in a water phantom. The neutron and gamma dose components were predetermined using twin detector (Tissue-equivalent ion chamber paired with miniature Geiger-Muller counter) method. The increase in the voltage drop for 1 mA injection current was monitored together with the cyclotron beam target current, thus the differential voltage drop could be defined precisely for given radiation dose. The average neutron sensitivities of tested diodes were 1.284 +/- 0.014 and 0.528 +/- 0.058 mV per cGy. The miniature detectors can be utilised in characterisation of small radiation fields and in the regions of high dose gradient as well as for in vivo dosimetry of the patients undergoing fast-neutron therapy. PMID- 15353712 TI - BNCT microdosimetry at the tapiro reactor thermal column. AB - A thermal column is available for dosimetric and radiobiological studies by the fast reactor TAPIRO, located at the ENEA research centre Casaccia. The TAPIRO neutron field has been studied (in the frame of LNL BNCT project) with a tissue equivalent proportional counter, which has worked alternatively with an ordinary tissue-equivalent cathode and with a boron-enriched cathode. Measurements have been performed with polyethylene caps of different thickness. Both the absorbed dose and the microdosimetric-calculated biological effective dose show a maximum at approximately 0.5 mg cm(-2) of depth. The different dose components have been calculated and the results are discussed. PMID- 15353713 TI - A toolkit for epithermal neutron beam characterisation in BNCT. AB - Methods for dosimetry of epithermal neutron beams used in boron neutron capture therapy (BNCT) have been developed and utilised within the Finnish BNCT project as well as within a European project for a code of practise for the dosimetry of BNCT. One outcome has been a travelling toolkit for BNCT dosimetry. It consists of activation detectors and ionisation chambers. The free-beam neutron spectrum is measured with a set of activation foils of different isotopes irradiated both in a Cd-capsule and without it. Neutron flux (thermal and epithermal) distribution in phantoms is measured using activation of Mn and Au foils, and Cu wire. Ionisation chamber (IC) measurements are performed both in-free-beam and in phantom for determination of the neutron and gamma dose components. This toolkit has also been used at other BNCT facilities in Europe, the USA, Argentina and Japan. PMID- 15353714 TI - Fast neutrons produced by nuclear fragmentation in treatment irradiations with 12C beam. AB - In the framework of the heavy-ion tumour therapy project at GSI we investigated the nuclear fragmentation of 200 AMeV carbon ions stopping in a 12.78-cm thick water absorber. Fast neutrons and charged particles emerging from the target were registered at forward angles between 0 degrees and 30 degrees with a DeltaE-E telescope consisting of an NE102 and a BaF2 scintillator. We obtained neutron energy spectra and angular distributions and derived the neutron yield in the energy range from 10 to 500 MeV in the forward hemisphere. In addition, we performed fragmentation measurements in actual patient treatment irradiations. The resulting angular distributions of neutrons and charged particles as well as their yields are similar to those obtained with the water absorber. PMID- 15353715 TI - Neutron field produced by 25 MeV deuteron on thick beryllium for radiobiological study; energy spectrum. AB - Biological data is necessary for estimation of protection from neutrons, but there is a lack of data on biological effects of neutrons for radiation protection. Radiological study on fast neutrons has been done at the National Institute of Radiological Sciences. An intense neutron source has been produced by 25 MeV deuterons on a thick beryllium target. The neutron energy spectrum, which is essential for neutron energy deposition calculation, was measured from thermal to maximum energy range by using an organic liquid scintillator and multi sphere moderated 3He proportional counters. The spectrum of the gamma rays accompanying the neutron beam was measured simultaneously with the neutron spectrum using the organic liquid scintillator. The transmission by the shield of the spurious neutrons originating from the target was measured to be less than 1% by using the organic liquid scintillator placed behind the collimator. The measured neutron energy spectrum is useful in dose calculations for radiobiology studies. PMID- 15353716 TI - Peripheral neutron and gamma doses in radiotherapy with an 18 MV linear accelerator. AB - More and more attention is being given in radiotherapy to the doses received by organs other than the target organ. With increasing survival time of the patients, the risks of secondary malignancies need to be lowered as much as possible. So total body doses are worth estimating in radiotherapy. The introduction of intensity modulated radiotherapy (IMRT) needs an increase in the number of monitor units given to the patient. So there is a risk of increasing the peripheral doses using this technique. Another aspect, mostly neglected, is the neutron peripheral dose that occurs when LINAC energies above 8 MeV are used. We did measurements for both gammas and neutrons with an 18 MV Varian accelerator for a prostate cancer treatment. The measurements were done both free-in-air, at different depths in a plexi-phantom, and using a Rando-Alderson phantom. Effective doses for the total body outside the treatment area are estimated using these measurements. PMID- 15353717 TI - Thermoluminescence dosimetry in mixed neutron/gamma radiation beam. AB - Miniature CaF2:Tm (TLD-300) were adopted for dosimetry in a mixed neutron/gamma beam produced by the Detroit superconducting cyclotron. A method utilizing the linear relationship between the ratios of the areas under the low and high temperature peaks of the glow curve and the ratios of gamma dose to total dose (Dgamma/DT) in d(48.5)+Be fast-neutron beam was applied for both the gamma component and the total dose measurements. The detectors were grouped based on their response to uniform 60Co dose. Within each group the ratio of the peak areas of the glow curve demonstrated uniformity and stability better than 1.5% (one standard deviation). The parameters of the linear regression between the fractional gamma component in the neutron beam and the ratio of peak areas were derived from measurements at locations with predetermined Dgamma/DT. The method was applied to measure the transmission through the existing multi-rod and newly designed multi-leaf collimators. PMID- 15353718 TI - Development of a tiny neutron probe with an optical fibre for BNCT. AB - We have developed a tiny neutron probe detector as a monitor of a thermal neutron flux for boron neutron capture therapy. The detector consists of an optical fibre and a small neutron probe. We have used a film-like ZnS(Ag) scintillator and a 6LiF neutron converter for the neutron probe. In order to improve the gamma neutron discrimination ability, vacuum evaporation of 6LiF onto the ZnS(Ag) film has been done. In order to improve the neutron detection efficiency, we made use of a wavelength-shifting fibre as the probe material. The characteristics of the above two types of fibre probe detector have been evaluated experimentally. PMID- 15353719 TI - Dosimetry of BNCT beams with novel thermoluminescent detectors. AB - In the dosimetry of boron neutron capture therapy (BNCT) beams, thermoluminescent (TL) detectors are typically applied in phantom measurements to determine the spatial distribution of the gamma ray and neutron dose. Pairs of 6LiF and 7LiF are applied to discriminate between the thermal neutron and gamma ray field components, exploiting the high cross section for (n,alpha) reaction of 6Li. At the Institute of Nuclear Physics (INP) in Krakow (Poland) a prototype TL-based measuring set has been constructed and tested. This set consists of a miniature TL detector (of 2 mm diameter and 0.4 mm thickness) placed inside a miniature container made of non-thermoluminescent 6LiF. The outer dimensions of the set are 4.5 mm diameter and 1.4 mm thickness, enabling its application in place of a thermoluminescence dosemeter pellet in typical phantoms. The detector sets were tested in the BNCT beam of the Studsvik reactor. By exploiting the ratio of TL signals of the unshielded and shielded detectors, it was possible to estimate the contributions of the thermal and epithermal components of the neutron field. PMID- 15353720 TI - Combined TL and 10B-alanine ESR dosimetry for BNCT. AB - The dosimetric technique described in this paper is based on electron spin resonance (ESR) detectors using an alanine-boric compound acid enriched with (10)B, and beryllium oxide thermoluminescent (TL) detectors; with this combined dosimetry, it is possible to discriminate the doses due to thermal neutrons and gamma radiation in a mixed field. Irradiations were carried out inside the thermal column of a TRIGA MARK II water-pool-type research nuclear reactor, also used for Boron Neutron Capture therapy (BNCT) applications, with thermal neutron fluence from 10(9) to 10(14) nth cm(-2). The ESR dosemeters using the alanine boron compound indicated ESR signals about 30-fold stronger than those using only alanine. Moreover, a negligible correction for the gamma contribution, measured with TL detectors, almost insensitive to thermal neutrons, was necessary. Therefore, a simultaneous analysis of our TL and ESR detectors allows discrimination between thermal neutron and gamma doses, as required in BNCT. PMID- 15353721 TI - Study of a method based on TLD detectors for in-phantom dosimetry in BNCT. AB - A method has been developed, based on thermoluminescent dosemeters (TLD), aimed at measuring the absorbed dose in tissue-equivalent phantoms exposed to thermal or epithermal neutrons, separating the contributions of various secondary radiation generated by neutrons. The proposed method takes advantage of the very low sensitivity of CaF2:Tm (TLD-300) to low energy neutrons and to the different responses to thermal neutrons of LiF:Mg,Ti dosemeters with different 6Li percentage (TLD-100, TLD-700, TLD-600). The comparison of the results with those obtained by means of gel dosemeters and activation foils has confirmed the reliability of the method. The experimental modalities allowing reliable results have been studied. The glow curves of TLD-300 after gamma or neutron irradiation have been compared; moreover, both internal irradiation effect and energy dependence have been investigated. For TLD-600, TLD-100 and TLD-700, the suitable fluence limits have been determined in order to avoid radiation damage and loss of linearity. PMID- 15353722 TI - Alanine and TLD coupled detectors for fast neutron dose measurements in neutron capture therapy (NCT). AB - A method was investigated to measure gamma and fast neutron doses in phantoms exposed to an epithermal neutron beam designed for neutron capture therapy (NCT). The gamma dose component was measured by TLD-300 [CaF2:Tm] and the fast neutron dose, mainly due to elastic scattering with hydrogen nuclei, was measured by alanine dosemeters [CH3CH(NH2)COOH]. The gamma and fast neutron doses deposited in alanine dosemeters are very near to those released in tissue, because of the alanine tissue equivalence. Couples of TLD-300 and alanine dosemeters were irradiated in phantoms positioned in the epithermal column of the Tapiro reactor (ENEA-Casaccia RC). The dosemeter response depends on the linear energy transfer (LET) of radiation, hence the precision and reliability of the fast neutron dose values obtained with the proposed method have been investigated. Results showed that the combination of alanine and TLD detectors is a promising method to separate gamma dose and fast neutron dose in NCT. PMID- 15353723 TI - Microdosimetry of neutron field for boron neutron capture therapy at Kyoto university reactor. AB - Microdosimetric single event spectrum in a human body simulated by an acrylic phantom has been measured for the clinical BNCT field at the Kyoto University Reactor (KUR). The recoil particles resulting from the initial reaction and subsequent interactions, namely protons, electrons, alpha particles and carbon nuclei are identified in the microdosimetric spectrum. The relative contributions to the neutron dose from proton, alpha particles and carbon are estimated to be about 0.9, 0.07 and 0.3, respectively, four depths between 5 and 41 mm. We estimate that the dose averaged lineal energy, yD decreased with depth from 64 to 46 keV microm(-1). Relative biological effectiveness (RBE) of this neutron field using a response function for the microdosimetric spectrum was estimated to decrease from 3.6 to 2.9 with increasing depth. PMID- 15353724 TI - Characterisation of the TAPIRO BNCT epithermal facility. AB - A collimated epithermal beam for boron neutron capture therapy (BNCT) research has been designed and built at the TAPIRO fast research reactor. A complete experimental characterisation of the radiation field in the irradiation chamber has been performed, to verify agreement with IAEA requirements. Slow neutron fluxes have been measured by means of an activation technique and with thermoluminescent detectors (TLDs). The fast neutron dose has been determined with gel dosemeters, while the fast neutron spectrum has been acquired by means of a neutron spectrometer based on superheated drop detectors. The gamma-dose has been measured with gel dosemeters and TLDs. For an independent verification of the experimental results, fluxes, doses and neutron spectra have been calculated with Monte Carlo simulations using the codes MCNP4B and MCNPX_2.1.5 with the direct statistical approach (DSA). The results obtained confirm that the epithermal beams achievable at TAPIRO are of suitable quality for BNCT purposes. PMID- 15353725 TI - Characterisation of the TAPIRO BNCT thermal facility. AB - Dosimetry and spectrometry measurements have been carried out in the thermal column of the research fast reactor RSV-TAPIRO (ENEA-Casaccia, Rome) in order to investigate its suitability for irradiation of cells or mice, with a view to research in the interests of boron neutron capture therapy (BNCT). The thermal column consists of a graphite moderator (40 cm thick) containing a lead shield (13 cm thick) in order to shield reactor background. The irradiation volume, inside this structure, has cubic shape (18 x 18 x 18 cm3). Besides measurements of fluence and dose rates in air or in phantom performed with thermoluminescence dosemeters (TLDs) and using the activation technique, dose and fluence profiles have been generated using a method based on gel dosemeters analysed with optical imaging. To check the consistency of the results, spectrometry measurements in the same irradiation volume have been performed by means of bubble detectors. PMID- 15353726 TI - An accelerator-based epithermal neutron beam design for BNCT and dosimetric evaluation using a voxel head phantom. AB - The beam shaping assembly design has been investigated in order to improve the epithermal neutron beam for accelerator-based boron neutron capture therapy in intensity and quality, and dosimetric evaluation for the beams has been performed using both mathematical and voxel head phantoms with MCNP runs. The neutron source was assumed to be produced from a conventional 2.5 MeV proton accelerator with a thick (7)Li target. The results indicate that it is possible to enhance epithermal neutron flux remarkably as well as to embody a good spectrum shaping to epithermal neutrons only with the proper combination of moderator and reflector. It is also found that a larger number of thermal neutrons can reach deeply into the brain and, therefore, can reduce considerably the treatment time for brain tumours. Consequently, the epithermal neutron beams designed in this study can treat more effectively deep-seated brain tumours. PMID- 15353727 TI - Use of the CT images for BNCT calculation: development of BNCT treatment planning system and its applications to dose calculation for voxel phantoms. AB - A BNCT (Boron Neutron Capture Therapy) treatment planning system (BTPS) was developed for BNCT study and treatment planning. Three kinds of CT images, VHP, PINNACLE and DICOM images, were employed to make voxel phantoms for BNCT patient treatment using the BTPS. The thermal neutron, fast neutron, gamma and boron doses are calculated and background, tissue, and tumour doses for idealised standard reactor neutron field (ISRNF) neutron beam were calculated by using BTPS and MCNP code. It was noted that the total computing times needed for BNCT analysis could be greatly reduced since the BTPS system provides a dose analysis tool and a lengthy MCNP input in a short time. It is, thus, expected that the BTPS can significantly contribute the BNCT study for the treatment of patients. PMID- 15353728 TI - Recombination chambers filled with different gases--studies of possible application for BNCT beam dosimetry. AB - Recombination microdosimetric method (RMM), based on the phenomenon of initial recombination of ions is applied to determine the distribution of the absorbed dose versus linear energy transfer (LET). Usually, the recombination chambers used for RMM are filled with tissue-equivalent gas, but the response of the device can be adjusted to the actual needs by the use of different gases. Using a graphite chamber filled with nitrogen and 10BF3 it was shown that RMM can also be used with chambers containing these gases. This opens the possibility of designing a recombination chamber for the determination of the dose fractions due to gamma radiation, fast neutrons, neutron capture on nitrogen and high-LET particles from the (n,10B) reaction in simulated tissue with different contents of 10B. It was also necessary to improve the method for the determination of initial recombination at low polarising voltages, when volume-recombination and back-diffusion of ions are considerably high. PMID- 15353729 TI - Passive spectrometry of linear energy transfer: development and use. AB - A linear energy transfer (LET) spectrometer based on the evaluation of particle track parameters in a chemically etched polyallyldiglycolcarbonate (PADC) track detector has been developed at our laboratory. It permits us to determine LET spectra between 10 and 700 keV microm(-1) in tissues. The LET spectra obtained permit us to calculate total dose and dose equivalent corresponding to particles with etchable tracks also. We have recently been able to verify the calibration curves used by using C, Mg, Ne, Si and Fe ion beams with different energies. The calibration curves obtained are presented and compared with those originally used, and a good correlation is found. The LET spectrometer with new calibration was used to analyse the radiation quality of the radiotherapy proton beam at the Joint Institute for Nuclear Research (JINR). The radiation quality was studied along the proton's range, particular attention being devoted to the region of the Bragg peak. It was found that the biologically weighted effective dose (BWE) reaches a value of about 1.25 at the Bragg peak region. At the beam entrance this value increases to about 1.02 due to secondary particles created through primary proton nuclear reactions in tissues. PMID- 15353730 TI - Microdosimetric investigation at the therapeutic proton beam facility of CATANA. AB - Proton beams (62 Mev) are used by the Laboratori Nazionali del Sud of the Italian Institute of Nuclear Physics to treat eye melanoma tumours at the therapeutic facility called CATANA. A cylindrical slim tissue-equivalent proportional counter (TEPC) of 2.7 mm external diameter has been used to compare the radiation quality of two spread-out Bragg peaks (SOBP) at the CATANA proton beam. PMID- 15353731 TI - Dosimetric measurements with a brain equivalent plastic walled ionization chamber in an epithermal neutron beam. AB - The tissue substitute A-181 plastic, which has an elemental composition matching both the constituent hydrogen and nitrogen of brain tissue, was assessed for dosimetry in boron neutron capture therapy (BNCT). The sensitivity of an A-181 walled ionization chamber relative to photons for all neutrons in a clinical epithermal beam was calculated to vary between 0.79 +/- 0.04 in-air and 0.95 +/- 0.01 at depths of 4 cm and greater in-phantom. Differences in the total neutron doses measured with A-150 and A-181 plastic-walled chambers were attributed, within experimental error, to the dose produced by thermal neutron capture reactions from the different concentrations of nitrogen in the two tissue substitutes. The response of the A-181 chamber was converted to total neutron dose with an uncertainty increasing with depth in-phantom from 13 to 23% the magnitude of which is determined by the subtraction of a relatively large photon dose. The use of A-181 in place of A-150 plastic will no longer require partitioning the measured neutron dose by energy and should simplify dose reporting in BNCT. PMID- 15353732 TI - Neutron fluence-to-dose conversion coefficients for embryo and fetus. AB - A problem of concern in radiation protection is the exposure of pregnant women to ionising radiation, because of the high radiosensitivity of the embryo and fetus. External neutron exposure is of concern when pregnant women travel by aeroplane. Dose assessments for neutrons frequently rely on fluence-to-dose conversion coefficients. While neutron fluence-to-dose conversion coefficients for adults are recommended in International Commission on Radiological Protection publications and International Commission on Radiological Units and Measurements reports, conversion coefficients for embryos and fetuses are not given in the publications. This study undertakes Monte Carlo calculations to determine the mean absorbed doses to the embryo and fetus when the mother is exposed to neutron fields. A new set of mathematical models for the embryo and fetus has been developed at Health Canada and is used together with mathematical phantoms of a pregnant female developed at Oak Ridge National Laboratory. Monoenergetic neutrons from 1 eV to 10 MeV are considered in this study. The irradiation geometries include antero-posterior (AP), postero-anterior (PA), lateral (LAT), rotational (ROT) and isotropic (ISO) geometries. At each of these standard irradiation geometries, absorbed doses to the fetal brain and body are calculated; for the embryo at 8 weeks and the fetus at 3, 6 or 9 months. Neutron fluence-to-absorbed dose conversion coefficients are derived for the four age groups. Neutron fluence-to-equivalent dose conversion coefficients are given for the AP irradiations which yield the highest radiation dose to the fetal body in the neutron energy range considered here. The results indicate that for neutrons <10 MeV more protection should be given to pregnant women in the first trimester due to the higher absorbed dose per unit neutron fluence to the fetus. PMID- 15353733 TI - Using operational equipment to read accident dosemeters. AB - Analysis of accident dosemeters usually involves the use of laboratory-based counting equipment. Gamma spectrometers are used for indium, copper and gold, and alpha-beta detectors for sulphur. This equipment is usually not easily transported due to the shielding required and the weight and delicacy of the counters. For intercomparison studies that require reading the dosemeters on site, a transportable system is required unless the site operating the study can count samples for all the participants. In the case of an actual accident these systems would have a difficulty in counting a large number of accident dosemeters. In an accident, personnel are usually subdivided according to their level of exposure. Those exposed to higher doses are treated immediately. An alternate system should be made available to handle the dosemeters worn by those personnel are likely to receive lower doses. Improvements in portable operational equipment for gamma and beta monitoring allow their use as spectrometers. Such a system was used for the SILENE intercomparison conducted at IRSN Valduc on 12 June and 19, 2002, and the preliminary results compared well with the other participants. PMID- 15353734 TI - Personal neutron dosimetry in nuclear power plants using etched track and albedo thermoluminescence dosemeters. AB - Measurement of the personal dose equivalent rates for neutrons is a difficult task because available dosemeters do not provide the required energy response and sensitivity. Furthermore, the available wide calibration spectra recommended by the International Standard Organisation does not reproduce adequately the spectra encountered in practical situations of the nuclear industry. There is a real necessity to characterise the radiation field, in which workers can be exposed, and to calibrate personal dosemeters in order to determine the dose equivalent in these installations. For this reason, we measure the neutron spectrum with our Bonner sphere system and we fold this spectrum with energy-dependent fluence-to dose conversion coefficients to obtain the reference dose equivalent rate. This reference value is then compared with the personal dosemeter reading to determine a field-specific correction factor. In this paper, we present the values of this field-specific correction factor for etched track and albedo thermoluminescence dosemeters at three measurement locations inside the containment building of the Vandellos II nuclear power plant. We have found that assigning to each personal dosemeter the mean value of the field-specific correction factors of the three measurement locations, allows the evaluation of neutron personal dose equivalent rate with a relative uncertainty of approximately 25 and 15% for the PADC and albedo dosemeters, respectively. PMID- 15353735 TI - Calibration and testing of a TLD dosemeter for area monitoring. AB - The response of a TLD-600/TLD-700 area dosemeter has been characterized in neutron fields around the 590 MeV cyclotron ring at the Paul Scherrer Institute (PSI). The dosemeter is based on a cylindrical paraffin moderator with three of each type of TLD chip at the centre, and is intended to use for area monitoring around accelerator facilities. The dosemeter is calibrated in terms of ambient dose equivalent using a non-moderated 252Cf neutron source. The ambient dose equivalent response has been tested in five locations where the neutron fields and dose rates have been well characterized by Bonner sphere spectrometer and active neutron monitor measurements. The different spectrum shapes and dose rates in the five locations permit the comparison of the behavior of the active and passive dosemeters in these neutron fields. PMID- 15353736 TI - Neutron field measurements for alara purposes around a Van de Graaff accelerator building. AB - The Institute for Reference Materials and Measurements operates a 7.0 MV Van de Graaff accelerator to generate monoenergetic neutron radiation for experimental applications. Owing to increased intensities of generated neutron fields and the more stringent regulation related to the maximum dose for the public, a concrete shielding wall surrounding the experimental building was constructed. This paper presents a study aiming at evaluating the effect of the shielding on the neutron field outside the wall. For this purpose, the following measurements were carried out around the building: (1) cartography of the neutron field for different experimental conditions; (2) measurement of neutron spectra using multiple Bonner spheres; (3) activation measurements using gold discs followed by low-level gamma spectrometry. From the measurements, it can be concluded that the wall fulfils its purpose to reduce the neutron dose rate to the surrounding area to an acceptable level. PMID- 15353737 TI - Measurement of the neutron fluence and dose spectra using an extended bonner sphere and a tissue-equivalent proportional counter. AB - A conventional Bonner Sphere (BS) set consisting of six polyethylene spheres was modified to enhance its response to a high-energy neutron by putting a lead shell inside a polyethylene moderator. The response matrix of an extended BS was calculated using the MCNPX code and calibrated using a 252Cf neutron source. In order to survey the unknown photon and neutron mixed field, a spherical tissue equivalent proportional counter (TEPC) was constructed and assembled as a portable measurement system. The extended BS and the self-constructed TEPC were employed to determine the dosimetric quantities of the neutron field produced from the thick lead target bombarded by the 2.5 GeV electron beam of Pohang Accelerator Laboratory (PAL) and the neutron calibration field of Korea Atomic Energy Research Institute (KAERI). PMID- 15353738 TI - Methodology for worker neutron exposure evaluation in the PDCF facility design. AB - A project headed by Washington Group International is meant to design the Pit Disassembly and Conversion Facility (PDCF) to convert the plutonium pits from excessed nuclear weapons into plutonium oxide for ultimate disposition. Battelle staff are performing the shielding calculations that will determine appropriate shielding so that the facility workers will not exceed target exposure levels. The target exposure levels for workers in the facility are 5 mSv y(-1) for the whole body and 100 mSv y(-1) for the extremity, which presents a significant challenge to the designers of a facility that will process tons of radioactive material. The design effort depended on shielding calculations to determine appropriate thickness and composition for glove box walls, and concrete wall thicknesses for storage vaults. Pacific Northwest National Laboratory (PNNL) staff used ORIGEN-S and SOURCES to generate gamma and neutron source terms, and Monte Carlo (computer code for) neutron photon (transport) (MCNP-4C) to calculate the radiation transport in the facility. The shielding calculations were performed by a team of four scientists, so it was necessary to develop a consistent methodology. There was also a requirement for the study to be cost effective, so efficient methods of evaluation were required. The calculations were subject to rigorous scrutiny by internal and external reviewers, so acceptability was a major feature of the methodology. Some of the issues addressed in the development of the methodology included selecting appropriate dose factors, developing a method for handling extremity doses, adopting an efficient method for evaluating effective dose equivalent in a non-uniform radiation field, modelling the reinforcing steel in concrete, and modularising the geometry descriptions for efficiency. The relative importance of the neutron dose equivalent compared with the gamma dose equivalent varied substantially depending on the specific shielding conditions and lessons were learned from this effect. This paper addresses these issues and the resulting methodology. PMID- 15353739 TI - Propagation characteristics of neutrons leaking from the accelerator facilities. AB - In this study spatial and time distribution of neutrons leaking from Hokkaido University 45 MeV Electron Linac facility have measured and compared with the Monte Carlo simulations. The neutron transport processes inside and outside the facility building has been simulated using MCNP. The neutrons have measured by BF3 counters and 3He counters with polyethylene moderators up to the distance of 330 m from the facility. The spatial distribution of ambient dose equivalent converted from the counts has been compared with the simulation. The distribution estimated from the counts by the BF3 counter has been shown fairly good agreement with the calculation. The spatial distribution of counts obtained at the 45 MeV Electron Linac facility has been compared with that obtained at the Fusion Neutronics Source (FNS) facility of JAERI. The difference between the propagation characteristics of neutrons leaking from those facilities has been discussed. PMID- 15353740 TI - Neutron dose distribution at the GSI fragment separator. AB - GSI is operating a facility for the production of rare isotopes. Nuclei are produced by fragmentation or fission of the impinging heavy ions with energies of approximately 1 GeV per nucleon. The major part of the primary beam and the produced nuclei is deposited in the components of the Fragment Separator (FRS) and generates neutron radiation. Thermoluminescence dosemeters (TLDs) (6LiF/7LiF pairs in PE spheres) were exposed in neutron fields produced by uranium beams with energies between 100 and 1000 MeV per nucleon during an irradiation period in the year 2002. Two-dimensional dose distributions are obtained using these TL measurements in combination with model calculations. The applied model describes the dose distribution as a superposition of dose patterns of 20 single sources equally distributed along the FRS. The single source distribution is based on a measured double differential neutron distribution for a 1 GeV per nucleon uranium beam. PMID- 15353741 TI - Current problems and expected improvements in personal neutron dosimetry. AB - Many technological activities involve the potential for worker exposure to neutrons. The determination of neutron personal dose equivalent is difficult due to a number of factors including the materials and methods used to evaluate the response of personal dosemeters and the quantities for expressing dose equivalent. Nevertheless, recent progress has been made in the development of devices and techniques for the measurement and calibration of neutron personal dosemeters. The quantities and units used to express neutron dose equivalent are being improved and clarified. Therefore, it is expected that a number of remaining difficulties with neutron dosimetry will be mitigated. PMID- 15353742 TI - Electronic personal dosemeters: the solution to problems of individual monitoring in mixed neutron/photon fields? AB - An overview is given showing the main principles of the present-day electronic neutron dosemeters. The radiological performance of the devices is described in a comparative way. This includes chiefly the personal dose equivalent Hp(10) response for monoenergetic neutrons and in practical fields with broad energy distributions and estimations of the low dose limit for neutrons. PMID- 15353743 TI - Individual neutron monitoring in workplaces with mixed neutron/photon radiation. AB - EVIDOS ('evaluation of individual dosimetry in mixed neutron and photon radiation fields') is an European Commission (EC)-sponsored project that aims at a significant improvement of radiation protection dosimetry in mixed neutron/photon fields via spectrometric and dosimetric investigations in representative workplaces of the nuclear industry. In particular, new spectrometry methods are developed that provide the energy and direction distribution of the neutron fluence from which the reference dosimetric quantities are derived and compared to the readings of dosemeters. The final results of the project will be a comprehensive set of spectrometric and dosimetric data for the workplaces and an analysis of the performance of dosemeters, including novel electronic dosemeters. This paper gives an overview of the project and focuses on the results from measurements performed in calibration fields with broad energy distributions (simulated workplace fields) and on the first results from workplaces in the nuclear industry, inside a boiling water reactor and around a spent fuel transport cask. PMID- 15353744 TI - Dose equivalent measurements in a strongly pulsed high-energy radiation field. AB - The stray radiation field outside the shielding of high-energy accelerators comprises neutrons, photons and charged particles with a wide range of energies. Often, accelerators operate by accelerating and ejecting short pulses of particles, creating an analogue, pulsed radiation field. The pulses can be as short as 10 micros with high instantaneous fluence rates and dose rates. Measurements of average dose equivalent (rate) for radiation protection purposes in these fields present a challenge for instrumentation. The performance of three instruments (i.e. a recombination chamber, the Sievert Instrument and a HANDI TEPC) measuring total dose equivalent is compared in a high-energy reference radiation field (CERF) and a strongly pulsed, high-energy radiation field at the CERN proton synchrotron (PS). PMID- 15353745 TI - Practical implications of neutron survey instrument performance. AB - Improvements have been made to the Monte Carlo modelling used to calculate the response of the neutron survey instruments most commonly used in the UK, for neutron energies up to 20 MeV. The improved modelling of the devices includes the electronics and battery pack, allowing better calculations of both the energy and angle dependence of response. These data are used to calculate the response of the instruments in rotationally and fully isotropic, as well as unidirectional fields. Experimental measurements with radionuclide sources and monoenergetic neutron fields have been, and continue to be made, to test the calculated response characteristics. The enhancements to the calculations have involved simulation of the sensitivity of the response to variations in instrument manufacture, and will include the influence of the user and floor during measurements. The practical implications of the energy and angle dependence of response, variations in manufacture, and the influence of the user are assessed by folding the response characteristics with workplace energy and direction distributions. PMID- 15353746 TI - Intercomparison on the usage of computational codes in radiation dosimetry. AB - 'QUADOS', a Concerted Action of the European Commission, has run an intercomparison aimed at evaluating the use of computational codes for dosimetry in radiation protection and medical physics. This intercomparison was open to all users of Monte Carlo, analytic and semi-analytic codes or deterministic methods. Its main aim was to provide a snapshot of the methods and codes currently in use. It also intended to furnish information on the methods used to assess the reliability of computational results and disseminate 'good practice' throughout the radiation dosimetry community. Eight problems were selected for their relevance to the radiation dosimetry community, three of which involve neutron transport. This paper focuses on the analysis of the neutron problems. PMID- 15353747 TI - Weighting factors for radiation quality: how to unite the two current concepts. AB - The quality factor, Q(L), used to be the universal weighting factor to account for radiation quality, until--in its 1991 Recommendations--the ICRP established a dichotomy between 'computable' and 'measurable' quantities. The new concept of the radiation weighting factor, w(R), was introduced for use with the 'computable' quantities, such as the effective dose, E. At the same time, the application of Q(L) was restricted to 'measurable' quantities, such as the operational quantities ambient dose equivalent or personal dose equivalent. The result has been a dual system of incoherent dosimetric quantities. The most conspicuous inconsistency resulted for neutrons, for which the new concept of wR had been primarily designed. While its definition requires an accounting for the gamma rays produced by neutron capture in the human body, this effect is not adequately reflected in the numerical values of wR, which are now suitable for mice, but are--at energies of the incident neutrons below 1 MeV--conspicuously too large for man. A recent Report 92 to ICRP has developed a proposal to correct the current imbalance and to define a linkage between the concepts Q(L) and wR. The proposal is here considered within a broader assessment of the rationale that led to the current dual system of dosimetric quantities. PMID- 15353748 TI - Recent advances of nanodosimetry. AB - The early damage to genes and cells due to ionizing radiation is initiated by the overlay of the track structure of charged particles and of the structure of radiosensitive sub-cellular volumes. As a result of this overlay, a specified number of ionizations (the ionization cluster size) is formed per primary particle. Therefore, one of the aims of nanodosimetry is to determine ionization cluster-size distributions in nanometric volumes of liquid water, as a substitute to sub-cellular structures. After a short description of the main aspects of cluster-size formation by charged particles, an overview of the advanced measuring techniques that use millimetric target volumes filled with a low pressure gas to simulate nanometric target volumes at unit density is given. Afterwards, physical principles are discussed which are applicable to convert ionization cluster-size distributions measured in gases into those for liquid water. Finally, a tentative possibility is proposed of how to relate parameters derived from cluster-size distributions in liquid water to parameters derived from radiation-induced radiobiological experiments. PMID- 15353749 TI - Compound dual radiation action theory for 252Cf brachytherapy. AB - The existing dosimetry protocol that uses the concept of RBE for 252Cf brachytherapy contains large uncertainties. A new formula has been developed to correlate the biological effect (i.e. cell survival fraction) resulting from a mixed n + gamma radiation field with two physical quantities and two biological quantities. The formula is based on a pathway model evolved from that of the compound-dual-radiation-action (CDRA) theory, previously proposed by Rossi and Zaider. The new model employs the recently published data on radiation-induced DNA lesions. The new formula is capable of predicting quantitatively the synergistic effect caused by the interactions between neutron events and gamma ray events, and it is intended to be included into a new dosimetry protocol for future 252Cf brachytherapy. PMID- 15353750 TI - Neutron absorbed dose determination by calculations of recoil energy. AB - The aim of this work is to calculate the absorbed dose to matter due to neutrons in the 5-150 MeV energy range. Materials involved in the calculations are Al2O3, CaSO4 and CaS, which may be used as dosemeters and have already been studied for their luminescent properties. The absorbed dose is assumed to be mainly due to the energy deposited by the recoils. Elastic reactions are treated with the ECIS code while for the non-elastic ones, a Monte Carlo code has been developed and allowed to follow the nucleus decay and to determine its characteristics (nature and energy). Finally, the calculations show that the absorbed dose is mainly due to non-elastic process and that above 20 MeV this dose decreases slightly with the neutron energy. PMID- 15353751 TI - Narrow beam neutron dosimetry. AB - Organ and effective doses have been estimated for male and female anthropomorphic mathematical models exposed to monoenergetic narrow beams of neutrons with energies from 10(-11) to 1000 MeV. Calculations were performed for anterior posterior, posterior-anterior, left-lateral and right-lateral irradiation geometries. The beam diameter used in the calculations was 7.62 cm and the phantoms were irradiated at a height of 1 m above the ground. This geometry was chosen to simulate an accidental scenario (a worker walking through the beam) at Flight Path 30 Left (FP30L) of the Weapons Neutron Research (WNR) Facility at Los Alamos National Laboratory. The calculations were carried out using the Monte Carlo transport code MCNPX 2.5c. PMID- 15353752 TI - Determination of absorbed dose in the vicinity of a neutron source. AB - A computational method is presented that transfers the spectral neutron fluence from the measuring position to a reference point in a biological sample. The method is applied to the determination of the absorbed dose in adult blood irradiated with monoenergetic neutrons of 0.035 and 14.8 MeV. PMID- 15353753 TI - Self-shielding effects in neutron spectra measurements for neutron capture therapy by means of activation foils. AB - The design and optimisation of a neutron beam for neutron capture therapy (NCT) is accompanied by the neutron spectra measurements at the target position. The method of activation detectors was applied for the neutron spectra measurements. Epithermal neutron energy region imposes the resonance structure of activation cross sections resulting in strong self-shielding effects. The neutron self shielding correction factor was calculated using a simple analytical model of a single absorption event. Such a procedure has been applied to individual cross sections from pointwise ENDF/B-VI library and new corrected activation cross sections were introduced to a spectra unfolding algorithm. The method has been verified experimentally both for isotropic and for parallel neutron beams. Two sets of diluted and non-diluted activation foils covered with cadmium were irradiated in the neutron field. The comparison of activation rates of diluted and non-diluted foils has demonstrated the correctness of the applied self shielding model. PMID- 15353754 TI - The filter/moderator arrangement-optimisation for the boron-neutron capture therapy (BNCT). AB - The paper presents results of the numerical modelling of the fission-converter based epithermal neutron source designed for the boron neutron capture therapy (BNCT) facility to be located at the Polish research nuclear reactor MARIA at Swierk. The unique design of the fission converter has been proposed due to a specific geometrical surrounding of the reactor. The filter/moderator arrangement has been optimised to moderate fission neutrons to epithermal energies and to remove both fast neutrons and photons from the therapeutic beam. The selected filter/moderator set-up ensures both high epithermal neutron flux and suitably low level of beam contamination. Photons originating from the reactor core are almost eliminated what is the exceptional advantage of the proposed design. It yields one order of magnitude lower gamma radiation dose than the maximum allowed dose in such a type of therapeutic facility. The MCNP code has been used for the computations. PMID- 15353755 TI - Dosimetric considerations on TEPC fluka-simulation and measurements. AB - The response of a tissue equivalent proportional counter (TEPC) has been simulated with the Monte Carlo transport code FLUKA. The absorbed dose distribution of lineal energy y has been determined for several monoenergetic photon and neutron sources. The agreement between the calculated results and the measurements carried out with different well-known sources is well demonstrated. Work is in progress in order to evaluate the response of the instrument in the cosmic ray environment. PMID- 15353756 TI - First microdosimetric measurements with a TEPC based on a GEM. AB - A new type of mini multi-element tissue-equivalent proportional counter (TEPC) based on a gas electron multiplier (GEM) has been designed and constructed. This counter is in particular suitable to be constructed with a small sensitive volume so that it can be used for microdosimetry in intense pulsed radiation fields to measure the microdosimetric spectrum in the beam of, for instance, a clinical linear accelerator. The concept lends itself also for a mini multi-element version of the counter to be used for applications in which a high sensitivity is required. In this paper, we present the first microdosimetric measurements of this novel counter exposed to a 14 MeV monoenergetic neutron beam and a californium (252Cf) source for a counter cavity diameter of 1.8 mm simulating 1.0 microm tissue site size. The measured spectra showed an excellent agreement with spectra from the literature. The specific advantages of the TEPC-GEM are discussed. PMID- 15353757 TI - Clusters of ionisation in nanometre targets for propane-experiments with a Jet Counter. AB - Further evidence on the reliability of the device called the Jet Counter (JC) for studying the formation of ionisation clusters at the nanometre level are presented. The new experimental data on the distributions of ionisation cluster size originating from a 2-10 nm size target in propane irradiated by 3.8 MeV alpha particles are described. The JC consists of a pulse-operated valve that injects an expanding jet of propane into an interaction chamber, where a sensitive volume in the form of a cylinder is created. The sensitive volume was irradiated by 3.8 MeV alpha particles. The resulting distribution of ion clusters, ranging from 2 to 10 nm in unit density gas, has been measured. A method of determining the efficiency of registration of single propane ions using an ion detector is described. A method of deconvolution of the measured to true cluster size distributions is also given. Finally, the measured cluster size distributions are compared with modelled distributions based on Monte Carlo calculations. The results for propane together with previous ones for nitrogen indicate the JC to be an efficient tool for the investigation of radiation quality at the nanometre level. PMID- 15353758 TI - Experimental equivalent cluster-size distributions in nanometric volumes of liquid water. AB - Ionisation cluster-size distributions in nanometric volumes of liquid water were determined for alpha particles at 4.6 and 5.4 MeV by measuring cluster-size frequencies in small gaseous volumes of nitrogen or propane at low gas pressure as well as by applying a suitable scaling procedure. This scaling procedure was based on the mean free ionisation lengths of alpha particles in water and in the gases measured. For validation, the measurements of cluster sizes in gaseous volumes and the cluster-size formation in volumes of liquid water of equivalent size were simulated by Monte Carlo methods. The experimental water-equivalent cluster-size distributions in nitrogen and propane are compared with those in liquid water and show that cluster-size formation by alpha particles in nitrogen or propane can directly be related to those in liquid water. PMID- 15353759 TI - Design of a 10 nm electron collector for a track-nanodosimetric counter. AB - Recently we have developed a track-nanodosimetric counter, which is a gas detector that measures the distributions of electrons induced by a charged particle in nanometric volumes of tissue equivalent matter, positioned at different distances from the track. Sites equivalent to 20 and 24 nm were defined by means of an electron collector, which is a system of electrodes enclosing an almost wall-less cylindrical volume. In this paper, we present the design of a new electron collector that is able to simulate a volume as small as 10 nm in diameter. PMID- 15353760 TI - Reconstruction of cluster distributions at 100% detection efficiency for a track nanodosimetric counter through a Bayesian analysis. AB - A Bayesian unfolding has been applied to ionisation distributions due to 5.4 MeV alpha particles in a 20-nm site obtained using Monte Carlo simulations, taking into account different detection efficiency conditions. The ideal case of a target volume with uniform efficiency has been investigated to study the dependence of the reconstruction on prior distributions. A simplified approach has been used to treat the case of a target volume with non-uniformly distributed efficiency, like the sensitive volume of the track-nanodosimetric counter. Our results point out that Bayesian analysis provides a effective tool for reconstructing the true ionisation distributions, well beyond the maximum measured cluster size. PMID- 15353761 TI - Monte-Carlo calculations of radial dose and restricted-let for protons in water. AB - A new Monte-Carlo code for event-by-event simulation of the transport of energetic non-relativistic protons (approximately 0.5-10 MeV) and all their secondary electrons (down to 1 Ry) in both the vapour and liquid phases of water is presented. A unified particle-water inelastic model for both phases of water has been developed based on experimental optical data and elements of the Bethe theory. The model applies to both electrons and heavy-charged particles and is particularly suitable for extension to other media of biological relevance (organic polymers, DNA, etc.). Condensed-phase effects are included in the liquid version (MC4L) by means of the dielectric functions which, essentially, substitute the oscillator-strength used in the vapour version (MC4V). The results in the form of radial dose distributions and spatially restricted linear energy transfer are presented and compared with the literature. PMID- 15353762 TI - Signal transduction and mechanical stress. AB - Bone undergoes a constant process of remodeling in which mass is retained or lost in response to the relative activity of osteoblasts and osteoclasts. Weight bearing exercise-which is critical for retaining skeletal integrity-promotes osteoblast function, whereas a lack of mechanical stimulation, as seen during spaceflight or prolonged bed rest, can lead to osteoporosis. Thus, understanding mechanotransduction at the cellular level is key to understanding basic bone biology and devising new treatments for osteoporosis. Various mechanical stimuli have been studied as in vitro model systems and have been shown to act through numerous signaling pathways to promote osteoblast activity. Here, we examine the various types of stress and the sequential response of transduction pathways that result in changes in gene expression and the ensuing proliferation of osteoblasts. PMID- 15353763 TI - Longevity, quality, and the one-hoss shay. PMID- 15353764 TI - Extrasolar planets. Planet hunting gets rocky as teams clash over small worlds. PMID- 15353765 TI - Developmental biology. Bonemaking protein shapes beaks of Darwin's finches. PMID- 15353766 TI - Virology. Avian flu finds new mammal hosts. PMID- 15353767 TI - Paleontology. 400-million-year-old wounds reveal a time when predators romped. PMID- 15353768 TI - Scientific publishing. Zerhouni plans a nudge toward open access. PMID- 15353769 TI - Meetings. Europe clones U.S. science festival. PMID- 15353770 TI - Presidential appointments. NSF's acting chief facing legal limit on tenure. PMID- 15353771 TI - Academic leaders. Neuroscientist named MIT president. PMID- 15353772 TI - Neurobiology. Making sense of Tourette's. PMID- 15353773 TI - High-energy astrophysics. Telescopes break new ground in quest for cosmic rays. PMID- 15353774 TI - 7th International Congress on Vertebrate Morphology meeting. Newly hatched dinosaur babies hit the ground running. PMID- 15353775 TI - 7th International Congress on Vertebrate Morphology meeting. Tiny salamanders show their teeth. PMID- 15353776 TI - 7th International Congress on Vertebrate Morphology meeting. Snake tartare--quite a bodyful. PMID- 15353777 TI - Disclosure of clinical trials in children. PMID- 15353778 TI - Antidepressants' use in anorexic girls. PMID- 15353779 TI - SSRIs in children and suicide. PMID- 15353780 TI - Disparities in cancer funding. PMID- 15353781 TI - The case against stem cell research. PMID- 15353782 TI - Problems in FBI mtDNA database. PMID- 15353783 TI - Comment on "Inhibition of hepatitis B virus replication by APOBEC3G". PMID- 15353785 TI - Beyond the ivory tower. A world of glass. PMID- 15353786 TI - Molecular biology. Argonaute journeys into the heart of RISC. PMID- 15353787 TI - Physics. Crystalline electron pairs. PMID- 15353788 TI - Medicine. Targeting apoptotic pathways in cancer cells. PMID- 15353789 TI - Ecology. Spite among siblings. PMID- 15353790 TI - Planetary science. Looking into the giant planets. PMID- 15353792 TI - Coming to grips with bone loss. PMID- 15353793 TI - Aging in rhesus monkeys: relevance to human health interventions. AB - Progress in gerontological research has been promoted through the use of numerous animal models, which have helped identify possible mechanisms of aging and age related chronic diseases and evaluate possible interventions with potential relevance to human aging and disease. Further development of nonhuman primate models, particularly rhesus monkeys, could accelerate this progress, because their closer genetic relationship to humans produces a highly similar aging phenotype. Because the relatively long lives of primates increase the administrative and economic demands on research involving them, new emphasis has emerged on increasing the efficient use of these valuable resources through cooperative, interdisciplinary research. PMID- 15353794 TI - What can progeroid syndromes tell us about human aging? AB - Human genetic diseases that resemble accelerated aging provide useful models for gerontologists. They combine known single-gene mutations with deficits in selected tissues that are reminiscent of changes seen during normal aging. Here, we describe recent progress toward linking molecular and cellular changes with the phenotype seen in two of these disorders. One in particular, Werner syndrome, provides evidence to support the hypothesis that the senescence of somatic cells may be a causal agent of normal aging. PMID- 15353795 TI - Evolution of coral pigments recreated. AB - In proteins homologous to the green fluorescent protein (GFP), formation of red fluorescence requires three autocatalytic steps, whereas only two are needed for green fluorescence. Multiple red/green color diversification events in the GFP superfamily may reflect convergent evolution of the more complex three-step pathway. In the great star coral Montastraea cavernosa, a recreated common ancestor of green and red proteins turned out to be green, indicating that in this case red proteins evolved their color independently from most other homologous red proteins. Furthermore, red color appears to have evolved gradually by small incremental transitions. PMID- 15353796 TI - Electrically driven single-cell photonic crystal laser. AB - We report the experimental demonstration of an electrically driven, single-mode, low threshold current (approximately 260 microA) photonic band gap laser operating at room temperature. The electrical current pulse is injected through a sub-micrometer-sized semiconductor wire at the center of the mode with minimal degradation of the quality factor. The actual mode of interest operates in a nondegenerate monopole mode, as evidenced through the comparison of the measurement with the computation based on the actual fabricated structural parameters. As a small step toward a thresholdless laser or a single photon source, this wavelength-size photonic crystal laser may be of interest to photonic crystals, cavity quantum electrodynamics, and quantum information communities. PMID- 15353797 TI - Macroscopic, neat, single-walled carbon nanotube fibers. AB - Well-aligned macroscopic fibers composed solely of single-walled carbon nanotubes (SWNTs) were produced by conventional spinning. Fuming sulfuric acid charges SWNTs and promotes their ordering into an aligned phase of individual mobile SWNTs surrounded by acid anions. This ordered dispersion was extruded via solution spinning into continuous lengths of macroscopic neat SWNT fibers. Such fibers possess interesting structural composition and physical properties. PMID- 15353798 TI - External and internal morphology of the BAR 1002'00 Orrorin tugenensis femur. AB - Late Miocene fossils from the Lukeino Formation in Kenya's Tugen Hills are assigned to Orrorin tugenensis. Of 20 fossils recovered there to date, 3 are proximal femurs. One of these, BAR 1002'00, preserves an intact head connected to the proximal shaft by an elongated neck. Although this fossil is comparable in size to Pan troglodytes, computerized tomography scans of the neck-shaft junction of BAR 1002'00 reveal that the cortex is markedly thinner superiorly than inferiorly, differing from the approximately equal cortical thicknesses observed in extant African apes, approaching the condition in later hominids, and indicating that O. tugenensis was bipedal. PMID- 15353799 TI - Testing predator-driven evolution with Paleozoic crinoid arm regeneration. AB - Regenerating arms of crinoids represent direct evidence of nonlethal attacks by predators and provide an opportunity for exploring the importance of predation through geologic time. Analysis of 11 Paleozoic crinoid Lagerstatten revealed a significant increase in arm regeneration during the Siluro-Devonian. During this interval, referred to as the Middle Paleozoic Marine Revolution, the diversity of shell-crushing predators increased, and antipredatory morphologies among invertebrate prey, such as crinoids, became more common. Crinoid arm regeneration data suggest an increase in nonlethal attacks at this time and represent a causal link between those patterns, which implies an important role for predator-driven evolution. PMID- 15353800 TI - Polymorphism in presolar Al2O3 grains from asymptotic giant branch stars. AB - We report microstructural and isotopic analyses of two presolar Al2O3 grains. Aluminum oxide is important cosmically, because its presence has been detected in the infrared spectra of the circumstellar envelopes of O-rich asymptotic giant branch stars and because it is predicted to be the first solid to condense in these stellar environments. The two grain structures, one corundum and the other amorphous, confirm that asymptotic giant branch stars produce both phases. The variation in structure and Ti content demonstrates that Al2O3 can condense in the absence of TiO2 seed clusters but that Ti may be important in determining the crystal structure. PMID- 15353801 TI - Reverse methanogenesis: testing the hypothesis with environmental genomics. AB - Microbial methane consumption in anoxic sediments significantly impacts the global environment by reducing the flux of greenhouse gases from ocean to atmosphere. Despite its significance, the biological mechanisms controlling anaerobic methane oxidation are not well characterized. One current model suggests that relatives of methane-producing Archaea developed the capacity to reverse methanogenesis and thereby to consume methane to produce cellular carbon and energy. We report here a test of the "reverse-methanogenesis" hypothesis by genomic analyses of methane-oxidizing Archaea from deep-sea sediments. Our results show that nearly all genes typically associated with methane production are present in one specific group of archaeal methanotrophs. These genome-based observations support previous hypotheses and provide an informed foundation for metabolic modeling of anaerobic methane oxidation. PMID- 15353802 TI - Bmp4 and morphological variation of beaks in Darwin's finches. AB - Darwin's finches are a classic example of species diversification by natural selection. Their impressive variation in beak morphology is associated with the exploitation of a variety of ecological niches, but its developmental basis is unknown. We performed a comparative analysis of expression patterns of various growth factors in species comprising the genus Geospiza. We found that expression of Bmp4 in the mesenchyme of the upper beaks strongly correlated with deep and broad beak morphology. When misexpressed in chicken embryos, Bmp4 caused morphological transformations paralleling the beak morphology of the large ground finch G. magnirostris. PMID- 15353803 TI - Molecular shaping of the beak. AB - Beak shape is a classic example of evolutionary diversification. Beak development in chicken and duck was used to examine morphological variations among avian species. There is only one proliferative zone in the frontonasal mass of chickens, but two in ducks. These growth zones are associated with bone morphogenetic protein 4 (BMP4) activity. By "tinkering" with BMP4 in beak prominences, the shapes of the chicken beak can be modulated. PMID- 15353804 TI - Activation of apoptosis in vivo by a hydrocarbon-stapled BH3 helix. AB - BCL-2 family proteins constitute a critical control point for the regulation of apoptosis. Protein interaction between BCL-2 members is a prominent mechanism of control and is mediated through the amphipathic alpha-helical BH3 segment, an essential death domain. We used a chemical strategy, termed hydrocarbon stapling, to generate BH3 peptides with improved pharmacologic properties. The stapled peptides, called "stabilized alpha-helix of BCL-2 domains" (SAHBs), proved to be helical, protease-resistant, and cell-permeable molecules that bound with increased affinity to multidomain BCL-2 member pockets. A SAHB of the BH3 domain from the BID protein specifically activated the apoptotic pathway to kill leukemia cells. In addition, SAHB effectively inhibited the growth of human leukemia xenografts in vivo. Hydrocarbon stapling of native peptides may provide a useful strategy for experimental and therapeutic modulation of protein-protein interactions in many signaling pathways. PMID- 15353805 TI - A small molecule Smac mimic potentiates TRAIL- and TNFalpha-mediated cell death. AB - We describe the synthesis and properties of a small molecule mimic of Smac, a pro apoptotic protein that functions by relieving inhibitor-of-apoptosis protein (IAP)-mediated suppression of caspase activity. The compound binds to X chromosome- encoded IAP (XIAP), cellular IAP 1 (cIAP-1), and cellular IAP 2 (cIAP 2) and synergizes with both tumor necrosis factor alpha (TNFalpha) and TNF related apoptosis-inducing ligand (TRAIL) to potently induce caspase activation and apoptosis in human cancer cells. The molecule has allowed a temporal, unbiased evaluation of the roles that IAP proteins play during signaling from TRAIL and TNF receptors. The compound is also a lead structure for the development of IAP antagonists potentially useful as therapy for cancer and inflammatory diseases. PMID- 15353806 TI - The emergence of competition between model protocells. AB - The transition from independent molecular entities to cellular structures with integrated behaviors was a crucial aspect of the origin of life. We show that simple physical principles can mediate a coordinated interaction between genome and compartment boundary, independent of any genomic functions beyond self replication. RNA, encapsulated in fatty acid vesicles, exerts an osmotic pressure on the vesicle membrane that drives the uptake of additional membrane components, leading to membrane growth at the expense of relaxed vesicles, which shrink. Thus, more efficient RNA replication could cause faster cell growth, leading to the emergence of Darwinian evolution at the cellular level. PMID- 15353807 TI - Iatrogenic subtle acute aortic dissection during coronary angioplasty for in stent restenosis. Value of intravascular ultrasound for diagnosis and management. AB - A patient with a subtle, iatrogenic, type A aortic dissection following a coronary angioplasty is described. Intravascular ultrasound was useful to confirm the diagnosis of the confined aortic dissection and also to guide adequate stent coverage of its coronary entry door. Both transesophageal echocardiography and magnetic resonance imaging failed to detect any significant aortic wall abnormality. PMID- 15353808 TI - Unintended iatrogenic creation of an internal thoracic artery to anterior coronary vein bypass graft with subsequent reoperation and salvage of the internal thoracic arterial conduit. PMID- 15353809 TI - Successful bailout percutaneous coronary intervention for immediate surgical complication. AB - This case report describes a 36-year-old male with acute complete occlusion of his dominant left circumflex coronary artery after mechanical mitral valve replacement. The patient was successfully treated with percutaneous catheter intervention using drug-eluting coronary stents. PMID- 15353810 TI - Coincidence of true and false aneurysm with rupture into the right atrium. PMID- 15353811 TI - Detection and treatment of coronary artery pseudoaneurysms following coronary stent deployment. PMID- 15353812 TI - Spontaneous coronary artery dissection postpartum. PMID- 15353813 TI - Intravascular ultrasound findings in pseudostenosis during coronary angioplasty. PMID- 15353814 TI - Device closure of fenestrated atrial septal aneurysm: difficulties and complications with implantation of two devices. PMID- 15353815 TI - Use of bivalirudin instead of heparin during a percutaneous coronary intervention in a patient with severe thrombocytopenia. PMID- 15353816 TI - Reversal of recurrent thrombocytopenia due to abciximab. PMID- 15353817 TI - Novel use of the ACCUNET embolic protection system during mechanical thombolysis of a prosthetic aortic valve. AB - We present a case of a thrombosed prosthetic aortic valve that failed to respond to high dose intravenous heparin therapy. Thrombolytic therapy was contraindicated because of recent surgery. The valve was effectively treated by mechanical thrombolysis following the deployment of bilateral ACCUNET Embolic Protection Systems. PMID- 15353818 TI - Innovations in compensation modeling. PMID- 15353819 TI - Unstable angina due to stent fracture. PMID- 15353820 TI - The role of percutaneous thrombectomy in the contemporary treatment of acute myocardial infarction. PMID- 15353821 TI - Coronary bifurcation lesions: to stent one branch or both? AB - OBJECTIVE: The purpose of this study was to evaluate two different stent placement techniques for bifurcation lesions: 1) stenting of the main branch and balloon dilatation of the sidebranch versus 2) stenting of both branches. BACKGROUND: Percutaneous coronary intervention (PCI) of coronary bifurcation lesions remains challenging, and limited information is available regarding whether stent placement is necessary in both branches of the bifurcation using bare-metal stents. Methods. We prospectively followed all patients who underwent PCI for symptomatic bifurcation lesions at our center. All patients were carefully followed for subsequent clinical events. RESULTS: Between March 2001 and November 2002, a total of 50 patients were treated with either stenting of both vessels (double stent group; n = 32) or stenting of the parent vessel and balloon angioplasty of the sidebranch (single stent group; n = 18). Optimal angiographic success was 87.5% in the single stent group and 100% in the double stent group (p = 0.1). The post-procedure percent diameter stenosis of the sidebranch vessel was significantly higher in the single stent group (18 +/- 25% versus 4 +/- 8%; p = 0.005). At 6 months, the incidence of clinically driven repeat target lesion revascularization was 37.6% with 2 stents as compared to 5.6% using 1 stent (p = 0.01). Angiographic restenosis was documented in 40.6% using 2 bifurcation stents, as compared to 11% when using 1 stent (p = 0.05). By multivariable analysis adjusted for baseline differences, stenting the sidebranch was a borderline predictor for major adverse cardiac events at 6 months (odds ratio = 10.3; 95% confidence interval, 0.9-116; p = 0.053). CONCLUSION: For the treatment of true bifurcation lesions, a strategy of stenting both vessels using bare metal stents seems to be associated with worse long-term results, as compared to stenting only the parent vessel. PMID- 15353822 TI - Anticoagulation with bivalirudin during percutaneous coronary intervention for ST segment elevation myocardial infarction. AB - OBJECTIVE: The objective of this retrospective analysis of high-risk patients treated with bivalirudin during primary percutaneous coronary intervention (PCI) for ST-segment elevation myocardial infarction (STEMI) was to evaluate the safety and feasibility of direct thrombin inhibitor (DTI) without concomitant glycoprotein (GP) IIb/IIIa inhibition. BACKGROUND: Reperfusion by PCI is the treatment of choice for patients with STEMI. In patients with stable or unstable angina without ST-segment elevation undergoing PCI, bivalirudin was at least as effective as heparin plus GPIIb/IIIa inhibitors in reducing ischemic events and more effective in preventing bleeding. There are no published studies detailing the use of bivalirudin in patients with STEMI. METHODS: From 09/02 to 05/03 at the Heart Care Centers of Illinois, Blue Island, Illinois. Ninety-one consecutive patients with STEMI underwent PCI with or without stent placement. Bivalirudin was administered as a bolus dose (0.75 mg/kg) followed by infusion (1.75 mg/kg/hr) for the duration of the procedure. Outcomes were recorded over a 30-day follow-up period. RESULTS: Patients (n = 91) had several high-risk characteristics (40% female, 30% diabetes mellitus, 21% previous MI and 18% cardiogenic shock). PCI procedures utilized balloons, stents, or a combination of both. Intraaortic balloon pumps were used for 41% and closure devices for 24% of patients. CONCLUSIONS: This evaluation demonstrates excellent TIMI flow without the addition of GPIIb/IIIa inhibitors. The low mortality and complication rates suggest anticoagulation with bivalirudin in patients with STEMI undergoing PCI is feasible and warrants further study in larger controlled trials to evaluate the effectiveness of bivalirudin in this patient population. PMID- 15353823 TI - The cooling effect of coronary blood flow on heart: a new approach. AB - We tested the hypothesis that blood flow through the coronary arteries may have a cooling effect on heart. The difference (?T) of blood temperature between coronary sinus and right atrium was calculated, during and after acute complete occlusion of blood flow in left coronary artery by balloon dilatation and confirmed by a Doppler guidewire. Coronary sinus blood temperature increased during interruption of coronary blood flow with balloon dilatation. PMID- 15353824 TI - Risk of local adverse events following cardiac catheterization by hemostasis device use and gender. AB - Hemostasis devices are primarily used to stop bleeding from the femoral artery catheterization site after cardiac diagnostic or interventional procedures. Studies assessing the effectiveness of hemostatic devices compared to manual compression have produced mixed results regarding the relative risk of serious adverse events, such as hemorrhage. This study assessed the relative risk of serious complications following the use of the two main types of hemostasis devices (as compared with manual compression), and assessed the relative rates of these complications by gender. PMID- 15353825 TI - Safety of vascular closure devices--are women different? PMID- 15353826 TI - Anomalous origin of the right coronary artery from the left coronary sinus is associated with early development of coronary artery disease. AB - BACKGROUND: Anomalous origin of the right coronary artery (RCA) from the left coronary sinus can cause sudden cardiac death. Whether this unique anatomical defect also predisposes to early development of coronary artery disease (CAD) remains uncertain. METHODS: Demography, cardiovascular risk factor profiles and coronary angiograms were reviewed in 1,532 consecutive patients over the past 3 years. RESULTS: Thirteen patients (0.8%) had anomalous RCA from the left coronary sinus, while the rest had normal origin from the right coronary sinus. CAD was found in 9/13 anomalous RCA patients (group A) and in 795/1,519 patients with normal RCA origin (group B). There were no differences in the distributions of age, sex or cardiovascular risk factors between the two groups. The distribution of critical lesions among the 3 major coronary arteries showed no significant differences either. However, among the patients with RCA involvement, the anomalous RCA group was significantly younger (54.8 +/- 4.8 years versus 64.9 +/- 10.1 years; p = 0.022). CONCLUSION: The anomalous RCA from left coronary sinus is associated with early development of CAD. The affected patients are 10 years younger. PMID- 15353827 TI - Percutaneous endovascular occlusion of symptomatic coronary arteriovenous fistulas with cyanoacrylate. AB - We describe four cases with symptomatic coronary artery fistulas that were treated primarily with endovascular cyanoacrylate embolization. Coils were also used as adjunctive embolic agents in two of these cases. All four cases showed symptomatic improvement after closure of the fistulas. Complications occurred in three cases including transient ST-segment elevation in one, symptomatic pulmonary embolization in a second, and transient pleuritic chest pain, pericarditis and acute renal failure in a third. The technical aspects of all four cases are given together with a review of the use of cyanoacrylate as an embolic material. We conclude that cyanoacrylate embolization could be considered as an alternative technique for the endovascular closure of coronary artery fistulas but must also caution that the use of this embolic agent is hazardous and should be restricted to practitioners experienced in its usage. PMID- 15353828 TI - Temporal spectrum of ischemic complications with percutaneous coronary intervention: the ESPRIT experience. AB - We determined the timing of ischemic complications within 30 days after percutaneous coronary intervention (PCI) in patients enrolled in the Enhanced Suppression of the Platelet IIb/IIIa Receptor with Integrilin Therapy (ESPRIT) trial. Complications (death, myocardial infarction [MI], target vessel revascularization) occurred in 178 of 2064 patients (8.6%) within 30 days. More than 85% of complications occurred within the 24 hours following randomization, with the greatest risk hazard at 12-18 hours. Unexpectedly, 31% of patients who ultimately met criteria for an endpoint MI within 24 hours of PCI had completely normal CK-MB concentrations at the first 6-hour measurement. There was no rebound increase in events after cessation of eptifibatide. Treatment benefit persisted to 30 days. Post-procedural MI is often not detected until greater than or equal to 12 hours after PCI. Treatment with a glycoprotein IIb/IIIa inhibitor is the only modifiable parameter that reduces the risk for early ischemic complications. PMID- 15353829 TI - Intracoronary myocardial contrast echocardiography in a patient with drug refractory hypertrophic obstructive cardiomyopathy revealing extensive myocardium at risk for infarction with alcohol septal ablation. PMID- 15353830 TI - Initial experience in the treatment of chronic superficial femoral artery occlusions with fiberoptic guidance technology. AB - BACKGROUND: Although procedural success rates for treating chronic occlusions are relatively high, dissections and perforations often require stenting. METHODS: A fiberoptic guidance system that visualizes lesion characteristics and incorporates a radiofrequency energy source to aid in crossing total occlusions was used in three men and one woman with superficial femoral artery lesions that were 8-50 cm long and 5-144 months old. RESULTS: Five lesions were attempted and successfully crossed, and there were no observed dissections or perforations. Ankle-brachial indexes returned to normal in two of the four patients. The guidance system was useful in visualizing and treating chronic total femoral occlusions. PMID- 15353831 TI - Right atrial ablation in the management of atrial fibrillation: point of view. PMID- 15353832 TI - Coronary artery dissection and perforation complicating percutaneous coronary intervention. AB - Percutaneous coronary intervention (PCI) is widely utilized in the treatment of symptomatic coronary artery disease. Despite its numerous benefits, serious and potentially life-threatening complications of PCI can occur, including iatrogenic coronary artery dissection and perforation. The incidence of these complications has been augmented by the development of coronary interventional devices intended to remove or ablate tissue. We herein review the classification, incidence, pathogenesis, clinical sequelae and management of coronary artery dissection and perforation in the current era. Specifically, the current angiographic classifications of coronary artery dissections and perforations are reviewed. The findings of several recent, large registries of PCI-related coronary artery perforations are summarized. The management of coronary artery dissection and perforation is discussed at length, including the application of newer modalities such as covered stents. PMID- 15353833 TI - The evolving role of glycoprotein (GP) IIb/IIIa receptor blockade during percutaneous coronary intervention of saphenous vein bypass grafts. PMID- 15353834 TI - Asymptomatic acute inferior ST elevation myocardial infarction from thermal injury complicating radiofrequency ablation for atrioventricular re-entrant tachycardia. AB - One of the more common regular supraventricular tachyarrhythmias encountered in clinical practice is atrioventricular nodal re-entrant tachycardia (AVNRT). Catheter-based radiofrequency ablation is becoming a common modality for the treatment of symptomatic tachyarrhythmias. It is safe and effective, but complications can occur. We present a patient with asymptomatic acute inferior ST elevation myocardial infarction from thermal injury following radiofrequency modification of the slow pathway for the treatment of AVNRT. This case illustrates an unusual mechanism leading to acute myocardial infarction as a complication of a commonly performed electrophysiology procedure that should be recognized by both interventionalists and electrophysiologists and treated aggressively with percutaneous revascularization. PMID- 15353835 TI - An image of coronary thrombus mimicking dissection detected by intravascular ultrasound during functional evaluation of an intermediate lesion. AB - We report a case of a patient in whom coronary arterial thrombus was detected by intravascular ultrasound performed because of an angiographically false image of dissection associated with thrombus obtained during functional evaluation for an intermediate lesion and managed with anticoagulation alone. PMID- 15353836 TI - Complete revascularization of total obstruction of both subclavian arteries and descending abdominal aorta by combined surgery and percutaneous transluminal angioplasty. AB - A 48-year-old man was admitted with chest pain, intermittent claudication and right upper extremity weakness. Magnetic resonance angiography revealed total occlusion of both subclavian arteries and descending abdominal aorta below renal artery. End-to-side aorta to bifemoral graft surgery and percutaneous transluminal angioplasty of both subclavian arteries were performed. Follow-up angiography after 6 months revealed patent left subclavian artery but 70% occluded right subclavian artery. Therefore re-intervention was performed at right subclavian artery. PMID- 15353837 TI - Incidence of canine Babesia gibsoni infection and subclinical infection among Tosa dogs in Aomori Prefecture, Japan. AB - To identify the incidence of Babesia gibsoni (B. gibsoni) in Aomori Prefecture, northeastern Japan, dogs with acute B. gibsoni infection were investigated at the Animal Teaching Hospital, Kitasato University, between April 2002 and March 2003. Eighteen dogs with acute B. gibsoni infection were recognized; they were all male dogs of the fighting dog breed Tosa. Their platelet counts were below normal and their packed cell volumes (PCVs) were at various levels. We collected blood samples from 141 Tosa dogs from Aomori Prefecture and used polymerase chain reaction assay to investigate the incidence of subclinical B. gibsoni infection. We also looked into the serological abnormalities associated with thrombocytopenia or anemia in subclinical infection. Forty-one of 87 dogs (47.1%) with histories of dog fighting, and one dog of 54 without a history of dog fighting were positive for B. gibsoni; that is, 42 of 141 dogs (29.8%) showed a positive result. The mean platelet counts of dogs with subclinical infection were significantly lower and levels of anti-platelet IgG were significantly higher than levels for dogs without infection. Anti-erythrocyte membrane IgG levels were significantly higher in dogs with subclinical infections, although mean PCVs were not significantly different. Tosa dogs from Aomori Prefecture, Japan, were highly infected with B. gibsoni subclinically and this pathogen might be successfully transmitted during dog fighting. Dogs with subclinical infections were at risk of chronic thrombocytopenia, which may be due to autoimmune mechanisms. PMID- 15353838 TI - Effect of antimicrobial agents on the production and release of shiga toxin by enterotoxaemic Escherichia coli isolates from pigs. AB - Edema disease (ED) of pigs is an enterotoxaemic disease caused by enterotoxaemic Escherichia coli (ETEEC) infection. Antimicrobial therapy for pigs with ED is controversial because it may induce death of sickish piglets. In this study, we investigated the effects in vitro of 7 antimicrobial agents, ampicillin, gentamicin, colistin, bicozamycin, fosfomycin, sulfamethoxazole-trimethoprim and enrofloxacin, on the release and production of shiga toxin (Stx) 2e by ETEEC strains. We found that more Stx 2e accumulated in the bacterial cells than was released into supernatant. Associated with inhibition of cell wall synthesis, the exposure to ampicillin or fosfomycin increased the release of Stx 2e. The production levels of Stx 2e in all antimicrobial-treated cultures were equal to the level in the control or less than in the control. These results suggest that cell wall synthesis inhibitors, such as ampicillin and fosfomycin, may change for the worse in the signs in ETEEC infectious pigs. On the other hand, gentamicin, colistin, bicozamycin and enrofloxacin may be useful for the treatment of pigs with ED. PMID- 15353839 TI - Changes in peripheral leukocyte subsets in dairy cows with inflammatory diseases after calving. AB - To clarify the cellular immune system in dairy cows with inflammatory diseases after calving, the leukocyte subsets were examined in Holstein dairy cows. Twenty Holstein cows reared in one herd, were used in this study. Nine cows (Group 1) experienced onset of mastitis or puerperal fever within 2 weeks after calving, and the other eleven cows remained healthy (Group 2) after calving. The numbers of CD3(+), CD4(+) and CD8(+) cells tended to be lower in Group 1 than in Group 2 from the day of calving through week 1. These results suggested that the cows with inflammatory diseases might have experienced a decline in T cells by the day of calving, before the onset of disease. PMID- 15353840 TI - An epidemic of caprine arthritis encephalitis in Japan: isolation of the virus. AB - A disease characterized by arthritis of carpal joints and occasionally by pneumonia was seen among goats on a farm in Nagano prefecture of Japan in the summer of 2002. Serological investigation was done on 30 goats, that is one group on the farm by agar gel immunodiffusion tests using maedi-visna virus as the antigen, and 19 goats were positive. The caprine arthritis encephalitis virus genome was detected in peripheral blood leukocytes from several antibody-positive goats. Two goats among them were necropsied and used for pathological tests and virus isolation. The isolated virus was serially propagated in fetal lamb lung cell cultures. Goats inoculated with the culture fluid became antibody positive. These results suggested that a CAEV-infected goat had been introduced accidentally to the farm and that subclinical infection occurred among the flock. PMID- 15353841 TI - U.S.-type Babesia microti isolated from small wild mammals in Eastern Hokkaido, Japan. AB - Our previous report demonstrated that small wild rodents in Japan harbored two types of novel Babesia microti-like parasites (Kobe and Hobetsu types), but not the type widely distributed throughout the temperate zones of North American and Eurasian Continents (U.S. type). In this study, we surveyed small wild mammals collected at various places in the northern part of Japan, seeking for U.S.-type B. microti. A total of 197 small mammals comprising 10 species, Apodemus speciosus, A. argenteus, Clethrionomys rufocanus, C. rutilus, Eothenomys andersoni, Microtus montebelli, Tamias sibiricus, Sorex unguiculatus, S. caecutiens, and Urotrichus talpoides, were examined. Babesia parasites were detected in A. speciosus, C. rufocanus, C. rutilus, M. montebelli, S. unguiculatus, and S. caecutiens by microscopy of blood smears and by PCR targeting babesial nuclear small-subunit rRNA (rDNA) and beta-tubulin genes. Inoculation of their bloods into experimental animals gave rise to 23 parasite isolates, which included 16 from A. speciosus, 4 from C. rufocanus, and 1 each from C. rutilus, M. montebelli and S. unguiculatus. Sequencing analyses of their rDNA and beta-tubulin genes revealed that, of the 23 isolates, 20 and 3 were of Hobetsu and U.S. types, respectively. The U.S.-type B. microti strains isolated in Japan, however, were distinguishable from the isolates in the United States when their beta-tubulin gene sequences and antigen profiles in Western blots were compared. We conclude that U.S.-type B. microti exists in Japan although it has been genetically and antigenically diversified from that distributed in the United States. The results also suggest that not only rodents, but also some insectivores may serve as reservoirs for the agent of human babesiosis. PMID- 15353842 TI - Paragonimus ohirai metacercariae in crabs collected along the Arakawa River in Tokyo, Japan. AB - Brackish water crabs infected with Paragonimus ohirai metacercariae have been reported in various regions in Japan. However, infected crabs have not been identified in Tokyo. We therefore collected the crab, Chiromantes dehaani, between August 2002 and July 2003 from 12 locations along the Arakawa River that flows through Tokyo. Of the 922 captured crabs, 177 (19%) from 6 locations were infected with Paragonimus metacercariae. The prevalence of metacercariae at these 6 locations ranged from 5 to 89%. The number of metacercariae per infected crab ranged from 1 to 190, with an average of 13.1. The morphological features of the metacercariae and of adult worms recovered from test rats infected with metacercariae showed that the metacercariae in the infected crabs were P. ohirai Miyazaki, 1939. The ITS2 sequence data support this conclusion. This paper is the first description of P. ohirai infection of crabs in Tokyo. PMID- 15353843 TI - Development of nitric oxide synthase-immunoreactive nerves in the cerebral arteries of the rat. AB - Development of cerebrovascular nitrergic nerves was investigated in the rat, using immunohistochemistry for nitric oxide synthase (NOS) and quantitative analysis. Cerebral perivascular NOS nerves usually appeared on the walls of both the intracranial part of the internal carotid artery (ICA) and the internal ethmoidal arteries (IEA) at birth. NOS nerves via the IEA grew more rapidly than those via the ICA. They extended over all the major arteries located more rostral than the middle part of the basilar arteries during the third postnatal week, while those from the ICA remained limited to the caudal segment of the anterior circulation and to the rostral segment of the posterior circulation throughout development. The appearance of NOS nerves on the vertebrate artery (VA) was not demonstrated before the third postnatal week, being apparently far late in development as compared to that of the same nerve type on the ICA and IEA. PMID- 15353844 TI - Removal effect of the water purifier for home use against Cryptosporidium parvum oocysts. AB - The removal effects of the faucet mounted type water purifier for home use were examined against Cryptosporidium parvum oocysts. The water purifier is composed of a layer of granular activated carbon and the hollow fiber membrane filter. The cartridges were unused, 25%, 50% and 75% flow down by Arizona-dust of U. S. A. Two respective cartridges were used of the examination. The faucet and the water purifier were connected by anti-pressure tube, and 3.0 x 10(7) oocysts of Cryptosporidium parvum were injected into anti-pressure tube while water was running. Twenty liter of collected purified water was examined under the fluorescent microscope. Any oocysts in the purified water collected from all cartridges were not found. Therefore, we considered this purifier as an effective one in removing Cryptosporidium oocysts from drinking water. PMID- 15353845 TI - Glucose uptake activity in murine red blood cells infected with Babesia microti and Babesia rodhaini. AB - The glucose uptake activity in Babesia rodhaini and B. microti - infected red blood cell (IRBC) was investigated in mice using 2-deoxy-D-glucose (2DOG) and L glucose (L-Glc), a non-metabolizable analogue of D-glucose and non-incorporative glucose to non-infected RBC (NRBC), respectively. The uptake activities of both DOG and L-Glc were higher in IRBCs than those in NRBC. The concentration dependent uptake of 2DOG and L-Glc in both IRBC revealed a linear curve, indicating non-transporter mediated uptake. In addition, B. microti IRBC showed higher 2DOG uptake than B. rodhaini IRBC, whereas no difference was observed in L Glc uptake. These results indicated that some new glucose uptake system, at least two systems, developed in both IRBC. The new systems were sodium independent, non competitive to L-Glc, and sensitive to temperature. One of two systems had no kinetical difference between B. rodhaini and B. microti IRBC, however another one might have higher uptake activity in B. microti IRBC compared to that in B. rodhaini IRBC. PMID- 15353846 TI - Involvement of endogenous vasopressin in high plasma osmolality-induced anorexia via V1 receptor-mediated mechanism. AB - It is known that water deprivation or injection of hypertonic saline induces anorexia. The present study examined the possible involvement of vasopressin in the suppression of food intake during high plasma osmolality. Intraperitoneal injection of vasopressin (20 microg/kg) into male rats significantly suppressed food intake for 1 hr. This anorectic effect of vasopressin was reversed by simultaneous injection of a peptide antagonist for V(1) receptor (40 microg/kg), but not for V(2) receptor (40 microg/kg). Intraperitoneal injection of hypertonic saline (20% NaCl, 2 ml/kg) similarly suppressed food intake for 2 hr, which was associated with a transient increase in plasma vasopressin concentrations. This hypertonic saline-induced suppression of food intake was blocked by a V(1) receptor antagonist. Vasopressin (40 ng/2 microl) directly administered into the third ventricle of the brain also suppressed food intake for 1 hr. These results suggest that vasopressin participates in the suppression of food intake during high plasma osmolality, the action of which is mediated by V(1) receptors in the brain. PMID- 15353847 TI - Molecular characterization of Theileria orientalis piroplasm protein encoded by an open reading frame (To ORF2) in a genomic fragment. AB - In the present study, a novel antigenic protein expressed in the piroplasm stage of Theileria orientalis was characterized. A 4,707 bp genomic fragment amplified by PCR contained two open reading frames (ORFs). The deduced amino acid sequence of the first ORF showed significantly high similarlity to the ubiquitin carboxy terminal hydrolases/proteases while the second ORF (To ORF2) showed homology to several surface antigens of plasmodia. To ORF2 was expressed to determine whether the protein product is expressed by the parasite. In western blot analysis, bovine antiserum from a T. orientalis-infected calf recognized the recombinant protein containing a C-terminal part of the ORF expressed by baculovirus system. Western blot analysis with the anti-To ORF2 mouse serum recognized a 48 kDa protein in T. orientalis piroplasm lysates. Indirect immunofluorescence antibody test by confocal scanning laser microscopic analysis showed that antisera against the recombinant protein recognized T. orientalis piroplasm in the infected erythrocyte. The results from this study indicate that To ORF2 protein is expressed at the piroplasm stage and is immunogenic. This novel antigenic To ORF2 protein could be exploited for vaccine development against bovine piroplasmosis. PMID- 15353848 TI - Sequencing of canine 5-hydroxytriptamine receptor (5-HTR) 1B, 2A, 2C genes and identification of polymorphisms in the 5-HTR1B gene. AB - Polymorphisms of human genes encoding 5-hydroxytriptamine (serotonin) receptors (5-HTRs) are thought to be associated with psychiatric disorders and behavioral traits. In the present study, we searched for corresponding polymorphisms in the dog and compared allelic frequencies for the canine 5-HTR1B, 5-HTR2A, and 5-HTR2C genes among five canine breeds. The canine genes consisted of the following: 5 HTR1B, 1170 bp; 5-HTR2A, 1413 bp; and 5-HTR2C, 1377 bp. All of these genes were highly homologous with the human genes. We found six single nucleotide polymorphisms (SNPs) in the 5-HTR1B gene (G57A, A157C, G246A, C660G, T955C, and G1146C). Genotyping of the respective SNPs revealed that there were inter-breed variations in the genotypes and allelic frequencies for four out of the six identified SNPs, suggesting that further analyses of the polymorphisms of the 5 HTR1B gene would be useful in order to gain an understanding of the genetic background underlying the diversified behavioral traits among canine species. PMID- 15353849 TI - Transvaginal follicle aspiration in Thai swamp buffalo heifers using different vacuum pressures after FSH pretreatment (Bubalus bubalis). AB - The objective of the experiment was to study oocyte recovery by transvaginal, ultrasound-guided, follicle aspiration, from Thai swamp buffalo using different vacuum pressures. Six adult buffalo heifers, aged 2.5-3.0 yrs were treated with a total dose of 280 mg FSH, given twice a day in a divided doses over a three day period (60/60 mg, 50/50 mg, 30/30 mg) at d7 after progesterone implant. Three vacuum pressures were used; 100 (n=12), 80 (n=12) and 60 mmHg (n=12) and all of the pressures were performed in each animal. The animals were treated repeatedly and collection took place using 2 sets of each pressure every 2 months, giving a total of 36 collections from each animal. The oocyte recovery rates from each pressure were 81.2% (69/85) 79.1% (53/67) and 90.3% (93/103) for 100, 80 and 60 mmHg respectively. The number of oocytes collected per donor were 5.33 +/- 3.27, 4.42 +/- 2.71 and 7.75 +/- 4.31 respectively. The quality of the oocytes did not improved with the lower vacuum pressure. In conclusion, the application of FSH pretreatment improves the yield of oocytes from Thai, swamp buffalo heifers after gonadotropin treatment when using the vacuum pressures between 60-100 mmHg. PMID- 15353850 TI - An evaluation of radiographic hepatic size in dogs with portosystemic shunt. AB - Radiographically, the hepatic sizes of portosystemic shunt (PSS) cases were evaluated. In this study the hepatic area was compared in PSS and non-PSS dogs by utilizing the right lateral radiography. The top three breeds of PSS dogs of Maltese, Shih Tzu and Yorkshire Terrier, were included and these dogs had a significantly smaller hepatic area ratio of 46.37 +/- 0.63%, 61.76 +/- 0.78% and 41.59 +/- 0.23% respectively (p<0.05) and the average overall hepatic area in the 3 dog breeds was 47.75 +/- 0.40%. PMID- 15353851 TI - Primary cardiac fibrosarcoma in a dog. AB - A primary cardiac fibrosarcoma in the right atrium of a 6-year-old Chihuahua dog is described. At necropsy, there was a firm, whitish and spherical mass in the right atrium. Histopathologically, the mass had moderate cellularity composed of spindle-shaped cells with scattered multinucleated giant cells. The tumor cells were arranged in interwoven bundles and sheets in the collagenous stroma. No metastases were observed. Ultrastructurally, the tumor cells mainly consisted of fibroblasts. Multinucleated giant cells did not have any certain organelles that would indicate a higher order of differentiation. Primary cardiac sarcomas in dogs are extremely rare. PMID- 15353852 TI - Detection of Cryptosporidium muris type oocysts from beef cattle in a farm and from domestic and wild animals in and around the farm. AB - Cryptosporidium muris type oocysts were detected from 21 of 516 beef cattle in a farm. Then we surveyed Cryptosporidium oocysts in 348 beef and dairy cattle, 500 pigs, 101 dogs, 38 wild animals and 11 zoo-kept animals in and around the farm. Oocysts were detected from only 2 of 25 Japanese field mice, Apodemus speciosus in the same farm. Gene analysis suggested that the oocysts were different from the C. muris type bovine isolate. PMID- 15353853 TI - Fibrinonecrotic rhinitis caused by a concurrent infection of Fusobacterium necrophorum and Arcanobacterium pyogenes in a cow. AB - An 8 year-old cow showing severe dyspnea and nasal mucosal necrosis immediately after parturition was subjected to pathological examination. The principal lesions were fibrinonecrotic rhinitis, necrotic bronchopneumonia and renal infarction. Fusobacterium necrophorum biotype A and Arcanobacterium pyogenes antigens were detected in the nasal and pulmonary lesions. These results suggest that the lesions were caused by a concurrent infection of the detected bacteria and that the pulmonary lesions were caused by the aspiration of infectious materials from the nasal ones. Mucosal coagulative necroses observed as the initial lesions in rhinitis were frequently associated with multiple thrombosis. The findings might suggest that thrombosis played an important role in the development of the nasal lesions. PMID- 15353854 TI - Changes in myocardial beta1-adrenergic receptor and stimulatory G-protein gene expression after chronic treatment with doxorubicin in rat. AB - The gene expression of beta(1)-adrenergic receptor (beta(1)AR) and stimulatory G protein Gsalpha in ventricle after chronic treatment with doxorubicin (DOX) in rat was investigated. The rats were treated with DOX in a dose of 2.5 mg/kg once a week for 5 weeks, the cumulative dose being 12.5 mg/kg. Two weeks after the last injection, the positive inotropic effect of isoproterenol was noticeably decreased in left atrial muscle preparations isolated from DOX-treated rats. Northern blot hybridization showed that the mRNA transcripts of beta(1)AR and Gsalpha, important signal transduction elements for regulating heart rate and contractility, were significantly decreased in the ventricle of DOX-treated rats. Thus, chronic treatment with DOX decreases the gene expression levels of myocardial beta(1)AR and Gsalpha. PMID- 15353855 TI - A case of canine gastrinoma. AB - The dog of this case was a 10-year-old Shih Tzu with refractory vomiting, diarrhea and anorexia. Endoscopy revealed an unclear at gastric angle, a stenosis at pyloric antrum and congestion in duodenal mucosa. Since abnormal shadows of irregular echo-levels were disclosed by pancreas ultrasonography, serum gastrin level was determined with a suspect of gastrinoma. And an increase of serum gastrin was demonstrated. In addition, postmortem histological examination revealed that the pancreatic cells were positive for gastrin. Based on these findings, the dog was diagnosed as pancreatic gastrinoma. PMID- 15353856 TI - Congenital myopathy in Japanese Black calves. AB - Two Japanese Black bull calves from a dam showed muscular weakness and became recumbent after birth. At necropsy, skeletal muscles, including face, neck, body and proximal and distal forelimb and hindlimb were extremely pale in color and edematous. Histopathological examination of skeletal muscles revealed degenerative changes as follows: replacement of muscle with fat, variation in muscle fiber diameter, internal nuclei, central core-like structures, and vacuolar and hyaline degeneration of muscle fibers. Ultrastructurally, the lesions were characterized by focal myofibrillar disorganization with streaming or irregularity of Z bands. The present cases seem to be classified as congenital myopathy based on pathological alterations and age of onset. PMID- 15353857 TI - Laxity of canine hip joint in two positions with computed tomography. AB - Computed tomographic (CT) examination of 20 canine hip joints was carried out in two positions, normal-standing and weight-bearing. In normal (dorsal acetabular rim angle: DARA < 15 degrees ) or slightly abnormal (DARA, 15 degrees to 20 degrees ) hip joints, the values of parameters to laxity were evaluated as more severe in the weight-bearing position. Comparisons of results using various indicators, including the center distance (CD) index, dorsolateral subluxation score, and lateral center edge angle, revealed that the CD index may be a useful marker of functional laxity in the canine hip joint under CT scanning. Further, CT scanning in the weight-bearing position was more sensitive than in the normal standing position for the detection of laxity in hips with normal or only slightly abnormal DARA. PMID- 15353858 TI - Immunohistochemical study of endothelial nitric oxide synthase in the trigeminal ganglia of a crotaline snake Trimeresurus flavoviridis. AB - The immunoreactivity of constitutive endothelial nitric oxide synthase (eNOS) was studied in the trigeminal ganglia (TG) of a crotaline snake, Trimeresurus flavoviridis. eNOS immunoreactivity was found in TG neurons of different sizes. The percentage of eNOS-positive TG neurons was significantly higher in the mandibular division than in the infrared-related divisions, the maxillary division and ophthalmic ganglion (p<0.001). These findings suggest that eNOS in the TG of crotaline snakes is involved in constitutive neurotransmission in the TG, and is minimally involved in processing in the infrared-sensory system. PMID- 15353859 TI - Prevalence of Salmonella spp. in pigs, chickens and ducks in the Mekong Delta, Vietnam. AB - In order to determine the prevalence of Salmonella spp. in domestic animals in 6 provinces of the Mekong Delta, Vietnam, 1,098 fecal or intestinal content samples from pigs, chickens, and ducks were examined in the period from July to October, 2000. Salmonella spp. were isolated from 78 (7.1%) of the total samples, which included 23 (5.2%) of 439 pigs, 24 (7.9%) of 302 chickens, and 31 (8.7%) of 357 ducks. From those samples, 80 Salmonella strains were isolated and 25 serovars were identified. The predominant serovars were S. Javiana, S. Derby, and S. Weltevreden. S. Javiana and S. Weltevreden were detected together in pigs, chickens, and ducks. These results indicate that the serovars of Salmonella are widely distributed in domestic animals in the Mekong Delta, Vietnam. PMID- 15353860 TI - Morphological characteristics of tectal neurons of layer I projecting to the nucleus geniculatus lateralis ventralis in chick. AB - Some visual information is sent to the nucleus geniculatus lateralis ventralis (GLv) via the cells in layer I (I cells) of the tectum in birds and is used for color vision, papillary reflex, and kineoptic functions. To reveal the morphological features of 'I cells' projecting to the GLv, they were retrogradely labeled with DiI (1,1'-dioctadecyl-3, 3, 3', 3'-tetramethylindo-carbocyanine perchlorate) in chicks. Two different types of neurons, 'spear dendritic I cells' and 'forked dendritic I cells' were identified. The former had small spindle-like soma and an apical dendrite extending to the tectal surface, and the latter had somewhat larger triangular or polygonal soma and plural ascending dendrites. Most of the labeled dendritic endings bifurcated horizontally in layer F, and showed the ending patterns similar to the terminals of optic nerve fibers. PMID- 15353861 TI - Molecular survey of Mycoplasma haemofelis and 'Candidatus mycoplasma haemominutum' infection in cats in Yamaguchi and surrounding areas. AB - A molecular survey of hemoplasma (Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum') in Yamaguchi Prefecture and surrounding areas was performed by using molecular methods. PCR-RFLP with HindIII revealed that 2 cats were infected with M. haemofelis, and 16 with 'C. Mycoplasma haemominutum' among 102 randomly selected cats. Partial 16S rRNA gene sequences of M. haemofelis and 'C. Mycoplasma haemominutum' determined in this study showed percent similarities of 98.3-99.8% and 96.4-100%, respectively, with those from other countries. Hemoplasma infections were more frequently detected in free-roaming cats than inside cats. Also, the status of FeLV infection was another significant risk factor for hemoplasma infection. PMID- 15353862 TI - Relationship between intraduodenal 5-hydroxytryptamine release and interdigestive contractions in dogs. AB - BACKGROUND: 5-hydroxytryptamine (5-HT) is released into the intestinal lumen during the fasting state. However, the relationship between the intraduodenal 5 HT and the interdigestive cyclic motor activity in conscious dogs is unclear. AIM: To correlate intraduodenal 5-HT concentrations with the interdigestive gastroduodenal migrating motor complex (MMC). METHODS: 6 dogs were implanted with 2 force transducers for recording gastroduodenal contractions and 2 catheters for measuring duodenal volume by a non-absorbable marker perfusion technique. Intraduodenal 5-HT concentrations were determined by high performance liquid chromatography at 5-min intervals. RESULTS: During fasting, gastroduodenal motor activity cycled as the MMC; luminal 5-HT concentrations and total outputs varied cyclically in temporal association with the MMC. Mean 5-HT concentrations peaked during phase II (P<0.05 vs. phase I and III), and 5-HT outputs during phases II or III were greater than during phase I (P<0.05). Exogenous motilin (0.3 microg/kg-hr, IV) stimulated 5-HT release into the duodenal lumen with peak values (P<0.05) during motilin-induced phase II and III. Gastroduodenal motor activity was not altered, however, during exogenous intraduodenal administration of 5-HT (300 ng/mL-min). CONCLUSIONS: 5-HT is released cyclically into the duodenal lumen in close temporal association with the MMC, but its physiologic significance in regulation of gastroduodenal motility is unknown. PMID- 15353863 TI - Effects of metabolic inhibition on phosphorylation levels of PKC isoforms in the guinea pig taenia caeci. AB - We investigated which isoform of protein kinase C (PKC) is responsible to metabolic inhibition in the guinea pig taenia caeci with respect to their phosphorylation levels. By Western blot analysis using isoform-specific antibodies, at least four isoforms of PKC, alpha, beta2, epsilon and zeta were identified in the taenia. Prolonged metabolic inhibition of hypoxia, hypoxia+glucose depletion, and addition of cyanide (all in the presence of high K+) for more than 60 min, but not glucose-depletion only, elicited dephosphorylation of PKCs, alpha, beta2 and epsilon, except zeta. Ca2+ depletion from the medium prevented the dephosphorylation of PKCs induced by hypoxia, and apparently inhibited the dephosphorylation induced by hypoxia+glucose depletion. Acute treatment with hypoxia for 10-30 min elicited a gradual dephosphorylation of PKCbeta2, but not of other tested PKC isoforms. Considering the ATP level under various metabolic conditions reported previously, PKCbeta2 is suggested to be primarily responsible to hypoxia, and its dephosphorylation is closely associated with the alteration of adenylate compounds in the cell. Re-oxygenation after prolonged hypoxia did not restore the phosphoryation level of any tested PKCs, suggesting that the dephosphorylation of PKCs is associated with the irreversible damage of the cell under hypoxia. Presumably, the dephosphorylaton of PKCs, particularly PKCbeta2, plays a role in the signal transduction pathway under metabolic inhibition of the taenia, as reported in proliferative and pathophysiological processes in many other cells. PMID- 15353864 TI - MaxiK channel-triggered negative feedback system is preserved in the urinary bladder smooth muscle from streptozotocin-induced diabetic rats. AB - MaxiK channel, the large-conductance Ca2+-sensitive K+ channel, facilitates a negative feedback mechanism to oppose excitation and contraction in various types of smooth muscles including urinary bladder smooth muscle (UBSM). In this study, we investigated how the contribution of MaxiK channel to the regulation of basal UBSM mechanical activity is altered in streptozotocin-induced diabetic rats. Although the urinary bladder preparations from both control and diabetic rats were almost quiescent in their basal mechanical activities, they generated spontaneous rhythmic contractions in response to a MaxiK channel blocker, iberiotoxin (IbTx). The effect of IbTx on the mechanical activity was significantly greater in diabetic rat than in control animal. Similarly, the basal mechanical activity was increased with apamin, an inhibitor for some types of small conductance Ca2+-sensitive K+ channels, and this effect was more pronounced for diabetic rat. However, in both control and diabetic animals, IbTx action was stronger than that of apamin. Diabetes also enhanced the responses to BayK 8644, an L-type Ca2+ channel agonist. The extent of this enhancement in diabetic bladder vs. control was, however, almost the same as that attained with IbTx. Expression levels for MaxiK channel as well as apamin-sensitive K+ channels and L-type Ca2+ channel were not altered by diabetes, when determined as their corresponding mRNA levels. These results indicate that diabetes can potentially increase the basal UBSM mechanical activity. However, in diabetic UBSM, the main negative-feedback system triggered by MaxiK channel is still preserved enough to counteract the possible enhancement of this smooth muscle mechanical activity. PMID- 15353865 TI - Inhibitory actions of cilostazol on electrical responses of smooth muscle isolated from the guinea-pig stomach antrum. AB - We have investigated the effects of cilostazol, a type III phosphodiesterase inhibitor, on the electrical responses of smooth muscle tissue isolated from the guinea-pig stomach antrum. Cilostazol (10(-5) M) inhibited slow waves recorded from circular muscle cells, but did not significantly alter the pacemaker potentials and follower potentials recorded from myenteric interstitial cells and longitudinal muscle cells respectively. Slow potentials generated in isolated circular muscle bundles without attached myenteric interstitial cells were inhibited by cilostazol (>10(-7) M), while all membrane activities were abolished by 10(-5) M cilostazol. In circular muscle bundles, the input resistance of smooth muscle cells and the refractory period for the generation of slow potentials were not altered during the inhibition of spontaneous activity with cilostazol. While cilostazol at 10(-7) and 10(-6) M did not elevate the tissue content of cyclic AMP, at 10(-5) M cyclic AMP was elevated by about 30%. A similar elevation was also produced by 10(-7) M forskolin. The content of cyclic AMP was not significantly increased in preparations stimulated with 10(-3) M caffeine. The potency for inhibiting slow waves was in the order caffeine (10(-3) M) > forskolin (10(-7) M) > cilostazol (10(-5) M). The frequency of slow waves was decreased by caffeine or forskolin but not by cilostazol, while the duration was reduced by caffeine but not by cilostazol or forskolin. Follower potentials were modulated by caffeine and forskolin, but not by cilostazol: the duration was reduced by caffeine, the frequency was reduced by caffeine or forskolin, and the amplitude was not significantly altered by any of them. The results indicate that cilostazol has high selectivity in inhibiting the activity of circular muscle much more than that of longitudinal muscle or pacemaker cells, with no causal relation to the tissue content of cyclic AMP as appears to be the case for the inhibitory actions of caffeine and forskolin. PMID- 15353866 TI - Severity of exercise-induced ischemia with chest pain and recovery from ischemia after the disappearance of chest pain. AB - The severity of exercise-induced painful ischemia and its recovery after the disappearance of pain are unknown. The aim of this study was to investigate the difference in severity of ischemia at both exercise and postexercise between painful ischemia and painless ischemia. After injections of technetium-99m tetrofosmin at peak ergometer exercise and thallium-201 at 3 minutes postexercise, dual-isotope single photon emission tomography was performed in 78 patients with angiographically proven ischemic heart disease. The extent of ischemic areas (the number of areas), the depth of ischemia in the ischemic area (the severity score of ischemia) and the extension of ischemia toward long axis of the left ventricle (the number of left ventricular levels with ischemic areas in apical, middle, and basal levels) at both exercise and postexercise were compared on the basis of the presence of pain and a history of diabetes mellitus (DM). The symptoms improved within 3 minutes postexercise in all painful ischemia patients. Of 59 patients with reversible ischemia, except for 4 painful ischemia patients with DM, the extent and depth of ischemia at postexercise were more severe in 14 painful ischemia patients without DM and 13 painless ischemia patients with DM than 28 painless ischemia patients without DM (extent; 2.9 +/- 1.7 areas, 3.5 +/- 2.8 areas versus 1.4 +/- 1.8 areas, P = 0.005, depth; 3.8 +/- 3.1 scores, 5.8 +/- 5.4 scores versus 1.9 +/- 3.0 scores, P = 0.0084, respectively) despite a comparable severity of ischemia at peak exercise (extent; 5.4 +/- 2.6 areas, 6.0 +/- 2.4 areas versus 4.3 +/- 3.3 areas, depth; 9.3 +/- 5.7 scores, 10.7 +/- 7.3 scores and 7.5 +/- 8.1 scores, all NS). The extension of ischemia toward long-axis of the left ventricle at both peak exercise and postexercise was more severe in the former 2 groups than the latter group (peak exercise; 2.4 +/- 0.6 levels, 2.5 +/- 0.7 levels versus 1.9 +/- 0.8 levels, P = 0.0263, postexercise: 1.8 +/- 0.7 levels, 1.5 +/- 0.9 levels versus 0.8 +/- 0.8 levels, P = 0.0014, respectively). The presence of chest pain is related to the extension of ischemia toward long-axis of the left ventricle, and the disappearance of pain was not related to the recovery of ischemia. PMID- 15353867 TI - Procedural results and distal embolization after saphenous vein graft stenting and angioplasty for in-stent restenosis of grafts. AB - Saphenous vein graft (SVG) angioplasty is associated with frequent periprocedural complications due to distal embolization and a high risk of restenosis. The purpose of this single-center, retrospective study was to determine the distal embolization incidences and outcomes of stenting for SVG lesions and percutaneous angioplasty for in-stent restenosis of these SVGs. We studied 48 consecutive patients (mean age, 62 +/- 7 years, 92% men) who had prior CABG and underwent stent deployment to SVG lesions detected at our institution over a period of 4 years. Mean lesion length was 12.4 +/- 3.2 mm. The minimal lumen diameter increased from 0.7 +/- 0.3 mm to 3.2 +/- 0.4 mm after stenting. Distal embolization as no reflow/slow flow phenomenon occurred in 5 (10%) patients. Angiographic success was achieved in 98% of the patients. Procedural success was achieved in 96% of the patients. No reflow/slow flow phenomenon was observed, particularly in patients with acute coronary syndrome. During the follow-up, 11 patients (23%) had angiographic evidence of restenosis. Lesions were treated with balloon angioplasty and the minimal lumen diameter increased from 2.6 +/- 1.1 mm to 3.1 +/- 0.3 mm. The angiographic and procedural success rates were both 100%. There were no cases of "no" reflow/slow flow. Restenosis was particularly frequent in patients with diabetes mellitus, hypercholesterolemia, and acute coronary syndrome. Stent implantation in patients with de novo SVG lesions can be achieved with a high rate of angiographic and procedural success. The distal embolization risk is lower during angioplasty of in-stent restenosis lesions of SVGs compared to de novo SVG lesions. PMID- 15353868 TI - The influence of diabetes mellitus on plaque volume and vessel size in patients undergoing percutaneous coronary intervention. AB - We evaluated the influence of diabetes on plaque volume and vessel size at a reference segment in diabetic patients undergoing percutaneous coronary intervention using both angiograms and quantitative intravascular ultrasound. A total of 344 patients with 449 de novo coronary lesions including 97 diabetics (133 lesions) who underwent elective percutaneous coronary intervention under intravascular ultrasound guidance were included in this study. Eleven diabetic patients (19 lesions) received insulin and 52 patients (77 lesions) oral hypoglycemic drugs. The other 34 patients (37 lesions) received diet/exercise therapy alone. We measured vessel area (VA) and lumen area (LA) at proximal and distal reference segments by intravascular ultrasound, which were averaged. Plaque area (VA-LA) and % plaque area (100 x plaque area/VA) were subsequently calculated. Although VA was similar between diabetic and non-diabetic patients (13.46 +/- 4.49 mm2 in diabetics versus 14.11 +/- 5.24 mm2 in non-diabetics, P = 0.214), LA was smaller (6.51 +/- 2.63 mm2 versus 7.38 +/- 3.08 mm2, P = 0.004) and % PA was larger (50.4 +/- 11.7 versus 46.5 +/- 11.3, P < 0.001) in diabetic patients, especially the group receiving a hypoglycemic drug or insulin. VA, LA, and % PA were similar between patients with and without insulin treatment. These results potentially might cause undersized device selection without intravascular ultrasound guidance. PMID- 15353869 TI - Angiographic restenosis after myocardial bridge stenting. AB - Data on restenosis after stent implantation in myocardial bridges (MB) are very limited. Six-month angiographic results for 12 symptomatic patients who underwent stent implantation for myocardial bridges were compared retrospectively with those of 39 patients who underwent direct stent implantation for de novo atherosclerotic lesions in the left anterior descending artery. Diameter stenosis decreased from 69 +/- 8% to 4 +/- 5% in the MB group and from 79 +/- 8% to 7 +/- 6% in the control group after stent deployment. Systolic narrowing was abolished in all patients with MB. In follow-up, quantitative angiography revealed late loss of 1.8 +/- 1.3 mm in the MB group and 0.9 +/- 0.9 mm in the control group (P = 0.025). The in-stent restenosis rate was also higher in the MB group compared to the control group (67% versus 28%; P = 0.037). Despite favorable immediate results, stent implantation in MBs may not be promising because of the higher in stent restenosis rate compared to stenting in de novo atherosclerotic lesions. PMID- 15353870 TI - Comparison of rate and rhythm control in patients with atrial fibrillation and nonischemic heart failure. AB - Atrial fibrillation (AF) is a very common cardiac arrhythmia with an increased mortality in patients with heart failure. Whether the best therapeutic approach to these patients is to restore sinus rhythm or to adequately control the ventricular rate is still controversial. The aim of this study was to compare both strategies in patients with AF and nonischemic heart failure. One hundred and fifty-four patients with AF duration greater than 48 hours and nonischemic left ventricular dysfunction were randomized either to a rhythm (n = 84) or rate (n = 74) control group. The composite end points of the study were embolism, death, and exercise capacity. The average age of the patients was 61 +/- 10 years in the rhythm control group and 58 +/- 12 years in the rate control group (P = NS). The average follow-up period was 35 +/- 21 months in the rhythm control group and 37 +/- 19 months in the rate control group (P = NS). In the first year of the study, exercise capacity and left ventricular ejection fraction (LVEF) were improved in the rhythm control group compared to the exercise capacity and LVEF of the rate control group (P < 0.0001 and P = 0.0005, respectively). There were no statistically significant differences in the embolic event rate between the two groups (P = NS). The mortality rate, especially for death due to pump failure, was significantly higher in the rate control group at the end of the study (P < 0.0001). Restoring and maintaining sinus rhythm had a beneficial effect on mortality and exercise capacity in patients with nonischemic heart failure and AF. PMID- 15353871 TI - Is carotid atherosclerosis more important in patients with mitral annular calcification than in those without? AB - It has been suggested that mitral annular calcification (MAC) may be a manifestation of generalized atherosclerosis. However, how the incidence and extent of coronary artery disease (CAD) are affected by the coexistence of carotid atherosclerosis (CAS) in patients with versus without MAC have not yet been studied. We studied 101 patients with echocardiographic MAC and 52 controls without MAC to investigate the clinical impact of CAS on the frequency and severity (defined as the number of obstructed vessels) of CAD in patients with MAC. Carotid Doppler ultrasonographic examination was performed on all patients before coronary angiography. In patients with both MAC and CAS, the incidences of CAD and multivessel disease (> or = 2 vessel or left main coronary artery disease) were significantly higher than in the control group with CAS alone (91% versus 68%, P = 0.008 and 76% versus 44%, P = 0.004, respectively). On the other hand, among study and control patients without CAS, although the frequencies of CAD and multivessel disease were higher in patients with MAC, interestingly, the differences were not statistically significant (37% versus 58% and 15% versus 26%, respectively, P > 0.05 for both). Stepwise multiple logistic regression analysis revealed that CAS (P < 0.001), MAC (P < 0.01) and, to a limited extent hypertension (P = 0.054), were independent predictors for the presence of CAD. In conclusion, the coexistence of CAS is more important in patients with MAC than in those without as it provides valuable information about the incidence and severity of underlying CAD. In cases with MAC but without CAS, MAC could be caused by factors other than atherosclerosis. PMID- 15353872 TI - The effects of octreotide in dilated cardiomyopathy: an open-label trial in 12 patients. AB - Octreotide, a somatostatin analogue, has been found effective in the treatment of acromegalic cardiomyopathy. We investigated whether intermittent octreotide therapy had beneficial effects in patients with ischemic or idiopathic dilated cardiomyopathy, which are refractory to conventional therapy. Twelve patients with ischemic or idiopathic dilated cardiomyopathy were enrolled in the study. In addition to conventional treatment, octreotide (first 50 microg and then 25 microg three times per day for 4 days) was administered and repeated after 1, 2, and 3 months. The patients were evaluated 3 times, before and immediately after the first treatment and after 3 months of treatment, using echocardiography, exercise stress testing, ambulatory ECG, right ventricular catheterization, cardiac enzymes, and the Minnesota living with heart failure questionnaire for quality of life. There were no significant changes in parameters after the first treatment. However, after 3 months of treatment, there were significant improvements in the left ventricular ejection fraction, left ventricular posterior wall thickness, hemodynamics, exercise capacity, and quality of life. Additionally, ischemic burden and the number of ventricular premature beats also decreased slightly. Intermittent octreotide therapy led to significant improvements in patients with ischemic and idiopathic dilated cardiomyopathy refractory to conventional treatment. We believe that this therapy should be attempted as an adjunctive therapy in these patients, and that in this respect, randomized, double-blind, clinical, and large-scale studies are required before regular usage is undertaken. PMID- 15353873 TI - A double blind randomized trial to compare the effects of eprosartan and enalapril on blood pressure, platelets, and endothelium function in patients with essential hypertension. AB - The renin-angiotensin system is the major contributor to development of hypertension, atherosclerosis, and many other cardiovascular diseases. Angiotensin II, one of the main effectors of this system, contributes to the pathogenesis of hypertension and plays an important role in monocyte, platelet, and endothelium interactions. The effects on platelet and endothelial function, either by angiotensin converting enzyme inhibitors or angiotensin receptor antagonists, are still not well understood. A double-blind, randomized, prospective trial of either enalapril (10-20 mg daily) or eprosartan (400-800 mg daily) over a 10-week period was conducted in 42 patients (27 males, 15 females). Platelet activation was evaluated by measuring platelet factor 4 (PF-4), beta thromboglobulin (beta-TG), the ratio of platelet factor 4 to beta thromboglobulin, and endothelial function by measuring total plasma nitrate levels, von Willebrand factor (vWF) levels, and blood flow using venous occlusive plethysmography. After a 10-week treatment with enalapril or eprosartan, the sitting blood pressure in both the enalapril group (from 152.2 +/- 18.7 mmHg to 141.9 +/- 23.5 mmHg, P < 0.05) and eprosartan group (from 151 +/- 10.0 mmHg to 142.3 +/- 12.9 mmHg, P < 0.05) was significantly reduced. Significant diastolic blood pressure (DPB) reduction (from 94 +/- 8.7 to 84.5 +/- 9.6 mmHg, P < 0.05) and a greater DBP reduction response were found in the eprosartan group (63% in eprosartan versus 25% in enalapril). Additionally, dose-dependent reductions in the indices of platelet activation and endothelial dysfunction were observed in patients administered high dose treatments of eprosartan and enalapril, and the beneficial effects of these agents were not correlated with the reduction of blood pressure using both agents. Eprosartan is effective and well-tolerated in the treatment of mid-to-moderate hypertension, and the DBP response reduction to eprosartin was better than that to enalapril. A high dose of either eprosartan or enalapril significantly decreased the indices of platelet activation and endothelial dysfunction in hypertensive patients. The benefits of both agents cannot be explained solely by their antihypertensive effects and possibly may be mediated through their unique effect on angiotensin blockade. PMID- 15353874 TI - Effect of renal artery stenting on renal function in patients with ischemic nephropathy. AB - The aim of this study was to evaluate the effects of stenting on blood pressure and renal functions in azotemic patients with proximal/ostial atherosclerotic renal artery stenosis. Thirteen azotemic patients (5 females, 8 males, average age, 62.7 +/- 8.3 years) who had renal artery stenosis were included in the study. Their blood pressure, estimated glomerular filtration rate (EGFR), and creatinine levels were measured at baseline and during follow-up. Stents were implanted successfully in all of the cases. The average stent diameter and stent length were 7.2 +/- 0.5 mm and 17.2 +/- 3.4 mm, respectively. Antihypertensive drug was abandoned in 1 (7.6%) patient, reduced in 10 patients (76.9%), and not changed in 2 (15.3%) patients. Significant improvement was observed in the mean serum creatinine level at the 12th month when compared with baseline (2.56 +/- 0.88; 1.83 +/- 0.62, P < 0.001). EGFR was 18.38 +/- 4.64 before the procedure and 22.67 +/- 3.81 during follow-up (P < 0.0001). According to the GFR criteria, renal function was determined to be worse in 1 (7.6%) patient, stabilized in 2 (15.3%), and improved in 10 (76.9%) patients. One patient died during the follow up period. Angiographic restenosis was observed in 2 (15.3%) patients. Follow-up major events were observed in 3 (23%) patients. Stenting azotemic patients with renal artery stenosis is a reliable and effective procedure for achieving an improvement in renal function. PMID- 15353875 TI - Improved systolic ventricular function with normal myocardial mechanics in compensated cardiac hypertrophy. AB - There is still controversy about the relation between changes in myocardial contractile function and global left ventricular (LV) performance during stable concentric hypertrophy. To clarify this, we analyzed LV function in vivo and myocardial mechanics in vitro in rats with pressure overload-induced cardiac hypertrophy. Male Wistar rats (70 g) underwent ascending aortic stenosis for 8 weeks (group AAS, n = 9). LV performance was assessed by transthoracic echocardiography under anesthesia. Myocardial function was studied in isolated papillary muscle preparations during isometric contraction. The data were compared with age- and sex-matched sham-operated rats (group C, n = 9). LV weight to-body weight ratio (C: 2.13 +/- 0.14 mg/g; AAS: 3.24 +/- 0.44 mg/g), LV relative wall thickness (C: 0.18 +/- 0.02; AAS: 0.33 +/- 0.09), and LV fractional shortening (C: 54 +/- 5%; AAS: 70 +/- 8%) were increased in group AAS (P < 0.05). Echocardiographic analysis also indicated a significant association (r = 0.74; P < 0.001) between the percent fractional shortening index and LV relative wall thickness. The performance of AAS isolated muscle revealed that active tension (C: 6.6 +/- 1.7 g/mm2; AAS: 6.5 +/- 1.5 g/mm2) and maximum rate of tension development (C: 69 +/- 21 g/mm2/s; AAS: 69 +/- 18 g/mm2/s) were not significantly different from group C (P > 0.05). In conclusion, compensated pressure-overload myocardial hypertrophy is associated with preserved myocardial function and increased ventricular performance. The improved LV function might be due to the ventricular remodeling characterized by an increased relative wall thickness. PMID- 15353876 TI - Synergistic effect of triptolide and tacrolimus on rat cardiac allotransplantation. AB - Recent studies have shown that triptolide inhibits T cell activation through mechanisms different from those of cyclosporine A and tacrolimus and we postulated that triptolide might have a synergistic effect with tacrolimus to enhance immunosuppression. Using a F344 donor-to-Lewis recipient rat combination, we investigated the immunosuppressive effects of triptolide alone or in combination with tacrolimus on the survival of cardiac allografts. Recipients were treated with placebo, triptolide, tacrolimus, and triptolide in combination with tacrolimus at different doses. The median survival time (MST) was 8 days for placebo; 9.5, 11, 14 and 19 days for triptolide monotherapy at doses of 0.04, 0.08, 0.16, and 0.32 mg/kg/day, respectively, and 11, 13.5, and 52 days for tacrolimus monotherapy at doses of 0.025, 0.05, and 0.1 mg/kg/day, respectively. Tacrolimus 0.025 mg/kg/day combined with triptolide 0.08 and 0.16 mg/kg/day prolonged the MST to 17.5 and 20 days, respectively; while tacrolimus 0.05 mg/kg/day combined with triptolide 0.04, 0.08, and 0.16 mg/kg/day prolonged the MST to 21, 23, and 23 days, respectively. These results suggest that triptolide is a moderately effective immunosuppressive agent. Triptolide combined with a subtherapeutic dose of tacrolimus produced a synergistic effect in prolonging rat cardiac allograft survival. PMID- 15353877 TI - Stenting the undeployed stent. AB - Undeployment of a stent which poses a potential risk for future events may become a serious problem in the catheter laboratory. Herewith, we present a case in which we successfully stented an undeployed stent in the distal right coronary artery. PMID- 15353878 TI - Usefulness of a 6 fr right judkins catheter for mechanically extracting a massive intracoronary thrombus from an ectasic right coronary artery: a report on two different cases of thrombectomy. AB - In order to bail out the slow-flow phenomenon (slow flow) created by a massive thrombus in an ectasic right coronary artery, a thrombus was mechanically extracted with a 6 Fr right Judkins (JR) catheter, which proved to be more useful than a usual thrombectomy using a Rescue PT system catheter (Rescue). In case 1, the Rescue was used in combination with thrombolysis but failed to alleviate the slow flow that was implicated in a large infarction. On the other hand, in case 2, aggressive thrombectomy with a 6 Fr JR catheter with an 8 Fr Amplatz guiding catheter successfully extracted the massive intracoronary thrombus, restoring good coronary flow. Therefore, mechanical extraction with a 6 Fr JR catheter is safe and useful in cases of massive thrombus when diffuse coronary artery ectasia complicates an acute myocardial infarction. In addition, this method should be applicable to cases of acute coronary syndrome with massive thrombus. PMID- 15353879 TI - Anomalous right coronary artery originating from the distal left circumflex artery: single coronary artery with choronic atrial fibrillation. AB - This report describes a patient with a single coronary artery in whom the right coronary artery originated from the distal left circumflex artery. Single coronary artery is a rare congenital anomaly of the coronary circulation which is often associated with other congenital cardiac malformations. This anomaly is thought to be clinically significant especially in patients with atrial fibrillation, although no other associated cardiac anomaly was detected. PMID- 15353880 TI - Fabry disease female proband with clinical manifestations similar to hypertrophic cardiomyopathy. AB - Fabry's disease is an X-linked inborn error of glycosphingolipid catabolism, resulting from a deficiency in alpha-galactosidase A (alpha-Gal A). A 56-year-old Japanese woman was at first suspected of having hypertrophic cardiomyopathy. The patient and her son had alpha-Gal A activity in leukocytes that was remarkably below the limit of controls. DNA analysis of the alpha-Gal A gene revealed a novel missense mutation at codon 19 in exon 1, resulting in leucine-to-proline substitution. As a result she was confirmed as a classic Fabry heterozygote. Recent advances in enzyme replacement therapy can reverse the storage of glycosphingolipids in Fabry's disease. Thus, in patients with cardiac hypertrophy, it is important to differentiate Fabry's disease from other causes of hypertrophy. Therefore, it is necessary to measure alpha-Gal A activity in all suspected cases and to analyze genetic abnormalities in heterozygotes. PMID- 15353881 TI - Experience with nifekalant hydrochloride in a patient with ischemic cardiomyopathy and severe ventricular dysfunction after dor operation. AB - A 52-year-old male with ischemic cardiomyopathy and severe ventricular dysfunction underwent coronary artery bypass grafting and left ventricular reconstruction (Dor operation). The patient developed acute onset of incessant ventricular tachycardia in the early postoperative period that was refractory to therapy with class I antiarrhythmic agents, and multiple attempts at electrical cardioversion were required. A combination of intravenous nifekalant hydrochloride and enteral amiodarone was elected as treatment for this recurrent incessant ventricular tachycardia. Nifekalant hydrochloride was administered as a loading dose (0.3 mg/kg/5 min), followed by an intravenous infusion (0.4 mg/kg/hr). Several days after initiating therapy, the patient no longer experienced episodes of ventricular tachycardia, and there was no compromise in hemodynamics. We conclude that nifekalant hydrochloride is a useful agent for suppression of ventricular tachycardia in patients with severe left ventricular dysfunction, especially during the early postoperative period. PMID- 15353882 TI - A coronary artery fistula with saccular aneurysm mimicking a right atrial cystic mass. AB - A 16 year-old Korean girl with cardiomegaly was found to have a "right atrial cystic mass" on transthoracic echocardiography. An unusual cystic structure made the diagnosis difficult. However, transesophageal echocardiography and multidetector computed tomography revealed a coronary artery fistula with a distal saccular aneurysm involving the interatrial septum and draining into the right atrium. Multidetector computed tomography provided clear anatomic visualization that fully delineated the abnormal structures. Multidetector computed tomography may be considered as a good alternative for transesophageal echocardiography in assessing a tortuous coronary artery fistula and aneurysm with complex anatomy. PMID- 15353883 TI - The case of a cyst hydatid localized within the interatrial septum. AB - The ratio of cardiac involvement of Echinoccocus granulosus is 0.02-2% and although seen rarely, involvement of the interatrial septum has also been reported in the published literature. The present case was a 19-year-old male university student admitted to hospital with complaints of headache and dizziness. Computerized tomography of the cranium revealed a cystic mass located at the frontal region and enucleation of the cyst was performed during surgery. A cystic lesion 5 x 4 cm in size was detected within the interatrial septum on two dimensional transthoracic echocardiography during the postoperative period and the patient was referred to our clinic. Open heart surgery was performed and a hydatid cyst that involved the interatrial septum was enucleated. The cyst wall was sutured to the interatrial septum. No complications developed during the postoperative period. The patient was discharged on the fifth day of hospitalization and medical therapy was started with albendazole. PMID- 15353884 TI - Catheter interventional therapy in an elderly patient with deep vein thrombosis and a brain tumor. AB - A 92-year-old woman with a brain tumor developed swelling of the left lower extremity. Venography showed considerable thrombi from the left common iliac vein to the femoral vein. Following implantation of a temporary inferior vena cava filter, catheter aspiration therapy and catheter-directed thrombolysis were performed. Venography after 3 days showed disappearance of the thrombi and an improvement in vein flow. A permanent inferior vena cava filter was implanted. Local intensive thrombectomy and thrombolysis by catheter together with a temporary inferior vena cava filter were effective treatments in this elderly patient with deep vein thrombosis. PMID- 15353885 TI - A case of massive left atrial lipoma occupying pericardial space. AB - We report a rare case of massive left atrial lipoma occupying pericardial space. A 52-year-old male was admitted because of cardiomegaly of unknown etiology. Computed tomography showed a large epicardial mass located along the anterior surface of the heart, from the diaphragm level through the aortic arch level. The mass showed an attenuation value identical with that of subcutaneous adipose tissue and contained some areas with high density. The mass was not enhanced by contrast media. Histologic examination of the specimen obtained by percutaneous biopsy demonstrated mature adipose tissue. An encapsulated adipose mass weighing 620 g, which originated from the left atrium without any invasion to the pericardium, was completely excised. Microscopic examination revealed mature adipose tissue with partial necrosis, confirming the diagnosis of lipoma. PMID- 15353886 TI - Depressive mixed state: a feature of the natural course of bipolar II (and major depressive) disorder? AB - BACKGROUND: Depressive mixed state (DMX), i.e., a combination of hypomanic and depressive symptoms during the same episode, has recently seen a rebirth of studies after Kraepelin's description. Kraepelin observed, in an inpatient sample, that DMX was related to the number of episodes and to duration of manic depressive insanity (illness). STUDY AIM: The aim was to test Kraepelin's observations in a very different sample. METHODS: 563 consecutive outpatients with major depressive episode (MDE)--320 with bipolar II disorder and 243 with major depressive disorder--were interviewed, in a private practice, by the Structured Clinical Interview for DSM-IV as modified by Benazzi and Akiskal (2003). Hypomanic symptoms during MDE were systematically assessed in patients while off psychoactive drugs. DMX was dimensionally defined, following Akiskal and Benazzi (2003), as an MDE plus 3 or more combined hypomanic symptoms. Kraepelin's examples of DMX, i.e., agitated depression (MDE plus psychomotor agitation) and depression with flight of ideas (MDE plus racing thoughts) were also tested. Univariate and multivariate logistic regression was used to study associations. RESULTS: DMX was present in 49.5% of the patients. Multiple logistic regression of DMX versus MDE recurrences and duration of illness, controlled for age, found a strong and significant association only between DMX and duration of illness. The same association was found between agitated depression and duration of illness (but not in depression with racing thoughts). LIMITATION: There was only a single interviewer. DISCUSSION: The findings support Kraepelin's observation of a link between DMX and durations of illness, but not that between DMX and recurrences. Onset of DMX could be more related to the natural course of manic-depressive illness than to a kindling process. PMID- 15353887 TI - Psychopathy in violent female offenders in Finland. AB - BACKGROUND: The base rate and construct of psychopathy in violent female offenders were examined. SAMPLING AND METHODS: More than half of all violent female offenders in Finland who where either hospitalised (n=12) or incarcerated (n=46) during the year of the study were assessed by the Hare Psychopathy Checklist-Revised (PCL-R), Structured Clinical Interview II for DSM-IV, Beck Anxiety Inventory, Beck Depression Inventory-II and Wechsler Adult Intelligence Scale-Revised. RESULTS: PCL-R mean scores and a base rate of psychopathy were found to be in line with earlier studies, showing both lower values and prevalence in females. The PCL-R correlated with symptoms of cluster B personality disorders as well as with depressive personality disorder. Factor 2 was observed to positively correlate with anxiety and depression and negatively with verbal cognitive ability. Logistic regression analysis on the PCL-R items revealed that the items shallow affect, lack of remorse or guilt, conning/manipulation and lack of realistic goals seem to be important indicators of female psychopathy together with callous/lack of empathy, impulsivity and failure to accept responsibility for own actions. CONCLUSION: The finding indicates that the prevalence of psychopathy is lower among female offenders than among male offenders, but further research is needed to examine the base rate of psychopathy in women, and whether the symptoms of psychopathy are expressed differently in women than in men. PMID- 15353888 TI - Bipolar spectrum disorder: a pilot study. AB - OBJECTIVE: To assess depressive features of a proposed definition of bipolar spectrum disorder (BSD). METHODS: Thirty-six patients with bipolar disorder type I or II were compared to 37 patients with unipolar major depressive disorder through patient interview and chart review. RESULTS: Univariate analysis suggests that 7 of 12 (recurrent major depressive episodes, brief major depressive episodes, atypical depressive symptoms, early age of onset, family history of bipolar disorder, antidepressant tolerance, and antidepressant-induced mania) features of major depressive episodes were more likely to occur in bipolar versus unipolar patients. After adjustment in a multivariable regression model, however, the five most powerful predictors of bipolar disorder were brief major depressive episodes, early age of onset, antidepressant- induced mania, postpartum depression, and atypical depressive symptoms. CONCLUSIONS: This preliminary study supports the idea that bipolar disorder is characterized by some depressive features less likely to be found in unipolar depression. Further prospective study needs to be conducted comparing BSD with unipolar depression. PMID- 15353889 TI - Differentiation of cognitive and motor aspects in a digit symbol substitution test in Anorexia nervosa patients, before and after weight restoration. AB - BACKGROUND: In a previous study, using computerized analysis of fine motor behaviour, seriously underweight anorexia nervosa patients in the early phase of treatment were found to have shorter reaction times, but not movement times, in experimental drawing and copying tasks. These differences persisted with weight restoration. SAMPLING AND METHODS: Using computerized analysis of writing and drawing behaviour during the performance of a Digit Symbol Substitution Test, 15 anorexia nervosa patients were compared with 15 normal controls, matched for age, sex and educational level. Patients were retested after weight restoration, as were controls after a similar interval. RESULTS: Patients turned out to be superior in the cognitive, but not in the motor aspects of the Digit Symbol Substitution Test. These differences persisted after weight restoration. A practice effect, due to repeated testing, was found in both groups, which turned out to be cognitive in nature. CONCLUSIONS: The findings show that this new technique can add to the interpretation of classic neuropsychological tests. The results are compatible with previous findings of intact or even superior functioning in anorexia nervosa patients on neuropsychological tasks requiring considerable cognitive effort. PMID- 15353890 TI - Factor analytical study of the short version of the World Health Organization Quality of Life Instrument. AB - The domains of the 26-item World Health Organization Quality of Life Instrument (WHOQOL-Bref) contain heterogeneous items and do not encompass the logical constructs of subjective quality of life (QOL). We compared the WHO 4-domain and 6-domain models of the WHOQOL-Bref with the 8-domain model that we obtained from factor analysis (FA). Data from 118 recently recovered Nigerian psychotic patients were used in confirmatory factor analysis (CFA) to assess goodness of fit and clarity of concept. Our FA model had superior goodness of fit for CFA and provided clarity of concept. Analysis of the WHOQOL-Bref should consider the domains from FA and include 'overall QOL' as an item and dependent variable. Subjective QOL is an aggregate of the following constructs: satisfaction with life circumstances; fulfillment of needs, and opportunity for experience in the milieu. PMID- 15353891 TI - Koro-misidentification syndrome in schizophrenia? A plea for clinical psychopathology. AB - Although current diagnostic procedures favour the diagnosis of similar syndromes in different pathological processes, it cannot be overlooked that many psychiatric symptoms that share a similar 'surface grammar' differ in their 'deep grammar' or structure. In the case presented, we describe an acute psychosis where a Koro-misidentification syndrome might be diagnosed. In the discussion we point out how confusing this approach might be, as well as a feasible way to compensate for the shortcomings of contemporary descriptive psychopathology. PMID- 15353893 TI - A mixture model of arterial growth and remodeling in hypertension: altered muscle tone and tissue turnover. AB - Hypertension results in a thickening of the arterial wall due to a net increase in wall constituents via a coordinated production and removal of smooth muscle and extracellular matrix. Although many reports address the associated changes in material properties, few models address the biomechanics of the growth and remodeling process. In this paper, we employ a new, fundamentally different approach to modeling arterial adaptation in hypertension. In particular, basic, characteristic features of hypertension are simulated using a constrained mixture model wherein individual constituents can turnover at different rates and can have different natural configurations. We show, for example, that if there is no turnover of elastin (which approximates responses in maturity), the model vessel is able to reduce an early elevation in wall stress via a thickening of the wall even though the adaptation may be suboptimal. Conversely, if all constituents can turnover completely (which may approximate late development), the model vessel can restore the state of stress and material properties to native values. Given the potential of such a model, there is a need for more data on the history of turnover of individual constituents and their individual material properties. PMID- 15353892 TI - Pulse pressure-induced transmural fluid flux increases bovine aortic smooth muscle cell apoptosis in a mitogen activated protein kinase dependent manner. AB - BACKGROUND: Mechanical forces associated with blood flow are critical in the regulation of vascular smooth muscle cell (VSMC) growth, migration, differentiation, and apoptosis as fundamental features in the pathogenesis of vascular disease. We investigated the effect of pulse pressure on VSMC apoptosis. METHODS: Using a perfused transcapillary co-culture system, bovine thoracic aortic SMC (BASMC) were exposed to increases in pulsatile flow (0.3-17 ml/min) hence pulse pressure (amplitude of pulse 6-50 mmHg in the absence or presence of bovine aortic endothelial cells (BAEC). The extent of apoptosis was determined by measuring caspase-3 activity, the levels of pro- and anti-apoptotic Bcl-2 family proteins, FasL and cellular apoptosis susceptibility (CAS) protein expression and the extent of DNA fragmentation. RESULTS: Changes in pulse pressure resulted in a significant force- and time-dependent increase in caspase-3 activity in BASMC. This effect was maximal after 6 h, independent of BAEC presence, and attenuated following inhibition of mitogen-activated protein kinase (MAPK) activity with PD98059. In parallel cultures, there was a significant increase in Bad and Bax expression, concomitant with an increase in DNA fragmentation, and a significant decrease in Bcl-2 and Bcl-XL expression. The pro-apoptotic effects of pulse pressure were specific differentiated cells but independent of p53, in as much as FasL and CAS expression were enhanced in differentiated adult but decrease in de differentiated embryonic cells in response to flow. CONCLUSIONS: These results suggest that pulse pressure promotes phenotypically distinct VSMC apoptosis in vitro in an endothelial-independent, MAPK-dependent, manner. PMID- 15353894 TI - Comparative observational studies in therapeutic research: approach to the development of guidelines for research and reporting. Proceedings of a symposium. Ruttihubelbad, Switzerland, 24-27 August 2003. PMID- 15353895 TI - Randomised trials versus observational studies: is the comparative observational study a poor compromise or a perfect synthesis? AB - Overview of some fundamental designs for clinical trials and studies. Some tentative definitions are given in order to stimulate questions and perspectives for further research. PMID- 15353896 TI - Elements of therapeutic research structure. AB - For the purpose of comparative therapeutic research, an overview of some basic terms and definitions is given. The main emphasis lies on the time structure of the research and the possible sources of bias and validity. In addition, methods to deal with confounding, effect modification and possible causal inferences are summarized. PMID- 15353897 TI - A framework for classifying study designs to evaluate health care interventions. AB - Researchers who are trained in epidemiology recognise and use the standard range of designs, i. e. retrospective and prospective cohort studies and case control studies, for studying aetiological questions. The application of study designs to health care interventions is more complex. Researchers may modify familiar designs, in response to specific problems posed by the interventions being evaluated, and an unambiguous nomenclature has not been established. Also, researchers who set out to evaluate interventions are not always trained in epidemiology and often use familiar study design labels in an ambiguous or inappropriate manner. The susceptibility to bias of different study designs is a critical consideration for users of research evidence. Sources of bias are the same for all studies, i. e. selection, performance, detection and attrition bias, but the susceptibility of studies is likely to vary depending on their design. Evidence of associations between features of study designs, specific biases and their consistent influence on effect estimates is lacking. A framework for classifying study design will be proposed, based on key features of study designs, i. e. what researchers actually did. This framework may (a) help to reduce the ambiguity about study design labels, and uncertainty about how a study was actually carried out, and (b) help methodological researchers to gather evidence about associations between different study design features and susceptibility to bias. PMID- 15353898 TI - Randomized and double-blind studies--demands and reality as demonstrated by two examples of mistletoe research. AB - BACKGROUND: Two examples of clinical research with mistletoe extracts were used to demonstrate essential difficulties in carrying out randomized and placebo controlled trials. STUDY 1: In a randomized, placebo-controlled, double-blind study investigating the immunological effects of mistletoe extract, healthy subjects were asked to state whether, in their estimation, they had been treated with verum or a placebo. Due to the intrinsic effects of the mistletoe therapy- local inflammatory reactions at the injection site--100% of the subjects treated with verum and 77% of those treated with a placebo made a correct assessment of their therapy. Although double-blind trials are preferable from the methodological point of view--above all in QoL research--this study shows that double blinding is barely achievable when the investigated therapy has obvious (side) effects. STUDY 2: A prospective, randomized, multicenter study of a mistletoe therapy complementary to chemotherapy treatment of breast cancer had to be stopped after a period of 28 months, because it proved impossible to recruit more than 16 patients in six large study centers. With regard to this example and to other failed, GCP-compliant clinical trials on mistletoe therapy we describe which factors interfere with successful clinical trials. One important point, especially in the investigation of complementary cancer treatments, is that cancer patients are unwilling to have their treatment determined by randomization. Many cancer patients in Germany have their own point of view, as to whether a complementary treatment could be of benefit to them or not. Faced with a life-threatening disease they wish to determine this part of their treatment themselves. CONCLUSION: This background elucidates the need for improving the methodology of non-randomized trials to obtain objective and reliable results even in these fields of clinical research. PMID- 15353899 TI - Retrolective, comparative, epidemiological cohort study with parallel groups design for evaluation of efficacy and safety of drugs with "well-established use". AB - The randomized controlled clinical trial (RCT) is accepted as the "golden standard" for the evaluation of efficacy and safety of new drugs. In contrast, to demonstrate efficacy and safety of drugs with "well-established use" that have been on the European Community market for long time, observational comparative epidemiological studies can be used according to the European drug regulation directive. However, because comparative epidemiological cohort studies can share some risk of bias with other nonrandomized observational study designs, there is a need for an approach that could effectively reduce the bias risk in this type of studies. STUDY OBJECTIVES: The purpose of the study was to evaluate the therapeutic efficacy and safety of a long-term complementary therapy of primary, non-metastatic breast carcinoma patients treated with standardized European mistletoe extract Iscador("mistletoe") in addition to the conventional adjuvant oncologic therapy, and compared to the control group treated with the conventional therapy alone. METHODS: The multicenter, comparative, retrolective, pharmaco-epidemiological cohort study with parallel groups design and randomly selected centers that routinely used both treatments was carried out according to Good Epidemiological Practice rules under a standard operating procedure control. The test group patients received the mistletoe extract treatment subcutaneously for at least 3 months, while the control group patients of the same cohort was exclusively treated with the conventional therapy. The patients were followed up for at least 3 years or until death. The primary endpoint of efficacy was the incidence of adverse reactions to the conventional oncologic therapy. Secondary endpoints were change from baseline of the symptoms associated with the disease and treatment as well as overall survival. All endpoints were adjusted to baseline imbalance and confounders. Safety was assessed descriptively by the number of patients with adverse drug reactions (ADRs) attributed to the test treatment. RESULTS: 1442 patients (710 tests and 732 controls) were eligible for the "per protocol" analysis of efficacy and safety. At baseline, the test group had a more advanced disease and worse prognostic factors profile. After a median follow-up of 66 vs. 60 months, and a median mistletoe therapy duration of 52 months, significantly fewer test group patients (16.2%) than control patients (54.0%) developed ADRs attributed to the conventional therapy [adjusted odds ratio, OR (95% confidence interval, CI), OR = 0.47 (0.32-0.67), p < 0.001]. In the test group, the majority of the symptoms disappeared more frequently, and overall mortality hazard was significantly lower [adjusted hazard ratio, HR (95% CI), HR = 0.46 (0.22-0.96), p = 0.038] than in the control group. Systemic ADRs attributed to the test treatment developed in 0.8%, and local ADRs in 17.3% of the patients. ADR severity was mild to intermediate. Tumor enhancement was not observed. CONCLUSIONS: Complementary therapy of patients with primary, non metastatic breast carcinoma with the mistletoe extract Iscador was safe and in comparison to the control group within the same study cohort showed considerably fewer ADRs attributed to concurrent conventional therapy, reduced disease symptoms, and suggested a significant improvement of survival. Despite some methodical limitations that require careful study planning and conduction as well as critical interpretation, the applied study design seems suitable to evaluate the efficacy and safety of drugs with "well-established use", particularly in oncology. PMID- 15353900 TI - Treatment outcomes in patients with migraine: an ex-post-facto comparison of two in-patient facilities. AB - BACKGROUND: In two hospitals we performed an open, prospective observational study on patients with chronic headache as a measure of internal quality assurance using identical methods. Available data were subordinately analysed in order to compare both studies. QUESTIONS: Are the patient samples of both hospitals comparable? If not, which form of statistical adjustment is recommended? Are there differences in the outcome measures of both facilities? METHODS: The outcomes were defined as differences between baseline values and values at discharge from hospital, respectively 6 months after. Frequency of headache attacks, intensity of pain, intensity of general complaints as well as of concomitant symptoms, and quality of life were determined in advance as dependent variables. To compare both patient groups univariate analysis of variance without and with inclusion of covariates were used. For calculating propensity scores (conditional probability of belonging to one of two groups) a logistic regression with the same covariates serving as independent variables was performed. PATIENTS: 426 patients with the main diagnosis "Migraine" and complete data sets concerning the covariates were selected for analysis. 87% of patients are female, the mean age is 45.5 +/- 11.7 years (range 14-73 yrs). RESULTS: 4 out of 11 potential covariates show statistically significant differences between the patients of both hospitals. Correct classification of patients by means of the propensity score succeeds in 67%. Comparing the outcomes at discharge from hospital, significant differences between both groups exist which are, with one exception, not affected by controlling for covariates. 6 months after discharge two of the initial differences between both patient groups are no longer present. These findings are independent from the statistical technique of risk adjustment. CONCLUSIONS: Because of the observed differences between both patient groups it is recommended to adjust data by regression analysis in order to enhance comparability. The choice for one of the two proposed techniques is secondary. With respect to the analyses clear differences between both hospitals exist in short-term outcomes, disappearing 6 months later. PMID- 15353901 TI - Prospective, comparative cohort studies and their contribution to the benefit assessments of therapeutic options: heart failure treatment with and without Hawthorn special extract WS 1442. AB - BACKGROUND: In addition to testing a drug for its efficacy, pharmacological quality and safety, current policies are increasingly demanding evaluations of the therapeutic benefits provided by a drug in general practice with "non selected" patients and increasingly restrictive economic considerations. OBJECTIVE: One of the trials which addresses this task is the WISO cohort study (Efficacy and socio-economic relevance of treatment of chronic heart failure stage NYHA II with Crataegus extract WS 1442). It compares two different therapeutic strategies in the treatment of heart failure stage NYHA II, i.e. a conventional medication and a therapy which also includes hawthorn special extract WS 1442 (Crataegutt novo 450) in addition to chemical-synthetic drugs. In contrast to clinical trials, the patients in cohort studies are expressly not randomised and the physician in charge independently chooses the administered treatment. This comparative, non-interventional observation provides well-founded evidence of the "real-world effectiveness" of the tested preparation. PATIENTS AND METHODS: 952 patients with heart failure (NYHA II) were enrolled in the study by 217 general practitioners. 588 patients received Crataegus special extract WS 1442 (Crataegutt novo 450) either as an add-on therapy or as a monotherapy (Crataegus cohort) and 364 patients received therapy without hawthorn (comparative cohort). These two groups had the same indication (heart failure NYHA II) but were significantly different regarding gender, age and concomitant cardiovascular disease. Basically, in view of the free choice of therapy made by the physician in charge, such differences are to be expected in comparative observational studies. A sufficient degree of patient comparability was provided by means of the matched-pairs technique, which replaced the randomisation procedure normally used in clinical studies. After 2 years, 130 patient pairs generated by this technique could be included in the interim assessment. RESULTS: The clinical symptoms with regard to all parameters investigated showed the same or a more pronounced improvement in the Crataegus cohort in the course of 2 years. After 2 years, the three cardinal symptoms of heart failure--fatigue (p = 0.036), stress dyspnoea (p = 0.020) and palpitations (p = 0.048)--were significantly less marked in the Crataegus cohort than in the comparative cohort. DISCUSSION: The particular design of the cohort study also provides valuable additional information: (1) Hawthorn special extract WS 1442 was prescribed in registered cardiological practices for the treatment of patients with heart failure stage NYHA II, partly as an alternative and partly as a supplement to the used chemical-synthetic drugs. (2) Favourable effects on the clinical symptoms were achieved although the patients in the Crataegus cohort received markedly fewer chemical-synthetic drugs than the patients in the comparative cohort (ACE inhibitors: 36 vs. 54%, p = 0.004; cardiac glycosides: 18 vs. 37%, p = 0.001; diuretics: 49 vs. 61%, p = 0.061; beta-blockers: 22 vs. 33%, p = 0.052). CONCLUSION: The data show a clear benefit for patients with heart failure stage NYHA II treated with WS 1442. The single or add-on administration in addition to a chemical-synthetic medication resulted in objective improvements at comparable costs. PMID- 15353902 TI - Reasons for caution when evaluating health care interventions using non randomised study designs. AB - The key difference between randomised (RCTs) and non-randomised studies (NRS) is their susceptibility to selection bias. Unlike RCTs, groups in non-randomised cohort studies are unlikely to be balanced because of the reasons leading patients to receive one or another treatment, giving rise to "confounding by clinical indication" (CCI). Researchers can try to minimise the susceptibility of NRS to selection bias both at the design stage, e. g. by matching participants on key prognostic factors, and during data analysis, e. g. by regression modeling. Nevertheless, because of i) logistical difficulties in matching, ii) imperfect knowledge about the relationships between prognostic factors and between prognostic factors and outcome, and iii) because of measurement limitations, it is inevitable that estimates of effect size derived from NRS will be confounded to some extent. Researchers and users of evidence alike need to be aware of the consequences of residual confounding. CCI need not necessarily lead to systematic bias in favour of one treatment but, if CCI acts in an unpredictable way, it will still give rise to additional, non-statistical "uncertainty bias" around the estimate of effect size. PMID- 15353903 TI - Guidelines for reporting non-randomised studies. AB - Non-randomised studies (NRSs) are useful because they allow interventions to be evaluated that are difficult to investigate by randomised controlled trials (RCTs). However, NRSs are more susceptible to bias. The Consolidated Standards of Reporting Trials (CONSORT) statement was established to ensure that researchers report features of RCTs that must be considered when appraising their quality. CONSORT has improved the reporting of key information, highlighting missing key information for users. Researchers have a responsibility to report essential information that allows users to assess the susceptibility of NRS to selection, performance, detection and attrition bias. This paper considers criteria for reporting cohort studies: the rationale behind the CONSORT criteria for reporting of RCTs will be applied to cohort studies. Many of the criteria need no modification but application of others raise difficult issues for cohort studies, e.g.: description and standardisation of control and intervention treatments; description of the method of allocation; choice of prognostic factors to be collected; distinguishing between intended and provided treatments; collection of data on adverse and longterm outcomes; establishing a priori plans for analysis. PMID- 15353904 TI - The importance of considering research design in a wider context. AB - This paper reflects on the importance of considering research design in relation to the question at hand, and argues that the "best" method maximises both rigor and appropriateness. It comments on some of the papers published in this supplement, and emphasises that different audiences will have different needs in terms of the level of evidence required. Suggestions are made how some of the problems associated with randomized trials of Complementary Therapies can be overcome. PMID- 15353905 TI - A new model for olfactory placode development. AB - The peripheral nervous system of vertebrate animals arises primarily from the interaction of cranial neural crest and sensory placodes. Placodes are described as thickenings of ectoderm that arise through cell division during neural tube formation. The olfactory sensory system is one component of the peripheral nervous system that arises from paired sensory placodes during development. The olfactory placodes give rise to the primary sensory neurons, support cells and basal cells of the olfactory epithelium. Recent evidence from work in zebrafish and chick suggests that the olfactory and auditory placodes arise from large fields of cells that converge to form the sensory placode. The olfactory placodes arise from within the neural plate, and cell division is apparent only after the sensory placodes are morphologically distinct. As the olfactory placode is forming, its precursors must segregate from their neighboring fields which will give rise to the adenohypophyseal placode, cranial neural crest, and telencephalon. Analysis has shown that the endocrine cells thought to arise from the olfactory placode originate in the neighboring adenohypophyseal and cranial neural crest domains. The borders separating the domains are plastic, where cells sort as they move, and cell fate is dependent on the identity of neighbors once the cells have converged to form the sensory placode. Thus there is degeneracy built into the system such that cells accommodate changes in the environment until cell migrations controlling the formation of the sensory placodes are complete. PMID- 15353906 TI - Eyes: variety, development and evolution. AB - The selective advantages of using light as a source of information are reflected in the diverse types of extant eyes. The physical properties of light restrict how it can be collected and processed, resulting in only eight known optical systems found in animals. Eyes develop through tissue rearrangement and differentiation. Our understanding of the source of genetic information used in developmental programs is growing rapidly and reveals distributions of gene expression with substantial overlap in both time and space. Specific genes and their products are used repeatedly, making causal relationships more difficult to discern. The phenomenon of groups of genes acting together seems to be the rule. Throughout evolution, particular genes have become associated with distinct aspects of eye development, and these suites of genes have been recruited repeatedly as new eyes evolved. PMID- 15353907 TI - Sensory systems in amphioxus: a window on the ancestral chordate condition. AB - Amphioxus has an assortment of cells and organs for sensing light and mechanical stimuli. Vertebrate counterparts of these structures are not always apparent, and a strong case can be made for homology in only a few instances. For example, amphioxus has anatomically simple but plausible homologs of both the pineal and paired eyes of vertebrates. Placodal and neural crest derivatives are, however, more problematic: the evidence for an olfactory system in amphioxus is only circumstantial and, despite the variety of secondary sensory cell types that occur on the body surface in amphioxus, none are obvious homologs of vertebrate taste buds, neuromasts or acoustic hair cells. A useful perspective can nevertheless be gained by examining differences in amphioxus and vertebrate development, specifically how each specifies and positions sensory precursors, controls their proliferation, and deploys them through the body. The much larger size of vertebrate embryos and the need to cope developmentally with increased scale and cell numbers may account for some key vertebrate innovations, including placodes and neural crest. The presence or absence of specific structural adaptations, like the latter, is therefore less useful for judging homology between amphioxus and vertebrates than shared features of specific cell types. It is also clear that the duration of embryogenesis in vertebrates has been significantly extended in comparison with ancestral chordates so as to incorporate events that would originally have occurred during the post-embryonic growth period, including events of neurogenesis. Consequently, no scenario for the origin of vertebrates can be considered complete unless it deals explicitly with the whole of the life history and changes to it. PMID- 15353908 TI - Lateral line receptors: where do they come from developmentally and where is our research going? AB - The lateral line system is composed of both mechanoreceptors, which exhibit little variation in structure between taxonomic groups, and electroreceptors, which exhibit considerably more variation. Cathodally sensitive ampullary electroreceptors are the primitive condition and are found in agnathans, chondrichthyans, and most osteichthyans. Aquatic amphibians also have ampullary electroreceptors for at least part of their life cycle. The more recently evolved anodally sensitive ampullary electroreceptors and tuberous electroreceptors are only found in four groups of teleost fishes. The basic ontogenetic unit of lateral line development is the dorsolateral placode. Primitively, there are six pairs of placodes, which pass through sequential stages of development into lateral line receptors. There is no question about the origin of primitive mechanoreceptors or electroreceptors, however, we do not have a good understanding of the origin of teleost mechanoreceptors and their ampullary or tuberous electroreceptors; do they come exclusively from dorsolateral placodes or from neural crest or even general ectoderm? A second intriguing lateral line question is how certain teleost fish groups evolved tuberous electroreceptors. Electroreception appears to have re-evolved at least twice in teleosts after being lost during the neopterygian radiation. It has been suggested that the development of tuberous electroreceptors might be due to changes in placodal patterning or a change in the general ectoderm that placodes arise from. Unfortunately, our understanding of lateral line origins in fishes is very sketchy, and, if we are to answer such an evolutionary question, we first need more complete information about lateral line development in a variety of fishes, which can then be combined with gene expression data to better interpret lateral line receptor development. PMID- 15353910 TI - Taste buds: development and evolution. AB - The gustatory system in vertebrates comprises peripheral receptors (taste buds), innervated by three cranial nerves (VII, IX, and X), and a series of central neural centers and pathways. All vertebrates, with the exception of hagfishes, have taste buds. These receptors vary morphologically in different vertebrates but usually consist of at least four types of cells (dark, light, basal, and stem cells). An out-group analysis indicates that taste buds were restricted to the oropharynx, primitively, and that external taste buds, distributed over the head and, in some cases, even the trunk, evolved a number of times independently. The sensory neurons of the cranial nerves that innervate taste buds are believed to arise from epibranchial placodes, which are induced by pharyngeal endoderm, but it has never been demonstrated experimentally that these sensory neurons do, in fact, arise from these placodes. Although many details of the development of the innervation of taste buds are still unknown, it is now clear that taste buds are induced from either ecto- or endodermal epithelia, rather than arising from either placodes or neural crest. At present, there are two developmental models of taste bud induction: The neural induction model claims that peripheral nerve fibers induce taste buds, whereas the early specification model claims that oropharyngeal epithelium is specified by or during gastrulation and that taste buds arise from cell-cell interactions within the specified epithelium. There is now substantial evidence that the early specification model best describes the induction of taste buds. PMID- 15353909 TI - Keeping sensory cells and evolving neurons to connect them to the brain: molecular conservation and novelties in vertebrate ear development. AB - The evolution of the mechanosensory cellular module and the molecular details that regulate its development has included morphological modifications of these cells as well as the formation of larger assemblies of mechanosensory cell aggregates among metazoans. This has resulted in a wide diversity of mechanosensory organs. The wide morphological diversity of organs, including the associated morphological modifications of the mechanosensory cells, suggests parallel evolution of these modules and their associated organs. This morphological diversity is in stark contrast to the molecular conservation of developmental modules across phyla. These molecular data suggest that the evolution of mechanosensory transduction might have preceded that of distinct cellular differentiation. However, once a molecular network governing development of specialized cells involved in mechanosensory transduction evolved, that molecular network was preserved across phyla. Present data suggest that at least the common ancestor of triploblastic organisms, perhaps even the common diploblastic ancestor of bilaterian metazoans, had molecular and cellular specializations for mechanosensation. It is argued that the evolution of multicellular organs dedicated to specific aspects of mechanosensation, such as gravity and sound perception, are evolutionary transformations that build on this conserved molecular network for cellular specialization, but reflect distinct morphological solutions. We propose that the sensory neurons, connecting the craniate ear with the brain, are a derived feature of craniates, and possibly chordates, that came about through diversification of the lineage forming mechanosensory cells during development. This evolutionarily late event suggests a heterochronic shift, so that sensory neurons develop in mammals prior to mechanosensory hair cells. However, sensory neuron development is connected to hair cell development, likely in a clonal relationship. The theme of cellular conservation is reiterated in two examples of chordate otic diversification: the evolution of the horizontal canal system and the evolution of the basilar papilla/cochlea. It is suggested that here again, cellular multiplication and formation of a special epithelium predates the functional transformation to an 'organ' system for horizontal angular acceleration and sound pressure reception, respectively. Overall, evolution of the vertebrate ear needs to be understood as an interplay between and utilization of two gene networks or modules. One is at the level of the molecularly and developmentally conserved mechanosensory cellular module. The other is an increased complexity in the morphology of both adult mechanosensory cells and organs by the addition of end-stage and novel features and associated gene networks to detect specific aspects of mechanosensory stimuli. PMID- 15353911 TI - Effect of irbesartan on the antioxidant defence system and nitric oxide release in diabetic rat kidney. AB - BACKGROUND/AIMS: Increased oxidative stress is involved in the aetiology of diabetic nephropathy, and angiotensin II is reported to play a considerable role in the development of renal damage in diabetic kidney. Angiotensin antagonism can slow the progression of renal impairment in diabetes. The present study was thus designed to examine the effect of an angiotensin II type 1 (AT1) receptor antagonist, irbesartan on renal function, oxidative stress and nitric oxide (NO) release in streptozotocin (STZ)-induced diabetic rats. METHODS: Diabetes was induced by a single intraperitoneal injection of streptozotocin (65 mg/kg) in rats. After 4 weeks of STZ injection, rats were divided into four groups: the control rats, diabetic rats and diabetic rats treated with irbesartan (25 and 50 mg/kg, orally) respectively till 8 weeks starting from 4 weeks after STZ injection. Renal function was assessed by creatinine, blood urea nitrogen, creatinine clearance and urea clearance. Oxidative stress was measured by renal malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and catalase. We also measured renal nitrite levels. RESULTS: At the end of the 8th week, diabetic rats exhibited renal dysfunction as evidenced by reduced creatinine and urea clearance along with enhanced albumin excretion rate as compared with control rats. Biochemical analysis of kidneys revealed a marked increase in oxidative stress demonstrated by increased lipid peroxidation and decreased activities of key antioxidant enzymes, GSH, SOD and catalase in diabetic rats. NO release was also significantly higher in diabetic rats than controls. Chronic treatment with irbesartan in diabetic rats significantly attenuated both renal dysfunction and oxidative stress along with increased NO levels as compared with untreated diabetic rats. The kidneys of diabetic rats showed morphological changes such as hyaline casts, glomerular thickening and moderate interstitial fibrosis and arteriolopathy, whereas irbesartan administration markedly prevented diabetic-induced renal morphological alterations. CONCLUSIONS: The present study suggests that oxidative stress/nitrosative stress is increased in the diabetic kidney and AT1 receptor blockade can prevent these changes. The results also suggest that in STZ-induced diabetic rats, the protective action of irbesartan might be mediated, at least in part, by its effect on tissue oxidant/antioxidant status. PMID- 15353912 TI - Prenatal exposure to high level of glucocorticoids increases the susceptibility of renal proximal tubular cells to apoptosis induced by uropathogenic Escherichia coli toxins. AB - Prenatal exposure to excessive glucocorticoids may alter the developing fetus inducing metabolic and endocrine imbalance in various organs, including the kidney. This study aimed at evaluating whether prenatal exposure to high levels of glucocorticoids adversely affects renal cell survival and predisposes to renal cell death. Pregnant rats were injected with 0.1 mg/kg dexamethasone (DEX) i.p. from day 1 of gestation. Renal proximal tubular cells (PTCs) were prepared from 20-day-old offspring in the DEX (DEX cells) and control groups (CON cells). After 4 days' culture, cells were exposed to uropathogenic Escherichia coli ARD6 toxins at concentrations known to induce apoptotic cell death. We found that cell death rate was significantly higher in DEX than in CON cells. Cells exhibited morphological and biochemical features of apoptosis. Conversely, the activity of the antioxidant enzyme catalase was significantly increased in renal cortex homogenate from 20-day-old DEX rats. The antioxidant vitamin E did not prevent apoptosis. These results indicate that prenatal exposure to high levels of glucocorticoids induces alterations in renal PTCs rendering them more sensitive to E. coli toxins via nonoxidative stress. With the increasing use of multiple doses of glucocorticoids in preterm infants, the possibility that antenatal glucocorticoids may lead to renal adverse consequences is of clinical relevance. PMID- 15353913 TI - Degree of anticoagulation after one subcutaneous and one subsequent intravenous booster dose of enoxaparin: implications for patients with acute coronary syndromes undergoing early percutaneous coronary intervention. AB - BACKGROUND: Many cardiologists continue to be reluctant to utilize low-molecular weight heparin in the treatment of patients with non-ST-segment-elevation acute coronary syndrome because they are concerned about how to manage such patients if they have received only one dose of subcutaneous enoxaparin and are then taken within hours of such treatment to the cardiac catheterization laboratory for percutaneous coronary intervention (PCI). Although we and others have recommended that such patients who have received only one subcutaneous enoxaparin dose receive an intravenous 0.3 mg/kg enoxaparin "booster" dose immediately prior to PCI, there are little actual data to support this recommendation. METHODS: 20 middle-aged subjects were treated with 1 mg/kg subcutaneously-administered enoxaparin and then 6 hours later with a "booster" dose of 0.3 mg/kg intravenously-administered enoxaparin. Anti-Xa levels, as well as ENOX Times, were assessed at baseline, at 2, 4 and 6 hours after the initial SC dose, and at 5 min, 1 and 2 hours after the IV booster dose. RESULTS: At 2 and 6 hours after the initial subcutaneous enoxaparin dose, thirty-five percent of patients had anti-Xa levels below 0.6 IU/mL; twenty percent and ten percent had anti-Xa levels below 0.5 IU/mL at 2 and 6 hours after the initial subcutaneous dose, respectively. After the IV booster dose, all patients had anti-Xa levels in the therapeutic range during the 5 minutes to 2 hours during which blood samples were obtained. There was no significant "overshoot" with this booster dose above what is considered to be the upper therapeutic range. ENOX times showed an overall moderate correlation with anti-Xa levels. CONCLUSIONS: A strategy of administering a 0.3 mg/kg IV booster dose to patients who have received only one subcutaneous dose of enoxaparin and then undergo PCI within the first 2-6 hours of such treatment reliably leads to anti-Xa levels in the therapeutic range. PMID- 15353914 TI - Acute normovolemic hemodilution does not reduce antithrombin concentration for cardiac surgery. AB - BACKGROUND: Acute normovolemic hemodilution (ANH) is used to reduce allogeneic blood transfusion with cardiac surgery. This procedure involves pre-operatively removing and storing a volume of whole blood and replacing the volume with crystalloid. The stored blood is then available for transfusion, if required. Hemodilution associated with ANH may reduce the effectiveness of heparin anticoagulation due to dilution of antithrombin. The aim of this study was to determine if antithrombin concentrations are reduced in patients who undergo one unit of ANH during cardiac surgery. METHODS: Patients scheduled for cardiac surgery (n = 71) were grouped according to whether they did or did not undergo ANH pre-operatively. Antithrombin concentrations were measured before and after ANH. This study had 80% power to detect a difference in reduction of antithrombin concentration of 6% between groups following ANH with an alpha error of <0.05. The effect of one unit ANH was expected to cause a difference of 12% or greater. RESULTS: No significant difference in the concentration of antithrombin between ANH patients and those that did not have ANH, nor was there a difference in the decrease in antithrombin between groups. CONCLUSIONS: The results indicate that one unit of ANH does not significantly reduce the concentration of antithrombin prior to cardiac surgery. Thus patients who undergo one unit of ANH are not at increased risk due to dilution of antithrombin. PMID- 15353915 TI - Association of epicardial and tissue-level reperfusion with left ventricular end diastolic pressures in ST-elevation myocardial infarction. AB - Unfavorable hemodynamics among patients with ST-elevation myocardial infarction (STEMI) have been associated with adverse clinical outcomes and may be linked to a failure to achieve complete reperfusion. We hypothesized that impaired epicardial and tissue-level perfusion after fibrinolytic therapy would be associated with adverse hemodynamics. The relationship between left ventricular end-diastolic pressure (LVEDP), baseline clinical characteristics, and angiographic findings were examined in 666 patients with STEMI treated with fibrinolytic therapy from the TIMI 14, INTEGRITI (TIMI 20), ENTIRE (TIMI 23), and FASTER (TIMI 24) trials. LVEDP was analyzed as a dichotomous variable with an elevated LVEDP defined as LVEDP >18 mmHg (median value). Higher post-fibrinolytic LVEDP was associated with age > or = 65, female gender, Killip Class II-IV on presentation, and LAD culprit location. Elevated LVEDP was associated with both a closed infarct-related artery (58.8% of TIMI Flow Grade (TFG) 0/1 with elevated LVEDP vs. 46.6% of TFG 2/3, p = 0.03) and impaired myocardial perfusion (55.7% of TIMI Myocardial Perfusion Grade (TMPG) 0/1 with elevated LVEDP vs. 43.8% of TMPG 2/3, p = 0.02). In a multivariate analysis, impaired myocardial perfusion (OR 1.7, p = 0.02), abnormal Killip Class (OR 4.8, p = 0.001), age > or = 65 (OR 1.6, p = 0.04), and female gender (OR 1.9, p = 0.01) were independently associated with elevated LVEDP. Elevated LVEDP was independently associated with a greater incidence of in-hospital (OR 11.8, p = 0.02) and 30-day congestive heart failure (OR 4.4, p = 0.02). In STEMI, angiographic indices of incomplete reperfusion are associated with an elevated LVEDP, and elevated LVEDP is associated with adverse clinical outcomes. PMID- 15353916 TI - Haemostatic activity in patients with atrial fibrillation treated with low molecular-weight heparin before and after electrical cardioversion. AB - BACKGROUND: Electrical cardioversion of atrial fibrillation (AF) is associated with a thromboembolic risk, and this risk can be reduced by the use of antithrombotic therapy. International guidelines recommend an effective oral anticoagulant therapy (OAT) for at least 3 weeks before, and 4 weeks after cardioversion. We studied whether electrical cardioversion in it self causes changes in the level of activity in the haemostatic system during treatment with either low-molecular-weight heparin (LMWH). METHODS: Thirty-eight patients with AF were randomised consecutively to either LMWH administered subcutaneous in a fixed daily dose, or conventional OAT. Changes in the biochemical markers prothrombin fragment 1+2 (F1+2), D-Dimer, and soluble fibrin, all reflecting the activity in the haemostatic system, were assessed at baseline, before and after electrical cardioversion in patients treated with LMWH for 3 weeks prior to cardioversion. A follow up compared the time spent on anticoagulation prior to cardioversion, and eventual complications in the two group (LMWH vs. OAT). RESULTS AND CONCLUSIONS: No significant differences between the levels of the biochemical markers measured before, and after cardioversion were seen, indicating that during anticoagulant therapy with LMWH, electrical cardioversion in itself, does not cause an increased activity in the haemostatic system. Also the level of F1+2 had declined significantly after cardioversion, when compared to baseline level in patients, whom had a normal sinus rhythm (NSR) re established. This indicates that even in patients on a stable anticoagulant treatment, restoration of a NSR can cause a further decrease in thrombin generation. The median time spent on antithrombotic treatment prior to cardioversion, was significantly different between the LMWH (27 days) and the OAT group (138 days). Our study indicates that cardioversion in patients on LMWH does not cause a hypercoagulable state and that LMWH significantly shortens the time spent on anticoagulant therapy prior to cardioversion. PMID- 15353917 TI - Myocardial salvage after reduced-dose thrombolysis combined with glycoprotein IIb/IIIa blockade versus thrombolysis alone in patients with acute myocardial infarction. AB - BACKGROUND: The aim of study was to examine the efficacy of reduced-dose alteplase plus abciximab versus alteplase alone by quantifying the amount of myocardium salvaged using myocardial scintigraphy. METHODS: This study analyzed 150 patients with acute myocardial infarction who received alteplase (69 patients) or reduced-dose alteplase plus abciximab (81 patients) in the setting of the Stent versus Thrombolysis for Occluded Coronary Arteries in Patients with Acute Myocardial Infarction (STOPAMI) 1 and 2 trials. Salvage index (proportion of initial perfusion defect salvaged by reperfusion therapy), which was obtained by paired scintigraphic studies performed 7-14 days apart, was the primary endpoint of the study. One-year clinical follow-up was also done. RESULTS: Salvage index did not differ significantly among patients treated with reduced dose alteplase plus abciximab (median, 0.41 [25th; 75th percentiles: 0.13; 0.58]) compared to patients who received alteplase (0.26 [0.09; 0.61], p = 0.30). Final infarct size was 16.0% [4.0; 31.0] of the left ventricle in the group with reduced-dose alteplase plus abciximab and 19.4% [7.9; 34.2] of left ventricle in the group with alteplase (p = 0.44). Within a time-to-admission interval of <2 hours, there was a trend for higher values of salvage index in patients who received reduced-dose alteplase plus abciximab compared with patients who received alteplase (0.55 [0.35; 0.73] versus 0.29 [0.11; 0.69], p = 0.15). For time-to-admission intervals > or = 2 hours, no such trend was observed between those who received reduced-dose alteplase plus abciximab or alteplase (0.25 [0.08; 0.48] versus 0.22 [0.08; 0.46], p = 0.79). Major bleeding occurred in 4 patients (5.0%) in the group with reduced-dose alteplase plus abciximab versus 2 patients (3.0%) in the group with alteplase alone (p = 0.58). CONCLUSION: When used as a general strategy in patients with acute myocardial infarction, adding abciximab to alteplase does not increase significantly the amount of salvaged myocardium as compared with alteplase alone. Combination therapy may offer advantages over thrombolytic agents alone if such therapy is applied within 2 hours from symptom onset; however these data need to be proven by studies of adequate power. PMID- 15353918 TI - Factor V G1691A, prothrombin G20210A, and methylenetetrahydrofolate reductase [MTHFR] C677T gene polymorphism in angiographically documented coronary artery disease. AB - BACKGROUND: Single point mutations in the genes coding for factor V [G1691A; Leiden], prothrombin [PRT; G20210A], and methylenetetrahydrofolate reductase [MTHFR, C677T] were shown to be major inherited predisposing factors for venous thromboembolism. However, their contribution in the development of coronary artery disease [CAD] remains controversial. The aim of the study was to examine the association of these mutations in CAD. METHODS: A total of 96 patients with angiographically-demonstrated CAD [mean age 55.3 +/- 11.3], and 404 healthy subjects [mean age 50.7 +/- 8.9] were recruited into the study. Fasting plasma homocysteine was determined by HPLC, and genotype analysis was assessed by PCR RFLP. RESULTS: The carrier frequency of factor V-Leiden (14.6% vs. 15.1%, p = 0.617) and PRT G20210A (3.1% vs. 3.0%; p = 0.936) were similar between patients and controls, respectively. In contrast, the frequency of the MTHFR variant C677T was 71.9% among patients compared with 45.5% in controls (p < 0.001), of which the T/T genotype was significantly higher among patients (31.3%) than controls (4.5%; p < 0.001). Significantly higher homocysteine levels were seen among T/T genotype in both groups compared to non-T/T carriers (p < 0.05), and among patients compared with controls (18.47 +/- 3.73 micromol/L vs. 16.28 +/- 4.16 micromol/L). In addition, the coexistence of MTHFR C677T with FV-Leiden was seen in 10.4% of CAD patients compared 6.9% of controls (p = 0.001). CONCLUSION: While results from this study clearly demonstrate a strong association of hyperhomocysteinemia and homozygosity of the MTHFR C677T, but not FV-Leiden or PRT G20210A, mutations with confirmed CAD, they also suggest a potential role for factor V-Leiden in MTHFR C677T carriers. PMID- 15353919 TI - Decreased plasminogen activator inhibitor-1 levels in coronary artery aneurysmatic patients. AB - BACKGROUND: Matrix metalloproteinases (MMPs) have been implicated in the pathogenesis of arterial aneurysms through increased proteolysis of extracellular matrix proteins. Increased proteolysis due to elevated matrix degrading enzyme activity in the arterial wall may act as a susceptibility factor for the development of coronary aneurysms. Plasmin strongly stimulates pro-MMP enzyme conversion to the active form. Plasmin hyperactivity due to decreased plasminogen activator inhibitor-1 (PAI-1) may cause MMP over activity and coronary aneurysms. The aim of this study was to investigate the association between PAI-1 and presence of coronary aneurysms. METHODS: Twenty-three patients with aneurysmal coronary artery disease and stable angina were enrolled into study (Group 1). Twenty-two patients without coronary aneurysm were selected as a control group (Group 2). PAI-1 was measured in peripheral venous blood. RESULTS: The plasma PAI 1 level was lower in the coronary artery aneurysmatic patients compared to the control group (8.41 +/- 4.28 vs. 13.32 +/- 10.05 ng/ml, p = 0.037). Serum C reactive protein (CRP) values were not significantly different between groups (3.83 +/- 1.08 vs. 4.01 +/- 1.35 mg/l, p >0.05). CONCLUSION: Increased matrix degrading enzyme activity can cause arterial wall destruction through increased proteolysis of extracellular matrix proteins. Unregulated plasmin hyperactivity due to decreased inhibition by PAI-1 may play an important role in coronary aneurysm formation. PMID- 15353920 TI - Postoperative pulmonary embolism in a young female accompanying with Factor V Leiden mutation and hereditary sypherocytosis. AB - A 20 year-old female, heterozygous for Factor V Leiden mutation (FVLM) is presented. Her personal history was prominent for severe anaemia during her gestation. Aetiology of anaemia was found to be hereditary spherocytosis (HS). Intrauterine foetal death had occurred at 20 weeks of gestational age. Two days after curettage, she developed pulmonary embolism (PE). This is an unusual case of pulmonary embolism and intrauterine foetal death coexisting with FVLM and/or HS. We present the case so that a general practitioner or haematologist can hardly see such cases in daily practice. Hence, a young female with PE should be screened for hypercoagulable states including FVLM or HS. PMID- 15353922 TI - Gambling households in Canada. AB - This paper examines the distribution of gambling dollars in Nova Scotia, Saskatchewan and Canada and studies the impact of this spending on households. We focus first on how gambling expenditures are related to the level and source of household income as well as to other demographic characteristics such as age, education, household composition, geographical area, and sources of income. Next we analyze how gambling expenditures are distributed among those households that gamble. We show how expenditure patterns differ in the intensity of gambling as measured by the proportion of household income or total amount of dollars spent on gambling. Then we study the affects that gambling has on spending on household necessities, changes in net worth, retirement savings and household debt. Finally we determine whether gambling expenditures act as a substitute or a complement to other recreational spending on entertainment products and services. Throughout the paper we offer a comparative analysis of provincial and national data. PMID- 15353921 TI - Resistance to oral vitamin K for reversal of overanticoagulation during Crohn's disease relapse. AB - OBJECTIVE: The purpose of this case report is to describe oral vitamin K resistance in a patient with concomitant Crohn's disease (CD) relapse and supratherapeutic anticoagulation. Additionally, a literature review was conducted to explore the mechanism and supporting evidence for poor response to oral vitamin K during CD relapse. CASE REPORT: A 36 year-old female presented with an elevated International Normalized Ratio (INR) of 7.8 during a relapse of CD including symptoms of severe, persistent diarrhea and reduced appetite. For excessive anticoagulation, initial management consisted of withholding warfarin for seven days, administering vitamin K in a total dose of 10 mg orally and 1 mg intravenously. One week later, the INR remained elevated at 8.09. Subcutaneous vitamin K, in a dose of 5 mg, was administered on day eight, and the INR was reduced to a subtherapeutic result of 1.2 on day eleven. DISCUSSION: The case report illustrates a poor response to recommended and repeated doses of oral vitamin K and a single, small dose of intravenous vitamin K during CD relapse. However, the patient responded favorably to vitamin K by the subcutaneous route. Current literature and consensus guidelines recommend the oral route of vitamin K as first-line management of overanticoagulation due to warfarin. Present data supports that patients with inflammatory bowel disease including CD have a greater incidence of vitamin K deficiency and malabsorption, and this is likely due to multiple pathological mechanisms. CONCLUSIONS: Based on this case report, treatment of overanticoagulation in patients with CD relapse should include aggressive management, close monitoring, and consideration of an alternative, parenteral route of vitamin K administration rather than by the oral route due to potential for poor absorption. PMID- 15353923 TI - Comparing the New Zealand and Swedish national surveys of gambling and problem gambling. AB - National surveys of gambling and problem gambling have recently been completed in New Zealand and Sweden. These studies are unique in that data collection was undertaken by official government statistical agencies, involved large, nationally representative samples, and attained high response rates. Comparison of the findings is facilitated by the use of similar procedures and instrumentation and is of interest in that both countries have similar per capita gambling expenditure and welfare states that have recently undergone major economic and social restructuring. Data on gambling participation, problem gambling prevalence and risk factors for problem gambling are presented and discussed. While there are a number of similarities and differences, the Swedish findings are more similar to those of an earlier national survey conducted in New Zealand during 1991. This suggests that risk factors are changing over time in relation to evolving patterns of gambling participation and attitudes towards gambling, a finding that has implications for future patterns of gambling and problem gambling in these and other countries. PMID- 15353924 TI - The SOGS-RA vs. the MAGS-7: prevalence estimates and classification congruence. AB - The purpose of this study was to compare the prevalence rate estimates and congruence in classification status derived from two popular measures of adolescent gambling (SOGS-RA and MAGS-7). Adolescents from three states (Alabama, Mississippi, and Oregon) completed an anonymous questionnaire ( n =1846 high school students total). Results indicate that the prevalence of probable adolescent pathological gambling varied both as a function of instrument and cut off point utilized for classification (range 1.7%-8.2%). Classification groups (non-problem, at-risk, and problem gamblers) generated by both instruments were found to be associated with reports of gambling frequency, amount of money lost in one gambling occasion, and parental gambling problems. However, concern was raised because the MAGS-7 and the SOGS-RA had little congruence in their three group classification decisions for specific individuals (e.g., only 20.5% agreement for problem gamblers). To improve clinical utility, an empirical case was made for using the SOGS-RA to generate a fourth group of adolescent gamblers, which we labeled "probable pathological gamblers" (SOGS-RA > or = 6). This group was differentiated from the remaining gambling groups on all the validity indices. The implications and limitations of these findings, as well as future directions, are discussed. PMID- 15353926 TI - A science-based framework for responsible gambling: the Reno model. AB - As social observers increasingly identify gambling-related problems as a public health issue, key stakeholders need to join together to reduce both the incidence and prevalence of gambling-related harm in the community. This position paper describes a strategic framework that sets out principles to guide industry operators, health service and other welfare providers, interested community groups, consumers and governments and their related agencies in the adoption and implementation of responsible gambling and harm minimization initiatives. PMID- 15353925 TI - Not the same: a comparison of female and male clients seeking treatment from problem gambling counselling services. AB - Previous studies of problem gamblers portray this group as being almost exclusively male. However, this study demonstrates that females comprised 46% of the population ( n = 1,520) of persons who sought assistance due to concerns about their gambling from the publicly-funded BreakEven counselling services in the state of Victoria, Australia, in one 12-month period. This suggests that the model of service delivery which is community based counselling on a non residential basis may be better able to attract female clients than treatment centres where males predominate such as veterans centres. A comparative analysis of the social and demographic characteristics of female and male gamblers within the study population was undertaken. As with previous studies, we have found significant differences between males and females who have sought help for problems associated with their gambling. Gender differences revealed in this study include females being far more likely to use electronic gaming machines (91.1% vs. 61.4%), older (39.6 years vs. 36.1 years), more likely to be born in Australia (79.4% vs. 74.7%), to be married (42.8% vs. 30.2%), living with family (78.9% vs. 61.5%) and to have dependent children (48.4% vs. 35.7%), than males who present at these services. Female gamblers (A$7,342) reported average gambling debts of less than half of that owed by males (A$19,091). These gender differences have implications for the development and conduct of problem gambling counselling services as it cannot be assumed that models of service which have demonstrated effectiveness with males will be similarly effective with females. PMID- 15353931 TI - Review of evidenced-based practice for the prevention of pressure sores in burn patients. AB - Pressure ulcers represent a complex clinical problem, with a reported incidence of 2.7% to 29.5% in hospitalized patients and an etiology that is multifactorial. The prevention of pressure sores in the burn patient population is clearly an area of practice in need of guidelines for care. A multidisciplinary group of advanced burn care professionals have compiled, critiqued, and summarized herein the current evidence of practice in nursing, nutrition, and rehabilitation as it pertains to the prevention of pressure sores after burn injuries. A broad overview of risk factors and assessment scales is described, and current intervention practices and recommendations for care are provided based, whenever possible, on research findings. In addition, research questions are generated in an attempt to move the specialty of burns toward the formal investigation of pressure sores with the ultimate goal being the development of evidence-based practice guidelines. PMID- 15353932 TI - Temperature threshold for burn injury: an oximeter safety study. AB - Pulse oximeters have become essential devices for evaluating and monitoring patient oxygenation. The probe emits a small amount of heat into the skin in the process of signal detection. Regulations set by the Food and Drug Administration currently limit the maximum allowable temperature of an oximeter probe to 41 degrees C. As a result of the prolonged exposure of extremities to these devices, we sought to determine the actual temperature threshold for burn injury in patients. Eighteen patients undergoing surgery for removal of redundant skin (abdominoplasty, breast reduction) consented to the application of a temperature controlled custom probe with four light-emitting diodes that had temperatures set randomly at the expected threshold for burn injury (42.5 degrees C, 43 degrees C, 43.5 degrees C, and 44 degrees C). The probe was left in place for 8 hours (or less if significant pain was noted). The sites covered by the probes were then checked for signs of injury. On the next day, the redundant skin was removed as a scheduled procedure, and histopathology was performed to detect the extent of burn injury. The study was approved by the local institutional research board. Two patients were excluded because of technical problems with the probe, one of whom had the probe turned off because of pain. The only observed sign of injury was either erythema or a superficial blister that was usually unobservable or slightly red at operation. These subtle signs of a burn were noted in one patient at 43 degrees C, four at 43.5 degrees C, and nine at 44 degrees C. No burns were noted in two patients. Minimal or no signs of injury frequently were noted by histopathology. Pulse oximeter probes are safe up to a temperature of 43 degrees C for at least 8 hours in well-perfused skin. Above that temperature, there is a risk of burn injury. Performing temperature threshold tests in redundant skin that is planned for excision is a potential method for testing the safety of devices or materials. PMID- 15353933 TI - Use of Joshi External Stabilizing System in postburn contractures of the hand and wrist: a 20-year experience. AB - Postburn contractures of the hand and wrist can range from a minor cosmetic problem to a crippling condition. The contractures, initially limited to the skin, extend to all the soft-tissue structures over time, often necessitating capsulotomies and tendon lengthening, although they still may not be amenable to total correction. The Joshi External Stabilizing System (JESS) is a versatile, lightweight external fixator consisting of K wires, distractors, and connecting rods (both hinged and nonhinged) along with various link joints. JESS is a dynamic system that allows the lengthening of the contracted tissues via slow distraction, causing minimal surgical insult. This is a retrospective review of 218 cases of postburn contractures of the hand treated with JESS during the last 20 years. Deformities varied from finger contractures to metacarpophalangeal joint and wrist contractures. All cases were of long duration at the time of presentation with the original injury being 4 months to 20 years old. All were assessed for the degree of function and deformity. The patients' activities of daily living were recorded and x-rays taken to evaluate joint configuration. The patients underwent a conservative surgical release followed by application of the appropriate JESS frame for the correction of the residual deformity. After 6 weeks, the frame was removed and hand therapy continued. An analysis is provided of the outcome. The technique produces functional hands with minimal surgical insult. PMID- 15353934 TI - Treatment of hydrofluoric acid burn to the face by carotid artery infusion of calcium gluconate. AB - Hydrofluoric acid (HF) is highly corrosive substance often used in industrial processes. HF burns to the skin cause local tissue injury. Systemic hypocalcemia may ensue, with the potential to produce life-threatening arrhythmias. Medical treatment consists of local application of topical calcium gels, subcutaneous injection of calcium gluconate, and intravenous or intra-arterial infusion of calcium gluconate. Calcium gluconate infusions have been used for HF burns on distal extremities and digits. We report a case of HF burn to the face that was treated by the use of calcium gluconate infusion via the external carotid artery. PMID- 15353935 TI - Methamphetamine-associated burn injuries: a retrospective analysis. AB - Methamphetamine production and use has increased dramatically during the past 10 years. Methamphetamine production requires combining hazardous and volatile chemicals that expose the manufacturer to burn injuries from explosions and chemical spills. We sought to review the epidemiology of burn injuries in a rural burn center secondary to the use of amphetamine or methamphetamine and/or the manufacture of methamphetamine. Review of the records of 507 patients who were admitted to our burn unit from December 1, 1998, to December 31, 2001, revealed 34 patients who were involved in the use of amphetamines or methamphetamines and/or the manufacture of methamphetamine. Thirty-one patients tested positive for either amphetamine (n = 2) or methamphetamine (n = 29) on routine admission urine drug screens. Twenty of these patients were involved in the manufacture of methamphetamines. Three additional patients were identified as methamphetamine manufacturers but tested negative for the use of methamphetamines. The mean age of the study population was 31.88 +/- 7.65 years, with a male:female ratio of 10.3:1. The average burn size was 18.86 +/- 20.72, with the majority secondary to flame (n = 26). Patient burn admission histories were vague, and the patient's involvement in the manufacture of methamphetamine was often only later confirmed by media, the fire marshal, family members, or the patient. Fifteen patients showed the usual withdrawal pattern of agitation and hypersomnolence, with seven patients requiring detoxification with benzodiazepines. Two were admitted acutely to the psychiatric ward for uncontrollable agitation. Eighteen patients were offered chemical dependency treatment, and two completed therapy. There was one mortality. The mean cost per person was US 77,580 dollars (range, US 112 dollars US 426,386 dollars). The increasing use of and manufacture of methamphetamine presents new challenges for the burn team because these patients can become violent and frequently need assistance with detoxification. Routine drug screens are mandatory in identifying methamphetamine use to alert burn unit personnel to particular management problems and target individuals who may be receptive to drug rehabilitation. PMID- 15353936 TI - Firefighter burn injuries: a 10-year longitudinal study. AB - Our metropolitan area employs approximately 11,000 firefighters who respond to more than 435,000 fire-related incidents per year. It is inevitable that some of these firefighters will suffer burn injuries. This 10-year retrospective review describes the epidemiology of firefighters with burn injuries who were treated at our burn center. From 1992 to 2002, 987 firefighters were treated at our burn center. The total number of firefighters treated for burn injuries and the number of firefighters who were treated for burn injuries to the lower extremities occurred in a bimodal distribution. Injury prevention efforts will continue to further reduce the incidence of burn injuries in the firefighters of our community. PMID- 15353937 TI - A survey of airway and ventilator management strategies in North American pediatric burn units. AB - A survey was used to gather information regarding airway management patterns in thermally injured children. North American pediatric burn centers listed by the American Burn Association were sent a survey designed to examine patterns of pediatric airway management in children with acute respiratory failure. The sample population means for the number of patients ventilated more than 48 hours and the number of patients ventilated more than 48 hours with inhalation injury were used to separate centers into large and small pediatric burn centers. Small pediatric burn centers had less than 50 patients who were intubated during a 5 year period. A five-point nominal scale was used to facilitate statistical analysis. Twenty-five pediatric burn centers included in the analysis estimated that 11,494 children were admitted during the 5-year period. There was no statistically dominant ventilator mode being used in the setting of acute respiratory failure identified by this survey. Large pediatric burn centers reported more frequent use of cuffed endotracheal tubes and more frequent change from an uncuffed to a cuffed endotracheal tube in patients who were difficult to ventilate because of an excess leak. Large pediatric burn centers reported a higher prevalence of tracheomalacia then small pediatric burn centers. Steroids were used by most centers before extubation in patients with persistent airway edema. No centers reported complications from steroid use. There is lack of clear consensus regarding the application of various ventilator modes in the setting of acute respiratory failure irrespective of center volume. There were divergent of practice patterns between large and small pediatric burn centers regarding the use of cuffed endotracheal tubes and the timing of tracheostomy. There was agreement between large and small pediatric burn centers in tracheostomy use in children older the age of 7 and the use of steroids as an adjunct to extubation in patients with lingering airway edema. Pediatric burn patients may benefit from clinical trials that clarify the advantages and disadvantages of various ventilator modes, the use of cuffed tubes, and the timing of tracheostomy. PMID- 15353938 TI - Home treadmill friction injuries: a five-year review. AB - Treadmills are popular home fitness machines in American homes. Young children are at risk for friction injuries if they contact moving treadmills. The purpose of this study was to determine the impact of treatment of treadmill friction injuries in children. A review of 1,211 pediatric patients younger than 6 years treated at the Intermountain Burn Center between July 1997 and June 2002 was conducted. Forty-eight of these cases (4%) were treadmill friction injuries. The mean TBSA of these burns was 0.5%. The volar surface of the hand was the most common site of injury. Twenty-two (46%) of the 48 identified patients had full thickness injuries that were treated surgically. Medical costs associated with treadmill friction injuries averaged US 2,385 dollars. The number of treadmill friction accidents resulting in friction injuries to children less than 6 years of age deserves serious attention and increased public awareness. PMID- 15353939 TI - Effectiveness of a burn prevention campaign for older adults. AB - Older adults are involved in one fifth of burn injury admissions in the Province of Ontario Canada. Most burn injuries in this population occur at home while cooking, bathing, or smoking. The purpose of this study was to evaluate the effectiveness of an educational campaign to improve burn prevention knowledge in older adults of a major metropolitan city. Changes in participants' burn prevention knowledge were determined using standardized precampaign and postcampaign (4-6 weeks) surveys. Of 209 older adult participants, 126 (60.3%) completed the precampaign and postcampaign surveys. There was a significant increase (P <.05) in burn prevention knowledge postintervention. Age, education level, and living conditions did not influence the change in burn prevention knowledge. This burn prevention campaign for older adults was effective in improving burn prevention knowledge, but it remains unclear as to whether this will ultimately result in a change in burn prevention behavior. PMID- 15353940 TI - Car radiator burns: a prevention issue. AB - Scald burns continue to be the major cause of injury to patients admitted to the burn center. Scald burns occurring from car radiator fluid comprise a significant subgroup. Although manufacturer warning labels have been placed on car radiators, these burns continue to occur. This retrospective review looks at all patients admitted to our burn center who suffered scald burns from car radiator fluid to assess the extent of this problem. During the study period, 86 patients were identified as having suffered scald burns as a result of contact with car radiator fluid. Seventy-one percent of the burn injuries occurred in the summer months. The areas most commonly burned were the head and upper extremities. Burn prevention efforts have improved greatly over the years; however, this study demonstrates that scald burns from car radiator fluid continue to cause physical, emotional, and financial devastation. The current radiator warning labels alone are not effective. The National Highway Traffic Safety Administration has proposed a new federal motor vehicle safety standard to aid in decreasing the number of scald burns from car radiators. The results of this study were submitted to the United States Department of Transportation for inclusion in a docket for federal legislation supporting these safety measures. PMID- 15353941 TI - Grease fryers: a significant danger to children. AB - Scald injuries in children often occur as the result of them pulling a container of hot liquid off a countertop or stove. To date, there have been no studies that have specifically examined the impact of the burning agent after this mechanism of injury on the magnitude of injury or the hospital resources consumed in caring for these patients. In this study, we sought to compare the extent and depth of injury, hospital resources consumed, and outcomes of children burned by pulling a container of grease or other liquid onto themselves. We retrospectively reviewed all cases of children who were admitted to our pediatric burn center from January 1995 through December 2001 who sustained scald injuries as the result of pulling a container of hot liquid onto himself or herself. Patients were evaluated for extent TBSA burned, depth of injury, the need for skin grafts, and the number of ventilator, days in the intensive care unit, and total hospital days (length of stay). We identified 196 children admitted with scald injuries caused by this mechanism. Fifty-four (27.6%) were caused by grease or oil, and 142 (72.6%) were caused by other liquids. The mean age of children burned by this mechanism was approximately 3 years, and slightly more than half were males. The mean TBSA and full-thickness burn was 18.3% and 8.1% for the grease group and 14.4% and 4.5% for the no-grease group, respectively (P <.05). Fifty-six percent of those children burned with grease required skin grafting compared with only 34% of those burned by other liquids. Twenty percent of the patients burned by grease required ventilator support compared with only 6.3% of those burned by other liquids (P <.01). Of those burned by grease, 22.2% required care in the intensive care unit compared with only 6.3% of those burned by other liquids (P <.005). Mean length of stay was 16.8 +/- 2.5 days and 9.1 +/- 1.0 days for the grease and nongrease groups, respectively (P <.001). Scalds caused by children pulling a grease fryer onto themselves result in more extensive and deeper injuries and greater consumption of hospital resources than those from other liquids. These findings support the need for safer product design and more specific warnings in product labeling and instructions as to the danger of severe scald injury to minimize the occurrence of such injuries. PMID- 15353942 TI - Milk bottle burns. AB - In recent years numerous infants have been treated at Stoke Mandeville Hospital's Burns' Unit for scalds sustained in accidents that occurred during the preparation of warm milk. These were significant burns affecting young children with susceptibly thin skin that often resulted in the injured child requiring a long stay in hospital. Frequently, formal intravenous fluid resuscitation was required as was burn excision and skin grafting. We recommend the use of milk bottle warmer to avoid all the hazards associated with use of different techniques of warming milk bottle. PMID- 15353943 TI - Pediatric and adolescent gynecology in practice and residency training. PMID- 15353944 TI - Cultural sensitivity in providing reproductive care to adolescents. AB - PURPOSE OF REVIEW: To provide an exploration of the global attitudes towards reproductive healthcare by adolescent patients, and to review key points in promoting healthy sexual development among adolescents and appropriate guidelines for communicating within the adolescent population. RECENT FINDINGS: The adolescent's perception of healthcare providers and communication with adolescents are inextricably intertwined, hence the need to address both topics. In addition, we will briefly examine the topic of female circumcision and its impact on women's sexuality. It is necessary to approach this subject because of the increase in the number of individuals that emigrate each year to the United States and Canada from countries that still practise female circumcision. Providing reproductive care for these women has considerable implications for gynecologists and other healthcare providers dealing with reproductive care issues. SUMMARY: Healthcare providers who are aware of adolescent patients' developmental stage and cultural diversity are more able to gain the trust of their patients, and consequently are more effective at addressing their needs. PMID- 15353945 TI - Vulvar and genital trauma in pediatric and adolescent gynecology. AB - PURPOSE OF REVIEW: This review will look critically at recent research articles that pertain to children and adolescents who present with genital injuries. RECENT FINDINGS: Many recently published articles have noted that the history as given by the child or adolescent is the most important factor in determining the etiology of genital injuries as abuse or accidental. The history is more important than any documented or lack of documented findings on physical examination. Distinguishing whether the injury was accidental or caused by abuse is of significance to the family and the injured child or adolescent. Genital examinations of victims of documented abuse are often normal. Examinations of pregnant adolescents may appear normal, the only evidence that intercourse has taken place being pregnancy. Documented accidental genital injuries are described; unfortunately such injuries can easily be confused with sexual abuse. Photographic documentation of injuries by colposcopic photography has allowed a new level of peer review, improving the understanding of what are normal and non specific findings, many of which were considered to be abnormal in the past. The psychological impact of living in a violent, war-torn culture, or surviving a traumatic genital injury are reviewed. New ideas on the management of genital trauma are also reported. SUMMARY: Increasingly, victims of sexual abuse are found to have normal examinations, and the victim's history becomes the most important determining factor. Peer review has led to a better understanding of the wide variations in the normal genital examination. Psychological support for these victims is important in the management of genital injuries. PMID- 15353946 TI - Adolescent sexual assault: an update of the literature. AB - PURPOSE OF REVIEW: In this review, we examine the most recent literature on adolescent sexual assault, and summarize new findings regarding prevalence, risk factors, sequelae, cultural factors, genital injury, legal issues and practice implications. RECENT FINDINGS: Child and adolescent sexual-assault victims are at risk for a range of negative outcomes, including comorbid post-traumatic stress disorder and major depressive episode, comorbid post-traumatic stress disorder and substance abuse, eating disorders, delinquency, and revictimization. Cultural factors and severity levels of trauma may serve as risk factors to such outcomes in adolescent sexual-assault victims. Compared with adults, adolescent sexual assault victims have a greater frequency of rape-related anogenital injuries, but data on healing of injuries in this population are lacking. Factors related to a child sexual-assault victim's demeanor and intelligence can influence the perceived credibility of the child as a witness to the abuse. SUMMARY: Recent studies investigating prevalence, risk factors, and sequelae of child and adolescent sexual assault highlight the need for educational programs and primary prevention interventions to educate pre-pubescent children and adolescents about sexuality, including sexual assault. In addition, further research is warranted in the area of statutory rape reporting to determine its effects on adolescent health-service-seeking behaviors and outcomes. Although most adolescent sexual assault victims do not seek acute post-rape medical care, forensic nurse examiners are often the first clinicians to encounter the adolescent sexual assault victim. Nursing protocols that standardize evidence collection as well as psychological support are important in the comprehensive care of these traumatized teens. PMID- 15353947 TI - Emergency contraceptive pills: a review of the recent literature. AB - PURPOSE OF REVIEW: The purpose of this review is to inform the reader of new information published since early 2003 about emergency contraception, with a particular focus on issues of access. RECENT FINDINGS: Research continues to document low but increasing levels of knowledge about emergency contraception, increasing use, and more positive attitudes towards emergency contraception by both patients and healthcare providers. Additional information is available about efficacy and mechanisms of action. More reports of side-effects have been published, as have studies relating to the impact of emergency contraception on sexual and contracepting behaviors. Advance provision, provision by pharmacists, and over-the-counter status have been studied as ways to improve access to emergency contraception. SUMMARY: Knowledge about the efficacy, safety, types and use of emergency contraception continues to increase. Although patients have greater awareness of and more access to emergency contraception, there are still numerous barriers to its use even in countries where it is available over the counter. Healthcare providers must continue to educate themselves and their patients about emergency contraception even when it becomes available over the counter. In countries where emergency contraception is only available by prescription, providers should offer an advance prescription or supply (where available), and use newer dosing regimens for levonorgestrel-only emergency contraception to increase adherence and efficacy. Developing collaborative practice agreements with pharmacists to increase access is also recommended. Patients should be counseled to seek follow-up if no menses occurs within 3 weeks of taking emergency contraception or if symptoms such as lower abdominal pain occur after the use of emergency contraception. PMID- 15353948 TI - Stem cell therapy for urinary incontinence and pelvic floor disorders: a novel approach. PMID- 15353949 TI - Tension-free vaginal tape procedures in women with stress urinary incontinence with and without co-existing genital prolapse. AB - PURPOSE OF REVIEW: Although the tension-free vaginal tape procedure has emerged as a feasible and effective alternative for a majority of urinary stress incontinence patients, the adaptation of the technique for genital prolapse has recently attracted much attention. This review focuses on the development of the tension-free vaginal tape procedure performed with transvaginal pelvic reconstructive surgery. RECENT FINDING: Articles since 1 April 2003 with older articles included for historical purposes were focused. Tension-free vaginal tape procedures used in prolapse surgery have mainly been performed according to the original technique. Adjustment of the vaginal tape is carried out after completion of the necessary prolapse procedures. The objective and subjective cure rate for urinary incontinence after tension-free vaginal tape with concurrent prolapse procedures is between 84.9 and 94%, and 67.3 and 88.6%, respectively. The mean complication rate, bladder perforation rate, transient urinary retention rate, period of catheterization and postoperative hospital stay are between 2.7 and 34%, 0 and 13%, 9 and 43%, 3.7 and 5.1 days, and 3.4 and 6.9 days, respectively. The implanted vaginal tape seems to 'give way' to the urethra and causes no urinary obstruction at least in studies with a short follow-up period. SUMMARY: The tension-free vaginal tape procedure performed with concurrent pelvic relaxation surgery appears to be a safe and effective treatment for urinary stress incontinence with co-existing vaginal pelvic prolapse. The implanted vaginal tape is prolapsed with the adjacent pelvic tissue, but the possibility of voiding dysfunction needs to be observed in a longer follow-up period. PMID- 15353950 TI - Anabolic effects of androgens on muscles of female pelvic floor and lower urinary tract. AB - PURPOSE OF REVIEW: This review discusses recently published data concerning the anabolic effects of androgens on muscle and the mechanism by which testosterone regulates body composition with special emphasis on the anabolic effects of androgens on the muscles of the pelvic floor and lower urinary tract. RECENT FINDINGS: Androgens have direct anabolic effects on skeletal muscle. Testosterone increases lean body mass and decreases fat mass in a dose- and concentration dependent fashion. The action of testosterone on muscle involves multiple mechanisms, including its effects on inducing protein synthesis, recruiting satellite cells, and modulating the commitment of pluripotent mesenchymal cells to myogenic lineage. Levator ani and other muscles of the pelvic floor and lower urinary tract are sensitive to the anabolic effects of testosterone. Androgen receptors are also expressed in the pelvic floor and lower urinary tract of both animals and humans. Anabolic effects of androgens may play an important role in the female pelvic-floor and lower-urinary-tract disorders. Furthermore, the interactions between androgen and nitric oxide synthase and arginase have been demonstrated, suggesting that androgens may also participate in modulating the physiological functions of lower urinary tract through nitric oxide. SUMMARY: Androgens induce muscle hypertrophy and reduce fat mass. The action of androgens in the lower urinary tract and pelvic floor is complex and may depend on their anabolic effects, hormonal modulation, receptor expression, interaction with nitric oxide synthase, or a combination of these effects. Further studies are needed to determine the precise role of androgens in women with urinary incontinence and pelvic organ prolapse. PMID- 15353951 TI - Role of three-dimensional ultrasound in assessment of women undergoing urethral bulking agent therapy. AB - PURPOSE OF REVIEW: The technique of three-dimensional ultrasound imaging has been developed over the past 15 years, and has been particularly embraced by the field of obstetrics. More recently, advances have been made in gynecological applications, with promise demonstrated in pelvic floor and lower urinary tract imaging. A clinically useful extension of three-dimensional ultrasound imaging of the lower urinary tract is the assessment of urethral bulking agent therapy, as three-dimensional ultrasound imaging provides an objective means of assessing technical outcomes (i.e. periurethral position, configuration and volume of bulking agent). Our aim is to review recent developments in the use of three dimensional ultrasound imaging in female incontinence, focusing on its role in the assessment of women undergoing urethral bulking agent therapy for stress urinary incontinence. RECENT FINDINGS: Three-dimensional ultrasound imaging provides reproducible, affordable and clinically relevant information in the assessment of urethral bulking agent therapy with collagen. Serial, long-term three-dimensional ultrasound imaging assessment has enabled a definition of the technical outcome associated with a durable improvement in continence. The optimal volume varies widely between patients, but a circumferential or horseshoe shaped periurethral configuration of collagen is strongly correlated with a durable improvement in continence. This finding is in agreement with limited data from a two-dimensional ultrasound assessment of Macroplastique. SUMMARY: Three dimensional ultrasound imaging provides an objective means of assessing technical outcomes from urethral bulking agent therapy not available previously. The volume and configuration information obtained allows for rational therapeutic decision making, particularly with regard to determining the need for re-injection and recognizing the failure of an adequate therapeutic trial. PMID- 15353952 TI - Urinary diversion following radical pelvic surgery. AB - PURPOSE OF REVIEW: To describe the reconstructive techniques in urinary diversion utilized by the gynecologist after radical pelvic surgery. The radical resection of pelvic malignancies remains an important part of the armamentarium of the gynecological oncologist. Techniques in continent urinary diversion should be used to restore these women to an acceptable quality of life. Reconstruction is the surgical challenge of this and future generations of pelvic surgeons. It is imperative that pelvic surgeons begin to consider the quality of life and the functional reintegration into society of the patients we are able to salvage with these radical surgical efforts. RECENT FINDINGS: The advent of continent urostomies has significantly improved the quality of life of patients who undergo radical tumor resection involving the bladder. Such techniques prevent the need for urostomy bags, and thus all the physical and psychological ramifications associated with them. We will review recent literature associated with the complications and the management of these complications. SUMMARY: Familiarity with techniques in continent urinary diversion by the gynecologist is an important part of the management of patients with gynecological malignancies and permanent bladder atony. PMID- 15353953 TI - Physiologic role of nitric oxide and nitric oxide synthase in female lower urinary tract. AB - PURPOSE OF REVIEW: In recent years nitric oxide (NO) has gained increasing recognition as an important neurotransmitter and cell signaling molecule with a broad range of functions in the lower urinary tract. This review discusses recently published data related to the physiologic and pathophysiologic roles of NO and nitric oxide synthase (NOS) in the female lower urinary tract. RECENT FINDINGS: Expression of three isoforms of NOS, namely endothelial NOS, neuronal NOS and inducible NOS, has been identified in various tissues of the lower urinary tract in animals and humans. In addition to its relaxation effects on bladder and urethra, NO also serves as a neurotransmitter in the lower urinary tract. The physiologic roles of NO in overactive bladder, bladder outlet obstruction, diabetic cystopathy, interstitial cystitis, and bladder inflammation have been demonstrated. SUMMARY: NO plays an important role in the micturition process and in disorders of the lower urinary tract. Improved understanding of the pathophysiologic role of NO/NOS system and of the L-arginine-NO-cGMP pathway may allow us to identify suitable therapeutic targets for lower urinary tract disorders. However, there is a need for further investigation to determine the precise function of NO in human lower urinary tract because most work thus far has been done in animal models. PMID- 15353954 TI - Typical and subtle atypical presentations of endometriosis. AB - PURPOSE OF REVIEW: To review the concept of typical, subtle and invisible endometriosis and analyze the evolution and progression of the disease, as well as some factors possibly involved in the pathogenesis. RECENT FINDINGS: Although rejected by Redwine, the concept of non-visible endometriosis has been proven to exist. Some new ideas on the implication of reactive oxygen species in the development of endometriosis and its early stages are described here. SUMMARY: The increased diagnosis of endometriosis can be explained not only by the increased experience and ability of the surgeon to detect typical and non-typical endometriotic lesions, but also by a better understanding of the pathogenesis. PMID- 15353955 TI - Bibliography. Current world literature. Adolescent and pediatric gynecology. Urogynecology. PMID- 15353957 TI - Emerging infectious disease issues in international adoptions: severe acute respiratory syndrome (SARS), avian influenza and measles. AB - PURPOSE OF REVIEW: New emerging infections over the last few years demonstrate the potential for the introduction of epidemic illness through global migration. The increasing number of children adopted internationally (>20,000 in 2003, from the United States State Department) provides a unique situation for the spread of emerging infections through the combination of international travel by parents through areas where such infections may be contracted and the nature of the living conditions for many of the orphans being placed by this process. RECENT FINDINGS: The recent literature on three emerging infections--avian influenza, severe acute respiratory syndrome (SARS) and measles--describes clinical aspects of the illnesses and their epidemiology. For avian influenza aspects of the agrarian economy in southeast Asia enabled the virus to reach the human population. The potential for further adaptation to people could set the stage for a new pandemic. SARS evolved in rural China and spread worldwide in one season with an approximate 10% mortality. Attention to public-health measures led to control of this new illness. Most recently, outbreaks of measles in Chinese orphanages have been documented. These findings demonstrate the potential of such infections to be transmitted during the process of international adoption, and in the case of measles the realization of this potential in recent reported cases from Chinese orphanages brought to the United States on commercial airlines. SUMMARY: Clinicians involved in international adoption and public-health officials assessing emerging infections need to work together in monitoring these issues. PMID- 15353958 TI - Recent tuberculosis advances in Latin America. AB - PURPOSE OF REVIEW: Tuberculosis kills more people than any other infection. Despite advances in diagnostic methods and greater understanding of the reasons for treatment failure, tuberculosis remains common throughout Latin America. RECENT FINDINGS: The impact of HIV and multidrug resistance on tuberculosis control has been enormous. HIV-positive patients may be at 10 times greater risk of multidrug resistant tuberculosis than HIV-negative patients. Hopefully, improved diagnostic techniques will allow more rapid diagnosis of tuberculosis and new colorimetric systems are being developed that will enable expedited drug sensitivity testing. However, in alarming reports, only 58% of patients were treated with the recommended treatment regime in a Brazilian study, and dropout from treatment in parts of Bolivia was common. Many failings could be combated by rigorous education of patients and physicians. In an encouraging advance, multidrug resistant tuberculosis was successfully treated in a community-based programme, saving an estimated 90% of the cost of hospital-based treatment. An opportunity to identify treatment failure earlier is demonstrated by the finding that 2 months after the initiation of therapy, positive smears were found in only 3% of those whose treatment was successful, but 74% of those whose treatment failed. SUMMARY: The importance of inexpensive and widely available drugs to treat HIV and multidrug resistant tuberculosis in Latin America is clear. The need for rapid, affordable tests for tuberculosis diagnosis, and for easy drug sensitivity testing is also evident. Finally, improving treatment success is achievable even in the resource poor setting. PMID- 15353959 TI - Falciparum malaria: current therapeutic challenges. AB - PURPOSE OF REVIEW: Malaria remains a major cause of death in much of the world. The routine treatment of malaria is currently threatened by rising rates of drug resistance. Moreover, mortality among children with severe and complicated malaria remains unacceptably high. Here we review trends in antimalarial drug resistance and report on the progress of newer drugs and drug combinations. We then review some recent literature regarding the pathological processes involved in the aetiology of severe malaria that may lead to improvements in the management of children with severe disease. RECENT FINDINGS: Resistance to first line therapies, including chloroquine and sulphadoxine/pyramethamine, continues to rise in many parts of the world. The availability of newer and more effective drugs and fixed drug combinations is hampered by financial and political considerations. Nevertheless, a number of promising drugs and supportive treatments for both mild and severe malaria are at various stages of development. SUMMARY: A range of newer drugs and fixed drug combinations are now available that are safe and effective. However, these drugs remain expensive and their introduction will require political and financial support at every level. Considerable work is still required to achieve a better understanding of the processes involved in the pathogenesis of severe and complicated malaria. Only then will it be possible to develop new and appropriate therapies that will be widely applicable. PMID- 15353960 TI - Differential diagnosis of West Nile encephalitis. AB - PURPOSE OF REVIEW: This article reviews recent developments in West Nile encephalitis. Because of the large number of individuals infected in the United States, an expanded spectrum of the disease has been recognized. Flaccid paralysis presenting as poliomyelitis-like syndrome is being increasingly recognized. RECENT FINDINGS: Since 1999, West Nile encephalitis in the United States has involved thousands of patients providing an opportunity to observe the protean manifestations of the virus. Recently, ophthalmological manifestations have been described that appear to be common and specific for the virus. Clinicians in endemic areas should be careful to distinguish between West Nile encephalitis and its mimics. The virus may occur in patients with underlying disorders that have encephalopathy as a clinical feature, and clinicians should test for the virus during the mosquito season, even in patients that appear to have an explanation for their encephalopathy. West Nile encephalitis may present as viral aseptic meningitis, meningoencephalitis, or encephalitis. Muscle weakness may or may not accompany any of these clinical variants. This virus may be transmitted via blood transfusion. SUMMARY: Clinical manifestations of West Nile encephalitis continue to expand following each year's outbreaks. New neurologic and ophthalmologic manifestations continue to be described. Because of the protean manifestations, testing should be carried out during mosquito season, even in patients that have another explanation for their encephalopathy. There is no effective therapy. Flaccid paralysis may be prolonged/permanent. Prognosis may be related to the degree of relative lymphopenia on presentation, the degree of elevation of serum ferritin levels and advanced age. The course of West Nile encephalitis and its clinical manifestations are the same in normal and compromised hosts. PMID- 15353961 TI - Hookworm infection: new developments and prospects for control. AB - PURPOSE OF REVIEW: Hookworm infection remains a major health burden in developing countries. Successful control will likely be achieved through continued advances in our understanding of the epidemiology, molecular biology and immunopathogenesis of hookworm infection. This review summarizes recent advances in each of these fields, and discusses ongoing efforts to develop vaccines against hookworm anemia and growth delay. RECENT DEVELOPMENTS: Revised estimates indicate that hookworms afflict over 700 million persons in tropical and subtropical regions. Prevalence and intensity often vary considerably at both the regional and local levels, and may be influenced by climate, soil composition, education, and socioeconomic status. Immunoepidemiological studies suggest that hookworm infection likely induces a complex mixture of host-protective and pathological immune responses. There has been substantial progress in elucidating the molecular pathogenesis of hookworm disease, primarily through the identification of a number of parasite virulence factors. Mass chemotherapy remains a mainstay of hookworm control strategies although continued use of benzimidazole anthelminthics is perhaps contributing to the development of anthelminthic resistance. Consequently, there remains a need for innovative approaches, including the development of vaccines and new chemotherapeutic agents, in order to provide effective global control of hookworm disease. SUMMARY: Hookworm infection and disease is a significant threat to global health. Recent advances, particularly those at the molecular level, have provided a wealth of opportunities to better understand pathogenesis. This will likely allow for the development of novel measures such as vaccines to complement existing control methods. PMID- 15353962 TI - Hantavirus pulmonary syndrome in Latin America. AB - PURPOSE OF REVIEW: Since the identification of hantavirus pulmonary syndrome in the United States in 1993, Latin America has been the region of the world where more hantaviruses have been identified, associated with human disease or not. This update shows the advances in the study of hantaviruses in several countries of the region and the problems that are still unresolved. RECENT FINDINGS: Clinical findings in Chile have shown differences in the classical description of hantavirus pulmonary syndrome in the United States. Hemorrhage was observed in 64% of the cases and alteration in renal function in 48%. The classification of cases in mild, self-limited forms and severe forms has a prognostic value, with statistically significant differences between survivors and nonsurvivors. Epidemiological studies have shown noticeable differences in seroprevalence of antibodies against hantaviruses in humans, ranging from about 1% to more than 40% according to geographical and ethnical differences. Risk factors continue to be related to rural activities and peridomestic sites. Rodent studies have allowed the identification of putative reservoirs of hantaviruses in Uruguay, Brazil and Argentina and detected antibodies in several rodent species not previously reported in Paraguay and Panama. An Andes virus gene-based hantavirus pulmonary syndrome vaccine is currently under study. SUMMARY: Progress in research on hantaviruses has been continuous but slow. The complex picture of the viruses, the rodent reservoirs and the clinical forms of the disease mean researchers are faced with the great challenge of properly clarifying the genetic and pathogenic relationships between hantaviruses in the Americas. PMID- 15353963 TI - Rickettsial infections--a threat to travellers? AB - PURPOSE OF REVIEW: Recent developments in cell-culture techniques and molecular methods have led to the description of several new rickettsial diseases. An update on these new infections should be of interest to health workers with patients who are international travellers. RECENT FINDINGS: Epidemic typhus was reported last year in the United States when an outbreak of murine typhus was recorded in Hawaii. Among spotted fever group rickettsioses, African tick bite fever is now probably the most common rickettsial infection in Africa with numerous cases also reported in international travellers. For the first time the Astrakhan fever rickettsia has been described outside Europe, in a French patient returning from Chad. Similarly, the first case of Rickettsia sibirica mongolotimonae infection in Africa was reported in 2004. Finally, a newly recognized agent of a spotted fever rickettsiosis, Rickettsia parkeri, has been reported in the United States during 2004. SUMMARY: Because results of serological testing are only presumptive, sophisticated methods are crucial for the diagnosis and description of new rickettsial diseases, especially in atypical cases. Modern diagnostic tools include cross-adsorption assays, Western blot testing, and cell-culture and molecular-biological methods. PMID- 15353964 TI - Morbidity in schistosomiasis: an update. AB - PURPOSE OF REVIEW: Schistosomiasis is an important poverty-related health problem and more than 200 million people are infected. This review summarizes papers from April 2003 to June 2004 with a focus on schistosomiasis morbidity and the various factors that affect the level of morbidity in endemic populations. The aim is to provide an update on the current state of knowledge and, hopefully, thereby stimulate continued research interest in this important area. RECENT FINDINGS: Research into the immune responses associated with severe morbidity has provided new insights into the mechanisms of immune regulation as well as the role of genetic predisposition to periportal fibrosis. Malaria and schistosomiasis are co endemic and co-infection with malaria may increase the level of morbidity in hepatosplenic schistosomiasis, and alter the host immune response towards schistosome antigens. Schistosome infections may render the host more susceptible to human immunodeficiency virus infection by either interfering with immune responses or increasing the risk of transmission due to genital lesions. An important advance in schistosomiasis research, and parasite genomics, is the recent availability of two major Schistosoma mansoni and Schistosoma japonicum DNA bioinformatic resources. SUMMARY: Significant advances have been achieved in our understanding of the epidemiology, immunology and genetics of schistosomiasis, and the various factors that may influence morbidity. However, good research is vital for sustainable disease control, and continued progress requires a critical mass of researchers with a range of expertise from basic parasite biology to public-health interventions. It is therefore important to strengthen research capacity in endemic countries. PMID- 15353965 TI - Advances in hepatitis B and C. AB - PURPOSE OF REVIEW: Hepatitis B and C infections are prevalent around the world and a major health burden due to the associated complications of hepatic fibrosis, cirrhosis and hepatocellular carcinoma which occur in the context of chronic infection. Significant advances are being made in assessing and treating infected patients and recent studies are now targeting patients who have failed to respond to previous treatments or who have associated co-morbidities. The purpose of this article to review the recent literature on the subject of hepatitis B and C infections with particular focus on new treatment options, new approaches in patients who have previously failed therapy and in those who have co-morbidity. RECENT FINDINGS: A large number of studies have been carried out investigating the roles of varying doses, targeting treatment in particular groups and new treatment options in patients infected with hepatitis B and C. Several key findings such as the value of prolonging treatment in patients with genotype 1 hepatitis C infection, the use of pegylated interferon in chronic hepatitis B infection and the emergence of new treatments such as adefovir for resistant hepatitis B infection, as well as treatment of patients co-infected with hepatitis C and human immunodeficiency virus, have dominated the recent literature. Patients in particular groups such as those who have had liver transplantation or who are immunosuppressed have also received added attention. SUMMARY: Hepatitis B and C infections are the focus of much current attention with particular regard to new and emerging treatment options which are becoming increasingly focused on varying patient groups. PMID- 15353966 TI - A review of viral gastroenteritis. AB - PURPOSE OF REVIEW: Since Kapakian first identified a virus in the stool of a patient with diarrhoea in 1972, many viruses have been described that cause diarrhoea directly or indirectly. It is now appreciated that viruses are the most common cause of diarrhoeal illness worldwide. Although bacteria and other pathogens cause significant numbers of gastroenteritis, it is the viruses that are dealt with in this review. The viruses responsible will be discussed individually. RECENT FINDINGS: Rotavirus remains the leading cause of diarrhoeal disease overall, with the newly designated calicivirus family causing the most outbreaks in the industrialized nations. As diagnostic techniques improve, however, the importance of astrovirus and other previously under-reported pathogens is becoming more apparent and the number of viruses associated with gastroenteritis continues to increase. The emergence of severe acute respiratory syndrome coronavirus, arguably the most important emerging infection of recent years and a cause of significant gastrointestinal disease, is also discussed. SUMMARY: No effective treatments have been developed for viral gastroenteritis. Current efforts are targeted at the development of suitable vaccines and the implementation of infection control measures. PMID- 15353967 TI - Human norovirus infection and the lessons from animal caliciviruses. AB - PURPOSE OF REVIEW: Human noroviruses are a major cause of infectious intestinal disease, particularly in the health sector, with considerable knock-on effects on care provision through ward closures and staff sickness. This review will describe recent advances in our understanding of human noroviruses. In addition, we will consider related nonhuman caliciviruses to highlight some potential difficulties in the control of caliciviral disease. RECENT FINDINGS: Using more sensitive reverse transcriptase polymerase chain reaction based assays, noroviruses are now recognized as the most common cause of infectious intestinal disease in the community, as well as outbreaks of the infectious intestinal disease. After recovery from acute disease, some individuals continue shedding norovirus, particularly if immunosuppressed. The noroviruses are extremely variable, which has important implications for protection following challenge, and for future vaccination. From amongst this variability, new strains have emerged with the potential to spread widely. Recently a mouse norovirus has been identified which will afford new insights into the biology of these important viruses. Studies on human susceptibility have identified some resistant individuals in the population and a potential virus receptor, which may lead to the development of novel antiviral therapies. SUMMARY: Lack of cell culture systems for the human noroviruses is being overcome by molecular technologies. Such studies have provided new insight into the significance and epidemiology of these viruses and opened the possibility of disease control through vaccination. Work on nonhuman caliciviruses has interesting parallels with human noroviruses, and provides new insights into the understanding of these important human pathogens. PMID- 15353968 TI - Potential therapeutic agents for the prevention and treatment of haemolytic uraemic syndrome in shiga toxin producing Escherichia coli infection. AB - PURPOSE OF REVIEW: Shiga toxin producing Escherichia coli (STEC) cause a wide spectrum of disease ranging from asymptomatic carriage through to haemorrhagic colitis and the haemolytic uraemic syndrome. There are no current therapeutic interventions available in clinical practice that can prevent the development of haemolytic uraemic syndrome. A number of newly developed agents offer the potential for the treatment of STEC-associated disease. RECENT FINDINGS: Three different classes of agent designed to bind and inactivate shiga toxin have now been developed. Synthetic toxin binders, recombinant bacteria and monoclonal antibodies provide potentially potent agents that could prevent the development of haemolytic uraemic syndrome. These agents have been shown in animal models of STEC disease to be effective. A recent clinical trial of one synthetic toxin binder showed no benefit in established haemolytic uraemic syndrome. More potent toxin binders, however, have since been developed and await human clinical trials. It is likely to be important that these agents are administered early in the course of disease in order to have maximum efficacy. Although rapid diagnostic techniques are available for the diagnosis of STEC disease, they still rely on stool culture. SUMMARY: Clinicians need to maintain a high level of suspicion of STEC disease as the diagnosis is often made on epidemiological and clinical grounds. This will allow potential cases to be identified early and treated appropriately. PMID- 15353969 TI - Cryptosporidiosis: an update in molecular epidemiology. AB - PURPOSE OF REVIEW: Molecular tools have been developed to detect and differentiate Cryptosporidium at the species/genotype and subtype levels. These tools have been increasingly used in the characterization of the transmission of Cryptosporidium spp. This review addresses the most recent developments in molecular epidemiology of cryptosporidiosis. RECENT FINDINGS: The recent development of subtyping tools has led to better understanding of the population genetics and transmission of Cryptosporidium in humans. The population structure of C. parvum and C. hominis is apparently more complicated than previously suggested, with the likely existence of both clonal and panmictic populations. Thus, the transmission of C. parvum (genotype II) in humans is shown to be different in different areas, with zoonotic transmission important in certain places and anthroponotic transmission in others. The use of molecular tools has also led to the identification of geographic and temporal differences in the transmission of C. parvum and C. hominis, and better appreciation of the public health importance of other Cryptosporidium species/genotypes and the frequency of infections with mixed genotypes or subtypes. SUMMARY: Factors involved in the transmission of human cryptosporidiosis are difficult to examine using conventional methods. The use of molecular tools has been helpful in the assessment of the zoonotic potential of various Cryptosporidium spp. and sources of human infections, and has started to play a significant role in the characterization of transmission dynamic in endemic and epidemic areas. PMID- 15353970 TI - Bibliography. Current world literature. Tropical and travel-associated diseases. PMID- 15353972 TI - Monitoring HIV/AIDS in Europe's migrant communities and ethnic minorities. PMID- 15353973 TI - HLA- and dose-dependent immunogenicity of a peptide-based HIV-1 immunotherapy candidate (Vacc-4x). AB - OBJECTIVE: The Vacc-4x immunotherapy candidate is composed of four modified peptides corresponding to conserved domains of the HIV-1 protein p24 that preferentially include HLA-A2 restricted elements. Dose-dependent safety and immunogenicity of Vacc-4x and the significance of a HLA-A2 haplotype were examined. DESIGN: Non-AIDS, HIV-1 infected healthy patients (n = 40) stable on HAART with CD4 counts > 300 x 10 cells/l were randomized to receive either low dose or high-dose Vacc-4x over 26 weeks in an open, prospective phase II clinical trial. METHODS: Patients received a total of 10 intradermal injections, using recombinant granulocyte-macrophage colony stimulating factor as a local adjuvant. Vacc-4x-specific cellular responses were monitored in vivo by delayed-type hypersensitivity (DTH) skin test infiltrates and in vitro by both T-cell proliferation, and induction /secretion of cytokines. RESULTS: Most patients developed Vacc-4x-specific DTHs (90%) and proliferative T-cell responses (80%) that were inter-related in magnitude. High-dose Vacc-4x generally induced stronger specific immune responses than low dose in terms of DTH areas and CD4 and CD8 T-cell proliferation. Only HLA-A2 negative patients had a definite dose advantage, and this subgroup had in fact the best overall DTH and proliferative responses. In contrast, no significant dose difference was observed for HLA-A2 positive patients. No serious adverse events were reported. CONCLUSIONS: HIV associated specific responses were safely induced in most patients by Vacc-4x in a dose-dependent manner and were also influenced by the HLA haplotype. PMID- 15353974 TI - Pre-seroconversion immune status predicts the rate of CD4 T cell decline following HIV infection. AB - OBJECTIVE: To study whether immune status prior to HIV seroconversion predicts CD4 T cell decline during HIV infection. DESIGN: Prospective cohort study including 51 injecting drug users (IDU) who were HIV negative at study entry and seroconverted for HIV during follow-up. METHODS: Cryopreserved peripheral blood mononuclear cells obtained before HIV seroconversion were used to measure naive (CD45RO-CD27+), memory (CD45RO+CD27+), and total CD4 T cell numbers, the fraction of dividing Ki67+CD4+ T cells, and CD4 T cell receptor excision circles (TREC). The effect of pre-seroconversion immune status, as defined by these markers, on the rate of CD4 T cell decline during HIV infection was assessed using linear regression for repeated measurements. RESULTS: IDU with low pre-seroconversion CD4 T cell TREC contents lost CD4 T cells at a significantly faster rate during HIV infection than those with a high CD4 T cell TREC content. IDU with higher pre seroconversion CD4 T cell numbers had a significantly steeper CD4 T cell decline in the first 3 months of HIV infection, but their CD4 T cell counts remained higher throughout HIV infection. Intermediate levels of pre-seroconversion dividing Ki67+CD4+ T cells were associated with a significantly steeper CD4 cell decline than high levels. IDU with the highest pre-seroconversion drug-injecting frequencies showed slower CD4 T cell decline than those who injected less. No correlation was present between pre-seroconversion immune markers and the pre seroconversion duration or intensity of drug use. CONCLUSION: Among IDU, immune status prior to HIV infection as measured by TREC content affects the disease course after HIV seroconversion. PMID- 15353975 TI - The impact of initial highly active antiretroviral therapy on future treatment sequences in HIV infection. AB - OBJECTIVES: To determine whether the initial use of non-nucleoside reverse transcriptase inhibitors (NNRTI) or protease inhibitors (PI) differentially influences subsequent HIV therapy. DESIGN: A cohort study using a prospective clinical database in a university-based HIV clinic. SUBJECTS: A total of 440 HIV seropositive patients, naive or nucleoside experienced, initiating therapy with either an NNRTI or PI between January 1998 and July 2003 and followed to December 2003. MAIN OUTCOME MEASURES: Time until stopping the first regimen and until exposure to all antiretroviral classes (excluding tenofovir and enfuvirtide) according to the type of initial regimen. RESULTS: A total of 291 subjects initiated HAART with PI and 149 with NNRTI; median follow-up 3.1 and 2.3 years, respectively. Subjects starting NNRTI remained on their initial regimens longer (median time to change 2.1 versus 1.6 years; log rank P = 0.03). Overall, subjects initiating NNRTI-based regimens were less likely to alter their therapy. Previous nucleoside exposure was an important predictor of treatment modification. Subjects initiating NNRTI-based HAART were also less likely to experience virological failure than those initiating PI-based HAART. Individuals starting with NNRTI were exposed to fewer regimens (15 versus 25% received three or fewer regimens), and showed a trend towards lower rates of three-class exposure (7 versus 12%). CONCLUSION: There is a high rate of treatment modification among patients initiating HAART. The initial use of NNRTI-based HAART was associated with more durable treatment and lower rates of virological failure, which may translate into a reduced need for multiple salvage therapies. PMID- 15353976 TI - Highly active antiretroviral therapies among HIV-1-infected children in Abidjan, Cote d'Ivoire. AB - OBJECTIVE: To describe the effect of highly active antiretroviral therapy (HAART) in HIV-1-infected African children. STUDY DESIGN: Observational ANRS 1244 cohort of 159 children with HIV between October 2000 and September 2002; 78 children (49%) receiving HAART were followed for a mean duration of 21 months. METHODS: Weight-for-age Z-scores (WAZ), height-for-age Z-scores (HAZ), CD4 lymphocyte count and HIV-1 RNA viral load were measured before initiating HAART and every 6 months during treatment. Probability of survival and incidences of pneumonia and acute diarrhoea were calculated. RESULTS: Values before and after 620 days of HAART, respectively, were -2.02 and -1.39 for mean WAZ, (P < 0.01); -2.03 and 1.83 for mean HAZ (P = 0.51); 0.07 and 0.025/child-month (P = 0.002) for incidence of pneumonia; and 0.12 and 0.048/child-month for incidence of acute diarrhoea (P < 0.001) (incidence changes statistically significant only in children < 6.5 years). Overall, the probability of survival under HAART was 72.8% at 24 months for children with < 5% CD4 cells versus 97.8% in children with >/= 5% (P < 0.01). At HAART initiation, median viral load and CD4 cell percentage were 5.41 log10 copies/ml and 7.7%, respectively. After 756 days of HAART, on average, 50% of patients had undetectable viral load and 10% had 2.4-3.0 log10 copies/ml. The median CD4 percentage was 22.5%. CONCLUSION: In resource-limited setting, it is possible to use HAART to treat African children. This treatment appears as effective as in developed countries. PMID- 15353977 TI - Response to highly active antiretroviral therapy varies with age: the UK and Ireland Collaborative HIV Paediatric Study. AB - OBJECTIVE: To evaluate the effect of age, CD4 percentage (CD4%) and plasma HIV-1 RNA on response to highly active antiretroviral therapy (HAART) in previously untreated children. DESIGN: Cohort study. METHODS: We examined the association between age at HAART initiation, and CD4 and HIV-1 RNA response using logistic and Cox regression, adjusting for sex, route of infection and pre-HAART values. RESULTS: CD4% increases of > 10% at 6 months were more likely in younger children [odds ratio (OR), 0.84 per year, P < 0.001] and those with lower pre-HAART CD4% (OR, 0.67 per 5% higher, P < 0.001), but were not related to pre-HAART HIV-1 RNA (P = 0.6). In contrast, HIV-1 RNA suppression < 400 copies/ml at 6 months was more likely in older children (OR, 1.09 per year, P = 0.03), and was unrelated to pre-HAART HIV-1 RNA or CD4% (P > 0.3). CD4% was still increasing during the second year following HAART initiation (60% followed > 24 months). Longer-term increases in CD4% occurred faster, and decreases in HIV-1 RNA occurred more slowly in younger children. The median time to CD4% >/= 30% after initiating HAART with CD4% 28 days after HIV diagnosis) and subsequent mortality. Patients were grouped by CD4 cell count into those with < 200, 200-500 and > 500 x 10 cells/l. RESULTS: Incident TB was diagnosed among 159 of 2012 patients, with 4973 person-years of observation time. In 105/159 (66.0%), the diagnosis was confirmed by direct microscopy or culture. Incidence of TB was highest in the group with < 200 x 10 cells/l (9.1/100 and 8.8/100 person-years in HIV-1 and HIV-2, respectively). Adjusted for CD4 cell count, there was no significant difference in incidence or mortality following TB between HIV-1- and HIV-2-infected patients. Mortality rate was higher in those with incident TB and HIV infection, most markedly in the group with the highest CD4 cell count (hazard ratio, 10.0; 95% confidence interval, 5.1-19.7). CONCLUSION: Adjusted for CD4 cell count, incidence of TB was similar among HIV-1- and HIV-2-infected patients. Mortality rates after TB diagnosis were similar in both groups and high compared with those without TB. PMID- 15353980 TI - Perceived viral load, but not actual HIV-1-RNA load, is associated with sexual risk behaviour among HIV-infected homosexual men. AB - BACKGROUND: Increases in sexual risk behaviour and sexually transmitted infections among HIV-infected homosexual men after the introduction of highly active antiretroviral therapy (HAART) confirm the need for innovative prevention activities. The present study focused on time trends in sexual risk behaviour and predictors for unprotected anal intercourse in the HAART era among HIV-infected homosexual men. METHODS: In 2000-2003, 57 HIV-infected homosexual men (mean age 45 years) were interviewed in three serial data waves. Logistic regression, correcting for repeated measurements, was used to assess time trends in risky sex, and the association between HAART-related beliefs, and both the perceived and actual viral load level and CD4 cell counts and subsequent risky sex. RESULTS: Risky sex with casual partners increased from 10.5% in 2000 to 27.8% in 2003 (P < 0.01), and with steady partners of negative or unknown HIV status from 5.3% to 10.7% (P = 0.6). Homosexual men with a favourable perception of their viral load were more likely to engage in subsequent risky sex with steady partners of negative or unknown HIV status than men with a less favourable perception of their viral load; this association was independent of the actual HIV-1-RNA load and CD4 cell counts. CONCLUSION: Risky sex increased in this group of HIV-infected homosexual men. The perceived viral load, but not the actual load, is associated with subsequent risky sex with steady partners of negative or unknown HIV status. Care givers should discuss with patients not only their actual viral load and CD4 cell count but also their perceived viral load. PMID- 15353981 TI - Proviral HIV-DNA predicts viral rebound and viral setpoint after structured treatment interruptions. AB - In HIV-1-infected patients with long-term undetectable viraemia on highly active antiretroviral treatment (HAART), we found that pre-HAART plasma viraemia and the baseline proviral DNA level were significantly associated with the viraemia setpoint during scheduled treatment interruptions. In long-term treated patients, pre-HAART viraemia may not be available, and in these circumstances proviral DNA, measured at the time of scheduled treatment interruption, can help to identify patients likely to reach a low viraemia setpoint after treatment interruption. PMID- 15353982 TI - Long-term changes in circulating CD4 T lymphocytes in virologically suppressed patients after 6 years of highly active antiretroviral therapy. AB - Highly active antiretroviral therapy (HAART) initiated in advanced HIV disease is associated with CD4 lymphocyte increases (200-300 cells/mm3 after 2-4 years), although longer-term cellular dynamics have not been studied. We observed a significant median CD4 lymphocyte increase of 126 cells/mm3 and 54 naive CD4 lymphocytes from year 3 to 6 of HAART among 20 individuals with pre-HAART CD4 cell counts of 100-300 cells/mm3. This cohort represents the longest prospective immunological follow-up of virologically suppressed patients on HAART. PMID- 15353983 TI - Adipose tissue expression of IL-18 and HIV-associated lipodystrophy. AB - IL-18 is an inducer of apoptosis/tissue injury. IL-18 messenger RNA expression was examined in adipose tissue (AT) obtained from HIV patients with lipodystrophy, without lipodystrophy and healthy controls. IL-18 mRNA was expressed in AT at increased levels in lipodystrophy-positive compared with lipodystrophy-negative patients and healthy controls. Higher levels of IL-18 mRNA were found in femoral-gluteal AT compared with abdominal AT, and correlated with limb fat loss. These findings suggest that IL-18 is linked to HIV-associated lipodystrophy. PMID- 15353984 TI - Penetration of enfuvirtide, tenofovir, efavirenz, and protease inhibitors in the genital tract of HIV-1-infected men. AB - One likely mechanism of virological failure is poor antiretroviral drug diffusion in sites of viral replication such as the genital tract. We measured antiretroviral drug concentrations in blood and semen in 13 HIV-infected men failing treatment. Enfuvirtide did not cross the blood-testis barrier, whereas tenofovir accumulated in semen. Unlike indinavir, semen concentrations of lopinavir, amprenavir, saquinavir and efavirenz were ineffective. These are worrying findings, because suboptimal semen drug concentrations may enhance the risk of sexually transmitted drug-resistant HIV variants. PMID- 15353985 TI - High incidence of atypical mycobacteriosis in African HIV-infected adults with low CD4 cell counts: a 6-year cohort study in Cote d'Ivoire. AB - The role of non-tuberculous mycobacteriosis (NTM) in HIV-related diseases in sub Saharan Africa has long been controversial. In a 6-year cohort of 721 HIV infected adults with systematic BACTEC blood cultures in Abidjan, Cote d'Ivoire, the incidence of NTM was 1.8/100 person-years overall and 12.2/100 person-years in patients with baseline CD4 cell counts < 100 cells/mm3. In sub-Saharan Africa, where most patients start highly active antiretroviral therapy with low CD4 cell counts, improving the diagnosis of NTM may be relevant. PMID- 15353986 TI - Isolated lopinavir resistance after virological rebound of a ritonavir/lopinavir based regimen. PMID- 15353987 TI - Enfuvirtide prescription at the end of pregnancy to a multi-treated HIV-infected woman with virological breakthrough. PMID- 15353988 TI - An epidemic of enterovirus 71 infection among HIV-1-infected orphans in Nairobi. PMID- 15353990 TI - Gas flow into and within the middle ear. AB - HYPOTHESIS: Both a normal and a narrowed eustachian tube (ET) are capable of equilibrating pressures between the middle ear (ME) and the atmosphere almost instantaneously. OBJECTIVES: The objective of this study was to assess experimentally the effect of narrowing a simulated ET isthmus on air passage into the ME. METHODS: A Perspex ME model (0.5 mL) was constructed in which a 1.5-mm long ET of 0.07- to 1.0-mm diameter and a "mastoid" of 0- to 10-mL volume were changeable. The ET could be opened and closed with a valve. A -5 mm H2O pressure difference between the system and the atmosphere was created by withdrawing gas from the system. The time required to equalize these pressures after opening the valve to the atmosphere were measured with a pressure transducer. RESULTS: A pressure difference of -5 mm H2O was created in the system when 1.3 to 6.5 microL of ME gas was removed. On ET valve opening, the pressure was equalized within 0.1 and 0.15 to 0.3 seconds for ET diameters of 1.00 and 0.07 mm, respectively, depending also on the "mastoid" volume. Similar results were obtained when the pressure was measured through the "tympanic isthmus" and "aditus ad antrum." CONCLUSIONS: Our model shows that under ordinary physiological conditions, the amount of gas that can pass through the ET during swallowing time (0.4 sec) is potentially higher than required to equalize a negative pressure. This is also the case when the ET is very narrow and open for a very short time. It is unlikely that any narrowing of the tube will, by itself, hamper gas transfer into or within the ME, as long as the ET is not totally obstructed. PMID- 15353991 TI - Retrograde mastoidectomy with canal wall reconstruction: a follow-up report. AB - OBJECTIVE: To evaluate long-term results of retrograde mastoidectomy with canal wall reconstruction as a single-stage technique for cholesteatoma removal. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: Forty-six patients, representing 50 ears (20 pediatric and 30 adult), who had undergone surgery for cholesteatoma removal with said technique and had an average follow-up of 7.8 years. INTERVENTIONS: Temporary removal of the upper canal wall, in association with a retrograde-type mastoidectomy, for full exposure and extirpation of the disease, followed by reconstruction of the canal defect using cymba cartilage. MAIN OUTCOME MEASURES: Preoperative and short- and long-term postoperative audiogram, obtained as four-frequency pure-tone average air-bone gap. Complications, including presence of recurrent or residual cholesteatoma, need for tube insertion, perforation, and poor hearing requiring revision surgery, were also reported and correlated with the patient's tobacco use. RESULTS: The average preoperative, short-term postoperative, and long-term postoperative pure-tone average air-bone gap was 25.6+/-11.2 dB, 11.0+/-5.7 dB, and 12.4+/-6.4 dB, respectively. There were significant differences between the pre- and postoperative values (p < 0.5), but there was no significant difference between short- and long-term hearing results. Recurrent cholesteatomas were seen in eight ears (16%); pressure-equalizing tube insertion was performed postoperatively in nine ears (18%); a perforation was seen in one ear (2%); and two ears (4%) had poor hearing results requiring second-look surgery. The long term complication rate of smokers was 79% (15 of 19), compared with 16% (5 of 31) for nonsmokers. CONCLUSION: This single-stage technique for cholesteatoma removal and canal wall reconstruction showed acceptable long-term results, but tobacco use was associated with a higher long-term complication rate. PMID- 15353992 TI - A possible role for nitric oxide in osteoclastogenesis associated with cholesteatoma. AB - HYPOTHESIS: This study was designed to investigate the potential role of nitric oxide in cholesteatoma-induced bone resorption, in vitro and in vivo. BACKGROUND: Cholesteatoma is a disease of inflammatory bone resorption in the middle ear leading to hearing loss and vestibular dysfunction. Inappropriate activation of osteoclasts causes the morbidity associated with this disease. Previous studies suggest nitric oxide may be an important mediator of osteoclast function. METHODS: A murine model of cholesteatoma induced bone resorption was used to demonstrate nitric oxide synthase (NOS) gene expression and the effect of a NOS inhibitor. An in vitro osteoclast culture method was used to demonstrate the effect of nitric oxide on isolated osteoclasts. Osteoclast development was assayed by counting the number of mature osteoclasts; activity was assayed by measuring the amount of resorbed bone. RESULTS: Quantitative reverse transcriptase-polymerase chain reaction results demonstrated the temporal expression of all three NOS isoforms in vivo. NOS I demonstrated very low levels of expressions throughout the duration of the study with no change in expression in response to keratin implant. Similarly, NOS III also demonstrated low levels of expression and no change in response to keratin. NOS II was highly upregulated in response to keratin throughout the duration of the study. In vitro, pharmacological nitric oxide donors--sodium nitroprusside and S-nitroso-N-acetyl D,L-penicillamine--dose-dependently stimulated osteoclast resorption. Alone, interferon gamma (IFNgamma)--but not IL-1beta or TNFalpha--generated nitrite in vitro. A cytokine cocktail of IL-1beta, TNFalpha, and IFNgamma synergistically enhanced nitrite production. Nitrite production was blocked by the addition of aminoguanidine (AG), suggesting that AG-inhibited cytokine mediated nitrite production. However, in an in vivo model of cholesteatoma-induced bone resorption, the osteoclast response of AG-treated mice was not statistically different from untreated controls. CONCLUSIONS: All three NOS isoforms were expressed in an in vivo model of cholesteatoma-induced bone resorption with significant upregulation of NOS II throughout the study. Exogenously administered nitric oxide dose-dependently enhanced osteoclast activation in vitro. The pro inflammatory cytokines, IL-1beta, TNFalpha, and IFNgamma, synergistically induce nitrite production, which was abrogated by treatment with the nitric oxide synthase inhibitor, AG. Although AG suppresses nitrite production in vitro, treatment had no effect on osteoclast response in vivo, suggesting that the effects of inflammatory cytokines on osteoclast response were mediated through other pathways. PMID- 15353993 TI - A new approach for malleus/incus fixation: no prosthesis necessary. AB - OBJECTIVE: To describe a novel approach to manage malleus/incus fixation. STUDY DESIGN: Retrospective review of 363 patients with conductive hearing loss operated on since 1996. SETTING: Academic tertiary referral center. PATIENTS: Three hundred sixty-three patients with conductive hearing loss, an intact tympanic membrane, and without history for chronic infection underwent middle ear exploration. Three hundred forty-three had otosclerosis and underwent laser stapedotomy; the remaining 20 patients had laser release of their malleus/incus fixation. INTERVENTION: Twenty patients are presented in this paper. Nineteen patients were diagnosed with malleus fixation before surgery. One patient had a total perforation and mobility of the ossicular chain was not verified preoperatively. This patient was, intraoperatively, noted to have complete ossicular fixation. Conductive hearing loss was identified using audiometry and tuning forks. Nineteen of the 20 patients had the diagnosis confirmed using micropneumotoscopy and noting immobility of the malleus. A transcanal approach was used, and the malleus/incus fixation was released using a laser. This space was expanded upon using a drill. A 1.5-2.0 mm space was created where the ossicular fixation existed, thereby reducing the likelihood of refixation. MAIN OUTCOME MEASURE: Audiometric studies before and after intervention were compared. One to seven years of follow up are provided. RESULTS: Preoperative air-bone gaps ranged from 18 dB to 51 dB and averaged 33 dB. Postoperative air-bone gaps ranged from 1-36 dB and averaged 13 dB. No patients have experienced refixation. There were two complications: one perforation requiring a tympanoplasty and one patient sustaining a 20 dB high-frequency sensorineural loss, most likely secondary to inadvertent drill contact to the ossicular chain. CONCLUSION: When fixation of the malleus and/or incus is found, treatment options exist. A common technique involves removal of the incus and head of the malleus and reconstruction with an incus interposition or a partial ossicular prosthesis. Another technique proposed by the senior author (M.D.S.) is maintenance of the normal anatomy and use of the potassium-titanyl-phosphate laser and drill to free the ossicles and widen the epitympanum. This series shows successful closure of the air-bone gap with this technique. PMID- 15353994 TI - Bone destruction resulting from rupture of a cholesteatoma sac: temporal bone pathology. AB - OBJECTIVE: The purpose of the current study was to research the pathogenesis of bony destruction of cholesteatoma. STUDY DESIGN: We conducted a case report. SETTING: The study was performed at Fukushima Medical University. PATIENTS: The first case involved a 21 trisomy, whereas the second case was cancer of the hypopharynx. Both cases showed cholesteatoma. RESULTS: The following histopathologic findings in the temporal bones of cholesteatoma were obtained. Bony destruction in cholesteatoma was detected in the lesion of the rupture of the cholesteatoma sac. Epithelial debris of cholesteatoma was scattered throughout the rupture of the cholesteatoma sac. Rupture of the matrix was the result of a small abscess. CONCLUSION: Rupture of the cholesteatoma sac was believed to have been a pathway of either endogenous substances from the matrix and/or epithelial debris of cholesteatoma. PMID- 15353995 TI - Success of cartilage grafting in revision tympanoplasty without mastoidectomy. AB - OBJECTIVE: Candidates for revision tympanoplasty have experienced at least one failed attempt at repair of the tympanic membrane and are, therefore, at higher risk for subsequent repair failure. The adjunctive use of mastoidectomy with tympanoplasty in those patients with noncholesteatomatous chronic otitis media is often used to decrease the risk for subsequent failure. However, at this institution, where we use cartilage tympanoplasty, mastoidectomy is rarely performed in the absence of cholesteatoma. Our objective was to assess outcomes in patients undergoing revision tympanoplasty without mastoidectomy using cartilage grafting. STUDY DESIGN: We conducted a retrospective case review. SETTING: Tertiary referral center. PATIENTS: A total of 95 patients (42 female, 53 male; 5-81 yr of age) with a recurrent perforation who were treated surgically with cartilage tympanoplasty without mastoidectomy were included in the chart review. Patients must have undergone at least one previous tympanoplasty without mastoidectomy and had to have complete audiologic and chart follow up. INTERVENTIONS: An underlay tympanoplasty technique using either a tragal cartilage-perichondrium island graft or palisaded concha cymba cartilage was used. Ossiculoplasty was performed as needed. MAIN OUTCOME MEASURE: Main outcome measures were incidence of reperforation of the grafted tympanic membrane, hearing result, and prevalence of other complications. RESULTS: Successful closure without reperforation was obtained in 90 of 95 patients (94.7%). Average postoperative pure-tone average air-bone gap was 12.2 +/-7.3 dB compared with 24.6+/-13.8 dB preoperatively (p <0.001). CONCLUSIONS: Revision tympanoplasty with cartilage provided equivalent results to tympanoplasty with mastoidectomy. Thus, mastoidectomy may not be necessary in revision tympanoplasty in the absence of cholesteatoma if the repair is made with cartilage. PMID- 15353996 TI - Limitations of titanium dioxide and aluminum oxide as ossicular replacement materials: an evaluation of the effects of porosity on ceramic prostheses. AB - BACKGROUND: Because the performance of titanium dioxide (TiO2) has not yet been assessed in the unique environment of the middle ear, its role as an ossicular replacement prototype in the form of a total ossicular replacement prosthesis (TORP) was tested and compared with aluminum oxide (Al2O3), once considered to be a suitable implant material. METHODS: Ossiculoplasty was performed by implanting TORPs into the tympanic cavities of rabbits. After an implantation period of 28, 84, or 300 days, the petrous bones were extracted, whereby the biocompatibility of the prostheses was examined using light microscopy and scanning electron microscopy to determine morphologic changes in situ. Proper implant placement and functionality was tested via manual manipulation. RESULTS: Mucosa was seen covering most of the implants by day 84. Inflammatory cells were not observed in any of the specimens examined. The macroporous TiO2 TORPs were subjected to osseous infiltration, material dissolution, and fragmentation, whereas the microporous TiO2 implants were subjected to an increasing frequency of fissure formations. The Al2O3 prostheses demonstrated signs of material dissolution by producing encapsulated aggregates during the experimental trial period. CONCLUSIONS: Neither the macroporous nor microporous oxide ceramics were able to withstand the oscillatory stress to which they were continually subjected. Although porosity allows for the rapid integration of an implant material into a biological environment, its properties are not suited to fulfill the requirements of strength and long-term stability, which are demanded of middle ear prostheses. PMID- 15353997 TI - Vein versus tragal perichondrium in stapedotomy. AB - OBJECTIVE: To assess, in otosclerosis surgery, whether the vein or the tragal perichondrium in stapedotomy with interposition yields the better long-term hearing outcome. STUDY DESIGN: A retrospective chart review of prospectively collected audiometric data of 452 ears. SETTING: Academic tertiary otology neurotology referral center. PATIENTS: Four hundred fifty-two stapedotomies with interposition were performed in 412 patients (bilateral in 40 patients) by the senior author (R.C.) between 1987 and 1998. A tragal perichondrium graft was used in 314 cases and a vein graft was used in 138 cases as sealing material of the oval window. MAIN OUTCOME MEASURES: Audiometric data were recorded at 4 months, at 1 year, and at 3 years after surgery after American Academy of Otolaryngology Head and Neck Surgery guidelines, except for thresholds at 3 kHz, which were not available and which were replaced with those at 4 kHz. RESULTS: There were no significant intergroup differences in initial or late postoperative hearing outcome with regard to change in the pure-tone average bone conduction and air bone gaps, or sensorineural hearing loss. Ears treated with a vein graft showed statistically better postoperative 2-kHz air-bone gap closure (p =0.0157), but the pure-tone average air-bone gap difference was not significant. Postoperative air-bone gap closure to within 10 dB was achieved in 91% of cases in the vein group and in 76% of cases in the perichondrium group. Specific study of the bone conduction level at 4 kHz showed a sensorineural hearing loss greater than 10 dB in 8% of cases in the vein group and in 11% of cases in the perichondrium group. One case of complete sensorineural hearing loss was observed with a tragal perichondrium graft (0.22%). CONCLUSION: These results suggest that the vein should be preferred to the tragal perichondrium in stapedotomy with interposition. PMID- 15353998 TI - Variable clinical features in patients with CDH23 mutations (USH1D-DFNB12). AB - OBJECTIVE: To describe the findings of audiovestibular and ophthalmologic examinations in four families with mutations in the CDH23 gene. STUDY DESIGN: Family study. SETTING: Tertiary referral center. PATIENTS: Four DFNB12 patients from a large consanguineous Dutch family and six patients from three different Usher syndrome Type ID families were examined. All were identified by at least one pathogenic mutation in the CDH23 gene. METHODS: Audiovestibular examinations consisted of standard pure-tone audiometry, vestibulo-ocular reflex, optokinetic nystagmus, and in some cases the cervico-ocular reflex. Linear regression analysis was used to evaluate progression of hearing impairment, and the degree of hearing impairment of DFNB12 was compared with that found for USH1D. Ophthalmologic examinations consisted of best-corrected visual acuity, Goldmann perimetry, slit-lamp examinations, color vision testing, dark adaptation, electroretinography, electro-oculography, funduscopy and photography of the retina, and sometimes fluorescein angiography. RESULTS: The USH1D patients had significantly worse hearing impairment than the DFNB12 patients. The DFNB12 patients, identified by missense mutations in CDH23, had normal retinal and vestibular function. All USH1D patients had splice-site mutations in CDH23 and a typical Usher syndrome Type I phenotype. One DFNB12 patient had slightly abnormal yellowish flecks in the posterior poles of both eyes. CONCLUSION: Recessive missense mutations in CDH23 lead to a milder phenotype (DFNB12) than splice-site mutations (USH1D); however, abnormal bilateral flecks, suggestive for lipofuscin accumulation, can be observed in DFNB12 patients. PMID- 15353999 TI - Characterization of a new mouse mutant, flouncer, with a balance defect and inner ear malformation. AB - HYPOTHESIS: Balance anomalies are often associated with abnormalities of the vestibular part of the inner ear. We studied a newly generated mouse mutant with balance defects and asked whether its behavioral anomalies were associated with inner ear defects. Furthermore, we asked whether the mutation responsible for the defects was located in the same region of mouse chromosome 4 as several other mouse mutations that we have previously described. BACKGROUND: Phenotypic and genotypic analysis of mouse mutants with hearing or balance problems has helped greatly with the identification of the genes involved in deafness and has contributed to the understanding of mechanisms of normal hearing and balance. This article describes a new mouse mutant, flouncer, that shows a balance defect. The flouncer mutation shows semidominant inheritance, and was generated by mutagenesis using N- ethyl-N- nitrosourea. METHODS: Hearing was assessed by the Preyer reflex (ear-flick) test. Behavioral tests including open field and swimming tests were performed. The morphology of the middle and inner ears was investigated by microdissection, clearing using glycerol, paint-filling of the labyrinth, and scanning electron microscopy. RESULTS: Flouncer mutants showed vestibular dysfunction but do respond to sounds. Phenotypically, mutants had various degrees of truncation of the lateral semicircular canals, small or obliterated round window of the cochlea, and mild morphologic anomalies of the stapes. Flouncer mutants showed circling behavior and hyperactivity. Linkage mapping using a backcross has indicated that the mutation lies in proximal chromosome 4 proximal to D4Mit171. CONCLUSION: The lateral semicircular canal has been described to be the most commonly affected part of the inner ear in humans, and flouncer provides a mouse model for genetic and developmental analysis of such defects. PMID- 15354000 TI - Early onset and rapid progression of dominant nonsyndromic DFNA36 hearing loss. AB - OBJECTIVE: To characterize the auditory and vestibular phenotype of autosomal dominant nonsyndromic DFNA36 hearing loss. STUDY DESIGN: Clinical evaluation of individuals with DFNA36 hearing loss linked to the D572N mutation of transmembrane channel-like gene 1 (TMC1). Medical history interviews, physical examinations, and pure-tone air conduction audiometry were performed in the field. Audiology and radiology reports were available and retrospectively reviewed for a subset of subjects. SETTING: Primary, secondary, and tertiary referral centers (retrospectively reviewed studies); subjects' homes (prospective clinical evaluations). PATIENTS: Thirteen affected members of a North American Caucasian family segregating DFNA36 hearing loss. MAIN OUTCOME MEASURES: Pure tone audiometric thresholds and their rates of progression. RESULTS: Subjects had bilateral, symmetric, sensorineural hearing loss with a postlingual onset in the first decade of life. High frequencies were initially affected, followed by rapid progression (5.9 dB/yr for the 0.5/1/2/4-kHz pure-tone average) to profound deafness across all frequencies by the second decade of life. Two individuals had excellent auditory-verbal communication after rehabilitation with cochlear implants placed over two decades after total deafening. CONCLUSIONS: DFNA36 has one of the earliest onsets and most rapid rates of progression among the autosomal dominant non-syndromic hearing loss phenotypes. These distinctive features should facilitate its clinical detection and the development of clinical molecular genetic diagnostic algorithms for dominant nonsyndromic hearing loss. PMID- 15354001 TI - Short-term versus long-term antibiotic prophylaxis in cochlear implant surgery. AB - OBJECTIVE: The objective of this study to evaluate antibiotic prophylaxis options for cochlear implant surgery. Does long-term antibiotic prophylaxis have any advantage over a single perioperative dose in preventing postoperative infection? STUDY DESIGN: We conducted a retrospective case review. PATIENTS: A total of 292 adult and pediatric patients who underwent cochlear implantation during a 15-year period (1988-2003) were reviewed. MAIN OUTCOME MEASURE: Minor and major postoperative wound infections in first 4 weeks. RESULTS: There were four major and eight minor complication among 292 patients (complication rate 4.1%). The infection rate was higher in patients who had either C incision (11.1%) or extended endaural incision (7.5%) and in patients with a preexisting medical condition. The infection rate was also higher in patients who had long-term antibiotics (5.6% and 13% in the 5-d and 7-d regimes) compared with short-term (single-dose) group. CONCLUSION: Long-term antibiotic prophylaxis did not have any advantage over single perioperative dose. Predisposing medical conditions and extensive surgical incisions were associated with a greater severity of infections and higher risk of wound complications. PMID- 15354002 TI - Late failure of cochlear implantation resulting from advanced cochlear otosclerosis: surgical and programming challenges. AB - OBJECTIVE: The objective of this study was to discuss cochlear implantation in the setting of severe cochlear otosclerosis and review programming challenges in a patient whose performance is deteriorating as a result of advancing disease. STUDY DESIGN: We conducted a case report and literature review. SETTING: Tertiary care medical center. PATIENTS, INTERVENTION, AND RESULTS: A case is presented of a 66-year-old man with otosclerosis who initially had good benefit from a cochlear implant but gradually lost benefit even with reimplantation. Imaging studies demonstrated severely distorted otic capsule anatomy from cochlear otosclerosis. CONCLUSIONS: Advancing cochlear otosclerosis can result in a severely thinned and distorted otic capsule. Although cochlear implantation is generally beneficial in cochlear otosclerosis, implantation in certain severe cases may be complicated as a result of difficulties with cerebrospinal fluid leak, programming challenges, and other potential hazards that can occur with a distorted anatomy. PMID- 15354003 TI - Labyrinth dysfunction 8 months after cochlear implantation: a case report. AB - OBJECTIVE: Investigate whether the cochleostomy is a possible port of entry for pneumolabyrinth and a resulting vertigo in patients provided with a cochlear implant. STUDY DESIGN: Retrospective case review. SETTING: Ludwig-Maximilians University of Munich, Hospital Grosshadern. PATIENT: 62-year-old patient who underwent implantation of a HiFocus II cochlear implant with positioner from Advanced Bionics (CLARION). Eight months postoperatively, the patient reported rotatory vertigo and right-side tinnitus after he had blown his nose harder than usual during an episode of rhinitis. INTERVENTIONS: Preoperative and postoperative testing of both the petrosal bone with a CT scan and of balance function. MAIN OUTCOME MEASURE: Air inclusion in the labyrinth. RESULTS: In contrast to the preoperative high resolution computed tomography (CT) scan, air inclusion was seen in the labyrinth during the episode of vertigo. At the same time, balance function tests with Frenzel glasses revealed both spontaneous and provoked horizontal nystagmus to the right side. At follow-up 8 weeks later, the level of vertigo had significantly decreased. Twelve months later, the control CT showed the cochlear implant positioned correctly and no visible air in the labyrinth. CONCLUSION: It is known that placement of the HiFocus II with Positioner from CLARION requires a relatively large cochleostomy of 1.5 mm. Moreover, in the connective tissue seal between the electrode and the positioner, the latter reaches into the tympanic cavity, and this is possibly the weak point. Further investigation will be needed to determine whether the large cochleostomy with the HiFocus II with positioner increases the predisposition to labyrinth dysfunction. PMID- 15354004 TI - Surgical complications and their management in a series of 300 consecutive pediatric cochlear implantations. AB - OBJECTIVE: To report the short- and long-term complications encountered in a large number of consecutive children undergoing implantation in a single center. The current study also describes the management and sequelae of each complication. STUDY DESIGN: Prospective study assessing the surgical findings and complications of deaf children undergoing implantation. SETTING: Pediatric tertiary referral center for cochlear implantation. PATIENTS: The present study includes 300 consecutive children undergoing implantation, with a mean age at implantation of 5.1 years, ranging from 1.3 to 16.9 years. Of these children, 196 (65%) had congenital deafness of unknown cause. The commonest known cause was meningitis (73 of 300 [24%]) followed by congenital cytomegalovirus infection (17 of 300 [6%]). Children have been followed up regularly after implantation, typically at yearly intervals after the first year. The mean duration of follow up at the time of the study was 4 years (range, 0.1-14 yr). RESULTS: There were no major perioperative (within 1 d after surgery) or major early postoperative (within 1 wk after surgery) complications. In the same periods, there were 19 and 15 minor complications, respectively. These complications (e.g., eardrum perforation, hematoma, flap swelling, wound infection, temporary facial weakness) settled with conservative treatment or minor intervention. With regard to the late surgical complications (>1 wk after surgery), there were 7 major (e.g., severe flap infection requiring explantation, cholesteatoma, persistent eardrum perforation) and 14 minor complications (e.g., mild flap infection, flap swelling, hematoma). A number of complications were encountered even 14 years after the original operation, and some of them needed repeated interventions, highlighting the importance of long-term follow-up. However, most of the complications occurred very close to the surgical procedure (<1 yr). CONCLUSION: An overall rate of 2.3% for major surgical complications and an overall rate of 16% for minor surgical complications suggest that cochlear implantation is a relatively safe surgical operation in experienced centers. Most surgical complications are minor and can be managed with conservative treatment or minor surgical intervention. However, meticulous attention to surgical detail, especially handling soft tissues and leaving the posterior canal wall intact, and long-term follow-up are of paramount importance in minimizing the incidence of surgical complications. PMID- 15354005 TI - Stimulation of the semicircular canals via the rotary chair as a means to test pharmacologic countermeasures for space motion sickness. AB - OBJECTIVE: Space motion sickness is currently treated pharmacologically with the empiric use of the H1 antihistamine promethazine, but use of this intervention is limited by the side effect of significant sedation. This creates a dilemma, as full cognition is particularly important during the same conditions likely to exacerbate the symptoms of space motion sickness. Using overstimulation of the semicircular canals with a rotary chair as a paradigm for space motion sickness, we evaluated four medications, commonly used for the treatment of terrestrial motion sickness and vertigo, for their efficacy in alleviating the simulated symptoms of space motion sickness. STUDY DESIGN: Randomized, prospective, double blind study. SETTING: Tertiary referral center. PATIENTS: Healthy male and female volunteers, 18 years of age or older, without history of neurologic or psychiatric disorders, and with no known allergies or any previous adverse reactions to the drugs used. INTERVENTIONS: Lorazepam 1 mg, meclizine 25 mg, promethazine 25 mg, scopolamine 0.4 mg, or placebo. MAIN OUTCOME MEASURE: The ability of each treatment to control the nausea and vomiting associated with our paradigm for space motion sickness was evaluated by measuring time of rotation pre- and posttreatment and time of symptom onset pre-and posttreatment. RESULTS: Only scopolamine effected a mean change in duration of rotation that reached statistical significance when compared with placebo (p <0.008), with a greater than 40% increase in rotation time. Results with promethazine were not statistically significant. CONCLUSION: Results showed a rank order of efficacy of scopolamine > promethazine > placebo > meclizine > lorazepam. Scopolamine significantly increased rotation time, but none of the treatments resulted in a significant delay to onset of symptoms. PMID- 15354006 TI - Prediction of fall risk reduction as measured by dynamic gait index in individuals with unilateral vestibular hypofunction. AB - OBJECTIVE: To determine the effect of vestibular rehabilitation on reduction of fall risk in individuals with unilateral vestibular hypofunction and to identify those factors that predict fall risk reduction. STUDY DESIGN: Retrospective chart review. SETTING: Tertiary referral center. PATIENTS: Forty-seven patients with unilateral vestibular hypofunction, aged 28 to 86 years, who were at risk for falls on initial assessment. INTERVENTION: All patients underwent vestibular rehabilitation including adaptation exercises, designed to improve gaze stability, and gait and balance exercises. MAIN OUTCOME MEASURES: Fall risk (Dynamic Gait Index), visual acuity during head movements (Dynamic Visual Acuity), and subjective complaints were measured initially, at 2-week intervals, and at completion of physical therapy. RESULTS: As a group, the patients had significantly reduced risk for falls (p <0.001) after rehabilitation. Time from onset of symptoms did not affect the efficacy of vestibular rehabilitation. Both older (> or = 65 yr) and younger (< 65 yr) adults showed significant reductions in fall risk with vestibular rehabilitation (p <0.001). However, a significantly greater proportion (Chi2= 0.016) of older adults remained at risk for falls at discharge compared with young adults (45% versus 11%). Initial Dynamic Gait Index and Dynamic Visual Acuity scores predicted fall risk reduction in patients with unilateral vestibular hypofunction. A model was developed using initial Dynamic Gait Index and Dynamic Visual Acuity scores to predict fall risk reduction. CONCLUSIONS: Vestibular rehabilitation is effective in significantly reducing fall risk in individuals with unilateral vestibular deficit. The model predicts fall risk reduction with good sensitivity (77%) and specificity (90%). PMID- 15354007 TI - Gamma knife radiosurgery for acoustic neuromas performed by a neurotologist: early experiences and outcomes. AB - OBJECTIVE: To assess early outcomes after Gamma knife radiosurgery of acoustic neuromas and other skull base tumors. BACKGROUND: Gamma knife radiosurgery is one of the available methods to treat acoustic neuromas, in addition to micro surgical resection. Neuro-otologists have long been associated with microsurgical resection of these tumors; however, the application of Gamma knife radiosurgery to the treatment of these tumors by neuro-otologists has not been previously described. SETTING: Acoustic Neuroma and Skull Base Surgery Program / Tertiary Referral Center. STUDY DESIGN/PATIENTS/INTERVENTION: Prospective clinical study of all patients treated by the senior author and our gamma knife team beginning in June 2000. MAIN OUTCOME MEASURES: Preoperative MRI, audiometry, vestibular testing and facial nerve electromyography were completed. At six-month intervals postoperatively, audiometry, caloric testing and MRI were performed to determine thresholds and speech discrimination ability, vestibular function, and the size of the tumor. RESULTS: From June 2000 until March 2004, 38 patients were treated, and these included 33 acoustic neuromas, two meningiomas, one glomus jugulare tumor, and two facial neuromas. Greater than 36 month follow-up was available in 7 patients, > 24 months in 24, > 12 months in 31, and > 6 months in 34 patients. Statistically significant reduction in tumor size was seen over time, and tumor control was achieved in all but two patients. Various patterns of changes in auditory function, both in threshold and speech discrimination were observed in either positive or negative directions. CONCLUSIONS: Preliminary experience with Gamma knife radiosurgery indicates that this treatment method represents another option for neuro-otologists to use in managing patients with skull base tumors. PMID- 15354008 TI - Extended middle fossa approach to the petroclival junction and anterior cerebellopontine angle. AB - OBJECTIVES: This article seeks to demonstrate the use of the extended middle cranial fossa approach in the treatment of tumors arising in the anterior cerebellopontine angle and petroclival region. STUDY DESIGN: We conducted a retrospective chart review. SETTING: Tertiary referral center. PATIENTS: : Ten year retrospective chart review of over 800 skull base surgical cases demonstrated 16 cases in which the senior author used the extended middle cranial fossa as the sole approach to access the posterior cranial fossa, petroclival junction, or the anterior cerebellopontine angle. There were five males and 11 females, 13 meningiomas, 2 trigeminal schwannomas, and 1 brainstem glioma. Presenting symptoms were dependent on extent of brainstem compression and involvement of surrounding cranial nerves. The symptoms are broken down as follows: hydrocephalus, one; balance disturbance, three; diplopia, five; trigeminal neuralgia, two; hemifacial numbness, one; seizures, one; expressive aphasia, one; and hearing loss, two. RESULTS: Of the 16 patients in this study, one patient needed postoperative care in a skilled nursing facility. Postoperative facial nerve weakness was not experienced in any patient. One patient developed a transient cerebrospinal fluid leak that resolved spontaneously. One patient developed a pseudomeningocele secondary to postoperative hydrocephalus. This was corrected with wound exploration and placement of a ventricular peritoneal shunt. Hearing was not maintained in one patient. Two patients developed new fourth nerve paresis and two patients developed new sixth nerve palsies. There were no postoperative infections and no deaths. CONCLUSIONS: The extended middle cranial fossa approach provides excellent access and exposure to tumors in the anterior cerebellopontine angle and petroclival junction. The approach allows more direct access to the area anterior to the internal auditory canal. The key to the approach is adequate bone removal of the petrous apex to provide exposure down to the inferior petrosal sinus and anteriorly to Meckel's cave and the petroclival junction. Extradural elevation of the temporal lobe with suitable brain relaxation minimizes postoperative complications. PMID- 15354009 TI - Opening cerebrospinal fluid pressure guides the management of cerebrospinal fluid leakage after acoustic neuroma surgery. AB - OBJECTIVE: To assess the value of cerebrospinal fluid pressure as a decisional factor for immediate surgical revision in cerebrospinal fluid leakage after acoustic neuroma removal. STUDY DESIGN: Prospective study. SETTING: Tertiary referral center. PATIENTS: Between 1998 and 2001, 220 patients were operated on for acoustic neuroma by different transpetrosal approaches. Among 24 patients (12%) presenting postoperative cerebrospinal fluid leakage, those with meningitis or with hydrocephalus were excluded. Fifteen patients were included in this study. METHODS: Each patient had initial conservative treatment with serial depletive lumbar punctures and cerebrospinal fluid pressure measurements associated with oral acetazolamide. Surgical revision was decided on in case of persistent cerebrospinal fluid leakage. RESULTS: In eight patients with high cerebrospinal fluid pressure (18+/-1.4 cm H2O; range, 14-28 cm H2O), cerebrospinal fluid leak disappeared in 3 days after conservative treatment. Seven other patients required surgical revision for persistent cerebrospinal fluid leakage. Revision surgery was efficient in six patients with low cerebrospinal fluid pressure (8+/-1.3 cm H2O, range, 3-12 cm H2O). In the remaining patient with high cerebrospinal fluid pressure (18 cm H2O), cerebrospinal fluid leakage continued despite surgical revision, requiring lumboperitoneal shunting. CONCLUSION: The cerebrospinal fluid pressure value may be used as a decisional indicator for cerebrospinal fluid leakage treatment after acoustic neuroma surgery. Low cerebrospinal fluid pressure leakage would imply a revision surgery procedure without delay, whereas high cerebrospinal fluid pressure leakage would imply conservative treatment. PMID- 15354010 TI - Differential grading of endolymphatic sac tumor extension by virtue of von Hippel Lindau disease status. AB - OBJECTIVE: Endolymphatic sac tumors are aggressive papillary tumors of the temporal bone frequently associated with von Hippel-Lindau disease. The goal of this study was to use a newly devised classification system as a means to analyze differences between endolymphatic sac tumor extension in von Hippel-Lindau disease and non-von Hippel-Lindau disease patients. METHODS: Previously reported cases of endolymphatic sac tumor and two new cases were retrospectively reviewed and assigned to a new classification system consisting of four grades based on tumor extent and location. RESULTS: Mean age of 103 patients without von Hippel Lindau disease was 52.5 years, whereas in 46 patients with VHL the mean age was 31.3 years. Patients with von Hippel-Lindau disease were more likely to be female (female-to male ratio of 2:1 for von Hippel-Lindau disease patients versus 1:1 for non-von Hippel-Lindau disease patients). Symptoms consisted of hearing loss (100% [mean duration, 10 yr] for VHL patients versus 97% [mean duration, 7.8 yr] for non-von Hippel-Lindau disease patients), facial weakness (38% versus 49%), and tinnitus or vertigo (41% versus 60%). Bilateral tumors were common in von Hippel-Lindau disease patients (28% versus 1%). Tumors in von Hippel-Lindau disease patients were significantly more likely to be lower grade than tumors in non-von Hippel-Lindau disease patients (Grade I, 40% versus 25%; Grade II, 50% versus 58%; Grade III, 8% versus 14%; and Grade IV, 2% versus 4%; p < 0.05). Before 1988, there were relatively fewer Grade I (15% versus 33%) and relatively more Grade II (69% versus 47%) endolymphatic sac tumors in non-von Hippel-Lindau disease patients than after 1988. CONCLUSIONS: Increased usefulness of intracranial imaging since 1988 has led to the diagnosis of sporadic endolymphatic sac tumors with lower grades. Surveillance imaging in von Hippel Lindau disease may account for the greater proportion of endolymphatic sac tumors diagnosed with lower grades. Endolymphatic sac tumors associated with a diagnosis of von Hippel-Lindau disease appear to affect a younger population of patients than non-von Hippel-Lindau disease cases and occur in women twice as often as in men when associated with von Hippel-Lindau disease. In addition, tumors are more frequently bilateral and less advanced in the von Hippel-Lindau disease patient as opposed to the non-von Hippel-Lindau disease patient. PMID- 15354011 TI - Endolymphatic sac tumor in a 4-year-old boy. AB - INTRODUCTION: Endolymphatic sac tumors (ELST) are rare, low-grade, locally aggressive papillary neoplasms. We present a case of a 4-year-old boy with an ELST, the youngest described in the literature. CASE: A boy presented with a right-sided serous otitis media and sudden-onset right facial nerve palsy. An audiogram revealed right-sided profound sensorineural hearing loss. Radiographic imaging demonstrated a 3-cm expansile lytic lesion along the posterior face of the petrous bone. INTERVENTION/RESULTS: The patient initially underwent a right transmastoid-infralabyrinthine biopsy. Pathologic examination revealed a papillary lesion suspicious for an ELST. Subsequently, a transtemporal transcochlear approach with intra-and extradural resection of the tumor was performed. The facial nerve was dissected and transposed anteriorly and preserved. Histopathologic and immunohistochemical studies confirmed the ELST. At his 6-month follow up, there is no evidence of recurrence and the facial nerve function has returned to Grade II palsy. CONCLUSION: ELST are rare tumors of the temporal bone. This is the youngest case of ELST reported. Presentation, evaluation, and management of ELST is discussed. PMID- 15354012 TI - Clinical and radiologic sequelae of the middle fossa approach to the internal auditory canal. AB - OBJECTIVE: This retrospective study aimed to characterize the postoperative nonneurotologic complications and radiologic findings after the middle fossa approach to the internal auditory canal (IAC). A grading system for computed tomography (CT) changes was developed. STUDY DESIGN: Sixty-two patients operated between 2000 and 2003 had CT scans of the head on the day after surgery. Their charts and films were reviewed. SETTING: R15 DK 54070-01A1: Tertiary referral center. RESULTS: Nine patients had complications, including seven with cerebrospinal fluid leakage, one with infection, and one with visual disturbances. Only three of these patients had CT changes. Alternatively, 14 patients had CT changes according to our grading system for edema, pneumatocephalus, and hematoma. Three of these patients had complications. The overall complication rate was low (15%), and there were no cases of serious damage to the temporal lobe. CONCLUSION: The middle fossa approach provides safe access to the IAC. With the use of flexible retractors, serious damage to the temporal lobe can be avoided. The CT scan on the first day after surgery is useful for detecting dangerous hematomas but should be correlated to the patient's condition because of non-specific radiologic changes and the potential for complications to be present even in the absence of CT findings. PMID- 15354013 TI - Correlation of growth factor receptor expression with clinical growth in vestibular schwannomas. AB - OBJECTIVE: To examine the relationship between growth rate of vestibular schwannomas and the expression of various growth factor receptors. STUDY DESIGN: Retrospective case review of clinical growth rate in conjunction with a histopathologic and immunohistochemical reexamination of archival specimens. SETTING: A tertiary referral neurotologic center. PATIENTS: Three groups: a historical group to act as controls, consisting of 30 patients with sporadic vestibular schwannomas removed before the unit adopted an initial interval scan policy; a group of 14 patients with sporadic vestibular schwannomas who had undergone an initial interval scan policy, showed radiologic evidence of growth, and therefore had their schwannoma removed; and a group of 16 schwannomas removed from 11 neurofibromatosis Type 2 patients. MAIN OUTCOME MEASURES: A comparison between the three clinical groups using immunohistochemical studies to determine the level of expression of the proliferation factor Ki-67, c-erbB-2, and c-erbB-3 receptors and fibroblastic growth factor receptors 1 and 4. RESULTS: The level of expression of the proliferation factor Ki-67 was very low and similar in all three groups. C-erbB-2 and c-erbB-3 receptors were not expressed in any of the groups. fibroblastic growth factor receptor 4 expression was not significantly different, but there was a variation in the expression of fibroblastic growth factor receptor 1 between the three groups that correlated well with the differing incidence of growth in the groups. The increase in expression of fibroblastic growth factor receptor 1 in the neurofibromatosis Type 2 group was not statistically significant, but the increase in expression of fibroblastic growth factor receptor 1 in the growing sporadic group was statistically significant when compared with the historical controls. The level of fibroblastic growth factor receptor 1 expression correlates significantly with the rate of growth as measured on interval magnetic resonance imaging. CONCLUSIONS: Overexpression of fibroblastic growth factor receptor 1 has a positive correlation with the incidence and the rate of growth of sporadic vestibular schwannomas. PMID- 15354014 TI - Management of jugular paragangliomas: the Gruppo Otologico experience. AB - OBJECTIVE: The objective of this study was to review the outcome of surgical management in patients of jugular paragangliomas. STUDY DESIGN: We conducted a retrospective case review. SETTING: Tertiary care otology and skull base center. MATERIALS AND METHODS: Fifty-five patients with the diagnosis of a jugular paraganglioma (Fisch Class C and D Glomus Jugulare) were managed over a period of 15 years. All patients with adequate follow up and complete records (53 cases) were reviewed with emphasis on the results of surgical management and the factors influencing them. INTERVENTION: All 53 patients were managed with a view to surgically extirpate the tumor. The primary approach was the infratemporal fossa approach-Type A used in the majority of the patients. In eight cases, the procedure was staged owing to the presence of large intracranial extension. Three patients required additional procedures to ameliorate the after-effects of lower cranial nerve resection. RESULTS: Gross total tumor removal was achieved in 49 patients. There were five cases of recurrence. Coupled with the residual tumors in five patients, the surgical control achieved was 83%. There was no perioperative mortality. There were two cases of postoperative cerebrospinal fluid leak, both of which required surgical exploration and closure. The facial nerve was resected in seven patients. The overall preservation rate of clinically uninvolved lower cranial nerves was 75%. CONCLUSIONS: The low level of complications along with a high surgical control achieved makes surgery the primary mode of treatment in the vast majority of these tumors, regardless of the size and location. PMID- 15354015 TI - Delayed facial paralysis after vestibular schwannoma surgery: role of herpes viruses reactivation--our experience in eight cases. AB - OBJECTIVES: The objectives of this study were to study the role of herpes virus reactivation in the onset of delayed facial paralysis (DFP) occurring more than 72 hours after vestibular schwannoma (VS) surgery and to advocate specific medical management. STUDY DESIGN: We conducted a retrospective case review. SETTING: University-based, tertiary care center. PATIENTS: Eight patients managed for DFP in a series of 348 patients operated for a VS. INTERVENTIONS: Patients were evaluated and graded according to the House and Brackmann grading system and followed up for 1 year. A serologic search for specific antiherpes simplex viruses type 1 and 2 (HSV-2) and varicella zoster virus (VZV) antibodies at the onset of DFP and 2 weeks later was possible in three cases. Seven of the eight patients were given intravenous acyclovir (30 mg/kg/ for 5 days) and methylprednisolone (2 mg.kg/ for 7 days). RESULTS: Mean delay of DFP onset was 8.75 days. All treated patients had a House and Brackmann Grade 1 recovery: mean time to recovery was 40.4 days. The last one had only a Grade 3 recovery because he could not be treated because of postoperative transient psychiatric problems. Serologic testing in those patients in whom it could be done revealed either a high level of anti HSV or VZV antibodies at the time of onset or a dramatic increase in anti-HSV or anti-VZV antibodies between the two samples, strongly suggesting an HSV or VZV reactivation. CONCLUSION: HSV or VZV reactivation might be responsible for most cases of DFPs, thus suggesting the usefulness of immediate steroid and acyclovir administration to obtain total recovery. The viral reactivation mechanism is comparable to that already suspected in DFP occurring with the same delay in middle ear surgical procedures. PMID- 15354016 TI - Vestibular schwannoma growth rates in neurofibromatosis type 2 natural history consortium subjects. AB - OBJECTIVE: To determine the amount of growth in vestibular schwannomas in Neurofibromatosis type 2 (NF2) patients from diagnosis through short-term (up to 2 yr) and long-term (up to 4 yr) follow-up. STUDY DESIGN: Retrospective magnetic resonance imaging (MRI) films were obtained on subjects enrolled in the NF2 Natural History study and examined for changes in vestibular schwannoma size over time. SETTING: Data were collected from nine foreign and domestic NF2 centers, including hospital-based, academic, and tertiary care centers. SUBJECTS: NF2 patients with MRI data and at least one follow-up examination within 9 months to 2 years of diagnosis were included; n=56 patients with 84 lesions for evaluation of growth. INTERVENTION: Routine, clinically obtained, magnetic resonance images were digitized and measured using image management software. Short-term follow-up was defined as up to 2 years (n=84 lesions), and long-term follow-up was defined as 3 to 4 years (n=29 lesions). OUTCOME MEASURES: Vestibular schwannoma size was assessed using anterior-posterior, medial-lateral, and greatest diameter linear measurements. RESULTS: Vestibular schwannomas increased in size (at least 5 mm) in 8% of the vestibular schwannomas across short-term follow-up. At long-term follow-up, 13% of the tumors had increased in size. On average, schwannomas increased in greatest diameter 1.3 mm per year across short-term follow-up. CONCLUSION: Slightly greater than 1 in 10 diagnosed NF2-related vestibular schwannomas increased in size by at least 5 mm by 4 years of follow-up, if still untreated at that time. PMID- 15354017 TI - Intraoperative monitoring of cochlear function using distortion product otoacoustic emissions (DPOAEs) in patients with cerebellopontine angle tumors. AB - HYPOTHESIS: Intraoperative monitoring by distortion-product otoacoustic emissions reflects the cochlear function changes in the real-time domain during removal of cerebellopontine angle tumors. BACKGROUND: Cerebellopontine angle tumor surgery is associated with a significant risk of damaging internal auditory canal contents. Although monitoring facial nerve function intraoperatively has already been effectively developed, such efficacious monitoring of auditory function remains to be established. The aim of this study was to investigate the utility of distortion-product otoacoustic emissions for intraoperative monitoring of the cochlear function in humans during removal of cerebellopontine angle tumors. METHODS: Continuous intraoperative monitoring of distortion-product otoacoustic emissions was performed in 20 of 62 patients undergoing surgical removal of cerebellopontine angle tumors. All of these 20 patients, who underwent the retrosigmoid approach, had distortion-product otoacoustic emissions present preoperatively. Depending on the amplitude and frequency band at which distortion product otoacoustic emissions were present, monitoring was carried out at 2.0 to 6.0 kHz with primary stimulus tone amplitudes of 60 to 70 dB sound pressure level. RESULTS: In patients operated on for cerebellopontine angle tumors, various patterns of distortion-product otoacoustic emission amplitude reductions and recoveries were observed. Distortion-product otoacoustic emissions recorded from the basal part of the cochlea (i.e., high frequencies) changed earlier and more profoundly than those from the middle and apical sections (i.e., lower frequencies). In some cases, cochlear function was affected irreversibly as reflected by loss of distortion-product otoacoustic emissions. Microcoagulation of small vessels, tumor debulking, and compression or stretch of the internal auditory canal contents were found to be procedures affecting distortion-product otoacoustic emissions. The status of distortion-product otoacoustic emissions at the conclusion of tumor dissection correlated with postoperative hearing levels. CONCLUSION: Distortion-product otoacoustic emissions were used to monitor in the real-time domain auditory function during cerebellopontine angle tumor removal operations. The status of distortion-product otoacoustic emissions at the conclusion of the operations was related to postoperative hearing. PMID- 15354018 TI - A comparison of direct eighth nerve monitoring and auditory brainstem response in hearing preservation surgery for vestibular schwannoma. AB - OBJECTIVE: The objective of this study was to compare the effectiveness of direct eighth nerve monitoring (DENM) and auditory brainstem response (ABR) in facilitating hearing preservation during vestibular schwannoma resection. STUDY DESIGN: This was a retrospective study. SETTING: : Tertiary referral center. METHODS: We conducted a retrospective clinical study of the use of ABR and DENM during vestibular schwannoma removal. Tumors were removed through a retrosigmoid craniotomy. The rate of hearing preservation between the two monitoring modalities was compared. The additional outcome measures of facial nerve function and cerebral spinal fluid leak rate were also evaluated. RESULTS: Hearing preservation was attempted in 77 patients with vestibular schwannomas. Tumor sizes ranged from 0.5 cm to 2.5 cm. Hearing was preserved in 71% of patients with tumors 1 cm or less and in 32% of patients with tumors between 1 and 2.5 cm when direct eighth nerve monitoring was used. Hearing preservation rates with ABR for tumors 1 cm or less were 41% and 10% in patients with tumors between 1 and 2.5 cm (p=0.03) Facial nerve preservations rates were 94% (House-Brackmann 1-2) for tumors less than 2 cm. CONCLUSIONS: DENM provides significantly higher rates of hearing preservation during vestibular schwannoma resection when compared with ABR. PMID- 15354019 TI - Hearing loss in Wegener's granulomatosis. AB - OBJECTIVE: To describe the frequency, type, and clinical course of hearing loss in Wegener's granulomatosis and assess hearing loss as an indicator of disease activity. STUDY DESIGN, SETTING, AND PATIENTS: Retrospective cohort review of all patients with Wegener's granulomatosis seen in 1 year at an academic medical center. MAIN OUTCOME MEASURES: Hearing loss documented by pure-tone audiogram. RESULTS: Thirty-six patients were included in the analysis: 20 men and 16 women, with a mean age of 55.5 years (range, 22-87 yr); 30 (83%) were antineutrophil cytoplasmic autoantibodies-positive, and the mean disease duration was 47 months (range, 2-196 mo). Twenty patients (56%) had documented hearing loss: there were 17 (47%) cases of sensorineural hearing loss and 12 (33%) cases of conductive hearing loss. Seven of 12 cases of conductive hearing loss improved with immunosuppressive treatment of Wegener's granulomatosis, 2 worsened, and 3 remained stable. Of 17 cases of sensorineural hearing loss, 3 improved, 4 worsened, and 10 remained stable. Seven patients had hearing loss requiring amplification. Five of 35 (14%) patients had established hearing loss months to years before diagnosis of Wegener's granulomatosis. Hearing loss occurred both on initial presentation and with disease relapse. The rates of conductive hearing loss (38%) and sensorineural hearing loss (31%) were also high in the subset of patients 65 years of age or younger and without history of noise exposure. CONCLUSIONS: Both sensorineural hearing loss and conductive hearing loss are common in Wegener's granulomatosis, may result in significant morbidity, and may precede the diagnosis of Wegener's granulomatosis by years. Both types of hearing loss in patients with Wegener's granulomatosis may be used as an indicator of disease. These data suggest that it may be appropriate to obtain screening audiograms in all patients with newly diagnosed or relapsing Wegener's granulomatosis. PMID- 15354020 TI - The VlsE (IR6) peptide ELISA in the serodiagnosis of lyme facial paralysis. AB - OBJECTIVE: Facial paralysis is a manifestation of early disseminated Lyme neuroborreliosis. In the current study, we compared the immunoglobulin G (IgG) VlsE (sixth invariant region) peptide enzyme-linked immunosorbent assay (ELISA) with the current two-tier approach of sonicate ELISA and Western blot in the serodiagnosis of Lyme facial paralysis. STUDY DESIGN: Retrospective. SETTING: Tertiary referral center. PATIENTS: Serum samples from 47 Lyme patients with facial paralysis and 86 control subjects were analyzed for IgG antibodies to VlsE peptide of Borrelia burgdorferi and for immunoglobulin M (IgM) and IgG antibodies to sonicate antigens of B. burgdorferi using the two-tier approach. INTERVENTION: Diagnostic. MAIN OUTCOME MEASURE: Serum IgG antibody responses to VlsE (IR6) peptide. RESULTS: All 47 (100%) patients with facial paralysis and 4 (5%) of 86 controls had positive antibody responses to the VlsE peptide. In the two-tier test, 41 (87%) patients had positive IgM, 31 (66%) had positive IgG, and all 47 patients had positive IgM or IgG responses. Of the 86 control subjects, 2 (2%) had positive results with the two-tier test. Thus, the sensitivities of the VlsE and the two-tier tests were 100%; the specificity of the VlsE ELISA was 95% and the specificity of the two-tier test was 98%. CONCLUSIONS: The VlsE peptide ELISA showed a high sensitivity and specificity in the serological diagnosis of Lyme facial paralysis, similar to the two-tier test. The principal advantage of the VlsE peptide ELISA is that it requires only one test rather than four tests. However, the specificity of the VlsE test may not be as high as that of the two tier test. PMID- 15354021 TI - Proteomics: a primer for otologists. AB - OBJECTIVE: On July 9, 2003, the National Institutes of Health (NIH) released a new program announcement entitled "Proteomics in Auditory and Developmental Disease Processes." This initiative makes it clear that proteomic analysis in otology is a multi-year research priority for the NIH. The goal of this article is to describe the mechanics of modern proteomic techniques and review their applications in otology to date. DATA SOURCES: General articles from the proteomic literature were used to construct a review of modern proteomic techniques. For literature on proteomics in otology, MEDLINE and CRISP databases were searched by various topics in otology and cross-referenced with principle proteomic technologies. STUDY SELECTION: The criterion for selection was any study in otology that employs proteomic technology. CONCLUSIONS: Incredible progress has been made in proteomic technology. However, modern proteomic techniques are currently underutilized in otologic research. The NIH proteomics initiative referenced above, in combination with an understanding of the basic tools of modern proteomic science, should help motivate otologists to discover innovative ways to apply modern proteomic techniques to specific problems in otology. PMID- 15354022 TI - Combination of aberrant internal carotid artery and persistent stapedial artery. PMID- 15354023 TI - Transverse fracture of the temporal bone. PMID- 15354024 TI - The human gene map for performance and health-related fitness phenotypes: the 2003 update. AB - This review presents the 2003 update of the human gene map for physical performance and health-related fitness phenotypes. It is based on peer-reviewed papers published by the end of 2003 and includes association studies with candidate genes, genome-wide scans with polymorphic markers, and single-gene defects causing exercise intolerance to variable degrees. The genes and markers with evidence of association or linkage with a performance or fitness phenotype in sedentary or active people, in adaptation to acute exercise, or for training induced changes are positioned on the genetic map of all autosomes and the X chromosome. Negative studies are reviewed but a gene or locus must be supported by at least one positive study before being inserted on the map. By the end of 2000, 29 loci were depicted on the first edition of the map. In contrast, the 2003 human gene map for physical performance and health-related phenotypes includes 109 autosomal gene entries and QTL, plus two on the X chromosome. Moreover, there are 15 mitochondrial genes in which sequence variants have been shown to influence relevant fitness and performance phenotypes. PMID- 15354025 TI - Similar histopathological picture in males with Achilles and patellar tendinopathy. AB - PURPOSE: To ascertain whether there are differences in the histopathological appearance of tendinopathic Achilles and patellar tendons. METHODS: In males, we studied biopsies from tendinopathic Achilles (N = 28; average age 34.1 yr) and patellar tendons (N = 28; average age 32.1), Achilles tendons (N = 21; average age 61.8 yr) from deceased patients with no known tendon pathology, and patellar tendons (N = 15; average age 28.3) from patients undergoing anterior cruciate ligament reconstruction. Hematoxylineosin stained slides were interpreted using a semiquantitative grading scale (0: normal to 3: maximally abnormal) for fiber structure, fiber arrangement, rounding of the nuclei, regional variations in cellularity, increased vascularity, decreased collagen stainability, and hyalinization. All slides were assessed blindly twice, the agreement between two readings ranging from 0.170 to 0.750 (kappa statistics). RESULTS: The highest mean score of tendinopathic Achilles tendons was not significantly different from that of tendinopathic patellar tendons (11.6 +/- 5 and 10.4 +/- 3, respectively). The ability to differentiate between an Achilles tendon and a patellar tendon was low. CONCLUSIONS: Tendinopathic Achilles and patellar tendons show a similar histological picture. It was not possible to identify whether a specimen had been harvested from an Achilles or a patellar tendon on the basis of histological examination. The general pattern of degeneration was common to both tendinopathic Achilles and patellar tendons. A common, as yet unidentified, etiopathological mechanism may have acted on both these tendon populations. PMID- 15354026 TI - Exercise-induced apoptosis of lymphocytes depends on training status. AB - PURPOSE: To investigate the effect of training status on lymphocyte apoptosis as well as the expression of cell death receptors and ligands after a marathon run, and to compare these data with the alterations after treadmill exercise tests. METHODS: Sixteen volunteers successfully finished the 2002 Munster marathon. Venous blood samples were drawn before and 0, 3, and 24 h after the race. After cell isolation, cell-based apoptosis markers annexin V, Fas receptor, and Fas ligand were measured by flow cytometry. The same parameters were investigated in a group of 10 subjects before, and 0 and 1 h after both an exhaustive (ExT) and a low-intensity (LoT) treadmill test. RESULTS: The percentage of apoptotic cells after the marathon changed in a biphasic manner. An early increase 3 h after the run was followed by a significant decrease 1 d later. Interestingly, the increase in apoptotic cells was not observed in highly trained athletes, whereas it was significantly more pronounced in badly trained athletes. ExT induced a lymphocyte apoptosis similar to the marathon, whereas no change in apoptosis was observed after the LoT. Both Fas receptor and ligand were increased after the marathon with different kinetics. Whereas the Fas receptor peaked at 1 h, Fas ligand was increased 3 h after the run. After the treadmill tests Fas receptor expression was enhanced in both groups, whereas Fas ligand increased only after the ExT. CONCLUSIONS: Endurance exercise like a marathon is able to induce apoptosis in lymphocytes. Thereby, apoptosis sensitivity seems to be related to training status in an inverse relationship. The increased expression levels of death receptors and ligands might indicate the high apoptosis inducing potential of this type of exercise. PMID- 15354027 TI - Physical activity levels among breast cancer survivors. AB - INTRODUCTION: Obesity and weight gain are negative prognostic factors for breast cancer survival. Physical activity (PA) prevents weight gain and may decrease obesity. Little information exists on PA levels among cancer survivors. We assessed PA, including the proportion of breast cancer survivors engaging in recommended levels, by categories of adiposity, age, disease stage, and ethnicity in 806 women with stage 0-IIIA breast cancer participating in the Health, Eating, Activity, and Lifestyle Study. METHODS: Black, non-Hispanic white, and Hispanic breast cancer survivors were recruited into the study through Surveillance Epidemiology End Results registries in New Mexico, Western Washington, and Los Angeles County, CA. Types of sports and household activities and their frequency and duration within the third yr after diagnosis were assessed during an in person interview. RESULTS: Thirty-two percent of breast cancer survivors participated in recommended levels of PA defined as 150 min x wk(-1) of moderate- to vigorous-intensity sports/recreational PA. When moderate-intensity household and gardening activities were included in the definition, 73% met the recommended level of PA. Fewer obese breast cancer survivors met the recommendation than overweight and lean breast cancer survivors (P < 0.05). Fewer black breast cancer survivors met the recommendation compared with non-Hispanic white and Hispanic breast cancer survivors (P < 0.05). CONCLUSIONS: Most of the breast cancer survivors were not meeting the PA recommendations proposed for the general adult population. Efforts to encourage and facilitate PA among these women would be an important tool to decrease obesity, prevent postdiagnosis weight gain, and improve breast cancer prognosis. PMID- 15354029 TI - Comparative response of EPO and soluble transferrin receptor at high altitude. AB - PURPOSE: Soluble transferrin receptor (sTfR) classically raises with increased erythropoiesis, along with the rise in erythropoietin (EPO). However, the specific effect of altitude-induced erythropoiesis on sTfR remains poorly documented. This study investigated the response of sTfR during high-altitude exposure in human and verified that sTfR was related to EPO response in this case. METHODS: EPO, sTfR, red cell volume (RCV), ferritin, and iron intake were measured during: 1) experiment A (N = 8, 31 d at 5000-8848 m), at sea level (SL), and at the simulated altitude of 5000, 6000, 7000, and 8000 m; and 2) during experiment B (N = 10, 7 d at 4350 m), at SL, after 3, 5, and 7 d at 4350 m and 1 2 d after return to SL (RSL). RESULTS: In experiment A, progressive decompression from SL to 8000 m induced a large parallel rise in EPO (33.8-fold) and sTfR (5.9 fold), whereas ferritin was dramatically decreased and iron intake reduced. RCV was increased after 31 d of decompression. In experiment B, EPO peaked at day 3 at 4350 m, then declined later at altitude and returned to baseline values at RSL, whereas sTfR progressively rose at altitude (+86%) and remained elevated during RSL (+64%). Ferritin progressively declined at 4350 m, whereas iron intake was unchanged. RCV was not enhanced after exposure to 4350 m. CONCLUSION: In summary, sTfR mirrors EPO response for a given level of altitude hypoxia but differs from EPO response during transitory phases, such as early acclimatization or reoxygenation. Analysis of sTfR may therefore account for altitude-related erythropoiesis, at a time when EPO is blunted. PMID- 15354030 TI - Effects of sequential bouts of resistance exercise on androgen receptor expression. AB - PURPOSE: Increased serum testosterone (TST) occurs in response to resistance exercise and is associated with increased muscle mass. However, the effects of elevated TST and sequential resistance exercise bouts on androgen receptor (AR) expression in humans are not well known. This study examined three sequential bouts of heavy-resistance exercise on serum total TST, sex hormone-binding globulin (SHBG) and free androgen index (FAI), skeletal muscle AR mRNA and protein expression, and myofibrillar protein content. METHODS: Eighteen untrained males were randomly assigned to either a resistance-training [RST (N = 9)] or control group [CON (N = 9)]. RST performed three lower-body resistance exercise bouts, each separated by 48 h. At each exercise bout, RST performed three sets of 8-10 repetitions at 75-80% one-repetition maximum using the squat, leg press, and leg extension exercises, respectively, whereas CON performed no resistance exercise. Muscle biopsies were obtained immediately before the first exercise bout and 48 h after each of the three bouts, whereas blood samples were obtained immediately before, immediately after, and 30 min after each bout. Data were analyzed with two-way ANOVA and bivariate correlations. RESULTS: Serum TST and FAI were significantly increased after each exercise bout (P < 0.05); however, there were no significant changes for SHBG. AR mRNA and protein were significantly increased (P < 0.05) after the second and third exercise bouts, respectively, and were significantly correlated to TST and FAI (P < 0.05). Myofibrillar protein increased after the third bout (P < 0.05). CONCLUSIONS: Three sequential bouts of heavy resistance exercise increases serum TST and are effective at up-regulating AR mRNA and protein expression that appears to correspond to subsequent increases in myofibrillar protein. PMID- 15354031 TI - Left ventricular function in endurance-trained children by tissue Doppler imaging. AB - In children and adults, endurance training increases resting stroke volume, mainly as a result of an increase in left ventricular (LV) filling. PURPOSE: To evaluate whether the LV morphologic and functional alterations responsible for this increase in cardiac filling are similar in children and young adults. METHODS: Standard echocardiography (LV morphology and function) and tissue Doppler imaging (LV relaxation properties) were assessed in 10 adult cyclists, 13 age-matched sedentary controls, 12 boy cyclists, and 11 untrained boys. RESULTS: In our endurance-trained adults, LV morphological adaptations included increase in LV internal diameters, wall thickness, and mass. However, effects associated with training on LV morphology were different in children because no true cardiac hypertrophy was observed in our child cyclists compared with age-matched nonactive boys. Effects related training on LV systolic and diastolic function assessed by TDI were similar in boys and men. The LV diastolic function was improved in trained subjects (i.e., increased transmitral early to late filling velocities) as a result of an increase in LV relaxation properties. However, LV filling pressures, estimated from TDI, were similar in trained individuals compared with age-matched controls. CONCLUSION: In both children and adults, an increase in LV relaxation properties and normal LV filling pressures in endurance trained subjects might be taken as additional indicators for a physiologic or "normal" hypertrophy. However, further investigations are needed to evaluate whether the specific LV morphological adaptation observed in trained-children reflects a blunted trained-induced cardiac hypertrophy before puberty. PMID- 15354032 TI - Prevalence of exercise-induced arterial hypoxemia in healthy women. AB - PURPOSE: Exercise-induced arterial hypoxemia (EIAH) is reported to occur in approximately 50% of highly trained male endurance athletes. Few studies have examined EIAH in women and the prevalence remains unclear. It has been reported that some female subjects who develop EIAH possess maximal oxygen consumption (VO2max) values that are within 15% of their predicted value. This is unique to women, where EIAH has generally been reported in men who have a high VO2max. The primary objective of this investigation was to determine the prevalence of EIAH in a large female population with a wide range of VO2max values. It was hypothesized that EIAH would occur with a greater prevalence and at relatively lower predicted VO2max than that previously reported in males. METHODS: Young women (N = 52; 26.5 +/- 4.9 yr) performed a cycle test to exhaustion to determine VO2max, and oxyhemoglobin saturation (SaO2) was monitored via pulse oximetry. All subjects were tested during the early follicular phase of their menstrual cycle. A >/= 4% drop in SaO2 represented EIAH. RESULTS: Values for VO2max were variable (VO2max range: 28.0-61.3 mL x kg(-1) x min(-1)). EIAH was present in 67% of the women with N = 19 displaying mild EIAH (92-94%SaO2) and N = 16 displaying moderate EIAH (87-91%SaO2). CONCLUSION: It appears that the prevalence of EIAH in women is slightly greater than the 50% prevalence value that is typically reported for highly fit men. PMID- 15354033 TI - The impact of prolonged exercise in a cold environment upon cardiac function. AB - PURPOSE: The purpose of the present study was to examine the impact of cold exposure coupled with prolonged exercise upon postexercise left ventricular (LV) function and markers of myocardial damage. METHODS: colon; Eight highly trained male athletes (mean +/- SD; age: 28.2 +/- 8.8 yr; height: 1.78 +/- 0.07 m; body mass: 74.9 +/- 7.6 kg; VO2max: 65.6 +/- 7.0 mL x kg(-1) x min(-1)) performed two 100-mile cycle trials, the first in an ambient temperature of 0 degrees C, the second in an ambient temperature of 19 degrees C. Echocardiographic assessment was completed and blood samples drawn before, immediately postexercise, and 24-h postexercise. Left ventricular systolic (stroke volume [SV], ejection fraction [EF], and systolic blood pressure/end systolic volume ratio [SBP/ESV]) and diastolic (early [E] to late [A] filling ratio [E:A]) parameters were calculated. Serum was analyzed for creatine kinase isoenzyme MB (CK-MBmass) and cardiac troponin T (cTnT). cTnT was analyzed descriptively whereas other variables were assessed using two-way repeated-measures ANOVA. RESULTS: No significant change was observed in systolic function across time or between trials. A significant difference between trials was observed in E:A immediately after exercise (1.4 +/- 0.4 [19 degrees C] vs 1.8 +/- 0.3 [0 degrees C]) (P < 0.05). CK-MBmass was significantly elevated immediately after exercise in both trials (P < 0.05). Positive cTnT concentrations were observed in two subjects immediately after the 19 degrees C trial (0.012 microg x L(-1) and 0.034 microg x L(-1)). CONCLUSIONS: Cycling 100 miles in an ambient temperature of 19 degrees C is associated with an acute change in diastolic filling that is not observed after prolonged exercise at 0 degrees C. Prolonged exercise is associated with minimal cardiac damage in some individuals; it appears that this is a separate phenomenon to the change in diastolic filling. PMID- 15354034 TI - Thirst sensations and AVP responses at rest and during exercise-cold exposure. AB - PURPOSE: The purpose of this study was to 1) determine the effect of hypohydration (HYPO) on thirst sensations during moderate exercise in the cold and 2) determine a possible mechanism for a cold-induced decline in thirst. METHODS: In the first phase of the study, eight males walked on four occasions, in T-shirts, shorts, and shoes, at 50% VO2max, for 60 min in either a 4 degrees C (cold) or 27 degrees C (temperate) environment in a state of HYPO or euhydration (EU). In the second phase, nine males in states of EU or HYPO randomly performed four trials consisting of 30 min standing at 27 degrees C, followed by 30 min of standing and 30 min of treadmill exercise at 50% of VO2max, in either 4 degrees C or 27 degrees C air. RESULTS: In phase 1, thirst sensations were lower throughout 60 min of exercise (P < 0.05) in both HYPO and EU conditions during the cold trials. In phase 2, despite elevated plasma osmolality (P < 0.05), perception of thirst and plasma arginine vasopressin [AVP] similarly decreased (P < 0.05) after 30-min standing rest and 30-min exercise in the HYPO-cold trial, compared with the HYPO-temperate, EU-cold, and EU-temperate trials. CONCLUSIONS: When either euhydrated or hypohydrated, cold exposure attenuated thirst by up to 40% at rest and during moderate-intensity exercise. The attenuated thirst when hypohydrated during cold exposure was associated with lower plasma [AVP] despite an elevated plasma osmolality. This decline in thirst and AVP in the cold may be the result of peripheral vasoconstriction, mediating an increase in central blood volume and stimulation of central volume receptors. PMID- 15354035 TI - Saliva parameters as potential indices of hydration status during acute dehydration. AB - PURPOSE: Firstly, to identify whether saliva flow rate, osmolality, and total protein are potential markers of hydration, we compared changes in these parameters with changes in plasma osmolality during progressive dehydration. Secondly, we compared the sensitivity of saliva parameters to track hydration changes with the sensitivity of urine osmolality. Thirdly, to test the hypothesis that dehydration, rather than neuroendocrine regulation, is responsible for the decrease in saliva flow rate during prolonged exercise, we compared flow rate and catecholamine responses to prolonged exercise with and without fluids. METHODS: colon; Fifteen males (plasma osmolality 289 +/- 4 mOsmol x kg(-1); mean +/- SD) exercised (30 degrees C, 70% RH) with no fluid intake (NFI) until body mass loss (BML) of 1.1, 2.1, and 3.0% and on another occasion with fluid intake (FI) to offset losses. RESULTS: colon; Plasma and urine osmolality increased during NFI (plasma osmolality 3.0% BML: 298 +/- 4 mOsmol x kg(-1); P < 0.01). Saliva flow rate decreased (P < 0.01), saliva total protein increased (P < 0.01), and saliva osmolality increased from preexercise (50 +/- 11 mOsmol x kg(-1)) to 3.0% BML (105 +/- 41 mOsmol x kg(-1)) during NFI (P < 0.01). Saliva osmolality, urine osmolality, and saliva total protein correlated strongly with plasma osmolality during dehydration (r 0.87, 0.83, and 0.91, respectively; P < 0.01). During the FI trial, saliva flow rate and osmolality remained unchanged. Plasma catecholamine concentration increased during exercise (P < 0.01) with no difference between trials. CONCLUSIONS: colon; Saliva osmolality and total protein appear to be as sensitive as urine osmolality to track hydration changes during hypertonic-hypovolemia. These results also suggest that dehydration has a greater involvement in the decrease in saliva flow rate during prolonged exercise than neuroendocrine regulation. PMID- 15354036 TI - The effect of glucose infusion on glucose kinetics during a 1-h time trial. AB - PURPOSE AND METHODS: To investigate the effect of glucose infusion on glucose kinetics and performance, six endurance cyclists (VO2max = 61.7 +/- 2.0 (mean +/- SE) mL x kg(-1) x min(-1)) completed two performance trials in which they had to accomplish a set amount of work as quickly as possible (991 +/- 41 kJ). Subjects were infused with either glucose (20% in saline; carbohydrate (CHO)) at a rate of 1 g x min(-1) or saline (0.9% saline; placebo (PLA)). It was hypothesized that time trial performance would be unaffected by the infusion of glucose, as endogenous stores of CHO would not be limiting in the PLA trial. RESULTS: Plasma glucose concentration increased from 4.8 +/- 0.1 mmol x L(-1) to 5.9 +/- 0.3 mmol x L(-1) during the PLA trial and from 4.9 +/- 0.1 mmol x L(-1) at rest to 12.4 +/ 1.1 mmol x L(-1) during the CHO trial. These values were significantly higher at all time points during the CHO trial compared with PLA (P < 0.001). In the final stages of the time trial, Rd in the PLA trial was 49 +/- 5 micromol x kg(-1) x min(-1) compared with 88 +/- 7 micromol x kg(-1) x min(-1) in the CHO trial (P < 0.05). Despite these differences, there was no difference in performance time between PLA and CHO (60.04 +/- 1.47 min, PLA, vs 59.90 +/- 1.49 min, CHO, respectively). Infused carbohydrate oxidation in the last 25% of the CHO trial was at least 675 +/- 120 micromol x kg(-1) and contributed 17 +/- 4% to total carbohydrate oxidation. CONCLUSION: The results demonstrate that glucose infusion had no effect on 1-h cycle time-trial performance, despite an increased availability of plasma glucose for oxidation and evidence of increased glucose uptake into the tissues. PMID- 15354037 TI - High oxidation rates from combined carbohydrates ingested during exercise. AB - Studies that have investigated oxidation of a single carbohydrate (CHO) during exercise have reported oxidation rates of up to 1 g x min(-1). Recent studies from our laboratory have shown that a mixture of glucose and sucrose or glucose and fructose ingested at a high rate (1.8 g x min(-1)) leads to peak oxidation rates of approximately 1.3 g x min(-1) and results in approximately 20 to 55% higher exogenous CHO oxidation rates compared with the ingestion of an isocaloric amount of glucose. PURPOSE: The purpose of the present study was to examine whether a mixture of glucose, sucrose and fructose ingested at a high rate would result in even higher exogenous CHO oxidation rates (>1.3 g x min(-1)). METHODS: Eight trained male cyclists (VO2max: 64 +/- 1 mL x kg(-1) BM x min(-1)) cycled on three different occasions for 150 min at 62 +/- 1% VO2max and consumed either water (WAT) or a CHO solution providing 2.4 g x min(-1) of glucose (GLU) or 1.2 g x min(-1) of glucose + 0.6 g x min(-1) of fructose + 0.6 g x min(-1) of sucrose (MIX). RESULTS: High peak exogenous CHO oxidation rates were found in the MIX trial (1.70 +/- 0.07 g x min(-1)), which were approximately 44% higher (P < 0.01) compared with the GLU trial (1.18 +/- 0.04 g x min(-1)). Endogenous CHO oxidation was lower (P < 0.05) in MIX compared with GLU (0.76 +/- 0.12 and 1.05 +/- 0.06 g x min(-1), respectively). CONCLUSION: When glucose, fructose and sucrose are ingested simultaneously at high rates (2.4 g x min(-1)) during cycling exercise, exogenous CHO oxidation rates can reach peak values of approximately 1.7 g x min( 1) and estimated endogenous CHO oxidation is reduced compared with the ingestion of an isocaloric amount of glucose. PMID- 15354038 TI - Multivariate genetic analysis of lifetime exercise and environmental factors. AB - PURPOSE: We investigated whether the association between exercise and individual specific factors that correlate with exercise may be explained by genetic or common environmental factors. METHODS: Lifetime exercise data were available from 147 MZ and 153 DZ adult male twin pairs with a mean age of 50 yr (SD = 8 yr). RESULTS: The best-fitting quantitative genetic model for adulthood exercise level consisted of additive genetic effects, genetic effects due to dominance and unique environment effects, with genetic effects explaining 51% (95% CI = 29-63%) of the variance. Factors associated with adulthood exercise level were adolescent exercise, participation in competitive sports, perceived health, smoking status, and percent body fat. In bivariate models, approximately half of the covariation between those factors and adulthood exercise level was accounted for by unique environmental effects (i.e., factors not shared by the co-twins). Additive genetic effects explained less (3-20%) than dominance genetic effects (23-53%) of the covariation between those factors and adulthood exercise. Shared environmental effects were present only in the bivariate model of adulthood and adolescent exercise, explaining 11% of the covariance. CONCLUSIONS: : The genetic component shared in common by exercise and factors associated with exercise suggests that there may be a complex pathway of genetic selection and predisposition for a physically active lifestyle. PMID- 15354039 TI - Descriptive epidemiology of pedometer-determined physical activity. AB - PURPOSE: The dual purposes of this study were: 1) to provide preliminary descriptive epidemiology data representing pedometer-determined physical activity (PA) and 2) to explore sources of intra-individual variability in steps per day. METHODS: All participants (76 males, age = 48.4 +/- 16.3 yr, body mass index (BMI) = 27.1 +/- 5.1 kg x m(-2); 133 females, age = 47.4 +/- 17.5 yr, BMI = 26.9 +/- 5.7 kg x m(-2)) resided in Sumter County, SC, and were recruited by telephone to receive a mailed kit to self-monitor PA for 1 wk. Statistical analyses compared mean steps per day between sexes, races, age groups, education and income levels, and BMI categories. Mean steps per day were also compared between: 1) weekdays versus weekend days, 2) workdays versus nonworkdays, and 3) days of sport/exercise versus no participation. RESULTS: The entire sample took 5931 +/- 3664 steps x d(-1) (males = 7192 +/- 3596 vs females = 5210 +/- 3518 steps x d( 1), t = 7.88, P < 0.0001). Significant differences were also indicated by race, age, education, income, and BMI. In addition, weekdays were significantly higher than weekend days, workdays were higher than nonworkdays, and sport/exercise days were higher than nonsport/exercise days. CONCLUSIONS: The large standard deviations reflect a wide distribution of ambulatory behavior. Regardless, important differences are still evident by demographic characteristics, BMI categories, day of the week, and reported engagement in work or sport/exercise. PMID- 15354040 TI - Reliability of motor-evoked potentials during resting and active contraction conditions. AB - PURPOSE: To determine the reliability of motor-evoked potentials (MEP) obtained using transcranial magnetic stimulation (TMS) in the first dorsal interosseous (FDI) and biceps brachii muscles. METHODS: Fourteen college subjects attended the laboratory on three separate days. TMS was used to obtain MEP with the subject relaxed (resting condition) at stimulation intensities of 70%, 85%, and 100% of maximal stimulator output. MEP were also obtained during four active contraction conditions involving contractions of 25%, 50%, 75%, and 100% of maximal effort (MVC). Reliability was measured using an intraclass correlation analysis of variance (ANOVA) design. RESULTS: In the resting condition, substantial increases in MEP amplitude were observed for both muscles from day 1 to day 2. Intraclass reliability estimates were higher for the biceps muscle (ICC = 0.95-0.99) than for the FDI muscle (ICC = 0.60-0.81). During the active conditions, the greatest MEP were observed at 25% and 50% MVC, with smaller MEP at 75% and 100% MVC. Intraclass correlations in the active condition were approximately 0.63-0.73. CONCLUSIONS: : Moderate to good reliability of MEP amplitude in the biceps and FDI muscles can be obtained using TMS in both resting and active contraction conditions. PMID- 15354041 TI - Muscle plasticity after weight and combined (weight + jump) training. AB - PURPOSE: The purpose of this study was to compare the effects of jump training as a complement to weight training on jump performance and muscle strategy during the squat and countermovement jump. METHOD: Twenty-two male handball players, between the ages of 17 and 24, and in good health, were randomly divided into three groups. Two were trained groups, weight training (WTG) and jump training combined with weight training (CTG), and one was a control group (CG). Maximal isometric force and maximal concentric power were assessed by a supine leg press, squat jump (SJ), counter movement jump (CMJ), and surface EMG was used to determine changes in muscle adaptation before and after the training period. RESULTS: After 6-wk training programs, the two training groups increased maximal isometric force, maximal concentric power, and squat jump performance. However, only combined training presented a significant increase in height jump performance during the countermovement jump (P < 0.05). EMG analysis (as interpreted through the root mean square values) showed that the SJ was performed similarly before and after the training period for the two training groups. However, during the CMJ, only the CTG group adopted a new technique manifested by a short transition phase together with an increase in knee joint stiffness and knee extensor muscle activation and rectus femoris ratio. CONCLUSION: It was suggested that the central activities in knee joint during the transition phase, in conjunction with intrinsic muscle contractile properties, play a major role in the regulation of performance during a CMJ. Furthermore, our study suggests that a change in maximal strength and/or explosive strength does not necessarily cause changes in combined movement such as the stretch shortening cycle. PMID- 15354042 TI - Evaluation of power prediction equations: peak vertical jumping power in women. AB - PURPOSE: The purpose of this investigation was to: 1) compare actual peak power (PPactual) to estimated values (PPest) derived from three different prediction equations (Sayers and Harman), 2) determine the ability of the prediction formulas to monitor change following 6 wk of plyometric training, and 3) generate a new regression model. METHODS: colon; Twenty college females (age = 20.1 +/- 1.6 yr; body mass = 65.9 +/- 8.9 kg) were randomly assigned to a control or intervention group. Pre- and posttest countermovement jump (CMJ) height and PPactual were determined simultaneously on a force platform. Body mass and maximal CMJ height were used to predict peak power. RESULTS: colon; All three PPest were significantly correlated 0.84-0.99) and post (r = 372.4 W) was significantly less to PPactual and to each other on pre (r = 0.88-0.99) tests. PPactual (2425.4 +/- 2920.8 +/- 482.6 W; CMJ = 2925.1 +/- 409.7 than PPest (Sayers: SJ = 473.0 W) but was not different from PPest (Harman: 2585.0 +/- 409.7 W). Posttests revealed similar differences between PPactual and PPest for the intervention group, however no significant differences were observed for the control group. Mean differences from pre and posttests did not differ within or between PPest. Regression analysis determined the formula: ppest = 65.1 x (jump height) + 25.8 x (body mass) - 1413.1 (R = 0.92; SEE = 120.8), which slightly determined (0.77%) peak power is compared with PPactual in our cross-validation sample (n = 7) CONCLUSIONS: colon; Changes in peak power is accurate using any of the regression equations; however, the new prediction formula and that of Harman seem to more precisely estimate peak power. Strict jumping technique along with simultaneous measurement of power and jump height should be used as the standard for comparison. PMID- 15354043 TI - Bone mineral density of olympic-level female winter sport athletes. AB - PURPOSE: To compare areal bone mineral density (aBMD) of female winter sport athletes to healthy controls of similar age and body mass index (BMI). METHODS: Areal BMD (g x cm(-2)) of the whole body, lumbar spine (L2-L4), and right proximal femur were assessed by dual energy x-ray absorptiometry in athletes (N = 40; age: 26.1 +/- 5.7 yr; ht: 165.6 +/- 0.1 cm; wt: 63.0 +/- 6.5 kg; BMI: 23.0 +/ 1.9 kg x m(-2)) involved in speed skating (N = 9), snowboarding (N = 13), freestyle skiing (N = 3), biathlon (N = 8), bobsleigh, skeleton, luge (N = 7), and controls (N = 21; age: 26.0 +/- 5.1 yr; ht: 165.8 +/- 0.1 cm; wt: 62.8 +/- 5.9 kg; BMI: 22.9 +/- 1.3 kg x m(-2)). RESULTS: Using independent t-tests, athletes had lower fat mass, percent body fat, and higher lean mass than controls (P < 0.001). Areal BMD was higher in athletes than controls for all skeletal sites (P /= six observations for peak aerobic power to estimate age at peak velocity (PV) for peak VO2 (age at PVPVO2), PVPVO2 (L x min(-1) x yr(-1)), and value at PVPVO2 (L x min(-1)) for each individual. Curves were successfully fitted for 83 individuals (48 males, 35 females). The model was also fitted to individual data for height and weight to estimate ages at peak height velocity (PHV) and peak weight velocity (PWV). Age at PVPVO2 was compared with ages at PHV and PWV. Pearson correlation coefficients were calculated between ages at PV and PV for peak VO2, height, and weight. RESULTS: Mean ages at PVPVO2 are 12.3 +/- 1.2 yr for females and 14.1 +/- 1.2 yr for males. Peak VO2 increases in both sexes throughout adolescence, with males having higher values than females at all ages. Age at PVPVO2 occurs nearly coincident with PHV and before PWV in both sexes. Correlation coefficients among ages at PHV, PWV, and PVPVO2 suggest a general maturity factor for body size and aerobic power. CONCLUSION: Growth in peak VO2 exhibits a clear growth spurt in both sexes during adolescence. The growth spurt occurs earlier in females but is of greater magnitude in males. PMID- 15354047 TI - Prediction of activity energy expenditure using accelerometers in children. AB - PURPOSE: To validate two accelerometer-based activity monitors as measures of children's physical activity using energy expenditure as the criterion measure. METHODS: Actiwatch (AW) and Actical (AC) activity monitors were validated against continuous 4-h measurements of energy expenditure (EE) in a respiratory room calorimeter and 1-h measurements in an exercise laboratory using a portable calorimeter and treadmill in 32 children, ages 7-18 yr. The children performed structured activities including basal metabolic rate (BMR), playing Nintendo, using a computer, cleaning, aerobic exercise, ball toss, treadmill walking, and running. Equations were developed to predict activity energy expenditure (AEE = EE - BMR), and physical activity ratio (PAR = EE/BMR) from a power function of AW or AC, and age, sex, weight, and height. Thresholds were determined to categorize sedentary, light, moderate, and vigorous levels of physical activity. RESULTS: Activity counts accounted for the majority of the variability in AEE and PAR, with small contributions of age, sex, weight, and height. Overall, AW equations accounted for 76-79% and AC equations accounted for 81% of the variability in AEE and PAR. Relatively wide 95% prediction intervals suggest the accelerometers are best applied to groups rather than individuals. Sensitivities were higher for the vigorous threshold (97%) than the other thresholds (86-92%). Specificities were on the order of 66-73%. The positive predictive values for sedentary, light, moderate, and vigorous categories were 80, 66, 69, and 74% for AW, respectively, and 81, 68, 72, 74% for AC, respectively. CONCLUSION: Both accelerometer-based activity monitors provided valid measures of children's AEE and PAR, and can be used to discriminate sedentary, light, moderate, and vigorous levels of physical activity but require further development to accurately predict AEE and PAR of individuals. PMID- 15354048 TI - Consistency of the talk test for exercise prescription. AB - INTRODUCTION/PURPOSE: The Talk Test has been shown to be well correlated with the ventilatory threshold, with accepted guidelines for exercise prescription, and with the ischemic threshold. As such, it appears to be a valuable although quite simple method of exercise prescription. In this study, we evaluate the consistency of the Talk Test by comparing responses during different modes of exercise. METHODS: Healthy volunteers (N = 16) performed incremental exercise, on both treadmill and cycle ergometer. Trials were performed with respiratory gas exchange and while performing the Talk Test. Comparisons were made regarding the correspondence of the last positive, equivocal, and first negative stages of the Talk Test with ventilatory threshold. RESULTS: The %VO2peak, %VO2 reserve, %HRpeak, and %HR reserve at ventilatory threshold on treadmill versus cycle ergometer (77%, 75%. 89%, and 84% vs 67%, 64%, 82%, and 74%) were not significantly different than the equivocal stage of the Talk Test (83%, 82%, 86%, and 80% vs 73%, 70%, 87%, and 81%). The VO2 at ventilatory threshold and the last positive, equivocal and negative stages of the Talk Test were well correlated during treadmill and cycle ergometer exercise. CONCLUSIONS: The results support the hypothesis that the Talk Test approximates ventilatory threshold on both treadmill and cycle. At the point where speech first became difficult, exercise intensity was almost exactly equivalent to ventilatory threshold. When speech was not comfortable, exercise intensity was consistently above ventilatory threshold. These results suggest that the Talk Test may be a highly consistent method of exercise prescription. PMID- 15354049 TI - Reliability of accelerometry-based activity monitors: a generalizability study. AB - INTRODUCTION: Numerous studies have examined the validity of accelerometry-based activity monitors but few studies have systematically studied the reliability of different accelerometer units for assessing a standardized bout of physical activity. Improving understanding of error in these devices is an important research objective because they are increasingly being used in large surveillance studies and intervention trials that require the use of multiple units over time. METHODS: Four samples of college-aged participants were recruited to collect reliability data on four different accelerometer types (CSA/MTI, Biotrainer Pro, Tritrac-R3D, and Actical). The participants completed three trials of treadmill walking (3 mph) while wearing multiple units of a specific monitor type. For each trial, the participant completed a series of 5-min bouts of walking (one for each monitoring unit) with 1-min of standing rest between each bout. Generalizability (G) theory was used to quantify variance components associated with individual monitor units, trials, and subjects as well as interactions between these terms. RESULTS: The overall G coefficients range from 0.43 to 0.64 for the four monitor types. Corresponding intraclass correlation coefficients (ICC) ranged from 0.62 to 0.80. The CSA/MTI was found to have the least variability across monitor units and trials and the highest overall reliability. The Actical was found to have the poorest reliability. CONCLUSION: The CSA/MTI appeared to have acceptable reliability for most research applications (G values above 0.60 and ICC values above 0.80), but values with the other devices indicate some possible concerns with reliability. Additional work is needed to better understand factors contributing to variability in accelerometry data and to determine appropriate calibration protocols to improve reliability of these measures for different research applications. PMID- 15354050 TI - Evaluation of the 7-day physical activity recall in urban and rural men. AB - PURPOSE: To assess reliability and validity of the 7-d physical activity recall (PAR) in 60- to 80-yr-old men from urban and rural homes in the southeastern United States. METHODS: Two hundred twenty 60- to 80-yr-old men (29% minority ethnicity) from Veterans Affairs primary care clinics completed two PAR interviews at 2- to 4-wk intervals. The PAR included modifications to elicit and score light-moderate (2-3 METs) as well as moderate (3-6 METs) and hard (>/=7 METs) physical activity. Reliability was assessed using intraclass correlations from the two interviews. Validity was assessed using Spearman correlations of PAR variables with 6-min walk performance, and with treadmill performance and accelerometer activity counts from subsamples, as well as measures of gait, balance, activities of daily living, and perceived quality of life. Results were examined separately for urban and rural participants. RESULTS: Reliability coefficients were acceptable and PAR measures of estimated energy expenditure, and time spent in hard, moderate, and light-moderate activity correlated as hypothesized with the fitness and health measures. CONCLUSION: Reliability and validity coefficients were generally similar to those found in studies of the PAR with younger samples and with instruments designed specifically for use with elders. The PAR is a useful and flexible instrument for assessing physical activity of varying intensity in older men living in both urban and rural home settings. PMID- 15354051 TI - Pneumatic resistance machines can provide eccentric loading. PMID- 15354052 TI - Pneumatic resistance machines can provide eccentric loading: response. PMID- 15354053 TI - Respirable particles and carcinogens in the air of delaware hospitality venues before and after a smoking ban. AB - How do the concentrations of indoor air pollutants known to increase risk of respiratory disease, cancer, heart disease, and stroke change after a smoke-free workplace law? Real-time measurements were made of respirable particle (RSP) air pollution and particulate polycyclic aromatic hydrocarbons (PPAH), in a casino, six bars, and a pool hall before and after a smoking ban. Secondhand smoke contributed 90% to 95% of the RSP air pollution during smoking, and 85% to 95% of the carcinogenic PPAH, greatly exceeding levels of these contaminants encountered on major truck highways and polluted city streets. This air-quality survey demonstrates conclusively that the health of hospitality workers and patrons is endangered by tobacco smoke pollution. Smoke-free workplace laws eliminate that hazard and provide health protection impossible to achieve through ventilation or air cleaning. PMID- 15354054 TI - Respiratory symptoms and lung function of coke oven workers: a lung function surveillance system from 1990-2000. AB - This study investigated the association of exposure to coke oven emissions with respiratory symptoms and with lung function. Data for respiratory symptoms and lung function were repeatedly collected from 764 male coke oven workers between 1990 and 2000. The data were analyzed cross-sectionally and longitudinally. Working in "Operation" increased the risk for cough (odds ratio [OR] = 2.37), phlegm (OR = 2.55), and shortness of breath (OR = 1.52). Cross-sectionally, each year of working in Operation was associated with a reduction in FEV1 of approximately 10 mL (P = 0.006) and in FVC of around 21 mL (P = 0.0001). From longitudinal analysis, each year of working in Operation was associated with an extra annual decline in FEV1 of 1.5 mL (P = 0.0012) and in FVC of around 1.8 mL (P = 0.0017). Exposure to coke oven emissions increased the risk for respiratory symptoms and adversely affected lung function. PMID- 15354055 TI - A population-based study on the association between parental occupations and some common birth defects in singapore (1994-1998). AB - We sought to study the association between some common birth defects and parental occupations. The live births and congenital malformation born between January 1994 and December 1998 were obtained from the Singapore National Registry of Births and Deaths (the National Birth Defect Registry). The prevalence of overall birth defects among Singapore live births during the study period is 13.9 per 1,000 live births. The most frequent single coding defects were "bulbus cordis anomalies and anomalies of cardiac septal closure (BCA)," "congenital anomalies of urinary system (CUS)," "cleft palate and cleft lip (CPL)," and "certain congenital musculoskeletal deformities (CMD)." Using the "Legislators, Senior Officers & Managers" as reference and adjusting for possible confounders, there were significant associations for: (1) paternal "clerical workers" (adjusted RR 2.25) with the BCA; (2) maternal "professionals" with CUS (adjusted risk ratio [RR] 3.58); (3) paternal "production craftsmen and related workers" with both the BCA (adjusted RR 2.04), and the CMD (adjusted RR 2.83); (4) paternal "plant and machine operators and assemblers" with the BCA (adjusted RR 2.49), and the CUS (adjusted RR 5.19), and the CMD (adjusted RR 3.01). Paternal rather than maternal exposure might be more important in the causation of some common birth defects in Singapore. PMID- 15354056 TI - A prospective study of prenatal and childhood lead exposure and erythropoietin production. AB - We test the hypothesis that chronic lead (Pb) exposure may be associated with an inability to maintain an adequate serum erythropoietin (EPO) concentration. From a longitudinal study of Pb exposure and infant and childhood development, we measured blood Pb (BPb) and serum EPO concentrations serially at ages 4.5, 6.5, 9.5, and 12 and tibia (cortical) Pb concentration at age 12. Pb-exposed children aged 4.5 and 6.5 produced increased concentrations of EPO to maintain normal Hgb concentrations. EPO production declined between ages 4.5 and 6.5. At ages 9.5 and 12, further diminution of the association was found. No association was found between tibia Pb and EPO. The continued decline in the slope of the relationship between EPO and BPb with age, after adjustment for hemoglobin, implies a gradually decreasing capacity to produce EPO. PMID- 15354058 TI - Biological monitoring of exposures to aluminium, gallium, indium, arsenic, and antimony in optoelectronic industry workers. AB - The main objective of this study was to investigate aluminum, gallium, indium, arsenic, and antimony exposures on blood and urine levels in the optoelectronic workers. One hundred seventy subjects were enrolled in this cohort study. Whole blood and urine levels were determined by inductively coupled plasma-mass spectrometry. Blood indium and urine gallium and arsenic levels in the 103 workers were significantly higher than that in 67 controls during the follow-up period. In regression models, the significant risk factors of exposure were job title, preventive equipment, Quetelet's index, sex, and education level. The findings of this study suggest that gallium, indium, and arsenic exposure levels may affect their respective levels in blood and urine. The use of clean, preventive equipment is recommended when prioritizing the administration of safety and hygiene in optoelectronics industries. PMID- 15354059 TI - Comparing excess costs across multiple corporate populations. AB - The purpose of this study was to examine the relationship of health risk level to charged medical costs and determine the excess cost of higher risk individuals compared to low risk. Two years of medical claims from six corporations were used to determine costs of health risk assessment (HRA) participants and nonparticipants. A total of 165,770 employees, 21,124 of which took an HRA, were used for the study. Costs increased as risk level increased. There were no significant differences within a risk level between companies for the cost ratio. Percent of medical costs due to excess risk ranged from 15.0-30.8% for HRA participants and 23.8-38.3% for the study population. Cost patterns were consistent across companies. Excess cost as the result of increased risk level accounted for a substantial portion of the cost at each company. These results can be used to justify the need for a health-promotion program and to estimate potential savings as the result of excess risk. Even without the use of an HRA, health practitioners should feel confident stating that excess risk accounts for at least 25% to 30% of medical costs per year across a wide variety of companies, regardless of industry or demographics. The numbers can be used as a realistic estimate for any health promotion program financial proposal. PMID- 15354060 TI - Impact of air pollution on cardiopulmonary fitness in schoolchildren. AB - We studied the association between air pollution and cardiopulmonary fitness among 821 schoolchildren aged 8 to 12 from two districts with different air quality in Hong Kong. Their parents completed a respiratory questionnaire, and the maximum oxygen uptake (VO2max) of the children was assessed using the multistage fitness test. After adjusting for potential confounding factors, children in the high pollution district had a significantly lower VO2max than those in the low pollution district (27.9 mL.kg(-1).min(-1) vs. 29.8 mL.kg( 1).min(-1)). Habitual physical exercise was associated with a higher VO2max in the low-pollution district but not in the high-pollution district. Air pollution adversely affected the VO2max in children, and physical exercise in a polluted environment might not have beneficial effect on cardiopulmonary fitness. PMID- 15354061 TI - Beryllium medical surveillance at a former nuclear weapons facility during cleanup operations. AB - Despite increasing need to remediate beryllium-contaminated buildings in industry, little is known about the magnitude of risk associated with beryllium abatement or the merits of beryllium medical surveillance for cleanup workers. We examined beryllium lymphocyte proliferation tests and reviewed medical evaluations on workers at a nuclear weapons facility during the process of decontamination and decommissioning. Of 2,221 workers, 19 (0.8%) were beryllium sensitized based on two or more abnormal beryllium lymphocyte proliferation tests. Eight of 19 sensitized individuals underwent full clinical evaluation, of whom two were diagnosed with chronic beryllium disease (CBD). Notably, seven beryllium sensitized and CBD cases were hired after the start of cleanup operations. Beryllium medical surveillance detects sensitization and CBD in cleanup workers. Exposure controls and medical surveillance need to be 'broad based' to include all cleanup workers involved in beryllium-contaminated building remediation. PMID- 15354062 TI - Liver fibrosis in asymptomatic polyvinyl chloride workers. AB - This study was designed to determine whether vinyl chloride monomer (VCM) exposure is associated with liver fibrosis. A total of 347 workers with occupational exposure to VCM were systemically examined using liver ultrasonography and routine liver function tests. Vinyl chloride monomer cumulative dose (ppm-month) was estimated by summing the products of air VCM concentration levels and months of employment. Liver fibrosis was defined in subjects with precirrhosis and cirrhosis of liver diagnosed using ultrasonography. Significantly increased risks of developing liver fibrosis were found in workers who had history of high exposure jobs (odds ratio 5.5, 95% confidence interval 1.7-25.4) when compared with workers who did not have history of high exposure jobs. We concluded that there was an increased risk of developing liver fibrosis in PVC workers who had high exposure to VCM. PMID- 15354063 TI - Current pediatric and maternal lead levels in blood and breast milk in Andean inhabitants of a lead-glazing enclave. AB - Prenatal and postnatal lead (Pb) exposure may induce neurodevelopmental disabilities in children. As part of an ongoing health-monitoring study, blood lead (PbB) levels were compared in 90 children tested in 2003 (current group) and 166 children tested between 1996 and 2000 (reference group) in Ecuadorian Andean villages with high Pb contamination. The mean PbB level for children in the reference group was 40 microg/dL (range, 6.2-119.1), and significantly higher than the mean PbB level of 25.5 microg/dL (range, 2.1-94.3) for the current group (t test, P = 0.0001). An analysis of variance revealed no significant main effects for age and gender and no significant interaction between age and gender for the current group but a significant age by gender interaction for the reference group (F = 5.96, P = 0.01). Regression analysis revealed a significant correlation (r = 0.258, P = 0.01) between PbB level and age for males but not for females in the reference group. The Pb levels in breast milk from nursing mothers ranged from 0.4-20.5 microg/L (mean, 4.6), and the PbB levels in the breastfeeding mothers ranged from 4.5-35.2 microg/dL (mean, 17.1). The PbB levels of mother-infant pairs ranged from 4.6-27.4 microg/dL for mothers and 3.9-33.5 microg/dL for infants. The results showed significantly reduced PbB levels in children in the study area and suggest that a Pb education and prevention program contributed to the current reduction in Pb intoxication. PMID- 15354064 TI - Occupational injury surveillance among electric utility employees. AB - Currently, there are only limited injury surveillance data for the electric utility workforce. To address this gap, an Occupational Health Surveillance Database for electric power utilities was established for epidemiologic monitoring and intervention program evaluation. Injury rates varied across utility occupations, such as, managers, line workers, and meter readers, ranging from 0.18 to 9.63 per 100 employee-years based on more than 500,000 employee years of observation. Compared with male workers, the risk of injury among female workers was lower overall, although their risk was higher in nonoffice occupations than their male counterparts. Across the period 2000 to 2002, three of four companies that experienced decreases in workforce size also experienced noticeable increases in injury rates. Our results suggest that benchmarking and prevention efforts should be directed at specific occupational groups and specific injury types. PMID- 15354065 TI - A case-control study of parental occupation, leukemia, and brain tumors in an industrial city in Taiwan. AB - We conducted a case-control study in an industrial city in Taiwan to determine whether parents of newly diagnosed patients who were younger than 30 years old with leukemia or brain tumors or the patients themselves were more likely to have been employed in certain occupations or industries. Job histories were collected for parents (and for subjects if they worked) on 103 newly diagnosed cases of leukemia, 74 newly diagnosed cases of brain tumors, and 417 controls matched for age and sex. All jobs since the age of 16 that the subjects held for more than 6 months, a total of approximately 4,000 jobs, were coded for occupation and industry according the standard four-digit system used in Taiwan. Matched-pair analyses were performed comparing cases and controls among all jobs held by subjects and both parents using four-digit occupation and industry codes. Separate analyses also were performed for parental jobs held during the preconception, perinatal, and postnatal periods. Odds ratios (ORs) were adjusted for subject smoking, parental smoking, and exposure to medical radiation. Certain industry and occupation four-digit codes were significantly associated with increased odds ratios of childhood tumors. Including work during any or all periods, leukemias were more common in children of fathers who had worked (1) as wood treaters (adjusted OR 16.03, 95% confidence interval CI = 1.77-145.5), and (2) as building finishers and related trades workers (adjusted OR 4.08, 95% CI = 1.12-14.8), whereas brain tumors were more common in children of mothers who had worked (1) in electronic parts and components manufacturing (adjusted OR 13.78, 95% CI = 1.47-129.0) and 2) as textile and garment workers (adjusted OR 7.25, 95% CI = 1.42-37.0), as well as in subjects who had worked with certain electronic parts and components (adjusted OR 28.67, 95% CI = 2.88-285.6). Leukemias were more common in children of fathers who had worked in the preconception period (1) as wood treaters (adjusted OR 12.17, 95% CI = 1.36-109.2), (2) as building finishers and related trades workers (adjusted OR 4.08, 95% CI = 1.12-14.8), (3) as electronic equipment assemblers (adjusted OR 4.56, 95% CI = I 1.05-19.9), and (4) as certain other assemblers (adjusted OR 10.24, 95% CI = 1.02-102.6). In addition, leukemias were more common in children of fathers who had worked in the perinatal period (1) as wood treaters (adjusted OR 13.08, 95% CI = 1.36-125.5) and (2) as building finishers and related trades workers (adjusted OR 4.51, 95% CI = 1.04-19.6). Brain tumors were more common in children of mothers who had worked in the preconception period (1) in electronic parts and components manufacturing (adjusted OR 11.81, 95% CI = 1.20-116.3), and (2) as textile and garment workers (adjusted OR 7.25, 95% CI = 1.18-31.0). PMID- 15354066 TI - Psychosocial factors and low back pain, consultations, and sick leave among farmers and rural referents: a population-based study. AB - Farmers have more low back pain (LBP) than nonfarmers. In a previous report, we found that differences between farmers and nonfarmers in physical work exposure did not explain the LBP differences. In this report, we tested the hypothesis that psychosocial factors might explain the differences in LBP reporting, medical consultation, and sick leave. A cross-sectional population-based survey of 1,013 middle-aged farmers and 769 matched referents was performed. Data on LBP, consultations, and sick leave during lifetime was obtained along with information on psychosocial, social network, and lifestyle variables. Several of the psychosocial variables were associated with LBP but the difference in LBP prevalence between farmers and nonfarmers could be explained only marginally. Farmers and self-employed referents tended to have lower odds of sick leave because of LBP than employed referents after adjustment for psychosocial factors. PMID- 15354070 TI - Reversal of disc cupping after intraocular pressure reduction in topographic image series. AB - PURPOSE: To identify and characterize 'reversal' of optic nerve cupping following intraocular pressure (IOP) lowering in scanning laser tomography (SLT) longitudinal image series. METHODS: Modification was made to a previously described analytical approach to longitudinally study putatively increased rim area following IOP lowering. Sustained IOP reduction of 25% was by topical medication. Forty SLT image series with equivalent follow up were assessed: 10 with ocular hypertension (OHT), 10 with primary open angle glaucoma (POAG), and as controls, 20 normal. Reproducible rim area reversal was identified by sector and its time-course over 1 year examined. RESULTS: By a 2-of-3 reproducibility criterion, reversal following IOP lowering was confirmed in about a third of treated eyes (POAG and OHT) but not in any controls. Rim sectors showing reversal were mostly nasal, with a few occurring superotemporally. Reversal in a fifth of treated eyes persisted for at least 1 year; all these were in the nasal half of the disc. The number of sectors with persisting reversal affected less than 6% of all treated eyes' rim sectors. CONCLUSION: Rim area is not uncommonly increased after IOP lowering and this 'reversal' may persist for at least a year. Within topically treated eyes having IOP lowering of at least 25%, the proportion of rim sectors with persistent reversal appears small. Nevertheless, the effects of IOP reduction on topography, especially in the short term, should be considered when longitudinally assessing progressive rim loss in SLT images. PMID- 15354071 TI - Optic disc hemorrhages detected in a large-scale eye disease screening project. AB - PURPOSE: To investigate the prevalence and characteristics of optic disc hemorrhages. MATERIALS AND METHODS: We took IMAGEnet fundus photographs of 14,779 participants, aged 40 years or older in a large-scale eye disease screening project conducted in Tajimi, Japan. A single researcher reviewed all of the photographs for the presence of ocular abnormality in the optic nerve head and retina, paying special attention to the presence or absence of optic disc hemorrhages. Glaucomatous optic neuropathy was diagnosed based on the presence of nerve fiber layer defects and/or the appearance of the optic disc. RESULTS: Fundus photographs of 13,965 cases (27,930 eyes) were successfully reviewed bilaterally. We found disc hemorrhages at 92 locations of 92 (0.3%) eyes of 88 (0.6%) cases. Twenty-four hemorrhages of 23 (26%) cases were found in bilateral non-glaucoma cases and the remaining 68 hemorrhages of 65 (74%) cases were found in glaucoma cases. The prevalence of disc hemorrhages was 8.2% in 793 glaucoma cases and 0.2% in non-glaucoma cases. Hemorrhages developed significantly more frequently in females and in the elderly. The hemorrhages were predominantly found in the inferotemporal and superotemporal regions. The intraocular pressure was less than 20 mm Hg in all cases. The mean intraocular pressure was 15.0 mm Hg for glaucoma cases with hemorrhages and 13.9 mm Hg for non-glaucoma cases with hemorrhages. CONCLUSION: Disc hemorrhages occur more frequently in females, in elderly persons, and in glaucoma cases in Japanese aged 40 or older. The intraocular pressure of the eyes with disc hemorrhages is close to that of the normal population. PMID- 15354072 TI - Association between ocular herpes simplex virus and topical ocular hypotensive therapy. AB - PURPOSE: To evaluate whether use of particular topical hypotensive therapies is associated with ocular herpes simplex virus (OHSV). MATERIALS AND METHODS: This population-based, retrospective, cohort study used claims records from the Protocare Sciences managed care database (United States). Data were extracted from September 1, 1996 through June 30, 2002. An OHSV event was either a medical claim from the utilization database coded with an International Classification of Diseases Ninth Revision (ICD-9) code for OHSV (ICD-9 = 054.4, 054.40-054.44, 054.49) or a pharmacy claim for vidarabine or trifluridine ophthalmic solution. RESULTS: A total of 93,869 eligible glaucoma patients, 21 different ocular hypotensive agents, and 192,840 agent-utilizing patient combinations were identified. In all, 411 patients had an OHSV event; 272 of 411 patients had at least 1 ocular hypotensive agent dispensed prior to the OHSV event but not preceding the event by less than 7 days. Of these, 219 had only 1 ocular hypotensive agent dispensed on the last fill date prior to the OHSV event, yielding an overall OHSV event rate of 0.11%. There was no significant association between OHSV event rates and agent use for either the set of 21 agents (P = 0.260; chi2) or when 14 products having < 5% usage were combined (P = 0.058). Prevalence rates were estimated as 161 per 100,000 population for 2000 to 2001 and 165 per 100,000 for 1999 to 2001. CONCLUSIONS: Ocular herpes simplex virus is extremely rare in patients treated with ocular hypotensive therapies, and its prevalence is similar to that found in the general population. The current analysis revealed no association between the use of particular topical ocular hypotensive therapies and OHSV. PMID- 15354073 TI - Aspirin use in advanced uncontrolled glaucoma. AB - PURPOSE: To evaluate if aspirin use affects progression of primary open angle glaucoma (POAG). METHODS: A retrospective review of patients with uncontrolled glaucoma was performed. Incidence of aspirin use was noted by a one-time self reporting survey. Controls were medically stable patients diagnosed with POAG. The primary outcome measure studied was a comparison of percentages of aspirin use in patients who have and have not undergone glaucoma filtering surgery (trabeculectomy). RESULTS: Forty-one percent (26/64) of the patients in the trabeculectomy group and 23% (17/74) of controls were using aspirin. Patients undergoing trabeculectomy were twice as likely to take aspirin (O.R., 2.29; 95% C.I., 1.10-4.79). Subgroup analyses demonstrated increased aspirin use in those operative patients who are current or former smokers (O.R., 3.71; 95% C.I., 1.10 12.56), have systemic hypertension (O.R., 3.30; 95% C.I., 1.02-22.58), or have joint disease (O.R., 4.60; 95% C.I., 1.34-15.82). CONCLUSION: A higher concurrence of aspirin use was observed in patients with POAG who required surgical management compared with patients having relatively medically stable glaucoma. This may be secondary to a higher rate of glaucoma surgery performed on patients with greater systemic illnesses, more of whom use aspirin. PMID- 15354074 TI - Prevalence of primary open-angle glaucoma in a Spanish population: the Segovia study. AB - PURPOSE: To determine the prevalence of primary open-angle glaucoma (POAG) in Segovia, Spain. METHODS: We conducted a cross-sectional, population-based epidemiologic study, the target population of which was residents of Segovia, Spain, aged 40 to 79 years. A sample of 569 subjects was randomly selected in a stratified manner according to gender and age groups. All participants underwent a complete ophthalmic examination that included measurement of visual acuity and refraction, tonometry, anterior segment biomicroscopy, funduscopy, stereoscopic photographs of the optic nerve head, and automated white-on-white visual field testing. Two independent observers evaluated the optic nerve photographs and visual fields. The diagnosis of POAG was established when any eye had an open angle and a glaucomatous optic nerve and glaucomatous visual field. The prevalence of POAG in the population was estimated from the prevalence in the complete sample and the patients already diagnosed at the only glaucoma service in the city. RESULTS: The estimated prevalences (99% confidence interval) in the population were, respectively, 2.1% (1.9-2.3%), and 1.7% (1.6-1.8%) for POAG and ocular hypertension. The prevalence of POAG increased with age (P < 0.005) and tended to be greater (P = 0.054) in men (2.4%) than women (1.7%). CONCLUSION: The prevalence of POAG in this Segovia population is 2.1%, similar to that estimated in previous studies performed in predominantly Caucasian populations. PMID- 15354075 TI - Polymorphisms in the myocilin promoter unrelated to the risk and severity of primary open-angle glaucoma. AB - PURPOSE: To investigate the proximal 2.5 kb promoter in the myocilin (MYOC) gene for mutations in Chinese patients with primary open-angle glaucoma (POAG). PATIENTS AND METHODS: We screened for sequence alterations in the MYOC promoter in 88 unrelated Chinese patients with POAG and 94 unrelated individuals without glaucoma, aged 50 years or above, as control subjects. In addition, the specific MYOC.mt1 polymorphism was determined in a total of 212 POAG patients and 221 control subjects. The relationships between POAG phenotype and the identified polymorphisms were studied by univariate analysis, multivariable logistic regression analysis, and haplotype analysis. RESULTS: All polymorphisms identified in this study followed Hardy-Weinberg equilibrium (P > 0.12) both in POAG patients and controls. Both univariate and multivariable logistic regression analyses showed no polymorphism that was significantly associated with the risk of POAG, P > 0.08 and P > 0.044 respectively. Haplotype analysis further indicated no association of MYOC promoter polymorphisms with the susceptibility for POAG (P > 0.1). On the other hand, there was no difference of POAG phenotypes among different genotypes of MYOC.mt1 (P > 0.31). CONCLUSIONS: In this study on the Chinese population, polymorphisms in the MYOC promoter are not related to the risk of POAG. There is no association between the MYOC.mt1 promoter polymorphism with the severity of POAG. PMID- 15354076 TI - Effect of 5-MCA-NAT, a putative melatonin MT3 receptor agonist, on intraocular pressure in glaucomatous monkey eyes. AB - PURPOSE: 5-MCA-NAT, a putative melatonin MT3 receptor agonist, reduced intraocular pressure (IOP) in ocular normotensive rabbit eyes. This study evaluates the effect of topical application of 5-MCA-NAT on IOP in monkey eyes with laser-induced unilateral glaucoma. METHODS: A multiple-dose study was performed in 8 glaucomatous monkey eyes. One 25-microL drop of 5-MCA-NAT (2%) was applied topically to the glaucomatous eye at 9:30 am and 3:30 pm for 5 consecutive days. IOP was measured hourly for 6 hours beginning at 9:30 am for one baseline day, one vehicle-treated day, and treatment days 1, 3, and 5 with 5 MCA-NAT. RESULTS: Compared with vehicle treatment, twice daily administration of 5-MCA-NAT for 5 days reduced (P < 0.05) IOP from 1 hour to 5 hours after the first dose, and the IOP-lowering effects were shown to last at least 18 hours following administration, based on IOP measurements made after the fourth and eighth doses. The ocular hypotensive effect of 5-MCA-NAT was enhanced with repeated dosing. The maximum reduction (P < 0.001) of IOP occurred at 3 hours after each morning dose, and was 4.0 +/- 0.5 (mean +/- SEM) mm Hg (10%) on day 1, 5.6 +/- 0.8 mm Hg (15%) on day 3, and 7.0 +/- 1.1 mm Hg (19%) on day 5. Adverse ocular or systemic side effects were not observed during the 5 days of treatment. CONCLUSIONS: 5-MCA-NAT, a putative melatonin MT3 receptor agonist, reduces IOP in glaucomatous monkey eyes. Melatonin agonists with activity on the putative MT3 receptor may have clinical potential for treating elevated IOP. PMID- 15354077 TI - Choroidal ganglion cell changes in human glaucomatous eyes. AB - PURPOSE: Choroidal ganglion cells (CGC) are an important source of vasodilative innervation of the posterior ciliary arteries, which also supplies the lamina cribrosa region of the optic nerve. The purpose of this study was to investigate whether CGC undergo quantitative and morphologic changes in human glaucomatous eyes. METHODS: Twenty-five human glaucomatous eyes (15 with primary open angle glaucoma, 10 with pseudoexfoliation glaucoma) and 38 normal eyes (31 age-matched) were investigated. Scleral and choroidal whole mounts were stained for NADPH diaphorase, the CGC quantified in the outer and inner choroid and in the different quadrants separately, and the diameter of the CGC measured. Serial semi and ultrathin sections were studied by light and electron microscopy. RESULTS: In glaucomatous eyes, the total number of CGC was significantly decreased. The most striking loss of CGC was found in the temporal quadrant adjacent to the optic nerve, where only single CGC persisted. The remaining CGC showed a normal ultrastructural appearance but a shift toward larger cells. CONCLUSION: Smaller CGC and those directly adjacent to the optic nerve seem to be most susceptible to the yet unknown pathogenetic factors responsible for this CGC loss. Since the most pronounced loss of CGC occurred in the vicinity of the optic nerve it is tempting to speculate that the vasodilative regulation of the vessels supplying the lamina cribrosa region is impaired in eyes with POAG. PMID- 15354078 TI - Outflow resistance of the Baerveldt glaucoma drainage implant and modifications for early postoperative intraocular pressure control. AB - PURPOSE: To determine outflow resistance of the Baerveldt glaucoma implant using different tube configurations. METHODS: Outflow resistance of 6 tube configurations (C1- C6) of Baerveldt implants was measured under conditions of constant pressure perfusion. Pressures ranged from 2 to 55 mm Hg. Venting slits were created using a 7-0 Vicryl, spatulated suture-needle. Seton tubes were occluded by threading a retrograde suture approximately 1.5 cm into the lumen. RESULTS: At pressures between 2 and 55 mm Hg, mean outflow resistance of the normally configured seton (ie, open tube; C1) was 0.41 (+/- 0.6) mm Hg/microL/min. Resistance was unchanged (mean 0.41 (+/- 0.4) mm Hg/microL/min) by the addition of 4 venting slits (C2) to the seton tube. Occlusion of the open seton tube with a 3-0 Supramid suture (C3) significantly increased (P < 0.001) mean outflow resistance to 14.99 (+/- 0.6) mm Hg/microL/min. Occlusion of the tube with a 4-0 Supramid suture (C4) significantly increased (P < 0.001) mean outflow resistance to 1.09 (+/- 0.5) mm Hg/microL/min. In implants where tubes were occluded with a 3-0 Supramid suture, the addition of venting slits (C5) significantly decreased (P = 0.038) mean outflow resistance to 8.98 (+/- 0.4) mm Hg/microL/min. In tubes occluded with a 4-0 Supramid suture, the addition of venting slits (C6) decreased mean outflow resistance to 0.98 (+/- 0.6) mm Hg/microL/min. CONCLUSIONS: Although these results cannot be directly correlated to the clinical setting, they do show that outflow resistance can be modified at the time of surgery by changing tube configuration of the Baerveldt glaucoma implant. Configuration C5 (3-0 Supramid with venting slits) closely approximates the outflow rate in the normal intraocular pressure range. PMID- 15354079 TI - Flap and suture manipulation after trabeculectomy with adjustable sutures: titration of flow and intraocular pressure in guarded filtration surgery. AB - PURPOSE: To describe the intraocular pressure (IOP) changes resulting from scleral flap suture and scleral flap manipulation in an experimental model of conventional guarded filtration surgery. DESIGN: Experimental study. METHODS: Trabeculectomy operations were performed with adjustable sutures on 5 donor human eyes connected to a constant flow infusion and a real-time IOP monitoring system. Three different manipulations: posterior lip massage, suture adjustment using special forceps, and suture release, were tested on the completed operation site. MAIN OUTCOME MEASURES: The success in lowering IOP to the target range of 7 to 15 mm Hg within 5 interventions, the time to reach equilibrium IOP, and the frequency of undesired outcomes. RESULTS: Posterior lip massage, suture release, and suture adjustment lowered the IOP after stabilization to the target range in less than 5 interventions in 9%, 14%, and 100% respectively (P < 0.001). The median time for IOP to stabilize at the new level after each intervention after correction for the flow rates used was 36.3 minutes for massage, 8.5 minutes for releasable sutures, and 3.7 minutes for adjustment of sutures (P < 0.001). CONCLUSIONS: These results suggest that suture adjustment may be superior to both posterior lip massage and releasable sutures for managing IOP in the early phase following glaucoma surgery. Following clinical interventions that result in loss of anterior chamber volume, IOP checks should be made at least 40 minutes post intervention or at a later time afterwards if there is a clinical risk of low aqueous production. Manipulation of the scleral flap and associated sutures may only lower the IOP for minutes to hours if the suture tension is not decreased. PMID- 15354080 TI - A new model of glaucoma filtering surgery in the rat. AB - PURPOSE: The most common reason for long-term failure of glaucoma filtering surgery (GFS) is scarring of the external filtering "bleb" tissues. The identification of the factors that mediate this process, as well as the development and initial testing of new therapies to limit scarring is enhanced by the use of appropriate animal models. The standard animal model for studying GFS is the rabbit but newer investigative tools that examine changes induced in biologic systems at a genetic level have made development of a rat model desirable. METHODS: Glaucoma filtering surgery was performed on 20 Sprague-Dawley rats by introducing a 30-gauge silicone cannula through a penetrating scleral tunnel, under a limbal-based conjunctival flap and suturing the conjunctiva closed. Identical GFS was performed on 3 additional rats, which underwent histologic evaluation at days 2, 5, and 11, following surgery.Fistulizing surgery was also performed on 6 Sprague-Dawley rats, for comparison, by creating a full thickness needle sclerostomy under a limbal-based conjunctival flap and suturing the conjunctiva closed. RESULTS: Following the cannula GFS, well-elevated filtering blebs formed and these gradually failed over the course of 8 to 13 days. Needle tract sclerostomy filtering blebs formed at the site of the fistulizing surgery but rapidly failed over the course of 2 to 3 days. CONCLUSION: Cannulated filtering surgery in the rat provides a longer lasting and more predictable model than needle tract sclerostomy for studying wound healing following GFS and may facilitate the study of induced changes at the gene level. PMID- 15354081 TI - Reliability of intraocular pressure by Tono-Pen XL over amniotic membrane patch in human. AB - PURPOSE: To determine the reliability of Tono-Pen XL in measuring intraocular pressure (IOP) over the amniotic membrane patch in human eyes. METHOD: Ten healthy volunteers (nine males and one female) participated in this study. After topical anesthesia, IOP was measured using the Tono-Pen XL before and after the application of a single-layer amniotic membrane patch in the right eye and double layer amniotic membrane patch in the left eye. RESULTS: We found no statistical difference between the IOP measured with and without a single layer amniotic membrane (P = 0.808). Nor did we find any statistical difference between the IOP measured with and without double-layer amniotic membrane patch (P = 0.813). The average thickness of the single- and the double-layer amniotic membrane patch measured by pachymetry was 131.9 +/- 31.4 microm (mean +/- SD) and was 246.1 +/- 69.3 microm (mean +/- SD), respectively. CONCLUSIONS: This study showed that it is reliable to measure IOP by Tono-Pen XL over single or a double layer of amniotic membrane patch on healthy human eyes. Further studies are required to determine the accuracy of IOP measurement over amniotic membrane patch on diseased corneas. PMID- 15354082 TI - Glaucoma in Singapore. PMID- 15354083 TI - The aqueous outflow system as a mechanical pump: evidence from examination of tissue and aqueous movement in human and non-human primates. AB - PURPOSE: To describe a new aqueous outflow model involving a mechanical pump. MATERIALS AND METHODS: Laboratory materials include human and monkey eyes; methods include the dissecting microscope, light microscopy, scanning electron microscopy, transmission electron microscopy, and tracer studies. Clinical methods involve human subject slit lamp, gonioscopy, and operating microscope examination. RESULTS: Laboratory evidence demonstrates that aqueous outflow tissues are responsive to intraocular pressure induced deformation. Deformation occurs in response to small pressure gradients. Laboratory evidence also demonstrates the presence of valves discharging aqueous to Schlemm's canal. The laboratory model predicts pulsatile aqueous discharge in vivo. Clinical in vivo evidence demonstrates pulsatile aqueous flow from the anterior chamber into Schlemm's canal, from Schlemm's canal into collector channels, and from Schlemm's canal into aqueous and episcleral veins, all synchronous with the ocular pulse. CONCLUSIONS: Aqueous outflow tissue deformation caused by normal intraocular pressure transients induces pulsatile one-way discharge of aqueous to the vascular system. The model identifies biomechanical coupling of intraocular pressure with aqueous outflow tissue deformation and also sites of high flow capable of inducing shear stress. These mechanotransduction mechanisms, well characterized as a means of controlling pressure and flow in the vascular system, also provide a means of regulatory feedback to control intraocular pressure and aqueous flow. PMID- 15354084 TI - Alterations in expression of angiopoietins and the Tie-2 receptor in the retina of streptozotocin induced diabetic rats. AB - PURPOSE: The angiopoietin (Ang)/Tie-2 system may play a role in vascular integrity and angiogenesis. In this study, we investigated alterations of the gene expression of Ang-1 and Ang-2 in the retinas of streptozotocin (STZ) induced diabetic rats. METHODS: In situ hybridization, reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analyses were performed to determine the mRNA and protein content for Ang-1 and Ang-2 and the Tie2 receptor in the retinas of STZ diabetic and age matched control rats. RESULTS: Using in situ hybridization analysis, Ang-1, Ang-2, and Tie2 mRNA expression was observed in the ganglion cell layer (GCL) and the inner nuclear layer (INL). While Ang-2 mRNA expression did not changed after 2 weeks, 1 month, or 3 months of STZ induced diabetes, it was increased in the GCL and slightly elevated in the INL after 6 months of diabetes. In contrast, Ang-1 and Tie2 mRNA expression was stable at every timepoint during 6 months of STZ induced diabetes. RT-PCR and western blot analyses confirmed the increase of Ang-2 expression after 6 months of diabetes. Furthermore, double staining of alpha-smooth muscle actin (alphaSMA) and Ang-2 mRNA demonstrated that the SMA positive cells surrounding Ang-2-expressing cells were decreased in the GCL. CONCLUSIONS: Diabetes increases Ang-2 expression in the GCL accompanied by a reduction of alphaSMA positive perivascular cells. These changes may suggest a role for Ang-2 in the mechanism of pericyte loss in diabetic retinopathy. PMID- 15354085 TI - Immunolocalization and regulation of iron handling proteins ferritin and ferroportin in the retina. AB - PURPOSE: CNS iron accumulation is associated with several neurodegenerative diseases, including age-related macular degeneration. Intracellular overload of free iron is prevented, in part, by the iron export protein, ferroportin, and the iron storage protein, ferritin. The purpose of this study was to assess retinal localization and regulation of ferroportin and ferritin. METHODS: Normal murine retinas were analyzed by immunohistochemistry to localize ferroportin, cytosolic ferritin, and mitochondrial ferritin, with double-labeling using cell-specific markers to identify cell types. Retinas deficient in the ferroxidases, ceruloplasmin and hephaestin, accumulate iron in their retinas and RPE, while retinas deficient in iron regulatory proteins (IRPs) lack the ability to regulate several proteins involved in iron metabolism; retinas from these knockout mice along with their age matched wild type littermates were also examined to study regulation of ferritin and ferroportin. To enable visualization of label in the retinal pigment epithelial cells, sections from pigmented mice were bleached with H2O2 prior to IHC, a novel use of this technique for study of the RPE. RESULTS: In normal retinas, cytosolic ferritins were found predominantly in rod bipolar cells and photoreceptors. Ferroportin was found in RPE and Muller cells. Iron accumulation in mice deficient in ceruloplasmin and hephaestin was associated with upregulation of ferritin and ferroportin. Mice deficient in IRPs showed upregulation of ferritin and ferroportin, likely because of their inability to repress translation. CONCLUSIONS: Normal retinas contain ferritin and ferroportin, whose levels are regulated by iron-responsive, iron regulatory proteins. Ferroportin colocalizes with ceruloplasmin and hephaestin to RPE and Muller cells, supporting a potential cooperation between these ferroxidases and the iron exporter. Cytosolic ferritin accumulates in rod bipolar synaptic terminals, suggesting that ferritin may be involved in axonal iron transport. Mitochondrial ferritin increases with iron accumulation, suggesting a role in iron storage. PMID- 15354087 TI - Caveat emptor: the treachery of work-up bias. PMID- 15354089 TI - Gender differences in lung cancer: have we really come a long way, baby? PMID- 15354090 TI - Gender and lung transplantation: size matters, does sex? PMID- 15354091 TI - Effect of carbon dioxide insufflation on free internal thoracic artery flows: is it a vasodilator? AB - BACKGROUND: This study was conceived to evaluate the effect of carbon dioxide insufflation on free internal thoracic artery flows. METHODS: We studied 56 consecutive patients who underwent coronary artery bypass grafting in which the left internal thoracic artery was anastomosed to the left anterior descending artery. The first 26 consecutive internal thoracic arteries were harvested as a pedicled graft (group 1), and the next 30 consecutive internal thoracic arteries were dissected by using the carbon dioxide insufflation technique (group 2). The internal thoracic artery harvesting was performed by 2 experienced surgeons by using the same instrumentation and technique. First, free flows of the internal thoracic arteries were registered after distal cutting of the vessel in both groups. After the first measurements, diluted papaverine was sprayed on the internal thoracic artery pedicle only in group 1, and then second measurements were registered after 15 minutes in both groups. Hemodynamic parameters were recorded with each measurement. RESULTS: The first free flow measurement was significantly higher in the carbon dioxide-insufflated internal thoracic arteries (group 2, 60 +/- 32 mL/min; group 1, 28 +/- 19 mL/min; P <.05). Although the second free flow measurement of the carbon dioxide-insufflated group was higher than in group 1, the difference was not statistically significant (68 +/- 46 mL/min vs 53 +/- 32 mL/min; P =.53). CONCLUSIONS: Carbon dioxide insufflation of the internal thoracic artery is an efficient technique to increase the flow and seems to be safe, simple, and reliable. When the internal thoracic artery is harvested in a carbon dioxide-insufflated fashion, arterial spasm and reduced early flow may be avoided, even without vasodilator agents such as papaverine. PMID- 15354092 TI - Locally applied cilostazol suppresses neointimal hyperplasia by inhibiting tenascin-C synthesis and smooth muscle cell proliferation in free artery grafts. AB - OBJECTIVE: Accumulation of smooth muscle cells and extracellular matrix in the intima of artery bypass grafts induces neointimal hyperplasia, resulting in graft failure. We investigated the inhibitory effect of locally applied cilostazol, an inhibitor of cyclic adenosine monophosphate phosphodiesterase III, on neointimal hyperplasia and the role of tenascin-C synthesis and smooth muscle cell proliferation in free artery grafts. Methods and results We established a distal anastomotic stricture model of free artery graft stenosis using rat abdominal aorta. In this model, neointimal hyperplasia was observed not only in the distal anastomotic site but also in the graft body at postoperative day 14 and was markedly progressed at day 28. Strong expression of tenascin-C was found in the media and neointima of the graft body. When cilostazol was locally administered around the graft using Pluronic gel, neointimal hyperplasia of the graft was significantly suppressed in comparison with gel-treated control graft. The mean neointima/media area ratio was reduced by 86.6% for the graft body and by 75.8% for the distal anastomotic site versus the control. Cilostazol treatment decreased cell proliferation and tenascin-C expression in the neointima. In an in vitro experiment using cultured smooth muscle cells isolated from rat aorta, cilostazol completely suppressed the tenascin-C mRNA expression induced by platelet-derived growth factor-BB. CONCLUSION: A single topical administration of cilostazol may suppress neointimal hyperplasia by inhibiting cell proliferation and tenascin-C synthesis in free artery grafts, presenting the potential for clinical use in vascular surgery. PMID- 15354093 TI - Warm blood cardioplegic arrest induces mitochondrial-mediated cardiomyocyte apoptosis associated with increased urocortin expression in viable cells. AB - OBJECTIVES: This study assesses the mechanisms of apoptosis in patients after on pump coronary artery bypass graft surgery and the potential involvement of the endogenous cardiac peptide urocortin as a cardiomyocyte salvage mechanism. We have previously described the mechanisms of apoptosis after ischemia-reperfusion injury in the rat heart and shown that endogenous urocortin is cardioprotective. Here we extend these findings to the human heart exposed to ischemic-reperfusion injury. METHODS: Two sequential biopsy specimens were obtained from the right atriums of 24 patients undergoing coronary artery bypass grafting at the start of grafting and 10 minutes after release of the aortic clamp. Apoptosis was identified by means of immunocytochemical colocalization between terminal deoxynucleotidyl transferase-mediated nick end-labeling positivity and active caspase-3. Immunostaining for active caspase-9 and caspase-8 was performed to identify the pathways of apoptosis induction. Urocortin and adenosine triphosphate-dependent potassium channel expression was also assessed by means of immunocytochemistry. RESULTS: Myocyte apoptosis (<0.1% before coronary artery bypass grafting) was increased after coronary artery bypass grafting and reperfusion and was greater in patients with longer periods of cardioplegic arrest (3.3% +/- 0.5% with <55 minutes and 5.1% +/- 0.9% with 85-100 minutes, P <.001). Processing of caspase-9 was always more pronounced than that of caspase-8 (P <.05). Cardioplegic arrest was also associated with increased urocortin expression (up to 29% +/- 3.5% vs <3% in samples obtained before coronary artery bypass grafting, P <.001) but only in nonapoptotic myocytes. These and surrounding viable myocytes also showed increased Kir6.1 adenosine triphosphate dependent potassium channel expression. CONCLUSIONS: Cardioplegic arrest and subsequent reperfusion result in cardiomyocyte apoptosis, largely through mitochondrial injury, as well as exclusive urocortin expression in viable cells. This finding might suggest a cardioprotective role for endogenous urocortin in human subjects. PMID- 15354094 TI - Development of an artificial vessel lined with human vascular cells. AB - OBJECTIVES: Thrombogenity of small-diameter vascular prostheses might be reduced by complete coverage of the luminal surface with vascular cells. We investigated cell seeding on polyurethane vascular prostheses. METHODS: Thirty polyurethane vascular prostheses were divided into 3 groups of 10 each: group A, diameter of 20 mm and gamma-sterilized; group B, diameter of 4 mm and gamma-sterilized; and group C, diameter of 4 mm and ethylene oxide sterilized. Human smooth muscle cells, fibroblasts, and endothelial cells were isolated from saphenous vein segments and expanded in culture. Five polyurethane vascular prostheses of each group were seeded with endothelial cells alone (mean, 4.8 +/- 1.2 x 10(6) cells), and the remaining 5 polyurethane vascular prostheses were preseeded with a mixed culture of fibroblasts and smooth muscle cells (mean, 7.7 +/- 2.3 x 10(6) cells), followed by endothelial cell seeding (mean, 4.4 +/- 0.9 x 10(6) cells). Seven days after cell seeding, the polyurethane vascular prostheses were perfused under a pulsatile flow (80 pulses/min, 140/80 mm Hg, and 120 mL/min) for 2 hours. Specimens were taken after each seeding procedure both before and after perfusion and then examined both with a scanning electron microscope and immunohistochemically. RESULTS: Isolated endothelial cell seeding revealed better initial adhesion in groups A and B than in group C (63% vs 33%). After 7 days, the cells had covered approximately 80% of the luminal surface in groups A and B, whereas group C cells rounded up and lost adhesion. After perfusion testing of group A and B prostheses, only 10% of the surface was still covered with endothelial cells. Preseeding with the mixed culture again revealed a better initial adhesion in groups A and B compared with that in group C (76% vs 41%). In groups A and B endothelial cell seeding (adhesion, 72%) resulted in a confluent endothelial cell layer. The results of immunohistochemical staining were positive for collagen IV, laminin, CD31, and Factor VIII. In group C only isolated cells were found after each seeding procedure, which rounded up and vanished during the next days. Perfusion testing of group A and B prostheses revealed that the confluent cell layer remained stable, with only small defects (<10% of the surface). The cells stained positivively for endothelial nitric oxide synthase. CONCLUSION: Seeding of a mixed culture out of fibroblasts and smooth muscle cells resulted in improved endothelial cell adhesion and resistance to shear stress. This outcome was caused by an increased synthesis of extracellular matrix proteins. Cell attachment was better on gamma-sterilized polyurethane vascular prostheses compared with on those undergoing ethylene oxide sterilization. PMID- 15354095 TI - Comparison of pH management during antegrade selective cerebral perfusion in canine models with old cerebral infarction. AB - OBJECTIVE: We sought to examine the influence on the brain, with or without old infarction, of pH management during antegrade selective cerebral perfusion in a canine model. METHODS: A cerebral infarct canine model was created by injecting a cylindrical silicone embolus. Dogs that had obvious neurologic deficits and had survived for 4 weeks or more were included in the model. Deep hypothermia with antegrade selective cerebral perfusion was performed in intact mongrel dogs (alpha-stat: group A, n = 6; pH-stat: group B, n = 6) and mongrel dogs with infarctions (alpha-stat: group C, n = 6; pH-stat: group D, n = 6). Maxillary vein saturation of oxygen, venous-arterial lactate difference, and serum concentrations of malondialdehyde and glutamate were measured and central conduction times and amplitude in somatosensory evoked potentials were assessed during the operation. RESULTS: During the experimental procedure, the maxillary vein saturation of oxygen was significantly less (P <.05), whereas the venous arterial lactate difference was significantly greater (P <.05) in the cooling phase to 28 degrees C in group C than in the other groups. The pH-stat group showed significantly greater arterial Paco(2) and lower pH than the alpha-stat group during the period between the cooling to 28 degrees C and the rewarming to 28 degrees C (P <.05). Other intraoperative parameters did not show any difference among the groups. In group C the serum concentrations of malondialdehyde and glutamate significantly increased, as did the central conduction time, whereas in both groups C and D the amplitude ratio decreased significantly. CONCLUSIONS: This experiment suggests that pH-stat management during antegrade selective cerebral perfusion provides more effective protection for a brain with old infarction than alpha-stat management. PMID- 15354096 TI - Axillary artery cannulation for cardiopulmonary bypass reduces cerebral microemboli. AB - BACKGROUND: Aortic cannulation for cardiopulmonary bypass (CPB) is linked to cerebral microemboli emanating from the ascending aorta. Aortic calcification or disease requiring replacement precludes aortic cannulation. Clinical experience with axillary artery cannulation led to the hypothesis that axillary cannulation may be cerebroprotective. METHODS: Five mongrel dogs underwent a median sternotomy and isolation of the right axillary artery. The canine bicarotid brachiocephalic trunk was reconfigured by grafting the origin of the left carotid to the proximal left subclavian artery. Microspheres were injected into the ascending aorta during 4 conditions: before and after reconfiguration, CPB with aortic cannulation, and CPB with axillary cannulation. Brain, kidneys, and skeletal muscle were analyzed for microsphere distribution. RESULTS: Each animal served as its own control for comparison of aortic and axillary cannulation. No significant differences were documented in microsphere deposition for prereconfiguration and postreconfiguration. In the right middle cerebral artery distribution, 2300 +/- 710 microspheres per gram were deposited during aortic cannulation, compared with 540 +/- 110 during axillary cannulation (P <.05). In the left middle cerebral artery region, 2030 +/- 330 microspheres per gram with aortic cannulation were reduced to 1320 +/- 240 with axillary cannulation (P <.05). Axillary cannulation resulted in 73% fewer microspheres in the right brain and 40% fewer microspheres in the left compared with aortic cannulation (P <.05). CONCLUSIONS: Axillary artery cannulation for CPB is cerebroprotective. Altered blood-flow patterns during axillary cannulation may produce retrograde brachiocephalic artery blood flow and competing intracerebral right-to-left collateral blood flow, deflecting emboli from the ascending aorta and arch toward the descending aorta. Expanded use of axillary artery cannulation during cardiac operations could decrease the incidence of stroke. PMID- 15354097 TI - Improved myocardial function after transmyocardial laser revascularization according to cine magnetic resonance in a porcine model. AB - OBJECTIVE: This study was undertaken to demonstrate that transmyocardial laser revascularization of hypoperfused myocardium improves regional and global myocardial function. METHODS: Cine magnetic resonance imaging was used to monitor regional wall thickening (in millimeters) and cardiac output (in milliliters per kilogram per minute). Cine magnetic resonance imaging was performed before and 8 weeks after transmyocardial laser revascularization was applied to the hypoperfused lateral wall of the left ventricle (target area) in a porcine model (n = 9, transmyocardial laser revascularization group). A second group of animals was left untreated (n = 8, control group). RESULTS: Regional wall thickening in the target area improved after transmyocardial laser revascularization (0.7 +/- 0.3 mm to 3.7 +/- 1.9 mm, P <.02) and was significantly higher (P <.01) after transmyocardial laser revascularization than in the control group, in which it did not improve (0.5 +/- 0.6 mm to 0.5 +/- 1.2 mm). Accordingly, cardiac output and microsphere-derived myocardial blood flows were significantly higher than in the control group (P <.01), and the amount of triphenyltetrazolium chloride stained myocardium was lower (P <.01). CONCLUSION: Cine magnetic resonance imaging demonstrates improved global and regional myocardial function after transmyocardial laser revascularization in a porcine model. PMID- 15354098 TI - Fluorine 18-tagged fluorodeoxyglucose positron emission tomographic scanning to predict lymph node metastasis, invasiveness, or both, in clinical T1 N0 M0 lung adenocarcinoma. AB - OBJECTIVE: We sought to predict lymph node metastasis and tumor invasiveness in clinical T1 N0 M0 lung adenocarcinomas, and we measured fluorodeoxyglucose uptake on positron emission tomography. METHODS: Fluorodeoxyglucose positron emission tomography was performed on 44 patients with adenocarcinomas of 1 to 3 cm in size clinically staged as T1 N0 M0 before major lung resection with lymph node dissection. Fluorodeoxyglucose uptake was evaluated by using the contrast ratio between the tumor and contralateral healthy lung tissue. Lymphatic and vascular invasion within tumors, pleural involvement, and grade of histologic differentiation were examined. RESULTS: The pathologic tumor stage was T1 N0 M0 in 36 patients, and a more advanced stage was found in 8 patients. Although all 22 adenocarcinomas with a contrast ratio of less than 0.5 in fluorodeoxyglucose uptake were pathologic T1 N0 M0 tumors, 8 (36%) of 22 with a contrast ratio of 0.5 or greater were of a more advanced stage than T1 N0 M0, with the difference being significant (P =.002). Adenocarcinomas with a contrast ratio of less than 0.5 showed less lymphatic and vascular invasion and less pleural involvement than those with a contrast ratio of 0.5 or greater (P =.006, P =.004, and P =.02, respectively). The grade of histologic differentiation was well differentiated in 19 of 22 adenocarcinomas with a contrast ratio of less than 0.5 (86%), which was a greater frequency than the 4 (18%) of 22 adenocarcinomas with a contrast ratio of 0.5 or greater (P <.001). CONCLUSION: Clinical T1 N0 M0 lung adenocarcinomas with a contrast ratio of less than 0.5 usually did not have lymph node metastasis, had less tumor involvement of vessels or pleura, and were more frequently well differentiated than those with a contrast ratio of 0.5 or greater. Limited lung resection could be indicated, lymph node dissection or mediastinoscopy could be reduced, or both in this type of adenocarcinoma. PMID- 15354099 TI - Long-term outcome of Heller myotomy in achalasic sigmoid esophagus. AB - OBJECTIVE: We sought to assess the long-term outcome of Heller myotomy and anterior fundoplication in patients with achalasic sigmoid esophagus. METHODS: Fourteen patients with achalasia and sigmoid esophagus (median age, 42.5 years) operated on by the same surgeon through a laparotomy (n = 8) or laparoscopic approach (n = 6) between 1985 and 2000 were evaluated. According to a 4-grade classification (1, no symptoms; 4, persistent symptoms), both dysphagia and regurgitation had a median score of 4.0. Five patients complained of respiratory symptoms. Six patients had undergone previous pneumatic dilation. Preoperative and postoperative workup included an esophagogram, esophagoscopy, manometry, and health-related quality-of-life assessment with the Short-Form 36-item questionnaire. RESULTS: Median follow-up was 85 months. At 24 months, esophageal width decreased by 10 mm (P =.003), and the change correlated inversely with the age of the patients (R = -0.61; P =.02). Lower esophageal sphincter pressure decreased by 17 mm Hg (P =.001), and both dysphagia and regurgitation scores decreased to 1.0 (P <.003). Comparison with the results of 37 patients with earlier-stage achalasia showed no difference in changes of esophageal width, lower esophageal sphincter pressure, dysphagia score, and regurgitation score. Quality-of-life Short-Form 36-item questionnaire domains, including general health, social functioning, and vitality, improved significantly. Overall results were classified as excellent or good in 10 patients and as satisfactory and unsatisfactory in 2 patients each. No patient required esophagectomy or had esophageal carcinoma. CONCLUSIONS: In this study Heller myotomy proved effective in improving subjective, objective, and quality-of-life outcome measures in patients with achalasic sigmoid esophagus and should be considered as the first choice treatment for this severe condition. PMID- 15354100 TI - Long-term results after lung volume reduction surgery in patients with alpha1 antitrypsin deficiency. AB - BACKGROUND: The favorable effects of lung volume reduction surgery for selected patients with smoker's emphysema has been demonstrated. However, outcome data for patients with alpha(1)-antitrypsin deficiency emphysema are scarce. METHODS: We prospectively studied pulmonary function, dyspnea, and 6-minute walking distance in 21 patients with severe alpha(1)-antitrypsin deficiency emphysema (PiZZ 18, PiZO 1, PiSZ 2, 10 female patients, median age 56 years, range 38-74 years) for as long as 5 years after thoracoscopic lung volume reduction surgery. RESULTS: Lung volume reduction surgery improved the mean dyspnea score, from 3.7 +/- 0.1 preoperatively to 1.4 +/- 0.2 at 3 months; the score remained improved for as long as 3.5 years. Mean vital capacity (% predicted) improved from 79% +/- 4.4% to 98% +/- 4.8% at 3 months, and the ratio of residual volume to total lung capacity decreased from 0.67 to 0.51. These improvements lasted for as long as 2 years. The mean airflow obstruction (forced expiratory volume in 1 second % predicted) improved from 27% +/- 1.9% to 38% +/- 3.3% at 3 months and remained statistically improved for 1 year. Four patients showed long-term improvement in lung function for as long as 3.5 years. These patients had markedly heterogeneous emphysema and showed no radiologic signs of airway inflammation. CONCLUSIONS: Lung volume reduction surgery in patients with advanced emphysema from alpha(1) antitrypsin deficiency results in a significant improvement in dyspnea and lung function for as long as 3.5 years in some cases. It appears that magnitude and duration of these effects are inferior and shorter than those in patients with pure smoker's emphysema. Patients with heterogeneous disease and no or minor inflammatory airway disease may benefit most. PMID- 15354101 TI - Postoperative chemotherapy for non-small cell lung cancer: A systematic review and meta-analysis. AB - BACKGROUND: Postoperative chemotherapy is currently not recommended for resected non-small cell lung cancer in many countries and centers. Recently, results of several large randomized clinical trials were reported with conflicting evidence. Accordingly, we sought to determine whether postoperative chemotherapy is associated with improved survival compared with that after surgical intervention alone. METHODS: Randomized clinical trials with cisplatin- or uracil plus ftorafur-containing regimens were included and evaluated separately. A systematic review that included randomized clinical trials performed before 1995 was identified and found to be of adequate quality. Further randomized controlled trials were identified by searching MEDLINE, EMBASE, and the Cochrane Controlled Trials Register from 1995 through 2004. In addition, the reference lists of articles and conference abstracts were searched. The logarithm of the hazard ratio and its standard error were calculated, and a fixed-effect model was used to combine the estimates. RESULTS: There were 7200 patients enrolled in 19 trials included in the analyses. An overall estimate of 13% relative reduction in mortality (95% confidence interval, 7%-19%) was found. There was 11% relative reduction in mortality associated with postoperative cisplatin (95% confidence interval, 4%-18%; P =.004) and 17% associated with uracil plus ftorafur (95% confidence interval, 5%-27%; P =.006) compared with that after surgical intervention alone. This means that there would be an additional survivor at 5 years for 25 patients treated with cisplatin or for 30 patients treated with uracil plus ftorafur. CONCLUSIONS: Postoperative chemotherapy is associated with improved survival compared with that after surgical intervention alone. Selected patients with completely resected non-small cell lung cancer should be offered chemotherapy. PMID- 15354102 TI - Sleeve segmentectomy for non-small cell lung carcinoma. AB - OBJECTIVE: Although sleeve segmentectomy for centrally located lung cancers was originally designed for patients unable to tolerate lobectomy, we have tried it in patients with noncompromised function as well. We evaluated the efficiency of this atypical type of bronchoplasty. METHODS: Of 202 patients for whom we performed bronchoplasty for primary non-small cell lung carcinoma, 16 underwent sleeve segmentectomy. RESULTS: Sixteen patients were classified into 4 groups according to the mode of bronchial reconstruction: type A, anastomosis between the right intermediate or left main and basal segmental bronchi with removal of the superior segment of the lower lobe (S6; n = 7); type B, anastomosis between the left main and lingular bronchi with removal of the upper division of the left upper lobe (S1+2+3; n = 3); type C, anastomosis between the left main and upper division bronchi with removal of the lingular segments (S4+5; n = 4); and type D, others (n = 2). Nine patients had pulmonary function sufficient to tolerate lobectomy. The tumors were completely resected in all patients. Combined performance of pulmonary angioplasty was carried out in 2 patients. Bronchial reconstruction was successful in all patients, with neither bronchial complications nor local recurrences. Ten patients had stage IA disease, and 6 had more advanced disease. All patients were alive, except 1 who died as a result of distant metastasis and 2 who died of noncancerous causes. Overall 3-year and 5 year survivals were 93.3% and 68.1%, respectively. CONCLUSIONS: Sleeve segmentectomy, which is technically demanding, should be considered in patients with centrally located and possibly curable early non-small cell lung cancer because the prevalence of small-sized or multiple lung tumors has been increasing and because our findings suggest that this lung-saving operation is safe and useful. PMID- 15354103 TI - Clopidogrel and bleeding in patients undergoing elective coronary artery bypass grafting. AB - OBJECTIVE: In an effort to minimize transfusions in patients undergoing elective coronary artery bypass grafting operations after recent clopidogrel exposure, we studied laboratory tests predictive of platelet dysfunction and used a strict algorithm-driven treatment of bleeding. METHODS: Forty-five patients receiving clopidogrel within 6 days of the operation and 45 control subjects were studied. Prothrombin time, activated partial thromboplastin time, platelet count, and platelet function test results were measured before heparinization, after protamine administration, and then every 2 hours. No transfusions were administered unless a patient met both laboratory and clinical criteria. RESULTS: Algorithm-driven treatment of bleeding significantly reduced the mean units of all blood components transfused by about one third, as shown by comparison with current control and historical data. Compared with current control subjects, clopidogrel recipients required significantly more transfusions of platelets (9.0 +/- 1.7 vs 1.2 +/- 0.5 U; P <.0001) and packed red blood cells (4.3 +/- 0.6 vs 2.3 +/- 0.5 U; P =.01) and required longer periods of controlled ventilation (12.4 +/- 1.3 vs 8.6 +/- 0.8 hours; P =.02). Preoperative platelet dysfunction before heparin administration for cardiopulmonary bypass, as measured by using adenosine diphosphate aggregometry (response <40%), predicted all but 1 case of severe coagulopathy requiring multiple transfusions (16.6 +/- 2.8 U of platelets and 5.8 +/- 1.0 U of packed red blood cells). CONCLUSIONS: A strict transfusion algorithm can reduce the transfusion requirement for all blood components. Preheparin testing of platelet function with adenosine diphosphate aggregometry can identify patients at highest risk for perioperative bleeding and transfusions and might further reduce the perioperative transfusion requirement. PMID- 15354104 TI - Clopidogrel did not inhibit platelet function early after coronary bypass surgery: A prospective randomized trial. AB - OBJECTIVE: Although the beneficial effect of aspirin prescription after coronary surgery has been established, the efficacy of clopidogrel has never been compared with that of aspirin in the critical early postoperative period. We therefore conducted a prospective, double-blind, randomized controlled trial to compare the efficacies of these antiplatelet regimens. METHODS: Patients undergoing elective primary coronary artery bypass surgery were invited to participate. After the operation, patients were randomized to receive 100 mg aspirin, 325 mg aspirin, or 75 mg clopidogrel tablets daily for 5 days. Our primary outcome measure was platelet aggregation on day 5, expressed as percentage of baseline. Assessment of platelet aggregation was undertaken with the technique of Born. RESULTS: From September 2002 to July 2003, a total of 54 patients were randomized into the study. There were 2 self-withdrawals and 2 protocol violations, leaving 50 patients for analysis, 34 in the aspirin group and 16 in the clopidogrel arm. Compared with baseline, the mean percentage aggregations with collagen on day 5 were 56% for aspirin and 99% for clopidogrel. The mean difference between the two arms was 42% (95% confidence interval 27%-56%) in favor of aspirin. At the same time point, the effective concentration to inhibit 50% aggregation in the samples from patients randomly assigned to receive clopidogrel were not raised for our entire panel of agonists (changes of -0.04 microg/L for collagen, -0.01 micromol/L for epinephrine, and -0.02 micromol/L for adenosine diphosphate). CONCLUSION: Clopidogrel, unlike aspirin, did not inhibit platelet aggregation in the first 5 postoperative days and therefore should not be used as a sole antiplatelet agent early after coronary surgery. PMID- 15354105 TI - Aortic valve replacement with the minimal extracorporeal circulation (Jostra MECC System) versus standard cardiopulmonary bypass: a randomized prospective trial. AB - BACKGROUND: We prospectively evaluated a newly introduced minimal extracorporeal circulation system (Jostra MECC System; Jostra AG, Hirrlingen, Germany) for aortic valve surgery. METHOD: In a prospective, randomized study, 100 patients underwent aortic valve replacement either with standard cardiopulmonary bypass (n = 50, group B) or with the MECC System (n = 50, group B). The myocardial protection and the left vent were identical for the two groups. The intrapericardial suction device was never used (only the cell salvage device was used) to reduce the air-blood contact area. RESULTS: No significant differences were noted in patient characteristics and operative data between groups. Operative mortality (<30 days) was 2% for group A and 4% for group B (difference not significant). From the preoperative period to the postoperative period, the increase in C-reactive protein was significantly higher for group B (P <.001). The postoperative troponin I level was significantly lower in group A (mean 4.65 +/- 2.9 microg/L at 24 hours) than in group B (8.2 +/- 4.4 microg/L, P <.03). On the other hand, the MECC System was associated with platelet preservation. Renal function was better preserved and the neurologic event rate was significantly lower for the MECC group (P <.02). CONCLUSION: The MECC System is safe and allows aortic valve replacement under the most favorable conditions. The system is more biocompatible than standard cardiopulmonary bypass and provides a good postoperative biologic profile and good clinical results, particularly for high risk patients. PMID- 15354106 TI - Effect of aprotinin on clinical outcomes in coronary artery bypass graft surgery: a systematic review and meta-analysis of randomized clinical trials. AB - OBJECTIVE: Despite proven blood transfusion benefits, aprotinin may be underused in coronary artery bypass grafting. Reluctance to use aprotinin may stem from safety concerns. The current objective was to evaluate clinical outcomes (mortality, myocardial infarction, renal failure, stroke, atrial fibrillation) in patients undergoing coronary artery bypass grafting who receive aprotinin by performing a quantitative overview of published, randomized, controlled trials. METHODS: MEDLINE, EMBASE, and PHARMLINE (1988-2001) and reference lists of relevant articles were searched for coronary artery bypass grafting studies. Criteria for data inclusion were as follows: (1) random allocation of study treatments, (2) placebo control, (3) enrollment only of patients undergoing coronary artery bypass grafting, (4) no combination with another experimental medication or device, and (5) prophylactic and continuous intraoperative use. RESULTS: Data from 35 coronary artery bypass grafting trials (n = 3879) confirm that aprotinin reduces transfusion requirements (relative risk 0.61, 95% confidence interval 0.58-0.66) relative to placebo, with a 39% risk reduction. Aprotinin therapy was not associated with increased or decreased mortality (relative risk 0.96, 95% confidence interval 0.65-1.40), myocardial infarction (relative risk 0.85, 95% confidence interval 0.63-1.14), or renal failure (relative risk 1.01, 95% confidence interval 0.55-1.83) risk, but it was associated with a reduced risk of stroke (relative risk 0.53, 95% confidence interval 0.31-0.90) and a trend toward reduced atrial fibrillation (relative risk 0.90, 95% confidence interval 0.78-1.03). CONCLUSIONS: Aprotinin reduces transfusion requirements. Concerns that aprotinin therapy is associated with increased mortality, myocardial infarction, or renal failure risk is not supported by data from published, randomized, placebo-controlled clinical trials. Evidence for a reduced risk of stroke and a tendency toward reduction of atrial fibrillation occurrence was observed in patients who received aprotinin. PMID- 15354107 TI - Surgery for left ventricular aneurysm: early and late survival after simple linear repair and endoventricular patch plasty. AB - OBJECTIVE: Simple linear resection and endoventricular patch plasty are alternative techniques to repair postinfarction left ventricular aneurysm. The aim of the study was to compare these 2 methods with regard to early mortality and long-term survival. METHODS: We retrospectively reviewed 159 patients undergoing operations between 1989 and 2003. The epidemiologic design was of an exposed (simple linear repair, n = 74) versus nonexposed (endoventricular patch plasty, n = 85) cohort with 2 endpoints: early mortality and long-term survival. The crude effect of aneurysm repair technique versus endpoint was estimated by odds ratio, rate ratio, or relative risk and their 95% confidence intervals. Stratification analysis by using the Mantel-Haenszel method was done to quantify confounders and pinpoint effect modifiers. Adjustment for multiconfounders was performed by using logistic regression and Cox regression analysis. Survival curves were analyzed with the Breslow test and the log-rank test. RESULTS: Early mortality was 8.2% for all patients, 13.5% after linear repair and 3.5% after endoventricular patch plasty. When adjusted for multiconfounders, the risk of early mortality was significantly higher after simple linear repair than after endoventricular patch plasty (odds ratio, 4.4; 95% confidence interval, 1.1 17.8). Mean follow-up was 5.8 +/- 3.8 years (range, 0-14.0 years). Overall 5-year cumulative survival was 78%, 70.1% after linear repair and 91.4% after endoventricular patch plasty. The risk of total mortality was significantly higher after linear repair than after endoventricular patch plasty when controlled for multiconfounders (relative risk, 4.5; 95% confidence interval, 2.0 9.7). Linear repair dominated early in the series and patch plasty dominated later, giving a possible learning-curve bias in favor of patch plasty that could not be adjusted for in the regression analysis. CONCLUSIONS: Postinfarction left ventricular aneurysm can be repaired with satisfactory early and late results. Surgical risk was lower and long-term survival was higher after endoventricular patch plasty than simple linear repair. Differences in outcome should be interpreted with care because of the retrospective study design and the chronology of the 2 repair methods. PMID- 15354108 TI - Small aortic annulus: the hydrodynamic performances of 5 commercially available bileaflet mechanical valves. AB - OBJECTIVE: Hemodynamic performances of mechanical valve prostheses in patients with aortic valve stenosis and a small aortic annulus are crucial. We analyzed the in vitro hydrodynamics of 5 currently available bileaflet mechanical prostheses that fitted a 21-mm-diameter valve holder of a Sheffield pulse duplicator. METHODS: Three samples of 5 high-performance production-quality prostheses, including the sewing ring cuffs, were tested in the aortic chamber of a Sheffield pulse duplicator. Sizes of the prostheses fitting the 21-mm valve holder were as follows: 18-mm ATS, 19-mm SJM Regent, 19-mm Sorin Bicarbon Slimline, 19-mm On-X, and 21-mm Carbomedics Top Hat. The tests were carried out at a fixed pulse rate (70 beats/min) and at increasing cardiac outputs of 2, 4, 5, and 7 L/min. Each valve was tested 10 times for each different cardiac output. This resulted in a total of 40 tests for each valve and 120 tests for each valve model. The aortic pressure was set at 120/80 mm Hg (mean pressure, 100 mm Hg) throughout the experiment for all cardiac outputs. Forward flow pressure decrease, total regurgitant volume, closing and leakage volumes, effective orifice area, and stroke work loss were recorded while the valve operated under each cardiac output. RESULTS: The SJM Regent valve and the Sorin Bicarbon Slimline prosthesis showed the lowest mean and peak gradients at increasing cardiac outputs. The closure volume was higher for the SJM Regent and Sorin Bicarbon Slimline prostheses, unlike with the ATS prosthesis at 7 L/min of cardiac output. The ATS and SJM Regent prostheses showed the largest regurgitant volume, whereas the Sorin Bicarbon Slimline prosthesis showed the lowest regurgitant volume. The calculated effective orifice area and stroke work loss were significantly better with the SJM Regent and Sorin Bicarbon Slimline prostheses. CONCLUSION: Assuming that the 21-mm valve holder in which all the tested prostheses were accommodated is comparable with a defined aortic valve size, this hydrodynamic evaluation model allowed us to compare the efficiency of currently available valve prostheses, and among these, the SJM Regent and the Sorin Bicarbon Slimline exhibited the best performances. PMID- 15354109 TI - Initial experience with an endoscopic radial artery harvesting technique. AB - OBJECTIVE: The purpose of the study was to investigate the feasibility of an endoscopic radial artery harvesting technique to improve esthetic results and possibly reduce the incidence of neurologic complications observed with the open method. METHODS: Between July 1, 2002, and October 1, 2003, a total of 54 patients underwent endoscopic radial artery harvesting at our institution. Standard endoscopic equipment and a Harmonic Scalpel (Ethicon Endo-Surgery, Inc, Cincinnati, Ohio) were used. Mean age of the patients was 63 +/- 8.1 years, and 16% were female. All patients underwent a preoperative Allen test and duplex ultrasonography to demonstrate adequate ulnar collateral flow. The nondominant arm was used for radial artery harvesting. Mean clinical follow-up was 13 +/- 4.6 months. RESULTS: The artery was harvested through a 3-cm incision at the wrist and was divided at the elbow either through a small counterincision (n = 16) or endoscopically with the use of clips, Endoloop, and endoscopic scissors (n = 38). Mean harvest time was 42.2 +/- 16.9 minutes but decreased from 85 minutes for the first cases to 25 minutes for the last 5 cases. Mean harvested length was 19.6 +/ 1.7 cm. Harvesting complications included 1 conversion, 2 postoperative hematomas, 1 injury, 8 endoscopically controlled bleedings, and 15 cases of at least some superficial radial nerve paresthesia at 6 weeks (clinically relevant in 4 cases). Selective postoperative angiography revealed 1 occluded graft and 1 stenotic graft requiring percutaneous transluminal coronary angioplasty of the native vessel. Eighty-seven percent of the patients were enthusiastic about this new procedure. CONCLUSIONS: Endoscopic radial artery harvesting is a feasible procedure that requires a definite learning curve. Although nerve paresthesias were not completely eliminated in our experience, the technique provided ample patient satisfaction. Further clinical follow-up is required to determine long term patency rates. PMID- 15354110 TI - Amniotic air insufflation during minimally invasive fetoscopic fetal cardiac interventions is safe for the fetal brain in sheep. AB - BACKGROUND: Amniotic air insufflation during experimental fetoscopic fetal cardiac interventions greatly improves the visualization of intra-amniotic contents. The purpose of this study was to assess any histologically discernible effects from this approach on the fetal brain after short-term studies and long term survival in sheep. METHODS: Thirty pregnant ewes between 80 and 110 days of gestation underwent amniotic air insufflation during various fetoscopic fetal cardiac interventions. After 18 short-term and 12 long-term studies, the brains of the operated fetuses and-if available-their unoperated siblings were examined for hemorrhage, embolism, infarctions, inflammatory changes, and abnormal cortical maturation. RESULTS: Amniotic air insufflation during minimally invasive fetoscopic fetal cardiac interventions did not result in any histologically discernible damage to the brain in short-term and long-term studies in any but 2 sibling sheep. In the 2 affected siblings, a small area of chronic cortical frontal lobe infarction was observed after long-term survival. CONCLUSIONS: Amniotic air insufflation during minimally invasive percutaneous fetoscopic fetal cardiac interventions is safe for the fetal brain and does not compromise maternal hemodynamics in sheep. These findings encourage further investigation of the role this technique might play during fetoscopic fetal cardiac interventions in humans. PMID- 15354111 TI - Biodegradable polymer with collagen microsponge serves as a new bioengineered cardiovascular prosthesis. AB - OBJECTIVE: Biodegradable materials with autologous cell seeding have attracted much interest as potential cardiovascular grafts. However, pretreatment of these materials requires a complicated and invasive procedure that carries the risk of infection. To avoid these problems, we sought to develop a biodegradable graft material containing collagen microsponge that would permit the regeneration of autologous vessel tissue. The ability of this material to accelerate in situ cellularization with autologous endothelial and smooth muscle cells was tested with and without precellularization. METHODS: Poly(lactic-co-glycolic acid) as a biodegradable scaffold was compounded with collagen microsponge to form a vascular patch material. These poly(lactic-co-glycolic acid)-collagen patches with (n = 10) or without (n = 10) autologous vessel cellularization were used to patch the canine pulmonary artery trunk. Histologic and biochemical assessments were performed 2 and 6 months after the implantation. RESULTS: There was no thrombus formation in either group, and the poly(lactic-co-glycolic acid) scaffold was almost completely absorbed in both groups. Histologic results showed the formation of an endothelial cell monolayer, a parallel alignment of smooth muscle cells, and reconstructed vessel wall with elastin and collagen fibers. The cellular and extracellular components in the patch had increased to levels similar to those in native tissue at 6 months. CONCLUSIONS: The poly(lactic-co glycolic acid)-collagen microsponge patch with and without precellularization showed good histologic findings and durability. This patch shows promise as a bioengineered material for promoting in situ cellularization and the regeneration of autologous tissue in cardiovascular surgery. PMID- 15354112 TI - Primary closure for postoperative mediastinitis in children. AB - OBJECTIVES: Mediastinitis affects approximately 1% of children undergoing median sternotomy. Conventional therapy involves debridement followed by open wound care with delayed closure, days to weeks of closed suction or antimicrobial irrigation, and vacuum-assisted closure or muscle flap closure. We hypothesized that primary closure without prolonged suction or irrigation is an effective, less traumatic treatment for mediastinitis in children. METHODS: From January 1986 to July 2002, 6705 procedures involving median sternotomy were performed at the C. S. Mott Children's Hospital, resulting in 57 cases of mediastinitis (0.85%). Cases were divided into 2 groups, with 42 cases treated with primary closure and 15 cases treated with delayed or muscle flap closure. The 42 cases of primary closure comprised the primary study group of this institutional review board-approved, retrospective analysis. Patient demographics, surgical variables, mediastinitis-related parameters, and outcomes were evaluated. RESULTS: One patient had recurrent mediastinitis for an overall infection eradication rate of 97% (40/41). Three patients (7%) required re-exploration for suspected ongoing infection. Of these re-explorations, 1 patient had evidence of continued mediastinitis. The remaining 2 patients with sepsis of unclear cause had no clinical or culture evidence of recurrent infection. One of these patients ultimately died of sepsis without active mediastinitis for a hospital survival of 97% (41/42). No significant differences could be detected between the treatment successes and failures in this small cohort of patients. CONCLUSIONS: Simple primary closure is an effective means to treat selected cases of postoperative mediastinitis in children. The results compare favorably with other more lengthy or debilitating treatments. PMID- 15354113 TI - Worse early outcomes in women after coronary artery bypass grafting: is it simply a matter of size? PMID- 15354114 TI - Something to expect, diagnose, and treat early. PMID- 15354115 TI - Assessing the value of neoadjuvant chemoradiotherapy and pathologic downstaging in the treatment of non-small cell lung cancer. PMID- 15354116 TI - Studying the lumen in composite Y internal thoracic artery-saphenous vein grafts. PMID- 15354119 TI - Education from the heartland. PMID- 15354117 TI - Brain metastases. PMID- 15354120 TI - EMS manager as an agent of change. PMID- 15354121 TI - Just like bugs: IVs can be easy-even when there's no BP. PMID- 15354123 TI - Atrial flutter with left bundle branch block: putting it all together. PMID- 15354125 TI - Winning combination: how to excel at an ALS competition. PMID- 15354126 TI - Coming to America: Center for Emergency Medicine attracts EMS students from around the world. PMID- 15354127 TI - By the numbers: prehospital cardiac arrest research. PMID- 15354128 TI - Health educators in the community: a new role for EMTs? PMID- 15354129 TI - Measuring & interpreting vital signs: an in-depth discussion of the objective data points that lead to a presumptive diagnosis & treatment plan. PMID- 15354130 TI - The oxylator: an innovative oxygen delivery device. PMID- 15354132 TI - eJEMS: Electronic & data management issues: EPAD: Emergency Provider Access Directory. PMID- 15354133 TI - Street pizza. PMID- 15354134 TI - The last dance. PMID- 15354135 TI - Denver paramedics mistreat patients & coworkers. PMID- 15354144 TI - Is coral calcium a safe and effective supplement? PMID- 15354145 TI - Bariatric surgery: life-threatening risk or life-saving procedure? PMID- 15354146 TI - Keeping up with a continuing trend in foodservice. PMID- 15354147 TI - Matching funds from the federal government for Medicaid disease management: dietitians as recognized providers. PMID- 15354148 TI - Total parenteral nutrition-induced hyperglycemia in hematopoietic stem cell transplantation. PMID- 15354149 TI - The incidence of hyperglycemia in hematopoietic stem cell transplant recipients receiving total parenteral nutrition: a pilot study. AB - OBJECTIVE: To determine whether total parenteral nutrition (TPN)-induced hyperglycemia is associated with adverse clinical outcomes. DESIGN: A retrospective cohort investigation comparing the medical records of hematopoietic stem cell transplant patients was conducted to determine clinical differences between those who received TPN and those who did not receive TPN during transplant. SUBJECTS/SETTING: Forty-eight adult patients (> or =18 years) undergoing initial autologous or allogeneic hematopoietic stem cell transplant at two urban university-affiliated hospitals were eligible for inclusion. MAIN OUTCOME MEASURES: Hyperglycemia (glucose > or =6.1 mmol/L or 110 mg/dL), presence of infection, infection duration, and in-hospital mortality. Statistical analyses performed chi 2, Student t, and Wilcoxon rank-sum tests were used to detect differences among the study participants. RESULTS: Patients had similar baseline demographic and clinical characteristics, with 63% receiving TPN during transplant. When standardized for time, TPN recipients at both institutions experienced significantly more hyperglycemia ( P <.05) after TPN initiation. TPN patients also experienced 69% of all infections and 100% of repeat positive cultures. Additionally, significantly greater differences for TPN recipients were found for length of stay and daily charges than those who did not receive TPN. No differences were found for in-hospital mortality. CONCLUSIONS: TPN is strongly associated with hyperglycemia, which may be linked to increased infections of longer duration in a profoundly immunocompromised group of patients who frequently receive TPN. The implications of these findings are limited by the small number of subjects; a larger investigation is warranted. PMID- 15354150 TI - Girls on a high-calcium diet gain weight at the same rate as girls on a normal diet: a pilot study. AB - OBJECTIVE: To determine whether pubertal girls assigned to calcium-rich diets or their usual calcium intakes differ significantly in weight gain. DESIGN: This is a pilot study conducted on data from an experimental research project. SUBJECTS/SETTING: The sample included 59 girls, 9 years of age, from Omaha metropolitan communities. INTERVENTION: Participants were randomly assigned to a calcium-rich diet supplying at least 1,500 mg of calcium per day or their usual diet. MAIN OUTCOME MEASURE: The outcome measure was change in weight during 2 years of study. STATISTICAL ANALYSES: Data were analyzed using descriptive statistics, Mann-Whitney U, t tests of paired and independent samples, and analysis of covariance. RESULTS: The girls in the calcium-rich diet group had a mean (+/-standard deviation) calcium intake of 1,656+/-191 mg/day, whereas the girls on their usual diets averaged 961+/-268 mg/day. Although the participants in the treatment group consumed nearly twice as much dietary calcium--primarily from dairy foods--they did not have greater increases in body weight, body mass index, or fat or lean mass than the control group. These findings held when the data were grouped by tertile of calcium intake. Compared with girls on their usual diets, the girls who consumed the calcium-rich diet also significantly increased their intake of essential nutrients. CONCLUSION: We conclude that calcium-rich diets do not cause excessive weight gain in pubertal girls but do contribute positively to overall nutrition. These findings provide support for programs to promote calcium-rich diets, which are critical for attaining peak bone mass. PMID- 15354151 TI - Possible benefits of a calcium-rich diet for girls. PMID- 15354152 TI - Dieting practices in high school students. AB - OBJECTIVE: The purpose of this research was to determine the patterns and frequency of dieting behaviors in a high school population in a metropolitan area. DESIGN: Dieting practices were assessed by using a survey instrument consisting of demographic questions and questions regarding dieting practices. SUBJECTS/SETTING: The survey was administered to 10th-grade male and female students at a multiethnic, urban, public high school in the Los Angeles area. One hundred fifty-three surveys were distributed in the classroom setting, with 146 usable surveys analyzed. STATISTICAL ANALYSES: Statistical analyses included simple frequencies, chi 2, Fisher exact test, t tests, and two-way analysis of variance. A significance level of.05 was used for all statistical analyses. RESULTS: The population with a body mass index greater than 25 was 26.6% and, therefore, at risk of being overweight. Of those who have tried dieting, 54.7% often diet to control their weight. Limiting portion size was practiced by about 34% of those who had tried dieting techniques. In addition, counting calories and counting grams of fat were reported by 31.4% and 41.9% of these students, respectively. Of interest, 64.8% of the students who had tried dieting reported that they tried to eat and/or purchase foods that were low in fat. Approximately, 44% of these students used meal skipping to control their weight. CONCLUSIONS: The high prevalence of dieting behaviors suggests that urban adolescents should be reached with appropriate interventions. Safe dieting strategies should be an integral part of any nutrition education and weight loss intervention. PMID- 15354153 TI - Modifying the Healthy Eating Index to assess diet quality in children and adolescents. AB - OBJECTIVE: The Healthy Eating Index (HEI) is a scoring system used by the US government to assess adherence to the Dietary Guidelines for Americans. We examined the ability of the HEI to monitor diet quality among youth. DESIGN: We modified and simplified the HEI for use by older children and adolescents. The new Youth Healthy Eating Index (YHEI) focuses on food quality and assesses both healthful and unhealthful foods and eating behaviors. Both HEI and YHEI scores were calculated from a food frequency questionnaire that was mailed to participants in the Growing Up Today Study in 1996. SUBJECTS/SETTING: Girls (n=8,807) and boys (n=7,645) 9 to 14 years of age who are children of participants in the Nurses Health Study II cohort and who reside across the United States. STATISTICAL ANALYSIS: Mean HEI and YHEI scores were calculated by sex and age, and associations with age, body mass index, activity, inactivity, energy intake, and several nutrients were assessed with Pearson correlations. Linear regression was used to examine the contributions of the individual HEI and YHEI components toward the total scores. RESULTS: The HEI score was highly correlated with total energy intake ( r =0.67), indicating a strong association with quantity of food consumption. In contrast, the YHEI was not strongly correlated with energy intake ( r =0.12) but was inversely associated with time spent in inactive pursuits ( r =-0.27). The HEI component for variety in food selection accounted for 60% of the variation in the total score and several HEI components were highly correlated with each other, particularly those for total and saturated fat ( r =0.78). CONCLUSIONS: To successfully monitor diet in a population of children and adolescents, the HEI may benefit from modifications that focus on food quality and include assessments of unhealthful foods. Further research is needed to determine the dietary elements that are most related to health in diverse populations of youth. PMID- 15354154 TI - Assessing diet quality in children and adolescents. PMID- 15354156 TI - Reemergence of pica following gastric bypass surgery for obesity: a new presentation of an old problem. AB - Abstract Pica, the compulsive ingestion of nonnutritive substances, has been a fascinating and poorly understood phenomenon for centuries. Pagophagia, or ice eating, is one of the most common forms of pica and is closely associated with the development of iron-deficiency anemia. Although this condition has been well described among pregnant women and malnourished children, particularly in developing countries, it has not been previously reported to occur following gastric bypass surgery for treatment of severe obesity. This article presents two cases of women who experienced a recurrence of pagophagia following gastric bypass surgery, along with an updated review of the literature. PMID- 15354155 TI - Assessment of interobserver reliability in nutrition studies that use direct observation of school meals. AB - This article (a) provides a general review of interobserver reliability (IOR) and (b) describes our method for assessing IOR for items and amounts consumed during school meals for a series of studies regarding the accuracy of fourth-grade children's dietary recalls validated with direct observation of school meals. A widely used validation method for dietary assessment is direct observation of meals. Although many studies utilize several people to conduct direct observations, few published studies indicate whether IOR was assessed. Assessment of IOR is necessary to determine that the information collected does not depend on who conducted the observation. Two strengths of our method for assessing IOR are that IOR was assessed regularly throughout the data collection period and that IOR was assessed for foods at the item and amount level instead of at the nutrient level. Adequate agreement among observers is essential to the reasoning behind using observation as a validation tool. Readers are encouraged to question the results of studies that fail to mention and/or to include the results for assessment of IOR when multiple people have conducted observations. PMID- 15354157 TI - America's obesity epidemic: measuring physical activity to promote an active lifestyle. AB - The incidence of overweight and obesity in the United States and worldwide has reached epidemic proportions. To effectively intervene, dietetics professionals and other health care practitioners need to address both sides of the energy balance equation when counseling clients and patients. Often, the focus on energy intake supersedes the promotion of a physically active lifestyle. Incorporating appropriate and sufficient physical activity into one's life is an essential component of achieving and maintaining a healthful body weight. This review summarizes background knowledge on the benefits of physical activity for health and provides an overview of available tools for measuring physical activity and energy expenditure. The physical and mental health benefits of an active lifestyle, current physical activity recommendations for the US public, the prevalence of inactivity in the United States, and components of energy expenditure are reviewed. Additionally, tools for estimating total energy expenditure, resting metabolic rate, and physical activity are evaluated and suitable approaches for applying these tools are provided. PMID- 15354158 TI - Dietary phylloquinone intake as a potential marker for a heart-healthy dietary pattern in the Framingham Offspring cohort. AB - Abstract Associations were evaluated among self-reported dietary intakes of phylloquinone (vitamin K-1), lifestyle characteristics, and intermediary markers of cardiovascular disease risk in a population-based cohort of men and women. Dietary phylloquinone intakes were assessed by food frequency questionnaire in 1,338 men and 1,603 women (mean age, 54 years) participating in the Framingham Heart Study. Cross-sectional associations with lifestyle characteristics and lipid profiles, including total cholesterol, low-density lipoprotein and high density lipoprotein cholesterol, and triglyceride concentrations, were estimated across increasing quintile categories of phylloquinone intakes. Participants in the highest quintile category of phylloquinone intake consumed more fruit, vegetables, fish, dietary fiber, and dietary supplements ( P <.001), and consumed less meat and less saturated fat ( P <.001). Higher phylloquinone intakes were also associated with lower triglyceride concentrations ( P <.001). In conclusion, a high phylloquinone intake may be a marker for an overall heart-healthy dietary pattern. PMID- 15354159 TI - The youth/adolescent questionnaire has low validity and modest reliability among low-income African-American and Hispanic seventh- and eighth-grade youth. AB - The Youth/Adolescent Questionnaire is a food frequency questionnaire developed for youth aged 9 to 19 years that attempts to assess diet during the past 12 months. It was developed and previously validated with white youth; however, a validation study with fourth- to seventh-grade African-American youth revealed lower validation coefficients. This study validated the Youth/Adolescent Questionnaire with 89 low-income seventh- and eighth-grade Hispanic and African American youth. The youth completed the Youth/Adolescent Questionnaire on two occasions 3 weeks apart, and completed up to six food records in between. Most of the reliability coefficients were adequate, except for percent of energy from fat and high-fat vegetables for Hispanic students. Validity coefficients were low, even after correcting for error. The ability of seventh- and eighth-grade low income African-American and Hispanic youth to report usual intake over a period of time appears to be limited and may not provide a valid measure of consumption. PMID- 15354160 TI - Commercial portion-controlled foods in research studies: how accurate are label weights? AB - OBJECTIVE: The purpose of this study was to evaluate the reliability of label weights as surrogates for actual weights in commercial portion-controlled foods used in a research setting. DESIGN: Actual weights of replicate samples of 82 portion-controlled food items and 17 discrete units of food from larger packaging were determined over time. Comparison was made to the package label weights for the portion-controlled food items and the per-serving weights for the discrete units. SETTING: The study was conducted at the US Department of Agriculture's Beltsville Human Nutrition Research Center's Human Study Facility, which houses a metabolic kitchen and human nutrition research facility. MAIN OUTCOME MEASURES: The primary outcome measures were the actual and label weights of 99 food items consumed by human volunteers during controlled feeding studies. Statistical analyses performed The difference between label and actual weights was tested by the paired t test for those data that complied with the assumptions of normality. The Wilcoxon signed rank test was used for the remainder of the data. Compliance with federal guidelines for packaged weights was also assessed. RESULTS: There was no statistical difference between actual and label weights for only 37 food items. The actual weights of 15 portion-controlled food items were 1% or more less than label weights, making them potentially out of compliance with federal guidelines. CONCLUSIONS: With advance planning and continuous monitoring, well controlled feeding studies could incorporate portion-controlled food items and discrete units, especially beverages and confectionery products. Dietetics professionals should encourage individuals with diabetes and others on strict dietary regimens to check actual weights of portion-controlled products carefully against package weights. PMID- 15354161 TI - Position of the American Dietetic Association and Dietitians of Canada: nutrition intervention in the care of persons with human immunodeficiency virus infection. AB - Infection with the human immunodeficiency virus (HIV) and the development of acquired immunodeficiency syndrome (AIDS) have had a significant impact on domestic and global health, social, political, and economic outcomes. Prevention and treatment efforts to control HIV infection are more demanding than in previous decades. Achieving food and nutrition security, and managing nutrition related complications of HIV infection and the multiple aspects of disease initiated by or surrounding HIV infection, referred to as HIV disease, remain challenges for patients and for those involved with HIV/AIDS prevention, care, and treatment efforts. Confounding clinical issues include medication interactions, coinfection with other infections and diseases, wasting, lipodystrophy, and others. Dietetics professionals, other health care professionals, and people infected with HIV will need to understand and address multiple complex aspects of HIV infection and treatment to improve survival, body functions, and overall quality of life. Individualized nutrition care plans will be an essential feature of the medical management of persons with HIV infection and AIDS. PMID- 15354175 TI - Chronic lithium treatment of B lymphoblasts from bipolar disorder patients reduces transient receptor potential channel 3 levels. AB - Chronic lithium treatment of B-lymphoblast cell lines (BLCLs) from bipolar-I disorder (BD-I) patients and healthy subjects ex vivo attenuates agonist- and thapsigargin-stimulated intracellular calcium (Ca(2+)) responses. As these findings suggest that chronic lithium treatment modifies receptor (ROCE) and/or store-operated Ca(2+) entry (SOCE) mechanisms, we determined whether chronic lithium treatment of BLCLs modified the expression of two members of the transient receptor potential channels (TRPC1 & 3), which participate in ROCE/SOCE. Chronic lithium treatment significantly reduced BLCL TRPC3 immunoreactivity (repeated-measures ANOVA, P=0.00005), with interaction effects of diagnosis (P=0.037) and sex (P=0.040). The lithium-induced decrease was greatest in BLCLs from female BD-I patients compared with those from healthy females (-27%) and with vehicle-treated BLCLs from female BD-I patients (-33%). However, lithium treatment did not affect TRPC1 and 3 mRNA levels, and TRPC1 immunoreactivity. Downregulation of TRPC3 may be an important mechanism by which lithium ameliorates pathophysiological Ca(2+) disturbances as observed in BD. PMID- 15354176 TI - Differential display polymerase chain reaction reveals increased expression of striatal rat glia-derived nexin following chronic clozapine treatment. AB - Clozapine is considered a prototype of the 'so-called' atypical antipsychotic drug class. It has affinity for a broad range of receptors and, in comparison to typical antipsychotic drugs, produces less extrapyramidal side effects. However, its mechanism of action remains unclear. Differential display polymerase chain reaction (ddPCR) was implemented in this study to contribute to the current understanding of this mechanism at the genetic level and to identify novel genes regulated by clozapine. This technique generated approximately 2400 gene sequences that were analyzed for differential gene expression following protracted clozapine treatment. One of these sequences, originally termed Clozapine Regulated Gene (CRG), was shown to be significantly upregulated following the treatment. Northern hybridization confirmation of this finding revealed that chronic clozapine administration caused a five-fold increase in CRG mRNA. Elongation of the 5'- and 3'-ends of CRG indicated that the fragment was in fact rat glia-derived nexin mRNA. Western blotting demonstrated that levels of the mRNA's associated protein also increased comparably (three-fold) following chronic treatment with the antipsychotic drug. This study presents a possible neuroprotective role of nexin in clozapine treatment, particularly in the prevention of neuronal proteolytic degradation, since nexin has been shown to be a protease inhibitor. PMID- 15354177 TI - Combined actions of zinc and fluoxetine on nicotinic acetylcholine receptors. AB - Zinc and nicotinic acetylcholine receptors (nAChRs) seem to be associated with major depression, and some antidepressants, including fluoxetine (Prozac), antagonize nAChRs. Therefore, a study was made of the modulation of neuronal alpha4beta4 and muscle alpha1beta1gammadelta nAChRs, expressing in oocytes, by the combined action of zinc and fluoxetine. At a holding potential of -60 mV, 200 microM zinc increased by 361% the currents elicited by acetylcholine (ACh currents) for alpha4beta4 and by 182% for alpha1beta1gammadelta nAChRs. In contrast, 5 microM fluoxetine reduced the ACh currents to 31% for alpha4beta4 and to 45% for alpha1beta1gammadelta nAChRs. Additionally, fluoxetine reduced more the ACh currents in the presence of zinc: to 17% for alpha4beta4 and to 19% for alpha1beta1gammadelta nAChRs, and after washing out the fluoxetine the ACh current did not recover its zinc-potentiated value. Moreover, when ACh-activated nAChRs were exposed first to fluoxetine and then zinc was added, the potentiating effect of zinc was very small for muscle nAChRs and was nil for neuronal receptors. Thus, the inhibiting effect of fluoxetine prevails over the potentiating action of zinc. Finally, the effects of both zinc and fluoxetine were voltage independent, indicating that these substances interact outside the ion channel. As fluoxetine nullifies the effects of zinc, it appears that both substances interact in the same site. These results should help understand better the roles played by zinc, antidepressants, nAChRs and their combination in brain functions and in the treatment of depression. PMID- 15354178 TI - Loss of expression of bone morphogenetic protein receptor type II in human prostate cancer cells. AB - Bone morphogenetic proteins (BMPs) are members of the transforming growth factor beta superfamily and signal through a number of membrane receptors. We have previously demonstrated that the loss of expression of BMP receptors (BMPRs) type IA, -IB, and -II (BMP-RIA, -RIB, and -RII) correlates with Gleason score in prostate cancer patients. To evaluate the prognostic value of this observation, we used immunohistochemistry to investigate the expression of BMPRs in association with disease progression in 60 patients. The results demonstrated a significant association between the loss of expression of the three BMPRs and Gleason score and clinical stage. However, only the loss of expression of BMP-RII showed a statistically significant association with 5-year survival rate (P<0.05) and biochemical recurrence-free rate following radical prostatectomy (P<0.005). To elucidate the effect of an abnormal BMP signaling in prostate cancer cells, we transfected dominant-negative BMP-RII (BMP-RIIDN) into the human prostate cancer cell line, PC3M. When a stable clone overexpressing BMP-RIIDN was inoculated subcutaneously into nude mice, the tumor growth rate was approximately 10 times that of control and parental cell line. These observations, taken together, indicate that the loss of BMP-RII expression as measured by immunohistochemistry may be a prognostic marker in prostate cancer patients, and that the loss of BMP RII function may result in increased tumorigenicity in human prostate cancer cells. PMID- 15354179 TI - Is there a definition of remission in late-life depression that predicts later relapse? AB - Remission of depressive symptoms is the goal of all antidepressant therapy. Rating scales define remission in clinical trials, but it is unclear how well these definitions predict risk of later relapse. We measured the sensitivity and specificity of a range of Montgomery-Asberg Depression Rating Scale (MADRS) cutoff scores at 3- and 6-months, wherein scores above a given cutoff would predict relapse over an 18-month period. We examined 153 elderly depressed subjects exhibiting a MADRS < or = 15 after 3 or 6 months of antidepressant therapy. Subjects who subsequently exhibited a MADRS > 15 during the 18-month study period were defined as relapsed. Receiver operating characteristic (ROC) curves were developed and area under the curve (AUC) values calculated for the sensitivity and specificity of 3- and 6-month MADRS scores to predict future relapse. The 3-month ROC had an AUC value of 0.63; the 6-month ROC had an AUC value of 0.66. There was no MADRS cutoff found that could predict likelihood of relapse with good sensitivity and specificity. A post hoc analysis where relapse rate was adjusted by controlling for medical comorbidity, disability, and social support showed no change in the ROCs or AUC values. The higher the MADRS score at 3 and 6 months, the greater the likelihood of relapse. With no clean MADRS cutoff score, the goal of antidepressant therapy should be the lowest possible degree of depressive symptomatology to minimize risk of later relapse. Definitions of remission that are better associated with longer-term outcomes are needed. PMID- 15354180 TI - Effects of serotonin transporter promoter polymorphisms on serotonin function. AB - The serotonin transporter promoter polymorphism (5-HTTLPR) has been associated with vulnerability to stress-induced depressive symptoms and with the speed and rate of response to antidepressant treatment. The goal of the present study was to evaluate the association between the 5-HTTLPR and the functional response of the serotonin system as measured by the neuroendocrine and cerebral metabolic response to intravenous administration of the selective serotonin reuptake inhibitor citalopram in normal control subjects. Genotyping was performed for 5 HTTLPR insertion/deletion polymorphism long (l) and short (s) variant alleles. The ll genotype was compared with the combined sl+ss and with the ss genotype alone. Citalopram plasma concentrations did not differ significantly between groups. The s allele was associated with a less of an increase in prolactin and cortisol than the ll genotype. The s allele was associated with greater decreases in left frontal, precentral and middle temporal gyri compared to the ll genotype. The ll genotype was associated with greater decreases in right frontal, insula and superior temporal gyrus compared to the ss genotype. These findings suggest that 5-HTTLPR is associated with an altered functional response of the serotonin system, which may represent a neurobiologic substrate for the differential response to antidepressant treatment in late life and the emergence of neuropsychiatric symptoms in neurodegenerative disorders. PMID- 15354181 TI - Brain metabolites and cognitive function among older depressed and healthy individuals using 2D MR spectroscopy. AB - Brain metabolites of choline (Ch) and myo-Inisotol (mI) have been reported as elevated among geriatric depressed patients. Two-dimensional (2D) magnetic resonance spectroscopy (MRS) provides estimates of Ch, mI, and creatine (Cr) similar to one-dimensional MRS, and it also estimates the resonances of the Ch containing compounds of phosphoethanolamine (Pe) and phosphocholine (PCh). In this cross-sectional geriatric study, 14 depressed patients and 14 healthy volunteers who were comparable in age, gender, education, comorbid medical burden, and Mini-Mental State Examination (MMSE) scores completed 2D MRS and a neurocognitive battery. A voxel in the left dorsolateral cortex, which was comprised of approximately 60% white matter, was used to estimate the CR ratios of Ch, PCh, Pe, and mI. Composite scores for cognitive function were developed for verbal learning, recall, recognition, executive function, hypothesis generation, and processing speed. Among nondepressed subjects, cognition was positively correlated with Ch/Cr and mI/Cr and negatively correlated with PCh/Cr in four domains of verbal learning, recognition, recall, and hypothesis generation. In contrast, depressed patients did not have consistent relationships between Ch/Cr, mI/Cr, and PCh/Cr and cognition. There was a significant difference in the overall pattern of associations between the four metabolites and verbal learning and processing speed in depressed patients compared to healthy controls. The attenuated relationship between metabolites and specific cognitive domains in patients with late-life MDD suggests that the level of cognitive performance observed during depressive episodes may be associated with changes in biochemistry within the frontostriatal neuronal circuitry. PMID- 15354182 TI - Neuropathologic correlates of late-onset major depression. AB - Late life major depression (LLMD) is frequently associated with cognitive impairment, and increases the risk for subsequent dementia. Cerebrovascular disease, Alzheimer's disease (AD), and dementia with Lewy bodies (DLB) have all been hypothesized to contribute to this increased risk, though prospective studies have yet to examine these hypotheses with autopsy confirmation of the clinical diagnoses. The aim of this study is to examine the rates of cerebrovascular, AD, and DLB pathology among the first 10 participants in an LLMD brain tissue donation program. Subjects' psychiatric diagnoses and cognitive status were prospectively determined during their participation in clinical research protocols of the Intervention Research Center for Late Life Mood Disorders. After death, final clinical diagnoses were made using all clinical information, while blind to neuropathologic diagnoses. Neuropathologic assessments were conducted blind to final clinical diagnoses. Rates of neuropathology were compared with those in a cohort of subjects with dementia, without a history of LLMD, participating in an Alzheimer Disease Research Center. Seven (70%) subjects had evidence of onset of a dementia prior to death. LLMD with dementia was significantly associated with a neuropathologic diagnosis of AD. Cerebrovascular disease and DLB pathology were also frequent in the LLMD subjects with dementia, and were found in an LLMD subject without dementia. Rates of AD, DLB, and cerebrovascular disease were similar to those in the comparison subjects. These preliminary findings suggest that AD is the predominant neuropathologic condition in LLMD subjects with dementia. Further assessment of the role of comorbid cerebrovascular disease and comorbid DLB is needed. PMID- 15354183 TI - Cerebrospinal anandamide levels are elevated in acute schizophrenia and are inversely correlated with psychotic symptoms. AB - The endocannabinoids are a family of bioactive lipids that activate CB1 cannabinoid receptors in the brain and exert intense emotional and cognitive effects. Here, we have examined the role of endocannabinoid signaling in psychotic states by measuring levels of the endocannabinoid anandamide in cerebrospinal fluid (CSF) of acute paranoid-type schizophrenic patients. We found that CSF anandamide levels are eight-fold higher in antipsychotic-naive first episode paranoid schizophrenics (n = 47) than healthy controls (n = 84), dementia patients (n = 13) or affective disorder patients (n = 22). Such an alteration is absent in schizophrenics treated with 'typical' antipsychotics (n = 37), which antagonize dopamine D2-like receptors, but not in those treated with 'atypical' antipsychotics (n = 34), which preferentially antagonize 5HT(2A) receptors. Furthermore, we found that, in nonmedicated acute schizophrenics, CSF anandamide is negatively correlated with psychotic symptoms (rS = -0.452, P = 0.001). The results suggest that anandamide elevation in acute paranoid schizophrenia may reflect a compensatory adaptation to the disease state. PMID- 15354184 TI - NAAG reduces NMDA receptor current in CA1 hippocampal pyramidal neurons of acute slices and dissociated neurons. AB - N-acetylaspartylglutamate (NAAG) is an abundant neuropeptide in the nervous system, yet its functions are not well understood. Pyramidal neurons of the CA1 sector of acutely prepared hippocampal slices were recorded using the whole-cell patch-clamp technique. At low concentrations (20 microM), NAAG reduced isolated N methyl-D-aspartate receptor (NMDAR)-mediated synaptic currents or NMDA-induced currents. The NAAG-induced change in the NMDA concentration/response curve suggested that the antagonism was not competitive. However, the NAAG-induced change in the concentration/response curve for the NMDAR co-agonist, glycine, indicated that glycine can overcome the NAAG antagonism. The antagonism of the NMDAR induced by NAAG was still observed in the presence of LY-341495, a potent and selective mGluR3 antagonist. Moreover, in dissociated pyramidal neurons of the CA1 region, NAAG also reduced the NMDA current and this effect was reversed by glycine. These results suggest that NAAG reduces the NMDA currents in hippocampal CA1 pyramidal neurons. PMID- 15354185 TI - Constitutive hyperdopaminergia is functionally associated with reduced behavioral lateralization. AB - According to the dopamine (DA) hypothesis of schizophrenia and the strong evidence for decreased cerebral lateralization in schizophrenic patients, we postulated that hyperactivity of the dopaminergic system could be associated with a reduced behavioral lateralization in mice. Mice lacking the dopamine transporter (DAT) gene were used as a genetic model of persistent hyperdopaminergia. The DAT null mutation was transferred on C57BL/6JOrl (B6) and DBA/2JOrl (D2) inbred backgrounds for more than 10 generations of backcrossing to derive three DAT strains, B6, D2, and B6xD2(F1). Adult mutant mice of the three DAT strains and their littermates were tested for paw preference using Collins' protocol. Our results demonstrated that, whatever the genetic background, persistent hyperdopaminergia directly impairs the degree of lateralization without affecting the direction. Our results support the degree of lateralization as a good candidate phenotype to further improve genetic analysis of cerebral lateralization in normal and pathological conditions. PMID- 15354186 TI - Proliferation profile of classical Hodgkin's lymphomas. Increased expression of the protein cyclin D2 in Hodgkin's and Reed-Sternberg cells. AB - There is accumulating evidence that Hodgkin's and Reed-Sternberg cells of classical Hodgkin's lymphomas (cHL) display multiple and concurrent alterations in different pathways and checkpoints of the cell cycle. However, the expression of cyclin D2 and its relation to other major cell cycle proteins has not been analyzed in cHL. The aim of the present study was to assess expression of cyclin D2, Ki67, cyclin A, cyclin B1, cyclin D1, cyclin D3, cyclin E, p53, Rb, p16 and p27 proteins in order to gain further insight into the proliferation profile of cHL. Overexpression of cyclin D2 in Hodgkin's and Reed-Sternberg cells was detected in 64/89 (72%) cases of cHL. This finding, in view of recent in vitro data showing that constitutive activation of nuclear factor (NF)-kB could upregulate cyclin D2 expression in part via signal transducer and activator of transcription (STAT)-5a, suggests that induction of cyclin D2 expression may support the proliferation of Hodgkin's and Reed-Sternberg cells. In addition, the present study showed that (1) increased p27 expression status was significantly correlated with higher levels of cyclin A expression (P=0.048) and (2) increased p53 expression status was significantly correlated with higher levels of cyclin A (P<0.001) and cyclin B1 (P=0.040) expression. The association between increased p27 and p53 expression status and higher expression levels of G2/M cyclins suggests that the impairment of the growth inhibitory activity of the p27 and p53 tumor suppressor pathways may promote the proliferation of Hodgkin's and Reed Sternberg cells. PMID- 15354188 TI - Molecular and morphological differentiation between the crop and weedy types in velvetleaf (Abutilon theophrasti Medik.) using a chloroplast DNA marker: seed source of the present invasive velvetleaf in Japan. AB - A comparison of chloroplast DNA (cpDNA) sequences was carried out between the crop and weed types of Abutilon theophrasti to clarify the seed source of the present weedy velvetleaf in Japan. A sequencing analysis of approx. 6% of the chloroplast genome (ca 10 kbp) detected three nucleotide substitutions, one six base-pair insertion/deletion (indel) and one 30-base pair inversion, which distinguish two haplotypes of cpDNA. A PCR-based survey of the indel and the inversion revealed that the 93 accessions of velvetleaf collected from the world could be divided into two groups. A morphological marker (capsule color) could be used to discriminate the crop type and the weed type, and hence, along with cpDNA haplotype, to distinguish three genotypes (Type I, II, and III). All Japanese cultivars and crop accessions from other countries were Type I. Weed types were divided into Type II and III. All of the samples from the USA, and the samples taken from grain imports to Japan were Type III. Since most of the weedy types distributed in Japan were of Type III, it is argued that they were introduced as seeds in the imported grain. We also found that the Type II plants sporadically occurred in Japan. It is suggested that they originated as hybrids, with indigenous cultivars as the maternal ancestor. Such hybrids must have survived since the cessation of velvetleaf cultivation about a century ago. PMID- 15354187 TI - Defective mismatch repair in the pathogenesis of low-grade appendiceal mucinous neoplasms and adenocarcinomas. AB - Defective DNA mismatch repair has been proposed as a second pathway for colonic carcinogenesis, particularly in tumors arising in the right colon. We investigated whether tumors arising in the appendix are associated with defective DNA mismatch repair using immunohistochemistry for mismatch repair enzymes hMLH 1, hMSH-2, hMSH-6, and hPMS-2. These immunoassays have been shown to be highly sensitive and specific for defective DNA mismatch repair in sporadic and familial adenocarcinomas. Sporadic adenocarcinomas with defective DNA mismatch repair essentially always show loss of hMLH-1, while loss of hMSH-2, hMSH-6, or hPMS-2 is almost always due to germline mutation. In all, 35 cases of appendiceal epithelial neoplasms were evaluated, comprising 18 low-grade appendiceal mucinous neoplasms confined to the appendix; eight low-grade appendiceal mucinous neoplasms with extra-appendiceal spread (five peritoneum and ovaries, two peritoneum, one ovaries only); and nine invasive adenocarcinomas (three with metastatic disease). All immunohistochemical slides were reviewed by two pathologists. One (11%) invasive adenocarcinoma showed absent expression of hMSH 2 and hMSH-6, but preserved hMLH-1 and hPMS-2 expression. This case was a 26-year old female with a history of synovial sarcoma who presented with acute appendicitis and appendiceal perforation (median age for other invasive carcinomas, 62 years; range 38-76 years). The appendiceal tumor was a moderately differentiated, colonic-type adenocarcinoma without significant extracellular mucin or tumor-infiltrating lymphocytes. The remaining invasive carcinomas and low-grade appendiceal mucinous neoplasms demonstrated preserved expression of all mismatch repair enzymes, including the seven cases in which extra-appendiceal tumor was also evaluated. We conclude that defective DNA mismatch repair does not play a role in the pathogenesis of low-grade appendiceal mucinous neoplasms. Defective DNA mismatch was found in 11% of invasive carcinomas, likely due to a germline mutation. These findings suggest that sporadic appendiceal neoplasia rarely arises through the defective DNA mismatch repair (mutator) pathway. PMID- 15354189 TI - Quantitative trait loci and interaction effects responsible for variation in female postmating mortality in Drosophila simulans and D. sechellia introgression lines. AB - Mating appears to inflict a cost to Drosophila females, resulting in a reduction of their lifespan shortly after mating. Males from different chromosome extracted lines differ significantly in their detrimental effects on postmating female survival, and seminal fluid proteins produced in the male accessory glands are at least partially responsible for the effect. This suggests that there is a genetic basis underlying the male inflicted effect on female's postmating mortality. However, the genes responsible for this effect remain elusive. Using males from introgression lines between D. simulans and D. sechellia genomes and a quantitative trait locus (QTL) mapping approach, we identified chromosomal regions that affect postmating mortality of females. We found a second chromosome QTL with an effect on average female lifespan after mating and a third chromosome QTL with an effect on postmating female mortality rate. Under the general observation of a faster divergence of sex-related genes among closely related species, it is predicted that genes for reproductive traits other than hybrid sterility will show evidence of epistatic effects when brought into a heterospecific background. We detected a significant epistatic genetic effect on postmating female mortality rate that supports this prediction. PMID- 15354190 TI - Transcriptome evolution - much ado about nothing? PMID- 15354191 TI - Estimating the correlation of pairwise relatedness along chromosomes. AB - The 'spatial' pattern of the correlation of pairwise relatedness among loci within a chromosome is an important aspect for an insight into genomic evolution in natural populations. In this article, a statistical genetic method is presented for estimating the correlation of pairwise relatedness among linked loci. The probabilities of identity-in-state (IIS) are related to the probabilities of identity-by-descent (IBS) for the two- and three-loci cases. By decomposing the joint probabilities of two- or three-loci IBD, the probability of pairwise relatedness at a single locus and its correlation among linked loci can be simultaneously estimated. To provide effective statistical methods for estimation, weighted least square (LS) and maximum likelihood (ML) methods are evaluated through extensive Monte Carlo simulations. Results show that the ML method gives a better performance than the weighted LS method with haploid genotypic data. However, there are no significant differences between the two methods when two- or three-loci diploid genotypic data are employed. Compared with the optimal size for haploid genotypic data, a smaller optimal sample size is predicted with diploid genotypic data. PMID- 15354192 TI - Genetic architecture of resistance to aphids and mites in a willow hybrid system. AB - Hybrid plants often differ in resistance to arthropods compared to the parental species from which they are derived. To better understand the relative contribution of genetic effects in influencing plant resistance to arthropods, we examined the genetic architecture of resistance in a willow hybrid system, Salix eriocephala, S. sericea, and their interspecific hybrids. Resistance to two arthropods, a willow leaf aphid (Chaitophorus sp.: Aphididae) and an eriophyoid mite (Aculops tetanothrix: Eriophyidae), were compared because resistance to different herbivores may be controlled by different traits and influenced by different genetic effects. We found additive and nonadditive genetic effects to be important in explaining the difference between willow species in resistance to aphids and mites. F2 hybrids exhibited low resistance to aphids, suggesting breakdown of favourable epistatic interactions that confer resistance. F2 hybrids, however, exhibited high resistance to mites, suggesting either the breakdown of interactions that affect traits used by mites in host location or the creation of favourable epistatic interactions. This study demonstrates the potential role of herbivores in affecting plant genetic structure, such that selection by herbivores can potentially lead to the creation of gene interactions that influence host resistance traits or host recognition traits used by the herbivore. PMID- 15354193 TI - Effective population size in Hymenoptera with complementary sex determination. AB - Complementary sex determination in the haplodiploid Hymenoptera leads to the production of inviable or effectively sterile diploid males when diploid progeny are homozygous at the sex-determining locus. The production of diploid males reduces the number of females in a population and biases the effective breeding sex ratio in favor of haploid males. This in turn will reduce the effective population size (Ne) of hymenopteran populations with complementary sex determination relative to the expected reductions due to haplodiploidy alone. The effects of diploid male production on Ne in hymenopterans with complementary sex determination when diploid males are either inviable or effectively sterile are assessed theoretically. In both models, low allelic diversity at the sex locus reduces the Ne of hymenopteran populations, and this effect is largest when diploid males are effectively sterile. PMID- 15354194 TI - Genetic relationships between diploid and allotetraploid cherry species (Prunus avium, Prunus x gondouinii and Prunus cerasus). AB - Prunus avium L. (diploid, AA, 2n=2x=16), Prunus cerasus L. (allotetraploid, AAFF, 2n=4x=32) species, and their hybrid Prunus x gondouinii Rehd., constitute the most widely cultivated cherry tree species. P. cerasus is supposed to be an hybrid species produced by the union of unreduced P. avium gametes and normal P. fruticosa gametes. A continuum of morphological traits between these three species makes their assignation difficult. The aim of this paper is to study the genetic relationships between tetraploid and diploid cherry species. In all, 114 genotypes belonging to these species were analyzed using 75 AFLP markers. The coordinates of these genotypes on the first axis of a correspondence analysis allowed us to clearly distinguish each species, to identify misclassifications and to assign unknown genotypes to one species. We showed that there are specific alleles in P. cerasus, which are not present in the A genome of P. avium and which probably come from the F genome of P. cerasus. The frequencies of each marker in the A and the F genomes were estimated in order to identify A and F specific markers. We discuss the utility of these specific markers for finding the origin of the A and F genomes in the allopolyploid species. PMID- 15354195 TI - Genetic diversity of the shea tree (Vitellaria paradoxa C.F. Gaertn), detected by RAPD and chloroplast microsatellite markers. AB - RAPDs and chloroplast microsatellites were used to quantify the genetic variation of Vitellaria paradoxa (an economically important tree species in sub-Saharan Africa, north of the equator) and to analyse the geographic distribution of diversity in relation to the refuge theory. A total of 13 locations were sampled in eight countries, covering most of the natural range from Senegal to Uganda. A total of 67 polymorphic and 15 monomorphic RAPD loci were detected in 179 individuals. No relationship was identified between diversity and longitude or latitude. An unrooted neighbour-joining tree suggested a western group and an eastern group, representing 7% (P = 0.000) of the total variation. A Mantel test suggested that genetic distances between populations were correlated to geographic distances (R = 0.88, P = 0.001). The three-chloroplast microsatellite primers, assayed in 116 individuals, revealed 10 different alleles and seven chlorotypes. Most of the populations comprised a single haplotype. It is proposed from these results that the difference between western and eastern populations results from putative refugia separated by the current 'Dahomey Gap' (a semiarid zone that meets the coast around the Ghana-Togo-Benin-Nigeria borders), an area that may have been exceptionally dry during glacial periods. In addition, it is suggested that the haplotype distribution and frequency in the western populations could be due to the more recent impact of humans, particularly shea tree selection and dispersal during traditional agroforestry. PMID- 15354196 TI - Population structure and host-plant specialization in two Scaptodrosophila flower breeding species. AB - In contrast to phytophagous insect species, little attention has been paid to the possibility of host races in the Drosophilidae, although flower-breeding species, where courtship and mating take place on the flowers, are likely candidates. Two species of Scaptodrosophila, S. hibisci and S. aclinata, are restricted to flowers of Hibiscus species (section Furcaria), and the Furcaria specialization likely predated the separation of S. hibisci and S. aclinata. In all, 20 microsatellite loci were analysed in nine populations of S. hibisci and five of S. aclinata. For two pairs of S. hibisci populations in close proximity, but breeding on different Hibiscus species, differentiation between the populations of each of these pairs was similar to that between the populations that were from the same Hibiscus species, but geographically distant, suggesting the early stages of host-race formation. Genetic variability was significantly less in S. aclinata than in S. hibisci, suggesting greater drift effects in the former. However, of 253 alleles detected, 82 were present in both species, 160 in S. hibisci only and 11 in S. aclinata only, indicating that S. aclinata was derived from S. hibisci, following a strong bottleneck at the time of separation- possibly 40,000 years BP. Analyses and interpretation of Hardy-Weinberg equilibrium and F statistics needed to account for null alleles known to be present at eight loci in S. hibisci, and possibly present at other loci. The results emphasize the need for caution in studies where the presence of null alleles is inferred only from population data. PMID- 15354197 TI - Self-perception of physical competences in preadolescent overweight Chinese children. AB - OBJECTIVE: To compare self-perceptions of physical competences in overweight and in normal weight preadolescent Chinese children. DESIGN: Cross-sectional study. SETTING: Three primary schools and a university hospital in Hong Kong. SUBJECTS: A total of 634 children, comprising 558 (462 normal weight, 96 overweight) aged 8 12 y randomly sampled from three primary schools, and 76 similar age overweight children recruited from the community for a diet and exercise intervention programme. MEASUREMENTS: Height, weight and percentage body fat were measured. Self-perceptions of physical competences were determined by Physical Self Descriptive Questionnaire (PSDQ). Corresponding actual physical competences were measured by physical fitness tests. RESULTS: Overweight children perceived themselves to have significantly more body fat than normal weight children, with poorer appearance, sports competence, endurance, coordination, flexibility, overall physical self-concept and self-esteem, but to be no less healthy, no less physically active and no less strong. Overweight children performed less well than normal weight children in measures of endurance, coordination and flexibility but better in strength. Poor self-perception of physical competences appeared only partly related to deficiencies in actual physical competences. CONCLUSION: Overweight children have poorer self-perception of their physical competences but do not perceive themselves to be less strong, healthy or physically active than normal weight children. Exercise programmes for overweight children could be more effective if designed with the knowledge of these self perceptions. PMID- 15354198 TI - Infant feeding patterns in the Finnish type I diabetes prediction and prevention nutrition study cohort. AB - OBJECTIVE: To investigate infant feeding patterns during the first 2 y and their relation to sociodemographic factors. DESIGN: A population-based cohort study. SETTING: Oulu and Tampere University Hospital district areas 1996-1999, Finland. SUBJECTS AND METHODS: All newborn infants (n=675) with increased genetic risk for type I diabetes were invited to the study in 1996-1997. Of these, 429 (64%) completed the dietary follow-up form by the time they reached 2 y of age. RESULTS: The median duration of exclusive breastfeeding (BF) was 1.8 months (range 0-6 months) and that of total BF 7.0 months (0.3-25 months). Among the infants 20% were exclusively breastfed at least 4 months (recommendation 4-6 months). Infants were introduced to infant formula at the median age of 1.8 months (range 0-25 months) and other supplementary foods at the median age of 3.5 months (1-6 months). Infant's ponderal index at birth was inversely associated with the duration of total BF. The age of introduction of supplementary foods correlated positively with the duration of total BF. Longer parental education and increased maternal age were associated with a longer duration of BF and older age at introduction of supplementary foods. Infant formula and other supplementary foods were added earlier to the diet of the boys than that of the girls. CONCLUSION: Duration of breastfeeding in Finland is shorter than recommended. Compliance with the current recommendations on the timing of introduction of first supplementary food and dairy products is relatively poor. The diet during infancy seems to be conspicuously influenced by the duration of parental education, maternal age and the sex of the infant. PMID- 15354199 TI - Short-term effects of a very-low-protein diet supplemented with ketoacids in nondialyzed chronic kidney disease patients. AB - OBJECTIVE: To evaluate the effects on the nutritional and metabolic parameters of a very-low-protein diet supplemented with ketoacids (VLPD+KA) in comparison with a conventional low-protein diet (LPD) in chronic kidney disease (CKD) patients. DESIGN: Prospective, randomized, controlled clinical study. SETTING: Outpatient Clinic of the Nephrology Division of Federal University of Sao Paulo, Brazil. SUBJECTS: The study involved 24 patients with advanced CKD (creatinine clearance <25 ml/min) that were randomly assigned to either a VLPD+KA (VLPD+KA group, 12 patients) or to a conventional LPD with 0.6 g/kg/day (LPD group, 12 patients). The patients were followed for 4 months. RESULTS: Nutritional status was adequately maintained with both diets for the studied period. Protein intake and serum urea nitrogen decreased significantly only in the VLPD+KA group (from 0.68+/-0.17 to 0.43+/-0.12 g/kg/day, P<0.05; from 61.4+/-12.8 to 43.6+/-14.9 mg/dl, P<0.001; respectively). Ionized calcium did not change in the VLPD+KA group but tended to decrease in the LPD group. Serum phosphorus tended to decrease in the VLPD+KA group probably as a result of a significant reduction in dietary phosphorus (529+/-109 to 373+/-125 mg/day, P<0.05) associated to the phosphorus-binding effect of the ketoacids. No change in these parameters was found in the LPD group. Serum parathormone increased significantly only in the LPD group (from 241+/-138 to 494+/-390 pg/ml, P<0.01). The change in PTH concentration was negatively correlated with changes in ionized calcium concentration (r=-0.75, P=0.02) and positively correlated with changes in serum phosphorus (r=0.71, P=0.03) only in the LPD group. CONCLUSION: This study indicates that a VLPD+KA can maintain the nutritional status of the patients similarly to a conventional LPD. Besides, an improvement in calcium and phosphorus metabolism and a reduction in serum urea nitrogen were attained only with the VLPD+KA. Thus, VLPD+KA can constitute another efficient therapeutic alternative in the treatment of CKD patients. PMID- 15354200 TI - Amplification of the lytic potential of effector/memory CD8+ cells by vector based enhancement of ICAM-1 (CD54) in target cells: implications for intratumoral vaccine therapy. AB - We demonstrated that enhanced expression of the costimulatory molecules CD80, CD54 and CD48 (designated rF-TRICOM) on target cells, as delivered via a recombinant fowlpox vector, results in an increased state of stimulation of CD8+ T cells, and consequent increased lysis of target cells. CTL studies in conjunction with antibody-blocking studies demonstrated that the enhanced effector activity of these CD8+ T cells is mediated mainly through CD54. Intracellular staining of CD8+ cells that interact with target cells infected with rF-TRICOM showed that they contain higher amounts of perforin and have a higher level of perforin message. Enhanced expression of costimulatory molecules (specifically CD54) on target cells using rF-TRICOM vectors also leads to the formation of stable conjugates/synapses between targets and T cells. The interaction of T cells with target cells that overexpress costimulatory molecules upon infection with rF-TRICOM leads to enhanced signaling through Lck, ZAP70, and STAT-1 in CD8+ T cells and heightened lytic activity of CD8+ cells through the formation of a greater number of immunological synapses. This, in turn, leads to enhanced signaling in T cells. Finally, studies were conducted in mice in which CEA is a self-antigen in an attempt to understand the potential clinical relevancy of intratumoral vaccine therapy. Mice were transplanted subcutaneously with CEA expressing tumors. Intratumoral (i.t.) vaccination was administered 8 days post tumor transplant. Mice vaccinated i.t. with rF-TRICOM demonstrated significantly reduced tumor growth and 40% of the mice had complete tumor regression. The antitumor effects were further improved by the addition of tumor antigen (CEA) in the vaccination by utilizing rF-CEA/TRICOM, with 80% of the mice experiencing complete tumor regression. These studies thus support the concept of intratumoral vaccination employing vectors expressing costimulatory molecules. PMID- 15354201 TI - Death receptor 4 (DR4) efficiently kills breast cancer cells irrespective of their sensitivity to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). AB - Breast cancer cells are generally resistant to induction of apoptosis by treatment with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). In this study, we demonstrate that both TRAIL-sensitive and TRAIL-resistant breast cancer cell lines can be efficiently killed by overexpression of the TRAIL receptor, death receptor 4 (DR4). The extent of cell death depended on the strength of the promoter driving DR4 expression. When driven by the strong CMV promoter, expression of DR4 killed over 90% of cells in five out of six cell lines tested in the absence of exogenous TRAIL. When driven by the relatively weak tumor-specific hTERT promoter, DR4 was less effective alone, but sensitized cells to killing by TRAIL. The extent of TRAIL sensitization depended on the magnitude of hTERT promoter activity. MCF-7 cells were relatively resistant to the action of DR4. We compared expression of the genes involved in transduction and execution of the death receptor-initiated apoptotic stimuli between MCF-7 and DR4-sensitive cell lines. We confirmed that in the panel of cell lines, MCF-7 was the only line deficient in expression of caspase 3. Bcl-2 and FLIP proteins, implicated in suppression of TRAIL-induced apoptosis, were expressed at a higher level. PMID- 15354202 TI - Apoptosis and disease: a matter of cell fate. PMID- 15354203 TI - Antileukemic and long-term effects of two regimens with or without TBI in allogeneic bone marrow transplantation for childhood acute lymphoblastic leukemia. AB - Between September 1986 and June 1997, 24 children with high-risk ALL in CR1 were allografted after TAM (fractionated TBI, high-dose Ara-C, and melphalan; n = 10) or BAM protocol (busulfan, high-dose Ara-C, and melphalan; n = 14). The EFS for transplants from sibling donors was 33% with TAM and 62% with BAM (P = 0.148). The probability of acute GvHD was 70% with TAM and 15% with BAM (P = 0.003). Four of 17 evaluable patients relapsed: one after TAM and three after BAM. In all, 46 other children transplanted in CR beyond CR1 were studied for sequelae. Long-term side effects were more frequent in TAM vs BAM. In children with ALL, busulfan may be a good alternative to TBI to improve the quality of life. PMID- 15354204 TI - Quality of life assessment in patients undergoing reduced intensity conditioning allogeneic as compared to autologous transplantation: results of a prospective study. AB - The aim was to analyze quality-of-life (QOL) during the first year post transplant in 47 patients undergoing reduced-intensity conditioning (RIC) allotransplantation, and to compare these with a similar subgroup of patients receiving autologous stem cell transplantation (ASCT). We used self-reported questionnaires. Each answer scored from 0 (not at all) to 4 (very much), with higher scores indicating worse functioning. Mean value of physical categories among RIC transplants ranged between 1.23 and 0.77 indicating that patients scored very low for physical symptoms. Patients undergoing ASCT had higher scores in questionnaires performed early after transplant and then gradually improved (P < 0.001). Overall, when we compared physical functioning scores, allo-RIC did significantly better (P = 0.049). Nevertheless, while allo-RIC scores were significantly better for the first three questionnaires, ASCT patients did better in the last two questionnaires. These findings are in accordance with the toxicities observed in both subgroups which are lower in the RIC group early after transplant. No significant differences were observed between either subgroup for any of the functional, social/ family, psychological distress and satisfaction with doctor/nurse relationship items. We have observed similar QOL among patients undergoing RIC-allo as compared to ASCT although GVHD remains an important 'event' in QOL. PMID- 15354205 TI - Outcome of allogeneic stem cell transplantation in patients with myelofibrosis. AB - Myelofibrosis, either de novo or following pre-existing hematologic diseases, can be cured by allogeneic hematopoietic stem cell transplantation (SCT), but SCT is associated with significant morbidity and mortality, making the choice and timing of transplantation difficult. In all, 20 patients (seven female and 13 male), with a median age of 45 years (range 22-57 years), with idiopathic myelofibrosis (n = 12), post-thrombocythemic (n = 3) or post-polycythemic (n = 2) myeloid metaplasia or leukemic transformation (n = 3), underwent allogeneic SCT at our center between 1994 and 2003. With regard to the pre-transplant presence of risk factors such as hemoglobin levels < or =10 mg/dl, grade III marrow fibrosis or peripheral blast counts >1%, patients were divided into high- and low-risk groups. The estimated 3-year survival post transplant was 38.5% for all patients. The 3-year probability of survival within the high-risk group (n = 11) characterized by the presence of at least two risk factors was 16%. Low-risk patients (n = 9) with at most one risk factor had an estimated 3-year survival of 67%. Thus, previously defined risk determinants for the outcome of allogeneic transplantation for myelofibrosis may provide useful information facilitating treatment strategies. Our data suggest that transplantation should be taken into consideration before poor prognostic variables develop. PMID- 15354206 TI - High-dose thiotepa, melphalan and carboplatin (TMCb) followed by autologous peripheral blood stem cell transplantation in patients with lymphoma -- a retrospective evaluation. AB - The purpose of this evaluation was to investigate the efficacy of high-dose chemotherapy with thiotepa, melphalan, and carboplatin (TMCb), and of autologous peripheral blood stem cell (PBSC) infusion in patients with aggressive non Hodgkin's lymphoma (NHL) or Hodgkin's disease (HD). A total of 42 patients, 23 with intermediate-grade NHL and 19 with HD, received thiotepa (500 mg/m2), melphalan (100 mg/m2), and carboplatin (1050-1350 mg/m2) followed by autologous PBSC infusion. Of 21 patients with more advanced disease, four had primary refractory disease, one was in complete remission (CR)-2, 11 were in first refractory relapse, and five were beyond first relapse. Of 21 patients with less advanced disease, two were in CR-1, four were in CR-2, and 15 were in first responding relapse. In all, 14 patients (33%) had received prior radiotherapy prohibiting a total-body irradiation (TBI)-based conditioning regimen. The projected 2-year probabilities of survival, event-free survival (EFS), and relapse for all patients were 0.65, 0.60, and 0.21 (0.85, 0.80, and 0.10 for patients with less advanced disease and 0.47, 0.42, and 0.33 for patients with more advanced disease). The probability of nonrelapse mortality in the first 100 days was 0.12. Grade 3-4 regimen-related toxicities (RRT) occurred in five of 42 (12%) patients and death due to grade-4 RRT occurred in only one (2.5%) patient. These preliminary data suggest that 0.42% EFS in this study for advanced disease patients is highly encouraging and high-dose TMCb followed by autologous PBSC transplantation is well tolerated as well as an effective regimen in patients with intermediate-grade NHL or HD, and may be comparable to some previously used regimens including TBI-based regimens. PMID- 15354207 TI - Favorable outcome of allogeneic hematopoietic stem cell transplantation for relapsed or refractory acute promyelocytic leukemia in childhood. AB - The optimal therapy for children with relapsed or refractory acute promyelocytic leukemia (APL) is unclear. We therefore reviewed our institutional outcomes for children undergoing allogeneic hematopoietic stem cell transplantation (HSCT) for advanced APL. Between 1986 and 2003, 12 allogeneic HSCTs (five related donor, seven unrelated donor) were performed for 11 patients (median age, 13 years) with relapsed (n = 8) or refractory (n = 3) APL. All patients engrafted, after a median of 18.5 days. Grade B-D acute graft-versus-host disease (GVHD) developed after five transplants (42%; 90% CI, 18-68%), and the cumulative incidence of chronic GVHD was 45% (90% CI, 19-71%). The cumulative incidence of overt relapse post-HSCT was 10% (90% CI, 0-28%). The overall 5-year survival was 73% (90% confidence interval (CI), 51-95%), with a median post-HSCT follow-up of 64 months. The Lansky/Karnofsky performance scores are 100% in six of eight survivors. In view of the low risk of subsequent relapse and favorable survival suggested by other reports and our own experience, we continue to recommend allogeneic HSCT for children with advanced APL for whom a suitably HLA-matched donor is identified. PMID- 15354208 TI - T-cell post-transplant lymphoproliferative disorder after hematopoietic stem cell transplantation: another case and a review of the literature. PMID- 15354209 TI - Phase II trial of the antiangiogenic agent IM862 in metastatic renal cell carcinoma. AB - IM862 is a naturally occurring dipeptide (L-glu-L-trp) with immunomodulatory and antiangiogenic properties. A significant anticancer activity has been reported recently in AIDS-related Kaposi's sarcoma, a tumour of endothelial cell origin. The high vascularity and responsiveness to immunotherapy of renal cell carcinoma (RCC) makes such a tumour a potential target for IM862. In all, 25 patients were accrued in a prospective phase II trial using a standard two-step design. The main inclusion criteria were WHO performance status 3 months, normal marrow, kidney and liver functions. IM862 was given intranasally at a dose of 20 mg three times daily. Each cycle consisted of 8 consecutive weeks of treatment. All 25 patients were fully evaluable for response and 24 for toxicities. Median age was 62 years (range 42-76), median WHO PS was 1 (0-2). No grade 2 or 3 toxicities related to the study drug have been recorded. Eight patients had stable disease (SD) and 17 progressed while on treatment. Median survival was 7.9 months (range 2.7-20). Median time to progression was 1.9 months (range 1.2-12.6). Median duration of SD was 6 months (range 5.2-12.6+). Analysis of blood angiogenic markers showed a significant decrease of plasma vascular endothelial growth factor (VEGF) levels after 4 and 8 weeks of therapy. Treatment with IM862 has no toxicity, but does not lead to any significant objective responses in metastatic RCC. IM862 should not be further evaluated as a single agent at these doses and schedule for this population of patients. The decrease in VEGF levels warrants further investigation of IM862 as an antiangiogenic therapy. PMID- 15354210 TI - MSH6 missense mutations are often associated with no or low cancer susceptibility. AB - Mismatch repair (MMR) deficiency in tumours from patients with the hereditary nonpolyposis colorectal cancer (HNPCC) syndrome is mainly caused by mutations in the MLH1, MSH2, and MSH6 genes. A major challenge in the clinical management of patients with suspected HNPCC is the frequent occurrence of missense mutations in MSH6. These can be considered neither deleterious nor clinically innocent a priori. To assess their significance we studied five novel MSH6 missense mutations in six patients derived from a series of consecutive endometrial and colorectal cancer patients selected for study after their tumours were determined to be microsatellite unstable. We tested each mutated protein for heterodimerisation with MSH2 and for in vitro MMR capability. Four mutations (R128L, P623L, K728T, G881K+S) showed no impairment of these functions while the fifth (E1193K) displayed marked impairment of both functions. These results, taken together with our previous similar findings concerning six other missense mutations in MSH6, allow us to conclude that many or most missense changes in MSH6 likely are clinically innocent, whereas some missense changes such as E1193K, which lead to impaired MMR, are likely to be clinically significant, but have low penetrance. PMID- 15354211 TI - Latent effects of fibronectin, alpha5beta1 integrin, alphaVbeta5 integrin and the cytoskeleton regulate pancreatic carcinoma cell IL-8 secretion. AB - Interactions between tumour cells and the microenvironment are increasingly recognised to have an influence on cancer progression. In pancreatic carcinoma, a highly desmoplastic stroma with abnormal extracellular matrix (ECM) protein and interleukin-8 (IL-8) expression is seen. To investigate whether the ECM may further contribute to abnormalities in the microenvironment by influencing IL-8 secretion, we cultured the Mia PaCa2 pancreatic carcinoma cell line on fibronectin. This resulted in a dose-dependent increase in IL-8 secretion, which was RGD dependent and accompanied by cell spreading and proliferation. The role of spreading was assessed by disruption of the cytoskeleton with cytochalasin D, resulting in a large increase in IL-8 secretion, which was reduced from 31- to 24 fold by fibronectin. This remarkable response was associated with inhibition of spreading and proliferation and represents a novel cytoskeletal function. To investigate whether it could be accounted for by the loss of integrin-mediated signalling, the expressed alpha5beta1, alphaVbeta5 and alpha3beta1 integrins were inhibited. alpha5beta1 inhibition prevented spreading and proliferation but produced a much smaller rise in IL-8 secretion than cytochalasin D. alphaVbeta5 inhibition alone had only minor effects but when inhibited in combination with alpha5beta1 completely abolished the response to fibronectin. These results reveal latent stimulatory effects of the alphaVbeta5 integrin on IL-8 secretion and suggest that integrin crosstalk may limit the induction of IL-8 secretion by fibronectin. However, the magnitude of IL-8 secretion induced by cytochalasin cannot be accounted for by integrin signalling and may reflect the influence of another signalling pathway or a novel, intrinsic cytoskeletal function. PMID- 15354212 TI - Constitutional activation of IL-6-mediated JAK/STAT pathway through hypermethylation of SOCS-1 in human gastric cancer cell line. AB - The interleukin-mediated Janus kinase (JAK)/STAT pathway plays a crucial role in carcinogenesis. Recently, increased STAT3 activity was found in hepatocellular carcinoma and multiple myeloma in which there was silencing of SOCS-1 (suppressor of cytokine signalling-1) by gene promoter hypermethylation. We investigated the expression level of interleukin-6 (IL-6) and SOCS-1 in gastric cancer cell lines. Expression of SOCS-1 correlated with IL-6 level in most of the cell lines, except for AGS cells in which SOCS-1 was absent despite a high level of IL-6 production. Methylation analysis by methylation-specific polymerase chain reaction and bisulphite sequencing revealed that CpG island of SOCS-1 was densely methylated in AGS cells. Demethylation treatment by 5'aza-deoxycytidine restored SOCS-1 expression and also suppressed constitutive STAT3 phosphorylation in AGS cells. Moreover, methylation of SOCS-1 was detected in 27.5% (11 of 40) of primary gastric tumours samples, 10% (one of 10) of adjacent noncancer tissues but not in any (zero of nine) normal gastric mucosa. Methylation of SOCS-1 also correlated with the loss of mRNA expression in some primary gastric cancers. In conclusion, this is the first report to demonstrate that hypermethylation of SOCS-1 led to gene silencing in gastric cancer cell line and primary tumour samples. Downregulation of SOCS-1 cooperates with IL-6 in the activation of JAK/STAT pathway in gastric cancer. PMID- 15354213 TI - Bax regulates c-Myc-induced mammary tumour apoptosis but not proliferation in MMTV-c-myc transgenic mice. AB - The expression of the proto-oncogene c-myc is frequently deregulated, via multiple mechanisms, in human breast cancers. Deregulated expression of c-myc contributes to mammary epithelial cell transformation and is causally involved in mammary tumorigenesis in MMTV-c-myc transgenic mice. c-Myc is known to promote cellular proliferation, apoptosis, genomic instability and tumorigenesis in several distinct tissues, both in vivo and in vitro. Expression of the proapoptotic regulatory gene bax is reduced or absent in human breast cancers, and c-Myc has been shown to regulate the expression of Bax, as well as cooperate with Bax in controlling apoptosis in a fibroblast model. Additionally, loss of bax reduces c-Myc-induced apoptosis in lymphoid cells and increases c-Myc mediated lymphomagenesis in vivo. In order to assess whether loss of bax could influence c-Myc-induced apoptosis and tumorigenesis in the mammary gland in vivo, we generated MMTV-c-myc transgenic mice in which neither, one, or both wild-type alleles of bax were eliminated. Haploid loss of bax in MMTV-c-myc transgenic mice resulted in significantly reduced mammary tumour apoptosis. As anticipated for an apoptosis-regulatory gene, loss of the wild-type bax alleles did not significantly alter cellular proliferation in either mammary adenocarcinomas or dysplastic mammary tissues. However, in contrast to c-Myc-mediated lymphomagenesis, loss of one or both alleles of bax in MMTV-c-myc transgenic mice did not significantly enhance mammary tumorigenesis, despite evidence that haploid loss of bax might modestly increase mammary tumour multiplicity. Our results demonstrate that Bax contributes significantly to c-Myc-induced apoptosis in mammary tumours. In addition, they suggest that in contrast to c-Myc-induced lymphomagenesis, mammary tumorigenesis induced by deregulated c-myc expression requires some amount of Bax expression. PMID- 15354214 TI - Zinc-finger protein ZNF165 is a novel cancer-testis antigen capable of eliciting antibody response in hepatocellular carcinoma patients. AB - ZNF165 is a zinc-finger protein gene that was identified from human adult testis. Analysis of the origins of publicly available expressed sequence tags as presented in Unigene and SAGE databases revealed that ZNF165 mRNA was expressed in tumours of different tissues. RT-PCR, real-time PCR and Northern blotting analysis confirmed that ZNF165 mRNA was expressed in the hepatocellular carcinoma, gastric cancer, colon cancer and non-small-cell lung carcinoma. The nucleotide sequence of ZNF165 expressed in tumours is identical to that expressed in the testis. Humoral responses of hepatocellular carcinoma (HCC) patients against ZNF165 protein were determined by Western blotting using the recombinant ZNF165 protein. Antibodies against ZNF165 protein were detected in approximately 5% (four of 82) of the sera from HCC patients. These results suggest that ZNF165, a member of the ZNF family, is a novel CT antigen capable of eliciting humoral immune response and be involved in tumour biology. PMID- 15354215 TI - Thymidylate synthase predictive power is overcome by irinotecan combination therapy with S-1 for gastric cancer. AB - The predictive values of thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) gene expressions were retrospectively evaluated in patients with gastric cancer treated by a regimen containing S-1. The study population consisted of 53 patients registered into different two phase II studies for metastatic gastric cancer; 27 patients treated by S-1-alone study: 26 patients treated with S-1 combined with irinotecan (CPT-11). TS and DPD gene expressions in primary tumours were measured by the real-time reverse transcription PCR method. There was no statistical difference in DPD gene expression in terms of response in cases treated with S-1 alone and those treated with S-1 plus CPT-11. TS mRNA of responding tumours was lower than that of nonresponding ones when treated with S-1 (P<0.005). In the S-1-alone group, taking TS cutoff as the median values, the response rate in the low TS group was 50%, but only 8% in the high TS group (P<0.05). Patients with low TS gene expression survived longer than those with high TS gene expression (P<0.0001). However, there was no statistically significant difference in response rate and survival between patients with low TS tumours and those with high TS tumours, when the cutoff was taken as the median value of TS gene expression in the group treated with S-1 plus CPT-11. In conclusion, treatment effects of S-1 monotherapy for gastric cancer were determined by the status of TS gene expression, regardless of DPD gene expression. TS predictive power was overcome by CPT-11 combination therapy with S-1. PMID- 15354216 TI - Balancing false positives and false negatives for the detection of differential expression in malignancies. AB - A basic problem of microarray data analysis is to identify genes whose expression is affected by the distinction between malignancies with different properties. These genes are said to be differentially expressed. Differential expression can be detected by selecting the genes with P-values (derived using an appropriate hypothesis test) below a certain rejection level. This selection, however, is not possible without accepting some false positives and negatives since the two sets of P-values, associated with the genes whose expression is and is not affected by the distinction between the different malignancies, overlap. We describe a procedure for the study of differential expression in microarray data based on receiver-operating characteristic curves. This approach can be useful to select a rejection level that balances the number of false positives and negatives and to assess the degree of overlap between the two sets of P-values. Since this degree of overlap characterises the balance that can be reached between the number of false positives and negatives, this quantity can be seen as a quality measure of microarray data with respect to the detection of differential expression. As an example, we apply our method to data sets studying acute leukaemia. PMID- 15354217 TI - Expression of the transcription factor CTCF in invasive breast cancer: a candidate gene located at 16q22.1. AB - CTCF is a ubiquitous 11-zinc-finger protein that plays a role in gene silencing or activation, chromatin insulation and genomic imprinting. The CTCF gene has been mapped to the chromosome band 16q22.1 that shows frequent loss of heterozygosity in breast cancer. The E-cadherin gene is the known tumour suppressor gene (TSG) at this region in lobular carcinomas; however, the target gene in the more frequent ductal tumours is still unknown. Since CTCF targets include TSGs and oncogenes and it has the ability to inhibit cell growth and proliferation, it has been suggested that it may be the target gene at the 16q22.1 in ductal carcinomas. In the present study, tissue microarray technology was used to study the expression pattern of CTCF immunohistochemically in 344 cases of invasive breast carcinoma and its expression was correlated with clinicopathological variables and patient outcome. Results showed that breast tissues express CTCF in the parenchymal cells of the normal ducts and lobules but with a variable percentage of positive cells. Staining of CTCF was detected in the nuclei and cytoplasm of the malignant cells, but no significant loss or decrease of expression was noticed in association with any specific tumour type. There was a significant correlation between expression of CTCF and histological grades; lower expression was associated with grade 3 tumours. Cytoplasmic expression was associated with increased tumour size and with the presence of vascular invasion. However, no association was found between CTCF expression and tumour type, lymph node stage, oestrogen receptor expression or patient outcome. In conclusion, the current results show that CTCF, although it may play a role in breast carcinogenesis, is unlikely to be the TSG targeted by the 16q22.1 loss in breast cancer and thus another gene or genes at this region remain to be identified. PMID- 15354218 TI - Gastric and intestinal phenotypic marker expression in gastric carcinomas and recurrence pattern after surgery-immunohistochemical analysis of 213 lesions. AB - Both gastric and intestinal phenotypic markers are known to be expressed in gastric carcinomas, irrespective of their histologic type. In the present study, the relation between gastric and intestinal phenotypic marker expression in gastric carcinomas and the recurrence pattern after surgery was examined. The phenotypic marker expression of the tumour was determined by examining the expression of human gastric mucin (HGM), MUC6, MUC2 and CD10 in 213 advanced gastric carcinomas in 213 patients who had undergone a curative resection (97 died from recurrent gastric carcinoma and 116 were alive without recurrence at the end of the follow-up period). Tumours were classified into gastric (G), gastric and intestinal mixed (GI), intestinal (I) or unclassified (UC) phenotypes according to the immunopositivity of HGM, MUC6, MUC2 and CD10 stainings. The incidence of HGM-positive tumours and MUC2-negative tumours was significantly higher in tumours with peritoneal recurrence than in tumours without recurrence (73.3%, 44 out of 60 cases vs 54.3%, 63 out of 116 (P=0.022); and 70.0%, 42 out of 60 vs 38.8%, 45 out of 116 (P=0.0002), respectively). The incidence of G phenotype tumours was also significantly higher in tumours with peritoneal recurrence than in tumours without recurrence (58.3%, 35 out of 60 cases vs 28.4%, 33 out of 116 (P=0.0002)). The incidence of MUC2-negative tumours and CD10 positive tumours was significantly higher in tumours with haematogenous recurrence than in tumours without recurrence (62.5%, 20 out of 32 cases vs 38.8%, 45 out of 116 (P=0.028); and 43.8%, 14 out of 32 vs 23.3%, 27 out of 116 (P=0.039); respectively). Our present findings show that the gastric and intestinal phenotypic marker expression of the tumour, determined by immunohistochemical staining for HGM, MUC6, MUC2 and CD10, can be used to predict the pattern of gastric carcinoma recurrence after curative resection. PMID- 15354219 TI - The effect of cigarette smoking use and cessation on serum insulin-like growth factors. AB - The patterns of risk association between circulating levels of insulin-like growth factor (IGF)-I, and its main binding protein, IGFBP-3, differ between smoking and nonsmoking-related cancers. To investigate this observation further, we measured serum IGF-I, IGF-II and IGF-binding protein-3 concentrations in 232 men and 210 women (aged 55-64 years), and related peptide levels to smoking characteristics. Current smoking was associated with significant reductions in mean IGFBP-3 levels in men assessed by the number of cigarettes smoked daily (P(trend)=0.007) and pack-years smoked (P(trend)=0.03). Mean IGF-I levels decreased with increasing cigarette use in men (P(trend)=0.11). There were no patterns of association between smoking and IGF peptides in women. For male former vs never smokers, there were no differences in mean IGF-I and IGFBP-3 concentrations, suggesting that smoking cessation is associated with normalisation of peptide concentrations. PMID- 15354220 TI - The systemic inflammatory response, performance status and survival in patients undergoing alpha-interferon treatment for advanced renal cancer. AB - The prognostic value of C-reactive protein, compared with ECOG performance status (ECOG-ps), in patients receiving alpha-interferon treatment for advanced renal cancer was assessed in 58 patients. In all, 55 patients died on follow-up. On multivariate analysis with ECOG-ps and C-reactive protein entered as covariates, only C-reactive protein was a significant independent predictor of survival (HR 2.03, 95% CI 1.09-3.80, P=0.026). PMID- 15354221 TI - Extracellular proteases as targets for treatment of cancer metastases. AB - Metastasis, the dissemination of tumor cells to distant organs, is often associated with fatal outcome in cancer patients. Formation of metastasis requires degradation of extracellular matrices and several families of proteases have been implicated in this process, including matrix metalloproteinases (MMPs), serine and cysteine proteases. Inhibition of these enzymes in animal models of metastasis has shown impressive therapeutic effects. This report discusses the various approaches used for enzyme inhibition and describes new developments in drug design for inhibition of proteases in metastatic disease. PMID- 15354222 TI - Metal-Salen Schiff base complexes in catalysis: practical aspects. AB - Schiff base ligands are considered "privileged ligands" because they are easily prepared by the condensation between aldehydes and imines. Stereogenic centres or other elements of chirality (planes, axes) can be introduced in the synthetic design. Schiff base ligands are able to coordinate many different metals, and to stabilize them in various oxidation states, enabling the use of Schiff base metal complexes for a large variety of useful catalytic transformations. Practical guidelines for the preparation and use of different Schiff base metal complexes in the field of catalytic transformations are discussed in this tutorial review. PMID- 15354223 TI - The synthesis of peptides and proteins containing non-natural amino acids. AB - Methods for the incorporation of non-natural amino acids into proteins have advanced significantly over recent years and in this tutorial review we aim to give a general overview of the area. These techniques offer the possibility of modulating the structures and functions of proteins and thus permit the generation of novel designed systems for both biocatalytic and mechanistic studies. Four complementary approaches are discussed in detail along with examples of their application. The advantages and disadvantages of each technique are also discussed. PMID- 15354224 TI - Non-stabilized transition metal carbenes as intermediates in intramolecular reactions of alkynes with alkenes. AB - In this tutorial review we summarize the two major pathways followed in the reaction of alkenes with alkynes catalysed by electrophilic transition metals. If the metal coordinates simultaneously to the alkyne and the alkene, an oxidative cyclometallation can ensue to give a metallacyclopentene, which usually evolves by [small beta]-hydrogen elimination to give Alder-ene cycloisomerisation derivatives. On the other hand, coordination of the metal to the alkyne promotes the attack of the alkene to give metal cyclopropyl carbenes. PMID- 15354225 TI - Unique asymmetric catalysis of cis-beta metal complexes of salen and its related Schiff-base ligands. AB - Complexes of chiral salen and its related tetradentate Schiff-base ligands adopt three different configurations, trans, cis-alpha and cis-beta. Of these complexes, trans-complexes have been widely used as catalysts for various asymmetric reactions. However, recent studies have disclosed that cis-beta metallosalen and its related complexes show unique asymmetric catalyses that cannot be achieved by trans-metallosalen complexes. The present article summarizes generation of cis-beta metallosalen and its related complexes, their structural features, and their application to asymmetric syntheses. PMID- 15354226 TI - The role of coordination chemistry in the development of target-specific radiopharmaceuticals. AB - There is a great current interest in developing target-specific radiopharmaceuticals for early detection of diseases and radiotherapy of cancers. This critical review will focus on the role of coordination chemistry in the development of target-specific radiopharmaceuticals. It will also discuss the recent development in technetium, copper, gallium, indium, yttrium and lanthanide chemistry, as well as analytical tools for quality control and characterization of radiolabeled small biomolecules (159 references). PMID- 15354227 TI - [Vagus nerve stimulation in drug-resistant epilepsy. Experience with 23 patients]. AB - BACKGROUND: Vagal nerve stimulation (VNS) is a new non-pharmacological method of pharmacoresistant epilepsy treatment. The aim of this paper was to present effects of treatment in 23 patients with drug-resistant epilepsy with a different etiology. MATERIAL AND METHODS: Implantation and treatment was performed in two centers in 1998-2002. The effect of treatment was presented as a reduction of seizures during therapy. RESULTS: The lack of group homogeneity and a small number of patients (especially a small number of patients with a long follow-up period) did not allow a more detailed analysis to be made, although there seems to be a clear tendency to obtain better effects of treatment over follow-up time (at 24 month more than 50% seizure reduction or cease of seizures was observed in 80% of patients). The possibility to turn on the device "on demand" is an important advantage of this method. This raises the effectiveness of treatment in more than 80% of patients, and in more than 20% it stops the seizure. There were two groups of undesired side effects: frequent specific effects caused by local irritation of the vagal nerve in the cervical part of the neck and rare transient general effects. Both groups of effects rarely caused any treatment complications. CONCLUSIONS: VNS is an effective method of treatment, complementary to other epilepsy treatment methods and should be used in patients with drug-resistant epilepsy as an alternative to neurosurgical treatment. VNS improves the quality of life in treated patients. PMID- 15354229 TI - [Surgical treatment of intraventricular tumors]. AB - In the last 11 years we operated on 20 patients (1.03% of all operated brain tumors) with lateral ventricular masses. Anamnesis lasted from 6 months in the case of epileptic seizures to 6 years when headache was the main symptom of disease only. Tumor was located in the anterior cornu and pars centralis in 15, in the trigonum of lateral ventricle in 2, and in the posterior cornu of the lateral ventricle in 3 cases. 13 tumors were large and in MRI the largest diameter was above 5 cm. Tumors located in the anterior cornu and pars centralis were removed using transcortical (10 cases) or transcallosal approaches (5 times). The tumor of the trigonum of ventricle was removed twice through superior parietal lobulus. Tumors of the posterior cornu were operated on via precuneus in order to save visual fields or through the occipital cortex. In 4 cases after removal of tumor the orifice in the frontal lobe was closed with fibrine glue. We totally removed tumors from lateral ventricles in 85% and subtotally in 15% of all cases. Histopathological examination revealed: astrocytoma anaplasticum in 5 cases, astrocytoma pilocyticum 4, oligodendroglioma 2, oligodendroglioma mixtum 1, papilloma plexus chorioidei 2, cystis epidermalis 2, subependymoma 1, ependymoma 1, pineocytoma 1 and haemangiona arterio-venosum 1. Two patients (10%) with diagnosis of anaplastic astrocytoma died after the operation as a result of ischemia and edema cerebri. During the follow-up examination, where the average time of observation was 3.7 years, 15 patients were found to be in a good condition (GR+MD), 1 patient was severely disabled (SD), 2 patients with anaplastic astrocytoma and pineocytoma died because of enlargement of the remnants of tumors left at first operation in spite of reoperation. Tumors of the lateral ventricle grow slowly and often achieve a large size before their clinical appearance. Most of tumors are accessible for a total or subtotal removal with acceptable morbidity and mortality rate after the operation. Long term results depend on the size of resection and histopathological investigation. PMID- 15354230 TI - [Importance of early operative decompression of spinal cord after cervical spine injuries]. AB - BACKGROUND AND PURPOSE: There is no standard timing of the spinal cord decompression. Experimental animal models and clinical investigations on Methylprednisolone (NASCIS-2 and -3) indicate that the time up to 8 hours is the optimal therapeutic window for the early spinal cord decompression. We accepted this time window in our practice. A retrospective clinical evaluation of the early (up to 8 hours) operative decompression of the injured cervical spinal cord was undertaken. MATERIAL AND METHODS: The early operative decompression (range of 2-8 hours) of the cervical spinal cord was done in 32 patients (82% of operated cervical spinal cord injured patients). The neurologic clinical status of patients was assessed according to the ASIA impairment scale. In neurological terms 7 patients were completely and 25 incompletely impaired. Cervical spine injuries included 26 fractures and fracture-dislocations and the rest (6 cases) involved dislocations and disc ruptures. The mean patient age was 31.2 years (range of 16-69) and the average follow-up time was 13 months (range of 6-24). Methylprednisolone standard treatment was applied in all patients on admission to hospital. After diagnostic examinations patients were qualified and immediately operated. We performed 25 corpectomies, 6 discectomies and 1 decompressive laminectomy with stabilization in all cases. RESULTS: The final assessment covers 29 patients because 3 (9.4%) of them died during 4-6 weeks after operation. One patient has not improved and still had A degree but 28 remaining patients (94.5%) have improved neurologically at follow-up. 69% of them recovered by one degree and 27.6% by two degrees on the ASIA scale. In the early postoperative stage during their hospital stay 47% of patients had one degree improvement. CONCLUSIONS: The early cervical spinal cord decompression creates optimal conditions for the neurological recovery. Encouraging results presented in the paper indicate the purposefulness of the decompression performed as fast as possible and indicate the necessity to make emergency services more efficient in order to shorten delivery time to specialized spine centers. PMID- 15354231 TI - [Optimal use of the lumbar infusion test in patients with communicating hydrocephalus. Theoretical considerations in comparison to clinical observations]. AB - BACKGROUND AND PURPOSE: Lumbar infusion test (IT) is still one of the most important diagnostic tools in the communicating hydrocephalus. On one hand suitability of parameters achieved during tests is being highlighted, while on the other hand benefits are being questioned due to the test invasiveness. The aim of this paper is to assess the possibility of reducing the duration of IT. The study was based on a theoretical model and the analysis of selected IT performed in patients suffering from normal pressure hydrocephalus. MATERIAL AND METHODS: Theoretical principles of the volume-pressure compensation in hydrocephalic patients are considered. Then, based on selected infusion test, an analysis of the data was carried out using the mathematical model. Finally the methods of IT optimization were proposed. RESULTS: The analysis of IT should be performed in dynamic (not static) conditions. For the proper assessment of the IT the classical Marmarou's model is sufficient. This model interposed on the normal IT curve helps avoid too short and too long time of the IT. From a theoretical point of view it is impossible to minimize the duration of the IT by changing the infusion rate. This is due to the fact that formal relationships are characterized by curves with a single maximum, but not a minimum. The proposed algorithm for predicting the level of the steady state pressure, based on the initial course of the IT curve, allows for about 30% reduction of the IT time. The predictive error for the pressure value is less than 10%. PMID- 15354232 TI - [Changes in lipoprotein (a) [Lp(a)] level after an ischemic stroke]. AB - The aim of the work was to recognize whether often observed high levels of apolipoprotein (a) [Lp(a)] in patients shortly after an ischemic stroke are a result of the acute phase reaction. In 13 patients Lp(a) was determined within the first 24 hours after the stroke onset, after the next 7 days and after three months i.e. when it could be considered that Lp(a) level was the same as before onset of the disease. In 17 patients only two determinations were performed. Another acute phase indicator: C-reactive protein (CRP), as well as serum lipids were also determined. CRP level was increased in the first determination and increased further after 7 days. After three months it returned to low values. High density lipoprotein (HDL) cholesterol which demonstrates a negative acute phase response changed in the opposite way. No similar fluctuations of Lp(a) level were observed. It can be concluded that during the investigated period Lp(a) had no properties of the acute phase reactant. PMID- 15354233 TI - [5-Aminolevulinic acid (ALA) and its applications in neurosurgery]. AB - Delta-aminolevulinic acid (ALA) is a precursor of the synthesis of porphyrins including heme produced in all mammalian cells. Exogenous ALA induces selective accumulation of the other heme precursor, protoporphyrin IX (PpIX), in neoplastic cells, such as those of malignant gliomas. Upon exposure to violet-blue light PpIX becomes activated, which results in red-light fluorescence as well, as in photodynamic oxidations which may be lethal to the cells. In neurosurgery ALA is used for intraoperative labeling of the border regions of malignant gliomas infiltrated by alive clonogenic tumor cells (ALA-PDD), and is helpful in precise resection of these regions. Clinical data indicate that ALA-PDD-assisted resection of malignant gliomas may result in statistically significant prolongation of postoperative survival. Ongoing research concentrates also on the use of ALA for a selective elimination of glioma cells in situ, and on lipophilic ALA derivatives with more favorable pharmacokinetic properties. PMID- 15354234 TI - [Atherosclerotic plaque instability and ischemic stroke: the role of inflammatory and immunologic factors. Practical implications]. AB - The majority of strokes (85-90%) result from cerebral ischemia. In most cases extra - and intracranial vessel atherosclerotic changes are considered to be responsible for cerebral ischemia. A sudden failure of cerebral circulation is usually combined with instability of atherosclerotic plaques. Pathologic studies demonstrate that atherosclerosis simultaneously involves the whole arterial vessel tree. The mechanism of atherosclerotic plaque formation is similar in different regions of the arterial system, including the brain supplying arteries. The essence of atherosclerosis appears to be an excessive inflammatory fibroproliferative response to various forms of the arterial wall injury. The development of unstable plaques is closely related to the inflammatory process involving the arterial wall. Immunological factors seem to play an important role in the development of atherosclerotic changes and their destabilization. Imaging techniques for arteries supplying the brain are known. None of them gives a clear answer about plaque instability. Taking into account the role of the inflammatory process in the pathogenesis and instability of atherosclerotic changes, measurement of serologic markers of plaque instability seems to be complementary to imaging methods for diagnosis of unstable atherosclerotic plaques. The inflammatory aspect of atherosclerosis should be reflected in the prophylaxis of the central nervous system vascular disorders. PMID- 15354235 TI - [Spontaneous spinal cord herniation: a rare cause of the Brown-Sequard syndrome]. AB - Recent reports have shown that spontaneous spinal cord herniation in the thoracic segment of the spine may be one of the causes of the progressive spinal cord lesion. Although it was described for the first time by Wortzman in 1974, it was only in early 1990s that a growing number of publications on single cases of the condition started to be observed. In the relevant literature collected by us we found reports on 53 patients altogether. In a number of cases herniation was diagnosed only intraoperatively in spite of complex radiological diagnostics. The analysis of relevant literature shows that spontaneous or idiopathic spinal cord herniation has a very typical clinical picture and most cases share a number of features, such as the clinical condition, age and sex of the patient as well as the level and location of the entity and its radiological picture. We conclude that spontaneous herniation of the spinal cord may be not as rare as previously thought and that it should be always taken into account in differential diagnostics of progressive myelopathy in the thoracic segment in middle-aged patients, especially females. PMID- 15354236 TI - [Guillain-Barre syndrome with central nervous system symptoms. Report of two cases]. AB - Guillain-Barre syndrome - acute inflammatory demyelinating polyradiculoneuropathy - is characterized by symmetrical flaccid paresis of limbs and areflexia or hyporeflexia which progress over a few days, up to 4 weeks. The central nervous system lesion is rarely reported in the course or treatment of the disease. In the paper two cases of patients with diagnosed Guillain-Barre syndrome with the central nervous system manifestations were discussed. A case of a 55-year-old woman was presented, who during hospitalization, on the last day of intravenous immunoglobulin therapy developed a hallucinatory syndrome. Furthermore, a case of a 18-year-old female patient with classic features of Guillain-Barre syndrome was described, because of its atypical initial presentation (headache, drowsiness and meningismus). PMID- 15354237 TI - [Features of intracranial hypertension as a manifestation of cauda equina tumor]. AB - A 61-year-old man was admitted to the Department of Neurology, Medical University of Bialystok with signs of intracranial hypertension associated with headaches and papilledema, with no other signs of the nervous system dysfunction. CT and MRI brain scans were normal, cerebrospinal fluid contained 1620 mg/dl of proteins. One month later epileptic seizures with a loss of consciousness occurred. Brain CT and MRI scans showed no focal pathology and failed to disclose any intracranial tumor. The acute pain in the lumbar spine region made us perform an MRI scan of the lower spinal cord, which disclosed cauda equina tumor. The tumor was surgically removed and it was histopathologically concluded to be ependymoma. After the operation there were neither headaches nor seizures and we noticed an improvement in his vision acuity. In the postoperative course the patient suffered from urinary and bowel dysfunction. In this article we discuss a pathogenesis of intracranial hypertension occurring in spinal cord tumors and stress the need for a diagnosis of spinal cord lesions in patients with the elevated intracranial pressure. PMID- 15354238 TI - [Carbon cage used for anterior lumbar interbody fusion: a case report]. AB - Two cases of patients with lumbar discopathy who underwent anterior lumbar interbody fusion (ALIF) were described. The authors stress the advantages and disadvantages of this kind of stabilization. PMID- 15354240 TI - [Bilateral cerebellar hematoma after supratentorial glioma surgery]. AB - We present a case of bilateral hematoma in cerebellar hemispheres in a 30-year old man after surgical treatment of extensive left frontal glioma. 16 hours after surgery the patient lost consciousness. An immediate CT revealed hematoma in both cerebellar hemispheres. The hematoma was subsequently removed via bilateral suboccipital craniectomy. After the operation the clinical status of the patient gradually improved - he was discharged in a good general condition. In the presented case the hematoma developed presumably as a consequence of extensive cerebrospinal fluid (CSF) loss (670 ml) via postoperative wound drainage. The resulting cerebellar displacement caused strain of the draining veins, affecting blood outflow, and causing parenchymal hemorrhage. In order to prevent the complication, massive CSF loss during and after operation should be avoided. Careful monitoring of the patient's condition in the postoperative period, even if the general status is good, is important because only an immediate intervention may prevent the development of irreversible consequences of cerebellar hematoma formation. PMID- 15354244 TI - [Use of the "c-clamp" in the emergency treatment of unstable pelvic fractures]. AB - It was the objective of this investigation to describe the results after stabilisation of pelvic fractures with the pelvic clamp in polytraumatized patients with unstable pelvic ring fractures. PATIENTS: Between 1999 and 2001 11 polytraumatized patients with an average age of 38 years with unstable fractures of the pelvic ring were treated with the "pelvic clamp" (PC). METHOD: The patient's data were analysed retrospectively from the moment they were admitted to our department until 48 hours after the treatment with the pelvic clamp. The following data were observed: -- The time period until the pelvic clamp was placed. -- The mean blood pressure. -- The oxygenation level (PaO (2)/FiO (2) coefficient). -- The number of requested blood units. -- The time period until hemodynamic stabilisation took place. RESULTS: 8 patients survived their injuries. 5 of them were admitted primarily to our department (ISS 39.8, PTS 35), the other 3 were secondarily admitted to our department (ISS 48.3, PTS 39). 3 of 11 patients (27 %) died averaged within the first 45 minutes after admittance. They also were treated in other units before admittance to our department. 8 surviving patients showed -- a hemodynamic stabilisation 6 hours after the treatment with the PC. -- an increase of the mean blood pressure about 25 % 20 minutes after the treatment with the PC. -- a stabilisation of the oxygenation level 6 hours after the treatment with the PC. -- a decrease of the number of requested blood units 6 hours after treatment with the PC. CONCLUSION: Even if the number of patients who were treated is small, the study shows a positive trend in terms of stabilisation of the vital parameters after stabilisation of the pelvic fracture with the pelvic clamp. PMID- 15354245 TI - [Evidence-based evaluation of present guidelines for the treatment of tennis elbow -- a review]. AB - The guidelines of the German Orthopaedic Societies regarding the treatment of lateral elbow epicondylitis were analysed on the ground of recently published reviews or randomised placebo-controlled trials (RCT). For the acute phase, reviews or RCTs failed to show a clinical effect beyond placebo if follow-up was extended over 6 weeks. For the chronic phase a current Cochrane review failed to identify any controlled trial regarding surgical procedures during the last decades. Without an adequate control group, it is not possible to draw any meaningful conclusions about the value of this modality of treatment. Therefore surgery is not indicated before repetitive low-energy extracorporeal shock wave therapy (ESWT) has been applied. This novel treatment, under strictly standardized conditions, showed effects beyond placebo in independent randomised placebo-controlled trials for follow-up periods of 3 and 6 months. To date there exists no evidence-based therapeutic algorithm for the treatment of acute and chronic tennis elbow. Even medium-term effects should be regarded as either a placebo effect or natural regression to the mean. PMID- 15354246 TI - [Operative results of treatment of intraarticular calcaneus fractures with the "low contact plate"]. AB - Calcaneus fractures are a major cause of accident- related invalidity. Long-term results are unsatisfactory with various therapy concepts as reported in the literature. Objective of this study is the long-term clinical and radiological outcome of a patient group treated with a low contact plate. Subjective assessment of function by the patient was an important evaluation criterion. From 01/92 to 12/98 a total of 182 patients with 209 calcaneus fractures were treated. Out of them 95 patients with 109 calcaneus fractures had a follow-up over a period ranging from 6 to 88 months (average 37.9 months). Severe impairment of walking was reported by 34.7 % of the patients. Severe impairment for work was declared by 68.4 % of the patients. 74.7 % of the patients complained middle or severe burden-pain. 64.3 % of the patients felt their quality of life impaired. After the injury 75.8 % of the patients were still employed. Arthrosis of the inferior ankle joint was found in 91.5 % of the cases. Within this collective three patient groups were formed with different follow-up periods to evaluate the temporal course, showing no significant differences. Frequently good functional and subjective results cannot be expected even with standardized operative procedures. PMID- 15354247 TI - [Surgery in non-small cell lung cancer of the superior sulcus: results of a combined preoperative and postoperative irradiation regime]. AB - Due to local invasion of the chest wall, patients with non small cell carcinoma (NSCLC) of the superior sulcus have been treated mainly by the Paulson regime with radiotherapy followed by surgical resection. Recent published data on the use of concurrent irradiation and chemotherapy followed by surgical resection seemed very promising. The aim of the present study was to determine the value and benefit of a combined preoperative and postoperative radiotherapy regime (Sandwich irradiation), and which factors predict prognosis following resection. PATIENTS AND METHODS: Between 1986 and 2003, 64 patients with non-small cell carcinoma of the superior sulcus were managed in our department. 28 underwent surgical resection with combined preoperative 40 Gy and postoperative 20 Gy external beam radiotherapy. Time to death was calculated using the method of Kaplan and Meier. Survival after surgery was the end point of the study. The association of factors to end of life end points was analyzed using the log-rank test for univariate analysis. Median follow up was 13.1 months. RESULTS: The actuarial 5-year-survival for the overall population was 30.2 %. For surgically rendered complete resection (CR) patients with no mediastinal lymph node metastases, the 5-year-survival-rate was 53.2 %. The 30-day-mortality-rate was 0 %. Most significant prognostic factors were the mediastinal lymph node involvement and the stage of the disease. CONCLUSIONS: Results of this retrospective study show that patients with non-small cell carcinoma of the superior sulcus can experience a long-term survival which is well comparative to other patients with NSCLC. Surgical resection with a combined preoperative and postoperative radiotherapy regime is well accepted. Special care should be taken in patient selection to identify patients with advanced stage of the disease and mediastinal lymph node metastases. PMID- 15354248 TI - [Merendino procedure with preservation of the vagus for early carcinoma of the gastroesophageal junction]. AB - Optimal therapy for early carcinoma of the gastroesophageal junction remains uncertain. Treatment alternatives discussed today reach from endoluminal techniques to radical esophagectomy with 2- or 3-field lymphadenectomy. In this context, the Merendino procedure with preservation of the vagal innervation to the stomach appears as an interesting therapeutic alternative. This paper summarizes indications, operative technique, and functional results with respect to postoperative quality of life in view of 2 cases operated in our department. PMID- 15354249 TI - [Twist-guided thread control: a technique for faster and atraumatic continuous sutures]. AB - A simple technique by twisting the thread in clockwise or counter clockwise direction before tightening the suture can help to produce a regular vascular suture line. In a blinded experiment with 60 vascular anastomosis it was proved that the time needed for suture can be reduced by 15%, and that traumatic touch with surgical instruments can be diminished by 63%. PMID- 15354250 TI - [Analysis of the cost-income ratio for open and laparoscopic sigmoid resection]. AB - Based on the example of the sigmoid resection, we did a cost-income analysis evaluating the particular expenses and effects of the new DRG-based medical financing system. We analysed the cost of diagnostic tests, medication and nutrition, surgery, personnel and non medical expenses for 20 open and 27 laparoscopic operated patients. The cost of open surgery was calculated to be 3,288.44 euro per patient while laparoscopic surgery was calculated to cost 4,271.98 euro. Based on an average hospitalisation period of 16 days, the estimated income for the hospital was 5,738.98 euro. Independent of the length of hospitalisation, an income of 6,621.34 euro for the hospital was estimated using the new DRG-based medical financing system. Thus, implementation of the new DRG based medical financing system makes it possible to give patients the benefit of shorter hospital stays following laparoscopic surgery. The cost for the hospital, however, is only minimally reduced. PMID- 15354251 TI - [Endoscopic removal of large colorectal polyps]. AB - BACKGROUND: Due to the potential risk of malignancy and technical difficulties in achieving complete removal, large colorectal polyps represent a special problem for the endoscopist. The aim of this study was to evaluate the capabilities and risks of endoscopy in complete removal of large colorectal polyps. METHODS: Endoscopic polypectomy of 189 colorectal polyps (141 sessile, 48 pedunculated) larger than 3 cm in diameter (range 3-13 cm) was carried out. Sessile polyps were removed using the piecemeal technique. RESULTS: Histology showed an adenoma in 173 cases, and invasive carcinoma was present in the adenoma in 16 patients. Complete endoscopic removal was achieved in one session in 129 sessile polyps and all pedunculated polyps; the other patients required two to ten sessions. Bleeding occurred in 20 patients during polypectomy, and after polypectomy in four. Except for two cases, definitive hemostasis was immediately achieved by endoscopic treatment. Perforation occurred in four patients. 3 were treated conservatively, in one patient who showed an invasive carcinoma of the cecum, resection was performed. There was no mortality due to polypectomy. During a median follow-up period of 77 months (6-107 months), six patients presented with recurrence of a benign adenoma, which was treated endoscopically, and one patient presented with a recurrent invasive carcinoma, which was treated surgically. CONCLUSIONS: Endoscopic polypectomy is a safe and effective method of treating large colorectal polyps, associated with a low complication rate, reduced hospital stay and lower costs in comparison to surgical procedure. PMID- 15354252 TI - [International clinical recommendations on scar management]. AB - Many techniques for management of hypertrophic scars and keloids have been proven through extensive use, but few have been supported by prospective studies with adequate control groups. Several new therapies showed good results in small-scale trials, but these have not been repeated in larger trials with long-term follow up. This article reports a qualitative overview of the available clinical literature by an international panel of experts using standard methods of appraisal. The article provides evidence- based recommendations on prevention and treatment of abnormal scarring and, where studies are insufficient, consensus on best practice. The recommendations focus on the management of hypertrophic scars and keloids, and are internationally applicable in a range of clinical situations. These recommendations support a move to a more evidence-based approach in scar management. This approach highlights a primary role for silicon gel sheeting and intralesional corticosteroids in the management of a wide variety of abnormal scars. The authors concluded that these are the only treatments for which sufficient evidence exists to make evidence-based recommendations. A number of other therapies that are in common use have achieved acceptance by the authors as standard practice. However, it is highly desirable that many standard practice and new emerging therapies undergo large-scale studies with long-term follow-up before being recommended conclusively as alternative therapies for scar management. PMID- 15354253 TI - [Management of postoperative compartment syndrome -- case report]. AB - AIM: Differential diagnosis and management of the lower extremity compartment syndrome as a potentially devastating complication of prolonged surgery in the lithotomy position. CASE REPORT: A 55-year-old patient underwent radical cystoprostatovesiculourethrectomy including reconstruction of an ileal conduit because of a multifocal recurrent tumor of the urinary bladder (operating time > 8 hours). On the first postoperative day, the patient complained about swelling within the right calf leading to the suspicion of a deep vein thrombosis. Phlebography of the right leg revealed: i) thrombosis-untypical occlusion of the distal popliteal vein and ii) no detection of the deep vein within the right calf (femoral and iliac veins were with no pathological finding). Tissue pressure was as follows: right, 55 mmHg/left, 11 mmHg, underlining clinical suspicion of compartment syndrome. The patient underwent a fasciotomy of the right calf. Over the following 5 days, muscle edema decreased, allowing subsequent mobilization of the patient. On the 8 (th) postoperative day, the patient died unexpectedly due to an acute myocardial infarction. CONCLUSION: In case of a swelling of the lower extremity after long-lasting surgical interventions performed in lithotomy position, a compartment syndrome is one of the possible differential diagnoses, the consequences of which can be avoided by an early diagnostic and adequate treatment. PMID- 15354254 TI - [Restenosis of carotid artery after stenting associated bleeding in the vascular wall]. AB - Stenting is an alternative procedure to the endarterectomy in patients with severe (> 70 %) symptomatic stenosis of the carotid artery. Its value is actually tested in large controlled randomised multicenter studies. Many vascular surgeons postulate no benefits for the patients but a higher complication rate for the stenting procedure. Especially the possibility of mechanical irritations of the endothel caused by the stent itself seems to be a major problem. Despite thousands of stenting procedures of the carotid artery, there exist no reports of patients with direct mechanical irritation of the carotid artery through the stent itself. It is only published a higher risk of embolisation. This case of a 71-years old patient, who had a irradiation of the head and neck region 5 years ago for cancer therapy, is presented here. After the irradiation (60 gr) the stent was not so fixed in the carotid wall as usually. Therefore the stent caused a mechanical irritation of the sclerotic plaque with a bleeding in the carotid wall and subsequent severe stenosis at the end of the stent with higher level of infarction volume. This case demonstrates the risk of the theoretical postulated irritation of the endothel through the stent itself in a high movement region. PMID- 15354255 TI - [Esophageal perforation by portions of a wild boar]. AB - We report a case of esophageal perforation by impacted portions of a wild boar. The 77-year-old patient was admitted 5 days after an opulent dinner. He complained of dysphagia and retrosternal pain and showed signs of severe sepsis. Endoscopy revealed meat and bone portions impacted in the esophagus. Surprisingly, impactation had not happened near any of the anatomical narrowings, but in the middle third. A CT scan showed penetration of the foreign body into the mediastinum and purulent mediastinitis. The patient underwent transthoracic esophagectomy and drainage of the severe, purulent mediastinal infection. Postoperative multi organ failure required a prolonged treatment in the intensive care unit. After recovery the gastrointestinal continuity was restored with gastric pull-up in the anterior mediastinum. The further course was uneventful. PMID- 15354256 TI - [Intra-abdominal fibromatosis after appendectomy as cause for ileus]. AB - Aggressive intraabdominal fibromatosis is a rare response to surgical trauma. Characteristic is the local destroying growth. Only an operation in the purpose of R0-resection ends in cure. We report on a 29-year-old man, who developed intraabdominal fibromatosis as reaction to surgical trauma 19 years after appendectomy and subsequent to several relaparotomies due to adhesions. A huge conglomeratumor which included the ileum and the colon acsendens resulted in bowel obstruction and ileus. Complete surgical removal of the tumor lead after a primary complicated course to recovery. PMID- 15354257 TI - [Influence of non-selective and cyclooxygenase-II-selective cyclooxygenase inhibitors on primary human osteoblasts]. PMID- 15354258 TI - [Five cytostatic substances in animal studies for prevention and treatment of experimentally induced peritoneal carcinomatosis]. AB - High local recurrence rates within the previous tumor bed or at the peritoneum remain an unsolved problem after surgical resection of malignant gastrointestinal tumors such as gastric, colorectal or pancreatic carcinoma. Currently, there are no standardized treatment protocols available for the prevention or treatment of peritoneal carcinomatosis. In a basic experimental trial, mitomycin, cisplatin, 5 FU, oxaliplatin and CPT-11 were used to prevent or treat peritoneal carcinomatosis induced in rats. Experiments were performed in three groups (n = 8 each) of animals plus two control groups. In the first group, Mitomycin, Cisplatin, 5-FU, Oxaliplatin and CPT-11 (n = 24 each) were applied directly following tumor cell implantation into the peritoneal cavity. In the second group, early postoperative intraperitoneal (i. p.) chemotherapy (day [d] 5, 10, 15 following surgical intervention for tumor cell transfer) was administered, whereas in the third group, late i. p. chemotherapy (d 15, 20, 25 following surgery) was given via a port-a-cath aiming for significant reduction of a visible, already established peritoneal carcinomatosis. Mitomycin and cisplatin were highly effective to prevent peritoneal carcinomatosis (direct application immediately after tumor cell transfer - 1 (st) treatment group). Using early postoperative i. p. chemotherapy (2 (nd) group), 5-FU and CPT-11 were shown to be significantly effective to reduce the intraperitoneal tumor spread. None of the cytostatic agents was able to decrease significantly an already generated peritoneal carcinomatosis (3 (rd) treatment group). The results suggest that novel chemotherapeutic drugs should be proven for their potential to alter peritoneal metastases of GI tumors i) in comparison with established drugs and ii) depending on the application time and mode. PMID- 15354260 TI - Molecular mechanisms of megakaryocyte differentiation. AB - Each bone marrow megakaryocyte (MK) releases thousands of platelets into the circulation, and the underlying molecular and cellular mechanisms recently have received intense scrutiny. Genetic studies are beginning to clarify the mechanisms by which transcription factors help distinguish MK progenitors from other blood cell lineages and subsequently confer unique cellular properties. Other investigations demonstrate that platelets are assembled de novo during a terminal phase of MK differentiation in which the cell extends cytoplasmic projections known as proplatelets. This review focuses on the roles of selected transcription factors with key roles in MK differentiation, and on human and murine models of thrombocytopenia that result from impaired MK differentiation. The findings we review help construct a framework to appreciate thrombopoietic mechanisms in the context of underlying lineage and morphologic transitions. Many of these mechanisms are unique to MKs but appear to rely both on genes that are expressed only in that lineage and others that are expressed widely. PMID- 15354261 TI - Boundary events: contact-dependent and contact-facilitated signaling between platelets. AB - The theme of this review is that formation of a stable hemostatic plug requires adhesive interactions and signaling events that continue beyond the initial phases of platelet aggregation. These interactions and events are facilitated and, in some cases made possible, by the persistent close contacts between platelets that can only occur after the onset of aggregation. The molecules that are involved include integrins, cell adhesion molecules, receptor tyrosine kinases, and ligands that are either attached to or shed from the surface of activated platelets. The picture that emerges is one in which events after aggregation are nearly as complex as those that precede aggregation and the initiation of platelet plug formation. PMID- 15354262 TI - Platelet receptors for adenine nucleotides and thromboxane A2. AB - Adenosine diphosphate (ADP) and thromboxane A (2) (TXA (2)) are important physiological activators of platelets and exert their effects by acting on cell surface receptors. Platelet nucleotide receptors can be distinguished as three separate subtypes of the P2 receptor family. The P2X (1) receptor is a ligand gated adenosine triphosphate (ATP) receptor that was originally mistaken for an ADP receptor. This calcium-influx-causing receptor mediates platelet shape change and plays an important role in thrombus formation in small arterioles. The P2Y (1) receptor, through activation of G (q) and phospholipase C, is required for ADP-induced platelet shape change, fibrinogen receptor activation, and TXA (2) generation. The G (i)-coupled P2Y (12) receptor plays an important role in platelet aggregation, potentiation of dense granule release, and TXA (2) generation. Both the P2Y receptors are crucial for in vivo thrombus formation. TXA (2) stimulates two subtypes of G protein-coupled TP receptor, TPalpha and TPbeta, but its effects in platelets are mediated predominantly through the alpha isoform. Although interference with the activation of G protein-coupled ADP or TP receptors results in increased bleeding times and protection from thromboembolism, TP receptor antagonists did not translate into effective antiplatelet drugs. Blockade of ADP receptor is a mode of newer classes of antithrombotic drugs in the coming era. This review focuses on the contribution of different nucleotide receptors and TP receptors to platelet function and their potential as antithrombotic agents. PMID- 15354263 TI - Platelet-collagen responses: molecular basis and therapeutic promise. AB - Exposure of circulating blood to collagen after vessel injury has been recognized as a primary event in arterial thrombosis for many years but an understanding of the molecular basis of this response has only recently materialized. Platelet collagen interactions are initiated indirectly by interaction of platelet glycoprotein Ib (GPIb) with collagen-bound von Willebrand Factor (vWF). Slowed, rolling platelets firmly adhere following the generation of platelet activating signals in a process initiated by the platelet collagen receptor glycoprotein VI (GPVI). The contribution of the first platelet collagen receptor identified, integrin alpha (2)beta (1), remains controversial. alpha (2)beta (1) is not required for platelet responses to collagen under many experimental conditions but GPVI and other intracellular signals may activate the integrin, which is likely to play a later, secondary role. In vivo studies of arterial thrombosis using genetically modified mice suggest that blockade of platelet-collagen interactions may be a highly effective means of inhibiting arterial thrombosis. Recent studies have generated a more complete model of the molecular basis of platelet-collagen responses and provided both the means and incentive to create novel therapeutic agents aimed at blocking this process to treat human atherothrombotic diseases such as myocardial infarction and stroke. PMID- 15354264 TI - Protein-protein interactions in platelet alphaIIbbeta3 signaling. AB - The major platelet integrin alpha (IIb)beta (3) is the main receptor involved in platelet functions such as aggregation and spreading on extracellular matrix. Like all other integrins, alpha (IIb)beta (3) is capable of transducing signals both from inside and outside of the cell. To mediate these functions, alpha (IIb)beta (3) interacts with intracellular and transmembrane proteins. The identification of these proteins, as well as the study of their functions, has provided valuable insights into integrin-mediated function and signaling. This review summarizes the known proteins that directly interact with alpha (IIb)bbeta (3) and provides an overview of their roles in integrin function. PMID- 15354265 TI - Platelet secretory mechanisms. AB - Platelet granule secretion or exocytosis is required for normal platelet function and plays an important role in the pathogenesis of cardiovascular diseases. Platelets secrete molecules that amplify thrombosis, induce vascular remodeling, recruit and activate cells. The platelet secretory process begins in megakaryocytes where molecules are targeted to developing granules through specific vesicle trafficking and transporter mechanisms. Secretory granules may continue to mature in the circulation after the platelet has been released from the megakaryocyte. The platelet secretory process culminates when ligands interact with specific platelet receptors to trigger exocytosis. A convergence of new insights from several different organisms has begun to illuminate the molecular mechanisms responsible for the platelet secretory process, from granule development through membrane fusion and exocytosis. PMID- 15354266 TI - Platelet cyclic adenosine monophosphate phosphodiesterases: targets for regulating platelet-related thrombosis. AB - Platelets contain two cyclic adenosine monophosphate (cAMP) phosphodiesterases (PDEs) that regulate the level of cAMP, the major inhibitor of platelet activation pathways. PDE3A hydrolyzes cAMP to 5' AMP with a low K (m). PDE3A is inhibited by cyclic guanosine monophosphate (cGMP), which provides a feedback control and controls basal levels of cAMP. In contrast, PDE2A hydrolyzes both cAMP and cGMP with a high K (m), is allosterically stimulated by cGMP at moderate levels, and may control the stimulated levels of cAMP. Using affinity labeling, chemical modification, and site-directed mutagenesis of highly conserved amino acids, the amino acids required for catalytic activity and/or metal binding are H752 and H756. The singular binding sites for cAMP include N845, E971, and F972, whereas the unique amino acids interacting with cGMP are Y751, H836, H849, and D950. Residues E866 and F1004 are present in both the overlapping cGMP and cAMP sites. Two inhibitors of PDE3A are used in clinical medicine: milrinone and cilostazol. Three amino acids, Y751, D950, and F1004, show decreased sensitivity to both inhibitors (increased K (i)). These inhibitors mimic cGMP as an inhibitor of PDE3A rather than compete for cAMP binding. New nonhydrolyzable affinity labels inactivate PDE3A and are protected by Sp-cAMPS, a nonhydrolyzable substrate of the enzyme. These compounds have the potential to identify amino acids that are unique for PDE3A. An inhibitor of platelet PDE2A increases cAMP more than inhibitors of PDE3A but has much less effect on platelet activation, suggesting that these enzymes are present in different compartments of the cell. PMID- 15354267 TI - Platelet coagulation-protein interactions. AB - The biochemical mechanisms by which activated platelets participate in exposing receptors for the assembly of enzyme-cofactor-substrate complexes at all stages of the blood coagulation cascade are reviewed. Information derived from studies conducted during the last 30 years supports the concept that the initiation of blood coagulation is triggered by exposure of tissue factor at injury sites, leading to the generation of minute quantities of thrombin (limited by tissue factor pathway inhibitor), sufficient to activate platelets, factors XI, VIII, and V, and trigger the consolidation pathway (i.e., the sequential activation of factors XI, IX, X, and prothrombin on the activated platelet surface), leading to the generation of sufficient thrombin to convert fibrinogen to fibrin and effect hemostasis. Platelets localize coagulation to the hemostatic thrombus and protect coagulation enzymes from inhibition by both plasma and platelet inhibitors (e.g., protease nexin 2), thus preventing disseminated intravascular coagulation. PMID- 15354268 TI - Platelet transcriptome: the application of microarray analysis to platelets. AB - Human blood platelets are intimately involved in the regulation of thrombosis, inflammation, and wound repair. These cells retain megakaryocyte-derived cytoplasmic mRNA and functionally intact protein translational capabilities, although very little is known about normal or pathological mRNA profiles. Microarray analysis has demonstrated a clear and reproducible molecular signature unique to platelets. There is a relative paucity of expressed transcripts compared with those found in other eukaryotic cells, most likely related to mRNA decay in these anucleate cells. In contrast, a complementary methodology for transcript profiling (serial analysis of gene expression [SAGE]) demonstrates that 89% of tags represent mitochondrial (mt) transcripts (enriched in 16S and 12S ribosomal RNAs), presumably related to persistent mt-transcription in the absence of nuclear-derived transcripts. The abundance of nonmitochondrial SAGE tags parallels relative expression for the most abundant transcripts as determined by microarray analysis, establishing the concordance of both techniques for platelet profiling. These observations establish the validity of transcript analysis as a tool for identifying novel platelet genes that may regulate normal and pathologic platelet (and/or megakaryocyte) functions. The potential application of platelet-specific microarrays in scientific and clinical settings related to platelet production, cardiovascular, and cerebrovascular diseases is reviewed. PMID- 15354269 TI - Analyzing the platelet proteome. AB - During the last 10 years, mass spectrometry (MS) has become a key tool for protein analysis and has underpinned the emerging field of proteomics. Using high throughput tandem MS/MS following protein separation, it is potentially possible to analyze hundreds to thousands of proteins in a sample at a time. This technology can be used to analyze the protein content (i.e., the proteome) of any cell or tissue and complements the powerful field of genomics. The technology is particularly suitable for platelets because of the absence of a nucleus. Cellular proteins can be separated by either gel-based methods such as two-dimensional gel electrophoresis or one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by liquid chromatography (LC) -MS/MS or by multidimensional LC-MS/MS. Prefractionation techniques, such as subcellular fractionations or immunoprecipitations, can be used to improve the analysis. Each method has particular advantages and disadvantages. Proteomics can be used to compare the proteome of basal and diseased platelets, helping to reveal information on the molecular basis of the disease. PMID- 15354270 TI - Change in protein phenotype without a nucleus: translational control in platelets. AB - For most cells the nucleus takes center stage. Not only is it the largest organelle in eukaryotic cells, it carries most of the genome and transcription of DNA to RNA largely takes place in the nucleus. Because transcription is a major step in gene regulation, the absence of a nucleus is limiting from a biosynthetic standpoint. Consequently, the anucleate status of platelets has stereotyped it as a cell without synthetic potential. It is now clear, however, that this viewpoint is far too simplistic. In response to physiologic stimuli, platelets synthesize biologically relevant proteins that are regulated via gene expression programs at the translational level. This process does not require a nucleus; instead, it uses mRNAs and other translational factors that appear to be retained in specialized fashion as megakaryocytes generate platelets during thrombopoiesis. We highlight the molecular machinery and pathways used by platelets to translate mRNA into protein and offer insight into how these synthesized products may regulate thrombotic and inflammatory events. PMID- 15354272 TI - Drug interactions in gastroenterology: mechanisms, consequences, and how to avoid. AB - Drug interactions are an important avoidable cause of illness. With an increasing array of medications available to treat human disease and an increasing number of patients taking many medications, the risk of clinically significant drug interactions increases. This review describes some examples of common drug interactions in gastroenterology. The underlying mechanisms are discussed, and strategies are proposed to avoid drug interactions in clinical practice. PMID- 15354273 TI - Azathioprine, 6-mercaptopurine in inflammatory bowel disease: pharmacology, efficacy, and safety. AB - 6-Mercaptopurine and its prodrug azathioprine remain the mainstay of immunomodulator therapy for the maintenance of a steroid-free remission in patients with IBD. Recent evidence suggests that the cytotoxic and immunosuppressive effects of azathioprine might be mediated via the induction of lymphocyte apoptosis by its active metabolites, 6-thioguanine nucleotides. The therapeutic benefits of thiopurines have been shown to correlate with the concentration of 6-thioguanine nucleotides. Inherited differences in drug metabolism and disposition can significantly impact the safety and efficacy of these drugs. The thiopurine methyltransferase enzyme plays an important role in the metabolism of 6-mercaptopurine and azathioprine and in the determination of thiopurine cytotoxicity. By gaining an understanding of the pharmacology and metabolism of thiopurine therapy and putting it into the clinical context, clinicians will be able to optimize thiopurine therapy in IBD. PMID- 15354275 TI - Ex vivo histology-correlated optical coherence tomography in the detection of transmural inflammation in Crohn's disease. AB - BACKGROUND AND AIMS: Distinguishing Crohn's disease (CD) from ulcerative colitis (UC) can be difficult. Transmural inflammation, a key feature of CD, cannot be assessed by conventional colonoscopy with biopsy. Optical coherence tomography (OCT) provides high-resolution, cross-sectional images of the gut wall and might become a new diagnostic tool. The aims of this study were to perform histology correlated OCT on surgical specimens of CD and UC and to determine its diagnostic accuracy. METHODS: Colectomy specimens from patients with a preoperative diagnosis of CD (N = 24) or UC (N = 24) were studied with OCT in the operating room. OCT and histopathology were assessed blindly, and diagnostic accuracy of OCT was assessed. RESULTS: Eight preoperatively identified UC patients (33%) with transmural inflammation on postoperative histology were diagnosed with CD, and all 8 had a disrupted layered structure on OCT, a characteristic feature of transmural disease. Sixteen UC patients (67%) had superficial inflammation on histology; of them, 13 (81%) had an intact layered structure on OCT. All 24 preoperative CD patients had transmural inflammation on histology, and 23 (96%) had a disrupted layered structure on OCT. Of 585 histology-OCT image sets from the 48 patients, 152 sets (26%) had transmural inflammation on histology. The sensitivity and specificity for OCT to detect transmural disease were 86% and 91%, respectively. CONCLUSIONS: Transmural inflammation, as characterized by disruption of the layered structure of colon wall on OCT, is an accurate marker for the diagnosis of CD. Ex vivo OCT predicted transmural inflammation on postoperative histopathology. PMID- 15354274 TI - Optical biopsy: a new frontier in endoscopic detection and diagnosis. AB - Endoscopic diagnosis currently relies on the ability of the operator to visualize abnormal patterns in the image created by light reflected from the mucosal surface of the gastrointestinal tract. Advances in fiber optics, light sources, detectors, and molecular biology have led to the development of several novel methods for tissue evaluation in situ. The term "optical biopsy" refers to methods that use the properties of light to enable the operator to make an instant diagnosis at endoscopy, previously possible only by using histological or cytological analysis. Promising imaging techniques include fluorescence endoscopy, optical coherence tomography, confocal microendoscopy, and molecular imaging. Point detection schemes under development include light scattering and Raman spectroscopy. Such advanced diagnostic methods go beyond standard endoscopic techniques by offering improved image resolution, contrast, and tissue penetration and providing biochemical and molecular information about mucosal disease. This review describes the basic biophysics of light-tissue interactions, assesses the strengths and weaknesses of each method, and examines clinical and preclinical evidence for each approach. PMID- 15354276 TI - Is there a relationship between obstructive sleep apnea and gastroesophageal reflux disease? AB - BACKGROUND AND AIMS: The causative relationship between obstructive sleep apnea (OSA) and gastroesophageal reflux disease (GERD) remains an area of controversy. The aim of the study is to further explore the relationship between OSA and GERD in a large group of patients with OSA. METHODS: One hundred thirty-six patients referred for polysomnographic studies at the University of Arizona Sleep Center were included in this study. All patients were assessed by means of a demographic survey, the validated GERD Symptom Checklist, and the Sleep Heart Health Study Sleep Habits Questionnaire. Polysomnograms were scored for objective measures of sleep and breathing. OSA is defined as an apnea-hypopnea index > 5 with compatible symptoms. RESULTS: Of the 136 subjects, 101 (74%) were found to have OSA. Self-report of heartburn or acid regurgitation symptoms was unrelated to severity of sleep apnea. Additionally, OSA was not influenced by the severity of GERD. Objective measures of disordered sleep had stronger associations with age, smoking, and alcohol use than GERD in men and stronger associations with age and body mass index than GERD in women. Subjectively reported sleep quality was affected more strongly by GERD severity than by age, smoking, alcohol use, or the presence of OSA. Only females administered antireflux medications were less likely to report poor sleep quality. CONCLUSIONS: Subjective reports of sleep quality were affected by GERD severity, but an objective correlation between OSA and GERD was lacking. This may suggest that GERD and OSA are common entities that share similar risk factors, but appear not to be causally linked. PMID- 15354277 TI - Responsiveness and interpretation of a symptom severity index specific to upper gastrointestinal disorders. AB - BACKGROUND AND AIMS: Determining clinically meaningful change of patient-reported outcome measures is important for evaluating effectiveness of treatments for gastrointestinal (GI) diseases. This study evaluates responsiveness of the Patient Assessment of Gastrointestinal Disorders-Symptom Severity Index (PAGI SYM) in gastroesophageal reflux disease (GERD) and dyspepsia. METHODS: The PAGI SYM was based on a review of the published literature and interviews with patients and clinicians. Items were developed to be linguistically and culturally appropriate for multicountry studies. The PAGI-SYM includes 6 subscales: heartburn/regurgitation, fullness/early satiety, nausea/vomiting, bloating, upper abdominal pain, and lower abdominal pain. Subjects with GERD (n = 810) or dyspepsia (n = 767) participated in this multicountry, observational study. All subjects completed the PAGI-SYM, a global symptom relief questionnaire, and a measure of patient-rated change in GI-related symptoms, the Overall Treatment Effect (OTE) scale. Responsiveness was evaluated at 8 weeks by comparing groups by disease, symptom relief, and OTE (improved, stable, and worsened). RESULTS: Subjects reporting symptom relief reported significantly lower (better) PAGI-SYM scores than those reporting no symptom relief ( P < 0.0001 to P < 0.0005). Subjects with improvements in overall GI symptoms exhibited significant decreases in PAGI-SYM subscale scores compared with those who remained the same or worsened (all P values < 0.0001). Effect sizes ranged from 0.21-1.28, and standard errors of measurement ranged from 0.29-0.63, depending on subscale and disease sample. CONCLUSIONS: The PAGI-SYM is a brief symptom severity instrument that measures common GI symptoms. Results suggest that the PAGI-SYM is responsive and sensitive to change in clinical status in subjects with GERD or dyspepsia. PMID- 15354278 TI - Responsiveness and interpretation of a quality of life questionnaire specific to upper gastrointestinal disorders. AB - BACKGROUND AND AIMS: The Patient Assessment of Upper Gastrointestinal Disorders Quality of Life (PAGI-QOL) has been developed and validated to assess quality of life in gastroesophageal reflux disease, dyspepsia, and gastroparesis. The aim of this work was to assess responsiveness of the PAGI-QOL to clinical changes and provide guidance for the interpretation of score changes. METHODS: The analysis was based on 2 observational multicenter, longitudinal, 8-week observation studies, one in the United States and one in 5 European countries. The Overall Treatment Effect scale completed by patients and clinicians at 8 weeks was used to define groups of improved, worsened, and stable patients. Responsiveness was assessed by describing the change in PAGI-QOL scores for each of these groups, comparing these changes over time, and calculating effect sizes and standardized response means. These data were used to define the minimal clinically important difference. RESULTS: A total of 1736 patients with upper gastrointestinal disorders were analyzed. High sensitivity to change over time was demonstrated, with effect sizes and standardized response means in improved patients for the PAGI-QOL total score ranging from 0.79-1.41. A change of 0.4 for the PAGI-QOL total score might be considered as the minimal clinically important difference when comparing pretreatment and post-treatment PAGI-QOL total scores. CONCLUSIONS: The PAGI-QOL questionnaire is a responsive and clinically relevant instrument for assessing quality of life in patients with upper gastrointestinal disorders. PMID- 15354279 TI - The presentation of irritable bowel syndrome in the context of somatization disorder. AB - BACKGROUND AND AIMS: Somatization disorder, a female predominant disorder, has been found with higher than expected prevalence in previous studies of irritable bowel syndrome (IBS) and might be responsible for some of the comorbidity and excessive health care resource use associated with IBS. The study's aim was to determine within a female IBS population the degree of segregation of psychiatric illness, functional disorders, and health care utilization with somatization disorder. METHODS: In a prospective, 6-month follow-up study, psychiatric disorders were assessed with the Diagnostic Interview Schedule, gastrointestinal and other symptoms with self-report questionnaires, and medically unexplained complaints by thorough chart review. The setting was a university gastroenterology clinic. The participants were a convenience sample of female clinic attendees with IBS (N = 56). RESULTS: Somatization disorder was diagnosed in 25% of patients and highly probable in another 5%. Somatization disorder was associated with significantly greater numbers of gastrointestinal and other symptoms, psychiatric disorders, physicians consulted, telephone calls to physicians, urgent care visits, medication changes, and missed work days and with benzodiazepine use. On follow-up, somatization disorder was associated with psychiatric and IBS symptoms, medication changes, and treatment dissatisfaction. Both somatization disorder and other psychiatric illnesses were associated with other functional gastrointestinal disorders; only somatization disorder remained predictive in a regression model that controlled for the presence of other psychiatric illness. CONCLUSIONS: Among female IBS patients attending a university gastroenterology clinic, many aspects of comorbidity and health care behaviors previously associated with IBS segregated with the diagnosis of somatization disorder. Recognition and appreciation of somatization disorder in IBS have important ramifications for the conduct of research and clinical practice. PMID- 15354280 TI - Effect of tegaserod in chronic constipation: a randomized, double-blind, controlled trial. AB - BACKGROUND AND AIMS: Chronic constipation is a common gastrointestinal disorder. The aim of this study was to evaluate the efficacy, safety, and tolerability of tegaserod, a serotonin subtype 4 receptor partial agonist in patients with chronic constipation. METHODS: This was a randomized, double-blind, placebo controlled study. After a 2-week baseline, patients received tegaserod 2 mg twice daily (n = 450), tegaserod 6 mg twice daily (n = 451), or placebo (n = 447) for 12 weeks, followed by a 4-week withdrawal period. Responders were those patients having been treated for at least 7 days with an increase of > or =1 complete spontaneous bowel movement/week vs. baseline during weeks 1-4 (primary variable) and weeks 1-12 (secondary variable). Other secondary variables included patient assessment of constipation symptoms (number of bowel movements, stool form, abdominal bloating/distention, straining, and abdominal pain/discomfort), and global assessment of constipation and bowel habits. RESULTS: Responder rates for complete spontaneous bowel movement during weeks 1-4 were significantly greater ( P < 0.0001) in the tegaserod 2 mg twice daily (41.4%) and 6 mg twice daily groups (43.2%) vs. placebo (25.1%). This effect was maintained over 12 weeks. Statistically significant improvements over placebo were observed across the majority of secondary variables for both tegaserod doses. No rebound effect was observed after treatment withdrawal. Tegaserod was well tolerated; headache and nasopharyngitis, the most frequent adverse events, were more common in the placebo group than in either tegaserod group. CONCLUSIONS: Over 12 weeks, tegaserod treatment produced significant improvements in chronic constipation symptoms and was also safe and well tolerated. PMID- 15354281 TI - A genetic contribution to inflammatory bowel disease in Iceland: a genealogic approach. AB - BACKGROUND AND AIMS: Both genetic and environmental factors play a role in the development of Crohn's disease (CD) and ulcerative colitis (UC), collectively known as inflammatory bowel disease (IBD). The aim of this study was to estimate the genetic component in IBD in Iceland. METHODS: A population-based sample, representing everyone diagnosed with IBD in Iceland from 1950 to 1996, was studied using a computerized population-wide genealogic database. The relationships among the patients were analyzed by calculating the kinship coefficient and the relative risk. RESULTS: The kinship coefficients for the patients were significantly greater than the mean kinship coefficient for the controls ( P < 10 -6 ). The risk ratio for siblings of IBD, UC, and CD patients was 5.0 ( P < 0.001), 5.9 ( P < 0.001), and 4.1 ( P = 0.033), respectively. The cross-risk ratio for siblings of UC patients developing CD (or vice versa) was 2.6 ( P = 0.015). CONCLUSIONS: The results indicate that the IBD patients are more closely related than the controls, which strongly supports the involvement of a genetic component in the development of IBD in Icelandic patients. We find that the increase in risk for relatives of UC probands to develop UC, or relatives of CD probands to develop CD, is greater than the increase in risk for relatives of UC probands to develop CD, or relatives of CD probands to develop UC. PMID- 15354282 TI - Physician assessment of family cancer history and referral for genetic evaluation in colorectal cancer patients. AB - BACKGROUND AND AIMS: An accurate family history is an essential component of cancer risk assessment. Our aim was to determine the concordance of family history assessments made by physicians with patients' self-reports and the frequency of referral for genetic evaluation in high-risk colorectal cancer (CRC) patients. METHODS: A self-administered family cancer history questionnaire was completed by 387 consecutive CRC patients at their first visit to a gastroenterology cancer clinic. Physician notes from the first visit were reviewed to determine the concordance of the family cancer history with patients' self-reported history. Prevalence of individuals that satisfied the Bethesda guidelines for hereditary colon cancer were compared with actual rates of referral. Regression analyses were used to determine factors associated with a comprehensive physician evaluation of family history. RESULTS: Oncologists documented a comprehensive family history in 59% (184 of 311) of patients with a first- or second-degree relative with cancer. Young age at diagnosis and a first degree relative with CRC were not associated with a more comprehensive family history assessment. An increasing number of cancers per family was a strong predictor of a less comprehensive family history assessment (odds ratio = 0.63; P < 0.0001). Seventy-five of 387 (19%) CRC patients met Bethesda guidelines for genetics assessment, however, only 13 of 75 (17%) were referred. CONCLUSIONS: Increased complexity in family cancer history leads to a decrease in accuracy of family history, suggesting the need for systematic approaches to facilitate family history assessment. Familial cancer risk remains largely unrecognized and referral rates for genetic evaluation for CRC syndromes are low. PMID- 15354283 TI - Role of ethnicity in risk for hepatocellular carcinoma in patients with chronic hepatitis C and cirrhosis. AB - BACKGROUND AND AIMS: In the United States, hepatocellular carcinoma (HCC) is more common among Asians and African Americans than Caucasians, with chronic hepatitis C virus (HCV) infection accounting for up to half of the patients. Our study examined ethnicity as a potential risk factor for HCC among patients with chronic hepatitis C. METHODS: We conducted a case-control study of 464 patients with chronic hepatitis C and cirrhosis (207 cancer patients and 257 controls) using medical records and pathology records at 4 medical centers. We estimated odds ratios with 95% confidence intervals by using conditional logistic regression on case-control sets, matched within study centers and study period on sex and age groups (< or =45, 46-55, 56-65, >65 yr). To control for potential confounding caused by severity of cirrhosis and residual confounding caused by age, we also included Child-Turcotte-Pugh (CTP) scores and age (continuous variable) in all regression analyses. RESULTS: Compared with Caucasians, the cancer risk was increased significantly among Asians (adjusted odds ratio, 4.3; 95% confidence interval, 2.1-9.0 for men, and 4.6; 1.2-18.5 for women) and somewhat increased among African-American men (adjusted odds ratio, 2.4; 95% confidence interval, 0.9-6.3). CONCLUSIONS: This study suggests that, among patients with chronic hepatitis C and cirrhosis, liver cancer risk is increased 4-fold in Asians and may be doubled in African-American men, compared with Caucasians. These results need confirmation in larger studies from racially diverse populations, but, if confirmed, these results point to high-risk populations that should be targeted for screening and preventive efforts. PMID- 15354284 TI - Features and outcome of autoimmune hepatitis type 2 presenting with isolated positivity for anti-liver cytosol antibody. AB - BACKGROUND AND AIMS: Autoimmune hepatitis (AIH) type 2 is identified by the presence in the serum of anti-liver/kidney microsome type 1 autoantibody. Anti liver cytosol autoantibody has been reported in children with autoimmune liver disorders mostly in association with anti-liver/kidney microsome reactivity. However, its role as a sole marker of AIH type 2 is debated. We describe here a series of 18 children and adolescents (15 girls, 3 boys) with AIH with serum anti liver cytosol type 1 (aLC1) as the only autoimmune marker. METHODS: A retrospective review was conducted from 3 pediatric hepatology units of all children with an autoimmune liver disease associated with aLC1 as found by immunofluorescence and/or immunodiffusion or immunoblotting. RESULTS: Age at first symptoms ranged from 11 months to 14 years; 12 children presented with acute hepatitis, 1 with progressive jaundice, and 5 were asymptomatic. Anti liver/kidney microsome, antimitochondria, and anti-actin autoantibodies were not detected. Signs of cirrhosis were present in 11 children. Immunosuppressive treatment was effective in all except 2 children who had subfulminant hepatic failure and who required liver transplantation. Sixteen patients (14 with their native liver) currently are alive; 14 patients still are on immunosuppressive therapy after 1 to 22 years. According to the international scoring system for the diagnosis of AIH, 16 patients corresponded to a definite diagnosis and 2 corresponded to a probable diagnosis. CONCLUSIONS: The presence of aLC1 in children with acute or chronic liver disease of unknown origin strongly supports a diagnosis of AIH and is an indication for early immunosuppressive therapy. PMID- 15354285 TI - Trends in incidence of acute pancreatitis in a Swedish population: is there really an increase? AB - BACKGROUND AND AIMS: Recent reports have suggested an increasing incidence of acute pancreatitis, and changing patterns of risk factors, over the past decades. The aim of this study was to investigate trends in the incidence of acute pancreatitis, and risk factors related to the disease, in a general population over a 15-year period. METHODS: Clinical, autopsy, and forensic records for all patients with a first attack of acute pancreatitis in Malmo, Sweden, from 1985 to 1999, were validated retrospectively. Evidence for diagnosis was reconsidered and plausible cause was assessed. The incidence of gallstone disease, lung cancer, and alcohol-related conditions in the background population were retrieved from hospital diagnosis records and cancer and cause-of-death registries. RESULTS: A total of 929 first attacks of acute pancreatitis were identified. The total incidence of acute pancreatitis increased by 3.9% per year (95% confidence interval [CI], 2.1-5.8). The incidence of gallstone-related pancreatitis increased by 7.6% per year (95% CI, 4.0-11.4), and this correlated with an increase in the incidence of other gallstone-related conditions ( r = 0.68; P = 0.005). Alcohol-related pancreatitis decreased by -5.1% per year (95% CI, -7.4 to -2.8), and this correlated with a decrease in the incidence of delirium tremens ( r = 0.75; P = 0.001), mortality from cirrhosis ( r = 0.81; P < 0.001), and incidence of lung cancer ( r = 0.57; P = 0.026). CONCLUSIONS: There was a statistically significant increase in the incidence of acute pancreatitis. Gallstone-related pancreatitis increased, and alcohol-related pancreatitis decreased. Both of these trends were statistically significant and correlated with trends in the incidence of other conditions associated with either gallstone disease or alcohol abuse. PMID- 15354286 TI - The usefulness of PPD testing in inflammatory bowel disease patients before infliximab therapy. PMID- 15354287 TI - Cre-loxP mediated control of PrP to study transmissible spongiform encephalopathy diseases. AB - Expression of the PrP glycoprotein is essential for the development of the transmissible spongiform encephalopathy (TSE) or prion diseases. Although PrP is widely expressed in the mouse, the precise relevance of different PrP-expressing cell types to disease remains unclear. To address this, we generated two lines of floxed PrP gene-targeted transgenic mice using the Cre recombinase-loxP system. These floxed mice allow a functional PrP allele to be either switched "on" or "off." We demonstrate control of PrP expression for both alleles following Cre mediated recombination, as determined by PrP mRNA and protein expression in the brain. Moreover, we show that Cre-mediated alteration of PrP expression in these mice has a major influence on the development of TSE disease. These floxed PrP mice will allow the involvement of PrP expression in specific cell types following TSE infection to be defined, which may identify potential sites for therapeutic intervention. PMID- 15354288 TI - Generation of a conditional allele of the mouse prostaglandin EP4 receptor. AB - Genetic disruption of the mouse EP4 receptor results in perinatal lethality associated with persistent patent ductus areteriosus (PDA). To circumvent this, an EP4 allele amenable to conditional deletion using the Cre/loxP system was generated. The targeting construct was comprised of a floxed exon2 in tandem with the neomycin-resistance gene in intron 2, flanked by third 3' LoxP site. Mice homozygous for the targeted allele (EP4(lox+neo/lox+neo)), or following its Cre mediated deletion (EP4(del/del)), also die within hours of birth with PDA. In contrast, mice homozygous for a partially recombined allele, retaining exon2 but lacking neo (EP4(flox/flox)), are viable and show no overt phenotype. Postnatal deletion of the floxed EP4 gene is efficiently achieved in the liver and kidney in a transgenic mouse expressing the inducible Mx1Cre recombinase. The EP4(flox) mouse should provide a useful reagent with which to examine the physiologic roles of the EP4 receptor. PMID- 15354289 TI - Delayed degradation of parental macronuclear DNA in programmed nuclear death of Paramecium caudatum. AB - In the ciliated protozoan Paramecium caudatum, a parental macronucleus that is fragmented into some 40-50 pieces during conjugation does not degenerate immediately, but persists until the eighth cell cycle after conjugation. Here we demonstrate that the initiation of the parental macronuclear degeneration occurs at about the fifth cell cycle. The size of parental macronuclear fragments continued to increase between the first and fourth cell cycle, but gradually decreased thereafter. By contrast, a new macronucleus grew and reached a maximum size by the fourth cell cycle, suggesting that the new macronucleus matured by that stage. Southern blot analysis revealed that parental macronuclear DNA was degraded at about the fifth cell cycle. The degradation was supported by acridine orange staining, indicating degeneration of the macronuclear fragments. Prior to the degradation, the fragments once attached to the new macronucleus were subsequently liberated from it. These observations lead us to conclude that once a new macronucleus has been fully formed by the fourth cell cycle, the parental macronuclear fragments are destined to degenerate, probably through direction by new macronucleus. Considering the long persistence of the parental macronucleus during the early cell cycles after conjugation, the macronuclear fragments might function in the maturation of the imperfect new macronucleus. Two possible functions, a gene dosage compensation and adjustment of ploidy level, are discussed. PMID- 15354290 TI - Programmed cell death in Bradysia hygida (Diptera, Sciaridae) salivary glands presents apoptotic features. AB - In this work, we present biochemical and morphological evidence that the final steps of programmed cell death (PCD) in the salivary glands of the inferior Diptera, Bradysia hygida, present apoptotic characteristics. In B. hygida, elimination of salivary glands is preceded by the establishment of a typical pattern of protein synthesis; increase in caspase activity; decrease in cell volume; nuclear pyknosis; nuclear DNA breakage; changes in the actin cytoskeleton; and most importantly, destruction of giant cells via formation of apoptotic bodies containing broken DNA or cytoplasm remains. Thus, elimination of B. hygida salivary glands by this process suggests that such mode of PCD is also involved in the destruction of entire organs in insects and, therefore, adds more complexity to the regulation of tissue elimination during development. PMID- 15354291 TI - Frizzled-9 promoter drives expression of transgenes in the medial wall of the cortex and its chief derivative the hippocampus. AB - The hippocampus develops from the medial wall of the forming cerebral cortex during embryonic life. Morphogenic signals from the Wnt pathway regulate several events during hippocampal development (Galceran et al.: Development 127:469-482, 2000; Lee et al.: Development 127:457-467, 2000; Zhou et al.: J Neurosci 24:121 126, 2004) and we have previously shown that Wnt receptors from the Frizzled (Fzd) family are expressed in discreet cortical domains during development (Kim et al.: Mech Dev 103:167-172, 2001). We generated transgenic mice using the putative control elements of the Fzd9 gene, normally selectively expressed in the developing and adult hippocampus, driving expression of a marker gene. These mice express LacZ in the brain in the same developmental distribution as endogenous Fzd protein. Postnatally, expression remains strong in the dendritic fields of hippocampal principal cells as well as hippocampal efferent axons. These mice provide a genetic and anatomic tool for analyzing development and reorganization in the hippocampus. PMID- 15354292 TI - Notch pathway is dispensable for adipocyte specification. AB - In the past decade we have witnessed an epidemic of obesity in developed countries. Therefore, understanding the mechanisms involved in regulation of body weight is becoming an increasingly important goal shared by the public and the scientific community. The key to fat deposition is the adipocyte, a specialized cell that plays a critical role in energy balance and appetite regulation. Much of our knowledge of adipogenesis comes from studies using preadipocytic cell lines that have provided important information regarding molecular control of adipocyte differentiation. However, they fall short of revealing how naive cells acquire competence for adipogenesis. Studies in preadipocytes indicate that the Notch pathway plays a role in regulating adipogenesis (Garces et al.: J Biol Chem 272:29729-29734, 1997). Given the known biological functions of Notch in mediating cell fate decisions (Artavanis-Tsakonas et al.: Science 284:770-776, 1999), we wished to test the hypothesis that the Notch pathway is required for this cellular program by examining adipogenesis in several genetic loss-of function models that encompass the entire pathway. We conclude that the "canonical" Notch signaling pathway is dispensable for adipocyte specification and differentiation from either mesenchymal or epithelial progenitors. PMID- 15354293 TI - Highly restricted expression of Cre recombinase in cerebellar Purkinje cells. AB - The Purkinje neuron, one of the most fascinating components of the cerebellar cortex, is involved in motor learning, motor coordination, and cognitive function. Purkinje cell protein 2 (Pcp2/L7) expression is highly restricted to Purkinje and retinal bipolar cells, where it has been exploited to enable highly specific, Cre recombinase-mediated, site-specific recombination. Previous studies showed that mice carrying a Cre transgene produced by insertion of Cre cDNA into a small 2.88-kb Pcp2 DNA fragment expressed Cre in Purkinje cells; however, some Cre activity was also observed outside the target tissues. Here, we used Red mediated recombineering to insert Cre cDNA into a 173-kb BAC carrying the entire intact Pcp2 gene, and characterize the resultant BAC/Cre transgenic mice for Cre expression. We show that BAC/Cre transgenic mice have exclusive Cre expression in Purkinje and bipolar cells and nowhere else. These mice will facilitate Purkinje cell and retinal bipolar cell-specific genetic manipulation. PMID- 15354294 TI - Generation of a conditional knockout allele for the Janus kinase 2 (Jak2) gene in mice. AB - To study biologically relevant functions of the Janus kinase 2 (Jak2) in multiple cytokine and hormone receptor signal transduction pathways, we generated a conditional knockout (floxed) allele of this gene by placing loxP sites around the first coding exon of Jak2. Homozygous floxed animals developed normally and exhibited no phenotypic abnormalities. The conversion of the floxed allele into a null mutation was achieved by transmitting the targeted allele through the female germline of MMTV-Cre (line A) mice. Embryos that carry two Jak2 null alleles died around midgestation and exhibited impaired definitive erythropoiesis, which is a hallmark of Jak2 deficiency reported previously in conventional knockouts. This observation suggested that the Cre-mediated deletion of the first coding exon results in a true null mutation that is incapable of mediating signals through the erythropoietin receptor. Using mouse embryonic fibroblasts derived from Jak2 null embryos and their wildtype littermate controls, we demonstrated that Jak2 deficiency decouples growth hormone-receptor signaling from its downstream mediators, the signal transducer and activator of transcription (Stat) 5a and 5b. PMID- 15354296 TI - Multiple passages of light through an absorption inhomogeneity in optical imaging of turbid media. AB - Multiple passages of light through an absorption inhomogeneity of finite size deep within a turbid medium are analyzed for optical imaging by use of the self energy diagram. The nonlinear correction becomes more important for an inhomogeneity of a larger size and with greater contrast in absorption with respect to the host background. The nonlinear correction factor agrees well with that from Monte Carlo simulations for cw light. The correction is approximately 50%-75% in the near infrared for an absorption inhomogeneity with the typical optical properties found in tissues and five times the size of the transport mean free path. PMID- 15354295 TI - RNA polymerase II 140wimp mutant and mutations in the TFIIH subunit XPB differentially affect homeotic gene expression in Drosophila. AB - Mutations in the XPB and XPD helicases of the DNA repair/transcription factor TFIIH are involved in several human genetic disorders. An unanswered problem concerning the complexity of the phenotype-genotype relationship is why mutations in individual subunits of TFIIH produce specific phenotypes and not many others. In order to investigate this question we tested whether mutations in the Drosophila XPB homolog, haywire (hay), would modify homeotic derepression phenotypes. In this work, we report that mutations in hay and in the 140-kDa subunit of the RNA polymerase II (RpII140wimp) act as dominant modifiers of the derepression phenotypes of the Sex combs reduced (Scr) and Ultrabithorax (Ubx) genes. The hay mutations only weakly suppress the Scr derepression phenotype caused by the Antp(Scx) mutation but not by Polycomb. In contrast, the RpII140wimp mutation strongly suppresses both Scr derepression phenotypes. In addition, the RpII140wimp also generates phenotypes indicative of loss of Ubx function. On the other hand, all the derepression homeotic phenotypes are sensitive to the generalized reduction of transcription levels when the flies are grown with actinomycin D. We also show that different promoter control regions have differential sensitivity to different hay alleles. All these results support that although TFIIH is a basal transcription factor, mutations in the subunit encoded by hay have specific effects in the transcription of some genes. PMID- 15354297 TI - [Public health problem of psychological stress and its preventive measures]. PMID- 15354298 TI - [Environmental hormones and their effects on human health]. PMID- 15354299 TI - [Progress on the study of DNA-dependent protein kinase complex]. PMID- 15354300 TI - Evolutionary and biogeographic patterns of the Badidae (Teleostei: Perciformes) inferred from mitochondrial and nuclear DNA sequence data. AB - We reconstructed phylogenetic relationships of the family Badidae using both mitochondrial and nuclear nucleotide sequence data to address badid systematics and to evaluate the role of vicariant speciation on their evolution and current distribution. Phy-logenetic hypotheses were derived from complete cytochrome b (1,140 base pairs) sequences of 33 individuals representing 13 badid species, and using three species of Nandidae as outgroups. Additionally, we sequenced the nuclear RAG1 (1,473 base pairs) and Tmo-4C4 (511 base pairs) genes from each of the badid species and one representative of the outgroup. Our molecular data provide the first phylogenetic hypothesis of badid intrarelationships. Analysis of the mitochondrial and nuclear nucleotide sequence data sets resulted in well supported trees, indicating a basal split between the genera Dario and Badis, and further supporting the division of the genus Badis into five species groups as suggested by a previous taxonomic revision of the Badidae. Within the genus Badis, mitochondrial and nuclear phylogenies differed in the relative position of B. kyar. We also used our molecular phylogeny to test a vicariant speciation hypothesis derived from geological evidence of large-scale changes in drainage patterns in the Miocene affecting the Irrawaddy- and Tsangpo-Brahmaputra drainages, in the southeastern Himalaya. Within both genera, Badis and Dario, we observed a divergence into Irrawaddy- and Tsangpo-Brahmaputra clades. Using a cytb substitution rate of 8.2 x 10(-9) (substitutions x base pair(-1) x year(-1), we tentatively date this vicariant event at the Oligocene-Miocene boundary (19 24Myr). It is concordant with a hypothesized paleo connection of the Tsangpo river with the Irrawaddy drainage that was most likely interrupted during Miocene orogenic events through tectonic uplifts in eastern Tibet. Our data, therefore, indicate a substantial role of vicariant-based speciation shaping the current distribution patterns of badids. PMID- 15354301 TI - Effects of incorrect computer-aided detection (CAD) output on human decision making in mammography. AB - RATIONALE AND OBJECTIVES: To investigate the effects of incorrect computer output on the reliability of the decisions of human users. This work followed an independent UK clinical trial that evaluated the impact of computer-aided detection(CAD) in breast screening. The aim was to use data from this trial to feed into probabilistic models (similar to those used in "reliability engineering") which would detect and assess possible ways of improving the human CAD interaction. Some analyses required extra data; therefore, two supplementary studies were conducted. Study 1 was designed to elucidate the effects of computer failure on human performance. Study 2 was conducted to clarify unexpected findings from Study 1. MATERIALS AND METHODS: In Study 1, 20 film readers viewed 60 sets of mammograms (30 of which contained cancer) and provided "recall/no recall" decisions for each case. Computer output for each case was available to the participants. The test set was designed to contain an unusually large proportion (50%) of cancers for which CAD had generated incorrect output. In Study 2, 19 different readers viewed the same set of cases in similar conditions except that computer output was not available. RESULTS: The average sensitivity of readers in Study 1 (with CAD) was significantly lower than the average sensitivity of read-ers in Study 2 (without CAD). The difference was most marked for cancers for which CAD failed to provide correct prompting. CONCLUSION: Possible automation bias effects in CAD use deserve further study because they may degrade human decision-making for some categories of cases under certain conditions. This possibility should be taken into account in the assessment and design of CAD tools. PMID- 15354302 TI - Automatic path searching for interactive navigation support within virtual medical 3-dimensional objects. AB - RATIONALE AND OBJECTIVES: This article proposes the use of a disembodied autonomous actor for navigation support within complex virtual medical objects reconstructed from Computed Tomography or Magnetic Resonance Imaging. Such objects are often maze-like, and users risk getting lost within them during Virtual Reality sessions. Therefore, users need paths for guided fly-throughs when performing non-invasive diagnostic tasks. MATERIALS AND METHODS: We present a synthetic vision-based actor capable of finding collision-free paths from a given position to a goal point in environments containing loops and impasses. When navigating, the actor voxelizes the virtual environment and searches for collision-free paths in voxel space by using a back tracking search algorithm. Automata and rules control its search behaviour. The resulting paths can be used in dedicated virtual endoscopy applications. RESULTS: Our path search method has been tested within a variety of tubular virtual anatomical structures in 3D such as aortas, colons, or blood vessels of the brain. The actor finds paths within reasonable time limits, even when considering complex anatomical surface models. CONCLUSION: The method may be used as a valuable tool for assisting virtual endoscopic diagnostic and screening activities in the near future. PMID- 15354303 TI - "B" readers' radiographic interpretations in asbestos litigation: is something rotten in the courtroom? PMID- 15354304 TI - Comparison of "B" readers' interpretations of chest radiographs for asbestos related changes. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine if chest radiographic interpretations by physicians retained by attorneys representing persons alleging respiratory changes from occupational exposure to asbestos would be confirmed by independent consultant readers. MATERIALS AND METHODS: For 551 chest radiographs read as positive for lung changes by initial "B" readers retained by plaintiffs' attorneys, 492 matching interpretative reports were made available to the authors. Six consultants in chest radiology, also B readers, agreed to re-interpret the radiographs independently without knowledge of their provenance. The film source, patient name, and other identifiers on each film were masked. The International Labor Office 1980 Classification of Chest Radiographs(ILO 80) was used with forms designed by the US National Institute of Occupational Safety and Health to record the consult-ants' findings. The results were compared with initial readings for film quality, complete negativity, parenchymal abnormalities,small opacities profusion, and pleural abnormalities using chi-square tests and kappa statistics.Results. Initial readers interpreted study radiographs as positive for parenchymal abnormalities (ILO small opacity profusion category of 1/0 or higher) in 95.9% of 492 cases. Six consultants classified the films as 1/0 or higher in 4.5% of 2,952 readings. Statistical tests of these and other comparable data from the study showed highly significant differences between the interpretations of the initial readers and the findings of the consultants. CONCLUSION: The magnitude of the differences between the interpretations by initial readers and the six consultants is too great to be attributed to interobserver variability. There is no support in the literature on x-ray studies of workers exposed to asbestos and other mineral dusts for the high level of positive findings recorded by the initial readers in this report. PMID- 15354305 TI - MR imaging of the prostate at 3 Tesla: comparison of an external phased-array coil to imaging with an endorectal coil at 1.5 Tesla. AB - RATIONALE AND OBJECTIVES: To qualitatively compare the image quality of torso phased-array 3-Tesla (3T) imaging of the prostate with that of endorectal 1.5 Tesla imaging. MATERIALS AND METHODS: Twenty cases of torso phased-array prostate imaging performed at 3-Tesla with FSE T2 weighted images were evaluated by two readers independently for visualization of the posterior border (PB), seminal vesicles (SV), neurovascular bundles (NVB), and image quality rating (IQR). Studies were performed at large fields of view(FOV) (25 cm) (14 cases) (3TL) and smaller FOV (14 cm) (19 cases) (3TS). A comparison was made to 20 consecutive cases of 1.5-T endorectal evaluation performed during the same time period.Results. 3TL produced a significantly better image quality compared with the small FOV for PB (P = .0001), SV (P =.0001), and IQR (P = .0001). There was a marginally significant difference within the NVB category (P = .0535). 3TL produced an image of similar quality to image quality at 1.5 T for PB (P = .3893), SV (P = .8680), NB (P = .2684), and IQR (P = .8599). CONCLUSION: Prostate image quality at 3T with a torso phased-array coil can be comparable with that of endorectal 1.5-T imaging. These findings suggest that additional options are now available for magnetic resonance imaging of the prostate gland. PMID- 15354306 TI - Practical ethics. Self-promotion. PMID- 15354307 TI - Mutations correlated to NVP concentrations. PMID- 15354308 TI - Resistance profiles of new antiretrovirals. PMID- 15354309 TI - Dialysis machine alert. PMID- 15354310 TI - Vaccine trial underway in Thailand. PMID- 15354311 TI - Preimplantation HLA typing and stem cell transplantation: report of International Meeting, Cyprus, 27-8 March, 2004. AB - There has been progress in the application of stem cell transplantation for the treatment of an increasing number of severe congenital and acquired bone marrow disorders that are currently restricted by the availability of human leukocyte antigen(HLA)-matched related donors. Preimplantation HLA typing has recently been introduced to improve the access to stem cell therapy for inherited bone marrow failures, and its possible use for the treatment of common sporadic malignant and non-malignant bone marrow disorders has also been explored. This paper describes the current experience of preimplantation HLA typing, reviewed by the International Meeting on the subject, which includes preimplantation HLA typing in 147 cycles, 109 of which were carried out as part of preimplantation genetic diagnosis (PGD) for Fanconi anaemia, thalassaemia, Wiscott-Aldrich syndrome, hyperimmunoglobulin M syndrome, hypohidrotic ectodermal dysplasia with immune deficiency, and X-linked adrenoleukodystrophy, and 38 for the sole purpose of HLA typing for leukaemias and aplastic and Diamond-Blackfan anaemias. The applied method resulted in the accurate pre-selection and transfer of HLA-matched embryos, yielding 25 clinical pregnancies and the birth of 14 HLA-matched children to the siblings who required stem cell transplantation. PMID- 15354312 TI - International Society of Paediatric Oncology, SIOP XXXVI Congress Meeting. Oslo, Norway, 16-19 September 2004. Abstracts. PMID- 15354313 TI - Validation and application of an automated 96 -well solid-phase extraction liquid chromatography-tandem mass spectrometry method for the quantitative determination of SCH 201781 in human plasma. AB - SCH 201781 is a direct thrombin inhibitor recently under study in clinical trials to determine its safety and efficacy for the treatment of venous and arterial thrombosis. In aqueous solution, SCH 201781 exists as three forms, a ring-opened hydrated form and two ring-closed diastereomers. An automated solid-phase extraction LC-MS/MS method that chromatographically separates and measures each form was developed and validated from 1 to 1000 ng/mL in human plasma. For calibration curve standards, within- and between-run precision (%CV) ranged from 0.6 to 13.7%, while accuracy (%bias) ranged from -4.8 to 13.1%. For quality control samples, within- and between-run %CV ranged from 1.5 to 9.9% while %bias ranged from -9.1 to 4.9%. The method requires a sample volume of 0.8 ml and utilizes 2H6-labeled SCH 201781 as the internal standard. For sample processing, an Isolute C-8 96-well solid phase extraction plate and a Tomtec Quadra 96 sample processor is employed. Separation of the three forms of SCH 201781 is achieved using a 5 microm, 2 mm x 100 mm Asahipak C8 HPLC column and gradient elution. A Sciex API 365 equipped with a turbo ionspray source is used in the selected reaction monitoring mode for detection. The validated method was used to support clinical studies. PMID- 15354314 TI - Black hole of despair swallows up the stars that fell to earth. PMID- 15354315 TI - Hospital eHealth strategy survey 2004. PMID- 15354316 TI - [Report of one caise of renal giant cell malignant fibrous histiocytoma]. PMID- 15354317 TI - Report of the laboratory services committee of the ISBI. PMID- 15354318 TI - Modulatory effect of soy isoflavones on biochemical alterations mediated by TPA in mouse skin model. AB - Exposure to various carcinogenic agents along with other contributing factors increase the risk of cancer formation. The current study assesses the effect of soy isoflavones on the biochemical events associated with tumor promotion in mouse skin. 12-O-tetradecanoyl phorbol-13-acetate (TPA), a well-known tumor promoter on topical application depletes the reduced glutathione content (GSH) and down regulates the activities of its metabolizing enzyme, glutathione-S transferase (GST) and the activities of antioxidant enzymes. However, the ornithine decarboxylase (ODC) activity and unscheduled DNA synthesis are elevated on single topical application of TPA to the dorsal cutaneous portions of the mice. Topical applications of soy isoflavones, half-an-hour prior to the application of TPA prevented the induction of ODC activity and DNA synthesis mediated by TPA (p < 0.01). The content of GSH, GST and antioxidant enzymes (p < 0.05) was also recovered significantly by soy isoflavones in a dose dependent manner. Parallel to these effects, pretreatment with the soy isoflavones also reduced hydrogen peroxide (H2O2) content (p < 0.05) at 1.0 and 2.0 microg/0.2 ml vehicle/animal. Therefore, we conclude that soy isoflavones are potentially protective against TPA induced biochemical alterations. PMID- 15354319 TI - Basic toxicology and metabolism studies of 1,5-anhydro-D-fructose using bacteria, cultured mammalian cells, and rodents. AB - 1,5-Anhydro-D-fructose (AF) is a monosaccharide occurring in edible morels, red seaweeds and certain mammalian tissues. It can be formed directly from starch and glycogen in vivo by alpha-1,4-glucan lyase (EC 4.2.2.13). In this study, the toxicity, absorption and metabolism of AF using bacteria, mammalian cells, rat and mouse models were examined. In Ames test, AF showed no genotoxicity using five strains of the bacterium Salmonella typhimurium TA 98, 100, 102, 1535 and 1537. AF caused no mammalian gene mutation as tested with mouse lymphoma L5178Y cells. AF did not cause toxic symptoms in rats when it was administered as a single oral dose of 5 g/kg and observed over a 14-day period. Furthermore, at necropsy, no signs of abnormality were detected. Daily intraperitoneal (ip) administration of 2 g/kg AF to mice did not induce adverse effects throughout a 28-day period. Radioactive tracing experiments using 14C-labeled AF indicated that AF was efficiently absorbed since the major portion of radioactive material was recovered in urine. Further work using unlabeled AF indicated that the cyclic polyol 1,5-anhydro-D-sorbitol (AS) increased dramatically in both blood and urine upon AF administration at 1 g/kg ip, suggesting the existence of an efficient reduction mechanism from AF to AS, which was then excreted in urine. In conclusion, these studies indicate that AF had low or no toxicity and showed no mutagenicity. PMID- 15354320 TI - Absence of carcinogenic and anticarcinogenic effects of annatto in the rat liver medium-term assay. AB - Annatto (Bixa orellana L.) is a natural food colorant extensively used in many processed foods, especially dairy products. The lower cost of production and the low toxicity, make annatto a very attractive and convenient pigment in substitution to the many synthetic colorants. In the present study we investigate the carcinogenic and anticarcinogenic effects of dietary annatto in Wistar rat liver using the preneoplastic glutathione S-transferase (GST-P) foci and DNA damage biomarkers. Annatto, containing 5% bixin, was administered in the diet at concentrations of 20, 200, and 1000 ppm (0.07; 0.80 and 4.23 bixin/kg body wt/day, respectively), continuously during 2 weeks before, or 8 weeks after DEN treatment (200 mg/kg body wt, i.p.), to evaluate its effect on the liver carcinogenesis medium-term bioassay. The comet assay was used to investigate the modifying potential of annatto on DEN (20 mg/kg body wt)-induced DNA damage. The results showed that annatto was neither genotoxic nor carcinogenic at the highest concentration tested (1000 ppm). No protective effects were also observed in both GST-P foci development and comet assays. In conclusion, in such experimental conditions, annatto shows no hepatocarcinogenic effect or modifying potential against DEN-induced DNA damage and preneoplastic foci in the rat liver. PMID- 15354321 TI - Effect of red pepper Capsicum annuum var. conoides and garlic Allium sativum on plasma lipid levels and cecal microflora in mice fed beef tallow. AB - Antihyperlipidemia or hypocholesterolaemic and antibacterial activities of red hot pepper and garlic are well known. To determine the effect of the dietary spices ingested to suppress blood lipids on the intestinal condition, we examined plasma lipid levels and cecal microflora in mice that were fed diets containing 19% (w/w) beef tallow and 2% red pepper Capsicum annuum var. conoides 'Takanotume' (RP) or garlic Allium sativum 'White' (GP) for 4-weeks. Plasma triacylglyceride level was suppressed by the spices. RP lowered cecal bacteroidaceae, a predominant bacterial group (from 9.4 to 9.0 log CFU/g), bifidobacteria (from 8.7 to 7.6 log CFU/g), and staphylococci. Although GP increased the cecal weight including their contents, significant differences were not shown in the cecal microflora. These results suggest that RP can affect the intestinal condition and host health through the disturbance of intestinal microflora. PMID- 15354322 TI - Safety profile of conjugated linoleic acid in a 12-month trial in obese humans. AB - Conjugated linoleic acid (CLA) is marketed in numerous commercially available dietary supplements, but few studies have looked at the long-term safety of this product. The current study evaluated the safety of one CLA product (Clarinol) over a one-year period in obese humans who were generally healthy. This was a randomized, double-blind study consisting of three phases in which subjects were given 6 g/day of CLA or placebo. Phase 1 was a low calorie diet (13 kcal/kg desirable weight) for 12 weeks or until 10-20% of initial body weight was lost. In phase 2, from weeks 12 to 28, subjects were re-fed a diet providing 25-30 kcal/kg of desirable body weight. Phase 3 was open label, with subjects from both groups taking CLA from weeks 28 to 52. At biweekly visits, subjects completed a questionnaire evaluating side effects and adverse events. Blood was taken for assay of liver function, glucose, insulin, serum lipids, blood counts, and general chemistry. Overall, body composition did not differ between groups. Laboratory tests showed no adverse effects of CLA. Adverse events and side effects were less in the CLA group compared to placebo. We conclude that CLA as Clarinol is safe for use in obese humans for at least one year. PMID- 15354323 TI - Induction of apoptosis in a non-small cell human lung cancer cell line by isothiocyanates is associated with P53 and P21. AB - This study was aimed at examining the effects of glucosinolate derivatives including phenylethyl isothiocyanate (PEITC), benzyl isothiocyanate (BITC), and indole-3-carbinol (I3C), on the induction of apoptosis in human non-small cell lung carcinoma A549 cells. The results indicated that all tested compounds inhibited the growth of A549 cells in a concentration-dependent manner. Flow cytometric analyses and annexin V staining showed that induction of apoptosis occurred at low concentrations of PEITC and BITC (< or = 10 microM), and that necrosis occurred at higher concentrations of PEITC and BITC (25 microM); however, apoptosis was not the major pathway for the antiproliferative effects of I3C. Furthermore, Western blot analyses demonstrated that increased expression of P53 and P21 proteins, but not Bax protein, were associated with PEITC- and BITC induced apoptosis. PMID- 15354324 TI - [A novel coronavirus---SARS virus]. PMID- 15354325 TI - [Progress on the study of epidemiology of severe acute respiratory syndrome]. PMID- 15354326 TI - [Recent advances on the study of mechanism of the effect of lead on hippocampal long-term potentiation]. PMID- 15354327 TI - Prenatal screening for cystic fibrosis: an early report card. PMID- 15354328 TI - In a Vietnamese population, MSX1 variants contribute to cleft lip and palate. AB - PURPOSE: To identify causes of nonsyndromic cleft lip and palate in a Vietnamese population. METHODS: In this study, 175 families with at least one case of cleft lip and/or palate were studied using the candidate genes TGFA, MSX1, and TGFB3. RESULTS: Transmission distortion for alleles of MSX1 were demonstrated for the whole population and two missense mutations were identified, including one (P147Q) that is found in approximately 2% of the population. The P147Q appears to arise from a founder individual based on shared haplotypes in unrelated families. CONCLUSIONS: MSX1 contributes to nonsyndromic clefting in a Vietnamese population, and consistent with other studies, identifiable mutations in this gene cause about 2% of cases of nonsyndromic clefting. PMID- 15354329 TI - Association of CCR5 delta32 deletion with early death in multiple sclerosis. AB - PURPOSE: The interaction between chemokines and their receptors is extremely important in controlling T cell migration into sites of CNS inflammation. Because trafficking of inflammatory T cells into the central nervous system (CNS) is a key player in the pathogenesis of multiple sclerosis (MS), we investigated the possible association of CCR5 delta32 deletion in this disorder. METHODS: DNA isolated from postmortem brain tissue samples of 132 patients with MS and from blood tissue samples of 163 gender and ethnicity-matched healthy controls was used to screen for the CCR5 delta32 deletion allele. RESULTS: An increased frequency of 32-bp deletion allele was found to be associated with early death (P = 0.00005) and with a progressive reduction in the years of survival (onset to death). The death hazard ratio of CCR5 with deletion versus no deletion was 2.12, suggesting that MS patients with the 32-bp deletion have twice the mortality rate of patients with the normal genotype. This effect was more significant in females (hazard ratio 3.58). CONCLUSION: A strong association of the CCR5delta32 deletion with early death could serve as a prognostic marker for MS. PMID- 15354330 TI - FISH analysis helps identify low-level mosaicism in Ullrich-Turner syndrome patients. AB - PURPOSE: To search for X or Y chromosome mosaicism in 45,X individuals using fluorescent in situ hybridization (FISH). METHODS: From our series of 53 Ullrich Turner syndrome patients, we used interphase FISH to evaluate the 19 who had an apparently nonmosaic 45,X karyotype with G-banding. RESULTS: Of those 19 patients, mosaicism was detected in seven (37%), five patients had an XX line, one had a monocentric isochromosome X, and one had a dicentric isochromosome X. No Y chromosome mosaic was identified. CONCLUSION: FISH analysis is a sensitive and cost-effective adjunct to karyotype analysis to identify sex chromosome mosaicism in UTS. PMID- 15354331 TI - Cystic fibrosis screening: lessons learned from the first 320,000 patients. AB - PURPOSE: To examine the data from > 335,000 Cystic fibrosis (CF) tests to detect unsuspected findings and obtain clinical data when indicated to optimize genetic counseling. METHODS: A proprietary database containing 335,204 consecutive CF DNA tests and 445 CF prenatal diagnostic tests was queried. Clinical information was obtained for prenatal and selected nonprenatal cases by telephone contact with physician offices. RESULTS: The mutation 1078delT was found in much lower frequency than expected with rates of only 1:55,867 tests and 0.06% of CF mutations. This level is below the threshold set by the American College of Medical Genetics. Homozygosity was observed for 2789+5G>A in a 29-year-old women and compound heterozygosity with delta F408 in a 40-year-old woman with isolated chronic sinusitis. Many patients elected prenatal diagnosis when not at a 1:4 risk due to echogenic bowel or IVS-8 5T issues. CONCLUSIONS: With the exception of 1078delT, all CF mutations in the ACMG panel were detected with a frequency of > 0.1% of CF chromosomes. When ACMG guidelines are strictly adhered to, population-based CF carrier screening will accurately identify couples at risk for having children with CF. PMID- 15354332 TI - Preconception and prenatal cystic fibrosis carrier screening of African Americans reveals unanticipated frequencies for specific mutations. AB - PURPOSE: It is recommended that cystic fibrosis (CF) carrier screening be made available to African Americans who are either pregnant or planning a pregnancy. We analyzed the carrier and mutant allele frequencies for African Americans undergoing CF carrier screening in our laboratories. METHODS: Between December 2001 and September 2003, we performed carrier screening for 2189 African Americans, testing for at least the 25 recommended mutations. RESULTS: A total of 33 CF carriers were identified. The most common mutations detected were deltaF508, G622D, R117H/7T, and G551D. The G622D allele frequency among African Americans was 0.18%. We did not detect any 3120 + 1G --> A carriers, although 4 were expected (P < 0.05). CONCLUSIONS: When considering only the 25 recommended CF mutations, 1 in 75 African Americans screened in our laboratories were carriers (within the expected range, given a 69% mutation detection rate). The addition of 2 mutations, G622D and Q98R (incidentally identified while screening for ACOG/ACMG mutations), increased the observed carrier frequency to 1 in 66, which is not significantly different from the known African American carrier frequency of 1 in 65. The frequencies of several specific mutations detected were unanticipated, as was the absence of 3120 + 1G --> A carriers. Further studies on African American patients with classic CF are needed to examine the incidence of CF mutations that are not part of the current panel, such as G622D. PMID- 15354333 TI - Molecular screening for diseases frequent in Ashkenazi Jews: lessons learned from more than 100,000 tests performed in a commercial laboratory. AB - PURPOSE: To determine the frequency of carriers of Ashkenazi Jewish (AJ) genetic diseases in the US population and compare these numbers with previously published frequencies reported in smaller more isolated cohorts. METHODS: A database containing more than 100,000 genotyping assays was queried. Assays for 10 separate AJ genetic diseases where comparisons were made with published data. RESULTS: As expected, we observed lower carrier frequencies in a general, US population than those reported in literature. In 2427 patients tested for a panel of 8 AJ diseases, 20 (1:121) were carriers of two diseases and 331 (1:7) were carriers of a single disease. Fifty-three of 7184 (1:306) individuals tested for Gaucher disease had 2 Gaucher Disease mutations indicating a potentially affected phenotype. CONCLUSIONS: As the number of AJ diseases increases, progressively more individuals will be identified as carriers of at least one disease. PMID- 15354334 TI - Family history of heart disease and cardiovascular disease risk-reducing behaviors. AB - BACKGROUND: Family history is an important cardiovascular disease (CVD) risk factor. Preventive behaviors, including lifestyle modifications, can attenuate CVD risk. We studied the association between family history-based heart disease (HD) risk and CVD risk-reducing behaviors. METHODS: Using data from the 2001 Healthstyles survey, we compared frequencies of CVD risk-reducing behaviors among adults without known CVD in categories defined by family history-based HD risk. We classified respondents' HD risk as average (no first-degree relatives with HD), moderate (one relative), or high (> or = two relatives). Behaviors studied included lifestyle modifications, cholesterol measurement, and aspirin use. RESULTS: Of 3383 respondents without known CVD, 28% were classified as being at moderate risk and 15% as being at high risk for HD based on family history. Adjusted odds ratios indicated that moderate- and high-risk respondents were more likely to report having cholesterol measured within the previous 5 years (OR = 1.39, 95% Confidence Interval [CI] = 1.16-1.67 and 1.29, 95% CI = 1.01-1.64, respectively), and aspirin use to reduce CVD risk (OR = 1.49, 95% CI = 1.23-1.79 and 1.67, 95% CI = 1.33-2.09, respectively) than average-risk respondents. CONCLUSION: Almost one half of respondents reported a family history of HD. Aspirin use and cholesterol measurement (i.e., behaviors that health-care providers might suggest) were more likely to be reported by moderate- and high risk respondents than were lifestyle changes. Family history merits further investigation as a public health tool to identify persons with increased HD risk who might benefit from enhanced prevention strategies. PMID- 15354335 TI - Single-nucleotide polymorphisms in SCN5A gene in Chinese Han population and their correlation with cardiac arrhythmias. PMID- 15354336 TI - Magnetic resonance imaging of the intrauterine fetal genitourinary tract: normal anatomy and pathology. AB - Although ultrasonography (US) remains the most widely used diagnostic imaging modality for routine evaluation of the fetus, magnetic resonance (MR) imaging has become an invaluable complement to US in all cases in which additional information is desirable. While the ability of US to detect fetal abnormalities is limited in cases such as maternal obesity, oligohydramnios, and in certain fetal positions, MR using fast and ultrafast pulse sequences enables high-quality fetal images to be acquired regardless of the mother's physical condition or fetal position. Fetal genitourinary disorders are the most common intrauterine abnormalities detected by US, accounting for approximately 30% of all antenatally detected anomalies. Although they usually occur in isolation, these defects can form part of more complex syndromes or chromosomopathies, and MR is indicated to rule out associated abnormalities. In some severe genitourinary disorders, there is a severe deficit of amniotic fluid; these cases are associated with other fetal anomalies such as pulmonary hypoplasia and very poor prognosis. In other cases, the amniotic fluid is not compromised, yet the further detection, localization, and characterization of prenatal disorders will have an impact on postnatal follow-up. This article reviews the role of fetal MR in urogenital tract disorders. PMID- 15354337 TI - Vesicoureteral reflux in young patients: comparison of voiding color Doppler US with echo enhancement versus voiding cystourethrography for diagnosis or exclusion. AB - We assessed the accuracy of voiding color Doppler ultrasonography (US) with echo enhancement for diagnosis or exclusion of vesicoureteral reflux (VUR) versus voiding cystourethrography (VCUG) and evaluated patient tolerance of the echo enhancing agent. One hundred twenty-two patients (ages range, 1 month to 17 years) with 244 ureterorenal units underwent voiding color Doppler US with echo enhancement, which was followed by VCUG on the same day. After US of the urinary tract, the bladder was filled with saline solution via catheter. Then an echo enhancing agent was instilled, and color Doppler US was performed. Reflux was diagnosed when microbubbles appeared in the ureter or the pelvicaliceal system. VUR was detected in 98 units. All grades of reflux were identified. In 13 units (13.3%), grade I reflux diagnosed at voiding color Doppler US was not observed at VCUG. No reflux was detected in 146 units by either method. In 231 (94.2%) of the 244 ureterorenal units, there was concordance between the two methods. The echo enhancing agent was well tolerated by all patients. Voiding color Doppler US with echo enhancement is useful for the diagnosis or exclusion of VUR, being as good as VCUG. Therefore, it may reduce the number of patients exposed to ionizing radiation. PMID- 15354339 TI - Radiologic evaluation in planning surgery of renal tumors. AB - The evolution in diagnostic imaging modalities, mainly in computed tomography (CT) and magnetic resonance imaging (MRI), have made it possible to extend the applications of these techniques from diagnosis to staging and surgical planning. Nowadays, the possibility to present images on different planes with an intrinsic resolution close to that of the original axial sections allows presentation of the kidney on sagittal, coronal, and oblique planes. Three-dimensional reconstructions can be obtained with different methods and have attained excellent image quality. Multidetector spiral CT presently is the best technique for planning surgery, but MRI also enables high-quality images to be obtained if state-of-the-art equipment is available. This update reviews the current status and possibilities of diagnostic imaging modalities in planning surgery of renal tumors. PMID- 15354338 TI - Metanephric adenoma of the kidney: case report. AB - Metanephric adenoma is an extremely rare, epithelial, renal tumor with characteristic histology. This tumor is invariably benign. We report a case of metanephric adenoma diagnosed by ultrasound-guided needle biopsy. PMID- 15354340 TI - Small bowel obstruction secondary to Crohn disease: CT findings. AB - We investigated the computed tomographic (CT) findings in patients with small bowel obstruction (SBO) and Crohn disease (CD). Fourteen patients, seven men and seven women (mean age, 41.3 years), were retrospectively reviewed. All presented with clinical symptoms and signs of SBO. Eleven had a history of CD, whereas three experienced the bowel obstruction as the first manifestation of the disease. On CT, features of complete SBO were seen in nine patients, whereas incomplete obstruction was found in the other five. One patient had CT findings of an adhesive obstruction. The other 13 were diagnosed as having CD-related SBO; a markedly stenotic bowel segment caused the obstruction in one patient, and a thickened-wall small bowel segment with luminal narrowing was evident at the transition zone in the other 12. The mural thickening had a target appearance in seven and homogeneous thickening in the other five. Additional thickened bowel segments were found in five patients and mesenteric involvement was found in 10. Five patients were treated conservatively, and the other nine underwent surgery (one with adhesiolysis only). Resection of the stenotic bowel was performed in six patients and stricturoplasty was done in the other two, with associated intestinal biopsy in one of these two patients. Histopathology revealed findings of active on chronic disease in all. CT is frequently performed for suspected SBO, so radiologists should be aware of the diagnosis of CD, because SBO may be its first manifestation. Alternatively, radiologists can accurately diagnose a CD related obstruction in a patient with known CD and differentiate it from an obstruction due to adhesions. Patient management in these cases, however, is based most often on the clinical condition. PMID- 15354341 TI - Crohn disease: magnetic resonance enteroclysis. PMID- 15354342 TI - Small bowel tuberculosis: enteroclysis findings. AB - We describe the radiologic appearances of small bowel tuberculosis as shown by enteroclysis. A total of 265 patients with proven small bowel tuberculosis seen over a period of more than one decade was evaluated. All patients had positive radiologic findings as shown on enteroclysis examination. Of the 265 patients with small bowel tuberculosis, 174 had isolated small bowel involvement with a normal ileocecal region, whereas 91 had associated noncontiguous involvement of the ileocecal region. The most common radiologic finding was the presence of strictures, noted in 62.7% of cases. Most strictures were short, concentric, and smooth in outline. These strictures were solitary or multiple and located mainly in the jejunum. Other radiologic findings were adhesions (21.8%), ulcerations (9.1%), and diffuse thickening of folds (6.4%). Complications noted were in the form of enteroliths, perforations, and fistulae. The radiographic findings of small bowel tuberculosis, although non-specific, may indicate tuberculosis in a high-risk population. PMID- 15354343 TI - Hepatocellular carcinoma: spectrum of contrast-enhanced gray-scale harmonic sonography findings. AB - Sonography (US) is the most commonly used tool in the assessment of patients with chronic liver disease. Nevertheless, small nodules can be overlooked. Moreover, even if the lesion is readily recognizable, difficulty persists in differentiating hepatocellular carcinoma (HCC) from pre-tumoral, dysplastic stages of nodule evolution. In addition, US is the main tool in guiding percutaneous ablation therapies for HCC and in evaluating patients after treatment, but changes are not specific and difficulties persist in recognizing residual tumor during and after the ablation procedure. Contrast-enhanced, gray scale, harmonic US offers new possibilities in the imaging of liver tumors. The real-time, low-mechanical index technique employed in our institution allows a true dynamic assessment of the tumor during the various phases of contrast medium circulation. This pictorial essay focuses on typical and atypical appearances of HCC when imaged with contrast-specific, real-time US. Findings useful for lesion detection, differential diagnosis, posttreatment assessment, and follow-up especially are illustrated. The potential role of contrast-enhanced US in various clinical settings is emphasized. PMID- 15354344 TI - Focal liver lesions: evaluation with contrast-enhanced ultrasonography. AB - The development of contrast agents for ultrasound has significantly extended its clinical applications, particularly in the detection and characterization of focal liver lesions. Real-time evaluation of the vascular phase currently can be achieved by combining new microbubble-specific methods with second-generation contrast agents. The enhancement pattern in the vascular phase combined with the behavior in the liver-specific late phase allows characterization of the most frequent focal liver lesions, with an accuracy similar to those of dynamic computed tomography and magnetic resonance imaging. PMID- 15354345 TI - MRCP pitfalls. PMID- 15354346 TI - PET/CT molecular imaging in abdominal oncology. PMID- 15354347 TI - PET-CT detection of abdominal recurrence of ovarian cancer: radiologic-surgical correlation. AB - The diagnosis of recurrent ovarian cancer can be difficult on cross-sectional imaging, and variable sensitivities and specificities have been reported for positron emission tomography (PET). Combined functional and anatomic imaging with PET plus computed tomography (CT) potentially allows for improved detection of tumor masses. We investigated the sensitivity, specificity, and accuracy of PET CT for the diagnosis of recurrent ovarian cancer. Sixteen women with previously treated ovarian cancer underwent imaging on a combined PET-CT scanner followed by surgery to assess for possible recurrent disease. The fused PET-CT images were retrospectively reviewed for recurrent disease, and the results of PET-CT were compared with the operative notes. Eleven of the 16 patients had recurrent disease at surgery. The sensitivity, specificity, and accuracy of PET-CT for disease detection on a per-patient basis were 72.7%, 40%, and 62.5%, respectively. For cases of malignant adenopathy (n = 7), 100% were detected on PET-CT. For peritoneal lesions no larger than 1 cm (n = 23), 13% were detected on PET-CT. For peritoneal lesions larger than 1 cm (n = 8), 50% were detected on PET CT. The sensitivity of PET-CT for recurrent ovarian cancer is moderate in patients with low volume disease. A trial involving a larger number of patients with a spectrum of disease volumes is necessary to determine the impact of PET-CT in clinical practice. PMID- 15354348 TI - C20orf9-003 (ACI-1), a gene localized on chromosome 20q13.12 encoding for a 49 kD cytoplasmic protein with a putative nucleotide binding site. AB - Murine NGD5 is a gene identified from NG108-15 cells which is postulated to be involved in opioid receptor function. Here we report the cloning and characterization of a cDNA C20orf9-003 (ACI-1) encoding the human orthologue of the mouse NGD5. Analysis of the genomic structure revealed that C20orf9-003 (ACI 1) contains 13 exons and 12 introns, spanning 52.5kb of genomic DNA and is a variant of C20orf9. Chromosomal localization of human C20orf9-003 (ACI-1) assigned this gene to chromosome 20q13.12. Genes at this locus have been associated with the progression and possibly the development of various cancers. In addition several linkage studies support the possibility that one or more genes affecting obesity are located in 20q13. No function can be clearly assigned to C20orf9-003 (ACI-1), however, the protein has a cytoplasmic subcellular location and the secondary structure contains a Rossman fold like feature which is found in many nucleotide binding proteins. PMID- 15354349 TI - Identification of the human ubiquitin specific protease 31 (USP31) gene: structure, sequence and expression analysis. AB - Parkinson's disease (PD) is a common neurodegenerative disorder with clinical features of bradykinesia, rigidity and resting tremor resulting from the deficiency of dopamine in the nigrostriatal system. Recently, PARK6 was identified as a novel locus associated with autosomal recessive PD. Here we report the identification and characterization of a novel human deubiquitylating gene (USP31), which maps to the critical PARK6 region. Database analysis and 5' RACE identified a 4070bp cDNA, encoded by 27 exons spanning approximately 105kbp of genomic sequence. The predicted protein of 1035 amino acids included a conserved ubiquitin hydrolase region (Prosite profile PS50235), a DUSP (domain in ubiquitin specific proteases-Smart00695) and a ubiquitin-like domain (Prosite pattern PS00299). Northern blot analysis revealed a single USP31 transcript of approximately 4 kb, which was primarily expressed in the testis and lung. PMID- 15354350 TI - Structure of the bovine natural resistance associated macrophage protein (NRAMP 1) gene and identification of a novel polymorphism. AB - The NRAMP 1 gene is a major candidate gene influencing the outcome of infections with intracellular pathogens in numerous species. NRAMP 1 is highly conserved in many mammalian species and the NRAMP 1 gene shows considerable conservation in structure between mice and humans. The association of NRAMP 1 gene polymorphisms with disease in cattle has been limited to a single microsatellite located within the 3'-non coding region of the bovine NRAMP 1 gene. In order to facilitate further studies on this important gene, we now report the nearly complete structure of the bovine NRAMP 1 gene, including sizes and positions of 13 introns relative to the bovine NRAMP 1 gene coding sequence and the DNA sequence of intron-exon junctions. Comparison of the bovine, murine and human NRAMP 1 gene structures revealed a high degree of conservation in intron placement, though the lengths of several introns were less-well conserved. In general, the greatest divergence in intron lengths occurred in regions of the NRAMP 1 gene displaying the lowest coding sequence conservation. In addition, mutations near intron-exon junctions could account for 25 of the 75 total amino acid differences between murine and bovine NRAMP 1. Using information gained through this study, it was possible to rapidly identify a novel polymorphism within the bovine NRAMP 1 gene intron X. This polymorphism was shown by direct DNA sequence analysis to consist of insertion of three guanine nucleotides at positions 37,40 and 98 relative to the intron X start point. Initial scans of several cattle breeds suggest that the two intron X alleles identified here are stable and widespread in the Bos taurus population. PMID- 15354351 TI - Molecular cloning of bovine eIF5A and deoxyhypusine synthase cDNA. AB - Deoxyhypusine synthase is the first of the two enzymes that catalyzes the maturation of eukaryotic initiation factor 5A (eIF5A). The mature eIF5A is the only known protein in eukaryotic cells that contains the unusual amino acid hypusine (N(epsilon)-(4-amino-2(R)-hydroxybutyl)-lysine). Synthesis of hypusine is essential for the function of eIF5A in eukaryotic cell proliferation and survival. Here we describe the cloning and characterization of bovine eIF5A and bovine deoxyhypusine synthase. The deduced bovine eIF5A protein is 100% identical to human eIF5A-1, and the deduced bovine deoxyhypusine synthase protein showed a 93% identity to the human protein. PMID- 15354352 TI - Genomic cloning of a heat shock cognate 71-1 gene (HSC71-1) from the hermaphroditic fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae). AB - The self-fertilizing fish Rivulus marmoratus (R. marmoratus) heat shock cognate 71 (HSC71) gene was cloned and characterized recently (Park et al., 2001). Here, we report the isolation of a homologue of the R. marmoratus HSC71 gene via screening of an R. marmoratus genomic DNA library. A 12,591 bp genomic fragment was sequenced and found to contain a 2844 bp open reading frame that consisted of 8 exons and showed high similarity to the previously reported R. marmoratus HSC71 gene. The two genes differed slightly at exons 5 and 8, and intron 3. On a deduced amino acid sequence level, the two R. marmoratus HSC71 genes were highly similar (89.3% in amino acid residues). In this paper, the author presented a homologous gene (R. marmoratus HSC71-1) similar to R. marmoratus HSC71 gene. PMID- 15354353 TI - Isolation and characterization of two cDNAs encoding translation initiation factor 1A from rice(Oryza sativa L.). AB - The eukaryotic initiation factor 1A(eIF1A) is essential for transferring of the initiator Met-tRNA to 40S ribosomal subunits to form the 40S pre-initiation complex. In present study, we describe the cloning and characterization of two eIF1A genes from rice, which were designated as Oryza sativa eukaryotic initiation factor 1A genes OseIF1A-1, OseIF1A-2, respectively. Both rice elF1As shared high identities in amino acids with eIF1A proteins from other eukaryotes. The mRNA expression analysis revealed that OseIF1A-2 mRNA was much more accumulated than OseIF1A-1 in all tissues but each gene is expressed in root, stem, leaf and flowering spike in high and nearly equal level, and in immature spike in lower level. These results, together with their different location in unrooted phylogenetic tree inferred from amino acid sequences of all known eIF1As, suggested that there are two types of eIF1A genes with different function or different regulation in rice. PMID- 15354354 TI - Isolation and molecular analysis of Umhda2 a gene encoding a histone deacetylase from Ustilago maydis. AB - By use of the polymerase chain reaction and synthetic oligonucleotides designed from conserved regions, we amplified a fragment of a gene from Ustilago maydis encoding a putative histone deacetylase. With this probe we isolated the full gene from a minigenomic library. The gene (designated as Umhda2) contains an open reading frame (ORF) of 1701bp encoding a protein of 566 amino acids. Multiple comparison analysis with other histone deacetylases suggests that the Umhda2 gene product belongs to the Rpd3-related family of proteins. The highest degree of homology with histone deacetylases from other organisms corresponded to Hdalp of Schizosaccharomyces pombe and Rpd3p of Saccharomyces cerevisiae with 64.2 and 62.2% of sequence similarity, respectively. It displayed a substantially lower similarity with another histone deacetylase from U. maydis (Hdalp, 52.4%). Semi quantitative RTPCR results indicate that the gene is transcriptionally up regulated during the in vitro yeast-to-mycelium dimorphic transition. PMID- 15354355 TI - cDNA cloning and characterization of enolase from Chinese cabbage, Brassica campestris ssp. Pekinensis. AB - An enolase-encoding cDNA clone from Chinese cabbage, Brassica campestris ssp. Pekinensis, was isolated. This gene (Accession number: AY307448) had a total length of 1580bp with an open reading frame of 1335bp, and encoded a predicted polypeptide of 444 amino acids with a molecular weight of 47.38 kDa. The deduced amino acid (aa) sequence shared identity with a number of enolases ranging from Bacillus subtilis to human beings and had much higher identity with other plant enolases than with enolases from Bacillus, yeast and human beings. Comparison of its primary structure with those of other enolases revealed the presence of an insertion of 5 amino acids in enolase of Chinese cabbage. Expression of the cloned enolase gene decreased under salt stress, but increased in response to low temperature. Southern blot analysis of genomic DNA indicated that low-copies of enolase gene were present in the genome of Chinese cabbage. PMID- 15354356 TI - Molecular cloning and characterization of GhlecRK, a novel kinase gene with lectin-like domain from Gossypium hirsutum. AB - A novel gene encoding a lectin-like protein kinase was cloned from the upland cotton (Gossypium hirsutum) through cDNA library screening. This gene (named as Ghlecrk; GenBank accession number: AY487461) had a total length of 2233bp with an open reading frame of 1926bp, and encoded a predicted polypeptide of 641 amino acids with a molecular weight of 71.16kDa. The GhLecRK protein shared 73, 65, 64 and 59% identity with other lectin-like kinase proteins isolated from A. thaliana (At3g53810, At2g37710, At3g55550) and Populus nigra (PnLPK) at amino acid level, respectively. Southern blot analysis showed that GhLecRK belonged to a multi-copy gene family. Expression patterns revealed that GhLecRK was enriched in the developing boll (six days post anthesis, 6DPA) and shoot, but low in the root and stem and no expression in the leaf. The domains analysis showed that GhlecRK protein possessed many activating sites/domains including ATP-binding sites, a transmembrane region, a lectin-like domain and a kinase domain. These results indicate that GhlecRK is a lectin-like membrane protein that may play an important role in the phase of fiber development. PMID- 15354357 TI - The Selenophosphate synthetase gene from Leishmania major. AB - Selenophosphate synthetase coding sequence was cloned from Leishmania major by RT PCR amplification. The DNA sequence was found to have an open reading frame encoding protein with 398 amino acids and does not have in-frame UGA codon. The deduced amino acid sequence indicates that it has cysteine residue instead of selenocysteine at the active site of enzyme. Amino acid sequence alignment of Selenophosphate synthetase from parasite with the human enzyme showed approximately 45% homology. The sequences also indicated presence of conserved amino acid residues and motifs that are present in mammalian Selenophosphate synthetase. Southern analysis done with restriction enzyme digested genomic DNA and pulse filed separated chromosome suggests that L. major genome contain a single copy of Selenophosphate synthetase sequence. Expression analysis by Northern analysis and RT-PCR indicated Selenophosphate synthetase mRNA is present in promastigote and amastigote stages of parasite. PMID- 15354358 TI - Isolation and characterization of an IAA-responsive gene from Gossypium barbadense L. AB - The full-length cDNA of an IAA-responsive gene was cloned from Gossypium barbadense L. (designated as Gbiaa-Re) by rapid amplification of cDNA ends (RACE). Gbiaa-Re gene was 1043-bp long and contained a 573-bp open reading frame encoding a polypeptide of 190 amino acid residues. Homology analysis revealed that Gbiaa-Re strongly resembled known plant IAA-responsive genes. The conserved integrated domain "AUX_IAA, AUX/IAA family" resided within the region from L11, to V190 of GbIAA-RE, and the 4 typically conserved domains of IAA-responsive gene family were all found in GbIAA-RE. The secondary structure of GbIAA-RE consisted of 20.53% alpha helix, 13.68% extended strand and 65.79% random coil. In total, 12 phosphorylation sites, 1 N-glycosylation site and 4 O-beta-GlcNAc attachment sites were predicted. Southern blot analysis indicated that Gbiaa-Re belonged to a low-copy gene family. Semi-quantitative PCR analysis indicated that the expression of Gbiaa-Re gene was inducible by IAA. Our studies suggested that Gbiaa-Re was a new member of plant AUX/IAA gene family. PMID- 15354359 TI - The complete coding region sequence of river buffalo (Bubalus bubalis) SRY gene. AB - The Y-linked SRY gene is responsible for testis determination in mammals. Mutations in this gene can lead to XY Gonadal Dysgenesis, an abnormal sexual phenotype described in humans, cattle, horses and river buffalo. We report here the complete river buffalo SRY sequence in order to enable the genetic diagnosis of this disease. The SRY sequence was also used to confirm the evolutionary divergence time between cattle and river buffalo 10 million years ago. PMID- 15354360 TI - [Infantilization: the elderly dependent person is an adult]. PMID- 15354361 TI - [Recreation in nursing homes]. PMID- 15354362 TI - [Doing away with guilt and opening up a dialogue. Interview by Helene Delmotte]. PMID- 15354363 TI - America's crushing medical liability crisis is changing the practice of medicine as we know it. PMID- 15354364 TI - A few tips on writing well. PMID- 15354365 TI - Disability income. PMID- 15354366 TI - Becoming a physician works like this. . . Moving from one life to another. PMID- 15354367 TI - Two bites at the apple: holding physician practices directly liable for medical malpractice. PMID- 15354368 TI - What's "normal": female genital mutilation, psychology, and body image. AB - Despite international efforts to halt the practice of female genital mutilation (FGM), the number of African girls and women undergoing the procedure is not declining as rapidly as international observers had hoped when the World Health Organization began focusing attention on the practice in the 1960s. This article focuses on the psychological effects of FGM through the example of a patient who had undergone the procedure in childhood and now felt that her closed appearance was "normal" and that to be opened would be "abnormal." Western advocates must educate themselves about the various cultural forces that lead to FGM in order to help women who have undergone the procedure heal psychologically, thereby breaking the pattern of abuse from generation to generation. PMID- 15354369 TI - Improving breast health education for Hispanic women. AB - Hispanic women are less likely to receive screening mammograms than are white or African American women, even though breast cancer is the leading cause of cancer related deaths in the population. Between 1990 and 2000, the Hispanic population of Indianapolis, Indiana (Marion County) grew by approximately 300%: from 8450 to 33 290. The present project was undertaken to provide educational outreach in Spanish to the new Hispanic residents of Indianapolis. A bilingual outreach coordinator visited community centers, churches, and health clinics and presented information to Hispanic women and men about a variety of cancers for which Hispanic women are at particularly high risk, including breast and cervical cancer. The number of Hispanic women obtaining mammograms rose by more than 200% during the first 6 months of the program. The authors conclude that culturally appropriate educational outreach presented by a Spanish-speaking woman using a Spanish-language videotape about mammography can improve breast cancer screening in Latinas. PMID- 15354370 TI - A multilevel analysis of women's status and self-rated health in the United States. AB - OBJECTIVES: We investigated associations between various dimensions of women's status and self-rated health. We hypothesized that women living in states with lower levels of women's status would be at greater risk of reporting poor health than women living in states with higher levels of women's status, even when controlling for individual and state characteristics. METHODS: We used individual self-rated health and sociodemographic characteristics for 87 848 female respondents to the Behavioral Risk Factor Surveillance System (BRFSS) 2000. Women's status indices and indicators were obtained from the Status of Women in the States 2000. State median income and income inequality were included as state level contextual variables. We examined data using a multilevel logistic regression method. RESULTS: Women living in states scoring in the lowest quintile on women's political and economic indices (political participation, employment and earnings, and economic autonomy) were more likely to report poor health, odds ratio (OR) 1.14 (95% confidence interval [CI] 1.01, 1.28), OR 1.29 (CI 1.08,1.55); OR 1.30 (CI 1.09, 1.56), respectively, after controlling for individual characteristics and other state-level variables than were women living in the highest-performing 20% of states. CONCLUSIONS: The status of women at the state level has an independent association with the risk of reporting poor health over and above women's individual characteristics. This finding suggests a contextual effect of women's societal status on health status. PMID- 15354371 TI - Antepartum and postpartum depression: healthy mom, healthy baby. AB - This article summarizes the epidemiology, clinical features, and treatment of antepartum and postpartum major depression. This summary will enable primary care clinicians to recognize and screen for perinatal mood disorders and to initiate the risk-benefit decision-making process used in treatment selection. This review will also help identify patient factors that warrant psychiatric consultation. We selected and reviewed studies of antepartum and postpartum depression of greatest utility to primary care clinicians. We developed a graphical algorithm that summarizes the steps needed for the care of postpartum depressed women. The prevalence of postpartum depression ranges from 10% to 15%, with screening rates of depressive symptoms as high as 35% in African American women. The prevalence of antepartum depression is as high as 26% among women in poor, urban communities. Maternal depression is associated with adverse effects on fetal and infant development. The challenge of diagnosing perinatal major depression can be addressed by assessing for persistent cognitive and affective symptoms and functional impairment. Effective treatments include interpersonal psychotherapy and selective serotonin reuptake inhibitors. The treatment of perinatal depression requires a unique informed consent process that is guided by a discussion of maternal preferences, the severity of maternal illness and recommended treatment, the risks of psychotropic exposure, and the risks of untreated mental illness. Rapid and effective mental health intervention in depressed expectant or new mothers not only restores maternal function, but can also prevent adverse infant neurobehavioral outcomes. PMID- 15354372 TI - Comprehensive care for women veterans: indicators of dual use of VA and non-VA providers. AB - OBJECTIVE: To compare women who use Department of Veterans Affairs (VA) health care providers only with women who use VA and non-VA providers (dual users). METHODS: An anonymous survey was mailed to women veterans randomly sampled from 10 VA medical centers in 3 states. We measured reports of dual use of VA and non VA providers according to the VA provider's gender, provision of routine gynecological care by VA provider, use of VA women's clinic, and overall satisfaction with VA care. Multiple logistic regression analyses were performed with adjustment for patient demographics, health status, VA service connection status, and clustering by site to determine what factors influenced dual use of providers. RESULTS: In the fully adjusted logistic regression model, provision of routine gynecological care by VA providers (odds ratios [OR] 0.37; 95% confidence intervals [CI] 0.22, 0.60) and use of VA women's clinics (OR 0.56; CI 0.35, 0.90) were strongly associated with a lower likelihood of dual use. Dissatisfaction with care (OR 1.88; CI 1.04,3.41) and higher income (OR 1.89; CI 1.32, 2.71) were also associated with an increased likelihood of dual use. Having a female VA provider was not associated with dual use. CONCLUSIONS: Women veterans' use of VA and non-VA providers is influenced by the scope of clinical services and dissatisfaction with those services. VA clinics should either promote routine gynecological care within primary care clinic settings or pair traditional primary care with VA women's clinics to enhance coordination and comprehensiveness and, thus, reduce fragmentation of care for veteran women. PMID- 15354373 TI - A psychosocial understanding of depression in women: for the primary care physician. AB - This paper has two basic purposes. First, it will provide an understanding of the risks of depression in women, with a focus on the psychosocial factors. The lifetime prevalence of major depression in women ranges from 10% to 23%, twice the rate among men; until puberty, however, rates are similar among boys and girls. The emergence of sex differences at puberty has generated a great deal of research into the biological (in particular, reproductive hormones) and psychosocial factors that may account for this difference and contribute to depression in women across the life cycle. Depression is associated with significant impairment and physical conditions and, thus, clearly constitutes a significant public health problem. The second goal of this paper is to describe some tools for screening depression in the primary care setting. Ten percent to 25% of patients seen by primary care providers have some type of depressive disorder that affects not only their functioning and well-being, but also their physical health. Providers have limited time and expertise for assessing and treating depression. Information about the context of women's lives and an efficient and brief depression screen can help providers evaluate the presence, severity, and impact of depression in their patients. PMID- 15354374 TI - Innovations in adolescent reproductive and sexual health education in Santiago de Chile: effects of physician leadership and direct service. AB - Reproductive and sexual health (RSH) education is a key component of most family planning programs around the world and is particularly important for adolescents, for whom parenthood is more likely to have difficult or dangerous health outcomes. A lack of comprehensive RSH education targeted at adolescents may augment the poor outcomes associated with early pregnancy by creating barriers to optimal care. This article discusses the creation of the Centro de Medicina Reproductiva y Desarrollo Integral de la Adolescencia clinic, a comprehensive adolescent reproductive health center in Santiago de Chile, and its RSH education programs. In particular, the role of the physician in originating and leading the RSH education efforts, the controversy associated with RSH education in Chile, and the effects of comprehensive RHS education on the local and regional adolescent populations are discussed. PMID- 15354375 TI - The best is yet to be: preventing, detecting, and treating depression in older women. AB - To live fully for as long as life lasts is our goal for aging and the challenge we face as we age ourselves and help our patients age. Although we cannot prevent depression, we have information about the correlates of depression or its absence. There are nonmedical interventions that significantly lessen depressive symptoms. We have data on successful aging in the community that suggest some preventative measures. The article reviews information about the risk factors for depression in older women, the prevalence of depression in older women, and the concept of minor or subsyndromal depression. Finally, current concepts of treatment and data on longitudinal outcomes are presented. PMID- 15354376 TI - Selected integrative medicine treatments for depression: considerations for women. AB - This review evaluates the research published between 1966 and 2004 on several integrative treatments for depression, including omega-3 fatty acids, Hypericum perforatum (St. John's Wort), S-adenosyl-methionine, folate, 5-Hydroxytryptophan, acupuncture, exercise, and light therapy, with a particular emphasis on issues pertinent to women. Data from double-blind, placebo-controlled trials support each of these as treatment interventions for depression. We discuss both the strength of the evidence for each treatment and methodological issues related to interpretation of efficacy. Available data pertaining to considerations in women, including use during pregnancy and breastfeeding and interactions with hormonal therapies are discussed. The reviewed treatments deserve further research. Their appropriate place in the armamentarium of depression treatments for women must be defined. An evidence-based integrative medicine approach brings together treatment options with proven efficacy and the public's desire for complementary and alternative medicine treatments. PMID- 15354377 TI - Do women possess a unique susceptibility to the neurotoxic effects of alcohol? AB - Neurodegeneration and neurological impairment associated with alcohol dependence have been observed predominantly in alcohol-dependent men. Thus little research has examined the neuropathological consequences of alcoholism in women. Recent evidence obtained from both human and animal studies, however, suggests that women may well be at greater risk of alcohol-induced brain injury than men are. Further, researchers have used animal models to identify molecular events that may contribute to this putative sex difference. Such data are likely of significance in both understanding women's unique susceptibility to neurotoxic effects of alcohol and addressing the possibility that the medical management of alcoholism in women and men may require distinct approaches. As a whole, these findings clearly indicate the need for a significant expansion of both clinical and basic science research of this topic. PMID- 15354378 TI - Dental CRM. PMID- 15354379 TI - Infant dental care: changing the mindset of a profession. PMID- 15354380 TI - Providing oral health to the little ones. AB - The dental profession has achieved successes in reducing the incidence of tooth decay and periodontal disease in adults and teens. The same cannot be said of Early Childhood Caries, which is the most prevalent chronic childhood disease and the greatest unmet healthcare need among youngsters, particularly those from underserved populations. The authors elucidate the infectious, transmissible disease process underlying ECC, the milestones at which preventive intervention is vital to successful treatment of infant patients, and protocols for preventive treatment. The concept of the "Dental Home," its critical role in the dental health of families with young children, and the best-practice timeline for its establishment, are delineated. The authors offer guidelines for caries risk assessment, specific treatment recommendations for the prevention of infant caries, and strategies to facilitate pediatric practice. PMID- 15354381 TI - Private practice pediatric dentists in Indiana: location and longevity. PMID- 15354383 TI - Keeping abreast of laws that affect you. PMID- 15354382 TI - Comparing infant oral glucose clearance with and without a water rinse. AB - The purpose of this study was to test the hypothesis that a small rinse of water in an infant's mouth immediately following formula consumption would significantly improve oral glucose clearance. Though statistically significant results were not obtained at all time intervals, this study lends validity to the hypothesis that a small water rinse immediately following formula feeding could help enhance oral glucose clearance in infants. PMID- 15354384 TI - Dopaminergic neurotoxicity of homocysteine and its derivatives in primary mesencephalic cultures. AB - Levodopa and dopamine are metabolized to 3-O-methyldopa and 3-methoxytyramine, respectively, by the enzyme catechol-O-methyltransferase (COMT) leading to the production of the demethylated cofactor S-adenosylhomo-cysteine (SAH) and subsequently homocysteine (HC). Indeed, treatment of Parkinson's disease (PD) patients with levodopa leads to increased HC blood levels. Therefore, HC is discussed to be involved in the pathogenesis of PD as well as in enhanced progression of PD in patients treated with levodopa. Here we investigated the toxicity of HC and its derivatives SAH, homocysteic acid (HCA) and cysteic acid (CA) on tyrosine hydroxylase (TH)-positive neurons in primary mesencephalic cultures from rat in vitro. Furthermore, we evaluated the toxicity of HC on cultures stressed with the dopaminergic neurotoxin 1-methyl-4-phenylpyridinium (MPP+). Incubation with HC or HCA did not result in significant effects on TH positive neuron survival with concentrations up to 1 mM, but led to morphological changes of TH-positive cells with significantly fewer and shorter neurites at concentrations of > or = 100 microM after 48 h. In contrast, SAH and CA were toxic at concentrations of >100 microM after 48h. Furthermore, MPP+ showed strong toxicity towards TH-positive cells after 48 h (half-maximal toxic concentration: 20 microM), whereas co-incubation with HC for 24 or 48 h did not further alter TH positive cell survival. Taken together, our results do not demonstrate relevant dopaminergic toxicity of HC in vitro, and therefore HC is most likely not involved in the pathogenesis of PD or in accelerating the progression of PD by levodopa. PMID- 15354385 TI - MTHFR C677T polymorphism, folic acid and hyperhomocysteinemia in levodopa treated patients with Parkinson's disease. AB - Certain mutations (TT homozygous; CT heterozygous; CC wild-type) of the methylenetetrahydrofolate (MTHFR) gene and long-term levodopa application in patients with Parkinson's disease (PD) support onset of hyperhomocysteinemia. Total plasma homocysteine (t-hcys) depends on B6, B12, folic acid, all of which support remyelination from t-hcys to methionine. Objective of this trial were to compare B6, B12, folic acid and t-hcys levels in plasma of 83 levodopa treated PD patients and 44 controls. PD patients with the CT or TT genotype had significant higher t-hcys levels than controls or PD patients with the CC allele. Concentrations of B6 or B12 did not differ, but folic acid was significant higher in PD patients with the CT mutation. We recommend MTHFR genotyping, t-hcys monitoring and early vitamin supplementation in PD patients. The folic acid increase in PD patients with the CT allele is hypothetically due to an endogenous upregulation of folic acid absorption to decrease t-hcys. PMID- 15354386 TI - Screening for mutations in synaptotagmin XI in Parkinson's disease. AB - Parkinson's disease (PD) is characterized by selective degeneration of neurons in the substantia nigra and subsequent dysfunction of dopaminergic neurotransmission. Genes identified in familial forms of PD encode proteins that are linked to the ubiquitin-proteasome system indicating the pathogenic relevance of disturbed protein degradation in PD. Some of them, i.e. alpha-synuclein, parkin and synphilin-1, have been implicated in presynaptic neurotransmission based on their localization in synaptic vesicles. Synaptotagmin XI is linked to the pathogenesis of PD based on its identification as a substrate of the ubiquitin-E3-ligase parkin. Moreover synaptotagmin XI is involved in the maintainance of synaptic function and represents a component of Lewy bodies (LB) in brains of PD patients. Therefore, we performed a detailed mutation analysis of the synaptotagmin XI gene in a large sample of 393 familial and sporadic PD patients. We did not find any disease causing mutations arguing against a major role of mutations in the synaptotagmin XI gene in the pathogenesis of PD. PMID- 15354387 TI - Depletion and restoration of endogenous monoamines affects beta-CIT binding to serotonin but not dopamine transporters in non-human primates. AB - The radioligand [123I]beta-CIT binds to dopamine transporters in striatum and to serotonin transporters in brainstem. Endogenous dopamine or serotonin may compete with radioligand binding at monoamine transporters. We used alpha-methyl-p tyrosine (AMPT) to block dopamine production and measured [123I]beta-CIT binding before and after endogenous dopamine was restored by IV administration of the dopamine precursor L-dihydroxyphenylalanine (L-DOPA) in rhesus monkeys. P chlorophenylalanine (pCPA) was used to inhibit serotonin production, and [123I]beta-CIT binding was assessed before and after IV administration of the serotonin precursor 5-hydroxy-L-tryptophan (L-5-HTP) restored endogenous serotonin. Pretreatment with benserazide blocked peripheral decarboxylization in both paradigms. Serotonin restoration measurably displaced [123I]beta-CIT binding to brainstem serotonin transporters but not to striatal dopamine transporters. Restoration of dopamine apparently did not affect [123I] beta-CIT binding to striatal dopamine transporters. However, dopamine restoration reduced radioligand binding to brainstem serotonin transporters, most likely due to dopamine release from serotonin neurons following L-DOPA administration. The higher striatal density of dopamine transporters relative to dopamine concentrations may explain why [123I] beta-CIT displacement by endogenous dopamine was not observed. This study indicates that [123I]beta-CIT binding in brainstem (raphe area) is affected by endogenous serotonin release in vivo and that L-DOPA treatment may cause serotonin neurons in the brainstem to corelease dopamine. PMID- 15354388 TI - Differential executive control impairments in early Parkinson's disease. AB - Investigations concerning cognitive functions in early PD have revealed memory and executive function deficits related to dysfunction of fronto-striatal circuitry. Despite the range of data base, many previous investigations are limited because of methodological questions and inconsistencies. Thus the pattern of executive function impairments in early PD is far from being established. In the present investigation, twenty PD patients in early stages of the disease were compared to control subjects on a comprehensive neuropsychological test battery, aiming to explore their cognitive profile across a range of executive subcomponents. Results revealed impairments with respect to initiation, reasoning and planning. In summary, the present investigation shows that PD is associated with a differential executive impairment pattern which is (partly) related to disease characteristics and affective variables. PMID- 15354389 TI - Simple movement sequences better correlate to levodopa plasma levels than complex ones. AB - Instrumental assessment of movements with a computer based device reflect the clinical response of patients with Parkinson's disease (PD) to dopaminergic stimulation. We investigated associations between levodopa plasma levels, scored motor symptoms of upper limbs and instrumental test outcomes after dopaminergic stimulation. Clinical rating scores, test outcomes for simple and complex motion series significantly improved after oral application of 250 mg of a water soluble, fast absorbed levodopa/benserazide preparation, which induced a significant increase of levodopa in plasma during a two hour interval. There was a significant association between the computed area under the curve-values of levodopa plasma concentrations and test results for simple-, but not for complex movement sequences. Performance of complex motion series additionally ask for concomitant cognitive efforts with consecutive hypothetical involvement of extranigral non dopaminergic systems. In contrast, practice of simple movements is more directly associated to the predominantly dopamine regulated motor system. PMID- 15354390 TI - Clinical effects of repetitive transcranial magnetic stimulation versus acute levodopa challenge in Parkinson's disease. AB - We studied the short-term clinical effects of 10-Hz repetitive transcranial magnetic stimulation (rTMS) of the motor hand area contralateral to the more affected limb in 12 non-fluctuating, for at least 12 hours drug free patients with Parkinson's disease (PD). We investigated the efficacy of rTMS in combination with a levodopa challenge test design under double-blind, placebo controlled conditions. Significant reductions of UPDRS III motor scores showed the treatment conditions: placebo/rTMS, levodopa/sham stimulation and levodopa/rTMS. A more detailed evaluation of arm symptoms contralateral to the stimulated brain region showed even more pronounced effects for the three conditions. There were significant differences between the mean response of the UPDRS III arm scores to the four test conditions. In conclusion our study demonstrates short-term beneficial effects of 10-Hz rTMS on motor symptoms in PD patients. A release of endogenous dopamine in subcortical structures, i.e. putamen, in response to rTMS is the most likely mechanism of action. PMID- 15354391 TI - Impact of apomorphine on BOLD signal during movement in normals. AB - Our group investigated modulatory effects of apomorphine on cerebral activation patterns during finger tapping movements in six healthy right-handed volunteers using an established fMRI protocol. Apomorphine application disclosed a reduction of cerebral activation to the contralateral precentral and postcentral gyrus and ipsilateral cerebellum, with a prominent net reduction of BOLD signal in cerebellar areas. These findings contradict those of similar studies performed on dopaminergic function and Parkinson's disease (PD), which predominantly found augmentation of cerebral activation patterns in normal volunteers and PD patients after dopaminergic stimulation. One conceivable explanation for our singular results would be preferred binding of apomorphine to presynaptic dopaminergic receptors, leading to inhibition of endogenous dopamine release and resultant diminished dopaminergic stimulation, reflected in diminished cerebral activation patterns. These findings warrant future consideration and further investigation of possible central inhibitory effects of dopaminergic therapy in functional imaging studies of the dopaminergic system in general and PD in particular. PMID- 15354392 TI - Worsened motor test performance following acute apomorphine injection in previously untreated patients with Parkinson's disease. AB - Instrumental tests and clinical rating assess motor disability in Parkinson's disease (PD) patients. Previous long-term dopaminergic substitution influences the behaviour following acute dopaminergic stimulation. Objective of this study was to investigate the motor response following an apomorphine application in previously untreated-, treated- and before treated PD patients, who received placebo. Outcomes of instrumental tests worsened in previously untreated-, but not in before treated PD patients after apomorphine injection and in the PD subjects under the placebo condition. Generally, rating scores of motor symptoms significantly improved after apomorphine administration, whereas placebo application showed no effects. Tolerance to sedative effects of apomorphine in treated PD patients or sensitivity of employed motor tests to presynaptic dopaminergic autoreceptor mediated inhibition of endogenous dopamine release or postsynaptic dopaminergic overstimulation with resulting decreased cognitive function in previously untreated PD patients hypothetically caused this discrepancy between outcomes of subjective clinical rating and objective motor test performance. PMID- 15354393 TI - Transdermal lisuride delivery in the treatment of Parkinson's disease. AB - Transdermal delivery of dopamine agonists (DA) is a promising therapeutic concept, which aims to ameliorate frequency and intensity of motor fluctuations in patients with Parkinson's disease (PD). We treated 8 PD patients with unpredictable on-off phenomena with lisuride patches (release: 2-5 microg lisuride base/cm2/hour in mice) in addition to their preexisting antiparkinsonian drug regime up to a period of 8 days. In order to quantify the intensity and frequency of motor fluctuations, we determined the motor changing rate (MCR), which corresponds to the patient's self rating of motor function, performed every thirty minutes, divided through the number of scored intervals minus 1. Additional lisuride patch application significantly (p = 0.023) improved the MCR compared to baseline. Relevant side effects were transient skin irritations in four patients. Our observational study demonstrates the safety, tolerability and efficacy of transdermal lisuride delivery in the treatment of motor complications. PMID- 15354394 TI - Congruent deterioration of complex and simple movements in patients with Huntington's disease. AB - Rating scales and assessment of simple and complex movements may reflect severity of Huntington's disease (HD). Objectives of our study were to compare scored HD symptoms and outcomes of instrumental tests, which demand for simple (tapping) and complex (peg insertion) movement series, in controls and subjects in various HD stages and to correlate them to each other. Motor test outcomes were significantly worsened in previously untreated and treated HD patients in comparison with HD gene carriers and controls. Peg insertion- and tapping results significantly correlated with the scored HD symptoms. Significant associations appeared between both motor test results in the controls, the previously untreated- and treated HD patients. Results of both instrumental tasks represent no specific diagnostic marker of HD, but the significant associations between both motor test outcomes indicate, that a parallel progress of deterioration of complex and simple movement abilities occurs after start of HD. PMID- 15354395 TI - Methyltetrahydrofolate reductase polymorphism influences onset of Huntington's disease. AB - Onset of Huntington's disease (HD) negatively correlates with CAG repeat length of the HD gene, which encodes the protein huntingtin. This protein interacts with the homocysteine metabolizing enzyme cystathionine betasynthase (CBS). Objective of this study was to analyze the impact of CAG repeats, polymorphisms of various homocysteine metabolizing enzymes, like CBS, Methyltetrahydrofolate Reductase (MTHTR), Methionine Synthase Reductase (MSR) and methionine synthase (MS) on HD onset in 171 patients. The significant impact of CAG repeats on HD onset (chi2= 25.54, FG = 4, p<0.0001) with a significant correlation between both (R= -0.521, p=0.01) was obvious. HD patients with the homozygous MTHFR-1298-CC significantly (p = 0.024) earlier experienced HD symptoms. There was no influence demonstrable of CBS, MSR and MS. Determination of MTHFR polymorphisms and CAG repeats enables screening for subjects with putative early HD onset in order to study neuroprotective compounds in their efficacy to delay HD symptoms. PMID- 15354396 TI - Coenzyme Q10 serum levels in Huntington's disease. AB - Mitochondrial dysfunction contributes to the neurodegenerative process in Huntington's disease (HD). Coenzyme Q10 (CoQ10) enhances mitochondrial complex I activity and may therefore provide a therapeutic benefit in HD. We compared serum CoQ10 levels of previously untreated-and treated HD patients with those of healthy controls. CoQ10 did not significantly (ANCOVA F(dF 2, dF 55) = 2.57; p=0.086) differ between all three groups. However, the post hoc analysis showed no significant (p = 0.4) difference between treated HD patients ([CoQ10]: 88.12 [mean]+/-24.44 [SD], [range] 48.75-146.32 [pg/million platelets]) and controls (93.71+/-20.72, 65.31-157.94), however previously untreated HD patients (70.10+/ 21.12, 38.67-106.14) had marked (p = 0.051) lower CoQ10 results than treated HD patients and controls (p = 0.017). Our results support that CoQ10 supplementation in HD patients may reduce impaired mitochondrial function in HD. PMID- 15354397 TI - The N-methyl-D-aspartate antagonist memantine retards progression of Huntington's disease. AB - According to the excitotoxicity hypothesis, neurotoxicity due to glutamate is regarded as potential factor in the progredient neurodegeneration of Huntington's disease (HD). Memantine, as a glutamate receptor antagonist, should counteract this mechanism. Its effectiveness (up to 30 mg/day) with regard to retardation of progression was thus examined in 27 HD patients in a two year, open and multicentre trial. The results suggest that memantine treatment of HD may be useful in terms of retardation of the progression of the disorder. PMID- 15354398 TI - Transcranial sonography in spinocerebellar ataxia type 3. AB - BACKGROUND: Transcranial sonography (TCS) identifies hyperechogenic basal ganglia in extrapyramidal disorders such as Parkinson's disease or dystonia and reveals brain atrophy reflecting the stage of degeneration in Huntington's disease. Aim of the present study was to evaluate the diagnostic potential of TCS in spinocerebellar ataxia type 3 (SCA3), a neurodegenerative disease affecting the cerebellum, multiple pontine nuclei, substantia nigra, pallidum, putamen, caudate nucleus and long spinal tracts. METHODS: 15 patients with a molecular diagnosis of SCA3 and 15 age- and sex-matched healthy control individuals were examined with TCS. Echogenicity of the substantia nigra, caudate nucleus, pallidum, putamen, dentate nucleus and cerebellar white matter were determined quantitatively (substantia nigra by measuring the area of incresed echogenicity) or semiquantitatively (0 = none, 1 = mild, 2 = marked) on both sides and compared with the echotexture of defined brain structures. Additionally, the width of the lateral ventricles, the 3rd and 4th ventricle was measured in both groups. RESULTS: SCA3 patients exhibited hyperechogenicities of the cerebellar white matter (57%), substantia nigra (40%), the dentate nucleus (54%), putamen (40%) and pallidum (40%) significantly more frequently (p <0.05) than controls (20%, 13%, 9%, 0%, and 0% in the corresponding areas). In none of the healthy individuals a marked signal increase could be observed, whereas 53% of SCA3 subjects had at least one region with marked echogenicity. Cerebellar atrophy as demonstrated by an enlarged 4th ventricle was observed in all SCA3 patients whereas this structure could not be differentiated from surrounding parenchyma due to its small size in healthy individuals. 3rd and lateral ventricles were significantly larger in SCA patients as compared to controls (p < 0.05). CONCLUSIONS: TCS is a suitable and non-invasive bed-side method to detect basal ganglia hyperechogenic lesions and posterior fossa abnormalities in SCA3 patients. The pattern of echotexture alterations and brain atrophy most likely reflects distribution and morphology of the neurodegenerative process. PMID- 15354399 TI - Therapeutic strategies in Friedreich's ataxia. AB - Friedreich's ataxia is caused by a pronounced lack of frataxin, a mitochondrial protein of not fully understood function. Lack of frataxin homologues in yeast and mice leads to increased sensitivity to oxidative stress, depletion of proteins with iron-sulfur clusters like respiratory chain complexes I-III and aconitase, and to iron accumulation in mitochondria. Similar effects have been demonstrated in human disease with increased markers of oxidative DNA damage in urine and impaired oxidative phosphorylation in in vivo exercise studies using 31 Phosphorus magnetic resonance spectroscopy (31P-MRS). Therapeutical trials mainly focus on antioxidative treatment with coenzyme Q10 or its short-chain variant idebenone. Promising effects on cardiac hypertrophy in uncontrolled preliminary studies contrast with minor effects in controlled trials and no effect of antioxidants on neurological deficits has been established. Preliminary encouraging 31P-MRS data exist for the treatment with L-carnitine but not with creatine. However, all these interventions may take effect too late in the pathogenic process. Alternative strategies aiming at an enhancement of frataxin by stem cell transplantation, gene transfer or frataxin supplementation are desirable. Additionally, more efficient biomarkers are needed to monitor treatment effects. PMID- 15354400 TI - Cholecystokinin- and cholecystokinin-B-receptor gene polymorphisms in panic disorder. AB - Panic disorder like other neuropsychiatric disorders is believed to be caused by multiple psychosocial and biological factors. Several lines of evidence point to a role for the peptide neurotransmitter cholecystokinin in the pathogenesis of panic disorder. We therefore determined the allele and genotype frequencies of a single nucleotide polymorphism in the CCK gene (-36C>T) and one CT repeat polymorphism in the CCK-B-receptor gene in a German panic disorder sample (n = 115 for CCK gene polymorphism, n = 111 for CCK-B-receptor polymorphism) and compared them with gender and age matched controls. The length of the polymorphic CT repeat alleles varies between 146 bp and 180 bp. We first analysed the results by a permutation test which provided evidence for heterogeneity between patients and controls (p=0.002). We then analysed the data as a di-allelic polymorphism with a short (146-162bp) and a long (164-180bp) allele and as a tetra-allelic polymorphism with 4 alleles (146-154bp, 156-162bp, 164-170bp, 172-180bp). In the di-allelic analysis as well as in the tetra-allelic analysis there was an excess of the longer allele (p = 0.001) or the two longer alleles (p = 0.041) respectively in patients with panic disorder. No difference between groups was observed for the -36C > T polymorphism. Our findings are consistent with the notion that genetic variation in the CCK neurotransmitter system contributes to the pathogenesis of panic disorder. PMID- 15354401 TI - Pet models in cancer research: general principles. AB - Oncology has made great advancements in the past 50 years, moving from preliminary to complex studies and developing in the process numerous models. An important function in this development has been played by animal investigations that have displayed many aspects of cancer and led to the discovery of new therapies. Nevertheless, the debate about preclinical "tools" suited to predict efficacy as well as side effects of anticancer compounds and treatments is open. In this review we focus on the role of pet models in cancer research, whose continuously increasing importance is due to the disclosure of striking histopathological, anatomical, genetical, and biomolecular similarities among feline, canine, and human tumors. Remarkably, the improvement of clinical condition of companion animals, obtained by their enrolment in cancer trials, is generally perceived as an added value for the whole society. In the first paragraphs we examine crucial ethical, clinical, and financial issues that make up the framework of this area of translational research. Then we illustrate the new figures of researchers, namely experts in laboratory-clinic interface, who are needed in this field, and describe the relevant potentialities of pet cancer registries and genome projects. In the conclusions are summarized the principal arguments that support the adoption of pet models in tumor studies. PMID- 15354402 TI - Immunodeficiency in different histotypes of radically operable gastrointestinal cancers. AB - Cell-mediated immunodeficiency, with Total and T lymphocytes count decrease, is well established in cancer patients and it predicts a poor prognosis and poor survival rates. Furthermore, major surgery induces a transient immunodeficiency, too. Nevertheless, cell-mediated immunity in pancreatic cancer, which has a very poor prognosis, has not been completely outlined. Aim of this study is to evaluate the cell-mediated IL-2 dependent immune status in operable pancreatic cancer patients and to compare it with other gastrointestinal tumors. One hundred and twenty-one cancer patients (22 pancreatic, 48 gastric and 51 colorectal), with a median age of 66 years (range 42-83), 55 males and 66 females, were enrolled. Total lymphocyte count and lymphocytes subset (T helper count - CD4+) were assessed preoperatively and on the 14th and 50th postoperative day. Results obtained were compared between the groups and related to nodal involvement (N0 versus N+). Colorectal and gastric cancer patients showed quantitative lymphocyte deficiency at baseline in 29% and 41% of cases, respectively. Fourteen days after surgery values below normal range were found in 44% and 54% (Total) and 53% and 67% (T helper), respectively. Recovery of postoperative surgery-related lymphocytopenia occurred late only in patients with normal count at baseline. According to regional nodal involvement (pN0/N+) T helper deficiency was significantly more frequent in patients with nodal involvement than in patients without. In pancreatic cancer, percentage of immunodepressed patients at baseline was higher compared to the other two groups (71%). Lymphocyte count was significantly different between pancreatic and gastric/colorectal cancer, reaching a statistical significance at baseline and on the 14th and 50th postoperative day. No differences of T helper deficiency were noted according to nodal involvement (N0 versus N+) neither at baseline nor in the postoperative period. In conclusion, the degree of immunosuppression varies among different tumor types: since initial stages of disease, immunodepression was significantly greater in pancreatic cancer which should be considered always a systemic disease even in early stages and indipendently from the nodal involvement and from tumor load. PMID- 15354403 TI - Spinal cord ependymomas in adults: analysis of 15 cases. AB - This retrospective analysis was performed to examine the outcome of patients with spinal cord ependymomas treated with surgery and postoperative radiation therapy between 1982 and 1998. There were 10 male and 5 female patients, ranging from 16 to 74 years of age with a median age of 38 years. Surgery was gross total resection in 2 patients, subtotal resection in 10, biopsy in 3. All patients received radiation therapy with a total dose of 40-56 Gy. The 5 and 10 year overall survival rates were 83.3 and 83.3%, respectively. Twelve patients are still alive at a median follow-up period of 70 months. Of the 15 patients, 6 developed recurrent disease on follow-up. The median time to recurrence was 45 months (range: 24-80 months). Local failure within the initial irradiated volume occurred in 3 out of 6 patients who received less than 45 Gy and 2 out of 8 patients treated with more than 45 Gy. Four out of the six failures were salvaged with additional treatment. Re-irradiation was used as a part of salvage or sole treatment in 3 cases. The patient who was salvaged with radiation therapy only died of disease progression 41 months following recurrence and the other two who received a combination of surgery, radiotherapy or chemotherapy were still alive 57 and 30 months following relapse. The present study shows that surgery and post operative radiation treatment for spinal ependymoma patients resulted in high survival rates. Patients with residual disease after surgery should be treated with radiation therapy with a dose of more than 45 Gy. Re-irradiation may be the treatment of choice for recurrent patients having less than complete resection or no surgery. PMID- 15354404 TI - Single-agent vinorelbine in pretreated breast cancer patients: comparison of two different schedules. AB - This retrospective study compared toxicity and activity of vinorelbine according to two schedules with different projected dose intensities in heavily pretreated breast cancer patients. Forty patients were assessable for toxicity and activity in each group; group A received vinorelbine 25 mg/m2 week + lenograstim (150 microg/m2 s.c. on day 3); group B received 25 mg/m2 on days 1 and 8 every 3 weeks. The projected dose intensity was 25 mg/m2/week and 16.6 mg/m2/week, and delivered dose intensity 95.2% and 94.5% in group A and B, respectively. Grade 3 4 afebrile neutropenia was recorded in 25% and 37.5% of patients in A and B, respectively. Overall response rate, 52.5% and 35%; no change, 35% and 40%; progression of disease, 12.5% and 25% in A and B, respectively. Median duration of the response was 10 months for group A and 7 months for B. Median time to progression: 9.0 months and 4.0 months for A and B, respectively. At a median follow-up of 45 months for group A and 19 months for group B, median overall survival was 19 months and 16, respectively. In conclusion the results of the study showed that dose intensity of vinorelbine could have an improvement in terms of time to progression in pretreated advanced breast cancer. PMID- 15354405 TI - Lymphadenectomy in gastric cancer: influence on prognosis of lymph node count. AB - To better understand the role of the number of lymph nodes retrieved on long-term outcome of gastric cancer treatment, 154 patients who had undergone curative resection, with dissection of >15 nodes were retrospectively studied. Dissection of perigastric and extraperigastric lymph nodes, defined as 'extended' (>26 nodes dissected) in 39 cases and 'limited' (< or = 26 nodes dissected) in 115 cases, was performed. A total of 3479 lymph nodes (mean 22.6 per specimen), were dissected and of these 721 showed metastases. A mean of 8.1 lymph node metastases, per metastatic case, was found. Regression analysis showed no independent factor associated with the extent of lymphadenectomy. Depth of wall invasion (p=0.000) and histological growth pattern (p=0.044) were independently associated with the number of lymph nodes involved (pN0, pN1 1-7, pN2 >7). The cumulative 5-year survival rate was 47% in patients without lymph node metastases; 29% in those with 1-7 nodes involved and 17% in those with >8 nodes involved (p=0.002). Receiver operating characteristic (ROC) curve analysis, in 65 nodenegative cancer cases, demonstrated an area under the curve for vital status (alive or dead) of 0.602 (95% CI: 0.473 - 0.721). All node-negative cases with a number equivalent to or exceeding the cutoff point of 23 nodes were alive. ROC analysis showed 11 to be the cutoff number of metastasized lymph nodes in correlation with vital status. Almost all those patients in whom the number of positive nodes was equivalent to, or exceeded the cutoff point had died (area under the ROC curve 0.633; 95% CI: 0.524 - 0.733). ROC analysis showed that the cutoff lymph node ratio, in relation to vital status, was 0.33. The majority of patients at or above this cutoff point had died (area under ROC curve 0.682; 95% CI: 0.574 - 0.776). Multivariate survival analysis showed that lymph node ratio was the only independent prognostic factor (p=0.001). The present findings suggest that, in lymphadenectomy with at least 15 nodes, the number and status of regional nodes dissected, irrespective of the location, provide reliable prognostic information on curatively resected gastric carcinomas. PMID- 15354406 TI - Systemic and haemodynamic toxicity after isolated limb perfusion (ILP) with TNF alpha. AB - The aim of this study was to evaluate the systemic and haemodynamic postoperative effects of ILP with medium-low dose of TNF alpha in patients diagnosed with primary or recurrent limb melanoma or sarcoma, and to compare the resulting toxicity with Systemic Inflammatory Response Syndrome (SIRS). A prospective study on 17 consecutive patients with primary or recurrent limb tumor (melanoma or sarcoma) subjected to ILP with escalating doses of TNF alpha (0.5-2.0mg) was carried out. Seventeen patients with primary or recurrent limb melanoma or sarcoma were subjected to ILP with escalating doses of TNF alpha. ILP was carried out with the standard techniques, blood being warmed at 42 degrees C for an hour. Serial serum TNF alpha determinations were performed before, during and after limb perfusion in nine patients. Systemic and pulmonary haemodynamics, by a radial and pulmonary artery catheter inserted before the induction of anesthesia, were monitored at 5 different times: before the induction of anesthesia (T0), and 6, 12, 24 and 48 hours after treatment (T1-4). Complete isolation of the limb was not always achieved, therefore leakage of TNF alpha occurred frequently during the perfusion in all patients with maximum systemic TNF alpha concentrations ranging from 431 to 111000 pg/ml. After perfusion only two patients showed detectable TNF alpha levels in peripheral blood which returned to baseline values within nine hours. These two patients had serious systemic toxicity: shock and respiratory failure secondary to pulmonary edema. Acute pulmonary edema was also observed in another patient. All three cases required supportive therapy provided by means of mechanical ventilation. In the remaining 14 patients a sepsis-like syndrome was observed. The most significant haemodynamic changes were due to the CO, which rose by 35%, and the SVR, which remained consistently low throughout. A reduction in Hb was observed in all patients (with an average decrease of 4 g/dl), while DO2 and VO2 levels rose, though not to statistically significant levels. Hypoxia occurred in all 14 patients. In three of the remaining 14 cases bilateral pulmonary leaks were noted, however the use of mechanical ventilation was not required. No perioperative death occurred and the aforementioned side effects were all reversible resulting in a patient's mean postoperative ICU permanence of 4 days (range 3 to 7 days). In conclusion, ILP with TNF alpha induces cardiovascular, respiratory and hematological toxicity with haemodynamic parameters being similar to those noted in SIRS probably due to leakage of TNF alpha in the systemic circulation during the perfusion. Nevertheless, this systemic toxicity was short-lived resulting in an acute reaction following a single application. PMID- 15354407 TI - Reduced low-density-lipoprotein cholesterol causing low serum cholesterol levels in gastrointestinal cancer: a case control study. AB - Several epidemiological studies suggested an inverse relation between serum cholesterol level and cancer mortality. We analyzed the relation between gastrointestinal cancers and serum cholesterol levels. A total of 631 patients were recruited as cancer-bearing cases, comprising 181 esophageal cancers, 251 gastric cancers and 199 colorectal cancers. A case-control analysis was conducted on the serum TC, HDL-C, LDL-C and TG levels. TC and LDL-C were significantly lower in cancer-bearers by approximately 15 mg/dl. Furthermore, analyses by cancer site also showed significantly lower TC and LDL-C levels in cancer-bearers than in controls for all three sites. In this analysis, early stage cancer bearers showed a significant decrease in TC levels by approximately 11 mg/dl compared with controls, and also a similar decrease in LDL-C levels. These results suggest that low TC levels are not related to cancer stage. Furthermore, findings of no significant differences in HDL-C and TG between cancer-bearing cases and controls in addition to a specific decrease in LDL-C in cancer-bearers suggest that hypocholesterolemia observed in these cases stems from low LDL-C. However, cancer-bearers and controls showed a similar distribution of TC and LDL C levels. We should be aware that latent cancer bearers may be present among subjects with hypocholesterolemia. PMID- 15354408 TI - Dose-response effect of tomato paste on 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis. AB - We evaluated the dose-response effect of tomato paste on 7,12 dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis using lipid peroxidation, reduced glutathione (GSH) and the GSH dependent enzymes; glutathione peroxidase (GPx), glutathione-S-transferase (GST) and gamma-glutamyltranspeptidase (GGT) as biomarkers of chemoprevention. Hamsters were divided into eight groups of six animals each. The right buccal pouches of animals in group 1 were painted with a 0.5 per cent DMBA in liquid paraffin three times per week. Animals in groups 2 to 4 painted with DMBA as in group 1, received in addition, intragastric administration of tomato paste containing lycopene at concentrations of 2.5, 5 and 10 mgkg(-1)bw, respectively three times per week on days alternate to DMBA application. Groups 5 through 7 were given tomato paste alone. Animals in group 8 served as controls. All animals were killed after an experimental period of 14 weeks. Lipid peroxidation and GSH dependent antioxidants were measured in the buccal pouch, liver and erythrocytes. Diminished lipid peroxidation in the HBP tumours was associated with enhanced levels of GSH and GSH-dependent enzymes. In contrast to the buccal pouch, the liver and erythrocytes of tumour-bearing hamsters exhibited elevated lipid peroxidation accompanied by compromised antioxidant status. Administration of tomato paste significantly reduced the incidence of HBP tumours, modulated lipid peroxidation and enhanced GSH and GSH-dependent enzymes in the pouch, liver and erythrocytes. Among the three doses used, tomato paste containing 5 mgkg(-1)bw lycopene showed the optimum effect. It is suggested that tomato paste exerts its chemopreventive effects by modulating lipid peroxidation and enhancing antioxidants in the target organ as well as in the liver and erythrocytes. PMID- 15354409 TI - Comparative evaluation of cancer chemopreventive efficacy of alpha-tocopherol and quercetin in a murine model. AB - Chemopreventive efficacy and the mode of action of alpha-tocopherol and quercetin, both known to possess anti-oxidant properties, were evaluated during carcinogenesis in uterine cervix of mice by chronic exposure to methylcholanthrene. Oral administration of alpha-tocopherol and quercetin could reduce the genotoxic effect of the carcinogen as revealed by the micronucleus frequency in vaginal epithelial cells. Both could restrict the progression of dysplastic changes in cervix uteri leading to carcinoma in-situ but the influence of the quercetin was more pronounced. A decrease in lipid peroxides following ingestion of these agents suggests that their chemopreventive action is due to their protective role on DNA damage by free radicals. Increased glutathione and glutathione-s-transferase also indicate their involvement in detoxification. While alpha-tocopherol activated the PBL response towards mitogenic stimulation, quercetin had no such effect. PMID- 15354410 TI - Combined effect of arsenic trioxide and sulindac sulfide in A549 human lung cancer cells in vitro. AB - Arsenic trioxide has shown substantial efficacy in treating patients with relapsed or refractory acute promyelocytic leukemia (APL) as well as solid tumors. Arsenic can act through a considerable number of different pathways including mitochondrial respiration and tubulin formation, affecting growth, blood flow, differentiation, and apoptosis. Recent studies on the apoptotic potential of arsenic trioxide have elucidated some of its causal mechanisms, including elevation of intracellular H2O2, inhibition of NF-kappaB activity, and inhibition of GTP-induced polymerization of tubulin. Because of the variety in cellular approaches available to arsenic, it has been hypothesized that the combination of arsenic trioxide and other chemotherapeutic agents may result in cytotoxic synergy. Recent studies have proven this true, with all-trans retinoic acid, IFN-alpha, and ascorbic acid all yielding promising results when used in conjunction with arsenic trioxide. In this study we tested sulindac sulfide, a nonsteroidal anti-inflammatory drug (NSAID) to test its effects with arsenic. Sulindac was used because it functions by some of the same pathways as arsenic, including the mitochondrial apoptotic pathway and the NF-kappaB pathway. Our results show that sulindac sulfide enhances cytotoxicity when combined with arsenic trioxide, and that further studies on the exact mechanisms of their interaction are needed. PMID- 15354411 TI - Strong association of ARK5 with tumor invasion and metastasis. AB - We recently identified a novel human AMPK family member, ARK5, and discovered that is a major factor in Akt-dependent cancer cell survival and migration activity through activation of MT1-MMPs in vitro. The mRNA expression of other AMPK family members and ARK5 was measured using RT-PCR in human colorectal carcinoma cell lines DLD-1, WiDr, HCT-15, SW620, LoVo, SW480, and mRNA expression of AMPK-alpha1, SNARK, MELK and ARK5, but not AMPK-alpha2, was detected in every line. Quantitative-PCR (Q-PCR) to estimate the amount of ARK5 mRNA expression in the cell lines showed that there is a variety of ARK5 expressions among the cell lines and high expression was observed in a cell line derived from the metastatic lesion, LoVo. To determine the effect of ARK5 overexpression on metastasis in vivo, we established human pancreas cancer cell line PANC-1 stably transfected with ARK5 full-length expression vector (P/ARK) and DLD-1 stably transfected with the same vector (D/ARK). Migration assay showed a remarkable increase in the activity both in P/ARK and D/ARK, and an in vivo metastasis assay showed a marked increase of P/ARK in liver metastasis. Based on these observations, it is suggested that ARK5 expression is involved in cancer invasion and metastasis. PMID- 15354412 TI - Tissue evaluation of immune markers in endometrial and cervical carcinomas. AB - Major histocompatibility complex (HLA system) class II molecules including HLA-DR antigens, associate with peptides, which are derived from antigens, for presentation to T4 lymphocytes. Functional and adhesion assays have shown that CD4 molecule interacts with HLA class II molecules, leading to enhanced responses of T4 cells. In the present study, we examined the tissue expression of HLA-DR antigens and the quantitative variance of T4 lymphocytes in a series of 50 "endometrioid" adenocarcinomas of the endometrium and 35 cervical squamous-cell carcinomas. A three-step avidin-biotin immunoperoxidase staining method was applied. As primary antibodies, we used the TAL.1BS monoclonal antihuman HLA-DR alpha (alpha) chain antibody and the OPD4 mouse antihuman antibody; the latter mainly identifies benign T4 lymphocytes. Twenty-four percent (24%) of women with endometrial cancer were high immune responders, while the relative percentage in women with cervical cancer was 40%; the respective tumours were of early clinical and surgical stages. HLA-DR determinants were predominantly expressed in membranes of stromal cells, mainly histiocytes, usually around HLA-DR+ lymphoid cells, as well as on endothelial cells. Greater numbers of OPD4+ aggregated lymphocytes were observed when the tumour stroma was rich in HLA-DR+ cells. Epithelial elements, either cancerous or benign, were seldom HLA-DR+. In those samples, positive immunolabelling was often confined in the intercellular space and did not seem to activate an effective host immune response against neoplastic cells. High expression of HLA-DR molecules in professional antigen presenting stromal cells may be used as a lymphocyte activation marker in endometrial and cervical carcinomas. This activation appears to be an early event in the evolution of invasive endometrial and cervical carcinomas. PMID- 15354413 TI - Expression patterns of cyclins D1, E in laryngeal epithelial lesions: correlation with other cell cycle regulators (p53, pRb, Ki-67 and PCNA) and clinicopathological features. AB - The expression of cell-cycle progression molecules cyclin D1 and cyclin E were immunohistochemically examined in a series of 64 squamous cell invasive carcinomas of the larynx, 10 in situ carcinomas, 34 cases of dysplasia, 11 papillomas and 23 cases of keratosis. The results of their expression were compared with two cell-cycle implicated tumor suppressor proteins p53 and pRb as well as with two proliferation associated indices PCNA and Ki-67 in an attempt to elucidate their potential role in the pathogenesis and progression of these lesions. Nuclear staining for cyclin D1 and E (>5% positive cells) was observed in 19% and 39.7% of the laryngeal carcinomas, respectively. Significantly elevated levels of cyclin D1 and E in invasive laryngeal carcinomas compared with in situ carcinomas were revealed (p=0.045 and p=0.0003, respectively). High levels of cyclin D1 and E expression were correlated with increased Ki-67 score (p=0.037 and 0.017 respectively). A significant positive correlation between cyclin D1 and E was also detected in carcinomas (p=0.018). Decreased levels of cyclins D1 and E in the group of in situ carcinomas compared with those of dysplastic cases and papillomas were also observed. In the dysplastic lesions cyclin D1 expression was correlated with pRb expression (p=0.02). In the cases of keratosis cyclins D1 and E expression were correlated with pRb (p=0.002 and p=0.036, respectively), while cyclin D1 was associated with PCNA (p=0.008) and Ki 67 score (p=0.009). The prognostic significance of cyclins D1, E in determining the risk of recurrence and overall survival with both univariate (long-rang test) and multivariate (Cox regression) methods of analysis showed no statistically significant differences. We conclude that the expression of cyclins D1 and E in squamous cell carcinomas of the larynx does not seem to have a prognostic significance. In addition, their expression may be involved in the development of laryngeal lesions, implicated in cell proliferation, with other cell cycle related proteins, probably by different molecular pathways. PMID- 15354414 TI - An association of polymorphism of DNA repair genes XRCC1 and XRCC3 with colorectal cancer. AB - Variability in DNA repair genes may contribute to human cancer risk. We performed a case-control study (51 cases and 100 controls) to test the association between two polymorphisms: Arg399Gln in the XRCC1 gene and Thr241Met in the XRCC3 gene and colorectal cancer risk. Genotypes were determined in tumour tissue and distant mucosa samples by PCR RFLP with the NciI restriction enzyme for XRCC1 and NcoI for XRCC3. Cancer occurrence was strongly associated with the XRCC3 Met/Met polymorphic variant (OR = 9.45; (95% CI 8.77-11.65)), whereas Thr/Thr and Thr/Met variants were associated with significant reduction in colorectal cancer risk (OR = 0.16; 95% CI 0-0.26 and OR = 0.26; 95% CI 0.25-0.27, respectively). Weak association was found between the XRCC1 Arg/Arg and Gln/Gln variants and the risk of colorectal cancer (OR = 1.28; 95% CI 1.00-1.84 and OR = 1.13; 95% CI 0.85 2.34, respectively). Gene-gene interaction between the XRCC1 Arg/Arg and XRCC3 Met/Met homozygous variants slightly increased the risk (OR = 10.50; 95% CI 5.67 14.79). Both polymorphisms were not associated with colorectal cancer progression. PMID- 15354415 TI - Cytogenetic and fluorescence in situ hybridization monitoring in Ph+ Chronic Myeloid Leukemia patients treated with imatinib mesylate. AB - Imatinib mesylate determines a favorable clinical course in most Ph positive Chronic Myeloid Leukemia (CML) patients in the chronic phase. Cytogenetic response is usually evaluated by analyzing 20-25 bone marrow metaphases using standard banding techniques. Since this methodology has very low sensitivity, we compared the results obtained by standard banding techniques to the ones obtained by fluorescent in situ hybridization (FISH). This was also done to identify any possible discrepancies between the two techniques. We analyzed 40 Ph+ CML patients in the chronic phase who had previously been treated with interferon alpha (IFNalpha) and who were receiving imatinib. The studies were performed by utilizing the same BM cell samples fixed in acetic acid/methanol, before imatinib therapy and then quarterly. Comparison of cytogenetic results to FISH results at 3 and 6 months of imatinib treatment showed that some patients who had achieved major cytogenetic response (i.e.<35% of examined metaphases showing Ph), showed retention of a higher number of persisting Ph+ cells when examined by FISH, and they did not achieve major FISH response (i.e. <35% of examined interphase cells show the BCR-ABL fusion signal). The discrepancy we found between the results that were obtained by analyzing metaphases and interphase cells disappeared in the subsequent examinations. Moreover, we found that 4 patients (10%) were still Ph+ in all the metaphases we examined even though they achieved excellent clinical response. On the basis of this small series of patients, we suggest that cytogenetic evaluation of patients on imatinib therapy should be performed by utilizing the classic banding technique (metaphase examination), but also by using the FISH technique (interphase examination), since the two methodologies may provide different results. PMID- 15354416 TI - A selective cyclooxygenase-2 inhibitor, NS398, inhibits cell growth and induces cell cycle arrest in the G2/M phase in human esophageal squamous cell carcinoma cells. AB - It is well known that cyclooxygenase (COX) -2 is expressed in a variety of human malignant solid tumors, associated with tumor angiogenesis, cell proliferation and inhibition of apoptosis. Here, we examined the effect of NS398, a selective COX-2 inhibitor, on two human esophageal squamous cell carcinoma (SCC) cell lines, TE-1 and TE-12. Western blot analysis confirmed the expression of COX-2 in TE-12, but not in TE-1. Treatment with 100microM NS398 suppressed the cell viability in TE-12 (48.6% of control) after 48 hours, in contrast to showing no effects in TE-1. The apoptotic index was extremely low in both cell lines after the treatment. NS398 clearly increased the number of cells in the G2/M phase and decreased the cells in the G1 and S phases in TE-12, but not TE-1. A pre-G1 fraction was not noted in either cell line. Moreover, TE-12 cells showed a decrease in the expression levels of cyclin B1 and an increase in p27Kip1. These findings suggest that NS398 inhibits cell growth and induces G2/M arrest in human SCC cells expressing COX-2. PMID- 15354417 TI - Effect of all-trans retinoic acid on telomerase activity in ovarian cancer cells. AB - Vitamin A is an essential nutrient important for growth, vision, embryonic development, immune response and reproduction. Various retinoids have been shown to be effective chemotherapeutic and chemopreventive agents for a number of human cancers. Telomeres are nucleoprotein structures found at the end of chromosomes. During cellular division, the telomeres in normal cells shorten progressively and thus, function as a "molecular clock". Telomerase is a ribonucleoprotein complex that extends and maintains telomeres. Activation of telomerase is required for cells to overcome proliferative crisis. Telomerase activation is observed in 90% of human cancers, but not in normal somatic cells. We examined the role of telomerase in mediating the growth suppression of ovarian carcinoma cells by all trans-retinoic acid (ATRA). Using a number of cell lines with varying levels of growth sensitivity to ATRA, we found that cells that exhibit ATRA-dependant suppression of growth also contained significantly reduced telomerase activity. We also observed a reduction in expression of the telomerase components, hTERT and hTR in ATRA treated ovarian carcinoma cells. Our results suggest that one mechanism by which ATRA acid inhibits cancer cell growth is by suppressing telomerase activity, thereby pushing cells to proliferative crisis. PMID- 15354418 TI - Repression of MLH1 and MGMT genes in colon mucosa adjacent to implanted cancer in athymic mouse. AB - Hyperplastic mucosa adjacent to colon cancer, being a reactive change, accelerates cancer progression and its metastasis through expression of angiogenic factors. We investigated promoter methylation in hyperplastic mucosa adjacent to orthotopic KM12SM colon cancer in mice. In the hyperplastic mucosa adjacent to KM12SM tumors in the cecum of athymic mice, reductions in the levels of the mutL homologue 1 (MLH1) and O6-methylguanine-DNA methyltransferase (MGMT) proteins were detected by immunohistochemistry and immunoblotting. To examine the effects of growth factors and cytokines on promoter methylation and repressed expression of the MLH1 and MGMT genes, a rat intestinal epithelial cell line, IEC6, was treated with epidermal growth factor (EGF) and interleukin (IL)-15 for 35 days. Protein levels of MLH1 and MGMT were reduced in EGF- and IL-15-treated IEC6 cells. A methylation-sensitive restriction enzyme assay revealed that CpG methylation was present in the promoter regions of the MLH1 and MGMT genes in DNAs extracted from hyperplastic mucosa adjacent to KM12SM tumors. These findings suggest that promoter CpG methylation affects expression of MLH1 MGMT genes in hyperplastic mucosa adjacent to colon cancer. PMID- 15354419 TI - CSE1/CAS overexpression inhibits the tumorigenicity of HT-29 colon cancer cells. AB - We previously reported that CSE1/CAS (CAS) overexpression in HT-29 human colon cancer cells enhances the formation of the E-cadherin/beta-catenin complex, stimulates intercellular junction formation, and stimulates polarization of HT-29 cells. Since both E-cadherin/beta-catenin interaction and epithelial cell polarization are critically related to the tumorigenicity of carcinoma cells, we studied the role of CAS in the tumorigenicity of HT-29 colon carcinoma cells. CAS overexpression in HT-29 cells decreased the intercellular gaps and increased the compactness of cell colonies. Our results show that CAS expression inhibited migration and growth of HT-29 cancer cells. In the soft agar anchorage independent growth assays, CAS overexpression inhibited the colony size of HT-29 cells by 74%, and inhibited colony formation number of HT-29 cells by 38%. CAS overexpression also inhibited the growth of HT-29 cells in nude mice. Our results indicate that CAS inhibits the tumorigenicity of HT-29 human colon cancer cells and, thus, it is worthwhile to further study CAS's possible role in the control of human colon cancer. PMID- 15354420 TI - Synergistic anti-tumor effect of ultrasound and hematoporphyrin on sarcoma180 cells with special reference to the changes of morphology and cytochrome oxidase activity of tumor cells. AB - This study is aimed at evaluating the inhibitory effects of the association of hematoporphyrin and ultrasound at variable intensities with a fixed frequency of 1.1MHz in tumor nodules. Specifically, the effects were studied both in solid and ascitic S180 tumors transplanted in mice by clinical, cytochemical and ultrastructural evaluation. The results indicated that the use of hematoporphyrin alone had no significant effect on destroying tumor cells. The ultrasound alone had little effect. Interestingly, the inhibition was much more effective when hematoporphyrin was combined with ultrasound. The inhibition was 3 times better than ultrasound alone and 8 times better than hematoporphyrin used alone. Our results also indicated that the changes on cell structure and cytochrome oxidation activity are important factors that could inhibit tumor cell growth and induce cell death. Apoptosis of tumor cells could be induced by hematoporphyrin. Our study investigated the killing mechanism on S180 tumor cells by using hematoporphyrin and low frequency ultrasound at cell, tissue and individual level. PMID- 15354421 TI - Myasthenic crisis following cisplatin chemotherapy in a patient with malignant thymoma. AB - Malignant thymomas are associated with numerous autoimmune disorders including myasthenia gravis. Myastenia gravis is characterized by antibodies against the acetylcholine receptors located on the neuromuscular junction of the skeletal muscle. We present a case with malignant thymoma who developed myasthenia crisis while he was treated with cisplatin chemotherapy. PMID- 15354422 TI - Angiotropic large cell lymphoma. AB - A 67 year old man developed "status epilepticus" and died with acute respiratory failure. An autopsy and concomitant histological examination, revealed intravascular infiltrate of lymphoid cells throughout the body. Immunohistochemical studies showed intense positive staining for leukocyte common antigen (LCA), L26 and CD20 and negative staining for Factor VIII- related antigen, and CD5, demonstrated that the neoplastic cells were of B-cell lineage. PMID- 15354423 TI - Natural history of a patient with malignant nonfunctioning islet cell tumor associated with unresectable multiple liver metastases. AB - Islet cell tumors of the pancreas are uncommon. Approximately 15% of islet cell tumors are nonfunctioning and have a higher malignancy rate than their functioning counterparts. Though, because of the rarity of malignant nonfunctioning islet cell tumors, the natural history of a patient with this tumor has not been clearly defined. We describe a young patient with unresectable malignant nonfunctioning islet cell tumor associated with multiple liver metastases. He was treated with palliative therapies to improve his quality of life, but did not undergo surgical removal of tumors or systemic chemotherapy. He survived for 46 months since laparotomy for histological diagnosis. Our findings may represent the natural history of patients with unresectable malignant nonfunctioning islet cell tumor, and suggest that palliative therapy may contribute not only to the improvement of a patients' quality of life but also the prolongation of survival. PMID- 15354424 TI - Recent experimental data on Extremely Low Frequency (ELF) magnetic field carcinogenic risk: open questions. AB - The NTP experimental studies on Extremely Low Frequency (ELF) magnetic fields (two-year studies in rats and mice and magnetic field promotion/DMBA initiation in female rats studies, NTP 1999) are of main interest because of the value and completeness of the information produced on neoplastic and non-neoplastic effects. Ajoint evaluation of the two-year study data indicates a thyroid C-cell focal hyperplasia increase in female rats at the same exposures at which the neoplasm increment of the same cells has been observed only in male rats (on which the "equivocal evidence" NTP classification is based). The significant exposure-related trichoepithelioma and trichoepithelioma plus other skin neoplasms reported for male rats in the two year studies finds some support in the female rat data relative to trichoepithelioma reported in incidence summary of the NTP 26-week initiation/promotion study, even if the experimental designs of the two studies are different. An indication of exposure-related increase of single mammary carcinomas emerges from the summary of neoplasm incidence, together with a dose-related decrease of mammary gland multiple carcinomas. A significant decrease of lung alveolar/bronchiolar adenoma incidence was observed in exposed groups in both mouse genders. It seems improbable to attribute the above effects only to chance. The present debate, dividing science in two opposite fields: one of them denying any appreciable carcinogenic effect of magnetic fields, and the other one hypothesising their dramatic effects, are presently devoid of reliable and exhaustive scientific support, which could only be provided by further research. PMID- 15354425 TI - Progressive ascending paraparesia in an acute lymphoblastic leukemia patient: masked but not fallacious. PMID- 15354426 TI - Development and application of polymeric monolithic stationary phases for capillary electrochromatography. AB - Monolithic columns for capillary electrochromatography are receiving quite remarkable attention. This review summarizes results excerpted from numerous papers concerning this rapidly growing area with a focus on monoliths prepared from synthetic polymers. Both the simplicity of the in situ preparation and the large number of readily available chemistries make the monolithic separation media a vital alternative to capillary columns packed with particulate materials. Therefore, they are now a well-established stationary phase format in the field of capillary electrochromatography. A wide variety of synthetic approaches as well as materials used for the preparation of the monolithic stationary phases are presented in detail. The analytical potential of these columns is demonstrated with separations involving various families of compounds and different chromatographic modes. PMID- 15354427 TI - Sol-gel stationary phases for capillary electrochromatography. AB - A review is presented on the current state of the art and future trends in the development of sol-gel stationary phases for capillary electrochromatography (CEC). The design and synthesis of stationary phases with prescribed chromatographic and surface charge properties represent challenging tasks in contemporary CEC research. Further developments in CEC as a high-efficiency liquid-phase separation technique will greatly depend on new breakthroughs in the area of stationary phase development. The requirements imposed on CEC stationary phase performance are significantly more demanding compared with those for HPLC. The design of CEC stationary phase must take into consideration the structural characteristics that will provide not only the selective solute/stationary phase interactions leading to chromatographic separations but also the surface charge properties that determine the magnitude and direction of the electroosmotic flow responsible for the mobile phase movement through the CEC column. Therefore, the stationary phase technology in CEC presents a more complex problem than in conventional chromatographic techniques. Different approaches to stationary phase development have been reported in contemporary CEC literature. The sol-gel approach represents a promising direction in this important research. It is applicable to the preparation of CEC stationary phases in different formats: surface coatings, micro/submicro particles, and monolithic beds. Besides, in the sol-gel approach, appropriate sol-gel precursors and other building blocks can be selected to create a stationary phase with desired structural and surface properties. One remarkable advantage of the sol-gel approach is the mild thermal conditions under which the stationary phase synthesis can be carried out (typically at room temperature). It also provides an effective pathway to integrating the advantageous properties of organic and inorganic material systems, and thereby enhancing and fine-tuning chromatographic selectivity of the created hybrid organic-inorganic stationary phases. This review focuses on recent developments in the design, synthesis, characterization, properties, and applications of sol-gel stationary phases in CEC. PMID- 15354428 TI - Approaches to imprinted stationary phases for affinity capillary electrochromatography. AB - The combination of two highly attractive technologies in analytical chemistry, namely molecular imprinting and capillary electrochromatography, is reviewed. Molecular imprinting represents an approach to incorporate affinity binding sites for one or a class of target molecules in network polymers whereas capillary electrochromatography is a technique that profits from the high separation efficiency of electrodriven separations and the high selectivity and capacity available with liquid chromatography. The review discusses and compares the various approaches taken, to combine these concepts aiming at robust, reproducible and easily available capillary-based affinity separation media. PMID- 15354429 TI - Recent highlights in stationary phase design for open-tubular capillary electrochromatography. AB - This review examines the most recent innovations made to achieve high performance in open-tubular capillary electrochromatography (OT-CEC) separations, focusing on the ingenious chemical and physical solutions made to increase the surface area and equip the stationary phase with exploitable selectivity. Among the approaches taken are chemically bonded ligands, etching with chemical bonding, sol-gels, molecularly imprinted polymers, porous layers, physically attached or adsorbed phases, and nanoparticle coatings. Particularly noteworthy are modern developments with macrocyclic receptor ligands, nanoparticles and open channel electrochromatography on-chip. PMID- 15354430 TI - Planar electrochromatography. AB - Recent developments in planar electrochromatography (PEC) in both the normal phase and the reversed-phase modes, and at both atmospheric and elevated pressure, are reviewed. Other forced-flow techniques in planar chromatography are also briefly covered. Mobile phase migration in PEC is primarily due to electroosmotic flow, which is controlled by the applied electric field. Capillary mediated flow is an important secondary contributor to migration, and occurs because the layer is unsaturated as a consequence of liquid evaporating from the layer due to Joule heating. The magnitude of the electric field and the concentration of ions in solution are important variables that control both electroosmotic flow and Joule heating. Separations are faster and more efficient than those obtained by conventional planar chromatography, provided appropriate experimental conditions are selected. With inappropriate conditions, either mobile phase accumulates on the surface of the sorbent layer, or Joule heating causes excessive evaporation. The former results in poor spot shape, and the latter can cause the layer to dry. Good separations are obtained when there is a balance between these two effects. The problems associated with mobile phase accumulating on the surface of the sorbent layer, and with excessive evaporation of mobile phase, do not occur with pressurized planar electrochromatography. This technique is performed at high pressure, under conditions that allow heat to be removed form the sorbent layer. This allows the use of a substantially higher electric field than in PEC, and results in a high mobile phase flow rate. PMID- 15354431 TI - Chip electrochromatography. AB - Electrochromatography (EC) in microfluidic chips is emerging as an attractive alternative to capillary electrophoresis (CE) for on-chip separations. This review summarizes recent developments in the rapidly growing area of chip electrochromatography with a focus on "column" technologies. Relevant achievements are summarized according to the types of stationary phase used for the separations including open channels, microfabricated structures, and channels packed with beads or containing a porous monolith. The advantages and disadvantages of each, as well as practical aspects of their application, are discussed. The analytical performance of these devices is demonstrated with separations involving various families of compounds mostly in the reversed-phase chromatographic mode. PMID- 15354432 TI - Capillary electrochromatography of peptides and proteins. AB - This review surveys the accomplishments in the separation of peptides and proteins by capillary electrochromatography (CEC) over the last decade. A significant number of research articles have been published on this topic since the last review. Peptide and proteins separations have been carried out in all three formats of CEC, i.e., packed bed, continuous bed and open-tubular (OT) format. In addition to electrophoresis, different chromatographic modes have been successfully exploited with the most prevalent being reversed-phase mode followed by ion-exchange. Although many researchers continue to use model proteins and peptides primarily to evaluate the performance of novel stationary phases some researchers have also applied CEC to the analysis of real-life samples. The potential of CEC to yield complementary information and sometimes a superior separation with respect to established techniques, i.e., microbore HPLC and capillary electrophoresis has been demonstrated. Instrumental modifications in order to facilitate coupling of CEC to mass spectrometry have further upgraded the value of CEC for proteomic analysis. Capillaries are still the separation vehicle of choice for most researchers yet the microfluidic platform is gaining momentum, propelled particularly by its potential for multitasking, e.g., performing different chromatographic modes in series. PMID- 15354433 TI - Review coupling of capillary electrochromatography to mass spectrometry. AB - This review discusses the development of capillary electrochromatography (CEC) coupled to mass spectrometric (MS) detection over the last few years. Major topics addressed are instrumental setups employed and applications of this technology published in the recent literature. The instrumental section includes a discussion of the most commonly used interfaces for the hyphenation of CEC and MS as well as ionization techniques. Applications reviewed in this paper come from a variety of different fields such as the analysis of biomolecules like proteins, peptides, amino acids or carbohydrates, chiral separations or the analysis of pharmaceutical an their metabolites in a series of matrices. PMID- 15354434 TI - Capillary electrochromatography in the size-exclusion mode. AB - In this review the literature on the application of capillary electrochromatography (CEC) for size-based separations of macromolecules is summarized. Opportunities and limitations of CEC specially related to the size exclusion mode (SEEC) are indicated. Applications with synthetic polymer samples as well as with biomacromolecules (polysaccharides, proteins) are shown. The prospects for a further development and application of SEEC are discussed. PMID- 15354435 TI - Peak compression effects in capillary electrochromatography. AB - Peak compression in CEC is a phenomenon that can generate very narrow peaks with extremely high efficiencies that defy current chromatographic theory. This review article summarises the content of publications in this area up to this date. Two main types of peak compression effects have been observed in the literature. First, an irreproducible and hard to control focusing effect of unclear origin, observed on strong cation exchangers. Second, a reproducible continuous stacking effect caused by sample composition induced system zones demonstrated on several types of stationary phases. PMID- 15354436 TI - Developments in the use of soluble ionic polymers as pseudo-stationary phases for electrokinetic chromatography and stationary phases for electrochromatography. AB - This article reviews the development, characterization and application of soluble ionic polymeric materials as pseudo-stationary phases for electrokinetic chromatography and as stationary phases for electrochromatography since 1997. Polymeric pseudo-stationary phases for electrokinetic chromatography, including cationic polymers, anionic siloxane and acrylamide polymers, polymerized surfactants (micelle polymers), and chiral polymers are reviewed. Also reviewed are suspended molecularly imprinted polymer micro-particles. Application of polymeric pseudo-stationary phases with electrospray ionization mass spectrometric detection is presented. Recent progress in the development and characterization of physically adsorbed stationary phases for electrochromatography using polymers of the same or similar chemistry is also reviewed. PMID- 15354437 TI - Affinity chromatography with monolithic capillary columns I. Polymethacrylate monoliths with immobilized mannan for the separation of mannose-binding proteins by capillary electrochromatography and nano-scale liquid chromatography. AB - Monolithic capillary columns with surface-immobilized mannan have been introduced for affinity-based micro-column separations by nano-liquid chromatography (nano LC) and capillary electrochromatography (CEC). Two kinds of polymethacrylate monoliths were prepared, namely poly(glycidyl methacrylate-co-ethylene dimethacrylate) and poly(glycidyl methacrylate-co-ethylene dimethacrylate-co-[2 (methacryloyloxy)ethyl]trimethyl ammonium chloride) to yield neutral and cationic macroporous polymer, respectively. While neutral monoliths with immobilized mannan were only useful for affinity nano-LC, the cationic monoliths with surface bound mannan were useful in both affinity nano-LC and affinity CEC. The cationic monoliths allowed a relatively high electro-osmotic flow (EOF) when mannan was immobilized to the epoxy monolith via a positively charged spacer arm, triethylenetetramine. The neutral monoliths exhibited lower permeability under pressure-driven flow (PDF) than the cationic monoliths indicating that the latter had wider flow-through pores than the former. Both types of monoliths with immobilized mannan exhibited strong affinity toward mannose-binding proteins (MBP) such as the plant lectins concanavalin A and Lens culinaris agglutinin and a mammalian lectin (e.g. rabbit serum mannose-binding protein). Due to the strong binding affinity, the monoliths with surface bound mannan allowed the injection of large volume of rabbit serum and to isolate in a single run the mannose binding protein in an amount sufficient to run with it sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), thus demonstrating their capability in "nano-proteomics". PMID- 15354438 TI - Development of reactive thiol-modified monolithic capillaries and in-column surface functionalization by radical addition of a chromatographic ligand for capillary electrochromatography. AB - Reactive thiol-modified capillary columns for capillary electrochromatography (CEC) were developed by transforming the pendent 2,3-epoxypropyl groups of poly(glycidyl methacrylate-co-ethylene dimethacrylate) (poly(GMA-co-EDMA)) monoliths into 3-mercapto-2-hydroxy-propyl residues by a nucleophilic substitution reaction, employing sodium-hydrogen sulfide as nucleophilic reagent. Conditions for this modification reaction were systematically optimized with respect to different parameters, such as reaction temperature, pH-value, reaction time, type and concentration of organic modifier, and concentration of the sodium hydrogen sulfide solution. The amount of thiol groups that was generated on the monolith surface was determined directly in the capillaries by a disulfide exchange reaction employing 2,2'-dipyridyl disulfide (DPDS). This reaction in the capillary liberates pyridine-2-thione in equimolar amount to the surface sulfhydryls, which was collected into a vial and determined photometrically at 343 nm by RP-HPLC. About 17% of the total lateral epoxide moieties of the monolithic substrate could be transformed to reactive sulfhydryl groups, which corresponds to about 0.7 mmol g(-1) monolithic polymer, with a column-to-column repeatability of 3.2% R.S.D. The reactive thiol groups can be utilized to attach any chromatographic ligand with appropriate anchor in a second step, e.g. by radical addition, graft polymerization, nucleophilic substitution, disulfide formation or Michael addition reaction. To demonstrate the feasibility of the concept, we chose an anion exchange type chromatographic ligand based on a quinine derivative, O-9-tert-butylcarbamoylquinine (t-BuCQN) which was attached to the monolith in a radical addition reaction, for a further in-column surface functionalisation. About 78% of the sulfhydryl groups were derivatized with t BuCQN as determined from differential DPDS assays before and after the selector immobilization reaction. The applicability of these surface-functionalised monolithic capillary columns could be shown by an electrochromatographic separation of the enantiomers of N-3,5-dinitrobenzoyl-leucine, which performed fairly well compared to an analogous capillary that was fabricated by an in situ copolymerization approach. PMID- 15354439 TI - Comparison of a three-peptide separation by capillary electrochromatography, voltage-assisted liquid chromatography and nano-high-performance liquid chromatography. AB - A mixture of three peptides was separated by capillary electrochromatography (CEC), nano-HPLC and voltage-assisted LC. In the latter case the charged analytes migrate through a neutral stationary phase driven by electrophoresis while their interaction with the stationary phase provides the basis for a chromatographic separation. The stationary phases used were poly(glycidyl methacrylate-co ethylene dimethacrylate)-based monoliths that could be used directly as neutral "C1"-type columns for voltage-assisted LC and nano-HPLC, while their application in CEC became possible after derivatization of the epoxy groups with ionogenic N ethylbutylamine functions. The separation of the peptide mixture was possible in all three modes. Highest plate numbers and resolutions were obtained under voltage-assisted conditions. The elution order showed dependencies on the charge density but also on the hydrophobicity of the peptides and was different in the three investigated chromatographic modes. The effect of changes in the ionic strength and the organic solvent content of the mobile phase on the resolution and the migration behavior of the peptides was investigated and showed the expected behavior. Voltage-assisted LC is suggested as an alternative to CEC for the separation of charged analytes by electrochromatography. PMID- 15354440 TI - Capillary electrochromatography with polymeric continuous beds synthesized via free radical polymerization in aqueous media using derivatized cyclodextrins as solubilizing agents. AB - A novel synthetic route to amphiphilic acrylamide-based monolithic stationary phases for capillary electrochromatography (CEC) employing water-soluble cyclodextrins as solubilizing agents was explored. N,N'-Octamethylenebisacryamide and N,N'-dodecamethylenebisacryamide were synthesized and their solubilization in aqueous solution with derivatized and underivatized cyclodextrins of different cavity size was studied. Amphiphilic stationary phases were synthesized by free radical copolymerization of the bisacrylamide-cyclodextrin host-guest complexes with hydrophilic monomers and an additional hydrophilic cross-linker in aqueous solution. Complex formation in solution and removal of the complexed cyclodextrin from the polymer during synthesis was studied with 1H-NMR and solid state 13C-NMR spectroscopy and cyclodextrin-modified micellar electrokinetic chromatography. The impact of the incorporated alkylene groups in the acrylamide-based macroporous polymer on retention was studied with neutral solutes by CEC in the normal-phase elution mode and in the reversed-phase elution mode. Batch-to-batch reproducibility of the synthesis procedure and day-to-day repeatability of the separations achieved were investigated. With these capillaries, a sufficiently high electroosmotic flow velocity, a high reproducibility and repeatability of separation parameters and high plate numbers (up to 200,000 m(-1) were obtained. PMID- 15354441 TI - Homogeneous reversed-phase agarose thermogels for electrochromatography. AB - A method for the derivatization of agarose by covalent attachment of hydrophobic ligands for reversed-phase (RP) chromatographic separation and ionic groups for generation of electroosmosis under electrochromatographic conditions in the capillaries or microfluidic channels filled with the thermogel of this agarose derivative is described. The product renders a capability of reversible thermogelation. The thermogels formed provide sufficient hydrophobicity and electroosmosis for the separations of the analytes under RP mobile-phase conditions and electric field applied. The gels may be used repeatedly without loss of resolution. They are thermally replaceable and UV transparent (providing possibility in column/in-gel detection), require no covalent attachment to the capillary inner wall (or microchip channel), and are suitable for isocratic or gradient operation in the aqueous-organic mobile phases. PMID- 15354442 TI - Polystyrene monolithic column functionalized with copper-iminodiacetate complex as a stationary phase for open tubular capillary electrochromatography. AB - Polystyrene-divinylbenzene copolymer functionalized with copper-iminodiacetate complex was prepared in situ for the capillary electrochromatographic separation of amino acids and oligopeptides. The steps included silanization of the fused silica capillary column, functionalization, polymerization, hydrolysis and complexation. For having a homogeneous polymerization system in the selected porogen, the functional monomer was obtained from the reaction of 4-vinylbenzyl chloride and diethyl iminodiacetate. Scanning electron microscopy showed that the polymeric monolith column was an open tubular column with a thickness of 0.3 microm. Various modes of hydrolysis were investigated via the electroosmatic flow measurement to find the optimization condition prior to introducing the copper ion. The influence of pH, composition and concentration of mobile phase as well as the organic modifier were investigated. Judging from our results, the recognition performance of the prepared stationary phase might be integrated from the mechanisms of ligand exchange, electrophoretic mobility, hydrophobic interaction, and hydrogen bonding. PMID- 15354443 TI - Monolithic columns with mixed modes of reversed-phase and anion-exchange stationary phase for capillary electrochromatography. AB - A capillary electrochromatography (CEC) monolithic column with mixed modes of reversed-phase and anion-exchange stationary phases was prepared by in situ polymerization of 2-(methacryloxy)ethyltrimethylammonium methyl sulfate (MEAMS) and ethylene dimethacrylate (EDMA) in a binary porogenic solvent consisting 1 propanol and 1,4-butanediol. The ammonium groups on the surface of the stationary phase generate an electroosmotic flow (EOF) from cathode to anode, and serve as a strong anion-exchange stationary phase at the same time. The EOF of the stationary phase can be determined by the amount of MEAMS monomer in reaction mixtures during the polymerization. The monolithic stationary phases exhibited reversed-phase chromatographic behavior toward neutral solutes. For charged solutes, hydrophobic as well as electrostatic interaction/repulsion with the monoliths was observed. Separations of aromatic compounds and basic compounds on the prepared column were performed under the mode of CEC. Peak tailing of basic compounds was avoided and the efficient separation of aromatic acids was achieved using neutral mobile phase due to the same direction of EOF and electrophorestic mobility of negatively charged solutes. PMID- 15354444 TI - Polymerised bicontinuous microemulsions as stationary phases for capillary electrochromatography. Effect of pore size on chromatographic performance. AB - Monolithic columns for capillary electrochromatography (CEC) were prepared by in situ polymerisation of bicontinuous microemulsions containing butyl methacrylate. The resulting monoliths were found to be permeable to mobile phase flow and their behaviour as CEC stationary phases was investigated. It was found that the monoliths were able to separate a simple test mixture of phthalates, but that efficiencies were low. However, several advantages of the monoliths compared to conventional ODS packed columns were found: preparation time is short, many columns can be prepared from the same batch of microemulsion and column conditioning is much faster. The columns show promise as stationary phases for CEC, but more development is required to improve efficiencies. PMID- 15354445 TI - Towards a miniaturised system for dynamic field gradient focused separation of proteins. AB - Separation and focusing of proteins is described in a miniaturised dynamic field gradient focusing device with a 2.5 cm x 0.1 cm channel filled with a porous polymer monolith. The separation channel is in contact with a parallel electric field channel with five individually addressable electrodes through a porous glass membrane so that a variable field can be generated that drives charged proteins electroosmotically against a constant hydrodynamic flow. Separated pre stained proteins were detected by means of a digital camera and background subtraction. PMID- 15354446 TI - Sample injection in capillary electrochromatography by heart-cut technique. AB - The splitter working in heart-cut regime was used for sample injection in capillary electrochromatography. The principle was implemented in an automated microgradient system allowing to inject from microlitre down to nanolitre volumes with high repeatability and minimal extra-column band broadening. The apparatus is able to deliver discrete volumes of liquids at a preset volumetric flow rate and to stop and restore the flow at any moment. This brings a high degree of liquid manipulation flexibility. An extremely low split ratio is sufficient during the analysis, which saves mobile phase consumption substantially. The key parameters influencing the function of the heart-cut splitter were characterised. The function of the apparatus was demonstrated under isocratic, preconcentration and gradient capillary electrochromatography separation conditions. In all cases the statistic evaluation of the main parameters was performed, showing that high repeatability of retention times, peak heights and areas was achieved. PMID- 15354447 TI - Silica sol-gel/organic hybrid material for protein encapsulated column of capillary electrochromatography. AB - A new-type of sol-gel/organic hybrid composite material using gelatin or chitosan with tetramethoxysilane was developed for the bovine serum albumin (BSA) encapsulated monolithic column for capillary electrochromatography (CEC). The composite monolith was used to immobilize BSA in a fused-silica capillary. The addition of gelatin and chitosan to the alkoxysilane enabled the enantioseparation of Trp. A very small amount of these polymers were effective for the enantioseparation. Especially, the monolithic column prepared from chitosan with tetramethoxysilane showed a high enantioselectivity for Trp enantiomers and the value (alpha' = t2/t1, t1: fast eluted enantiomer, t2: second eluted enantiomer) reached 1.15 on CEC mode. Furthermore, the composite materials exhibited a higher stability compared to the silica sol-gel column. These results showed that the sol-gel/organic hybrid composite was useful as a monolithic matrix for the BSA-encapsulated column for CEC. PMID- 15354448 TI - On-column derivatization-capillary electrochromatography with o phthalaldehyde/alkylthiol for assay of biogenic amines. AB - The elution behaviors of the biogenic amines, histamine (HA) and its metabolite methyl histamine (MHA), were evaluated by means of on-column derivatization (OCD) capillary electrochromatography (CEC) which employed a monolithic octadecylsilica (ODS) capillary column (20 cm of effective length x 50 microm of inner diameter). Five kinds of alkylthiols, e.g., 2- hydroxyethylthiol (or 2-mercaptoethanol (2 ME)), ethanethiol (ET), 1-propanethiol (1-PT), 2-methyl-1-propanethiol (2-MPT) and 1-butanethiol (1-BT) were separately presented at 5 mM each in the OCD-CEC separation run buffer consisting of 60% acetonitrile in 5 mM o-phthalaldehyde (OPA)-10 mM borate buffer (pH 10). When 2-ME was present in the run buffer solution, both derivatives corresponding to HA and MHA migrated separately, but closely together through the capillary column. Replacement of 2-ME with 1-BT in the run buffer solution caused a delay in their elution profiles on the electrochromatogram and the separation between those two peaks became remarkably improved. The elution times of HA and MHA followed the increase in alkyl chain length or hydrophobicity of thiol, 1-BT > 2-MPT > 1-PT > ET > 2-ME. Performance of on-line preconcentrations of HA and MHA was also evaluated by varying the electrokinetic injection voltage from 1 kV to 8 kV. The peak area counts corresponding to HA recorded about 50 times higher when 2 kV was applied for 240 s to a 0.1 mM HA solution than when 8 kV was applied for 5 s. This method was next applied to a sample of human urine spiked with HA and MHA at levels of 0.1 microM each. Although HA and MHA peaks were not identifiable among the peaks corresponding to the materials in the urine matrix when OPA/2-ME was employed in a run buffer for the OCD-CEC, the separation and identification of their peaks became possible by replacing 2-ME with 1-BT in the run buffer solution. PMID- 15354449 TI - On-chip electrochromatography using sol-gel immobilized stationary phase with UV absorbance detection. AB - A chromatography column on a chip was fabricated by immobilizing reversed-phase stationary phase particles (5 microm, C4) using sol-gel technology. Channels were fabricated in quartz using photolithography and wet etching. Localization of the stationary phase was achieved by immobilizing the stationary phase at the desired location in the separation channel prior to bonding of the cover plate. Cross channel design was employed for gated injection. An optical fiber setup was developed for carrying out on-chip UV absorbance detection. The effective optical path length was theoretically determined for the trapezoidal shaped channel and the result was shown to match closely with the experimentally determined value. The effect of applied voltage on velocity was evaluated using thiourea as an unretained marker. Separation performance of the stationary phase was demonstrated by separation of three peptides (Trp-Ala, Leu-Trp and Trp-Trp) under isocratic chromatographic conditions. PMID- 15354450 TI - Influence of moderate Joule heating on electroosmotic flow velocity, retention, and efficiency in capillary electrochromatography. AB - The influence of Joule heating on electroosmotic flow velocity, the retention factor of neutral analytes, and separation efficiency in capillary electrochromatography was investigated theoretically and experimentally. A plot of electrical current against the applied electrical field strength was used to evaluate the Joule heating effect. When the mobile phase concentration of Tris buffer exceeded 5.0 mM in the studied capillary electrochromatography systems using particulate and monolithic columns (with an accompanying power level of heat dissipation higher than 0.35 W/m), the Joule heating effect became clearly noticeable. Theoretical models for describing the variation of electroosmotic flow velocity with increasing applied field strength and the change of retention factors for neutral analytes with electrical field strength at higher Tris buffer concentrations were analyzed to explain consequences of Joule heating in capillary electrochromatography. Qualitative agreement between experimental data and implications of the theoretical model analysis was observed. The decrease of separation efficiency in capillary electrochromatography with macroporous octadecylsilica particles at high buffer concentration can be also attributed to Joule heating mainly via the increased axial diffusion of the analyte molecules and dispersion of solute bands by a nonuniform electroosmotic flow profile over the column cross-section. However, within a moderate temperature range, the contribution of the macroscopic velocity profile in the column arising from radial temperature gradients is insignificant. PMID- 15354451 TI - Analysis of ketorolac and its related impurities by capillary electrochromatography. AB - Capillary electrochromatography (CEC) was employed for the assay of ketorolac (KT) and its known related impurities [1-hydroxy analog of ketorolac (HK), 1-keto analog of ketorolac (KK), ketorolac decarboxylated (DK)] in both drug substance and coated tablets. Detection was made at 323 nm and flufenamic acid was selected as internal standard. The experiments were performed in a 100 microm i.d. capillary packed with RP-18 silica particles (33.0, 24.5, 23.0 cm total, effective and packed lengths, respectively). The composition of the mobile phase was optimised by changing pH of the buffer and acetonitrile (ACN) content and by addition of other organic modifiers (methanol, ethanol, isopropanol, n-propanol) in order to evaluate the effect of these factors on the method performance (efficiency, retention and resolution). The optimum mobile phase consisted of a mixture of 50 mM ammonium formate buffer pH 3.5-water-acetonitrile (10:20:70, v/v/v), while voltage and temperature were set at 30 kV and 20 degrees C, respectively. Applying these conditions, all peaks were baseline resolved and the analysis was performed in less than 9 min. Selectivity, repeatability of retention time and peak area, detection and quantitation limits, linearity and range, precision and accuracy were also investigated. R.S.D. and bias values obtained for all the analytes were below 5% and sensitivity was satisfactory, thus the method was deemed suitable for pharmaceutical quality control. Applying the method to coated tablets, a recovery of 98.5+/-0.8% and an R.S.D. of 0.5% were found. PMID- 15354452 TI - Study of flow rate in pressurized gradient capillary electrochromatography using splitter and separation of peptides using an Amide stationary phase. AB - A pressurized gradient capillary electrochromatograph using a splitter was constructed. The variation in flow rate during gradient elution was investigated and separations of peptides using an Amide stationary phase were demonstrated. The flow rate, which is one of the important factors to control chromatographic behavior, was increased during the gradient elution, and the mismatching of mobile phase between the column and the resistance tubing derived three variation patterns in the flow rate. The electrophoretic migration in electrochromatography could enhance in separation of peptides. The separated peak number of tryptic digest of bovine serum albumin was increased from 30 to 40 by the application of +5 kV. PMID- 15354453 TI - Study of the influence of the aspect ratio on efficiency, flow resistance and retention factors of packed capillary columns in pressure- and electrically driven liquid chromatography. AB - The influence of the aspect ratio, rho (rho = column diameter/particle diameter), on column parameters such as efficiency, retention factors and flow resistance was studied in both high-performance liquid chromatography and capillary electrochromatography with packed capillary columns. In order to compare the true efficiencies of different columns, a procedure to account for external band broadening was applied. High efficiencies (reduced plate height h approximately 2) were obtained with capillary columns with internal diameters of 150-, 100-, and 75-microm, packed with 10-microm particles. In contrast to previous reports in the literature, no significant improvements in efficiency or flow resistance were observed when the aspect ratio of such columns was decreased. Our observations suggest that the wall effect in these types of columns is not significant. When the aspect ratio was decreased by increasing the particle size, a decrease in reduced plate height was observed. However, the results of flow resistance measurements showed that the latter effect should be attributed to differences in packing and particle batch quality rather than to differences in the aspect ratio. PMID- 15354454 TI - Separation of polyethylene glycol-modified proteins by open tubular capillary electrochromatography. AB - This study involves the characterization of six polyethylene glycol-modified proteins by open tubular capillary electrochromatography, a high-resolution, versatile and reproducible technique for the analysis of biomolecules and pharmaceuticals. Optimized conditions were obtained with respect to type of capillary modification (cholesterol and octadecyl), applied voltage (+20 and -20 kV), buffer pH (2.14-8.14) and addition of methanol modifier to the mobile phase. Electrochromatograms were obtained with both cathodic and anodic applied electric fields. In the case of one PEG-protein, superoxide dismutase, a comparison was made to a previous study. Reproducibility and column lifetime were also evaluated in assessing the usefulness of the method. PMID- 15354455 TI - Polymer-brush stationary phases for open-tubular capillary electrochromatography. AB - Synthesis of poly(2-hydroxyethyl methacrylate) (PHEMA) brushes from the inside of silica capillaries by surface-initiated atom transfer radical polymerization (ATRP) yields unique stationary phases for open-tubular capillary electrochromatography (OT-CEC). Although PHEMA brushes have only a small effect on the separation of a set of phenols and anilines, derivatization of PHEMA with ethylenediamine (en) allows baseline resolution of several anilines that co-elute from bare silica capillaries. Derivatization of PHEMA with octanoyl chloride (C8 PHEMA films) affords even better resolution in the separation of a series of phenols and anilines. Increasing the thickness of C8-PHEMA coatings by a factor of 2 enhances resolution for several solute pairs, presumably because of an increase in the effective stationary phase to mobile phase volume ratio. Thus, this work demonstrates that thick polymer brushes provide a tunable stationary phase with a much larger phase ratio than is available from monolayer wall coatings. Through appropriate choice of derivatizing reagents, these polymer brushes should allow separation of a wide range of neutral molecules as well as compounds with similar electrophoretic mobilities. PMID- 15354456 TI - Dynamics of some parameters of macula cariosa alba treated with fluoride gel. AB - AIM: The purpose of this study was to follow up the changes of the level of demineralization and the area of white spots during treatment with 0.42% fluoride gel. METHODS: 182 white spots in 27 children (aged 7 to 16) were followed. Two groups were formed - a control group (70 spots) and an experimental group (112 spots). The level of demineralization was assessed by a colour test using 2% methylene blue. Area changes were measured in square millimetres. The initial area was taken to represent 100%. Four applications at 1-week intervals were made. Changes were assessed at the end of months 1, 2, 6, and 12. RESULTS: The colour test showed a decrease in the respective values during the study. The decrease was greater in the experimental group reaching statistically significant difference after the 1st month. There was a slight decrease in the average area of the spots in the control group. At the end of the study the area was reduced to 92.01% of the initial area. The average area in the experimental group at the end of the study was 28.16% of that at baseline. CONCLUSION: These results show the high efficaciousness of the treatment of incipient caries with 0.42% fluoride gel. PMID- 15354457 TI - Esthetic restoration of traumatized permanent teeth in children using composite vestibular veneers (preliminary communication). AB - All people are eager to have attractive dental looks. Esthetic corrections of defects caused by colour alteration are required more and more frequently by dental patients. Very often even the form of viable or devitalised teeth in children is wanted to be changed. The modern composite materials allow this to be done by vestibular veneers satisfying even the highest requirements in this respect. THE AIM of the study was to address the matter of the restoration of traumatised permanent children's teeth by composite veneers. MATERIALS AND METHODS: The study included 30 traumatized permanent anterior teeth in children which, after dealing with the trauma, were restored by means of composite veneers. The teeth were first prepared using a suitable method and the composite veneers were fabricated during the same visit. The patients were evaluated at one, three and six months. RESULTS: The colour, margin integrity, surface structure, and anatomic form were rated using the California Dental Association's criteria. CONCLUSIONS. Restoration of traumatized permanent teeth in children with vestibular veneers yields very good clinical results. The composite veneers is an alternative method in restoration of such teeth in children. PMID- 15354458 TI - Dynamics of caries activity and caries reduction in group prophylaxis with fluoride gel. AB - AIM: The study aims at following up the dynamics of caries activity (As) and caries reduction (Rs) of first permanent molars (M1) in group prophylaxis with 0.42% fluoride gel in groups with relatively low caries risk. METHODS: A three year-long study on 213 7-year olds from Plovdiv was carried out. Children were assigned to two groups - an experimental group and a control group. The children in both groups had no clinical caries on M1 (DMFS = 0) and dft < the means for seven-year olds in Plovdiv. The screening procedure excluded children who were undergoing orthodontic treatment. Eight applications of 0.42% fluoride gel each at 3-month intervals were made in the experimental group. RESULTS: In the first two years caries activity was significantly higher in the control group. In the third year, after discontinuing gel application caries activity in the experimental group slightly increased while that in the control group decreased. No statistically significant differences were found. Caries reduction was highest in the first year (73.12%) with slight decline in the 2nd year (71.12%). In the 3rd year caries reduction was only 17.07%. Despite this, Rs of M1 as measured for the whole period at the end of the 3rd year was 61.93%. CONCLUSION: The first permanent molars are susceptible to caries attack even in groups with relatively low caries risk. The results of this study demonstrate the high effectiveness of prophylaxis using fluoride gel. PMID- 15354459 TI - A study on fear provoking factors in students from the town of Plovdiv. AB - The self-report measure of fear of children dental patients is an important point in screening and determining strategies of behaviour's management. AIM: The aim of this investigation was to assess dental fear in students from the town of Plovdiv, aged 12, 13 and 14, by using Dental Fear Survey Schedule for Children (DFSS-DS) and to determine the causes. MATERIAL AND METHODS: The study included 312 students (161 girls and 151 boys) aged 12, 13 and 14, residents of Plovdiv. They completed the Children Fear Survey Schedule - Dental Subscale. 15 personalities, objects and circumstances have been given estimation points varying from 1 to 5. RESULTS: The results showed that the average level of fear reported amounted to 32.04 +/- 9.8 in girls and to 22.56 +/- 7.6 in boys. Fear of choking is the most frequently reported fear, fear of dentist drilling machines comes next. CONCLUSIONS: Most of the students at the ages of 12, 13 and 14 report quite a fear respect to dental treatment. As a result their oral health gets worse and requires working out of strategies to overcome fear. PMID- 15354460 TI - Measure of efficiency of health education with reference to caries prevention. AB - AIM: The aim of the study was to measure the efficiency of printed health education matters with reference to caries prevention and the endurance of knowledge acquired. MATERIAL AND METHODS: The study included 74 parents of children coming from three kindergartens in the town of Plovdiv. RESULTS: The results showed that the information was received well by the parents (86.2% correct answers). The repeated test after 6 months showed worse results than those from the first test, with an increase of the wrong answers (1st test - 14 wrong and 36 incomplete answers; 2nd test - 29 wrong and 44 incomplete answers). CONCLUSIONS: The information in this form is received well by the parents but requires periodic repetition for the purpose of assimilating the newly-acquired knowledge. PMID- 15354461 TI - A study of dentists' opinion about caries treatment of 1-3- year-old children. AB - AIM: The purpose of the present study was to examine the opinion of dentists from Plovdiv about the problems connected with treating caries of children one to three years of age. METHODS: The study included 129 dentists from Plovdiv. It was carried out by a direct group inquiry. A special questionnaire was prepared. The data obtained were analysed by non-parametrical and graphical analysis. RESULTS: The results showed that 49.60% of the inquired dentists did not treat one- to three-year-old children. 21.70% of them referred to psychological problems among children as a preventive reason; 14.00% - lack of suitable methods; 10.00% felt they were not well-trained in this field and 19.40% answered that they did not take pleasure in working with this group of patients. The greatest difficulties in treating children were met by the dentists general practitioners and those without specialty. CONCLUSION: The results of this study prove the necessity of additional qualification for the general practising dentists together with the development of methods for dental caries treatment that are easy to use in early childhood. PMID- 15354462 TI - A clinical study of a silicate toothpaste with extract from propolis. AB - Propolis is bee-produced substance with pronounced anti-inflammatory effect. It is an ingredient of many drugs; it is added to toothpastes as a prophylactic component for periodontal diseases. The plaque-cleaning, plaque-inhibiting and anti-inflammatory actions of the silicate paste were studied. The study included 42 individuals in good clinical health and a minimum of 20 intact teeth - 11, 16, 24, 31, 36, 46. Control exams were performed on days 1, 7, 14, 21 and 28. The baseline values gradually decreased after each control tooth brushing with a statistically significant difference between them. The toothpaste shows very good plaque-cleaning, plaque-inhibiting and anti-inflammatory effect. PMID- 15354463 TI - Preliminary results from the vector therapy in patients with chronic adult periodontitis. AB - Complete removal of the dental plaque, dental calculus, granulation tissue and necrotic cement from periodontal pockets is of paramount importance for the proper functioning of the regeneration process in the affected periodontium. The purpose of the current study was to obtain preliminary results of the administered vector therapy in patients with chronic periodontitis. Ten patients with 124 multiple-root and 120 single-root teeth were studied; 600 and 480 root surfaces, respectively, were treated. At each appointment we evaluated the oral hygiene of the patient, the presence of plaque-retention zones, bleeding, suppuration, recession, mobility, the probing depth, the quantity and pH of the crevicular liquid, presence of hyperesthesia. After the initial treatment of the root surfaces abrupt improvement of the clinical condition was observed. One month after the therapy there was reduction of the probing depth and the mobility of teeth, less plaque and less bleeding, decreased retention indices, as well as decrease in the crevicular liquid. PMID- 15354464 TI - Root canal filling of endodontically treated teeth by dentistry students in clinical offices from 1996 to 1998--radiological results. AB - THE PURPOSE of our study was to evaluate radiologically the degree of root canal filling of teeth treated by students. METHODS: 724 segment radiographs with an overall number of 1142 canals were examined. The level of root canal filling was measured from the radiographic apex. The objects of the study were divided into three groups: 1. canals filled to more than 2 mm from the apex; 2. canals filled up to 2 mm from the apex; 3. canals filled to the radiographic apex; 4. overfilled canals. The objects from the second and the third group were combined as a group of successfully filled canals. RESULTS: The results showed 62.90% of successfully filled canals, 27.80% of the canals were filled more than 2 mm from the radiological apex and 9.30% - with overfilling. The greatest percentage of canals filled more than 2 mm from the apex refers to the mesial and vestibular canals of the molars, and after them the most frequently obturated canals are those of single-root teeth (57.94%). CONCLUSIONS: The results obtained allow us to estimate the endodontic treatment of the students as good. The greatest difficulties are encountered in filling the vestibular and mesial canals of the molars. PMID- 15354465 TI - Comparative in vitro study of pulp capping materials. AB - AIM: In a comparative in vitro study we examined the bactericidal effect, adhesion and shrinkage of 5% potassium nitrate in Carboxy-Adhaesor, 5% potassium chloride in Carboxy-Adhaesor, Carboxy-Adhaesor, Adhaesor and Dycal. MATERIAL AND METHODS: The bactericidal effect of the investigated pulp capping materials on the isolated microorganisms Staphylococcus aureus, Staph. epidermidis, Streptococcus viridans alpha-haemolyticus, Streptococcus pneumoniae and Streptococcus beta-haemolyticus was assessed by the diffusion method around each disk in mm. The adhesion and shrinkage were assessed after hardening of the material. RESULTS: The results obtained showed higher bactericidal activity of polycarboxylate cements in comparison to Adhaesor and Dycal. The largest zone of inhibition was found around the disks with polyacrilic acid, followed by 5% KNO3 in polyacrilic acid, 5% KCl in polyacrilic acid; significantly smaller is the zone around Dycal. The difference was statistically significant (p<0.01). The adhesion and shrinkage of polycarboxylate cements were estimated as very good, better than those of Adhaesor and Dycal. CONCLUSION: The bactericidal effect of 5% KNO3 in polycarboxylate cement is very good for the microorganisms isolated from deep caries. The adhesion to hard tissues is very good, which suggests good marginal adaptation to the cavity. PMID- 15354466 TI - Electrometric determination of the working length of root canals by means of Apex locator. AB - AIM: To evaluate an apex locator in determining the working length of root canals and detecting micro-communications with the pulp chamber. METHODS: The apparatus was tested on 21 patients, aged 18 - 70, who received endodontic treatment of 34 teeth with 45 root canals. The apparatus was used in compliance with the instructions of the manufacturer. The precision of the instrument was determined by a radiograph of the tooth with a file inserted in it at a depth equal to the length determined with the Apex locator. The obtained results were divided into three groups: canals determined as longer; canals in which the outline of the instrument was close to the roentgenologic apex; and canals determined as shorter. CONCLUSION: The average deviation from the roentgenologic apex was 1.17 mm, which is, in fact, the area of the physiological apex opening - the ideal limits for treatment and obturation. The apparatus can be successfully used to make differential diagnosis is useful of deep caries with pulpitis chronica ulcerosa. It is ergonomic and easy to use. PMID- 15354467 TI - Possibilities for restoration of severely destroyed dental crowns with inlays. AB - In everyday dentistry practice cases with diffuse destruction of masticatory teeth are common. In the study presented the possibilities of their restoration with metal or metal-ceramic inlays are reviewed. Twenty-one inlays were fabricated, 16 from metal and 5 from metal-ceramic. The quality of the fabricated obturations were assessed immediately after they were cemented and one year later. The results showed that this is a reliable method. During the studied period there were no changes in the inlays, the teeth tissues and marginal adaptation. PMID- 15354468 TI - Quantity determination of T- and B-leucocytes in peripheral blood in patients with candida induced subprosthetic stomatitis. AB - INTRODUCTION: The presence of pathological transformation of Candida species and especially of C. albicans is determined to a great degree by the status of humoral and especially of the cell-mediated immunity. The prosthetic stomatitis (Stomatitis subprothetica) and its development also depends on the immunological status of the organism. PURPOSE: In the present study we have set as a purpose to define the quantity of B- and T-lymphocytes (T-early and T-late) in the peripheral blood in patients with subprosthetic stomatitis and isolated and proved C. albicans from the microbial prosthetic plate and the mucosa under the prosthesis. MATERIALS AND METHODS: We defined the quantity of the early and late E-rosette-forming T-lymphocytes by the method of Bach (1973) and The B lymphocytes with complement receptors by the method of Mendes et al (1974) in 69 patients with subprosthetic stomatitis and in 33 people with plate prostheses without stomatitis RESULTS: In both of the studied groups an elevation of the early T-lymphocytes and lower values for the late E-rossette-forming T lymphocytes compared to the normal values were found. The relative proportion of B-lymphocytes is insignificantly higher (p > 0.1) in the patients compared to the control group. The values of T- and B-lymphocytes vary in different directions and they must be interpreted individually. PMID- 15354469 TI - Pregnancy and gingival hyperplasia. AB - The clinical manifestation of plaque-induced gingival inflammation is modulated by the hormonal imbalances during pregnancy. Different forms of severity have been described. During the first months of the pregnancy, the persistent influence of the plaque induces catarrhal inflammation of the gingiva that serves as a base for the development of hyperplastic gingivitis during the last months, modulated by the cumulating hormonal stimuli. In non-controlled cases a development of pyogenic granuloma can be observed. This lesion is rarely observed in women with poor oral hygiene in regions with local irritating factors such as improperly fit obturation or dental calculus and needs specialized treatment. During pregnancy the pyogenic granuloma when treated surgically by excision may reappear due to incomplete excision or non-adequate oral hygiene. The clinical cases presented demonstrate the necessity of prophylaxis and constant control of gingival inflammation during pregnancy and the need of specialized treatment in severe cases. PMID- 15354470 TI - Erythema multiforme--oral manifestations. AB - Modern scientific achievements in the etiology, pathogenesis, diagnosis and treatment of the vesiculobullous lesions of the oral mucosa are of basic significance for the students, who study the diagnosis and the treatment of these diseases, as well as for the clinical practitioners in their everyday practice. The presented new information about the drug-induced or herpes-associated erythema multiforme, the more severe forms - the Stevens-Johnson syndrome and the toxic epidermal necrolysis, is necessary for each practising dentist especially in the diagnosis and treatment of medically compromised patients. Modern investigations confirm the susceptibility of these patients to infections due to primary or secondary immune deficiency. The clinical oral manifestations of erythema multiforme and their treatment are presented. PMID- 15354471 TI - Regenerative therapy of furcation defect. AB - Furcation defects of multiple-root teeth are complications of progressive periodontal diseases. The recommended technique of regenerative therapy in furcation defects of the lower molars is implantation of hydroxylapatite or demineralized freeze-dried bone. A case of a completely absent interradicular bone of 46 tooth in a 35-year-old woman with generalized periodontitis is presented. After endodontic treatment and recovery of the tooth crown we undertook surgical treatment of the bone defect: open curettage, using the technique of Ramfjord and bony reconstruction with aloplastic material bioplant HTR (septodont). The preoperative preparation included clinical prophylaxis and chemical plaque inhibition; postoperatively, rinsing the mouth twice a day with 0.5% metronidazole. The patient was followed up clinically and radiographically up to the 6th month after treatment when the bone structure regenerated. PMID- 15354472 TI - Preparation of abutment teeth with a specialized clinical parallelometer. AB - PURPOSE: A compulsory requirement for the treatment of defects in the dentition is to establish parallelism in the abutment constructions. MATERIALS AND METHODOLOGY: We propose a simplified effective parallelometer that is fixed to the handle of a metal impression tray for both of the jaws. It ensures the same direction of the preparation instruments for all preparation zones. We applied the device for control in clinical practice for preparation of parallel surfaces and profiles. The working models were subjected to comparative analysis with a laboratory parallelometer. DISCUSSION: The results obtained confirm the achieved parallelism between homonymous dental walls. There isn't an index for statistical analysis. CONCLUSIONS: The results achieved allow us to recommend the device for practical application. By using the clinical parallelometer the mistakes in the preparation of bridge abutments will be reduced to a minimum. PMID- 15354473 TI - Comparative evaluation of two commonly accepted tests for adjustment of fixed prosthetic constructions. AB - Regardless of the great advance in prosthetic dentistry related to the development of new more elaborate materials and methods, the dentist must often correct some defects of the newly fabricated prosthetic constructions. In dentistry practice worldwide two tests for adjustment of fixed prosthetic constructions are applied most often - the silicone test and the suspension of Fe2O3 (rouge) and chloroform. The capabilities of the suspension of chloroform and rouge to assess the preliminary contacts between the prosthetic construction and the dental tissues haven't been reviewed in the dentistry literature in Bulgaria. The test is not familiar to the practicing dentists and this is an obstacle for its wide application in everyday practice. The purpose of the current study was to compare the advantages and disadvantages of the silicone test and the modified suspension of halothane and rouge recommended in the recent years. The assessment of the capabilities of the two tests for adjustment is based on the results of their application in the adjustment of crowns and bridges on the working model and on the prosthetic field. The results demonstrate that the two tests have different capabilities in different clinical situations and their concomitant use will allow more precise prosthetic treatment. PMID- 15354474 TI - A device for determination of a plane ensuring parallel preparation. AB - PURPOSE: Parallel preparation of a greater number of teeth with a different tilt is a difficult clinical task. For precise guidance of the preparation instruments with a fixed individual tilt it is necessary to determine the "working preparation plane". A device for orientation and fixation of an individual direction for parallel preparation is created. METHODOLOGY: We have prepared 47 bridge abutments using our methodology. The working models are subjected to control and comparative analysis with a precise laboratory parallelometer. The reported results do not show differences requiring statistical analysis. DISCUSSION: The device serves to determine the working preparation plane. It promotes the fixing of an intraoral parallelometer that controls the achieved parallelism between the homonymous surfaces of dental stumps. PMID- 15354475 TI - Correlation between overcontouring of fixed prosthetic constructions and accumulation of dental plaque. AB - INTRODUCTION: In prosthetic treatment with fixed prosthetic constructions the reproduction of the dimension and form of natural teeth crowns is important for the preservation of normal articulation-occlusal relationship as well as for periodontal health. THE AIM of the study was to evaluate to what degree there is an attempt to preserve and reproduce the natural dental contour in everyday dental practice and its impact on plaque accumulation. MATERIALS AND METHODS: The study was performed in patients with different types of fixed prosthetic constructions - metal, metal-ceramic and the combination of metal/plastic with intact symmetrical homonymous teeth. We used the index of J. Silness and H. Loe (1964) for evaluating the amount of accumulated plaque on the prosthetic construction in the area between the equator of the tooth and gingival margin. RESULTS: Our study reveals that the practicing dentists and dental technicians underestimate the preservation of natural dimension and form of dental crowns. Most often they are significantly enlarged for solving the esthetic requests of the patients. The basis for this is the incorrect preparation of the teeth and the ignorance of the negative effect of overcontouring of fixed prosthetic constructions on the periodontium by the practicing dentists. The overcontouring leads to a greater accumulation of dental plaque in the area between the equator and the margin of the gingiva. PMID- 15354476 TI - The impression as a means for analysis of clinical mistakes in fixed prosthodontics. AB - Earlier and more recent clinical observations show that mistakes made in the early clinical stages of treatment have a negative effect on the time of effectiveness of the prosthetic treatment and its medicobiological value. Finding the most common mistakes and deviations from the clinical and laboratory protocol is of primary importance for creating a qualitative clinical technique and adequate training programs. On the other hand it is difficult for the practicing dentist to perform self-control in his everyday technique for preparation and taking impressions. The purpose of the study was to evaluate the most commonly made clinical mistakes and find how to detect them in time by analysis of the impressions. The study was performed on impressions for crowns and bridges made by generally practising dentists in different regions of the country. The impressions were divided into groups according to manner of their taking - with or without an impression tray. Transverse cuts in planes parallel to the longitudinal axis of the dental stump were used to evaluate the axial inclination and the borders of preparation. The results from the study show that the generally practicing dentists make serious mistakes in the preparation of the teeth for crowns. These mistakes are aggravated by the wrong strategy in taking the impressions. The analysis of the mistakes in the training programs for students and post graduate students is important because it diminishes the chance of their occurring in future. Each dentist can use the impressions taken by him as a means for self-control and evaluation. PMID- 15354477 TI - Study of occlusal-articulation relationships with the help of T-SCAN apparatus. AB - Normal occlusal-articulation relationship between the two jaws ensure the even distribution of the occlusal forces during mastication. Data from the literature and the absence of serious studies evaluating the T-Scan apparatus give reasons for the present study. AIM: To evaluate T-SCAN apparatus for registration and analysis of occlusal contacts in dynamic articulation. MATERIALS AND METHODS: The study was carried out with the medical diagnostic apparatus T-SCAN. Out of 180 students (19-24 years old) examined, we selected 15 with intact dentitions without or with some small obturations and Angle class I relationship between the two jaws. Using T-SCAN we made 7 to 10 records in each of the studied students. RESULTS: The occlusal contacts are presented as two-dimensional and three dimensional images of the contour with different size and colour depending on the force of the pressure. A model of the dental arch is superimposed over the registered occlusal contacts. The place of the centre of the force in central occlusion is determined. The trajectory of the centre of the force during movements of the mandible is recorded which allows determination of the direction and amplitude of movement of the mandible. CONCLUSION: The results from the study reveal that the correct use and analysis of the data from the apparatus T-SCAN provide valuable and reliable information for the localization and distribution of occlusal contacts in dynamic articulation. PMID- 15354478 TI - Histologic study of biocompatibility of the "Cristaloy 7-N" alloy. AB - The most commonly used alloy in dentistry practice in metal-ceramic prosthetic constructions is cobalt-chromium-molybdenum alloy. In Bulgaria the company "Sandental" created a nickel-based alloy for metal-ceramic prostheses. The aim of the study was to asses the medicobiological properties of the nickel-chromium molybdenum alloy "Cristaloy 7-N". The study was performed on eight guinea pigs and four white rats. On both sides of the vertebral column, parallel to it, experimental plates of "Cristaloy 7-N" were implanted on one of the sides and on the contralateral side - golden alloy - .916 gold. After a different period of time - 10, 30, and 90 days, the experimental animals were eutanized and the tissues adjacent to the implanted experimental plates were studied. During the whole period of the study a skin or local or general reaction was not observed. There was no difference in succinate-dehydrogenase activity that is a sign for preserved vital activity. The electron microscopic study of the cells in contact to the implant showed normal ultrastructure of the organelles. The results obtained from the histological, histochemical and electron microscopic study gave evidence for characterizing the alloy "Cristaloy 7-N" as one having high biocompatibility. PMID- 15354479 TI - Measurement of the magnitude of the occlusal forces during articulation. AB - INTRODUCTION: The magnitude and direction of occlusal forces are important factors for the balanced force load of the teeth during articulation. There are no studies available on the magnitude of the occlusal force in the literature. PURPOSE: To analyze the data from the apparatus T-SCAN in order to extend its possibilities in determining specific values of occlusal forces during the process of dynamic articulation. MATERIALS AND METHODS: The apparatus T-SCAN registers occlusal contacts with the help of a sensitive element (sensor) in a net of points. The apparatus scans all these points at specific intervals and depending on the magnitude of the force acting in each one of them enters a number in the range between 0-255 in the memory of the computer. The combination of the points coloured proportionately to the acting force in a given moment form one frame of the film of the occlusion. The sum of all occlusal forces acting in the contact points gives the total occlusal force. RESULTS: The apparatus with the program developed by us showed the following: 1. It shows in numerical and graphic mode the size of the total force of occlusal contacts as well as that one in the left and right half of the dentition or in another zone chosen by the dentist. 2. The size of the area of occlusal contacts is evaluated in a numeric or graphic mode. CONCLUSION: The data for the magnitude of the occlusal forces during the process of dynamic articulation provide valuable information about masticatory load on the teeth or prosthetic constructions. These data can be used in the diagnosis and prosthetic treatment. PMID- 15354480 TI - Teleradiographic method for determining the position of 1:6, 1:4, 1:3, 2:3, 2:4, 2:6 in complete denture. AB - The existing methods for determining the position of 1:3; 2:3 teeth make possible subjective decisions in the construction of complete dentures. The use of objective auxiliary methods and devices that ensure metric determination of the position of 1:6; 1:4; 1:3; 2:3; 2:4; 2:6 haven't been cited in the known literature. PURPOSE: We have set as a purpose to create a methodology that eliminates the subjective determination of the position of 1:3; 2:3 in complete denture construction. METHODOLOGY: With the help of teleradiographic analysis of frontal cephalometry we created a methodology that avoids subjectivity in 1:3; 2:3 preparation. We applied well-known dependencies in orthodontics. - The correlation between the dental arch and the sum of the incisors. - The first law of Izard. The intermolar distance is equal to half the distance measured between the points Z : Z. - The method for graphic depiction of the dental arch of the maxilla created by Hawley and Herbert and refined by Herbst. CONCLUSIONS: The prostheses fabricated according to our methodology with the help of frontal cephalometry provide the possibility to avoid subjectivity in the work of both the dentist and the dental technician. When the frontal teeth are arranged the cutting tooth must lie on the graphic and the side teeth are inscribed in the graphic. The application of frontal cephalometry is an auxiliary method for the arrangement of the teeth in complete dentures, that with its metric values introduces a new aspect in the exactness and the positioning of the teeth in the prostheses. PMID- 15354481 TI - Finite element tension analysis of the supporting tissues of a maxillary canine. AB - INTRODUCTION: The distribution of masticatory load on the teeth and the arising force tensions in them are factors that determine the origine of destructive processes in their periodontium. The development of mathematical models and application of new computer technologies make possible their precise study. They are still not thoroughly studied. PURPOSE: To study the tensions, originating in the periodontium of a canine tooth during occlusal load by the modern method of finite elements (MFE). MATERIALS AND METHODS: A three-dimensional model of a maxillary canine is built by MFE containing 304 finite elements with six varieties of geometrical form, linked in 1409 nodes. It is supposed that the tooth is fixed firmly to the outer surface of the periodontal membrane to the alveolar bone and is loaded in the lingual wall by a force perpendicular to its longitudinal axis and directed from the lingual to the vestibular wall. RESULTS: As a result of the calculations according to MFE the tension state of dental tissues is calculated for diferent degrees of destruction of the alveolar bone. It was established that with the increase of destruction of the alveolar bone for one and the same masticatory load, the tensions in the periodontal membrane also increase. The maximal tensions act in the apex of the root and around the clinical neck of the teeth. CONCLUSION: The results obtained provide precise information of distribution of force tensions in the periodontium of maxillary canines during occlusal load. They serve as a serious theoretical base for future investigations. PMID- 15354482 TI - Broad indications for the application of simultaneous oral-nasal manometering method. AB - The purpose of the study is to enlarge and define the perimeter of application of manometrical method for investigation of nasal breathing. METHOD: We studied patients with congenital or acquired pathological communications of the maxilla as well as patients with perforation of the sinus after teeth extractions that have been treated by prostheses. An original electronic pneumatic device was constructed for our method. RESULTS: Because of the objectiveness of the method and the device we obtained good results in closing the pathological communications in all of the patients. When used in the diagnosis of sinus perforations the method and the device showed the same effectiveness, CONCLUSION: When realization is possible the method guarantees absolute effectiveness in defining leakage of breathed air through pathological communications of the maxilla. With this device, constructed for the study we optimized the application of the method. PMID- 15354483 TI - Application of functional methods for prophylaxis of side effects after prosthetic treatment. AB - The time of appearance and the clinical symptoms of the different periods of functional suitability (FS) and degradation of prosthetic constructions (PC) were studied. It was found that using appropriately chosen methods of functional diagnosis it was possible to foresee the moments when reversible changes in the prosthetic constructions began and when prophylaxis was possible in order to prolong the time of FS of PC, to delay the development of irreversible degenerative changes and to avoid side effects of prosthetics. Knowing the specific clinical manifestations and the time of appearance of different stages of FS it is possible to anticipate the changes that occur on the surface of the prosthesis and the soft tissues at specific moments of the period during which the prosthesis is in the mouth, and determine whether they will progress or not. PMID- 15354484 TI - An appliance for molar distalizatoin with intramandibular stabilization, applied after teeth extraction. AB - The extraoral appliance which is used for the treatment of patients after teeth extraction causes problems when there is lack of cooperation on the part of the patient. The appliance we present here consists of fabricated rings with brackets for the molars, lingual semicannulas, an outer arch, a lingual arch with a palatine button. The appliance distalizes the teeth by the action of NiTi coils placed lingually and vestibularly between the teeth to be distalized. The palatine button, situated behind the frontal teeth, ensures stability of the apparatus. The practical application of the appliance showed good clinical results, which allows us to recommend it in the practice as a method of choice. PMID- 15354486 TI - A method for fabrication of an activator from photopolymer plastic. AB - The authors propose the fabrication of activators from photopolymerizing plastic. The advantages of this method are reviewed in comparison to the classical technology for fabrication of complicated constructions from hydropolymerizing plastic: Lack of residual monomer. Simplification of technology. Significant shortening of the time necessary to obtain the ready apparatus. Easy adjustment and good adaptation. The method is recommended as a method of choice in the practical fabrication of the activator for clinical application. PMID- 15354485 TI - Surgical orthodontic treatment of impacted teeth. AB - The authors demonstrate their clinical experience in the methodology developed by them for surgical-orthodontic treatment of impacted teeth. The material is based on 34 patients - 21 women and 13 men for the period 1994 - 1997. The patients were treated by the following methodology: after surgical exposure of the impacted tooth a photo-composite devise for tooth-alveolar traction is fixed to the crown part of the tooth by photopolymerization. The methodology is illustrated with a case of a patient treated for an impacted canine tooth. PMID- 15354487 TI - Plate osteosynthesis in the treatment of dislocated fractures of the condylar process of the mandible. AB - Condylar fractures present a significant management problem for the surgeon dealing with maxillofacial trauma patients. The main problems in the treatment of the dislocated fractures of the condylar process are: dislocation of the condyle, the delicate operative technique and difficult reposition of the condyle. The authors report a case of fractured and dislocated condylar process of the fabricated mandible which was osteosynthesized by plate. The case is analyzed and the indications for this method are discussed. PMID- 15354488 TI - Reconstruction of the lower lip. AB - INTRODUCTION: There are studies reporting the functional advantages of radiotherapy against surgical treatment. PURPOSE: Because of this we decided to substantiate the surgical treatment as a method of choice in the reconstruction of defects of lower lip after cancer resection. MATERIAL AND METHODS: The author has developed a modification of the method of Bernard for reconstruction of defects of the lower lip after cancer resection. RESULTS: A horizontal incision is made from the angle of the mouth to the cheek, and at the end of the incision a triangle of Burov is cut. After the resection of the tumour, from the lower edge of the defect (the transitional fold), a second incision is done with orientation to the submandibular area at the end of which a second triangle of Burov is cut. The incision takes into consideration the direction of skin folds for achieving better esthetic effect. This refined method was applied in 14 patients with lower lip carcinoma, 7 of them with unilateral flaps and 7- with bilateral flaps from 1994 to 1999. CONCLUSIONS: The functional and esthetic results of the modified method for lower lip reconstruction are better because they lead to better mobility, symmetry of the reconstructed lower lip and optimal wideness of the mouth and the sensitivity is preserved. A mathematical method for planning of the tissues, needed for the replacement of the defect after the excision of the tumour in healthy tissues are under development. PMID- 15354489 TI - Long term results in the treatment of multiple gingival recessions. AB - Exposed root surfaces cause esthetic problems and hypersensitivity to thermal and tactile stimuli. Plastic surgical treatment solves these problems and yields good permanent results. The authors share their experience of the treatment of 30 patients with 186 gingival recessions localized in the lower and upper jaws, frontally and premolarly. The clinico-biometrical studies conducted over a period of 3 to 15 years showed a mean root coverage of 88.48% and in 17 patients the root coverage is 100%. The applied method can successfully deal with multiple gingival recessions in only two visits preserving the periodontium of the adjacent teeth. The surgical technique and the results of its application are illustrated by the results of well-documented cases. PMID- 15354490 TI - Lower lip cancer morbidity in three regions in South Bulgaria for a period of 15 years (1985-1999). AB - INTRODUCTION: In most of the countries lower lip cancer in men varies from 1.0 case per 100 000 (in England) to 8.9 cases per 100 000 (in Hungary). We have set as a PURPOSE: To follow the dynamics in lower lip cancer morbidity in three regions in South Bulgaria. MATERIALS AND METHODS: A cohort study of lower lip cancer is performed for a period of 15 years. RESULTS: In 1985 lower lip cancer morbidity for the three regions was 4.3 cases per 100 000 and in 1999 it was 3.49 cases per 100 000. Morbitdity retains a comparatively high level. In 1985 morbidity in men was 8.5 cases per 100 000 and in women it was 0.7 cases per 100 000. In 1999 the incidence of morbidity in men decreased to 4.9 cases per 100 000, while in women it increased to 2.1 cases per 100 000. The incidence of morbidity rises with age. The factor that most influences morbidity is smoking. A definite role plays also poor condition of the teeth (carious teeth, bad prostheses and crowns, dental calculus). CONCLUSION: The incidence of morbidity in lower lip cancer in men is 4 times as high as that in women. PMID- 15354491 TI - Surgical treatment of squamous cell carcinoma of the lower lip. AB - INTRODUCTION: A controversy still exists regarding the preferable mode of treatment - surgical treatment or radiotherapy of 1st stage squamous cell carcinoma of the lower lip. That is why we have set as a PURPOSE: To compare the two methods of treatment and to propose a therapeutic approach. METHODS: A retrospective longitudinal study of patients with squamous cell carcinoma (SCC) of the lower lip from three regions in South Bulgaria (Plovdiv, Pazardjik and Smolian), who have been treated primarily with surgical operation (n=184) or by radiotherapy (n=592) was performed. RESULTS: Following up 184 patients who have undergone surgical treatment (vermilionectomy) as a primary therapeutic method for a period of 15 years, we have observed favourable results in 92.5% of the cases (n=170). We found local recurrences in 3.1% of the cases (n=6) and regional lymph-node metastases in 4.4% of the cases (n=8). Five-year overall survival rates was 81% and 10-year survival rates - 64%. The corresponding relapse-free five-year survival was 89% and ten-year survival - 82%. The results from the 592 patients treated with radiotherapy are: therapeutically cured - 90.5% of the cases (n=536), local recurrences - 4.3% of the cases (n=25) and regional lymph node metastases (n=31.) CONCLUSIONS: Vermilionectomy is a suitable method for surgical treatment of 1st-stage SCC of the lower lip with a thickness of the tumour up to 3 mm and a size of the primary tumour up to 1.5 cm. PMID- 15354492 TI - Tendencies in the oral cavity cancer morbidity in three regions of South Bulgaria for a period of 15 years (1985-1999). AB - INTRODUCTION: Oral cavity cancer constitutes 4% of all malignancies. In Bulgaria it is 2.21% of all malignancies. PURPOSE: In the present study we have set as an aim to investigate the tendencies of oral cavity cancer morbidity in three regions of South Bulgaria for a period of 15 years. METHODS: A retrospective epidemiological study of oral cavity cancer was performed. RESULTS: The calculated speed of growth is characterized with dynamic growth. Taking 1985 for a base, morbidity increases by 58.6% in 1999. Morbidity in men increased from 8.9 cases per 100 000 in 1985 to 10.12 cases per 100 000 in 1999; in women it increased from 1.4 cases per 100 000 in 1985 to 2.25 cases per 100 000 in 1999. We have found that morbidity in men over the age of 60 has increased more than 25 times and in women - more than 2 times. Mortality varies from 2.64 to 2.78 cases per 100 000. An important risk factor for oral cavity cancer development is poor oral hygiene. CONCLUSIONS: Oral cavity cancer morbidity increases from 3.6 to 8.08 cases per 100 000 in 1999. The risk is 6.9 times higher in men than in women. The risk factors are smoking, alcohol abuse and poor oral hygiene. PMID- 15354493 TI - Clinical and statistical study of salivary gland adenoma. AB - The authors studied 100 patients with salivary gland adenomas for a period of ten years (from 1989 to 1999). Different types of adenomas regarding their glandular localization, clinical presentation, the age and the sex of the patients were studied. Different diagnostic methods were applied: medical history, clinical examination, sonography, scintigraphy, tomodensitometry, sialography, express morphological diagnosis and permanent histological preparation. All patients were treated surgically: in 61% lateral parotidectomy was performed, in 1.4% - conservative parotidectomy and in 24% - enucleation. The authors report about 7.75% recurrence of pleomorphic adenoma and 6% malignization, mainly in the recurrences of the pleomorphic adenoma. In 22 of the patients a postoperative paresis was observed, in 18 of them it was transitory and gradually disappeared after 8 to 9 months. Auriculotemporal syndrome was observed in 4 of the patients. PMID- 15354494 TI - A review of scales for assessment of the quality of life (QOL) in patients with oral cavity cancer. AB - This article is a review of the quality of life (QOL) research in oral cavity cancer patients. The authors set out to carry out this study because research on the problem is still in its infantile period. Most authors use the following scales for assessment: Scale 1 - ability to eat in public; comprehensible speech; normal diet; change in taste, Scale 2 - social adaptation; pain and disfigurement, Scale 3 - determination of the physical and functional status; psychological status; social functioning, Scale 4 - reproduction/pastimes; sleep/rest; home management; work; eating, Scale 5 - fatigue and QOL. One of the indices most frequently used to measure cancer patients' quality of life is the Karnofsky's scale. The Karnofsky's index is determined by two independent factors: the patient oncologist and the nursing team, who perform evaluations on the basis of the patient's response. The differences between these two methods is not statistically significant. Taking into consideration the proposed scales in the literature the authors have prepared a questionnaire which includes a scale for QOL assessment of oral cavity cancer patients to be used in a future survey on the quality of life of cancer patients in some regions of Bulgaria. PMID- 15354495 TI - A denture complication: sarcoid-like foreign body granulomas of the tongue. AB - Sarcoidosis in the oral cavity occurs extremely rare (there have been described only 47 cases in the literature) and is almost always doubtful. We describe a protruding painful nodule (1 cm) on the apex of the tongue of a 65-year-old female, adjacent to the site of irritation of a new denture. Histologically it was confluent mass of non-caseous granulomas, rich in Langhans' cells. Silver impregnation showed them stamped out as in sarcoidosis. Corticosteroids proved to be ineffective and the therapy was discontinued because of the concomitant diabetes. Six months later, similar nodules reappeared on the same place. When examined in polarized light these giant cells were found to be of the foreign body type, each of them containing phagotized particles displaying bright yellow and dark green colour. We assume a sarcoid-like reaction of traumatic origin. There was no recurrence of the condition after removal of the denture. PMID- 15354496 TI - Primary malignant melanoma of oral cavity. AB - Primary oral malignant melanomas (POMM) are very rare. Definitive identification is usually possible only after they develop metastases. So every new case is instructive. P.P., a 76-year-old woman presented with a painless exophytic mass (measuring 3 x 3.5 cm) on a pedicle in the right alveolar crest of the upper jaw (teeth 6, 7 and 8) and the palate. The mass was dark red with lobate and ulcerated surface and dense and elastic consistency. Electroresection was performed until healthy tissue was reached. One year later there were enlarged lymph nodes submandibularly on the right: a metastasis arising from an achromatic melanoma of the upper jaw. Fontana-Masson staining, expression of S-100 protein in the melanocytes. The patient was in good condition two years after the administered telegamma and immunotherapy. POMM affects mainly men at advanced age. It is four times as frequent in the upper jaw and palate as it is on the surfaces. The tongue is not affected by POMM but is targeted by the melanoma metastases. Only one third of all cases of POMM are pigmented. They arise on the basis of premelanotic changes. Every equivocal pigmented lesion in the oral cavity should be obligatorily subjected to biopsy. Primary lymph dissection is not used routinely. The therapy of choice is a radical resection combined with chemical, radio and immunotherapy. Prognosis remains grave in spite of treatment. PMID- 15354497 TI - Doctor-patient relationships in global society. Informed consent in dentistry. AB - The concept for initiation of treatment only after the explicit consent of the patient, based on preliminary information, is the most important element determining the relationship between patient and doctor nowadays. The application of this concept in dentistry and its inclusion in the professional documents regulating these relationships needs more comprehensive and modern conditions relevant analysis of ethical, legal and professional aspects of the problem. The purpose of the study was to define the modern view of informed consent and its application in dentistry in different social environment. The general and specific features of the evolution of the problem and the social practice in Bulgaria, The European Community and Northern America are discussed in the context of the global tendency for free movement of patients and mutual recognition of professional qualification. The results suggest that despite the different degree of social and economical development in different countries the interest in the problem grows significantly and harmonization of legislation for health protection is based on the new social decree in the construction of doctor patient relationships. It is expected that the comparative studies in this field will promote the improvement of dentists' training and improvement of scientific and expert exchange in solving problems of patents' rights and professional ethics. PMID- 15354499 TI - More thoughts on the doctorate degree for NPs. PMID- 15354498 TI - A gut feeling. PMID- 15354500 TI - India has high-tech health care, too. PMID- 15354501 TI - The basics of business plans. PMID- 15354502 TI - Say what you mean. PMID- 15354503 TI - Polycystic ovary syndrome, obesity and insulin resistance: the new female triad. PMID- 15354504 TI - Dietatry guidelines for type 2 diabetes. A toolbox for nurse practitioners. PMID- 15354505 TI - Technologic advances in glucose monitoring. New systems offer more in-depth information. PMID- 15354506 TI - Diabetes self-management education. Achieving success in clinical settings. PMID- 15354507 TI - Perplexing pediatric emergencies. Fourteen scenarios to fine-tune your diagnostic skills. PMID- 15354508 TI - Rape and sexual assault. Understanding the offense and the offender. PMID- 15354509 TI - Patient information. Acne: clearing the most common skin condition. PMID- 15354511 TI - Has it really been 28 years? PMID- 15354510 TI - Bioidentical hormone therapy for menopausal and perimenopausal patients. PMID- 15354512 TI - [Progress in the studies of alpha1-receptor blocker for benign prostatic hyperplasia]. AB - Benign prostatic hyperplasia (BPH) is a common senile disease, and its main clinical manifestation is lower urinary tract symptom (LUTS), which has long been afflicting old male patients. Previous study showed that alpha1-receptor in the prostate was involved in the development of LUTS. At present, alpha1-receptor blocker is generally accepted as a choice drug for treating BPH and relieving LUTS. The article reviews the tissue distribution of alpha1-receptor and clinical application of alpha1-receptor blocker. PMID- 15354513 TI - [The effects of interleukin-10 and -8 in chronic prostatitis]. AB - OBJECTIVE: To investigate the effects of interleukin-10 (IL-10) and IL-8 in the pathogenesis and diagnosis of chronic prostatitis (CP). METHODS: Twenty-nine different types of CP patients with complaints of low urinary duct symptom were selected at random, and the diagnosis of CP was established by inquiry of the history of prostatic disease, digital rectal examination of the prostate, and analysis of express prostatic secretions (EPS) for leukocytes. Bacterial infection of the prostate was detected by pre- and post-message test (PPMT) in some of the inflammatory CP patients. Quantitative evaluation of IL-10 and IL-8 levels in the limited voided bladder 3 (VB3) was performed by ELISA methods. Eleven healthy men without CP were chosen as control. RESULTS: The level of IL-10 [(47.1 +/- 4.5) pg/ml] in VB3 in 8 patients with apparent clinical symptoms was significantly higher than in the 11 infertile males [(40.8 +/- 5.7) pg/ml] and the 21 patients with both infertility and asymptomatory inflammatory prostatitis (CP type IV) [(42.7 +/- 6.7) pg/ml], P < 0.05. The levels of IL-8 in the 8 patients with apparent clinical symptoms [(1386.2 +/- 852.6) pg/ml] and in the 13 patients with both infertility and asymptomatory inflammatory prostatitis (CP type IV) [(1203.8 +/- 807.8) pg/ml] were significantly higher than in the 7 healthy males [(412.1 +/- 217.2) pg/ml], P < 0.05. CONCLUSION: IL-10 and IL-8 had a very important role in etiology of CP, and could be detected in VB3 as well as in routine semen and EPS. PMID- 15354514 TI - [Transurethral bipolar plasmakinetic prostatectomy for benign prostatic hyperplasia]. AB - OBJECTIVE: To evaluate the effect and complications of transurethral plasmakinetic prostatectomy (TUPKP) in the treatment of benign prostatic hyperplasia (BPH). METHODS: All 313 patients underwent TUPKP, and the operative indexes and perioperative blood indexes were recorded. After operation, 290, 288 and 142 cases of BPH were followed up at 1 month, 3 months and 1 year respectively. Qmax, IPSS and QOL were measured in all the catamneses. RESULTS: The operative time was (51 +/- 22) min; the mount of blood loss was (66 +/- 60) ml; no TURS occurred in any cases. The mean catheterization time was (11 +/- 10) h and the mean postoperative stay was (3.6 +/- 1.3) d. Qmax increased from (9.0 +/- 4.4) ml/s to (20.5 +/- 7.1) ml/s at 1 month, (21.8 +/- 5.4) ml/s at 3 months and (21.4 +/- 6.6) ml/s at 1 year after operation (P < 0.01). Correspondingly, IPSS decreased from (26.2 +/- 5.1) score to (6.0 +/- 9.0) score, (5.6 +/- 0.8) score and (4.4 +/- 2.7) score (P < 0.01), and the QOL of all the catamneses significantly improved. CONCLUSION: TUPKP, a safe and effective method with fewer complications, can be recommended for the treatment of BPH. PMID- 15354515 TI - [Detection and potential function of anti-FSH antibodies in patients with spermatogenesis dysfunction]. AB - OBJECTIVE: To detect the anti-FSH antibody using ELISA, and further probe into the role of anti-FSH in infertile patients. METHODS: The anti-FSH antibody was detected using ELISA in the serum of patients with spermatogenesis dysfunction, of infertile patients with normal sperm density and motility, and of normal fertile males. RESULTS: The positive rate of anti-FSH antibody in the patients with oligospermia and/or asthenospermia [22.4% (22/98)] was significantly higher than that in the normal fertile [4% (2/50)] (P < 0.05) and that in the infertile patients with normal sperm density and motility [6.7% (2/30)] (P < 0.05). The positive rate of anti-FSH antibody in the patients with oligospermia and/or asthenospermia was lower than that in the patients with azoospermia [54.5% (12/22)] (P < 0.05). There was no significant difference in the positive rate between the normal control and the sterile males with normal sperm density and motility. CONCLUSION: The anti-FSH antibody may be an important factor to cause spermatogenesis dysfunction by combining FSH to form immune compound and depress the activation of FSH. PMID- 15354516 TI - [Four regions 12-core systemic prostate biopsy for diagnosis of prostate cancer]. AB - OBJECTIVE: To evaluate the clinical value of the four regions 12-core systemic prostate biopsy. METHODS: Ninety-one patients suspected of prostate cancer received four regions 12-core systemic prostate biopsy, and the detection rate and complications were compared with those of the sextant method. RESULTS: More positive results were obtained in detecting prostate cancer with the four regions 12-core method, and the complications were similar to those of the sextant biopsy. CONCLUSION: The four regions 12-core biopsy was suggested for patients suspected of prostate cancer with enlarged prostate gland above 40 ml. PMID- 15354517 TI - [Investigation of infection risk and the value of antibiotic prophylaxis during transrectal biopsy of the prostate by endotoxin determination]. AB - OBJECTIVE: To clarify the value of endotoxin determination in evaluating the infection risk and antibiotic prophylaxis during transrectal biopsy of the prostate. METHODS: Forty-eight patients were divided into four groups randomly. Group A (12 patients) were not treated before biopsy. Group B (13 patients) received antibiotic prophylaxis therapy. Group C (12 patients) underwent clusis. Group D (11 patients) received combined treatment. Urine and blood samples of the patients were obtained before and after prostatic biopsy. All samples were cultured for bacteria and investigated for endotoxin concentration by limulus quantitative azo color test. RESULTS: No significant difference in either serum endotoxin or blood bacterial cultures was noted before and after prostatic biopsy (P < 0.05) in all the groups. A significant increase was observed in urine endotoxin after biopsy compared with that before biopsy in Groups A and B (P < 0.05). There was no significant increase in urine endotoxin after biopsy compared with that before biopsy in Group C and D. The positive incidence of urine bacterial culture was significantly increased (P < 0.05) in patients of Group A and B. CONCLUSION: Circulation infection risk following prostatic biopsy was low. Changes of urinary infection were fewer in patients who had undergone clusis and/or antibiotic prophylaxis than in those who had received no or only antibiotic prophylaxis therapy. Endotoxin determination in urine is a reliable, sensitive and simple method for diagnosis of bacterial infection in patients undergoing transrectal biopsy of the prostate. PMID- 15354518 TI - [Questionnaire and pad-test in assessment of incontinence after radical retro pubic prostatectomy]. AB - OBJECTIVE: To confirm urinary incontinence after radical retro-pubic prostatectomy by questionnaire and pad-test, observe the influence of radical prostatectomy on the patient's quality of life (QOL), and study the effect of urethral sphincter preservation and the patient's age on incontinence. METHODS: Questionnaire and pad-test were conducted in 165 consecutive prostate cancer patients who underwent radical prostatectomies. The mean follow-up time was 13 months (12-14 months). Each patient was kept in contact with our incontinence advisor by telephone or direct interview. At 12 months after operation, the patients were asked to the clinic to fill in the questionnaire and underwent the pad-test. In accordance to different operative techniques, the patients were divided into a sphincter repairing group (19 cases) and a sphincter preservation group (146 cases). RESULTS: All the 165 patients were kept in contact with us, underwent the pad-test and filled in the questionnaire. Of the 7 patients diagnosed as incontinence for admitting using pads, only 1 had urinary leakage, and the other 6, who did not use pads very often, showed slight change of QOL. In the pad-test, 5 patients were considered to be incontinent since the pad weight gained > 1 grams. Between the questionnaire and the pad-test there was a high concordant rate (98.8%). Within 3 months after operation, younger patients seemed to return to continence sooner. The continence rate at 12 months after operation was 99.3% in the sphincter preservation group and 94.7% in the sphincter repairing group. The incontinence rates at the removal of the urinary catheter after operation were 60% and 82% in patients aged 51-60 and 61-70, respectively. At 3 months after operation, the incontinence rates were similar in both groups. CONCLUSION: Both the questionnaire and the pad-test are recommendable in the assessment of post-operative incontinence since it can accurately document patients' incontinent status. Preservation of the urethral sphincter and its possible innervations can improve the incontinence rate. Younger patients seem to return to continence sooner than the elders. PMID- 15354519 TI - [Sexual development in adolescents after surgical treatment for cryptorchidism]. AB - OBJECTIVE: To investigate the sexual development in adolescents after surgical treatment for undescended testes. METHODS: One hundred and eighty-four adolescents undergoing surgery for cryptorchidism before the age of 10 years, 22 cases received no surgical management for unilateral undescended testes, and 25 normal controls were studied. The pubic stage, the natural length and girth of the penis, the volume of the testis, serum luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone (T) were examined to find out the effect of age, the location of the testis and operative procedures on prognosis. RESULTS: The pubic stage, the length and girth of the penis, the volume of the testis and T decreased, but FSH and LH increased significantly in 18 adolescents who had undergone bilateral orchiopexy. The volume of the unilaterally undescended testis was significantly smaller than that of the normally descended contralateral gonad in 152 adolescents who had undergone unilateral orchiopexy. FSH was significantly higher in the adolescents surgically treated for unilateral cryptorchidism. Those treated by unilateral orchiectomy presented significantly higher levels of FSH than those by unilateral orchiopexy. The pubic stage, the length and girth of the penis, the volume of the testis and T were significantly higher, but FSH lower in the adolescents treated before the age of 5 years than at the age of 5 or older. The decrease in testicular volume was significantly greater in adolescents with intra-abdominal testes. Significant negative correlation was found between FSH and testicular volume. CONCLUSION: Leydig cell function seems relatively resistant to the hostile environment of the cryptorchidism. Early diagnosis and management of the undescended testis are needed to preserve fertility. PMID- 15354520 TI - [Molecular epidemiologic investigation of infertile male's semen infected by Gardnerella vaginalis]. AB - OBJECTIVE: To investigate the status of the semen of the infertility patients infected by Gardnerella vaginalis (Gv). METHODS: Semen samples from 373 clinic patients of infertility and vaginal samples from 63 positive patients' wives were collected from April 2002 to May 2003. And the samples were tested by nested polymerase chain reaction (nPCR). RESULTS: The positive rate of the infertile males' semen infected by Gv was 44.2%, while that of the postive patients' wives was 87.3%. CONCLUSION: The positive rate of the infertile male's semen infected by Gv is high and Gv can be spread by sexual intercourse. PMID- 15354521 TI - [Exploratory study on related factors of sexual dysfunction among breast cancer patients]. AB - OBJECTIVE: To investigate the factors related to sexual dysfunction among breast cancer patients so as to improve the prevention and treatment of the disorder as well as the life quality of the patients. METHODS: Sixty-five breast cancer patients during the rehabilitation period were interviewed by questionnaire on the sexual function before and after treatment. RESULTS: Age and perception of sex were two important factors for the significant difference in the rate of sexual dysfunction among the patients. In the groups of 45-55 and 56-65 years, the rates of sexual dysfunction were 66.7% and 73.9%, respectively. Compared with the < 45-year group (33.3%), the findings were statistically significant (P < 0.01), and the difference was statistically significant between the incorrect perception group (70.3%) and the correct one (47.6%) (P < 0.05). Of all the factors analyzed in the research, the stage of cancer, treatment methods, vaginal dryness, decreased libido, dyspareunia and sex perception had significant correlation with newly developed sexual dysfunction (P < 0.05). CONCLUSION: The stage of cancer, treatment methods, sex perception, vaginal dryness et al had significant correlation with sexual dysfunction of breast cancer patients after treatment. To treat and prevent sexual dysfunction among breast cancer patients, oncology professionals should initiate communication about sexual difficulties, perform comprehensive assessments, and educate and counsel patients about the management of these difficulties. PMID- 15354522 TI - [Semen analysis of DNA fluorescence detection and two other routine analyzing methods]. AB - OBJECTIVE: To compare semen analysis system of computer-aided sperm analysis (CASA) dyeing with DNA fluorescence (simplified as fluorescence CASA) with two other routine semen analyzing methods. METHODS: Twenty-two male infertile patients were selected at random as research subjects, and semen analysis was performed with fluorescence CASA, grey CASA, and microscopic analysis. RESULTS: Fluorescence CASA could distinguish sperm from non-sperm granules, as well as living from dead sperm. Compared with the sperm density detected by fluorescence CASA, the disparities of sperm densities detected by microscopic analysis and grey CASA were (9.23 +/- 8.01) x 10(6)/ml and (10.27 +/- 6.22) x 10(6)/ml, and the disparity rates of sperm density were 49.06% +/- 49.87% and 43.39% +/- 25.56% respectively. CONCLUSION: Fluorescence CASA was more conformable to the requirements of latest WHO laboratory manual for the examination of human semen, and worth recommending for the clinical analysis of semen features, especially sperm density. PMID- 15354523 TI - [Immunohistochemical localization of attractin in the testis and epididymis of mature male rats]. AB - OBJECTIVE: To localize the attractin protein in the testis and epididymis of mature male rats. METHODS: Testes and epididymides were obtained from mature male Sprague dawley rats (n = 20). Tissues were fixed and prepared for immunohistochemical (IHC) and indirect immunofluorescent (IIF) assay, carried out with antiserum against rat attractin. RESULTS: In the testis of the male rat, there was distinct immunopositive staining on cell membrane and cytoplasm within Leydig cells, primitive spermatogonia, primary spermatocytes, spermatids, Sertoli cells, and peritubular myoid cells. In the epididymis, including caput, corpus, cauda, there was no definitive immunopositive staining within the efferent ductule and epididymal duct. CONCLUSION: Attractin is expressed in the male rat reproductive system and localized within Leydig cells and germ cells. It may be invoved in acting on the reproductive system. And its physiological function has yet to be further studied. PMID- 15354524 TI - [Effect and mechanism of alpha1-adrenoceptor blocker combined with antibiotics for chronic prostatitis]. AB - OBJECTIVE: To investigate the effect and mechanism of alpha1-adrenoceptor blocker combined with antibiotics in the treatment of chronic prostatitis. METHODS: Eighty patients with chronic prostatitis were divided into two groups, one treated with alpha1-adrenoceptor blocker (Terazosin 2 mg qn) and Levo-ofloxacin (0.2 bid), and the other given Levo-ofloxacin (0.2 bid) alone for 6 weeks. Chronic prostatitis symptom index (CPSI), urodynamic data and prostatic secretion examination were compared before and after treatment. RESULTS: The CPSI score of the treated group decreased from 31.8 +/- 7.4 to 15.5 +/-6.6, while that of the control group decreased from 30.9 +/- 7.1 to 21.4 +/- 6.2. There was significant difference between the two groups (P < 0.05). The maximum flow rates before and after the combined treatment were 16.5 +/- 6.3 ml/s and 20.4 +/- 4.6 ml/s, while those before and after Levo-ofloxacin administration were 16.1 +/-5.8 ml/s and 17.3 +/- 6.8 ml/s. The difference was significant (P < 0.05). The maximum urethral pressure of the combined treatment group decreased from 92.5 +/- 15.3 cm H2O to 72.5 +/- 13.4 cm H2O, while that of the control group decreased from 93.2 +/- 14.8 cm H2O to 91.7 +/- 13.6 cm H2O. CONCLUSION: Alpha1-adrenoceptor blocker can lower the intraurethral pressure, which prevents urine from refluxing to the prostate. Alpha1-adrenoceptor blocker combined with antibiotics is effective for chronic prostatitis. PMID- 15354525 TI - [Study of the effect of super painkiller on the sexual function of male rats with type I diabetes]. AB - OBJECTIVE: To study the effects of super painkiller on the sexual function of male Wistar rats with diabetes. METHODS: Thirty-two Wistar rats at 3-5 months of age were divided into two groups at random: 22 in the diabetes group, and 10 in the control group. After 72 hours, the former was further divided into 2 subgroups: non-treatment and super painkiller treatment. In 5 weeks, blood glucose was determined. After the apomorphines experiment, the rats were killed, blood taken from the vein, homogenate prepared from the isolated testis tissue and the level of NO and NOS in the serum and tissue homogenate surveyed, separately. RESULTS: Compared with the control group, serum NO and NOS of the diabetic rats dropped sharply. Compared with the non-treatment group, the serum NO and NOS of the super painkiller treatment group were very high and the difference was significant (P < 0.01). Apomorphine injection showed that the times of penis erection of the treated rats were more than those of the non treatment group (P < 0.01) , but the difference was not significant compared with the control (P > 0.05). CONCLUSION: Super painkiller has sure effect of reducing blood glucose, with a certain curative value for sexual and reproductive malfunction of diabetic rats. PMID- 15354526 TI - [One case report of clitoral priapism and literature review]. AB - OBJECTIVE: To investigate the clinical characteristics of female clitoral priapism. METHODS: A 29-year-old case of painful priapism of the clitoris lasting 60 hours was reported. The etiology, diagnosis and treatment of clitoral priapism were discussed and the literature reviewed. RESULTS: The patient was cured by intracavernous administration of adrenaline. CONCLUSION: Clitoral priapism is extraordinarily rare. It is primarily due to oral antidepressant drugs or pelvic malignant neoplasm which leads to tissue infiltration of the clitoral veins and obstruction of the clitoral corporeal blood flow. The efficient therapeutic approach to clitoral priapism is intracavernous administration of alpha-agonists similar to the treatment of penile priapism. PMID- 15354527 TI - [Investigation of correlative factors affecting successful intrauterine insemination]. AB - OBJECTIVE: To investigate the main correlative factors affecting the pregnancy rate of intrauterine insemination (IUI) by means of analyzing 186 patients and 216 cycles of IUI. METHODS: Comparisons were made between different pregnancy rates from five respects: infertility reasons, stimulation and natural cycles, different protocols for stimulating ovulation (CC group, CC + hMG group, hMG group), two methods for semen treatment (swim-up and gradient centrifugal), and insemination time (endometrial thickness and E2 levels at the day of hCG injection, and times of IUI). RESULTS: The pregnancy rates resulting from male and cervix factors (29.82%, 25.00%) were significantly higher than those from other factors. There were few statistical differences in the pregnancy rates between stimulation and natural cycles (19.54% vs 16.67%), between the three different protocols for stimulating ovulation (CC 6.50%, CC + hMG 14.28%, hMG 22.30%), between gradient centrifugal and swim-up methods (20.13% vs 16.13%) and between one-time IUI and two-time IUI (19.73% vs 17.39%) (P > 0.05). However, the pregnancy rates were statistically correlated with the endometrial thickness at the day of hCG injection and different E2 levels of the follicles over 14 mm in diameter (P < 0.05). CONCLUSION: IUI is of more applied value for infertility caused by male and cervix factors. With these factors in view, to choose the most suitable insemination time is the key to increasing pregnancy rate. PMID- 15354528 TI - [Study of zinc in protecting sperm from sodium nitroprusside damage]. AB - OBJECTIVE: To study the function of zinc in preventing human sperm from being damaged by sodium nitroprusside (SNP), an external NO donor. METHODS: Analyses were made of the function of zinc in protecting sperm from being influenced by SNP in such aspects as sperm motility, head-tail connection and the breakage of sperm DNA chain by using phase-contrast microscope and single cell gel electrophoresis (SCGE). RESULTS: Sperm motility was obviously inhibited by SNP. The percentage of comet cells increased significantly but the stability of sperm head-tail connection decreased. Zinc could promote sperm motility, protect the DNA chain and prevent the sperm head-tail connection from breaking. CONCLUSION: Zinc can protect sperm from being damaged by NO. Its mechanism may be related to the mercaptol group of sperm chromatin. PMID- 15354529 TI - [Advances in studies of male reproductive toxicity of pesticides]. AB - The present article reviews the advances in the studies of male reproductive toxicity of the pesticides that are widely used and commonly researched in the recent years by means of animal experiment and human investigation. The mechanism of male reproductive toxicity of pesticides is discussed. And problems are raised in the evaluation of human reproductive hazards. PMID- 15354530 TI - [Phospholipase C and reproduction]. AB - Phospholipase C (PLC) is a key enzyme in phosphatidyl inositol turnover during signal transduction. The 12 mammalian PLC isozymes identified to date can be divided into five subtypes, beta-type, gamma-type, delta-type, epsilon-type and zeta-type, with extensive difference in structure, regulatory mechanism and tissue distribution. PLC plays important roles not only in sperm acrosome reaction but also in egg activation. The present studies are reviewed on the structure, regulation and function of PLC, especially its function in male reproduction, including triggering Ca2+ oscillations in eggs to activate the eggs and helping embryo development. And the prospect of the clinical application of PLC is discussed. PMID- 15354531 TI - [Prospects for early diagnosis of prostate cancer by detection of plasma DNA]. AB - A major problem for the early diagnosis of prostate cancer is the lack of clinically useful tests for screening a preclinical and asymptomic population without resort to invasive diagnostic procedures. Recent studies have demonstrated the possibility to detect genetic alterations in plasma or serum DNA from patients with prostate cancer or other cancers. Quantification and molecular event are associated with advanced stages and circulating tumor cells. These results indicate a new approach to the early diagnosis and monitoring of prostate cancer by non-invasive screening procedures based on the analysis of genetic changes in plasma. PMID- 15354532 TI - [Female prostatitis]. AB - Female urethral syndrome is very common, but its etiology is not yet known completely and its management is somehow difficult. Up to now, there have been quite a number of reports about the female prostate, whose inflammation is possibly the major cause of female urethral syndrome. PMID- 15354533 TI - Evaluation of certain food additives and contaminants. AB - This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, with a view to recommending acceptable daily intakes (ADIs) and to prepare specifications for the identity and purity of food additives. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of food additives (including flavouring agents) and contaminants, assessments of intake, and the establishment and revision of specifications for food additives. A summary follows of the Committee's evaluations of toxicological and intake data on various specific food additives (alpha-amylase from Bacillus lichenformis containing a genetically engineered alpha-amylase gene from B. licheniformis, annatto extracts, curcumin, diacetyl and fatty acid esters of glycerol, D tagatose, laccase from Myceliophthora thermophila expressed in Aspergillus oryzae, mixed xylanase, beta-glucanase enzyme preparation produced by a strain of Humicola insolens, neotame, polyvinyl alcohol, quillaia extracts and xylanase from Thermomyces lanuginosus expressed in Fusarium venenatum), flavouring agents, a nutritional source of iron (ferrous glycinate, processed with citric acid), a disinfectant for drinking-water (sodium dichloroisocyanurate) and contaminants (cadmium and methylmercury). Annexed to the report are tables summarizing the Committee's recommendations for ADIs of the food additives, recommendations on the flavouring agents considered, and tolerable intakes of the contaminants considered, changes in the status of specifications and further information requested or desired. PMID- 15354534 TI - [DNA protective activity of natural and synthetic antioxidants]. AB - Free radicals attack cell genome in oxidative stress conditions accompanying many human diseases. Mutagenic and carcinogenic xenobiotics cause oxidative DNA damages also. Oxidative DNA damages become intensive in aging organisms. The data on natural and synthetic antioxidants protecting DNA from oxidation are presented in this review. PMID- 15354535 TI - [Electrochemical reduction of cytochrome P450 as a way for construction of biosensors and bioreactors]. AB - The present review describes the data on electrochemical reduction of cytochrome P450. Three generations of enzyme biosensors have been considered. The concept and potentialities of enzyme electrodes--transducers--as the main element on construction of electrochemical biosensors are described. Different types of electrodes for bioelectrochemistry are presented. New experimental approaches for immobilisation of cytochrome P450 based on nanotechnology are reported. Nanobiotechnology in electrochemistry has potential application for production of biosensors and bioreactors for medicine PMID- 15354536 TI - [Induction of cytochrome P450 2A5 in transplanted mouse hepatoma]. AB - We have studied selective induction in vivo isoform of cytochrome P4502A in mouse hepatomas. Activity of coumarin 7-hydroxylase was increased in hepatoma 61 following pyrazole and cobalt chloride treatment. Microsomes isolated from hepatoma 61 transplanted to mice treated with either pyrazole or cobalt chloride catalyzed oxidation of coumarin and 7-ethoxycoumarin at rates 2-2.5-fold higher than in saline controls. Western blot analysis of hepatoma microsomes showed that the increase in functional activity of coumarin 7-hydroxylase was due to induction of CYP2A5 (cytochrome P450 isoenzyme catalysing coumarin 7 hydroxylation). Pyrazole or cobalt chloride induced the enzyme activity in hepatoma 61, whereas we did not measure induction of CYP2A5 in hepatoma 60. The changes in the amount of CYP2A5 in liver were more pronounced after pyrazole treatment than that after cobalt. It is suggested that hepatomas 60 and 61 are originated through initiation of hepatocytes which are localized within different regions of liver lobule. PMID- 15354537 TI - [Use of restriction endonucleases Bst2U I and Acc65 I to detect Kpn I polymorphism of NAT2 gene]. AB - RFLP-analysis was made for 30 human DNA samples. The ability of application of restriction endonucleases Acc65I and Bst2UI to find point mutation C481T in NAT2 gene has been demonstrated for the first time. Variants of these enzymes application are discussed. PMID- 15354538 TI - [Dose-dependent effects of morphine hydrochloride on glycolysis in the rat liver]. AB - The effect of acute morphine intoxication (10, 20 and 40 mg/kg body weight) on the functioning of glycolysis in the rat liver was studied. The most pronounced effect was noted at the dose of 10 mg/kg. It included activation of key glycolytic enzymes: hexokinase, glucokinase and phosphofructokinase and increase blood and liver tissue glucose being increased. The elevated morphine dose did not change glycolytic enzyme activities, decreasing glucose and glucose-6 phosphate contents. PMID- 15354539 TI - [Chemiluminescence intensity and activity of the antioxidant system in dynamics of experimental carcinogenesis combined with chlamydial infection]. AB - Experiments on rats with inoculated sarcoma-45 combined with chlamydial infection showed activation of processes of lipid peroxidation already at early free radical stage of lipid peroxidation, that resulted in subsequent increase of activity of key antioxidant enzymes: superoxide dismutase and catalase. With progression of the tumour process developing under conditions of chlamydial infection the intensity of the increase of enzyme activity decreased. PMID- 15354540 TI - [Cooperative characteristics of androgen receptors]. AB - Molecular characteristics of the androgen receptor (AR) from 6-months old albino rat testes were studied. Using radioligand methods, gel filtration on Toyopearl HW-55F column and analyses of specific testicular cytosol 5-alpha dihydrotestosterone binding in Scatchard and Hill plots it was shown that functional androgen receptor complexes (ARC's) are dimers exhibiting positive cooperativity during hormone ligand binding. The cooperative properties of AR were stable not only at 4 degrees C (Hill coefficient = 2.11) but also at physiological temperature of 36 degrees C (Hill coefficient = 1.95). Presence of Na2MoO4 x 2H2O (10 mM) and/or dilution of cytosols reduced Hill coefficient to 1.03. Addition of morning (but not afternoon) blood sera microquantities (1:20) to monomerized AR preparations resulted in the AR dimerization and restoration of their positive cooperative properties for hormone binding. The effect of morning sera persisted through their pretreatment (15-h at 4 degrees C) with suspensions of 2.5% dextran coated charcoal. Sera from 28-months old rats lacked AR dimerizing activity. PMID- 15354541 TI - [New derivatives of triazino- and imidazoindole with hepatoprotective activity]. AB - The effect of 30 indole derivatives (mainly condensed) containing a thio carbamide fragment was investigated in rats using a model of carbontetrachloride induced hepatitis and a model of partial hepatectomy (for some compounds). Among indoles, 1,2,4-triazino[5,6-b]- and [6,5-b]indole as well as imidazo[4,5-b]indole series the compounds, decreasing concentration of alanine aminotransferase, aspartate aminotransferase and total bilirubin in blood serum and reducing hexenal sleep duration were found. The compounds exceeding in this respect riboxine, potassium orotate and essentiale (used for comparison) and posessing sufficient therapeutic index were found. The correlation between biological activity and ionisation constants was observed for triazinoindole derivatives, while for some indices of liver state the correlation with ionisation constants and distribution coefficients in octanol/water system was found as well. High activity was most probable at pKa < or = 8. For aminoallcyl thioderivatives of triazinoindole the protective effect increased on substituting the aminogroup for hydrogen atom at 8-position, on increasing the side chain length from 2 to 3 methylene links and on replacing of acyclic aliphatic amines residues in a side chain by cyclic amines--pyrrolidine, N-methylpiperazine and morpholine rather than piperidine which increased toxicity. There was no correlation between hepatoprotective and antihypoxic activity while for actoprotectors the probability of liver protection was increased. The results indicate perspectives in searching new hepatoprotective agents among the indicated series of indole derivatives. PMID- 15354542 TI - [The influence of nitric oxide generation on the production of interleukins and processes of prostaglandin synthesis in fetal membranes in spontaneous parturition and uterine inertia]. AB - 62 women in 38-40 weeks gestation were examined, 37 of them had normal delivery, whereas others (25) with uterine inertia, had delivery by Cesarean section. In the fetal membranes preparations nitric oxide production, interleukins -1beta and -6, arachidonic acid content and phospholipase A2 activity (limiting the rate of prostaglandin biosynthesis) were examined. In the case of uterine inertia increase of nitric oxide generation, decrease of interleukins and arachidonic acid levels and phospholipase A2 activity was found. Modulation of nitric oxide production with inhibitors and activators allowed to emphasize the role of NO in regulation of cytokine response and control of initial stages of prostaglandins synthesis. PMID- 15354543 TI - [Age changes in glycosaminoglycan level in the woman skin]. AB - Postmortem skin samples of 50 women aged from 15 to 90 years were analysed. Protein and DNA levels were stable rather age-independent. Glycosaminoglycane (GAG) level decreased during ageing accordingly linear regression equation: Age = 100.1562 - 0.3578xGAG. PMID- 15354544 TI - [Dual monoaminoxidase and acetylcholinesterase inhibitors as potential agents for treatment of Alzheimer disease]. PMID- 15354548 TI - Sun, Genesis find their footing. Two different approaches, two successful turnarounds. PMID- 15354549 TI - Preserving patient dignity when surrogates step in. PMID- 15354550 TI - MS presents special challenges. PMID- 15354551 TI - When 'strict liability' is abused. PMID- 15354552 TI - Monolithic separation media: where are they heading? PMID- 15354553 TI - Preparation and HPLC applications of rigid macroporous organic polymer monoliths. AB - Rigid porous polymer monoliths are a new class of materials that emerged in the early 1990s. These monolithic materials are typically prepared using a simple molding process carried out within the confines of a closed mold. For example, polymerization of a mixture comprising monomers, free-radical initiator, and porogenic solvent affords macroporous materials with large through-pores that enable applications in a rapid flow-through mode. The versatility of the preparation technique is demonstrated by its use with hydrophobic, hydrophilic, ionizable, and zwitterionic monomers. Several system variables can be used to control the porous properties of the monolith over a broad range and to mediate the hydrodynamic properties of the monolithic devices. A variety of methods such as direct copolymerization of functional monomers, chemical modification of reactive groups, and grafting of pore surface with selected polymer chains is available for the control of surface chemistry. Since all the mobile phase must flow through the monolith, the convection considerably accelerates mass transport within the molded material, and the monolithic devices perform well, even at very high flow rates. The applications of polymeric monolithic materials are demonstrated mostly on the separations in the HPLC mode, although CEC, gas chromatography, enzyme immobilization, molecular recognition, advanced detection systems, and microfluidic devices are also mentioned. PMID- 15354554 TI - Monoliths for fast bioseparation and bioconversion and their applications in biotechnology. AB - Monoliths have consolidated their position in bioseparation. More than 200 different applications have been reported in the past two decades and their advantages compared to conventional chromatography demonstrated. These include the high mass transfer efficiency due to the convective flow enabled by the macroporous character of the matrix. Recently plasmid DNA and viruses were separated with high efficiency and cryogels and monolithic superporous agarose were developed for capture of proteins from crude homogenates and separation of microorganisms or lymphocytes. Currently four companies manufacture monoliths mainly for analytical applications although monoliths with a volume of 0.8 liter are commercially available and 8 L are available as prototypes. A book entitled "Monolithic materials: preparation, properties and applications" was published in 2003 and became standard reference of the status of this area. This review focuses on the progress in monoliths that goes beyond the scope of this reference book. Less progress has been made in the field of bioconversions in spite of the fact that monolithic supports exhibit better performance than beads in enzymatic processing of macromolecules. It appears that the scientific community has not yet realized that supports for these applications are readily available. In addition, monoliths will further substantially advance bioseparations of both small and large molecules in the future. PMID- 15354555 TI - Monolithic columns with a gradient of functionalities prepared via photoinitiated grafting for separations using capillary electrochromatography. AB - Stationary phases for capillary electrochromatography with a longitudinal gradient of functionalities have been prepared via photoinitiated grafting of polymer chains onto the pore surface of a porous polymer monolith. In order to achieve the desired retention and electroosmotic flow, the hydrophobic poly(butyl methacrylate-co-ethylene dimethacrylate) monolith with optimized porous properties was grafted with a layer of ionizable polymer, poly(2-acrylamido-2 methyl-1-propanesulfonic acid). A moving shutter and a neutral density filter were used to control the dose of UV light received at different locations along the monolith in order to create the longitudinal gradient of functionalities. Formation of the desired gradients was confirmed using electron probe microanalysis of different locations along the column. The preparation technique significantly affects performance in the CEC mode as demonstrated on the separations of a model mixture using columns both with homogeneous distribution of grafts and with a gradient of functionality. Columns grafted with the gradient of functionalities were found superior to those functionalized uniformly. A comparison of the performance of the gradient column with another containing evenly distributed functionalities showed the performance benefits of the "gradient" column. PMID- 15354557 TI - Characterization of methacrylate monoliths for purification of DNA molecules. AB - The suitability of methacrylate based anion exchange monolithic supports for the separation and purification of plasmid and genomic DNA has been explored. The effect of the size of the channels, ionic strength of the solution, and ligand density on the dynamic binding capacity has been investigated. The dynamic binding capacity was found to be flow independent, at least up to a linear velocity of 700 cm h(-1), and exceeded 9 mg mL(-1) for all types of DNA. The recovery depends on the pH value of the mobile phase and its ionic strength as well as on the density of the active groups. Under optimal conditions recoveries exceeding 80% were obtained even for genomic DNA. Finally, the suitability of this approach is demonstrated by purification of a real-life sample. PMID- 15354556 TI - Comparison of monolithic silica and polymethacrylate capillary columns for LC. AB - Organic polymer monolithic capillary columns were prepared in fused-silica capillaries by radical co-polymerization of ethylene dimethacrylate and butyl methacrylate monomers with azobisisobutyronitrile as initiator of the polymerization reaction in the presence of various amounts of porogenic solvent mixtures and different concentration ratios of monomers and 1-propanol, 1,4 butanediol, and water. The chromatographic properties of the organic polymer monolithic columns were compared with those of commercial silica-based particulate and monolithic capillary and analytical HPLC columns. The tests included the determination of H-u curves, column permeabilities, pore distribution by inversed-SEC measurements, methylene and polar selectivities, and polar interactions with naphthalenesulphonic acid test samples. Organic polymer monolithic capillary columns show similar retention behaviour to chemically bonded alkyl silica columns for compounds with different polarities characterized by interaction indices, Ix, but have lower methylene selectivities and do not show polar interactions with sulphonic acids. The commercial capillary and analytical silica gel-based monolithic columns showed similar selectivities and provided symmetrical peaks, indicating no significant surface heterogeneities. To allow accurate characterization of the properties of capillary monolithic columns, the experimental data should be corrected for extra-column contributions. With 0.3 mm ID capillary columns, corrections for extra-column volume contributions are sufficient, but to obtain true information on the efficiency of 0.1 mm ID capillary columns, the experimental bandwidths should be corrected for extra-column contributions to peak broadening. PMID- 15354558 TI - Enzyme immobilization on epoxy- and 1,1'-carbonyldiimidazole-activated methacrylate-based monoliths. AB - Monolithic Convective Interaction Media (CIM) have been activated with epoxide and imidazole carbamate functionalities and used as supports for covalent immobilization of protein A, deoxyribonuclease I, and trypsin. The efficiency of immobilization for these proteins was determined from the amount of bound IgG, degradation of DNA, and hydrolysis of Nalpha-benzoyl-L-arginine ethyl ester, respectively. The respective biological activities of trypsin and the binding capacity of protein A immobilized via imidazole carbamate groups were 11.45 and 2.25 times higher than those obtained for epoxide matrix while they were practically equal for deoxyribonuclease I. The kinetics of immobilization was studied in detail for trypsin under dynamic conditions and revealed that the enzyme immobilized via imidazole carbamate groups already reached its highest activity in 5 min. In contrast, a much longer time was required for immobilization via epoxy groups. PMID- 15354559 TI - Mass transfer characteristics of plasmids in monoliths. AB - The hydrodynamic properties and pore-structure of monoliths based on functionalized poly(glycidyl methacrylate-ethylene dimethacrylate) were characterised by pulse response experiments using different probes representing a wide range of molecular mass. On a small scale, band spreading was found to be caused to the extent of more than 90% by extra-column effects. These monoliths have large channel diameters, providing a suitable chromatography adsorbent for processing of large molecules. Dynamic and static binding capacity for plasmid DNA was investigated. For our model plasmid, consisting of 4.9 kbp, a capacity of 7 mg/mL was observed in comparison to 0.3 mg/mL for a conventional medium designed for protein separation. When plasmids were loaded on the monolith a gradual increase in pressure drop was observed. The channels filled up and the cross-sectional area available for liquid flow decreased. Therefore, a higher pressure drop was observed during elution. This is caused by (i) shrinking of the channels as effect of the high salt concentration, (ii) high viscosity of the mobile phase due to high concentration of plasmids, and (iii) an increase of the hydrodynamic radius of the plasmid with salt concentration from 45 nm at 150 mM to 70 nm at 2 M NaCl, as measured by dynamic light scattering. These types of monoliths are considered to be the preferred adsorbents for plasmid separation. PMID- 15354560 TI - Characterization of polyacrylamide based monolithic columns. AB - Supermacroporous monolithic polyacrylamide (pAAm)-based columns have been prepared by radical cryo-copolymerization (copolymerization in the moderately frozen system) of acrylamide with functional co-monomer, allyl glycidyl ether (AGE), and cross-linker N,N'-methylene-bis-acrylamide (MBAAm) directly in glass columns (ID 10 mm). The monolithic columns have uniform supermacroporous sponge like structure with interconnected supermacropores of pore size 5-100 microm. The monoliths can be dried and stored in the dry state. High mechanical stability of the monoliths allowed sterilization by autoclaving. Column-to-column reproducibility of pAAm-monoliths was demonstrated on 5 monolithic columns from different batches prepared under the same cryostructuration conditions. PMID- 15354561 TI - Molecularly imprinted polymers: a new tool for separation of steroid isomers. AB - Molecularly imprinted polymers hold great promise for the separation of chiral compounds. A non-covalent approach to the synthesis of MIPs relies on the presence of specific and non-specific interactions, which lead to the formation of a molecular imprint. The structural differences between 17-alpha-estradiol and 17-beta-estradiol are too small to permit their efficient separation on a MIP prepared with beta-estradiol as a template. Molecular modeling revealed the presence of only one hydrogen bond that differentiates the two isomers. PMID- 15354562 TI - Applications of silica-based monolithic HPLC columns. AB - The recent invention and successive commercial introduction of monolithic silica columns has motivated many scientists from both academia and industry to study their use in HPLC. The first paper on monolithic silica columns appeared in 1996. Currently about 200 papers have been published relating to applications and characterization of monolithic silica columns, including monolithic capillaries. This review attempts to give an overview covering various aspects of this new column type in the field of high throughput analysis of drugs and metabolites, chiral separations, analysis of pollutants and food-relevant compounds, as well as in bioanalytical separations such as in proteomics. Some of the applications are described in greater detail. The numerous publications dealing with the physicochemical and chromatographic characterization of monolithic silica columns are briefly summarized. PMID- 15354563 TI - Characterization of monolithic columns for HPLC. AB - Monolithic stationary phases and columns have rapidly become highly popular separation media for liquid chromatography, in spite of their recent discovery. However, their most important features have not yet been completely clarified. A complete understanding of their performance and of their intrinsic characteristics will require the systematic acquisition of many series of reliable experimental data and their consistent analysis from different points of view. Progress in their design and production requires now that the chromatographic behavior of monolithic columns be studied in close connection with their physico-chemical and structural properties. The main goal of this review is to summarize fundamental information on some physico-chemical and chromatographic characteristics of monolithic stationary phases and columns for RPLC. The material reviewed deals only with silica-based monolithic columns. First, structural information on the porosities and the size of the pores in monolithic columns is reported. Second, results of chromatographic studies that deal with the characterization of monolithic columns are summarized. Third, results of detailed studies made on the adsorption equilibrium and the surface heterogeneity of monolithic stationary phases are presented. Finally, results on the mass transfer kinetics in monolithic columns derived from the applications of the classical random-walk model and of the moment theory to a new model of the monolith are discussed. PMID- 15354565 TI - A computational study of the porosity effects in silica monolithic columns. AB - We report on a theoretical study of the influence of the through-pore porosity on the main chromatographic performance parameters (reduced theoretical plate height, flow resistance, and separation impedance) of silica monoliths. To investigate this problem devoid of any structural uncertainties, computer generated structural mimics of the pore geometry of silica monolithic columns have been studied. The band broadening in these synthetic monoliths was determined using a commercial Computational Fluid Dynamics (CFD) software package. Three widely differing external porosities (epsilon = 0.38, epsilon = 0.60, and epsilon = 0.86) are considered and are compared on the basis of an identical intra-skeleton diffusivity (Ds = 5 x 10(-10)m2/s), internal porosity (epsilon(int) = 0.5), and for the same phase retention factor (k' = 1.25). Since the data are obtained for perfectly ordered structures, the calculated plate heights and separation impedances constitute the ultimate performance ever to be expected from a monolithic column. It is found that, if silica monoliths could be made perfectly homogeneous, domain size-based reduced plate heights as small as h(min) approximately 0.8 (roughly independent of the porosity) and separation impedances as small as Emin approximately 130 (epsilon = 0.60) and Emin approximately 40 (epsilon = 0.86) should be achievable with pure water as the working fluid. The data also show that, although the domain size is a much better reduction basis than the skeleton size, the former is still not capable of bringing the van Deemter curves of different porosity columns into perfect agreement in the C term dominated velocity range. It is found that, in this range, large porosity monoliths can be expected to yield smaller domain size based reduced plate heights than small porosity monoliths. PMID- 15354564 TI - Structural formation of hybrid siloxane-based polymer monolith in confined spaces. AB - Structural deformation of phase-separated methylsiloxane gel under the influence of a surface has been studied. Competitive wetting of siloxane gel phase on a surface during phase formation is found to significantly affect the final morphology in a confined space. When the spinodal wavelength is sufficiently shorter than the size of the available space, a uniform bicontinuous structure forms in confined geometry. However, gel skeletons in the vicinity of a surface are elongated with decreasing size of the space, and finally when the size of the space becomes shorter than the spinodal wavelength, all the gel phase wets on a surface, showing a "wetting transition". Homogeneous bicontinuous methylsiloxane gels were successfully prepared, avoiding such structural deformation, in a long cylindrical fused silica capillary and used for capillary HPLC. The capillary gels exhibited excellent separation efficiency of nitrobenzenes and it was found that the surface character can be altered by incorporating surfactants, which will enable more advanced and extended control of surface character, depending on the analytes. PMID- 15354567 TI - High-performance liquid chromatographic enantioseparations on capillary columns containing monolithic silica modified with cellulose tris(3,5 dimethylphenylcarbamate). AB - Monolithic capillary columns containing native silica gel were modified with cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) and used for enantioseparations in capillary liquid chromatography. The method adopted for in situ enantioselective modification of monolithic fused silica capillary columns by coating with CDPMC appears to be fairly simple and fast. High efficiency enantioseparations of test racemic compounds and s(everal chiral drugs were achieved in a short time. It was possible to increase the amount of chiral selector present by multiple coating of monoliths with CDMPC. The baseline enantioseparation of 2,2,2-trifluoro-1-(9-anthryl)ethanol was achieved in an analysis time less than 30 s with this capillary column. In addition, reproducible enantioseparations were obtained when the chiral selector was removed from the monolithic column by flushing it with appropriate solvent and the column recoated. PMID- 15354566 TI - Simple 2D-HPLC using a monolithic silica column for peptide separation. AB - Separation of peptides by fast and simple two-dimensional (2D)-HPLC was studied using a monolithic silica column as a second-dimension (2nd-D) column. Every fraction from the first column, 5 cm long (2.1 mm ID) packed with polymer-based cation exchange beads, was subjected to separation in the 2nd-D using an octadecylsilylated (C18) monolithic sillica column (4.6 mm ID, 2.5 cm). A capillary-type monolithic silica C18column (0.1 mm ID, 10 cm) was also employed as a 2nd-D column with split flow/injection. Effluentof the first dimension (1st D) was directly loaded into an injector loop of 2nd-D HPLC. UV and MS detection were successfully carried out at high linear velocity of mobile phase at 2nd-D using flow splitting for the 4.6 mm ID 2nd-D column, or with directconnection of the capillary column to the MS interface. Two-minute fractionation inthe 1st-D, 118-second loading, and 2-second injection by the 2nd-D injector, allowed one minute for gradient separation in the 2nd-D, resulting in a maximum peak capacity of about 700 within 40 min. The use of a capillary column in solvent consumption and better MS detectability compared to a larger-sized column. This kind of fast and simple 2D-HPLC utilizing monolithic silica columns will be useful for the separation of complex mixtures in a short time. PMID- 15354568 TI - Rapid, low pressure, and simultaneous ion chromatography of common inorganic anions and cations on short permanently coated monolithic columns. AB - Short permanently coated reversed-phase silica based monolithic columns have been investigated for the rapid separation of inorganic anions and cations. One 2.5 x 0.46 cm column was permanently coated with didodecyldimethylammonium (DDAB), for anion analysis; and a second 5.0 x 0.46 cm column was coated with dioctylsulphosuccinnate (DOSS), for cation analysis. The use of a single combined eluent of 2.5 mM phthalate/1.5 mM ethylenediamine, at flow rates of between 4.0 and 8.0 mL/min, resulted in the rapid separation of 8 anions (in under 100 s) and 5 cations (in under 100 s) on the above columns when used individually, with detection limits for common anions ranging from approximately 0.25 to 5 mg/L, and between 2.5 and 50 mg/L for alkaline earth metals, by direct and indirect conductivity detection, respectively. However, with both columns subsequently connected in parallel, with the eluent delivered using a flow splitter from a single isocratic pump, the simultaneous analysis of anions and cations was also possible, based on a single conductivity detector. The potential of this system for the rapid, complete screening of water samples for multiple common anions and cations is shown. PMID- 15354569 TI - Ion exchange properties of monolithic and particle type iminodiacetic acid modified silica. AB - A 10 cm silica monolith has been modified with iminodiacetic acid (IDA) groups and characterised for its selectivity toward alkali, alkaline earth, and selected transition metal cations. Physical characterisation of the modified monolith found non-homogeneous modification along the length of the monolith, although sufficient capacity was achieved to facilitate significant retention of alkaline earth and transition/heavy metal ions over a range of eluent pH and ionic strength conditions. For alkaline earth and transition/heavy metal ions, selectivity of the 10 cm IDA monolith closely matched that seen with a 25 cm IDA modified silica gel particle packed column, although the separation of alkali metal ions was noticeably poorer on the monolithic column. Peak efficiencies for most metal ions were of a similar order for both column types, except for Zn(II), which showed significant peak broadening on the IDA monolithic column. PMID- 15354570 TI - Strong phylogeographic relationships among three goat breeds from the Canary Islands. AB - We partially sequenced the mitochondrial D-loop region in 47 individuals from eleven Spanish and foreign goat breeds. Phylogenetic analysis of these sequences allowed us to identify a particular D-loop haplotype shared by individuals from the Palmera, Majorera and Tinerfena Canarian breeds. Genotyping of 281 goats from 17 different breeds by PCR-Hpall RFLP evidenced that the geographical distribution of this haplotype is restricted to the Canary Islands. This ancestral mitochondrial haplotype might originate in the domestic goat herds brought by the native Canarian inhabitants approximately 3000 years ago. Although we observed other miscellaneous D-loop haplotypes in the Palmera, Majorera and Tinerfena breeds, any of them allowed us to group individuals from these three populations in a single cluster, a feature that suggests that these haplotypes might have diverse origins. The remarkable degree of phylogeographic structure of the Canary goat breeds with regard to other Spanish and foreign populations might be attributed to the isolation of these breeds in the Canary Islands for approximately 2500 years, without exposure to the migratory movements and commercial trading events that probably affected the genesis of most domestic goat breeds worldwide. The Canarian D-loop haplotype can be efficiently genotyped by using DNA isolated from milk and cheese samples, which paves the way for the future establishment of a Canary breed identity test for these dairy products. PMID- 15354571 TI - Physical properties of mammary secretions in relation to chemical changes during transition from colostrum to milk. AB - We examined the physical and chemical changes in milk during early lactation, and how these changes were affected by leaving one quarter unmilked in either the first or second milking, with the purpose of discriminating between colostrum and normal milk. Milk samples were collected from each quarter of 17 cows during the first 5 d after calving and then after about 7 d and 14d. Samples were analysed for somatic cell count (SCC), fat, protein, casein, lactose, IgG1, colour, plasmin, pH and coagulation properties. Large variations occurred in both chemical and physical properties throughout the study period. Within six milkings, the concentration of casein decreased by 60%, IgG1 by 94%, and lactose increased by 34%. At milking number 6, rennet coagulation time was lowest and curd firmness was highest. The pH increased from 6.4 to 6.7 over the period of the experiment, and the colour changed from yellow (reddish) to white. Coagulation properties and the pH fell within the range of normal milk after five milkings. Measurement of colour and density appeared to be a potential method for detection of milk unsuitable for the dairy factory. Effects of omitting one quarter in one milking differed between milk components, but seemed to be of little importance to the physical properties. PMID- 15354572 TI - Effect of vitamin E supplementation on neutrophil function, milk composition and plasmin activity in dairy cows in a commercial herd. AB - Fifty-six Holstein dairy cows from a commercial dairy herd in the Northern part of Greece were used to determine the effect of vitamin E supplementation on immune parameters, milk composition and milk quality. Cows were assigned to one of two experimental groups: control (no vitamin E supplementation) and vitamin E supplementation. Supplementation of vitamin E started 4 weeks prior to and continued up to 12 weeks after parturition. Supplementation included daily oral administration of vitamin E at 3000 i.u./cow prepartum and was reduced to 1000 i.u./cow post partum. Blood samples were collected weekly for 8 weeks starting 4 weeks before parturition, neutrophils were isolated and the following parameters were determined in neutrophils activated by phorbol myristate acetate: total cell associated and membrane-bound urokinase plasminogen activator (u-PA) activity and superoxide production. Milk samples were collected weekly and fat, protein, lactose, somatic cell count (SCC), plasmin and plasminogen-derived activity were determined. Activated neutrophils isolated from cows that received supplemental vitamin E had higher (P<0.01) total and membrane-bound u-PA activities during the first 3 weeks after parturition and higher (P<0.01) superoxide production during week 1 prepartum and week 1 post partum compared with the corresponding values of activated neutrophils isolated from control cows. Vitamin E supplementation had no effect (P=0.28) on plasminogen-derived activity in milk. Milk obtained from cows that received supplemental vitamin E had SCC lower by 25% (P<0.05) and plasmin lower by 30% (P<0.01) than corresponding values in milk obtained from control cows. The reduction in plasmin as a result of vitamin E supplementation is very beneficial to the dairy industry because plasmin reduces the cheese yielding capacity of milk, affects the coagulating properties of milk and its overall ability to withstand processing during cheesemaking. In conclusion, vitamin E supplementation had positive effects on the function of bovine neutrophils and milk quality in a commercial dairy herd. PMID- 15354573 TI - Effects of feeding or abomasal infusion of canola oil in Holstein cows. 1. Nutrient digestion and milk composition. AB - We determined the effects of feeding canola oil or infusing it into the abomasum on rumen fermentation, nutrient digestibility, duodenal flows of fatty acids, and milk composition in Holstein cows. Five ruminally and duodenally cannulated Holstein cows in late lactation were used in a 3 x 5 incomplete Latin square design. Treatments were 1) CONTROL: basal diet (CON), 2) CONTROL+supplementation of canola oil at 1 kg/d in the feed (FED), and 3) CONTROL+abomasal infusion of canola oil at 1 kg/d (INF). Compared with CON, feed intake, ruminal fermentation characteristics, ruminal and total tract digestibilities of nutrients were not significantly affected by FED treatment but duodenal flows and milk concentrations of fatty acids (FA) such as trans-11 18:1 and cis-9 trans-11 18:2 (conjugated linoleic acid, CLA) were increased. In contrast to the effects of FED, INF reduced feed intake, total VFA production, intestinal flows of nutrients, FA digestibility and yields of milk and milk fat. Both FED and INF significantly reduced the proportions of saturated and medium-chain FA, and increased cis 18:1 in milk. Concentrations of 18:2n-6 and 18:3n-3 in milk were increased nearly 2-fold with INF relative to CON. Dietary or postruminal supplementation of canola oil to late-lactation cows reduced saturated FA and increased unsaturated C18 in milk but nutrient digestion was adversely affected with abomasal infusion of canola oil. PMID- 15354574 TI - Effects of feeding or abomasal infusion of canola oil in Holstein cows. 2. Gene expression and plasma concentrations of cholecystokinin and leptin. AB - We determined the relative importance of cholecystokinin (CCK), leptin, and fatty acid concentrations in plasma in mediating the satiety effects of supplemental fat in lactating cows. Five ruminally and duodenally cannulated Holstein cows in late lactation were used in a 3 x 5 incomplete Latin square design with three treatments: 1) CONTROL: basal diet (CON), 2) CONTROL+supplementation of canola oil at 1 kg/d in the feed (FED) and 3) CONTROL+abomasal infusion of canola oil at 1 kg/d (INF). Relative to CON, feed intake was reduced by INF but not by FED. We provide evidence that both FED and INF treatments stimulated CCK gene expression in the duodenum and elevated plasma CCK concentrations. However, our results did not support a role for CCK in mediating satiety through an endocrine mechanism of action. We speculate that CCK might be acting either through paracrine and/or neurocrine routes to influence feed intake in cattle. Both FED and INF had no effect on the mRNA abundance of leptin, lipoprotein lipase, or acetyl-CoA carboxylase in adipose tissue. Plasma concentrations of leptin, insulin and IGF-I were not altered by FED or INF, indicating that these signals may not be involved in mediating short-term hypophagic effects of dietary fat. Plasma concentrations of 18:1n-9 and 18:2n-6 were significantly greater for INF than for FED or CON. We conclude that the hypophagic effects of supplemental fat in cattle depend on the amount of unsaturated fatty acids reaching the intestine and that this satiety effect is mediated through CCK, oleic acid and (or) linoleic acid, but leptin is not involved. PMID- 15354575 TI - Increasing magnesium intakes in relation to magnesium absorption in dry cows. AB - Earlier studies with temporarily isolated rumen of heifers show saturation kinetics of Mg efflux across the rumen wall. Therefore, we hypothesized that high Mg intakes would not further increase the rate of Mg absorption in cows. To test our hypothesis, six ruminally fistulated non-pregnant dry cows were given diets with different Mg concentrations in a 6 x 6 Latin square design. Desired concentrations of Mg were attained by adding MgO to the basal diet and the Mg concentrations in the total rations were 3.8, 6.4, 9.1, 11.8, 14.1 and 173 g Mg/kg dry matter, which provided Mg intakes of 27.1, 44.6, 64.6, 83.5, 100.4 and 124.3 g/d, respectively. Increasing Mg intakes were associated with increased (P<0.001) faecal Mg excretion. However, apparent Mg absorption expressed as g/d was not significantly different for Mg intakes from 100.4 to 124.3 g/d while Mg absorption expressed as a proportion of intake was not significantly different for Mg intakes ranging from 64.6 to 124.3 g/d. Mg concentrations in rumen fluid after feeding increased (P<0.001) with increasing Mg intakes. Apparent absorption of Mg appeared to become saturated at a ruminal Mg concentration of 17.5 mM (Mg intake of 83.5 g/d). Group-mean post-feeding concentrations of Mg and Na in rumen fluid were significantly correlated (Pearson's r=-0.96; P=0.003, n=6). This study showed that under conditions of practical dairy cow feeding, Mg absorption was maximal at Mg intakes > or = 84 g/d. PMID- 15354576 TI - Effect of different milking intervals on the composition of cisternal and alveolar milk in dairy cows. AB - Effects of six different milking intervals on the distribution of milk between cistern and alveoli were studied in a randomized, incomplete Latin Square experiment with four lactating Holstein cows. Cisternal and alveolar milk was measured by udder quarter at 4, 8, 12, 16, 20 and 24-h intervals with a 3-d interperiod of regular milking. Cisternal milk was evacuated using a cannula after injection of an oxytocin-receptor blocking agent, followed by an injection of oxytocin to remove the alveolar fraction. Milk samples from each fraction and quarter were collected for analysis. Cisternal and alveolar milk increased with milking interval and represented on average 30 and 70% of the milk stored in the udder, respectively. Fat content in alveolar milk remained constant during the first 16 h, increasing rapidly thereafter, reaching its maximum at 24 h (6.95%). Fat content in cisternal milk decreased with milking interval and reached its minimum at 24 h (0.96%). Total fat yield tended to increase for cisternal milk with longer milking intervals, but it increased markedly for alveolar milk, showing that fat globules did not pass freely from alveoli to cistern between milkings. Milk protein content was greater in rear quarters than in front quarters for both milk fractions. Milk protein content increased in the cisternal milk fraction and tended to increase in the alveolar milk fraction with longer milking intervals, but values did not differ between cisternal and alveolar fractions or between front and rear quarters. Total protein yield increased with milking interval in both fractions, indicating that casein micelles passed more freely than fat globules from the alveolar to the cisternal compartment. In conclusion, the short-term effects of milking intervals in milk composition were explained by the changes observed in alveolar and cisternal milk ratio. PMID- 15354577 TI - Effect of subclinical intramammary infection on somatic cell counts, NAGase activity and gross composition of goats' milk. AB - The study was aimed at identifying the pathogens causing subclinical udder infections in representative Israeli dairy goat herds and determining their effect on milk quality. Five hundred goats in ten flocks of various breeds and crossbreeds were surveyed. Of the 500 goats, 13.4% were in their first lactation, 36.4% were in their second lactation and 50.2% were in their third or higher lactation. Percentages of udder halves with subclinical intramammary infection in the flocks ranged from 35 to 71%. The effect of the bacteriological infection on somatic cells count (SCC) was significant (P<0.001). Various species of coagulase negative staphylococci (CNS), mainly Staphylococcus caprae and Staphylococcus epidermidis, were the main pathogens in infected udder halves. Lactation number did not significantly influence either infection rate of udder halves or SCC, although the percentage of udder halves with no bacteriological findings was higher at the first lactation than at the third lactation. Milk composition (fat, protein and lactose) varied among flocks, with lower mean total protein in uninfected halves than in infected ones and higher lactose in uninfected than infected halves. PMID- 15354578 TI - Changes of physicochemical indicators during mastitis and the effects of milk ejection on their sensitivity. AB - We examined the relationship between physicochemical indicators and somatic cells in the milk of dairy cows during experimentally induced mastitis and their significance as indicators for use in controlling udder health. We were concerned particularly with the effect of alveolar milk ejection on the sensitivity of these indicators. In Expt 1, Escherichia coli lipopolysaccharide (Esch. coli LPS) was injected into the left rear quarter to induce an inflammatory reaction in one quarter in each of six cows. The contralateral control quarter was injected with a solution of NaCl (9 g/l). Nine milk samples were taken from both quarters until 60 h after injection. In Expt 2, repeated milk samples were taken every 20 s from one quarter during a 120-s teat stimulation in 20 cows with different somatic cell counts (SCC). Quarters were clustered for low (<5.0 log cells/ml), mid (5.0 5.7 log cells/ml) and high (>5.7 log cells/ml) SCC of the sample taken at t=0 s. Samples were analysed for SCC, electrical conductivity (EC) and Na+ and Cl- concentrations. During the experimental inflammation SCC, EC, Na+ and Cl- peaked at 12 h from LPS administration and values in treated quarters (T) at this time were elevated to 7900, 157, 501 and 169% of the values in untreated quarters, respectively. In Expt 2, SCC, EC, Na+ and Cl- in high SCC quarters were 2520, 121, 283 and 141% of low SCC quarters at the start of stimulation (t=0 s), respectively. Highly significant (P<0.001) differences in EC, Na+ and Cl- between high and low SCC quarters disappeared owing to the onset of alveolar milk ejection 100 s after the first contact with the teat. In conclusion, SCC in cows' milk provided the strongest amplitude in the case of an intramammary inflammation. EC, Na+ or Cl- were useful tools only if the measurements were performed in cisternal milk before the start of alveolar milk ejection. PMID- 15354579 TI - Fast biosensor immunoassays for the detection of cows' milk in the milk of ewes and goats. AB - Two monoclonal antibodies (MAb) raised against bovine kappa-casein were developed and applied in an automated optical biosensor (Biacore 3000) to create easy and fast direct and inhibition biosensor immunoassays (BIA) for the detection of cows' milk in the milk of ewes and goats. With both assay formats, low limits of detection (<01%) and fast run times (around 5 min) were obtained. For sample preparation, milk was diluted in buffer (direct assay) or in an antibody containing buffer (inhibition assay) only. For quantitative analysis, calibrants of cows' milk in ewes' or goats' milk were used. Advantages of the direct BIA are: the single reagent format (biosensor chip immobilized antibodies only); the use of small amounts of antibodies (2 microg for >350 tests); and the wide measurement range (0.1 to 10% cows' milk). Despite these advantages, the inhibition BIA (using kappa-casein immobilized on the chip) was preferred because of the possible application of non-purified Mab, the higher responses, the higher sensitivity at relevant low percentages of cows' milk and its robustness (>800 cycles per chip). PMID- 15354580 TI - The roles of disulphide and non-covalent bonding in the functional properties of heat-induced whey protein gels. AB - Heat-induced gelation (80 degrees C, 30 min or 85 degrees C, 60 min) of whey protein concentrate (WPC) solutions was studied using transmission electron microscopy (TEM), dynamic rheology and polyacrylamide gel electrophoresis (PAGE). The WPC solutions (150 g/kg, pH 6.9) were prepared by dispersing WPC powder in water (control), 10 g/kg sodium dodecyl sulphate (SDS) solution or 10 mM dithiothreitol (DTT) solution. The WPC gels containing SDS were more translucent than the control gels, which were slightly more translucent than the gels containing DTT. TEM analyses showed that the SDS-gels had finer aggregate structure (approximately equal to 10 nm) than the control gels (approximately equal to 100 nm), whereas the DTT-gels had a more particulate structure (approximately equal to 200 to 300 nm). Dynamic rheology measurements showed that the control WPC gels had storage modulus (G) values (approximately equal to 13,500 Pa) that were approximately equal to 25 times higher than those of the SDS gels (approximately equal to 550 Pa) and less than half those of the DTT-gels after cooling. Compression tests showed that the DTT-gels were more rigid and more brittle than the control gels, whereas the SDS-gels were softer and more rubbery than either the control gels or the DTT-gels. PAGE analyses of WPC gel samples revealed that the control WPC solutions heated at 85 degrees C for 10 min contained both disulphide bonds and non-covalent linkages. In both the SDS solutions and the DTT-solutions, the denatured whey protein molecules were in the form of monomers or small aggregates. It is likely that, on more extended heating, more disulphide linkages were formed in the SDS-gels whereas more hydrophobic aggregates were formed in the DTT-gels. These results demonstrate that the properties of heat-induced WPC gels are strongly influenced by non covalent bonding. Intermolecular disulphide bonds appeared to give the rubbery nature of heat-induced WPC gels whereas non-covalent bonds their rigidity and brittle texture. PMID- 15354581 TI - Selection of acid tolerant bifidobacteria and evidence for a low-pH-inducible acid tolerance response in Bifidobacterium longum. AB - Acidity is an environmental condition commonly encountered by lactic acid bacteria and bifidobacteria in the gastrointestinal tract and fermented foods. In the present study, 22 strains of Bifidobacterium were screened for acid tolerance in artificial gastric juice (AGJ, pH 3.0) and fermented milk. AGJ tolerance was found to be strain-specific, with a pronounced variation among the strains. Several strains with a high survival rate in AGJ that belonged to Bifid. longum, Bifid. breve and Bifid. adolescentis were selected. Among them, only strain BL1 of Bifid longum was found to possess a high survival rate in fermented milk during refrigerated storage. Strain BL1 exhibited a survival rate of more than 25% in AGJ at pH 3.0 for 2 h and maintained a viable cfu level of more than 10(8) per gram of product in fermented milk (pH 4.6) under refrigerated conditions for 2 weeks. The acid tolerance of strain BL1 was found to depend on the final growth pH (<4.5). Rapid loss of acid tolerance was observed when the cells were shifted from acid to neutral conditions by addition of NaOH. Strain BL1 cells were able to maintain much higher intracellular pH under acid conditions, in comparison with those of AGJ sensitive mutant (BL1-S) or cells that lost acid tolerance following pH shifting from acid to neutral conditions. These results suggested that a cytoplasmic pH homeostasis system may function in the acid tolerance response in this strain. PMID- 15354582 TI - Controlled production of Camembert-type cheeses. Part I: Microbiological and physicochemical evolutions. AB - A holistic approach of a mould cheese ripening is presented. The objective was to establish relationships between the different microbiological and biochemical changes during cheese ripening. Model cheeses were prepared from pasteurized milk inoculated with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti and Brevibacterium linens under aseptic conditions. Two cheese-making trials with efficient control of environmental parameters were carried out and showed similar ripening characteristics. K. lactis grew rapidly between days 1 and 6 (generation time around 48 h). G. candidum grew exponentially between days 4 and 10 (generation time around 4.6 d). Brevi. linens also grew exponentially but after day 6 when Pen. camemberti mycelium began developing and the pH of the rind was close to 7. Its exponential growth presented 3 phases in relation to carbon and nitrogen substrate availability. Concentrations of Pen. camemberti mycelium were not followed by viable cell count but they were evaluated visually. The viable microorganism concentrations were well correlated with the carbon substrate concentrations in the core and in the rind. The lactose concentrations were negligible after 10 d ripening, and changes in lactate quantities were correlated with fungi flora. The pH of the inner part depended on NH3. Surface pH was significantly related to NH3 concentration and to fungi growth. The acid-soluble nitrogen (ASN) and non-protein nitrogen (NPN) indexes and NH3 concentrations of the rind were low until day 6, and then increased rapidly to follow the fungi concentrations until day 45. The ASN and NPN indexes and NH3 concentrations in the core were lower than in the rind and they showed the same evolution. G. candidum and Pen. camemberti populations have a major effect on proteolysis; nevertheless, K. lactis and Brevi. linens cell lysis also had an impact on proteolysis. Viable cell counts of K. lactis, G. candidum, Pen. camemberti and Brevi. linens were correlated with the environmental conditions, with proteolytic products and with carbon substrate assimilation. NH3 diffusion from surface to the cheese core during ripening was highly suspected. Interaction phenomena between microorganisms are discussed. PMID- 15354583 TI - Controlled production of Camembert-type cheeses. Part II. Changes in the concentration of the more volatile compounds. AB - Flavour generation in cheese is a major aspect of ripening. In order to enhance aromatic qualities it is necessary to better understand the chemical and microbiological changes. Experimental Camembert-type cheeses were prepared in duplicate from pasteurized milk inoculated with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti and Brevibacterium linens under aseptic conditions. Two replicates performed under controlled conditions of temperature (12 degrees C), relative humidity (95 +/- 2%), and atmosphere showed similar ripening characteristics. The evolutions of metabolite concentrations were studied during ripening. The volatile components were extracted by dynamic headspace extraction, separated and quantified by gas chromatography and identified by mass spectrometry. For each cheese the volatile concentrations varied with the part considered (rind or core). Except for ethyl acetate and 2 pentanone, the volatile quantities observed were higher than their perception thresholds. The flavour component production was best correlated with the starter strains. During the first 10 days the ester formations (ethyl, butyl and isoamyl acetates) were associated with the concentrations of K. lactis and G. candidum. The rind quantity of esters was lower than that observed in core probably due to (1) a diffusion from the core to the surface and (2) evaporation from the surface to the chamber atmosphere. G. candidum and Brev. linens association produced 3 methyl butanol and methyl 3-butanal from leucine, respectively. DMDS came from the methionine catabolism due to Brev. linens. Styrene production was attributed to Pen. camemberti. 2-Pentanone evolution was associated with Pen. camemberti spores and G. candidum. 2-Heptanone changes were not directly related to flora activities while 2-octanone production was essentially due to G. candidum. This study also demonstrates the determining role of volatile component diffusion. PMID- 15354584 TI - Influence of manufacturing conditions on the conjugated linoleic acid content and the isomer composition in ripened French Emmental cheese. AB - In a study of the evolution of conjugated linoleic acid (CLA) during cheese production, the influence of Emmental cheese processing on the CLA content and the CLA isomer composition was evaluated. The use of raw and thermised milk, changes of processing temperature and the effect of propionic acid bacteria (PAB) were investigated. The content of CLA in raw milk was 8.6 +/- 1.9 mg/g fat and in the ripened cheese at 70 d was 8.6 +/- 1.6 mg/g fat, under normal processing conditions. No changes in the CLA content and CLA isomer composition were observed during Emmental cheese manufacturing process. Changes in cooking and moulding temperatures did not influence the CLA content. CLA content of cheese made from microfiltered milk with two different Propionibacterium freudenreichii strains was very close to cheeses made without PAB. CLA levels seem to be stable in this type of dairy product under the conditions examined. PMID- 15354585 TI - Changes in lipid fractions and sensory properties of Idiazabal cheese induced by lipase addition. AB - This work studied the addition of an adequate lipase to enhance lipolysis reactions and the development of piquant flavour and sharp odour in Idiazabal cheese, as an alternative to the use of lamb rennet paste. Cheeses were manufactured from bulk raw ewes' milk in 50 l vats with commercial bovine rennet and 80 lipase units of pregastric or 180 lipase units of fungal lipase and ripened for 180 days. A higher lipolytic activity was induced by lipase addition promoting strong changes in odour and flavour attributes. Both fungal and pregastric lipases increased the content of total free fatty acids (FFA), but the fungal lipase released mainly medium- and long-chain FFA. In contrast, the pregastric lipase preferably released short-chain FFA. Diglyceride (DG) content was considerably higher in cheeses made with added pregastric lipase compared with those made with fungal lipase or with no lipase. Monoglycerides (MG) were detected only in cheeses made with either lipase added, reaching comparable concentrations after ripening for 180 days. The cheeses made with pregastric lipase had the highest scores for odour and flavour intensity, and sharp and rennet odours, desirable attributes for the Idiazabal cheese made with lamb rennet paste. None of the texture attributes were significantly influenced by the concentrations of MG and DG in the cheeses made with either lipase. Thus, the pregastric lipase was more appropriate than the fungal lipase to develop a more traditionally-flavoured Idiazabal cheese. PMID- 15354586 TI - Volatile compounds in cheeses made from raw ewes' milk ripened with a lactic culture. PMID- 15354587 TI - Inpatient heart failure treatment from the patient's perspective. AB - OBJECTIVE: The objective of this study was 2-fold: (1) to identify particular opportunities for improvement in patient-centered care of heart failure patients and (2) to suggest strategies for service quality improvement focusing on those areas. SAMPLE: A national cross-sectional sample of survey data from diagnostic related group 127 patients was collected between December 1, 2001, and November 30, 2003. Data were split into two 12-month samples to compare results over time. The 2002 sample included 5224 patients treated at 220 hospitals; the 2003 sample included 6531 patients treated at 269 hospitals. METHOD: A standardized mail out/mail-back methodology was used to collect data from random samples of patients within 5 days of discharge. RESULTS: For both samples, the ranking of service issues was highly similar, with the same 4 areas emerging as the foremost priorities: patient involvement in decision making, staff response to concerns voiced during the hospital stay, staff sensitivity to the inconvenience of heart failure and hospitalization, and emotional/spiritual support. Improvement in these 4 service areas should be associated with the greatest increases in patient satisfaction and quality of care for heart failure patients. CONCLUSIONS: Adequately addressing these patient needs should increase patient satisfaction and quality of care for heart failure patients. PMID- 15354588 TI - Influence of key variables on the patients' choice of a physician. AB - BACKGROUND: Customer orientation is becoming as important in providing health care services as it is in other services. In addition to expecting good health care, patients are also expecting good customer service from their health care providers. Selection of a family physician is a deliberate choice process. OBJECTIVE: This study investigated the variables that influence the choice of a family care physician. METHOD: This study used an experimental design to investigate the influence of physician's communication style, physician's expertise and role of office staff, appointment availability, etc on the selection of a physician. RESULTS: The results show the significance of physician's expertise and the role of office staff in the choice of a physician. PMID- 15354589 TI - Improving hypertension care with patient-generated run charts: physician, patient, and management perspectives. PMID- 15354590 TI - Appropriateness of care: the case for changing the focus of "quality" measurement. AB - "Quality" of care has become one of the main foci of the American health care system during the last 20 years. As costs escalated in all areas of health care, the purchasers and consumers of health care have begun to demand accountability for the products they purchase. The initial focus of the system was "quality of care." When a number of research studies and reports highlighted the lack of basic safety in the American health care system, "safety" of health care was added to the discussion. It is the intent of this article to argue that quality of care is at this point in time not measurable, but that "appropriateness" of care is and can be measured based upon knowledge-based guidelines. With the option to use knowledge-based guidelines as sources for appropriateness indicators, the opportunity to add meaning to indicators and their meaning to the many stakeholders interested in this area cannot be ignored. PMID- 15354591 TI - The theory of constraints and medical error: a conversation with Robert A. McNutt. PMID- 15354592 TI - Reducing radiocontrast use in the cardiac catheterization laboratory--a behavioral modification approach. AB - OBJECTIVES: This study assessed the impact of behavioral modification techniques on reducing the volume of contrast dye used during cardiac catheterization and percutaneous coronary intervention (PCI). BACKGROUND: Excessive administration of radiocontrast agents in the catheterization laboratory is associated with numerous adverse effects including radiocontrast nephropathy. This may be precipitated by using large volumes of contrast dye, particularly in diabetic patients and those with pre-existing renal dysfunction. METHODS: Data were prospectively collected on 20,322 consecutive patients undergoing cardiac catheterization and PCI between January 1997 and December 2001. A focused behavioral modification program was initiated in late 1998. The physician was informed verbally during the course of the procedure and by letter after the procedure regarding the volume of contrast dye used for cases in which the radiocontrast volume exceeded 300 mL. Data were analyzed using the Fischer's exact test. RESULTS: In the prefeedback years (1997 and 1998), high contrast use occurred in 7.7% (n = 602) of the total cases (n = 7799). In the postfeedback years (1999, 2000, and 2001), this declined to 3.6% (n = 445) of the total cases (n = 12,523), representing a 53% reduction (P < .001). The effect was most pronounced in PCI, with the percentage of high contrast cases decreasing from 24% (n = 563) of the total PCI cases (n = 2348) to 10.6% (n = 359) of the total PCI cases (n = 3386), representing a 56% reduction (P < .001). CONCLUSIONS: Our study demonstrates the effectiveness of behavioral modification and the contribution it can make in reducing the volume of contrast dye used in the cardiac catheterization laboratory, particularly in PCI. Adoption of such strategies may enhance patient safety and reduce contribution it can make in reducing the volume of contrast dye used in the cardiac catheterization laboratory, particularly in PCI. Adoption of such strategies may enhance patient safety and reduce costs. PMID- 15354593 TI - Interview with Michael S. Jacobs, JD. PMID- 15354595 TI - Pharmacological treatments for substance use disorders. PMID- 15354596 TI - Homophobia: a challenge for psychosocial nursing. AB - 1. In 1975, the American Psychological Association stated that homosexuality was not a disorder and that homophobia was a form of prejudice based on stereotypes. 2. Because homophobia, like racism or sexism, is learned, it can be unlearned. 3. People who feel homosexuality is a choice hold more negative attitudes toward individuals who are gay, lesbian, bisexual, or transgendered. 4. Homophobia not only damages individuals who are gay, lesbian, bisexual, or transgendered, but also limits heterosexuals by locking them into rigid gender-based roles. PMID- 15354597 TI - Dying patients' thoughts of ending their lives: a pilot study of rural New Mexico. AB - Forty-nine hospice patients in rural New Mexico were directly interviewed concerning their thoughts about ending their lives. Thirty-one patients (63%) did not have thoughts of ending their lives, whereas 18 patients (37%) reported having suicidal thoughts. There were no differences between patients with and without thoughts of suicide related to gender; ethnicity; age; education; disease; religion; importance of religion; location of hospice agency; remaining financial, family, or spiritual issues; satisfaction with hospice care; sum of hospice personnel seen; or sum of medical equipment used. There were significant differences between patients with and without thoughts of suicide related to the number of household members (p = .02); the symptoms of trouble sleeping (p = .04) and nervousness (p =.03); and Medicare insurance coverage for hospice care (p =.01). No other symptom, including pain and hopelessness, was significant. Seven (39%) of the 18 patients who thought of ending their lives told someone about these thoughts. There were no variable differences between patients who did and did not tell someone about these thoughts. PMID- 15354598 TI - A different kind of caregiving support group. AB - 1. Support groups are an important and frequent source of help for caregivers of older adults who experience a high degree of physical and emotional stress. However, not every participant feels comfortable speaking in a group setting. 2. Writing is a form of emotional expression that has been shown to produce both physiological and psychological benefits. Use of narrative writing as an adjunct to traditional group therapy methods has not been studied. 3. In this pilot project, use of writing within the support group format provided caregivers with an additional strategy for coping with the stressors of caregiving. Participants' physical and mental health improved significantly after participating in the group. 4. Therapeutic writing can be used independently or in a group to enable emotional expression. Nurses who work with caregivers can help develop interventions that use narrative writing as a method to improve coping. PMID- 15354599 TI - Immunizations: the greatest good. Pediatricians must advocate for children at the national, state, and local levels. PMID- 15354600 TI - Immunization issues of the 21st century, part I. PMID- 15354601 TI - A 17-year-old boy previously diagnosed with chronic Lyme disease. Patient complained of low-grade fevers, headaches, pharyngitis, and suspected his mother was trying to poison him. PMID- 15354602 TI - One for all: newer combination vaccines in practice. AB - As new combination vaccines are approved, practices must consider whether to switch. The decision is likely to be complex, involving everything from re education of staff to recalculating the bottom line. At some point, however, change will be inevitable, driven principally by the need to add new vaccines to the schedule. In fall 2004, practitioners will be adding two doses of inactivated influenza vaccine to the infant schedule. More communities will be adding two doses of hepatitis A vaccine, which may become routine for all children at some point. We can hope as well for a meningococcal conjugate vaccine series, which, like Hib and PCV-7, would be administered by an infant's first birthday. In addition, there's the pentavalent bovine reassortant rotavirus vaccine (Rotateq) which, although given orally, will further crowd the schedule. The sooner we become comfortable with combination vaccines, the better. PMID- 15354603 TI - Routine infant hepatitis B vaccination recommendations. AB - In the past 2 decades, there have been many changes in vaccine recommendations to prevent hepatitis B. It is important for healthcare providers to understand the importance of universal Hep-B administration to infants, as well as the particulars of delivering the vaccine in practice. Questions will inevitably arise, and practitioners should be equipped to answer those questions with factual information, laying out the very large and tangible potential benefits as well as the theoretical risks. PMID- 15354604 TI - Pertussis is eradication achievable? AB - We are now on the threshold of a major step in reducing the incidence of pertussis and its associated mortality and morbidity in young children. Assuring that vaccines are received in a timely fashion by all children and correcting the "missed opportunities" for providing this highly effective intervention are most critical. A plan for booster immunizations must be developed for older children and adults to offer further protection of young infants through the cocoon effect that this would create for them. PMID- 15354605 TI - Vaccines for rotavirus gastroenteritis universally needed for infants. AB - Rotavirus causes severe and often lifethreatening illness. Universal application of a safe and protective vaccine is justified in both developed and developing nations. Two vaccine candidates, one monovalent (Rotarix) and one multivalent (Rotateq), appear to meet these requirements and are likely to be licensed in the United States in the next 2 or 3 years. Both vaccines exhibited similar safety characteristics. There is little doubt that Rotateq and Rotarix will be shown to be effective for routine protection of infants. Unfortunately, despite numerous clinical trials, the most common serotype (PlaGa) commonly has been encountered as a natural challenge. Therefore, it is not known whether either vaccine possesses advantages in different epidemiological situations. Continuing the analogy with influenza virus, it may be that optimum protection against different serotypes requires a vaccine that is precisely homologous in antigen composition. If so, Rotateq would provide protection against the most common serotype PlaG1 because in includes both Pla and G1 rotavirus reassortants. Further, it would be expected to provide superior protection against G2, G3, and G4 wild-type virus because it contains reassortants of those specificities. In the case of a natural challenge with a serotype that was not G1, G2, G3, or G4, a Rotateq preparation containing a WC3 reassortant expressing the new G serotype could be formulated readily. The monotypic Rotarix may provide ideal protection against the PlaG1 rotavirus because it is composed solely of the PlaG1 strain. It may also provide cross-protection against other rotavirus serotypes adequate to protect against severe and life-threatening disease. In such a case, its monotypic composition may also provide significant economic savings in manufacturing. The resolution of these questions may have to await extensive post-licensure experience with each vaccine. In the future, possible application of rotavirus vaccine for other situations also should be explored, including use in older children to limit nosocomial infection, use in geriatric populations, use in the immunocompromised host, and possibly use in parents and other adults in contact with infants with rotavirus. Both Rotarix and Rotateq likely are to be launched at prices beyond those affordable in the poorest and neediest less-developed countries. It is essential that there be vigorous pursuit of new technologies to manufacture these products at drastically reduced cost if their true lifesaving potential is to be achieved. PMID- 15354606 TI - New vaccines for old diseases: trivalent cold-adapted influenza vaccine. AB - Schoolchildren regularly have the highest influenza attack rates. Schoolchildren also are accessible for vaccination; school-based vaccine clinics can facilitate delivery of vaccine to a high proportion of children in a short period of time. CAIV-T has advantages for immunization of this group due to the effectiveness of a single dose, ease of administration, and ready acceptance by children. Modeling of control for an influenza epidemic by Longini et al. has shown vaccination of 70% of school children would dampen epidemics and significantly reduce the exposure of vulnerable people in the community. The goal of the central Texas trial is to demonstrate this effect in the field. Universal childhood immunization could be an important supplement to current control efforts. Vaccine coverage with TIV has leveled off since 1997; no progress toward the Healthy People 2010 goals of 90% coverage has been made after 4 years in the new decade. The number of excess deaths is expected to double within the next 25 years if control methods are not intensified. CAIV-T may become an important adjunct to control measures and can facilitate influenza pandemic preparedness. PMID- 15354607 TI - Should the perioperative director report to the CNO or to the COO? PMID- 15354608 TI - OR managers facing challenges of gaps in the surgical schedule. PMID- 15354609 TI - Less costly drugs work for nausea, vomiting. PMID- 15354610 TI - Explaining salary sample salary variances variances. PMID- 15354611 TI - Lessons learned from new spine program. PMID- 15354612 TI - Tips on coding and billing for spinal cases. PMID- 15354613 TI - Frequent questions on tissue handling. PMID- 15354614 TI - CDC gives advice on tissue safety. PMID- 15354615 TI - How will surveyors assess competence? PMID- 15354616 TI - Improving odds for implant payments. PMID- 15354617 TI - Patient deaths stark reminder on policies. PMID- 15354618 TI - Cyclosporine A and adverse effects on organs: histochemical studies. AB - The discovery that cyclosporine A (CsA) was a powerful immunosuppressant had a significant impact on transplant medicine. Its molecular mechanism of action has been well defined in T cells and involved inhibition of critical signalling pathways that regulated T-cell activation. In fact, CsA inhibited calcineurin phosphatase activity and thereby activation of the transcription factor nuclear factor of activated T cells. Over 10 years, its use is limited by side effects, determining nephro- and hepatotoxicity, gingival hypertrophy, tremor and increased blood pressure. These negative effects have been identified through morphological alterations and/or clinical parameters, i.e. variation in glomerular filtration rate for nephrotoxicity. Nevertheless, CsA remains a therapeutic valuable agent and it is normally utilized into clinical practice even if different dose adjustments or discontinuations in a significant percentage of patients must be used. This review focuses on the following topics: mechanisms of action and drug metabolism, interactions with other drugs, clinical and morphological evaluation of toxic effects on target organs. In particular, the morphological evaluation of negative effects has been considered reporting light and ultrastructural studies on target organs both in normal and immunosuppressive conditions. Moreover, the histochemical and immunohistochemical variations in cellular metabolism and antigenic properties of cells present in the parenchyma of these organs are discussed. PMID- 15354619 TI - Rethinking literacy and women's health: a Bangladesh case study. AB - Health and literacy are two major areas of women's development in the Third World. Although health and literacy have been recognized as essential elements for improving the quality of women's lives, questions emerge from Eurocentric and colonial assumptions about development, including the following: Does literacy have an impact on women's health? If it does, what are the mechanisms whereby literacy could have an impact on women's health? Using Bangladesh as a case study, I question the Eurocentric and colonial nature of dominant discourses in the answers to questions about literacy and women's health. I argue that literacy and women's health need to be reconsidered from the local women's standpoint since dominant discourses fail to take into consideration local women's worldviews, indigenous knowledge, and oral traditions. Finally, I make some recommendations for future research and programming in literacy and women's health in Bangladesh and in health care in Third World contexts. PMID- 15354620 TI - Work life and psychological health: the experiences of Thai women in deprived communities. AB - I discuss women's work life and the psychological health of women in low-income communities in Northeast Thailand. Previous research has shown that low-income women are part of a disadvantaged group who struggle against several problems in their everyday life, and who work hard to survive. These women worked as either manual laborers in agriculture or factories or as self-employed vendors, and were busy Ha Yoo Ha Kin (working and earning a living). The women's way of life was complex and involved being responsible for their children, husband, extended family, work, and themselves. Understanding women's beliefs and practice relating to work life and health is essential in designing effective intervention programmes to promote the health and well-being of low-income women in Thailand. PMID- 15354621 TI - Guilt and emptiness: women's experiences of miscarriage. AB - Women who lose an early pregnancy are shocked when they are first given the information that they have miscarried. Later they feel guilt and emptiness. Heideggerian interpretive phenomenology has been used with 13 women from southwest Sweden to uncover their lived experience of miscarriage. Women plan their future with a child during early pregnancy. When miscarriage occurs it is not a gore, an embryo, or a fetus they lose, it is their child. They feel that they are the cause of the miscarriage through something they have done, eaten, or thought. They feel abandonment and they grieve for their profound loss; they are actually in bereavement. PMID- 15354622 TI - In-home intervention for depressive symptoms with low-income mothers of infants and toddlers in the United States. AB - Depressive symptoms in low-income mothers negatively affect infant-toddler development. This pilot study tested a short-term, home-based depressive symptom intervention with 16 African American and White, Non-Hispanic mothers in Early Head Start (EHS) programs who were randomly assigned to intervention and usual care/waiting list conditions. Mothers met in their homes with master's-prepared psychiatric mental health nurses who worked with them to improve their management of depressive symptoms and life issues, use of social support, and parenting. The intervention group showed a significantly greater decrease from baseline in depressive symptom severity at 8 and 16 weeks in contrast to the mothers receiving usual care. Observations of maternal interactions showed improvement in the intervention mothers. The results support testing on a larger scale. PMID- 15354623 TI - Strategies of social and sexual control of Malaysian women in psychiatric institutional care. AB - An ethnographic study of female psychiatric patients was undertaken in East Malaysia. Findings indicate that these service users were subject to a number of sexist and oppressive practices that militate against their freedom of movement on the wards as well as their social interaction in the wider context of the hospital. Stereotypic notions of female sexuality and morality act as forms of moral containment and are interpreted in the hospital context as requiring heavy custodial care by the hospital authorities. PMID- 15354624 TI - Demands out of context: Tanzanian women combining exclusive breastfeeding with employment. PMID- 15354625 TI - Ethics and the physician-industry relationship. PMID- 15354626 TI - The evidence thing. AB - The methodology of evidence-based medicine (EBM) has become dominant in the clinical field to the detriment of historically validated paradigms. The philosophical background of EBM is considered and the strengths and flaws of its main tools (randomized trials and meta-analysis) are reviewed. The structural format of EBM speaks the language of regulators and health planners, satisfies the needs of the academic-industrial complex, and is favored by editors of medical journals. Specific problems of trials of medical versus surgical therapy are noted. The deductive-numerical approach of EBM is a methodology geared to ascertain the most effective management of a condition when the differences observed between the two entities being considered is small. EBM methodology is unlikely to lead to discovery of new diseases or treatments; the latter derive from alert observation and inductive inference. While its contribution to the rigorous analysis of effectiveness of some therapies is acknowledged, EBM is not the nostrum of clinical research. Observational and inductive clinical intelligence should be stimulated and published because a therapy needs to be invented before it is proven effective. Biomathematicians need to improve nonrandomized methodology as they did for randomized studies. PMID- 15354627 TI - Computed tomography and ultrasound in follow-up of patients after endovascular repair of abdominal aortic aneurysm. AB - The purpose of this study was to compare our experience with duplex ultrasonography (US) and computed tomography (CT) for the routine follow-up of patients after endovascular repair (EVAR) of abdominal aortic aneurysm (AAA). We reviewed the electronic charts and radiologic exams of the first 125 patients (113 males, 12 females, median age of 76 years, range 48-98 years) with AAA treated by EVAR from June 1996 to November 2001. Our follow-up protocol included serial CT and US at regular intervals after the procedure (before discharge, at 1 month, and then every 6 months). Adequacy of each exam, ability to detect endoleaks, measurements of AAA diameter, and ability to determine graft patency were compared. For endoleak detection, comparison between CT and US was done using CT as the gold standard. A total of 608 exams, 337 CTs and 271 US, were performed 1 day to 5 years after endovascular aneurysm repair; 98% of CT and 74% of US were technically adequate. Contrary to CT, the proportion of adequate US exam was significantly less in patients with higher body mass index (BMI > or = 30 = 54% vs. BMI < 30 = 81%, p < 0.001) and for pre-discharge US compared to the post-discharge US (54% vs. 88%, p = 0.0005). Concurrent scan pairs were obtained in 252 instances in 107 patients (1-8 pairs per patient). Excellent correlation between AAA diameter measured on CT and US was noted (correlation coefficient of 0.9, p < 0.0001). However, agreement was poor. CT anteroposterior (AP) and transverse measurements were on average 2.9 mm (95% limits of agreement = -7 to 13 mm) and 1.8 mm (95% limits of agreement = -9 to 12 mm) greater than US. For AAA diameter change, there was no case of increase AP diameter on CT. However, in 23% (29/128 pairs of sets) of US, an increase in AAA size that could have influenced patient management (> or = 4 mm) was reported despite no change demonstrated on CT. For endoleak detection, sensitivity and specificity of US compared to that of CT was 25% and 89%. Similar sensitivity and specificity were noted when we excluded the first set (25% and 95%), sets done prior to 2000 (30% and 89%), inadequate CT or US scans (31% and 98%), or duplicate sets of results for each patient (28% and 81%). Of the 27 endoleaks missed on US in 17 patients, 2 were type I endoleaks. None of the four endoleaks seen only on US were type I endoleak. US usefulness prior to discharge was reduced by the high rate of inadequate exam, especially in obese patients. Despite the excellent correlation in AAA diameter between US and CT, there was significant disagreement in AAA diameter measurement and diameter change. Sensitivity of nonstandardized US for endoleak was low compared to CT. CT remains our primary imaging study after EVAR, but standardization of post-EVAR US technique may improve its accuracy. PMID- 15354628 TI - Endovascular repair of paraanastomotic aneurysms after previous open aortic prosthetic reconstruction. AB - The aim of this study was to evaluate the effectiveness of endovascular repair of anastomotic and true aortic and iliac aneurysms occurring after prior polyester graft repair for abdominal aortic aneurysms (AAA) or aortoiliac obstructive disease. Between July 1999 and January 2003, 14 patients underwent endovascular treatment of aortic pseudoaneurysms (n = 6) or iliac aneurysms (2 patients with pseudoaneurysms and 6 patients with true aneurysms) occurring 4 to 18.4 years (mean, 8.8 years) after open aortic surgery. No patient had symptoms or positive parameters for infection of the original polyester graft. Eleven patients, including one patient with both a proximal anastomotic and a true iliac aneurysm, were treated with AneuRx (n = 8), Talent (n = 2), or Quantum LP (n = 1) bifurcated stent grafts. Three patients with an infrarenal anastomotic pseudoaneurysm were treated with a tube stent graft (Talent [n = 2] and AneuRx [n = 1]). Endovascular stent grafts were successfully inserted in all patients. Procedure-related complications or death was not seen. During a median follow-up of 12 months (range, 3-40) all anastomotic and/or true aneurysms treated with bifurcated stent grafts maintained excluded. However, two out of three patients, treated with a tube graft for proximal aneurysm exclusion, were converted. In both patients the tube stent graft did not migrate from the level of the renal arteries but fixation failed between the stent graft and the previous polyester graft, creating endotension in the thrombus of the aneurysm sac. In one of these patients the old anastomotic aneurysm ruptured 16 months after stent graft placement and the patient died 1 day after conversion because of mesenterial ischemia. At 1 year follow-up the second patient was converted successfully after enlargement of his anastomotic aneurysm due to similar disconnection between the stent graft and the polyester graft. From this experience with endovascular stent grafts, we conclude that these can be used successfully to exclude anastomotic or true aneurysms after open aortic surgery. Exclusion of aneurysms at the proximal anastomosis with tube stent grafts is apparently not durable because of the insecure distal fixation in polyester grafts. Endovascular repair with bifurcated stent grafts, however, seems to be effective at midterm follow-up. PMID- 15354629 TI - Low quality of life prior to screening for abdominal aortic aneurysm: a possible risk factor for negative mental effects. AB - The objective of this study was to evaluate the effect on quality of life (QOL) of screening for abdominal aortic aneurysm (AAA) in a population-based AAA screening program. Twenty-four patients with screening-detected AAA and 45 controls with normal aortic diameter were studied in a prospective, controlled, population-based study. Prior to and 12 months after the ultrasonography examination, all participants completed Short-Form 36 and at 12 months, 10 AAA specific questions were added. Comparisons were made between the two groups (AAA patients and controls), within each group, and between the groups and norms for the general Swedish population in the same age interval. Our results showed that screening for AAA results in impairment of QOL among those who have the disease and who suffered a low QOL prior to screening. Among those who had an age adjusted normal QOL prior to screening and who were found to have the disease, and among those who were found to have normal aortas, no negative effect on QOL was observed. Thus, low QOL before screening is a possible risk factor for negative mental effects of diagnosing an AAA by screening. PMID- 15354630 TI - A comparison of magnetic resonance angiography, contrast arteriography, and duplex arteriography for patients undergoing lower extremity revascularization. AB - The objective of this study was to compare magnetic resonance angiography (MRA), contrast arteriography (CA), and duplex arteriography (DA) for defining anatomic features relevant to performing lower extremity revascularizations. From March 1, 2001 to August 1, 2001, 33 consecutive inpatients with chronic lower extremity ischemia underwent CA, MRA, and DA before undergoing lower extremity revascularization procedures. The reports of these tests were compared prospectively and the differences in the aortoiliac segment, femoral-popliteal, and infrapopliteal segments were noted. The vessels were classified as mild disease (<50%), moderate disease (50-70%), severe disease (71-99%), and occluded. These studies and treatment plans based on these data were compared. During this time period, 11 patients were not able to undergo MRA and therefore were excluded from the study. Thirty-three patients were included in this study. These patients underwent 35 procedures, as 2 patients underwent bilateral procedures. The mean age of the 33 patients was 76+/-10 years (SD). Indications for the procedures included gangrene (20), ischemic ulcer (8), rest pain (4), and severe claudication (1). Patients' medical history included diabetes mellitus (25), hypertension (20), and end-stage renal disease (5). No differences were noted between intraoperative findings and CA in this series. Two of the three differences between DA and CA were felt to be clinically significant whereas 9 of the 12 differences between MRA and CA were felt to be clinically significant. On the basis of these data in this series, MRA does not yet seem to be able to obtain adequate data on infrapopliteal segments, at least not for this highly selected population. When severe tibial calcification or very low flow states are identified, CA may be necessary for patients undergoing DA. PMID- 15354631 TI - Nicotine induces mononuclear leukocyte adhesion and expression of adhesion molecules, VCAM and ICAM, in endothelial cells in vitro. AB - The pathology of atherosclerotic cardiovascular disease (ASCVD) has been characterized as an inflammatory response to vessel injury. The initial steps of this response involve mononuclear leukocyte (MNL) attachment and infiltration into the vessel wall. Leukocyte adhesion is potentiated by expression of cellular adhesion molecules. Vascular cell adhesion molecule-1 (VCAM) and intracellular adhesion molecule-1 (ICAM) are markers of cellular activation and have the ability to attach leukocytes to the endothelium, which is an initial event in the inflammatory response in the vessel wall. Human umbilical vein endothelial cells (HUVEC) were plated in endothelial growth medium (EGM) on plastic coverslips and grown until cells were 75% confluent. Free base nicotine (FBN) was diluted in EGM to a concentration of 10(-8) M and added to experimental cells. At 3 hr, coverslips were removed and fixed. Immunohistochemical staining (IHCS) was performed using a monoclonal antibody to human ICAM and VCAM. Digital image analysis (DIA) was performed to quantify the expression of ICAM and VCAM. An intensity stain index (ISI) measuring area and intensity of stain/total cellular area was determined. Additional HUVEC grown in a similar manner were either exposed to 10(-8) M FBN in EGM or EGM control for 4 hr, then were exposed to MNL suspension for 10 min. Coverslips were removed, rinsed, and fixed. Hematoxylin and eosin staining was performed and cells examined under light microscopy. Leukocyte number per high power field (HPF) was counted and compared to controls. Data were analyzed using analysis of variants (ANOVA) and Student's t-test. Differences were considered significant if p < 0.05. ICAM and VCAM expression was absent in control cells. Nicotine exposure at 3 hr induced expression of VCAM (ISI = 30.85+/-0.77) and to a lesser extent ICAM (ISI = 16.6+/-1.39) (p < 0.001). MNL adhesion was markedly increased in cells exposed to nicotine (79.4+/ 16.9/HPF) when compared to control cells (1.8+/-0.91/HPF) exposed to MNL (p < 0.01). These data show nicotine's ability to activate HUVEC as evidenced by induction of ICAM and VCAM expression in vitro. The biological effects of these adhesion molecules are demonstrated by a marked increase in MNL adhesion to HUVEC as demonstrated by leukocyte adhesion assay (LAA). MNL adhesion and subsequent migration into the intima, if occurring in vivo, may be a vital step in the pathogenesis of ASCVD associated with nicotine exposure. PMID- 15354632 TI - An in vitro model to compare the antimicrobial activity of silver-coated versus rifampicin-soaked vascular grafts. AB - In situ replacement of infected vascular grafts is an accepted alternative to total graft excision and extraanatomic replacement. Its success relies upon the ability of the newly inserted graft to resist recurrent infection. This study compares the efficacy of two methods used to reduce the risk of graft reinfection: rifampicin soaking versus silver bonding of grafts. The grafts' resistance to infection was tested in vitro in two protocols, each using a panel of seven common bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The length of time the grafts remained free of organisms was compared between the groups. Both the silver graft and the rifampicin-soaked graft were significantly better than control graft at preventing bacterial growth on the graft surface. The rifampicin inhibited the growth of the gram-positive organisms, including MRSA, significantly better than the silver graft on days 2 and 3 (p < 0.001). Conversely, the silver graft was significantly more effective against the gram-negative organisms until day 4 (p < 0.0001). Both types of graft inhibit the in vitro growth of bacteria more effectively than controls, with rifampicin being most effective against gram-positive organisms and silver being best against the gram-negative organisms. PMID- 15354633 TI - Thrombolysis as initial treatment of peripheral native artery and bypass graft occlusions in a general community hospital. AB - Large series with long-term follow-up of thrombolytic therapy in the treatment of lower limb arterial occlusion from a single, general, non-university hospital are absent. We studied retrospectively the results of 129 consecutive patients who underwent thrombolysis with intraarterial urokinase as initial treatment for lower limb native artery or bypass graft occlusions. The mean age of patients was 71 years; 55% of the patients were male, and preexisting peripheral arterial disease was present in 47%. Presenting symptoms were disabling claudication (31%) and limb-threatening ischemia (69%). Forty-two percent of the patients presented with acute symptoms (<1 week duration). The mean follow-up of patients still alive at the time of analysis was 36 months (range 1-120 months). Thrombolytic treatment was successful in 93 patients (72%). In 53% of the patients acute surgical intervention could be avoided: 28 patients (22%) did not need any additional procedure and 40 (31%) underwent a concomitant angioplasty. When thrombolysis failed, 6 patients (5%) underwent successful surgical revascularization and 11 patients (8%) eventually underwent major amputation during their hospital stay. Amputation-free survival at 6 months and at last follow-up was 88% and 83%, respectively. The mortality rates were 4% at 30 days, 5% at 6 months, and 30% at last follow-up. Thrombolysis was significantly less successful when patients had diabetes (62% vs. 81%, p = 0.019) or preexisting peripheral arterial disease (61% vs. 80%, p= 0.018). Successful radiological treatment (thrombolysis+/-angioplasty) could less often be performed in patients with preexisting peripheral arterial disease (41% vs. 59%, p = 0.011) and in patients with occluded bypass grafts (33% vs. 62%, p= 0.002). Duration of symptoms and Fontaine stage at presentation did not predict thrombolysis outcomes. Thrombolytic-related complications occurred in 17 patients (13%), with significant bleeding from the puncture site in 3 patients (2%). Thrombolysis can safely and effectively be performed in a general community hospital with results comparable to those reported from specialized university centers and large randomized trials. PMID- 15354634 TI - Popliteal-to-distal bypass for limb salvage. AB - A retrospective study was carried out to examine the patency and limb salvage rates of popliteal-to-distal bypass and compare the results of diabetic to those of nondiabetic patients and elective versus emergency procedures. From January 1990 to December 2001, 71 popliteal-to-distal bypasses were performed. Indications for surgery were rest pain, tissue loss, and acute ischemia, including extensive post-traumatic tibial lesions. Survival, graft patency, and limb salvage rates were determined according to the life-table method. The log rank test was used to compare diabetic versus nondiabetic patients, elective versus emergency procedures, and saphenous vein bypass versus PTFE bypass. Postoperative primary patency, secondary patency, and limb salvage rates at 30 days were 88.7%, 91.4%, and 87%, respectively. Postoperative mortality was 2.8%, with one case of acute myocardial ischemia and one multiorgan failure. Life-table analysis showed primary and secondary patency rates of 57% and 61%, respectively, a limb salvage rate of 64%, and survival of 77% at 5 years Log-rank testing showed no statistical difference between diabetic and nondiabetic patients, whereas a statistical difference was observed in elective versus emergency procedures (p < 0.005) and great saphenous vein versus PTFE graf (p < 0.05). These results show that popliteal-to-distal bypass is a safe and effective procedure with good long-term patency and limb salvage rates in selected cases. PMID- 15354635 TI - Expanding options for bedside placement of inferior vena cava filters with intravascular ultrasound when transabdominal duplex ultrasound imaging is inadequate. AB - Although bedside placement of inferior vena cava (IVC) filters by means of transabdominal duplex ultrasound is possible in most patients, those with inadequate visualization have traditionally required fluoroscopy. The purpose of this study was to assess the safety and efficacy of bedside IVC filter placement with intravascular ultrasound (IVUS) when transabdominal duplex ultrasound imaging is inadequate. Between January 1, 1999 and December 31, 2002, 256 IVC filter placements were performed (207 with transabdominal duplex ultrasound [81%], 21 with fluoroscopy [8%], and 28 with IVUS [11%]). IVC filter placement with IVUS was performed only if visualization with transabdominal duplex ultrasound was determined to be inadequate. Demographics, technical data, and outcome for patients undergoing IVC filter placement with IVUS were reviewed and form the basis of this report. Bedside IVC filter placement with IVUS was technically successful in 26 of 28 patients (93%). Post-procedure abdominal radiographs confirmed proper placement, based on bony landmarks in 24 of 26 patients (92%). Post-procedure complications included insertion site thrombosis in two patients and possible recurrent pulmonary embolism in one patient 2 months following filter placement. One patient died from causes unrelated to IVC filter placement. From these results we conclude that IVC filter placement with IVUS is technically feasible and safe. This may allow for expanded bedside IVC filter placement capabilities in patients with inadequate IVC visualization on transabdominal duplex ultrasound. PMID- 15354636 TI - Descending thoracic and thoracoabdominal aortic aneurysm repair: 15-year results using a uniform approach. AB - This review presents the results of surgical repair of descending thoracic (DT) and thoracoabdominal aortic (TAA) aneurysms, using spinal drainage (SD) distal aortic perfusion (DAP), and other adjuncts intended to reduce complications. Records of patients undergoing repair of DT and TAA between 1986 and 2002 were reviewed. Elective operations were performed using single lung ventilation, invasive monitoring, SD, modest anticoagulation, permissive hypothermia (> or = 33 degrees F), liberal use of transaortic endarterectomy, and complete repair. Intercostal arteries were reimplanted when possible and DAP was used in DT and TAA types I, II, and III repair. Exceptions to this approach were noted. Some of these adjuncts were used in emergency cases. Actuarial survival was calculated. Fifty consecutive patients with DT (3) or TAA (47), type I (4), type II (16), type III (18), or type IV (9), aneurysms received elective (36) or emergency (14) repair between 1986 and 2002. Mortality was 2/36 (5.5%) in the elective group. In the emergency group, there were 2 intraoperative deaths and mortality was 4/14 (28.5%, p < 0.07). Overall survivor morbidity was 6/34 (17.6%) in elective and 7/10 (70%, p < 0.02) in emergency cases. Paraplegia occurred in one patient in the elective group (2.7%) with dissecting type II TAA aneurysm in whom the intercostal patch was sacrificed. Two of 12 initial survivors developed paraplegia in the emergency group (16.7%); one had SD but neither had DAP or intercostal reimplantation. Serious complications were associated with avoidable deviations from the approach. Five and 10-year survival for the entire series was 64.8% and 46.4%, respectively. These results parallel those in contemporary reports from centers where repair of descending and thoracoabdominal aortic aneurysm is frequently performed. Good long-term results can be achieved using spinal drainage and distal aortic perfusion, combined with other adjuncts as a means of reducing complications. When possible, the same approach should be used in emergency cases. PMID- 15354637 TI - Prevention of postischemic spinal cord injury by means of regional infusion of adenosine and L-carnitine dissolved in normothermic saline. AB - Spinal cord ischemia still remains an unsolved problem in modern aortic surgery. In this study, we investigated the effectiveness of combined agents such as adenosine and L-carnitine infused to the isolated segment of abdominal aorta in a rabbit model. Twenty-eight rabbits divided into four groups underwent 40 min of isolated infrarenal aortic occlusion. Group I animals received no medication. Group II received an infusion of 100 mg/kg L-carnitine in normothermic saline over the first 10 min of ischemia. Group III received 50 mg adenosine, and group IV received a combination of the two agents in the same fashion. Spinal cord function was evaluated at 24 and 72 hr after operation on the basis of Tarlov scale and similar results were obtained. After a second evaluation, spinal cords were harvested for histological examination. Group I animals were all paraplegics. Spinal cord function was partially intact in two of the group II animals with Tarlov scores of 5 in two and 4 in two whereas one of the rabbits could not hop with a score of 3, and the remaining two could not sit with scores of 1 and 0. The spinal cord function of group III animals was intact with Tarlov scores of 5 in three, 4 in two, and 3 and 1 in remaining ones. In the group IV animals, it was fully intact with Tarlov scores of 5. Histological examination in group I revealed marked enlargement of the vacuoles of glial cells in the white matter of spinal cord. Glial cells were deteriorated in some locations in group II whereas they were mostly protected in the third group. In group IV, histological examination revealed no evidence of spinal cord injury. In conclusion, combined infusion of adenosine and L-carnitine provided better protection against postischemic spinal cord injury than individual infusion of these agents. PMID- 15354638 TI - Severe coagulopathy following intraoperative use of topical thrombin. AB - This report describes a case of severe coagulopathy and bleeding related to the intraoperative use of topically applied thrombin. Commercial thrombin preparations contaminated with bovine factor V have been shown to stimulate the production of antibodies directed against factor V. These antibodies can cause coagulopathy. Our patient developed antibodies against factor V after intraoperative exposure to topical thrombin. The resulting antibody-mediated depletion of factor V caused a severe and refractory coagulopathy. Vascular surgeons should be aware that the use of topical bovine thrombin can cause severe coagulopathy. PMID- 15354639 TI - Endoscopic diagnosis of a clinically silent aortoesophageal fistula: case report and review of the literature. AB - We report a rare case of a secondary aortoesophageal fistula discovered incidentally during elective upper endoscopy. The patient had previously undergone repair of a descending thoracic aortic aneurysm with a Dacron interposition graft. Esophagoscopy 2 months after the aneurysm repair demonstrated a large mid-esophageal erosion with visualization of the aortic graft at the base. The aortoesophageal fistula had been clinically silent to this point. During preparation for surgery the patient developed large-volume esophageal hemorrhage and died following attempted endovascular repair of the fistula. A review of the literature on the diagnosis and surgical management of aortoesophageal fistula is presented. PMID- 15354640 TI - Endovascular repair of inflammatory abdominal aortic aneurysms: a valuable alternative?--Case report and review of literature. AB - While endovascular repair (ER) has become a routine procedure in the treatment of arteriosclerotic abdominal aortic aneurysms with a suitable configuration, only rare reports of interventional treatment of inflammatory aortic abdominal aneurysms (IAAA) exist. We present a case study of a male patient with IAAA, who presented with inflammatory thickening involving the entire circumference of the aortic vessel wall. The MRI performed 8 months after successful ER demonstrated complete regression of vessel wall induration. A patient with the appropriate anatomical configuration of IAAA should benefit from the lower morbidity and mortality of endovascular aneurysm repair (EVAR). In our view, EVAR is preferable to open surgical repair in the specific situation of IAAA. PMID- 15354641 TI - Aortobipopliteal bypass grafting for in situ replacement of infected aortobifemoral prosthesis. AB - Conventional treatment of an infected aortobifemoral graft includes total graft excision and ex situ bypass grafting, but has been associated with significant perioperative morbidity and mortality. Additionally, the presence of infection in the groin makes limb revascularization problematic. There is increasing evidence that in situ replacement of an infected graft can achieve promising results in selected patients. We present a case of an aortobifemoral graft infection, affecting both the groin as well as the entire pelvis. The patient underwent successful in situ graft replacement with a new aortobipopliteal prosthesis via an alternative extraperitoneal route. PMID- 15354642 TI - Salvage of the hand with a free flap in a hemodialysis patient with finger gangrene and ipsilateral arteriovenous fistula: a case report. AB - We describe the case of a female with end-stage renal disease who developed a left ischemic hand and index finger gangrene 2 years after commencement of hemodialysis. Ray amputation was performed, however, the surgical wound was complicated by progressive soft tissue necrosis and metacarpal bones exposure. To avoid amputation at a higher level, we repaired the defect successfully with a free anterolateral thigh perforator flap. A radiocephalic arteriovenous fistula, the dialysis access, was divided to provide the recipient vessels. Reconstruction with this method has the advantage of a staged arteriovenous fistula, which can increase the free-flap success rate, and has the potential to improve perfusion of the distal ischemic hand. PMID- 15354643 TI - Compression of the iliac vein by a synovial cyst. AB - While a synovial cyst of the hip is a rare cause of lower limb swelling, a differential diagnosis of deep vein thrombosis is needed. A case of lower limb swelling due to compression of the external iliac vein by a synovial cyst is presented with a review of the literature. Sagittal views and three-dimensional images of a CT scan were very useful for the diagnosis. This is the first review in English of vessel compression by a synovial cyst of the hip. PMID- 15354644 TI - Successful treatment of infected vascular prosthetic grafts in the groin using conservative therapy with povidone-iodine solution. AB - Four cases of infected vascular prosthetic graft in the groin successfully treated with povidone-iodine solution using a conservative approach are described here. In all patients the same technique was used. After complete debridement, the prosthetic graft in the groin was completely exposed. The wound was cleansed with hydrogen peroxide and then dressed with gauze soaked in 1:10 sterile water diluted povidone-iodine solution. The dressings were changed twice a day. The patients were supplemented by systemic therapy of an appropriate antibiotic. All patients were observed in the intensive care unit. In all patients this treatment method led to control of infection and healing of the wound. Thus, it was not necessary to remove the prosthetic graft and patients were spared a major surgical intervention. At follow-up, the prosthetic grafts remain patent without any signs of recurrence of infection. PMID- 15354645 TI - Covered stent exclusion of blunt traumatic carotid artery pseudoaneurysm: case report and review of the literature. AB - Injury to the carotid artery from blunt trauma, when not lethal, will often go unrecognized. A 37-year-old male was the belted driver in a motor vehicle accident 15 years prior to presenting with intermittent left upper extremity parasthesias and weakness. A large, calcified distal right internal carotid artery (ICA) pseudoaneurysm was diagnosed and successfully excluded with a 7 x 30 mm covered stent. PMID- 15354646 TI - Critical issues in management of congenital vascular malformation. AB - Congenital vascular malformation (CVM) has long been a challenging problem. Recently, a new classification of CVMs was launched, opening up a new era. This improved definition provides critical information on CVM etioanatomopathophysiology. Advanced diagnostic technology has also provided more accurate and safer diagnosis. Finally a new, improved method of managing CVMs based on a new, multidisciplinary approach has emerged. In this study, various noninvasive diagnostic tests were adopted for contemporary diagnosis, and invasive studies were reserved for use as a road map for treatment. A new treatment strategy evolved, following proper identification of absolute and relative indications for treatment, which integrates surgical therapy and embolosclerotherapy. Three hundred and sixty-two patients selected for treatment, among a total of 1007 patients with various indications, were retrospectively reviewed. Various embolosclerotherapy techniques were performed on 333 patients, including 640 sessions of ethanol-based therapy. The immediate success rate was excellent (96.2%) but was accompanied by various complications mostly of a minor nature during 212 sessions (22%). The interim results were also excellent, with no evidence of recurrence. A total of 102 patients underwent 128 surgical procedures, consisting of a variety of different therapies, either independently (29/102) or in conjunction with preoperative embolosclerotherapy (73/102). Most of the interim results were excellent (111/128). The use of a complex therapy regimen through a multidisciplinary approach leads to a more manageable and satisfactory outcome. PMID- 15354647 TI - The dentist's dilemma. PMID- 15354648 TI - Frustration: educating dental professionals to provide care to people with special needs. PMID- 15354649 TI - Gloves in the dental office: their use and effectiveness. PMID- 15354650 TI - Steps to redesign smiles using current technology and materials. PMID- 15354651 TI - Aesthetic restoration of the anterior maxilla. PMID- 15354652 TI - A technique for veneer temporization. PMID- 15354653 TI - Reliable cementation technique for CAD/CAM restorations. PMID- 15354654 TI - Improving accuracy and simplifying treatment with full arch removable prosthetics: a case report. PMID- 15354655 TI - Rotary instrumentation: asking the right questions, part 1. PMID- 15354656 TI - The treatment planning consultation: the doctor/technician partnership. PMID- 15354657 TI - Creating natural gingival profiles using the ovate pontic technique. PMID- 15354659 TI - Indications and techniques for surgical indexing at stage I implant surgery. PMID- 15354658 TI - The functional and aesthetic indications for soft-tissue (gingival) grafting. PMID- 15354660 TI - The eleven essentials of a staff meeting-centered dental practice, part 1. PMID- 15354661 TI - Ten administrative tasks for your computer network. PMID- 15354662 TI - President' message. Address to the Texas Dental Association House of Delegates, May 13, 2004. PMID- 15354663 TI - Trustee's message. Address to the Texas Dental Association House of Delegates, May 13, 2004. PMID- 15354665 TI - Preserving oral health in cancer therapy. PMID- 15354664 TI - Dr. Blanton responds to a letter to the editor. PMID- 15354666 TI - Recurrent human papilloma virus associated lesions in an immunocompromised host: a case report. PMID- 15354667 TI - Xerostomia management in the head and neck radiation patient. PMID- 15354669 TI - So you thought your dental school was tough? PMID- 15354668 TI - Management of oral mucositis pain. AB - Effective pain control for mucositis requires constant attention and willingness on the part of managing clinicians to evaluate and adapt pain relieving strategies throughout the period of risk for oral mucositis. By utilizing the principles of a individualized, tiered approach to pain management, that address the multidimensional components of a patient's pain, maximum comfort can be consistently provided while reducing the risk for side-effects. PMID- 15354670 TI - Empathy. PMID- 15354671 TI - Oral and maxillofacial pathology case of the month. Mucoepidermoid carcinoma. PMID- 15354672 TI - Real-time solutions provide real benefits for TDA member dentists. PMID- 15354673 TI - Current trends for the management and treatment of depression. PMID- 15354674 TI - Managing antidepressant drug therapy in an evolving marketplace. AB - The availability of generic SSRI antidepressants has simplified formulary decisions related to how best to treat uncomplicated depression. However, several new challenges have arisen. Better data are needed to guide formulary guidelines concerning the reuse of nonpreferred antidepressants for patients with recurrent episodes of depression and the risks associated with switching long-term patients to generic SSRIs. Managed care organizations need more information on the appropriateness of expanding the use of generic SSRIs to treat less severe forms of depression. An expansion of the use of antidepressant therapy will likely reduce HEDIS compliance performance, which could adversely affect the marketing of the plan to potential enrollees. Plans must also consider FDA concerns that the newer antidepressants may increase suicidality in a small number of patients. Finally, more research is needed to determine if the generic SSRIs adequately meet the therapeutic needs of more severely ill patients, such as treatment refractory depressed patients and patients with bipolar disorders. PMID- 15354675 TI - Recent developments in antidepressant therapy in special populations. AB - Major depressive disorder (MDD) in the elderly, children, and patients with unstable angina or acute myocardial infarction (MI) is a serious disorder. In elderly patients, where major depression is often overlooked and may have a significant impact on the quality of life, the goal of therapy is full remission. Selective serotonin reuptake inhibitors (SSRIs) have the most favorable combination of efficacy and side-effect profile for the elderly with MDD, regardless of the presence of medical comorbidities. Although the dual agent venlafaxine has been proposed as an alternative agent for older patients who are either nonresponders or partial responders to SSRIs, the frail elderly may be particularly vulnerable to its side effects. Most elderly patients have a relapse of depression when antidepressants are stopped; depression subsides when antidepressants are resumed. Because recent evidence suggests that the dosage of an antidepressant that achieves remission in the elderly does not always protect against recurrence, in addition to long-term maintenance, consideration should be given to increased dosage. Patients with major depression and either unstable angina or acute MI should be identified and considered for antidepressant treatment. Findings from the recent Sertraline Anti-Depressant Heart Attack Randomized Trial suggest that SSRIs may have antiplatelet and endothelium protective properties that may benefit patients with depression and comorbid coronary artery disease and ischemic stroke. Concern regarding the safety of SSRIs in children has prompted new studies. Evidence suggests that the risks of SSRIs, except for fluoxetine, might outweigh benefits in the treatment of depression in children and adolescents. PMID- 15354676 TI - How drug-drug interactions can impact managed care. AB - DDIs have important implications for managed care. They can adversely impact patient outcomes and can also have serious cost implications. The problem posed by unintended and adverse DDIs will continue to grow in importance as the complexity and uniqueness of medication regimens escalate in an aging population using increasingly sophisticated and varied pharmacological agents. In this paper, antidepressants and SSRIs in particular were used to illustrate these important points. PMID- 15354677 TI - Management and treatment of reversible airway disease. PMID- 15354678 TI - Epidemiology, clinical and economic burden, and natural history of chronic obstructive pulmonary disease and asthma. AB - Chronic obstructive pulmonary disease (COPD) and asthma are conditions that exact a tremendous toll on patients, providers, and society. The substantial increase in the prevalence of both conditions in recent decades has generated sizable concern from both domestic and global perspectives. The underlying characteristics of both conditions involve inflammation of the respiratory tract, although the specific nature and reversibility of these processes differ according to each illness. Within the context of disease management, acute exacerbations are important clinical events that contribute to an increase in morbidity and mortality, and may occur in any patient suffering from the disease. Because these conditions are highly important to clinical practice and healthcare systems, this article will highlight key aspects of epidemiology, burden of illness, and clinical presentation of COPD and asthma. A review of the definition, classification, and natural history is also offered, emphasizing the role of acute exacerbations. In general, the natural history of both COPD and asthma is highly variable and not precisely defined because of their heterogeneous clinical courses. Continued inquiry concerning the epidemiology, etiology, classification, and prognosis of each condition and related exacerbations may offer clinicians improved decision-making information to optimize interventions for affected patient populations. PMID- 15354679 TI - An assessment of therapeutic regimens in the treatment of acute exacerbations in chronic obstructive pulmonary disease and asthma. AB - Acute exacerbations in chronic obstructive pulmonary disease (COPD) and asthma are potentially life-threatening clinical events that may result in substantial morbidity and mortality. Treatment of these episodes requires the rapid reversal of airway obstruction by decreasing bronchoconstriction and inflammation. Consensus guidelines and recommendations build on stepwise approaches to care, with the mainstay of therapeutic interventions involving brochodilators and often including systemic corticosteroids, oxygen, and other treatments, depending on severity and setting. Future therapies that target inflammatory processes may offer improved efficacy and potential disease-modifying effects. The purpose of this article is to assess the treatment options for acute exacerbations in COPD and asthma within the scope of current consensus guidelines and recommendations (eg, Global Initiative for Chronic Obstructive Lung Disease, National Asthma Education and Prevention Program, Global Initiative for Asthma). Although bronchodilators and corticosteroids are the primary therapies discussed, antibiotics, oxygen, magnesium sulfate, noninvasive positive pressure ventilation, and helium/oxygen mixtures are also addressed. Preventive approaches for future exacerbations are considered in the overall approach to achieve optimal outcomes. PMID- 15354680 TI - The cost effectiveness of levalbuterol versus racemic albuterol. AB - Albuterol is a selective beta2-agonist that is widely used in the prevention and treatment of reactive airway disease. It is formulated as a racemic mixture containing equal parts of the R- and S-isomers. The therapeutic activity of albuterol is due entirely to the R-isomer, whereas the S-isomer may actually have detrimental effects. Because the slowly metabolized S-isomer tends to accumulate in the body, there has been concern that chronic use of racemic albuterol might lead to loss of effectiveness and clinical deterioration, with potentially serious health and cost consequences. Levalbuterol is a formulation containing only the R-isomer of albuterol, and clinical trials have demonstrated that it offers therapeutic advantages over racemic albuterol. The cost effectiveness of levalbuterol derives mainly from reduced need for acute medical care and hospitalization. PMID- 15354681 TI - Issues and challenges for managed care in the treatment of asthma and chronic obstructive pulmonary disease. AB - Managed care organizations (MCOs) and payers have become increasingly aware of the impact of asthma and chronic obstructive pulmonary disease on healthcare expenditures. The prevalence and incidence of both conditions continue to increase, even as new therapeutic modalities enter the market to offer treatment alternatives for these conditions. Although viewed by most payers as long-term conditions with expenditures that will not diminish, MCOs and payers have become more concerned with controlling costs and ensuring appropriate utilization. PMID- 15354682 TI - A new breed of computer users: rats control a cursor via joystick manipulation. AB - Many exciting findings have been reported in the 15 years since monkeys were first demonstrated to have the capacity to perform computerized tasks. The present data indicate that albino rats can also learn to respond to computer generated stimuli by manipulating a joystick. Although the rat's control of the cursor is not as skillful as has been reported for primate species, it is clearly better than chance and suggests the great potential for comparative investigation afforded by use of the computer test system. PMID- 15354683 TI - AutoTutor: a tutor with dialogue in natural language. AB - AutoTutor is a learning environment that tutors students by holding a conversation in natural language. AutoTutor has been developed for Newtonian qualitative physics and computer literacy. Its design was inspired by explanation based constructivist theories of learning, intelligent tutoring systems that adaptively respond to student knowledge, and empirical research on dialogue patterns in tutorial discourse. AutoTutor presents challenging problems (formulated as questions) from a curriculum script and then engages in mixed initiative dialogue that guides the student in building an answer. It provides the student with positive, neutral, or negative feedback on the student's typed responses, pumps the student for more information, prompts the student to fill in missing words, gives hints, fills in missing information with assertions, identifies and corrects erroneous ideas, answers the student's questions, and summarizes answers. AutoTutor has produced learning gains of approximately .70 sigma for deep levels of comprehension. PMID- 15354684 TI - Coh-metrix: analysis of text on cohesion and language. AB - Advances in computational linguistics and discourse processing have made it possible to automate many language- and text-processing mechanisms. We have developed a computer tool called Coh-Metrix, which analyzes texts on over 200 measures of cohesion, language, and readability. Its modules use lexicons, part of-speech classifiers, syntactic parsers, templates, corpora, latent semantic analysis, and other components that are widely used in computational linguistics. After the user enters an English text, CohMetrix returns measures requested by the user. In addition, a facility allows the user to store the results of these analyses in data files (such as Text, Excel, and SPSS). Standard text readability formulas scale texts on difficulty by relying on word length and sentence length, whereas Coh-Metrix is sensitive to cohesion relations, world knowledge, and language and discourse characteristics. PMID- 15354685 TI - Automatic classification of dysfunctional thoughts: a feasibility test. AB - The identification of dysfunctional thoughts is a central effort in cognitive therapy. This paper describes the first version of a computer module that classifies dysfunctional thoughts automatically. It is part of COGNO, a system we are developing to give automatic feedback on dysfunctional thoughts. The system uses rules that were developed from language markers identified in a sample of 149 dysfunctional thoughts. The system was tested with an independent set of 112 example thoughts. The system detects the majority of dysfunctional thoughts, but works reliably only for some thought categories. Automatic thought classification may be a first step toward developing natural dialogue systems in cognitive therapy. PMID- 15354686 TI - Identifying reading strategies using latent semantic analysis: comparing semantic benchmarks. AB - We explored methods of using latent semantic analysis (LSA) to identify reading strategies in students' self-explanations that are collected as part of a Web based reading trainer. In this study, college students self-explained scientific texts, one sentence at a time. ISA was used to measure the similarity between the self-explanations and semantic benchmarks (groups of words and sentences that together represent reading strategies). Three types of semantic benchmarks were compared: content words, exemplars, and strategies. Discriminant analyses were used to classify global and specific reading strategies using the LSA cosines. All benchmarks contributed to the classification of general reading strategies, but the exemplars did the best in distinguishing subtle semantic differences between reading strategies. Pragmatic and theoretical concerns of using LSA are discussed. PMID- 15354687 TI - iSTART: interactive strategy training for active reading and thinking. AB - Interactive Strategy Training for Active Reading and Thinking (iSTART) is a Web based application that provides young adolescent to college-age students with high-level reading strategy training to improve comprehension of science texts. iSTART is modeled after an effective, human-delivered intervention called self explanation reading training (SERT), which trains readers to use active reading strategies to self-explain difficult texts more effectively. To make the training more widely available, the Web-based trainer has been developed. Transforming the training from a human-delivered application to a computer-based one has resulted in a highly interactive trainer that adapts its methods to the performance of the students. The iSTART trainer introduces the strategies in a simulated classroom setting with interaction between three animated characters-an instructor character and two student characters-and the human trainee. Thereafter, the trainee identifies the strategies in the explanations of a student character who is guided by an instructor character. Finally, the trainee practices self explanation under the guidance of an instructor character. We describe this system and discuss how appropriate feedback is generated. PMID- 15354688 TI - The DIAGNOSER project: combining assessment and learning. AB - DIAGNOSER is an Internet-based tool for classroom instruction. It delivers continuous formative assessment and feedback to high school physics students and their teachers about the correct and incorrect concepts and ideas the students may hold regarding physical situations. That is, it diagnoses misconceptions that underlie wrong answers of students, such as a confusion of velocity with acceleration. We use data about patterns of student responses, particularly consistency of errors from question to question, to improve the system's understanding of student concepts. PMID- 15354689 TI - Human use regulatory affairs advisor (HURAA): learning about research ethics with intelligent learning modules. AB - The Human Use Regulatory Affairs Advisor (HURAA) is a Web-based facility that provides help and training on the ethical use of human subjects in research, based on documents and regulations in United States federal agencies. HURAA has a number of standard features of conventional Web facilities and computer-based training, such as hypertext, multimedia, help modules, glossaries, archives, links to other sites, and page-turning didactic instruction. HURAA also has these intelligent features: (1) an animated conversational agent that serves as a navigational guide for the Web facility, (2) lessons with case-based and explanation-based reasoning, (3) document retrieval through natural language queries, and (4) a context-sensitive Frequently Asked Questions segment, called Point & Query. This article describes the functional learning components of HURAA, specifies its computational architecture, and summarizes empirical tests of the facility on learners. PMID- 15354690 TI - COR V2: teaching observational research with multimedia courseware. AB - Courseware for Observational Research (COR Version 2) is an interactive multimedia program designed to teach the foundation of the scientific method: systematic observation. COR uses digital video with interactive coding to teach basic concepts, such as creating precise operational definitions; using frequency, interval, and duration coding; developing sampling strategies; and analyzing and interpreting data. Through lessons, a case study, and laboratory exercises, it gradually scaffolds students from teacher-directed learning into self-directed learning. The newest addition to COR is a case study in which students work collaboratively, using their own observations to make recommendations about a child's disruptive behavior in an after-school program. Evaluations of the lessons showed that classes using COR received better grades on their field observations than did those using methods that are more traditional. Students' confidence and knowledge increased as they moved through each section of the program. PMID- 15354691 TI - VIZ: the visualization assessment and training web site. AB - The VIZ Website (http://viz.bd.psu.edu/viz/) was developed as a free and open portal for assessment and training of spatial skills. Using Macromedia Authorware, it allows the collection of response times and accuracies from four spatial tasks: mental rotation, paper folding, water level judgments, and spatial working memory. Additional tasks are under development. The site has been used for a variety of teaching and research purposes. For example, incoming engineering students have completed the tasks to assess their spatial skills before beginning their graphics courses. Those with poorer skills can then receive the appropriate remedial instruction. The VIZ site has also been used to teach about spatial cognition and to do research outside the laboratory setting. Finally, we are using the site to conduct basic research on spatial cognition, including an examination of the factors that may reduce gender differences in performance. PMID- 15354692 TI - A web-based family problem-solving intervention for families of children with traumatic brain injury. AB - We developed a Web-based intervention for pediatric traumatic brain injury (TBI) and examined its feasibility for participants with limited computer experience. Six families, including parents, siblings, and children with TBI, were given computers, Web cameras, and high-speed Internet access. Weekly videoconferences with the therapist were conducted after participants completed on-line interactive experiences on problem solving, communication, and TBI-specific behavior management. Families were assigned to videoconference with NetMeeting (iBOT cameras) or ViaVideo. Participants ranked the Web site and videoconferences as moderately to very easy to use. ViaVideo participants rated videoconferencing significantly more favorably relative to face-to-face meetings than did NetMeeting participants. Both the Web site and videoconferencing were rated as very helpful. All families demonstrated improved outcomes on one or more target behaviors, including increased understanding of the injury and improved parent child relationships. All parents and siblings and all but 1 child with TBI said they would recommend the program to others. We conclude that a face-to-face intervention can be successfully adapted to the Web for families with varied computer experience. PMID- 15354693 TI - Tracking attention with the focus-window technique: the information filter must be calibrated. AB - Many researchers have recommended the focus-window tracking (FWT) technique to study the course of information acquisition and visual attention. In FWT, the stimulus is blurred, and the subject scans for information by moving a clear window with a computer mouse. We show that poor calibration of the blur filter will bias search behavior in FWT because diagnostically useful information may leak through a filter even though the subjects are unaware of it. To provide a valid index of information acquisition, FWT requires an information filter that is calibrated to the task. PMID- 15354694 TI - QMPE: estimating Lognormal, Wald, and Weibull RT distributions with a parameter dependent lower bound. AB - We describe and test quantile maximum probability estimator (QMPE), an open source ANSI Fortran 90 program for response time distribution estimation. QMPE enables users to estimate parameters for the ex-Gaussian and Gumbel (1958) distributions, along with three "shifted" distributions (i.e., distributions with a parameter-dependent lower bound): the Lognormal, Wald, and Weibul distributions. Estimation can be performed using either the standard continuous maximum likelihood (CML) method or quantile maximum probability (QMP; Heathcote & Brown, in press). We review the properties of each distribution and the theoretical evidence showing that CML estimates fail for some cases with shifted distributions, whereas QMP estimates do not. In cases in which CML does not fail, a Monte Carlo investigation showed that QMP estimates were usually as good, and in some cases better, than CML estimates. However, the Monte Carlo study also uncovered problems that can occur with both CML and QMP estimates, particularly when samples are small and skew is low, highlighting the difficulties of estimating distributions with parameter-dependent lower bounds. PMID- 15354695 TI - Self-validating presentation and response timing in cognitive paradigms: how and why? AB - With the proliferation of commercial experiment generators and custom software within cognitive psychology and the behavioral sciences, many have assumed that issues regarding millisecond timing accuracy have been largely solved. However, through empirical investigation of a variety of paradigms, we have discovered numerous sources of timing error. These can range from poor scripting practices, to incorrect timing specifications, to hardware variability. Building upon earlier research, we have developed a commercial device and associated software that enables researchers to benchmark most computer-based paradigms in situ and without modification. This gives them the opportunity to correct timing errors where practicable, increase replicability, and reduce variability by altering onset times for stimuli, by replacing inaccurate hardware, or by post hoc statistical manipulation should the source of error be constant. We outline the features of the device and accompanying software suite, stress the importance of such independent validation, and highlight typical areas that can be subject to error. PMID- 15354696 TI - Scientific LogAnalyzer: a web-based tool for analyses of server log files in psychological research. AB - Scientific LogAnalyzer is a platform-independent interactive Web service for the analysis of log files. Scientific LogAnalyzer offers several features not available in other log file analysis tools--for example, organizational criteria and computational algorithms suited to aid behavioral and social scientists. Scientific LogAnalyzer is highly flexible on the input side (unlimited types of log file formats), while strictly keeping a scientific output format. Features include (1) free definition of log file format, (2) searching and marking dependent on any combination of strings (necessary for identifying conditions in experiment data), (3) computation of response times, (4) detection of multiple sessions, (5) speedy analysis of large log files, (6) output in HTML and/or tab delimited form, suitable for import into statistics software, and (7) a module for analyzing and visualizing drop-out. Several methodological features specifically needed in the analysis of data collected in Internet-based experiments have been implemented in the Web-based tool and are described in this article. A regression analysis with data from 44 log file analyses shows that the size of the log file and the domain name lookup are the two main factors determining the duration of an analysis. It is less than a minute for a standard experimental study with a 2 x 2 design, a dozen Web pages, and 48 participants (ca. 800 lines, including data from drop-outs). The current version of Scientific LogAnalyzer is freely available for small log files. Its Web address is http://genpsylab-logcrunsh.unizh.ch/. PMID- 15354697 TI - Reform of statistical inference in psychology: the case of memory & cognition. AB - Geoffrey Loftus, Editor of Memory & Cognition from 1994 to 1997, strongly encouraged presentation of figures with error bars and avoidance of null hypothesis significance testing (NHST). The authors examined 696 Memory & Cognition articles published before, during, and after the Loftus editorship. Use of figures with bars increased to 47% under Loftus's editorship and then declined. Bars were rarely used for interpretation, and NHST remained almost universal. Analysis of 309 articles in other psychology journals confirmed that Loftus's influence was most evident in the articles he accepted for publication, but was otherwise limited. An e-mail survey of authors of papers accepted by Loftus revealed some support for his policy, but allegiance to traditional practices as well. Reform of psychologists' statistical practices would require more than editorial encouragement. PMID- 15354698 TI - A self-novelty manipulation of self-focused attention for internet and laboratory experiments. AB - Conventional manipulations of self-focused attention are poorly suited for Internet experiments and for group-based administration. The authors present a self-novelty manipulation that effectively induces self-awareness for such contexts. In the high self-focus condition, people write about how they differ from their family and friends and from people in general. In the control conditions, people write about neutral topics or do no writing. Three experiments using different measures of self-focus (the situational self-awareness scale, a pronoun selection task, and the private self-consciousness scale) showed that the self-novelty manipulation significantly increased self-focused attention. This effect appeared in Internet-based experiments (Experiment 1) and in laboratory experiments with groups (Experiments 2 and 3). The self-novelty manipulation appears promising for self-awareness research conducted outside of conventional laboratory contexts. PMID- 15354699 TI - ATC-lab: an air traffic control simulator for the laboratory. AB - Air Traffic Control Laboratory Simulator (ATC-lab) is a new low- and medium fidelity task environment that simulates air traffic control. ATC-lab allows the researcher to study human performance of tasks under tightly controlled experimental conditions in a dynamic, spatial environment. The researcher can create standardized air traffic scenarios by manipulating a wide variety of parameters. These include temporal and spatial variables. There are two main versions of ATC-lab. The mediumfidelity simulator provides a simplified version of en route air traffic control, requiring participants to visually search a screen and both recognize and resolve conflicts so that adequate separation is maintained between all aircraft. The low-fidelity simulator presents pairs of aircraft in isolation, controlling the participant's focus of attention, which provides a more systematic measurement of conflict recognition and resolution performance. Preliminary studies have demonstrated that ATC-lab is a flexible tool for applied cognition research. PMID- 15354700 TI - The validity of psychomotor vigilance tasks of less than 10-minute duration. AB - The 10-min psychomotor vigilance task (PVT) has often been used to assess the impact of sleep loss on performance. Due to time constraints, however, regular testing may not be practical in field studies. The aim of the present study was to examine the suitability of tests shorter than 10 min. in duration. Changes in performance across a night of sustained wakefulness were compared during a standard 10-min PVT, the first 5 min of the PVT, and the first 2 min of the PVT. Four performance metrics were assessed: (1) mean reaction time (RT), (2) fastest 10% of RT, (3) lapse percentage, and (4) slowest 10% of RT. Performance during the 10-min PVT significantly deteriorated with increasing wakefulness for all metrics. Performance during the first 5 min and the first 2 min of the PVT deteriorated in a manner similar to that observed for the whole 10-min task, with all metrics except lapse percentage displaying significant impairment across the night. However, the shorter the task sampling time, the less sensitive the test is to sleepiness. Nevertheless, the 5-min PVT may provide a viable alternative to the 10-min PVT for some performance metrics. PMID- 15354701 TI - Using image analysis techniques to analyze figure-copying performance of patients with visuospatial neglect and control groups. AB - Figure copying is often used to detect visuospatial neglect (VSN) in brain damaged patients. We describe algorithms that enable the computation of parameters for describing figure-copying performance. The researcher can readily implement these algorithms on a computer using image analysis software, and they provide information on goodness-of-fit, relative to a standard model, as well as on dynamic aspects of subjects' performance in completing figure copies. To demonstrate the clinical utility of these algorithms, preliminary results from a group of right-hemisphere brain-damaged patients, some of whom have VSN, are compared with those for age-matched controls. PMID- 15354702 TI - Recent advances in operant conditioning technology: a versatile and affordable computerized touchscreen system. AB - We report the construction of a new operant chamber that incorporates modern computer, touchscreen, and display technologies. An LCD display was housed in the front wall of a lightweight Plexiglas chamber. An Apple eMac computer was used to present visual stimuli on the monitor and to control other chamber events. Responses to the stimuli were recorded using a transparent resistive-type touchscreen that overlaid the monitor. The resulting system is simple and inexpensive to construct but powerful and flexible enough to explore a broad range of issues in animal learning and behavior. PMID- 15354703 TI - My perspective on evidence-based practice in dentistry. PMID- 15354704 TI - Human periodontal ligament stem cells isolated for the first time. PMID- 15354705 TI - Teeth first, oral fitness, and the oral theory of everything. PMID- 15354706 TI - Techniques for altering perception of tooth size. PMID- 15354707 TI - Direct composite cavity preparation design and finishing using carbide burs. PMID- 15354708 TI - Treating patients with CARE (comfortable aesthetic restorations): reducing postoperative sensitivity in direct posterior composite restorations. PMID- 15354709 TI - My "high-powereds". PMID- 15354710 TI - Everything I know about endodontic surgery I learned after graduate school. PMID- 15354711 TI - Conservative treatment of the worn dentition with adhesive composite resin. PMID- 15354712 TI - Proper use of Er:YAG lasers and contact sapphire tips when cutting teeth and bone: scientific principles and clinical application. PMID- 15354713 TI - Treatment options for a congenitally missing lateral incisor: a case report. PMID- 15354714 TI - How dentists are judged by patients. PMID- 15354715 TI - The power of e. PMID- 15354716 TI - [Analysis of sports hematuria after running in summer]. AB - BACKGROUND: The recent sport promotion and healthy boom brought increasing the sport population. Running especially is the most popular activity. Various urinary abnormalities are induced by strenunous exercise. Hematuria has been found after various sports activities and noted as sports hematuria by some studies. We evaluated relation between hematuria and running as the most popular sport in summer. MATERIAL AND METHODS: Urine samples were obtained from 109 healthy volunteers participated in 5 km run, at rest before and immediately after running. We could evaluate 90 of all and investigate counts and morphpology of urinary red cells by microscopic finding of urine sediments and analysis of flowcytometry for urinary cells. RESULTS: Eighty-three subjects had increased counts of urinary red cells after running. Of these, 32 had counts above the normal range, microscopic hematuria (RBC above 3/hpf). In analysis of these 32 had dysmorphic pattern mostly. CONCLUSION: This study confirms that exercise induces hematuria easily and sports hematuria has a glomerular source chiefly. PMID- 15354717 TI - [Invasive squamous cell carcinoma of the bladder: report of 18 cases and review of literature]. AB - PURPOSE: This study was undertaken to determine the most effective treatment for improvement of the prognosis of patients with squamous cell carcinoma of the bladder (SCC). MATERIALS AND METHODS: The subjects included 18 cases of invasive SCC (T2 or worse) we have experienced in the past 10 years. While clarifying the clinical patterns of these cases, the association between stage, therapy, and prognosis was studied. Of the cases of invasive SCC reported in Japan in the recent 20 years, 54 cases in which the stage, therapy, and prognosis were documented were selected, and the association between the therapy and outcome in each stage was studied. RESULTS: In our series, 11 cases are alive without cancer for over 2 years. Of the above patients, 7 underwent cystectomy. Cancer death was experienced in 7 patients. Of these patients, 3 underwent cystectomy, and 6 were classified as stage III or higher. As far as our study of the cases reported in Japan is concerned, the prognosis of the cases having undergone TUR or partial resection of the bladder alone was poor. But, even if patients underwent cystectomy, most of the patients was cancer death in the cases whose cancer was stage III or higher. In the patients receiving some supportive therapy, 4 patients receiving radiation plus cisplatin-based chemotherapy were all alive without for over 2 years. CONCLUSIONS: Total cystectomy is most appropriate as the type of operation for the cases of invasive SCC. But, the cases whose cancer was stage III or higher have high recurrence rate, and must be accompanied with some supportive therapy. We concluded that radiation plus cisplatin-based chemotherapy is a candidate of most effective supportive therapy to improve the prognosis of those patients in the supportive therapy. PMID- 15354718 TI - [A case of primary leiomyosarcoma of the spermatic cord]. AB - A 46-year-old man was presented with a firm mass of the spermatic cord. He underwent left radical orchiectomy with clinical diagnosis of spermatic cord tumor. The pathological examination revealed leiomyosarcoma (3.6 x 2.7 x 1.0 cm, pT1bN0M0; 5th TNM, Stage IA; 5th AJCC). He is free of disease 3 year postoperatively. PMID- 15354719 TI - [Acute scrotum from testicular involvement in acute lymphocytic leukemia: a case report]. AB - A 8-year-old boy with acute lymphocytic leukemia (ALL) had received chemotherapy and a complete bone marrow remission was obtained. Then he underwent bone marrow transplantation. After 6 months, he suddenly got left flank-low abdominal pain. Sequentially, he had swelling and redness of left scrotum, left testicular swelling and tenderness. Incision was done and enlarged and hard testis was diagnosed as testicular tumor, left orchiectomy was performed. Histological diagnosis was involvement of ALL, so he received radiotherapy. He remains free of disease by the present after 7 months. PMID- 15354720 TI - [A case of small cell carcinoma of the ureter with SIADH-like symptoms]. AB - Primary small cell carcinoma of the ureter is very rare. We report a case associated with SIADH (syndrome of inappropriate secretion of ADH) -like symptoms. A 53-year-old man presented to our hospital with lower back and left lower quadrant abdominal pain. Computed tomography revealed left hydronephrosis, a peri-ureteral left lower quadrant mass, and retroperitoneal (RP) lymphadenopathy. Transduodendal biopsy of a RP lymph node revealed small cell carcinoma. He was referred to urology for further evaluation. Urography showed an obstructing mass invading the left ureter. Comprehensive metastatic evaluation revealed no other lesions. Thus, we diagnosed primary small cell carcinoma of the ureter with RP lymph node metastases. In addition, he developed SIADH-like symptoms, and we strongly suspected that it was due to ectopic production of ADH from this carcinoma. He was treated with systemic chemotherapy (methotrexate, epirubicin, and cisplatin). Following this, we performed radical nephroureterectomy with RP lymph node resection. However, he died of recurrent disease five months later. PMID- 15354721 TI - [A case of late recurrence of germ cell tumor]. AB - A 19-year-old male was admitted to our hospital because of retroperitoneal tumor and metastases in the lung and liver. He underwent chemotherapy followed by retroperitoneal lymph node dissection and pathologic examination revealed mostly necrotic tissue with a small amount of teratomatous tissue. Two years later, a hyper echoic lesion was found in the left testis, and left high orchiectomy was performed. The resected specimen appeared to be a burned-out testicular tumor. Ten years later, CT scan detected lymph node swelling in the retroperitoneum, and excision of the tumor was performed. Pathologic examination revealed well differentiated adenocarcinoma. Since no primary adenocarcinoma was found, this case was considered late relapse of a germ cell tumor. PMID- 15354722 TI - [A case of advanced rhabdomyosarcoma of the spermatic cord who occurred epilepsy like symptoms, but was completely responded to chemotherapy]. AB - A 19-year-old man presented with left aggressive painless scrotal swelling. On the diagnosis of left intrascrotal tumor, left high orchiectomy with partial scrotal skin resection was performed. Pathological examination of the specimen and systemic metastasis survey revealed embryonal rhabdomyosarcoma of left spermatic cord with multiple lung metastasis (Intergroup Rhabdomyosarcoma Study Group IV). Systemic chemotherapy with etoposide (VP-16), cisplatin (CDDP), and ifosfamide (IFO) (VIP therapy) was started. Although epilepsy-like symptoms occurred at the first course of VIP therapy, these symptoms immediately improved by diazepam administration. These symptoms were thought to be due to the adverse effects of IFO. To our knowledge, there were little reports on epilepsy-like symptoms caused by IFO in Japan. On the other hand, his multiple lung metastasis disappeared after the second course of VIP therapy. Although IFO may be effective in rhabdomyosarcoma, the toxicity of various nervous systems may be discovered. PMID- 15354723 TI - [Diagnostic and prognostic evaluation of unstable angina pectoris using ECG-gated single photon emission computed tomography (SPECT) with 99mTc-tetrofosmin]. AB - PURPOSE: The purpose of this study is to the diagnostic accuracy and the value as prognostic predictors based on the perfusion and/or functional analysis using the QGS program on 99mTc-tetrofosmin gated SPECT (Tf-QGS-SPECT) in patients with unstable angina pectoris (UAP) who admitted to the CCU. METHODS: Fifty seven consecutive patients (37 males and 20 females, mean age: 62 +/- 7 years) with suspected UAP, who underwent Tf-QGS-SPECT and coronary angiography, were entered into this study. We compared the accuracy for diagnosing UAP in the following 3 analyses: 1. Perfusion analysis alone, 2. Functional analysis alone, 3. Perfusion analysis with functional analysis. In addition, we evaluated the prognostic values of the perfusion and/or functional analysis with Tf-QGS-SPECT. RESULTS: Thirty-five of 57 patients with suspected UAP had significant coronary stenosis. The highest accuracy was obtained by adequate combination of perfusion and functional analysis (sensitivity 83%, specificity 82%, accuracy 82%). Thirteen of 29 patients with positive findings in this analysis needed emergent or urgent coronary revascularization. Five of 6 patients with false negative findings in this analysis were left circumflex artery lesions, and two of these patients needed emergent coronary revascularization. CONCLUSION: Evaluation with perfusion and functional images using Tf-QGS-SPECT is useful to improve the accuracy of diagnosis and to predict the prognosis of patients with UAP who admitted to the CCU. PMID- 15354724 TI - [A questionnaire about radiation safety management of the draining-water system at nuclear medicine facilities]. AB - We conducted a questionnaire survey about radiation-safety management condition in Japanese nuclear medicine facilities to make materials of proposition for more reasonable management of medical radioactive waste. We distributed a questionnaire to institutions equipped with Nuclear Medicine facilities. Of 1,125 institutions, 642 institutes (52.8%) returned effective answers. The questionnaire covered the following areas: 1) scale of an institution, 2) presence of enforcement of radiotherapy, 3) system of a tank, 4) size and number of each tank, 5) a form of draining-water system, 6) a displacement in a radioactive rays management area, 7) a measurement method of the concentration of medical radioactive waste in draining water system, 8) planned and used quantity of radioisotopes for medical examination and treatment, 9) an average displacement of hospital for one month. In most institutions, a ratio of dose limitation of radioisotope in draining-water system was less than 1.0, defined as an upper limitation in ordinance. In 499 hospitals without facilities of hospitalization for unsealed radioisotope therapy, 473 hospitals reported that sum of ratios of dose limits in a draining-water system was less than 1.0. It was calculated by used dose of radioisotope and monthly displacement from hospital, on the premise that all used radioisotope entered in the general draining-water system. When a drainage including radioactivity from a controlled area join with that from other area before it flows out of a institution, it may be diluted and its radioactive concentration should be less than its upper limitation defined in the rule. Especially, in all institutions with a monthly displacement of more than 25,000 m3, the sum of ratio of the concentration of each radionuclide to the concentration limit dose calculated by used dose of radioisotope, indicated less than 1.0. PMID- 15354725 TI - [Diagnosis of coronary artery disease by thallium-201 myocardial scintigraphy with intravenous infusion of SUNY4001 (adenosine) in effort angina pectoris--the clinical trial report at multi-center: phase II]. AB - Forty-four patients with effort angina pectoris were evaluated with SUNY4001 (adenosine) thallium-201 (201Tl) myocardial scintigraphy to detect coronary artery disease. These patients had single-vessel disease (> or = AHA 90% stenosis) in either RCA or LAD. Adenosine was infused at the rate of 120 or 140 microg/kg/min for six minutes. 111 MBq of 201Tl was injected after three minutes of the start of the infusion. The early and delayed images were obtained by SPECT imaging. The sensitivity was 94.7% at 120 microg/kg/min and 84.2% at 140 microg/kg/min. Adenosine 201Tl myocardial scintigraphy showed high accuracy for detecting significant coronary artery disease. Adverse reactions occurred in 77.3% of the patients. Regarding the rates of the adverse reactions, there was no significant difference between 120 and 140 microg/kg/min. Major adverse reactions were Chest pain/discomfort (52.3%) and Flushing/Feeling of warmth (27.3%). No serious complication was observed at any infusion rate. Most of adverse reactions disappeared sortly. Only two patients required treatment for moderate chest pain, which, however, disappeared in several minutes. One of the treatments was merely the termination of adenosine infusion, and the other was sublingual spray of nitroglycerin. Adenosine infusion caused slight decrease in blood pressure and increase in heart rate. The hemodynamic changes resolved within several minutes after the adenosine infusion. Decrease in systolic blood pressure of more than 20 mmHg from the base level occurred in 26.1% and 52.4% at 120 and 140 microg/kg/min infusion rate respectively. Therefore, the adenosine infusion at 120 microg/kg/min should be considered safe and useful for the diagnosis of coronary artery disease by pharmacologic stress imaging. PMID- 15354726 TI - [The diagnostic value for ischemic heart disease of thallium-201 myocardial scintigraphy by intravenous infusion of SUNY4001 (adenosine)--the report of clinical trial at multi-center: phase III]. AB - With two hundred and seven patients unable to exercise adequately, the diagnostic accuracy and adverse reaction of 201Tl myocardial scintigraphy with the pharmacologic stress by SUNY4001 (adenosine) infusion were studied. Adenosine was infused for six minutes at the rate of 120 microg/kg/min, and then 201Tl was injected after three minutes from the start of infusion. The early and delayed images were obtained by SPECT imaging. According to angiography, > or = AHA 90% stenosis was defined as significant. The sensitivity of detecting coronary artery disease was 87.1% and the specificity was 46.0%. Adverse reactions occurred in 66.7% of the patients, most of which disappeared shortly with no need for treatment. Major adverse reactions were chest pain/discomfort (30.4%), flushing/feeling of warmth (22.4%) and blood pressure decrease (17.4%). Adenosine infusion caused slight decrease in blood pressure and increase in heart rate. These hemodynamic changes were resolved within several minutes from the termination of adenosine infusion. We concluded that adenosine-201Tl imaging is safe and useful to detect coronary artery disease in patients unable to exercise adequately. PMID- 15354727 TI - [Comparison of myocardial perfusion imaging by thallium-201 single-photon emission computed tomography with SUNY4001 (adenosine) and exercise--crossover clinical trial at multi-center]. AB - We compared the ischemic diagnosis ability and adverse events of 201Tl myocardial perfusion imaging with SUNY4001 (adenosine) stress to that with exercise (ergometer) stress both on random crossover trial. Thirty one known or suspected chronic stable angina patients who are able to exercise and 10 healthy volunteers were enrolled for the trial. The early and delayed images were obtained by SPECT imaging. The concordance of diagnoses [ischemia vs. no ischemia] between the two types of stresses was 97.3% (36/37) [Kappa: 0.9068]. The sensitivity and specificity based on the exercise test were 100% (6/6) and 96.8% (30/31) respectively. The incidence of adverse events caused by SUNY4001 and the exercise were 44.7% (17/38) and 52.6% (20/38), respectively. Major adverse events caused by SUNY4001 were BP decrease, flushing and headache. And those by exercise were ST decrease, dyspnea and chest pain. None of the adverse events required the intervention or caused life-threatening complication in the trial. The trial showed that the ischemic diagnosis ability and safety of 201Tl scintigraphy with SUNY4001 stress are almost equal to those of the exercise stress that is considered as the standard stress method. We concluded that 201Tl imaging with SUNY4001 is safe and useful for detecting ischemic heart disease, especially for patients unable to exercise adequately. PMID- 15354728 TI - [Clinical evaluation on isolation of Mycobacterium kansasii in our hospital]. AB - MATERIALS AND METHODS: We retrospectively evaluated 49 cases from whom Mycobacterium kansasii (MK) was isolated from 1992 to 2001 in our hospital. RESULTS: The annual numbers of MK patients have increased. One of the clinical characteristics of patients in our cases was relatively low rate of cavitary lesions. In 13 patients who had not fulfilled the diagnostic criteria of MK infection, the clinical disease due to MK did not appear at all during observation period ranging one month to ten years. CONCLUSION: These findings suggest that MK isolation from clinical specimens is not always considered clinically significant, but may be colonization. PMID- 15354729 TI - [Mycobacterium shinshuense isolated from cutaneous ulcer lesion of right lower extremity in a 37-year-old woman]. AB - PURPOSE: Second clinical infection case of Mycobacterium shinshuense was presented, we tried the identification of M. shinshuense that is isolated from skin. OBJECT: Mycobacteria species isolated from cutaneous ulcer lesion of right lower extremity in a 37-year-old woman. METHOD: Identification by DNA-DNA Hybridization, 16S rRNA and rpoB method as genomic level and conventional method. RESULT: It did not grow on 1% Ogawa's slant medium at both 37 degrees C and 42 degrees C, but grew at 28 degrees C. It formed yellowish colonies in the dark. It was difficult to distinguish M. shinshuense from M. ulcerans and M. marinum by DNA-DNA hybridization (DDH) and DNA sequencing. To identify that it is M. shinshuense, growth rate, temperature range of mycobacterial growth, light coloration reaction, biochemical and biological tests, and drug susceptibility testing were further explored. Finally it was identified as M. shinshuense based on these CONSIDERATION: For Mycobacteria species which grow 2 weeks after inoculation at 28 degrees C, and which is identified as M. marinum by DDH method, it is necessary to identify with sequence and conventional method. PMID- 15354730 TI - [Evaluation of quantitative fit-testing of N95 filtering facepiece respirators using Mask-Fitting Tester and improvement of mask fitting by instruction]. AB - OBJECTIVE: To evaluate quantitative fitness of N95 filtering facepiece respirators (N95 Mask) using Mask Fitting Tester and improvement of fitness by instruction. MATERIALS AND METHODS: One hundred and thirty-three health care workers working at our hospital were tested as to quantitative fitness of N95 Mask using Mask Fitting Tester Model MT-02, Roken type that counts particles more than 0.7 microm in diameter. Based on counting the number of particles more than 0.7 microm in diameter of the air inside and outside the mask, leakage rate is calculated. Permissible range of leakage rate is 10% or below. In case of leakage rate more than 10%, we instructed way of wearing N95 Mask or change a type of N95 Mask. We usually provide three types of N95 Mask. RESULTS: Eighty-seven (65.4%) out of 133 health care workers achieved leakage rate 10% or below at the first test. Forty-six (34.6%) who did not achieve leakage rate 10% or below at the first test received instruction or changed a mask type. Twenty-one (15.8%) achieved desired value by instruction and 19 (14.3%) by changing a mask type. But 6 (4.5 %) could not achieve desired value in spite of instruction or changing to another type of masks available to us. Leakage rate changed from 20.5 +/- 10.9% (10.1-58.6) to 6.3 +/- 4.9% (0-29.5) by instruction or changing a type of mask (p < 0.001). At the first test, mask fitness is better in male than female, and worse in workers who always use N95 Mask at isolation ward or laboratory room than workers who usually do not use N95 Mask. Main problems were choice of mask, fitness to nose, incorrect use of headband. CONCLUSION: It is useful to evaluate N95 Mask quantitative fitness using Mask Fitting Tester Model MT-02, Roken type. At the first test, 34.6% of health care workers have large amount of leakage more than 10%, and by instruction or changing a mask type, most of them could achieve 10% or below. But 4.5% could not achieve desired value. On Infection control, it is important to test quantitative fitness of N95 Mask and to provide more than three types of N95 Mask. PMID- 15354731 TI - [Pseudo-recurrence of lung tuberculosis based on the detection of smear AFB positive sputum due to excretion of necrotic material]. AB - A 21-year-old man was admitted to our hospital for cough and hemoptysis. The patient showed smear positive pan-sensitive lung tuberculosis, and completed standard course of chemotherapy successfully. Six months after the completion of chemotherapy he had hemoptysis again. The chest radiograph showed that pre existing tuberculoma-like shadow in the right upper lobe was changed to a cavity. Although sputum smear examinations revealed positive results several times, sputum culture was always negative. This clinical exacerbation was thought to be "pseudo-recurrence" due to excretion of necrotic material from the pre-existed abscess nodule. His disease improved without any anti-tuberculosis chemotherapy. Diagnosis of lung tuberculosis recurrence should be made on sputum culture positive results. PMID- 15354732 TI - [A case of cervical and mediastinal lymph nodes tuberculosis, tuberculous pleurisy, spinal caries and cold abscess in the anterior chest wall]. AB - A 61-year-old woman with schizophrenia that had been treated in a psychiatric hospital was admitted to our hospital because of subileus and back pain. Though subileus was improved, she had a sudden attack of fever 7 days later and developed right pleural effusion, a cold abscess in the anterior chest wall and swelling of a thumb-sized right cervical lymph node which broke through the skin. We made a diagnosis of cervical and mediastinal lymph nodes tuberculosis, tuberculous pleurisy, spinal caries and cold abscess in the anterior chest wall due to the biopsy findings of the specimen taken from the cervical lymph node, examination of pleural effusion, chest CT, bacteriological examination of the cold abscess and spinal MRI. We started chemotherapy with the antituberculous drugs (HRSZ) and symptoms except back pain improved. She complained of paresis of the both lower extremities, which completely paralyzed 8 months later in spite of continued chemotherapy. Thereafter her paralysis was gradually improved and she was able to walk by herself after 12 months chemotherapy. PMID- 15354733 TI - Immunophenotypic and genotypic characterization of progression in follicular lymphomas. AB - Progression of follicular lymphomas (FLs) is often accompanied by a spectrum of histologic changes and an aggressive clinical course. Although molecular alterations have been implicated in this event, the underlying factors are largely unknown. We studied the expression of selected tumor suppressor genes (P53 and retinoblastoma [RB]), oncogenes (MYC and BCL2), and a transferrin receptor related protein (Trump) in sequential biopsies in 16 patients. Eleven patients progressed from grade I or II FL to aggressive B-cell lymphomas with diffuse morphology, whereas 5 patients presented with diffuse aggressive lymphomas and recurred with indolent lymphomas. Immunoreactivity for P53 correlated with higher histologic grade in lymphomas progressing from indolent to aggressive; however, only 1 patient who presented with aggressive lymphoma demonstrated a P53 gene mutation. Neither P53 immunoreactivity nor genotypic alterations correlated with presentation with an aggressive histology and relapse with FL. Growth fraction, as assessed by Ki-67 staining, and Trump expression correlated with histologic grade. Immunoreactivity for RB, BCL2, and MYC was seldom associated with progression. Eight of 9 cases tested exhibited identical immunoglobulin heavy and light chain rearrangements or identical BCL2 gene rearrangements in the sequential lymphomas. We conclude that P53 and Trump protein expression and proliferation activity correlate with histologic grade, but not with recurrence or progression of FL. Our results further indicate that progression of FL to diffuse aggressive lymphomas and presentation of an aggressive B-cell lymphoma followed by FL are clonally related. PMID- 15354734 TI - Expression of c-FLIP in classic and nodular lymphocyte-predominant Hodgkin lymphoma. AB - Different molecular pathways are believed to be involved in the pathogenesis of classic Hodgkin lymphoma as opposed to non-Hodgkin lymphoma. Antiapoptotic mechanisms have been proposed for classic Hodgkin lymphoma, including expression of the cellular Fas-associated death domain-like interleukin-1beta-converting enzyme inhibitory protein (c-FLIP), which plays a critical role in resistance to CD95/Fas-mediated apoptosis. In this study, we compare the expression of c-FLIP in the neoplastic cells of classic Hodgkin lymphoma and nodular lymphocyte predominant Hodgkin lymphoma cases. Sixteen cases of classic Hodgkin lymphoma and 19 cases of nodular lymphocyte-predominant Hodgkin lymphoma were reviewed. Of 16 classic Hodgkin lymphoma cases, 13 cases (81%) were c-FLIP-positive, compared with 6 of 19 (32%) nodular lymphocyte-predominant Hodgkin lymphoma cases. Strong cytoplasmic staining was seen in 7 of 13 c-FLIP-positive classic Hodgkin lymphoma cases, in contrast with only 2 of 6 c-FLIP-positive nodular lymphocyte predominant Hodgkin lymphoma cases. The 2 cases of nodular lymphocyte-predominant Hodgkin lymphoma with strong c-FLIP expression were associated with transformation to large B-cell lymphoma. An additional 15 cases of diffuse large B-cell lymphoma were studied for c-FLIP expression. All but 1 were c-FLIP positive. In conclusion, we detected c-FLIP in a significantly lower proportion of nodular lymphocyte-predominant Hodgkin lymphoma cases compared with classic Hodgkin lymphoma cases. Therefore, c-FLIP expression may not be the major mechanism of pathogenesis in nodular lymphocyte-predominant Hodgkin lymphoma. However, strong c-FLIP expression in nodular lymphocyte-predominant Hodgkin lymphoma was associated with transformation to large B-cell lymphoma in 2 cases. c-FLIP expression is not limited to Hodgkin lymphoma, because the majority of diffuse large B-cell lymphoma cases tested were strongly c-FLIP-positive. PMID- 15354735 TI - Expression profiling of colorectal carcinomas using tissue microarrays: cell cycle regulatory proteins p21, p27, and p53 as immunohistochemical prognostic markers in univariate and multivariate analysis. AB - With the rapidly growing understanding of tumor biology, molecular staging of cancer is expected to improve prognostication. This would be particularly important for cancers amenable to adjuvant treatment, such as colorectal carcinomas. To generate data for this, the tissue microarray technique may prove useful. Tissue microarrays were constructed with triplicate cores (0.6 mm diameter) from the invasive margins of a consecutive single-institution series of 184 colorectal carcinomas. Immunostaining for p53, p21, p27, Ecadherin, and beta catenin was scored. Tumor cell proliferation was assessed by mitotic indices and Ki-67 labeling, apoptosis by quantification of apoptotic bodies. Reduced nuclear immunostaining for p21 (<10%) and p27 (< or =50%) and reduced membranous expression of Ecadherin were significantly associated with a poorer clinical course by univariate analysis. beta-catenin immunostaining had no prognostic impact. Mitotic and apoptotic indices as well as Ki-67 labeling below the median were indicators of poor prognosis. Complete absence of p53 nuclear staining was a significant adverse prognostic factor. By Cox regression, p53 = 0%, p53 = 0%, in combination with p27 < or = 50%, the mitotic index and the combined mitotic and apoptotic index added prognostic information to UICC stage. The authors found that growth pattern, lymphohistiocytic response, lymphatic permeation, and venous spread, too, each was a strong prognosticator in addition to UICC stage. The results support that tissue microarrays are a useful tool for screening immunohistochemical markers for prognostic use. An immunopanel of p21, p27, and p53 could be useful for prognostication in colorectal carcinoma in addition to UICC stage. PMID- 15354736 TI - Wilms tumor gene product: sensitive and contextually specific marker of serous carcinomas of ovarian surface epithelial origin. AB - Carcinomas of ovarian surface epithelial origin can arise from, and often present at, extraovarian sites. There are few available markers for the positive identification of carcinomas of ovarian surface epithelial origin, which might aid in distinguishing them from metastatic carcinomas, such as of breast, colon, or lung origin. Recently, the Wilms tumor gene product (WT-1) has been shown to be expressed in ovarian surface and mesothelial epithelium. We tested the hypothesis that WT-1 would be a sensitive and specific marker of ovarian surface epithelium carcinomas. An archived series of 116 ovarian carcinomas (57 serous [43 ovarian, 14 extraovarian], 31 mucinous, 15 clear cell, 13 endometrioid), 118 breast carcinomas, 46 colonic carcinomas, and 45 nonsmall cell lung cancers were selected. A polyclonal antibody to the WT-1 gene product was applied to deparaffinized, formalin-fixed tissue sections after epitope retrieval. Fifty-two of 57 (93%) serous carcinomas of ovarian surface epithelial origin were WT-1 positive, in a nuclear pattern, with virtually all the tumor cell population positive in the majority of cases. None of the mucinous, clear cell, or endometrioid ovarian cancers were positive, and only 8 of 118 breast, 0 of 46 colonic, and 0 of 45 lung nonsmall cell carcinomas were WT-1-positive. These findings demonstrate that WT-1 is a highly sensitive and specific marker of serous carcinomas of ovarian surface epithelial origin (both ovarian and extraovarian). These results also contradict recent reports demonstrating WT-1 expression in both breast and lung carcinomas. PMID- 15354737 TI - Utility of CDX-2 in distinguishing between primary and secondary (intestinal) mucinous ovarian carcinoma: an immunohistochemical comparison of 43 cases. AB - Primary and secondary mucinous tumors can involve the ovaries and have similar histologic appearances. The differential diagnosis is important for surgical and chemotherapeutic treatment and for the prognosis, but often it is extremely difficult. This article discusses an immunohistochemical panel that includes carcinoembryonic antigen (CEA), cytokeratin (CK) 7, CK20, CA125, CA19.9, and a new marker, CDX-2, for the distinction between primary ovarian mucinous carcinomas and metastatic (intestinal) ovarian tumors. Forty-three cases representing primary and secondary ovarian tumors were considered and consisted of 14 primary mucinous ovarian carcinomas (PMOCs) and 29 secondary (intestinal) ovarian tumors (SIOTs). Fisher exact test was performed to evaluate the reliability of the respective antibodies to discriminate between PMOCs and SIOTs. CDX-2 was diffusely positive in all SIOTs and was expressed focally in 3 cases (21.42%) of PMOCs. CK7 was diffusely positive in 13 cases (44.82%) of SIOTs and in 13 cases (92.85%) of PMOCs. CK20 was diffusely positive in 17 cases (58.62%) of SIOTs and in 6 cases (42.85%) of PMOCs. CEA was diffusely positive in 28 cases (96.55%) of SlOTs and in 12 cases (85.71%) of PMOCs. CA 19.9 was positive in all SIOTs and in 12 cases (85.71%) of PMOCs. CA125 was positive in 3 cases (10.34%) of SIOTs and in 4 cases (28.57%) of PMOCs. CK7 and especially CDX-2, a specific and sensitive marker, can aid pathologists in making a differential diagnosis (P = 0.003 and P < 0.0005, respectively), whereas CEA, CK20, CA125, and CA 19.9 markers are not high enough to distinguish between primary and secondary mucinous ovarian tumors. PMID- 15354738 TI - Expression and localization of GRP75 in human epithelial tumors and normal tissues. AB - Using differential display mRNA techniques, the authors found cDNA of the heat shock 70 protein known as GRP75 overexpressed in ovarian cancer cell lines. In the current study, the authors used immunohistochemistry to characterize the expression pattern of GRP75 in ovarian carcinomas and compared it with epithelial tumors originating from the female reproductive tract, epithelial neoplasms from non-gynecologic sites (colon, pancreas, breast, and lung), and various normal tissues. The authors also developed an antigen capture ELISA assay to determine if GRP75 can be detected in tumors, ascites, or sera of patients with advanced mullerian adenocarcinomas. All epithelial tumors from the ovary and the female reproductive tract were positive for GRP75 expression with moderate to strong staining intensity; stromal expression of GRP75 was generally weak or absent. Adenocarcinomas from the colon, lung, pancreas, and breast also stained strongly positive for GRP75. The epithelial cells of all normal tissues examined were positive for GRP75, and strong staining was also seen in the corpora lutea, hepatocytes, enteric neural plexus of the esophagus and colon, and placental cytotrophoblast and syncytiotrophoblast, and in subpopulations of pancreatic acinar cells. The ELISA assay detected GRP75 in tumor lysates and ascitic fluid, but not sera, of patients with mullerian adenocarcinomas. The authors conclude that GRP75 is highly expressed in both benign and malignant epithelium, as well as cells of specialized function from a variety of tissues. PMID- 15354739 TI - Rates of estrogen receptor-alpha expression are no different in low-risk (Vietnam) and high-risk (Australian) breast cancer. AB - Past studies have led to the view that positive estrogen receptor-alpha (ERa) expression is present in a 20% higher proportion of breast cancer in postmenopausal, but not premenopausal, women from populations with a high risk of breast cancer such the United States compared with populations with a low risk of breast cancer such as Japan. Data from the early studies that compared ERa expression in breast cancer specimens from Japanese and US women may not be reliable because no account was made of differing methods of laboratory analysis, grade (which has a profound influence on ERa expression), type of breast cancer, and threshold value (the cutoff level of Era-positive cells that defines positivity or negativity). We hypothesized that if these variables were standardized, there would be little or no difference in ERa expression in breast cancer in women from populations at low and high risk of breast cancer. Immunohistochemical analysis of ERa was conducted on formalin-fixed, paraffin mounted, invasive breast cancer biopsy specimens from 107 Vietnamese (Hanoi) and 201 Australian subjects, which has enabled us to make valid comparisons of ERa expression between women from populations at very low and very high risk of breast cancer. There was no statistically significant difference in the percentage of ERa-positive breast cancer specimens between Vietnamese and Australian subjects for each 5-year age group, except the age group 51 to 55 years. For this age group, the percentage of ERa-positive specimens was significantly higher among the Vietnamese specimens. Our research question was, "When the known variables influencing ERa expression in breast cancer are standardized, is there any difference in ERa expression in breast cancer in women from populations at low and high risk for breast cancer?" The answer is, with respect to Vietnam, no. We have observed that ERa expression in breast cancer among Vietnamese women is higher than previously believed. It is possible that ERa expression in breast cancer among women from other developing countries is also higher than previously believed. The implication is that estrogen modulators such as tamoxifen, which are effective in the treatment of ERa-positive breast cancer, probably have a substantial role in developing and developed countries. PMID- 15354740 TI - Cytokeratin 7/20 and MUC1, 2, 5AC, and 6 expression patterns in Barrett's esophagus and intestinal metaplasia of the stomach: intestinal metaplasia of the cardia is related to Barrett's esophagus. AB - Intestinal metaplasia (IM) in endoscopic biopsies obtained from close to the gastroesophageal junction may represent IM of the cardia (CIM) or Barrett's esophagus (BE), which have different malignant potentials despite similar morphology. This study compared cytokeratin (CK) 7/20 and mucin (MUC1, 2, 5AC, and 6) immunopatterns in biopsies from BE (n = 41), CIM (n = 35), and antral gastric IM (AIM, n = 37) to evaluate their roles as diagnostic aids. CK7 and CK20 expression was described as absent, patchy (superficial and deep), continuous superficial only, continuous deep only, and diffuse. Eleven different combinations of CK7/20 expression were seen. Since CK20 staining was positive in all cases, four main patterns were defined on the basis of the observed CK7 staining as 1, absent; 2, patchy (superficial and/or deep); 3, diffuse; and 4, continuous superficial only. Overall CK7 positivity (regardless of pattern) was higher in BE and CIM than in AIM. CK patterns 3 and 4 were also higher in BE and CIM than in AIM. For either pattern 3 or 4, the positive and negative predictive values for BE versus AIM were 95% and 67%, respectively. MUC1 was rarely expressed in BE and CIM compared with AIM, whereas the opposite was noted for MUC5AC expression. MUC2 and MUC6 expression was similar in all locations. In conclusion, diffuse or continuous superficial CK7 staining is highly characteristic of BE and CIM and contrasts with AIM. It is, however, not very sensitive. CK20 profiles have no added value. Mucin expression also differs between BE and CIM versus AIM, but the specificity of any pattern is insufficient for distinction in individual cases. Importantly, CK and MUC expression patterns in BE and CIM are virtually indistinguishable, limiting their use in this differential and raising the question of whether they are biologically related. PMID- 15354741 TI - Inhibin expression in ovarian-type stroma in mucinous cystic neoplasms of the pancreas. AB - Mucinous cystic neoplasms (MCNs) of the pancreas are typically found in middle aged to elderly women and contain ovarian-type stroma in the cyst wall. Whether the resemblance of this stroma to ovarian stroma is only morphologic or has more functional similarity is still unclear. Estrogen receptors (ER) and progesterone receptors (PR) have been shown to be expressed in a wide variety of tissues and tumors, including the ovarian-type stroma of MCN. Inhibin, on the other hand, has been shown to have a more restricted expression, limited to ovarian sex cord stromal components and placental cells, and has recently been shown to be expressed in pancreatic MCNs. However, it is still unclear whether this expression is limited to MCNs of the pancreas and whether it has any diagnostic role. Seven cases of MCN (4 mucinous cystadenoma, 2 borderline MCN, and 1 mucinous cystadenocarcinoma with microinvasion), 6 cases of intraductal papillary mucinous tumor, 1 of mucinous cystic tumor of uncertain classification, 2 of mucinous noncystic adenocarcinoma, 4 of serous cystadenoma, and 4 solid pseudopapillary neoplasms were selected for this study. Five cases with normal pancreatic tissue were included as controls. Immunohistochemical stains for alpha inhibin, ER, and PR were performed on a representative section from each case on formalin-fixed, paraffin-embedded tissue sections using a standard indirect immunoperoxidase method. All cases of MCN were in female patients with an average age of 55.3 years, showing ovarian-type stroma and clusters of alpha-inhibin positive luteinized theca-like cells. In all these cases, moderate to strong PR positivity was also noted in the ovarian-type stroma, including many of the alpha inhibin-positive luteinized theca-like cells. ER was expressed in 2 cases. The epithelial cells of MCNs were all negative for ER, PR, and alpha-inhibin staining. Of the other tumors, 4 solid pseudopapillary neoplasms showed positivity for only PR in the tumor cells. The remaining tumors were negative for all markers. In conclusion, the finding of alpha-inhibin positivity in MCN with ovarian-type stroma further supports its similarity to true ovarian stromal tissue and may suggest a role of complex hormonal interaction in the pathogenesis. In addition, its limited expression in MCNs of the pancreas may be diagnostically useful in difficult cases. PMID- 15354742 TI - Immunohistochemical detection of P504S in primary and metastatic renal cell carcinomas. AB - P504S/alpha-methylacryl CoA racemase has been shown to be a relatively sensitive and specific positive marker for prostatic adenocarcinoma. The potential utility of P504S in renal cell neoplasms has not been explored in a large series. We assessed the diagnostic value of P504S in 332 cases of nonprostatic neoplasms using the avidin-biotin-peroxidase complex technique, including 115 renal neoplasms, 28 metastatic renal cell carcinomas (RCCs), and 189 nonrenal neoplasms. The results demonstrated that a granular, cytoplasmic staining pattern for P504S was observed in 48 of 70 (68.6%) conventional (clear cell) RCCs, 15 of 15 (100%) papillary RCCs, 2 of 7 (29%) chromophobe RCCs, and 2 of 8 (25%) oncocytomas. Among the 70 cases of clear cell RCC, positivity of P504S was seen in 40%, 71%, 94%, and 75% of RCCs with Furhman nuclear grade I, II, III, and IV, respectively. Strong immunostaining was present in each case (86/86) in the proximal tubules adjacent to the renal neoplasm. Eighty-two percent of metastatic RCCs (23/28) were positive for P504S. However, only 24 of 189 (13%) nonrenal malignancies were positive. The 24 positive cases included 12 of 13 (92%) colorectal adenocarcinomas, 6 of 30 (20%) ductal carcinomas of the breast, and 4 of 23 (17%) adenocarcinomas of the lung. These findings suggest that P504S is a useful marker in diagnosing primary and metastatic RCCs, although it has little value in differentiating chromophobe RCC from oncocytoma. PMID- 15354743 TI - Ewing sarcoma/peripheral primitive neuroectodermal tumor: adult abdominal tumors with an Ewing sarcoma gene rearrangement demonstrated by fluorescence in situ hybridization in paraffin sections. AB - The differential diagnosis of small round cell tumors is exhaustive and requires ancillary studies. Relatively recently, fluorescence in situ hybridization (FISH) using probes for specific gene rearrangements has gained wide acceptance. This technique is particularly useful in the differential diagnosis of Ewing sarcoma/primitive neuroectodermal tumor (ES/PNET) and desmoplastic small round cell tumor (DSRCT). In ES/PNET, the EWS gene is juxtaposed to the FLI-1 gene in 85% of cases and to the ERG gene in another 7% of cases; the EWS gene is juxtaposed to the WTI gene in DSRCT. Documentation of the EWS gene rearrangements in EWS/PNET has previously been demonstrated in frozen tissue. We report 2 unusual cases of EWS/PNET diagnosed in abdominal tumors in adults. Although the immunohistochemical results supported a diagnosis of ES/PNET, 1 case morphologically resembled DSRCT. The diagnosis in these 2 cases was confirmed by the FISH demonstration of EWS/FLI-1 gene fusion in paraffin-embedded tissue. Thus, the usefulness of FISH demonstration of an EWS gene rearrangement with these specific probes in such unusual cases is supported and is demonstrated in paraffin-embedded tissue. PMID- 15354744 TI - Immunoperoxidase staining for C4d on paraffin-embedded tissue in cardiac allograft endomyocardial biopsies: comparison to frozen tissue immunofluorescence. AB - C4d deposition in microvasculature is a marker for humoral rejection. The authors compared a recently developed C4d immunoperoxidase (IP) method for paraffin embedded tissue to immunofluorescence (IF) of frozen tissue. Of 315 frozen endomyocardial biopsies with IF staining for C4d, 280 were negative and 35 were positive. Negative controls were 17 negative biopsies and 11 biopsies with myocyte necrosis. The extent of IP and IF staining was graded as 0 to 3+. Staining intensity and the number and type of positive vessels were recorded. Staining patterns in Quilty lesions (QL) and foci of acute cellular rejection (ACR) were also evaluated. In 34 biopsies with sufficient tissue, IP criteria of 2+/3+, or more than 10 to 20 positive vessels per 10 high-power fields detected 25.0% (1/4), 18.2% (2/11), and 84.2% (16/19) of 1+, 2+, and 3+ IF-positive biopsies, respectively, without false positives. Considering C4d IF 3+ as positive resulted in 84.2% (16/19) sensitivity and 93.0% specificity (40/43). Intensely stained capillaries predominated in six of seven biopsies when more than 100 capillaries per 10 high-power fields were positive. Seventy percent (7/10) of IP 2+ and 3+ biopsies showed positive capillaries in QLs, while 36.4% (4/11) of IP 1+ and negative biopsies did. All eight IP 2+/3+ biopsies showed positive capillaries in ACR foci, while 25.0% (1/4) of IP-negative biopsies did. Capillary staining in QLs and areas of ACR reflects overall C4d deposition. In conclusion, IP staining of 2+/3+ is highly sensitive and specific for C4d positivity. The authors recommend considering 2+ and 3+ as positive staining when using the IP technique. PMID- 15354745 TI - Can a more selective application of antigen retrieval facilitate standardization in immunohistochemistry? AB - A clearer perception of the major immunochemical forces participating in the reaction between tissue antigens and their complementary antibodies would help us control better the processes of formalin fixation and antigen retrieval. This report describes and discusses the results of additional experiments designed for this purpose. It is anticipated that they will make a useful contribution to the much needed standardization of staining in immunohistochemistry. PMID- 15354746 TI - Comparison of immunoexpression of 2 antibodies for estrogen receptors (1D5 and 6F11) in breast carcinomas using different antigen retrieval and detection methods. AB - The importance of in situ immunodetection of hormone receptors for therapy planning and prognostic evaluation in patients with breast carcinoma is well established. Sensitive detection methods are of utmost importance, especially in poorly fixed tissues, which are not uncommon in routine pathologic practice. The purpose of the present study is to compare immunoexpression of estrogen receptors in 20 cases of invasive ductal carcinoma using two antibodies, 1D5 and 6F11, and to verify the effect of different antigen retrieval solutions and detection systems. Immunoperoxidase was performed on paraffin sections using 1D5 and 6F11 as primary antibodies. Heat-induced antigen retrieval was performed using citrate buffer (pH 6.0) or Tris-EDTA buffer (pH 8.9). Detection was achieved using the following systems: EnVision, EnVision Plus, and labeled streptavidin-biotin peroxidase complex. Reaction was semiquantified from 0 to 4. There were no differences between the two markers, 1D5 and 6F11, except when 6F11 was used with EnVision and citrate buffer, in which case weaker reactivity was observed. Only in this combination (6F11/EnVision) was EDTA buffer significantly better than citrate. Labeled streptavidin-biotin peroxidase complex presented the best results, followed by EnVision Plus. PMID- 15354747 TI - [The foot -- our youngest skeletal segment]. PMID- 15354748 TI - [Hallux valgus--an atavism?]. AB - In biology, atavism is generally understood as a biological phenomenon which brings to sight a recurrent phenotypic character which was lost for extinct generations. Phylogenic evolution of homo sapiens demonstrates today that the first plantar ray in tetrapods has always been a very stable ray and the lateral foot experienced a progressive adduction to the talo-metatarsal axis, together with a remodeling of the lateral column of the foot and a progressive valgus of the calcaneus beneath the talus. The foot appears as a strongly modified extremity, starting at a prehensile foot organ, common to our ancestors. In homo sapiens, the hand became a very mobile, less loaded organ, while the foot became a rather less mobile, but more loaded organ. The first ray, distal to Lisfranc's ligament, has no ligamentous structure holding it to the second ray. Stability is thus precarious and controlled by extrinsic and intrinsic muscles. Passive mobility of the tarso-metatarsal joints has been studied in vitro and in vivo. A relationship between unstable or hypermobile first ray and relevant Hallux valgus deformity has been demonstrated. Factors which facilitate the deformity are a long hallux and a functional equinus of the foot. Clinical pathology includes metatarsalgia, hammer toes, together with metatarsus primus varus and shortened gastrocnemii. Logical treatment of relevant and painful deformity includes tarso metatarsal fusion with or without fusion to the second metatarsus. Osteotomies seem less secure in severe cases on the long run. In conclusion, Hallux valgus cannot be considered as an atavism because clinical history and findings do not correspond to the archaic prehensile foot and its evolution, but rather do correspond to a weak spot on a (still) fragile, often overloaded, phylogenetically young organ. PMID- 15354749 TI - ["Hallux rigidus"--a global problem?]. AB - The diagnosis "hallux rigidus" is used for limitation of great toe motion, which is particularly painful at push-off during gait. The underlying condition is often degenerative arthrosis of the first metatarsophalangeal joint. Radiographs show no abnormality initially, later degeneration sets in at the dorsal aspect of the joint. Conservative treatment mainly consists of local anti-inflammatory applications and orthopaedic appliances to decrease load at the metatarsophalangeal joint. With progression of the arthrosis, joint preserving procedures such as cheilectomy are used. With this technique, the degenerated dorsal part of the first metatarsal head is resected. In complete destruction of the joint space, arthrodesis is the technique of choice in the mostly active, younger patients. Resection arthroplasty is a simple technique which is used in elderly, less active patients. The results of first metatarsophalangeal joint replacement are inferior to arthrodesis. PMID- 15354750 TI - [Lesser toe deformities]. AB - Lesser toe deformities often lead to painful calluses and metatarsalgia. Depending on the different underlying etiologies it is mandatory to perform a meticulous clinical assessment including the whole foot and the entire lower limb. Prior to any surgical interventions it is necessary to evaluate the deformity at all three joint levels. The metatarsophalangeal joint acts as a key joint. Any dorsal subluxation or dislocation has to be addressed first. This may include various soft tissue procedures and shortening osteotomies of the metatarsals. After successful realignment contractures of the distal joints have to be corrected. Since function of the lesser toes mainly depends on stability of the distal joints arthrodeses of the proximal and distal interphalangeal joints are superior to any resection arthroplasties. PMID- 15354751 TI - [Diabetic foot disease--a review of pathogenesis, treatment and prevention of diabetic podopathy]. AB - An update is presented of the pathogenesis, treatment and prevention of diabetic podopathy. Although the underlying conditions, polyneuropathy and peripheral ischaemic vessel disease presently cannot be cured, their complications, i.e. foot ulcers, foot gangrene and foot fractures in most cases can nowadays be treated successfully without major amputations. PMID- 15354752 TI - [Talus fractures--fractures of the most important tarsal bone]. AB - The talus is the key bone of the foot due to its location between the ankle and the subtalar joints. Through the flexion and extension of the ankle joint, the talus is the "propulsive bone" situated at the root of the first ray and the hallux. Through the subtalar and talo-navicular joints, the talus allows the foot to be "suspended" using a fancy spring mechanism involving ligaments and tendons. The talus thus transmits forces through two important joints without any direct muscular constraint. The talo-calcaneo-navicular joint may be called the "coxa pedis" due to the anatomical and functional similitudes with the coxofemoral joint ("coxa pelvis"). Fractures of the talus are rare. Talus fractures can be classified in "central" and "peripheral" fractures. Central fractures occur through a strong axial blow provided that the ankle and coxa pedis are locked by extrinsic muscular contraction. Peripheral fractures instead occur in an extreme position of the ankle or the coxa pedis, with a subluxation or complete dislocation of one or more of those joints. Central fractures produce a bony solution of continuity in between at least two of the ankle, subtalar and talo navicular joints. Peripheral fractures mostly do implement joint surfaces which are sheared off. Aim of treating talus fractures is precise and stable reduction of the fragments because most often, the fractures cross the articular layers. The surgical approaches are critical, especially in displaced central fractures, because the reduction may require visual control all around the bone. There are three approaches which might be used as single approaches or combined simultaneously: the anteromedial approach along the subtalar joint from the navicular to the retro-malleolar region, the anterolateral approach centered on the sinus tarsi (Ollier) and the postero-lateral approach (Gallie). In many cases, a joint distraction device may help visualization of the different joint spaces as well as assist reduction of dislocations. Early joint mobilization, without weight bearing may be very profitable for good functional results. Aseptic necrosis (AVN) of the talus do not need special care if asymptomatic. On the long run, open operative arthrolysis might be added to the eventual removal of implants to considerably improve the results. PMID- 15354753 TI - [Calcaneal fractures--the most frequent tarsal fractures]. AB - Operative treatment of intra-articular calcaneal fractures with anatomical reduction and early mobilisation has lead to a considerable improvement of the functional results of these severe injuries over the recent two decades. New tendencies like modified approaches tailored according to the vascular anatomy, arthroscopic control of joint reduction in conjunction with open or percutaneous procedures, early flap coverage for severe open fractures or closed fractures with full thickness skin necrosis, the use of interlocking plates that further minimises the need of bone grafting and subtalar arthrolysis at the time of hardware removal for fibrous overgrowth have the potential for further improvement of the outcome. The observation from several comparative studies that operative treatment leads to superior results in displaced intra-articular calcaneal fractures has been qualified in a recent prospective, randomised multicentre trial. This paper presents and the authors' experience with the demanding operative treatment of these fractures together with new developments aiming at improving the clinical results. PMID- 15354754 TI - [Nutcracker fractures of the navicular and cuboid]. AB - Nutcracker fractures of the tarsal navicular and cuboid usually result from dislocations of the midtarsal (Chopart's) joint. The classic pathomechanism consists of forced adduction or abduction (medial or lateral stress) mostly in combination with axial force. The resulting injuries are frequently misjudged or underestimated with respect to their potential consequences for global foot function. Radiographic imaging includes plain radiographs in exact standard projections and computed tomographic scanning. Classification of Chopart fracture dislocations is based on the proposed pathomechanics and the direction of the dislocating force. The goals of open reduction and stable internal fixation of Chopart fracture-dislocations are realignment of the medial and lateral columns of the foot, restoration of joint congruity and temporal transfixation in case of ligamentous instability to ensure proper ligament healing. PMID- 15354755 TI - [Tarsometatarsal injuries--an often neglected entity]. AB - Tarsometatarsal dislocations or fracture dislocations represent infrequent, but severe injuries which endanger the structural and mechanical integrity of the midfoot if the diagnosis is missed initially. Delayed diagnosis may result in painful and disabling arthritis and the need for salvage reconstructive surgery. As such, the rationale of treatment should follow the principles of reconstruction of weight-bearing joint injuries. The degree of instability and dislocation will guide the decision for surgical intervention and anatomic reconstruction. As the clinical features of Lisfranc injuries are rarely conclusive an adequate radiographic examination of the foot employing three standard projections (dorso-plantar, lateral and 45 degrees oblique) usually supplemented by CT scans and/or MRI is decisive for a correct analysis of the injury components and an optimum selection of treatment options. Anatomic reduction and alignment are prerequisites for a good functional outcome. The reduction of the second metatarsal ray is the keystone and the first step of surgical reconstruction followed by the other structures involved. Since adequate stability is needed until definite healing has taken place the temporary transfixation of the corresponding tarsometatarsal joints employing small fragment positioning screws has substantial advantages compared with the traditional temporary K-wire arthrodesis. PMID- 15354756 TI - [Fractures of the midfoot]. AB - The immediate reduction of luxations and severe dislocations is necessary to protect the soft tissue. The soft tissue lesions determine the timing and partially the options of the operative treatment too. The conventional standard X rays are sufficient to classify most of the fractures. Conservative treatment or minimal-invasive intramedullary techniques of fixation are sufficient in the treatment in most of the fractures. Screw- and plate-osteosynthesis are a good choice only in intra-articular fractures or sometimes in fractures of the first or fifth metatarsals. The clinical outcome is almost good and the rate of complications is low. PMID- 15354757 TI - [Does acyclovir prevent Bell's palsy in post surgical therapy for otosclerosis?]. PMID- 15354758 TI - [Multidetector computed tomography: a new change for diagnostic protocols]. PMID- 15354759 TI - [Therapeutic effect of the association between pulmonary ventilation and aerosol- inhalation with sulphureous mineral water in the chronic bronchopneumopathies]. AB - AIM: Our research evaluates the efficacy of the inalation therapy by mineral sulphureous water in patients suffering from cronic bronchopneumopathy. MATERIALS AND METHODS: From August to October 2002, two groups of patients, randomly selected, suffering from cronic bronchopneumonopathy, were studied at spa center "Pompeo" in Ferentino (FR). Every patient was treated daily during a cycle of twelve days. The first group was treated by pneumonic mechanical ventilation associated to sulphureous water aerosolinalation, the second by mechanical pulmonary ventilation associated to sulphureous water aerosol-inalation medicated with flunisolide. Every patient carried out spirometry, before and after treatment, and the measurement of FEV1 was considered a good parameter to evaluate the respiratory function. RESULTS: All patients reported an improvement in symptoms as dyspnea and cough. Results obtained show a significant functional improvement of respiratory function in both groups, evaluated by FEV1. Equal efficacy treating the patients by medicated and not medicated treatments stress the therapeutic role of mineral sulphureous waters in chronic bronchopneumopathies. Our research points out the usefullness of pneumonic ventilation associated with sulphureous water aerosol-inalation in patients suffering from chronic bronchopneumopathies. PMID- 15354760 TI - [Mud and baththerapy in the acne cure]. AB - The skin, it is well know, is an organ of frontier and it is natural seat to natch, or collition, between exogenous stimoli and endogenous responses. Termal water and spud mud revelead their better potency on the skin and for that their tird close attention in dermatology. Our primary pourpose was to evalue eventuals modifications induced by thermal water and mud on healty skin and to know how their alter in significative mode functionals parameters, an alteration of which characterising pathogenetics events in development of acne. They are: follicular keratinisation, sebum synthesis by sebaceum glands, skin inflammation, loss of gate function of epidermidis. Results are showed. PMID- 15354761 TI - [Tardive dyskinesia: review of the literature]. AB - PURPOSE: Tardive Dyskinesia (TD) is an adverse effect of several psychotropic drugs for long-term therapy for schizophrenia and other psychoses. TD is considered an important issue for public health for its invalidating characteristics. An extensive review of the literature on studies concerning efficacious treatments for TD has been carried out. MATERIALS AND METHODS: A computer-generated search of the biomedical literature up to and including December 2002 was undertaken to identify studies. RESULTS: Some evidence supports the switch from typical neuroleptics to atypical antipsychotics in patients with TD (particularly clozapine and olanzapine) and supports the use of atypical antipsychotics as first choice treatments. Other than the new antipsychotics, the gabaergic compounds, the new anticolinergics and some antioxidants as melatonin seem to be clinically useful. However, their tolerability and efficacy have to be confirmed by randomized clinical trials. CONCLUSIONS: The recent introduction of atypical antipsychotics seems to reduce the risk of TD in patients with long-term and maintenance therapies. Available literature data show that TD is less frequent with the new antipsychotics but still remains a well-known complication. The future research sould be directed to confirm the efficacy of these new drugs and to offer other treatment opportunities for patients that cannot change typical with atypical antipsychotics or for patients where the onset of the TD appears during therapy with atypical compounds. PMID- 15354762 TI - [Spasticity in multiple sclerosis]. AB - Already in 1860, the great neurologist Charcot described the symptom spasticity in patients affected by "sclerose en plaque". Spasticity is one of the most common symptoms of multiple sclerosis MS). The consequences of spasticity are very disadvantages because it hinders the functional mobility and overburden disability. Moreover, in the later stages of MS spasticity may be complicated by seating problems, pressure sores, fibrous contractures and poor perineal hygiene. In this article, the therapeutical management of MS spasticity, in all its components (pharmacological, rehabilitative, surgical) is reviewed. PMID- 15354763 TI - [Drug/food interactions: an actual therapeutic outcome]. AB - The biological effects of specific interactions occurring between drugs and foods may greatly vary, determining severe side effects in some people. A lot of factors may influence such interactions, i.e. age, sex, dietary composition, and so. In addition, further interaction may be determined by cigarette smoking, alcohol abuse and coffee consumption. Older people, as well as children, chronic ill and malnourished people seems to be most commonly affected by side effects due to food/drugs interaction. The aim of this review is to give a synopsis of the drug/food interactions most commonly occurring in clinical practice. PMID- 15354764 TI - [A case of double ectopic pregnancy dealed with a laparoscopic management]. AB - Uterine horn pregnancy is a very rare condition and is associate with a high rate of maternal morbidity and mortality. The standard treatment is laparotomy. CASE: A 40-year-old woman, previosly undegone laparotomy surgery for extrauterine pregnancy, was admitted to hospital "Casa di Cura Malzoni" because ectopic pregnancy was diagnosed. The patient was treated with laparoscopic surgery. PMID- 15354765 TI - [Strategy for safe nutrition in swallowing disorders]. AB - Swallowing deficits may affect the ability of patients to feed properly and sufficiently, therefore inducing malnutrition. In order to provide effective and durable treatment, a team of experts should evaluate the swallowing function of each individual patient. Treatment protocols must be carefully analyzed, and adapted to progression of the disease. We refer paradigmatically to patients with Huntington disease, although indications may be conveniently extended to other patients with impaired transit of food. PMID- 15354766 TI - [Claude Bernard: a personal scientific fame that he probably would have not appreciated]. AB - Claude Bernard has been a great medical physiologist of the ninenteenth century, who was able to convince biologists and physicians that the "experimental method" can and must be applied to the phenomena of living matter, as it is applied in physics and chemistry. With the concept of the "universality of the experimental method", the Modern Experimental Medicine begun to exist. However, C. Bernard deserves of further honor, having identified and promulgated a law of the living matter, i.e., the law concerning the "constancy of the interior biological environment". Such a law was later consacrated in 1929 by G.W. Cannon with the term "homeostasis". The homeostatic conception of biological functions had a great epistemological impact in the history of biology and medicine. As a matter of fact, all the modern biologists and physicians are today educated at the light of the homeostatic systems. Accordingly, C. Bernard is celebrated in his fame as the tutor of scientific medicine. Reading, however, what C. Bernard has written on the personalism and the eponimy of scientific ideas, one is prone to think that he would have not appreciated the dogmatic exaltation, the cult of his ideas on the biological constancy. He was aware that "... it is a fundamental rule in science to modify and change the ideas going on". And again "... But, since these ideas are not immutable verities, one must be ready to abandon and modify them when they cease to represent the reality". And again "The systems and doctrines are personal and pretend to be immortal and to preserve their personality. The experimental method, however, is impersonal; it zeroes the individuality and sacrifices the particular ideas of the a single person in favor of a general verity, which is established by means of the experimental criterion". And again "The great persons have never been promoters of absolute ed eternal verities in experimental sciences". And again "Any person who made the science to proceed, never pretended to define the limits; this person is destined to be surpassed and left apart by the progress of the future generations". And again "One of the most important obstacle that is encountered by humans along the course of knowledge is represented by the tendency of human acquisitions to be transformed into systems". With the discovery of chaos and biological systems in living matter, the law of homeostasis must be now revised. PMID- 15354767 TI - [Study on loss of heterozygosity at microsatellite locus in larynx and pharynx squamous cell carcinoma and neck lymph nodes]. AB - OBJECTIVE: To access prognosis and explore neck lymph nodes micrometastasis in larynx and pharynx squamous cell carcinoma (LPSCC) with loss of heterozygosity (LOH). METHOD: By polymerase chain reaction (PCR), 18 cases of LPSCC and 72 resected neck lymph nodes were investigated with LOH at D9S171 locus. RESULT: The higher level of LOH (40.28%) was found in neck lymph nodes compared with the positive level of neck lymph nodes detected by pathological examination (8.33%) (P < 0.01); LOH was related with tumor differentiation in patients with LPSCC (P < 0.05). The LOH rate of primary tumor was closely correlated with the clinical stages (P < 0.01). CONCLUSION: Our results suggested that analysis of LOH at D9S171 locus is one of sensitive means of evaluating neck lymph node micrometastasis, and which may be one of prognostic factors in patients with LPSCC. PMID- 15354768 TI - [Expression of matrix metalloproteinase-2,9 in hypopharyngeal carcinoma and its relationship with lymphatic metastasis]. AB - OBJECTIVE: To study the relationship between matrix metalloproteinase-2,9 (MMP 2,9) and the biology behavior, lymphatic metastasis of hypopharyngeal carcinoma by investigating the expression of MMP-2,9. METHOD: Two-step immunohistochemical method was used to examine the expression of monoclona antibody in hypopharyngeal carcinoma tissues. RESULT: It was found that the positive rate of MMP-2 in 32 hypopharyngeal carcinoma tissues was 53.12% (17/32),and the positive rate of MMP 9 was 59.38% (19/32). The difference of the expression in tumor tissue and normal mucosa beside the tumor was significant. The positive rate in those carcinoma with cervical lymph node metastasis was much higher than that in cases without cervical lymph node metastasis. And there was no significant difference found in the expression of MMP-2 and MMP-9. CONCLUSION: MMP-2 and MMP-9 may relate with the lymph node metastasis in hypopharyngeal carcinoma. PMID- 15354769 TI - [The gene expression of Th1/Th2 cytokines and its role in hypopharyngeal carcinoma]. AB - OBJECTIVE: To observe the mRNA expression of Th1/Th2 cytokines in hypopharyngeal neoplasms and to explore the pathogenesis of it. METHOD: The gene expression of Th1/Th2 cytokines in patients with hypopharyngeal neoplasms was detected by RT PCR. RESULT: The cytokines expression of Th2 was significantly stronger than Th1 in tissue of hypopharyngeal carcinoma. CONCLUSION: The dominant expression of Th2 cytokines in hypopharyngeal neoplasms may inhibit the human immunity and enhance the growth and metastasis of hypopharyngeal carcinoma. PMID- 15354770 TI - [Surgical management of carcinoma of the hypopharynx and cervical esophagus]. AB - OBJECTIVE: To report our experience in reconstructing hypopharynx and cervical esophagus with four kinds of operations. METHOD: Twenty one patients underwent reconstruction of the hypopharyngeal and cervical esophagus defect using contralateral hemilaryngeal mucosa flap, pectoralis major myocutaneous flap, forearm free flap and jejunal free flap. RESULT: All of cases were successful. Postoperative pharyngocutaneous fistulas occurred in two patients, which resolved spontaneously. All of patients can swallow ordinary food and had no benign strictures after operation. CONCLUSION: Every operation style has advantage and disadvantage. We had to select the operation of choice depending upon the location and size of the carcinoma, perioperative complication. PMID- 15354771 TI - [First stage speech reconstruction after total laryngectomy with the improved trachea-pharynx anastomosis]. AB - OBJECTIVE: To investigate the improved methods of first stage speech reconstruction after total laryngectomy. METHOD: We improved the method of the first stage speech reconstruction after total laryngectomy with a low level tracheotomy, reconstruction of the glottis with the ahead mucous membrane of esophagus, making stoma at the incision of trachea, lifting and shrinking the cervical trachea and sewing up to the root of tongue. This novel method was adoptioned on 35 patients, the curative effect had been observed. RESULT: Among the 35 patients, 33 patients (94.29%) regained the phonation function to a different extent. Only 2 patients (5.71%) could not speak. Most patients got an articulate and fluent voice. All of the patients could take food by mouth without severe miss deglutition. CONCLUSION: There are many excellences in our operation style. The success rate of the speech is higher, the quality of the speech is better and the rate of the miss deglutition is lower than the classical operation style of the speech reconstruction. It has no use for a factitious speech device and deserves to spread. PMID- 15354772 TI - [Application of platysma myocutaneous flaps reconstructing esophagus in stenosis of cervical esophagus]. AB - OBJECTIVE: To observe the application of platysma myocutaneous flaps (PMF) reconstructing esophagus in stenosis of cervical esophagus. METHOD: Retrospective analysis was carried out in 33 patients with stenosis of the cervical esophagus reconstructed by PMFs in our department from April 1990 to January 2003. The patients were followed-up for 6 to 156 months with esophagography and esophagoscopy. The pathological changes of the flaps were examined with biopsy in 4 cases. RESULT: There was no operative death. All the flaps survived. Three patients developed fistula of cervical anastomosis after operation and healed with conservative method. Stricture of cervical anastomosis occurred in 2 cases, one patient received Dilatation and one treated by reoperation. The transferred flaps had no ulcer and cancer in long time follow-up. The squamous epithelial lawyer of the skin is thinner, but keep keratinization. All the patients were able to take solid foods. CONCLUSION: The advantages of repairing stricture of cervical esophagus with PMF is that the operative wound is small, the transference is easy and can be completed in one stage, the postoperative complication is low, the long-term result is excellent. The PMF is one of good option for cervical esophageal reconstruction. PMID- 15354773 TI - [The observation on the mechanism of speech forming by slit-like trachea esophagus puncture after total laryngectomy]. AB - OBJECTIVE: To observe the mechanism of the rehabilitated speech using slit-like fistula between trachea and esophagus after total laryngectomy. METHOD: Forty patients after total laryngectomy all come from the fourth central hospital of Tianjin. They all reconstructed voice using slit-like fistula between trachea and esophagus primary or secondary stage. We use teleradiography and laryngotroboscope to observe its mechanism of store gas stage, deliver gas stage and phonation stage. RESULT: While inspiratory, the pulmonary enlarged, and the upper esophagus was closed and circulus invension. After this, the infra pharyngeal cavity expanded, looks like a downward triangular taper. The upper end of the esophagus opened suddenly. Phonation begin, the superior esophagus was full of air as a ball. The air escape through the segment of trachea-esophagus. The meatus of esophagus opened in coronal position and turn over like lip. It is vibrated together with nearby mucus, mucosa to form voice. After that, thorax shrink, the meatus of esophagus turn in again, the voice behavior is over. CONCLUSION: The power organ of trachea-esophagus voice is lung. It is crucial for the trachea-esophagus fistula unobstructed and smooth to forming voice. When phonation, the mucosa, mucus of infra-pharyngeal and esophageal meatus take part in vibration. PMID- 15354774 TI - [The relationship between the surgical margin and local recurrence of laryngeal carcinoma]. AB - OBJECTIVE: To study the relationship between the surgical margin and local recurrence of laryngeal carcinoma. METHOD: The surgical margin, T staging, postoperative irradiation and the local recurrence of 160 cases of laryngeal carcinoma operated between 1991 and 1999 were retrospectively reviewed and analyzed. RESULT: (1) Of 160 cases of laryngeal carcinoma, local recurrences were occurred in 36 cases (22.5%). (2) Local recurrence was statistically significantly related to the T staging of the tumor. (3) By comparing the local recurrence of 3 groups of patients: surgical margin < or = 3 mm, between 4-5 mm and > 5 mm, the rate of local recurrence of the group with surgical margin less than 3 mm were significantly higher than that of the groups with surgical margin between 4-5 mm and > 5 mm. (4) Of 69 cases with surgical margin less than 3 mm, 32 cases were received the postoperative irradiation, while 37 cases did not undergo. Statistical analysis showed that the local recurrence of the group of patients received postoperative irradiation was significantly lower than that of patients who did not received postoperative irradiation. (5) By analyzing the rate of local recurrence of difference surgical margin in the patients with glottic and supraglottic carcinoma, we found that the 5 mm as a surgical margin was relatively safe for the glottic carcinoma, but it was not the case for the supraglottic carcinoma. CONCLUSION: 5 mm as a surgical margin is relatively safe for the glottic carcinoma, but it is not the case for the supraglottic carcinoma. Therefore, the surgical margin more than 5 mm is proposed for supraglottic carcinoma. For the patients with surgical margin less than 3 mm, postoperative irradiation can effectively reduce the rate of local recurrence, but this does not mean that we can reduce the standard surgical margin during the laryngectomy. PMID- 15354775 TI - [The causes and management of laryngostenosis in clinical stage I, II laryngeal cancer patients after decannulation]. AB - OBJECTIVE: To investigate the causes and management of laryngostenosis in clinical stage I, II laryngeal cancer patients after decannulation. METHOD: The causes of seventeen cases laryngostenosis after decannulation were analyzed and various operations were carried out according to the causes. Two cases were treated by laser through fibrolarnygoscope, granulation tissue in laryngeal cavity were resected by selfretcuining laryngoscope in two patients. The laryngeal cavity scar was removed and the epiglotic flap, bi-pedicled myoperichondral flap, platysma myocutaneous flap were utilized to reconstruct the laryngeal functions in ten patients. Three patients were treated by total laryngectomy. RESULT: Except for three cases of total laryngectomy, thirteen cases were decannulated successfully after the second operation, decannulation rate was 92.86%. CONCLUSION: The scar hyperplasia and granulation tissue are the major causes of secondary laryngostenosis after decannulation in clinical stage I, II laryngeal cancer patients. Most of them regained own respiration, swallow and voice function. PMID- 15354776 TI - [Prevention of complications during paralaryngeal space neoplasms surgery]. AB - OBJECTIVE: To explore the prevention of complications during the the paralaryngeal space (PPS) neoplasms surgery. METHOD: A retrospect study was carried out from 41 patients, suffering from paralaryngeal space neoplasms and recerving surgery from 1990 to 2002. The surgical procedures included oral approach in 6 cases, parotid approach in 6 cases, cervical approach in 25 cases, cervical approach with mandibular swing in 2 cases and lateral skull-base approach 2 in cases. Eight cases got the tracheodectomy during the operations. The tumor in 37 cases were resected throughly, except for 4 cases with partial resection. RESULT: The patients were followed up for 11 months to 6 years. The surgical complications consists of surgical cavity infections in 2 cases, a massive bleeding in 1 case with huge nerve sheath tumor and permanent nerves damage in 7 cases, including Horner's syndrome (2 cases) and nerves damage (5 cases), such as VII, IX, X and XI. Six cases relapsed: 1 case with benign tumor through oral approach and 5 cases with malignant neoplasms. CONCLUSION: Familiarity with PPS anatomy, preoperational evaluation in detail, proper surgery approach and perfect treatment during the operation are vital important to avoid the complications in PPS neoplasms. PMID- 15354777 TI - [DNA quantitative analysis of head and neck tumor]. AB - OBJECTIVE: This research discussed the diagnostic value of cellular quantitative analysis in head and neck tumor. METHOD: We analysis quantitative parameters of 23 malignant tumors and 20 benign diseases with automated DNA image cytometry, they are the DNA Index (DI), S-phase Fraction (SPF), Proliferation Index (PI) and 5c Exceeding (5c Ex). RESULT: The masculine rate of cellular quantitative analysis and conventional method are 76.92% and 50.00%. The value of SPF, PI and 5c Ex are 14.05%, 19.59% and 1.59% in malignant tumor, respectively. The parameters of SPF and PI showed distinct difference in the area of malignant tumor than the 1 cm sideward area of tumor and the edge of tumor. CONCLUSION: Automatic DNA image cytometry has higher masculine rate of malignant tumor than conventional method in early detection of tumor. The cellular quantitative parameters, such as SPF, PI and 5c Ex, are able to guide clinical work, specially in diagnosing of head and neck tumor, estimating biological character of malignant tumor and evaluating the area of excision. PMID- 15354778 TI - [Permanent interstitial implantation of 125I seeds for 5 cases of advanced head and neck cancer]. AB - OBJECTIVE: To observe the effect of 125I seeds implants in advanced head and neck cancer. METHOD: Treatment fields were designed to encompass the tumor/edema volume with a 2-3 cm margin, based on assessment of diagnostic CT images, 125I seeds were inserted into there and permanent left there for brachytherapy in 5 patients with advanced head and neck cancer including recurrent oropharyngeal cancer, supraglottic cancer and occult cancer. RESULT: Local tumor were controlled evidently and the patients suffering were alleviated, but 1 case occurred skin necrosis and 1 case occurred skin necrosis combined with internal carotid rupture. CONCLUSION: Permanent 125I implants offer useful pallition for patients with advanced recurrent head and neck cancer but need prevention fatal complication. PMID- 15354779 TI - [Small partial laryngectomy for early-stage glottic carcinoma]. AB - OBJECTIVE: To explore the surgery way for T1, T2 glottic carcinoma. METHOD: Small partial laryngectomy were performed to 112 patients with T1, T2 (T1 N0 M0, T2 N0 M0) glottic carcinoma. The surgery method, effect and treatment advantage was summarized and compared with laser therapy and radiotherapy, the dynamic follow up of arytaenoidea cartilage movement, glottidis rimae conformation and voice change was analyzed. The preoperative and postoperative voice quality was compared with the software of Dr. speech system for windows. RESULT: All incisions were healed one stage in 6 or 7 days, the mean time in hospital is 9.76 days; all patients took food with mouth after 2 or 3 days, all trachea cannula was removed successfully during in hospital, the mean time with cannula is 7.32 days and the rate of removing cannula 100%. Arytaenoidea cartilago movement of 47 cases 1 week after surgery weakened and gradually improved, glottidis rimae conformation is close to be normal after 2 months. There is no significant difference of Shimmer and NNE between preoperative and postoperative 1 week (P > 0.05), but there is a significant difference between preoperative and postoperative 2 months, half a year, and also is between postoperative 1 week and 2 months, half a year (P < 0.01). All 76 cases survive during 3 years' follow-up, 35 of 36 cases in 5 years' follow-up survive (1 case died without definite cause), and there is 2 cases for recurrence. CONCLUSION: Small partial laryngectomy can provide large operative view, resect the tumor completely, make diet and speech recover in shorter time and improve the voice quality, so the surgery is available. PMID- 15354780 TI - [Malignant change of laryngeal papilloma in adults]. AB - OBJECTIVE: To explore the canceration of laryngeal papilloma in adults, especially its pathological figures, clinical characters and long-term therapeutic efficacy. METHOD: Among 17 cases of laryngeal papilloma with malignant changes, total laryngectomy and single functional neck dissection were performed for 4 cases (glottis type T3 N1 M0 1, Supraglottic type T4 N0 M0 1, Subglottis type T4 N1 M0 2), partial laryngectomy were performed for other 13 cases. All patients received appropriate radiotherapy whose dose is 45-55 Gy. RESULT: Tracheal cannulas of all patients (17 cases) were pull out one year after operation. The rate of extubation is 100%. All patient were follow-up more than 5 years. Among 4 cases of total laryngectomy, one patient died from tumor recurrence 2 years after operation, one patient died from distant metastasis 3 years and 4 months after operation, two patients still alive without tumor 8 years after operation and voicing by using esophagus. Among 13 cases of partial laryngectomy, 12 patients still alive and 1 patient loss follow-up 3 years after operation; 11 patients still alive and 1 patient died from distant metastasis 5 years after operation. CONCLUSION: It is important that repeatedly biopsies to avoid fail to report the malignant change of laryngeal papilloma. The key to cure is complete resection of tumor. PMID- 15354781 TI - [18F-FDG PET in the diagnosis of head and neck tumor]. AB - OBJECTIVE: To analyze the value of 18F-FDG PET in the diagnosis of patients suspected of having head and neck malignant tumor. METHOD: Seventy-five cases patients suspected of having head and neck cancer or recurrence were analyzed retrospectively. 18F-FDG PET was performed in all cases. The results of some cases were compared with CT and MRI. RESULT: 18F-FDG PET yielded a sensitivity 91.30% and specificity 76.92% and accuracy 86.11% in the diagnosis of nasopharyngeal carcinoma; sensitivity 100.00% and specificity 83.33% and accuracy 94.44% in esophageal carcinoma; sensitivity and specificity and accuracy 100% in thyroid carcinoma, laryngeal carcinoma and hypopharyngeal carcinoma. CONCLUSION: Preliminary clinical application of 18F-FDG PET has demonstrated its significant advantages in evaluating head and neck malignant diseases. PMID- 15354782 TI - [Evaluate the effect of Nd:YAG laser in treatment of obstructive sleep apnea hypopnea syndrome]. AB - OBJECTIVE: To observe the effect of laser in treatment of obstructive sleep apnea hypopnea syndrome (OSAHS). METHOD: Eighty-nine cases of OSAHS were treated by Laser assisted uvulopalptoplasty (LAUP). Among the cases, the soft palate and uvula were partly resected. The cureiform incise was made into the soft palate along the each side of uvula. RESULT: The follow-up period was six months, 76 cases received the detection of polysomnography. Eight cases were cured; 36 cases had notable effects; 25 cases had effects; 7 cases had no effects. The total ratio of validity was 90.8%. CONCLUSION: It has clinical value to use LAUP in treatment of OSAHS, and its advantage include hemostasia, solidfication, penetrability, clear visual threshold, lower complication and shortening the surgery time. PMID- 15354783 TI - [The immediate local skin flaps transference for the treatment of acute auricle trauma defect]. AB - OBJECTIVE: To investigate the advantage and postoperation notice of immediate local skin flaps transference for the treatment of acute auricle trauma defect. METHOD: A retrospective study was carried out to judge the immediate local skin flaps transference for sixty-three patients who underwent ear injury. RESULT: All the flaps survived postoperation. After a follow-up observation for 12-18 months, the shapes of all patients' ears were satisfying, and there were no osteochondritis. CONCLUSION: Immediate local skin flaps transference for the treatment of acute auricle trauma defect could recover the shape of ears in short time. The colour of flaps were similar as the ears, so the local skin flaps were ideal material for the ear reconstruction. If the wound were contaminated, antibiotics must be used, in addition to the Dexamethasone intravenous for seven to ten days which may inhibit the osteochondrites to some extent. PMID- 15354784 TI - [Cholesterol granuloma media: 8 cases report]. AB - OBJECTIVE: To investigate the etiology, pathogenesis diagnosis and therapy of cholesterol granuloma media. METHOD: Eight cases (9 ears) of middle ear cholesterol granuloma were retrospectively reviewed. Nine ears all have a speciality of conductive deafness with a light to middle level, blue eardrum and tympanogram all showing B type (except one case of eardrum perforation in the tension part). Seven cases (8 ears) were cured by tympanic cavity exploration opening epitympanum, tympanic antrum, mastoid cells and thoroughly eliminating granulation tissue. Grommet insertion and modified radical mastoidectomy were performed on 5 cases and 2 cases, respectively. The other one case was only carried out with eardrum puncture. RESULT: Seven cases (8 ears) after operation were followed up for six monthes to six years. Six cases (7 ears) were cured out without recurring; one case (1 ear) recurred. One case was not followed up. CONCLUSION: Otoblennorrhea of blood without obvious reason and blue eardrum should be coupled with CT to enhance the diagnosis rate before operation. CG should be treated by operation to clean disease location, air and drainage tympanicus cavity and mastoid. PMID- 15354785 TI - [Compatibility research of self-designed scaffold biomaterials of nature extracellular matrix]. AB - OBJECTIVE: Scaffold biomaterials for cell culture of tissue-engineering demand satisfactory biocompatibility. This experiment aimed to evaluate biocompatibility of self-designed scaffold biomaterial of NECM. METHOD: The biocompatibility of self-designed NECM was evaluated both in vivo and in vitro, including cytotoxic test, acute and subacute systemic toxicity test, hemolysis test, pyrogenic test, lymphocyte transformation test and long-term subcutaneous implantation test of NECM. RESULT: Self-designed NECM scaffold biomaterial for cell culture has no cytotoxicity, no acute and subacute systemic toxicity, and it did not cause inflammatory reaction. CONCLUSION: Self-designed NECM has good biocompatibility and may become an ideal biological scaffold material for cell culture of tissue engineering. PMID- 15354786 TI - [Study on the expression of eIF families after elemene treatment]. AB - OBJECTIVE: To investigate the activity of Caspase-3 and the expression of eukaryotic initiation factor families, bFGF and VEGF after elemene on laryngeal carcinoma HEp-2 cells. METHOD: The HEp-2 cells after elemene treatment were analyzed utilizing Westernblot and reverse transcriptase polymerase chain reaction (RT-PCR). The activity of Caspase-3 was assessed by colorimetric assay. RESULT: The activity of Caspase-3 was enhanced after elemene treatment. The protein expression of eIF4E, eIF4G, bFGF and VEGF were significantly inhibited by elemene; and the mRNA expression of bFGF and VEGF were inhibited either. CONCLUSION: Elemene can effectively inhibit the growth of HEp-2 cells and result in the alteration of activity of Caspase-3. There were significant correlations between the decreased expression of protein eIF4E, eIF4G, bFGF and VEGF. The mechanism of eIF4E and eIF4G decrease the expression of bFGF and VEGF is post transcriptional. PMID- 15354787 TI - Physicochemical traits of Dekoko (Pisum sativum var. abyssinicum) seeds. AB - Dekoko (Pisum sativum var. abyssinicum) has high appreciation for its taste and obtains a premium price in local markets compared to Ater (Pisum sativum var. sativum). However, data on the physicochemical traits of Dekoko seeds were lacking. This paper reports on the physicochemical features of Dekoko and compares the results with that of Ater. Seed weight and seed volume were 36 and 30%, respectively, higher in Ater, while water absorption, percent seed swelling, and percent husk were higher in Dekoko. Cooking time was shorter for Dekoko than Ater seeds. Decortication reduced cooking time on average by 39 and 45 min in Dekoko and Ater, respectively. Cotyledon flour of Dekoko contained 251 g crude protein, 19 g fat, 31.7 g total sugars, 370 g starch, and 130 g neutral detergent fiber per kilogram DM. These traits were significantly higher in Dekoko than in Ater, except for starch, which was higher in the latter. Arginine, asparagine, and glutamine occurred in larger proportions, and collectively contributed about 39% to the total amino acids in both varieties. Lysine contributed about 7%, while sulfur containing amino acids constituted about 3.0 and 2.3% of the total amino acids in Dekoko and Ater, respectively. Ca and Mg were higher in Ater, while P was higher in Dekoko. Based on the observations it was concluded that Dekoko is a suitable supplementary protein source for a cereal-based diet. PMID- 15354788 TI - Association between seed coat polyphenolics (tannins) and disease resistance in common bean. AB - Common beans (Phaseolus vulgaris L) contain a number of antinutritional factors such as condensed tannins. Reducing tannin concentration might contribute to improving the nutritional quality of common bean. But polyphenolics are involved in resistance to diseases and pests, and reducing tannin concentration may have a negative effect on plant resistance. Furthermore, the effects of tannin on disease resistance in different gene pools or in different seed colors are not defined. To investigate these effects, 790 accessions from a common bean core collection were investigated. Data were subjected to independent sample t-tests, and the calculation of correlation coefficients. The mean coat extracts of black and red bean classes were highest (with 0.129 g/g and 0.124 g/g of seed coat, respectively). Among the gene pools, the coat extract was greater in the Middle American gene pool (0.129 g/g) than in the Andean gene pool (0.108 g/g). Coat extract in the Andean gene pool was positively correlated with susceptibility to Middle American isolates of anthracnose and to common bacterial blight, but negatively correlated with susceptibility to Andean isolates of angular leaf spot and to empoasca. Only empoasca damage showed negative correlation with coat extract in the Middle American gene pool. However within gene pools, the coat extracts of different seed classes varied in correlations with reactions to disease and pest infestations. Significant correlations were particularly associated with the black seed class in both gene pools. The relationships between coat extract and disease reactions are complex. A better understanding will help breeders to select germplasm with improved nutritional quality without adversely affecting disease resistance. PMID- 15354789 TI - Nutritional evaluation and shelf life studies of papads prepared from wheat legume composite flours. AB - The present study was carried out to develop papads from wheat and legume blends and to analyze them for organoleptic acceptability, nutritional quality, and keeping quality. Papad is a dehydrated product prepared from dhals or rice. A firm but pliable dough is made from the flours of dhals or rice with addition of suitable seasoning. It is shaped into balls, rolled out thin, dried and toasted over open fire so as to give a light, crisp product. Mung flour papads were kept as control and wheat flour, chickpea flour, and pea flour were used to supplement mung flour papads. Three different proportions (10, 20, 30%) of each flour were used to supplement Mung flour. Papads with wheat flour (10%), chickpea flour (20%), and pea flour (10%) level of supplementation were found to be most acceptable and these papads were subjected to nutritional evaluation. Protein content significantly increased on supplementation with legume flours at all levels. Fat content was significantly higher in chickpea flour supplemented papads. Ash content varied significantly from 10.17 to 10.78% in papads. Total carbohydrates decreased significantly on supplementation with chickpea flour. Copper content increased significantly on supplementation. A significant decrease in phytic acid and trypsin inhibitor of supplemented papads was found. In vitro protein digestibility significantly increased on supplementation but a significant decrease in in vitro starch digestibility was found in supplemented papads. Storage studies showed that chickpea flour and pea flour supplemented papads can be stored safely for 60 days and wheat flour supplemented papads for 30 days both at room and refrigeration temperatures. PMID- 15354790 TI - Biochemistry of fruit ripening of guava (Psidium guajava L.): compositional and enzymatic changes. AB - Changes in chemical composition and the activities of hydrolytic enzymes during four different stages of maturity, viz. mature green (MG), color turning (CT), ripe (R), and overripe (OR), have been studied in guava fruits cv. Banarsi Surkha. Chlorophyll content decreased while carotenoid content increased during ripening. Starch content decreased with concomitant increase in alcohol-soluble sugars. Cellulose, hemicellulose, and lignin also decreased up to ripe stage, while pectin continued to decrease up to OR stage. PG (polygalacturonase) and cellulase exhibited progressive increase in activity throughout ripening, whereas pectin methyl esterase (PME) activity increased up to CT stage and decreased at R stage. The activities of alpha-amylase and beta-amylase decreased significantly with ripening. The most notable metabolic changes occurred between MG and CT stage, implying that for improved postharvest handling, guava fruits may be harvested at CT stage. PMID- 15354791 TI - Expression of ripening-related genes in prickly pear (Opuntia sp.) fruits. AB - To throw light on the expression of ripening-related genes in prickly pear (Opuntia sp.) fruits and on the possible role of the gaseous hormone ethylene in nonclimacteric fruit ripening, cDNA fragments that showed high homologies with 1 aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase cDNAs from other plants were cloned and partially characterized. Thus, the corresponding genes were accordingly named opaccs-1 and opacco-1, after Opuntia ACC synthase-1 and Opuntia ACC oxidase-1, respectively. Southern analysis suggests the presence of at least one copy of both genes, as well as other related homologous sequences in the Opuntia genome. Northern analysis of the opaccs-1 gene shows an enhanced expression in ripening fruit tissues, whereas opacco-1 expression is highly induced in ripe tissues with respect to the green fruits and mature cladodes. These results are in agreement with an active metabolic role of ethylene during nonclimacteric prickly pear fruit ripening. This is the first report on the analysis at the molecular level of ripening-related genes of the Opuntia genus. PMID- 15354792 TI - Sleep deprivation and medicine. PMID- 15354793 TI - A lexicographer's weary pilgrimage. PMID- 15354794 TI - Obesity as a risk factor for osteoarthritis: mechanical versus metabolic. AB - Obesity is a risk factor for the development and progression of radiograhic OA and probably works through both mechanical as well as metabolic pathways. We are just beginning to unravel these associations. By better understanding these associations, improved biomarkers for development and progression of OA and new novel therapies for the prevention and treatment of OA may be developed. The Osteoarthritis Initiative will help us understand these associations and allow us to evaluate new biomarkers for symptomatic incident and progression of established knee OA as well as the relationship of obesity and generalized OA. PMID- 15354795 TI - Pain in osteoarthritis. PMID- 15354796 TI - Genetic aspects of osteoarthritis. PMID- 15354797 TI - Current thinking on viscosupplementation in osteoarthritis. PMID- 15354798 TI - Images in medicine. An arterial dissection. PMID- 15354799 TI - Six Rhode Island hospitals collaborate to prevent surgical site infections. PMID- 15354800 TI - Hospitalizations by race and ethnicity, Rhode Island, 1990-2003. PMID- 15354801 TI - Miriam Hospital morbidity and mortality conference, a case of a painful knee. PMID- 15354802 TI - A physician's lexicon: the arcana, the obscure, and the esoteric. PMID- 15354803 TI - The push for more inspections of sedation practices. PMID- 15354804 TI - Clinical audit: introduction: getting started. AB - Clinical audit can benefit your practice and your patients. It is one of the pillars of clinical governance that will ensure that SAAD members deliver the best quality dental care. If you have experience of clinical audit that you would like to share with members, we would like to hear about it, so other members can learn from your experience. PMID- 15354805 TI - Medical device alerts. PMID- 15354806 TI - A useful technique for cannulation using a venflon cannula. PMID- 15354808 TI - Conscious sedation in the provision of dental care: new guidelines. PMID- 15354807 TI - Changes in drug labelling. Letter to all practitioners. PMID- 15354809 TI - Intravenous sedation in children. PMID- 15354810 TI - A model for consolidation of clinical microbiology laboratory services within a multihospital health-care system. AB - To determine the cost-effectiveness of consolidating clinical microbiology services in a three-hospital health-care network while maintaining high-quality laboratory services, a retrospective review of the total costs of maintaining separate clinical microbiology laboratories within our health-care system was compared to the cost of providing these services after consolidation. Turnaround times before and after consolidation were compared to assess efficiency of the consolidated services. Input of clinicians was also solicited to ensure that quality of services and customer satisfaction remained high. The results of the consolidation project show that the net fiscal saving because of consolidation of clinical microbiology services within our health-care system will be approximately 100,000 dollars per fiscal year. This value includes increased courier charges as well as personnel savings. Although fiscal savings are an integral part of any laboratory consolidation plan, the financial considerations must be balanced by quality of service. The response to consolidation from clinicians was decidedly mixed before implementation of the plan because of fear of increased turnaround times and limited access to laboratory information. The consolidation process, however, was smooth with few physician complaints. The consolidation of our clinical microbiology services illustrates that significant financial savings can be achieved without compromise of efficiency or quality of service. PMID- 15354811 TI - An analysis of reference laboratory (send out) testing: an 8-year experience in a large academic medical center. AB - BACKGROUND: Utilization of outside reference laboratories for selected laboratory testing is common in the United States. However, relatively little data exist in the literature describing the scope and impact of these services. In this study, we reviewed use of reference laboratory testing at the Massachusetts General Hospital, a large urban academic medical center in Boston, Massachusetts. METHODS: A retrospective review of hospital and laboratory administrative records over an 8-year period from fiscal years (FY) 1995-2002. RESULTS: Over the 8 years studied, reference laboratory expenses increased 4.2-fold and totaled 12.4% of the total laboratory budget in FY 2002. Total reference laboratory test volume increased 4-fold to 68,328 tests in FY 2002 but represented only 1.06% of the total test volume in the hospital. The menu of reference laboratory tests comprised 946 tests (65.7% of the hospital test menu) compared to 494 (34.3%) of tests performed in house. The average unit cost of reference laboratory tests was essentially unchanged but was approximately 13 times greater than the average unit cost in the hospital laboratory. Much of the growth in reference laboratory cost can be attributed to the addition of new molecular, genetic, and microbiological assays. Four of the top 10 tests with the highest total cost in 2002 were molecular diagnostic tests that were recently added to the test menu. CONCLUSION: Reference laboratory testing comprises a major component of hospital clinical laboratory services. Although send out tests represent a small percentage of the total test volume, these services account for the majority of the hospital laboratory test menu and a disproportionate percentage of laboratory costs. PMID- 15354812 TI - Branding an anatomic pathology practice to build revenue. AB - Innovative Pathology Services (IPS) is an Associate Practice of Pathology Service Associates (PSA). PSA is an organization known as the "Business Solution for Pathology." IPS provides pathology services to nine hospitals, including two large tertiary-care medical centers, a progressive and renowned children's hospital, a cancer survival center, five surgery centers, and numerous physician's offices and clinics throughout east Tennessee. We accept specimen referrals from other pathology practices and providers from across the country. The center of operations is in Knoxville, a mid-sized metropolitan district. Until January 1, 2003, we were known as Knoxville Pathology Group (KPG). We renamed our practice because KPG did not reflect our service area, was limiting by perception, barely distinguished us from other groups, and did not describe our culture and philosophy. IPS is a new name for a well-established pathology group with a solid foundation and a long history of providing services at the point-of-care. As such, we offer all services that we offered through our foundation practice, and, in addition, these services were enhanced and new services were added. Our entire "team" and, in particular, the pathologists, were involved in the successful "branding" of IPS. Whether you are an independent anatomic pathology or clinical laboratory or you are hospital based, you may benefit from our experiences detailed in this article. PMID- 15354813 TI - Designing and implementing a balanced scorecard: lessons learned in nonprofit implementation. AB - The balanced scorecard has been referred to as the management innovation of the century, and extensive articles have been written using case studies of organizations that use this performance measurement system. This article addresses the key issues of design and implementation with a step-by-step guide to how to design a balanced scorecard and lessons to avoid implementation problems in government and nonprofit settings. PMID- 15354814 TI - Opening laboratory communication. PMID- 15354815 TI - Using blood from autopsy cases for research. PMID- 15354816 TI - Common cents 19: The capital asset pricing model. PMID- 15354817 TI - Finding something you can live with: choosing laboratory casework. PMID- 15354818 TI - POL management. PMID- 15354819 TI - Neonatal administration of ACTH4-10 and its analog Semax to young laboratory mice modulates the number of catecholaminergic neurons in adult animal diencephalon. PMID- 15354820 TI - Neonatal administration of buspiron causes changes in intermale aggression of adult mice. PMID- 15354821 TI - A high-resolution area in the retinal ganglion cell layer of the Steller's sea lion (Eumetopias jubatus): a topographic study. PMID- 15354822 TI - The possibility of correction of the lipid profile of blood by superweak electromagnetic fields. PMID- 15354823 TI - The time-frequency structure of the spike-wave discharges in genetic absence epilepsy. PMID- 15354824 TI - Goiter as a biogeochemical problem. PMID- 15354825 TI - Modification of the immunomodulating effect of hydrocortisone under the conditions of beta-adrenoreceptor blockage. PMID- 15354827 TI - On the biotic self-purification of aquatic ecosystems: elements of the theory. PMID- 15354826 TI - Characteristics of fraction composition of the paddlefish (Polyodon spathula) hemoglobin. PMID- 15354828 TI - The striata morph and its role in the ways of adaptation of the genus Rana in the modern biosphere. PMID- 15354830 TI - Analysis of the growth capacity of embryo axes during seed dormancy and dormancy release. PMID- 15354829 TI - Carnosine modulates oxygen consumption and heat production by wheat root cells. PMID- 15354831 TI - Nonlinearity of climate-driven changes in phenological dates in woody plants. PMID- 15354832 TI - Animal growth and time. PMID- 15354833 TI - A new form of symbiotic bacteria Wolbachia found in the endoplasmic reticulum of early embryos of Drosophila melanogaster. PMID- 15354834 TI - State of skeletal muscles in larvae of sturgeons of the Volga Basin in artificial reproduction. PMID- 15354835 TI - Changes in the symptoms of hypothalamic diabetes insipidus after treatment with homologous exogenous DNA. PMID- 15354836 TI - Subfossil oribatid mites as the bioindicators of profound environmental changes during the holocene. PMID- 15354837 TI - Changes in carbohydrate and polyphenolics pools during potato tuber maturation: cultivar-specific development of resistance to pathogens. PMID- 15354838 TI - The role of isotopes in the evolution of some biological systems. PMID- 15354839 TI - Ultrastructural changes in interstitial Cajal's cells and myocytes of the small intestine of suckling rabbits with experimental cholera. PMID- 15354840 TI - Mycoplasma adaptation to biogenic and abiogenic stressful factors; Acholeplasma laidlawii nannotransformation and minibodies. PMID- 15354841 TI - Clasmosomes in Bezredka's phenomenon. PMID- 15354842 TI - Estimation of the death rate of 3T3 NIH cell at different phases of the cell cycle in chronic hyperthermia within the physiological temperature range. PMID- 15354843 TI - Effect of serotonergic receptor activation on the binding of [3H]quinuclidinyl benzylate by membrane muscarinic cholinoreceptors of the rat cerebral cortex. PMID- 15354845 TI - Chip shot (apology and letter of retraction). PMID- 15354844 TI - The Kendall Oration--Veterinarians in society--some future directions. PMID- 15354846 TI - Australian veterinarians who work with horses: attitudes to work and career. AB - OBJECTIVE: To describe the attitudes to their work and career of those Australian veterinarians who work with horses. METHOD: Questionnaires were mailed to 866 veterinarians who had been identified as working with horses, and 87% were completed and returned. Data were entered onto Excel spreadsheets, and analysed using the SAS System for Windows. RESULTS: The main attractions of veterinary work with horses were the horses themselves and the equine industries, but working outdoors and with rewarding clients, and the satisfaction of successful outcomes were attractions for many. The list of disadvantages was longer, and included unreasonable and disagreeable clients as well as those who provided inadequate facilities, could not control their horses or did not care for them. The physical demands and risks of injury, the amount of time required, low rates of return and difficulties in collecting payment, were other major disadvantages. Some mentioned concerns about litigation, unethical behaviour, and recruiting and retaining veterinarians competent with horses. For many in mixed practice, the difficulties in affording modern equipment, and of developing and maintaining their own competence with horses, was a real concern. More than three-quarters of the respondents reported that their careers had lived up to expectations and that they would become veterinarians again; 70% of equine veterinarians would become an equine veterinarian again. Almost all (93%) of the respondents were either very glad, or 'generally glad though with some misgivings' that they had done the veterinary course. Older veterinarians reported suffering less stress, and being more content with their career, than younger colleagues. CONCLUSIONS: The advantages of doing veterinary work with horses outweigh the disadvantages for most veterinarians, especially those well advanced in their careers. PMID- 15354847 TI - Multilobular osteochondrosarcoma of the hard palate in a dog. AB - A 14-year-old castrated male Rhodesian Ridgeback was presented with a history of sneezing and epistaxis. Diagnostic procedures included physical examination, regional and thoracic radiography, computed tomography and histological examination of an incisional biopsy. A multilobular osteochondrosarcoma of the hard palate with pulmonary metastases was diagnosed. Surgical resection of the primary tumour was achieved with clean margins and the defect was repaired using bilateral mucosal transposition flaps from the lips. Wound dehiscence and oesophageal stricture were postoperative complications, but these resolved with treatment. A long-term survival time of 14 months resulted, with good quality of life and function during this time. PMID- 15354848 TI - Hypokalaemia as a cause of recumbency in an adult dairy cow. AB - A 3-year-old Holstein cow was presented for evaluation of recumbency. Physical examination and laboratory evaluations resulted in a diagnosis of hypokalaemia causing extreme skeletal muscle weakness. Treatment involved intravenous and oral potassium supplementation, antimicrobial and anti-inflammatory therapy, and management of recumbency using a flotation tank (the Aquacow Rise System). The cow recovered and returned to the milking herd. Multifactorial elements were identified as the cause of hypokalaemia including inappetance, treatments for ketosis and administration of dexamethasone. PMID- 15354849 TI - Mandibular bone cysts in two horses. PMID- 15354850 TI - Persistent penile frenulum in a nilgai (Boselaphus tragocamelus). PMID- 15354851 TI - Implementation in Australia of molecular diagnostic techniques for the rapid detection of foot and mouth disease virus. AB - OBJECTIVE: To evaluate and implement rapid molecular diagnostic techniques for the detection of foot and mouth disease virus (FMDV) suitable for use in Australia. DESIGN: Two PCR TaqMan assays targeted to the FMDV internal ribosome entry site or the 3D polymerase coding region for the rapid detection of FMDV were evaluated using non-infectious materials to determine the test most appropriate for implementation as part of Australia's national preparedness for the rapid detection and diagnosis of FMD outbreaks. RESULTS: Two published tests (PCR TaqMan assays targeted to the FMDV IRES region or the FMDV 3D polymerase coding region) were evaluated for their ability to detect FMDV genetic material in non-infectious FMDV ELISA antigen stocks held at Australian Animal Health Laboratory. Both tests were able to detect FMDV genetic material from strains O1 Manisa, O-3039, A22, A24, A Malaysia, C, Asia 1 and SAT 1, 2 and 3. With the exception of Asia 1, the TaqMan assay targeted to the FMD 3D polymerase coding region had Ct values equal to or lower than for the TaqMan assay targeted to the IRES region suggesting that this test may provide broader serotype detection and sensitivity. However, the TaqMan assay directed to the FMDV IRES is the only one to date to have undergone substantial evaluation using clinical samples collected during an outbreak. The greatest differences observed were for O-3039, SAT 1, and 3. CONCLUSION: Given the ease of setting up both tests, AAHL currently runs both tests on highly suspect FMD investigations to provide independent confirmation of the absence of FMDV because the tests are focused on two independent regions of the FMDV genome. These tests add substantially to Australia's preparedness for FMD diagnosis complementing the already well-established virus isolation and antigen capture ELISA tests for index case diagnosis of FMD in Australia. PMID- 15354852 TI - Field evaluation of tracer sheep for the detection of early natural infection with Mycobacterium avium subsp paratuberculosis. AB - OBJECTIVE: To determine whether tracer sheep could be used to detect S strain Mycobacterium avium subsp paratuberculosis on pasture, and to provide further insight into the early stages of infection. DESIGN: A field study on two farms in an endemic area for ovine Johne's disease in New South Wales. Procedure Lambs, weaners and adult ewes were introduced to pasture with varying amounts of M. a. paratuberculosis contamination and monitored using skin tests, gamma interferon assay, faecal culture and serial necropsy of small groups for up to 15 months after first exposure. RESULTS: Culture from tissues was the most sensitive method for detecting early infection in sheep after natural exposure to S strain M. a. paratuberculosis. The organism was detected in at least one introduced sheep from every exposed group, 6 to 12 months after first exposure. Histopathological lesions were detected in only 17% of culture-positive sheep, and only after at least 8 months of exposure. Similarly, antemortem diagnostic tests had low sensitivity during the early stages of naturally acquired infection. There was no evidence of any differences in infection rate between sheep first exposed as neonates, as weaners or as adults. A higher proportion of lambs born to ewes from an infected flock were infected than lambs suckling uninfected ewes introduced to the same infected environment, and infection was detected earlier in these 'resident' lambs. CONCLUSION: These findings indicate that groups of unexposed 'tracer' sheep, tested by culture of tissues at slaughter 6 to 12 months after first exposure, might be a useful way to assess pasture infectivity in control programs for ovine Johne's disease. PMID- 15354853 TI - Effect of season and temperature on mortality in amphibians due to chytridiomycosis. AB - OBJECTIVE: To investigate the distribution and incidence of chytridiomycosis in eastern Australian frogs and to examine the effects of temperature on this disease. DESIGN: A pathological survey and a transmission experiment were conducted. PROCEDURE: Diagnostic pathology examinations were performed on free living and captive, ill and dead amphibians collected opportunistically from eastern Australia between October 1993 and December 2000. We conducted a transmission experiment in the laboratory to investigate the effects of temperature: eight great barred frogs (Mixophyes fasciolatus) exposed to zoospores of Batrachochytrium dendrobatidis and six unexposed frogs were housed individually in each of three rooms held at 17 degrees C, 23 degrees C and 27 degrees C. RESULTS: Chytridiomycosis was the cause of death or morbidity for 133 (55.2%) of 241 free-living amphibians and for 66 (58.4%) of 113 captive amphibians. This disease occurred in 34 amphibian species, was widespread around the eastern seaboard of Australia and affected amphibians in a variety of habitats at high and low altitudes on or between the Great Dividing Range and the coast. The incidence of chytridiomycosis was higher in winter, with 53% of wild frogs from Queensland and New South Wales dying in July and August. Other diseases were much less common and were detected mostly in spring and summer. In experimental infections, lower temperatures enhanced the pathogenicity of B. dendrobatidis in M. fasciolatus. All 16 frogs exposed to B. dendrobatidis at 17 degrees C and 23 degrees C died, whereas 4 of 8 frogs exposed at 27 degrees C survived. However, the time until death for the frogs that died at 27 degrees C was shorter than at the lower temperatures. Infections in survivors were eliminated by 98 days. CONCLUSION: Chytridiomycosis is a major cause of mortality in free-living and captive amphibians in Australia and mortality rate increases at lower temperatures. PMID- 15354854 TI - Cutaneous leishmaniasis in red kangaroos. PMID- 15354855 TI - Progress towards poliomyelitis eradication in Afghanistan and Pakistan, January 2003 to May 2004. PMID- 15354856 TI - Estrogen receptor alpha gene polymorphisms and bone mineral density in healthy children and young adults. AB - The accretion of peak bone mass is largely under genetic control, and one of the potential candidate genes is the estrogen receptor alpha (ERalpha) gene. The association of ERalpha gene polymorphisms with bone mineral density (BMD) was investigated in a group of 147 healthy caucasian children, adolescents, and young adults (57 boys and 90 girls) in a cross-sectional and longitudinal study. The mean age was 11.3 years (4.3-19.9 years) at baseline and 15.6 years (7.6-25.3 years) at follow-up. Lumbar spine, total body BMD, and body composition were measured by dual energy X-ray absorptiometry and expressed as age- and sex adjusted standard deviation scores (SDS). We analyzed two restriction fragment length polymorphisms, Pvull and Xbal, and haplotypes thereof. Subjects homozygous for haplotype 1 (px) (33% of the population) had 0.4 SD (standard deviation) lower lumbar spine BMD (P = 0.02) and bone mineral apparent density (BMAD) (P = 0.04) than those heterozygous or noncarriers for haplotype 1 (px) at baseline. Analysis of the follow-up data gave similar results. The association was stronger for the prepubertal than for the postpubertal subjects. Vertebral width SDS, total body BMD SDS, height SDS, body mass index SDS, lean body mass SDS, and percentage fat SDS did not significantly differ between the haplotypes. The age of menarche was not related to any of the haplotypes in girls. The present study shows that Pvull-Xbal ERalpha gene polymorphism is associated with BMD during childhood. PMID- 15354857 TI - Estrogen receptor beta dinucleotide (CA) repeat polymorphism is significantly associated with bone mineral density in postmenopausal women. AB - Significant associations between the lengths of a highly polymorphic dinucleotide (CA) repeat located within the human estrogen receptor beta (ESR2) gene on chromosome 14, bone mineral density (BMD) and androgen levels have been reported previously in premenopausal women. We measured the size of this microsatellite repeat in 226 healthy women (60-98 years). After adjustment for age, body mass index, hormone replacement status, and other variables known to influence BMD, women with < 25 CA repeats had significantly higher BMD measured in the total skeleton, lumbar spine, and femoral neck when compared with women having longer alleles. Women with shorter alleles also had higher circulating estrone and estradiol levels that approached statistical significance as compared with women harboring longer alleles after appropriate adjustments were performed in linear regression models. Women having both short and long CA repeats had BMD values in all regions of the skeleton that were midway between those found in women homozygous for longer or shorter repeat sizes. Because the ESR2 CA repeat size was neither associated with change in BMD nor serum levels of biochemical markers of bone turnover, it is likely that ESR2 CA repeat genotype is significantly linked to the attainment of peak bone mass in women. PMID- 15354858 TI - Bone mineral density of the lumbar spine is associated with TNF gene polymorphisms in early postmenopausal Japanese women. AB - We investigated the relationships between tumor necrosis factor (TNF) gene polymorphism, circulating TNF-alpha (TNF-alpha) concentrations, and bone mineral density (BMD) in the lumbar spine. TNF gene polymorphisms studied were the Nco I polymorphism within the first intron of TNF-beta (TNF-beta) and three single nucleotide polymorphisms in the promoter region of the TNF-alpha gene, at positions -857, -863, and -1031. Allelic variants of the TNF gene were identified using restriction fragment length polymorphism (RFLP) analysis in 177 postmenopausal Japanese women within 10 years after menopause, aged 56.4 +/- 4.5 years (mean +/- SD). A significantly higher prevalence of the alleles TNF-alpha 863A (20.3% versus 9.9%) and TNF-alpha-1031C (21.3% versus 12.4%) was seen in the low BMD group (Z-score < 0, n = 91) than in the high BMD group (0 < Z-score, n = 86). In genotype analysis, although difference did not reach a significant level, women with the rarest allelic variants, i.e., homozygous TNFbl, TNF-alpha-863A, and TNF-alpha-1031C, showed the lowest BMD Z-scores. Women with another rarest allelic variant, TNF-alpha-857T/T had significantly lower BMD Z-scores than did women with TNF-alpha-857C/T or -857C/C. The BMD Z-score decreased significantly with an increase in the total number of such rare alleles. Serum concentrations of TNF-alpha did not differ significantly among groups divided by genotypes. Multiple linear regression analysis revealed that the total number of rare alleles, in addition to the body mass index and the number of years since menopause, was an independent predictor of the BMD. These presumptive functional polymorphisms of the TNF gene may be associated with the lumbar spine BMD in early postmenopausal Japanese women. PMID- 15354859 TI - Transforming growth factor-beta1 gene polymorphisms and bone turnover, bone mineral density and fracture risk in southern Chinese women. AB - Genetic contributions play an important role in determining bone mineral density (BMD) and bone turnover. Transforming growth factor-beta (TGF-beta) is abundant in bone and has been implicated as an important regulator of both bone formation and resorption. Several polymorphisms of the TGF-beta1 gene have recently been suggested to be associated with BMD and susceptibility to osteoporotic spine fractures. To determine the relationship between TGF-beta1 polymorphisms and BMD in southern Chinese women, three SNPs at C(-1348) -T, T29 -C, and T(861-20) -C of TGF-beta1 gene were analyzed in 237 postmenopausal southern Chinese women by RFLP and direct sequencing. BMD at the lumbar spine and hip region, biochemical markers of bone turnover, as well as serum levels of TGF-beta1 were measured. Only the T29 -C polymorphism of TGF-beta1 gene was associated with BMD and fracture risk. The prevalence of fragility fractures was significantly higher in individuals with TC genotype (P < 0.05). Serum alkaline phosphatase and osteocalcin levels as well as urinary N-telopeptide excretion were significantly higher in women with TC than with TT or CC genotypes, and the difference remained significant after adjusting for age and BMI (all P < 0.05). Women with TC genotype had lower BMD at the trochanteric (P < 0.03) and total hip region (P = 0.05). No difference was observed in the serum TGF-beta1 levels among the three genotypes. In conclusion, an association between T29 -C polymorphisms of TGF beta1 gene and BMD, bone turnover as well as fragility fractures were demonstrated in postmenopausal southern Chinese women. PMID- 15354860 TI - Interactions of amelogenins with octacalcium phosphate crystal faces are dose dependent. AB - Amelogenins, the major protein components of the enamel extracellular matrix, are postulated to be involved in controlling the elongated and oriented growth of enamel carbonated apatite crystals. In order to clarify the functional role of amelogenin during the early stage of enamel biomineralization, octacalcium phosphate (OCP) crystals, known to be potent precursors of hydroxyapatite, were grown in 1-10% (w/w) native bovine and two recombinant murine amelogenins. Amelogenins were solution-like at 1% and formed gel at 10%, while 5% amelogenins became gel after reaction and it was inhomogeneous and porous. Morphological changes of OCP crystals were evaluated as the function of amelogenin concentration by analyzing the mean values of length, width, thickness, their reduction ratios (L/Lc, W/Wc, T/Tc) as well as L/W and W/T ratios. Length, width, and thickness decreased in a does-dependent manner. Length decreased almost linearly in 1%-10%, whereas width decreased drastically in 1%-5% while the decrease from 5% to 10% was small. As a result, elongated morphology of OCP crystal was most emphasized in 5% bovine amelogenins and rM166 and 2%-5% rM179. The size reduction was in the order of W/Wc < L/Lc < T/Tc. We therefore concluded that amelogenin interaction with crystal faces was in the order (010) > (001) > (100). At all concentrations, W/ Wc was significantly the smallest. This indicated that the primary role of amelogenin was to decrease the width of OCP by blocking the hydrophobic (010) faces. We suggest that the drastic decrease of crystal width is the result of interaction of the densely packed nanospheres in 5%-10% amelogenin. PMID- 15354861 TI - Dentin sialoprotein and phosphoprotein induce neutrophil recruitment: a mechanism dependent on IL-1beta, TNF-beta, and CXC chemokines. AB - Dentin is a reservoir of several potentially active molecules, and dentin sialoprotein (DSP) and dentin phosphoprotein (DPP) are the two major non collagenous proteins. It has been established that dentin molecules are released as a consequence of osteoclast action during the resorption process. Along with osteoclasts, inflammatory cells seem to play an important role at sites of root resorption. Although the role of dentin molecules in dentinogenesis is well known, their role in pathological processes associated with dentin matrix dissolution is unclear. Recent studies have suggested that dentin components may function as chemotactic and activator signals for inflammatory cells at these sites. Herein we present evidence that demineralized dentin crude extract, DSP, and DPP induced doseand time-dependent neutrophil migration into the peritoneal cavity of mice and that this activity was inhibited by dexamethasone, but not by indomethacin or MK886. The blockade of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) receptors inhibited neutrophil accumulation. The neutrophil migration was also diminished in the absence of the chemokines cytokine-induced neutrophil chemoattractant (KC) and macrophage inflammatory protein-2 (MIP-2), but not in the absence of macrophage inflammatory protein 1alpha (MIP-1alpha). These results demonstrate that dentin induces neutrophil migration via the synthesis of IL-1beta, TNF-alpha, and chemokines and they suggest that dentin matrix proteins may have an active role in inflammatory cell recruitment during pathological processes associated with dentin and bone matrix dissolution. PMID- 15354862 TI - Osteoblastic cells express phospholipid receptors and phosphatases and proliferate in response to sphingosine-1-phosphate. AB - Sphingosine-1-phosphate (S1P) is a naturally occurring polar phospholipid that exerts growth-factor--like effects in a number of cell types, potentially by endocrine, paracrine, and intracrine mechanisms. The biological actions of S1P are mediated in part by its specific binding to at least four members of the edg/lp family of G protein-coupled receptors (edg 1, 3, 6, and 8), and further regulated by the actions of specific cell membrane-bound lipid phosphate phosphatases (LPPs). Recent evidence has suggested that S1P may act as an osteoblast growth factor. In the current study, we demonstrate expression in osteoblastic cells of the S1P receptors edg 1, edg 5 and edg 8; the lipid phosphate phosphatases LPP-1, LPP-2, and LPP-3, and the S1P phosphatase mSPP1. S1P exerts dose-dependent mitogenic effects in cultures of primary rat osteoblastic cells and SaOS-2 cells. S1P also activates p42/44 mitogen-activated protein (MAP) kinases in osteoblastic cells, and the proliferative effects of S1P in these cultures are partially abrogated by PD-98059 and U-0126, specific inhibitors of p42/44 MAP kinase signaling. S1P-induced p42/44 MAP kinase activation in osteoblastic cells is dependent on functional Gi proteins and intracellular calcium fluxes, but not on protein kinase A, phospha-tidylinositol 3 kinase, or protein Kinase C. These data demonstrate (1) the expression by osteoblastic cells of several cell membrane-bound components of phospholipid signaling and (2) that S1P is a potent osteoblast mitogen, the proliferative action of which is mediated by a signaling pathway that involves Gi proteins, intracellular calcium, and p42/44 MAP kinases. PMID- 15354863 TI - Improving metabolic control reverses the histomorphometric and biomechanical abnormalities of an experimentally induced bone defect in spontaneously diabetic rats. AB - Insulin-dependent type 1 diabetes mellitus (IDDM) has been shown to alter the properties of bone and to impair fracture-healing in both humans and animals. The objective of this study was to examine changes in the histomorphometric and mechanical parameters of bone and remodeling during bone-defect healing, depending on the diabetic metabolic state in spontaneously diabetic BB/O(ttawa)K(arlsburg) rats, a rat strain that represents a close homology to IDDM in humans. A standardized bone-defect model was chosen and based on blood glucose values at the time of surgery (mg%), postoperative blood-glucose course (mg%), and postoperative insulin requirements (IU/kg). A total of 120 spontaneously diabetic BB/OK rats were divided into groups with a well compensated (n = 60; 169 +/- 102 mg%; 230 +/- 126 mg%; and 2.2 +/- 1.1 IU/ kg) or poorly compensated (n = 60; 380 +/- 159 mg%; 359 +/- 89 mg%; and 5.4 +/- 1.1 IU/kg) metabolic state. Sixty LEW.1A rats served as the normoglycemic controls (93 +/- 19 mg%). Fifteen animals from each group were killed on postoperative days 7, 14, 24, and 42, and specimens were processed undecalcified for quantitative bone histomorphometry and for biomechanical testing. Our study showed in terms of bone histomorphometry, within the first 14 days, that severe mineralization disorders occurred exclusively in the rats with a poorly compensated diabetic metabolic state with a highly significant (P < 0.001) or significant (P < 0.01) decrease of all fluorochrome-based parameters of mineralization, apposition, formation and timing of mineralization, as well as significantly decreased values of biomechanical properties (P < 0.05) in comparison to the spontaneously diabetic rats with a well-compensated metabolic state and to the control rats. Bone-defect healing in spontaneously diabetic BB/ OK rats is retarded exclusively in a poorly compensated diabetic metabolic state. This study suggests that strictly controlled insulin treatment resulting in a well-compensated diabetic metabolic state will ameliorate the impaired early and late parameters of IDDM bone-defect healing. PMID- 15354864 TI - Tbx2 represses expression of Connexin43 in osteoblastic-like cells. AB - Tbx2 belongs to a family of developmental transcription regulatory factors. We evaluated whether the gap junction protein Connexin43 (Cx43), an important regulator of osteoblast function and bone development, may be a downstream target gene regulated by Tbx2. The Cx43 promoter contains direct repeats of the consensus T-box binding motif, TCACAC, and moreover, Tbx2 and Cx43 show overlapping expression domains in precursors to bone and in osteoblasts. In vitro analysis showed that the Cx43 promoter contains two Tbx2 binding sites, and this binding was dependent on the TCACAC consensus sequence. Transient transfection analysis with a Cx43 promoter-driven lacZ reporter construct revealed negative regulation mediated by these two Tbx2 binding sites in osteoblast-like cells. Thus, downregulation of Tbx2 led to de-repression of wild-type Cx43 promoter activity, whereas a promoter construct with mutated binding sites showed no de repression. In stably transfected osteosarcoma cells in which expression of the endogenous Tbx2 gene was downregulated with a Tbx2 antisense construct, a marked de-repression of the endogenous Cx43 gene was observed. This was accompanied by a marked increase in the abundance of Cx43 gap junctions and increased functional gap junction-mediated cell-cell communication. Analysis of lacZ expression in transgenic mice containing the mutated Cx43 promoter-driven lacZ construct further suggested de-repression of the Cx43 promoter in limb buds, a region destined to give rise to long bones of the limbs. Taken together, these findings indicate that the promoter of Cx43 is repressible by Tbx2, both in cultured osteoblast-like cells in vitro and likely in the developing embryo. PMID- 15354865 TI - The role of PrP in health and disease. AB - Transmissible spongiform encephalopathies (TSEs) such as scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle or Creutzfeldt-Jacob disease (CJD) and Gerstmann-Straussler-Scheinker syndrome (GSS) in humans, are caused by an infectious agent designated prion. The "protein only" hypothesis states that the prion consists partly or entirely of a conformational isoform of the normal host protein PrPc and that the abnormal conformer, when introduced into the organism, causes the conversion of PrPc into a likeness of itself. Since the proposal of the "protein only" hypothesis more than three decades ago, cloning of the PrP gene, studies on PrP knockout mice and on mice transgenic for mutant PrP genes allowed deep insights into prion biology. Reverse genetics on PrP knockout mice containing modified PrP transgenes was used to address a variety of problems: mapping PrP regions required for prion replication, studying PrP mutations affecting the species barrier, modeling familial forms of human prion disease, analysing the cell specificity of prion propagation and investigating the physiological role of PrP by structure-function studies. Many questions regarding the role of PrP in susceptibility to prions have been elucidated, however the physiological role of PrP and the pathological mechanisms of neurodegeneration in prion diseases are still elusive. PMID- 15354866 TI - The peripheral nervous system and the pathogenesis of prion diseases. AB - Prion diseases are inevitably fatal neurodegenerative conditions which affect humans and a wide variety of animals. Unlike other protein aggregation diseases such as Alzheimer's, Parkinson's, and polyglutamine repeat diseases, prion diseases are unique in that they are transmissible. Therefore, prion diseases are also called transmissible spongiform encephalopathies. A number of prion diseases are caused by peripheral uptake of the infectious agent. In order to reach their target, the central nervous system, prions enter their host, accumulate and replicate in lymphoid organs, and eventually spread to the central nervous system via peripheral nerves. Once the agent has reached the central nervous system, disease progression is rapid, resulting in neurodegeneration and death. In this article, we review the state of knowledge on the routes of neuroinvasion used by the infectious agent in order to gain access to the central nervous system upon entry into extracerebral sites. PMID- 15354867 TI - Impacts and concerns for vCJD in blood transfusion: current status. AB - The impact of vCJD upon blood transfusion practice hinges on its lymphoreticular involvement. B lymphocytes play a key supporting role for the capture and replication of infectivity by follicular dendritic cells of the lymphoid tissue in animal models of transmissible spongiform encephalopathies (TSE) and tonsils, spleen and appendix in man can harbour vCJD infectivity, a situation not seen with the other human TSEs. Leucodepletion of blood donations in the UK was implemented to reduce possible vCJD transmission and preliminary data suggests that white cell associated infectivity will be effectively removed although plasma infectivity will not. Blood screening assays are under development but none yet are ready for application. The conformation dependant immunoassay, based on differences in secondary and tertiary structure between normal and TSE associated abnormal prion protein, has a sensitivity now approaching the best bioassay. Even so further development is needed to detect the fg/ml levels likely in the event that vCJD blood does contain abnormal prion, which is as yet unproven. Surrogate assays, such as for erythroid associated factor, may provide additional means of identifying donors harbouring vCJD. Validation of clearance of TSEs from pooled plasma products consistently demonstrates effective removal of the agents in downscaled systems and studies comparing vCJD, BSE and scrapie agents yield similar results. Many approaches to therapy are under investigation, in cell culture and animal models, targeted to normal or abnormal prion metabolism, including chemical and immunological interventions. Efficacy of quinacrine/chlorpromazine and pentosan polysulphate in a clinical setting, and agents yet to be used, will be more accurately known following recent agreement of clinical drug evaluation protocols. PMID- 15354868 TI - Genetic studies in relation to kuru: an overview. AB - Kuru is a subacute neurodegenerative disease presenting with limb ataxia, dysarthria, and a shivering tremor. The disease progress to complete motor and mental incapacity and death within 6 to 24 months. Neuropathologically, a typical pattern of neuronal loss, astrocytic and microglial proliferation, characteristic "kuru-type" amyloid plaques, and PrP deposits in the cerebral cortex and cerebellum are observed. Kuru is the prototype of a group of human transmissible spongiform encephalopathies (TSEs), or "prion" diseases, that include hereditary, sporadic and infectious forms. The latest member of this group, the variant Creutzfeldt-Jakob disease (vCJD), linked to transmission of bovine spongiform encephalopathy (BSE) to humans, shows features similar to kuru. Kuru has emerged at the beginning of the 1900s in a small indigenous population of New-Guinean Eastern Highlands, reached epidemic proportions in the mid-1950s and disappeared progressively in the latter half of the century to complete absence at the end of the 1990s. Early studies made infection, the first etiologic assumption, seem unlikely and led to a hypothesis that kuru might be a genetically determined or genetically mediated illness. After transmissibility of kuru had been discovered and all major epidemiologic phenomena adequately explained by the spread of an infectious agent with long incubation period through the practice of cannibalism, the pattern of occurrence still continued to suggest a role for genetic predisposition. Recent studies indicate that individuals homozygous for Methionine at a polymorphic position 129 of the prion protein were preferentially affected during the kuru epidemic. The carriers of the alternative 129Met/Val and 129Val/Val genotypes had a longer incubation period and thus developed disease at a later age and at a later stage of the epidemic. Observations made during the kuru epidemic are helpful in the understanding of the current vCJD outbreak, and vice versa clinical and experimental data accumulated in studies of other TSE disorders contribute to better understanding of the documented kuru phenomena. PMID- 15354869 TI - The genetics of scrapie in sheep and goats. AB - Scrapie, an invariably fatal disease of sheep and goats, is a transmissible spongiform encephalopathy (TSE). The putative infectious agent is the host encoded prion protein, PrP. The development of scrapie is closely linked to polymorphisms in the host PrP gene. The pathogenesis of most TSEs involves conversion of normal, cellular PrP into a protease-resistant, pathogenic isoform called PrPSc. The conversion to PrPSc involves change in secondary structure; it is impacts on these structural changes that may link polymorphisms to disease. Within the structured C-terminal part of PrP polymorphisms have been reported at 15 and 10 codons of the sheep and goat PrP genes respectively. Three polymorphisms in sheep are acutely linked to the occurrence of scrapie: A136V, R154H and Q171R/H. These generate five commonly observed alleles: ARQ, ARR, AHQ, ARH and VRQ. ARR and AHQ are associated with resistance; ARQ, ARH and VRQ are associated with susceptibility. There are subtle effects of specific allele pairings (genotypes). Generally, more susceptible genotypes have younger ages at death from scrapie. Different strains of scrapie occur which may attack genotypes differently. Different sheep breeds vary in the assortment of the five alleles that they predominantly encode. The reason for this variation is not known. Furthermore, certain genotypes may be susceptible to scrapie in some breeds and resistant in others. The explanation is not known, but may relate to different scrapie strains circulating in different breeds, or there may be effects of other genes which modulate the effect of PrP. PMID- 15354870 TI - Molecular mechanisms of neurotoxicity of pathological prion protein. AB - Transmissible Spongiform Encephalopathies or prion related disorders are fatal and infectious neurodegenerative diseases characterized by extensive neuronal apoptosis and accumulation of a misfolded form of the cellular prion protein (PrP), denoted PrP(Sc). Although the mechanism of neurodegeneration and the involvement of PrP(Sc) is far from clear, data indicates that neuronal apoptosis might be related to activation of several signaling pathways, including proteasome dysfunction, alterations in prion maturation pathway and endoplasmic reticulum (ER) stress. In this article we describe recent studies investigating the molecular mechanism of PrP(Sc) neurotoxicity. We propose a model in which the key step in the pathogenesis of prion disorders, independent on their etiology, is the alteration of ER-homeostasis due to drastic modifications of the physicochemical properties of PrP, leading to the activation of ER-dependent signaling pathways that controls cellular survival. PMID- 15354871 TI - Sphingolipids in inflammation: roles and implications. AB - Sphingolipids, historically described as potential reservoirs for bioactive lipids, presently define a new family of cellular mediators, joining the well established glycerolipid-derived mediators of signal transduction such as diacylglycerol, phosphatidylinositides, and eicosanoids. Sphingolipid metabolism is clearly involved in the regulation of cell growth, differentiation, and programmed cell death. Indeed, a majority of the greater than four thousand studies conducted on sphingolipids during the past five years were investigations of the role of sphingolipids as cellular bioregulators. Studies spanning more than a decade have shown multiple interactions and intersections of the sphingolipid-mediated pathways and the eicosanoid pathway. This review will discuss the emerging mechanisms by which sphingolipids induce inflammatory responses via the eicosanoid pathway in addition to linking previous literature on sphingolipids and inflammation with newer findings of distinct roles for sphingosine-1-phosphate in regulating cyclooygenase-2 and ceramide-1-phosphate in the regulation of cytosolic phospholipase A2alpha. Finally, the relationship between bioactive sphingolipids and inflammation is discussed. PMID- 15354872 TI - Fluorescence molecular imaging of small animal tumor models. AB - In vivo imaging of molecular events in small animals has great potential to impact basic science and drug development. For this reason, several imaging technologies have been adapted to small animal research, including X-ray, magnetic resonance, and radioisotope imaging. Despite this plethora of visualization techniques, fluorescence imaging is emerging as an important alternative because of its operational simplicity, safety, and cost effectiveness. Fluorescence imaging has recently become particularly interesting because of advances in fluorescent probe technology, including targeted fluorochromes as well as fluorescent "switches" sensitive to specific biochemical events. While past biological investigations using fluorescence have focused on microscopic examination of ex vivo, in vitro, or intravital specimens, techniques for macroscopic fluorescence imaging are now emerging for in vivo molecular imaging applications. This review illuminates fluorescence imaging technologies that hold promise for small animal imaging. In particular we focus on planar illumination techniques, also known as Fluorescence Reflectance Imaging (FRI), and discuss its performance and current use. We then discuss fluorescence molecular tomography (FMT), an evolving technique for quantitative three dimensional imaging of fluorescence in vivo. This technique offers the promise of non-invasively quantifying and visualizing specific molecular activity in living subjects in three dimensions. PMID- 15354873 TI - Trafficking of natural killer cells. AB - Natural killer (NK) cells comprise a set of lymphocytes that is capable of mediating innate immune responses to viral infections, malignancies, and allogeneic bone marrow grafts. This review summarizes what is known about the mechanisms NK cells use to arrive at their sites of action. NK cells express a wide array of adhesion molecules including alphaLbeta2, alphaMbeta2, alphaXbeta2, and alpha4beta1 integrins, ICAM-1, PSGL-1, and L-selectin. Like other immune and inflammatory cells, NK cells use the blood circulation to enter tissues and organs, which requires that they interact with the vessel wall under flow conditions, arrest, and transmigrate. NK cells are able to chemotax to a variety of cytokines and chemokines, including IL-12, IFN-(alpha/beta, CCL2, 3, 4, 5, 7, 8, CXCL8, and CX3CL1. In many cases, NK cells appear to migrate towards these soluble factors without any kind of priming. These cells also appear to distribute in secondary and tertiary lymphoid sites (i.e., spleen, bone marrow, liver, lung, and lymph nodes) both with and without stimulation. In addition to their ability to move throughout the body in an unprimed state, activated NK cells may have increased specificity in homing to sites of inflammation. NK cells not only react to, but also produce IFN-gamma, TNF-alpha, GM-CSF, CCL3, CCL4, and CCL5, enabling them to recruit various immune cells to sites of immune response. PMID- 15354874 TI - Differential TNF-signaling in chronic inflammatory disorders. AB - TNF-alpha is a pleiotropic cytokine with strong proinflammatory and immunomodulatory properties. TNF-alpha plays a critical role in many acute or chronic inflammatory diseases and anti-TNF-strategies have proven to be clinically effective. Two TNF-specific cell surface receptors TNF-R1 and TNF-R2 have been identified and the function of these receptors and the downstream intracellular signal transduction pathways have been extensively studied in vitro. For a long time TNF-R1 was considered to be the predominant mediator of TNF-signaling, whereas TNF-R2 was ascribed only auxilliary function. However, there is increasing clinical and experimental evidence for an important independent role of p80 signaling in chronic inflammatory conditions. It is conceivable that the multiple TNF-mediated chronic inflammatory disorders differ in terms of the ligand form (soluble TNF-alpha versus membrane bound TNF-alpha), the receptor (TNF-R1 versus TNF-R2) and the downstream signaling cascades utilized. The elucidation of the specific characteristics of TNF-signaling in distinct inflammatory disorders will lead to a better understanding ot the pathogenesis of these diseases and will be the basis for the development of more specific and more efficient therapeutic approaches. PMID- 15354875 TI - [The thyroid impaired only latently. When to treat, when not to?]. PMID- 15354876 TI - [In spite of lacking abdominal complaints. Intestinal disease was diagnosed because of the lips]. PMID- 15354877 TI - [High incidence and resistance rates in Eastern Europe. Is tuberculosis coming back?]. PMID- 15354878 TI - [The family doctor as professional travel counselor]. PMID- 15354879 TI - [Humanopathogenic parasites--an unwelcome import]. AB - Humanopathogenic parasites represent a health risk for travelers to tropical countries. They can be picked up by direct contact, via contaminated food and drink, the bites of blood-sucking insects and leeches, as also via direct penetration through the skin. Not only the bite and sting wounds themselves are unpleasant. The true danger to the victim is represented, in particular, by the bacteria, viruses or parasites that are transmitted in this way. They can give rise to protracted diseases such as hepatitis B, cutaneous or intestinal disorders. The most effective preventive measure is exposure prophylaxis. PMID- 15354880 TI - [The elderly traveler]. AB - In recent years, the number of senior citizen travelers has been increasing. As a result of this trend, the general practitioner is every more frequently being confronted with the problem of counseling his elderly patients. A particularly important point is that underlying illnesses and long-term medication must be adapted to the destination, traveling time and style of traveling. Since considerable immunization deficits can be found among senior citizens, the application of prophylactic vaccination prior to the jou rney presents an ideal opportunity to bring the patient's immunization status up to date in terms of generally recommended vaccinations. Should chemoprophylaxis be necessary, consideration must always be given to drug interactions. By making appropriate recommendations in terms of health-related and destination-related dos and don'ts, the risk of developing health problems while traveling can be minimized. PMID- 15354881 TI - [Diarrhea in tropical countries--what is to be done?]. AB - Diarrhea continues to be the most common cause of health problems encountered by travelers to tropical or subtropical countries. Every second traveler is affected. Since traveler's diarrhea cannot be reliably prevented by prophylactic measures, the sole option is self-medication. To accomplish this, comprehensive counseling of the traveler is a must. Appropriate treatment properly implemented can be highly effective and well tolerated and may curtail the illness and prevent complications. PMID- 15354883 TI - [Danger approaching--insect bites can cause allergic reactions]. PMID- 15354882 TI - [Malaria: prophylaxis, treatment, stand-by medication. A single mosquito bite suffices]. PMID- 15354884 TI - [Suicides peak in May and June. Is the decision to kill oneself dependent on the weather?]. PMID- 15354885 TI - [Travel counseling: enrolling, organization, accounting. Why does colleague Anderten permit to bring along sand]. PMID- 15354886 TI - [Hospital offers fees to physicians. Beware, that can end!]. PMID- 15354887 TI - [A rare skin rash]. PMID- 15354888 TI - [Child with atopy]. PMID- 15354889 TI - A double-edged sword: insurance A to Z. PMID- 15354890 TI - CT scans. PMID- 15354891 TI - Periodontal standard. PMID- 15354892 TI - Oral physicians 'now'! PMID- 15354893 TI - Brand names. PMID- 15354894 TI - More about brand names. PMID- 15354895 TI - Practice management. PMID- 15354896 TI - Radiography and low birth weight. PMID- 15354897 TI - The vaccine controversy. PMID- 15354898 TI - The effect of esthetic consultation methods on acceptance of diastema-closure treatment plan: a pilot study. AB - BACKGROUND: The authors conducted a study to determine which of four consultation methods helped patients best understand a proposed treatment plan for maxillary anterior diastema closure. METHODS: The authors presented 24 subjects with four types of consultation in random order: before-and-after photographs of other patients, diagnostic models with wax setups, resin-based composite/esthetic preview/mock-ups and computer-imaging simulations. After viewing each method, the authors asked the subjects about treatment acceptability. At the end of the demonstrations of all four methods, the authors asked the subjects which consultation method helped them best understand the proposed treatment plan. RESULTS: A total of 87.5 percent of the subjects indicated that they would accept the proposed treatment plan after they were shown the computer-imaging simulation, 50 percent said they would after they were shown the resin-based composite/esthetic preview/mock-up, 41.7 percent said they would after they were shown photographs of other patients, and 25 percent said they would after they were shown diagnostic models with wax setups. When asked which method helped them best understand the proposed treatment, 54.2 percent of the subjects selected computer-imaging simulation, 33.3 percent selected resin-based composite/esthetic preview/mock-ups, and 12.5 percent selected before-and-after photographs of other patients. None of the subjects selected diagnostic models with wax setups. A chi2 test for goodness of fit indicated that these differences were statistically significant. CONCLUSION: Subjects preferred computer-imaging simulation to the other three consultation methods, and they indicated that computer-imaging simulation provided a better understanding of the proposed treatment plan for diastema closure. CLINICAL IMPLICATIONS: The use of computer-imaging simulation enhances the patient's understanding of a proposed treatment plan concerning maxillary anterior diastema closure. PMID- 15354899 TI - Risk indicators for posterior tooth fracture. AB - BACKGROUND: Identifying posterior teeth that are at heightened risk of developing cusp fracture is an inexact science. Risk indicators based on controlled observations are not available, and dentists' assessments vary. METHODS: The authors conducted a case-control study of cusp fracture in restored posterior teeth. They evaluated 39 potential risk indicators identified in previous uncontrolled studies for an association with fracture in 200 patients with fractures and 252 patients without fractures. These risk indicators delineated patients' clinical characteristics and behaviors, as well as clinical characteristics of individual teeth. The authors used logistic regression to develop models identifying risk indicators associated with fracture, both between case and control subjects and between case and comparison teeth in case subjects. RESULTS: Two risk indicators appeared in both models. The presence of a fracture line and an increase in the proportion of the volume of the natural tooth crown occupied by the restoration substantially increased the odds of fracture (P < .001). Additional risk indicators were unique to the case subject-control subject model, including subject age and other measures related to the relative size of the restoration or to loss of dentinal support. Neither patient behaviors such as clenching, grinding and biting hard objects nor occlusal characteristics such as guidance, cusp anatomy and general wear patterns were strong predictors of fracture risk. CONCLUSIONS: Among posterior teeth with restorations, two clinical features were strongly associated with the risk of cusp fracture: presence of a fracture line in the enamel and proportional volume of the restoration. CLINICAL IMPLICATIONS: Dentists assessing the risk of fracture should consider a detectable fracture line or a high ratio of restoration-to-total-crown volume as important indicators of elevated risk. PMID- 15354900 TI - The effect of saliva contamination on enamel and dentin using a self-etching adhesive. AB - BACKGROUND: The authors conducted a study to evaluate the effect of saliva contamination on enamel and dentin before or after application of a self-etching adhesive. METHODS: The authors divided 120 extracted molars into enamel and dentin groups of 60 specimens each, and then further divided each group into three groups of 20 specimens each. Resin-based composite cylinders were bonded to tooth specimens that were uncontaminated (control) or were contaminated with saliva either before or after application of the adhesive and then loaded to the failure point. The authors calculated the mean shear bond strength for each group and subjected data to a one-way analysis of variance, or ANOVA, to test for differences among groups. RESULTS: Mean shear bond strength was 16.7 megapascals (+/- 3.0 standard deviation, or SD) for the dentin control group, 14.8 MPa (+/- 2.3 SD) for dentin contaminated with saliva before adhesive application, 15.9 MPa (+/- 3.8 SD) for dentin contaminated with saliva after adhesive application, 19.1 MPa (+/- 4.8 SD) for the enamel control group, 12.3 MPa (+/- 4.4 SD) for enamel contaminated with saliva before adhesive application, and 13.1 MPa (+/- 5.5 SD) for enamel contaminated with saliva after adhesive application. One-way ANOVA detected no difference in the dentin groups, but the authors found a significant difference among enamel groups. Uncontaminated enamel had higher bond strengths than either of the contaminated enamel groups. CONCLUSIONS: Saliva contamination did not affect the dentin shear bond strength of the self-etching adhesive. Saliva did have a detrimental effect on enamel bond strength. CLINICAL IMPLICATIONS: Newer generations of adhesives appear to be more tolerant of saliva contamination, but proper isolation still should be accomplished until further research proves otherwise. PMID- 15354901 TI - The impact of post space preparation with Gates-Glidden drills on residual dentin thickness in distal roots of mandibular molars. AB - BACKGROUND: Posts frequently are used to retain buildups. The authors examined the effects of post space preparation with Gates-Glidden drills on residual dentin thickness in distal roots of mandibular molars. METHODS: The authors embedded 26 root-treated mandibular molars in endodontic cubes and sectioned them perpendicularly to their long axis at 1.8-millimeter intervals. They placed Gates Glidden drills nos. 3, 4, 5 and 6 sequentially to 5 mm short of working length in the distal canals. After using each drill size, the authors measured the dentin thickness from light microscopic images of the tooth slices. RESULTS: The authors analyzed data to detect changes resulting from canal instrumentation. They performed statistical analysis by partitioning the variability in a nested analysis of variance. Strip perforations occurred with a no. 4 Gates-Glidden drill 7.3 percent of the time and more frequently with larger drills. After endodontic treatment alone, the canal wall on the furcal side was less than 1 mm thick 82 percent of the time and less than 0.5 mm thick 17.5 percent of the time. CONCLUSIONS: Dentin thickness correlates inversely to post space diameter. A no. 4 Gates-Glidden drill caused strip perforations in 7.3 percent of the canals studied, and therefore the authors recommend that Gates-Glidden drills larger than a no. 3 not be used in these roots. After endodontic treatment, the furcation-side dentin thickness was less than 1 mm in 82 percent of the teeth. CLINICAL IMPLICATIONS: Post space preparation in mandibular molars carries significant risk of perforation. Post space in such teeth should be limited to the endodontically prepared canal. PMID- 15354902 TI - Tooth erosion caused by chewing aspirin. AB - BACKGROUND: Although the effects of aspirin on the oral mucosa are well documented, there is little documentation of the effects of aspirin-chewing on the enamel and dentin. CASE DESCRIPTION: The authors present two cases of patients with damage to their tooth enamel and dentin. Both patients had similiar symptoms, but had not been told that chewing aspirin could harm tooth structure. The authors identify clinical signs and symptoms and discuss ways to prevent erosion. CLINICAL IMPLICATIONS: The common factor in these cases is that aspirin was the only possible cause of the tooth erosion. Dentists should be aware of the effects of aspirin-chewing on tooth structure and advise their patients accordingly. PMID- 15354903 TI - Permanent cementation of crowns on teeth with minimal tooth structure. PMID- 15354904 TI - Continuing education: the good, the bad and the ugly. AB - Dental CE is mandatory for ongoing competency in dental practice. There are numerous methods of obtaining CE, some of which are highly successful and others that are of minimal value. In this article, I have described and prioritized the various methods of obtaining CE according to their effectiveness as I perceive it. CE in dentistry can be enjoyable, will allow dentists to serve their patients at a higher level of competency and will keep practitioners mentally stimulated as they learn new concepts and techniques. PMID- 15354905 TI - Essential questions regarding data, backups and online backups. AB - BACKGROUND: As dental offices switch from traditional paper records to electronic records, data safety becomes a concern. Simple safety measures such as off-site data backup should be considered by dental offices that have electronic records. CONCLUSIONS: Loss of practice data such as patient charts, financial records and the appointment schedule could result in serious damage to a dental practice. Electronic computer records can be safeguarded using a backup system. There are several acceptable backup systems that have different benefits and shortcomings. Online backup is the newest backup method, and it offers some significant improvements over older systems. PRACTICE IMPLICATIONS: By understanding why data need to be backed up, what data need to be backed up and how to back them up, dental practices can protect their practice data properly. Failure to follow proper backup procedures could result in significant loss. PMID- 15354906 TI - Dental visits among Hispanics in the United States, 1999. AB - BACKGROUND: This article describes dental visits among Hispanics and Hispanic subgroups in the United States. METHODS: This study is based on an analysis of data regarding dental visits among Hispanics two years of age and older from the National Health Interview Survey of 1999. The authors compared the 1999 data with a combined sample from 1978 through 1980. RESULTS: Although the level of dental visits for all Hispanic subgroups increased between the period of 1978 through 1980 and 1999, the level of dental visits was lowest among Mexican-Americans. For Mexican-Americans born outside the United States, the longer they had lived in the United States, the more likely they were to have visited a dentist. CONCLUSIONS: Increases in the utilization of dental services among Hispanics provide some optimism for reductions in the level of untreated oral disease among Hispanics in the future. However, if increases in dental care utilization among Mexican-Americans fail to keep pace with those among other segments of the U.S. population, this population group risks falling further behind. PRACTICE IMPLICATIONS: Hispanic patients will make up a greater percentage of all dental patients in the future. The impact of this growth will vary greatly by state. PMID- 15354907 TI - Collections and financial controls. AB - There is a reason why most businesses manage fees and money very closely with specific guidelines. It is not to be inflexible, but rather to establish excellent financial controls to ensure that the financial health of the business (or practice) remains strong while achieving outstanding customer (or patient) satisfaction. PMID- 15354908 TI - The effect of a triclosan/copolymer/fluoride liquid dentifrice on interproximal enamel remineralization and fluoride uptake. AB - BACKGROUND: The authors measured the enamel remineralization of dental interproximal surfaces by a triclosan/copolymer/fluoride liquid dentifrice to test its performance against that of a standard toothpaste. METHODS: In a randomized, crossover, blinded, in situ protocol, the authors fitted 19 healthy adult subjects with mandibular appliances holding bilateral, interproximal bovine enamel slabs. They measured initial abrasion levels and fluoride content. In a two-phase crossover protocol, they evaluated the percentage of surface mineral recovery, or SMR, and fluoride uptake caused by the experimental dentifrice and a fluoride-containing traditional control toothpaste. RESULTS: Abrasion depths were similar and not different statistically (P > .05). The interproximal blocks exposed to the triclosan-containing liquid dentifrice had a mean fluoride uptake that was nearly 100 parts per million (13.1 percent) greater than that achieved with the American Dental Association-accepted control dentifrice. This finding, while not statistically significant, may indicate enhanced performance of the experimental dentifrice because the initial enamel slab abrasion depths were shown to be quite uniform. Moreover, the blocks exposed to the experimental toothpaste had a 49.8 percent SMR, while the positive control blocks had an SMR of only 36.9 percent. This enhanced remineralization performance was statistically significant (P < .05). CONCLUSIONS: This is the first study to show enhanced performance of a triclosan-containing liquid dentifrice in preventing interproximal dental caries. The difference in percentage of SMR was statistically significant and is likely to be clinically relevant. CLINICAL IMPLICATION: The results of this preliminary study offer evidence supporting clinicians' recommendation of the new liquid dentifrice to patients who are prone to developing interproximal carious lesions. PMID- 15354909 TI - Stress, strain and unpaid claims. PMID- 15354910 TI - For the dental patient. The importance of using mouthguards. Tips for keeping your smile safe. PMID- 15354911 TI - HCQIA immunity: one win and one loss. PMID- 15354912 TI - A comparison of endotracheal tube cuff pressures using estimation techniques and direct intracuff measurement. PMID- 15354913 TI - The power of words. PMID- 15354914 TI - Anesthetic management of a difficult airway in a patient with epidermolysis bullosa: a case report. AB - Epidermolysis bullosa is an inherited skin disease that leads to an array of medical problems. Patients are susceptible to blistering and scar formation following even minor trauma. These patients may present with scarring, limiting the range of motion of their temporal mandibular joint. This case report describes a 15-year-old patient with epidermolysis bullosa presenting for contracture release, with a difficult airway. PMID- 15354915 TI - Arterial cannulation: a critical review. AB - Arterial catheterization for hemodynamic monitoring is used widely in clinical management. Complications of connulation have been recognized since introduction of the technique. This review examines radial, brachial, axillary, and femoral cannulation sites. Waveform distortion, adjacent structure injury, and the incidence of thrombus are described. Computerized subject heading searches were executed using CINAHL and MEDLINE databases. Searches encompassed English language, randomized, controlled trials, reviews, practice guidelines, and meta analyses published from January 1997 to February 2002. Additional studies were identified via review of retrieved literature. Radial cannulation is subject to inaccuracy and thrombus formation, although a benefit is dual circulation. The brachial site is subject to inaccuracy, lacks collateral circulation, and is associated with median nerve injury. Axillary cannulation provides data closely approximating aortic pressure and poses minimal thrombotic risk but is associated with brachial plexus compression. Femoral cannulation provides a pulse contour approximating aortic with minimal thrombotic risk. There is little evidence to show increased incidence of catheter-related systemic infection at this site. PMID- 15354916 TI - The addition of clonidine to bupivacaine in combined femoral-sciatic nerve block for anterior cruciate ligament reconstruction. AB - Clonidine has been shown to prolong sensory analgesia when given as an adjunct to peripheral nerve blocks but has not been evaluated when given in conjunction with a femoral-sciatic nerve block. The purpose of this investigation was to determine whether the addition of clonidine to a femoral-sciatic nerve block would prolong the duration of sensory analgesia in groups of patients undergoing anterior cruciate ligament (ACL) reconstruction. This prospective, randomized, double blind investigation was performed on 64 subjects undergoing ACL reconstruction. Patients were assigned randomly to receive a femoral-sciatic nerve block using 30 mL of 0.5% bupivacaine with 1:200,000 epinephrine (control group) or 30 mL of 0.5% bupivacaine with 1:200,000 epinephrine and 1 microg/kg of clonidine (experimental group). Variables measured included demographics, timed pain intensity measurements, postoperative analgesic consumption, duration of analgesia, and patient satisfaction. No significant differences were noted between groups for pain intensity scores, duration of sensory analgesia, postoperative analgesic requirements, or overall patient satisfaction. Both groups reported minimal amounts of postoperative pain and high analgesic satisfaction scores. Based on our results, we do not recommend the addition of clonidine to a femoral-sciatic nerve block when given to facilitate postoperative analgesia in patients undergoing ACL reconstruction. PMID- 15354917 TI - Childhood airway manifestations of lymphangioma: a case report. AB - Lymphangioma is a congenital malformation of the lymphatic system, often involving areas of the head and neck. The involved structures may include enlarged tongue and lips, swelling of the floor of the mouth, and direct involvement of the upper respiratory tract. The definitive treatment for lymphangioma is surgery, often during the first years of life. Despite surgical removal, lymphangioma may persist. Anesthetic concerns include bleeding, difficulty visualizing the airway, extrinsic and intrinsic pressure on the airway causing distortion, and enlarged upper respiratory structures, including the lips, tongue, and epiglottis. This is a case report of a 9-year-old patient with lymphangioma who had impacted teeth and a suspected odontogenic cyst. There seems to be little information on the optimal anesthetic management for this age group. The challenges with airway management, including bleeding, laryngospasm, and a difficult intubation, are outlined. Awareness of potential airway involvement and possible complications is necessary to provide a safe anesthetic to a patient with lymphangioma. A review of the literature, airway management techniques, and current airway equipment will be discussed. PMID- 15354918 TI - Visual loss as a complication of nonophthalmologic surgery: a review of the literature. AB - Decreased visual acuity and loss of visual ability are devastating anesthetic and surgical complications. The incidence is greater in patients with preexisting hypertension, diabetes, sickle cell anemia, renal failure, gastrointestinal ulcer, narrow-angle glaucoma, vascular occlusive disease, cardiac disease, arteriosclerosis, polycythemia vera, and collagen vascular disorders. Precipitating factors for ischemic optic neuropathy include prolonged hypotension, anemia, surgery, trauma, gastrointestinal bleeding, hemorrhage, shock, prone position, direct pressure on the globe, and long operative times. Prone and Trendelenburg positions can lead to visual loss related to decreased venous return from the head. Visual impairment may result from increased intracranial pressure, which contributes to undue pressure on the optic nerve. The prone position increases the risk of direct compression injury to the orbit and corneal abrasion. Astute attention to positioning is imperative, especially with the prone position. At-risk patients should receive transfusion once the calculated allowable blood loss has been surpassed. Unacceptable hemoglobin and hematocrit values should be corrected preoperatively and levels monitored during the case to avoid intraoperative anemia in at-risk patients. The blood pressure of patients with predisposing diseases should be kept within normal limits. To avoid this devastating complication, it is imperative that anesthesia providers understand contributing factors and prevention strategies. PMID- 15354920 TI - "How do you know that?". PMID- 15354919 TI - Eliminating surgical fires: a team approach. AB - The incidence of surgical fires is rising. One agency estimates that the frequency of surgical fires is comparable to that of wrong-site surgery with an incidence of 50 to 100 fires annually. Anesthetists must assist in the development and implementation of processes that eliminate the disfiguring or deadly results from surgical fires. Understanding the 3 components of fires--heat sources, fuels, and oxidizers--is essential to reduce or eliminate the risk of surgical fires. This AANA Journal course discusses the need for a clearly formulated plan, rehearsing a response to fires in or on the patient, and clearly designated prevention parameters. As part of the surgical team, the vigilance and dedication of nurse anesthetists can lead the efforts to prevent adverse outcomes from surgical fires. PMID- 15354921 TI - Surgical management of testicular cancer. AB - Surgery is an integral component in the management of testicular cancer. Prior to the advent of cisplatin chemotherapy, a retroperitoneal lymph node dissection (RPLND) was the only chance for cure of testicular cancer. Over the years, the surgical techniques have been improved greatly to decrease the occurrence of complications (e.g., incidence of retrograde ejaculation). Currently, RPLND can be done as the initial therapy or after chemotherapy. In either situation, the postoperative management of patients with testicular cancer can be complicated and requires thorough, ongoing assessment. This article presents the surgical indications for RPLND and the nursing management. PMID- 15354922 TI - Prostate cancer recurrence fear: the prostate-specific antigen bounce. PMID- 15354923 TI - The challenges of long-term treatment outcomes in adults with malignant gliomas. AB - Malignant gliomas are among the most devastating tumors, with survival only one to three years after diagnosis even with the best of treatments. For this reason, long-term follow-up has not been established. Patients who do survive have significant disabilities, which compromise patient and family quality of life (QOL). Side effects from a brain tumor are dependent on the location of the tumor in the brain and can cause significant cognitive, emotional, and social effects on patients and families. Surgical treatment options also are dependent on the location of the tumor in the brain, and some sites remain inoperable (e.g., brain stem, motor strip). Radiation therapy continues to remain the mainstay of treatment for this patient population. The advent of newer technologies in radiation and chemotherapy and the development of novel treatments have increased survival. As survival increases, QOL issues become more important to patients and families. Impairment, disability, and handicap are measurements of QOL used by healthcare professionals. A multitude of studies has produced evidence regarding the impact of tumor location and size on QOL, whereas other studies have demonstrated the effects that various treatment modalities have on QOL. Few studies actually have provided insight into the everyday consequences that tumors pose on patients. Understanding the significance and causes of these impairments, disabilities, and handicaps will assist oncology nurses in caring for this special population. PMID- 15354924 TI - Management of patients receiving antithymocyte globulin for aplastic anemia and myelodysplastic syndrome. AB - Antithymocyte globulin (ATG) is used commonly in patients with severe aplastic anemia and those undergoing renal transplant. Its utility also is being explored in the treatment of myelodysplastic syndrome, conditioning regimens for hematopoietic stem cell transplant, and prophylaxis of graft-versus-host disease. As indications for ATG expand, knowledge regarding its administration and management of associated toxicities is needed. These toxicities range from life threatening anaphylaxis associated with the infusion to flu-like symptoms that occur one to two weeks after the infusion. Adverse effects are classified according to the severity and system impacted. Mild toxicities respond to comfort measures and include fever, chills, urticarial rash, and vomiting. Moderate toxicities require acute interventions and include fluid-responsive hypotension, nonischemic chest pain, and reversible oxygen desaturation. Severe toxicities require intensive support and include those refractory to earlier intervention. Management of these toxicities usually is limited to fluid resuscitation and noninvasive monitoring. Occurrence of infusion-related toxicities may require premature discontinuation of therapy. Therefore, an educated healthcare team and interdisciplinary clinical management guidelines are important to ensure the safe administration and complete course of ATG. PMID- 15354925 TI - New directions in oncology nursing care: focus on gefitinib in patients with lung cancer. AB - Treatment of solid tumors with chemotherapy regimens commonly is associated with debilitating or life-threatening side effects. Careful patient management, appropriate and prompt management of side effects, and interruption of therapy frequently are required for patients receiving chemotherapy. Furthermore, the systemic toxicity associated with chemotherapy may result in irreversible and incapacitating side effects, such as peripheral neuropathy, that lead to poor quality of life in patients. Gefitinib (Iressa, ZD1839, AstraZeneca Pharmaceuticals LP, Wilmington, DE) is a biologically based, molecular targeted therapy with a novel mechanism of action: selective inhibition of the epidermal growth factor receptortyrosine kinase (EGFR-TK) activity. Once-daily oral treatment with gefitinib is well tolerated. In clinical trials, treatment with gefitinib resulted in durable tumor responses and improvement in lung cancer related symptoms in patients with advanced non-small cell lung cancer who had received prior chemotherapy. Trials are under way to explore the full potential of gefitinib and additional EGFR-TK inhibitors for other solid tumors and in other treatment settings, including prevention. Biologically based, molecular targeted therapies such as gefitinib are providing new treatment options for patients and adding a new dimension to clinical practice for oncology nurses. PMID- 15354926 TI - Current treatment for hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. In the United States, the incidence of HCC has increased by nearly 75% since the 1980s. The rise in HCC diagnoses in the United States has been attributed to an increased number of patients infected with viral hepatitis and better diagnostic techniques. The management of HCC begins with diagnostic confirmation, followed by accurate staging. Historically, the prognosis for patients with HCC has been poor; however, improved surveillance and radiologic imaging techniques have led to earlier detection of HCC and an increased opportunity to treat patients. Treatment options for HCC include surgical and nonsurgical modalities. Surgical therapy, by way of partial hepatectomy or orthotopic liver transplantation, is the only potentially curative treatment for HCC, but most patients are not eligible for these procedures by the time of diagnosis. Palliative options include ablative techniques, radiation, and systemic therapies. As the incidence of this malignancy continues to rise, oncology nurses, who are an integral part of the multidisciplinary team caring for these patients, must be aware of current management for HCC. This article will provide an overview of the complex management of patients with HCC in the United States. PMID- 15354927 TI - Multiple myeloma: an overview. AB - Multiple myeloma (MM) is a malignant hematologic disorder involving plasma cells. In MM, immunoglobulin is overproduced, and patients can develop skeletal, hematologic, renal, and/or neurologic complications. The exact etiology of MM is unknown. The treatment for MM includes chemotherapy, antiangiogenic medications, and, most recently, a proteasome inhibitor. Nursing care for patients with MM requires close monitoring for infections and anemia, pain control, and education about the disease and treatment options. Further understanding of the pathophysiology of MM may lead to newer treatment options for the disease. PMID- 15354928 TI - Managing tissue necrosis associated with oral carcinoma. PMID- 15354929 TI - Tumor lysis syndrome. PMID- 15354930 TI - Alteplase (Cathflo Activase). PMID- 15354931 TI - Transporting patients receiving continuous infusion chemotherapy. PMID- 15354932 TI - Chemoprevention of breast cancer. PMID- 15354933 TI - Chemotherapy safety. PMID- 15354934 TI - [Analysis of point mutations in the ygeD, gyrA and parC genes in fluoroquinolones resistant clinical isolates of Chlamydia trachomatis]. AB - Resistance of 14 clinical isolates of C. trachomatis to fluoroquinolones, i.e. of ciprofloxacin, pefloxaxin and ofloxacin, was assayed. Three isolates with a high resistance degree to all 3 drugs (MIC equal or above 64 microg/ml) were detected. MIC was found to be equal to or below 4 microg/ml for 3 isolates. The remaining isolates had an intermediate resistance level. The nucleotide sequence was established for the Quinolone-Resistance Determining Region (QRDR) genes coding the DNA-gyrase subunit A (gyrA) and DNA-topoisomerase IV subunit C (parC) as well as for the 3'-region of ygeD coding, presumably, the efflux protein. In none of the isolates, the gyrA and gyrC QRDR differed from the corresponding regions in the published C. trachomatis genome sequence. Several silent mutations and mutations resulting in amino acid substitutions were observed in the ygeD 3' region of 2 isolates resistant to high FQ concentrations and in 1 isolate with the intermediate resistance level. PMID- 15354935 TI - [KatG Ser3 15Thr mutation as the main reason of isoniazid resistance in Mycobacterium tuberculosis isolated in the Novosibirsk and Kemerovo Regions]. AB - KatG Ser3 15Thr mutation is one of the main reasons of resistance to isoniazid in Mycobacterium tuberculosis isolates. The frequency rate of the above mutation among isoniazid-resistant isolates made 94% in Novosibirsk Region and 93% in Kemerovo Region. The use of PCR-restriction fragment length polymorphism (PCR RFLP) can be regarded as an adequate method for rapid screening of isoniazid resistance in Mycobacterium tuberculosis isolated in the West-Siberian Region. PMID- 15354936 TI - [PCR with subsequent sequencing of the 16S gene rRNA in species identification of mycobacteria of the non-tuberculosis complex]. AB - One hundred of mycobacterium cultures were assayed by the method of PCR with subsequent sequencing of the 16S rRNA region. The below mycobacterium species were identified: M. tuberculosis complex (n = 55), M. avium (n = 17), M. intracellulare (n = 4), M. scrofaleceum (n = 2), M. kansasii - M. gastri (n = 3), M. gordonae (n = 3), M. ulcerans - M. marinum (n = 1), M. smegmatis (m = 2), M. fortuitum (n = 11), M. peregrinum (n = 1) and M. chelonae - M. abscessus (n = 1). The method enabled the differentiation of species M. avium from M. intracellulare and M. peregrinum from M. fortuitum, which could not be differentiated by using the classic biochemical and bacteriological methods. Genetic heterogeneity of the mycobacterium strains of M. avium, M. fortuitum and M. gordonae was also established by PCR plus sequencing of the 16S rRNA region. PMID- 15354937 TI - [Monoclonal antibodies to B.anthracis protective antigen are capable to neutralize and to enhance the anthrax lethal toxin action in vitro]. AB - Anthrax belongs to highly dangerous infections of man and animals. No effective treatment methods for pulmonary types of the disease have been yet developed. The existing anthrax vaccines were designed decades ago and need improvement to fit the large-scale vaccination of population. At the same time, the immunological properties of the anthrax vaccine main component, i.e. of the protective agent, have been poorly studied. We obtained, within the present case study, a panel of mouse monoclonal antibodies to the protective agent and investigated the properties of the highest-affine panel representatives. An unusual phenomenon was detected, which is related with enhancement of the anthrax toxin action on the mouse macrophage-like cell-line in presence of the 1F2 monoclonal antibody. The remaining analyzed antibodies, i.e. 6G8 and 6G7, were found to neutralize effectively the toxin action. The enhancing and neutralizing antibodies were proven to be specific to different domains of the protective antigen and to recognize epitopes in its composition. The antibody-mediated enhancement of the anthrax lethal action is a convincing argument for further development of a new generation anthrax vaccine. Definition of the linear antigen determinants for neutralizing antibodies in the protective antigens is an important step in the development of the next-generation anthrax vaccine. PMID- 15354938 TI - [Interaction between the human and simian adenoviruses in human cells: complementation, transcapsidation and formation of defective adeno-adeno hybrids]. AB - It was for the first time that complementation between the human and simian adenoviruses in human cells as well as the ability of the human adenovirus Ad2 (HADv2) genome to transform completely into the simian adenovirus SA7(C8) (SADv15) capsid (transcapsidation) was demonstrated. A defective adeno-adeno hybrid (recombinant) between the above viruses is described; the recombinant has the SA7(C8) capsid and Ad2 genome with a 10% insertion of SA7(C8) in the central region. Defective hybrid virions are able to replicate both in human and simian cells by using the SA7(C8) virus as helper. The hybrid virions help the above virus to replicate in human cells: they form a mutually complementing virion pair. PMID- 15354940 TI - [Single insertions of omegon into Azospirillum brasilense DNA result in defects in the bacterial mobility of several types]. PMID- 15354941 TI - What is emergency care? PMID- 15354942 TI - A two-year clinical study of metal-ceramic and metal-polymer crowns. AB - The purpose of this study was to compare full metal crowns with two different types of veneering material - ceramics and polymer after a two-year period in a function. The aim was to evaluate the changes of occlusal relief, marginal adaptation, break of material, changes of shape and color and gingival status. One hundred and two crowns were examined in 34 patients immediately after cementation, and they were reviewed within two years of crown placement. These fixed restorations were checked according two modified US Public Health Service System criteria. After two years the crowns were found clinically intact in 94% for metal-ceramics versus 98% for metal-polymer. No crown was lost. There were found significant differences in positions of a crown margin between both materials and marginal adaptation between initial (baseline) and follow-up results. Gingival health at baseline was related in optimal in 67% versus 77%, after two years in 43% versus 71%. Slight mismatch in color occurred in 0 versus 60% crowns, obvious mismatch was apparent in 0 versus 21% crowns after two years. Within the limitations of this study, it was found that the examined metal crowns with two different veneerings are very stable fixed restorations, mechanical resistance and integrity of polymer material was surprisingly good but its color stability presented mismatch after two-years period. Ceramic veneering was excellent in color match but there were three visible cracks of the ceramic layer. PMID- 15354943 TI - Oxidative stress, inflammation and autoimmune reaction in type 1 and type 2 diabetes mellitus. AB - Diabetes mellitus (DM) is associated with oxidative stress, elevation of inflammatory markers and other mechanisms, which may contribute to accelerated atherosclerosis. The aim of the study was to determine prominent factors of these pathogenic processes in patients with DM, to examine their relationship in serum, and to find out the differences between DM1 and DM2. Advanced oxidation protein products (AOPP), C-reactive protein (CRP), pregnancy-associated plasma protein-A (PAPP-A), anticardiolipin antibodies (ACA) and anti-beta2-glycoprotein-1 antibodies (anti-beta2-GPI) were determined in 27 patients with DM1, 27 patients with DM2 and 23 healthy subjects. AOPP, CRP and anti-beta2-GPI were significantly elevated in DM2 in comparison with healthy subjects (p<0.01, p<0.0001, p<0.0001, respectively). In DM1, anti-beta2-GPI were elevated (p<0.0001) as well, but there was no increase of either AOPP or CRP. There was no difference in PAPP-A levels in DM1 or DM2 and healthy subjects. In DM 1, AOPP correlate significantly with anti-beta2-GPI (r = 0.68, p<0.05). In DM2, there is a significant correlation between anti-beta2-GPI and PAPP-A (r=0.45, p<0.05). Oxidative stress and inflammation are more expressed in DM2 and they are partly related. In DM1, oxidative stress seems to be in closer link to autoimmune reaction than to inflammation. PMID- 15354944 TI - The examination of the temporomandibular joint on 1,5T magnetic resonance. AB - The most frequent intraarticular disorder of the temporomandibular joint (TMJ) represents the disorder of the functional relation between the articular disc and mandible condyle, i.e. dislocation of the articular disc. Magnetic resonance (MR) optimally visualizes the soft and hard articular tissues (articular disc, articular socket and condyle of the mandible) and its surroundings. The aim of the study was to evaluate findings of MR investigation of the TMJ. We investigated billateraly 26 patients with TMJ disorders during the years 1996 and 2003. All patients were examined on 1,5 T MR. Joints were studied in T1 and T2 weighted images with closed mouth and during mouth opening in dynamic study. MR verified dysfunction was observed in 48 investigated TMJ (92,3 %), hydrops of the joint was observed in 3 joints (5,8 %), arthrosis of the condylar head in 4 patients (7,6 %). Only four TMJ had normal MR finding (7,6 %). MR represents the best method for studying clinically affected joints, for the evaluation of the morphological state of TMJ and the analysis of the dynamic process during mouth opening. Method is also useful for revealing of a disorder in clinically silent joints. T2 weighted image in TSE mode brought best imaging of the joint. The fat saturation sequence was advantageous mainly in liquid storage (hydrops of the joint, edema in the adjacent bone). The dimensions of the articular cleft and bone components of the joint were well-visualized in T1 weighted images. PMID- 15354945 TI - Agenesis of third molars in young Czech population. AB - The aim of this study is to evaluate the incidence of third molars agenesis in the Czech population. One thousand patient's panoramic radiographs of the pedostomatological department of Second Stomatological Clinic of The First Faculty of Medicine, Charles University, has been analyzed. The age of probands was ranging between 12 to 21 years inclusive. Patients were divided into 10 groups according to age. In each age group were 100 individuals - 50 boys and 50 girls. Agenesis of 1-4 third molars was present in 22.5% (25.6% in boys and 19.4% in girls). Intersexual difference is significant. Further, the incidence of third molars agenesis in individual dentition quadrants, upper and lower jaw and right and left side of dentition was examined. Neither in boys nor in girls were detected significant differences in these values. Agenesis of just one, any third molar occurs in 10.9% (12.0% in boys and 9.9% in girls). For one individual it is mean 1.9 congenitally missing third molars in boys and 2.0 congenitally missing third molars in girls. These intersexual differences are not significant. In boys, the most frequent agenesis was of one, next of two, then of three and at last of four third molars. In girls, agenesis of two was on the first place, then that of one and lastly and equally of three and of four third molars. The results were compared with literature data. PMID- 15354946 TI - QT dispersion estimated from 80 body surface potential map leads and from standard 12-leads ECG in psychiatric patients treated with dosulepin. AB - The aim of the study was to detect changes of the QT dispersion (QTd) due to cardiotoxicity of tricyclic antidepressant dosulepin. Electrocardiographic and body surface potential mapping (BSPM) recordings were obtained using Cardiag 112.2 diagnostic system from 27 psychiatric outpatients treated with prophylactic doses of dosulepin and compared to those obtained from 37 healthy volunteers. From these recordings the QTd and the dispersion of heart rate-corrected QT interval QTc were evaluated. These parameters were estimated both from 80 BSPM leads and from 12 standard ECG leads. Acquired data were statistically correlated by Spearman rank order correlation coefficient with dosulepin plasma levels. The average QTd evaluated from BSPM leads (+/-SD) in the dosulepin group was significantly higher [70 (+/-21) ms] than that in the control group [34 (+/-12) ms] (P< 0.001). Moreover, the correlation between QTd and the dosulepin plasma level was statistically significant as well (P< 0.001) with the value of correlation coefficient 0.7871. The QTd evaluated from standard 12 ECG leads was increased in dosulepin group as well [46 (+/-18) ms vs. 28 (+/-10) ms - P< 0.05] but we have not found any significant correlation of the QTd with the dosulepin plasma level. According to the above-mentioned results we can conclude that the QTd estimated from BSPM leads (but not that estimated from 12-lead ECG) could be used as a marker of the dosulepin effect on the myocardium. PMID- 15354947 TI - Daily oral versus pulse intravenous cyclophosphamide in the therapy of ANCA associated vasculitis--preliminary single center experience. AB - The aim of the multicentric randomized trial CYCLOPS is to optimize the treatment of induction of remission in patients with generalized, but not immediately life threatening ANCA (antineutrophil cytoplasmic antibodies) -associated vasculitis. This will be achieved by reducing the dose of cyclophosphamide by administering it as intermittent pulses. The lower cumulative dose will be very probably accompanied with lower toxicity, whereas the effectivity should be comparable. We have enrolled 28 patients to the study. At present, 18 of them are suitable for evaluation. Our preliminary results show that pulse intermittent administration of cyclophosphamide is safer from the point of morbidity and mortality due to infectious complications. In our hands, this treatment modality does not seem to be less effective than the conventional daily oral cyclophosphamide. However, unambiguous results and treatment recommendations will not be available until the final evaluation of all patients enrolled in the trial. PMID- 15354948 TI - Focal segmental glomerulosclerosis in solitary kidney in WAGR syndrome. AB - WAGR syndrome consists of Wilms' tumour, aniridia, genitourinary malformations and mental retardation, and is associated with chromosomal microdeletion of 11p13. We report a case of young male, exhibiting several typical features of WAGR syndrome (e.g. WT, aniridia and genitourinary abnormalities), but missing some other (mental retardation and chromosomal abnormality absent). Renal biopsy performed in our patient for unexplained proteinuria showed focal segmental glomerulosclerosis, presumably of secondary origin; the decrease of proteinuria was achieved by the firm control of BP in conjunction with the reduction of body weight. PMID- 15354949 TI - One hundred semesters of educational and scientific work accomplished for Charles University in Prague and its First Faculty of Medicine by Professor RND. Vladimir Novotny, DSc. PMID- 15354950 TI - [Pattern of skeletal muscle differentiation in fish: molecular and biological approaches]. AB - The initial stages of myogenesis going in myoblasts include the stages of induction, determination, and differentiation. The induction and determination of cells in the myotomes are controlled by morphogenetic signals from neighboring tissues of the notochord and neural tube manifested as expression of genes of Shh and Wnt families, respectively. In fish (at the example of danio), this signal is passed to somite cells neighboring the notochord; later the cells migrate to the embryo surface and differentiate into slow muscle fibers. Synthesis of the main contractile proteins, primarily the components of myosin molecule--heavy chain (MHC) and individual isoforms of light chains (MLC1, MLC2, and MLC3)--are encoded by different genes during different ontogenetic stages. The peptide maps obtained after alpha-chymotrypsin digestion of MHCs from larvae, fast and slow skeletal muscle of loach are different, which points to differences in their primary structure. In addition, considerable differences were revealed in the structure of MLC isoforms at different ontogenetic stages. The definitive fast muscle contained three light chain types, MLC1, MLC2, and MLC3; slow muscle, MLC1 and MLC3; while the larval muscle fibers included a specific larval MLCL in addition to MLC3. PMID- 15354951 TI - [Oxides of nitrogen (NO* and NO2-) as cofactors of the myeloperoxidase system]. AB - Myeloperoxidase is the main peroxisomal protein of neutrophils, monocytes, and a subpopulation of tissue macrophages; it plays the key role in protective and inflammatory responses of the organism. This role is mediated by various diffusible radicals formed during oxidative reactions catalyzed by the enzyme heme. Myeloperoxidase and nitric oxide synthase are stored in peroxisomes. Nitric oxide reacts with the heme of myeloperoxidase. Low nitric oxide concentrations increase peroxidase activity through reduction of Compound II to native myeloperoxidase. Conversely, high nitric oxide concentrations inhibit the catalytic activity of myeloperoxidase through formation of inactive nitrosyl-heme complexes. Such effect of nitric oxide on catalytic activity of myeloperoxidase has various consequences for infectious and local inflammatory processes. Another oxide of nitrogen, nitrite, is a good substrate for myeloperoxidase Compound I but slowly reacts with Compound II. Nitrogen dioxide is formed after nitrite oxidation by myeloperoxidase. Formation of nitrogen dioxide is another protective mechanism and nitration of microbial proteins by myeloperoxidase can represent an additional protective response of peroxisomes. PMID- 15354952 TI - [Apoptosis in P388 leukemia cells induced by specific inhibitors of 5- and 12 lipoxygenase and the product of cyclooxygenase, prostaglandin E2]. AB - We studied the effect of specific inhibitors of 5- and 12-lipoxygenases as well as the product of cyclooxygenase activity, prostaglandin E2, on proliferation and death of P388 leukemia cells. Inhibition of 5- and 12-lipoxygenases in the cells inhibits proliferation and induces apoptosis. The concentrations of baicalein, an inhibitor of 12-lipoxygenase, and AA861, an inhibitor of 5-lipoxygenase, causing a 50% death rate (LC50) proved to be the same, 50 microM. Excessive prostaglandin also inhibited proliferation of the cells and induced apoptosis. The LC50 for prostaglandin E2 was 4 microM. The obtained data suggest that apoptosis in P388 cells after lipoxygenase inhibition can be induced by both deficiency of lipoxygenase products and excess of prostaglandins in the cell. PMID- 15354953 TI - [Regenerative capacity of muscle tissue and thymus state in irradiated rats exposed to long-term He-Ne laser radiation]. AB - We studied the effect of He-Ne laser on regeneration of damaged gastrocnemius muscle in rats irradiated at 6 Gy in conditions of fractional laser energy spread (10 exposures, 3 min for each limb, within 30 days after the operation; 2-3 exposures weekly; 2.5-3.0 mW/cm2 power density; and 9.0-10.8 J/cm2 total dose per animal). Laser radiation stimulated regenerative activity of the skeletal muscle and favored a more even distribution of load on the thymus (a smooth decrease in its weight and slow aplasia). The level of chromosomal aberrations in the thymocytes demonstrated certain instability although remained lower as compared to the control during the whole observation period (60 days). PMID- 15354954 TI - [Effect of vernalization and red light illumination of seedlings of bread wheat (Triticum aestivum L.) on the temperature profile of the cAMP phosphodiesterase activity]. AB - Phenotypic manifestations of Vrn (vernalization) and Ppd (photoperiodism) genes responsible for transition of bread wheat Triticum aestivum L. to generative growth (flowering) are mutually related. Since the mechanism of phytochrome induced photoperiodism involves the enzymes of cyclic adenosine monophosphate metabolism and phosphodiesterase in particular, we tested involvement of phosphodiesterase in the process of winter wheat vernalization and formation of flowering competence in alternate wheat requiring a long day but no vernalization for the transition to flowering. We studied temperature dependence of phosphodiesterase activity in vernalized and unvernalized winter wheat on the one hand and in etiolated and red light illuminated seedlings of alternate wheat on the other hand. Short-term experiments demonstrated that vernalization and red light illumination are similar to long day by the effect on the long-day plants. Both influences induced a pronounced inversion of the temperature profile of phosphodiesterase activity in the 28-45 degrees C range. We propose that phosphodiesterase is involved in vernalization processes and can serve as a sensor of low temperature in winter wheat. Changed temperature profile is a radical control mechanism of phosphodiesterase activity in response to the influences (red light and vernalizing temperatures) responsible for competence of various bread wheat forms for generative growth. PMID- 15354955 TI - [Physicochemical and immune properties of glycoglycerolipids from Laminaria japonica within immunostimulating complexes (ISCOMs)]. AB - Certain physicochemical properties of glycoglycerolipids from marine alga Laminaria japonica (monogalactosyl diacylglycerol, digalactosyl diacylglycerol, and sulfoquinovosyl diacylglycerol) and their ability to be incorporated into immunostimulating complexes (ISCOMs) used for presentation of microbial and tumor antigens in vesicular form were comparatively described. These glycolipids proved to considerably differ by fatty acid composition, degree of unsaturation, and phase transition temperature. Possible production of modified ISCOMs through incorporation of these glycolipids into the vesicle instead of the glycolipid component was demonstrated. Preliminary data demonstrated no significant increase in immune response to Yersinia pseudotuberculosis porin within the modified (with monogalactosyl diacylglycerol) and classical (with phosphatidylcholine) ISCOMs as compared to individual porin. PMID- 15354956 TI - [Combined effect of diazepam and GABAA-ergic ligands on the activity of Cl(-) ATPase from plasma membrane of bream brain (Abramis brama L.)]. AB - We studied the combined effect of diazepam and GABAA-ergic ligands on the activity of Cl- -ATPase from plasma membrane of bream brain. of The membranes were preincubated and incubated with diazepam as well as with other GABAA-ergic ligands at physiological pH (7.4), i.e. under the conditions when Cl- -ATPase activity is undetectable. GABA (0.1-100 microM) induced Cl- -ATPase activity with the maximum effect at 10 microM. Diazepam (0.1 microM) enhanced the effect of low GABA concentrations (0.1-1 microM) on Cl- -ATPase activity but had no effect on the enzyme in the presence of high GABA concentrations (10-100 microM). At the same time, GABA (1 microM) enhanced the effect of low diazepam concentrations (0.1-1 microM) on the enzyme activity but had no effect on it in the presence of high concentrations of the ligand. Blockers of GABAA-ergic receptors, picrotoxin (50 microM) and bicuculline (5 microM), canceled the combined effect of diazepam and GABA on the enzyme activity. The obtained data demonstrate that the combined effect of diazepam and GABAA-ergic ligands on Cl- -ATPase activity at physiological pH is similar to the effect of these ligands on GABAA/benzodiazepine/Cl- channel. PMID- 15354957 TI - [Genetically transformed plant roots as a model for studying specific metabolism and symbiotic contacts of the root system]. AB - Genetic transformation of plants mediated by Ri plasmid of Agrobacterium rhizogenes occupies a special place in plant cell engineering, since this technique based on a natural phenomenon allows cultivation of separately growing roots on hormone-free media. Application of wild-type unmodified agrobacterial strains allows us to obtain root cultures capable of long-term growth in vitro due to an increased sensitivity of the cells to auxins while other biochemical properties remain unaltered. A collection of pRi T-DNA transformed roots of certain dicotyledons was constructed; some strains of it are used to study synthesis of secondary metabolites in root cells. The in vitro cultivated roots could synthesize root-specific metabolites, which allows their large-scale application for biotechnological production of ecologically pure crude drugs. Cocultivation of pRi T-DNA transformed roots with arbuscular mycorrhizal fungi makes possible vital study of all stages of obligate symbiont development and interaction with plant roots. Mixed axenic culture of AM fungi and pRi T-DNA transformed plants can be used to construct a collection of the most valuable endomycorrhizal fungal species and to produce considerable quantities of homogeneous fungal inoculums. PMID- 15354958 TI - [Spatial distribution of lectin activity in perilesional areas of tobacco leaves inoculated with tobacco mosaic virus]. AB - The activities of lectins and peroxidase and lignin content were studied in the perilesional area of leaves in two tobacco species (Nicotiana tabacum L., cultivar Samsun NN, and N. glutinosa L.) inoculated with tobacco mosaic virus. The development of hypersensitivity response proved to be accompanied by a complex spatial and temporal distribution of lectin activity. The area 5-9 mm away from the lesion center was characterized by the highest activity of loosely bound membrane lectins eluted with 0.05% Triton X-100. In the fraction of tightly bound membrane lectins (eluted with 0.5% detergent), lectin activity decreased during the first two days but increased on day 4 after inoculation. The activity of loosely bound membrane lectins increased in the leaf areas distant from the lesion. Two-phase dynamics in the interlesional area were also observed for lectin activity in the tightly bound membrane fraction (decrease on day 2 days and increase on day 4 after inoculation) and for peroxidase activity (increase on day 2 days and decrease on day 4). The relationship between the dynamics and spatial distribution of lectins in the perilesional area and the possible involvement of these proteins in pathogen-induced changes in photosynthesis are discussed. PMID- 15354959 TI - [Photosystem damage and spatial architecture of thylakoids in chloroplasts of pea chlorophyll mutants]. AB - The effect of chlorophyll-protein complexes on the ultrastructure of chloroplasts was studied in the leaves of pea, the parent cultivar Torsdag and mutants chlorotica 2004 and 2014. The mutants were shown to accumulate 80 and 55% of chlorophyll, relative to the control, while the composition of the synthesized photosystem complexes was the same as in the parent cultivar Torsdag. The size of the light-harvesting antenna was similar to the control in the 2014 mutant but considerably increased (by 30%) in the 2004 mutant. These changes were due to a proportional decrease in the number of all complexes (by 40-45%) in the 2014 mutant. At the same time, the number of reaction center complexes of photosystem I (PS I) decreased by 50% while that of photosystem II (PS II) remained virtually constant in the 2004 mutant. A proportional decrease in the number of the PS I and PS II complexes in the chlorotica 2014 mutant was accompanied by a partial reduction of the entire chloroplast membrane system against the background of normal development of both granal and intergranal sites of thylakoids. Conversely, the loss of PS I reaction centers led mainly to the reduction of the intergranal sites of thylakoids in chloroplasts. This effect is attributed to the prevalence of PS I complexes in the intergranal thylakoids. PMID- 15354960 TI - [Mechanisms of responses of systemic circulation: role of endothelial factor in the vascular tone regulation]. AB - Experimental data on the effect of NO synthase inhibition on hemodynamic changes (blood pressure, cardiac output, and peripheral resistance) induced by an increased (polyglucin infusion) or decreased (orthostasis) cardiac output are presented. Under conditions of NO synthase inhibition, the pressor effects of polyglucin and orthostatic hypotension increased by 70 and 72%, respectively. The response of peripheral resistance had a similar trend. Significance of NO secretion by vascular endothelium for the development of systemic hemodynamic responses is proposed. PMID- 15354961 TI - [Antidiabetic and anticoagulant properties of the heparin-glutamic acid complex]. AB - Natural heparin complexes proved to activate the anticoagulation system. The obtained experimental data convincingly confirm that glutamic acid alone, and particularly in a complex with heparin, has a considerable preventive potential and efficiently protects experimental animals with induced diabetes mellitus. PMID- 15354962 TI - [Ecological and biochemical evaluation of the environment of modern foraminifers of White Sea]. AB - Current ecological conditions in the regions of Bol'shoi Keretskii raid and Glubokaya Salma (Kandalaksha Bay, White Sea) were evaluated on the basis of structural and dynamic indices of the upper sediments in the context of specific composition of the benthic foraminiferal community. The distribution of the main species of living foraminifers was determined for the shallow and deep-sea groups. The relationship between populations of the most abundant species of living benthic foraminifers and hydrolytic enzyme activities providing for destruction of bottom organic matter at biochemical rates has been revealed. PMID- 15354963 TI - [Rank distributions in community ecology from the statistical viewpoint]. AB - Traditional statistical methods for definition of empirical functions of abundance distribution (population, biomass, production, etc.) of species in a community are applicable for processing of multivariate data contained in the above quantitative indices of the communities. In particular, evaluation of moments of distribution suffices for convolution of the data contained in a list of species and their abundance. At the same time, the species should be ranked in the list in ascending rather than descending population and the distribution models should be analyzed on the basis of the data on abundant species only. PMID- 15354964 TI - [Resistance to drying in aquatic mollusks]. AB - Morphological, behavioral, physiological, and biochemical adaptations of marine and freshwater mollusks favoring their survival in conditions of drying were reviewed. PMID- 15354965 TI - [Changes in ecological features of soils after controlled fires in forests defoliated by the Siberian moth in the southern taiga subzone of the Enisei Region, Siberia]. AB - Data on the postfire dynamics of soil properties in the foci of Siberian moth population outbreaks are considered. It has been shown that controlled fires set in pest-defoliated forests result in the loss of approximately 75% of carbon and 50% of nitrogen from the forest litter through their emission into the atmosphere and in the enrichment of the upper soil horizons with potassium and phosphorus (this concerns both total and movable forms). Microbiological processes in the organogenic horizon undergo significant transformation, the density of microarthropods decreases, and the abundance of mites becomes hundreds of times lower. PMID- 15354966 TI - End-stage renal care in developing countries: the India experience. AB - Chronic renal failure is a devastating medical, social and economic problem for patients and their families. There is no data on the true incidence and prevalence of chronic renal failure in the developing world. Delayed diagnosis and failure of institution of measures to slow progression of renal failure result in a predominantly young ESRD population. Renal replacement therapy (RRT) is a low-priority area for healthcare planners in developing nations with two tier healthcare delivery system. There is a severe shortage of nephrologists and hospitals offering dialysis and transplantation, more so in the poorest regions. There is a direct relationship between the number of dialysis centers and per capita gross national income of developing nations. Shortage in the number of government-funded hospitals has fanned the growth of a large number of private hospitals offering RRT. The high cost of hemodialysis (HD) puts it beyond the reach of all but the very rich and maintenance HD is the exclusively preserve of private hospitals. Government-run hospitals are busy with renal transplantation, which is the only realistic long term RRT option for a majority of patients. There are no state-funded or private health insurance schemes and patients have to raise finances for RRT on their own. Entire families are involved in such endeavors, with resulting loss of income of other family members too. A number of measures are utilized to bring down the RRT costs. For HD, these include cutting down the frequency of dialysis, use of cheaper cellulosic dialyzers, dialyzer reuse and nonutilization of expensive drugs like erythropoietin. Paradoxically, chronic peritoneal dialysis is more expensive than HD; patients use outdated connection systems and are suboptimally dialyzed on 3 exchanges/day. Most patients on dialysis are inadequately rehabilitated. Renal transplant recipients are forced to discontinue expensive drugs like cyclosporine after variable periods leading to high rates of graft loss. Financial considerations often preclude appropriate treatment of steroid-resistant rejection and cytomegalovirus infection. There is no organized cadaver donation program and an overwhelming majority of transplants are performed using living donors. This led to the practice of the sale of kidneys for transplant. To conclude, the financial burden of RRT in developing nations impacts on the lifestyle and future of entire families, and extracts a cost far higher than the actual amount of money spent on treatment. PMID- 15354967 TI - Strategies for the treatment of acute renal failure in intensive care units: the aspect of dosing. AB - Despite all the medical progress, the mortality rate in intensive care units for patients with acute renal failure (ARF) remains high, among specific patient populations, up to 88% [Letourneau I, Dorval M, Belanger R, Legare M, Fortier L, Leblanc M. Acute renal failure in bone marrow transplant patients admitted to the intensive care unit. Nephron Apr 2002; 90 (4), 408-412.]. Recent trial results indicate that patient survival may be improved by adequate renal replacement therapy. In particular, the dose of intermittent and continuous renal replacement therapies has proved to be a significant factor affecting patient survival. Daily intermittent hemodialysis, e.g., is superior to alternate-day intermittent hemodialysis, and with continuous therapies, survival is related to the filtration rate. Further relevant factors include early initiation of renal replacement therapy, choice of biocompatible membranes and the application of bicarbonate-buffered replacement solutions for defined patient groups. The advantages offered by continuous techniques could be demonstrated for individual patient groups; in meta-analyses, advantages were shown for the total population of patients with ARF. Other than for patients with chronic renal failure (NKF DOQI. Clinical practice guidelines for hemodialysis adequacy. Am J Kid Dis 1997; Vol. 30, 515-566.), there are no current clinical guidelines for a standard treatment of intensive care patients with ARF. Therefore, such a treatment standard still needs to be determined. PMID- 15354968 TI - Acute renal failure during eclampsia: incidence risks factors and outcome in intensive care unit. AB - OBJECTIVE: To assess the incidence, risk factors and the outcome of acute renal failure (ARF) associated with eclampsia in intensive care unit (ICU). DESIGN: Prospective and analytic study. SETTING: A surgical ICU in a university hospital. PATIENTS: 178 consecutive women with eclampsia admitted to an intensive care unit during seven years. ARF was defined by a serum creatinine concentration >140 micromol/L. RESULTS: The incidence of ARF was 25.8%. In univariate analysis the severity of patient illness, the complications associated with eclampsia (disseminated intravascular coagulation, Hellp syndrome, neurologic complications, abruptio placenta, aspiration pneumonia, delivery hemorrhage) were significantly associated with ARF. In a logistic regression model, risk factors for ARF included organ system failure (OSF) odds ratio (OR)=1.81 confidence interval (CI) [1.08-3.05], bilirubin >12 micromol/L OR=4.42 CI [1.54-12.68], uric acid >5.9 g/dL OR=16.5 CI [3.09-87.94], abruptio placenta OR=0.2 7 CI [0.08 0.99], and oliguria OR=0.10 CI [0.03-0.44]. In contrast, severity of blood pressure or proteinuria on dipstick were not associated with ARF. However, in this series, 15 women required dialysis in the short term and one required long term dialysis. ARF associated with eclampsia was significantly associated with mortality (32.6% versus 9.1% p=0.0001). CONCLUSION: ARF with eclampsia is a frequent situation that required intensive management when risks factors were present. The need for dialysis was a rare condition. PMID- 15354969 TI - The role of intrinsic fibrinolytic system activation in pathogenesis of hemostasis disturbances in hemodialyzed patients with chronic renal failure. AB - In the hemodialysis patient, hemostasis changes may occur. The contribution of fibrinolysis in pathogenesis of these disorders is unclear. The aim of the study was to estimate intrinsic fibrinolysis pathway in patients treated with hemodialysis (HD) because of chronic renal failure caused by chronic glomerulonephritis. The study was performed with 43 patients; the control group consisted of 51 healthy volunteers chosen by sex and age. The following parameters were determined: concentration of the urokinase plasminogen activator antigen (uPA:Ag), plasmin--antiplasmin complexes (PAP), fibrin and fibrinogen degradation products (FDP), activity of prekallikrein (PK) and C1-inhibitor (C1 INH) and also euglobulin clot lysis time (ELT). The above parameters were assessed in the patients before and after HD and were compared with the control group. In the HD patients, in comparison with the control group, prolonged statistically ELT [153 (125;215) vs. 105 (75;142) min.; p<0.001], with increase of PAP (508.6 +/- 274.7 vs. 184.7 +/- 69.4 microg/L; p<0.001) and FDP concentrations [5 (5;15) vs. 2.5 (0;0.3) microg/mL; p<0.05] before the procedure were determined. It suggests increased plasmin production and fibrin digestion despite determination of decreased general fibrinolytic activity. The C1-INH activity before HD was also significantly increased as compared with the control group [157 (136;171) vs. 107 (100;124)%; p<0.001], and its significant decreased after the HD is 157.7 +/- 23.9 vs. 122.3 +/- 20.3%; p<0.001, as it seems to be a nondirect proof of intrinsic pathway contribution in fibrinolysis activation in the HD patients. The remaining examined parameters did not change significantly after the dialysis procedure. PMID- 15354970 TI - Hemodynamic maladjustment and disease progression in nephrosis with FSGS. AB - Idiopathic nephrotic syndrome (NS) associated with focal segmental glomerulosclerosis (FSGS) and severe renal function impairment is usually refractory to the conventional treatment and progresses to end-stage renal disease. Herein, we reported 10 patients with NS-FSGS who had initially had CCr 34 +/- 12 mL/min/1.73 m2 (normal 120 mL/min/1.73 m2), FE Mg 7.8 +/- 2.6% (normal 2.2%), 24-h urinary protein 3.1 g (normal <200 mg) and been followed up for over 10 years. The initial intrarenal hemodynamic study revealed a marked elevation of efferent arteriolar resistance (RE 17289 +/- 8636 dyne x s x cm(-5); normal 3000 dyne x s x cm(-5)), intraglomerular hypertension (PG 57 +/- 1 mm Hg; normal 52 mm Hg), hyperfiltration (FF 0.24; normal 0.2), marked reductions in GFR 35 +/- 17 mL/min/1.73 m2, renal plasma flow (RPF 159 +/- 61 mL/min/1.73 m2; normal 600 mL/min/1.73 m2) and peritubular capillary flow (PTCF 123 +/- 57 mL/min/1.73 m2; normal 480 mL/min/1.73 m2). Such a hemodynamic alteration indicated a hemodynamic maladjustment with a preferential constriction at RE. Treatment consists of multidrugs, namely angiotensin converting enzyme inhibitor, calcium channel blocker, antiplatelet and anticoagulant, with or without angiotensin II receptor antagonist. Following the treatment, correction of hemodynamic maladjustment has been achieved which is characterized by reductions in RE 6046 +/- 2191 dyne x s x cm(-5), PG 52 +/- mm Hg, FF 0.19 +/- 0.1 and increments in RPF 341 +/- 118 mL/min/1.73 m2, PTCF 280 +/- 106 mL/min/1.73 m2 and GFR 64 +/- 17 mL/min/1.73 m2. Coinciding with hemodynamic improvement, there has been a steadily increased creatinine clearance and improvement in FE Mg 4.3 +/- 2.6% and suppression of proteinuria 0.29 +/- 0.4 g/24 h after the period of follow-up of greater than 10 years. PMID- 15354971 TI - The evaluation of renal osteodystrophy with cortical quantitative ultrasound at various bone sites. AB - BACKGROUND: In this study we evaluated bone abnormalities of patients with chronic renal failure (CRF) by cortical quantitative bone ultrasound (QUS) measurements at different bone sites because these abnormalities may be of variable etiology and may start before symptoms or radiological changes are manifested. METHODS: Of fifteen patients with chronic renal failure, seven had moderate-severe disease and eight had been on chronic hemodialysis from 5 to 17 years, with renal osteodystrophy (ROD) confirmed by bone biopsies. Twelve normal subjects of similar age and gender volunteered for the control group. RESULTS: Patients and controls differed in creatinine clearance, in serum phosphate levels and in serum total alkaline phosphatase. Mean intact-PTH levels differed significantly amongst the three groups of subjects. All patients with ROD had intact-PTH higher than 200 pg/mL. The cortical ultrasound parameter, speed of sound (SOS), was slower in patients with more severe renal failure at all bone sites measured. The group with ROD had significantly lower cortical ultrasound values than the other patients and the control group at all sites. SOS values at the proximal phalanx, distal radius and midtibial sites were positively and significantly correlated. Cortical ultrasound measurements at the radial site correlated with midtibial and phalangeal sites but the correlation between midtibial and phalangeal sites did not reach significance. IPTH levels correlated negatively and significantly with cortical QUS values at all sites being the correlations higher at phalangeal and radial sites than at the midtibial region. CONCLUSIONS: The differences in cortical ultrasound observed indicate the potential clinical application of this methodology to evaluate bone abnormalities in chronic renal failure, especially in patients on chronic hemodialysis. PMID- 15354972 TI - Relationships between HLA-A, -B, -DQ and -DR antigens and interstitial fibrosis in renal allografts. AB - Although renal transplant recipients tend to exhibit similar clinical and immunological changes over time, some allograft biopsies show early IF. To evaluate the relationship between HLA antigens and IF in renal allografts, we reviewed for HLA-A, -B, -DQ, and -DR antigens in 88 renal transplant recipients. For each antigen type, we determined the numbers of patients who did and did not possess the antigen. We then determined the mean time to onset of IF for each of these two groups, and statistically compared the means. In the second part of the analysis, we divided the 88 patients into those who did and did not show IF at 6 months posttransplantation, and then calculated the prevalence of each antigen type in the IF (+) and IF (-) groups. This same procedure was repeated with the patients grouped according to presence of IF at 12 months posttransplantation. The patient groups with HLA-B8, -B27, -DQ2, -DQ5, -DQ6, -DQ7, -DR4, -DR13, and DR15, respectively, had significantly shorter times to IF onset after transplantation than the corresponding groups without these antigens (p<0.05). The groups that were IF (+) at 6 and 12 months posttransplantation had significantly higher frequencies of HLA-B8, -B27, -DQ2, and -DR4 than the corresponding IF (-) groups at these two time points (p<0.05). The results indicated that any kidney recipient with these antigens is predisposed to developing diffuse IF in their graft relatively soon after transplantation. We conclude that although there are multiple etiological agents in the pathogenesis of interstitial fibrosis, one cannot exclude an HLA association in these cases. PMID- 15354973 TI - Relationship of HLA-DR expression to rejection and mononuclear cell infiltration in renal allograft biopsies. AB - Research on renal biopsies has shown that HLA class I antigens are distributed throughout the renal parenchyma, but that the distribution of HLA-DR varies greatly. We investigated HLA-DR expression in biopsies of 90 renal transplants, and also semiquantitatively assessed the proportions of CD68-, CD3-, and HLA-DR positive infiltrating cells by immunohistochemistry. The relationships between tubular DR expression and interstitial lymphocyte and macrophage infiltration were examined. Forty of the biopsies showed acute rejection (AR), 33 showed chronic rejection (CR), 10 showed suspected rejection (SR), and 7 showed no evidence of rejection (NR). HLA-DR expression was noted in 35/40 (87.5%) of the AR cases, 22/33 (66.6%) of the CR cases, and 6/10 (60%) of the SR cases. Only 1 (14.3%) of the NR cases exhibited HLA-DR antigen expression in the renal tubules. The proportions of lymphocyte and macrophage infiltration observed in the interstitium were significantly correlated with tubular DR expression in all cases (p<0.01). At 6 months after biopsy was done, 24/35 (68.6%) of the AR patients with tubular DR expression had showed second episode of rejection or showed deteriorated renal function. The remaining 11 AR cases with tubular DR expression had stable renal function at this stage. The cases that had no significant tubular DR expression had no problems with rejection or functional deterioration. These findings are consistent with the theory that expression of HLA-DR antigens on renal tubular cells may be a marker of rejection and poor graft outcome. PMID- 15354974 TI - Comparison of cystatin C, creatinine and creatinine clearance vs. GFR for detection of renal failure in renal transplant patients. AB - BACKGROUND: Serum cystatin C (Scyst) has been suggested as an alternative index of glomerular filtration rate (GFR) and could be useful in renal transplant patients. METHODS: In a 60-subject cohort (40 +/- 12 years old), we compared the simultaneous measurements of Scyst, serum creatinine (Screat), creatinine clearance (Ccreat), Cockcroft and Gault's estimated clearance (Ccg) and GFR measured using inulin clearance (Cin). Receiver operating characteristic (ROC) analysis was performed using two Cin cut-off (60 and 90 mL/min/1.73 m2). RESULTS: A significant correlation was found among Cin on one hand and 1/Scyst, Ccreat, 1/Screat and Ccg on the other hand. Best fits (sensitivity/specificity) at 90 mL/min/1.73 m2 were 1.18 mg/L (0.72/0.80) for Scyst, 1.32 mg/dL (0.67/0.90) for Screat, 77 mL/min (0.80/0.70) for Ccg and 104 mL/min (0.88/0.80) for Ccreat. The areas under the ROC curves were not significantly different. CONCLUSIONS: This study provides cut-off values for Screat and Ccg for detection of renal failure in renal transplant patients. However, the results also suggest that Scyst is not a more sensitive marker than Screat or Ccg for detecting renal failure in renal transplant patients. PMID- 15354975 TI - Glomerular endothelial dysfunction in chronic kidney disease. AB - A dysfunctioning glomerular endothelium was demonstrated in chronic kidney disease (CKD) patients by means of in vitro endothelial cell cytotoxicity test and of in vivo intrarenal hemodynamic study. An enhanced endothelial cell cytotoxicity in CKD patients was 26.5 +/- 12% as compared to 0.4 +/- 1% of control. An altered intrarenal hemodynamics revealed 1) a reduction in renal plasma flow, 190 +/- 67 mL/min/1.73 m2 versus control 595 +/- 45 mL/min/1.73 m2, and in peritubular capillary flow, 149 +/- 55 mL/min/1.73 m2 versus control 479 +/- 46 mL/min/1.73 m2, 2) an elevated intraglomerular hydrostatic pressure, 55 +/ 2 mmHg versus control 51 mmHg, elevated afferent arteriolar resistance, 13184 dyne x s x cm(-5) versus control 2443 +/- 154 dyne x s x cm(5), and elevated efferent arteriolar resistance, 13591 +/- 7591 dyne x s x cm(-5) versus control 3062 +/- 177 dyne x s x cm(-5). Both enhanced endothelial cell cytotoxicity and altered intrarenal hemodynamics reflect glomerular endothelial dysfunction which is likely responsible for the renal disease progression in CKD. PMID- 15354976 TI - Homocysteine in renal transplant recipients: association with transplant duration and renal function. AB - BACKGROUND: Hyperhomocystinemia is an established risk factor for cardiovascular events and has been identified as an important cause of morbidity and mortality in renal transplant recipients. This investigation was aimed to determine the effect of age and transplant duration on serum total homocysteine (tHcy) levels in renal transplant recipients. METHODS: We analyzed serum levels of tHcy, albumin, alkaline phosphatase, alanine transferase, bilirubin, calcium, corrected calcium, cholesterol, creatinine, folate, phosphate, potassium, sodium, triglycerides, urea and vitamin B12 in 88 transplant patients (ages, 14-67 years; transplant duration, 1-252 months) and 60 control subjects. RESULTS: Our results showed significant hyperhomocystinemia in transplant patients (19.92 +/- 0.72) as compared to controls (9.28 +/- 0.25), while male subjects in both groups had significantly higher tHcy than females. There was no correlation between patients' age and serum tHcy, whereas the time after transplantation was significantly correlated with tHcy (r=0.318, P<0.01). A significant correlation was observed between tHcy and serum urea, creatinine, vitamin B12 and potassium in renal transplant patients. CONCLUSION: This study clearly demonstrated significant hyperhomocystinemia and renal impairment in transplant recipients. A time-course increase in serum tHcy during posttransplant duration warrants long term monitoring of patients for effective clinical management. PMID- 15354977 TI - Pulmonary function in uremic patients on long-term hemodialysis. AB - Twenty patients with end-stage renal failure who were on maintenance hemodialysis (HD) underwent pulmonary function testing (PFT) before and shortly after an HD session. On pre-HD PFT, the mean values of all parameters except residual volume (RV) were in the normal range. Mean RV was high (152.9%), and mean diffusing capacity of the lung for carbon monoxide (DLCO) was high-normal (110.4%). The pre HD static inspiratory (PImax) and expiratory pressures (PEmax) were much lower than normal (67.4% and 36.3%, respectively). After the HD session, repeat PFT revealed a small increase in expiratory flow rates, and a significant drop in PImax. There was a strong correlation between PImax and PEmax (r=0.567, p<0.01) at the pre- and post-HD stages, indicating that common mechanism(s) are responsible for impairment of both inspiratory and expiratory muscle strength. The well-preserved DLCO was thought to be due to the use of biocompatible dialyzer membranes. Chronic vascular congestion might be the other explanation of high DLCO. PMID- 15354978 TI - Infectious risk factors for atherosclerotic vascular disease in hemodialysis patients--Chlamydia pneumoniae but not Helicobacter pylori or cytomegalovirus is associated with increased C-reactive protein. AB - BACKGROUND: Cardiovascular disease is a major problem in patients with chronic renal failure leading to increased mortality. Several infectious agents have been implicated to be associated with atherosclerosis. We were interested to evaluate whether there is a correlation between a past infection with Chlamydia pneumoniae (Cpn), Helicobacter pylori (Hp) or cytomegalovirus (CMV) and the manifestation of a symptomatic atherosclerotic disease in patients with endstage renal failure. METHODS AND RESULTS: Patients (n=267) on hemodialysis were investigated. In 101 patients with an apparent atherosclerotic disease (case group) increased IgA levels against Cpn were found (p < or = 0.0001 vs. controls; n=33). Nearly 45% of the case group had a history of myocardial infarction (MI) (p < or = 0.0001). Prior stroke was found in about 30% of patients in the case group (p < or = 0.002). Elevated CRP levels were identified as an independent risk factor for atherosclerosis in all groups. IgA seropositivity against Cpn correlated with elevated CRP values for all atherosclerotic patients (p < or = 0.001), especially in the group of patients with MI (p < or = 0.019) and peripheral vascular disease (p < or = 0.005). There was no correlation between CMV (IgG, IgM) or Hp (IgA, IgG) seropositivity and atherosclerotic disease. CONCLUSION: IgA seropositivity for Cpn and elevated CRP values but not Hp or CMV was associated with an increased rate of symptomatic atherosclerotic manifestations as MI, stroke, cerebral or peripheral atherosclerosis in patients with endstage renal disease on hemodialysis. PMID- 15354979 TI - Improved anemia and reduced erythropoietin need by medical or surgical intervention of secondary hyperparathyroidism in hemodialysis patients. AB - BACKGROUND: The available literature is still controversial and shows that surgical (parathyroidectomy, PTX) or medical (calcitriol) treatment actually improved or even corrected the rhEPO-resistant anemia of ESRD patients with severe SHP. The aims of this study were to 1) assess the influence of SHP on hematological parameters in ESRD patients, 2) evaluate whether or not calcitriol could improve anemia and reduce the need of erythropoietin in dialysis patients, and 3) investigate the longitudinal effect of a parathyroidectomy for 6 months on regarding any improvements in calcitriol-refractory ESRD patients. METHODS: 37 chronic hemodialysis patients in Chang Gung Memorial Hospital Dialysis Unit were divided into two groups: patients with SHP (iPTH>300 pg/mL) and patients without SHP (ipTH<300 pg/mL) before calcitriol therapy was applied. Sixteen patients remain with a status of hyperparathyroidism and were considered candidates for calcitriol therapy. Furthermore, we divided the patients according to the response of HPT to calcitriol into responding patients and nonresponding patients. Among nonresponder groups, three patients agreed to accept surgical intervention to treat their hyperparathyroidism status. RESULTS: The phosphate levels and serum alkaline phosphatase levels in patients with SHP were significantly higher when compared with those without SHP (P<0.05). As for the hematological data, hematocrit for patients with SHP was significantly higher than those without SHP (10.5 +/- 0.6 vs. 8.9 +/- 0.8, p<0.05). Other hematological parameters such as transferrin saturation and serum ferritin were not significantly different. We found a significant difference in alkaline phosphate levels in responding and nonresponding patients at 6 months on calcitriol therapy. Concomitantly, the hematocrit level is significantly higher in responding group when compared to those in nonresponding group (10.63 +/- 0.72 vs. 8.96 +/- 1.21, p<0.01). As for the dose of EPO requirement, significant difference between groups was also found after 6-month treatment (3617 +/- 2011 vs. 5416 +/- 1947, p<0.05). As for rhEPO dose requirement, positive effects of PTX were significantly found. The rhEPO doses needed to maintain patients in the hematocrit target range of 30-33% decreased gradually by 29% from 5323 +/- 1326 micro to 3774 +/- 2145 micro per week. The hematocrit level showed a significant increase at 3 months after PTX (p<0.05). This effect lasted until 6 months after PTX. The serum ferritin level was constantly around 350 to 400 pg/mL. While the transferrin saturation decreased 3 months after PTX (p<0.05) and recovered at 6 months. CONCLUSION: ESRD patients with SHP, usually associated with more severe anemia show resistance to rhEPO. In this case, investigation of SHP is strongly recommended with measurement of serum PTH, phosphate and alkaline phosphatase level. Treatment of calcitriol has a beneficial effect on renal anemia in ESRD patients with SHP. In addition, PTX could also provide another choosing therapy in improving renal anemia when medical treatment fails. PMID- 15354980 TI - Intermittent hirudin versus continuous heparin for anticoagulation in continuous renal replacement therapy. AB - BACKGROUND: Besides possible bleeding complications a further problem in anticoagulation during continuous renal replacement therapy (CRRT) is the development of heparin-induced thrombocytopenia type II (HIT II) where further anticoagulation with heparin is contraindicated. The application of continuous hirudin as alternative for heparin caused bleeding complications by comparable filter efficacy. Aim of this prospective-controlled pilot study was to compare the efficacy and safety of intermittent hirudin and continuous heparin for anticoagulation during CRRT in critically ill patients. METHODS: 26 patients receiving CRRT were randomly allocated to two groups: Heparin group (14 patients): continuous administration of 250 IU/h heparin, dose was adjusted in 125 IU/h steps with a targeted activated clotting time (ACT) of 180-210 s. Hirudin group (12 patients): initial bolus application of 2-2-5 microg/kg hirudin, dose was adjusted in 2 microg/kg bolus steps with a targeted ecarin clotting time (ECT) >80 s. Observation time was 96 hours. RESULTS: Measured filter run time was virtually longer for heparin. No bleeding complications were observed in the hirudin group, two bleeding complications in the heparin group. CONCLUSIONS: Intermittent hirudin can be used safely for anticoagulation in CRRT. However, the in tendency better filter survival for heparin elucidates the need for further investigations to find the right dosage equilibrium between filter clotting and bleeding complications. PMID- 15354981 TI - Acute renal failure in the neonatal period. AB - Acute renal failure (ARF) is a common problem in the neonatal intensive care unit (NICU). In most cases, ARF is associated with a primary condition such as sepsis, metabolic diseases, perinatal asphyxia and/or prematurity. This retrospective study investigated the course of illness, therapeutic interventions, early prognosis and risk factors associated with development of ARF in the neonatal period. A total of 1311 neonates were treated in our NICU during the 42-month study period, and 45 of these babies had ARF. This condition was defined as serum creatinine level above 1.5 mg/dL despite normal maternal renal function. The data collected for each ARF case were contributing condition, cause and clinical course of ARF, gestational age and birth weight, age at the time of diagnosis, treatment, presence of perinatal risk factors and need for mechanical ventilation. The frequency of ARF in the NICU during the study period was 3.4%. Premature newborns constituted 31.1% of the cases. The mean birth weight in the group was 2863 +/- 1082 g, and the mean age at diagnosis was 6.2 +/- 7.4 days. The causes of ARF were categorized as prerenal in 29 patients (64.4%), renal in 14 patients (31.1%) and postrenal in 2 patients (4.4%). Forty-seven percent of the cases were nonoliguric ARF. Asphyxia was the most common condition that contributed to ARF (40.0%), followed by sepsis/metabolic disease (22.2%) and feeding problems (17.8%). Therapeutic interventions were supportive in 77.8% of the cases, and dialysis was required in the other 22.2%. The mortality rate in the 45 ARF cases was 24.4%. Acute renal failure of renal origin, need for dialysis, and need for mechanical ventilation were associated with significantly increased mortality (p<0.05). There were no statistical correlations between mortality rate and perinatal risk factors, oliguria, prematurity or blood urea nitrogen and creatinine levels. The study showed that, at our institution, ARF in the neonatal period is frequently associated with preventable conditions, specifically asphyxia, sepsis and feeding problems. Supportive therapy is effective in most cases of neonatal ARF. Acute renal failure of renal origin, need for dialysis, and need for mechanical ventilation were identified as indicators of poor prognosis in these infants. Early recognition of risk factors and rapid effective treatment of contributing conditions will reduce mortality in neonatal ARF. PMID- 15354982 TI - Does human ochratoxin A aggravate proteinuria in patients with chronic renal disease? AB - BACKGROUND: Ochratoxin A (OTA) is a nephrotoxic metabolite occurring in foodstuffs. In the last decade, OTA-induced nephropathy in man and animals have been confirmed by previous literature. The correlation between OTA and the severity of CRI and nephrotic syndrome was also researched. Therefore, this study was designed to determine whether OTA also played an important role in renal insufficiency of patients with chronic renal diseases in Taiwan. METHODS: The patients in this study were divided into nonnephrotic syndrome and nephrotic syndrome groups, first, to look for the relation between urine protein and OTA. And then these patients were also divided into six groups: (I) patients with chronic glomerulonephritis; (II) patients with chronic interstitial nephritis; (III) patients with diabetes mellitus; (IV) patients with hypertension; (V) patients with other diseases; (VI) patients with unknown reasons. For all groups, laboratory evaluation of kidney such as serum creatinine, urinary creatinine, creatinine clear rate, urinary protein, and urinary analysis were carried out coupled with determination of ochratoxin A level in urine. RESULTS: Higher levels of OTA were found in patients with nephrotic syndrome. There was a significantly positive correlation (P<0.001) between 24-hr OTA and 24-hr urine protein. On the other hand, the mean excretion of OTA in DM group (group III) was found significantly higher compared to the other groups (P<0.05). Distinct differences (P<0.01) were found especially when DM group was compared with patients with chronic glomerulonephritis (group I; P=0.0019), patients with chronic interstitial nephritis (group II; P=0.0032) and patients with hypertension (group IV; P=0.0062). CONCLUSION: The results could lead to the conclusion that OTA could play an important role in proteinuria of patients with chronic renal diseases in Taiwan. And OTA may play a role in diabetes patients with nephropathy. Further longitudinal study is needed to clarify the role of OTA in diabetic nephropathy. PMID- 15354983 TI - Diabetes mellitus, hyperhomocystinemia and atherosclerotic vascular disease in Taiwanese chronic hemodialysis patients: a retrospective study. AB - AIM: Diabetic patients with hemodialysis (HD) have a high mortality rate from atherosclerotic vascular disease (ASVD). However, the extent of the role of hyperhomocystinemia as a risk factor of ASVD is uncertain in diabetic HD patients. We investigated whether there was an association with ASVD events in diabetics and non-diabetics where these were chronic hyperhomocystinemia HD patients. METHODS: Two hundred patients undergoing HD were included in the study. About 50% of the patients had diabetes mellitus (DM). They had predialysis blood work performed for total homocysteine. A history of DM was elicited using information from the patients' questionnaires and verified by careful inpatient and outpatient chart review. RESULTS: A total of 196 patients had hyperhomocystinemia and were enrolled this study. Mean homocysteine concentration was 29.7 +/- 6.6 micromol/L overall. DM was present in 50.0% of patients. The mean homocysteine concentration was 29.4 +/- 9.5 micromol/L and 29.9 +/- 9.7 micromol/L in diabetic HD patients (n=98) and non-diabetic HD patients (n=98), respectively (P=0.71). There was no association with hyperhomocystinemia between diabetic and non-diabetic in chronic HD patients. There were significant differences including age, sex, HDL cholesterol, triglycerides, hypertension, smoking, serum creatinine, dialysis duration and glucose intolerance in the two groups (P<0.05). There were also significant differences in ASVD (P=0.0027) and CVD (P=0.0017) between diabetics and non-diabetics in cases of chronic hyperhomocystinemia HD patients. The adjusted odds ratio for ASVD was 3.02 (95%CI, 1.63 to 5.59) for those subjects with a DM in the highest quartile compared with the lowest 3 quartiles. CONCLUSIONS: There were associations with ASVD and CVD in diabetics and non-diabetics in cases of chronic hyperhomocystinemia HD patients. There was no association with hyperhomocystinemia between diabetic and non-diabetic in Taiwanese chronic HD patients. This study found that the presence of DM and advanced age were the major determinants for ASVD events in chronic HD patients, rather than the levels of homocysteine. PMID- 15354984 TI - Hypercalcemia in prostate cancer with positive neuron-specific enolase stain. AB - Hypercalcemia is a common complication of malignant diseases with or without bone metastasis. Hypercalcemia in prostate cancer is rarely seen. The exact mechanism of prostate cancer-related hypercalcemia is still uncertain. Secretion of parathyroid hormone-related peptides (PTH-rP) is thought to be one of the possible mechanisms. We reported a rare case of prostate cancer with hypercalcemia (13 mg/dL). Bone marrow biopsy showed metastatic adenocarcinoma. The cells were also positive for neuron-specific enolase, which is the specific marker for neuroendocrine cell. The finding suggested that the prostate cancer cell derived from the neuroendocrine cell, which might synthesize PTH-rP and be responsible for the observed hypercalcemia. PMID- 15354985 TI - Acute renal failure following para-phenylenediamine (PPD) poisoning: a case report and review. AB - We report a case of systemic poisoning with para-phenylenediamine (PPD) presenting with characteristic features of severe angioneurotic edema, rhabdomyolysis and intravascular hemolysis with hemoglobinuria culminating in acute renal failure. Though rare in western countries, such poisoning is not uncommon in East Africa, Indian subcontinent and Middle East countries. We discuss here the clinical features and key management issues of systemic PPD poisoning. PMID- 15354986 TI - Hypercalcemia and elevated serum 1.25-dihydroxyvitamin D in an end-stage renal disease patient with pulmonary cryptococcosis. AB - Hypercalcemia occurs relatively often in dialysis patients. The most common cause of hypercalcemia in dialysis patients is the conventional therapy with calcium and calcitriol. Besides, secondary hyperparathyroidism, low turnover bone diseases, and immobilization are also common causes of hypercalcemia in dialysis patients. Fungal infection associated with hypercalcemia has been infrequently reported. We describe a 71-year-old female woman with end-stage renal disease and diabetes mellitus, who developed severe hypercalcemia. Pulmonary cryptococcosis, with increased concentration of serum 1,25-dihydroxyvitamin D (1,25(OH)2D), was diagnosed. Her serum concentration of calcium and 1,25(OH)2D returned to normal after antifungal treatment. Thus, hypercalcemia was mediated by extrarenal overproduction of 1,25(OH)2D in this patient. PMID- 15354987 TI - The attention economy and the EMBS. PMID- 15354988 TI - Seeing clearly with Refractec Inc. PMID- 15354989 TI - Integrative science: biosignal processing and modeling. PMID- 15354990 TI - Celebrating the accomplishments of clinical engineering. PMID- 15354991 TI - Challenges facing independent multihospital healthcare technology management systems. PMID- 15354992 TI - Performance measures of ISM-band and conventional telemetry. PMID- 15354993 TI - Clinical engineers in the 21st century. PMID- 15354994 TI - Using a decision support system tool for healthcare technology assessments. PMID- 15354995 TI - Clinical engineers: stewards of healthcare technologies. PMID- 15354996 TI - Medical and information technologies converge. PMID- 15354997 TI - Identification of human errors during device-related accident investigations. PMID- 15354998 TI - Planning hospital medical technology management. PMID- 15354999 TI - Stress proteins are proinflammatory in vivo. PMID- 15355000 TI - Magnetobiology: a historical view. PMID- 15355001 TI - Tissue engineering latest news. PMID- 15355002 TI - Informed consent for a device investigation. PMID- 15355003 TI - Functional magnetic resonance imaging for brain mapping in neurosurgery. AB - One of the most pertinent applications of the principle primum non nocere (first do no harm) is in the optimization of neurosurgical procedures for patients with resectable lesions. The gold standard for identifying eloquent areas of the brain to be avoided in resections is direct cortical stimulation and somatosensory evoked potential monitoring, which is itself an invasive, cumbersome and difficult technique for mapping these areas. Functional magnetic resonance imaging shows great promise as a viable noninvasive alternative to invasive mapping as well as significant current clinical utility in cases in which it cannot yet fully supplant cortical stimulation methods. Ongoing work is directed toward overcoming technical limitations, improved mapping of complex functions such as language and memory, and mapping of white matter tracts. PMID- 15355004 TI - Applications and limitations of perfusion-dependent functional brain mapping for neurosurgical guidance. AB - Perfusion-dependent brain mapping modalities, such as functional magnetic resonance imaging, positron emission tomography, and optical imaging of intrinsic signals, have become increasingly popular for neurosurgical guidance because they offer a relatively rapid and noninvasive means of mapping brain function. These modalities are unique because they rely on perfusion-related signals that are coupled with neuronal activity to map the brain instead of measuring electrophysiological responses. They consequently present unique challenges to the clinician in terms of understanding the significance and limitations of the maps they produce. In particular, one must be aware of limitations of the modalities with respect to spatial specificity, sensitivity, and reliability of these maps and how the presence of intracranial lesions may further complicate these issues. The authors review the evolution, interpretation, and limitations of perfusion-based brain mapping techniques, with special attention to clinical implications of the brain maps. PMID- 15355005 TI - A locus-driven mechanism for rapid and automated atlas-assisted analysis of functional images by using the Brain Atlas for Functional Imaging. AB - OBJECT: Functional imaging is an established neurosurgical modality for studying the brain in health and disease. Identifying numerous activation loci on many functional images and reading their underlying cortical and subcortical anatomy, coordinates, and anatomical and functional values is a tedious, time-consuming, and error-prone task. In this study the authors propose a novel approach to this problem by using an electronic brain atlas in conjunction with a locus-driven mechanism. METHODS: The Brain Atlas for Functional Imaging containing an enhanced and extended electronic version of the Talairach-Tournoux brain atlas was used for analysis. It enables loading of anatomical and functional data, correlation of these data, identification of activation loci, and their labeling with Brodmann areas, gyri, and subcortical structures by means of the atlas. The Talairach proportional grid system transformation is used to register the anatomical and functional data with the atlas. The availability of numerous tools supports this process. A locus-driven mechanism for analysis of activation loci is implemented. Locus placement within the activation region is supported by thresholding, and its location can be further edited in three dimensions on any orthogonal plane. Once all loci are identified and edited, their labels, coordinates, and anatomical/functional values are read automatically and saved in an external file. This mechanism enables the analysis to be performed in an automated, rapid, explicit, three-dimensionally consistent, and user-friendly way. CONCLUSIONS: The electronic brain atlas with locus-driven mechanism is a useful tool for localization analysis of functional images. PMID- 15355006 TI - A primer on diffusion tensor imaging of anatomical substructures. AB - In this article, the authors review the application of diffusion tensor (DT) magnetic resonance (MR) imaging to demonstrate anatomical substructures that cannot be resolved by conventional structural imaging. They review the physical basis of DT imaging and provide illustrative anatomical examples. The DT imaging technique measures the self-diffusion, or random thermal motion, of the endogenous water in nerve tissue. Because of the preferred diffusion of water molecules along the nerve fiber direction, DT imaging can measure the orientation of the neural fiber structure within each voxel of the MR image. The fiber orientation information yielded by DT imaging provides a new contrast mechanism that can be used to resolve images of anatomical substructures that cannot otherwise be visualized using conventional structural imaging. The authors illustrate how DT imaging can resolve individual pathways in the brainstem as well as individual nuclei of the thalamus and conclude by describing potential applications in neurosurgery. PMID- 15355007 TI - Magnetic resonance spectroscopy of atypical diffuse pontine masses. AB - OBJECT: Diffuse pontine gliomas in children carry a dismal prognosis, with a mean survival of less than 1 year despite therapy. The diagnosis is based on the characteristic changes demonstrated on traditional magnetic resonance (MR) imaging. A few typically MR imaging-appearing pontine masses, however, do not behave in the expected fashion, which calls the original diagnosis into question. METHODS: The authors conducted a retrospective review of data obtained in 42 children (age 6 months-13 years) in whom diffuse pontine glioma had been diagnosed at their institution. Five of these patients (12%) survived longer than expected (> 18 months). There were no differences in these patients in terms of demographics, presentation, traditional imaging findings, or treatment compared with the group as a whole. Magnetic resonance spectroscopy, however, demonstrated two distinct patterns not seen in typical diffuse pontine gliomas. In two patients elevated lipid and lactate levels were shown, with decreased levels of choline, myoinositol, and N-acetyl-aspartate (NAA). In the other patients strikingly elevated choline/creatinine ratios and myoinositol levels were observed in comparison with typical pontine tumors. CONCLUSIONS: These MR spectroscopy patterns demonstrated in this retrospective study seem to convey prognostic information and may lead to an expansion of this diagnostic tool. PMID- 15355008 TI - Toward functional neuronavigation: implementation of functional magnetic resonance imaging data in a surgical guidance system for intraoperative identification of motor and language cortices. Technical note and illustrative case. AB - In recent years, surgical navigation systems have become equipped to allow incorporation of data such as functional neuronavigation data. Functional magnetic resonance (fMR) imaging is a noninvasive modality that demonstrates various brain functions. Although still in an experimental stage, fMR imaging is a promising tool for mapping of motor and language functions. One advantage is that it can be implemented in presurgical imaging protocols and is therefore potentially widely available in general neurosurgical practice. In this paper the integration of fMR imaging and surgical navigation is described, and the potential advantages and pitfalls of its application in clinical practice are discussed. PMID- 15355009 TI - Preoperative magnetic source imaging for brain tumor surgery: a quantitative comparison with intraoperative sensory and motor mapping. AB - OBJECT: The aim of this study was to compare quantitatively the methods of preoperative magnetic source (MS) imaging and intraoperative electrophysiological cortical mapping (ECM) in the localization of sensorimotor cortex in patients with intraaxial brain tumors. METHODS: Preoperative magnetoencephalography (MEG) was performed while patients received painless tactile somatosensory stimulation of the lip, hand, and foot. The early somatosensory evoked field was modeled using a single equivalent current dipole approach to estimate the spatial source of the response. Three-dimensional magnetic resonance image volume data sets with fiducials were coregistered with the MEG recordings to form the MS image. These individualized functional brain maps were integrated into a neuronavigation system. Intraoperative mapping of somatosensory and/or motor cortex was performed and sites were compared. In two subgroups of patients we compared intraoperative somatosensory and motor stimulation sites with MS imaging-based somatosensory localizations. Mediolateral projection of the MS imaging source localizations to the cortical surface reduced systematic intermodality discrepancies. The distance between two corresponding points determined using MS imaging and ECM was 12.5 +/- 1.3 mm for somatosensory-somatosensory and 19 +/- 1.3 mm for somatosensory-motor comparisons. The observed 6.5 mm increase in site separation was systematically demonstrated in the anteroposterior direction, as expected from actual anatomy. In fact, intraoperative sites at which stimulation evoked the same patient response exhibited a spatial variation of 10.7 +/- 0.7 mm. CONCLUSIONS: Preoperative MS imaging and intraoperative ECM show a favorable degree of quantitative correlation. Thus, MS imaging can be considered a valuable and accurate planning adjunct in the treatment of patients with intraaxial brain tumors. PMID- 15355010 TI - Comparison of conscious sedation and general anesthesia for motor mapping and resection of tumors located near motor cortex. AB - OBJECT: The surgical treatment of tumors located near eloquent cortex carries a high risk of inducing worsening neurological deficits. Intraoperative electrocorticography techniques have been developed to help identify these areas at the time of surgery in an effort to minimize such risks. The optimal anesthetic technique for conducting these procedures, however, has never been determined. METHODS: The authors conducted a retrospective study to compare patients who underwent intraoperative motor mapping between September 2000 and May 2002. Demographic and neurophysiological monitoring data were collected from the hospital records. Patients were divided into two groups based on the anesthetic technique used for surgery: in Group 1 general anesthesia was used, and in Group 2 conscious sedation. Group 1 comprised 24 patients (mean age 47 years) with 16 right- and eight left-sided lesions. Group 2 consisted of 21 patients (mean age 46 years) with 18 right- and three left-sided lesions. Pathological diagnoses were similar between the two groups. Motor stimulation was elicited in 12 patients (50%) in Group 1 and in 21 patients (100%) in Group 2 (p < 0.001). In addition, the mean stimulation amplitude required was significantly higher (13 mA) in patients in whom conscious sedation was used as opposed to general anesthesia (5 mA, p < 0.0001). Electrographic evidence of seizures was seen in 29% of Group 1 compared with 10% of Group 2 patients (p > 0.05). CONCLUSIONS: The use of conscious sedation as an anesthetic technique for motor mapping not only improves the chances of achieving successful stimulation and identification of motor cortex in relationship to the lesion, but it also allows for repetitive monitoring of the patient's motor function during resection of the lesion. PMID- 15355011 TI - Surgical management of unruptured posterior carotid artery wall aneurysms. AB - Intracranial aneurysms arising from the posterior wall of the supraclinoid carotid artery are extremely common lesions. The aneurysm dilation typically occurs in immediate proximity to the origin of the posterior communicating artery and, less commonly, the anterior choroidal artery (AChA). Because of the increasingly widespread use of noninvasive neuroimaging methods to evaluate patients believed to harbor cerebral lesions, many of these carotid artery aneurysms are now documented in their unruptured state, prior to occurrence of subarachnoid hemorrhage. Based on these factors, the management of unruptured posterior carotid artery (PCA) wall aneurysms is an important element of any neurosurgical practice. Despite impressive recent advances in endovascular therapy, the placement of microsurgical clips to exclude aneurysms with preservation of all afferent and efferent vasculature remains the most efficacious and durable therapy. To date, an optimal outcome is only achieved when the neurosurgeon is able to combine systematic preoperative neurovascular assessment with meticulous operative technique. In this report, the authors review their surgical approach to PCA wall aneurysms, which is greatly based on the extensive neurovascular experience of the senior author. Focus is placed on their methods of preoperative evaluation and operative technique, with emphasis on neurovascular anatomy and the significance of oculomotor nerve compression. They conclude by discussing surgery-related complications, with a particular focus on intraoperative rupture of aneurysms and their management, and the postoperative ischemic AChA syndrome. PMID- 15355012 TI - Meningiomas: the role of preoperative angiography and embolization. AB - The field of interventional neuroradiology has experienced remarkable technological developments in microcatheters and embolic materials during the past two decades. The realm of meningioma therapy has benefited handsomely from the combination of these technical improvements and the knowledge of experienced practitioners in this field. Transarterial embolization has become a standard procedure in the preoperative management of meningiomas. The authors describe the indications, pretreatment evaluation, techniques, and outcomes when preoperative angiography and embolization are performed in the treatment of these tumors. PMID- 15355013 TI - Meningiomas of the velum interpositum: surgical considerations. AB - Meningiomas of the third ventricle are a rare subtype of pineal region tumor that arise from the posterior portion of the velum interpositum, the double layer of pia mater that forms the roof of the third ventricle. The authors review the literature concerning these meningiomas and present a case in which the lesion was resected via the supracerebellar-infratentorial approach. The relationship of the tumor to the deep venous system and the splenium of the corpus callosum guides the selection of the most advantageous surgical approach. Posterior displacement of the internal cerebral veins demonstrated on preoperative imaging provides a strong rationale for use of the supracerebellar-infratentorial approach. PMID- 15355014 TI - Neurotrophic factor in the treatment of Parkinson disease. AB - OBJECT: Parkinson disease (PD) is a well-known degenerative disease resulting in the depletion of dopamine-producing neurons in the pars compacta of the substantia nigra. Adenoviral vector delivery of neurotrophic factors may provide a potential therapy for PD. The authors examined whether glial cell line-derived neurotrophic factor (GDNF) delivered via adenoviral vector (Ad-GDNF) could promote functional recovery in a rat model of PD. Additionally, they examined whether neural precursor cells (NPCs) provide the therapeutic potential of cultured neural cells for cell regeneration and replacement in PD. METHODS: All animals underwent stereotactic injection of 6-hydroxydopamine into the right substantia nigra. Eight weeks later, the rats were tested for apomorphine-induced rotational asymmetry and evaluation of explanted grafts infected with the complementary DNA for GDNF containing NPCs and NPCs alone. In the NPC cultures of embryonic rat striata, the authors found that basic fibroblast growth factor induced the proliferation of stem cells, which give rise to spheres of undifferentiated cells that generate neurons and glia. CONCLUSIONS: In this study the authors found that the reduction of apomorphine-induced rotation was more prominent in parkinsonian rats that received Ad-GDNF-treated grafts containing NPCs (61%) than in those that received grafts of NPCs alone (16%). PMID- 15355015 TI - Spinal congenital dermal sinus associated with upper thoracic meningocele. Case Report. AB - The congenital dermal sinus is an abnormal epithelium-lined sinus tract between the skin surface and deeper tissues. It occurs during neurulation when the neural groove closes to form the neural tube on Day 26 of gestation and results from a failure of neuroectoderm to separate from the cutaneous ectoderm. The most frequent location is the lumbosacral area; an upper thoracic location is quite rare. This 37-year-old man presented with headache and numbness in both arms. No specific neurological findings were observed. Physical examination revealed a dimple at T-2. Radiography and magnetic resonance imaging of the thoracic spine revealed spina bifida at T1-3, a meningocele, and a dermal sinus tract complex. The treatment approach and outcome in this unusual case are presented. PMID- 15355016 TI - Improved procedure for the oxidative cleavage of olefins by OsO4-NaIO4. AB - [reaction: see text] Oxidative cleavage of olefins by OsO(4)-NaIO(4) sometimes suffers from low yields due to the formation of side products. It is found that the addition of 2,6-lutidine can suppress the side reactions and dramatically improve the yield of this classic reaction. PMID- 15355017 TI - Synthesis of an isostere of an O-linked glycopeptide. AB - [reaction: see text] A route for the synthesis of an electrophilic, carbocyclic galactose equivalent from D-galactose is described. The strategy utilizes ring closing metathesis with Grubbs's second-generation catalyst as the key step. The galactose-derived electrophile reacted in an S(N)2 fashion with N-Boc-cysteine methyl ester to provide an alpha-galactosylserine isostere. The method was extended to the synthesis of a glycopeptide isostere. PMID- 15355018 TI - Facile and efficient synthesis of 6-(hydroxymethyl)purines. AB - [reaction: see text] A facile and efficient methodology of the synthesis of 6 (hydroxymethyl)purine derivatives (bases and nucleosides) was developed based on Pd-catalyzed cross-coupling reactions of 6-halopurines with acyloxymethylzinc iodides followed by deprotection. Several title compounds are inhibitors of adenosine deaminase and exert cytostatic activity. PMID- 15355019 TI - Facile 5-endo amidyl radical cyclization promoted by vinylic iodine substitution. AB - [reaction: see text] BF(3).OEt(2)-catalyzed atom-transfer radical addition of iodoacetamides to alkynes yielded the corresponding vinyl iodides, which upon treatment with (t)BuOCl and I(2) afforded gamma-lactam derivatives in moderate to good yield. The mechanism was proposed to be 5-endo amidyl radical cyclization, and vinylic iodine substitution provided the driving force for the cyclization. PMID- 15355020 TI - Crystallization-induced chiral inversion as the key step for synthesis of (S)-2 acetylthio-3-phenylpropanoic acid from l-phenylalanine. AB - [reaction: see text] A novel crystallization-induced chiral inversion of (S)-2 bromo-3-phenylpropanoic acid to its (R)-enantiomer with excellent enantiomeric excess (96-99%) is achieved. Optically pure (S)-2-acetylthio-3-phenylpropanoic acid can be prepared in good yield from inexpensive and commercially available l phenylalanine via diazotization/bromination, chiral inversion, and thioacetate substitution reactions. PMID- 15355021 TI - Demonstration of the feasibility of a direct solid-phase split-pool Biginelli synthesis of 3,4-dihydropyrimidinones. AB - [reaction: see text] A direct, Lewis acid-catalyzed Biginelli synthesis of 3,4 dihydropyrimidinones has been performed on high-capacity polystyrene macrobeads with a polymer O-silyl-attached N-(3-hydroxypropyl)urea. Resin-urea was first reacted separately with either 4-bromo- or 4-chlorobenzaldehyde and LiOTf in MeCN at 80 degrees C. After washing, the beads were pooled and reacted with ethyl acetoacetate and LiOTf in MeCN at 80 degrees C. Formation of only one kind of Biginelli product per bead demonstrated the feasibility of a solid-phase non Atwal two-step split-and-pool synthesis of 3,4-dihydropyrimidinones. PMID- 15355022 TI - Intramolecular [3 + 2]-cycloaddition reaction of push-pull dipoles across heteroaromatic pi-systems. AB - [reaction: see text] Push-pull dipoles generated from the Rh(II)-catalyzed reaction of diazo imides containing tethered heteroaromatic rings undergo successful [3 + 2]-cycloaddition across the 2,3-pi-bond to provide novel pentacyclic compounds in good to excellent yields in a stereocontrolled fashion. The facility of the cycloaddition is critically dependent on conformational factors in the transition state. PMID- 15355023 TI - Efficient and selective synthesis of 6,7-Dehydrostipiamide via Zr-catalyzed asymmetric carboalumination and Pd-catalyzed cross-coupling of organozincs. AB - [structure: see text] 6,7-Dehydrostipiamide has been synthesized in 23% yield in 15 steps in the longest linear sequence through the application of the Zr catalyzed asymmetric carboalumination and the Pd-catalyzed organozinc cross coupling in addition to the Brown crotylboration, the Corey-Peterson olefination, and the Corey-Fuchs reaction for carbon-carbon bond formation. PMID- 15355024 TI - Stereoselective Mn-mediated coupling of functionalized iodides and hydrazones: a synthetic entry to the tubulysin gamma-amino acids. AB - [structure: see text] Synthesis of gamma-amino acids, important building blocks in bioorganic and natural product chemistry, is accomplished using a stereoselective carbon-carbon bond construction of the chiral amine. Alkyl iodides and chiral hydrazones with protected alcohol functionality are coupled via highly diastereoselective photolytic Mn-mediated addition to the C=N bond, providing access to enantiomerically pure multifunctional chiral alpha-branched amines. Reductive N-N bond cleavage and alcohol oxidation provides alpha substituted gamma-amino acid building blocks for tubulysin D. PMID- 15355025 TI - A facile synthetic method to prepare fluorescently labeled ROMP polymers. AB - [structure: see text] To probe the activities of sperm ADAM protein (fertilinbeta), we devised a general synthetic strategy to generate fluorescently labeled fertilinbeta oligopeptide polymers. Immunofluorescence studies with these polymers demonstrated that fertilinbeta polymers bind specifically to a protein receptor on the mouse egg plasma membrane. PMID- 15355026 TI - A new family of macrocycles produced by sequential Claisen-Schmidt condensations. AB - [reaction: see text] Members of a new family of macrocycles have been synthesized in one step, from simple building blocks, by sequential Claisen-Schmidt condensations. PMID- 15355027 TI - Horning-crown macrocycles: novel hybrids of calixarenes and crown ethers. AB - [reaction: see text] Novel macrocycles possessing ether linkages and 2,6 disubstituted phenolics were produced in one step and with 100% atom economy by isoaromatization of chameleon macrocyclic precursors possessing 2,6 diarylidenecyclohexanone moieties. Intramolecular hydrogen bonding of the phenolic hydrogen atoms influenced the shape of the macrocycles and dictated host guest behavior. PMID- 15355028 TI - Microwave-assisted preparation of aryltetrazoleboronate esters. AB - [reaction: see text] The addition of azido trimethylsilane to arylnitrileboronate esters is shown to proceed rapidly in dimethoxyethane to give aryltetrazoleboronate esters in moderate to good yields, with dibutyltin oxide as catalyst. PMID- 15355029 TI - Folding propensity of cyclohexylether-delta-peptides. AB - [structure: see text] Linear (n = 2-18) and cyclic oligomers (n = 3-8) of a cyclohexylether-delta-amino acid (COA) were prepared in high yield and stereopurity. CD spectra of the linear oligomers were indicative of secondary structure formation. X-ray crystal structures of cyclic COA oligomers revealed hydrophobic packing and internal 5- and 10-membered-ring hydrogen bonds. Ether and amide oxygens reside preferably in an ap orientation. This conformational locking is apparently broken by a C-2 substituent in an asymmetric cyclotrimer, for which a zeolithe-like tubular structure was found. PMID- 15355031 TI - Parallel ligand screening on olefin mixtures in asymmetric hydroformylation reactions. AB - [reaction: see text] Herein we describe a new protocol for catalyst evaluation in asymmetric hydroformylation reactions where multisubstrate screening is performed in an array of parallel reactors. This method was successfully demonstrated using a mixture of styrene, allyl cyanide, and vinyl acetate. Using this screening methodology, a set of phosphite ligands was evaluated and led to the discovery of a bisphosphite ligand that gave 88% ee and unprecedented >100:1 branched:linear regioselectivity in asymmetric hydroformylation of vinyl acetate. PMID- 15355030 TI - Core-shell-type resins for solid-phase peptide synthesis: comparison with gel type resins in solid-phase photolytic cleavage reaction. AB - [reaction: see text] Novel core-shell-type resins with a rigid core and amino functionalized flexible shell were prepared with 2,4,6-trichloro-1,3,5-triazine (CNC) and Jeffamine ED-600 starting from 1% cross-linked aminomethyl (AM) polystyrene resins. All of the amino groups were located outside the resin beads, and the loading capacity was 0.2-0.4 mmol/g. The amount of CNC treated was a determining factor in the properties of the final resins. The core-shell-type resins showed superior performances in terms of the initial loading of amino acid and the photocleavage reaction compared to the gel-type resins. PMID- 15355032 TI - Titanium catalysis in the Ugi reaction of alpha-amino acids with aromatic aldehydes. AB - [reaction: see text] Ugi reaction between an (S)-alpha-amino acid, an aromatic aldehyde, and an isonitrile proceeds best under catalysis by TiCl(4) in MeOH. The sense of diastereoinduction is (S,S). PMID- 15355034 TI - Photoinduced atom-transfer cyclization of alpha-iodocycloalkanones bearing an allenyl side chain. AB - [reaction: see text] Irradiation of alpha-iodocycloalkanones bearing an allenyl side chain with a sunlamp effected atom-transfer cyclization to give cyclized products in good yield. A mechanism, involving radical atom-transfer cyclization accompanied by 1,5- and 1,4-hydrogen transfers, is proposed. PMID- 15355033 TI - Novel design of bicyclic beta-turn dipeptides on solid-phase supports and synthesis of [3.3.0]-Bicyclo([2,3])-leu-enkephalin analogues. AB - [structure: see text] External bicyclic beta-turn dipeptide mimetics provide an excellent design approach that can offer a rich chiral ensemble of structures with different backbone conformations. We report herein a novel design of a convergent combinatorial synthetic methodology, which is illustrated by the solid phase synthesis of a series of [3.3.0]-bicyclo([2,3])-Leu-enkephalin analogues. The reactions were optimized and the epimeric configurations were determined by 2D NMR spectroscopy. Biological assays show that these analogues have more potent delta binding affinity and bioactivity for delta vs micro opioid receptor, which may be related to the different conformations preferred by these analogues in our modeling studies. PMID- 15355035 TI - Copper-mediated cross-coupling reactions of N-unsubstituted sulfoximines and aryl halides. AB - [reaction: see text] Copper-mediated cross-coupling reactions of sulfoximines with aryl iodides and aryl bromides provide N-arylated sulfoximines in high yields. The method is complementary to the known palladium-catalyzed N-arylation and allows the preparation of N-arylated sulfoximines, which have previously been inaccessible. PMID- 15355036 TI - Enantioselective synthesis of imperanene via enzymatic asymmetrization of an intermediary 1,3-diol. AB - [reaction: see text] Using a chemoenzymatic synthetic strategy, (S)-imperanene and its (R)-enantiomer has been synthesized from vanillin in nine steps. The key step in the synthesis involves the use of Pseudomonas cepacia lipase (PS-30) to induce asymmetrization of the intermediary prochiral 1,3-diol in >97% ee. PMID- 15355037 TI - New fluorescence-quenching process through resumption of PET process induced by complexation of alkali metal ion. AB - [reaction: see text] Under acidic conditions, a fluorescence-quenching process of 1 induced by selective binding of alkali metal ions was discovered. The mechanism of this new process involves alkali metal ion-induced deprotonation of the ammonium ion to trigger resumption of a fluorescence-quenching PET process. PMID- 15355038 TI - Total synthesis of (+)-cocaine via desymmetrization of a meso-dialdehyde. AB - [reaction: see text] The total synthesis of (+)-cocaine is described. An extension of the recently reported proline catalyzed intramolecular enol-exo aldol reaction to a meso-dialdehyde provided the tropane ring skeleton directly with good enantiomeric excess. The meso-dialdehyde was prepared using a 2 azaallyllithium [3 + 2] cycloaddition to generate a cis-2,5-disubstituted pyrrolidine. Overall, the synthesis proceeded in 6.5% yield and 86% ee over 14 linear steps starting from commercially available 3-benzyloxy-1-propanol. PMID- 15355039 TI - A highly diastereoselective MgI2-mediated ring expansion of methylenecyclopropanes. AB - [reaction: see text] A highly diastereoselective MgI(2)-mediated ring expansion of methylenecyclopropane amides to functionalized pyrrolidines has been developed using chiral aromatic sulfinimines. The 2,3,4-trisubstituted pyrrolidines were isolated in generally good to excellent yields and in excellent diastereoselectivities for aromatic and heterocyclic sulfinimines. PMID- 15355040 TI - Experimental and theoretical study on the olefin metathesis of alkenyl Baylis Hillman adducts using second-generation Grubbs catalyst. AB - [reaction: see text] We have investigated the olefin metathesis from alkenyl Baylis-Hillman adducts using second-generation Grubbs catalyst. In the experiment, the ring-closing metathesis (RCM) product could not be found, while the cross-metathesis (CM) products were found. The computational studies provided consistent explanations for the experimental result. The most limiting factor for the RCM process using second-generation Grubbs catalyst is caused by the high strain and steric effect in the metallacyclobutane intermediates. PMID- 15355041 TI - A helicene-containing foldamer displaying highly solvent-dependent CD spectra. AB - [structure: see text] A m-phenylene ethynylene oligomer containing a helicene unit was synthesized to bias the twist sense of the folded helical conformation. The CD spectra of the helicene oligomer exhibited large Cotton effects that varied greatly with the solvent composition, including three separate conformational transitions. PMID- 15355042 TI - Asymmetric transfer hydrogenation in water with a supported Noyori-Ikariya catalyst. AB - [reaction: see text] The poly(ethylene glycol)-supported ruthenium precatalyst shown above is highly effective for asymmetric transfer hydrogenation of unfunctionalized aromatic ketones by HCOONa in neat water, affording fast rates, good to excellent enantioselectivities, and outstanding reusability. PMID- 15355043 TI - Stable, crystalline acenedithiophenes with up to seven linearly fused rings. AB - [structure: see text] We report the synthesis of a series of crystalline acenedithiophenes with up to seven linearly fused rings and silylethynyl substituents. These functional groups are designed to both improve solubility and enhance cofacial interactions in the solid. We discuss the crystal packing of these materials, as well as their physical properties such as oxidation potential, UV-vis absorption, fluorescence emission, and decomposition pathways. PMID- 15355044 TI - Studies toward the total synthesis of Wiedemannic acid. AB - [reaction: see text] We report a novel and efficient diastereoselective synthesis of wiedemannic acid analogue 30 in 16 steps from 7 using a tandem oxy Cope/Claisen/ene reaction as the key step. Comparison of NMR data between wiedemannic acid (1) and analogue 30 leads us to believe that the reported stereochemistry at the ring junction of 1 is incorrect. PMID- 15355045 TI - Preparation of a lipid a derivative that contains a 27-hydroxyoctacosanoic acid moiety. AB - [structure: see text] A general synthetic strategy for long-chain omega-1 hydroxy fatty acids has been developed, which employs as a key reaction step a cross metathesis between omega-unsaturated ester and 3-butene-2-ol. The resulting lipids were used for the preparation of lipid A derivatives of Rhizobium sin-1, which have the ability to inhibit the E. coli LPS-dependent synthesis of tumor necrosis factor by human monocytes. PMID- 15355046 TI - Synthesis of novel palladacycles and their application in Heck and Suzuki reactions under aerobic conditions. AB - [reaction: see text] Design and synthesis of a novel family of furancarbothioamide-based palladacycles are reported herein. These palladacycles are thermally stable, not sensitive to air or moisture, and are applied effectively in the Heck reaction of aryl halides with terminal olefins and in the Suzuki reaction of aryl halides with arylboronic acids. These reactions were performed under aerobic conditions, leading to turnover numbers (TONs) up to 1 x 10(5). PMID- 15355047 TI - Palladium-catalyzed dimerization disilylation of 1,3-butadiene with chlorosilanes. AB - [reaction: see text] 1,3-Butadiene reacted with chlorosilanes and Grignard reagents at 20 degrees C in the presence of a catalytic amount of Pd(acac)(2) to give disilylated dimers 2 regioselectively, which have two silyl groups (R(3)Si) at the 3- and 8-positions of a 1,6-octadiene skeleton. When phenyl- or allyl substituted chlorosilanes were used, coupling product was obtained stereo- as well as regioselectively, giving rise to only (E)-olefins. It is proposed that Pd ate complexes play important roles in both C-Si bond-forming processes. PMID- 15355048 TI - A general approach toward bakkanes: short synthesis of (+/-)-bakkenolide-a (fukinanolide). AB - [reaction: see text] An efficient, general, and fully stereocontrolled approach to the family of bakkanes is disclosed. This route highlights a highly diastereoselective Diels-Alder/aldol sequence to furnish the common hydrindane precursor for the synthesis of bakkanes. The projected common intermediate was transformed to the (+/-)-bakkenolide-A in a short sequence. PMID- 15355049 TI - Organic reactions in water: an efficient zinc-mediated stereoselective synthesis of (E)- and (Z)-trisubstituted alkenes using unactivated alkyl halides. AB - [reaction: see text] Treatment of the acetyl derivatives of the Baylis-Hillman adducts 3-hydroxy-2-methylene-alkanoates and 3-hydroxy-2-methylene-alkanenitriles with unactivated alkyl halides in the presence of Zn in saturated aqueous NH(4)Cl solution at room temperature afforded (2E)-2-substituted-alk-2-enoates in the first case and (2Z)-2-substituted-alk-2-enenitriles with high (Z)-selectivity in the second case. PMID- 15355050 TI - Arylmethyl radicals from arylmethoxybromodiazirines. AB - [reaction: see text] Photolytic decompositions of 3-arylmethoxy-3-bromodiazirines afford arylmethyl radicals by homolyses of the diazirines' excited states. PMID- 15355051 TI - Preparation of an amphiphilic resin-supported BINAP ligand and its use for rhodium-catalyzed asymmetric 1,4-addition of phenylboronic acid in water. AB - [reaction: see text] The axially chiral bisphosphine ligand, 2,2' bis(diphenylphosphino)-1,1'-binaphthyl (binap), was supported on a polystyrene poly(ethylene glycol) copolymer (PS-PEG) resin and was used successfully for the rhodium-catalyzed asymmetric 1,4-addition of phenylboronic acid to alpha,beta unsaturated ketones in water. PMID- 15355052 TI - Synthesis of diazepinones via intramolecular transamidation. AB - [structure: see text] The synthesis of a collection of bicyclic fused azepinones via an intramolecular beta-lactam ring-opening strategy is reported. Depending on the chirality of the various inputs, complete stereocontrol of product formation is achieved. PMID- 15355053 TI - Highly efficient chemical kinetic resolution of bishomoallylic alcohols: synthesis of (R)-sulcatol. AB - [reaction: see text] A highly efficient chemical kinetic resolution of bishomoallylic alcohols was developed when the alcohols underwent In(OTf)(3) catalyzed 3,5-oxonium-ene-type cyclization with steroidal aldehyde 2. Consistently high enantiomeric excess (up to >99%) was obtained. PMID- 15355054 TI - Synthesis of a bis-amino acid that creates a sharp turn. AB - [reaction: see text] The synthesis of a new bis-amino acid 1 is presented. This monomer is designed to create a tightly curved structure when assembled into oligomers. The monomer is demonstrated to couple to the previously developed monomer 2 through pairs of amide bonds to create a strongly bent spiro-ladder oligomer. The structure of oligomer 3 was determined in aqueous solution using two-dimensional NMR. PMID- 15355055 TI - Chemoselective cyclizations of divinyl ketones to cyclohexenones mediated by Lewis acid and base. AB - [reaction: see text] Chemoselective cyclizations of divinyl ketones to cyclohexenones mediated by a sterically demanding Lewis acid and an amine base have been accomplished under mild reaction conditions. The extension of this methodology to the synthesis of eight-membered rings is also demonstrated. PMID- 15355056 TI - ROMPgel-supported thiazolium iodide: an efficient supported organic catalyst for parallel stetter reactions. AB - [reaction: see text] A high-loading ROMPgel-supported thiazolium iodide was prepared via ROMPolymerization of the corresponding norbornene-derived monomer. The resulting ionic ROMPgel proved to be an efficient organic catalyst for Stetter reactions. The 1,4-dicarbonyl products, important intermediates in the synthesis of cyclopentenones and heterocycles, were obtained in high yields and excellent purities after minimal purification. The ROMPgel could be reused in up to four consecutive reaction cycles without significant loss of catalytic activity. PMID- 15355057 TI - New regiosymmetrical dioxopyrrolo- and dihydropyrrolo-functionalized polythiophenes. AB - [structure: see text] We present the synthesis of two N-alkylated poly(dioxopyrrolothiophene)s and two N-alkylated poly(dihydropyrrolothiophene)s with potential application in the field of conducting polymers. The polymers are synthesized from the corresponding 2,5-dibromothiophenes by an Ullmann-type polymerization and a Stille-type polymerization, respectively. The two N alkylated poly(dihydropyrrolothiophene)s are the first examples of amino functionalized polythiophenes built from regiosymmetrical thiophene monomers. PMID- 15355058 TI - Catalytic enantioselective conjugate addition of trimethylsilylacetylene to 2 cyclohexen-1-one. AB - [reaction: see text] The first example of catalytic asymmetric conjugate addition of TMS-acetylene to a cyclic alpha,beta-enone has been accomplished using the chiral bisoxazoline-Ni complex 9 as catalyst and dimethylalumino TMS-acetylide and 2-cyclohexen-1-one as reactants. PMID- 15355059 TI - Nankakurine A, a novel C16N2-type alkaloid from Lycopodium hamiltonii. AB - [sstructure: see text] A novel, fused-tetracyclic Lycopodium alkaloid, nankakurine A (1), consisting of a cyclohexane ring and a 3-aza bicyclo[3.3.1]nonane ring connected to a piperidine ring through a spiro carbon, was isolated from the club moss Lycopodium hamiltonii. The structure and relative stereochemistry were elucidated on the basis of spectroscopic data. PMID- 15355060 TI - Asymmetric synthesis of alpha-amino 1,3-dithioketals from sulfinimines (N sulfinyl imines). Synthesis of (2S,3R)-(-)-3-hydroxy-3-methylproline. AB - [reaction: see text] N-Sulfinyl alpha-amino 1,3-dithioketals are prepared in high de and good yield by treating sulfinimines with lithio-1,3-dithianes. Selective removal of the N-sulfinyl or the thioketal groups affords stable alpha-amino 1,3 dithioketals and N-sulfinyl alpha-amino ketones, respectively. This new sulfinimine-derived chiral building block is employed in the asymmetric synthesis of polyoxypeptin amino acid (2S,3R)-(-)-3-hydroxy-3-methylproline. PMID- 15355061 TI - Highly enantioselective dimerization of alpha,beta-enones catalyzed by a rigid quaternary ammonium salt. AB - [reaction: see text] The chiral quaternary ammonium salt 1 served as a phase transfer catalyst for the enantioselective dimerization of alpha,beta-enones, providing a route for the asymmetric syntheses of chiral 1,5-dicarbonyl compounds and alpha-alkylated gamma-keto acids. PMID- 15355062 TI - Stereoselective synthesis of the gamma-lactam hydrolysate of the thiopeptide cyclothiazomycin. AB - [reaction: see text] Bohlmann-Rahtz pyridine synthesis of a chiral nonracemic enamine and thiazolylpropynone gives a terminal-protected pyridine-containing gamma-amino acid in high optical purity in a sequential one-pot multicomponent reaction that proceeds with total control of regiochemistry and with minimal racemization. Further elaboration has established the synthesis of the gamma lactam acidic hydrolysate of the macrocyclic thiopeptide antibiotic cyclothiazomycin, a selective renin inhibitor, in only four steps and 30% overall yield and has confirmed its structure. PMID- 15355063 TI - Synthesis of fused arylboronic esters via cobalt(0)-mediated cycloaddition of alkynylboronates with alpha,omega-diynes. AB - [reaction: see text] Co(2)(CO)(6)-complexed alkynyl pinacolborane derivatives are readily transformed with functional group tolerance into fused arylboronates via the [2 + 2 + 2]cycloaddition to alpha,omega-diynes. PMID- 15355064 TI - A 1,3-Diaza-Claisen rearrangement that affords guanidines. AB - [reaction: see text] N-Alkyl-N'-tosylthioureas activated by EDCI react with azanorbonenes at room temperature through a 1,3-diaza-Claisen rearrangement, affording highly substituted, bicyclic guanidines in moderate to good yields. PMID- 15355065 TI - Synthesis of diindolocarbazoles by Ullmann reaction: a rapid route to ladder oligo(p-aniline)s. AB - [structure: see text] New and facile synthesis of symmetric diindolocarbazoles was developed using the copper-catalyzed Ullmann reaction. The key step is a double-intramolecular cyclization reaction realized on N-alkyl-3,6-dibromo-2,7 bis(2'aminophenyl)carbazole derivatives which offers the desired symmetric ladder oligo(p-aniline)s. Depending upon the nature of the side- and/or end-groups, well defined thin films and/or semiladder polymers could be obtained. These electroactive ladder oligomers may have great potential in organic electronics. PMID- 15355066 TI - Spirodihydantoin is a minor product of 5-hydroxyisourate in urate oxidation. AB - [reaction: see text] Spirodihydantoin is a minor product from oxidation of uric acid ( approximately 0.15% yield), while spiroiminodihydantoin is a major product from oxidation of 8-oxo-7,8-dihydroguanine (37% yield, pH 10.2). High pH and temperature favor the formation of both spiro compounds. (18)O labeling experiments and in situ generation and decomposition of 5-hydroxy-N7 methylisouric acid indicate that spirodihydantoin and allantoin and spiroiminodihydantoin and guanidinohydantoin are products of 5-hydroxyisourate and 5-hydroxy-8-oxo-7,8-dihydroguanine intermediates, respectively. PMID- 15355067 TI - Azoles as Suzuki cross-coupling leaving groups: syntheses of 6-arylpurine 2' deoxynucleosides and nucleosides from 6-(imidazol-1-yl)- and 6-(1,2,4-triazol-4 yl)purine derivatives. AB - [reaction: see text] 6-(Imidazol-1-yl)-, 6-(benzimidazol-1-yl)-, and 6-(1,2,4 triazol-4-yl)purine nucleosides undergo a nickel-mediated C-C cross-coupling of azole-substituted purine derivatives with arylboronic acids to give good yields of 6-arylpurine nucleosides. PMID- 15355068 TI - Chiral norbornadienes as efficient ligands for the rhodium-catalyzed asymmetric 1,4-addition of arylboronic acids to fumaric and maleic compounds. AB - [reaction: see text] A rhodium-catalyzed asymmetric 1,4-addition of arylboronic acids to fumaric and maleic compounds has been developed. While phosphorus-based chiral ligands fail to induce high stereoselectivity, chiral norbornadiene ligands have proved to be uniquely effective to achieve high enantioselectivity in these 1,4-addition reactions. PMID- 15355069 TI - Enzyme activity fingerprinting with substrate cocktails. AB - In the postgenomic era, emphasis is shifting from protein identification to protein functional analysis. Enzyme function can be characterized by measuring activity across series of substrates, which generates an activity profile or fingerprint. Activity fingerprinting is particularly useful to differentiate closely related enzymes. Previously reported fingerprinting methods use series of parallel measurements, which are complex and difficult to reproduce. Here we report a new method for fingerprinting enzyme activities based on using mixtures of substrates, or substrate cocktails, in a single reaction that is then analyzed by HPLC. The fingerprints produced are highly reproducible and allow functional differentiation and classification of closely related enzymes, as demonstrated for a series of lipases and esterases. The method is practical, general, and flexible in terms of reaction conditions and can be adapted to any reaction type. PMID- 15355070 TI - Adenine ribbon with Watson-crick and Hoogsteen motifs as the "double-sided adhesive tape" in the supramolecular structure of adenine and metal carboxylate. AB - The cocrystals of adenine and metal (II) quinoline-2-carboxylates (M = Mn2+, Fe2+, Co2+) have been obtained by self-assembly. The complexes are composed of adenine ribbons with the AA22 pairing pattern involving both Watson-Crick and Hoogsteen faces in hydrogen bonding and the neutral molecules of carboxylate positioned in inorganic layers. The very compact supramolecular structure is made by the extensive system of hydrogen bonds and face-to-face pi-pi interactions. PMID- 15355071 TI - Highly efficient, one-step macrocyclizations assisted by the folding and preorganization of precursor oligomers. AB - Highly efficient, one-step macrocyclizations leading to the formation of macrocyclic hexa(aramides) in high yields (69-82%) are described. The one-step macrocyclizations were facilitated by the preorganization or folding of the backbones of uncyclized precursors in the course of macrocyclization. The preorganization of backbones was achieved by the presence of localized three centered hydrogen bonds that were adopted in the design of a class of closely related, backbone-rigidified foldamers. The macrocyclization involved reactions between diacid chloride 1 and diamine 2. The crude reaction mixtures and products were conveniently examined by mass spectrometric method (MALDI-TOF). Compared to most traditional one-step macrocyclizations that usually require high dilution conditions and often lead to very low overall yields of the desired products, cyclic hexamers 3 were obtained as the overwhelmingly major product under a variety of reaction conditions, suggesting the generality of this approach. PMID- 15355072 TI - A new artificial cyclase for polyprenoids: enantioselective total synthesis of ( )-chromazonarol, (+)-8-epi-puupehedione, and (-)-11'-deoxytaondiol methyl ether. AB - This paper describes a new artificial cyclase, optically pure 3-o-fluorobenzyloxy 2-hydroxy-2'-(p-methoxybemzyl)-1,1'-binaphthyl.SnCl4, which is effective for the enantioselective cyclization of 2-(polyprenyl)phenol derivatives to afford polycyclic terpenoids bearing a chroman skeleton such as (-)-chromazonarol, (+)-8 epi-puupehedione, a key synthetic intermediate of (+)-wiedendiol, and (-)-11' deoxytaondiol methyl ether. PMID- 15355073 TI - Direct time-resolved and spatially resolved monitoring of molecular transport in a crystalline nanochannel system. AB - Confocal Raman microspectrometry has been applied successfully as an in situ probe of the transport of guest molecules through the one-dimensional channel system in a crystalline inclusion compound, yielding insights into the spatial distribution of guest molecules and, in particular, the variation in the spatial distribution of the guest molecules as a function of time during the transport process. PMID- 15355074 TI - Chromium(VI) binding to functionalized silica/water interfaces studied by nonlinear optical spectroscopy. AB - Organic adlayers can significantly alter the interactions of environmentally relevant surfaces with their surroundings. We present the first second harmonic and broadband sum frequency generation (SHG and BBSFG) study that illustrates how organic surface functional groups can control the mobility of the priority pollutant chromium(VI) in soil: Cr(VI) binds to ester- and acid-functionalized surfaces but not to alkane-functionalized surfaces. The implications with respect to toxic metal transport across organic adlayers at liquid-solid interfaces are that aqueous Cr(VI) can be retained by polar groups common in biopolymers but not by hydrophobic groups common in surfactants. PMID- 15355075 TI - Micellar cobaltporphyrin nanorods in alcohols. AB - Cobalt(II) meso-tetrakis(4-hexadecylamidophenyl)porphyrin self-assembles in ethanol/1-propanol 2/1 (v/v) to form a rodlike micelle with nanoscale dimensions; the nanorod is a face-to-face aggregate having a hydrophobic corona around a polar core and is thus characterized as a reverse micelle. PMID- 15355076 TI - Bidentate NHC-based chiral ligands for efficient Cu-catalyzed enantioselective allylic alkylations: structure and activity of an air-stable chiral Cu complex. AB - Cu-catalyzed addition of alkylzinc reagents to a range of allylic phosphates is promoted efficiently and with high enantioselectivity to afford tertiary as well as quaternary carbon centers (up to 98% ee). Reactions proceed to completion with 0.5-5 mol % catalyst loading and are best promoted by commercially available CuCl2.2H2O. The X-ray structure of the chiral NHC-Ag(I) complex used in the study as well as that of a catalytically active NHC-Cu(II) complex are also reported; both complexes are air-stable and are formed in >/=95% isolated yield. The isolated Cu complex, which can be handled in air, is catalytically active. The present report provides the first precedent for efficient Cu-catalyzed allylic alkylations with chiral NHC ligands. PMID- 15355077 TI - Polymer-enzyme conjugates can self-assemble at oil/water interfaces and effect interfacial biotransformations. AB - Native water-soluble enzymes were transformed into interface-binding enzymes via conjugation with hydrophobic polymers, thus enabling interesting interfacial biocatalysis between immiscible chemicals at oil/water interfaces. Such interfacial biocatalysis demonstrated a significantly improved catalytic efficiency as compared to traditional biphasic reactions with enzymes contained in the bulk aqueous phase. Particularly, polystyrene-conjugated beta galactosidase showed a catalytic efficiency that was more than 145 times higher than that of the native enzyme for a transgalactosylation reaction. It is believed that the improved accessibility of the biocatalysts to chemicals held in both phases across the interface is the key driver for the enhancement of enzyme activity. PMID- 15355078 TI - Highly selective and simple synthesis of C(2)(m)--X--C(2)(n) fullerene dimers. AB - Energetic-radiation-induced dimerization reaction of fullerenes was found to be a simple and highly selective method for synthesis of C2m-X-C2n (m = n or m not equal n) type molecules without formation of other products. Utilizing the new method, C70-C-C70, C60-C-C70, C60-C-C60, and C70-O-C70 were prepared and characterized. The method is capable of synthesizing new C2m-X-C2n molecules by introducing X (different atoms) into the reaction system. Energetic radiation created reactive sites for covalently bonded bridges between fullerene molecules originally only weakly bound by van der Waals force. This observation may open a new subject and practicable approach for polymer sciences of fullerenes. PMID- 15355079 TI - Positioning isolation and biochemical analysis of single DNA molecules based on nanomanipulation and single-molecule PCR. AB - Recently, the isolation and biochemical analysis of DNA at the single-molecule level has been recognized as very important for genetic research and clinical analysis. A unique technique for the positioning, dissection, and isolation of single DNA molecules using atomic force microscopy (AFM) has been demonstrated. Full-length genome DNA molecules were first deposited and stretched by a modified "molecular combing" technique onto a 3-aminopropyl triethoxysilane-coated mica substrate. A single DNA fragment was dissected from one of those genome DNA strands with the AFM tip at the desired position, and then isolated (or picked up) after a special operation called "kneading". All the operations including imaging, dissection, and isolation could be carried out with one tip. The isolated DNA fragment on the AFM tip could be successfully amplified by single molecule PCR. PMID- 15355080 TI - Pd(0)-catalyzed amphiphilic activation of bis-allyl alcohol and ether. AB - Pd(0).Et3B catalyzes amphiphilic activation of symmetric allylic diol 1 to promote electrophilic allylation at the alpha-position of aldehydes and nucleophilic allylation at the aldehyde CO, furnishing 3-methylenecyclopentalols 2 and thus generation of a zwitterionic trimethylenemethane species from the commercially available diol 1. PMID- 15355081 TI - Boolean logic functions of a synthetic peptide network. AB - Living cells can process rapidly and simultaneously multiple extracellular input signals through the complex networks of evolutionary selected biomolecular interactions and chemical transformations. Recent approaches to molecular computation have increasingly sought to mimic or exploit various aspects of biology. A number of studies have adapted nucleic acids and proteins to the design of molecular logic gates and computational systems, while other works have affected computation in living cells via biochemical pathway engineering. Here we report that de novo designed synthetic peptide networks can also mimic some of the basic logic functions of the more complex biological networks. We show that segments of a small network whose graph structure is composed of five nodes and 15 directed edges can express OR, NOR, and NOTIF logic. PMID- 15355082 TI - Inactivation of the carbamoyltransferase gene refines post-polyketide synthase modification steps in the biosynthesis of the antitumor agent geldanamycin. AB - The post-polyketide synthase modification of geldanamycin (1) biosynthesis is of interest as a means of introducing structural diversity into the compound. From the inactivation of a gene encoding carbamoyltransferase, we demonstrated that the C-17 hydroxylation and the C-21 oxidation precede O-carbamoylation and that the hypothetical progeldanamycin does not possess a double bond at C-4 and C-5. More importantly, our result revealed new intermediates 4,5-dihydro-7-O descarbamoyl-7-hydroxygeldanamycin (3) and 4,5-dihydrogeldanamycin (5), indicating that O-carbamoylation occurs prior to the C-4,5 cis double bond formation in geldanamycin biosynthesis. PMID- 15355083 TI - Identification of an adsorption complex between an alkane and zeolite active sites. AB - Direct observation of the Bronsted acid site signal in an active zeolite catalyst following adsorption of stoichiometric quantities of isobutane reveals the presence of a specific adsorption complex. Independent polarization transfer experiments in which magnetization originates with either the catalyst or the adsorbed isobutane confirm this assignment. The initial steps in alkane reactivity are poorly defined, and this experimentally verified complex is proposed as a route to C-H bond activation in solid acids. PMID- 15355084 TI - Chemoselective silylzincation of functionalized terminal alkynes using dianion type zincate (SiBNOL-Zn-ate): regiocontrolled synthesis of vinylsilanes. AB - A regio- and chemoselective silylzincation reaction of various functionalized alkynes was developed using a newly designed dianion-type zincate. Silylzincation of terminal alkynes, followed by electrophilic trapping, proved to be a powerful tool for the one-pot, regiocontrolled generation of trisubstituted functionalized alkenes. The functionalized vinylzincate intermediate also undergoes copper- and palladium-catalyzed C-C bond-forming reactions in high yields and with high regio and chemoselectivities. PMID- 15355085 TI - Catalytic enantioselective Michael addition of 1,3-dicarbonyl compounds to nitroalkenes catalyzed by well-defined chiral ru amido complexes. AB - Well-defined chiral Ru amido complexes promoted asymmetric Michael addition of 1,3-dicarbonyl compounds including malonates, beta-keto esters, and 1,3-diketones to nitroalkenes to give the corresponding adducts with excellent ees and in excellent yields. PMID- 15355086 TI - Diastereoselective intramolecular temporary silicon-tethered rhodium-catalyzed [4+2+2] cycloisomerization reactions: regiospecific incorporation of substituted 1,3-butadienes. AB - Transition metal-catalyzed carbocyclization reactions represent powerful methods for the construction of complex polycyclic systems. We have developed a regiospecific and diastereoselective intramolecular temporary silicon-tethered rhodium-catalyzed [4+2+2] cycloisomerization reaction of a tethered enyne for the construction of tricyclic eight-membered heterocycles and carbocycles. This methodology also allows (E)- and (Z)-olefins to be utilized in a stereospecific manner. The incorporation of either alkene geometry is particularly significant, since related carbocyclization reactions are often limited in this respect. Finally, the ability to utilize carbon-tethered 1,6-enynes and to regiospecifically incorporate substituted 1,3-butadiene derivatives provides exciting opportunities for future applications toward the total synthesis of cyclooctanoid containing diterpenes. PMID- 15355087 TI - Multicatalytic processes using diverse transition metals for the synthesis of alkenes. AB - A series of cascade processes for the synthesis of alkenes from alcohols is described. Each individual step is catalyzed with a specific transition metal complex. The oxidation-methylenation one-pot procedure took place in the presence of a palladium and a rhodium catalyst to produce the desired terminal alkenes in high yields. A methylenation-ring-closing metathesis allowed the synthesis of cyclic alkenes from carbonyl derivatives, using the second-generation metathesis catalyst. Finally, an oxidation-methylenation-RCM process that involves up to three different transition metal catalysts in the same vessel is presented. PMID- 15355088 TI - Strong solvatochromic fluorescence from the intramolecular charge-transfer state created by excited-state intramolecular proton transfer. AB - In this work, we report a peculiar positive solvatochromism in the keto emission of the acceptor-substituted 2-(2'-hydroxyphenyl)benzoxazoles (HBO), which originates from the excited-state intramolecular proton transfer (ESIPT) followed by the intramolecular charge transfer (ICT) and subsequent solvent relaxation. This transient evolution of enhanced ICT characteristic triggered by ESIPT, which is first observed in this work, is responsible for the novel concept of a fast hyperpolarizability modulator as well as the unique solvatochromic behavior. PMID- 15355089 TI - Epoxide hydrolase-catalyzed enantioselective synthesis of chiral 1,2-diols via desymmetrization of meso-epoxides. lzhao@diversa.com. AB - The discovery, from nature, of a diverse set of microbial epoxide hydrolases is reported. The utility of a library of epoxide hydrolases in the synthesis of chiral 1,2-diols via desymmetrization of a wide range of meso-epoxides, including cyclic as well as acyclic alkyl- and aryl-substituted substrates, is demonstrated. The chiral (R,R)-diols were furnished with high ee's and yields. The discovery of the first microbial epoxide hydrolases providing access to complementary (S,S)-diols is also described. PMID- 15355090 TI - Water-soluble, exfoliated, nonroping single-wall carbon nanotubes. AB - The scalable superacid solvent, radical-initiated aryldiazonium functionalization process produces individual SWNTs without the need for surfactant wrapping, centrifugation, or sonication. This work provides a facile pathway to aryl sulfonic acid-functionalized SWNTs that are not roped or bundled, and the functionalized nanotubes are water soluble. PMID- 15355091 TI - Synthesis and reactivity of dimethyl platinum(IV) hydrides in water. AB - New hydrophilic ligands of the di(2-pyridyl)methanesulfonate family, L = dpms and Me-dpms, enable the synthesis of methyl platinum(IV) hydrides, LPtMe2H, the study of very fast CH reductive coupling, and reductive elimination of these complexes in water. In dichloromethane solutions, 13CH4 reacts with (Me-dpms)PtMe2H to produce isotopomeric complexes. PMID- 15355092 TI - An efficient [4 + 2 + 1] entry to seven-membered rings. AB - A new nickel-catalyzed procedure for the [4 + 2 + 1] cycloaddition of trimethylsilyl diazomethane with alkynes tethered to dienes has been developed. A broad range of unsaturated substrates participate in the sequence, and stereoselectivities are generally excellent. Three possible mechanisms are proposed, and each involves the generation of a transient nickel carbene species. PMID- 15355093 TI - AuCl(3)-catalyzed synthesis of highly substituted furans from 2-(1-alkynyl)-2 alken-1-ones. AB - Highly substituted furans have been synthesized by the reaction of 2-(1-alkynyl) 2-alken-1-ones and various nucleophiles under very mild reaction conditions in good to excellent yields. Gold and some other transition metals are efficient catalysts for this reaction. PMID- 15355094 TI - Electrocatalytic reduction of ROOH by iron porphyrins. AB - Electrocatalytic reduction of a series of chemical oxidants of different power (tert-butyl hydroperoxide, potassium peroxomonosulfate, peracetic acid, and m chloroperbenzoic acid) at iron-porphyrin-modified graphite electrodes is studied in buffered aqueous solutions by rotating disk and ring-disk voltammetry. Both ferric and ferrous porphyrins are catalytically active. Turnover of ferric catalysts is slower than that of the ferrous analogues and involves competing catalytic reduction and disproportionation. The kinetic data are consistent with reactant binding being the rate-determining step in catalysis by Fe(III). In catalysis by Fe(II), the turnover is controlled by the first electron transfer. The covalently linked proximal imidazole ligand is found to be crucial for achieving the Fe(III) catalysis. PMID- 15355095 TI - A pentacene with a 144 degrees twist. AB - 9,10,11,20,21,22-Hexaphenyltetrabenzo[a,c,l,n]pentacene (1) was prepared by the reaction of 1,3-diphenylphenanthro[9,10-c]furan with the bisaryne equivalent generated from 1,2,4,5-tetrabromo-3,6-diphenylbenzene in the presence of n butyllithium, followed by deoxygenation of the double adduct with low-valent titanium. The X-ray structure of 1 shows it to be the most highly twisted polycyclic aromatic hydrocarbon known, with an end-to-end twist of 143.6 degrees . Compound 1 was resolved by chromatography on a chiral support, and the pure enantiomers have specific rotations in excess of 7000 degrees , but the molecule racemizes slowly at 25 degrees C (t1/2 = 9.3 h, DeltaGrac = 23.8 kcal/mol). PMID- 15355096 TI - Alkyne and alkene complexes of a d(0) zirconocene aryl cation. AB - The generation and properties of nonchelated Zr-aryl-alkyne and Zr-aryl-alkene complexes that are stabilized by the presence of beta-Si-substituents in the alkyne and alkene ligands and fluorination of the aryl ligand are described. Reaction of [Cp'2Zr(OtBu)(ClCD2Cl)][B(C6F5)4] (1, Cp' = C5H4Me) with alkyne and alkene substrates (L) generates Cp'2Zr(OtBu)(L)+ adducts (L = HCCCH2SiMe3 (2); H2C=CHCH2SiMe3 (3); HCCMe (4); H2C=CHCH2CMe3 (5)). Equilibrium constants for substrate binding (Keq = [Zr-L][1]-1[L]-1; CD2Cl2, -89 degrees C) are much larger for the beta-Si-substituted compounds 2 (1.0(2) x 105 M-1) and 3 (1.7(4) x 103 M 1) than for hydrocarbon analogues 4 (3.6(7) x 102 M-1) and 5 (1.9(1) M-1), which is ascribed to beta-Si stabilization of the partial positive charge on Cint of the bound substrate. [Cp2Zr(C6F5)][B(C6F5)4] (7, Cp = C5H5) was generated by the reaction of Cp2Zr(C6F5)Me with [Ph3C][B(C6F5)4] in C6D5Cl. Reaction of 7 with alkyne and alkene substrates (L) generates Cp2Zr(C6F5)(L)+ adducts (L = HCCCH2SiMe3 (8); H2C=CHCH2SiMe3 (10)). No insertion of the substrate into the Zr C6F5 bond is observed in 8 (at -38 degrees C) or 10 (up to 22 degrees C). The allyltrimethylsilane ligand in 10 undergoes nondissociative alkene face exchange ("alkene flipping", i.e., exchange of the Cp2Zr(C6F5)+ unit between the two alkene enantiofaces without alkene dissociation), with a first-order rate constant kflip = 23(1) s-1 (C6D5Cl, -38 degrees C). 10 also undergoes slower reversible decomplexation of the alkene (kdissoc = 5.0(8) s-1; C6D5Cl, -38 degrees C). PMID- 15355097 TI - Backbone thioester exchange: a new approach to evaluating higher order structural stability in polypeptides. AB - An amide bond has been replaced by a thioester in bovine pancreatic polypeptide (bPP) to allow rapid and reversible (dynamic) exchange of the alpha-helical segment with other thiols in solution. We have begun to study the higher order structural stability of bPP by measuring the equilibrium constant of the "backbone thioester exchange" (BTE) reaction. The extent to which the equilibrium (KBTE) favors one set of peptides over the other, which can be easily measured, can be directly correlated to the energy gained from favorable noncovalent interactions that occur between peptide segments on either side of the thioester bond (Kfold). PMID- 15355098 TI - The earliest events in protein folding: a structural requirement for ultrafast folding in cytochrome C. AB - The folding dynamics of reduced cytochrome c (redcyt c) obtained from tuna heart, which contains a tryptophan residue at the site occupied by His33 in horse heart cytochrome c, was studied using nanosecond time-resolved optical rotatory dispersion spectroscopy. As observed previously for horse heart redcyt c, two time regimes were observed for secondary structure formation in tuna redcyt c: a fast (microseconds) and a slow (milliseconds) phase. However, the fast phase of tuna redcyt c folding was much slower and smaller in amplitude than the same phase in horse. The differences in the fast folding phases suggest that for horse heart redcyt c, the conformers that undergo the fastest observed folding have the His18-Fe-His33 heme configuration, which appears to be necessary, but not sufficient, to poise an unfolded chain conformation for fastest folding in redcyt c. PMID- 15355099 TI - Flow linear dichroism to probe binding of aromatic molecules and DNA to single walled carbon nanotubes. AB - Structures of carbon nanotube/ligand complexes were studied by flow linear dichroism (the differential absorption of light polarized parallel and perpendicular to the flow orientation direction) with the aim of establishing linear dichroism as a technique to study such systems. Anthracene, naphthalene, and DNA were chosen as ligands, and the potential for flow linear dichroism to probe ligands noncovalently (as well as covalently) bound to single-walled nanotubes is reported. Linear dichroism enables the determination of approximate orientations of the ligands on the carbon nanotubes. PMID- 15355100 TI - Site-specific synthesis and properties of oligonucleotides containing C8 deoxyguanosine adducts of the dietary mutagen IQ. AB - The site-specific synthesis of oligonucleotides containing the C8-deoxyguanosine adduct of the highly mutagenic heterocyclic amine 2-amino-3-methylimidazo[4,5 f]quinoline (IQ) has been achieved, and the oligonucleotides were characterized by UV melting temperature analysis, circular dichroism, and UV absorption spectroscopy. Examination of these data indicated that the IQ-adduct is accommodated in dramatically different environments. This sequence-dependent conformational preference is likely to play a key role in the mutagenicity and repair of IQ-modified oligonucleotides. PMID- 15355101 TI - Aromaticity of organic heterocyclothiazenes and analogues. AB - Members of a series of carbon-poor sulfur-nitrogen heterocycles and polycycles are shown by direct ab initio ipsocentric calculation to support diatropic ring currents and hence to be aromatic on the basis of magnetic criteria. They include 7-cycles S(3)N(2)(CH)(2), S(3)N(3)(CH), and S(3)N(4) and 8-cycles S(2)N(4)(CH)(2) and S(2)N(2)(CH)(4), all with 10 pi electrons. The unknown trithiatetrazepine S(3)N(4) is predicted to be at least as aromatic as its known diaza and triaza homologues. Angular-momentum arguments show that the pi-electron-rich nature of (4n + 2) SN heterocycles is the key to their diatropic current. The Woodward dithiatetrazocine parent framework S(2)N(4)(CH)(2) supports a diatropic ring current, as does its analogue in which N and CH groups are formally exchanged. Formal expansion of (4n + 2)-pi carbocyclic systems by insertion of NSN motifs in every CC bond is predicted to lead to structures that support diatropic ring currents: explicit ab initio calculation of magnetic response predicts the 24 center, 30-pi-electron heterocycle S(6)N(12)(CH)(6), formally derived from benzene, to be aromatic on the basis of this criterion. PMID- 15355102 TI - A transition state analogue for an RNA-editing reaction. AB - Deamination at C6 of adenosine in RNA catalyzed by the ADAR enzymes generates inosine at the corresponding position. Because inosine is decoded as guanosine during translation, this modification can lead to codon changes in messenger RNA. Hydration of 8-azanebularine across the C6-N1 double bond generates an excellent mimic of the transition state proposed for the hydrolytic deamination reaction catalyzed by ADARs. Here, we report the synthesis of a phosphoramidite of 8 azanebularine and its use in the preparation of RNAs mimicking the secondary structure found at a known editing site in the glutamate receptor B subunit pre mRNA. The binding properties of analogue-containing RNAs indicate that a tight binding ligand for an ADAR can be generated by incorporation of 8-azanebularine. The observed high-affinity binding is dependent on a functional active site, the presence of one, but not the other, of ADAR2's two double-stranded RNA-binding motifs (dsRBMs), and the correct placement of the nucleoside analogue into the sequence/structural context of a known editing site. These results advance our understanding of substrate recognition during ADAR-catalyzed RNA editing and are important for structural studies of ADAR.RNA complexes. PMID- 15355103 TI - Thermodynamics of folding, stabilization, and binding in an engineered protein- protein complex. AB - We analyzed the thermodynamics of a complex protein-protein binding interaction using the (engineered) Z(SPA)(-)(1) affibody and it's Z domain binding partner as a model. Free Z(SPA)(-)(1) exists in an equilibrium between a molten-globule-like (MG) state and a completely unfolded state, wheras a well-ordered structure is observed in the Z:Z(SPA)(-)(1) complex. The thermodynamics of the MG state unfolding equilibrium can be separated from the thermodynamics of binding and stabilization by combined analysis of isothermal titration calorimetry data and a separate van't Hoff analysis of thermal unfolding. We find that (i) the unfolding equilibrium of free Z(SPA)(-)(1) has only a small influence on effective binding affinity, that (ii) the Z:Z(SPA)(-)(1) interface is inconspicuous and structure based energetics calculations suggest that it should be capable of supporting strong binding, but that (iii) the conformational stabilization of the MG state to a well-ordered structure in the Z:Z(SPA)(-)(1) complex is associated with a large change in conformational entropy that opposes binding. PMID- 15355104 TI - T7 RNA polymerase transcription with 5-position modified UTP derivatives. AB - Seven UTP derivatives modified at the 5-position through an amide linkage were tested as substrates for T7 RNA polymerase (T7 RNAP) transcription. All UTP derivatives gave good yields of full-length transcript even from DNA templates that showed a significant number of abortive transcripts using unmodified UTP. A kinetic assay to determine the relative K(m) and V(max) for T7 RNAP transcription gave surprisingly similar values for UTP and the 5-position hydrophobic modifications phenyl, 4-pyridyl, 2-pyridyl, indolyl, and isobutyl. The 5-position modifications imidazole and amino, which could both be positively charged, gave K(m) values significantly higher than UTP. All seven UTP derivatives gave relative V(max) values similar to UTP, indicating that insertion of these modified bases into the transcript did not impede its elongation. PMID- 15355105 TI - Modeling branched polyethylene: copolymers possessing precisely placed ethyl branches. AB - A structural investigation of precise ethylene/1-butene (EB) copolymers has been completed using step polymerization chemistry. The synthetic methodology needed to generate four model copolymers is described; their primary and higher level structure is characterized. The copolymers possess an ethyl branch on every 9th, 15th, and 21st carbon along the backbone of linear polyethylene. Melting points and heats of fusion decrease with increased branch frequency. Differential scanning calorimetry and infrared spectroscopy show highly disordered crystal structures favoring ethyl branch inclusion. On the other hand, the EB copolymers contain high concentrations of kink and gauche defects independent of branch frequency. These model copolymers are compared with random copolymers produced using traditional chain chemistry and previously synthesized ADMET EP copolymers. PMID- 15355106 TI - Mineral-assisted pathways in prebiotic synthesis: photoelectrochemical reduction of carbon(+IV) by manganese sulfide. AB - Photoelectrochemistry on mineral surfaces has the potential to play a central role in the prebiotic syntheses of building blocks for biomolecules. In this study, photoreduction of C(+IV) as bicarbonate is used as a probe to investigate the photoelectrochemical properties of alabandite (MnS) colloidal particles. Our experimental results show that photoreduction occurs and that formate is the initial photoproduct. A quantum efficiency of 4.2% is obtained (pH = 7.5). The quantum efficiency is temperature-independent from 298 to 328 K. In addition to formate, longer chain carbon products are also produced. Ion chromatography shows the presence of acetate and propionate. Infrared spectroscopy and mass spectrometry indicate the formation of longer chain organic molecules that contain oxygenated functional groups. Our results suggest that some prebiotic syntheses could have occurred via photoelectrochemical reactions on semiconducting minerals. PMID- 15355107 TI - Total synthesis of macquarimicins using an intramolecular Diels-Alder approach inspired by a biosynthetic pathway. AB - A total synthesis of the macquarimicins A-C (1-3), novel natural products with intriguing tetra- or pentacyclic frameworks, has been achieved. The synthesis features an extensive investigation of the biosynthesis-based intramolecular Diels-Alder (IMDA) reactions of (E,Z,E)-1,6,8-nonatrienes. Considering possible biosynthetic sequences, four types of substrates were synthesized, and their IMDA reactions were examined. From one of the four substrates, the total synthesis was achieved via a transannular Diels-Alder reaction, which led to the stereoselective construction of the unique molecular framework. The convergent and efficient synthetic pathway afforded (+)-1 in 27 linear steps with 4.3% and 9.9% overall yields from readily available ethyl (2E,4S)-4,5 (isopropylidene)dioxy-2-pentenoate (22) and (R)-epichlorohydrin (30), respectively. Furthermore, efficient syntheses of 2, 3, and the 9-epi cochleamycins A (57) and B (58) were accomplished. Additionally, the present work established the absolute stereochemistry of macquarimicins and revised the C(2)- C(3) geometry of 1. PMID- 15355108 TI - Mechanistic characterization of aerobic alcohol oxidation catalyzed by Pd(OAc)(2)/pyridine including identification of the catalyst resting state and the origin of nonlinear [catalyst] dependence. AB - The Pd(OAc)(2)/pyridine catalyst system is one of the most convenient and versatile catalyst systems for selective aerobic oxidation of organic substrates. This report describes the catalytic mechanism of Pd(OAc)(2)/pyridine-mediated oxidation of benzyl alcohol, which has been studied by gas-uptake kinetic methods and (1)H NMR spectroscopy. The data reveal that turnover-limiting substrate oxidation by palladium(II) proceeds by a four-step pathway involving (1) formation of an adduct between the alcohol substrate and the square-planar palladium(II) complex, (2) proton-coupled ligand substitution to generate a palladium-alkoxide species, (3) reversible dissociation of pyridine from palladium(II) to create a three-coordinate intermediate, and (4) irreversible beta-hydride elimination to produce benzaldehyde. The catalyst resting state, characterized by (1)H NMR spectroscopy, consists of an equilibrium mixture of (py)(2)Pd(OAc)(2), 1, and the alcohol adduct of this complex, 1xRCH(2)OH. These in situ spectroscopic data provide direct support for the mechanism proposed from kinetic studies. The catalyst displays higher turnover frequency at lower catalyst loading, as revealed by a nonlinear dependence of the rate on [catalyst]. This phenomenon arises from a competition between forward and reverse reaction steps that exhibit unimolecular and bimolecular dependences on [catalyst]. Finally, overoxidation of benzyl alcohol to benzoic acid, even at low levels, contributes to catalyst deactivation by formation of a less active palladium benzoate complex. PMID- 15355109 TI - Zirconium-catalyzed enantioselective [3+2] cycloaddition of hydrazones to olefins leading to optically active pyrazolidine, pyrazoline, and 1,3-diamine derivatives. AB - Asymmetric [3+2] cycloaddition of hydrazones to external olefins has been successfully conducted in high yields with high enantioselectivities using a chiral zirconium catalyst. These reactions open ways to synthetically and biologically important pyrazoline, pyrazolidine, and 1,3-diamine derivatives. Further, several experiments suggested that the reactions proceeded via concerted pathways. PMID- 15355110 TI - Electron impact ionization in helium nanodroplets: controlling fragmentation by active cooling of molecular ions. AB - Reported here is a study of the effects of liquid helium cooling on the fragmentation of ions formed by electron impact mass ionization. The molecules of interest are picked up by the helium nanodroplets as they pass through a low pressure oven. Electron impact ionization of a helium atom in the droplet is followed by resonant charge transfer to neighboring helium atoms. When the charge is transferred to the target molecule, the difference in the ionization potentials between helium and the molecule results in the formation of a vibrationally hot ion. In isolation, the hot parent ion would undergo subsequent fragmentation. On the other hand, if the cooling due to the helium is fast enough, the parent ion will be actively cooled before fragmentation occurs. The target molecule used in the present study is triphenylmethanol (TPM), an important species in synthetic chemistry, used to sterically protect hydroxyl groups. Threshold PhotoElectron PhotoIon COincidence (TPEPICO) experiments are also reported for gas-phase TPM to help quantify the ion energetics resulting from the cooling effects of the helium droplets. PMID- 15355111 TI - Experimental and theoretical analysis of the photochemistry and thermal reactivity of ethyl diazomalonate and its diazirino isomer. The role of molecular geometry in the decomposition of diazocarbonyl compounds. AB - The photochemical or thermal decomposition of ethyl diazomalonate (1) or ethyl 3,3-diazirinedicarboxylate in methanol solutions yields the O-H insertion product 6, while products of the Wolff rearrangement were not detected in both cases. The analysis of temperature-dependent (13)C NMR spectra and the results of DFT B3LYP/6-311+G(3df,2p) and MP2/aug-cc-pVTZ//B3LYP/6-311+G(3df,2p) calculations allow us to conclude that diazodiester 1 predominantly exists in the Z,Z conformation. In contrast, photolysis of the cyclic isopropylidene diazomalonate (3), which also has a Z,Z-configuration of the diazodicarbonyl moiety, results in a clean Wolff rearrangement. These observations allow us to conclude that the direction of the photodecomposition of diazomalonates is not controlled by the ground-state conformation. The quantum-mechanical analysis of the potential energy surfaces for the dediazotization of 1 and 3 suggests that the formation of a carbene as a discrete intermediate is controlled by the ability of the latter to adopt a conformation in which carbonyl groups are almost orthogonal to the carbene plane. The outcome of the photolysis of ethyl diazomalonate depends on the wavelength of irradiation. Irradiation with 254 nm light results in the loss of nitrogen and the formation of dicarboethoxycarbene (5, Phi(254) = 0.31), while at longer wavelengths, diazirine 2 becomes an important byproduct (Phi(350) = 0.09). This observation suggests that the formation of carbene 5 and isomerization to diazirine proceed from different electronically excited states of ethyl diazomalonate. PMID- 15355112 TI - Thermodynamics of diamond nucleation on the nanoscale. AB - To have a clear insight into the diamond nucleation upon the hydrothermal synthesis and the reduction of carbide (HSRC), we performed the thermodynamic approach on the nanoscale to elucidate the diamond nucleation taking place in HSRC supercritical-fluid systems taking into account the capillary effect of the nanosized curvature of the diamond critical nuclei, based on the carbon thermodynamic equilibrium phase diagram. These theoretical analyses showed that the nanosize-induced interior pressure of diamond nuclei could drive the metastable phase region of the diamond nucleation in HSRC into the new stable phase region of diamond in the carbon phase diagram. Accordingly, the diamond nucleation is preferable to the graphite phase formation in the competing growth between diamond and graphite upon HSRC. Meanwhile, we predicted that 400 MPa should be the threshold pressure for the diamond synthesis by HSRC in the metastable phase region of diamond, based on the proposed thermodynamic nucleation on the nanoscale. PMID- 15355113 TI - Superhelices of poly[2-(acetoacetoxy)ethyl methacrylate]. AB - Poly[2-(acetoacetoxy)ethyl methacrylate] (PAEMA) homopolymers were found to self assemble into hierarchical superstructures, that is, double-stranded helical tubes of either screw sense (scanning force microscopy). Both the diameter and the pitch of the superhelices are approximately 12 nm, and their length is 200 500 nm. It is proposed that PAEMA chains first organize into ribbons, the width of which determines the pitch of the helix, and then coil up into the helical superstructure. The formation of these structures is driven by the establishment of hydrogen-bridging interactions between adjacent acetoacetoxy groups (NMR and dielectric relaxation spectroscopy) and compensation of dipole moments. PMID- 15355114 TI - Controlled self-assembly of hexa-peri-hexabenzocoronenes in solution. AB - Disc-shaped hexa-peri-hexabenzocoronenes (HBCs) peripherally substituted by flexible dodecyl chains (molecule 1) or rigid polyphenylene dendrons (molecules 2a,b and 3) were efficiently synthesized. Steric hindrance arising from the substituents, from less hindered dodecyl to bulky dendrons, was utilized to program the self-assembly of the HBC cores in solution. The high tendency of the hexadodecyl-substituted HBC 1 to aggregate was determined by concentration and temperature-dependent (1)H NMR spectroscopic measurements and nonlinear least squares analysis of the experimental data. The rigid dendrons in molecule2a suppress the pi--pi interactions of the HBC cores to a certain extent, and a slow (with respect to the NMR time scale) monomer--dimer equilibrium is observed. This unique equilibrium was further controlled by temperature, concentration, and solvent to afford discrete monomeric or dimeric species. Further structural modifications such as the replacement of dodecyl groups in 2a with hydrogen atoms resulted in a stable dimer structure in 2b due to diminished steric hindrance, as supported by quantum chemical calculations. "Moving" the dendron arms closer to the HBC core gives molecule 3, which exists only as a nonaggregated monomer. UV vis absorption and fluorescence spectra of these discrete species revealed obvious differences in their electronic and optoelectronic properties which can be explained by the existence or absence of pi--pi interactions. PMID- 15355115 TI - Quantum dots tailored with poly(para-phenylene vinylene). AB - In polymernanoparticle composites, uniform dispersion of the nanoparticles carries advantages over cases where nanoparticle aggregation dominates. Such dispersion has been particularly difficult to obtain in the case of composites prepared from nanoparticles and conjugated polymers. Here, we show that cadmium selenide nanocrystals, or quantum dots, can be integrated into thin films of poly(para-phenylene vinylene) (PPV) without aggregation. The two key departures from previous studies of quantum-dot/electronic polymer composites are (1) the synthesis of high-quality quantum dots directly in novel, functional ligands, thus eliminating the need for ligand exchange, and (2) polymerization chemistry that grafts PPV to the quantum dot surface. Solid-state photoluminescence spectra of composite materials prepared by these novel techniques reveal the critical importance of the quantum dot-polymer interface that will enable new investigations in nanoparticle-based light-emitting devices. PMID- 15355117 TI - Short lexitropsin that recognizes the DNA minor groove at 5'-ACTAGT-3': understanding the role of isopropyl-thiazole. AB - Isopropyl-thiazole ((iPr)Th) represents a new addition to the building blocks of nucleic acid minor groove-binding molecules. The DNA decamer duplex d(CGACTAGTCG)(2) is bound by a short lexitropsin of sequence formyl-PyPy(iPr)Th Dp (where Py represents N-methyl pyrrole, (iPr)Th represents thiazole with an isopropyl group attached, and Dp represents dimethylaminopropyl). NMR data indicate ligand binding in the minor groove of DNA to the sequence 5' ACT(5)AG(7)T-3' at a 2:1 ratio of ligand to DNA duplex. Ligand binding, assisted by the enhanced hydrophobicity of the (iPr)Th group, occurs in a head-to-tail fashion, the formyl headgroups being located toward the 5'-ends of the DNA sequence. Sequence reading is augmented through hydrogen bond formation between the exocyclic amine protons of G(7) and the (iPr)Th nitrogen, which lies on the minor groove floor. The B(I)/B(II) DNA backbone equilibrium is altered at the T(5) 3'-phosphate position to accommodate a B(II) configuration. The ligands bind in a staggered mode with respect to one another creating a six base pair DNA reading frame. The introduction of a new DNA sequence-reading element into the recognition jigsaw, combined with an extended reading frame for a small lexitropsin with enhanced hydrophobicity, holds great promise in the development of new, potentially commercially viable drug lead candidates for gene targeting. PMID- 15355118 TI - Probing the location and distribution of paramagnetic centers in alkali metal loaded zeolites through (7)Li MAS NMR. AB - The nature and surroundings of lithium cations in lithium-exchanged X and A zeolites following loading with the alkali metals Na, K, Rb, and Cs have been studied through (7)Li solid-state NMR spectroscopy. It is demonstrated that the lithium in these zeolites is stable with respect to reduction by the other alkali metals. Even though the lithium cations are not directly involved in chemical interactions with the excess electrons introduced in the doping process, the corresponding (7)Li NMR spectra are extremely sensitive to paramagnetic species that are located inside the zeolite cavities. This sensitivity makes (7)Li NMR a useful probe to study the formation, distribution, and transformation of such species. PMID- 15355116 TI - The migrastatin family: discovery of potent cell migration inhibitors by chemical synthesis. AB - The first asymmetric total synthesis of (+)-migrastatin (1), a macrolide natural product with anti-metastatic properties, has been accomplished. Our concise and flexible approach utilized a Lewis acid-catalyzed diene aldehyde condensation (LACDAC) to install the three contiguous stereocenters and the trisubstituted (Z) alkene of migrastatin (2 + 3 --> 21). Construction of the two remaining stereocenters and incorporation of the glutarimide-containing side chain was achieved by an anti-selective aldol addition of propionyl oxazolidinone 28 to angelic aldehyde 27, followed by a Horner-Wadsworth-Emmons (HWE) coupling of 32 with glutarimide aldehyde 5. Finally, the assembly of the macrocycle was realized by a highly (E)-selective ring-closing metathesis (35 --> 37). Utilizing the power of diverted total synthesis (DTS), a series of otherwise inaccessible analogues was prepared and evaluated for their potential as tumor cell migration inhibitors in several in vitro assays. These studies revealed a dramatic increase in activity when the natural motif was considerably simplified, presenting macrolactones 45 and 48, as well as macrolactam 55, macroketone 60, and CF(3) alcohol 71 as promising anti-metastatic agents. PMID- 15355119 TI - Electronic properties of mononuclear, dinuclear, and polynuclear cobaltacarboranes: electrochemical and spectroelectrochemical studies. AB - Electronic interactions and metal-metal communication in a wide range of cobaltacarborane-hydrocarbon complexes containing one to six metal centers, and exhibiting a variety of modes of inter-cage connectivity and molecular architectures, have been investigated via cyclic voltammetry, controlled potential coulometry, and UV-visible spectroelectrochemistry. The properties of mixed-valent Co(III)/Co(IV) and Co(II)/Co(III) species that are generated on oxidation or reduction of dinuclear and polynuclear Co(III) complexes were examined and classified as Robin-Day Class I (localized), Class II (partially delocalized), or Class III (fully delocalized) systems. The extent of metal-metal communication between metallacarborane cage units is strongly influenced by the type of intercage connection (e.g., cage B-B or Cp-Cp); the vertexes involved (equatorial vs apical); the nature of the linking unit, if any; and the presence of substituents on the carborane cages. In multi-tripledecker complexes where three CpCo(C(2)B(3)H(4))CoCp units are linked through a central triethynyl benzene connector, the data suggest that Co-Co electronic communication is extensive (Class III) within individual sandwich units while intersandwich delocalization is weak or absent. An extended Huckel study of CpCoC(2)B(4)H(6) double-decker and CpCo(C(2)B(3)H(5))CoCp triple-decker sandwich model complexes shows significant differences in the orbital contributions involved in the HOMO and LUMO of the former vs the latter type. The calculations afford additional insight into the electronic structures and properties of these systems as elucidated by the experimental studies. PMID- 15355120 TI - A homoleptic molybdenum(IV) enolate complex: synthesis, molecular and electronic structure, and NCN group transfer to form a terminal cyanoimide of molybdenum(VI). AB - A monomeric molybdenum(IV) tetrakis enolate complex Mo(OC[Ad]Mes)(4), 1, where Ad = 2-adamantylidene and Mes = 2,4,6-Me(3)C(6)H(2), has been synthesized and characterized structurally by X-ray diffraction, chemically through NCN group transfer reactivity, and computationally to investigate the origins of the observed structure that is intermediate between tetrahedral and square planar. No prior examples of Mo(OR)(4) have been structurally characterized despite having been the subject of both experimental and theoretical interest. Complex 1 has a singlet ground state and thus a metal-based lone pair of electrons. The latter has been visualized with the aid of the electron localization function (ELF) and appears as a two-bladed propeller with D(2)(d)() symmetry. Complex 1 makes a simple 1:1 adduct with t-BuNC that is trigonal bipyramidal with an axial isocyanide as demonstrated by X-ray crystallography. This trigonal bipyramidal 1:1 adduct has a triplet ground state and provides a model for the way in which 1 interacts with NCN group donor dbabhCN prior to NCN group transfer to form the terminal cyanoimide complex 1-NCN. The calculated Mo-N bond dissociation enthalpy for 1-NCN is 104 kcal mol(-1), 30 kcal mol(-1) greater than that for the corresponding dissociation of NCN from cyanophosphiniminato NCNPMe(3). PMID- 15355121 TI - Understanding the role of sodium during adsorption: a force field for alkanes in sodium-exchanged faujasites. AB - We have developed a united atom force field able to accurately describe the adsorption properties of linear alkanes in the sodium form of FAU-type zeolites. This force field successfully reproduces experimental adsorption properties of n alkanes over a wide range of sodium cation densities, temperatures, and pressures. The force field reproduces the sodium positions in dehydrated FAU-type zeolites known from crystallography, and it predicts how the sodium cations redistribute when n-alkanes adsorb. The cations in the sodalite cages are significantly more sensitive to the n-alkane loading than those in the supercages. We provide a simple expression that adequately describes the n-alkane Henry coefficient and adsorption enthalpy as a function of sodium density and temperature at low coverage. This expression affords an adequate substitute for complex configurational-bias Monte Carlo simulations. The applicability of the force field is by no means limited to low pressure and pure adsorbates, for it also successfully reproduces the adsorption from binary mixtures at high pressure. PMID- 15355122 TI - Charge redistribution on electronic excitation. Dipole moments of cis- and trans 3-aminophenol in their S(0) and S(1) electronic states. AB - Rotationally resolved electronic spectroscopy in the gas phase, in the absence and presence of an applied electric field, has been used to distinguish the two conformers of 3-aminophenol (3AP) on the basis of differences in their electric dipole moments. cis-3AP has micro = 2.3 D, and trans-3AP has micro = 0.7 D, in their ground electronic states. The two observed values are approximately equal to those expected on the basis of bond dipole additivity rules. However, these rules fail to predict the large change in both the magnitude and the orientation of micro when the two conformers of 3AP absorb light. cis-3AP has micro = 3.3 D, and trans-3AP has micro = 1.7 D, in their excited S(1) electronic states; the angles of orientation of micro with respect to the a inertial axis change by 13 degrees and 38 degrees, respectively. This effect is attributed to (1)L(b)/(1)L(a) state mixing in the S(1) state. PMID- 15355123 TI - The photoinduced triplet of flavins and its protonation states. AB - The photogenerated triplet states of riboflavin and flavin mononucleotide (FMN) have been examined by time-resolved electron paramagnetic resonance (EPR) spectroscopy at low temperature (T = 80 K). Because of the high time resolution of the utilized EPR instrumentation, the triplets are for the first time observed in the nonequilibrated electron-spin polarized state and not in their equilibrated forms with the population of the triplet sublevels governed by Boltzmann distribution. The electron-spin polarization pattern directly reflects the anisotropy of the intersystem crossing from the excited singlet-state precursor. Spectral analysis of the resulting enhanced absorptive and emissive EPR signals yields the zero-field splitting parameters, |D| and |E|, and the zero field populations of the triplet at high accuracy. These parameters are sensitive probes for the protonation state of the flavin's isoalloxazine ring, as becomes evident by a comparison of the spectra recorded at different pH values of the solvent. The three protonation states of the flavins can furthermore be distinguished by the kinetics of the transient EPR signals, which are dominated by spin-lattice relaxation. The fastest decays are observed for the protonated FMN and riboflavin triplets, followed by the deprotonated flavin triplets. Slow decays are measured for the triplet states of neutral FMN and riboflavin. Because proton transfer is found to be slow on the time scale of spin-polarized triplet detection by transient EPR, the pH-dependent spin-relaxation and zero-field splitting parameters offer a novel approach to probe the protonation state of flavins in their singlet ground state through the characterization of their triplet-state properties. PMID- 15355124 TI - Pharmacokinetic interactions of topiramate. AB - Topiramate is a new antiepileptic drug (AED) that has been approved worldwide (in more than 80 countries) for the treatment of various kinds of epilepsy. It is currently being evaluated for its effect in various neurological and psychiatric disorders. The pharmacokinetics of topiramate are characterised by linear pharmacokinetics over the dose range 100-800 mg, low oral clearance (22-36 mL/min), which, in monotherapy, is predominantly through renal excretion (renal clearance 10-20 mL/min), and a long half-life (19-25 hours), which is reduced when coadministered with inducing AEDs such as phenytoin, phenobarbital and carbamazepine. The absolute bioavailability, or oral availability, of topiramate is 81-95% and is not affected by food. Although topiramate is not extensively metabolised when administered in monotherapy (fraction metabolised approximately 20%), its metabolism is induced during polytherapy with carbamazepine and phenytoin, and, consequently, its fraction metabolised increases. During concomitant treatment with topiramate and carbamazepine or phenytoin, the (oral) clearance of topiramate increases 2-fold and its half-life becomes shorter by approximately 50%, which may require topiramate dosage adjustment when phenytoin or carbamazepine therapy is added or discontinued. From a pharmacokinetic standpoint, topiramate is a unique example of a drug that, because of its major renal elimination component, is not subject to drug interaction due to enzyme inhibition, but nevertheless is susceptible to clinically relevant drug interactions due to induction of its metabolism. Unlike old AEDs such as phenytoin and carbamazepine, topiramate is a mild inducer and, currently, the only interaction observed as a result of induction by topiramate is that with ethinylestradiol. Topiramate only increases the oral clearance of ethinylestradiol in an oral contraceptive at high dosages (>200 mg/day). Because of this dose-dependency, possible interactions between topiramate and oral contraceptives should be assessed according to the topiramate dosage utilised. This paper provides a critical review of the pharmacokinetic interactions of topiramate with old and new AEDs, an oral contraceptive, and the CNS-active drugs lithium, haloperidol, amitriptyline, risperidone, sumatriptan, propranolol and dihydroergotamine. At a daily dosage of 200 mg, topiramate exhibited no or little (with lithium, propranolol and the amitriptyline metabolite nortriptyline) pharmacokinetic interactions with these drugs. The results of many of these drug interaction studies with topiramate have not been published before, and are presented and discussed for the first time in this article. PMID- 15355125 TI - Clinical pharmacokinetics and pharmacodynamics of inhaled insulin. AB - The benefits of intensive insulin therapy in the prevention of complications in patients with diabetes mellitus are now well established. However, the current methods of insulin administration fall well short of the ideal. Consequently, alternative routes of insulin administration have been investigated. The pulmonary route has received the most attention, helped by advances in inhaler devices and insulin formulation technology. As a result, several insulin inhalation systems are at varying stages of development, with one already filed for marketing approval in Europe. Knowledge of the pharmacokinetic and pharmacodynamic characteristics of the various inhaled insulin formulations will help to determine their positioning in current and evolving diabetes treatment strategies. For instance, a rapid onset and short duration of action would be desirable for use in postprandial glucose control. Pharmacokinetic studies with inhaled insulin reveal that serum insulin concentrations peak earlier and decay more rapidly following inhalation compared with subcutaneously administered regular insulin, and pharmacodynamic studies measuring glucose infusion rate under euglycaemic glucose clamp show corresponding rapid changes in glucose control. Furthermore, intrapatient variability in the pharmacokinetics and pharmacodynamics of inhaled insulin is low; variability is similar to (or perhaps less than) that seen when insulin is administered subcutaneously. Estimates of the bioavailability and bioefficacy achievable with the current inhalation systems are typically in the region of 10% of that experienced with subcutaneously administered insulin. Most of the losses are in the device, mouth and throat, with approximately 30-50% of the insulin deposited in the lungs being absorbed. Clinical experience to date indicates that inhaled insulin has the potential to be an effective treatment in patients with diabetes, and that it may have particular utility in the treatment of postprandial hyperglycaemia. PMID- 15355126 TI - Pharmacokinetics of budesonide (Entocort EC) capsules for Crohn's disease. AB - This overview summarises available pharmacokinetic data on budesonide capsules (Entocort EC), approved for the treatment of mild-to-moderate active Crohn's disease involving the ileum and/or ascending colon and for prolongation of symptom control. Budesonide is a locally-acting glucocorticosteroid with an extensive, primarily hepatic, metabolism after oral administration. It is rapidly absorbed and biotransformed by cytochrome P450 (CYP) 3A to metabolites with negligible glucocorticoid activity. Entocort EC, a pH- and time-dependent oral formulation of budesonide, was developed to optimise drug delivery to the ileum and throughout the colon. Pharmaco-scintigraphic studies have confirmed that the Entocort EC formulation delays budesonide absorption and prolongs the rate of elimination but maintains complete absorption. This improves the delivery of budesonide to the intestinal lumen relative to a plain formulation. A low systemic availability of 9-21% indicates extensive first-pass elimination. Food appears to have little impact on the absorption of budesonide from Entocort EC capsules and the pharmacokinetics are dose-proportional between 3 and 15 mg. On average, systemic availability was 2.5-fold higher in patients with cirrhosis compared with healthy controls; however, mild liver impairment had little effect on systemic exposure. Pharmacokinetics appear unaffected by gender and age, although this has not been tested in younger children. Renal impairment is not expected to have an impact on the kinetics of Entocort EC. Budesonide is unlikely to inhibit the metabolism of other drugs, including CYP3A4 substrates, mainly because of the very low plasma concentrations obtained with the compound even after high doses of Entocort trade mark EC capsules. Strong CYP3A4 inhibitors, such as ketoconazole, will inhibit the metabolism of budesonide, resulting in several-fold increases in the area under the concentration-time curve of budesonide. Also, grapefruit juice intake may increase systemic availability of budesonide, probably by inhibition of intestinal CYP3A4 activity. Unlike prednisolone, oral contraceptives do not alter plasma budesonide concentrations. An increased pH obtained by gastric acid inhibitory drugs, such as omeprazole, does not affect the pharmacokinetics of budesonide. In summary, budesonide capsules (Entocort EC) possess many pharmacological features that make the formulation well adapted for a targeted treatment of inflammatory disorders, such as Crohn's disease involving the ileum and ascending colon. PMID- 15355127 TI - Effect of mycophenolate mofetil on the pharmacokinetics of antiretroviral drugs and on intracellular nucleoside triphosphate pools. AB - OBJECTIVE: To study the effect of mycophenolate mofetil therapy on the pharmacokinetic parameters of a number of antiretroviral drugs, on intracellular pools of deoxycytidine triphosphate (dCTP) and deoxyguanosine triphosphate (dGTP), and on intracellular concentrations of the triphosphate of lamivudine (3TCTP). DESIGN: Randomised pharmacokinetic study. PARTICIPANTS: Nineteen HIV-1 infected patients. METHODS: Antiretroviral-naive men starting treatment with didanosine 400 mg once daily, lamivudine 150 mg twice daily, abacavir 300 mg twice daily, indinavir 800 mg twice daily, ritonavir 100 mg twice daily and nevirapine 200 mg twice daily were randomised to a group with or without mycophenolate mofetil 500 mg twice daily. After 8 weeks of therapy, the plasma pharmacokinetic profiles of mycophenolic acid (the active metabolite of mycophenolate mofetil), abacavir, indinavir and nevirapine, and triphosphate concentrations (dCTP, dGTP and 3TCTP) in peripheral blood mononuclear cells, were determined. RESULTS: Nine of the 19 patients received mycophenolate mofetil. There was no difference in plasma clearance of indinavir or abacavir between the two groups. The clearance of nevirapine was higher in patients using mycophenolate mofetil (p = 0.04). In 12 patients, of whom five also received mycophenolate mofetil, intracellular triphosphates were measured. There was no significant difference in intracellular dCTP, dGTP or 3TCTP concentrations between the two groups. CONCLUSION: In this small cohort of patients, mycophenolate mofetil therapy reduced the plasma concentration of nevirapine but had no effect on plasma concentrations of indinavir and abacavir. There were no consistent effects of mycophenolic acid on the intracellular concentrations of dCTP, dGTP or 3TCTP. PMID- 15355128 TI - Explaining purportedly irrational behavior by modeling skepticism in task parameters: an example examining confidence in forced-choice tasks. AB - Many purported demonstrations of irrational behavior rely on the assumption that participants believe key task parameters that are merely asserted by experimenters. For example, previous researchers have found that participants who first reported confidence in items presented in a yes-no format did not change confidence to the degree prescribed by the normative model when those same items were later presented in a forced-choice format. A crucial assumption, however, was that participants fully believed the assertion that the forced-choice items were mutually exclusive and exhaustive. In this article, the authors derive and test a new normative model in which it is not assumed that participants fully believe the assertion. Two visual identification experiments show that the new normative model provides a compelling account of participants' confidence reports. PMID- 15355129 TI - The "saw-it-all-along" effect: demonstrations of visual hindsight bias. AB - The authors address whether a hindsight bias exists for visual perception tasks. In 3 experiments, participants identified degraded celebrity faces as they resolved to full clarity (Phase 1). Following Phase 1, participants either recalled the level of blur present at the time of Phase 1 identification or predicted the level of blur at which a peer would make an accurate identification. In all experiments, participants overestimated identification performance of naive observers. Visual hindsight bias was greater for more familiar faces--those shown in both phases of the experiment--and was not reduced following instructions to participants to avoid the bias. The authors propose a fluency-misattribution theory to account for the bias and discuss implications for medical malpractice litigation and eyewitness testimony. PMID- 15355130 TI - Three turtles in danger: spontaneous construction of causally relevant spatial situation models. AB - In 4 experiments, the author explored the spontaneous construction of spatial situation models during discourse comprehension by using the sentence-recognition paradigm of J. D. Bransford, J. R. Barclay, and J. J. Franks (1972). In Experiment 1, signaling causal relevance of spatial relations was a necessary precondition for replicating their original finding of spontaneously constructed spatial representations. Causal relevance was ensured in the subsequent experiments by a judgment task indirectly demanding the evaluation of described spatial relations with regard to causal relevance. Participants spontaneously constructed spatial situation models of text presented auditorily or visually. Effects of spontaneous construction were more reliable when encoding was easier. The results suggest a revised interpretation of J. D. Bransford et al.'s study and corroborate recent evidence showing that relevant spatial information in texts is reliably represented. PMID- 15355131 TI - Interactive dimensions in the construction of mental representations for text. AB - To detail the structure and format of memory for texts, researchers have examined whether readers monitor separate text dimensions for space, time, and characters. The authors proposed that the interactivity between these individual dimensions may be as critical to the construction of complex mental models as the discrete dimensions themselves. In the present experiments, participants read stories in which characters were described as traveling from a start to a final location. During movement between locations, characters engaged in activities that could take either a long or short amount of time to complete. Results indicate that accessibility for the spatial locations was a function of the passage of time. The authors interpret this as evidence that the interactive nature of text dimensions affects the structure of representations in memory. PMID- 15355132 TI - Effects of model-based and memory-based processing on speed and accuracy of grammar string generation. AB - Learners are able to use 2 different types of knowledge to perform a skill. One type is a conscious mental model, and the other is based on memories of instances. The authors conducted 3 experiments that manipulated training conditions designed to affect the availability of 1 or both types of knowledge about an artificial grammar. Participants were tested for both speed and accuracy of their ability to generate letter sequences. Results indicate that model-based training leads to slow accurate responding. Memory-based training leads to fast, less accurate responding and highest achievement when perfect accuracy was not required. Evidence supports participants' preference for using the memory-based mode when exposed to both types of training. Finally, the accuracy contributed by model-based training declined over a retention interval. PMID- 15355133 TI - Effects of additional tasks on language perception: an event-related brain potential investigation. AB - The authors investigated effects of task and overlapping processing load on semantic processing. In 3 experiments the brain potential component N400 was elicited by synonymous and nonsynonymous spoken noun pairs that were to be classified according to semantic relatedness. The time course of the N400 component to the nouns was delayed, and its amplitude was reduced when additional tasks had to be performed. The delay increased with temporal overlap and was more pronounced for language-related than for spatial additional tasks. Delays of N400 were also caused by a compatibility manipulation in the additional tasks. Theoretical accounts of the observed interference of overlapping tasks with language perception in terms of attentional shifts were discarded. Explanations in terms of resource sharing and single channel processing are discussed. PMID- 15355134 TI - Multidimensional rule, unidimensional rule, and similarity strategies in categorization: event-related brain potential correlates. AB - Forty participants assigned artificial creatures to categories after explicit rule instruction or feedback alone. Stimuli were typical and atypical exemplars of 2 categories with independent prototypes, conflicting exemplars sharing features of both categories, and "Others" with only 1 or 2 features of the well defined categories. Ten feedback-only participants spontaneously adopted a unidimensional rule; 10 used a multidimensional similarity strategy. Event related potentials (ERPs) recorded during the transfer phase showed a commonality between multidimensional rule and similarity strategies in late frontal brain activity that differentiated both from unidimensional rule use. Multidimensional rule users alone showed an earlier prefrontal ERP effect that may reflect inhibition of responses based on similarity. The authors also discuss the role of declarative memory for features and exemplars. PMID- 15355135 TI - Knowledge partitioning in categorization: constraints on exemplar models. AB - The authors present 2 experiments that establish the presence of knowledge partitioning in perceptual categorization. Many participants learned to rely on a context cue, which did not predict category membership but identified partial boundaries, to gate independent partial categorization strategies. When participants partitioned their knowledge, a strategy used in 1 context was unaffected by knowledge demonstrably present in other contexts. An exemplar model, attentional learning covering map, was shown to be unable to accommodate knowledge partitioning. Instead, a mixture-of-experts model, attention to rules and instances in a unified model (ATRIUM), could handle the results. The success of ATRIUM resulted from its assumption that people memorize not only exemplars but also the way in which they are to be classified. PMID- 15355136 TI - Modeling the effects of prior knowledge on learning incongruent features of category members. AB - The authors conducted 3 experiments addressing the issue of how observations and multiple sources of prior knowledge are put together in category learning. In Experiments 1 and 2, learning was faster for critical features, which were predictable on the basis of prior knowledge, than for filler features, and this advantage increased as more observations were made. In addition, learning was fastest for incongruent features that could only be predicted using knowledge from other domains. In Experiment 3, presenting contradictory features that violated prior knowledge led to rote learning rather than use of prior knowledge. The results were simulated with the Baywatch model, which addresses how observations of category members lead to recruitment and selection of sources of prior knowledge. PMID- 15355137 TI - The importance of being nonalignable: a critical test of the structural alignment theory of similarity. AB - The structural alignment theory of similarity distinguishes 2 types of difference that may occur between stimuli: Alignable differences are those related to a commonality, whereas nonalignable differences are not related to a commonality. Alignment theory predicts that alignable differences should be more heavily weighted than nonalignable differences in similarity judgment. Experiments 1 and 2 demonstrate that, contrary to this prediction, nonalignable differences exerted a greater impact than alignable differences in similarity and difference judgments of geometric stimuli. Experiment 3 revealed that the relative weight accorded a given difference was also affected by contextual constraints. Thus, although the experiments supported the validity of the distinction between alignable and nonalignable differences, results were discordant with the specific prediction of structural alignment theory. PMID- 15355138 TI - Retroactive effects of irrelevant speech on serial recall from short-term memory. AB - The authors report 5 serial-recall experiments. In 4 of the 5 experiments, they show that irrelevant sound (IS) has a retroactive effect on material already in memory. In Experiment 1, IS presented during a filled retention interval had a reliable effect on list recall. Four further experiments, 3 of which used retroactive IS, showed that IS continued to-have an effect on recall following a long, filled retention interval. Articulatory suppression during visual input was found to abolish the long-lasting, retroactive effect of IS, supporting the idea that IS affects the phonological-loop component of short-term memory. IS also, therefore, seems to affect a longer term memory system with which the loop interacts. PMID- 15355139 TI - The irrelevant sound phenomenon revisited: what role for working memory capacity? AB - High-span individuals (as measured by the operation span [OSPAN] technique) are less likely than low-span individuals to notice their own names in an unattended auditory stream (A. R. A. Conway, N. Cowan, & M. F. Bunting, 2001). The possibility that OSPAN accounts for individual differences in auditory distraction on an immediate recall test was examined. There was no evidence that high-OSPAN participants were more resistant to the disruption caused by irrelevant speech in serial or in free recall. Low-OSPAN participants did, however, make more semantically related intrusion errors from the irrelevant sound stream in a free recall test (Experiment 4). Results suggest that OSPAN mediates semantic components of auditory distraction dissociable from other aspects of the irrelevant sound effect. PMID- 15355140 TI - Distant melodies: statistical learning of nonadjacent dependencies in tone sequences. AB - Human listeners can keep track of statistical regularities among temporally adjacent elements in both speech and musical streams. However, for speech streams, when statistical regularities occur among nonadjacent elements, only certain types of patterns are acquired. Here, using musical tone sequences, the authors investigate nonadjacent learning. When the elements were all similar in pitch range and timbre, learners acquired moderate regularities among adjacent tones but did not acquire highly consistent regularities among nonadjacent tones. However, when elements differed in pitch range or timbre, learners acquired statistical regularities among the similar, but temporally nonadjacent, elements. Finally, with a moderate grouping cue, both adjacent and nonadjacent statistics were learned, indicating that statistical learning is governed not only by temporal adjacency but also by Gestalt principles of similarity. PMID- 15355141 TI - Implicit learning of musical timbre sequences: statistical regularities confronted with acoustical (dis)similarities. AB - The present study investigated the influence of acoustical characteristics on the implicit learning of statistical regularities (transition probabilities) in sequences of musical timbres. The sequences were constructed in such a way that the acoustical dissimilarities between timbres potentially created segmentations that either supported (S1) or contradicted (S2) the statistical regularities or were neutral (S3). In the learning group, participants first listened to the continuous timbre sequence and then had to distinguish statistical units from new units. In comparison to a control group without the exposition phase, no interaction between sequence type and amount of learning was observed: Performance increased by the same amount for the three sequences. In addition, performance reflected an overall preference for acoustically similar timbre units. The present outcome extends previous data from the domain of implicit learning to complex nonverbal auditory material. It further suggests that listeners become sensitive to statistical regularities despite acoustical characteristics in the material that potentially affect grouping. PMID- 15355142 TI - A rational look at the emotional stroop phenomenon: a generic slowdown, not a stroop effect. AB - The role of Stroop processes in the emotional Stroop effect was subjected to a conceptual scrutiny augmented by a series of experiments entailing reading or lexical decision as well as color naming. The analysis showed that the Stroop effect is not defined in the emotional Stroop task. The experiments showed that reading, lexical decision, and color naming all are slower with emotional words and that this delay is immune to task-irrelevant variation and to changes in the relative salience of the words and the colors. The delay was absent when emotional and neutral words appeared in a single block. A threat-driven generic slowdown is implicated, not a selective attention mechanism associated with the classic Stroop effect. PMID- 15355143 TI - Load theory of selective attention and cognitive control. AB - A load theory of attention in which distractor rejection depends on the level and type of load involved in current processing was tested. A series of experiments demonstrates that whereas high perceptual load reduces distractor interference, working memory load or dual-task coordination load increases distractor interference. These findings suggest 2 selective attention mechanisms: a perceptual selection mechanism serving to reduce distractor perception in situations of high perceptual load that exhaust perceptual capacity in processing relevant stimuli and a cognitive control mechanism that reduces interference from perceived distractors as long as cognitive control functions are available to maintain current priorities (low cognitive load). This theory resolves the long standing early versus late selection debate and clarifies the role of cognitive control in selective attention. PMID- 15355144 TI - Double dissociations in visual and spatial short-term memory. AB - A visual short-term memory task was more strongly disrupted by visual than spatial interference, and a spatial memory task was simultaneously more strongly disrupted by spatial than visual interference. This double dissociation supports a fractionation of visuospatial short-term memory into separate visual and spatial components. In 6 experiments, this interpretation could be defended against alternative explanations in terms of trade-offs in resource allocation between memory tasks and interference tasks, in terms of an involvement of short term consolidation and long-term memory, in terms of differential phonological loop and central-executive involvement, and in terms of similarity-based interference. PMID- 15355145 TI - The role of bottom-up processing in perceptual categorization by 3- to 4-month old infants: simulations and data. AB - Disentangling bottom-up and top-down processing in adult category learning is notoriously difficult. Studying category learning in infancy provides a simple way of exploring category learning while minimizing the contribution of top-down information. Three- to 4-month-old infants presented with cat or dog images will form a perceptual category representation for cat that excludes dogs and for dog that includes cats. The authors argue that an inclusion relationship in the distribution of features in the images explains the asymmetry. Using computational modeling and behavioral testing, the authors show that the asymmetry can be reversed or removed by using stimulus images that reverse or remove the inclusion relationship. The findings suggest that categorization of nonhuman animal images by young infants is essentially a bottom-up process. PMID- 15355146 TI - Category learning in rhesus monkeys: a study of the Shepard, Hovland, and Jenkins (1961) tasks. AB - In influential research, R. N. Shepard, C. I. Hovland, and H. M. Jenkins (1961) surveyed humans' categorization abilities using tasks based in rules, exclusive or (XOR) relations, and exemplar memorization. Humans' performance was poorly predicted by cue-conditioning or stimulus-generalization theories, causing Shepard et al. to describe it in terms of hypothesis selection and rule application that were possibly supported by verbal mediation. The authors of the current article surveyed monkeys' categorization abilities similarly. Monkeys, like humans, found category tasks with a single relevant dimension the easiest and perceptually chaotic tasks requiring exemplar memorization the most difficult. Monkeys, unlike humans, found tasks based in XOR relations very difficult. The authors discuss the character and basis of the species difference in categorization and consider whether monkeys are the generalization-based cognitive system that humans are not. PMID- 15355147 TI - Having the memory of an elephant: long-term retrieval and the use of analogues in problem solving. AB - The authors report 4 experiments exploring long-term analogical transfer from problem solutions in folk tales participants heard during childhood, many years before encountering the target problems. Substantial culture-specific analogical transfer was found when American and Chinese participants' performance was compared on isomorphs of problems solved in European versus Chinese folk tales. There was evidence of transfer even among participants who did not report being reminded of the source tale while solving the target problem. Comparisons of different versions of a target problem indicated that similarity of solution tool affected accessing, mapping, and executing components of problem solving, whereas similarity of goal object had only a moderate effect on accessing. High school students also evidenced greater transfer than did middle school students. PMID- 15355148 TI - The collider principle in causal reasoning: why the Monty Hall dilemma is so hard. AB - The authors tested the thesis that people find the Monty Hall dilemma (MHD) hard because they fail to understand the implications of its causal structure, a collider structure in which 2 independent causal factors influence a single outcome. In 4 experiments, participants performed better in versions of the MHD involving competition, which emphasizes causality. This manipulation resulted in more correct responses to questions about the process in the MHD and a counterfactual that changed its causal structure. Correct responses to these questions were associated with solving the MHD regardless of condition. In addition, training on the collider principle transferred to a standard version of the MHD. The MHD taps a deeper question: When is knowing about one thing informative about another? PMID- 15355149 TI - This construction needs learned. AB - Four experiments are presented in which adults learned to comprehend a new syntactic construction in their native language. Experiments 1 and 2 demonstrate that adults quickly learn to comprehend the new construction and generalize it to new verbs. Experiment 3 shows that experience with the novel construction affects the processing of a construction already known to the participants and with which the novel construction is temporarily ambiguous. Experiment 4 demonstrates that the influence of a novel construction on the comprehension of familiar constructions is affected by the processing that occurred while the novel construction was learned. These results are discussed in the context of the constraint satisfaction approach to sentence processing and episodic-processing accounts of memory. PMID- 15355150 TI - Structural equation models of latent interactions: evaluation of alternative estimation strategies and indicator construction. AB - Interactions between (multiple indicator) latent variables are rarely used because of implementation complexity and competing strategies. Based on 4 simulation studies, the traditional constrained approach performed more poorly than did 3 new approaches--unconstrained, generalized appended product indicator, and quasi-maximum-likelihood (QML). The authors' new unconstrained approach was easiest to apply. All 4 approaches were relatively unbiased for normally distributed indicators, but the constrained and QML approaches were more biased for nonnormal data; the size and direction of the bias varied with the distribution but not with the sample size. QML had more power, but this advantage was qualified by consistently higher Type I error rates. The authors also compared general strategies for defining product indicators to represent the latent interaction factor. PMID- 15355151 TI - Squeezing interval change from ordinal panel data: latent growth curves with ordinal outcomes. AB - A didactic on latent growth curve modeling for ordinal outcomes is presented. The conceptual aspects of modeling growth with ordinal variables and the notion of threshold invariance are illustrated graphically using a hypothetical example. The ordinal growth model is described in terms of 3 nested models: (a) multivariate normality of the underlying continuous latent variables (yt) and its relationship with the observed ordinal response pattern (Yt), (b) threshold invariance over time, and (c) growth model for the continuous latent variable on a common scale. Algebraic implications of the model restrictions are derived, and practical aspects of fitting ordinal growth models are discussed with the help of an empirical example and Mx script (M. C. Neale, S. M. Boker, G. Xie, & H. H. Maes, 1999). The necessary conditions for the identification of growth models with ordinal data and the methodological implications of the model of threshold invariance are discussed. PMID- 15355152 TI - Structured latent curve models for the study of change in multivariate repeated measures. AB - This article considers a structured latent curve model for multiple repeated measures. In a structured latent curve model, a smooth nonlinear function characterizes the mean response. A first-order Taylor polynomial taken with regard to the mean function defines elements of a restricted factor matrix that may include parameters that enter nonlinearly. Similar to factor scores, random coefficients are combined with the factor matrix to produce individual latent curves that need not follow the same form as the mean curve. Here the associations between change characteristics in multiple repeated measures are studied. A factor analysis model for covariates is included as a means of relating latent covariates to the factors characterizing change in different repeated measures. An example is provided. PMID- 15355153 TI - Analyzing psychopathology items: a case for nonparametric item response theory modeling. AB - The authors discuss the applicability of nonparametric item response theory (IRT) models to the construction and psychometric analysis of personality and psychopathology scales, and they contrast these models with parametric IRT models. They describe the fit of nonparametric IRT to the Depression content scale of the Minnesota Multiphasic Personality Inventory--2 (J. N. Butcher, W. G. Dahlstrom, J. R. Graham, A. Tellegen, & B. Kaemmer, 1989). They also show how nonparametric IRT models can easily be applied and how misleading results from parametric IRT models can be avoided. They recommend the use of nonparametric IRT modeling prior to using parametric logistic models when investigating personality data. PMID- 15355154 TI - Bootstrap standard error and confidence intervals for the correlation corrected for range restriction: a simulation study. AB - The standard Pearson correlation coefficient is a biased estimator of the true population correlation, rho, when the predictor and the criterion are range restricted. To correct the bias, the correlation corrected for range restriction, rc, has been recommended, and a standard formula based on asymptotic results for estimating its standard error is also available. In the present study, the bootstrap standard-error estimate is proposed as an alternative. Monte Carlo simulation studies involving both normal and nonnormal data were conducted to examine the empirical performance of the proposed procedure under different levels of rho, selection ratio, sample size, and truncation types. Results indicated that, with normal data, the bootstrap standard-error estimate is more accurate than the traditional estimate, particularly with small sample size. With nonnormal data, performance of both estimates depends critically on the distribution type. Furthermore, the bootstrap bias-corrected and accelerated interval consistently provided the most accurate coverage probability for rho. PMID- 15355155 TI - Properties of the Hubert-Arabie adjusted Rand index. AB - This article provides an investigation of cluster validation indices that relates 4 of the indices to the L. Hubert and P. Arabie (1985) adjusted Rand index--the cluster validation measure of choice (G. W. Milligan & M. C. Cooper, 1986). It is shown how these other indices can be "roughly" transformed into the same scale as the adjusted Rand index. Furthermore, in-depth explanations are given of why classification rates should not be used in cluster validation research. The article concludes by summarizing several properties of the adjusted Rand index across many conditions and provides a method for testing the significance of observed adjusted Rand indices. PMID- 15355156 TI - Does class size in first grade relate to children's academic and social performance or observed classroom processes? AB - This study evaluated the extent to which first-grade class size predicted child outcomes and observed classroom processes for 651 children (in separate classrooms). Analyses examined observed child-adult ratios and teacher-reported class sizes. Smaller classrooms showed higher quality instructional and emotional support, although children were somewhat less likely to be engaged. Teachers in smaller classes rated typical children in those classes as more socially skilled and as showing less externalizing behavior and reported more closeness toward them. Children in smaller classes performed better on literacy skills. Larger classrooms showed more group activities directed by the teacher, teachers and children interacted more often, and children were more often engaged. Lower class sizes were not of more benefit (or harm) as a function of the child's family income. First-grade class size in the range typical of present-day classrooms in the United States predicts classroom social and instructional processes as well as relative changes in social and literacy outcomes from kindergarten to first grade. PMID- 15355157 TI - Phonemes, rimes, vocabulary, and grammatical skills as foundations of early reading development: evidence from a longitudinal study. AB - The authors present the results of a 2-year longitudinal study of 90 British children beginning at school entry when they were 4 years 9 months old (range = 4 years 2 months to 5 years 2 months). The relationships among early phonological skills, letter knowledge, grammatical skills, and vocabulary knowledge were investigated as predictors of word recognition and reading comprehension. Word recognition skills were consistently predicted by earlier measures of letter knowledge and phoneme sensitivity (but not by vocabulary knowledge, rhyme skills, or grammatical skills). In contrast, reading comprehension was predicted by prior word recognition skills, vocabulary knowledge, and grammatical skills. The results are related to current theories about the role of phonological, grammatical, and vocabulary skills in the development of early reading skills. PMID- 15355158 TI - Early development of scaling ability. AB - The map is a small-scaled version of the space it represents. It has been argued that children have difficulty interpreting maps because they do not understand scale relations. Recent research has shown that even preschoolers can solve problems that involve scaling in one dimension. This study examined whether early scaling ability extends to tasks involving two-dimensional maps and referent spaces of different sizes. Results showed that about 60% of the 4-year-olds and 90% of the 5-year-olds tested used distance information presented on a map to locate an object in a two-dimensional spatial layout. Children had more difficulties in solving mapping tasks with a larger referent space. This decrease in accuracy as a function of space size on the mapping task was greater than would have been expected on the basis of performance on a parallel nonmapping task. The results are discussed in terms of their implications for the mechanisms underlying early scaling ability. PMID- 15355159 TI - The role of classroom norms in contextualizing the relations of children's social behaviors to peer acceptance. AB - This study introduces a social context model within which peer acceptances of prosocial-leadership, aggression, and social withdrawal were examined as functions of the contextual norms of these behaviors. The major postulate of the model is that the extent to which a behavior permeates a social context facilitates peer acceptance of the behavior. Specific hypotheses with respect to classroom and gender as different social contexts were formulated and supported when tested in a sample of 4,650 Chinese middle school students from 82 classes. The discussion emphasizes the theoretical as well as the methodological need for alternative conceptualizations of peer relations that reflect both individual differences and contextual variations. PMID- 15355160 TI - Religion as a resource for positive youth development: religion, social capital, and moral outcomes. AB - Although existing literature demonstrates that developmental benefits are associated with religion for adolescents, little is understood about the dynamics of this relationship. Drawing on social capital theory, this study tested a conceptual model exploring socially embedded religious influences on moral outcomes. A three-dimensional model of social capital demonstrated how social interaction, trust, and shared vision enable social ties associated with religiousness to influence moral behavior. Structural equation modeling was used with data gathered from 735 urban youths to test a proposed model of the effects of religiousness on moral outcomes. Results suggested that religiously active youths report higher levels of social capital resources and that the influence of adolescent religiousness on moral outcomes was mediated through social capital resources. Suggestions for further research and implications for faith-based youth development organizations are considered. PMID- 15355161 TI - Children's perceptions of gender discrimination. AB - Children (N = 76; ages 5-10 years) participated in a study designed to examine perceptions of gender discrimination. Children were read scenarios in which a teacher determined outcomes for 2 students (1 boy and 1 girl). Contextual information (i.e., teacher's past behavior), the gender of the target of discrimination (i.e., student), and the gender of the perpetrator (i.e., teacher) were manipulated. Results indicated that older children were more likely than younger children to make attributions to discrimination when contextual information suggested that it was likely. Girls (but not boys) were more likely to view girls than boys as victims of discrimination, and children with egalitarian gender attitudes were more likely to perceive discrimination than were their peers. PMID- 15355162 TI - Family income and its relation to preschool children's adjustment for families in the NICHD Study of Early Child Care. AB - The current study examines relations of mean-level estimates, linear changes, and instability in income and family processes to child outcomes and addresses whether income, through its impact on family functioning, matters more for children living in poverty. Temporal changes and instability in family processes, but not income, predicted children's adjustment. Cross-sectional mediational analyses indicated that for families living at the poverty threshold, family processes fully mediated the effect of average income over the study period on social behavior but only partially mediated its effect on cognitive-linguistic development. The strength of these associations diminished as average income exceeded the poverty threshold. That is, income had a greater impact on the family functioning and development of poor children than of nonpoor children. PMID- 15355163 TI - Multiple labels for objects in conversations with young children: parents' language and children's developing expectations about word meanings. AB - Children sometimes seem to expect words to have mutually exclusive meanings in certain contexts of early word learning. In 2 studies, 12- to 24-month-old children and their parents were videotaped as they engaged in conversations while playing with sets of toys (sea creatures, vehicles, doll clothing) in free-play, storytelling, and categorization contexts. In both studies, parents demonstrated a reliable preference to provide just 1 label for a given object. Importantly, parents' violations of this preference were usually accompanied by clarifying (or "bridging") information that either indicated the relation between the 2 labels or suggested that 1 of the labels was appropriate. Further, in some contexts, parents' tendency to use multiple labels and to provide bridging information for multiple labels was correlated with children's productive vocabulary. It is argued that these findings support a socio-pragmatic hypothesis about the origins of children's early beliefs about word meanings. PMID- 15355164 TI - Reality compared with its alternatives: age differences in judgments of regret and relief. AB - Three experiments examined developmental change in children's understanding of regret and relief, two second-order emotions whose quality depends on a comparison between reality and "what might have been." In Experiment 1, participants 7 years of age and older, but not 5-year-olds, made regret-related emotion-response judgments that took into account a comparison of reality with its alternatives. In Experiment 2, 5-year-olds judged that an individual would feel better, rather than worse, when a counterfactual outcome was better than what actually occurred (the opposite of the pattern found with older children and adults). Experiment 3 focused on the understanding of relief. In contrast to the findings from Experiment I, the 7-year-olds in Experiment 3 made their judgments solely on the basis of what actually occurred. PMID- 15355165 TI - The emotional integration of childhood experience: physiological, facial expressive, and self-reported emotional response during the adult attachment interview. AB - Attachment researchers claim that individual differences in how adults talk about their early memories reflect qualitatively distinct organizations of emotion regarding childhood experiences with caregivers. Testing this assumption, the present study examined the relationship between attachment dimensions and physiological, facial expressive, as well as self-reported emotional responses during the Adult Attachment Interview (AAI). Consistent with theoretical predictions, more prototypically secure adults behaviorally expressed and reported experiencing emotion consistent with the valence of the childhood events they described. Insecure adults also showed distinctive and theoretically anticipated forms of emotional response: Dismissing participants evidenced increased electrodermal activity during the interview, a sign of emotional suppression, whereas preoccupied adults showed reliable discrepancies between the valence of their inferred childhood experiences and their facial expressive as well as reported emotion during the AAI. Results substantiate a case that the AAI reflects individual differences in emotion regulation that conceptually parallel observations of attachment relationships in infancy. PMID- 15355166 TI - The longitudinal relations of regulation and emotionality to quality of Indonesian children's socioemotional functioning. AB - Data regarding individual differences in children's regulation, emotionality, quality of socioemotional functioning, and shyness were obtained from teachers and peers for 112 Indonesian 6th graders. Similar data (plus parents' reports) also were collected when these children were in 3rd grade. For boys, regulation and low negative emotionality generally predicted positive socioemotional functioning (e.g., social skills, adjustment, prosocial tendencies and peer liking, sympathy) within and across time and across reporters, even at the follow up when initial levels of regulation or negative emotionality were controlled. For girls, relations were obtained primarily for concurrent teacher reports, probably because girls tended to be fairly well regulated and socially competent and variability in their scores was relatively low. Shyness for both sexes tended to be associated with concurrent measures of low regulation, high negative emotionality, and low quality of social competence. PMID- 15355167 TI - Genetic and environmental contributions to general cognitive ability through the first 16 years of life. AB - The genetic and environmental contributions to the development of general cognitive ability throughout the first 16 years of life were examined using sibling data from the Colorado Adoption Project. Correlations were analyzed along with structural equation models to characterize the genetic and environmental influences on longitudinal stability and instability. Intraclass correlations reflected both considerable genetic influence at each age and modest shared environmental influence within and across ages. Modeling results suggested that genetic factors mediated phenotypic stability throughout this entire period, whereas most age-to-age instability appeared to be due to nonshared environmental influences. PMID- 15355168 TI - Estimating retest effects in longitudinal assessments of cognitive functioning in adults between 18 and 60 years of age. AB - Several analyses were conducted on data from samples of adults between 18 and 58 years of age who completed the same cognitive tests after an interval ranging from less than 1 week to 35 years. Because the retest interval varied across individuals, it was possible to determine the length of time needed before the gains associated with a retest decreased to 0 and to obtain simultaneous estimates of the magnitude of effects associated with increased age and a prior assessment. The results indicated that for adults within this age range, 7 or more years were needed before positive retest effects were no longer detectable. Age effects in longitudinal comparisons could be interpreted in terms of large positive effects associated with a prior assessment and negative effects associated with age that were comparable in magnitude to those observed in cross sectional comparisons. PMID- 15355169 TI - Body image among adolescent girls and boys: a longitudinal study. AB - This longitudinal study of adolescent girls and boys examined the contributions of social (peer appearance context), psychological (internalized appearance ideals and appearance social comparison), and biological (body mass) factors to the development of body dissatisfaction. Students (165 girls and 139 boys) completed questionnaires when they were either in 7th grade or 10th grade and again 1 year later. The results for the boys revealed a singular pathway to body dissatisfaction through internalized commitment to muscularity ideals. The prospective analyses of change in body dissatisfaction among the girls reflected the contributions of appearance conversations with friends, appearance social comparisons, and body mass. There was no evidence of mediation among the boys and limited support for it among the girls. PMID- 15355170 TI - Development of visuospatial short-term memory in the second half of the 1st year. AB - Eighty 5.5- to 12.5-month-old infants participated in 4 delayed-response procedures challenging shortterm visuospatial memory (STVM), 2 that varied the time between presentation and search and 2 that varied the number of locations. Within each type of challenge, 1 task required a gaze response and 1 required a reach response. There was little improvement in STVM performance from 5.5 to 8 months and linear improvement in the percentage correct from 8 to 12 months, with overall STVM performance accounting for 66% of the variance in age. Improvement in searching multiple locations lagged behind improvement in spanning longer delays. Memory scores did not vary for the visual and manual tasks. Perseveration was greatest for reach responses, increased with challenge, and decreased with age. ((c) 2004 APA, all rights reserved) PMID- 15355171 TI - Genetic and environmental mechanisms underlying stability and change in problem behaviors at ages 3, 7, 10, and 12. AB - Maternal ratings on internalizing (INT) and externalizing (EXT) behaviors were collected in a large, population-based longitudinal sample. The numbers of participating twin pairs at ages 3, 7, 10, and 12 were 5,602, 5,115, 2,956, and 1,481, respectively. Stability in both behaviors was accounted for by genetic and shared environmental influences. The genetic contribution to stability (INT: 43%; EXT: 60%) resulted from the fact that a subset of genes expressed at an earlier age was still active at the next time point. A common set of shared environmental factors operated at all ages (INT: 47%; EXT: 34%). The modest contribution of nonshared environmental factors (INT: 10%; EXT: 6%) could not be captured by a simple model. Significant age-specific influences were found for all components, indicating that genetic and environmental factors also contributed to changes in problem behavior. PMID- 15355172 TI - Links between sex-typed time use in middle childhood and gender development in early adolescence. AB - The authors studied sex-typing in the kinds (e.g., sports, handicrafts) and social contexts (same- vs. other-sex companions) of children's free time activities, and the links between sex-typed activities and gender development over 2 years. Participants were 200 White, working- and middle-class children (103 girls, 97 boys; mean age = 10.86 years). In annual home interviews, children rated their self-esteem, gender role attitudes and sex-typed personality qualities, academic interests, and school grades. During 7 nightly phone interviews each year, children reported on their activities. Boys were more sex typed than girls in their peer activities, and children were least sex-typed in their activities with siblings. Sex-typed activities in middle childhood predicted individual differences in gender development in early adolescence. PMID- 15355173 TI - The ghost condition: imitation versus emulation in young children's observational learning. AB - Although observational learning by children may occur through imitating a modeler's actions, it can also occur through learning about an object's dynamic affordances--a process that M. Tomasello (1996) calls "emulation." The relative contributions of imitation and emulation within observational learning were examined in a study with 14- to 26-month-old children. The effectiveness of a "ghost" condition, in which the effective operation of the means apparatus was seen to occur without human agency, was compared with that of a standard modeling procedure in which the child saw an experimenter demonstrate the means action. The ghost condition was as likely to encourage observational learning as was the modeling condition; indeed, performance in the ghost condition was significantly better. The role of emulation in the development of observational learning is discussed in the context of a possible form of goal directedness without agency. PMID- 15355224 TI - Prion genetics: new rules for a new kind of gene. AB - Just as nucleic acids can carry out enzymatic reactions, proteins can be genes. These heritable infectious proteins (prions) follow unique genetic rules that enable their identification: reversible curing, inducible "spontaneous generation," and phenotype surprises. Most prions are based on self-propagating amyloids, depend heavily on chaperones, show strain phenomena and, like other infectious elements, show species barriers to transmission. A recently identified prion is based on obligatory self-activation of an enzyme in trans. Although prions can be detrimental, they may also be beneficial to their hosts. PMID- 15355232 TI - Biology and management of insect pests in North American intensively managed hardwood forest systems. AB - Increasing demand for wood and wood products is putting stress on traditional forest production areas, leading to long-term economic and environmental concerns. Intensively managed hardwood forest systems (IMHFS), grown using conventional agricultural as well as forestry methods, can help alleviate potential problems in natural forest production areas. Although IMHFS can produce more biomass per hectare per year than natural forests, the ecologically simplified, monocultural systems may greatly increase the crop's susceptibility to pests. Species in the genera Populus and Salix comprise the greatest acreage in IMHFS in North America, but other species, including Liquidambar styraciflua and Platanus occidentalis, are also important. We discuss life histories, realized and potential damage, and management options for the most economically influential pests that affect these hardwood species. The substantial inherent challenges associated with pest management in the monocultural environments created by IMHFS are reviewed. Finally, we discuss ways to design IMHFS that may reduce their susceptibility to pests, increase their growth and productivity potential, and create a more sustainable environment. PMID- 15355233 TI - Mosquito behavior and vector control. AB - Effective indoor residual spraying against malaria vectors depends on whether mosquitoes rest indoors (i.e., endophilic behavior). This varies among species and is affected by insecticidal irritancy. Exophilic behavior has evolved in certain populations exposed to prolonged spraying programs. Optimum effectiveness of insecticide-treated nets presumably depends on vectors biting at hours when most people are in bed. Time of biting varies among different malaria vector species, but so far there is inconclusive evidence for these evolving so as to avoid bednets. Use of an untreated net diverts extra biting to someone in the same room who is without a net. Understanding choice of oviposition sites and dispersal behavior is important for the design of successful larval control programs including those using predatory mosquito larvae. Prospects for genetic control by sterile males or genes rendering mosquitoes harmless to humans will depend on competitive mating behavior. These methods are hampered by the immigration of monogamous, already-mated females. PMID- 15355234 TI - The genetics and genomics of the silkworm, Bombyx mori. AB - We review progress in applying molecular genetic and genomic technologies to studies in the domesticated silkworm, Bombyx mori, highlighting its use as a model for Lepidoptera, and in sericulture and biotechnology. Dense molecular linkage maps are being integrated with classical linkage maps for positional cloning and marker-assisted selection. Classical mutations have been identified by a candidate gene approach. Cytogenetic and sequence analyses show that the W chromosome is composed largely of nested full-length long terminal repeat retrotransposons. Z-chromosome-linked sequences show a lack of dosage compensation. The downstream sex differentiation mechanism has been studied via the silkworm homolog of doublesex. Expressed sequence tagged databases have been used to discover Lepidoptera-specific genes, provide evidence for horizontal gene transfer, and construct microarrays. Physical maps using large-fragment bacterial artificial chromosome libraries have been constructed, and whole-genome shotgun sequencing is underway. Germline transformation and transient expression systems are well established and available for functional studies, high-level protein expression, and gene silencing via RNA interference. PMID- 15355235 TI - Tsetse genetics: contributions to biology, systematics, and control of tsetse flies. AB - Tsetse flies (Diptera: Glossinidae) constitute a small, ancient taxon of exclusively hematophagous insects that reproduce slowly and viviparously. Because tsetse flies are the only vectors of pathogenic African trypanosomes, they are a potent and constant threat to humans and livestock over much of sub-Saharan Africa. Despite their low fecundity, tsetse flies demonstrate great resilience, which makes population suppression expensive, transient, and beyond the capacities of private and public sectors to accomplish, except over small areas. Nevertheless, control measures that include genetic methods are under consideration at national and supranational levels. There is a pressing need for sufficient laboratory cultures of tsetse flies and financial support to carry out genetic research. Here we review tsetse genetics from organismal and population points of view and identify some research needs. PMID- 15355236 TI - Folsomia candida (Collembola): a "standard" soil arthropod. AB - Folsomia candida Willem 1902, a member of the order Collembola (colloquially called springtails), is a common and widespread arthropod that occurs in soils throughout the world. The species is parthenogenetic and is easy to maintain in the laboratory on a diet of granulated dry yeast. F. candida has been used as a "standard" test organism for more than 40 years for estimating the effects of pesticides and environmental pollutants on nontarget soil arthropods. However, it has also been employed as a model for the investigation of numerous other phenomena such as cold tolerance, quality as a prey item, and effects of microarthropod grazing on pathogenic fungi and mycorrhizae of plant roots. In this comprehensive review, aspects of the life history, ecology, and ecotoxicology of F. candida are covered. We focus on the recent literature, especially studies that have examined the effects of soil pollutants on reproduction in F. candida using the protocol published by the International Standards Organization in 1999. PMID- 15355237 TI - The mevalonate pathway and the synthesis of juvenile hormone in insects. AB - The mevalonate pathway in insects has two important peculiarities, the absence of the sterol branch and the synthesis of juvenile hormone (JH), that may have influenced the mechanisms of regulation. The data available on these mechanisms indicate that cholesterol does not play a regulatory role and that JH modulates transcript levels of a number of genes of the mevalonate pathway or can influence the translatability and/or stability of the transcripts themselves. These data suggest that the mevalonate pathway in insects can best be interpreted in terms of coordinated regulation, in which regulators act in parallel to a number of enzymes, as occurs in the cholesterol-driven pathway in vertebrates. PMID- 15355238 TI - Chemical ecology of locusts and related acridids. AB - The results and insights from recent research on the chemical ecology of polymorphic acridids are reviewed. Many of the new findings come from studies on the desert locust, Schistocerca gregaria, which has continued to be the primary research insect in most laboratories. Earlier confusion between stimuli associated with phase change and social cohesion has been resolved. The roles of chemotactile and olfactory cues together with tactile and visual stimuli in key locust processes, comprising gregarization, social cohesion, synchronous maturation, mating, oviposition, and maternal transfer of gregarious character, are better understood. Some of the key pheromones of the gregarious phase have been characterized. Chemical communication is also shown to be important in the life style of the solitarious phase. The behavioral pattern and responses of this phase reflect a strong propensity of the species to exploit opportunities under appropriate conditions to form or join the crowd and to gregarize. Outstanding questions are highlighted. PMID- 15355239 TI - The evolution of cotton pest management practices in China. AB - The development of cotton pest management practices in China has followed a pattern seen for many crops that rely heavily on insecticides. Helicoverpa armigera resistance to chemical pesticides resulted in the unprecedented pest densities of the early 1990s. Transgenic cotton that expresses a gene derived from the bacterium Bacillus thuringiensis (Bt) has been deployed for combating H. armigera since 1997. The pest management tactics associated with Bt cotton have resulted in a drastic reduction in insecticide use, which usually results in a significant increase in populations of beneficial insects and thus contributes to the improvement of the natural control of some pests. Risk assessment analyses show that the natural refuges derived from the mixed-planting system of cotton, corn, soybean, and peanut on small-scale, single-family-owned farms play an important function in delaying evolution of cotton bollworm resistance, and that no trend toward Bt cotton resistance has been apparent despite intensive planting of Bt cotton over the past several years. PMID- 15355240 TI - Thysanoptera: diversity and interactions. AB - Published literature on thrips has been dominated by descriptive taxonomy, pest control work, and generalized synecology. The lack of studies examining the detailed biology or autecology of any species limits our understanding of how thrips live and the processes underlying their diversification. Similarly, the phylogenetic inadequacy of thrips classification limits our ability to examine the evolution of biological traits. The extent to which our knowledge of the biology of thrips has increased in recent years is reviewed, such as the behavior of particular species and their interactions with other organisms, including host plant associations, pollination, predation, and natural enemies--factors involved in driving diversification within this order of opportunistic insects. PMID- 15355241 TI - Effects of plants genetically modified for insect resistance on nontarget organisms. AB - Insect resistance, based on Bacillus thuringiensis (Bt) endotoxins, is the second most widely used trait (after herbicide resistance) in commercial genetically modified (GM) crops. Other modifications for insect resistance, such as proteinase inhibitors and lectins, are also being used in many experimental crops. The extensive testing on nontarget plant-feeding insects and beneficial species that has accompanied the long-term and wide-scale use of Bt plants has not detected significant adverse effects. GM plants expressing other insect resistant proteins that have a broader spectrum of activity have been tested on only a limited number of nontarget species. Little is known about the persistence of transgene-derived proteins in soil, with the exception of Bt endotoxins, which can persist in soil for several months. Bt plants appear to have little impact on soil biota such as earthworms, collembolans, and general soil microflora. Further research is required on the effects of GM plants on soil processes such as decomposition. Assessment of nontarget impacts is an essential part of the risk assessment process for insect-resistant GM plants. PMID- 15355242 TI - Invasive phytophagous pests arising through a recent tropical evolutionary radiation: the Bactrocera dorsalis complex of fruit flies. AB - The Bactrocera dorsalis complex of tropical fruit flies (Diptera: Tephritidae: Dacinae) contains 75 described species, largely endemic to Southeast Asia. Within the complex are a small number of polyphagous pests of international significance, including B. dorsalis sensu stricto, B. papayae, B. carambolae, and B. philippinensis. Most species within the complex were described in 1994 and since then substantial research has been undertaken in developing morphological and molecular diagnostic techniques for their recognition. Such techniques can now resolve most taxa adequately. Genetic evidence suggests that the complex has evolved in only the last few million years, and development of a phylogeny of the group is considered a high priority to provide a framework for future evolutionary and ecological studies. As model systems, mating studies on B. dorsalis s.s. and B. cacuminata have substantially advanced our understanding of insect use of plant-derived chemicals for mating, but such studies have not been applied to help resolve the limits of biological species within the complex. Although they are commonly regarded as major pests, there is little published evidence documenting economic losses caused by flies of the B. dorsalis complex. Quantification of economic losses caused by B. dorsalis complex species is urgently needed to prioritize research for quarantine and management. Although they have been documented as invaders, relatively little work has been done on the invasion biology of the complex and this is an area warranting further work. PMID- 15355243 TI - Pheromone-mediated aggregation in nonsocial arthropods: an evolutionary ecological perspective. AB - Although the use of aggregation pheromones has been reported for hundreds of nonsocial arthropod species, the evolutionary ecological aspects of this behavior have received little attention. Despite the elaborate literature on mechanisms, robust data on costs and benefits of aggregation pheromones are scant. Existing literature indicates that, in contrast to the diversity of mechanisms, the ecological conditions in which aggregation pheromones are used are more alike. This points to a few general categories for costs and benefits of aggregation pheromones, and these are discussed. We subsequently review interspecific interactions that may be affected by the use of aggregation pheromones. We encounter a strikingly frequent association of aggregation pheromones with fungi and microorganisms and address cross-attraction by competitor species and exploitation by natural enemies. We show that aggregative behavior by individuals through the use of pheromones can profoundly affect ecological interactions and advocate further evolutionary and ecological investigations of pheromone-mediated aggregation. PMID- 15355244 TI - Egg dumping in insects. AB - Females that place eggs under the care of conspecifics have been labeled egg dumpers. Egg dumping is an effective reproductive alternative that lowers risks for, and has the potential to increase fecundity in, its practitioners. Although insect egg dumpers can be social parasites of the maternal behavior of egg recipients, dumping is more likely to be a viable reproductive alternative when the costs to egg recipients are low and thus the defense by potential hosts against egg dumping intrusions is minimal. These conditions are met in insects that guard only eggs or in insects whose eggs hatch into self-supporting precocial young that need little beyond defense from parents. When this is the case, egg dumping is favored by natural and/or kin selection as a mechanism by which dumpers can avoid parental risks and increase fecundity, and egg recipients can enhance offspring survival by diluting predation. PMID- 15355245 TI - Tyramine and octopamine: ruling behavior and metabolism. AB - Octopamine (OA) and tyramine (TA) are the invertebrate counterparts of the vertebrate adrenergic transmitters. They are decarboxylation products of the amino acid tyrosine, with TA as the biological precursor of OA. Nevertheless, both compounds are independent neurotransmitters that act through G protein coupled receptors. OA modulates a plethora of behaviors and peripheral and sense organs, enabling the insect to respond correctly to external stimuli. Because these two phenolamines are the only biogenic amines whose physiological significance is presumably restricted to invertebrates, pharmacologists have focused their attention on the corresponding receptors, which are still believed to represent promising targets for new insecticides. Recent progress made on all levels of OA and TA research has enabled researchers to understand better the molecular events underlying the control of complex behaviors. PMID- 15355246 TI - Evolutionary and mechanistic theories of aging. AB - Senescence (aging) is defined as a decline in performance and fitness with advancing age. Senescence is a nearly universal feature of multicellular organisms, and understanding why it occurs is a long-standing problem in biology. Here we present a concise review of both evolutionary and mechanistic theories of aging. We describe the development of the general evolutionary theory, along with the mutation accumulation, antagonistic pleiotropy, and disposable soma versions of the evolutionary model. The review of the mechanistic theories focuses on the oxidative stress resistance, cellular signaling, and dietary control mechanisms of life span extension. We close with a discussion of how an approach that makes use of both evolutionary and molecular analyses can address a critical question: Which of the mechanisms that can cause variation in aging actually do cause variation in natural populations? PMID- 15355247 TI - Ecological, behavioral, and biochemical aspects of insect hydrocarbons. AB - This review covers selected literature from 1982 to the present on some of the ecological, behavioral, and biochemical aspects of hydrocarbon use by insects and other arthropods. Major ecological and behavioral topics are species- and gender recognition, nestmate recognition, task-specific cues, dominance and fertility cues, chemical mimicry, and primer pheromones. Major biochemical topics include chain length regulation, mechanism of hydrocarbon formation, timing of hydrocarbon synthesis and transport, and biosynthesis of volatile hydrocarbon pheromones of Lepidoptera and Coleoptera. In addition, a section is devoted to future research needs in this rapidly growing area of science. PMID- 15355306 TI - GCN2 phosphorylation of eIF2alpha activates NF-kappaB in response to UV irradiation. AB - In response to UV irradiation, mammalian cells elicit a gene expression programme designed to repair damage and control cell proliferation and apoptosis. Important members of this stress response include the NF-kappaB (nuclear factor-kappaB) family. However, the mechanisms by which UV irradiation activates NF-kappaB are not well understood. In eukaryotes, a variety of environmental stresses are recognized and remediated by a family of protein kinases that phosphorylate the alpha subunit of eIF2 (eukaryotic initiation factor-2). In the present study we show that NF-kappaB in MEF (murine embryo fibroblast) cells is activated by UV-C and UV-B irradiation through a mechanism requiring eIF2alpha phosphorylation. The primary eIF2alpha kinase in response to UV is GCN2 (general control non derepressible-2), with PEK/PERK (pancreatic eIF2alpha kinase/RNA-dependent protein-kinase-like endoplasmic-reticulum kinase) carrying out a secondary function. Our studies indicate that lowered protein synthesis accompanying eIF2alpha phosphorylation, combined with eIF2alpha kinase-independent turnover of IkappaBalpha (inhibitor of kappaBalpha), reduces the levels of IkappaBalpha in response to UV irradiation. Release of NF-kappaB from the inhibitory IkappaBalpha would facilitate NF-kappaB entry into the nucleus and targeted transcriptional control. We also find that loss of GCN2 in MEF cells significantly enhances apoptosis in response to UV exposure similar to that measured in cells deleted for the RelA/p65 subunit of NF-kappaB. These results demonstrate that GCN2 is central to recognition of UV stress, and that eIF2alpha phosphorylation provides resistance to apoptosis in response to this environmental insult. PMID- 15355307 TI - The septin Sept5/CDCrel-1 competes with alpha-SNAP for binding to the SNARE complex. AB - SNARE (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor) proteins are supposed to mediate the docking and/or fusion of the vesicle with the plasma membrane. However, it is not clearly understood how this process is regulated. In a search for potential SNARE regulators, we recently identified septin 5 (Sept5) as a novel SNARE interacting protein. Septins were first identified as filamentous proteins required for cytokinesis in yeast. Several septins have now been identified in mammals but little is known about their functions. We have previously shown that Sept5 is predominantly expressed in the brain, where it associates with vesicles and membranes through its interaction with the SNARE domain of syntaxin 1A. Furthermore, Sept5 appears to inhibit exocytosis, possibly by regulating vesicle targeting and/or fusion events. To gain insight into the role of Sept5, we have mapped the Sept5 domains important for syntaxin binding. We also investigated the ability of Sept5 to bind to syntaxin when in various protein complexes. Although Sept5 cannot bind an nSec1-syntaxin complex, it can bind syntaxin in a SNARE complex. This interaction is occluded by the binding of alpha-SNAP, suggesting that Sept5 may regulate the availability of SNARE proteins through its interaction with syntaxin and the 7 S complex. PMID- 15355308 TI - Regulation of all members of the antizyme family by antizyme inhibitor. AB - ODC (ornithine decarboxylase) is the rate-limiting enzyme in polyamine biosynthesis. Polyamines are essential for cellular growth and differentiation but enhanced ODC activity is associated with cell transformation. Post translationally, ODC is negatively regulated through members of the antizyme family. Antizymes inhibit ODC activity, promote ODC degradation through the 26 S proteasome and regulate polyamine transport. Besides the ubiquitously expressed antizymes 1 and 2, there is the tissue-specific antizyme 3 and an yet uncharacterized antizyme 4. Antizyme 1 has been shown to be negatively regulated through the AZI (antizyme inhibitor) that binds antizyme 1 with higher affinity compared with ODC. In the present study, we show by yeast two- and three-hybrid protein-protein interaction studies that AZI interacts with all members of the antizyme family and is capable of disrupting the interaction between each antizyme and ODC. In a yeast-based ODC complementation assay, we show that human ODC is able to complement fully the function of the yeast homologue of ODC. Co expression of antizymes resulted in ODC inhibition and cessation of yeast growth. The antizyme-induced growth inhibition could be reversed by addition of putrescine or by the co-expression of AZI. The protein interactions could be confirmed by immunoprecipitation of the human ODC-antizyme 2-AZI complexes. In summary, we conclude that human AZI is capable of acting as a general inhibitor for all members of the antizyme family and that the previously not yet characterized antizyme 4 is capable of binding ODC and inhibiting its enzymic activity similar to the other members of the antizyme family. PMID- 15355309 TI - The predominant polyphenol in the leaves of the resurrection plant Myrothamnus flabellifolius, 3,4,5 tri-O-galloylquinic acid, protects membranes against desiccation and free radical-induced oxidation. AB - The predominant (>90%) low-molecular-mass polyphenol was isolated from the leaves of the resurrection plant Myrothamnus flabellifolius and identified to be 3,4,5 tri-O-galloylquinic acid using 1H and 13C one- and two-dimensional NMR spectroscopy. The structure was confirmed by mass spectrometric analysis. This compound was present at high concentrations, 44% (by weight) in hydrated leaves and 74% (by weight) in dehydrated leaves. Electron microscopy of leaf material fixed with glutaraldehyde and caffeine demonstrated that the polyphenols were localized in large vacuoles in both hydrated and dehydrated leaves. 3,4,5 Tri-O galloylquinic acid was shown to stabilize an artificial membrane system, liposomes, against desiccation if the polyphenol concentration was between 1 and 2 microg/mug phospholipid. The phase transition of these liposomes observed at 46 degrees C was markedly diminished by the presence of 3,4,5 tri-O-galloylquinic acid, suggesting that the presence of the polyphenol maintained the membranes in the liquid crystalline phase at physiological temperatures. 3,4,5 Tri-O galloylquinic acid was also shown to protect linoleic acid against free radical induced oxidation. PMID- 15355310 TI - Axon behaviour at Schwann cell - astrocyte boundaries: manipulation of axon signalling pathways and the neural adhesion molecule L1 can enable axons to cross. AB - Axon regeneration in vivo is blocked at boundaries between Schwann cells and astrocytes, such as occur at the dorsal root entry zone and around peripheral nerve or Schwann cell grafts. We have created a tissue culture model of these boundaries in Schwann cell - astrocyte monolayer co-cultures. Axon behaviour resembles that in vivo, with axons showing a strong preference for Schwann cells over astrocytes. At boundaries between the two cell types, axons growing on astrocytes cross readily onto Schwann cells, but only 15% of axons growing on Schwann cells are able to cross onto astrocytes. Treatment with chondroitinase or chlorate to reduce inhibition by proteoglycans did not change this behaviour. The neural adhesion molecule L1 is present on Schwann cells and not astrocytes, and manipulation of L1 by application of an antibody, L1-Fc in solution, or adenoviral transduction of L1 into astrocytes increased the proportion of axons able to cross onto astrocytes to 40-50%. Elevating cAMP levels increased crossing from Schwann cells onto astrocytes in live and fixed cultures, and had a co operative effect with NT-3 but not with NGF. Inactivation of Rho with a cell permeant form of C3 exoenzyme also increased crossing from Schwann cells to astrocytes. Our experiments indicate that the preference of axons for Schwann cells is largely mediated by the presence of L1 on Schwann cells but not astrocytes, and that manipulation of growth cone signalling pathways can allow axons to disregard boundaries between the two cell types. PMID- 15355311 TI - Binding partners L1 cell adhesion molecule and the ezrin-radixin-moesin (ERM) proteins are involved in development and the regenerative response to injury of hippocampal and cortical neurons. AB - Regeneration of the adult central nervous system may require recapitulation of developmental events and therefore involve the re-expression of developmentally significant proteins. We have investigated whether the L1 cell adhesion molecule, and its binding partner, the ezrin-radixin-moesin (ERM) proteins are involved in the neuronal regenerative response to injury. Hippocampal and cortical neurons were cultured in vitro on either an L1 substrate or poly-L-lysine, and ERM and other neuronal proteins were localized immunocytochemically both developmentally and following neurite transection of neurons maintained in long-term culture. Activated ERM was localized to growth cones up to 7 days in vitro but relatively mature cultures (21 days in vitro) were devoid of active ERM proteins. However, ERM proteins were localized to the growth cones of sprouting neuronal processes that formed several hours after neurite transection. In addition, the L1 substrate, relative to poly-L-lysine, resulted in significantly longer regenerative neurites, as well as larger growth cones with more filopodia. Furthermore, neurons derived from the cortex formed significantly longer post injury neurite sprouts at 6 h post-injury than hippocampal derived neurons grown on both substrates. We have demonstrated that L1 and the ERM proteins are involved in the neuronal response to injury, and that neurons derived from the hippocampus and cortex may have different post-injury regenerative neurite sprouting abilities. PMID- 15355312 TI - The remyelinating potential and in vitro differentiation of MOG-expressing oligodendrocyte precursors isolated from the adult rat CNS. AB - There is a long-standing controversy as to whether oligodendrocytes may be capable of cell division and thus contribute to remyelination. We recently published evidence that a subpopulation of myelin oligodendrocyte glycoprotein (MOG)-expressing cells in the adult rat spinal cord co-expressed molecules previously considered to be restricted to oligodendrocyte progenitors [G. Li et al. (2002) Brain Pathol., 12, 463-471]. To further investigate the properties of MOG-expressing cells, anti-MOG-immunosorted cells were grown in culture and transplanted into acute demyelinating lesions. The immunosorting protocol yielded a cell preparation in which over 98% of the viable cells showed anti-MOG- and O1 immunoreactivity; 12-15% of the anti-MOG-immunosorted cells co-expressed platelet derived growth factor alpha receptor (PDGFRalpha) or the A2B5-epitope. When cultured in serum-free medium containing EGF and FGF-2, 15-18% of the anti-MOG immunosorted cells lost anti-MOG- and O1-immunoreactivity and underwent cell division. On removal of these growth factors, cells differentiated into oligodendrocytes, or astrocytes and Schwann cells when the differentiation medium contained BMPs. Transplantation of anti-MOG-immunosorted cells into areas of acute demyelination immediately after isolation resulted in the generation of remyelinating oligodendrocytes and Schwann cells. Our studies indicate that the adult rat CNS contains a significant number of oligodendrocyte precursors that express MOG and galactocerebroside, molecules previously considered restricted to mature oligodendrocytes. This may explain why myelin-bearing oligodendrocytes were considered capable of generating remyelinating cells. Our study also provides evidence that the adult oligodendrocyte progenitor can be considered as a source of the Schwann cells that remyelinate demyelinated CNS axons following concurrent destruction of oligodendrocytes and astrocytes. PMID- 15355314 TI - Developmental expression of the Ca2+-binding proteins calretinin and parvalbumin at the calyx of Held of rats and mice. AB - Ca(2+)-binding proteins of the EF-hand family are widely expressed in the CNS, and contribute to intracellular Ca(2+) buffering in neurons. In nerve terminals, Ca(2+)-binding proteins are likely to regulate transmitter release probability and synaptic short-term-plasticity. Here, we investigated the developmental expression pattern of calretinin and parvalbumin at a large excitatory synapse, the calyx of Held in the medial nucleus of the trapezoid body (MNTB) of rats and mice. We used two-colour immunofluorescence imaging with primary antibodies detecting one of the Ca(2+)-binding proteins, and a presynaptic marker protein, Rab-3A. Calretinin was found in nerve terminals of the calyx of Held, but not in postsynaptic principal cells. The presynaptic density of Calretinin staining, and the degree of colocalization with Rab-3A increased during postnatal development (P6-P31). Surprisingly, not all calyces of Held expressed calretinin. In rats, calretinin-containing calyces were irregularly interspersed with calretinin negative calyces, whereas in mice, calretinin-positive calyces were preferentially located in the lateral portion of the MNTB. The percentage of calretinin-positive calyces increased during development, to about 75% and 20% at P30 in rats and in mice, respectively. Parvalbumin was present in the presynaptic calyces of Held and in the nerve fibres entering the MNTB, as well as in the somata of the MNTB principal neurons. An up-regulation of calretinin and parvalbumin in calyces of Held probably increases the presynaptic Ca(2+) buffering strength during postnatal development, but the unexpected heterogeneity of calretinin expression might cause differences in Ca(2+) signalling and transmitter release probability between calyces of Held. PMID- 15355313 TI - The effect of PARP inhibitor on ischaemic cell death, its related inflammation and survival signals. AB - Poly(ADP-ribose) polymerase (PARP) plays an important role in ischaemic cell death, and 3-aminobenzamide (3-AB), one of the PARP inhibitors, has a protective effect on ischaemic stroke. We investigated the neuroprotective mechanisms of 3 AB in ischaemic stroke. The occlusion of middle cerebral artery (MCA) was made in 170 Sprague-Dawley rats, and reperfusion was performed 2 h after the occlusion. Another 10 Sprague-Dawley rats were used for sham operation. 3-AB was administered to 85 rats 10 min before the occlusion [3-AB group (n = 85) vs. control group without 3-AB (n = 85)]. Infarct volume and water content were measured, brain magnetic resonance imaging, terminal deoxynucleotidyltransferase (TdT)-mediated dUTP-biotin nick end-labelling (TUNEL) and Cresyl violet staining were performed, and immunoreactivities (IRs) of poly(ADP-ribose) polymer (PAR), cleaved caspase-3, CD11b, intercellular adhesion molecule-1 (ICAM-1), cyclooxygenase-2 (COX-2), phospho-Akt (pAkt) and phospho-glycogen synthase kinase 3 (pGSK-3) were compared in the peri-infarcted region of the 3-AB group and its corresponding ischaemic region of the control group at 2, 8, 24 and 72 h after the occlusion. In the 3-AB group, the infarct volume and the water content were decreased (about 45% and 3.6%, respectively, at 24 h), the number of TUNEL positive cells was decreased (about 36% at 24 h), and the IRs of PAR, cleaved caspase-3, CD11b, ICAM-1 and COX-2 were significantly reduced, while the IRs of pAkt and pGSK-3 were increased. These results suggest that 3-AB treatment could reduce the infarct volume by reducing ischaemic cell death, its related inflammation and increasing survival signals. The inhibition of PARP could be another potential neuroprotective strategy in ischaemic stroke. PMID- 15355315 TI - Mutation of the feh-1 gene, the Caenorhabditis elegans orthologue of mammalian Fe65, decreases the expression of two acetylcholinesterase genes. AB - The molecular adaptor Fe65 is one of the cytosolic ligands of the Alzheimer's beta-amyloid precursor protein (APP), and this complex is believed to play important roles in mammalian cells. Upon cleavage of APP by specific processing activities, the complex between Fe65 and the APP intracellular domain (AICD) translocates to the nucleus. Experimental evidence suggests that the Fe65-AICD complex regulates gene transcription. In Caenorhabditis elegans the orthologue of the Fe65 gene, feh-1, regulates pharyngeal activity. In fact, the rate of pharyngeal contraction is increased following transient or stable suppression of the feh-1 gene expression. Here we show that the increased contraction rate of the pharynx in feh-1 mutant worms is associated to decreased acetylcholinesterase activity. The decreased activity is accompanied by reduced expression of ace-1 and ace-2 transcripts, coding for the two major acetylcholinesterase activities in the nematode. These results indicate a target of the regulatory mechanisms based on the Fe65-APP complex that could be relevant for the pathogenesis of Alzheimer's disease. PMID- 15355316 TI - Angiotensin II increases differentiation of dopaminergic neurons from mesencephalic precursors via angiotensin type 2 receptors. AB - In addition to the well-known actions of the humoral renin-angiotensin system, all components of this system are present in many tissues, including the brain, and may play a major role in brain development and differentiation. We investigated the possible effects of angiotensin II on the generation of dopaminergic phenotype neurons from proliferating neurospheres of mesencephalic precursors. We observed immunoreactivity for both angiotensin type 1 and type 2 (AT(1) and AT(2)) receptors in the cell aggregates. Double immunolabeling studies revealed that both receptor types are located in neurons and astrocytes. Interestingly, neurons with a dopaminergic phenotype (i.e. tyrosine hydroxylase activity) showed double labeling for AT(1) and AT(2) receptors although the labeling for AT(2) was more intense. Treatment of the neurospheres with angiotensin II (100 nm) during the differentiation period induced a marked increase (about 400%) in the generation of dopaminergic neurons. This was not affected by treatment with the AT(1) antagonist ZD 7155 but was blocked by treatment with the AT(2) antagonist PD 123319. This suggests that AT(2) receptors mediate the stimulatory effect of angiotensin II on the generation of dopaminergic neurons. Apoptotic cell death studies and bromodeoxyuridine immunohistochemistry indicated that the increase in generation of dopaminergic neurons is not due to increased survival or proliferation of dopaminergic cells during treatment with angiotensin and suggested that angiotensin induces increased differentiation of mesencephalic precursors towards the dopaminergic phenotype. Manipulation of the renin-angiotensin system may be useful for increasing production of dopaminergic neurons for transplantation in Parkinson's disease. PMID- 15355317 TI - Unique accumulation of neuropeptides in an insect: FMRFamide-related peptides in the cockroach, Periplaneta americana. AB - FMRFamides belong to the most extensively studied neuropeptides in invertebrates and exhibit diverse physiological effects on different target organs, such as muscles, intestine and the nervous system. This study on the American cockroach confirms for the first time that extended FMRFamides occur in non-dipteran insects. By means of tandem mass spectrometry, these neuropeptides were structurally elucidated, and sequence information was used for subsequent cloning of the cockroach FMRFamide gene. This precursor gene encodes for 24 putative peptides and shows sufficient similarity with the Drosophila FMRFamide gene. Of the 24 peptides, 23 were detected by mass spectrometric methods; it is the highest number of neuropeptide forms shown to be expressed from a single precursor in any insect. The expression was traced back to single neurons in the thoracic ganglia. The unique accumulation of these FMRFamide-related peptides in thoracic perisympathetic organs provides the definite evidence for a tagma specific distribution of peptidergic neurohormones in neurohaemal release sites of the insect CNS. Excitatory effects of the cockroach FMRFamides were observed on antenna-heart preparations. In addition, the newly described FMRFamides reduce the spike frequency of dorsal-unpaired median neurons and reduce the intracellular calcium concentration, which may affect the peripheral release of the biogenic amine octopamine. PMID- 15355318 TI - Cysteinyl-leukotrienes are released from astrocytes and increase astrocyte proliferation and glial fibrillary acidic protein via cys-LT1 receptors and mitogen-activated protein kinase pathway. AB - Cysteinyl-leukotrienes (cys-LTs), potent mediators in inflammatory diseases, are produced by nervous tissue, but their cellular source and role in the brain are not very well known. In this report we have demonstrated that rat cultured astrocytes express the enzymes (5'-lipoxygenase and LTC(4) synthase) required for cys-LT production, and release cys-LTs in resting condition and, to a greater extent, in response to calcium ionophore A23187, 1 h combined oxygen-glucose deprivation or 2-methyl-thioATP, a selective P2Y(1)/ATP receptor agonist. MK-886, a LT synthesis inhibitor, prevented basal and evoked cys-LT release. In addition, 2-methyl-thioATP-induced cys-LT release was abolished by suramin, a P2 receptor antagonist, or by inhibitors of ATP binding cassette proteins involved in cys-LT release. We also showed that astrocytes express cys-LT(1) and not cys-LT(2) receptors. The stimulation of these receptors by LTD(4) activated the mitogen activated protein kinase (MAPK) pathway. This effect was: (i) insensitive to inhibitors of receptor-coupled Gi protein (pertussis toxin) or tyrosine kinase receptors (genistein); (ii) abolished by MK-571, a cys-LT(1) selective receptor antagonist, or PD98059, a MAPK inhibitor; (iii) reduced by inhibitors of calcium/calmodulin-dependent kinase II (KN-93), Ca(2+)-dependent and -independent (GF102903X) or Ca(2+)-dependent (Go6976) protein kinase C isoforms. LTD(4) also increased astrocyte proliferation and glial fibrillary acidic protein content, which are considered hallmarks of reactive astrogliosis. Both effects were counteracted by cell pretreatment with MK-571 or PD98059. Thus, cys-LTs released from astrocytes might play an autocrine role in the induction of reactive astrogliosis that, in brain injuries, contributes to the formation of a reparative glial scar. PMID- 15355319 TI - Monocarboxylate transport inhibition alters retinal function and cellular amino acid levels. AB - We assessed the effect of the in vivo application of monocarboxylate transport inhibitors on retinal function and amino acid immunocytochemistry. We wanted to determine the impact that altered aerobic metabolite availability has on retinal function and the characteristics of amino acid shunting into metabolic pools. Electroretinograms were collected from anaesthetized rats at various times after intravitreal injection of the monocarboxylate transport inhibitors alpha-cyano-4 hydroxycinnamate (4-CIN; 2 micro L, 0.1-10 mm) or p-(dipropylsulphamoyl)benzoic acid (probenecid; 1-10 mm). Changes in retinal function were compared with quantitative amino acid immunocytochemical changes in retinas harvested 20 and 40 min after either 4-CIN or vehicle treatment. The injection of 4-CIN resulted in a dose-dependent reduction of the ON-bipolar cell P2 wave amplitude (20-80%) and delay in its implicit time. The phototransduction sensitivity was mildly reduced whereas the ON-bipolar cell P2 sensitivity was unaffected. Probenecid induced functional changes similar to those observed with 4-CIN. We also mapped the amino acid alterations within specific cell classes induced by 4-CIN application. All neurones displayed a reduced glutamate content averaging 48%; reduced GABA (31%) and glycine (28%) were found within amacrine cells and glutamine was reduced in all cell classes except photoreceptor and Muller cells. All cell classes in the retina demonstrated increases in aspartate (57%), whereas leucine (24%) and ornithine (21%) were only significantly increased in photoreceptor and bipolar cells. The reduction in glutamate immunolabelling in specific retinal cell classes was mirrored by an increase in aspartate levels at these locations. In addition, attenuated glutamine immunolabelling also closely matched the spatial pattern observed for glutamate. Our immunocytochemical analysis provides evidence that monocarboxylate transport inhibition induces a shift in the equilibrium of glutamate transamination reactions involving aspartate throughout the retina whereas photoreceptor and bipolar cells also use glutamate transamination reactions involving ornithine and leucine. The distribution pattern of glutamine secondary to monocarboxylate inhibition suggests that this amino acid is a major precursor for glutamate throughout the retina. PMID- 15355320 TI - Reduced plasticity of cortical whisker representation in adult tenascin-C deficient mice after vibrissectomy. AB - The effect of the extracellular matrix recognition molecule tenascin-C on cerebral plasticity induced by vibrissectomy was investigated with 2-deoxyglucose (2DG) brain mapping in tenascin-C-deficient mice. Unilateral vibrissectomy sparing row C of vibrissae was performed in young adult mice. Two months later, cortical representations of spared row C vibrissae and control row C on the other side of the snout were visualized by [(14)C]2DG autoradiography. In both wild type and tenascin-C-deficient mice, cortical representation of the spared row was expanded in all layers of the barrel cortex. However, the effect was significantly more extensive in wild-type animals than in the mutant. Elimination of tenascin-C by genetic manipulation thus reduces the effect of vibrissectomy observed in the somatosensory cortex. No increase in number of fibres in the vibrissal nerve of spared vibrissae was seen, and occurrence of additional nerve to the spared follicle was very rare. Thus, in tenascin-C-deficient mice functional plasticity seems to be impaired within the CNS. PMID- 15355321 TI - Rapid assessment of in vivo cholinergic transmission by amperometric detection of changes in extracellular choline levels. AB - Conventional microdialysis methods for measuring acetylcholine (ACh) efflux do not provide sufficient temporal resolution to relate cholinergic transmission to individual stimuli or behavioral responses, or sufficient spatial resolution to investigate heterogeneities in such regulation within a brain region. In an effort to overcome these constraints, we investigated a ceramic-based microelectrode array designed to measure amperometrically rapid changes in extracellular choline as a marker for cholinergic transmission in the frontoparietal cortex of anesthetized rats. These microelectrodes exhibited detection limits of 300 nm for choline and selectivity (> 100 : 1) of choline over interferents such as ascorbic acid. Intracortical pressure ejections of choline (20 mm, 66-400 nL) and ACh (10 and 100 mm, 200 nL) dose-dependently increased choline-related signals that were cleared to background levels within 10 s. ACh, but not choline-induced signals, were significantly attenuated by co ejection of the acetylcholinesterase inhibitor neostigmine (Neo; 100 mm). Pressure ejections of drugs known to increase cortical ACh efflux, potassium (KCl; 70 mm, 66, 200 nL) and scopolamine (Scop; 10 mm, 200 nL), also markedly increased extracellular choline signals, which again were inhibited by Neo. Scop induced choline signals were also found to be tetrodotoxin-sensitive. Collectively, these findings suggest that drug-induced increases in current measured with these microelectrode arrays reflect the oxidation of choline that is neuronally derived from the release and subsequent hydrolysis of ACh. Choline signals assessed using enzyme-selective microelectrode arrays may represent a rapid, sensitive and spatially discrete measure of cholinergic transmission. PMID- 15355322 TI - Adrenoceptor subtype-specific acceleration of the hypoxic depression of excitatory synaptic transmission in area CA1 of the rat hippocampus. AB - The depression of excitatory synaptic transmission by hypoxia in area CA1 of the hippocampus is largely dependent upon the activation of adenosine A(1) receptors on presynaptic glutamatergic terminals. As well as adenosine, norepinephrine levels increase in the hypoxic/ischemic hippocampus. We sought to determine the influence of alpha- and beta-adrenoceptor (AR) activation on the hypoxic depression of synaptic transmission utilizing electrophysiological, pharmacological and adenosine sensor techniques. Norepinephrine depressed synaptic transmission and significantly accelerated the hypoxic depression of synaptic transmission. The alpha-AR agonist 6-fluoronorepinephrine mimicked both of these effects whilst the alpha(2)-AR antagonist yohimbine, but not the alpha(1)-AR antagonist urapidil, prevented the actions of 6-fluoronorepinephrine. In contrast, the beta-AR agonist isoproterenol enhanced synaptic transmission and only accelerated the hypoxic depression of transmission in hypoxia-conditioned slices in which the hypoxic release of adenosine is reduced. The effects of isoproterenol were blocked by the non-selective beta-AR antagonist propranolol and the selective beta(1)-AR antagonist betaxolol. Using an enzyme-based adenosine sensor we observed that the application of the beta-AR agonist resulted in increased extracellular adenosine during repeated hypoxia. Our results suggest that alpha(2)-AR activation facilitates the hypoxic depression of synaptic transmission probably via the known alpha(2)-AR-mediated inhibition of presynaptic calcium channels whereas beta(1)-AR activation does so via increased extracellular adenosine and greater activation of inhibitory adenosine A(1) receptors. PMID- 15355323 TI - Dual effects of gabapentin and pregabalin on glutamate release at rat entorhinal synapses in vitro. AB - We have recently shown that the anticonvulsant drugs phenytoin, lamotrigine and sodium valproate all reduce the release of glutamate at synapses in the entorhinal cortex in vitro. In the present investigation we determined whether this property was shared by gabapentin and pregabalin, using whole-cell patch clamp recordings of excitatory postsynaptic currents (EPSCs) in layer V neurons in slices of rat entorhinal cortex. Both drugs reduced the amplitude and increased the paired-pulse ratio of EPSCs evoked by electrical stimulation of afferent inputs, suggesting a presynaptic effect to reduce glutamate release. The frequency of spontaneous EPSCs (sEPSCs) was concurrently reduced by GBP, further supporting a presynaptic action. There was no significant change in amplitude although a slight reduction was seen, particularly with gabapentin, which may reflect a reduction in the number of larger amplitude sEPSCs. When activity independent miniature EPSCs were recorded in the presence of tetrodotoxin, both drugs continued to reduce the frequency of events with no change in amplitude. The reduction in frequency induced by gabapentin or pregabalin was blocked by application of the l-amino acid transporter substrate l-isoleucine. The results show that gabapentin and pregabalin, like other anticonvulsants, reduce glutamate release at cortical synapses. It is possible that this reduction is a combination of two effects: a reduction of activity-dependent release possibly via interaction with P/Q-type voltage-gated Ca channels, and a second action, as yet unidentified, occurring downstream of Ca influx into the presynaptic terminals. PMID- 15355324 TI - Deletion of the CCK2 receptor gene reduces mechanical sensitivity and abolishes the development of hyperalgesia in mononeuropathic mice. AB - Previous studies suggest that cholecystokinin (CCK) is implicated in the modulation of pain sensitivity and the development of neuropathic pain. We used CCK(2) receptor deficient (CCK(2) (-/-)) mice and assessed their mechanical sensitivity using Von Frey filaments, as well as the development and time course of mechanical hyperalgesia in a model of neuropathic pain. We found that CCK(2) ( /-) mice displayed mechanical hyposensitivity, which was reversed to the level of wild-type animals after administration of naloxone (0.1-10 mg/kg). On the other hand, injection of L-365260 (0.01-1 mg/kg), an antagonist of CCK(2) receptors, decreased dose-dependently, mechanical sensitivity in wild-type mice. The mechanism of reduced mechanical sensitivity in CCK(2) (-/-) mice may be explained by changes in interactions between CCK and opioid systems. Indeed, CCK(2) (-/-) mice natively expressed higher levels of lumbar CCK(1), opioid delta and kappa receptors. Next, we found that CCK(2) (-/-) mice did not develop mechanical hyperalgesia in the Bennett's neuropathic pain model. Induction of neuropathy resulted in decrease of lumbar pro-opiomelanocortin (POMC) gene expression in wild-type mice, but increase of POMC expression in CCK(2) (-/-) mice. In addition, induction of neuropathy resulted in further increase of opioid delta receptor in CCK(2) (-/-) mice. Gene expression results indicate up-regulation of opioid system in CCK(2) (-/-) mice, which apparently result in decreased neuropathy score. Our study suggests that not only pain sensitivity, but also mechanical sensitivity and the development of neuropathic pain are regulated by antagonistic interactions between CCK and opioid systems. PMID- 15355325 TI - Striatal dopamine D2/D3 receptor availability correlates with individual response characteristics to pain. AB - We studied in healthy humans the contribution of cerebral dopamine D2/D3 receptors to individual differences in response characteristics to painful stimulation. Positron emission tomography was used to measure the dopamine D2/D3 binding potential (D2/D3 BP) with [(11)C]raclopride in the striatum (n = 8) and with [(11)C]FLB 457 in the extrastriatal regions (n = 11). Sensitivity to cutaneous heat pain was assessed by a traditional threshold method and by an analysis based on the signal detection theory which allows the separation of an individual subject's discriminative capacity from the response criterion, i.e. the area under the receiver operating characteristic curve provides a measure of the sensory discriminability (sensory factor) and the response criterion gives an estimate of the subject's response bias or attitude (nonsensory factor). The pain threshold and response criterion were inversely correlated with the D2/D3 BP in the right putamen, whereas the discriminative capacity was not significantly correlated with the D2/D3 BP in any brain region. The correlation of the D2/D3 BP in the putamen with the pain threshold and the subject's response criterion may rather be explained by a dopaminergic effect on nonsensory factors determining the subject's attitude towards pain than by a dopaminergic effect on the subject's discriminative capacity. Alternatively, striatal dopamine D2/D3 receptors could control a modulatory pathway producing a parallel shift in the stimulus-response function for sensory signals, mimicking a change in the subject's response criterion. PMID- 15355326 TI - SNAP-25 in hippocampal CA1 region is involved in memory consolidation. AB - As a synaptosomal protein, SNAP-25 plays a role in a number of neuronal functions including axonal growth, dendrite formation, fusion of synaptic vesicles with membrane and the expression of long-term potentiation (LTP) in the hippocampus. Using a learning/memory behavior screening, we identified SNAP-25 as one of the differentially expressed genes in the hippocampus upon behavioral training. The inhibition of SNAP-25 with intracerebroventricular antisense oligonucleotide caused a deficit in long- but not short-term memory for step-down inhibitory avoidance. Intra-CA1 infusion of the SNAP-25 antisense oligonucleotide impaired long-term contextual fear memory and spatial memory and interfered with the LTP of synaptic transmission in the CA1 region. The inhibitory effect on LTP was not mediated by a pre-synaptic mechanism because paired pulse facilitation of synaptic transmission was not affected after administration of the antisense oligonucleotide. Together, the results suggest that SNAP-25 in the CA1 region is involved in memory consolidation. PMID- 15355327 TI - 3beta-20beta-dihydroxy-5alpha-pregnane (UC1011) antagonism of the GABA potentiation and the learning impairment induced in rats by allopregnanolone. AB - Allopregnanolone is a progesterone metabolite and GABA-A receptor modulator with benzodiazepine like effects, including decreased learning and memory. In vitro 3beta-hydroxypregnane steroids antagonize allopregnanolone-induced effects, but no antagonism has been shown in vivo. Our purpose was to evaluate 3beta-20beta dihydroxy-5alpha-pregnane (UC1011) as a blocker of allopregnanolone-induced effects in vivo and in vitro in rats. We tested adult male Wistar rats in the Morris water maze 8 min after daily injections (i.v.) of allopregnanolone 2 mg/kg (n = 21); allopregnanolone : UC1011 2 : 6 (n = 7), 2 : 8 (n = 7), 2 : 20 (n = 14) mg/kg; UC1011 20 mg/kg (n = 14); or vehicle (10% 2-hydroxypropyl-beta cyclodextrin, n = 4). Studies of chloride ion uptake into cortical and hippocampal membrane preparations were performed. The latency to find the hidden platform was still high in the allopregnanolone-injected group on day 6. Day 3-6 rats injected with allopregnanolone and UC1011 (2 : 20 mg/kg) had lower latency (P < 0.05), compared to the allopregnanolone-injected group. The group that only received UC1011 learned the location of the platform as fast as the controls. There was no significant difference in swim speed between groups. The time spent swimming close to the pool wall was in the allopregnanolone : UC1011 group (2 : 20 mg/kg) significantly decreased (P < 0.05, day 3-6), compared to the allopregnanolone-injected group. The increased chloride ion uptake induced by increasing dosage of allopregnanolone in the presence of 10 micro m GABA was significantly decreased with UC1011 (P < 0.01), in both cortical and hippocampal homogenates. In conclusion, UC1011 can via antagonism at the GABA-A receptor reduce the negative allopregnanolone effect on learning in the water maze. PMID- 15355328 TI - Lesions of the Edinger-Westphal nucleus in C57BL/6J mice disrupt ethanol-induced hypothermia and ethanol consumption. AB - The Edinger-Westphal nucleus (EW) is a brain region that has recently been implicated as an important novel neural target for ethanol. Thus, the EW is the only brain region consistently showing elevated c-Fos expression following both voluntary and involuntary ethanol administration. Ethanol-induced c-Fos expression in the EW has been shown to occur in urocortin I-positive neurons. Moreover, previous reports using several genetic models have demonstrated that differences in the EW urocortin I system are correlated with ethanol-mediated behaviours such as ethanol-induced hypothermia and ethanol consumption. The aim of this study was to confirm these relationships using a more direct strategy. Thus, ethanol responses were measured following electrolytic lesions of the EW in male C57BL/6J mice. Both EW-lesioned and sham-operated animals were tested for several ethanol sensitivity measures and ethanol consumption in a two-bottle choice test. The results show that lesions of the EW significantly disrupted ethanol-induced hypothermia, while having no effect on pupillary dilation, locomotor activity or ethanol-induced sedation. In addition, EW-lesioned animals showed significantly lower ethanol preference and total ethanol dose consumed in the two-bottle choice test. EW-lesioned animals also consumed less sucrose than sham-operated animals, but did not have altered preferences for sucrose or quinine in a two-bottle choice test. These data support previously observed genetic correlations between EW urocortin I expression and both ethanol-induced hypothermia and ethanol consumption. Taken together, the findings suggest that the EW may function as a sensor for ethanol, which can influence ethanol consumption and preference. PMID- 15355329 TI - Mu-opioid receptor knockout mice show diminished food-anticipatory activity. AB - We have previously suggested that during or prior to activation of anticipatory behaviour to a coming reward, mu-opioid receptors are activated. To test this hypothesis schedule induced food-anticipatory activity in mu-opioid receptor knockout mice was measured using running wheels. We hypothesized that mu-knockout mice show little food-anticipatory activity. In wildtype mice we observed that food-anticipatory activity increased proportional to reduced food intake levels during daily scheduled food access, and thus reflects the animal's physiological need for food. mu-Knockout mice do not adjust their schedule induced running wheel behaviour prior to and during feeding time in the same way as wildtype mice; rather than showing more running wheel activity before than during feeding, they showed an equal amount of activity before and during feeding. As food anticipatory activity is dependent on the mesolimbic dopamine system and mu opioid receptors regulate dopaminergic activity, these data suggest a change in the dopamine system's activity in mu-knockout mice. As we observed that mu knockout mice tended to show a stronger locomotor activity response than wildtype mice to the indirect dopamine agonist d-amphetamine, it appears that the dopaminergic system per se is intact and sensitive to activation. We found no differences in the expression of pro-opiomelanocortin, a precursor of endogenous endorphin, in the arcuate nucleus between mu-knockout mice and wildtype mice during restricted feeding, showing that the mu-opioid receptor does not regulate endogenous endorphin levels. These data overall suggest a role for mu-opioid receptors in adapting reward related behaviour to the requirements of the environment. PMID- 15355330 TI - Metabotropic glutamate receptor 5 localized in the limbic forebrain is critical for the development of morphine-induced rewarding effect in mice. AB - The aim of the present study was to clarify the role of the metabotropic glutamate 5 (mGlu5) receptor subtype in the development of rewarding effect induced by a prototypical mu-opioid receptor agonist morphine in the mouse. In the conditioned place preference paradigm, intracerebroventricular (i.c.v.) administration of a selective mGlu5 receptor antagonist, 2-methyl-6 (phenylethynyl)-pyridine (MPEP), attenuated the morphine-induced rewarding effects. Using immunoblot analysis, we confirmed that the increased level of protein kinase Cgamma (PKCgamma) isoform was observed in the limbic forebrain of ICR mice conditioned with morphine. Here we found for the first time that the treatment with MPEP significantly inhibited the up-regulation of PKCgamma isoform in the limbic forebrain of mice showing the significant place preference. Furthermore, it should be mentioned that the protein level of mGlu5 was significantly increased in membrane preparations of the limbic forebrain obtained from morphine-conditioned mice compared to those from saline-conditioned mice. As well as the result from the immunoblot analysis, we demonstrated using the receptor binding assay that the number of mGlu5 receptors in the mouse limbic forebrain was significantly increased by morphine conditioning. The present data provide direct evidence that the activation of mGlu5 receptor linked to the increased PKCgamma isoform in the mouse limbic forebrain is implicated in the development of rewarding effect of morphine. PMID- 15355331 TI - Cocaine increases medial prefrontal cortical glutamate overflow in cocaine sensitized rats: a time course study. AB - Excitatory amino acid transmission within mesocorticolimbic brain pathways is thought to play an important role in behavioural sensitization to psychomotor stimulants. The current studies evaluated a time course of the effects of cocaine on extracellular glutamate levels within the medial prefrontal cortex (mPFC) following increasing periods of withdrawal from repeated cocaine exposure. Male Sprague-Dawley rats underwent stereotaxic surgeries and were pretreated daily with saline (1 mL/kg/day x 4 days, i.p.) or cocaine (15 mg/kg/day x 4 days, i.p.) and withdrawn for 1, 7 or 30 days. After withdrawal rats were challenged with the same dose of saline or cocaine and in vivo microdialysis of the mPFC was conducted with concurrent analysis of locomotor activity. Animals that were withdrawn from repeated daily cocaine for 1 day and 7 days displayed an augmentation in cocaine-induced mPFC glutamate levels compared to saline and acute control subjects, which were similarly unaffected by cocaine challenge. At the 7 day time point, a subset of animals that received repeated cocaine did not express behavioural sensitization, nor did these animals exhibit the enhancement in mPFC glutamate in response to cocaine challenge. In contrast to these early effects, 30 days of withdrawal resulted in no significant changes in cocaine induced mPFC glutamate levels regardless of the pretreatment or behavioural response. These data suggest that repeated cocaine administration transiently increases cocaine-induced glutamate levels in the mPFC during the first week of withdrawal, which may play an important role in the development of behavioural sensitization to cocaine. PMID- 15355332 TI - The induction of behavioural sensitization is associated with cocaine-induced structural plasticity in the core (but not shell) of the nucleus accumbens. AB - Repeated exposure to cocaine increases the density of dendritic spines on medium spiny neurons in the nucleus accumbens (Acb) and pyramidal cells in the medial prefrontal cortex (mPFC). To determine if this is associated with the development of psychomotor sensitization, rats were given daily i.p. injections of 15 mg/kg of cocaine (or saline) for 8 days, either in their home cage (which failed to induce significant psychomotor sensitization) or in a distinct and relatively novel test cage (which induced robust psychomotor sensitization). Their brains were obtained 2 weeks after the last injection and processed for Golgi-Cox staining. In the Acb core (AcbC) cocaine treatment increased spine density only in the group that developed psychomotor sensitization (i.e. in the Novel but not Home group), and there was a significant positive correlation between the degree of psychomotor sensitization and spine density. In the Acb shell (AcbS) cocaine increased spine density to the same extent in both groups; i.e. independent of psychomotor sensitization. In the mPFC cocaine increased spine density in both groups, but to a significantly greater extent in the Novel group. Furthermore, when rats were treated at Home with a higher dose of cocaine (30 mg/kg), cocaine now induced psychomotor sensitization in this context, and also increased spine density in the AcbC. Thus, the context in which cocaine is experienced influences its ability to reorganize patterns of synaptic connectivity in the Acb and mPFC, and the induction of psychomotor sensitization is associated with structural plasticity in the AcbC and mPFC, but not the AcbS. PMID- 15355333 TI - Beneficial effects of enriched environment on adolescent rats from stressed pregnancies. AB - The capacity of an early environmental intervention to normalize the behavioural and immunological dysfunctions produced by a stressed pregnancy was investigated. Pregnant Sprague-Dawley rats underwent three 45-min sessions per day of prenatal restraint stress (PS) on gestation days 11-21, and their offspring were assigned to either an enriched-environment or standard living cages throughout adolescence [postnatal days (pnd) 22-43]. Juvenile rats from stressed pregnancies had a prominent depression of affiliative/playful behaviour and of basal circulating CD4 T lymphocytes, CD8 T lymphocytes and T4/T8 ratio. They also showed increased emotionality and spleen and brain frontal cortex levels of pro-inflammatory interleoukin-1beta (IL-1beta) cytokine. A more marked response to cyclophosphamide (CPA: two 2 mg/kg IP injections) induced immunosuppression was also found in prenatal stressed rats. Enriched housing increased the amount of time adolescent PS rats spent in positive species-typical behaviours (i.e. play behaviour), reduced emotionality and reverted most of immunological alterations. In addition to its effects in PS rats, enriched housing increased anti inflammatory IL-2 and reduced pro-inflammatory IL-1beta production by activated splenocytes, also producing a marked alleviation of CPA-induced immune depression. In the brain, enriched housing increased IL-1beta values in hypothalamus, while slightly normalizing these values in the frontal cortex from PS rats. This is a first indication that an environmental intervention, such as enriched housing, during adolescence can beneficially affect basal immune parameters and rats response to both early stress and drug-induced immunosuppression. PMID- 15355334 TI - Chronic stress attenuates GABAergic inhibition and alters gene expression of parvocellular neurons in rat hypothalamus. AB - Chronic stress causes disinhibition of the hypothalamus-pituitary-adrenal axis. Consequently, the brain is overexposed to glucocorticoids which in humans may precipitate stress-related disorders, e.g. depression. The hypothalamus-pituitary adrenal activity is strongly regulated by GABAergic input to parvocellular neurons in the hypothalamic paraventricular nucleus. We here report a reduced frequency of miniature inhibitory postsynaptic currents (mIPSCs) in parvocellular neurons of rats exposed to 3 weeks of unpredictable stress. The mIPSC amplitude and kinetic properties were unchanged, pointing to a presynaptic change caused by chronic stress. Because paired-pulse inhibition was unaffected by chronic stress, the number of functional GABAergic synaptic contacts rather than the release probability seems to be reduced after chronic stress. Linearly amplified RNA from postsynaptic cells was hybridized with multiple cDNA clones of interest, including most GABA(A) receptor subunits. In agreement with the electrophysiological observations, relative expression of the prevalent GABA(A)alpha1, alpha3, gamma1 and gamma2 receptor subunits, which largely contribute to the recorded responses, was not altered after chronic stress. However, expression of the extra-synaptic GABA(A)alpha5 subunit, earlier linked to depression in humans, and of the delta receptor subunit were found to be significantly changed. In conclusion, chronic stress leads to presynaptic functional alterations in GABAergic input to the paraventricular nucleus which could contribute to the observed disinhibition of the hypothalamus-pituitary adrenal axis; additionally other aspects of GABAergic transmission may also be changed due to transcriptional regulation of specific receptor subunits in the parvocellular neurons. PMID- 15355335 TI - Microstimulation of V1 input layers disrupts the selection and detection of visual targets by monkeys. AB - Electrical microstimulation delivered to primary visual cortex (V1) concurrently with the presentation of visual targets interferes with the selection of these targets. To determine the source of this interference, we stimulated the visual input layers of V1 as rhesus monkeys generated saccadic eye movements to visual targets presented at and outside the receptive field of the stimulated neurons. Columns of cells in V1 innervated by the left and right eye are segregated according to eye dominance, such that cells within a column respond best to visual stimuli presented to the ocular dominant eye. Interference was maximal when targets were presented to the ocular dominant eye, moderate when presented to the ocular inferior eye, and negligible when presented to both eyes. Thus, electrical microstimulation of the visual input layers of V1 disrupts the flow of visual information along the geniculostriate pathway. Knowing how electrical stimulation of V1 affects visual behaviour is necessary when using monkeys to develop a visual prosthesis for the blind. PMID- 15355336 TI - Specific and nonspecific effects of transcranial magnetic stimulation on picture word verification. AB - Repetitive transcranial magnetic stimulation (rTMS) can temporarily impair or improve performance, including language processing. It remains unclear, however, (i) which scalp sites are most appropriate to achieve the desired effects and (ii) which experimental setups produce facilitation or inhibition of language functions. We assessed the effects of TMS at different stimulation sites on picture-word verification in healthy volunteers. Twenty healthy volunteers with left language lateralization, as determined by functional transcranial Dopplersonography, performed picture-word verification prior to and after rTMS (1 Hz for 600 s at 110% of subjects' resting motor thresholds). Stimulation sites were the classical language areas (Broca's and Wernicke's), their homolog brain regions of the right hemisphere, and the occipital cortex. Additionally, sham stimulation over Broca's area was applied in a subsample of 11 subjects. As a control task, 10 volunteers performed a colour-tone matching task under the same experimental conditions. There was a general nonspecific arousal effect for both verum and sham TMS for both the picture-word verification and for the control task. However, superimposed there were opposite effects on picture-word verification for stimulation of Wernicke's area and Broca's area, namely a relative inhibition in the case of Wernicke's area and a relative facilitation in the case of Broca's area. These results demonstrate that low frequency rTMS has both general arousing effects and domain-specific effects. PMID- 15355337 TI - Associative somatodendritic interaction in layer V pyramidal neurons is not affected by the antiepileptic drug lamotrigine. AB - The antiepileptic drug lamotrigine was described to exert its effects on neuronal excitability via voltage-gated sodium and calcium, as well as hyperpolarization activated conductances. In order to define the effects of lamotrigine on the excitability of layer V pyramidal cells of the rat somatosensory cortex we performed patch-clamp recordings from the soma and dendrite of this major cortical output cell type in acute slices. Voltage-clamp experiments revealed the blockade of the persistent sodium current by 50-100 micro m lamotrigine as well as by 50 micro m of the anticonvulsant drug phenytoin. In somatic current-clamp studies lamotrigine, in a therapeutic concentration range, depolarizes the membrane potential reflecting the activation of the hyperpolarization-activated current. This depolarization reduces the rheobase and increases the spiking frequency at the onset of the spike train. For long depolarizing current pulses under lamotrigine, however, a use-dependent block of sodium channels reduces spiking frequency and spike amplitude. The depolarization due to 50-100 micro m lamotrigine reduces additionally the critical frequency of back-propagating spikes necessary to elicit a dendritic calcium action potential. Ten to thirty micromolar lamotrigine, in contrast, did not change the critical frequency. Lamotrigine blocks long-lasting, high frequent spiking activity due to its use dependent sodium channel block, while burst activity is not impaired due to a depolarizing shift of the membrane potential. This drug therefore dampens epileptic activity while leaving the somatodendritic association in layer V pyramidal cells intact. PMID- 15355339 TI - Ras oncogene induces beta-galactoside alpha2,6-sialyltransferase (ST6Gal I) via a RalGEF-mediated signal to its housekeeping promoter. AB - Several oncogenic proteins are known to influence cellular glycosylation. In particular, transfection of codon 12 point mutated H-Ras increases CMP-Neu5Ac: Galbeta1,4GlcNAc alpha2,6-sialyltransferase I (ST6Gal I) activity in rodent fibroblasts. Given that Ras mediates its effects through at least three secondary effector pathways (Raf, RalGEFs and PI3K) and that transcriptional control of mouse ST6Gal I is achieved by the selective use of multiple promoters, we attempted to identify which of these parameters are involved in linking the Ras signal to ST6Gal I gene transcription in mouse fibroblasts. Transformation by human K-Ras or H-Ras (S12 and V12 point mutations, respectively) results in a 10 fold increase in ST6Gal I mRNA, but no alteration in the expression of related sialyltransferases. Using an inducible H-RasV12 expression system, a direct causal link between activated H-Ras expression and elevated ST6Gal I mRNA was demonstrated. The accumulation of the ST6Gal I transcript in response to activated Ras was accompanied by an increase of alpha2,6-sialyltransferase activity and of Neu5Acalpha2,6Gal at the cell surface. Results obtained with H RasV12 partial loss of function mutants H-RasV12S35 (Raf signal only), H RasV12C40 (PI3-kinase signal only) and H-RasV12G37 (RalGEFs signal only) suggest that the H-Ras induction of the mouse ST6Gal I gene (Siat1) transcription is primarily routed through RalGEFs. 5'-Rapid amplification of cDNA ends analysis demonstrated that the increase in ST6Gal I mRNA upon H-RasV12 or K-RasS12 transfection is mediated by the Siat1 housekeeping promoter P3-associated 5' untranslated exons. PMID- 15355340 TI - Characterization of recombinant forms of the yeast Gas1 protein and identification of residues essential for glucanosyltransferase activity and folding. AB - Gas1p is a glycosylphosphatidylinositol-anchored plasma membrane glycoprotein of Saccharomyces cerevisiae and is a representative of Family GH72 of glycosidases/transglycosidases, which also includes proteins from human fungal pathogens. Gas1p, Phr1-2p from Candida albicans and Gel1p from Aspergillus fumigatus have been shown to be beta-(1,3)-glucanosyltransferases required for proper cell wall assembly and morphogenesis. Gas1p is organized into three modules: a catalytic domain; a cys-rich domain; and a highly O-glycosylated serine-rich region. In order to provide an experimental system for the biochemical and structural analysis of Gas1p, we expressed soluble forms in the methylotrophic yeast Pichia pastoris. Here we report that 48 h after induction with methanol, soluble Gas1p was produced at a yield of approximately 10 mg x L( 1) of medium, and this value was unaffected by the further removal of the serine rich region or by fusion to a 6 x His tag. Purified soluble Gas1 protein showed beta-(1,3)-glucanosyltransferase activity that was abolished by replacement of the putative catalytic residues, E161 and E262, with glutamine. Spectral studies confirmed that the recombinant soluble Gas1 protein assumed a stable conformation in P. pastoris. Interestingly, thermal denaturation studies demonstrated that Gas1p is highly resistant to heat denaturation, and a complete refolding of the protein following heat treatment was observed. We also showed that Gas1p contains five intrachain disulphide bonds. The effects of the C74S, C103S and C265S substitutions in the membrane-bound Gas1p were analyzed in S. cerevisiae. The Gas1-C74S protein was totally unable to complement the phenotype of the gas1 null mutant. We found that C74 is an essential residue for the proper folding and maturation of Gas1p. PMID- 15355341 TI - Antioxidant defences and homeostasis of reactive oxygen species in different human mitochondrial DNA-depleted cell lines. AB - Three pairs of parental (rho+) and established mitochondrial DNA depleted (rho0) cells, derived from bone, lung and muscle were used to verify the influence of the nuclear background and the lack of efficient mitochondrial respiratory chain on antioxidant defences and homeostasis of intracellular reactive oxygen species (ROS). Mitochondrial DNA depletion significantly lowered glutathione reductase activity, glutathione (GSH) content, and consistently altered the GSH2 : oxidized glutathione ratio in all of the rho0 cell lines, albeit to differing extents, indicating the most oxidized redox state in bone rho0 cells. Activity, as well as gene expression and protein content, of superoxide dismutase showed a decrease in bone and muscle rho0 cell lines but not in lung rho0 cells. GSH peroxidase activity was four times higher in all three rho0 cell lines in comparison to the parental rho+, suggesting that this may be a necessary adaptation for survival without a functional respiratory chain. Taken together, these data suggest that the lack of respiratory chain prompts the cells to reduce their need for antioxidant defences in a tissue-specific manner, exposing them to a major risk of oxidative injury. In fact bone-derived rho0 cells displayed the highest steady state level of intracellular ROS (measured directly by 2',7'-dichlorofluorescin, or indirectly by aconitase activity) compared to all the other rho+ and rho0 cells, both in the presence or absence of glucose. Analysis of mitochondrial and cytosolic/iron regulatory protein-1 aconitase indicated that most ROS of bone rho0 cells originate from sources other than mitochondria. PMID- 15355342 TI - Evidence for the presence of ferritin in plant mitochondria. AB - In this work, evidence for the presence of ferritins in plant mitochondria is supplied. Mitochondria were isolated from etiolated pea stems and Arabidopsis thaliana cell cultures. The proteins were separated by SDS/PAGE. A protein, with an apparent molecular mass of approximately 25-26 kDa (corresponding to that of ferritin), was cross-reacted with an antibody raised against pea seed ferritin. The mitochondrial ferritin from pea stems was also purified by immunoprecipitation. The purified protein was analyzed by MALDI-TOF mass spectrometry and the results of both mass finger print and peptide fragmentation by post source decay assign the polypeptide sequence to the pea ferritin (P < 0.05). The mitochondrial localization of ferritin was also confirmed by immunocytochemistry experiments on isolated mitochondria and cross-sections of pea stem cells. The possible role of ferritin in oxidative stress of plant mitochondria is discussed. PMID- 15355343 TI - Determination of the reopening temperature of a DNA hairpin structure in vitro. AB - A novel method, based upon primer extension, has been developed for measuring the reopening temperature of a single type of DNA hairpin structure. Two DNA oligonucleotides have been utilized and designated as primers 1 and 2. Primer 1, with its 5- and 3'-termini fully complementary to the hairpin flanking sequences, was used to evaluate primer extension conditions, and primer 2, with its 3'-end competing with the DNA hairpin stem, was used to detect the DNA hairpin reopening temperature. A single DNA hairpin structure was formed on the DNA template by thermal denaturation and renaturation, and this hairpin structure was predicted to prevent the annealing of the 3'-end of primer 2 with the template DNA, which leads to no primer extension. By incubating at different temperatures, the DNA hairpin structure can be reopened at a particular temperature where the primer extension can be carried out. This resulted in the appearance of double-stranded DNA that was detected on an agarose gel. This temperature is defined here as the hairpin reopening temperature. PMID- 15355344 TI - Biosynthesis of platelet glycoprotein V expressed as a single subunit or in association with GPIb-IX. AB - Glycoprotein (GP) V is noncovalently linked to GPIbalpha, GPIbbeta and GPIX within the platelet GPIb-V-IX complex, a receptor for von Willebrand factor and thrombin. Two functions have been ascribed to GPV, namely, the modulation of thrombin- and collagen-dependent platelet responses. The biosynthesis of this molecule was investigated in pulse-chase metabolic labelling experiments performed in CHO cell lines transfected with GPV, alone or in the presence of GPIb-IX. GPV could not be detected at the surface of cells expressing the single subunit but was found instead as a soluble form in the culture medium. In pulse chase studies, an immature 70 kDa protein was detected in cell lysates, whereas a fully processed 80-82 kDa form was only observed in the culture supernatants at later chase times. Immature GPV was N-glycosylated and retained before the medial Golgi while the secreted molecule contained complex sialylated sugars. The mature soluble form of GPV was produced by an enzymatic cleavage which was not affected by inhibitors of proteasome, calpain or metalloproteinases. When GPV was cotransfected with GPIb-IX, the former was no longer found in the culture supernatant but was retained in the cell membrane as shown by fluorescence activated cell sorting and confocal microscopy analyses. Surface expressed GPV was processed from an immature 70 kDa form to produce a mature 80 kDa protein, processing similar to the intracellular trafficking of GPIbalpha. These results indicate that correct biosynthesis and surface expression of GPV in platelets requires the presence of the other subunits of the GPIb-V-IX complex. PMID- 15355345 TI - l-3,4-Dihydroxyphenyl alanine-extradiol cleavage is followed by intramolecular cyclization in lincomycin biosynthesis. AB - The LmbB1 protein, participating in the biosynthesis of lincomycin, was heterologously expressed in Escherichia coli, purified in its active form, and characterized as a dimer of identical subunits. Methods for purification and analysis of the LmbB1 reaction product were developed. Molecular mass and fragmentation pattern of the product revealed by capillary electrophoresis-mass spectrometry were in agreement with its proposed structure, 4-(3-carboxy-3-oxo propenyl)-2,3-dihydro-1H-pyrrole-2-carboxylic acid. The LmbB1 is therefore a dioxygenase catalysing the 2,3-extradiol cleavage of the l-3,4-dihydroxyphenyl alanine aromatic ring. The final LmbB1 reaction product, a unique compound found in biosynthesis of lincomycin and expected in anthramycins, arises through subsequent cyclization of the primary cleavage product, 2,3-secodopa. A possible role of LmbB1 in 2,3-secodopa cyclization and alternative ways of the cyclization in the formation of biosynthetically related compounds, muscaflavin and stizolobinic acid, are discussed. PMID- 15355346 TI - Chaperone activity of recombinant maize chloroplast protein synthesis elongation factor, EF-Tu. AB - The protein synthesis elongation factor, EF-Tu, is a protein that carries aminoacyl-tRNA to the A-site of the ribosome during the elongation phase of protein synthesis. In maize (Zea mays L) this protein has been implicated in heat tolerance, and it has been hypothesized that EF-Tu confers heat tolerance by acting as a molecular chaperone and protecting heat-labile proteins from thermal aggregation and inactivation. In this study we investigated the effect of the recombinant precursor of maize EF-Tu (pre-EF-Tu) on thermal aggregation and inactivation of the heat-labile proteins, citrate synthase and malate dehydrogenase. The recombinant pre-EF-Tu was purified from Escherichia coli expressing this protein, and mass spectrometry confirmed that the isolated protein was indeed maize EF-Tu. The purified protein was capable of binding GDP (indicative of protein activity) and was stable at 45 degrees C, the highest temperature used in this study to test this protein for possible chaperone activity. Importantly, the recombinant maize pre-EF-Tu displayed chaperone activity. It protected citrate synthase and malate dehydrogenase from thermal aggregation and inactivation. To our knowledge, this is the first observation of chaperone activity by a plant/eukaryotic pre-EF-Tu protein. The results of this study support the hypothesis that maize EF-Tu plays a role in heat tolerance by acting as a molecular chaperone and protecting chloroplast proteins from thermal aggregation and inactivation. PMID- 15355347 TI - Mechanism for transcriptional synergy between interferon regulatory factor (IRF) 3 and IRF-7 in activation of the interferon-beta gene promoter. AB - The interferon-beta promoter has been studied extensively as a model system for combinatorial transcriptional regulation. In virus-infected cells the transcription factors ATF-2, c-Jun, interferon regulatory factor (IRF)-3, IRF-7 and NF-kappaB, and the coactivators p300/CBP play critical roles in the activation of this and other promoters. It remains unclear, however, why most other combinations of AP-1, IRF and Rel proteins fail to activate the interferon beta gene. Here we have explored how different IRFs may cooperate with other factors to activate transcription. First we showed in undifferentiated embryonic carcinoma cells that ectopic expression of either IRF-3 or IRF-7, but not IRF-1, was sufficient to allow virus-dependent activation of the interferon-beta promoter. Moreover, the activity of IRF-3 and IRF-7 was strongly affected by promoter context, with IRF-7 preferentially being recruited to the natural interferon-beta promoter. We fully reconstituted activation of this promoter in insect cells. Maximal synergy required IRF-3 and IRF-7 but not IRF-1, and was strongly dependent on the presence of p300/CBP, even when these coactivators only modestly affected the activity of each factor by itself. These results suggest that specificity in activation of the interferon-beta gene depends on a unique promoter context and on the role played by coactivators as architectural factors. PMID- 15355348 TI - Differences in substrate specificities between cysteine protease CPB isoforms of Leishmania mexicana are mediated by a few amino acid changes. AB - The CPB genes of the protozoan parasite Leishmania mexicana encode stage regulated cathepsin L-like cysteine proteases that are important virulence factors and are in a tandem array of 19 genes. In this study, we have compared the substrate preferences of two CPB isoforms, CPB2.8 and CPB3, and a H84Y mutant of the latter enzyme, to analyse the roles played by the few amino acid differences between the isoenzymes in determining substrate specificity. CPB3 differs from CPB2.8 at just three residues (N60D, D61N and D64S) in the mature domain. The H84Y mutation mimics an additional change present in another isoenzyme, CPB18. The active recombinant CPB isoenzymes and mutant were produced using Escherichia coli and the S1-S3 and S1'-S3' subsite specificities determined using a series of fluorogenic peptide derivatives in which substitutions were made on positions P3 to P3' by natural amino acids. Carboxydipeptidase activities of CPB3 and H84Y were also observed using the peptide Abz-FRAK(Dnp)-OH and some of its analogues. The kinetic parameters of hydrolysis by CPB3, H84Y and CPB2.8 of the synthetic substrates indicates that the specificity of S3 to S3' subsites is influenced greatly by the modifications at amino acids 60, 61, 64 and 84. Particularly noteworthy was the large preference for Pro in the P2' position for the hydrolytic activity of CPB3, which may be relevant to a role in the activation mechanism of the L. mexicana CPBs. PMID- 15355349 TI - The HNF1beta transcription factor has several domains involved in nephrogenesis and partially rescues Pax8/lim1-induced kidney malformations. AB - The tissue-specific transcription factors HNF1alpha and HNF1beta are closely related homeodomain proteins conserved in vertebrate evolution. Heterozygous mutations in human HNF1beta but not in HNF1alpha genes are associated with kidney malformations. Overexpression of HNF1beta in Xenopus embryos leads to defective pronephros development, while HNF1alpha has no effect. We have defined the regions responsible for this functional difference between HNF1beta and HNF1alpha in transfected HeLa cells as well as in injected Xenopus embryos. Using domain swapping experiments, we located a nuclear localization signal in the POUH domain of HNF1beta, and showed that the POUS and POUH domains of HNF1beta mediate a high transactivation potential in transfected cells. In injected Xenopus embryos three HNF1beta domains are involved in nephrogenesis. These include the dimerization domain, the 26 amino acid segment specific for splice variant A as well as the POUH domain. As HNF1beta together with Pax8 and lim1 constitute the earliest regulators in the pronephric anlage, it is possible that they cooperate during early nephrogenesis. We have shown here that HNF1beta can overcome the enlargement and the induction of an ectopic pronephros mediated by overexpression of Pax8 and lim1. However, the phenotype induced by Pax8 and lim1 overexpression and characterized by cyst-like structures and thickening of the pronephric tubules was not altered by HNF1beta overexpression. Taken together, HNF1beta acts antagonistically to Pax8 and lim1 in only some processes during nephrogenesis, and a simple antagonistic relationship does not completely describe the functions of these genes. We conclude that HNF1beta has some distinct morphogenetic properties during nephrogenesis. PMID- 15355350 TI - Integrin receptor specificity for human red cell ICAM-4 ligand. Critical residues for alphaIIbeta3 binding. AB - The red cell intercellular adhesion molecule-4 (ICAM-4) binds to different members of the integrin receptor families. To better define the ICAM-4 integrin receptor specificity, cell transfectants individually expressing various integrins were used to demonstrate that alphaLbeta2, alphaMbeta2, and alphaIIbbeta3 (activated) bind specifically and dose dependently to the recombinant ICAM-4-Fc protein. We also show that cell surface ICAM-4 interacts with the cell surface alphaVbeta3 integrin. In addition, using a alpha4beta1 cell transfectant and beta2 integrin-deficient LAD cells, we show here that ICAM-4 failed to interact with alpha4beta1 even after alpha4beta1 activation by phorbol ester or with the monoclonal antibody TS2/16 (+ Mn2+). ICAM-4 amino acids that are critical for alphaIIbbeta3 and alphaVbeta3 interaction were identified by domain deletion analysis, site-directed mutagenesis and synthetic peptide inhibition. Our results provide evidence that the beta3 integrin binding sites encompass the first and second Ig-like domains of ICAM-4. However, while the alphaIIbbeta3 contact site comprises the ABED face of domain D1 with an extension in the C'-E loop of domain D2, the alphaVbeta3 contact site comprises residues on both faces of D1 and in the C'-E loop of D2. These data, together with our previous results, demonstrate that different integrins bind to different but partly overlapping sites on ICAM-4, and that ICAM-4 may accommodate multiple integrin receptors present on leukocytes, platelets and endothelial cells. PMID- 15355351 TI - RNA helicase A interacts with nuclear factor kappaB p65 and functions as a transcriptional coactivator. AB - RNA helicase A (RHA), a member of DNA and RNA helicase family containing ATPase activity, is involved in many steps of gene expression such as transcription and mRNA export. RHA has been reported to bind directly to the transcriptional coactivator, CREB-binding protein, and the tumor suppressor protein, BRCA1, and links them to RNA Polymerase II holoenzyme complex. Using yeast two-hybrid screening, we have identified RHA as an interacting molecule of the p65 subunit of nuclear factor kappaB (NF-kappaB). The interaction between p65 and RHA was confirmed by glutathione-S transferase pull-down assay in vitro, and by co immunoprecipitation assay in vivo. In transient transfection assays, RHA enhanced NF-kappaB dependent reporter gene expression induced by p65, tumor necrosis factor-alpha, or NF-kappaB inducing kinase. The mutant form of RHA lacking ATP binding activity inhibited NF-kappaB dependent reporter gene expression induced by these activators. Moreover, depletion of RHA using short interfering RNA reduced the NF-kappaB dependent transactivation. These data suggest that RHA is an essential component of the transactivation complex by mediating the transcriptional activity of NF-kappaB. PMID- 15355352 TI - Expression in yeast of a novel phospholipase A1 cDNA from Arabidopsis thaliana. AB - During a search for cDNAs encoding plant sterol acyltransferases, we isolated four full-length cDNAs from Arabidopsis thaliana that encode proteins with substantial identity with animal lecithin : cholesterol acyltransferases (LCATs). The expression of one of these cDNAs, AtLCAT3 (At3g03310), in various yeast strains resulted in the doubling of the triacylglycerol content. Furthermore, a complete lipid analysis of the transformed wild-type yeast showed that its phospholipid content was lower than that of the control (void plasmid transformed) yeast whereas lysophospholipids and free fatty acids increased. When microsomes from the AtLCAT3-transformed yeast were incubated with di-[1-14C]oleyl phosphatidylcholine, both the lysophospholipid and free fatty acid fractions were highly and similarly labelled, whereas the same incubation with microsomes from the control yeast produced a negligible labelling of these fractions. Moreover when microsomes from AtLCAT3-transformed yeast were incubated with either sn-1- or sn-2-[1-14C]acyl phosphatidylcholine, the distribution of the labelling between the free fatty acid and the lysophosphatidylcholine fractions strongly suggested a phospholipase A1 activity for AtLCAT3. The sn-1 specificity of this phospholipase was confirmed by gas chromatography analysis of the hydrolysis of 1 myristoyl, 2-oleyl phosphatidylcholine. Phosphatidylethanolamine and phosphatidic acid were shown to be also hydrolysed by AtLCAT3, although less efficiently than phosphatidylcholine. Lysophospatidylcholine was a weak substrate whereas tripalmitoylglycerol and cholesteryl oleate were not hydrolysed at all. This novel A. thaliana phospholipase A1 shows optimal activity at pH 6-6.5 and 60-65 degrees C and appears to be unaffected by Ca2+. Its sequence is unrelated to all other known phospholipases. Further studies are in progress to elucidate its physiological role. PMID- 15355353 TI - Identification of intracellular target proteins of the calcium-signaling protein S100A12. AB - In this report, we have focused our attention on identifying intracellular mammalian proteins that bind S100A12 in a Ca2+-dependent manner. Using S100A12 affinity chromatography, we have identified cytosolic NADP+-dependent isocitrate dehydrogenase (IDH), fructose-1,6-bisphosphate aldolase A (aldolase), glyceraldehyde-3-phosphate dehydrogenese (GAPDH), annexin V, S100A9, and S100A12 itself as S100A12-binding proteins. Immunoprecipitation experiments indicated the formation of stable complexes between S100A12 and IDH, aldolase, GAPDH, annexin V and S100A9 in vivo. Surface plasmon resonance analysis showed that the binding to S100A12, of S100A12, S100A9 and annexin V, was strictly Ca2+-dependent, whereas that of GAPDH and IDH was only weakly Ca2+-dependent. To localize the site of S100A12 interaction, we examined the binding of a series of C-terminal truncation mutants to the S100A12-immobilized sensor chip. The results indicated that the S100A12-binding site on S100A12 itself is located at the C-terminus (residues 87 92). However, cross-linking experiments with the truncation mutants indicated that residues 87-92 were not essential for S100A12 dimerization. Thus, the interaction between S100A12 and S100A9 or immobilized S100A12 should not be viewed as a typical S100 homo- or heterodimerization model. Ca2+-dependent affinity chromatography revealed that C-terminal residues 75-92 are not necessary for the interaction of S100A12 with IDH, aldolase, GAPDH and annexin V. To analyze the functional properties of S100A12, we studied its action in protein folding reactions in vitro. The thermal aggregation of IDH or GAPDH was facilitated by S100A12 in the absence of Ca2+, whereas in the presence of Ca2+ the protein suppressed the aggregation of aldolase to less than 50%. These results suggest that S100A12 may have a chaperone/antichaperone-like function which is Ca2+-dependent. PMID- 15355354 TI - Structured DNA promotes phosphorylation of p53 by DNA-dependent protein kinase at serine 9 and threonine 18. AB - Phosphorylation at multiple sites within the N-terminus of p53 promotes its dissociation from hdm2/mdm2 and stimulates its transcriptional regulatory potential. The large phosphoinositide 3-kinase-like kinases ataxia telangiectasia mutated gene product and the ataxia telangectasia and RAD-3-related kinase promote phosphorylation of human p53 at Ser15 and Ser20, and are required for the activation of p53 following DNA damage. DNA-dependent protein kinase (DNA-PK) is another large phosphoinositide 3-kinase-like kinase with the potential to phosphorylate p53 at Ser15, and has been proposed to enhance phosphorylation of these sites in vivo. Moreover, recent studies support a role for DNA-PK in the regulation of p53-mediated apoptosis. We have shown previously that colocalization of p53 and DNA-PK to structured single-stranded DNA dramatically enhances the potential for p53 phosphorylation by DNA-PK. We report here the identification of p53 phosphorylation at two novel sites for DNA-PK, Thr18 and Ser9. Colocalization of p53 and DNA-PK on structured DNA was required for efficient phosphorylation of p53 at multiple sites, while specific recognition of Ser9 and Thr18 appeared to be dependent upon additional determinants of p53 beyond the N-terminal 65 amino acids. Our results suggest a role for DNA-PK in the modulation of p53 activity resultant from the convergence of p53 and DNA-PK on structured DNA. PMID- 15355355 TI - Irregular dimerization of guanylate cyclase-activating protein 1 mutants causes loss of target activation. AB - Guanylate cyclase-activating proteins (GCAPs) are neuronal calcium sensors that activate membrane bound guanylate cyclases (EC 4.6.1.2.) of vertebrate photoreceptor cells when cytoplasmic Ca2+ decreases during illumination. GCAPs contain four EF-hand Ca2+-binding motifs, but the first EF-hand is nonfunctional. It was concluded that for GCAP-2, the loss of Ca2+-binding ability of EF-hand 1 resulted in a region that is crucial for targeting guanylate cyclase [Ermilov, A.N., Olshevskaya, E.V. & Dizhoor, A.M. (2001) J. Biol. Chem.276, 48143-48148]. In this study we tested the consequences of mutations in EF-hand 1 of GCAP-1 with respect to Ca2+ binding, Ca2+-induced conformational changes and target activation. When the nonfunctional first EF-hand in GCAP-1 is replaced by a functional EF-hand the chimeric mutant CaM-GCAP-1 bound four Ca2+ and showed similar Ca2+-dependent changes in tryptophan fluorescence as the wild-type. CaM GCAP-1 neither activated nor interacted with guanylate cyclase. Size exclusion chromatography revealed that the mutant tended to form inactive dimers instead of active monomers like the wild-type. Critical amino acids in EF-hand 1 of GCAP-1 are cysteine at position 29 and proline at position 30, as changing these to glycine was sufficient to cause loss of target activation without a loss of Ca2+ induced conformational changes. The latter mutation also promoted dimerization of the protein. Our results show that EF-hand 1 in wild-type GCAP-1 is critical for providing the correct conformation for target activation. PMID- 15355356 TI - Escherichia coli cyclophilin B binds a highly distorted form of trans-prolyl peptide isomer. AB - Cyclophilins facilitate the peptidyl-prolyl isomerization of a trans-isomer to a cis-isomer in the refolding process of unfolded proteins to recover the natural folding state with cis-proline conformation. To date, only short peptides with a cis-form proline have been observed in complexes of human and Escherichia coli proteins of cyclophilin A, which is present in cytoplasm. The crystal structures analyzed in this study show two complexes in which peptides having a trans-form proline, i.e. succinyl-Ala-trans-Pro-Ala-p-nitroanilide and acetyl-Ala-Ala-trans Pro-Ala-amidomethylcoumarin, are bound on a K163T mutant of Escherichia coli cyclophilin B, the preprotein of which has a signal sequence. Comparison with cis form peptides bound to cyclophilin A reveals that in any case the proline ring is inserted into the hydrophobic pocket and a hydrogen bond between CO of Pro and Neta2 of Arg is formed to fix the peptide. On the other hand, in the cis-isomer, the formation of two hydrogen bonds of NH and CO of Ala preceding Pro with the protein fixes the peptide, whereas in the trans-isomer formation of a hydrogen bond between CO preceding Ala-Pro and His47 Nepsilon2 via a mediating water molecule allows the large distortion in the orientation of Ala of Ala-Pro. Although loss of double bond character of the amide bond of Ala-Pro is essential to the isomerization pathway occurring by rotating around its bond, these peptides have forms impossible to undergo proton transfer from the guanidyl group of Arg to the prolyl N atom, which induces loss of double bond character. PMID- 15355357 TI - Combination surgical lifting with ablative laser skin resurfacing of facial skin: a retrospective analysis. AB - BACKGROUND: Cutaneous aging is manifested by rhytides, dyschromias, and skin laxity. Ablative laser skin resurfacing can effectively improve many signs of skin aging; however, the photoaged patient with facial laxity often requires a surgical lifting procedure in order to obtain optimal results. Concerns with delayed or impaired wound healing has led to reluctance to perform both procedures simultaneously. OBJECTIVE: To report the clinical results and side effect profiles after concomitant surgical facial lifting procedures and ablative carbon dioxide or erbium:YAG laser resurfacing in a series of patients. METHODS: A retrospective analysis and chart review was performed in 34 consecutive patients who underwent combination CO(2) or erbium:YAG laser skin resurfacing and surgical lifting procedures, including S-lift rhytidectomy, blepharoplasty, and brow lift. Side effects and complication rates were tabulated. RESULTS: The side effect profile of the combined surgical-laser procedures was similar to that reported after a laser-only procedure. The most common side effect was transient hyperpigmentation which occurred in 20.6% of treated patients. None of the patients experienced delayed reepithelialization, skin necrosis, or prolonged healing times. CONCLUSIONS: Concurrent laser skin resurfacing and surgical lifting of facial skin maximizes aesthetic results without increased incidence of adverse effects. Patients benefit from the consolidation of anesthesia and convalescent times as well as enhanced global clinical outcomes. PMID- 15355358 TI - Intense pulsed light for the treatment of refractory melasma in Asian persons. AB - BACKGROUND: Patients with dermal or mixed-type melasmas are often refractory to various treatments. Intense pulsed light has been used to treat melanocytic lesions with promising results. OBJECTIVE: The purpose of this study was to clarify the effectiveness of intense pulsed light for refractory melasma in Asian persons. METHODS: Seventeen patients were treated with intense pulsed light, during four sessions at 4-week intervals. The patients were also given 4% hydroquinone cream and broad-spectrum sunscreens to prevent and treat postinflammatory hyperpigmentation. Sixteen patients in the control group were treated with hydroquinone cream and sunscreens. The treatment efficacy was evaluated using reflectance spectrophotometer and patient satisfaction questionnaire. RESULTS: Patients in the intense pulsed light group achieved an average of 39.8% improvement in relative melanin index, compared to 11.6% improvement in the control group (p<0.05) at Week 16. Six (35%) patients in the intense pulsed light group had more than 50% improvement, compared to two (14%) patients in the control group. Two patients in the intense pulsed light group, however, experienced transient postinflammatory hyperpigmentation, and partial repigmentation was noted 24 weeks after the last treatment session. CONCLUSION: Intense pulsed light is a safe and effective treatment for refractory melasma in Asian persons, with minimal side effects. Further treatment sessions are required for maintenance therapy. PMID- 15355359 TI - Minocycline-induced hyperpigmentation treated with a 755-nm Q-switched alexandrite laser. AB - BACKGROUND: Cutaneous pigmentation associated with minocycline therapy is an unusual adverse effect for which few successful treatments have been described. The pigment changes may persist for years, despite cessation of therapy, and is often cosmetically disfiguring, causing significant embarrassment and psychological depression in those affected. Few safe and effective treatments have been described in the past; however, recent pigment-specific laser technology has shown promise in the treatment of this condition. OBJECTIVE: The objective was to describe a series of patients with minocycline-induced hyperpigmentation who were successfully treated with a 755-nm Q-switched alexandrite laser. METHODS: Six patients with minocycline-induced hyperpigmentation on the face or legs were treated with a Q-switched alexandrite laser on a bimonthly basis until pigmentation was eradicated. RESULTS: Cutaneous pigmentation resolved completely in all patients in an average of four laser sessions. Side effects were limited to transient purpura and mild desquamation without scarring or dyspigmentation. CONCLUSION: Minocycline-induced cutaneous pigmentation can be effectively cleared without risk of adverse sequelae by Q switched alexandrite (755-nm) laser irradiation. PMID- 15355360 TI - Diode laser hair removal in patients undergoing isotretinoin therapy. AB - BACKGROUND: It is generally believed that any intervention in skin while on isotretinoin or within 6 months after the treatment can lead to prolonged healing and scarring. OBJECTIVE: The objective was to evaluate the side effects of laser hair removal in patients undergoing isotretinoin treatment. METHODS: Seven female patients undergoing isotretinoin therapy for acne were treated with a diode laser with a wavelength of 810 nm, a contact-cooling device, and a variable pulse width of 50 to 1000 ms. All volunteers received first treatment in the axillary or bikini area at 21 J/cm(2) (80 W) and the second treatment was done in the same area at 24 J/cm(2) (90 W). Six volunteers received additional treatments of chin area with a fluence of 21 or 24 J/cm(2). All volunteers were evaluated and photographed 1 week and 1 month after each treatment and degree of erythema, pigmentary change, vesiculation, swelling, and scarring was documented. RESULTS: There was no erythema, pigmentary change, swelling, or scarring at any follow up visits. One volunteer presented with a bulla 1 week after second treatment, which was resolved at 1-month follow-up. All volunteers were satisfied with the degree of hair removal with two treatments. CONCLUSION: This limited study suggests that diode laser hair removal is safe in patients undergoing isotretinoin therapy. PMID- 15355361 TI - Documenting dermatology practice: ratio of cutaneous tumors biopsied that are malignant. AB - BACKGROUND: Skin biopsy is an important part of dermatology practice. Little is known regarding dermatologist biopsying practices. OBJECTIVE: The objective was to assess the ratio of malignant lesions biopsied in practice and factors that influence this ratio. METHODS: Using a dermatopathology database, we analyzed tumors biopsied by dermatologists to determine the ratio of malignant to all tumors (malignancy ratio). Additional analyses were performed for factors influencing this ratio. RESULTS: Over 6 months, 11,072 tumors were submitted. Overall, the malignancy ratio was 41.7%, 3.8% for pigmented lesions. Practice type, location, or use of dermatoscopy did not influence this ratio; provider age was positively correlated with a higher malignancy ratio (r=0.51; p=0.02) and was a predictor of malignancy ratio (b=0.02; p=0.04). CONCLUSION: Approximately 40% of biopsied tumors were malignant. Dermatologists exhibit a wide range in their individual practice patterns. Increasing provider age predicted higher malignancy ratios with a 2% increase for each incremental year. PMID- 15355362 TI - Glove perforation in outpatient dermatologic surgery. AB - BACKGROUND: Intact surgical gloves ensure protection of health-care employees and health-care recipients. Nevertheless, glove perforation is very common and puncture rates above 50% have been published in the literature. OBJECTIVE: It was our aim to evaluate the perforation rate of surgical gloves in outpatient dermatologic surgery. METHODS: Six-hundred and sixty latex surgical gloves used in outpatient dermatologic surgery were evaluated for perforations using the approved water-leak method. Perforations were analyzed microscopically. RESULTS: Twenty of the 660 gloves were found to have perforations, which corresponds to a perforation rate of 3.0%. Only 5 of these perforations (25%) were noticed by the wearer. Perforations were more numerous in nondominant-handed gloves. Microscopically, all perforations could be identified as needle stick injuries. CONCLUSION: The risk of glove perforation in outpatient dermatologic surgery is lower than in many other medical specialties. The relatively high number of surgical procedures performed successively in outpatient dermatologic surgery, however, emphasizes the decisive relevance of an intact barrier between surgeon and patient. In view of the major fact that most perforations go unnoticed by the wearer, dermatologic surgeons must balance the improved safety of double gloving with costs and the loss of sensitivity and dexterity. PMID- 15355363 TI - Treatment of basal cell carcinoma with the pulsed carbon dioxide laser: a retrospective analysis. AB - BACKGROUND AND OBJECTIVE: Treatment options for basal cell carcinoma include surgical excision, cryotherapy, radiation, photodynamic therapy, Moh's micrographic surgery, and topical treatment with 5-fluorouracil and immunomodulators such as imiquimod. Resurfacing and ablation with a CO(2) laser (UltraPulse, Coherent Inc.) may present an attractive and effective treatment option in the management of these cutaneous cancers. We demonstrate the efficacy and safety of the UltraPulse CO(2) in the treatment of basal cell carcinomas of the skin. METHODS: We performed a retrospective chart review of 23 patients treated with the UltraPulse CO(2) laser. A total of 61 biopsy-proven superficial and nodular basal cell carcinomas without prior treatment were included in the study. The patients were followed postoperatively for a period of 15 to 85 months (mean 41.7 months) and assessed for clinical recurrence. RESULTS: Of the 61 tumors treated, clinical recurrence was observed in two cases (3.2%). Adverse effects included significant hypertrophic scarring in one patient and hypopigmentation in one patient. CONCLUSIONS: Destruction of superficial and nodular basal cell carcinomas may be accomplished successfully and safely with the UltraPulse CO(2) laser with a cure rate of 97%. PMID- 15355364 TI - The diameter of melanomas. AB - BACKGROUND: The mnemonic ABCD, which stands for asymmetry, border irregularity, color variation, and diameter greater than 6 mm alerts physicians to features that suggest melanoma. Although the ABCD criteria are guidelines for diagnosis, clinicians may overlook many melanomas that do not follow the ABCD rule. OBJECTIVE: We examined the sensitivity of the diameter portion of the ABCD rule. METHODS: Retrospective study examining the pathology reports of 383 melanomas. Data were compiled for each melanoma regarding its diameter, depth, body location, patient age, and sex. A 95% confidence interval (p=0.05) was used to identify the proportion of melanomas less than or equal to 6 mm in diameter. A two-tailed p value approach (p value =0.05, degrees of freedom=28) was used when evaluating two independent populations, lesions less than or equal to 6 mm and lesions greater than 6 mm in diameter. RESULTS: A total of 38.21% of melanomas were less than or equal to 6 mm in diameter after processing. Melanomas greater than 6 mm in diameter occurred in significantly older patients and at a greater Breslow's thickness than smaller melanomas. CONCLUSIONS: We demonstrated that a significant proportion of melanomas may be smaller than 6 mm. The ABCD criteria are not absolute; melanomas have many different appearances and start as small lesions. PMID- 15355365 TI - The rotating island pedicle flap: an aesthetic and functional improvement on the subcutaneous island pedicle flap. AB - BACKGROUND: The subcutaneous island pedicle flap is a useful closure in many areas on the face, particularly the cheek and upper lip. One disadvantage can be that the advancing edge of the flap leaves a scar at right angles to the local lines of relaxed skin tension. Tension on free edges such as the lid can also be a problem, but may be mitigated by incorporating an element of rotation into the closure. A variation on the subcutaneous island pedicle flap is described to help avoid the problems of the subcutaneous island pedicle flap while retaining its strengths. OBJECTIVE: Our experience with more than 120 rotating island pedicle flaps over 10 years is described to demonstrate the versatility of this subcutaneous island pedicle flap for facial and reconstructive surgery. METHODS: The technique is described and seven representative cases are presented with photographs. RESULTS: The flap is simple to perform, has minimal complications, and has good cosmetic results. Temporary alteration of sensation in the flap skin is a possible complication and can be largely avoided with a bipedicled dissection technique. CONCLUSION: Our experience shows the rotating island pedicle flap to be versatile, reliable, and aesthetic island pedicle flap for skin cancer surgery both on and off the face and a practical modification of the subcutaneous island pedicle flap. PMID- 15355366 TI - Dermoscopy of subcorneal hematoma. AB - BACKGROUND: Subcorneal hematoma is a pigmented skin lesion usually occurring on palms or soles after a trauma or sport activity. Clinically, it may exhibit overlapping features with acral melanoma or acral melanocytic nevi, leading to unnecessary excision of this otherwise harmless skin lesion. OBJECTIVE: The objective was to describe the dermoscopic features in a series of subcorneal hematomas. METHODS: Dermoscopic images of 15 subcorneal hematomas were evaluated for the presence of different colors and dermoscopic structures. RESULTS: In our series, a red-black hue was the most frequent color seen by dermoscopy (40% of the lesions) and a homogeneous pattern of pigmentation was the most frequent dermoscopic structure (53.3%). Remarkably, 40% of the lesions exhibited a parallel-ridge pattern that is usually found in early melanoma of palms and soles. In 46.7% of the lesions, red-black globules were additionally seen at the periphery as satellites disconnected from the lesion's body. Only two lesions showed either parallel-furrow or fibrillar pattern. A scratch test performed in four lesions, allowed complete or partial removal of the pigmentation. CONCLUSION: Dermoscopic features of subcorneal hematomas may be similar to those observed in acral melanocytic lesions. Nevertheless, in most cases the correct diagnosis can be facilitated by the presence of a red-black homogeneous pigmentation, often combined with satellite globules. A positive scratch test may be considered as an additional diagnostic clue. PMID- 15355367 TI - A case of foul genital odor treated with botulinum toxin A. AB - BACKGROUND: Genital odor is an uncommon condition characterized by an offensive and malodorous smell in the genital area. Although the etiology of foul genital odor is multifactorial, an important cause is sweat secretion and decomposition of sweat components by bacteria. Different methods are effective in reducing body odor secondary to bromhidrosis. Conservative methods only act for a short period of time, and more invasive surgical methods carry risk of complications or are inapplicable for the genital region. METHODS: A patient with localized foul odor in the genital hair bearing area was treated with botulinum toxin A. RESULTS: Botulinum toxin A was effective in creating an odorless and anhydrous response in the genital region, and no major adverse effects were noted during a follow-up of 9 months after injection. CONCLUSION: Local injection of botulinum toxin A appears to be a useful treatment for foul genital odor related to sweat glands activity. PMID- 15355368 TI - Use of a living dermal equivalent for a refractory abdominal defect after pediatric multivisceral transplantation. AB - BACKGROUND: Primary closure is not always possible after pediatric multivisceral transplantation. Reepithelialization may require extended periods of postoperative time, which can be associated with significant morbidity. OBJECTIVE: The objective was to accelerate secondary wound closure thereby minimizing infection or further complications in a pediatric multivisceral transplant patient. METHODS: Five applications of human fibroblast-derived dermis (Dermagraft, Smith and Nephew) were applied to the postsurgical defect of a pediatric multivisceral transplant patient over the course of 8 months. Routine wound care and observation was performed between human fibroblast-derived dermis applications. RESULTS: Human fibroblast-derived dermis stimulated healing and accelerated reepithelialization. Signs of clinical rejection or infection were not observed. CONCLUSION: Reepithelialization can be aided in the postoperative period in pediatric multivisceral transplant patients with human fibroblast derived dermis, thereby helping to deter complications associated with secondary wound closure. We have illustrated the successful use of a human fibroblast derived dermis as an adjunct for wound healing in a complicated surgical defect. PMID- 15355369 TI - Basal cell carcinoma arising in a port-wine stain. AB - BACKGROUND: The occurrence of basal cell carcinoma within a port-wine stain or nevus flammeus is rare. Sixteen cases of basal cell carcinoma which developed in a port-wine stain or nevus flammeus have been reported. OBJECTIVE: The objective was to demonstrate a rare case of basal cell carcinoma occurring in a port-wine stain successfully treated with Mohs micrographic surgery. METHODS: This is a case report and literature review. RESULTS: An 87-year-old man presented with a basal cell carcinoma on the margin of a previously untreated port-wine stain on the left cheek. Histologic examination showed a nodular basal cell carcinoma. The basal cell carcinoma was completely excised with Mohs micrographic surgery and complex linear closure was used to repair the wound in layers. The postoperative course was complicated by a hematoma, which developed 24 hr postoperatively. The hematoma was drained and there was no further bleeding or evidence of recurrence of the tumor after 12 months. CONCLUSION: Basal cell carcinoma should be included in the differential diagnosis of a skin lesion occurring in a port-wine stain. PMID- 15355370 TI - Multiple tumors of the follicular infundibulum. AB - BACKGROUND: Tumor of the follicular infundibulum is an uncommon benign neoplasm manifested histopathologically by a superficial epithelial plate-like growth pattern with multiple thin epidermal connections comprised of monomorphic cells with abundant cytoplasm. Cases of multiple tumors of the follicular infundibulum are rare and are described as hypopigmented scar-like macules or flat papules on the face, neck, and upper chest. OBJECTIVE: The objective was to describe a case of multiple tumors of the follicular infundibulum with numerous pigmented macules or papules and extensive involvement including the face, neck, anterior and posterior trunk, upper extremities, and intertriginous areas. METHODS: A case report and literature review are presented. CONCLUSION: Tumor of the follicular infundibulum with its characteristic histopathologic manifestations is a well recognized entity nowadays. Our case further expands the constellation of the clinical presentation of the multiple variant. Although the possibility of malignant basocellular degeneration seems remote, the multiplicity of the lesions, the possibility of clinical overlook, and the impracticality of complete treatment makes regular follow-up rational. PMID- 15355371 TI - Cutaneous leiomyoma with cytologic atypia, akin to uterine symplastic leiomyoma. AB - BACKGROUND: Smooth muscle tumors of the skin with cytologic pleomorphism and mitotic activity have traditionally been characterized as leiomyosarcomas, despite having a benign clinical course. In the uterus, there is a smooth muscle tumor known as symplastic leiomyoma, which typically has cytologic pleomorphism without significant mitotic activity. OBJECTIVE: The objective was to illustrate by case report the unusual histologic variant of the cutaneous pilar leiomyoma analogous to the symplastic leiomyoma of the uterus. METHODS: A 54-year-old woman presented with a clinical picture of cutaneous leiomyoma but had histologic features of nuclear hyperchromasia and pleomorphism, but rare mitoses. RESULTS: Management of this patient included excision of the involved area. CONCLUSION: Symplastic leiomyoma is an atypical uterine leiomyoma with cytologic atypia. We present the case of a patient with cutaneous leiomyomas that histologically demonstrated similar features to a uterine symplastic leiomyoma. We believe that this represents a distinct histologic variant of the cutaneous pilar leiomyoma analogous to the symplastic leiomyoma of the uterus. PMID- 15355372 TI - Vitamin B12-associated localized scleroderma and its treatment. AB - BACKGROUND: Localized drug-induced injection site reactions are rare. Injection site sclerodermoid reactions have been reported as a result of multiple drugs, most commonly vitamin K. Sclerodermoid injection site reactions have also been reported with pentazocine, progestin, and vitamin B12. Little has been written on the efficacious treatment of this complication. OBJECTIVE: The objective was to raise awareness of the potential complication of localized sclerodermoid change secondary to vitamin B12 injections and to present a novel approach to its treatment. METHODS: We report a case and review the English literature for injection site reactions from vitamin B12 as well as treatment options for this complication. RESULTS: We report the first non-European case of localized sclerodermoid reaction secondary to vitamin B12 injections. Our patient presented with disfiguring depression of the left upper arm following vitamin B12 injections for macrocytic anemia secondary to Crohn's disease. Histopathology showed findings similar to those seen in localized scleroderma. The patient was successfully treated with excision of diseased skin, local fat transfer, and complex repair with dramatic improvement at 5 months in contour and overall appearance. PMID- 15355373 TI - Local hatchet flap for facial skin defects reconstruction in special areas. AB - BACKGROUND: The local hatchet flap has been used to reconstruct skin defects of the face with satisfactory results. Cosmetically, it has advantages over other local flaps when skin defects are located in special areas such as the eyelid. OBJECTIVE: The purpose of this study was to examine the application of local hatchet flaps for facial defect reconstruction, especially in cases where reconstruction is classically difficult for cosmetic and functional rehabilitation. METHODS: We report an illustrated case series. Results. In this study, all four cases achieved satisfactory functional and cosmetic results using the local hatchet flap. CONCLUSIONS: The local hatchet flap is an effective and simple alternative for reconstructing facial defects in special areas of the face, offering good cosmetic results and satisfactory function. PMID- 15355374 TI - Multiple pigmented follicular cysts of the vulva successfully treated with CO2 laser: case report and literature review. AB - BACKGROUND: A pigmented follicular cyst is an uncommon epithelial cyst, typically occurring as a single asymptomatic pigmented papule or nodule on the head and neck of adult men. To the authors' knowledge, no previous cases of vulvar pigmented follicular cyst have been reported. Although benign, pigmented follicular cyst can affect quality of life when multiple and symptomatic. OBJECTIVE: The objective was to describe the clinicopathologic features of vulvar pigmented follicular cyst and to evaluate the efficacy of CO(2) laser therapy in this region. METHODS: A 62-year-old woman with multiple pigmented follicular cysts affecting the labia majora bilaterally is described. Owing to frequent inflammation and tenderness, continuous-wave CO(2) laser was used to treat the skin lesions under local anesthesia. RESULTS: All cystic nodules were successfully ablated with CO(2) laser. The patient tolerated the procedure well and had a satisfactory cosmetic outcome. CONCLUSION: CO(2) laser is a tolerable and effective therapeutic modality for multiple and symptomatic pigmented follicular cysts of the vulva. PMID- 15355375 TI - Nodular hidradenocarcinoma on the scalp of a young woman: case report and review of literature. AB - BACKGROUND: Nodular hidradenocarcinoma was first reported as clear-cell eccrine carcinoma by Keasby and Hadley in 1954 (Cancer 1954;7:934-52) and rare malignant tumor. Several synonyms and related terms for nodular hidradenocarcinoma have appeared in the literature. OBJECTIVE: They have potential for uncontrollable local recurrence, tend to metastasize, and often cause death. Most cases have been reported in the pathology literature with limited clinical information. METHODS: We report a 27-year-old woman with nodular hidradenocarcinoma on the scalp. RESULT: The management of rare cases is not well defined. In our case, she was only treated with a wide local excision, and no recurrence was observed 2 years after excision. CONCLUSION: Most authors have concluded that early wide surgical excision of the tumor is the treatment of choice. The efficiency of adjuvant therapy generally has not established. PMID- 15355376 TI - Cryosurgery for skin cancer. PMID- 15355377 TI - Noninvasive lower lid blepharoplasty with reply from Dr Ruiz-Esparza. PMID- 15355378 TI - Polidocanol or chromated glycerin for sclerotherapy of telangiectatic leg veins? With reply from Dr Kern et al. PMID- 15355380 TI - Neck-to-cheek transposition flap for a large cheek defect. PMID- 15355381 TI - Differences in the etiology of mandibular fractures in Kuwait, Canada, and Finland. AB - We studied causes of mandibular fractures treated in oral and maxillofacial units in three countries in years 1990-2000 in Kuwait (n=596), 1995-2000 in Canada (n=228), and 1990-99 in Finland (n=268). Of the Finnish patients, 27% were women. Corresponding percentages in Kuwait and Canada were 13 and 17%, respectively. Traffic crashes were the cause of injury in 55% of the cases in Kuwait and 33% in Oulu, but only 7% in Toronto. In Kuwait, the victims were often young people, which is why more traffic education, more control of speed, and more control of the use of safety belts should be implemented. Assault was the cause in 54% in Toronto, 12% in Kuwait, and 37% in Oulu. Falling was the cause in 22% of the cases in Kuwait. Alcohol was implicated in 21% of cases in Canada and 15% in Finland. PMID- 15355382 TI - Risk factors related to traumatic dental injuries in Brazilian schoolchildren. AB - The aim of this pilot study was to analyse whether overjet, lip coverage and obesity represented risk factors associated with the occurrence of dental trauma in the permanent anterior teeth of schoolchildren in Recife, Brazil. It included a random sample of 116 boys and girls aged 12 years, attending both public and private schools. Data was collected through clinical examinations and interviews. Dental trauma was classified according to Andreasen's criteria (1994). Overjet was considered as risk factor when it presented values higher than 5 mm. Lip coverage was classified as adequate or inadequate, while obesity was considered according to National Center for Health Statistics (NCHS) procedures for the assessment of nutritional status. The prevalence of dental injuries was 23.3%. Boys experienced more injuries than girls, 30 and 16.1%, respectively (P >0.05). There was a statistically significant difference between traumatic dental injuries and overjet (P <0.05) and between traumatic dental injuries and lip coverage (P=0.000). No statistical significant differences were found when obesity and dental trauma were analysed (P <0.05). It was concluded that boys from lower social strata attending public schools, presenting an overjet size greater than 5 mm and an inadequate lip coverage, were more likely to have traumatic dental injuries in Recife, Brazil. Obesity was not a risk factor for dental trauma in this sample. PMID- 15355383 TI - Prevalence of crown fractures in 8-10 years old schoolchildren in Canoas, Brazil. AB - The objective of this work was to evaluate the prevalence of children with crown fractures in permanent anterior teeth in 206 schoolchildren (104 girls and 102 boys) between the ages of 8 and 10 years, enrolled in three public schools in the city of Canoas, Brazil. The prevalence found was 17% with no significant difference between boys and girls, as well as between the ages. The most affected tooth was the maxillary central incisor, and a majority of the children showed only one affected tooth (88.6%). The types of fracture most commonly found were oblique and horizontal, and the portions of dental structures most affected were 'enamel only' and 'enamel and dentin'. Only seven children (20%) sought out dental treatment. PMID- 15355384 TI - Relationship between temporomandibular joint dynamics and mouthguards: feasibility of a test method. AB - A test system was developed establishing the feasibility of collecting biomechanical data as they relate to the use of mouthguards. Previous experimental studies have examined the physical and mechanical properties of mouthguard materials. This information has been used as a guide for establishing material standards and specifications for the fabrication of mouthguards, but it lacks the key biomechanical parameters required for a thorough mouthguard evaluation. The current study was designed to assess whether the impact force, condylar deflection, and strain superior to the temporomandibular joint region could be measured. A drop test was conducted on a cadaveric specimen to simulate loading at the chin point. To measure the force of impact, an accelerometer was attached to an impactor of known mass. High-speed biplanar (1000 frames per second) radiographs were used to determine condylar displacement. Radio-opaque markers were inserted into the bone at predetermined locations. Total displacement of these markers was determined in reference to anatomical landmarks. Strain gauges were attached to the mandible and skull to monitor the effects of the condyle impacting the base of the skull. Based on the data collected, forces were calculated by determining the product of the time-based acceleration and known mass. A measurable change in force between the mouthguards and the control (no mouthguard) was demonstrated. The average condylar displacement was successfully measured and indicated as an increase in total deflection for impacts conducted with mouthguards. Quantifiable strain was measured in the region above the mandibular fossa with and without the insertion of a mouthguard at all impact conditions. However, it was determined that additional gauges would provide critical data. Key biomechanical parameters for chin-point impacts were determined in the current study. The technique demonstrated that both displacement within the mandibular fossa and loading of the condyles occur during the impact event. Although the current study established a technique that can be used to examine the relationship between mouthguards and jaw-joint injuries, the role, if any, mouthguards play in the reduction of injuries cannot be established until a thorough analysis is completed. PMID- 15355385 TI - Conservative endodontic treatment of teeth fractured in the middle or apical part of the root. AB - According to treatment type, root-fractured teeth with pulp necrosis or exposed pulps were divided into five groups, group 1: 17 teeth in which the root canal of the coronal fragment only was filled with gutta-percha (GP); group 2: seven teeth in which the root canals of the coronal and apical fragments were both filled with GP; group 3: 19 teeth in which the coronal fragment was filled with GP and the apical fragment was surgically removed; group 4: 68 teeth where the root canal of the coronal fragment was treated with calcium hydroxide and subsequently filled with GP; and group 5: five vital teeth with root and concomitant crown fractures in which the exposed pulps were treated by partial pulpotomy. The frequency of periodontal healing was 76% in group 1, zero in group 2, 68% in group 3 and 86% in group 4. Compared with groups 1 and 2 combined, healing in group 4 was significantly more frequent. In groups 1, 2 and 4, failures occurred significantly more often in teeth showing overfilling, i.e. protrusion of GP into the space between the fragments, compared with teeth without overfilling. All five teeth in group 5 showed healing. It was concluded that root canal filling with GP of the coronal fragment only, with or without surgical removal of the apical fragment, can be successful in selected cases. Treatment of the root canal with calcium hydroxide followed by GP filling appears to be the treatment of choice in root-fractured non-vital teeth. Partial pulpotomy of exposed pulps in five teeth showed results similar to root-unfractured teeth with pulp exposure treated with this technique. PMID- 15355386 TI - Diagnostic characteristics of pulpal blood flow levels associated with adverse outcomes of luxated permanent maxillary incisors. AB - Laser Doppler flowmetry (LDF) is a non-invasive method to assess pulpal blood flow (PBF). Dental injury has been associated with significant PBF reduction. The purpose of this study was (i) to describe PBF characteristics of teeth with specific clinical outcomes, and (ii) to demonstrate diagnostic characteristics for different threshold PBF values for detection of specific multiple adverse outcomes. In 80 trauma patients, a single maxillary incisor treated by repositioning and splinting, and the respective contralateral homologous control tooth were investigated by LDF to assess local PBF values. Perfusion units (PU) were taken in two sessions, on the day of splint removal (session I), and 12 weeks after splint removal (session II). The ability of session II-related PBF measurements at 2.9, 6.4 and 9.9 PU levels to identify adverse outcomes occurring 36 weeks after splint removal was investigated. Adverse outcomes were classified as type I (loss of sensitivity), type II (periapical radiolucency), type III (grey discolouration), type IV (loss of sensitivity and periapical radiolucency), and type V (loss of sensitivity, periapical radiolucency and grey discolouration of crown). Receiver-operator characteristic (ROC) curves were used to evaluate the sensitivity and specificity of PBF assessments. There was a significant increase in PBF values from session I to session II (P=0.0001) for teeth without an adverse outcome, while teeth affected by a type II-V outcome showed a significant decrease in PBF values (P <0.05). PBF measurements did not change over time for the contralateral incisors (P >0.05). A type IV and V outcome occurred in 21 and 24% of the instances, respectively. The PBF of 2.9 PU demonstrated a sensitivity of 70% and a specificity of 93% for type V outcomes. The best likelihood ratio was found for the PBF 2.9 PU level and incisors associated with a type V outcome. The data suggest that the LDF test to be a valuable diagnostic adjunct for luxated teeth showing signs of adverse outcomes including grey discolouration or a combination of other signs. However, it may also become necessary to apply clinical decision-making methods in order to correctly evaluate the value of information gathered. The clinical implication is that LDF may become useful in the prediction of adverse outcomes at a much earlier time period than may be accomplished by standard sensitivity tests. PMID- 15355387 TI - Development of clinical and radiographic signs associated with dark discolored primary incisors following traumatic injuries: a prospective controlled study. AB - The purpose was to evaluate late complications of asymptomatic traumatized primary incisors with dark coronal discoloration. The clinical and radiographic signs of 97 teeth of the study group were recorded along a follow-up period that ranged between 12 and 75 months (mean >36 months). Children's age at time of injury ranged between 18 and 72 months (mean 40). The control group consisted of 102 non-discolored maxillary primary central incisors in 51 children older than 54 months with no history of dental trauma. In 50 teeth (52%) the color faded or became yellowish and in 47 (48%) it remained dark. Clinical signs of infection, that were diagnosed 5-58 months after the injury, were associated significantly more with dark than yellowish hues (83 and 17%, respectively). Teeth that had changed their color to become yellow presented more PCO than teeth with black/gray/brown coronal discoloration (78 and 6%, respectively). Arrest of dentine apposition was found in 15 teeth, one had yellow coronal discoloration and the remaining 14 had a dark shade. Eleven teeth showed inflammatory root resorption all with dark discoloration. Two atypical types of root resorption were observed: a surface resorption restricted to the lateral aspects of the apical half of the root while the root length remained unchanged and in the other expansion of the follicle of the permanent successor was observed. Expansion of the dental follicle was observed in 72% of all teeth with no significant difference between the various types of coronal discoloration but only half of the cases were associated with resorption of the root of the primary incisor. The various pathologic findings observed in the study group were either absent or rarely seen in the control group. It can be concluded that more than 50% of the primary incisors that retain their dark coronal discoloration acquired after dental injuries remain clinically asymptomatic till the eruption of the permanent successor even if they present accelerated root resorption. Asymptomatic traumatized primary incisors that retain their dark coronal discoloration may develop a sinus tract and inflammatory root resorption years after the injury. There is still a dilemma: which treatment is better for dark discolored primary incisors: early endodontic treatment or follow-up with the risk of development of infection and root resorption that may require extraction? PMID- 15355388 TI - Multiple complicated crown-root fracture of a permanent incisor. AB - The treatment of a transverse complicated and a vertical uncomplicated crown-root fractures with a horizontal root fracture of a maxillary right central incisor is presented. Coronal fragments were extracted atraumatically and gingivectomy was performed with electrosurgery to expose the subgingival fragment. The root canal was treated with calcium hydroxide to perform the healing at the fracture site. After the root fracture healing, a post was cemented into the canal and the coronal fragment was attached with a composite resin. Examination 18 months after treatment revealed good aesthetics and normal function. PMID- 15355389 TI - Multidisciplinary approach for the rehabilitation of dentoalveolar trauma. AB - Satisfactory dental rehabilitation of dentoalveolar trauma requires intense effort and time. Usually multidisciplinary treatment planning and teamwork are necessary to deal with multitask problems associated with these cases. Dental implants have been successfully used for replacement of missing teeth, but in trauma cases insufficient alveolar bone hinders implantation. In this report we present the multidisciplinary approach for the treatment of a trauma case. Maxillary segmental alveolar osteotomy in conjunction with interpositional and onlay bone grafting was performed to prepare the site for placement of osseointegrated implants. Titanium microplate and screws were used to provide orthodontic anchorage for intrusion of the extruded mandibular incisors. The patient was rehabilitated by implant supported fixed partial denture 6 months after implant placement. PMID- 15355390 TI - Localized edentulous ridge augmentation with upside down osteotomy prior to implant placement. AB - Localized bone defects may be seen following the extraction of teeth with periapical lesion or advanced periodontal disease, or as a result of trauma. When these regions are considered for treatment with implants, localized ridge augmentation will be necessary. Autogenous bone grafts are used exclusively for ridge augmentation. This case report represents the treatment of a localized edentulous ridge with an upside down osteotomy technique at the symphysis region prior to implant placement. Systemically healthy 21-year-old female patient, who was missing a lower right incisor tooth, was scheduled for an implant treatment. However, the crestal width was only 1 mm. The augmentation was planned and the region was treated with an upside down osteotomy technique. Nine months after the augmentation procedure, the computed tomography (CT) examination of the area revealed that the width of the crest was 7 mm, and the height of the crest was in good relation with the cementoenamel junction of the adjacent teeth. Flipping a bone block graft, which was harvested from the edentulous area, upside down may provide a successful result in partially edentulous ridges, in both maxilla and mandible. PMID- 15355391 TI - Titanium release from implants prepared with different surface roughness. AB - OBJECTIVES: There may be a risk of greater ion release for surface-enlarged implants than conventionally turned components. The major aim of the present paper was to investigate whether a correlation exists between ion release and a surface roughness relevant for today's commercial implants. Other aims were to compare ion release after two insertion times and concentration in bone tissue as a function of distance from the implant surface. MATERIAL AND METHODS: Lactic acid aqueous solution (pH=2.3) and phosphate-buffered saline were used for the in vitro investigation. For the in vivo investigation, synchrotron radiation X-ray fluorescence (SRXRF) spectroscopy and secondary ion mass spectroscopy (SIMS) were performed 12 weeks and 1 year after implantation in rabbit tibiae. RESULTS: The average height deviation (S(a)) was 0.7, 1.27, 1.43 and 2.21 microm, respectively, for the four surfaces investigated. No difference in ion release was found in vitro. In vivo, SRXRF demonstrated slightly higher values for the roughest surface up to a distance of 400 microm from the implant surface; thereafter no difference was found. SIMS demonstrated no difference in ion release for the roughest and smoothest surfaces, but slightly more titanium in bone tissue after 1 year than after 12 weeks. Titanium rapidly decreased with distance from the implant surface. CONCLUSION: At a level relevant for commercial oral implants, no correlation was found between increasing roughness and ion release, neither in vitro nor in vivo. PMID- 15355392 TI - Morphologic, functional and behavioral effects of titanium dioxide exposure on nerves. AB - OBJECTIVES: The purpose of this study was to explore morphologic, functional, and behavioral effects of titanium dioxide (TiO(2)) on nerves. MATERIAL AND METHODS: A total of 17 albino rats were used for nerve conduction experiments, hot-plate tests, and histological evaluation. TiO(2) was implanted unilaterally on the sciatic nerves of five rats. Ten days after surgery, test and control nerves were dissected and their signal transduction speeds were quantified by suction electrodes in a bath containing a Tyrode solution. Twelve rats were divided into three equal groups resulting in equal number of nerves (n=8) for TiO(2) implantation, surgical exposure of the nerves, and for use as controls. One week after surgery, hot-plate tests were undertaken for 10 consecutive days to determine response latencies of the nerves. At the termination of the experiments, the nerves were harvested, processed, and examined under a microscope. RESULTS: The signal transduction speeds of TiO(2)-implanted nerves was similar to control specimens (P>0.05). The avoidance responses of TiO(2) implanted, surgically exposed, and control nerves were comparable (P>0.05). At the cellular level, TiO(2) did not lead to any signs of adverse reactions on nerves. CONCLUSIONS: TiO(2), the main oxide surrounding endosseous titanium implants, does not alter the structure and the function of myelinated nerves. PMID- 15355393 TI - Predicting osseointegration by means of implant primary stability. AB - The purpose of the present clinical study was (1) to evaluate the Osstell as a diagnostic tool capable of differentiating between stable and mobile ITI implants, (2) to evaluate a cut-off threshold implant stability quotient (ISQ) value obtained at implant placement (ISQitv) that might be predictive of osseointegration, (3) to compare the predictive ISQitv of immediately loaded (IL) implants and implants loaded after 3 months (DL). Two patient groups were enrolled, 18 patients received 63 IL implants and 18 patients were treated with 43 DL implants. The ISQ was recorded at implant placement, after 1, 2, 4, 6, 8, 10 and 12 weeks. All implants passed the 1-year loading control. Two implants failed, one DL implant with ISQ at placement (ISQi) of 48 and one IL implant with ISQi of 53. The resonance-frequency analysis (RFA) method was not a reliable diagnostic tool to identify mobile implants. However, implant stability could be reliably determined for implants displaying an ISQ> or =47. After 1 year of loading, all DL implants with an ISQi> or =49 and all IL implants with an ISQi> or =54 achieved and maintained osseointegration. By the end of 3 months, implants with ISQi<60 had an increase of stability. Implants with ISQi 60-69 had their stability decrease during 8 weeks before returning to their initial values. Implants with ISQ>69 had their stability decrease during the first 4 weeks before remaining stable. Although preliminary, these data might orient the practitioner to choose among various loading protocols and to selectively monitor implants during the healing phase. PMID- 15355394 TI - Implant stability measurement of delayed and immediately loaded implants during healing. AB - The purpose of the present study was (1) to measure the primary stability of ITI implants placed in both jaws and determine the factors that affect the implant stability quotient (ISQ) determined by the resonance frequency method and (2) to monitor implant stability during the first 3 months of healing and evaluate any difference between immediately loaded (IL) implants and standard delayed loaded (DL) implants. The IL and DL groups consisted of 18 patients/63 implants and 18 patients/43 implants. IL implants were loaded after 2 days; DL implants were left to heal according to the one-stage procedure. The ISQ was recorded with an Osstell apparatus (Integration Diagnostics AB, Gothenburg, Sweden) at implant placement, after 1, 2, 4, 6, 8, 10 and 12 weeks. Primary stability was affected by the jaw and the bone type. The ISQ was higher in the mandible (59.8+/-6.7) than the maxilla (55.0+/-6.8). The ISQ was significantly higher in type I bone (62.8+/-7.2) than in type III bone (56.0+/-7.8). The implant position, implant length, implant diameter and implant deepening (esthetic plus implants) did not affect primary stability. After 3 months, the gain in stability was higher in the mandible than in the maxilla. The influence of bone type was leveled off and bone quality did not affect implant stability. The resonance-frequency analysis method did not reveal any difference in implant stability between the IL and DL implants over the healing period. Implant stability remained constant or increased slightly during the first 4-6 weeks and then increased more markedly. One DL and IL implant failed; both were 8 mm long placed in type III bone. At the 1-year control, the survival rate of the IL and the DL implants was 98.4% and 97.7%, respectively. This study showed no difference in implant stability between the IL and DL procedures over the first 3 months. IL short-span bridges placed in the posterior region and full arch rehabilitation of the maxilla with ITI sandblasted and-etched implants were highly predictable. PMID- 15355395 TI - Histological evidence of osseointegration in human retrieved fractured hydroxyapatite-coated screw-type implants: a case report. AB - Histological evidence of clinically successful dental implants is very rare. This case report presents histological evidence of osseointegration in human implants retrieved because of fractures at the connected portion between the abutments and fixtures due to a car accident. The duration of functional loading of the implants was 18 months. Two hydroxyapatite (HA)-coated screw-type implants were removed with part of the healthy bone from the mandibular left molar region. A block was prepared using cutting and grinding equipment to obtain a central section approximately 50 microm thick, which was stained with basic fuchsin and methylene blue. Histological examination revealed that the bone was dense and in close relation with the HA coating of the implants. The interspaces of each thread of the implant were filled with mineralized bone. Peri-implant soft tissue was not observed in the section. A high degree of osseointegration was noted, with a bone-to-implant contact of 87.5% (implant corresponding to tooth 36) and 97.4% (implant corresponding to tooth 37). The connection between the 30 and 50 microm HA coating and the metal was uniformly tight and constant. In conclusion, the histological evidence showed a high degree of osseointegration in two HA coated screw-type dental implants retrieved after functional loading for 18 months. PMID- 15355396 TI - Correlation of peri-implant health and myeloperoxidase levels: a cross-sectional clinical study. AB - OBJECTIVES: At present, there are no diagnostic tools that permit early detection of peri-implantitis. The purpose of this cross-sectional study was to evaluate the correlation of myeloperoxidase (MPO) levels with traditional periodontal clinical parameters around dental implants including peri-implant pocket probing depth (PPD), gingival index (GI) and bleeding on probing (BOP), since MPO has been associated with destruction of periodontal tissues. MATERIAL AND METHODS: Twenty-four healthy adult volunteers (9 men and 15 women) with 64 Ankylos Biofunctional implants (DentsplyFriadent, Mannheim, Germany) were recruited from Tallinn Dental Clinic. Biochemical and clinical parameters evaluated were the following ones: the level of MPO in the peri-implant sulcus fluid (PISF) (an analog for gingival crevicular fluid in natural teeth), PPD (mm), GI (0,1,2 or 3), and BOP (0 or 1). RESULTS AND CONCLUSION: In comparison to the clinically healthy implants, total amounts of MPO were significantly higher in PISF collected around implants with inflammatory lesions. In addition, the levels of MPO were correlated with the clinical parameters. The results confirm the similarity of the inflammatory response of tissues surrounding implants and natural teeth, and suggest that MPO could be promising marker of inflammation around dental implants. PMID- 15355397 TI - Immunohistochemical analysis of soft tissues in implants with healthy and peri implantitis condition, and aggressive periodontitis. AB - Today, implant-supported prostheses are widely accepted as a reliable treatment modality, but failures in longitudinal studies have been shown. In some cases, peri-implantitis with a progressive periodontal bone loss takes place, and mechanical or load factors and biological or plaque-induced lesions have been claimed as main etiologic factors. We compared five cases of peri-implantitis, with five cases of healthy peri-implant tissues and five cases of aggressive periodontitis in order to give new findings on the osseointegration loss process. Biopsy specimens from the peri-implant tissues including oral (O), sulcular, and junctional epithelium and the underlying and supracrestal connective tissue, were taken in all cases for histological and immunohistochemical analysis. T lymphocytes were the most prominent cell in the peri-implantitis (PG) and aggressive periodontitis (AG) groups, but not in the peri-implant healthy group (HG). CD1a-positive cells (Langerhans and immature dendritic cells) were observed more frequently in the O than in the sulcular-junctional (S-J) epithelium: they were located in the basal and parabasal layers, without any differences between the three groups. Vascular proliferation analysed by immunoreactivity for CD34, Factor VIII, and vascular endothelial growth factor was more prominent in the PG comparing with HG and AG in the S-J area. Apoptosis, analysed by bcl2 and p53 immunoreactivity, was similar in the three groups. In conclusion, we suggest that the osseointegration loss process is due to an inflammatory process similar to that observed in aggressive periodontitis according to the number of T lymphocytes, but not to the vascular proliferation. PMID- 15355398 TI - Mandibular two-implant telescopic overdentures. AB - To stabilize mandibular overdentures in edentulous patients, various connector types which can be attached to between two and four implants placed in the anterior mandible are possible. Treatment using non-rigid telescopic connectors on two interforaminal implants for overdenture stabilization began in 1989. The objective of this study is to investigate soft- and hard-tissue conditions as well as prosthesis function after a period of 10 years. This also involved an evaluation of correlations between radiographic and clinical parameters. Twenty three subjects with 46 interforaminal implants (ITI solid screw implants, 12 mm in length, 4.1 mm in diameter; 10.4 years in situ, range, 8-12.8 years) were investigated. Modified plaque index (mPI), sulcus fluid flow rate (SFFR), modified sulcus bleeding index (mBI), probing depth (PD), distance from implant crown margin to the coronal border of the peri-implant mucosa (DIM), attachment level (AL), width of keratinized mucosa (KM), Periotest values (PTVs) and prosthesis function were evaluated. In the radiographic evaluation, the distance between implant shoulder and first crestal bone-implant contact (DIB) in mm and the horizontal bone loss (HBL) in mm were measured. The relatively high mPI scores (mean, 0.82; score, 0 in 44.4%; SD, 0.83) did not result in increased SFFR scores (mean, 12; min, 3, max, 38; SD, 7.43) or higher mBI scores (mean, 0.35; score, 0 in 70.8%; SD, 0.59), which was commensurate with healthy peri-implant mucosa. A mean PD value of 2.15 mm (min, 1 mm; max, 5 mm; SD, 0.96) and a mean DIM value of 0.28 mm (min, 0 mm; max, 2 mm; SD, 0.52) were measured. The implants were stable, showing a mean Periotest value of -1.91 (max, 02, min, -6; SD, 1.76). A mean DIB of 3.19+/-0.95 mm (range, 1.3-5.16 mm) and a mean HBL of 1.6+/ 1.52 mm (range, 0.28-8.33 mm) were calculated. A correlation was found between DIB and the parameters SFFR (P=0.060), DIM (P=0.042), AL (P=0.050) and especially PTV (P<0.01), leading to the assumption that these clinical parameters may be useful indicators of peri-implant bone loss. The results of the 10-year follow-up examination show that non-rigid telescopic connectors with two interforaminal implants for overdenture stabilization appear to be an efficient and effective long-term treatment modality in severely resorbed edentulous mandibles. Particularly in geriatric patient treatment this concept may provide advantages in terms of handling, cleaning and long-term satisfaction. PMID- 15355399 TI - Measurement accuracy of reconstructed 2-D images obtained by multi-slice helical computed tomography. AB - The present dental reconstructed computed tomography (CT) images consist of continuously piled-up axial CT images that are perpendicular to the axial CT plane. In the mandibular posterior region, the angles of designed implants frequently differ from the angles perpendicular to the axial CT plane and the measurement of mandibular height is inaccurate. This study was performed to clarify the measurement accuracy of double-oblique reconstructed images in multi slice helical CT, using dried mandibles. The mandibular height was measured from the alveolar crest immediately below the aluminum tube to the superior border of the mandibular canal using the double-oblique reconstructed images and the micro CT average images of three semi-lateral dried mandibles. The mean of the differences between the double-oblique reconstructed images with a table pitch of 1.5 in a helical scan and the micro-CT average images was 0.31 mm. It is concluded that the accuracy of the double-oblique reconstructed image measurements with helical scan is high. So this double-oblique program can be applied to the imaging diagnosis in dental implant treatment. PMID- 15355400 TI - Porcine sinus mucosa holds cells that respond to bone morphogenetic protein (BMP) 6 and BMP-7 with increased osteogenic differentiation in vitro. AB - The aim of this in vitro study was to determine whether the sinus mucosa holds cells with an osteogenic potential. Frozen sections of sinus mucosa from three adult pigs were investigated for the expression of STRO-1, a marker of mesenchymal progenitor cells, and alkaline phosphatase activity, an enzyme expressed by cells committed to the osteogenic lineage and by mature osteoblasts. To determine their osteogenic potential, mucosa-derived cells were incubated with bone morphogenetic protein (BMP)-6 and BMP-7, and alkaline phosphatase activity, osteocalcin expression, and mineralization of the extracellular matrix was measured. We found sinus mucosa cells staining positive for STRO-1 and alkaline phosphatase activity. When sinus mucosa tissue was placed in culture, alkaline phosphatase positive cells grew out from the explants and further increased alkaline phosphatase activity in response to BMP-6 and BMP-7. The expression level of the osteoblast-specific extracellular matrix protein osteocalcin, and the amount of calcium accumulation within the extracellular matrix was also increased in response to BMPs. We conclude that the sinus mucosa holds mesenchymal progenitor cells and cells committed to the osteogenic lineage that can respond to BMP-6 and BMP-7 by an increase of their osteogenic differentiation. PMID- 15355401 TI - Effects of platelet lysates on select bone cell functions. AB - Although platelet-rich plasma and platelet concentrates have been used to promote bone healing in orthopaedic and maxillofacial surgery, the underlying cellular level mechanisms remain poorly understood. The present in vitro study investigated the effects of human platelet lysate (PL) on selected functions of cultured bone cells. Cells from 18-day-old fetal rat calvaria were isolated by a collagenase digestion procedure. PL was added at different concentrations on pre- or post-confluent cell stage. After 1 day, bone cell proliferation was maximal and half-maximal in the presence of PL from 3 x 10(8) and 0.5 x 10(8) platelets/ml, respectively. During 17 h, the number of bone cells traversing the scrape border of a scrape wound model increased by 16-fold in the presence of PL from 3 x 10(8) platelets/ml. The presence of PL from 3 x 10(8) platelets/ml in pre-confluent bone cell cultures for 48 h resulted in a threefold decrease of alkaline phosphatase (ALP) specific activity. In the case of confluent bone cells, the presence of PL (from 1 x 10(6) to 3 x 10(8) platelets/ml) for 11 days, the ALP specific activity and total calcium content decreased in a PL dose dependent manner and reached a minimum in the presence of PL from 3 x 10(8) platelets/ml. In summary, short-term PL exposure (up to 24 h) promotes the proliferative and chemotactic bone cell functions while long-term PL exposure results in a decrease of both ALP activity and mineral formation. These data show that the soluble components contained in PL may affect the bone healing process by modulating differently bone cell functions. PMID- 15355402 TI - Tissue-engineered injectable bone regeneration for osseointegrated dental implants. AB - The present study investigated a correlation between osseointegration in dental implants and an injectable tissue-engineered bone, using mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP). Initially, the teeth in the mandible region were extracted and the healing period was 1 month. Bone defects on both sides of the mandible were prepared with a trephine bar. The defects were implanted with graft materials as follows: PRP, dog MSCs (dMSCs), and PRP, autogenous particulate cancellous bone and marrow (PCBM), and control (defect only). Two months later, the animals were evaluated by histology, and at the same time dental implants were installed. Two months later, the animals were sacrificed and nondecalcified sections were evaluated histologically and histometrically. According to the histological observations, the dMSCs/PRP group had well-formed mature bone and neovascularization, compared with the control (defect only) and PRP groups, as was the same for the PCBM group. A higher marginal bone level was observed around implants with PRP, PCBM, and dMSCs/PRP compared with the control. Furthermore, the values describing the amount of bone implant contact (BIC) at the bone/implant interface were significantly different between the PRP, PCBM, dMSCs/PRP, and control groups. Significant differences were also found between the dMSCs/PRP and control groups in bone density. The findings of this experimental study indicate that the use of a mixture of dMSCs/PRP results in good results such as the amount of BIC and bone density comparable with that achieved by PCBM. PMID- 15355403 TI - Reconstruction of maxillary and mandibular defects using prefabricated microvascular fibular grafts and osseointegrated dental implants -- a prospective study. AB - The fibular flap can be used for a variety of indications. Recently, the treatment of four patients with severely atrophied upper jaws using a method to prefabricate the vascularized fibular graft has been published. This technique consists of a two-stage operation procedure that allows simultaneous prosthodontic rehabilitation and immediate placement of dental implants. In this paper eight patients with 29 ITI implants (Straumann AG, Waldenburg, Switzerland) who had reconstruction of either the upper or lower jaw are presented. The aim of the study was (i) to evaluate the behavior of the newly formed soft tissue around implants inserted in the fibula by applying periodontal parameters, (ii) to monitor prospectively the integration of the implants in the fibular graft, and (iii) to assess the osseous integration of the fibular graft used for reconstruction of the upper or lower jaw. Two implants failed during the observation time because of avascular bone at the distal end of the fibular graft. Stabilization of the graft, however, was never compromised. Due to the prefabrication firmly attached gingiva-like soft tissue could be provided preventing periimplant soft tissue inflammation and facilitating oral hygiene. After 1 year of observation the mean attachment level was similar to implants placed in original bone whereas vertical bone loss measured radiographically was lower in the present study. This may indicate that the remodeling of a bicortical bone requires a longer period of time compared with the bone of the alveolar crest. The prospective 1-year results are promising but long-term evaluation of periodontal and radiological parameters are required. PMID- 15355404 TI - Extensive augmentation of the alveolar ridge using autogenous calvarial split bone grafts for dental rehabilitation. AB - Free autogenous iliac bone is the most commonly used graft material for an extensive alveolar ridge reconstruction. The application of iliac bone, however, is associated with problems, such as transplant loss resulting from postoperative infection and late bone resorption. A bone-graft material more suitable than iliac bone is therefore still needed. This paper describes a concept for alveolar ridge reconstruction using calvarial split bone, and the related surgical techniques. Clinical and radiological follow-up examinations were undertaken to evaluate the potential benefit of calvarial split bone in alveolar-ridge reconstruction. Between 1999 and 2002, 13 patients with a mean age of 54 years (range 31-70 years) underwent surgery, seven patients in the maxilla and six in the mandible. In four cases, wound dehiscence occurred postoperatively. In one of these cases, the dehiscence was associated with a local infection. However, no bone transplants were lost. After a mean follow-up time of 19.6 months, bone resorption, measured radiologically, was minimal. Endosseous dental implants were successfully installed and maintained. Satisfactory prosthetic rehabilitation was achieved in all patients. Our preliminary experience suggests that calvarial split bone may be regarded as a promising alternative to autogenous iliac bone in connection with extensive augmentation of the alveolar ridge. PMID- 15355405 TI - Anterior tooth replacement with implants in grafted alveolar cleft sites: a case series. AB - When a residual edentulous space in the anterior region remains after surgical and orthodontic treatment of alveolar cleft patients, implant placement with or without additional grafting is a treatment option. We placed a total of 10 implants in nine consecutive alveolar cleft patients (five females, four males) with residual anterior edentulous spaces and non-restored neighbouring teeth. Patient age generally varied from 18 to 22 years. Additional (tertiary) bone grafting, implant placement after 3 months of graft consolidation and implant uncover after no less than 6 months was the favoured surgical protocol in the five most recent cases. Ample time was allowed to evaluate and guide the development of soft tissues, following second-stage surgery. For this purpose, temporary crowns were fabricated, adjusted and left in place for over 3-4 months in seven out of nine cases. No implants were lost after a mean observation period of 3.4 years and all implants function without objective (radiographic) or subjective problems. Aesthetics were considered acceptable in seven out of nine cases. Unsatisfactory aesthetic results originated from poor implant positioning or alignment in two patients who did not undergo tertiary bone grafting. We conclude that implant placement in alveolar cleft sites is a comprehensive but viable treatment option. Tertiary grafting of the site is recommended to achieve better ridge contour and bone height. This allows optimal implant placement and alignment. PMID- 15355406 TI - Involvement of Fusarium spp. in fungal keratitis. AB - Members of the filamentous fungal genus Fusarium are among the agents most frequently causing keratomycosis in humans. Fusarium keratitis is most common among agricultural workers in geographical regions with hot, humid, tropical or semi-tropical climates, but can occur more rarely in countries with temperate climates, such as Hungary. Keratitis is usually treated with a topical antifungal agent, sometimes in combination with sub-conjunctival injections and/or antimycotic agents, but therapeutic keratoplasty may be needed for patients whose corneal infection does not resolve. Early and accurate diagnosis, coupled with appropriate antifungal therapy, is crucial for improving the chances of complete recovery. PMID- 15355407 TI - Beta-lactamase-producing bacteria in mixed infections. AB - Beta-lactamase-producing bacteria (BLPB) can play an important role in polymicrobial infections. They can have a direct pathogenic impact in causing infections, as well as an indirect effect through their ability to produce the beta-lactamase. BLPB may not only survive penicillin therapy themselves, but can also protect other penicillin-susceptible bacteria from penicillin by releasing free beta-lactamase into their immediate environment. This phenomenon occurs in upper respiratory tract, skin, soft tissue, surgical and other infections. The in vitro and in-vivo clinical evidence supporting the role of BLPB in the increasing failure of penicillin to resolve such infections, and the implications of this phenomenon for the management of infections, are discussed. PMID- 15355408 TI - Assessment of nephrotoxicity in patients receiving amphotericin B lipid complex: a pharmacosurveillance study in Spain. AB - This study assessed the risk of haematological, renal and hepatic toxicity associated with amphotericin B lipid complex (ABLC; Abelcet) in a multicentre, open-label, non-comparative study of 93 patients from 17 different hospitals who received ABLC because of proven or suspected systemic fungal infection or leishmaniasis. Most (66%) patients had onco-haematological diseases. Optimum treatment with ABLC comprised a slow (2-h) infusion dose of 5 mg/kg/day for a minimum period of 14 days. Biochemical and haematological parameters were measured pre-, during and post-treatment. In the overall patient group, the mean serum creatinine concentration was similar pre- and post-study (1.00 +/- 1.14 mg/dL vs. 1.20 +/- 1.19 mg/dL; p > 0.05). There were no significant changes pre- and post-treatment in concentrations of haemoglobin, potassium, transaminases and bilirubin. There was no significant correlation between the dose administered and the concentrations of serum creatinine (Spearmann 0.22). There was no greater nephrotoxicity in the patients with previous renal failure, or in those who had received amphotericin B previously. There were serious adverse events in five patients, but other alternative causes that could explain these events were present in three of these patients. Fevers or chills were experienced by 23% of the patients during the ABLC infusion, but only in one case did this necessitate the suspension of treatment. It was concluded that ABLC is a drug with low nephrotoxicity, even when administered to patients with pre-existing renal insufficiency. Adverse events were generally slight or moderate, and were managed easily with appropriate pre-medication. PMID- 15355409 TI - Serotypes and clinical manifestations of invasive group B streptococcal infections in western Sweden 1998-2001. AB - This study monitored the serotypes of Streptococcus agalactiae (group B streptococcus; GBS) isolated from invasive infections in western Sweden and investigated possible relationships between serotype, age and clinical manifestations. Invasive GBS isolates were collected prospectively during 1998 2001 at six laboratories, covering two counties with a population of 1.8 million, and were serotyped by coagglutination. Clinical data were obtained from hospital notes. In total, 161 invasive strains (50 from neonates and infants aged < 3 months, and 111 from adults) were serotyped. The commonest serotypes from neonates and infants were serotypes III (60%), V (22%) and Ia (10%), and from adults were serotypes V (42%) and III (25%). Serotype V had doubled in frequency among both children and adults compared to a previous study from the same area in 1988-1997. Most (80%) of the adults had an underlying medical condition. No relationship was found between serotype and clinical manifestations. However, the study demonstrated the importance of active surveillance of GBS serotypes and the difficulties of formulating a multivalent polysaccharide conjugate vaccine against GBS. PMID- 15355410 TI - A cross-sectional survey of the prevalence of Streptococcus pneumoniae nasopharyngeal carriage in Belgian infants attending day care centres. AB - Nasopharyngeal carriage is a major factor in the transmission of pneumococcal disease. The aim of this study was to determine the prevalence of asymptomatic nasopharyngeal carriage of Streptococcus pneumoniae and the distribution of serogroups and serotypes in children aged 3-36 months attending day care centres in Belgium. A single nasopharyngeal swab was cultured from 467 children attending 30 different day care centres between December 2000 and March 2001. S. pneumoniae isolates were serotyped and their antibiotic susceptibilities assessed by disk diffusion. The overall nasopharyngeal carriage rate for S. pneumoniae was 21% in the 467 children. None of the commonly accepted risk factors studied was associated significantly with carriage. Capsular serotypes isolated were 19F (27.3%), 6B (20.2%), 23F (19.2%), 19A (10.1%), 6A (7.1%), 14 (5.1%) and others (11.0%). Theoretical coverage by the seven-valent (serotypes 4, 6B, 9V, 14, 18C, 19F and 23F) pneumococcal conjugate vaccine was 73.7%. Fourteen (14.1%) of 99 strains were non-susceptible to penicillin, 48 (48.5%) to tetracycline and 61 (61.6%) to erythromycin. Theoretical coverage by the seven-valent pneumococcal conjugate vaccine was 93% for the penicillin-resistant serotypes, 69% for the tetracycline-resistant serotypes and 75% for the erythromycin-resistant serotypes. Use of the seven-valent pneumococcal conjugate vaccine could potentially reduce nasopharyngeal carriage of the antibiotic-resistant strains. PMID- 15355411 TI - Antimicrobial susceptibility trends among Escherichia coli and Shigella spp. isolated from rural Egyptian paediatric populations with diarrhoea between 1995 and 2000. AB - Antimicrobial susceptibility testing was performed on 3,627 isolates of Escherichia coli and 180 isolates of Shigella spp. collected in rural locations from 875 Egyptian children with diarrhoea between 1995 and 2000. The cumulative rates of resistance for E. coli and Shigella spp. were high (respectively, 68.2% and 54.8% for ampicillin, 24.2% and 23.5% for ampicillin-sulbactam, 57.2% and 42.5% for trimethoprim-sulphamethoxazole, and 50.9% and 75.4% for tetracycline). Non-enterotoxigenic E. coli (NETEC) isolates had a consistently higher level of antimicrobial resistance than did enterotoxigenic E. coli (ETEC) isolates. Trend testing showed significant decreases in resistance to ampicillin, ampicillin sulbactam and tetracycline among all E. coli isolates. Increasing rates of resistance were observed for trimethoprim-sulphamethoxazole in ETEC isolates and Shigella spp., but not in NETEC isolates. Low levels of resistance were observed for all other antimicrobial agents tested. Overall, high levels, but decreasing trends, of resistance to commonly used antimicrobial agents were detected among isolates of E. coli and Shigella spp. from children in rural Egypt. PMID- 15355412 TI - Genetic characterisation of Helicobacter pylori isolates from an Argentinean adult population based on cag pathogenicity island right-end motifs, lspA-glmM polymorphism and iceA and vacA genotypes. AB - Isolates of Helicobacter pylori from 88 patients were characterised by cagA status, cagA pathogenicity island (PAI) right-end motifs, iceA, vacA and lspA glmM genotypes, primarily by PCR-based analysis, to investigate whether Argentinean isolates differed from those recovered in southern Europe or other Latin American countries. PCR-based analysis of vacA alleles was confirmed by reverse hybridisation in 56 cases, while sequence analysis was performed either when iceA and vacA genotypes could not be determined by PCR, or to investigate PCR and reverse hybridisation vacA genotyping discordance. Typing by lspA-glmM restriction fragment length polymorphism was performed with HhaI and AluI. The pattern of cag PAI right-end motifs and the prevalence of type Ia were similar to those in isolates from southern European countries, with cagA(+)/iceA1/vacA-s1 m1 being the commonest genotype. Reverse hybridisation identified a vacA-s1a/s1b recombinant allele, confirmed by sequencing analysis. Analysis of lspA-glmM genotypes identified at least 73 unrelated strains. Few mixed infections were identified, but in one case, isolates from a single biopsy exhibiting two vacA alleles were shown by lspA-glmM fingerprints to be two unrelated strains. No associated effect on ulcer disease risk was demonstrated by analysis of cagA, vacA and iceA status. Overall, the isolates of H. pylori from Argentina were similar to isolates from southern Europe or Latin American countries, and infections were associated mainly with single H. pylori strains. PMID- 15355413 TI - Chlamydia pneumoniae in HIV-infected patients and controls assessed by a novel whole blood interferon-gamma assay, serology and PCR. AB - Chlamydia pneumoniae seropositivity is associated with cardiovascular disease and HIV infection. Cell-mediated immune responses are important for control of C. pneumoniae, and such responses may be impaired in HIV-infected patients. An assay for detection of interferon (IFN)-gamma in whole blood stimulated with C. pneumoniae antigen was developed and studied in HIV-infected patients and uninfected controls. Among 34 HIV-infected patients, none had an IFN-gamma response to C. pneumoniae antigen, compared with five of 32 healthy controls (p < 0.001). Fewer HIV-infected individuals elicited a serum IgG response when tested with a commercial enzyme immunoassay (p 0.009), but this was not so for serum IgA (p 0.12). Additionally, the IFN-gamma and antibody assays showed a trend towards a bivariate response in normal controls. This indicates that cellular and antibody responses against C. pneumoniae may be mutually exclusive, with potential implications for the role of this organism in the genesis of cardiovascular disease in both immunocompetent and HIV-infected populations. PMID- 15355414 TI - Prevalence of drug resistance and newly recognised treatment-related substitutions in the HIV-1 reverse transcriptase and protease genes from HIV positive patients naive for anti-retrovirals. AB - The aim of this study was to assess the prevalence of genetic changes in either the HIV reverse transcriptase (RT) or protease (Pro) genes in a cohort of patients naive for anti-retroviral therapy. Of 61 patients, 43 (70.5%) were infected with HIV strains harbouring at least one resistance-related mutation, with 41 (67.2%) harbouring newly recognised treatment-related mutations. Among the 61 patients, the prevalence of specific mutations in the RT gene was as follows: 39A, 1.6%; 43E, 1.6%; and 228H, 1.6%. The prevalence of specific mutations in the Pro gene was as follows: 11I, 1.6%; 13V, 26.2%; 35D, 19.6%; 45R, 1.6%; 58E, 1.6%; 62V, 31%; 72V, 11.4%; 72M, 6.5%; 72T, 3.2%; 75I, 1.6%; and 89M, 13%. A higher prevalence of newly recognised mutations was found in strains from patients infected through sexual practices (30/36 = 83.4% vs. 11/25 = 44%; p 0.0023; OR 10.91; 95% CI 3.14-40.39). These findings support the use of resistance testing in patients naive for anti-retroviral therapy, and suggest that the possible impact of newly recognised treatment-related mutations on clinical outcome requires further investigation. PMID- 15355415 TI - Conservative medical therapy of prosthetic joint infections: retrospective analysis of an 8-year experience. AB - Successful treatment of prosthetic joint infections often requires multiple surgical interventions and prolonged antimicrobial therapy. However, in certain situations, a surgical approach may not be in the best interest of the patient. A conservative approach was used to treat 34 patients with prosthetic joint infection between 1995 and 2003. Diagnosis of infection was based on clinical microbiological evidence, confirmed by (99)Tc-labelled leukocyte scintigraphy, and involved 12 Staphylococcus aureus infections, nine Staphylococcus epidermidis infections, two Enterococcus faecalis infections, two mixed infections (S. aureus plus Pseudomonas aeruginosa; S. epidermidis plus E. faecalis), with the infecting pathogen being unidentified for nine patients. Most infections were treated initially with intravenous or intramuscular teicoplanin +/- ciprofloxacin or rifampicin, followed by oral ciprofloxacin or minocycline plus rifampicin. The mean duration of antimicrobial therapy was 41.2 weeks. Overall, only three patients did not respond to therapy, and infection was controlled in the remaining 31 patients. Among these, no relapse was observed in 17 patients during follow-up for 9-57 months; improvement with early (within 6 months of antibiotic discontinuation) or late relapse was observed in seven and three patients, respectively; two patients improved clinically, but continued to receive antibiotic therapy; and two patients whose condition improved initially were lost after a 6-month follow-up following discontinuation of antibiotics. No patient complained of side effects requiring discontinuation of antibiotic therapy. The study confirmed that suppression of infection, with salvage of the infected device in an acceptably functional state, can be achieved in selected cases. PMID- 15355416 TI - A European perspective on intravascular catheter-related infections: report on the microbiology workload, aetiology and antimicrobial susceptibility (ESGNI-005 Study). AB - The laboratory workload, microbiological techniques and aetiology of catheter related infections in European hospitals are mostly unknown. The present study (ESGNI-005) comprised a 1-day (22 October 2001), laboratory-based, point prevalence survey based on a questionnaire completed by microbiology laboratories in European (European Union (EU) and non-EU) hospitals. Also included were questions requesting retrospective information for the year 2000. In total, 151 hospitals from 26 European countries participated, of which 78.1% were teaching institutions. Overall, the estimated population served by these institutions was 121,363,800, and the estimated number of admissions during 2000 was 6,712,050. The total number of catheter tips processed during 2000 was 142,727, or 21/1,000 admissions, of which 23.7% were considered to be positive in the institutions using semiquantitative or quantitative techniques. Overall, EU centres received significantly more catheter tip samples/1,000 admissions and had a significantly higher rate of 'positivity' (p < 0.0001) than non-EU centres. Of the institutions surveyed, 11.4% (7.2% in EU countries and 23.7% in non-EU countries; p 0.04) used only qualitative techniques for catheter tip sample processing. On the day of the study, 167 microorganisms were recovered from significant catheter tip cultures (122 patients), of which Gram-positive bacteria represented 70.7%, Gram-negative bacteria 22.2%, and yeasts 7.2%. The five most common microorganisms were coagulase-negative staphylococci, Staphylococcus aureus, Candida spp., Enterococcus spp. and Pseudomonas spp. Overall, 19% of catheter tip cultures were polymicrobial. In the case of S. aureus, 40% of isolates were resistant to oxacillin, as were 63.4% of coagulase-negative staphylococcus isolates. Of 37 Gram-negative isolates, 35% were resistant to cefotaxime, 31% to ceftazidime, and 27% to ciprofloxacin. Imipenem and cefepime had the lowest reported rates of resistance (11%). PMID- 15355417 TI - Clinical-epidemiological characteristics and outcome of patients with catheter related bloodstream infections in Europe (ESGNI-006 Study). AB - This study analysed 89 episodes of catheter-related bloodstream infection (CR BSI) occurring during one week in 107 hospitals from 21 European countries (1.02 episodes/1,000 admissions). Patients from European Union (EU) countries had a higher incidence of CR-BSI than patients from non-EU countries (1.55 vs. 0.33/1,000 admissions). Most (67%) catheters were non-tunneled central venous catheters, were in the jugular vein (44%), had been implanted for > 7 days (70%), were made of polyurethane (61%) and were multi-lumen (67%). In 36% of cases, catheters were implanted by physicians other than anaesthetists or surgeons, and 50% were inserted by junior staff. PMID- 15355418 TI - Outbreak in France of Neisseria meningitidis B:15:P1.12 belonging to sequence type 1403. AB - This report describes a meningococcal outbreak in France caused by Neisseria meningitidis B:15:P1.12 of sequence type 1403, which affected eight young patients, between November 2000 and February 2002. Epidemiological typing confirmed that a single strain was responsible. Favourable outcome, sequelae or death resulted in similar proportions as in other cases of meningococcal disease in France during the same period, but purpura was observed in all eight cases. The patients were aged between 14 and 28 years, whereas the median age of patients affected by other meningococcal strains during this period in the same area was 60.4 years. PMID- 15355419 TI - Identification of Neisseria meningitidis sequence type 66 in Poland. AB - Investigation of two cases of invasive meningococcal disease within a single family revealed the presence of isolates of Neisseria meningitidis phenotype C:2b:P1.2,P1.5 belonging to sequence type (ST) 66. The ST66 clone is a single locus variant of the widely distributed ST8 complex, which has been observed previously in Spain, Belgium, Australia and New Zealand. This hypervariable meningococcal lineage has been responsible for local epidemics worldwide. This is the first report of ST66 meningococcal isolates of this phenotype from Poland. PMID- 15355420 TI - Increase of resistance to macrolides in invasive Streptococcus pneumoniae in Spain (2000-2001). AB - This study examined the antimicrobial resistance of 1,278 invasive Streptococcus pneumoniae isolates from 41 Spanish laboratories participating in the European Antimicrobial Resistance Surveillance System (EARSS) during 2000 and 2001. Twenty nine laboratories participated during both years and provided 950 of the isolates. Each laboratory used its own susceptibility testing methods. External quality assessment was performed annually by each participating laboratory. Significant increases in penicillin and erythromycin resistance were observed between 2000 and 2001. This increase was particularly noticeable in isolates from the laboratories participating during both years and in isolates from children and elderly patients. PMID- 15355421 TI - Acute and chronic otitis media and Turicella otitidis: a controversial association. AB - Turicella otitidis is a non-fermenting Gram-positive bacillus isolated almost exclusively from ear exudates. Its significance in acute or chronic otitis media is controversial. Over a 12-month period, T. otitidis was isolated from nine ear exudates from seven patients. Most of these were cases of spontaneous drainage following recurrence of otitis media after antimicrobial therapy that was ineffective against T. otitidis. The MICs of penicillin, levofloxacin, linezolid and vancomycin were very low for all the isolates studied, but most isolates displayed high resistance to macrolides and lincosamides. PMID- 15355422 TI - Commercial broth microdilution panel validation and reproducibility trials for NVP PDF-713 (LBM 415), a novel inhibitor of bacterial peptide deformylase. AB - NVP PDF-713 (LBM 415) is a peptide deformylase inhibitor being progressed into clinical trials. Dry-form broth microdilution panels of NVP PDF-713 were compared to reference MIC panels of 552 recent clinical isolates. Most (99.2%) dry-form MIC results were within +/- 1 log(2) dilution of the reference panel MICs. Of the bacteria tested, Streptococcus pneumoniae and Haemophilus influenzae showed a bias towards higher and lower MICs, respectively. Same-day and between-day reproducibility tests showed that 98.9% and 96.7% of MIC values, respectively, were within +/- 1 log(2) dilution step, thereby demonstrating a high degree of reliability of the dry-form MIC product for clinical studies. PMID- 15355423 TI - Absence of carriage of glycopeptide-resistant enterococci by at-risk hospitalised patients in Malta. PMID- 15355424 TI - Zygomycosis in neutropenic patients with past Aspergillus infection: a role for posaconazole? PMID- 15355425 TI - T-box genes and congenital heart/limb malformations. AB - Congenital malformations cause significant morbidity and mortality; however, the underlying basis for many of these developmental defects is not well understood. Over the past years, a new family of genes called T-box genes has been identified that play essential roles during the development of various tissues and organs. A number of developmental syndromes have recently been shown to be linked to mutations in T-box genes, and brought direct medical relevance to their study. This review emphasizes emerging data on the molecular, cell, and disease levels, which establish a basis for parallel events in limb and heart development, and suggests that common regulatory pathways are crucial for proper differentiation and growth of these embryonic structures. PMID- 15355426 TI - Will the new cytogenetics replace the old cytogenetics? AB - With the advent of array-based comparative genomic hybridization technology, the analog cytogenetic analysis that has been used for the past 100 years could be replaced by the quantitative, microarray-based molecular analysis. Major advantages of the new array-based cytogenetic technologies are the high resolution and the high throughput. This technology is the first to offer an autonomous whole-chromosome analysis in one hybridization reaction for the detection of submicroscopic gains/losses. However, as with any new technology, it needs to be validated with regard to its performance in various applications (e.g. clinical genetic testing and cancer applications), comparative cost, and the data interpretation. PMID- 15355427 TI - Mechanisms regulating the development of the corpus callosum and its agenesis in mouse and human. AB - The development of the corpus callosum depends on a large number of different cellular and molecular mechanisms. These include the formation of midline glial populations, and the expression of specific molecules required to guide callosal axons as they cross the midline. An additional mechanism used by callosal axons from neurons in the neocortex is to grow within the pathway formed by pioneering axons derived from neurons in the cingulate cortex. Data in humans and in mice suggest the possibility that different mechanisms may regulate the development of the corpus callosum across its rostrocaudal and dorsoventral axes. The complex developmental processes required for formation of the corpus callosum may provide some insight into why such a large number of human congenital syndromes are associated with agenesis of this structure. PMID- 15355428 TI - Genetic landmarks through philately--epilepsy and clinical genetic issues. AB - Epilepsy is a common neurologic disorder. Major advances in the understanding of the etiology and treatment have occurred. Although most cases of epilepsy do not follow a simple pattern of inheritance, recently single gene epilepsy disorders have been identified. We present some postage stamps to illustrate issues and advances in knowledge about epilepsy, as well as famous people with this disorder. PMID- 15355432 TI - Genetic variation at the perilipin (PLIN) locus is associated with obesity related phenotypes in White women. AB - Perilipin coats intracellular lipid droplets and modulates adipocyte lipolysis. We have evaluated the association between several polymorphisms at the perilipin (PLIN) locus (PLIN1 : 6209T > C, PLIN4 : 11482G > A, PLIN5 : 13041A > G, and PLIN6 : 14995A > T) with obesity-related phenotypes in 1589 White subjects randomly selected from a general Spanish population. In women (n = 801), the less common alleles of PLIN1 and PLIN4, in strong linkage disequilibrium (D' : 0.96), were significantly associated with lower body mass index. Carriers of the allele 2 (6209C) at the PLIN1 locus weighed significantly less (-2.2 kg; p = 0.007) than women homozygotes for the wild-type genotype. The same was true for 11482A carriers at PLIN4 (p = 0.01). Moreover, the PLIN4 variant was associated with significantly lower waist-to-hip ratio, plasma glucose, and triacylglycerol concentrations. No significant associations with these obesity-related phenotypes were found in men. In agreement with these results, statistically significant gene-gender interactions were obtained when the risk of obesity was estimated (281 subjects were obese and 1308 non-obese). Only in women, PLIN1 and PLIN4 variant alleles (6209C and 11482A) were associated with a lower obesity risk [Odds ratio (OR) = 0.58, 95% confidence interval (CI): 0.38-0.93 and OR = 0.56, 95% CI: 0.36-0.89, respectively]. In summary, our data suggest that common alleles at the PLIN locus modulate body weight and metabolic variables in humans. PMID- 15355433 TI - Genetic discrimination: the clinician perspective. AB - Clinicians attending continuing education sessions in California were surveyed about their beliefs and attitudes regarding genetic discrimination and their knowledge of protective legislation. Two hundred seventy-one surveys were collected from physicians (n = 191) and nurses (n = 80). Most respondents lacked information or were misinformed about the existence of protective legislation (58.3%) or published cases of insurance discrimination (85.2%); 52.4% believed that mutation carriers have difficulty obtaining health insurance; 13% would not encourage genetic testing, despite a family history of cancer. Clinician concerns about potential genetic discrimination, and lack of information regarding protective legislation, may influence access to care. PMID- 15355434 TI - Genotype-phenotype studies in three families with mutations in the polyglutamine binding protein 1 gene (PQBP1). AB - Recently, the polyglutamine-binding protein 1 (PQBP1) gene was found to be mutated in five of 29 families studied with X-linked mental retardation (XLMR) linked to Xp. The reported mutations include duplications or deletions of AG dinucleotides in the fourth coding exon that resulted in shifts of the open reading frame. Three of the five families with mutations in this newly identified XLMR gene have been reported previously. We characterized the phenotypic and neuropsychological features in the two unpublished families with aberrations in PQBP1 and in a family reported 10 years ago. In total, seven patients diagnosed with aberrations in this gene were examined, including a newly identified patient at 18 months of age. Additionally, the features were compared to those reported in the literature of three other families, comprising MRXS3 (Sutherland-Haan syndrome) MRX55 and MRXS8 (Renpenning syndrome). Characteristics seen in these patients are microcephaly, lean body habitus, short stature, striking facial appearance with long narrow faces, upward slant of the eyes, malar hypoplasia, prognathism, high-arched palate and nasal speech. In addition, small testes and midline defects as anal atresia or imperforate anus, clefting of palate and/or uvula, iris coloboma and Tetralogy of Fallot are seen in several patients. These observations contribute to the phenotypic knowledge of patients with PQBP1 mutations and make this XLMR syndrome well recognizable to clinicians. PMID- 15355435 TI - The dynamics of X-inactivation skewing as women age. AB - Non-random X-chromosome inactivation (XCI) has been associated with X-linked diseases, neoplastic diseases, recurrent pregnancy loss, and trisomy risk. It also occurs more commonly in older female populations. To understand the etiology of non-random XCI and utilize this assay appropriately in clinical research and practice, the age-related alteration in XCI patterns in normal females needs to be clearly defined. In the present study, we evaluated the XCI status in 350 unselected women aged 0-88 years with unknown history of genetic disorders or abnormal pregnancies. DNA samples were extracted from peripheral blood and analyzed by a methylation-based assay at the androgen receptor locus. A weak but significant positive correlation was observed between age and degree of skewing in XCI over the whole age range (r = 0.23, p < 0.0001), and skewing values become non-normally distributed at older ages. However, the increase in skewed XCI appears to be more pronounced after age 30 than at younger ages. This trend supports the model of increased skewing with age as a consequence of hematopoietic stem cell senescence. An alternative possibility is that there is allele-specific loss of methylation with time that results in the appearance of increased XCI skewing using a methylation-based assay. PMID- 15355436 TI - Screening of SLC26A4 (PDS) gene in Pendred's syndrome: a large spectrum of mutations in France and phenotypic heterogeneity. AB - Sensorineural hearing defect and goiter are common features of Pendred's syndrome. The clinical diagnosis of Pendred's syndrome remains difficult because of the lack of sensitivity and specificity of the thyroid signs. The identification of PDS as the causative gene allowed molecular screening and enabled a re-evaluation of the syndrome to identify potential diagnostic characteristics. This report presents the clinical and genotypic findings of 30 French families, for whom a diagnosis of Pendred's syndrome had been made. Twenty seven families had at least one mutated allele. Twenty-eight different mutations were identified, 11 of which had never been previously reported. The main clinical characteristics were: early hearing loss, fluctuation in terms of during deafness evolution, and the presence of an enlarged vestibular aqueduct. PMID- 15355437 TI - Genetic analysis of primary microcephaly in Indian families: novel ASPM mutations. AB - Patients with primary microcephaly, an autosomal recessive trait, have mild to severe mental retardation without any other neurological deficits. It is a genetically heterogeneous disorder with six known loci: MCPH1 to MCPH6. Only the genes for MCPH1 and MCPH5 have been identified so far. We have ascertained nine consanguineous families with primary microcephaly from India. To establish linkage of these nine families to known MCPH loci, microsatellite markers were selected from the candidate regions of each of the six known MCPH loci and used to genotype the families. The results were suggestive of linkage of three families to the MCPH5 locus and one family to the MCPH2 locus. The remaining five families were not linked to any of the known loci. DNA-sequence analysis identified one known (Arg117X) and two novel (Trp1326X and Gln3060X) mutations in the three MCPH5-linked families in a homozygous state. Three novel normal population variants (i.e., c.7605G > A, c.4449G > A, and c.5961 A > G) were also detected in the ASPM gene. PMID- 15355438 TI - Cost analysis of DNA-based testing in a large Canadian family with multiple endocrine neoplasia type 2. AB - One of the major goals of genetic testing is the reduction of morbidity and mortality. Given the appropriate circumstances, this can result in reduction in health care costs. Such savings can be demonstrated most effectively in large families with mutations in well characterized, dominantly acting genes. In our large family, a point mutation TGC>CGC in exon 10 of the RET proto-oncogene, which results in a missense mutation (Cys620Arg), was identified in two individuals. The proband has medullary thyroid carcinoma (MTC), as did her deceased mother. One son has MTC and Hirschsprung's disease. The proband's mother had nine siblings; the proband has three siblings, another son, and 69 maternal cousins. Genetic testing has been performed on the closest relatives and has identified four individuals with, and 54 individuals without, a familial RET mutation. Significant cost savings have been realized in both genetic testing and clinical surveillance. In this family, for every at-risk individual identified as a true-negative, the minimum yearly savings in clinical surveillance is 508 dollars per person. As demonstrated by this case, economic costs of genetic diagnostics should take into account the potential saved monies in tests, both molecular and clinical. PMID- 15355439 TI - Haplotype analysis at the ETM2 locus in a Singaporean sample with familial essential tremor. AB - An ancestral haplotype on chromosome 2p24.1 described in an American sample with familial essential tremor (ET) was analyzed in a different ethnic sample from Singapore. Six polymorphic loci (etm1240, etm1231, etm1234, APOB, etm1241, and etm1242) in a 274-kb interval within an ET gene candidate region (ETM2) were analyzed in Singaporean individuals with a family history of ET (n = 52) and compared to Singaporean controls older than age 65 (n = 49). The allele frequencies were significantly different between cases and controls for the loci etm1234 (p = 0.0001) and APOB (p = 0.0320). An extended haplotype formed by the loci etm1231, etm1234, and APOB occurred with a frequency of 31% in Singaporean cases and in 1.8% of elderly Singaporean controls (p = 0.0005). Haplotype studies in two different population samples suggest that a disease locus for ET lies near or within the 100-kb interval between the loci etm1231 and APOB. PMID- 15355440 TI - Refined mapping of the autosomal recessive non-syndromic deafness locus DFNB13 using eight novel microsatellite markers. AB - The locus for a type of an autosomal recessive non-syndromic deafness (ARND), DFNB13, was previously mapped to a 17-cm interval of chromosome 7q34-36. We identified two consanguineous Tunisian families with severe to profound ARND. Linkage analyses with microsatellites surrounding the previously identified loci detected linkage with markers corresponding to the DFNB13 locus in both families. Haplotype analyses assigned this locus to a 3.2-Mb region between markers D7S2468 and D7S2473. In order to refine this interval, we identified nine dinucleotide repeats in the 7q34 region. To investigate the polymorphism of these repeats, a population study of 74 unrelated individuals from different regions of Tunisia was carried out. Our results demonstrated that eight of the nine repeats are polymorphic. The average number of alleles at these informative loci was 9.12 with a polymorphism information content of 0.71. Little evidence for linkage disequilibrium between some marker pairs was found. Haplotype analysis using these microsatellites refined the DFNB13 interval to an area of 2.2 Mb between the D7S5377 and D7S2473. In order to identify the DFNB13 gene, we sequenced and eliminated three candidate genes. Other known and predicted genes are being screened for deafness-causing mutations. PMID- 15355441 TI - Common variants in beta2- and beta3-adrenergic receptor genes and uncoupling protein 1 as predictors of the risk for type 2 diabetes and body weight changes. The Finnish Diabetes Prevention Study. PMID- 15355442 TI - Genetic basis of Prader-Willi syndrome in Korea: less uniparental disomy than has been recognized? PMID- 15355444 TI - The thyroid and the skeleton. PMID- 15355445 TI - Medullary thyroid carcinoma. AB - Medullary thyroid carcinoma (MTC) arises from parafollicular or C cells that produce calcitonin (CT), and accounts for 5-10% of all thyroid cancers. MTC is hereditary in about 25% of cases. The discovery of a MTC in a patient has several implications: disease extent should be evaluated, phaeochromocytoma and hyperparathyroidism should be screened for and whether the MTC is sporadic or hereditary should be determined by a direct analysis of the RET proto-oncogene. In this review, pathological characteristics, tumour markers and genetic abnormalities in MTC are discussed. The diagnostic and therapeutic modalities applied to patients with clinical MTC and those identified with preclinical disease through familial screening are also described. Progresses concerning genetics, initial treatment, follow-up, screening and treatment of pheochromocytoma have permitted an improvement in the long-term outcome. However, there is no effective treatment for distant metastases, and new therapeutic modalities are urgently needed. PMID- 15355446 TI - Aberrant adrenal sensitivity to multiple ligands in unilateral incidentaloma with subclinical autonomous cortisol hypersecretion: a prospective clinical study. AB - BACKGROUND: Incidentally discovered adrenal tumours are frequently associated with subclinical autonomous cortisol hypersecretion of unknown origin. Aberrant hormone receptors have been observed in case reports of overt Cushing's syndrome. The question arises as to whether such receptors may be present in the functioning adrenal incidentaloma, which is common and might be a subclinical stage of Cushing's syndrome. PATIENTS AND METHODS: Twenty-one consecutive patients with a unilateral incidentaloma, the biochemical features of subclinical cortisol hypersecretion and/or the scintigraphic features of an autonomously functioning adrenal adenoma were investigated for plasma cortisol responses to various stimuli: upright posture, meal, combined hypothalamic-hormones, the vasopressin analogue terlipressin, glucagon, angiotensin II, the serotonin 5-HT4 agonist cisapride, and ACTH. Six normal controls were similarly investigated. All subjects were studied during 8 mg per day dexamethasone in order to avoid any ACTH-dependent variation of plasma cortisol. RESULTS: The most constant responses in adrenal incidentalomas were observed after stimulation by terlipressin (18/20 patients, 28-415% cortisol increase) and cisapride (17/21 patients, 25-364% cortisol increase). Eighteen out of 21 patients responded to several stimuli (cortisol increase >or= 25%), and all responded to at least one stimulus other than ACTH, while such responses were absent in the controls. Plasma ACTH remained suppressed in all subjects throughout the study. CONCLUSIONS: Aberrant membrane receptors detected by in vivo stimulation tests appear to be common in autonomously functioning unilateral adrenocortical adenomas. These receptors may be involved in the modulation of cortisol secretion in adrenal incidentaloma, with potential therapeutic consequences for the control of subclinical cortisol hypersecretion. PMID- 15355447 TI - Traumatic brain injury and subarachnoid haemorrhage are conditions at high risk for hypopituitarism: screening study at 3 months after the brain injury. AB - OBJECTIVE: Acquired hypopituitarism in adults is obviously suspected in patients with primary hypothalamic-pituitary diseases, particularly after neurosurgery and/or radiotherapy. That brain injuries (BI) can cause hypopituitarism is commonly stated and has been recently emphasized but the management of BI patients does not routinely include neuroendocrine evaluations. AIM: To clarify the occurrence of hypopituitarism in patients after traumatic brain injury (TBI) or subarachnoid haemorrhage (SAH) 3 months after the BI. SUBJECTS AND METHODS: The occurrence of hypopituitarism in conscious patients after traumatic brain injury [TBI, n = 100, 31 women, 69 men; age 37.1 +/- 1.8 years; body mass index (BMI) 23.7 +/- 0.4 kg/m(2); Glasgow Coma Scale (GCS) 3-15] or subarachnoid haemorrhage [SAH, n = 40, 14 men, 26 wpmen, 51.0 +/- 2.0 years; 25.0 +/- 0.6 kg/m(2); Fisher's scale 1-4] was studied in a multicentre study 3 months after the BI. All patients underwent wide basal hormonal evaluation; the GH/IGF-I axis was evaluated by GHRH + arginine test and IGF-I measurement. RESULTS: In TBI patients, some degree of hypopituitarism was shown in 35%. Total, multiple and isolated deficits were present in 4, 6 and 25%, respectively. Diabetes insipidus was present in 4%. Secondary adrenal, thyroid and gonadal deficit was present in 8, 5 and 17%, respectively. Severe GH deficiency (GHD) was the most frequent pituitary defect (25%). In SAH patients, some degree of hypopituitarism was shown in 37.5%. Despite no total hypopituitarism, multiple and isolated deficits were present in 10 and 27.5%, respectively. Diabetes insipidus was present in 7.5%. Secondary adrenal, thyroid and gonadal deficit was present in 2.5, 7.5 and 12.5%, respectively. Severe GHD was the most frequent defect (25%). CONCLUSIONS: TBI and SAH are conditions associated with high risk of acquired hypopituitarism. The pituitary defect is often multiple and severe GHD is the most frequent defect. Thus neuroendocrine evaluations are always mandatory in patients after brain injuries. PMID- 15355448 TI - Regulation of glucocorticoid receptor gamma (GRgamma) by glucocorticoid receptor haplotype and glucocorticoid. AB - OBJECTIVE: To measure glucocorticoid receptor gamma (GRgamma) expression in transformed lymphocytes from individuals of known GR gene haplotype. Recently, a glucocorticoid receptor haplotype (GAT) has been described that associates with increased sensitivity to dexamethasone. As there is strong linkage disequilibrium across the gene, this haplotype is likely to extend through exon 3, altered splicing of which generates the GRgamma isoform, a splice variant with impaired transactivation activity. Therefore we proposed that the GR haplotype affects glucocorticoid sensitivity either by influencing GRgamma expression basally, or in response to Gc exposure. DESIGN: We have measured expression of GRgamma, using a validated RT-PCR assay in human B lymphoblast cells of known haplotype under basal conditions, and after dexamethasone treatment. PATIENTS: The A549 human lung cell line and normal volunteers, five with the GAT GR haplotype and three with the CGA haplotype. MEASUREMENTS: Relative expression of GRgamma compared to total GR mRNA. RESULTS: GRgamma made up 5-6% of all the GR transcripts. There was no effect of carriage of the GR gene GAT haplotype on this expression. There was no effect of dexamethasone on relative expression of GRgamma. CONCLUSIONS: We propose that the GRgamma isoform is a product of constitutive splicing, that it does not explain the GR haplotype association with altered glucocorticoid sensitivity, and is unlikely to play an important physiological role in affecting glucocorticoid sensitivity. As glucocorticoids do not affect GRgamma expression, relative to total GR, this splice variant is unlikely to influence glucocorticoid treatment response. PMID- 15355449 TI - Effect of dieting on plasma leptin, soluble leptin receptor, adiponectin and resistin levels in healthy volunteers. AB - OBJECTIVE: Recent findings demonstrating important effects of the adipokines on metabolism, energy homeostasis and body weight regulation have prompted research on the possible role of negative energy balance in altering adipocytokine regulation. The goal of this study was to evaluate the effects of a hypocaloric diet in healthy normal-weight volunteers. An additional goal was to help clarify the contribution of restricted caloric intake to altered plasma adipokine levels in the eating disorders anorexia nervosa and bulimia nervosa. DESIGN: Participants were studied before and after a 4-week reduced-calorie diet (1000 12000 kcal/day). patients Subjects included 15 healthy, normal-weight women (age 22 +/- 3 years). MEASUREMENTS: Plasma concentrations of leptin, soluble leptin receptor protein (sOB-R), adiponectin, resistin, thyroid hormones and beta hydroxybutyrate were determined following overnight fast before and after the 4 week reduced-calorie diet. RESULTS: Subjects lost a mean of 3.4 +/- 2.1 kg in response to the reduced-calorie diet. The weight loss phase was associated with a 60.3% decrease in plasma leptin levels (P < 0.001), a 43.5% increase in sOB-R levels (P < 0.002) and a 16.2% decrease in plasma adiponectin levels (P < 0.04). There was no significant change in plasma resistin levels. CONCLUSIONS: These results demonstrate that a modest decrease in energy intake sustained over several weeks may play an important role in altering levels of plasma leptin and sOB-R. The findings also provide preliminary evidence that, in contrast to previous results in obese subjects, caloric restriction with accompanying weight loss in healthy, normal-weight volunteers may lead to decreased circulating adiponectin levels. Additional studies will be needed to clarify the contribution of altered energy intake to abnormalities in cytokine levels in the eating disorders. PMID- 15355450 TI - Both intrauterine growth restriction and postnatal growth influence childhood serum concentrations of adiponectin. AB - OBJECTIVE: Insulin resistance has been linked to intrauterine growth restriction; adiponectin is a strong determinant of insulin sensitivity. We aimed at studying the contributions of birthweight and insulin sensitivity to circulating adiponectin in children born small for gestational age (SGA). DESIGN: Cross sectional, hospital-based study dealing with insulin sensitivity in SGA children. PATIENTS: Thirty-two prepubertal children born SGA (age 5.4 +/- 2.9 years) and 37 prepubertal children born appropriate for gestational age (AGA, age 5.9 +/- 3.0 years). MEASUREMENTS: Serum levels of fasting glucose, serum lipids, insulin (immunometric assay) and adiponectin concentrations (ELISA) were assessed, and insulin resistance (IR) and insulin secretion (beta-cell) were calculated by the homeostasis model of assessment (HOMA). RESULTS: SGA children had similar HOMA IR, HOMA-beta-cell and adiponectin concentrations than AGA children. However, in a separate analysis of subjects older than 3 years of age, SGA children showed higher HOMA-IR after adjusting for sex, age and body mass index (BMI) standard deviation score (SDS). Circulating adiponectin was higher in SGA children [adjusted means: 14.5 mg/l (95% CI 12.9-16.1) and 18.7 mg/l (95% CI 17.0-20.3) for AGA and SGA children, respectively; P < 0.0001]. Further analysis revealed that the group of overweight SGA (arbitrarily defined as being in the higher quartile for the BMI SDS distribution in the sample) had decreased serum concentrations of adiponectin, compared to lean SGA children [adjusted means: 12.9 mg/l (95% CI 9.3-16.5) vs. 19.0 (95% CI 16.8-21.3), respectively; P = 0.001]. In a multiple regression model, HOMA-IR and SGA status explained 35% and 15% of adiponectin variance, respectively. CONCLUSIONS: Prenatal growth restriction is associated with insulin resistance but relatively increased adiponectin concentrations, provided overweight does not ensue. The contributions of circulating adiponectin to the increased risks for developing insulin resistance and type-2 diabetes in formerly SGA subjects merit further studies. PMID- 15355451 TI - Metabolism of chylomicrons in patients with congenital lipoatrophic diabetes: a study with emulsion models of chylomicrons. AB - BACKGROUND: Lipoatrophic diabetes is characterized by the near absence of adipose tissue and the presence of insulin-resistant diabetes. Fasting hypertriglyceridaemia and increased postprandial lipidaemia are also present, but the metabolism of chylomicrons, the triglyceride-rich lipoproteins in the circulation that carry the dietary fats absorbed by the intestine, was not specifically investigated. Because both the activity of insulin-dependent lipoprotein lipase that catalyses the chylomicron lipolysis and the storage of the lipolysis products are affected in the disease, it is important to evaluate how those changes may ultimately affect the chylomicron lipolysis and removal of chylomicron remnants from the circulation. OBJECTIVE: The aim of the study was to evaluate the chylomicron intravascular metabolism in patients with lipoatrophic diabetes. PATIENTS: Six patients with lipoatrophic diabetes (four females, two males) aged 22.2 +/- 4.4 years, with body mass index (BMI) 21.6 +/- 3.6 kg/m(2), were compared with 12 healthy control subjects (seven females, five males) aged 24.3 +/- 2.1 years with BMI 22.5 +/- 2.7 kg/m(2). MEASUREMENTS: The plasma kinetics of intravenously injected chylomicron-like emulsions labelled with (3)H triglycerides ((3)H-TG) and with (14)C-cholesteryl esters ((14)C-CE) were determined, the former tracing the chylomicron lipolysis by lipoprotein lipase and the latter the removal of chylomicron remnants from the plasma. RESULTS: Triglyceride values (8.3 +/- 9.2 mmol/l) in the patients were higher (P < 0.005) and high density lipoprotein (HDL) cholesterol values (0.8 +/- 0.2 mmol/l) lower (P < 0.0005) than in controls (0.7 +/- 0.2 and 1.3 +/- 0.4 mmol/l, respectively) whereas total cholesterol, apoprotein B (apo B) and apo A1 were similar. The fractional clearance rate (FCR, in min(-1)) of (3)H-TG was 0.014 +/- 0.016 and the FCR of (14)C-CE was 0.008 +/- 0.012 in the patients and 0.046 +/- 0.024 and 0.024 +/- 0.012 in the controls, respectively (P < 0.05). Thus FCRs of both emulsion labels were markedly reduced in the patients, indicating that lipolysis and remnant removal were diminished. Diminished remnant removal may be due to either deficient lipolysis or deficient removal mechanisms. CONCLUSION: The metabolism of chylomicrons tested by the emulsion method is impaired in lipoatrophic diabetes. PMID- 15355452 TI - The relation of insulin, leptin and IGF-1 to birthweight in offspring of women with type 1 diabetes. AB - OBJECTIVE: Maternal diabetes is associated with excess foetal growth. We have assessed the influence of maternal diabetes on hormones associated with foetal growth and the relationship of these hormones to birthweight. DESIGN: Case control study. PATIENTS: Singleton offspring of mothers with type 1 diabetes (ODM, n = 140) and control mothers (Control, n = 49). MEASUREMENTS: Birthweight, cord blood insulin, proinsulin, 32-33 split proinsulin, leptin, IGF-1, IGFBP-3, cortisol. RESULTS: Maternal diabetes was associated with higher birthweight (ODM 3.80 +/- 0.69 kg; Control; 3.56 +/- 0.52 kg, P = 0.02) and marked increases in insulin (median [interquartile range]: ODM 110 [60-217] pmol/l; Control 22 [15 37] pmol/l; P < 0.0001) and leptin (ODM 32 [15-60] ng/ml; Control 9 [4-17] ng/ml; P < 0.0001) but no absolute difference in IGF-1 (ODM 7.9 [6.2-9.8] nmol/l, Control 7.5 [6.2-9.8] nmol/l, P = 0.24) or its principle binding protein IGFBP-3 (ODM 1.63 +/- 0.38 micro g/ml, Control 1.63 +/- 0.28 micro g/ml; P = 0.12). Individually, insulin, insulin propeptides, leptin, IGF-1 and IGFBP-3 were significantly (P < 0.05) correlated with birthweight (in ODM and Control). IGF-1 and leptin were positively related to birthweight independently of each other and insulin in both ODM and Control. By contrast, insulin showed independent relationships to birthweight in ODM (P < 0.0001) but not in Control (P = 0.4). CONCLUSIONS: Maternal diabetes is associated with marked elevation of insulin and leptin in cord blood of their offspring. Hormonal correlates of birthweight differ between ODM and Control with an independent relationship of insulin to birthweight observed only in ODM. PMID- 15355453 TI - Insulin-like growth factor-1 deficiency determines increased intima-media thickness at common carotid arteries in adult patients with growth hormone deficiency. AB - OBJECTIVE: To investigate the role of IGF-1 on intima-media thickness (IMT) at common carotid arteries by Doppler ultrasonography. SUBJECTS: Thirty-nine patients (17 women, 22 men, aged 25-70 years) with severe GH deficiency (GHD), 19 with normal and 20 with low IGF-1 levels, and 39 sex-, age- and body mass index (BMI)-matched healthy controls. RESULTS: Patients with GHD showed abnormalities in lipid profile, and increased fibrinogen levels, mean IMT (0.88 +/- 0.26 vs. 0.69 +/- 0.14 mm, P < 0.001), and systolic and diastolic peak velocity (P < 0.001) compared to controls. Eight patients (18%) and one control (2.1%, P = 0.04) had well-defined plaques. In controls, but not in patients with GHD, mean carotid IMT was correlated with age (r = 0.78, P < 0.001). In both controls (r = 0.82; P < 0.0001) and patients with GHD (r = -0.84, P < 0.0001), serum IGF-1 levels were inversely correlated with mean IMT at common carotid arteries. At the stepwise multiple regression, the variables most significantly related to IMT in GH-deficient patients were total cholesterol levels (t = 5.2, P < 0.001), followed by disease duration (t = 2.4, P = 0.02), while in controls the variables most significantly related to IMT were IGF-1 levels (t = -9.9, P < 0.001), followed by low density lipoprotein (LDL)-cholesterol levels (t = -2.3, P = 0.02). Compared to patients with normal IGF-1 levels, those with low IGF-1 levels had lower high density lipoprotein (HDL)-cholesterol levels (1.0 +/- 0.2 vs. 1.3 +/- 0.2 mmol/l, P = 0.0002), and higher glucose (54.3 +/- 6.1 vs. 48.9 +/- 5.9 mmol/l, P = 0.008), insulin (25.2 +/- 6.8 vs. 18.8 +/- 6.0 mUl/l, P = 0.004), total cholesterol (7.1 +/- 1.1 vs. 4.9 +/- 0.6 mmol/l, P < 0.0001), total/HDL cholesterol ratio (7.2 +/- 1.8 vs. 3.9 +/- 0.7, P < 0.0001), fibrinogen levels (319.8 +/- 56.9 vs. 241.8 +/- 53.0 mg/dl, P < 0.0001) and mean IMT at common carotid arteries (1.05 +/- 0.25 vs. 0.69 +/- 0.07 mm, P < 0.0001). Atherosclerotic plaques were found only in GH-deficient patients with low IGF-1 levels. CONCLUSIONS: GH-deficient patients have alterations in lipid profile with an increase in the total/HDL-cholesterol ratio, which is an index of increased cardiovascular risk, but only patients with IGF-1 deficiency have increased IMT. PMID- 15355454 TI - Weight-related dosing, timing and monitoring hydrocortisone replacement therapy in patients with adrenal insufficiency. AB - OBJECTIVE: The objective of this study was to examine the variables determining hydrocortisone (HC) disposition in patients with adrenal insufficiency and to develop practical protocols for individualized prescribing and monitoring of HC treatment. DESIGN AND PATIENTS: Serum cortisol profiles were measured in 20 cortisol-insufficient patients (09.00 h cortisol < 50 nmol/l) given oral HC as either a fixed or 'body surface area-adjusted' dose in the fasted or fed state. Endogenous cortisol levels were measured in healthy subjects. Pharmacokinetic analysis was performed using P-Pharm software, and computer simulations were used to assess the likely population distribution of the data. RESULTS: Body weight was the most important predictor of HC clearance. A fixed 10-mg HC dose overexposed patients to cortisol by 6.3%, whereas weight-adjusted dosing decreased interpatient variability in maximum cortisol concentration from 31 to 7%, decreased area under the curve (AUC) from 50 to 22% (P < 0.05), and reduced overexposure to < 5%. Food taken before HC delayed its absorption. Serum cortisol measured 4 h after HC predicted cortisol AUC (r(2) = 0.78; P < 0.001). CONCLUSIONS: We recommend weight-adjusted HC dosing, thrice daily before food, monitored with a single serum cortisol measurement using a nomogram. This regimen was prospectively examined in 40 cortisol-insufficient patients, 85% of whom opted to remain on the new thrice-daily treatment regimen. PMID- 15355455 TI - Gonadotrophin receptor blocking antibodies measured by the use of cell lines stably expressing human gonadotrophin receptors are not detectable in women with 46,XX premature ovarian failure. AB - BACKGROUND: Premature ovarian failure (POF) is defined by cessation of ovarian function after puberty and before the age of 40. The syndrome is characterized by amenorrhoea, oestrogen deficiency and elevated levels of gonadotrophins. Autoimmunity has been proposed as a mechanism for some cases of destruction or malfunction of ovarian follicles. POF is often associated with type I and type II polyglandular autoimmune syndromes. It has also been postulated that receptors such as the LH and FSH receptors might become targets for blocking antibodies and such antibodies could be a cause of ovarian failure. PATIENTS AND METHODS: Sixty nine patients with POF isolated or associated with other endocrine autoimmune diseases (autoimmune thyroid diseases, Addison's disease, type 1 diabetes mellitus, multiple sclerosis, myasthenia gravis) were studied. All the patients had secondary amenorrhoea. The patient group had a median age of 33.1 years (range 15-57). Ovarian failure had been diagnosed at a median age of 29 years (range 15-39). The median time since diagnosis was almost 1 year but in six patients gonadal insufficiency had appeared 10-30 years earlier. All had a normal chromosomal karyotype (46, XX). Patients with POF were characterized by duration of amenorrhoea > 1 year, with elevated FSH and LH levels and undetectable or low oestrogen levels. Cell lines stably expressing recombinant human LH (CHO-LHr) and FSH (CHO-FSHr) receptors were prepared and used to search for antibodies able to inhibit LH- or FSH-stimulated cAMP production. Immunoglobulins extracted from sera of patients with POF were incubated with CHO-LHr and CHO-FSHr in the presence of human recombinant CG and FSH, respectively. RESULTS AND CONCLUSIONS: None of the immunoglobulin G (IgG) preparations from patients with POF was able to inhibit the activity of the FSH- and CG-stimulated cAMP production. PMID- 15355456 TI - Type V phosphodiesterase inhibitor treatments for erectile dysfunction increase testosterone levels. AB - OBJECTIVE: Lack of sexual activity due to erectile dysfunction (ED) decreases testosterone (T) levels through a central effect on the hypothalamic-pituitary axis. In this paper we studied the effect of different type V phosphodiesterase (PDE5) inhibitor treatments for ED on the reversibility of this endocrine pattern. DESIGN: Open-label, retrospective study. PATIENTS: Seventy-four consecutive patients were treated on demand with sildenafil (Sild) (50 mg) and tadalafil (Tad) 20 mg. MEASUREMENTS: The success in sexual intercourse was recorded and total (tT) and free testosterone (fT) levels were studied before and after 3 months of treatment. RESULTS: Basal level of tT and fT were at the bottom of the normal range and LH levels were at the top of the high normal range. After treatments, this endocrine pattern was reversed in both groups. However, the T increase in Sild-treated patients was significantly lower than in those treated with Tad (4.7 +/- 2.7 vs. 5.1 +/- 0.9, P < 0.001). fT levels followed a directly proportional pattern, while the inverse was found when LH production was studied. The intercourse rate reflected this effect: in fact, the Sild group showed a 4.9 +/- 2.9/month full sexual intercourse rate while in the Tad group a significantly higher rate of sexual intercourse was found (6.9 +/- 4.6/month, P = 0.04). However, drug consumption was comparable between the groups (Sild 4.9 +/- 2.9 vs. Tad 4.4 +/- 2.8 pills/month, P = 0.72). CONCLUSIONS: As it is unlikely that the two drugs have a different direct effect on the pituitary-testis axis, this effect is probably due to the higher frequency of full sexual intercourse in the Tad-treated group, because of the drug's longer half-life. PMID- 15355457 TI - Recombinant GH replacement in hypopituitary adults improves endothelial cell function and reduces calculated absolute and relative coronary risk. AB - OBJECTIVE: Adult GH deficiency (GHD) is linked to endothelial dysfunction and vascular disease. We examined the effect of 12 months of GH therapy on endothelial function, C-reactive protein (CRP) and coronary risk. DESIGN: Open design intervention study. PATIENTS: Fourteen GH-deficient patients (nonsmokers, without diabetes, hypertension or vascular disease) studied before, 6 months and 12 months after GH therapy. MEASUREMENTS: Flow-mediated dilatation (FMD), carotid intima-media thickness (IMT) thrombomodulin (TM), E-selectin, CRP, lipid profile, blood pressure and anthropometric data were recorded. We used the Framingham equation to calculate coronary risk. RESULTS: FMD improved (7.5 +/- 1.62 vs. 11.93 +/- 1.52, P = 0.038). Overall there was no change in IMT, TM, E-selectin or CRP. The correlation between TM and FMD showed a trend for statistical significance (r = -0.54, P = 0.056). Changes in CRP correlated with change in IGF 1 (r = -0.67, P = 0.012); E-selectin correlated with high density lipoprotein (HDL)-cholesterol (r = -0.60, P = 0.028), triglycerides (r = 0.68, P = 0.01) and waist-to-hip ratio (WHR) (r = 0.71, P = 0.006). Systolic (127.36 +/- 4.47 vs. 120.36 +/- 3.50, P = 0.017) and diastolic (84.71 +/- 2.73 vs. 76.93 +/- 2.03, P = 0.005) blood pressure decreased. HDL-cholesterol increased (0.70 +/- 0.05 vs. 0.93 +/- 0.06, P = 0.001). WHR decreased (0.90 +/- 0.02 to 0.88 +/- 0.02, P = 0.043) without changes in weight or body mass index (BMI). Ten-year absolute (P = 0.009) and relative (P = 0.002) cardiac risk decreased. CONCLUSION: Biophysical test of endothelial function (FMD) improved after 12 months of GH therapy but there was no significant change in biochemical endothelial or inflammatory markers. Calculated coronary risk decreased mainly due to reduction in systolic and diastolic blood pressure and increase in HDL-cholesterol. PMID- 15355458 TI - Characteristics of recovery of adrenocortical function after treatment for Cushing's syndrome due to pituitary or adrenal adenomas. AB - OBJECTIVE: Surgical cure of Cushing's syndrome (CS) is followed by adrenocortical insufficiency, which may be long-lasting. The aim was to elucidate recovery of adrenocortical function, defined as a normal cortisol response to ACTH stimulation, and the relation to ACTH in patients cured for CS due to pituitary Cushing's disease (CD) or adrenal (AA) adenomas. DESIGN: A retrospective study including 32 patients considered surgically cured for CS (18 CD, 14 AA). RESULTS: Twelve (67%) patients with CD recovered within median 24 months (range 7 months 4(1)/(2) years) whereas six did not recover within 3-12 years. Plasma ACTH (p ACTH) at time of recovery was not different from p-ACTH in patients not recovering (P = 0.9). Eleven (79%) patients with AA recovered within 24 months (10 months-4 years) whereas three did not recover within 4-10 years. p-ACTH at time of recovery was higher compared to patients not recovering (P < 0.04). No differences were observed comparing CD and AA patients concerning preoperative 24 h urinary free cortisol (UFC) excretion, postoperative unstimulated s-cortisol or recovery time. By contrast, p-ACTH measured at time of recovery was higher in AA compared to CD (median 12.3 vs. 4.6 pmol/l) (P < 0.001), whereas plasma dehydroepiandrosterone sulfate (p-DHEAS) was lower in AA compared to CD (median 300 vs. 1500 nmol/l) (P = 0.02). CONCLUSION: Recovery of secondary adrenal insufficiency is a slow process in both CD and AA. ACTH measured at time of recovery was significantly higher and DHEAS significantly lower in patients with AA compared to CD, which may suggest different mechanisms of the recovery process and different set points in the glucocorticoid feedback inhibition of ACTH secretion. PMID- 15355459 TI - Changes in endothelium-dependent arterial dilation before and after subtotal thyroidectomy in subjects with hyperthyroidism. AB - OBJECTIVE: This case-control study was carried out to assess the alteration of endothelium-dependent arterial dilation before and after subtotal thyroidectomy in subjects with hyperthyroidism. PATIENTS AND METHODS: The study subjects included 12 patients with hyperthyroidism and 39 apparently healthy individuals. We performed a subtotal thyroidectomy on the hyperthyroid patients. The endothelium-dependent arterial dilation was determined with a high-resolution ultrasound method in each patient at the hyperthyroid stage before treatment (stage H), the euthyroid stage induced immediately before surgery (stage E), and the transient hypothyroid stage 1 or 2 months after surgery (stage L). RESULTS: The flow-mediated arterial dilation decreased significantly from H to E and from E to L (P < 0.001). As compared with H, baseline blood flow decreased markedly at stages E and L (P < 0.001). The flow-mediated arterial dilation and baseline blood flow in the control subjects were very close to those at stage E of the hyperthyroid patients. The absolute change in the flow-mediated arterial dilation showed significant negative correlation with the changes in TSH (r =-0.86, P < 0.001), lipoprotein (a) [Lp(a)] (r =-0.77, P < 0.001) and low density lipoprotein (LDL) (r =-0.79, P < 0.001), and significant positive correlation with changes in fT3 (r =+0.88, P < 0.001). The absolute change in the baseline blood flow showed significant positive correlation with the change in fT3 (r =+0.85, P < 0.001) and significant negative correlation with the change in TSH (r =-0.63, P < 0.01). CONCLUSION: The endothelium-dependent arterial dilation increases significantly in untreated hyperthyroid patients, and decreases markedly after a subtotal thyroidectomy. Therefore, we conclude that the endothelium is more responsive to reactive hyperaemia in the hyperthyroid than the euthyroid state. PMID- 15355460 TI - Lipoxins in asthma: potential therapeutic mediators on bronchial inflammation? AB - Arachidonic acid metabolism represents an important source of mediators with ambivalent actions. Among these, lipoxins (LXs) are the first agents identified and recognized as anti-inflammatory endogenous lipid mediators, which are involved in the resolution of inflammation and are present in the airways of asthmatic patients. Lipoxins result mainly from the interaction between 5 and 15 lipoxygenases (LO) and their levels are modulated by the degree of bronchial inflammation as well as by the long-term glucocorticoid treatments. In the airways, LX synthesis is higher in mild asthmatics than in severe asthmatics, whereas in vitro chemokine release inhibition by LXs is more effective in cells from severe asthmatics than from mild asthmatics. LipoxinA(4) effects on interleukin (IL)-8 released by blood mononuclear cells and on calcium influx in epithelial cells are mediated by the specific receptor ALX. Lipoxin generation by lung epithelial cells depends mainly on 15-LO activity. Mild asthmatics present higher 15-LOb expression at the epithelium level than severe patients, whereas the LX deficit in severe asthma is associated with an up-regulation of the 15-LOa expressions. Therefore, bronchial epithelial cells become a target for therapeutic intervention and LXs represent a potential therapeutic solution for bronchial inflammation resolution in asthma. PMID- 15355461 TI - Glucocorticoid receptors in human airways. AB - Inhaled and intranasal glucocorticoids are the most common and effective drugs for controlling symptoms and airway inflammation in respiratory diseases such as asthma, allergic rhinitis, and nasal polyposis. The last few years have seen a growing understanding of the mechanisms of glucocorticoid action and, in particular, the receptor that mediates glucocorticoid actions, the glucocorticoid receptor (GR). In this revision we present an update on the GR gene, the expression and regulation of its gene products, namely GRalpha and GRbeta, as well as their alterations in pathological states. GRalpha is responsible for the induction and repression of target genes, it is expressed in virtually all human cells and tissues, and its expression is known to be downregulated by glucocorticoids. GRbeta has been found to act as a dominant negative inhibitor of GRalpha-mediated transactivation in in vitro studies with transfected cells, but it does not appear to have a significant inhibitory effect on GRalpha-mediated transrepression. In addition, for most tissues the expression of GRbeta, at least at the mRNA level, is extremely low compared with that of GRalpha. Some pro inflammatory cytokines appear to upregulate the expression of GRbeta, and increased GRbeta expression has been reported in diseases associated with glucocorticoid resistance or insensitivity, such as bronchial asthma, nasal polyposis, and ulcerative colitis. However, the possible role of GRbeta in modulating glucocorticoid sensitivity and/or resistance in vivo has been highly debated and it is not yet clear. PMID- 15355462 TI - Maximal airway plateau responses and eosinophils in cough variant asthma. PMID- 15355463 TI - Sputum eosinophil counts and eosinophil cationic protein levels in cough-variant asthma and in classic asthma, and their relationships to airway hypersensitivity or maximal airway response to methacholine. AB - BACKGROUND: The aims of this study were to compare the degree of airway inflammation in cough-variant asthma (CVA) with that in classic asthma (CA), and to examine the relationship between airway inflammation and airway hypersensitivity or maximal airway response to methacholine in both conditions. METHODS: Sputum was induced in 41 CVA patients, in 41 methacholine PC(20)-matched CA patients, and in 20 healthy children. The sputum samples were analyzed for total and differential cell counts, and for eosinophilic cationic protein (ECP). A high-dose methacholine challenge test was performed in CVA and CA patients to determine PC(20) and maximal airway response. RESULTS: Sputum eosinophil percentages and ECP levels were significantly elevated in CVA and CA vs the control, but no significant differences were found between the two asthma groups. In the two asthma groups, neither sputum parameters correlated significantly with methacholine PC(20). However, the absence of a maximal response plateau or its higher level, when present, was associated with increased eosinophil percentages and ECP levels in the CVA group. CONCLUSIONS: The degree of eosinophilic inflammation may not be causally related to differences in presented asthma manifestations. The identification of a maximal response plateau and the level of this plateau in patients with CVA may provide information pertinent to airway eosinophilic inflammation. PMID- 15355464 TI - The association of hepatitis A and Helicobacter pylori with sensitization to common allergens, asthma and hay fever in a population of young British adults. AB - BACKGROUND: A negative association of oro-faecally spread infection with serological markers of sensitization and allergic disease has been reported. METHOD: Previous infection with hepatitis A and Helicobacter pylori was assessed in a community-based sample of young British adults and associations with serum specific IgE to environmental allergens, asthma-like symptoms and hay fever were examined. RESULTS: There was no association of previous infection with hepatitis A or H. pylori with wheeze or hay fever. There was no evidence of an association of infection with either agent and sensitization except for the isolated finding of a lower prevalence of sensitization to grass in those with IgG antibodies to H. pylori (OR 0.65, 95% CI 0.43-0.99). This association did not explain the negative association of family size with sensitization to grass. CONCLUSION: In this population, there was no evidence that infection with hepatitis A or H. pylori was associated with lower levels of IgE sensitization, asthma or hay fever except for an isolated finding of a negative association of H. pylori infection with sensitization to grass. PMID- 15355465 TI - Atopic disorders among Estonian schoolchildren in relation to tuberculin reactivity and the age at BCG vaccination. AB - BACKGROUND: Published data about a relationship of atopic diseases to Bacillus Calmette-Guerin (BCG) vaccination and tuberculin responses are inconsistent. Our aim was to determine this association in a country with a low prevalence of allergies. METHODS: A random sample of 10-11-year-old schoolchildren in Tallinn was studied by a parental questionnaire (n = 979) and skin-prick tests (n = 643), according to the International Study of Asthma and Allergies in Childhood. Data about BCG vaccinations and tuberculin tests were obtained from school records (n = 723). RESULTS: The prevalence of allergic symptoms and atopy was similar in children vaccinated during the first month of life and later. Positive tuberculin responses (> or =5 mm) were inversely related to symptoms of asthma [odds ratio (OR) 0.10 (95% confidence interval 0.00-0.68) for exercise-induced wheezing; OR 0.37 (0.12-0.99) for night cough], and eczema [OR 0.53 (0.28-0.98)] but not to atopy. However, among BCG-revaccinated children, atopy tended to be more common in tuberculin responders, and the atopic children were significantly more likely to have a positive tuberculin response after the revaccination than would be predicted by their first test. CONCLUSIONS: We found no protective effect of early BCG vaccination against atopy in school age, although tuberculin responses and allergic symptoms were inversely related. PMID- 15355466 TI - Bronchial hyperresponsiveness in children with atopic rhinitis: a 7-year follow up. AB - BACKGROUND: A high prevalence of bronchial hyperresponsiveness (BHR) was found in atopic subjects with rhinitis. Those subjects may be at higher risk for developing bronchial asthma. We evaluated, in a 7-year follow-up, BHR and atopy in a homogeneous population of nonasthmatic children with allergic rhinitis (AR), and their role in asthma development. METHODS: Twenty-eight children (6-15 years) with AR were studied. At enrollment (T(0)), skin tests, total serum IgE assay, peak expiratory flow (PEF) monitoring and methacholine (Mch) bronchial challenge were performed. BHR was computed as the Mch dose causing a 20% forced expiratory volume (FEV)(1) fall (PD(20)FEV(1)) and as dose-response slope (D(RS)). Subjects were reassessed after 7 years (T(1)) using the same criteria. RESULTS: At T(0), 13 children (46%), showing a PD(20)FEV(1) <1526 microg of Mch, had BHR (Mch+), although PEF variability (PEFv) was within normal limits. None of the children with negative methacholine test developed bronchial asthma after 7 years. Of the 13 Mch+, only two reported asthma symptoms after 7 years. No significant change was seen in the other parameters of atopy considered. CONCLUSION: Children with allergic rhinitis present a high prevalence of BHR. Nevertheless, their PEFv is normal and the rate of asthma development low. PMID- 15355467 TI - Allergen stimulates bone marrow CD34(+) cells to release IL-5 in vitro; a mechanism involved in eosinophilic inflammation? AB - The specific mechanisms that alter bone marrow (BM) eosinophilopoiesis in allergen-induced inflammation are poorly understood. The aims of this study were to evaluate (a) whether the number of BM CD34(+) cells is altered due to allergen sensitization and exposure in vivo and (b) whether BM CD34(+) cells produce and release interleukin (IL)-5, IL-3 and granulocyte macrophage-colony stimulating factor (GM-CSF) after stimulation in vitro. A mouse model of ovalbumin (OVA) induced airway inflammation was used. Bone marrow CD34(+) cells were cultured in vitro and the cytokine release was measured by enzyme-linked immunosorbent assay. The IL-5-production from CD34(+) cells was confirmed by immunocytochemistry. Airway allergen exposure increased the number of BM CD34(+) cells (P = 0.01). Bone marrow CD34(+) cells produced IL-5 when stimulated with the allergen OVA in vitro, but not IL-3 or GM-CSF. Nonspecific stimulus with calcium ionophore and phorbol-myristate-acetate of BM CD34(+) cells caused release of IL-5, IL-3 and GM CSF. The induced release of IL-5 was increased in alum-injected vs naive mice (P = 0.02), but was not affected by allergen sensitization and exposure. The release of IL-3 and GM-CSF was increased after allergen sensitization and exposure (P < 0.02). In conclusion, allergen can stimulate BM CD34(+) cells to produce IL-5 protein. It is likely that the CD34(+) cells have autocrine functions and thereby regulate the early stages of BM eosinophilopoiesis induced by airway allergen exposure. Alum, a commonly used adjuvant, enhances the release of IL-5 and may thereby enhance eosinophilopoiesis. PMID- 15355468 TI - Human mast cells express receptors for IL-3, IL-5 and GM-CSF; a partial map of receptors on human mast cells cultured in vitro. AB - BACKGROUND: Mast cells have long been recognized as the principal cell type that initiates the inflammatory response characteristic of acute allergic type 1 reactions. Our goal has been to further characterize maturation of progenitors to mast cells. METHODS: Mast cells were cultured from human cord blood derived CD133(+) progenitors. Mast cell function was tested using histamine release. During differentiation mast cells surface marker expression was monitored by flow cytometry. RESULTS: CD133(+) progenitors expressed the early haematopoietic and myeloid lineage markers CD34, CD117, CD13 and CD33. Mature mast cells expressed CD117, CD13 and CD33, and expression of the high affinity immunoglobulin E receptor FcepsilonRI increased during culture. Cytokine receptors interleukin (IL)-5R, IL-3R, granulocyte-macrophage-colony stimulating factor (GM-CSF)R and IL 18R were expressed at high levels during maturation. Chemokine receptors CXCR4 and CXCR2 were highly expressed on both newly purified CD133(+) cells and mature cells. CONCLUSION: Human mast cells can be cultured from a CD34(+)/CD117(+)/CD13(+)/CD33(+) progenitor cell population in cord blood that is tryptase and chymase negative. Developing and mature mast cells express a wide range of chemokine and cytokine receptors. We found high levels of expression of CD123, IL-5R and GM-CSF receptors, also found on eosinophils and basophils, and high levels of expression of the receptor for the inflammatory cytokine IL-18. PMID- 15355469 TI - Cat allergen peptide immunotherapy reduces CD4(+) T cell responses to cat allergen but does not alter suppression by CD4(+) CD25(+) T cells: a double-blind placebo-controlled study. AB - BACKGROUND: We have previously described both modification of allergen immunotherapy using peptide fragments, and reduced regulation of allergen stimulated T cells by CD4(+) CD25(+) T cells from allergic donors when compared with nonallergic controls. It has been suggested that allergen immunotherapy induces regulatory T cell activity: we hypothesized that allergen peptide immunotherapy might increase suppressive activity of CD4(+) CD25(+) T cells. OBJECTIVE: To examine cat allergen-stimulated CD4 T cell responses and their suppression by CD4(+) CD25(+) T cells before and after cat allergen peptide immunotherapy in a double-blind placebo-controlled study. METHODS: Peripheral blood was obtained and stored before and after peptide immunotherapy or placebo treatment. CD4(+) and CD4(+) CD25(+) were then isolated by immunomagnetic beads and cultured with allergen in vitro. RESULTS: Comparing cells from blood taken before with that after peptide immunotherapy there was a significant reduction in both proliferation and IL-13 production by allergen-stimulated CD4+ T cells, whereas no change was seen after placebo. CD4(+) CD25(+) T cells suppressed both proliferation and IL-13 production by CD4(+) CD25(-) T cells before and after therapy but peptide therapy was not associated with any change in suppressive activity of these cells. CONCLUSION: Allergen peptide immunotherapy alters T cell response to allergen through mechanisms other than changes in CD4(+) CD25(+) T cell suppression. PMID- 15355470 TI - The basophil activation test in wasp venom allergy: sensitivity, specificity and monitoring specific immunotherapy. AB - BACKGROUND: As in vitro diagnosis of wasp venom sensitization by specific serum IgE has a sensitivity of only 60-80%, additional in vitro tests are desirable. Basophil activation is associated with the expression of CD63 and its measurement has been proposed as a novel in vitro test for immediate-type allergy. Furthermore, to date, no in vitro test exists to monitor successful specific immunotherapy (SIT) with wasp venom. Therefore, the potentially harmful sting challenge is still recommended. OBJECTIVE: We compared the CD63-based basophil activation test (BAT) in the diagnosis of wasp venom allergy with skin tests and measurement of specific IgE. Furthermore, we investigated whether BAT can predict the outcome of the sting challenge in patients on SIT. METHODS: Fifty patients with a systemic reaction caused by a wasp sting and 20 controls were studied. Intracutaneous tests were performed with wasp and bee venom in the suspected allergics. Specific IgE was determined by the CAP-FEIA method and basophil activation by flow cytometry upon double staining with anti-IgE/anti-CD63 mAb. Twenty-five patients were sting challenged 6 months after starting SIT and the BAT was repeated before challenge. RESULTS: Sensitivity of the intracutaneous tests, specific IgE and BAT was 100, 76, and 92%, respectively. Specificity of specific IgE and the BAT was 85 and 80%, respectively. The cut-off for a positive BAT was 15% CD63+ basophils. There was a positive correlation between IgE reactivity to wasp venom and the number of CD63+ basophils (r = 0.65). Although no patient had a systemic reaction upon sting challenge, in most subjects basophil activation did not decrease when compared with the BAT before SIT. CONCLUSIONS: Quantitation of basophil activation by CD63 expression is a valuable new in vitro method for diagnosis of allergy to hymenopteran venoms. The CD63 based BAT is a helpful tool for the complementation of routine diagnostic tests such as specific IgE as it increases sensitivity of in vitro detection of sensitization. However, this in vitro method does not offer an alternative to the sting challenge in monitoring successful SIT. PMID- 15355471 TI - The CD63 basophil activation test in Hymenoptera venom allergy: a prospective study. AB - BACKGROUND: The basophil activation test (BAT), which relies on flow cytometric quantitation of the allergen-induced up-regulation of the granule-associated marker CD63 in peripheral blood basophils, has been suggested to be a useful approach in detecting responsiveness to allergens. The purpose of this study was to establish the usefulness of the BAT with regard to the clinical history and current diagnostic tools in Hymenoptera venom allergy using a prospective study design. METHODS: Fifty-seven consecutive patients allergic to Hymenoptera venom as defined by a systemic reaction after an insect sting, and 30 age- and sex matched control subjects with a negative history were included. The degree and nature of sensitization was confirmed by skin testing, specific immunoglobulin E (IgE), serum tryptase levels and BAT. In the nonallergic control group only analysis of specific IgE and BAT were performed. Correlation of BAT, skin test and specific IgE, respectively, with the clinical history in the allergic group was termed as sensitivity and in the control group as specificity. RESULTS: Twenty one of 23 (91.3%) bee venom allergic patients and 29 of 34 (85.3%) patients allergic to wasp and hornet venom tested positive in BAT. The overall sensitivity of BAT, specific IgE and skin tests were 87.7, 91.2 and 93.0%, respectively. The overall specificities were 86.7% for BAT and 66.7% for specific IgE. No correlation between the severity of clinical symptoms and the magnitude of basophil activation was observed. CONCLUSION: The BAT seems to be an appropriate method to identify patients allergic to bee or wasp venom with a comparable sensitivity to standard diagnostic regimens. The higher specificity of BAT as compared with specific IgE makes this test a useful tool in the diagnosis of Hymenoptera venom allergy. PMID- 15355472 TI - Pseudoallergic reactions in chronic urticaria are associated with altered gastroduodenal permeability. AB - BACKGROUND: In a subgroup of patients with chronic urticaria (CU) the disease is caused by pseudoallergic reactions to food. The aim of this study was to investigate whether disturbances of the gastrointestinal barrier function play a role in the pathomechanism of the disease. METHODS: In 55 patients with CU gastrointestinal permeability was measured with an in vivo triple-sugar-test before and after 24 days of a diet low in pseudoallergens. Sucrose served as marker for gastroduodenal permeability, lactulose/mannitol ratio for intestinal permeability. RESULTS: Basal gastroduodenal and intestinal permeability were significantly higher in patients with urticaria as compared to controls. In 29 of the 55 patients skin symptoms decreased or completely disappeared during the diet (responders). Compared to nonresponders (n = 26), responders had a significantly higher gastroduodenal permeability before treatment (0.36 +/- 0.04 vs 0.15 +/- 0.01% sucrose; P < 0.001), which decreased after the diet (0.17 +/- 0.02; P < 0.001). The number of patients with Helicobacter pylori infections did not differ between the two groups. CONCLUSIONS: The results indicate that in a subgroup of patients with CU and pseudoallergy an impaired gastroduodenal barrier function may be of pathophysiological importance. The underlying mechanisms seem to be independent of H. pylori infection. PMID- 15355473 TI - IVIG effects on autoantibody elimination. PMID- 15355475 TI - Lower thresholds for bronchial challenge testing. PMID- 15355476 TI - Asthma induced by latex from 'Christmas flower' (Euphorbia pulcherrima). PMID- 15355477 TI - Hypersensitivity pneumonitis caused by triglycidyl isocyanurate. PMID- 15355478 TI - Cyclosporine treatment decreases the percentage of cutaneous lymphocyte antigen (CLA)(+)CD4(+) T cells in children with severe atopic dermatitis. PMID- 15355479 TI - The accuracy of the diagnosis of suspected macrolide antibiotic hypersensitivity: results of a single-blinded trial. PMID- 15355480 TI - Partial safety of the new COX-2 inhibitor rofecoxib in NSAIDs high sensitive patients. PMID- 15355481 TI - Anaphylaxis due to ribostamycin. PMID- 15355482 TI - Allergy to wine. PMID- 15355483 TI - A case of pollinosis to Broussonetia papyrifera. PMID- 15355484 TI - The psychiatric comorbidity of epilepsy. AB - Several studies have assessed the prevalence of psychiatric disorders in epilepsy. They are characterized by considerable heterogeneity, because of differences in the population setting and type of study. A non-systematic review of the literature allows us to draw some useful, although not definite, conclusions. Six per cent of people with epilepsy in the general population appear to suffer from a psychiatric disorder, while this rises to 10-20% in populations with temporal lobe and/or refractory epilepsy. Mood disorders are the most common culprit (24-74%), particularly depression (30%), followed by anxiety disorders (10-25%), psychoses (2-7%) and personality disorders (1-2%). This comorbidity appears to be related to endogenous and exogenous (including iatrogenic) factors and to the severity and chronicity of epilepsy. Conditions such as schizophrenia-like psychosis of epilepsy and interictal dysphoric disorder are represented only in epilepsy. Adequate recognition and treatment of psychiatric conditions in epilepsy is essential for patient management because of their considerable burden in morbidity and quality of life. PMID- 15355485 TI - Topiramate in painful diabetic polyneuropathy: findings from three double-blind placebo-controlled trials. AB - OBJECTIVES: To evaluate the efficacy and tolerability of topiramate in patients with painful diabetic polyneuropathy. MATERIALS AND METHODS: Patients with moderate to extreme pain (0-4 Categorical Pain Scale score > or = 2) were randomized to placebo or topiramate (100, 200, or 400 mg/day) in three similar double-blind trials. The primary efficacy analysis was pain reduction from final visit to baseline in the 100-mm Visual Analog Scale (VAS) for the intent-to-treat populations. RESULTS: After 18-22 weeks of double-blind treatment, pain reductions were numerically greater with topiramate in two studies but differences between topiramate and placebo in VAS scores or in the secondary efficacy endpoints did not reach statistical significance in any of the three studies. Across all studies, 24% of topiramate-treated patients and 8% of placebo treated patients discontinued due to adverse events; groups did not differ in the occurrence of serious adverse events. CONCLUSION: These studies did not find topiramate to be significantly more effective than placebo in reducing pain scores in patients with painful diabetic polyneuropathy. Several design features may have precluded the studies from having sufficient sensitivity to differentiate effective and ineffective treatments. The study design and results are instructive for other investigators designing future clinical studies in neuropathic pain. PMID- 15355486 TI - Intraoperative human vagus nerve compound action potentials. AB - OBJECTIVE: Although electrical stimulation of vagus nerve is used widely for treatment of epilepsy the electrophysiological properties of human vagus nerve are not well characterized. Our objective was to measure compound action potentials of human vagus nerve fiber groups intraoperatively by stimulation using a commercially available generator and electrode system (Neurocybernetic Prosthesis System, NCP). MATERIAL AND METHODS: During NCP implantation we recorded compound action potentials evoked by stimulating the left vagus nerve through the NCP bipolar lead. Current intensities were varied from 0.25 to 3.0 mA. RESULTS: Vagus nerve compound action potential components conducting in the A, Adelta, and C velocity ranges could be elicited using either the NCP pulse generator or by a standard evoked potential instrument. A fiber potentials were recordable in all nerves, and were activated by very low stimulus currents. Adelta and C fibers were less reliably elicited, with C fibers requiring the highest currents. CONCLUSIONS: Three clearly identified fiber populations can be identified using therapeutic electrical stimulation of the human vagus. Intraoperative measurements of NCP-induced action potentials may potentially provide a marker for therapeutic stimulation and better insight into mechanisms of vagus nerve stimulation (VNS) efficacy. PMID- 15355487 TI - IgA antibodies against gliadin and gluten in multiple sclerosis. AB - BACKGROUND: Multiple changes in antibodies against various antigens are found in multiple sclerosis (MS). OBJECTIVE: We wanted to measure immunoglobulin A (IgA) antibodies to some common food antigens in MS and also IgG against gliadin and gluten. METHODS: The IgA antibodies were measured in serum against gluten, gliadin, lactoglobulin, lactalbumin, casein and ovalbumin in patients with MS and controls using ELISA technique. IgG was likewise measured for gluten and gliadin. RESULTS: Highly significant increases compared with controls were found for IgA and IgG antibodies against gliadin and gluten. IgA antibodies against casein were significantly increased. Anti-endomycium and anti-transglutaminase antibodies were negative. CONCLUSIONS: The data presented indicate that there may be a possible moderately increased uptake of some specific proteins from the gut in MS compared with controls. PMID- 15355488 TI - Serial changes in fibrinolysis and coagulation activation markers in acute and convalescent phase of ischemic stroke. AB - BACKGROUND: The changes in the activity of a number of plasma markers of coagulation and fibrinolysis have previously been studied in patients with ischemic stroke, with conflicting results. We aimed to find out the changes in the activities of a wide array of markers of the coagulation and the fibrinolytic system of mildly or moderately affected first-ever ischemic stroke patients. METHODS: In a prospective, longitudinal, case-control study, we studied plasma plasminogen activator inhibitor type-1 (PAI-1) activity, tissue-type plasminogen activator antigen (t-PA:Ag), d-dimer, prothrombin fragment 1+2 (F 1+2), and thrombin-antithrombin III complex (TAT) levels in 55 consecutive patients on admission, 1 week, 1 month, and 3 months after an ischemic stroke. Sex- and age matched controls were studied once. All patients underwent blood sampling at each study time point; comprehensive stroke risk factors were recorded, and the etiology of the ischemic stroke was determined. All patients were contacted 3 years later for possible recurrent ischemic events. RESULTS: PAI-1 activity was increased in the acute phase and at 3 months, D-dimer levels were significantly higher at 1 week and 1 month after stroke, whereas t-PA:Ag, TAT and F 1+2 levels remained stable during the whole study period. CONCLUSIONS: The changes of the fibrinolytic and coagulation system activity in the patients with mild or moderate ischemic stroke appeared minor compared with the results of previous studies, which included more severely ill patients. PMID- 15355489 TI - The impact of cardiac complications on outcome in the SAH population. AB - OBJECTIVES: To determine the impact of cardiac complications (CdCs) on outcomes in patients with acute subarachnoid hemorrhage (SAH). PATIENTS AND METHODS: Eighty-one adult aneurysmal SAH patients with a fisher grade >1 and/or a Hunt and Hess grade >2 were recruited for this study. CdCs were defined as electrocardiogram (ECG) changes, myocardial necrosis, arrythmias, or pulmonary edema. Outcomes were assessed at 3, 6 and 12 months by telephone interview using the Modified Rankin Scale (MRS), Glasgow Outcome Scale (GOS), Barthel Index and Medical Outcome study Short Form-36 (SF-36). RESULTS: The CdCs occurred in 33% of patients. The most common CdCs were arrythmias and pulmonary edema (30%). There was no significant difference in mortality between the two groups. At 3 months there was a significant difference in the Barthel (P = 0.007) and the SF-36 (P = 0.014) with trends in the GOS (P = 0.049) and the MRS (P = 0.063). At 6 months a significant difference remained in the SF-36 (P = 0.028) and a trend in the Barthel (P = 0.069). CONCLUSION: Results show that CdCs may negatively impact outcomes in SAH patients up to 6 months following hemorrhage. PMID- 15355490 TI - Vasoreactivity in patients with periventricular white matter lucency. AB - OBJECTIVES: Cerebral hypoperfusion has been evidenced in patients with periventricular white matter lucency (PWML), however, our knowledge is limited regarding vasoreactivity (VR) changes in these patients. Therefore, we compared the cerebral blood flow velocity (CBFV) responses during different vasoregulatory challenges in healthy volunteers, to those in patients with PWML. MATERIAL AND METHODS: In 20 patients with PWML and in 20 healthy volunteers the VR of the middle cerebral artery (MCA) system was measured by analyzing the changes of CBFV during different stimulation paradigms (ventilation, tilting and acetazolamide tests). During transcranial Doppler (TCD) registration the systemic blood pressure, the expiratory partial CO(2) pressure (pCO(2)) and the electroencephalograph (EEG) were monitored. RESULTS: The relative velocity change was significantly smaller in the PWML group than in the normal control group during hypercapnia (16 +/- 12% vs 32 +/- 17%; P < 0.001) and this finding was confirmed by assessment of pCO(2)-corrected velocity change (4.7 +/- 3.7 cm/s/kPa vs 18.4 +/- 6.8 cm/s/KPa; P < 0.001). Although CBFV measurements during acetazolamide test tended to support these findings, the changes of other parameters measured did not reach the level of significance. One patient showed considerable orthostatic reaction (mean arterial blood pressure decrease by 70 mmHg) but it was not associated with significant changes in CBFV. CONCLUSION: Patients with PWML showed an impaired VR in the MCA flow territory supporting the concept of the microangiopathic origin of leukoaraiotic changes. PMID- 15355491 TI - The effect of monoamine oxidase B (MAOB) and catechol-O-methyltransferase (COMT) polymorphisms on levodopa therapy in patients with sporadic Parkinson's disease. AB - OBJECTIVES: The etiology of sporadic idiopathic Parkinson's disease (PD) is considered multifactorial with both genetic and environmental factors modifying the disease expression. Recent studies suggest that polymorphism in monoamine oxidase B (MAOB) and catechol-O-methyltransferase (COMT) might influence the risk and treatment of PD. The aim of the study was to evaluate the effect of MAOB and COMT genetic polymorphism on effective daily dose of levodopa applied during the first 5 years of treatment, and to find out if a relationship exists between MAOB and COMT haplotypes and motor disturbances onset in PD patients treated with levodopa preparations. MATERIALS AND METHODS: A total of 95 patients (40 females and 55 males) of Polish origin diagnosed with sporadic PD were enrolled into the study, and were divided into two groups. Group 1 - patients treated with doses of levodopa below 500 mg/day during the first 5 years of treatment. Group 2 - patients requiring levodopa doses exceeding 500 mg/24 h during the first 5 years of treatment. Low activity alleles of MAOB and COMT, i.e. MAOB allele A and COMT(L) as well as high activity ones, i.e. MAOB allele G and COMT(H), were determined using PCR-RFLP method. RESULTS: No statistically significant differences were found in MAOB and COMT allele distribution in the two groups. However, the frequency of COMT(L/L) homozygotes was higher in the group treated with low doses of levodopa when compared with the second group. MAOB and COMT AG HH haplotype predominated in the group of females treated with high daily doses of levodopa when compared with AG-LL haplotype in the group of females treated with low daily doses of levodopa (<500 mg/24 h). CONCLUSION: The results of the study suggest that patients with COMT(L/L) genotype and possibly MAOB genotype A may benefit from more efficient and safer levodopa therapy. PMID- 15355492 TI - Occurrence of beta-methylamino-l-alanine (BMAA) in ALS/PDC patients from Guam. AB - We tested the brain tissues of the Chamorro people of Guam who died of amyotrophic lateral sclerosis/Parkinsonism dimentia complex (ALS/PDC) for the neurotoxin beta-methylamino-l-alanine (BMAA). We used validated high-pressure liquid chromatography and liquid chromatography-mass spectrometry analyses to test well-characterized archival tissues of the superior frontal gyrus from eight Chamorros from Guam and a comparison group of 15 Canadians. BMAA was found as a free amino acid in 83% of Chamorro ALS/PDC patients (3-10 microg/g) as a protein associated amino acid in 100% of the Chamorro individuals (149-1190 microg/g). Both forms of BMAA were also found at comparable levels in two Canadians who died of progressive neurodegenerative disease. BMAA, which is produced by cyanobacteria, may be associated with some cases of neurodegenerative disease. PMID- 15355493 TI - MG with distal muscle involvement. PMID- 15355496 TI - Clinical grade cell manipulation. AB - In the past decade, a new form of therapy based on biological rather than pharmacological intervention has been developed. The term 'cell therapy', as applied to this new therapeutic tool, means the administration of living, non germline somatic cells to humans for diagnostic or therapeutic purposes. Cell therapy products (CTPs) are generated by ex vivo processes, which comprise cell harvesting from patients or healthy donors, in vitro manipulation and administration of the manipulated cells to patients. The aim of ex vivo processes is to obtain cell subsets with defined functional properties that are capable of replacing or repairing damaged tissues or organs. Some examples of cell therapy are transplantation of expanded haematopoietic stem cells (HSCs), adoptive immunotherapy and dendritic cell vaccination to augment or restore the immune response for the treatment of malignant or infectious diseases. The types of cells most frequently used for cell therapy include haematopoietic pluripotent progenitor and stem cells from the bone marrow and peripheral blood, T-cell clones and dendritic cells. Although CTPs should be produced according to good manufacturing practice, they differ from traditional pharmaceutical products with regard to quality control and safety aspects. These differences prompted the development of a number of documents issued by regulatory bodies, which specifically address CTPs. This review discusses several issues related to the design, construction and validation of a hospital-based facility for the production of CTPs, the implementation of cell-manipulation processes and quality control of the final products. PMID- 15355497 TI - Current status and prospects for gene therapy. AB - Gene therapy is a new and exciting therapeutic concept that offers the promise of cure for an array of inherited, malignant and infectious disorders. After years of failure, substantial progress in the efficiency of gene-transfer technology has recently resulted in impressive clinical success in infants with immunodeficiency. Two of these children have, however, subsequently developed leukaemia as a result of insertional mutagenesis, raising concerns about the safety of genetic therapeutics. The purpose of this article is to review the current status of gene therapy in light of recent successes and tragedies, and to consider the challenges faced by this relatively new field. PMID- 15355498 TI - Pathogen reduction of buffy coat platelet concentrates using riboflavin and light: comparisons with pathogen-reduction technology-treated apheresis platelet products. AB - BACKGROUND AND OBJECTIVES: A pathogen-reduction technology (PRT) system using riboflavin and light has been developed for the treatment of platelet concentrates (PC) obtained by either buffy coat preparation (BCPC) or apheresis procedures (APPC). The aim of this study was to evaluate the effects of the treatment process on in vitro cell quality and on riboflavin conversion in PC. MATERIALS AND METHODS: BCPC were prepared with the Compomat G4 from whole blood which had been stored overnight after collection. APPC were obtained using the TRIMA apheresis procedure. Both PC products had been stored for 18-24 h prior to PRT treatment. BCPC and APPC were treated with PRT on day 2 and day 1 of shelf life, respectively. The treated PCs were then maintained for an additional 5 days after the PRT treatment. A panel of cell quality assays and high-performance liquid chromatography (HPLC) analysis were performed. RESULTS: Cell counts and plasma lactate dehydrogenase (LDH) levels during storage indicated that PRT did not induce significant cell lysis. Acceleration of a decrease in glucose and an increase in lactate was observed for treated PCs, but no significant differences were observed between treated BCPC and APPC. The pH of treated samples remained above 7.0, although was lower than that of the control. Platelet morphology of BCPC and APPC was well preserved. P-selectin expression indicated significant platelet activation when compared with control PC (BCPC on day 6: 39% vs. 12%; APPC on day 5: 35% vs. 18%). Both P-selectin expression and microparticle formation were not significantly different between treated BCPC and APPC during storage. The JC-1 assay also displayed no loss of mitochondria integrity during the storage of treated products. Approximately 20% of riboflavin converted into photoproducts, including lumichrome. CONCLUSIONS: PRT treatment had an effect on the development of the normal platelet storage lesion at a level which seems tolerable for clinical usage. PMID- 15355499 TI - External quality assessment for the detection of blood-borne viruses in plasma by nucleic acid amplification technology: the first human immunodeficiency virus and hepatitis B virus studies (HIV EQA/1 and HBV EQA/1) and the fifth hepatitis C virus study (HCV EQA/5). AB - BACKGROUND AND OBJECTIVES: This External Quality Assessment (EQA) study was aimed at assessing the proficiency of blood centres and blood product manufacturers in detecting, by nucleic acid amplification technology (NAT), the possible contamination of plasma with hepatitis C virus (HCV), human immunodeficiency virus (HIV) and hepatitis B virus (HBV). MATERIALS AND METHODS: Three independent panels, one for each virus, were prepared at the Istituto Superiore di Sanita (ISS) by diluting the respective reference preparations. NAT methods used by the EQA participants included polymerase chain reaction (PCR) assays by Roche, transcription-mediated amplification (TMA) assays by Chiron and in-house PCR assays. RESULTS: Forty-three of the 45 participants (95.6%) in the HCV EQA/5 who used a validated method were consistently able to detect a nominal concentration of 100 IU/ml for all six major genotypes. In the case of the HIV EQA/1, all 35 participants detected the samples containing 1000 IU/ml HIV, while five (14.3%) did not identify the samples containing 100 IU/ml HIV. With respect to the HBV EQA/1, all 16 participants correctly identified the positive samples containing either 1000 IU/ml or 100 IU/ml HBV. No false-positive results were observed with any of the three panels. CONCLUSIONS: The HCV EQA/5 showed an improved proficiency of laboratories as compared with the HCV EQA/4. In fact, HCV genotypes 1, 2, 3 and 5 were correctly identified in 100% of the assays and genotypes 4 and 6 in 97.8% of the assays. While most of the participants in the HIV EQA/1 showed a good level of proficiency, an excellent performance was shown by all participants in the HBV EQA/1. PMID- 15355500 TI - Platelet characteristic antigens of CD34+ cells in cryopreserved cord blood: a study of platelet-derived microparticles in transplant processing. AB - BACKGROUND AND OBJECTIVES: In previous studies, we found that platelet microparticles (PMPs) bind to cord blood (CB) CD34+ cells and transfer adhesion molecules to them, which enhances their engraftment. Before applying this phenomenon in actual transplants, we investigated the effect of PMPs on cryopreserved CD34+ cells in CB. MATERIALS AND METHODS: We cryopreserved 18 CB units, then evaluated the binding of PMPs to CD34+ cells after thawing, by varying the expression of platelet characteristic antigens (CD41a, CD61, CD62P and CXCR4) on these cells. Adherence of the CD34+ cells, coated with freeze/thaw induced PMPs, to endothelium and fibronectin was also studied, as were the effects of thrombin-induced PMPs from both fresh and preserved CB platelets. RESULTS: PMPs induced by freezing and thawing adhered less well to CD34+ cells than did those from fresh CB, and cells coated with these PMPs had poor adherence. However, thrombin-induced PMPs from both fresh and preserved CB platelets bound equally well to cryopreserved CD34+ cells and improved their adhesion properties. CONCLUSIONS: PMPs could be a useful tool for enhancing engraftment after CB transplantation. PMID- 15355501 TI - An update on predeposit autologous blood donation and transfusion in Europe. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to update previous data on the practice of autologous blood transfusion in Europe. MATERIALS AND METHODS: A questionnaire, mainly about predeposit, was distributed to each National Representative on the Council of Europe's Committee of Experts on Blood Transfusion and Haematology. Detailed replies were received from 23. RESULTS: Predeposit autologous blood units collected in 2000 amounted to 3.3% of the allogeneic units, compared with 4.2% in 1997. Predeposit collection was commonest in Italy (7.8%) and Germany (6.4%). CONCLUSIONS: Although autologous transfusion showed no sign of increase in Europe from 1997 to 2000, the need for alternatives to allogeneic transfusion is growing. Policy and practices in this field should be followed on an ongoing basis. PMID- 15355502 TI - LU21: a new high-frequency antigen in the Lutheran blood group system. AB - BACKGROUND AND OBJECTIVES: The Lutheran blood group system comprises 18 antigens numbered LU1 to LU20, with two numbers obsolete. Thirteen antigens are of high frequency. MATERIALS AND METHODS: Serological tests were performed by conventional methods. The monoclonal antibody-specific immobilization of erythrocyte antigens (MAIEA) assay was carried out with monoclonal antibodies to Lutheran glycoprotein. All exons of the LU gene were amplified by the polymerase chain reaction (PCR) and directly sequenced from genomic DNA. RESULTS: A patient was found to have an antibody to a high-frequency red cell antigen during her second pregnancy. The antibody was shown to be Lutheran-related and was distinguished from all reported Lutheran antibodies. MAIEA suggested the antibody was defining a novel epitope in domain 1 of the Lu-glycoprotein. Sequencing of the LU gene revealed a new homozygous single-point mutation, C282G, in exon 3, encoding an Asp94Glu change in the first domain of the Lu-glycoprotein. CONCLUSIONS: The antibody detected a new high-frequency Lutheran antigen, numbered LU21, that appears to result from an Asp94Glu substitution in the first, N-terminal domain of the Lu-glycoprotein. PMID- 15355503 TI - High-throughput genotyping of human platelet antigens using the 5'-nuclease assay and minor groove binder probe technology. AB - BACKGROUND AND OBJECTIVES: Human platelet antigen (HPA) genotyping is a valuable tool for typing platelets to assist in the management of alloimmunized thrombocytopenic patients. We describe, for the first time, 5' nuclease assays (NA) to genotype HPA-5 and -15, and improved 5'-NA to genotype HPA-1, -2 and -3, by utilizing minor groove binder (MGB) and non-fluorescent quencher (NFQ) technology. Superior probe specificity and fluorescent performance is attained through MGB-NFQ probe modifications compared with previous 5'-NA designs. MATERIALS AND METHODS: Primers and dye-labelled MGB-NFQ probes were designed and synthesized to detect the single nucleotide polymorphism responsible for each HPA 1, -2, -3, -5 and -15. One-hundred blood samples were tested for the combinations of HPA genotypes 1, 2, 3 and 5 by our traditional sequence-specific primer polymerase chain reaction (SSP-PCR) method, and 41 blood samples were tested for HPA-15 by SSP-PCR at an external laboratory. These results were then compared with those obtained by using the new 5'-NA. RESULTS: There was complete concordance of results for all samples tested by SSP-PCR and 5'-NA. The 5'-NA offers distinct advantages over non-fluorescent genotyping methods. DNA amplification and allele discrimination occurs in a single closed tube for each antigen with no post-PCR manipulation required. This minimizes the risk of cross contamination and mislabelling of samples, as well as making the assay less time consuming to perform. In comparison with other fluorescent assays, the 5'-NA has the highest sample throughput, resulting from the use of a 96-well platform, identical cycling conditions for all assays and the potential for automation. CONCLUSIONS: Genotyping for HPA-1, -2, -3, -5, and -15 by the 5'-NA is suitable for routine analysis. The latest 5'-NA design, using MGB probe technology, ensures superior detection of all alleles and is the most versatile fluorescent assay, ideal for both urgent clinical samples and large-scale screening programs. PMID- 15355504 TI - Polymorphism of the human platelet antigen-5 system is a risk factor for occlusive vascular complications in patients with sickle cell anemia. AB - BACKGROUND: Polymorphisms of platelet membrane glycoproteins such as human platelet antigen (HPA)-1b, HPA-2b, the -5T/C Kozak sequence and C807T have been described as risk factors for vascular disease. Vaso-occlusion episodes are a common feature of sickle cell anaemia (SCA), leading to complications such as stroke, acute chest syndrome, avascular head femur necrosis and priapism. Complex interactions are involved in vaso-occlusion, and activated platelets may play an important role. These data raised the question of whether platelet polymorphisms could be implicated in occlusive vascular complications (OVC) of SCA. MATERIALS AND METHODS: In this study, 97 patients with SCA were analysed in two groups: 34 patients presenting with OVC (SCA-VC) and 63 without these complications (SCA-N). The distribution of the HPA-1, -2 and -5 systems, as well as C807T dimorphism and -5T/C Kozak sequence alleles, was evaluated using DNA-based methods. RESULTS: Patients of the SCA-VC group showed a higher frequency of the HPA-5b allele (0.324) compared with those of the SCA-N group (0.111) (chi2 = 13.19, P = 0.0002). None of the other polymorphisms, isolated or associated as haplotypes, demonstrated any correlation with the development of OVC in these patients. CONCLUSIONS: The findings of this study suggest that the HPA-5b allele is a genetic risk factor for the development of OVC in patients with SCA. This allele could be explored as a target for the development of new therapeutic approaches. PMID- 15355506 TI - Are current measures to prevent transfusion-associated protozoal infections sufficient? PMID- 15355505 TI - Transfusion-transmitted bacterial infection: risks, sources and interventions. PMID- 15355508 TI - Retraction: Finfer S, Norton R, Bellomo R, Boyce N, French J, Myburch J, on behalf of the SAFE Study Investigators. The SAFE study: saline vs. albumin for fluid resuscitation in the critically ill. Vox Sang 2004; 87 (Suppl. 2): S123 S131. PMID- 15355509 TI - Endocytic and transcytotic processes in villous syncytiotrophoblast: role in nutrient transport to the human fetus. AB - The supply of nutrients to the developing fetus is a major function of the human hemochorial placenta, a placenta type in which the fetal chorion is in direct contact with the maternal blood. At term, nutrients have to be transported across two cell layers in chorionic villi, the syncytiotrophoblast (STB) and fetal endothelial cells. The STB is a continuous syncytium covering the entire surface of chorionic villi. This polarized epithelium is specialized in exchange processes and membrane trafficking between the apical membrane facing the maternal blood and the basal membrane facing the fetal endothelium. To meet placental and fetal requirements, the STB selectively takes up and transports a variety of nutrients, hormones, growth factors and cytokines and also transfers passive immunity to the fetus by receptor-mediated transcytosis. In this review in vivo and in vitro systems currently used to study STB functions are discussed and the potential mechanisms of transplacental IgG, iron, lipoprotein and glucose transport are presented. As revealed in this article, the placenta is a tissue where intensive cell biological research is required to unravel endocytic trafficking pathways in a highly specialized cell such as the STB. PMID- 15355510 TI - Insulin and oleate promote translocation of inosine-5' monophosphate dehydrogenase to lipid bodies. AB - In the present study we identify inosine-5' monophosphate dehydrogenase (IMPDH), a key enzyme in de novo guanine nucleotide biosynthesis, as a novel lipid body associated protein. To identify new targets of insulin we performed a comprehensive 2-DE analysis of (32)P-labelled proteins isolated from 3T3-L1 adipocytes (Hill et al. J Biol Chem 2000; 275: 24313-24320). IMPDH was identified by liquid chromatography/tandem mass spectrometry as a protein which was phosphorylated in a phosphatidylinositol (PI) 3-kinase-dependent manner upon insulin treatment. Although insulin had no significant effect on IMPDH activity, we observed translocation of IMPDH to lipid bodies following insulin treatment. Induction of lipid body formation with oleic acid promoted dramatic redistribution of IMPDH to lipid bodies, which appeared to be in contact with the endoplasmic reticulum, the site of lipid body synthesis and recycling. Inhibition of PI 3-kinase blocked insulin- and oleate-induced translocation of IMPDH and reduced oleate-induced lipid accumulation. However, we found no evidence of oleate-induced IMPDH phosphorylation, suggesting phosphorylation and translocation may not be coupled events. These data support a role for IMPDH in the dynamic regulation of lipid bodies and fatty acid metabolism and regulation of its activity by subcellular redistribution in response to extracellular factors that modify lipid metabolism. PMID- 15355511 TI - Temperature-sensitive random insulin granule diffusion is a prerequisite for recruiting granules for release. AB - Glucose-evoked insulin secretion exhibits a biphasic time course and is associated with accelerated intracellular granule movement. We combined live confocal imaging of EGFP-labelled insulin granules with capacitance measurements of exocytosis in clonal INS-1 cells to explore the relation between distinct random and directed modes of insulin granule movement, as well as exocytotic capacity. Reducing the temperature from 34 degrees C to 24 degrees C caused a dramatic 81% drop in the frequency of directed events, but reduced directed velocities by a mere 25%. The much stronger temperature sensitivity of the frequency of directed events (estimated energy of activation approximately 135 kJ/mol) than that of the granule velocities (approximately 22 kJ/mol) suggests that cooling-induced suppression of insulin granule movement is attributable to factors other than reduced motor protein adenosine 5'-triphosphatase activity. Indeed, cooling suppresses random granule diffusion by approximately 50%. In the single cell, the number of directed events depends on the extent of granule diffusion. Finally, single-cell exocytosis exhibits a biphasic pattern corresponding to that observed in vivo, and only the component reflecting 2nd phase insulin secretion is affected by cooling. We conclude that random diffusive movement is a prerequisite for directed insulin granule transport and for the recruitment of insulin granules released during 2nd phase insulin secretion. PMID- 15355512 TI - Insulin stimulates the entry of GLUT4 into the endosomal recycling pathway by a quantal mechanism. AB - The insulin-sensitive glucose transporter GLUT4 mediates the uptake of glucose into adipocytes and muscle cells. In this study we have used a novel 96-well plate fluorescence assay to study the kinetics of GLUT4 trafficking in 3T3-L1 adipocytes. We have found evidence for a graded release mechanism whereby GLUT4 is released into the plasma membrane recycling system in a nonkinetic manner as follows: the kinetics of appearance of GLUT4 at the plasma membrane is independent of the insulin concentration; a large proportion of GLUT4 molecules do not participate in plasma membrane recycling in the absence of insulin; and with increasing insulin there is an incremental increase in the total number of GLUT4 molecules participating in the recycling pathway rather than simply an increased rate of recycling. We propose a model whereby GLUT4 is stored in a compartment that is disengaged from the plasma membrane recycling system in the basal state. In response to insulin, GLUT4 is quantally released from this compartment in a pulsatile manner, leaving some sequestered from the recycling pathway even in conditions of excess insulin. Once disengaged from this location we suggest that in the continuous presence of insulin this quanta of GLUT4 continuously recycles to the plasma membrane, possibly via non-endosomal carriers that are formed at the perinuclear region. PMID- 15355513 TI - Missorting of LaCrosse virus nucleocapsid protein by the interferon-induced MxA GTPase involves smooth ER membranes. AB - The interferon-induced human MxA protein belongs to the class of dynamin-like, large guanosine-5'-triphosphatases that are involved in intracellular vesicle trafficking and organelle homeostasis. MxA shares many properties with the other members of this protein superfamily, including the propensity to self-assemble and to associate with lipid membranes. However, MxA is unique in that it has antiviral activity and inhibits the replication of several RNA viruses. Here, we determined the role of membranes for the antiviral function of MxA using LaCrosse bunyavirus (LACV). We show that MxA does not affect trafficking and sorting of viral glycoproteins but binds and mislocates the viral nucleocapsid (N) protein into membrane-associated, large perinuclear complexes. We further demonstrate that MxA localizes to a subcompartment of the smooth endoplasmic reticulum where the viral N protein accumulates. In infected MxA-expressing cells, oligomeric MxA/N complexes are formed in close association with COP-I-positive vesicular tubular membranes. Our results suggest that this membrane compartment is the preferred place where MxA and N interact, leading to efficient sequestration and missorting of an essential viral component. PMID- 15355514 TI - Rab coupling protein associates with phagosomes and regulates recycling from the phagosomal compartment. AB - The Rab coupling protein (RCP) is a recently identified novel protein that belongs to the Rab11-FIP family. RCP interacts specifically with Rab4 and Rab11, small guanosine-5'-triphosphatases that function as regulators along the endosomal recycling pathway. We used fluorescence confocal microscopy and biochemical approaches to evaluate the participation of RCP during particle uptake and phagosome maturation. In macrophages, RCP is predominantly membrane bound and displays a punctuate vesicular pattern throughout the cytoplasm. RCP is mainly associated with transferrin-containing structures and Rab11-labeled endosomes. Overexpression of H13, the carboxyl-terminal region of RCP that contains the Rab binding domain, results in an abnormal endosomal compartment. Interestingly, we found that RCP is associated as discrete patches or protein domains to early phagosomal membranes. In macrophages, overexpression of full length RCP stimulates recycling from the phagosomal compartment, whereas overexpression of H13 diminishes this vesicular transport step. It is likely that acting as an intermediate between Rab4 and Rab11, RCP regulates membrane flux along the phagocytic pathway via recycling events. PMID- 15355515 TI - Regulation of myosin-VI targeting to endocytic compartments. AB - Myosin-VI has been implicated in endocytic trafficking at both the clathrin coated and uncoated vesicle stages. The identification of alternative splice forms led to the suggestion that splicing defines the vesicle type to which myosin-VI is recruited. In contrast to this hypothesis, we find that in all cell types examined, myosin-VI is associated with uncoated endocytic vesicles, regardless of splice form. GIPC, a PDZ-domain containing adapter protein, co assembles with myosin-VI on these vesicles. Myosin-VI is only recruited to clathrin-coated vesicles in cells that express high levels of Dab2, a clathrin binding adapter protein. Overexpression of Dab2 is sufficient to reroute myosin VI to clathrin-coated pits in cells where myosin-VI is normally associated with uncoated vesicles. In normal rat kidney (NRK) cells, which express high endogenous levels of Dab2, splicing of the globular tail domain further modulates targeting of ectopically expressed myosin-VI. Although myosin-VI can be recruited to clathrin-coated pits, we find no requirement for myosin-VI motor activity in endocytosis in NRK cells. Instead, our data suggest that myosin-VI recruitment to clathrin-coated pits may be an early step in the recruitment of GIPC to the vesicle surface. PMID- 15355517 TI - Dimensions of informal care and quality of life among elderly family caregivers. AB - The aim was to investigate dimensions of caregiving activities among elderly (75+) caregivers based on Nolan's model and to study the dimensions in relation to health-related quality of life (Short Form 12). Responses to a Swedish postal survey (n=4278, response rate 75-79 years old: 60%; 80-84: 56%; 85-89: 48% and 90+: 42%) showed that 783 persons (18%) were helping another person due to that person's impaired health, 41.6% women, mean age for women 81.8 years (SD 4.96) and for men 81.7 years (SD 4.32). The postal questionnaire included SF-12, demographic data and questions about caregiving activities derived from Nolan's model, social network and contacts with health care. Adapting their activities to be prepared if something happened (52%), having regular contact to prevent problems (35%), helping in contacts with the hospital (57%), helping with instrumental activities of daily living (49%), personal activities of daily living (14%), medical care (11%) and helping to improve functions (14%) were the activities reported. Adapting own activities, regular contact, weak economy and needing instrumental help with daily living oneself predicted low MCS12. The importance of early involvement on the part of the caregivers was emphasized. PMID- 15355518 TI - The application of grounded theory and symbolic interactionism. AB - This paper describes the methodological and theoretical context and underpinnings of a study that examined community psychiatric nurses' work with family caregivers of older people with depression. The study used grounded theory research methods, with its theoretical foundations drawn from symbolic interactionism. The aims of the study were to describe and conceptualize the processes involved when community nurses work and interact with family caregivers and to develop an explanatory theory of these processes. This paper begins with an explanation of the rationale for using grounded theory as the method of choice, followed by a discussion of the theoretical underpinnings of the study, including a brief summary of the nature and origins of symbolic interactionism. Key premises of symbolic interactionism regarded as central to the study are outlined and an analytical overview of the grounded theory method is provided. The paper concludes with a commentary on some of the issues and debates in the use of grounded theory in nursing research. The main purpose of this paper is to provide a methodical and critical review of symbolic interactionism and grounded theory that can help readers, particularly those who are intending to use grounded theory, better understand the processes involved in applying this method to their research. PMID- 15355519 TI - Nurses' personal statements about factors that influence their decisions about the time they spend with residents with long-term mental illness living in psychiatric group dwellings. AB - One seldom-discussed issue is the factors that influence nurses' decisions about the time they spend with residents in psychiatric care. This study uses a qualitative naturalistic approach and consists of an analysis of focus-group interviews with nurses, which aimed to identify factors affecting nurses' decisions about being with or being nonattendant in their relationship with their residents. Two series of focus-group interviews were conducted, interpreted and analysed through content analysis. The study included all the staff (n=32) at two municipal psychiatric group dwellings housing residents mainly with a diagnosis of long-term schizophrenia. This study revealed that the main factor that determined nurses' nurse/resident time together or nonattendance time was whether they liked or disliked the individual resident. One possible explanation is the carers' change from a perspective in which the nursing care was given on the basis of each resident's needs and rights, based on the individual nurse's professional judgement, to a consumer perspective, which leads to a change in responsibility from themselves to the individual residents. PMID- 15355520 TI - The progression of suffering implies alleviated suffering. AB - There is a lack of research focusing directly on both patients' suffering and alleviated suffering in relation to care. The aim of this paper was to investigate the progression of suffering in relation to the encounter between the suffering person and the caregiver from the perspective of an understanding of life. The progression of suffering is assumed to be an existential 'sign' of the development of understanding of life as an ontological or spiritual entity, which demands a meaning-creating encounter between the patient and caregiver. The concept 'existential caring encounter' was used to describe how the encounter between patient and caregiver can create meaning in communion and thereby alleviate suffering by making it bearable. The study was carried out using an interpretive, hermeneutic approach. The study as a whole comprises three parts, and these include letter-writing and interviews. The findings are described by the following main theses: (a) a darkness in life understanding is existentially experienced as unbearable suffering and requires an encounter involving attentive care and confrontation; (b) the turning point means that the struggle of suffering begins; and (c) the encounter involves being meaning-creating in a communion in the struggle of suffering. An understanding of the patterns of unbearable and bearable suffering can be of help to the caregiver in caring for the patient by serving as a basis for meaning-creation in communion. This may thereby be a way of alleviating the patient's suffering by making it bearable during the progression of suffering. PMID- 15355521 TI - Patients' voices on satisfaction: unheeded women and maltreated men? AB - To capture the differences in female and male patients' hospital experiences, this article discusses the gendered nature of 699 free-text comments given in a survey of patient satisfaction at a Norwegian university hospital. The comments were analysed both quantitatively and qualitatively, and the impressions gained through the comments were then to a smaller extent compared with the patients' multiple choice questionnaire ticking. Female responders added free-text comments to their multiple-choice responses more often than male responders did. Male and female patients also drew attention to different aspects of what it was like to be hospitalized, and male and female patients also expressed themselves differently. The voices of the patients were mainly the voices of contextualizing experiences, reflecting differences in male and female patient's experiences and in the meaning the patients attached to their experiences. The rather clear gender differences in our data emphasizes the need for a gender-sensitive approach in the encounters between staff and patients as well as in the design of patient satisfaction research. PMID- 15355522 TI - Prevalence of patients subjected to constraint in Norwegian nursing homes. AB - BACKGROUND: A recent questionnaire showed that different kinds of constraint such as physical restraint, electronic surveillance, use of force or pressure in medical treatment and in activities of daily living (ADL) are frequently used in Norwegian nursing homes. The study did not include information at patient level, and except for studies about physical restraints, we have not found any studies reporting the prevalence of various forms of constraint. AIM: To describe the prevalence of various types of constraint in Norwegian nursing homes. MATERIALS AND METHOD: A structured interview was carried out with the primary carers of a random sample of 1501 patients from 222 nursing-home wards in 54 municipalities representing all five health regions in Norway. Data were collected from regular units (RUs) and special care units (SCUs) for persons with dementia. Episodes of constraint during 1 week were recorded. Five main groups of constraint were aggregated, mechanical restraint, nonmechanical restraint, electronic surveillance, force or pressure in medical examination or treatment and force or pressure in ADL. RESULTS: Patients (36.7%) in RUs and 45.0% of the patients in SCUs were subjected to any constraint. Most frequent was use of mechanical restraint (23.3% in RUs; 12.8% in SCUs) and use of force or pressure in ADL (20.9% in SCUs; 16.6% in RUs). Use of force or pressure in medical examination or treatment was more frequent used in SCUs (19.1%) compared with RUs (13.5%). Nonmechanical restraint was less frequently used (8.3% in SCUs; 3.0% in RUs) and electronic surveillance was seldom used (7.2% in RUs; 0.9% in SCUs). CONCLUSION: The use of constraint is a problem in Norwegian nursing homes. Studies are needed to learn more about why constraint is used, and if there is patient or ward characteristics that can explain the use of constraint. PMID- 15355523 TI - Midwives' attitudes to and knowledge about a newly introduced foetal screening method. AB - A number of screening procedures are offered by midwives during pregnancy and the number is increasing rapidly. The measurement of nuchal translucency is a fairly new ultrasound method for antenatal screening, primarily for Down syndrome. The results give a better risk calculation than maternal age alone and can mean a decrease in the number of invasive procedures needed to identify this syndrome prenatally. The aim of this study was to gain insight into the midwives' point of view concerning the introduction of the method in two different hospitals. In one hospital it had been introduced as part of a research project while in the other it had been integrated as an offer in the antenatal care programme. A questionnaire was sent to the 80 midwives working in the antenatal clinics serving these two hospitals. A total of 79% of the questionnaires were answered. The results indicate that in both districts, the similarities are greater than the differences with regard to the midwives' education, knowledge and their own opinions of their ability to inform pregnant women about the method. Although most of the respondents were positive to it, a number of midwives felt that, in general, information about foetal diagnosis was a difficult part of their work, mentioning both ethical and practical aspects. This highlights the need for continuing education, standardized policy and an ongoing ethical debate. PMID- 15355524 TI - Migraine and chronic daily headache management: a qualitative study of patients' perceptions. AB - AIM: The aim of the study was to gain insight into the patients' perceptions of migraine and chronic daily headache (CDH) management. METHODS: Thirteen, semi structured and individual interviews with seven migraine and five CDH patients were carried out and analysed in QSR NUD*IST5, using a grounded theory methodology. RESULTS: The participants described using five areas of management: 1) health care use; 2) medication use; 3) alternative therapies; 4) social support; and 5) lifestyle and self-help. The participants described their expectations, preferences, worries and (dis)satisfaction in relation to these five areas of management. The participants adapted headache management to suit their needs and preferences, making migraine and CDH management highly individual and giving the headache patient a central role within their own care. CONCLUSION: Health care is changing towards a greater involvement of the patients in their own care. Therefore, it is important to increase understanding of the patients' perspective of chronic diseases, including migraine and CDH. The results from this study inform health care professionals of the range of their patients' needs and preferences. This knowledge can be used to shape clinical practice, to develop patient education programmes and to further research efforts into issues that are important to the headache patient. PMID- 15355525 TI - The Swedish version of the PACU-Behavioural Pain Rating Scale: a reliable method of assessing postoperative pain? AB - One of the obstacles to effective pain management in the Post-Anaesthesia Care Unit (PACU) is the lack of systematic and comprehensive methods for assessing and treating postoperative pain. Nurses' intuitive knowledge of pain were verbalized and divided into four categories of pain behaviours in the PACU-Behavioural Pain Rating Scale. The aim of the present study was to test the reliability of the Swedish version by performing test-retest and interrater reliability in clinical conditions. Materials for this study were gathered through patient observations. The results showed poor agreement in the item restless, fair agreements in both items tense muscles and frowning and grimacing, while patients' sounds showed moderate agreement. Interrater reliability showed moderate agreement in the item restless, very good agreement in tense muscles and patient sounds, and good agreement in frowning and grimacing. A refined classification of the separate categories and a careful review of the category of behaviours is suggested. A more detailed manual for the categories needs to be developed to establish more acceptable reliability. PMID- 15355526 TI - Electronic nursing documentation in primary health care. AB - The aim of this study was to describe and analyse nursing documentation based on an electronic patient record (EPR) system in primary health care (PHC) with emphasis on the nurses' opinions and what, according to the nursing process and the use of the keywords, the nurses documented. The study was performed in one county council in the south of Sweden and included 42 Primary Health Care Centres (PHCC). It consisted of a survey, an audit of nursing records with the Cat-ch-Ing instrument and calculation of frequencies of keywords used during a 1-year period. For the survey, district nurses received a postal questionnaire. The results from the survey indicated an overall positive tendency concerning the district nurses' opinions on documentation. Lack of in-service training in nursing documentation was noted and requested from the district nurses. All three parts of the study showed that the keywords nursing interventions and status were frequently used while nursing diagnosis and goal were infrequent. From the audit, it was noted that medical status and interventions appeared more often than nursing status. The study demonstrated limitations in the nursing documentation that inhibited the possibility of using it to evaluate the care given. In order to develop the nursing documentation, there is a need for support and education to strengthen the district nurses' professional identity. Involvement from the heads of the PHCC and the manufactures of the EPR system is necessary, in cooperation with the district nurses, to render the nursing documentation suitable for future use in the evaluation and development of care. PMID- 15355527 TI - Testing of an audit instrument for the nursing discharge note in the patient record. AB - OBJECTIVES: This study validates and tests the reliability of an audit instrument constructed to evaluate the content of nursing discharge notes. DESIGN: Instrument validation and reliability testing. MAIN OUTCOME MEASURES: Factor analysis identifying structure through data summarization of the instrument, association between scores in test-retest, and interrater reliability between auditors. RESULTS: VALIDITY: Three factors emerged in the factor analysis: 'General information', 'Planning', and 'Assessment', accounting for 76% of the variance regarding the quantitative aspect and 79% of the variance regarding the qualitative aspect, confirming the distinctiveness. Reliability: The Spearman rank-order correlation coefficient calculated per item in the test-retest ranged from 0.72 to 1.0 (p=0.01). The correlation coefficient for the total score was 0.98 (p=0.01). There were no differences in item scores between the test and retest in 93% of the comparisons (n=486). Between the two auditors, the Spearman rank-order correlation coefficient in each item ranged from 0.83 to 1.00 (p=0.01) and weighted kappa values from 0.70 to 1.00 with the exception of one item in both calculations. The correlation coefficient for the auditors' total score was 0.99 (p=0.01). The Student's paired t-test comparing the two auditors' mean values in five different parts of the instrument showed no significant differences in score. CONCLUSION: The Cat-ch-Ing EPI instrument shows a high reliability and validity as an audit instrument to evaluate the content of nursing discharge notes. PMID- 15355528 TI - The effect of a VIPS implementation programme on nurses' knowledge and attitudes towards documentation. AB - The Copenhagen University Hospital decided to adhere to the standards of the Joint Commission of International Accreditation in 2000. These standards require systematic assessment of patient care needs and include the use of written nursing care plans. In order to meet these standards, the hospital management decided to introduce the Swedish VIPS model, which is a model designed to structure nursing documentation (VIPS is an acronym for well-being, integrity, prevention and safety). The present study explores the nurses' knowledge and attitudes towards documentation and addresses the research questions: (a) what are the nurses' attitudes towards documentation of nursing care? and (b) do nurses have sufficient knowledge of the documentation system to systematically document their patient assessment and clinical decisions? The research design was prospective, comparative, and quasi-experimental (nonrandomized), including a study group (n=72) and a control group (n=57). A questionnaire was used to compare nurses' self-evaluated attitudes towards documentation, and a multiple choice test was given in order to assess nurses' knowledge of the documentation system. The study group participated in a special implementation programme (response rate 82%), while the control group attended the regular 3-day documentation course at the hospital (response rate 79%). The study showed that the two groups responded similarly, but the nurses in the study group were significantly stronger in their conviction that they had the knowledge to make care plans and that they routinely made them. The study group demonstrated slightly less motivation than the control group, while the two groups shared a positive attitude towards nursing documentation. The study group did consistently better on the knowledge tests. The findings show that the implementation programme had a positive impact on nursing documentation, and that the VIPS model increased the nurses' understanding of the nursing process. PMID- 15355529 TI - Ethical issues in participatory action research. AB - The purpose of this article is to describe the ethical issues arising out of participatory action research (PAR), on the basis of both an empirical study and the research literature, and to discuss how to deal with these issues. The data consist of the experiences and results of three phases of PAR relating to orthopaedic patients with rheumatoid arthritis (RA) and the analysis of 20 articles on the ethics of action research. As a result, the following ethical issues and the ways to treat them were discussed: informed consent, confidentiality and anonymity, protecting an individual from harm, the role of the researcher, the location of 'power' in PAR, and the ownership of the research. The flexibility of PAR in use and its main features are also related to the decisions made and actions taken in response to ethical issues. It is particularly important in PAR to proceed according to the participants, and to involve them from the beginning of the process, in order to insure the equal balance of power between participants and researcher. PMID- 15355530 TI - Does badger culling make TB worse? PMID- 15355531 TI - Purification and characterization of an endopeptidase that has an important role in the carboxyl terminal processing of antihypertensive peptides in Lactobacillus helveticus CM4. AB - AIMS: To purify and characterize a peptidase that can catalyse C-terminal processing of antihypertensive peptide from Lactobacillus helveticus CM4. METHODS AND RESULTS: An endopeptidase which seems to process the carboxyl terminal end of two antihypertensive peptides, Val-Pro-Pro and Ile-Pro-Pro, was purified from Lactobacillus helveticus CM4 by four stages of column chromatography, using synthetic pro-peptide as a substrate. The molecular weight of the purified enzyme was estimated to be 67,000 by SEPHACRYL S-200 and 70,000 by SDS-PAGE analysis. The purified enzyme generated: (i) Val-Pro-Pro from Val-Pro-Pro-Phe-Leu and Val Pro-Pro-Phe-Leu-Gln-Pro, and (ii) Ile-Pro-Pro from Ile-Pro-Pro-Leu-Thr and Ile Pro-Pro-Leu-Thr-Gln-Thr, but theses peptides could not be generated from Val-Pro Pro-Phe, Val-Pro-Pro-Phe-Leu-Gln, Ile-Pro-Pro-Leu and Ile-Pro-Pro-Leu-Thr-Gln. Part of the amino terminal sequence of the purified enzyme had homology to a previously reported pepO gene product. CONCLUSION: These results suggest that the purified endopeptidase isolated in this study have an important role in the carboxyl terminal processing of two antihypertensive peptides in Lact. helveticus CM4. PMID- 15355532 TI - Antibacterial effect of crude water-soluble arrowroot (Puerariae radix) tea extracts on food-borne pathogens in liquid medium. AB - AIMS: To evaluate the effect of crude water-soluble arrowroot tea extracts on microbial growth of food-borne pathogens in liquid medium and to confirm the damage to bacterial cells using Transmission Electronic Microscopy (TEM). METHODS AND RESULTS: Inhibition of growth of Escherichia coli O157:H7, Salmonella enterica serovar Enteritidis, Listeria monocytogenes and Staphylococcus aureus was investigated using Brain Heart Infusion (BHI) broth containing 0 (control), 0.63, 1.25, 2.5 and 5.0% (w/v) arrowroot tea. Bacterial cell counts were performed on specific selective agar on days 0, 1, 3 and 5. BHI containing 5.0% arrowroot tea extract showed a 6-7 log suppression of growth for all test strains on days 3 and 5, compared with the control. Even 0.63% arrowroot tea effectively inhibited microbial growth of all test strains on day 5. TEM images of the samples treated with 5.0% arrowroot tea revealed the rupture of cell walls and nonhomogeneous disposition of cytoplasmic materials within treated bacteria. CONCLUSIONS: Crude water-soluble arrowroot tea extract strongly inhibited microbial growth of all test pathogens in liquid medium. SIGNIFICANCE AND IMPACT OF THE STUDY: Water-soluble arrowroot tea extract has the potential to be used directly on foods or as a spray on the surfaces of food handling and processing facilities in order to prevent microbial growth of both Gram-negative and Gram positive bacteria. PMID- 15355533 TI - Survey of mycoflora and ochratoxin A in dried vine fruits from Argentina markets. AB - AIMS: The aims of this work were to identify the mycoflora and to evaluate the natural occurrence of OA in dried vine fruits. Likewise, the capacity to produce OA by Aspergillus section Nigri was studied. MATERIALS AND METHODS: Fifty samples of dried vine fruits were obtained from Mendoza and San Juan provinces. The surface disinfection method was used for mycoflora determination using the medium dichloran 18% glycerol agar (DG18) and dichloran Rose Bengal chloramphenicol agar (DRBC). RESULTS: Statistical analysis demonstrated that the species A. niger var. niger and Aspergillus niger var. awamori were isolated in higher frequency from black dried vine fruits from DRBC and DG18 media (P < 0.01). OA was found in 74% of the dried vine fruits samples. Sixty-two strains (28%) of Aspergillus section Nigri, were OA producers. In the species A. carbonarius the highest percentages of ochratoxigenic strains were detected (82.6%). CONCLUSIONS: The presence of ochratoxigenic strains of Nigri section in dried vine fruits suggests that they may be an important source of OA in this substrate. Dried vine fruits can also be an important source of OA people who consume large amounts. SIGNIFICANCE AND IMPACT OF THE STUDY: The dried vine fruits contamination with Aspergillus section Nigri and OA was significant. PMID- 15355534 TI - Isolating Mortierella alpina strains of high yield of arachidonic acid. AB - AIMS: To develop a fast isolation method for arachidonic acid-producing fungi of high yield. METHODS AND RESULTS: Relation between the staining degree of mycelia of Mortierella alpina stained with triphenyltetrazolium chloride (TTC) and arachidonic acid content in the fungal lipids was investigated. Results showed that staining degree of mycelia stained with TTC increased when arachidonic acid content in mycelia lipids increased. This finding was used to isolate strains of high arachidonic acid yield. Arachidonic acid producing fungi were selectively isolated from soil at a low temperature of 4 degrees C and the mycelia of these isolates were stained with TTC. CONCLUSIONS: The strain M. alpina M6 that had the highest staining degree had the highest arachidonic acid content (72.3%). The yield of arachidonic acid in this strain reached 4.82 g l(-1). SIGNIFICANCE AND IMPACT OF THE STUDY: A fast and effective method to isolate strains of high arachidonic acid yield was established according to the finding that staining degree of mycelia of M. alpina was positively correlated with arachidonic acid content in mycelia lipid. PMID- 15355535 TI - Malt sprout extract medium for cultivation of Lactobacillus plantarum protective cultures. AB - AIMS: The aim was to develop a cheap cereal-based alternative medium for the large-scale production of biopreservative Lactobacillus plantarum VTT E-79098. We examined the effect of growth medium and pH control on the cell yield of Lact. plantarum E-79098 and the antimicrobial activity of the cell-free extracts. METHODS: Fermentations using a novel Malt Sprout Extract Medium (MSE) were performed with different pH regimes. The antimicrobial activity of the cell-free extracts against Pantoea agglomerans VTT E-90396 and Fusarium avenaceum VTT D 80147 was assessed with automated turbidometry. SIGNIFICANCE AND IMPACT OF THE STUDY: When compared with MRS, the MSE medium cultures produced equal growth yields of Lact. plantarum VTT E-79098 and enhanced antimicrobial potential against the Gram-negative bacterium P. agglomerans and a Fusarium fungus. The MSE medium can be used as a low-cost alternative to MRS for producing high cell yields and good antimicrobial activity of Lact. plantarum. PMID- 15355536 TI - Characterization of Streptococcus bovis from the rumen of the dromedary camel and Rusa deer. AB - AIMS: Isolation and characterization of Streptococcus bovis from the dromedary camel and Rusa deer. METHODS AND RESULTS: Bacteria were isolated from the rumen contents of four camels and two deer fed lucerne hay by culturing on the semi selective medium MRS agar. Based on Gram morphology and RFLP analysis seven isolates, MPR1, MPR2, MPR3, MPR4, MPR5, RD09 and RD11 were selected and putatively identified as Streptococcus. The identity of these isolates was later confirmed by comparative DNA sequence analysis of the 16S rRNA gene with the homologous sequence from S. bovis strains, JB1, C14b1, NCFB2476, SbR1, SbR7 and Sb5, from cattle and sheep, and the Streptococcus equinus strain NCD01037T. The percentage similarity amongst all strains was >99%, confirming the identification of the camel isolates as S. bovis. The strains were further characterized by their ability to utilize a range of carbohydrates, the production of volatile fatty acids (VFA) and lactate and the determination of the doubling time in basal medium 10 supplemented with glucose. All the isolates produced l-lactate as a major fermentation end product, while four of five camel isolates produced VFA. The range of carbohydrates utilized by all the strains tested, including those from cattle and sheep were identical, except that all camel isolates and the deer isolate RD11 were additionally able to utilize arabinose. CONCLUSIONS: Streptococcus bovis was successfully isolated from the rumen of camels and deer, and shown by molecular and biochemical characterization to be almost identical to S. bovis isolates from cattle and sheep. SIGNIFICANCE AND IMPACT OF THE STUDY: Streptococcus bovis is considered a key lactic acid producing bacterium from the gastrointestinal tract of ruminants, and has been implicated as a causative agent of lactic acidosis. This study is the first report of the isolation and characterization of S. bovis from the dromedary camel and Rusa deer, and suggests a major contributive role of this bacterium to fermentative acidosis. PMID- 15355537 TI - Genotyping and genetic diversity of Arcobacter butzleri by amplified fragment length polymorphism (AFLP) analysis. AB - AIMS: To investigate the potential of amplified fragment length polymorphism (AFLP) profiling for genotyping Arcobacter butzleri and to obtain further data on the genetic diversity of this organism. METHODS AND RESULTS: Seventy-three isolates of Danish, British, Turkish, Swedish, Nigerian and North American origin from human infections, chickens, turkeys, ducks, sheep and poultry abbatoir effluent were studied by use of a protocol that involved stringent PCR amplification of fragments derived from digestion of genomic DNA with restriction enzymes BglII and Csp6I. The mean similarity value of duplicate profiles of 10 isolates was 91.15%, indicating the method to be reproducible. Numerical analysis of all 73 isolates distinguished 51 subtypes at the 91% similarity level, of which 39 comprised single strains. The remaining 34 isolates were distributed among 12 subtypes, each of which contained strains homogeneous with respect to their respective source of isolation. However, contemporaneous strains from the same source could also be distinguished. CONCLUSIONS: AFLP profiling is an effective method for typing the genetically diverse organism A. butzleri. SIGNIFICANCE AND IMPACT OF THE STUDY: The study represents a comprehensive analysis of the genetic diversity of A. butzleri by use of isolates from six countries spanning three continents and also shows that several distinct A. butzleri genotypes may be found in a given environment. AFLP profiling appears to have considerable potential for molecular epidemiological studies of this ubiquitous emerging pathogen that is implicated as a causative agent of both human and animal disease. PMID- 15355538 TI - Natural community structure of cyanobacteria in the South China Sea as revealed by rpoC1 gene sequence analysis. AB - AIMS: To examine the community structure of cyanobacterial populations of the South China Sea on the surface and at depth of 80 m. METHODS AND RESULTS: Direct PCR amplification of RNA polymerase (rpoC1) genes from environmental DNAs extracted from seawater, and cloning of the fragments and sequence analysis were used. A great diversity of Prochlorococcus and Synechococcus were detected at the investigation site. Genetically related Prochlorococcus were found in both layers while Synechococcus were found only on the surface. Prochlorococcus were clustered with the known high-light adapted II genotypes, and further divided into seven groups. Synechococcus could be divided into two groups, and the second group could be further subdivided into several clades. CONCLUSION: The dominant genotype of Prochlorococcus was high-light adapted II genotype, and Synechococcus were distributed basically on the surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report about the cyanobacterial community structure in the South China Sea, and an important supplement to the current understanding of the relationship between genetic and ecological diversity and environments. PMID- 15355539 TI - Attachment and biofilm formation on stainless steel by Escherichia coli O157:H7 as affected by curli production. AB - AIMS: The aim of this study was to determine the role of curli in attachment and biofilm formation by Escherichia coli O157:H7 on stainless steel. METHODS AND RESULTS: Three curli-deficient strains (43895-, 43894- and E0018-) and three curli over-producing strains (43895+, 43894+ and E0018+) of E. coli O157:H7 were studied. Stainless steel coupons (SSC) were immersed in cell suspensions of each strain for 24 h at 4 degrees C. The number of cells attached to SSC was determined. To determine the ability of attached cells to form biofilm, SSC were immersed in 10% of tryptic soya broth up to 6 days at 22 degrees C. Curli deficient and curli-producing strains did not differ in their ability to attach to SSC, but only curli-producing strains formed biofilms. CONCLUSIONS: Curli production by E. coli O157:H7 does not affect attachment of cells on stainless steel but curli-producing strains are better able to form biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: Curli production by E. coli O157:H7 enhances its ability to form biofilm on stainless steel, thereby potentially resulting in increased difficulty in removing or killing cells by routine cleaning and sanitizing procedures used in food-processing plants. PMID- 15355540 TI - Response surface methodology for optimizing the fermentation medium of Clostridium butyricum. AB - AIMS: Strains of Clostridium butyricum have been increasingly used as probiotics for both animals and humans. The aim of this study was to develop a growth medium for cultivating C. butyricum ZJUCB using a statistical methodology. METHODS AND RESULTS: Response surface methodology (RSM) was used to evaluate the effects of variables, namely the concentrations of the glucose, pectin, soyabean cake extract, casein, corn steep flour, ammonium sulphate, sodium bicarbonate and the medium initial pH. A fractional factorial design was applied to study the main factors that affected the growth of a probiotic strain of C. butyricum currently preserved in our lab and the central composite experimental design was adopted to derive a statistical model for optimizing the composition of the fermentation medium. The experimental results showed that the optimum fermentation medium for the growth of C. butyricum was composed of 2% glucose (w/v), 0.5% pectin (w/v), 0.2% casein (w/v), 3.98% soyabean cake extract, 0.1% (NH4)2SO4 (w/v), 0.124% NaHCO3 (w/v), 0.37% corn steep flour (w/v), 0.02% MnSO4 H2O (w/v), 0.02% MgSO4 7H2O (w/v) and 0.002% CaCl2 (w/v) at pH 7.5. CONCLUSIONS: After incubating 24 h in the optimum fermentation medium, the populations of the viable organisms were estimated to be 10(9) CFU ml(-1). In the present study, we report the optimization of a growth medium that produced increased yields using statistical approach. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of bacteria as a probiotic is showing increasing potential. The development of a growth medium that has a high yield is an obvious need, and the approach to optimizing a growth medium is innovative. PMID- 15355541 TI - Influence of cultivating conditions on the alpha-galactosidase biosynthesis from a novel strain of Penicillium sp. in solid-state fermentation. AB - AIMS: The work is intended to achieve optimum culture conditions of alpha galactosidase production by a mutant strain Penicillium sp. in solid-state fermentation (SSF). METHODS AND RESULTS: Certain fermentation parameters involving incubation temperature, moisture content, initial pH value, inoculum and load size of medium, and incubation time were investigated separately. The optimal temperature and moisture level for alpha-galactosidase biosynthesis was found to be 30 degrees C and 50%, respectively. The range of pH 5.5-6.5 was favourable. About 40-50 g of medium in 250-ml flask and inoculum over 1.0 x 10(6) spores were suitable for enzyme production. Seventy-five hours of incubation was enough for maximum alpha-galactosidase production. Substrate as wheat bran supplemented with soyabean meal and beet pulp markedly improved the enzyme yield in trays. CONCLUSIONS: Under optimum culture conditions, the alpha-galactosidase activity from Penicillium sp. MAFIC-6 indicated 185.2 U g(-1) in tray of SSF. SIGNIFICANT AND IMPACT OF THE STUDY: The process on alpha-galactosidase production in laboratory scale may have a potentiality of scaling-up. PMID- 15355542 TI - A real-time polymerase chain reaction-based method for rapid and specific detection of spoilage Alicyclobacillus spp. in apple juice. AB - AIMS: To develop a real-time PCR-based rapid detection method for spoilage Alicyclobacillus spp. in juice products. METHODS AND RESULTS: The squalene-hopene cyclase-encoding gene was targeted for primer-and-probe development. Gene fragments from representative strains were cloned, and PCR primers and probe were designed by DNA sequence comparison. Selected bacteria were examined for cross reactivity by the new method. Cells were serially diluted in apple juice and saline, and examined by the new method to establish detection sensitivity. Using the newly developed Taqman real-time PCR-based method, strains of Alicyclobacillus acidocaldarius and A. acidoterrestris were detected without cross reactivity with other common food-borne micro-organisms. Detection of <10 cells per PCR reaction from juice samples was accomplished within 3-5 h. CONCLUSION: This is the first reported real-time PCR-based detection method for Alicyclobacillus spp. and its application in juice products is demonstrated. SIGNIFICANCE AND IMPACT OF THE STUDY: As a favourable alternative for the laborious and time-consuming culture- or biochemical characterization-based techniques, the system has great potential for industrial applications from raw material screening to final product quality control. PMID- 15355543 TI - Evaluation of the effectiveness of a commercially available defined substrate medium and enumeration system for measuring Escherichia coli numbers in faeces and soil samples. AB - AIMS: To determine if a commercially available defined substrate medium and enumeration system could be utilized as an effective and accurate means of enumerating Escherichia coli in environmental samples containing faeces and soil. METHODS AND RESULTS: The samples tested were either inoculated with laboratory grown E. coli or natural E. coli populations in cow faeces. The number of E. coli recovered from faeces and soil samples using the defined substrate medium and enumeration system and a miniaturized MPN method (using traditional media) was compared by analysing the difference between the two methods in relation to the mean. For four of five groups of samples analysed there was no significant difference in the number of E. coli recovered by the two methods (P > 0.05). In one batch the difference was 0.30 log, which while being statistically significant (P < 0.01) was not considered to be biologically significant. CONCLUSION: The commercially available enumeration system was significantly more precise than the miniaturized MPN method (P < 0.001). SIGNIFICANCE AND IMPACT OF THE STUDY: We conclude that the commercially available defined substrate medium and enumeration system is a suitable method for the measurement of E. coli numbers in faeces and soil samples and should provide advantages of increased precision and a reduction in laboratory analysis time. PMID- 15355544 TI - Setting priorities in health care organizations: criteria, processes, and parameters of success. AB - BACKGROUND: Hospitals and regional health authorities must set priorities in the face of resource constraints. Decision-makers seek practical ways to set priorities fairly in strategic planning, but find limited guidance from the literature. Very little has been reported from the perspective of Board members and senior managers about what criteria, processes and parameters of success they would use to set priorities fairly. DISCUSSION: We facilitated workshops for board members and senior leadership at three health care organizations to assist them in developing a strategy for fair priority setting. Workshop participants identified 8 priority setting criteria, 10 key priority setting process elements, and 6 parameters of success that they would use to set priorities in their organizations. Decision-makers in other organizations can draw lessons from these findings to enhance the fairness of their priority setting decision-making. SUMMARY: Lessons learned in three workshops fill an important gap in the literature about what criteria, processes, and parameters of success Board members and senior managers would use to set priorities fairly. PMID- 15355545 TI - Non-invasive monitoring of diaphragmatic timing by means of surface contact sensors: an experimental study in dogs. AB - BACKGROUND: Non-invasive monitoring of respiratory muscle function is an area of increasing research interest, resulting in the appearance of new monitoring devices, one of these being piezoelectric contact sensors. The present study was designed to test whether the use of piezoelectric contact (non-invasive) sensors could be useful in respiratory monitoring, in particular in measuring the timing of diaphragmatic contraction. METHODS: Experiments were performed in an animal model: three pentobarbital anesthetized mongrel dogs. The motion of the thoracic cage was acquired by means of a piezoelectric contact sensor placed on the costal wall. This signal is compared with direct measurements of the diaphragmatic muscle length, made by sonomicrometry. Furthermore, to assess the diaphragmatic function other respiratory signals were acquired: respiratory airflow and transdiaphragmatic pressure. Diaphragm contraction time was estimated with these four signals. Using diaphragm length signal as reference, contraction times estimated with the other three signals were compared with the contraction time estimated with diaphragm length signal. RESULTS: The contraction time estimated with the TM signal tends to give a reading 0.06 seconds lower than the measure made with the DL signal (-0.21 and 0.00 for FL and DP signals, respectively), with a standard deviation of 0.05 seconds (0.08 and 0.06 for FL and DP signals, respectively). Correlation coefficients indicated a close link between time contraction estimated with TM signal and contraction time estimated with DL signal (a Pearson correlation coefficient of 0.98, a reliability coefficient of 0.95, a slope of 1.01 and a Spearman's rank-order coefficient of 0.98). In general, correlation coefficients and mean and standard deviation of the difference were better in the inspiratory load respiratory test than in spontaneous ventilation tests. CONCLUSION: The technique presented in this work provides a non-invasive method to assess the timing of diaphragmatic contraction in canines, using a piezoelectric contact sensor placed on the costal wall. PMID- 15355546 TI - The efficacy of duloxetine: a comprehensive summary of results from MMRM and LOCF_ANCOVA in eight clinical trials. AB - BACKGROUND: A mixed-effects model repeated measures approach (MMRM) was specified as the primary analysis in the Phase III clinical trials of duloxetine for the treatment of major depressive disorder (MDD). Analysis of covariance using the last observation carried forward approach to impute missing values (LOCF_ANCOVA) was specified as a secondary analysis. Previous research has shown that MMRM and LOCF_ANCOVA yield identical endpoint results when no data are missing, while MMRM is more robust to biases from missing data and thereby provides superior control of Type I and Type II error compared with LOCF_ANCOVA. We compared results from MMRM and LOCF_ANCOVA analyses across eight clinical trials of duloxetine in order to investigate how the choice of primary analysis may influence interpretations of efficacy. METHODS: Results were obtained from the eight acute-phase clinical trials that formed the basis of duloxetine's New Drug Application for the treatment of MDD. All 202 mean change analyses from the 20 rating scale total scores and subscales specified a priori in the various protocols were included in the comparisons. RESULTS: In 166/202 comparisons (82.2%), MMRM and LOCF_ANCOVA agreed with regard to the statistical significance of the differences between duloxetine and placebo. In 25/202 cases (12.4%), MMRM yielded a significant difference when LOCF_ANCOVA did not, while in 11/202 cases (5.4%), LOCF_ANCOVA produced a significant difference when MMRM did not. In 110/202 comparisons (54.4%) the p-value from MMRM was lower than that from LOCF_ANCOVA, while in 69/202 comparisons (34.2%), the p-value from LOCF_ANCOVA was lower than that from MMRM. In the remaining 23 comparisons (11.4%), the p-values from LOCF_ANCOVA and MMRM were equal when rounded to the 3rd decimal place (usually as a result of both p-values being <.001). For the HAMD17 total score, the primary outcome in all studies, MMRM yielded 9/12 (75%) significant contrasts, compared with 6/12 (50%) for LOCF_ANCOVA. The expected success rate was 80%. CONCLUSIONS: Important differences exist between MMRM and LOCF_ANCOVA. Empirical research has clearly demonstrated the theoretical advantages of MMRM over LOCF_ANCOVA. However, interpretations regarding the efficacy of duloxetine in MDD were unaffected by the choice of analytical technique. PMID- 15355547 TI - Comparison of glucose tolerance in renal transplant recipients and hemodialysis patients. AB - BACKGROUND: Impaired glucose tolerance is a risk factor for atherosclerosis in hemodialysis patients and renal transplant recipients. METHODS: To check the relationship of impaired glucose tolerance with the other atherosclerotic risk factors, fasting blood sugar and the standard two hour glucose tolerance test, serum tryglyceride, serum cholesterol, cyclosporine through level (in renal transplant recipients) and hemoglobin A1C were measured in 55 stable renal transplant recipients, 55 hemodialysis patients and 55 healthy controls with similar demographic characteristics. Patients with diabetes mellitus and propranolol consumers were excluded. The mean age and female to male ratio were 39 +/- 7 years and 23/22, respectively. RESULTS: Four of the renal transplant recipients and twelve of the hemodialysis patients had impaired glucose tolerance. Significant linear correlation was observed with body mass index and IGT only in hemodialysis patients (r = 0.4, p = 0.05). Glucose tolerance also had a significant correlation with triglyceride levels (217.2 +/- 55 mg/dl in hemodialysis patients vs. 214.3 +/- 13 mg/dl in renal transplant recipients and 100.2 +/- 18 mg/dl in control groups, p = 0.001). The glucose tolerance had significant relationship with higher serum cholesterol levels only in the renal transplant recipients (269.7 +/- 54 in renal transplant recipients vs. 199.2 +/- 36.6 mg/dl in hemodialysis and 190.5 +/- 34 mg/dl in control groups, p = 0.0001). In the renal transplant recipients, a linear correlation was observed with glucose tolerance and both the serum cyclosporine level (r = 0.9, p = 0.001) and the hemoglobin A1C concentration (6.2 +/- 0.9 g/dl). The later correlation was also observed in the hemodialysis patients (6.4 +/- 0.7 g/dl; r = 67, p = 0.001). CONCLUSIONS: We conclude that although fasting blood sugar is normal in non diabetic renal transplant and hemodialysis patients, impaired glucose tolerance could be associated with the other atherosclerotic risk factors. PMID- 15355548 TI - A new method for determination of varicella-zoster virus immunoglobulin G avidity in serum and cerebrospinal fluid. AB - BACKGROUND: Avidity determination of antigen-specific immunoglobulin G (IgG) antibodies is an established serological method to differentiate acute from past infections. In order to compare the avidity of varicella-zoster virus (VZV) IgG in pairs of serum and cerebrospinal fluid (CSF) samples, we developed a new technique of avidity testing, the results of which are not influenced by the concentration of specific IgG. METHODS: The modifications introduced for the new VZV IgG avidity method included the use of urea hydrogen peroxide as denaturing reagent, the adaptation of the assay parameters in order to increase the sensitivity for the detection of low-level VZV IgG in CSF, and the use of a new calculation method for avidity results. The calculation method is based on the observation that the relationship between the absorbance values of the enzyme immunoassays with and without denaturing washing step is linear. From this relationship, a virtual absorbance ratio can be calculated. To evaluate the new method, a panel of serum samples from patients with acute and past VZV infection was tested as well as pairs of serum and CSF. RESULTS: For the serum panel, avidity determination with the modified assay gave results comparable to standard avidity methods. Based on the coefficient of variation, the new calculation method was superior to established methods of avidity calculation. CONCLUSIONS: The new avidity method permits a meaningful comparison of VZV IgG avidity in serum and CSF and should be of general applicability for easy determination of avidity results, which are not affected by the concentration of specific IgG. PMID- 15355549 TI - Integrated analysis of metabolic phenotypes in Saccharomyces cerevisiae. AB - BACKGROUND: The yeast Saccharomyces cerevisiae is an important microorganism for both industrial processes and scientific research. Consequently, there have been extensive efforts to characterize its cellular processes. In order to fully understand the relationship between yeast's genome and its physiology, the stockpiles of diverse biological data sets that describe its cellular components and phenotypic behavior must be integrated at the genome-scale. Genome-scale metabolic networks have been reconstructed for several microorganisms, including S. cerevisiae, and the properties of these networks have been successfully analyzed using a variety of constraint-based methods. Phenotypic phase plane analysis is a constraint-based method which provides a global view of how optimal growth rates are affected by changes in two environmental variables such as a carbon and an oxygen uptake rate. Some applications of phenotypic phase plane analysis include the study of optimal growth rates and of network capacity and function. RESULTS: In this study, the Saccharomyces cerevisiae genome-scale metabolic network was used to formulate a phenotypic phase plane that displays the maximum allowable growth rate and distinct patterns of metabolic pathway utilization for all combinations of glucose and oxygen uptake rates. In silico predictions of growth rate and secretion rates and in vivo data for three separate growth conditions (aerobic glucose-limited, oxidative-fermentative, and microaerobic) were concordant. CONCLUSIONS: Taken together, this study examines the function and capacity of yeast's metabolic machinery and shows that the phenotypic phase plane can be used to accurately predict metabolic phenotypes and to interpret experimental data in the context of a genome-scale model. PMID- 15355550 TI - Evidence for positive selection on Mycobacterium tuberculosis within patients. AB - BACKGROUND: While the pathogenesis and epidemiology of tuberculosis are well studied, relatively little is known about the evolution of the infectious agent Mycobacterium tuberculosis, especially at the within-host level. The insertion sequence IS6110 is a genetic marker that is widely used to track the transmission of tuberculosis between individuals. This and other markers may also facilitate our understanding of the disease within patients. RESULTS: This article presents three lines of evidence supporting the action of positive selection on M. tuberculosis within patients. The arguments are based on a comparison between empirical findings from molecular epidemiology, and population genetic models of evolution. Under the hypothesis of neutrality of genotypes, 1) the mutation rate of the marker IS6110 is unusually high, 2) the time it takes for substitutions to occur within patients is too short, and 3) the amount of polymorphism within patients is too low. CONCLUSIONS: Empirical observations are explained by the action of positive selection during infection, or alternatively by very low effective population sizes. I discuss the possible roles of antibiotic treatment, the host immune system and extrapulmonary dissemination in creating opportunities for positive selection. PMID- 15355552 TI - Iterative class discovery and feature selection using Minimal Spanning Trees. AB - BACKGROUND: Clustering is one of the most commonly used methods for discovering hidden structure in microarray gene expression data. Most current methods for clustering samples are based on distance metrics utilizing all genes. This has the effect of obscuring clustering in samples that may be evident only when looking at a subset of genes, because noise from irrelevant genes dominates the signal from the relevant genes in the distance calculation. RESULTS: We describe an algorithm for automatically detecting clusters of samples that are discernable only in a subset of genes. We use iteration between Minimal Spanning Tree based clustering and feature selection to remove noise genes in a step-wise manner while simultaneously sharpening the clustering. Evaluation of this algorithm on synthetic data shows that it resolves planted clusters with high accuracy in spite of noise and the presence of other clusters. It also shows a low probability of detecting spurious clusters. Testing the algorithm on some well known micro-array data-sets reveals known biological classes as well as novel clusters. CONCLUSIONS: The iterative clustering method offers considerable improvement over clustering in all genes. This method can be used to discover partitions and their biological significance can be determined by comparing with clinical correlates and gene annotations. The MATLAB programs for the iterative clustering algorithm are available from http://linus.nci.nih.gov/supplement.html PMID- 15355551 TI - Experimental evaluation of the relationship between lethal or non-lethal virulence and transmission success in malaria parasite infections. AB - BACKGROUND: Evolutionary theory suggests that the selection pressure on parasites to maximize their transmission determines their optimal host exploitation strategies and thus their virulence. Establishing the adaptive basis to parasite life history traits has important consequences for predicting parasite responses to public health interventions. In this study we examine the extent to which malaria parasites conform to the predicted adaptive trade-off between transmission and virulence, as defined by mortality. The majority of natural infections, however, result in sub-lethal virulent effects (e.g. anaemia) and are often composed of many strains. Both sub-lethal effects and pathogen population structure have been theoretically shown to have important consequences for virulence evolution. Thus, we additionally examine the relationship between anaemia and transmission in single and mixed clone infections. RESULTS: Whereas there was a trade-off between transmission success and virulence as defined by host mortality, contradictory clone-specific patterns occurred when defining virulence by anaemia. A negative relationship between anaemia and transmission success was found for one of the parasite clones, whereas there was no relationship for the other. Notably the two parasite clones also differed in a transmission phenotype (gametocyte sex ratio) that has previously been shown to respond adaptively to a changing blood environment. In addition, as predicted by evolutionary theory, mixed infections resulted in increased anaemia. The increased anaemia was, however, not correlated with any discernable parasite trait (e.g. parasite density) or with increased transmission. CONCLUSIONS: We found some evidence supporting the hypothesis that there is an adaptive basis correlating virulence (as defined by host mortality) and transmission success in malaria parasites. This confirms the validity of applying evolutionary virulence theory to biomedical research and adds support to the prediction that partially effective vaccines may select for increasingly virulent malaria parasite strains. By contrast, there was no consistent correlation between transmission and sub lethal anaemia, a more common outcome of malaria infection. However, overall, the data are not inconsistent with the recent proposal that sub-lethal effects may impose an upper limit on virulence. Moreover, clone specific differences in transmission phenotypes linked to anaemia do suggest that there is considerable adaptive potential relating anaemia and transmission that may lead to uncertain consequences following intervention strategies. PMID- 15355553 TI - Boning up for health: the national bone and joint decade. PMID- 15355554 TI - Leptin, bone mass, and the thrifty phenotype. PMID- 15355555 TI - Association of the G-174C variant in the interleukin-6 promoter region with bone loss and fracture risk in older women. AB - We analyzed the association between the IL-6 G-174C polymorphism and osteoporosis phenotypes in 3376 older women. Women with the C/C genotype had a significantly slower rate of decline in hip BMD and a 33% lower risk of wrist fracture than women with the G/G genotype. Variation at the IL-6 locus may contribute to the genetic susceptibility to bone fragility. INTRODUCTION: Interleukin 6 (IL-6) promotes osteoclast formation and bone resorption. The C allele of the G-174C polymorphism in the IL-6 promoter region has been related to lower gene transcription and plasma IL-6 levels. MATERIALS AND METHODS: In this study, we evaluated the relationship between the IL-6 G-174C polymorphism and BMD, the rate of decline in BMD, and the risk of fracture in 3376 women 65 years of age and older participating in the Study of Osteoporotic Fractures. BMD was measured at the distal and proximal radius using single photon absorptiometry and at the hip using DXA. Hip BMD was measured again an average of 3.5 years later. Incident fractures over an average of 10.8 years of follow-up were confirmed by physician adjudication of radiology reports. RESULTS: Distal and proximal radius BMD was lowest among women with the G/G genotype, intermediate in the heterozygotes, and highest in women with the C/C genotype (p = 0.016 and p = 0.049, respectively), although the differences between the genotypes were small. While there were no differences by genotype with initial hip BMD, women with the C/C genotype experienced a slower rate of decline in total hip and femoral neck BMD compared with women with the G/G genotype (p = 0.004 and p = 0.029, respectively). Women with the C/C genotype also had 33% lower risk of wrist fracture compared with women with the G/G genotype, independent of age, body mass index, estrogen use, and study center (RR, 0.67; 95% CI, 0.45, 1.00; p = 0.048), whereas heterozygous women had a more intermediate risk (RR, 0.85; 95% CI, 0.65, 1.12; p = 0.256). No association was found between genotype and risk of hip or all non-spine fractures. CONCLUSIONS: These results suggest that the IL-6 G-174C promoter polymorphism may be a genetic marker for bone loss and wrist fracture among older women. PMID- 15355556 TI - Influence of LRP5 polymorphisms on normal variation in BMD. AB - Genetic studies based on cohorts with rare and extreme bone phenotypes have shown that the LRP5 gene is an important genetic modulator of BMD. Using family-based and case-control approaches, this study examines the role of the LRP5 gene in determining normal population variation of BMD and describes significant association and suggestive linkage between LRP5 gene polymorphisms and BMD in >900 individuals with a broad range of BMD. INTRODUCTION: Osteoporosis is a common, highly heritable condition determined by complex interactions of genetic and environmental etiologies. Genetic factors alone can account for 50-80% of the interindividual variation in BMD. Mutations in the LRP5 gene on chromosome 11q12 13 have been associated with rare syndromes characterized by extremely low or high BMD, but little is known about the contribution of this gene to the development of osteoporosis and determination of BMD in a normal population. MATERIALS AND METHODS: To examine the entire spectrum of low to high BMD, 152 osteoporotic probands, their families (597 individuals), and 160 women with elevated BMD (T score > 2.5) were recruited. BMD at the lumbar spine, femoral neck, and hip were measured in each subject using DXA. RESULTS: PAGE sequencing of the LRP5 gene revealed 10 single nucleotide polymorphisms (SNPs), 8 of which had allele frequencies of >5%, in exons 8, 9, 10, 15, and 18 and in introns 6, 7, and 21. Within families, a strong association was observed between an SNP at nucleotide C171346A in intron 21 and total hip BMD (p < 1 x 10(-5) in men only, p = 0.0019 in both men and women). This association was also observed in comparisons of osteoporotic probands and unrelated elevated BMD in women (p = 0.03), along with associations with markers in exons 8 (C135242T, p = 0.007) and 9 (C141759T, p = 0.02). Haplotypes composed of two to three of the SNPs G121513A, C135242T, G138351A, and C141759T were strongly associated with BMD when comparing osteoporotic probands and high BMD cases (p < 0.003). An SNP at nucleotide C165215T in exon 18 was linked to BMD at the lumbar spine, femoral neck, and total hip (parametric LOD scores = 2.8, 2.5, and 2.2 and nonparametric LOD scores = 0.3, 1.1, and 2.2, respectively) but was not genetically associated with BMD variation. CONCLUSION: These results show that common LRP5 polymorphisms contribute to the determination of BMD in the general population. PMID- 15355557 TI - Bone remodeling increases substantially in the years after menopause and remains increased in older osteoporosis patients. AB - Bone remodeling rates (Ac.f) were measured in transilial biopsy specimens from 50 healthy premenopausal women before and 1 year after menopause, in 34 healthy women 13 years past menopause, and in 89 women with untreated osteoporosis. Ac.f nearly doubled 1 year after menopause, tripled 13 years after menopause, and remained elevated in women with osteoporosis. INTRODUCTION: Increased bone remodeling rates are associated with increased skeletal fragility independent of bone mass, partially accounting for the age-related increase in fracture risk in women that is independent of bone loss. We examined bone remodeling rates before and after menopause and in women with osteoporosis by measurements of activation frequency (Ac.f, #/year) in transilial bone biopsy specimens. MATERIALS AND METHODS: We recruited 75 women, > 46 years old, who had premenopausal estradiol and gonadotropin levels and regular menses. During 9.5 years of observation, 50 women experienced normal menopause and had 2 transilial bone biopsy specimens after tetracycline labeling, one at the beginning of observation and the second 12 months after the last menses, when serum follicle-stimulating hormone (FSH) was > 75 mIU/ml and serum estradiol was < 20 pg/ml. Ac.f was also computed for a group of older healthy postmenopausal women and a group of women with untreated osteoporosis studied earlier by the same biopsy (Bx) and labeling protocol. RESULTS: Median Ac.f rose from 0.13/year to 0.24/year (p < 0.001) across menopause and was greater still in the older normals (p < 0.008) than in the second Bx. Ac.f was not significantly greater in the osteoporosis patients than in the older postmenopausal normals. CONCLUSION: Bone remodeling rates double at menopause, triple 13 years later, and remain elevated in osteoporosis. This change contributes to increases in age-related skeletal fragility in women. PMID- 15355558 TI - Physical activity and calcium consumption are important determinants of lower limb bone mass in older women. AB - A population-based study of 1363 older women showed that the 24% who achieved high physical activity and dietary calcium intakes had a 5.1% higher hip BMD than those who did not, supporting the concept that lifestyle factors play an important role in the maintenance of lower extremity bone mass in older women. INTRODUCTION: Although there is general agreement that increased dietary calcium consumption and exercise can slow bone loss in older women, the amount required to have this effect in an older population remains uncertain. This study was devised to examine the effects of calcium consumption (CC) and physical activity (PA) (lifestyle management) on bone mass in an older female population. MATERIALS AND METHODS: Using a cross-sectional study design, a population-based sample of older women (mean age, 75 +/- 3 years) had hip and heel bone mass measured using DXA (Hologic 4500A; n = 1076) and quantitative ultrasound (QUS, Lunar Achilles; n = 1363), respectively. CC and PA were measured by a validated habitual food frequency and activity questionnaire, respectively. Dose-response effects of PA and CC on bone mass were examined using ANOVA. RESULTS AND CONCLUSIONS: Division of the PA and CC into tertiles best described the dose-response effects. After adjustment for CC, age, weight, alcohol consumption, and cigarette smoking, high PA compared with medium or low PA was associated with higher hip BMD and heel QUS (total hip BMD, 3.1%; p < 0.001; QUS stiffness, 2.7%; p = 0.002). After adjustment for PA and covariates, high or medium CC compared with low CC was associated with higher total hip BMD (1.8%; p = 0.027), with no effect at the QUS heel site. PA and CC were dichotomized at the cut-points for effects on BMD. The combination of high PA and CC, achieved by 24% of the population, was associated with a total hip BMD that was 5.1% higher (34% of SD) than those individuals in the low PA and CC group. Stiffness was 3.6% (23% of SD) higher in the high PA and CC group than in the low PA and CC group. If the whole population undertook and achieved a high PA and high CC lifestyle, the population risk of hip fractures may be expected to be reduced by about 17% in this age group as a result of beneficial effects on the musculoskeletal system. PMID- 15355559 TI - An improved segmentation method for in vivo microCT imaging. AB - Image segmentation methods for microCT can influence the accuracy of bone morphometry calculations. A new automated segmentation method is introduced, and its performance is compared with standard segmentation methods. The new method can improve the results of in vivo microCT, where the need to keep radiation dose low limits scan quality. INTRODUCTION: An important topic for microCT analysis of bone samples is the segmentation of the original reconstructed grayscale data sets to separate bone from non-bone. Problems like noise, resolution limitations, and beam-hardening make this a nontrivial issue. Inappropriate segmentation methods will reduce the potential power of microCT and may introduce bias in the architectural measurements, in particular, when new in vivo microCT with its inherent limitations in scan quality is used. Here we introduce a new segmentation method using local thresholds and compare its performance to standard global segmentation methods. MATERIAL AND METHODS: The local threshold method was validated by comparing the result of the segmentation with histology. Furthermore, the effect of choosing this new method versus standard segmentation methods using global threshold values was investigated by studying the sensitivity of these methods to signal to noise ratio and resolution. RESULTS: Using the new method on high-quality scans yielded accurate results and virtually no differences between histology and the segmented data sets could be observed. When prior knowledge about the volume fraction of the bone was available the global threshold also resulted in appropriate results. Degrading the scan quality had only minor effects on the performance of the new segmentation method. Although global segmentation methods were not sensitive to noise, it was not possible to segment both lower mineralized thin trabeculae and the higher mineralized cortex correctly with the same threshold value. CONCLUSION: At high resolutions, both the new local and conventional global segmentation methods gave near exact representations of the bone structure. When scanned samples are not homogenous (e.g., thick cortices and thin trabeculae) and when resolution is relatively low, the local segmentation method outperforms global methods. It maximizes the potential of in vivo microCT by giving good structural representation without the need to use longer scanning times that would increase absorption of harmful X-ray radiation by the living tissue. PMID- 15355560 TI - Identification of biphenylcarboxylic acid derivatives as a novel class of bone resorption inhibitors. AB - A novel class of biphenylcarboxylic acid derivatives are described that inhibit osteoclastic bone resorption in vitro by promoting osteoclast apoptosis and that prevent ovariectomy-induced bone loss in vivo. The compounds act by a novel mechanism that seems to be distinct from existing antiresorptive drugs. INTRODUCTION: Many common bone diseases such as osteoporosis, Paget's disease, and cancer-associated bone disease are characterized by excessive bone loss caused by increased osteoclastic activity. Successful treatment of these diseases is based on osteoclast inhibition. The osteoclast inhibitory drugs that are currently available fall into relatively few mechanistic classes, indicating the need to identify novel antiresorptives. Here we describe a series of biphenylcarboxylic acid derivatives that have potent inhibitory effects on osteoclastic bone resorption in vitro and on ovariectomy-induced bone loss in vivo. MATERIALS AND METHODS: Compounds were tested for inhibitory effects on bone resorption in vitro using mouse osteoblast-bone marrow co-cultures, isolated rabbit osteoclasts, and mouse osteoclasts generated from bone marrow. Some experiments were also performed on human osteoclasts generated from peripheral blood mononuclear cells. We also investigated the effects of specific compounds on ovariectomy-induced bone loss in vivo in mice. RESULTS: One of the most potent compounds identified was the butanediol ester of biphenyl carboxylic acid (ABD056), which inhibited osteoclast formation in mouse osteoblast-bone marrow co cultures by 50% (IC50) at a concentration of 26 microM and in macrophage-colony stimulating factor (M-CSF)- and RANKL-stimulated mouse bone marrow cultures with an IC50 of 8 microM. Mechanistic studies showed that ABD056 caused osteoclast apoptosis and inhibited TNFalpha-induced NF-kappaB activation. No inhibitory effects on osteoblast growth or differentiation were observed at concentrations of up to 100 microM. When administered to mice at doses of 5 and 10 mg/kg/day, ABD056 prevented ovariectomy-induced bone loss. CONCLUSIONS: Butanediol biphenylcarboxylic acid derivatives represent a new class of antiresorptive drug that might be of therapeutic value in the prevention and treatment of diseases characterized by osteoclast activation such as osteoporosis, cancer-associated bone disease, and Paget's disease of bone. PMID- 15355561 TI - Unmasking the osteoinductive effects of a G-protein-coupled receptor (GPCR) kinase (GRK) inhibitor by treatment with PTH(1-34). AB - The effects of GPCR systems in bone are regulated by a family of enzymes termed GRKs. We found that (1) GRK inhibition in osteoblasts has age-dependent effects on bone mass, and (2) the anabolic actions of GRK inhibition are revealed by treatment with PTH(1-34). INTRODUCTION: The effects of G-protein-coupled receptor (GPCR) systems in bone are modulated by a family of enzymes termed GPCR kinases (GRKs). These enzymes directly phosphorylate GPCR substrate and desensitize receptor signaling. We previously found that expression of a GRK inhibitor in osteoblasts using transgenic (TG) technologies enhanced bone remodeling, and in turn, increased BMD in 6-week-old TG mice compared with non-TG littermate controls, presumably because of enhanced GPCR function. The aim of this study was to determine the age-dependent effects of the transgene. MATERIALS AND METHODS: BMD was monitored in TG mice and in controls at 6-week, 3-month, and 6-month time points. To determine if the transgene enhanced responsiveness of bone to parathyroid hormone (PTH), we measured cyclic adenosine monophosphate (cAMP) generation by mouse calvaria ex vivo as well as the effects of treatment with PTH(1-34) on BMD, bone histomorphometry, and expression of the PTH-responsive gene RANKL in both TG mice and non-TG controls. RESULTS: Consistent with our previous findings, we found that BMD was increased in TG mice compared with controls at 6 weeks of age. The increase in BMD was most prominent in trabecular rich lumbar spine and was not observed in cortical bone of the femoral shaft. In contrast to younger animals, however, BMD in older TG mice was not statistically different compared with non-TG mice at 3 months of age and was similar to non-TG animals at 6 months of age. The GRK inhibitor seemed to promote GPCR activation in older mice, however, because (1) PTH-induced cAMP generation by mouse calvaria ex vivo was enhanced in TG mice compared with controls, (2) GRK inhibition increased responsiveness of lumbar spine to the osteoinductive actions of PTH(1 34), and (3) the enhanced anabolic effect of PTH(1-34) was associated with increased expression of the PTH-responsive gene RANKL in calvaria of the TG animals. Bone histomorphometry confirmed that PTH(1-34) increased trabecular bone volume in TG mice and found that this increase in bone mass was caused by enhanced bone formation, predominantly as a result of an increase in the mineral apposition rate (MAR). CONCLUSIONS: These data suggest that the anabolic effects of GRK inhibition are age dependent. The osteoinductive actions of the GRK inhibitor are, however, unmasked by treatment with PTH(1-34). PMID- 15355562 TI - Possible roles of Runx1 and Sox9 in incipient intramembranous ossification. AB - We evaluated the detailed expression patterns of Runx1 and Sox9 in various types of bone formation, and determined whether Runx1 expression was affected by Runx2 deficiency and Runx2 expression by Runx1 deficiency. Our results indicate that both Runx1 and Sox9 are intensely expressed in the future osteogenic cell compartment and in cartilage. The pattern of Runx1 and Sox9 expression suggests that both genes could potentially be involved in incipient intramembranous bone formation during craniofacial development. INTRODUCTION: Runx1, a gene essential for hematopoiesis, contains RUNX binding sites in its promoter region, suggesting possible cross-regulation with Runx2 and potential regulatory roles in bone development. On the other hand, Sox9 is essential for chondrogenesis, and haploinsufficiency of Sox9 leads to premature ossification of the skeletal system. In this study, we studied the possible roles of Runx1 and Sox9 in bone development. MATERIALS AND METHODS: Runx1, Runx2/Osf2, and Sox9 expression was evaluated by in situ hybridization in the growing craniofacial bones of embryonic day (E)12-16 mice and in the endochondral bone-forming regions of embryonic and postnatal long bones. In addition, we evaluated Runx2/Osf2 expression in the growing face of Runx1 knockout mice at E12.5 and Runx1 expression in Runx2 knockout mice at E14.5. RESULTS: Runx1 and Sox9 were expressed in cartilage, and the regions of expression expanded to the neighboring Runx2-expressing osteogenic regions. Expression of both Runx1 and Sox9 was markedly downregulated on ossification. Runx1 and Sox9 expression was absent in the regions of endochondral bone formation and in actively modeling or remodeling bone tissues in the long bones as well as in ossified craniofacial bones. Runx2 expression was not affected by gene disruption of Runx1, whereas the expression domains of Runx1 were extended in Runx2(-/-) mice compared with wildtype mice. CONCLUSIONS: Runx1 and Sox9 are specifically expressed in the osteogenic cell compartments in the craniofacial bones and the bone collar of long bones, and this expression is downregulated on terminal differentiation of osteoblasts. Our results suggest that Runx1 may play a role in incipient intramembranous bone formation. PMID- 15355563 TI - Targeted expression of SHH affects chondrocyte differentiation, growth plate organization, and Sox9 expression. AB - The role of Hedgehogs (Hh) in murine skeletal development was studied by overexpressing human Sonic Hedgehog (SHH) in chondrocytes of transgenic mice using the collagen II promoter/enhancer. Overexpression caused a lethal craniorachischisis with major alterations in long bones because of defects in chondrocyte differentiation. INTRODUCTION: Hedgehogs (Hhs) are a family of secreted polypeptides that play important roles in vertebrate development, controlling many critical steps of cell differentiation and patterning. Skeletal development is affected in many different ways by Hhs. Genetic defects and anomalies of Hhs signaling pathways cause severe abnormalities in the appendicular, axial, and cranial skeleton in man and other vertebrates. MATERIALS AND METHODS: Genetic manipulation of mouse embryos was used to study in vivo the function of SHH in skeletal development. By DNA microinjection into pronuclei of fertilized oocytes, we have generated transgenic mice that express SHH specifically in chondrocytes using the cartilage-specific collagen II promoter/enhancer. Transgenic skeletal development was studied at different embryonic stages by histology. The expression pattern of specific chondrocyte molecules was studied by immunohistochemistry and in situ hybridization. RESULTS: Transgenic mice died at birth with severe craniorachischisis and other skeletal defects in ribs, sternum, and long bones. Detailed analysis of long bones showed that chondrocyte differentiation was blocked at prehypertrophic stages, hindering endochondral ossification and trabecular bone formation, with specific defects in different limb segments. The growth plate was highly disorganized in the tibia and was completely absent in the femur and humerus, leading to skeletal elements entirely made of cartilage surrounded by a thin layer of bone. In this cartilage, chondrocytes maintained a columnar organization that was perpendicular to the bone longitudinal axis and directed toward its outer surface. The expression of SHH receptor, Patched-1 (Ptc1), was greatly increased in all cartilage, as well as the expression of parathyroid hormone-related protein (PTHrP) at the articular surface; while the expression of Indian Hedgehog (Ihh), another member of Hh family that controls the rate of chondrocyte maturation, was greatly reduced and restricted to the displaced chondrocyte columns. Transgenic mice also revealed the ability of SHH to upregulate the expression of Sox9, a major transcription factor implicated in chondrocyte-specific gene expression, in vivo and in vitro, acting through the proximal 6.8-kb-long Sox9 promoter. CONCLUSION: Transgenic mice show that continuous expression of SHH in chondrocytes interferes with cell differentiation and growth plate organization and induces high levels and diffuse expression of Sox9 in cartilaginous bones. PMID- 15355564 TI - Malignant autosomal recessive osteopetrosis caused by spontaneous mutation of murine Rank. AB - We report the first case of lethal autosomal recessive osteopetrosis in mice caused by a spontaneous 8-bp deletion in exon 2 of the Rank gene. The phenotype, including a block in RANKL-dependent osteoclast differentiation and lymph node agenesis, copies that of Rank(-/-) mice, which have been produced by targeted recombination. INTRODUCTION: Commitment of osteoclast progenitors to the osteoclast lineage requires RANKL/RANK-mediated intercellular signals. Gene targeted defects in this signaling pathway resulted in osteoclast deficiency and severe osteopetrosis in mice, but to date, there have been no reports of spontaneous mutations in Rankl or Rank resulting in osteopetrosis. MATERIALS AND METHODS: Mice with malignant osteopetrosis and absent lymph nodes appeared spontaneously in a highly inbred colony. Appropriate crosses were analyzed to establish the pattern of inheritance. Tissues from affected pups and littermates were evaluated grossly, histopathologically, and radiographically. Osteoclast development from splenocytes was tested in vitro under a variety of conditions, including after infection with RANK-encoding retrovirus. Rank mutational analysis was performed by direct sequencing of RT-PCR products and genomic DNA. RESULTS: The inheritance pattern was consistent with autosomal recessive inheritance, and the phenotype resembled that of either Rankl or Rank knockout mice with the exception of as yet unexplained death of most mice 2-3 weeks after weaning. Osteoclast precursors from the spleens of affected pups failed to form osteoclasts in vitro when stimulated with macrophage-colony stimulating factor (M CSF) and RANKL, unless they were forced to express wildtype Rank cDNA. Molecular genetic studies identified an 8-bp deletion in exon 2 of the Rank gene. The resulting allele, termed Rank(del8), encodes only a small portion of the RANK extracellular domain, which is probably nonfunctional. CONCLUSIONS: The phenotypic similarities between Rank(del8) and mice previously described with a combined insertion and deletion in Rank confirm the role of this receptor in osteoclastogenesis and lymph node development and suggest that some forms of malignant osteopetrosis in humans could result from a similar defect. PMID- 15355565 TI - Transient retention of endochondral cartilaginous matrix with bisphosphonate treatment in a long-term rabbit model of distraction osteogenesis. AB - Bisphosphonates induce major increases in strength of callus in distraction osteogenesis in the short term. Poor understanding of the underlying mechanism, however, raises concerns about long-term consequences. In this long-term study in 32 rabbits, zoledronic acid transiently increased trabeculae by delayed temporal progression of endochondral bone remodeling but did not prevent radiographic completion of bone repair. INTRODUCTION: We hypothesized that bisphosphonate inhibition of osteoclast-mediated resorption would retain bone during repair, producing a larger callus in the short term. However, if remodeling was not restored, completion of the bone repair process in the long term could be jeopardized. MATERIALS AND METHODS: Juvenile rabbits underwent right tibial osteotomy and 2 weeks of distraction, followed by a period of consolidation. Animals received saline (controls) or zoledronic acid (ZA; 0.1 mg/kg at surgery and again 2 weeks later), and distracted tibias were examined by radiograph, DXA, histology, and histomorphometry at 2, 4, 6, 18, and 44 weeks after surgery. RESULTS: Regenerated bone in ZA-treated animals was denser than controls on radiographs at 6 weeks and had more distinct radiodense trabeculae and retention of original cortices at 18 weeks. By 44 weeks, controls and ZA-treated animals were radiographically healed and indistinguishable. Regenerate BMD and BMC increased between 2 and 4 weeks in all animals, with a greater effect in ZA. At 6 weeks, BMD and BMC in ZA-treated animals were 1.6- and 2-fold greater, respectively, than controls (p < 0.01). From 6 to 44 weeks, the control values gradually increased and approached the ZA-treated values. Regenerate bone volume and trabecular number by histomorphometry were from 1.6- to 2-fold greater in ZA treated animals at 6 and 18 weeks (p < 0.05). Endochondral cartilaginous matrix volume was up to 2.4-fold greater in ZA-treated animals at 2 and 4 weeks (p < 0.05). TRACP+ cells in ZA-treated animals were larger with more nuclei. Mineral apposition rate and osteoblast number and surface were lower in ZA-treated animals at 6 weeks (p < 0.01) but not at later times. CONCLUSIONS: Disruption of TRACP+ cell function by ZA during bone regeneration seems to lead to an accretion of cancellous bone built on a larger endochondral cartilaginous matrix and increased bone mass, consistent with reported increases in short-term callus strength. This increase in bone mass, caused by a delay in remodeling, provided a transient advantage without preventing radiographic completion of the bone repair process in the long term. Noncontinuous treatment with nitrogen-containing bisphosphonates thus can have short-term beneficial effects without preventing long-term bone repair. PMID- 15355566 TI - Osteopontin-deficiency suppresses growth of B16 melanoma cells implanted in bone and osteoclastogenesis in co-cultures. AB - Tumor metastasis and invasion to bone is one of major medical issues in our modern societies. Osteopontin deficiency decreased tumor invasion in bone based on knockout mouse study. In bone, osteopontin is a positive factor to increase tumor invasion. INTRODUCTION: Osteopontin is an arginine-glycine-aspartate (RGD) containing protein and is recognized by integrin family members. Osteopontin promotes cell attachment to bone, where it is abundantly present. Because osteopontin levels were reported to be elevated in patients bearing highly metastatic tumors, this molecule has been implicated in the metastasis of tumors. However, the effect of osteopontin on the invasion of tumor cells in bone microenvironment has not been clear. The purpose of this paper is to elucidate the effect of host osteopontin on the behavior of tumor cells in bone. MATERIALS AND METHODS: Bone marrow ablation was conducted in the femora of mice, and B16 melanoma cells were injected directly into the ablated bone marrow space of the osteopontin-deficient and wildtype mice. RESULT: Invasion foci of B16 melanoma cells in the cortical bone was observed 7 weeks after tumor cell implantation. The number of the foci was 5-fold less in osteopontin-deficient mice compared with that in wildtype mice. In wildtype mice, trabecular bone formation was not observed in the ablated marrow space where tumor cells were injected. In contrast, significant levels of trabecular bone were observed in the marrow space of osteopontin-deficient mice even after tumor cells were injected. To examine cellular mechanisms underlying these observations, co-cultures of bone marrow cells and B16 cells were conducted. While the presence of B16 cells promoted TRACP+ cell development in wildtype bone marrow cells, such enhancement in TRACP+ cell formation by the co-cultures with B16 cells was reduced in the case of bone marrow cells from osteopontin-deficient mice. CONCLUSIONS: Osteopontin deficiency reduced the bone loss caused by tumor cell implantation into the bone marrow space. PMID- 15355567 TI - Human bone marrow stromal cells protect prostate cancer cells from TRAIL-induced apoptosis. AB - Tumor-derived OPG has recently been shown to protect prostate cancer cells from apoptosis. This study has confirmed that bone marrow stromal cell-derived OPG also suppresses cytokine-induced apoptosis in this tumor type, suggesting that it may be the presence of bone-derived OPG that is responsible for the observed preference of these cells in colonizing the skeleton. INTRODUCTION: Metastasis to the skeleton occurs in around 70% of patients with advanced prostate cancer (CaP), suggesting that the bone microenvironment may provide factors that favor the growth and survival of prostate cancer cells. Osteoprotegerin (OPG) is a molecule involved in bone remodeling, where it acts as an inhibitor of osteoclastogenesis, but it is also a decoy receptor for TRAIL/Apo 2L, a member of the TNF family of pro-apoptotic cytokines. The aim of this study was to determine whether OPG produced by human bone marrow stromal cells could protect prostate cancer cells from TRAIL-induced apoptosis. MATERIALS AND METHODS: Human bone marrow stromal cell cultures were generated from bone biopsies taken from newly diagnosed untreated CaP patients with (M1) or without (M0) bony metastasis. The stromal origin of these cells was confirmed by Western blot analysis using antibodies raised to stromal and epithelial markers. Media were conditioned over the cultures of these cells for 4 days, and levels of OPG were determined using an ELISA. The human prostate cancer cell line PC3 was challenged with TRAIL (50 ng/ml) in fresh media or in media supplemented with 50% conditioned media, and apoptosis was assessed using DAPI stain. The effects of specific removal of OPG activity by immunoprecipitation or by co-treatment of cultures with an alternative ligand for OPG (RANKL) were also tested. RESULTS AND CONCLUSIONS: The presence of stromal cell conditioned media in PC3 culture significantly reduced TRAIL-induced apoptosis. All stromal cell lines isolated were shown to express OPG and to release this protein into the conditioned media. Immunoprecipitation of OPG and co-treatment of cultures with sRANKL reversed the protective effects of the conditioned media. These data suggest that at least part of the survival advantage gained by CaP cells in colonizing bone may be caused by the production of OPG by tumor-associated stromal cells. PMID- 15355568 TI - (Pre-)osteoclasts induce retraction of osteoblasts before their fusion to osteoclasts. AB - Precursors of osteoclasts seeded on top of a confluent layer of osteoblasts/bone lining cells induced retraction of the latter cells. The (pre)osteoclasts then migrated in the formed cell-free areas and fused to form osteoclast-like cells. Retraction of the osteoblasts/bone lining cells proved to depend on activity of matrix metalloproteinases, and TGF-beta1 prevented the retraction. INTRODUCTION: It is well known that osteoblasts have a profound effect on (pre)osteoclasts in inducing the formation of bone-resorbing osteoclasts. Whether, on the other hand, (pre)osteoclasts also modulate osteoblast activity is largely unknown. Because osteoblasts/bone lining cells have to retract from the surface before resorption of bone by osteoclasts, we addressed the question of whether (pre)osteoclasts have the capacity to induce such an activity. MATERIALS AND METHODS: Rabbit calvarial osteoblasts/bone lining cells or periosteal fibroblasts were cultured until confluency, after which rabbit peripheral blood mononuclear cells (PBMCs) were seeded on top of them. The co-cultures were maintained for up to 15 days in the presence or absence of the cytokines transforming growth factor (TGF)-beta1 and TNF-alpha and selective inhibitors of matrix metalloproteinases and serine proteinases. The formation of cell-free areas and the number of TRACP+ multinucleated osteoclast-like cells were analyzed. In addition, formation of cell-free areas was analyzed in co-cultures of osteoblasts with mature osteoclasts. RESULTS: The seeding of PBMCs on a confluent layer of osteoblasts/bone lining cells resulted in the following sequence of events. (1) A low number of PBMCs strongly attached to osteoblasts. 2) At these sites of contact, the osteoblasts retracted, thus forming cell-free areas. (3) The PBMCs invaded these areas and attached to the surface of the well, after which they fused and formed multinucleated TRACP+ osteoclast-like cells. Retraction was only seen if the cells were in direct contact; conditioned media from cultured PBMCs added to osteoblasts had no effect. Mature osteoclasts seeded on osteoblasts similarly induced retraction, but this retraction occurred at a much faster rate (within 2 days) than the retraction effectuated by the osteoclast precursors (after 8 days in co-culture). Inhibition of matrix metalloproteinase activity, but not of serine proteinases, strongly reduced retraction of the osteoblasts, thus indicating that this type of cell movement depends on the activity of matrix metalloproteinases. A similar inhibitory effect was found with TGF-beta1. TNF alpha had no effect on osteoblast retraction but enhanced the formation of multinucleated osteoclast-like cells. Addition of PBMCs to confluent layers of periosteal fibroblasts resulted in similar phenomena as observed in co-cultures with osteoblasts. However, the cell-free areas proved to be significantly smaller, and the number of multinucleated cells formed within cell-free areas was three to four times lower. CONCLUSION: Our results indicate that osteoclast precursors and mature osteoclasts have the capacity to modulate the activity of osteoblasts and that, yet unknown, membrane-bound signaling molecules are essential in inducing retraction of osteoblasts and the subsequent formation of cell-free areas. PMID- 15355569 TI - Comparison of TGF-beta/BMP pathways signaled by demineralized bone powder and BMP 2 in human dermal fibroblasts. AB - Demineralized bone induces chondrogenic differentiation of human dermal fibroblasts in vitro. Analyses of signaling gene expression showed that DBP and BMP-2 regulate common and distinct pathways. Although BMP-2 was originally isolated as a putative active factor in DBP, rhBMP-2 and DBP do not affect all the same genes or in the same ways. INTRODUCTION: Demineralized bone powder (DBP) induces chondrogenic differentiation of human dermal fibroblasts (hDFs) in 3D culture, but the initiating mechanisms have not been identified. We tested the hypotheses that DBP would affect expression of signaling genes and that DBP's effects would differ from the effects of bone morphogenetic proteins (BMPs). MATERIALS AND METHODS: A chondroinduction model was used in which hDFs were cultured with and without DBP in a porous collagen sponge. BMP-2 was delivered in a square of absorbable collagen felt inserted into a collagen sponge. Total RNA was isolated after 3 days of culture, a time that precedes expression of the chondrocyte phenotype. Gene expression was evaluated with two targeted macroarray screens. Effects of DBP and rhBMP-2 were compared by macroarray, RT-PCR, and Northern hybridization analysis of selected genes in the transforming growth factor (TGF)-beta/BMP signaling pathways. RESULTS: By macroarray analysis of 16 signal transduction pathways, the following pathways were modulated in hDFs by DBP: TGF-beta, insulin/LDL, hedgehog, PI3 kinase/AKT, NF-kappaB, androgen, retinoic acid, and NFAT. There was convergence and divergence in DBP and rhBMP-2 regulation of genes in the TGF-beta/BMP signaling pathway. Smad target genes were the predominant group of DBP- or rhBMP-2-regulated genes. Several genes (IGF-BP3, ID2, and ID3) showed similar responses (increased expression) to DBP and rhBMP-2. In contrast, many of the genes that were greatly upregulated by DBP (TGFBI/betaig h3, Col3A1, TIMP1, p21/Waf1/Cip1) were barely affected by rhBMP-2. CONCLUSION: These findings indicate that multiple signaling pathways are regulated in fibroblasts by DBP, that one of the major pathways involves Smad target genes, and that DBP and rhBMP-2 elicit different gene expression responses in hDFs. Although BMP-2 was originally isolated as a putative inductive factor in DBP, rhBMP-2 and DBP do not affect all the same genes or in the same ways. PMID- 15355570 TI - Maternal and fetal outcome after long-term pamidronate treatment before conception: a report of two cases. AB - The pregnancies of two women with osteogenesis imperfecta who received intravenous pamidronate before conception are reported. The mothers suffered no ill effects. One baby had transient asymptomatic hypocalcemia and one had bilateral talipes equinovarus. This report documents the pregnancy outcomes of two women with osteogenesis imperfecta (OI), types I and IV, who received intravenous pamidronate as part of an observational trial before conception. Pamidronate was not administered after conception. Other than hyperemesis in one woman, the pregnancies and deliveries were uneventful. Both babies inherited OI from their mothers. The baby with OI type IV also had bilateral talipes equinovarus. Biochemical evaluation of the mothers and babies at 24 h and/or 2 weeks postpartum was normal, apart from one baby with asymptomatic hypocalcemia at 24 h of age that had resolved when next measured on day 11 of life. No biochemistry was available on the second child until 13 days of age. Neither baby had skeletal modeling abnormalities consistent with in utero pamidronate exposure. The lumbar spine (L1-L4) areal BMD and anterior to posterior height ratios of lumbar vertebral bodies of both women remained constant during pregnancy. Both the mothers and babies remain well and free of fracture 14 and 16 months postpartum. PMID- 15355572 TI - Development of a reversed-phase thin-layer chromatographic method for artemisinin and its derivatives. AB - In this study a clear separation between seven analogues of artemisinin on thin layer chromatography (TLC) is presented. The developed TLC method is carried out on a RP-C18 thin-layer plate using acetonitrile-water (50:25 v/v) as the mobile phase. Spots are visualized by derivatization with an acidified 4 methoxybenzaldehyde reagent in methanol-water. This method allows the separation of a diverse group of compounds that have versatile hydrophilic/lipophilic characteristics; namely artemisinin, artesunate (AS), artelinic acid (AL), arteether (AE), both isomers of artemether (AM) (alpha and beta), dihydroartemisinin, and desoxyartemisinin. Separation of some degradation products and impurities, down to 2%, allows quality control and stability investigation of all actives in raw material and pharmaceutical formulations. The method is further developed via densitometric measurement for quantitative determination purposes for AL and AS. The derivatization technique is evaluated, showing good stability and reproducibility of the coloring process. Percent relative standard deviation values are less than 5% for replicates, and linearity is obtained in the range of 0.5 to 8 microg. A comparative study with high performance liquid chromatography (HPLC) on a C18 column, applying the same mobile phase, proves the suitability of the TLC method, in which almost all presented analytes are separated from each other. In contrast, HPLC requires at least a 20-min analysis to chromatograph all of the compounds and only betaAM and AE are clearly separated from each other and from the other compounds. PMID- 15355573 TI - Liquid chromatography-accurate radioisotope counting and microplate scintillation counter technologies in drug metabolism studies. AB - The present study involves an analysis of the performance of liquid chromatography (LC)-accurate radioisotope counting (ARC) and microplate scintillation counter (TopCount) technologies in drug metabolism studies. For the purpose of evaluating these systems, biological samples resulting from the metabolism of a radiolabeled [14C] compound, known as compound B, are analyzed using LC-ARC and TopCount under similar high-performance LC conditions. Counting efficiency is 83% for LC-ARC, 77% for TopCount, and linearity is R2 of 0.9998 versus 0.9984, respectively. The limit of detection for LC-ARC is 12 disintegrations per minute (dpm) with 1-min/fraction counting, yet for TopCount it is 8.7 dpm with 5-min/fraction counting. Under optimal conditions for each, the total run time of LC-ARC is approximately half that of TopCount. These results indicate that there is no significant difference between these two systems in terms of efficiency, linearity, and limit of detection. However, the LC-ARC system does not involve any manual operations, yet TopCount requires manual sample transfer and data import. This study shows that impressive progress has been made in the technology of radioisotope counting in drug metabolism using LC-ARC. This system enhances the resolution of radiochromatograms and is able to measure volatile metabolites that TopCount cannot detect at all. The ability to acquire mass spectra online is also a major advancement. The overall results suggest that the combination of LC-ARC with radioactivity detection and mass spectrometry has great potential as a powerful tool for radioisotope measurement in metabolite identification studies during drug discovery and development. PMID- 15355574 TI - Computational chemical analysis of the retention of acidic drugs on a pentyl bonded silica gel in reversed-phase liquid chromatography. AB - A fast method to obtain a quantitative structure-retention relationship is required in chromatography for the rapid optimization of chromatographic separation conditions. Chromatographic data of acidic drugs are analyzed by a computational chemical method to simulate chromatographic simulation. The direct interaction between a model phase and a drug is calculated as an energy value using the molecular mechanics calculation of CAChe. Computational chemistry using a model adsorbent is a new method for quantitative analysis of retention in reversed-phase liquid chromatography. The correlation coefficient is 0.878 (n = 19) between the retention factors of acidic drugs and interaction energy values of the final structure (DeltaFS) between an acidic drug and model pentyl-bonded phase. PMID- 15355575 TI - Determination of the stabilizer sucrose in a plasma-derived antithrombin process solution by hydrophilic interaction chromatography with evaporative light scattering detection. AB - Hydrophilic interaction chromatography (HILIC) is used for the quantitation of sucrose in the range of 10-100 micro g/mL. A poly-2-hydroxyethylaspartamide column is eluted with 25% water-75% acetonitrile, and evaporative light scattering is utilized for detection. A process sample of antithrombin (Atenativ) from Octapharma AB (Stockholm, Sweden) containing 20% sucrose is analyzed. The precision for this high-performance liquid chromatographic method is a percent relative standard deviation (%RSD) of 4, limit of detection (s/n=3) of 1 microg/mL, and mean recovery of spiked samples of 101% (RSD% of 3, n=6). Analysis time is 10 min/sample. Glucose, fructose, sodium citrate, sodium phosphate, Triton X-100, and tri-n-butyl phosphate do not interfere with the method. PMID- 15355576 TI - Liquid chromatographic determination of novel aminithiazolecarboxamide fungicide residues in soil and crops using online solid-phase extraction. AB - A simple and convenient liquid chromatographic method has been developed and applied to the analysis of the novel aminothiazolecarboxamide fungicide, ethaboxam, in soil and crops. After the isolation and concentration of analyte from soil and crops, clean-up and separation of sample solutions are performed by high-performance liquid chromatography with online solid-phase extraction. Good linearity (r2 > 0.9995), recovery [for soil, 95.3-98.4%, and crops (grape, red pepper), 92.9-95.9%], and repeatability are achieved in the calibration range of 0.1-10.3 microg/mL. The limit of detection is the 2.5 parts per billion (ppb) (40 g of soil) and the 20 ppb (25 g of crops), respectively. This assay method shows the suitability for the residual analysis of ethaboxam in soil and crops. PMID- 15355577 TI - An alternative approach for the estimation of equivalent temperature in gas chromatography. AB - The temperature at which the isothermal retention time (tRiso) is equal to the temperature-programmed retention time (tRTPGC) or the isothermal retention index (Iiso) is equal to the temperature-programmed retention index (ITPGC) is defined as the equivalent temperature (Teq). The Teq of one-, two-, three-, and four-step temperature-programmed gas chromatography (TPGC) of unsaturated fatty acid methyl esters (FAMEs) from Chinese mustard seed oil is calculated. All of the tR values obtained at the Teq (tRTeq) are very close to the tRTPGC. The highest difference for each chromatogram is less than 0.5%. The slight deviation may partly arrive from the difficulty in setting the desired carrier gas flow rate. Also, good agreement among equivalent chain length values determined by different methods is observed, including the graphical method at the Teq. Therefore, it is speculated that the proposed method may facilitate the GC identification of FAMEs as well as other organic compounds in TPGC by using the available isothermal retention index database. PMID- 15355578 TI - High-performance liquid chromatographic determination of xanthohumol in rat plasma, urine, and fecal samples. AB - Xanthohumol (XN) is the major prenylated flavonoid in hop plants and as such a constituent of beer. Pharmacological studies have shown that XN possesses marked antioxidant and antiproliferative effects. In order to study the resorption and metabolism of this compound, reversed-phase high-performance liquid chromatography is used for the determination of XN in rat plasma, urine, and feces. In session one, rats receive either oral or intravenous (iv) administration (20 mg/kg body weight) of XN. In session two, rats receive oral administration of 50, 100, 200, 400, and 500 mg/kg body weight XN for bioavailability studies at various dose levels. Plasma, urine, and feces are collected at varying time points and assayed for their XN content. Plasma levels of XN fell rapidly within 60 min after iv administration; no XN is detected in plasma after oral administration in either session. XN and its metabolites are excreted mainly in feces within 24 h of administration. The method is a reliable tool for performing studies of XN in different biological material. PMID- 15355579 TI - Evaluation of deuterium isotope effects in normal-phase LC-MS-MS separations using a molecular modeling approach. AB - Molecular modeling of stationary phases presents a unique challenge because there is little available experimentally derived structural information. Verified interaction mechanisms at a molecular level with analytes are also rare. Molecular mechanics calculations using the Tripos force field were carried out to qualitatively and quantitatively assess stationary phase interactions. Binding energy values of -15.40, 15.28, -12.53, and -12.34 kcal/mol, respectively, are obtained for olanzapine (OLZ), OLZ-D3, des-methyl olanzapine (DES), and DES-D8 that corresponded to the retention behavior of the four compounds observed using liquid chromatography-mass spectrometry (MS)-MS. The model explains, semiquantitatively, the deuterium isotope effect in the normal-phase chromatographic separation of these compounds. PMID- 15355580 TI - Development and validation of chromatographic methods for the identification and quantitation of organic compounds leached from a laminated polyolefin material. AB - Chromatographic methods for the identification of organic compounds leached from a plastic material used in solution containers in the pharmaceutical industry are described. Based on a set of compounds identified in extracts of a multilayered polyolefin film, targeted leachables are delineated for accumulation assessments, and methods to perform target quantitation are developed and validated. PMID- 15355581 TI - GP views of their management and referral of psychological problems: a qualitative study. AB - The ways that GPs manage and make referral decisions for psychological problems are crucial for the provision of mental health care, although the ways they undertake these tasks are not well understood. A long tradition of psychiatric research in the UK has identified significant shortcomings in detection rates, raising concerns that a large proportion of mental health problems go unrecognized and untreated. Other research has suggested that GPs have a different approach to psychological problems than their mental health colleagues, accounting for some of the particular conditions of primary care, although this approach may differ from the methods used by mental health specialists. The present study explored the views of 10 GPs in one geographic locality about managing and making referral decisions for psychological problems. Grounded theory analysis of interview data allowed for the development of a model made up of five components. This model described the ways that GPs explored psychological problems in the context of containing patients' health problems. Referral decisions were made when GPs felt they had reached the limits of their capabilities for treating a particular problem, taking account of the patient's suitability for psychological therapy and access to psychology services. These decisions were influenced by GPs' views of psychological problems and therapies as well as by their professional interactions with psychologists. The implications of the findings for understanding how GPs work with mental health problems and the ways that psychological services can help are discussed. PMID- 15355582 TI - The credibility of psychodynamic, cognitive and cognitive-behavioural psychotherapy in a randomly selected sample of the general public. AB - PURPOSE: To examine the credibility of psychodynamic, cognitive and cognitive behavioural psychotherapy in the general population. METHODS: A random sample (N = 121) of individuals from the county of Ostergotland in Sweden between 20 and 60 years of age rated written descriptions of the three forms of psychotherapy. RESULTS: A clear majority of respondents rated cognitive and cognitive behavioural therapies as more credible than psychodynamic therapy, although the latter was rated as the most credible form of psychotherapy among those who had previous experience of treatment for psychological distress. CONCLUSIONS: Each form of psychotherapy appears to have its own 'market segment' with a high degree of credibility to those within that segment. PMID- 15355583 TI - Moderating variables in the relationship between social comparison and depression: an evolutionary perspective. AB - This study investigated the relationship between social comparison and depression utilizing an evolutionary framework. It investigated the moderating effects on social comparison of the importance of comparison dimensions to the person, and of the perceived importance of the dimensions to other people. An adaptation of a measure of self-esteem (Self Attributes Questionnaire; Pelham & Swann, 1989) was utilized. This consisted of 10-point Likert scale ratings on 10 individual social comparison dimensions (e.g. intelligence, social skills, sense of humour). Questions were added to explore beliefs regarding the importance of social comparison dimensions. Data were collected from a combined clinical sample and non-clinical sample of 174 people. The findings support the prediction that the relationship between social comparison on a particular dimension and depression is moderated by the perceived importance of that dimension in attracting the interest of others. PMID- 15355584 TI - Developing a rating scale for projected stories. AB - The Six-part Story Method (6PSM) is a projective tool in wide use by dramatherapists in the UK, USA and Israel (Lahad & Ayalon, 1993). In contrast to projective tests used by psychotherapists and psychologists, the 6PSM has never been the subject of any validation or reliability studies. This study reports on the identification of scale items to describe the manifest content of six-part stories. In all, 26 statements with acceptable inter-rater reliability have been identified. These statements were used to rate stories produced by clinicians (N = 24) mainstream community mental health patients (N = 21) and patients with a borderline personality disorder (N = 19). Some features that were expected to be indicators of an author with a BPD diagnosis proved to be as common in stories from other authors. However, a scale of eight items was identified that differentiated well between authors with a BPD diagnosis and others, with adequate test-retest and inter-rater reliability. Concurrent validity was tested against the Structured Clinical Interview for DSM-IV Axis II (SCID-II), the Clinical Outcomes in Routine Evaluation Outcome Measure (CORE-OM) and the Inventory of Interpersonal Problems--Short Form (IIP-32). PMID- 15355585 TI - Becoming a self-therapist: using cognitive-behavioural therapy for recurrent depression and/or dysthymia after completing therapy. AB - OBJECTIVES: To explore the ways in which people use cognitive-behavioural therapy (CBT) for recurrent depression and/or dysthymia after leaving therapy. DESIGN: A qualitative interview was used in this study. METHOD: Semi-structured interviews were carried out with nine people who had completed a course of CBT at least three months previously. The interviews explored their use of CBT techniques or models outside of therapy and their everyday management of depression. RESULTS: Eight of the nine participants reported engaging in some self-therapeutic activity, and identified depression, or the threat of depression, as a continuing presence in their lives. They used a range of techniques, either directly transferred from therapy or modified in some way, and identified a number of changes in the way they reacted to difficult situations or negative emotions. These included enactive responses such as leaving the room, making self-efficacy statements, or remembering what the therapist had said to them. Participants also described situations in which they could not use the things they had learnt in CBT. Finally, a range of factors that influenced the ways in which participants became self-therapists were identified. CONCLUSIONS: A number of implications for clinical practice are described. An understanding of how people modify CBT and use it (or not) in their everyday lives is important to understanding and improving effectiveness. PMID- 15355586 TI - Comparison of self-rated and clinician-rated measures of depressive symptoms: a naturalistic study. AB - In order to assess the concordance between self-rating and clinician's assessment tools of depression, as well as factors involved in the differences between auto and hetero evaluation, 198 depressed in-patients were assessed at admission and at discharge using the Montgomery Asberg Depression Rating Scale (10-item version, MADRS) and the self-rating scale Symptoms CheckList (90-item version, SCL-90). We found that about 18% of patients overestimated and about 15% underestimated their depressive symptomatology (SCL-90 depression subscale) relative to the psychiatrist's assessment. Logistic regression analysis showed that the presence of personality disorders and previous history of psychiatric disorders predicted the overestimating group. Discriminant analysis showed that approximately 75% of participants were correctly classified when previous history of psychiatric disorders, presence of personality disorders and age were entered separately into the equation. PMID- 15355587 TI - The Reasons for Depression Questionnaire (RFD): UK Standardization for clinical and non-clinical populations. AB - Recent research into reason giving for depression has illustrated the importance of client beliefs about the cause of their depression. Reasons given have been found to be associated with level of depression, perceived credibility of treatments and therapy outcome. It has been suggested that giving reasons for depression is a form of rule-governed behaviour and as such can cause the depression to be harder to treat (i.e. the reasons become functionally true for the individual). This study investigates the reliability and validity of the Reasons for Depression Questionnaire (RFD; Addis, Truax, & Jacobson, 1995), a 48 item self-report measure developed to measure explanations for the causes of depression. The study provides preliminary normative data for both clinical (n = 123) and non-clinical (n = 105) UK samples. The data indicate high reliability for all subscales including a further subscale (biological) added since the measure was initially developed. Certain subscales correlate significantly with level of depression and specific aspects of self-esteem. This supports the validity of the measure and suggests that it is measuring a distinct concept rather than significantly overlapping with individuals' general beliefs about themselves. PMID- 15355588 TI - Shame as a social phenomenon: a critical analysis of the concept of dispositional shame. AB - An increased clinical interest in shame has been reflected in the growing number of research studies in this area. However, clinically orientated empirical investigation has mostly been restricted to the investigation of individual differences in dispositional shame. This study reviews recent work on dispositional shame but then argues that the primacy of this construct has been problematic in a number of ways. Most importantly, the notion of shame as a context-free intrapsychic variable has distracted clinical researchers from investigating the management and repair of experiences of shame and shameful identities and has made the social constitution of shame less visible. Several suggestions are made for alternative ways in which susceptibility to shame could be conceptualized, which consider how shame might arise in certain contexts and as a product of particular social encounters. For example, persistent difficulties with shame may relate to the salience of stigmatizing discourses within a particular social context, the roles or subject positions available to an individual, the establishment of a repertoire of context-relevant shame avoidance strategies and the personal meaning of shamefulness. PMID- 15355589 TI - Relational tendencies associated with broad personality dimensions. AB - Out-patients (N = 76) completed measures of interpersonal motives and construals derived from relational models theory and a five-factor model personality questionnaire. A broader range of associations between personality dimensions and relational tendencies was obtained than interpersonal circle theorists propose, with dimensions other than extraversion and agreeableness having distinctive relational correlates. The interpersonal domain of personality may therefore be somewhat broader than many mainstream personality theorists suppose. PMID- 15355590 TI - Parental loss before the age of 16 years: a comparative study of patients with personality disorder and patients with schizophrenia in a high secure hospital's population. AB - There is a considerable body of literature supporting an association between separation from parent in childhood and later personality disorder. This study compares a 10-year cohort of high secure hospital patients who had either a personality disorder or schizophrenia, but with no other significant psychiatric comorbidity. The information source was the Special Hospitals Case Register. The most important finding was a statistically significant difference in the rates of child-parent separation between the two groups. In the personality disorder group, 119/147 (81%) of the patients had been separated from one or both parents before the age of 16 compared to 178/289 (62%) in the schizophrenia group. PMID- 15355593 TI - Translational profiling: the genome-wide measure of the nascent proteome. AB - Translation in eukaryotic cells is both physically and temporally separated from transcription. This provides cells with extended options to alter their proteome: (1) directly, by synchronizing translation with an altering transcriptional profile; (2) by imposing a changed translational control over transcripts already present in the transcriptome; or (3) by a combination of (1) and (2). In this paper, recent findings in the controlled translation of the transcriptome using microarray analyses are reviewed. A guide to the current technologies and data analysis is also provided, and future directions in the study of translational control as the interface between the transcriptome and the proteome are outlined. This survey is focused on the yeast Saccharomyces cerevisiae, but the topics covered have universal relevance to the control of translation in eukaryotic cells. PMID- 15355594 TI - Microarray-based analyses of mRNA decay in the regulation of mammalian gene expression. AB - Mammalian cells coordinately regulate their gene expression programmes to ensure appropriate responses to stimuli. While transcriptional events provide an important level of gene expression regulation, modulation of messenger RNA (mRNA) decay provides an additional critical regulatory step. Much of the current knowledge of regulated mRNA decay comes from investigations of cytokine and other early response genes involved in inflammation and immunity. Recent genome-wide measurements of mRNA decay rates have revealed that a much broader repertoire of cellular transcripts are regulated at the level of mRNA decay, suggesting that mRNA decay plays a major role in regulating cellular function. Coupled with recent advances in understanding how signalling pathways impact on mRNA turnover, the regulation of mRNA decay emerges as an important post-transcriptional mechanism for rapidly regulating gene expression. PMID- 15355595 TI - The roles of cytoplasmic poly(A)-binding proteins in regulating gene expression: a developmental perspective. AB - Poly(A)-binding proteins (PABPs) are central to the regulation of messenger RNA (mRNA) translation and stability; however, the roles and contributions of different PABP family members in controlling gene expression are not yet fully understood. In this paper, the current state of knowledge of the different cytoplasmic PABP proteins and their function in animal cells will be summarised, with particular reference to their roles in development. Possible regulatory mechanisms and potential new roles for these proteins in the control of specific mRNAs are also highlighted. PMID- 15355596 TI - Building protein interaction maps for Down's syndrome. AB - Now that the complete sequences for human chromosome 21 and the orthologous mouse genomic regions are known, reasonably complete, conserved, protein-coding gene catalogues are also available. The central issue now facing Down's syndrome researchers is the correlation of increased expression of specific, normal, chromosome 21 genes with the development of specific deficits in learning and memory. Because of the number of candidate genes involved, the number of alternative splice variants of individual genes and the number of pathways in which these genes function, a pathway analysis approach will be critical to success. Here, three examples, both gene specific and pathway related, that would benefit from pathway analysis are discussed: (1) the potential roles of eight chromosome 21 proteins in RNA processing pathways; (2) the chromosome 21 protein intersectin 1 and its domain composition, alternative splicing, protein interactions and functions; and (3) the interactions of ten chromosome 21 proteins with components of the mitogen-activated protein kinase and the calcineurin signalling pathways. A productive approach to developing gene phenotype correlations in Down's syndrome will make use of known and predicted functions and interactions of chromosome 21 genes to predict pathways that may be perturbed by their increased levels of expression. Investigations may then be targeted in animal models to specific interactions, intermediate steps or end points of such pathways and the downstream - perhaps amplified - consequences of gene dosage directly assessed. Once pathway perturbations have been identified, the potential for rational design of therapeutics becomes practical. PMID- 15355597 TI - Pseudogenes in metazoa: origin and features. AB - The complete genome sequences with their annotations are a considerable resource in biology, particularly in understanding the global structure of the genetic material at the molecular level. The reason why some eukaryotic genomes contain large quantities of apparently unnecessary DNA, namely pseudogenes, while others seem to invest in more efficient thinning processes or are equipped with protection systems against parasitic elements still remains a mystery. Several genome-wide surveys have been undertaken to identify pseudogenes in the completely sequenced genome, bringing to light some differences both in their amount and distribution. Since pseudogenes are important resources in evolutionary and comparative genomics - as 'molecular fossils' - in this paper, a survey on the origins, features, abundance and localisation of the different pseudogenes is reported. As an example of genes producing processed pseudogenes, some experimental data obtained in the authors' laboratories from the study of a nuclear gene coding for the mitochondrial transcription factor A (mtTFA), a key regulator of mitochondrial biogenesis, are also reported. PMID- 15355598 TI - Genome-wide RNAi as a route to gene function in Drosophila. AB - With the sequencing of the human genome and the genomes of most major model organisms completed, the systematic characterisation of gene functions remains a key challenge. During the past few years, RNA interference (RNAi) has become a powerful tool to silence the expression of genes and analyse their loss-of function phenotype when mutant alleles are not available. Genome-wide RNAi screens against all predicted genes have been successfully used to dissect a variety of biological processes in Caenorhabditis elegans. Recently, a genome wide library of double-stranded RNAs, that target every gene in the Drosophila genome and that is suitable for high throughput cell-based assays, was published. In this paper, recent advances will be summarised. Screening strategies and applications as a route to comprehensively characterising gene function will be discussed. PMID- 15355599 TI - Development of high throughput dispersive LC-ion mobility-TOFMS techniques for analysing the human plasma proteome. AB - A technique that combines ion mobility spectrometry (IMS) with reversed-phase liquid chromatography (LC), collision-induced dissociation (CID) and mass spectrometry (MS) has been developed. The approach is described as a high throughput means of analysing complex mixtures of peptides that arise from enzymatic digestion of protein mixtures. In this approach, peptides are separated by LC and, as they elute from the column, they are introduced into the gas phase and ionised by electrospray ionisation. The beam of ions is accumulated in an ion trap and then the concentrated ion packet is injected into a drift tube where the ions are separated again in the gas phase by IMS, a technique that differentiates ions based on their mobilities through a buffer gas. As ions exit the drift tube, they can be subjected to collisional activation to produce fragments prior to being introduced into a mass spectrometer for detection. The IMS separation can be carried out in only a few milliseconds and offers a number of advantages compared with LC-MS alone. An example of a single 21-minute LC-IMS-(CID)-MS analysis of the human plasma proteome reveals approximately 20,000 parent ions and approximately 600,000 fragment ions and evidence for 227 unique protein assignments. PMID- 15355602 TI - Real-coded memetic algorithms with crossover hill-climbing. AB - This paper presents a real-coded memetic algorithm that applies a crossover hill climbing to solutions produced by the genetic operators. On the one hand, the memetic algorithm provides global search (reliability) by means of the promotion of high levels of population diversity. On the other, the crossover hill-climbing exploits the self-adaptive capacity of real-parameter crossover operators with the aim of producing an effective local tuning on the solutions (accuracy). An important aspect of the memetic algorithm proposed is that it adaptively assigns different local search probabilities to individuals. It was observed that the algorithm adjusts the global/local search balance according to the particularities of each problem instance. Experimental results show that, for a wide range of problems, the method we propose here consistently outperforms other real-coded memetic algorithms which appeared in the literature. PMID- 15355603 TI - Advanced fitness landscape analysis and the performance of memetic algorithms. AB - Memetic algorithms (MAs) have demonstrated very effective in combinatorial optimization. This paper offers explanations as to why this is so by investigating the performance of MAs in terms of efficiency and effectiveness. A special class of MAs is used to discuss efficiency and effectiveness for local search and evolutionary meta-search. It is shown that the efficiency of MAs can be increased drastically with the use of domain knowledge. However, effectiveness highly depends on the structure of the problem. As is well-known, identifying this structure is made easier with the notion of fitness landscapes: the local properties of the fitness landscape strongly influence the effectiveness of the local search while the global properties strongly influence the effectiveness of the evolutionary meta-search. This paper also introduces new techniques for analyzing the fitness landscapes of combinatorial problems; these techniques focus on the investigation of random walks in the fitness landscape starting at locally optimal solutions as well as on the escape from the basins of attractions of current local optima. It is shown for NK-landscapes and landscapes of the unconstrained binary quadratic programming problem (BQP) that a random walk to another local optimum can be used to explain the efficiency of recombination in comparison to mutation. Moreover, the paper shows that other aspects like the size of the basins of attractions of local optima are important for the efficiency of MAs and a local search escape analysis is proposed. These simple analysis techniques have several advantages over previously proposed statistical measures and provide valuable insight into the behaviour of MAs on different kinds of landscapes. PMID- 15355604 TI - Effective Memetic Algorithms for VLSI design = Genetic Algorithms + local search + multi-level clustering. AB - Combining global and local search is a strategy used by many successful hybrid optimization approaches. Memetic Algorithms (MAs) are Evolutionary Algorithms (EAs) that apply some sort of local search to further improve the fitness of individuals in the population. Memetic Algorithms have been shown to be very effective in solving many hard combinatorial optimization problems. This paper provides a forum for identifying and exploring the key issues that affect the design and application of Memetic Algorithms. The approach combines a hierarchical design technique, Genetic Algorithms, constructive techniques and advanced local search to solve VLSI circuit layout in the form of circuit partitioning and placement. Results obtained indicate that Memetic Algorithms based on local search, clustering and good initial solutions improve solution quality on average by 35% for the VLSI circuit partitioning problem and 54% for the VLSI standard cell placement problem. PMID- 15355605 TI - Automatic generation of controllers for embodied legged organisms: a Pareto evolutionary multi-objective approach. AB - In this paper, we investigate the use of a self-adaptive Pareto evolutionary multi-objective optimization (EMO) approach for evolving the controllers of virtual embodied organisms. The objective of this paper is to demonstrate the trade-off between quality of solutions and computational cost. We show empirically that evolving controllers using the proposed algorithm incurs significantly less computational cost when compared to a self-adaptive weighted sum EMO algorithm, a self-adaptive single-objective evolutionary algorithm (EA) and a hand-tuned Pareto EMO algorithm. The main contribution of the self-adaptive Pareto EMO approach is its ability to produce sufficiently good controllers with different locomotion capabilities in a single run, thereby reducing the evolutionary computational cost and allowing the designer to explore the space of good solutions simultaneously. Our results also show that self-adaptation was found to be highly beneficial in reducing redundancy when compared against the other algorithms. Moreover, it was also shown that genetic diversity was being maintained naturally by virtue of the system's inherent multi-objectivity. PMID- 15355607 TI - [ACC/AHA/ESC treatment guideline for supraventricular arrhythmia]. PMID- 15355608 TI - [Infection control strategy for severe sepsis and septic shock]. PMID- 15355609 TI - [Current status and prospect of critical care medicine in Western Pacific countries and regions: summary of 13th Western Pacific Congress on Critical Care Medicine]. PMID- 15355610 TI - [Prognostic significance of early acute renal failure in patients with cardiogenic shock after acute myocardial infarction]. AB - OBJECTIVE: To investigate the relationship between early developed acute renal failure and the prognosis of patients with cardiogenic shock after myocardial infarction. METHODS: All patients who were consecutively to the General Hospital of PLA from 1993 to 2003 admitted with the diagnosis of acute myocardial infarction or unstable angina in the state of cardiogenic shock were enrolled. In hospital mortality was compared between patients with and without early (<24 hours after onset of shock) developed acute renal failure. Multivariate logistic regression analysis was performed to assess the impact of acute renal failure on the prognosis of these patients. RESULTS: Fifty-one(30%)of 172 patients developed acute renal failure within 24 hours after the onset of shock. In-hospital mortality in patients with and without acute renal failure were 90% (46/51 cases) and 56% (68/121 cases), respectively. Multivariate logistic regression analysis identified acute renal failure as an independent predictor of mortality (odds ratio=6.5; 95% confidence interval: 2.5-18.0; P<0.001). CONCLUSION: Acute renal failure is common in patients with cardiogenic shock and strongly associated with in-hospital mortality. PMID- 15355611 TI - [Effects of angiotensin-(1-7) on remodeling of myocardial collagen network in pressure-overloaded rats]. AB - OBJECTIVE: To investigate the effects of angiotensin-(1-7) on remodeling of myocardial collagen network in pressure-overloaded rats. METHODS: A rat model of pressure-overloaded heart was induced by constriction of abdominal aorta. Seventy five male Sprague Dawley rats were randomized to sham-operated group, model control group and angiotensin-(1-7) group. The rats of the latter two groups were treated with intravenous infusion of either angiotensin-(1-7) (25 microg x kg(-1) x h(-1)) or saline after operation. Ratio of left ventricular weight to body weight (LVW/BW) and myocardial collagen volume fraction (CVF) were determined at 1 and 4 weeks after operation. And types I and III collagen mRNA expressions were assessed with reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: At 1 week after the operation, types I and III collagen mRNA expressions of the left ventricular myocardium in model group and angiotensin-(1-7) group were increased significantly compared with those in sham-operated group. But those in angiotensin-(1-7) group were significantly lower than that in model group. At 4 weeks after the operation, LVW/BW and CVF were increased significantly in model group and angiotension-(1-7) group compared with those in sham-operated group, but those in angiotension-(1-7) group were significantly lower than that of model group. CONCLUSION: Angiotensin -(1-7) can attenuate remodeling of myocardial collagen network in pressure-overloaded rats. PMID- 15355613 TI - [Changes in tissue plasminogen activator and plasminogen activator inhibitor during administration of nitroglycerine into internal thoracic artery in dogs with acute myocardial ischemia]. AB - OBJECTIVE: To study the changes in tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) in a model of acute myocardial infarction reproduced in the dog with administration of nitroglycerin into internal thoracic artery under pressure. METHODS: Sixty healthy cross-breed dogs were randomly divided into experimental and control group with 30 dogs in each group. The animal model of acute myocardial ischemia was reproduced by ligating the left anterior descending coronary artery. Internal thoracic artery in the experimental group was ligated, and a tube was introduced into the proximal end. Nitroglycerine was infused under pressure into the internal thoracic artery in experimental group. The drug was given intravenously in control group. t-PA and PAI were measured before anterior coronary artery ligation and at 0.5, 2 and 6 hours after coronary artery ligation. RESULTS: The t-PA levels in two groups were increased at 0.5 hour after coronary artery ligation, and gradually declined in control group, while no obvious change was found in experimental group. There was significant difference between experimental group and control group at 6 hours after coronary artery ligation(P<0.05). PAI levels were increased after coronary artery ligation, peaking at 6 and 2 hours after coronary artery ligation in both groups. Significant difference in PAI level was observed between two groups at 6 hours after coronary artery ligation(P<0.05). CONCLUSION: Introduction of nitroglycerine through internal thoracic artery under pressure is effective to accelerate release of t-PA from the endothelium while inhibit secretion of PAI, therefore it is useful to modulate the balance of fibrinolysis. PMID- 15355614 TI - [Protection of renal function during interventional diagnostic or therapeutic procedures in coronary heart disease patients complicated by chronic renal insufficiency]. AB - OBJECTIVE: To evaluate the safety and efficacy of protective measures for renal function during interventional procedures in patients with chronic renal insufficiency. METHODS: Changes in renal function in 90 patients of coronary artery disease with chronic renal insufficiency admitted from January 1994 to July 2002 during intraoperative intervention were analyzed. These patients were divided into three groups: in group A patients underwent coronary angiography (SCA) with conventional peri-intervention treatment; in group B patients underwent SCA and treated with low dose dopamine plus saline hydration peri intervention; in group C patients underwent percutaneous transluminal coronary angioplasty (PTCA) and stenting, and treated with low dose dopamine plus saline hydration during peri-intervention. RESULTS: Coronary intervention was successful in all patients of three groups. Contrast medium-associated nephropathy (CAN) was found in 28 patients after the procedures, and them 11 occurred in group A (36.7%), 5 in group B (16.7%), and 12 in group C (40.0%), respectively. The incidence of CAN was highest in group C, and lowest in group B (P<0.01). All patients recovered and discharged after medical treatment, and no hemodialysis was necessary. Higher incidence of CAN was found in patients with congestive heart failure than patients without heart failure (64.5%, 20/31 cases vs. 13.6%, 8/59 cases; P<0.01), and also higher incidence of CAN was found in diabetic patients than non-diabetic patients (42.6 , 23/54 cases vs. 13.9%, 5/36 cases; P<0.01). CONCLUSION: The patients with coronary artery disease with chronic renal insufficiency are able to tolerate the interventional procedures well with safety and ideal prognosis, if they are properly protected with dopamine and fluid during the procedure. PMID- 15355615 TI - [Clinical study on changes of heme oxygenase-1 expression in patients with acute myocardial infarction]. AB - OBJECTIVE: To investigate changes in heme oxygenase-1(HO-1) in patients with acute myocardial infarction. METHODS: Forty-five patients with acute myocardial infarction and 50 with coronary heart disease (diagnosed by coronary angiography) but without acute myocardial infarction were included in this study, and another 40 patients with normal coronary artery as controls. Levels of HO-1 protein expression in monocyte and lymphocyte isolated from the patients were determined by immunohistochemistry and Western blot. Computer picture analyzing system was also used to measure levels of HO-1 protein expression. RESULTS: HO-1 protein expression was located in the cytoplasm. The levels of HO-1 protein expression in patients with acute myocardial infarction were significantly higher than those without acute myocardial infarction (P<0.01). In addition, low levels of HO-1 protein expression were observed in patients with normal coronary artery. CONCLUSION: There is a higher expression of HO-1 in patients with acute myocardial infarction, and a lower expression in patients with normal coronary artery. PMID- 15355616 TI - [Association between serum matrix metalloproteinase-3 concentration and the promoter 5A/6A polymorphism in patients with coronary heart disease]. AB - OBJECTIVE: To investigate the association between serum matrix metalloproteinase 3(MMP-3) concentrations and the promoter 5A/6A polymorphism in patients with coronary heart disease (CHD). METHODS: The study enrolled 137 CHD patients and 106 control subjects without CHD. The diagnosis of both groups were confirmed by coronary artery angiography. One hundred and thirty-seven CHD patients were divided into acute myocardial infarction (AMI)group, unstable angina pectoris (UAP) group and stable angina (SA) group according to World Health Organization criteria. Serum concentration of MMP-3 was measured by enzyme linked immunoadsorbent assay (ELISA). MMP-3 promoter gene containing the 5A/6A polymorphism was amplified by polymerase chain reaction (PCR). PCR products were digested by Tth111 I and then were separated by electrophoresis on agarose gel. RESULTS: The distribution of MMP-3 genotype was not significantly different between CHD patients and controls, so was it between AMI patients and controls. Serum MMP-3 level was significantly higher in AMI group than controls, UAP group and SA group (56.815+/-38.932)microg/L, (39.149+/-24.155)microg/L, (41.640+/ 29.180)microg/L, (33.336+/-20.755)microg/L; P<0.01, P<0.05, P<0.01). Serum MMP-3 levels were not significantly different among genotypes and among controls, UAP and SA groups. No significantly differences in serum MMP-3 levels were found among patients with different numbers of coronary arteries that were involved in CHD. CONCLUSION: No marked association could be found between 5A/6A polymorphism in MMP-3 gene and risk of CHD and AMI. Higher serum level of MMP-3 is strong associated with AMI, while not with number of coronary arteries that are involved in CHD. These data suggest that MMP-3 is a useful marker for AMI, and it might play an important role in the induction of disruption of atherosclerosis plaque. PMID- 15355618 TI - [Characteristics and therapeutic strategies of nosocomial pneumonia in postoperative patients with acute hypertensive intracerebral hemorrhage]. AB - OBJECTIVE: To investigate the characteristics of postoperative nosocomial pneumonia in patients with acute hypertensive intracerebral hemorrhage after evacuation of hematoma and the strategies of prevention and treatment. METHODS: Retrospective analysis was performed on the clinical data of surgically treated patients with acute hypertensive intracerebral hemorrhage in the Department of Neurosurgery from 2000 to 2003. The patients were divided into a group who developed nosocomial pneumonia to compare with a group who did not. Sputum was cultured B and drug sensitivity of isolated bacteria was performed. Appropriate treatment measures were given according to clinical manifestations. RESULTS: In 112 post operative patients with acute hypertensive intracerebral hemorrhage, nosocomial pneumonia developed in 29 patients (25.89%), and 65.99% of microorganisms isolated from respiratory secretion was Gram-negative bacteria. The predominant bacteria were Pseudomonas aeruginosa and Staphylococcus aureus. The mortality was 34.48% (10/29 cases). The prognosis of patients with nosocomial pneumonia was worse than the controls (7.23%, 6/83 cases, P<0.01). CONCLUSION: The preventive and therapeutic strategies for nosocomial pneumonia in postoperative patients with acute hypertensive intracerebral hemorrhage include reduction of risk-factors, surveillance of pathogens, rational use of antibiotics, respiratory and nutrition support, and so on. PMID- 15355620 TI - [Optimal time window for minimally invasive aspiration and drainage of the hematoma in patients with intracerebral hemorrhage]. AB - OBJECTIVE: To seek the optimal time window for minimally invasive aspiration and drainage of the hematoma in patients with intracerebral hemorrhage. METHODS: One hundred and twenty-six patients with intracerebral hemorrhage were divided into two groups in terms of the operation time window: ultra-early stage group (within 6 hours) and early stage group (6-24 hours). European stroke scale (ESS) score and Bartherl index (BI) were used to evaluate the improvement of neurological function. RESULTS: There were no significant differences between the two groups in ESS score, complications, and past medical history on hospital admission. Also no difference existed in the volume of hematoma in different regions of the brain between the two groups, including putamen thalamus and brain lobes. However, one month after the onset, mean ESS score was 64.76+/-7.12 and the mean BI was 66.39+/-7.23 in the ultra-early stage group (non-survivors were excluded), whereas in the early stage group, they were 59.21+/-8.63 and 61.91+/-8.31 (non survivors were excluded too), respectively (both P<0.01). Three months after the mean BI was 79.46+/-13.29 in the ultra-super early stage group and 72.32+/-11.86 in the early stage group, and the difference in the two groups was significant (P<0.01). Though surgery in the ultra-early stage gave better results than that in early stage, no marked difference in mortality was found between the two groups (P>0.05). CONCLUSION: The aspiration and drainage of the hematoma should be done as early as possible (within 6 hours of the onset) provided that the patients are fit to undergo the surgery. PMID- 15355621 TI - [Analysis of causes of late mortality in liver transplant recipients]. AB - OBJECTIVE: To evaluate the cause of late mortality in liver transplant recipients, and to study the prevention and management of these complications. METHODS: Two thousand nine hundred and forty patients who underwent liver transplantation at University of Pittsburgh and survived more than 1 year between February 1981 and April 1998 were included in this study. Cause of death, and other relative factors were analyzed. RESULTS: The rate of death dramatically decreased from 20.4% to 1.9% during that 10 years (1981-1991). Eight hundred and seventeen patients who survived more than 1 year had died during the follow-up period. The causes of death were malignant tumor (20.7%), cardiovascular complication (11.4%), infection (11.3%), and respiratory system complications (9.4%). Following the change in the immunosuppression protocol (1981-1986 was cyclosporin-A, 1986-1990 was OKT3, 1991-1998 was FK506), the 1-year death rate, 5 year death rate, and 10-year death rate were constantly decreased postoperatively. Patients of 3-18-year old group had the lowest mortality rate, while that of 19-60-year old group had the highest mortality rate. The patients who received liver transplantation because hepatic carcinoma or fulminant hepatic failure had the highest mortality after the operation. Mortality of the patients who received liver transplantation because of alcoholic cirrhosis increased gradually year by year. CONCLUSION: Malignant tumor, cardiovascular complication, infection and respiratory system complications are the most common causes of late mortality after liver transplantation. Early diagnosis and treatment can improve the survival rate. PMID- 15355623 TI - [Clinical study on the early nutrition support in postoperative patients with critical hypertensive intracerebral hemorrhage]. AB - OBJECTIVE: To investigate the clinical significance of early nutritional support for patients with critical hypertensive intracerebral hemorrhage (HICH) in a critical condition after operation, and the rationale of different methods of nutritional support. METHODS: One hundred and seven HICH patients after operation with Glasgow coma score (GCS) 6-8 were randomly divided into three groups: early enteral nutrition (EEN) group (38 cases); early parenteral nutrition (EPN) group (35 cases), conventional treatment controls (34 cases). They were given different nutritional supports 48 hours after operation. Changes in nutritional parameters and the clinical complications in three groups after treatment were observed, and the treatment effects after 3 months were compared. RESULTS: At the end of first week after operation, nutrition parameters including albumin (ALb), hemoglobin (Hgb) in EEN and EPN groups were better than those in control group(P<0.05); at the end of second week, they were differences among three groups but without statistical significance (P>0.05). The complications were higher in EPN and control groups (P<0.05 or P<0.01). The outcome was assessed 3 months after the operation in term of activity of daily life (ADL), and the result was better in EEN group than that in EPN and control groups (P<0.005). CONCLUSION: Postoperative HICH patients in critical condition could be benefited with EEN, and complications could be reduced with improved prognosis. PMID- 15355639 TI - [Correlation of expression of RhoA (RhoC and their effector ROCK-1 with malignant phenotype of ovarian cancer cells in vitro]. AB - OBJECTIVE: To investigate the expression of RhoA, RhoC and their effector ROCK-1 in four ovarian cancer cell lines in vitro and their correlation with invasiveness. METHODS: Expression of RhoA, RhoC and ROCK-1 mRNA and protein in four ovarian cancer cell lines SW626, Skov-3, A2780 and Caov-3 was detected by RT PCR and Western blot assay. Invasion assay was done in Boyden chamber. RESULTS: The expression levels of RhoA, RhoC and ROCK-1 mRNA and protein varied in the four different cell lines examined. The expression level of RhoC, but not RhoA and ROCK-1, was significantly correlated with the invasive capability of these cells in vitro (r = 0.95, P < 0.01). Expression of RhoA at the level of transcription was not correlated with that at the translation level. The expression of RhoA and RhoC did not correlate with that of ROCK-1. CONCLUSION: Expression level of RhoC may serve as an independent parameter in evaluating metastasis and become a new target in inhibiting ovarian cancer metastasis. PMID- 15355640 TI - [Altered expression of PTEN gene and LOH of its epigenetic microsatellite in gastric carcinoma]. AB - OBJECTIVE: To investigate the expression of PTEN and loss of heterozygosity (LOH) of its epigenetic microsatellite in gastric carcinoma and explore their roles in progression of gastric carcinoma. METHODS: LOH of epigenetic microsatellites of PTEN (D10S541, D10S583 and D10S1687) in advanced gastric cancer was detected by PCR-SSCP. Expression of PTEN mRNA and protein in normal gastric mucosa and gastric cancer was evaluated by RT-PCR and SABC immunohistochemistry, respectively. The relationship between expression of PTEN mRNA and protein and lymph node metastasis or LOH of microsatellites was discussed. RESULTS: LOH of D10S541, D10S583 and D10S1687 was found in 37.5% (21/56) of advanced gastric cancers. The positive rates of PTEN mRNA expression were 80.4% (45/56), 45.5% (5/11) and 32.1% (18/56) in normal mucosa, early and advanced gastric carcinomas, respectively, while 78.6% (44/56), 44.5% (5/11) and 28.6% (16/56) at the protein level. PTEN mRNA and protein were less frequently expressed in early and advanced gastric carcinomas than that in normal gastric mucosa (P < 0.05). There was positive correlation between PTEN mRNA expression and LOH of microsatellites in advanced gastric carcinomas. PTEN protein expression paralleled with its mRNA expression (P < 0.05). The expression of PTEN mRNA and protein was negatively correlated with lymph node metastasis of advanced gastric carcinomas (P < 0.05). CONCLUSION: Down-regulated expression of PTEN gene is found in different stages of gastric carcinoma, and is closely correlated with LOH of its epigenetic microsatellites, which probably is its underlying molecular mechanisms. It suggests that altered PTEN gene contributes to tumorigenesis and progression of gastric carcinomas. PMID- 15355641 TI - [Epigallocatechin-3-gallate interferes with EBV-encoding AP-1 signal transduction pathway]. AB - OBJECTIVE: To elucidate the interference effect of Epigallocatechin-3-Gallate (EGCG) on targets of Activator Protein-1 (AP-1) signal transduction pathway activated by EB virus encoded latent membrane protein 1 in nasopharyngeal carcinoma (NPC) cell lines. METHODS: Survival rate of cells was determined by MTT assay. AP-1 and CyclinD1 activation were analyzed by promoter luciferase reporter system. Nuclear translocation of JNK was analyzed by indirect immunofluorescence. Protein expression and phosphorylation were observed by Western blot. RESULTS: EGCG inhibited the survival of CNE1 and CNE-LMP1 cells and the activity of AP-1 caused by LMP1 in CNE-LMP1 cells. EGCG also inhibited the nuclear translocation of JNK and the phosphorylation of c-Jun. It also inhibited cyclinD1 promoter activity and cyclinD1 expression. CONCLUSION: EGCG inhibits AP-1, JNK, c-Jun and cyclinD1 which are key targets on AP-1 signal transduction pathway. The results may explain the molecular mechanism of action of EGCG against nasopharyngeal carcinoma. PMID- 15355642 TI - [The relationship between p120ctn translocation and malignant features of hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the effect of catenin p120 (p120ctn) translocation on the malignant features of hepatocellular carcinoma and its interrelation with beta-catenin in E-cadherin-mediated cell signaling. METHODS: Expression and translocation of p120ctn, tyrosine phosphorylation, and its binding capacity to E cadherin were detected by DNA transfection, immunoblotting and immunoprecipitation. Cellular localization of p120ctn and beta-catenin was detected by immunofluorescent microscopy. Cell adhesion, cell migration and cell proliferation were also studied. RESULTS: Expression of p120ctn increased after cells transfected with p120ctn isoform 3A, and it was located mainly at cell-cell contact region. Its binding to E-cadherin was enhanced. After EGF stimulation, tyrosine phosphorylation of p120ctn was increased, membrane expression of p120ctn and beta-catenin was decreased while cytosol expression was increased. It was translocated into the nucleus, cell adhesiveness was increased but mobility decreased. With over-expression of p120ctn, beta-catenin was recruited by nucleus export. Cell proliferation was reduced but it was increased after EGF treatment. CONCLUSION: p120tn plays an important role in cell adhesion, migration and proliferation of hepatocellular carcinoma, and its tyrosine phosphorylation might contribute to this mechanism. There might be a competitive relationship between p120ctn and beta-catenin. PMID- 15355643 TI - [Expression of a plant associated human cancer antigen in breast cancer]. AB - OBJECTIVE: To study the expression of a glycoprotein of plant origin in normal, benign and malignant breast tissues. METHODS: Expression of a plant glycoprotein was examined in 5 samples of normal breast tissues, 20 fibro-adenoma and 136 breast cancer by SABC immunohistochemical staining and the results were analyzed by SPSS statistics software. RESULTS: No positive staining was detected in normal breast tissues (0/5). Weak staining was observed in 4 of 20 (20.0%) breast fibro adenoma. Positive staining was demonstrated in 116 out of 136 (85.3%) breast cancer specimens. The differences were statistically significant. The expression of plant-associated human cancer antigen was related to pathological grade (P < 0.05), tissue invasiveness (P < 0.01) and recurrence (P < 0.05), but not to patients' age, tumor size and c-erbB-2 expression. CONCLUSION: The plant glycoprotein studied may be a human cancer-associated antigen which might be a potential marker of breast cancer. PMID- 15355644 TI - [Endocervical curretage: an analysis of results in 1997 women]. AB - OBJECTIVE: To study the value of endocervical curettage (ECC) in the detection of cervical intraepithelial neoplasia (CIN) and cervical cancer, and the necessity of ECC during colposcopic examination. METHODS: In the high prevalent area of cervical cancer in Shanxi Province, China, a total of 1997 women aged 35 approximately 45 years old were enrolled. Cervical cytology, colposcopy and targeted multiple biopsies, as well as ECC were performed for all women. RESULTS: Among the 1997 women received ECC, 31 was positive for abnormal histologic changes with a frequency of 1.6%. Of the 31 cases, 9 had low grade squamous intraepithelial lesions (LSIL, 0.5%), 20 had high grade squamous intraepithelial lesions (HSIL, 1.0%), and 2 had squamous-cell carcinoma. No pathologic diagnosis could be made in 131 women because the tissue curretaged was insufficient. The women with positive cytologic findings had higher frequency (5.3%) of abnormal ECC than those with negative cytologic findings (0.3%). There was positive correlation between the frequency of abnormal ECC and the grade of cytolologic findings. Abnormal ECC was present in 9.1% of those with unsatisfactory colposcopy while 1.3% of those with satisfactory colposcopy (P < 0.01). The frequency of abnormal ECC was 0.6% in patients with negative colpocopy, 0.9% in LSIL and 24.1% in HSIL. Frequency of abnormal ECC in women with a negative colposcopy or LSIL was significantly lower than that with HSIL. The positive rate of ECC pathologically verified was 3.3% in LSIL, 22.2% in HSIL and 50.0% in squamous carcinoma, respectively (P < 0.01). Of the 316 patients with positive cytology but negative colposcopy, ECC was abnormal in 8 (2.5%), of which HSIL cytologically verified was in 3. CONCLUSION: If cytology or colposcopy shows HSIL or more severe changes, and cytology is positive while colposcopy is unsatisfactory, ECC should be done routinely. PMID- 15355645 TI - [Experimental study of sensitivity to chemotherapeutic agents for human endometrial carcinoma cell line]. AB - OBJECTIVE: To screen the sensitive chemotherapeutic agents to human endometrial carcinoma cell line-1 (HECCL-1) and study its mechanism. METHODS: MTT method was used to examine the relative inhibition ratios (RIRs) of various concentrations of 18 chemotherapeutic agents to HECCL-1. Cell cycle, apoptosis and expression of MDR1 protein were detected by FCM. RESULTS: Nine of the chemotherapeutic agents studied obviously inhibited the proliferative activity of HECCL-1 in a dose dependent manner. The order of sensitivity was as follows: adriamycin (ADM), oxaliplatin (L-OHP), carboplatin (CBP), cisplatin (DDP), taxol (TAL), epirubicin (EPI), mitoxantrone (MIT), dactomycin (ACTD) and 5-fluorouracil (5-Fu). FCM showed these agents could significantly reduce the proportion of cells in G0-G1 phase, and increase the proportion of cells in S and G2-M phase (P < 0.05). Cell apoptosis was observed in 11 chemotherapeutic agents at their peak concentration. MDR expression was induced after using EPI, 5-Fu, hydroxycamptothecin (HCPT) and MIT. CONCLUSION: HECCL-1 is sensitive to a number of the chemotherapeutic agents studied. Induced apoptosis may be the major mechanism of drug sensitivity, and acquired drug-resistance may be the critical reason against continued administration. PMID- 15355646 TI - [Prognostic factors in patients with small cell lung cancer]. AB - OBJECTIVE: To investigate the prognostic factors of small cell lung cancer (SCLC) and establish a reliable model of clinical prognostic index. METHODS: Kaplan Meier and Cox regression were used to analyze the relationship between survival time and prognostic factors in 60 cases of SCLC. The prognostic factors included clinical and laboratory parameters, serum cytokeratin fragment 19 (CYFRA21-1), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), CA125, interleukin 2 (IL-2) and soluble interleukin-2 receptors (sIL-2R). RESULTS: Kaplan-Meier analysis showed that poor prognosis was in patients with KPS < 80 or extensive disease and unrelated to other clinical parameters such as age, sex and smoking index, and in patients with serum NSE > 30 micro g/L, CEA > 5.0 micro g/L, CA125 > 37 KU/L and sIL-2R > 500 KU/L. Serum IL-2 and CYFRA21-1 were also elevated, but had no significant prognostic value. Multivariate analysis indicated that serum NSE, stage and treatment of disease were independent prognostic factors. The three prognostic factors enabled establishment of a prognostic index (PI) based on a simple algorithm: PI = NSE (0 if < or = 30 micro g/L, 1 if > 30 microg/L) + stage (0 = LD, 1 = ED) + CEA (0 if < or = 5.0 microg/L, 1 if > 5.0 microg/L). CONCLUSION: The stage of disease, systemic treatment and the level of serum NSE are independent prognostic factors. Without considering the influence of treatment-related factors on survival, the levels of serum CEA, NSE and stage of disease before treatment are significant independent prognostic factors. PI calculated on the basis of CEA, NSE and stage is recommended to predict the survival of SCLC. PMID- 15355647 TI - [Application of serum protein pattern model in diagnosis of colorectal cancer]. AB - OBJECTIVE: To explore the application of serum protein pattern models in diagnosis of colorectal cancer (CRC) by proteinchip technology. METHODS: One hundred and forty-seven serum samples (55 CRC patients and 92 healthy individuals) randomly divided into training set (n = 87, 32 CRC patients and 55 healthy individuals) and test set (n = 60), were subjected for analysis by surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS). Four top-scored peaks in 5910, 8930, 4476 and 8817 were detected by proteinchip software version 3.0. and were trained by a multi-layer artificial neural network (ANN) with a back propagation algorithm. An artificial neural network classifier had developed for separating CRC from the healthy group. The classifier was then challenged with the test set (60 samples including 23 CRC patients and 37 healthy individuals) to determine the validity and accuracy of the classification system. RESULTS: The artificial neural network classifier separated the CRC from the healthy samples, with sensitivity of 82.6% and specificity of 91.9%. CONCLUSION: Combination of SELDI-TOF-MS with the artificial neural network yields significant higher sensitivity and specificity than CEA in the diagnosis of CRC, which should be further studied. PMID- 15355648 TI - [MR cholangiopancreatography and MR imaging in the diagnosis of extrahepatic cholangiocarcinoma]. AB - OBJECTIVE: To evaluate the imaging features of MR Imaging (MRI) and MR cholangiopancreatography (MRCP) and their clinical value in the diagnosis of extrahepatic cholangiocarcinoma. METHODS: MRI was performed in 54 patients with extrahepatic cholangiocarcinoma proved surgically and pathologically, MRCP in 44 patients, Gadolinium-enhanced in 29 patients. MRI, MRCP and pathological findings were analyzed retrospectively. RESULTS: By MRI, the mass was shown (n = 39) and all bile duct thickened (n = 13) in extrahepatic cholangiocarcinoma. Gadolinium enhanced ones revealed calcified focus (n = 22). By MRCP, interrupted, abruptly cut-off or cone-like changes of the bile duct (n = 16), beak-like or mouse tail changes (n = 26) or tumbler mouth appearance (n = 2) were shown. The bile duct distal to the obstruction was observed in 29 patients. Of the 54 patients examined by MRI in combination with MRCP, correct tumor localization was made in 52 (96.3%) and correct judgement of tumor nature in 50 (92.6%). CONCLUSION: Conventional MRI is an effective supplement to MRCP in the diagnosis of extrahepatic cholangiocarcinoma. MRCP combined with MRI is able to significantly improve the diagnostic accuracy of MR examination. PMID- 15355649 TI - [Clinical features, treatment and prognosis of 136 patients with primary non Hodgkin's lymphoma of the nasopharynx]. AB - OBJECTIVE: To investigate the clinical characteristics, international prognostic index and treatment of primary non-Hodgkin's lymphoma (NHL) of the nasopharynx. METHODS: From January 1983 to December 1997, 136 patients with previously untreated NHL of the nasopharynx were retrospectively reviewed. All patients were confirmed pathologically and classified by Working Formulation system. There were 18 patients with high-grade, 77 intermediate, 2 low-grade and 39 unclassifiable lymphoma. According to Ann Arbor classification, 25 patients had stage I, 91 stage II, 12 stage III and 8 stage IV lesions. Primary therapy was radiotherapy alone in 13 patients and radiotherapy combined with chemotherapy in 12 patients with stage I disease. In 88 patients with stage II, radiotherapy alone was given to 31 patients, and a combination of radiotherapy and chemotherapy to 57 patients. Chemotherapy was primary treatment for advanced stage III/IV diseases. RESULTS: The overall survival rate (OS), cancer specific survival rate (CSS) and disease-free survival rate (DFS) at 5 and 10 years for all patients were 56.2%, 61.2%, 51.1% and 48.3%, 58.0%, 46.5%, respectively. As for international prognostic index (IPI), the 5-year CSS was 70.9% for 0 risk factor, 44.9% for 1 risk factor, 30.0% for 2 or 3 risk factors, respectively (P = 0.004). For stage I patients, the 5-year CSS was 83.1% for RT alone and 82.2% for combined modality therapy, respectively (P = 0.779). For patients with stage II, the 5-year CSS was 46.0% for radiotherapy alone and 70.9% for combined modality therapy. There was significant difference between them (P = 0.04). Multivariate analysis by Cox regression showed that Ann Arbor stage, B symptom and IPI were independent prognostic factors. CONCLUSION: International prognostic index is an important prognostic factor for Non-Hodgkin's lymphoma of the nasopharynx and the combined modality therapy may be optimal for the stage II patients. PMID- 15355650 TI - [Herceptin as a single agent in patients with HER2 overexpressing metastatic breast cancer]. AB - OBJECTIVE: To evaluate the efficacy and adverse reactions of Herceptin as a single agent in patients with HER2 overexpressing metastatic breast cancer. METHODS: There were two kinds of therapy protocol. One was loading-dose of 4 mg/kg intravenously, followed by a 2 mg/kg maintenance dose of weekly intervals. The other was loading-dose of 8 mg/kg, followed by 6 mg/kg of every three weeks intervals. RESULTS: Among 20 patients with evaluated efficacy, there was no complete response, 5 patients (25.0%) showed partial response (PR), 5 (25.0%) stable disease (SD) and 10 (50.0%) progressive disease (PD). Of 22 patients, the overall response rate was 22.7%. The median time of disease progression and treatment failure was 6 weeks and 6.5 weeks, respectively. The most common adverse reactions were fever and chill. Cardiac symptoms could be seen in some patients. CONCLUSION: Herceptin is an active agent for the patients with HER2 overexpressing metastatic breast cancer and the adverse events are well tolerated. PMID- 15355651 TI - [Intraoperative 125I brachytherapy combined with chemotherapy for pancreatic cancer]. AB - OBJECTIVE: To evaluate the efficacy, toxicity and survival of intraoperative 125I brachytherapy combined with chemotherapy for advanced pancreatic cancer. METHODS: Thirty-six patients with advanced pancreatic cancer were randomized to two groups: brachy-chemotherapy group (n = 18) and control group (n = 18). For the combined group, intraoperative 125I implantation and gemcitabine, 5-Fu were given. For the control group, intratumoral injection of absolute alcohol was done. RESULTS: The CR + PR rate of brachy-chemotherapy group was 38.9% with pain relief in 77.8%, while that of control group was 0 with pain relief in 22.2% (P < 0.05). Although there were some toxicity in brachy-chemotherapy group, treatment was well tolerated. The 6-, 12-month survival rates of brachy-chemotherapy group were 71.4% and 21.4% and those of control group were 38.5% and 7.7%, respectively. The median survival time was 10.6 months and 5.2 months for the two groups, between which the difference was significant (P < 0.05). CONCLUSION: Interoperative 125I brachytherapy combined with chemotherapy for advanced pancreatic cancer can control tumor, relieve pain and improve quality of life. It is safe and effective. PMID- 15355652 TI - [Diagnosis and treatment of gastrointestinal stromal tumors: a study of 96 patients]. AB - OBJECTIVE: To investigate the pathological diagnosis, surgical treatment and prognostic factors of gastrointestinal stromal tumors (GISTs). METHODS: The clinicopathological data of operated 96 patients with GISTs were analyzed retrospectively. Expression of CD117, CD34, SMA and S-100 was determined by immunohistochemical methods. RESULTS: Expression of CD117, CD34, SMA and S-100 was 79.2% (76/96), 58.3% (56/96), 35.4% (34/96) and 9.4% (9/96). Benign tumor 23 and malignant 73. Of the malignant, the omentum was resected in 39 and the rest remained, of which the recurrent and metastatic rates were 5.1% and 26.5% (P < 0.05). The safety margin between the normal intestine and tumor was > 5 cm in 46 patients; while in the other 27 patients, it was < 5 cm. The recurrent and metastatic rates were 6.5% and 29.6% (P < 0.05), respectively. The 5-year survival rates of benign and malignant GISTs were 91.5% and 57.3% (P < 0.05). CONCLUSION: The application of immunohistochemical markers CD117 and CD34 are supplementary to pathological diagnosis. The adapting of rational primary treatment, including complete tumor resection and prophylactic omentectomy, is able to reduce the recurrence of GISTs. PMID- 15355653 TI - [Clinical analysis of 38 elderly patients with early double primary cancers]. AB - OBJECTIVE: To study the clinical features and proper treatment of 38 elderly patients with early double primary cancers. METHODS: Thirty-eight elderly patients with early double primary cancers treated from January 1980 to March 2003 were retrospectively reviewed for involved organs, treatment and prognosis. RESULTS: Digestive tract was the most frequently involved, followed by urogenital system and lung. Long-term results of endoscopic mucosal resection (EMR), operation and radiotherapy were superior to other methods. The prognosis of gastrointestinal carcinoma was better than that of prostate carcinoma and hematopoietic system. The operation rate decreased with increasing age. The 5 year survival rates of EMR, operation and radiotherapy were 85.7%, 71.1% and 75.0%, respectively. The medium survival time was 120 months in first cancer and 39 months in the second primary cancer. The 5-year survival rates of the first cancer and second primary cancer were 88.6% and 53.8%. CONCLUSION: Yearly follow up for elderly patients with endoscopy, beta ultrasonic scan and X-ray contribute to finding of early double primary cancers. Operation is the best treatment of early double primary cancers. Endoscopic mucosal resection is especially suitable for old patients with digestive tract and bladder cancer. PMID- 15355654 TI - [A promising attempt in treating systemic lupus erythematosus]. PMID- 15355655 TI - [Evidence-based practice in the management of systemic lupus erythematosus]. PMID- 15355656 TI - [General medicine, the basis of specialty medicine]. PMID- 15355657 TI - [Several suggestions for training internists]. PMID- 15355658 TI - [A pilot study of T cell vaccination in systemic lupus erythematosus]. AB - OBJECTIVE: Autoreactive T cells play a critical role in pathogenesis of systemic lupus erythematosus (SLE). Immunization with inactivated autoreactive T cells (T cell vaccination) may activate the idiotype anti-idiotypic network to deplete specific subsets of autoreactive T cells involved in SLE. We conducted a pilot clinical trial of T cell vaccination to investigate the efficiency and safety of T cell vaccination in treatment of SLE. METHODS: Autoreactive T cell clones were derived from peripheral blood mononuclear cells of 6 SLE patients. After irradiated with 80 Gy gamma radiation, 1 x 10(7) T cells were inoculated subcutaneously at 0, 2, 6, 8 week respectively. The patients were followed up for 20-27 months, and monitored for clinical characteristics and side effects from the vaccination. RESULTS: The clinical manifestations and laboratory abnormalities were improved after inoculation without increasing the dose of corticosteroids and immunosuppressants in most patients. SLEDAI score were decreased remarkably. Proliferative responses against the T cell vaccine were observed in 4/6 patients. No side effect was noticed and CD3+, CD4+ and CD8+ T cell were all in normal ranges after the vaccination and during the follow-up period. CONCLUSION: The results of this pilot study indicate that T cell vaccination is a safe and effective treatment in SLE patients. PMID- 15355659 TI - [The expression of migration inhibitory factor in peripheral blood mononuclear cell and its correlation with disease activity in systemic lupus erythematosus]. AB - OBJECTIVE: To investigate the change of macrophage migration inhibitory factor (MIF) mRNA expression in peripheral blood mononuclear cell (PBMC) and serum MIF and its correlation with disease active index in patients with systemic lupus erythematosus (SLE). METHODS: MIF mRNA expression of PBMC and serum MIF in patients with active and inactive SLE was measured by reverse transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The correlations of serum MIF with SLE disease active index, serum ds-DNA antibody, ANA, C3, C4 and IgG were examined. RESULTS: There was marked increase in MIF mRNA expression of PBMC and serum MIF in patients with SLE than that in normal volunteers (P < 0.01). Level of MIF in patients with active SLE was higher than that with inactive SLE (P < 0.01). There were significant correlation between the blood MIF concentration, MIF mRNA expression and SLE disease active index, ds-DNA antibody, C3, C4 and IgG, but no significant different was observed between the blood MIF and antinuclear antibody. CONCLUSIONS: MIF mRNA expression in PBMC and serum MIF concentration is significantly increased in patients with SLE and correlates with SLE disease activity. Serum MIF levels may be a useful parameter for monitoring disease active in patients with SLE. PMID- 15355660 TI - [Cytophagic histiocytic panniculitis: a report of 6 cases with literature review]. AB - OBJECTIVE: To study the clinical characteristics of cytophagic histiocytic panniculitis (CHP) and increase the understanding of CHP. METHODS: Clinical data of six cases with CHP in our hospital from 1994 were presented with literature review. RESULTS: (1) Six patients including five females, at a mean age of 21.8 year, suffered from the diseases for a mean period of 12.7 months; (2) The major clinical presentations included fever (6/6), subcutaneous nodules (6/6), splenomegaly (5/6) and hepatomegaly (4/6); The laboratory examinations showed elevated hepatic enzymes (4/6), coagulative dysfunction (2/6) and hemocytopenia (3/6); (3) The pathologic examinations revealed benign histiocytes that infiltrated the adipose tissues (6/6) and hemopieotic tissue (3/3), phagocytosing hemocytes; (4) Only one patient was treated with combined chemotherapy. CONCLUSIONS: CHP should be considered based on its unique clinical characteristics and would be diagnosed depending on pathology. The aggressive treatment of combined chemotherapy might improve the prognosis. PMID- 15355661 TI - [Proliferative changes of human gastric mucosa cells in different pathological lesions and their clinical significance]. AB - OBJECTIVE: To investigate the variation of cell proliferation and growth in different pathological lesions of gastric mucosa and to assess the possible roles of expression of epidermal growth factor receptor (EGFR), transforming growth factor beta receptor type I and type II (TGF(beta)RI, TGF(beta)RII). METHODS: The proliferating cell nuclear antigen (PCNA), EGFR, TGF(beta)RIand TGF(beta)RII were studied in chronic superficial gastritis (CSG, n = 30), chronic atrophic gastritis (CAG, n = 26), intestinal metaplasia (IM,n = 40), Dysplasia (DYS, n = 22), early gastric cancer (EGC, n = 22), advanced gastric cancer (AGC, n = 26) by immunohistochemical methods and their relations with carcinogenesis were analyzed. RESULTS: (1) In different gastric mucosa lesions (CSG, CAG, IM, DYS, EGC, AGC), there were significantly different expression of PCNA (chi2 = 91.06, P < 0.0001), EGFR (chi2 = 52.82, P < 0.0001), TGF(beta)RI (chi2 = 15.93, P = 0.007) and TGF(beta)RII (chi2 = 40.48, P < 0.0001), PCNA and EGFR were increased, TGF(beta)RI and TGF(beta)RII were decreased. (2) In DYS stage, PCNALI (40.00 +/- 16.34) was higher than in CSG (16.63 +/- 10.52), CAG (16.92 +/- 8.50) and IM (23.25 +/- 18.64), but lower than EGC (53.09 +/- 13.51) and AGC (57.54 +/- 16.88) (P < 0.0001); (3) EGFR expression in IM (55.0%) and DYS (72.7%) were higher than in CSG (10.0%) and CAG (3.8%) (P < 0.0001), but no different with EGC (59.1%) and AGC (73.1%). (4) TGF(beta)RI expression in EGC (50.0%) and AGC (30.8%) were lower than in CSG (73.3%) (P = 0.007). (5) TGF(beta)RII expression in AGC (26.9%) was lower than in CSG (83.3%), CAG (82.8%), IM (65.0%), DYS (54.5%) and EGC (45.5%) significantly (P < 0.0001). (6) The expression of EGFR had positive correlation with PCNA, TGF(beta)RI and TGF(beta)RII had negative correlation with PCNA respectively, TGF(beta)RI and TGF(beta)RII had positive correlation. CONCLUSIONS: DYS is the key link in the change of cell proliferation during gastric carcinogenesis; The increase of EGFR and the decrease of TGF(beta)R may play important roles in promoting gastric carcinogenesis by affecting gastric cell proliferation. PMID- 15355662 TI - [The predictive value of risk scoring system in myocardial infarction in long term prognosis of patients referred to direct percutaneous coronary intervention]. AB - OBJECTIVE: To evaluate the predicting values of thrombolysis in myocardial infarction (TIMI) risk scoring system for long-term prognosis in patients with ST segment elevation myocardial infarction (STEMI) referred for primary percutaneous coronary intervention (PCI). METHODS: Eight clinical variables and their relative value of score derived from TIMI risk scoring system were used to determine individual's risk score. The patients with STEMI were evaluated during the period of in-hospital and followed-up at a mean of (23.9 +/- 3.8) months for major adverse cardiovascular events (MACE), including nonfatal heart failure, nonfatal re-infarction, target vessel revascularization and cardiac mortality and the predicting value of TIMI risk scores at hospital admission for total MACE were analyzed. RESULTS: Out of 373 patients with STEMI referred for primary PCI, 89 patients were presented with MACE during the period of follow-up (average incidence: 23.9%). The presentation of the total MACE increased progressively with the rising TIMI risk scores (P < 0.05 for trends in increase of MACE). The incidence of total MACE in patients with a score > or = 8 was 9 times those with a score 0. The incidence of cardiac mortality was higher (25% vs 0, P < 0.01) and the combined incidence of cardiac mortality and nonfatal re-infarction also increased significantly (36.7% vs 2.6%, P < 0.01) in patients with a score > or = 6 than in those with a score < 6 respectively. The risk of adverse cardiovascular events increased with rising in TIMI risk scores in patients with either high or normal TnI level. CONCLUSIONS: The higher the TIMI risk score at admission, the worse the prognosis. This TIMI risk scoring system is likely to be a simple and practical tool at the bedside in quantitative risk evaluation and long-term prognosis prediction in patients with STEMI referred for primary PCI. PMID- 15355663 TI - [The clinical features of myasthenia gravis affecting nonskeleton muscles]. AB - OBJECTIVE: To propose a method for diagnosis and differential diagnosis of myasthenia gravis (MG) with heart and liver injury, anisocoria, dysacusis, impatience etc. METHODS: (1) Before and after administering cholinesterase inhibitor and immune therapy, 55 patients with MG were re-examined for heart and liver and followed up. (2) 1 mg neostigmine was injected to the MG patients with anisocoria and dysacusis, the symptoms were observed and brainstem auditory evoked potential was performed before and after 30 minutes and 2 hours. RESULTS: With the relaxation of muscular weakness and fatigability distinctive of MG, other special symptoms will improve and no further special therapy is needed. CONCLUSIONS: MG is an autoimmune disease mediated by auto-antibodies that attack the acetylcholine receptors not only at the skeletal muscle, but also at many other organs such as heart, liver, hearing apparatus pupillary sphincters and genitals. With the relaxation of muscular weakness, these symptoms will improve. The effect of administering cholinesterase inhibitors is the key point of differential diagnosis. PMID- 15355664 TI - [Relationship of plasma homocysteine and folic acid levels and 5,10 methylenetetrahydrofolate reductase gene mutation with venous thromboembolism]. AB - OBJECTIVE: To investigate the association between concentrations of plasma homocysteine and folic acid, 5,10-methylenetetrahydrofolate reductase (MTHFR) C667T mutation and venous thromboembolism (VTE) and to analyze the effect of MTHFR C667T mutation on concentrations of plasma homocysteine and folic acid. METHODS: In 58 patients with VTE and 58 sex and age matched controls, epidemiological risk factors were surveyed. The concentration of plasma homocysteine was measured by high performance liquid chromatography, and the concentration of plasma folic acid was measured by radioimmunoassay. MTHFR C667T genotype was measured by PCR-RFLP. RESULTS: The concentrations of plasma homocysteine and folic acid showed significant difference between the case group and the control group (OR = 1.5, 95% CI: 1.216 - 2.213 and OR = 0.396, 95% CI: 0.149 - 0.709, respectively). There was no significant difference in the frequency of mutant alleles in site 667 of MTHFR gene between the cases and the controls (P > 0.05). The concentration of plasma folic acid was associated with the concentration of plasma homocysteine (multiple correlation coefficient = 2.061, P < 0.05). The MTHFR C667T polymorphism was associated with the concentration of plasma folic acid but not with the concentration of plasma homocysteine in both the case group and the control group. The multiple correlation coefficient between the MTHFR C667T polymorphism and the concentration of plasma folic acid is 0.5856 (P < 0.01). CONCLUSIONS: The results of our study demonstrate that hyperhomocysteinemia and folic acid deficiency are independent risk factors for VTE. Folic acid deficiency is a cause of hyperhomocystinemia while the MTHFR C667T mutation is one of the possible genetic factors causing folic acid deficiency in this Chinese population. PMID- 15355665 TI - [Influence of N-acetylcysteine on the cytokines and matrix metalloproteinases in chronic obstructive pulmonary disease rat models]. AB - OBJECTIVE: To investigate the changes of interferon-gamma (IFNgamma), interleukin (IL)-4, matrix metalloproteinase (MMP)-9, MMP-12 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 in smoke-induced chronic obstructive pulmonary disease (COPD) rat models and the therapeutic effect of N-acetylcysteine (NAC). METHODS: Male Wistar rats were exposed to cigarette smoke for 3.5 months. NAC was given in the last month. Lung function was measured at the end of the study. The levels of IL-4 and IFNgamma in broncho-alveolar lavage fluid (BALF) and lung tissues were determined by ELISA. The expression of MMP-9, MMP-12 and TIMP-1 mRNA in lung tissues were determined by RT-PCR. RESULTS: (1) In comparison with the control group, smoke exposed group presented a significant decrease in forced expiratory volume in 0.3 second (FEV(0.3))/forced vital capacity (FVC), dynamic lung compliance (Cdyn), mean alveolar numbers (MAN) and a significant increase in expiratory resistance (Re), pulmonary mean linear intercept (Lm) (P < 0.05). After treatment with NAC, FEV(0.3)/FVC, Re and Cdyn were improved significantly (P < 0.05). No significant changes were found in Lm and MAN (P > 0.05). (2) In the lung tissues of smoke exposed group, IL-4 level was 10.00 pg/ml, IFNgamma level was 19.37 pg/ml, and the IL-4/IFNgamma ratio was 0.49. In the lung tissues of the control group, they were 4.38 pg/ml, 54.94 pg/ml and 0.10, respectively. There were significant differences in these indexes between the smoke exposed group and the control group (P < 0.05). IL-4 level in the NAC group was 7.99 pg/ml which was similar to that in the smoke exposed group (P > 0.05). IFNgamma and IL-4/IFNgamma ratio were 43.40 pg/ml and 0.15, the former being significantly higher and the latter being significantly lower than those in the smoke exposed group (P < 0.05). (3) The expression of MMP-12 mRNA and MMP-12/TIMP-1 ratio in the smoke exposed group (0.36, 1.21) were significantly higher than those in the control group (0.24, 0.88) (P < 0.05). There was no difference in TIMP-1 (P > 0.05). The expression of MMP-12, TIMP-1 mRNA and the MMP-12/TIMP-1 ratio in NAC group were similar to those in the smoke exposed group (P > 0.05). CONCLUSIONS: (1) Cigarette smoke exposure increased IL-4 and decreased IFNgamma. This may contribute to smoke-induced changes in lung function. NAC had no effect on IL-4, but increased IFNgamma, and the IL-4/IFNgamma ratio returned to normal. This might be one of the mechanisms of NAC in improving lung function. (2) Cigarette smoke promoted MMP-12 gene expression and increased the MMP-12/TIMP-1 ratio. This may play a role in smoke-induced emphysema. NAC did not alter MMP-12/TIMP-1 ratio when given in the late phase of smoke exposure. This result could explain the emphysematous changes in the NAC group. PMID- 15355666 TI - [Clinical analysis of 50 cases of cytomegalovirus disease]. AB - OBJECTIVES: To investigate the clinical features, diagnostic methods, therapeutic approaches, outcomes and the alterations of peripheral lymphocyte subsets in cytomegalovirus (CMV) diseases. METHODS: From August 2000 to October 2002, 50 cases of CMV diseases were treated in Peking Union Medical College Hospital. The diagnosis were made by their symptoms and positive CMV pp65 antigen or/and CMV specific IgM. Of the 50 patients, 15 were male, and 35 female. Their age was (35.5 +/- 16.2) years. Their clinical data were collected, and their peripheral lymphocyte subsets were measured at the time when they were first diagnosed as CMV disease. The lymphocyte subsets of 51 healthy blood donors were tested as normal controls. The lymphocyte subsets were compared between those who had an immunocompromising underlying disease and those who had not. RESULTS: (1) CMV disease occurred in 39 patients who had immunocompromising underlying diseases, and in 11 patients who had not; (2) CMV disease caused various symptoms, most of them were nonspecific. Fever (100%), hematological abnormalities (96%) and abnormal liver function test (30%) were the most common manifestations. 3 cases of CMV retinitis were acquired immune deficiency syndrome patients with CD4+ T cell count less than 50/ microl; 2 patients developed multiple organ failure; (3) The positive rates of CMV pp65 antigen and specific IgM assays were 85.3% and 68.3%, respectively; (4) Comparing with normal controls, all patients with CMV disease had decreased B cell and nature killer cells, increased CD8+ T cell percentage and a reversed CD4+/CD8+ ratio; further more, the patients with underlying diseases had lower CD4+ T-cell and CD8+ T-cell counts than those without underlying diseases. CONCLUSIONS: CMV disease occurred in patients with or without underlying diseases. Their clinical manifestations were heterogeneous and non-specific, therefore, special laboratory tests were necessary to confirm the diagnosis. pp65 antigen assay had more diagnostic value than IgM assay. The dramatic changes in lymphocyte subsets suggest CMV disease has a immunological pathogenesis. PMID- 15355667 TI - [The mechanism of homocysteine in Kkay mice diabetic nephropathy model]. AB - OBJECTIVE: To study the expression of matrix metalloproteinase-9 (MMP-9) in diabetic nephropathy of Kkay mice with hyperhomocysteinemia. METHODS: Sixteen Kkay mice were divided into two groups with 8 in each:diabetes group (KA) and methionine-diet group (KB). C57BL/6 mice were used as normal control (C57). Four months after being treated with two different diets, the mice were sacrificed and serum homocysteine (Hcy) was assayed with fluorescence polarization immunoassay. Renal pathological change was examined with periodic acid Schiff (PAS). The expression of MMP-9 protein and mRNA was detected with immunohistochemical staining and reverse transcriptase-PCR respectively. RESULTS: It was shown that induction of hyperhomocysteinemia in Kkay mice (KB) with a diet enriched in methionine aggravated diabetic nephropathy as compared with the KA group (P < 0.05). The expression of MMP-9 protein and mRNA in KB group was also enhanced and the mean positive area of MMP-9 in KB group was higher than that in KA group (15.90% vs 11.14%, P < 0.05). CONCLUSION: The increased level of Hcy could worsen the nephropathy; this may be related to a higher expression of MMP-9. PMID- 15355668 TI - [A multi-center clinical trial of recombinant human thrombopoietin in chronic refractory idiopathic thrombocytopenic purpura]. AB - OBJECTIVE: To evaluate the efficacy and safety of human recombinant thrombopoietin (rhTPO, a product of Sunshine Pharmaceutical Co Ltd, China) in chronic refractory idiopathic thrombocytopenic purpura. METHODS: Eighty-two patients with chronic refractory idiopathic purpura received daily subcutaneous administration of rhTPO at a dose of 1.0 micro g/kg for a maximum of 14 doses. RESULTS: After the beginning of treatment, the median platelet counts increased from 15.5 (6.0 - 24.0) x 10(9)/L to 27.5 (16.0-47.0) x 10(9)/L, 35.0 (20.5-78.0) x 10(9)/L and 77.0 (41.8-119.5) x 10(9)/L on the fifth, seventh and fifteenth day, respectively (P < 0.01). After the discontinuation of rhTPO administration, the platelet count decreased gradually. On the twenty-eighth day, the median platelet count was 76.5 (35-120.3) x 10(9)/L, which was still significantly higher than the level before the treatment (P < 0.01). The overall response rate was 85.3%. The rate in the group with remarkable response (platelet count > or = 100 x 10(9)/L without bleeding) was 58.5% and that in the good response group (platelet count > or = 50 x 10(9)/L or 30 x 10(9)/L higher than the count before the treatment, without bleeding) was 26.8%. Only 3 patients had mild clinical untoward reactions. Low titer (1:5) of anti-TPO antibody in serum was detected with ELISA in one of the sixteen patients who received the test on the twenty first and twenty-eighth day. Neutralizing test using TPO-dependent cell line showed that the positive serum had no neutralizing activity on rhTPO. CONCLUSION: Consecutive subcutaneous injection of rhTPO for a maximum of 14 days was associated with a temporary elevation in platelet counts in patients with chronic refractory ITP with tolerable adverse effects. PMID- 15355669 TI - [The clinical efficacy of hemoperfusion in treatment of drug poisoning]. AB - OBJECTIVE: To investigate the ability of hemoperfusion to remove some drugs or toxin from the body and its clinical efficacy. METHODS: Sixty-nine cases of poisoning due to tetramine or other drugs poisoning in our hospital between July 1990 and December 2003 were studied. Thirty-four patients among them received conventional treatment (including early gastric lavage, hepatoprotection, diuresis or respiratory support), and the remaining were given conventional treatment and hemoperfusion. Toxin concentration changes in blood before and after hemoperfusion, survival rate, time from come onset to regaining consciousness and convulsion termination and duration of clinical course were compared. RESULTS: In the hemoperfusion group, three patients died, the survival rate was 91.4%, whereas in the non-hemoperfusion group, the survival rate was 85.3% (P > 0.05). Meanwhile the clinical course was markedly shortened in the hemoperfusion group (P < 0.05). The time from coma onset to regaining consciousness and convulsion termination in the hemoperfusion group was significantly shorter than that in the non-hemoperfusion group. The clearance rate was different for different poisons, among them the benzodiazepines had an excellent clearance. CONCLUSIONS: Hemoperfusion could adsorb from blood a different amount of poisons. There is obvious efficacy in shortening clinical course and reducing complications. It could possibly raise survival rate in serious poisoning. PMID- 15355687 TI - [Mutation analysis of SHIP gene in acute leukemia]. AB - OBJECTIVE: The SH2 domain containing inositol 5'-phosphatase (SHIP) is predominately expressed in hematopoietic cells, and is a crucial negative regulator in the development of hematopoietic cells. This paper is to evaluate the role of the SHIP gene in human leukemogenesis. METHODS: Expression of SHIP gene in bone marrow and/or peripheral blood from 32 patients with acute myeloid leukemia (AML), 9 with acute lymphoblastic leukemia (ALL), as well as human hematopoietic cell lines was analyzed by reverse transcription-polymerase chain reaction (RT-PCR), single strand conformational polymorphism (SSCP) and DNA sequencing. RESULTS: RT-PCR showed that all samples expressed SHIP gene. Mutations of SHIP gene were detected in 7 (22%) of 32 AML patients and one (12%) of 9 ALL patients. Interestingly, two missense mutations that had been observed in a AML patient at diagnosis disappeared after complete remission (CR). In addition, in vitro Akt phosphorylation was prolonged and increased following IL-3 stimulation of this patient's cells. CONCLUSION: Our data demonstrate for the first time the mutation of SHIP gene in acute leukemia and suggest a possible role of the mutation of this gene in the development of acute leukemia. SHIP may serve as a tumor suppressor by negatively regulating the PI3K/Akt signaling pathway in hematopoietic cells. PMID- 15355688 TI - [Comparison of autologous and allogeneic hematopoietic stem cell transplantation for 140 patients with de novo acute leukemia in first complete remission]. AB - OBJECTIVE: To evaluate the outcome of patients with de novo acute leukemia (AL, no AML-M(3)) in CR(1) undergone autologous hematopoietic stem cell transplantation (auto-HSCT) or HLA-identical sibling allogeneic HSCT (allo-HSCT). METHODS: Forty-six AL patients received allo-HSCT and 94 received auto-HSCT in CR(1). The conditioning regimens mainly consisted of TBICy, BuCy and MAC. Cyclosporine plus methotrexate, or cyclosporine alone, or FK506 alone was used for graft-versus-host disease (GVHD) prophylaxis. Among auto-HSCT group, 39 patients received purged autologous bone marrow and 38 received immunotherapy and/or maintenance chemotherapy after transplant. RESULTS: Myeloid reconstitution was achieved in all patients. After a median of 700 (range, 18 approximately 5563) days follow-up, the probabilities of leukemia-free survival (LFS) at 5 year were not significantly different in these two groups: (51.5 +/- 5.4)% for auto HSCT group and (52.8 +/- 7.6)% for allo-HSCT group (P > 0.05). There was a lower cumulative relapse incidence (RI) [(26.3 +/- 6.9)% vs. (52.0 +/- 5.5)%, P > 0.05] but a significantly higher cumulative transplant-related mortality (TRM) [(37.6 +/- 7.8% vs. (14.4 +/- 4.1)%, P < 0.05] in the allo-HSCT group than in auto-HSCT group. Among auto-HSCT group, the patients received purged autografts and/or post transplant therapy had significantly better LFS and lower RI (P < 0.05) than those received unpurged autografts or no post-transplant treatments [5-y LFS: (62.8 +/- 6.8)% and (38.4 +/- 8.4)%; RI: (37.7 +/- 6.8)% and (74.2 +/- 8.7)%, respectively]. CONCLUSION: The long-term LFS of auto-HSCT was comparable to that of allo-HSCT in the management of patients with AL in CR(1), because autograft purging and post-transplant treatment can significantly decrease relapse of auto HSCT patients and auto-HSCT has lower therapy-related toxicities. PMID- 15355689 TI - [Prevalence and clinical significance of FLT3 internal tandem duplication mutation in acute leukemia]. AB - OBJECTIVE: To evaluate the prevalence of FLT3 mutation-internal tandem duplication in acute leukemia (AL) patients and its significance. METHODS: Genomic DNAs from 194 cases of AL were screened by polymerase chain reaction (PCR) and gel electrophoresis for FLT3-ITD mutations. RESULTS: FLT3-ITDs were found in 37 (25.9%) of 143 de novo acute myeoloid leukemia (AML) patients, including 10/53 of M(2), 15/40 of M(3), 4/20 of M(4) and 8/23 of M(5). Significantly more FLT3 aberrations were found in AML M(3) and M(5) (P < 0.05). No FLT3-ITD was found in 25 acute lymphoid leukemia (ALL), 2 acute hybrid leukemia, 17 myelodysplastic syndromes and 7 chronic myelogenous leukemia in blast crisis. Sequence analysis of 5 cases with abnormal PCR electrophoretic patterns revealed that the ITDs were located within exon 14 from 21 bp to 60 bp, in 3 cases the ITD was a simple tandem duplication, and in 2 cases the ITD was tandem duplication with insertion, but all of the above ITD did not altered the FLT3 reading frame. FLT3-ITD was associated with a higher peripheral white cell count (P < 0.05) and a lower complete remission rate (P < 0.05), and was more prevalent in patients with normal karyotype (P < 0.05). CONCLUSION: FLT3-ITD mutation occurs with a significant percentage in AML M(3) and M(5) patients. Sequences of the mutants are in frame mutation. FLT3-ITD mutation was associated with a higher peripheral white cell count and a lower complete remission rate. PMID- 15355690 TI - [Cell origin of localized Castleman's disease of hyaline-vascular type]. AB - OBJECTIVE: To investigate the cell features of 6 Castleman's disease patients and evaluate their prognosis. METHODS: The resected tumors were investigated by routine histopathology and immunohistochemistry. Reverse-transcript PCR (RT-PCR) and sequencing of RT-PCR products were used to assess the clonal characters of the main tumor cells. RESULTS: Histologically, all of the 6 tumors were classified as the hyaline vascular type. B-cells dominated the follicular germinal centers, with T-cells dispersing inter-follicularly. The results of RT PCR each obtained a single band of either 128 bp or 122 bp and sequencing showed that there was highly homogeneity within the same length sequences, accompanied by fewer different nucleotide acids. CONCLUSION: Monoclonal and/or oligoclonal B cells were identified in Castleman's disease. These B cells were originated from germinal center cells. PMID- 15355691 TI - [The expression of CD133 in acute leukemia and its clinical significance]. AB - OBJECTIVE: To evaluate the expression of CD133 and its clinical significance in acute leukemia (AL) patients. METHODS: The expression of CD133 and CD133 mRNA in leukemic blasts from 76 AL patients were detected by three-color flow-cytometry and hemi-quantitative RT-PCR respectively. RESULTS: (1) CD133 mRNA expression was highly correlated with CD133 expression in both of normal donors and AL patients groups. The expression of CD133 in AL patients was significantly higher than that in control group (P < 0.01). (2) The positive rates of CD133 and CD133 mRNA in AL group were 42.1% (32/76) and 46.1% (35/76) respectively. There was no significant difference in CD133 expression between AML-M(3) and normal control, AML and ALL, as well as T-ALL and B-ALL. The expression of CD133 in AML-M(4) were significantly higher than those in other AML subtypes (81.8% vs 43.7% and 81.8% vs 46.9% at CD133 and CD133 mRNA level, respectively, P < 0.01). (3) The expression of CD133 in AML was significantly correlated with the expression of CD34 and HLA-DR (P < 0.001). (4) The expression of CD133 had no relationship with the clinical prognostic factors such as cytogenetic or molecular aberrations, WBC counts, LDH, mdr1 expression and age. (5) There was a trend toward lower CR rate in CD133(+) cases, but only CD34/CD133(+) double positive cases had significant lower CR rate than that of negative ones (44.4% vs 71.4%, P < 0.05). CONCLUSIONS: AL had significantly higher CD133 expression compared to normal control. The detection of CD133 expression might help to identify AL type and predict therapeutic outcomes. Co-expression of CD133/CD34 might convey adverse prognosis of AL. PMID- 15355692 TI - [The expression of CXCR4 on acute leukemia cells and its implication for extramedullary infiltration]. AB - OBJECTIVE: To study the expression of CXCR4 in acute leukemic cells and its clinical significance. METHOD: Bone marrow samples from 73 acute leukemia patients and leukemic cell lines were investigated by flow cytometry (FCM), the expression of SDF-1 in human marrow stromal cells and meninges were studied by using reverse transcription polymerase chain reaction (RT-PCR). Adhesion, migration and invasion of U937, NB4 and K562 cells were studied in vitro. RESULTS: The expression rates of CXCR4 in ALL and AML patients was 65.6% and 17.1%, respectively. And it was 0.2%, 41.0% and 52.0% in K562, U937 and NB4 cells, respectively. The extramedullary infiltration rates were 61.9% and 18.2% for CXCR4 positive and negative groups of ALL, respectively (P < 0.05); while in AML, the number of peripheral white blood cells in CXCR4 positive group was lower than that in CXCR4 negative group (P < 0.05). SDF-1alpha could enhance the adhesion, migration and invasion capacity of leukemic cells in vitro. CONCLUSION: Overexpression of CXCR4 in AL cells might be the molecular mechanism of extramedullary infiltration in leukemia. PMID- 15355693 TI - [Two Ph chromosome positive chronic myelogenous leukemia patients with rare bcr/abl fusion gene]. AB - OBJECTIVE: To investigate the unusual bcr/abl fusion gene structures of two Ph chromosome positive chronic myelogenous leukemia (CML) patients in chronic phase (CP). METHODS: By using general M- and micro -bcr/abl specific primers respectively, bcr/abl fusion transcripts were detected by reverse transcription polymerase chain reaction (RT-PCR). The RT-PCR products sequencing was performed, the DNA sequences were analyzed in Genebank and the bcr and abl sequences at the fusion site were identified. DNA was amplified by PCR using a set of primers designed according to the sequencing result of RT-PCR products. RESULTS: Two patients showed typical manifestations of CML-CP. Their RT-PCR products were different from usual M- or micro -type; one was longer than M-bcr/abl but shorter than micro -bcr/abl, the other one was shorter than M-bcr/abl. The RT-PCR products sequencing showed that both products contained bcr and abl gene sequences. The first patient's bcr gene was broken within exon 18, and fused to abl gene exon 2(a2), and a 40 bp of partial abl intron 1b fragment was inserted between them, resulting in a novel in-frame bcr/abl fusion transcript-e18-int-a2 which has not been reported in the literature so far. In the second patient, deletion of abl exon2(a2) led to exon 13(b2) of bcr gene fusing with abl exon 3(a3). CONCLUSION: Uncommon bcr/abl fusion gene may occur in typical Ph(+) CML patient. PMID- 15355694 TI - [A combined assay of multiplex RT-PCR and karyotypic analysis in childhood acute lymphoblastic leukemia]. AB - OBJECTIVE: To study the value of combination assay of multiplex RT-PCR and karyotypic analysis in the diagnosis and classification of childhood acute lymphoblastic leukemia (ALL). METHODS: Fifty cases of childhood ALL patients were studied by multiplex RT-PCR in combination with R or G banding karyotype analysis. RESULTS: Of the 50 childhood ALL patients, 18 (36.0%) carried 11 types of fusion genes including E2A/PBX1, TEL/AML1, TLS/ERG, MLL/AF4, MLL/AF9, MLL/AF10, MLL/AFX, MLL/AF6, MLL/ELL, TAL1D, and HOX11, revealed by multiplex RT PCR, and in 48 cases, 24 (57.1%) had chromosome abnormalities. Among the latter, numeral chromosome abnormalities and chromosome deletions accounted for 75.0% (18/24), while translocations 25.0% (6/24). The multiplex RT-PCR in combination with chromosome analysis could detect genetic abnormalities in 70% (35/50) of childhood ALL. CONCLUSIONS: Multiplex RT-PCR combined with chromosome analysis can enhance the detection rate of genetic abnormalities in childhood ALL. It provides reliable evidence for the diagnosis, classification and prognosis. PMID- 15355695 TI - [Analysis of the factors associated with prognosis in patients with Ph chromosome positive adult acute lymphoblastic leukemia]. AB - OBJECTIVE: To investigate factors associated with survival of patients with Ph chromosome positive adult acute lymphoblastic leukemia (aALL) in a period of 11 years. METHODS: All the clinical parameters of 31 Ph positive patients were statistically analyzed by SPSS software. RESULT: Ph(+) patients account for 15.3% (31/203) of all the aALL patients. Clinically, these patients manifested older in age, higher white blood cell counts with high blast fractions and lower platelet counts (PC). Phenotypically 82.6% of them were common ALL, 39.1% coexpressed myeloid antigens, and 56.5% expressed CD34 antigen. 65.4% of them (17/26) achieved complete remission (CR) and the median remission and survival durations were 4 months and 8 months, respectively. Patients with Ph(+) and additional chromosomal aberrations accounted for 42% of all the Ph(+) patients, including monosomy 7, +Ph, del(9)(p11-12) and add/t(16)(p13), and they had lower PC as compared with those with sole Ph(+) (P = 0.012) and variant Ph translocation (P = 0.01). CD34 positive patients had a shorter remission and survival duration than CD34 negative ones (0 vs 9 months for median remission time, P = 0.024; and 6 vs 12 months for median survival time, P = 0.034). There was no evidence to support the correlation between myeloid antigen expression and survival time in these patients. CONCLUSION: Ph(+) aALL is associated with adverse prognosis and CD34 expression is a poorer prognostic factor in Ph(+) aALL patients. There is no significant clinical difference between Ph(+) aALL with or without additional chromosomal aberrations. PMID- 15355696 TI - [MICM classification and prognostic analysis of 80 adolescents with acute lymphoblastic leukemia]. AB - OBJECTIVES: To explore MICM classification and adverse prognostic factors in adolescents with acute lymphoblastic leukemia (ALL). METHODS: The MICM classification, clinical characteristics of 80 adolescents with ALL admitted to our hospital from January 1998 to December 2002 were retrospectively analyzed. Survival data were estimated by the Kaplan-Meier method and the prognostic factors were analyzed with the COX regression model. RESULTS: In the 80 patients, B-ALL and T-ALL accounted for 69.12% and 26.47%, respectively. The percentage of Ph(+)ALL was 18.37% (9/49), and that of hyperdiploidy was 4.08%. Patients at diagnosis with high leukocyte counts (> 50 x 10(9)/L) accounted for 27.94%. Among the 78 cases treated with VDP(L) or CODP(L) regimens, 73 (91.03%) obtained CR in 4 weeks. After a median follow-up of 24 months, the estimated 3-year disease-free survival (DFS) rates of patients receiving chemotherapy or allo-HSCT were (32.55 +/- 16.50)% and (69.58 +/- 8.72)%, respectively (P < 0.05). In COX analysis, high initial leukocyte counts (> 50 x 10(9)/L) and Philadelphia chromosome positivity were adverse prognostic factors for long-term survival. CONCLUSIONS: MICM classification has important clinical and prognostic significance in the risk directed therapy of adolescents with ALL. The adverse prognostic features for these patients were high leukocyte counts, less incidence of chromosome hyperdiploidy and Ph chromosome positivity. PMID- 15355697 TI - [Establishment of a real time quantitative-PCR assay for detection of TCR VgammaI Jgamma gene rearrangement in acute lymphoblastic leukemia patients]. AB - OBJECTIVE: To improve the techniques for minimal residual disease (MRD) detection in acute lymphoblastic leukemia (ALL). METHODS: A real time quantitative PCR method was established for quantifying the clonal TCRVgammaI-Jgamma gene rearrangement in 36 ALL patients. RESULTS: The sensitivity of the established real time quantitative PCR was at 10(-4) level. The amount of TCRVgammaI-Jgamma gene rearrangement in newly diagnosed group, complete remission (CR) group and post hematopoietic stem cell transplantation (HSCT) group was (7.38 +/- 6.65) x 10(-2), (1.02 +/- 1.08) x 10(-2) and (3.89 +/- 5.65) x 10(-3) level, respectively. and the amount in newly diagnosed group was higher than that in CR group and HSCT group (P = 0.001). The MRD level of ALL patients in CR group was higher than that in HSCT group (P = 0.022). MRD can be detected in 6 ALL patients after HSCT, 2 of them with low MRD level (< 1 x 10(-3)) survived long disease free survival, the other 4 with high MRD level relapsed within one year. CONCLUSION: The established real time quantitative PCR assay is simple, rapid, sensitive and specific. Use of this assay to evaluate MRD in the remission ALL cases is helpful for prognosis prediction. PMID- 15355698 TI - [Potential effect of tumor necrosis factor-alpha and its receptor II gene polymorphisms on the pathogenesis of coal worker's pneumoconiosis]. AB - OBJECTIVE: To approach the effect of tumor necrosis factor-alpha (TNF-alpha) and tumor necrosis factor receptor II (TNFRII) gene polymorphisms on genetic susceptibility of coal worker's pneumoconiosis and their relationship with pulmonary fibrosis. METHODS: Two hundred and thirty-four cases of coal worker's pneumoconiosis (CWP) and four hundred and forty coal mine workers (controls) were selected, and the cases of CWP were divided into three subgroups based on the various stages of I, II and III. 3 ml peripheral vein blood was drawn from every subject. Using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) techniques, TNF-alpha and TNFRII gene polymorphisms were analyzed. RESULTS: In both group matching and 1:1 paired matching, there was no significant difference between CWP workers and controls in distribution frequencies of G/A + A/A (TNF-alpha -308) and T/G + G/G (TNFRII 196) genotypes. The distribution frequency of G/A + A/A genotype in CWP with stage III (20.00%) was higher than those in control (10.91%), and CWP cases with stage I (10.34%) and II (7.50%) respectively. The risk of CWP with stage III in those with G/A + A/A genotype was 2-fold higher than with G/G genotype (OR = 3.00, 95% CI: 0.35 approximately 25.84) for 1:1 paired matching. CONCLUSIONS: TNF-alpha and TNFRII gene polymorphisms does not play an important role in susceptibility to CWP of Han race. TNF-alpha gene promoter polymorphisms might be related with the degree of severe pulmonary fibrosis in CWP. PMID- 15355699 TI - [Effect of genetic polymorphisms of microsomal epoxide hydrolase on urinary 1 hydroxypyrene levels in coke oven workers]. AB - OBJECTIVE: To investigate the associations of polymorphisms of metabolic enzyme genes with urinary 1-hydroxypyrene levels in coke oven workers. METHODS: One hundred and forty-eight workers from a coke oven plant and 69 controls without occupational PAHs exposure were selected in this study. Urinary 1-hydroxypyrene was detected by high performance liquid chromatography with florescence detector. The genotypes at I462V site in exon 7 of CYP1A1 gene, GSTM1, GSTT1, I105V site in GSTP1gene, Pst1 and Dra1 sites in CYP2E1 gene, P187S site in NQO1 gene, Kpn1, BamH1 and Taq1 sites in NAT2 gene, and H113Y, R139H sites in mEH gene were determined by PCR-based methods. Personal information including occupational exposure history, age, sex, smoking and drinking status was collected by the questionnaire. RESULTS: The level of urinary 1-hydroxypyrene in coke oven workers [(5.61 +/- 1.04) mol/mol Cr] was higher than that in control [(0.74 +/- 0.32) micro mol/mol Cr]. After adjusting external occupational exposure category and smoking, coke oven workers with variant homozygotes at H113Y site of mEH gene had significantly higher urinary 1-hydroxypyrene concentrations than those with heterozygotes, and wild homozygotes (6.41 +/- 1.09 vs. 6.24 +/- 1.08, and 4.62 +/ 0.95 micro mol/mol Cr, P < 0.05), and gene-gene interaction was found between CYP1A1 and mEH. CONCLUSION: Genetic polymorphism of mEH gene could be a susceptible biomarker in coke oven workers which was involved in the individual susceptibility on metabolism of PAHs. PMID- 15355700 TI - [Relationship of polycyclic aromatic hydrocarbon exposure with peripheral blood lymphocyte DNA damage in coke oven workers]. AB - OBJECTIVE: To investigate the relationship between lymphocyte DNA damage and polycyclic aromatic hydrocarbons (PAHs) exposure in coke oven workers. METHODS: Two hundred and thirty-five coke oven workers and 30 controls were selected in this study. Alkaline single-cell gel electrophoresis was used to evaluate the lymphocyte DNA damage, HPLC was employed to measure 1-hydroxypyrene levels in spot urine samples which were obtained at the end of a workweek (4 days of 8 hours/day) and personal information including occupational exposure, age, sex, smoking and drinking status was collected by the questionnaire. RESULTS: The lymphocyte DNA damage level expressed as olive moment in coke oven workers was significantly higher than that of controls [2.47 (0.22 approximately 46.68) vs 0.94 (0.42 approximately 4.21), P < 0.01], and correlation between urinary 1 hydroxypyrene concentrations and olive moment was found (Spearman Partial correlation coefficient = 0.22, P < 0.01) in coke oven workers. The 1.9 of olive moment value was used as the limit to determine whether the subject DNA damage was positive. The coke oven workers had significantly higher risk in DNA damage (adjusted OR = 5.38, 95% CI = 2.07 approximately 14.08) than did controls, and dose-response relationships were also found between external exposure (exposure category) or internal doses (urinary 1-hydroxypyrene) and DNA damage. CONCLUSION: There are dose-effect and dose-response relationships between PAHs exposure and lymphocyte DNA damage in coke oven workers. PMID- 15355701 TI - [Effect of lumbar protective belt on prevention of low back fatigue in personnel during simulated driving]. AB - OBJECTIVE: To observe low back fatigue of subjects before and after simulated driving, and to study the role of the belt used in prevention of low back fatigue of drivers during driving. METHODS: To assess the changes in median frequency (MF), mean power frequency (MPF) of surface electromyogram (SEMG), and flicker frequency, visuognosis persistence of drivers with and without protective belt in simulated driving. RESULTS: There were significant differences in MF [(47.35 +/- 6.07), (39.26 +/- 5.79), (47.21 +/- 6.02), (43.44 +/- 6.26) Hz respectively], MPF [(69.86 +/- 7.08), (59.12 +/- 7.19), (69.86 +/- 7.08), (63.88 +/- 7.49) Hz respectively] between before and after simulated driving without or with the special belt (P < 0.01). But no differences in MF or MPF between without and with the belt before simulated driving were found (P > 0.05). The flicker frequency [(35.64 +/- 2.82), (42.31 +/- 4.68), (35.96 +/- 3.05), (39.79 +/- 3.36) Hz], visuognosis persistence (65.77% +/- 3.94%, 56.83% +/- 5.68%, 65.88% +/- 3.92%, 62.27% +/- 2.91%) had some changes too. CONCLUSION: Simulated diving could induce fatigue of back muscle in drivers. However, the protective belt can effectively prevent low back fatigue of drivers in simulated driving. PMID- 15355702 TI - [The tendency of physiological and psychological change of the armored vehicle drivers working continuously in hot environment]. AB - OBJECTIVE: To probe the tendency of physiological and psychological change of armored vehicle drivers working continuously in hot environment. METHODS: Ten cross-designed standard subjects were divided into control group and driver group. The driver group worked continuously in 32 degrees C approximately 38 degrees C and RH 50% approximately 80% environment. Heart rate, body temperature, systolic blood pressure and pulse pressure were synchronously and dynamically determined during work. Psychological parameters such as profile of mood states (POMS) questionnaire, serial addition/subtraction (SAS) and two-digit search (TDS) selected from neurobehavioral evaluation system (NES) before and after work were also tested. RESULTS: The curves of the four main physiological parameters rapidly synchro-rose to a peak at (36 +/- 3) min, whereafter the curves descended smoothly. At (144 +/- 8) min, the curves climbed up to the second peak. The determination of the behavioral psychology revealed that the degree of fatigue of the drivers increased gradually when the working time prolonged, and the grip descended by great extent, excitement reduced markedly after work, but the negative mood, such as confusion and depression were increased, error of visual apperceive-operation was increased, right decreased, intelligence reduced, rapidity and delicacy of movement reduced. CONCLUSION: The physiological parameters of armored vehicle drivers working continuously in the hot condition appeared double-peak effect, the 1st peak was at (36 +/- 3) min, and the body was in mobilizing stage. Whereafter, the body was in smooth stage. And at (144 +/- 8) min, when the 2nd peak appeared, the body then was in physiological tolerance limit stage. PMID- 15355703 TI - [Effect of environment and occupational hygiene factors of hospital infection on SARS outbreak]. AB - OBJECTIVE: To study the effects of weather conditions and occupational hygiene on SARS outbreak. METHOD: (1) Meteorological parameters around SARS outbreaks in 2003 in 9 cities (Guangzhou, Beijing, Tianjin, Taiyuan, Hong Kong, Taipei, Singapore, Toronto and Hanoi) were analyzed; (2) Causes of hospital infection were also analyzed from an occupational hygiene point of view. RESULTS: (1) The amplitude of air temperature, air pressure and diurnal temperature difference were greater around SARS outbreaks in most of the cities. Higher airborne particles concentration and lower wind speed were measured prior to SARS outbreaks in the cities with the most serious epidemic situation. The ten-day mean value of air temperature before SARS outbreaks in 9 cities was 16.6 degrees C +/- 7.6 degrees C, suggesting that coronary virus infection, which has been considered to cause SARS by now, may be most active at 9 degrees C - 24 degrees C. (2) Occupational hygiene in hospital proved to be an important socio-behavior factor for SARS outbreak. All hospital infection could be attributed to defects in the key links of occupational hygiene. CONCLUSIONS: Greater fluctuations of air temperature and higher airborne particles concentration in winter and spring, as well as poor occupational hygiene conditions are significant promoters of SARS outbreak. Warning of atmospheric conditions favorable to SARS, and improvement in occupational hygiene management is the key to prevention from SARS outbreak. PMID- 15355704 TI - [Toxic effect of arsenite on the expression of liver multidrug resistance associated protein 2 in rat]. AB - OBJECTIVE: To investigate the role of multidrug resistance-associated protein 2 (MRP2) in the hepatic cell membrane of rats. METHODS: Thirty healthy Wistar rats were divided randomly into six groups based on time of administration (2 w, 4 w, 6 w) of 20 mg/kg of sodium arsenite, and their corresponding control groups. Animals were administered every other day. Arsenic content in blood and bile were detected by atomic absorption spectroscopy (AAS), and the expression of MRP2 in the membrane of hepatocyte by Western blotting was determined. RESULTS: Total arsenic levels (including organic arsenic and inorganic arsenic) in blood and bile were significantly higher than control groups (P < 0.05) at all three different time points, especially in 2 w and 4 w group (16.8 and 13.8 fold greater than that in control). The expression of MRP2 increased 36.61%, 32.36%, 12.73% more respectively in 2 w, 4 w, 6 w groups than those in control groups (P < 0.05). The expression of MRP2 was correlated with total arsenic content in bile (r = 0.713, P < 0.05). CONCLUSIONS: Bile is one of the major routes for the excretion of arsenite and its metabolites, and the overexpression of MRP2 may play an important role in the bile excretion of them at early stage. PMID- 15355705 TI - [Effect of low intensity and very high frequency electromagnetic radiation on occupationally exposed personnel]. AB - OBJECTIVE: To investigate the effect of low intensity and very high frequency (VHF) electromagnetic radiation (170 MHz) on nervous system function and serum enzymes and immune function in human subjects with occupational exposure to VHF. METHODS: To measure the intensity of VHF and other environmental factors on the spot, to hold the questionnaire about chief complaints, to examine the rheoencephalography and the neurobehavior function, to analyze ALT, AST, ALP and LDH, and IgA, IgM and IgG in experimental group and control group. RESULTS: The intensity of VHF (direction of antenna: 0 degrees, 10 m and 135 degrees, 20 m) was higher than that of national standard on-the-spot. The incidences of symptoms such as headache, insomnia and amnesia etc. was significantly higher in experimental group (P < 0.01). Rheoencephalography indicated that the raising time of both left [(0.155 3 +/- 0.057 9) s] and right [(0.154 1 +/- 0.059 2) s] in the experimental group after exposure were significantly longer than before exposure [(0.104 4 +/- 0.030 2) s, (0.103 2 +/- 0.030 4) s respectively] or in the control [(0.118 5 +/- 0.056 8) s, (0.117 7 +/- 0.057 5) s respectively, (P < 0.01)]. Neurobehavior function test showed that digital symbol, digital span and pursuit aiming test were decreased after exposure in the experimental group (P < 0.01). Serum enzyme analysis showed that AST, ALP and LDH were significantly increased after exposure in the experimental group (P < 0.01). No marked change was found in IgA level, while the levels of IgM and IgG after exposure in the experimental group especially the latter were significantly increased (P < 0.01). CONCLUSIONS: Low-intensity VHF radiation can decrease the nervous system function in occupationally exposed personnel and induce increase in some kinds of enzymes and immunoglobulins. PMID- 15355706 TI - [Study on the relationship between the level of urinary monomethylfomamide and the injury of liver and kidney in workers exposed to dimethylfomamide]. PMID- 15355707 TI - [The X-ray characteristics of rush pneumoconiosis]. PMID- 15355708 TI - [Investigation on the pneumosilicosis in small cast steel factories]. PMID- 15355709 TI - [Changes of serum protein in patients with acute organophosphorus pesticide poisoning]. PMID- 15355710 TI - [Changes and significance of urinary N-acetyl-beta-D-glucosaminidase in patients with acute organophosphorus pesticide poisoning]. PMID- 15355711 TI - [Investigation on the morbidity and mortality of pneumosilicosis in a quartz factory]. PMID- 15355712 TI - [Epidemiological investigation on the chronic diseases among professional cooks]. PMID- 15355713 TI - [The toxic effect of methamidophos and acephate on intracellular free Ca2+ and cAMP concentration in rat brain tissue]. PMID- 15355714 TI - [Effect of 2,4-D isooctylester on sperms in tail of epididymis and HSP70 expression of testis tissue in rats]. PMID- 15355715 TI - [Protective effect of vitamin C on muscle strain injury-induced peroxidative damage in rats]. PMID- 15355716 TI - [Evidence-based medicine and occupational health practice]. PMID- 15355718 TI - [Profile of the study of methods for detecting selenium]. PMID- 15355717 TI - [Application and advance of benchmark dose on risk assessment]. PMID- 15355719 TI - [Retrospect of occupational health and occupational disease statistical report]. PMID- 15355720 TI - [Application of care methods in monitoring of occupational injuries]. PMID- 15355721 TI - [A survey of the prevention of hazards of occupational disease in new projects founded during 1996 to 2001 in town-owned enterprises in Tianjin]. PMID- 15355722 TI - [Determination of lead in urine by hydride generation atomic fluorescence spectrometry]. PMID- 15355723 TI - [Analytical method for benzene, toluene, xylene, hexane, trichloroethylene in gooey agent by gas chromatography]. PMID- 15355724 TI - [Determination of total volatile organics in the air by gas chromatography]. PMID- 15355725 TI - [Determination of 23 drugs by capillary gas chromatography]. PMID- 15355726 TI - [Study on the health standard for phosphorus pentasulfide in the workshop air]. PMID- 15355727 TI - [Three cases of hypothyroidism induced by cosmetics containing mercury]. PMID- 15355728 TI - [Clinical analysis on 53 cases of occupational noise-induced hearing loss]. PMID- 15355729 TI - [Investigation on an accident of nitrogen mustard poisoning]. PMID- 15355730 TI - [Analysis of two accidents of working in pile wells]. PMID- 15355731 TI - [Analysis of three accidents of occupational latent H2As poisoning]. PMID- 15355732 TI - [Accuracy calculation for years of potential life lost caused by injuries]. PMID- 15355733 TI - [Study on the hygienic estimation of cold intensity in workers]. PMID- 15355734 TI - [How often does decreased consciousness of hypoglycaemia occur in children and adolescents with diabetes type 1 and what are its consequences?]. AB - The inability of the patient to recognize the risk of hypoglycemia is a very frequent phenomenon, but it is also often an underestimated complication in diabetes treated with insulin. The results of DCCT trial revealed that intensification in insulin therapy increases three-fold the risk of severe hypoglycaemia. Feeling the state of hypoglycaemia is the basic defensive mechanism in patients with diabetes type 1, making possible to start the self treatment. The decreased consciousness of hypoglycaemia makes limitations to intensive insulin therapy, which main aim is to stop later complications. THE AIM OF THE STUDY was to answer the questions: 1. How often does lack of consciousness of hypoglycaemia occur in children and adolescents with diabetes type 1. 2. What are the possible factors influencing appearance of hypoglycaemia. 3. Is lack of hypoglycaemia consciousness of a risk factor for severe hypoglycaemia. MATERIAL AND METHODS: The study was carried out on 318 patients aged x=13.6 yrs (4-21), suffering from diabetes, mean 6.6 yrs (2-18). The study was retrospective taking into consideration the period from 1.01.1998 to 31.12.2002. RESULTS: In the analysis of the questionnaire assessing the occurrence of hypoglycaemia it was found that 82 patients (25.8%) have problems with feeling the state of hypoglycaemia. We analyzed the influence of time of lasting diabetes and we found that patients with a longer duration of the disease more frequently have problems with feeling hypoglycaemia, 57% patients with lack of hypoglycaemia consciousness have bad metabolic control of the disease. In the analyzed period, 64 incidences of severe hypoglycaemia in 48 patients (30 boys and 18 girls) were found. In patients with lack of consciousness of feeling hypoglycaemia the incidences of severe hypoglycaemia occurred ten times more frequently compared to patients who feel hypoglycaemia. Sleeping makes it impossible to perceive early symptoms of hypoglycaemia: in our patients 51 severe incidences (79.7%) occurred at 1.00-3.00 a.m., 6 (9.3%) occurred at daybreak and 7 (11%) in the evening. CONCLUSIONS: 1. In patients with diabetes type 1 the lack of hypoglycaemia consciousness occurs in about 25%. 2. The lack of hypoglycaemia consciousness is closely connected with time of diabetes duration and with recurrence of hypoglycaemia incidences. 3. Patients with lack of hypoglycaemia consciousness are at greater risk for severe hypoglycemia. PMID- 15355735 TI - [Changes in concentrations of circulating insulin-like growth factors I and II and binding proteins BP-2 and BP-3 in children treated for acute lymphoblastic leukaemia]. AB - We determined serum levels of insulin-like growth factors (IGF-1, IGF-2) and binding proteins (IGFBP-2 and IGFBP-3) at diagnosis (I), during (II) and after therapy (III) in 28 children treated for ALL. RESULTS: 1) Serum absolute levels of IGF-1, IGF-2, IGFBP-3 rose and IGFBP-2 - decreased during analysis. 2) Mean values of IGF-1 SDS were similar at diagnosis, during and after treatment ( 1.78+/-0.9 vs. -1.86+/-1.36 vs. -1.8+/-0.83). IGF-2 SDS rose from 0.06+/-1.45 (I) to 0.64+/-1.4 (II) and to 0.83+/- 2.01 (III) and IGFBP-3 SDS increased from 0.23+/-2.23 (I) to 0.92+/-1.95 (II) and to 1.76+/-2.43 (III). IGFBP-2 SDS were elevated at diagnosis, during and after treatment 11.94+/-9.42 (I) and 10.58+/ 7.37 (II) and to 7.90+/-7.20 (III). 3) We observed positive correlations between: a) IGF-1 and IGFBP-3 at diagnosis (r=0.58 p=0.0001), during (r=0.69 p=0.0001) and after treatment (r=0.79 p=0.00001), b) IGF-1 and IGF-2 at diagnosis (r=0.57 p=0.00001) and after the end of therapy (r=0.48 p=0.01) c) IGF-2 and IGFBP-3 at diagnosis (r=0.7 p=0.0004) and after treatment (r=0.64 p=0.003) and d) negative correlation between IGF-1 and IGFBP-2 - at diagnosis (r=0.38 p=0.02) and after treatment (r=0.43 p=0.02). CONCLUSIONS: Decreased serum IGF-1 and elevated IGFBP 2 were observed at diagnosis and during treatment for ALL suggesting that especially IGFBP-2 may be related to the proliferation of lymphoblasts. PMID- 15355736 TI - [Ultrasonographic evaluation of atherosclerotic changes in carotid and brachial arteries in children with type 1 diabetes]. AB - INTRODUCTION: According to very well documented onset of atherosclerosis in early childhood, scientists are looking for good diagnostic methods for evaluating first changes in arterial blood vessels noninvasively. We want to know more about the pathogenetic mechanisms and about changes in vessels especially in group of young people with risk factors of premature atherosclerosis. The significance of endothelial dysfunction in very early phase of this process is known very well so far. High resolution echocardiography seems to be a good method which allows to examine arteries in children and adolescents. Because of localization, brachial and carotid arteries are very good field for this kind of examinations. THE AIM of this study was the evaluation with high resolution echocardiography of endothelial function in type 1 diabetes children and adolescents. We also measured the intimal plus medial thickness in carotid communis arteries (IMT). MATERIAL AND METHODS: We examined a group of 64 children (29 boys and 35 girls) aged 14.8-16.2 yr (mean 15.5 yr) suffering from diabetes type 1. The control group consisted of 24 children (9 boys, 15 girls) in similar age. Using high resolution echocardiography, B-mode images, we measured in the diastole phase, distance "m-m" in brachial arteries (distance between two "m" lines which are borders among media and adventitia of near and far wall of the artery) at rest, during reactive hyperaemia (with increased flow causing endothelium-dependent dilatation FMD), again at rest and after sublingual glyceryl trinitrate (causing endothelium-independent dilatation NTGMD). Using Doppler technic we evaluated baseline flow and calculated the degree of reactive hyperemia. We also measured intimal plus medial thickness in every carotid artery three times and calculated mean value. In our analysis we estimated the concentrations of cholesterol, HDL- cholesterol, LDL-cholesterol and triglycerides. RESULTS: We noticed higher IMT values in the whole examined group compared to the control group (0.52 mm vs. 0.43 mm, p<0.05). In diabetic children the vessel size was similar to control group but FMD was significantly impaired (5.56% vs. 9.47%, p<0.05). The level of triglycerides and total cholesterol was higher in the examined group. CONCLUSIONS: 1. The evaluation of IMT in the carotid arteries in patients with type 1 diabetes showed a more advanced degree of atherosclerotic changes in this group compared to healthy controls. 2. FMD evaluated in brachial artery is a usefull tool in assessing impaired endothelial function in people suffering from the risks factors of atherosclerosis. 3. Ultrasonographic methods of evaluating atherosclerotic changes in arterial vessels should be more often used in practice as relatively easy, noninvasive and inexpensive. PMID- 15355737 TI - [Prevalence of chronic diabetes complications depending on the method of insulin therapy in children and adolescents with type 1 diabetes]. AB - The critical study showing advantages of intensive insulin therapy in comparison with the classic method was Diabetes Control and Complications Trial. Conclusion of that study was unequivocal: hyperglycemia has a significant influence on the development of the chronic diabetes complications. THE AIM of this study was to estimate the influence of the conventional insulin therapy in the past and its duration on the development of complications in patients suffering from type 1 diabetes for more than 5 years. MATERIAL AND METHODS: 185 patients aged 16.7 years (+/-3.4) with diabetes onset in the age 8.4 years (+/-3.8) participated in this study. There were 84 girls and 101 boys. More of them were treated by classic insulin therapy at the beginning of disease (n=148). 37 persons had intensive insulin therapy all the time. RESULTS: The presence of at least one complication was found in 49 persons (26.5%), two complications - in 14 patients (7.57%) and three complications in 4 patients (2.16%). The conventional therapy duration was twice longer in patients with retinopathy. The significant influence of the age, the age of diabetes onset and duration was found in those patients. Comparing the groups of patients treated conventionally less than 3.5 years and more than 3.5 years it was found that patients in the first group were older, fell ill later and they had longer diabetes duration than patients in the second group. The risk of retinopathy increases from 4.2% to 8.9% in patients treated conventionally more than 3.5 years. The risk of diabetic cataract is 5 times higher in that group. CONCLUSIONS: Chronic complications of diabetes occur in 26.5 % patients with a diabetes duration of at least 5 years. CIT is the risk factor of the eye complications in diabetes, particularly when applied longer than 3.5 years. The prolongation of CIT over 3.5 years increases the risk of retinopathy twice and the risk of cataract 5 times in patients with type 1 diabetes. So the early intensification of insulin therapy reduces the risk of the retinopathy and cataract development in later course of the disease. PMID- 15355738 TI - [Assessment of growth factor levels in adolescents with type 1 diabetes mellitus and the beginning of diabetic microangiopathy]. AB - OBJECTIVE: The purpose of the study was the evaluation of growth hormone (GH), insulin-like growth factor (IGF-1), insulin-like growth factor binding protein (IGFBP-3) and vascular endothelial growth factor (VEGF) in adolescents with type 1 diabetes as well as the relationship between the concentration of examined proteins and the early development of diabetic microangiopathy. RESEARCH DESIGN AND METHODS: The study included 68 patients aged 15.54+/-2.9 years with type 1 diabetes. They were divided into groups: I - adolescents with the beginning of vascular complications, II - without complications. Controls consisted of healthy adolescents. GH, IGF-1, IGFBP-3 and VEGF were determined with use of ready kits. RESULTS: The increased levels of GH were found in diabetic adolescents in comparison with controls (10.11+/-16.21 vs. 2.89+/-4.03 micro IU/ml, p<0.05). The levels of IGF 1 were lower in adolescents with type 1 diabetes than in controls (283.48+/-117.36 vs. 427.95+/-177.48 ng/ml, p<0.05), as well as the levels of IGFBP-3 (5555.21+/-1158 vs. 6622.18+/-1110 pg/ml, p<0.05). Significantly higher concentration of VEGF was revealed in diabetic adolescents in comparison with controls (328.68+/-251 vs. 132.19+/-85 pg/ml, p<0.05). The highest levels of VEGF were reported in diabetic patients with retinopathy in comparison with patients without complications. CONCLUSIONS: The impaired activity of GH-IGF-1-IGFBP-3 axis may be responsible for the development of diabetic microangiopathy. The evaluation of VEGF concentration can be a sensitive early marker of microangiopathy. PMID- 15355739 TI - [Sudden death caused by myocarditis in a 14-year old boy with type I diabetes]. AB - We report a case of diabetic ketoacidosis complicated by acute myocarditis which was confirmed by necroscopy. A 14 year boy was hospitalised with severe ketoacidosis. The patient levels of creatinine, transaminases were elevated, symptoms of hearth disease were complicated by dehydration and high temperature. An episode of upper respiratory viral infection before the onset of acute diabetes suggested that the patient died from viral myocarditis. PMID- 15355740 TI - [Leptin, body composition and bone mineralization in children after treatment for Wilms tumor]. AB - Advances in diagnosis and improved methods of treatment have resulted in increasing number of long-term survivors in children with Wilms tumor. Growth and puberty are important for accumulation of bone mass; chemotherapy nad radiotherapy used in treatment for Wilms tumor can influence bone structure and physical development. Leptin plays an important role in metabolism of adipose tissue and bone mineralization. Considering that neoplasm and its treatment can affect normal development in childhood, we analysed the influence of antineoplastic treatment on bone mineralization and the correlations between serum leptin levels, body composition and bone mineral density in survivors of Wilms tumor. Twenty subjects (12 boys) treated for Wilms tumor at the mean age of 10.9 (range 3-20 years) participated in this study. Mean follow up period after discontinuation of therapy was 5.6 years (range 2 months - 13.5 years). Mean age of diagnosis was 3.9 years (range 1 month - 12.6 years). 18 patients received chemotherapy, 7 - additionally radiotherapy and 2 infants had only surgery treatment. We measured fat mass - FM, fat free mass - FFM, bone mineral density - BMD total and BMD spine using dual energy x-ray absorptiometry (DXA) and compared to the results obtained for healthy references (SD score). Leptin levels were measured with RIA method. RESULTS: 1. No difference was found in leptin levels, body mass index, FM, FFM, BMD total and spine in relation to sex. 2. Means of SDS BMI, FM, FFM, BMD and leptin were in the normal range for the age and sex matched controls. 3. We found the correlation between leptin level and BMI, FM, FFM and BMD total and spine, no correlation was found between SDS values. 4. We observed a positive correlation between SDS BMD and SDS BMI, FM, FFM, BMD spine. 5. BMI, FM and leptin levels were higher in children treated with radiotherapy and chemotherapy than in children treated with only chemotherapy. However, the SDS values were comparable with the healthy controls. 6. SDS BMD total was decreased in 5/20 subjects (25% of all studied patients) compared with healthy controls. CONCLUSIONS: The results demonstrated the risk of osteopenia in the group of children treated for Wilms tumor and the necessity for long-term monitoring of bone mineralization. PMID- 15355741 TI - Gene targeting: roadmap to future therapies. PMID- 15355742 TI - Etanercept and methotrexate combination in rheumatoid arthritis. PMID- 15355743 TI - Adalimumab in rheumatoid arthritis. PMID- 15355744 TI - Infliximab in rheumatoid arthritis. PMID- 15355745 TI - Targeting cytokines beyond tumor necrosis factor-alpha and interleukin-1 in rheumatoid arthritis. AB - Targeting tumor necrosis factor-a has proven of considerable value in treatment for rheumatoid arthritis, with substantial benefits achieved in a proportion of treated patients. However, a significant number of patients do not achieve sufficient improvement and as a result there remains considerable unmet clinical need. A number of cytokines have recently been described with proinflammatory activity in rheumatoid arthritis synovitis, including interleukin (IL) -6, IL-12, IL-15, and IL-18. We review recent data that support the notion that some or all of these moieties offer therapeutic potential. The possibility that some may be useful in partial responders to tumor necrosis factor blocking agents or in synergy with the latter is discussed. PMID- 15355746 TI - Update on autoantibodies in rheumatoid arthritis. AB - Detection of rheumatoid factor (RF) in the serum of patients with rheumatoid arthritis (RA) was one of the first indications of autoimmunity in RA. The role of RF in the diagnosis of RA has been well-documented, but it has suboptimal sensitivity and specificity. Although patients with RF-positive RA generally have more severe disease than those with RF-negative RA, RF is not a reliable predictor of disease severity in individual patients. Multiple other autoantibodies have been found in RA, with recent interest focused on those directed at cyclic citrullinated peptides. Panels of autoantibodies may ultimately prove useful in preclinical diagnosis and prediction of clinical course in patients with RA and other forms of arthritis, and provide insights into the pathogenesis of the disease. PMID- 15355747 TI - Aspirin, NSAIDs, and COX-2 inhibitors in cardiovascular disease: possible interactions and implications for treatment of rheumatoid arthritis. AB - Aspirin, nonsteroidal antiinflammatory drugs (NSAIDs), and cyclooxygenase-2 (COX 2) inhibitors are widely used in patients with rheumatoid arthritis. Aspirin has the largest and most persuasive body of randomized trial evidence to support its use in secondary prevention for cardiovascular disease (CVD) and primary prevention for myocardial infarction. There is, however, a possible deleterious interaction between aspirin and NSAIDs on CVD that requires further research. Aspirin, NSAIDs, and to a lesser extent COX-2 inhibitors are associated with increased gastrointestinal side effects and bleeding, alone and in combination. The more widespread and appropriate use of aspirin in patients with rheumatoid arthritis will avoid many premature deaths in secondary prevention for CVD and first myocardial infarctions in primary prevention. PMID- 15355748 TI - B cells: new ways to inhibit their function in rheumatoid arthritis. AB - The apparent efficacy of B-cell depletion in autoimmune diseases has increased interest in targeting B cells. One goal of next generation therapies is to develop treatments that block B-cell activation and preserve resting nonautoimmune cells that maintain B cell memory. To do so, one needs to understand how B cells are activated and what receptors and intracellular signaling pathways regulate this process. This paper will summarize B-cell activation pathways and illustrate how these are being targeted in the development of new treatments for rheumatoid arthritis. PMID- 15355749 TI - Occupational and environmental exposures as risk factors for systemic lupus erythematosus. AB - Although genetic susceptibility plays a strong role in the etiology of systemic lupus erythematosus (SLE), recent research has provided new evidence of the potential influence of environmental factors in the risk for this disease. This paper describes epidemiologic and experimental research pertaining to occupational and environmental sources of exposure to respirable crystalline silica, solvents and pesticides, and two "lifestyle" factors (smoking and hair dye use). As has been seen with other systemic autoimmune diseases (eg, systemic sclerosis and rheumatoid arthritis), a series of epidemiologic studies, using different designs in different settings, have demonstrated relatively strong and consistent associations between occupational silica exposure and SLE. The type and quality of exposure assessment is an important consideration in evaluating these studies. Recent experimental studies examined the effect of trichloroethylene exposure in MRL+/+ mice, but to date there have been few epidemiologic studies of solvents and SLE. There are numerous avenues with respect to environmental factors in SLE that need additional research. PMID- 15355750 TI - New advances in measurement of complement activation: lessons of systemic lupus erythematosus. AB - Activation of the complement system plays a fundamental role in the pathogenesis of systemic lupus erythematosus (SLE). For the past several decades, quantifying this process has focused primarily on determination of serum C3 and C4, although the utility of these assays for diagnosis and monitoring disease activity is still debated. During this same timespan, knowledge of the complement system has exploded, with identification of more than 30 proteins, an abundance of newly recognized functions, and even a third pathway of activation. These advances suggest that it is appropriate to revisit the complement system as a potential source of biomarkers for SLE. This paper reviews briefly the role of complement in SLE and other inflammatory diseases, discusses conventional methods for complement measurement and their drawbacks, and focuses on recent advancements in harnessing the complement system for monitoring SLE. Specifically, novel assays that measure cell-bound complement activation products are introduced and their utility as biomarkers of SLE disease activity is discussed. PMID- 15355751 TI - Biomarkers in systemic lupus erythematosus. AB - Despite the longstanding interest and large number of publications on biomarkers in lupus, there are no validated and widely accepted biomarkers of systemic lupus erythematosus to date. To achieve the ultimate goal, to have a biomarker as a surrogate endpoint in clinical studies, candidate biomarkers have to first be validated in a statistically rigorous way. However, to qualify as a surrogate endpoint, even validated biomarkers have to go through a process that demonstrates that they accurately reflect a clinically important outcome. These goals can only be achieved in large multicenter, properly conducted studies. We reviewed the difficulties involved in developing validated biomarkers for systemic lupus erythematosus and summarized the available data on the most promising biomarker candidates of disease susceptibility and disease activity. We also report on the current status of a multicenter initiative to concentrate efforts of biomarker development. PMID- 15355753 TI - Anticraving agents for the treatment of addictive disorders. PMID- 15355752 TI - Current advances in the human lupus genetics. AB - Genetic predisposition has been firmly established as a key element in susceptibility to systemic lupus erythematosus (SLE). During the past three decades, association studies have assessed many genes for potential roles in predisposing to SLE. These studies have identified a few risk factors including hereditary deficiency of complement components, major histocompatibility complex class II alleles, and allelic variants for the Fc portion of IgG (FCGR) genes. In recent years, a few groups have completed linkage analyses in data sets from families containing multiple members affected with SLE. Results from these initial genome scans are encouraging; approximately eight chromosomal regions have been identified exhibiting evidence for significant linkage to SLE and have been confirmed using independent cohorts (1q23, 1q25-31, 1q41-42, 2q35-37, 4p16 15.2, 6p11-21, 12q24, and 16q12), suggesting the high likelihood of the presence of one or multiple SLE susceptibility genes at each locus. Another approach of linkage analyses conditioned on pedigrees where one affected member manifesting a particular clinical condition has also identified many chromosomal regions linked to SLE. Within several established susceptibility loci, evidence for association of positional candidate genes is emerging. Within 2q35-37, an intronic single nucleotide polymorphism (SNP) of the positional candidate gene program cell death 1 gene has been associated with SLE susceptibility. The SLE-associated SNP affects a transcription factor, RUNX1, binding site. Recently, SNPs of novel positional candidate genes that influence RUNX1 binding motifs have also been associated with other autoimmune diseases, suggesting the possibility of a common theme shared among susceptibility genes for autoimmune diseases. In the coming years, susceptibility genes responsible for the observed linkage will be identified, and will lead to further delineating genetic pathways involved in susceptibility to SLE. PMID- 15355754 TI - Recent advances in the pharmacotherapy of cocaine dependence. AB - The pharmacotherapy of cocaine dependence is a rapidly developing field of research that may soon produce efficacious medications. Expanding research on reward-related brain circuitry, which is acutely activated and chronically dysregulated by cocaine, has helped reveal the neurobiological features of cocaine dependence and is guiding pharmacologic strategies that have significant potential to improve clinical outcome. Cocaine dependence is a multifaceted disorder with distinct clinical components that may respond to different pharmacologic approaches. Pharmacologic strategies for this disorder include blocking euphoria, reducing withdrawal and negative mood symptoms, ameliorating craving, and enhancing the prefrontal cortical function that seems to be impaired in cocaine-dependent patients. One medication may not be sufficient to treat these diverse elements of cocaine dependence because preliminary studies report efficacy with medications that have opposite actions on reward-related circuits. This review highlights pertinent advances in cocaine neurobiology, recent clinical trials, and controversies in the pharmacologic treatment of cocaine dependence. PMID- 15355756 TI - Recent advances in the treatment of opiate addiction. AB - Although addiction to heroin and other opiates is a major public health issue in the United States and many other countries, advances in pharmacologic and nonpharmacologic treatment have been made. Some of these advances represent a major shift from traditional treatment philosophies, whereas others are characterized by more subtle, though important, improvements. This review discusses recent advances in opiate-addiction treatment in the context of three main domains: the use of buprenorphine and buprenorphine/naloxone in an office based paradigm, psychosocial treatment and the addressing of medical and psychiatric comorbidity, and harm reduction strategies. PMID- 15355755 TI - Recent advances in the pharmacotherapy of alcoholism. AB - Alcoholism is a devastating illness that leads to great societal losses. Despite significant health consequences, there are few medically based treatments for alcoholism. During the past decade, a better understanding of the neuroscientific underpinnings of addiction has led to the use of novel pharmacotherapeutic treatments for alcoholism. In particular, there have been new developments in the understanding of the involvement of the dopamine, opiate, serotonin, gamma aminobutyric acid, and glutamate neurotransmitter systems in the pathophysiology of alcohol withdrawal, alcohol dependence, and in subtypes of individuals with alcoholism. In this article, new developments in the pharmacotherapy of alcohol dependence will be reviewed. In particular, the use of anticonvulsants in alcohol withdrawal and protracted abstinence syndromes will be discussed. Data on naltrexone, acamprosate, and topiramate will be highlighted. In addition, data concerning the use of serotonin reuptake inhibitors in subtypes of alcoholism and the use of combination pharmacotherapy will be reviewed. PMID- 15355757 TI - Recent understanding in the mechanisms of addiction. AB - The rewarding effects of drugs of abuse have been linked to increases in dopamine transmission. However, changes in brain chemistry and morphology that are produced in addiction underlie the long-lasting vulnerability to relapse and are more closely linked with the adaptations in excitatory transmission. The drug induced changes in excitatory transmission seem to be pathologic exacerbations of normal forms of brain plasticity, and they occur in the brain areas linked by neuroimaging studies in addicted patients to craving and relapse. This review describes the brain adaptations produced in excitatory transmission by addictive drugs and identifies new potential sites of pharmacotherapeutic intervention to ameliorate addiction. PMID- 15355758 TI - Neurosurgical interventions for neuropsychiatric syndromes. AB - Psychosurgical procedures have been used for the treatment of intractable mental illness for more than 50 years. With improvements in surgical techniques, including new implantable stimulators, advances in functional neuroimaging, and progress in our fundamental understanding of the pathophysiology of mental illness there is a renewed interest in neurosurgical treatment of refractory psychiatric illness. This article will review the history of psychosurgery and recent developments in surgical techniques and implantable devices used in this context. The results of psychosurgery for the treatment of several psychiatric conditions and neuropsychiatric symptoms will be presented, including obsessive compulsive disorder, Tourette's syndrome, depression, anxiety, aggression, self injurious behavior, and schizophrenia. Lastly, a perspective on the current and future role of psychosurgery for the treatment of mental illnesses will be discussed. PMID- 15355759 TI - Cognitive effects of cytotoxic cancer chemotherapy: predisposing risk factors and potential treatments. AB - Increasing evidence suggests that systemic cancer chemotherapy can have significant long-term effects on cognition, particularly on verbal learning, memory, attention, and speed of information processing. These deficits can be a source of significant distress to survivors. There is much less known about the mechanisms, predisposing vulnerabilities, and treatment of these deficits. We will summarize current knowledge of chemotherapy-associated cognitive deficits. Emerging theories about the role of selected genetic polymorphisms in heightening the vulnerability to chemotherapy-induced cognitive decline will be described. PMID- 15355760 TI - Viral encephalitis: neuropsychiatric and neurobehavioral aspects. AB - Viral encephalitis, a condition in which a virus infects the brain and produces an inflammatory response, affects approximately 20,000 individuals per year in the United States. The viral encephalidities include sporadic and epidemic acute viral encephalidities and subacute and chronic/progressive viral encephalitis or encephalomyelitis. In people who survive these conditions, postencephalitic impairments of elemental neurologic, cognitive, emotional, and behavioral function are common. This article will provide a brief overview of the diagnosis and acute management of acute viral infections of the central nervous system. The neurologic and neuropsychiatric features, neuropathologies, and treatments of two of the more common types of acute viral encephalitis in North America--herpes simplex encephalitis and West Nile encephalitis--will be reviewed. The current and future role of psychiatrists and neuropsychiatrists in the care and study of individuals with these conditions will be discussed. PMID- 15355761 TI - Neuropsychiatric syndromes in adults with intellectual disability: issues in assessment and treatment. AB - The purpose of this article is to present a survey of important neuropsychiatric issues and recent findings regarding the evaluation and treatment of neuropsychiatric symptoms and syndromes in patients with intellectual disability (ID). The cause of ID, environmental or genetic, can be determined in few patients. Etiology is idiopathic in most patients. ID and psychiatric illness are not mutually exclusive; individuals with ID have increased rates of psychiatric illness. Although recognition of significant axis I psychopathology is important, not all challenging behaviors in persons with ID will have a clear axis I diagnosis. Psychologic, behavioral, and environmental treatments are appropriate measures, but pharmacotherapy often is needed. Our experience has shown us that the more severe, disruptive, and dangerous the behavior, the stronger the indication for empirical drug treatment trials. Community-based models of support with neuropsychiatric intervention can be a potent therapeutic combination in the management of challenging behaviors in individuals with ID. PMID- 15355763 TI - A model for the function of sperm DNA degradation. AB - In this review, we present our recent evidence suggesting, but not yet proving, that mammalian spermatozoa contain a mechanism by which they can digest their own DNA when exposed to a stressful environment. We discuss our recent data that demonstrate that when mammalian spermatozoa are treated in a variety of ways, the paternal chromosomes in the zygote, or the sperm DNA itself, are degraded into large, chromosome-sized fragments. These published data support the existence of nuclease activity in spermatozoa. We suggest that this nuclease activity is part of a mechanism the spermatozoon uses when it encounters a stressful environment to prevent fertilisation and to avoid the transmission of potentially damaged DNA to the embryo. We propose a model based on sperm chromatin structure by which this nuclease can digest the highly condensed sperm chromatin. PMID- 15355762 TI - The neural circuitry of reward and its relevance to psychiatric disorders. AB - Scientific interest in how the brain processes reward has burgeoned during the past 50 years since the discovery that rats will do tasks such as pressing a lever to obtain electrical stimulation of the brain. This interest was additionally encouraged by the observation of an association between reward and dopamine activity in the mesocorticolimbic system. In this article, we will discuss the complex nature of reward processing and recent animal studies and human functional neuroimaging studies to elucidate the current understanding of the neural substrates of reward processing and its components. Lastly, we will review recent theoretical and empirical work investigating the role of brain reward circuitry in several psychiatric disorders, including substance use disorders, schizophrenia, pathologic gambling, major depressive disorder, and attention-deficit/hyperactivity disorder. PMID- 15355765 TI - [Interventional strategies for the rational use of antimicrobial drugs outside the hospital setting]. PMID- 15355766 TI - [Etiology of acute otitis media in a children's hospital and antibiotic sensitivity of the bacteria involved]. AB - BACKGROUND: This study provides an update on the bacterial etiology of acute otitis media in our area, the antibiotic sensitivity of the bacteria implicated in this condition, and the prevalence of circulating Streptococcus pneumoniae serotypes. METHODS: Results from a total of 240 samples obtained by diagnostic tympanocentesis and 167 samples of spontaneous otorrhea processed between 1999 and 2001 were reviewed retrospectively. RESULTS: The mean age of the patients studied was 17 months and the median was 13 months (range: 1 month-7 years). Among the tympanocentesis samples, S. pneumoniae was recovered from 67 (27.9%), Haemophilus influenzae from 60 (25%), both S. pneumoniae and H. influenzae from 3 (1.3%) and Moraxella catarrhalis from 6 (2.5%). Among the spontaneous otorrhea samples, S. pneumoniae was recovered from 15 (9%), H. influenzae from 25 (15%) and both S. pneumoniae and H. influenzae from 1 (0.6%). The remaining samples showed either no growth or recovery of colonizing flora. The main findings were as follows: 49.3% of S. pneumoniae strains showed intermediate sensitivity to penicillin (MIC: 0.12-1 microg/ml), 16.9% were resistant to penicillin (MIC: > or = 2 microg/ml) and 54% were resistant to macrolides; 24,7% of H. influenzae and 100% of M. catarrhalis strains were beta-lactamase producers; and 64 (84.2%) of 76 S. pneumoniae serotyped strains belonged to pneumococcal heptavalent vaccine serotypes. CONCLUSIONS: S. pneumoniae and H. influenzae were the main causal agents of acute otitis media. Antibiotic sensitivity of the bacteria involved showed the same characteristics as the general pattern in our country. Spontaneous otorrhea culture was not a useful method for establishing the etiology of acute otitis media. Knowledge of the distribution of S. pneumoniae serotypes is essential for assessing epidemiological changes resulting from the use of heptavalent pneumococcal vaccine. PMID- 15355767 TI - [Evaluation of xylose galactosidase (XG) culture medium for the isolation of enteropathogens]. AB - INTRODUCTION: The chromogenic medium, XG, was evaluated and compared to conventional media for the isolation of Salmonella spp., Shigella spp., Yersinia enterocolitica and Aeromonas spp. METHODS: A total of 1226 human stool samples were inoculated on XG, MacConkey agar, Salmonella-Shigella agar (SS), selenite broth, blood-ampicillin agar and cefsulodin-Irgasan-novobiocin agar (CIN). RESULTS: The 235 positive cultures included the following: 229 Salmonella spp., 3 Shigella spp., 2 Yersinia enterocolitica and one Aeromonas spp. Among the 229 containing Salmonella spp., 100 were detected on both XG and conventional media and the 129 remaining were detected only on conventional media; recovery of Salmonella spp. on conventional media was significantly higher with respect to XG medium (p < 0.005). The 3 isolates of Shigella spp. were obtained on XG, the 2 isolates of Yersinia enterocolitica were recovered on CIN agar and the single isolate of Aeromonas spp. was obtained both on XG and blood-ampicillin agar. Colonies suspected to be some of the enteropathogens investigated were present in 791 of the negative stool samples. Among these false-positives 441 (35.9%) were obtained from XG, 142 (11.6%) after selenite enrichment, 132 (10.8%) from MacConkey agar and 76 (6.2%) from SS agar. Most of the false-positive isolates obtained on XG medium were consistent with Salmonella spp. (n = 408). CONCLUSIONS: XG chromogenic medium showed low sensitivity (64%) and specificity (69%) for the detection of Salmonella spp. Recovery of Shigella spp. on XG medium in three samples may have been due to the immediate processing of the samples. We conclude that XG chromogenic medium can not be recommended as an alternative to currently used conventional media. PMID- 15355768 TI - [Analysis of antibiotic use in the Community of Valencia (2000-2002)]. AB - OBJECTIVE: To quantify antibiotic prescriptions in the Autonomous Community of Valencia in the years 2000-2002 and analyze the differences in antibiotic use among 20 Valencian Community health areas. METHODS: Retrospective observational population study (3,983,464 inhabitants) of prescriptions recorded in Valencian Community pharmacies during the period 2000-2002. We calculated defined daily doses per 1000 inhabitants and day (DHD), for the population without pharmaceutical fees (e.g. pensioners), the population with pharmaceutical fees (e.g. actively working) and the total population. The antibiotics analyzed belonged the J01 group (ATC/DDD classification). RESULTS: Antibiotic consumption was 26.47 DHD in 2000, 23.56 DHD in 2001 and 23.48 DHD in 2002. The most frequently prescribed drug groups included: broad-spectrum penicillins (J01C) 11.8 DHD, macrolides (J01F) 5.6 DHD, and other beta-lactams (J01D) 3.95 DHD. There was considerable variation between the population without pharmaceutical fees and the population with pharmaceutical fees: JO1C (DHD 18.49 vs. 9.73), JO1F (11.06 vs. 3.84) and quinolones (J01M) (9.16 vs. 1.22) (p < 0.05). There was also substantial variation between areas, both overall and by groups (p < 0.05): J01C 6.36 DHD to 12.14 DHD; (J01F), 3.78 DHD to 6.08 DHD; (J01D), 2.28 DHD to 4.65 DHD, and (J01M) 2.39 DHD to 3.6 DHD. CONCLUSIONS: There was a slight trend to decreasing antibiotic use in the Community of Valencia over the period of 2000 2002. The population without pharmaceutical fees showed much higher antibiotic use than those with pharmaceutical fees, and there was considerable variation in prescriptions between the different public health care areas. PMID- 15355769 TI - Assessment of three rapid methods for the detection of methicillin-resistant Staphylococcus aureus. AB - We evaluated three rapid methods to detect methicillin-resistant Staphylococcus aureus (MRSA) and compared them with PCR amplification of mecA. A total of 103 S. aureus strains were studied by MRSA-Screen, BBL Crystal, Velogene Genomic and mecA PCR. All the methods detected the 61 MRSA strains having the mecA gene, showing 100% sensitivity and specificity. Despite the correlation between all the rapid methods and PCR, the ease of use and shorter turnaround time of MRSA-Screen were important factors leading to the selection of this method as the routine screening technique for MRSA. PMID- 15355770 TI - [Up to date in sexually transmitted infections: epidemiology, diagnostic approaches and treatments]. AB - In the last years, there have been important advances in sexually transmitted infections such as genome sequencing of Treponema pallidum, Chlamydia trachomatis or Mycoplasma genitalium; the new taxonomic position of Calymmatobacterium granulomatis; commercial diagnostic systems based on nucleic acid amplification; the emergence of quinolone resistance in Neisseria gonorrhoeae; new therapeutic approaches in vulvovaginal candidiasis that include boric acid; the demonstration that valacyclovir reduces the risk of transmission of genital herpes or the availability of immune-response modifier in the treatment of genital warts, and that are questions in the goal of this review. Viral hepatitis and HIV were no reviewed by space reasons. PMID- 15355771 TI - [Avian influenza A (H5N1) infectious in both birds and humans in South-Eastern Asian countries]. AB - Avian influenza affects most types of birds and occurs in epidemics on poultry farms. The fatal disease is named "highly pathogenic avian influenza" and is caused by influenza A virus subtypes H5 and H7. The natural reservoir is the migratory waterfowl that occasionally infects domestic poultry. In 1997 in Hong Kong, 18 persons were infected and 6 of them died. At the end of 2003 and the beginning of 2004, avian influenza H5N1 infected numerous farms in several South Eastern Asian countries. The virus was transmitted to humans in close contact with infected birds. A total of 34 persons were infected and 23 of them died. There is currently a considerable concern about the H5N1 avian influenza that has infected humans: the high virulence, evolution rate, the possibility of recombination with other influenza viruses, how H5N1 variants that infect humans or different approaches to the development of influenza vaccines. PMID- 15355772 TI - [Epidemiology of HIV/AIDS infection and relationship with other sexually transmitted diseases (STD). Future prospects]. AB - Over the last decade, different cross-sectional and case-control studies have shown the relationship between HIV and Sexually Transmitted infections (STI). In this sense, different reasons are found: a modification of the clinical manifestations of some STI as well as an increase of HIV infectiousness and susceptibility to HIV. The main objective of HIV/AIDS and STI surveillance includes: to assess its temporal trends, to define high risk groups and high-risk behaviours and to plan and to evaluate public health programmes. Nevertheless, classical surveillance systems are not enough to describe the real situation of HIV/AIDS and STI. Thus, an enhanced surveillance system which includes complementary resources should be implemented. PMID- 15355773 TI - [Recurrent angioedema in a young woman]. PMID- 15355774 TI - [Post-trauma ocular pain and delayed blindness in a patient living in a rural setting]. PMID- 15355775 TI - [Home hospitalization and community-acquired pneumonia]. PMID- 15355778 TI - [Analysis of hospital admittance and mortality in AIDS patients]. PMID- 15355781 TI - [Endometrial tuberculosis in postmenopausal patients]. PMID- 15355782 TI - [Prevalence of genital infection due to Chlamydia trachomatis in the general population and in a group of prostitutes in Palma de Mallorca]. PMID- 15355783 TI - [Tuberculous ulcer in the hard palate of a patient with human immunodeficiency virus (HIV) infection]. PMID- 15355784 TI - Eggs over easy: cell death in the Drosophila ovary. AB - Programmed cell death is the most common fate of female germ cells in Drosophila and many animals. In Drosophila, oocytes form in individual egg chambers that are supported by germline nurse cells and surrounded by somatic follicle cells. As oogenesis proceeds, 15 nurse cells die for every oocyte that is produced. In addition to this developmentally regulated cell death, groups of germ cells or entire egg chambers may be induced to undergo apoptosis in response to starvation or other insults. Recent findings suggest that these different types of cell death involve distinct genetic pathways. This review focuses on progress towards elucidating the molecular mechanisms acting during programmed cell death in Drosophila oogenesis. PMID- 15355785 TI - Telomere biology in mammalian germ cells and during development. AB - The development of an organism is a strictly regulated program in which controlled gene expression guarantees the establishment of a specific phenotype. The chromosome termini or so-called telomeres preserve the integrity of the genome within developing cells. In the germline, during early development, and in highly proliferative organs, human telomeres are balanced between shortening processes with each cell division and elongation by telomerase, but once terminally differentiated or mature the equilibrium is shifted to gradual shortening by repression of the telomerase enzyme. Telomere length is to a large extent genetically determined and the neonatal telomere length equilibrium is, in fact, a matter of evolution. Gradual telomere shortening in normal human somatic cells during consecutive rounds of replication eventually leads to critically short telomeres that induce replicative senescence in vitro and probably in vivo. Hence, a molecular clock is set during development, which determines the replicative potential of cells during extrauterine life. Telomeres might be directly or indirectly implicated in longevity determination in vivo, and information on telomere length setting in utero and beyond should help elucidate presumed causal connections between early growth and aging disorders later in life. Only limited information exists concerning the mechanisms underlying overall telomere length regulation in the germline and during early development, especially in humans. The intent of this review is to focus on recent advances in our understanding of telomere biology in germline cells as well as during development (pre- and postimplantation periods) in an attempt to summarize our knowledge about telomere length determination and its importance for normal development in utero and the occurrence of the aging and abnormal phenotype later on. PMID- 15355786 TI - Regulatory function of whey acidic protein in the proliferation of mouse mammary epithelial cells in vivo and in vitro. AB - Although possible biological functions of whey acidic protein (WAP) have been suggested, few studies have focused on investigating the function of WAP. This paper describes evidence for WAP function in lobulo-alveolar development in mammary glands in vivo and in the cell cycle progression of mammary epithelial cells in vitro. Ubiquitous overexpression of WAP transgene impaired only lobulo alveolar development in the mammary glands of transgenic female mice but not other physiological functions, indicating that the inhibitory function of WAP is specific to mammary alveolar cells. The forced expression of WAP significantly inhibited the proliferation of mouse mammary epithelial cells (HC11 cells and EpH4/K6 cells), whereas it did not affect that of NIH3T3 cells. Co-culturing of WAP-clonal cells and control cells using a transwell insert demonstrated that WAP inhibited the proliferation of HC11 cells through a paracrine action but not that of NIH3T3 cells, and that WAP was able to bind to HC11 cells but not to NIH3T3 cells. Apoptosis was not enhanced in the HC11 cells with stable WAP expression (WAP-clonal HC11 cells). BrdU incorporation and FACScan analyses revealed that cell cycle progression from the G0/G1 to the S phase was inhibited in the WAP clonal HC11 cells. Among G1 cyclins, the expression of cyclin D1 and D3 was significantly decreased in the WAP-clonal HC11 cells. The present results provide the first documented evidence that WAP plays a negative regulatory role in the cell cycle progression of mammary epithelial cells through an autocrine or paracrine mechanism in vivo. PMID- 15355787 TI - PKC isotypes in post-activated and fertilized mouse eggs: association with the meiotic spindle. AB - Several isotypes of protein kinase C (PKC) have been reported to be expressed in mammalian eggs, but it is unknown whether these isotypes have a common function in the egg during or within the first few hours of fertilization. Here we show that the isotypes of PKC exhibit distinct patterns of enrichment immediately after mouse egg activation. PKCalpha and gamma accumulate in the egg cortex 25 min post-activation, while only PKCalpha accumulates at the contractile ring of the forming second polar body about 1.5 h post-activation. PKCzeta exhibits some unique features that resulted in it being the focus of more extensive analysis. PKCzeta is tightly associated with the meiotic spindle as determined by detergent extraction and is closely associated with alpha-tubulin as determined by FRET analysis in the metaphase II (MII) egg. In addition, after egg activation, PKCzeta remains associated with the spindle as it transits into anaphase II and later telophase II, becoming associated with the midzone microtubules. Antibodies to the active form of PKCzeta are enriched on the spindle poles and later in development on the midzone microtubules. Active PKCzeta also is enriched in both pronuclei in the 6-h post-fertilization and in the 14-h post-fertilization embryo as well as in the nuclei of the two-cell embryo. Inhibition of PKCzeta, but not inhibition of other isotypes of PKC, results in rapid disruption of the meiotic spindle. This study suggests that PKCzeta has a role in spindle stability, while other PKC isotypes have different roles in the conversion of the egg to the zygote. PMID- 15355788 TI - Redistribution of the kinesin-II subunit KAP from cilia to nuclei during the mitotic and ciliogenic cycles in sea urchin embryos. AB - KAP is the non-motor subunit of the heteromeric plus-end directed microtubule (MT) motor protein kinesin-II essential for normal cilia formation. Studies in Chlamydomonas have demonstrated that kinesin-II drives the anterograde intraflagellar transport (IFT) of protein complexes along ciliary axonemes. We used a green fluorescent protein (GFP) chimera of KAP, KAP-GFP, to monitor movements of this kinesin-II subunit in cells of sea urchin blastulae where cilia are retracted and rebuilt with each mitosis. As expected if involved in IFT, KAP GFP localized to apical cytoplasm, basal bodies, and cilia and became concentrated on basal bodies of newly forming cilia. Surprisingly, after ciliary retraction early in mitosis, KAP-GFP moved into nuclei before nuclear envelope breakdown, was again present in nuclei after nuclear envelope reformation, and only decreased in nuclei as ciliogenesis reinitiated. Nuclear transport of KAP GFP could be due to a putative nuclear localization signal and nuclear export signals identified in the sea urchin KAP primary sequence. Our observation of a protein involved in IFT being imported into the nucleus after ciliary retraction and again after nuclear envelope reformation suggests KAP115 may serve as a signal to the nucleus to reinitiate cilia formation during sea urchin development. PMID- 15355789 TI - The Sine oculis/Six class family of homeobox genes in jellyfish with and without eyes: development and eye regeneration. AB - The development of visual organs is regulated in Bilateria by a network of genes where members of the Six and Pax gene families play a central role. To investigate the molecular aspects of eye evolution, we analyzed the structure and expression patterns of cognate members of the Six family genes in jellyfish (Cnidaria, Hydrozoa), representatives of a basal, non-bilaterian phylum where complex lens eyes with spherical lens, an epidermal cornea, and a retina appear for the first time in evolution. In the jellyfish Cladonema radiatum, a species with well-developed lens eyes in the tentacle bulbs, Six1/2-Cr and Six3/6-Cr, are expressed in the eye cup. Six4/5-Cr is mainly expressed in the manubrium, the feeding, and sex organ. All three Six genes are expressed in different subsets of epidermal nerve cells, possibly of the RFamide type which are part of a net connecting the different eyes with each other and the effector organs. Furthermore, expression is found in other tissues, notably in the striated muscle. During eye regeneration, expression of Six1/2-Cr and Six3/6-Cr is upregulated, but not of Six4/5-Cr. In Podocoryne carnea, a jellyfish without eyes, Six1/2-Pc and Six3/6-Pc are also expressed in the tentacle bulbs, Six1/2-Pc additionally in the manubrium and striated muscle, and Six3/6-Pc in the mechanosensory nematocytes of the tentacle. The conserved gene structure and expression patterns of all Cladonema Six genes suggest broad conservation of upstream regulatory mechanisms in eye development. PMID- 15355790 TI - string(cdc25) and cyclin E are required for patterned histone expression at different stages of Drosophila embryonic development. AB - Metazoan replication-dependent histone mRNAs accumulate to high levels during S phase as a result of an increase in the rate of histone gene transcription, pre mRNA processing, and mRNA stability at the G1-S transition. However, relatively little is known about the contribution of these processes to histone expression in the cell cycles of early development, which often lack a G1 phase. In post blastoderm Drosophila embryos, zygotic expression of the stg(cdc25) phosphatase in G2 activates cyclin/cdc2 kinases and triggers mitosis. Here we show that histone transcription initiates in late G2 of cycle 14 in response to stg(cdc25) and in anticipation of S phase of the next cycle, which occurs immediately following mitosis. Mutation of stg(cdc25) arrests cells in G2 and prevents histone transcription. Expression of a mutant form of Cdc2 that bypasses the requirement for stg(cdc25) activates histone transcription during G2 in stg(cdc25) mutant embryos. Thus, in these embryonic cycles, histone transcription is controlled by the principal G2-M regulators, string(cdc25), and cdc2 kinase, rather than solely by regulators of the G1-S transition. After the introduction of G1-S control midway through embryogenesis, histone expression depends on DNA replication and the function of cyclin E, and no longer requires stg(cdc25). Thus, during the altered cell cycles of early animal development, different cell cycle mechanisms are employed to ensure that the production of histones accompanies DNA synthesis. PMID- 15355791 TI - Cell cycle-coupled [Ca(2+)](i) oscillations in mouse zygotes and function of the inositol 1,4,5-trisphosphate receptor-1. AB - Sperm entry in mammalian eggs initiates oscillations in the concentration of free calcium ([Ca(2+)](i)). In mouse eggs, oscillations start at metaphase II (MII) and conclude as the zygotes progress into interphase and commence pronuclear (PN) formation. The inositol 1,4,5-trisphosphate receptor (IP(3)R-1), which underlies the oscillations, undergoes degradation during this transition, suggesting that one or more of the eggs' Ca(2+)-releasing machinery components may be regulated in a cell cycle-dependent manner, thereby coordinating [Ca(2+)](i) responses with the cell cycle. To ascertain the site(s) of interaction, we initiated oscillations at different stages of the cell cycle in zygotes with different IP(3)R-1 mass. In addition to sperm, we used two other agonists: porcine sperm factor (pSF), which stimulates production of IP(3), and adenophostin A, a non hydrolyzable analogue of IP(3). None of the agonists tested induced oscillations at interphase, suggesting that neither decreased IP(3)R-1 mass nor lack of production or excessive IP(3) degradation can account for the insensitivity to IP(3) at this stage. Moreover, the releasable Ca(2+) content of the stores did not change by interphase, but it did decrease by first mitosis. More importantly, experiments revealed that IP(3)R-1 sensitivity and possibly IP(3) binding were altered at interphase, and our data demonstrate stage-specific IP(3)R-1 phosphorylation by M-phase kinases. Accordingly, increasing the activity of M phase kinases restored the oscillatory-permissive state in zygotes. We therefore propose that the restriction of oscillations in mouse zygotes to the metaphase stage may be coordinated at the level of IP(3)R-1 and that this involves cell cycle stage-specific receptor phosphorylation. PMID- 15355792 TI - Protein kinase CK2 is required for dorsal axis formation in Xenopus embryos. AB - Dorsal axis formation in Xenopus embryos is dependent upon asymmetrical localization of beta-catenin, a transducer of the canonical Wnt signaling pathway. Recent biochemical experiments have implicated protein kinase CK2 as a regulator of members of the Wnt pathway including beta-catenin. Here, we have examined the role of CK2 in dorsal axis formation. CK2 was present in the developing embryo at an appropriate time and place to participate in dorsal axis formation. Overexpression of mRNA encoding CK2 in ventral blastomeres was sufficient to induce a complete ectopic axis, mimicking Wnt signaling. A kinase inactive mutant of CK2alpha was able to block ectopic axis formation induced by XWnt8 and beta-catenin and was capable of suppressing endogenous axis formation when overexpressed dorsally. Taken together, these studies demonstrate that CK2 is a bona fide member of the Wnt pathway and has a critical role in the establishment of the dorsal embryonic axis. PMID- 15355793 TI - eor-1 and eor-2 are required for cell-specific apoptotic death in C. elegans. AB - Programmed cell death occurs in every multicellular organism and in diverse cell types yet the genetic controls that define which cells will live and which will die remain poorly understood. During development of the nematode Caenorhabditis elegans, the coordinated activity of four gene products, EGL-1, CED-9, CED-4 and CED-3, results in the death of essentially all cells fated to die. To identify novel upstream components of the cell death pathway, we performed a genetic screen for mutations that abolish the death of the hermaphrodite-specific neurons (HSNs), a homologous pair of cells required for egg-laying in the hermaphrodite. We identified and cloned the genes, eor-1 and eor-2, which are required to specify the fate of cell death in male HSNs. In addition to defects in HSN death, mutation of either gene leads to defects in coordinated movement, neuronal migration, male tail development, and viability; all consistent with abnormal neuronal differentiation. eor-1 encodes a putative transcription factor related to the human oncogene PLZF. eor-2 encodes a novel but conserved protein. We propose that eor-1 and eor-2 function together throughout the nervous system to promote terminal differentiation of neurons and function specifically in male HSNs to promote apoptotic death of the HSNs. PMID- 15355795 TI - Phenotypic and functional characteristics of spermatogonial stem cells in rats. AB - Spermatogonial stem cells (SSCs) are at the foundation of the highly productive spermatogenic process that continuously produces male gametes throughout postnatal life. However, experimental evaluation of SSCs in postnatal testes is complicated because these cells are extremely rare and few defining morphology or biochemical characteristics are known. In this study, we used the spermatogonial transplantation functional assay, combined with fluorescence-activated cell sorting (FACS) analysis to identify cellular, biochemical and surface antigenic characteristics of SSCs in rat testes during development. Our results demonstrated that forward scatter (FSc)(hi), side scatter (SSc)(hi), mitochondria membrane potential (DeltaPsim)(lo), Ep-CAM(+), Thy-1(+), beta3-integrin(+) stem cells in neonate rat testes become SSc(lo), DeltaPsim(hi), Ep-CAM(+), Thy-1(lo), beta3-integrin(-) stem cells in pup rat testes. Furthermore, prospective identification of rat testis cell populations (Ep-CAM(+)), highly enriched for SSCs (1 in 13 for neonate; 1 in 8.5 for pup) enabled us to predict the Thy-1 and beta3-integrin status of stem cells in neonate and pup testes, which was subsequently confirmed by transplantation analyses. Systematic characterization of SSCs enabled the production of testis cell populations highly enriched (up to 120-fold) for SSCs and will facilitate future investigations of functional and genomic characteristics. PMID- 15355794 TI - Fgf signaling is required for zebrafish tooth development. AB - We have investigated fibroblast growth factor (FGF) signaling during the development of the zebrafish pharyngeal dentition with the goal of uncovering novel roles for FGFs in tooth development as well as phylogenetic and topographic diversity in the tooth developmental pathway. We found that the tooth-related expression of several zebrafish genes is similar to that of their mouse orthologs, including both epithelial and mesenchymal markers. Additionally, significant differences in gene expression between zebrafish and mouse teeth are indicated by the apparent lack of fgf8 and pax9 expression in zebrafish tooth germs. FGF receptor inhibition with SU5402 at 32 h blocked dental epithelial morphogenesis and tooth mineralization. While the pharyngeal epithelium remained intact as judged by normal pitx2 expression, not only was the mesenchymal expression of lhx6 and lhx7 eliminated as expected from mouse studies, but the epithelial expression of dlx2a, dlx2b, fgf3, and fgf4 was as well. This latter result provides novel evidence that the dental epithelium is a target of FGF signaling. However, the failure of SU5402 to block localized expression of pitx2 suggests that the earliest steps of tooth initiation are FGF-independent. Investigations of specific FGF ligands with morpholino antisense oligonucleotides revealed only a mild tooth shape phenotype following fgf4 knockdown, while fgf8 inhibition revealed only a subtle down-regulation of dental dlx2b expression with no apparent effect on tooth morphology. Our results suggest redundant FGF signals target the dental epithelium and together are required for dental morphogenesis. Further work will be required to elucidate the nature of these signals, particularly with respect to their origins and whether they act through the mesenchyme. PMID- 15355796 TI - Regionalization of cell fates and cell movement in the endoderm of the mouse gastrula and the impact of loss of Lhx1(Lim1) function. AB - Investigation of the developmental fates of cells in the endodermal layer of the early bud stage mouse embryo revealed a regionalized pattern of distribution of the progenitor cells of the yolk sac endoderm and the embryonic gut. By tracing the site of origin of cells that are allocated to specific regions of the embryonic gut, it was found that by late gastrulation, the respective endodermal progenitors are already spatially organized in anticipation of the prospective mediolateral and anterior-posterior destinations. The fate-mapping data further showed that the endoderm in the embryonic compartment of the early bud stage gastrula still contains cells that will colonize the anterior and lateral parts of the extraembryonic yolk sac. In the Lhx1(Lim1)-null mutant embryo, the progenitors of the embryonic gut are confined to the posterior part of the endoderm. In particular, the prospective anterior endoderm was sequestered to a much smaller distal domain, suggesting that there may be fewer progenitor cells for the anterior gut that is poorly formed in the mutant embryo. The deficiency of gut endoderm is not caused by any restriction in endodermal potency of the mutant epiblast cells but more likely the inadequate allocation of the definitive endoderm. The inefficient movement of the anterior endoderm, and the abnormal differentiation highlighted by the lack of Sox17 and Foxa2 expression, may underpin the malformation of the head of Lhx1 mutant embryos. PMID- 15355797 TI - Hierarchy revealed in the specification of three skeletal fates by Sox9 and Runx2. AB - Across vertebrates, there are three principal skeletal tissues: bone, persistent cartilage, and replacement cartilage. Although each tissue has a different evolutionary history and functional morphology, they also share many features. For example, they function as structural supports, they are comprised of cells embedded in collagen-rich extracellular matrix, and they derive from a common embryonic stem cell, the osteochondroprogenitor. Occasionally, homologous skeletal elements can change tissue type through phylogeny. Together, these observations raise the possibility that skeletal tissue identity is determined by a shared set of genes. Here, we show that misexpression of either Sox9 or Runx2 can substitute bone with replacement cartilage or can convert persistent cartilage into replacement cartilage and vice versa. Our data also suggest that these transcription factors function in a molecular hierarchy in which chondrogenic factors dominate. We propose a binary molecular code that determines whether skeletal tissues form as bone, persistent cartilage, or replacement cartilage. Finally, these data provide insights into the roles that master regulatory genes play during evolutionary change of the vertebrate skeleton. PMID- 15355798 TI - Phospholipase Cepsilon regulates ovulation in Caenorhabditis elegans. AB - Phospholipase Cepsilon (PLCepsilon) is a novel class of phosphoinositide-specific PLC with unknown physiological functions. Here, we present the first genetic analysis of PLCepsilon in an intact organism, the nematode Caenorhabditis elegans. Ovulation in C. elegans is dependent on an inositol 1,4,5-trisphosphate (IP(3)) signaling pathway activated by the receptor tyrosine kinase LET-23. We generated deletion mutants of the gene, plc-1, encoding C. elegans PLCepsilon. We observed a novel ovulation phenotype whereby oocytes are trapped in the spermatheca due to delayed dilation of the spermatheca-uterine valve. The expression of plc-1 in the adult spermatheca is consistent with its involvement in regulation of ovulation. On the other hand, we failed to observe genetic interaction of plc-1 with let-23-mediated IP(3) signaling pathway genes, suggesting a complex mechanism for control of ovulation. PMID- 15355799 TI - Three distinct lineages of mesenchymal cells in Ciona intestinalis embryos demonstrated by specific gene expression. AB - The ascidian embryonic mesenchyme, comprising about 900 cells, forms mesodermal tissues or organs of the adult body after metamorphosis. The mesenchyme originates from the A7.6 [trunk lateral cells (TLCs)], B7.7, and B8.5 blastomeres of the 110-cell stage embryo. Previous studies showed that FGF9/16/20 is required for specification of the mesenchyme in Ciona embryos and that two different (A7.6 and B8.5/B7.7) but partially overlapping molecular mechanisms are associated with the expression of a basic helix-loop-helix (bHLH) transcription factor gene, Twist-like1, in the mesenchymal precursors, which triggers the differentiation process of mesenchyme cells. In the present study, we examined whether the three embryonic lineages express the same mesenchyme-specific structural genes under the control of a common mechanism or whether the three lineages are characterized by the expression of genes specific to each of the lineages. We characterized nine mesenchyme-specific genes in Ciona embryos and found that five were expressed in A7.6/B8.5/B7.7, two in B8.5/B7.7, and two in B7.7 only. FGF9/16/20 and Twist-like1 were required for the expression of all the mesenchyme-specific genes, except for three A7.6/B8.5/B7.7-specific genes in A7.6 progenitors. Overexpression of FGF9/16/20 or Twist-like1 upregulated the expression of A7.6/B8.5/B7.7- and B8.5/B7.7-specific genes, while it downregulated the expression of B7.7-specific genes. These results provide evidence that the differentiation of each of the three mesenchyme lineages of Ciona embryos is characterized by the expression of a specific set of genes, whose expression is controlled differentially. PMID- 15355800 TI - [Pharmacogenetics or the promise of a personalized medicine: variability in drug metabolism and transport]. AB - There is much variability in the manner individuals respond to drugs, such that the management of some drugs is problematic. In France, the incidence of hospital admissions related to adverse drug reactions is estimated to be 3.2 %, at an annual cost of over 300 millions euros. Genetic factors affecting the pharmacokinetics and pharmacodynamics of drugs partly explain interindividual variability in drug response. Pharmacogenetic focuses on the molecular mechanisms involved in drug response, and its ultimate goal is the optimisation of drug treatments, both in terms of efficacy and safety. Numerous polymorphisms in genes encoding drug-metabolising enzymes, transporters and receptors have been described and their consequences on disposition and effect of a substantial number of medications have been elucidated. This review focuses on variability of drug metabolism and transport to define the objectives of pharmacogenetics, the molecular bases of interindividual variation in drug response and the methods used for the evaluation of the individual risk of drug failure or toxicity. Some clinical applications of pharmacogenetics have already been developed in routine medicine resulting in significant improvement in patient treatment. The clinical validation of an increasing number of pharmacogenetic tests, as well as the development of new highly efficient technologies for genotyping (real-time PCR, DNA chips) should further promote pharmacogenetics in clinical practice and lead to the development of a patient-tailored drug therapy. PMID- 15355801 TI - [Diagnosis of myeloid hematologic malignancies: contributions of the 2001 World Health Organization (WHO) classification]. AB - Although the French-American-British (FAB) morphologic classification of myeloid malignancies has been accepted for many years, the important advances in cytogenetic, immunophenotype and genetic fields needed to be integrated in an updated approach using all the available informations. Thus, the World Health Organization (WHO) Classification of haematopoietic malignancies not only incorporates morphology, immunophenotype, cytogenetic and molecular features, but also ensures to be clinically useful. This collaborative project has begun in 1995 and has been published in 2001. The proposed WHO classification is less a disruption with regard to the FAB classification than an updated revision where morphology is always important. These review emphasises the modifications proposed in the new WHO classification concerning the myeloid disorders. PMID- 15355802 TI - [Cryoglobulinemia]. AB - Cryoglobulins are immunoglobulins or immune complexes which precipitate at a temperature lower than 37 degrees C and redissolve when rewarmed. Cryoglobulins can be asymptomatic. When not, clinical features are in most cases cutaneous, renal or neurological. Cryoglobulinemia are associated with malignant haematological disorder, connective tissue disease or infection process especially infection with hepatitis C virus. In some case, no aetiology can be found, in theses cases cryoglobulinemia are called primary or essential cryoglobulinemia. For the last ten years it has been widely demonstrated that virus C infection causes the most part of essential cryoglobulinemia. PMID- 15355804 TI - [Variations according to age and gender for creatinine clearance estimated with the Cockroft and Gault formula in a selected population of ambulatory adults]. AB - The distribution and variations according to age and gender for the creatinine clearance calculated by the Cockcroft and Gault formula (Cl C-G) were determined thanks to a case-control study on a population of selected adults with a plasma creatinine concentration < 137 micromol/L (man) or < 104 micromol/L (woman) and a plasma urea concentration < 8.3 mmol/L. 286 men and 270 women aged between 18 and 80 were considered. The frequency distributions for creatinine (dosed by a kinetic Jaffe method ) and Cl C-G are Gaussian (p < 0.01; Kolmogorov-Smirnov test). Cl C-G diminishes with age (r = 0.73; p < 0.001), average: 10 mL/min/decade (man) and 11 mL/min/decade (woman). No significant relation could be found between the weight/creatininaemia ratio and age. Cl C-G is weaker for women: 11 to 18 mL/min according to age. 53% of women older than 70 years have an estimated glomerular filtration rate below 60 mL/min. The 2.5th centiles for Cl C G values vary, for man: from 105 mL/min (confidence interval 95%: 100-110 mL/min) between 18 and 30 years old, to 53 mL/min (49-57) between 71 and 80 years old; for woman: from 85 mL/min (80-90) between 18 and 30 years old to 42 mL/min (38 46) between 71 and 80 years old. With such results, the correcting factor related to Cockcroft and Gault's formula gender seems to be insufficient. New reference values and limits of decision of Cl C-G must be determined for each analytical method. PMID- 15355803 TI - [Rape drugs: pharmalogical and analytical aspects]. AB - Rape drugs or compounds used for chemical submission are current hot topics of numerous media based on a few well-documented identified cases. In the aim of considering the compounds potentially involved and subsequently the samples to collect and the toxicological analyses to perform, and according to the aggressor's viewpoint (victim submission and impunity of himself or herself), the characteristics of such compounds were drawn following the drug pharmacological properties. The compounds or therapeutic classes potentially used are numerous and diverse because the expected effects can be obtained by many neuropharmacological mechanisms or combinations of mechanisms. However, a few drugs (i.e. several benzodiazepines, and gamma-hydroxybutyrate) seem to be the ideal candidates owing to advantageous pharmacological properties (low blood concentrations, short elimination half-life) and practical ones (availability, galenic forms). It appears that the quality and precocity of biological specimen collection, the use of specific and sensitive analytical techniques, and the collaboration between the clinician and the toxicologist, are the essential keys for successful toxicological investigations when a case of chemical submission is suspected. PMID- 15355805 TI - [Cholesterol ester fatty acid composition in Tunisian type 2 diabetics with and without cardiovascular complications]. AB - BACKGROUND: Lipid and glycemic imbalances are frequent disorders found in diabetes type 2. These disorders are influenced by dietary means. AIM: to investigate saturated fatty acids (SFA), polyunsaturated fatty acids (PUFS) and oleic acid of cholesterol ester fraction in non-insulin dependant diabetes mellitus without cardiovascular complications (NIDDM), non-insulin dependant diabetes mellitus with cardiovascular complications (NIDDMc) and healthy controls. METHODS: The composition of cholesterol ester fatty acids in 35 NIDDM, 33 NIDDMc and 32 controls were measured by gas-chromatography. Glycaemia and lipid profile were measured using commercial kits. RESULTS: Compared to NIDDM and to controls, NIDDMc showed a significant increase of different SFA (C12:0, C14:0, C16:0, C18:0). Oleic acid (C18:1) was significantly decreased in NIDDMc and NIDDM compared to controls (15,88 +/- 2,34 and 22,66 +/- 4,14 vs 28,18 +/- 2,90). Linoleic acid (C18:2) was significantly increased in NIDDMc compared to NIDDM and controls (52,59 +/- 5,50 vs 49,29 +/- 8,58 and 39,26 +/- 10,46). Linolenic acid (C18:3) and arachidonic acid (C20:4) were significantly decreased in NIDDMc compared to NIDDM and to controls. Linoleic acid (C18:2) / linolenic acid (C18:3) ratio was increased in NIDDMc. CONCLUSION: Linoleic (C18:2) acid excess intake found in our NIDDMc could emphasize arachidonic synthesis which is directly transformed while an inflammatory syndrome observed in coronary pathologies. PMID- 15355806 TI - [A young woman with severe hyponatremia]. AB - We report here the case of a young woman, who came by herself at the emergency department, presenting with a severe hyponatremia (106 mmol/L), as shown by her initial biological report. The biological comment leads us to review the hyponatremia considering on one hand osmolarity and on the other hand volemia. Patient's tests results showed hyponatremia with hypoosmolarity and isovolemia, due to her potomania. At last, the authors expose the main guidelines for the correction of hyponatremia. PMID- 15355807 TI - [Automated kinetic assay of plasmatic L-asparaginase activity undergoing therapy for acute lymphoblastic leukemia]. AB - The L-asparaginase is a critical drug for the treatment of acute lymphoblastic leukaemia, that achieves blood L-asparagin depletion. However, such a therapy is associated with a high rate of negative side effects, particularly antibody synthesis against L-asparaginase. This therefore decreases therapy efficiency requiring the monitoring of L-asparaginase activity since L-asparagin determination is not easy. We compared here the results obtained with an automated kinetic enzymatic method to those obtained with the most commonly used Nessler reagent method. The correlation coefficient, r = 0,992, obtained was very good, and the allometric regression line was y = 1,038x - 0,37 microkat/L. We also showed that the specificity and the precision were better with the enzymatic method than the Nessler one. Moreover, the enzymatic method was easier and required less time to perform. Finally, the method appears able to perform real time monitoring of the therapy. PMID- 15355808 TI - [Fast diagnostic tests in the management of group A beta-heamolytic streptococcal pharyngitis]. AB - OBJECTIVE: Routine clinical diagnosis of Streptococcus pyogenes in pharyngitis is not always easy. The use in common practice of rapid diagnosis test (RDT), might offer a best control of the antibiotic treatments. The aim of this study is to present seven rapid diagnosis tests, to assess their feasibility and finally to determine the bacteriological correlation. METHOD: We propose to compare the results obtained with seven RDT, and to assess their interest in medical diagnosis for group A streptococcus pharyngitis. A prospective study was conducted for three months, a RDT was performed for children (n=75) between eight and fourteen years old presenting acute pharyngitis. Several throat sampling were performed to order cultures. RESULTS: The group A streptococcus was isolated in 33% (n=25) of throat sampling. Comparing cultures results, and for all studied tests, we obtained comparable performances with manufacturer data, specificity upper than 94% and sensitivity upper than 88%. CONCLUSION: All assessed RDT may offer to physicians a decision-making tool for rapid diagnosis. However, because of its complexity, the agglutination test can be used only in pathology laboratories. PMID- 15355809 TI - [Report of a case of Lyell syndrome]. AB - Toxic epidermal necrolysis (TEN) is a rare drug-induced disease characterized by extensive epidermal destruction. We reported here a case of Lyell syndrome which happened few hours later after treatment associating lincomycine chlorhydrate with nonsteroidol anti-inflammatory drugs. The 28-year-old female patient developed many visceral complications with biochemical and haematological disorders. This syndrome is a dermatological emergency whose vital prognosis is displayed. PMID- 15355810 TI - [Should blood samples be drawn on heparin-anticoagulant for lipid analysis?]. AB - Lipid profile is often performed on heparinized-plasma because nothing in particular is explained in the technical data sheet about anticoagulant and because few data (mainly with EDTA anticoagulant) are available in literature. In order to evaluate heparinized-plasma vs serum differences, 50 normo- or hyperlipidemic samples were collected and assayed in 3 clinical laboratories in Lyon on Hitachi analysers with Roche Diagnostic reagents. Lipid values are lower in plasma than in serum; the average negative bias for cholesterol levels is 2 to 4,5% and for triglycerides about 3%, depending on the laboratory; the effect on HDL-cholesterol values is not significant. These results were confirmed by manual procedure on 28 samples with Roche Diagnostic reagent and two similar other reagents (Biomerieux and Randox). A negative bias of 4% on total cholesterol and triglycerides levels is not very important for clinical diagnosis but it is more serious for LDL-cholesterol estimated with Friedewald equation; LDL-cholesterol value needs accuracy because it is a therapeutic goal with statin therapy and an high negative bias (until 0,70 g/L in our results) is unacceptable; moreover, there is a real risk of providing false total cholesterol results. Therefore it is essential to collect blood for lipid profile without any anticoagulant. PMID- 15355811 TI - [Campylobacter fetus subsp fetus bacteremia in a female patient infected by the hepatitis C virus]. AB - Campylobacter fetus subsp fetus was identified as an unusual etiologic agent of septicemia in an immuno-compromized patient VHC positive by utilizing a 16S rRNA molecular kit in our hospital's clinical laboratory. This method would appear as a performing approach to identify pathogens when discrepancies exist between phenotypical tests. PMID- 15355812 TI - [Anti-GD1b IgG positive case of overlapping Ficher's and Guillain-Barre syndromes]. AB - We describe a patient who developed overlapping sensory ataxic form of Guillain Barre syndrome (GBS) and Miller Fisher syndrome (MFS) following Campylobacter jejuni infection. Two cerebrospinal fluid examinations shown albuminocytologic dissociation associated with Campylobacter jejuni infection after tongue pierced. He had high titers of monospecific anti-GD1b IgG antibody. Because of the rarety of this disorder the diagnostic was difficult. There is a close association of IgG anti-ganglioside GD1b antibodies in sensory ataxic GBS. The findings of the present study show that antibody to GD1b ganglioside is one of the immunological factors in the pathogenesis of sensory ataxic form of GBS, a rare specific immuno clinical variant form of GBS with prominent sensory ataxia. PMID- 15355813 TI - [Unexpected occurence of hemoglobin Athens-Georgia]. AB - The following report concerned a 47 year old Caucasian diabetic patient. Routine HPLC of HbA1c (Variant II Biorad Laboratories - hemoglobin A1c program) resulted only in the evidence of HbF (1%) and increase in HbA1c (10%). Considering the presence of HbF a standard agarose gel electrophoresis of patient's hemoglobin was performed and revealed the presence of Hb Athens-Georgia. Consequently the occurrence of HbF during determination of HbA1c by HPLC should lead to perform a standard hemoglobin electrophoresis in order to explore an hidden, unsuspected and clinically silent occurrence of rare Hb variant or additional unsuspected increase in HbA2. PMID- 15355814 TI - [A case of celiac disease with late diagnosis by very long prothrombin and activated partial prothrombin times]. AB - Coeliac disease is usually revealed by intestinal symptoms, but less frequently by deficiency symptoms. Early screening is very important to avoid with appropriate diet an intestinal lymphoma or epidermoid cancer. We report here the case of a 68-year old woman where coeliac disease was pointed out by very long Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT). Clinical examination was strictly normal except for leanness, a small height, and several diarrhoea episodes 3 or 4 times a year. Other blood tests showed a macrocytic anemia, a fibrinogen level slightly above the upper limit, a decreased proteinaemia and albuminaemia, and a sideraemia at the lower normal limit. Liver tests pointed to a cytolysis. Vitamin K-dependent factors were decreased. A perfusion of vitamin-K allowed getting a normal PT. Duodenofiberscopy with biopsy allowed the diagnosis of coeliac disease. Neither lymphoma nor epidermoid cancer were detected. A gluten-free diet allowed the disappearing of digestive symptoms, weight rising and return to a normal PT. Searching for a coeliac disease is therefore relevant in aged patients even when very faint clinical or biological symptoms of malabsorption appear, particularly when PT is longer than the control with decreased vitamin-K dependant factors. PMID- 15355815 TI - [Stability of blood glucose collected with or without antiglycolytic agent]. AB - Glycolysis in blood samples is well-known to induce a rapid decrease of glucose concentration and the use of an antiglycolysis is supposed to prevent this phenomenon. Fifteen healthy volunteers were included in this study. The aim was to evaluate the stability of glucose concentration in regard to the type of blood collection tube used (with or without antiglycolytic agent (monoiodoacetate)) and the time before centrifugation. During the first two hours, a similar decrease was observed with both kind of tube (about 9% in two hours), then the effect of the antiglycolytic agent became significant. It is re-emphasised that glycolysis inhibitor should be used for glucose determination, especially when centrifugation is delayed. PMID- 15355816 TI - [Why is this journal read? New printing order of the AKD?]. PMID- 15355817 TI - Some aspects of cardiology practice in Lebanon. PMID- 15355818 TI - [The relationship between serum C-reactive protein levels and coronary artery disease in patients with stable angina pectoris and positive exercise stress test]. AB - OBJECTIVE: In this study we aimed to investigate whether serum C- reactive protein (CRP) levels are related with the incidence of coronary artery disease (CAD) and selection of management approaches in stable angina pectoris (SAP). METHODS: Overall 134 patients (pts) with SAP and positive exercise stress test were investigated. All pts were divided into two groups according to the baseline levels of CRP. In group 1 (mean age 57.8+/-10.3 years) there were 41 pts with the level of CRP >0.50mg/dl (high levels of CRP) and group 2 consisted of 93 pts (mean age 56.0+/-11.7 years) with the CRP levels <0.50mg/dl (normal levels of CRP). We investigated the relationship between CRP levels with coronary artery disease and treatment strategies. RESULTS: There were no significant differences in age, sex, hypertension and hyperlipidemia between groups. In group 1 (n=41) 36 pts, and in group 2 (n=93) 58 pts had CAD (p= 0.004). We found statistically significant relationship between high levels of CRP and smoking and diabetes mellitus. After adjustment of these risk factors by multivariate regression analyses the CRP association with CAD become attenuated but was still statistically significant (p=0.03). CONCLUSION: In this study we found that high level CRP is an independent strong marker of CAD in middle-aged patients with stable angina and positive treadmill exercise test. There was no correlation between CRP levels and interventional procedures. PMID- 15355819 TI - [C-reactive protein and coronary artery disease]. PMID- 15355820 TI - Triple valve surgery: a 25-year experience. AB - OBJECTIVE: Surgical treatment of rheumatic valvular disease still constitutes a significant number of cardiac operations in developing countries. Despite improvements in myocardial protection and cardiopulmonary bypass techniques, triple valve operations (aortic, mitral and tricuspid valves) are still challenging because of longer duration of cardiopulmonary bypass and higher degree of myocardial decompensation. This study was instituted in order to assess results of triple valve surgery. METHODS: Between 1977 and 2002, 34 patients underwent triple valve surgery in our clinic by the same surgeon (EB). Eleven patients underwent triple valve replacement (32.4%) and 23 underwent tricuspid valve annuloplasty with aortic and mitral valve replacements (67.6%). RESULTS: There was no significant difference between the two groups of patients who underwent triple valve replacement and aortic and mitral valve replacement with tricuspid valve annuloplasty. There were 4 hospital deaths (11.8%) occurring within 30 days. The duration of follow-up for 30 survivors ranged from 6 to 202 months (mean 97 months). The actuarial survival rates were 85%, 72%, and 48% at 5, 10, and 15 years respectively. Actuarial freedom from reoperation rates at 5, 10, and 15 years was 86.3%, 71.9%, and 51.2%, respectively. Freedom from cerebral thromboembolism and anticoagulation-related hemorrhage rates, expressed in actuarial terms was 75.9% and 62.9% at 5 and 10 years. Major cerebral complications occurred in 10 of the 30 patients. CONCLUSION: We prefer replacing, if repairing is not possible, the tricuspid valve, with a bileaflet mechanical prosthesis in a patient with valve replacement of the left heart who will be anticoagulated in order to avoid unfavorable properties of bioprosthesis like degeneration and of old generation mechanical prosthesis like thrombosis and poor hemodynamic function. In recent years, results of triple valve surgery either with tricuspid valve conservation or valve replacement in suitable cases have become encouraging with improvements in surgical techniques and myocardial preservation methods. PMID- 15355821 TI - [Serum level of insulin-like growth factor-1 and insulin-like growth factor binding protein-3 in acute coronary syndromes and relationship with prognosis]. AB - OBJECTIVE: The aim of the present study was to examine the levels of insulin-like growth factor (IGF-I) and binding protein-3 (IGFBP-3) in acute coronary syndrome (ACS) and their relationship with prognosis. METHODS: Thirty patients with ACS (22 male, 8 female) were included in our study. Patient's population included 20 patients with ST elevation myocardial infarction (STEMI) and 10 with non-ST elevation ACS. Death, re-infarction, revascularization and malignant arrhythmia were monitored during 3 months. Study group was compared with 20 healthy subjects (Controls). Blood samples were collected in the first 24 hours and at the end of third month. Serum IGF-I and IGFBP-3 levels were determined by radioimmunoassay method. RESULTS: We found decreased level of IGF-I only in the STEMI group (105+/ 84 ng/ml vs. 715+/-150 ng/ml, p<0.0001). There were no significant differences in IGFBP-3 levels between two groups. Serum IGF-I levels were significantly increased after 3rd month in the STEMI group (356+/-72 ng/ml vs. 105+/-84 ng/ml, p=0.025). There was no relationship between IGF-I, IGFBP-3 levels and cardiovascular events occurred during 90 days of follow-up. CONCLUSION: These data allows to suggest that significantly decreased level of IGF-I in STEMI group of ACSs can be used as a marker of myocardial necrosis. There was no relationship between IGF-I level and cardiovascular events occurred in 90 days, so this parameter can not be used as a negative prognostic factor. PMID- 15355822 TI - [The effects of clarithromycin treatment on cardiac events in acute coronary syndrome patients]. AB - OBJECTIVE: The assessment of short duration early clarithromycin treatment on major cardiac events in acute coronary syndrome patients. METHODS: One hundred and thirteen patients with acute coronary syndrome had been enrolled in the study in a prospective manner. Fifty-seven of 113 patients received peroral clarithromycin 1g/day for 14 days in addition to standard therapy. The remaining 56 patients were considered as control group. The treatment and control groups had similar major cardiac risk factors such as diabetes, hypertension, dyslipidemia and smoking habits. The occurrence of unstable angina pectoris, non ST elevation myocardial infarction and ST elevation myocardial infarction was comparable in both groups. The use of thrombolytic therapy and glycoprotein IIb/IIIa receptor blockers administration was also similar in both groups. The patients were followed for major cardiac events for 6 months. RESULTS: During the follow-up, no difference was observed between groups in the occurrence of unstable angina pectoris, myocardial infarction, the need for revascularization with percutaneous coronary intervention or cardiac surgery and cardiac death. We observed a reduction of myocardial infarction and cardiac death occurrence and an increase in the necessity of percutaneous interventions in the treatment group even though this difference did not reach statistical significance. CONCLUSION: No benefit of short duration early clarithromycin therapy was observed in the occurrence of major cardiac events in acute coronary syndromes. Studies with longer treatment and follow-up period using different antibiotics are necessary to elucidate the possible effect of antibiotics on major cardiac events in patients with acute coronary syndrome. PMID- 15355823 TI - [The effect of left ventricular geometry on myocardial performance index in hypertensive patients]. AB - OBJECTIVE: The aim of this study was to investigate the relationship between the myocardial performance index (MPI) and left ventricular (LV) geometry in hypertensive patients. METHODS: The MPI, which is a marker of systolic and diastolic ventricular function, was measured in 64 hypertensive patients and in 15 healthy persons (Control). According to the value of relative wall thickness (RWT) and LV mass index (LVMI), hypertensive patients were subdivided into four groups: normal (N), 17 patients (26.6%); concentric remodeling (CR), 21 patients (32.8%); concentric hypertrophy (CH), 16 patients (25%); and eccentric hypertrophy (EH), 10 patients (15.6%). RESULTS: A higher MPI was found in all patient groups (N, 0.56+/-0.11; CR, 0.59+/-0.11; CH, 0.68+/-0.19; EH, 0.57+/ 0.10) compared with the controls (0.44+/-0.09) (p=0.004, p<0.001, p<0.001 and p = 0.002, respectively). In the CH group, the MPI was also higher than in N, CR and EH groups (p=0.006, p<0.03 and p=0.009, respectively). No significant difference was found among N, CR and EH groups. The MPI was correlated with LVMI (r=0.28, p=0.014), RWT (r=0.24, p=0.035) and interventricular septum diastolic thickness (r=0.32, p=0.004). CONCLUSION: The systolic and diastolic LV functions are impaired in all subgroups of hypertensive patients according to their LV geometry compared to control group. This impairment is more advanced in patients with concentric hypertrophy than in those with the other LV geometric patterns. PMID- 15355824 TI - Prevalence of coronary artery disease in patients undergoing valvular operation due to rheumatic involvement. AB - OBJECTIVE: Rheumatic heart disease is still a major health problem in developing countries. The impact of coronary artery disease (CAD) on or its relation to rheumatic fever is not well established. We aimed to evaluate the prevalence of CAD and atherosclerotic risk factors in patients who underwent valvular surgery due to rheumatic heart disease. METHODS: The records of 346 patients who had undergone rheumatic valvular surgery in a university hospital between 1996 and 2002 were evaluated. RESULTS: Coronary angiography was performed in 218 (63%) patients, of whom 41 (18.8%) had CAD. The mean age of the patients having CAD and normal coronary arteries were 57.3 and 50.5 years respectively (p<0.001). In the study population patients with CAD had significantly increased prevalence of diabetes mellitus (14.6% vs. 4.5%; p=0.02), hypertension (36.6% vs. 16.4%; p=0.003), smoking (51.2% vs. 23.2%; p=0.001) and family history of CAD (39.5% vs. 20.0%; p=0.01) compared to patients with normal coronary arteries. However, the prevalence of dyslipidemia was similar in both groups (45.9% vs. 36.4%; p=0.1). CONCLUSION: These findings suggest that coronary artery disease prevalence in rheumatic valvular disease patients is similar to the normal population of same age. In cases where invasive assessment of valvular lesions is not indicated we suggest coronary angiography to be performed only in patients having clinical suspicion of CAD or multiple risk factors. PMID- 15355825 TI - Assessing the effect of low dose dobutamine on various diastolic function indexes. AB - OBJECTIVE: The aim of this study was to evaluate the effect of low dose dobutamine (LDD) on various diastolic function parameters in patients without wall motion abnormality. METHODS: Thirty-one volunteer patients who had no regional wall motion abnormality were included in the study. Echocardiographic measurements were taken both at pre-dobutamine and during LDD infusion. The peak E velocity, A velocity, the E/A ratio, deceleration time (DT), isovolumetric relaxation time (IVRT), myocardial performance index (MPI) and flow propagation velocity (FPV) were assessed as left ventricular diastolic function parameters. Tissue Doppler velocities were also obtained in order to calculate the E/Em and Em/Am ratios. RESULTS: No significant changes were observed in heart rate, E velocity, A velocity, E/A ratio, E/Em ratio, Em/Am ratio, systolic and diastolic blood pressure with LDD. With LDD, DT (239+/-40 ms vs. 201+/-31 ms, p<0.001), IVRT (109+/-12 ms vs. 94+/-11 ms, p<0.001) and MPI (0.57+/-0.15 vs. 0.44+/-0.22, p<0.001) were found to be decreased, while there was an increase in FPV (45+/-8 cm/s vs. 59+/-10 cm/s, p<0.001) and ejection fraction (64+/-6% vs. 66+/-7%, p<0.05). CONCLUSION: Low dose dobutamine (5 mcg/kg of body weight) improves left ventricular relaxation in patients with normal wall motion, while it has no effect on left ventricular filling pressure index. PMID- 15355826 TI - [The organ effects of systemic inflammation response activated during open heart surgery and current treatment methods]. AB - Multi-organ failure may occur due to activation of systemic inflammatory process with many other factors in open-heart procedures when cardiopulmonary bypass is used. Activation of systemic inflammatory process may cause postoperative complications. Surgical trauma, contact of blood with foreign surface, endotoxemia and ischemia-reperfusion injury are major factors that contribute to activation of inflammatory response. In this review we purposed to investigate the factors which contribute to the systemic inflammatory process, multiorgan dysfunction and the therapeutic modalities during open heart surgery. PMID- 15355827 TI - Lipids, lipoproteins and apolipoproteins among turks, and impact on coronary heart disease. AB - Serum concentrations of lipids, lipoproteins, and apolipoproteins (apo) among Turkish adults have been reviewed in this paper whereby stratification by gender and age groups was provided, together with a description of differences by geographic regions and urban-rural areas. Most of the knowledge was derived from the prospective population-based Turkish Adult Risk Factor (TEKHARF) Study, having already a 13 years' follow-up, but data contributed by the Turkish Heart Study were also outlined. In the setting of a prevalence of metabolic syndrome in 3 out of 8 Turkish adults, Turks have low levels of total cholesterol (mean 185 mg/dl), LDL-cholesterol (mean 116 mg/dl), and HDL-cholesterol (mean 37 and 45 mg/dl in men and women). The latter is associated with comparatively high concentrations of triglycerides (mean 143 mg/dl) and of apo B (mean 115 mg/dl). This suggests that small, dense LDL particles (pattern B) prevail in this population though studies are missing in this regard. In line with this notion are the high levels of total/HDL cholesterol ratio (mean 5.3 in men, 4.5 in women). It is remarkable that women exhibit identical LDL-cholesterol levels as men. The lipid parameter that has changed strikingly since 1990 are the rising triglycerides, accompanying a similar trend in (abdominal) obesity. On multivariate analysis, the best independent lipid predictor of coronary heart disease (CHD) risk among Turks is the TC/HDL-C ratio. A 2-unit increment of TC/HDL-C adds an excess of 68% to both the nonfatal and fatal CHD event risk. When ratios of = or >5.5 in men and = or >5 in women are considered as high risk, slightly more than one-third of Turkish adults, corresponding to 12 million adults, are included by these criteria into high-risk group. A major portion of Turkish adults harbouring total cholesterol concentrations in the 180-200 mg/dl range are at high risk, and we stress the opinion that the upper normal limit of total cholesterol be reduced to 180 mg/dl in Turks, at least in men. PMID- 15355828 TI - [General approach to ventricular arrhythmias]. PMID- 15355829 TI - [Perivalvular pseudoaneurysm, abscess and vegetation along with coronary embolism in aortic prosthetic valve]. PMID- 15355830 TI - Cardiac decompression sickness after hypobaric chamber training: case report of a coronary gas embolism. PMID- 15355831 TI - Cardiac decompression sickness. PMID- 15355832 TI - [Two cases with suspected permanent pacemaker allergy]. PMID- 15355833 TI - Stent implantation to left pulmonary artery stenosis in children: a case report. PMID- 15355834 TI - Double orifice mitral valve and spontaneous echo contrast in the descending aorta. PMID- 15355835 TI - Surgical treatment of the atrial septal aneurysms with concomitant cardiovascular abnormalities. PMID- 15355836 TI - [Endovascular treatment of thoracic aortic aneurysms]. PMID- 15355837 TI - [The role of aerobic exercise after myocardial infarction]. PMID- 15355838 TI - [The effect of glutamate and aspartate on myocardial protection at cardiopulmonary bypass]. PMID- 15355839 TI - The 8th World Congress of Echocardiography and Vascular Ultrasound, Antalya, Turkey, May 6th-9th, 2004. PMID- 15355840 TI - [A case of left main coronary artery aneurysm associated with severe stenosis of left anterior descending artery]. PMID- 15355841 TI - [Catheter-induced left anterior descending coronary artery spasm mimicking atherosclerotic lesion]. PMID- 15355842 TI - From treatment to diagnosis: a huge left ventricular thrombus in a patient with heart failure. PMID- 15355843 TI - Severe myocardial ischemia caused by muscular bridge of the diagonal branch of the left anterior descending coronary artery. PMID- 15355844 TI - [Right atrial mobile thrombus leading to pulmonary embolism]. PMID- 15355845 TI - [Renal cell carcinoma with right atrial invasion]. PMID- 15355846 TI - [Conversation with prof.dr. Remzi Ozcan (interview by prof. Dr. Yilmaz Nisanci)]. PMID- 15355847 TI - Differential expression of divalent metal transporter DMT1 (Slc11a2) in the spermatogenic epithelium of the developing and adult rat testis. AB - Iron is essential for male fertility, and disruptions in iron balance lead to impairment of testicular function. The divalent metal transporter DMT1 is a key modulator of transferrin- and non-transferrin-bound iron homeostasis. As a first step in determining the role of DMT1 in the testis, we have characterized the pattern of DMT1 expression in the developing and adult rat testis. Northern blot analysis and RT-PCR of testis polyadenylated RNA revealed the presence of iron responsive element (IRE) and non-IRE transcripts. Semiquantitative immunoblotting of immature and adult rat testis uncovered the expression of two distinct DMT1 protein species. Immunohistochemistry showed that DMT1 was widespread throughout each seminiferous tubule and was expressed in the intracellular compartment. In the adult rat testis, DMT1 was immunolocalized to both the Sertoli and germ cells. In contrast to the immature testis, expression was dependent on the stage of the spermatogenic cycle. DMT1 was not detected on any plasma membranes in either the developing or the adult testis, suggesting that DMT1 is not primarily responsible for translocating iron across this epithelium. Our data suggest an important role for DMT1 in intracellular iron handling during spermatogenesis and imply that germ cells have a need for a precisely targeted and timed supply of iron. We suggest that DMT1 may, as it does in other tissues, play a role in transporting iron between intracellular compartments and thus may play an important role in male fertility. PMID- 15355848 TI - Cleavage of SNAP-25 and VAMP-2 impairs store-operated Ca2+ entry in mouse pancreatic acinar cells. AB - We recently reported that store-operated Ca(2+) entry (SOCE) in nonexcitable cells is likely to be mediated by a reversible interaction between Ca(2+) channels in the plasma membrane and the endoplasmic reticulum, a mechanism known as "secretion-like coupling." As for secretion, in this model the actin cytoskeleton plays a key regulatory role. In the present study we have explored the involvement of the secretory proteins synaptosome-associated protein (SNAP 25) and vesicle-associated membrane protein (VAMP) in SOCE in pancreatic acinar cells. Cleavage of SNAP-25 and VAMPs by treatment with botulinum toxin A (BoNT A) and tetanus toxin (TeTx), respectively, effectively inhibited amylase secretion stimulated by the physiological agonist CCK-8. BoNT A significantly reduced Ca(2+) entry induced by store depletion using thapsigargin or CCK-8. In addition, treatment with BoNT A once SOCE had been activated reduced Ca(2+) influx, indicating that SNAP-25 is needed for both the activation and maintenance of SOCE in pancreatic acinar cells. VAMP-2 and VAMP-3 are expressed in mouse pancreatic acinar cells. Both proteins associate with the cytoskeleton upon Ca(2+) store depletion, although only VAMP-2 seems to be sensitive to TeTx. Treatment of pancreatic acinar cells with TeTx reduced the activation of SOCE without affecting its maintenance. These findings support a role for SNAP-25 and VAMP-2 in the activation of SOCE in pancreatic acinar cells and show parallels between this process and secretion in a specialized secretory cell type. PMID- 15355849 TI - Polyamine-modulated expression of c-myc plays a critical role in stimulation of normal intestinal epithelial cell proliferation. AB - The nuclear protein c-Myc is a transcription factor involved in the control of cell cycle. Our previous studies indicated that cellular polyamines are absolutely required for cell proliferation in crypts of small intestinal mucosa and that polyamines have the ability to stimulate expression of the c-myc gene. The current study went further to determine whether induced nuclear c-Myc plays a role in stimulation of cell proliferation by polyamines in intestinal crypt cells (IEC-6 line). Exposure of normal quiescent cells after 24-h serum deprivation to 5% dialyzed fetal bovine serum (dFBS) increased both cellular polyamines and expression of the c-myc gene. Increased c-Myc protein formed heterodimers with its binding partner, Max, and specifically bound to the Myc/Max binding site, which was associated with an increase in DNA synthesis. Depletion of cellular polyamines by pretreatment with alpha-difluoromethylornithine (DFMO) prevented increases in c-myc expression and DNA synthesis induced by 5% dFBS. c-Myc gene transcription and cell proliferation decreased in polyamine-deficient cells, whereas the natural polyamine spermidine given together with DFMO maintained c myc gene expression and cell growth at normal levels. Disruption of c-myc expression using specific c-myc antisense oligomers not only inhibited normal cell growth (without DFMO) but also prevented the restoration of cell proliferation by spermidine in polyamine-deficient cells. Ectopic expression of wild-type c-myc by recombinant adenoviral vector containing c-myc cDNA increased cell growth. These results indicate that polyamine-induced nuclear c-Myc interacts with Max, binds to the specific DNA sequence, and plays an important role in stimulation of normal intestinal epithelial cell proliferation. PMID- 15355850 TI - Insulin-induced phosphorylation of ENaC correlates with increased sodium channel function in A6 cells. AB - The purpose of this study was to determine whether there is a correlation between phosphorylation and activity of the epithelial sodium channel (ENaC). The three subunits that form the channel were immunoprecipitated from A6 cells by using specific polyclonal antibodies after labeling cells with (35)S or (32)P. When immune complexes were resolved on SDS-PAGE, the alpha-subunit migrated at 85 and 65 kDa, the beta-subunit at 115 and 100 kDa, and the gamma-subunit at 90 kDa. In the resting state all three subunits were phosphorylated. The alpha-subunit was phosphorylated only in the 65-kDa band, suggesting that the posttranslational modification that gives rise to the rapidly migrating form of alpha is a requirement for phosphorylation. Stimulation with 100 nM insulin for 30 min increased phosphorylation of alpha-, beta-, and gamma-subunits approximately twofold. Exposure to 1 microM aldosterone for 16 h increased protein abundance and phosphorylation proportionately in the three subunits. When insulin was applied to cells pretreated with aldosterone, phosphorylation was also increased approximately twofold, but the total amount of phosphorylated substrate was larger than in control conditions because of the action of aldosterone. This result might explain the synergistic increase in sodium transport under the same conditions. The protein kinase C inhibitor chelerythrine abolished insulin effects and decreased sodium transport and subunit phosphorylation. Together, our findings suggest that ENaC activity is controlled by subunit phosphorylation in cells that endogenously express the channel and the machinery for hormonal stimulation of sodium transport. PMID- 15355851 TI - Activation of chloride currents in murine portal vein smooth muscle cells by membrane depolarization involves intracellular calcium release. AB - The present study describes the first characterization of Ca(2+)-activated Cl(-) currents (I(ClCa)) in single smooth muscle cells from a murine vascular preparation (portal veins). I(ClCa) was recorded using the perforated patch version of the whole cell voltage-clamp technique and was evoked using membrane depolarization. Generation of I(ClCa) relied on Ca(2+) entry through dihydropyridine-sensitive Ca(2+) channels because I(ClCa) was abolished by 1 microM nicardipine and enhanced by raising external Ca(2+) concentration or by application of BAY K 8644. I(ClCa) was characterized by the sensitivity to Cl(-) channel blockers and the effect of altering the external anion on reversal potential. Activation of I(ClCa) after membrane depolarization was dependent on Ca(2+) release from intracellular stores. Thus the amplitude of I(ClCa) was diminished by the SR-ATPase inhibitor cyclopiazonic acid, the inositol 1,4,5 trisphosphate receptor antagonist 2-aminoethoxydiphenyl borate (2-APB), and the ryanodine receptor blocker tetracaine. The degree of inhibition produced by the application of 2-APB and tetracaine together was significantly greater than the effect of each agent applied alone. In current-clamp mode, injection of depolarizing current elicited a biphasic action potential, with the later depolarization being sensitive to niflumic acid (NFA; 10 microM). In isometric tension recordings, NFA inhibited spontaneous contractions. These data support a role for this conductance in portal vein excitability. PMID- 15355852 TI - The role of intracellular pH in cell growth arrest induced by ATP. AB - In this study, we investigated ionic mechanisms involved in growth arrest induced by extracellular ATP in androgen-independent prostate cancer cells. Extracellular ATP reversibly induced a rapid and sustained intracellular pH (pH(i)) decrease from 7.41 to 7.11. Inhibition of Ca(2+) influx, lowering extracellular Ca(2+), and buffering cytoplasmic Ca(2+) inhibited ATP-induced acidification, thereby demonstrating that acidification is a consequence of Ca(2+) entry. We show that ATP induced reuptake of Ca(2+) by the mitochondria and a transient depolarization of the inner mitochondrial membrane. ATP-induced acidification was reduced after the dissipation of the mitochondrial proton gradient by rotenone and carbonyl cyanide p-trifluoromethoxyphenylhydrazone, after inhibition of Ca(2+) uptake into the mitochondria by ruthenium red, and after inhibition of the F(0)F(1)-ATPase with oligomycin. ATP-induced acidification was not induced by either stimulation of the Cl(-)/HCO(3)(-) exchanger or inhibition of the Na(+)/H(+) exchanger. In addition, intracellular acidification, induced by an ammonium prepulse method, reduced the amount of releasable Ca(2+) from the endoplasmic reticulum, assessed by measuring change in cytosolic Ca(2+) induced by thapsigargin or ATP in a Ca(2+)-free medium. This latter finding reveals cross talk between pH(i) and Ca(2+) homeostasis in which the Ca(2+)-induced intracellular acidification can in turn regulate the amount of Ca(2+) that can be released from the endoplasmic reticulum. Furthermore, pH(i) decrease was capable of reducing cell growth. Taken together, our results suggest that ATP-induced acidification in DU-145 cells results from specific effect of mitochondrial function and is one of the major mechanisms leading to growth arrest induced by ATP. PMID- 15355853 TI - Calcium, ATP, and ROS: a mitochondrial love-hate triangle. AB - The mitochondrion is at the core of cellular energy metabolism, being the site of most ATP generation. Calcium is a key regulator of mitochondrial function and acts at several levels within the organelle to stimulate ATP synthesis. However, the dysregulation of mitochondrial Ca(2+) homeostasis is now recognized to play a key role in several pathologies. For example, mitochondrial matrix Ca(2+) overload can lead to enhanced generation of reactive oxygen species, triggering of the permeability transition pore, and cytochrome c release, leading to apoptosis. Despite progress regarding the independent roles of both Ca(2+) and mitochondrial dysfunction in disease, the molecular mechanisms by which Ca(2+) can elicit mitochondrial dysfunction remain elusive. This review highlights the delicate balance between the positive and negative effects of Ca(2+) and the signaling events that perturb this balance. Overall, a "two-hit" hypothesis is developed, in which Ca(2+) plus another pathological stimulus can bring about mitochondrial dysfunction. PMID- 15355854 TI - The molecular basis of skeletal muscle atrophy. AB - Skeletal muscle atrophy attributable to muscular inactivity has significant adverse functional consequences. While the initiating physiological event leading to atrophy seems to be the loss of muscle tension and a good deal of the physiology of muscle atrophy has been characterized, little is known about the triggers or the molecular signaling events underlying this process. Decreases in protein synthesis and increases in protein degradation both have been shown to contribute to muscle protein loss due to disuse, and recent work has delineated elements of both synthetic and proteolytic processes underlying muscle atrophy. It is also becoming evident that interactions among known proteolytic pathways (ubiquitin-proteasome, lysosomal, and calpain) are involved in muscle proteolysis during atrophy. Factors such as TNF-alpha, glucocorticoids, myostatin, and reactive oxygen species can induce muscle protein loss under specified conditions. Also, it is now apparent that the transcription factor NF-kappaB is a key intracellular signal transducer in disuse atrophy. Transcriptional profiles of atrophying muscle show both up- and downregulation of various genes over time, thus providing further evidence that there are multiple concurrent processes involved in muscle atrophy. The purpose of this review is to synthesize our current understanding of the molecular regulation of muscle atrophy. We also discuss how ongoing work should uncover more about the molecular underpinnings of muscle wasting, particularly that due to disuse. PMID- 15355855 TI - Na(+)/H(+) exchanger NHE1 as plasma membrane scaffold in the assembly of signaling complexes. AB - The plasma membrane Na(+)/H(+) exchanger NHE1 has an established function in intracellular pH and cell volume homeostasis by catalyzing electroneutral influx of extracellular Na(+) and efflux of intracellular H(+). A second function of NHE1 as a structural anchor for actin filaments through its direct binding of the ezrin, radixin, and moesin (ERM) family of actin-binding proteins was recently identified. ERM protein binding and actin anchoring by NHE1 are necessary to retain the localization of NHE1 in specialized plasma membrane domains and to promote cytoskeleton-dependent processes, including actin filament bundling and cell-substrate adhesions. This review explores a third function of NHE1, as a plasma membrane scaffold in the assembly of signaling complexes. Through its coordinate functions in H(+) efflux, actin anchoring, and scaffolding, we propose that NHE1 promotes protein interactions and activities, assembles signaling complexes in specialized plasma membrane domains, and coordinates divergent signaling pathways. PMID- 15355856 TI - An osteoclastic protein-tyrosine phosphatase may play a role in differentiation and activity of human monocytic U-937 cell-derived, osteoclast-like cells. AB - This study investigated if an osteoclastic protein-tyrosine phosphatase (PTP), PTP-oc, plays a role in the functional activity and differentiation of osteoclastic cells by determining the effects of overexpression of wild-type (WT) or phosphatase-deficient (PD)-PTP-oc on bone resorption activity and differentiation of human promyelomonocytic U-937 cells, which could be induced to differentiate into "osteoclast-like" cells by phorbol ester/1,25(OH)(2)D(3) treatment. U-937 cells overexpressing WT- or PD-PTP-oc were produced with a transposon-based vector. The size and depth of resorption pits created by WT-PTP oc-overexpressing osteoclast-like cells were greater, while those by PD-PTP-oc overexpressing osteoclast-like cells were less, than those created by control osteoclast-like cells. Overexpression of WT-PTP-oc also enhanced, while overexpression of PD-PTP-oc suppressed, their differentiation into osteoclast like cells. Overexpression of WT-PTP-oc increased apoptosis and proliferation of U-937 cells, and overexpression of PD-PTP-oc reduced cell proliferation. Cells overexpressing WT-PTP-oc has also led to greater c-Src and NF-kappabeta activation, whereas cells overexpressing PD-PTP-oc resulted in less c-Src and NF kappabeta activation. c-Src activation and NF-kappabeta activation each correlated with resorption activity and differentiation into osteoclast-like cells. In summary, these results show that 1) PTP-oc regulates both the activity and the differentiation of osteoclast-like cells derived from U-937 cells; 2) PTP oc enzymatic activity is important to these processes; 3) high PTP-oc enzymatic activity caused an increase in U-937 cell apoptosis and proliferation, leading to no significant changes in the number of viable cells; and 4) some of the PTP-oc actions are mediated in part by the c-Src and/or NF-kappabeta pathways. PMID- 15355857 TI - MKP-1 expression and stabilization and cGK Ialpha prevent diabetes- associated abnormalities in VSMC migration. AB - Diabetes mellitus is a major risk factor in the development of atherosclerosis and cardiovascular disease conditions, involving intimal injury and enhanced vascular smooth muscle cell (VSMC) migration. We report a mechanistic basis for divergences between insulin's inhibitory effects on migration of aortic VSMC from control Wistar Kyoto (WKY) rats versus Goto-Kakizaki (GK) diabetic rats. In normal WKY VSMC, insulin increased MAPK phosphatase-1 (MKP-1) expression as well as MKP-1 phosphorylation, which stabilizes it, and inhibited PDGF-mediated MAPK phosphorylation and cell migration. In contrast, basal migration was elevated in GK diabetic VSMCs, and all of insulin's effects on MKP-1 expression and phosphorylation, MAPK phosphorylation, and PDGF-stimulated migration were markedly inhibited. The critical importance of MKP-1 in insulin inhibition of VSMC migration was evident from several observations. MKP-1 small interfering RNA inhibited MKP-1 expression and abolished insulin inhibition of PDGF-induced VSMC migration. Conversely, adenoviral expression of MKP-1 decreased MAPK phosphorylation and basal migration rate and restored insulin's ability to inhibit PDGF-directed migration in GK diabetic VSMCs. Also, the proteasomal inhibitors lactacystin and MG132 partially restored MKP-1 protein levels in GK diabetic VSMCs and inhibited their migration. Furthermore, GK diabetic aortic VSMCs had reduced cGMP-dependent protein kinase Ialpha (cGK Ialpha) levels as well as insulin-dependent, but not sodium nitroprusside-dependent, stimulation of cGMP. Adenoviral expression of cGK Ialpha enhanced MKP-1 inhibition of MAPK phosphorylation and VSMC migration. We conclude that enhanced VSMC migration in GK diabetic rats is due at least in part to a failure of insulin-stimulated cGMP/cGK Ialpha signaling, MKP-1 expression, and stabilization and thus MAPK inactivation. PMID- 15355859 TI - The tone of pulmonary smooth muscle: ROK and Rho music? PMID- 15355860 TI - Susceptibility to neoplastic and non-neoplastic pulmonary diseases in mice: genetic similarities. AB - Chronic inflammation predisposes toward many types of cancer. Chronic bronchitis and asthma, for example, heighten the risk of lung cancer. Exactly which inflammatory mediators (e.g., oxidant species and growth factors) and lung wound repair processes (e.g., proangiogenic factors) enhance pulmonary neoplastic development is not clear. One approach to uncover the most relevant biochemical and physiological pathways is to identify genes underlying susceptibilities to inflammation and to cancer development at the same anatomic site. Mice develop lung adenocarcinomas similar in histology, molecular characteristics, and histogenesis to this most common human lung cancer subtype. Over two dozen loci, called Pas or pulmonary adenoma susceptibility, Par or pulmonary adenoma resistance, and Sluc or susceptibility to lung cancer genes, regulate differential lung tumor susceptibility among inbred mouse strains as assigned by QTL (quantitative trait locus) mapping. Chromosomal sites that determine responsiveness to proinflammatory pneumotoxicants such as ozone (O3), particulates, and hyperoxia have also been mapped in mice. For example, susceptibility QTLs have been identified on chromosomes 17 and 11 for O3-induced inflammation (Inf1, Inf2), O3-induced acute lung injury (Aliq3, Aliq1), and sulfate-associated particulates. Sites within the human and mouse genomes for asthma and COPD phenotypes have also been delineated. It is of great interest that several susceptibility loci for mouse lung neoplasia also contain susceptibility genes for toxicant-induced lung injury and inflammation and are homologous to several human asthma loci. These QTLs are described herein, candidate genes are suggested within these sites, and experimental evidence that inflammation enhances lung tumor development is provided. PMID- 15355861 TI - ROS to the rescue. PMID- 15355862 TI - Critical period for alveologenesis and early determinants of adult pulmonary disease. PMID- 15355863 TI - Fas/FasL-mediated apoptosis in perinatal murine lungs. AB - Postcanalicular lung development is characterized by a time-specific increase in alveolar epithelial type II cell apoptosis. We have previously demonstrated that, in fetal rabbits, developmental type II cell apoptosis coincides with transient upregulation of the cell death regulator Fas ligand (FasL). The aims of this study were 1) to determine the spatiotemporal patterns of pulmonary apoptosis and Fas/FasL gene expression in the murine model [embryonic day 17 (E17) through postnatal day 5 (P5)], and 2) to investigate the functional involvement of the Fas/FasL system by determining the effect of Fas activation and inhibition on perinatal pulmonary apoptosis. The apoptotic activity of alveolar epithelial type II cells, determined by combined TUNEL labeling and anti-surfactant protein B immunohistochemistry, showed a dramatic increase during the perinatal transition (type II cell apoptotic index <0.1% at E17, 1.5% at P1-P3, and 0.3% at P5). This timing of enhanced type II cell apoptosis coincided with a robust 14-fold increase in Fas mRNA and protein levels and a threefold increase in FasL protein levels; both Fas and FasL immunolocalized to type II and bronchial epithelial cells. In vitro and in vivo exposure of fetal and postnatal murine type II cells to anti-Fas antibody induced a fourfold increase in apoptotic activity that was prevented by administration of a broad-spectrum caspase inhibitor; the pulmonary apoptotic activity of Fas-deficient lpr mice remained unchanged. Conversely, administration of a caspase inhibitor to newborn mice (P1) resulted in marked diminution of pulmonary apoptotic activity. These combined findings strongly implicate the Fas/FasL system as a critical regulator of perinatal type II cell apoptosis. The developmental time dependence of apoptosis-related events in the murine model should facilitate investigations of the regulation of perinatal pulmonary apoptotic gene expression. PMID- 15355864 TI - Plant respiration and elevated atmospheric CO2 concentration: cellular responses and global significance. AB - BACKGROUND: Elevated levels of atmospheric [CO2] are likely to enhance photosynthesis and plant growth, which, in turn, should result in increased specific and whole-plant respiration rates. However, a large body of literature has shown that specific respiration rates of plant tissues are often reduced when plants are exposed to, or grown at, high [CO2] due to direct effects on enzymes and indirect effects derived from changes in the plant's chemical composition. SCOPE: Although measurement artefacts may have affected some of the previously reported effects of CO2 on respiration rates, the direction and magnitude for the effects of elevated [CO2] on plant respiration may largely depend on the vertical scale (from enzymes to ecosystems) at which measurements are taken. In this review, the effects of elevated [CO2] from cells to ecosystems are presented within the context of the enzymatic and physiological controls of plant respiration, the role(s) of non-phosphorylating pathways, and possible effects associated with plant size. CONCLUSIONS: Contrary to what was previously thought, specific respiration rates are generally not reduced when plants are grown at elevated [CO2]. However, whole ecosystem studies show that canopy respiration does not increase proportionally to increases in biomass in response to elevated [CO2], although a larger proportion of respiration takes place in the root system. Fundamental information is still lacking on how respiration and the processes supported by it are physiologically controlled, thereby preventing sound interpretations of what seem to be species-specific responses of respiration to elevated [CO2]. Therefore the role of plant respiration in augmenting the sink capacity of terrestrial ecosystems is still uncertain. PMID- 15355865 TI - The management of extracellular ice by petioles of frost-resistant herbaceous plants. AB - BACKGROUND AND AIMS: Some frost-tolerant herbaceous plants droop and wilt during frost events and recover turgor and posture on thawing. It has long been known that when plant tissues freeze, extracellular ice forms. Distributions of ice and water in frost-frozen and recovered petioles of Trifolium repens and Escholschzia californica were visualized. METHODS: Petioles of intact plants were cryo-fixed, planed to smooth transverse faces, and examined in a cryo-SEM. KEY RESULTS: With frost-freezing, parenchyma tissues shrank to approx. one-third of their natural volume with marked cytorrhysis of the cells, and massive blocks of extracellular icicles grew under the epidermis (poppy) or epidermis and subepidermis (clover), leaving these layers intact but widely separated from the parenchyma except at specially structured anchorages overlying vascular bundles. On thawing, the extracellular ice was reabsorbed by the expanding parenchyma, and surface tissues again contacted the internal tissues at weak junctions (termed faults). These movements of water into and from the fault zones occurred repeatedly at each frost/thaw event, and are interpreted to explain the turgor changes that led to wilting and recovery. Ice accumulations at tri-cellular junctions with intercellular spaces distended these spaces into large cylinders, especially large in clover. Xylem vessels of frozen petioles were nearly all free of gas; in thawed petioles up to 20 % of vessels were gas-filled. CONCLUSIONS: The occurrence of faults and anchorages may be expected to be widespread in frost tolerant herbaceous plants, as a strategy accommodating extracellular ice deposits which prevent intracellular freezing and consequent membrane disruption, as well as preventing gross structural damage to the organs. The developmental processes that lead to this differentiation of separation of sheets of cells firmly cemented at determined regions at their edges, and their physiological consequences, will repay detailed investigation. PMID- 15355866 TI - Sugar-responsive gene expression, invertase activity, and senescence in aborting maize ovaries at low water potentials. AB - BACKGROUND AND AIMS: Ovary abortion can occur in maize (Zea mays) if water deficits lower the water potential (psiw) sufficiently to inhibit photosynthesis around the time of pollination. The abortion decreases kernel number. The present work explored the activity of ovary acid invertases and their genes, together with other genes for sucrose-processing enzymes, when this kind of abortion occurred. Cytological evidence suggested that senescence may have been initiated after 2 or 3 d of low psiw, and the expression of some likely senescence genes was also determined. METHODS: Ovary abortion was assessed at kernel maturity. Acid invertase activities were localized in vivo and in situ. Time courses for mRNA abundance were measured with real time PCR. Sucrose was fed to the stems to vary the sugar flux. KEY RESULTS: Many kernels developed in controls but most aborted when psiw became low. Ovary invertase was active in controls but severely inhibited at low psiw for cell wall-bound forms in vivo and soluble forms in situ. All ovary genes for sucrose processing enzymes were rapidly down-regulated at low psiw except for a gene for invertase inhibitor peptide that appeared to be constitutively expressed. Some ovary genes for senescence were subsequently up regulated (RIP2 and PLD1). In some genes, these regulatory changes were reversed by feeding sucrose to the stems. Abortion was partially prevented by feeding sucrose. CONCLUSIONS: A general response to low psiw in maize ovaries was an early down-regulation of genes for sucrose processing enzymes followed by up regulation of some genes involved in senescence. Because some of these genes were sucrose responsive, the partial prevention of abortion with sucrose feeding may have been caused in part by the differential sugar-responsiveness of these genes. The late up-regulation of senescence genes may have caused the irreversibility of abortion. PMID- 15355867 TI - High levels of genetic diversity throughout the range of the Portuguese wheat landrace 'Barbela'. AB - BACKGROUND AND AIMS: Landrace populations represent an important intra-crop reservoir of biodiversity and source of novel gene alleles for use in breeding programmes. Here the aim was to measure the diversity of a wheat landrace, 'Barbela', from the north of Portugal. METHODS: DNA was extracted from 59 accessions of Barbela collected across its geographical range. Diversity was measured by microsatellite length polymorphisms using 27 primer pairs amplifying 34 polymorphic microsatellite loci. KEY RESULTS: High levels of polymorphism were found, with an average polymorphism information content of 0.52; an average of 4.77 alleles (range 2-11) were present at each locus, and half of these loci showed an additional allele in the reference variety 'Chinese Spring'. CONCLUSIONS: 'Barbela' is maintained from seeds collected by farmers, but it maintains high allelic variation, and no groupings of accessions were detected when analysed by geographical region, farm or climate, indicating that the wheat landrace is a homogeneous entity. The diversity within the farmer-maintained landrace demonstrates the importance of characterization and maintenance of landrace collections before valuable genetic combinations are lost as uniform commercial crops are introduced. PMID- 15355868 TI - Phenotypes in asthma: useful guides for therapy, distinct biological processes, or both? PMID- 15355869 TI - Aspergillosis in "nonimmunocompromised" critically ill patients. PMID- 15355870 TI - Slip of the tongue. PMID- 15355871 TI - Diagnosis and initial management of nonmalignant diseases related to asbestos. PMID- 15355872 TI - Pharyngeal anatomy and severity of obstructive apnea. PMID- 15355873 TI - Neck proprioception and spatial orientation in cervical dystonia. AB - Neck muscle vibration is known to influence body orientation and posture during locomotion and stance in normal subjects. To verify the hypothesis that neck proprioceptive input can be misinterpreted in patients with cervical dystonia (CD), lateral continuous vibration was applied to the sternocleidomastoid muscle during both stepping-in-place and quiet stance, with eyes closed. The orienting responses of CD patients were compared with those of normal subjects. Vibration effects on body orientation during stepping and stance were apparently different from normal, since no effects were seen when all patients' data collapsed were analysed. However, while some patients did not respond to vibratory stimuli regardless of the vibrated side, others had a 'good' side, the stimulation of which produced effects on body orientation similar to those observed in normal subjects. Homogeneous groups within the patient population were identified, based on the vibration-induced responses under stepping conditions. The different orienting or postural responses observed in CD patients were correlated with disease-related features such as spontaneous head position, maximum range of voluntary head yaw, presence or absence of a botulinum toxin treatment and disease duration. Our data suggest that, in CD patients, the reference system used in the control of body orientation in space is either refractory to the lateralized proprioceptive neck input or modified such that the input from both sides produces an orientation shift in the same sense. This would depend on the pathogenesis of the disease or on an adaptive process connected to the head abnormal posture. It seems that this refractoriness spreads to both sides of the neck with the advancement of the disease, thereby possibly entraining a progressive shift from a reference system based on the head to a more reliable egocentric reference. PMID- 15355874 TI - IVIg therapy in brain inflammation: etiology-dependent differential effects on leucocyte recruitment. AB - Several studies have reported beneficial effects of intravenous immunoglobulin (IVIg) in diseases of the neuroaxis. However, IVIg effects on leucocyte recruitment, a hallmark feature of autoimmunity and acute inflammation, remain largely unexplored. Using intravital microscopy, we studied the effects of IVIg on leucocyte recruitment in experimental autoimmune encephalomyelitis, a model of multiple sclerosis. In IVIg-treated mice, a significant decrease in recruitment (rolling and adhesion) was observed prior to and following disease onset, and this was concomitant with improved clinical score. Since much of the recruitment is dependent upon alpha4-integrin (ligand for VCAM-1) we used an in vitro flow chamber system and demonstrated a 60% decrease in alpha4-integrin-dependent leucocyte adhesion to immobilized VCAM-1. Finally, we used leucocytes from multiple sclerosis patients and demonstrated that IVIg treatment decreased recruitment by 60% on human endothelium. However, when we visualized the role of IVIg in a second model of brain inflammation, cerebral ischaemia-reperfusion, IVIg actually promoted the formation of platelet-leucocyte aggregates in post ischaemic cerebral vessels. In conclusion, we report a new mechanism of action of IVIg through interference of alpha4-integrin-dependent leucocyte recruitment in both an animal model and human multiple sclerosis. We also report that IVIg will not be beneficial in all types of pro-adhesive states and may in fact be detrimental in a situation such as stroke. PMID- 15355875 TI - Characterization of a murine alpha 2 macroglobulin gene expressed in reproductive and cardiovascular tissue. AB - Full-length cDNA for a mouse gene A2-macroglobulin induced by pregnancy (A2mp) was cloned from mesometrial decidua at Gestation Day 10. The 4622-base pair cDNA encodes a protein of 1473 AA with >70% sequence identity and all typical domains of other A2M-family members in humans and rodents, despite unique absence of hepatic expression. The bait region is most distinct and has the greatest sequence similarity with rat acute-phase A2m. Northern blotting, reverse transcription and real-time-PCR, and in situ hybridization studies using C57Bl/6 mice revealed uterine induction of A2mp during decidualization and strong, midgestational association with modifying spiral arteries. Ovaries, testes, lactating mammary glands, heart, and kidney were the only additional organs with A2mp expression that was localized to granulosa and cumulus cells in secondary follicles; primary seminiferous epithelium, including Sertoli cells, mammary alveolar, and ductal epithelium; cardiac endothelium; and renal collecting tubules, respectively. Infusion of native human A2M into pregnant alymphoid or interferon-gamma gene-ablated mice overcame blocks to pregnancy-induced spiral artery modification in these strains. Activated human A2M was also effective, suggesting mechanisms independent of proteinase inhibition. Identification of cytokines, growth factors, or other molecules bound to A2MP should provide new insights into decidualization, spiral artery modification, and cardiovascular adaptation to pregnancy. PMID- 15355876 TI - Fetal programming: testosterone exposure of the female sheep during midgestation disrupts the dynamics of its adult gonadotropin secretion during the periovulatory period. AB - Prenatal exposure of the female sheep to excess testosterone (T) leads to hypergonadotropism, multifollicular ovaries, and progressive loss of reproductive cycles. We have determined that prenatal T treatment delays the latency of the estradiol (E2)-induced LH surge. To extend this finding into a natural physiological context, the present study was conducted to determine if the malprogrammed surge mechanism alters the reproductive cycle. Specifically, we wished to determine if prenatal T treatment 1) delays the onset of the preovulatory gonadotropin surge during the natural follicular phase rise in E2, 2) alters pulsatile LH secretion and the dynamics of the secondary FSH surge, and 3) compromises the ensuing luteal function. Females prenatally T-treated from Day 60 to Day 90 of gestation (147 days is term) and control females were studied when they were approximately 2.5 yr of age. Reproductive cycles of control and prenatally T-treated females were synchronized with PGF2alpha, and peripheral blood samples were collected every 2 h for 120 h to characterize cyclic changes in E2, LH, and FSH and then daily for 14 days to monitor changes in luteal progesterone. To assess LH pulse patterns, blood samples were also collected frequently (each 5 min for 6 h) during the follicular and luteal phases of the cycle. The results revealed that, in prenatally T-treated females, 1) the preovulatory increase in E2 was normal; 2) the latencies between the preovulatory increase in E2 and the peaks of the primary LH and FSH surges were longer, but the magnitudes similar; 3) follicular-phase LH pulse frequency was increased; 4) the interval between the primary and secondary FSH surges was reduced but there was a tendency for an increase in duration of the secondary FSH surge; but 5) luteal progesterone patterns were in general unaltered. Thus, exposure of the female to excess T before birth produces perturbances and maltiming in periovulatory gonadotropin secretory dynamics, but these do not produce apparent defects in cycle regularity or luteal function. To reveal the pathologies that lead to the eventual subfertility arising from excess T exposure during midgestation, studies at older ages must be conducted to assess if there is progressive disruption of neuroendocrine and ovarian function. PMID- 15355877 TI - 17alpha-ethinylestradiol reduces the competitive reproductive fitness of the male guppy (Poecilia reticulata). AB - Whether endocrine disruption in an individual male is actually translated into reduced reproductive success in a natural competitive environment is extremely difficult to predict. Here, we have used paternity analysis to provide new information on the ability of an endocrine disruptor to deleteriously affect male guppy reproductive fitness by including the effect of intermale competition. Groups of male guppies were exposed to 10.5, 44.4, or 112 ng/L of the synthetic estrogen 17alpha-ethinylestradiol (EE2) from birth to adulthood. Subsequently, an exposed male competed against an unexposed male for the opportunity to fertilize a receptive female. The successful males siring the majority of the offspring in each brood were then identified using microsatellites in genetic paternity analysis. Only the highest dose of EE2 produced harmful effects with a significantly female-biased sex ratio, significant reductions in male sperm count, testis weight, body coloration and courtship behavior, and a significant increase in body size. These feminizing effects were translated into a highly significant reduction in fertility, where only 1 of the 17 exposed males sired offspring in competition with unexposed males. The evidence suggests that EE2 treated males have reduced reproductive fitness compared with untreated males, possibly the result of EE2 effects on multiple fitness traits. To our knowledge, this is the first study providing evidence of endocrine disruption at the population level that has included the ecologically highly relevant effect of sexual competition on male reproductive fitness. PMID- 15355878 TI - Clonal organization of proliferating spermatogonial stem cells in adult males of two species of non-human primates, Macaca mulatta and Callithrix jacchus. AB - The present study examines the existence of clonogenic patterns in the proliferation and differentiation of spermatogonial stem cells in two species of non-human primates, the marmoset and the rhesus monkey. We developed a novel approach to detect proliferating spermatogonial clones in whole mounts of seminiferous tubules. Dual fluorescence labeling of bromodeoxyuridine and acrosin in conjunction with confocal microscopy allows the description of the clonogenic and spatial arrangement of proliferating spermatogonia at specific stages of the seminiferous epithelial cycle. Cross-sections of paraffin-embedded tissue were labeled by the same approach. For both monkey species we demonstrate the presence of proliferating spermatogonial clones of variable size at specific stages of the cycle of the seminiferous epithelium. Detailed analysis of the rhesus monkey reveals proliferating Apale spermatogonia at stages VII and IX of the cycle of the seminiferous epithelium, and of proliferating B spermatogonia at stages II, IV, VI, and XII. Proliferating Apale spermatogonia at stages VII and IX of the cycle are organized in pairs or quadruplets. B1 spermatogonia appear as quadruplets or eight-cell clones, and B2 spermatogonia as 8- or 16-cell clones. We conclude that spermatogenesis in the rhesus monkey is initiated by two divisions of duplets or quadruplets of Apale spermatogonia: a first division at stage VII, after which the clones of Apale spermatogonia separate, and a second division at stage IX, which leads to clones of B1 spermatogonia as well as pairs and quadruplets of Apale spermatogonia replenishing the seminiferous epithelium to maintain the original size of the A spermatogonial population. PMID- 15355879 TI - Fowl (Gallus domesticus) sperm motility depends upon mitochondrial calcium cycling driven by extracellular sodium. AB - A relationship between extracellular Ca(+2), fowl sperm phospholipase A2 activity, long-chain acylcarnitine content, and motility was demonstrated in previous work. Sperm motility appeared to depend upon Na+-dependent Ca(+2) cycling when sperm were incubated at body temperature without glucose. In the present work, motility decreased as a function of time when sperm were incubated in 2 mM Ca(+2) prepared with either buffered isotonic sucrose or LiCl. However, this effect was less pronounced in the case of LiCl. The sparing effect of Li+ was attributed to the mitochondrial Na+/Ca(+2) exchanger. Motile concentration decreased exponentially in response to micromolar concentrations of CGP 37157, a specific inhibitor of the mitochondrial Na+/Ca(+2) exchanger. KB-R7943 mesylate, an inhibitor of the reverse mode of the Na+/Ca(+2) exchanger, prevented re initiation of motility when exogenous Ca(+2) was added to sperm rendered immotile by incubation with 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, a high-affinity Ca(+2) chelator. The presence of voltage-gated Ca(+2) channels was confirmed by the effect of nifedipine on motile concentration. Neither motile concentration nor straight line velocity was affected by either ouabain or orthovanadate, which inhibit Na+-K+ ATPase and Ca(+2)-ATPase, respectively. In summary, we infer that 1) fowl sperm motility is dependent upon extracellular Ca(+2) cycling through mitochondria; 2) such cycling is dependent upon extracellular Na+; and 3) fowl sperm conserve ATP by moving neither Na+ nor Ca(+2) by active transport. Understanding the relationship between mitochondrial Ca(+2) cycling and ATP production may be applicable to long-term semen storage. PMID- 15355880 TI - Comparison of germ cell mutagenicity in male CYP2E1-null and wild-type mice treated with acrylamide: evidence supporting a glycidamide-mediated effect. AB - Acrylamide is an animal carcinogen and probable human carcinogen present in appreciable amounts in heated carbohydrate-rich foodstuffs. It is also a germ cell mutagen, inducing dominant lethal mutations and heritable chromosomal translocations in postmeiotic sperm of treated mice. Acrylamide's affinity for male germ cells has sometimes been overlooked in assessing its toxicity and defining human health risks. Previous investigations of acrylamide's germ cell activity in mice showed stronger effects after repeated administration of low doses compared with a single high dose, suggesting the possible involvement of a stable metabolite. A key oxidative metabolite of acrylamide is the epoxide glycidamide, generated by cytochrome P4502E1 (CYP2E1). To explore the role of CYP2E1 metabolism in the germ cell mutagenicity of acrylamide, CYP2E1-null and wild-type male mice were treated by intraperitoneal injection with 0, 12.5, 25, or 50 mg acrylamide (5 ml saline)(-1) kg(-1) day(-1) for 5 consecutive days. At defined times after exposure, males were mated to untreated B6C3F1 females. Females were killed in late gestation and uterine contents were examined. Dose related increases in resorption moles (chromosomally aberrant embryos) and decreases in the numbers of pregnant females and the proportion of living fetuses were seen in females mated to acrylamide-treated wild-type mice. No changes in any fertility parameters were seen in females mated to acrylamide-treated CYP2E1 null mice. Our results constitute the first unequivocal demonstration that acrylamide-induced germ cell mutations in male mice require CYP2E1-mediated epoxidation of acrylamide. Thus, CYP2E1 polymorphisms in human populations, resulting in variable enzyme metabolic activities, may produce differential susceptibilities to acrylamide toxicities. PMID- 15355881 TI - Reduced collagen and ascorbic acid concentrations and increased proteolytic susceptibility with prelabor fetal membrane rupture in women. AB - Prelabor rupture of the fetal membranes affects approximately 10% of women at term, resulting in an increased risk of maternal and neonatal infection. Evidence suggests that membrane rupture is related to biochemical processes involving the extracellular matrix of the membranes. We tested the hypothesis that prelabor ruptured membranes are characterized by reduced collagen concentrations, altered collagen cross-link profiles, and increased concentrations of biomarkers of oxidative damage. We also set out to determine whether these effects are modulated by ascorbic acid status. In a case-control study, we explored the role that ascorbic acid, oxidative stress, collagen, and collagen cross-links play in determining membrane integrity and developed a functional assay to assess membrane proteolytic susceptibility. Prelabor ruptured membrane had a reduced ascorbic acid concentration in comparison with controls while protein carbonyl and malondialdehyde concentrations were increased. Collagen concentrations were also reduced in prelabor ruptured membrane, and while the concentration of collagen cross-links was not significantly different between prelabor and timely ruptured membrane, there was a regional variation in cross-link ratio within the amniotic sac. Proteolytic resistance in vitro was reduced in prelabor ruptured membrane and also exhibited regional variation within the amniotic sac. Our findings are strongly supportive of a role for the enhanced degradation of membrane collagen in the determination of prelabor rupture of fetal membranes. The formation of the rupture initiation site is a function of a regional variation in collagen cross-link ratio. Tissue ascorbic acid status may be an important mediator of these processes. PMID- 15355882 TI - Contribution of phospholipase D in endothelin-1-mediated extracellular signal regulated kinase activation and proliferation in rat uterine leiomyoma cells. AB - Endothelin (ET)-1 is a mitogenic factor in numerous cell types, including rat myometrial cells. In the present study, we investigated the potential role of ET 1 in the proliferation of tumoral uterine smooth muscle cells (ELT-3 cells). We found that ET-1 exerted a more potent mitogenic effect in ELT-3 cells than in normal myometrial cells, as indicated by the increase in [3H]thymidine incorporation, cell number, and bromodeoxyuridine incorporation. The ET-1 was more efficient than platelet-derived growth factor and epidermal growth factor to stimulate proliferation. The ET-1-mediated cell proliferation was inhibited in the presence of U0126, a specific inhibitor of (mitogen-activated protein kinase ERK kinase), indicating that extracellular signal-regulated kinase (ERK) activation is involved. Additionally, ET-1 induced the activation of phospholipase (PL) D, leading to the synthesis of phosphatidic acid (PA). The ET 1-induced activation of PLD was twofold higher in ELT-3 cells compared to that in normal cells. The two cell types expressed mRNA for PLD1a and PLD2, whereas PLD1b was expressed only in ELT-3 cells. The exposure of cells to butan-1-ol reduced ET 1-mediated production of PA by PLD and partially inhibited ERK activation and DNA synthesis. Addition of exogenous PLD or PA in the medium reproduced the effect of ET-1 on ERK activation and cell proliferation. Collectively, these data indicate that ET-1 is a potent mitogenic factor in ELT-3 cells via a signaling pathway involving a PLD-dependent activation of ERK. This highlights the potential role of ET-1 in the development of uterine leiomyoma, and it reinforces the role of PLD in tumor growth. PMID- 15355883 TI - Clinical trial registration: a statement from the International Committee of Medical Journal Editors. PMID- 15355884 TI - Fluorinated 2-(4-amino-3-methylphenyl)benzothiazoles induce CYP1A1 expression, become metabolized, and bind to macromolecules in sensitive human cancer cells. AB - Fluorinated 2-(4-amino-3-methylphenyl)benzothiazoles possess potent antiproliferative activity against certain cancer cells, similar to the unfluorinated 2-(4-amino-3-methylphenyl)benzothiazole (DF 203, NSC 674495). In "sensitive" cancer cells, DF 203 is metabolized by, can induce expression of, and binds covalently to CYP1A1. Metabolism appears to be essential for its antiproliferative activity through DNA adduct formation. However, a biphasic dose response relationship compromises its straightforward development as a chemotherapeutic agent. We investigated whether fluorinated benzothiazoles inhibit cancer cell growth without the biphasic dose-response, and whether the fluorinated benzothiazoles are also metabolized into reactive species, with binding to macromolecules in sensitive cancer cells. One fluorinated benzothiazole, 2-(4-amino-methylphenyl)-5-fluorobenzothiazole (5F 203, NSC 703786) did exhibit potent, antiproliferative activity without a biphasic dose response. The fluorinated benzothiazoles were also metabolized only in cells, which subsequently showed evidence of cell death. We used microsomes from genetically engineered human B-lymphoblastoid cells expressing cytochromes P450 (CYP1A1, CYP1A2, or CYP1B1) to clarify the basis for fluorinated benzothiazole metabolism. 5F 203 induced CYP1A1 and CYP1B1 mRNA expression in sensitive breast and renal cancer cells, whereas 5F 203 induced CYP1A1 mRNA but not CYP1B1 mRNA expression in sensitive ovarian cancer cells. 5F 203 did not induce CYP1A1 or CYP1B1 mRNA expression in any "resistant" cancer cells. The fluorinated benzothiazoles induced CYP1A1 protein expression exclusively in sensitive cells. [14C]5F 203 bound substantially to subcellular fractions in sensitive cells but only minimally in resistant cells. These data are concordant with the antiproliferative activity of fluorinated benzothiazoles deriving from their ability to become metabolized and bind to macromolecules within sensitive cells. PMID- 15355885 TI - Differences in FMO2*1 allelic frequency between Hispanics of Puerto Rican and Mexican descent. AB - A polymorphism for the phase I drug-metabolizing enzyme, flavin-containing monooxygenase isoform 2 (FMO2), encoding either truncated inactive protein, FMO2X472 (FMO2.2A), or full-length active enzyme, FMO2Q472 (FMO2.1), is known and exhibits significant interethnic differences in allelic frequency. FMO2 is the major or sole FMO isoform expressed in the lung of most mammals, including nonhuman primates. To date, FMO2.1 has been found only in African-American and Hispanic populations, rendering individuals with this allele subject to drug metabolism that is potentially different from that of the general population. Approximately 26% of African-Americans (n = 180) possess the FMO2*1 allele. In preliminary studies, we initially estimated that 5% of Hispanics (n = 40) have the FMO2*1 allele, but access to large cohorts of individuals of defined national origin has allowed us to determine the occurrence among Mexican-American and Puerto Rican-American groups. We used allele-specific genotyping to detect FMO2*1 from 632 Hispanic individuals, including 280 individuals of Mexican origin and 327 individuals of Puerto Rican origin. Statistical analysis indicated that results from Mexican (five sample sources) and Puerto Rican (three sample sources) samples were consistent with the hypothesis of homogeneity within each group from different sources. Data were subsequently pooled across sources to test for evidence of a difference in occurrence of FMO2*1 between ethnic groups. There was strong evidence (p = 0.0066) that FMO2*1 is more common among Puerto Ricans (7%) than among individuals of Mexican descent (2%). The overall occurrence of FMO2*1 among Hispanics of all origins is estimated to be between 2 and 7%. PMID- 15355886 TI - Detection characteristics in randomized prostate screening. PMID- 15355887 TI - Treatment of relapsed central nervous system lymphoma with high-dose methotrexate. AB - PURPOSE: Over the past decade, high-dose methotrexate has emerged as the single most effective agent in the initial treatment of primary nervous system lymphoma. However, the majority of patients who respond initially to treatment relapse. The optimal management of these patients has not been determined. We performed a multicenter, retrospective study of high-dose methotrexate in patients with relapsed central nervous system lymphoma. EXPERIMENTAL DESIGN: Patients with relapsed disease were eligible if they achieved a complete response to initial treatment with methotrexate-based chemotherapy or received methotrexate after gross total resection or interstitial radiation. All of the patients were retreated with a regimen containing high-dose methotrexate (>/=3 g/m(2)). RESULTS: Twenty-two patients with a median age of 58 years were included in the study. Overall response rates were 91% to first salvage (20 of 22 patients) and 100% to second salvage (4 of 4 patients). Median survival was 61.9 months after first relapse (95% confidence interval, 42.1- infinity ) and 91.9 months overall (95% confidence interval, 47.2- infinity ). Toxicity was primarily hematologic with 10 episodes of grade 3 or 4 toxicity during 566 cycles of chemotherapy. CONCLUSIONS: These results indicate that high-dose methotrexate remains effective for relapsed central nervous system lymphoma in patients who initially respond to methotrexate and raise the possibility of deferring more toxic salvage regimens in this select group of patients. PMID- 15355888 TI - Mouse mammary tumor virus-like viral infection and human breast cancer. PMID- 15355889 TI - Pilot study of the mechanism of action of preoperative trastuzumab in patients with primary operable breast tumors overexpressing HER2. AB - PURPOSE: To elucidate the mechanism by which trastuzumab, a humanized monoclonal antibody against HER2 with proven survival benefit in women with HER2-positive metastatic breast cancer, mediates its antitumor activity. EXPERIMENTAL DESIGN: A pilot study including 11 patients with HER2-positive tumors treated in a neo adjuvant setting with trastuzumab was performed. Trastuzumab was administered i.v. at a dose of 4 mg/kg followed by three weekly i.v. doses of 2 mg/kg. The primary tumor was surgically removed 7 days after the last treatment. Surgical samples, tumor biopsies, and lymphocytes from these patients were collected for biological studies. RESULT: Clinical data indicated one complete pathological remission and four partial remissions using RECIST (Response Evaluation Criteria in Solid Tumors). Trastuzumab was well tolerated and neither serious adverse events nor changes in cardiac function were observed during this short-term treatment and after surgery. The biological data showed that, independent of response, (a) all patients showed high levels of circulating trastuzumab; (b) saturating level of trastuzumab was present in all of the tumors; (c) no down modulation of HER2 was observed in any tumors; (d) no changes in vessel diameter was observed in any tumors; (e) no changes in proliferation was observed in any tumors; and (f) a strong infiltration by lymphoid cells was observed in all cases. Patients with complete remission or partial remission were found to have a higher in situ infiltration of leukocytes and a higher capability to mediate in vitro antibody-dependent cellular cytotoxicity activity. CONCLUSIONS: The results of this pilot study argue against trastuzumab activity in patients through down modulation of HER2 but in favor of antibody-dependent cellular cytotoxicity guiding efforts to optimize the use of trastuzumab in breast cancer patients. PMID- 15355890 TI - Clonal isolation of different strains of mouse mammary tumor virus-like DNA sequences from both the breast tumors and non-Hodgkin's lymphomas of individual patients diagnosed with both malignancies. AB - PURPOSE: In a previous study, we had detected the presence of mouse mammary tumor virus (MMTV)-like envelope (ENV) gene sequences in both the breast tumors and non Hodgkin's lymphoma tissue of two of our breast tumor patients who had been diagnosed simultaneously with both malignancies. The aim of this study was to determine if MMTV-like DNA sequences are present in the breast tumors and non Hodgkin's lymphomas of additional patients suffering from both malignancies and if so to characterize these sequences in detail. EXPERIMENTAL DESIGN: DNA was extracted from formalin-fixed, paraffin-embedded tissue sample blocks of breast tumors and non-Hodgkin's lymphomas from patients suffering from both malignancies. A 250-bp region of the MMTV ENV gene and a 630-bp region of the MMTV long terminal repeat (LTR) open reading frame (ORF) that encodes the MMTV superantigen (sag) gene were amplified by PCR from the isolated DNA. Amplified products were analyzed by Southern blotting, cloned, and sequenced. RESULTS: MMTV like ENV and LTR sequences were detected in both the breast tumors and non Hodgkin's lymphomas of 6 of 12 patients suffering from both malignancies. A novel mutant of the MMTV ENV gene was identified in these patients. Characterization of the MMTV-like LTR highly variable sag sequences revealed total or nearly total identity to three distinct MMTV proviruses from two different branches of the MMTV phylogenetic tree. CONCLUSIONS: The presence of MMTV-like ENV and LTR sequences in both the breast tumors and non-Hodgkin's lymphomas of 6 additional patients suggests a possible involvement of these sequences in these two malignancies. MMTV-like LTR sequence homology to different MMTV proviruses revealed the presence of more than one strain of MMTV-like sequences in each individual suggesting the possibility of multiple infections in these patients. PMID- 15355891 TI - Nuclear localization of apoptosis protease activating factor-1 predicts survival after tumor resection in early-stage non-small cell lung cancer. AB - The proapoptotic protein apoptosis protein activating factor-1 (Apaf-1), which is normally located in the cytoplasm, can translocate to the nucleus before non small cell lung carcinoma (NSCLC) cells manifest signs of apoptosis such as mitochondrial damage, caspase activation, or chromatin condensation. This may indicate a stage of imminent apoptosis. Importantly, we found that 24% (15 of 62) of resected stage I NSCLC (T(1)N(0)M(0) or T(2)N(0)M(0)), manifested a marked nuclear localization of Apaf-1 (Apaf-1(Nuc)), as compared with the mostly cytoplasmic localization of Apaf-1 found in the remaining tumors (Apaf-1(Cyt)). After a median follow-up of 6.31 years, the actuarial 5-year overall survival rates were 89% (56-98%) in the Apaf-1(Nuc) group and 54% (36-71%) in the Apaf 1(Cyt) group (P = 0.039). No correlation between the subcellular localization of Apaf-1 and that of p53 and Hsp70 could be established. Thus, the subcellular location of Apaf-1 (but not that of p53 or Hsp70) constitutes an accurate prognostic factor for overall survival in NSCLC. PMID- 15355892 TI - HER-2 amplification, HER-1 expression, and tamoxifen response in estrogen receptor-positive metastatic breast cancer: a southwest oncology group study. AB - PURPOSE: Preclinical data indicate that expression of the ErbB family of receptors, such as HER-2 and HER-1 (EGFR) may be involved in endocrine resistance. Evidence of resistance from clinical studies has been inconsistent. The present study examined whether HER-2 gene amplification or HER-1 expression predicted response to tamoxifen. PATIENTS AND METHODS: Three hundred and forty nine patients had estrogen receptor (ER)-positive breast cancer and received daily tamoxifen as initial therapy for advanced disease. HER-2 gene amplification, detected by fluorescence in situ hybridization, and HER-1 expression, evaluated by immunohistochemistry, was determined on 136 and 204 patients, respectively. RESULTS: HER-2 amplification was correlated with lower ER (P = 0.02), HER-1 positivity (P = 0.004), and HER-2 protein overexpression (P < 0.00001). The response rate was 56% for HER-2 non-amplified versus 47% for HER-2 amplified tumors (P = 0.38), and 58% for HER-1-negative versus 36% for HER-1 positive (P = 0.05). Time to treatment failure (TTF) was 7 months for non amplified HER-2 tumors and 5 months (P = 0.007) for amplified HER-2 tumors, and there was a trend toward a better overall survival (OS) in patients with non amplified HER-2 tumors (median 31 versus 25 months, respectively, P = 0.07). For positive versus negative HER-1 tumors, TTF was 4 versus 8 months (P = 0.08) and median survival was 24 versus 31 months (P = 0.41). Combining HER-1 expression and HER-2 gene status, patients with both negative HER-1 expression and non amplified HER-2 had longer TTF (P = 0.001) and OS (P = 0.03) than if either were positive. In multivariate analysis, HER-2 was not an independent factor for TTF and OS, although HER-1 was significant for TTF only (P 18 years old. Stem cell factor and MK concentrations above a defined threshold in serum of NF1 patients are of diagnostic benefit for 96% of patients in the cohort tested. Furthermore, serum from NF1 patients enhanced proliferation of human neurofibroma-derived primary Schwann cells and endothelial cells substantially better than normal serum. CONCLUSIONS: Enhanced circulating growth factor levels contribute to diffuse tumorigenesis in NF1 and may provide the basis for molecular diagnosis. PMID- 15355894 TI - Prognostic significance of p16 protein levels in oropharyngeal squamous cell cancer. AB - PURPOSE: Functional inactivation of p16 is an early and frequent event in head and neck squamous cell cancers. In this study, we sought to determine whether p16 expression is of prognostic importance in oropharyngeal squamous cell carcinoma. EXPERIMENTAL DESIGN: p16 protein expression was evaluated by immunohistochemistry in a tissue microarray composed of 123 oropharyngeal squamous cell cancers with a mean patient follow-up time of 33 months. RESULTS: p16 overexpression was associated with more advanced Tumor-Node-Metastasis stage and higher histologic grade. Despite this association with unfavorable features, p16 overexpression was associated with decreased 5-year local recurrence rates (11 versus 53%) and increased 5-year disease-free survival (62 versus 19%) and overall survival (60 versus 21%). In multivariate analysis, p16 expression status remained an independent prognostic factor for local recurrence, disease-free survival, and overall survival. CONCLUSIONS: In patients with oropharyngeal squamous cell carcinoma, overexpression of p16 as determined by immunohistochemistry is associated with significantly improved prognosis and lower local recurrence rates. PMID- 15355895 TI - A global expression-based analysis of the consequences of the t(4;14) translocation in myeloma. AB - PURPOSE: Our purpose in this report was to define genes and pathways dysregulated as a consequence of the t(4;14) in myeloma, and to gain insight into the downstream functional effects that may explain the different prognosis of this subgroup. EXPERIMENTAL DESIGN: Fibroblast growth factor receptor 3 (FGFR3) overexpression, the presence of immunoglobulin heavy chain-multiple myeloma SET domain (IgH-MMSET) fusion products and the identification of t(4;14) breakpoints were determined in a series of myeloma cases. Differentially expressed genes were identified between cases with (n = 5) and without (n = 24) a t(4;14) by using global gene expression analysis. RESULTS: Cases with a t(4;14) have a distinct expression pattern compared with other cases of myeloma. A total of 127 genes were identified as being differentially expressed including MMSET and cyclin D2, which have been previously reported as being associated with this translocation. Other important functional classes of genes include cell signaling, apoptosis and related genes, oncogenes, chromatin structure, and DNA repair genes. Interestingly, 25% of myeloma cases lacking evidence of this translocation had up regulation of the MMSET transcript to the same level as cases with a translocation. CONCLUSIONS: t(4;14) cases form a distinct subgroup of myeloma cases with a unique gene signature that may account for their poor prognosis. A number of non-t(4;14) cases also express MMSET consistent with this gene playing a role in myeloma pathogenesis. PMID- 15355896 TI - N-cadherin is regulated by activin A and associated with tumor aggressiveness in esophageal carcinoma. AB - PURPOSE: Activin A is a member of the transforming growth factor beta superfamily and plays an important role in the differentiation of embryonic stem cells. We have reported previously that the expression of activin A is associated with lymph node metastasis in esophageal cancer, and our purpose in the current work is to clarify the molecular mechanism of the aggressive behavior of tumors that have high activin A expression. EXPERIMENTAL DESIGN: We have compared the gene expression profiles of human esophageal carcinoma cell lines that were stably transfected with activin beta A, which is a subunit of activin A, with those of control human esophageal carcinoma cell lines, using a cDNA microarray. RESULTS: We found that the expression level of neuronal cadherin (N-cadherin) was higher in the transfectants than in the control cells. N-cadherin was located on the cell surface of the transfectants, irrespective of the expression of epithelial cadherin (E-cadherin), and the expression of N-cadherin mRNA was significantly associated with that of activin beta A mRNA in clinical samples of esophageal carcinoma (n = 51; r = 0.855). A clinicopathologic analysis suggested that expression of N-cadherin mRNA was associated with the depth of tumor wall invasion, and a group of patients with high expression of N-cadherin mRNA showed a significantly poorer prognosis than a group of patients with low N-cadherin expression (P = 0.046). CONCLUSIONS: These results indicate that activin A might mediate the expression of N-cadherin and that this may be associated with depth of invasion and poor prognosis. PMID- 15355897 TI - Melanoma-associated antigens in esophageal adenocarcinoma: identification of novel MAGE-A10 splice variants. AB - PURPOSE: The melanoma-associated antigens (MAGEs) are tumor-specific antigens recognized by cytotoxic T lymphocytes. In this study, expression of MAGE family A members was evaluated during the development of esophageal adenocarcinoma (EA) as potential targets for immunotherapy. EXPERIMENTAL DESIGN: MAGE-A mRNA expression was evaluated in 46 samples including Barrett's metaplasia (BM), dysplasia, and EA using oligonucleotide microarrays. Expression of MAGE-A proteins was confirmed by immunohistochemistry on tissue microarrays containing 59 EA, 11 dysplasia, and 9 BM samples and by Western blot. To further evaluate MAGE-A10 expression, reverse transcription-polymerase chain reaction (RT-PCR) products were sequenced, and protein expression was determined using a specific antibody. RESULTS: Overexpression of MAGE-A1, MAGE-A2b, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A9, MAGE A10, and MAGE-A12 was found in EAs relative to BM on oligonucleotide microarrays. MAGE-A3 overexpression was confirmed by real-time RT-PCR in 21.4% (6 of 28) of esophageal tumors. Immunohistochemistry on tissue microarray revealed MAGE-A proteins in 20.3% (12 of 59) of EAs and MAGE-A10 staining in 16.9% (10 of 59) of EAs. MAGE-A expression was confirmed by Western blot in several esophageal tumors and in two EA cell lines, Flo-1 and Seg-1, whereas Flo-1 also expressed MAGE-A10. Tumors produced from these cell lines in nude mice retained MAGE-A expression. Interestingly, RT-PCR in primary tumors expressing MAGE-A10 protein revealed additional PCR products that were identified as novel MAGE-A10 alternative splice variants using DNA sequencing. CONCLUSIONS: This is the first report of these MAGE-A10 alternative splice sequences, and characterization of MAGE-A expression may provide potential targets for immunotherapy in patients with EA. PMID- 15355898 TI - Effects of the steroidal aromatase inhibitor exemestane and the nonsteroidal aromatase inhibitor letrozole on bone and lipid metabolism in ovariectomized rats. AB - PURPOSE: Exemestane (EXE) and letrozole (LET) are third-generation aromatase inhibitors currently prescribed for postmenopausal hormone-dependent breast cancer. The impact on end organs of estrogen depletion in menopausal women is of significant clinical importance. We studied the effects of EXE, its principal metabolite, 17-hydroexemestane (17-H-EXE), and LET on bone and lipid metabolism in ovariectomized (OVX) rats. EXPERIMENTAL DESIGN: OVX rats were treated by weekly intramuscular injection for 16 weeks with 20, 50, and 100 mg/kg EXE, 20 mg/kg 17-H-EXE, and daily oral gavage of 1 mg/kg LET. At the end of the treatment period, bone mineral density (BMD), the bone resorption marker serum pyridinoline, the bone formation marker serum osteocalcin, bone mechanical properties, histomorphometry, and serum lipid concentrations were determined. RESULTS: Lumbar vertebral and femoral BMD, bending strength of the femur, compressive strength of the fifth lumbar vertebra, and trabecular bone volume were significantly higher in OVX animals given EXE and 17-H-EXE than in OVX controls. EXE and 17-H-EXE significantly reduced an ovariectomy-induced increase in serum pyridinoline and serum osteocalcin. EXE and 17-H-EXE given to OVX rats caused significant reductions of serum cholesterol and low-density lipoprotein cholesterol. In contrast, OVX rats treated with LET had BMD, bone biomarkers, mechanical failure properties, and lipid levels similar to those of OVX controls. CONCLUSIONS: EXE and 17-H-EXE significantly prevent bone loss, enhance bone mechanical strength, and lower serum cholesterol and low-density lipoprotein levels in OVX rats. These protective effects on end-organ function are not seen with the nonsteroidal inhibitor LET. PMID- 15355899 TI - Ionizing radiation causes a dose-dependent release of transforming growth factor alpha in vitro from irradiated xenografts and during palliative treatment of hormone-refractory prostate carcinoma. AB - PURPOSE: Characterize the radiation response for transforming growth factor (TGF) alpha shedding in vitro and in vivo. We also report the shedding of TGF alpha by patients undergoing irradiation for hormone-refractory prostate cancer. EXPERIMENTAL DESIGN: TGF alpha levels were determined by ELISA. DU145 xenografts were established on the flanks of athymic nu/nu mice. Expression of phospho extracellular signal-regulated kinase (ERK)1/2 and phospho-epidermal growth factor receptor (EGFR) and the DNA repair proteins XRCC1 and ERCC1 were determined by Western analyses. RESULTS: Exposure to ionizing radiation results in a dose-dependent release of TGF alpha. Once released, TGF alpha stimulates EGFR-ERK1/2 signaling in unirradiated cells. Blockade of the EGFR with the tyrphostin AG1478 eliminates the up-regulation XRCC1 and ERCC1 by TGF alpha or irradiation. After irradiation, cells are refractory to additional transactivation of EGFR by additional irradiation for 8 to 12 hours. Irradiation during this refractory period does not increase the expression of XRCC1 or ERCC1. Ligand activation of EGFR is maintained during the refractory period. Irradiation of DU145 xenografts also results in the activation of ERK1/2, release of TGF alpha, and a similar refractory period. Ionizing irradiation also results in the release of TGF alpha for patients undergoing radiation therapy for hormone refractory prostate cancer. CONCLUSIONS: Irradiation results in a dose-dependent increase in TGF alpha capable of enhancing the growth of DU145 xenografts. TGF alpha is also shed following radiation therapy of patients treated for hormone refractory prostate cancer. Radiation transactivation of the EGFR produces a radio-refractory period, which lasts for several hours. During this period, additional irradiation fails to induce XRCC1, ERCC1, or additional TGF alpha release. PMID- 15355900 TI - Phase II study of the antiangiogenic agent SU5416 in patients with advanced soft tissue sarcomas. AB - PURPOSE: SU5416 (semaxanib) is a small molecule inhibitor of the vascular endothelial growth factor (VEGF) receptor-2 and KIT receptor tyrosine kinases. This Phase II study was conducted to investigate the safety and efficacy of SU5416 for patients with soft tissue sarcomas. EXPERIMENTAL DESIGN: Thirteen patients with locally advanced or metastatic soft tissue sarcomas were treated with SU5416 via intravenous infusion at a dose of 145 mg/m(2) twice weekly. In selected cases tumor biopsies were taken before and after 2 months of treatment. RESULTS: The median progression-free survival was 1.8 months. Median overall survival was 22.8 months. No objective tumor responses were observed. There was evidence of shorter survival among patients with high baseline urine VEGF levels (P = 0.04). No grade 4 toxicities were observed. The most common grade 3 toxicities were headache and thrombosis. Other less serious toxicities included fatigue, nausea, and abdominal pain. The median systolic blood pressure increased from 118 mmHg at baseline to 133 after 1 month of treatment (P = 0.01). Post treatment tumor biopsies showed no significant decreases in VEGF receptor phosphorylation compared with baseline in 3 evaluable patients. One patient with gastrointestinal stromal tumor who had rapid progression during SU5416 treatment was subsequently treated with another KIT inhibitor, imatinib mesylate, and had a partial response lasting >36 months. CONCLUSIONS: SU5416 was relatively well tolerated but did not demonstrate significant antitumor activity against advanced soft tissue sarcoma. Correlative studies suggest that VEGF receptor or KIT inhibition was incomplete in at least some cases, providing a possible explanation for the observed lack of activity. PMID- 15355901 TI - Carbon ion radiotherapy for unresectable sacral chordomas. AB - PURPOSE: The purpose is to evaluate the efficacy and toxicity of carbon ion radiotherapy for unresectable sacral chordomas. EXPERIMENTAL DESIGN: We performed a retrospective analysis of 30 patients with unresectable sacral chordomas treated with carbon ion radiotherapy at the Heavy Ion Medical Accelerator in Chiba, Japan. Twenty-three patients presented with no prior treatment, and the remaining 7 patients had locally recurrent disease following previous surgical resection. The median clinical target volume was 546 cm(3). The applied carbon ion dose ranged from 52.8 to 73.6 GyE (gray equivalent, median 70.4) in 16 fixed fractions over 4 weeks. RESULTS: At median follow-up of 30 months (range, 9 to 87 months), 26 patients were still alive and 24 patients remained continuously disease-free. Overall and cause-specific survival rates at 5 years were 52 and 94%, respectively. The overall local control rate at 5 years was 96%. Two patients experienced severe skin/soft tissue complications requiring skin grafts. No other treatment-related surgical interventions, including colostomy or urinary diversion, were carried out. All patients have remained ambulatory and able to stay at home after carbon ion radiotherapy. CONCLUSIONS: Carbon ion radiotherapy is effective and safe in the management of patients with unresectable sacral chordomas and offers a promising alternative to surgery. PMID- 15355902 TI - A Phase I trial of TNFerade biologic in patients with soft tissue sarcoma in the extremities. AB - PURPOSE: TNFerade is a second-generation replication-deficient adenovector carrying a transgene encoding human tumor necrosis factor alpha under control of a radiation- induced promoter. The objective of this study was to assess the tolerance of combining TNFerade and radiation therapy in patients with soft tissue sarcomas of the extremity. EXPERIMENTAL DESIGN: TNFerade was administered in combination with single-daily fractionated radiation therapy in 14 patients with soft tissue sarcoma of the extremities. Three escalating dose levels of TNFerade (4 x 10(9) -4 x 10(11) particle units) were planned, given in 1 log increments by intratumoral injections, twice weekly during week 1 and once weekly during weeks 2-5 of radiation therapy. RESULTS: TNFerade was well tolerated with no dose-limiting toxicities noted. Grade 1-2 chills (50.0%), fever (43.0%), fatigue (36.0%), and flu-like symptoms (21.0%) were the most common side effects. Serum-tumor necrosis factor alpha levels were low in all of the patients (<15 pg/mL). No patients had virus-detected blood, sputum, or urine cultures. Of the 13 evaluable patients, 11 received TNFerade preoperatively, and 2 received the treatment for palliation. Eleven patients (85%) showed objective or pathological tumor responses (2 complete and 9 partial), and 1 had stable disease. Partial responses were achieved despite some of these tumors being very large (up to 675 cm(2)). Of the 11 patients who underwent surgery, 10 (91%) showed a pathological complete response/partial response. CONCLUSION: TNFerade + radiation therapy was well tolerated in the treatment of patients with soft-tissue sarcoma of the extremity. The high number of objective responses observed warrants additional studies of this approach in a larger controlled prospective trial. PMID- 15355903 TI - Phase II study of feasibility of dose-dense FEC followed by alternating weekly taxanes in high-risk, four or more node-positive breast cancer. AB - PURPOSE: To develop a potentially superior adjuvant chemotherapy regimen, we conducted a pilot study of dose-dense 5-fluorouracil, epirubicin, and cyclophosphamide (FEC) followed by weekly alternating taxanes. The primary objective was to determine the feasibility of the regimen; the secondary objective was to estimate the disease-free and overall survival. EXPERIMENTAL DESIGN: Patients with >/=4 node-positive breast cancer were studied. Treatment consisted of FEC at 500/100/500 mg/m(2), respectively, x6 at two-week intervals with granulocyte colony-stimulating factor, followed by weekly paclitaxel (80 mg/m(2)) alternating with docetaxel (35 mg/m(2)) x18. RESULTS: Between November 2001 and January 2003, 44 patients were enrolled. Median age was 46 years (range, 26-63 years), median number of positive nodes was 9 (range, 4-32), and median tumor size was 2.5 cm (range, 0.6-11.0 cm). Because of unexpected toxicities, the study was stopped when 17 (39%) had fully completed all of the planned treatment. Two of 17 (12%) developed grade 4 pericardial/grade 3 bilateral pleural effusions at treatment completion; both required pericardial window. The remaining patients were treated with taxanes using one of several standard dose and schedule combinations. Furthermore, 4 of 44 (9%) developed pneumonitis attributed to the FEC regimen. Hospital admissions were required for 12 of 44 (27%); 3 of 44 (7%) required blood transfusions. There were no treatment related deaths. Median disease-free and overall survival will not be estimatable because of early closure of study. CONCLUSION: FEC x6 at 2-week intervals followed by 18 weeks of alternating taxanes is not feasible at the doses tested. Other strategies are needed to improve adjuvant systemic chemotherapy. PMID- 15355904 TI - Human telomerase reverse transcriptase gene expression and the surgical management of suspicious thyroid tumors. AB - PURPOSE: Patients with a preoperative cytologic diagnosis of a suspicious thyroid nodule present a therapeutic dilemma because surgery differs for benign and malignant lesions. To address this issue, several molecular markers, including human telomerase reverse transcriptase (TERT), have been tested as markers of thyroid cancer. Because most studies select cases falling into well-defined categories to test new markers, they may overestimate their discriminatory power when applied to samples that are difficult to classify. Fine-needle aspirates (FNAs) of the thyroid with indeterminate cytology are an example of such cases. EXPERIMENTAL DESIGN: We examined whether assessing TERT mRNA by reverse transcription-PCR could have improved the surgical management in a cohort of 100 patients undergoing thyroidectomy for indeterminate FNA results. RESULTS: Ninety percent of 48 cancers were TERT positive, as were 35% of 52 benign lesions. When 10 cases with concomitant lymphocytic thyroiditis were excluded, the overall sensitivity of TERT was 91% (95% confidence interval, 80-98%) and specificity was 79% (64-90%). No clinical or tumor variable contributed to the predictive ability of TERT except for tumor size, which added only marginally. Basing the surgical approach on the TERT assay alone would have reduced lobectomies performed for malignant disease from 11 to 4 cases and reduced total thyroidectomies for benign lesions from to 15 to 9, an overall 50% reduction in suboptimal treatment. CONCLUSIONS: The overall performance of preoperative differential diagnosis for thyroid tumors with indeterminate FNA results can be substantially improved by the inclusion of molecular markers such as TERT. PMID- 15355905 TI - Estrogen receptor beta (ER beta) level but not its ER beta cx variant helps to predict tamoxifen resistance in breast cancer. AB - The antiestrogen tamoxifen, a major endocrine therapy of estrogen receptor (ER) positive breast cancer, is nevertheless inefficient in 30 to 40% of cases for unknown reasons. We retrospectively studied 50 ER-positive primary breast carcinomas. All of the patients had received tamoxifen as the only adjuvant therapy. They were divided into two groups depending on whether they relapsed within 5 years (16 tamoxifen-resistant cases) or did not relapse within 5 years (34 tamoxifen-sensitive cases). The expression of total ER beta protein, and of ER beta cx protein, was estimated anonymously in formalin-fixed, paraffin embedded tumor sections, by using specific antibodies and quantifiying nuclear immunostaining with a computer image analyzer. All of the tumors were found to be HER-2/neu-negative by immunohistochemistry. Univariate analysis showed that Scarff-Bloom-Richardsson grade modified by Elston (SBR grade; P < 0.001), tumor size (P = 0.042), and MIB-1 proliferation index (P = 0.02) were significantly higher in tamoxifen-resistant tumors. A low level of total ER beta, whether in percentage of positive cells or in quantitative immunocytochemical (QIC) score, was also associated with tamoxifen resistance (P = 0.004). ER beta cx expression and lymph node status were similar between the two groups. The expression of ER beta in the total population was positively correlated with ER beta cx (r = 0.63, P < 0.001), and was independent of the other parameters. In a multivariate analysis, ER beta expression was the most important variable (P = 0.001), followed by SBR grade (I+II versus III; P = 0.008), and MIB-1 (P = 0.016). To conclude, tamoxifen resistance is associated with classical variables of aggressive tumors (high SBR grade, proliferation index, and tumor size) but not with node invasiveness. Low ER beta level is an additional independent marker, better than ER alpha level, to predict tamoxifen resistance. PMID- 15355906 TI - Quantification of colorectal cancer micrometastases in lymph nodes by nested and real-time reverse transcriptase-PCR analysis for carcinoembryonic antigen. AB - PURPOSE: Reverse-transcriptase PCR (RT-PCR) assays for carcinoembryonic antigen (CEA) have been described to identify lymph node micrometastases. These assays are not quantitative and can be confounded by false-positive results. The purpose of this study was to determine whether quantification of CEA in lymph nodes could more readily identify clinically relevant groups. EXPERIMENTAL DESIGN: Specimens included 400 lymph nodes from 64 patients undergoing colon resections. Specimens were tested by immunohistochemistry and by RT-PCR using nested primers for CEA. Specimens from 59 patients that were positive by nested RT-PCR were further quantified by detection of CEA mRNA fluorescence increase at a threshold PCR cycle. RESULTS: CEA was detected by nested RT-PCR analysis in 4 of 34 (12%) nodes of nonneoplastic disease, 2 of 13 (15%) nodes from T(1)N(0) patients, 32 of 81 (40%) nodes of T(2)N(0) patients, 49 of 109 (45%) nodes from T(3)N0 patients, and 92 of 163 (56%) nodes from T(1-4)N(1-2) patients. The overall presence of any RT PCR-detectable CEA in nodes did not differentiate patient groups. Immunohistochemistry was positive in nodes from 7% of T(3)N(0) patients and 100% of T(1-3)N(1-2) patients. CEA quantification revealed that 0 of 7 patients with nonneoplastic disease and 2 of 17 (12%) patients with stage I T(1-2)N(0) cancers had one or more lymph nodes with >/=1.0 x 10(2) CEA transcripts per sample. In contrast, 4 of 13 (31%) patients with stage II T(3)N(0) cancer and 10 of 22 (45%) stage III patients with known metastases had lymph nodes with >/=1.0 x 10(2) CEA transcripts. CONCLUSIONS: These data suggest that quantification of CEA levels in lymph nodes may more accurately identify patients at risk for cancer recurrence than does routine nested RT-PCR or immunohistochemistry. PMID- 15355907 TI - Amplification of the BRCA2 pathway gene EMSY in sporadic breast cancer is related to negative outcome. AB - DNA amplification at band q13 of chromosome 11 is common in breast cancer, and CCND1 and EMS1 remain the strongest candidate genes. However, amplification patterns are consistent with the existence of four cores of amplification, suggesting the involvement of additional genes. Here we present evidence strongly suggesting the involvement of the recently characterized EMSY gene in the formation of the telomeric amplicon. EMSY maps at 11q13.5, 100 kb centromeric to the GARP gene, which has been mapped within the core of the distal amplicon. The EMSY protein was shown to interact with BRCA2 and has a role in chromatin remodeling. This makes EMSY a strong candidate oncogene for the 11q13.5 amplicon. DNA amplification was studied in a total of 940 primary breast tumors and 39 breast cancer cell lines. Amplification profiles were consistent with the EMSY GARP locus being amplified independently of CCND1 and/or EMS1. EMSY RNA expression levels were studied along with those of five other genes located at 11q13.5 by real-time quantitative PCR in the 39 cell lines and a subset of 65 tumors. EMSY overexpression correlated strongly with DNA amplification in both primary tumors and cell lines. In a subset of 296 patients, EMSY amplification was found by both uni- and multivariate analyses to correlate with shortened disease-free survival. These data indicate that EMSY is a strong candidate oncogene for the 11q13.5 amplicon. PMID- 15355908 TI - Casein kinase II alpha subunit and C1-inhibitor are independent predictors of outcome in patients with squamous cell carcinoma of the lung. AB - PURPOSE: Gene expression profiling has been shown to be a valuable tool for prognostication and identification of cancer-associated genes in human malignancies. We aimed to identify potential prognostic marker(s) in non-small cell lung cancers using global gene expression profiles. EXPERIMENTAL DESIGN: Twenty-one previously untreated patients with non-small cell lung cancer were analyzed using the Affymetrix GeneChip high-density oligonucleotide array and comparative genomic hybridization. Identified candidate genes were validated in an independent cohort of 45 patients using quantitative real-time reverse transcription-PCR and Western blot analyses. Follow-up data for these patients was collected and used to assess outcome correlations. RESULTS: Hierarchical clustering analysis yielded three distinct subgroups based on gene expression profiling. Cluster I consisted of 4 patients with adenocarcinoma and 1 with squamous cell carcinoma (squamous cell carcinoma); clusters II and III consisted of 6 and 10 patients with squamous cell carcinoma, respectively. Outcome analysis was performed on the cluster groups containing solely squamous cell carcinoma, revealing significant differences in disease-specific survival rates. Moreover, patients having a combination of advanced Tumor-Node-Metastasis stage and assigned to the poor prognosis cluster group (cluster II) had significantly poorer outcomes. Comparative genomic hybridization analysis showed recurrent chromosomal losses at 1p, 3p, 17, 19, and 22 and gains/amplifications at 3q, 5p, and 8q, which did not vary significantly between the cluster groups. We internally and externally validated a subset of 11 cluster II (poor prognosis) specific genes having corresponding chromosomal aberrations identified by comparative genomic hybridization as prognostic markers in an independent cohort of patients with lung squamous cell carcinoma identifying CSNK2A1 and C1-Inh as independent predictors of outcome. CONCLUSION: CSNK2A1 and C1-Inh are independent predictors of survival in lung squamous cell carcinoma patients and may be useful as prognostic markers. PMID- 15355909 TI - Expression of transforming growth factor beta isoforms and their receptors in malignant fibrous histiocytoma of soft tissues. AB - PURPOSE: Transforming growth factor beta (TGF-beta) is a multifunctional growth factor that variably affects proliferation, differentiation, and extracellular matrix formation. Little information is currently available on the TGF-beta expression in malignant fibrous histiocytoma (MFH). The aims of the present study were to investigate the expression of TGF-beta isoforms and their receptors in human MFH specimens. EXPERIMENTAL DESIGN: The expression of TGF isoforms, and TGF beta receptors (TGF-beta R1 and -beta R2) were immunohistochemically evaluated in 43 paraffin-embedded MFH specimens. Furthermore, the correlation of the TGF-beta and receptor expression with tumor proliferative activity assessed by MIB-1 indices was analyzed. RESULTS: Positive immunoreactivity for TGF-beta1, -beta2, and -beta 3 was identified in tumor cells of 42, 40, and 38 of the 43 MFHs, respectively. In each TGF-beta isoform immunostaining, the specimens were divided into two groups based on the number of positive tumor cells: those with low (<25%) and those with high (>==25%) immunoreactivity. There were no statistically significant differences in the MIB-1 indices between the two groups. Positive immunoreactivity for TGF-beta R1 and -beta R2 was identified in tumor cells of 36 and 24 of the MFHs, respectively. The specimens were divided into two groups based on their receptor expression patterns: those with both TGF-beta R1- and beta R2-positive immunoreactivity (n = 23), and those with both or either TGF beta R1- and -beta R2-negative immunoreactivity (n = 20). The MIB-1 indices in the both-TGF-beta R1- and -beta R2-positive group were significantly higher than those in the other group (P = 0.0102). There was no significant difference in pulmonary metastasis ratios between the two groups. CONCLUSIONS: These findings strongly suggest an association of the TGF-beta ligand/receptor system with a significantly higher MIB-1 index in human MFHs. Investigation of the TGF-beta R1 and -beta R2 coexpression might be useful in predicting tumor behavior of MFHs. PMID- 15355910 TI - Prognostic value of combined "triple"-reverse transcription-PCR analysis for prostate-specific antigen, human kallikrein 2, and prostate-specific membrane antigen mRNA in peripheral blood and lymph nodes of prostate cancer patients. AB - PURPOSE: We present the largest study of both peripheral blood and lymph node samples examining the utility of reverse transcription-polymerase chain reaction (RT-PCR) for established molecular markers as a diagnostic tool in the molecular staging of prostate cancer patients undergoing radical prostatectomy. EXPERIMENTAL DESIGN: Peripheral blood from 358 patients was obtained before radical prostatectomy. Corresponding obturatory lymph node samples were collected from 153 of these patients. Nested RT-PCR for prostate-specific antigen (PSA), human kallikrein 2 (hK2), and prostate-specific membrane antigen (PSMA) were performed on cDNA from peripheral blood. The lymph node cDNA was analyzed for PSA und hK2 expression. RESULTS: RT-PCR in peripheral blood was positive in 124 (34.6%) of 358 samples for PSA, 215 (60.1%) of 358 for PSMA, and 97 (27.1%) of 358 for hK2. Comparison of positive RT-PCR rates of pT(2) and pT(3) tumors in corresponding peripheral blood for PSA, PSMA, and hK2 were 31.9 and 40.0%, 58.8 and 62.5%, and 26.9 and 27.5%, respectively. Histopathologically, cancer-free lymph node samples were positive in RT-PCR for PSA and hK2 in 70 (49.6%) of 141 and 89 (63.2%) of 141 of cases. All histologically positive lymph node samples (n = 12, pN+) were positive for PSA RT-PCR. PSA RT-PCR alone, as well as combined PSA/PSMA RT-PCR evaluation, in peripheral blood showed a significant association with grading. PSA RT-PCR lymph node-negative samples were significantly less likely positive in their corresponding peripheral blood RT-PCR sample. CONCLUSIONS: Although the preoperative PSA RT-PCR in peripheral blood correlated with the grading of prostate cancer, no combination of RT-PCR results using "triple" markers (PSA, hK2, PSMA) in peripheral blood and/or lymph nodes yielded additional preoperative staging information. PMID- 15355911 TI - Tumor neoangiogenesis by CD31 and CD105 expression evaluation in breast carcinoma tissue microarrays. AB - PURPOSE: The aim of the study was to evaluate CD31 and CD105 immunohistochemical expressions in tissue microarrays from 360 breast carcinomas. STUDY DESIGN: Computerized (ACIS/Chromavision) assisted image analysis was performed to compare immunoreactions in tissue microarrays with those in current paraffin and frozen sections. We also aimed to determine the CD105 and CD31 prognostic significance and relevance in routine practice by correlating results of immunodetections with patients' (n = 360) outcome (14.3-year follow-up). RESULTS: The results show (a) that in tissue microarrays, the CD31 and CD105 expression quantified by image analysis device did not correlate with the measurements assessed on routine paraffin sections; (b) that CD105 expression is endowed of a prognostic significance in paraffin sections in terms of overall survival (P < 0.01), whereas in contrast, CD31 on paraffin sections did not correlate with patients overall survival; (c) that semiquantitative analysis of CD105 expression correlated with the image analysis measurements in frozen sections (rho = 0.671, P < 0.01) and paraffin (rho = 0.824, P < 0.01) sections. However, paraffin sections were less immunostained than frozen ones. CONCLUSIONS: It is concluded (a) that CD105 may be suitable in paraffin sections to evaluated neoangiogenesis; and (b) that tissue microarrays are not suitable substrates for neoangiogenesis evaluation as a prognostic indicator in breast carcinomas, in contrast to current tissue sections. PMID- 15355912 TI - Overexpressed eIF4E is functionally active in surgical margins of head and neck cancer patients via activation of the Akt/mammalian target of rapamycin pathway. AB - PURPOSE: Overexpression of eIF4E in surgical margins of head and neck cancer patients is an independent risk factor for recurrence. We hypothesize that overexpressed eIF4E is functionally active in tumor margins through activation of the Akt/mammalian target of rapamycin (mTOR) pathway EXPERIMENTAL DESIGN: Western blots and/or immunohistochemistry were performed to determine whether phosphorylation of mTOR and activation of its downstream molecules eIF4E-binding protein-1 (4E-BP1) and p70 S6 kinase and the upstream modulator of mTOR, Akt, were expressed in margins overexpressing eIF4E. RESULTS: There was a significant association between phospho-4E-BP1 and eIF4E expression of a margin or a significant difference in phospho-4E-BP1 expression between the eIF4E-positive and -negative margins (P < 0.01). A significant association between eIF4E and phospho-p70 S6 kinase as well as eIF4E and phospho-mTOR was also noted (P < 0.05). Western blot analysis indicated a highly significant difference in the phosphorylation status of 4E-BP1 between tumors and resection margins. A total of 89% of the 4E-BP1-expressing margins expressed more of the phosphorylated (beta, gamma, and delta) isoforms, whereas 81% of the 4E-BP1-expressing tumors expressed more of the unphosphorylated alpha isoform. A similar difference in Akt activation was noted between eIF4E-positive margins and tumors (P < 0.05). CONCLUSIONS: Overexpression of eIF4E is functionally active in tumor margins through activation of the Akt/mTOR signaling pathway. The greater degree of expression of downstream targets and upstream regulators of mTOR in margins compared with the tumors indicates preferential activation of the Akt/mTOR signaling pathway in margins overexpressing eIF4E. Rapamycin analogs can potentially be used as adjuvant therapy for patients with eIF4E-positive margins. PMID- 15355913 TI - Two proliferation-related proteins, TYMS and PGK1, could be new cytotoxic T lymphocyte-directed tumor-associated antigens of HLA-A2+ colon cancer. AB - PURPOSE: The purpose of this work was to provide a scientific basis for specific immunotherapy of colon cancer. EXPERIMENTAL DESIGN: This study focused on identification of colon tumor-associated antigens and HLA-A2-restricted and tumor reactive cytotoxic T lymphocytes (CTLs) generated from tumor-infiltrating lymphocytes of a colon cancer patient. A gene expression cloning method was used to identify genes coding for tumor antigens. Fifty-six peptides with HLA-A2 binding motifs encoded by these proteins were examined for their ability to induce HLA-A2-restricted and tumor-reactive CTLs. RESULTS: We identified the following three genes coding for proliferation-related proteins: thymidylate synthase (TYMS), which is involved in chemoresistance (5-fluorouracil); 5' aminoimidazole-4-carboxamide-1-beta-d-ribonucleotide transfolmylase/inosinicase (AICRT/I); and phosphoglycerate kinase 1 (PKG1), which was secreted by tumor cells and involved in the angiogenic process. TYMS was preferentially expressed in tumor cells, whereas AICRT/I and PKG1 were equally expressed in both cancer cells and normal tissues at the mRNA level. Among 56 peptides with HLA-A2-binding motifs encoded by these proteins, 8 peptides were recognized by the CTLs, and 5 of 8 peptides were also recognized by the CTL precursors without ex vivo activation in the peripheral blood of colon cancer patients. Furthermore, four of them (one each from TYMS and PKG1 and two from AICRT/1) possessed the ability to induce HLA-A2-restricted and peptide-specific CTLs cytotoxic to colon tumor cells in peripheral blood mononuclear cells of colon cancer patients. CONCLUSIONS: TYMS and PGK1, as well as their epitope peptides, might be appropriate target molecules for specific immunotherapy of HLA-A2(+) colon cancer patients because of the positive role of TYMS and PGK1 in chemoresistance (5-fluorouracil) and angiogenesis of tumor cells, respectively. PMID- 15355914 TI - Favorable neuroblastoma genes and molecular therapeutics of neuroblastoma. AB - PURPOSE AND EXPERIMENTAL DESIGN: Neuroblastoma (NB) is a common pediatric solid tumor that exhibits a striking clinical bipolarity: favorable and unfavorable. Favorable NB genes (EPHB6, EFNB2, EFNB3, NTRK1, and CD44) are genes whose high level expression predicts favorable NB outcome, and forced expression of these genes inhibits growth of unfavorable NB cells. In this study, we investigated whether favorable NB gene expression could be augmented in unfavorable NB cells by chemical compounds and whether an increased expression of these genes was associated with suppression of NB growth and metastasis. RESULTS: We found that inhibitors of DNA methylation [5-aza-2'-deoxycytidine (5AdC)], histone deacetylase (HDAC) [4-phenylbutyrate (4PB)], and proteasome (MG262) enhanced the expression of favorable NB genes in NB cell lines and inhibited the growth of these cells in vitro (P < 0.0005). The growth-inhibitory effects of 5AdC and 4PB in vitro were in part due to caspase-dependent cell death and inhibition of DNA synthesis. Administration of 5AdC and/or 4PB also suppressed growth of subcutaneous NB xenografts in nude mice (P < 0.001), which was accompanied by enhanced favorable NB gene expression and an increase in apoptosis. Moreover, 4PB suppressed bone marrow and liver metastases of NB cells in severe combined immunodeficient/Beige mice (P = 0.007 and P = 0.008, respectively). The growth suppressive activity of HDAC inhibitors on NB was further confirmed by the efficacy of trichostatin A, a potent and specific HDAC inhibitor. CONCLUSIONS: Collectively, these observations further emphasize the link between the elevated favorable NB gene expression and a benign phenotype of NB. PMID- 15355915 TI - p53 polymorphism and age of onset of hereditary nonpolyposis colorectal cancer in a Caucasian population. AB - PURPOSE: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant syndrome of familial malignancies. Colorectal and endometrial cancers are most frequently observed. The syndrome results mainly from germ-line mutations in DNA mismatch repair genes. A common G-to-C polymorphism at codon 72 in the p53 gene has been associated with increased risk for lung, nasopharyngeal, oral, prostate, and breast cancers and may be a marker for genetic susceptibility to colorectal cancer. We studied the influence of this p53 polymorphism on HNPCC age of onset. EXPERIMENTAL DESIGN: We determined the p53 genotype of 92 Caucasian mismatch repair mutation carriers, of which, 47 had colorectal cancer. The subjects were genotyped by single-strand conformational polymorphism analysis. We tested the association between age of onset and the p53 genotypes by comparing Kaplan-Meier survival curves, evaluating the homogeneity of the curves using the log-rank test and Wilcoxon's test, and estimating the association using the Cox proportional hazards regression model to adjust for potential demographic confounding factors. RESULTS: The HNPCC patients who were heterozygous developed their colorectal cancer 13 years earlier than HNPCC patients who were homozygous for the wild-type allele. CONCLUSIONS: Combining knowledge of an individual's p53 genotype with information on other genetic and environmental risk factors may improve risk estimates and help to identify individuals who are genetically susceptible to developing HNPCC at an earlier age. PMID- 15355916 TI - Steroid sulfatase and estrogen sulfotransferase in human endometrial carcinoma. AB - PURPOSE: Intratumoral metabolism and synthesis of estrogens are considered to play important roles in the pathogenesis and/or development of human endometrial carcinoma. Steroid sulfatase hydrolyzes biologically inactive estrogen sulfates to active estrogens, whereas estrogen sulfotransferase sulfonates estrogens to estrogen sulfates. However, the status of steroid sulfatase and/or estrogen sulfotransferase in human endometrial carcinoma has not been examined. EXPERIMENTAL DESIGN: We first examined the expression of steroid sulfatase and estrogen sulfotransferase in 6 normal endometrium and 76 endometrial carcinoma using immunohistochemistry to elucidate the possible involvement of steroid sulfatase and estrogen sulfotransferase. We then evaluated the enzymatic activity and the semiquantitative analysis of mRNA using reverse transcription-PCR in 21 endometrial carcinomas. We correlated these findings with various clinicopathological parameters including the expression of aromatase, 17beta hydroxysteroid dehydrogenase type 1 and type 2. RESULTS: Steroid sulfatase and estrogen sulfotransferase immunoreactivity was detected in 65 of 76 (86%) and 22 of 76 (29%) cases, respectively. Results of immunoreactivity for steroid sulfatase and estrogen sulfotransferase were significantly correlated with those of enzymatic activity and semiquantitative analysis of mRNA. No significant correlations were detected among the expression of the enzymes involved in intratumoral estrogen metabolism. There was a significant correlation between steroid sulfatase/estrogen sulfotransferase ratio and clinical outcomes of the patients. However, there were no significant differences between steroid sulfatase or estrogen sulfotransferase and estrogen receptor, progesterone receptor, Ki67, histologic grade, or clinical outcomes of the patients. CONCLUSIONS: Results of our study demonstrated that increased steroid sulfatase and decreased estrogen sulfotransferase expression in human endometrial carcinomas may result in increased availability of biologically active estrogens and may be related to estrogen-dependent biological features of carcinoma. PMID- 15355917 TI - SEL1L and squamous cell carcinoma of the esophagus. AB - The gene SEL1L is involved both in human breast and pancreatic cancer progression. It is located on 14q24.3-31, a region known to be lost in invasive cancer of the esophagus. We aimed to assess whether SEL1L could become a useful biomarker for this cancer. We assessed SEL1L mRNA and protein expression in 35 patients and found it to be weak in low-grade and strong in high-grade dysplasia. Although the majority of cancer patients showed differential expression (mRNA and protein) of SEL1L, in five cases it was completely absent; these patients had the worst outcomes. SEL1L immunoreactivity was negative in normal tissue samples from five patients with mild esophagitis as well as in normal mucosa adjacent to the tumor. We hypothesize that SEL1L could influence those cellular changes that mediate the transition from a normal mucosa to a neoplastic lesion and may help in the identification of those patients at higher risk of developing this cancer. The specific impact of SEL1L in esophageal cancer needs further investigation. PMID- 15355918 TI - Characterization of a human homologue of proteolysis-inducing factor and its role in cancer cachexia. AB - Cachexia is an important cause of secondary morbidity and mortality in patients with cancer. Previous studies have suggested that cancer-associated cachexia may be due in part to tumor-specific production and secretion of a glycosylated peptide, proteolysis-inducing factor, originally identified in a murine cancer cachexia model. We report here the cloning of a human cDNA that generates a peptide having high-sequence homology to this proteolysis-inducing factor. Constitutive expression of human proteolysis-inducing factor is low or absent in most normal human tissues but appears to be elevated in some human tumors. Stable forced expression of human proteolysis-inducing factor in multiple murine and human cell lines results in a secreted protein, but no glycosylation of the protein is detected. In addition, tumor xenografts engineered to overexpress human proteolysis-inducing factor protein do not induce cachexia in vivo. These findings raise important questions as to potential cross-species differences in protein sequence and processing of murine proteolysis-inducing factor and human proteolysis-inducing factor, as well as the nature of the relationship between human proteolysis-inducing factor and the development of cancer cachexia. PMID- 15355919 TI - Subepithelial myofibroblasts express cyclooxygenase-2 in colorectal tubular adenomas. AB - PURPOSE: Recent data support the hypothesis that the inducible isoform of cyclooxygenase (COX-2) plays a role in the early stages of colonic carcinogenesis and that nonsteroidal anti-inflammatory drugs (NSAIDs) retard the development of colon cancer by modulating COX-2. However, the cell types responsible for producing COX-2 in colorectal adenomas remain a subject of controversy. EXPERIMENTAL DESIGN: COX-2 expression in normal colonic mucosa (n = 50), hyperplastic polyps (n = 43), sporadic adenomas (n = 67), and invasive colonic adenocarcinoma (n = 39) was studied in formalin-fixed and paraffin-embedded tissue sections from endoscopy biopsy and colonic resection specimens. Immunohistochemistry (avidin-biotin complex technique with double immunolabeling) was used to identify the phenotypes of COX-2-producing cells. RESULTS: In colorectal adenomas, increased expression of COX-2 was detected and localized to alpha smooth muscle actin ( proportional, variant SMA)-positive subepithelial stromal cells (myofibroblasts) in the periluminal region of the lamina propria in 63 (94%) of 67 cases. In contrast, in normal colonic mucosa and in hyperplastic polyps with intact epithelium, COX-2 expression was found only in macrophages and endothelial cells. In areas in which the surface epithelium was ulcerated in normal mucosa as well as hyperplastic or neoplastic polyps, COX-2 expression was increased in granulation tissue (and present in macrophages, endothelium, and myofibroblasts). In invasive carcinoma, COX-2 expression in myofibroblasts was limited to the adenomatous portion of the tumor and was detected in 62% of cases (n = 39). In addition, focal expression of COX-2 by malignant epithelial cells was observed in 23% of invasive adenocarcinoma. CONCLUSIONS: These results show that increased COX-2 expression in sporadic adenoma of the colon is common and is localized specifically to subepithelial intestinal myofibroblasts. These findings further support the hypothesis that myofibroblasts are important target cells for NSAID-mediated chemoprevention of colorectal cancer. PMID- 15355920 TI - Thymidylate synthase gene polymorphism predicts toxicity in colorectal cancer patients receiving 5-fluorouracil-based chemotherapy. AB - PURPOSE: The target enzyme for 5-fluorouracil (5-FU) is thymidylate synthase (TS). The TYMS gene encoding this enzyme is polymorphic, having either double (2R) or tri-tandem (3R) repeats of a 28-bp sequence in the promoter region and a 6-bp variation in the 3'-untranslated region (3'-UTR). TS expression predicts response to 5-FU-based chemotherapy, and the expression seems to be determined by the TYMS gene promoter. The aim of this study was to investigate the utility of determining these two TYMS gene polymorphisms to predict the toxicity and efficacy of 5-FU treatment in patients with colorectal cancer. EXPERIMENTAL DESIGN: The determination of TYMS genotypes was performed in tumor and normal tissues by PCR amplification from 90 patients with colorectal cancer who were treated with adjuvant or palliative 5-FU-based chemotherapy. Associations between polymorphisms in the TYMS promoter and in the 3'-UTR gene and clinical outcome of these 90 patients treated with 5-FU based chemotherapy were evaluated individually. The linkage between TYMS promoter and TYMS 3'-UTR region polymorphisms was evaluated and a haplotype analysis was performed. RESULTS: Individuals who were homozygous for the double repeat in the TYMS promoter region had more severe side effects to 5-FU. Patients with a 2R/2R, a 2R/3R, or a 3R/3R genotype had a grade 3 or 4 toxicity rate of 43, 18, and 3% respectively (P < 0.01). The TYMS promoter and TYMS 3'-UTR polymorphisms were in linkage disequilibrium, and the haplotype 2R/ins 6-bp was significantly associated with a high risk of severe side effects to 5-FU. The TYMS promoter and TYMS 3'-UTR polymorphisms were not associated with a response to 5-FU and survival of patients who received palliative 5-FU-based chemotherapy. CONCLUSIONS: This study demonstrated that TYMS genotyping could be of help in predicting toxicity to 5-FU based chemotherapy. TYMS genotyping might make it possible to individualize treatment for patients with colorectal cancer. PMID- 15355921 TI - ABCG2 pharmacogenetics: ethnic differences in allele frequency and assessment of influence on irinotecan disposition. AB - PURPOSE: The ATP-binding cassette transporter ABCG2 (breast cancer resistance protein) is an efflux protein that plays a role in host detoxification of various xenobiotic substrates, including the irinotecan metabolite 7- ethyl-10 hydroxycamptothecin (SN-38). The ABCG2 421C>A polymorphism has been associated with reduced protein expression and altered function in vitro. The aim of this study was to evaluate the ethnic distribution and potential functional consequence of the ABCG2 421C>A genotype in cancer patients treated with irinotecan. EXPERIMENTAL DESIGN: ABCG2 genotyping was performed using Pyrosequencing on DNA from 88 American Caucasians, 94 African Americans, 938 Africans, and 95 Han Chinese, as well as in 84 European Caucasian patients treated with irinotecan undergoing additional blood sampling for pharmacokinetic studies. RESULTS: Significant differences in allele frequencies were observed between the given world populations (P < 0.001), the variant allele being most common in the Han Chinese population with a frequency as high as 34%. The mean area under the curve of irinotecan and SN-38 were 19,851 and 639 ng x hour/mL, respectively. The frequency of the variant allele (10.7%) was in line with results in American Caucasians. No significant changes in irinotecan pharmacokinetics were observed in relation to the ABCG2 421C>A genotype, although one of two homozygous variant allele carriers showed extensive accumulation of SN 38 and SN-38 glucuronide. CONCLUSIONS: The ABCG2 421C>A polymorphism appears to play a limited role in the disposition of irinotecan in European Caucasians. It is likely that the contribution of this genetic variant is obscured by a functional role of other polymorphic proteins. PMID- 15355922 TI - Elevated expression of A3 adenosine receptors in human colorectal cancer is reflected in peripheral blood cells. AB - PURPOSE: Adenosine is a ubiquitous nucleoside that accumulates at high levels in hypoxic regions of solid tumors, and A(3) adenosine receptors have been recently demonstrated to play a pivotal role in the adenosine-mediated inhibition of tumor cell proliferation. In the present work, we addressed the question of the putative relevance of A(3) subtypes in colorectal adenocarcinomas. EXPERIMENTAL DESIGN: Seventy-three paired samples of tumor and surrounding peritumoral normal mucosa at a distance of 2 and 10 cm from the tumor and blood samples obtained from a cohort of 30 patients with colorectal cancer were investigated to determine the presence of A(3) receptors by means of binding, immunocytochemistry, and real-time reverse transcription-polymerase chain reaction studies. RESULTS: As measured by receptor binding assays, the density of A(3) receptor was higher in colon carcinomas as compared with normal mucosa originating from the same individuals (P < 0.05). Overexpression of A(3) receptors at the protein level was confirmed by immunohistochemical studies, whereas no changes in A(3) mRNA accumulation in tumors as compared with the corresponding normal tissue were revealed. The overexpression of A(3) receptors in tumors was reflected in peripheral blood cells, where the density was approximately 3-fold higher compared with healthy subjects (P < 0.01). In a cohort of 10 patients studied longitudinally, expression of A(3) receptors in circulating blood cells returned to normal after surgical resection for colorectal cancer. CONCLUSIONS: This study provides the first evidence that A(3) receptor plays a role in colon tumorigenesis and, more importantly, can potentially be used as a diagnostic marker or a therapeutic target for colon cancer. PMID- 15355923 TI - Phospho-serine-118 estrogen receptor-alpha expression is associated with better disease outcome in women treated with tamoxifen. AB - PURPOSE: The purpose of this research was to determine whether estrogen receptor alpha specifically phosphorylated at Ser(118) is associated with clinical outcome in primary breast tumors from estrogen receptor-positive and node-negative breast cancer patients. EXPERIMENTAL DESIGN: Estrogen receptor alpha specifically phosphorylated at Ser(118) was determined by immunohistochemistry in 117 primary breast tumors from node-negative patients who were subsequently treated with adjuvant tamoxifen. The relationship of estrogen receptor alpha specifically phosphorylated at Ser(118) expression to disease-free survival and overall survival was determined. RESULTS: Estrogen receptor alpha specifically phosphorylated at Ser(118) was limited to estrogen receptor alpha ligand binding assay-positive tumors and among this subset was expressed in 70 (62%) of these tumors. Estrogen receptor alpha specifically phosphorylated at Ser(118) expression was more frequently observed in progesterone receptor-positive tumors compared with progesterone receptor-negative tumors (chi(2) test, P = 0.012, n = 113). A significant correlation was also seen between estrogen receptor alpha specifically phosphorylated at Ser(118) and progesterone receptor levels (Spearman r = 0.236, P = 0.0118, n = 113). Kaplan-Meier outcome analysis showed that patients whose primary tumors expressed estrogen receptor alpha specifically phosphorylated at Ser(118) had a longer disease-free survival (P = 0.0018, n = 113) and a trend toward better overall survival, but this was not statistically significant. Among the subset of progesterone receptor-positive tumors, progesterone receptor-positive/estrogen receptor alpha specifically phosphorylated at Ser(118)-positive patients had a significantly longer disease free survival that progesterone receptor-positive/estrogen receptor alpha specifically phosphorylated at Ser(118)-negative patients (P = 0.0041). CONCLUSIONS: Our data suggest that estrogen receptor alpha specifically phosphorylated at Ser(118) is a marker of a functional, intact ligand-dependent estrogen receptor signaling pathway in breast cancer and that estrogen receptor alpha specifically phosphorylated at Ser(118) status has the potential to provide a more precise biomarker of responsiveness to endocrine therapy in conjunction with estrogen receptor alpha and progesterone receptor status. PMID- 15355924 TI - Soluble type II transforming growth factor-beta receptor inhibits established murine malignant mesothelioma tumor growth by augmenting host antitumor immunity. AB - PURPOSE: Transforming growth factor (TGF)-beta blockade has been proposed as an anticancer therapy; however, understanding which tumor patients might benefit most from such therapy is crucial. An ideal target of such inhibitory therapy might be malignant mesothelioma (MM), a highly lethal, treatment-resistant malignancy of mesothelial cells of the pleura and peritoneum that produces large amounts of TGF-beta. The purpose of this study was to explore the possible therapeutic utility of TGF-beta blockade on MM. EXPERIMENTAL DESIGN: To evaluate this hypothesis, we tested the effects of a soluble TGF-beta type II receptor (sTGF-beta R) that specifically inhibits TGF-beta1 and TGF-beta 3 in three different murine MM tumor models, AB12 and AC29 (which produce large amounts of TGF-beta) and AB1 (which does not produce TGF-beta). RESULTS: Tumor growth of both established AB12 and AC29 tumors was inhibited by sTGF-beta R. In contrast, AB1 tumors showed little response to sTGF-beta R. The mechanism of these antitumor effects was evaluated and determined to be primarily dependent on immune-mediated responses because (a) the antitumor effects were markedly diminished in severe combined immunodeficient mice or mice depleted of CD8(+) T cells and (b) CD8(+) T cells isolated from spleens of mice treated with sTGF-beta R showed strong antitumor cytolytic effects, whereas CD8(+) T cells isolated from spleens of tumor-bearing mice treated with of control IgG2a showed no antitumor cytolytic effects. CONCLUSIONS: Our data suggest that TGF-beta blockade of established TGF-beta-secreting MM should be explored as a promising strategy to treat patients with MM and other tumors that produce TGF-beta. PMID- 15355925 TI - The role of hyperthermia in regional alkylating agent chemotherapy. AB - The role of hyperthermia during regional alkylating agent chemotherapy is controversial. The aim of this study was to determine the exact contribution of hyperthermia to tumor response during isolated limb infusion with l-phenylalanine mustard. Rats bearing rodent fibrosarcoma on the hindlimb underwent isolated limb infusion with saline, saline plus heat, l-phenylalanine mustard, l-phenylalanine mustard under conditions of normothermia, or l-phenylalanine mustard plus hyperthermia. Heat was administered locally using an in-line hot water circulation loop. Treatment with l-phenylalanine mustard at a concentration of 15 or 50 micrograms/mL was ineffective at producing tumor growth delay (P = 0.24 and 0.41, respectively). Furthermore, thermal enhancement of l-phenylalanine mustard activity was not seen at 15 micrograms/mL. However, administration of high-dose l phenylalanine mustard, 50 micrograms/mL, with increasing amounts of heat yielded increasing tumor growth delay, increased regressions, and decreased proliferative index. Although l-phenylalanine mustard infusion under normothermia yielded a tumor growth delay of 7.1 days, combination l-phenylalanine mustard + hyperthermia treatment produced tumor growth delay of 27.0 days (P < 0.01; with two of five animals showing a complete response). Four hours after isolated limb infusion, 50.9% of cells in tumor treated with l-phenylalanine mustard + hyperthermia experienced apoptosis, whereas only 18.1, 16, and 4.4% of cells underwent apoptosis after treatment with l-phenylalanine mustard, saline + hyperthermia, or saline. The mean concentration of l-phenylalanine mustard within tumor relative to perfusate following isolated limb infusion was found to be similar among all groups at 0.023, 0.025, and 0.032 in animals undergoing isolated limb infusion with l-phenylalanine mustard, l-phenylalanine mustard + normothermia, and l-phenylalanine mustard + hyperthermia, respectively. These data indicate a synergistic cytotoxic effect of l-phenylalanine mustard + hyperthermia in isolated limb infusion, which is not attributable to enhanced tumor drug uptake. PMID- 15355926 TI - Simultaneously targeting epidermal growth factor receptor tyrosine kinase and cyclooxygenase-2, an efficient approach to inhibition of squamous cell carcinoma of the head and neck. AB - PURPOSE: Epidermal growth factor receptor (EGFR) and cyclooxygenase-2 (Cox-2) contribute to development of squamous cell carcinoma of the head and neck (SCCHN). Simultaneously blocking both EGFR and Cox-2-mediated pathways may be an efficient means of inhibiting cancer cell growth in SCCHN. EXPERIMENTAL DESIGN: A combination of EGFR-selective tyrosine kinase inhibitors (TKIs) AG1478 or ZD1839 (Iressa or gefitinib) with a Cox-2 inhibitor (Cox-2I) celecoxib (Celebrex) was studied for its effects on cell growth, cell cycle progression, and apoptosis in SCCHN cell lines by cell growth assay, clonogenic assay, flow cytometric analysis, and terminal deoxynucleotidyl transferase-mediated nick end labeling assay. A potential effect of EGFR TKIs and Cox-2I on angiogenesis was examined by endothelial capillary tube formation assay. Primary and secondary targets of EGFR TKIs and Cox-2I were also examined using immunoblotting and immunoprecipitation after the combined treatment. RESULTS: The combination of AG1478 or ZD1839 with celecoxib either additively or synergistically inhibited growth of the five SCCHN cell lines examined, significantly induced G(1) arrest and apoptosis, and suppressed capillary formation of endothelium. Furthermore, the combination showed strong reductions of p-EGFR, p-extracellular signal-regulated kinase 1/2, and p-Akt in SCCHN cells as compared with the single agents. Both AG1478 and ZD1839 inhibited expression of Cox-2 protein, whereas celecoxib mainly blocked the production of prostaglandin E(2). CONCLUSIONS: These results suggest that cell growth inhibition induced by a combination of EGFR TKIs and Cox-2I is mediated through simultaneously blocking EGFR and Cox-2 pathways. This combination holds a great potential for the treatment and/or prevention of SCCHN. PMID- 15355927 TI - Antitumor activity of gefitinib in malignant rhabdoid tumor cells in vitro and in vivo. AB - PURPOSE: Malignant rhabdoid tumor (MRT) is a rare and highly aggressive neoplasm of young children. Current treatments have had only limited success. Epidermal growth factor receptor (EGFR) was found recently to be expressed on MRT cell lines. Gefitinib (trade name Iressa) is an oral and selective EGFR-tyrosine kinase inhibitor and has been demonstrated to be effective in inhibiting the proliferation of cancer cells in vivo as well as in clinical trials. This encouraged us to examine the antitumor effects of gefitinib on MRT cells in vitro and in vivo. EXPERIMENTAL DESIGN: The expression of EGFR in two MRT tumors and two MRT cell lines (MP-MRT-AN and KP-MRT-NS), established from these two tumor tissues, was examined by immunohistochemistry, immunofluorescence, and immunoblot. The effect of gefitinib on EGFR phosphorylation was examined by immunoblot. The effects of gefitinib on cell growth and apoptosis were examined by cell growth assay and terminal deoxynucleotidyl transferase-mediated nick end labeling assay. The in vivo effect of gefitinib was assessed in athymic mice that had been xenografted with MRT cells. RESULTS: The expression of EGFR was detected in both tumor tissues and cell lines. Gefitinib inhibited EGFR-phosphorylation (IC(50) < 0.1 micromol/L) and in vitro cell growth (IC(50) = approximately 10-12 micromol/L), and a high concentration of gefitinib (20 micromol/L) induced apoptosis in vitro (MP-MRT-AN, 42.9% and KP-MRT-NS, 47.2%). Furthermore, gefitinib at 150 mg/kg had a cytostatic effect on established MRT xenografts (MP MRT-AN, P = 0.039 and 0.0014; and KP-MRT-NS, P = 0.048 and 0.0086). CONCLUSIONS: Our results demonstrate that gefitinib has antitumor effects in MRT cells in vitro and in vivo and, thus, has promise as a novel and therapeutic strategy for MRT. PMID- 15355928 TI - Thalidomide pharmacokinetics and metabolite formation in mice, rabbits, and multiple myeloma patients. AB - PURPOSE: Thalidomide has a variety of biological effects that vary considerably according to the species tested. We sought to establish whether differences in pharmacokinetics could form a basis for the species-specific effects of thalidomide. EXPERIMENTAL DESIGN: Mice and rabbits were administered thalidomide (2 mg/kg) p.o. or i.v., and plasma concentrations of thalidomide were measured after drug administration using high performance liquid chromotography. Plasma samples from five multiple myeloma patients over 24 hours after their first dose of thalidomide (200 mg) were similarly analyzed and all data were fitted to a one compartment model. Metabolites of thalidomide in plasma were identified simultaneously using liquid chromatography-mass spectrometry. RESULTS: Plasma concentration-time profiles for the individual patients were very similar to each other, but widely different pharmacokinetic properties were found between patients compared with those in mice or rabbits. Area under the concentration curve values for mice, rabbits, and multiple myeloma patients were 4, 8, and 81 micromol/L. hour, respectively, and corresponding elimination half-lives were 0.5, 2.2, and 7.3 hours, respectively. Large differences were also observed between the metabolite profiles from the three species. Hydrolysis products were detected for all species, and the proportion of hydroxylated metabolites was higher in mice than in rabbits and undetectable in patients. CONCLUSIONS: Our results show major interspecies differences in the pharmacokinetics of thalidomide that are related to the altered degree of metabolism. We suggest that the interspecies differences in biological effects of thalidomide may be attributable, at least in part, to the differences in its metabolism and hence pharmacokinetics. PMID- 15355929 TI - In vitro toxicity of A-431 carcinoma cells with antibodies to epidermal growth factor receptor and epithelial glycoprotein-1 conjugated to radionuclides emitting low-energy electrons. AB - PURPOSE: The ability of antibodies (Abs) conjugated to radionuclides emitting low energy electrons to specifically kill nonadherent lymphoma target cells in vitro was demonstrated previously. This study extends this work to adherent carcinoma cells. The fact that these cells are spread out on plastic can potentially make it more difficult to deliver radiation to the nucleus from decays in the cytoplasm or on the cell surface. EXPERIMENTAL DESIGN: The Abs tested were anti epidermal growth factor receptor and anti-epithelial glycoprotein-1, conjugated to indium-111 or iodine-125, which emit low-energy Auger and conversion electrons. Conjugates of the beta-particle emitter, iodine-131, also were tested, for comparison. Abs were incubated with the cells for 2 days, and then the treated cells were assayed for colony-forming units. The radiation dose delivered to the nucleus was calculated from the cumulative decays per cell. RESULTS: With conjugates of (111)In, very potent killing was obtained with both of the Abs, with 100% kill (approximately 4-5 logs) even at subsaturating Ab concentrations. Lower levels of kill were obtained with (125)I or (131)I conjugates. Conjugates with (131)I, a beta-particle emitter, produced greater nonspecific toxicity. The greater potency of (111)In could be attributed to the higher specific activity that was obtained routinely with this radiolabel, up to 70 mCi/mg. Uptake of radioactivity peaked at approximately 200 cpm per cell. Dosimetry calculations, using subcellular S values, demonstrated that the toxicity observed was consistent with the amount of radiation delivered to the nucleus. CONCLUSIONS: These results are similar to previous results obtained with B lymphoma cells and indicate that this approach is applicable to a wide range of tumor types. Radionuclides emitting low-energy electrons are effective at killing target cells with relatively little nonspecific toxicity, if sufficient activity is delivered to the cell. Most Abs to high-density cell surface antigens would probably be effective. PMID- 15355930 TI - Consideration of QT/QTc interval data in a phase I study in patients with advanced cancer. PMID- 15355931 TI - Limitations of the adenoma-carcinoma sequence in colorectum. PMID- 15355932 TI - Sinorhizobium meliloti strain 1021 produces a low-molecular-mass capsular polysaccharide that is a homopolymer of 3-deoxy-D-manno-oct-2-ulosonic acid harboring a phospholipid anchor. AB - Sinorhizobium meliloti strain 1021 possesses the particularity to synthesize biologically inefficient capsular polysaccharides (KPS). It has been assumed that this class of compounds is not produced in high-molecular-mass (HMM) forms, even if many genetic analyses show the existence of expression of genes involved in the biosynthesis of capsular polysaccharides. The expression of these genes that are involved in the export of a KPS throughout the membrane and in the attachment of a lipid moiety has never been related to a structurally characterized surface polysaccharide. It is now reported that S. meliloti strain 1021 produces low molecular-mass polysaccharides (4-4.5 kDa) that are exclusively composed of beta (2-->7)-linked 3-deoxy-d-manno-oct-2-ulopyranosonic acid (Kdo) residues. These compounds are considered precursor molecules of HMM KPS, whose biosynthesis is arrested in the case of S. meliloti strain 1021. For the first time, the phospholipid anchor of a rhizobial KPS has been found, and its structure could be partially identified-namely, a phosphoglycerol moiety bearing a hydroxy octacosanoic acid. When compared to other rhizobial KPS (composed of dimeric hexose-Kdo-like sugar repeating units), the Kdo homopolymer described here may explain why a complementation of S. meliloti strain 1021 Exo B mutant with an effective rkpZ gene restoring an active higher KPS size does not completely lead to the fully effective nitrogen fixing phenotype. PMID- 15355933 TI - RANTES (CCL5) induces a CCR5-dependent accelerated shedding of syndecan-1 (CD138) and syndecan-4 from HeLa cells and forms complexes with the shed ectodomains of these proteoglycans as well as with those of CD44. AB - We recently demonstrated that RANTES forms complexes with CCR5, syndecan-1 (SD 1), SD-4, and CD44 expressed by human primary macrophages and that SD-1 and SD-4 but neither CD44 nor SD-2 coimmunoprecipitate with CCR5. Here we show that RANTES directly binds in a glycosaminoglycan-dependent manner to SD-1, SD-4, and CD44. Moreover, RANTES accelerates the shedding of SD-1 and SD-4 ectodomains from HeLa cells expressing CCR5 and, by contrast, has no effect on the constitutive shedding of CD44 from these cells. These accelerated sheddings are prevented by the MEK1/2 inhibitor, U0126, and by the protein kinase C inhibitor bisindolylmaleimide I. This indicates that both MAP kinase--and protein kinase C dependent signaling pathways are involved in these RANTES-induced accelerated sheddings. RANTES also induces a decreased expression of SD-1 and SD-4 by HeLa cells expressing CCR5 and on the contrary an increased expression of CD44 by these cells. By contrast, RANTES neither accelerates the shedding of SD-1 and SD 4 ectodomains from HeLa cells lacking CCR5, nor changes the SD-1-, SD-4-, and CD44-plasma membrane expressions of these cells. CCR5 is therefore involved in the RANTES-induced accelerated shedding of SD-1 and SD-4 ectodomains. Nevertheless, the fact that RANTES stimulates in Hela cells (expressing or lacking CCR5) the mRNA synthesis of SD-1 and SD-4 indicates that the molecular events that follow the synthesis of these proteoglycans differ, according to the presence or not of CCR5. Finally, RANTES forms GAG-dependent complexes with the shed ectodomains of SD-1 and SD-4 as well as with those of CD44. The role of these events in the pathophysiology of RANTES deserves further study. PMID- 15355937 TI - Trial registration: a great idea switches from ignored to irresistible. PMID- 15355936 TI - Clinical trial registration: a statement from the International Committee of Medical Journal Editors. PMID- 15355938 TI - Pseudomonas aeruginosa-induced infection and degradation of human wound fluid and skin proteins ex vivo are eradicated by a synthetic cationic polymer. AB - OBJECTIVES: Antimicrobial peptides are important effectors of innate immunity. Bacteria display multiple defence mechanisms against these peptides. For example, Pseudomonas aeruginosa releases potent proteinases that inactivate the human cathelicidin LL-37. Hence, in conditions characterized by persistent bacterial colonization, such as in P. aeruginosa-infected skin wounds, there is a need for efficient means of reducing bacterial load. Here, the effect of the cationic molecule polyhexamethylenebiguanide (PHMB) was evaluated. METHODS: Infection models in human wound fluid and human skin were established. Radial diffusion methods, bacterial growth and bactericidal assays were used for determination of effects of PHMB on bacteria in the presence of plasma, wound fluid or human skin. At the protein and tissue levels, SDS-PAGE, light microscopy and scanning electron microscopy were used to study the effects of P. aeruginosa infection before and after addition of PHMB. RESULTS: PHMB killed common ulcer-derived bacteria in the presence of human wound fluid. Furthermore, elastase-expressing P. aeruginosa completely degraded wound fluid proteins as well as human skin during infection ex vivo. The infection, and consequent protein degradation, was reversed by PHMB. CONCLUSIONS: The ex vivo infection models presented here should be helpful in the screening of novel antimicrobials and constitute a prerequisite for future clinical studies. PMID- 15355939 TI - Effect of n-octanesulphonylacetamide (OSA) on ATP and protein expression in Mycobacterium bovis BCG. AB - OBJECTIVE: To determine the effect on BCG of n-octanesulphonylacetamide (OSA), a novel compound of the class beta-sulphonylcarboxamides, which has potent in vitro activity against pathogenic mycobacteria. METHODS AND RESULTS: The effect of OSA in BCG was examined using two-dimensional protein electrophoresis. Treatment of BCG with OSA resulted in overexpression of two proteins identified as the b subunit of ATP synthase (Rv1306) and a 17 kDa heat shock protein (Rv0251c). [35S]Methionine pulse-labelling revealed that overexpression occurred within as little as 3.5 h post-exposure. These results were confirmed by RT-PCR. ATP levels decreased in OSA-treated BCG at 5 min, and 1, 3 and 24 h, with a 64%, 45%, 54% and 73% reduction in ATP, respectively. Only dicyclohexylcarbodiimide (DCCD), a known ATP synthase inhibitor, had a similar effect. No appreciable difference in ATP level was observed in BCG treated with standard antimycobacterial drugs, additional respiratory chain inhibitors or a fatty acid synthase inhibitor at a comparable time-point. Protein synthesis decreased within 5 min of exposure to OSA (56%), DCCD (74%) and thenoyltrifluoroacetone (TTFA) (77%). Ethanol (2.3%) potentiated the activity of OSA. In contrast, no synergic effect was observed with streptomycin and ethanol. Mycolic acid levels decreased 79% with DCCD, 46% with TTFA, a complex II inhibitor, and 43% with OSA compared with untreated controls. CONCLUSIONS: Our results suggest that OSA may interfere directly or indirectly with ATP synthase and possibly other components of the mycobacterial respiratory chain. These effects may hinder energy production, leading to interruption in the synthesis of large macromolecules including proteins and mycolic acids. PMID- 15355940 TI - Growth as a part of the composite endpoint in paediatric antiretroviral clinical trials. AB - Paediatric HIV is a rapidly emerging disease in many resource-poor countries. Survival into adulthood is possible for HIV-infected children provided that they receive effective antiretroviral therapy (ART). Large trials comparing multiple regimens of ART in children of resource-poor countries have not been completed. Design of those trials will need to incorporate both lessons learned from trials completed in developed countries as well as unique aspects of the developing countries in which they are conducted. Trial design will necessarily include close attention to the endpoint, and in children, special consideration will need to be given to growth as a component of the endpoint--whether or not growth should be a component of the composite endpoint, and if so, how. PMID- 15355941 TI - Community-acquired methicillin-resistant Staphylococcus aureus arrives in Hong Kong. PMID- 15355942 TI - Selection of topoisomerase mutations and overexpression of adeB mRNA transcripts during an outbreak of Acinetobacter baumannii. AB - OBJECTIVES: To investigate the mechanism of ciprofloxacin resistance in isolates of Acinetobacter baumannii during two hospital outbreaks and to determine the expression level of the gene encoding the AdeB efflux pump. METHODS: Isolates were previously typed by PFGE and their MICs determined by broth microdilution. The gyrA and parC genes were sequenced and the adeB gene examined by real-time reverse transcription PCR (RT-PCR). RESULTS: Two clonal lineages were responsible for the two hospital outbreaks. In both outbreaks, ciprofloxacin susceptibility was reduced during the course of the outbreak when compared with the index isolates. Mutations in gyrA and parC were found to have occurred during the outbreak. The MICs of non-fluoroquinolone antibiotics were raised in one clonal lineage and this was associated with a >10-fold increase in mRNA transcripts for adeB. CONCLUSIONS: We have witnessed the appearance of gyrA and parC mutations during outbreaks of A. baumannii. In parallel with these mutations, we observed up-regulation of the adeB gene associated with a decrease in susceptibility to non-fluoroquinolone antibiotics. These data illustrate the propensity for A. baumannii to develop multi-drug resistance rapidly. PMID- 15355943 TI - The prevalence, predictors, and consequences of peripheral sensory neuropathy in older patients. AB - BACKGROUND: The prevalence, predictors, and consequences of peripheral neuropathy in the elderly have not been well defined. METHODS: Seven hundred ninety-five noninstitutionalized patients 65 years of age and older, recruited from the practices of family physicians, completed questionnaires and underwent peripheral neurologic examinations and tests of gait and balance. Variables included sociodemographic information, medical conditions, symptoms (numbness, pain, trouble with balance or walking, and restless legs), quality of life measures, ankle reflexes, position sense, vibratory sense, fine touch sensation, Tinnetti balance examination, and a 50-foot timed walk. RESULTS: The prevalence of at least one bilateral sensory deficit rose from 26% for 65- to 74-year-olds to 54% for those 85 and older. The most common deficit was loss of ankle reflex followed by loss of fine touch. Only 40% of those with bilateral deficits reported having a disease known to cause peripheral neuropathy. Predictors of bilateral deficits included increasing age, income less than 15,000 dollars, a history of military service, increasing body mass index, self-reported history of diabetes mellitus, Vitamin B12 deficiency or rheumatoid arthritis, and absence of a history of hypertension. Deficits were associated with numbness, pain, restless legs, trouble walking, trouble with balance, and reduced quality of life. CONCLUSIONS: Peripheral sensory deficits are common in the elderly. In most cases, a medical cause is not obvious. Their consequences may not be as benign as often supposed. PMID- 15355944 TI - Patients' understanding and use of snack food package nutrition labels. AB - PURPOSE: Little information exists on how patients in medical practice use food package nutrition labels. We theorized that patients in a general medical practice might not make the distinction between serving size and total package nutrition information, and this might lead to obesity. METHODS: Ninety patients between ages of 18 and 65 attending the St. Francis/University of Tennessee Family Practice Center were interviewed to determine whether they could calculate the total calories in sample snack food packages that contained more than one serving. RESULTS: Ninety percent of our patient sample correctly identified the number of calories per individual serving, but only 37% were able to recognize that the sample packages contained multiple servings. Confusion between calories per serving size and total calories per package was correlated with lower educational levels (P =.011) and with the presence of cardiovascular heart disease in our patient sample. CONCLUSIONS: Our patients tended to think of a multiple serving package as one serving. They underestimated and under-reported caloric intake from snack food sources. We conclude that snack food labels as actually used by patients do not lead to informed dietary choices. These findings could impact our understanding and management of the obesity epidemic in the United States. PMID- 15355945 TI - Parents' health and demographic characteristics predict noncompliance with well child visits. AB - BACKGROUND: The purpose of this study was to investigate factors related to well child visit noncompliance in an ethnically diverse family practice clinic population. METHODS: Participants included 146 parents (131 mothers and 15 fathers) of children aged 0 to 24 months who received care at a St. Paul residency clinic. Participants completed telephone surveys that asked about their demographic characteristics, attitudes toward well-child visits, whether the most recent planned well-child visit had been kept, and their own and their child's health characteristics. RESULTS: All participants thought that well-child visits were important, with immunizations being the highest rated reason for importance. Fourteen percent of parents said they had missed a recent well-child visit, mostly because they forgot. More than three fourths of parents believed visit reminders were helpful, and the preferred type of reminder was a telephone call. Noncompliance with well-child visits was associated with the parent's depressive symptoms, transportation difficulties, working at a job, having private (vs public) health insurance, and being older (vs younger). CONCLUSIONS: These results suggest that well-child visit compliance might be enhanced by visit reminders and improved access to transportation. The relationship of well-child visit noncompliance to parental depressive symptoms, if verified in other populations, points to a need for greater surveillance of children/families who do not schedule or keep well-child visits. PMID- 15355946 TI - Intimate partner violence: what are physicians' perceptions? AB - BACKGROUND: Intimate partner violence (IPV) is common in primary care; 11% to 22% of women experienced physical abuse in the past year. Older women experience IPV as well, but it is often undetected. This study examined primary care providers' awareness about IPV in older women, including their screening practices and management. METHODS: Interviews and focus groups were conducted with 44 primary care providers. Thematic analysis was used to identify common themes. RESULTS: Providers fell along a continuum of thoroughness for identifying and managing IPV in older women, ranging from suboptimal to thorough identification of IPV and suboptimal to thorough management of the patient. In addition to the barriers commonly reported about IPV screening in younger women, providers described limited understanding of the diagnoses commonly associated with IPV, frustration with older women's unwillingness to disclose problems and ask for help, and limited community services that accommodate older women with IPV. Providers recommended that communities sponsor public awareness campaigns about IPV as a problem for all women and that aging and IPV agencies work together. CONCLUSIONS: Continued provider training about IPV should include information on identifying older victims and appropriate management options. Participants stressed the importance of community efforts to raise awareness and improve resources available for older women who are victims of IPV. PMID- 15355947 TI - Continuity of care and patient satisfaction in a family practice clinic. AB - BACKGROUND: Continuity is a tenant central to family practice. Continuity is associated with improved satisfaction in populations that can easily change providers. However, little is known about the importance of continuity where patients are assigned providers. METHODS: A pretested survey was distributed to patients of a family practice residency clinic in a military medical center for a week's period. Results were analyzed using chi2, unpaired t test, correlation matrices, and linear regression for patient satisfaction. RESULTS: The response rate was 68.3%. Responders were not more likely to be seeing their primary care provider (PCP). Regression analysis revealed that 12% of patient satisfaction was associated with long-term continuity rates, 23% by PCP satisfaction, and 17% by how easy it was to make the appointment. For high clinic users (>10 visits/year), 78% of patient satisfaction is determined by PCP satisfaction and long-term continuity rates. A subset of patients (13%) values choice of appointment time or other providers over PCP continuity. Satisfaction is not diminished in this group despite low long-term continuity (P <.05 for all results). CONCLUSIONS: Patient satisfaction is associated with continuity, especially for high clinic users. Although continuity is important, a subset of patients values the ability to see other providers and to change providers. PMID- 15355948 TI - Management of brown recluse spider bites in primary care. AB - BACKGROUND: Treatment of brown recluse spider bites remains controversial; there are multiple options but little evidence of their effectiveness. METHODS: Over a 5-year period, family physicians enrolled consecutive patients with suspected brown recluse spider bites. Usual care was provided based on physician preferences. Topical nitroglycerine patches and vitamin C tablets were provided at no cost for those who wished to use them. Baseline data were collected, and patients were followed-up weekly until healing occurred. Outcome measures included time to healing and occurrence of scarring. Regression methods were used to evaluate the impact of the 4 main treatment approaches (corticosteroids, dapsone, topical nitroglycerine, and high-dose vitamin C) after controlling for bite severity and other predictors. RESULTS: Two hundred and sixty-two patients were enrolled; outcomes were available for 189. The median healing time was 17 days. Only 21% had permanent scarring. One hundred seventy-four received a single treatment modality. Among this group, 12 different modalities were used. After controlling for other variables, predictors of more rapid healing included lower severity level, less erythema, and less necrosis at time of presentation, younger age, no diabetes, and earlier medical attention. Systemic corticosteroids and dapsone were associated with slower healing. Predictors of scarring were higher severity, presence of necrosis, and diabetes. Dapsone was associated with an increased probability of scarring. CONCLUSIONS: We found no evidence that commonly used treatment approaches reduced healing time or the likelihood of scarring in suspected brown recluse spider bites. PMID- 15355949 TI - Colonoscopy in rural communities: can family physicians perform the procedure with safe and efficacious results? AB - BACKGROUND: Colonoscopy is becoming increasingly necessary for many patients in screening, diagnosing, and treating colorectal problems. Because the majority of rural doctors are family physicians, providing colonoscopy for the enlarging group of patients with valid indications in rural areas is difficult, unless rural family physicians perform the procedure. Subspecialists in academic settings have been responsible for most of the previously reported studies regarding colonoscopy. We have studied the safety and efficacy of the procedure when performed by rural family physicians. METHODS: A total of 200 sequential colonoscopies performed by family physicians in a rural setting were prospectively collected. Outcomes were measured based on current recommendations and benchmarks, including rate of reaching the cecum, time to reach the cecum, time to completion of the study, pathologic lesions found, and complications. RESULTS: The rate of reaching the cecum was 96.5%, and the average time to the cecum was 15.9 minutes. The average time to study completion was 34.4 minutes. The rate of neoplastic polyps and cancer found was 22.5% and 2.5%, respectively. There were no serious complications. CONCLUSIONS: Adequately trained family physicians can provide safe and technically competent colonoscopy in a rural setting. Their results compare favorably to the currently reported comparative benchmarks from other endoscopists. PMID- 15355950 TI - Diagnosis and treatment of obesity in adults: an applied evidence-based review. AB - BACKGROUND: Obesity is epidemic and leads to substantial morbidity/mortality. Effective strategies exist for managing obesity yet are rarely used by physicians. This applied evidence-based review provides a rationale for the diagnosis and treatment of obesity in adults by providing test characteristics for the body mass index (BMI) and number needed to treat (NNT) for relevant treatments. METHODS: We integrated evidence supporting recommendations from scientific bodies addressing obesity in adults, including: the National Heart, Lung, and Blood Institute, the World Health Organization, the Canadian Task Force on Preventive Health Care, and the US Preventive Task Force. In addition, pertinent studies were identified from MEDLINE, Database of Abstracts of Reviews of Effectiveness, and the Cochrane Database. RESULTS: (1) manage obesity as a chronic relapsing disease; (2) use BMI as a vital sign to screen for overweight/obese patients and to decide treatment (positive predictive value of 97%); (3) modest weight loss (10%) positively affects prevention/treatment of hypertension (NNT = 3), diabetes (NNT = 9), and hyperlipidemia; (4) effective treatments exist for overweight/obese patients and a combination of diet and exercise provides the best results (NNT = 7); (5) counsel patients to achieve a goal of 10% reduction in weight (500 to 800 kcal/day decrease to affect 1- to 2 pound loss/week); (6) counsel patients to exercise to achieve a goal of any increased energy expenditure. CONCLUSIONS: Weight loss has an impact on important disease states and risk factors. Effective strategies exist for the management of obesity when viewed as a chronic relapsing disease. PMID- 15355951 TI - Religion, spirituality, and the practice of medicine. AB - Physicians are confronted with new information from the popular media, peer reviewed journals, and their patients regarding the association of religious and spiritual factors with health outcomes. Although religion and spirituality have become more visible within health care, there are considerable ethical issues raised when physicians incorporate these dimensions into their care. Spiritualities are responsive to patient needs by offering beliefs, stories, and practices that facilitate the creation of a personally meaningful world, a constructed "reality" in the face of illness, disability, or death. It is largely through narrative that physicians incorporate into the health care encounter the spiritualities that are central to their patients' lived experience of illness and health. PMID- 15355952 TI - Follow-up on family practice residents' perspectives on length and content of training. AB - BACKGROUND: The structure of family practice residency programs remains essentially unchanged from the model first proposed more than 35 years ago. Advances in medical technology and knowledge combined with increasing restrictions on resident work hours and decreasing medical student interest invite reconsideration of how family physicians are trained. METHODS: We resurveyed 442 third-year family practice residents who had participated in a prior study in 2000 to determine whether their opinions about the length and content of residency had changed and whether they would still choose to be a physician and a family physician. RESULTS: Thirty-seven percent of responding third-year residents favored extending family practice residency to 4 years. Compared as groups, there was relatively little change in opinion between first- and third-year residents. However, residents' individual responses about the settings and content areas for which they would be willing to consider extending training varied considerably between years 1 and 3. Personal characteristics did not seem to influence residents' opinions about length and content of training. Reasons for favoring a 4-year program and barriers to change were similar to those reported previously. Residents' commitment to medicine and family medicine was still strong and was not associated with their opinions about length of training. CONCLUSION: Although most surveyed residents favored a 3-year residency program, a substantial minority still supported extending training to 4 years, and the majority would still choose to enter family medicine programs if they were extended. Given a lack of consensus about specific content areas, family medicine should consider a period of experimentation to determine how to best prepare future family physicians. PMID- 15355953 TI - Who provides care to Medicare beneficiaries and what settings do they use? AB - BACKGROUND: Concerns have been expressed that the physician workforce is unprepared for the explosion in the number of older persons in America. As a step toward informing these discussions, this article will describe how Medicare beneficiaries currently access physician services. METHODS: This study is a descriptive analysis of the physician services used by Medicare beneficiaries. The Medicare Beneficiary Survey (MCBS) data from 1998 are used for the analysis. The locations of service delivery were compared among family physicians, general internists, and geriatricians. RESULTS: The physician office was the most common site of service, comprising 49% of all provider claims. General internists (20.1% of office claims) and family physicians (18.6% of office claims) were the most common providers. Family physicians spent the largest proportion of their time in the office (77%), general internists were the most likely provider to see patients in the hospital (19%), and geriatricians were the most likely to see patients in the nursing home (27%) and at home (1.7%). CONCLUSIONS: Office-based care by general internists and family physicians constitute a major infrastructure element in the delivery of care to Medicare beneficiaries. The practices of geriatric medicine physicians are more heavily weighted toward the nursing home setting. PMID- 15355954 TI - High-dose fentanyl patch for cancer pain. AB - OBJECTIVE: To describe a successful experience with a high dose (1000 microg/hr) of transdermal fentanyl for cancer pain relief. CASE REPORT: A 62-year-old man suffering from rectal carcinoma was treated by our home care hospice unit during his last 3.5 months of life. At admission to our home care unit, he suffered mostly from severe anal pain (verbal pain scale of 10/10) due to advanced disease. He was then on 150 microg/hr transdermal fentanyl. Adjuvant therapy with amitriptyline 50 mg/day and dexamethasone 4 mg/day was added, but it did not relieve his pain. The dose of transdermal fentanyl was increased gradually to 1000 microg/hr with good pain control (verbal pain scale of 1 to 4/10 most of the time). Before his death, he was mentally alert with good pain control. CONCLUSIONS: High doses of transdermal fentanyl (1000 microg/hr; 10 patches) should be considered for pain relief in cancer patients. PMID- 15355955 TI - How long does it take to become a competent family physician? PMID- 15355956 TI - Using a bed sheet to avoid an assisted delivery. PMID- 15355957 TI - The variable location, content, and legibility of expiration dates on medicine containers. PMID- 15355958 TI - Human light chain 3/MAP1LC3B is cleaved at its carboxyl-terminal Met121 to expose Gly120 for lipidation and targeting to autophagosomal membranes. AB - Human light chain 3/MAP1LC3B, an autophagosomal ortholog of yeast Atg8, is conjugated to phospholipid (PL) via ubiquitylation-like reactions mediated by human Atg7 and Atg3. Since human Atg4B was found to cleave the carboxyl terminus of MAP1LC3B in vitro, we hypothesized that this exposes its carboxyl-terminal Gly(120). It was recently reported, however, that when Myc-MAP1LC3B-His is expressed in HEK293 cells, its carboxyl terminus is not cleaved. (Tanida, I., Sou, Y.-s., Ezaki, J., Minematsu-Ikeguchi, N., Ueno, T., and Kominami, E. (2004) J. Biol. Chem. 279, 36268-36276). To clarify this contradiction, we sought to determine whether the carboxyl terminus of MAP1LC3B is cleaved to expose Gly(120) for further ubiquitylation-like reactions. When MAP1LC3B-3xFLAG and Myc-MAP1LC3B His were expressed in HEK293 cells, their carboxyl termini were cleaved, whereas there was little cleavage of mutant proteins MAP1LC3B(G120A)-3xFLAG and Myc MAP1LC3B(G120A)-His, containing Ala in place of Gly(120). An in vitro assay showed that Gly(120) is essential for carboxyl-terminal cleavage by human Atg4B as well as for formation of the intermediates Atg7-MAP1LC3B (ubiquitin-activating enzyme-substrate) and Atg3-MAP1LC3B (ubiquitin carrier protein-substrate). Recombinant MAP1LC3B-PL was fractionated into the 100,000 x g pellet in a manner similar to that shown for endogenous MAP1LC3B-PL. RNA interference of MAP1LC3B mRNA resulted in a decrease in both endogenous MAP1LC3B-PL and MAP1LC3B. These results indicate that the carboxyl terminus of MAP1LC3B is cleaved to expose Gly(120) for further ubiquitylation-like reactions. PMID- 15355959 TI - Structural basis of cellular redox regulation by human TRP14. AB - Thioredoxin-related protein 14 (TRP14) is involved in regulating tumor necrosis factor-alpha-induced signaling pathways in a different manner from human thioredoxin 1 (Trx1). Here, we report the crystal structure of human TRP14 determined at 1.8-A resolutions. The structure reveals a typical thioredoxin fold with characteristic structural features that account for the substrate specificity of the protein. The surface of TRP14 in the vicinity of the active site includes an extended loop and an additional alpha-helix, and the distribution of charged residues in the surface is different from Trx1. The distinctive dipeptide between the redox-active cysteines contributes to stabilizing the thiolate anion of the active site cysteine 43, increasing reactivity of the cysteine toward substrates. These structural differences in the active site suggest that TRP14 has evolved to regulate cellular redox signaling by recognizing a distinctive group of substrates that would complement the group of proteins regulated by Trx1. PMID- 15355960 TI - In vivo light-induced and basal phospholipase C activity in Drosophila photoreceptors measured with genetically targeted phosphatidylinositol 4,5 bisphosphate-sensitive ion channels (Kir2.1). AB - The phosphatidylinositol 4,5-bisphosphate (PIP(2))-sensitive inward rectifier channel Kir2.1 was expressed in Drosophila photoreceptors and used to monitor in vivo PIP(2) levels. Since the wild-type (WT) Kir2.1 channel appeared to be saturated by the prevailing PIP(2) concentration, we made a single amino acid substitution (R228Q), which reduced the effective affinity for PIP(2) and yielded channels generating currents proportional to the PIP(2) levels relevant for phototransduction. To isolate Kir2.1 currents, recordings were made from mutants lacking both classes of light-sensitive transient receptor potential channels (TRP and TRPL). Light resulted in the effective depletion of PIP(2) by phospholipase C (PLC) in approximately three or four microvilli per absorbed photon at rates exceeding approximately 150% of total microvillar phosphoinositides per second. PIP(2) was resynthesized with a half-time of approximately 50 s. When PIP(2) resynthesis was prevented by depriving the cell of ATP, the Kir current spontaneously decayed at maximal rates representing a loss of approximately 40% loss of total PIP(2) per minute. This loss was attributed primarily to basal PLC activity, because it was greatly decreased in norpA mutants lacking PLC. We tried to confirm this by using the PLC inhibitor U73122; however, this was found to act as a novel inhibitor of the Kir2.1 channel. PIP(2) levels were reduced approximately 5-fold in the diacylglycerol kinase mutant (rdgA), but basal PLC activity was still pronounced, consistent with the suggestion that raised diacylglycerol levels are responsible for the constitutive TRP channel activity characteristic of this mutant. PMID- 15355961 TI - Annexin 6 modulates the maxi-chloride channel of the apical membrane of syncytiotrophoblast isolated from human placenta. AB - The syncytiotrophoblast separates the maternal and fetal blood and constitutes the primary barrier for maternal-fetal transport. The Maxi-chloride channel from the apical membrane of the syncytiotrophoblast plays a role in the chloride conductance. Annexins can play an important role in the regulation of membrane events. In this study we evaluate the role of annexin 6 in the Maxichloride channel properties. The results showed that annexin 6 is bound in the apical placenta membranes in a calcium-dependent phospholipid-binding manner but also in a calcium-independent fashion. The neutralization of annexin 6 decreased the total current by 39 +/- 1.9% in the range of +/-80 mV, and the currents decrease with the time. The single-channel slope conductance was decreased from 253 +/- 7.4 pS (control) to 105 +/- 13 pS, and the amplitude decreased by 50%. The open probability was also affected when higher voltage steps were used, changes in either the positive or negative direction induced the channel to close, and the open probability (P(o)) did not decrease. In channels with neutralized annexin 6, it was maintained at 1 at +/-40 mV and at +/-80 mV. These results suggest that endogenous annexin 6 could regulate the Maxi-chloride channel. The results obtained with normal placentae, in which annexin 6 was neutralized, are similar to those described for the Maxi-chloride channel isolated from pre-eclamptic placenta. Together these data suggest that annexin 6 could play an important role in ion transport of the placenta. PMID- 15355962 TI - Phosphorylation of IQGAP1 modulates its binding to Cdc42, revealing a new type of rho-GTPase regulator. AB - The Rho-GTPase Cdc42 is important for the establishment and maintenance of epithelial polarity. Signaling from Cdc42 is propagated via its effector molecules that specifically bind to Cdc42 in the GTP-bound form. The cell-cell contact regulator and actin-binding protein IQGAP1 is described as effector of Cdc42 and Rac. Unexpectedly, we show in this study that IQGAP1 bound also directly nucleotide-depleted Cdc42 (Cdc42-ND). This interaction was enhanced in the presence of phosphatase inhibitors and in epithelial cells without cell-cell contacts. Tandem mass spectrometry analysis and immunoprecipitation experiments revealed that IQGAP1 was Ser1443-phosphorylated in vivo, potentially by protein kinase Cepsilon and upon loss of cell-cell contacts. In addition, we identified two independent domains of the IQGAP1 C terminus that bound exclusively Cdc42-ND. These domains interacted with each other, favoring the binding to Cdc42-GTP. Moreover, phosphorylation on Ser1443 strongly inhibited this intramolecular interaction. Thus, we unraveled a molecular mechanism that reveals a novel type of Rho-GTPase regulator. We propose that, depending on its phosphorylation state, IQGAP1 might serve as an effector or sequester nucleotide-free Cdc42 to prevent signaling. PMID- 15355963 TI - Involvement of the nonhomologous region of subunit A of the yeast V-ATPase in coupling and in vivo dissociation. AB - The catalytic nucleotide binding subunit (subunit A) of the vacuolar proton translocating ATPase (or V-ATPase) is homologous to the beta-subunit of the F ATPase but contains a 90-amino acid insert not present in the beta-subunit, termed the nonhomologous region. We previously demonstrated that mutations in this region lead to changes in coupling of proton transport and ATPase activity and to inhibition of in vivo dissociation of the V-ATPase complex, an important regulatory mechanism (Shao, E., Nishi T., Kawasaki-Nishi, S., and Forgac, M. (2003) J. Biol. Chem. 278, 12985-12991). Measurement of the ATP dependence of coupling for the wild type and mutant proteins demonstrates that the coupling differences are observed at ATP concentrations up to 1 mm. A decrease in coupling efficiency is observed at higher ATP concentrations for the wild type and mutant V-ATPases. Immunoprecipitation of an epitope-tagged nonhomologous region from cell lysates indicates that this region is able to bind to the integral V0 domain in the absence of the remainder of the A subunit, an interaction confirmed by immunoprecipitation of V0. Interaction between the nonhomologous region and V0 is reduced upon incubation of cells in the absence of glucose, suggesting that the nonhomologous region may act as a trigger to activate in vivo dissociation. Immunoprecipitation suggests that the epitope tag on the nonhomologous region becomes less accessible upon glucose withdrawal, possibly due to binding to another cellular target. In vivo dissociation of the V-ATPase in response to glucose removal is also blocked by chloroquine, a weak base that neutralizes the acidic pH of the vacuole. The results suggest that the dependence of in vivo dissociation of the V-ATPase on catalytic activity may be due to neutralization of the yeast vacuole, which in turn blocks glucose-dependent dissociation. PMID- 15355964 TI - Mutation of the aromatic amino acid interacting with adenine moiety of ATP to a polar residue alters the properties of multidrug resistance protein 1. AB - Structural analyses of several bacterial ATP-binding cassette (ABC) transporters indicate that an aromatic amino acid residue in a nucleotide-binding domain (NBD) interacts with the adenine ring of the bound ATP and contributes to the ATP binding. Substitution of this aromatic residue with a polar serine residue in bacterial histidine transporter completely abolished both ATP binding and ATP dependent histidine transport. However, substitution of the aromatic amino acid residue in the human cystic fibrosis transmembrane conductance regulator with a polar cysteine residue did not have any effect on the ATP-dependent chloride channel function of the protein. To determine whether the other eucaryotic ABC transporters use the strategy analogous to that in some bacterial ABC transporters, the aromatic Trp653 residue in NBD1 and the Tyr1302 residue in NBD2 of human multidrug resistance-associated protein 1 (MRP1) was mutated to either a different aromatic residue or a polar cysteine residue. Substitution of the aromatic residue with a different aromatic amino acid, such as W653Y or Y1302W, did not affect ATP-dependent leukotriene C4 (LTC4) transport. In contrast, substitution of the aromatic residue with a polar cysteine residue, such as W653C or Y1302C, decreased the affinity for ATP, resulting in greatly increased Kd values for ATP binding or Km values for ATP in ATP-dependent LTC4 transport. Interestingly, although substitution of the aromatic Trp653 in NBD1 of MRP1 with a polar cysteine residue greatly decreases the affinity for ATP, the ATP dependent LTC4 transport activities are much higher than that of wild-type MRP1, supporting our hypothesis that the increased release rate of the bound ATP from the mutated NBD1 facilitates the protein to start a new cycle of ATP-dependent solute transport. PMID- 15355965 TI - Structural and dynamic independence of isopeptide-linked RanGAP1 and SUMO-1. AB - Although sumoylation regulates a diverse and growing number of recognized biological processes, the molecular mechanisms by which the covalent attachment of the ubiquitin-like protein SUMO can alter the properties of a target protein remain to be established. To address this question, we have used NMR spectroscopy to characterize the complex of mature SUMO-1 with the C-terminal domain of human RanGAP1. Based on amide chemical shift and 15N relaxation measurements, we show that the C terminus of SUMO-1 and the loop containing the consensus sumoylation site in RanGAP1 are both conformationally flexible. Furthermore, the overall structure and backbone dynamics of each protein remain unchanged upon the covalent linkage of Lys524 in RanGAP1 to the C-terminal Gly97 of SUMO-1. Therefore, SUMO-1 and RanGAP1 behave as "beads-on-a-string," connected by a flexible isopeptide tether. Accordingly, the sumoylation-dependent interaction of RanGAP1 with the nucleoporin RanBP2 may arise through the bipartite recognition of both RanGAP1 and SUMO-1 rather than through a new binding surface induced in either individual protein upon their covalent linkage. We hypothesize that this conformational flexibility may be a general feature contributing to the recognition of ubiquitin-like modified proteins by their downstream effector machineries. PMID- 15355966 TI - Structural and biochemical identification of a novel bacterial oxidoreductase. AB - By using a bioinformatics screen of the Escherichia coli genome for potential molybdenum-containing enzymes, we have identified a novel oxidoreductase conserved in the majority of Gram-negative bacteria. The identified operon encodes for a proposed heterodimer, YedYZ in Escherichia coli, consisting of a soluble catalytic subunit termed YedY, which is likely anchored to the membrane by a heme-containing trans-membrane subunit termed YedZ. YedY is uniquely characterized by the presence of one molybdenum molybdopterin not conjugated by an additional nucleotide, and it represents the only molybdoenzyme isolated from E. coli characterized by the presence of this cofactor form. We have further characterized the catalytic subunit YedY in both the molybdenum- and tungsten substituted forms by using crystallographic analysis. YedY is very distinct in overall architecture from all known bacterial reductases but does show some similarity with the catalytic domain of the eukaryotic chicken liver sulfite oxidase. However, the strictly conserved residues involved in the metal coordination sphere and in the substrate binding pocket of YedY are strikingly different from that of chicken liver sulfite oxidase, suggesting a catalytic activity more in keeping with a reductase than that of a sulfite oxidase. Preliminary kinetic analysis of YedY with a variety of substrates supports our proposal that YedY and its many orthologues may represent a new type of membrane associated bacterial reductase. PMID- 15355967 TI - Partitioning of NaPi cotransporter in cholesterol-, sphingomyelin-, and glycosphingolipid-enriched membrane domains modulates NaPi protein diffusion, clustering, and activity. AB - In dietary potassium deficiency there is a decrease in the transport activity of the type IIa sodium/phosphate cotransporter protein (NaPi) despite an increase in its apical membrane abundance. This novel posttranslational regulation of NaPi activity is mediated by the increased glycosphingolipid content of the potassium deficient apical membrane. However, the mechanisms by which these lipids modulate NaPi activity have not been determined. We determined if in potassium deficiency NaPi is increasingly partitioned in cholesterol-, sphingomyelin-, and glycosphingolipid-enriched microdomains of the apical membrane and if the increased presence of NaPi in these microdomains modulates its activity. By using a detergent-free density gradient flotation technique, we found that 80% of the apical membrane NaPi partitions into the low density cholesterol-, sphingomyelin , and GM1-enriched fractions characterized as "lipid raft" fractions. In potassium deficiency, a higher proportion of NaPi was localized in the lipid raft fractions. By combining fluorescence correlation spectroscopy and photon counting histogram methods for control and potassium-deficient apical membranes reconstituted into giant unilamellar vesicles, we showed a 2-fold decrease in lateral diffusion of NaPi protein and a greater than 2-fold increase in size of protein aggregates/clusters in potassium deficiency. Our results indicate that NaPi protein is localized in membrane microdomains, that in potassium deficiency a larger proportion of NaPi protein is present in these microdomains, and that NaPi lateral diffusion is slowed down and NaPi aggregation/clustering is increased in potassium deficiency, both of which could be associated with the decreased Na/Pi cotransport activity in potassium deficiency. PMID- 15355968 TI - Discovery and characterization of a novel, widely expressed metalloprotease, ADAMTS10, and its proteolytic activation. AB - We describe the discovery and characterization of ADAMTS10, a novel metalloprotease encoded by a locus on human chromosome 19 and mouse chromosome 17. ADAMTS10 has the typical modular organization of the ADAMTS family, with five thrombospondin type 1 repeats and a cysteine-rich PLAC (protease and lacunin) domain at the carboxyl terminus. Its domain organization and primary structure is similar to a novel long form of ADAMTS6. In contrast to many ADAMTS proteases, ADAMTS10 is widely expressed in adult tissues and throughout mouse embryo development. In situ hybridization analysis showed widespread expression of Adamts10 in the mouse embryo until 12.5 days of gestation, after which it is then expressed in a more restricted fashion, with especially strong expression in developing lung, bone, and craniofacial region. Mesenchymal, not epithelial, expression in the developing lung, kidney, gonad, salivary gland, and gastrointestinal tract is a consistent feature of Adamts10 regulation. N-terminal sequencing and treatment with decanoyl-Arg-Val-Lys-Arg-chloromethylketone indicate that the ADAMTS10 zymogen is processed by a subtilisin-like proprotein convertase at two sites (Arg64/Gly and Arg233/Ser). The widespread expression of ADAMTS10 suggests that furin, a ubiquitously expressed proprotein convertase, is the likely processing enzyme. ADAMTS10 expressed in HEK293F and COS-1 cells is N glycosylated and is secreted into the medium, as well as sequestered at the cell surface and extracellular matrix, as demonstrated by cell surface biotinylation and immunolocalization in nonpermeabilized cells. ADAMTS10 is a functional metalloprotease as demonstrated by cleavage of alpha2-macroglobulin, although physiological substrates are presently unknown. PMID- 15355969 TI - Elements in the N-terminal signaling sequence that determine cytosolic topology of short-chain dehydrogenases/reductases. Studies with retinol dehydrogenase type 1 and cis-retinol/androgen dehydrogenase type 1. AB - High affinity, retinoid-specific binding proteins chaperone retinoids to manage their transport and metabolism. Proposing mechanisms of retinoid transfer between these binding proteins and membrane-associated retinoid-metabolizing enzymes requires insight into enzyme topology. We therefore determined the topology of mouse retinol dehydrogenase type 1 (Rdh1) and cis-retinoid androgen dehydrogenase type 1 (Crad1) in the endoplasmic reticulum of intact mammalian cells. The properties of Rdh1 were compared with a chimera with a luminal signaling sequence (11beta-hydroxysteroid dehydrogenase (11beta-HSD1)(1-41)/Rdh1(23-317); the green fluorescent protein (GFP) fusion proteins Rdh1(1-22)/GFP, Crad1(1-22)/GFP, and 11beta-HSD1(1-41)/GFP; and signaling sequence charge difference mutants using confocal immunofluorescence, antibody access, proteinase K sensitivity, and deglycosylation assays. An N-terminal signaling sequence of 22 residues, consisting of a hydrophobic helix ending in a net positive charge, anchors Rdh1 and Crad1 in the endoplasmic reticulum facing the cytoplasm. Mutating arginine to glutamine in the signaling sequence did not affect topology. Inserting one or two arginine residues near the N terminus of the signaling sequence caused 28-95% inversion from cytoplasmic to luminal, depending on the net positive charge remaining at the C terminus of the signaling sequence; e.g. the mutant L3R,L5R,R16Q,R19Q,R21Q faced the lumen. Experiments with N- and C-terminal epitope-tagged Rdh1 and molecular modeling indicated that a hydrophobic helix turn-helix near the C terminus of Rdh1 (residues 289-311) projects into the cytoplasm. These data provide insight into the features necessary to orient type III (reverse signal-anchor) proteins and demonstrate that Rdh1, Crad1, and other short-chain dehydrogenases/reductases, which share similar N-terminal signaling sequences such as human Rdh5 and mouse Rdh4, orient with their catalytic domains facing the cytoplasm. PMID- 15355970 TI - Activity-dependent transcriptional activation and mRNA stabilization for cumulative expression of pituitary adenylate cyclase-activating polypeptide mRNA controlled by calcium and cAMP signals in neurons. AB - Although it has been established that an activity-dependent gene transcription is induced by the calcium (Ca(2+)) signals in neurons, it is unclear how the specific mRNA moieties are transiently accumulated in response to synaptic transmission which evokes multiple intracellular signals including Ca(2+) and cAMP ones. The expression of pituitary adenylate cyclase activating polypeptide (PACAP), a neuropeptide, is controlled by Ca(2+) signals evoked via membrane depolarization in neurons, and, in cultured rat cortical neuronal cells, we found that the Ca(2+) signal-mediated activation of the PACAP gene promoter was critically controlled by a single cAMP-response element (CRE) located at around 200, to which the CRE-binding protein predominantly bound. The Ca(2+) signal induced expression of PACAP mRNA was enhanced by forskolin, which evokes cAMP signals. In support, the PACAP gene promoter was synergistically enhanced by Ca(2+) and cAMP signals through the CRE, accompanying a prolonged activation of extracellular signal-related protein kinase 1/2 and CRE-binding protein. On the other hand, sole administration of forskolin markedly reduced the cellular content of PACAP mRNA, which was restored by the addition of Ca(2+) signals. We found that the stability of PACAP mRNA was increased in response to Ca(2+) signals but not that of activity-regulated cytoskeleton-associated protein (Arc) mRNA, indicating an activity-dependent stabilization of specific mRNA species in neurons, which can antagonize the regulation mediated by cAMP signals. Thus, the transcriptional activation and mRNA stabilization are coordinately regulated by Ca(2+) and cAMP signals for the cumulative expression of PACAP mRNA in neurons. PMID- 15355972 TI - Crystal structure of agmatinase reveals structural conservation and inhibition mechanism of the ureohydrolase superfamily. AB - Agmatine is the product of arginine decarboxylation and can be hydrolyzed by agmatinase to putrescine, the precursor for biosynthesis of higher polyamines, spermidine, and spermine. Besides being an intermediate in polyamine metabolism, recent findings indicate that agmatine may play important regulatory roles in mammals. Agmatinase is a binuclear manganese metalloenzyme and belongs to the ureohydrolase superfamily that includes arginase, formiminoglutamase, and proclavaminate amidinohydrolase. Compared with a wealth of structural information available for arginases, no three-dimensional structure of agmatinase has been reported. Agmatinase from Deinococcus radiodurans, a 304-residue protein, shows approximately 33% of sequence identity to human mitochondrial agmatinase. Here we report the crystal structure of D. radiodurans agmatinase in Mn(2+)-free, Mn(2+) bound, and Mn(2+)-inhibitor-bound forms, representing the first structure of agmatinase. It reveals the conservation as well as variation in folding, oligomerization, and the active site of the ureohydrolase superfamily. D. radiodurans agmatinase exists as a compact homohexamer of 32 symmetry. Its binuclear manganese cluster is highly similar but not identical to the clusters of arginase and proclavaminate amidinohydrolase. The structure of the inhibited complex reveals that inhibition by 1,6-diaminohexane arises from the displacement of the metal-bridging water. PMID- 15355971 TI - Caveolin-1 gene disruption promotes mammary tumorigenesis and dramatically enhances lung metastasis in vivo. Role of Cav-1 in cell invasiveness and matrix metalloproteinase (MMP-2/9) secretion. AB - Caveolin-1 (Cav-1) is the principal structural component of caveolae membrane domains in non-muscle cells, including mammary epithelia. There is now clear evidence that caveolin-1 influences the development of human cancers. For example, a dominant-negative mutation (P132L) in the Cav-1 gene has been detected in up to 16% of human breast cancer samples. However, the exact functional role of caveolin-1 remains controversial. Mechanistically, in cultured cell models, Cav-1 is known to function as a negative regulator of the Rasp42/44 MAP kinase cascade and as a transcriptional repressor of cyclin D1 gene expression, possibly explaining its in vitro transformation suppressor activity. Genetic validation of this hypothesis at the in vivo and whole organismal level has been prevented by the lack of a Cav-1 (-/-)-null mouse model. Here, we examined the role of caveolin-1 in mammary tumorigenesis and lung metastasis using a molecular genetic approach. We interbred a well characterized transgenic mouse model of breast cancer, MMTV-PyMT (mouse mammary tumor virus-polyoma middle T antigen), with Cav 1 (-/-)-null mice. Then, we followed the onset and progression of mammary tumors and lung metastases in female mice over a 14-week period. Interestingly, PyMT/Cav 1 (-/-) mice showed an accelerated onset of mammary tumors, with increased multiplicity and tumor burden ( approximately 2-fold). No significant differences were detected between PyMT/Cav-1 (+/+) and PyMT/Cav-1 (+/-) mice, indicating that complete loss of caveolin-1 is required to accelerate both tumorigenesis and metastasis. Molecularly, mammary tumor samples derived from PyMT/Cav-1 (-/-) mice showed ERK-1/2 hyperactivation, cyclin D1 up-regulation, and Rb hyperphosphorylation, consistent with dys-regulated cell proliferation. PyMT/Cav 1 (-/-) mice also developed markedly advanced metastatic lung disease. Conversely, recombinant expression of Cav-1 in a highly metastatic PyMT mammary carcinoma-derived cell line, namely Met-1 cells, suppressed lung metastasis by approximately 4.5-fold. In vitro, these Cav-1-expressing Met-1 cells (Met-1/Cav 1) demonstrated a approximately 4.8-fold reduction in invasion through Matrigel coated membranes. Interestingly, delivery of a cell permeable peptide encoding the caveolin-1 scaffolding domain (residues 82-101) into Met-1 cells was sufficient to inhibit invasion. Coincident with this decreased invasive index, Met-1/Cav-1 cells exhibited marked reductions in MMP-9 and MMP-2 secretion and associated gelatinolytic activity, as well as diminished ERK-1/2 signaling in response to growth factor stimulation. These results demonstrate, for the first time, that caveolin-1 is a potent suppressor of mammary tumor growth and metastasis using novel in vivo animal model approaches. PMID- 15355973 TI - Crystal structure of T-protein of the glycine cleavage system. Cofactor binding, insights into H-protein recognition, and molecular basis for understanding nonketotic hyperglycinemia. AB - The glycine cleavage system catalyzes the oxidative decarboxylation of glycine in bacteria and in mitochondria of animals and plants. Its deficiency in human causes nonketotic hyperglycinemia, an inborn error of glycine metabolism. T protein, one of the four components of the glycine cleavage system,is a tetrahydrofolate dependent aminomethyltransferase. It catalyzes the transfer of the methylene carbon unit to tetrahydrofolate from the methylamine group covalently attached to the lipoamide arm of H-protein. To gain insight into the T protein function at the molecular level, we have determined the first crystal structure of T-protein from Thermotoga maritima by the multiwavelength anomalous diffraction method of x-ray crystallography and refined four structures: the apoform; the tetrahydrofolate complex; the folinic acid complex; and the lipoic acid complex. The overall fold of T-protein is similar to that of the C-terminal tetrahydrofolate-binding region (residues 421-830) of Arthrobacter globiformis dimethylglycine oxidase. Tetrahydrofolate (or folinic acid) is bound near the center of the tripartite T-protein. Lipoic acid is bound adjacent to the tetrahydrofolate binding pocket, thus defining the interaction surface for H protein binding. A homology model of the human T-protein provides the structural framework for understanding the molecular mechanisms underlying the development of nonketotic hyperglycinemia due to missense mutations of the human T-protein. PMID- 15355974 TI - Crystal structure of avian aminoimidazole-4-carboxamide ribonucleotide transformylase in complex with a novel non-folate inhibitor identified by virtual ligand screening. AB - Aminoimidazole-4-carboxamide ribonucleotide transformylase (AICAR Tfase), one of the two folate-dependent enzymes in the de novo purine biosynthesis pathway, is a promising target for anti-neoplastic chemotherapy. Although classic antifolates, such as methotrexate, have been developed as anticancer agents, their general toxicity and drug resistance are major issues associated with their clinical use and future development. Identification of inhibitors with novel scaffolds could be an attractive alternative. We present here the crystal structure of avian AICAR Tfase complexed with the first non-folate based inhibitor identified through virtual ligand screening of the National Cancer Institute Diversity Set. The inhibitor 326203-A (2-[5-hydroxy-3-methyl-1-(2-methyl-4-sulfophenyl)-1H pyrazol-4-ylazo]-4-sulfo-benzoic acid) displayed competitive inhibition against the natural cofactor, 10-formyl-tetrahydrofolate, with a K(i) of 7.1 mum. The crystal structure of AICAR Tfase with 326203-A at 1.8 A resolution revealed a unique binding mode compared with antifolate inhibitors. The inhibitor also accessed an additional binding pocket that is not occupied by antifolates. The sulfonate group of 326203-A appears to form the dominant interaction of the inhibitor with the proposed oxyanion hole through interaction with a helix dipole and Lys(267). An aromatic interaction with Phe(316) also likely contributes to favorable binding. Based on these structural insights, several inhibitors with improved potency were subsequently identified in the National Cancer Institute Compound Library and the Available Chemical Directory by similarity search and molecular modeling methods. These results provide further support for our combined virtual ligand screening rational design approach for the discovery of novel, non-folate-based inhibitors of AICAR Tfase. PMID- 15355975 TI - Regulation of PTEN phosphorylation and stability by a tumor suppressor candidate protein. AB - The tumor suppressor PTEN plays an essential role in regulating signaling pathways involved in cell growth and apoptosis and is inactivated in a wide variety of tumors. In this study, we have identified a protein, referred to as PICT-1 (protein interacting with carboxyl terminus 1), that binds to the C terminus of PTEN and regulates its phosphorylation and turnover. Down-regulation of PICT-1 in MCF7 cells by RNA interference enhances the degradation of PTEN with a concomitant decrease in its phosphorylation. PTEN C-terminal tumor-associated mutants, which are highly susceptible to protein degradation, have lost the ability to bind to PICT-1 along with their reduced phosphorylation, suggesting that their rapid turnover results from impaired binding to PICT-1. Our results identify PICT-1 as a PTEN-interacting protein that promotes the phosphorylation and stability of PTEN. These findings suggest a novel molecular mechanism underlying the turnover of PTEN, which also provides an explanation for the loss of PTEN function due to C-terminal mutations. PMID- 15355976 TI - Recoverin and rhodopsin kinase activity in detergent-resistant membrane rafts from rod outer segments. AB - Cholesterol-rich membranes or detergent-resistant membranes (DRMs) have recently been isolated from bovine rod outer segments and were shown to contain several signaling proteins such as, for example, transducin and its effector, cGMP phosphodiesterase PDE6. Here we report the presence of rhodopsin kinase and recoverin in DRMs that were isolated in either light or dark conditions at high and low Ca2+ concentrations. Inhibition of rhodopsin kinase activity by recoverin was more effective in DRMs than in the initial rod outer segment membranes. Furthermore, the Ca2+ sensitivity of rhodopsin kinase inhibition in DRMs was shifted to lower free Ca2+ concentration in comparison with the initial rod outer segment membranes (IC50=0.76 microm in DRMs and 1.91 microm in rod outer segments). We relate this effect to the high cholesterol content of DRMs because manipulating the cholesterol content of rod outer segment membranes by methyl beta-cyclodextrin yielded a similar shift of the Ca2+-dependent dose-response curve of rhodopsin kinase inhibition. Furthermore, a high cholesterol content in the membranes also increased the ratio of the membrane-bound form of recoverin to its cytoplasmic free form. These data suggest that the Ca2+-dependent feedback loop that involves recoverin is spatially heterogeneous in the rod cell. PMID- 15355977 TI - The spindle pole body assembly component mps3p/nep98p functions in sister chromatid cohesion. AB - For successful chromosome segregation during mitosis, several processes must occur early in the cell cycle, including spindle pole duplication, DNA replication, and the establishment of cohesion between nascent sister chromatids. Spindle pole body duplication begins in G1 and continues during early S-phase as spindle pole bodies mature and start to separate. Key steps in spindle pole body duplication are the sequential recruitment of Cdc31p and Spc42p by the nuclear envelope transmembrane protein Msp3p/Nep98p (herein termed Mps3p). Concurrent with DNA replication, Ctf7p/Eco1p (herein termed Ctf7p) ensures that nascent sister chromatids are paired together, identifying the products of replication as sister chromatids. Here, we provide the first evidence that the nuclear envelope spindle pole body assembly component Mps3p performs a function critical to sister chromatid cohesion. Mps3p was identified as interacting with Ctf7p from a genome wide two-hybrid screen, and the physical interaction was confirmed by both in vivo (co-immunoprecipitation) and in vitro (GST pull-down) assays. An in vivo cohesion assay on new mps3/nep98 alleles revealed that loss of Mps3p results in precocious sister chromatid separation and that Mps3p functions after G1, coincident with Ctf7p. Mps3p is not required for cohesion during mitosis, revealing that Mps3p functions in cohesion establishment and not maintenance. Mutated Mps3p that results in cohesion defects no longer binds to Ctf7p in vitro, demonstrating that the interaction between Mps3p and Ctf7p is physiologically relevant. In support of this model, mps3 ctf7 double mutant cells exhibit conditional synthetic lethality. These findings document a new role for Mps3p in sister chromatid cohesion and provide novel insights into the mechanism by which a spindle pole body component, when mutated, contributes to aneuploidy. PMID- 15355978 TI - Adenomatous polyposis coli is down-regulated by the ubiquitin-proteasome pathway in a process facilitated by Axin. AB - Adenomatous polyposis coli (APC) protein and Axin form a complex that mediates the down-regulation of beta-catenin, a key effector of Wnt signaling. Truncation mutations in APC are responsible for familial and sporadic colorectal tumors due to failure in the down-regulation of beta-catenin. While the regulation of beta catenin by APC has been extensively studied, the regulation of APC itself has received little attention. Here we show that the level of APC is down-regulated by the ubiquitin-proteasome pathway and that Wnt signaling inhibits the process. The domain responsible for the down-regulation and direct ubiquitination was identified. We also show an unexpected role for Axin in facilitating the ubiquitination-proteasome-mediated down-regulation of APC through the oligomerization of Axin. Our results suggest a new mechanism for the regulation of APC by Axin and Wnt signaling. PMID- 15355979 TI - The FcepsilonRIbeta immunoreceptor tyrosine-based activation motif exerts inhibitory control on MAPK and IkappaB kinase phosphorylation and mast cell cytokine production. AB - The high affinity IgE Fc receptor (FcepsilonRI) beta chain functions as a signal amplifier and has been linked to atopy, asthma, and allergy. Herein, we report on a previously unrecognized negative regulatory role for the nonconventional beta chain immunoreceptor tyrosine-based activation motif that contains three tyrosine residues (YX5YX3Y). Degranulation and leukotriene production was found to be impaired in cells expressing the mutated FcepsilonRIbeta immunoreceptor tyrosine based activation motifs FYY, YYF, FYF, and FFF. In contrast, cytokine synthesis and secretion were enhanced in the YFY and FFF mutants. FcepsilonRI phosphorylation and Lyn kinase co-immunoprecipitation was intact in the YFY mutant but was lost in the FYF and FFF mutants. The phosphorylation of Syk, LAT, phospholipase gamma1/2, and Srchomology 2 domain-containing protein phosphatase 2 was intact, whereas the phosphorylation of SHIP-1 was significantly reduced in the YFY mutant cells. The FYF and FFF mutants were defective in phosphorylating all of these molecules. In contrast, the phosphorylation of ERK, p38 MAPK, IkappaB kinase beta (IKKbeta), and nuclear NFkappaB activity was enhanced in the YFY and FFF mutants. These findings show that the FcepsilonRIbeta functions to both selectively amplify (degranulation and leukotriene secretion) and dampen (lymphokine) mast cell effector responses. PMID- 15355980 TI - Presynaptic trafficking of synaptotagmin I is regulated by protein palmitoylation. AB - Protein palmitoylation plays a critical role in sorting and targeting of several proteins to pre- and postsynaptic sites. In this study, we have analyzed the role of palmitoylation in trafficking of synaptotagmin I and its modulation by synaptic activity. We found that palmitoylation of N-terminal cysteines contributed to sorting of synaptotagmin I to an intracellular vesicular compartment at the presynaptic terminal. Presynaptic targeting is a unique feature of N-terminal sequences of synaptotagmin I because the palmitoylated N terminus of synaptotagmin VII failed to localize to presynaptic sites. We also found that palmitate was stably associated with both synaptotagmin I and SNAP-25 and that rapid neuronal depolarization did not affect palmitate turnover on these proteins. However, long-term treatment with drugs that either block synaptic activity or disrupt SNARE complex assembly modulated palmitoylation and accumulation of synaptotagmin I at presynaptic sites. We conclude that palmitoylation is involved in trafficking of specific elements involved in transmitter release and that distinct mechanisms regulate addition and removal of palmitate on select neuronal proteins. PMID- 15355981 TI - Kinome profiling for studying lipopolysaccharide signal transduction in human peripheral blood mononuclear cells. AB - The DNA array technique allows comprehensive analysis of the genome and transcriptome, but the high throughput array-based assessment of intracellular signal transduction remains troublesome. The goal of this study was to test a new peptide array technology for studying the activity of all kinases of whole cell lysates, the kinome. Cell lysates from human peripheral blood mononuclear cells before and after stimulation with lipopolysaccharide were used for in vitro phosphorylation with [gamma-33P]ATP arrays consisting of 192 peptides (substrates for kinases) spotted on glass. The usefulness of peptide arrays for studying signal transduction was demonstrated by the generation of the first comprehensive description of the temporal kinetics of phosphorylation events induced by lipopolysaccharide stimulation. Furthermore analysis of the signals obtained suggested activation of p21Ras by lipopolysaccharide, and this was confirmed by direct measurement of p21Ras GTP levels in lipopolysaccharide-stimulated human peripheral blood mononuclear cells, which represents the first direct demonstration of p21Ras activation by stimulation of a Toll receptor family member. Further confidence in the usefulness of peptide array technology for studying signal transduction came from Western blot analysis of lipopolysaccharide-stimulated cells, which corroborated the signals obtained using peptide arrays as well as from the demonstration that kinase inhibitors effected peptide array phosphorylation patterns consistent with the expected action of these inhibitors. We conclude that this first metabolic array is a useful method to determine the enzymatic activities of a large group of kinases, offering high throughput analysis of cellular metabolism and signal transduction. PMID- 15355982 TI - Identification and characterization of a Pi isoform of glutathione S-transferase (GSTP1) as a zeaxanthin-binding protein in the macula of the human eye. AB - Uptake, metabolism, and stabilization of xanthophyll carotenoids in the retina are thought to be mediated by specific xanthophyll-binding proteins (XBPs). A membrane-associated XBP was purified from human macula using ion-exchange chromatography followed by gel-exclusion chromatography. Two-dimensional gel electrophoresis showed a prominent spot of 23 kDa and an isoelectric point of 5.7. Using mass spectral sequencing methods and the public NCBI database, it was identified as a Pi isoform of human glutathione S-transferase (GSTP1). Dietary (3R,3'R)-zeaxanthin displayed the highest affinity with an apparent Kd of 0.33 microm, followed by (3R,3'S-meso)-zeaxanthin with an apparent Kd of 0.52 microm. (3R,3'R,6'R)-Lutein did not display any high-affinity binding to GSTP1. Other human recombinant glutathione S-transferase (GST) proteins, GSTA1 and GSTM1, exhibited only low affinity binding of xanthophylls. (3R,3'S-meso)-Zeaxanthin, an optically inactive nondietary xanthophyll carotenoid present in the human macula, exhibited a strong induced CD spectrum in association with human macular XBP that was nearly identical to the CD spectrum induced by GSTP1. Like-wise, dietary (3R,3'R)-zeaxanthin displayed alterations in its CD spectrum in association with GSTP1 and XBP. Other mammalian xanthophyll carrier proteins such as tubulin, high density lipoprotein, low-density lipoprotein, albumin, and beta-lactoglobulin did not bind zeaxanthins with high affinity, and they failed to induce or alter xanthophyll CD spectra to any significant extent. Immunocytochemistry with an antibody to GSTP1 on human macula sections showed highest labeling in the outer and inner plexiform layers. These results indicate that GSTP1 is a specific XBP in human macula that interacts with (3R,3'S-meso)-zeaxanthin and dietary (3R,3'R) zeaxanthin in contrast to apparently weaker interactions with (3R,3'R,6'R) lutein. PMID- 15355983 TI - Secretion of sterols and the NPC2 protein from primary astrocytes. AB - Astrocytes secrete cholesterol in lipoprotein particles. Here we show that primary murine embryonic astrocytes secrete endogenously synthesized cholesterol but also the cholesterol precursors desmosterol and lathosterol. In astrocyte membranes, desmosterol and cholesterol were the predominant sterols. Astrocytes derived from Niemann-Pick type C lipidosis (NPC1-/-) mice displayed late endosomal cholesterol deposits, but the secretion of biosynthetic sterols from the cells was not inhibited. Both wild-type and NPC1-/- astrocytes secreted the NPC2 protein. Size-exclusion chromatography combined with electron microscopy showed that the majority of sterols were secreted separately from NPC2 in heterogeneous spherical particles with an average diameter of 20 nm. These data suggest that NPC2 and the majority of sterols secreted from astrocytes are not released together and that the secretion of neither sterols nor NPC2 requires NPC1 function. In addition, the findings reveal a complexity of sterol species in astrocytes and bring up the possibility that some of the effects assigned to astrocyte cholesterol may be attributed to its penultimate precursors. PMID- 15355984 TI - Construction of a cyclin D1-Cdk2 fusion protein to model the biological functions of cyclin D1-Cdk2 complexes. AB - Cyclin D1 is frequently overexpressed in human breast cancers, and cyclin D1 overexpression correlates with poor prognosis. Cyclin D1-Cdk2 complexes were previously observed in human breast cancer cell lines, but their role in cell cycle regulation and transformation was not investigated. This report demonstrates that Cdk2 in cyclin D1-Cdk2 complexes from mammary epithelial cells is phosphorylated on the activating phosphorylation site, Thr(160). Furthermore, cyclin D1-Cdk2 complexes catalyze Rb phosphorylation on multiple sites in vitro. As a model to investigate the biological and biochemical functions of cyclin D1 Cdk2 complexes, and the mechanisms by which cyclin D1 activates Cdk2, a cyclin D1 Cdk2 fusion gene was constructed. The cyclin D1-Cdk2 fusion protein expressed in epithelial cells was phosphorylated on Thr(160) and catalyzed the phosphorylation of Rb on multiple sites in vitro and in vivo. Kinase activity was not observed if either the cyclin D1 or Cdk2 domain was mutationally inactivated. Mutational inactivation of the cyclin D1 domain prevented activating phosphorylation of the Cdk2 domain on Thr(160). These results indicate that the cyclin D1 domain of the fusion protein activated the Cdk2 domain through an intramolecular mechanism. Cells stably expressing the cyclin D1-Cdk2 fusion protein exhibited several hallmarks of transformation including hyperphosphorylation of Rb, resistance to TGFbeta-induced growth arrest, and anchorage-independent proliferation in soft agar. We propose that cyclin D1-Cdk2 complexes mediate some of the transforming effects of cyclin D1 and demonstrate that the cyclin D1-Cdk2 fusion protein is a useful model to investigate the biological functions of cyclin D1-Cdk2 complexes. PMID- 15355985 TI - Bacteria binding by DMBT1/SAG/gp-340 is confined to the VEVLXXXXW motif in its scavenger receptor cysteine-rich domains. AB - The scavenger receptor cysteine-rich (SRCR) proteins form an archaic group of metazoan proteins characterized by the presence of SRCR domains. These proteins are classified in group A and B based on the number of conserved cysteine residues in their SRCR domains, i.e. six for group A and eight for group B. The protein DMBT1 (deleted in malignant brain tumors 1), which is identical to salivary agglutinin and lung gp-340, belongs to the group B SRCR proteins and is considered to be involved in tumor suppression and host defense by pathogen binding. In a previous study we used nonoverlapping synthetic peptides covering the SRCR consensus sequence to identify a 16-amino acid bacteria-binding protein loop (peptide SRCRP2; QGRVEVLYRGSWGTVC) within the SRCR domains. In this study, using overlapping peptides, we pinpointed the minimal bacteria-binding site on SRCRP2, and thus DMBT1, to an 11-amino acid motif (DMBT1 pathogen-binding site 1 or DMBT1pbs1; GRVEVLYRGSW). An alanine substitution scan revealed that VEVL and Trp are critical residues in this motif. Bacteria binding by DMBT1pbs1 was different from the bacteria binding by the macrophage receptor MARCO in which an RXR motif was critical. In addition, the homologous consensus sequences of a number of SRCR proteins were synthesized and tested for bacteria binding. Only consensus sequences of DMBT1 orthologues bound bacteria by this motif. PMID- 15355986 TI - Stable interaction between beta-arrestin 2 and angiotensin type 1A receptor is required for beta-arrestin 2-mediated activation of extracellular signal regulated kinases 1 and 2. AB - Binding of beta-arrestins to seven-membrane-spanning receptors (7MSRs) not only leads to receptor desensitization and endocytosis but also elicits additional signaling processes. We recently proposed that stimulation of the angiotensin type 1A (AT(1A)) receptor results in independent beta-arrestin 2- and G protein mediated extracellular signal-regulated kinases 1 and 2 (ERK1/2) activation. Here we utilize two AT(1A) mutant receptors to study these independent pathways, one truncated at residue 324, thus removing all potential carboxyl-terminal phosphorylation sites, and the other bearing four mutations in the serine/threonine-rich clusters in the carboxyl terminus. As assessed by confocal microscopy, the two mutant receptors interacted with beta-arrestin 2-green fluorescent protein with much lower affinity than did the wild-type receptor. In addition, the mutant receptors more robustly stimulated G protein-mediated inositol phosphate production. Approximately one-half of the wild-type AT(1A) receptor-stimulated ERK1/2 activation was via a beta-arrestin 2-dependent pathway (suppressed by beta-arrestin 2 small interfering RNA), whereas the rest was mediated by a G protein-dependent pathway (suppressed by protein kinase C inhibitor). ERK1/2 activation by the mutant receptors was insensitive to beta arrestin 2 small interfering RNA but was reduced more than 80% by a protein kinase C inhibitor. The biochemical consequences of ERK activation by the G protein and beta-arrestin 2-dependent pathways were also distinct. G-protein mediated ERK activation enhanced the transcription of early growth response 1, whereas beta-arrestin 2-dependent ERK activation did not. In addition, stimulation of the truncated AT(1A) mutant receptor caused significantly greater early growth response 1 transcription than did the wild-type receptor. These findings demonstrate how the ability of receptors to interact with beta-arrestins determines both the mechanism of ERK activation as well as the physiological consequences of this activation. PMID- 15355987 TI - Extracellular zinc triggers ERK-dependent activation of Na+/H+ exchange in colonocytes mediated by the zinc-sensing receptor. AB - Extracellular zinc promotes cell proliferation and its deficiency leads to impairment of this process, which is particularly important in epithelial cells. We have recently characterized a zinc-sensing receptor (ZnR) linking extracellular zinc to intracellular release of calcium. In the present study, we addressed the role of extracellular zinc, acting via the ZnR, in regulating the MAP kinase pathway and Na+/H+ exchange in colonocytes. We demonstrate that Ca2+ release, mediated by the ZnR, induces phosphorylation of ERK1/2, which is highly metal-specific, mediated by physiological concentrations of extracellular Zn2+ but not by Cd2+, Fe2+, Ni2+, or Mn2+. Desensitization of the ZnR by Zn2+, is followed by approximately 90% inhibition of the Zn2+ -dependent ERK1/2 phosphorylation, indicating that the ZnR is a principal link between extracellular Zn2+ and ERK1/2 activation. Application of both the IP3 pathway and PI 3-kinase antagonists largely inhibited Zn2+ -dependent ERK1/2 phosphorylation. The physiological significance of the Zn2+ -dependent activation of ERK1/2 was addressed by monitoring Na+/H+ exchanger activity in HT29 cells and in native colon epithelium. Preincubation of the cells with zinc was followed by robust activation of Na+/H+ exchange, which was eliminated by cariporide (0.5 microm); indicating that zinc enhances the activity of NHE1. Activation of NHE1 by zinc was totally blocked by the ERK1/2 inhibitor, U0126. Prolonged acidification, in contrast, stimulates NHE1 by a distinct pathway that is not affected by extracellular Zn2+ or inhibitors of the MAP kinase pathway. Desensitization of ZnR activity eliminates the Zn2+ -dependent, but not the prolonged acidification dependent activation of NHE1, indicating that Zn2+ -dependent activation of H+ extrusion is specifically mediated by the ZnR. Our results support a role for extracellular zinc, acting through the ZnR, in regulating multiple signaling pathways that affect pH homeostasis in colonocytes. Furthermore activation of both, ERK and NHE1, by extracellular zinc may provide the mechanism linking zinc to enhanced cell proliferation. PMID- 15355988 TI - p14 Arf promotes small ubiquitin-like modifier conjugation of Werners helicase. AB - Here we demonstrate a novel p53-independent interaction between the nucleolar tumor suppressors, p14 Arf and Werners helicase (WRN). Binding of p14 Arf to WRN is multivalent and resembles the binding of p14 Arf to Mdm2. Residues 2-14 and 82 101 of p14 Arf and residues in the central region and C terminus of WRN have particular importance for binding. p14 Arf promotes small ubiquitin-like modifier (SUMO) modification of WRN in a synergistic manner with the SUMO-conjugating enzyme, UBCH9. p14 Arf causes redistribution of WRN within the nucleus, and this effect is reversed by expression of a SUMO-specific protease, thus implicating the SUMO conjugation pathway in WRN re-localization. We establish that the ability to promote SUMO conjugation is a general property of the p14 Arf tumor suppressor. PMID- 15355989 TI - Novel down-regulatory mechanism of the surface expression of the vasopressin V2 receptor by an alternative splice receptor variant. AB - In rat kidney, two alternatively spliced transcripts are generated from the V2 vasopressin receptor gene. The large transcript (1.2 kb) encodes the canonical V2 receptor, whereas the small transcript encodes a splice variant displaying a distinct sequence corresponding to the putative seventh transmembrane domain and the intracellular C terminus of the V2 receptor. This work showed that the small spliced transcript is translated in the rat kidney collecting tubules. However, the protein encoded by the small transcript (here called the V2b splice variant) is retained inside the cell, in contrast to the preferential surface distribution of the V2 receptor (here called the V2a receptor). Cells expressing the V2b splice variant do not exhibit binding to 3H-labeled vasopressin. Interestingly, we found that expression of the splice variant V2b down-regulates the surface expression of the V2a receptor, most likely via the formation of V2a.V2b heterodimers as demonstrated by co-immunoprecipitation and fluorescence resonance energy transfer experiments between the V2a receptor and the V2b splice variant. The V2b splice variant would then be acting as a dominant negative. The effect of the V2b splice variant is specific, as it does not affect the surface expression of the G protein-coupled interleukin-8 receptor (CXCR1). Furthermore, the sequence encompassing residues 242-339, corresponding to the C-terminal domain of the V2b splice variant, also down-regulates the surface expression of the V2a receptor. We suggest that some forms of nephrogenic diabetes insipidus are due to overexpression of the splice variant V2b, which could retain the wild-type V2a receptor inside the cell via the formation of V2a.V2b heterodimers. PMID- 15355990 TI - Bi-directional regulation between tyrosine kinase Etk/BMX and tumor suppressor p53 in response to DNA damage. AB - Etk/Bmx, a member of the Tec family of nonreceptor tyrosine kinases, has been implicated in the regulation of various cellular processes including proliferation, differentiation, motility, and apoptosis. Here, we report the identification of Tec family kinases as the potential interacting proteins of the tumor suppressor p53 by an Src homology 3 domain array screening. Etk is physically associated with p53 through its Src homology 3 domain and the proline rich domain of p53. Induction of p53 expression by DNA damage inhibits Etk activity in several cell types. Down-regulation of Etk expression by a specific small interfering RNA sensitizes prostate cancer cells to doxorubicin-induced apoptosis, suggesting that inhibition of Etk activity is required for apoptosis in response to DNA damage. We also show that Etk primarily interacts with p53 in the cytoplasm and that such interaction leads to bidirectional inhibition of the activities of both proteins. Overexpression of Etk in prostate cancer cells results in inhibition of p53 transcriptional activity and its interaction with the mitochondrial protein BAK and confers the resistance to doxorubicin. Therefore, we propose that the stoichiometry between p53 and the Tec family kinases in a given cell type may determine its sensitivity to chemotherapeutic drugs. PMID- 15355991 TI - Structural characterization of an ATPase active F1-/V1 -ATPase (alpha3beta3EG) hybrid complex. AB - Co-reconstitution of subunits E and G of the yeast V-ATPase and the alpha and beta subunits of the F(1)-ATPase from the thermophilic Bacillus PS3 (TF(1)) resulted in an alpha(3)beta(3)EG hybrid complex showing 53% of the ATPase activity of TF(1). The alpha(3)beta(3)EG oligomer was characterized by electron microscopy. By processing 40,000 single particle projections, averaged two dimensional projections at 1.2-2.4-nm resolution were obtained showing the hybrid complex in various positions. Difference mapping of top and side views of this complex with projections of the atomic model of the alpha(3)beta(3) subcomplex from TF(1) (Shirakihara, Y., Leslie, A. G., Abrahams, J. P., Walker, J. E., Ueda, T., Sekimoto, Y., Kambara, M., Saika, K., Kagawa, Y., and Yoshida, M. (1997) Structure 5, 825-836) demonstrates that a seventh mass is located inside the shaft of the alpha(3)beta(3) barrel and extends out from the hexamer. Furthermore, difference mapping of the alpha(3)beta(3)EG oligomer with projections of the A(3)B(3)E and A(3)B(3)EC subcomplexes of the V(1) from Caloramator fervidus (Chaban, Y., Ubbink-Kok, T., Keegstra, W., Lolkema, J. S., and Boekema, E. J. (2002) EMBO Rep. 3, 982-987) shows that the mass inside the shaft is made up of subunit E, whereby subunit G was assigned to belong at least in part to the density of the protruding stalk. The formation of an active alpha(3)beta(3)EG hybrid complex indicates that the coupling subunit gamma inside the alpha(3)beta(3) oligomer of F(1) can be effectively replaced by subunit E of the V-ATPase. Our results have also demonstrated that the E and gamma subunits are structurally similar, despite the fact that their genes do not show significant homology. PMID- 15355992 TI - ErbB4 expression in neural progenitor cells (ST14A) is necessary to mediate neuregulin-1beta1-induced migration. AB - Activation of the receptor tyrosine kinase ErbB4 leads to various cellular responses such as proliferation, survival, differentiation, and chemotaxis. Two pairs of naturally occurring ErbB4 isoforms differing in their juxtamembrane (JMa/JMb) and C termini (cyt1/cyt2) have been described. To examine the role of ErbB4 in neuron migration, we cloned and stably transfected each of the four ErbB4 isoforms in ST14A cells (a neural progenitor cell line derived from the striatum of embryonic day 14 rats) endogenously expressing the other members of the ErbB family: ErbB1, ErbB2, and ErbB3. Using immunoprecipitation assays, we showed that the neuregulin-1beta1 (NRG1beta1) stimulus induced ErbB4 tyrosine phosphorylation and phosphatidylinositol 3-kinase (PI3K) recruitment and activation (as demonstrated by Akt phosphorylation) either directly (ErbB4 cyt1 isoform) or indirectly (ErbB4 cyt2 isoform). We examined the ability of the four ErbB4 isoforms to induce chemotaxis and cell proliferation in response to NRG1beta1 stimulation. Using migration assays, we observed that only ErbB4 expressing cells stimulated with NRG1beta1 showed a significant increase in migration, whereas the growth rate remained unchanged. Additional assays showed that inhibition of PI3K (but not of phospholipase Cgamma) dramatically reduced migratory activity. Our data show that ErbB4 signaling via PI3K activation plays a fundamental role in controlling NRG1beta1-induced migration. PMID- 15355993 TI - First snapshots of the HIV-1 RNA structure in infected cells and in virions. AB - With the increasing interest of RNAs in regulating a range of cell biological processes, very little is known about the structure of RNAs in tissue culture cells. We focused on the 5'-untranslated region of the human immunodeficiency virus type 1 RNA genome, a highly conserved RNA region, which contains structural domains that regulate key steps in the viral replication cycle. Up until now, structural information only came from in vitro studies. Here, we developed chemical modification assays to test nucleotide accessibility directly in infected cells and viral particles, thus circumventing possible biases and artifacts linked to in vitro assays. The secondary structure of the 5' untranslated region in infected cells points to the existence of the various stem loop motifs associated to distinct functions, proposed from in vitro probing, mutagenesis, and phylogeny. However, compared with in vitro data, subtle differences were observed in the dimerization initiation site hairpin, and none of the proposed long range interactions were observed between the functional domains. Moreover, no global RNA rearrangement was observed; structural differences between infected cells and viral particles were limited to the primer binding site, which became protected against chemical modification upon tRNA(3) (Lys) annealing in virions and to the main packaging signal. In addition, our data suggested that the genomic RNA could already dimerize in the cytoplasm of infected cells. Taken together, our results provided the first analysis of the dynamic of RNA structure of the human immunodeficiency virus type 1 RNA genome during virus assembly ex vivo. PMID- 15355994 TI - Glucocorticoid ligands specify different interactions with NF-kappaB by allosteric effects on the glucocorticoid receptor DNA binding domain. AB - Glucocorticoids inhibit inflammation by acting through the glucocorticoid receptor (GR) and powerfully repressing NF-kappaB function. Ligand binding to the C-terminal of GR promotes the nuclear translocation of the receptor and binding to NF-kappaB through the GR DNA binding domain. We sought how ligand recognition influences the interaction between NF-kappaB and GR. Both dexamethasone (agonist) and RU486 (antagonist) promote efficient nuclear translocation, and we show occupancy of the same intranuclear compartment as NF-kappaB with both ligands. However, unlike dexamethasone, RU486 had negligible activity to inhibit NF-kappaB transactivation. This failure may stem from altered co-factor recruitment or altered interaction with NF-kappaB. Using both glutathione S-transferase pull down and bioluminescence resonance energy transfer approaches, we identified a major glucocorticoid ligand effect on interaction between the GR and the p65 component of NF-kappaB, with RU486 inhibiting recruitment compared with dexamethasone. Using the bioluminescence resonance energy transfer assay, we found that RU486 efficiently recruited NCoR to the GR, unlike dexamethasone, which recruited SRC1. Therefore, RU486 promotes differential protein recruitment to both the C-terminal and DNA binding domain of the receptor. Importantly, using chromatin immunoprecipitation, we show that impaired interaction between GR and p65 with RU486 leads to reduced recruitment of the GR to the NF-kappaB-responsive region of the interleukin-8 promoter, again in contrast to dexamethasone that significantly increased GR binding. We demonstrate that ligand-induced conformation of the GR C-terminal has profound effects on the functional surface generated by the DNA binding domain of the GR. This has implications for understanding ligand-dependent interdomain communication. PMID- 15355995 TI - Camptothecin-induced imbalance in intracellular cation homeostasis regulates programmed cell death in unicellular hemoflagellate Leishmania donovani. AB - Leishmania, a unicellular trypanosomatid protozoan parasite, causes a wide range of human diseases ranging from the localized self-healing cutaneous lesions to fatal visceral leishmaniasis. However, it undergoes a process of programmed cell death during treatment with the topoisomerase I poison camptothecin (CPT). The present study shows that CPT-induced formation of reactive oxygen species increases the level of cytosolic calcium through the release of calcium ions from intracellular stores as well as by influx of extracellular calcium. Elevation of cytosolic calcium is responsible for depolarization of mitochondrial membrane potential (DeltaPsim), which is followed by a significant decrease in intracellular pH levels. CPT-induced oxidative stress also causes impairment of the Na+ - K+ -ATPase pump and subsequently decreases the intracellular K+ level in leishmanial cells. A decrease in both intracellular pH and K+ levels propagates the apoptotic process through activation of caspase 3-like proteases by rapid formation of cytochrome c-mediated apoptotic complex. In addition to caspase-like protease activation, a lower level of intracellular K+ also enhances the activation of apoptotic nucleases at the late stage of apoptosis. This suggests that the physiological level of pH and K+ are inhibitory for apoptotic DNA fragmentation and caspase-like protease activation in leishmanial cells. Moreover, unlike mammalian cells, the intracellular ATP level gradually decreases with an increase in the number of apoptotic cells after the loss of DeltaPsim. Taken together, the elucidation of biochemical events, which tightly regulate the process of growth arrest and death of Leishmania donovani promastigotes, allows us to define a more comprehensive view of cell death during treatment with CPT. PMID- 15355996 TI - Targeted knockdown of the RNA-binding protein CRD-BP promotes cell proliferation via an insulin-like growth factor II-dependent pathway in human K562 leukemia cells. AB - The c-myc mRNA coding region determinant-binding protein (CRD-BP) was first identified as a masking protein that stabilizes c-myc mRNA in a cell-free mRNA degradation system. Thus, CRD-BP is thought to promote cell proliferation by maintaining c-Myc at critical levels. CRD-BP also appears to be an oncofetal protein, based upon its expression during mammalian development and in some tumors. By using K562 leukemia cells as a model, we show that CRD-BP gene silencing by RNA interference significantly promoted proliferation, indicating an inhibitory effect of CRD-BP on proliferation. Unexpectedly, CRD-BP knockdown had no discernible effect on c-myc mRNA levels. CRD-BP is also known as insulin-like growth factor II (IGF-II) mRNA-binding protein-1. It has been reported to repress translation of a luciferase reporter mRNA containing an IGF-II 5'-untranslated region known as leader 3 but not one containing IGF-II leader 4. CRD-BP knockdown markedly increased IGF-II mRNA and protein levels but did not alter translation of luciferase reporter mRNAs containing 5'-untranslated regions consisting of either IGF-II leader 3 or leader 4. Addition of antibody against IGF-II to cell cultures inhibited the proliferative effect of CRD-BP knockdown, suggesting that regulation of IGF-II gene expression, rather than c-myc mRNA levels, mediates the proliferative effect of CRD-BP knockdown. Thus, we have identified a dominant function for CRD-BP in cell proliferation of human K562 cells, involving a possible IGF-II-dependent mechanism that appears independent of its ability to serve as a c-myc mRNA masking protein. PMID- 15355997 TI - Tuberin binds p27 and negatively regulates its interaction with the SCF component Skp2. AB - TSC1 (tuberous sclerosis complex 1) encoding hamartin and TSC2 encoding tuberin are tumor suppressor genes responsible for the autosomal dominantly inherited disease tuberous sclerosis. These genes have been demonstrated to negatively regulate cell cycle progression, the activity of cdk2, and the degradation of the cyclin-dependent kinase inhibitor p27. To date, the underlying molecular mechanism remains elusive. Here, we show that tuberin binds to p27. Whereas tuberin also binds p27 in TSC1-negative cells, hamartin does not bind p27 without tuberin. p27 protein levels are regulated through ubiquitin-dependent degradation. Skp2 is the F-box protein, which, together with other proteins, forms an SCF (Skp1/cullin/F-box protein)-type E3 ubiquitin ligase complex whose task is to target p27 for degradation by the proteasome. We found that neither tuberin nor hamartin are in a complex with Skp2. Tuberin does not affect Skp2 protein levels, and the SCFSkp2 ubiquitin ligase does not regulate tuberin stability. But binding of tuberin to p27 sequesters p27 from Skp2 accompanied by an up-regulation of the p27 interaction with cdk2. Skp2-induced p27 degradation and cell cycle progression is abolished by tuberin's protective binding to p27. This work, the first description of the direct interaction of a tumor suppressor protein with p27, provides a molecular explanation for the effects of tuberous sclerosis complex genes on the cell cycle and demonstrates a new aspect of the SCFSkp2-mediated regulation of p27 stability. PMID- 15355998 TI - A yeast mitochondrial membrane methyltransferase-like protein can compensate for oxa1 mutations. AB - Members of the Oxa1p/Alb3/YidC family mediate the insertion of various organelle or bacterial hydrophobic proteins into membranes. They present at least five transmembrane segments (TM) linked by hydrophilic domains located on both sides of the membrane. To examine how Oxa1p structure relates to its function, we have introduced point mutations and large deletions into various domains of the yeast mitochondrial protein. These mutants allowed us to show the importance of the first TM domain as well as a synergistic interaction between the first loop and the C-terminal tail, which both protrude into the matrix. These mutants also led to the isolation of a high copy suppressor, OMS1, which encodes a member of the methyltransferase family. Overexpression of OMS1 seems to increase the steady state level of both the mutant and wild-type Oxa1p. We show that Oms1p is a mitochondrial inner membrane protein inserted independently of Oxa1p. Oms1p presents one TM and a N-in C-out topology with the C-terminal domain carrying the methyltransferase-like domain. A conserved motif within this domain is essential for the suppression of oxa1 mutations. We discuss the possible role of Oms1p on Oxa1p intermembrane space domain. PMID- 15355999 TI - A lipid peroxidation-derived inflammatory mediator: identification of 4-hydroxy-2 nonenal as a potential inducer of cyclooxygenase-2 in macrophages. AB - Cyclooxygenases (COXs) catalyze the conversion of arachidonic acid to eicosanoids, which mediate a variety of biological actions involved in vascular pathophysiology. In the present study, we investigated the role of lipid peroxidation products in the up-regulation of COX-2, an inducible isoform responsible for high levels of prostaglandin production during inflammation and immune responses. COX-2 was found to colocalize with 4-hydroxy-2-nonenal (HNE), a major lipid peroxidation-derived aldehyde, in foamy macrophages within human atheromatous lesions, suggesting that COX-2 expression may be associated with the accumulation of lipid peroxidation products within macrophages. To test the hypothesis that lipid peroxidation products might be involved in the regulation of prostanoid biosynthesis, we conducted a screen of oxidized fatty acid metabolites and found that, among the compounds tested, only HNE showed inducibility of the COX-2 protein in RAW264.7 macrophages. In addition, intraperitoneal administration of HNE resulted in an increase in cell numbers in the peritoneal cavity that was associated with significant increases in the peritoneal and tissue levels of COX-2 in mice. To understand the possible signaling mechanism underlying the inducing effect of HNE on COX-2 up-regulation, we examined the phosphorylation events that may lead to COX-2 induction and found that HNE did not stimulate the induction of nitric oxide synthase and activation of NF-kappaB but significantly activated p38 mitogen-activated protein kinase and its upstream kinase in RAW264.7 macrophages. Tyrosine kinases, such as the epidermal growth factor-like and Src family tyrosine kinases, appeared to mediate the stabilization of COX-2 mRNA via the p38 mitogen-activated protein kinase pathway. These findings suggest that HNE accumulated in macrophages/foam cells may represent an inflammatory mediator that plays a role in stimulation of the inflammatory response and contributes to the progression of atherogenesis. PMID- 15356000 TI - (S)-2,3-Di-O-geranylgeranylglyceryl phosphate synthase from the thermoacidophilic archaeon Sulfolobus solfataricus. Molecular cloning and characterization of a membrane-intrinsic prenyltransferase involved in the biosynthesis of archaeal ether-linked membrane lipids. AB - The core structure of membrane lipids of archaea have some unique properties that permit archaea to be distinguished from the others, i.e. bacteria and eukaryotes. (S)-2,3-Di-O-geranylgeranylglyceryl phosphate synthase, which catalyzes the transfer of a geranylgeranyl group from geranylgeranyl diphosphate to (S)-3-O geranylgeranylglyceryl phosphate, is involved in the biosynthesis of archaeal membrane lipids. Enzymes of the UbiA prenyltransferase family are known to catalyze the transfer of a prenyl group to various acceptors with hydrophobic ring structures in the biosynthesis of respiratory quinones, hemes, chlorophylls, vitamin E, and shikonin. The thermoacidophilic archaeon Sulfolobus solfataricus was found to encode three homologues of UbiA prenyltransferase in its genome. One of the homologues encoded by SSO0583 was expressed in Escherichia coli, purified, and characterized. Radio-assay and mass spectrometry analysis data indicated that the enzyme specifically catalyzes the biosynthesis of (S)-2,3-di-O geranylgeranylglyceryl phosphate. The fact that the orthologues of the enzyme are encoded in almost all archaeal genomes clearly indicates the importance of their functions. A phylogenetic tree constructed using the amino acid sequences of some typical members of the UbiA prenyltransferase family and their homologues from S. solfataricus suggests that the two other S. solfataricus homologues, excluding the (S)-2,3-di-O-geranylgeranylglyceryl phosphate synthase, are involved in the production of respiratory quinone and heme, respectively. We propose here that archaeal prenyltransferases involved in membrane lipid biosynthesis might be prototypes of the protein family and that archaea might have played an important role in the molecular evolution of prenyltransferases. PMID- 15356001 TI - The Soh1/MED31 protein is an ancient component of Schizosaccharomyces pombe and Saccharomyces cerevisiae Mediator. AB - We here demonstrated that the Soh1/MED31 protein is a stable component of Mediator complex isolated from Schizosaccharomyces pombe and Saccharomyces cerevisiae. Bioinformatic analysis traces the Soh1/MED31 family of Mediator subunits to the point of major eukaryotic divergence, before the appearance of the canonical heptapeptide repeat structure of the RNA polymerase II C-terminal domain. PMID- 15356002 TI - Structural, thermodynamic, and kinetic analyses of tetrahydrooxazine-derived inhibitors bound to beta-glucosidases. AB - The understanding of transition state mimicry in glycoside hydrolysis is increasingly important both in the quest for novel specific therapeutic agents and for the deduction of enzyme function and mechanism. To aid comprehension, inhibitors can be characterized through kinetic, thermodynamic, and structural dissection to build an "inhibition profile." Here we dissect the binding of a tetrahydrooxazine inhibitor and its derivatives, which display Ki values around 500 nm. X-ray structures with both a beta-glucosidase, at 2 A resolution, and an endoglucanase at atomic (approximately 1 A) resolution reveal similar interactions between the tetrahydrooxazine inhibitor and both enzymes. Kinetic analyses reveal the pH dependence of kcat/Km and 1/Ki with both enzyme systems, and isothermal titration calorimetry unveils the enthalpic and entropic contributions to beta-glucosidase inhibition. The pH dependence of enzyme activity mirrored that of 1/Ki in both enzymes, unlike the cases of isofagomine and 1-deoxynojirimycin that have been characterized previously. Calorimetric dissection reveals a large favorable enthalpy that is partially offset by an unfavorable entropy upon binding. In terms of the similar profile for the pH dependence of 1/Ki and the pH dependence of kcat/Km, the significant enthalpy of binding when compared with other glycosidase inhibitors, and the tight binding at the optimal pH of the enzymes tested, tetrahydrooxazine and its derivatives are a significantly better class of glycosidase inhibitor than previously assumed. PMID- 15356003 TI - Calcium-dependent self-assembly of human centrin 2. AB - Human centrin 2 (HsCen2) is a member of the EF-hand superfamily of calcium binding proteins, often associated with the centrosomes and basal bodies. These organelles exhibit different morphological aspects, including a variety of centrin-containing fibers that connect the two centrioles or other structural elements of the pericentriolar space. The molecular basis of the Ca(2+)-sensitive fibers and their precise role in centrosome duplication are not known. To explore the possible structural role of HsCen2, we initiated a physicochemical study of the self-assembly properties of the purified protein in vitro. Using light scattering experiments, we investigated the temporal evolution of the assembly process and characterized the dependence on various chemical and physical factors, including temperature, di-cation concentration, ionic strength, protein concentration, and pH. The reversible self-assembly revealed many features of a large-size protein polymerization, with nucleation and elongation steps. Kinetic and equilibrium experiments show that a hydrophobic fluorescent probe (ANS) inhibits the polymerization by interfering with the nucleation step, probably through interactions with the apolar exposed sites on the protein surface. A truncated form of HsCen2, lacking the first 25 residues (Delta25HsCen2), shows no detectable self-assembly, pointing to the critical role played by the N-terminal fragment in the supermolecular organization of HsCen2. As revealed by isothermal titration experiments, the isolated N-terminal domains bind with a significant affinity (2 x 10(5) m(-1)) to preformed oligomers of Delta25HsCen2 through an entropy-driven mechanism. PMID- 15356004 TI - Src tyrosine kinase inhibitor PP2 markedly enhances Ras-independent activation of Raf-1 protein kinase by phorbol myristate acetate and H2O2. AB - Recently we reported that simultaneous treatment of NIH 3T3 cells with the combination of phorbol myristate acetate (PMA) and hydrogen peroxide (H2O2) resulted in synergistic activation of Raf-1 kinase (Lee, M., Petrovics, G., and Anderson, W. B. (2003) Biochem. Biophys. Res. Commun. 311, 1026-1033). In this study we have demonstrated that PP2 (4-amino-5-(4-chlorophenyl)-7-(t butyl)pyrazolo[3,4-d]pyrimidine), a potent and selective inhibitor of the Src family tyrosine kinase, greatly potentiated the ability of PMA and/or H2O2 to activate Raf-1 kinase, whereas it blocked the tyrosine phosphorylation of Raf-1. Unlike PMA/H2O2 treatment, which showed transient activation, PP2-mediated Raf-1 activation was sustained and continued to increase through 4 h of treatment. Transient transfection studies with a dominant-negative mutant of Ras (N19Ras) indicated that this PP2-induced activation of Raf-1 was Ras-independent. Moreover, PP2 showed no effect on platelet-derived growth factor-induced Raf-1 activation. Interestingly, mutation of the reported Raf-1 Src family tyrosine kinase phosphorylation site by conversion of tyrosines 340 and 341 to phenylalanine (YY340/341FF Raf) had limited effect on the ability of PP2 to induce significant stimulation of Raf-1 kinase activity. Taken together, our results suggest that a tyrosine phosphorylation event is involved in the negative feedback regulation of Raf-1. Inhibition of a Src family tyrosine kinase by PP2 appears to alleviate this tyrosine kinase-mediated inhibition of Raf-1 and allow activating modification(s) of Raf-1 to proceed. This PP2 effect resulted in significant and sustained Ras-independent activation of Raf-1 by PMA and H2O2. PMID- 15356005 TI - Evidence for and characterization of Ca2+ binding to the catalytic region of Arabidopsis thaliana phospholipase Dbeta. AB - Most types of plant phospholipase D (PLD) require Ca(2+) for activity, but how Ca(2+) affects PLD activity is not well understood. We reported previously that Ca(2+) binds to the regulatory C2 domain that occurs in the N terminus of the Ca(2+)-requiring PLDs. Using Arabidopsis thaliana PLDbeta and C2-deleted PLDbeta (PLDbetacat), we now show that Ca(2+) also interacts with the catalytic regions of PLD. PLDbetacat exhibited Ca(2+)-dependent activity, was much less active, and required a higher level of Ca(2+) than the full-length PLDbeta. Ca(2+) binding of the proteins was stimulated by phospholipids; phosphatidylserine was the most effective among those tested. Scatchard plot analysis of Ca(2+) binding data yielded an estimate of 3.6 high affinity (K(d) = 29 mum) binding sites on PLDbeta. The Ca(2+)-PLDbetacat interaction increased the affinity of the protein for the activator, phosphatidylinositol 4,5-bisphosphate, but not for the substrate, phosphatidylcholine. This is in contrast to the effect of Ca(2+) binding to the C2 domain, which stimulates phosphatidylcholine binding but inhibits phosphatidylinositol 4,5-bisphosphate binding of the domain. These results demonstrate the contrasting and complementary effects of the Ca(2+)- and lipid-binding properties of the C2 and catalytic domains of plant PLD and provide insight into the mechanism by which Ca(2+) regulates PLD activity. PMID- 15356006 TI - Diet, insulin resistance, and obesity: zoning in on data for Atkins dieters living in South Beach. AB - Insulin resistance is a central pathogenic factor for the metabolic syndrome and is associated with both generalized obesity and the accumulation of fat in the omental and intramyocellular compartments. In the context of the current obesity epidemic, it is imperative to consider diets in terms of their ability to both promote weight loss and ameliorate insulin resistance. Weight loss under any dietary formulation depends on hypocaloric intake, and only moderate weight loss (5-10%) is sufficient to augment insulin sensitivity. However, increments in insulin sensitivity may be more directly related to loss of intramyocellular or omental fat rather than loss of total body weight per se. The widespread acceptance of popular low-carbohydrate high-fat diets (e.g. Atkins Diet, Zone Diet, South Beach diet) further underscores the need to evaluate dietary interventions regarding their safety and metabolic effects. These high-fat diets have been shown to be safe in the short term; however, their long-term safety has not been established. With respect to insulin sensitivity, diets enriched in saturated fats can induce insulin resistance, whereas fat substitution with monounsaturated fats can enhance insulin sensitivity. On the other hand, high fiber, high-carbohydrate diets comprised of foods with low caloric density can similarly be used for effective weight reduction and to ameliorate insulin resistance. Although some data suggest that low-glycemic index diets are most advantageous in this regard, these effects may have more to do with increments in dietary fiber than differences in available carbohydrates. Popular low carbohydrate, high-fat diets are being fervently embraced as an alternative to challenging modifications in lifestyle and intentional calorie reduction. Current data do not support such unbridled enthusiasm for these diets, particularly in relationship to high-fiber, high-carbohydrate diets emphasizing intake of fresh vegetables and fruits. Long-term studies to determine the efficacy and safety of both popular and experimental diets are warranted. PMID- 15356007 TI - Regional adiposity and insulin resistance. PMID- 15356008 TI - Childhood obesity. PMID- 15356009 TI - Aldosterone-renin ratios in the context of primary care. PMID- 15356010 TI - Aldosterone to Renin ratio in a primary care setting: the Bussolengo study. AB - The aim of the study was to evaluate the prevalence of hypertension associated with an elevated aldosterone to renin activity ratio (ARR) in a sample of adults aged 35-74 yr, randomly selected from the population register of the Bussolengo Health District (northern Italy) and representative of the total population of the district. Subjects (n = 1462) were randomly selected from the population register and examined by their general practitioners. Complete data for 1348 individuals were available for final statistical analysis. Apart from verapamil or alpha-blockers, no hypotensive drugs were allowed during the 4 wk before assay. Direct active renin and aldosterone were measured in the plasma of hypertensive patients after 2 h in the upright posture. Of 412 identified hypertensive patients, 287 subjects agreed to give blood (70% response rate). An aldosterone to active renin ratio (AARR) of 32 pg/ml was taken as the cut-off value, equivalent to an ARR of 50 ng/dl/ng/ml.h. An elevated AARR was observed in 32.4% of the hypertensive patients, with increased prevalence in females and in people aged 55 yr or older. As an elevated AARR is frequent in the general hypertensive population, screening should not be limited to the patients referred to specialist units. PMID- 15356011 TI - Radioactive iodine use in childhood Graves' disease: time to wake up and smell the I-131. PMID- 15356012 TI - A 36-year retrospective analysis of the efficacy and safety of radioactive iodine in treating young Graves' patients. AB - This report details the 26- and 36-yr outcomes of 116 patients under the age of 20 yr with Graves' disease who were treated with radioiodine between 1953 and 1973. Contacted by telephone and mail in 1991-1992, 107 of them supplied personal historical data, and their physicians furnished interval histories, physical examinations, and laboratory data. This was repeated in 2001-2002, with 98 of them being contacted. At the time of treatment, the patients' ages ranged between 3 yr, 7 months and 19 yr, 9 months. Six were less than 6 yr of age, 11 were between 6 and 11 yr, 45 were between 11 and 15 yr, and 45 were between 16 and 19 yr. The average length of follow-up in 1991-1992 was 26.1 yr; that in 2001-2002 was 36.2 yr. None of the patients developed cancer of the thyroid or leukemia. Early on, when the objective of treatment was euthyroidism, the dose of radioiodine was low, and retreatment was frequently needed. Later, the doses used were increased. Over time, all but two patients became hypothyroid. Pregnancies did not result in an unusual number of congenital anomalies or spontaneous abortions. Treating young people with Graves' disease with radioiodine is safe and effective over the long term. PMID- 15356013 TI - Phenotypic consequences of the peroxisome proliferator-activated receptor-gamma Pro12Ala polymorphism: the weight of the evidence in genetic association studies. PMID- 15356014 TI - Ala12Ala genotype of the peroxisome proliferator-activated receptor gamma2 protects against atherosclerosis. AB - A mutation in the peroxisome proliferator-activated receptor gamma2 (PPARgamma2) gene with a cytosine to guanine substitution results in an exchange of proline (Pro) with alanine (Ala) in exon B (codon 12) of this gene. This polymorphism has been associated with high insulin sensitivity and low body weight, but no data have been published to date about its effect on early atherosclerosis. We investigated the relationship of the Pro(12)Ala polymorphism to early atherosclerosis, measured by the intima-media thickness (IMT). A total of 622 subjects were included, aged 40-70 yr, who were participants of the RIAD (Risk factors in Impaired glucose tolerance for Atherosclerosis and Diabetes) study and were at risk of developing type 2 diabetes. Altogether, 449 of the subjects had the common genotype (Pro(12)Pro), 162 had the Pro(12)Ala genotype, and 11 the Ala(12)Ala genotype. IMT was significantly decreased in subjects with the Ala(12)Ala genotype compared with subjects with the other two genotypes. Body mass index, free fatty acid levels, and leukocyte count were lower in subjects with the Ala(12)Ala genotype compared with subjects with the Pro(12)Pro or Pro(12)Ala genotypes. In multivariate analysis, the Ala(12)Ala genotype was a significant independent determinant of IMT. Furthermore, we demonstrated specific expression of the PPARgamma2 gene in human atherosclerotic lesions as well as in cultured primary macrophages and foam cells. In conclusion, our data suggest that the Ala(12)Ala genotype of the PPARgamma2 gene may protect from early atherosclerosis in subjects at risk for diabetes. PMID- 15356015 TI - Vascular calcification and osteoporosis--the nature of the nexus. PMID- 15356016 TI - Aortic calcification and the risk of osteoporosis and fractures. AB - We investigated the relation between computed tomography measures of aortic calcification and values for bone density and the number of fragility fractures in 2348 healthy, postmenopausal women. To determine whether increases in vascular calcification and bone loss progress in parallel, baseline values were compared with measurements obtained 9 months to 8 yr later in a subgroup of 228 women. Of the 2348 subjects studied, 70% had osteoporosis, 30% had at least one vertebral fracture, and 9% had at least one hip fracture. Aortic calcifications were inversely related to bone density and directly related to fractures. After adjusting for age and potential confounders, measures for aortic calcification predicted 26.1% of the variance in bone density (P < 0.001). Compared with women without calcification, the odds ratios for vertebral and hip fractures in those with calcification were estimated to be 4.8 (95% confidence interval, 3.6-6.5) and 2.9 (95% confidence interval, 1.8-4.8), respectively. The subgroup analysis of 228 women longitudinally studied showed that the percentage of yearly increase in aortic calcification accounted for 47% of the variance in the percentage rate of bone loss (P < 0.001). Moreover, a strong graded association was observed between the progression of vascular calcification and bone loss for each quartile. Women in the highest quartile for gains in aortic calcification had four times greater yearly bone loss (5.3 vs.1.3% yearly; P < 0.001) than women of similar age in the lowest quartile. Smaller, but highly significant differences were also found between all other quartiles. We conclude that aortic calcifications are a strong predictor for low bone density and fragility fractures. PMID- 15356017 TI - Leptin and appetite control in lipodystrophy. PMID- 15356018 TI - Effects of exogenous leptin on satiety and satiation in patients with lipodystrophy and leptin insufficiency. AB - To examine leptin's role in human appetite regulation, we studied recombinant methionyl human leptin's effects on satiation and satiety in a model of leptin insufficiency, lipodystrophy. Eight females with hypoleptinemia and lipodystrophy were given sc injections of A-100 (maximal dose, 200% of that predicted to normalize serum leptin) for 4 months. Satiation and satiety were determined before and again during leptin treatment. Satiation was measured as the time to voluntary cessation of eating from a standardized food array after a 12-h fast. Satiety was determined as the time to hunger sufficient to consume a full meal after consumption of a standardized preload. During leptin treatment, satiation time decreased (41.2 +/- 18.2 to 19.5 +/- 10.6 min; P = 0.01), satiety time increased (62.9 +/- 64.8 to 137.8 +/- 91.6 min; P = 0.04), energy consumed to produce satiation decreased (2034 +/- 405 to 1135 +/- 432 kcal or 8.5 +/- 1.7 to 4.7 +/- 1.8 MJ; P < 0.01), and the amount of food desired in the postabsorptive state decreased (P < 0.02). Ghrelin concentrations also decreased during leptin administration (284.3 +/- 127.9 to 140.6 +/- 104.5 pmol/liter; P < 0.002). We conclude that increased leptin in patients with lipodystrophy results in less caloric, shorter, more satiating meals and longer-lived satiety. These data support the hypothesis that leptin plays an important, permissive role in human appetite regulation. PMID- 15356019 TI - Challenging dogma in thyroid cancer molecular genetics--role of RET/PTC and BRAF in tumor initiation. PMID- 15356020 TI - BRAF mutations are not a major event in post-Chernobyl childhood thyroid carcinomas. AB - The BRAF gene has been shown to be a major target for mutations in papillary thyroid carcinoma (PTC) (36-69%), which forms almost all of the over 2000 cases of thyroid carcinoma that have occurred in Chernobyl. BRAF is activated by point mutation, and were it to occur at a high frequency in Chernobyl-related tumors, it would challenge the dominant role of double-strand breaks in radiation-induced PTC. In a previous study, we detected the BRAF V600E mutation in 46% (23 of 50) of sporadic adult PTC. Using the same methodology, we have analyzed 34 post Chernobyl PTC and detected RET/PTC rearrangements in 14 (41%) and BRAF mutations (V600E) in four (12%). These two alterations did not coexist in any PTCs. The mean age at exposure of patients with PTC showing BRAF mutation was higher than that of patients with tumors without BRAF mutation irrespective of their RET status. We have also analyzed 17 sporadic cases of childhood PTC and found that only one (6%) harbored the BRAF V600E mutation. We conclude that the frequency of BRAF mutations is significantly lower (P = 0.0008) in post-Chernobyl PTC than in adult sporadic PTC, whereas no significant difference was found between post Chernobyl and sporadic childhood PTCs. PMID- 15356021 TI - Heterogeneity in the distribution of RET/PTC rearrangements within individual post-Chernobyl papillary thyroid carcinomas. AB - The nuclear disaster that occurred in Chernobyl in 1986 offered the unique opportunity to study the molecular genetics of one human tumor type, papillary carcinoma of the thyroid gland, associated with a specific etiology. We have analyzed RET rearrangements in post-Chernobyl papillary thyroid carcinomas (n = 29), follicular thyroid adenomas (n = 2), and follicular thyroid carcinoma (n = 1) by interphase fluorescence in situ hybridization (FISH) analysis on paraffin embedded tissue sections. Paraffin sections were microdissected before use to ensure that only tumor was present. Cell nuclei were scored for the presence of a split FISH signal (separated red and green signal) in addition to an overlapping signal. Only cells with either two overlapping signals or one split and one overlapping signal were counted to ensure that only complete cell nuclei had been scored. In total, 23 of 32 cases (72%) showed RET rearrangements diagnosed by FISH interphase analysis. In all cases, the tumors were composed of a mixture of cells with and without ret rearrangement on FISH. In some cases, this distribution was clearly nonrandom because clustering of rearranged cells was detected within the same tumor nodule. Accordingly, only 31% of the cases positive for rearrangement on FISH also scored positive using RT-PCR. These findings suggest that because RET/PTC rearrangements are not present in a majority of tumor cells, either a fraction of post-Chernobyl papillary thyroid tumors are of multiclonal origin, or ret rearrangement is a later, subclonal event. PMID- 15356022 TI - Low frequency of BRAFT1796A mutations in childhood thyroid carcinomas. AB - A high prevalence of the activating BRAF mutation, BRAF(T1796A), is observed in adult papillary thyroid carcinomas (PTCs). The prognosis of childhood PTCs is generally fairly good despite the fact that distant metastases are often documented in these cases. To investigate the differences between the characteristics of childhood and adult PTCs, we analyzed both BRAF(T1796A) and RAS mutations in 31 Japanese and 48 post-Chernobyl Ukrainian thyroid carcinomas. In the 31 Japanese childhood cases, BRAF(T1796A) was found in only one instance (3.2%), and no RAS mutations were detected. In the Ukrainian subjects, of the 15 childhood and the 33 adolescent and young adult PTCs examined, the BRAF(T1796A) mutation was found in zero and eight cases, respectively, and RAS mutations were found in two of the young adult cases. In addition, 17 of the 48 Ukrainian cases showed expression of the RET tyrosine kinase region, indicating the existence of RET/PTC rearrangements. Unlike adult PTCs, we could detect no positive association between BRAF(T1796A) mutations and clinical parameters in the childhood carcinomas, suggesting that a low prevalence of BRAF(T1796A) is a common feature of PTCs in children regardless of radiation exposure levels. The differences in the prevalence of BRAF(T1796A) mutations between childhood and adult cases of PTC may well reflect inherent differences in the clinical features of these cancers between the two age groups. PMID- 15356023 TI - Characterization of a novel loss of function mutation of PAX8 in a familial case of congenital hypothyroidism with in-place, normal-sized thyroid. AB - Thyroid dysgenesis is the most common cause of congenital hypothyroidism, a relatively frequent disease affecting 1 in 3000-4000 newborns. Whereas most cases are sporadic, mutations in transcription factors implicated in thyroid development have been shown to cause a minority of cases transmitted as monogenic Mendelian diseases. PAX8 is one of these transcription factors, and so far, five mutations have been identified in its paired domain in patients with thyroid dysgenesis. We have identified a novel mutation of PAX8, in the heterozygous state, in a father and his two children both presenting with congenital hypothyroidism associated with an in-place thyroid of normal size at birth. In addition, one of the affected siblings displayed unilateral kidney agenesis. The mutation substitutes a highly conserved serine in position 54 of the DNA-binding domain of the protein (S54G mutation) by a glycine. Functional analyses of the mutant protein (PAX8-S54G) demonstrated that it is unable to bind a specific cis element of the thyroperoxidase gene promoter in EMSAs and that it has almost completely lost the ability to act in synergy with Titf1 to transactivate transcription from the thyroglobulin promoter/enhancer. These results indicate that loss of function mutations of the PAX8 gene may cause congenital hypothyroidism in the absence of thyroid hypoplasia. PMID- 15356024 TI - The clinical significance of the POLG gene polymorphism in male infertility. AB - Based on association studies, an increasing number of gene polymorphisms have been proposed as modulators of spermatogenesis. Interestingly, a clear cause effect relationship between a polymorphism of the POLG gene and oligo(astheno)zoospermia was recently described. The POLG gene contains a polymorphic CAG repeat, and the presence of a homozygous mutant (not10/not10 CAG) genotype was found only in infertile men. In the present study, a large number of infertile patients and normospermic men of Italian origin were studied to define the effect of POLG genotypes on spermatogenic potential and whether the homozygous mutant is specific for spermatogenic disturbances. The mutated genotype was found at the same frequency in both infertile and normospermic men. Mean values of sperm parameters such as sperm count, motility, and morphology did not differ significantly between carriers of the three different genotypes. Our study failed to confirm any influence of the POLG gene polymorphism on the efficiency of the spermatogenesis. More importantly, considering that the homozygous mutant genotype has been found in normospermic fertile men, the analysis of the CAG repeat tract of the POLG gene does not appear to have any clinical diagnostic value. PMID- 15356025 TI - The skeletal phenotype of men with previous constitutional delay of puberty. AB - It is presently unclear whether men with a history of constitutional delay (CD) of puberty are osteopenic. This study compares auxology, bone mass, size, and density of 32 men (age, 21-33 yr) with previous CD with 45 controls. Using dual energy x-ray absorptiometry, areal bone mineral density (aBMD) and volumetric bone mineral density at the lumbar spine (LS) and femoral neck (FN), hip strength analysis, and total-body and body segment (arms, legs, trunk) measurements were determined. Auxological variables, body composition, the muscle-bone relation, and the effect of prior androgen treatment were studied. Men with previous CD were shorter (P < 0.001) and had shorter height-adjusted arms compared with controls. Height-adjusted total-body bone mineral content (BMC) (P = 0.004), aBMD (P = 0.016), and bone area (P = 0.006) but not lean tissue mass (P = 0.507) were lower in CD men compared with controls; consequently, their BMC to lean tissue mass ratio was reduced (P < 0.001). Segment length-adjusted BMC and bone area of arms (P < 0.001) and legs (P < 0.03), but not trunk were lower in CD men than in controls. They had lower LS aBMD (P = 0.044) but normal LS and FN volumetric bone mineral density. Size-adjusted LS width and the hip cross-sectional area were lower than in controls. There was no difference in anthropometric or dual-energy x-ray absorptiometry results between untreated (n = 15) and androgen-treated (n = 17) CD men.We conclude that men with previous CD have normal LS and FN volumetric density but reduced total-body bone mass, which was explained by reduced limb bone mass and size. Together with the reduced LS bone width and hip crosssectional area, these skeletal characteristics suggest impaired periosteal expansion during puberty. The skeletal phenotype of CD males may be altered by their late onset of puberty. PMID- 15356026 TI - Effect of pioglitazone on circulating adipocytokine levels and insulin sensitivity in type 2 diabetic patients. AB - We examined the effect of pioglitazone (PIO) on circulating adipocytokine levels to elucidate the mechanisms by which thiazolidinediones improve insulin resistance in type 2 diabetes mellitus (T2DM). Twenty-three subjects with T2DM (age 54 +/- 2 yr, body mass index 29 +/- 1 kg/m(2)) were randomly assigned to receive placebo (n = 11) or PIO, 45 mg/d (n = 12), for 4 months. Before and after treatment, subjects received a 75-g oral glucose tolerance test (OGTT); euglycemic insulin clamp (40 mU/m(2).min) with 3-(3)H-glucose; determination of fat mass ((3)H(2)O); and measurement of fasting glucose, free fatty acids (FFAs), leptin, adiponectin, and TNFalpha concentrations. After 4 months of PIO, fasting plasma glucose concentration (Delta = -2.7 mol/liter), mean plasma glucose during OGTT (Delta = -3.8 mol/liter), and hemoglobin A(1c) (Delta = 1.7%) decreased (P < 0.05 vs. placebo) without change in fasting or post-OGTT plasma insulin levels. Fasting FFAs (Delta = 168 micromol/liter) and TNFalpha (Delta = 0.7 pg/ml) concentrations decreased (P < 0.05 vs. placebo), whereas adiponectin (Delta = 8.7 microg/ml) increased (P < 0.01 vs. placebo). Despite the increase in body fat mass (Delta = 3.4 kg) after PIO, plasma leptin concentration did not change significantly. No changes in plasma glucose, FFAs, or adipocytokine levels were observed in placebo-treated subjects. During the insulin clamp, endogenous (hepatic) glucose production decreased (Delta = -2.67 micromol/fat-free mass.min, P < 0.05 vs. placebo), whereas metabolic clearance rate of glucose (MCR) increased (Delta = 0.58 ml/fat-free mass.min, P < 0.05 vs. placebo) after PIO. In all subjects, before and after PIO, the decrease in plasma FFA concentration was correlated with the changes in both endogenous (hepatic) glucose production (r = 0.47, P < 0.05) and MCR (r = -0.41, P < 0.05), whereas the increase in plasma adiponectin concentration was correlated with the change in endogenous (hepatic) glucose production (r = -0.70, P < 0.01) and MCR (r = 0.49, P < 0.05). These results suggest that the direct effects of PIO on adipose tissue to decrease plasma FFA levels and increase plasma adiponectin contribute to the improvements in hepatic and peripheral insulin sensitivity and glucose tolerance in patients with T2DM. PMID- 15356027 TI - Pituitary magnetic resonance imaging is not required in the postoperative follow up of acromegalic patients with long-term biochemical cure after transsphenoidal surgery. AB - After successful transsphenoidal surgery for acromegaly, life-long follow-up is required, because 10-15% of patients develop recurrence of disease. We assessed whether it is safe to perform postoperative follow-up with only biochemical evaluation in acromegalic patients initially cured by transsphenoidal surgery. We studied 32 patients cured after transsphenoidal surgery for acromegaly during a follow-up of 8.7 +/- 6.4 yr (mean +/- sd). Serial measurements of serum GH during glucose tolerance test and magnetic resonance imaging (MRI) scans were performed. Serial MRI scans were reevaluated by three independent neuroradiologists, who were blinded for the clinical and biochemical data, for growth of suspected tumor tissue. Twenty-three patients remained biochemically cured in the long term, whereas nine of the 32 patients developed recurrence of disease, indicated by elevated serum GH concentrations during glucose tolerance test and clinical symptoms/signs. None of the 23 patients with long-term biochemical cure showed growth of tumor tissue according to two neuroradiologists, whereas the third neuroradiologist assessed three of 23 patients as having tumor growth despite continuing biochemical cure. In the nine patients with biochemical recurrence, no tumor growth was found in the series of postoperative MRI scans, according to two of the three independent radiologists, whereas the third radiologist found tumor growth in four patients with recurrent disease. In conclusion, in patients with acromegaly, initially cured by transsphenoidal surgery, it appears safe to check for recurrent disease during long-term follow-up of these patients by biochemical markers only. PMID- 15356028 TI - Osteoporosis, mineral metabolism, and serum soluble tumor necrosis factor receptor p55 in viral cirrhosis. AB - Liver cirrhosis is a risk factor for osteoporosis. Nevertheless, little is known about the mechanisms of bone mass loss in patients with viral cirrhosis. TNFalpha is a potent bone-resorbing agent. Serum concentrations of soluble TNF receptor p55 (sTNFR-55) correlate with clinical activity in liver cirrhosis. Our aim was to evaluate the possible role of sTNFR-55 in the pathogenesis of osteoporosis in patients with viral cirrhosis and its relationship with bone turnover markers. We studied 40 consecutive patients with viral cirrhosis and no history of alcohol intake and 26 healthy volunteers. Bone mineral density (BMD) was measured by dual x-ray absorptiometry in the lumbar spine (LS) and femoral neck (FN). Patients with viral cirrhosis had reduced BMD (expressed as the z-score) in all sites [LS, -1.5 +/- 0.22 (P < 0.001); FN, -0.37 +/- 0.15 (P < 0.01)]. Serum concentrations of sTNFR-55 and urinary deoxypyridinoline, a biochemical marker of bone resorption, were significantly higher in patients with osteoporosis than in patients without osteoporosis (P < 0.001 and P < 0.05, respectively). Serum levels of sTNFR-55 correlated inversely with BMD in LS (r = -0.62; P < 0.005) and FN (r = -0.47; P < 0.05) and positively with urinary deoxypyridinoline (r = 0.72, P < 0.001). Our findings show that high serum concentrations of sTNFR-55 play a role in the pathogenesis of viral cirrhosis-associated bone mass loss and provide evidence of increased bone resorption related to the high serum sTNFR-55 levels. PMID- 15356029 TI - Insulin sensitization for girls with precocious pubarche and with risk for polycystic ovary syndrome: effects of prepubertal initiation and postpubertal discontinuation of metformin treatment. AB - Among girls with precocious pubarche (PP), those with low birth weight (LBW) are, even if nonobese, at risk for progression to polycystic ovary syndrome (PCOS) including hyperinsulinemic hyperandrogenism, dyslipidemia, dysadipocytokinemia, and central fat excess. Recently, we disclosed the efficacy of insulin sensitization with metformin to disrupt progression from PP to PCOS in formerly LBW girls who were postmenarche. In LBW-PP girls, we have now extended the exploration of early insulin sensitization therapy in two directions: 1) metformin therapy was started before puberty; and 2) we assessed the effects of metformin discontinuation in girls who had started metformin treatment after menarche. Prepubertal LBW-PP girls (n = 33; mean age, 8.0 yr; body mass index, 18.5 kg/m(2)) were randomly assigned to remain untreated or to receive metformin (425 mg/d) for 6 months. Postpubertal LBW-PP girls (n = 24; age, 12.4 yr; body mass index, 21.0 kg/m(2)) had been randomized (at -12 months) to remain untreated or to receive metformin (850 mg/d) for 12 months, at which time (0 month) a treatment cross-over was performed for 6 months. Fasting blood glucose and serum insulin, SHBG, dehydroepiandrosterone sulfate, androstenedione, testosterone, lipid profile, IL-6, and adiponectin were assessed at 0 and 6 months, as was body composition (by dual x-ray absorptiometry). In the prepubertal study (group A), comparisons of untreated vs. treated girls disclosed normalizing effects of metformin on SHBG, androstenedione, dehydroepiandrosterone sulfate, low and high density lipoprotein cholesterol, triglycerides, IL-6, adiponectin, total and abdominal fat mass, and lean body mass. In the postpubertal study (group B), treatment cross-over at 0 month was in each subgroup followed by a striking reversal in the course of the endocrine-metabolic state, adipocytokinemia, and body composition; all changes pointed to normalizing effects of metformin treatment. In conclusion, these two studies provide the first evidence that 1) prepubertal metformin therapy has normalizing effects on PCOS features in high risk girls with a combined history of LBW and PP; and 2) in adolescence, metformin's normalizing effects are reversed as soon as metformin therapy is discontinued. PMID- 15356030 TI - A single luteinizing hormone determination 2 hours after depot leuprolide is useful for therapy monitoring of gonadotropin-dependent precocious puberty in girls. AB - Long-acting GnRH analogs represent the standard treatment for gonadotropin dependent precocious puberty. The aim of this study was to determine the hormonal parameters for monitoring the adequacy of depot leuprolide acetate treatment in girls with clinical and hormonal diagnosis of gonadotropin-dependent precocious puberty. Eighteen girls were treated monthly with 3.75 mg depot leuprolide acetate. Adequate hypothalamic-pituitary-gonadal axis suppression during treatment was achieved in 16 of the 18 girls according to the clinical parameters and prepubertal LH levels. In these 16 well-controlled girls, the LH peak after a classical GnRH test was compared with a single LH measurement obtained 2 h after depot leuprolide acetate administration before and during GnRH analog treatment. Before therapy, the mean +/- sd LH peak after a classical GnRH test was 18.4 +/- 11.2 IU/liter (ranging from 7-41.5 IU/liter), and it was 22.6 +/- 8.3 IU/liter 2 h after the first depot leuprolide dose (ranging from 10-35.3 IU/liter). During therapy, the mean +/- sd of LH peak after classical GnRH test was 1.4 +/- 0.6 IU/liter (ranging from <0.6 to 2.3 IU/liter), and it was 2.7 +/- 1.9 IU/liter (ranging from 0.7-6.6 IU/liter) 2 h after depot leuprolide. The LH peak after a classical GnRH test and that 2 h after depot leuprolide administration correlate significantly before and during treatment. In conclusion, we established the LH cut-off values for an adequate depot leuprolide therapy as an LH peak below 2.3 IU/liter after a classical GnRH test or below 6.6 IU/liter 2 h after depot leuprolide. The latter measurement may replace the classical GnRH test as a reliable and convenient tool for monitoring therapy in female gonadotropin dependent precocious puberty. PMID- 15356031 TI - Polycystic ovarian morphology with regular ovulatory cycles: insights into the pathophysiology of polycystic ovarian syndrome. AB - To determine the relevance of polycystic ovarian morphology (PCOM) to the pathophysiology of polycystic ovarian syndrome (PCOS), biochemical features associated with PCOS were examined in 68 women with an established history of regular ovulatory cycles and no clinical evidence of hyperandrogenism. Ovarian morphology was objectively assessed by pelvic ultrasound. LH, FSH, estradiol (E(2)), testosterone (T), androstenedione (Delta(4)A), SHBG, and dehydroepiandrosterone sulfate (DHEAS) were measured at baseline in the early follicular phase (EFP) in all subjects. LH, FSH, E(2), and progesterone (P(4)) were then measured daily for a complete menstrual cycle in 16 women with normal ovarian morphology and in 26 women with PCOM. T, Delta(4)A, SHBG, and DHEAS levels were measured in pools of three daily samples in each of the EFP, midcycle, and midluteal phases. An additional 26 normal women (13 with normal ovarian morphology and 13 with PCOM) were studied in the EFP to assess pulsatile LH secretion, insulin and glucose levels, and the ovarian response to human chorionic gonadotropin. At baseline, there were no differences in body mass index or hirsutism scores between women with PCOM and normal ovaries. In daily samples across the menstrual cycle LH, FSH, E(2), and P(4) did not differ between women with PCOM and those with normal ovaries, and there was no difference in LH pulse amplitude or frequency in the EFP frequent sampling studies. In women with PCOM, T (P < 0.01), free T (P < 0.005), and DHEAS (P < 0.01) levels were higher at baseline in the EFP, and SHBG was lower (P < 0.05). Differences in Delta(4)A did not reach significance (P = 0.14). T, free T, Delta(4)A, and DHEAS were also increased in PCOM across the menstrual cycle (P < 0.05). In addition, 17 hydroxyprogesterone (P < 0.02), Delta(4)A (P < 0.01), and T (P < 0.01) responses to human chorionic gonadotropin were greater in women with PCOM. Fasting glucose was not different between the two groups, but fasting insulin was higher (P < 0.02) in PCOM women as was insulin resistance calculated from homeostatic model assessment (P < 0.01). These studies demonstrate that PCOM in nonhirsute women with documented ovulatory cycles is associated with normal E(2), P(4), and gonadotropin dynamics, but higher androgen and insulin levels and lower SHBG levels. Taken together, these findings suggest that PCOM with ovulatory cycles exists as a discrete entity, represents the mildest form of ovarian hyperandrogenism, and is associated with greater insulin resistance than in women with normal ovarian morphology. The absence of any neuroendocrine abnormality in women with PCOM and ovulatory cycles suggests that gonadotropin dysfunction is not required for increased androgen secretion, but may be critical for development of the anovulatory disorder associated with PCOS. PMID- 15356032 TI - Essential amino acid and carbohydrate supplementation ameliorates muscle protein loss in humans during 28 days bedrest. AB - We determined whether essential amino acid and carbohydrate supplementation could offset the catabolic response to prolonged inactivity. Major outcome measures included mixed muscle fractional synthetic rate (FSR), phenylalanine net balance, lean leg mass, and leg extension strength. On d 1 and 28, vastus lateralis muscle biopsies and femoral arterio-venous blood samples were obtained during a primed constant infusion of l-[ring-(2)H(5)]phenylalanine. Net balance and FSR were calculated over 16 h, during which the control group (CON) received a nutritionally mixed meal every 5 h (0830, 1330, and 1830 h). The experimental group (EXP) also consumed 16.5 g essential amino acids and 30 g carbohydrate (1100, 1600, and 2100 h). The dietary regimen was maintained during bedrest. FSR was higher in the EXP group on d 1 (EXP, 0.099 +/- 0.008%/h; CON: 0.075 +/- 0.005%/h) and d 28 (EXP, 0.093 +/- 0.006%/h; CON, 0.055 +/- 0.007%/h). Lean leg mass was maintained throughout bedrest in the EXP group (+0.2 +/- 0.3 kg), but fell in the CON group (-0.4 +/- 0.1 kg). Strength loss was more pronounced in the CON group (EXP, -8.8 +/- 1.4 kg; CON, -17.8 +/- 4.4 kg). Essential amino acid and carbohydrate supplementation may represent a viable intervention for individuals at risk of sarcopenia due to immobility or prolonged bedrest. PMID- 15356033 TI - Anemia in patients with type 1 diabetes. AB - Anemia is a common finding in diabetes, although most patients in these studies have type 2 disease. This study examines the prevalence and predictors of anemia in outpatients with type 1 diabetes. A full blood count was obtained in addition to routine testing in patients with type 1 diabetes at the Austin Medical Centre (n = 135), Melbourne, the Royal Prince Alfred Hospital (n = 42), and the Royal North Shore Hospital (n = 135), both in Sydney, Australia. One in seven patients had anemia (14%). Patients at greatest risk could be identified by the presence of diabetic kidney disease. More than half (52%) of patients with macroalbuminuria had anemia, compared with 24% of patients with microalbuminuria and less than 8% of normoalbuminuric patients. Patients with diabetes and renal impairment were more than six times more likely to have anemia than those with normal renal function. Patients with anemia were more likely to have retinopathy and macrovascular complications than were patients with a normal hemoglobin level, independent of comorbid renal disease. Anemia is a prevalent finding in patients with type 1 diabetes and represents a significant unrecognized burden. Patients at greatest risk can be identified by the presence of renal disease, in the form of albuminuria and/or renal impairment. PMID- 15356034 TI - Insulin resistance and proliferative retinopathy: a cross-sectional, case-control study in 115 patients with type 2 diabetes. AB - In this cross-sectional, case-control study we explored the association of proliferative diabetic retinopathy (PDR) with insulin resistance (IR) in type 2 diabetics with serum creatinine less than 2.0 mg/dl. For each PDR case, one reference case with background diabetic retinopathy (BDR) and two controls without retinopathy were identified. IR was evaluated by hyperinsulinemic euglycemic clamp; retinopathy was evaluated by indirect ophthalmoscopy and photography. Patients were matched by age, gender, and body mass index. PDR patients (n = 28) had higher IR and low-density lipoprotein cholesterol and triglyceride levels than BDR patients (n = 29), but comparable levels of glycosylated hemoglobin. Compared with patients without retinopathy (n = 58), those with PDR had higher IR, low-density lipoprotein cholesterol, and albuminuria (P < 0.05); those with BDR had higher glycosylated hemoglobin (P < 0.05), but comparable IR. At multivariate regression analysis, IR was the only independent marker of PDR among patients with retinopathy (P = 0.016). IR also retained its independent predictive value at multiple comparison among all groups (by Kruskal-Wallis test, P = 0.019). In type 2 diabetes, IR is an independent specific marker of proliferative retinopathy that may characterize patients at increased risk for blindness who may benefit most from early screening and therapeutic intervention. Longitudinal studies are needed to evaluate the role of IR in the pathogenesis of proliferative retinopathy. PMID- 15356035 TI - Increase in 6-hydroxymelatonin excretion in humans during ascent to high altitudes. AB - Melatonin (MLT), the pineal gland hormone involved in the regulation of circadian rhythms, shows characteristic diurnal variation. Its physiological role in humans is not clear. Exposure to high altitudes may disrupt the circadian rhythm and lead to various endocrine changes. MLT in humans has not been studied under these conditions. Urinary 6-hydroxy-MLT sulfate (aMT6s) excretion was analyzed during the day (0700-2200 h) and night (2200-0700 h) phases. A cohort of 33 healthy volunteers, aged 19-65 yr, was studied during an ascent to a high altitude in the Himalayas on three occasions (at a lower altitude, at 3400 m, and after reaching maximal altitudes of 5600-6100 m). aMT6s excretion during the daytime remained unchanged during exposure to high altitudes. As expected, nocturnal values were higher than diurnal values at each point in time. However, there was a significant increase in nocturnal MLT excretion after the ascent to high altitudes. Ascent to high altitudes is associated with increased nocturnal excretion of aMT6s. The mechanism and physiological significance of this MLT increase are unclear. PMID- 15356036 TI - Circulating free insulin-like growth factor (IGF)-I, total IGF-I, and IGF binding protein-3 levels do not predict the future risk to develop prostate cancer: results of a case-control study involving 201 patients within a population-based screening with a 4-year interval. AB - Recent studies have reported that serum IGF-I levels in the highest quartile of the normal range and IGF binding protein-3 (IGFBP-3) in the lowest quartile of the normal range are associated with an increased risk of future prostate cancer and/or presence of prostate cancer. It has also been suggested that the measurement of circulating total IGF-I concentrations might be a useful tool for the early detection of prostate cancer in men with moderately increased prostate specific antigen (PSA) levels. To determine whether circulating free IGF-I, total IGF-I, and IGFBP-3 levels can predict future prostate cancer risk, we prospectively studied prostate cancer characteristics in a cohort of men during two rounds (mean interval, 4 yr) of a population-based screening study for prostate cancer. Two hundred one prostate cancer cases were detected at the second-round screening (aged 55-70 yr), and all these subjects were enrolled in the case group for the present study. Prostate cancer had been confirmed by biopsy in all cases. These 201 subjects were matched with the 201 nonprostate cancer cases by age, serum PSA range at the first-round screening (PSA < 2 ng/ml, n = 67; PSA = 2-3 ng/ml, n = 67; and PSA = 3-4 ng/ml, n = 67), and residence area. At baseline, total IGF-I, free IGF-I, and IGFBP-3 levels and prostate volume of cases with prostate cancer were not different from those of healthy controls. PSA velocity was significantly different between cases and controls (P < 0.001).Stepwise forward logistic regression analysis showed that only PSA levels at baseline and PSA at round 2 after 4 yr are good predictors of prostate cancer, whereas total IGF-I, free IGF-I, and IGFBP-3 did not predict the development of prostate cancer. Only one of the 201 subjects with prostate cancer had metastases. Within the subjects with prostate cancer, there were no differences of IGF-I parameters with different tumor node metastasis categories and/or Gleason scores. Our study suggests that the measurement of serum IGF-I and/or IGFBP-3 concentrations in addition to PSA does not improve the identification of men at high risk to develop early stages of prostate cancer. In addition, our results indicate that the endocrine IGF-I system is not directly involved in the growth of the early stages of prostate cancer. PMID- 15356037 TI - Reproductive function during summer and winter in Norwegian men living north and south of the Arctic circle. AB - Seasonal, daylight-dependent variation in human spermatozoa counts, with lowest values during summer, has been suggested. To test this hypothesis, we performed a longitudinal study of semen quality and reproductive hormone levels in Norwegian men living north and south of the Arctic Circle. An ejaculate and a serum specimen were obtained both in summer and in winter from 92 volunteers in Tromsoe (69 degrees north latitude) and 112 in Oslo (60 degrees north latitude). Semen analyses were performed, and serum was assayed for FSH and inhibin B. The median spermatozoa concentration in Tromsoe after adjustment for abstinence period length was 49 x 10(6)/ml in summer and 54 x 10(6)/ml in winter. Corresponding values for Oslo were 59 x 10(6)/ml and 54 x 10(6)/ml. The seasonal differences in spermatozoa concentration were not statistically significant, nor were significant differences observed in median total spermatozoa count, semen volume, percentage progressive motile spermatozoa, or FSH. In Tromsoe, but not Oslo, inhibin B concentration was slightly, but significantly (P = 0.02) higher in winter than summer (229 ng/liter vs. 223 ng/liter). The length of the daylight period may have a slight impact on hormonal markers of spermatogenesis but does not cause substantial changes in spermatozoa numbers and motility. PMID- 15356038 TI - SHOX haploinsufficiency and Leri-Weill dyschondrosteosis: prevalence and growth failure in relation to mutation, sex, and degree of wrist deformity. AB - SHOX mutations causing haploinsufficiency were reported in Leri-Weill dyschondrosteosis (LWD), which is characterized by mesomelic short stature and Madelung deformity of the wrists. The aim of this study was to determine the prevalence of SHOX mutations in LWD and to investigate the degree of growth failure in relation to mutation, sex, age of menarche, and wrist deformity. We studied 20 families with 24 affected children (18 females) and nine affected parents (seven females). All patients presented with bilateral Madelung deformity and shortening of the limbs. Height, sitting height, parental height, birth length, age of menarche, and presence of minor abnormalities were recorded. The degree of Madelung deformity was estimated by analysis of left hand radiographs. Microsatellite typing of the SHOX locus was used for detection of SHOX deletions and PCR direct sequencing for the detection of SHOX point mutations. In 14 of 20 families (70%), SHOX mutations were detected, with seven deletions (four de novo) and seven point mutations (one de novo). The latter included five missense mutations of the SHOX homeodomain, one nonsense mutation (E102X) truncating the whole homeodomain, and one point mutation (X293R) causing a C-terminal elongation of SHOX. Median age of the affected children was 13.4 yr (range, 6.1-18.3), mean height sd score (SDS) (sd in parentheses) was -2.85 (1.04), and mean sitting height/height ratio SDS was +3.06 (1.09). Mean birth length SDS was -0.59 (1.26). Growth failure occurred before school age. Height change during a median follow up of 7.4 yr (range, 2.3-11.3) was insignificant with a mean change in height SDS of -0.10 (0.52). Mean height SDS of affected parents was -2.70 (0.85) vs. -0.91 (1.10) in unaffected parents. Height loss due to LWD was estimated calculating delta height defined by actual height SDS minus target height SDS of the unaffected parent(s). In the children, mean delta height SDS was -2.16 (1.06), the loss being greater in girls at -2.30 (1.02) than in boys at -1.72 (1.09) (P = 0.32). In patients with SHOX deletions, it was -2.14 (1.15) vs. -1.67 (0.73) for the SHOX point mutation group (P = 0.38). Mean delta height SDS was -2.26 (0.68) for the girls with early menarche (<12 yr) vs. -2.08 (0.91) for the other postmenarcheal girls (P = 0.72). Height loss in patients with radiologically severe wrist deformities in comparison with those having milder radiological signs was -2.81 (1.01) vs. -1.70 (1.04) (P = 0.03). GH treatment in five children during a median duration of 3.4 yr (range, 1.5-9.8 yr) with a median dosage of 0.23 mg/kg.wk (range, 0.14-0.25) resulted in a mean height SDS gain of +0.82 (0.34). In conclusion, SHOX defects were the main cause of LWD. Growth failure occurred during the first years of life with a mean height loss of 2.16 SDS whereas pubertal growth may only be mildly or not affected. Children with a severe degree of wrist deformity were significantly shorter than those with mild deformities. No statistically significant effects of type of mutation, age of menarche, or sex on height were observed. The effect of GH therapy varied between individuals and needs to be examined in controlled studies. PMID- 15356039 TI - Marked decrease in sleepiness in patients with sleep apnea by etanercept, a tumor necrosis factor-alpha antagonist. AB - The proinflammatory cytokines, TNFalpha and IL-6, are elevated in obstructive sleep apnea (OSA) and have been proposed as mediators of excessive daytime sleepiness in humans. We tested the effects of etanercept, a medication that neutralizes TNFalpha and is approved by the FDA for the treatment of rheumatoid arthritis, in eight obese male apneics. These patients participated in a pilot, placebo-controlled, double-blind study during which nighttime polysomnography, multiple sleep latency test, and fasting blood glucose and plasma levels of IL-6, C-reactive protein, insulin, and adiponectin were obtained. There was a significant and marked decrease in sleepiness by etanercept, which increased sleep latency during the multiple sleep latency test by 3.1 +/- 1.0 min (P < 0.05) compared with placebo. Also, the number of apneas/hypopneas per hour was reduced significantly by the drug compared with placebo (52.8 +/- 9.1 vs. 44.3 +/ 10.3; adjusted difference, -8.4 +/- 2.3; P < 0.05). Furthermore, IL-6 levels were significantly decreased after etanercept administration compared with placebo (3.8 +/- 0.9 vs. 1.9 +/- 0.4 pg/ml; adjusted difference, -1.9 +/- 0.5; P < 0.01). However, no differences were observed in etanercept vs. placebo in the levels of fasting blood glucose and plasma C-reactive protein, insulin, and adiponectin. We conclude that neutralizing TNFalpha activity is associated with a significant reduction of objective sleepiness in obese patients with OSA. This effect, which is about 3-fold higher than the reported effects of continuous positive airway pressure on objective sleepiness in patients with OSA (0.9 vs. 3.1 min), suggests that proinflammatory cytokines contribute to the pathogenesis of OSA/sleepiness. PMID- 15356040 TI - Overexpression of 11beta-hydroxysteroid dehydrogenase-1 in adipose tissue is associated with acquired obesity and features of insulin resistance: studies in young adult monozygotic twins. AB - 11beta-Hydroxysteroid dehydrogenase type 1 (11beta-HSD-1) catalyzes the interconversion of inactive cortisone to active cortisol. Overexpression of 11beta-HSD-1 in murine adipose tissue results in glucocorticoid receptor (GR)alpha overexpression, central obesity, and insulin resistance. It is controversial whether 11beta-HSD-1 or GRalpha expression are increased in human adipose tissue in obesity. We studied effects of acquired obesity on 11beta-HSD-1 gene (real-time PCR) and protein (Western blotting) expression in sc adipose tissue in 17 monozygotic twin pairs aged 24-27 yr with a mean intrapair difference in body mass index (BMI) of 3.8 kg/m(2) (range 0.4-10.1 kg/m(2)). Intrapair correlations were calculated to study effects of acquired obesity on 11beta-HSD-1 expression. Western blot analysis of adipose tissue homogenates identified approximately 50- and approximately 68-kDa proteins specific for 11beta-HSD-1. Both structural forms correlated positively with 11beta-HSD-1 mRNA concentrations. Intrapair differences in 11beta-HSD-1 mRNA, and the 50- and 68 kDa proteins in sc adipose tissue correlated positively with those in BMI (kilograms per square meter) (r = 0.78 for 11beta-HSD-1 mRNA, P = 0.0002; r = 0.87 for the 11beta-HSD-1 50-kDa protein, P = 0.0003; and r = 0.62 for the 11beta HSD-1 68-kDa protein, P = 0.033), total body fat (percent) (r = 0.65, P = 0.005; r = 0.83, P = 0.001; and r = 0.69, P = 0.013, respectively) and sc fat (cubed centimeters) (r = 0.66, P = 0.004; r = 0.94, P = 0.0001; and r = 0.71, P = 0.009, respectively). Furthermore, 11beta-HSD-1 mRNA and 50-kDa protein expression, but not 68-kDa protein expression, correlated positively with intrapair differences in intraabdominal fat mass (cubed centimeters) (r = 0.62, P = 0.008; r = 0.69, P = 0.013; r = 0.48, P = 0.112) and serum fasting insulin concentration (milliunits per liter) (r = 0.76, P = 0.0004; r = 0.60, P = 0.037; and r = 0.43, P = 0.160, respectively). Intrapair differences in GRalpha expression were significantly inversely correlated with those in BMI and total and sc fat mass. In conclusion, expression of 11beta-HSD-1 in sc adipose tissue is increased in human acquired obesity and is closely related to accumulation of sc and intraabdominal fat and features of insulin resistance. PMID- 15356041 TI - Factors that affect final height and change in height standard deviation scores in survivors of childhood cancer treated with growth hormone: a report from the childhood cancer survivor study. AB - GH deficiency is a common late complication in survivors of pediatric malignancies, particularly those who are treated with radiation (RT) to the hypothalamic-pituitary region. Nonetheless, few reports have assessed final height outcomes in survivors treated with GH. In the present study, we investigated which patient and treatment variables correlate with final height and change in height sd score (SDS) in a large cohort of cancer survivors treated with GH. We previously identified 361 participants in the multicenter Childhood Cancer Survivor Study who were treated with GH. Final height data were available in 183 survivors (120 males). Diagnoses included: central nervous system tumors (n = 90), acute leukemia (n = 64), soft tissue sarcomas (n = 23), and miscellaneous (n = 6). The median age at diagnosis of the primary cancer was 4.6 yr, and the median age at start of GH treatment was 11.3 yr. Mean height SDS at start of GH therapy was -2.03 +/- 0.8, and the mean final height SDS was -1.48 +/ 0.10 (P < 0.001). Final height SDS was positively associated with target height and dose of GH but negatively associated with the presence of concomitant endocrinopathies and dose of spinal RT. Change in height SDS (start of GH-final height) was positively associated with male gender, younger bone age at start of GH, and dose of GH; presence of concomitant endocrinopathies and dose of spinal RT were negatively associated with change in height SDS. Risk factors associated with a final height of -2.0 sd or less included lower doses of GH and exposure to higher doses of spinal RT. Thus, to maximize final height, our findings emphasize the importance of beginning GH therapy at the earliest bone age that is clinically feasible; treating with conventional higher doses of GH; and, when possible, minimizing the dose of spinal RT. PMID- 15356042 TI - Safety and efficacy of anastrozole for the treatment of pubertal gynecomastia: a randomized, double-blind, placebo-controlled trial. AB - Pubertal gynecomastia is thought to result from transient imbalances between estrogen and androgen concentrations. Anastrozole (ARIMIDEX), a potent and selective aromatase inhibitor, decreases estrogen and increases testosterone concentrations in pubertal boys. The safety and efficacy of anastrozole for the treatment of pubertal gynecomastia were evaluated. In a randomized, double-blind, placebo-controlled study of 80 boys, aged 11-18 yr, with pubertal gynecomastia that had not reduced over a 3-month interval, subjects received either anastrozole (1 mg) or placebo once daily for 6 months. A response was defined as a 50% or greater reduction in the calculated volume of both breasts combined using ultrasonography measurements. A comparison of response rates was performed using logistic regression analysis. Secondary end points included changes in serum hormone concentrations. The percentage of patients with a response was 38.5% for the anastrozole group and 31.4% for the placebo group (odds ratio, 1.513; 95% confidence interval, 0.496-4.844; P = 0.47). At 6 months, the median percent change in the testosterone/estradiol ratio was 166% for the anastrozole group and 39% for the placebo group. Anastrozole treatment was well tolerated. In patients with pubertal gynecomastia, no significant difference in the percentage of patients with a 50% or greater reduction in total breast volume, as calculated from ultrasonography measurements, was demonstrated between the anastrozole and placebo groups. PMID- 15356043 TI - Preservation of neuroendocrine control of reproductive function despite severe undernutrition. AB - Anorexia nervosa (AN) is characterized by low weight and self-imposed caloric restriction and leads to severe bone loss. Although amenorrhea due to acquired GnRH deficiency is nearly universal in AN, a subset of patients maintains menses despite low weight. The mechanisms underlying continued GnRH secretion despite low weight in these patients and the impact of gonadal hormone secretion on bone mineral density (BMD) in such eumenorrheic, low-weight patients remain unknown. We hypothesized that 1) eumenorrheic women with AN would have higher body fat and levels of nutritionally dependent hormones, including leptin and IGF-I, than amenorrheic women with AN and comparable body mass index; and 2) BMD would be higher in these women. We also investigated whether the severity of eating disorder symptomatology differed between the groups. We studied 116 women: 1) 42 low-weight women who fulfilled all Diagnostic and Statistical Manual of Mental Disorders (fourth edition) diagnostic criteria for AN, except for amenorrhea; and 2) 74 women with AN and amenorrhea for at least 3 months. The two groups were similar in body mass index (17.1 +/- 0.2 vs. 16.8 +/- 0.2 kg/m(2)), percent ideal body weight (78.2 +/- 0.8% vs. 76.7 +/- 0.8%), duration of eating disorder (70 +/ 13 vs. 59 +/- 9 months), age of menarche (13.2 +/- 0.3 vs. 13.5 +/- 0.2 yr), and exercise (4.5 +/- 1.0 vs. 4.2 +/- 0.5 h/wk). As expected, eumenorrheic patients had a higher mean estradiol level (186.6 +/- 19.0 vs. 59.4 +/- 2.5 nmol/liter; P < 0.0001) than amenorrheic subjects. Mean percent body fat, total body fat mass, and truncal fat were higher in eumenorrheic than amenorrheic patients [20.9 +/- 0.9% vs. 16.7 +/- 0.6% (P = 0.0001); 9.8 +/- 0.5 vs. 7.8 +/- 0.3 kg (P = 0.0009); 3.4 +/- 0.2 vs. 2.7 +/- 0.1 kg (P = 0.006)]. The mean leptin level was higher in the eumenorrheic compared with the amenorrheic group (3.7 +/- 0.3 vs. 2.8 +/- 0.2 ng/ml; P = 0.04). Serum IGF-I levels were also higher in the eumenorrheic than in the amenorrheic group (41.8 +/- 3.7 vs. 30.8 +/- 2.3 nmol/liter; P = 0.02). There were only minor differences in severity of eating disorder symptomatology, as measured by the Eating Disorders Inventory, and where differences were observed, eumenorrheic subjects manifested more severe symptomatology than amenorrheic subjects. Mean BMD at the posterior-anterior and lateral spine were low in both groups, but were higher in patients with eumenorrhea than in those with amenorrhea [posterior-anterior spine T-score, -0.9 +/- 0.1 vs. -1.9 +/- 0.1 (P < 0.0001); lateral spine T-score, -1.2 +/- 0.1 vs. -2.3 +/- 0.2 (P < 0.0001)]. In contrast, preservation of menstrual function was not protective at the total hip (total hip T-score, -0.9 +/- 0.1 vs. -1.1 +/- 0.1; P = 0.27), trochanter, or femoral neck. In summary, patients with eumenorrhea had more body fat and higher serum leptin levels than their amenorrheic counterparts of similar weight. Moreover, reduced bone density was observed in both groups, but was less severe at the spine, but not the hip, in women with undernutrition and preserved menstrual function than in amenorrheic women of similar weight. Therefore, fat mass may be important for preservation of normal menstrual function in severely undernourished women, and this may be in part mediated through leptin secretion. In addition, nutritional intake and normal hormonal function may be independent contributors to maintenance of trabecular bone mass in low-weight women. PMID- 15356044 TI - Propylthiouracil before 131I therapy of hyperthyroid diseases: effect on cure rate evaluated by a randomized clinical trial. AB - A randomized clinical trial was performed to clarify whether pretreatment with propylthiouracil (PTU) before radioiodine ((131)I) therapy influences the final outcome of this therapy, as has been indicated by retrospective studies. Untreated consecutive hyperthyroid patients with Graves' disease (n = 23) or a toxic nodular goiter (n = 57) were randomized to either PTU (+PTU; n = 39) or no pretreatment (-PTU; n = 41) before compensated (131)I therapy. The median PTU dose was 100 mg, which was discontinued 4 d before treatment. The median (131)I activity was 302 MBq (range, 87-600 MBq). After (131)I therapy, the serum free T(4) index increased in the +PTU group from 97.7 +/- 47.5(+/-sd) nmol/liter at the time of therapy to 152.3 +/- 77.6 nmol/liter at 3 wk (P < 0.001) and 140.4 +/ 75.9 nmol/liter at 6 wk (P < 0.001). In the -PTU group, the serum free T(4) index, which was initially 254.3 +/- 145.7 nmol/liter, decreased significantly to 212.0 +/- 113.0 nmol/liter at 3 wk (P < 0.05) and 165.8 +/- 110.0 nmol/liter at 6 wk (P < 0.005). After 1 yr of follow-up, the treatment failure rate in patients with a toxic nodular goiter was four times higher in the +PTU group than in the PTU group (nine of 20 vs. three of 25 patients; P = 0.06), whereas the difference among patients with Graves' disease was less obvious (four of six vs. four of nine; P = 0.81). Patients in the +PTU group who were cured had higher serum TSH (s-TSH) levels at the time of (131)I therapy than those who were not cured. By adjusting for a possible interfactorial relationship through a regression analysis, including the s-TSH level and type of disease, only PTU pretreatment had a significant adverse effect on the cure rate (P = 0.03). In conclusion, this randomized trial demonstrates that PTU pretreatment reduces the cure rate of (131)I therapy in hyperthyroid diseases, although this adverse effect seems to be attenuated by the concomitant rise in s-TSH. PMID- 15356045 TI - Prolactin release is enhanced in proportion to excess visceral fat in obese women. AB - Prolactin (PRL) promotes (visceral) fat accrual in a variety of animal models. The release of PRL by the pituitary is tonically inhibited by dopamine through activation of the dopamine D2 receptor (D2R) of lactotroph cells, and obese humans appear to have reduced D2R-binding sites in their brain. Therefore, we hypothesized that spontaneous PRL release is enhanced in obese humans. To evaluate this hypothesis, we measured 24-h plasma PRL concentrations at 10-min intervals in 11 obese premenopausal women [body mass index (BMI), 33.3 +/- 0.7 kg/m(2)] and 10 lean premenopausal women of similar age (BMI, 21.2 +/- 0.6 kg/m(2)). Total body fat was determined using dual energy x-ray absorptiometry, and sc and visceral fat area was measured by magnetic resonance imaging in 10 obese subjects. PRL secretion rate was estimated by deconvolution analysis. All subjects were studied in the early follicular stage of their menstrual cycle. PRL secretion was significantly enhanced in obese women (total daily release, 137 +/- 8; lean controls, 92 +/- 8 microg/liter.24 h; P = 0.001) in proportion to their BMI (r(2) = 0.55; P < 0.001). Interestingly, PRL release was particularly associated with the size of the visceral fat mass (total PRL secretion vs. visceral fat area, r(2) = 0.64; P = 0.006). These data show that spontaneous PRL release is considerably enhanced in obese women in proportion to the size of their visceral fat mass. Because PRL is inhibited by D2R activation we speculate that elevated PRL secretion may be due to reduced D2R availability in the brain. PMID- 15356046 TI - Familial leucine-sensitive hypoglycemia of infancy due to a dominant mutation of the beta-cell sulfonylurea receptor. AB - Familial leucine-sensitive hypoglycemia of infancy was described in 1956 as a condition in which symptomatic hypoglycemia was provoked by protein meals or the amino acid, leucine. The purpose of this study was to determine the genetic basis for hypoglycemia in a family diagnosed with leucine-sensitive hypoglycemia in 1960. Recently diagnosed family members showed a dominantly transmitted pattern of diazoxide-responsive hyperinsulinism (HI). However, they did not fit the characteristics of HI caused by glutamate dehydrogenase gene mutations, previously felt to explain leucine-sensitive hypoglycemia. Islet function was examined using acute insulin response (AIR) tests to calcium, leucine, glucose, and tolbutamide as well as oral protein tolerance tests. Five of five affected family members showed an abnormal positive calcium AIR, and two of five showed a positive leucine AIR. Protein-induced hypoglycemia was demonstrated in five of six affected subjects. Mutation analysis of four known HI genes (sulfonylurea receptor 1, Kir6.2, glutamate dehydrogenase, and glucokinase) in family members identified an R1353H missense mutation in exon 33 of SUR1. (86)Rb(+) efflux and electrophysiological studies of R1353H SUR1 coexpressed with wild-type Kir6.2 in COSm6 cells demonstrated partially impaired ATP-dependent potassium channel function. Leucine-sensitive hypoglycemia in this family was found to result from a dominantly expressed SUR1 mutation. PMID- 15356047 TI - Postprandial ghrelin is elevated in black compared with white women. AB - Ghrelin, a gut-brain peptide that signals hunger, is normally suppressed after meals. Subnormal suppression of postprandial ghrelin, previously noted in obese, insulin-resistant individuals, may contribute to increased food intake. Given the ethnic disparities in obesity and obesity-related cardiovascular morbidity in the United States, the present study compared a single postprandial ghrelin measure in 43 women (22 white, 21 black). Each completed a rigorously controlled 4-d dietary intervention designed to maintain weight and constant daily sodium and potassium intake (220 mEq Na, 40 mEq K). Two hours after consuming a test meal of identical content, blood samples were drawn to assess postprandial ghrelin, leptin, and norepinephrine; resting cardiovascular function was measured; and a 24-h urinary cortisol sample was obtained. Independent of body mass index, postprandial ghrelin was significantly higher in black vs. white women, and higher ghrelin was associated with higher cortisol in blacks, who failed to show the expected inverse relation between ghrelin and central obesity seen in whites. Higher ghrelin was correlated with higher blood pressure but lower norepinephrine in obese women. These findings suggest subnormal postprandial ghrelin suppression (or faster ghrelin rebound) in black women, especially the obese, that might play a role in their increased prevalence of obesity and cardiovascular disorders. PMID- 15356048 TI - Increased day 15-17 serum pro-alphaC inhibin levels specific to successful pregnancy. AB - In early pregnancy, serum levels of the luteal-derived hormone pro-alphaC inhibin peak by the second week after conception. Whether this early rise is biologically important and a consistent feature of only successful pregnancy is unknown. We undertook a prospective cross-sectional study to determine whether serum pro alphaC inhibin levels at d 15-17 are predictive of a successful clinical in vitro fertilization pregnancy and compared levels between fresh embryo transfer (ET) and frozen-thawed ET. Median (95% confidence interval) pro-alphaC inhibin levels were 68 (57-76) pg/ml in 204 women who did not become clinically pregnant after ET, significantly lower than in either 90 women who became clinically pregnant after fresh ET and who had 3139 (1684-4220) pg/ml or in 39 women with a successful frozen ET who had 877 (678-1111) pg/ml. Pro-alphaC was highly sensitive and specific in predicting clinical pregnancy success but did not improve on the performance of human chorionic gonadotropin. Pro-alphaC inhibin levels were not correlated with progesterone or human chorionic gonadotropin. Levels were no higher in singleton compared with multiple pregnancies and did not increase across gestation, suggestive of a luteal source. The increase in circulating pro-alphaC inhibin in very early pregnancy is highly specific to clinical pregnancy, suggesting a possible biological role in early gestation. PMID- 15356049 TI - Is thyroid hormone suppression therapy prothrombotic? AB - The purpose of this study was to determine whether chronic thyroid hormone suppression therapy (THST) is prothrombotic. We obtained blood samples from 14 thyroid cancer patients while on THST and after they had become hypothyroid for radioiodine whole-body scanning and therapy. Prothrombin fragment 1 + 2, fibrinogen, factor VIII, antithrombin, tissue plasminogen activator antigen (tPA), plasminogen activator inhibitor 1 (PAI-1), PAI-1/tPA, and C-reactive protein were significantly (P < 0.05) higher in the hyper- than in the hypothyroid state, whereas protein C and plasmin-antiplasmin complexes were significantly lower during the hyperthyroid period. When the 10 female patients were hyperthyroid, their levels of prothrombin fragment 1 + 2, fibrinogen, protein S, antithrombin, tPA, PAI-1, and PAI-1/tPA were significantly higher (P 0.05). These results suggested that HLA-DR3 and CTLA-4 conferred a general increased risk for GD in both the sporadic and familial forms, and that the risk conferred by them was additive. However, HLA-DR3 and CTLA-4 did not have a stronger effect in the familial GD patients, suggesting that additional genes must contribute to the aggregation of GD within families. PMID- 15356064 TI - Evidence of early ovarian aging in fragile X premutation carriers. AB - Up to 28% of female fragile X premutation carriers develop premature ovarian failure. To test the hypothesis that fragile X premutation carriers with ovulatory menstrual cycles exhibit hormone changes characteristic of early ovarian aging, 11 regularly cycling fragile X premutation carriers, 24-41 yr old (34.5 +/- 5.7 yr, mean +/- sd), drew daily blood samples across one menstrual cycle. LH, FSH, estradiol, progesterone (P4), inhibin A, and inhibin B levels were compared with levels in 22 age-matched, regularly cycling women, 23-41 yr old (34.6 +/- 5.8 yr), at each cycle stage. Total cycle (26.1 +/- 1.0 vs. 28.2 +/ 0.4 d; P < 0.05) and follicular phase length (12.9 +/- 0.8 vs. 14.5 +/- 0.4 d; P < 0.05) were decreased in fragile X premutation carriers compared with age matched controls, whereas luteal phase length was similar (13.2 +/- 0.5 vs. 13.7 +/- 0.3 d; P = not significant). FSH was elevated across the follicular (21.9 +/- 3.5 vs. 11.2 +/- 0.5 IU/liter; P < 0.001) and luteal phases (14.6 +/- 3.9 vs. 7.9 +/- 0.5 IU/liter; P < 0.05) in fragile X premutation carriers compared with age matched controls. Inhibin B in the follicular phase (77 +/- 11 vs. 104 +/- 6 pg/ml; P < 0.05) and inhibin A (3.4 +/- 0.7 vs. 5.8 +/- 0.5 IU/ml; P < 0.01) and P4 [7.3 +/- 1.0 vs. 10.1 +/- 0.7 ng/ml (23.2 +/- 3.0 vs. 32.1 +/- 2.3 nmol/liter); P < 0.05] in the luteal phase were decreased in fragile X premutation carriers compared with age-matched controls, whereas there was no difference in estradiol or LH. In summary, despite regular ovulatory cycles, FSH was increased in fragile X premutation carriers compared with age-matched controls. The increased FSH was accompanied by decreased inhibin B in the follicular phase and inhibin A and P4 in the luteal phase. These hormonal changes suggest that fragile X premutation carriers exhibit early ovarian aging despite regular menstrual cycles. Early ovarian aging in fragile X premutation carriers likely results from decreased follicle number and function, as reflected by lower inhibin B, inhibin A, and P4 levels. PMID- 15356065 TI - AMP-activated protein kinase is not down-regulated in human skeletal muscle of obese females. AB - Obesity in humans is associated with lipid accumulation in skeletal muscle, insulin and leptin resistance, and type 2 diabetes. AMP-activated protein kinase (AMPK) is an important regulator of fatty acid (FA) metabolism in skeletal muscle. To address the hypothesis that lipid accumulation in skeletal muscle of obese subjects may be due to down-regulation of AMPK, we measured mRNA and protein levels of AMPK isoforms, AMPKalpha1 and -alpha2 activity, AMPK kinase activity, acetyl-coenzyme A carboxylase (ACCbeta) expression and phosphorylation, and FA metabolism in biopsies of rectus abdominus muscle from lean and obese women. We also examined the effect of 5-aminoimidazole-4-carboxamide riboside (AICAR) on AMPK activity and the effects of AICAR and leptin on FA metabolism. Skeletal muscle of obese subjects had increased total FA uptake and triglyceride esterification, and leptin failed to stimulate FA oxidation. However, AMPK mRNA and protein expression, AMPKalpha1 and -alpha2 activities, AMPK kinase activity, ACCbeta phosphorylation, and FA oxidation were similar in lean and obese subjects. Moreover, AICAR increased AMPKalpha2 activity, ACCbeta phosphorylation, and palmitate oxidation to a similar degree in muscle from lean and obese subjects. We conclude that the abnormal lipid metabolism and leptin resistance of skeletal muscle of obese subjects is not due to down-regulation of AMPK. In addition, the similar stimulation by AICAR of AMPK in skeletal muscle of lean and obese subjects suggests that direct pharmacological activation of AMPK may be a therapeutic approach for stimulating FA oxidation in the treatment of human obesity. PMID- 15356066 TI - Activation of somatostatin-receptor subtype-2/-5 suppresses the mass, frequency, and irregularity of growth hormone (GH)-releasing peptide-2-stimulated GH secretion in men. AB - Somatostatin antagonizes the stimulatory actions of GHRH and GH-releasing peptides (GHRPs). However, precisely how the inhibitory susceptibilities of the two secretagogues differ is not clear. One interpretative difficulty is that native somatostatin activates six different receptor subtypes. The present study adopts the complementary strategy of enforcing feedback inhibition via the preferential somatostatin receptor subtype 2 and 5 (SSTR-2/-5) agonist, octreotide. We postulated that putative SSTR-2/-5 agonism would unmask secretagogue-selective interactions in the control of GH secretory burst mass, frequency, and/or regularity. To this end, 10 healthy men each underwent eight randomly ordered, separate-day, fasting morning infusion sessions. Interventions comprised sc administration of octreotide (1 microg/kg), followed by bolus iv injection of saline, GHRH (1 microg/kg), GHRP-2 (1 microg/kg), or both peptides. Compared with placebo, the SSTR-2/-5 agonist reduced fasting GH concentrations from 0.27 +/- 0.07 to 0.12 +/- 0.02 microg/liter (P = 0.020), GH secretory burst mass from 2.7 +/- 0.65 to 0.55 +/- 0.11 microg/liter (P = 0.013), and basal GH secretion from 0.24 +/- 0.043 to 0.11 +/- 0.015 microg/liter.100 min (P = 0.0063). The foregoing outcomes were selective, because octreotide did not alter GH secretory burst frequency (3.1 +/- 0.5 vs. 3.3 +/- 0.21 events/3 h) or the regularity of the GH release process (approximate entropy, 0.58 +/- 0.048 vs. 0.68 +/- 0.064). In the GHRP-2-stimulated setting, presumptive SSTR-2/-5 agonism suppressed all three GH secretory burst masses, from 28 +/- 3.2 to 18 +/- 2.0 (P = 0.045); GH pulse frequency, from 3.3 +/- 0.30 to 2.0 +/- 0.18 (P = 0.0025); and the irregularity (approximate entropy) of GH release, from 0.648 +/- 0.049 to 0.433 +/- 0.047 (P < 0.01). In contrast, in the GHRH and combined GHRH/GHRP-2 stimulated contexts, octreotide decreased only GH secretory burst mass (P = 0.047). In summary, the present data indicate that GH secretory burst mass, frequency, and orderliness are subject to interactive control by at least SSTR-2/ 5-dependent feedback and GHRP-dependent feedforward signals. PMID- 15356067 TI - Early impairment of endothelial structure and function in young normal-weight women with polycystic ovary syndrome. AB - The aim of this study was to evaluate the presence of early vascular damage in young normal-weight women with polycystic ovary syndrome (PCOS). Thirty young normal-weight women with PCOS, who had no additional metabolic or cardiovascular diseases, and 30 healthy women (controls) matched for age and body mass index were studied. A complete hormonal assay was performed in each subject. Serum insulin and glucose levels were measured at baseline and after the oral glucose tolerance test. Plasma endothelin-1 levels and serum lipid profile were also assessed. The endothelial function was studied by flow-mediated dilation on the brachial artery, and arterial structure was evaluated by intima-media thickness measurement using Doppler ultrasound of both common carotid arteries.A significant (P < 0.05) difference in flow-mediated dilation (14.3 +/- 1.9% vs. 18.1 +/- 2.0% for PCOS patients and controls, respectively) and in intima-media thickness (0.53 +/- 0.09 mm vs. 0.39 +/- 0.08 mm for PCOS patients and controls, respectively) was found between PCOS and control subjects. Serum endothelin-1 levels were also significantly (P < 0.05) higher in PCOS patients compared with controls (1.1 +/- 0.4 pmol/liter vs. 0.5 +/- 0.2 pmol/liter for PCOS patients and controls, respectively).In conclusion, our data show that young, normal-weight, nondyslipidemic, nonhypertensive women with PCOS have an early impairment of endothelial structure and function. PMID- 15356068 TI - Tibolone activates nitric oxide synthesis in human endothelial cells. AB - After the unexpected findings of the Women's Health Initiative trial, indicating that traditional cardiovascular risk markers fail to predict the effects of hormone replacement therapy, it is of interest to characterize how steroids act on vascular cells. This is particularly important for tissue-specific drugs such as tibolone, whose actions may differ from other preparations. Because nitric oxide (NO) is a key regulator of vascular tone and atherogenesis, we studied its regulation by tibolone and its metabolites on human endothelial cells. Tibolone and its estrogenic metabolites (3alpha- and 3beta-OH tibolone) activate NO synthesis by recruiting functional estrogen receptors, whereas the progestogenic/androgenic metabolite (Delta(4) isomer) has no effect. During prolonged exposures, tibolone and the estrogenic compounds enhance the expression of endothelial NO synthase (eNOS). In addition, tibolone is able to induce rapid activation of eNOS, leading to rapid increases in the release of NO. Relevant for its clinical effects, the sulfated metabolites of tibolone are also effective in activating eNOS. Different from estrogen, rapid activation of eNOS does not rely on recruitment of phosphatidylinositol-3 kinase but rather on MAPK-dependent cascades. These results help to understand the mechanisms of action of tibolone on the cardiovascular system and have relevant clinical implications. PMID- 15356069 TI - Age-dependent regulation of lipogenesis in human and rat adipocytes. AB - The regulation of adipocyte metabolism is of importance for adipose tissue growth and therefore also for the development of obesity. This study was designed to investigate the regulation of basal and insulin-induced lipogenesis, glucose transport, and glucose transporter protein expression in human and rat adipocytes from different age groups. The study included 21 infants, 21 children, nine adults, and 80 male weaned and 20 male adult Fischer rats. The lipogenesis experiments were performed under conditions at which glucose transport is rate limiting. Basal lipogenesis was approximately three times higher in infants and children than in adults, whereas insulin-induced lipogenesis was two times higher in infants than in children and adults. In rats, basal lipogenesis, insulin induced lipogenesis, and insulin sensitivity were two times higher in weaned than in adult animals. Moreover, basal and insulin-induced glucose transport were two times higher in weaned than in adult rats. No differences were detected in GLUT1 or GLUT4 content between any of the age groups in human or in rat adipocytes. In conclusion, basal and insulin-stimulated lipogenesis are increased in adipocytes early in life. This may promote adipose tissue growth in early age. The data indicate that age-dependent variation in basal and insulin-stimulated lipogenesis is differently regulated. PMID- 15356070 TI - Triglyceride hydrolase activities and expression of fatty acid binding proteins in the human placenta in pregnancies complicated by intrauterine growth restriction and diabetes. AB - Triglyceride (TG) hydrolases in the placental microvillous plasma membrane (MVM) release fatty acids from circulating lipoproteins and represent the critical initial step in transplacental fatty acid transfer. We investigated the activity of two TG hydrolases in MVM isolated from placentas of appropriately grown for gestational age pregnancies and pregnancies complicated by intrauterine growth restriction (IUGR), insulin-dependent diabetes mellitus (IDDM) or gestational diabetes mellitus (GDM). In addition, we measured protein expression of lipoprotein lipase (LPL) in MVM and two fatty acid binding proteins (L- and C FABP) in placental homogenates. The TG hydrolase activities were assessed by measuring hydrolysis of (3)H-trioleic acid incorporated into intralipid micelles after incubation with MVM. The placenta-specific TG hydrolase activity (optimum at pH 6) did not differ in the patient groups studied. MVM LPL activity (optimum at pH 8) was reduced by 47% in preterm IUGR (n = 8, P < 0.05), compared with gestational age-matched controls. The LPL activity in placentas of IDDM pregnancies was increased by 39% (n = 8, P < 0.05), compared with controls. No significant differences were observed in cases of GDM. We found no alteration in protein expression of LPL or C-FABP. The expression of L-FABP was increased by 112% (n = 8, P < 0.05) in IDDM and 64% (n = 8, P < 0.05) in GDM. These results indicate that alterations in MVM LPL activity and expression of L-FABP may contribute to the altered lipid deposition and metabolism in IUGR and diabetic pregnancies. PMID- 15356071 TI - Hormone therapy impairs endothelial function in postmenopausal women with type 2 diabetes mellitus treated with rosiglitazone. AB - Diabetes and ovarian senescence are associated with impaired endothelial function and altered arterial mechanical properties. Alterations in normal vascular structure and functioning are the primary cause of mortality and morbidity with type 2 diabetes. Similarly, after menopause, women experience an increase in the rate of cardiovascular disease. Thiazolidinediones have exhibited a number of antiatherogenic actions in populations with type 2 diabetes. The effect of thiazolidinediones in combination with hormone therapy (HT) in postmenopausal women is, however, unknown. To assess whether HT (transdermal estradiol 50 microg and micronized progesterone (100 mg/d) affects vascular function, 21 women receiving rosiglitazone were randomly assigned to receive HT or placebo for 12 wk in a double-blind crossover design. Measures of glycemic control, lipids, blood pressure, flow-mediated dilation, and distensibility index were undertaken at baseline and after each treatment. As a result, flow-mediated dilation was significantly reduced (15.3 +/- 3.8 to 6.6 +/- 1.6%, P = 0.02) with HT, whereas lipids, blood pressure, and distensibility index were unchanged. Placebo had no significant affect on any variables. Thus, the addition of HT to rosiglitazone treatment attenuates endothelial function without altering other cardiovascular risk factors. Caution should, therefore, be exercised when considering combined treatment with thiazolidinedione and HT. PMID- 15356072 TI - Adiponectin is related to CD146, a novel marker of endothelial cell activation/injury in chronic renal failure and peritoneally dialyzed patients. AB - Adiponectin has antiatherogenic properties and attenuates endothelial inflammatory responses. CD146 is a novel cell adhesion molecule localized at the endothelial junction. In renal failure, endothelial dysfunction and atherosclerosis are almost universal. We studied possible correlations between adiponectin, CD146, and other markers of endothelial cell injury in patients with chronic renal failure (CRF) on conservative treatment and patients with and without diabetic nephropathy maintained on chronic ambulatory peritoneal dialysis (CAPD). We assessed adiponectin, tissue factor pathway inhibitor (TFPI), plasminogen activator inhibitor (PAI-1), thrombin-activatable fibrinolysis inhibitor, and endothelial function/injury markers: von Willebrand factor, thrombomodulin, vascular cell adhesion molecule (VCAM), intercellular adhesion molecule, and CD146. Adiponectin was elevated in patients with CRF and on CAPD. It correlated significantly, with PAI-1, thrombin-activatable fibrinolysis inhibitor, intercellular adhesion molecule, VCAM, and CD146 in nondiabetics on CAPD. In diabetics, CAPD adiponectin correlated positively with C146 and VCAM and negatively with PAI and TFPI. In multivariate regression analysis, only CD146 remained a positive predictor of adiponectin in all CAPD patients. In CRF, adiponectin correlated with CD146. In healthy volunteers, adiponectin correlated with TFPI and CD146. Elevated adiponectin related to CD146 may be the expression of a counterregulatory response aimed at mitigating the consequences in endothelial damage and increased cardiovascular risk in renal failure. PMID- 15356073 TI - Studies on the origin of circulating 18-hydroxycortisol and 18-oxocortisol in normal human subjects. AB - 18-Hydroxycortisol (18-OHF) and 18-oxocortisol (18-oxoF) are derivatives of cortisol found in primary aldosteronism but whose origin and regulation in normal subjects are uncertain. 18-OHF can be synthesized by zona fasciculata 11-beta hydroxylase; 18-oxoF can only be produced by zona glomerulosa aldosterone synthase (AS). Stably transfected cell lines expressing either CYP11B1 (11beta hydroxylase) or CYP11B2 (AS) were incubated with cortisol and other substrates over a range of concentrations. Both enzymes could synthesize 18-OHF from cortisol, but only AS could synthesize 18-oxoF. AS was more efficient than 11beta hydroxylase at 18-hydroxylation. The apparent Michaelis-Menten constant (K(m)) of AS for cortisol was estimated to be 2.6 microm. In five patients with adrenal insufficiency maintained on hydrocortisone, urinary free cortisol and cortisone levels were high; 18-oxoF was detectable in all patients and 18-OHF in three. It is likely that the 18-oxygenated steroids were synthesized from circulating cortisol, either in the zona glomerulosa or at extraadrenal sites. In eight male volunteers, dexamethasone treatment decreased urinary excretion rates of free cortisol, cortisone, 18-OHF, and 18-oxoF, confirming dependence of 18-oxygenated steroid levels on cortisol availability. In both groups, hydrocortisone administration resulted in detectable levels of 18-OHF and raised levels of 18 oxoF. There was close correlation between 18-oxoF and cortisol excretion during hydrocortisone administration in normal subjects (r = 0.86; P < 0.001). These data show, for the first time, that 18-OHF and 18-oxoF can be synthesized from circulating cortisol. The close correlation between 18-oxoF and cortisol suggests that 18-oxoF is normally produced by the action of AS using circulating cortisol as a substrate. Although 18OHF can be synthesized using circulating cortisol as substrate, our data suggest this is normally produced in the zona fasciculata by 11beta-hydroxylase from locally available cortisol. PMID- 15356074 TI - Dose-dependent effects of recombinant human insulin-like growth factor (IGF) I/IGF binding protein-3 complex on overnight growth hormone secretion and insulin sensitivity in type 1 diabetes. AB - GH hypersecretion in type 1 diabetes has been implicated in the pathogenesis of insulin resistance, and microangiopathic complications, and may result from reduced circulating IGF levels. We examined the effects of recombinant human (rh)IGF-I [complexed in equimolar ratio with rhIGF binding protein (BP)-3 (rhIGF I/IGFBP-3)] replacement on overnight GH levels and insulin sensitivity in type 1 diabetes. Fifteen subjects, 13-24 yr old (10 male), were given rhIGF-I/IGFBP-3 or placebo as a daily sc injection for 2 d. After the second injection overnight, insulin requirements for euglycemia were determined (0400-0800 h), followed by a 4-h, two-step (insulin, 0.6 and 1.5 mU/kg.min) hyperinsulinemic euglycemic [90 mg/dl (5 mmol/liter)] clamp. In each subject, the protocol was repeated on three occasions in random order. Seven subjects received placebo and rhIGF-I/IGFBP-3 (0.1 mg/kg.d and 0.4 mg/kg.d), and eight subjects received placebo and rhIGF I/IGFBP-3 (0.2 mg/kg.d and 0.8 mg/kg.d). We found dose-dependent increases in circulating IGF-I and IGFBP-3 concentrations after rhIGF-I/IGFBP-3. These were paralleled by significant reductions in mean overnight GH levels and GH pulse amplitude. We also observed dose-dependent effects of rhIGF-I/IGFBP-3 on overnight insulin requirements for euglycemia, with reductions of up to 41%. Insulin sensitivity, defined by M-values, was improved with rhIGF-I/IGFBP-3 (0.4 and 0.8 mg/kg.d). Thus, restoration of circulating IGF-I and IGFBP-3 levels with rhIGF-I/IGFBP-3 suppresses GH secretion in adolescents with type 1 diabetes, leading to reduced insulin requirements and improvements in insulin sensitivity. PMID- 15356075 TI - Intranasal atrial natriuretic peptide acts as central nervous inhibitor of the hypothalamo-pituitary-adrenal stress system in humans. AB - Increased hypothalamo-pituitary-adrenal activity contributes to morbidity in widespread metabolic and psychiatric diseases. Inhibition of hypercortisolism represents a promising therapeutic strategy in these conditions, which currently cannot be used. Here, we tested the hypothesis that atrial natriuretic peptide (ANP) administered intranasally is a safe and feasible inhibitor of pituitary adrenal activity at the central nervous level. Thirty minutes after intranasal administration of ANP (1 mg) and placebo, pituitary-adrenal activity was stimulated in 18 healthy men by two tests: 1) a standard insulin-hypoglycemia test and 2) CRH combined with vasopressin (VP), respectively. ACTH, cortisol, VP, blood pressure, heart rate, and measures of fluid balance were also recorded. Pretreatment with ANP suppressed cortisol (P < 0.01) and ACTH (P < 0.05) secretory responses to insulin-induced hypoglycemia to about half of that seen after placebo, but pituitary-adrenal activity was not suppressed by CRH/VP injection (P > 0.7). Indicators of fluid balance, cardiovascular parameters, and self-report measures were not influenced by the treatment. Results indicate a strong inhibition of stimulated pituitary-adrenal activity after intranasal administration of ANP. The absence of an effect on CRH/VP-induced pituitary adrenal responses suggests a direct action of the peptide on the central nervous system inhibiting stimulated hypothalamo-pituitary-adrenal activity at the hypothalamic level. PMID- 15356076 TI - Sustained reduction in plasma free fatty acid concentration improves insulin action without altering plasma adipocytokine levels in subjects with strong family history of type 2 diabetes. AB - To investigate the effect of a sustained (7-d) decrease in plasma free fatty acid (FFA) concentration in individuals genetically predisposed to develop type 2 diabetes mellitus (T2DM), we studied the effect of acipimox, a potent inhibitor of lipolysis, on insulin action and adipocytokine concentrations in eight normal glucose-tolerant subjects (aged 40 +/- 4 yr, body mass index 26.5 +/- 0.8 kg/m(2)) with at least two first-degree relatives with T2DM. Subjects received an oral glucose tolerance test (OGTT) and 120 min euglycemic insulin clamp (80 mU/m(2).min) with 3-[(3)H] glucose to quantitate rates of insulin-mediated whole body glucose disposal (Rd) and endogenous (primarily hepatic) glucose production (EGP) before and after acipimox, 250 mg every 6 h for 7 d. Acipimox significantly reduced fasting plasma FFA (515 +/- 64 to 285 +/- 58 microm, P < 0.05) and mean plasma FFA during the OGTT (263 +/- 32 to 151 +/- 25 microm, P < 0.05); insulin mediated suppression of plasma FFA concentration during the insulin clamp also was enhanced (162 +/- 18 to 120 +/- 15 microm, P < 0.10). Following acipimox, fasting plasma glucose (5.1 +/- 0.1 vs. 5.2 +/- 0.1 mm) did not change, whereas mean plasma glucose during the OGTT decreased (7.6 +/- 0.5 to 6.9 +/- 0.5 mm, P < 0.01) without change in mean plasma insulin concentration (402 +/- 90 to 444 +/- 102 pmol/liter). After acipimox Rd increased from 5.6 +/- 0.5 to 6.8 +/- 0.5 mg/kg.min (P < 0.01) due to an increase in insulin-stimulated nonoxidative glucose disposal (2.5 +/- 0.4 to 3.5 +/- 0.4 mg/kg.min, P < 0.05). The increment in Rd correlated closely with the decrement in fasting plasma FFA concentration (r = -0.80, P < 0.02). Basal EGP did not change after acipimox (1.9 +/- 0.1 vs. 2.0 +/- 0.1 mg/kg.min), but insulin-mediated suppression of EGP improved (0.22 +/ 0.09 to 0.01 +/- 0.01 mg/kg.min, P < 0.05). EGP during the insulin clamp correlated positively with the fasting plasma FFA concentration (r = 0.49, P = 0.06) and the mean plasma FFA concentration during the insulin clamp (r = 0.52, P < 0.05). Plasma adiponectin (7.1 +/- 1.0 to 7.2 +/- 1.1 microg/ml), resistin (4.0 +/- 0.3 to 3.8 +/- 0.3 ng/ml), IL-6 (1.4 +/- 0.3 to 1.6 +/- 0.4 pg/ml), and TNFalpha (2.3 +/- 0.3 to 2.4 +/- 0.3 pg/ml) did not change after acipimox treatment. We concluded that sustained reduction in plasma FFA concentration in subjects with a strong family history of T2DM increases peripheral (muscle) and hepatic insulin sensitivity without increasing adiponectin levels or altering the secretion of other adipocytokines by the adipocyte. These results suggest that lipotoxicity already is well established in individuals who are genetically predisposed to develop T2DM and that drugs that cause a sustained reduction in the elevated plasma FFA concentration may represent an effective modality for the prevention of T2DM in high-risk, genetically predisposed, normal glucose-tolerant individuals despite the lack of an effect on adipocytokine concentrations. PMID- 15356077 TI - Regulation of net hepatic glycogenolysis and gluconeogenesis during exercise: impact of type 1 diabetes. AB - The effects of type 1 diabetes on the contributions of net hepatic glycogenolysis and gluconeogenesis to glucose production (GP) at rest and during moderate (MOD) and high (HI) intensity running were examined in healthy control (n = 6) and type 1 diabetic (n = 5) subjects matched for age, weight, and maximum aerobic capacity by combined noninvasive measurements of hepatic glycogen content using (13)C nuclear magnetic resonance spectroscopy and determination of GP using [6,6 (2)H(2)]glucose. In the control subjects, GP increased in proportion to the intensity of the exercise [at rest (REST), 14.3 +/- 0.5; MOD, 18.1 +/- 0.9; HI, 28.8 +/- 1.3 micromol/(kg-min); P = 0.001, three-way comparison], and this was accounted for by an increase in the percent contribution of net hepatic glycogenolysis to GP (REST, 32 +/- 1%; MOD, 49 +/- 5%; HI, 57 +/- 5%; P = 0.006). In the diabetic subjects, resting rates of GP were 60% higher than those in the control subjects (P < 0.0001) and increased in proportion to the workload. In contrast, the contributions of net hepatic glycogenolysis to GP were consistently lower than those in the control subjects (REST, 20 +/- 6%; MOD, 32 +/- 13%; HI, 32 +/- 3%; P = 0.006 vs. control), and the exaggerated rates of GP could be entirely accounted for by increased rates of gluconeogenesis. In conclusion, 1) increases in GP in healthy control subjects with exercise intensity can be entirely attributed to increases in net hepatic glycogenolysis. 2) In contrast, moderately controlled type 1 diabetic subjects exhibit increased rates of GP both at rest and during exercise, which can be entirely accounted for by increased gluconeogenesis. PMID- 15356078 TI - Insulin sensitivity following agent orange exposure in Vietnam veterans with high blood levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - Our objective was to determine whether insulin sensitivity was related to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) in Vietnam veterans exposed to Agent Orange. Air Force veterans of Operation Ranch Hand, the unit responsible for spraying Agent Orange and other herbicides in Vietnam from 1962 to 1971, and comparison veterans who did not spray herbicides were included. We measured insulin sensitivity (S(I)) using a frequently sampled iv glucose tolerance test in a matched study of 29 matched pairs of veterans and a quantitative insulin sensitivity check index (QUICKI) based on fasting glucose and insulin in 71 matched pairs. No group differences were found with regard to the mean values of S(I), QUICKI, TNFalpha, adiponectin, and two measures of insulin secretion. However, S(I) and QUICKI decreased significantly with regard to TCDD (P = 0.01 and 0.02). A corresponding pattern (although not significant) was found for blood levels of TNFalpha and adiponectin. These data suggest that high blood TCDD levels may promote an insulin-resistant state, but the magnitude of this effect appeared to be small, such that an 18-fold increase in blood TCDD due to increased exposure resulted in only a 10% change in S(I) in the 29 matched pairs. PMID- 15356079 TI - The measurement of maternal plasma corticotropin-releasing factor (CRF) and CRF binding protein improves the early prediction of preeclampsia. AB - In the present study we measured maternal plasma concentrations of two placental neurohormones, corticotropin-releasing factor (CRF) and CRF-binding protein (CRF BP), in 58 at-risk pregnant women consecutively enrolled between 28 and 29 wk of pregnancy to evaluate whether their evaluation may predict third trimester-onset preeclampsia (PE). The statistical significance was assessed by t test. The cut off points for defining altered CRF and CRF-BP levels for prediction of PE were chosen by receiving operator characteristics curve analysis, and the probability of developing PE was calculated for several combinations of hormone testing results. CRF and CRF-BP levels were significantly (both P < 0.0001) higher and lower, respectively, in the patients (n = 20) who later developed PE than in those who did not present PE at follow-up. CRF at the cut-off 425.95 pmol/liter achieved a sensitivity of 94.8% and a specificity of 96.9%, whereas CRF-BP at the cut-off 125.8 nmol/liter combined a sensitivity of 92.5% and a specificity of 82.5% as single markers for prediction of PE. The probability of PE was 34.5% in the whole study population, 93.75% when both CRF and CRF-BP levels were changed, and 0% if both hormone markers were unaltered. The measurement of CRF and CRF-BP levels may add significant prognostic information for predicting PE in at-risk pregnant women. PMID- 15356080 TI - Reversal of insulin resistance postpartum is linked to enhanced skeletal muscle insulin signaling. AB - The restoration of maternal insulin sensitivity postpartum represents an important physiological and metabolic adaptation in a woman's reproductive lifespan. The present study was conducted to examine the potential cellular mechanisms underlying the changes in insulin sensitivity from late pregnancy to postpartum in human skeletal muscle. Nine nonobese women (age, 32 +/- 2 yr; body mass index, 21.2 +/- 0.8 kg/m(2)) with normal glucose tolerance were studied during late pregnancy (30-36 wk) and again approximately 1 yr postpartum using a euglycemic-hyperinsulinemic clamp (5 mm glucose, 40 mU/m(2).min insulin) to determine insulin sensitivity. Biopsies of the vastus lateralis muscle were obtained in the basal state before each clamp. Insulin sensitivity improved by 74% from late pregnancy to 1 yr postpartum (5.5 +/- 0.6 vs. 9.6 +/- 0.9 mg/kg fat free mass.min; P < 0.005). Skeletal muscle insulin receptor (IR) protein expression increased by 42% postpartum, as measured by ELISA (4.0 +/- 0.6 vs. 5.7 +/- 0.6 ng/g protein; P < 0.05) and by Western blotting of the IR beta-subunit (28.7 +/- 4.7 vs. 42.0 +/- 4.8 arbitrary units; P < 0.003). However, in vitro studies showed that when adjusted for IR concentration, maximal insulin stimulated (100 nm) IR tyrosine phosphorylation (0.75 +/- 0.06 vs. 0.92 +/- 0.08 U) and IR tyrosine kinase activity (183.8 +/- 27.0 vs. 204.3 +/- 23.7 fmol ATP/ng IR) were unchanged. There was a 69% increase in IR substrate-1 (IRS-1) protein expression (P = 0.05) in muscle postpartum. In addition, the p85alpha regulatory subunit of phosphatidylinositol 3-kinase was markedly reduced by 55% (P < 0.02) postpartum. The change in insulin sensitivity from late pregnancy to postpartum correlated highly with the corresponding change in IRS-1 protein (r = 0.84; P < 0.007). Downstream signaling proteins, including total Akt and p70s6 kinase, and the glucose transporter protein GLUT-4, were similar at both time points. These data suggest that reduced IR tyrosine kinase activity is not a major factor in the IR of pregnancy in lean women with normal glucose tolerance. Rather, the reversal of insulin resistance 1 yr postpartum is accompanied by increased skeletal muscle IRS-1 along with a down-regulation of the p85alpha subunit of phosphatidylinositol 3-kinase. These changes may allow for greater p85/p110 binding to IRS-1 and play a significant physiological role in the underlying metabolic adaptation to normal human pregnancy and restoration of insulin sensitivity postpartum. PMID- 15356081 TI - Estrone sulfate is a major source of local estrogen formation in human bone. AB - Estrone sulfate (E1S) is the most abundant estrogen in the circulation of adults. The present study was undertaken to assess estrone (E1) and estradiol formation from E1S in freshly resected bone [bone fragments (BFs)] and osteoblast-like cells (hOB) cultured from BFs. Furthermore, we compared estrogen formation from E1S in rat and human osteosarcoma (OS) cell lines and that of estrogen formation from E1S with that of aromatization of androstenedione and testosterone in BFs and those from E1S and androstenedione in hOB cells. The bone used was from the head of the femur from a total of 15 women and 12 men. Steroid sulfatase activity (STA) was found, and the formation of estrone and estradiol from E1S was demonstrated. STA was similar in cells derived from BFs of men and women. STA was significantly lower in OS cell lines, compared with hOB cells. Estrogen formation from E1S in BFs was at least 20 times higher than that from androstenedione and about 50 times higher than that from testosterone. Similarly, estrogen formation from E1S in hOB cells exceeded the values derived from aromatization of androstenedione by two orders of magnitude. Based on these results, we conclude that hOB cells express the same pattern of E1S metabolism as resected bone and thus may accurately mirror the in vivo situation in man. In comparison with hOB cells, STA is fundamentally lower in widely used OS cell lines that express an osteoblastic phenotype. This shortcoming precludes their use as model cell lines to unravel STA metabolic pathways and its regulation in nontumorous bone. E1S is a major source of local bioactive estrogen formation in human bone. Because bone is highly susceptible to estrogen action, local estrogen formation from E1S may play an important role in bone maturation and homeostasis, particularly in elderly adults. PMID- 15356082 TI - Combined hyperinsulinemia and hyperglycemia, but not hyperinsulinemia alone, suppress human skeletal muscle lipolytic activity in vivo. AB - Effects of circulating insulin and glucose concentrations on skeletal muscle and adipose tissue lipolytic activity were investigated in 10 type 1 diabetes patients with no endogenous insulin secretion. Microdialysis measurements of interstitial glycerol and determination of fractional glycerol release were carried out during standardized combinations of relative hypoinsulinemia/moderate hyperglycemia (11 mmol/liter), hyperinsulinemia/ normoglycemia (5 mmol/liter), and hyperinsulinemia/moderate hyperglycemia, respectively. Local tissue blood flow rates were measured with the (133)Xe clearance technique. In response to the change from hypo- to hyperinsulinemia, the fractional release of glycerol decreased from 159.6 +/- 17.8 to 85.1 +/- 13.7 micromol/liter (P < 0.0001) in adipose tissue, whereas it remained unchanged in skeletal muscle (44.6 +/- 6.4 vs. 36.0 +/- 7.4 micromol/liter; not significant). When hyperinsulinemia was combined with hyperglycemia, fractional glycerol release was further reduced in adipose tissue (64.5 +/- 12.2 micromol/liter; P < 0.05), and in this situation it was also markedly decreased in skeletal muscle (18.1 +/- 4.8 micromol/liter; P < 0.0001). Skeletal muscle blood flow was unaltered over the respective study periods. Adipose tissue blood flow decreased by 50% in response to hyperinsulinemia (P < 0.0005), but no further change was seen when hyperinsulinemia was combined with hyperglycemia. It is concluded that in patients with type 1 diabetes, insulin does not exert an antilipolytic effect in skeletal muscle during normoglycemia. However, in response to combined hyperinsulinemia and hyperglycemia, the lipolytic activity in skeletal muscle is restrained in a similar way as in adipose tissue. This may be explained by a glucose-mediated potentiation of the antilipolytic effectiveness of insulin. PMID- 15356083 TI - Responses of lipolysis and salivary cortisol to food intake and physical activity in lean and obese children. AB - This investigation was conducted to determine whether there were differences in lipolytic responses to feeding and physical activity between lean (LN) and obese (OB) children, and if these responses were related to cortisol. Fourteen LN and 11 OB children participated in this study of abdominal lipolysis and salivary cortisol response to breakfast and lunch with an intervening exercise session. Calculated fasting glycerol release was lower in OB than LN (0.645 +/- 0.06 vs. 0.942 +/- 0.11 micromol/ml; P < 0.05). Fasting adipose tissue nutritive flow was lower in OB than in LN subjects, but responses to feeding and exercise were not different. Breakfast elicited a decrease in interstitial glycerol concentration in LN (-33%; P < 0.05), but not in OB (-5%), children, although decreases in glycerol concentration in response to lunch were similar (LN, -41%; OB, -36%). An interaction was evident in the salivary cortisol response to breakfast (LN, no change; OB, increase) and exercise (LN, no change; OB, decrease), but there were no group differences in response to lunch. Alterations in salivary cortisol and lipolysis were not related. These data suggest that salivary cortisol and lipolytic responses are not necessarily linked, but are altered in obesity. Furthermore, prior exercise may improve the antilipolytic response to a meal in OB children. PMID- 15356084 TI - Dehydroepiandrosterone increases endothelial cell proliferation in vitro and improves endothelial function in vivo by mechanisms independent of androgen and estrogen receptors. AB - Dehydroepiandrosterone (DHEA) may be beneficial in cardiovascular health, but mechanisms of DHEA action in the cardiovascular system are unclear. We have therefore 1) determined DHEA effects on the proliferation of cultured endothelial cells (EC), 2) compared effects of DHEA with estradiol (E) and testosterone (T), and 3) examined DHEA effects on subcellular messengers. We have in addition examined effects of DHEA (100 mg/d, 3 months) in 36 healthy postmenopausal women on blood pressure, lipids, and endothelial function, assessed noninvasively in large vessels by flow-mediated dilation of the brachial artery during reactive hyperemia, and in small vessels by laser Doppler velocimetry with iontophoresis of acetylcholine. DHEA, E, and T all increased EC proliferation; the effect of E was abolished by the estrogen receptor antagonist ICI 182,780, and that of T was abolished by the androgen receptor antagonist flutamide; neither blocked the effect of DHEA. In vitro, DHEA increased EC expression of endothelial nitric oxide synthase and activity of extracellular signal-regulated kinase 1/2. In vivo, DHEA increased flow-mediated dilation and laser Doppler velocimetry and reduced total plasma cholesterol. Thus, DHEA increases EC proliferation in vitro by mechanism(s) independently of either androgen receptor or estrogen receptor and in vivo enhances large and small vessel EC function in postmenopausal women. PMID- 15356085 TI - Flutamide-metformin plus ethinylestradiol-drospirenone for lipolysis and antiatherogenesis in young women with ovarian hyperandrogenism: the key role of early, low-dose flutamide. AB - A low-dose combination of flutamide-metformin and ethinylestradiol-drospirenone was recently found to reduce the excess of total and abdominal fat, to diminish the deficit in lean mass, and to attenuate the dysadipocytokinemia of young women with ovarian hyperandrogenism, a variant of polycystic ovary syndrome. We questioned the need to give flutamide, an androgen receptor blocker, together with an oral contraceptive that contains drospirenone, a progestin claimed to have antiandrogen properties. The additive effects of low-dose flutamide (62.5 mg/d) were assessed over 3 months in young patients with hyperinsulinemic ovarian hyperandrogenism (n = 40; age, approximately 17 yr; body mass index, approximately 22 kg/m(2)); all participants started on metformin (850 mg/d) and a fourth-generation contraceptive (ethinylestradiol 30 microg plus drospirenone 3 mg, 21 d/month), and they were randomized to receive flutamide in addition (n = 20) or not (n = 20). Fasting blood glucose, serum insulin, lipid profile, testosterone, adiponectin, and IL-6 were determined at baseline and after 3 months, together with body composition (by dual x-ray absorptiometry) and with Doppler assessment of ovarian arterial resistance. At start, the pulsatility and resistance indices of ovarian arteries were elevated. By comparison of 3-month changes between randomized subgroups, the addition of low-dose flutamide was found to have consistently (more) normalizing effects on low-density lipoprotein cholesterol, IL-6, and adiponectin, lean body mass, total and abdominal fat mass, and arterial flow in the ovaries. In conclusion, low-dose flutamide is herewith identified as a pivotal component within a first contraceptive combination therapy that has been shown to attenuate the hypoadiponectinemia, ovarian vascular hyperresistance, lean mass deficit, and central adiposity of young women with polycystic ovary syndrome. Finally, these data challenge any claim that drospirenone, as currently used in a contraceptive, is a clinically significant antiandrogen. PMID- 15356086 TI - The influence of lipoprotein lipase gene variation on postprandial lipoprotein metabolism. AB - Lipoprotein lipase (LPL) is one of the key enzymes in the metabolism of triacylglycerol-rich lipoproteins (TRL). We evaluated whether the association of LPL HindIII (H1/H2) and Serine447-Stop (S447X) polymorphisms may explain the interindividual variability observed during postprandial lipemia. Fifty-one healthy male volunteers (26 with the H2S447 genotype, 15 with the H1X447 genotype, and 10 with the H1S447 genotype) were subjected to a vitamin A-fat load test consisting of 1 g fat/kg body weight and 60,000 IU vitamin A. Blood was drawn every hour until the 6th hour and every 2 h and 30 min until the 11th hour. Data revealed that subjects that are homozygous for the H2 allele (H2H2) showed a higher postprandial response for small TRL, retinyl palmitate (RP), large TRL-RP, large TRL-B48, and small TRL-B48 levels. Furthermore, in the case of the S447X polymorphism, 447Ter carriers had a lower postprandial response for small TRL-RP, large TRL-B48, and small TRL-RP. Subjects with the LPL H2S447 genotype had higher plasma triacylglycerol, large TRL-triacylglycerol, large TRL-RP, small TRL-RP, and large TRL-B48 (P < 0.037) than H1X447 subjects. The modifications observed in postprandial lipoprotein metabolism in young normolipemic males with LPL polymorphism could be involved in the lower risk of coronary artery disease associated with the H1X447 genotype. PMID- 15356087 TI - The cholinergic system controls ghrelin release and ghrelin-induced growth hormone release in humans. AB - The stomach-derived peptide hormone ghrelin induces appetite and GH release. Several ghrelin actions are possibly mediated and modulated by the central cholinergic system. The aim of this study was to investigate the influence of the unspecific cholinergic antagonist atropine and the acetylcholine esterase inhibitor pyridostigmine, a cholinergic enhancer on ghrelin plasma concentrations and ghrelin-induced GH release. We investigated plasma ghrelin concentrations, ghrelin-induced GH release, and glucose and insulin concentrations after administration of atropine or pyridostigmine, and ghrelin (in two different doses, 0.25 and 1 microg/kg body weight), alone and in combination in a randomized, double-blind, placebo-controlled, crossover study design on 12 young, healthy male volunteers. Atropine alone significantly reduced fasting ghrelin levels by 25%, whereas under pyridostigmine alone ghrelin levels were unaltered. Ghrelin in combination with atropine induced significantly reduced GH concentrations compared with ghrelin administration alone for both ghrelin doses, whereas ghrelin-induced GH peak concentrations and areas under the curve were not enhanced by pyridostigmine treatment. These results suggest that, in humans, fasting ghrelin concentrations might be under cholinergic control and that the cholinergic system appears to modulate ghrelin-induced GH release. PMID- 15356088 TI - Affinity-enrichment of thyrotropin receptor autoantibodies from Graves' patients and normal individuals provides insight into their properties and possible origin from natural antibodies. AB - We used purified recombinant TSH receptor (TSHR) antigen prepared in mammalian cells to affinity-enrich TSHR autoantibodies from Graves' patients' IgG. Autoantibody enrichment, assayed by TSH binding inhibitory activity, was 20- to 1000-fold. Thyroid-stimulating antibody activity enrichment, although more difficult to quantitate, was comparable. TSHR-autoantibody approximate affinities for the holoreceptor assessed indirectly by TSH binding inhibition were 4-27 x 10(-9) m, an underestimate because 100% TSHR autoantibody purity was not attained. Consistent with previous data for serum, highly enriched TSHR autoantibodies in three of four patients showed lambda light chain bias. However, in contrast to expectations, antigen-enriched IgG was skewed primarily toward IgG2 and IgG3, subclasses associated with polysaccharides and microorganisms, respectively. Subclass depletion studies on antigen-enriched IgG indicated that TSHR autoantibodies were predominantly IgG1 and, surprisingly, IgG4. As controls, we affinity-enriched pooled IgG from normal individuals on TSHR antigen. This enriched IgG had detectable TSH binding inhibitory activity, although with lower specific activity than, and lacking the thyroid stimulatory activity of, Graves' IgG. Moreover, these natural IgG class autoantibodies largely recognized the same conformational variation in the TSHR N-terminal region as disease-associated TSHR autoantibodies. These studies suggest that TSHR autoantibodies may arise from natural autoantibodies, possibly by class switching from cross-reacting antibodies to microorganisms. PMID- 15356089 TI - Dual secretagogue drive of burst-like growth hormone secretion in postmenopausal compared with premenopausal women studied under an experimental estradiol clamp. AB - We show that in an experimentally enforced estradiol-predominant milieu, postmenopausal compared with premenopausal women maintain 1) decreased fasting GH and IGF-I concentrations, 2) reduced basal and pulsatile GH secretion, and 3) attenuated GH secretion after maximal stimulation by the paired secretagogues l arginine/GH-releasing peptide (GHRP)-2, l-arginine/GHRH, and GHRP-2/GHRH. These foregoing outcomes are selective, because menopausal status did not determine mean GH secretory-burst frequency or peptide-induced waveform shortening. Abdominal visceral fat mass predicted up to 25% of the variability in fasting and stimulated GH secretion in the combined cohorts under fixed systemic estradiol availability. Accordingly, as much as three-fourths of interindividual differences in burst-like GH secretion among healthy pre- and postmenopausal women arise from age-related mechanisms independently of short-term systemic estrogen availability and relative intraabdominal adiposity. PMID- 15356090 TI - 11beta-hydroxysteroid dehydrogenase type 1 activity in lean and obese males with type 2 diabetes mellitus. AB - Glucocorticoids play an important role in the pathogenesis of obesity and insulin resistance. Impaired conversion of cortisone (E) to cortisol (F) by the type 1 isoenzyme of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) in obesity may represent a protective mechanism preventing ongoing weight gain and glucose intolerance. We have studied glucocorticoid metabolism in 33 male subjects with type 2 diabetes mellitus [age, 44.2 +/- 13 yr; body mass index (BMI), 31.1 +/- 7.5 kg/m(2) (mean +/- sd)] and 38 normal controls (age, 41.4 +/- 14 yr; BMI, 38.2 +/- 12.8 kg/m(2)). Circulating F:E ratios were elevated in the diabetic group and correlated with serum cholesterol and homeostasis model assessment-S. There was no difference in 11beta-HSD1 activity between diabetic subjects and controls. In addition, 11beta-HSD1 activity was unaffected by BMI in diabetic subjects. However, in control subjects, increasing BMI was associated with a reduction in the urinary tetrahydrocortisol+5alpha-tetrahydrocortisol:tetrahydrocortisone ratio (P < 0.05) indicative of impaired 11beta-HSD1 activity. The degree of inhibition correlated tightly with visceral fat mass. Changes in 11beta-HSD1 activity could not be explained by circulating levels of adipocytokines. Impaired E to F metabolism in obesity may help preserve insulin sensitivity and prevent diabetes mellitus. Failure to down-regulate 11beta-HSD1 activity in patients with diabetes may potentiate dyslipidemia, insulin resistance, and obesity. Inhibition of 11beta-HSD1 may therefore represent a therapeutic strategy in patients with type 2 diabetes mellitus and obesity. PMID- 15356091 TI - Non-HDL cholesterol level is reliable to be an early predictor for vascular inflammation in type 2 diabetes mellitus. AB - Although LDL is the primary target for lipid-lowering therapy and non-HDL cholesterol is a secondary target in patients with elevated triglyceride (TG) levels, non-HDL cholesterol is still an early, reliable, and practical predictor for vascular inflammation. However, in comparison with LDL, further evidence for superiority in non-HDL cholesterol as the primary therapeutic target is required. A total of 189 type 2 diabetic patients (88 men and 101 women; mean age 58.4 +/- 14.0 years; duration of diabetes 9.8 +/- 4.2 years) who had not used anti inflammatory agents in the past two years were enrolled in this study. Levels of various lipid fractions and C-reactive protein (CRP) and the cholesterol retention fraction (CRF) were measured. Lipid levels and CRF in samples with CRP levels below or above different cutoffs were compared. Statistically significant differences were seen at all CRP cutoffs in the levels of TG, HDL, and non-HDL cholesterol and the CRF, but no differences were seen in total cholesterol and LDL cholesterol levels. CRP levels correlated with non-HDL cholesterol levels (r = 0.16, P = 0.0236) and the CRF (r = 0.18, P = 0.14), but not with levels of HDL or TG. Besides, non-HDL levels showed a marked correlation with CRF (r = 0.68, P < 0.0001). On the basis of CRP levels, non-HDL levels are reliable in predicting vascular inflammation, and CRF could be another important predictor for cardiovascular events. Our results suggest that the emphasis placed on non-HDL cholesterol should be reevaluated in comparison with that placed on LDL cholesterol. PMID- 15356092 TI - N-octanoylated ghrelin levels in cord and neonatal blood. PMID- 15356093 TI - Should serum calcitonin be routinely measured in patients with thyroid nodules- will the law answer before endocrinologists do? PMID- 15356094 TI - Low vitamin B12 and bone loss: a role for folate deficiency. PMID- 15356095 TI - Two cases of thyroid carcinoma that were not stimulated by recombinant human thyrotropin. PMID- 15356096 TI - Association of glutathione peroxidase activity with insulin resistance and dietary fat intake during normal pregnancy. PMID- 15356097 TI - A novel epitope of N-CAM defines precursors of human adherent NK cells. AB - Activated, adherent natural killer (A-NK) cells represent a distinct subpopulation of interleukin (IL)-2-stimulated NK cells, which are selectively endowed with the increased expression of integrins and ability to adhere to solid surfaces, migrate into, infiltrate, and destroy cancerous tissues. The present study defines the phenotype and functions of precursors of A-NK (pre-A-NK) cells in humans. Peripheral blood pre-A-NK cells, in contrast to the rest of NK cells, express a novel epitope of CD56 neuronal cell adhesion molecule, termed ANK-1, and increased cell-surface levels of integrins. Pre-A-NK cells also express low levels of CD56 and CD161, and some express CD162 receptor, do not express CD25 or activation markers, and are effective mediators of NK cytotoxicity. Thus, pre-A NK cells are generally similar to CD56(dim) NK cells. However, pre-A-NK cells differ from the main NK cell subpopulation by having a lower expression level of CD16 and a lower ability to mediate redirected antibody-dependent, cell-mediated cytotoxicity. More importantly, pre-A-NK cells are preferentially endowed with the ability to rapidly respond to IL-2 by integrin-mediated adherence to endothelial cells, extracellular matrix, and plastic. This early, specific response of pre-A-NK cells to IL-2 is followed by their activation, vigorous proliferation, and differentiation into phenotypically and functionally similar A NK cells. Pre-A-NK cells represent only approximately 26% of peripheral blood NK cells but encompass the majority of NK cells in normal and cancerous, solid tissues. We conclude that pre-A-NK cells represent a distinct subset of resting, mature NK cells with the characteristics indicative of their ability to migrate and reside in solid tissues. PMID- 15356098 TI - Rat NKp46 activates natural killer cell cytotoxicity and is associated with FcepsilonRIgamma and CD3zeta. AB - NKp46 has been identified in the human, rat, mouse, monkey, and cattle. We have generated a monoclonal antibody, WEN23, against rat NKp46. By flow cytometry, NKp46 is expressed by all natural killer (NK) cells but not by T cells, B cells, granulocytes, monocytes, dendritic cells, or macrophages. Thus, NKp46/WEN23 is the first NK cell-specific marker in the rat. In a redirected lysis assay, preincubation of the effector cells with WEN23 augmented lysis of the Fc receptor (FcR)+ murine tumor target cells, indicating that NKp46 is an activating NK cell receptor. Moreover, preincubation of the effector cells with WEN23 F(ab')2 fragments reduced killing of target cells, confirming the activating function of NKp46 and indicating that the mouse tumor target cells express a ligand for rat NKp46. Lysis of FcR- mouse and human tumor target cells was reduced after incubation of effector cells with WEN23, suggesting that rat NKp46 recognizes a ligand that is conserved between primates and rodents. By Western blot and immunoprecipitation using WEN23, NKp46 is expressed as a monomer of approximately 47 kDa in interleukin-2-activated NK cells. The immunoreceptor tyrosine-based activation motif bearing adaptor proteins CD3zeta and the gamma chain of FcRI for IgE (FcepsilonRIgamma) with NKp46 from lysates of NK cells, indicating that rat NKp46 activates NK cell cytotoxicity by similar pathways as CD16. PMID- 15356099 TI - Granulocyte chemotactic protein-2 mediates adaptive immunity in part through IL 8Rbeta interactions. AB - Chemokines constitute a large family of structurally related proteins that play a role in leukocyte migration and differentiation. Indeed, the early expression of human CXC chemokine receptor 1 (hCXCR1) and hCXCR2 [homologous to mouse interleukin (IL)-8Rbeta] ligands by the epithelium is a hallmark of the mucosal host defense. Mice lack IL-8; however, granulocyte chemotactic protein-2 (GCP 2)/lipopolysaccharide-induced CXC chemokine, a murine homologue of human GCP-2, has 32% and 61% sequence identity to human IL-8 and GCP-2, respectively, and binds hCXCR1, hCXCR2, and mouse IL-8Rbeta. To better understand the role of GCP-2 in adaptive immunity and as a nasal adjuvant, we characterized the exogenous effects of this CXC chemokine on cellular and humoral mucosal immune responses. GCP-2 significantly enhanced serum immunoglobulin G (IgG) and mucosal IgA antibodies through increased cytokine secretion by CD4+ T cells. These alterations in humoral and cellular responses were preceded by an increase in the number of B cells in the nasal tract, a decrease in the number of CD4+ T cells in the nasal tract as well as cervical lymph nodes, and an increase in the number of neutrophils in the nasal tract 12 h after GCP-2 immunization. This chemokine also modulated CD28 expression by CD4+ T cells during CD3epsilon stimulation of wild type mice. GCP-2 increased CD80 and CD86 expression on B cells during in vitro stimulation in a dose-dependent manner. In contrast, cytokine and costimulatory molecule enhancement by GCP-2 was not induced by lymphocytes from IL-8Rbeta-/- mice, suggesting that GCP-2 modulates cellular immunity in part through IL-8Rbeta interactions. PMID- 15356100 TI - Thy-1: more than a mouse pan-T cell marker. AB - Thy-1 (CD90) is a small GPI-anchored protein that is particularly abundant on the surface of mouse thymocytes and peripheral T cells. T cell proliferation and cytokine synthesis in response to Thy-1 cross-linking by specific mAb suggests a role for Thy-1 in mouse T lymphocyte activation. However, a physiological ligand or counterreceptor for murine Thy-1 in the lymphoid compartment has not yet been identified. Thy-1 cross-linking, in the context of strong costimulatory signaling through CD28, results in an activating signal that can at least partially substitute for TCR signaling during mouse T cell activation. Remarkably, Thy-1 cross-linking also results in the potent costimulation of T cells activated through the TCR. This novel dual signaling capacity suggests a possible role for Thy-1 in the maintenance of T cell homeostasis in the absence of TCR triggering, as well as potentiating Ag-induced T cell responses. PMID- 15356101 TI - Cutting edge: direct interaction of TLR4 with NAD(P)H oxidase 4 isozyme is essential for lipopolysaccharide-induced production of reactive oxygen species and activation of NF-kappa B. AB - LPS, the primary constituent of the outer membrane of Gram-negative bacteria, is recognized by TLR4. Binding of TLR4 to LPS triggers various cell signaling pathways including NF-kappaB activation and reactive oxygen species (ROS) production. In this study, we present the data that LPS-induced ROS generation and NF-kappaB activation are mediated by a direct interaction of TLR4 with (NAD(P)H oxidase 4 (Nox) 4), a protein related to gp91phox (Nox2) of phagocytic cells, in HEK293T cells. Yeast two hybrid and GST pull-down assays indicated that the COOH-terminal region of Nox4 interacted with the cytoplasmic tail of TLR4. Knockdown of Nox4 by transfection of small interference RNA specific to the Nox4 isozyme in HEK293T cells expressing TLR4 along with MD2 and CD14 resulted in inhibition of LPS-induced ROS generation and NF-kappaB activation. Taken together, these results indicate that direct interaction of TLR4 with Nox4 is involved in LPS-mediated ROS generation and NF-kappaB activation. PMID- 15356102 TI - Cutting edge: murine dendritic cells require IL-15R alpha to prime NK cells. AB - NK cells protect hosts against viral pathogens and transformed cells, and dendritic cells (DCs) play important roles in activating NK cells. We now find that murine IL-15Ralpha-deficient DCs fail to support NK cell cytolytic activity and elaboration of IFN-gamma, despite the fact that these DCs express normal levels of costimulatory molecules and IL-12. By contrast, IL-15Ralpha expression on NK cells is entirely dispensable for their activation by DCs. In addition, blockade with anti-IL-15Ralpha and anti-IL-2Rbeta but not anti-IL-2Ralpha specific Abs prevents NK cell activation by wild-type DCs. Finally, presentation of IL-15 by purified IL-15Ralpha/Fc in trans synergizes with IL-12 to support NK cell priming. These findings suggest that murine DCs require IL-15Ralpha to present IL-15 in trans to NK cells during NK cell priming. PMID- 15356103 TI - Cutting edge: serotonin is a chemotactic factor for eosinophils and functions additively with eotaxin. AB - Elevated levels of serotonin (5-hydroxytryptamine, 5-HT) are observed in the serum of asthmatics. Herein, we demonstrate that 5-HT functions independently as an eosinophil chemoattractant that acts additively with eotaxin. 5-HT2A receptor antagonists (including MDL-100907 and cyproheptadine (CYP)) were found to inhibit 5-HT-induced, but not eotaxin-induced migration. Intravital microscopy studies revealed that eosinophils roll in response to 5-HT in venules under conditions of physiological shear stress, which could be blocked by pretreating eosinophils with CYP. OVA-induced pulmonary eosinophilia in wild-type mice was significantly inhibited using CYP alone and maximally in combination with a CCR3 receptor antagonist. Interestingly, OVA-induced pulmonary eosinophilia in eotaxin-knockout (Eot-/-) mice was inhibited by treatment with the 5-HT2A but not CCR3 receptor antagonist. These results suggest that 5-HT is a potent eosinophil-active chemoattractant that can function additively with eotaxin and a dual CCR3/5-HT2A receptor antagonist may be more effective in blocking allergen-induced eosinophil recruitment. PMID- 15356104 TI - Stromelysin-2 (matrix metalloproteinase 10) is inducible in lymphoma cells and accelerates the growth of lymphoid tumors in vivo. AB - Matrix metalloproteinase (MMP) 10 (stromelysin-2) is known to degrade various components of the extracellular matrix; however, the signals that regulate its expression and its role in lymphoma growth remain unknown. In the present work, we report the up-regulated expression of MMP10 in T lymphoma cells following contact with endothelial cells. The induction of MMP10 was found to be dependent on the specific interaction between LFA-1 and ICAM-1, which play a central role in regulating the expression of genes involved in the rate-limiting steps of lymphoma development. MMP10, but not MMP3 (stromelysin-1), was also up-regulated in human B lymphoma cells following exposure to IL-4, IL-6, and IL-13, but not to IL-1. To gain further insight into the role of MMP10 in lymphoma development, we generated lymphoma cell lines constitutively expressing high levels of MMP10 and studied these cells for their ability to form thymic lymphoma in vivo. Mice injected with lymphoma cells constitutively expressing MMP10 developed thymic lymphoma more rapidly than those injected with control lymphoma cells. These results provide the first in vivo evidence that overexpression of MMP10 promotes tumor development, and indicate that MMP10 induction is an important pathway activated not only upon ICAM-1/LFA-1-mediated intercellular contact, but also following activation of tumor cells with inflammatory cytokines. PMID- 15356105 TI - Adoptive transfer of paternal antigen-hyporesponsive T cells induces maternal tolerance to the allogeneic fetus in abortion-prone matings. AB - The embryo expresses paternal Ags foreign to the mother and therefore has been viewed as an allograft. It has been shown that anergic T cells generated by blocking of the CD28/B7 costimulatory pathway with anti B7-1 and anti B7-2 mAbs can be transferred as suppresser cells to prevent allograft rejection. Little is known, however, about the in vivo function of anti-B7-treated T cells after their transfer into abortion-prone mice in the maintenance of materno-fetal tolerance. In the present study, abortion-prone CBA/J females mated with DBA/2 males were administered anti-B7-1 and anti-B7-2 mAbs on day 4 of gestation (murine implantation window). The anti-B7-treated T cells subsequently were adoptively transferred into abortion-prone CBA/J mice. We demonstrated that costimulation blockade with anti-B7 mAbs at the time of implantation resulted in altered allogeneic T cell response and overcame increased maternal rejection to the fetus in the CBA/JxDBA/2 system. The transferred anti-B7-treated T cells appeared to be regulatory, decreasing responsiveness and generating clonal deviation in maternal recipient T cells. The transferred CFSE-labeled T cells were found to reside in the spleen and uterine draining lymph nodes, and a few were localized to the materno-fetal interface of the maternal recipient. Our findings suggest that the anti-B7-treated T cells not only function as potent suppresser cells, but also exert an immunoregulatory effect on the maternal recipient T cells, which cosuppresses maternal rejection to the fetus. This procedure might be considered potentially useful for fetal survival when used as an immunotherapy for human recurrent spontaneous abortion. PMID- 15356106 TI - Kinetics of in vivo elimination of suicide gene-expressing T cells affects engraftment, graft-versus-host disease, and graft-versus-leukemia after allogeneic bone marrow transplantation. AB - Suicide gene therapy is one approach being evaluated for the control of graft-vs host disease (GVHD) after allogeneic bone marrow transplantation (BMT). We recently constructed a novel chimeric suicide gene in which the entire coding region of HSV thymidine kinase (HSV-tk) was fused in-frame to the extracellular and transmembrane domains of human CD34 (DeltaCD34-tk). DeltaCD34-tk is an attractive candidate as a suicide gene in man because of the ensured expression of HSV-tk in all selected cells and the ability to rapidly and efficiently purify gene-modified cells using clinically approved CD34 immunoselection techniques. In this study we assessed the efficacy of the DeltaCD34-tk suicide gene in the absence of extended ex vivo manipulation by generating transgenic animals that express DeltaCD34-tk in the peripheral and thymic T cell compartments using the CD2 locus control region. We found that DeltaCD34-tk-expressing T cells could be purified to near homogeneity by CD34 immunoselection and selectively eliminated ex vivo and in vivo when exposed to low concentrations of GCV. The optimal time to administer GCV after allogeneic BMT with DeltaCD34-tk-expressing transgenic T cells was dependent on the intensity of the conditioning regimen, the leukemic status of the recipient, and the dose and timing of T cell infusion. Importantly, we used a controlled graft-vs-host reaction to promote alloengraftment in sublethally irradiated mice and provide a graft-vs-leukemia effect in recipients administered a delayed infusion of DeltaCD34-tk-expressing T cells. This murine model demonstrates the potential usefulness of DeltaCD34-tk-expressing T cells to control GVHD, promote alloengraftment, and provide a graft-vs-leukemia effect in man. PMID- 15356107 TI - B7-2 (CD86) controls the priming of autoreactive CD4 T cell response against pancreatic islets. AB - The B7-1/2-CD28 system provides the critical signal for the generation of an efficient T cell response. We investigated the role played by B7-2 in influencing pathogenic autoimmunity from islet-reactive CD4 T cells in B7-2 knockout (KO) NOD mice which are protected from type 1 diabetes. B7-2 deficiency caused a profound diminishment in the generation of spontaneously activated CD4 T cells and islet specific CD4 T cell expansion. B7-2 does not impact the effector phase of the autoimmune response as adoptive transfer of islet Ag-specific BDC2.5 splenocytes stimulated in vitro could easily induce disease in B7-2KO mice. CD4 T cells showed some hallmarks of hyporesponsiveness because TCR/CD28-mediated stimulation led to defective activation and failure to induce disease in NODscid recipients. Furthermore, CD4 T cells exhibited enhanced death in the absence of B7-2. Interestingly, we found that B7-2 is required to achieve normal levels of CD4+CD25+CD62L+ T regulatory cells because a significant reduction of these T regulatory cells was observed in the thymus but not in the peripheral compartments of B7-2KO mice. In addition, our adoptive transfer experiments did not reveal either pathogenic or regulatory potential associated with the B7-2KO splenocytes. Finally, we found that the lack of B7-2 did not induce a compensatory increase in the B7-1 signal on APC in the PLN compartment. Taken together these results clearly indicate that B7-2 plays a critical role in priming islet-reactive CD4 T cells, suggesting a simplified, two-cell model for the impact of this costimulatory molecule in autoimmunity against islets. PMID- 15356108 TI - Dynamic redistribution of the activating 2B4/SAP complex at the cytotoxic NK cell immune synapse. AB - The 2B4 molecule (CD244) has been described as a coreceptor in human NK cell activation. However, the behavior of 2B4 during the cytotoxic NK cell immune synapse (NK-IS) formation remains undetermined. In this study, we demonstrate the redistribution of 2B4 and the signaling adaptor molecule, signaling lymphocyte activation molecule-associated protein (SAP), to the cytotoxic NK-IS upon formation of conjugates between resting NK cells and EBV-infected 721.221 human cells. Confocal microscopy showed that 2B4 localized at the central supramolecular activation cluster, surrounded by a peripheral supramolecular activation cluster containing talin within NK cell and ICAM-1 on target cells. Videomicroscopy studies with 2B4-GFP-transfected NK cells revealed that 2B4 redistributed to cytotoxic NK-IS as soon as the cell contact occurred. Simultaneously, a SAP-GFP also clustered at the contact site, where it remained during the interaction period. The 2B4 molecular clusters remained bound to the target cell even after NK cell detachment. These results underscore the function of 2B4 as an adhesion molecule and suggest a relevant role in the initial binding, scanning of target cells, and formation of cytotoxic NK-IS. Finally, these findings are indicative of an important role of the activating 2B4/signaling lymphocyte activation molecule-associated protein complex during the recognition of EBV-infected cells. PMID- 15356109 TI - Immunological synapse formation licenses CD40-CD40L accumulations at T-APC contact sites. AB - The maintenance of tolerance is likely to rely on the ability of a T cell to polarize surface molecules providing "help" to only specific APCs. The formation of a mature immunological synapse leads to concentration of the TCR at the APC interface. In this study, we show that the CD40-CD154 receptor-ligand pair is also highly concentrated into a central region of the synapse on mouse lymphocytes only after the formation of the TCR/CD3 c-SMAC. Concentration of this ligand was strictly dependent on TCR recognition, the binding of ICAM-1 to T cell integrins and the presence of an intact cytoskeleton in the T cells. This may provide a novel explanation for the specificity of T cell help directing the help signal to the site of Ag receptor signal. It may also serve as a site for these molecular aggregates to coassociate and/or internalize alongside other signaling receptors. PMID- 15356110 TI - LFA-1 contributes an early signal for NK cell cytotoxicity. AB - Cytotoxicity of human NK cells is activated by receptors that bind ligands on target cells, but the relative contribution of the many different activating and inhibitory NK cell receptors is difficult to assess. In this study, we describe an experimental system that circumvents some of the difficulties. Adhesion through beta2 integrin LFA-1 is a common requirement of CTLs and NK cells for efficient lysis of target cells. However, the contribution of LFA-1 to activation signals for NK cell cytotoxicity, besides its role in adhesion, is unclear. The role of LFA-1 was evaluated by exposing NK cells to human ICAM-1 that was either expressed on a Drosophila insect cell line, or directly coupled to beads. Expression of ICAM-1 on insect cells was sufficient to induce lysis by NK cells through LFA-1. Coexpression of peptide-loaded HLA-C with ICAM-1 on insect cells blocked the LFA-1-dependent cytotoxicity of NK cells that expressed HLA-C specific inhibitory receptors. Polarization of cytotoxic granules in NK cells toward ICAM-1- and ICAM-2-coated beads showed that engagement of LFA-1 alone is sufficient to initiate activation signals in NK cells. Thus, in contrast to T cells, in which even adhesion through LFA-1 is dependent on signals from other receptors, NK cells receive early activation signals directly through LFA-1. PMID- 15356111 TI - Fas-Fas ligand interactions are essential for the binding to and killing of activated macrophages by gamma delta T cells. AB - Gammadelta T cells have a direct role in resolving the host immune response to infection by eliminating populations of activated macrophages. Macrophage reactivity resides within the Vgamma1/Vdelta6.3 subset of gammadelta T cells, which have the ability to kill activated macrophages following infection with Listeria monocytogenes (Lm). However, it is not known how gammadelta T cell macrophage cytocidal activity is regulated, or what effector mechanisms gammadelta T cells use to kill activated macrophages. Using a macrophage-T cell coculture system in which peritoneal macrophages from naive or Lm-infected TCRdelta-/- mice were incubated with splenocytes from wild-type and Fas ligand (FasL)-deficient mice (gld), the ability of Vgamma1 T cells to bind macrophages was shown to be dependent upon Fas-FasL interactions. Combinations of anti-TCR and FasL Abs completely abolished binding to and killing of activated macrophages by Vgamma1 T cells. In addition, confocal microscopy showed that Fas and the TCR colocalized on Vgamma1 T cells at points of contact with macrophages. Collectively, these studies identify an accessory or coreceptor-like function for Fas-FasL that is essential for the interaction of Vgamma1 T cells with activated macrophages and their elimination during the resolution stage of pathogen-induced immune responses. PMID- 15356112 TI - CC chemokine ligands 25 and 28 play essential roles in intestinal extravasation of IgA antibody-secreting cells. AB - CCL25 (also known as thymus-expressed chemokine) and CCL28 (also known as mucosae associated epithelial chemokine) play important roles in mucosal immunity by recruiting IgA Ab-secreting cells (ASCs) into mucosal lamina propria. However, their exact roles in vivo still remain to be defined. In this study, we first demonstrated in mice that IgA ASCs in small intestine expressed CCR9, CCR10, and CXCR4 on the cell surface and migrated to their respective ligands CCL25, CCL28, and CXCL12 (also known as stromal cell-derived factor 1), whereas IgA ASCs in colon mainly expressed CCR10 and CXCR4 and migrated to CCL28 and CXCL12. Reciprocally, the epithelial cells of small intestine were immunologically positive for CCL25 and CCL28, whereas those of colon were positive for CCL28 and CXCL12. Furthermore, the venular endothelial cells in small intestine were positive for CCL25 and CCL28, whereas those in colon were positive for CCL28, suggesting their direct roles in extravasation of IgA ASCs. Consistently, in mice orally immunized with cholera toxin (CT), anti-CCL25 suppressed homing of CT specific IgA ASCs into small intestine, whereas anti-CCL28 suppressed homing of CT-specific IgA ASCs into both small intestine and colon. Reciprocally, CT specific ASCs and IgA titers in the blood were increased in mice treated with anti-CCL25 or anti-CCL28. Anti-CXCL12 had no such effects. Finally, both CCL25 and CCL28 were capable of enhancing alpha4 integrin-dependent adhesion of IgA ASCs to mucosal addressin cell adhesion molecule-1 and VCAM-1. Collectively, CCL25 and CCL28 play essential roles in intestinal homing of IgA ASCs primarily by mediating their extravasation into intestinal lamina propria. PMID- 15356113 TI - Nitric oxide-dependent mitochondrial biogenesis generates Ca2+ signaling profile of lupus T cells. AB - Abnormal T cell activation and cell death underlie the pathology of systemic lupus erythematosus. Although mitochondrial hyperpolarization (MHP) represents an early and reversible checkpoint of T cell activation and apoptosis, lupus T cells exhibit persistent MHP. NO has recently been recognized as a key signal of mitochondrial biogenesis and mediator of MHP in human T lymphocytes. In this study, we show that persistent MHP was associated with increased mitochondrial mass (+47.7 +/- 2.8%; p = 0.00017) and increased mitochondrial (+21.8 +/- 4.1%; p = 0.016) and cytoplasmic Ca2+ content in T cells from 19 systemic lupus erythematosus patients with respect to 11 control donors (+38.0 +/- 6.4%; p = 0.0023). Electron microscopy revealed that lupus lymphocytes contained 8.76 +/- 1.0 mitochondria, while control donors contained 3.18 +/- 0.28 mitochondria per cell (p = 0.0009). Increased mitochondrial mass in T cells was associated with 2.08 +/- 0.09-fold enhanced NO production by lupus monocytes (p = 0.0023). Activation of T cells through the TCR initiates a biphasic elevation in cytosolic free Ca2+ concentration, a rapid initial peak observed within minutes, and a plateau phase lasting up to 48 h. In response to CD3/CD28 costimulation, rapid Ca2+ fluxing was enhanced while the plateau phase was diminished in lupus T cells. NO-induced mitochondrial biogenesis in normal T cells enhanced the rapid phase and reduced the plateau of Ca2+ influx upon CD3/CD28 costimulation, thus mimicking the Ca2+ signaling profile of lupus T cells. Mitochondria constitute major Ca2+ stores and NO-dependent mitochondrial biogenesis may account for altered Ca2+ handling by lupus T cells. PMID- 15356114 TI - CD59a is the primary regulator of membrane attack complex assembly in the mouse. AB - Gene-deleted mice have provided a potent tool in efforts to understand the roles of complement and complement-regulating proteins in vivo. In particular, mice deficient in the membrane regulators complement receptor 1-related gene/protein y, decay-accelerating factor, or CD59 have demonstrated homeostatic relevance and backcrossing between the strains has revealed cooperativity in regulation. In mouse, genes encoding decay-accelerating factor and CD59 have been duplicated and show differential expression in tissues, complicating interpretation and extrapolation of findings to man. The first described form of CD59, CD59a, is broadly distributed and deletion of the cd59a gene causes a mild hemolytic phenotype with increased susceptibility in complement-mediated disease models. The distribution of the second form, CD59b, was originally described as testis specific, but later by some as widespread. Deletion of the cd59b gene caused a severe hemolytic and thrombotic phenotype. To apply data from these mouse models to man it is essential to know the relative distribution and functional roles of these two forms of CD59. We have generated new specific reagents and used them in sensitive quantitative analyses to comprehensively characterize expression of mRNA and protein and functional roles of CD59a and CD59b in wild-type (wt) and CD59a-negative mice. cd59b mRNA was detected only in testis and, at very low levels, in bone marrow. CD59b protein was present on mature spermatozoa and precursors and, in trace amounts, erythrocytes. Erythrocyte CD59b did not inhibit complement lysis except when CD59a was absent or blocked. These data confirm that CD59a is the primary regulator of complement membrane attack in mouse. PMID- 15356115 TI - Quantitative and qualitative differences in the in vivo response of NKT cells to distinct alpha- and beta-anomeric glycolipids. AB - NKT cells represent a unique subset of immunoregulatory T cells that recognize glycolipid Ags presented by the MHC class I-like molecule CD1d. Because of their immunoregulatory properties, NKT cells are attractive targets for the development of immunotherapies. The prototypical NKT cell ligand alpha-galactosylceramide (alpha-GalCer), originally isolated from a marine sponge, has potent immunomodulatory activities in mice, demonstrating therapeutic efficacy against metastatic tumors, infections, and autoimmune diseases, but also has a number of adverse side effects. In vivo administration of alpha-GalCer to mice results in the rapid activation of NKT cells, which is characterized by cytokine secretion, surface receptor down-regulation, expansion, and secondary activation of a variety of innate and adaptive immune system cells. In this study, we have evaluated the in vivo immune response of mice to a set of structural analogues of alpha-GalCer. Our results show that, contrary to current thinking, beta-anomeric GalCer can induce CD1d-dependent biological activities in mice, albeit at lower potency than alpha-anomeric GalCer. In addition, we show that the response of NKT cells to distinct GalCer differs not only quantitatively, but also qualitatively. These findings indicate that NKT cells can fine-tune their immune responses to distinct glycolipid Ags in vivo, a property that may be exploited for the development of effective and safe NKT cell-based immunotherapies. PMID- 15356116 TI - Differential requirement for tapasin in the presentation of leader- and insulin derived peptide antigens to Qa-1b-restricted CTLs. AB - The loading of MHC class I molecules with peptides involves a variety of accessory proteins, including TAP-associated glycoprotein (tapasin), which tethers empty MHC class I molecules to the TAP peptide transporter. We have evaluated the role of tapasin for the assembly of peptides with the class Ib molecule Qa-1b. In normal cells, Qa-1b is predominantly bound by a peptide, the Qa-1 determinant modifier (Qdm), derived from the signal sequence of class Ia molecules. Our results show that tapasin links Qa-1b to the TAP peptide transporter, and that tapasin facilitates the delivery of Qa-1b molecules to the cell surface. Tapasin was also required for the presentation of endogenous Qdm peptides to Qdm-specific, Qa-1b-restricted CTLs. In sharp contrast, tapasin expression was dispensable for the presentation of an insulin peptide to insulin specific, Qa-1b-restricted CTL isolated from TCR transgenic mice. However, tapasin deficiency significantly impaired the positive selection of these insulin specific, Qa-1b-restricted transgenic CD8+ T cells. These findings reveal that tapasin plays a differential role in the loading of Qdm and insulin peptides onto Qa-1b molecules, and that tapasin is dispensable for retention of empty Qa-1b molecules in the endoplasmic reticulum, and are consistent with the proposed peptide-editing function of tapasin. PMID- 15356117 TI - Cross-talk between activated human NK cells and CD4+ T cells via OX40-OX40 ligand interactions. AB - It is important to understand which molecules are relevant for linking innate and adaptive immune cells. In this study, we show that OX40 ligand is selectively induced on IL-2, IL-12, or IL-15-activated human NK cells following stimulation through NKG2D, the low affinity receptor for IgG (CD16) or killer cell Ig-like receptor 2DS2. CD16-activated NK cells costimulate TCR-induced proliferation, and IFN-gamma produced by autologous CD4+ T cells and this process is dependent upon expression of OX40 ligand and B7 by the activated NK cells. These findings suggest a novel and unexpected link between the natural and specific immune responses, providing direct evidence for cross-talk between human CD4+ T cells and NK receptor-activated NK cells. PMID- 15356118 TI - Stimulatory killer Ig-like receptors modulate T cell activation through DAP12 dependent and DAP12-independent mechanisms. AB - Stimulatory killer Ig-like receptors (KIRs) are expressed by various lymphocytes, including NK cells and subsets of T cells. In NK cells, KIRs associate with the adapter molecule KARAP/DAP12, which confers the ability to function as an independent activation unit. The function of KIRs and killer cell activating receptor-associated protein (KARAP)/DAP12 in T cells is unclear. By flow cytometry, we demonstrated that CD4+CD28null T cells heterogeneously express KIRs and/or KARAP/DAP12. In clones that lacked expression of KARAP/DAP12, the stimulatory KIR KIR2DS2 signaled through the JNK pathway, but did not activate the ERK pathway. However, in the presence of KARAP/DAP12, stimulation through KIR2DS2 led to phosphorylation of both JNK and ERK. Transfection experiments confirmed that KIR2DS2-mediated ERK phosphorylation was dependent on KARAP/DAP12. The differential signaling of KIR2DS2 through association with alternative adapter molecules resulted in differential regulation of cellular activity. In clones that lacked expression of KARAP/DAP12, stimulation of KIR2DS2 did not induce cytotoxicity. However, KIR2DS2 did augment suboptimal TCR stimulation, leading to enhanced IFN-gamma production. In clones that expressed KARAP/DAP12, KIR2DS2 directly activated both cytotoxicity and IFN-gamma production without the need for TCR-derived signals. The function of stimulatory KIRs in T cells is determined by the expression of the appropriate adapter molecule. Expression of KARAP/DAP12 is sufficient to convert a costimulatory KIR into a stimulatory molecule. These differing functions mediated by alternative signaling pathways have implications for the pathogenesis of diseases such as rheumatoid arthritis and acute coronary syndromes, in which aberrant expression of KIRs on T cells is frequently observed. PMID- 15356119 TI - The critical role of residues 43R and 44Q of carcinoembryonic antigen cell adhesion molecules-1 in the protection from killing by human NK cells. AB - The multifunctional carcinoembryonic Ag cell adhesion molecule (CEACAM)1 protein has recently become the focus of intense immunological research. We have previously shown that the CEACAM1 homophilic interactions inhibit the killing activity of NK cells. This novel inhibitory mechanism plays a key role in melanoma immune evasion, inhibition of decidual immune response, and controlling NK autoreactivity in TAP2-deficient patients. These roles are mediated mainly by homophilic interactions, which are mediated through the N-domain of the CEACAM1. The N-domain of the various members of the CEACAM family shares a high degree of similarity. However, it is still unclear which of the CEACAM family members is able to interact with CEACAM1 and what are the amino acid residues that control this interaction. In this study we demonstrate that CEACAM1 interacts with CEACAM5, but not with CEACAM6. Importantly, we provide the molecular basis for CEACAM1 recognition of various CEACAM family members. Sequence alignment reveals a dichotomy among the CEACAM family members: both CEACAM1 and CEACAM5 contain the R and Q residues in positions 43 and 44, respectively, whereas CEACAM3 and CEACAM6 contain the S and L residues, respectively. Mutational analysis revealed that both 43R and 44Q residues are necessary for CEACAM1 interactions. Implications for differential expression of CEACAM family members in tumors are discussed. PMID- 15356120 TI - A dual role of IFN-alpha in the balance between proliferation and death of human CD4+ T lymphocytes during primary response. AB - Type I IFNs (IFN-alphabeta) enhance immune responses, notably T cell-mediated responses, in part by promoting the functional activities of dendritic cells. In this study, we analyzed the direct impact of IFN-alpha on proliferative and apoptotic signals upon in vitro activation of human naive CD4+ T lymphocytes. We demonstrate that IFN-alpha protects T cells from the intrinsic mitochondrial dependent apoptosis early upon TCR/CD28 activation. IFN-alpha acts by delaying entry of cells into the G1 phase of the cell cycle, as well as by increasing Bcl 2 and limiting Bax activation. Later, upon activation, T cells that were exposed to IFN-alpha showed increased levels of surface Fas associated with partially processed caspase-8, a key component of the extrinsic apoptotic pathway. Caspase 8 processing was augmented furthermore by Fas ligation. Overall, these findings support a model whereby IFN-alpha favors an enhanced clonal expansion, yet it sensitizes cells to the Ag-induced cell death occurring at the end of an immune response. These observations point to a complex role of type I IFN in regulating the magnitude of proliferation and survival of naive CD4+ T cells during primary response and underline how crucial could be the timing of exposure to this cytokine. PMID- 15356121 TI - Murine plasmacytoid dendritic cells initiate the immunosuppressive pathway of tryptophan catabolism in response to CD200 receptor engagement. AB - In this study, using a soluble CD200-Ig fusion protein, we provide evidence that murine dendritic cells (DCs) possess a functional CD200R, whose engagement results in the reinforcement or appearance of immunosuppressive properties in these cells. In particular, the plasmacytoid subset (CD11c+B220+120G8+) of splenic DCs (pDCs) is induced by CD200-Ig to express the enzyme IDO, which initiates the tolerogenic pathway of tryptophan catabolism. As a result, pDCs are capable of suppressing Ag-specific responses in vivo when transferred into recipient hosts after treatment with CD200-Ig. IDO induction in pDCs through CD200R engagement requires type I IFNR signaling. Although the release of IFN alpha may contribute to the full expression of CD200-Ig activity, autocrine IFN alpha is unlikely to mediate alone the effects of CD200R engagement. These data prospect novel functions for both pDCs and the CD200-CD200R pair in the mouse. At the same time, these data underscore the possible unifying role of the IDO mechanism in immune tolerance. PMID- 15356122 TI - Intratumoral CC chemokine ligand 5 overexpression delays tumor growth and increases tumor cell infiltration. AB - Chemokines participate in the antitumor immune response by regulating the movement and positioning of lymphocytes as well as effector functions and may thus be candidates for use in antitumor therapy. To test whether CCL5, a chemokine involved in the recruitment of a wide spectrum of immunocompetent cells, can control tumor growth, we forced its expression at mouse tumor sites. Tumor growth was reduced in mice with s.c. syngeneic CCL5-EL-4 compared with EL-4 injected mice, whereas both reduced tumor growth and incidence were observed in mice with OVA-expressing EG-7 transfected with CCL5 compared with EG-7-injected mice. Significant antitumor effects were observed soon after intratumoral injection of DNA plasmid coding for chimeric CCL5-Ig. Importantly, quantitative RT-PCR assays showed that the amount of CCL5 expression at the tumor site determined the effectiveness of the antitumor response, which was associated with infiltration of increased numbers of NK, CD4, and CD8 cells at the tumor site. This effect was lost in mice deficient for T/B lymphocytes (RAG-2 knockout) or for CCR5 (CCR5 knockout). Together, these data demonstrate the antitumor activity of intratumoral CCL5 overexpression, due to its recruitment of immunocompetent cells, and the potential usefulness of chimeric CCL5-Ig DNA as an agent in cancer therapy. PMID- 15356123 TI - Discrete event modeling of CD4+ memory T cell generation. AB - Studies of memory T cell differentiation are hampered by a lack of quantitative models to test hypotheses in silico before in vivo experimentation. We created a stochastic computer model of CD4+ memory T cell generation that can simulate and track 10(1)-10(8) individual lymphocytes over time. Parameters for the model were derived from experimental data using naive human CD4+ T cells stimulated in vitro. Using discrete event computer simulation, we identified two key variables that heavily influence effector burst size and the persistent memory pool size: the cell cycle dependent probability of apoptosis, and the postactivation mitosis at which memory T cells emerge. Multiple simulations were performed and varying critical parameters permitted estimates of how sensitive the model was to changes in all of the model parameters. We then compared two hypotheses of CD4+ memory T cell generation: maturation from activated naive to effector to memory cells (model I) vs direct progression from activated naive to memory cells (model II). We find that direct progression of naive to memory T cells does not explain published measurements of the memory cell mass unless postactivation expansion of the memory cell cohort occurs. We conclude that current models suggesting direct progression of activated naive cells to the persistent memory phenotype (model II) do not account for the experimentally measured size of the postactivation CD4+, Ag-specific, memory T cell cohort. PMID- 15356124 TI - Expansion and function of CD8+ T cells expressing Ly49 inhibitory receptors specific for MHC class I molecules. AB - MHC class I-specific Ly49 inhibitory receptors regulate NK cell activation, thereby preventing autologous damage to normal cells. Ly49 receptors are also expressed on a subset of CD8+ T cells whose origin and function remain unknown. We report here that, despite their phenotypic and cytolytic similarities, Ly49+CD8+ T cells and conventional Ly49-CD44high memory-phenotype CD8+ T cells present strikingly distinct features. First, under steady state conditions Ly49+CD8+ T cells are poor cytokine producers (TNF-alpha and IFN-gamma) upon TCR triggering. Second, Ly49+CD8+ T cells are not induced upon various settings of Ag immunization or microbial challenge. However, Ly49 can be induced on a fraction of self-specific CD8+ T cells if CD4+ T cells are present. Finally, the size of the Ly49+CD8+ T cell subset is selectively reduced in the absence of STAT1. These results indicate that Ly49 expression is associated with a differentiation program of cytolytic CD8+ T cells triggered upon chronic antigenic exposure. They further suggest that the size of the Ly49+CD8+ T cell subset marks a history of CD8+ T cell activation that might preferentially result from endogenous inducers of inflammation rather than from microbial infections. PMID- 15356125 TI - Sphingosine contributes to glucocorticoid-induced apoptosis of thymocytes independently of the mitochondrial pathway. AB - During the selection process in the thymus, most thymocytes are eliminated by apoptosis through signaling via TCR or glucocorticoids. The involvement of ceramide (Cer) and sphingosine (SP), important apoptotic mediators, remains poorly defined in glucocorticoid-induced apoptosis. We report that, in mouse thymocytes, apoptosis triggered by 10(-6) M dexamethasone (DX) was preceded by a caspase-dependent Cer and SP generation, together with activation of acidic and neutral ceramidases. Apoptosis was drastically reduced by blocking either sphingolipid production (by acid sphingomyelinase inhibitor) or SP production (by ceramidase inhibitors), but not by inhibition of de novo Cer synthesis. Thus, SP generated through acid sphingomyelinase and ceramidase activity would contribute to the apoptotic effect of DX. Consistent with this hypothesis, SP addition or inhibition of SP kinase induced thymocyte apoptosis. DX induced a proteasome dependent loss of mitochondrial membrane potential (Deltapsim) and caspase-8, -3, and -9 processing. Apoptosis was abolished by inhibition of Deltapsim loss or caspase-8 or -3, but not caspase-9. Deltapsim loss was independent of SP production and caspase-8, -3, and -9 activation. However, inhibition of SP production reduced caspase-8 and -3, but not caspase-9 processing. Proteasome inhibition impaired activation of the three caspases, whereas inhibition of Deltapsim loss solely blocked caspase-9 activation. These data indicate that DX induced apoptosis is mediated in part by SP, which contributes, together with proteasome activity, to caspase-8-3 processing independently of mitochondria, and in part by the proteasome/mitochondria pathway, although independently of caspase 9 activation. PMID- 15356126 TI - Enhanced pathogenicity of diabetogenic T cells escaping a non-MHC gene-controlled near death experience. AB - For unknown reasons, the common MHC class I variants encoded by the H2g7 haplotype (Kd, Db) aberrantly elicit autoreactive CD8 T cell responses essential to type 1 diabetes development when expressed in NOD mice, but not other strains. In this study, we show that interactive non-MHC genes allow a NOD-derived diabetogenic CD8 T cell clonotype (AI4) to be negatively selected at far greater efficiency in C57BL/6 mice congenically expressing H2g7 (B6.H2g7). However, the few AI4 T cells escaping negative selection in B6.H2g7 mice are exported from the thymus more efficiently, and are more functionally aggressive than those of NOD origin. This provides mechanistic insight to previous findings that resistant mouse strains carry some genes conferring greater diabetes susceptibility than the corresponding NOD allele. In the B6.H2g7 stock, non-MHC gene-controlled elevations in TCR expression are associated with both enhanced negative selection of diabetogenic CD8 T cells and increased aggressiveness of those escaping this process. An implication of this finding is that the same phenotype, in this case relatively high TCR expression levels, could have double-edged sword effects, contributing to type 1 diabetes resistance at one level of T cell development, but at another actually promoting pathogenesis. PMID- 15356127 TI - A role for the granzyme B inhibitor serine protease inhibitor 6 in CD8+ memory cell homeostasis. AB - Generation and maintenance of protective immunological memory is the goal of vaccination programs. It has recently become clear that CD8+ memory T cells are derived directly from CTLs. The mechanisms underlying this transformation and the subsequent survival of memory cells are not completely understood. However, some effector molecules required by CTLs to eliminate infected cells have also been shown to control the number of Ag-specific cells. We report that memory cells express high levels of serine protease inhibitor (Spi) 6, an inhibitor of the effector molecule granzyme B, and that Spi6 can protect T cells from granzyme B mediated apoptosis. In mouse models, both elevated expression of Spi6 and the complete absence of granzyme B in CD8+ T cells led to an increase in memory cells after infection with lymphocytic choriomeningitis virus. This was not the result of increased levels of antilymphocytic choriomeningitis virus CD8+ T cells during the expansion or contraction phases, but rather transgenic Spi6 directly influenced the survival of CD8+ memory T cells. We propose that expression of protective molecules, like Spi6, serves to shield metabolically active CD8+ memory T cells from their own effector molecules. PMID- 15356128 TI - Enhanced responses of glycosylphosphatidylinositol anchor-deficient T lymphocytes. AB - The functions of GPI-anchored proteins in T lymphocyte activation have been controversial. This issue was addressed by studying the responses of T lymphocytes from T lymphocyte-specific GPI anchor-deficient mice to different stimuli that normally allow coligation of TCR and GPI-anchored proteins. Stimulation of GPI anchor-deficient T lymphocytes with ConA induced 2-fold higher proliferative responses than did normal cells. In response to allogeneic stimulation, proliferation of GPI anchor-deficient T lymphocytes was enhanced 2- to 3-fold. The response to ConA of a GPI anchor-deficient anti-OVA T lymphocyte clone generated from these mice was approximately 3-fold higher than that of cells from the same clone in which GPI anchor expression was restored by retroviral transduction. The response of the GPI anchor-deficient cloned anti-OVA T lymphocytes to antigenic stimulation was similar to that of the retrovirally restored cells. These results indicate that coligation with GPI-anchored proteins counteracts the response to TCR stimulation by ConA or alloantigen but not protein Ag. PMID- 15356129 TI - Expression of the glucocorticoid receptor from the 1A promoter correlates with T lymphocyte sensitivity to glucocorticoid-induced cell death. AB - Glucocorticoid (GC) hormones cause pronounced T cell apoptosis, particularly in immature thymic T cells. This is possibly due to tissue-specific regulation of the glucocorticoid receptor (GR) gene. In mice the GR gene is transcribed from five separate promoters designated: 1A, 1B, 1C, 1D, and 1E. Nearly all cells express GR from promoters 1B-1E, but the activity of the 1A promoter has only been reported in the whole thymus or lymphocyte cell lines. To directly assess the role of GR promoter use in sensitivity to glucocorticoid-induced cell death, we have compared the activity of the GR 1A promoter with GC sensitivity in different mouse lymphocyte populations. We report that GR 1A promoter activity is restricted to thymocyte and peripheral lymphocyte populations and the cortex of the brain. The relative level of expression of the 1A promoter to the 1B-1E promoters within a lymphocyte population was found to directly correlate with susceptibility to GC-induced cell death, with the extremely GC-sensitive CD4+CD8+ thymocytes having the highest levels of GR 1A promoter activity, and the relatively GC-resistant alphabetaTCR+CD24(int/low) thymocytes and peripheral T cells having the lowest levels. DNA sequencing of the mouse GR 1A promoter revealed a putative glucocorticoid-response element. Furthermore, GR 1A promoter use and GR protein levels were increased by GC treatment in thymocytes, but not in splenocytes. These data suggest that tissue-specific differences in GR promoter use determine T cell sensitivity to glucocorticoid-induced cell death. PMID- 15356130 TI - Human galectin-2: novel inducer of T cell apoptosis with distinct profile of caspase activation. AB - Galectin-2 is structurally closely related to galectin-1, but has a distinct expression profile primarily confined to the gastrointestinal tract. Prominent differences in the proximal promoter regions between galectins-2 and -1 concern Sp1-, hepatocyte NF-3, and T cell-specific factor-1 binding sites. Of note, these sequence elements are positioned equally in the respective regions for human and rat galectins-2. Labeled galectin-2 binds to T cells in a beta-galactoside specific manner. In contrast to galectin-1, the glycoproteins CD3 and CD7 are not ligands, while the shared affinity to beta1 integrin (or a closely associated glycoprotein) accounts for a substantial extent of cell surface binding. The carbohydrate-dependent binding of galectin-2 induces apoptosis in activated T cells. Fluorogenic substrate and inhibitor assays reveal involvement of caspases 3 and -9, in accordance with cleavage of the DNA fragmentation factor. Enhanced cytochrome c release, disruption of the mitochondrial membrane potential, and an increase of the Bax/Bcl-2 ratio by opposite regulation of expression of both proteins add to the evidence that the intrinsic apoptotic pathway is triggered. Cell cycle distribution and expression of regulatory proteins remained unaffected. Notably, galectins-1 and -7 reduce cyclin B1 expression, defining functional differences between the structurally closely related galectins. Cytokine secretion of activated T cells was significantly shifted to the Th2 profile. Our study thus classifies galectin-2 as proapoptotic effector for activated T cells, raising a therapeutic perspective. Of importance for understanding the complex galectin network, it teaches the lesson that selection of cell surface ligands, route of signaling, and effects on regulators of cell cycle progression are markedly different between structurally closely related galectins. PMID- 15356131 TI - HIV-1 tat protein modulates the generation of cytotoxic T cell epitopes by modifying proteasome composition and enzymatic activity. AB - Tat, the trans activation protein of HIV, is produced early upon infection to promote and expand HIV replication and transmission. However, Tat appears to also have effects on target cells, which may affect Ag recognition both during infection and after vaccination. In particular, Tat targets dendritic cells and induces their maturation and Ag-presenting functions, increasing Th1 T cell responses. We show in this work that Tat modifies the catalytic subunit composition of immunoproteasomes in B and T cells either expressing Tat or treated with exogenous biological active Tat protein. In particular, Tat up regulates latent membrane protein 7 and multicatalytic endopeptidase complex like 1 subunits and down-modulates the latent membrane protein 2 subunit. These changes correlate with the increase of all three major proteolytic activities of the proteasome and result in a more efficient generation and presentation of subdominant MHC-I-binding CTL epitopes of heterologous Ags. Thus, Tat modifies the Ag processing and modulates the generation of CTL epitopes. This may have an impact on both the control of virally infected cells during HIV-1 infection and the use of Tat for vaccination strategies. PMID- 15356132 TI - IL-6 regulates in vivo dendritic cell differentiation through STAT3 activation. AB - Dendritic cells (DCs) orchestrate immune responses according to their state of maturation. In response to infection, DCs differentiate into mature cells that initiate immune responses, while in the absence of infection, most of them remain in an immature form that induces tolerance to self Ags. Understanding what controls these opposing effects is an important goal for vaccine development and prevention of unwanted immune responses. A crucial question is what cytokine(s) regulates DC maturation in the absence of infection. In this study, we show that IL-6 plays a major role in maintaining immature DCs. IL-6 knockout (KO) mice had increased numbers of mature DCs, indicating that IL-6 blocks DC maturation in vivo. We examined this effect further in knockin mice expressing mutant versions of the IL-6 signal transducer gp130, with defective signaling through either Src homology region 2 domain-containing phosphatase 2/Gab/MAPK (gp130(F759/F759)) or STAT3 (gp130(FxxQ/FxxQ)), and combined gp130 and IL-6 defects (gp130(F759/F759)/IL-6 KO mice). Importantly, we found STAT3 activation by IL-6 was required for the suppression of LPS-induced DC maturation. In addition, STAT3 phosphorylation in DCs was regulated by IL-6 in vivo, and STAT3 was necessary for the IL-6 suppression of bone marrow-derived DC activation/maturation. DC-mediated T cell activation was enhanced in IL-6 KO mice and suppressed in gp130(F759/F759) mice. IL-6 is thus a potent regulator of DC differentiation in vivo, and IL-6 gp130-STAT3 signaling in DCs may represent a critical target for controlling T cell-mediated immune responses in vivo. PMID- 15356133 TI - CD8 T cell responses to lymphocytic choriomeningitis virus in early growth response gene 1-deficient mice. AB - Previous in vitro work has implicated a role for transcriptional factor early growth response gene 1 (EGR1) in regulating immune responses. However, the in vivo role of EGR1 in orchestrating T cell responses has not been studied. To investigate the importance of EGR1 in T cell immunity, we compared Ag-specific CD8 T cell responses between wild type (+/+) and EGR1-deficient (EGR1-/-) mice following an acute infection with lymphocytic choriomeningitis virus (LCMV). These studies revealed that the expansion of LCMV-specific CD8 T cells was substantially reduced in EGR1-/- mice, as compared with +/+ mice. The reduced numbers of LCMV-specific CD8 T cells in EGR1-/- mice were not due to an intrinsic T cell defect per se because purified EGR1-deficient T cells exhibited normal proliferative response to anti-CD3 stimulation in vitro, and underwent normal activation and expansion in response to LCMV upon adoptive transfer into T cell deficient mice. Furthermore, adoptive transfer of CD8 T cells bearing a transgenic TCR into EGR1-/- mice showed that EGR1 deficiency in non-CD8 T cells impaired CD8 T cell expansion in vivo following an LCMV infection. Further investigations on accessory cells showed that bone marrow-derived dendritic cells from EGR1-/- mice did not exhibit detectable impairment to prime Ag-specific CD8 T cell responses in vivo. However, in LCMV-infected mice, EGR1 deficiency selectively impaired the maturation of CD8alpha(+ve) plasmacytoid dendritic cells. Taken together, our findings suggest that EGR1 might promote expansion of CD8 T cells during an acute viral infection by modulating the cues in the lymphoid microenvironment. PMID- 15356134 TI - Impaired type I IFN-induced Jak/STAT signaling in FA-C cells and abnormal CD4+ Th cell subsets in Fancc-/- mice. AB - The Fanconi anemia (FA) group C protein, FANCC, interacts with STAT1 following stimulation with IFN-gamma and is required for proper docking of STAT1 at the IFN gamma receptor alpha-chain (IFN-gammaRalpha, IFN-gammaR1). Consequently, loss of a functional FANCC results in decreased activation of STAT1 following IFN-gamma stimulation. Because type I IFN receptors influence the function of type II receptors, and vice versa, we conducted experiments designed to determine whether type I IFN-induced activation of other STAT proteins is compromised in FA-C cells and found that activation of STAT 1, 3, and 5 is diminished in type I IFN stimulated cells bearing Fancc-inactivating mutations. We also determined that the reduced activation of STATs was accompanied by significant reduction of type I IFN-induced tyrosine kinase 2 and Jak1 phosphorylation. Because tyrosine kinase 2 plays a role in differentiation of Th cells, we quantified cytokine secretion from CD4+ cells and in vitro generated CD4+ Th cell subsets from splenocytes of Fancc null mice to that of heterozygous mice and discovered reduced CD4+ IFN gamma secretion in the Fancc-/- mouse, indicating impaired Th1 differentiation. We suggest that Fancc mutations result in a subtle immunological defect owing to the failure of FANCC to normally support Jak/STAT signaling. PMID- 15356135 TI - An indispensable role for STAT1 in IL-27-induced T-bet expression but not proliferation of naive CD4+ T cells. AB - IL-27 is a novel IL-12 family member that plays a role in the early regulation of Th1 initiation, induces proliferation of naive CD4+ T cells, and synergizes with IL-12 in IFN-gamma production. It has been recently reported that IL-27 induces T bet and IL-12Rbeta2 expression through JAK1/STAT1 activation. In the present study, we further investigated the JAK/STAT signaling molecules activated by IL 27 and also the role of STAT1 in IL-27-mediated responses using STAT1-deficient mice. In addition to JAK1 and STAT1, IL-27-activated JAK2, tyrosine kinase-2, and STAT2, -3, and -5 in naive CD4+ T cells. The activation of STAT2 and STAT5, but not of STAT3, was greatly diminished in STAT1-deficient naive CD4+ T cells. Comparable proliferative response to IL-27 was observed between STAT1-deficient and wild-type naive CD4+ T cells. In contrast, IL-27 hardly induced T-bet and subsequent IL-12Rbeta2 expression, and synergistic IFN-gamma production by IL-27 and IL-12 was impaired in STAT1-deficient naive CD4+ T cells. Moreover, IL-27 augmented the expression of MHC class I on naive CD4+ T cells in a STAT1 dependent manner. These results suggest that IL-27 activates JAK1 and -2, tyrosine kinase-2, STAT1, -2, -3, and -5 in naive CD4+ T cells and that STAT1 plays an indispensable role in IL-27-induced T-bet and subsequent IL-12Rbeta2 expression and MHC class I expression as well but not proliferation, while STAT3 presumably plays an important role in IL-27-induced proliferation. PMID- 15356136 TI - Unraveling the consecutive recombination events in the human IGK locus. AB - In addition to the classical Vkappa-Jkappa, Vkappa-kappa deleting element (Kde), and intron-Kde gene rearrangements, atypical recombinations involving Jkappa recombination signal sequence (RSS) or intronRSS elements can occur in the Igkappa (IGK) locus, as observed in human B cell malignancies. In-depth analysis revealed that atypical JkappaRSS-intronRSS, Vkappa-intronRSS, and JkappaRSS-Kde recombinations not only occur in B cell malignancies, but rather reflect physiological gene rearrangements present in normal human B cells as well. Excision circle analysis and recombination substrate assays can discriminate between single-step vs multistep rearrangements. Using this combined approach, we unraveled that the atypical Vkappa-intronRSS and JkappaRSS-Kde pseudohybrid joints most probably result from ongoing recombination following an initial aberrant JkappaRSS-intronRSS signal joint formation. Based on our observations in normal and malignant human B cells, a model is presented to describe the sequential (classical and atypical) recombination events in the human IGK locus and their estimated relative frequencies (0.2-1.0 vs < 0.03). The initial JkappaRSS-intronRSS signal joint formation (except for Jkappa1RSS-intronRSS) might be a side event of an active V(D)J recombination mechanism, but the subsequent formation of Vkappa-intronRSS and JkappaRSS-Kde pseudohybrid joints can represent an alternative pathway for IGK allele inactivation and allelic exclusion, in addition to classical Ckappa deletions. Although usage of this alternative pathway is limited, it seems essential for inactivation of those IGK alleles that have undergone initial aberrant recombinations, which might otherwise hamper selection of functional Ig L chain proteins. PMID- 15356137 TI - Dual function of the extracellular matrix: stimulatory for cell cycle progression of naive T cells and antiapoptotic for tissue-derived memory T cells. AB - Tissue T cells encounter Ag in a distinct microenvironment, where they are embedded in the interstitial extracellular matrix (ECM). In contrast, while naive T cells are exposed to Ag in the lymph node, immediately after naive T cells are activated they must extravasate into the ECM to function effectively. Because integrin-mediated adhesion to the ECM modulates cell cycle progression and survival in adherent nonimmune cells, we hypothesize that blood and tissue derived T cells have similarly adapted their behavior to their first or continued encounter with ECM. T cells from peripheral blood (PBT) and tissue (the intestinal lamina propria T cell (LPT)) were stimulated with anti-CD3-coated beads in the presence or absence of native ECM derived from intestinal fibroblasts, plate-immobilized fibronectin, or collagen type I. Native ECM and collagen, but not fibronectin, induced in anti-CD3 activated PBT a 4- to 5-fold increase in the entry, progression, and completion of the cell cycle over that triggered by anti-CD3 alone. Neutralizing beta1 integrin Abs abrogated this increase. None of these ECM proteins stimulated cell cycle progression in LPT. In contrast, anti-CD3 activation of LPT in the presence of native ECM and fibronectin reduced activation-induced cell death by 40%. These results demonstrate that naive and effector/memory T cells respond differently upon exposure to specific ECM components. When naive PBT encounter Ag in the context of ECM, their progression through the cell cycle is enhanced, favoring clonal expansion; while tissue T cell longevity may be mediated by interactions with the ECM. PMID- 15356138 TI - Signaling through CD70 regulates B cell activation and IgG production. AB - CD70, the cellular ligand of the TNF receptor family member CD27, is expressed transiently on activated T and B cells and constitutively on a subset of B cell chronic lymphocytic leukemia and large B cell lymphomas. In the present study, we used B cells constitutively expressing CD70 to study the functional consequences of signaling through CD70. In vitro, CD70 ligation with anti-CD70 mAbs strongly supported proliferation and cell cycle entry of B cells submitogenically stimulated with either anti-CD40 mAb, LPS, or IL-4. In this process, the cell surface receptors CD25, CD44, CD69, CD95, and GL7 were up-regulated, whereas the expression of CD21, CD62L, surface IgM (sIgM), and sIgD was decreased. Addition of CD70 mAb to low dose LPS-stimulated CD70-positive B cells strongly diminished IgG secretion and enhanced production of IgM. Signaling through CD70 on B cells was dependent on the initiation of both PI3K and MEK pathways. In vivo exposure to either CD70 mAb or the CD70 counterreceptor CD27 down-regulated CD62L and sIgM on CD70-positive B cells. CD70 signaling during T cell-dependent immune responses also decreased IgG-specific Ab titers. Together, the in vitro and in vivo data demonstrate that CD70 has potent reverse signaling properties in B cells, initiating a signaling cascade that regulates expansion and differentiation. PMID- 15356139 TI - A role for CD28 in lymphopenia-induced proliferation of CD4 T cells. AB - The peripheral mechanisms that regulate the size and the repertoire of the T cell compartment during recovery from a lymphopenic state are incompletely understood. In particular, the role of costimulatory signals, such as those provided by CD28, which have a critical importance for the immune response toward foreign Ags in nonlymphopenic animals, has been unclear in lymphopenia-induced proliferation (LIP). In this study, we show that accumulation of highly divided CD4 T cells characterized by great potential to make IFN-gamma is significantly delayed in the absence of B7:CD28 costimulation during LIP. Furthermore, CD28-sufficient CD4 T cells show great competitive advantage over CD28-deficient CD4 T cells when transferred together into the same lymphopenic hosts. Administration of CTLA-4-Ig removed this competitive advantage. Interestingly, CTLA-4-Ig treatment resulted in modest inhibition of LIP by CD28-deficient responders, suggesting that some of its effects may be independent of mere B7 blockade. PMID- 15356140 TI - Microglia initiate central nervous system innate and adaptive immune responses through multiple TLRs. AB - Microglia are the resident macrophage-like population in the CNS. Microglia remain quiescent until injury or infection activates the cells to perform effector inflammatory and APC functions. Our previous studies have shown that microglia infected with a neurotropic strain of Theiler's murine encephalomyelitis virus secreted innate immune cytokines and up-regulated costimulatory molecules and MHC class II, enabling the cells to present viral and myelin Ags to CD4+ T cells. Recently, TLRs have been shown to recognize pathogen associated molecular patterns and initiate innate immune responses upon interaction with infectious agents. We examined TLR expression on brain microglia and their functional responses upon stimulation with various TLR agonists. We report that mouse microglia express mRNA for all of the recently identified TLRs, TLR1-9, used for recognition of bacterial and viral molecular patterns. Furthermore, stimulation of quiescent microglia with various TLR agonists, including LPS (TLR4), peptidoglycan (TLR2), polyinosinic-polycytidylic acid (TLR3), CpG DNA (TLR9), and infection with viable Theiler's murine encephalomyelitis virus, activated the cells to up-regulate unique patterns of innate and effector immune cytokines and chemokines at the mRNA and protein levels. In addition, TLR stimulation activated up-regulation of MHC class II and costimulatory molecules, enabling the microglia to efficiently present myelin Ags to CD4+ T cells. Thus, microglia appear to be a unique and important component of both the innate and adaptive immune response, providing the CNS with a means to rapidly and efficiently respond to a wide variety of pathogens. PMID- 15356141 TI - Immunoproteasomes down-regulate presentation of a subdominant T cell epitope from lymphocytic choriomeningitis virus. AB - The cytotoxic T cell response to pathogens is usually directed against a few immunodominant epitopes, while other potential epitopes are either subdominant or not used at all. In C57BL/6 mice, the acute cytotoxic T cell response against lymphocytic choriomeningitis virus is directed against immunodominant epitopes derived from the glycoprotein (gp33-41) and the nucleoprotein (NP396-404), while the gp276-286 epitope remains subdominant. Despite extensive investigations, the reason for this hierarchy between epitopes is not clear. In this study, we show that the treatment of cells with IFN-gamma enhanced the presentation of gp33-41, whereas presentation of the gp276-286 epitope from the same glycoprotein was markedly reduced. Because proteasomes are crucially involved in epitope generation and because IFN-gamma treatment in vitro and lymphocytic choriomeningitis virus infection in vivo lead to a gradual replacement of constitutive proteasomes by immunoproteasomes, we investigated the role of proteasome composition on epitope hierarchy. Overexpression of the active site subunits of immunoproteasomes LMP2, LMP7, and MECL-1 as well as overexpression of LMP2 alone suppressed the presentation of the gp276-286 epitope. The ability to generate gp276-286-specific CTLs was enhanced in LMP2- and LMP7-deficient mice, and macrophages from these mice showed an elevated presentation of this epitope. In vitro digests demonstrated that fragmentation by immunoproteasomes, but not constitutive proteasomes led to a preferential destruction of the gp276 epitope. Taken together, we show that LMP2 and LMP7 can at least in part determine subdominance and shape the epitope hierarchy of CTL responses in vivo. PMID- 15356142 TI - Analysis of Notch1 function by in vitro T cell differentiation of Pax5 mutant lymphoid progenitors. AB - Signaling through the Notch1 receptor is essential for T cell development in the thymus. Stromal OP9 cells ectopically expressing the Notch ligand Delta-like1 mimic the thymic environment by inducing hemopoietic stem cells to undergo in vitro T cell development. Notch1 is also expressed on Pax5-/- pro-B cells, which are clonable lymphoid progenitors with a latent myeloid potential. In this study, we demonstrate that Pax5-/- progenitors efficiently differentiate in vitro into CD4+CD8+ alphabeta and gammadelta T cells upon coculture with OP9-Delta-like1 cells. In vitro T cell development of Pax5-/- progenitors strictly depends on Notch1 function and progresses through normal developmental stages by expressing T cell markers and rearranging TCRbeta, gamma, and delta loci in the correct temporal sequence. Notch-stimulated Pax5-/- progenitors efficiently down-regulate the expression of B cell-specific genes, consistent with a role of Notch1 in preventing B lymphopoiesis in the thymus. At the same time, Notch signaling rapidly induces cell surface expression of the c-Kit receptor and transcription of the target genes Deltex1 and pre-Talpha concomitant with the activation of TCR Vbeta germline transcription and the regulatory genes GATA3 and Tcf1. These data suggest that Notch1 acts upstream of GATA3 and Tcf1 in early T cell development and regulates Vbeta-DJbeta rearrangements by controlling the chromatin accessibility of Vbeta genes at the TCRbeta locus. PMID- 15356143 TI - Delta-short consensus repeat 4-decay accelerating factor (DAF: CD55) inhibits complement-mediated cytolysis but not NK cell-mediated cytolysis. AB - NK cells play a critical role in the rejection of xenografts. In this study, we report on an investigation of the effect of complement regulatory protein, a decay accelerating factor (DAF: CD55), in particular, on NK cell-mediated cytolysis. Amelioration of human NK cell-mediated pig endothelial cell (PEC) and pig fibroblast cell lyses by various deletion mutants and point substitutions of DAF was tested, and compared with their complement regulatory function. Although wild-type DAF and the delta-short consensus repeat (SCR) 1-DAF showed clear inhibition of both complement-mediated and NK-mediated PEC lyses, delta-SCR2-DAF and delta-SCR3-DAF failed to suppress either process. However, delta-SCR4-DAF showed a clear complement regulatory effect, but had no effect on NK cells. Conversely, the point substitution of DAF (L147 x F148 to SS and KKK(125-127) to TTT) was half down-regulated in complement inhibitory function, but the inhibition of NK-mediated PEC lysis remained unchanged. Other complement regulatory proteins, such as the cell membrane-bound form factor H, fH-PI, and C1 inactivator, C1-INH-PI, and CD59 were also assessed, but no suppressive effect on NK cell-mediated PEC lysis was found. These data suggest, for DAF to function on NK cells, SCR2-4 is required but no relation to its complement regulatory function exists. PMID- 15356144 TI - The murine NK receptor 2B4 (CD244) exhibits inhibitory function independent of signaling lymphocytic activation molecule-associated protein expression. AB - 2B4 (CD244) is a receptor belonging to the CD2-signaling lymphocytic activation molecule family and is found on all murine NK cells and a subset of NKT and CD8+ T cells. Murine 2B4 is expressed as two isoforms (2B4 short and 2B4 long) that arise by alternative splicing. They differ only in their cytoplasmic domains and exhibit opposing function when expressed in the RNK-16 cell line. The ligand for 2B4, CD48, is expressed on all hemopoietic cells. Previous studies have shown that treatment of NK cells with a 2B4 mAb results in increased cytotoxicity and IFN-gamma production. In this report, we used CD48+/- variants of the P815 tumor cell line and 2B4 knockout mice to show that engagement of 2B4 by its counterreceptor, CD48, expressed on target cells leads to an inhibition in NK cytotoxicity. The addition of 2B4 or CD48 mAb relieves this inhibition resulting in enhanced target cell lysis. This 2B4-mediated inhibition acts independently of signaling lymphocytic activation molecule-associated protein expression. Imaging studies show that 2B4 preferentially accumulates at the interface between NK and target cells during nonlytic events also indicative of an inhibitory receptor. This predominant inhibitory function of murine 2B4 correlates with increased 2B4 long isoform level expression over 2B4 short. PMID- 15356145 TI - Phosphorylation of Grb2-associated binder 2 on serine 623 by ERK MAPK regulates its association with the phosphatase SHP-2 and decreases STAT5 activation. AB - IL-2 stimulation of T lymphocytes induces the tyrosine phosphorylation and adaptor function of the insulin receptor substrate/Grb2-associated binder (Gab) family member, Gab2. In addition, Gab2 undergoes a marked decrease in its mobility in SDS-PAGE, characteristic of migration shifts induced by serine/threonine phosphorylations in many proteins. This migration shift was strongly diminished by treating cells with the MEK inhibitor U0126, indicating a possible role for ERK in Gab2 phosphorylation. Indeed, ERK phosphorylated Gab2 on a consensus phosphorylation site at serine 623, a residue located between tyrosine 614 and tyrosine 643 that are responsible for Gab2/Src homology 2 domain containing tyrosine phosphatase (SHP)-2 interaction. We report that pretreatment of Kit 225 cells with U0126 increased Gab2/SHP-2 association and tyrosine phosphorylation of SHP-2 in response to IL-2, suggesting that ERK phosphorylation of serine 623 regulates the interaction between Gab2 and SHP-2, and consequently the activity of SHP-2. This hypothesis was confirmed by biochemical analysis of cells expressing Gab2 WT, Gab2 serine 623A or Gab2 tyrosine 614F, a mutant that cannot interact with SHP-2 in response to IL-2. Activation of the ERK pathway was indeed blocked by Gab2 tyrosine 614F and slightly increased by Gab2 serine 623A. In contrast, STAT5 activation was strongly enhanced by Gab2 tyrosine 614F, slightly reduced by Gab2 WT and strongly inhibited by Gab2 serine 623A. Analysis of the rate of proliferation of cells expressing these mutants of Gab2 demonstrated that tyrosine 614F mutation enhanced proliferation whereas serine 623A diminished it. These results demonstrate that ERK-mediated phosphorylation of Gab2 serine 623 is involved in fine tuning the proliferative response of T lymphocytes to IL-2. PMID- 15356146 TI - The binding sites for competitive antagonistic, allosteric antagonistic, and agonistic antibodies to the I domain of integrin LFA-1. AB - We explore the binding sites for mAbs to the alpha I domain of the integrin alphaLbeta2 that can competitively inhibit, allosterically inhibit, or activate binding to the ligand ICAM-1. Ten mAbs, some of them clinically important, were mapped to species-specific residues. The results are interpreted with independent structures of the alphaL I domain determined in seven different crystal lattices and in solution, and which are present in three conformational states that differ in affinity for ligand. Six mAbs bind to adjacent regions of the beta1-alpha1 and alpha3-alpha4 loops, which show only small (mean, 0.8 angstroms; maximum, 1.8 angstroms) displacements among the eight I domain structures. Proximity to the ligand binding site and to noncontacting portions of the ICAM-1 molecule explains competitive inhibition by these mAbs. Three mAbs bind to a segment of seven residues in the beta5-alpha6 loop and alpha6 helix, in similar proximity to the ligand binding site, but on the side opposite from the beta1-alpha1/alpha3-alpha4 epitopes, and far from noncontacting portions of ICAM-1. These residues show large displacements among the eight structures in response to lattice contacts (mean, 3.6 angstroms; maximum, 9.4 angstroms), and movement of a buried Phe in the beta5-alpha6 loop is partially correlated with affinity change at the ligand binding site. Together with a lack of proximity to noncontacting portions of ICAM 1, these observations explain variation among this group of mAbs, which can either act as competitive or allosteric antagonists. One agonistic mAb binds distant from the ligand binding site of the I domain, to residues that show little movement (mean, 0.5 angstroms; maximum, 1.0 angstroms). Agonism by this mAb is thus likely to result from altering the orientation of the I domain with respect to other domains within an intact integrin alphaLbeta2 heterodimer. PMID- 15356147 TI - Histone deacetylase 3, a class I histone deacetylase, suppresses MAPK11-mediated activating transcription factor-2 activation and represses TNF gene expression. AB - During inflammatory events, the induction of immediate-early genes, such as TNF alpha, is regulated by signaling cascades including the JAK/STAT, NF-kappaB, and the p38 MAPK pathways, which result in phosphorylation-dependent activation of transcription factors. We observed the direct interaction of histone deacetylase (HDAC) 3, a class I histone deacetylase, with MAPK11 (p38 beta isoform) by West Western-based screening analysis, pull-down assay, and two-hybrid system analysis. Results further indicated that HDAC3 decreases the MAPK11 phosphorylation state and inhibits the activity of the MAPK11-dependent transcription factor, activating transcription factor-2 (ATF-2). LPS-mediated activation of ATF-2 was inhibited by HDAC3 in a time- and dose-dependent manner. Inhibition of HDAC3 expression by RNA interference resulted in increased ATF-2 activation in response to LPS stimulation. In agreement with decreased ATF-2 transcriptional activity by HDAC3, HDAC3-repressed TNF gene expression, and TNF protein production observed in response to LPS stimulation. Therefore, our results indicate that HDAC3 interacts directly and selectively with MAPK11, represses ATF-2 transcriptional activity, and acts as a regulator of TNF gene expression in LPS-stimulated cells, especially in mononuclear phagocytes. PMID- 15356148 TI - Characterization of a dipeptide motif regulating IFN-gamma receptor 2 plasma membrane accumulation and IFN-gamma responsiveness. AB - The IFN-gammaR complex is composed of two IFN-gammaR1 and two IFN-gammaR2 polypeptide chains. Although IFN-gammaR1 is constitutively expressed on all nucleated cells, IFN-gammaR2 membrane display is selective and tightly regulated. We created a series of fluorescent-tagged IFN-gammaR2 expression constructs to follow the molecule's cell surface expression and intracellular distribution. Truncation of the receptor immediately upstream of Leu-Ile 255-256 (254X) created a receptor devoid of signaling that overaccumulated on the cell surface. In addition, this truncated receptor inhibited wild-type IFN-gammaR2 activity and therefore exerted a dominant negative effect. In-frame deletion (255Delta2) or alanine substitution (LI255-256AA) of these amino acids created mutants that overaccumulated on the plasma membrane, but had enhanced function. Single amino acid substitutions (L255A or I256A) had a more modest effect. In-frame deletions upstream (253Delta2), but not downstream (257Delta2), of Leu-Ile 255-256 also led to overaccumulation. A truncation within the IFN-gammaR2 Jak2 binding site (270X) led to a mutant devoid of function that did not overaccumulate and did not affect wild-type IFN-gammaR2 signaling. We have created a series of novel mutants of IFN gammaR2 that have facilitated the identification of intracellular domains that control IFN-gammaR2 accumulation and IFN-gamma responsiveness. In contrast to IFN gammaR1, not only dominant negative, but also dominant gain-of-function, mutations were created through manipulation of IFN-gammaR2 Leu-Ile 255-256. These IFN-gammaR2 mutants will allow fine dissection of the role of IFN-gamma signaling in immunity. PMID- 15356149 TI - A novel mutation in IFN-gamma receptor 2 with dominant negative activity: biological consequences of homozygous and heterozygous states. AB - We identified two siblings homozygous for a single base pair deletion in the IFN gammaR2 transmembrane domain (791delG) who presented with multifocal Mycobacterium abscessus osteomyelitis (patient 1) and disseminated CMV and Mycobacterium avium complex infection (patient 2), respectively. Although the patients showed no IFN-gammaR activity, their healthy heterozygous parents showed only partial IFN-gammaR activity. An HLA-identical bone marrow transplant from the mother led patient 1 to complete hemopoietic reconstitution, but only partial IFN-gammaR function. We cloned and expressed fluorescent fusion proteins of the wild-type IFN-gammaR2, an IFN-gammaR2 mutant previously described to produce a complete autosomal recessive deficiency (278del2), and of 791delG to determine whether the intermediate phenotype in the 791delG heterozygous state was caused by haploinsufficiency or a dominant negative effect. When cotransfected together with the wild-type vector into IFN-gammaR2-deficient fibroblasts, the fusion protein with 791delG inhibited IFN-gammaR function by 48.7 +/- 5%, whereas fusion proteins with 278del2 had no inhibitory effect. Confocal microscopy of 791delG fusion proteins showed aberrant diffuse intracellular accumulation without plasma membrane localization. The fusion protein created by 791delG did not complete Golgi processing, and was neither expressed on the plasma membrane, nor shed extracellularly. The mutant construct 791delG exerts dominant negative effects on IFN-gamma signaling without cell surface display, suggesting that it is acting on pathways other than those involved in cell surface recognition of ligand. PMID- 15356150 TI - Distinct and opposite activities of human terminal deoxynucleotidyltransferase splice variants. AB - Evidence for potential human TdT (hTdT) isoforms derived from hTdT genomic sequences led us to identify the short isoform (hTdTS), as well as mature long transcripts containing exon XII (hTdTL1) and another including exon VII (hTdTL2) in lymphoid cells. Normal B and T lymphocytes express exclusively hTdTS and hTdTL2, whereas hTdTL1 expression appears to be restricted to transformed lymphoid cell lines. In in vitro recombination and primer assays, both long isoforms were shown to have 3'-->5' exonuclease activity. Overexpression of hTdTS or hTdTL2 greatly reduced the efficiency of recombination, which was reverted to normal levels by the simultaneous expression of both enzymes. Therefore, alternative splicing may prevent the adverse effects of unchecked elongation or diminution of coding ends during V(D)J recombination, thus affecting the survival of a B or T cell precursor during receptor gene rearrangements. Finally, the newly discovered hTdT isoforms should be considered in future screening of human leukemias. PMID- 15356151 TI - IL-13 activates a mechanism of tissue fibrosis that is completely TGF-beta independent. AB - Fibrosis is a characteristic feature in the pathogenesis of a wide spectrum of diseases. Recently, it was suggested that IL-13-dependent fibrosis develops through a TGF-beta1 and matrix metalloproteinase-9-dependent (MMP-9) mechanism. However, the significance of this pathway in a natural disorder of fibrosis was not investigated. In this study, we examined the role of TGF-beta in IL-13 dependent liver fibrosis caused by Schistosoma mansoni infection. Infected IL-13 /- mice showed an almost complete abrogation of fibrosis despite continued and undiminished production of TGF-beta1. Although MMP-9 activity was implicated in the IL-13 pathway, MMP-9-/- mice displayed no reduction in fibrosis, even when chronically infected. To directly test the requirement for TGF-beta, studies were also performed with neutralizing anti-TGF-beta Abs, soluble antagonists (soluble TGF-betaR-Fc), and Tg mice (Smad3-/- and TGF-betaRII-Fc Tg) that have disruptions in all or part of the TGF-beta signaling cascade. In all cases, fibrosis developed normally and with kinetics similar to wild-type mice. Production of IL 13 was also unaffected. Finally, several genes, including interstitial collagens, several MMPs, and tissue inhibitors of metalloprotease-1 were up-regulated in TGF beta1-/- mice by IL-13, demonstrating that IL-13 activates the fibrogenic machinery directly. Together, these studies provide unequivocal evidence of a pathway of fibrogenesis that is IL-13 dependent but TGF-beta1 independent, illustrating the importance of targeting IL-13 directly in the treatment of infection-induced fibrosis. PMID- 15356152 TI - Mechanisms of host defense following severe acute respiratory syndrome coronavirus (SARS-CoV) pulmonary infection of mice. AB - We describe a model of severe acute respiratory syndrome-coronavirus (SARS-CoV) infection in C57BL/6 mice. A clinical isolate of the virus introduced intranasally replicated transiently to high levels in the lungs of these mice, with a peak on day 3 and clearance by day 9 postinfection. Viral RNA localized to bronchial and bronchiolar epithelium. Expression of mRNA for angiotensin converting enzyme 2, the SARS-CoV receptor, was detected in the lung following infection. The virus induced production in the lung of the proinflammatory chemokines CCL2, CCL3, CCL5, CXCL9, and CXCL10 with differential kinetics. The receptors for these chemokines were also detected. Most impressively, mRNA for CXCR3, the receptor for CXCL9 and CXCL10, was massively up-regulated in the lungs of SARS-CoV-infected mice. Surprisingly Th1 (and Th2) cytokines were not detectable, and there was little local accumulation of leukocytes and no obvious clinical signs of pulmonary dysfunction. Moreover, beige, CD1-/-, and RAG1-/- mice cleared the virus normally. Infection spread to the brain as it was cleared from the lung, again without leukocyte accumulation. Infected mice had a relative failure to thrive, gaining weight significantly more slowly than uninfected mice. These data indicate that C57BL/6 mice support transient nonfatal systemic infection with SARS-CoV in the lung, which is able to disseminate to brain. In this species, proinflammatory chemokines may coordinate a rapid and highly effective innate antiviral response in the lung, but NK cells and adaptive cellular immunity are not required for viral clearance. PMID- 15356153 TI - IL-12p40 and IL-18 modulate inflammatory and immune responses to respiratory syncytial virus infection. AB - Respiratory syncytial virus-induced bronchiolitis has been linked to the development of allergy and atopic asthma. IL-12 and possibly IL-18 are central mediators orchestrating Th1 and/or Th2 immune responses to infection. To determine a possible role for IL-12 in regulating the immune response to acute respiratory syncytial virus infection, IL-12p40 gene-targeted (IL-12p40-/-) and wild-type mice were intratracheally infected with respiratory syncytial virus, and lung inflammatory and immune responses were assessed. Lung inflammation and mucus production were increased in the airways of IL-12p40-/- mice as compared with those of wild-type mice, concurrent with increased levels of the Th2 effector cytokines IL-5 and IL-13. Respiratory syncytial virus clearance and levels of Th1 effector cytokine IFN-gamma were not altered. Interestingly, IL-18, another mediator of IFN-gamma production, was significantly increased in the lungs of IL-12p40-/- mice early during the course of infection. Abrogation of IL 18-mediated signaling in IL-12p40-/- mice further enhanced Th2 immune response and mucus production in the airways during respiratory syncytial virus infection but failed to modulate IFN-gamma production or viral clearance. These findings implicate a role for IL-12 and IL-18 in modulating respiratory syncytial virus induced airway inflammation distinct from that of viral clearance. PMID- 15356154 TI - Identification of immunodominant sites on the spike protein of severe acute respiratory syndrome (SARS) coronavirus: implication for developing SARS diagnostics and vaccines. AB - The spike (S) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) is not only responsible for receptor binding and virus fusion, but also a major Ag among the SARS-CoV proteins that induces protective Ab responses. In this study, we showed that the S protein of SARS-CoV is highly immunogenic during infection and immunizations, and contains five linear immunodominant sites (sites I to V) as determined by Pepscan analysis with a set of synthetic peptides overlapping the entire S protein sequence against the convalescent sera from SARS patients and antisera from small animals immunized with inactivated SARS-CoV. Site IV located in the middle region of the S protein (residues 528-635) is a major immunodominant epitope. The synthetic peptide S(603-634), which overlaps the site IV sequence reacted with all the convalescent sera from 42 SARS patient, but none of the 30 serum samples from healthy blood donors, suggesting its potential application as an Ag for developing SARS diagnostics. This study also provides information useful for designing SARS vaccines and understanding the SARS pathogenesis. PMID- 15356155 TI - Salmonella escape from antigen presentation can be overcome by targeting bacteria to Fc gamma receptors on dendritic cells. AB - Dendritic cells (DCs) are professional APCs with the unique ability to activate naive T cells, which is required for initiation of the adaptive immune response against pathogens. Therefore, interfering with DC function would be advantageous for pathogen survival and dissemination. In this study we provide evidence suggesting that Salmonella enterica serovar typhimurium, the causative agent of typhoid disease in the mouse, interferes with DC function. Our results indicate that by avoiding lysosomal degradation, S. typhimurium impairs the ability of DCs to present bacterial Ags on MHC class I and II molecules to T cells. This process could correspond to a novel mechanism developed by this pathogen to evade adaptive immunity. In contrast, when S. typhimurium is targeted to FcgammaRs on DCs by coating bacteria with Salmonella-specific IgG, bacterial Ags are efficiently processed and presented on MHC class I and class II molecules. This enhanced Ag presentation leads to a robust activation of bacteria-specific T cells. Laser confocal microscopy experiments show that virulent S. typhimurium is rerouted to the lysosomal degradation pathway of DCs when internalized through FcgammaR. These observations are supported by electron microscopy studies demonstrating that internalized S. typhimurium shows degradation signs only when coated with IgG and captured by FcgammaRs on DCs. Therefore, our data support a potential role for bacteria-specific IgG on the augmentation of Ag processing and presentation by DCs to T cells during the immune response against intracellular bacteria. PMID- 15356156 TI - Hemozoin (malarial pigment) inhibits differentiation and maturation of human monocyte-derived dendritic cells: a peroxisome proliferator-activated receptor gamma-mediated effect. AB - Acute and chronic Plasmodium falciparum malaria are accompanied by severe immunodepression possibly related to subversion of dendritic cells (DC) functionality. Phagocytosed hemozoin (malarial pigment) was shown to inhibit monocyte functions related to immunity. Hemozoin-loaded monocytes, frequently found in circulation and adherent to endothelia in malaria, may interfere with DC development and play a role in immunodepression. Hemozoin-loaded and unloaded human monocytes were differentiated in vitro to immature DC (iDC) by treatment with GM-CSF and IL-4, and to mature DC (mDC) by LPS challenge. In a second setting, hemozoin was fed to iDC further cultured to give mDC. In both settings, cells ingested large amounts of hemozoin undegraded during DC maturation. Hemozoin-fed monocytes did not apoptose but their differentiation and maturation to DC was severely impaired as shown by blunted expression of MHC class II and costimulatory molecules CD83, CD80, CD54, CD40, CD1a, and lower levels of CD83 specific mRNA in hemozoin-loaded iDC and mDC compared with unfed or latex-loaded DC. Further studies indicated activation of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) in hemozoin-loaded iDC and mDC, associated with increased expression of PPAR-gamma mRNA, without apparent involvement of NF kappaB. Moreover, expression of PPAR-gamma was induced and up-regulation of CD83 was inhibited by supplementing iDC and mDC with plausible concentrations of 15(S) hydroxyeicosatetraenoic acid, a PPAR-gamma ligand abundantly produced by hemozoin via heme-catalyzed lipoperoxidation. PMID- 15356158 TI - The development of functional CD8 T cell memory after Listeria monocytogenes infection is not dependent on CD40. AB - The immunologic requirements for generating long-lived protective CD8 T cell memory remain unclear. Memory CD8 populations generated in the absence of CD4 Th cells reportedly have functional defects, and at least a subset of CD8 T cells transiently express CD40 after activation, suggesting that direct CD4-CD8 T cell interactions through CD40 may influence the magnitude and functional quality of memory CD8 populations. To ascertain the role of CD40 in such direct T cell interactions, we investigated CD8 T cell responses in CD40-/- mice after infection with Listeria monocytogenes, an intracellular bacterium that induces APC activation and thus priming of CD8 T cells independently of CD4 Th cell help through CD40. In this study we show that memory CD8 T cells generated in CD40 deficient mice show in vivo cytotoxicity and cytokine production equivalent to CD8 memory T cells from wild-type mice. Upon secondary Listeria infection, CD40-/ memory CD8 T cells expand to greater numbers than seen in wild-type mice. These results indicate that CD40 ligation on CD8 T cells, although reportedly a part of CD8 T cell memory development in an H-Y-directed response, is not needed for the development of functional memory CD8 T cell populations after Listeria infection. PMID- 15356157 TI - STAT4 is a critical mediator of early innate immune responses against pulmonary Klebsiella infection. AB - Bacterial pneumonia is a leading cause of morbidity and mortality in the U.S. An effective innate immune response is critical for the clearance of bacteria from the lungs. IL-12, a key T1 cytokine in innate immunity, signals through STAT4. Thus, understanding how STAT4 mediates pulmonary immune responses against bacterial pathogens will have important implications for the development of rational immunotherapy targeted at augmenting innate immunity. We intratracheally administered Klebsiella pneumoniae to wild-type BALB/c and STAT4 knockout (STAT4 /-) mice. Compared with wild-type controls, STAT4-/- mice had decreased survival following intratracheal Klebsiella administration, which was associated with a higher lung and blood bacterial burden. STAT4-/- animals also displayed impaired pulmonary IFN-gamma production and decreased levels of proinflammatory cytokines, including the ELR- CXC chemokines IFN-gamma-inducible protein-10 and monokine induced by IFN-gamma. Although total lung leukocyte populations were similar between STAT4-/- and wild-type animals following infection, alveolar macrophages isolated from infected STAT4-/- mice had decreased production of proinflammatory cytokines, including IFN-gamma, compared with infected wild-type mice. The intrapulmonary overexpression of IFN-gamma concomitant with the systemic administration of IFN-gamma partially reversed the immune deficits observed in STAT4-/- mice, resulting in improved bacterial clearance from the blood. Collectively, these studies demonstrate that STAT4 is required for the generation of an effective innate host defense against bacterial pathogens of the lung. PMID- 15356159 TI - Low-dose Salmonella infection evades activation of flagellin-specific CD4 T cells. AB - Many pathogens can establish a lethal infection from relatively small inocula, yet the effect of infectious dose upon CD4 T cell activation is not clearly understood. This issue was examined by tracking Salmonella flagellin-specific SM1 T cells in vivo, after i.v. and oral challenge of mice with virulent Salmonella typhimurium. SM1 T cells rapidly expressed activation markers and expanded in response to high-dose infection but remained completely unresponsive in mice challenged with low doses of Salmonella. SM1 T cells, in these mice, remained unresponsive, despite massive bacterial replication in vivo. Naive SM1 T cells in low-dose Salmonella-infected mice were activated rapidly after the injection of flagellin peptide, demonstrating that these T cells were fully capable of responding, ruling out the possibility of a bacterial-induced suppressive environment. The inability of flagellin-specific SM1 T cells to respond to low dose infection was not due to Ag down-regulation, because flagellin expression was detected using a functional assay. Together, these data suggest that low-dose Salmonella infection can evade flagellin-specific CD4 T cell activation in vivo. PMID- 15356160 TI - Protection against late-onset AIDS in macaques prophylactically immunized with a live simian HIV vaccine was dependent on persistence of the vaccine virus. AB - This is a 5-year follow-up study on 12 macaques that were immunized orally with two live SHIV vaccines, six with V1 and six with V2. All 12 macaques became persistently infected after transient replication of the vaccine viruses; all were challenged vaginally 6 mo later with homologous pathogenic SHIV(KU-1). Two of the V1 group developed full-blown AIDS without evidence of vaccine virus DNA in tissues. The data on the 10 vaccinated survivors showed that all 10 became infected with SHIV(KU-1) and that DNA of both vaccine and SHIV(KU-1) viruses were present 6 mo postchallenge, with minimal replication of SHIV(KU-1). During the following 5 years, these animals remained persistently infected, but with only one of the two viruses. Six animals eliminated their vaccine virus after variable periods of time and four of these succumbed to reactivation of the challenge virus and AIDS. Five years after challenge, four latently infected animals, two with V2 and two with SHIV(KU-1), were reinoculated with SHIV(KU-1.) This resulted in transient superinfection and the animals promptly returned to their prechallenge status. Immunosuppression of the four animals 1 year later with Abs to CD8+ lymphocytes resulted in transiently productive replication of their respective latent viruses, and upon recovery of CD8+ lymphocytes, they reverted to their latent virus status. The major finding was that of eight animals that eliminated the vaccine virus, six developed AIDS. The two others harboring SHIV(KU-1) remain at risk for developing late-onset disease. The primary correlate against AIDS was persistence of the vaccine virus. PMID- 15356161 TI - Herpes simplex virus type-1-induced activation of myeloid dendritic cells: the roles of virus cell interaction and paracrine type I IFN secretion. AB - Adaptive cellular immunity is required to clear HSV-1 infection in the periphery. Myeloid dendritic cells (DCs) are the first professional Ag-presenting cell to encounter the virus after primary and secondary infection and thus the consequences of their infection are important in understanding the pathogenesis of the disease and the response to the virus. Following HSV-1 infection, both uninfected and infected human DCs acquire a more mature phenotype. In this study, we demonstrate that type I IFN secreted from myeloid DC mediates bystander activation of the uninfected DCs. Furthermore, we confirm that this IFN primes DCs for elevated IL-12 p40 and p70 secretion. However, secretion of IFN is not responsible for the acquisition of a mature phenotype by HSV-1-infected DC. Rather, virus binding to a receptor on the cell surface induces DC maturation directly, through activation of the NF-kappaB and p38 MAPK pathways. The binding of HSV glycoprotein D is critical to the acquisition of a mature phenotype and type I IFN secretion. The data therefore demonstrate that DCs can respond to HSV exposure directly through recognition of viral envelope structures. In the context of natural HSV infection, the coupling of viral entry to the activation of DC signaling pathways is likely to be counterbalanced by viral disruption of DC maturation. However, the parallel release of type I IFN may result in paracrine activation so that the DCs are nonetheless able to mount an adaptive immune response. PMID- 15356162 TI - Activation of signaling lymphocytic activation molecule triggers a signaling cascade that enhances Th1 responses in human intracellular infection. AB - T cell production of IFN-gamma contributes to host defense against infection by intracellular pathogens, including mycobacteria. Lepromatous leprosy, the disseminated form of infection caused by Mycobacterium leprae, is characterized by loss of cellular response against the pathogen and diminished Th1 cytokine production. Relieving bacterial burden in Ag-unresponsive patients might be achieved through alternative receptors that stimulate IFN-gamma production. We have previously shown that ligation of signaling lymphocytic activation molecule (SLAM) enhances IFN-gamma in mycobacterial infection; therefore, we investigated molecular pathways leading from SLAM activation to IFN-gamma production in human leprosy. The expression of the SLAM-associated protein (an inhibitory factor for IFN-gamma induction) on M. leprae-stimulated cells from leprosy patients was inversely correlated to IFN-gamma production. However, SLAM ligation or exposure of cells from lepromatous patients to a proinflammatory microenvironment down regulated SLAM-associated protein expression. Moreover, SLAM activation induced a sequence of signaling proteins, including activation of the NF-kappaB complex, phosphorylation of Stat1, and induction of T-bet expression, resulting in the promotion of IFN-gamma production, a pathway that remains quiescent in response to Ag in lepromatous patients. Therefore, our findings reveal a cascade of molecular events during signaling through SLAM in leprosy that cooperate to induce IFN-gamma production and strongly suggest that SLAM might be a focal point for therapeutic modulation of T cell cytokine responses in diseases characterized by dysfunctional Th2 responses. PMID- 15356163 TI - Platelet-derived thrombospondin-1 is necessary for the vitamin D-binding protein (Gc-globulin) to function as a chemotactic cofactor for C5a. AB - The chemotactic activity of C5a and C5a des Arg can be enhanced significantly by the vitamin D-binding protein (DBP), also known as Gc-globulin. DBP is a multifunctional 56-kDa plasma protein that binds and transports several diverse ligands. The objective of this study was to investigate the mechanisms by which DBP functions as a chemotactic cofactor for C5a using neutrophils and U937 cells transfected with the C5aR (U937-C5aR cells). The results demonstrate that U937 C5aR cells show C5a chemotactic enhancement only to DBP in serum, but, unlike mature neutrophils, this cell line cannot respond to DBP in plasma or to purified DBP. Analysis by SDS-PAGE and isoelectric focusing revealed no structural difference between DBP in serum compared with DBP in plasma. However, plasma supplemented with either serum, DBP-depleted serum, or activated platelet releasate provides a required factor and permits DBP to function as a chemotactic cofactor for C5a. Fractionation of activated platelet releasate revealed that the additional factor possessed the properties of thrombospondin-1 (TSP-1). Finally, purified TSP-1 alone could reproduce the effect of serum or platelet releasate, whereas Abs to TSP-1 could block these effects. These results provide clear evidence that TSP-1 is needed for DBP to function as a chemotactic cofactor for C5a. PMID- 15356164 TI - The essential role of the intestinal microbiota in facilitating acute inflammatory responses. AB - The restoration of blood flow, i.e., reperfusion, is the treatment of choice to save viable tissue following acute ischemia of a vascular territory. Nevertheless, reperfusion can be accompanied by significant inflammatory events that limit the beneficial effects of blood flow restoration. To evaluate the potential role of the intestinal microbiota in facilitating the development of tissue injury and systemic inflammation, germ-free and conventional mice were compared in their ability to respond to ischemia and reperfusion injury. In conventional mice, there was marked local (intestine) and remote (lung) edema formation, neutrophil influx, hemorrhage, and production of TNF-alpha, KC, MIP-2, and MCP-1. Moreover, there was an increase in the concentration of serum TNF alpha and 100% lethality. In germ-free mice, there was no local, remote, or systemic inflammatory response or lethality after intestinal ischemia and reperfusion and, in contrast to conventional mice, germ-free animals produced greater amounts of IL-10. Similar results were obtained after administration of LPS, i.e., little production of TNF-alpha or lethality and production of IL-10 after LPS in germ-free mice. Blockade of IL-10 with Abs induced marked inflammation and lethality in germ-free mice after ischemia and reperfusion or LPS administration, demonstrating that the ability of these mice to produce IL-10 was largely responsible for their "no inflammation" phenotype. This was consistent with the prevention of reperfusion-associated injury by the exogenous administration of IL-10 to conventional mice. Thus, the lack of intestinal microbiota is accompanied by a state of active IL-10-mediated inflammatory hyporesponsiveness. PMID- 15356165 TI - Reduction of nitric oxide synthase 2 expression by distamycin A improves survival from endotoxemia. AB - NO synthase 2 (NOS2) plays an important role in endotoxemia through overproduction of NO. Distamycin A (Dist A) belongs to a class of drugs termed minor-groove DNA binders, which can inhibit transcription factor binding to AT rich regions of DNA. We and others have previously shown that AT-rich regions of DNA surrounding transcription factor binding sites in the NOS2 promoter are critical for NOS2 induction by inflammatory stimuli in vitro. Therefore, we hypothesized that Dist A would attenuate NOS2 up-regulation in vivo during endotoxemia and improve animal survival. C57BL/6 wild-type (WT) mice treated with Dist A and LPS (endotoxin) showed significantly improved survival compared with animals treated with LPS alone. In contrast, LPS-treated C57BL/6 NOS2-deficient (NOS2-/-) mice did not benefit from the protective effect of Dist A on mortality from endotoxemia. Treatment with Dist A resulted in protection from hypotension in LPS-treated WT mice, but not in NOS2-/- mice. Furthermore, LPS-induced NOS2 expression was attenuated in vivo (WT murine tissues) and in vitro (primary peritoneal and RAW 264.7 murine macrophages) with addition of Dist A. Dist A selectively decreased IFN regulatory factor-1 DNA binding in the enhancer region of the NOS2 promoter, and this IFN regulatory factor-1 site is critical for the effect of Dist A in attenuating LPS induction of NOS2. Our data point to a novel approach in modulating NOS2 expression in vivo during endotoxemia and suggest the potential for alternative treatment approaches for critical illness. PMID- 15356166 TI - Regulation of human polymorphonuclear leukocytes functions by the neuropeptide pituitary adenylate cyclase-activating polypeptide after activation of MAPKs. AB - Anti-inflammatory activities of pituitary adenylate cyclase-activating protein (PACAP) are mediated in part through specific effects on lymphocytes and macrophages. This study shows that in human polymorphonuclear neutrophils (PMNs), PACAP acts as a proinflammatory molecule. In PMNs, vaso-intestinal peptide/PACAP receptor 1 (VPAC-1) was the only receptor found to be expressed by RT-PCR. Using VPAC-1 Ab, we found that VPAC-1 mRNA was translated into proteins. In PMNs, PACAP increases cAMP, inositol triphosphate metabolites, and calcium. It activates two of the three members of the MAPK superfamily, the ERK and the stress-activated MAPK p38. U73122, an inhibitor of phospholipase C (PLC), inhibits PACAP-induced ERK activation, whereas p38 MAPK phosphorylation was unaffected. Using specific pharmalogical inhibitors of ERK (PD098059) and p38 MAPK (SB203580), we found that PACAP-mediated calcium increase was ERK and PLC dependent and p38 independent. PACAP primes fMLP-associated calcium increase; it also primes fMLP activation of the respiratory burst as well as elastase release, these last two processes being ERK and PLC dependent and p38 MAPK independent. PACAP also increases membrane expression of CD11b and release of lactoferrin and metallo proteinase-9 (MMP-9). These effects were PLC dependent (CD 11b, lactoferrin, MMP-9), ERK dependent (CD 11b, lactoferrin, MMP-9), and p38 dependent (CD11b, lactoferrin). We conclude that PACAP is a direct PMN activator as well as an effective PMN priming agent that requires PLC, ERK, and p38 MAPK activities. PMID- 15356167 TI - Cigarette smoke inhibits lipopolysaccharide-induced production of inflammatory cytokines by suppressing the activation of activator protein-1 in bronchial epithelial cells. AB - Chronic smoking is characterized by immunosuppressive changes in the airways, leading to chronic colonization with bacteria, which in turn may contribute to the chronic obstructive pulmonary disease. The mechanisms causing this immunosuppression, however, are poorly characterized. This study evaluated whether cigarette smoke can inhibit endotoxin (LPS)-induced inflammatory cytokine production in bronchial epithelial cells and, if so, what the mechanisms are behind this effect. Pretreatment with cigarette smoke extract (CSE) concentration dependently inhibited the LPS-induced GM-CSF and IL-8 protein release, which was accompanied by decreased expression of mRNA in human bronchial epithelial cells (Beas-2B). The increase of neutrophil chemotaxis induced by conditioned medium from LPS-treated Beas-2B cells was also suppressed by CSE. In addition, the activity of LPS-induced transcription factor AP-1, but not NF-kappaB, was down regulated by CSE. Notably, at the concentrations used, CSE had no effect on number or viability of Beas-2B cells. These data indicate that cigarette smoke possesses immunosuppressive properties by down-regulating the bacterial pathogen induced neutrophil-mobilizing cytokine production via suppression of AP-1 activation in the airways. Hence, this study suggests a novel mechanism by which cigarette smoke may contribute to chronic colonization and chronic obstructive pulmonary disease in smokers. PMID- 15356168 TI - Modulation of renal disease in MRL/lpr mice by suberoylanilide hydroxamic acid. AB - Epigenetic regulation of gene expression is involved in the development of many diseases. Histone acetylation is a posttranslational modification of the nucleosomal histone tails that is regulated by the balance of histone deacetylases and histone acetyltransferases. Alterations in the balance of histone acetylation have been shown to cause aberrant expression of genes that are a hallmark of many diseases, including systemic lupus erythematosus. In this study, we determined whether suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor: 1) inhibits inflammatory mediator production in vitro and 2) modulates lupus progression in vivo. Mesangial cells isolated from 10-wk-old MRL/lpr mice were stimulated with LPS/IFN-gamma and incubated with SAHA. TNF alpha, IL-6, NO, and inducible NO synthase expression were inhibited by SAHA. We then treated MRL/lpr mice with daily injections of SAHA from age 10 to 20 wk. The animals treated with SAHA had decreased spleen size and a concomitant decrease in CD4-CD8- (double-negative) T cells compared with controls. Serum autoantibody levels and glomerular IgG and C3 deposition in SAHA-treated mice were similar to controls. In contrast, proteinuria and pathologic renal disease were significantly inhibited in the mice receiving SAHA. These data indicate that SAHA blocks mesangial cell inflammatory mediator production in vitro and disease progression in vivo in MRL/lpr mice. PMID- 15356169 TI - Proinflammatory cytokines induce proteinase 3 as membrane-bound and secretory forms in human oral epithelial cells and antibodies to proteinase 3 activate the cells through protease-activated receptor-2. AB - Anti-neutrophil cytoplasmic Abs targeting proteinase 3 (PR3) have been detected in relation to a wide range of inflammatory conditions such as periodontitis, and interaction of anti-PR3 Abs with endothelial and epithelial cells provokes cell activation, although the underlying mechanism has been unclear. The present study showed that human oral epithelial cells expressed PR3 mRNA after treatment with proinflammatory cytokines such as IL-1alpha, TNF-alpha, IFN-alpha, IFN-beta, and IFN-gamma. A 29-kDa PR3 was expressed on the cell surface and released into culture supernatants by the cells upon stimulation with these cytokines. The membrane and supernatant fractions of oral epithelial cells exhibited enzymatic activity, which was inhibited by serine proteinase inhibitors, but not by a cysteine proteinase inhibitor or secretory leukocyte protease inhibitor. Addition of anti-PR3 Abs to cytokine-primed oral epithelial cells in culture induced remarkable secretion of IL-8 and monocyte chemoattractant protein 1 and aggregation of PR3 on the cells. RNA interference targeted to protease-activated receptor-2 mRNA and intracellular Ca2+ mobilization assays revealed that anti-PR3 Abs activated the epithelial cells through protease-activated receptor-2, a family of G protein-coupled receptors. The anti-PR3 Ab-mediated cell activation was completely abolished by RNA interference targeted to PR3 mRNA and by inhibition of phospholipase C and NF-kappaB. Immunohistochemistry showed that inflamed oral epithelium actually expresses PR3 protein. These results suggest that oral epithelial cells express functional PR3 in the inflamed sites and respond to anti-PR3 Abs detected in diseased sera, and that these mechanisms may actively participate in the inflammatory process, including periodontitis. PMID- 15356170 TI - Renal expression of the C3a receptor and functional responses of primary human proximal tubular epithelial cells. AB - Although complement activation and deposition have been associated with a variety of glomerulopathies, the pathogenic mechanisms by which complement directly mediates renal injury remain to be fully elucidated. Renal parenchymal tissues express a limited repertoire of receptors that directly bind activated complement proteins. We report the renal expression of the receptor for the C3 cleavage product C3a, a member of the anaphylatoxin family. C3aR is highly expressed in normal human and murine kidney, as demonstrated by immunohistochemistry and in situ hybridization. Its distribution is limited to epithelial cells only, as glomerular endothelial and mesangial cells showed no evidence of C3aR expression. The C3aR is also expressed by primary renal proximal tubular epithelial cells in vitro as demonstrated by FACS, Western blot, and RT-PCR. In vitro C3aR is functional in terms of its capacity to bind 125I-labeled C3a and generate inositol triphosphate. Finally, using microarray analysis, four novel genes were identified and confirmed as transcriptionally regulated by C3aR activation in proximal tubular cells. These studies define a new pathway by which complement activation may directly modulate the renal response to immunologic injury. PMID- 15356171 TI - IL-8 is a key chemokine regulating neutrophil recruitment in a new mouse model of Shigella-induced colitis. AB - The lack of a mouse model of acute rectocolitis mimicking human bacillary dysentery in the presence of invasive Shigella is a major handicap to study the pathogenesis of the disease and to develop a Shigella vaccine. The inability of the mouse intestinal mucosa to elicit an inflammatory infiltrate composed primarily of polymorphonuclear leukocytes (PMN) may be due to a defect in epithelial invasion, in the sensing of invading bacteria, or in the effector mechanisms that recruit the PMN infiltrate. We demonstrate that the BALB/cJ mouse colonic epithelium not only can be invaded by Shigella, but also elicits an inflammatory infiltrate that, however, lacks PMN. This observation points to a major defect of mice in effector mechanisms, particularly the lack of expression of the CXC chemokine, IL-8. Indeed, this work demonstrates that the delivery of recombinant human IL-8, together with Shigella infection of the colonic epithelial surface, causes an acute colitis characterized by a strong PMN infiltrate that, by all criteria, including transcription profiles of key mediators of the innate/inflammatory response and histopathological lesions, mimics bacillary dysentery. This is a major step forward in the development of a murine model of bacillary dysentery. PMID- 15356172 TI - Andrographolide attenuates inflammation by inhibition of NF-kappa B activation through covalent modification of reduced cysteine 62 of p50. AB - NF-kappaB is a central transcriptional factor and a pleiotropic regulator of many genes involved in immunological responses. During the screening of a plant extract library of traditional Chinese herbal medicines, we found that NF-kappaB activity was potently inhibited by andrographolide (Andro), an abundant component of the plant Andrographis that has been commonly used as a folk remedy for alleviation of inflammatory disorders in Asia for millennia. Mechanistically, it formed a covalent adduct with reduced cysteine (62) of p50, thus blocking the binding of NF-kappaB oligonucleotide to nuclear proteins. Andro suppressed the activation of NF-kappaB in stimulated endothelial cells, which reduced the expression of cell adhesion molecule E-selectin and prevented E-selectin-mediated leukocyte adhesion under flow. It also abrogated the cytokine- and endotoxin induced peritoneal deposition of neutrophils, attenuated septic shock, and prevented allergic lung inflammation in vivo. Notably, it had no suppressive effect on IkappaBalpha degradation, p50 and p65 nuclear translocation, or cell growth rates. Our results thus reveal a unique pharmacological mechanism of Andro's protective anti-inflammatory actions. PMID- 15356173 TI - CD137-deficient mice have reduced NK/NKT cell numbers and function, are resistant to lipopolysaccharide-induced shock syndromes, and have lower IL-4 responses. AB - CD137, a member of the TNF superfamily, is involved in T cell and NK cell activation and cytokine production. To establish its in vivo role in systems dependent on NK and NKT cells, we studied the response of CD137-/- mice to LPS induced shock, tumor killing, and their IL-4-controlled Th2 responses. In both high and low dose shock models, all the CD137-deficient mice, but none of the wild-type BALB/c mice, survived. After injection of LPS/2-amino-2-deoxy-D galactose (D-gal), CD137-/- mice had reduced serum cytokine levels and substantially impaired liver IFN-gamma and TNF-alpha mRNA levels. Phenotypic analysis of mononuclear cells revealed fewer NK and NKT cells in the CD137-/- mice. The knockout mice did not generate a rapid IL-4 response after systemic T cell activation, or effective Ag-specific Th2 responses. In addition, both in vitro and in vivo NK-specific cytolytic activities were reduced. These findings suggest that CD137-directed NK/NKT cells play an important role in the inflammatory response leading to the production of proinflammatory cytokines, LPS induced septic shock, and tumor killing, as well as IL-4-dependent Th2 responses. PMID- 15356174 TI - Accelerated ischemia/reperfusion-induced injury in autoimmunity-prone mice. AB - Natural Abs have been implicated in initiating mesenteric ischemia/reperfusion (I/R)-induced tissue injury. Autoantibodies have affinity and self-Ag recognition patterns similar to natural Abs. We considered that autoimmunity-prone mice that express high titers of autoantibodies should have enhanced I/R-induced injury. Five-month-old B6.MRL/lpr mice displayed accelerated and enhanced intestinal I/R induced damage compared with 2-mo-old B6.MRL/lpr and age-matched C57BL/6 mice. Similarly, older autoimmune mice had accelerated remote organ (lung) damage. Infusion of serum IgG derived from 5-mo-old but not 2-mo-old B6.MRL/lpr into I/R resistant Rag-1-/- mice rendered them susceptible to local and remote organ injury. Injection of monoclonal IgG anti-DNA and anti-histone Abs into Rag-1-/- mice effectively reconstituted tissue injury. These data show that like natural Abs, autoantibodies, such as anti-dsDNA and anti-histone Abs, can instigate I/R injury and suggest that they are involved in the development of tissue damage in patients with systemic lupus erythematosus. PMID- 15356175 TI - Complement mediates the binding of HIV to erythrocytes. AB - A fraction of HIV is associated with erythrocytes even when the virus becomes undetectable in plasma under antiretroviral therapy. The aim of the present work was to further characterize this association in vitro. We developed an in vitro model to study the factors involved in the adherence of HIV-1 to erythrocytes. Radiolabeled HIV-1 (HIV) and preformed HIV-1/anti-HIV immune complexes (HIV-IC) were opsonized in various human sera, purified using sucrose density gradient ultracentrifugation, and incubated with human erythrocytes. We observed that, when opsonized in normal human serum, not only HIV-IC, but also HIV, bound to erythrocytes, although the adherence of HIV was lower than that of HIV-IC. The adherence was abolished when the complement system was blocked, but was maintained in hypogammaglobulinemic sera. Complement-deficient sera indicated that both pathways of complement were important for optimal adherence. No adherence was seen in C1q-deficient serum, and the adherence of HIV was reduced when the alternative pathway was blocked using anti-factor D Abs. The adherence could be inhibited by an mAb against complement receptor 1. At supraphysiological concentrations, purified C1q mediated the binding of a small fraction of HIV and HIV-IC to erythrocytes. In conclusion, HIV-IC bound to erythrocytes as other types of IC do when exposed to complement. Of particular interest was that HIV alone bound also to erythrocytes in a complement/complement receptor 1-dependent manner. Thus, erythrocytes may not only deliver HIV-IC to organs susceptible to infection, but free HIV as well. This may play a crucial role in the progression of the primary infection. PMID- 15356176 TI - Interactions between LTP- and LTD-inducing stimulation in the sensorimotor cortex of the awake freely moving rat. AB - Bidirectional modifications in synaptic efficacy are central components in models of cortical learning and memory. More recently, the regulation of synaptic plasticity according to the history of synaptic activation, termed "metaplasticity," has become a focus of research on the physiology of memory. Here we explore such interactions between long-term potentiation (LTP) and long term depression (LTD) in the chronically prepared rat. The effects of successive high- and low-frequency stimulation were examined in sensorimotor cortex in the adult, freely moving rat. High-frequency (300 Hz) stimulation (HFS) applied to the white matter was used to induce LTP, and prolonged, low-frequency (1 Hz) stimulation (LFS) was used to induce either depotentiation or LTD. Combined stimulation (HFS/LFS or LFS/HFS) during the induction phase attenuated potentiation effects only if the LFS followed the HFS. LTD induced by LFS alone was expressed as a reduction in the amplitude of both short- and long-latency field potential components, whereas depotentiation was primarily expressed as a decrease in the amplitude of the potentiated long-latency component. In other experiments, LTP (or LTD) was induced to asymptotic levels before applying LFS (or HFS). LFS caused depotentiation of the late component but had no measurable effect on the early component. HFS reversed previously induced LTD, but the potentiation decayed more rapidly than usual. LTP and LTD therefore modulate each other in the awake, behaving rat. PMID- 15356177 TI - Contribution of afferent feedback to the soleus muscle activity during human locomotion. AB - During the stance phase of the human step cycle, the ankle undergoes a natural dorsiflexion that stretches the soleus muscle. The afferent feedback resulting from this stretch enhances the locomotor drive. In this study a robotic actuator was used to slightly enhance or reduce the natural ankle dorsiflexion, in essence, mimicking the small variations in the ankle dorsiflexion movement that take place during the stance phase of the step cycle. The soleus (SOL) and tibialis anterior EMG were analyzed in response to the ankle trajectory modifications. The dorsiflexion enhancements and reductions generated gradual increments and decrements, respectively, in the ongoing SOL EMG. We exercised care to ensure that the imposed ankle movements were too slow to elicit distinct burst-like stretch reflex responses that have been investigated previously. The increased SOL EMG after the dorsiflexion enhancements was reduced when the group Ia afferents were blocked with peripheral ischemia at the thigh, and during high frequency Achilles tendon vibration. However, neither ischemia nor tendon vibration affected the decrements in the SOL EMG during the dorsiflexion reductions. These findings give evidence of the contribution of afferent feedback to the SOL activity in an ongoing basis during the stance phase. The results suggest that mainly feedback from the group Ia pathways is responsible for the increments in the SOL EMG during the dorsiflexion enhancements. However, the decrements in the SOL activity might be mediated by different afferent mechanisms. PMID- 15356178 TI - Regulation of synaptic inputs to paraventricular-spinal output neurons by alpha2 adrenergic receptors. AB - Neurons in the paraventricular nucleus (PVN) that project to the brain stem and spinal cord are important for autonomic regulation. The excitability of preautonomic PVN neurons is controlled by the noradrenergic input from the brain stem. In this study, we determined the role of alpha(2) adrenergic receptors in the regulation of excitatory and inhibitory synaptic inputs to spinally projecting PVN neurons. Excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) were recorded using whole cell voltage-clamp techniques on PVN neurons labeled by a retrograde fluorescence tracer injected into the thoracic spinal cord of rats. Bath application of 5-20 muM clonidine, an alpha(2) receptor agonist, significantly reduced the amplitude of evoked GABAergic IPSCs in a dose dependent manner. Also, 10 microM clonidine significantly decreased the frequency (from 2.68 +/- 0.41 to 1.22 +/- 0.40 Hz) but not the amplitude of miniature IPSCs (mIPSCs), and this effect was blocked by the alpha(2) receptor antagonist yohimbine. Furthermore, clonidine increased the paired-pulse ratio of evoked IPSCs from 1.25 +/- 0.05 to 1.61 +/- 0.08 (P < 0.05). On the other hand, clonidine had little effect on evoked glutamatergic EPSCs, mEPSCs, and the paired pulse ratio of evoked EPSCs in most labeled cells examined. Additionally, immunofluorescence labeling revealed that the alpha(2A) receptor and GABA immunoreactivities were co-localized in close apposition to labeled PVN neurons. Collectively, these data suggest that stimulation of alpha(2) adrenergic receptors primarily attenuates GABAergic inputs to PVN output neurons to the spinal cord. The presynaptic alpha(2) receptors function as heteroreceptors to modulate synaptic GABA release and contribute to the hypothalamic regulation of sympathetic outflow. PMID- 15356179 TI - Contralateral White Noise Selectively Changes Right Human Auditory Cortex Activity Caused by a FM-Direction Task. AB - Animal and human studies suggest that directional categorization of frequency modulated (FM) tones (rising vs. falling) is a function of the right auditory cortex (AC). To investigate this hemispheric specialization in more detail, we analyzed both the binaural and monaural representation of FM tones and the influence of contralateral white noise on the processing of FM tone direction. In two fMRI-experiments, FM tones with varied direction, center-frequencies, and duration were presented binaurally or monaurally without contralateral white noise (experiment 1) and with contralateral white noise (experiment 2) while the subjects had to perform the same directional categorization task. In experiment 1, contralateral FM tones led to strongest activation, binaural FM tones to intermediate, and ipsilateral FM tones to weakest activation in each AC. This is in accordance with binaural response properties of neurons in animal AC. In experiment 2, contralateral white noise had no significant effect on the activation of left AC by FM tones, whereas in right AC, it led to a significant increase in activation for ipsilateral FM tones. This result provides further support for the critical role of right AC for directional categorization of FM tones, which for ipsilateral input has to be processed in competition to the excitatory input of white noise via the direct contralateral pathway. PMID- 15356180 TI - Synaptic depression in conjunction with A-current channels promote phase constancy in a rhythmic network. AB - In many central pattern generators, pairs of neurons maintain an approximately fixed phase despite large changes in the frequency. The mechanisms underlying phase maintenance are not clear. Previous theoretical work suggested that inhibitory synapses that show short-term depression could play a critical role in this respect. In this work we examine how the interaction between synaptic depression and the kinetics of a transient potassium (A-like) current could be advantageous for phase constancy in a rhythmic network. To demonstrate the mechanism in the context of a realistic central pattern generator, we constructed a detailed model of the crustacean pyloric circuit. The frequency of the rhythm was modified by changing the level of a ligand-activated current in one of the pyloric neurons. We examined how the time difference of firing activities between two selected neurons in this circuit is affected by synaptic depression, A current, and a combination of the two. We tuned the parameters of the model such that with synaptic depression alone, or A-current alone, phase was not maintained between these two neurons. However, when these two components came together, they acted synergistically to maintain the phase across a wide range of cycle periods. This suggests that synaptic depression may be necessary to allow an A-current to delay a postsynaptic neuron in a frequency-dependent manner, such that phase invariance is ensured. PMID- 15356181 TI - Different inhibitory inputs onto neostriatal projection neurons as revealed by field stimulation. AB - This work investigated if diverse properties could be ascribed to evoked inhibitory postsynaptic currents (IPSCs) recorded on rat neostriatal neurons when field stimulation was delivered at two different locations: the globus pallidus (GP) and the neostriatum (NS). Previous work stated that stimulation in the GP could antidromically excite projection axons from medium spiny neurons. This maneuver would predominantly activate the inhibitory synapses that interconnect spiny cells. In contrast, intrastriatal stimulation would preferentially activate inhibitory synapses provided by interneurons. This study shows that, in fact, intensity-amplitude experiments are able to reveal different properties for IPSCs evoked from these two locations (GP and NS). In addition, while all IPSCs evoked from the GP were always sensitive to omega-conotoxin GVIA (Ca(V2.2)2.2 or N channel blocker), one-half of the inhibition evoked from the NS exhibited little sensitivity to omega-conotoxin GVIA. Characteristically, all omega-conotoxin GVIA insensitive IPSCs exhibited strong paired pulse depression, whereas omega conotoxin GVIA-sensitive IPSCs evoked from either the GP or the NS could exhibit short-time depression or facilitation. omega-Agatoxin TK (Ca(V2.1)2.1+ or P/Q channel blocker) blocked IPSCs evoked from both locations. Therefore 1) distinct inhibitory inputs onto projection neostriatal cells can be differentially stimulated with field electrodes; 2) N-type Ca2+ channels are not equally expressed in inhibitory terminals activated in the NS; and 3) synapses that interconnect spiny neurons use both N- and P/Q-type Ca2+ channels. PMID- 15356182 TI - Cerebellar involvement in anticipating the consequences of self-produced actions during bimanual movements. AB - Anticipatory postural adjustments (APA) during bimanual actions can be observed when participants hold an object in one hand and then lift it with the other hand. The postural force used to hold the object is reduced in anticipation of unloading, indicating an accurate prediction of the change in load. We examined patients with unilateral or bilateral cerebellar damage as well as two individuals lacking the corpus callosum on the bimanual unloading task. The acallosal patients showed an intact APA, suggesting subcortical integration of motor signals for anticipatory adjustments during bimanual actions. Contrary to the hypothesis that the cerebellum is critical for predicting and compensating for the consequences of our actions, we found that the well-learned APA in this task was largely intact in cerebellar patients. However, cerebellar damage abolished short-term adaptation of the APA, and the patients were unable to acquire an APA in a similar but previously untrained situation. These results indicate that while over-learned anticipatory adjustments are preserved after cerebellar lesions, adaptation of this response and the acquisition of a novel coordination requires the cerebellum ipsilateral to the postural hand. Furthermore, this structure appears to be essential for the accurate timing of previously learned behaviors. The patients with cerebellar damage showed poorly timed adjustments with the APA beginning earlier than in healthy participants. PMID- 15356183 TI - A point process framework for relating neural spiking activity to spiking history, neural ensemble, and extrinsic covariate effects. AB - Multiple factors simultaneously affect the spiking activity of individual neurons. Determining the effects and relative importance of these factors is a challenging problem in neurophysiology. We propose a statistical framework based on the point process likelihood function to relate a neuron's spiking probability to three typical covariates: the neuron's own spiking history, concurrent ensemble activity, and extrinsic covariates such as stimuli or behavior. The framework uses parametric models of the conditional intensity function to define a neuron's spiking probability in terms of the covariates. The discrete time likelihood function for point processes is used to carry out model fitting and model analysis. We show that, by modeling the logarithm of the conditional intensity function as a linear combination of functions of the covariates, the discrete time point process likelihood function is readily analyzed in the generalized linear model (GLM) framework. We illustrate our approach for both GLM and non-GLM likelihood functions using simulated data and multivariate single unit activity data simultaneously recorded from the motor cortex of a monkey performing a visuomotor pursuit-tracking task. The point process framework provides a flexible, computationally efficient approach for maximum likelihood estimation, goodness-of-fit assessment, residual analysis, model selection, and neural decoding. The framework thus allows for the formulation and analysis of point process models of neural spiking activity that readily capture the simultaneous effects of multiple covariates and enables the assessment of their relative importance. PMID- 15356184 TI - Integration of target and effector information in human posterior parietal cortex for the planning of action. AB - Recently, using event-related functional MRI (fMRI), we located a bilateral region in the human posterior parietal cortex (retIPS) that topographically represents and updates targets for saccades and pointing movements in eye centered coordinates. To generate movements, this spatial information must be integrated with the selected effector. We now tested whether the activation in retIPS is dependent on the hand selected. Using 4-T fMRI, we compared the activation produced by movements, using either eyes or the left or right hand, to targets presented either leftward or rightward of central fixation. The majority of the regions activated during saccades were also activated during pointing movements, including occipital, posterior parietal, and premotor cortex. The topographic retIPS region was activated more strongly for saccades than for pointing. The activation associated with pointing was significantly greater when pointing with the unseen hand to targets ipsilateral to the hand. For example, although there was activation in the left retIPS when pointing to targets on the right with the left hand, the activation was significantly greater when using the right hand. The mirror symmetric effect was observed in the right retIPS. Similar hand preferences were observed in a nearby anterior occipital region. This effector specificity is consistent with previous clinical and behavioral studies showing that each hand is more effective in directing movements to targets in ipsilateral visual space. We conclude that not only do these regions code target location, but they also appear to integrate target selection with effector selection. PMID- 15356185 TI - Contribution of persistent sodium currents to spike-frequency adaptation in rat hypoglossal motoneurons. AB - In response to constant current inputs, the firing rates of motoneurons typically show a continuous decline over time. The biophysical mechanisms underlying this process, called spike-frequency adaptation, are not well understood. Spike frequency adaptation normally exhibits a rapid initial phase, followed by a slow, later phase that continues throughout the duration of firing. One possible mechanism mediating the later phase might be a reduction in the persistent sodium current (I(NaP)) that has been shown to diminish the capacity of cortical pyramidal neurons and spinal motoneurons to sustain repetitive firing. In this study, we used the anticonvulsant phenytoin to reduce the I(NaP) of juvenile rat hypoglossal motoneurons recorded in brain stem slices, and we examined the consequences of a reduction in I(NaP) on the magnitude and time course of spike frequency adaptation. Adding phenytoin to the bathing solution (> or =50 microM) generally produced a marked reduction in the persistent inward currents (PICs) recorded at the soma in response to slow, voltage-clamp triangular ramp commands (-70 to 0 mV and back). However, the same concentrations of phenytoin appeared to have no significant effect on spike-frequency adaptation even though the phenytoin often augmented the reduction in action potential amplitude that occurs during repetitive firing. The surprising finding that the reduction of a source of sustained inward current had no appreciable effect on the pattern of spike generation suggests that several types of membrane channels must act cooperatively to insure that these motoneurons can generate the sustained repetitive firing required for long-lasting motor behaviors. PMID- 15356186 TI - Angiotensin II excites paraventricular nucleus neurons that innervate the rostral ventrolateral medulla: an in vitro patch-clamp study in brain slices. AB - Neurons of the hypothalamic paraventricular nucleus (PVN) are key controllers of sympathetic nerve activity and receive input from angiotensin II (ANG II) containing neurons in the forebrain. This study determined the effect of ANG II on PVN neurons that innervate in the rostral ventrolateral medulla (RVLM)-a brain stem site critical for maintaining sympathetic outflow and arterial pressure. Using an in vitro brain slice preparation, whole cell patch-clamp recordings were made from PVN neurons retrogradely labeled from the ipsilateral RVLM of rats. Of 71 neurons tested, 62 (87%) responded to ANG II. In current-clamp mode, bath applied ANG II (2 muM) significantly (P < 0.05) depolarized membrane potential from -58.5 +/- 2.5 to -54.5 +/- 2.0 mV and increased the frequency of action potential discharge from 0.7 +/- 0.3 to 2.8 +/- 0.8 Hz (n = 4). Local application of ANG II by low-pressure ejection from a glass pipette (2 pmol, 0.4 nl, 5 s) also elicited rapid and reproducible excitation in 17 of 20 cells. In this group, membrane potential depolarization averaged 21.5 +/- 4.1 mV, and spike activity increased from 0.7 +/- 0.4 to 21.3 +/- 3.3 Hz. In voltage-clamp mode, 41 of 47 neurons responded to pressure-ejected ANG II with a dose-dependent inward current that averaged -54.7 +/- 3.9 pA at a maximally effective dose of 2.0 pmol. Blockade of ANG II AT1 receptors significantly reduced discharge (P < 0.001, n = 5), depolarization (P < 0.05, n = 3), and inward current (P < 0.01, n = 11) responses to locally applied ANG II. In six of six cells tested, membrane input conductance increased (P < 0.001) during local application of ANG II (2 pmol), suggesting influx of cations. The ANG II current reversed polarity at +2.2 +/- 2.2 mV (n = 9) and was blocked (P < 0.01) by bath perfusion with gadolinium (Gd(3+), 100 muM, n = 8), suggesting that ANG II activates membrane channels that are nonselectively permeable to cations. These findings indicate that ANG II excites PVN neurons that innervate the ipsilateral RVLM by a mechanism that depends on activation of AT1 receptors and gating of one or more classes of ion channels that result in a mixed cation current. PMID- 15356187 TI - Adenosine postsynaptically modulates supraoptic neuronal excitability. AB - Effects of adenosine on the excitability of supraoptic nucleus neurons were investigated in whole cell patch-clamp experiments conducted in horizontal slices of rat hypothalamus. Adenosine (10-100 muM) inhibited all neurons tested by reducing or abolishing spontaneous or evoked discharge. Large hyperpolarizations were seen, averaging -6.08 +/- 0.83 mV below resting membrane potential, and action potential durations were significantly reduced by 134 +/- 41 mus in the presence of 100 muM adenosine. The A(1) receptor antagonist 8-cyclopentyl-1,3 dipropylxanthine (DPCPX, 1 muM) blocked these effects, whereas the A(1) agonists N(6)-cyclopentyladenosine (CPA) and N(6)-cyclohexyladenosine (CHA) mimicked the actions of adenosine. A(2) receptor contributions to excitability were assessed by application of an A(2) agonist, carboxamidoadenosine (CPCA). This resulted in membrane depolarizations (3.56 +/- 0.65 mV) and maintenance of firing. The presence of endogenous adenosine in the slice was revealed by both the application of the adenosine uptake inhibitor dilazep (1-100 muM), which resulted in a strong inhibition of firing activity, and the application of DPCPX, which induced firing in cells silenced by negative current injection. We tested for postsynaptic actions of adenosine by blocking G protein activation via GDP-beta-S infusion into recorded neurons. Under these conditions, the adenosinergic inhibition of firing and reduction of spike duration were blocked, suggesting the effects were mediated by postsynaptic adenosine receptors. That the effects on excitability could be due to direct activation of adenosine A(1) receptors on supraoptic neurons was further explored immunocytochemically via the co-labeling of magnocellular neurons with polyclonal antibodies raised against the A(1) receptors. It is concluded that adenosine, acting at postsynaptic A(1) receptors, exhibits a powerful inhibitory influence on supraoptic magnocellular activity and is an important endogenous regulator of magnocellular neuroendocrine function. PMID- 15356188 TI - Generation of reactive oxygen species in the reaction catalyzed by alpha ketoglutarate dehydrogenase. AB - Alpha-ketoglutarate dehydrogenase (alpha-KGDH), a key enzyme in the Krebs' cycle, is a crucial early target of oxidative stress (Tretter and Adam-Vizi, 2000). The present study demonstrates that alpha-KGDH is able to generate H(2)O(2) and, thus, could also be a source of reactive oxygen species (ROS) in mitochondria. Isolated alpha-KGDH with coenzyme A (HS-CoA) and thiamine pyrophosphate started to produce H(2)O(2) after addition of alpha-ketoglutarate in the absence of nicotinamide adenine dinucleotide-oxidized (NAD(+)). NAD(+), which proved to be a powerful inhibitor of alpha-KGDH-mediated H(2)O(2) formation, switched the H(2)O(2) forming mode of the enzyme to the catalytic [nicotinamide adenine dinucleotide-reduced (NADH) forming] mode. In contrast, NADH stimulated H(2)O(2) formation by alpha-KGDH, and for this, neither alpha-ketoglutarate nor HS-CoA were required. When all of the substrates and cofactors of the enzyme were present, the NADH/NAD(+) ratio determined the rate of H(2)O(2) production. The higher the NADH/NAD(+) ratio the higher the rate of H(2)O(2) production. H(2)O(2) production as well as the catalytic function of the enzyme was activated by Ca(2+). In synaptosomes, using alpha-ketoglutarate as respiratory substrate, the rate of H(2)O(2) production increased by 2.5-fold, and aconitase activity decreased, indicating that alpha-KGDH can generate H(2)O(2) in in situ mitochondria. Given the NADH/NAD(+) ratio as a key regulator of H(2)O(2) production by alpha-KGDH, it is suggested that production of ROS could be significant not only in the respiratory chain but also in the Krebs' cycle when oxidation of NADH is impaired. Thus alpha-KGDH is not only a target of ROS but could significantly contribute to generation of oxidative stress in the mitochondria. PMID- 15356189 TI - Mitochondrial alpha-ketoglutarate dehydrogenase complex generates reactive oxygen species. AB - Mitochondria-produced reactive oxygen species (ROS) are thought to contribute to cell death caused by a multitude of pathological conditions. The molecular sites of mitochondrial ROS production are not well established but are generally thought to be located in complex I and complex III of the electron transport chain. We measured H(2)O(2) production, respiration, and NADPH reduction level in rat brain mitochondria oxidizing a variety of respiratory substrates. Under conditions of maximum respiration induced with either ADP or carbonyl cyanide p trifluoromethoxyphenylhydrazone,alpha-ketoglutarate supported the highest rate of H(2)O(2) production. In the absence of ADP or in the presence of rotenone, H(2)O(2) production rates correlated with the reduction level of mitochondrial NADPH with various substrates, with the exception of alpha-ketoglutarate. Isolated mitochondrial alpha-ketoglutarate dehydrogenase (KGDHC) and pyruvate dehydrogenase (PDHC) complexes produced superoxide and H(2)O(2). NAD(+) inhibited ROS production by the isolated enzymes and by permeabilized mitochondria. We also measured H(2)O(2) production by brain mitochondria isolated from heterozygous knock-out mice deficient in dihydrolipoyl dehydrogenase (Dld). Although this enzyme is a part of both KGDHC and PDHC, there was greater impairment of KGDHC activity in Dld-deficient mitochondria. These mitochondria also produced significantly less H(2)O(2) than mitochondria isolated from their littermate wild type mice. The data strongly indicate that KGDHC is a primary site of ROS production in normally functioning mitochondria. PMID- 15356190 TI - Ceramidase regulates synaptic vesicle exocytosis and trafficking. AB - A screen for Drosophila synaptic dysfunction mutants identified slug-a-bed (slab). The slab gene encodes ceramidase, a central enzyme in sphingolipid metabolism and regulation. Sphingolipids are major constituents of lipid rafts, membrane domains with roles in vesicle trafficking, and signaling pathways. Null slab mutants arrest as fully developed embryos with severely reduced movement. The SLAB protein is widely expressed in different tissues but enriched in neurons at all stages of development. Targeted neuronal expression of slab rescues mutant lethality, demonstrating the essential neuronal function of the protein. C(5) ceramide applied to living preparations is rapidly accumulated at neuromuscular junction (NMJ) synapses dependent on the SLAB expression level, indicating that synaptic sphingolipid trafficking and distribution is regulated by SLAB function. Evoked synaptic currents at slab mutant NMJs are reduced by 50-70%, whereas postsynaptic glutamate-gated currents are normal, demonstrating a specific presynaptic impairment. Hypertonic saline-evoked synaptic vesicle fusion is similarly impaired by 50-70%, demonstrating a loss of readily releasable vesicles. In addition, FM1-43 dye uptake is reduced in slab mutant presynaptic terminals, indicating a smaller cycling vesicle pool. Ultrastructural analyses of mutants reveal a normal vesicle distribution clustered and docked at active zones, but fewer vesicles in reserve regions, and a twofold to threefold increased incidence of vesicles linked together and tethered at the plasma membrane. These results indicate that SLAB ceramidase function controls presynaptic terminal sphingolipid composition to regulate vesicle fusion and trafficking, and thus the strength and reliability of synaptic transmission. PMID- 15356191 TI - Ipsilateral actions of feline corticospinal tract neurons on limb motoneurons. AB - Contralateral pyramidal tract (PT) neurons arising in the primary motor cortex are the major route through which volitional limb movements are controlled. However, the contralateral hemiparesis that follows PT neuron injury on one side may be counteracted by ipsilateral of actions of PT neurons from the undamaged side. To investigate the spinal relays through which PT neurons may influence ipsilateral motoneurons, we analyzed the synaptic actions evoked by stimulation of the ipsilateral pyramid on hindlimb motoneurons after transecting the descending fibers of the contralateral PT at a low thoracic level. The results show that ipsilateral PT neurons can affect limb motoneurons trisynaptically by activating contralaterally descending reticulospinal neurons, which in turn activate spinal commissural interneurons that project back across to motoneurons ipsilateral to the stimulated pyramidal tract. Stimulation of the pyramids alone did not evoke synaptic actions in motoneurons but potently facilitated disynaptic EPSPs and IPSPs evoked by stimulation of reticulospinal tract fibers in the medial longitudinal fascicle. In parallel with this double-crossed pathway, corticospinal neurons could also evoke ipsilateral actions via ipsilateral descending reticulospinal tract fibers, acting through ipsilaterally located spinal interneurons. Because the actions mediated by commissural interneurons were found to be stronger than those of ipsilateral premotor interneurons, the study leads to the conclusion that ipsilateral actions of corticospinal neurons via commissural interneurons may provide a better opportunity for recovery of function in hemiparesis produced by corticospinal tract injury. PMID- 15356192 TI - Developmental regulation of nicotinic synapses on cochlear inner hair cells. AB - In the mature cochlea, inner hair cells (IHCs) transduce acoustic signals into receptor potentials, communicating to the brain by synaptic contacts with afferent fibers. Before the onset of hearing, a transient efferent innervation is found on IHCs, mediated by a nicotinic cholinergic receptor that may contain both alpha9 and alpha10 subunits. Calcium influx through that receptor activates calcium-dependent (SK2-containing) potassium channels. This inhibitory synapse is thought to disappear after the onset of hearing [after postnatal day 12 (P12)]. We documented this developmental transition using whole-cell recordings from IHCs in apical turns of the rat organ of Corti. Acetylcholine elicited ionic currents in 88-100% of IHCs between P3 and P14, but in only 1 of 11 IHCs at P16-P22. Potassium depolarization of efferent terminals caused IPSCs in 67% of IHCs at P3, in 100% at P7-P9, in 93% at P10-P12, but in only 40% at P13-P14 and in none of the IHCs tested between P16 and P22. Earlier work had shown by in situ hybridization that alpha9 mRNA is expressed in adult IHCs but that alpha10 mRNA disappears after the onset of hearing. In the present study, antibodies to alpha10 and to the associated calcium-dependent (SK2) potassium channel showed a similar developmental loss. The correlated expression of these gene products with functional innervation suggests that Alpha10 and SK2, but not Alpha9, are regulated by synaptic activity. Furthermore, this developmental knock-out of alpha10, but not alpha9, supports the hypothesis that functional nicotinic acetylcholine receptors in hair cells are heteromers containing both these subunits. PMID- 15356193 TI - Differential roles of NR2A and NR2B-containing NMDA receptors in cortical long term potentiation and long-term depression. AB - It is widely believed that long-term depression (LTD) and its counterpart, long term potentiation (LTP), involve mechanisms that are crucial for learning and memory. However, LTD is difficult to induce in adult cortex for reasons that are not known. Here we show that LTD can be readily induced in adult cortex by the activation of NMDA receptors (NMDARs), after inhibition of glutamate uptake. Interestingly there is no need to activate synaptic NMDARs to induce this LTD, suggesting that LTD is triggered primarily by extrasynaptic NMDA receptors. We also find that de novo LTD requires the activation of NR2B-containing NMDAR, whereas LTP requires activation of NR2A-containing NMDARs. Surprisingly another form of LTD, depotentiation, requires activation of NR2A-containing NMDARs. Therefore, NMDARs with different synaptic locations and subunit compositions are involved in various forms of synaptic plasticity in adult cortex. PMID- 15356194 TI - Lasting blood-brain barrier disruption induces epileptic focus in the rat somatosensory cortex. AB - Perturbations in the integrity of the blood-brain barrier have been reported in both humans and animals under numerous pathological conditions. Although the blood-brain barrier prevents the penetration of many blood constituents into the brain extracellular space, the effect of such perturbations on the brain function and their roles in the pathogenesis of cortical diseases are unknown. In this study we established a model for focal disruption of the blood-brain barrier in the rat cortex by direct application of bile salts. Exposure of the cerebral cortex in vivo to bile salts resulted in long-lasting extravasation of serum albumin to the brain extracellular space and was associated with a prominent activation of astrocytes with no inflammatory response or marked cell loss. Using electrophysiological recordings in brain slices we found that a focus of epileptiform discharges developed within 4-7 d after treatment and could be recorded up to 49 d postoperatively in >60% of slices from treated animals but only rarely (10%) in sham-operated controls. Epileptiform activity involved both glutamatergic and GABAergic neurotransmission. Epileptiform activity was also induced by direct cortical application of native serum, denatured serum, or albumin-containing solution. In contrast, perfusion with serum-adapted electrolyte solution did not induce abnormal activity, thereby suggesting that the exposure of the serum-devoid brain environment to serum proteins underlies epileptogenesis in the blood-brain barrier-disrupted cortex. Although many neuropathologies entail a compromised blood-brain barrier, this is the first direct evidence that it may have a role in the pathogenesis of focal cortical epilepsy, a common neurological disease. PMID- 15356195 TI - L1.1 is involved in spinal cord regeneration in adult zebrafish. AB - Adult zebrafish, in contrast to mammals, regrow axons descending from the brainstem after spinal cord transection. L1.1, a homolog of the mammalian recognition molecule L1, is upregulated by brainstem neurons during axon regrowth. However, its functional relevance for regeneration is unclear. Here, we show with a novel morpholino-based approach that reducing L1.1 protein expression leads to impaired locomotor recovery as well as reduced regrowth and synapse formation of axons of supraspinal origin after spinal cord transection. This indicates that L1.1 contributes to successful regrowth of axons from the brainstem and locomotor recovery after spinal cord transection in adult zebrafish. PMID- 15356196 TI - Hypothermia-associated loss of dendritic spines. AB - Mechanisms of synaptic plasticity in CNS circuits are commonly investigated using in vitro preparations such as brain slices or slice culture. During their preparation, slices are exposed to low temperatures, and electrophysiological measurements are sometimes made below physiological temperature. Because dendritic spines, which occur at the majority of excitatory synapses, are morphologically plastic, we investigated the influence of reduced temperature on their morphology and plasticity using live cell imaging of hippocampal slices from transgenic mice expressing a green fluorescent protein-based neuronal surface marker and electron microscopy of adult brain slices. Our data show that dendritic spines are highly sensitive to reduced temperature with rapid loss of actin-based motility followed at longer times by reversible loss of the entire spine structure. Thus, reduced temperature significantly affects synaptic morphology, which is in turn known to influence several key aspects of synaptic transmission. Evidence that hypothermia potentiates anesthesia and is associated with spine loss in hibernating animals further suggests that spine morphology may have a widespread influence on brain function. PMID- 15356197 TI - Functional properties of motoneurons derived from mouse embryonic stem cells. AB - The capacity of embryonic stem (ES) cells to form functional motoneurons (MNs) and appropriate connections with muscle was investigated in vitro. ES cells were obtained from a transgenic mouse line in which the gene for enhanced green fluorescent protein (eGFP) is expressed under the control of the promotor of the MN specific homeobox gene Hb9. ES cells were exposed to retinoic acid (RA) and sonic hedgehog agonist (Hh-Ag1.3) to stimulate differentiation into MNs marked by expression of eGFP and the cholinergic transmitter synthetic enzyme choline acetyltransferase. Whole-cell patch-clamp recordings were made from eGFP-labeled cells to investigate the development of functional characteristics of MNs. In voltage-clamp mode, currents, including EPSCs, were recorded in response to exogenous applications of GABA, glycine, and glutamate. EGFP-labeled neurons also express voltage-activated ion channels including fast-inactivating Na(+) channels, delayed rectifier and I(A)-type K(+) channels, and Ca(2+) channels. Current-clamp recordings demonstrated that eGFP-positive neurons generate repetitive trains of action potentials and that l-type Ca(2+) channels mediate sustained depolarizations. When cocultured with a muscle cell line, clustering of acetylcholine receptors on muscle fibers adjacent to developing axons was seen. Intracellular recordings of muscle fibers adjacent to eGFP-positive axons revealed endplate potentials that increased in amplitude and frequency after glutamate application and were sensitive to TTX and curare. In summary, our findings demonstrate that MNs derived from ES cells develop appropriate transmitter receptors, intrinsic properties necessary for appropriate patterns of action potential firing and functional synapses with muscle fibers. PMID- 15356198 TI - Chronic ethanol induces synaptic but not extrasynaptic targeting of NMDA receptors. AB - The development of ethanol tolerance and dependence reflects neuroadaptive changes in response to continuous depression in synaptic activity. The present study used confocal imaging and electrophysiology procedures to assess the effects of prolonged ethanol exposure on NMDA receptor trafficking in cultures of hippocampal neurons. Neurons exposed to 50 mm ethanol for 4 d showed an increase in the colocalization of NMDA receptor type 1 (NR1) clusters with the presynaptic marker protein synapsin. This was accompanied by significant increases in the size and density of these synapsin-associated clusters with no change observed in nonsynapsin-associated NR1 clusters. Similar effects were observed with NR2B clustering after chronic ethanol exposure. The increase in synaptic NMDA receptor clustering was prevented by addition of a protein kinase A inhibitor or by coexposure to a low concentration of NMDA and was reversed when ethanol was removed from the cultures. No changes were observed in the synaptic content, cluster size, or density of AMPA receptors after ethanol exposure. Electrophysiological measurements on ethanol-treated neurons revealed a similar enhancement in synaptic NMDA currents with no change in AMPA-mediated events. After isolation of extrasynaptic NMDA receptors by MK801 (+)-5-methyl-10,11 dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate (/) trapping, whole-cell responses to NMDA were not different between control and ethanol-treated neurons These observations demonstrate that neuroadaptive changes in NMDA receptors in response to prolonged ethanol exposure occur through activity-dependent processes that regulate their synaptic targeting and localization. PMID- 15356199 TI - Loss of flight and associated neuronal rhythmicity in inositol 1,4,5 trisphosphate receptor mutants of Drosophila. AB - Coordinated flight in winged insects requires rhythmic activity of the underlying neural circuit. Here, we show that Drosophila mutants for the inositol 1,4,5 trisphosphate (InsP(3)) receptor gene (itpr) are flightless. Electrophysiological recordings from thoracic indirect flight muscles show increased spontaneous firing accompanied by a loss of rhythmic flight activity patterns normally generated in response to a gentle puff of air. In contrast, climbing speed, the jump response, and electrical properties of the giant fiber pathway are normal, indicating that general motor coordination and neuronal excitability are much less sensitive to itpr mutations. All mutant phenotypes are rescued by expression of an itpr(+) transgene in serotonin and dopamine neurons. Pharmacological and immunohistochemical experiments support the idea that the InsP(3) receptor functions to modulate flight specifically through serotonergic interneurons. InsP(3) receptor action appears to be important for normal development of the flight circuit and its central pattern generator. PMID- 15356200 TI - The c-Jun N-terminal protein kinase signaling pathway mediates Bax activation and subsequent neuronal apoptosis through interaction with Bim after transient focal cerebral ischemia. AB - The c-Jun N-terminal protein kinase (JNK) signaling pathway is implicated in neuronal apoptosis. The mechanism by which activated JNK induces neuronal apoptosis is strongly linked to mitochondrial apoptogenic proteins, although the molecular machinery downstream of JNK has not been precisely elucidated. Our study examined the relevance of proapoptotic Bcl-2 family members in JNK-mediated apoptosis after transient focal cerebral ischemia (tFCI), which, when induced by 60 min of middle cerebral artery (MCA) occlusion, elevated levels of JNK activity and phospho-JNK in the MCA territory. Phospho-JNK was primarily expressed in neurons and colocalized with terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL)-positive cells. Inhibition of JNK activity by anthra[1,9-cd]pyrazol-6(2H)-one (SP600125), a selective JNK inhibitor, protected neurons from ischemia-induced apoptosis detected by TUNEL staining and an apoptotic-related DNA fragmentation assay. SP600125 blocked translocation of the cell death effector Bax from the cytosol to the mitochondria after tFCI. BimL (Bim long) was induced and phosphorylated parallel to JNK activity. Coimmunoprecipitation studies consistently revealed increased interaction of JNK with BimL, as well as BimL with Bax, after tFCI. SP600125 blocked these interactions at a dose that significantly inhibited JNK-induced neuronal apoptosis. These results suggest that the JNK signaling pathway is involved in ischemia-induced neuronal apoptosis by stimulation, at least in part, of Bax translocation to the mitochondria, in which BimL is likely regulated by JNK as a downstream substrate for transmission of apoptotic signals to Bax. PMID- 15356201 TI - Uptake and release of norepinephrine by serotonergic terminals in norepinephrine transporter knock-out mice: implications for the action of selective serotonin reuptake inhibitors. AB - Our aim was to investigate the functional properties of the noradrenergic system in genetically modified mice lacking the norepinephrine transporter (NET). We measured the uptake and release of [(3)H]norepinephrine ([(3)H]NE) from hippocampal and cortical slices of NET(-/-) knock-out (KO) and NET(+/+) wild-type (WT) mice and investigated the presynaptic alpha2-adenoceptor-mediated modulation of NE release in vitro and in vivo. The [(3)H]NE uptake was reduced to 12.6% (hippocampus) and 33.5% (frontal cortex) of WT control in KO mice. The neuronal component of this residual uptake was decreased by 79.4 and 100%, respectively, when a selective serotonin reuptake inhibitor (SSRI) citalopram was present during the loading. The more preserved neuronal release of [(3)H]NE (hippocampus, 28.1%; frontal cortex, 74.4%; compared with WT) almost completely disappeared in both regions (94.1 and 95.3% decrease compared with KO, respectively) in the presence of citalopram, suggesting that [(3)H]NE was taken up and released by serotonergic varicosities. This was further supported by the finding that the release of [(3)H]NE from hippocampal slices of KO mice was not modulated by the alpha2-adrenoceptor antagonist 7,8-(methylenedioxy)-14-alpha-hydroxyalloberbane HCl, whereas the endogenous release of NE measured by microdialysis was even more efficiently enhanced by this drug in NET-deficient mice. These experiments indicate that serotonergic varicosities can accumulate and release NE as a result of the heterologous uptake of transmitters. Because the diffusion of NE may be spatially limited by serotonin transporters, the SSRIs, despite their selectivity, might enhance not only serotonergic but also noradrenergic neurotransmission, which might contribute to their antidepressant action. PMID- 15356202 TI - Early N-terminal changes and caspase-6 cleavage of tau in Alzheimer's disease. AB - Alzheimer's disease (AD) is a progressive amnestic dementia that involves post translational hyperphosphorylation, enzymatic cleavage, and conformational alterations of the microtubule-associated protein tau. The truncation state of tau influences many of its pathologic characteristics, including its ability to assume AD-related conformations and to assemble into filaments. Cleavage also appears to be an important marker in AD progression. Although C-terminal truncation of tau at D421 has recently been attributed to the apoptotic enzyme caspase-3, N-terminal processing of the protein remains mostly uncharacterized. Here, we report immunohistochemical staining in a cohort of 35 cases ranging from noncognitively impaired to early AD with a panel of three N-terminal anti-tau antibodies: Tau-12, 5A6, and 9G3-pY18. Of these three, the phosphorylation independent epitope of 5A6 was the earliest to emerge in the pathological lesions of tau, followed by the appearance of the Tau-12 epitope. The unmasking of the Tau-12 epitope in more mature 5A6-positive tangles was not correlated with tau phosphorylation at tyrosine 18 (9G3-pY18). Still, later in the course of tangle evolution, the extreme N terminus of tau was lost, correlating temporally with the appearance of a C-terminal caspase-truncated epitope lacking residues 422 441. In addition, caspase-6 cleaved the N terminus of tau in vitro, preventing immunoreactivity with both Tau-12 and 5A6. Mass spectrometry confirmed that the in vitro caspase-6 truncation site is D13, a semicanonical and hitherto undescribed caspase cleavage site in tau. Collectively, these results suggest a role for caspase-6 and N-terminal truncation of tau during neurofibrillary tangle evolution and the progression of Alzheimer's disease. PMID- 15356203 TI - KChIPs and Kv4 alpha subunits as integral components of A-type potassium channels in mammalian brain. AB - Voltage-gated potassium (Kv) channels from the Kv4, or Shal-related, gene family underlie a major component of the A-type potassium current in mammalian central neurons. We recently identified a family of calcium-binding proteins, termed KChIPs (Kv channel interacting proteins), that bind to the cytoplasmic N termini of Kv4 family alpha subunits and modulate their surface density, inactivation kinetics, and rate of recovery from inactivation (An et al., 2000). Here, we used single and double-label immunohistochemistry, together with circumscribed lesions and coimmunoprecipitation analyses, to examine the regional and subcellular distribution of KChIPs1-4 and Kv4 family alpha subunits in adult rat brain. Immunohistochemical staining using KChIP-specific monoclonal antibodies revealed that the KChIP polypeptides are concentrated in neuronal somata and dendrites where their cellular and subcellular distribution overlaps, in an isoform specific manner, with that of Kv4.2 and Kv4.3. For example, immunoreactivity for KChIP1 and Kv4.3 is concentrated in the somata and dendrites of hippocampal, striatal, and neocortical interneurons. Immunoreactivity for KChIP2, KChIP4, and Kv4.2 is concentrated in the apical and basal dendrites of hippocampal and neocortical pyramidal cells. Double-label immunofluorescence labeling revealed that throughout the forebrain, KChIP2 and KChIP4 are frequently colocalized with Kv4.2, whereas in cortical, hippocampal, and striatal interneurons, KChIP1 is frequently colocalized with Kv4.3. Coimmunoprecipitation analyses confirmed that all KChIPs coassociate with Kv4 alpha subunits in brain membranes, indicating that KChIPs 1-4 are integral components of native A-type Kv channel complexes and are likely to play a major role as modulators of somatodendritic excitability. PMID- 15356204 TI - Comparison of IPSCs evoked by spiny and fast-spiking neurons in the neostriatum. AB - Most neurons in the neostriatum are GABAergic spiny projection neurons with extensive local axon collaterals innervating principally other spiny projection neurons. The other source of GABAergic inputs to spiny neurons derives from a small number of interneurons, of which the best characterized are the parvalbumin containing, fast-spiking interneurons. Spiny neuron collateral inhibition was not demonstrated until recently, because the IPSPs recorded at the soma are surprisingly small. In contrast, interneuronal inhibition was readily detected, comprising much larger IPSPs. Here, we report the application of quantal analysis and compartmental modeling to compare and contrast IPSCs in spiny neurons originating from axon collaterals and interneurons. The results indicate that individual release sites at spiny and interneuron synapses have similar quantal sizes and baseline release probabilities. Interneuronal unitary IPSCs are several times larger because of their proximal location on the neuron and because they have a larger number of transmitter release sites. Despite the small amount of current they can deliver to the soma, spiny cell collateral synapses had moderately high baseline release probabilities (0.5-0.9), suggesting that they are not weak because of some form of depression or modulation. The size of unitary collateral synaptic currents increased monotonically during development. These results argue against models of competitive inhibition in neostriatum, including those in which competitive inhibition is transiently effective during development and learning, and suggest a different role for the spiny cell axon collaterals. PMID- 15356205 TI - The upregulated expression of sonic hedgehog in motor neurons after rat facial nerve axotomy. AB - Nerve injury leads to the induction of a large number of genes to repair the damage and to restore synaptic transmission. We have attempted to identify molecules whose mRNA expression is altered in response to facial nerve axotomy. Here we report that facial nerve axotomy upregulates Sonic hedgehog (Shh) and its receptor Smoothened (Smo) in facial motor neurons of adult rats, whereas facial nerve axotomy does not upregulate mRNA of Shh or Smo in neonatal rats. We tested whether overexpression of Shh in facial motor neurons of axotomized neonatal rats may promote neuronal survival. Adenovirus-mediated overexpression of Shh, but not that of beta-galactosidase, transiently rescues axotomy-induced neuronal cell death for 3-5 d after axotomy. Finally, the pharmacological inhibitor of Shh signaling, cyclopamine, induces motor neuron death in adult rats after axotomy. These results suggest that Shh plays a regulatory role in nerve injury. PMID- 15356206 TI - Chloride accumulation in mammalian olfactory sensory neurons. AB - The generation of an excitatory receptor current in mammalian olfactory sensory neurons (OSNs) involves the sequential activation of two distinct types of ion channels: cAMP-gated Ca(2+)-permeable cation channels and Ca(2+)-gated Cl(-) channels, which conduct a depolarizing Cl(-) efflux. This unusual transduction mechanism requires an outward-directed driving force for Cl(-), established by active accumulation of Cl(-) within the lumen of the sensory cilia. We used two photon fluorescence lifetime imaging microscopy of the Cl(-)-sensitive dye 6 methoxy-quinolyl acetoethyl ester to measure the intracellular Cl(-) concentration in dendritic knobs of OSNs from mice and rats. We found a uniform intracellular Cl(-) concentration in the range of 40-50 mm, which is indicative of active Cl(-) accumulation. Functional assays and PCR experiments revealed that NKCC1-mediated Cl(-) uptake through the apical membrane counteracts Cl(-) depletion in the sensory cilia, and thus maintains the responsiveness of OSNs to odor stimulation. To permit Cl(-) accumulation, OSNs avoid the "chloride switch": they do not express KCC2, the main Cl(-) extrusion cotransporter operating in neurons of the adult CNS. Cl(-) accumulation provides OSNs with the driving force for the depolarizing Cl(-) current that is the basis of the low-noise receptor current in these neurons. PMID- 15356207 TI - Age-related functional changes of prefrontal cortex in long-term memory: a repetitive transcranial magnetic stimulation study. AB - Neuroimaging findings suggest that the lateralization of prefrontal cortex activation associated with episodic memory performance is reduced by aging. It is still a matter of debate whether this loss of asymmetry during encoding and retrieval reflects compensatory mechanisms or de-differentiation processes. We addressed this issue by the transient interference produced by repetitive transcranial magnetic stimulation (rTMS), which directly assesses causal relationships between performance and stimulated regions. We compared the effects of rTMS (a rapid-rate train occurring simultaneously to the presentation of memoranda) applied to the left or right dorsolateral prefrontal cortex (DLPFC) on visuospatial recognition memory in 66 healthy subjects divided in two classes of age (<45 and >50 years). In young subjects, rTMS of the right DLPFC interfered with retrieval more than left DLPFC stimulation. The asymmetry of the effect progressively vanished with aging, as indicated by bilateral interference effects on recognition performance. Conversely, the predominance of left DLPFC effect during encoding was not abolished in elders, thus probing its causal role for encoding along the life span. Findings confirm that the neural correlates of retrieval modify along aging, suggesting that the bilateral engagement of the DLPFC has a compensatory role on the elders' episodic memory performance. PMID- 15356208 TI - Genetically decreased spinal cord copper concentration prolongs life in a transgenic mouse model of amyotrophic lateral sclerosis. AB - Mutations in the Cu/Zn superoxide dismutase (SOD1) gene cause familial amyotrophic lateral sclerosis (FALS) by gain of an aberrant function that is not yet well understood. The role of Cu(2+) in mediating the toxicity of mutant SOD1 has been earnestly contested. We tested the in vivo effects of genetically induced copper deprivation on the ALS phenotype of transgenic mice expressing G86R mutant mouse SOD1, a protein that fails to incorporate Cu(2+) in its active site. Genetically copper-deficient SOD1(G86R) transgenic mice were produced by mating SOD1(G86R) males to female carriers of the X-linked mottled/brindled (Mobr) mutation. We found that the Mobr allele causes a severe ( approximately 60%) depletion of spinal cord copper levels; however, despite the burden of double genetic lesions, it lengthens the lives of SOD1(G86R) transgenic mice by 9%. These findings provide evidence supporting a role for copper in the pathogenesis of FALS linked to SOD1 mutations. PMID- 15356209 TI - The neuropeptide pigment-dispersing factor coordinates pacemaker interactions in the Drosophila circadian system. AB - In Drosophila, the neuropeptide pigment-dispersing factor (PDF) is required to maintain behavioral rhythms under constant conditions. To understand how PDF exerts its influence, we performed time-series immunostainings for the PERIOD protein in normal and pdf mutant flies over 9 d of constant conditions. Without pdf, pacemaker neurons that normally express PDF maintained two markers of rhythms: that of PERIOD nuclear translocation and its protein staining intensity. As a group, however, they displayed a gradual dispersion in their phasing of nuclear translocation. A separate group of non-PDF circadian pacemakers also maintained PERIOD nuclear translocation rhythms without pdf but exhibited altered phase and amplitude of PERIOD staining intensity. Therefore, pdf is not required to maintain circadian protein oscillations under constant conditions; however, it is required to coordinate the phase and amplitude of such rhythms among the diverse pacemakers. These observations begin to outline the hierarchy of circadian pacemaker circuitry in the Drosophila brain. PMID- 15356210 TI - Brain-derived neurotrophic factor plays a critical role in contextual fear conditioning. AB - In this study, brain-derived neurotrophic factor (BDNF) heterozygous knock-outs were tested on fear conditioning, and their wild-type littermates were used as controls. Results showed that BDNF(+/-) mice are impaired in contextual learning, whereas tone learning remains intact. Because BDNF is involved in synaptic transmission and contextual learning is hippocampal dependent, we hypothesized that this deficit is attributable to abnormal BDNF-modulated synaptic plasticity in the hippocampus. A "gain-of-function" experiment was performed next by infusing recombinant BDNF protein into the hippocampal formation to investigate whether this deficit can be rescued. Infusion of BDNF protein into the hippocampus appeared to partially restore contextual fear learning of BDNF(+/-) mice. In conclusion, the present study suggests that BDNF plays a critical role in fear conditioning. Loss of one copy of the BDNF gene leads to impairment of contextual fear learning in BDNF(+/-). This deficit can be partially rescued by infusing BDNF protein into the hippocampus. Other brain regions interacting with the hippocampus in the context conditioned stimulus pathway, for example, the amygdala, may also require normal BDNF expression levels to fully rescue this impairment. PMID- 15356211 TI - Attentional modulation of motion integration of individual neurons in the middle temporal visual area. AB - We examined how spatially directed attention affected the integration of motion in neurons of the middle temporal (MT) area of visual cortex. We recorded from single MT neurons while monkeys performed a motion detection task under two attentional states. Using 0% coherent random dot motion, we estimated the optimal linear transfer function (or kernel) between the global motion and the neuronal response. This linear kernel filtered the random dot motion across direction, speed, and time. Slightly less than one-half of the neurons produced reasonably well defined kernels that also tended to account for both the directional selectivity and responses to coherent motion of different strengths. This subpopulation of cells had faster, more transient, and more robust responses to visual stimuli than neurons with kernels that did not contain well defined regions of integration. For those neurons that had large attentional modulation and produced well defined kernels, we found attention scaled the temporal profile of the transfer function with no appreciable shift in time or change in shape. Thus, for MT neurons described by a linear transfer function, attention produced a multiplicative scaling of the temporal integration window. PMID- 15356212 TI - Extreme stretch growth of integrated axons. AB - Large animals can undergo enormous growth during development, suggesting that axons in nerves and white matter tracts rapidly expand as well. Because integrated axons have no growth cones to extend from, it has been postulated that mechanical forces may stimulate axon elongation matching the growth of the animal. However, this distinct form of rapid and sustained growth of integrated axons has never been demonstrated. Here, we used a microstepper motor system to evaluate the effects of escalating rates of stretch on integrated axon tracts over days to weeks in culture. We found that axon tracts could be stretch grown at rates of 8 mm/d and reach lengths of 10 cm without disconnection. Despite dynamic and long-term elongation, stretched axons increased in caliber by 35%, while the morphology and density of cytoskeletal constituents and organelles were maintained. These data provide the first evidence that mechanical stimuli can induce extreme "stretch growth" of integrated axon tracts, far exceeding any previously observed limits of axon growth. PMID- 15356213 TI - Glucagon-like peptide-1 limits myocardial stunning following brief coronary occlusion and reperfusion in conscious canines. AB - We have recently demonstrated the benefits of glucagon-like peptide-1 (GLP-1) in enhancing regional and global myocardial function after reperfusion in the clinical setting of acute myocardial infarction. We hypothesized that GLP-1 facilitates recovery from myocardial stunning after an ischemic event. To investigate this, we administered GLP-1 (1.5 pmol/kg/min) to six dogs undergoing 10-min occlusion of the left circumflex coronary artery, followed by 24-h reperfusion. We compared the responses of coronary blood flow and regional thickening of the posterior wall with a group of eight vehicle-treated dogs undergoing the same occlusion-reperfusion protocol. Although recovery of coronary blood flow was identical, regional wall motion recovery occurred significantly ((*)p < 0.05) earlier (92 +/- 4 versus 57 +/- 5%(*) at 15 min) and was complete in the GLP-1-treated dogs, whereas residual contractile dysfunction persisted in the control group (99 +/- 4 versus 78 +/- 3%(*) at 24 h). This phenomenon was independent of changes in systemic hemodynamics or global systolic function. However, isovolumic left ventricular relaxation improved significantly in GLP-1 treated dogs. GLP-1 caused an insulinotropic effect, but no hypoglycemia. We conclude that GLP-1 enhances recovery from ischemic myocardial stunning after successful reperfusion. PMID- 15356214 TI - Peroxisome proliferation-activated receptor (PPAR)gamma is not necessary for synthetic PPARgamma agonist inhibition of inducible nitric-oxide synthase and nitric oxide. AB - Peroxisome proliferation-activated receptor (PPAR)gamma agonists inhibit inducible nitric-oxide synthase (iNOS), tumor necrosis factor-alpha, and interleukin-6. Because of these effects, synthetic PPARgamma agonists, including thiazolidinediones, are being studied for their impact on inflammatory disease. The anti-inflammatory concentrations of synthetic PPARgamma agonists range from 10 to 50 microM, whereas their binding affinity for PPARgamma is in the nanomolar range. The specificity of synthetic PPARgamma agonists for PPARgamma at the concentrations necessary for anti-inflammatory effects is thus in question. We report that PPARgamma is not necessary for the inhibition of iNOS by synthetic PPARgamma agonists. RAW 264.7 macrophages possess little PPARgamma, yet lipopolysaccharide (LPS)/interferon (IFN)gamma-induced iNOS was inhibited by synthetic PPARgamma agonists at 20 microM. Endogenous PPARgamma was inhibited by the transfection of a dominant-negative PPARgamma construct into murine mesangial cells. In the transfected cells, synthetic PPARgamma agonists inhibited iNOS production at 10 microM, similar to nontransfected cells. Using cells from PPARgamma Cre/lox conditional knockout mice, baseline and LPS/IFNgamma-induced nitric oxide levels were higher in macrophages lacking PPARgamma versus controls. However, synthetic PPARgamma agonists inhibited iNOS at 10 microM in the PPARgamma-deficient cells, similar to macrophages from wild-type mice. These results indicate that PPARgamma is not necessary for inhibition of iNOS expression by synthetic PPARgamma agonists at concentrations over 10 microM. Intrinsic PPARgamma function, in the absence of synthetic agonists, however, may play a role in inflammatory modulation. PMID- 15356215 TI - C75 [4-methylene-2-octyl-5-oxo-tetrahydro-furan-3-carboxylic acid] activates carnitine palmitoyltransferase-1 in isolated mitochondria and intact cells without displacement of bound malonyl CoA. AB - Carnitine palmitoyltransferase 1beta (CPT-1beta) is a key regulator of the beta oxidation of long-chain fatty acids in skeletal muscle and therefore a potential therapeutic target for diseases associated with defects in lipid metabolism such as obesity and type 2 diabetes. C75 [4-methylene-2-octyl-5-oxo-tetrahydro-furan-3 carboxylic acid] is an alpha-methylene-butyrolactone that has been characterized as both an inhibitor of fatty acid synthase and more recently, an activator of CPT-1 (Thupari et al., 2002). Using human CPT-1beta expressed in the yeast Pichia pastoris, we demonstrate that C75 can activate the skeletal muscle isoform of CPT 1 and overcome inactivation of the enzyme by malonyl CoA, an important physiological repressor of CPT-1, and the malonyl CoA mimetic Ro25-0187 [{5-[2 (naphthalen-2-yloxy)-ethoxy]-thiophen-2-yl}-oxo-acetic acid]. We also show that C75 can activate CPT-1 in intact hepatocytes to levels similar to those achieved with inhibition of acetyl-CoA carboxylase, the enzyme that produces malonyl CoA. Finally, we demonstrate that concentrations of C75 sufficient for activation of CPT-1 do not displace bound malonyl CoA. We conclude that CPT-1 is an activator of human CPT-1beta and other CPT-1 isoforms but that it does not activate CPT-1 through antagonism of malonyl CoA binding. PMID- 15356216 TI - Development of the first ultra-potent "capsaicinoid" agonist at transient receptor potential vanilloid type 1 (TRPV1) channels and its therapeutic potential. AB - Olvanil (N-9-Z-octadecenoyl-vanillamide) is an agonist of transient receptor potential vanilloid type 1 (TRPV1) channels that lack the pungency of capsaicin and was developed as an oral analgesic. Vanillamides are unmatched in terms of structural simplicity, straightforward synthesis, and safety compared with the more powerful TRPV1 agonists, like the structurally complex phorboid compound resiniferatoxin. We have modified the fatty acyl chain of olvanil to obtain ultra potent analogs. The insertion of a hydroxyl group at C-12 yielded a compound named rinvanil, after ricinoleic acid, significantly less potent than olvanil (EC(50) = 6 versus 0.7 nM), but more versatile in terms of structural modifications because of the presence of an additional functional group. Acetylation and phenylacetylation of rinvanil re-established and dramatically enhanced, respectively, its potency at hTRPV1. With a two-digit picomolar EC(50) (90 pM), phenylacetylrinvanil (PhAR, IDN5890) is the most potent vanillamide ever described with potency comparable with that of resiniferatoxin (EC(50), 11 pM). Benzoyl- and phenylpropionylrinvanil were as potent and less potent than PhAR, respectively, whereas configurational inversion to ent-PhAR and cyclopropanation (but not hydrogenation or epoxidation) of the double bond were tolerated. Finally, iodination of the aromatic hydroxyl caused a dramatic switch in functional activity, generating compounds that behaved as TRPV1 antagonists rather than agonists. Since the potency of PhAR was maintained in rat dorsal root ganglion neurons and, particularly, in the rat urinary bladder, this compound was investigated in an in vivo rat model of urinary incontinence and proved as effective as resiniferatoxin at reducing bladder detrusor overactivity. PMID- 15356217 TI - Heterologous expression of human {alpha}6{beta}4{beta}3{alpha}5 nicotinic acetylcholine receptors: binding properties consistent with their natural expression require quaternary subunit assembly including the {alpha}5 subunit. AB - Heterologous expression and lesioning studies were conducted to identify possible subunit assembly partners in nicotinic acetylcholine receptors (nAChR) containing alpha6 subunits (alpha6(*) nAChR). SH-EP1 human epithelial cells were transfected with the requisite subunits to achieve stable expression of human alpha6beta2, alpha6beta4, alpha6beta2beta3, alpha6beta4beta3, or alpha6beta4beta3alpha5 nAChR. Cells expressing subunits needed to form alpha6beta4beta3alpha5 nAChR exhibited saturable [(3)H]epibatidine binding (K(d) = 95.9 +/- 8.3 pM and B(max) = 84.5 +/- 1.6 fmol/mg of protein). The rank order of binding competition potency (K(i)) for prototypical nicotinic compounds was alpha-conotoxin MII (6 nM) > nicotine (156 nM) approximately methyllycaconitine (200 nM) > alpha-bungarotoxin (>10 microM), similar to that for nAChR in dopamine neurons displaying a distinctive pharmacology. 6-Hydroxydopamine lesioning studies indicated that beta3 and alpha5 subunits are likely partners of the alpha6 subunits in nAChR expressed in dopaminergic cell bodies. Similar to findings in rodents, quantitative real-time reverse transcription-polymerase chain reactions of human brain indicated that alpha6 subunit mRNA expression was 13-fold higher in the substantia nigra than in the cortex or the rest of the brain. Thus, heterologous expression studies suggest that the human alpha5 subunit makes a critical contribution to alpha6beta4beta3alpha5 nAChR assembly into a ligand-binding form with native alpha6(*)-nAChR-like pharmacology and of potential physiological and pathophysiological relevance. PMID- 15356218 TI - Effect of dextrometorphan and dextrorphan on nicotine and neuronal nicotinic receptors: in vitro and in vivo selectivity. AB - The effects of dextrometorphan and its metabolite dextrorphan on nicotine-induced antinociception in two acute thermal pain assays after systematic administration were evaluated in mice and compared with that of mecamylamine. Dextrometorphan and dextrorphan were found to block nicotine's antinociception in the tail-flick and hot-plate tests with different potencies (dextrometorphan is 10 times more potent than its metabolite). This blockade was not due to antagonism of N-methyl d-aspartate receptors and/or interaction with opiate receptors, since selective drugs of these receptors failed to block nicotine's analgesic effects. Our results with the tail-flick and hot-plate tests showed an interesting in vivo functional selectivity for dextrometorphan over dextrorphan. In oocytes expressing various neuronal acetylcholine nicotinic receptors (nAChR), dextrometorphan and dextrorphan blocked nicotine activation of expressed alpha(3)beta(4), alpha(4)beta(2), and alpha(7) subtypes with a small degree of selectivity. However, the in vivo antagonistic potency of dextrometorphan and dextrorphan in the pain tests does not correlate well with their in vitro blockade potency at expressed nAChR subtypes. Furthermore, the apparent in vivo selectivity of dextrometorphan over dextrorphan is not related to its in vitro potency and does suggest the involvement of other mechanisms. In that respect, dextrometorphan seems to behave as another mecamylamine, a noncompetitive nicotinic receptor antagonist with a preferential activity to alpha(3)beta(4)(*) neuronal nAChR subtypes. PMID- 15356219 TI - Biological effects of wood ash application to forest and aquatic ecosystems. AB - The present review aims to summarize current knowledge in the topic of wood ash application to boreal forest and aquatic ecosystems, and the different effects derived from these actions. Much research has been conducted regarding the effects of wood ash application on forest growth. Present studies show that, generally speaking, forest growth can be increased on wood ash-ameliorated peatland rich in nitrogen. On mineral soils, however, no change or even decreased growth have been reported. The effects on ground vegetation are not very clear, as well as the effects on fungi, soil microbes, and soil-decomposing animals. The discrepancies between different studies are for the most part explained by abiotic factors such as variation in fertility among sites, different degrees of stabilization, and wood ash dosage used, and different time scales among different studies. The lack of knowledge in the field of aquatic ecosystems and their response to ash application is an important issue for future research. The few studies conducted have mainly considered changes in water chemistry. The biotoxic effects of ash application can roughly be divided into two categories: primary and secondary. Among the primary effects is toxicity deriving from compounds in the wood ash and cadmium is probably the worst among these. The secondary effects of wood ash are generally due to its alkaline capacity and a release of ions into the soil and soil water, and finally, watercourses and lakes. Given current knowledge, we would recommend site- and wood ash-specific application practices, rather than broad and general guidelines for wood ash application to forests. PMID- 15356220 TI - Nutrient input and removal trends for agricultural soils in nine geographic regions in Arkansas. AB - Knowledge of the balance between nutrient inputs and removals is required for identifying regions that possess an excess or deficit of nutrients. This assessment describes the balance between the agricultural nutrient inputs and removals for nine geographical districts within Arkansas from 1997 to 2001. The total N, P, and K inputs were summed for each district and included inorganic fertilizer and collectable nutrients excreted as poultry, turkey, dairy, and hog manures. Nutrients removed by harvested crops were summed and subtracted from total nutrient inputs to calculate the net nutrient balance. The net balances for N, P, and K were distributed across the hectarage used for row crop, hay, pasture, or combinations of these land uses. Row-crop agriculture predominates in the eastern one-third and animal agriculture predominates in the western two thirds of Arkansas. Nutrients derived from poultry litter accounted for >92% of the total transportable manure N, P, and K. The three districts in the eastern one-third of Arkansas contained 95% of the row-crop hectarage and had net N and P balances that were near zero or negative. The six districts in the western two thirds of Arkansas accounted for 89 to 100% of the animal populations, had positive net balances for N and P, and excess P ranged from 1 to 9 kg P ha(-1) when distributed across row-crop, hay, and pasture hectarage. Transport of excess nutrients, primarily in poultry litter, outside of the districts in western Arkansas is needed to achieve a balance between soil inputs and removals of P and N. PMID- 15356221 TI - Environmental concentrations of agricultural herbicides in Saskatchewan, Canada: bromoxynil, dicamba, diclofop, MCPA, and trifluralin. AB - Herbicides are the most commonly used group of agricultural pesticides on the Canadian Prairies and, in 1990, more than 20000 Mg of herbicides were applied in the provinces of Alberta, Saskatchewan, and Manitoba. The present paper reports on environmental concentrations of five herbicides currently used in the prairie region. The herbicides bromoxynil [3,5-dibromo-4-hydroxy-benzonitrile], dicamba [3,6-dichloro-o-anisic acid], diclofop [(RS)-2-[4-(2,4-dichlorophenoxy) phenoxy]propanoic acid], MCPA [(4-chloro-2-methylphenoxy)acetic acid], and trifluralin [alpha,alpha,alpha-trifluoro-2,6-dinitro-N,N-isopropyl-p-toluidine] were measured in the atmosphere, bulk atmospheric deposits, surface film, and dugout (pond) water at two sites near Regina, Saskatchewan, during 1989 and 1990. All five herbicides were detected in air and surface film and all but trifluralin were detected in the bulk atmospheric deposits and dugout water. Trifluralin was most frequently detected in air (79% of samples) whereas bromoxynil was present in maximum concentration (4.2 ng m(-3)). MCPA was present in maximum levels in bulk atmospheric (wet plus dry) deposits (2350 ng m(-2) d(-1)), surface film (390 ng m(-2)), and dugout water (330 ng L(-1)), whereas dicamba was most frequently detected in surface film (47%) and dugout water (97%). The highest quantities of the herbicides tended to be present during or immediately after the time of regional application. PMID- 15356222 TI - An improved description of pesticide volatilization: refinement of the pesticide leaching model (PELMO). AB - The consideration of pesticide volatilization from soil surfaces as an integral component of pesticide fate models is of importance, especially as part of the Predicted Environmental Concentrations (PEC) models used in the registration procedures for pesticides. The Pesticide Leaching Model (PELMO), which is used in the European registration process, was modified to allow for a reliable prediction of volatilization from soil. The previous PELMO version was upgraded by improving the spatiotemporal discretization at the soil surface, improving the empirical description of temperature dependence of Henry's law constants and including increased sorption of pesticides in dry soils. Comparison of predictions with experimental findings revealed the improvements of PELMO to contribute to a more realistic reflection of measurements, particularly at initial stages of the studies. The broad range of literature values of Henry's law constants was shown to have a significant effect on predicted volatilization fluxes. As a main refinement, the tendency of pesticides toward enhanced volatilization under moist conditions was correctly calculated by the improved model. Variations between model predictions and measurements were due to a lack of experimental data on soil sorption under dry conditions and indicated the need for further calibration of the model. The description of water content in the top layer was subject to uncertainty, which was exemplified by an overestimation of soil moisture during the last days of the field study. Thus, future model improvement will be dependent on experimental support to obtain more detailed information on soil-air-water partitioning of pesticides in the top soil layer. PMID- 15356223 TI - Accumulation of perchlorate in aquatic and terrestrial plants at a field scale. AB - Previous laboratory-scale studies have documented perchlorate ClO(-)(4) uptake by different plant species, but less information is available at field scale, where ClO(-)(4) uptake may be affected by environmental conditions, such as distance to streams or shallow water tables, exposure duration, and species. This study examined uptake of ClO(-)(4) in smartweed (Polygonum spp.) and watercress (Nasturtium spp.) as well as more than forty trees, including ash (Fraxinus greggii A. Gray), chinaberry (Melia azedarach L.), elm (Ulmus parvifolia Jacq.), willow (Salix nigra Marshall), mulberry [Broussonetia papyrifera (L.) Vent.], and hackberry (Celtis laevigata Willd.) from multiple streams surrounding a perchlorate-contaminated site. Results indicate a large potential for ClO(-)(4) accumulation in aquatic and terrestrial plants, with ClO(-)(4) concentration in plant tissues approximately 100 times higher than that in bulk water. Perchlorate accumulation in leaves of terrestrial plants was also dependent on species, with hackberry, willow, and elm having a strong potential to accumulate ClO(-)(4). Generally, trees located closer to the stream had a higher ClO(-)(4) accumulation than trees located farther away from the stream. Seasonal leaf sampling of terrestrial plants indicated that ClO(-)(4) accumulation also was affected by exposure duration, with highest accumulation observed in the late growing cycle, although leaf concentrations for a given tree were highly variable. Perchlorate may be re-released into the environment via leaching and rainfall as indicated by lower perchlorate concentrations in collected leaf litter. Information obtained from this study will be helpful to understand the fate of ClO(-)(4) in macrophytes and natural systems. PMID- 15356224 TI - Quantification of the effect of fumigation on short- and long-term nitrogen mineralization and nitrification in different soils. AB - The effect of soil fumigation on N mineralization and nitrification needs to be better quantified to optimize N fertilizer advice and predict NO(-)(3) concentrations in crops and NO(-)(3) leaching risks. Seven soils representing a range in soil texture and organic matter contents were fumigated with Cyanamid DD 95 (a mixture of 1,3-dichloropropane and 1,3-dichloropropene). After removal of the fumigant, the fumigated soils and unfumigated controls were incubated for 20 wk and N mineralization and nitrification were monitored by destructive sampling. The average short-term N mineralization rates (k(s)) were significantly larger in the fumigated than in the unfumigated soils (P = 0.025), but the differences in k(s) between fumigated and unfumigated soils could not be related to soil properties. The average long-term N mineralization rates (k(l)) were slightly larger in the fumigated soils but the difference with the unfumigated soils was not significant. Again, the differences in k(l) values could not be related to soil properties. Nitrification was inhibited completely for at least 3 wk in all soils, and an effect on nitrification could be observed up to 17 wk in one soil. An S-shaped function was fitted to the nitrification data corrected for N mineralization, and both the rate constant (gamma) and the time at which maximum nitrification was reached (t(max)) were strongly correlated to soil pH. However, since no correlations were found between the effect of fumigation on N mineralization and soil properties, taking into account the effects of fumigation in fertilizer advice and in the prediction of NO(-)(3) leaching risks will need further research. PMID- 15356225 TI - Enzyme activities and arylsulfatase protein content of dust and the soil source: biochemical fingerprints? AB - Little is known about the potential of enzyme activities, which are sensitive to soil properties and management, for the characterization of dust properties. Enzyme activities may be among the dust properties key to identifying the soil source of dust. We generated dust (27 and 7 microm) under controlled laboratory conditions from agricultural soils (0-5 cm) with history of continuous cotton (Gossypium hirsutum L.) or cotton rotated with peanut (Arachis hypogaea L.), sorghum [Sorghum bicolor (L.) Moench], rye (Secale cereale L.), or wheat (Triticum aestivum L.) under different water management (irrigated or dryland) and tillage (conservation or conventional) systems. The 27- and 7-microm dust samples showed activities of beta-glucosidase, alkaline phosphatase, and arylsulfatase, which are related to cellulose degradation and phosphorus and sulfur mineralization in soil, respectively. Dust samples generated from a loam and sandy clay loam showed higher enzyme activities compared with dust samples from a fine sandy loam. Enzyme activities of dust samples were significantly correlated to the activities of the soil source with r > 0.74 (P < 0.01). The arylsulfatase proteins contents of the soils (0.04-0.65 mg protein kg(-1) soil) were lower than values reported for soils from other regions, but still dust contained arylsulfatase protein. The three enzyme activities studied, as a group, separated the dust samples due to the crop rotation or tillage practice history of the soil source. The results indicated that the enzyme activities of dust will aid in providing better characterization of dust properties and expanding our understanding of soil and air quality impacts related to wind erosion. PMID- 15356226 TI - Changes in soil nitrogen and phosphorus under different broiler production systems. AB - In a field study, soils of four conventional free-range and organic broiler runs were analyzed for N and P concentrations in the years 2000 and 2001. Zones of different use intensity by broilers were identified on the free runs and mean zonal nutrient contents were compared with each other. Intensity of use by birds and spatial distribution of soil nutrient concentrations were found to be related to each other. Fecal N input by broilers resulted in accumulation of soil mineral nitrogen (N(min)) contents down to a 90-cm sampling depth. In highly frequented "hot spots," plant requirement as defined by the German "N-Basis-Sollwert" (110 kg/ha N(min)) for grassland was exceeded in all four cases. This implies an increased environmental risk of ammonia volatilization and nitrate leaching. Fecal P input by broilers resulted in accumulation of plant-available and thus mobile soil P (phosphorus extracted with calcium-acetate-lactate [P(CAL)] and phosphorus extracted with water [P(w)]) in the most intensely used zones. In these areas, soil P contents exceeded 90 mg/kg P(CAL) (upper limit of soil test P defined in Germany for optimum plant yield) by as much as 217 mg/kg, which indicates an enhanced risk of P loss from the soil via runoff or leaching. The conclusion might be drawn that, with regard to nutrient loss from free-run soils, intensive indoor production in a closed system may be more environmentally neutral than conventional free-range or organic production. However, to put this into perspective, the scope of the environmental risk connected with spatially limited point accumulation of nutrients should be considered. Furthermore, an environmental evaluation must also account for the fate and environmental effects of the broiler litter produced inside the broiler house. PMID- 15356227 TI - Regional analysis of climate, primary production, and livestock density in inner Mongolia. AB - Overstocking is believed to be one of the principal causes for grassland degradation in northern China. For this reason, quantification of overstocking and spatiotemporal analysis are needed for this area. In this study, the relationship between annual rainfall and grassland aboveground net primary production (ANPP) was analyzed using data from 1982 to 1991 in the Inner Mongolia Autonomous Region (IMAR), China. Subsequently, rainfall-based livestock carrying capacity was estimated and combined with livestock density calculated from county level livestock data from 1982 to 1991 to determine spatial and temporal patterns of a stocking rate index and its relationship with climatic factors. The results indicate the following. First, there was a significant linear relationship between annual rainfall and ANPP in IMAR and the slope of ANPP versus rainfall was greater than those found in South America and Africa, indicating higher rain use efficiency. Second, temporally averaged livestock density showed overstocking in most of the rural counties except for those in the cold north, where human populations are low and transportation systems are poor. Third, the stocking rate index increased with temperature, from less than 1.0 in the north, to greater than 2.0 in most of the southern IMAR. Within the central IMAR, the index increased from west to east, along the gradient of increasing rainfall. Fourth, long-term dynamics of livestock density depicted continuous overstocking, more than 20%, from 1982 to 1991 along the western part of the NorthEast China Transect (NECT) within IMAR. Spatial planning of livestock densities according to carrying capacities and improved pastoral management are needed in this area. PMID- 15356228 TI - Sorption and degradation of alachlor and metolachlor in ground water using green sands. AB - Reactive barriers are used for in situ treatment of contaminated ground water. Waste green sand, a by-product of gray-iron foundries that contains iron particles and organic carbon, was evaluated in this study as a low-cost reactive material for treating ground water contaminated with the herbicides alachlor [2 chloro-2',6'-diethyl-N-(methoxymethyl)acetanilide] and metolachlor [2-chloro-6' ethyl-N-(2-methoxy-1-methylethyl)-o-acetoluidide]. Batch and column tests were conducted with 11 green sands to determine transport parameters and reaction rate constants for the herbicides. Similar Fe-normalized rate constants (K(SA)) were obtained from the batch and column tests. The K(SA) values obtained for green sand iron were also found to be comparable with or slightly higher than K(SA) values for Peerless iron, a common reactive medium used in reactive barriers. Partition coefficients ranging between 3.6 and 50.2 L/kg were obtained for alachlor and between 1.0 and 54.8 L/kg for metolachlor, indicating that the organic carbon and clay in green sands can significantly retard the movement of the herbicides. Partition coefficients obtained from the batch and column tests were similar (+/-25%), but the batch tests typically yielded higher partition coefficients for green sands exhibiting greater sorption. Calculations made using transport parameters from the column tests indicate that a 1-m-thick reactive barrier will result in a 10-fold reduction in concentration of alachlor and metolachlor for seepage velocities less than 0.1 m/d provided the green sand contains at least 2% iron. This level of reduction generally is sufficient to reduce alachlor and metolachlor concentrations below maximum contaminant levels in the United States. PMID- 15356229 TI - Forecasting the number of soil samples required to reduce remediation cost uncertainty. AB - Sampling scheme design is an important step in the management of polluted sites. It largely controls the accuracy of remediation cost estimates. In practice, however, sampling is seldom designed to comply with a given level of remediation cost uncertainty. In this paper, we present a new technique that allows one to estimate of the number of samples that should be taken at a given stage of investigation to reach a forecasted level of accuracy. The uncertainty is expressed both in terms of volume of polluted soil and overall cost of remediation. This technique provides a flexible tool for decision makers to define the amount of investigation worth conducting from an environmental and financial perspective. The technique is based on nonlinear geostatistics (conditional simulations) to estimate the volume of soil that requires remediation and excavation and on a function allowing estimation of the total cost of remediation (including investigations). The geostatistical estimation accounts for support effect, information effect, and sampling errors. The cost calculation includes mainly investigation, excavation, remediation, and transportation. The application of the technique on a former smelting work site (lead pollution) demonstrates how the tool can be used. In this example, the forecasted volumetric uncertainty decreases rapidly for a relatively small number of samples (20-50) and then reaches a plateau (after 100 samples). The uncertainty related to the total remediation cost decreases while the expected total cost increases. Based on these forecasts, we show how a risk-prone decision maker would probably decide to take 50 additional samples while a risk-averse decision maker would take 100 samples. PMID- 15356230 TI - Kinetics of chromate adsorption on goethite in the presence of sorbed silicic acid. AB - The adsorption of chromate on mineral surfaces has received much attention due to its toxicity in natural systems. Spectroscopic studies have demonstrated that chromate forms inner-sphere complexes on variable-charge surfaces. However, in natural systems chromate has been observed to be fairly mobile, which has been explained by the presence of naturally occurring ligands competing with chromate for mineral surface sites. Silicic acid is a ubiquitous ligand in soil and water environments and also sorbs strongly to variable-charge surfaces. Yet little research has examined its influence on chromate adsorption to variable-charge surfaces such as goethite. This study examined the influence of silicic acid (0.10 and 1.0 mM) on the adsorption kinetics of chromate (0.05 and 0.10 mM) on goethite over a range of common soil pH values (4, 6, and 8). The rate and total quantity of chromate adsorption decreased in all the experiments except at a pH value of 4 and a chromate concentration of 0.05 mM. The inhibition of chromate adsorption ranged from 3.1% (pH = 4, Si = 0.10 mM, chromate = 0.10 mM) to 83.3% (pH = 8, Si = 1.0 mM, chromate = 0.05 mM). The rate of chromate adsorption decreased with an increase in pH and silicic acid concentration. This was attributed to a reduction in the surface potential of goethite on silicic acid adsorption as well as a competition for surface sites. The presence of naturally occurring ligands such as silicic acid may be responsible for the enhanced mobility of chromate in natural systems and demonstrates the importance of competitive adsorption for evaluating the mobility of trace elements. PMID- 15356231 TI - Phosphorus loss and runoff characteristics in three adjacent agricultural watersheds with claypan soils. AB - Effects of precipitation, runoff, and management on total phosphorus (TP) loss from three adjacent, row-cropped watersheds in the claypan region of northeastern Missouri were examined from 1991 to 1997 to understand factors affecting P loss in watersheds dominated by claypan soils. Runoff samples from each individual runoff event were analyzed for TP and sediment concentration. The annual TP loss ranged from 0.29 to 3.59 kg ha(-1) with a mean of 1.36 kg ha(-1) across all the watersheds during the study period. Significantly higher loss of TP from the watersheds was observed during the fallow period. Multiple small runoff events or several large runoff events contributed to loss of TP from the watersheds. Total P loss in 1993, a year with above-normal precipitation, accounted for 30% of the total TP loss observed over seven years. The five largest runoff events out of a total of 66 events observed over seven years accounted for 27% of the TP loss. The five largest sediment losses were responsible for 24% of the TP loss over seven years. Runoff volume and sediment loss explained 64 to 73% and 47 to 58% of the variation in TP loss on watersheds during the study. Flow duration and maximum flow accounted for 49 and 66% of TP loss, respectively. The results of this study suggest that management practices that reduce runoff volume, flow duration, maximum flow, and sediment loss, and that maintain a suitable vegetative cover throughout the year could lower P loss in claypan soils. PMID- 15356232 TI - Modeling the effects of tillage management practices on herbicide runoff in northern Italy. AB - The need to quantitatively predict pesticide runoff and erosion under cropping system management has gained increasing importance. In Europe, predictive models have not yet been fully validated because of the lack of field data sets. The objective of this study was to validate the capability of PRZM (Pesticide Root Zone Model) 3.12 to predict water runoff, sediment erosion, and associated transport of atrazine (6-chloro-N(2)-ethyl-N(4)-isopropyl-1,3,5-triazine-2,4 diamine), terbuthylazine (N(2)-tert-butyl-6-chloro-N(4)-ethyl-1,3,5-triazine-2,4 diamine), and metolachlor [2-chloro-6'-ethyl-N-(2-methoxy-l-methylethyl)acet-o toluidide] under common tillage management practices found in northern Italy. A 2 yr field data set was used to evaluate the model. Results showed that the model could qualitatively simulate significant differences of water runoff, soil erosion, and associated herbicide losses between conventional tillage (CT) and minimum tillage (MT) for a winter barley (Hordeum vulgare L.) cover crop. For MT, water runoff, soil erosion, herbicide losses in water runoff and eroded sediment, and the proportion of herbicide loss via sediment erosion were significantly lower than for CT. The model failed to correctly simulate event-based herbicide concentration, water runoff, and soil erosion. The model usually underestimated pesticide runoff events with high rainfall intensity and low daily precipitation volume, and overestimated runoff events with low intensity and high volume. The main reason was that the description of runoff and erosion processes is rather empirical in the model and not physically based. Moreover, model calculations do not adequately reflect the relationships between soil erosion intensity and chemical concentration in sediment losses, leading to discrepancies between predictions and field observations. PMID- 15356233 TI - Effects of mineral surfaces on pyrene partitioning to well-characterized humic substances. AB - Mineral surfaces can alter the ability of humic substances (HS) to bind hydrophobic organic contaminants. In this study, complete adsorption (i.e., to avoid HS adsorptive fractionation effects) of a small subset of well characterized terrestrial and aquatic HS on kaolinite and hematite significantly changed their subsequent organic carbon-normalized partition coefficients K(ads)(oc) for pyrene relative to their original respective dissolved organic carbon-normalized partition coefficients K(dis)(oc). Parallel experiments with ultrafiltration (UF) fractions obtained from purified Aldrich humic acid (PAHA) (Aldrich Chemical, Milwaukee, WI) gave similar results. The heterogeneity among the PAHA UF fractions was examined via their mineral surface adsorption characteristics and their subsequent ability to bind pyrene. As expected, variations in maximum adsorption densities (q(max)), Langmuir adsorption constants (K(q)), and pyrene K(ads)(oc) values were observed among the PAHA UF fractions. However, general trends of q(max), K(q), and pyrene log K(ads)(oc) values for the PAHA UF fractions versus the logarithm of their weight-average molecular weights (MW(w)) did not typically match the corresponding trends obtained with the four aquatic and terrestrial HS. In general, an ideal mixture competitive adsorption model gave reasonable predictions for PAHA sorption to kaolinite and hematite based on their corresponding UF isotherm parameters. Ideal mixture predictions of pyrene partitioning to adsorbed PAHA from the corresponding UF fraction results were better for kaolinite versus hematite, indicating that the underlying mineral surface can alter the effects of HS heterogeneity on hydrophobic organic contaminant sorption. PMID- 15356234 TI - Environmental fate of triasulfuron in soils amended with municipal waste compost. AB - The amendment of soil with compost may significantly influence the mobility and persistence of pesticides and thus affect their environmental fate. Factors like adsorption, kinetics, and rate of degradation of pesticides could be altered in amended soils. The aim of this study was to determine the effects of the addition of compost made from source-separated municipal waste and green waste, on the fate of triasulfuron [(2-(2-chloroethoxy)-N-[[4-methoxy-6-methyl-1,3,5-triazin-2 yl)amino]carbonyl]benzenesulfonamide], a sulfonylurea herbicide used in postemergence treatment of cereals. Two native soils with low organic matter content were used. A series of analyses was performed to evaluate the adsorption and degradation of the herbicide in soil and in solution after the addition of compost and compost-extracted organic fractions, namely humic acids (HA), fulvic acids (FA), and hydrophobic dissolved organic matter (HoDOM). Results have shown that the adsorption of triasulfuron to soil increases in the presence of compost, and that the HA and HoDOM fractions are mainly responsible for this increase. Hydrophobic dissolved organic matter applied to the soils underwent sorption reactions with the soils, and in the sorbed state, served to increase the adsorption capacity of the soil for triasulfuron. The rate of hydrolysis of triasulfuron in solution was significantly higher at acidic pH and the presence of organic matter fractions extracted from compost also slightly increased the rate of hydrolysis. The rate of degradation in amended and nonamended soils is explained by a two-stage degradation kinetics. During the initial phase, although triasulfuron degradation was rapid with a half-life of approximately 30 d, the presence of compost and HoDOM was found to slightly reduce the rate of degradation with respect to that in nonamended soil. PMID- 15356235 TI - Distribution of polychlorinated biphenyls and chlorinated pesticide residues in trout in the Sierra Nevada. AB - Organochlorine compounds are known to be atmospherically transported to long distances from their original sources. To understand the influence of California's Sierra Nevada range on the air transport and subsequent distribution pattern of some of these residues within the range, we have chosen salmonid fish as an indicator species. Fish were collected from 10 locations throughout the northern and central Sierra Nevada and polychlorinated biphenyl (PCB), toxaphene, chlordane, and DDT [1,1,1-trichloro, 2,2'-bis (p-chlorophenyl) ethane] residues in muscle tissue were analyzed. Rainbow trout (Oncorhynchus mykiss) were found in all sampling locations, and therefore analyses mainly focused on this species. When similar-sized rainbow trout samples from several similar oligotrophic, high altitude lakes and streams were compared, it became apparent that altitude is one of the factors affecting the residual levels of PCB (r(2) = 0.882), but not for total DDT, toxaphene, or chlordane in trout. Analysis of correlations among these four organochlorine compound residue groups indicated that there are modest correlations in patterns of distribution between chlordane vs. toxaphene (r(2) = 0.345), and chlordane vs. total DDT (r(2) = 0.239), but toxaphene residues are not correlated with PCB or total DDT. In view of significant correlation to the altitude it is concluded that PCB residue in rainbow trout is a good monitoring tool for studying the effect of high-altitude mountain ranges on the long-range transport and distribution of those persistent pollutants. PMID- 15356236 TI - Interactions of chlorpyrifos with colloidal materials in aqueous systems. AB - An understanding of sorptive processes is key to describing the fate of chlorpyrifos [O,O-diethyl-O-(3, 5, 6-trichloro-2-pyridyl) phosphorothioate] in aquatic environments. The objectives of this study were to evaluate isotherms for adsorption and desorption of chlorpyrifos on colloidal materials and to advance understanding of interaction mechanisms between chlorpyrifos and colloidal materials. Six Ca-saturated reference smectites, one Ca-saturated humic acid (Ca humate), and one suspended sediment sample, collected from the Upper Cedar River, Iowa, were studied. A batch equilibration technique was employed to quantify adsorption and desorption isotherms for chlorpyrifos over the 0 to 100 microg L( 1) concentration range in a 0.01 M CaCl(2) background. Large differences in sorption affinity and variation in desorption hysteresis were found among the smectites. Neither chlorpyrifos adsorption nor its desorption were correlated with cation exchange capacity, surface area, or surface charge density of the smectites. The evidence suggests that physical interaction between chlorpyrifos and smectites is the dominant mechanism for adsorption of chlorpyrifos in aqueous systems. Chlorpyrifos was very strongly sorbed on Ca-humate and was not desorbed from the Ca-humate back into the aqueous solution. Chlorpyrifos was moderately sorbed on river sediment, and a large adsorption-desorption hysteresis was also found. The study implies that the nature of both organic and inorganic materials in suspended sediment can influence the adsorption-desorption behavior of chlorpyrifos in aqueous systems. PMID- 15356237 TI - Degradation of N,N'-dibutylurea (DBU) in soils treated with only DBU and DBU fortified benlate fungicides. AB - N,N'-dibutylurea (DBU) is a breakdown product of benomyl [methyl 1 (butylcarbamoyl)-2-benzimidazole carbamate], the active ingredient in Benlate fungicides, and has been proposed to cause crop damage after the use of Benlate 50 DF fungicide (DuPont, Wilmington, DE). Our research focused on DBU persistence after application into soil. We assessed DBU persistence on direct application of DBU (carbonyl-(14)C) at two concentrations (0.08 and 0.8 microg DBU kg(-1)) to seven soils and two potting mixes in soil microcosms incubated at various combinations of soil water potential (-0.03 or -0.1 MPa) and temperature (23, 33, 44 degrees C). For two soils at a subset of treatment variables we assessed DBU persistence in the presence of Benlate DF and SP fungicide formulations. Parent compounds, metabolites, and (14)CO(2) were tracked using chromatographic analysis with radioassay and UV detection, liquid scintillation counting, and post extraction oxidation of the soil. DBU degradation was primarily microbial and for most soil-treatment combinations, half-lives were less than 2 wk. DBU degradation was retarded at the lower soil water potential and enhanced at 33 degrees C. In the presence of the formulation, DBU degradation was slower for one soil type. The longest half-life observed in any case was less than 7 wk; therefore, long term persistence of DBU applied to soils through a Benlate application is very unlikely. PMID- 15356238 TI - Clonal differences in mercury tolerance, accumulation, and distribution in willow. AB - This study was performed to investigate mercury (Hg) tolerance, accumulation, and translocation within the genus Salix for the potential use of this plant to remediate Hg-contaminated sites. Six clones of willow (Salix spp.) were tested on tolerance to Hg by treating plants grown in solution culture with 0 to 15 microM HgCl(2). Results showed that willow had a large variation in its sensitivity to Hg. However, the accumulation and translocation of Hg to shoots was similar in the eight tested willow clones as shown by cold vapor atomic absorption spectrometry analysis when plants were treated with 0.5 microM HgCl(2) in a nutrient solution. The majority of total Hg accumulated was localized to the roots, whereas only 0.45 to 0.62% of the total Hg accumulated via roots was translocated to the shoots. Thus, the root system is the main tissue of willow that accumulates Hg and the majority of the Hg in the root system (80%) was bound in the cell wall. PMID- 15356239 TI - Leaching and crop uptake of nitrogen from nitrogen-15-labeled green manures and ammonium nitrate. AB - Green manures can be used as an N source for agricultural crops as a substitute for inorganic N fertilizers. The effects of using green manures on leaching and uptake of N by spring barley (Hordeum vulgare L.) were evaluated in a 2-yr lysimeter study. Ryegrass (Lolium perenne L.) and red clover (Trifolium pratense L.) labeled with (15)N were applied in May of the first year at 160 kg total N ha(-1). Simultaneously, (15)NH(4)(15)NO(3) was applied at 80 kg N ha(-1) to additional lysimeters and others were left without N additions (control). During the second year, all lysimeters, except the control, received 80 kg N ha(-1) as unlabeled NH(4)NO(3). The cumulative, average loads of total N leached during the two years were: 37 (control), 62 (NH(4)NO(3)), 50 (ryegrass manure), and 73 (red clover manure) kg ha(-1). The differences among the treatments were not significant (P > 0.05), but the control had significantly smaller (P < 0.05) leaching loads than the treatments. About 24% of ryegrass- and red clover-derived N and 43% of NH(4)NO(3) were removed through spring barley grain and stover during the two growing seasons. Thus, the N use efficiency in barley was substantially larger when grown with inorganic N fertilizer than when grown with green manure. Viewed in combination with the tendency for larger N leaching loads under red clover manure, claims about water quality benefits of legume-based green manures should be evaluated with regard to the timing of N release and demand for N by the plant. PMID- 15356240 TI - Dissolution of phosphate in a phosphorus-enriched ultisol as affected by microbial reduction. AB - Phosphorus dissolution often increases as soils become more reduced, but the mechanisms are not fully understood. The objectives of this research were to determine rates and mechanisms of P dissolution during microbial reduction of a surface soil from the North Carolina Coastal Plain. Duplicate suspensions of silt + clay fractions from a Cape Fear sandy clay loam (fine, mixed, semiactive, thermic Typic Umbraquult) were reduced in a continuously stirred redox reactor for 40 d. We studied the effects of three treatments on P dissolution: (i) 2 g dextrose kg(-1) solids added as a microbial carbon source at time 0 d; (ii) 2 g dextrose kg(-1) solids split into three additions at 0, 12, and 26 d; and (iii) no added dextrose. After 40 d of reduction, concentrations of dissolved reactive phosphorus (DRP) were similar for all treatments and increased up to sevenfold from 1.5 to 10 mg L(-1). The initial rate of reduction and dissolution of DRP was significantly greater for the 0-d treatment. A linear relationship (R(2) = 0.79) was found between DRP and dissolved organic carbon (DOC). Dissolved Fe and Al and pH increased, suggesting the formation of aqueous Fe- and Al-organic matter complexes. Separate batch experiments were performed to study the effects of increasing pH and citrate additions on PO(4) dissolution under aerobic conditions. Increasing additions of citrate increased concentrations of DRP, Fe, and Al, while increasing pH had no effect. Results indicated that increased dissolved organic matter (DOM) during soil reduction contributed to the increase in DRP, perhaps by competitive adsorption or formation of aqueous ternary DOM-Fe PO(4) or DOM-Al-PO(4) complexes. PMID- 15356241 TI - Nitrate leaching to subsurface drains as affected by drain spacing and changes in crop production system. AB - Subsurface drainage is a beneficial water management practice in poorly drained soils but may also contribute substantial nitrate N loads to surface waters. This paper summarizes results from a 15-yr drainage study in Indiana that includes three drain spacings (5, 10, and 20 m) managed for 10 yr with chisel tillage in monoculture corn (Zea mays L.) and currently managed under a no-till corn-soybean [Glycine max (L.) Merr.] rotation. In general, drainflow and nitrate N losses per unit area were greater for narrower drain spacings. Drainflow removed between 8 and 26% of annual rainfall, depending on year and drain spacing. Nitrate N concentrations in drainflow did not vary with spacing, but concentrations have significantly decreased from the beginning to the end of the experiment. Flow weighted mean concentrations decreased from 28 mg L(-1) in the 1986-1988 period to 8 mg L(-1) in the 1997-1999 period. The reduction in concentration was due to both a reduction in fertilizer N rates over the study period and to the addition of a winter cover crop as a "trap crop" after corn in the corn-soybean rotation. Annual nitrate N loads decreased from 38 kg ha(-1) in the 1986-1988 period to 15 kg ha(-1) in the 1997-1999 period. Most of the nitrate N losses occurred during the fallow season, when most of the drainage occurred. Results of this study underscore the necessity of long-term research on different soil types and in different climatic zones, to develop appropriate management strategies for both economic crop production and protection of environmental quality. PMID- 15356242 TI - Phosphorus in surface runoff from calcareous arable soils of the semiarid Western United States. AB - Management strategies that minimize P transfer from agricultural land to water bodies are based on relationships between P concentrations in soil and runoff. This study evaluated such relationships for surface runoff generated by simulated sprinkler irrigation onto calcareous arable soils of the semiarid western United States. Irrigation was applied at 70 mm h(-1) to plots on four soils containing a wide range of extractable P concentrations. Two irrigation events were conducted on each plot, first onto dry soil and then after 24 h onto wet soil. Particulate P (>0.45 microm) was the dominant fraction in surface runoff from all soils and was strongly correlated with suspended sediment concentration. For individual soil types, filterable reactive P (<0.45 microm) concentrations were strongly correlated with all soil-test P methods, including environmental tests involving extraction with water (1:10 and 1:200 soil to solution ratio), 0.01 M CaCl(2), and iron strips. However, only the Olsen-P agronomic soil-test procedure gave models that were not significantly different among soils. Soil chemical differences, including lower CaCO(3) and water-extractable Ca, higher water extractable Fe, and higher pH, appeared to account for differences in filterable reactive P concentrations in runoff from soils with similar extractable P concentrations. It may therefore be possible to use a single agronomic test to predict filterable reactive P concentrations in surface runoff from calcareous soils, but inherent dangers exist in assuming a consistent response, even for one soil within a single field. PMID- 15356243 TI - Nitrogen fertilizer form and associated nitrate leaching from cool-season lawn turf. AB - Various N fertilizer sources are available for lawn turf. Few field studies, however, have determined the losses of nitrate (NO(3)-N) from lawns receiving different formulations of N fertilizers. The objectives of this study were to determine the differences in NO(3)-N leaching losses among various N fertilizer sources and to ascertain when losses were most likely to occur. The field experiment was set out in a completely random design on a turf typical of the lawns in southern New England. Treatments consisted of four fertilizer sources with fast- and slow-release N formulations: (i) ammonium nitrate (AN), (ii) polymer-coated sulfur-coated urea (PCSCU), (iii) organic product, and (iv) a nonfertilized control. The experiment was conducted across three years and fertilized to supply a total of 147 kg N ha(-1) yr(-1). Percolate was collected with zero-tension lysimeters. Flow-weighted NO(3)-N concentrations were 4.6, 0.57, 0.31, and 0.18 mg L(-1) for AN, PCSCU, organic, and the control, respectively. After correcting for control losses, average annual NO(3)-N leaching losses as a percentage of N applied were 16.8% for AN, 1.7% for PCSCU, and 0.6% for organic. Results indicate that NO(3)-N leaching losses from lawn turf in southern New England occur primarily during the late fall through the early spring. To reduce the threat of NO(3)-N leaching losses, lawn turf fertilizers should be formulated with a larger percentage of slow-release N than soluble N. PMID- 15356244 TI - Effects of aeration on water quality from septic system leachfields. AB - We conducted a pilot-scale study at a research facility in southeastern Connecticut to assess the effects of leachfield aeration on removal of nutrients and pathogens from septic system effluent. Treatments consisted of lysimeters periodically aerated to maintain a headspace O(2) concentration of 0.209 mol mol( 1) (AIR) or vented to an adjacent leachfield trench (LEACH) and were replicated three times. All lysimeters were dosed with effluent from a septic tank for 24 mo at a rate of 12 cm d(-1) and subsequently for 2 mo at 4 cm d(-1). LEACH lysimeters had developed a clogging mat, or biomat, 20 mo before the beginning of our study. The level of aeration in the AIR treatment was held constant regardless of loading rate. No conventional biomat developed in the AIR treatment, whereas a biomat was present in the LEACH lysimeters. The headspace of LEACH lysimeters was considerably depleted in O(2) and enriched in CH(4), CO(2), and H(2)S relative to AIR lysimeters. Drainage water from AIR lysimeters was saturated with O(2) and had significantly lower pH, five-day biological oxygen demand (BOD(5)), and ammonium, and higher levels of nitrate and sulfate than LEACH lysimeters regardless of dosing rate. By contrast, significantly lower levels of total N and fecal coliform bacteria were observed in AIR than in LEACH lysimeters only at the higher dosing rate. No significant differences in total P removal were observed. Our results suggest that aeration may improve the removal of nitrogen, BOD(5), and fecal coliforms in leachfield soil, even in the absence of a biomat. PMID- 15356245 TI - Rate of fall-applied liquid swine manure: effects on runoff transport of sediment and phosphorus. AB - Reducing the delivery of phosphorus (P) from land-applied manure to surface water is a priority in many watersheds. Manure application rate can be controlled to manage the risk of water quality degradation. The objective of this study was to evaluate how application rate of liquid swine manure affects the transport of sediment and P in runoff. Liquid swine manure was land-applied and incorporated annually in the fall to runoff plots near Morris, Minnesota. Manure application rates were 0, 0.5, 1, and 2 times the rate recommended to supply P for a corn (Zea mays L.)-soybean [Glycine max (L.) Merr.] rotation. Runoff volume, sediment, and P transport from snowmelt and rainfall were monitored for 3 yr. When manure was applied at the highest rate, runoff volume and sediment loss were less than the control plots without manure. Reductions in runoff volume and soil loss were not observed for spring runoff when frozen soil conditions controlled infiltration rates. The reduced runoff and sediment loss from manure amended soils compensated for addition of P, resulting in similar runoff losses of total P among manure application rates. However, losses of dissolved P increased with increasing manure application rate for runoff during the spring thaw period. Evaluation of water quality risks from fall-applied manure should contrast the potential P losses in snowmelt runoff with the potential that incorporated manure may reduce runoff and soil loss during the summer. PMID- 15356246 TI - Effect of poultry diet on phosphorus in runoff from soils amended with poultry manure and compost. AB - Phosphorus in runoff from fields where poultry litter is surface-applied is an environmental concern. We investigated the effect of adding phytase and reducing supplemental P in poultry diets and composting poultry manures, with and without Fe and Al amendments, on P in manures, composts, and runoff. We used four diets: normal (no phytase) with 0.4% supplemental P, normal + phytase, phytase + 0.3% P, and phytase + 0.2% P. Adding phytase and decreasing supplemental P in diets reduced total P but increased water-extractable P in manure. Compared with manures, composting reduced both total P, due to dilution of manure with woodchips and straw, and water-extractable P, but beyond a dilution effect so that the ratio of water-extractable P to total P was less in compost than manure. Adding Fe and Al during composting did not consistently change total P or water extractable P. Manures and composts were surface-applied to soil boxes at a rate of 50 kg total P ha(-1) and subjected to simulated rainfall, with runoff collected for 30 min. For manures, phytase and decreased P in diets had no significant effect on total P or molybdate-reactive P loads (kg ha(-1)) in runoff. Composting reduced total P and molybdate-reactive P loads in runoff, and adding Fe and Al to compost reduced total P but not molybdate-reactive P loads in runoff. Molybdate-reactive P in runoff (mg box(-1)) was well correlated to water extractable P applied to boxes (mg box(-1)) in manures and composts. Therefore, the final environmental impact of dietary phytase will depend on the management of poultry diets, manure, and farm-scale P balances. PMID- 15356247 TI - Nitrogen, phosphorus, and potassium uptake by wheat and their distribution in soil following successive, annual compost applications. AB - The overall objective of the present study was to determine the loading limits of composts that should be applied annually to irrigated wheat. We conducted a container experiment in a greenhouse during four years. It included eight treatments: sewage sludge compost (SSC) and cattle manure compost (CMC), each applied annually to a sandy soil, at rates equivalent to 3, 6, and 12 kg m(-2), and two controls, one fertilized and one unfertilized. Total dry matter (DM), grain production, and the amount of N, P, and K taken up by plants increased with increasing compost rate. Nitrogen uptake by the plants of the fertilized control was much higher than by the plants of the highest compost rate. Phosphorus and K uptake by the plants amended with the highest compost rate was much higher than by the fertilized control plants. Inorganic N quantity in the soil increased with increasing compost rate and with successive applications. The net N mineralization during the first year of wheat growth was very low, less than 3.5% of the applied organic N under all compost application rates. The contribution of the organic N mineralization increased during the second and third years. Most of the N increase in the compost treatment was found in the upper layer of 0 to 15 cm, whereas in the fertilized treatment N accumulated from the surface to the bottom of the container, 0 to 55 cm. The successive application of high rates of composts resulted in P and K accumulation in the soil profile. PMID- 15356248 TI - Dynamic respiration index as a descriptor of the biological stability of organic wastes. AB - Analytical methods applicable to different organic wastes are needed to establish the extent to which readily biodegradable organic matter has decomposed (i.e., biological stability). The objective of this study was to test a new respirometric method for biological stability determination of organic wastes. Dynamic respiration index (DRI) measurements were performed on 16 organic wastes of different origin, composition, and biological stability degree to validate the test method and result expression, and to propose biological stability limits. In addition, theoretical DRI trends were obtained by using a mathematical model. Each test lasted 96 h in a 148-L-capacity respirometer apparatus, and DRI was monitored every hour. The biological stability was expressed as both single and cumulative DRI values. Results obtained indicated that DRI described biological stability in relation to waste typology and age well, revealing lower-stability waste characterized by a well-pronounced DRI profile (a marked peak was evident) that became practically flat for samples with higher biological stability. Fitting indices showed good model prediction compared with the experimental data, indicating that the method was able to reproduce the aerobic process, providing a reliable indication of the biological stability. The DRI can therefore be proposed as a useful method to measure the biological stability of organic wastes, and DRI values, calculated as a mean of 24 h of the highest microbial activity, of 1000 and 500 mg O(2) kg(-1) volatile solids (VS) h(-1) are proposed to indicate medium (e.g., fresh compost) and high (e.g., mature compost) biological stabilities, respectively. PMID- 15356249 TI - Seawater neutralization of alkaline bauxite residue and implications for revegetation. AB - Reaction of bauxite residue with seawater results in neutralization of alkalinity through precipitation of Mg-, Ca-, and Al-hydroxide and carbonate minerals. In batch studies, the initial pH neutralization reaction was rapid (<5 min), with further reaction continuing to reduce pH for several weeks. Reaction with seawater produced a residue pH of 8 to 8.5. Laboratory leaching column studies were undertaken to provide information on seawater neutralization of the coarse textured fraction of the waste, residue sand (RS), under conditions comparable with those that might be applied in the field. An 0.80-m-deep column of RS was neutralized by the application of the equivalent of 2-m depth of seawater. In addition to lowering the pH and Na content of the residue, seawater neutralization resulted in the addition of substantial amounts of the plant nutrients Ca, Mg, and K to the profile. Similar results were also obtained from a field-scale assessment of neutralization. However, the accumulation of precipitate, consisting of hydrotalcite, aragonite, and pyroaurite, in the drainage system may preclude the use of in situ seawater neutralization as a routine rehabilitation practice. Following seawater neutralization, RS remains too saline to support plant growth and would require fresh water leaching before revegetation. PMID- 15356250 TI - Relevance of organic matter fractions as predictors of wastewater sludge mineralization in soil. AB - Seventeen different wastewater sludges were characterized using both chemical and organic matter fractionation methods (water extraction, Van Soest method, and acid hydrolysis) and 6-mo incubation studies to assess their decomposition in soil. Simple correlation and multiple factor analysis (MFA) were then performed to establish relationships between composition and C and N mineralization of sludges. Carbon and N concentrations covered a wide range of values, but organic carbon (C(o)) to organic nitrogen (N(o)) ratios were relatively low (from 5 to 19). Carbon and N were mainly distributed in the most soluble fractions of the Van Soest method and in the water-insoluble fraction at 100 degrees C. Carbon mineralization varied from 180 to 661 g C kg(-1) organic C added during the 168-d incubation. The addition of sludges led to different inorganic N dynamics: from 3.3 to +120.0 g N kg(-1) sludge organic C mineralized after the 168-d incubation. Fractionation studies showed that the most discriminating method was acid hydrolysis. Carbon mineralization was linked with the proportion of sludge N and C present in the lignin-like fraction (r = -0.68 and -0.65, respectively). Significant relationships were established between N mineralization and N(o) to C(o) ratio (0.88 < r < 0.95) and the C(o) to N(o) ratio of sludges, the C to N ratio of the soluble fraction obtained by the Van Soest method, the water-soluble fraction at 100 degrees C, and the C and N present in the acid-hydrolyzable fraction. Finally, multiple factor analysis also enabled establishing a sludge typology using five clusters based on composition and mineralization characteristics. PMID- 15356251 TI - Water-soluble and solid-state speciation of phosphorus in stabilized sewage sludge. AB - Three chemicals, ferrous sulfate (Fe-sul), calcium oxide (CaO), and aluminum sulfate (alum), were used to stabilize phosphorus (P) in fresh, anaerobically digested sewage sludge (FSS). The chemically stabilized sludge materials and biosolids compost (BSC) were compared with the FSS with respect to water-soluble phosphorus (WSP) content in its inorganic (WSP(i)) and organic (WSP(o)) forms as well as water-soluble organic carbon (DOC). Solid-state P speciation was further probed by X-ray diffraction (XRD) and scanning electron microscopy (SEM) equipped with energy-dispersive X-ray elemental spectrometry (EDXS). Water-soluble P was effectively controlled by a wide range of Fe-sul or CaO additions to the sludge (Ca to P ratio = 3.47-17.72, Fe to P ratio = 1.01-16.53), but by only a narrow range (Al to P ratio = 1.04-2.87) of alum addition. The WSP content in the BSC was also depressed, but to a lesser extent. The pH in the treated sludge ranged from 3.0 to 12.5 and served as a key factor to control P chemistry. No correlation was observed between DOC and WSP(o). No crystallized Ca-P minerals were detected in the CaO-stabilized sludge, but brushite crystallization seemed to be obtained by low addition of Fe-sul and alum. Variscite and strengite crystallization was obtained following high addition of Fe-sul or alum, as detected by XRD and SEM-EDXS. Adsorption of P by newly formed Fe-hydroxide seems to play an important role in the Fe-sul-stabilized sludge. We concluded that administration of the tested chemicals at the proper rate can effectively reduce the hazard of P release and leaching from sludge. PMID- 15356252 TI - Alum amendment effects on phosphorus release and distribution in poultry litter amended sandy soils. AB - Increased poultry production has contributed to excess nutrient problems in Atlantic Coastal Plain soils due to land application of poultry litter (PL). Aluminum sulfate [alum, Al(2)(SO(4))(3).14H(2)O] amendment of PL effectively reduces soluble phosphorus (P) in the PL; however, the effects of these litters when added to acidic, sandy soils are not well understood. The objective of this study was to investigate the efficacy of alum-amended poultry litter in reducing P release from three Delaware Coastal Plain soils: Evesboro loamy sand (Ev; excessively drained, mesic, coated Typic Quartzipsamments), Rumford loamy sand (Ru; well drained, coarse-loamy, siliceous, subactive, thermic Typic Hapludults), and Pocomoke sandy loam (Pm; very poorly drained, coarse-loamy, siliceous, active, thermic Typic Umbraquults). Long-term (25 d) and short-term (24 h) desorption studies were conducted, in addition to chemical extractions and kinetic modeling, to observe the changes that alum-amended versus unamended PL caused in the soils. The Ev, Ru, and Pm soils were incubated with 9 Mg ha(-1) of alum-amended or unamended PL. Long-term desorption (25 d) of the incubated material resulted in approximately 13.5% (Ev), 12.7% (Ru), and 13.3% (Pm) reductions in cumulative P desorbed when comparing soil treated with unamended and alum-amended PL. In addition, the P release from the soil treated with alum amended litter was not significantly different from the control (soil alone). Short-term desorption (24 h) showed 7.3% (Ev), 15.4% (Ru), and 20% (Pm) reductions. The overall implication from this study is that the use of alum as a PL amendment is useful in coarse-textured soils of the Coastal Plain. With increased application of alum-amended PL, more significant decreases may be possible with little or no effect on soil quality. PMID- 15356253 TI - Fecal contamination of pastoral wetlands. AB - Near-channel hill-country wetlands draining steep pastoral land in New Zealand exhibit high levels of fecal contamination at a range of flows. This contamination is attributed to both the transport of bacteria into a wetland from the surrounding catchment and the direct excretion of fecal material onto wetlands by grazing cattle. E. coli concentrations observed at low to moderate flow at 20 sites varied between 0.5 x 10(1) and 2 x 10(4) most probable number (MPN) 100 mL(-1). High flow concentrations measured at two wetlands ranged up to 6 x 10(6) MPN 100 mL(-1) and yielded storm period bacterial loads of between 1 x 10(6) and 3 x 10(10) MPN per event. Given the disproportionately large fraction of surface and subsurface flow from the catchment that passes through the wetlands, these yields represent a large proportion of the total loss of bacteria from steep grazed hillsides, across a range of storm events. Cattle are attracted to the smaller, shallower wetlands for grazing in both summer and winter. Excluding stock from shallow wetlands may therefore yield improvements in bacterial water quality, although accurately quantifying this improvement is difficult without long-term studies. Cattle are not attracted to larger, deeper wetlands, presumably for fear of entrapment, and fencing them is unlikely to realize significant improvements in bacterial water quality. A statistical model incorporating solar radiation and flow explains 87% of the variance in E. coli concentrations across five monitored rainfall events. A positive correlation was found between solar radiation and E. coli concentration. The study was conducted in winter when clear, sunny days are relatively cold. Solar radiation on these days appears to be too weak to promote die-off but the colder temperatures aid survival. PMID- 15356254 TI - Comparison of three enzyme immunoassays for measuring 17beta-estradiol in flushed dairy manure wastewater. AB - Natural steroidal estrogens are an environmental concern because low nanogram per liter concentrations in water can adversely affect aquatic vertebrate species by disrupting the normal function of their endocrine systems. There is a critical need to accurately measure estrogens in dairy wastes, a potential source of estrogens such as 17beta-estradiol, to assess the risk of estrogen contamination of agricultural drainage waters resulting from land application. Commercially available enzyme immunoassay (EIA) kits have been used for measuring 17beta estradiol in livestock manure, but it is not known if different EIAs provide similar results. We compared three EIAs by measuring 17beta-estradiol in two samples of flushed dairy manure wastewater (FDMW). The measured concentrations of 17beta-estradiol in FDMW differed according to the immunoassay used. The differences were attributed to a matrix interference associated with coextracted humic substances. Future research should develop methods that enable routine measurement of 17beta-estradiol in livestock wastes by more conclusive analytical techniques such as gas chromatography-mass spectrometry. PMID- 15356255 TI - Changes in the nature of sewage sludge organic matter during a twenty-one-month incubation. AB - Six sewage sludges from five sewage treatment plants in Australia were incubated for up to 21 months. Carbon losses at the end of the 21-mo incubation varied substantially. The remaining organic matter was isolated by treatment with hydrofluoric acid (HF) and characterized using a range of solid-state (13)C nuclear magnetic resonance (NMR) spectroscopic techniques. By every measure (signal distribution in cross polarization [CP] and Bloch decay [BD] spectra, carbon NMR observability determined by spin counting, and the appearance of proton spin relaxation editing subspectra), the chemical composition of the residual organic matter appeared to be little different from that of the original sludges, even for those sludges that experienced the greatest carbon losses. Importantly, these NMR properties distinguish sewage sludge organic matter from soil organic matter. Thus, it should be possible to follow the decomposition of sewage sludge organic matter applied to soils in the field using solid-state (13)C NMR spectroscopy. PMID- 15356256 TI - Sorption and detoxification of toxic compounds by a bifunctional organoclay. AB - Organoclays are excellent sorbents for nonionic contaminants and therefore may have many environmental applications. A major limitation on the use of organoclays is that the contaminant merely changes its location from one environmental compartment to another while still remaining intact. In this study, a new type of organoclay, termed a bifunctional organoclay, has been prepared. It is able not only to sorb organophosphate pesticides, but also to catalyze their hydrolysis, and thereby detoxify them. The bifunctional organoclay prepared in this study is based on sodium montmorillonite, in which the inorganic counter ions are replaced by N-decyl-N,N-dimethyl-N-(2-aminoethyl) ammonium (DDMAEA). The detoxifying capacity of this organoclay for two organophosphate pesticides, methyl parathion [O,O-dimethyl O-(p-nitrophenyl) thionophosphate] and tetrachlorvinphos [2-chloro-1-(2,4,5-trichlorophenyl)ethenyl dimethyl phosphate], was demonstrated. It was shown that although the sorption of these pesticides on the bifunctional organoclay is very similar to that on N-decyl-N,N,N-trimethyl ammonium (DTMA) organoclay (the corresponding nonbifunctional organoclay), the hydrolysis of these pesticides is substantially enhanced only by the bifunctional organoclay. The half-life for the hydrolysis of the investigated pesticides in the presence of the bifunctional organoclay is about 12 times less than for their spontaneous hydrolysis, and the enhancement is even more pronounced relative to the hydrolysis of these pesticides in the presence of the DTMA organoclay (which actually inhibits their hydrolysis). Based on kinetic measurements, the pK(a) of the ethylamino group of the bifunctional organoclay was estimated to be around 9.0. It is postulated that the catalytic effect of the bifunctional organoclay can be attributed to a nucleophilic attack of the unprotonated ethylamino group of the organoclay on the organophosphate ester. PMID- 15356257 TI - Estimating bulk density in vertically exposed stoney alluvium using a modified excavation method. AB - Despite many decades of education and refining land-use practices, accelerated stream bank erosion is still prevalent in the United States. Eroding stream banks produce a sediment load to the riverine system and can cause reduced water quality as a result of increased suspended sediment. As total maximum daily loads (TMDLs) for water bodies impaired by turbidity or suspended sediments become more numerous, a simple, in situ field technique will be needed to estimate the bulk density of readily erodible stream bank material so that reasonably accurate sediment loading rates can be estimated. In this study, the excavation/polyurethane-foam technique for estimating total bulk density was applied to vertically exposed alluvium with high coarse-fragment content. Though not previously attempted in vertically exposed alluvium with high coarse-fragment content, the excavation/polyurethane-foam technique appears to provide a reasonably accurate estimate of the total and soil (<2-mm size fraction) bulk density from vertically exposed, alluvial deposits with high coarse-fragment content (i.e., >70%) along eroding stream banks. Obtaining bulk density estimates using this method would facilitate calculation of sediment loading rates to riverine systems with actual field data. PMID- 15356258 TI - Adsorption of trace metals on glass fiber filters. AB - Filters, containing glass-fiber (GF) filter material, are commonly used as the primary filter or as the prefilter in sampling natural waters and laboratory experiments with high concentrations of suspended solids. We observed that GF filter material removed substantial quantities of trace metals from solutions of low ionic strength at near neutral and slightly acidic pH. The GF material sorbed essentially all Pb and Ag from 5-mL aliquots of solutions containing 0.054 and 0.093 mM, respectively. Somewhat less Ni was sorbed from a 0.099 mM solution. This material retained about 43 micromol of Ag per gram of GF material (4600 microg/g). The Ag and Ni sorption was highest at low KNO(3) concentrations (as background electrolyte) and decreased to a constant concentration of sorbed metal at approximately 10 mM KNO(3). Glass-fiber filter material should only be used with careful testing for the elements of concern under conditions that closely match expected environmental or experimental conditions. PMID- 15356259 TI - Formation of nuclear splicing factor compartments is independent of lamins A/C. AB - Nuclear lamins are major architectural elements of the mammalian cell nucleus, and they have been implicated in the functional organization of the nuclear interior, possibly by providing structural support for nuclear compartments. Colocalization studies have suggested a structural role for lamins in the formation and maintenance of pre-mRNA splicing factor compartments. Here, we have directly tested this hypothesis by analysis of embryonic fibroblasts from knock out mice lacking A- and C-type lamins. We show that the morphology and cellular properties of splicing factor compartments are independent of A- and C-type lamins. Genetic loss of lamins A/C has no effect on the cellular distribution of several pre-mRNA splicing factors and does not affect the compartment morphology as examined by light and electron microscopy. The association of splicing factors with the nuclear matrix fraction persists in the absence of lamins A/C. Live cell microscopy demonstrates that the intranuclear positional stability of splicing factor compartments is maintained and that the exchange dynamics of SF2/ASF between the compartments and the nucleoplasm is not affected by loss of lamin A/C. Our results demonstrate that formation and maintenance of intranuclear splicing factor compartments is independent of lamins A/C, and they argue against an essential structural role of lamins A/C in splicing factor compartment morphology. PMID- 15356260 TI - Interactions among Rax1p, Rax2p, Bud8p, and Bud9p in marking cortical sites for bipolar bud-site selection in yeast. AB - In the budding yeast Saccharomyces cerevisiae, selection of the bud site determines the axis of polarized cell growth and eventual oriented cell division. Bud sites are selected in specific patterns depending on cell type. These patterns appear to depend on distinct types of marker proteins in the cell cortex; in particular, the bipolar budding of diploid cells depends on persistent landmarks at the birth-scar-distal and -proximal poles that involve the proteins Bud8p and Bud9p, respectively. Rax1p and Rax2p also appear to function specifically in bipolar budding, and we report here a further characterization of these proteins and of their interactions with Bud8p and Bud9p. Rax1p and Rax2p both appear to be integral membrane proteins. Although commonly used programs predict different topologies for Rax2p, glycosylation studies indicate that it has a type I orientation, with its long N-terminal domain in the extracytoplasmic space. Analysis of rax1 and rax2 mutant budding patterns indicates that both proteins are involved in selecting bud sites at both the distal and proximal poles of daughter cells as well as near previously used division sites on mother cells. Consistent with this, GFP-tagged Rax1p and Rax2p were both observed at the distal pole as well as at the division site on both mother and daughter cells; localization to the division sites was persistent through multiple cell cycles. Localization of Rax1p and Rax2p was interdependent, and biochemical studies showed that these proteins could be copurified from yeast. Bud8p and Bud9p could also be copurified with Rax1p, and localization studies provided further evidence of interactions. Localization of Rax1p and Rax2p to the bud tip and distal pole depended on Bud8p, and normal localization of Bud8p was partially dependent on Rax1p and Rax2p. Although localization of Rax1p and Rax2p to the division site did not appear to depend on Bud9p, normal localization of Bud9p appeared largely or entirely dependent on Rax1p and Rax2p. Taken together, the results indicate that Rax1p and Rax2p interact closely with each other and with Bud8p and Bud9p in the establishment and/or maintenance of the cortical landmarks for bipolar budding. PMID- 15356261 TI - Megator, an essential coiled-coil protein that localizes to the putative spindle matrix during mitosis in Drosophila. AB - We have used immunocytochemistry and cross-immunoprecipitation analysis to demonstrate that Megator (Bx34 antigen), a Tpr ortholog in Drosophila with an extended coiled-coil domain, colocalizes with the putative spindle matrix proteins Skeletor and Chromator during mitosis. Analysis of P-element mutations in the Megator locus showed that Megator is an essential protein. During interphase Megator is localized to the nuclear rim and occupies the intranuclear space surrounding the chromosomes. However, during mitosis Megator reorganizes and aligns together with Skeletor and Chromator into a fusiform spindle structure. The Megator metaphase spindle persists in the absence of microtubule spindles, strongly implying that the existence of the Megator-defined spindle does not require polymerized microtubules. Deletion construct analysis in S2 cells indicates that the COOH-terminal part of Megator without the coiled-coil region was sufficient for both nuclear as well as spindle localization. In contrast, the NH2-terminal coiled-coil region remains in the cytoplasm; however, we show that it is capable of assembling into spherical structures. On the basis of these findings we propose that the COOH-terminal domain of Megator functions as a targeting and localization domain, whereas the NH2-terminal domain is responsible for forming polymers that may serve as a structural basis for the putative spindle matrix complex. PMID- 15356262 TI - Modulation of the biological activity of a tobacco LTP1 by lipid complexation. AB - Plant lipid transfer proteins (LTPs) are small, cysteine-rich proteins secreted into the extracellular space. They belong to the pathogenesis-related proteins (PR-14) family and are believed to be involved in several physiological processes including plant disease resistance, although their precise biological function is still unknown. Here, we show that a recombinant tobacco LTP1 is able to load fatty acids and jasmonic acid. This LTP1 binds to specific plasma membrane sites, previously characterized as elicitin receptors, and is shown to be involved in the activation of plant defense. The biological properties of this LTP1 were compared with those of LTP1-linolenic and LTP1-jasmonic acid complexes. The binding curve of the LTP1-linolenic acid complex to purified tobacco plasma membranes is comparable to the curve obtained with LTP1. In contrast, the LTP1 jasmonic acid complex shows a strongly increased interaction with the plasma membrane receptors. Treatment of tobacco plants with LTP1-jasmonic acid resulted in an enhancement of resistance toward Phytophthora parasitica. These effects were absent upon treatment with LTP1 or jasmonic acid alone. This work presents the first evidence for a biological activity of a LTP1 and points out the crucial role of protein-specific lipophilic ligand interaction in the modulation of the protein activity. PMID- 15356263 TI - The small subunit processome is required for cell cycle progression at G1. AB - Without ribosome biogenesis, translation of mRNA into protein ceases and cellular growth stops. We asked whether ribosome biogenesis is cell cycle regulated in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe, and we determined that it is not regulated in the same manner as in metazoan cells. We therefore turned our attention to cellular sensors that relay cell size information via ribosome biogenesis. Our results indicate that the small subunit (SSU) processome, a complex consisting of 40 proteins and the U3 small nucleolar RNA necessary for ribosome biogenesis, is not mitotically regulated. Furthermore, Nan1/Utp17, an SSU processome protein, does not provide a link between ribosome biogenesis and cell growth. However, when individual SSU processome proteins are depleted, cells arrest in the G1 phase of the cell cycle. This arrest was further supported by the lack of staining for proteins expressed in post-G1. Similarly, synchronized cells depleted of SSU processome proteins did not enter G2. This suggests that when ribosomes are no longer made, the cells stall in the G1. Therefore, yeast cells must grow to a critical size, which is dependent upon having a sufficient number of ribosomes during the G1 phase of the cell cycle, before cell division can occur. PMID- 15356264 TI - Role of Vma21p in assembly and transport of the yeast vacuolar ATPase. AB - The Saccharomyces cerevisiae vacuolar H+-ATPase (V-ATPase) is a multisubunit complex composed of a peripheral membrane sector (V1) responsible for ATP hydrolysis and an integral membrane sector (V0) required for proton translocation. Biogenesis of V0 requires an endoplasmic reticulum (ER)-localized accessory factor, Vma21p. We found that in vma21Delta cells, the major proteolipid subunit of V0 failed to interact with the 100-kDa V0 subunit, Vph1p, indicating that Vma21p is necessary for V0 assembly. Immunoprecipitation of Vma21p from wild-type membranes resulted in coimmunoprecipitation of all five V0 subunits. Analysis of vmaDelta strains showed that binding of V0 subunits to Vma21p was mediated by the proteolipid subunit Vma11p. Although Vma21p/proteolipid interactions were independent of Vph1p, Vma21p/Vph1p association was dependent on all other V0 subunits, indicating that assembly of V0 occurs in a defined sequence, with Vph1p recruitment into a Vma21p/proteolipid/Vma6p complex representing the final step. An in vitro assay for ER export was used to demonstrate preferential packaging of the fully assembled Vma21p/proteolipid/Vma6p/Vph1p complex into COPII-coated transport vesicles. Pulse-chase experiments showed that the interaction between Vma21p and V0 was transient and that Vma21p/V0 dissociation was concomitant with V0/V1 assembly. Blocking ER export in vivo stabilized the interaction between Vma21p and V0 and abrogated assembly of V0/V1. Although a Vma21p mutant lacking an ER retrieval signal remained associated with V0 in the vacuole, this interaction did not affect the assembly of vacuolar V0/V1 complexes. We conclude that Vma21p is not involved in regulating the interaction between V0 and V1 sectors, but that it has a crucial role in coordinating the assembly of V0 subunits and in escorting the assembled V0 complex into ER-derived transport vesicles. PMID- 15356265 TI - A high-affinity interaction with ADP-actin monomers underlies the mechanism and in vivo function of Srv2/cyclase-associated protein. AB - Cyclase-associated protein (CAP), also called Srv2 in Saccharomyces cerevisiae, is a conserved actin monomer-binding protein that promotes cofilin-dependent actin turnover in vitro and in vivo. However, little is known about the mechanism underlying this function. Here, we show that S. cerevisiae CAP binds with strong preference to ADP-G-actin (Kd 0.02 microM) compared with ATP-G-actin (Kd 1.9 microM) and competes directly with cofilin for binding ADP-G-actin. Further, CAP blocks actin monomer addition specifically to barbed ends of filaments, in contrast to profilin, which blocks monomer addition to pointed ends of filaments. The actin-binding domain of CAP is more extensive than previously suggested and includes a recently solved beta-sheet structure in the C-terminus of CAP and adjacent sequences. Using site-directed mutagenesis, we define evolutionarily conserved residues that mediate binding to ADP-G-actin and demonstrate that these activities are required for CAP function in vivo in directing actin organization and polarized cell growth. Together, our data suggest that in vivo CAP competes with cofilin for binding ADP-actin monomers, allows rapid nucleotide exchange to occur on actin, and then because of its 100-fold weaker binding affinity for ATP actin compared with ADP-actin, allows other cellular factors such as profilin to take the handoff of ATP-actin and facilitate barbed end assembly. PMID- 15356266 TI - Arf1p provides an unexpected link between COPI vesicles and mRNA in Saccharomyces cerevisiae. AB - The small GTPase Arf1p is involved in different cellular processes that require its accumulation at specific cellular locations. The recruitment of Arf1p to distinct points of action might be achieved by association of Arf1p with different proteins. To identify new interactors of Arf1p, we performed an affinity chromatography with GTP- or GDP-bound Arf1p proteins. A new interactor of Arf1p-GTP was identified as Pab1p, which binds to the polyA-tail of mRNAs. Pab1p was found to associate with purified COPI-coated vesicles generated from Golgi membranes in vitro. The stability of the Pab1p-Arf1p complex depends on the presence of mRNA. Both symmetrically distributed mRNAs as well as the asymmetrically localized ASH1 mRNA are found in association with Arf1p. Remarkably, Arf1p and Pab1p are both required to restrict ASH1 mRNA to the bud tip. Arf1p and coatomer play an unexpected role in localizing mRNA independent and downstream of the SHE machinery. Hereby acts the SHE machinery in long-range mRNA transport, whereas COPI vesicles could act as short-range and localization vehicles. The endoplasmic reticulum (ER)-Golgi shuttle might be involved in concentrating mRNA at the ER. PMID- 15356267 TI - Roles of the mammalian mitochondrial fission and fusion mediators Fis1, Drp1, and Opa1 in apoptosis. AB - During apoptosis, the mitochondrial network fragments. Using short hairpin RNAs for RNA interference, we manipulated the expression levels of the proteins hFis1, Drp1, and Opa1 that are involved in mitochondrial fission and fusion in mammalian cells, and we characterized their functions in mitochondrial morphology and apoptosis. Down-regulation of hFis1 powerfully inhibits cell death to an extent significantly greater than down-regulation of Drp1 and at a stage of apoptosis distinct from that induced by Drp1 inhibition. Cells depleted of Opa1 are extremely sensitive to exogenous apoptosis induction, and some die spontaneously by a process that requires hFis1 expression. Wild-type Opa1 may function normally as an antiapoptotic protein, keeping spontaneous apoptosis in check. However, if hFis1 is down-regulated, cells do not require Opa1 to prevent apoptosis, suggesting that Opa1 may be normally counteracting the proapoptotic action of hFis1. We also demonstrate in this study that mitochondrial fragmentation per se does not result in apoptosis. However, we provide further evidence that multiple components of the mitochondrial morphogenesis machinery can positively and negatively regulate apoptosis. PMID- 15356268 TI - GIPC recruits GAIP (RGS19) to attenuate dopamine D2 receptor signaling. AB - Pleiotropic G proteins are essential for the action of hormones and neurotransmitters and are activated by stimulation of G protein-coupled receptors (GPCR), which initiates heterotrimer dissociation of the G protein, exchange of GDP for GTP on its Galpha subunit and activation of effector proteins. Regulator of G protein signaling (RGS) proteins regulate this cascade and can be recruited to the membrane upon GPCR activation. Direct functional interaction between RGS and GPCR has been hypothesized. We show that recruitment of GAIP (RGS19) by the dopamine D2 receptor (D2R), a GPCR, required the scaffold protein GIPC (GAIP interacting protein, C terminus) and that all three were coexpressed in neurons and neuroendocrine cells. Dynamic translocation of GAIP to the plasma membrane and coassembly in a protein complex in which GIPC was a required component was dictated by D2R activation and physical interactions. In addition, two different D2R-mediated responses were regulated by the GTPase activity of GAIP at the level of the G protein coupling in a GIPC-dependent manner. Since GIPC exclusively interacted with GAIP and selectively with subsets of GPCR, this mechanism may serve to sort GPCR signaling in cells that usually express a large repertoire of GPCRs, G proteins, and RGS. PMID- 15356269 TI - Differential regulation of the TRAIL death receptors DR4 and DR5 by the signal recognition particle. AB - TRAIL (TNF-related apoptosis-inducing ligand) death receptors DR4 and DR5 facilitate the selective elimination of malignant cells through the induction of apoptosis. From previous studies the regulation of the DR4 and DR5 cell-death pathways appeared similar; nevertheless in this study we screened a library of small interfering RNA (siRNA) for genes, which when silenced, differentially affect DR4- vs. DR5-mediated apoptosis. These experiments revealed that expression of the signal recognition particle (SRP) complex is essential for apoptosis mediated by DR4, but not DR5. Selective diminution of SRP subunits by RNA interference resulted in a dramatic decrease in cell surface DR4 receptors that correlated with inhibition of DR4-dependent cell death. Conversely, SRP silencing had little influence on cell surface DR5 levels or DR5-mediated apoptosis. Although loss of SRP function in bacteria, yeast and protozoan parasites causes lethality or severe growth defects, we observed no overt phenotypes in the human cancer cells studied--even in stable cell lines with diminished expression of SRP components. The lack of severe phenotype after SRP depletion allowed us to delineate, for the first time, a mechanism for the differential regulation of the TRAIL death receptors DR4 and DR5--implicating the SRP complex as an essential component of the DR4 cell-death pathway. PMID- 15356270 TI - Echovirus 1 endocytosis into caveosomes requires lipid rafts, dynamin II, and signaling events. AB - Binding of echovirus 1 (EV1, a nonenveloped RNA virus) to the alpha2beta1 integrin on the cell surface is followed by endocytic internalization of the virus together with the receptor. Here, video-enhanced live microscopy revealed the rapid uptake of fluorescently labeled EV1 into mobile, intracellular structures, positive for green fluorescent protein-tagged caveolin-1. Partial colocalization of EV1 with SV40 (SV40) and cholera toxin, known to traffic via caveosomes, demonstrated that the vesicles were caveosomes. The initiation of EV1 infection was dependent on dynamin II, cholesterol, and protein phosphorylation events. Brefeldin A, a drug that prevents SV40 transport, blocked the EV1 infection cycle, whereas drugs that disrupt the cellular cytoskeleton had no effect. In situ hybridization revealed the localization of viral RNA with endocytosed viral capsid proteins in caveosomes before initiation of viral replication. Thus, both the internalization of EV1 to caveosomes and subsequent events differ clearly from caveolar endocytosis of SV40 because EV1 uptake is fast and independent of actin and EV1 is not sorted further to sER from caveosomes. These results shed further light on the cell entry of nonenveloped viral pathogens and illustrate the use of viruses as probes to dissect caveolin associated endocytic pathways. PMID- 15356271 TI - Susceptibility to apoptosis in insulin-like growth factor-I receptor-deficient brown adipocytes. AB - Fetal brown adipocytes are insulin-like growth factor-I (IGF-I) target cells. To assess the importance of the IGF-I receptor (IGF-IR) in brown adipocytes during fetal life, we have generated immortalized brown adipocyte cell lines from the IGF-IR(-/-) mice. Using this experimental model, we demonstrate that the lack of IGF-IR in fetal brown adipocytes increased the susceptibility to apoptosis induced by serum withdrawal. Culture of cells in the absence of serum and growth factors produced rapid DNA fragmentation (4 h) in IGF-IR(-/-) brown adipocytes, compared with the wild type (16 h). Consequently, cell viability was decreased more rapidly in fetal brown adipocytes in the absence of IGF-IR. Furthermore, caspase-3 activity was induced much earlier in cells lacking IGF-IR. At the molecular level, IGF-IR deficiency in fetal brown adipocytes altered the balance of the expression of several proapoptotic (Bcl-xS and Bim) and antiapoptotic (Bcl 2 and Bcl-xL) members of the Bcl-2 family. This imbalance was irreversible even though in IGF-IR-reconstituted cells. Likewise, cytosolic cytochrome c levels increased rapidly in IGF-IR-deficient cells compared with the wild type. A rapid entry of Foxo1 into the nucleus accompanied by a rapid exit from the cytosol and an earlier activation of caspase-8 were observed in brown adipocytes lacking IGF IR upon serum deprivation. Activation of caspase-8 was inhibited by 50% in both cell types by neutralizing anti-Fas-ligand antibody. Adenoviral infection of wild type brown adipocytes with constitutively active Foxol (ADA) increased the expression of antiapoptotic genes, decreased Bcl-xL and induced caspase-8 and -3 activities, with the final outcome of DNA fragmentation. Up-regulation of uncoupling protein-1 (UCP-1) expression in IGF-IR-deficient cells by transduction with PGC-1alpha or UCP-1 ameliorated caspase-3 activation, thereby retarding apoptosis. Finally, insulin treatment prevented apoptosis in both cell types. However, the survival effect of insulin on IGF-IR(-/-) brown adipocytes was elicited even in the absence of phosphatidylinositol 3-kinase/Akt signaling. Thus, our results demonstrate for the first time the unique role of IGF-IR in maintaining the balance of death and survival in fetal brown adipocytes. PMID- 15356273 TI - Three-dimensional window analysis for detecting positive selection at structural regions of proteins. AB - Detection of natural selection operating at the amino acid sequence level is important in the study of molecular evolution. Single-site analysis and one dimensional window analysis can be used to detect selection when the biological functions of amino acid sites are unknown. Single-site analysis is useful when selection operates more or less constantly over evolutionary time, but less so when selection operates temporarily. One-dimensional window analysis is more sensitive than single-site analysis when the functions of amino acid sites in close proximity in the linear sequence are similar, although this is not always the case. Here I present a three-dimensional window analysis method for detecting selection given the three-dimensional structure of the protein of interest. In the three-dimensional structure, the window is defined as the sphere centered on the alpha-carbon of an amino acid site. The window size is the radius of the sphere. The sites whose alpha-carbons are included in the window are grouped for the neutrality test. The window is moved within the three-dimensional structure by sequentially moving the central site along the primary amino acid sequence. To detect positive selection, it may also be useful to group the surface-exposed sites in the window separately. Three-dimensional window analysis appears not only to be more sensitive than single-site analysis and one-dimensional window analysis but also to provide similar specificity for inferring positive selection in the analyses of the hemagglutinin and neuraminidase genes of human influenza A viruses. This method, however, may fail to detect selection when it operates only on a particular site, in which case single-site analysis may be preferred, although a large number of sequences is required. PMID- 15356272 TI - Cleavages within the prodomain direct intracellular trafficking and degradation of mature bone morphogenetic protein-4. AB - Pro bone morphogenetic protein-4 (BMP-4) is initially cleaved at a consensus furin motif adjacent to the mature ligand domain (the S1 site), and this allows for subsequent cleavage at an upstream motif (the S2 site). Previous studies have shown that S2 cleavage regulates the activity and signaling range of mature BMP 4, but the mechanism by which this occurs is unknown. Here, we show that the pro- and mature domains of BMP-4 remain noncovalently associated after S1 cleavage, generating a complex that is targeted for rapid degradation. Degradation requires lysosomal and proteosomal function and is enhanced by interaction with heparin sulfate proteoglycans. Subsequent cleavage at the S2 site liberates mature BMP-4 from the prodomain, thereby stabilizing the protein. We also show that cleavage at the S2, but not the S1 site, is enhanced at reduced pH, consistent with the possibility that the two cleavages occur in distinct subcellular compartments. Based on these results, we propose a model for how cleavage at the upstream site regulates the activity and signaling range of mature BMP-4 after it has been released from the prodomain. PMID- 15356274 TI - Unique origin and lateral transfer of prokaryotic chlorophyll-b and chlorophyll-d light-harvesting systems. AB - pcb genes, encoding proteins binding light-harvesting chlorophylls, were cloned and sequenced from the Chl d-containing cyanobacterium, Acaryochloris marina, and the Chl b-containing cyanobacterium, Prochloron didemni. Both organisms contained two tandem pcb genes. Peptide fingerprinting confirmed the expression of one of the A. marina pcb genes. Phylogenetic tree reconstruction using distance-matrix and maximum-likelihood methods indicated a single origin of the pcb gene family, whether occurring in Chl b-containing or Chl d-containing organisms. This may indicate widespread lateral transfer of the Pcb protein-based light-harvesting system. PMID- 15356275 TI - Structural and evolutionary analyses of the Ty3/gypsy group of LTR retrotransposons in the genome of Anopheles gambiae. AB - The recent availability of the genome of Anopheles gambiae offers an extraordinary opportunity for comparative studies of the diversity of transposable elements (TEs) and their evolutionary dynamics between two related species, taking advantage of the existing information from Drosophila melanogaster. To this goal, we screened the genome of A. gambiae for elements belonging to the Ty3/gypsy group of long-terminal repeat (LTR) retrotransposons. The A. gambiae genome displays a rich diversity of LTR retrotransposons, clearly greater than D. melanogaster. We have characterized in detail 63 families, belonging to five of the nine main lineages of the Ty3/gypsy group. The Mag lineage is the most diverse and abundant, with more than 30 families. In sharp contrast with this finding, a single family belonging to this lineage has been found in D. melanogaster, here reported for the first time in the literature, most probably consisting of old inactive elements. The CsRn1 lineage is also abundant in A. gambiae but almost absent from D. melanogaster. Conversely, the Osvaldo lineage has been detected in Drosophila but not in Anopheles. Comparison of structural characteristics of different families led to the identification of several lineage-specific features such as the primer-binding site (PBS), the gag pol translational recoding signal (TRS), which is extraordinarily diverse within the Ty3/gypsy retrotransposons of A. gambiae, or the presence/absence of specific amino acid motifs. Interestingly, some of these characteristics, although in general well conserved within lineages, may have evolved independently in particular branches of the phylogenetic tree. We also show evidence of recent activity for around 75% of the families. Nevertheless, almost all families contain a high proportion of degenerate members and solitary LTRs (solo LTRs), indicative of a lower turnover rate of retrotransposons belonging to the Ty3/gypsy group in A. gambiae than in D. melanogaster. Finally, we have detected significant overrepresentations of insertions on the X chromosome versus autosomes and of putatively active insertions on euchromatin versus heterochromatin. PMID- 15356276 TI - Disentangling the effects of demography and selection in human history. AB - Demographic events affect all genes in a genome, whereas natural selection has only local effects. Using publicly available data from 151 loci sequenced in both European-American and African-American populations, we attempt to distinguish the effects of demography and selection. To analyze large sets of population genetic data such as this one, we introduce "Perlymorphism," a Unix-based suite of analysis tools. Our analyses show that the demographic histories of human populations can account for a large proportion of effects on the level and frequency of variation across the genome. The African-American population shows both a higher level of nucleotide diversity and more negative values of Tajima's D statistic than does a European-American population. Using coalescent simulations, we show that the significantly negative values of the D statistic in African-Americans and the positive values in European-Americans are well explained by relatively simple models of population admixture and bottleneck, respectively. Working within these nonequilibrium frameworks, we are still able to show deviations from neutral expectations at a number of loci, including ABO and TRPV6. In addition, we show that the frequency spectrum of mutations- corrected for levels of polymorphism--is correlated with recombination rate only in European-Americans. These results are consistent with repeated selective sweeps in non-African populations, in agreement with recent reports using microsatellite data. PMID- 15356277 TI - Integrating the universal metabolism into a phylogenetic analysis. AB - The darwinian concept of "descent with modification" applies to metabolic pathways: pathways sharing similarities must have inherited them from an exclusive, hypothetical ancestral pathway. Comparative anatomy of biochemical pathways is performed using five criteria of homology. Primary homologies of "type I" were defined as several pathways sharing the same enzyme with high specificity for its substrate. Primary homologies of "type II" were defined as the sharing of similar enzymatic functions, cofactors, functional family, or recurrence of a set of reactions. Standard cladistic analysis is used to infer the evolutionary history of metabolic development and the relative ordering of biochemical reactions through time, from a single matrix integrating the whole basic universal metabolism. The cladogram shows that the earliest pathways to emerge are metabolism of amino acids of groups I and II (Asp, Asn, Glu, and Gln). The earliest enzymatic functions are mostly linked to amino acid catabolism: deamination, transamination, and decarboxylation. For some amino acids, catabolism and biosynthesis occur at the same time (Asp, Glu, Lys, and Met). Catabolism precedes anabolism for Asn, Gln, Arg, Trp, His, Tyr, and Phe, and anabolism precedes catabolism for Pro, Ala, Leu, Val, Ile, Cys, Gly, Ser, and Thr. The urea cycle evolves from arginine synthesis. Metabolism of fatty acids and sugars develops after the full development of metabolism of amino acids of groups I and II, and they are associated with the anabolism of amino acids of groups III and IV. Syntheses of aromatic amino acids are branched within sugar metabolism. The Krebs cycle occurs relatively late after the setting of metabolism of amino acids of groups I and II. One portion of the Krebs cycle has a catabolic origin, whereas the other portion has an anabolic origin in pathways of amino acids of groups III and IV. It is not possible to order glycolysis and gluconeogenesis with regard to the Krebs cycle, as they all belong to "period 6." Pentose-phosphate and Calvin cycles are later (periods 7 and 8, respectively). Cladistic analysis of the structure of biochemical pathways makes hypotheses in biochemical evolution explicit and parsimonious. PMID- 15356278 TI - Gene transfers from nanoarchaeota to an ancestor of diplomonads and parabasalids. AB - Rare evolutionary events, such as lateral gene transfers and gene fusions, may be useful to pinpoint, and correlate the timing of, key branches across the tree of life. For example, the shared possession of a transferred gene indicates a phylogenetic relationship among organismal lineages by virtue of their shared common ancestral recipient. Here, we present phylogenetic analyses of prolyl-tRNA and alanyl-tRNA synthetase genes that indicate lateral gene transfer events to an ancestor of the diplomonads and parabasalids from lineages more closely related to the newly discovered archaeal hyperthermophile Nanoarchaeum equitans (Nanoarchaeota) than to Crenarchaeota or Euryarchaeota. The support for this scenario is strong from all applied phylogenetic methods for the alanyl-tRNA sequences, whereas the phylogenetic analyses of the prolyl-tRNA sequences show some disagreements between methods, indicating that the donor lineage cannot be identified with a high degree of certainty. However, in both trees, the diplomonads and parabasalids branch together within the Archaea, strongly suggesting that these two groups of unicellular eukaryotes, often regarded as the two earliest independent offshoots of the eukaryotic lineage, share a common ancestor to the exclusion of the eukaryotic root. Unfortunately, the phylogenetic analyses of these two aminoacyl-tRNA synthetase genes are inconclusive regarding the position of the diplomonad/parabasalid group within the eukaryotes. Our results also show that the lineage leading to Nanoarchaeota branched off from Euryarchaeota and Crenarchaeota before the divergence of diplomonads and parabasalids, that this unexplored archaeal diversity, currently only represented by the hyperthermophilic organism Nanoarchaeum equitans, may include members living in close proximity to mesophilic eukaryotes, and that the presence of split genes in the Nanoarchaeum genome is a derived feature. PMID- 15356279 TI - Phylogenetic mapping of intron positions: a case study of translation initiation factor eIF2gamma. AB - Eukaryotic translation initiation factor 2 (eIF2) is a G protein that delivers the methionyl initiator tRNA to the small ribosomal subunit and releases it upon GTP hydrolysis after the recognition of the initiation codon. eIF2 is composed of three subunits, alpha, beta, and gamma. Subunit gamma shows the strongest conservation, and it confers both tRNA and GTP/GDP binding. Using intron positioning and protein sequence alignment, here we show that eIF2gamma is a suitable phylogenetic marker for eukaryotes. We determined or completed the sequences of 13 arthropod eIF2gamma genes. Analyzing the phylogenetic distribution of 52 different intron positions in 55 distantly related eIF2gamma genes, we identified ancient ones and shared derived introns in our data set. Obviously, intron positioning in eIF2gamma is evolutionarily conserved. However, there were episodes of complete and partial intron losses followed by intron gains. We identified 17 clusters of intron positions based on their distribution. The evolution of these clusters appears to be connected with preferred exon length and can be used to estimate the relative timing of intron gain because nearby precursor introns had to be erased from the gene before the new introns could be inserted. Moreover, we identified a putative case of intron sliding that constitutes a synapomorphic character state supporting monophyly of Coleoptera, Lepidoptera, and Diptera excluding Hymenoptera. We also performed tree reconstructions using the eIF2gamma protein sequences and intron positioning as phylogenetic information. Our results support the monophyly of Viridoplantae, Ascomycota, Homobasidiomyceta, and Apicomplexa. PMID- 15356280 TI - Phylogenetic analysis of the ING family of PHD finger proteins. AB - Since the discovery of ING1 class II tumor suppressors in 1996, five different ING genes (ING1 to ING5) encoding proteins with highly conserved plant homeodomain (PHD) motifs and several splicing isoforms of the ING1 and ING2 gene have been identified. The ING family functions in DNA repair and apoptosis in response to UV damage through binding to proliferating cell nuclear antigen (PCNA); chromatin remodeling and regulation of gene expression through regulating and/or targeting histone acetyltransferase/deacetylase (HAT/HDAC) activities; binding targets of rare phosphatidylinositol phosphates (PtdInsPs) that function in DNA damage-initiated stress signaling; and regulating brain tumor angiogenesis through transcriptional repression of NF-KB-responsive genes. To elucidate the evolutionary history of ING proteins and summarize what is known about regions highly conserved in the ING family members, we have examined the sequences and phylogenetic relationships of ING proteins across taxonomically diverse organisms. We have identified novel ING family members in rats, frogs, fish, mosquitoes, fruit flies, worms, fungi, and plants. We have also clarified the naming and classification of ING proteins based on our phylogenetic analysis to allow better understanding of the ING protein family. Using sequence similarities, we have identified novel regions and motifs of unknown function that are conserved across family members. An evolutionary history for the ING family of PHD finger proteins is presented that indicates that five ING genes are present in vertebrates. Three of these may be paralogs of ING genes found in arthropods, whereas nematodes, fungi, and green plants contain ING genes that have general features of the vertebrate ING family. PMID- 15356281 TI - Comparative methods for the analysis of gene-expression evolution: an example using yeast functional genomic data. AB - Understanding the evolution of gene function is a primary challenge of modern evolutionary biology. Despite an expanding database from genomic and developmental studies, we are lacking quantitative methods for analyzing the evolution of some important measures of gene function, such as gene-expression patterns. Here, we introduce phylogenetic comparative methods to compare different models of gene-expression evolution in a maximum-likelihood framework. We find that expression of duplicated genes has evolved according to a nonphylogenetic model, where closely related genes are no more likely than more distantly related genes to share common expression patterns. These results are consistent with previous studies that found rapid evolution of gene expression during the history of yeast. The comparative methods presented here are general enough to test a wide range of evolutionary hypotheses using genomic-scale data from any organism. PMID- 15356282 TI - A globin gene of ancient evolutionary origin in lower vertebrates: evidence for two distinct globin families in animals. AB - Hemoglobin, myoglobin, neuroglobin, and cytoglobin are four types of vertebrate globins with distinct tissue distributions and functions. Here, we report the identification of a fifth and novel globin gene from fish and amphibians, which has apparently been lost in the evolution of higher vertebrates (Amniota). Because its function is presently unknown, we tentatively call it globin X (GbX). Globin X sequences were obtained from three fish species, the zebrafish Danio rerio, the goldfish Carassius auratus, and the pufferfish Tetraodon nigroviridis, and the clawed frog Silurana tropicalis. Globin X sequences are distinct from vertebrate hemoglobins, myoglobins, neuroglobins, and cytoglobins. Globin X displays the highest identity scores with neuroglobin (approximately 26% to 35%), although it is not a neuronal protein, as revealed by RT-PCR experiments on goldfish RNA from various tissues. The distal ligand-binding and the proximal heme-binding histidines (E7 and F8), as well as the conserved phenylalanine CD1 are present in the globin X sequences, but because of extensions at the N terminal and C-terminal, the globin X proteins are longer than the typical eight alpha-helical globins and comprise about 200 amino acids. In addition to the conserved globin introns at helix positions B12.2 and G7.0, the globin X genes contain two introns in E10.2 and H10.0. The intron in E10.2 is shifted by 1 bp in respect to the vertebrate neuroglobin gene (E11.0), providing possible evidence for an intron sliding event. Phylogenetic analyses confirm an ancient evolutionary relationship of globin X with neuroglobin and suggest the existence of two distinct globin types in the last common ancestor of Protostomia and Deuterostomia. PMID- 15356283 TI - Ancestors of trans-splicing mitochondrial introns support serial sister group relationships of hornworts and mosses with vascular plants. AB - Some group II introns in the organelle genomes of plants and algae are disrupted and require trans-splicing of the affected exons from independent transcripts. A peculiar mitochondrial nad5 gene structure is universally conserved in flowering plants where two trans-splicing introns frame a tiny exon of only 22 nucleotides, and two additional conventional group II introns interrupt the nad5 reading frame at other sites. These four introns are absent in the liverwort Marchantia polymorpha, which carries a group I intron at an unrelated site in nad5. To determine how intron gains and losses have sculptured mitochondrial gene structures in early land-plant evolution, we have investigated the full nad5 gene structures in the three bryophyte classes and the fern Asplenium nidus. We find the single Marchantia group I intron nad5i753 present as the only intervening sequence in both closely (Corsinia and Monoclea) and distantly related (Noteroclada, Bazzania, and Haplomitrium) liverwort genera. In a taxonomically wide spectrum of mosses (Sphagnum, Encalypta, Timmia, Ulota, and Rhacocarpus); however, we additionally identify the angiosperm-type group II introns nad5i230 and nad5i1455. The latter is a cis-arranged homolog to one of the two angiosperm trans-splicing introns, notably the first of its kind in mosses. In the hornwort Anthoceros, the "moss and liverwort-type" group I intron nad5i753 is absent, and, besides nad5i230 and nad5i1455, intron nad5i1477 is present as the second ancestral group II intron which has evolved into a trans-splicing arrangement in angiosperms. The influence of highly frequent RNA editing, most notably in the genera Haplomitrium, Anthoceros, and Asplenium, on phylogenetic tree construction is investigated and discussed. Taken together, the data (1) support a sister group relationship of liverworts as a whole to all other embryophytes, (2) indicate loss of a group I and serial entries of group II introns in the nad5 gene during early evolution of the nonliverwort lineage, and (3) propose a placement of hornworts as sister group to tracheophytes. PMID- 15356284 TI - Selective expansion of the newly evolved genomic variants of retrotransposon 1731 in the Drosophila genomes. AB - The structural variants of the regulatory and coding regions of the LTR retrotransposon 1731 are described. Two classes of genomic copies of retrotransposon 1731, with and without frameshifting strategy to express Gag and Pol proteins, were earlier revealed in the D. melanogaster genome. Copies without frameshifting are shown to be evolved from an ancient variant with frameshifting and are widespread in the genomes of the melanogaster complex species. Position of a rare codon responsible for ribosome pausing and efficient frameshifting is identified. Two structural variants of 1731 LTRs were detected in the melanogaster complex species: the predominant structural variant A1A2 of 1731 LTR in the D. melanogaster, D. simulans, and D. sechellia genomes contains duplicated and diverged copies of 28 bp in the U3 region, whereas A1 variant lacking this duplication is expanded in the D. mauritiana genome. Selective expansion of the A1A2 variant was detected in the independently established D. melanogaster cell cultures. A1A2 variant is expressed in embryos, cell culture, and testes, whereas A1 is expressed only in testes of D. melanogaster. Relief of expression of the A1A2 but not A1 variant in the ovaries as a result of mutation in the RNA interference (RNAi) spn-E gene is shown. Thus, expansion of the recently evolved genomic variants of the LTR retrotransposon 1731 possessing a new translation strategy, duplication in the U3 region, and extended profile of expression is revealed. PMID- 15356285 TI - Evolution of the RNA polymerase B' subunit gene (rpoB') in Halobacteriales: a complementary molecular marker to the SSU rRNA gene. AB - Many prokaryotes have multiple ribosomal RNA operons. Generally, sequence differences between small subunit (SSU) rRNA genes are minor (<1%) and cause little concern for phylogenetic inference or environmental diversity studies. For Halobacteriales, an order of extremely halophilic, aerobic Archaea, within-genome SSU rRNA sequence divergence can exceed 5%, rendering phylogenetic assignment problematic. The RNA polymerase B' subunit gene (rpoB') is a single-copy conserved gene that may be an appropriate alternative phylogenetic marker for Halobacteriales. We sequenced a fragment of the rpoB' gene from 21 species, encompassing 15 genera of Halobacteriales. To examine the utility of rpoB' as a phylogenetic marker in Halobacteriales, we investigated three properties of rpoB' trees: the variation in resolution between trees inferred from the rpoB' DNA and RpoB' protein alignment, the degree of mutational saturation between taxa, and congruence with the SSU rRNA tree. The rpoB' DNA and protein trees were for the most part congruent and consistently recovered two well-supported monophyletic groups, the clade I and clade II haloarchaea, within a collection of less well resolved Halobacteriales lineages. A comparison of observed versus inferred numbers of substitution revealed mutational saturation in the rpoB' DNA data set, particularly between more distant species. Thus, the RpoB' protein sequence may be more reliable than the rpoB' DNA sequence for inferring Halobacteriales phylogeny. AU tests of tree selection indicated the trees inferred from rpoB' DNA and protein alignments were significantly incongruent with the SSU rRNA tree. We discuss possible explanations for this incongruence, including tree reconstruction artifact, differential paralog sampling, and lateral gene transfer. This is the first study of Halobacteriales evolution based on a marker other than the SSU rRNA gene. In addition, we present a valuable phylogenetic framework encompassing a broad diversity of Halobacteriales, in which novel sequences can be inserted for evolutionary, ecological, or taxonomic investigations. PMID- 15356286 TI - Multiplex mRNA assay using electrophoretic tags for high-throughput gene expression analysis. AB - We describe a novel multiplexing technology using a library of small fluorescent molecules, termed eTag molecules, to code and quantify mRNA targets. eTag molecules, which have the same fluorometric property, but distinct charge-to-mass ratios possess pre-defined electrophoretic characteristics and can be resolved using capillary electrophoresis. Coupled with primary Invader mRNA assay, eTag molecules were applied to simultaneously quantify up to 44 mRNA targets. This multiplexing approach was validated by examining a panel of inflammation responsive genes in human umbilical vein endothelial cells stimulated with inflammatory cytokine interleukin 1beta. The laser-induced fluorescence detection and electrokinetic sample injection process in capillary electrophoresis allows sensitive quantification of thousands of copies of mRNA molecules in a reaction. The assay is precise, as evaluated by measuring qualified Z' factor, a dimensionless and simple characteristic for applications in high-throughput screening using mRNA assays. Our data demonstrate the synergy between the multiplexing capability of eTag molecules by sensitive capillary electrophoresis detection and the isothermal linear amplification characteristics of the Invader assay. eTag multiplex mRNA assay presents a unique platform for sensitive, high sample throughput and multiplex gene expression analysis. PMID- 15356287 TI - Whole-genome expression profiling through fragment display and combinatorial gene identification. AB - There is a growing demand for highly parallel gene expression analysis with whole genome coverage, high sensitivity and high accuracy. Open systems such as differential display are capable of analyzing most of the expressed genome but are not quantitative and generally require manual identification of differentially expressed genes by sequencing. Closed systems such as microarrays use gene-specific probes and are, therefore, limited to studying specific genes in well-characterized species. Here, we describe Tangerine, a PCR-based system that combines the scope and generality of open systems with a robust and immediate identification algorithm using publicly available sequence information. By combinatorial analysis of three independent and complete DNA indexing profiles, each displaying the complete set of expressed transcripts on capillary electrophoresis, the method allows transcripts to be simultaneously quantified and identified. The method is sensitive, accurate and reproducible, and is amenable to high-throughput automated operation. PMID- 15356288 TI - Rapid tagging of endogenous mouse genes by recombineering and ES cell complementation of tetraploid blastocysts. AB - The construction of knockin vectors designed to modify endogenous genes in embryonic stem (ES) cells and the generation of mice from these modified cells is time consuming. The timeline of an experiment from the conception of an idea to the availability of mature mice is at least 9 months. We describe a method in which this timeline is typically reduced to 3 months. Knockin vectors are rapidly constructed from bacterial artificial chromosome clones by recombineering followed by gap-repair (GR) rescue, and mice are rapidly derived by injecting genetically modified ES cells into tetraploid blastocysts. We also describe a tandem affinity purification (TAP)/floxed marker gene plasmid and a GR rescue plasmid that can be used to TAP tag any murine gene. The combination of recombineering and tetraploid blastocyst complementation provides a means for large-scale TAP tagging of mammalian genes. PMID- 15356289 TI - Clinical trial registration: a statement from the International Committee of Medical Journal Editors. PMID- 15356290 TI - Identifying DNA-binding proteins using structural motifs and the electrostatic potential. AB - Robust methods to detect DNA-binding proteins from structures of unknown function are important for structural biology. This paper describes a method for identifying such proteins that (i) have a solvent accessible structural motif necessary for DNA-binding and (ii) a positive electrostatic potential in the region of the binding region. We focus on three structural motifs: helix-turn helix (HTH), helix-hairpin-helix (HhH) and helix-loop-helix (HLH). We find that the combination of these variables detect 78% of proteins with an HTH motif, which is a substantial improvement over previous work based purely on structural templates and is comparable to more complex methods of identifying DNA-binding proteins. Similar true positive fractions are achieved for the HhH and HLH motifs. We see evidence of wide evolutionary diversity for DNA-binding proteins with an HTH motif, and much smaller diversity for those with an HhH or HLH motif. PMID- 15356291 TI - Functional interaction between peroxisome proliferator-activated receptors-alpha and Mef-2C on human carnitine palmitoyltransferase 1beta (CPT1beta) gene activation. AB - Muscle-type carnitine palmitoyltransferase 1 (CPT1beta) is considered to be the gene that controls fatty acid mitochondrial beta-oxidation. A functional peroxisome proliferator-activated receptor (PPAR) responsive element (PPRE) and a myocite-specific (MEF2) site that binds MEF2A and MEF2C in the promoter of this gene had been previously identified. We investigated the roles of the PPRE and the MEF2 binding sites and the potential interaction between PPARalpha and MEF2C regulating the CPT1beta gene promoter. Mutation analysis indicated that the MEF2 site contributed to the activation of the CPT1beta promoter by PPAR in C2C12 cells. The reporter construct containing the PPRE and the MEF2C site was synergistically activated by co-expression of PPAR, retinoid X receptor (RXR) and MEF2C in non-muscle cells. Moreover, protein-binding assays demonstrated that MEF2C and PPAR specifically bound to one another in vitro. Also for the synergistic activation of the CPT1beta gene promoter by MEF2C and PPARalpha RXRalpha, a precise arrangement of its binding sites was essential. PMID- 15356292 TI - The glioma-amplified sequence 41 gene (GAS41) is a direct Myb target gene. AB - The retroviral oncogene v-myb encodes a transcription factor (v-Myb) which transforms myelomonocytic cells in vivo and in vitro. It is thought that v-Myb exerts its biological effects by deregulating the expression of specific target genes, most of which are still unknown. The chicken glioma-amplified sequence 41 gene (GAS41) is located immediately downstream of the lysozyme gene, a known Myb regulated gene. The GAS41 promoter colocalizes with a CpG island which also functions as an origin of replication. Since the GAS41 promoter contains several potential Myb-binding sites (MBSs) we have investigated whether GAS41 is a v-Myb target gene. Our results show that the GAS41 gene is directly activated by a v Myb/estrogen receptor fusion protein. Furthermore, our studies reveal that the GAS41 promoter is stimulated by v-Myb in co-transfection experiments and that the DNA-binding activity of v-Myb is crucial for transactivation of the promoter. Electrophoretic mobility-shift assays (EMSA) indicate that several Myb-binding sites, residing approximately 250 bp upstream of the transcriptional start site, are bound by Myb in vitro. Furthermore, chromatin immunoprecipitation assays demonstrate that v-Myb is bound to the GAS41 promoter in vivo. Taken together these findings identify the GAS41 gene as a novel v-Myb target gene. We have also analysed the GAS41 replication origin in myelomonocytic cells and have failed to observe significant differences in origin activity in cells expressing or not expressing v-Myb. PMID- 15356293 TI - Polyadenylation-dependent screening assay for respiratory syncytial virus RNA transcriptase activity and identification of an inhibitor. AB - RNA-dependent RNA polymerase from respiratory syncytial virus (RSV) is a multi subunit ribonucleoprotein (RNP) complex that, in addition to synthesizing the full 15 222 nt viral genomic RNA, is able to synthesize all 10 viral mRNAs. We have prepared crude RNP from RSV-infected HEp-2 cells, based on a method previously used for Newcastle disease virus, and established a novel polyadenylation-dependent capture [poly(A) capture] assay to screen for potential inhibitors of RSV transcriptase activity. In this homogeneous assay, radiolabeled full-length polyadenylated mRNAs produced by the viral RNP are detected through capture on immobilized biotinylated oligo(dT) in a 96-well streptavidin-coated FlashPlate. Possible inhibitors identified with this assay could interfere at any step required for the production of complete RSV mRNAs, including transcription, polyadenylation and, potentially, co-transcriptional guanylylation. A specific inhibitor of RSV transcriptase with antiviral activity was identified through screening of this assay. PMID- 15356294 TI - Both KH and non-KH domain sequences are required for polyribosome association of Scp160p in yeast. AB - Scp160p is a 160 kDa RNA-binding protein in yeast previously demonstrated to associate with specific messages as an mRNP component of both soluble and membrane-bound polyribosomes. Although the vast majority of Scp160p sequence consists of 14 closely spaced KH domains, comparative sequence analyses also demonstrate the presence of a potential nuclear localization sequence located between KH domains 3 and 4, as well as a 110 amino acid non-KH N-terminal region that includes a potential nuclear export sequence (NES). As a step toward investigating the structure/function relationships of Scp160p, we generated two truncated alleles, FLAG.SCP160DeltaN1, encoding a protein product that lacks the first 74 amino acids, including the potential NES, and FLAG.SCP160DeltaC1, encoding a protein product that lacks the final KH domain (KH14). We report here that the N-truncated protein, expressed as a green fluorescent protein fusion in yeast, remains cytoplasmic, with no apparent nuclear accumulation. Biochemical studies further demonstrate that although the N-truncated protein remains competent to form RNPs, the C-truncated protein does not. Furthermore, polyribosome association is severely compromised for both truncated proteins. Perhaps most important, both truncated alleles appear only marginally functional in vivo, as demonstrated by the inability of each to complement scp160/eap1 synthetic lethality in a tester strain. Together, these data challenge the notion that Scp160p normally shuttles between the nucleus and cytoplasm, and further implicate polyribosome association as an essential component of Scp160p function in vivo. Finally, these data underscore the vital roles of both KH and non-KH domain sequences in Scp160p. PMID- 15356295 TI - Structural requirements for pre-microRNA binding and nuclear export by Exportin 5. AB - The biogenesis and function of mature human microRNAs is dependent on the nuclear export of pre-microRNA precursors by Exportin 5 (Exp5). The precursor for the human miR-30 microRNA, which is a 63 nt long RNA hairpin bearing a 2 nt 3' overhang, forms a specific complex with Exp5 and the Ran-GTP cofactor. Here, we have examined the structural requirements for pre-microRNA binding by Exp5. Our data indicate that pre-miR-30 binding requires an RNA stem of >16 bp and is facilitated by a 3' overhang. Although a blunt-ended derivative of the pre-miR-30 stem-loop remained capable of binding Exp5, 5' overhangs were inhibitory. miR-30 variants that had lost the ability to bind Exp5 effectively were not efficiently exported from the nucleus and were also expressed at reduced levels. Furthermore, formation of a pre-microRNA/Exp5/Ran-GTP complex inhibited exonucleolytic digestion of the pre-miRNA in vitro. Together, these data demonstrate that pre microRNA binding by Exp5 involves recognition of almost all of the RNA hairpin, with the exception of the terminal loop. Moreover, these results argue that Exp5 binding not only mediates pre-microRNA nuclear export but also prevents nuclear pre-microRNA degradation. PMID- 15356296 TI - Portrait of transcriptional responses to ultraviolet and ionizing radiation in human cells. AB - To understand the human response to DNA damage, we used microarrays to measure transcriptional responses of 10 000 genes to ionizing radiation (IR) and ultraviolet radiation (UV). To identify bona fide responses, we used cell lines from 15 individuals and a rigorous statistical method, Significance Analysis of Microarrays (SAM). By exploring how sample number affects SAM, we rendered a portrait of the human damage response with a degree of accuracy unmatched by previous studies. By showing how SAM can be used to estimate the total number of responsive genes, we discovered that 24% of all genes respond to IR and 32% respond to UV, although most responses were less than 2-fold. Many genes were involved in known damage-response pathways for cell cycling and proliferation, apoptosis, DNA repair or the stress response. However, the majority of genes were involved in unexpected pathways, with functions in signal transduction, RNA binding and editing, protein synthesis and degradation, energy metabolism, metabolism of macromolecular precursors, cell structure and adhesion, vesicle transport, or lysosomal metabolism. Although these functions were not previously associated with the damage response in mammals, many were conserved in yeast. These insights reveal new directions for studying the human response to DNA damage. PMID- 15356297 TI - The interactome as a tree--an attempt to visualize the protein-protein interaction network in yeast. AB - The refinement and high-throughput of protein interaction detection methods offer us a protein-protein interaction network in yeast. The challenge coming along with the network is to find better ways to make it accessible for biological investigation. Visualization would be helpful for extraction of meaningful biological information from the network. However, traditional ways of visualizing the network are unsuitable because of the large number of proteins. Here, we provide a simple but information-rich approach for visualization which integrates topological and biological information. In our method, the topological information such as quasi-cliques or spoke-like modules of the network is extracted into a clustering tree, where biological information spanning from protein functional annotation to expression profile correlations can be annotated onto the representation of it. We have developed a software named PINC based on our approach. Compared with previous clustering methods, our clustering method ADJW performs well both in retaining a meaningful image of the protein interaction network as well as in enriching the image with biological information, therefore is more suitable in visualization of the network. PMID- 15356298 TI - Over 20% of human transcripts might form sense-antisense pairs. AB - The major challenge to identifying natural sense- antisense (SA) transcripts from public databases is how to determine the correct orientation for an expressed sequence, especially an expressed sequence tag sequence. In this study, we established a set of very stringent criteria to identify the correct orientation of each human transcript. We used these orientation-reliable transcripts to create 26 741 transcription clusters in the human genome. Our analysis shows that 22% (5880) of the human transcription clusters form SA pairs, higher than any previous estimates. Our orientation-specific RT-PCR results along with the comparison of experimental data from previous studies confirm that our SA data set is reliable. This study not only demonstrates that our criteria for the prediction of SA transcripts are efficient, but also provides additional convincing data to support the view that antisense transcription is quite pervasive in the human genome. In-depth analyses show that SA transcripts have some significant differences compared with other types of transcripts, with regard to chromosomal distribution and Gene Ontology-annotated categories of physiological roles, functions and spatial localizations of gene products. PMID- 15356299 TI - Despite the odds - providing reproductive health care to Afghan women. PMID- 15356300 TI - A view from the periphery - health care in rural America. PMID- 15356301 TI - Factor VIII, D-Dimer, and thromboembolism in children. PMID- 15356302 TI - Drugs and the QT interval - caveat doctor. PMID- 15356303 TI - The effect of air pollution on lung development from 10 to 18 years of age. AB - BACKGROUND: Whether exposure to air pollution adversely affects the growth of lung function during the period of rapid lung development that occurs between the ages of 10 and 18 years is unknown. METHODS: In this prospective study, we recruited 1759 children (average age, 10 years) from schools in 12 southern California communities and measured lung function annually for eight years. The rate of attrition was approximately 10 percent per year. The communities represented a wide range of ambient exposures to ozone, acid vapor, nitrogen dioxide, and particulate matter. Linear regression was used to examine the relationship of air pollution to the forced expiratory volume in one second (FEV(1)) and other spirometric measures. RESULTS: Over the eight-year period, deficits in the growth of FEV(1) were associated with exposure to nitrogen dioxide (P=0.005), acid vapor (P=0.004), particulate matter with an aerodynamic diameter of less than 2.5 microm (PM(2.5)) (P=0.04), and elemental carbon (P=0.007), even after adjustment for several potential confounders and effect modifiers. Associations were also observed for other spirometric measures. Exposure to pollutants was associated with clinically and statistically significant deficits in the FEV(1) attained at the age of 18 years. For example, the estimated proportion of 18-year-old subjects with a low FEV(1) (defined as a ratio of observed to expected FEV(1) of less than 80 percent) was 4.9 times as great at the highest level of exposure to PM(2.5) as at the lowest level of exposure (7.9 percent vs. 1.6 percent, P=0.002). CONCLUSIONS: The results of this study indicate that current levels of air pollution have chronic, adverse effects on lung development in children from the age of 10 to 18 years, leading to clinically significant deficits in attained FEV(1) as children reach adulthood. PMID- 15356304 TI - Results of a home-based environmental intervention among urban children with asthma. AB - BACKGROUND: Children with asthma who live in the inner city are exposed to multiple indoor allergens and environmental tobacco smoke in their homes. Reductions in these triggers of asthma have been difficult to achieve and have seldom been associated with decreased morbidity from asthma. The objective of this study was to determine whether an environmental intervention tailored to each child's allergic sensitization and environmental risk factors could improve asthma-related outcomes. METHODS: We enrolled 937 children with atopic asthma (age, 5 to 11 years) in seven major U.S. cities in a randomized, controlled trial of an environmental intervention that lasted one year (intervention year) and included education and remediation for exposure to both allergens and environmental tobacco smoke. Home environmental exposures were assessed every six months, and asthma-related complications were assessed every two months during the intervention and for one year after the intervention. RESULTS: For every 2 week period, the intervention group had fewer days with symptoms than did the control group both during the intervention year (3.39 vs. 4.20 days, P<0.001) and the year afterward (2.62 vs. 3.21 days, P<0.001), as well as greater declines in the levels of allergens at home, such as Dermatophagoides farinae (Der f1) allergen in the bed (P<0.001) and on the bedroom floor (P=0.004), D. pteronyssinus in the bed (P=0.007), and cockroach allergen on the bedroom floor (P<0.001). Reductions in the levels of cockroach allergen and dust-mite allergen (Der f1) on the bedroom floor were significantly correlated with reduced complications of asthma (P<0.001). CONCLUSIONS: Among inner-city children with atopic asthma, an individualized, home-based, comprehensive environmental intervention decreases exposure to indoor allergens, including cockroach and dust mite allergens, resulting in reduced asthma-associated morbidity. PMID- 15356305 TI - Elevated plasma factor VIII and D-dimer levels as predictors of poor outcomes of thrombosis in children. AB - BACKGROUND: Elevated levels of plasma factor VIII and D-dimer predict recurrent venous thromboembolism in adults. We sought to determine whether an elevation of factor VIII, D-dimer, or both at diagnosis and persistence of the laboratory abnormality after three to six months of anticoagulant therapy correlate with poor outcomes of thrombosis in children. METHODS: We evaluated levels of factor VIII and D-dimer and additional components of an extensive laboratory thrombophilia (i.e., hypercoagulability) panel at the time of diagnosis in 144 children with a radiologically confirmed acute thrombotic event. All patients were treated initially with heparin and then with either warfarin or low molecular-weight heparin for at least three to six months, according to the current standard of care. Patients were examined at follow-up visits 3, 6, and 12 months after diagnosis and then annually, at which times testing was repeated in children with previously abnormal factor VIII and D-dimer test results and a uniform evaluation for the post-thrombotic syndrome was performed. RESULTS: Among 82 children for whom complete data were available regarding laboratory test results at diagnosis and thrombotic outcomes during follow-up, 67 percent had factor VIII levels above the cutoff value of 150 IU per deciliter, D-dimer levels above 500 ng per milliliter, or both at diagnosis, and at least one of the two laboratory values was persistently elevated in 43 percent of the 75 patients in whom testing was performed after three to six months of anticoagulant therapy. Fifty-one percent of the 82 patients had a poor outcome (i.e., a lack of thrombus resolution, recurrent thrombosis, or the post-thrombotic syndrome) during a median follow-up of 12 months (range, 3 months to 5 years). Elevated levels of factor VIII, D-dimer, or both at diagnosis were highly predictive of a poor outcome (odds ratio, 6.1; P=0.008), as was the persistence of at least one laboratory abnormality at three to six months (odds ratio, 4.7; P=0.002). The combination of a factor VIII level above 150 IU per deciliter and a D-dimer level above 500 ng per milliliter at diagnosis was 91 percent specific for a poor outcome, and after three to six months of standard anticoagulation, the combination was 88 percent specific. CONCLUSIONS: Elevated levels of plasma factor VIII, D-dimer, or both at diagnosis and a persistent elevation of at least one of these factors after standard-duration anticoagulant therapy predict a poor outcome in children with thrombosis. PMID- 15356306 TI - Oral erythromycin and the risk of sudden death from cardiac causes. AB - BACKGROUND: Oral erythromycin prolongs cardiac repolarization and is associated with case reports of torsades de pointes. Because erythromycin is extensively metabolized by cytochrome P-450 3A (CYP3A) isozymes, commonly used medications that inhibit the effects of CYP3A may increase plasma erythromycin concentrations, thereby increasing the risk of ventricular arrhythmias and sudden death. We studied the association between the use of erythromycin and the risk of sudden death from cardiac causes and whether this risk was increased with the concurrent use of strong inhibitors of CYP3A. METHODS: We studied a previously identified Tennessee Medicaid cohort that included 1,249,943 person-years of follow-up and 1476 cases of confirmed sudden death from cardiac causes. The CYP3A inhibitors used in the study were nitroimidazole antifungal agents, diltiazem, verapamil, and troleandomycin; each doubles, at least, the area under the time concentration curve for a CYP3A substrate. Amoxicillin, an antimicrobial agent with similar indications but which does not prolong cardiac repolarization, and former use of erythromycin also were studied, to assess possible confounding by indication. RESULTS: The multivariate adjusted rate of sudden death from cardiac causes among patients currently using erythromycin was twice as high (incidence rate ratio, 2.01; 95 percent confidence interval, 1.08 to 3.75; P=0.03) as that among those who had not used any of the study antibiotic medications. There was no significant increase in the risk of sudden death among former users of erythromycin (incidence-rate ratio, 0.89; 95 percent confidence interval, 0.72 to 1.09; P=0.26) or among those who were currently using amoxicillin (incidence-rate ratio, 1.18; 95 percent confidence interval, 0.59 to 2.36; P=0.65). The adjusted rate of sudden death from cardiac causes was five times as high (incidence-rate ratio, 5.35; 95 percent confidence interval, 1.72 to 16.64; P=0.004) among those who concurrently used CYP3A inhibitors and erythromycin as that among those who had used neither CYP3A inhibitors nor any of the study antibiotic medications. In contrast, there was no increase in the risk of sudden death among those who concurrently used amoxicillin and CYP3A inhibitors or those currently using any of the study antibiotic medications who had formerly used CYP3A inhibitors. CONCLUSIONS: The concurrent use of erythromycin and strong inhibitors of CYP3A should be avoided. PMID- 15356307 TI - Clinical practice. Diastolic heart failure. PMID- 15356308 TI - Thiazolidinediones. PMID- 15356309 TI - Images in clinical medicine. Gallstone ileus. PMID- 15356310 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 28-2004. Newborn twins with thrombocytopenia, coagulation defects, and hepatosplenomegaly. PMID- 15356311 TI - Air pollution and health - good news and bad. PMID- 15356312 TI - Allergen avoidance to reduce asthma-related morbidity. PMID- 15356313 TI - Class - the ignored determinant of the nation's health. PMID- 15356314 TI - Diastolic heart failure. PMID- 15356315 TI - Obesity and the metabolic syndrome in children and adolescents. PMID- 15356316 TI - Palliative care. PMID- 15356317 TI - Case 11-2004: a boy with rash, edema, and hypertension. PMID- 15356318 TI - Havana and the coma and death symposia. PMID- 15356319 TI - Images in clinical medicine. Marchiafava-Bignami disease. PMID- 15356321 TI - ARC3, a chloroplast division factor, is a chimera of prokaryotic FtsZ and part of eukaryotic phosphatidylinositol-4-phosphate 5-kinase. AB - The arc3 (accumulation and replication of chloroplast) mutant of Arabidopsis thaliana has a small number of abnormally large chloroplasts in the cell, suggesting that chloroplast division is arrested in the mutant and ARC3 has an important role in the initiation of chloroplast division. To elucidate the role of ARC3, first we identified the ARC3 gene, and determined the location of ARC3 protein during chloroplast division because the localization and spatial orientation of such division factors are vital for correct chloroplast division. Sequencing analysis showed that ARC3 was a fusion of the prokaryotic FtsZ and part of the eukaryotic phosphatidylinositol-4-phosphate 5-kinase (PIP5K) genes. The PIP5K-homologous region of ARC3 had no catalytic domain but a membrane occupation-and-recognition-nexus (MORN) repeat motif. Immunofluorescence microscopy, Western blotting analysis and in vitro chloroplast import and protease protection assays revealed that ARC3 protein was soluble, and located on the outer surface of the chloroplast in a ring-like structure at the early stage of chloroplast division. Prokaryotes have one FtsZ as a gene for division but have no ARC3 counterparts, the chimera of FtsZ and PIP5K, suggesting that the ARC3 gene might have been generated from FtsZ as another division factor during the evolution of chloroplast by endosymbiosis. PMID- 15356322 TI - Degradation of phytochrome interacting factor 3 in phytochrome-mediated light signaling. AB - Plant photoreceptors regulate various developmental processes. Among the photoreceptors, phytochromes, red and far-red light receptors, regulate light responses through many signaling components, including phytochrome-interacting proteins. The functional relationships among phytochromes and their interacting proteins, however, have not been clearly established. Here, we sought to identify a functional relationship between phytochromes and phytochrome interacting factor 3 (PIF3). We demonstrate that PIF3 is polyubiquitinated rapidly and subsequently degraded in PHYA and PHYB-mediated light signaling. We also show that the degradation of PIF3 is mediated by the 26S proteasome. Our data indicate that light-stimulated phytochromes cause the degradation of their interacting protein, PIF3, by the 26S proteasome. PMID- 15356323 TI - Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene. AB - Roots of Panax ginseng, one of the most famous medicinal plants, contain various phytosterols and bioactive triterpene saponins (ginsenosides). In P. ginseng, phytosterols and triterpenes share the common biosynthetic intermediate, squalene. Here, we investigate the regulatory role of Panax ginseng squalene synthase (PgSS1) on the biosynthesis of phytosterols and triterpene saponins. PgSS1 transcripts are expressed ubiquitously in the various plant tissues, but higher in shoot apex and root. The transcript levels of PgSS1 increased markedly in the adventitious roots during 12- to 96-h period after metyl jasmonate (MeJA) treatment; MeJA treatment induced the activation of the transcripts of squalene epoxidase (SE), beta-amyrin synthase (bAS), but not cycloartenol synthase (CAS). Unlike MeJA treatment, overexpression of PgSS1 in adventitious roots of transgenic P. ginseng was followed by the up-regulation of all the downstream genes tested, such as SE, bAS, and CAS. The enhanced activity of PgSS1 enzyme resulted in remarkable increase of phytosterols as well as ginsenoside contents. These results demonstrate that PgSS1 is a key regulatory enzyme not only for phytosterol but also for triterpene biosynthesis and overexpressing of PgSS1 confers the hyperproduction of triterpene saponins to P. ginseng. PMID- 15356324 TI - The virescent-2 mutation inhibits translation of plastid transcripts for the plastid genetic system at an early stage of chloroplast differentiation. AB - The rice virescent-2 mutant (v(2)) is temperature conditional and develops chlorotic, chloroplast-deficient leaves at the restrictive temperature. In the v(2) mutant, plastid-encoded proteins involved in photosynthesis and plastid transcriptional regulation were not detectable at any time during chloroplast differentiation. However, the plastid transcripts for these two classes of proteins behaved differently in the mutant, with those for the plastid transcription/translation apparatus accumulating to wild-type levels and those for photosynthetic apparatus being suppressed. Polysome analysis showed that translation of the plastid transcripts encoding the plastid transcription/translation apparatus was blocked at an early stage of chloroplast differentiation. Accumulation of transcripts of nuclear-encoded photosynthetic genes, such as cab and rbcS, was strongly suppressed in the mutant at later stages of chloroplast differentiation, whereas transcripts of genes for the plastid transcription apparatus, such as OsRpoTp and OsSIG2A, accumulated to abnormally high levels at these stages. These results suggest that activation of the plastid translation machinery at an early stage of chloroplast differentiation is important for triggering the transmission of information about plastid developmental state to the nucleus, which in turn is required for the induction of nuclear-encoded chloroplast proteins at later stages of chloroplast differentiation. PMID- 15356325 TI - The acceptor availability at photosystem I and ABA control nuclear expression of 2-Cys peroxiredoxin-A in Arabidopsis thaliana. AB - The redox-regulated 2-Cys peroxiredoxin-A (2CPA) promoter, which drives expression of a dominant chloroplast antioxidant enzyme, responds to signals originating from the photosynthetic electron transport downstream of PSI. Modulation of CO(2)- and NO(3)(-) -reduction rates in reporter gene plants expressing glucuronidase under control of the Arabidopsis thaliana 2CPA promoter revealed that promoter activity correlates with the availability of electron acceptors at PSI. The photosynthetic redox-regulation can be simulated by oxidant and antioxidant treatments. Inhibitor studies with PD98059 and staurosporine showed that a mitogen-activated protein kinase kinase transmits the oxidative response, while the antioxidant signal is transmitted by a serine/threonine kinase. Analysis of 2CPA promoter regulation in the abscisic acid (ABA) biosynthetic mutants aba2 and aba3 and the ABA-insensitive mutants abi1 and abi2 support a regulatory circuitry in which the redox signal cross-talks with the ABA signaling cascade downstream of ABI1 and ABI2. PMID- 15356326 TI - Overexpression of the Arabidopsis 14-3-3 protein GF14 lambda in cotton leads to a "stay-green" phenotype and improves stress tolerance under moderate drought conditions. AB - The Arabidopsis gene GF14 lambda that encodes a 14-3-3 protein was introduced into cotton plants to explore the physiological roles that GF14 lambda might play in plants. The expression level of GF14 lambda under the control of the cauliflower mosaic virus 35S promoter varied in transgenic cotton plants, and lines that expressed GF14 lambda demonstrated a "stay-green" phenotype and improved water-stress tolerance. These lines wilted less and maintained higher photosynthesis than segregated non-transgenic control plants under water-deficit conditions. Stomatal conductance appears to be the major factor for the observed higher photosynthetic rates under water-deficit conditions. The stomatal aperture of transgenic plants might be regulated by GF14 lambda through some transporters such as H(+)-ATPase whose activities are controlled by their interaction with 14 3-3 proteins. However, since 14-3-3 proteins interact with numerous proteins in plant cells, many metabolic processes could be affected by the GF14 lambda overexpression. Whatever the mechanisms, the traits observed in the GF14 lambda expressing cotton plants are beneficial to crops under certain water-deficit conditions. PMID- 15356327 TI - Maintenance of growth rate at low temperature in rice and wheat cultivars with a high degree of respiratory homeostasis is associated with a high efficiency of respiratory ATP production. AB - Some plants have the ability to maintain similar respiratory rates (measured at the growth temperature) when grown at different temperatures. This phenomenon is referred to as respiratory homeostasis. Using wheat and rice cultivars with different degrees of respiratory homeostasis (H), we previously demonstrated that high-H cultivars maintained shoot and root growth at low temperature [Kurimoto et al. (2004) Plant Cell Environ., 27: 853]. Here, we assess the relationship between respiratory homeostasis and the efficiency of respiratory ATP production, by measuring the levels of alternative oxidase (AOX) and uncoupling protein (UCP), which have the potential to decrease respiratory ATP production per unit of oxygen consumed. We also measured SHAM- and CN-resistant respiration of intact roots, and the capacity of the cytochrome pathway (CP) and AOX in isolated mitochondria. Irrespective of H, SHAM-resistant respiration of intact roots and CP capacity of isolated root mitochondria were larger when plants were grown at low temperature, and the maximal activity and relative amounts of cytochrome c oxidase showed a similar trend. In contrast, CN-resistant respiration of intact roots and relative amounts of AOX protein in mitochondria isolated from those roots, were lower in high-H plants grown at low temperature. In the roots of low H cultivars, relative amounts of AOX protein were higher at low growth temperature. Relative amounts of UCP protein showed similar trends to AOX. We conclude that maintenance of growth rate in high-H plants grown at low temperature is associated with both respiratory homeostasis and a high efficiency of respiratory ATP production. PMID- 15356328 TI - Expression analysis of the NgORF13 promoter during the development of tobacco genetic tumors. AB - We investigated the expression pattern of the promoter of Nicotiana glauca (Ng) ORF13 in the hybrids between N. glauca and N. langsdorffii harboring the NgORF13 beta-glucuronidase (GUS) chimeric gene. The promoter of NgORF13 of N. glauca had lower activities than the promoter of RiORF13 of Agrobacterium rhizogenes agropine-type root-inducing (Ri) plasmid. However, the localization of GUS activity in the NgORF13 transgenic plants was similar to that in the RiORF13 transgenic plants. The GUS activity of NgORF13-GUS was high in genetic tumors cultured in vitro or developed spontaneously on F1 plants with aging or by wounding. The GUS activity in tumors was observed in bud primordia, vascular bundles and leaves in the buds. While the activity was lower than in tumors, NgORF13-GUS was also expressed in vascular bundles and the parenchymatous tissues in plants regenerated from tumors. Furthermore, the promoter activity of NgORF13 was induced by wounding and activated by exogenous application of methyl jasmonate. During tumorization, NgORF13 was induced at an early stage and showed expression patterns similar to both NgrolB and NgrolC whose expression were investigated by Nagata et al. (1996) Plant Cell Physiol. 37: 489-498. It is thought that Ngrol genes might be involved in the formation of genetic tumors, and, moreover, NgORF13 might work in cooperation with NgrolB and NgrolC. PMID- 15356329 TI - Identification of sorbitol transporters expressed in the phloem of apple source leaves. AB - Sorbitol is a major photosynthetic product and a major phloem-translocated component in Rosaceae (e.g. apple, pear, peach, and cherry). We isolated the three cDNAs, MdSOT3, MdSOT4, and MdSOT5 from apple (Malus domestica) source leaves, which are homologous to plant polyol transporters. Yeasts transformed with the MdSOTs took up sorbitol significantly. MdSOT3- and MdSOT5-dependent sorbitol uptake was strongly inhibited by xylitol and myo-inositol, but not or only weakly by mannitol and dulcitol. Apparent K(m) values of MdSOT3 and MdSOT5 for sorbitol were estimated to be 0.71 mM and 3.2 mM, respectively. The protonophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), strongly inhibited the sorbitol transport. MdSOT3 was expressed specifically in source leaves, whereas MdSOT4 and MdSOT5 were expressed in source leaves and also in some sink organs. MdSOT4 and MdSOT5 expressions were highest in flowers. Fruits showed no or only weak MdSOT expression. Although MdSOT4 and MdSOT5 were also expressed in immature leaves, MdSOT expressions increased with leaf maturation. In addition, in situ hybridization revealed that all MdSOTs were expressed to high levels in phloem of minor veins in source leaves. These results suggest that these MdSOTs are involved in sorbitol loading in Rosaceae. PMID- 15356330 TI - Cloning and functional analysis of a novel DREB1/CBF transcription factor involved in cold-responsive gene expression in Zea mays L. AB - The transcription factors DREB1s/CBFs specifically interact with the DRE/CRT cis acting element (core motif: G/ACCGAC) and control the expression of many stress inducible genes in Arabidopsis. We isolated a cDNA for a DREB1/CBF homolog, ZmDREB1A in maize using a yeast one-hybrid system. The ZmDREB1A proteins specifically bound to DRE and the highly conserved valine at the 14th residue in the ERF/AP2 DNA binding domain was a key to determining the specific interaction between this protein and the DRE sequence. Expression of ZmDREB1A was induced by cold stress and slightly increased by high-salinity stress. This gene was also transiently expressed by mechanical attack. ZmDREB1A activated the transcription of the GUS reporter gene driven by DRE in rice protoplasts. Overexpression of ZmDREB1A in transgenic Arabidopsis induced overexpression of target stress inducible genes of Arabidopsis DREB1A resulting in plants with higher tolerance to drought and freezing stresses. This indicated that ZmDREB1A has functional similarity to DREB1s/CBFs in Arabidopsis. The structure of the ERF/AP2 domain of ZmDREB1A in maize is closely related to DREB1-type ERF/AP2 domains in the monocots as compared with that in the dicots. ZmDREB1A is suggested to be potentially useful for producing transgenic plants that is tolerant to drought, high-salinity and/or cold stresses. PMID- 15356331 TI - AtIPT3 is a key determinant of nitrate-dependent cytokinin biosynthesis in Arabidopsis. AB - We analyzed the spatial expression pattern of Arabidopsis thaliana adenosine phosphates-isopentenyltransferase genes (AtIPT1, AtIPT3 to AtIPT8) and the effect of inorganic nitrogen sources on their regulation. In mature plants, the AtIPTs were differentially expressed in various tissues including the roots, leaves, stems, flowers and siliques. In transgenic seedlings expressing a gene for green fluorescent protein (GFP) driven by the AtIPT promoters, AtIPT1::GFP was predominantly expressed in the vascular stele of the roots, AtIPT3::GFP was in the phloem companion cells, AtIPT5::GFP was in the lateral root primordium and pericycle, and AtIPT7::GFP was in both the vascular stele and the phloem companion cells of the roots. In a long-term treatment, the accumulation level of AtIPT5 transcript was correlated with the concentrations of NO(3)(-) and NH(4)(+) in the growth medium. However, under nitrogen-limited conditions, AtIPT3 expression was rapidly induced by NO(3)(-) in the seedlings accompanying the accumulation of cytokinins, whereas AtIPT5 expression was little affected. The NO(3)(-)-dependent accumulation of both the AtIPT3 transcript and the cytokinins was markedly reduced in a Ds transposon-insertion mutant of AtIPT3. These results suggest that nitrogen availability differentially regulates expression of AtIPT3 and AtIPT5, and that AtIPT3 is a key determinant of cytokinin biosynthesis in response to rapid changes in the availability of NO(3)(-). PMID- 15356332 TI - Arabidopsis response regulator, ARR22, ectopic expression of which results in phenotypes similar to the wol cytokinin-receptor mutant. AB - Arabidopsis thaliana has a number of response regulators (ARRs) implicated in the histidine (His)-->aspartate (Asp) phosphorelay signal transduction. According to the current consistent model, both the type-A and type-B ARR family members play crucial roles in the cytokinin signaling circuitry. However, this higher plant has a few extra ARRs, on which no attention has been paid so far. Characterization of these extra ARRs might provide us with new insight into the His-->Asp phosphorelay signal transduction in plants. For this reason, in this study we extensively examined the natures of such a representative (named ARR22). Transcripts of ARR22 were expressed predominantly in reproductive organs, and a GFP::ARR22 fusion protein was localized in the cytoplasmic space in onion epidermal cells. The purified ARR22 protein had the ability to undergo phosphorylation in vitro, when incubated with phospho-AHP5, indicating that ARR22 has the fundamental ability to participate into a His-Asp phosphorelay pathway in its own right. In plants, transgenic lines overexpressing ARR22 were characterized (referred to as ARR22-ox), which showed the characteristic dwarf phenotypes with poorly developed root systems. The results of Northern blot hybridization with selected sets of hormone-responsive genes suggested that cytokinin responses are selectively attenuated in ARR22-ox, while other hormone responses (auxin, ABA and ethylene) occur normally. The results of microarray analyses with cytokinin-treated wild-type and ARR22-ox plants further supported the view that cytokinin responses are globally attenuated in ARR22-ox, at least, at the level of gene regulation. Finally, we demonstrated that the dwarf phenotypes of ARR22-ox are very similar to those of the wooden leg (wol) mutant, which has a severe lesion in the AHK4/CRE1 cytokinin-receptor of histidine protein kinase. These results suggested that ARR22 might also be implicated, directly or indirectly, in the cytokinin-responsive His-->Asp phophorelay signal transduction. PMID- 15356333 TI - Circadian-controlled basic/helix-loop-helix factor, PIL6, implicated in light signal transduction in Arabidopsis thaliana. AB - PHYTOCHROME-INTERACTING FACTOR-LIKE 6 (PIL6) is a member of the large family of basic/helix-loop-helix (bHLH) factors in Arabidopsis thaliana. This circadian controlled transcription factor was previously suggested to interact with the clock component, TIMING OF CAB EXPRESSION 1 (TOC1). In this study, we isolated a loss-of-function mutant of PIL6, together with a transgenic line aberrantly expressing PIL6 in a manner independent of circadian rhythm. These mutant plants were simultaneously examined with special reference to circadian rhythm and light signal transduction. The results suggested that PIL6 appears to be not directly involved in the clock function per se. However, the loss-of-function mutant (pil6 1) showed a remarkable phenotype in that it is hypersensitive to red light in seedling de-etiolation. This phenotype was similar to that observed for transgenic lines overexpressing TOC1 (or APRR1). Conversely, transgenic plants overexpressing PIL6 (PIL6-ox) are hyposensitive to red light under the same conditions. This phenotype was very similar to that observed for phyB mutants. The developmental morphologies of PIL6-ox, including the phenotype of early flowering, were also similar to those of phyB mutants. We propose that PIL6 acts as a negative regulator for a red light-mediated morphogenic response (e.g., elongation of hypocotyls in de-etiolation). Taken together, PIL6 might function at an interface between the circadian clock and red light-signal transduction pathways. PMID- 15356334 TI - Localization of glutamine synthetase isoforms in hair cells of Azolla leaves. AB - Immunoelectron microscopy and a quantitative analysis of immunogold labeling of a glutamine synthetase (GS; EC 6.3.1.2) revealed that, in mesophyll cells of mature leaves of Azolla filiculoides, almost all GS was present in chloroplasts. By contrast, in hair cells, abundant labeling of GS was observed both in chloroplasts and in the cytoplasm. In hair cells of cyanobiont-free plants, the labeling of GS of both chloroplasts and cytoplasm was very weak compared to that of cyanobiont-containing plants. The findings suggest that hair cells play an important role in the assimilation of ammonia released by the cyanobiont. PMID- 15356335 TI - A recombinant plant natriuretic peptide causes rapid and spatially differentiated K+, Na+ and H+ flux changes in Arabidopsis thaliana roots. AB - Plant natriuretic peptides (PNPs) belong to a novel class of systemically mobile molecules that are structurally similar to the N-terminal domain of expansins and affect physiological processes such as protoplast volume regulation at nano-molar concentrations. Here we demonstrate that AtPNP-A, a recombinant Arabidopsis thaliana PNP causes rapid H(+) influx in the elongation zone of A. thaliana roots but not in the mature zone. AtPNP-A also induces significant K(+) and Na(+) efflux and this effect is seen in the mature root zone only. These observations suggest that responses to AtPNP-A are developmental stage and tissue specific and point to a complex role in plant growth and homeostasis. PMID- 15356336 TI - Stable isotope labeling of Arabidopsis thaliana for an NMR-based metabolomics approach. AB - Nuclear magnetic resonance (NMR) will become a key technology in plant metabolomics with the use of stable isotope labeling and advanced hetero-nuclear NMR methodologies. To demonstrate the power of this approach, we performed multi dimensional hetero-nuclear NMR analysis of metabolic movement of carbon and nitrogen nuclei in Arabidopsis thaliana. First, distinct ethanol-stress response was investigated using (13)C-labeled wild type and an ethanol-hypersensitive mutant plants. Furthermore, we followed nitrogen fluxes in (15)N-labeled seeds during the initiation of germination in vivo. The future role of stable isotope labeling combined with advanced hetero-nuclear NMR in plant metabolomics is discussed. PMID- 15356337 TI - A rice antisense SPK transformant that lacks the accumulation of seed storage substances shows no correlation between sucrose concentration in phloem sap and demand for carbon sources in the sink organs. AB - Rice SPK is a calmodulin-like domain protein kinase specific to immature seeds and promotes the degradation of sucrose. Therefore, antisense SPK transformants showed a defective production of storage starch, but accumulated sucrose in watery seeds. Despite a reduced sink strength, no difference was found in the sucrose concentration in phloem sap of the transformants and wild-type plants, which increased after floral organ induction to levels greater than 500 mM. However, sucrose was detected at relatively lower levels in the watery seed sap. These results suggest that sucrose content in the phloem is regulated independently from the demand for carbon sources in the sink organs. PMID- 15356338 TI - Marek's disease is a natural model for lymphomas overexpressing Hodgkin's disease antigen (CD30). AB - Animal models are essential for elucidating the molecular mechanisms of carcinogenesis. Hodgkin's and many diverse non-Hodgkin's lymphomas overexpress the Hodgkin's disease antigen CD30 (CD30(hi)), a tumor necrosis factor receptor II family member. Here we show that chicken Marek's disease (MD) lymphoma cells are also CD30(hi) and are a unique natural model for CD30(hi) lymphoma. Chicken CD30 resembles an ancestral form, and we identify a previously undescribed potential cytoplasmic signaling domain conserved in chicken, human, and mouse CD30. Our phylogeneic analysis defines a relationship between the structures of human and mouse CD30 and confirms that mouse CD30 represents the ancestral mammalian gene structure. CD30 expression by MD virus (MDV)-transformed lymphocytes correlates with expression of the MDV Meq putative oncogene (a c-Jun homologue) in vivo. The chicken CD30 promoter has 15 predicted high-stringency Meq-binding transcription factor recognition motifs, and Meq enhances transcription from the CD30 promoter in vitro. Plasma proteomics identified a soluble form of CD30. CD30 overexpression is evolutionarily conserved and defines one class of neoplastic transformation events, regardless of etiology. We propose that CD30 is a component of a critical intracellular signaling pathway perturbed in neoplastic transformation. Specific anti-CD30 Igs occurred after infection of genetically MD-resistant chickens with oncogenic MDV, suggesting immunity to CD30 could play a role in MD lymphoma regression. PMID- 15356339 TI - Kinase peptide specificity: improved determination and relevance to protein phosphorylation. AB - Specificity of phosphorylation is critical to signal transduction. Recent emphasis on colocalization of substrate and kinase has eclipsed emphasis on peptide specificity, i.e., kinase preference for particular amino acids surrounding the phosphorylation site. We describe an approach to determining peptide specificity by using positional scanning of biotinylated oriented peptide libraries and insights emerging from those determinations. We accurately determine preference (or disfavor) for residues at a given substrate position (such as P+2) by comparison of in vitro phosphorylation of peptide libraries differing by a single residue at that position. By analysis of all positions near the phosphorylation site, position-specific scoring matrices are generated and used both to understand the basis of specificity and to predict phosphorylation. PKC-delta and -zeta predictions have been validated rigorously by comparisons with measured phosphorylation. The results demonstrate specificity and sensitivity (80-90%) much better than the previous predictive method. These predictions can be accessed at http://mpr.nci.nih.gov. The accuracy of the specificity determination allows identification of an important difference in peptide specificity between these closely related kinases; Ile/Leu at the P-1 position is disfavored by PKC-zeta but not PKC-delta. Our findings and visual representation of peptide specificity highlight the importance of disfavored residues. Finally, analysis of 124 experimentally determined PKC sites from the literature demonstrates a very strong role of peptide specificity in many of those sites. Thus, position-specific scoring matrices generated by this method provide a foundation for quantitative analyses of kinase specificity and improved predictions of previously determined physiologically relevant phosphorylation sites. PMID- 15356340 TI - The influence of sulfur and iron on dissolved arsenic concentrations in the shallow subsurface under changing redox conditions. AB - The chemical speciation of arsenic in sediments and porewaters of aquifers is the critical factor that determines whether dissolved arsenic accumulates to potentially toxic levels. Sequestration of arsenic in solid phases, which may occur by adsorption or precipitation processes, controls dissolved concentrations. We present synchrotron x-ray absorption spectra of arsenic in shallow aquifer sediments that indicate the local structure of realgar (AsS) as the primary arsenic-bearing phase in sulfate-reducing conditions at concentrations of 1-3 mmol.kg(-1), which has not previously been verified in sediments at low temperature. Spectroscopic evidence shows that arsenic does not substitute for iron or sulfur in iron sulfide minerals at the molecular scale. A general geochemical model derived from our field and spectroscopic observations show that the ratio of reactive iron to sulfur in the system controls the distribution of solid phases capable of removing arsenic from solution when conditions change from oxidized to reduced, the rate of which is influenced by microbial processes. Because of the difference in solubility of iron versus arsenic sulfides, precipitation of iron sulfide may remove sulfide from solution but not arsenic if precipitation rates are fast. The lack of incorporation of arsenic into iron sulfides may result in the accumulation of dissolved As(III) if adsorption is weak or inhibited. Aquifers particularly at risk for such geochemical conditions are those in which oxidized and reduced waters mix, and where the amount of sulfate available for microbial reduction is limited. PMID- 15356341 TI - Learning increases human electroencephalographic coherence during subsequent slow sleep oscillations. AB - Learning is assumed to induce specific changes in neuronal activity during sleep that serve the consolidation of newly acquired memories. To specify such changes, we measured electroencephalographic (EEG) coherence during performance on a declarative learning task (word pair associations) and subsequent sleep. Compared with a nonlearning control condition, learning performance was accompanied with a strong increase in coherence in several EEG frequency bands. During subsequent non-rapid eye movement sleep, coherence only marginally increased in a global analysis of EEG recordings. However, a striking and robust increase in learning dependent coherence was found when analyses were performed time-locked to the occurrence of slow oscillations (<1 Hz). Specifically, the surface-positive half waves of the slow oscillation resulting from widespread cortical depolarization were associated with distinctly enhanced coherence after learning in the slow oscillatory, delta, slow-spindle, and gamma bands. The findings identify the depolarizing phase of the slow oscillations in humans as a time period particularly relevant for a reprocessing of memories in sleep. PMID- 15356342 TI - Visualization of plasma membrane compartmentalization with patterned lipid bilayers. AB - Micrometer-size patterned lipid bilayers containing liganded lipids are used to control the location and size of receptor clusters and enable direct visualization of structural reorganization of cellular components. Subsequent to concentration of Fcepsilon receptor I, the mast cell receptor for IgE, and colocalized tyrosine phosphorylation activity, Lyn kinase and other proteins anchored to the inner leaflet of the plasma membrane redistribute selectively with the receptor clusters in a process that depends on actin polymerization. Surprisingly, outer leaflet components characteristically associated with lipid rafts do not detectably coredistribute with these inner leaflet components. Cell activation using patterned surfaces provides unique insights into cell membrane structural organization, revealing dynamic, large-scale uncoupling of inner and outer leaflet components of lipid rafts. PMID- 15356343 TI - Carcinoma and stromal enzyme activity profiles associated with breast tumor growth in vivo. AB - Cancer research depends on the use of human cell lines for both the in vitro (culture) and in vivo (xenograft) analysis of tumor progression and treatment. However, the extent to which cultured preparations of human cancer lines display similar properties in vivo, where important host factors may influence tumor biology, remains unclear. Here, we address this question by conducting a functional proteomic analysis of the human breast cancer line MDA-MB-231 grown in culture and as orthotopic xenograft tumors in the mammary fad pad of immunodeficient mice. Using a suite of activity-based chemical probes, we identified carcinoma (human) enzyme activities that were expressed selectively in culture or in xenograft tumors. Likewise, distinct groups of stromal (mouse) enzyme activities were found that either infiltrated or were excluded from xenograft tumors, indicating a contribution by specific host components to breast cancer development. MDA-MB-231 cells isolated from tumors exhibited profound differences in their enzyme activity profiles compared with the parent cell line, including the dramatic posttranscriptional up-regulation of the serine proteases urokinase plasminogen activator and tissue plasminogen activator and down regulation of the glycolytic enzyme phosphofructokinase. These altered enzyme activity profiles correlated with significantly greater tumor growth rates and metastases for xenograft-derived MDA-MB-231 cells upon reintroduction into mice. Collectively, these data indicate that the in vivo environment of the mouse mammary fat pad cultivates the growth of human breast cancer cells with elevated tumorigenic properties and highlight the value of activity-based protein profiling for identifying proteomic signatures that depict such changes in cancer cell biology. PMID- 15356344 TI - Structural analysis uncovers a role for redox in regulating FKBP13, an immunophilin of the chloroplast thylakoid lumen. AB - Change in redox status has long been known to link light to the posttranslational regulation of chloroplast enzymes. So far, studies have been conducted primarily with thioredoxin-linked members of the stroma that function in a broad array of biosynthetic and degradatory processes. Consequently, little is known about the role of redox in regulating the growing number of enzymes found to occur in the lumen, the site of oxygen evolution in thylakoid membranes. To help fill this gap, we have studied AtFKBP13, an FKBP-type immunophilin earlier shown to interact with a redox-active protein of the lumen, and found the enzyme to contain a pair of disulfide bonds in x-ray structural studies. These disulfides, which in protein mutagenesis experiments were shown to be essential for the associated peptidyl-prolyl isomerase activity, are unique to chloroplast FKBPs and are absent in animal and yeast counterparts. Both disulfide bonds were redox active and were reduced by thioredoxin from either chloroplast or bacterial sources in a reaction that led to loss of enzyme activity. The results suggest a previously unrecognized paradigm for redox regulation in chloroplasts in which activation by light is achieved in concert with oxygen evolution by the oxidation of sulfhydryl groups (conversion of SH to S-S). Such a mechanism, occurring in the thylakoid lumen, is in direct contrast to regulation of enzymes in the stroma, where reduction of disulfides targeted by thioredoxin (S-S converted to SH) leads to an increase in activity in the light. PMID- 15356346 TI - Increased susceptibility to adult paraoxon exposure in mice neonatally exposed to nicotine. AB - Low-dose exposure of neonatal mice to nicotine has earlier been shown to induce an altered behavioral response to nicotine in adulthood. Organophosphorus insecticides are known to affect the cholinergic system by inhibition of acetylcholinesterase. This study was undertaken to investigate whether neonatal exposure to nicotine makes mice more susceptible to a known cholinergic agent. Neonatal, 10-day-old, male mice were exposed to nicotine-base (33 microg/kg body weight) or saline s.c. twice daily on five consecutive days. At 5 months of age the animals were exposed to paraoxon (0.17 or 0.25 mg/kg body weight [29% and 37% inhibition of cholinesterase, respectively]) or saline sc every second day for 7 days. Before the first paraoxon injection, the animals were observed for spontaneous motor behavior. The spontaneous motor behavior test did not reveal any differences in behavior between the treatment groups. Immediately after the spontaneous behavior test, the animals received the first injection of paraoxon and were observed for acute effects of paraoxon on spontaneous motor behavior. The acute response to paraoxon in the spontaneous motor behavior test was a decreased level of activity in mice neonatally exposed to nicotine. Control animals showed no change in activity. Two months after the paraoxon treatment, the animals were again tested for spontaneous motor behavior. Animals neonatally exposed to nicotine and exposed to paraoxon as adults showed a deranged spontaneous motor behavior, including hyperactivity and lack of habituation. PMID- 15356345 TI - Protein tyrosine phosphatase receptor-type O (PTPRO) exhibits characteristics of a candidate tumor suppressor in human lung cancer. AB - Previous study in our laboratory demonstrated suppression of the gene for protein tyrosine phosphatase receptor-type O (PTPRO) in primary and established rat hepatomas. The present study showed methylation-mediated silencing of this gene in primary human lung tumors and in several human lung cancer cell lines, one of the characteristics of many tumor-suppressor genes. The reduced expression of PTPRO in the primary lung tumors correlated with the methylation status of its CpG island. Demethylation of the gene by deoxy-5-azacytidine treatment led to its reactivation in a lung cancer line (A549). Overexpression of PTPRO in A549 cells inhibited anchorage-independent growth, delayed reentry of the cells into the cell cycle after release from cell-cycle arrest, and increased susceptibility of the cells to apoptosis. These data have demonstrated the growth-suppressor characteristics of PTPRO that are unique to a classical tumor suppressor. PMID- 15356347 TI - Leashing Leishmania. Some old mice fight parasitic infection better than young mice do. PMID- 15356348 TI - Will a filet a day keep the neurologist away?Fish oil molecule forestalls neuron damage in Alzheimer's mice. PMID- 15356349 TI - TOR pathway: linking nutrient sensing to life span. AB - Recent studies point to an emerging role for the TOR (target of rapamycin) pathway in the regulation of life span. In this Perspective, we discuss the possibility that this pathway is an important modulator of nutrient-dependent changes in life span. Additionally, we discuss the interactions between the TOR and insulin-like signaling pathways as well as the key downstream processes that TOR regulates. PMID- 15356351 TI - Endocrine emergencies. AB - Diabetic and endocrine emergencies are traditionally treated by the acute medical admitting team or accident and emergency department staff. Most will see diabetic emergencies on a regular basis, as they are common and both type 1 and type 2 disease are increasing in prevalence. Diabetic emergencies are usually easily treated and the patients discharged. However, it is vital not to become complacent as these disorders can lead to death. It is particularly important to follow local guidance and to involve the diabetes team both during and after each episode. Recently it has become clear that about 30% of patients admitted with acute coronary syndrome (including infarction) have either diabetes or "stress hyperglycaemia"; evidence suggests that these patients should be treated not only as a cardiac emergency but also as a diabetic one. Thus, every patient with acute coronary syndrome or acute myocardial infarction needs diabetes to be excluded. The other endocrine emergencies are less common, but in some ways more important simply because of their rarity. A high level of suspicion is often required to make a diagnosis, although some, such as myxoedema coma, are usually obvious. Treatment must be started before the diagnosis can be confirmed. Guidance on making the diagnosis and initiating treatment should be made available on the local NHS intranet for non-endocrinologists to access; and where possible expert advice made available by telephone. The basic management steps in the common diabetic and endocrine emergencies are outlined; this is not a complete list, but rather an insight for those involved in non-selected emergency admissions. PMID- 15356350 TI - Chronic obstructive pulmonary disease: the clinical management of an acute exacerbation. AB - Exacerbations of chronic obstructive pulmonary disease impose a considerable burden of morbidity, mortality, and health care cost. Management guidelines outlining best practice, based largely on consensus expert opinion, were produced by a number of organisations during the last decade. Current interest in the field is high. This has resulted in the publication of many further studies which have extended our understanding of the pathology involved and provided, for the first time, an evidence base for many of the therapeutic options. In this review we aim to bring the non-specialist reader up to date with current management principles and the evidence underlying such interventions. PMID- 15356353 TI - Cholecystocolic fistula demonstrated by endoscopic retrograde cholangiopancreatography. PMID- 15356352 TI - Probiotics and human health: a clinical perspective. AB - There is unequivocal evidence that administration of probiotics could be effective in the treatment of acute infectious diarrhoea in children and the prevention of antibiotic associated diarrhoea and nosocomial/community acquired diarrhoea. Encouraging evidence is also emerging for the effectiveness of probiotics in the prevention and management of pouchitis and paediatric atopic diseases, and the prevention of postoperative infections. There is also strong evidence that certain probiotic strains are able to enhance immune function, especially in subjects with less than adequate immune function such as the elderly. Efficacy of probiotics in the prevention of traveller's diarrhoea, sepsis associated with severe acute pancreatitis, and cancers, the management of ulcerative colitis, and lowering of blood cholesterol remains unproven. In addition to firm evidence of efficacy (for a range of conditions), major gaps exist in our knowledge regarding the mechanisms by which probiotics modulate various physiological functions and the optimum dose, frequency, and duration of treatment for different probiotic strains. PMID- 15356354 TI - Chorea and related disorders. AB - Chorea refers to irregular, flowing, non-stereotyped, random, involuntary movements that often possess a writhing quality referred to as choreoathetosis. When mild, chorea can be difficult to differentiate from restlessness. When chorea is proximal and of large amplitude, it is called ballism. Chorea is usually worsened by anxiety and stress and subsides during sleep. Most patients attempt to disguise chorea by incorporating it into a purposeful activity. Whereas ballism is most often encountered as hemiballism due to contralateral structural lesions of the subthalamic nucleus and/or its afferent or efferent projections, chorea may be the expression of a wide range of disorders, including metabolic, infectious, inflammatory, vascular, and neurodegenerative, as well as drug induced syndromes. In clinical practice, Sydenham's chorea is the most common form of childhood chorea, whereas Huntington's disease and drug induced chorea account for the majority of adult onset cases. The aim of this review is to provide an up to date discussion of this disorder, as well as a practical approach to its management. PMID- 15356355 TI - Management of paediatric asthma. AB - Paediatric asthma best practice not only includes prescribing the correct therapeutic mix based on consensus guidelines, but also reducing therapy once control has been achieved. Clinicians should also be aware that asthma in young children is a heterogeneous entity, and a beneficial response to bronchodilators and/or inhaled steroids is not inevitable. In general, preschool children and infants should not be prescribed inhaled corticosteroids above 200 microg beclometasone dipropionate equivalent twice a day, or regular oral steroids, or long acting beta2-adrenoceptor agonists. New therapies such as anti-IgE antibodies are on the horizon, but these are unlikely to replace the established drug combinations. More likely is that the delivery of established drugs will become more convenient (for example, once a day inhaled corticosteroids, or season dependent prophylactic therapy). PMID- 15356356 TI - Salmonella: a continuing problem. AB - Salmonella was the archetypal food poisoning organism of the 20th century. It achieved a high public profile following the salmonella-in-eggs crisis of 1988 89, but by then had been the subject of public health concern and scientific interest for over a century. Early associated with animal foods, the advent of phage typing in the 1940s began to reveal the complexity of its environmental associations. This paper explores the story of salmonella as a continuing problem in epidemiology, microbiology and public health, in the food chain, and in the kitchen. PMID- 15356358 TI - The embryonic explanation for dermatome distributions. PMID- 15356357 TI - Sentinel lymph node biopsy in breast cancer patients after overnight migration of radiolabelled sulphur colloid. AB - OBJECTIVE: To evaluate the performance and feasibility of sentinel lymph node biopsy in breast cancer patients using technetium-99m (99mTc) sulphur colloid and gamma probe. METHODS: From May 2000 to March 2001, 70 patients with a tumour less than 5 cm with clinically negative axillary lymph nodes underwent sentinel node biopsy followed by standard axillary dissection. 99mTc sulphur colloid was injected around the primary tumour the day before surgery and a gamma probe was used to detect the sentinel lymph node during the surgical procedure. Sentinel lymph node biopsy was compared with standard axillary dissection for its ability to accurately reflect the final pathological status of the axillary nodes. RESULTS: The sentinel lymph node was successfully identified in 67 of 70 patients (95.71%). The number of sentinel lymph nodes ranged from 1-5 (mean 1.5) and non sentinel nodes ranged from 5-22 (mean 13.3). Of the 67 patients with successfully identified sentinel lymph nodes, 43.28% (29/67) were histologically positive. Sensitivity of the sentinel lymph node to predict axilla was 82.75%; specificity was 100%. Positive and negative predictive values were 100% and 88.3% respectively. The sentinel lymph node was falsely negative in five patients, yielding an accuracy of 92.53%. Sentinel lymph node biopsy was more accurate for T1 tumours than for T2 tumours. CONCLUSIONS: The gamma probe guided method after overnight migration of 99mTc sulphur colloid is technically feasible for detecting sentinel lymph nodes in most breast cancer patients, accurately predicting the axillary lymph node status, and appears more accurate for T1 lesions than for larger lesions. This minimally invasive axillary staging procedure represents a major advance in the surgical treatment of breast cancer. PMID- 15356359 TI - Thrombotic thrombocytopenic purpura associated with statin treatment. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare condition but associated with 90% mortality if left untreated. The diagnosis is usually made when there is thrombocytopenia and microangiopathic haemolytic anaemia, although the full pentad also includes fever, renal impairment, and neurological dysfunction. A variety of underlying causes have been implicated in acquired TTP including bacterial and viral infections, bone marrow and organ transplantation, pregnancy, immune disorders, and certain drugs. To date there is just one case report of TTP associated with statin treatment. The clinical course of a patient who presented with TTP after being started on simvastatin, a HMG-CoA inhibitor, is described. PMID- 15356360 TI - Fulminant liver failure: an indicator of silent myocardial rupture. AB - A 56 year old man presented with an atypical chest infection. Remote inferoposterior myocardial infarction was noted on electrocardiography and transthoracic echocardiography. Hepatic failure developed with sudden gross elevation of liver aminotransferases and coagulopathy. No primary hepatic cause could be identified. Subsequent right heart failure led to transoesophageal echocardiography that revealed a large inoperable ventricular septal defect. Histopathological data showed ischaemic hepatitis and reinfarction of the inferoposterior myocardial wall. Acute cardiac events may be silent and precipitate misleading severe hepatic dysfunction. PMID- 15356361 TI - A man with drowsiness and abdominal pain. PMID- 15356362 TI - Painful knee. PMID- 15356363 TI - The NPUAP Support Surface Standards Initiative. PMID- 15356364 TI - Pain management in epidermolysis bullosa: an intractable problem. PMID- 15356365 TI - The king's royal urine. PMID- 15356366 TI - MMA provisions begin. PMID- 15356367 TI - A comparison of an antimicrobial wound cleanser to normal saline in reduction of bioburden and its effect on wound healing. AB - Microbial bioburden in both acute and chronic wounds is an important factor in wound healing. Consequently, the reduction of bioburden to host-manageable levels, as well as the elimination of certain virulent forms of wound pathogens (regardless of their number), has become a goal of the wound care professional. A prospective, controlled clinical study using accepted sampling methods was conducted to compare the use of an antimicrobial wound cleanser (0.057% sodium hypochlorite in an isotonic saline solution) to normal saline on the reduction of bioburden and wound size. During the 2-month study, 100% of the wounds cleansed with the antimicrobial wound cleanser (n = 9) demonstrated aerobic bioburden reduction from baseline in a range from 1 to 4 logs per wound, while 56% of the wounds cleansed with normal saline (n = 9) showed an increase in aerobic bioburden levels. The proportion of wounds exhibiting a reduction in wound size was higher in the antimicrobial wound cleanser group than in the saline group. Further research to increase understanding of the relationship between wound bioburden, healing, and cleansing agents is needed. PMID- 15356368 TI - Impact of antimicrobial gauze on bacterial colonies in wounds that require packing. AB - Heavy microbial contamination has been associated with delayed wound healing and infection in both acute and chronic wounds. A prospective, randomized, 5-week controlled, open label, multicenter study was conducted to determine whether using antimicrobial gauze containing polyhexamethylene biguanide in wounds that require packing will result in a greater reduction of bacterial colony counts than using a gauze without polyhexamethylene biguanide (the control). Twenty-one subjects were randomized to the treatment or control dressing. Wounds were evenly distributed with respect to etiology and both study groups had a median baseline wound size of 7 cm2. At baseline, 15 microbial isolates were recovered and counted in treatment group wounds and 12 were recovered in the controls. At Week 1, six isolates were recovered from subjects in the polyhexamethylene biguanide antimicrobial gauze treatment group while 10 were recovered in the control. Change in polymicrobial bioburden was most prominent during the first few weeks of the study in the polyhexamethylene biguanide group. Polymicrobial counts in the treatment group remained reduced for the following three study weeks, returning to baseline at Week 4. In the control group, the number of polymicrobial cultures rose to 60% above baseline at Week 4. Two wounds of subjects randomized to the polyhexamethylene biguanide antimicrobial gauze healed; one wound in the control group healed. Polyhexamethylene biguanide antimicrobial gauze dressing could be an important adjunct to control the polymicrobial bioburden of delayed closure surgical wounds, pressure ulcers, and diabetic foot ulcers. Additional studies seem warranted. PMID- 15356369 TI - Bedrails: restraints or enablers? AB - Bedrails presently are used as both mobility restraints and enablers in long-term care facilities. As enablers, bedrails facilitate movement and may reduce the risk of pressure ulcer development. As restraints, they impede movement and may increase risk of ulcer development. Omnibus Budget Reconciliation Act regulations on restraint use have led to confusion for state Medicare surveyors and facilities regarding the definition of appropriate bedrail use and need for supportive documentation. Consequently, some facilities receive deficiency citations for inappropriate use or documentation while others do not. The purpose of this survey was to compare responses of Directors of Nursing in long-term care facilities and Medicare state surveyors to determine how each interprets the Omnibus Budget Reconciliation Act bedrail language for use and documentation. Questionnaires on bedrail use and documentation were sent to state surveyors and Directors of Nursing. One hundred, three (103) Directors of Nursing in 45 states and 65 surveyors from 39 states participated in the survey (response rate 61%). Study results demonstrated general acceptance of bedrail use as an enabler but not as a restraint by both Directors of Nursing and state surveyors. Four percent (4%) of Directors of Nursing reported receiving a citation for bedrail use and 59% of surveyors reported issuing citations for bedrail use. Significant differences were noted between the two groups regarding appropriate bedrail use and necessary documentation. The intent of Medicare guidelines and the Centers for Medicare and Medicaid Services is to standardize care for nursing home residents in the United States; yet, current regulations are open to individual interpretation by state surveyors and confusion exists between the intent of the Omnibus Budget Reconciliation Act and the daily operations of nursing homes. Educating clinicians about the risks and benefits of bedrail use, either as restraint or enabler, and developing interventions and policies for appropriate use would be an important first step in resolving this issue. PMID- 15356370 TI - "Venerate the Lancet": Benjamin Rush's yellow fever therapy in context. AB - In 1793, during a yellow fever epidemic in Philadelphia, Benjamin Rush adopted a therapy that centered on rapid depletion through purgation and bleeding. His method, especially his reliance on copious bloodletting, was at first widely condemned, but many American practitioners eventually adopted it. Although the therapy struck many observers as being radical, in large part it grew from premises that had substantial support. Rush was convinced that it worked and that heroic methods were the key to conquering disease. In particular, massive bleeding became central to his therapeutics. PMID- 15356371 TI - Averting disaster: the Hudson's Bay Company and smallpox in western Canada during the late eighteenth and early nineteenth centuries. AB - During the late eighteenth and early nineteenth centuries the Hudson's Bay Company served as a de facto public health agency across western Canada. Among its biggest challenges was combating the smallpox epidemics that periodically threatened the Aboriginal people of the region. Initially, the Company's employees turned to quarantine over variolation in order to prevent the spread of the disease to Hudson Bay in the summer of 1782. Although well thought-out, ultimately this policy proved unsuccessful. Within thirty years the HBC had turned to the newly discovered vaccination, a strategy that was to prove far more effective in fighting the disease. By the late 1830s the Company was able to mount an effective vaccination campaign that covered much of western Canada. PMID- 15356372 TI - A duty to kill? A duty to die? Rethinking the euthanasia controversy of 1906. AB - This essay seeks to chronicle the controversy surrounding euthanasia that came to a head in the United States around 1906, and to situate that debate in a meaningful historical context. An extensive examination of newspaper coverage of the legislative proposals to legalize the practice in Ohio and Iowa reveals that (1) the discourse occurred in a context in which both supporters and opponents of euthanasia generally agreed that the practice already occurred with frequency; (2) the discussion was heavily influenced by a simultaneous controversy surrounding eugenics; and (3) most of the opponents of legalization relied upon practical rather than religious or moral arguments to further their cause. PMID- 15356373 TI - Patterns of municipal health expenditure in interwar England and Wales. AB - This article aims to fill a gap in the history of medical services in England and Wales in the interwar period by focusing on the historiographically neglected municipal sector--a relative neglect that is particularly unjustified given that this sector provided an increasingly wide array of medical services over the period. Focusing on the highly urbanized county boroughs, this article investigates whether and how expenditure on municipal health services changed over the interwar period, and whether these patterns were replicated by boroughs across England and Wales. It is found that many of the largest personal health services were experiencing a common pattern of growing investment over the period, but that county boroughs did not act uniformly in their spending decisions. Considered regionally, the Northeast and the West Midlands were found to perform poorly in expenditure terms compared to the data set as a whole, while the large conurbations of Leeds, Manchester, and Liverpool raised the average performance of the Northwest and Yorkshire. Regional patterns are found to be less consistent in the south of the country, where voluntary provision and demands arising from the boroughs' geographical position (for example, seaside resorts) may have exerted significant influences over levels of expenditure on health. PMID- 15356377 TI - Correlation of vulnerable coronary plaques to sudden cardiac events. Lessons from a myocardial infarction-prone animal model (the WHHLMI rabbit). AB - It is generally considered that coronary rupture-prone plaques play an important role in the onset of sudden cardiac events (acute coronary syndromes/sudden cardiac death). However, it is not clear which factors or stimuli are required to trigger plaque rupture and whether coronary plaques without occlusive thrombi can cause sudden cardiac events. To address these issues, recently, we developed a rabbit model of spontaneous myocardial infarction [the Watanabe heritable hyperlipidemic (WHHL) MI rabbit] and found that this model possessed several types of coronary plaques that are possibly correlated to sudden cardiac events. Although many of the coronary plaques of the WHHLMI rabbits appeared histologically to be rupture-prone in nature, true rupture was detected only in the few animals that died of MI. In addition, no occlusive thrombus was detected in any WHHLMI rabbit. These findings suggest that some additional stimuli play a definitive role in causing disruption of rupture-prone plaques and thrombosis. Nearly-occluded plaques caused by a luminal macrophage accumulation are the most common feature of WHHLMI rabbits, suggesting that they are responsible for sudden cardiac events. The WHHLMI rabbit could be a useful model for studying the mechanism(s) of plaque rupture and thrombogenesis if plaque rupture/thrombus formation is induced in the rupture-prone plaques of WHHLMI rabbits by administration of additional triggering factors, and could provide a novel means for developing new therapies. PMID- 15356378 TI - Lipid analyses for the management of vascular diseases. AB - Despite a long history of studies on lipid abnormalities, numerous problems in laboratory technologies and techniques remain unresolved. One of the most commonly tested molecules is low-density lipoprotein (LDL) cholesterol, and a homogenous assay technique for measurement of this molecule has recently been introduced. Although the method represents remarkable technological breakthroughs with great potential for improving lipoprotein analysis, some discrepancies exist among assay protocols. Even for direct measurement of high-density lipoprotein (HDL) cholesterol, which has widely been accepted, there are still large discrepancies among data obtained by different protocols. Oxidatively modified LDL is an independent factor that is considered to directly contribute to the pathogenesis of early atherosclerosis. Lipid peroxidation products, surface charge, and spectrophotometric patterns are all applicable to the evaluation of in vitro oxidation. Only enzyme-linked immunosorbent assays using monoclonal antibodies have a potential for clinical use, but such methods are not yet standardized. There is also increasing evidence for the presence of anti-oxidized LDL autoantibodies in human sera, but the diagnostic utility remains controversial. In addition, small dense LDL has recently attracted much attention as an independent risk factor. Although this is a potential target of oxidization, a robust and simple analytical method does not yet exist. This review presents the current state of laboratory technologies for testing lipid abnormalities. PMID- 15356379 TI - The very low-density lipoprotein (VLDL) receptor: characterization and functions as a peripheral lipoprotein receptor. AB - The very low-density lipoprotein (VLDL) receptor is a member of the low-density lipoprotein (LDL) receptor family. In vitro and in vivo studies have shown that VLDL receptor binds triglyceride (TG)-rich lipoproteins but not LDL, and functions as a peripheral remnant lipoprotein receptor. VLDL receptor is expressed abundantly in fatty acid-active tissues (heart, skeletal muscle and fat), the brain and macrophages. It is likely that VLDL receptor functions in concert with lipoprotein lipase (LPL), which hydrolyses TG in VLDL and chylomicron. In contrast to the LDL receptor, VLDL receptor binds apolipoprotein (apo) E2/2 VLDL particles as well as apoE3/3 VLDL, and the expression is not down regulated by intracellular lipoproteins. Recently, various functions of the VLDL receptor have been reported in lipoprotein metabolism, metabolic syndrome/atherosclerosis, cardiac fatty acid metabolism, neuronal migration and angiogenesis/tumor growth. Gene therapy of VLDL receptor into the liver showed a benefit effect for lipoprotein metabolism in both LDL receptor knockout and apoE mutant mice. Beyond its function as a peripheral lipoprotein receptor, possibilities of its physiological function have been extended to include signal transduction, angiogenesis and tumor growth. PMID- 15356380 TI - Reduction of visceral adiposity after operation in a subject with insulinoma. AB - Generally, it is considered that visceral fat brings insulin resistance and hyper insulinemia, in the mechanisms of metabolic syndromes. However, whether hyperinsulinemia brings about accumulation of visceral fat is not clear. We followed a case of insulinoma that caused primary hyperinsulinemia, and measured the change in visceral fat and insulin resistance before and after surgical resection of the insulinoma. A 58-year-old woman was admitted to investigate the cause of spontaneous hypoglycemia. An oral glucose tolerance test (OGTT) showed hyperinsulinemia with a high basal level and a glucagon infusion test showed an abnormally high insulin level. Abdominal computed tomography (CT) scan showed an accumulation of visceral fat. Selective celiac angiography showed a pancreatic tumor shadow. Under a diagnosis of insulinoma, the pancreatic body and tail were removed. At 3 months after the operation, the visceral fat area had decreased from 132.6 to 64.2 cm(2). The fasting serum total cholesterol and triglyceride were also reduced. In addition, high-density lipoprotein cholesterol and preheparin serum lipoprotein lipase mass had increased. The midband on the polyacrylamide gel disc electrophoresis of lipoproteins, which appeared before operation, had disappeared completely. An OGTT showed a non-diabetic pattern after the operation. These results suggest that hyperinsulinemia might be one of the factors that enhance visceral adiposity and insulin resistance. PMID- 15356381 TI - Usefulness of serum total cholesterol/triglyceride ratio for predicting the presence of small, dense LDL. AB - OBJECTIVE: We examined the usefulness of the serum total cholesterol (TC)/triglyceride (TG) and LDL-cholesterol (LDL-C)/TG ratios for predicting the presence of small, dense LDL, by comparing them with the established indicators of small, dense LDL, such as the LDL-migration index (LDL-MI) and LDL C/Apolipoprotein B (ApoB) ratio. MATERIALS AND METHODS: Fasting serum lipid was analyzed in 99 Japanese hyperlipidemic and normolipidemic subjects (34 males and 65 females, 59.4 +/- 11.9 years old). RESULTS: A good negative correlation was observed between LDL-MI and log (TC/TG) (R(2) = 0.473, p < 0.0001). There was a strong positive correlation between LDL-C/ApoB and log (TC/TG) (R(2) = 0.665, p < 0.0001). Similar results were obtained using LDL-C instead of TC. Using LDL-MI > 0.4 as an indicator of small, dense LDL, the upper limit of TG was estimated to be 140-142 mg/dl. CONCLUSION: TC/TG and LDL-C/TG may offer a convenient and simple clinical tool for predicting the presence of small, dense LDL. Particularly, TC/TG could be an easy-to-use indicator of small, dense LDL for general practitioners. PMID- 15356382 TI - Relationship between the presence of small, dense low-density lipoprotein and plasma lipid phenotypes in Japanese children. AB - To clarify the relationship between the expression of atherogenic small, dense low-density lipoprotein (SDLDL) and underlying lipid metabolic abnormalities, the prevalence of SDLDL in relation to the serum lipid phenotype was analyzed in 229 children. The LDL particle size was measured using gradient gel electrophoresis, and a particle size of less than 25.5 nm was considered to represent SDLDL. The overall prevalence of SDLDL in the sample population was 8.2% (19/229; 11/117 for boys and 8/112 for girls). Hyperlipidemia phenotype IIb (elevated concentrations of both triglyceride [TG] and total cholesterol [TC]) was strongly associated with SDLDL in 83% (5/6) of the subjects. An elevated TG concentration (phenotype IV) was associated with SDLDL in 55% (10/18) of the subjects. The association between hyperlipidemia phenotype IIa (elevated TC but a normal TG concentration) and SDLDL was quite low (2%; 1/56), but SDLDL was detected in 5% (8/155) of the subjects who presented with normolipidemia. Therefore, these findings suggest that the expression of SDLDL is largely related to lipid abnormalities characterized by phenotype IIb or IV, the underlying metabolic abnormality of which is suspected to be insulin resistance; however, an additional mechanism for the formation of SDLDL that functions independently of plasma lipid abnormalities also seems to exist. PMID- 15356383 TI - Bezafibrate-induced changes over time in the expression of uncoupling protein (UCP) mRNA in the tissues: a study in spontaneously type 2 diabetic rats with visceral obesity. AB - The effect of short-term bezafibrate (BF) administration over time on the expression of UCP mRNA in the tissues was examined in Otsuka Long Evans Tokushima Fatty (OLETF) rats. Eight-week-old rats were divided into a high-dose (100 mg/kg) BF group (n = 15), a low-dose (10 mg/kg) BF group (n = 15) and a control group (n = 15), and followed for 14 days. Feed intake by the high-dose BF group increased significantly between days 10 and 14 of administration. Triglyceride, free fatty acid, and T(4) levels decreased significantly in a dose-dependent manner in the high-dose BF group. Leptin and insulin levels significantly decreased on days 3 and 7. Throughout the study period, liver UCP2 mRNA increased in the high-dose BF group. On day 3 of BF administration, the levels of UCP2 mRNA expression in the skeletal muscles as well as UCP3 mRNA expression in the WAT were significantly increased in the high-dose BF group. PPAR-alpha mRNA significantly increased in the liver on day 3 of BF administration. We thus conclude that the PPAR-alpha mediated effects of BF on the expression of liver UCP2 may be one of the factors that helped to decrease insulin levels. PMID- 15356384 TI - The promoters of two isoflavone synthase genes respond differentially to nodulation and defense signals in transgenic soybean roots. AB - Isoflavonoids are a group of secondary metabolites common to leguminous plants that play roles in nodulation and defense responses. Isoflavone synthase (IFS) catalyzes the key entry point step of isoflavone biosynthesis from the general phenylpropanoid pathway. We have cloned the 5' upstream regions of the genes encoding the two isoflavone synthase isoforms from soybean. We characterized the tissue-specific expression patterns of IFS1 and IFS2 genes of soybean by quantitative RT-PCR. Isoflavone synthase transcripts were detected primarily in the roots and seeds. In transgenic soybean plants in which the IFS1 promoter drove expression of the bacterial uidA (GUS) gene, the expression was localized to the root epidermis and root hairs. We detected differential tissue-specific expression of IFS1:GUS in response to nodulation and defense signals. Bradyrhizobium japonicum, a diazotropic symbiont of soybean, induced the expression of IFS1:GUS in root hairs and xylem poles in the young differentiation zone of the root. This observation suggests novel roles for isoflavonoids in nodulation. The defense related hormone salicylic acid induced the expression of IFS1:GUS in all cell types of the root. We also noticed corresponding alterations in the transcript levels of IFS1 and IFS2 in response to B. japonicum or salicylic acid as identified by quantitative RT-PCR. Using hairy root transformed soybean composite plants, we have identified regions of the IFS1 and IFS2 promoters mediating response to B. japonicum. We have also shown conserved root specific expression of IFS1 in rice and Arabidopsis. The expression pattern of IFS1 in soybean is consistent with the physiological roles of isoflavonoids as defense compounds against pathogens and signal molecules to symbiotic bacteria in soybean. PMID- 15356385 TI - Lhca5--an LHC-type protein associated with photosystem I. AB - The light-harvesting antenna of higher plant photosystem (PS) I is known to be composed of four different types of light-harvesting complex (LHC) proteins (Lhca1-4). However, the genomic sequence of Arabidopsis thaliana contains open reading frames coding for two additional LHC type proteins (Lhca5-6) that are presumably associated with PSI. While Lhca6 might not be expressed at all, ESTs have been detected for the Lhca5 gene in Arabidopsis and a number of other plant species. Here we demonstrate the presence of the Lhca5 gene product in the thylakoid membrane of Arabidopsis as an additional type of Lhca-protein associated with PSI. Lhca5 seems to be regulated differently from the other LHC proteins since Lhca5 mRNA levels increase under high light conditions. Analyses reported here of Lhca5 in plants lacking individual Lhca1-4 proteins show that it is more abundant in plants lacking Lhca1/4, and suggest that it interacts in a direct physical fashion with Lhca2 or Lhca3. We propose that Lhca5 binds chlorophylls in a similar fashion to the other Lhca proteins and is associated with PSI only in sub-stoichiometric amounts. PMID- 15356386 TI - Diversification of genes encoding mei2 -like RNA binding proteins in plants. AB - A predominantly plant-based family of genes encoding RNA binding proteins is defined by the presence of a highly conserved RNA binding motif first described in the mei2 gene of the fission yeast Schizosaccharomyces pombe. In silico analyses reveal nine mei2 -like genes in Arabidopsis thaliana and six in Oryza sativa. These predicted genes group into four distinct clades, based on overall sequence similarity and subfamily-specific sequence elements. In situ analysis show that Arabidopsis genes from one of these clades, TEL1 and TEL2, are specifically expressed in central zone of the shoot apical meristem and the quiescent center of the root apical meristem, suggesting that they may somehow function to maintain indeterminacy in these tissues. By contrast, members of two sister clades, AML1 through AML5, are expressed more broadly, a trend that was confirmed by Q-PCR analysis. mei2 -like transcripts with similar sequences showed similar expression patterns, suggesting functional redundancy within the four clades. Phenotypic analyses of lines that contain T-DNA insertions to individual mei2 -like genes reveal no obvious phenotypes, further suggesting redundant activities for these gene products. PMID- 15356387 TI - Characterization of the Arabidopsis TU8 glucosinolate mutation, an allele of TERMINAL FLOWER2. AB - Glucosinolates are a group of defense-related secondary metabolites found in Arabidopsis and other cruciferous plants. Levels of leaf glucosinolates are regulated during plant development and increase in response to mechanical damage or insect feeding. The Arabidopsis TU8 mutant has a developmentally altered leaf glucosinolate profile: aliphatic glucosinolate levels drop off more rapidly, consistent with the early senescence of the mutant, and the levels of two indole glucosinolates are uniformly low. In TU8 seeds, four long-chain aliphatic glucosinolates have significantly increased levels, whereas the indolyl-3-methyl glucosinolate level is significantly reduced relative to wild type. Genetic mapping and DNA sequencing identified the TU8 mutation as tfl2-6, a new allele of TERMINAL FLOWER2 (TFL2), the only Arabidopsis homolog of animal HETEROCHROMATIN PROTEIN1 (HP1). TU8 (tfl2-6) has other previously identified tfl2 phenotypes, including an early transition to flowering, altered meristem structure, and stunted leaves. Analysis of two additional alleles, tfl2-1 and tfl2-2, showed glucosinolate profiles similar to those of line TU8 (tfl2-6). PMID- 15356388 TI - An arabidopsis promoter microarray and its initial usage in the identification of HY5 binding targets in vitro. AB - To analyze transcription factor-promoter interactions in Arabidopsis, a general strategy for generating a promoter microarray has been established. This includes an integrated platform for promoter sequence extraction and the design of primers for the PCR amplification of the promoter regions of annotated genes in the Arabidopsis genome. A web-interfaced primer-retrieval program was used to obtain up to 10 primer pairs with a suitability ranking given to each gene. We selected primer pairs for the promoters of about 3800 genes, and greater than 95% of the promoter fragments from the total genomic DNA were successfully amplified by PCR. These PCR products were purified and used to print an Arabidopsis promoter microarray. This initial promoter microarray was used to study the in vitro binding of the transcription factor HY5 to its promoter targets. A set of promoter fragments exhibited consistent and strong interaction with the HY5 protein in vitro, and computational analysis revealed that they were enriched with the HY5 consensus binding G-box motif. Thus, a promoter microarray can be a useful tool for identifying transcription factor binding sites at the genomic scale in higher plants. PMID- 15356389 TI - Virus induced gene silencing of a DEFICIENS ortholog in Nicotiana benthamiana. AB - Traditionally, developmental studies in plant biology have suffered from the lack of a convenient means to study gene function in non-model plant species. Here we show that virus-induced gene silencing (VIGS) is an effective new tool to study the function of orthologs of floral homeotic genes such as DEFICIENS (DEF) in non model systems. We used a tobacco rattle virus (TRV)-based VIGS approach to study the function of the Nicotiana benthamiana DEF ortholog (NbDEF). Silencing of NbDEF in N. benthamiana using TRV-VIGS was similar to that of Antirrhinum def and Arabidopsis ap3 mutants and caused transformation of petals into sepals and stamens into carpels. Molecular analysis of the NbDEF -silenced plants revealed a dramatic reduction of the levels of NbDEF mRNA and protein in flowers. NbDEF silencing was specific and has no effect on the mRNA levels of NbTM6, the closest paralog of NbDEF. A dramatic reduction of the levels of N. benthamiana GLOBOSA (NbGLO) mRNA and protein was also observed in flowers of NbDEF-silenced plants, suggesting that cross-regulation of this GLO-like gene by NbDEF. Taken together, our results suggest that NbDEF is a functional homolog of Antirrhinum DEF. Our results are significant in that they show that TRV efficiently induces gene silencing in young and differentiating flowers and that VIGS is a promising new tool for analyses of developmental gene function in non-model organisms. PMID- 15356390 TI - An expression analysis of a gene family encoding plasma membrane aquaporins in response to abiotic stresses in Arabidopsis thaliana. AB - Aquaporin belongs to a highly conserved group of membrane proteins called major intrinsic proteins that facilitate water transport across biological membranes. The genome of Arabidopsis encodes 35 aquaporin genes with 13 homologs in the plasma membrane intrinsic protein (PIP) subgroup. However, the function of each individual aquaporin isoform and the integrated function of plant aquaporins under various physiological conditions remain unclear. As a step toward understanding the aquaporin function in plants under various environmental stimuli, the expressions of a gene family encoding 13 PIPs in Arabidopsis thaliana under various abiotic stress conditions including drought, cold, and high salinity, or abscisic acid (ABA) treatment were investigated by a quantitative real-time reverse transcription-PCR analysis. Several PIP genes were predominantly expressed either in the roots or in the flowers. The expressions of both the highly expressed aquaporins including PIP1;1, PIP1;2, and PIP2;7 and the weakly expressed aquaporins such as PIP1;4, PIP2;1, PIP2;4, and PIP2;5 were modulated by external stimuli. The analyses of our data revealed that only the PIP2;5 was up-regulated by cold treatment, and most of the PIP genes were down regulated by cold stress. Marked up- or down-regulation in PIP expression was observed by drought stress, whereas PIP genes were less-severely modulated by high salinity. The responsiveness of each aquaporin to ABA were different, implying that the regulation of aquaporin expression involves both ABA-dependent and ABA-independent signaling pathways. Together, our comprehensive expression profile of the 13 members of the PIP gene family provides novel basis to allocate the stress-related biological function to each PIP gene. PMID- 15356391 TI - Chromosome walking in the Petunia inflata self-incompatibility (S-) locus and gene identification in an 881-kb contig containing S2-RNase. AB - Self-incompatibility (SI) in the Solanaceae, Rosaceae and Scrophulariaceae is controlled by the polymorphic S locus, which contains two separate genes encoding pollen and pistil determinants in SI interactions. The S-RNase gene encodes the pistil determinant, whereas the pollen determinant gene, named the pollen S gene, has not yet been identified. Here, we set out to construct an integrated genetic and physical map of the S locus of Petunia inflata and identify any additional genes located at this locus. We first conducted chromosome walking at the S2 locus using BAC clones that contained either S2-RNase or one of the nine markers tightly linked to the S locus. Ten separate contigs were constructed, which collectively spanned 4.4 Mb. To identify additional genes located at the S2 locus, a 328-kb region (part of an 881-kb BAC contig) containing S2-RNase was completely sequenced. Approximately 76% of the region contained repetitive sequences, including transposon-like sequences. Other than S2-RNase, an F-box gene, named PiSLF2 (S2-allele of P. inflata S-locus F-box gene), was the only predicted gene whose deduced amino acid sequence was similar to the sequences of known proteins in the database. Two different cDNA selection methods were used to identify additional genes in the 881-kb contig; 11 groups of cDNA clones were identified in addition to those for S2-RNase and PiSLF2. RT-PCR analysis of expression profiles and PCR analysis of BAC clones and genomic DNA confirmed that seven of these 11 newly identified genes were located in the 881-kb contig. PMID- 15356392 TI - Overexpression of multiple dehydrin genes enhances tolerance to freezing stress in Arabidopsis. AB - To elucidate the contribution of dehydrins (DHNs) to freezing stress tolerance in Arabidopsis, transgenic plants overexpressing multiple DHN genes were generated. Chimeric double constructs for expression of RAB18 and COR47 (pTP9) or LTI29 and LTI30 (pTP10) were made by fusing the coding sequences of the respective DHN genes to the cauliflower mosaic virus 35S promoter. Overexpression of the chimeric genes in Arabidopsis resulted in accumulation of the corresponding dehydrins to levels similar or higher than in cold-acclimated wild-type plants. Transgenic plants exhibited lower LT50 values and improved survival when exposed to freezing stress compared to the control plants. Post-embedding immuno electron microscopy of high-pressure frozen, freeze-substituted samples revealed partial intracellular translocation from cytosol to the vicinity of the membranes of the acidic dehydrin LTI29 during cold acclimation in transgenic plants. This study provides evidence that dehydrins contribute to freezing stress tolerance in plants and suggests that this could be partly due to their protective effect on membranes. PMID- 15356393 TI - Isolation and characterization of a rice cysteine protease gene, OsCP1, using T DNA gene-trap system. AB - The T-DNA gene-trap system has been efficiently used to elucidate gene functions in plants. We report here a functional analysis of a cysteine protease gene, OsCP1, isolated from a pool of T-DNA insertional rice. GUS assay with the T-DNA tagged line indicated that the OsCP1 promoter was highly active in the rice anther. Sequence analysis revealed that the deduced amino acid sequence of OsCP1 was homologous to those of papain family cysteine proteases containing the highly conserved interspersed amino acid motif, ERFNIN. This result suggested that the gene encodes a cysteine protease in rice. We also identified a suppressed mutant from T2 progeny of the T-DNA tagged line. The mutant showed a significant defect in pollen development. Taken together, the results demonstrated that OsCP1 is a cysteine protease gene that might play an important role in pollen development. PMID- 15356395 TI - The wired pediatric intensive care unit today: what is your patient's URL? PMID- 15356394 TI - Arabidopsis transcriptional activators CBF1, CBF2, and CBF3 have matching functional activities. AB - When Arabidopsis is exposed to low temperature a small gene family encoding transcription factors known as CBF1, CBF2, and CBF3 (also referred to as DREB1b, DREB1c, and DREB1a, respectively) is rapidly induced followed by expression of CBF-targeted genes, the CBF regulon, which act to bring about an increase in freezing tolerance. The CBF1, 2 and 3 proteins, though highly similar in amino acid sequence, are not identical, raising the question of whether the proteins have the same functions. Here we explored this issue by comparing the effects that overexpression of each CBF gene had on Arabidopsis growth and development, proline and sugar composition, freezing tolerance and gene expression. Taken together, the results support the conclusion that the CBF1, 2 and 3 transcriptional activators have redundant functional activities. PMID- 15356396 TI - Extracorporeal cardiopulmonary resuscitation: ready for prime time? PMID- 15356397 TI - Do we really need more confirmation on the usefulness of inhaled nitric oxide in children's acute respiratory distress syndrome? PMID- 15356398 TI - Bronchiolitis: what's next? PMID- 15356400 TI - Osteoarthritis in women after menopause. PMID- 15356401 TI - Association of breast cancer and its therapy with menopause-related symptoms. PMID- 15356402 TI - It might be wise to consider adding androgen to the estrogen or estrogen progestin regimens in the appropriate patients. PMID- 15356403 TI - Suppression of elevated cartilage turnover in postmenopausal women and in ovariectomized rats by estrogen and a selective estrogen-receptor modulator (SERM). AB - OBJECTIVE: Several observational studies indicate that estrogen deficiency increases the incidence of osteoarthritis in postmenopausal women. To validate this observation, we investigated the effects of ovariectomy (OVX) on cartilage erosion in rats using histology and an established bio-assay of cartilage specific collagen type II degradation products (CTX-II). Furthermore, we investigated whether estrogen and levormeloxifene, a selective estrogen-receptor modulator (SERM), can prevent the OVX-induced changes in cartilage degradation. The clinical relevance was assessed in postmenopausal women by measuring the changes in CTX-II during 12-month treatment with levormeloxifene versus placebo. DESIGN: Sixty 6-month-old rats were divided in five groups. One group was subjected to sham and the others to OVX, followed by treatment with vehicle alone, estradiol or 0.2 mg/kg/day or 5 mg/kg/day of levormeloxifene. The rats were treated for 9 weeks with biweekly blood and urine sampling for measurement of bone resorption and cartilage turnover. After study termination, hind knees were removed for histological analysis of erosions. The effect of levormeloxifene in post-menopausal women was assessed by measuring CTX-II in samples from 301 women who were participating in a phase II study of this SERM. RESULTS: OVX rats showed significant increases in the urinary excretion of CTX-II. After 9 weeks this was manifested as increased surface erosion of knee articular cartilage compared with sham-operated rats. Treatment with estrogen or levormeloxifene prevented the OVX-induced changes. There was a significant correlation between the 4-week changes in CTX-II and cartilage erosion at week 9 (r = 0.64, P < 0.001). In postmenopausal women treated with levormeloxifene, the urinary excretion of CTX-II was decreased by approximately 50% and restored CTX-II levels to the premenopausal range. CONCLUSIONS: This study is the first to demonstrate that a SERM suppresses cartilage degradation in both rodents and humans, suggesting potential therapeutical benefits in the prevention of destructive joint diseases such as osteoarthritis. PMID- 15356404 TI - Association of breast cancer and its therapy with menopause-related symptoms. AB - OBJECTIVE: To study the relationship between current menopause status, occurrence of menopause transition during cancer treatment, and prevalence and severity of possible menopause-related symptoms. DESIGN: Data from the Cancer and Menopause Study (CAMS), a tumor-registry-based cohort of breast cancer survivors (BCS) diagnosed before age 50, were used. Using a standardized symptom checklist, women reported whether they were not at all, slightly, moderately, quite, or extremely bothered in the past 4 weeks by hot flashes, night sweats, vaginal dryness, pain with intercourse, breast sensitivity, joint pains, frequent mood changes, restless sleep, weight gain, forgetfulness, and difficulty concentrating. Current menopause status (by standard categories based on menstruation) and whether a persistent menopause transition occurred during cancer treatment were the main exposures. Linear (symptom severity as continuous outcome) and logistic (symptom present vs absent) regression models were adjusted for age, ethnicity, current smoking, alcohol use, chemotherapy, tamoxifen, body mass index, and depression scores. RESULTS: Mean age of the participants was 50 years. The prevalence of symptoms was high. Hot flashes occurred in 17%, 51%, and 71% of pre-, peri-, and postmenopausal BCS, respectively. Hot flashes, vaginal dryness, and pain with intercourse were more severe in postmenopausal compared with perimenopausal BCS. Having had a transition during breast cancer treatment was associated with worse hot flash severity, independent of current menopause status. CONCLUSIONS: Menopause-related symptoms are common in BCS. Effective treatment options are needed. Having a treatment-related transition confers a persistent effect on hot flash severity. Clinicians should include this information when counseling women on potential outcomes of their cancer therapy. PMID- 15356405 TI - Breast cancer incidence in postmenopausal women using testosterone in addition to usual hormone therapy. AB - OBJECTIVE: There is now convincing evidence that usual hormone therapy for ovarian failure increases the risk for breast cancer. We have previously shown that ovarian androgens normally protect mammary epithelial cells from excessive estrogenic stimulation, and therefore we hypothesized that the addition of testosterone to usual hormone therapy might protect women from breast cancer. DESIGN: This was a retrospective, observational study that followed 508 postmenopausal women receiving testosterone in addition to usual hormone therapy in South Australia. Breast cancer status was ascertained by mammography at the initiation of testosterone treatment and biannually thereafter. The average age at the start of follow-up was 56.4 years, and the mean duration of follow-up was 5.8 years. Breast cancer incidence in this group was compared with that of untreated women and women using usual hormone therapy reported in the medical literature and to age-specific local population rates. RESULTS: There were seven cases of invasive breast cancer in this population of testosterone users, for an incidence of 238 per 100,000 woman-years. The rate for estrogen/progestin and testosterone users was 293 per 100,000 woman-years--substantially less than women receiving estrogen/pro-gestin in the Women's Health Initiative study (380 per 100,000 woman-years) or in the "Million Women" Study (521 per 100,000 woman years). The breast cancer rate in our testosterone users was closest to that reported for hormone therapy never-users in the latter study (283 per 100,000 woman-years), and their age-standardized rate was the same as for the general population in South Australia. CONCLUSIONS: These observations suggest that the addition of testosterone to conventional hormone therapy for postmenopausal women does not increase and may indeed reduce the hormone therapy-associated breast cancer risk-thereby returning the incidence to the normal rates observed in the general, untreated population. PMID- 15356406 TI - Health behaviors and other characteristics of women on hormone therapy: results from the Third National Health and Nutrition Examination Survey, 1988-1994. AB - OBJECTIVE: To examine the prevalence of hormone therapy (HT) use and compare demographic characteristics, health behaviors, and health indicators between current HT users and never-users in a nationally representative sample of postmenopausal women. DESIGN: The Third National Health and Nutrition Examination Survey was a cross-sectional survey conducted between 1988 and 1994, including 3,673 postmenopausal women aged 40 years and older. RESULTS: Overall, 419 (11.4%) of the women reported current HT use, 857 (23.3%) reported past use, and 2,397 (65.3%) were never-users. Non-Hispanic black women and women aged 70 years or older were less likely to be current users. Higher socioeconomic status (education and income) and surgical menopause were associated with increased odds of current hormone use. After adjusting for the above variables, women who reported being inactive during leisure time and obese women (body mass index >or= 30) were less likely to be current users. Women who had 5 to 29 alcoholic drinks per month, perceived their health status as "good," took a multiple vitamin, were aware of having high blood cholesterol, and had a clinic for regular medical care were more likely to be current users. Smoking habits were not significantly different between the groups. CONCLUSIONS: Current HT users have different demographic profiles and may lead healthier lives than never-users. This is important to take into account when studying the effects of HT, and it may partly explain differences in findings regarding the health effects of HT use in observational studies compared with randomized clinical trials. PMID- 15356407 TI - Predicting participation in prospective studies of ovarian aging. AB - OBJECTIVE: Identifying clinical markers and characteristics of the transition to menopause is an important woman's health issue, and recent long-term, prospective, cohort studies are just beginning to offer insight into methods of predicting the transition to menopause. One of the major challenges of conducting prospective cohort studies is the problem of attrition-both the retention of study participants and the influence of dropouts on the final study results. We conducted this systematic analysis to: 1) identify baseline predictors of subsequent long-term participation, and 2) determine the demographic, psychosocial, and hormonal differences between participants and dropouts among a group of premenopausal women enrolled in a longitudinal study of ovarian aging. DESIGN: Using data from the Penn Ovarian Aging study, premenopausal women aged 35 to 47 enrolled in the study were classified as either Active (full participants), Skipped, or Dropped participants based on their visit pattern during a 4-year study interval. RESULTS: Most of the demographic and psychosocial variables did not significantly differ between the Active, Skipped, or Dropped groups. There was no racial difference in study participation. The Dropouts were more likely to have a high school education and were less likely to report menopause symptoms compared with the Actives (P < 0.01). The Skipped group reported more anxiety (P < 0.05), and members were more likely to have less than a high school education compared with the Actives (P < 0.03). Hormone levels (estradiol, follicle stimulating hormone, luteinizing hormone, dehydroepiandrosterone, testosterone) at enrollment were within the premenopausal range and did not significantly differ among the three study groups. These findings remained after adjustment for covariates and hormone levels in multivariate analyses. CONCLUSIONS: Education, anxiety levels and menopause symptoms at baseline differed marginally between the women participating fully and those who dropped out or skipped multiple assessments. These findings are important and indicate that long-term study participation rates do not differ substantially by racial group or any of the other demographic or hormonal characteristics examined. PMID- 15356408 TI - Combined hormone therapy in postmenopausal women with features of metabolic syndrome. Results from a population-based study of Swedish women: Women's Health in the Lund Area study. AB - OBJECTIVE: To delineate the influence of hormone therapy (HT) on features of metabolic syndrome with special reference to the composition and mode of administration of three specific HT regimens, all containing estradiol (E2) + norethisterone. DESIGN: The Women's Health in the Lund Area project screened all women (n = 10,766), born between 1935 and 1945. Complete data were obtained from 6,917 women. Those at or above defined cutoff limits were considered positively screened (n = 3,593) for metabolic syndrome. All of them were invited to undergo an oral glucose tolerance test; 2,923 women accepted. After excluding 200 women with impaired fasting glucose, 2,723 women were included in the present analysis. Serum lipids were determined by conventional standard methods at the department of clinical chemistry of Lund University Hospital. RESULTS: According to World Health Organization criteria, 2,123 women had normal glucose tolerance and 600 women had impaired glucose tolerance (IGT). IGT was less common (P = 0.001) among users of a transdermal patch [CYC-TRANS; E2 50 microg + norethisterone acetate (NETA) 250microg] compared with the two-combined oral regimen [CON-O (continuous oral E2 2 mg + NETA 1 mg) + CYC-O (sequential oral E2 2 mg + NETA 1 mg)]. Furthermore, IGT was more common among CON-O users when compared with either the CYC-O + CYC-TRANS group (P = 0.002) or the CYC-TRANS only group (P = 0.001). There were no significant differences between CYC-O versus CYC-TRANS or CON-O. Serum levels of total cholesterol were higher in the CYC-TRANS group than in the combined CON-O + CYC-O group (P < 0.05); they also were higher (P = 0.05) when comparing the CYC-O + CYC-TRANS versus CON-O as well as higher in CYC-TRANS versus CON-O (P < 0.05). Serum high-density lipoprotein cholesterol levels were higher in the CYC-O (P = 0.001), CYC-TRANS (P < 0.05), and the CYC-O + CYC-TRANS (P = 0.001) groups when compared with the CON-O users. There were no differences in the mean age, blood pressure (systolic and diastolic), body mass index, waste hip ratio, or the rate of cigarette and alcohol consumption between the different hormone regimens. CONCLUSION: The risk of having a pathological glucose load was lower in transdermal versus oral users of HT. Transdermal HT could be regarded as first-line treatment in women at risk of developing diabetes. PMID- 15356409 TI - Effect of estrogen use on tooth retention, oral bone height, and oral bone porosity in Japanese postmenopausal women. AB - OBJECTIVE: Recent studies in the United States support the protective effect of estrogen use on tooth retention; however, little is known as to how estrogen promotes tooth retention. The aims of this study were to investigate the effects of estrogen use on tooth retention, oral bone height, and oral bone porosity in Japanese postmenopausal women and to clarify how estrogen promotes tooth retention. DESIGN: Relationships among the number of teeth remaining (total, anterior, and posterior teeth), oral bone height, oral bone porosity, bone mineral density of the lumbar spine and the femoral neck, estrogen use status, and the duration of estrogen use were evaluated in 330 Japanese postmenopausal women (mean age +/- SD, 56.8 +/- 7.6 y). RESULTS: Analysis of covariance adjusted for confounding variables revealed that estrogen users (66 women) tended to have more posterior teeth than did nonusers (264 women) (P = 0.065), although there were no significant differences in number of total (P = 0.196) and anterior (P = 0.751) teeth remaining, oral bone height (P = 0.970), oral bone porosity (P = 0.745), and bone mineral density of the lumbar spine (P = 0.459) and the femoral neck (P = 0.749) between estrogen users and nonusers. Multiple regression analysis showed that the duration of estrogen use was significantly associated with number of total (P = 0.019) and posterior (P = 0.007) teeth remaining, independent of age and oral bone height. CONCLUSION: Our results suggest that estrogen may promote tooth retention by strengthening the periodontal attachment surrounding the teeth, but not increasing oral bone height and not decreasing oral bone porosity. PMID- 15356410 TI - A population-based study of depressed mood in middle-aged, Australian-born women. AB - OBJECTIVE: There has been controversy about the relationship between menopause and depression. This study utilizes a unique prospective population-based data set of middle-aged, Australian-born women to identify determinants of depressed mood. DESIGN: The Melbourne Women's Midlife Health Project sample consisted of 438 women aged 45 to 55 at baseline; they were followed annually for 11 years. Of this group, 314 (72%) completed the Center for Epidemiologic Studies Depression Scale (CES-D) scale in year 11 to measure depressed mood. Variables measured at baseline and annually included negative mood (measured with Affectometer) and psychosocial, hormonal, health, and lifestyle factors. RESULTS: Women who had the highest CES-D scores were those who by year 11 were still in the menopause transition stage (had not reached final menstrual period) or had experienced surgical menopause. CES-D correlated with negative mood measured concurrently (r = 0.63) and baseline negative mood (r = 0.37). There was a significant reduction in negative mood for all menopause status groups, but those who experienced surgical menopause showed less reduction than other women. Ever-use or number of years of use of hormone therapy made no difference to CES-D outcome. CES-D was associated with baseline negative attitudes toward aging, mood, and premenstrual complaint experience and annual mood, poor self-rated health, number of bothersome symptoms, and daily hassles. CONCLUSIONS: Women most likely to have higher depressed mood in the age group 57 to 67 are those who have undergone surgical menopause or have menstruated within the last 12 months. Prior negative mood, history of premenstrual complaints, negative attitudes toward aging or menopause, poor health, and daily hassles predict depressed mood. PMID- 15356411 TI - Regional differences in age at menopause between Korean-Korean and Korean Chinese. AB - OBJECTIVE: To evaluate the difference in age at menopause between Korean-Korean (KK) women and Korean-Chinese (KC) women. DESIGN: A total of 1,972 participants were recruited: 961 postmenopausal women living in Korea, and 1,011 second- or third-generation emigrants to China. A structured questionnaire was used that included current age (at interview), age at menopause, weight, height, duration of education, socioeconomic status (SES), smoking, alcohol intake, number of children and abortions, etc. For every independent variable, the effect of regional difference (ie, living in Korea or China) on age at menopause was analyzed. Student's t test and chi2 test were applied as appropriate, and multiple regression analyses were performed for significant variables. RESULTS: Age at menopause was significantly higher in KK than KC (49.3 +/- 3.5 y vs 48.9 +/- 3.1 y, P = 0.011). Body mass index (BMI) and number of abortions were also higher in KK (23.3 +/- 2.7 vs 23.1 +/- 2.6, P = 0.021; 1.3 +/- 1.7 vs 0.2 +/- 0.7, P = 0.0001). Duration of education was significantly longer in KK. Smokers showed a significantly earlier menopause onset. BMI was significantly and positively correlated with age at menopause, whereas current SES was negatively correlated. When regional differences were assessed in each categorized subgroup, age at menopause showed a significant difference, too. After adjusting for age, SES, BMI, smoking, and number of children, onset of menopause was still earlier in KC (P = 0.0001, R = 0.037). CONCLUSION: KK women were found to have a later onset of menopause than KC women, and our results suggest that, apart from the well-known genetic influence, environmental effects play a significant role in determining age at menopause. PMID- 15356412 TI - Autoimmune hepatitis associated with the use of black cohosh: a case study. AB - Herbal remedies generate more than 1.8 billion dollars in annual sales in the United States. Herbal products have been associated with a wide spectrum of hepatic toxicities. With the recent Women's Health Initiative Study demonstrating increased risk of breast cancer and cardiovascular events associated with hormone therapy, many women may resort to herbal remedies for persistent menopause symptoms. We report a case of autoimmune hepatitis likely triggered by the use of black cohosh (Actaea racemosa), an agent marketed to treat menopause symptoms. Given this case report, we recommend close monitoring of women using this herbal preparation. PMID- 15356413 TI - A perspective upon the menopausal transition impossible a decade ago currently appears to be in process of formation. PMID- 15356414 TI - The Atypical Antipsychotic Therapy and Metabolic Issues National Survey: practice patterns and knowledge of psychiatrists. AB - A nationwide survey in 2003 of 300 randomly selected psychiatrists who routinely treat schizophrenia with atypical antipsychotic therapy was conducted to ascertain practice patterns and attitudes regarding metabolic disturbances during atypical antipsychotic therapy with an emphasis on how these perceptions impact therapeutic decision making. Psychiatrists generally believe that some atypical antipsychotic drugs are associated with metabolic disturbances and that atypical antipsychotics differ in their risk for metabolic disturbances. A majority of respondents (82%) believed that patients with schizophrenia-even those not receiving atypical antipsychotic therapy-are at greater risk for metabolic abnormalities than the general population. A majority of respondents recognized weight gain and diabetes mellitus (59% and 51%, respectively) as potential metabolic complications of atypical antipsychotic therapy, while only some recognized dyslipidemia and certain acute metabolic decompensations like diabetic ketoacidosis (22% and 2%, respectively). Large minorities of respondents (48% and 43%) indicated a willingness to risk weight gain and/or diabetes for the benefits of atypical antipsychotics, possibly because metabolic issues were regarded as long-term issues. However, large majorities also stated that they considered metabolic issues when selecting atypical antipsychotic therapy for some or all of their patients (90%), and that emergence of metabolic dysfunction prompted them to change atypical antipsychotic treatment regimens (85%). Additional efforts at continuing education and communication regarding metabolic outcomes associated with atypical antipsychotic therapy, as well as critical reviews in this area, may help clarify atypical antipsychotic treatment risks and benefits. The results from the survey indicate that psychiatrists are aware of and concerned about metabolic risks and how they differ across the atypical antipsychotic class. The impact of additional data and educational efforts in this area, such as a recently published consensus statement from the American Diabetes Association and other organizations, remains to be assessed. PMID- 15356415 TI - Atypical antipsychotics and metabolic dysregulation: evaluating the risk/benefit equation and improving the standard of care. AB - Atypical antipsychotics are a major advance in the management of schizophrenia. The reevaluation of widely held risk/benefit assessments of the various atypical antipsychotics provides an opportunity to improve treatment patterns. The best available clinical trial evidence indicates that efficacy among the atypical antipsychotics (at equivalent doses) is very similar, but safety and tolerability profiles differ significantly. Atypical antipsychotics differ markedly in their potential to cause metabolic disturbances, including obesity, diabetes, dyslipidemia, and the metabolic syndrome; clozapine and olanzapine carry the greatest risks, atypical antipsychotics like risperidone and quetiapine have lower risks, and newer agents like ziprasidone and aripiprazole are associated with minimal metabolic risks. Results from the Atypical Antipsychotic Therapy and Metabolic Issues (AtAMI) survey define important opportunities for improving medical and psychiatric outcomes during atypical antipsychotic therapy. (See accompanying article by Newcomer et al) Additional educational and research efforts are required to increase understanding of common conditions such as the metabolic syndrome, increase awareness of uncommon but serious events like diabetic ketoacidosis, and pancreatitis, and identify appropriate strategies for monitoring the risks/benefits of atypical antipsychotic therapy. As clinicians refine practice patterns regarding the atypical antipsychotics, they may require additional knowledge and resources to fully incorporate risk/benefit considerations and optimize long-term psychiatric and medical outcomes. PMID- 15356416 TI - The neurotoxicity of tissue plasminogen activator? AB - Tissue plasminogen activator (tPA), a fibrin specific activator for the conversion of plasminogen to plasmin, stimulates thrombolysis and rescues ischemic brain by restoring blood flow. However, emerging data suggests that under some conditions, both tPA and plasmin, which are broad spectrum protease enzymes, are potentially neurotoxic if they reach the extracellular space. Animal models suggest that in severe ischemia with injury to the blood brain barrier (BBB) there is injury attributed to the protease effects of this exogenous tPA. Besides clot lysis per se, tPA may have pleiotropic actions in the brain, including direct vasoactivity, cleaveage of the N-methyl-D-aspartate (NMDA) NR1 subunit, amplification of intracellular Ca++ conductance, and activation of other extracellular proteases from the matrix metalloproteinase (MMP) family, e.g. MMP 9. These effects may increase excitotoxicity, further damage the BBB, and worsen edema and cerebral hemorrhage. If tPA is effective and reverses ischemia promptly, the BBB remains intact and exogenous tPA remains within the vascular space. If tPA is ineffective and ischemia is prolonged, there is the risk that exogenous tPA will injure both the neurovascular unit and the brain. Methods of neuroprotection, which prevent tPA toxicity or additional mechanical means to open cerebral vessels, are now needed. PMID- 15356417 TI - PAR-1 deficiency protects against neuronal damage and neurologic deficits after unilateral cerebral hypoxia/ischemia. AB - Cardiovascular and neurologic surgeries often involve a temporary reduction in cerebral blood flow. In these conditions, as well as during cerebral ischemia and traumatic brain injury, the temporary loss of oxygen and glucose initiates a cascade of cellular events that culminate in neuronal death and damage. Understanding the mechanisms that contribute to neuronal death after hypoxia/ischemia is critically important for treatment of such brain injury. Here, we use a model of combined cerebral hypoxia/ischemia (H/I) to examine the role of protease-activated receptor-1 (PAR-1) in hypoxic/ischemic neuronal damage. Our data show that PAR-1-deficient mice have smaller lesion volumes than wild-type controls after 45 minutes of H/I. The results of the genetic block of PAR-1 were corroborated using a PAR-1 antagonist, which decreased infarct volume in wild-type C57Bl6 mice. Examination of cellular responses to H/I reveals that PAR-1 -/- animals have less cellular death and diminished glial fibrillary acidic protein expression. Additionally, PAR-1 -/- mice exhibit less motor behavior impairment in rotorod and inverted wire-hang tests. These data suggest that PAR-1 contributes to hypoxic/ischemic brain injury and are consistent with other studies that implicate serine proteases and their receptors in neuropathology after cerebral insults. PMID- 15356418 TI - Glutamatergic neurotransmission and neuronal glucose oxidation are coupled during intense neuronal activation. AB - 13C nuclear magnetic resonance (NMR) experiments have previously shown that glutamatergic neurotransmitter flux (Vcycle(Glu/Gln)) changes proportionately with neuronal glucose oxidation (CMRglc(ox)N) in the nonactivated cortex of anesthetized rats. Positron Emission Tomography measurements of glucose and oxygen uptake during sensory stimulation had shown that the incremental glucose utilization is greater than oxygen leading to the suggestion that the energy required for stimulated neuronal activity arises from nonoxidative glucose metabolism. In this study, the authors used spatially localized 1H-observed, 13C edited NMR spectroscopy during an infusion of [1,6-13C2]glucose to assess the relationship between changes in Vcycle(Glu/Gln) and glucose utilization (CMRglc(ox)N and CMRglc(nonox)) during the intense cortical activity associated with bicuculline-induced seizures. Metabolic fluxes were determined by model based analysis of the 13C-enrichment time courses of glutamate-C4 and glutamine C4 (CMRglc(ox)N, Vcycle(Glu/Gln)) and lactate-C3 (CMRglc(nonox)). The exchange rate between alpha-ketoglutarate and glutamate was found to be significantly faster than TCA cycle flux both for control (41 micromol.g(-1).min(-1); 95% CI, 5 to 109 micromol.g(-1).min(-1)) and during seizures (21 micromol.g(-1).min(-1); 95% CI, 4.4 to 51.8 micromol.g(-1).min(-1)). During seizures, total glucose utilization (CMRglc(ox+nonox)) increased substantially (466% between 0 and 6 minutes; 277% between 6 and 55 minutes). Glucose oxidation (CMRglc(ox)N) also increased (214%; from 0.26 +/- 0.02 to 0.57 +/- 0.07 micromol.g(-1).min(-1)) but to a lesser degree, resulting in a large increase in cortical lactate concentration. Vcycle(Glu/Gln) increased 233% (from 0.22 +/- 0.04 to 0.52 +/- 0.07 micromol.g(-1).min(-1)), which was similar to the increase in glucose oxidation. The value of Vcycle(Glu/Gln) and CMRglc(ox)N obtained here lie on the line predicted in a previous study. These results indicate that neuronal glucose oxidation and not total glucose utilization is coupled to the glutamate/glutamine cycle during intense cortical activation. PMID- 15356419 TI - Mitochondria consume energy and compromise cellular membrane potential by reversing ATP synthetase activity during focal ischemia in rats. AB - The direction of the chemical reaction of ATP synthetase is reversible. The present study was designed to determine whether mitochondria produce or consume ATP during ischemia. For this purpose, changes in mitochondrial membrane potential were measured in vivo at the site of a direct current (DC) electrode using a potentiometric dye, 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolylcarbocyanine iodide (JC-1), and a rat model of focal ischemia. Two microL of dye (control group) or dye with oligomycin, an ATP synthetase inhibitor (oligomycin group), was injected into the parietotemporal cortex through the DC electrode. With the initiation of ischemia, a decrease in mitochondrial potential was observed within 20 seconds in the oligomycin group (earlier than the onset of DC deflection, P = 0.02). In contrast, in the control group, mitochondrial potential was maintained at 91 +/- 5% of the preischemia level for 118 +/- 38 seconds before showing full depolarization simultaneously with DC deflection. During the period of ischemia, the mitochondrial potential was higher in the control group (66 +/- 9%) than in the oligomycin group (46 +/- 8%, P = 0.0002), whereas DC potential was lower in the control group (-18 +/- 3) than in the oligomycin group (-15 +/- 2 mV, P = 0.04). These observations suggest that mitochondria consume ATP during ischemia by reversing ATP synthetase activity, which compromises cellular membrane potential by consuming ATP. PMID- 15356420 TI - Fluorometric determination of glucose utilization in neurons in vitro and in vivo. AB - Glucose is the major energy source the adult brain utilizes under physiologic conditions. Recent findings, however, have suggested that neurons obtain most of their energy from the oxidation of extracellular lactate derived from astroglial metabolism of glucose transported into the brain from the blood. In the present studies we have used 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D glucose (2-NBDG), a fluorescent analogue of 2-deoxyglucose, which is often used to trace glucose utilization in neural tissues, to examine glucose metabolism in neurons in vitro and in vivo. Cultured neurons and astroglia were incubated with 2-NBDG for up to 15 minutes, and nonmetabolized 2-NBDG was washed out. We found that fluorescence intensity increased linearly with incubation time in both neurons and astroglia, indicating that both types of brain cells could utilize glucose as their energy source in vitro. To determine if the same were true in vivo, Sprague-Dawley rats were injected intravenously with a pulse bolus of 2 NBDG and decapitated 45 minutes later. Examination of brain sections demonstrated that phosphorylated 2-NBDG accumulated in hippocampal neurons and cerebellar Purkinje cells, indicating that neurons can utilize glucose in vivo as energy source. PMID- 15356421 TI - Selective uptake of [14C]2-deoxyglucose by neurons and astrocytes: high resolution microautoradiographic imaging by cellular 14C-trajectography combined with immunohistochemistry. AB - At the moment, there is no direct in vivo evidence of the relative amount of glucose taken up and metabolized by glial cells and neurons, respectively. Therefore, we developed a specific high cellular resolution beta-trajectory approach that allows recording and identification of individual tracks of electrons emitted during disintegrations of 14C. We used [14C]2-deoxyglucose (2DG), which is an analog of glucose and is not metabolized further than the first phosphorylation by hexokinase; this property allows localization of the tracer within the cell type where it is phosphorylated. The present technical approach associated a method of cellular trajectography mainly characterized by the high thickness of the emulsion (15 microm), which permits following of the trajectory of individual electrons. This technique was improved to preserve the in vivo label of diffusible compounds such as 2DG and 2DG-6P and associated with immunohistochemical detection of neurons and astrocytes. beta-Track counting of labeled compounds was performed in 5 microm glial fibrillary acidic protein (GFAP)- and microtubule-associated protein (MAP)2-immunolabeled paraffin adjacent sections. Of 3,075 counted beta-tracks, 53.0% were localized in astrocytes on GFAP-labeled sections and 60.1% in neurons on MAP2-labeled sections. These data represent the first in vivo evidence of the compartmentation of uptake and metabolism of glucose in neurons and astrocytes. PMID- 15356422 TI - In vivo measurement of glucose utilization in rats using a beta-microprobe: direct comparison with autoradiography. AB - A new beta-microprobe (betaP) has been used to locally measure the time concentration curve of a radiolabeled substance. The betaP, analogous to positron emission tomography methodology, is useful for in vivo animal studies because it can acquire time-concentration curves with high temporal and spatial resolution. Using [18F]fluoro-2-deoxy-D-glucose and betaP, we evaluated the reliability of the biologic parameters and we compared this method with the [14C]2-deoxy-D glucose autoradiography. BetaP time-concentration curves in three regions of the brain were obtained from 24 rats. Four kinetic parameters (K1-k4) were estimated from 60-minute experimental periods using a three-compartment model. Best fits were obtained when the vascular fraction (Fv) was estimated simultaneously with the four kinetic parameters (K1-k4). The mean estimated Fv values were about 5.5% for the frontal cortex regions and 8.0% for the cerebellum. Correlation coefficients higher than 0.830 were observed between regional cerebral metabolic rates for glucose (rCMRglc) values obtained by betaP and autoradiography. In addition, the betaP-derived input function was similar to that obtained by manual sampling. Our findings show that reliable rCMRglc values can be obtained using betaP. PMID- 15356423 TI - Relationship between flow-metabolism uncoupling and evolving axonal injury after experimental traumatic brain injury. AB - Blood flow-metabolism uncoupling is a well-documented phenomenon after traumatic brain injury, but little is known about the direct consequences for white matter. The aim of this study was to quantitatively assess the topographic interrelationship between local cerebral blood flow (LCBF) and glucose metabolism (LCMRglc) after controlled cortical impact injury and to determine the degree of correspondence with the evolving axonal injury. LCMRglc and LCBF measurements were obtained at 3 hours in the same rat from 18F-fluorodeoxyglucose and 14C iodoantipyrine coregistered autoradiographic images, and compared to the density of damaged axonal profiles in adjacent sections and in an additional group at 24 hours using beta-amyloid precursor protein (beta-APP) immunohistochemistry. LCBF was significantly reduced over the ipsilateral hemisphere by 48 +/- 15% compared with sham-controls, whereas LCMRglc was unaffected, apart from foci of elevated LCMRglc in the contusion margin. Flow-metabolism was uncoupled, indicated by a significant 2-fold elevation in the LCMRglc/LCBF ratio within most ipsilateral structures. There was a significant increase in beta-APP-stained axons from 3 to 24 hours, which was negatively correlated with LCBF and positively correlated with the LCMRglc/LCBF ratio at 3 hours in the cingulum and corpus callosum. Our study indicates a possible dependence of axonal outcome on flow-metabolism in the acute injury stage. PMID- 15356424 TI - [18F]altanserin binding to human 5HT2A receptors is unaltered after citalopram and pindolol challenge. AB - The aim of the present study was to develop an experimental paradigm for the study of serotonergic neurotransmission in humans using positron emission tomography and the 5-HT2A selective radioligand [18F]altanserin. [18F]altanserin studies were conducted in seven subjects using the bolus/infusion approach designed for attaining steady state in blood and brain 2 hours after the initial [18F]altanserin administration. Three hours after commencement of radiotracer administration, 0.25 mg/kg of the selective serotonin reuptake inhibitor, citalopram (Lundbeck, Valby, Denmark), was administered to all subjects as a constant infusion for 20 minutes. To reduce 5-HT1A-mediated autoinhibition of cortical 5-HT release, four of the seven subjects were pretreated with the partial 5-HT1A agonist pindolol for 3 days at an increasing oral dose (25 mg on the day of scanning). In each subject, the baseline condition (120 to 180 minutes) was compared with the stimulated condition (195 to 300 minutes). Despite a pronounced increase in plasma prolactin and two subjects reporting hot flushes compatible with an 5-HT-induced adverse effect, cortical [18F]altanserin binding was insensitive to the citalopram challenge, even after pindolol pretreatment. The biochemical and cellular events possibly affecting the unsuccessful translation of the citalopram/pindolol challenge into a change in 5-HT2A receptor binding of [18F]altanserin are discussed. PMID- 15356425 TI - Bumetanide inhibition of the blood-brain barrier Na-K-Cl cotransporter reduces edema formation in the rat middle cerebral artery occlusion model of stroke. AB - Increased transport of Na+ across an intact blood-brain barrier (BBB) participates in edema formation during the early hours of cerebral ischemia. In previous studies, the authors showed that the BBB Na-K-Cl cotransporter is stimulated by factors present during ischemia, suggesting that the cotransporter may contribute to the increased brain Na+ uptake in edema. The present study was conducted to determine (1) whether the Na-K-Cl cotransporter is located in the luminal membrane of the BBB, and (2) whether inhibition of the BBB cotransporter reduces brain edema formation. Perfusion-fixed rat brains were examined for cotransporter distribution by immunoelectron microscopy. Cerebral edema was evaluated in rats subjected to permanent middle cerebral artery occlusion (MCAO) by magnetic resonance diffusion-weighted imaging and calculation of apparent diffusion coefficients (ADC). The immunoelectron microscopy studies revealed a predominant (80%) luminal membrane distribution of the cotransporter. Magnetic resonance imaging studies showed ADC ratios (ipsilateral MCAO/contralateral control) ranging from 0.577 to 0.637 in cortex and striatum, indicating substantial edema formation. Intravenous bumetanide (7.6-30.4 mg/kg) given immediately before occlusion attenuated the decrease in ADC ratios for both cortex and striatum (by 40-67%), indicating reduced edema formation. Bumetanide also reduced infarct size, determined by TTC staining. These findings suggest that a luminal BBB Na-K-Cl cotransporter contributes to edema formation during cerebral ischemia. PMID- 15356426 TI - Relative changes in cerebral blood flow and neuronal activity in local microdomains during generalized seizures. AB - There is broad agreement that generalized tonic-clonic seizures (GTCS) and normal somatosensory stimulation are associated with increases in regional CBF. However, the data regarding CBF changes during absence seizures are controversial. Electrophysiologic studies in WAG/Rij rats, an established animal model of absence seizures, have shown spike-wave discharges (SWD) that are largest in the perioral somatosensory cortex while sparing the visual cortex. Recent functional magnetic resonance imaging (fMRI) studies in the same model have also shown localized increases in fMRI signals in the perioral somatosensory cortex during SWD. Because fMRI signals are only indirectly related to neuronal activity, the authors directly measured CBF and neuronal activity from specific microdomains of the WAG/Rij cortex using a specially designed probe combining laser-Doppler flowmetry and extra-cellular microelectrode recordings under fentanyl/haloperidol anesthesia. Using this approach, parallel increases in neuronal activity and CBF were observed during SWD in the whisker somatosensory (barrel) cortex, whereas the visual cortex showed no significant changes. For comparison, these measurements were repeated during somatosensory (whisker) stimulation, and bicuculline-induced GTCS in the same animals. Interestingly, whisker stimulation increased neuronal activity and CBF in the barrel cortex more than during SWD. During GTCS, much larger increases that included both the somatosensory and visual cortex were observed. Thus, SWD in this model produce parallel localized increases in neuronal activity and CBF with similar distribution to somatosensory stimulation, whereas GTCS produce larger and more widespread changes. The normal response to somatosensory stimulation appears to be poised between two abnormal responses produced by two physiologically different types of seizures. PMID- 15356427 TI - Drug trial registries. PMID- 15356428 TI - Vesnarinone-mediated alterations of gene expression in cardiac fibroblasts from aortic regurgitant hearts. AB - Pathologic fibrosis precedes heart failure (CHF) and death in experimental aortic regurgitation (AR). Vesnarinone, a positively inotropic quinolone derivative, suppresses survival of fibroblasts (CF) from hearts with chronic experimental AR. To explore further the potential effects of vesnarinone on cardiac fibrosis in AR, we tested the hypothesis that vesnarinone suppresses gene expression induced by AR in CF. Differentially expressed genes were isolated by suppression subtractive hybridization (SSH) in CF from hearts of 2 New Zealand White rabbits with surgically induced AR compared with 2 normal rabbits. In cultured AR-CF treated with and without vesnarinone (4 doses, including the dose that had caused maximal survival suppression in cultured AR-CF), drug effect was assessed on expression of genes found to be up-regulated by AR. SSH, reverse Northern analysis, and Northern analysis indicated that at doses several orders of magnitude lower than those used for treatment in CHF vesnarinone significantly down-regulated 2 genes (thrombospondin 1, annexin II) up-regulated by AR. The study confirmed earlier findings of AR-mediated alteration in expression of genes that code for noncollagen extracellular matrix (ECM) proteins. Thus, in CF conditioned by exposure to AR, vesnarinone at relatively low doses suppresses genes coding for 2 noncollagen ECM proteins up-regulated by AR. These pharmacologic effects may underlie potentially therapeutic mitigation of fibrosis by vesnarinone. PMID- 15356429 TI - Analysis of US Food and Drug Administration review intervals for drugs approved during the period 1997-2002. AB - This research investigates US Food and Drug Administration review intervals in the United States from 1997 through 2002 for three main categories new drugs applications: new drug application (NDA), the supplemental new drug application (SNDA), and the abbreviated new drug application (ANDA). Review interval for each application was the basis for evaluation and was calculated based on the difference between approval date and the application date. The median review time for all applications was 13.5 months (1.1 years). The median review intervals for ANDAs, NDAs, and SNDAs were 19.1, 12.0, and 10.0 months (1.6, 1.0, and 0.8 years), respectively, during the period. It was found that the median review period for an ANDA was significantly longer than that for an NDA and SNDA. Comparison of application class medians revealed significant differences for all 3 pairwise comparisons (all P < 0.001, Tukey HSD). Within each application category, we compared differences between years. The year effect was not statistically significant for ANDAs or NDAs. NDA median review times were 13.7, 12.0, 12.0, 10.8, 12.5, and 11.7 months (1.14, 1.00, 1.00, 0.90, 1.04, and 0.98 years), while ANDA median review times were 20.4, 19.1, 19.9, 18.6, 18.4, and 21.5 months (1.70, 1.59, 1.66, 1.55, 1.53, and 1.79 years) for 1997, 1998, 1999, 2000, 2001, and 2002, respectively. Year differences were significant for SNDAs (P < 0.001). The primary source of this difference was a lower median review time during 1997, but there was little difference in median review times for the remaining years. PMID- 15356430 TI - Urinary tract diseases revealed after DTP vaccination in infants and young children: cytokine irregularities and down-regulation of cytochrome P-450 enzymes induced by the vaccine may uncover latent diseases in genetically predisposed subjects. AB - Prophylactic vaccinations may sometimes shorten the incubation period of some illnesses and/or convert a latent infection/inflammation into a clinically apparent disease. Cytokines play a major role in mediating the inflammatory process in various clinical entities and represent a potential source of tissue damage if their production is not sufficiently well controlled. It seems that irregularities in production of proinflammatory cytokines may be responsible for the abnormalities associated with full-blown clinical symptoms of various urinary tract diseases observed after DTP vaccination in 13 infants and young children hospitalized over the past 24 years. On admission, upper respiratory tract diseases, atopic dermatitis, and/or latent urinary tract infection/inflammation were found in these children. It is suggested that the whole-cell pertussis present in DTP vaccine, acting as an excessive stimulus in these patients, produced symptoms reminiscent of biologic responses to circulating proinflammatory monokines such as IL-1beta, TNF-alpha, and IL-6 because earlier it was reported that in vitro the whole-cell vaccine induced significantly more such cytokine production than did the acellular pertussis or diphtheria-tetanus only vaccine. Analysis of the cellular immune disturbances previously reported in urinary tract infection/inflammation (increased serum and/or urinary IL-1alpha, IL-1 receptor antagonist, IL-6 and IL-8), steroid-sensitive nephrotic syndrome (increased IL-2, IFN-gamma, TNF-alpha, and decreased or increased IL-4, depending on the cells studied), and atopic dermatitis (decreased IFN-gamma and increased IL-4 production), may suggest that similar subclinical chronic cytokine-mediated abnormalities produced in the course of latent diseases revealed in our patients, combined with those caused by DTP vaccination stimulus, were responsible for the pathomechanism of these clinical entities. This speculation is in agreement with the reports on the long-lasting induction of cytokine release and down-regulation of hepatic cytochrome P-450 isoenzyme activities after administration of DTP vaccine to mice and may be supported by the fact that TH1 phenotype is associated with the up-regulation of intercellular adhesion molecule-1 and RANTES, whereas TH2 phenotype is associated with the up-regulation of the vascular cell adhesion molecule and P-selectin, which are key players in the migration into inflamed tissues and localization of lymphocytes and other allergic effector and inflammatory cells. Because several inflammatory cytokines down-regulate gene expression of major cytochrome P-450 and/or other enzymes with the specific effects on mRNA levels, protein expression, and enzyme activity, thus affecting the metabolism of several endogenous lipophilic substances such as steroids, lipid-soluble vitamins, prostaglandins, leukotrienes, thromboxanes, and exogenous substances, their irregularities in the body may eventually lead to the flare of latent diseases in some predisposed subjects. Also, interleukin genetic polymorphisms, especially the constellation of TNF-alpha and IL-6 genetic variants, might predispose some infants with infection to a more than usually intense inflammatory response in the kidneys after vaccination. It seems that the aforementioned pathomechanism may also be responsible for some cases of sudden infant death syndrome, which is often preceded by infection/inflammation. PMID- 15356431 TI - Opioid therapy and immunosuppression: a review. AB - The idea that opioids modulate the immune system is not new. By the late 19th century, Cantacuzene, used morphine to suppress cellular immunity and lower the resistance of guinea pigs to bacterial infection. While exogenous opioids mediate immunosuppression, endogenous opiates exert opposite actions. Acute and chronic opioid administration is known to have inhibitory effects on humoral and cellular immune responses including antibody production, natural killer cell activity, cytokine expression, and phagocytic activity. Opiates behave like cytokines, modulating the immune response by interaction with their receptors in the central nervous system and in the periphery. Potential mechanisms by which central opiates modulate peripheral immune functions may involve both the hypothalamic pituitary-adrenal axis and the autonomic nervous system. The presence of opioid receptors outside the central nervous system is increasingly recognized. Those receptors have been identified not only in peripheral nerves but also in immune inflammatory cells. The immunosuppression mediated by opiates may explain the increased incidence of infection in heroin addicts. Opiates may also promote immunodeficiency virus infection by decreasing the secretion of alpha and beta chemokines (important inhibitory cytokines for the expression of HIV) and at the same time increasing the expression of chemoreceptors CCR5 and CCR3, coreceptors for the virus. The fact that peripheral immunosupression is mediated at least in part by opioid receptors located in the central nervous system and that intrathecally administered opioids do not exert the same immunosuppressive effects may have important clinical implications for those patients receiving long-term opioid therapy for malignant and nonmalignant pain. PMID- 15356432 TI - Prevention of ventricular fibrillation, acute myocardial infarction (myocardial necrosis), heart failure, and mortality by bretylium: is ischemic heart disease primarily adrenergic cardiovascular disease? AB - It is widely, but mistakenly, believed that ischemic heart disease (IsHD) and its complications are the sole and direct result of reduced coronary blood flow by obstructive coronary artery disease (CAD). However, cardiac angina, acute myocardial infarction (AMI), and sudden cardiac death (SCD) occur in 15%-20% of patients with anatomically unobstructed and grossly normal coronaries. Moreover, severe obstructive coronary disease often occurs without associated pathologic myocardiopathy or prior symptoms, ie, unexpected sudden death, silent myocardial infarction, or the insidious appearance of congestive heart failure (CHF). The fact that catecholamines explosively augment oxidative metabolism much more than cardiac work is generally underappreciated. Thus, adrenergic actions alone are likely to be more prone to cause cardiac ischemia than reduced coronary blood flow per se. The autonomic etiology of IsHD raises contradictions to the traditional concept of anatomically obstructive CAD as the lone cause of cardiac ischemia and AMI. Actually, all the signs and symptoms of IsHD reflect autonomic nervous system imbalance, particularly adrenergic hyperactivity, which may by itself cause ischemia as in rest angina. Adrenergic activity causing ischemia signals cardiac pain to pain centers via sympathetic efferent pathways and tend to induce arrhythmogenic and necrotizing ischemic actions on the cardiovascular system. This may result in ischemia induced metabolic myocardiopathy not unlike that caused by anatomic or spasmogenic coronary obstruction. The clinical study and review presented herein suggest that adrenergic hyperactivity alone without CAD can be a primary cause of IsHD. Thus, adrenergic heart disease (AdHD), or actually adrenergic cardiovascular heart disease (ACVHD), appears to be a distinct entity, most commonly but not necessarily occurring in parallel with CAD. CAD certainly contributes to vulnerability as well as the progression of IsHD. This vicious cycle, which explains the frequent parallel occurrence of arteriosclerosis and IHD, an association that appears to be linked by the same cause, comprises a common vulnerability to deleterious adrenergic actions on the myocardium, lipid metabolism, and vascular system alike, rather than viewing CAD and IsHD as having a putative cause and effect relationship as commonly thought. Adrenergic actions can also cause the abnormal lipid metabolism that is associated with CAD and IsHD by catecholamine-induced metabolic actions on lipid mobilization by activation of phospholipases. This may also be part of toxic catecholamine hypermetabolic actions by enhancing deleterious cholesterol and lipid actions in damaging coronary vessels by plaque formation as well as inducing obstructive coronary spasm and platelet aggregation. This may also cause direct toxic necrosis on the myocardium as well as atherosclerosis in blood vessels. In fact, drugs that inhibit adrenergic actions like propranolol, reserpine, and guanethidine all inhibit arteriosclerosis induced by hypercholesterolemia in experimental animals and prevent carotid vascular disease (associated with stroke) in humans. The concomitant development of myocardiopathy and coronary vascular lesions or coronary and carotid artery intimal medial thickening by catecholamine toxicity is reflected by the frequent primary presentation of patients with catecholamine-secreting pheochromocytoma with cardiovascular disease, ie, hypertension arrhythmias, AMI, SCD, CHF, and vascular disease, which represents a clear example of the primary deleterious impact of catecholamines on the entire cardiovascular system causing adrenergic cardiovascular disease. Thus, like myocardiopathy, CAD and atherosclerosis in general may be the consequences of or a complication of catecholamine actions rather than its putative cause. This report shows how prophylactic bretylium not only prevents arrhythmias but prevents myocardial necrosis, shock, CHF, maintains or restores normal contractility, and lowers mortality in AMI patients by inducing adrenergic blockade. PMID- 15356433 TI - Daily needle aspiration versus surgical lavage for the treatment of bacterial septic arthritis in adults. AB - Septic arthritis is a substantial public health problem, accounting for 0.2-0.7% of hospital admissions. However, despite the availability of effective antibiotics, the appropriate approach to adjunctive therapy remains controversial. Although early drainage is essential to minimize the risks of permanent loss of articular function, it is unclear whether the optimal approach involves arthroscopic lavage or daily arthrocentesis; surgeons appear to prefer surgical lavage because their training routinely considers septic arthritis to be a closed-space infection comparable to an abscess, whereas rheumatologists appear to prefer daily arthrocentesis because of its ease and non-invasive nature. There is a paucity of prospective data comparing the two approaches, and the literature is largely retrospective. Herein, we review the available literature concerning drainage of bacterially infected joints, and conclude that in the absence of prospective randomized clinical trials, the best evidence suggests that there is no compelling reason to recommend surgical lavage for the initial management of uncomplicated septic arthritis. PMID- 15356434 TI - [Environment and health: a social expectation, a scientific challenge]. PMID- 15356435 TI - [Dioxin emissions and soft-tissue sarcoma: results of a population-based case control study]. AB - BACKGROUND: In 1998, the French Ministry of Environment revealed that of 71 French municipal solid waste incinerators processing more than 6 metric tons of material per hour, dioxin emission from 15 of them was above the 10 ng international toxic equivalency factor/m3 (including Besancon, emitting 16.3 ng international toxic equivalency factor/m3) which is substantially higher than the 0.1 international toxic equivalency factor/m3 prescribed by a European directive of 1994. In 2000, a macrospatial epidemiological study undertaken in the administrative district of Doubs, identified two significant clusters of soft tissue sarcoma and non Hodgkin lymphoma in the vicinity of the municipal solid waste incinerator of Besancon. This microspatial study (at the Besancon city scale), was designed to test the association between the exposure to dioxins emitted by the municipal solid waste incinerator of Besancon and the risk of soft tissue sarcoma. METHODS: Ground-level concentrations of dioxin were modeled with a dispersion model (Air Pollution Control 3 software). Four increasing zones of exposure were defined. For each case of soft tissue sarcoma, ten controls were randomly selected from the 1990 census database and matched for gender and age. A geographic information system allowed the attribution of a dioxin concentration category to cases and controls, according to their place of residence. RESULTS: Thirty-seven cases of soft tissue sarcoma were identified by the Doubs cancer registry between 1980 and 1995, corresponding to a standardized incidence (French population) of 2.44 per 100,000 inhabitants. Compared with the least exposed zone, the risk of developing a soft tissue sarcoma was not significantly increased for people living in the more exposed zones. CONCLUSION: Before definitely concluding that there is no relationship between the exposure to dioxin released by a solid waste incinerator and soft tissue sarcoma, a nationwide investigation based on other registries should be conducted. PMID- 15356436 TI - Epidemiology of male reproductive function: a field searching for tools. AB - BACKGROUND: An increase in the frequency of disorders of the male human reproductive organs has been described over recent decades. Neither its causes nor its consequences on fecundity, the ability of the couples to produce a live child, are clearly known. This lack of knowledge may partly be due to methodological difficulties specific to reproductive epidemiology. METHODS: We discuss the relevance and limits of some markers of the male aspects of human reproduction, focusing on semen parameters and fecundability, a measure of the probability of pregnancy. RESULTS: Semen parameters are associated with fecundability, although they have a relatively low sensitivity to detect couples with low fecundability. The study of semen parameters proved central to describe the influence of environmental factors on the male side of reproductive function. The main limitation of semen studies is low participation rates and the possible selection biases ensuing. Fecundability can be estimated by collecting waiting time to pregnancy. Its assessment in retrospective studies often excludes the least fecund, those couples remaining childless, which entails a bias and a decrease in statistical power. The prospective approach and an approach relying on the enrollment of a cross-sectional sample of the couples currently trying to obtain a pregnancy (current duration approach) do not have these limitations. Although it has never been used, the assessment of fecundability using the current duration approach is promising both for aetiologic research and monitoring. CONCLUSION: Most of the potential markers of male reproductive function are not assessed in the general population of France, with the exception of the incidence rate of testis cancer, which is currently increasing. We present some alternatives for a monitoring system of reproductive function. PMID- 15356437 TI - [Assessment of management training for low-level community health workers providing care for children with acute respiratory infections in four districts of Senegal]. AB - BACKGROUND: Acute respiratory infections are the leading cause of death in childhood and most child deaths in Senegal occur at home without qualified health care. Despite this situation, only qualified healthcare workers are authorized to prescribe antibiotics. A competency-based training program was developed to improve and assess management of acute respiratory infections in young children aged between 2 and 59 months by low-level educated community health workers (CHWs) in four districts of Senegal. METHODS: In accordance with the strategy developed by the World Health Organisation, educated low-level community health workers in four district of Senegal were given a three-day course on the management of acute respiratory failure. We assessed the effects of the course by comparing pre-training and post-training skills with the Students t test. RESULTS: The results showed that the educated low-level community health workers were capable of acquiring the skills needed to effectively manage children with acute respiratory failure. CONCLUSION: Further evaluation is needed to determine the mid- and long-term effects of the course and supervised post-training activities. PMID- 15356438 TI - [Factors explaining quality of primary health care in Logone Occidental (Chad)]. AB - The extent of the deficit in human and financial resources associated with alarming health indicators in the developing countries has prompted us to undertake an evaluation of the quality of the primary health care provided in our country. The study was designed to identify the determinants of the quality of primary care delivered in the health centers of Western Logone in Chad by comparing systematic data collected from public and private health structures and an adapted questionnaire. Data were collected from 24 healthcare centers providing care for 395699 people, 73.1% of the population of the district of Logone Occidental. The average costs born by patients receiving care delivered by the different centers was not statistically different between the centers financed by public funds and those with denominational support: 944 Fcfa versus 1315 Fcfa. Variable statistically correlated with quality of care were: number of qualified healthcare workers in the center, training level of the center's manager, average patient-born cost of consultation, average healthcare expenditure per capita in the recruitment area (p<0.01). Considering the importance of the medical district in the organization of healthcare systems in the countries of Sub-Saharan Africa, quality assessment must become a continuous, supervised activity enabling proper description of the processes involved and propositions for local solutions designed to improve the quality of care. PMID- 15356439 TI - [Social inequalities in the demand for dental care]. AB - BACKGROUND: Despite excellent general health indices, Quebec is in a difficult situation concerning oral health: tooth loss remains at a high level in adults and reveals important social inequalities. The objective of this research was to show that dental health inequalities reflect inequalities in the demand for dental care. METHODS: For the Dental Health Survey of Quebec 1998-1999, 9930 parents of children aged 5 to 8 years were randomly selected across Quebec and received a questionnaire at their home on the demand for dental care. Among them, 8430 adults completed and sent back the questionnaire (responses rate: 85%). After excluding edentulous persons, the sample was reduced to 6585 parents aged 30 to 44 years. RESULTS: The majority of respondents (76.4% of women and 72.8% of men) visit the dentist in a preventive manner rather than wait until dental problems occur. However, our study shows important disparities: the proportion of preventive attenders increases as income increases. A multiple logistic regression model suggests that there are financial as well as cultural barriers in dental care access. CONCLUSION: The proportion of preventive attenders is high in Quebec and allows practitioners to adopt a preventive management of dental caries. Social disparities are high however, and are associated with financial as well as cultural barriers that need to be reduced. PMID- 15356440 TI - [Assessment of exposure to traffic pollution in epidemiological studies: a review]. AB - BACKGROUND: Automobile exhaust is a major source of air pollution in urban areas. To study health effects of traffic exhaust fumes epidemiologists need specific tools in order to achieve a precise assessment of human exposure to traffic air pollution (TAP) and avoid misclassification. The aim of this review is to study the different ways of assessing human exposure to TAP in epidemiological studies dealing with short-term or long-term health effects of TAP. METHODS: After presenting the different designs and goals of the studies mentioned above, this review focuses on methods of assessing exposure to TAP and their different associated health endpoints. RESULTS: To assess exposure to TAP, most published studies have used more or less complex exposure indices. Several teams have used residence location and its proximity to traffic, traffic counts, or a combination of both. More recently, some authors have developed mathematical dispersion models and statistical regression models. DISCUSSION: Our analysis shows that reliable and validated tools would be needed to assess accurately human exposure to TAP. This can only be achieved with statistical regression models and mathematical dispersion models. Although such methods may be difficult to implement, their use can be facilitated by adding a geographic information system. PMID- 15356441 TI - [Ethical and regulatory aspects of research in clinical epidemiology. Practical guidelines]. PMID- 15356442 TI - The anatomic and physiologic basis of local, referred and radiating lumbosacral pain syndromes related to disease of the spine. AB - Conscious perception and unconscious effects originating from the vertebral column and its neural structures, although complex, have definite pathways represented in a network of peripheral and central nervous system (CNS) ramifications. These neural relationships consequently result in superimposed focal and diffuse, local and remote conscious perceptions and unconscious effects. Any one or combination of somatic and autonomic signs and symptoms may potentially be observed in a particular patient. This variety and inconsistency may mislead or confuse both the patient and the physician. A clear understanding of the basic anatomic and physiologic concepts underlying this complexity should accompany clinical considerations of the potential significance of spondylogenic and neurogenic syndromes in any disease process affecting the spine. PMID- 15356443 TI - Treatment of herniated lumbar disc by intradiscal and intraforaminal oxygen-ozone (O2-O3) injection. AB - MATERIAL: We report our experience between May 1996 and May 2003 with 2200 patients affected by low back pain or sciatica due to herniated disk treated by intradiscal and intraforaminal oxygen-ozone injection. The patients received medical and physical therapy before treatment for at least 2 months; the patients with conus-cauda syndrome and hyperalgesic sciatica were excluded. We never performed discography before the treatment that was performed under CT guidance or fluoroscopy. CT provided monitoring of gas distribution in the disk and epidural space. RESULTS: No side effects were recorded at short and long-term follow-up. Clinical results were evaluated with the modified McNab method showing an 80% success rate and 20% failure rate in 1750 patients followed up to 6 months while the success rate dropped down at 75% and failure increased at 25% in 1400 followed up to 18 months. CT showed reduction in the size of the herniated disk in only 63% of the followed patients (420 patients). The failure has been mostly related to: calcified herniated disk; spinal canal stenosis; recurrent herniated disk with epidural fibrosis; small descending herniated disk at the level of the lateral recess. PMID- 15356444 TI - Estimation of relative blood volume in head and neck squamous cell carcinomas. AB - PURPOSE: MR based first-pass method can be utilized to obtain hemodynamic information in the head and neck region. The purpose of this study was to estimate the regional relative blood volume (rBV) in head and neck tumors, which is useful for tumor staging and tumor biopsy. METHODS: Eighteen patients with head and neck tumors (17 squamous cell carcinomas, 1 hemangiopericytoma) were studied on a 1.5-T system. Conventional T1-weighted MR images and T2-weighted images and sequential T2*-weighted images were obtained. During repetitive image sequence acquisition, a bolus (0.2 mmol/kg) of gadopentetate dimeglumine was mechanically injected. Image processing of the dynamic raw data was performed on a pixel-by-pixel basis. RESULTS: Regional relative blood volume maps of the head and neck were successfully reconstructed in all (18/18) patients. The regional relative blood volume values within the tumor area of squamous cell carcinoma were 7.0 +/- 2.8, normalized on muscle, whereas the rBV of a single hemangiopericytoma was 11.6. The difference of rBV values of tumor and muscle was highly significant at statistical evaluation (p < 0.001). CONCLUSIONS: Relative blood volume imaging of head and neck tumors is valid using MR-based first-pass method. This method provides hemodynamic information which is not available from conventional MR imaging and is promising for further characterization of head and neck tumors PMID- 15356445 TI - [MRI of the brachial plexus]. AB - Magnetic resonance imaging is the method of choice for the evaluation of brachial plexopathy. Knowledge of the anatomy and normal imaging appearance is required. High-resolution imaging technique is necessary with the use of adequate coils. Evaluation of the brachial plexus requires T1 weighted sequences in three plans, T2 weighted sequences with fat suppression and if necessary the study is completed with gadolinium injection sequences with fat suppression. A CISS sequence is used if a nerve root avulsion is suspected. The spatial resolution must be optimized with the use of adapted parameters. We illustrate a variety of pathologies that can involve the brachial plexus. The pathology includes trauma, primary (neurogenic tumors, lymphomatosis) or secondary tumors, radiation plexopathy or inflammatory polyneuropathy. PMID- 15356446 TI - Imaging features of spinal epidural cavernous malformations. AB - Cavernous angioma or cavernoma is a vascular malformation that may affect any area in the neuraxis. Epidural location is very rare and therefore seldom considered in the differential diagnosis of spinal cord compression. We report two cases of epidural cavernous angiomas. The first case is a solitary and purely epidural dorsal cavernous angioma without foraminal expansion or bone modification causing spinal cord compression in a 35 year old woman. The second case is a solitary epidural dorsal cavernous angioma with foraminal extension causing spinal cord compression in a 56 year old woman. Histological confirmation is available for both cases. We describe the MRI features of this lesion insisting on its differential diagnosis on imaging. PMID- 15356447 TI - [Spinal extradural meningiomas: MRI findings in two cases]. AB - Spinal extradural meningiomas are rare and may be easily confused with malignant neoplasms. We report two unusual cases of epidural spinal meningioma one within the left C6-C7 foramen and the other within the left posterolateral epidural space at the T3-T4 level. Low signal intensity of the tumor on T2-wi, thickening and enhancement of the dura with only the possibility of bone erosion are the most characteristic MR findings. PMID- 15356448 TI - [Cavernous sinus dural arteriovenous fistula complicated by edematous cerebral lesions from venous etiology]. AB - The authors report the case of a 38-year-old woman with a cavernous DAVF resulting in edematous lesions located in the territory of the ipsilateral basal vein. Transarterial embolization led to subtotal regression of the fistula associated with the regression of cerebral abnormalities. The authors discuss the pathophysiology of the cerebral edematous lesions and the therapeutic consequences according to the venous drainage of the cavernous sinus. PMID- 15356449 TI - Epidermoid tumor of the pons. AB - Epidermoid tumors originating from the brainstem are extremely rare. The authors report a patient with an intraaxial epidermoid tumor of the pons. The tumor involved most of the pons and had a small exophytic component. PMID- 15356450 TI - Facial infantile hemangiopericytoma resembling an arteriovenous malformation. AB - Malignant highly vascularized tumors such as hemangiopericytomas (HPC) may mimic a benign arteriovenous malformation (AVM) which is sometimes still referred to as "angioma". We describe the clinical and radiological findings of a facial hemangiopericytoma in comparison to an AVM in order to avoid misdiagnosis between these two pathologies since evolution and therapeutic management are completely different. Because hemangiopericytomas in children show malignant behavior requiring aggressive management, early and accurate diagnosis is of significant importance for the clinical outcome. PMID- 15356451 TI - Transient tic disorder following carbon monoxide poisoning. AB - We report a 12-year-old male patient who developed transient motor and vocal tics twelve days after carbon monoxide (CO) poisoning. Cranial magnetic resonance image (MRI) of the patient showed bilateral symmetric hyperintensity in the caudate nucleus and putamen. Tic disorder was successfully treated with haloperidol. Thirty-three months after CO poisoning, the patient was asymptomatic and MRI revealed atrophy in caudate nucleus and putamen. The mechanism of tic disorder in CO intoxication is discussed. PMID- 15356452 TI - Diffusion-weighted MR imaging and pathologic findings in adult cerebellar medulloblastoma. AB - OBJECTIVES, MATERIALS AND METHODS: The authors present the diffusion-weighted MR imaging and pathologic findings in two adult patients with cerebellar medulloblastoma. RESULTS: Both presented with a vermian mass of the posterior fossa with low signal on SE T1 weighted images, and moderate enhancement of the mass after gadolinium injection. The tumors were of high intensity on diffusion weighted images with low ADC value. The ADC values (x10(-3) mm2/s) were respectively 0.60 +/- 0.06 and 0.59+/-0.11 (tumor), and 0.65 +/- 0.04 and 0.67 +/ 0.07 (cerebellar white matter). Tumors were highly cellular and composed of densely packed small round cells with hyperchromatic nuclei and scanty cytoplasm. CONCLUSION: diffusion-weighted MR imaging may be useful for the diagnosis of cerebellar medulloblastoma, due to their high cellularity and high nuclear-to cytoplasmic ratio. PMID- 15356453 TI - Lowe syndrome: proton mr spectroscopy, and diffusion mr imaging. AB - A 3-year-old boy with Lowe syndrome had bilateral periatrial hyperintense lesions intermixed with small cyst-like changes in the periatial regions of the deep white matter at MR imaging. Proton MR spectroscopy revealed prominent myoinositol peaks suggesting the presence of gliosis. b = 1000 s/mm2 images of diffusion MR imaging were negative for the periatrial lesions. ADC maps, however, revealed high ADC values (1.76, and 1.66 x 10(-3) mm2/s), compared to the normal brain parenchyma (0.81 x 10(-3) mm2/s). These diffusion MR imaging findings likely represented gliosis. PMID- 15356454 TI - Physical health status of World Trade Center rescue and recovery workers and volunteers - New York City, July 2002-August 2004. PMID- 15356455 TI - Mental health status of World Trade Center rescue and recovery workers and volunteers - New York City, July 2002-August 2004. AB - After the September 11, 2001, attacks on the World Trade Center (WTC), a comprehensive screening program was established to evaluate the physical and mental health of rescue and recovery workers and volunteers. Persons were eligible for this program if they participated in the WTC rescue or recovery efforts and met specific time criteria for exposure to the site. During July 16, 2002--August 6, 2004, the program evaluated 11,768 workers and volunteers. This report summarizes data analyzed from a subset of 1,138 of the 11,768 participants evaluated at the Mount Sinai School of Medicine during July 16--December 31, 2002. On the basis of one or more standardized screening questionnaires, approximately half (51%) of participants met threshold criteria for a clinical mental health evaluation. Continued surveillance is needed to assess the long term psychological impact of the aftermath of the 9/11 attacks and to determine needs for continued treatment. PMID- 15356456 TI - Preliminary results from the World Trade Center Evacuation Study - New York City, 2003. PMID- 15356457 TI - Investigation of acute idiopathic pulmonary hemorrhage among infants - Massachusetts, December 2002-June 2003. PMID- 15356458 TI - Progress towards poliomyelitis eradication - Egypt, 2003-2004. PMID- 15356459 TI - West Nile virus activity - United States, September 1-7, 2004. PMID- 15356460 TI - Intravenous bisphosphonates and osteonecrosis. PMID- 15356461 TI - Clinicopathologic conference: trismus following dental treatment. AB - Myositis ossificans traumatica is a rare clinical entity in the maxillofacial region. We present a case of myositis ossificans traumatica of the temporalis muscle following dental treatment in the form of a clinicopathologic conference. A review of the literature supporting our findings is provided. PMID- 15356462 TI - Tuberculosis: medical management update. AB - Tuberculosis (TB) is an infectious chronic disease. After decades of steadily declining prevalence, the disease has reemerged in the last 5 years. Symptoms of TB are mild and not specific and can be classified as either systemic or localized to target organs. Microscopic examination of the sputum remains an inexpensive and rapid way to identify highly infectious patients. Four different antimicrobial agents-rifampin, ethambutol, pirazinamide, and isoniazid-form the basis of currently recommended antituberculosis therapy. Tuberculosis could be an occupational risk for health care workers. Dentists must be involved in the health promotion and early detection of TB. PMID- 15356463 TI - Coronectomy (intentional partial odontectomy of lower third molars). AB - OBJECTIVE: This study evaluates the rate of infection of retained lower third molar roots after coronectomy in teeth judged to be in intimate relation to the inferior alveolar nerve. STUDY DESIGN: This was a retrospective study of 52 patients who were operated on over a 10-year period. RESULTS: Only 3 of 52 patients had to have roots removed because of pain or infection. Postoperative findings and aspects of the surgical technique are also discussed. CONCLUSIONS: Coronectomy is a worthwhile option to extraction where a lower third molar is judged to be in close proximity to the inferior alveolar nerve. In lower third molar removal the potential damage is high when certain radiographic signs are present, whereas the infection rate of roots remaining after coronectomy is, by contrast, low. PMID- 15356464 TI - Clinical review of 580 ranulas. AB - OBJECTIVE: The purpose of this paper was to compare clinical features among 3 patterns of ranula and the recurrence rates of each when treated by different surgical methods. METHODS: A retrospective review of clinical and pathologic records in 580 ranulas was undertaken. Ranulas were classified into 3 clinical types according to sites of the primary swelling: oral ranula, plunging ranula, and mixed ranula. Information was collected on age at presentation, sex, history of onset, sites of swelling, surgical methods, histological findings, and outcome of treatment. RESULTS: Ranula was most prevalent in the second decade of life and slightly more common in females (male to female ratio of 1:1.2), but a distinct male predilection was noted for the plunging ranula (male to female ratio of 1:0.74). Oral ranula was most commonly involved in the left side (left to right ratio of 1:0.62), while the plunging and mixed ranula were commonly involved in the right side (left to right ratio of 1:1.38, 1:1.16 respectively). In the plunging ranula group, there were more patients who had the history more than 6 months. The recurrence rates of ranulas were not related to swelling patterns and surgical approaches, but intimately related to the methods of surgical procedures. The recurrent rates for marsupialization, excision of ranula, and excision of the sublingual gland or gland combined with lesion were 66.67%, 57.69%, and 1.20%, respectively. CONCLUSION: Three patterns of ranula have similar clinical and histopathologic findings, although plunging ranula has some different clinical features. Removal of the sublingual gland via an intraoral approach is necessary in the management of various clinical patterns of the ranula. Recurrence rates of ranulas of any type are excessive unless the involved sublingual gland is removed. PMID- 15356465 TI - Effectiveness of an educational program in reducing the incidence of wrong-site tooth extraction. AB - OBJECTIVES: The aim of the study was to investigate the effectiveness of an educational program on the reduction of the incidence of wrong-site tooth extraction at the outpatient department of a university hospital in Taiwan. STUDY DESIGN: Data collected from cases of wrong-site tooth extraction during 1996 to 1998 were used to develop a specific educational intervention that was implemented from 1999 to 2001. The annual incidence of erroneous extraction was compared between the preintervention and intervention periods. The factors contributing to wrong tooth extraction were also analyzed. RESULTS: The annual incidence rates of erroneous extraction from 1996 to 1998 were 0.026%, 0.025%, and 0.046%, respectively. During the intervention period from 1999 to 2001, wrong site tooth extraction did not occur at the department. There was a significant difference in the incidence of erroneous extraction between the preintervention and intervention periods (P<.01). Cognitive failure was the most frequent form of active failure responsible for wrong-site tooth extraction, whereas communication and training were found to be major latent factors contributing to these errors. CONCLUSIONS: Our results suggest the effectiveness of an educational program comprising case-based materials, information feedback, and clinical guidelines in reducing the incidence of wrong-site tooth extraction. PMID- 15356466 TI - Central odontogenic fibroma of mandible: a case report and review of the literature. AB - Central odontogenic fibroma is an extremely rare benign tumor, accounting for less than 0.1% of all odontogenic tumors. The most recent literature review revealed that only 69 cases have so far been reported in the English literature, and only 1 of these cases showed radiologic appearance of a dentigerous cyst and this involved impacted maxillary incisors. A case of central odontogenic fibroma occurring in the mandible of a 30-year old female with radiological appearance of a dentigerous cyst involving an impacted lower first molar, presented here, appears to be the first reported case of central odontogenic fibroma of mandible with radiological appearance of dentigerous cyst in the English-language literature. PMID- 15356467 TI - Oropharyngeal candidiasis in HIV-infected patients under treatment with protease inhibitors. AB - OBJECTIVE: Oropharyngeal candidiasis decreased when protease inhibitors were included with other antiretrovirals to treat HIV infection. We tested oral yeast isolates of Brazilian HIV-infected individuals receiving antiretroviral therapy for protease secretion and susceptibility to ritonavir and some antifungals. STUDY DESIGN: We collected oral samples and identified yeasts from 19 HIV infected patients receiving highly active antiretroviral therapy (HAART) and suspected of having oral candidiasis. Ritonavir and its excipients' effects on the isolated yeasts were tested for protease secretion by Ruchel's technique. The yeasts' susceptibility to amphotericin B (AnB), fluorocitosine (5FC), fluconazole (FZL), ketoconazole (KZL), and itraconazole (IZL) was determined by E-test (AB Biodisk). Chi-squared test determined the statistical differences. RESULTS: Twenty-five different positive isolates were obtained. Sixty-eight percent were C. albicans. Other isolates included C. famata (16%), C. glabrata (4%), C. tropicalis (4%), T. capitatum (4%), and 1 isolate not identified. High protease secretion was observed for most of the isolates (20/25). Ritonavir only altered enzyme secretion in 6/20 of the protease-secreting isolates. All isolates were highly sensitive to both AnB and 5FC. Antifungal activity did not change when ritonavir was added to the culture media. Some isolates were highly resistant to studied antifungals (52.2% KZL, 30.4% FZL, and 26% IZL). Resistance significantly decreased when ritonavir was added to the medium with KZL and IZL (P <.5 by chi squared). A trend to decreased resistance was also observed with FZL but the results were not statistically significant. CONCLUSION: Candida continues to be the most prevalent fungus in the oral cavity. Although oral candidal isolates secrete protease, ritonavir does not inhibit all protease-secreting oral yeast isolates. There seems to be a synergistic effect between ritonavir and oral antifungals against fungal resistance. PMID- 15356468 TI - Idiopathic trigeminal neuralgia: clinical aspects and dental procedures. AB - OBJECTIVE: Evaluation of the characteristics and dental treatments in a Brazilian series of idiopathic trigeminal neuralgia (ITN) patients. STUDY DESIGN: Forty eight subjects with ITN were interviewed and evaluated according to a systematized approach. RESULTS: FINDINGS: (a) location of the trigger zone, intraoral in 64.58%; (b) comorbidities, 18 patients (hypertension in 9); (c) treatment with dental procedures for ITN according to duration of pain (31), 44.44% (during the first year), 50.00% (1-5 years), 80.00% (5-10 years), and 100.00% (>10 years). There was no correlation between trigger zone location and the frequency of previous dental procedures (P=.667). There was significant statistical correlation between the number of patients who underwent dental treatments and the duration of ITN (P=.004). CONCLUSION: (a) Demographic characteristics of this sample are similar to those described in the literature, (b) the dental procedures were not correlated with location of trigger zone, and (c) patients with long-lasting ITN had more number of previous dental procedures. PMID- 15356469 TI - A case of neurofibromatosis-Noonan syndrome with a central giant cell granuloma. AB - Neurofibromatosis Type 1 (NF1) is one of the most common autosomal dominant diseases affecting multiple systems including the vascular, skeletal, and central nervous system. Noonan syndrome (NS) is an autosomal dominant genetic disorder, associated with musculoskeletal and skin manifestations. Coexistence of central giant cell lesions in patients with both NS and NF1 were reported in the literature. Development of multiple central giant cell lesions in a patient with a Noonan syndrome has been referred to as Noonan-like syndrome. A few cases with features of NF1 and NS have been termed as NF1-NS. Here, we present a case of so called NF-NS associated with a central giant cell lesion. PMID- 15356470 TI - Fusospirochetosis causing necrotic oral ulcers in patients with HIV infection. AB - Under certain permissive circumstances, normally occurring fusiform bacteria and Borrelia spirochetes can result in a symbiotic overgrowth that leads to necrotic oral ulcers (stomatitis), gingivitis, and periodontitis. These lesions are collectively known as oral fusospirochetosis and may be under-appreciated in patients with HIV infection and AIDS. Fusospirochetal oral ulcers in patients with HIV are often large, necrotic, and malodorous; they respond completely to penicillin. We report 3 patients with HIV infection and fusospirochetal ulcerative stomatitis and review the clinical presentation, microbiologic diagnosis, potential pathogenesis, and treatment of these lesions. PMID- 15356471 TI - Basic fibroblast growth factor (bFGF) in saliva and oral mucosa in patients with oral lichen planus: preliminary observations. AB - OBJECTIVE: Basic fibroblast growth factor (bFGF) is important for wound healing and tissue repair. This study measures the concentration of bFGF in oral lichen planus (OLP) affected mucosa and in the saliva of patients with OLP. STUDY DESIGN: Samples of saliva, OLP-affected mucosa, and clinically healthy mucosa were obtained from 11 patients. Control samples were obtained from healthy volunteers. The bFGF content of tissue samples and saliva was examined by ELISA. RESULTS: The mean bFGF concentration in saliva from OLP patients was 5.9 pg/mL, SD 2.9, compared with 0.3 pg/mL, SD 0.3, in the control group, (P>.01). The bFGF content in the OLP tissue was 90.6 microg/mg protein, SD 39.5, in clinically normal mucosa from OLP individuals it was 46.2 microg/mg protein, SD 12.0 (P=.02), and in the control group 46.2 microg/mg protein, SD 11.5 (P>.01). CONCLUSION: OLP-affected mucosa contained significant more bFGF than nonaffected mucosa in OLP and healthy mucosa in control group. There is no difference between nonaffected mucosa in OLP and control group. Saliva in OLP patients contained more bFGF than saliva in control patients. PMID- 15356472 TI - Glomus tumor: a rare location in the upper lip. AB - Glomus tumor is a benign neoplasm composed of glomocytes and vessels in varying proportions. It appears as a single painful nodule typically found in the dermis and the subcutaneous tissue of the extremities. A rare appearance of the tumor in the upper lip is presented. Differential diagnosis (along with clinical, microscopic, and immunohistochemical findings) is discussed. PMID- 15356473 TI - Ultrasonographic features of the masseter muscle in female patients with temporomandibular disorder associated with myofascial pain. AB - OBJECTIVE: The purpose of the present study was to examine ultrasonographic appearances in female patients with temporomandibular disorder (TMD) associated with myofascial pain in comparison with healthy volunteers. STUDY DESIGN: The thickness of the masseter muscle in 25 female patients with TMD was measured at rest and at maximum contraction using ultrasonography. The visibility and width of the internal echogenic bands of the masseter muscle were also assessed and the muscle appearance was classified as 1 of 3 types: type I, characterized by the clear visibility of the fine bands; type II, thickening and weakened echo intensity of the bands; type III, disappearance or reduction in number of the bands. RESULTS: There were significant differences in the thickness at rest and the increase ratio by contraction between the patient and control groups. The distribution of muscle types showed a significant difference between the 2 groups. CONCLUSION: The ultrasonographic features of the masseter muscle in TMD patients with myofascial pain were clarified and they might be related to muscle edema. PMID- 15356474 TI - Contributions of reformatted computed tomography and panoramic radiography in the localization of third molars relative to the maxillary sinus. AB - OBJECTIVE: The aim of this clinical study was to seek an answer to the question of whether CT gives better qualitative and quantitative information as compared to panoramic radiograph when deciding on extraction of upper impacted third molars. STUDY: The study concerned 34 third molars appearing in direct relation with the sinus on the panoramic radiograph for which a CT scan had been recommended. For each case, the panoramic radiograph was compared with the CT scan for 5 evaluation criteria. RESULTS: The CT scan was more precise than the panoramic radiograph by 1.67 mm for measurement of the level of impaction of the third molar, by 12.44 degrees for measurement of the third molar displacement, by 0.74 mm for bone height separating the third molar roots and the sinus, and by 2.26 mm for the length of roots in the sinus. The CT scan was equally accurate for measurement of the distance separating the crowns of second and third molars in 34 cases. CONCLUSIONS: This study confirms the gain in precision for the CT scan as compared with conventional techniques. The CT scan therefore presents advantages when considering the extraction of upper third molars by providing both qualitative and quantitative information liable to bear on the diagnosis and therapeutic management. PMID- 15356475 TI - Alternation of the magnetic resonance signals characteristic of mandibular condyles during growth. AB - OBJECTIVE: In a previous study, we named the magnetic resonance imaging (MRI) signal that is characteristic of mandibular condyles in growing children the "double contour-like structure" (DCLS). In the current study, we evaluated the alternating appearance and disappearance of the DCLS in children as they grow to elucidate the significance of the DCLS. STUDY DESIGN: In 11 subjects who were 9 14 years old at the time of the first examination, MRI characteristics of mandibular condyles were sought from fast spin-echo intermediate-weighted images; 2 or 3 years later, repeat MRI were obtained from the same subjects. RESULTS: At the second examination, 10 of the 16 DCLSs detected at the first examination had disappeared. A significant relationship was found between the alternation of the DCLS and the bone marrow conversion from red to yellow in mandibular condyles. Alternation of the DCLS tended to coincide with eruption of the permanent maxillary second molars. The DCLS was also shown to appear as a hypersignal on T2 weighted images. CONCLUSIONS: Double contour-like structure signals in mandibular condyles appeared and disappeared as children grew, and these signal alternations may be useful as a criterion for the staging of mandibular condyle development in children. Possible constituents of the DCLS may be hypervascular loose fibrous tissue and a proliferation of cartilage. PMID- 15356476 TI - MR study of lateral retropharyngeal lymph nodes at different ages. AB - OBJECTIVE: The purpose of this study was to investigate the age-related morphologic changes of the lateral retropharyngeal lymph nodes using MR imaging. STUDY DESIGN: The maximal axial diameter of the nodes was measured on the MR image in 120 healthy subjects (younger group 6-19 years (n = 40), middle group 20 38 years (n = 48), older group 42-74 years (n = 32)). Between-group differences in the diameter of the nodes were analyzed. RESULTS: Mean values of the maximal axial diameter of the nodes were 9.0 +/- 1.6 mm, 6.6 +/- 1.7 mm, and 5.3 +/- 1.6 mm, corresponding to younger group, middle group, and older group, respectively ( P <.01). CONCLUSION: The information obtained in the current study is useful for the differential diagnosis of normal and abnormal lateral retropharyngeal lymph nodes according to age. PMID- 15356477 TI - Antibacterial efficacy of calcium hydroxide, iodine potassium iodide, betadine, and betadine scrub with and without surfactant against E faecalis in vitro. AB - OBJECTIVE: This study investigated the ability of endodontic irrigants and medicaments to eliminate Enterococcus faecalis from infected dentinal tubules, and whether their antimicrobial action was enhanced by surfactant. STUDY DESIGN: For the study, 5-mm dentin disks were sectioned from bovine incisor roots and infected with E faecalis. Lumens were instrumented, and 1 of 7 medicaments (10% Ca(OH) 2, Betadine, or IKI, each with or without surfactant, or Betadine Scrub) was used to flush and fill each lumen. Positive controls received saline. Specimens were incubated for 15 minutes or 24 hours. Quantitative microbiology of the remaining bacteria was performed and groups were compared using a 1-way ANOVA. RESULTS: The addition of surfactant did not enhance the antibacterial action of any medicament. When used as a 24-hour medicament, Ca(OH) 2 consistently failed to eliminate E faecalis, whereas both Betadine Scrub and IKI rendered 90% of samples sterile. IKI was the only agent shown to consistently eliminate E faecalis in a 15-minute time frame. CONCLUSION: Under the in vitro conditions of this study, IKI was able to eliminate E faecalis from bovine root dentin when used with a 15-minute contact time. PMID- 15356478 TI - Papilla healing following sulcular full thickness flap in endodontic surgery. AB - OBJECTIVE: The objective was to analyze recession of the interdental papilla in periodontally healthy situations following apical surgery using sulcular flap incisions. STUDY DESIGN: Apical surgeries were performed in 13 patients with good periodontal health. A sulcular full thickness flap involving at least 1 interproximal space was raised, using microsurgical instruments and magnification with the least possible trauma. The papillae were closed with vertical mattress sutures (polyamide 6/0) and removed after 3-5 days. The height of the interdental papilla was evaluated preoperatively and postoperatively at 1 month and 3 months using plaster casts and measured with a laser scanner. Thirteen surgical sites were compared to untreated papillae. The pairwise t test was used for statistical analysis. RESULTS: All experimental sites exhibited a significant loss of papilla height at 1 month (P <.003) and 3 months (P <.004). At 3 months retractions increased in 10 sites, while in 3 sites the loss had diminished compared to the 1 month value. CONCLUSIONS: These results suggest that the conventional sulcular flap results in considerable retraction of the papilla height after 1 month and 3 months postsurgically. PMID- 15356479 TI - Direct contact between enamel matrix derivative (EMD) and osteoblasts is not required for EMD-induced cell proliferation. AB - OBJECTIVE: The purpose of this study was to determine whether the effect of enamel matrix derivative (EMD) on osteoblast proliferation is dependent on direct contact between EMD and the cells. STUDY DESIGN: MC3T3-E1 cells were seeded onto 6-well culture plates at an initial density of 5000/cm(2) in Dulbecco's modified eagle medium (DMEM) with 10% fetal bovine serum (FBS). Serum was removed from the culture medium after 24 hours with or without the addition of EMD. Four groups were evaluated: group 1, DMEM only; group 2, DMEM with 100 microg/mL EMD directly added to the culture medium; group 3, DMEM with a culture plate insert (30-mm diameter; 0.4-microm pore size) only; group 4, DMEM with 100 microg/mL EMD added onto a culture plate insert. The porous membrane of the insert prevented direct contact between EMD and the cells. After 3-day incubation, cell morphology was examined and the total cell number per well was counted and analyzed using 1-way ANOVA. RESULTS: EMD formed precipitated aggregates on the membrane of the culture insert with the same appearance as when it was added directly to the medium. The culture plate insert alone did not cause any changes in cell morphology or proliferation. The addition of EMD significantly increased cell number regardless the presence of the culture plate insert. CONCLUSION: This study suggests that direct contact between EMD and osteoblasts is not required to induce cell proliferation. Soluble peptides released from EMD may contribute to the stimulating effects of EMD on cell proliferation. PMID- 15356480 TI - Histologic evaluation of pulpotomies in dog using two types of mineral trioxide aggregate and regular and white Portland cements as wound dressings. AB - OBJECTIVE: The purpose of this study was to investigate the pulpal response of dogs' teeth after pulpotomy and direct pulp protection with MTA Angelus, ProRoot, Portland cement and white Portland cement. STUDY DESIGN: Seventy-six teeth were treated with these materials. One hundred twenty days after treatment, the animals were sacrificed and the specimens removed and prepared for histological analysis. RESULTS: All the materials demonstrated similar results when used as pulp-capping materials. Pulp vitality was maintained in all specimens and the pulp had healed with a hard tissue bridge. CONCLUSION: The materials used in this study were equally effective as pulp protection materials following pulpotomy. PMID- 15356481 TI - Advances in Orthodontics and Dentofacial Surgery. Proceedings of the AAO/AAOMS symposium. February 6-8, 2004, Palm Springs, California, USA. PMID- 15356482 TI - Mandibular midline osteotomy for constriction. PMID- 15356483 TI - Practical applications of distraction osteogenesis. PMID- 15356484 TI - Stability and predictability of orthognathic surgery. PMID- 15356485 TI - Surgical and orthodontic management of impacted maxillary canines. PMID- 15356486 TI - Management of missing maxillary anterior teeth with emphasis on autotransplantation. PMID- 15356487 TI - Recovery after third-molar surgery. PMID- 15356488 TI - Facial planning for orthodontists and oral surgeons. PMID- 15356489 TI - Open-bite closure with mandibular osteotomy. PMID- 15356490 TI - Surgical procedures to improve esthetics when orthognathic surgery is not an option. PMID- 15356491 TI - Implant-anchored orthodontics for partially edentulous malocclusions in children and adults. PMID- 15356492 TI - Plastic surgery combined with orthodontic and orthognathic procedures. PMID- 15356493 TI - Three-dimensional craniofacial imaging. PMID- 15356494 TI - The effect of cervical headgear on patients with high or low mandibular plane angles and the "myth" of posterior mandibular rotation. AB - The purpose of this retrospective survey was to compare the magnitude of posterior mandibular rotation during orthodontic treatment with edgewise appliances and cervical headgear in patients with high or low Frankfort mandibular plane angles (FMA). The files of a private orthodontic practice were searched, and the records of growing patients with Class II Division 1 malocclusion were selected. These were divided into 2 groups, according to FMA: the hypodivergent (FMA < 22 degrees ) or low-angle group consisted of 29 patients with a median FMA of 19.80 degrees, and the hyperdivergent (FMA > 28 degrees ) or high-angle group consisted of 31 patients with a median FMA of 32.70 degrees. All patients had nonextraction treatment with full edgewise appliances (Roth prescription), including second molars, by the same clinician. Cervical headgear and Class II elastics were used, and several patients also had fixed or removable anterior biteplanes for short periods. Pretreatment and posttreatment cephalometric tracings were superimposed on internal basic structures of the mandible. There was no difference in FMA changes between the 2 groups, nor were there statistically significant differences in changes during treatment, with 2 exceptions: the angle SN-GoGn showed a very small (0.86 degrees ) mean differential change between groups, generated mostly by the counterclockwise mandibular rotation of the low-angle patients. This was verified with structural superimpositions to evaluate vertical changes. In addition, posterior face height increase was significantly greater in the low-angle group. Structural superimposition of the mandible after treatment showed marked counterclockwise rotations in relation to the anterior base of the skull in both groups, with the high-angle group rotating significantly less. On average, growth and treatment resulted in improvements in the high-angle patients but aggravated the problems in the low-angle patients with deep bite malocclusions. PMID- 15356495 TI - Assessing patient-specific decalcification risk in fixed orthodontic treatment and its impact on prophylactic procedures. AB - This longitudinal and prospective clinical study determined the decalcification rate during multibracket treatment (mean duration, 22 months) in 80 randomized, selected adolescents divided into 4 groups of 20 patients each. The study had 2 objectives. The first was to establish whether patient selection based on predefined caries risk indicators, ie, plaque index, approximal plaque index, D(3 4)MFT/d(3-4)mft index, initial lesions, and gingiva index, carried out before the start of treatment allows for valid predictions of the decalcification risk associated with multibracket treatment. The second objective was to determine whether an extended prophylaxis regimen, including regular mechanical tooth cleaning by a hygienist, scaling, and chlorhexidine treatment, as an adjunct to an initial prophylaxis introductory program consisting of patient motivation, oral hygiene checkups, and fluoridation, reduces or prevents decalcification more effectively than initial prophylaxis alone. The results showed that patient selection based on caries risk factors provides a simple and effective method for predicting the decalcification risk in multibracket treatment (P .05). This new 3-dimensional methodology proved useful for comparing treatment outcomes by evaluating the morphologic changes induced by palatal expansion and generated a better visualization of these outcomes. PMID- 15356501 TI - Clinical and microbiologic changes after removal of orthodontic appliances. AB - The goal of this study was to evaluate the clinical and microbiological factors associated with orthodontic appliances during an episode of gingival inflammation and the impact of appliance removal on periodontal health. This prospective study included 10 patients, aged 12 to 20 years, with clinical signs of gingival inflammation at the final phase of orthodontic treatment (appliance removal). Plaque index, gingival index, and probing depth were evaluated, and microbiological samples were collected from teeth 16, 11, and 26 at 2 times: during the gingival inflammation (baseline) and 30 days after the removal of the appliance and professional prophylaxis. Polymerase chain reaction analysis was used to detect Porphyromonas gingivalis, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Prevotella nigrescens. A statistically significant improvement of the plaque and gingival indexes was seen, as well as a reduction in probing depth (P <.05). Periodontal pathogens were associated with gingival inflammation during orthodontic treatment. The improvement in periodontal health at 30 days was concomitant with a reduction of sites positive for A. actinomycetemcomitans and B. forsythus (P <.05). Periodontal pathogens associated with gingival inflammation during orthodontic treatment can be significantly reduced by orthodontic appliance removal and professional prophylaxis. PMID- 15356502 TI - Salivary flow rates during relaxing, clenching, and chewing-like movement with maxillary occlusal splints. AB - The purpose of this study was to test the hypothesis that the application of occlusal splints increases the diurnal salivary flow rate both in bruxism patients and in normal subjects. Salivary flow rates in 16 adult volunteers (8 bruxism patients and 8 sex- and age-matched control subjects) were measured with the spitting method. There was no significant difference in the salivary flow rate with or without splints between the control and bruxism groups. In all subjects, the salivary flow rates with splints were significantly higher than those without splints during relaxing, clenching, and chewing-like movement. The salivary flow rate during the chewing-like movement was significantly higher than that during relaxing and clenching, irrespective of splint application. The results suggest that maxillary occlusal splints might stimulate salivary secretion, particularly during chewing-like movement, in both bruxism patients and normal subjects. PMID- 15356503 TI - Nonsurgical correction of skeletal deep overbite and class II division 2 malocclusion in an adult patient. AB - Treatment modalities for Class II Division 2 malocclusion include growth modification, dental compensation, and surgical-orthodontic therapy; which treatment is chosen depends on the patient's age and growth potential. Deep overbite can be corrected by intrusion of anterior teeth, extrusion of posterior teeth, or a combination of both. Treatment considerations include the patient's facial profile, skeletal pattern, growth potential, and severity of dental malocclusion. Here, we present the nonsurgical orthodontic treatment of an adult patient with deep overbite and underlying skeletal Class II discrepancy. He had a hypodivergent facial pattern, Class II Division 2 malocclusion, and traumatic deep overbite due to supereruption of the mandibular anterior teeth. He refused orthognathic surgery but would accept orthodontic treatment alone, with the understanding that the treatment results could be compromised. We corrected the deep overbite by proclining the mandibular incisors; this helped to level the exaggerated curve of Spee. The posttreatment occlusion was significantly improved, both functionally and esthetically, with stable interincisal contacts. However, the improvement in occlusion and esthetics was achieved at the expense of reduced periodontal support for the mandibular anterior teeth. PMID- 15356504 TI - Use of digital photography in the Case orthodontic clinic. AB - In 2002, the orthodontic clinic at Case Western Reserve University totally converted to digital photography. We want to share the learning curve during this transition with clinicians planning the same change. A system and a protocol were developed for this transition; they have been in use for over a year. This system allows the handling of digital cameras when there are more clinicians than cameras; it can be applied to various specialties or fields. PMID- 15356505 TI - Litigation, legislation, and the ethics. The corporate practice of medicine. PMID- 15356506 TI - ABO invites question for phase II examination. PMID- 15356508 TI - When does the bag of waters break? PMID- 15356535 TI - Hereditary and acquired angioedema: problems and progress: proceedings of the third C1 esterase inhibitor deficiency workshop and beyond. AB - Hereditary angioedema (HAE), a rare but life-threatening condition, manifests as acute attacks of facial, laryngeal, genital, or peripheral swelling or abdominal pain secondary to intra-abdominal edema. Resulting from mutations affecting C1 esterase inhibitor (C1-INH), inhibitor of the first complement system component, attacks are not histamine-mediated and do not respond to antihistamines or corticosteroids. Low awareness and resemblance to other disorders often delay diagnosis; despite availability of C1-INH replacement in some countries, no approved, safe acute attack therapy exists in the United States. The biennial C1 Esterase Inhibitor Deficiency Workshops resulted from a European initiative for better knowledge and treatment of HAE and related diseases. This supplement contains work presented at the third workshop and expanded content toward a definitive picture of angioedema in the absence of allergy. Most notably, it includes cumulative genetic investigations; multinational laboratory diagnosis recommendations; current pathogenesis hypotheses; suggested prophylaxis and acute attack treatment, including home treatment; future treatment options; and analysis of patient subpopulations, including pediatric patients and patients whose angioedema worsened during pregnancy or hormone administration. Causes and management of acquired angioedema and a new type of angioedema with normal C1-INH are also discussed. Collaborative patient and physician efforts, crucial in rare diseases, are emphasized. This supplement seeks to raise awareness and aid diagnosis of HAE, optimize treatment for all patients, and provide a platform for further research in this rare, partially understood disorder. PMID- 15356542 TI - Chronic urticaria: pathogenesis and treatment. AB - Patients previously designated as having chronic idiopathic urticaria are now divided into 2 groups: 40% to 50% with chronic autoimmune urticaria, and the remainder with chronic idiopathic urticaria. Patients in both groups may have concomitant angioedema (approximately 40%). The autoimmune subgroup has an association with antithyroid antibodies and is caused by IgG antibody to the alpha subunit of the IgE receptor (35% to 40%), usually reactive with unoccupied IgE receptors, or IgG antibody to IgE (5% to 10%). Complement activation augments histamine secretion by release of C5a. The IgG subclasses that appear to be pathogenic are IgG(1), IgG(3), and, to a lesser degree, IgG(4), but not IgG(2). Histology of chronic urticaria (both subtypes) reveals a perivascular non necrotizing infiltrate of CD4(+) lymphocytes consisting of a mixture of T(H)1 and T(H)2 subtypes, plus monocytes, neutrophils, eosinophils, and basophils. These cells are recruited as a result of interactions with C5a, cell priming cytokines, chemokines, and adhesion molecules. Suggested therapy for patients with severe disease involves the use of high-dose hydroxyzine or diphenhydramine when nonsedating antihistamines are ineffective, supplemented by H-2 antagonists and leukotriene antagonists. The most severe patient may require protracted treatment with low-dose alternate-day steroid or cyclosporine. Cyclosporine can be steroid sparing when side effects are encountered or when use of steroids is relatively contraindicated. Careful monitoring of blood pressure, BUN, creatinine, and urinalysis is required. PMID- 15356544 TI - Turning off signal transducer and activator of transcription (STAT): the negative regulation of STAT signaling. AB - Signal transducer and activator of transcription (STAT) proteins are a group of transcription factors that transmit signals from the extracellular milieu of cells to the nucleus. They are crucial for the signaling of many cytokines that are mediators of allergic inflammation. Considerable information is known about the activation of STATs and their role in gene transcription; comparably much less is known about how STAT signaling is regulated. Because STATs are critical for the induction of many genes crucial for the allergic cascade and immune host defense, understanding the regulation of these molecules will provide novel insights into allergic and immunodeficiency disorders and will likely identify new targets for therapeutic interventions. This review will summarize the current understanding of the regulation of STAT signaling, emphasizing recent observations. PMID- 15356546 TI - Role of gender and hormone-related events on IgE, atopy, and eosinophils in the Epidemiological Study on the Genetics and Environment of Asthma, bronchial hyperresponsiveness and atopy. AB - BACKGROUND: The pattern of asthma over the lifespan is different in male and female patients, but etiologic differences according to gender are only partially understood. In women, information regarding factors explaining perimenstrual asthma and the role of hormone-related aspects on asthma-related phenotypes is scanty. OBJECTIVE: To assess the relationships of eosinophils, IgE, and atopy with (1) asthma according to gender and age of onset and (2) hormone-related events. METHODS: Using data from the Epidemiological study on the Genetics and Environment of Asthma, Bronchial Hyperresponsiveness and Atopy, adults and children with asthma recruited in chest clinics (n=313) and first-degree relatives of patients with asthma (n=214) were compared with nonasthmatic controls (n=334) and first-degree relatives without asthma (n=595). RESULTS: Among asthmatic women, eosinophilia was significantly associated with perimenstrual asthma independently from age, smoking, and asthma severity (eosinophils/mm(3) 330 vs 194; P=.01). In nonasthmatic women, IgE level was significantly decreased (by half) and atopy decreased with menopause, and IgE increased with oral contraceptive use, independently from age and smoking. Considering both genders, the increase of eosinophil counts with asthma was significantly greater in women with childhood-onset asthma than in women with adulthood-onset or in men in general. No interaction between gender and asthma was observed for eosinophils in children and for IgE level and atopy in children and adults. CONCLUSION: Results suggest a role of hormone-related events on asthma-related traits and support the hypothesis of the role of eosinophils in the persistence and severity of asthma. PMID- 15356547 TI - Characteristics of children with asthma who are enrolled in a Head Start program. AB - BACKGROUND: Asthma is an increasing public health concern that disproportionally affects children. In 1998, the Centers for Disease Control identified children aged 0 to 4 years as the "driving force" behind climbing rates of asthma-related emergency department visits and hospitalizations. Despite the significant asthma burden in preschool children, few studies characterize this population. OBJECTIVE: This study identifies and characterizes children at risk for asthma who are enrolled in a local Head Start program. METHODS: Caregivers of 368 children aged 3 to 5 years who were identified by using an asthma survey were recruited. Data were collected on demographics, health care use and access, medication use, symptoms, and trigger exposure. Exposure to tobacco smoke was determined by urinary cotinine and allergen sensitization by skin prick test. RESULTS: At baseline, 64% of the children had more than 1 emergency department visit for asthma in their lifetime, whereas 31% had more than 1 visit in the previous 6 months. Caretakers reported smoking in 37% of households with cotinine exceeding 20 ng/mg in 27% of the sample. Twenty-one percent reported symptoms consistent with intermittent asthma, and 79% reported symptoms consistent with persistent asthma. Forty-five percent of the children reported nighttime symptoms more than 1 night per week. Seventy-one percent had positive test results for more than 1 allergen, and 42% had positive test results for more than 3 allergens. Only 32% of children with persistent asthma had both rescue and controller medications. CONCLUSION: Children with asthma enrolled in a Head Start program have significant environmental tobacco smoke exposure, are highly atopic and symptomatic, and do not receive appropriate medication treatment. Overall, children in the study had poor asthma control. This high-risk group could benefit from case management programs. PMID- 15356548 TI - Improved bronchodilator effect of deep inhalation after allergen avoidance in asthmatic children. AB - BACKGROUND: In healthy adults and children, deep inhalation (DI) is able to reverse induced bronchoconstriction. This ability is impaired in asthma, but the reasons are still to be elucidated. OBJECTIVES: This study investigated whether the bronchodilator effect of DI during methacholine-induced bronchoconstriction can be improved by allergen avoidance in asthmatic children, and its relationship with airway inflammation. METHODS: The effect of DI on methacholine-induced bronchoconstriction was studied at the beginning and the end of a 3-month allergen avoidance period at high altitude in 14 allergic asthmatic children who had severe asthma attacks. Changes in airway caliber were inferred from the respiratory resistance (Rrs) measured by a forced oscillation technique. Results were related to the percentage of eosinophils in induced sputum and compared with those obtained in 9 age-matched nonasthmatic children. RESULTS: In asthmatic subjects, DI had no significant effect on methacholine-induced increase in Rrs before (P=.62) but significantly reversed it after (P <.01) allergen avoidance. However, the ability of DI to reverse a methacholine-induced increase in Rrs tended to remain less in asthmatic than nonasthmatic children even after allergen avoidance (P=.05). In the asthmatic children, the percentage of eosinophils in induced sputum was decreased at the end of the allergen avoidance period (P <.001), without any significant correlation between sputum eosinophils and airway responsiveness to methacholine or effect of DI. CONCLUSION: A short period of allergen avoidance may improve the ability of DI to reverse induced bronchoconstriction in some asthmatic children. This effect is associated, yet not correlated, with a reduction in airway inflammation. PMID- 15356549 TI - Exhaled nitric oxide as a marker of airway inflammation for an epidemiologic study in schoolchildren. AB - BACKGROUND: Exhaled nitric oxide (eNO) levels are increased in airway inflammatory disorders, such as asthma. OBJECTIVE: We sought to determine whether eNO could serve as a noninvasive marker of allergic airway inflammation for an epidemiologic study in schoolchildren. METHODS: Two hundred seventy-eight schoolchildren aged 10 to 12 years answered a modified American Thoracic Society questionnaire, and eNO levels and pulmonary function were measured. In 246 subjects serum nonspecific IgE levels and levels of IgE specific to house dust mite, cat, cedar, and mold were also measured. Correlation analysis was used to examine eNO levels, nonspecific IgE levels, antigen-specific IgE levels, and pulmonary function. In addition, we compared these variables between subjects with (recurrent wheezers) and without (nonwheezers) recurrent wheeze. Finally, multiple logistic regression analysis was used to find possible predictors for recurrent wheezers. RESULTS: eNO showed significant positive correlations with nonspecific IgE (r=0.62, P <.001) and mite-specific IgE (r=0.74, P <.001) and weak positive correlations with specific IgE to cat and cedar. Only eNO showed a weak but significant inverse correlation with pulmonary function (%FEV(1), P=.035; FEV(1)/forced vital capacity, P=.018). eNO, nonspecific IgE, and mite specific IgE levels in recurrent wheezers were greater (P <.001), and %FEV(1) was less (P=.06) when compared with values seen in nonwheezers. Finally, eNO was determined by means of multiple logistic regression analysis to be the best predictor for recurrent wheezers compared with other variables (odds ratio, 11.2; 95% CI, 1.33-94.0). CONCLUSION: eNO can be used in epidemiologic studies as a noninvasive marker of allergic airway inflammation in schoolchildren. PMID- 15356550 TI - Dosimeter methacholine challenge: comparison of maximal versus submaximal inhalations. AB - BACKGROUND: Deep inhalation has bronchodilating and bronchoprotective effects, particularly in subjects who are normal or have mild airway hyperresponsiveness (AHR). We have anecdotally observed that the 5 breath to total lung capacity (TLC) dosimeter method reduced the response to methacholine in some subjects with mild AHR. OBJECTIVE: To compare prospectively submaximal inhalations with TLC inhalations during the dosimeter methacholine challenge. METHODS: Sixteen subjects with asthma and a methacholine PC 20 <8 mg/mL performed 2 methacholine challenges in random order; the standard dosimeter method was compared with a modified dosimeter challenge in which methacholine inhalations were performed to approximately 50% to 60% below TLC. RESULTS: The standard methacholine challenge PC 20 was almost twice that obtained with the modified submaximal inhalation method (geometric mean PC 20, 5.2 mg/mL vs 2.8 mg/mL, respectively; P = 0.0216). In the 5 subjects with the mildest AHR, there was a 2.5-fold to 14-fold difference in PC 20 between methods. The standard (full TLC) PC 20 s were falsely negative (>16 mg/mL) in these 5 subjects with current asthma, 4 of whom required inhaled corticosteroids. CONCLUSION: A submaximal inhalation dosimeter methacholine challenge results in a significantly lower PC 20 compared with the standard 5-breath dosimeter method. This effect is limited to the mildly responsive group, probably because of the bronchoprotective effect of the deep inhalation during the standard method, and results in false-negative tests in some subjects. PMID- 15356551 TI - Natriuretic peptides and genesis of asthma: an emerging paradigm? AB - Exposure to allergens and infections contribute to early immune development. However, knowledge of the role of cellular metabolic, physiologic, and endocrinologic factors in controlling immune development and asthma is limited. Immune cells, including macrophages, dendritic cells, and T lymphocytes, express receptors for atrial natriuretic peptide (ANP) both in the fetal and neonatal lymphoid organs. ANP has garnered much attention for its cardiovascular effects, but its apparently significant role in the physiology and immunity of the lung has been underappreciated. Studies indicate that ANP also plays a significant role in shaping the early immune responses to environmental antigens. The C terminal prohormone natriuretic peptide ANP (or NP(99-126)), which possesses bronchodilatory properties, is involved in polarizing dendritic cells to produce a T(H)2 response. Also, de novo overexpression of another pro-ANP peptide, NP(73 102), provides persistent bronchoprotection and induces significant anti inflammatory activities in the lung epithelial cells. Thus natriuretic peptides appear to play a pivotal role in the genesis and control of asthma, and they might provide an important target to modulate allergen-induced immune responses in allergic patients. PMID- 15356553 TI - Umbilical cord and maternal blood red cell fatty acids and early childhood wheezing and eczema. AB - BACKGROUND: Few studies have explored whether fetal exposure to n-6 and n-3 fatty acids influences the inception of atopic disease. OBJECTIVE: To assess prenatal fatty acid exposures as predictors of early childhood wheezing and eczema. METHODS: In the Avon Longitudinal Study of Parents and Children, late pregnancy maternal blood samples and umbilical cord blood samples were assayed for n-6 and n-3 fatty acids (percentage of total red cell phospholipid), and mothers were asked about wheezing and eczema in their children. We measured associations of 11 n-6 and n-3 fatty acid exposures with wheezing at 30 to 42 months, with wheezing patterns defined by presence (+) or absence (-) of wheezing during 2 periods, 0 to 6 months and 30 to 42 months (transient infant, +/-; later-onset, -/+; persistent, +/+; n=1191 and n=2764 for cord and maternal analyses, respectively), and with eczema at 18 to 30 months (n=1238 and n=2945 for cord and maternal analyses, respectively). RESULTS: In cord blood red cells, the ratio of arachidonic:eicosapentaenoic acid was positively associated with eczema (adjusted odds ratio [OR] per doubling, 1.14; 95% CI, 1.00-1.31; P=.044), the ratio of linoleic acid:alpha-linolenic acid was positively associated with later-onset wheeze (OR, 1.30; CI, 1.04-1.61; P=.019), and the ratio of alpha-linolenic acid:n 3 products was negatively associated with later-onset wheeze (OR, 0.86; CI, 0.75 0.99; P=.040). However, these associations were no longer significant after adjusting for multiple comparisons. CONCLUSIONS: It seems unlikely that fetal exposure to n-6 and n-3 fatty acids is an important determinant of early childhood wheezing and atopic disease. PMID- 15356552 TI - Omalizumab-induced reductions in mast cell Fce psilon RI expression and function. AB - BACKGROUND: By design, omalizumab binds free IgE in the circulation and prevents its attachment to the surface of mast cells and basophils, thereby preventing them from responding to allergens. Previously, omalizumab rapidly reduced free IgE levels, as well as basophil high-affinity IgE receptors, leading to significant reductions in basophil mediator response to allergen. It is assumed that tissue mast cells are similarly altered in their Fc epsilon RI density and function. OBJECTIVE: We examined the phenotypic shift of skin mast cells in parallel to that of blood basophils in 3 subjects infused with omalizumab. METHODS: Three subjects with allergic rhinitis underwent intradermal skin test titration with house dust mite antigen at days 0, 7, 70, and 196 of omalizumab treatment. As control subjects, 5 untreated subjects with allergic rhinitis were evaluated at similar time points. All subjects underwent skin biopsy 18 to 24 hours later at the site of allergen injection. Biopsy specimens were characterized by means of immunohistochemisty for tryptase and Fc epsilon RI alpha immunoreactivity, as well other markers (CD3, CD45RO, CD68, cutaneous lymphocyte antigen, and major basic protein). RESULTS: Omalizumab recipients, but not control subjects, demonstrated reductions in Fc epsilon RI alpha immunoreactivity at days 70 and 196 in parallel with reductions in the acute wheal response to allergen. However, no reductions in tryptase-positive cells were noted at these time points. CONCLUSION: Reductions in free IgE levels by omalizumab leads to a rapid reduction in basophil Fc epsilon RI receptor expression. In contrast, the time course for the decrease of Fc epsilon RI expression in skin mast cells is slower and associated with decreased acute allergen wheal size. PMID- 15356554 TI - Effects of antiasthmatic agents on the functions of peripheral blood monocyte derived dendritic cells from atopic patients. AB - BACKGROUND: Dendritic cells (DCs), the antigen-presenting cells in the airway, play a critical role in asthma. Nevertheless, there is little information on the effects of antiasthmatic agents on DCs. OBJECTIVES: The purpose of the present study was to determine the effects of representative antiasthmatic agents, including cysteinyl leukotriene (cysLT) 1 receptor antagonists, corticosteroid, and tacrolimus, on DCs in inducing allergy. METHODS: Human peripheral blood monocyte-derived DCs (MoDCs) generated from atopic and healthy subjects were pulsed with Dermatophagoides farinae allergen in the presence of medium alone, pranlukast, montelukast, dexamethasone, or tacrolimus. The mRNA expressions of cysLT receptor, cysLTs producing enzymes, and various surface markers on MoDCs, as well as the concentrations of cysLTs, IL-10, and IL-12 in cultured supernatants, were determined. MoDCs were also cocultured in vitro with autologous CD4(+) T cells, and IL-5 and IFN-gamma production was measured. RESULTS: MoDCs of atopic patients expressed mRNAs of cysLT1 receptor and cysLT producing enzymes, and allergen pulsing significantly increased cysLT production. MoDCs of atopic patients showed a T(H)2-favoring phenotype and induced T(H)2 skewed cytokine production from autologous CD4(+) T cells. Dexamethasone and tacrolimus inhibited allergen-pulsed MoDC-induced cytokine production by autologous CD4(+) T cells without and with IL-10 inhibition, respectively. CysLT1 receptor antagonists had no effect on MoDC functions. CONCLUSION: Our results indicate that MoDCs of atopic patients induce a T(H)2 reaction. Corticosteroid and tacrolimus, but not cysLT1 receptor antagonists, inhibit T(H)2 reactions, and this effect is probably mediated through different pathways. PMID- 15356555 TI - Heat denaturation affects the ProDer p 1 IgE reactivity and downregulates the development of the specific allergic response. AB - BACKGROUND: Modified allergens with reduced IgE-binding activity represent an elegant approach to circumvent the risk of anaphylactic reactions in allergen specific immunotherapy. OBJECTIVE: The current work investigated the effect of heat denaturation on the allergenic properties of recombinant ProDer p 1, a precursor form of the major house dust mite allergen Der p 1. METHODS: The IgE reactivity was estimated by direct and competition ELISA. The immunogenicity of heat-denatured ProDer p 1 was evaluated in naive and Der p 1-allergic mice. RESULTS: Heat denaturation in reducing conditions drastically reduced the in vitro ProDer p 1 IgE reactivity toward human allergic sera. In naive mice, heat denatured ProDer p 1 generated mixed T(H)1-T(H)2 responses characterized by the absence of specific IgE with concomitant rise in specific IgG2a titers and the presence of IL-5 and IFN-gamma in splenocyte cultures. In contrast, natural Der p 1 or native ProDer p 1 induced typical strict T(H)2-biased allergic responses with strong IgG1 and IgE titers, whereas spleen cells exhibited only high IL-5 secretion. Moreover, native or heat-denatured ProDer p 1 vaccinations prevented airway eosinophil infiltrations after challenge. Although native or heat-treated ProDer p 1 adjuvanted with SBAS1b induced mixed T(H)1-T(H)2 responses in allergic mice, heat-denatured ProDer p 1, compared with native ProDer p 1, proved to be more effective in redirecting the T(H)2-allergic response toward T(H)1. CONCLUSION: Taken together, our results suggest that variants of Der p 1 with reduced IgE-binding reactivity could represent hypoallergenic molecules suitable for allergen-specific immunotherapy. PMID- 15356556 TI - Functional effect of the R110Q IL13 genetic variant alone and in combination with IL4RA genetic variants. AB - BACKGROUND: IL-13 is a key mediator of allergic asthma. IL-13 mediates its effects via its receptor, a heterodimer composed of IL-4R alpha and IL-13R alpha1. Polymorphic variants of both IL-13 and IL-4R alpha have been shown to be associated with atopy. OBJECTIVE: We examined the functional consequences of the Q110 IL-13 variant in vitro and in vivo to determine whether it displays enhanced functional activity compared with R110 IL-13, both in the context of I50Q551 IL 4R alpha and of the atopy-associated variant V50R551 IL-4R alpha. METHODS: We used a mouse cell line stably expressing human IL-4R alpha and IL-13R alpha1 that readily responds to human IL-4 and IL-13. For in vivo analyses, we used BALB/c mice. RESULTS: The Q110 IL-13 variant displayed significantly increased activity compared with R110 IL-13. Furthermore, mice treated with Q110 IL-13 variant displayed increased airways hyperresponsiveness relative to R110 IL-13. We then examined the functional consequences of Q110 IL-13 variant in combination with an atopy-associated variant of its receptor, IL-4R alpha (V50R551). Q110 IL-13 variant had increased activity on these cells as well, and, strikingly, the effect was greater than that observed in cells expressing I50Q551 IL-4R alpha. CONCLUSION: Either Q110 IL-13 variant or V50R551 IL-4R alpha variant has enhanced function alone, but the 2 together have a synergistic effect on IL-13-dependent gene induction. Our data demonstrate the importance of relatively small individual differences in gene products from common single nucleotide polymorphisms that may result in larger combined differences. Furthermore, a relatively modest change in function from a single nucleotide polymorphism can result in an important biological difference in vivo. PMID- 15356557 TI - A TLR4 polymorphism is associated with asthma and reduced lipopolysaccharide induced interleukin-12(p70) responses in Swedish children. AB - BACKGROUND: Bacterial signals play an important role in the maturation of the immune system. Polymorphisms in genes coding for receptors to bacterial components can alter the immune responsiveness of the host to microbial agents and may indicate the development of aberrant immune responses that are associated with immune-mediated diseases such as atopic diseases. OBJECTIVE: The study's objective was to investigate the relationship between TLR4 and CD14 gene polymorphisms, the LPS responsiveness of PBMCs, and the presence of asthma and allergic rhinoconjunctivitis in children. METHODS: The TLR4 (Asp299Gly) and CD14/ 159 polymorphisms were determined in 115 Swedish children aged 8 and 14 years. LPS-induced IL-12(p70), IL-10, and IFN-gamma responses of PBMCs from 69 of the children were analyzed by means of ELISA. The levels of soluble CD14 in serum samples were analyzed by means of ELISA, and the total IgE levels were analyzed by means of UniCAP Total IgE (Pharmacia Diagnostics, Uppsala, Sweden). RESULTS: Decreased LPS-induced IL-12(p70) and IL-10 responses were associated with the TLR4 (Asp299Gly) polymorphism and independently with asthma, especially atopic asthma. The TLR4 (Asp299Gly) polymorphism was associated with a 4-fold higher prevalence of asthma in school-aged children (adjusted odds ratio 4.5, 95% CI 1.1 17.4) but not to allergic rhinoconjunctivitis. CONCLUSION: A TLR4 polymorphism modifies innate immune responses in children and may be an important determinant for the development of asthma. This may influence the outcome of intervention studies that use microbial stimuli as immune modulators. PMID- 15356558 TI - Peripheral blood eosinophils from patients with allergic asthma contain increased intracellular eosinophil-derived neurotoxin. AB - BACKGROUND: One mechanism of the eosinophil's contribution to airway inflammation in asthma is through release of cationic granule proteins to cause airway injury. Differences in either the intracellular concentration of granule proteins or the extent of activated degranulation between eosinophils from healthy patients and those with allergy and asthma could, therefore, relate to fundamental differences in this cell's function. OBJECTIVE: To identify phenotypic differences in eosinophil-derived neurotoxin (EDN) content and release in eosinophils from healthy patients, those with allergy, and those with allergy and asthma. METHODS: Peripheral blood eosinophils were isolated by negative anti-CD16 selection. Total intracellular and cytokine-activated release of EDN protein was measured by radioimmunoassay. EDN mRNA was assessed by real-time PCR. RESULTS: Eosinophils from patients with asthma contained significantly more EDN per cell than comparable cells from healthy patients, those with allergy but without asthma, or those with asthma treated with inhaled corticosteroids, but they had concentrations similar to airway eosinophils isolated from bronchoalveolar lavage fluid 48 hours after segmental bronchoprovocation with allergen. Furthermore, this increased granule protein was reflected in more EDN degranulation by IL-5- or GM-CSF-activated eosinophils when calculated as nanograms of protein secreted but not when calculated as a percentage of total EDN release. Levels of EDN mRNA were similar in all subject groups. CONCLUSIONS: These data suggest that peripheral blood eosinophils from subjects with untreated asthma have increased inflammatory capacity, as reflected by greater intracellular concentrations of EDN. PMID- 15356559 TI - Safety and application of induced sputum analysis in childhood asthma. AB - BACKGROUND: The value of sputum induction in pediatric asthma lies in its potential to directly and noninvasively assess airway inflammation in children, because bronchoscopy and biopsy carry some risk. The Childhood Asthma Management Program (CAMP) study was designed to evaluate the long-term effects of budesonide and nedocromil compared with placebo in children with mild to moderate asthma across 8 centers. OBJECTIVE: At the Denver CAMP site, we sought to evaluate the safety of sputum induction, to determine differences in airway inflammation between treatment groups by using induced sputum analysis, and to examine correlations between other biomarkers and sputum eosinophils. METHODS: Sputum induction was performed, and exhaled nitric oxide, circulating eosinophil counts, and serum eosinophil cationic protein were obtained at treatment discontinuation and after washout. Spirometry and a methacholine challenge were also performed according to the CAMP protocol. RESULTS: Ninety of 117 children provided an adequate sputum sample for analysis. In 9 subjects (3 nedocromil and 6 placebo), sputum induction resulted in bronchospasm. These subjects had greater disease severity, as measured by a lower median prebronchodilator FEV 1 percentage predicted (85.0% vs 96.0%; P =.024) and FEV 1 /FVC ratio (70.0% vs 79.0%; P =.0008); greater bronchodilator reversibility (16.5% vs 6.8%; P =.004); higher serum IgE (1390.0 vs 495.0 ng/mL; P =.017) and circulating eosinophil count (757.0 vs 282.0/mm 3; P =.04); greater use of prednisone (1.9 vs 0.9 courses per 100 person-years; P =.05); and greater supplemental inhaled steroid doses (85.3 vs 0 mg; P =.016). At treatment discontinuation, budesonide-treated patients had a lower median (1st, 3rd quartile) sputum percentage eosinophil (SPEos) (0.2% [0%, 1.2%] vs 0.8% [0.2%, 4.6%]; P =.03) compared with those treated with placebo; no significant difference was noted between nedocromil- and placebo treated patients. Higher SPEos at the time of treatment discontinuation was associated with asthma worsening that required rescue prednisone (n = 23) during the washout period compared with patients who remained stable (3.6% [0.4%, 6.4%] vs 0.6% [0.2%, 3.2%] SPEos; P =.023). Finally, greater SPEos was associated with atopy, higher bronchodilator reversibility, lower FEV 1 /FVC ratio, higher exhaled nitric oxide levels, circulating eosinophils, sputum and serum eosinophil cationic protein, more prednisone courses during the treatment period, and greater asthma severity. CONCLUSIONS: Sputum induction is a relatively noninvasive and safe procedure that can provide information on eosinophilic inflammation and treatment response and is also associated with several measures of asthma control. However, this procedure still remains a research tool in asthma because of its requirements for technical expertise. PMID- 15356560 TI - LPS-binding protein as a target molecule in allergy and asthma. PMID- 15356561 TI - LPS binding protein is important in the airway response to inhaled endotoxin. AB - BACKGROUND: Inhaled endotoxin is a risk factor for asthma exacerbation, and endotoxin inhalation by itself recapitulates many of the classical features of asthma in mice, including reversible airflow obstruction and inflammation, airways hyperresponsiveness, and airway remodeling. OBJECTIVE: Our objective was to determine the importance of LPS binding protein (LBP) in the response to inhaled LPS. METHODS: We challenged LBP-deficient mice (C57BL/6(LBP-/-)) and C57BL/6 mice with inhaled endotoxin for 4 hours, 5 days, or 4 weeks, followed by 3 days of recovery. RESULTS: LBP in the lung was significantly increased in LPS exposed C57BL/6 mice from all 3 groups. Only LPS-exposed C57BL/6 mice had significantly enhanced airway responsiveness to inhaled methacholine. Total lavage cells in LPS-exposed C57BL/6(LBP-/-) mice were significantly reduced compared with those seen in LPS-exposed C57BL/6 mice; however, the percentage of PMNs was similarly increased in both the C57BL/6 and C57BL/6(LBP-/-) mice. TNF alpha, IL-1 beta, and IL-6 protein concentrations in whole-lung lavage fluid from C57BL/6(LBP-/-) mice were also significantly reduced when compared with those seen in C57BL/6 mice. In C57BL/6(LBP-/-) mice submucosal cell proliferation was significantly reduced in the 1-week group when compared with that seen in similarly exposed C57BL/6 mice. The 4-week exposed C57BL/6 mice had significantly thickened airway submucosa and significantly increased lavaged TGF-beta(1) protein compared with that seen in C57BL/6(LBP-/-) mice. CONCLUSIONS: These findings indicate that LBP is one of the critical molecules regulating the acute and chronic airway response to inhaled LPS. PMID- 15356563 TI - Hev b 5 and Hev b 13 as allergen markers to estimate the allergenic potency of latex gloves. AB - BACKGROUND: Sensitization to natural rubber latex has been linked to proteins from medical latex gloves. Various assays to estimate the amount of residual allergenic proteins extractable from latex gloves to assess their potential exposure hazard have inherent weaknesses. OBJECTIVE: This investigation was aimed at developing 2-site immunoenzymetric assays and identifying appropriate protein markers to assess the allergenic potential of latex gloves. METHODS: The presence of 6 latex allergens--Hev b 1, 2, 3, 5, 6, and 13--was measured in a cross section of commercial latex medical gloves by using monoclonal and polyclonal antibody-based 2-site immunoenzymetric assays. The overall allergenic potential of these gloves was assessed by IgE-inhibition assay. Stepwise multiple regression analyses were performed to identify marker allergens that best explained the variation in latex glove allergenicity. RESULTS: All 6 latex allergens were detected in at least some of the glove samples. Hev b 5 and Hev b 13 were identified as the marker allergens that combined best to explain the variation in the glove allergenicity. The significant multiple correlation (R=0.855) between these 2 markers and glove allergenic potency forms the basis of an assay to gauge latex glove allergenicity. CONCLUSION: The overall allergenic potential of latex gloves can be estimated by using Hev b 5 and Hev b 13 as indicator allergens. The correlation between glove allergenicity and the level of these allergens was maintained for low-protein gloves (<200 microg/g). This estimation of glove allergenicity was superior to that obtained by using total protein readings. PMID- 15356564 TI - Airborne fungi in the homes of children with asthma in low-income urban communities: The Inner-City Asthma Study. AB - BACKGROUND: Despite growing evidence of the importance of exposure to fungi as an environmental risk factor for asthma, few data have been reported on the exposure to airborne fungi of asthmatic children living in US inner cities. OBJECTIVE: We sought to examine the spectrum and concentration of fungi in the air inside and outside of the homes of mold-sensitive children with asthma living in US cities. We also analyzed the relationship of the concentration of fungi in indoor air to home characteristics. METHODS: We performed a home environmental survey and measured the concentrations of culturable airborne fungi inside and outside the homes of 414 mold-sensitive children with asthma in 7 urban communities. RESULTS: The airborne fungi encountered indoors generally paralleled those found outdoors, and the similarities between communities were more striking than the differences. Indoor fungal concentrations were correlated with outdoor concentrations measured on the same day, suggesting the need to adjust for the outdoor concentration in analyses by using the indoor concentration as an indicator of the relative moldiness of a home. The concentration of fungi in indoor air in excess of outdoor air-that is, the indoor-outdoor difference-was significantly related to home characteristics, including dampness, having a cat, and cockroach infestation. CONCLUSION: Mold-sensitive children with asthma living in urban communities across the US are exposed to airborne fungi in indoor and outdoor air. The concentrations of fungi are higher in homes with dampness problems, cockroach infestation, and cats. The indoor-outdoor difference in the concentration of airborne fungi may provide a valuable metric for investigations of the role of fungal exposure as a risk factor for asthma. PMID- 15356565 TI - Preventing peptide-induced anaphylaxis: addition of C-terminal amino acids to produce a neutral isoelectric point. AB - BACKGROUND: Progress in peptide immunotherapy for the treatment of autoimmune diseases has been hampered by reports of anaphylactic reactions in both mice and human subjects. Fatal anaphylaxis in nonobese diabetic (NOD) mice has been described after repeated subcutaneous insulin peptide B:9-23 immunizations. On the basis of observations that rapid systemic delivery of peptide to a sensitized mouse (eg, intravenous delivery) increases the anaphylactic response, it was hypothesized that slowing down the absorption of the peptide would prevent anaphylaxis. OBJECTIVES: We sought to prevent anaphylaxis from B:9-23 peptide by altering the isoelectric point (pI) to neutral, thereby decreasing solubility and rate of absorption after subcutaneous injection. METHODS: B:9-23 peptide was modified by the addition of 2 arginine (RR) amino acids to the C-terminus to create B:9-23RR, thereby increasing the pI from 5.4 to 7.0. Both native and modified B:9-23 peptide were tested for the ability to induce anaphylaxis in a NOD mouse model of self-peptide anaphylaxis. RESULTS: This modification resulted in a peptide vaccine with decreased solubility when administered subcutaneously at a neutral pH. B:9-23RR significantly protected NOD mice from peptide-induced anaphylaxis compared with B:9-23 peptide. Furthermore, B:9-23RR peptide retains its ability to induce insulin autoantibodies and prevent diabetes in NOD mice. CONCLUSION: The modification of the pI of a peptide vaccine might be a generalizable method to prevent anaphylaxis without changing the immunologic properties. PMID- 15356566 TI - Peanut protein allergens: gastric digestion is carried out exclusively by pepsin. AB - BACKGROUND: A major characteristic of many food allergens, including Ara h 1, a major peanut allergen, is their resistance to gastric digestion. One estimate of the allergenic potential of a possible protein allergen is its stability under simulated gastric conditions. OBJECTIVE: Because the rate and extent of digestion of allergenic proteins will affect the severity of any subsequent allergic response, it is important to correlate protein allergen digestion in simulated gastric fluid with that in actual gastric fluid. METHODS: A major peanut allergen, Ara h 1, was digested in vitro by using both pepsin and porcine gastric fluid. Several comparisons between the 2 sets of proteolytic conditions were assessed including pH optima and the effect of temperature, denaturants, and specific enzyme inhibitors. RESULTS: In vitro digestion of Ara h 1 with pepsin and porcine gastric fluid resulted in virtually identical hydrolysis patterns as observed on SDS-PAGE. The protease activity of both pepsin and gastric fluid were inhibited at high pH and in the presence of pepstatin. However, both remained active in 4 mol/L urea and at 60 degrees C. CONCLUSIONS: Protein digestion in the porcine stomach is carried out by pepsin. In vivo gastric digestion is modeled accurately by peptic hydrolysis. Digestion conditions in vivo are comparable to experimental conditions in vitro provided that the acidic nature of the stomach contents is optimal for characterization of the allergen under standard pepsin digestion conditions. Additional experimentation using crude food extracts, both in the presence and absence of a complete meal, is needed to elucidate the complete physiologic nature of food allergen digestion. PMID- 15356567 TI - Randomized placebo-controlled trial comparing desloratadine and montelukast in monotherapy and desloratadine plus montelukast in combined therapy for chronic idiopathic urticaria. AB - BACKGROUND: H 1 -receptor antagonists are considered to be particularly effective in reducing pruritus, and they are therefore recommended as first-line treatment in patients with chronic idiopathic urticaria (CIU). Recently, antileukotriene receptors have been used in patients with CIU, either administered as monotherapy or combined with H 1 -receptor antagonists. OBJECTIVE: We compared the clinical efficacy of 5 mg of desloratadine administered once daily either as monotherapy or combined with a leukotriene antagonist, 10 mg of montelukast daily, and 10 mg of montelukast administered daily as monotherapy for the treatment of patients affected by CIU with placebo. METHODS: One hundred sixty patients aged 18 to 69 years (mean +/- SD, 43.9 +/- 13.4 years) with a history of moderate CIU were selected. A randomized, double-blind, double-dummy, placebo-controlled, parallel group study design was used. Patients were treated with 5 mg of desloratadine once daily (n = 40), 10 mg of montelukast once daily (n = 40), 5 mg of desloratadine (n = 40) in the morning plus montelukast in the evening, or matched placebo (n = 40). Assessment of treatment efficacy was based on scores of daily cutaneous symptoms evaluated reflectively and instantaneously. RESULTS: Only the group treated with desloratadine as monotherapy or as combined therapy concluded the whole study. Twenty-seven of the 40 patients in the montelukast group and 35 of the 40 patients in the placebo group discontinued the treatment. As reflective evaluation, all groups showed significant differences compared with the placebo group in terms of total symptom score, number of hives, and size of largest hive. In addition to the pruritus, only the groups treated with desloratadine as monotherapy or combined therapy showed significant differences compared with those receiving placebo, whereas there were no differences between the montelukast and placebo groups. Finally, no differences were found between the desloratadine group and the desloratadine plus montelukast group. The instantaneous evaluation demonstrated similar results regarding the desloratadine group and the desloratadine plus montelukast group versus the placebo group, whereas there were no significant differences between the group treated with montelukast alone and the placebo group for pruritus and size of largest hive. No differences were found between the group treated with desloratadine alone and the desloratadine plus montelukast group. CONCLUSIONS: The results of this comparative study demonstrate that desloratadine is highly effective for the treatment of patients affected by CIU. In addition, the regular combined therapy of desloratadine plus montelukast does not seem to offer a substantial advantage with respect to desloratadine as monotherapy in patients affected by moderate CIU. PMID- 15356568 TI - Hereditary angioedema: a half century of progress. PMID- 15356569 TI - Canadian 2003 International Consensus Algorithm For the Diagnosis, Therapy, and Management of Hereditary Angioedema. AB - C1 inhibitor deficiency (hereditary angioedema [HAE]) is a rare disorder for which there is a lack of consensus concerning diagnosis, therapy, and management, particularly in Canada. European initiatives have driven the approach to managing HAE with 3 C1-INH Deficiency Workshops held every 2 years in Hungary starting in 1999, with the third Workshop having recently been held in May 2003. The European Contact Board has established a European HAE Registry that will hopefully advance our knowledge of this disorder. The Canadian Hereditary Angioedema Society/Societe d'Angioedeme Hereditaire du Canada organized a Canadian International Consensus Conference held in Toronto, Ontario, Canada, on October 24 to 26, 2003, to foster consensus between major European and North American HAE treatment centers. Papers were presented by investigators from Europe and North America, and this consensus algorithm approach was discussed. There is a paucity of double-blind placebo-controlled trials in the treatment of HAE, making levels of evidence to support the algorithm less than optimal. Enclosed is the consensus algorithm approach recommended for the diagnosis, therapy, and management of HAE and agreed to by the authors of this article. This document is only a consensus algorithm approach and requires validation. As such, participants agreed to make this a living 2003 algorithm (ie, a work in progress) and agreed to review its content at future international HAE meetings. The consensus, however, has strength in that it was arrived at by the meeting of patient-care providers along with patient group representatives and individual patients reviewing information available to date and reaching agreement on how to approach the diagnosis, therapy, and management of HAE circa 2003. Hopefully evidence to support approaches to the management of HAE will approach the level of meta-analysis of randomized controlled trials in the near future. PMID- 15356570 TI - C1 inhibitor gene expression in patients with hereditary angioedema: quantitative evaluation by means of real-time RT-PCR. AB - BACKGROUND: Hereditary angioedema (HAE) is caused by heterozygous defects in the C1 inhibitor (C1-INH) gene (SERPING1/C1NH). In patients' plasma C1-INH levels range between 5% and 30% of normal levels (ie, far from the 50% expected for an autosomal dominant defect). Most patients have antigenic and functional deficiency (type I HAE), and 15% have reduced C1-INH function but normal to increased antigen because of the presence of a dysfunctional protein (type II HAE). OBJECTIVE: We sought to contribute to the understanding of the pattern of C1-INH gene expression in patients with HAE. METHODS: We used real-time quantitative RT-PCR to measure C1-INH mRNA levels in PBMCs of 57 patients with HAE typed for mutations in the SERPING1/C1NH gene. RESULTS: Thirty-six different mutations were identified in genomic DNA. Compared with healthy control subjects, C1-INH mRNA was significantly and similarly reduced in patients with type I and type II HAE (40% and 47%, respectively; P <.0001). By means of direct sequencing of cDNAs, we found that 74% of patients with type I HAE carrying small mutations presented significant amounts of mutated transcripts at the mRNA level, suggesting that both allelic mRNA products were reduced to approximately 50%. In 4 patients carrying large deletions expected to fully inactivate expression from the mutant allele, C1-INH mRNA was 23% on average compared with that seen in control subjects, confirming that normal mRNA was strongly underexpressed. CONCLUSIONS: These new findings, combined with previous evidence of increased C1 INH consumption, might explain the plasma levels of normal C1-INH that are markedly less than the expected 50%. PMID- 15356571 TI - IL-4-independent pathways exacerbate methacholine-induced airway hyperreactivity during mycoplasma respiratory disease. AB - BACKGROUND: Mycoplasma respiratory infections are associated with wheezing and exacerbation of airway hyperresponsiveness (AHR) in asthmatic patients. IL-4 is a key cytokine in the development of AHR and airway reconstruction in asthmatic patients and might be an important component in exacerbation of AHR during pulmonary mycoplasma infection. OBJECTIVE: This study evaluates the effect of IL 4 on the severity of methacholine-induced AHR associated with mycoplasma pulmonary mycoplasma infection. METHODS: BALB/c and IL-4 knockout (KO) mice were infected with Mycoplasma pulmonis, and their enhanced pause scores were monitored before and after methacholine inhalation with whole-body plethysmography. RESULTS: IL-4 KO mice showed no difference in histopathology of the lungs before or after Mycoplasma pulmonis infection when compared with BALB/c mice. There was an increase in airway obstruction from days 7 to 21 after infection in both strains of mice, but there was no strain difference in airway resistance associated mycoplasma disease. However, IL-4 KO mice had significantly higher methacholine-induced AHR after M pulmonis infection when compared with BALB/c mice. There was no difference in AHR between uninfected IL-4 KO and control mice. CONCLUSION: In contrast to our hypothesis, IL-4-independent pathways exacerbate methacholine-induced AHR and promote airway obstruction during the pathogenesis of mycoplasma respiratory disease. PMID- 15356572 TI - Human nuclear factor kappa B essential modulator mutation can result in immunodeficiency without ectodermal dysplasia. AB - BACKGROUND: Many receptors rely on the appropriate activation of nuclear factor (NF) kappa B to induce cellular function. This process depends critically on the phosphorylation of the inhibitor of NF-kappa B (I kappa B) by the I kappa B kinase. This targets I kappa B for ubiquitination and degradation, allowing NF kappa B to translocate to the nucleus, where it can direct transcription. Hypomorphic human mutations affecting one I kappa B kinase component, the NF kappa B essential modulator (NEMO), result in impaired signaling from receptors required for ectodermal development and immune function. Male subjects with these mutant NEMO molecules have an X-linked syndrome known as ectodermal dysplasia with immunodeficiency, which is characterized by severe infections, with herpesviruses, bacteria, and mycobacterial susceptibility. OBJECTIVE: We sought to genetically and biochemically characterize a patient with a mutant NEMO molecule without ectodermal abnormalities. METHODS: We evaluated NEMO in a patient who had immunodeficiency and atypical mycobacterial infection but normal ectoderm. RESULTS: We identified a novel NEMO mutant causing immunodeficiency without ectodermal dysplasia. The mutation, which altered the exon 9 splice site, was present in cells of ectodermal and hematopoetic origin and resulted in a heterogeneous mixture of mutant and wild-type cDNA species. Immunologic function was variably impaired, with reduced CD40-induced B-cell proliferation, partially reduced NF-kappa B p65 nuclear translocation, and variable Toll-like receptor induced TNF production. This variability might be explained by an inconsistent ratio of mutant to wild-type NEMO. The lack of any ectodermal phenotype, however, suggested a separation in the hematopoetic and ectodermal function of NEMO that leads to NF-kappa B activation. CONCLUSION: Mutation of the gene encoding NEMO can result in immunodeficiency without ectodermal dysplasia. PMID- 15356573 TI - Synthesis of IL-13 by human B lymphocytes: regulation and role in IgE production. AB - BACKGROUND: Our laboratory has demonstrated previously that human tonsillar B lymphocytes express IL-13 mRNA OBJECTIVE: We sought to investigate IL-13 production by human B cells and the association between B cell-derived IL-13 and IgE secretion. METHODS: Human B lymphocytes were isolated from tonsils and purified by means of rosetting with sheep RBCs or positive or negative selection with magnetic beads. They were stimulated with anti-CD40 antibodies with or without recombinant IL-4. Total mRNA was extracted, and IL-13 mRNA was measured by means of standard RT-PCR or by means of real-time PCR with commercially available primers. B cells were cultured with or without IL-13 neutralizing antibodies, and C epsilon transcripts and supernatant IgE levels were measured. RESULTS: IL-13 mRNA was detected in human B lymphocytes stimulated with anti-CD40 antibodies and IL-4 or IL-2 but not in unstimulated B cells. Real-time PCR demonstrated a 10- to 15-fold increase in IL-13 mRNA, maximizing at 36 hours. IL 13 protein was detected from B lymphocytes on day 3 and accumulated through day 7. The synthesis of IL-13 required both CD40 and IL-4 stimulation. The presence of IL-13 was confirmed by means of intracellular staining of cultured B lymphocytes and antigen-stimulated nasal biopsy specimens from atopic individuals. Addition of IL-13 neutralizing antibodies to purified B-cell cultures inhibited IgE production by up to 80% and diminished IgE (C epsilon) transcripts by 50%. CONCLUSION: Human B lymphocytes express IL-13 mRNA after ligation of CD40 and the addition of cytokines. Human B lymphocytes produce significant IL-13, and neutralization of IL-13 impairs IgE synthesis. IL-13 might be an important autocrine growth factor for IgE-producing B lymphocytes. PMID- 15356574 TI - Regulation of TH2 responses by the pulmonary Clara cell secretory 10-kd protein. AB - BACKGROUND: Pulmonary Clara cell secretory 10-kd protein (CC10) is a steroid inducible and potentially anti-inflammatory cytokine, but its direct involvement in the regulation of T-cell responses remains unknown. OBJECTIVE: The role of CC10 in the regulation of T(H)2 cytokine expression was investigated. METHODS: The levels of cytokine and GATA-3 expression were determined by ELISA and RT-PCR, respectively. Bronchoalveolar lavage fluid cell counts were also determined by using a standard protocol. CC10 expression in vivo was determined by immunocytochemistry and Western blotting. RESULTS: In vitro, a significant, dose dependent suppressive effect of CC10 was found on T(H)2 cytokine expression, but not IFN-gamma, in splenocytes of antigen-sensitized mice. A similar suppressive effect was also noted in polarized CD4(+) T(H)2 cells, but not in naive CD4(+) T cells. In contrast, CC10 was able to induce IFN-gamma expression in naive CD4(+) T cells, but not in polarized T(H)1 cells. Furthermore, the suppression of T(H)2 cytokine expression was concomitant with reduction of a critical transcription factor, GATA-3. Of significance was the finding that although no significant change was found in the decay kinetics of T(H)2 cytokine transcripts, a significant decrease in mRNA stability of GATA-3 was seen in CC10-treated cells. In vivo, reconstitution of the CC10 gene in CC10-deficient mice resulted in significantly lower levels of T(H)2 cytokines, concomitant with a decrease in GATA-3 expression, after challenge with Ag compared with those seen in mock transduced mice, which are associated with reduced levels of pulmonary eosinophilia. CONCLUSION: These results demonstrate, that CC10 plays a direct role in the regulation of T-cell-mediated inflammatory responses. PMID- 15356575 TI - Association study suggests opposite effects of polymorphisms within IL8 on bronchial asthma and respiratory syncytial virus bronchiolitis. AB - BACKGROUND: IL-8 is a strong inductor of inflammation. Accordingly, it plays a pivotal role in acute inflammatory responses during respiratory syncytial virus (RSV) infections and in chronic inflammatory diseases such as bronchial asthma and juvenile idiopathic arthritis. Recently, 2 studies have found association of the polymorphism -251A of IL8 with RSV bronchiolitis. Furthermore, epidemiologic studies have demonstrated an increased risk for the development of asthma after RSV bronchiolitis, and a common genetic background for the 2 diseases is currently being discussed. OBJECTIVE: This study investigated whether IL-8 is in association with asthma and/or arthritis and whether the results can confirm a common genetic background of RSV bronchiolitis and asthma. METHODS: The polymorphisms -A251T, C781T, C1633T, and A2767T within IL8 were genotyped in the following 4 populations: children with asthma, atopic children, children with juvenile idiopathic arthritis, and control subjects. Statistical analysis made use of the Armitage trend test and the software program Arlequine. RESULTS: Association of all polymorphisms was found with asthma ( P =.008 to P =.03). Surprisingly -251T was associated with asthma, which is the opposite allele as described in association with RSV bronchiolitis. Furthermore, all polymorphisms were significantly more common in children with arthritis than in asthmatic children ( P =.006 to P =.02). No association was seen with the diagnosis of arthritis per se or with atopy. CONCLUSION: This is the first study to describe association of IL-8 with asthma and a significant inverse distribution of the polymorphisms in juvenile idiopathic arthritis. In addition, the results of this study might suggest that RSV bronchiolitis and bronchial asthma have at least some different genetic factors. PMID- 15356576 TI - Primary immunodeficiency diseases: an update. AB - Although relatively rare, primary immune deficiency diseases (PIDs) provide an excellent window into the functioning of the immune system. In the late 1960s, observations on these diseases, with their associated infections and genetics, bisected the immune system into humoral immunity and cell-mediated immunity. These diseases also represent a challenge in their diagnosis and treatment. Beginning in 1970, a unified nomenclature for the then-known primary immunodeficiency diseases was created by a committee convened by the World Health Organization. Since then, and later under the aegis of the International Union of Immunological Societies, an international committee of experts has met every 2 to 3 years to update the classification of PIDs. During the past 15 years, the molecular basis of more than 100 PIDs has been elucidated. This update results from the latest meeting of this committee in Sintra, Portugal, June 2003, which followed 2(1/2) days of scientific discussions. PMID- 15356577 TI - Carl Prausnitz: a personal memoir. PMID- 15356578 TI - Prausnitz and Kustner phenomenon: the P-K reaction. PMID- 15356588 TI - Experiments in publishing. PMID- 15356590 TI - Reviewers caution NASA over plans for nuclear-powered craft. PMID- 15356589 TI - Helping depressed children. PMID- 15356591 TI - Peer-reviewed paper defends theory of intelligent design. PMID- 15356592 TI - Conservers plead for funds to protect Europe's heritage. PMID- 15356593 TI - Biomedical agency floats open-access plan. PMID- 15356594 TI - Ocean fix for climate change finds tentative support. PMID- 15356595 TI - China increases share of global scientific publications. PMID- 15356596 TI - Suppliers step in to manage chemical use. PMID- 15356598 TI - Only pride hurt as predicted quake fails to strike California. PMID- 15356597 TI - US chemist attacks consumer magazine's food safety work. PMID- 15356600 TI - Lightning research: the bolt catchers. PMID- 15356601 TI - Antidepressants: bitter pills. PMID- 15356602 TI - Classroom volunteers inspire young ecologists. PMID- 15356603 TI - Science priorities ignore Colombia's water needs. PMID- 15356604 TI - Complexity of the body calls for animal research. PMID- 15356609 TI - Natural proportions. PMID- 15356610 TI - Palaeoclimate: into an ice age. PMID- 15356611 TI - Evolutionary biology: Early evolution comes full circle. PMID- 15356612 TI - Neurobiology: feeding the brain. PMID- 15356613 TI - Condensed-matter physics: the qubit and the cavity. PMID- 15356614 TI - Animal behaviour: relative size in the mating game. PMID- 15356616 TI - Cell biology: an unexpected social servant. PMID- 15356617 TI - Obituary: Edward B. Lewis (1918-2004). PMID- 15356619 TI - Palaeontology: parental care in an ornithischian dinosaur. AB - Crocodilians and birds show extensive parental care of their young, but whether this behaviour evolved independently in these two groups of living archosaurs is unknown - in part because features of parenting among related fossil groups such as dinosaurs are unclear. A dramatic specimen of the small ornithischian dinosaur Psittacosaurus sp. (Dalian Natural History Museum D2156) from Liaoning in China reveals a single adult clustered with 34 juveniles within an area of 0.5 square metres, providing strong evidence for post-hatching parental care in Dinosauria. PMID- 15356620 TI - Bird song: superfast muscles control dove's trill. AB - Bird songs frequently contain trilling sounds that demand extremely fast vocalization control. Here we show that doves control their syrinx, a vocal organ that is unique to birds, by using superfast muscles. These muscles, which are similar to those that operate highly specialist acoustic organs such as the rattle of the rattlesnake, are among the fastest vertebrate muscles known and could be much more widespread than previously thought if they are the principal muscle type used to control bird songs. PMID- 15356621 TI - High-resolution record of Northern Hemisphere climate extending into the last interglacial period. AB - Two deep ice cores from central Greenland, drilled in the 1990s, have played a key role in climate reconstructions of the Northern Hemisphere, but the oldest sections of the cores were disturbed in chronology owing to ice folding near the bedrock. Here we present an undisturbed climate record from a North Greenland ice core, which extends back to 123,000 years before the present, within the last interglacial period. The oxygen isotopes in the ice imply that climate was stable during the last interglacial period, with temperatures 5 degrees C warmer than today. We find unexpectedly large temperature differences between our new record from northern Greenland and the undisturbed sections of the cores from central Greenland, suggesting that the extent of ice in the Northern Hemisphere modulated the latitudinal temperature gradients in Greenland. This record shows a slow decline in temperatures that marked the initiation of the last glacial period. Our record reveals a hitherto unrecognized warm period initiated by an abrupt climate warming about 115,000 years ago, before glacial conditions were fully developed. This event does not appear to have an immediate Antarctic counterpart, suggesting that the climate see-saw between the hemispheres (which dominated the last glacial period) was not operating at this time. PMID- 15356622 TI - The ring of life provides evidence for a genome fusion origin of eukaryotes. AB - Genomes hold within them the record of the evolution of life on Earth. But genome fusions and horizontal gene transfer seem to have obscured sufficiently the gene sequence record such that it is difficult to reconstruct the phylogenetic tree of life. Here we determine the general outline of the tree using complete genome data from representative prokaryotes and eukaryotes and a new genome analysis method that makes it possible to reconstruct ancient genome fusions and phylogenetic trees. Our analyses indicate that the eukaryotic genome resulted from a fusion of two diverse prokaryotic genomes, and therefore at the deepest levels linking prokaryotes and eukaryotes, the tree of life is actually a ring of life. One fusion partner branches from deep within an ancient photosynthetic clade, and the other is related to the archaeal prokaryotes. The eubacterial organism is either a proteobacterium, or a member of a larger photosynthetic clade that includes the Cyanobacteria and the Proteobacteria. PMID- 15356623 TI - Implications for hydrologic processes on Mars from extensive bedrock outcrops throughout Terra Meridiani. AB - Grey haematite was recently detected in the Terra Meridiani region of Mars by the Thermal Emission Spectrometer onboard the Mars Global Surveyor spacecraft. The formation of haematite on Earth often requires liquid water to be present for long periods of time, making this an important target for deciphering the history of water on Mars. The Mars Exploration Rover Opportunity landed in Meridiani early in 2004 and has since discovered light-toned bedrock outcrops rich in chemical and textural signatures of long-term water interaction locally at the landing site. Here I use remote sensing data to show that the light-toned outcrops at the landing site are not a local phenomenon. Instead, they are observable throughout the haematite-bearing plains in both visible and thermal infrared remote sensing data. Moreover, the light-toned material outcrops around much of the margin and is mappable for hundreds of kilometres to the north, east and west of the plains. I infer that 3 x 10(5) km(2) of this material is exposed over 20 degrees of longitude, indicating the extended presence of surface or near surface water over a large region of Mars. PMID- 15356624 TI - Coherent dynamics of a flux qubit coupled to a harmonic oscillator. AB - In the emerging field of quantum computation and quantum information, superconducting devices are promising candidates for the implementation of solid state quantum bits (qubits). Single-qubit operations, direct coupling between two qubits and the realization of a quantum gate have been reported. However, complex manipulation of entangled states-such as the coupling of a two-level system to a quantum harmonic oscillator, as demonstrated in ion/atom-trap experiments and cavity quantum electrodynamics-has yet to be achieved for superconducting devices. Here we demonstrate entanglement between a superconducting flux qubit (a two-level system) and a superconducting quantum interference device (SQUID). The latter provides the measurement system for detecting the quantum states; it is also an effective inductance that, in parallel with an external shunt capacitance, acts as a harmonic oscillator. We achieve generation and control of the entangled state by performing microwave spectroscopy and detecting the resultant Rabi oscillations of the coupled system. PMID- 15356625 TI - Strong coupling of a single photon to a superconducting qubit using circuit quantum electrodynamics. AB - The interaction of matter and light is one of the fundamental processes occurring in nature, and its most elementary form is realized when a single atom interacts with a single photon. Reaching this regime has been a major focus of research in atomic physics and quantum optics for several decades and has generated the field of cavity quantum electrodynamics. Here we perform an experiment in which a superconducting two-level system, playing the role of an artificial atom, is coupled to an on-chip cavity consisting of a superconducting transmission line resonator. We show that the strong coupling regime can be attained in a solid state system, and we experimentally observe the coherent interaction of a superconducting two-level system with a single microwave photon. The concept of circuit quantum electrodynamics opens many new possibilities for studying the strong interaction of light and matter. This system can also be exploited for quantum information processing and quantum communication and may lead to new approaches for single photon generation and detection. PMID- 15356626 TI - Generation of ultraviolet entangled photons in a semiconductor. AB - Entanglement is one of the key features of quantum information and communications technology. The method that has been used most frequently to generate highly entangled pairs of photons is parametric down-conversion. Short-wavelength entangled photons are desirable for generating further entanglement between three or four photons, but it is difficult to use parametric down-conversion to generate suitably energetic entangled photon pairs. One method that is expected to be applicable for the generation of such photons is resonant hyper-parametric scattering (RHPS): a pair of entangled photons is generated in a semiconductor via an electronically resonant third-order nonlinear optical process. Semiconductor-based sources of entangled photons would also be advantageous for practical quantum technologies, but attempts to generate entangled photons in semiconductors have not yet been successful. Here we report experimental evidence for the generation of ultraviolet entangled photon pairs by means of biexciton resonant RHPS in a single crystal of the semiconductor CuCl. We anticipate that our results will open the way to the generation of entangled photons by current injection, analogous to current-driven single photon sources. PMID- 15356627 TI - A high-performance cathode for the next generation of solid-oxide fuel cells. AB - Fuel cells directly and efficiently convert chemical energy to electrical energy. Of the various fuel cell types, solid-oxide fuel cells (SOFCs) combine the benefits of environmentally benign power generation with fuel flexibility. However, the necessity for high operating temperatures (800-1,000 degrees C) has resulted in high costs and materials compatibility challenges. As a consequence, significant effort has been devoted to the development of intermediate temperature (500-700 degrees C) SOFCs. A key obstacle to reduced-temperature operation of SOFCs is the poor activity of traditional cathode materials for electrochemical reduction of oxygen in this temperature regime. Here we present Ba(0.5)Sr(0.5)Co(0.8)Fe(0.2)O(3-delta)(BSCF) as a new cathode material for reduced-temperature SOFC operation. BSCF, incorporated into a thin-film doped ceria fuel cell, exhibits high power densities (1,010 mW cm(-2) and 402 mW cm(-2) at 600 degrees C and 500 degrees C, respectively) when operated with humidified hydrogen as the fuel and air as the cathode gas. We further demonstrate that BSCF is ideally suited to 'single-chamber' fuel-cell operation, where anode and cathode reactions take place within the same physical chamber. The high power output of BSCF cathodes results from the high rate of oxygen diffusion through the material. By enabling operation at reduced temperatures, BSCF cathodes may result in widespread practical implementation of SOFCs. PMID- 15356628 TI - The transition to a sulphidic ocean approximately 1.84 billion years ago. AB - The Proterozoic aeon (2.5 to 0.54 billion years (Gyr) ago) marks the time between the largely anoxic world of the Archean (> 2.5 Gyr ago) and the dominantly oxic world of the Phanerozoic (< 0.54 Gyr ago). The course of ocean chemistry through the Proterozoic has traditionally been explained by progressive oxygenation of the deep ocean in response to an increase in atmospheric oxygen around 2.3 Gyr ago. This postulated rise in the oxygen content of the ocean is in turn thought to have led to the oxidation of dissolved iron, Fe(II), thus ending the deposition of banded iron formations (BIF) around 1.8 Gyr ago. An alternative interpretation suggests that the increasing atmospheric oxygen levels enhanced sulphide weathering on land and the flux of sulphate to the oceans. This increased rates of sulphate reduction, resulting in Fe(II) removal in the form of pyrite as the oceans became sulphidic. Here we investigate sediments from the approximately 1.8-Gyr-old Animikie group, Canada, which were deposited during the final stages of the main global period of BIF deposition. This allows us to evaluate the two competing hypotheses for the termination of BIF deposition. We use iron-sulphur-carbon (Fe-S-C) systematics to demonstrate continued ocean anoxia after the final global deposition of BIF and show that a transition to sulphidic bottom waters was ultimately responsible for the termination of BIF deposition. Sulphidic conditions may have persisted until a second major rise in oxygen between 0.8 to 0.58 Gyr ago, possibly reducing global rates of primary production and arresting the pace of algal evolution. PMID- 15356629 TI - Genetic variation increases during biological invasion by a Cuban lizard. AB - A genetic paradox exists in invasion biology: how do introduced populations, whose genetic variation has probably been depleted by population bottlenecks, persist and adapt to new conditions? Lessons from conservation genetics show that reduced genetic variation due to genetic drift and founder effects limits the ability of a population to adapt, and small population size increases the risk of extinction. Nonetheless, many introduced species experiencing these same conditions during initial introductions persist, expand their ranges, evolve rapidly and become invasive. To address this issue, we studied the brown anole, a worldwide invasive lizard. Genetic analyses indicate that at least eight introductions have occurred in Florida from across this lizard's native range, blending genetic variation from different geographic source populations and producing populations that contain substantially more, not less, genetic variation than native populations. Moreover, recently introduced brown anole populations around the world originate from Florida, and some have maintained these elevated levels of genetic variation. Here we show that one key to invasion success may be the occurrence of multiple introductions that transform among population variation in native ranges to within-population variation in introduced areas. Furthermore, these genetically variable populations may be particularly potent sources for introductions elsewhere. The growing problem of invasive species introductions brings considerable economic and biological costs. If these costs are to be mitigated, a greater understanding of the causes, progression and consequences of biological invasions is needed. PMID- 15356630 TI - Ecosystem stability and compensatory effects in the Inner Mongolia grassland. AB - Numerous studies have suggested that biodiversity reduces variability in ecosystem productivity through compensatory effects; that is, a species increases in its abundance in response to the reduction of another in a fluctuating environment. But this view has been challenged on several grounds. Because most studies have been based on artificially constructed grasslands with short duration, long-term studies of natural ecosystems are needed. On the basis of a 24-year study of the Inner Mongolia grassland, here we present three key findings. First, that January-July precipitation is the primary climatic factor causing fluctuations in community biomass production; second, that ecosystem stability (conversely related to variability in community biomass production) increases progressively along the hierarchy of organizational levels (that is, from species to functional group to whole community); and finally, that the community-level stability seems to arise from compensatory interactions among major components at both species and functional group levels. From a hierarchical perspective, our results corroborate some previous findings of compensatory effects. Undisturbed mature steppe ecosystems seem to culminate with high biodiversity, productivity and ecosystem stability concurrently. Because these relationships are correlational, further studies are necessary to verify the causation among these factors. Our study provides new insights for better management and restoration of the rapidly degrading Inner Mongolia grassland. PMID- 15356631 TI - Recollection-like memory retrieval in rats is dependent on the hippocampus. AB - Recognition memory may be supported by two independent types of retrieval, conscious recollection of a specific experience and a sense of familiarity gained from previous exposure to particular stimuli. In humans, signal detection techniques have been used to distinguish recollection and familiarity, respectively, in asymmetrical and curvilinear components of their receiver operating characteristic (ROC) curves, standard curves that represent item recognition across different levels of confidence or bias. To determine whether animals also employ multiple processes in recognition memory and to explore the anatomical basis of this distinction, we adapted these techniques to examine odour recognition memory in rats. Their ROC curve had asymmetrical and curvilinear components, indicating the existence of both recollection and familiarity in rats. Furthermore, following selective damage to the hippocampus the ROC curve became entirely symmetrical and remained curvilinear, supporting the view that the hippocampus specifically mediates the capacity for recollection. PMID- 15356632 TI - Restricted growth of Schwann cells lacking Cajal bands slows conduction in myelinated nerves. AB - Nerve impulses are propagated at nodes of Ranvier in the myelinated nerves of vertebrates. Internodal distances have been proposed to affect the velocity of nerve impulse conduction; however, direct evidence is lacking, and the cellular mechanisms that might regulate the length of the myelinated segments are unknown. Ramon y Cajal described longitudinal and transverse bands of cytoplasm or trabeculae in internodal Schwann cells and suggested that they had a nutritive function. Here we show that internodal growth in wild-type nerves is precisely matched to nerve extension, but disruption of the cytoplasmic bands in Periaxin null mice impairs Schwann cell elongation during nerve growth. By contrast, myelination proceeds normally. The capacity of wild-type and mutant Schwann cells to elongate is cell-autonomous, indicating that passive stretching can account for the lengthening of the internode during limb growth. As predicted on theoretical grounds, decreased internodal distances strikingly decrease conduction velocities and so affect motor function. We propose that microtubule based transport in the longitudinal bands of Cajal permits internodal Schwann cells to lengthen in response to axonal growth, thus ensuring rapid nerve impulse transmission. PMID- 15356633 TI - Calcium transients in astrocyte endfeet cause cerebrovascular constrictions. AB - Cerebral blood flow (CBF) is coupled to neuronal activity and is imaged in vivo to map brain activation. CBF is also modified by afferent projection fibres that release vasoactive neurotransmitters in the perivascular region, principally on the astrocyte endfeet that outline cerebral blood vessels. However, the role of astrocytes in the regulation of cerebrovascular tone remains uncertain. Here we determine the impact of intracellular Ca(2+) concentrations ([Ca(2+)](i)) in astrocytes on the diameter of small arterioles by using two-photon Ca(2+) uncaging to increase [Ca(2+)](i). Vascular constrictions occurred when Ca(2+) waves evoked by uncaging propagated into the astrocyte endfeet and caused large increases in [Ca(2+)](i). The vasoactive neurotransmitter noradrenaline increased [Ca(2+)](i) in the astrocyte endfeet, the peak of which preceded the onset of arteriole constriction. Depressing increases in astrocyte [Ca(2+)](i) with BAPTA inhibited the vascular constrictions in noradrenaline. We find that constrictions induced in the cerebrovasculature by increased [Ca(2+)](i) in astrocyte endfeet are generated through the phospholipase A(2)-arachidonic acid pathway and 20 hydroxyeicosatetraenoic acid production. Vasoconstriction by astrocytes is a previously unknown mechanism for the regulation of CBF. PMID- 15356634 TI - Cytoplasmic PML function in TGF-beta signalling. AB - Transforming growth factor beta (TGF-beta) is a pluripotent cytokine that controls key tumour suppressive functions, but cancer cells are often unresponsive to it. The promyelocytic leukaemia (PML) tumour suppressor of acute promyelocytic leukaemia (APL) accumulates in the PML nuclear body, but cytoplasmic PML isoforms of unknown function have also been described. Here we show that cytoplasmic Pml is an essential modulator of TGF-beta signalling. Pml null primary cells are resistant to TGF-beta-dependent growth arrest, induction of cellular senescence and apoptosis. These cells also have impaired phosphorylation and nuclear translocation of the TGF-beta signalling proteins Smad2 and Smad3, as well as impaired induction of TGF-beta target genes. Expression of cytoplasmic Pml is induced by TGF-beta. Furthermore, cytoplasmic PML physically interacts with Smad2/3 and SARA (Smad anchor for receptor activation) and is required for association of Smad2/3 with SARA and for the accumulation of SARA and TGF-beta receptor in the early endosome. The PML RARalpha oncoprotein of APL can antagonize cytoplasmic PML function and APL cells have defects in TGF-beta signalling similar to those observed in Pml-null cells. Our findings identify cytoplasmic PML as a critical TGF-beta regulator, and further implicate deregulated TGF-beta signalling in cancer pathogenesis. PMID- 15356635 TI - The joy of the lab. PMID- 15356637 TI - Nuts & bolts. PMID- 15356639 TI - MR-based in vivo hippocampal volumetrics: 2. Findings in neuropsychiatric disorders. AB - Magnetic resonance imaging (MRI) has opened a new window to the brain. Measuring hippocampal volume with MRI has provided important information about several neuropsychiatric disorders. We reviewed the literature and selected all English language, human subject, data-driven papers on hippocampal volumetry, yielding a database of 423 records. Smaller hippocampal volumes have been reported in epilepsy, Alzheimer's disease, dementia, mild cognitive impairment, the aged, traumatic brain injury, cardiac arrest, Parkinson's disease, Huntington's disease, Cushing's disease, herpes simplex encephalitis, Turner's syndrome, Down's syndrome, survivors of low birth weight, schizophrenia, major depression, posttraumatic stress disorder, chronic alcoholism, borderline personality disorder, obsessive-compulsive disorder, and antisocial personality disorder. Significantly larger hippocampal volumes have been correlated with autism and children with fragile X syndrome. Preservation of hippocampal volume has been reported in congenital hyperplasia, children with fetal alcohol syndrome, anorexia nervosa, attention-deficit and hyperactivity disorder, bipolar disorder, and panic disorder. Possible mechanisms of hippocampal volume loss in neuropsychiatric disorders are discussed. PMID- 15356640 TI - V(H) gene analysis of hairy cell leukemia reveals a homogeneous mutation status and suggests its marginal zone B-cell origin. PMID- 15356641 TI - Pretransplant imatinib can improve the outcome of nonmyeloablative stem cell transplantation without increasing the morbidity in Philadelphia chromosome positive chronic myeloid leukemia. PMID- 15356642 TI - Variable frequencies of MALT lymphoma-associated genetic aberrations in MALT lymphomas of different sites. AB - Although several recurrent genetic aberrations are known to occur in MALT lymphoma, no comprehensive study on the most prevalent MALT lymphoma-associated genetic aberrations is available. We therefore screened 252 primary MALT lymphomas for translocations t(11;18)(q21;q21), t(14;18)(q32;q21), and t(1;14)(p22;q32), and trisomies 3 and 18. The above-listed translocations occurred mutually exclusively and were detected overall in 13.5, 10.8, and 1.6% of the cases; trisomy 3 and/or 18 occurred in 42.1%. The frequency at which the translocations occurred varied markedly with the primary site of disease. The t(11;18)(q21;q21) was mainly detected in pulmonary and gastric tumors, whereas the t(14;18)(q32;q21) was most commonly found in lesions of the ocular adnexa/orbit, skin, and salivary glands. Trisomies 3 and 18 each occurred most frequently in intestinal and salivary gland MALT lymphomas. Our results demonstrate that the three translocations and trisomies 3 and 18 occur at markedly variable frequencies in MALT lymphoma of different sites. PMID- 15356643 TI - Altered splicing in hematological malignancies reveals a tissue-specific translational block of the Src-family tyrosine kinase fyn brain isoform expression. PMID- 15356645 TI - Differential involvement of Bax and Bak in TRAIL-mediated apoptosis of leukemic T cells. AB - TRAIL-induced apoptosis has been considered a promising therapeutic approach for tumors that are resistant to chemotherapy, which is usually mediated via mitochondrial apoptotic cascades. Recent studies have shown that in certain cancer cells, TRAIL-mediated apoptosis is also dependent on mitochondrial involvement, suggesting that similar mechanisms of resistance to chemotherapy might be implicated in the resistance of tumor cells to TRAIL. We have used TRAIL resistant leukemic cells that are deficient in both Bax and Bak to determine the roles of these Bcl-2 members in TRAIL-mediated apoptosis. Exposure of these cells to TRAIL did not have an impact on cell viability, although it induced the processing of caspase-3 to its active p20 subunit. The activity of the p20 caspase-3 appeared to be inhibited as no autoprocessing of this p20 subunit or cleavage of known caspase-3 substrates were detected. Also, in the absence of Bax and Bak, no release of mitochondrial apoptogenic proteins was observed following TRAIL treatment. Adenoviral transduction of the Bax, but not the Bak gene, to the Bax/Bak-deficient leukemic cells rendered them TRAIL-sensitive as assessed by enhanced apoptotic death and caspase-3 processing. These findings demonstrate preferential utilization of Bax over Bak in leukemic cell response to specific apoptotic stimulation. PMID- 15356644 TI - Controlling TRAIL-mediated caspase-3 activation. PMID- 15356646 TI - Phase II study of theophylline in chronic lymphocytic leukemia: a study of the Eastern Cooperative Oncology Group (E4998). AB - The Eastern Cooperative Oncology Group (ECOG) performed a phase 2 study in B-cell chronic lymphocytic leukemia (CLL) of oral theophylline, a methylxanthine that inhibits cyclic nucleotide phosphodiesterases, thereby inducing the intracellular accumulation of cyclic adenosine monophosphate (cAMP). In 25 patients with Rai stages 0-I, theophylline, 200 mg given orally every 12 h was well tolerated. There was one complete response after 22.5 months of treatment, which continues at 27+ months, and 18 other patients had stable disease. In vitro exposure of patients' lymphocytes to aminophylline (75-250 microg/ml), the soluble form of theophylline, resulted in dose- and time-dependent induction of apoptosis in 9/20 patients studied. Apoptosis was documented flow-cytometrically by monitoring the expression of bcl-2 and bax, forward light scatter, fluorescence intensity of binding of CD45 antibody, and the binding of annexin. Patients whose leukemic lymphocytes were susceptible to apoptosis induction by aminophylline in vitro experienced a significantly longer progression-free survival than patients whose cells were resistant to the drug in culture (P=0.025). This suggests that in a CLL population treated with theophylline, induction of an apoptotic response to the drug in vitro is prognostic for absence of clinical progression. PMID- 15356647 TI - Riddle: what do aplastic anemia, acute promyelocytic leukemia, and chronic myeloid leukemia have in common? AB - Secondary myelodysplastic syndrome (MDS)/acute leukemia frequently evolves from severe aplastic anemia (SAA) following immunosuppressive therapy. Secondary clonal cytogenetic abnormalities have now been reported after noncytotoxic therapy in two additional settings: all trans retinoic acid (ATRA) treatment of acute promyelocytic leukemia (APL) and imatinib for chronic myeloid leukemia (CML). We propose that SAA, APL, CML, and MDS represent different manifestations of generalized insults to the bone marrow. In SAA, the insult to hematopoietic progenitors leads to an immune attack, while in APL, CML, and MDS, it gives rise to the malignant clones. A primary insult to bone marrow could simultaneously lead to several abnormal hematopoietic cell clones, with one dominating and the others present but below the level of detection. Such a 'field leukemogenic effect' would be analogous to the 'field cancerization effect' described in solid tumors. Nonspecific cytotoxic therapies, including antileukemic chemotherapy and allogeneic transplantation, have broad activity that could inhibit both the overt disease and other undetectable coexistent abnormal clones. In contrast, disease specific targeted therapy such as immunosuppressive therapy in aplastic anemia, ATRA in APL, or imatinib in CML would have no activity against other abnormal clones, allowing them to expand and become detectable as the dominant clone declines. PMID- 15356648 TI - PI3-K/AKT/FKHR and MAPK signaling cascades are redundantly stimulated by a variety of cytokines and contribute independently to proliferation and survival of multiple myeloma cells. AB - IL-6 has been reported to play a central role in growth and survival of multiple myeloma (MM) cells. However, recently we have demonstrated that in the presence of bone marrow stromal cells, survival of MM cells becomes independent of the IL 6/gp130/STAT3 pathway questioning the singular role of IL-6 in MM. Therefore, it was the aim of this study to identify additional factors and signaling pathways that might contribute to the growth and survival of MM cells. We found that in addition to IL-6 a number of bone marrow derived cytokines such as LIF, VEGF, bFGF, MIP-1alpha, SDF-1alpha, IL-1beta, SCF and IL-3 activate the MAPK pathway and induce proliferation of MM.1S and RPMI-8226 MM cells. In addition, these cytokines independently phosphorylate the forkhead family member FKHR via PI3 K/AKT and support survival of primary human MM cells. Inhibition of these pathways induces apoptosis in MM cell lines and primary MM cells. Thus, we provide evidence that in addition to IL-6 a number of different factors trigger important growth-promoting pathways to support the proliferation and survival of MM cells. Therefore, blocking such pathways, rather than blocking a single factor, might be a promising approach for the development of novel treatment strategies in MM. PMID- 15356649 TI - Treatment of children with newly diagnosed acute promyelocytic leukemia with arsenic trioxide: a single center experience. AB - A total of 11 children (five males and six females) with hypergranular type of acute promyelocytic leukemia (APML) were treated with intravenous arsenic trioxide (As(2)O(3)) between December 1998 and October 2003. Eight cycles of As(2)O(3) (0.15 mg/kg/day) were administered (induction, consolidation and six cycles of maintenance) over a period of 12 months. The median WBC count at diagnosis was 3400/mm(3) (range: 800-9800). In all, 10 patients (91%) achieved hematological remission at a mean duration of 48 days (range: 41-60) with all 10 patients achieving molecular remission at a median duration of 81 days (range: 64 109). Toxicity was minimal with leukocytosis in six patients, ichthyosis and hyperpigmentation of skin in five and mild peripheral neuropathy in one patient. One patient who relapsed 6 months after completing therapy achieved a second hematological and molecular remission with As(2)O(3). With a median follow-up of 30 months (range: 4-62), the overall (OS) survival is 91% with a relapse-free survival (RFS) of 81%. As(2)O(3) achieves hematological and molecular remission in majority of newly diagnosed children with APML with minimal toxicity, but long term follow-up is required to evaluate late effects of As(2)O(3) and study the minimum dose and duration required for a sustained remission. PMID- 15356650 TI - Treatment of acute promyelocytic leukemia in children: arsenic or ATRA. PMID- 15356652 TI - Identification of a rare e6a2 BCR-ABL fusion gene during the disease progression of chronic myelomonocytic leukemia: a case report. PMID- 15356653 TI - Clinical significance of minimal residual disease in childhood acute lymphoblastic leukemia after first relapse. PMID- 15356654 TI - Apparent absence of BRCA2 protein in a proportion of acute myeloid leukemia cell lines. PMID- 15356655 TI - Incidence and relevance of secondary chromosome abnormalities in childhood TEL/AML1+ acute lymphoblastic leukemia: an interphase FISH analysis. AB - The aim of the present study was to determine the frequency and clinical relevance of the most common secondary karyotype abnormalities in TEL/AML1+ B cell precursor acute lymphoblastic leukemia (ALL) as assessed with fluorescence in situ hybridization (FISH) analyses. Screening of 372 patients who were enrolled in two consecutive Austrian childhood ALL multicenter trials identified 94 (25%) TEL/AML1+ cases. TEL deletions, trisomy 21 and an additional der(21)t(12;21) were detected in 52 (55%), 13 (14%) and 14 (15%) TEL/AML1+ patients, respectively. The 12p aberrations (P=0.001) and near tetraploidy (P=0.045) were more common in TEL/AML1+ patients, whereas the incidence of diploidy, pseudodiploidy, hypodiploidy, low hyperdiploidy, near triploidy, del(6q), chromosome 9 and 11q23 abnormalities was similar among TEL/AML1+ and TEL/AML1- patients. None of the TEL/AML1+ patients had a high hyperdiploid karyotype. Univariate analysis indicated that among TEL/AML1+ patients those with a deletion of the nontranslocated TEL allele had a worse prognosis than those without this abnormality (P=0.034). We concluded that the type and incidence of the most common secondary aberrations in TEL/AML1+ ALL can be conveniently identified with little additional effort during interphase screening with appropriate TEL and AML1 FISH probes. We also provided preliminary evidence that the deletion of the nontranslocated TEL allele may adversely influence the clinical course of TEL/AML1+ ALL. PMID- 15356656 TI - Farnesyltransferase inhibitor BMS-214662 induces apoptosis in B-cell chronic lymphocytic leukemia cells. AB - B-cell chronic lymphocytic leukemia (B-CLL) cells develop resistance to nucleoside analogs over time. This chemoresistance may be caused by selection for B-CLL cells with defects in the particular apoptosis pathway triggered by these drugs. Therefore, anticancer agents that induce apoptosis through alternative pathways might be useful in treating chemoresistant B-CLL. Farnesyltransferase inhibitors (FTIs) are a class of synthetic drugs with definite molecular targets, which have demonstrated cytotoxicity against leukemic cell lines. We have studied the ex vivo effect of the FTI BMS-214662 on cells from 18 patients with B-CLL. Low concentrations (<1 microM) of BMS-214662 prevented farnesylation of the chaperone marker HDJ-2 and had no effect on Akt activation. BMS-214662 induced apoptosis in B-CLL cells from all patients studied, including those showing resistance to cladribine and fludarabine ex vivo and in vivo. Treatment with BMS 214662 induced loss of mitochondrial membrane potential (DeltaPsi(m)), phosphatidylserine exposure, proapoptotic conformational changes of Bax and Bak, reduction in Mcl-1 levels and activation of caspases 9 and 3. The general caspase inhibitor Z-VAD-fmk did not prevent BMS-214662-induced cell death. These results indicate that BMS-214662 may be a useful drug for treating B-CLL and, in particular, an alternative for the therapy of purine analog-resistant or relapsed B-CLL. PMID- 15356657 TI - Low-dose oral etoposide-based induction regimen for children with acute lymphoblastic leukemia in first bone marrow relapse. AB - We evaluated the clinical response to low-dose etoposide in relapsed acute lymphoblastic leukemia (ALL). Of the 45 patients with ALL in first bone marrow relapse enrolled on the ALL R15 protocol, 44 had received epipodophyllotoxins during frontline therapy. In the first week of remission induction therapy, patients received etoposide (50 mg/m(2) per day) administered orally as a single agent once or twice daily. On Day 8, patients started to receive dexamethasone, vincristine, and L-asparaginase. Etoposide was administered until Day 22. Two courses of consolidation therapy were followed by continuation therapy or hematopoietic stem cell transplantation. After 7 days of single-agent etoposide treatment, peripheral blast cell counts (P=0.013) and percentages of bone marrow blasts (P=0.016) were significantly reduced. In all, 38 (84.4%) attained second remission. Only time to relapse was significantly associated with outcome (P=0.025): the 5-year event-free survival estimates (+/-se) were 52.0+/-9.6% for those with late relapse and 20.0+/-8.0% for those with early relapse. We conclude that low-dose etoposide administered orally has a cytoreductive effect in relapsed ALL. PMID- 15356658 TI - Malignant hematopoietic cell lines: in vitro models for the study of anaplastic large-cell lymphoma. PMID- 15356659 TI - Establishment of a novel anaplastic large-cell lymphoma-cell line (COST) from a 'small-cell variant' of ALCL. AB - Anaplastic large-cell lymphoma (ALCL) is a distinct biological and cytogenetic entity with a broad spectrum of morphological features (common type, small-cell variant and lymphohistiocytic variant). Few cell lines of ALCL are available and they all originate from primary tumors demonstrating the common type morphology (ie large-sized lymphoma cells). We established a new ALCL cell line (COST) from the peripheral blood of a patient with a small-cell variant of ALCL, at diagnosis. Cells growing in vitro and in SCID mice consisted of two populations, that is, small- and large-sized cells as seen in the patient's tumor. Both large and small malignant cells were positive for CD43/MT1 T-cell associated antigen, perforin, granzyme B and TIA-1, but negative for CD2, CD3, CD5, CD7, CD4 and CD8 antigens. Standard cytogenetic studies as well as multiplex FISH confirmed the presence of the canonical t(2;5)(p23;q35) translocation, but also revealed additional numerical and structural abnormalities. The COST cell line is the first ALCL small-cell variant cell line, and thus provides a potentially useful tool for further functional and molecular studies that should improve our understanding of the small-cell variant of ALCL, which is more frequently complicated by a leukemic phase. PMID- 15356660 TI - Prenatal origin of separate evolution of leukemia in identical twins. AB - Several studies involving identical twins with concordant leukemia and retrospective scrutiny of archived neonatal blood spots have shown that the TEL AML1 fusion gene in childhood acute lymphoblastic leukemia (ALL) frequently arises before birth. A prenatal origin of childhood leukemia was further supported by the detection of clonotypic immunoglobulin gene rearrangements on neonatal blood spots of children with various other subtypes of ALL. However, no comprehensive study is available linking these clonotypic events. We describe a pair of 5-year-old monozygotic twins with concordant TEL-AML1-positive ALL. Separate leukemic clones were identified in the diagnostic samples since distinct IGH and IGK-Kde gene rearrangements could be detected. Additional differences characterizing the leukemic clones included an aberration of the second, nonrearranged TEL allele observed in one twin only. Interestingly, both the identical TEL-AML1 fusion sequence and distinct immunoglobulin gene rearrangements were identified on the neonatal blood spots indicating that separate preleukemic clones evolved already before birth. Finally, we compared the reported twins with an additional 31 children with ALL by using the microarray technology. Gene expression profiling provided evidence that leukemia in twins harbours the same subtype-typical feature as TEL-AML1-positive leukemia in singletons suggesting that the leukemogenesis model might also be applicable generally. PMID- 15356661 TI - The body mass index is a less-sensitive tool for detecting cases with obesity associated co-morbidities in short stature subjects. AB - OBJECTIVE: To assess the ability of the body mass index (BMI) to detect obesity associated morbidity in subjects with a normal or short stature. METHODS: Information was obtained on 119 975 subjects from a cardiovascular risk factors detection program. Standardized questionnaires were used. Capillary glucose and cholesterol concentrations were measured. Diabetes, arterial hypertension and hypercholesterolemia were selected as end points. Sensitivity, specificity and the likelihood ratio for several BMI thresholds were calculated. ROC curves were constructed to identify the BMI cutoff points with best diagnostic performance. The area under the curve (AUC) was used to assess the proficiency of BMI. RESULTS: Short stature (height 25 kg/m(2)), age>18 and <70 y and ability to operate electronic microdevices. INTERVENTIONS: All patients in the control group (n=77) received standard hospital care. Patients in the intervention group (n=45), additionally, measured three times a week, for 6 months, their blood pressure and body weight and transmitted them to an automated call center. These values were not shared with the patients' physician or dietician. MAIN OUTCOME MEASURES: Clinical (body weight, BMI, blood pressure), laboratory (fasting plasma glucose, triglycerides, HDL-cholesterol, total cholesterol) and quality of life parameters (SF-36((R)), Visual Analog Scale of European Quality-5 Dimensions, Obesity Assessment Survey). Data were analyzed in an intention-to-treat-way (last observation carried forward). RESULTS: Drop-out rate was similar in the control and intervention groups: 12 vs 11 percent, respectively, P=NS. There were no significant differences at baseline between intervention and control groups in all main outcome parameters. There were significant decreases for patients in the intervention group in body weight (from 101.6+/-22.4 to 89.2+/-14.7 kg, P=0.002, P=0.05 vs controls at 6 months), total cholesterol (from 247.6+/-42.0 to 220.7+/ 42.6 mg/dl, P=0.002, P=0.05 vs controls at 6 months) and triglycerides (from 148.4+/-35.0 to 122.3+/-31.4 mg/dl, P=0.001, P=0.01 vs controls at 6 months). Intervention group patients made a total of 1997 phone contacts. The number of phone contacts was correlated positively with Social Functioning (SF), Vitality (VT) and Mental Health (MH) scores of SF-36((R)) at baseline (r=0.48, r=0.41, r=0.41, respectively, P=0.05) but not with weight loss. CONCLUSIONS: Home centered, intense treatment through the use of telemedicine can be effective in improving short-term obesity outcomes. PMID- 15356665 TI - Effects of weight gain on medical care costs. AB - OBJECTIVE: To examine in middle-aged adults the effect of medical care costs of large, rapid weight gain compared to weight maintenance. DESIGN: : Retrospective cohort study for a 3-y time period. SETTING AND PARTICIPANTS: Population-based sample (N=15174) of men and women members of a large managed care organization, aged 35-65 y, with a body mass index (BMI) >25 kg/m(2) at baseline. Health-care utilization and costs were measured at baseline and over the 3-y follow-up period. RESULTS: Mean age at baseline was 49.7 y and mean BMI was 31.5 kg/m(2). During the 3-y follow-up period, 40.8% were classified as weight maintainers (+/ 4 pounds), 45.3% gained 5-19 pounds, and 13.9% gained >/=20 pounds. A weight gain of >/=20 pounds was significantly associated with increased total medical care costs in all subgroups evaluated. Among all subjects, for those who gained >/=20 pounds compared to those who maintained weight, the adjusted 3-y increase in costs was 561 dollars. Among the subgroup with baseline comorbidities, the adjusted 3-y change in total medical care costs was 711 dollars. Multivariate analyses showed no significant differences between those who gained 5-19 pounds and those who maintained weight. Baseline BMI and comorbidities were also significant predictors of change in medical care costs, independent of weight gain. CONCLUSION: A large 3-y weight gain (>/=20 lb) in middle-aged overweight and obese adults is associated with a correspondingly larger increase in total medical care costs compared to weight maintainers. The prevention of large weight gains holds promise for significantly reducing future medical care costs. Future studies should examine the causes of rapid weight gain and evaluate approaches to prevent and reverse such weight gain. PMID- 15356666 TI - Overweight and obesity and the burden of disease and disability in elderly men. AB - OBJECTIVE: To examine the prevalence of disease burden and disability associated with overweight and obesity in men aged 60-79 y and to assess whether the current WHO weight guidelines are appropriate in the elderly. DESIGN: Cross-sectional survey 20 y after enrollment. SETTING: General practices in 24 British towns. PARTICIPANTS: In total, 4232 men aged 60-79 y (77% of survivors) with measured weight and height. MAIN OUTCOME MEASURES: Cardiovascular (CV) risk factors, prevalence of diabetes, cardiovascular disease, cancer, disability and regular medication. RESULTS: In total, 17% of the men were obese (body mass index (BMI) >/=30 kg/m(2)) and a further 52% were overweight (BMI 25-29.9 kg/m(2)). Prevalence of hypertension, low HDL-cholesterol, high triglycerides and insulin resistance and the prevalence of most disease outcomes increased with increasing degrees of overweight/obesity. Men in the normal weight range (18.5-24.9 kg/m(2)) had the lowest prevalence of ill health. Compared with normal weight men, obese men showed a two-fold risk of major CVD (odds ratio (OR)=1.96, 95% CI 1.44-2.67) and locomotor disability (OR=2.26, 95% CI 1.66, 3.09) and were nearly three times as likely to have diabetes, CV interventions or to be on CV medication. Over 60% of the prevalence of high insulin resistance was attributable to overweight and obesity as was over a third of diabetes and hypertension, a quarter of locomotor disability and a fifth of major CVD. CONCLUSION: In elderly men, overweight and obesity are associated with a significantly increased burden of disease, in particular CV-related disorders and disability. The current guidelines for overweight and obesity appear to be appropriate in elderly men. PMID- 15356667 TI - Evaluation of adipocyte apoptosis by laser scanning cytometry. AB - OBJECTIVE: Adipocyte apoptosis plays an important role in adipose tissue homeostasis and can be altered under a variety of physiological and pathological conditions. This study was carried out to determine whether laser scanning cytometry (LSC) can be used to measure changes in apoptosis of adipocytes over time. DESIGN: LSC was used to investigate adipocyte apoptosis induced by tumor necrosis factor-alpha (TNF-alpha), a cytokine that is associated with obesity and insulin resistance. LSC, a slide-based solid phase cytofluorometer, provides quantitative flow fluorescence data together with morphological information for apoptotic detection. Both 3T3-L1 cells and rat adipocytes from primary cell culture were incubated with 0 or 25 nM TNF-alpha for up to 24 h. Both the FITC conjugated annexin V/propidium iodide assay and the TUNEL assay were used to distinguish cells with apoptotic characteristics from nonapoptotic cells. RESULTS: Apoptosis did not increase over time in the absence of TNF-alpha for both 3T3-L1 cells and rat primary adipocytes. For both 3T3-L1 cells and rat primary adipocytes, a significant increase in the percentage of apoptotic cells was observed by 3-4 h incubation with TNF-alpha (P<0.05). By 24 h, more than 50% of cells incubated with TNF-alpha were apoptotic (P<0.001). This process was also associated with morphological changes typical of adipocytes undergoing apoptosis. By estimating the percentage of cell subpopulations after different times of incubation with TNF-alpha, we were able to develop grading parameters, based on the adipose apoptotic measurements. CONCLUSION: With morphological information, LSC can be a useful tool to evaluate adipocyte apoptosis. PMID- 15356668 TI - Obesity and impaired fibrinolysis: role of adipose production of plasminogen activator inhibitor-1. AB - Obesity is the central promoter of the metabolic syndrome which also includes disturbed fibrinolysis in addition to hypertension, dyslipidaemia and impaired glucose tolerance/type 2 diabetes mellitus. Plasminogen activator inhibitor-1 (PAI-1) is the most important endogenous inhibitor of tissue plasminogen activator and uro-plasminogen activator, and is a main determinant of fibrinolytic activity. There is now compelling evidence that obesity and, in particular, an abdominal type of body fat distribution are associated with elevated PAI-1 antigen and activity levels. Recent studies established that PAI-1 is expressed in adipose tissue. The greater the fat cell size and the adipose tissue mass, the greater is the contribution of adipose production to circulating PAI-1. Experimental data show that visceral adipose tissue has a higher capacity to produce PAI-1 than subcutaneous adipose tissue. Studies in human adipocytes indicate that PAI-1 synthesis is upregulated by insulin, glucocorticoids, angiotensin II, some fatty acids and, most potently, by cytokines such as tumour necrosis factor-alpha and transforming growth factor-beta, whereas catecholamines reduce PAI-1 production. Interestingly, pharmacological agents such as thiazolidinediones, metformin and AT(1)-receptor antagonists were found to reduce adipose expression of PAI-1. In addition, weight loss by dietary restriction or comprehensive lifestyle modification is effective in lowering PAI-1 plasma levels. In conclusion, impaired fibrinolysis in obesity is probably also due to an increased expression of PAI-1 in adipose tissue. An altered function of the endocrine system and an impaired auto-/paracrine function at the fat cell levels may mediate this disturbance of the fibrinolytic system and thereby increase the risk for cardiovascular disease.. PMID- 15356669 TI - Midarm computerized tomography fat, muscle and total areas correlation with nutritional assessment data. AB - BACKGROUND AND OBJECTIVES: Correlated nutritional assessment data (anthropometric, bioimpedance and biochemical) with computerized tomography (CT) of total, muscle and fat midarm areas. Total body fat and fat-free mass were estimated using bioimpedance. Daily urinary urea and creatinine were also quantified. In all, 28 subjects (13 males, 15 females) were evaluated and, they were clinically divided in obese, malnourished and control subjects. DESIGN: Quantification of total, fat muscle midarm areas by tomography and anthropometry and total body fat and free-fat mass by bioimpedance. RESULTS: CT values were 29% higher for fat area and 4-5% lower for total and muscle midarm areas compared against anthropometric data. The midarm skinfold thickness highly correlated with CT fat midarm area. Total body fat and free-fat mass bioimpedance data showed significant correlation with CT midarm data. Urinary creatinine correlated with CT muscle midarm area. CONCLUSION: Utilization of anthropometry can lead to error estimation of fat and fat-free arm areas and that bioimpedance gives fair correlation between total body and CT midarm measurements. PMID- 15356670 TI - A randomized double-blind placebo-controlled clinical trial of a product containing ephedrine, caffeine, and other ingredients from herbal sources for treatment of overweight and obesity in the absence of lifestyle treatment. AB - OBJECTIVE: To evaluate the efficacy and side effects of an herbal formulation to promote weight loss, as compared to placebo. DESIGN: 12-week multicenter double blind, placebo-controlled, randomized parallel groups design. Study conducted at three clinical sites in New York State. Subjects were randomized to receive either the 'active' product or a 'placebo' supplement for 12 weeks. Minimal steps were taken to influence lifestyle changes with regard to diet or exercise. SUBJECTS: 102 overweight/obese (300.05). CONCLUSION: Results demonstrated the importance of placebo-controlled trials and quantitative outcome measures for the evaluation of 4-AP aimed to enhance gait for chronic, ambulatory SCI persons. Energy expenditure measures and mood may relate more to subjective comments and is suggested for future investigations. PMID- 15356677 TI - Movement patterns and muscular demands during posterior transfers toward an elevated surface in individuals with spinal cord injury. AB - STUDY DESIGN: Three-dimensional kinematic analysis and surface electromyography (EMG) of 10 male adults with complete spinal cord injury (C7 to L2). OBJECTIVE: To examine movement patterns and muscular demands in individuals with spinal cord injury (SCI) during posterior transfers. SETTING: Pathokinesiology Laboratory at a Rehabilitation Centre, Montreal, Canada. METHODS: Kinematic variables that described the positions and angular displacements of the head, trunk, shoulder and elbow were obtained by videotaping markers placed on the subject segments. EMG data were recorded for the biceps, triceps, anterior deltoid, pectoralis major, latissimus dorsi and trapezius muscles of the dominant upper extremity during posterior transfers using surface electrodes. To quantify the muscular demand, the EMG data recorded during the transfers were normalized to values obtained during maximal static contractions (EMGmax). The mean muscular demand was calculated for every muscle during the lift phase of the transfers. The lift phase was determined by pressure-sensitive contacts. RESULTS: All subjects were able to execute the posterior transfers on an even surface, whereas nine subjects completed at least one of the transfers to the elevated surface. A forward flexion pattern at the head and trunk was observed when either one or two hands remained on the lower surface, whereas a lift strategy was seen when both hands were placed on the elevated surface. Transferring to the elevated surface with hands on the lower surface required inferior electromyographic muscular utilization ratio (EMUR) than the transfer on the even surface for all muscles. The lowest EMUR were calculated for the transfer to the elevated surface with hands on the lower surface (triceps (18%), pectoralis major (53.8%), trapezius (66%) and latissimus dorsi (24.5%)) while performing the same transfer with hands on the elevated surface generated the highest EMUR (triceps (40.2%), anterior deltoid (73.2%), trapezius (83.6%) and latissimus dorsi (55.3%)). CONCLUSIONS: Subjects presented different movement characteristics and muscular demands during the posterior transfers. It is suggested that the forward-flexion pattern improves the dynamic trunk stability and reduces the muscular demand required to transfer. High muscular demand developed when hands were positioned on the elevated surface might be due to increased postural control demands on the upper limb and reduced angular momentum. PMID- 15356678 TI - Prostate cancer: palliative care and pain relief. AB - Metastatic prostate cancer is incurable, and both the disease and other comorbidities as well as side effects from treatments can be very disabling for patients who are generally elderly. Past reviews on palliative care in prostate cancer have tended to concentrate on methods of active intervention with palliative intent rather than the principles of palliative care in a broader context. This paper is intended to introduce the principles and practice of modern palliative care as applied to care of patients with prostate cancer. Current National Institute for Clinical Excellence guidelines for urological cancers recommends provision of palliative care for all patients with prostate cancer, according to need. Palliative care aims to improve the quality of life of patients and their families. It is a holistic and personal approach to patient care, consisting of patient centred multiprofessional teams. Symptoms such as pain, psychological, spiritual and social problems are given equal weighting and tackled simultaneously. When treating pain, hormonal manipulation, radiotherapy, chemotherapy and surgery should always be considered alongside analgesic therapy, with realistic aims in mind. Towards the end of life, palliative care for patients will generally increase in line with increasing symptoms and other problems. Achieving a peaceful and comfortable death for our patients should be a priority in hospitals. PMID- 15356679 TI - Type IV collagenase (matrix metalloproteinase-2 and -9) in prostate cancer. AB - BACKGROUND: The type IV collagenases/gelatinases matrix metalloproteinase-2 (MMP 2) and -9 (MMP-9) play an important role in cancer invasion and metastasis. In the present study, we measured the expression of mRNAs and enzymatic activities of MMP-9 and -2 in prostate tissues and serum samples from men with or without prostate cancer. METHODS: A total of 44 tissue samples (three from healthy volunteers, 21 from patients with benign prostate hyperplasia, 10 from patients with localized prostate cancer and 10 from patients with metastatic disease) and 71 serum samples were collected (20 from healthy volunteers, 26 from patients with benign prostatic hyperplasia, 10 from patients with localized cancer, 15 from patients with metastatic cancer). The level of mRNA for MMP-2 and -9 was determined by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The enzymatic activity of MMPs was determined by zymography. RESULTS: Expression of MMP-9 mRNA was significantly higher in malignant than in nonmalignant prostate tissues (P < 0.001), while no significant difference of MMP 2 expression was detected in different prostate tissues. Results of zymography showed that there was significant difference in the enzymatic activity of MMP-9, but not MMP-2, among normal prostate, BPH, localized and metastatic prostate cancer tissues, serum samples (P < 0.05). The active form of MMP-2, with a molecular mass of 62 kDa, was detected in normal prostate, BPH and prostate cancer tissues, but not in the serum samples. Moreover, there was a significant difference in the ratio of the active form (62 kDa) and proform (72 kDa) of MMP-2 among normal, BPH and prostate cancer tissues. This ratio was further increased in metastatic prostate cancer tissues. CONCLUSION: The activity of MMP-9 and the ratio of active form/proform of MMP-2 are associated with the progression and metastasis of prostate cancer. PMID- 15356681 TI - [Short and good information about drugs]. PMID- 15356680 TI - Comparison of external radiation therapy vs radical prostatectomy in lymph node positive prostate cancer patients. AB - INTRODUCTION: Treatment options for lymph node positive prostate cancer are limited. We retrospectively compared patients who underwent external radiotherapy (ERT) to patients treated by radical prostatectomy (RPX). MATERIALS AND METHODS: A total of 102 lymph node positive patients from the RPX series at Ulm University were evaluated. In all, 76 patients received adjuvant androgen withdrawal as part of their primary treatment. In the ERT group, 44 patients were treated at the University of Michigan using a fractionated regimen. Of these, 21 patients received early adjuvant hormonal therapy. Patients with neoadjuvant therapy before RPX or ERT were excluded. RESULTS: In the RPX group, PSA nadir (nadir < or = 0.2 vs > 0.2 ng/ml) showed a strong association with outcome. In the ERT group, pretreatment PSA was an independent predictor of outcome (P = 0.04) and patients with adjuvant hormonal therapy had a significant longer recurrence-free interval compared to patients without adjuvant therapy (P = 0.004). Comparing only patients with adjuvant hormonal treatment after cancer-specific therapy, the ERT treated patients had a borderline longer PSA recurrence-free survival time compared to the RPX-treated patients (P = 0.05). CONCLUSIONS: In case of positive lymph nodes, RPX and ERT might be considered and need to be explained to the patient. For future treatment decisions, the presented findings and a potential survival benefit need to be evaluated in a larger prospective setting. PMID- 15356682 TI - [Is mammography screening beneficial?]. PMID- 15356683 TI - [Information about mammography screening]. PMID- 15356684 TI - [What does influence the prescription of antibiotics to children?]. PMID- 15356685 TI - [Parents' attitudes to the prescription of antibiotics to children]. AB - BACKGROUND: Most children in day care get infections and are prescribed antibiotics. We studied parents' attitude towards such prescription. MATERIAL AND METHODS: Parents (563 out of 1126 surveyed) in 22 day care centres returned a questionnaire on factors in their own situation and in their day care centre with impact on the use of antibiotics. RESULTS: Parents above 30 years of age, well educated and knowledgeable about antibiotics were the most sceptical of such treatment. A negative attitude on the part of employers and colleagues towards absence from work led to more use of antibiotics (p < 0.01). Children in single parent homes were given more antibiotics than those in two-parent homes (p < 0.05). Help from relatives led to less use of antibiotics (p < 0.01). Sick children went to day care in 16% of cases; 57% of those taking antibiotics still went to day care. INTERPRETATION: Parents' situation may influence the use of antibiotics in children. Too many children are taking antibiotics and still go to day care. Support in the workplace, from relatives and from friends may give parents a better chance of staying at home with a sick child, thus preventing the spread of infections and cut down on the use of antibiotics. PMID- 15356686 TI - [The genetics of movement disorders--spinocerebellar degenerations]. AB - BACKGROUND: Hereditary movement disorders include spinocerebellar disorders, a large and heterogeneous group of syndromes with ataxia or spasticity as the prominent symptom. In spite of the vast clinical and genetic heterogeneity, patterns of pathogenesis slowly emerge and help us understand these disorders. MATERIAL AND METHODS: This review is based on personal experience and recent literature. RESULTS: More than 20 types of hereditary spastic paraparesis have been reported. Dominant SPG4 and SPG3 with mutations in the spastin or the atlastin gene have been identified in many countries. The most prevalent type of recessive ataxia in Europe, Friedreich's ataxia, has become a model of integrated clinical-molecular-therapeutic research. More recessive ataxias (AOA1-2) have been described recently. More than 20 autosomal dominant ataxias have been reported, with 12 identified genes including the episodic ataxias, and 9 mapped. SCA7 appears to be the most frequent type in some Nordic countries. INTERPRETATION: A striking feature of many of these diseases is the involvement of very different genes for similar phenotypes. Conversely, very heterogeneous phenotypes are due to single-gene defects. Recently there has been considerable progress in the clinical description of movement disorders and the understanding of their genetic basis. Possible therapies are emerging. PMID- 15356687 TI - [Genetics in movement disorders--dystonia, tremor and chorea]. AB - BACKGROUND: Several dystonias, Huntington's disease, essential tremor and various rare conditions are among the hereditary movement disorders, disorders with considerable genetic and clinical heterogeneity. It may be hoped that better understanding of the genetics and pathogenetic mechanisms involved will improve management. MATERIAL AND METHODS: This review is based on personal experience and recent literature. RESULTS: At least 13 types of dystonias are genetically defined. DYT 1 seems to be the most important, and is easily tested. The Huntington gene is readily available for symptomatic as well as presymptomatic testing. Presymptomatic testing gives rise to ethical concerns and require thorough genetic counselling. Also essential tremor seems to show an autosomal dominant inheritance, but the responsible gene(s) has not been identified. INTERPRETATION: As for other movement disorders, involvement of different genes may give similar phenotypes and defects in a single gene give rise to various different phenotypes. PMID- 15356688 TI - [Overdiagnosis in mammography screening]. AB - BACKGROUND: In Norway and Sweden, the introduction of mammography screening programmes has been associated with about a 50% increase in breast cancer incidence for the screened age groups and almost stable incidence in higher age groups. This suggests that mammography screening results in a substantial degree of overdiagnosis. MATERIAL AND METHODS: In a prospective cohort study of the Norwegian mammography programme we recorded the incidence of breast cancer among women aged 50 to 69 years offered mammography screening as well as the incidence of interval cancer among those attending, and breast cancer among those not attending. We used these data to calculate the incidence of cancer detected by screening among those screened. RESULTS: The incidence of invasive cancer at second and third screening is 2.4 times higher than expected. We estimated that only 42% of the screening-detected invasive cancers would have developed into clinical disease in the absence of a screening programme. When ductal carcinoma in situ is added, only 34% of the screening-detected cancers would have developed into clinical disease. INTERPRETATION: Two thirds of breast cancers detected by screening are overdiagnosed. PMID- 15356689 TI - [Parents' attitude to physician's role in the prescription of antibiotics to their children]. AB - BACKGROUND: Most children in day care get infections and are given antibiotics. In physicians' view, the main reason for excessive use is pressure exerted on them by anxious parents. We studied parents' view of physicians' influence on the use of antibiotics. MATERIAL AND METHOD: 563 parents (50%) in 22 day care centers returned a questionnaire about their children's infections, use of antibiotics, and their experience with the doctors treating their child. RESULTS: 70% of parents had confidence in physicians' decisions. Confidence was significantly higher in doctors that they regularly consulted than in others (p = 0.001). 33% had moved on to a new physician on account of too much use of antibiotics (p < 0.05); these parents' children had received more antibiotics than other children (p < 0.05). Parents were dissatisfied with the time set aside for the consultation and with doctors' information and follow up. They expected advice and guidance, not necessarily a prescription. 47% thought that too much antibiotics are prescribed. Satisfaction was associated with less antibiotics for their own child (p < 0.001). Some parents regarded the prescription of antibiotics as doctors' way of saving time and bringing the consultation to an end. INTERPRETATION: Parents are sceptical of the use of antibiotics in children. More time set aside for the consultation is significantly associated with less use of antibiotics. Better doctor-parent communication may reduce excessive use. PMID- 15356690 TI - [Treatment of acute otitis media in children]. PMID- 15356691 TI - [Eating disorders and personality disorders--possible interactions and their therapeutic implications]. AB - Personality disorders are defined by a characteristic and enduring pattern of behaviour and inner experience that deviates distinctly from culturally defined norms. Examples include social insecurity or even distrust leading to withdrawal, or patterns of impulsiveness, affective instability or, possibly, self-harm. Personality disorders are distinct from other psychiatric disorders primarily by their enduring character. The prevalence of personality disorders among patients with eating disorders depends on population characteristics. In a specialised outpatient clinic that serves a defined catchment area, nearly one third of patients will fulfill the diagnostic criteria for a personality disorder. Hospitalized patients or patients in more specialised units will surely show a considerably higher prevalence. Several modes of interaction between eating disorders and personality disorders could be hypothesized. The treatment of patients with eating disorder should take into account the subgroup with comorbid personality pathology and evaluation and treatment should be planned accordingly, though this requires time and expertise. The issue should also be acknowledged in primary health care and attention be paid to symptoms of eating disorder also in cases in which symptoms of a personality disorder are more pronounced. PMID- 15356692 TI - [Eating disorder and the family--a review]. AB - The importance of the family in eating disorders has been the subject of a great deal of speculation ever since anorexia nervosa was first described some 130 years ago. Given the importance of the family in child and adolescent development, it also has great bearing on how young people learn to deal with food. But the research is inconclusive as to the family's precise role in the development of an eating disorder. There is no support in the literature on which to base a comprehensive understanding of the family setting in which a member develops an eating disorder; this also applies to anorexia nervosa cases. When groups of families afflicted with different psychiatric disorders are compared, it seems that families in which bulimia nervosa occurs tend to be more dysfunctional than families afflicted with anorexia nervosa. It also seems that families afflicted with anorexia nervosa function better than families afflicted by other psychiatric disorders. In this article the research on family functioning in relation to anorexia nervosa and bulimia nervosa is reviewed and its relevance discussed. PMID- 15356693 TI - [Eating disorders--how to work with the family?]. AB - Working with families with a child or an adult with an eating disorder is to work with the resources and limitations in the families, how they could cope with this challenge. Family interventions in eating disorders are counselling, working with the family and family-oriented therapy. Studies show documented effects, especially for patients below 18 with anorexia nervosa who have had this disorder for less than three years. Indications for going from working with the family to family therapy are conflicts between parents or between the child and parents, delayed or disturbed psychological development of the child, or when siblings or other family members are believed to have an effect on the healing. Family therapy is to work with the family as a system. The professional responsibilities, the practical tasks and the goals are about interaction in the family, about relations and communication patterns. Interventions in the family are mainly rooted in systemic family therapy. Motivational methods are important, as are cognitive methods and psychoeducational methods, information and learning how to cope with eating disorders. PMID- 15356694 TI - [Erroneous and unsatisfactory filling in of drug charts--a potential source of medication error]. AB - BACKGROUND: Medication errors may have serious consequences for patients and erroneous drug charts represent one of several causes of error. As part of a quality assurance survey, we studied whether user documentation on the charts was in accordance with the guidelines for chart keeping. MATERIAL AND METHODS: On the same day, the drug charts of all hospitalized patients (n = 401) were copied and a representative sample of 207 charts was reviewed. RESULTS: 15 charts had no prescriptions; 14 of the remaining 192 charts (7 %) were correctly filled in. 152 charts (79 %) had deviations that we considered might pose only minor potential health risk; 26 (14 %) had deviations implying potential risk. No charts had deviations considered to pose major risk. INTERPRETATION: Several deviations from the guidelines were found and better training is needed; we recommend standardisation of drug charts and guidelines. PMID- 15356695 TI - [Prioritization is more than to say no]. PMID- 15356696 TI - [Does insemination with non-anonymous sperm donor have a future?]. PMID- 15356699 TI - [Replay about fertilized embryo research]. PMID- 15356700 TI - [Prevention of neural tube defects with folic acid]. PMID- 15356701 TI - [Evolution, diet and life style]. PMID- 15356703 TI - [Patient rights legislation and the president]. PMID- 15356704 TI - [A new delirium severity scale]. PMID- 15356705 TI - [To help seldom means the same as to please]. PMID- 15356707 TI - [Statins and hypertension]. PMID- 15356709 TI - [Thiazides (only) and hard endpoints]. PMID- 15356710 TI - Tattoo, human identity and new fashion. PMID- 15356711 TI - A manganese superoxide dismutase mimic based on cis,cis-1,3,5 triaminocyclohexane. AB - A novel manganese(iii) complex with a water soluble cis,cis-1,3,5 triaminocyclohexane-based ligand was synthesised and shown to exhibit superoxide dismutase activity. PMID- 15356712 TI - Synthesis and molecular structures of nitrosoarene metalloporphyrin complexes of ruthenium. AB - Several new ruthenium porphyrins containing nitrosoarene ligands have been synthesized and characterized by IR and (1)H NMR spectroscopy, and by single crystal X-ray crystallography. Bis-nitrosoarene complexes of the form (por)Ru(ArNO)(2)(Ar = aryl group; por = TPP, TTP; TPP = tetraphenylporphyrinato dianion, TTP = tetratolylporphyrinato dianion) were prepared in good yields from the reaction of the nitrosoarenes with (por)Ru(CO). The IR spectra of the complexes (as KBr pellets) display new bands in the 1346-1350 cm(-1) region due to nu(NO). Reactions of the (por)Ru(ArNO)(2) complexes with excess pyridine and 1 methylimidazole produce the mono-nitrosoarene complexes (por)Ru(ArNO)(py) and (por)Ru(ArNO)(1-MeIm), respectively. The IR spectra of these mono-nitrosoarene complexes reveal a lowering of nu(NO) by 14-44 cm(-1), a feature consistent with the replacement of one of the pi-acid ArNO ligands with the more basic pyridine and 1-MeIm ligands. The solid-state molecular structures of two members of each of the three classes of compounds, namely (por)Ru(ArNO)(2), (por)Ru(ArNO)(py) and (por)Ru(ArNO)(1-MeIm) were determined by single-crystal X-ray diffraction, and reveal the N-binding mode of the ArNO ligands. PMID- 15356713 TI - Gas phase ion chemistry of charged silver(I) adenine ions via multistage mass spectrometry experiments and DFT calculations. AB - Electrospray ionization (ESI) of solutions containing adenine and AgNO(3) yields polymeric [Ad(x)+ Ag(y)-zH]((y-z)+) species. Density functional theory (DFT) calculations have been used to examine potential structures for several of the smaller ions while multistage mass spectrometry experiments have been used to probe their unimolecular reactivity (via collision-induced dissociation (CID)) and bimolecular reactivity (via ion-molecule reactions with the neutral reagents acetonitrile, methanol, butylamine and pyridine). DFT calculations of neutral adenine tautomers and their silver ion adducts provide insights into the binding modes of adenine. We find that the most stable [Ad + Ag](+) ion does not correspond to the most stable neutral adenine tautomer, consistent with previous studies that have shown that transition metal ions can stabilize rare tautomeric forms of nucleobases. Both the charge and the stoichiometry of the [Ad(x)+ Ag(y) zH]((y-z)+) complexes play pivotal roles in directing the types of fragmentation and ion-molecule reactions observed. Thus, [Ad(2)+ Ag(2)](2+) is observed to dissociate to [Ad + Ag](+) and to react with butylamine via proton transfer, while [Ad(2)+ Ag(2)- H](+) fragments via loss of neutral adenine to form the [Ad + Ag(2)- H](+) ion and does not undergo proton transfer to butylamine. DFT calculations on several isomeric [Ad(2)+ Ag(2)](2+) ions suggest that planar centrosymmetric cations, in which two adjacent silver atoms are bridged by two N7H adenine tautomers via N(3),N(9)-bidentate interactions, are the most stable. The [Ad + Ag(2)-H](+) ion adds two neutral reagents in ion-molecule reactions, consistent with the presence of two vacant coordination sites. It undergoes a silver atom loss to form the [Ad + Ag - H](+) radical cation, which in turn fragments quite differently to the even electron [Ad + Ag](+) ion. Several other pairs of radical cation/even electron adenine-silver complexes were also found to undergo different fragmentation reactions. PMID- 15356714 TI - Generation of AuPd22/Au2Pd21 analogues of the high-nuclearity Pd23(CO)20(PEt3)10 cluster containing 19-atom centered hexacapped-cuboctahedral (nu 2-octahedral) metal fragment: structural-to-synthesis approach concerning formation of Au2Pd21(CO)20(PEt3)10. AB - Reactions of Pd(PEt(3))(2)Cl(2) and Au(PPh(3))Cl in DMF with NaOH under CO atmosphere gave rise to the unique capped three-shell homopalladium Pd(145)(CO)(x)(PEt(3))(30)(x approximately 60) and two neutral Au-Pd clusters: Au(2)Pd(21)(CO)(20)(PEt(3))(10) (1) and Au(2)Pd(41)(CO)(27)(PEt(3))(15)(following article). Similar reactions with Pd(PMe(3))(2)Cl(2) being used in place of Pd(PEt(3))(2)Cl(2) afforded Au(2)Pd(21)(CO)(20)(PMe(3))(10) (2), the trimethylphosphine analogue of, and the electronically equivalent [AuPd(22)(CO)(20)(PPh(3))(4)(PMe(3))(6)](-) monoanion (3) as the [PPh(4)](+) salt. Each of these three air-sensitive 23-atom heterometallic Au-Pd clusters was obtained in low yields (7-25%); however, their geometrical similarities with the known cuboctahedral-based homopalladium Pd(23)(CO)(20)(PEt(3))(10) (4), recently obtained in good yields from Pd(10)(CO)(12)(PEt(3))(6), suggested an alternative preparative route for obtaining. This "structure-to-synthesis" approach afforded 1 in 60-70% yields from reactions of Pd(10)(CO)(12)(PEt(3))(6) and Au(PPh(3))Cl in DMF with NaOH under N(2) atmosphere. Both the compositions and atomic arrangements for 1, 2 and 3 were unambiguously established from low-temperature single-crystal CCD X-ray crystallographic determinations in accordance with their nearly identical IR carbonyl frequencies. Cluster 1 was also characterized by (31)P[(1)H] NMR, cyclic voltammetry (CV) and elemental analysis. The virtually identical Au(2)Pd(21) core-architectures of 1 and 2 closely resemble that of 4, which consists of a centered hexa(square capped)-cuboctahedral Pd(19) fragment of pseudo-O(h) symmetry that alternatively may be viewed as a centered Pd(19)nu(2) octahedron (where nu(n) designates (n + 1) equally spaced atoms along each edge). [AuPd(22)(CO)(20)(PPh(3))(4)(PMe(3))(6)](-) (3) in the crystalline state ([PPh(4)](+) salt) consists of two crystallographically independent monoanions 3A and 3B; a superposition analysis ascertained that their geometries are essentially equivalent. A CV indicates that reversibly undergoes two one-electron reductions and two one-electron oxidations; these reversible redox processes form the basis for an integrated structural/electronic picture that is compatible with the existence of the electronically-equivalent 1-3 along with the electronically nonequivalent 4 (with two fewer CVEs) and other closely related species. PMID- 15356715 TI - Nanosized Au2Pd41(CO)27(PEt3)15 containing two geometrically unprecedented 13 coordinated Au-centered (mu 13-Au)Pd13 polyhedra connected by triangular face sharing and three interpenetrating 12-coordinated Pd-centered (mu 12-Pd)Au2Pd10 icosahedra: geometrical change in centered polyhedra induced by Au/Pd electronegativity-mismatch. AB - The synthesis, isolation, and stereochemical characterization of Au(2)Pd(41)(CO)(27)(PEt(3))(15)(1) are described. This nanosized Au(2)Pd(41) cluster (maximum metal-core diameter, 1.04 nm) was originally obtained with Au(2)Pd(21)(CO)(20)(PEt(3))(10) as low-yield by-products together with Pd(145)(CO)(x)(PEt(3))(30)(x approximately 60) from the reaction of Pd(PEt(3))(2)Cl(2) and Au(PPh(3))Cl in DMF with NaOH under CO atmosphere. The subsequent preparation of Au(2)Pd(21)(CO)(20)(PEt(3))(10) in greatly improved yields (preceding article) thereby provided the starting material that led to the isolation of 1 in reasonable yields (54%) from an overnight refluxing of the preformed Au(2)Pd(21) cluster in THF under N(2). Both the composition (subsequently ascertained from elemental analysis) and molecular geometry of 1 were unequivocally established from a low-temperature CCD X-ray diffraction study, which revealed a cubic unit cell of P2(1)3 symmetry with four molecules of 1 and four co-crystallized triphenylphosphine oxide molecules each lying on a crystallographic three-fold axis. The entire Au(2)Pd(41) core of pseudo-C(3h) symmetry may be viewed as a central Au(2)Pd(29) fragment of pseudo-D(3h) symmetry composed of two heretofore geometrically unknown 13-coordinated Au-centered (mu(13)-Au)Pd(13) polyhedra that share a common internal Pd(i)(3) triangular face perpendicular to the C(3) principal axis and of three three-fold-related interpenetrating 12-coordinated Pd-centered (mu(12)-Pd)Au(2)Pd(10) icosahedra. A comparative analysis of this central Au(2)Pd(29) fragment in with an internal Au(i)(2)Pd(i)(3) trigonal bipyramid vs. the corresponding central Pd(29) fragment in the known homopalladium Pd(35)(CO)(23)(PMe(3))(15) (2) with an internal Pd(i)(5) trigonal bipyramid resulting from five interpenetrating 12-coordinated Pd-centered [(mu(12)-Pd)Pd(12)] icosahedra is particularly illuminating; it provides a striking illustration of the remarkable observed difference between Pd vs. Au-centered polyhedra which is attributed to a large electronegativity mismatch in radial bonding interactions that occurs upon replacement of the Pd centered atom with a highly electronegative Au-centered atom. The entire Au(2)Pd(41) core-geometry is obtained by additional face-condensations of 12 tetracapping Pd(cap) atoms. This cluster is stabilized by 15 PEt(3) ligands and 27 doubly- and triply-bridging CO ligands. A close geometrical resemblance between the three three-fold-related Au(2)Pd(14) moities within the Au(2)Pd(41) core in 1 and the entire Au(2)Pd(14) core in the known [Au(2)Pd(14)(CO)(9)(PMe(3))(11)](2+) dication (3) is observed; resulting stereochemical implications are given. PMID- 15356716 TI - Structural and magnetic properties of vanadyl dichloride solvates: from molecular units to extended hydrogen-bonded solids. AB - The preparation, structural characterization and magnetic properties of three solvent adducts of VOCl(2), trans-VOCl(2)(THF)(2)(H(2)O) (1), trans VOCl(2)(H(2)O)(2).2Et(2)O (2) and cis-VOCl(2)(MeOH)(3) (3) are described. In these solids, hydrogen bonding among the inorganic complexes is the critical determinant of the formation of extended magnetic networks. Compound forms one dimensional double chains where alternating monomers from the two branches of the chain are hydrogen bonded via the V-Cl ... H-O-V network (with an axial water molecule and equatorial chloride ions). Magnetic studies indicate no interaction among the vanadyl centers. The paramagnetism of 1 is consistent with the extension of the network from the hydrogen donor site of the axial water, which is orthogonal to the d(xy) magnetic orbital. Compound 2 forms one-dimensional chains with water molecules of adjacent monomers held together by hydrogen bonds to ether molecules (V-O-H ... O(ether) ... H -O-V). The chain network radiates only through the equatorial plane of the complex where the water molecules are located. The presence of the intervening solvent molecule between hydrogen bonds of the primary coordination sphere magnetically insulates metal centers and compound is also a simple paramagnet. Removal of the solvent turns on the magnetic interaction and neighboring spin centers couple antiferromagnetically. Compound 3 forms a layered structure via V-Cl ... H-O-V hydrogen bonding, where all the hydrogen donor sites participate in the formation of the network. The vanadyl spin centers, at distances of 5.5 and 6.5 A from each other, couple antiferromagnetically (J/k=-0.7 K). Thus, magnetic coupling among metal centers is achieved when the hydrogen bond network directly radiates from the coordination plane containing the magnetic orbital. These results further support the utility of hydrogen bond as a viable design element in the construction of low dimensional, magnetic solids. PMID- 15356717 TI - Steric complementarity in carboxylate anion recognition. AB - A 2,6 pyridino-linked aminocryptand acts as a good host for carboxylate anions, oxalate, malonate, acetate and lactate at low protonation levels as shown by aqueous complexation constants. With oxalate, at the hexaprotonated level of the host, the crystal structure shows formation of a dimeric unit linked by an oxalate/semioxalate short (2.474 A) H-bond. The protonated cryptand uses a cleft binding mode in preference to the cavity-binding mode used by other aminocryptand hosts. PMID- 15356718 TI - A crystallographic, EPR and theoretical study of the Jahn-Teller distortion in [CuTp2] (Tp- = tris[pyrazol-1-yl]hydridoborate). AB - Crystals of the title compound (1) contain two independent, centrosymmetric half molecules per asymmetric unit. While both of these show Jahn-Teller elongated six coordinate geometries, the lengths of the elongated Cu-N bonds in the two molecules differ by 0.117(2) A at 30 K. The structure of one of these molecules (molecule A) does not vary with temperature below 350 K. The other molecule (molecule B) shows Cu-N bond lengths that are temperature-dependent between 225 and 375 K, but do not vary further at lower temperature. This indicates a fluxional axis of Jahn-Teller elongation in this molecule at these higher temperatures. Consideration of the thermal parameters in these structures implies that the fluxionality in molecule B is frozen out near 150 K. This conclusion is supported by a Q-band powder EPR study. The d-d transition energies of molecules A and B have been calculated by several density function (DF) methods, including a time-dependent DF calculation. The crystallographic data have been reproduced using the vibronic coupling model of Burgi and Hitchman. This has shown that the different fluxionality regimes for molecules A and B are not a consequence of their different static molecular structures, but rather reflect their different local environments in the crystal. PMID- 15356719 TI - Solvent extraction and lanthanide complexation studies with new terdentate ligands containing two 1,3,5-triazine moieties. AB - The extracting agent 2,6-bis(4,6-di-pivaloylamino-1,3,5-triazin-2-yl)-pyridine (L(5)) in n-octanol was found, in synergy with 2-bromodecanoic acid, to give D(Am)/D(Eu) separation factors (SFs) between 2.4 and 3.7 when used to extract the metal ions from 0.02-0.12 M HNO(3). Slightly higher SFs (4-6) were obtained in the absence of the synergist when the ligand was used to extract Am(III) and Eu(III) from 0.98 M HNO(3). In order to investigate the possible nature of the extracted species crystal structures of L(5) and the complex formed between Yb(III) with 2,6-bis(4,6-di-amino-1,3,5-triazin-2-yl)-pyridine (L(4)) were also determined. The structure of L(5) shows 3 methanol solvent molecules all of which form 2 or 3 hydrogen bonds with triazine nitrogen atoms, amide nitrogen or oxygen atoms, or pyridine nitrogen atoms. However, L(5) is relatively unstable in metal complexation reactions and loses amide groups to form the parent tetramine L(4). The crystal structure of Yb(L(4))(NO(3))(3) shows ytterbium in a 9-coordinate environment being bonded to three donor atoms of the ligand and three bidentate nitrate ions. The solvent extraction properties of L(4) and L(5) are far inferior to those found for the 2,6-bis-(1,2,4-triazin-3-yl)-pyridines (L(1)) which have SF values of ca. 140 and theoretical calculations have been made to compare the electronic properties of the ligands. The electronic charge distribution in L(4) and L(5) is similar to that found in other terdentate ligands such as terpyridine which have equally poor extraction properties and suggests that the unique properties of L(1) evolve from the presence of two adjacent nitrogen atoms in the triazine rings. PMID- 15356720 TI - Different supramolecular hydrogen bond structures and significant changes in magnetic properties in dinuclear mu 2-1,1-N3 copper(II) complexes with very similar tridentate Schiff base blocking ligands. AB - Three new basal-apical, mu(2)-1,1-azide bridged complexes, [CuL(1)(N(3))](2) (1), [CuL(2)(N(3))](2) (2) and [CuL(3)(N(3))](2) (3) with very similar tridentate Schiff base blocking ligands [L(1) = N-(3-aminopropyl)salicylaldimine, L(2) = 7 amino-4-methyl-5-azahept-3-en-2-one and L(3) = 8-amino-4-methyl-5-azaoct-3-en-2 one) have been synthesised and their molecular structures determined by X-ray crystallography. In complex 1, there is no inter-dimer H-bonding. However, complexes 2 and 3 form two different supramolecular structures in which the dinuclear entities are linked by strong H-bonds giving one-dimensional systems. Variable-temperature (300-2 K) magnetic susceptibility measurements and magnetization measurements at 2 K reveal that complexes and have antiferromagnetic coupling while has ferromagnetic coupling which is also confirmed by EPR spectra at 4-300 K. Magnetostructural correlations have been made taking into consideration both the azido bridging ligands and the existence of intermolecular hydrogen bonds in complexes 2 and 3. PMID- 15356721 TI - Synthesis, structural analysis, and magnetic behaviour of three fumarate bridged coordination polymers: five-fold interpenetrated diamond-like net of NiII, sheets of NiII and CoII. AB - The hydrothermal reactions of Ni(NO(3))(2).6H(2)O, disodium fumarate (fum) and 1,2-bis(4-pyridyl)ethane (bpe)/1,3-bis(4-pyridyl)propane (bpp) in aqueous methanol medium yield one 3-D and one 2-D metal-organic hybrid material, [Ni(fum)(bpe)] (1) and [Ni(fum)(bpp)(H(2)O)] (2), respectively. Complex possesses a novel unprecedented structure, the first example of an "unusual mode" of a five fold distorted interpenetrated network with metal-ligand linkages where the four six-membered windows in each adamantane-type cage are different. The structural characterization of complex 2 evidences a buckled sheet where nickel ions are in a distorted octahedral geometry, with two carboxylic groups, one acting as a bis chelate, the other as a bis-monodentate ligand. The metal ion completes the coordination sphere through one water molecule and two bpp nitrogens in cis position. Variable-temperature magnetic measurements of complexes 1 and 2 reveal the existence of very weak antiferromagnetic intramolecular interactions and/or the presence of single-ion zero field splitting (D) of isolated Ni(II) ions in both the compounds. Experimentally, both the J parameters are close, comparable and very small. Considering zero-field splitting of Ni(II), the calculated D values are in agreement with values reported in the literature for Ni(II) ions. Complex 3, [[Co(phen)]](2)(fum)(2)](phen = 1,10-phenanthroline) is obtained by diffusing methanolic solution of 1,10-phenanthroline on an aqueous layer of disodium fumarate and Co(NO(3))(2).6H(2)O. It consists of dimeric Co(II)(phen) units, doubly bridged by carboxylate groups in a distorted syn-syn fashion. These fumarate anions act as bis-chelates to form corrugated sheets. The 2D layer has a (4,4) topology, with the nodes represented by the centres of the dimers. The magnetic data were fitted ignoring the very weak coupling through the fumarate pathway and using a dimer model. PMID- 15356722 TI - Sulfite induced autoxidation of NiII and CoII tetraglycine complexes. Spectrophotometric and rotating ring-disc voltammetric studies. AB - The oxidation of Ni(II) and Co(II) tetraglycine complexes in borate buffer aqueous solution, by dissolved oxygen, is strongly accelerated by sulfite. The formation of Ni(III) and Co(III) complexes with maximum absorbances at 327 and 265 nm, respectively, was followed by spectrophotometric measurements. Ni(III) formation was also characterized by voltammetry at low temperatures, whose anodic and cathodic components were observed in the recorded voltammograms. Spectra and rotating ring-disc voltammograms, recorded at various rotation speed values, showed that the Ni(III) species decomposes. The electrochemical process related to the couple Co(II)/Co(III), in a medium containing tetraglycine, was not reversible. In both Ni(II) and Co(II) complexes the metal ion oxidation in the presence of oxygen and sulfite involves the reduction of some initial Ni(III) or Co(III) by sulfite to produce the SO(3).- radical, which rapidly reacts with dissolved oxygen to produce SO(5).-, which then oxidizes Ni(II) or Co(II). PMID- 15356723 TI - Synthesis, X-ray structure, polarized optical spectra and DFT theoretical calculations of two new organic-inorganic hybrid fluoromanganates(III): (bpaH2)[MnF4(H2O)2]2 and (bpeH2)[MnF4(H2O)2]2. AB - Two new fluoromanganates(III) of 1,2-bis(4-pyridyl)ethane (bpa) and trans-1,2 bis(4-pyridyl)ethylene (bpe), LH(2)[MnF(4)(H(2)O)(2)](2) (L = bpa or bpe), have been prepared and their structure have been solved by single-crystal X-ray diffraction. The [MnF(4)(H(2)O)(2)](-) anion displays an octahedral geometry with a strong Jahn-Teller tetragonal distortion along the H(2)O-Mn-OH(2) axis. The equatorial metal-ligand distances (Mn-F 1.827(1)-1.859(2) A) are shorter than the axial ones (Mn-O 2.203(2)-2.234(2) A). Three polarized absorption bands at 22,500, 18,300 and 14,500 cm(-1) are observed in the optical spectra of (bpaH(2))[MnF(4)(H(2)O)(2)](2). Finally, we present theoretical calculations on the equilibrium bond distances as well as the crystal-field electron structure using density functional methods. The calculated Mn-F bond distances (1.85 A) are in agreement with the experimental data but the obtained Mn-O distances (2.53 2.56 A) are higher than the experimental one as usually found in similar Jahn Teller distorted systems. The calculated d-d transition energies are compared with experimental energies derived from the optical spectra. The variation of the HOMO energy and transition energies against the Mn-O distance is also shown. PMID- 15356724 TI - Substitution reactions of [Pt(terpy)X]2+ with some biologically relevant ligands. Synthesis and crystal structure of [Pt(terpy)(cyst-S)](ClO4)2.0.5H2O and [Pt(terpy)(guo-N7)](ClO4)2.0.5guo.1.5H2O. AB - Substitution reactions of the complexes [Pt(terpy)(H(2)O)](2+), [Pt(terpy)(cyst S)](2+) and [Pt(terpy)(guo-N(7))](2+), where terpy = 2,2':6',2"-terpyridine, cyst = L-cysteine and guo = guanosine, with some biologically relevant ligands such as inosine (INO), inosine-5'-monophosphate (5'-IMP), guanosine-5'-monophosphate (5' GMP), l-cysteine, glutathione, thiourea, thiosulfate and diethyldithiocarbamate (DEDTC), were studied in aqueous 0.10 M NaClO(4) at pH 2.5 and 6.0 using variable temperature and -pressure stopped-flow spectrophotometry. The reactions of [Pt(terpy)(H(2)O)](2+) with INO, 5'-IMP and 5'-GMP showed that these ligands are very good nucleophiles. The second order rate constants varied between 4 x 10(2) and 6 x 10(2) M(-1) s(-1) at 25 degree C. The [Pt(terpy)(cyst-S)](2+) complex is unreactive towards nitrogen donor nucleophiles, and cysteine cannot be replaced by N(7) from INO, 5'-IMP and 5'-GMP. However, sulfur donor nucleophiles such as thiourea, thiosulfate and diethyldithiocarbamate could displace the Pt-cysteine bond. Diethyldithiocarbamate is the best nucleophile and the order of reactivity is: thiourea < thiosulfate < DEDTC with rate constants of 0.936 +/- 0.002, 5.99 +/- 0.02 and 8.88 +/- 0.07 M(-1) s(-1) at 25 degree C, respectively. The reactions of [Pt(terpy)(guo-N(7))](2+) with sulfur donor ligands showed that these nucleophiles could substitute guanosine from the Pt(ii) complex, of which diethyldithiocarbamate and thiosulfate are the strongest nucleophiles. The tripeptide glutathione is also a very efficient nucleophile. Activation parameters (Delta H(++), Delta S(++) and Delta V(++)) were determined for all reactions. The crystal structures of [Pt(terpy)(cyst-S)](ClO(4))(2).0.5H(2)O and [Pt(terpy)(guo-N(7))](ClO(4))(2).0.5guo.1.5H(2)O were determined by X-ray diffraction. Crystals of [Pt(terpy)(cyst-S)](ClO(4))(2).0.5H(2)O are orthorhombic with the space group P2(1)2(1)2(1), whereas [Pt(terpy)(guo N(7))](ClO(4))(2).0.5guo.1.5H(2)O crystallizes in the orthorhombic space group P2(1)2(1)2. A typical feature of terpyridine complexes can be found in both molecular structures: the Pt-N (central) bond distance, 1.982(7) and 1.92(2) A, respectively, is shorter than the other two Pt-N distances, being 2.043(7) and 2.034(7) A in [Pt(terpy)(cyst-S)](ClO(4))(2).0.5H(2)O and 2.03(2) and 2.04(2) A in [Pt(terpy)(guo-N(7))](ClO(4))(2).0.5guo.1.5H(2)O, respectively. In both crystal structures two symmetrically independent cations representing different conformers are present in the asymmetric unit. The results are analysed in reference to the antitumour activity of Pt(II) complexes, and the importance of the rescue agents are discussed. PMID- 15356725 TI - Electronically and sterically tuned trans labilization controls the substitution behaviour of cobaloximes. AB - The kinetics of axial water substitution by cysteine in six different cobaloximes, viz.trans-RCo(Hdmg)(2)H(2)O, where Hdmg = dimethylglyoximate, R = cyclo-C(5)H(9) (c-P), CH(3)CH(2) (Et), CH(3) (Me), C(6)H(5)CH(2) (Bz), C(6)H(5) (Ph) and CF(3)CH(2), were studied as a function of cysteine concentration, temperature and pressure. It was found that cysteine substitutes the coordinated H(2)O molecule trans to the alkyl group with second order rate constants that follow the order of reactivity: c-P > Et > Bz > Me > Ph > CF(3)CH(2). Rate and activation parameters (Deltan H(++), Delta S(++) and Delta V(++)) enable the formulation of a reaction mechanism that can account for the substitution behaviour of the investigated alkylcobaloximes. In particular, a gradual mechanistic changeover from I(d) to I is observed along the series of R groups from c-P to CF(3)CH(2). PMID- 15356726 TI - Mechanistic information on the copper-catalysed autoxidation of mercaptosuccinic acid in aqueous solution. AB - Copper(II) ions react rapidly with sulfur from thiol groups, forming two distinct, intensely absorbing, short-lived intermediates, which decompose in a subsequent redox reaction to produce reduced copper and disulfides. In this study we report the results of a mechanistic study on the reaction between mercaptosuccinic acid, HO(2)CCH(2)CH(SH)CO(2)H, and Cu(2+)(aq) and [Cu(tren)H(2)O](2+), tren = tris(2-aminoethyl)amine. Spectroscopic and kinetic data indicate that in the presence of an excess of thiol, at least two distinct complexes are formed, with very different decomposition rate constants and an absorption maximum at 346 nm. Upon addition of thiol to [Cu(tren)H(2)O](2+)(1:1), a transient with a maximum at 380 nm appears, whereas in an excess of thiol this complex decomposes and again the 346 nm band is observed. The use of [Cu(tren)H(2)O](2+) enables to study the reaction of thiol with copper also in alkaline solution, where the rate of the overall process is slowed down greatly. The reactions were studied in detail, including the effect of dioxygen, and a possible reaction mechanism for the catalysed autoxidation process is proposed and discussed in reference to available literature data. PMID- 15356727 TI - Role of chelate substituents and cis sigma-effect on the rate of ligand substitution at Pt(N-N-N) and Pt(N-N-C) centres. AB - Four complexes of the type [Pt(N-N-X)Cl](X = N or C) were tailor synthesized for mechanistic studies in methanol. The terdentate ligands included terpy, 4'-Ph terpy, 4'-(2"-CF(3)-Ph)-terpy, and 4'-(2"'-CF(3)-Ph)-6-Ph-2,2'-bipy. The rate of substitution of the chloro ligand by thiourea, N,N'-dimethylthiourea, and N,N,N',N'-tetramethylthiourea was studied as a function of nucleophile concentration, temperature and pressure by using a stopped-flow technique. The observed pseudo-first-order rate constants for the substitution reactions obeyed the simple rate law k(obs) = k(2)[Nu]. Second-order kinetics and negative activation entropies and volumes support an associative substitution mechanism. At 298 K, the values of the second-order rate constant show a slight dependence on the nature of the moiety attached to the terpy ligand. Changing from a nitrogen sigma-donor to a carbon sigma-donor in the cis position, results in a deceleration of the substitution rate. The results suggest that the Pt-C bond in the cis position activates the metal centre in a different way than in the trans position. PMID- 15356728 TI - Two-electron reduction of alkyl(sulfanyl)porphyrazines: a route to free-base and peripherally metallated asymmetric porphyrazines. AB - Treatment of octakis(octylthio)porphyrazine (H(2)OOSPz) with cis bis(benzonitrile)dichloroplatinum(II) in a 1,2,4-trichlorobenzene (TCB)-n-BuOH mixture at 190 degree C affords the asymmetric porphyrazine 2H heptakis(octylthio)porphyrazine, H(2)HOSPz, in ca. 40% yield. In the key synthetic step the two-electron reduced diprotonated porphyrazine, [H(2)OOPz(4 )(H(+))(2)](hemiporphyrazinogen), forms in nearly quantitative yield. This species undergoes nucleophilic substitution of an octylsulfide group by H(-), affording the asymmetric porphyrazine, this process being favoured by the high temperature induced charge and structural intramolecular rearrangements. The resulting molecule shows mesomorphic behaviour. Interestingly, in the presence of sodium acetate, or in neat TCB, the asymmetric porphyrazine does not form at all. Under these conditions, a PtCl(2) molecular fragment coordinates both to a porphyrazine aza bridge and to a thioether moiety leading to the formation of the [H(2)OOSPz]PtCl(2) complex. The S,N-coordination of the PtCl(2) unit was proved by (1)H, (195)Pt[(1)H], and (13)C NMR spectroscopy, and supported by DFT (B3LYP) calculations. The complex conjugates high asymmetry with strong solvatochromism and, therefore, it is potentially interesting for nonlinear optics. PMID- 15356729 TI - Zwitterionic macrocyclic metal sulfate extractants containing 3 dialkylaminomethylsalicylaldimine units. AB - 10,25-Di-tert-alkyl-14,21-dimethyl-3,6,14,21 tetraazatricyclo[21.3.1.18,12]octacosa-1(27),2,6,8,10,12(28),23,25-octaene-27,28 diol macrocycles form Ni(II) and Cu(II) complexes in which the metal cation and the sulfate anion are bound in separated sites in a zwitterionic form of the ligand. The nonyl-substituted macrocycle shows a higher affinity for SO(4)(2-) and a lower binding strength for Cu(2+) than open chain analogues, the pH dependences for which fall in ranges which allow loading of CuSO(4) at pH approximately 4 and easy stripping to recycle the ligand. X-Ray structure determinations of the Cu(II) and Ni(II) sulfate complexes of the tert-butyl substituted ligand suggest that the de-tuning of M(2+)-binding results from a distortion from planarity of the "salen" N(2)O(2)(2-) donor set imposed by the incorporation of the hexamethylene strap in the ligand and reveal that the sulfate is bound as a hydrate in a 2:2:2:2, ligand-M(2+)-SO(4)(2-)-H(2)O, assembly. PMID- 15356730 TI - Metallation effects on the thermal interconversion of atropisomers of di(orthomethylarene)-substituted porphyrins. AB - A new series of meso-substituted diaryl free-base and metalloporphyrins have been prepared. Each arene has been substituted with both a methyl group in the ortho position and a formyl group in the meta position. Rotation of the arene units is prevented at room temperature due to the steric restrictions imposed by the flanking methyl groups at the porphyrin beta-pyrrolic positions on the methyl groups at the ortho position on the meso-substituted arene unit. This allowed the alpha alpha and alpha beta atropisomers of this porphyrin to be separated and characterised. X-Ray crystallographic determination of the structure of the free base porphyrin revealed a very flat porphyrin core. Metallation resulted in the isolation and characterisation of the nickel, zinc and copper derivatives. The assignments of the alpha alpha and alpha beta isomers are confirmed by X-ray crystallographic determination of the structures of the Cu(II) analogues. The copper alpha alpha structure exhibits a very twisted porphyrin core, the copper alpha beta structure is also distorted, but to a lesser degree. The activation energy for rotation has been calculated for each of the 2H, Ni and Zn derivatives. The energy required to rotate the arene ring increases in the order Ni < Zn approximately 2H. No significant difference in the free energy of rotation was observed between experiments carried out with the alpha alpha and small alpha beta isomers. PMID- 15356731 TI - A paradigm shift in the construction of heterobimetallic complexes: synthesis of group 2 & 4 metal-calix[6]arene complexes. AB - Deprotonation of calix[6]arenes with barium in methanol followed by the addition of [Ti(OPr(i))(4)] or [Zr(OBu(n))(4)] is effective in the formation of novel dimeric 2:1 barium-titanium(IV)/zirconium(IV) calix[6]arene complexes. In these complexes a central Ti(IV)/Zr(IV) coordinated in the exo-position connects the two calix[6]arenes in the 1,3-alternate conformation, each with an endo-barium sharing common phenolate groups with the titanium/zirconium centre and participating in cation-pi interactions. A homometallic barium calix[6]arene dimer was also prepared wherein the calix[6]arenes are in the 1,3-alternate conformation with each coordinating one endo- and one exo-barium centre. The exo barium cations connect the two calix[6]arenes through bridging methanol ligands. In this and the heterometallic complexes, cation-pi complexation of the Ba(2+) ion within the 1,3 alternate conformation of calix[6]arene facilitates the formation of the dimeric complexes in methanol. In contrast, the smaller Sr(2+) ion did not form similar complexes in methanol, and the formation of an analogous 2:1 strontium-titanium calixarene complex required the use of the more sterically demanding donor alcohol, isopropanol, the resulting complex being devoid of cation-pi interaction. The results show (i) that a subtle interplay of solvation strength, coordination array type and cavity/cation size influences the accessibility of heterobimetallic complexes based on calix[6]arenes, and (ii) a synergistic endo-exo binding behaviour. PMID- 15356732 TI - Electronic properties of Ru(II) complexes bound to a bisphenolate bridge with low lying pi* orbitals. AB - The synthesis and a detailed investigation into the electronic properties of mononuclear and dinuclear Ru(II) complexes of the ligand bis(2-hydroxyphenyl)-2,5 dihydropyrazine (H(2)BHD) is described. In these complexes the Ru(II) moieties are bound through O,N coordination to an anionic phenolate and the pyrazine bridge. Relatively few reports are available on the dinuclear complexes bridged across a phenolate and this study provides an opportunity to examine the impact of reduced oxygen donor ligands on metal-metal communication. The results presented here reveal some very unusual behavior whereby the apparent location of the LUMO changes between the mononuclear and dinuclear complexes. The lowest energy optical transition appears to involve the peripheral bipyridine ligand as acceptor in the mononuclear complex, whereas this ligand is not involved in the lowest energy optical transition in the dinuclear complex. The origin of this difference is not clear, however, significant changes in the electronic properties of the mononuclear complex are observed on coordination of the second metal, reflected in significant alterations in the electrochemistry of the bridge and metals as well as changes in the optical spectroscopy. The BHD(2-) bridge is shown to support weakly coupled class II behavior according to the Robin and Day classification, reflected in a K(c) of 335. PMID- 15356733 TI - Complex formation reaction of the iron(III) hydroxo dimer with periodate ion. AB - The kinetics and mechanism of the ligand substitution reaction between Fe(2)(OH)(2)(4+) and periodate ion has been studied. This process is unique among the reactions of the iron(iii) hydroxo dimer because the initial rate is second order with respect to Fe(2)(OH)(2)(4+). The formation of a bi- and a tetranuclear complex, Fe(2)(OH)(2)(H(4)IO(6))(3+) and Fe(4)(OH)(4)(H(4)IO(6))(7+), is proposed. Comprehensive fitting of the kinetic data was used to show that the proposed model, which is very similar to earlier models used with other inorganic oxoanions, gives a reasonable interpretation of all observations. It is shown that the lifetime of Fe(2)(OH)(2)(H(4)IO(6))(3+) is relatively long and it can open a pathway to form oligomeric and less soluble products at higher initial concentrations. The speciation of aqueous periodate ion solution was also studied and it is proposed that the tetrahedral form, IO(4)(-), is less dominant over the octahedral form, H(4)IO(6)(-), than previously thought. PMID- 15356734 TI - The profound influence of a single metal-carbon bond on the reactivity of bio relevant cobalt(III) complexes. AB - This Perspective reports the development of mechanistic insight over the past 6 years, on the substitution behaviour of cobalamins that contain a single Co-C bond. The effect of the alkyl group, located in the trans position, on the thermodynamic, kinetic and ground state trans effect, was studied in detail. The substitution reactions of different alkylcobalamins with CN- were investigated, the apparent mechanistic discrepancy reported for the co-enzyme B12 was resolved and a logical explanation could be offered. In addition, a complete picture of the effect of pressure on the UV-Vis spectra of different base-on and base-off cobalamins is presented, which clearly shows the role of the alkyl group in controlling the equilibrium between five- and six-coordinate species, and the possible participation of such species in the studied ligand substitution reactions. The kinetics of the base-on/base-off equilibration was studied for the first time using a pH-jump technique. All in all the novel mechanistic information adds to the understanding of the profound effect that a single metal carbon bond can have on the reactivity of such Co(III) complexes. PMID- 15356735 TI - Unusual photophysical switching in a Ru(II) diimine DNA probe caused by amide functionalisation. AB - Whereas the complex [Ru(phen)2Medpq]2+(phen=1,10-phenanthroline; Medpq=2 methyldipyrido[3,2-f:2',3'-h]-quinoxaline) is luminescent in both aqueous and nonaqueous solutions, [Ru(phen)2dpqa]2+(2-pentylamidodipyrido[3,2-f:2',3'-h] quinoxaline) does not emit in water but does so strongly in organic solvents or when bound to DNA--suggesting its use as a photochemical and photophysical probe for nucleic acids. PMID- 15356736 TI - Identification of a novel high affinity copper binding site in the APP(145-155) fragment of amyloid precursor protein. AB - The copper(II) binding features of the APP(145-155) and APP(145-157) fragments of the amyloid precursor protein, Ac-Glu-Thr-His-Leu-His-Trp-His-Thr-Val-Ala-Lys-NH2 and Ac-Glu-Thr-His-Leu-His-Trp-His-Thr-Val-Ala-Lys-Glu-Thr-NH2 were studied by NMR spectroscopy and NMR findings were supported by UV-vis, CD and EPR spectra. Potentiometric measurements were performed only for the more soluble Ac-Glu-Thr His-Leu-His-Trp-His-Thr-Val-Ala-Lys-Glu-Thr-NH2 peptide fragment. The following was shown: (i) the imidazole rings of all the three His residues are involved in metal coordination; (ii) metal binding induces ionisation of Leu-148 and His-149 amide nitrogens that complete the donor set to copper(II) in the species dominant at neutral pH; (iii) the unusual coordination scheme of the His-Xxx-His-Xxx-His consensus sequence justifies the high specificity for Cu(II) when compared to SOD like or albumin-like peptides or even in amyloid Abeta fragments. The present findings may represent the key for interpreting the observed requirement of His residues conservation for the redox cycling between Cu(II) and Cu(I) by soluble APP. PMID- 15356737 TI - Charge density analysis of two polymorphs of antimony(III) oxide. AB - High-resolution X-ray diffraction data have been collected on the cubic polymorph of antimony(III) oxide (senarmontite) to determine the charge distribution in the crystal. The results are in quantitative agreement with crystal Hartree-Fock calculations for this polymorph, and have been compared with theoretical calculations on the orthorhombic polymorph (valentinite). Information about the nature of bonding and relative bond strengths in the two polymorphs has been extracted in a straightforward manner via topological analysis of the electron density. All the close contacts in both polymorphs are found to be similar in nature based on the value of the Laplacian, the magnitude of the electron density and the local energy density at the bond critical points, and these characterise the observed interactions as substantially polar covalent, similar to molecular calculation results on Si-O and Ge-O. Electrostatic potential isosurfaces reveal the octopolar nature of this function for senarmontite, and shed light on the observed packing arrangement of Sb4O6 molecules in the crystal. PMID- 15356738 TI - Photosensitized generation of singlet oxygen from ruthenium(II) and osmium(II) bipyridyl complexes. AB - Photophysical properties for a number ruthenium(II) and osmium(II) bipyridyl complexes are reported in dilute acetonitrile solution. The lifetimes of the excited metal to ligand charge transfer states (MLCT) of the osmium complexes are shorter than for the ruthenium complexes. Rate constants, kq, for quenching of the lowest excited metal to ligand charge transfer states by molecular oxygen are found to be in the range (1.1-7.7) x 10(9) dm3 mol(-1) s(-1). Efficiencies of singlet oxygen production, fDeltaT, following oxygen quenching of the lowest excited states of these ruthenium and osmium complexes are in the range of 0.10 0.72, lower values being associated with those compounds having lower oxidation potentials. The rate constants for quenching of the excited MLCT states, kq, are found to be generally higher for osmium complexes than for ruthenium complexes. Overall quenching rate constants, kq were found to give an inverse correlation with the energy of the excited state being quenched, and also to correlate with the oxidation potentials of the complexes. However, when the contribution of quenching due exclusively to energy transfer to produce singlet oxygen, kq1, is considered, its dependence on the energy of the excited states is more complex. Rate constants for quenching due to energy dissipation of the excited MLCT states without energy transfer, kq3, were found to show a clear correlation with the oxidation potential of the complexes. Factors affecting both the mechanism of oxygen quenching of the excited states and the efficiency of singlet oxygen generation following this quenching are discussed. These factors include the oxidation potential, the energy of the lowest excited state of the complexes and spin-orbit coupling constant of the central metal. PMID- 15356739 TI - Dicatechol-diimines: easily accessible ligands for the self-assembly of dinuclear triple-stranded helicates. AB - Dicatechol ligands 3b-g-H4 are simply prepared by imine formation of 2,3 dihydroxybenzaldehyde 2 with a series of different diamines 1b-g . An X-ray structural analysis was obtained for the butyl-bridged compound 3e-H4, showing an intramolecular proton transfer and the formation of a chinoidic "keto-amine" structure. The dicatechol derivatives 3b-g-H4 form dinuclear triple-stranded helicates M4[(3)3Ti2] with titanium(IV) ions in the presence of alkali-metal carbonate. For the phenyl- and the trans-1,4-cyclohexyl-bridged complexes, K4[(3b)3Ti2] and Na4[(3f)3Ti2], X-ray structures were obtained. PMID- 15356741 TI - Synthesis and properties of new ditertiary stibines based upon o-, m- or p-xylyl and m- or p-phenylene backbones and their complexes with tungsten, iron and nickel carbonyls. AB - High yield syntheses for 1,2-, 1,3-, and 1,4-xylyl distibines (1,2 C6H4(CH2SbMe2)2, 1,3-C6H4(CH2SbMe2)2, 1,4-C6H4(CH2SbMe2)2, respectively) from Me2SbCl (conveniently made in situ from Me2PhSb and HClgas) and the appropriate di-Grignard are reported. The 1,3- and 1,4-phenylene distibines, 1,3-C6H4(SbMe2)2 and 1,4-C6H4(SbMe2)2, were made similarly. The new ligands have been characterised by mass spectrometry, 1H and 13C[1H] NMR spectroscopy, and by the preparation of methiodide derivatives. The crystal structures of 1,4 C6H4(CH2SbMe2)2 and [1,3-C6H4(CH2SbMe3)2]I2 have been determined. The synthesis of 1,2-C6H4(CH2SbPh2)2 has been achieved similarly in modest yield and the distibine converted into the tetra-iodo-derivative 1,2-C6H4(CH2SbPh2I2)2. The coordination modes available to these ligands have been probed by the synthesis and characterisation of complexes with nickel, iron and tungsten carbonyls. The crystal structure of [[Fe(CO)4]2[micro-1,3-C6H4(CH2SbMe2)2]] has been determined. The spectroscopic properties of these carbonyl derivatives have been compared with those of complexes of other antimony ligands, and in some cases with diphosphine and diarsine complexes, to probe the electronic properties of the new ligands. PMID- 15356740 TI - Synthesis, characterization and molecular structures of six-coordinate manganese nitrosyl porphyrins. AB - Manganese(II) porphyrins are isoelectronic with iron(III) porphyrins, and previously reported work suggests that manganese nitrosyl porphyrins are good structural models for their kinetically unstable and biologically relevant ferric NO analogues. We have prepared a new set of six-coordinate manganese nitrosyl porphyrins of the general form (por)Mn(NO)(L)(por = TTP, T(p-OCH3)PP; L = piperidine, methanol, 1-methylimidazole) in moderate to high yields. The (por)Mn(NO)(pip) complexes were prepared from the reductive nitrosylation of the (por)MnCl compounds with NO in the presence of piperidine. The IR spectra of the (por)Mn(NO)(pip) compounds as KBr pellets show new strong bands at 1746 cm( 1)(for TTP) and 1748 cm(-1)(for (T(p-OCH3)PP) due to the NO ligands. Attempted crystallization of one of these compounds (por = TTP) from dichloromethane methanol resulted in the generation of the methanol complex (TTP)Mn(NO)(CH3OH). Reaction of the (por)Mn(NO)(pip) compounds with excess 1-methylimidazole gave the (por)Mn(NO)(1-MeIm) derivatives in good yields. The IR spectra of these compounds show nu(NO) bands that are approximately 12 cm(-1) lower than those of the (por)Mn(NO)(pip) precursors, indicative of greater Mn-->NO pi-backdonation in the 1-MeIm derivatives. X-Ray crystal structures of three of these compounds, namely (TTP)Mn(NO)(CH3OH), (TTP)Mn(NO)(1-MeIm) and (T(p-OCH3)PP)Mn(NO)(1-MeIm) were obtained, and reveal that the NO ligands in these complexes are linear. PMID- 15356742 TI - Antisymmetric exchange in two tricopper(II) complexes containing a [Cu3(micro3 OMe)]5+ core. AB - Reaction of CuX2(X-=Cl- or Br-) with 2 molar equivalents of 3[5]-(2,4,6 trimethylphenyl)pyrazole (HpzMes) in MeOH in the presence of NaOH yields [Cu3X(HpzMes)2(micro-pzMes)3(micro3-OMe)]X (X-=Cl- or Br-). Crystal structures of these compounds show almost identical triangles of Cu(II) ions, centred by a triply bridging methoxide ligand and with three edge-bridging pyrazolide groups. The mesityl substituents on the bridging pyrazolide ligands are arranged in HT, HH, TT fashion. chi(M)T for both compounds decreases steadily with decreasing temperature, reaching 0.40 cm(3) mol(-1) K at 70 K before decreasing further below 40 K. This low temperature behaviour could not be interpreted using conventional superexchange Hamiltonians, but was reproduced by an alternative model that incorporated an additional antisymmetric exchange term. This interpretation was confirmed by the Q-band EPR spectra of the two compounds. NMR experiments show that the structures of these compounds are not retained in solution, in contrast to other closely related tricopper compounds. These are the first examples of triangular Cu(II) compounds bearing a [Cu3micro3-OR)]5+(R is not equal to H) core motif, and the first triangular compounds showing antisymmetric exchange to have been analysed by both susceptibility and EPR measurements. PMID- 15356743 TI - A study of the thermal and light induced spin transition in [FeL2](BF4)2 and [FeL2](ClO4)2 L=2,6-di(3-methylpyrazol-1-yl)pyrazine. AB - The spin crossover compounds [FeL2](BF4)2, L=2,6-di(3-methylpyrazol-1-yl)pyrazine and [FeL2](ClO4)2 have very unusual two stage spin transitions which are initially steep and then become more gradual. A detailed variable temperature single crystal X-ray diffraction study has shown that the course of the spin transition is controlled by an order-disorder transition in the counter anions. The high and low spin states both crystallise in the tetragonal space group I4, the structures of the high and low spin states are presented at 290 and 30 K, respectively. The title compounds are shown to undergo LIESST (Light Induced Excited Spin State Trapping) under irradiation with either red or green laser light with wavelengths of 632.8 and 532.06 nm, respectively, at 30 K. The cell parameters for the tetragonal photo-induced metastable high spin state at this temperature are a= 9.169(6), c= 17.77(1) A for [FeL2](ClO4)2 with an increase in unit cell volume of 21 A3, and a= 9.11(1), c= 17.75(2) A and an increase in volume of 42.8 A3 for [FeL2](BF4)2. PMID- 15356744 TI - Structural and NMR investigations of the ternary adducts of twenty alpha-amino acids and selected dipeptides with a chiral, diaqua-ytterbium complex. AB - A detailed investigation of the nature of the binding of each of the 20 common alpha-amino acids and various selected dipeptides to a chiral, diaqua-ytterbium complex in aqueous solution has been carried out. Analysis of the dipolar 1H NMR paramagnetic shifts suggests that the alpha-amino acids form a common chelated structure within a nine-coordinate mono-capped square antiprismatic coordination environment, with the amine N axially disposed. Crystal structures of nine chelated YbL1-amino acid adducts (Gly, Ala, Ser, Thr, Met) confirm this. The ternary complexes with dipeptides (e.g. Gly-Ala, Gly-Ser, Gly-Met, Gly-Asp, Gly Asn, Gly-His, Ser-Met, Asp-Phe, His-Gly) also favour the terminal amine as the axial donor with the proximate amide group binding to generate a five-ring chelate. Evidence for chelation through side-chain functionality was found only in the case of N-terminal Asp. The chiral environment about the ytterbium ion upon amino acid binding has also been probed using near-IR circular dichroism spectroscopy. PMID- 15356746 TI - Synthesis and deprotonation of P-H-functionalised phosph(III)azanes; borohydride dependency on the formation of cis- or trans-[HP(mu-NtBu)2PNtBuH]. AB - The reaction of [ClP(muNtBu)2PNtBuH] (1) with LiBsBu3H yields trans [HP(muNtBu)2PNtBuH] (2), by contrast, reaction with LiBEt3H yields cis-[HP(mu NtBu)2PNtBuH] (3). Compounds and represent the first examples of P-H functionalised cyclophosph(III)azanes. Deprotonation of with BnNa (Bn=benzyl) gives the first example of a secondary phosphine-functionalised cyclodiphosph(III)azane anion [HP(mu-NtBu)2PNtBu]- (4). PMID- 15356745 TI - Effect of chelating vs. bridging coordination of chiral short-bite P-X-P (X=C, N, O) ligands in enantioselective palladium-catalysed allylic substitution reactions. AB - The chiral short-bite ligands (Ra,Ra)-bis(dinaphthylphosphonito)methane, (Ra,Ra) 1, (Ra,Ra)-bis-dinaphthylpyrophosphite, (Ra,Ra)-2, (Sc)-bis(diphenylphosphino) sec-butylamine, (Sc)-3, (Ra,Ra)-bis(dinaphthylphosphonito)phenylamine, (Ra,Ra) 4a, (Ra,Ra,Sc)-bis(dinaphthylphosphonito)-sec-butylamine, (Ra,Ra,Sc)-4b, and (Ra,Sc)-(dinaphthylphosphonito)(diphenylphosphino)-sec-butylamine, (Ra,Sc)-5, have been synthesised. The cationic palladium-allyl mononuclear chelate, [Pd(eta3 PhCHCHCHPh)(mu-L-Lshort-bite)]PF6 [L-Lshort-bite=(Sc)-3, (Ra,Ra)-4a, (Ra,Ra,Sc) 4b and (Ra,Sc)-5 for complexes, and, respectively] and binuclear bridged [Pd(eta3 PhCHCHCHPh)(mu-Ra,Ra-2)]2(PF6)2, 12, have been isolated. The short-bite chiral ligands synthesised have been tested in the palladium-allyl catalysed substitution reaction of 1,3-diphenylallyl acetate with dimethyl malonate. The catalytic system was studied, in solution, by a multinuclear NMR technique. In the catalytically active species formed with (Ra,Ra)-2 ligand, [Pd(eta3 PhCHCHCHPh)(Ra,Ra-2)]2(PF6)2, 12, the palladium(II) centres are bridged by two ligands which are forced to adopt a nearly cis-coordination to allow coordination of the allyl-moiety. Semiempirical calculations on a biphenyl-model molecule, similar to the species 12, indicate that this situation induces a strain and rigid conformation in the chiral ligands, which produce differences in the terminal allyl carbon atoms. As consequence, the catalytic product was obtained with an enantiomeric excess of 57.1% in the S form. A low e.e. value was obtained when the (Ra,Ra)-1, (Sc)-3, (Ra,Ra)-4a, (Ra,Ra,Sc)-4b and (Ra,Sc)-5 ligands have been tested in the same palladium-catalysed reaction. PMID- 15356747 TI - Stability and kinetics of the acid-promoted decomposition of Cu(II) complexes with hexaazacyclophanes: kinetic studies as a probe to detect changes in the coordination mode of the macrocycles. AB - The synthesis, protonation and Cu(II) coordination features of the novel azacyclophane type receptors 2,6,10,13,17,21-hexaza[22]-(2,6)-pyridinophane (L2), 2,6,9,12,15,19-hexaza[20]-(2,6)-pyridinophane (L5) and 2,6,9,12,15,19 hexaza[20]metacyclophane (L6) are presented. The protonation and Cu(II) constants are analysed and compared with the previously reported open-chain polyamines 4,8,11,15-tetrazaoctadecane-1,18-diamine (L1) and 4,7,10,13-tetraazahexadecane 1,16-diamine (L4) and of the cyclophane 2,6,10,13,17,21-hexaaza[22]paracyclophane (L3). All the systems form mono- and dinuclear complexes whose stability and pH range of existence depend on the type of hydrocarbon chains and molecular topology. The effects of the cyclic or open-chain nature and of the presence of the pyridine rings on the protonation and formation of mono- and dinuclear complexes are discussed. Stopped-flow kinetic measurements on the acid-promoted decomposition of the Cu(II) complexes have been carried out for the different systems. With respect to the decomposition of the dinuclear complexes, because the size of the macrocycles forces both metal ions to be close to each other, the release of the first ion occurs within the mixing time of the stopped-flow except for the dinuclear complexes of L2. However, the most interesting kinetic result is the observation of different kinetics of decomposition for the different mononuclear complexes formed by a given ligand. This effect is especially evident for L3 and L6 and indicates a change in the coordination mode of the ligand for the different mononuclear species. Therefore the Cu(II) ion performs a slippage motion through the macrocyclic cavity driven by pH changes. The stopped-flow experiments are an excellent tool to detect these slippage processes that may be present for the complexes with other macrocycles. PMID- 15356748 TI - A spectroscopic investigation of the interaction between nitrogen monoxide and copper sites of the fungal laccase from Rigidoporus lignosus. AB - The interaction of NO, with the copper centres of the laccase secreted by Rigidoporus lignosus was studied under both aerobic or anaerobic conditions. The reduction of the T1 site was always observed, as detected by the disappearance of the characteristic optical band at 604 nm (T3 presents probably the same behaviour because of the decreasing of the band at 330 nm) and the absence of its characteristic EPR signal, while T2 undergoes an initial partial and transitory reduction, its EPR signal intensity totally restoring after 24 h interaction. Different magnetic parameters of the T2 site have been detected, evidencing an increase of the hyperfine coupling constant. Furthermore, the number of superhyperfine lines on the fourth line of T2 copper was also found to increase from seven in the native to nine in the NO-treated laccase, this fact implying the coordination of a nitrogenous species to the T2 site. It was also shown that nitrite can be a source of NO, thus, paralleling the behaviour of NO-donor molecules or NO gas, but after longer interaction times. The nitrogenous species coordinated to T2 site is probably NO2-, which arises indirectly by NO oxidation. In order to understand the mechanistic pathway of this interaction, some experiments were also carried out in the presence of azide to study the interaction of NO with this laccase having its trinuclear cluster blocked by the presence of an exogenous ligand as N3-. After the addition of NO-donor molecules to the azide-treated laccase, a new EPR signal appeared at low temperatures, which is ascribable to the partially reduced T3 site, while the T1 and T2 sites were found to be totally reduced. The mechanistic pathway of the NO interaction seems to proceed through the reduction of T1 and T3 copper sites, followed by the coordination of nitrogenous species to T2. PMID- 15356749 TI - Syntheses, structural properties and catecholase activity of copper(II) complexes with reduced Schiff base N-(2-hydroxybenzyl)-amino acids. AB - A number of dicopper(II) complexes of reduced Schiff base ligands, N-(2 hydroxybenzyl)-amino acids [Cu2L2(H2O)x].yH2O (L = Sgly (1), D-Sala (2), L-Sala (3), DL-Sala (4), Sab2 (5), Sbal (6), Sab4 (7), Sval (8), Shis (9), Styr (10) and Stryp (11), x= 0-2 & y= 0-2) have been synthesized, and the solid-state structures of, and have been determined. The compounds and are binuclear in which the Cu(II) centres have square-pyramidal geometry with apical sites occupied by aqua ligands. In and one axial site is occupied by water and the other by an oxygen atom of the carboxylate group from the adjacent dimer through oxygen atoms to form 1D helical polymer. Variable temperature magnetic measurements of the dimer and helical polymer showed that they are typical for moderately strong antiferromagnetic coupling. All the complexes show significant catalytic activity on the oxidation of 3,5-di-tert-butylcatechol. The activity measured in terms of Kcat in the range 199-3800 h(-1) has been found to follow the order: 7>6>8>3>5 approximately 2 approximately 1>4 >10 >9 >11. The catalytic activity is found to increase with increasing the length of the methylene side chain of the amino acid in the reduced Schiff base ligands. PMID- 15356750 TI - Metal ion recognition via 'selective detuning'. The interaction of selected transition and post-transition metal ions with a mono-N-benzylated O2N3-donor macrocycle and its xylyl-bridged ring analogue. AB - The interaction of cobalt(II), nickel(II), copper(II), zinc(II), cadmium(II), silver(I) and lead(II) with symmetrical mono-N-benzylated and xylyl-linked macrocyclic ligands derived from the O2N3-macrocycle, 1,12,15-triaza-3,4:9,10 dibenzo-5,8-dioxacycloheptadecane, has been investigated. The log K values for the respective 1 : 1 complexes in 95% methanol (I= 0.1; Et4NClO4, 25 degrees C) of the mono-benzylated derivative have been determined potentiometrically and the results compared with the values obtained previously for the parent (non benzylated) ring system as well as for related di- and tri-benzylated macrocyclic species. Mono-benzylation results in slightly enhanced stability for the 1 : 1 silver(I) complex while the values for the corresponding complexes of the other six ions are in each case decreased even though the highest stability is still maintained for the 1 : 1 copper(II) complex. The log K results are in accord with a previous proposal that N-benzylation of amine-containing macrocyclic rings of the present type will normally have only a minor (positive or negative) influence on the affinity towards silver(I) while the corresponding binding strengths towards the remaining six metal ions are significantly reduced-behaviour we term 'selective detuning'. Competitive seven-metal transport experiments across a bulk chloroform membrane have been performed using both ligand systems as ionophores. In parallel to the log K results, transport selectivity for copper(II) was exhibited by both systems, with similar transport efficiencies being evident when compared on a 'per macrocyclic cavity' basis. PMID- 15356751 TI - Cerium(III) dialkyl dithiocarbamates from [Ce[N(SiMe3)2]3] and tetraalkylthiuram disulfides, and [Ce(kappa2-S2CNEt2)4] from the Ce(III) precursor; Tb(III) and Nd(III) analogues. AB - The synthesis and characterisation of the first neutral cerium dialkyl dithiocarbamate complexes, using a novel oxidative displacement of the amido ligands of [Ce[N(SiMe3)2]3] by tetraalkylthiuram disulfides [R2NC(S)S]2(R = Me, Et) in thf solution, are reported. In the absence of other donors, the complexes [Ce(kappa2-S2CNMe2)3(thf)2] and Ce(kappa2-S2CNEt2)3) 3 were obtained. The addition of a polypyridyl ligand allowed easy access to a range of complexes of general formula [Ce(kappa2-S2CNR2)3(L[intersection]L)][R = Me and L([intersection])L = 2,2'-bipy (4), or 4,7-diphenyl-1,10-phenanthroline (6); or R = Et and L[intersection]L = 2,2'-bipy (5)]. Brief exposure of the Ce(III) dithiocarbamate to oxygen gas afforded in high yield the diamagnetic, crystalline Ce(IV) dithiocarbamate [Ce(kappa2-S2CNEt2)4)] 7. The neodymium (8) and terbium (10) complexes, isoleptic with 2, were prepared from the appropriate 4f metal (Ln) bis(trimethylsilyl)amide [Ln[pN(SiMe3)2]3][Ln = Nd or Tb (9)] and [Me2NC(S)S]2. The structures of the crystalline complexes, 2, 4, 6, 7, 9 and 10 have been determined by X-ray crystallography. Some evidence has been obtained for the formation of the cerium(IV) complex Ce[N(SiMe3)2]2(kappa2-S2CNMe2)2. The cerium(IV) complex 7 has the metal coordinated to eight sulfur atoms of four planar chelating S2CNC2 moities and its geometry is intermediate between dodecahedral and square prismatic; the mean Ce-S bond length of 2.803 A in 7 compares with the 2.950 A in the Ce(III) complex 2. PMID- 15356752 TI - Preparative and structural studies on iron(II)-thiolate cyanocarbonyls: relevance to the [NiFe]/[Fe]-hydrogenases. AB - A number of thermally stable iron(II)-thiolate cyanocarbonyl complexes, cis,cis [Fe(CN)2(CO)2(CS3-S,S)]2-(1), mer-[Fe(CO)2(CN)3(NCCH3)]-(2)mer-[Fe(CO)3(CN)(CS3 S,S)]-(3), cis-[Fe(CO)2(CN)(S(CH2)2S(CH2)2S-S,S,S)]-(4), [Fe(CO)2(CN)3Br]2-(5), mer-[Fe(CO)2(CN)3(m-SC6H4Br)]2-(6) and mer-[Fe(CO)2(CN)3(SPh)]2-(7) were isolated and characterized by IR and X-ray diffraction analysis. The extrusion of one strong sigma-donor CN- ligand instead of CO from the iron(II) center of the thermally stable complexes [FeII(CO)2(CN)3Br]2-(5) containing less electron donating bromide reflects the electron-rich character of the mononuclear [FeII(CN)2(CO)2(CS3-S,S)]2-(1) when ligated by by the bidentate thiolate, and the combination of one cyanide, two carbonyls and a tridentate thiolate provides the stable complex 4 as a result of the reaction of complex 5 and chelating ligand [S(CH2)2S(CH2)2S]2-. The preference of the sixth ligand coordinated to the unsaturated [FeII(CO)(CN)2(CS3-S,S)]2- Fe(II) center, the iron-site architecture of the bimetallic Ni-Fe active-site of [NiFe] hydrogenases, is a strong pi acceptor CO group. Scrutiny of the coordination chemistry of iron(II)-thiolate cyanocarbonyl species [FeII(CO)x(CN)y(SR)z]n- reveals that certain combinations of thiolate, cyanide and carbonyl ligands (3 < or = y+z > or = 4) bound to Fe(II) are stable and this could point the way to understand the reasons for Nature's choice of combinations of these ligands in hydrogenases. PMID- 15356753 TI - Syntheses, structures and properties of 1-ethyl-3-methylimidazolium salts of fluorocomplex anions. AB - Fluoroacid-base reactions of a room-temperature ionic liquid, 1-ethyl-3 methylimidazolium fluorohydrogenate (EMIm(HF)2.3F, EMIm = 1-ethyl-3 methylimidazolium cation), and Lewis fluoroacids (BF3, PF5, AsF5, NbF5, TaF5 and WF6) give EMIm salts of the corresponding fluorocomplex anions, EMImBF4, EMImPF6, EMImAsF6, EMImNbF6, EMImTaF6 and EMImWF7, respectively. Attempts to prepare EMImVF6 by both the acid-base reaction of EMIm(HF)2.3F with VF5 and the metathesis of EMImCl with KVF6 failed due to the strong oxidizing power of the pentavalent vanadium, whereas EMImSbF6 was successfully prepared only by the metathesis of EMImCl and KSbF6. EMImBF4, EMImSbF6, EMImNbF6, EMImTaF6 and EMImWF7 are liquids at room temperature whereas EMImPF6 and EMImAsF6 melts at around 330 K. Raman spectra of the obtained salts showed the existence of the EMIm cation and corresponding fluorocomplex anions. IR spectroscopy revealed that strong hydrogen bonds are not observed in these salts. EMImAsF6(mp 326 K) and EMImSbF6(mp 283 K) are isostructural with the previously reported EMImPF6. The melting point of the hexafluorocomplex EMIm salt decreases with the increase of the size of the anion (PF6- < AsF6- < SbF6- 2 >3. L1-3 and undergo cleavage of the tert-butylamide upon addition of Me4NOH.5H2O (1 equiv.) in methanol at 50(1)degrees C. The rate of amide cleavage follows the order 1> 2>> 3, L1-3. The extent by which the amide cleavage reaction is accelerated in 1-3 relative to the free ligands, L1-3, is correlated with the strength of amide oxygen binding and Lewis acidity of the zinc(II) centre in deduced from the X-ray, NMR and IR studies. PMID- 15356758 TI - Coordination chemistry of 2-(8-aminoquinolino)-4,6-di-tert-butylphenol with manganese(IV), iron(III), and cobalt(II/III):N,O-coordinated o iminobenzosemiquinonate(1-) pi radical monoanions vs.o-iminophenolate(2-) dianions. AB - The potentially tridentate ligand 2-(8-aminoquinolino)-4,6-di-tert-butylphenol, H[1LAP], has been synthesized and its coordination chemistry with Mn(IV), Fe(III), and Co(III) has been investigated by X-ray crystallography, electro- and magnetochemistry, electronic, Mossbauer and EPR spectroscopies. The following complexes have been prepared [MnIV(1LAP-H)2](1), St=3/2; [MnIV(1LAP H)(1LISQ)](PF6).CH2Cl2 (2), St= 1; [FeIII(1LISQ)2](ClO4).0.5H2O (3), St=1/2; [FeIII(1LISQ)(3,5-dtcat)]2 (4), St= 0; K[CoII(1LISQ)(1LIBQ)](NCS)2 (5), St= 1; [CoIII2(NCS)2(1LAP-H)2(AQ)] (6), St=0, where (1LAP-H)2- corresponds to the o iminophenolate(2-) dianion, (1LISQ)- is the o-iminobenzosemiquinonate(1-) pi radical derivative of the ligand H[1LAP], (1LIBQ)0 is the neutral iminobenzoquinone ligand, and 3,5-dtcat is 3,5-di-tert-butylcatecholate(2-) and N,N-coordinated (AQ) is 8-aminoquinoline. It is shown that the pi radical anions in 2, 3, 4 couple antiferromagnetically to the respective paramagnetic metal ion. Complex 4 is a dinuclear neutral complex with weak antiferromagnetic coupling between two [FeIII1LISQ)(3,5-dtcat)] halves. The asymmetrically ligated complex 6 contains two bridging (1LAP-H)2- ligands and two diamagnetic CoIII ions. In contrast, 5 is correctly described as [CoII(1LISQ)(1LIBQ)]+ rather than [CoIII(1LISQ)2]+ since it possesses a temperature-independent magnetic moment of mueff(10-298 K)= 3.0 muB indicating an St=1 ground state which is attained via strong antiferromagnetic coupling (|J> or =200 cm(-1)) between a high spin cobalt(II) ion (SCo=3/2) and a single pi radical anion (Srad=1/2). PMID- 15356760 TI - Prospective evaluation of outcome measures in free-flap surgery. AB - Free-flap failure is usually caused by venous or arterial thrombosis. In many cases, lack of experience and surgical delay also contribute to flap loss. The authors prospectively analyzed the outcome of 57 free flaps over a 28-month period (January, 1999 to April, 2001). The setting was a university hospital tertiary referral center. Anastomotic technique, ischemia time, choice of anticoagulant, and the grade of surgeon were recorded. The type of flap, medications, and co-morbidities, including preoperative radiotherapy, were also documented. Ten flaps were re-explored (17 percent). There were four cases of complete flap failure (6.7 percent) and five cases of partial failure (8.5 percent). In patients who received perioperative systemic heparin or dextran, there was no evidence of flap failure (p = .08). The mean ischemia time was similar in flaps that failed (95 +/- 29 min) and in those that survived (92 +/- 34 min). Also, the number of anastomoses performed by trainees in flaps that failed (22 percent), was similar to the number in flaps that survived (28 percent). Nine patients received preoperative radiotherapy, and there was complete flap survival in each case. This study reveals that closely supervised anastomoses performed by trainees may have a similar outcome to those performed by more senior surgeons. There was no adverse effect from radiotherapy or increased ischemia time on flap survival. PMID- 15356762 TI - Reconstruction of a partial mastectomy defect with the superficial inferior epigastric artery (SIEA) flap. AB - A case in which a superficial inferior epigastric artery (SIEA) flap was used to reconstruct a partial mastectomy defect is described. The SIEA vessels have been shown to be predictable and to match well with the recipient internal mammary vessels. In addition, this flap does not require opening of the anterior rectus sheath, and it is possible to design a smaller flap, which would allow closure at the donor site without translocation of the umbilicus. The risk of developing an abdominal hernia or bulge is therefore nonexistent, and a very low and more inconspicuous scar is produced, both of which are advantages for the patient. PMID- 15356764 TI - Symptomatic progression of congenital ulnar nerve hypoplasia with brachial plexus anomaly. AB - Symptomatic progression of congenital ulnar nerve hypoplasia in association with an anomaly of the brachial plexus is uncommon. The authors present such a case involving an 11-year-old girl, who presented with a complaint of progressive numbness and hand weakness occurring in less than 6 months' time. Physical examination revealed limb hypoplasia, neurologic abnormalities, and a palpable hypoplastic ulnar nerve at the level of the cubital tunnel. An additional intraoperative finding was an anomalous brachial plexus. This combined case of congenital ulnar nerve hypoplasia, brachial plexus anomaly, and symptomatic progression, is discussed in the context of other literature. PMID- 15356765 TI - Simultaneous reconstruction of large maxillary and mandibular defects with a fibular osteocutaneous flap combined with an anterolateral thigh flap. AB - This article reports the simultaneous reconstruction of maxillary and mandibular defects caused by a close-range gunshot blast to the face with one fibular osteocutaneous flap combined with an anteroateral fasciocutaneous flap. A fibular osteocutaneous flap was used for both mandibular and maxillary defects, using multiple osteotomies and discarding a central bony segment and an oral floor defect. An anterolateral thigh flap was used to cover a three-dimensional defect of both the intraoral mucosal region, as well as external skin and soft tissue defects, including some on the upper and lower lips. The results demonstrated that the method was a good choice in the reconstruction of large composite facial defects, both aesthetically and functionally. PMID- 15356766 TI - Invited discussion: "New approach to vascular injection in fresh cadaver dissection" (J Reconstr Microsurg 2004;20:311-315). PMID- 15356767 TI - A simple method for the control of medicinal leeches. AB - The medicinal leech, Hirudo medicinalis, has been widely used in the salvage of microvascular free flaps. Numerous publications have detailed the biology, use, benefits, and risks of leech therapy. One reported significant risk is the risk of leech movement or migration from the surgical site, possibly into body orifices or even deeper into the wound itself. The authors report a simple method of limiting the movement of medicinal leeches from the surgical site, namely, affixing one end of a surgical suture to the leech and tying the free end to a firm object or dressing. This simple method limits the potential range of movement of the leech and reduces the risk of leech migration to unwanted areas. PMID- 15356768 TI - Selective transfer of the C7 nerve root: an experimental study. AB - The authors present selective C7 nerve root transfer in a rat model. The musculocutaneous nerve was neurotized by various portions of ipsilateral C7. The latent period and maximum amplitude of evoked motor action potential of the biceps, number of regenerating myelinated nerve fibers, cross-sectional area and wet weight of the biceps, and twitch and tetanic tensions of the biceps were measured at four postoperative intervals. In the early postoperative period (1 and 2 months), nerve regeneration in neurotization with the posterior division or the anterior division of C7 was significantly better than that with the anterolateral fascicles of the anterior division or the phrenic nerve. As the postoperative interval prolonged, the parameters of nerve regeneration in the latter two groups approximated those in the former two groups. This indicated that there were enough regenerating nerve fibers in the anterolateral fascicles of the anterior division and a promising potential for nerve regeneration. The clinical significance of the results lies in the design of selective C7 transfer which, using the anterolateral fascicles of the anterior division, could preserve the function of the muscles innervated by the posterior division to the greatest extent, and provide sufficient donor outflow as well. It is therefore a new option for C7 transfer. PMID- 15356770 TI - Regeneration along intact nerves using nerve growth factor and ciliary neurotrophic factor. AB - The purpose of this investigation was to evaluate the ability of a specific growth factor combination, nerve growth factor (NGF) and ciliary neurotrophic factor (CNTF), to enhance peripheral nerve regeneration. Eight groups of eight Sprague-Dawley rats underwent repair of a nerve gap defect: Group A (immediate repair), Group B (intact nerve bridge), Group C (nerve autograft), Group D (gap in situ), Group E (NGF + CNTF), Group F (NGF), Group G (CNTF), and Group H (saline). Twelve weeks after surgery, analysis included the measurement of the isometric force of muscle contraction for the tibialis anterior muscle and tissue harvesting for both quantitative and qualitative analysis. When evaluating muscle contraction force, there was no statistically significant difference among the experimental groups receiving a growth factor injection and the normal saline control group. The hypothesis of this study was that peripheral nerve regeneration could be enhanced by the combination of NGF and CNTF. The evidence does not support this hypothesis. PMID- 15356771 TI - Muscle flap mass preservation with end-to-side neurorrhaphy: an experimental study. AB - The authors examined the preservation of rat gracilis muscle flap mass after motor and sensory end-to-side neurorrhaphy. The rat gracilis muscle flap model was designed based on a previous study. Twenty-four Sprague-Dawley rats were divided into three groups. In Group 1 (n = 8), the flap was denervated by transecting the obturator nerve. In Group 2 (n = 8), the flap was reinnervated by coapting the proximal saphenous nerve to the distal obturator nerve. In Group 3 (n = 8), the flap was reinnervated by coapting the motor branch of the femoral nerve to the distal stump of the obturator nerve. At 6 months postoperatively, the gracilis muscle flaps were examined, harvested, and weighed individually. Results showed that the flaps with motor nerve reinnervation retained good bulk, with a weight of 634.0 +/- 65.1 gm, which was statistically significantly higher than the denervated group (457.5 +/- 125.3 gm, p < 0.01). However, muscle mass preservation in the sensory reinnervated group (606.9 +/- 209.1 gm) was not significantly different, compared to the denervated group. Histology revealed atrophic changes in the denervated group, compared to the sensory and motor reinnervated groups. The authors concluded that muscle mass can be preserved by end-to-side nerve repair. Motor nerve reinnervation is able to better arrest atrophic changes of the muscle flaps. PMID- 15356773 TI - Primary care guidelines for the management of persons infected with human immunodeficiency virus: recommendations of the HIV Medicine Association of the Infectious Diseases Society of America. PMID- 15356774 TI - Multiple-year experience in the diagnosis of viral central nervous system infections with a panel of polymerase chain reaction assays for detection of 11 viruses. AB - BACKGROUND: Polymerase chain reaction (PCR) is becoming more common in diagnostic laboratories. In some instances, its value has been established. In other cases, assays exist, but their beneficial use has not been determined. This article summarizes findings from 3485 patients who underwent testing over a 6-year period in our laboratory. METHODS: A panel of PCR assays was used for the detection of a range of viruses associated with central nervous system (CNS) infections. PCR results were analyzed in conjunction with information about patient age and sex, the time between onset and specimen collection, and other variables. Medical chart review was conducted for 280 patients to gain diagnostic and epidemiologic insight with regard to cases of unresolved encephalitis. RESULTS: A total of 498 PCR-positive samples (14.3%) were detected. Enteroviruses accounted for the largest number (360 [72.3%]) of positive PCR results, followed by herpes simplex virus (76 [15.3%]), varicella-zoster virus (29 [5.82%]), and West Nile virus (WNV) (18 [3.61%]). Of 360 patients who tested positive for enterovirus, only 46 met the Centers for Disease Control and Prevention's encephalitis definition. It resulted in the greatest decrease (87.2%) in positive PCR results. Overall, the PCR positivity rate for specimens collected within 5 days after illness onset was 17.2%, compared with 8.6% for specimens collected > or =6 days after onset. CONCLUSIONS: The value of PCR in the diagnosis of viral infections has been established. PCR is of lower value in the detection of WNV in CNS, compared with serological testing, but is of greater value in the detection of other arboviruses, particularly viruses in the California serogroup. Medical chart reviews indicated that apparent CNS infection resolves in approximately 50% of cases. PMID- 15356775 TI - Presence of viremia in patients with primary herpetic gingivostomatitis. AB - BACKGROUND: Presence of viremia during primary herpes simplex virus (HSV) infections has been previously investigated, but the findings for immunocompetent individuals have only rarely been reported. METHODS: With use of polymerase chain reaction (PCR), we evaluated blood samples obtained from children with primary herpes simplex virus (HSV) gingivostomatitis for viremia. RESULTS: There were 16 girls and 16 boys, aged 9-44 months (median age, 19 months). Serological test results for HSV type 1 were positive for 3 subjects (10.3%), borderline for 7 (24.1%), and negative for 19 (65.5%). Results of PCR of peripheral blood samples were positive for 11 subjects (34.4%). Time from disease onset to specimen collection was 24-216 h (median, 72 h) and was longer for subjects with positive results of serological tests (P =.014) and shorter for subjects with positive PCR results (P=.42). No cases with positive results of both PCR and serological tests were found. CONCLUSION: PCR detected viremia in 34% of patients with primary herpetic gingivostomatitis. Presence of viremia may play a potential role in viral dissemination, providing a better understanding of the pathogenesis of HSV infections, especially of the central nervous system. PMID- 15356776 TI - Reduction in high rates of antibiotic-nonsusceptible invasive pneumococcal disease in tennessee after introduction of the pneumococcal conjugate vaccine. AB - BACKGROUND: Invasive pneumococcal disease (IPD) is a burgeoning problem, with rates of antibiotic-nonsusceptible IPD, in particular, increasing during the past decade. One measure to combat IPD is vaccination with the recently introduced 7 valent pneumococcal conjugate vaccine (PCV). METHODS: To evaluate the effects of the introduction of PCV in 2000 on the epidemiology of antibiotic-nonsusceptible IPD, a database of IPD cases from January 1995 through December 2002 identified through active surveillance in 5 Tennessee counties was examined. For each case, clinical data were collected, and antibiotic susceptibility testing and serotyping were performed on available isolates. RESULTS: Among children younger than 2 years, IPD rates peaked at 235 cases per 100,000 in 1999 before decreasing, after PCV licensure, to 46 cases per 100,000 in 2002 (P<.001). The proportion of penicillin-nonsusceptible IPD isolates from this age group declined from 59.8% in 1999 to 30.4% in 2002 (P<.01). After 2001, similar decreases in IPD rates and in the proportion of antibiotic-nonsusceptible isolates recovered were seen among persons aged 2 years and older (P<.01). Rates of IPD due to PCV associated serotypes declined after PCV introduction in all age groups (P<.001), whereas the rate of IPD due to nonvaccine serotypes increased among persons aged 2 years and older. CONCLUSIONS: In the 2 years since licensure, widespread PCV vaccination of children has resulted in dramatic declines in the proportion of antibiotic-nonsusceptible isolates in Tennessee. PCV vaccination of children also appears to be a highly effective method for reducing the burden of IPD in adults. PMID- 15356777 TI - Vaccination: a novel approach to reduce antibiotic resistance. PMID- 15356778 TI - Severe acute respiratory syndrome coronavirus on hospital surfaces. AB - BACKGROUND: Health care workers continued to contract severe acute respiratory syndrome (SARS), even after barrier precautions were widely implemented. METHODS: We explored the possible contribution of contaminated hospital surfaces to SARS transmission by swabbing surfaces in 2 hospitals and testing the swab samples by reverse-transcriptase polymerase chain reaction (RT-PCR) and viral culture. RESULTS: Twenty-six of 94 swab samples tested positive for viral RNA. Swab samples of respiratory secretions from each of the 4 patients examined tested positive by RT-PCR, as were 12 of 43 swabs from patient rooms and 10 of 47 swabs from other parts of the hospital, including the computer mouses at 2 nursing stations and the handrail of the public elevator. Specimens from areas with patients with SARS in the most infectious phase of illness (days 5-15 after onset) were more likely to be RNA positive than were swab specimens from elsewhere (24 of 63 samples vs. 2 of 31 samples; P=.001). All cultures showed no growth. CONCLUSIONS: Although the viruses identified may have been noninfectious, health care workers should be aware that SARS coronavirus can contaminate environmental surfaces in the hospital, and fomites should be considered to be a possible mode of transmission of SARS. PMID- 15356779 TI - A prospective, randomized trial of 3 or 14 days of ciprofloxacin treatment for acute urinary tract infection in patients with spinal cord injury. AB - BACKGROUND: Urinary tract infection (UTI) is common among patients with spinal cord injury. The optimal duration of treatment for symptomatic UTI has not been determined. METHODS: A randomized, double-blind, placebo-controlled trial compared 3-day and 14-day regimens of ciprofloxacin, 250 mg twice daily, for the treatment of acute UTI in patients with spinal cord injury. Patients with pyelonephritis, struvite stones, hydronephrosis, or long-term indwelling catheters were excluded from the trial. RESULTS: Sixty patients with spinal cord injury were enrolled in the trial, with 30 patients assigned to each study arm. The most common infecting organisms were Klebsiella species (30%), Enterococcus species (22%), and Escherichia coli (22%); 33% of the infections were polymicrobial. Microbiological cure at long-term follow-up was significantly better among patients who received therapy for 14 days than among patients who received therapy for 3 days. By 6 weeks of follow-up, microbiological relapse (in 11 [37%] of 30 patients vs. 2 [7%] of 30 patients; 95% confidence interval [CI], 1.38-3.18; P=.01) and symptomatic relapse (in 7 [23%] 30 patients vs. 0 of 30 patients; 95% CI, 1.69-3.13; P=.01) both occurred more frequently in patients treated for 3 days. Reinfection occurred with similar frequency in patients in the 2 study arms. Six of 7 evaluable patients with treatment failure had a fluoroquinolone-resistant organism isolated at enrollment. CONCLUSIONS: For patients with spinal cord injury, treatment of acute symptomatic UTI for 14 days leads to improved clinical and microbiological outcomes, compared with short course therapy. PMID- 15356780 TI - Duration of therapy for urinary tract infection: the long and the short of it. PMID- 15356781 TI - Health care workers and the initiation of treatment for latent tuberculosis infection. AB - BACKGROUND: Despite strong evidence and recommendations supporting the treatment of latent tuberculosis infection, many affected health care workers at risk of acquiring tuberculosis from and potentially transmitting tuberculosis to their patients do not receive treatment. The objective of this study was to determine whether health care workers were less likely than non-health care workers to initiate treatment for latent tuberculosis infection. METHODS: In this retrospective cohort study that used the disease management database from a specialized downtown Toronto tuberculosis clinic, patients with latent tuberculosis infection were included if they had risk factors for progression of disease and were excluded if they had contraindications to treatment. RESULTS: Our final cohort consisted of 308 patients with latent tuberculosis infection. The overall treatment initiation rate was 58%. We found that, when a number of confounding variables, including age, foreign birth, contact with persons with active tuberculosis, tuberculosis skin test conversion, bacille Calmette-Guerin vaccination, abnormal chest radiograph findings, comorbidities, and income, were considered and/or controlled for, the odds of a health care worker initiating treatment were approximately one-half of those of a non-health care worker (adjusted odds ratio, 0.55; 95% confidence interval, 0.32-0.93). CONCLUSION: We conclude that, in our clinic, health care workers are less likely than non-health care workers to initiate treatment for latent tuberculosis infection. PMID- 15356782 TI - Pretransplantation respiratory syncytial virus infection: impact of a strategy to delay transplantation. AB - BACKGROUND: Delay of hematopoietic stem cell transplantation (HSCT) has been suggested if upper respiratory tract infection (URTI) due to respiratory syncytial virus (RSV) occurs in transplantation candidates, but the efficacy of this strategy in preventing posttransplantation RSV infection is unknown. METHODS: In a retrospective study, we reviewed charts of patients who underwent transplantation at Fred Hutchinson Cancer Research Center (Seattle, WA) during the period of June 1987 through December 2000 and evaluated the strategy of delaying HSCT in candidates with laboratory-confirmed RSV URTI. RESULTS: Thirty one of 37 patients had RSV URTI before conditioning, 2 (6.5%) of whom developed RSV infection after HSCT. In 6 of 37 patients, symptoms of URTI were present during the start of conditioning, but RSV virologic confirmation occurred a median of 4.5 days (range, 2-5 days) into the conditioning regimen. Conditioning was aborted for 3 of 6 patients; none had progression to RSV pneumonia. Of the 3 patients in whom HSCT proceeded as scheduled, 2 developed RSV pneumonia. Overall, RSV pneumonia occurred in 1 of 34 patients for whom HSCT was delayed, compared with 2 of 3 patients for whom there was no delay (P=.01). CONCLUSIONS: In patients with pretransplantation RSV URTI, delay of HSCT was associated with a lower risk of pneumonia than was no delay. Because URTIs can progress to severe complications in patients receiving HSCTs, these results support Centers for Disease Control and Prevention/American Society of Blood and Marrow Transplantation recommendations that HSCT be delayed on the basis of symptoms of URTI rather than waiting for virologic confirmation. PMID- 15356783 TI - Etiology of community-acquired pneumonia in hospitalized school-age children: evidence for high prevalence of viral infections. AB - BACKGROUND: Community-acquired pneumonia (CAP) in young children is most commonly associated with viral infections; however, the role of viruses in CAP of school age children is still inconclusive. METHODS: Seventy-five school-age children hospitalized with CAP were prospectively evaluated for the presence of viral and bacterial pathogens. Nasopharyngeal washes were examined by polymerase chain reaction for viruses and atypical bacteria. Antibody assays to detect bacterial pathogens in acute-phase and convalescent-phase serum samples were also performed. RESULTS: A viral infection was identified in 65% of cases. Rhinovirus RNA was detected in 45% of patients; infection with another virus occurred in 31%. The most common bacterial pathogen was Mycoplasma pneumoniae, which was diagnosed in 35% of cases. Chlamydia pneumoniae DNA was not detected in any patient; results of serological tests were positive in only 2 patients (3%). Mixed infections were documented in 35% of patients, and the majority were a viral-bacterial combination. CONCLUSIONS: The high prevalence of viral and mixed viral-bacterial infections supports the notion that the presence of a virus, acting either as a direct or an indirect pathogen, may be the rule rather than the exception in the development of CAP in school-age children requiring hospitalization. PMID- 15356784 TI - Reptile-associated salmonellosis in preschool-aged children in Michigan, January 2001-June 2003. AB - OBJECTIVES: Determine the incidence of reptile-associated salmonellosis in preschool-aged children in Michigan. METHODS: Cases of reptile-associated salmonellosis in children < or =5 years of age occurring in Michigan January 2001 June 2003 were identified through review of individual patient case-history forms provided by local health departments to the Michigan Department of Community Health and by identification of Michigan Department of Community Health laboratory-confirmed cultures of reptile-associated serotypes, determined by evaluation of the Public Health Laboratory Information System's Clinical Nonhuman Salmonella data for 1990-2001. RESULTS: The incidence of reptile-associated salmonellosis was 11.8% of all Salmonella cases reported in Michigan children aged < or =5 years for the period January 2001 through June 2003. CONCLUSIONS: Despite the recommendations of the Centers for Disease Control and Prevention to avoid the exposure of children <5 years old to reptiles, reptile-associated salmonellosis in preschool-aged children continues to be a public health problem in Michigan. PMID- 15356785 TI - Long-term interferon-gamma therapy for patients with chronic granulomatous disease. AB - BACKGROUND: Chronic granulomatous disease (CGD) is a rare disorder of phagocytes in which absent production of superoxide and hydrogen peroxide in phagocytes predisposes patients to bacterial and fungal infections. Infections are dramatically reduced by prophylaxis with antibiotics, antifungals, and interferon gamma (IFN-gamma ). METHODS: Seventy-six patients with CGD were enrolled in an uncontrolled, open-label follow-up study to assess the long-term clinical safety and efficacy of IFN-gamma therapy. Patients received IFN-gamma subcutaneously 3 times per week. RESULTS: We observed patients for up to 9 years, for a total observation period of 328.4 patient-years. The incidence of serious infections was 0.30 infections per patient-year; for serious bacterial infections, the incidence was 0.18 cases per patient-year, and for serious fungal infections, it was 0.12 cases per patient-year. Thirty-seven percent of patients reported an adverse event, the most common of which was fever. Twenty-six patients withdrew from the study (3 because of adverse events, 15 because of patient preference, and 8 because of transfer to another trial). There were no life-threatening IFN gamma-related adverse events and no discernible effects on growth. The overall mortality rate was 1.5% per patient-year. CONCLUSION: IFN-gamma prophylaxis for CGD appears to be effective and well tolerated over a prolonged period of time. PMID- 15356786 TI - Disinfection and sterilization in health care facilities: what clinicians need to know. AB - All invasive procedures involve contact between a medical device or surgical instrument and a patient's sterile tissue or mucous membranes. A major risk of all such procedures is the introduction of pathogenic microbes that could lead to infection. Failure to properly disinfect or sterilize reusable medical equipment carries a risk associated with breach of the host barriers. The level of disinfection or sterilization is dependent on the intended use of the object: critical items (such as surgical instruments, which contact sterile tissue), semicritical items (such as endoscopes, which contact mucous membranes), and noncritical items (such as stethoscopes, which contact only intact skin) require sterilization, high-level disinfection, and low-level disinfection, respectively. Cleaning must always precede high-level disinfection and sterilization. Users must consider the advantages and disadvantages of specific methods when choosing a disinfection or sterilization process. Adherence to these recommendations should improve disinfection and sterilization practices in health care facilities, thereby reducing infections associated with contaminated patient-care items. PMID- 15356787 TI - Mitochondrial effects of HIV infection on the peripheral blood mononuclear cells of HIV-infected patients who were never treated with antiretrovirals. AB - To investigate the effects of HIV infection on mitochondrial DNA (mtDNA) content and other mitochondrial parameters, we used peripheral blood mononuclear cells (PBMCs) from 25 asymptomatic antiretroviral-naive human immunodeficiency virus (HIV)-infected patients and from 25 healthy control subjects. HIV-infected patients had significant decreases in mtDNA content (decrease, 23%; P<.05) and in the activities of mitochondrial respiratory chain (MRC) complex II (decrease, 41%; P<.001), MRC complex III (decrease, 38%; P<.001), MRC complex IV (decrease, 19%; P=.001), and glycerol-3-phosphate dehydrogenase (decrease, 22%; P<.001), along with increased lipid peroxidation of PBMC membranes (P=.007). Therefore, HIV infection is associated not only with mtDNA depletion, but also with extensive MRC disturbances and increased oxidative damage. PMID- 15356788 TI - Prevalence of clinical symptoms associated with highly active antiretroviral therapy in the Women's Interagency HIV Study. AB - BACKGROUND: The extended use of antiretroviral drugs among human immunodeficiency virus (HIV)-seropositive individuals underscores the need for a comprehensive evaluation of therapy-associated clinical symptoms. METHODS: Beginning in April 2000, 364 HIV-seronegative and 1256 HIV-seropositive women enrolled in a multicenter cohort study reported clinical symptoms that included abdominal pain, diarrhea, anorexia, nausea and/or vomiting, myalgias, fatigue, fever, body fat redistribution, dizziness, headaches, paresthesias, xerostomia, nephrolithiasis, and rash. We examined the prevalence of symptoms with respect to HIV infection and the use of highly active antiretroviral therapy (HAART), using data correlation models. RESULTS: In the 6 months before a study visit, 49% of HIV seronegative women, 67% of HIV-seropositive women not receiving therapy, and 69% of HIV-seropositive women receiving HAART reported any clinical symptom. The odds ratios (ORs) for reporting any symptom were 1.4 (95% confidence interval [CI], 1.1-1.8) for women who changed HAART regimens and 0.9 (95% CI, 0.7-1.1) for women reporting stable HAART use, compared with those reporting no therapy use. Significant findings (P<.05) for particular symptoms were an increased odds of diarrhea, nausea and/or vomiting, body fat redistribution, myalgias, and paresthesias, when data for women who changed HAART regimens were compared with those for women not receiving therapy. The OR for reporting any symptom was 1.5 (95% CI, 1.2-1.9) for women who switched HAART regimens and 1.6 (95% CI, 1.3-1.9) for women who discontinued HAART, compared with those reporting stable HAART use. CONCLUSIONS: Our findings confirm the high prevalence of clinical symptoms among HIV-seropositive women who changed HAART regimens. The high prevalence of symptoms among HIV-seronegative women and HIV-seropositive women not receiving therapy demonstrates that caution should be used when attributing the occurrence of symptoms entirely to HAART. PMID- 15356789 TI - Decrease in hospitalization and mortality rates among children with perinatally acquired HIV type 1 infection receiving highly active antiretroviral therapy. AB - BACKGROUND: The impact of highly active antiretroviral therapy (HAART) on human immunodeficiency virus type 1 (HIV-1) disease progression in perinatally infected children is not well documented. This study aims to identify the effect of evolving antiretroviral therapy on the immunologic and virologic status of and hospitalization and mortality rates among perinatally infected children. METHODS: Children receiving outpatient care during 1994-2001 at 3 HIV clinics in southern California were observed longitudinally for CD4+ cell percentage, plasma HIV-1 RNA load, antiretroviral treatment, Pneumocystis jiroveci pneumonia (PCP) prophylaxis, and rate of hospital admissions. RESULTS: A total of 129 children were observed during the study period; 51% were girls, and 40.3% were Hispanic, 29.5% were African American, and 27.1% were white. The mean CD4+ cell percentage increased from 22.5% in 1994 to 31.2% in 2001 (P<.01), and the mean plasma HIV-1 RNA load decreased from 4.53 log10 copies/mL in 1996 to 3.27 log10 copies/mL in 2001 (P<.001). The use of HAART increased from 0% in 1994 to 93% in 2001 (P<.01), whereas the use of PCP prophylaxis decreased from 55% to 16% during this time (P<.001). The hospitalization rate decreased from 6.49 to 0.60 admissions per 100 person-years during 1994-2001 (P<.001). Acquired immunodeficiency syndrome associated hospital admission rates decreased from 15.6% in 1994 to 0% in 2001 (P<.0001). Similarly, the admission rate for patients with Centers for Disease Control and Prevention category B decreased from 29.7% in 1994 to 5.9% in 2001 (P<.0001). Logistic regression analysis showed that CD4+ cell percentage and viral load were independently associated with risk of hospitalization. Survival was significantly longer for those who received HAART. CONCLUSIONS: HIV-1 associated mortality and hospitalization rates decreased significantly between 1994 and 2001 in perinatally infected children. This correlated with an increase in CD4+ cell percentage and a decrease in HIV-1 RNA load concurrently with the expanded use of HAART. PMID- 15356790 TI - The effects of low-dose growth hormone in HIV-infected men with fat accumulation: a pilot study. AB - Pharmacologic doses of growth hormone (GH) reduce HIV-associated fat accumulation but may worsen glucose metabolism. We investigated the effects of a low dose of GH (1 mg per day) in HIV-infected men with fat accumulation and found that such treatment reduced total fat and increased lean body mass without significant changes in glucose tolerance or insulin sensitivity. Visceral adipose tissue (VAT) levels did not change significantly for the group as a whole, although a reduction in the VAT level was seen in patients with a greater VAT level at baseline. PMID- 15356791 TI - Nelfinavir plasma concentrations are low during pregnancy. AB - Plasma nelfinavir concentration ratios (CRs) were calculated for all pregnant (n=27) and nonpregnant (n=48) human immunodeficiency virus type 1-infected women receiving the drug who visited our outpatient clinic. In pregnant women, mean and median nelfinavir CRs were significantly lower (P=.02 and P=.04, respectively), and 51% of the CRs were below the clinically relevant threshold of 0.90, compared with 35% of the CRs in nonpregnant women. After we adjusted for confounders, we found that the mean nelfinavir CR was 34% lower in pregnant women (P=.02). With targeted interventions, subsequent CRs in pregnant women showed a significant increase (median increase, 0.31; P=.01). PMID- 15356792 TI - Breakthrough fungal infections in stem cell transplant recipients receiving voriconazole. AB - Infection with voriconazole-resistant fungi may become problematic, because organisms with decreased susceptibility have been noted. Breakthrough fungal infections occurred in 13 of 139 patients who received voriconazole at our center during the period of September 1998 through September 2003. Zygomycetes were found in 6 patients, and Candida glabrata bloodstream infection occurred in 4 patients. Minimal inhibitory concentrations were > or =1 microg/mL for all available isolates. Yeasts and molds with decreased susceptibility to voriconazole may cause invasive infection in patients treated successfully for aspergillosis. PMID- 15356793 TI - Staphylococcus aureus and Clostridium difficile cause distinct pseudomembranous intestinal diseases. AB - We report simultaneous infections with Clostridium difficile and methicillin resistant Staphylococcus aureus (MRSA) in a patient with discrete colonic pseudomembranes typical of C. difficile infection, as well as confluent, loosely adherent pseudomembranes in the small bowel. Identification of MRSA in the small bowel pseudomembrane by polymerase chain reaction supports S. aureus as an enteric pathogen. PMID- 15356794 TI - Risk factors for leptospirosis in metropolitan France: results of a national case control study, 1999-2000. AB - Risk factors for leptospirosis in France were investigated to improve the vaccination program for this disease. Data from 90 hospitalized case patients and 169 matched control subjects were analyzed in a case-control study. Skin lesions, canoeing, contact with wild rodents, and country residence were independently associated with leptospirosis, emphasizing that leisure activity is a risk factor for this illness. PMID- 15356796 TI - Intranasal versus injectable influenza vaccine. PMID- 15356797 TI - Clinical relevance of bacteriostatic versus bactericidal activity in the treatment of gram-positive bacterial infections. PMID- 15356798 TI - Implementation of an antibiotic control program. PMID- 15356799 TI - Mycotic hepatic artery aneurysm complicating Staphylococcus aureus endocarditis: successful diagnosis and treatment. PMID- 15356800 TI - New times for an old disease: intracranial mass lesions caused by Mycobacterium tuberculosis in 5 HIV-negative African immigrants. AB - BACKGROUND: The tuberculosis epidemic is still a global emergency, and its spread in the past 20 years has been fueled by the acquired immune deficiency syndrome pandemic and increasing drug resistance. International travel and migration may increase the incidence of tuberculosis in industrialized countries. METHODS: We reviewed the clinical charts of patients admitted to the infectious diseases unit of Ospedali Riuniti (Bergamo, Italy) to identify patients with intracranial mass lesions caused by Mycobacterium tuberculosis. RESULTS: During the past 6.5 years, 5 of 30 patients with a mass of infectious origin in the brain had tuberculous brain lesions diagnosed. All 5 were human immunodeficiency virus (HIV)-negative adults and African immigrants. No patient had concomitant meningitis, 1 had a concomitant pulmonary disease, and 3 subjects reported a past history of tuberculosis. At presentation, no patient had fever and 3 had seizures. Examination of cerebrospinal fluid revealed normal findings for 4 of 4 subjects, and neuroimaging showed multiple intracranial mass lesions in 4 of 5 patients. The diagnosis was definite for 2 subjects (based on analysis of brain specimens) and presumptive for 3 subjects (1 had concomitant pulmonary tuberculosis, and 2 had clinical response to therapy). Results of susceptibility tests for M. tuberculosis were available for 2 patients: both isolates were resistant to isoniazid, and 1 was also resistant to streptomycin. Duration of medical treatment ranged from 11 to 23 months, and 2 subjects underwent surgical procedures at the time of diagnosis. All 5 patients recovered. CONCLUSIONS: Clinicians in western countries should consider the possible role of tuberculosis in causing mass lesions in the brain, particularly in immigrants from regions where tuberculosis is endemic. PMID- 15356801 TI - Shiga toxin-producing Escherichia coli as a possible etiological agent of chronic diarrhea. AB - We identified Shiga toxin-producing Escherichia coli (STEC) as the likely etiologic pathogen for chronic diarrhea in 2 patients, 1 of whom was immunocompromised with acquired immunodeficiency syndrome, and 1 of whom was immunocompetent. Both were treated with antibiotics, and neither developed systemic complications of the infection. These cases suggest that STEC infection should be considered in the differential diagnosis of chronic diarrhea. PMID- 15356802 TI - Influenza surveillance in Indonesia: 1999-2003. AB - Although influenza is recognized for its worldwide importance, little is known about the disease from tropical countries like Indonesia. From August 1999 through January 2003, a surveillance study was conducted in clinics at 6 sentinel locations. Adults (age, >14 years) and children (age, 4-14 years) presenting with respiratory symptoms suggestive of influenza were asked to enroll in the study. Nasal and pharyngeal swabs were examined by virus isolation, polymerase chain reaction, and rapid immunochromatographic tests. A total of 3079 specimens were collected from 1544 participants. Influenza infection was confirmed in 172 volunteers (11.1%) presenting with influenza-like illness. Influenza A (H1N1 and H3N2) and B viruses were detected at all sites. Peak prevalence tended to coincide with the respective rainy seasons, regardless of location. In light of the recent epidemic of severe acute respiratory syndrome, continued influenza surveillance would be useful in strengthening the infrastructure of the Indonesian public health system. PMID- 15356803 TI - Nontuberculous mycobacterial infection in children: a 2-year prospective surveillance study in the Netherlands. AB - We performed a prospective, 2-year nationwide study to assess incidence and disease characteristics of suspected infections with nontuberculous mycobacteria (NTM) in children, via the Netherlands Pediatric Surveillance Unit. Data for 61 children were reported (median age, 31 months; interquartile range, 22-50 months; female sex, 37 subjects); 2 subjects had an underlying disease. Most children (53 [87%] of 61) had cervical lymph node enlargement, with abscess in 25 (47%) and fistula in 11 (21%). The estimated annual incidence of NTM infection was 77 cases per 100,000 children. In 16 children, the diagnosis was based solely on the results of skin tests with mycobacterial antigens. Cultures were performed in 36 cases and yielded mycobacteria in 27 (75%); Mycobacterium avium was isolated from 18 cultures. Children with a culture positive for mycobacteria did not differ in presentation, complications, or treatment from those whose cultures showed no growth. Thirty children underwent surgery, and chemotherapy was the single treatment in 24 (39%) of the cases. The treatment of localized NTM infection in immunocompetent children by antimycobacterial drugs should be evaluated further. PMID- 15356804 TI - Nontuberculous mycobacteria: clinical importance and relevance to bacille Calmette-Guerin vaccination. PMID- 15356805 TI - Detection and control of influenza outbreaks in well-vaccinated nursing home populations. AB - BACKGROUND: Influenza outbreaks continue to occur in nursing homes despite high vaccination coverage among residents. Recommendations for outbreak control in institutions such as nursing homes advises use of antiviral drugs to reduce influenza transmission. METHODS: Influenza surveillance was performed among elderly residents of nursing homes in Michigan during 2 influenza seasons. The antiviral drug oseltamivir was used for outbreak control at the discretion of nursing home staff once influenza transmission was confirmed by virus isolation or rapid antigen detection. RESULTS: During 2000-2001, influenza was not confirmed in any of the 28 participating homes, despite transmission of types A (H1N1) and B in the community. During 2001-2002, influenza type A (H3N2) transmission was confirmed in 8 (26%) of 31 participating homes; influenza vaccine coverage among residents was 57%- 98% in outbreak-associated homes. Oseltamivir was used in all homes with influenza transmission; outbreak control varied according to the rapidity of outbreak recognition and the extent of antiviral use. Reported adverse events were primarily gastrointestinal reactions and rashes. Analysis of the usefulness of rapid antigen detection tests for outbreak recognition indicated a sensitivity of only 77% (specificity, 92%). CONCLUSIONS: Oseltamivir was reasonably well tolerated, and its use, along with continued promotion of vaccination coverage among nursing home residents and staff, should be a valuable addition to institutional outbreak-control strategies. PMID- 15356806 TI - The full-court press for influenza prevention in elderly persons. PMID- 15356807 TI - The role of diet in the treatment of travelers' diarrhea: a pilot study. AB - A pilot study was performed to compare the effects of a restricted physiologic diet in 48 subjects with those of an unrestricted diet in 57 subjects on the duration and symptoms of acute travelers' diarrhea among US adults being treated with an antimicrobial agent in Mexico. Restricted physiologic diet was defined as the avoidance of certain foods during diarrheal illness, as specified in limited published literature. The mean duration of diarrhea (37 vs. 33 h) and clinical symptoms were similar between those practicing the restricted diet and those practicing unrestricted diets. These results suggest that restricting diet during treatment of travelers' diarrhea with an antimicrobial agent is not associated with improvement of clinical symptoms or with decreased duration of diarrhea. However, a much higher number of subjects would need to be studied to prove this point statistically. PMID- 15356808 TI - Diet in the treatment of diarrhea: from tradition to evidence. PMID- 15356809 TI - Indicators of potentially drug-resistant bacteria in severe nursing home-acquired pneumonia. AB - BACKGROUND: Knowledge of the determinant factors responsible for the presence of antimicrobial-resistant pathogens in severe nursing home-acquired pneumonia (NHAP) is deemed essential for antibiotic selection. METHODS: Data for institutionalized patients with cases of severe pneumonia confirmed by culture of protected bronchoalveolar lavage fluid samples (> or =10(3) cfu/mL) during a 36 month period were analyzed. A classification tree with a sensitivity of 100% was developed using binary recursive partitioning to predict which patients are unlikely to have drug-resistant pathogen (DRP)-related pneumonia. RESULTS: Of the 88 patients who satisfied the inclusion criteria, 17 had at least 1 DRP recovered from the lower respiratory tract. The predictor variables were the Activity of Daily Living score and previous use of antibiotics. Prospective application of the model in 47 patients over a 24-month period yielded a sensitivity of 100% (95% confidence interval [CI], 71.3%-100%) and a specificity of 69.4% (95% CI, 51.9%-83.6%). CONCLUSIONS: The use of the tree may provide a more rational basis for selecting initial therapy for severe NHAP after it is validated in a large prospective study. PMID- 15356810 TI - Epidemiological typing of community-acquired methicillin-resistant Staphylococcus aureus isolates from children in Taiwan. AB - BACKGROUND: A 1400-bed tertiary medical center in northern Taiwan was used to conduct an epidemiological study of children hospitalized with community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infection during a 5-year period. METHODS: Nineteen previously healthy children with predominantly skin and soft-tissue CA-MRSA infections were enrolled into the study. Seventeen CA-MRSA isolates were examined for antimicrobial susceptibility and molecular typing. RESULTS: A comparison of our results with the reported resistance rates among CA MRSA isolates from other countries showed uniformly high macrolide resistance (100%). Of the 17 MRSA isolates in our study, all had the macrolide-lincosamide streptogramin-constitutive phenotype and the ermB gene. Moreover, on the basis of molecular typing results, 11 (65%) of 17 CA-MRSA isolates were genetically related (as determined by pulsed-field gel electrophoresis), and multilocus sequence typing revealed a sequence type of 59 in all isolates. Staphylococcal toxin genes lukS-PV and lukF-PV were detected in all isolates. However, staphylococcal cassette chromosome mec type IV was only detected in 3 (17.6%) of 17 isolates; the remaining 14 isolates were untypeable. CONCLUSIONS: Analysis of our data suggests the predominance of a single endemic CA-MRSA strain with high macrolide resistance in our community. Clinical improvement with incision and drainage was noted for most patients, despite treatment with an ineffective antibiotic, so the need for a change in treatment guidelines should be addressed. PMID- 15356811 TI - Hepatotoxicity of rifampin-pyrazinamide and isoniazid preventive therapy and tuberculosis treatment. AB - BACKGROUND: Severe liver injury has been attributed to preventive treatment of latent tuberculosis infection with a 2-month course of rifampin-pyrazinamide. METHODS: A retrospective cohort study in The Netherlands compared the hepatotoxicity of preventive treatment with rifampin-pyrazinamide with that of preventive treatment with isoniazid, and also with that of treatment for active tuberculosis containing at least isoniazid, rifampin, and pyrazinamide. RESULTS: Preventive treatment with rifampin-pyrazinamide caused severe hepatotoxicity more often than did preventive treatment with isoniazid (odds ratio [OR], 2.61; 95% confidence interval [CI], 1.26-5.39; P=.012), especially in patients <25 years old. It also caused severe hepatotoxicity more often than triple- or quadruple drug tuberculosis treatment (OR, 2.61; 95% CI, 1.21-5.59; P=.016), especially if the pyrazinamide dose was > or =30 mg/kg. Preventive treatment with rifampin pyrazinamide was more hepatotoxic even when the advised pyrazinamide dose of up to 20 mg/kg for preventive treatment was compared with the pyrazinamide dose of 30 mg/kg for tuberculosis treatment. CONCLUSIONS: Preventive treatment with rifampin-pyrazinamide causes severe hepatotoxicity more often than does preventive treatment with isoniazid or curative treatment for tuberculosis. PMID- 15356812 TI - Predicting hospital rates of fluoroquinolone-resistant Pseudomonas aeruginosa from fluoroquinolone use in US hospitals and their surrounding communities. AB - Rates of fluoroquinolone resistance among Pseudomonas aeruginosa in hospitals are increasing, but interhospital variability is great. We sought to determine whether this variability correlated to fluoroquinolone use in hospitals and in the surrounding community. Hospital quinolone use in 1999 (24 hospitals) through 2001 (35 hospitals) was determined from billing records. The number of fluoroquinolone prescriptions within a 10-mile (approximately 16-km) radius of each hospital was determined for 1999 and 2000. Hospital fluoroquinolone use increased from 1999 through 2001, from 137 to 163 defined daily doses (DDD)/1000 patient-days (P=.01). The rate of community fluoroquinolone use also increased, from 2.3 to 2.8 DDD/1000 inhabitant-days (P<.001). Rates of fluoroquinolone resistant P. aeruginosa increased from 29% in 1999 to 36% in 2001 (P=.003). Both community and hospital fluoroquinolone use were predictive of rates of fluoroquinolone-resistant P. aeruginosa. Levofloxacin was associated with resistance, but ciprofloxacin was not. Most of the variability in resistance rates is explained by volume of fluoroquinolone use, both in the hospital and the surrounding community. PMID- 15356813 TI - Health sequelae of human cryptosporidiosis in immunocompetent patients. AB - BACKGROUND: There have been no systematic studies following up the longer term health effects of cases of cryptosporidiosis for which genotype data exist. METHODS: We report a follow-up study of cases of laboratory-confirmed cryptosporidiosis. Case patients were sent a postal questionnaire asking about a wide range of symptoms occurring within 2 months after their initial diagnosis, and control subjects were sent the questionnaire 2 months after they had been recruited to the original study. RESULTS: Completed questionnaires were received from 235 case patients and 232 control subjects. For 111 of the case patients, the species of the infecting strain was known; 61 of these strains were Cryptosporidium hominis (human genotype), and 50 were Cryptosporidium parvum (bovine genotype). Forty percent of the case patients reported recurrence of intestinal symptoms after resolution of the acute stage of illness, irrespective of whether infection was with C. hominis or C. parvum. Reports of joint pain (odds ratio [OR], 2.8), eye pains (OR, 2.44), recurrent headache (OR, 2.10), dizzy spells (OR, 1.69), and fatigue (OR, 3.0) were significantly more common in case patients than in control subjects, but only in people who had experienced C. hominis infection. Joint symptoms experienced by case patients were of longer duration than those experienced by control subjects. CONCLUSIONS: Our results confirm previous reports of a high rate of relapse of gastrointestinal symptoms following recovery from an acute episode of cryptosporidiosis and show that C. hominis but not C. parvum is associated with an increased risk of nonintestinal sequelae. This study demonstrates that the impact of cryptosporidiosis on public health extends beyond that of the acute diarrheal illness and can lead to significant health sequelae. PMID- 15356814 TI - Increase in methicillin-resistant Staphylococcus aureus acquisition rate and change in pathogen pattern associated with an outbreak of severe acute respiratory syndrome. AB - BACKGROUND: An outbreak of severe acute respiratory syndrome (SARS) occurred in our 22-bed intensive care unit (ICU; Prince of Wales Hospital, Hong Kong, HKSAR, China) from 12 March to 31 May 2003, when only patients with SARS were admitted. This period was characterized by the upgrading of infection control precautions, which included the wearing of gloves and gowns all the time, an extensive use of steroids, and a change in antibiotic prescribing practices. The pattern of endemic pathogenic organisms, the rates of acquisition of methicillin-resistant Staphylococcus aureus (MRSA), and the rates of ventilator-associated pneumonia (VAP) were compared with those of the pre-SARS and post-SARS periods. METHODS: Data on pathogenic isolates were obtained from the microbiology department (Prince of Wales Hospital). Data on MRSA acquisition and VAP rates were collected prospectively. MRSA screening was performed for all ICU patients. A case of MRSA carriage was defined as an instance in which MRSA was recovered from any site in a patient, and cases were classified as imported or ICU-acquired if the first MRSA isolate was recovered within 72 h of ICU admission or after 72 h in the ICU, respectively. RESULTS: During the SARS period in the ICU, there was an increase in the rate of isolation of MRSA and Stenotrophomonas and Candida species but a disappearance of Pseudomonas and Klebsiella species. The MRSA acquisition rate was also increased: it was 3.53% (3.53 cases per 100 admissions) during the pre SARS period, 25.30% during the SARS period, and 2.21% during the post-SARS period (P<.001). The VAP rate was high, at 36.5 episodes per 1000 ventilator-days, and 47% of episodes were caused by MRSA. CONCLUSIONS: A SARS outbreak in the ICU led to changes in the pathogen pattern and the MRSA acquisition rate. The data suggest that MRSA cross-transmission may be increased if gloves and gowns are worn all the time. PMID- 15356815 TI - Infected bilomas in liver transplant recipients: clinical features, optimal management, and risk factors for mortality. AB - BACKGROUND: Infected hepatic fluid collections (bilomas) are a major infectious complication of liver transplantation. Limited data exist on management and outcome of biloma. METHODS: We report a cohort study of 57 liver transplant recipients with posttransplantation bilomas undertaken to identify the clinical features of biloma, management strategies, and outcome. RESULTS: Fever (44%) and abdominal pain (40%) were the most common presenting symptoms, but one-third of patients were asymptomatic; 79% had elevated hepatic enzyme levels. Patients without hepatic artery thrombosis (HAT) had the highest rates of resolution with percutaneous drainage and anti-infective therapy (64%). Retransplantation was necessary in 64% of patients with HAT and biloma. Independent predictors of resolution with nonsurgical therapy were absence of HAT (odds ratio [OR] 7.69; P=.01) and absence of Candida (OR, 9.09; P=.02) or enterococcal infection (OR, 7.69; P=.03). Patients with bilomas had significantly greater mortality (Cox proportional hazard ratio [HR], 2.38; P=.008, by log rank test) and graft loss (HR, 4.31; P<.0001). Predictors of mortality by multivariable analysis included renal insufficiency (OR, 12.51; P=.02) or infection with Candida species (OR, 4.93; P=.03) or gram-negative bacilli (OR, 9.12; P=.01). CONCLUSION: Posttransplantation biloma should be suspected in patients with fever or abdominal pain or abnormalities of hepatic enzymes, and it can be confirmed by computerized tomography and radiographically guided aspiration. Bilomas are most likely to be successfully treated nonsurgically in patients without HAT and without Candida or enterococcus infection. PMID- 15356816 TI - Challenges in the design of antibiotic equivalency studies: the multicenter equivalency study of oral amoxicillin versus injectable penicillin in children aged 3-59 months with severe pneumonia. AB - The World Health Organization (WHO) recommends that children with severe pneumonia (characterized by cough or difficult breathing, as well as lower chest wall indrawing) be hospitalized and treated with parenteral penicillin. Oral amoxicillin, if equally effective for treating severe pneumonia, would address challenges associated with providing parenteral therapy, including risk of transmission of bloodborne pathogens from contaminated needles, exposure to nosocomial pathogens during hospitalization, inadequate access to health care facilities, and cost. The recently completed multicenter international trial of oral amoxicillin versus parenteral penicillin for treatment of severe pneumonia demonstrated the equivalency of these agents in children with severe pneumonia. This article focuses on the challenges of designing an equivalence study and the threats to the validity of the trial results, particularly the implications of the bias toward finding equivalence when subjects are unlikely to respond to either study therapy. These considerations have implications for use of the Amoxicillin Penicillin Pneumonia International Study (APPIS) results in clinical practice and for potential modification of WHO treatment guidelines. PMID- 15356817 TI - Sexually transmitted infections in travelers: implications for prevention and control. AB - Sexually transmissible diseases (STDs), the most common notifiable infectious conditions, remain major threats to reproductive and public health worldwide. Travelers are particularly vulnerable to STDs, because of voluntary or involuntary sexual behavior while abroad, and are significant vectors who introduce new pathogens and resistant strains to unaffected parts of the world. This article outlines some key issues that travel medicine specialists and other clinicians should revisit when providing services to travelers. We discuss obstacles to promoting sexual health, including the diversity of the target group, unanticipated opportunities for sexual risk, ambivalent cooperation by the travel and tourism industries, poorly developed travel health sectors, illegal migration and sex tourism, and lack of research about the association between travel and STDs. We also outlined some programmatic aspects of public health that should be identified and addressed for the promotion of sexual health among travelers. PMID- 15356818 TI - Staphylococcus aureus with reduced susceptibility to vancomycin. AB - Infections with Staphylococcus aureus with reduced susceptibility to vancomycin continue to be reported, including 2 cases caused by S. aureus isolates with full resistance to vancomycin. This review first outlines the definitions of vancomycin-intermediate S. aureus (VISA) and vancomycin-resistant S. aureus (VRSA) and risk factors for infection. Next, we describe the mechanisms of resistance and methods of laboratory detection of the organisms. Finally, we address infection control and management issues associated with isolation of VISA and VRSA. PMID- 15356819 TI - Antimicrobial resistance in nontyphoid Salmonella serotypes: a global challenge. AB - Increasing antimicrobial resistance in nontyphoid Salmonella species has been a serious problem for public health worldwide. The high rate of resistance is hampering the use of conventional antibiotics, and growing resistance to newer antimicrobial agents is aggravating the situation. The circumstances of occurrence and spread of antimicrobial resistance are complex; however, a major cause is the widespread use of antimicrobial agents in food animals, particularly in animal feed. Genetic analysis has indicated that the source of resistance is frequently a transferable plasmid. Recent studies have revealed that some serotype-specific virulence plasmids form hybrid plasmids through recombination with resistance plasmids or acquire gene cassettes consisting of multiple resistance genes. Such evolutionary events provide a virulent strain the advantage of survival in an unfavorable drug environment. In view of the serious implications associated with drug-resistant Salmonella species, a more deliberate use of antibiotics in both human medicine and animal industry is warranted. Continued surveillance of antimicrobial resistance and use of antimicrobial agents in food animals is also indispensable. PMID- 15356820 TI - HIV-1 drug resistance in subjects with advanced HIV-1 infection in whom antiretroviral combination therapy is failing: a substudy of AIDS Clinical Trials Group Protocol 388. AB - We evaluated phenotypic and genotypic markers of drug resistance in human immunodeficiency virus type 1 (HIV-1) at the time of virologic failure (VF) in subjects in the AIDS Clinical Trials Group Protocol 388 (ACTG 388) who received lamivudine-zidovudine (or lamivudine-stavudine) and either indinavir, efavirenz indinavir, or nelfinavir-indinavir. At VF, phenotypically susceptible HIV-1 was found in 55% of subjects in the nelfinavir-indinavir arm, compared with 22% in the indinavir arm (P=.006). Phenotypic resistance to lamivudine was less common in the efavirenz-indinavir arm (33% of subjects; P=.002) and the nelfinavir indinavir arm (43%; P=.003), compared with the indinavir arm (78%). Isolated phenotypic resistance to efavirenz at VF occurred in HIV-1 recovered from 33% of subjects in the efavirenz-indinavir arm; 24% of the subjects had HIV-1 with both efavirenz and lamivudine resistance. Results of genotypic tests were similar. The lower frequency of resistance in the nelfinavir-indinavir arm likely reflects decreased drug exposure that is due to intolerance, which is consistent with the lower potency and tolerability of this combination in ACTG 388. The lower frequency of lamivudine resistance in the efavirenz-indinavir arm is consistent with reports in other studies of potent regimens. Thus, although dual resistance to efavirenz and lamivudine occurred at VF in the efavirenz-indinavir arm, this risk was relatively low when evaluated in the context of the potency and tolerability of this regimen. PMID- 15356821 TI - When more is less. PMID- 15356823 TI - Rifampin and pyrazinamide for latent tuberculosis infection: clinical trials and general practice. PMID- 15356822 TI - Hepatotoxicity of rifampin and pyrazinamide in the treatment of latent tuberculosis infection in HIV-infected persons: is it different than in HIV uninfected persons? AB - BACKGROUND: In 2000, results of a multinational trial demonstrated that a 2-month course of rifampin and pyrazinamide (RZ) was as effective as isoniazid (INH) in reducing tuberculosis in human immunodeficiency virus (HIV)-infected individuals with latent tuberculosis infection (LTBI). After the release of new guidelines, the Centers for Disease Control and Prevention received reports of severe hepatotoxicity associated with the use of the RZ regimen for the treatment of LTBI in the general population. To better understand the occurrence of hepatotoxicity in an HIV-infected population, we conducted a more detailed analysis of the liver function test results obtained in the multinational trial of RZ. METHODS: At study entry, patients were required to have a bilirubin level of < or =2.5 mg/dL and both an aspartate aminotransferase (AST) level and an alkaline phosphatase level of < or =5 times the upper limit of normal. Patients with acute hepatitis were excluded. At months 1 and 2 of the study, all patients had bilirubin and AST levels measured. RESULTS: There was no difference between the RZ and INH groups with regard to AST level or bilirubin level at baseline. An increase in the AST level of > or =40 U/L was associated with the use of INH and older age; and an increase in the bilirubin level of > or =0.5 mg/dL was associated with the use of RZ, male sex, and nonwhite race (P<.05). An absolute AST level of >250 U/L occurred in 12 of 745 INH recipients and in 15 of 721 RZ recipients (P=.56), and an absolute bilirubin level of >2.5 mg/dL occurred in 5 of 743 INH recipients and 13 of 718 RZ recipients (P=.06). CONCLUSIONS: These data demonstrate very little liver injury associated with either INH or RZ in the HIV-infected subjects, leaving unclear the reasons for serious RZ-related liver damage in the general population. PMID- 15356824 TI - Long-term clinical follow-up, without antiretroviral therapy, of patients with chronic HIV-1 infection with good virological response to structured treatment interruption. AB - The present study assessed the long-term clinical follow-up of 26 of 60 human immunodeficiency virus (HIV)-positive patients who followed structured treatment interruption (STI) protocols and who, because of good virological response, did not resume receipt of highly active antiretroviral therapy (HAART). The plasma viral load (pVL) noted after > or =2 years without antiretroviral therapy remained significantly lower than the pVL noted before initiation of HAART, for 11 of the 26 patients (i.e., for 18% of the 60 patients who had STI performed). The CD4+ T cell count remained stable throughout the study. A low pVL at the end of follow-up was independently associated with a low CD8+CD38+ T lymphocyte count and a high stimulation index to p24 antigen after STI. In conclusion, approximately 2 years after STI, only a low proportion of patients who had a good immunological profile at the end of the STI period still had a good virological response. PMID- 15356825 TI - K103N mutation in antiretroviral therapy-naive African patients infected with HIV type 1. AB - Most of the information about genetic sequencing and drug susceptibility of human immunodeficiency virus type 1 (HIV-1) is derived from the study of HIV-1 subtype B. Worldwide, most people infected with HIV-1 are infected with non-subtype B viruses and live in developing countries. We report 3 cases of antiretroviral naive African immigrants infected with HIV-1 strains possessing the K103N mutation in the reverse transcriptase gene, which confers high-level resistance to all nonnucleoside reverse transcriptase inhibitors. PMID- 15356826 TI - The search for a predictor of CD4 cell count continues: total lymphocyte count is not a substitute for CD4 cell count in the management of HIV-infected individuals in a resource-limited setting. AB - Total lymphocyte count (TLC) has been recommended as a substitute for CD4 cell count for the management of HIV-infected individuals living in resource-limited settings. To confirm this, 151 TLCs and CD4 cell counts were obtained from 109 patients who had not yet started treatment and analyzed. CD4 cell counts of <200 cells/mm3 were found in 42 cases (37.8%) with TLCs of > or =1200 cells/mm3. Thus, 1 in 3 individuals would have been deprived of needed treatment. Therefore, in this setting, TLC is not a reliable predictor of CD4 cell count in HIV-infected individuals. PMID- 15356827 TI - Invasive zygomycosis in hematopoietic stem cell transplant recipients receiving voriconazole prophylaxis. AB - We report 4 cases of invasive zygomycosis in hematopoietic stem cell transplant recipients, all occurring after May 2003, when voriconazole began to be used as antifungal prophylaxis. No cases of zygomycosis had been detected in this population in the 3 years prior to May 2003. All 4 patients were receiving immunosuppressive therapy for presumed graft-versus-host disease. Profoundly immunosuppressed patients receiving voriconazole prophylaxis remain at risk for less-common pathogens that are intrinsically resistant to this agent. PMID- 15356828 TI - Zygomycosis: reemergence of an old pathogen. PMID- 15356829 TI - A 42-week open-label study to assess the pharmacokinetics, antiretroviral activity, and safety of amprenavir or amprenavir plus ritonavir in combination with abacavir and lamivudine for treatment of HIV-infected patients. AB - The pharmacokinetics, antiviral activity, and safety of an amprenavir-ritonavir (APV-RTV) 600/100 mg b.i.d. regimen and an APV-RTV 1200/200 mg q.d. regimen were studied in a human immunodeficiency virus (HIV)-infected population. The geometric least-square mean ratio (90% confidence interval) of steady-state trough concentrations, compared with that of the amprenavir 1200 mg b.i.d. regimen, was 6.08 (4.94-7.49) for the twice-daily APV-RTV regimen, and it was 4.19 (2.90-6.08) for the daily APV-RTV regimen. The regimens were well tolerated, which supports APV-RTV as an option for twice-daily or daily therapy for HIV. PMID- 15356830 TI - A summer outbreak of influenza A virus infection among young children. AB - In the summer of 2003 in Jerusalem, Israel, 23 children were hospitalized with influenza A virus (A/Fujian/411/02-like virus) infection. The majority were neonates and infants. Clinical manifestations included neonatal fever, bronchitis, bronchiolitis, and pneumonia, and outcomes were favorable. Continued surveillance between epidemic seasons could allow early recognition of influenza strains that will appear in the following influenza season. PMID- 15356831 TI - Can we describe the epidemiology of candidemia without using selective blood culture bottles for fungus detection? PMID- 15356833 TI - Risk factors for extrapulmonary tuberculosis. PMID- 15356835 TI - Effectiveness of environmental and infection control programs to reduce transmission of Clostridium difficile. PMID- 15356836 TI - The incidence of Clostridium difficile-associated and non-C. difficile-associated diarrhea after use of gatifloxacin and levofloxacin in an acute-care facility. PMID- 15356837 TI - Pharmacokinetics of voriconazole in the cerebrospinal fluid of an immunocompromised patient with a brain abscess due to Aspergillus fumigatus. PMID- 15356838 TI - Use of clinical criteria and molecular diagnosis to more effectively monitor patients recovering after severe acute respiratory syndrome coronavirus infection. PMID- 15356839 TI - Concurrent cerebral american trypanosomiasis and toxoplasmosis in a patient with AIDS. AB - We report a case of concurrent cerebral infection with Trypanosoma cruzi and Toxoplasma gondii in a patient with acquired immunodeficiency syndrome (AIDS). A 22-year-old El Salvadoran man initially improved during receipt of antitoxoplasmosis therapy, but he had rapidly progressive hemiplegia. Magnetic resonance imaging showed an abnormal finding in the left internal capsule, and cytological analysis of cerebrospinal fluid revealed T. cruzi trypomastigotes. Despite prompt therapy with nifurtimox, the patient's mental status declined, and he died of nosocomial complications. Although infrequent, T. cruzi infection should be considered in the differential diagnosis of brain lesions in patients with AIDS from regions of endemicity. PMID- 15356840 TI - [GIN changes typographical format and objectives]. PMID- 15356841 TI - [New concepts of tubular calcium transport in the kidney: clinical implications]. AB - The kidney plays an important role in calcium homeostasis. In this review we discuss new concepts in tubular calcium transport, related proteins and the clinical implications of these new findings. Most calcium reabsorption occurs in the proximal tubules via the passive paracellular pathway, but calcium reabsorption also occurs in the thick ascending limb of Henle's loop (50% via the transcellular pathway). Finally, at the level of the distal convoluted tubule and connecting tubule calcium is reabsorbed via the active transcellular route. The Calcium-sensing receptor, localised along the thick ascending limb of Henle's loop, regulates the urinary calcium excretion in response to changes in extracellular calcium concentration. The Epithelial Calcium Channel 1 is a highly Ca2+-sensitive channel that is predominantly present in the connecting tubule. The Calbindin D(28k) is a cytoplasmatic protein expressed in the distal tubule, it is involved both in transcellular calcium diffusion and in the control of intracellular calcium concentration. Heterozygous mutations in the gene for the calcium sensing receptor, which result in a loss of function by the receptor, are associated with familial hypocalciuric hypercalcaemia. Mutations involving a gain of function have been associated with hypocalcaemia with normal serum parathyroid hormone concentration. Bartter's syndrome, caused by a dysfunction of thick ascending limb cells, is associated with calcium wasting. On the contrary, Gitelman's syndrome, caused by a dysfunction of the distal tubule, is characterised by hypocalciuria and hypomagnesemia. PMID- 15356842 TI - [Psychosocial adaptation in adolescents with chronic kidney disease]. AB - BACKGROUND: Chronic disease represents a very critical experience for personal identity. This is far more true in adolescence when corporeal, emotional, relational and cognitive changes are taking place. The aim of the present study is to draw attention to the social and emotional balance and the psychosocial adaptation in a group of nephropathic adolescents and to formulate a suitable assistance program. METHODS: We studied two adolescent groups, one group of nephropathics and one group of healthy individuals, by administering the MMPI (Minnesota Multiphasic Personality Inventory) personality test. RESULTS: We found significant differences between the mean score of the two groups in the clinical scale for depression, hypochondria and hysteria (p < 0.5); in addition, we observed a statistically significant difference in the emotional balance indicators. CONCLUSIONS: Our experience suggests that nephropathic adolescents require global awareness of their psychological problems through the use of innovative assistance methods. A new strategy involving a more modern approach (i.e., educational camps) and specific psychosocial rehabilitation programs should be introduced to improve the quality of life. PMID- 15356843 TI - [Natural history of HBV in dialysis population]. AB - Dialysis patients remain at risk of acquiring hepatitis B virus (HBV) infection. The issue of the natural history of HBV among patients undergoing long-term dialysis remains unclear. Assessing the natural history of hepatitis B in patients on maintenance dialysis is problematic because of the unique characteristics of this population: serum aminotransferase activity is lower in dialysis patients compared with patients without renal disease; also, chronic hepatitis B has an insidious and prolonged natural history, and the competing mortality from complications of end-stage renal disease may obscure the long-term consequences of hepatitis B. HBV-related liver disease frequently runs an asymptomatic course in dialysis patients and the liver-related mortality in this population is very low; thus, the prognosis for chronic HBV infection in dialysis patients has been reported as benign. However, the frequency of liver cancer in dialysis patients appears higher than that observed in the general population, this has been related to a greater exposure to HBV/HCV. Cirrhosis is not a frequent comorbid condition in the dialysis population of industrialised countries, but the death rate for dialysis patients with cirrhosis is 35% higher than for those without it. In addition, it has been observed that liver disease remains a significant cause of mortality among HbsAg-positive carriers on dialysis in developing countries. The low viral load measured in dialysis patients with persistent HBsAg carriage could be accounted for by the relatively benign course of HBV-related liver disease in this population. Prospective clinical trials are under way to better define the virological features of HBV in the dialysis population. PMID- 15356844 TI - [Respiratory function abnormalities in uremic patients]. AB - Patients with end-stage renal disease treated by hemodialysis are exposed to continuous pulmonary insults of multifactorial origin. Alterations in respiratory drive, mechanics, muscle function and gas exchange are frequent in hemodialysis patients. Pulmonary dysfunction may be the direct consequence of circulating uraemic toxins or may result indirectly from volume overload, anaemia, immune suppression, extraosseous calcification, malnutrition, electrolyte disorders, and/or acid-base imbalances. We have emphasised how derangement of diffusing capacity represents the most frequent and important respiratory abnormality in haemodialysed patients. It has been postulated that some forms of selective damage in the alveolo-capillary wall interferes with alveolar gas exchange. PMID- 15356845 TI - [Changes is Th1/Th2 balance in hemodialysis patients: effect of different membranes]. AB - BACKGROUND: Hemodialysis (HD) patients present an immunodeficiency that is mainly related to the defect of cell-mediated immunity. We have previously shown the polarisation of T-helper cells toward the phenotype in HD treatment with cuprophan membrane. In the present study, we have examined the effect of a Vitamin E-coated dialyser (Excebrane, VE) on the activity of the two Th subsets. METHODS: We studied 8 healthy controls and 10 patients on RDT for at least 6 months with cellulose membrane (AC), then switched to HD-VE. Blood was collected from HD patients while on treatment with AC, and after 1 year of treatment with VE. CD4+ cells were isolated from peripheral blood mononuclear cells (PBMC) by negative selection, treating PBMC with a cocktail of anti-CD8, -CD16, -CD19, CD36 and -CD56 antibodies labelled with magnetic beads, and passing them through a magnetic field. The collected Th cells were cultured for 48 h with and without phytohemagglutinin (PHA). IFNgamma and IL-4 were measured in the supernatant using the ELISA assay. RESULTS: The constitutive release of IL-4 by CD4+ cells was abated significantly by treatment with VE. IFNgamma released by mitogen stimulated CD4+ recovered with VE. CONCLUSIONS: This study demonstrates that treatment with vitamin E-coated dialyser improves the defect of PBMC function associated with cellulose membrane dialyser consisting of altered spontaneous and mitogen-stimulated cytokine release. The effects of vitamin E-coated filter, in particular the recovery of reactive IFNgamma production by Th1 cells and the restriction of spontaneous IL-4 release by Th2 cells may have clinical importance. PMID- 15356846 TI - [Diabetes and atherosclerotic coronary damage in uremia]. AB - BACKGROUND: Cardiovascular risk factors are common findings in uraemics, but the impact of each single factor on the development of atherosclerosis is still a matter of debate. PATIENTS AND METHODS: In order to evaluate the relationship between diabetes and ischaemic heart disease (IHD) in uraemia, we carried out a retrospective study comparing the results of 33 coronary angiographies performed in non-diabetic patients with those of 13 diabetics (2 had type 1 diabetes, 8 were treated with insulin, 2 with sulfonylureas and 3 received no therapy). Coronary angiography was performed in 29 patients awaiting kidney transplantation and in 17 subjects with IHD. RESULTS: Age, sex, length of time on renal replacement therapy, smoking history, clinical diagnosis of cerebrovascular and peripheral vascular disease, systolic blood pressure (BP), cholesterol, triglycerides, calcium, phosphate, albumin and degree of anaemia were comparable in the two groups. On the contrary, frequency of IHD (77 vs. 30%, p<0.01) and atrial fibrillation (23 vs. 3%, p<0.05) were higher, while diastolic BP (79 +/- 7 vs. 85 +/- 8 mmHg, p<0.05) and calcium phosphate product (47 +/- 10 vs. 57 +/- 15 mg2/dL2, p<0.05) were lower in diabetics than in non-diabetics. Stenotic lesions of the three major coronary arteries were more prevalent in diabetics than in non diabetics (left anterior descending artery (LAD) 100 vs. 48%, p<0.01; right coronary artery (RCA) 77 vs. 39%, p<0.05; left circumflex artery (LCA) 69 vs. 24%, p<0.01) and in the same way diabetics showed higher narrowing percentage (LAD 74 +/- 30 vs. 30 +/- 36%, p<0.01; RCA 71 +/- 41 vs. 26 +/- 38, p<0.01; LCA 41 +/- 38 vs. 15 +/- 29, p<0.05). CONCLUSIONS: Our study demonstrates that although the uraemic milieu is a risk factor for IHD, diabetes increases the degree of atherosclerotic vascular damage independently of the other cardiovascular risk factors. PMID- 15356847 TI - [Impact of left ventricular patterns and diastolic dysfunction on hemodialysis patients]. AB - BACKGROUND: Left ventricular hypertrophy is the main manifestation of uraemic cardiomyopathy and predicts both cardiovascular events and death independently of conventional risk factors. Diastolic dysfunction is often associated with left ventricular hypertrophy, and accounts for up to 30% of heart failure. The aim of this study was to estimate the impact and the prevalence of left ventricular hypertrophy, its geometric models and diastolic dysfunction on haemodialysis patients, as well as the relationship with cardiac troponin, a specific marker for myocardial damage. METHODS: We enrolled in the study 31 patients (26 on haemodialysis, 5 on peritoneal dialysis) and 31 normal healthy subjects as the control group. Echocardiographic measurements were carried out according to the recommendations of the American Society of Echocardiography. Left ventricular mass was calculated according to the Devereux formula and indexed to height 2.7 We carried out Doppler echocardiography to study the diastolic function by measurements of isovolumic relaxation period (IVRT), E-wave deceleration time (DTE) and E/A ratio. We measured cardiac troponin using a third generation electrochemiluminescence immunoassay. We did the statistical analysis with the t test for comparison among groups, and we established correlations using the Pearson and Spearman correlation test. We considered values of p<0.05 to be statistically significant. RESULTS: Eccentric hypertrophy was the most frequent pattern (n=17; 55%), followed by normal cardiac geometry (n=7; 23%) and concentric hypertrophy (n=5; 16%). Only a minority of patients (n=2; 6%) showed concentric remodelling. Systolic dysfunction was present in 3 patients (EF<50%). Diastolic dysfunction was present in 24 out of 31 HD patients, that had abnormal relaxation pattern, characterised by prolonged IVRT, prolonged DTE and E/A ratio <1. Only one patient had restrictive filling pattern with reduced DTE and E/A ratio >2. Fourteen patients (45%) had E/A ratio < or =0.5, 10 patients (32%) had E/A ratio >0.5<1 and 7 patients (13%) had E/A ratio > or =1. DTE was prolonged in 14 patients (45%) and IVRT was prolonged in 6 patients (19%). The patients with E/A ratio < or =0.5 (n=14; 45%) suffered major hypotensive episodes during dialytic sessions compared to patients with E/A ratio >0.5, we found this difference to be statistically significant (p < 0.01). We observed the E/A ratio to be negatively correlated with age (r = -0.41; p=0.02) whereas DTE was positively correlated with posterior wall thickness (r =0.36; p= 0.05) and interventricular septum thickness (r =0.45; p=0.01). Cardiac troponin was positively correlated with age (r =0.50; p=0.00), left ventricular mass (r =0.41; p=0.02), posterior wall thickness (r =0.41; p=0.02) and interventricular septum thickness (r =0.39; p=0.03) but not with diastolic dysfunction parameters. Serum cardiac troponin was elevated (> or =0.10 micro g/L) in 12 patients (38.7%) and was associated with eccentric hypertrophy in all the cases. We detected no significant differences among groups with normal left ventricular geometry and left ventricular hypertrophy in the duration of the dialytic treatment, but we noted a statistically significant difference in relationship to age (p=0.03). CONCLUSIONS: Left ventricular hypertrophy is associated with diastolic dysfunction, both cause of hypotensive episodes during dialytic treatments and heart failure in patients with normal systolic function. The non invasive assessment of left ventricular diastolic function would represent an important advancement in the diagnosis and prevention of heart failure in haemodialysis patients. PMID- 15356848 TI - [Non compliance predictive factors in renal transplantation]. AB - Currently many authors report a kidney graft survival of 90-95% at one years post transplantation. The patients' adherence to the immunosuppressive therapy is an important condition for maintaining graft functioning. A review of the medical literature shows that graft survival is significantly worse in non-compliance patients. As a matter of fact, 267 non compliance patients lost their graft of 7206 renal graft recipients. There are usually three different non-compliance profiles in transplantation : 1) 'Accidental non compliers' (47%) identifies those patients who sometimes forget to take the therapy. 2) 'Invulnerables' (28%) are those patients who believe that they do not need to take their immunosuppressive drugs regularly; 3) 'Decisive non compliers' (25%) are those patients who decide what therapy they should take. Many predictive factors have been associated with non compliance: 1) Demographic variables 2) Psychological variables 3) Psychiatric disorders 4) Poor social support 5) Pretransplantation compliance 6) Substance abuse We therefore think that it is particularly useful to identify non compliers before transplantation through psychological screening, in order to increase their compliance by supportive psychological assistance. Unfortunately, these patients tend to avoid specialists' intervention so it is very useful to provide the medical teams with the necessary training to cope with the psychological problems of their patients. PMID- 15356849 TI - [Management human resources]. AB - The management of human resources may follow different models, defined as bureaucratic, technocratic or managerial-entrepreneurial models. The latter being the most used. However, the relationship individual-enterprise is based on both a legal and a psychological contract regardless of the model used. The winning concept considers the personnel as the first and most important customer to be trained, informed and kept updated. For these reasons it is necessary to create a warm working environment, which is the first marketing tool, thus improving the marketing skills (enterprise-customer). The improved results (products, processes and publications) will be achieved by total quality management, which includes training and transformation of the chief's role from the hierarchical management to a coaching approach. This approach will recreativity, personality and competence of the personnel. This new type of leadership is based on the authority recognised by the personnel, service and motivation. PMID- 15356850 TI - [Search strategies for nephrology knowledge and evidence]. AB - The informatic techniques for searching biomedical literature have been one of the fundamental advantages of the era of information explosion. In this review, we focus on some methodologies for searching biomedical literature with particular focus on nephrology and describe the use of PubMed and the techniques for understanding and using the Medical Subject Heading (MeSH) to search Medline and other databases. Search strategies take advantages of simple search engines which generate informatic algorithms to find relevant articles; alternatively, a set of rules is to be known and represents a more powerful means for identification of selected publications. The latter technique is more useful for researchers seeking relevant information in the literature to prepare systematic reviews. Informatic techniques for searching biomedical literature have represented a fundamental advantage in science. Additional advantage will derive from the use of structured formats in research reporting, with better and quicker readability of retrieved published information, and from the effort of researchers in synthetizing the available published literature. PMID- 15356851 TI - [Tubolopaties associated to hypokalemia]. AB - We have described two clinical cases of two young men affected by hypkalemia associated with metabolic alkalosis. The first patient also presented hypercalciucia, normal magnesemia, defect in renal concentrating ability and increased renin activity; in addition he was affected by congenital sensorineural deafness. The diagnosis of Bartter's syndrome was made and it was confirmed by the gene analysis, which revealed a mutation for the beta-subunit of the ClC chloride channels known as barttin. The second case was characterized by hypocalciuria and hypomagnesemia, polyuria and nicturia. The genetic analysis revealed a mutation for the gene encoding the Na+-Cl(-) cotransporter and the diagnosis of Gitelman's syndrome was formulated. We present experimental and clinical evidence to explain, at the molecular level, the differences in calcium and magnesium homeostatis in the two cases. Moreover, we propose different causes to justify the pathogenesis of hypokalemia and the related metabolic alkalosis. PMID- 15356852 TI - [Acute renal failure from ACE inhibitors in elderly patients: are there any prognostic indicators?]. PMID- 15356853 TI - [Which nephrologist?]. PMID- 15356854 TI - Adolescent impulsivity predicts adult dominance attainment in male vervet monkeys. AB - Adolescence is characterized by behavioral and physiological changes that prepare individuals for the transition to adulthood. The purpose of this study was to evaluate the effects of behavioral, morphological, neurobiological, and developmental characteristics of adolescent male vervets in predicting later dominance attainment. Thirty-six adolescent male vervets were tested for social impulsivity by means of the Intruder Challenge test while they were still living in their natal groups. Body weight and cerebrospinal fluid (CSF) metabolites of serotonin, dopamine, and norepinephrine were measured before they were introduced into new matrilineal breeding groups at age 5. Stable adult dominance rank was determined at age 6, 1 year following introduction. The results indicated that body weight, adolescent impulsivity, and levels of 5-hydroxyindoleacetic acid (5 HIAA) and homovanillic acid (HVA) in CSF predicted adult dominance attainment. As expected, males that were above average in body weight prior to introduction were significantly more likely to become dominant. Males that were high in impulsivity as adolescents, and low in 5-HIAA prior to introduction were more likely to achieve stable alpha male status 1 year following introduction. The combination of these three factors resulted in correct prediction of rank attainment for 92% (33/36) of the males. Two other factors-maternal dominance rank and a measure of social anxiety from the Intruder Challenge test-were not related to adult dominance attainment in this sample. These results support the idea that there are benefits of a high-risk, high-gain strategy is beneficial for adolescent and young adult male vervets. They also demonstrate that adolescent impulsivity is age-limited. Males that achieved high rank moderated their behavior as adults, and no longer scored high in impulsivity relative to their age peers. PMID- 15356855 TI - Cross-species amplification of human microsatellite markers using noninvasive samples from white-handed gibbons (Hylobates lar). AB - Analysis of the population genetic structure and reproductive strategies of various primate species has been facilitated by cross-species amplification (i.e., the use of microsatellite markers developed in one species for analysis of another). In this study we screened 47 human-derived markers to assess their utility in the white-handed gibbon (Hylobates lar). Only eight produced accurate, reliable results, and exhibited levels of polymorphism that were adequate for individual identification. This low success rate was surprising given that human microsatellite markers typically work well in species (such as macaques) that are evolutionarily more distant from humans than are gibbons. In addition, we experienced limited success in using a set of microsatellite markers that have been reported to be useful in the closely-related H. muelleri, and applying our set of microsatellite markers to samples obtained from one H. pileatus individual. Our results emphasize the importance of extensively screening potential markers in representatives of the population of interest. PMID- 15356856 TI - Influence of the mother's reproductive state on the hormonal status of daughters in marmosets (Callithrix kuhlii). AB - Behavioral and endocrine suppression of reproduction in subordinate females produces the high reproductive skew that characterizes callitrichid primate mating systems. Snowdon et al. [American Journal of Primatology 31:11-21, 1993] reported that the eldest daughters in tamarin families exhibit further endocrinological suppression immediately following the birth of siblings, and suggested that dominant females exert greater control over subordinate endocrinology during this energetically challenging phase of reproduction. We monitored the endocrine status of five Wied's black tufted-ear marmoset daughters before and after their mother delivered infants by measuring concentrations of urinary estradiol (E(2)), pregnanediol glucuronide (PdG), testosterone (T), and cortisol (CORT). Samples were collected from marmoset daughters 4 weeks prior to and 9 weeks following three consecutive sibling-litter births when the daughters were prepubertal (M=6.1 months of age), peripubertal (M=11.9 months), and postpubertal (M=17.6 months). The birth of infants was associated with reduced ovarian steroid excretion only in the prepubertal daughters. In contrast, ovarian steroid levels tended to increase in the postpubertal daughters. Urinary E(2) and T levels in the postpubertal daughters were 73.8% and 37.6% higher, respectively, in the 3 weeks following the birth of infants, relative to prepartum levels. In addition, peak urinary PdG concentrations in peri- and postpubertal daughters were equivalent to luteal phase concentrations in nonpregnant, breeding adult females, and all of the peri- and postpubertal daughters showed clear ovulatory cycles. Cortisol excretion did not change in response to the reproductive status of the mother, nor did the concentrations change across age. Our data suggest that marmoset daughters of potential breeding age are not hormonally suppressed during the mother's peripartum period or her return to fertility. These findings provide an additional example of species diversity in the social regulation of reproduction in callitrichid primates. PMID- 15356857 TI - Proximate mechanisms of reproductive monopolization in male moustached tamarins (Saguinus mystax). AB - In moustached tamarin (Saguinus mystax) groups, the single breeding female mates polyandrously with most or all nonrelated adult males. Nonetheless, paternity is monopolized in many groups by a single male. No evidence for male endocrine suppression has been found in this species. The proximate mechanisms of monopolization thus remain poorly understood. The aim of this study was to investigate the possible impact of agonistic interactions and mate-guarding on the monopolization of paternity in male moustached tamarins. Furthermore, we evaluated the likely costs of these behaviors, and whether olfactory cues might be used for its timing. We used behavioral data on proximity, agonistic interactions, time budgets, and scent-marking behavior to answer these questions. While direct agonistic competition does not play a prominent role, fertile females were consorted in some periods by one male, the sire of the previous and next litter. Consorting was instigated nearly exclusively by the male. It probably occurred during the female's periods of highest fertility, and thus likely functions as mate-guarding. The timing of the consortship was probably guided by olfactory cues in the female's scent marks. While we did not obtain direct evidence for energy costs in terms of increased energy expenditure or decreased food intake, we found that consorting males are more conspicuous and therefore may be more vulnerable to predators. PMID- 15356858 TI - Sexual communication between breeding male and female cotton-top tamarins (Saguinus oedipus), and its relationship to infant care. AB - Male cotton-top tamarins have been shown to be responsive to female scent cues of ovulation, and are known to actively participate in infant care during the time when their mates are fertile. We measured urinary androgen levels and glucocorticoids in seven father tamarins for the first month following the birth of infants to determine 1) whether male tamarins showed an androgen response to their mate's postpartum ovulation, 2) when androgens rise relative to ovulation, 3) whether there is a glucocorticoid response, and 4) whether males alter their parenting behavior during their mate's receptive period. All of the males showed a significant increase in urinary androgens prior to the female's postpartum LH peak, which indicated ovulation. The hormonal increase, which included estradiol, occurred 3-7 days prior to the female's LH peak at a time that coincided with the female's follicular period. Corticosterone levels also peaked during that time, but did not correlate with androgen changes. Fathers did not alter their daily infant-carrying patterns relative to the androgen increase or at the time of the mate's LH peak. We conclude that male cotton-top tamarins experience an increase in androgens that coincides with their mate's postpartum ovulation, which ensures optimal fertility. However, this sexual communication does not alter father infant interactions, which already occur at a high rate in this species. PMID- 15356859 TI - Dominance rank and fecal testosterone levels in adult male chimpanzees (Pan troglodytes schweinfurthii) at Ngogo, Kibale National Park, Uganda. AB - In an attempt to describe hormone-behavior interactions in a sample of wild male chimpanzees, we quantified testosterone in 67 fecal samples obtained from 22 adult male chimpanzees at Ngogo, Kibale National Park, Uganda. A mixed-model methodology that controlled for age-class identified a significant positive association between testosterone levels and dominance rank. The results are consistent with those reported from a separate, smaller chimpanzee community in the same population in a study that analyzed testosterone levels in urine [Muller & Wrangham, 2004]. As in that earlier study, our results held during a period of social stability, which is not consistent with predictions of the "challenge hypothesis." We concur with Muller and Wrangham [2004] that the challenge hypothesis requires modification to explain the chimpanzee data, because fission fusion sociality in chimpanzees makes challenges unpredictable. We also discuss the utility of fecal samples and a mixed-model statistical method for behavioral endocrinology studies. PMID- 15356860 TI - Quantitative data on training new world primates to urinate. AB - This study assessed the effectiveness of operant conditioning in training three species of captive callitrichid primates (Leontopithecus rosalia, Callithrix geoffroyi, and Saguinus imperator) to urinate on demand. There were three goals to the study: 1) to develop a system for quantitatively assessing positive reinforcement training; 2) to ascertain whether or not positive reinforcement techniques can be used to train callitrichid monkeys to urinate on demand, and if so, how many training sessions are required; and 3) to determine the effect on urination behavior of the trainer entering the cage to collect a urine sample. Positive reinforcement with a continuous reinforcement schedule was used to capture a natural behavior: urination. Training sessions (30 min each) were conducted at dawn thrice weekly during five consecutive phases: habituation, control, training (animals were rewarded for urinating), maintenance (animals had reached a defined training criteria and continued to be rewarded for urinating), and collection (animals were rewarded for urinating, and the trainer entered the cage to collect the sample). The numbers of 30-min training sessions required to train the three monkey species (L. rosalia, C. geoffroyi, and S. imperator) were five, six, and eight, respectively. For the three species, the mean number of urinations per animal was significantly greater during the training, maintenance, and collection phases compared to the control phase. However, the three species differed significantly in the manner in which the rates of urination changed across the five phases. A higher proportion of subjects urinated during the training, maintenance, and collection phases compared to the control phase. Latency to first urination varied significantly across the five phases, with significantly reduced latencies to urinate during the training, maintenance, and collection phases compared to the control phase. The entry of the trainer into the cage to collect the urine sample did not appear to alter urination behavior. We demonstrate that operant conditioning techniques, which typically incur minimal cost, time investment, and disturbance, can be used to increase the quantity of urine samples collected for physiological analysis, the proportion of animals that urinate, and the speed of sample collection. PMID- 15356861 TI - Life-history correlates of steroid concentrations in wild peripartum baboons. AB - Steroid concentrations during late pregnancy and early lactation may be affected by both a female's reproductive history and her current condition, and may in turn predict subsequent life-history events, such as offspring survival. This study investigated these relationships in a wild primate population through the use of fecal steroid analysis in repeated sampling of peripartum baboons (Papio cynocephalus). Fecal samples were collected from 32 females in five groups within the Amboseli basin during 8 weeks prior to parturition and 13 weeks postpartum. From December 1999 through February 2002, 176 fecal samples were collected from individuals representing 39 peripartum periods. Fecal concentrations of progestins (fP), estrogen metabolites (fE), glucocorticoids (fGC), and testosterone metabolites (fT) were measured by radioimmunoassay. Steroid concentrations declined from late pregnancy to lactation, and the decline was greatest and most precipitous for fE and fP. Primiparous females had significantly higher mean fE concentrations in each of the last 2 months of pregnancy compared to multiparous females. Among multiparous females, fE and fT were significantly higher during late pregnancy in females carrying a male fetus compared to those carrying a female fetus. During early lactation, high fT in young mothers predicted subsequent infant death during the first year of life. These findings illustrate the potential power of repeated fecal-steroid sampling to elucidate mechanisms of life-history variability in natural populations. They also document significant differences in hormone profiles among subgroups, and highlight that such normative subgroup information is essential for interpreting individual variability in hormone-behavior associations. PMID- 15356862 TI - Reproductive output, maternal age, and survivorship in captive common marmoset females (Callithrix jacchus). AB - Common marmosets (Callithrix jacchus) demonstrate significant variation in reproductive output on both a yearly and lifetime basis in comparison to other anthropoid primates. We explore the factors that may be most important in determining reproductive variation in captive common marmosets. Studies have found that maternal age, maternal condition, and dam survivorship are related to reproductive output; however, these reports are not in agreement and are far from conclusive. With the use of a large, multicolony, demographic database pooling data across five marmoset colonies, we examined litter information for 1,649 litters, and reproductive summaries for 400 dams to assess 1) how reproductive output variation (total production, total weaned production) is determined by litter size, interbirth interval (IBI), age at first birth, and dam survival age; 2) the relationship between maternal age and reproductive output variables; and 3) relationship between the reproductive output variables and survival. We used stepwise regression procedures to describe the amount of variation in lifetime reproductive output among dams, and found that mean litter size accounted for 18% of the variance in total production, survival age accounted for 10.6%, age at first birth accounted for 8.8%, and mean IBI accounted for 5%. For total (nonzero) weaned production, survival age accounted for 7.6% of variance, age at first birth accounted for 7.2%, mean IBI accounted for 2%, and mean litter size accounted for 1.6%. We identified significant effects (P<0.05) of maternal age on litter size and IBI length, but no effect of dam age on weaned litter size. Cox proportional hazards regression analyses revealed significant effects (P<0.01) of number of litters, age at first birth, and site on dam survivorship. Dams that produced more litters showed higher survivorship. Age at first birth showed a positive relationship with dam survivorship, i.e., dams that delayed first reproduction had higher survival. Our findings about reproductive variation in marmosets may have practical applications for the management of marmoset breeding colonies. PMID- 15356863 TI - Reproductive parameters and life-history variables in captive golden-bellied mangabeys (Cercocebus agilis chrysogaster). AB - An understanding of the reproductive physiology of captive primates is vital for their successful management. We report on reproductive parameters and life history variables collected at the Sacramento Zoo for five female golden-bellied mangabeys (Cercocebus agilis chrysogaster) over a 7-year period. For each female, we collected data on their sexual skins, menses, gestation, and other pregnancy related factors, such as postconception bleeding and swelling. We used these data to estimate life-history variables, such as age at onset of estrus, menses, and conception, as well as the duration of intervals between births, and between parturition and the resumption of cycling. Cercocebus agilis chrysogaster is comparable to other Cercocebus species in terms of reproductive parameters, although variability is exhibited within and among female subjects. In some cases, it appears that stressful incidents altered cycle length or halted cycling altogether. We suggest the use of husbandry practices that include consistent data collection to enhance regularity in reproductive cycles, and maximize captive breeding success for this rare species. PMID- 15356864 TI - Putative implication of alpha-amylase loop 7 in the mechanism of substrate binding and reaction products release. AB - Alpha-amylases are widespread endo-enzymes involved in the hydrolysis of internal alpha-(1,4) glycosidic linkages of starch polymers. Molecular modeling of amylose amylase interactions is a step toward enzymatic mechanism understanding and rational design of new enzymes. From the crystallographic complex of barley alpha amylase AMY2-acarbose, the static aspects of amylose-amylase docking have been characterized with a model of maltododecaose (DP12) (G. Andre, A. Buleon, R. Haser, and V. Tran, Biopolymers 1999, Vol. 50, pp. 751-762; G. Andre and V. Tran, Special Publication no. 246 1999, The Royal Society of Chemistry, H. J. Gilbert, G. J. Davies, B. Henrissat, and B. Svensson, Eds., Cambridge, pp. 165-174). These studies, consistent with the experimental subsite mapping (K. Bak-Jensen, G. Andre, V. Tran, and B. Svensson, Journal of Biological Chemistry, to be published), propose a propagation scheme for an amylose chain in the active cleft of AMY2. The topographical overview of alpha-amylases identified loop 7 as a conserved segment flanking the active site. Since some crystallographic experiments suspected its high flexibility, its putative motion was explored through a robotic scheme, an alternate route to dynamics simulations that consume CPU time. The present article describes the characteristics of the flexibility of loop 7: location and motion in AMY2. A back-and-forth motion with a large amplitude of more than 0.6 nm was evaluated. This movement could be triggered by two hinge residues. It results in the loop flipping over the active site to enhance the docking of the native helical substrate through specific interactions, it positions the catalytic residues, it distorts the substrate towards its transition state geometry, and finally monitors the release of the products after hydrolysis. The residues involved in the process are now rational mutation points in the hands of molecular biologists. PMID- 15356865 TI - Intermolecular interaction studies of winter flounder antifreeze protein reveal the existence of thermally accessible binding state. AB - The physical nature underlying intermolecular interactions between two rod-like winter flounder antifreeze protein (AFP) molecules and their implication for the mechanism of antifreeze function are examined in this work using molecular dynamics simulations, augmented with free energy calculations employing a continuum solvation model. The energetics for different modes of interactions of two AFP molecules is examined in both vacuum and aqueous phases along with the water distribution in the region encapsulated by two antiparallel AFP backbones. The results show that in a vacuum two AFP molecules intrinsically attract each other in the antiparallel fashion, where their complementary charge side chains face each other directly. In the aqueous environment, this attraction is counteracted by both screening and entropic effects. Therefore, two nearly energetically degenerate states, an aggregated state and a dissociated state, result as a new aspect of intermolecular interaction in the paradigm for the mechanism of action of AFP. The relevance of these findings to the mechanism of function of freezing inhibition in the context of our work on Antarctic cod antifreeze glycoprotein (Nguyen et al., Biophysical Journal, 2002, Vol. 82, pp. 2892-2905) is discussed. PMID- 15356866 TI - Titration of poly(dA-dT) . poly(dA-dT) in solution at variable NaCl concentration. AB - CD and uv absorption data showed that high molecular weight poly(dA-dT) . poly(dA dT), at 298 K, undergoes an acid-induced transition from B-double helix to random coil in NaCl solutions of different concentrations, ranging from 0.005 to 0.600M. Similarly, titration of the polynucleotide with a strong base causes duplex-to single strands transition. The base- and acid-induced transitions were both reversible by back-titration (with an acid or, respectively, with a base): the apparent pKa were the same in both directions. However, the number of protons per titratable site (adenine N1) required to reach half-denaturation was in great excess over the stoichiometric value; to a much larger extent, the same effect was observed also for the deprotonation of the N3H sites of thymine. Moreover, in the basic denaturation experiments, at low salt concentrations ([NaCl]< or =0.300M) less acid than calculated was needed to back-titrate the base excess to half-denaturation. Both effects could be qualitatively justified on the basis of the counterion condensation theory of polyelectrolytes and considering the energy barrier created by the negatively charged phosphodiester groups to the penetration of the OH- ions inside the double helix and the screening effect of the Na+ ions on such charges, in the deprotonation experiments. PMID- 15356867 TI - Structural properties and photophysical behavior of conformationally constrained hexapeptides functionalized with a new fluorescent analog of tryptophan and a nitroxide radical quencher. AB - The influence of the conformational properties on the photophysics of two de novo designed hexapeptides was studied by spectroscopic measurements (ir, NMR, steady state, and time resolved fluorescence) and molecular mechanics calculations. The peptide sequences comprise two nonproteinogenic residues: a beta-(1-azulenyl)-L alanine (Aal) residue, obtained by formally functionalizing the Ala side chain with the azulene chromophore, and a Calpha-tetrasubstituted alpha-amino acid (TOAC), incorporating a nitroxide group in a cycloalkyl moiety. Aal represents a new fluorescent, quasi-isosteric Trp analog and TOAC a stable radical species, frequently used as a paramagnetic probe in biochemical studies. The peptide chains differ in the sequence position of the two probes and are heavily based on Aib (alpha-aminoisobutyric acid) residues to generate conformationally restricted helical structures, as confirmed by both spectroscopic and computational results. The conformationally controlled, excited state interactions, determining the photophysical relaxation of the Aal*/TOAC pair, are also discussed. PMID- 15356868 TI - CD studies on films of amyloid proteins and polypeptides: quantitative g-factor analysis indicates a common folding motif. AB - Irrespective of the constituent protein, all amyloid fibrils show similar morphology in the electron microscope and x-ray diffraction patterns characteristic of a "cross-beta" structure, with extended beta-strands perpendicular to the fibril's long axis. Little is known about the amount or type of this structure. I have measured CD spectra of films formed from a number of amyloid proteins and polypeptides, and estimated their contents of extended secondary structure, by analysis of their g-factor spectra, the ratio of the CD and absorbance signals (P. McPhie, Analytical Biochemistry, 2001, Vol. 293, pp. 109-119). Amyloid films of Abeta-(1-40) peptide, beta-2-microglobulin, insulin, and three homopolypeptides show very intense CD spectra, compatible with the presence of a beta-helix-like structure, arranged in a common framework in the fibrils. The extent of this structure was estimated as 45-80% in the protein fibrils and 30-80% in the polypeptide fibrils. PMID- 15356869 TI - The Mycobacterium tuberculosis chaperonin 10 monomer exhibits structural plasticity. AB - The conditions which favor dissociation of oligomeric Mycobacterium tuberculosis chaperonin 10 and the solution structure of the monomer were studied by analytical ultracentrifugation, size exclusion chromatography, fluorescence, and circular dichroism spectroscopies. At neutral pH and in the absence of divalent cations, the protein is fully monomeric below approximately a 4.7 microM concentration. Under these conditions the monomer forms completely unfolded and partially folded conformers which are in equilibrium with each other. One conformer accumulates over the others which is stable within a very narrow range of temperatures. It contains a beta-sheet-structured C-terminal half and a mostly disordered N-terminal half. Other components of the equilibrium include partially helical structures which do not completely unfold at high temperature or under strong acidic conditions. Complete unfolding of the monomer occurs in the presence of denaturants or below 14 degrees C. Cold-denaturation is detected at an unusually high temperature and this may be due to the concentration of hydrophobic residues, which is larger in chaperonins than in other globular proteins. Finally, the monomer self-associates in the pH range 5.8-2.9, where it forms small oligomers. A structure-activity relationship was investigated with the sequences known to be involved in the various biological activities of the monomer. PMID- 15356870 TI - Determining beta-sheet stability by Fourier transform infrared difference spectra. AB - We describe here a new method for determining the conformational stability of antiparallel beta-sheets. Due to coupling between the transition dipoles, beta sheet conformations typically exhibit a characteristic high-frequency amide I component centered at approximately 1680 cm(-1). Using one beta-sheet protein and two small beta-hairpins, we demonstrate that this high-frequency component, which is fairly narrow (approximately 8-10 cm(-1)), can be quantitatively resolved and used in thermal stability determination. Compared with the commonly used CD and fluorescence techniques, this ir method offers advantages. Since the area of this high-frequency component is only proportional to the folded population, it eliminates the need for a priori information of the folded and unfolded baselines encountered in other methods. Thus, it is applicable to a variety of beta-sheet systems. PMID- 15356871 TI - Conformational states of HRPA1 induced by thermal unfolding: effect of low molecular weight solutes. AB - Fluorescence, CD, and activity measurements were used to characterize the different conformational states of horseradish peroxidase A1 induced by thermal unfolding. Picosecond time-resolved fluorescence studies showed a three exponential decay dominated by a picosecond lifetime component resulting from energy transfer from tryptophan to heme. Upon thermal unfolding a decrease in the preexponential factor of the picosecond lifetime and an increase in the quantum yield were observed approaching the characteristics observed for apoHRPA1. The fraction of heme-quenched fluorophore decreased to 0.4 after unfolding as shown by acrylamide quenching. A new unfolding pathway for HRPA1 was proposed and the effect of the low molecular weight solutes trehalose, sorbitol, and melezitose on this pathway was analyzed. Native HRPA1 unfolds with an intermediate between the native and the unfolded conformation. The unfolded conformation can refold to the native state or to a native-like conformation with no calcium ions upon cooling or can give an irreversible denatured state. The refolded conformation with no calcium ions was clearly identified in a second thermal scan in the presence of EDTA and shows secondary and tertiary structures, heme reincorporation in the cavity, and at least 59% of activity. Melezitose stabilized the refolded Ca2+ depleted protein and induced a more complex mechanism for heme disruption. The effect of sorbitol and trehalose were mainly characterized by an increase in the temperature of unfolding. PMID- 15356872 TI - Morphologies and conformation transition of lentinan in aqueous NaOH solution. AB - Molecular morphologies and conformation transition of lentinan, a beta-(1-->3)-D glucan from Lentinus edodes, were studied in aqueous NaOH solution by atomic force microscopy (AFM), viscometry, multiangle laser light scattering, and optical rotation measurements. The results revealed that lentinan exists as triple-helical chains and as single random-coil chains at NaOH concentration lower than 0.05M and higher than 0.08M, respectively. Moreover, the dramatic changes in weight-average molecular weight Mw, radius of gyration [s2](1/2), intrinsic viscosity [eta], as well as specific optical rotation at 589 nm [alpha]589 occurred in a narrow range of NaOH concentration between 0.05 and 0.08M NaOH, indicating that the helix-coil conformation transition of lentinan was carried out more easily than that of native schizophyllan and scleroglucan, and was irreversible. For the first time, we confirmed that the denatured lentinan molecule, which was dissolved in 0.15M NaOH to be disrupted into single coil chains, could be renatured as triple helical chain by dialyzing against abundant water in the regenerated cellulose tube at ambient temperature (15 degrees C). In view of the AFM image, lentinan in aqueous solution exhibited the linear, circular, and branched species of triple helix compared with native linear schizophyllan or scleroglucan. PMID- 15356873 TI - Dynamics of water molecules buried in cavities of apolipoprotein E studied by molecular dynamics simulations and continuum electrostatic calculations. AB - Molecular dynamics (MD) simulations of several nanoseconds each were used to monitor the dynamic behavior of the five crystal water molecules buried in the interior of the N-terminal domain of apolipoprotein E. These crystal water molecules are fairly well conserved in several apolipoprotein E structures, suggesting that they are not an artifact of the crystal and that they may have a structural and/or functional role for the protein. All five buried crystal water molecules leave the protein interior in the course of the longest simulations and exchange with water molecules from the bulk. The free energies of binding evaluated from the electrostatic binding free energy computed using a continuum model and estimates of the binding entropy changes represent shallow minima. The corresponding calculated residence times of the buried water molecules range from tens of picoseconds to hundreds of nanoseconds, which denote rather short times as for buried water molecules. Several water exchanges monitored in the simulations show that water molecules along the exit/entrance pathway use a relay of H bonds primarily formed with charged residues which helps either the exit or the entrance from or into the buried site. The exit/entrance of water molecules from/into the sites is permitted essentially by local motions of, at most, two side chains, indicating that, in these cases, complex correlated atomic motions are not needed to open the buried site toward the surface of the protein. This provides a possible explanation for the short residence times. PMID- 15356874 TI - Comparison of conventional tube test technique and gel microcolumn assay for direct antiglobulin test: a large study. AB - Gel microcolumn assay (GMA) is a modified serological technique that has been used for ABO and Rh typing, direct antiglobulin test (DAT), detecting alloantibodies, red cell phenotyping, and other applications. However, for DAT, the role of GMA is controversial. The purpose of this large study was to compare the performance of the conventional tube test (CTT) to GMA for detecting potentially significant antibodies coating red blood cells in vivo. From January 1996 to May 2002, we performed DATs by GMA and CTT on 9,862 blood samples submitted to our reference laboratory, using LISS/Coombs cards (DiaMed-Latino America, Lagoa Santa-MG, Brazil) for GMA and polyspecific and monospecific anti IgG reagents for CTT. Acid eluates were prepared from all positive DAT samples. The specificity of eluates was determined by GMA. We detected nonconcordant results in 2,079 out of 3,163 positive DATs (65.7%). All of these tests were only positive in GMA. Sensitivity and specificity for DATs was 100% and 83.0% for gel, and 50.7% and 97.8% for tube, respectively. Based on this study GMA showed to be more sensitive than CTT for detecting potentially significant antibodies coating red blood cells in vivo. PMID- 15356875 TI - Pronto ThromboRisk--a novel primer-extension ELISA based assay for the detection of mutations associated with increased risk for thrombophilia. AB - The role of inherited thrombophilias in the pathogenesis of thrombosis, and their relation to thromboembolic events and pregnancy complications, has escalated the demand for molecular testing. We employed the new Pronto ThromboRisk kit (Pronto Diagnostics Ltd., Rehovot, Israel), which is based on a novel primer-extension ELISA assay, for the simultaneous detection of three genetic mutations. These are Factor V Leiden G1691A (R506Q), prothrombin G20210A, and methylenetetrahydrofolate reductase (MTHFR) C677T, which are known to have increased incidence in patients with thromboembolic events and pregnancy complications. We examined 284 randomly selected patient samples from the special coagulation hospital laboratory. The results using the new assay were compared to those obtained by routinely employed conventional molecular techniques. The ThromboRisk kit provided identical results with no false-negative results and with a high specificity of over 99% for all three mutations. We conclude that the Pronto ThromboRisk kit allows fast, precise, and reliable testing for the three genetic mutations. The assay is easy to perform and provides a useful tool for screening high risk populations, such as patients with a personal or family history of venous thromboembolism, women with pregnancy complications, or users of oral contraceptives. PMID- 15356876 TI - Rapid detection of Bordetella pertussis by real-time PCR using SYBR green I and a LightCycler instrument. AB - A polymerase chain reaction (PCR) assay in real-time for detection of B. pertussis using SYBR green I as the reporter fluorophore and LightCycler instrument (a thermocycler coupled to a fluorescence detection device) was established and evaluated. The amplified amplicon using series diluted control prototype strain (ATCC strain #9797) of B. pertussis was analyzed for the fluorescent melting profile, and melting temperature (Tm) was determined. When examined, amplicons using a representative set of clinical isolates of B. pertussis were found to have the same Tm value (86 +/- 0.5 degrees C, the specificity parameter of detection) as the control prototype strain as expected. Amplified product was also analyzed and detected by agarose gel electrophoresis. The detection limit by fluorescent profile and Tm analysis was 10-fold better than that detected by agarose gel analysis. PMID- 15356877 TI - Simultaneously multi-parameter determination of hematonosis cell apoptosis by two photon and confocal laser scanning microscopy. AB - Flow cytometry (FCM), TdT-mediated dUTP Nick End Labeling (TUNEL), and DNA ladder are conventional methods to detect apoptosis of drug-treated cells. However, the assumption of cell number restricts their applications in clinics. In this paper, we report a cell-saving imaging method for quick identification of the hematonosis cell apoptosis induced by chemotherapeutic drugs. By the combination of two-photon and confocal microscopy, three main apoptosis parameters (the change of nuclear morphology, collapse of mitochondrial membrane potential, and increase of intracellular calcium) were recorded simultaneously for single As2O3 induced Molt-4 cells. The results are highly in accordance with those produced by classical flow cytometry. This work suggests that this new imaging method would be promising in the quick identification of hematonosis cell apoptosis. PMID- 15356878 TI - Urokinase gene 3'-UTR T/C polymorphism is associated with oral cancer. AB - Urokinase is thought to be involved in the formation of oral cancer, although there is a lack of genetic evidence. Our aim was to study single nucleotide polymorphisms in order to investigate the possibility. A total of 130 oral cancer patients and 105 controls were studied. Polymerase chain reaction (PCR) based restriction analysis was used to identify the C/T polymorphism of the urokinase gene, which is located on the 3'-untranslated region (3'-UTR) of chromosome 10. There was a significant difference in the distribution of the urokinase gene 3' UTR C/T polymorphism frequency between cancer patients and the normal control group (P < 0.05). The "T" allele was prominent in the cancer group. The odds ratio for the risk of the "T" allele in cancer patients was 2.71 (95% CI = 1.325 approximately 5.562). The cancer patients were further categorized according to gender and whether or not they were habitual smokers or betel nut chewers. These clinical parameters were then compared with tumor cell differentiation and tumor progression. No significant differences were found. Therefore, the urokinase gene 3'-UTR "T" allele is associated with oral cancer and may play a role in oral cancer formation. However, we did not find the relationship between tumor progression and this polymorphism. PMID- 15356879 TI - Measurement of C-reactive protein: two high sensitivity methods compared. AB - C-reactive protein (CRP) is an acute phase marker and a predictor of the risk of developing atherosclerotic complications. However, as a predictor of this risk, high sensitivity measurements are needed, and high sensitive CRP (hsCRP) assays have been developed. In this study, we experimentally compared two hsCRP assays, based on nephelometry and turbidimetry, both implemented on automated analyzers. Linearity, imprecision, turbidity interference, and results in the assay of 96 samples have been compared. Method comparison of the same two analytical systems in the assay of CRP was also performed on the basis of results in an interlaboratory external quality assessment scheme (EQAS). The two systems were found to perform substantially equally, both in hsCRP and in CRP measurement, but in the hsCRP assay the precision of nephelometry (CV% in the interval 3.0-5.8) was lower than that of turbidimetry (CV% in the interval 1.8-2.3). The classification of results by the two methods into three predefined relative risk classes gave 18% rate of discordance, in any case by one class only. The two methods proved reliable and comparable in the measurement of hsCRP, but precision should be improved. PMID- 15356880 TI - 150th Anniversary of John Snow and the pump handle. PMID- 15356882 TI - Not cloning: new embryo research has sparked controversy. PMID- 15356883 TI - Don't ignore stroke warning sign. PMID- 15356884 TI - Clues to chronic fatigue syndrome. PMID- 15356886 TI - Ending end-of-life care: who decides? PMID- 15356887 TI - Bone marrow transplants and breast cancer. PMID- 15356885 TI - What are allergies? PMID- 15356888 TI - Nutritional deficiencies among the elderly. PMID- 15356889 TI - Jury out on estrogen and Alzheimer's. PMID- 15356890 TI - "Complementary" therapies may compete with prostate cancer treatment. PMID- 15356891 TI - Marked for heart trouble? PMID- 15356892 TI - Beta-blockers and heart failure. PMID- 15356893 TI - New drug for bothersome bowels. PMID- 15356894 TI - Meditation helps arteries. PMID- 15356895 TI - Questions and answers. I've heard that magnets can control pain. Is this true? PMID- 15356896 TI - Questions and answers. When does a person with a cold spread germs--before symptoms appear or after sneezing and sniffling begin? PMID- 15356897 TI - Questions and answers. What are safe and practical exercises for someone with arthritis? PMID- 15356898 TI - [The crankshaft phenomenon]. PMID- 15356899 TI - [Thoracoscopic surgery: current status and prospects]. PMID- 15356900 TI - [Progress on the study of genes associated with individual susceptibility to noise-induced hearing loss]. PMID- 15356901 TI - Easing the pain. PMID- 15356903 TI - Radiographic criteria for placement of translaminar facet screws. AB - BACKGROUND CONTEXT: Screw fixation of the facet joint has been reported to stabilize the lumbar spine and facilitate spinal fusion. Accurate placement of translaminar facet screws (TLFSs) requires identification of the posterior spinal elements, and the facet joints in particular, which may be facilitated by intra operative fluoroscopy. PURPOSE: The purpose of this study was to define the radiographic parameters that will allow for successful placement of TLFSs. STUDY DESIGN: Eighteen TLFSs were placed in three human cadaver spines using fluoroscopic guidance, with screws placed from L3-L4 to L5-S1 bilaterally. After screw placement, the spines were dissected and examined for accuracy of screw placement. METHODS: In the first cadaver spine, TLFSs were placed with direct visualization of the posterior lumbar spine, during which a fluoroscopic image intensifier was used to define the appropriate radiographic parameters for safe placement of the screws. Bilateral TLFSs were then placed percutaneously using the radiographic parameters developed, after which the spines were dissected to examine the positions of the screws. RESULTS: The radiographic views identified to achieve proper TLFS placement were a true lateral, anteroposterior (AP), a 45 degree oblique and an AP view with the X-ray bean at 30- to 45-degree cephalad angle ("spinal outlet" view). Using these views and the defined radiographic criteria, proper positioning of percutaneously placed TLFSs was achieved, with no spinal canal breaches found. CONCLUSIONS: This study defines intra-operative radiographic criteria that will assist in placement of TLFSs. The use of this technique may allow for screw placement with less extensive exposure of the posterior spine. PMID- 15356902 TI - Cellular proliferation in the cerebral cortex following neural excitation in rats. AB - Cortical spreading depression (SD) is characterized by propagation of neuronal/glial membrane depolarization throughout the unilateral cerebral cortex and has been linked to several neurological disorders, including migraine aura and epilepsy. SD induction resulted in a dramatic increase in BrdU-incorporated cells in the ipsilateral cortical hemisphere that was dependent on the number of elicited SD. Immunohistochemical studies revealed that 53% of the BrdU-labeled cells in the SD-generated cortex were NG2 immunopositive and 25% were OX-42 immunopositive. The remaining 22% of BrdU-incorporated cells showed no immunoreactivity to GST-rr, GFAP, NeuN, NG2 or OX-42. These data indicate that functional excitation of the cerebral cortex induces proliferative response in cortical cells, which may subsequently differentiate into glial progenitor or microglia within 3 days after stimulation. PMID- 15356904 TI - Some usability testing resources. PMID- 15356905 TI - Speaking out for children. Interview by Janis Smy. PMID- 15356906 TI - Genomic DNA detection using cycling probe technology and capillary gel electrophoresis with laser-induced fluorescence. AB - Cycling probe technology (CPT) is an isothermal DNA analysis method that has been shown to be useful for identifying genetic markers of antibiotic-resistant bacteria in clinical settings. CPT assays have previously employed several assay methods that include polyacrylamide gel electrophoresis and magnetic beads for separations and radioisotopic and colorimetric detection for detection. Capillary gel electrophoresis with laser-induced fluorescence (CGE-LIF) is an alternative separation and detection method that offers attributes such as low sample consumption, short analysis times, no radiation hazards and potential for high throughput. We report on the development of a CGE-LIF CPT assay for genomic DNA from Erwinia herbicola and the comparison of this assay with a conventional 32p radioisotopic PAGE CPT assay. Separation and detection of intact and cleaved fluorescein-labeled CPT probe molecules by CGE-LIF was achieved in under 4 min through a gel-filled capillary (7 cm separation length to detector). Total time, from setup to detection, was about 1 h for the complete assay versus several hours (3-12 h) for the radioisotopic PAGE CPT assay. Similar detection limits of 10(5)-10(6) copies of genomic target DNA were observed with each assay method. This study demonstrated that CGE-LIF CPT is a suitable analysis method for the detection of genomic DNA sequences. PMID- 15356907 TI - A pioneering primary care nursing service. PMID- 15356908 TI - Value in global standardization of outbred rats for regulatory pharmaceuticals studies. AB - As a result of a revolution in globalization over the last 10 years, few pharmaceutical companies conduct their non-clinical studies within a single region and virtually none conduct all non-clinical studies in house. The increased activity of product in-licensing at all stages of development results in further segmentation of toxicological source information. Juxtaposed to this segmental collection of toxicology information is the nature of toxicological testing for pharmaceuticals, an iterative process, carefully building subsequent testing designs and analyses upon the foundation of previously identified outcomes and associated safety issues. As a consequence of the segmentation of global product development, today's analyses of toxicological outcomes for single pharmaceutical projects often involve a conglomeration of results from studies conducted in several world regions using a variety of sources of animals. Such practices can lead to discordant study results and difficulty in understanding or rationalizing a compound's toxicological and pharmacological profile, and eventually how this relates to human risk. Despite the variety in source information, an integration of the total toxicology/pharmacology data must be made by industry, and/or by health authorities. To improve this integration, the development and use of better standardized genetics for rodent species used in pharmaceutical testing is more imperative today than in the past, as a result of a constellation of changes in industry policies. PMID- 15356909 TI - Peripheral arterial disease--towards extended awareness and improved prevention of ischaemic events. PMID- 15356910 TI - Providing holistic sickle cell care. PMID- 15356911 TI - Making a mark in dermatology. Interview by Alison Whyte. PMID- 15356912 TI - Where intraoperative reflexes can be used to monitor nerve root and spinal cord gray matter function. PMID- 15356913 TI - Nifedipine GITS: perspectives beyond blood pressure. PMID- 15356914 TI - PERSUADE...(a EUROPA sub-study)...results: perindopril protects diabetic patients with coronary disease from cardiovascular death and heart attacks. PMID- 15356915 TI - EUROPA: clinical benefits, clinical promise. PMID- 15356916 TI - CCR5 targeted SIV vaccination strategy preventing or inhibiting SIV infection. AB - Cell-surface CCR5 is a major coreceptor with CD4 glycoprotein, mediating cellular entry of CCR5 strains of HIV-1 or SIV. We targeted the SIV CCR5 coreceptor in a combined CCR5-SIV antigen immunization strategy. Rhesus macaques were immunized i.m. with the 70 kDa heat shock protein (HSP70) covalently linked to the CCR5 peptides, SIV gpl20 and p27. Intravenous challenge with SIV mac 8980 prevented SIV infection or decreased the viral load with the CCR5-SIV combined vaccine. CC chemokines and antibodies which block and downmodulateCCR5 were induced, as well as immune responses to the subunit SIV antigens. This novel vaccination strategy complements cognate immunity to SIV with innate immunity to the CCR5 coreceptor of SIV. PMID- 15356917 TI - Protective effect of macrocyclic binuclear oxovanadium complex on oxidative stress in pancreas of streptozotocin induced diabetic rats. AB - Chronic hyperglycemia in diabetes is a major causative factor of free radical generation which further leads to many secondary diabetic complications via the damage to cellular proteins, membrane lipids, nucleic acids and eventually to cell death. Recently we have reported on the hypoglycemic efficacy of a new macrocyclic binuclear oxovanadium complex and its non-toxic nature. This study focuses on the effect of the above complex in ameliorating oxidative stress in the pancreas of diabetic rats. Streptozotocin induced diabetic rats were treated orally with the vanadium complex (5 mg/kg/body weight) for 30 days and the level of pancreatic antioxidants and lipid peroxides were determined. Treatment with the macrocyclic binuclear oxovanadium complex decreased the lipid peroxides and the antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase to near control levels. Histological examinations also revealed the protective effect of the complex on pancreatic beta cells. The results demonstrate the protective effect of the macrocyclic binuclear oxovanadium complex on the pancreatic antioxidant status. PMID- 15356918 TI - Stimulation of endothelial IL-8 (eIL-8) production and apoptosis by phenolic metabolites of benzene in HL-60 cells and human bone marrow endothelial cells. AB - Benzene toxicity is considered to be elicited by its metabolites and phenolic metabolites of benzene are known to induce apoptosis in leukemia cells in culture and in human bone marrow progenitor cells. One potential mechanism of apoptosis induced by benzene metabolites that has not been examined is the production of pro-apoptotic cytokines such as endothelial IL-8 from endothelial cells in bone marrow stroma. In this study, we utilized HL-60 cells which are known to produce the endothelial form of IL-8 (eIL-8) and human bone marrow endothelial cells (HBMEC) as model systems. Hydroquinone (HQ), Catechol (Cat) and benzenetriol (BT) all induced eIL-8 production and apoptosis in HL-60 cells. HQ induced a marked 50 70 fold stimulation of eIL-8 levels and HL-60 cells were shown to have the eIL-8 receptor, CXCR1 thus enabling an autocrine pathway of apoptosis. However, treatment with recombinant eIL-8 failed to induce apoptosis in HL-60 cells as previously reported and antibodies to either IL-8 or CXCR1 did not significantly abrogate benzene metabolite-induced apoptosis. HQ and Cat but not BT also induced stimulation of eIL-8 production in HBMEC. These data demonstrate that although metabolites of benzene induce marked stimulation of eIL-8, this is unlikely to be responsible for apoptosis induced in HL-60 cells. Our data also demonstrates that phenolic metabolites of benzene stimulate the production of eIL-8 from HBMEC suggesting that higher levels of endothelial-derived cytokines may occur in bone marrow after benzene exposure. PMID- 15356919 TI - Mechanism of metal-mediated DNA damage induced by a metabolite of carcinogenic acetamide. AB - Acetamide is carcinogenic in rats and mice. To clarify the mechanism of carcinogenesis by acetamide, we investigated DNA damage by and acetamide metabolite, acetohydroxamic acid (AHA), using 32P-5'-end-labeled DNA fragments. AHA treated with amidase induced DNA damage in the presence of Cu(II) and displayed a similar DNA cleavage pattern of hydroxylamine. DNA damage was inhibited by both catalase and bathocuproine, suggesting that H2O2 and Cu(I) are involved. Carboxy-PTIO, a specific scavenger of nitric oxide (NO), partially inhibited DNA damage. The amount of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) by amidase-treated AHA was similar to that by hydroxylamine. ESR spectrometry revealed that amidase-treated AHA as well as hydroxylamine generated NO in the presence of Cu(II). From these results, it has been suggested that AHA might be converted into hydroxylamine by amidase. These results suggest that metal mediated DNA damage mediated by amidase-catalyzed hydroxylamine generation plays an important role in the carcinogenicity of acetamide. PMID- 15356920 TI - Reducing bioassay variability by identifying sources of variation and controlling key parameters in assay protocol. AB - Reducing bioassay variability by identifying sources of variation and controlling important parameters in assay protocol was demonstrated in this study. The variability of a bioassay based on a luminescent bacterium was examined as an example. This assay involved the growth of cells, storage at a low temperature, activation, and exposure to a test sample, and the assay response was bioluminescence inhibition. After determining that measurement error was small and negligible, the total assay variability was decomposed in an initial variance components study into between-batch, between-vial, and between-tube variations. Results indicated that between-vial variations accounted for the majority of the total observed variability and that reducing this type of variation would be beneficial. Five parameters in the assay protocol were determined as factors that potentially affected assay variability significantly. A split-plot design was employed to investigate the effects of these factors and some of their interactions on the assay response. One of the five factors, i.e., activation temperature, turned out to have a significant effect. The variance components study was repeated with better control of activation temperature as well as other parameters. Results indicated that the total variability of the bioassay was reduced by approximately 85%. PMID- 15356921 TI - Squamous odontogenic tumor versus pseudoepitheliomatous hyperplasia. PMID- 15356922 TI - Metabolism of chlorambucil by rat liver microsomal glutathione S-transferase. AB - Clinical efficacy of alkylating anticancer drugs, such as chlorambucil (4-[p-[bis [2-chloroethyl] amino] phenyl]-butanoic acid; CHB), is often limited by the emergence of drug resistant tumor cells. Increased glutathione (gamma glutamylcysteinylglycine; GSH) conjugation (inactivation) of alkylating anticancer drugs due to overexpression of cytosolic glutathione S-transferase (GST) is believed to be an important mechanism in tumor cell resistance to alkylating agents. However, the potential involvement of microsomal GST in the establishment of acquired drug resistance (ADR) to CHB remains uncertain. In our experiments, a combination of lipid chromatography/electrospray ionization mass spectrometry (LC/ESI/MS) was employed for structural characterization of the resulting conjugates between CHB and GSH. The spontaneous reaction of 1mM CHB with 5 mM GSH at 37 degrees C in aqueous phosphate buffer for 1 h gave primarily the monoglutathionyl derivative, 4-[p-[N-2-chloroethyl, N-2-S-glutathionylethyl] amino]phenyl]-butanoic acid (CHBSG) and the diglutathionyl derivative, 4-[p-[2-S glutathionylethyl] amino]phenyl]-butanoic acid (CHBSG2) with small amounts of the hydroxy-derivative, 4-[p-[N-2-S-glutathionylethyl, N-2-hydroxyethyl] amino]phenyl]-butanoic acid (CHBSGOH), 4-[p-[bis[2-hydroxyethyl] amino]phenyl] butanoic acid (CHBOH2), 4-[p-[N-2-chloroethyl, N-2-S-hydroxyethyl]amino]phenyl] butanoic acid (CHBOH). We demonstrated that rat liver microsomal GST presented a strong catalytic effect on these reactions as determined by the increase of CHBSG2, CHBSGOH and CHBSG and the decrease of CHB. We showed that microsomal GST was activated by CHB in a concentration and time dependent manner. Microsomal GST which was stimulated approximately two-fold with CHB had a stronger catalytic effect. Thus, microsomal GST may play a potential role in the metabolism of CHB in biological membranes, and in the development of ADR. PMID- 15356923 TI - A poisoned field: farmworkers, pesticide exposure, and tort recovery in an era of regulatory failure. PMID- 15356924 TI - Toward safer fields: using AWPA's working arrangement provisions to enforce health and safety regulations designed to protect farmworkers. PMID- 15356925 TI - Accumulation and tissue distribution of selected polychlorinated biphenyl congeners in chickens. AB - During one to three consecutive periods of 2 weeks, broiler chickens (n = 108) received test dies to which different amount of PCBs (7 congeners) were added. The relationship between exposure time and accumulation of individual congeners in different chicken tissues, such as breast, thigh and abdominal fat tissue, was observed. In all tissues, the vast majority of the PCB accumulation occurred during the first 2 weeks of exposures. After that, PCB concentrations only increased in the abdominal fat tissue of the animals. The individual PCBs were distributed differently in the various tissues. While CBs 28, 118, 138, 153 and 180 accumulated in the chickens, CBs 52 and 101 were metabolized, but no methyl sulphone metabolites of these congeners could be detected. Our results provide information on the absorption, tissue distribution and biotransformation of the individual PCB congeners and confirm the structure-activity relationships for metabolism of PCBs in birds, which are different from those in fish or mammalian species. PMID- 15356926 TI - Death by mosquito. Malaria, like AIDS, is killing millions. But unlike AIDS, it can be cured. Why isn't that happening? PMID- 15356927 TI - The kids are all right. Some are overweight and too many are poor, but by most other measures, U.S. children are doing great. PMID- 15356928 TI - How low can you go? New guidelines call for heart patients to cut their cholesterol levels even further. PMID- 15356929 TI - Changing history--closing the gap in AIDS treatment and prevention. PMID- 15356930 TI - Prevention of congenital syphilis--time for action. PMID- 15356931 TI - Economic and programmatic aspects of congenital syphilis prevention. AB - A review of the economic and programmatic aspects of congenital syphilis was conducted and recommendations made for improvement of its prevention. Congenital syphilis is a preventable disease and the tools to prevent it have been available for decades. In both industrialized and developing countries, but particularly the latter, the prevention of congenital syphilis by antenatal screening is cost effective and may be cost-saving. Yet, globally, there are probably >500 000 fetal deaths a year from congenital syphilis, a figure rivalling that from mother to-child transmission of human immunodeficiency virus (HIV), which receives far greater attention. The reasons that congenital syphilis persists vary, with international and national under-appreciation of the burden of congenital syphilis and insufficient political will to provide effective antenatal screening programmes probably being the main reasons. All causes are amenable to effective intervention programmes. The prevention of congenital syphilis should be a global priority; international agencies and national programmes should be committed to improving antenatal care (ANC) services including syphilis detection and prevention. PMID- 15356932 TI - Maternal and congenital syphilis programmes: case studies in Bolivia, Kenya and South Africa. AB - Preventing congenital syphilis is not technically difficult, however operational difficulties limit the effectiveness of programmes in many settings. This paper reports on programmes in Bolivia, Kenya, and South Africa. All three countries have established antenatal syphilis control programmes. Early antenatal syphilis screening and management of positive cases were difficult to implement since most women presented for their first antenatal clinic visit after 6 months of pregnancy. Most women had rapid plasma reagin (RPR) testing; results were available on the same day in some clinics but took up to 4 weeks in others. No clinic had a system for tracking RPR-reactive women who did not return for their results. There were no guidelines for providers in Kenya and Bolivia. In all countries, supplies, drugs, notification cards, and other consumables were often unavailable. Health-care providers were unmotivated in Kenya and reported an excessive client load. In South Africa and Kenya some clients reported at their exit interview that they had never heard of syphilis nor had they been informed why blood was collected. Several prevention strategies could be implemented at the clinic level. These include encouraging women to attend for antenatal care before the fourth month of pregnancy, providing point-of-care testing so that results are available immediately and women who test positive can be treated, implementing presumptive treatment of sexual partners of women who test positive, adding a second test later in pregnancy so that incident cases can be managed, and improving the quality of syphilis care during pregnancy, delivery, and the neonatal period. PMID- 15356933 TI - Antenatal syphilis control: people, programmes, policies and politics. AB - Antenatal syphilis control is an integral component of reproductive health policies in most countries. In many of these countries, however, the existence of a health policy does not automatically translate into an effective health programme. We argue that neglecting to take into account the perspectives of all stakeholders when planning programmes may be the reason that functional and sustained interventions for antenatal syphilis are lacking. Stakeholders may include health policy decision-makers, programme managers, service delivery personnel (on whom implementation depends), as well as the pregnant women, families, and communities who will most benefit from the intervention. We describe how to undertake a multilevel assessment in order to identify stakeholders, identify interlinked perspectives, and analyse these perspectives within the socioeconomic, cultural and political environment within which an intervention is designed to be delivered. Using this multidisciplinary approach, we propose that the barriers to, and opportunities for, turning health policy into effective practice will be identified, and the result will be the formulation of a broad programme response to ensure implementation of the policy. Undertaking a multilevel assessment is but the first step in identifying barriers to successful programmes. Currently there is a lack of strong political support for this intervention at national and international levels. Devising strategies to address these potential barriers requires a broad range of skills and approaches some of which are outlined in this paper. PMID- 15356934 TI - The prevention and management of congenital syphilis: an overview and recommendations. AB - The continued occurrence of congenital syphilis is an indictment of the inadequate antenatal care services and poor quality of programmes to control sexually transmitted infections. More than 1 million infants are born with congenital syphilis each year. Despite national policies on antenatal testing and the widespread use of antenatal services, syphilis screening is still implemented only sporadically in many countries, leaving the disease undetected and untreated among many pregnant women. The weak organization of services and the costs of screening are the principal obstacles facing programmes. Decentralization of antenatal syphilis screening programmes, on-site testing and immediate treatment can reduce the number of cases of congenital syphilis. Antenatal syphilis screening and treatment programmes are as cost effective as many existing public health programmes, e.g. measles immunization. Diagnosis of congenital syphilis is problematic since more than half of all infants are asymptomatic, and signs in symptomatic infants may be subtle and nonspecific. Newer diagnostic tests such as enzyme immunoassays, polymerase chain reaction and immunoblotting have made diagnosis more sensitive and specific but are largely unavailable in the settings where they are most needed. Guidelines developed for better-resourced settings are conservative and err on the side of overtreatment. They are difficult to implement in, or inappropriate for, poorly-resourced settings because of the lack of investigative ability and the pressure on health facilities to discharge infants early. This paper offers recommendations for treating infants, including an approach based solely on maternal serological status and clinical signs of syphilis in the infant. PMID- 15356935 TI - Controlling congenital syphilis in the era of HIV/AIDS. PMID- 15356936 TI - Maternal syphilis: pathophysiology and treatment. AB - Despite the long history of medical interest in syphilis and its effects on pregnancy outcome, many fundamental questions about the pathophysiology and treatment of syphilis during pregnancy remain unanswered. However, understanding has been advanced by recent scientific reports such as those which delineate the complete sequence of the genome of the syphilis spirochaete, provide a more precise description of fetal and neonate infection by use of rabbit infectivity tests and describe the gestational age distribution of fetal death secondary to syphilis. It appears that fetal syphilitic involvement progresses in a rather predictable fashion, and although there is disagreement about the optimal prenatal treatment regimen, programmatic efforts to prevent fetal death must provide seropositive pregnant women with a recommended treatment early in pregnancy, and certainly before the third trimester. PMID- 15356937 TI - Diagnostic tools for preventing and managing maternal and congenital syphilis: an overview. AB - Syphilis is a major cause of adverse outcomes in pregnancy in developing countries. Fetal death and morbidity due to congenital syphilis are preventable if infected mothers are identified and treated appropriately by the middle of the second trimester. Most pregnant women with syphilis are asymptomatic and can only be identified through serological screening. Non-treponemal tests, such as the rapid plasma reagin (RPR) test, are sensitive, simple to perform, and inexpensive. However, they have often not been available at primary health-care settings because they required cold storage for reagents and electricity to operate a rotator. Additionally, as many as 28% of positive RPR results in pregnant women are biological false positives. Confirmatory assays are usually available only in reference laboratories. Technological advances have resulted in improved serodiagnostic tools for syphilis. New enzyme immunoassays are available for surveillance and for large-scale screening programmes. Decentralized antenatal screening with on-site confirmation is now possible since new RPR reagents that are stable at room temperature have become commercially available, as have solar-powered rotators and simple, rapid point-of-care treponemal tests that use whole blood and do not require electricity or equipment. These will be valuable tools for preventing or eliminating congenital syphilis. The development of a non-invasive rapid treponemal test that distinguishes between active and past infections remains a high priority in areas where syphilis is endemic. PMID- 15356938 TI - Genital herpes and human immunodeficiency virus: double trouble. AB - The synergistic relationship between herpes simplex virus type 2 (HSV-2) and transmission of human immunodeficiency virus (HIV) can be substantial in developing countries that have high prevalences of both viral infections. Genital herpes, most frequently caused by HSV-2, has become the leading cause of genital ulcer disease worldwide. This review of recent research on genital herpes and enhanced susceptibility to, and transmission of, HIV is part of the "Advances in HIV/AIDS research series" which endeavours to form a bridge between the research into HIV and acquired immunodeficiency syndrome (AIDS) and the practice of HIV/AIDS prevention, care and support in developing countries. Research findings have shown that being seropositive for HSV-2 can increase the risk of HIV acquisition among high-risk HIV-negative people exposed to HIV and, likewise, the infectiousness of individuals co-infected with HIV-1 and HSV-2 can increase during periods of HSV-2 reactivation. These observations have led to the initiation of several intervention trials and could ultimately lead to the setting of new priorities in public health and clinical practice. WHO has recently issued new guidelines for the syndromic management of genital ulcer disease that include antiviral treatment for lesions consistent with genital herpes. The United States Centers for Disease Control and Prevention issued updated Sexually Transmitted Diseases Treatment Guidelines in 2002 that recommended the use of type-specific serological tests for diagnosing HSV-2. Recently launched proof-of-concept, HSV-2 intervention trials in several countries will help to determine the proportion of new HIV infections that could be prevented by suppression of HSV-2, and the findings from these studies will inform those involved in setting prevention and treatment priorities and strategies in developing countries. PMID- 15356939 TI - Effectiveness of condoms in preventing sexually transmitted infections. AB - In June 2000, the United States National Institutes of Health (NIH) organized a review of the scientific evidence on the effectiveness of condoms in preventing sexually transmitted infections (STIs). The review concluded that condoms were effective in protecting against transmission of HIV to women and men and in reducing the risk of men becoming infected with gonorrhoea. Evidence for the effectiveness of condoms in preventing other STIs was considered to be insufficient. We review the findings of prospective studies published after June 2000 that evaluated the effectiveness of condoms in preventing STIs. We searched Medline for publications in English and included other articles, reports, and abstracts of which we were aware. These prospective studies, published since June 2000, show that condom use is associated with statistically significant protection of men and women against several other types of STIs, including chlamydial infection, gonorrhoea, herpes simplex virus type 2, and syphilis. Condoms may also be associated with protecting women against trichomoniasis. While no published prospective study has found protection against genital human papillomavirus (HPV) infection, two studies reported that condom use was associated with higher rates of regression of cervical intraepithelial neoplasia and clearance of cervical HPV infection in women and with regression of HPV associated penile lesions in men. Research findings available since the NIH review add considerably to the evidence of the effectiveness of condoms against STIs. Although condoms are not 100% effective, partial protection can substantially reduce the spread of STIs within populations. PMID- 15356940 TI - The natural history of HIV-1 and HIV-2 infections in adults in Africa: a literature review. AB - About 30 million people in Africa are estimated to be living with human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS), yet data about the natural history of infection on the continent are sparse. We reviewed the literature on the natural history of HIV-1 and HIV-2 infections among African adults. Only one study, conducted in rural Uganda, has reported on survival from the time of HIV-1 seroconversion: the median was 9.8 years, which is similar to that reported in developed countries in the early stages of the epidemic and consistent with the findings from the follow-up of individuals identified by serological testing during community-based prevalence studies from Africa. Progression to symptomatic disease was faster in Uganda than in developed countries, due largely to the high background level of morbidity. Various studies suggest that people infected with HIV-2 survive longer and the course of the disease is possibly more variable than in people infected with HIV-1. However no studies have investigated survival from time of seroconversion among people infected with HIV-2. The majority of patients in hospital in Africa with either HIV-1 or HIV-2 have the clinical features of AIDS just before they die, and many are severely immunosuppressed. This is similar to the situation in developed countries before the introduction of highly active antiretroviral therapy (HAART). Potentially preventable infections are the leading causes of death among individuals infected with HIV-1. Prophylactic regimens and better treatments could have some effect on survival, but major improvements in life expectancy will require HAART. PMID- 15356941 TI - SARS outbreak over, but concerns for lab safety remain. PMID- 15356942 TI - Cardiovascular disease--a global health time bomb. PMID- 15356943 TI - Brazilian genomics breakthrough offers hope for leptospirosis control. PMID- 15356944 TI - Rwandan genocide survivors in need of HIV treatment. PMID- 15356945 TI - New international convention allows use of DDT for malaria control. PMID- 15356946 TI - WHO's HIV/AIDS strategy under the spotlight. PMID- 15356947 TI - HIV treatment will boost prevention and strengthen health systems, says the World Health Report 2004. PMID- 15356948 TI - World Health Assembly adopts landmark strategies on diet and physical activity, and reproductive health. PMID- 15356949 TI - Human behaviour contributes to emergence of zoonoses. PMID- 15356950 TI - Images of healing. PMID- 15356951 TI - The idea of imagination: an analysis of "imagery". AB - BACKGROUND: Although the benefits of guided imagery are now more widely recognized, the definitions of imagery used by various health science disciplines are inconsistent. PURPOSE: To conduct an analysis of the concept of imagery as understood among the health science professions. METHODS: An inductive, descriptive means of inquiry was used to clarify the concept of imagery. Data were analyzed by categories: surrogates, attributes, contextual information (antecedents; consequences), related terms, and references. FINDINGS: The prevailing surrogate term for mental imagery is visualization. Attributes of imagery include psychophysiological, mental, quasi-reality, dynamic, and process. References include therapeutic settings. CONCLUSIONS: This concept analysis has led to the following working definition: "Imagery, a mental function, is a lived experience that is a dynamic, quasi-real, psychophysiological process." The analysis was intended to identify consensus on the concept of imagery within the health professions, and provide a foundation for further research. PMID- 15356952 TI - Stress-induced alternative gene splicing in mind-body medicine. AB - Recent research documents how psychosocial stress can alter the expression of the acetylcholinesterase gene to generate at least 3 alternative proteins that are implicated in a wide variety of normal mind-body functions, as well as pathologies. These range from early embryological development, plasticity of the brain in adulthood, post-traumatic stress disorder (PTSD), and stress-associated dysfunctions of the central nervous, endocrine, and immune systems, to age related neuropathologies. Such stress-induced alternative gene splicing is proposed here as a major mind-body pathway of psychosocial genomics-the modulation of gene expression by creative psychological, social, and cultural processes. We explore the types of research that are now needed to investigate how stress-induced alternative splicing of the acetylcholinesterase gene may play a pivotal role in the deep psychobiology of psychotherapy, meditation, spiritual rituals, and the experiencing of positive humanistic values that have been associated with mind-body medicine, such as compassion, beneficence, serenity, forgiveness, and gratitude. PMID- 15356953 TI - Mindfulness meditation to reduce symptoms after organ transplant: a pilot study. AB - CONTEXT: Solid organ transplant patients require life-long immune suppression that can produce distressing side effects and complications. OBJECTIVE: To evaluate the potential of Mindfulness-Based Stress Reduction (MBSR) to reduce symptoms of depression, anxiety, and sleep disturbance and improve quality of life after solid organ transplantation. DESIGN: Longitudinal with evaluations at baseline, postcourse and 3-month follow-up. PARTICIPANTS AND SETTING: Kidney, lung, or pancreas transplant recipients (N=20), aged 35 to 59 years, living in the community. INTERVENTION: An MBSR class (2.5 hours weekly, for 8 weeks), modeled after the program of Jon Kabat-Zinn. Home practice (goal: 45 minutes, 5 days weekly) was monitored. MAIN OUTCOME MEASURES: Self-report scales for depression (CES-D), anxiety (STAI-Y1), and sleep dysfunction (PSQI). RESULTS: Nineteen participants completed the course. Findings suggest improvement from baseline symptom scores for depression (P=.006) and sleep (P=.011) at the completion of the MBSR program. At 3 months, improvement in sleep continued (P=.002), and a significant improvement in anxiety scores was seen (P=.043); scores for both symptoms demonstrated a linear trend and dose-response relationship with practice time. In contrast, depression scores showed a quadratic trend, and at 3 months were no longer different from baseline. A composite symptom measure was significantly improved at 3-month follow-up (P=.007). Global and health-related quality of life ratings were not improved. Effects of group support and instructor attention were not controlled, and sample size and follow-up time were limited. A randomized trial to overcome these shortcomings should be done, as symptom distress in transplant recipients appears responsive to MBSR. PMID- 15356954 TI - Palliative care service at the NIH includes Reiki and other mind-body modalities. PMID- 15356955 TI - Genesis and evolution of the first biochemistry. PMID- 15356956 TI - Alcohol and thermally oxidized pufa induced oxidative stress: role of N-acetyl cysteine. AB - Alcohol related disabilities are one of the world's major public health concerns. The effects of alcohol intake include alteration of redox state, acetaldehyde and free radical production, which lead to membrane damage. The damage caused by alcohol is enhanced by polyunsaturated fatty acid ingestion. When alcohol is taken along with thermally oxidized sunflower oil, the toxicity is still more pronounced due to toxic metabolites produced during heating. In our study, we have analysed the effects of a thiol supplier N-acetyl cysteine on alcohol, thermally oxidized sunflower oil and alcohol + thermally oxidized sunflower oil induced toxic effects in male Wistar rats. The activities of liver marker enzymes (alkaline phosphatase and gamma-glutamyl transferase), triglycerides in plasma and lipid peroxidative indices (thiobarbituric acid reactive substances and hydroperoxides) were increased in these groups when compared to normal, which were brought down in N-acetyl cysteine treated groups. The antioxidant status (Superoxide dismutase, catalase, reduced glutathione, glutathione peroxidase) was decreased in tissues of these groups, which were found to be improved in N-acetyl cysteine treated groups. Thus our results show that N-acetyl cysteine regresses the oxidative damage induced by Alcohol, thermally oxidized sunflower oil and alcohol + thermally oxidized sunflower oil. PMID- 15356957 TI - Entropy calculator: getting the best from your multiple protein alignments. AB - Amino acid sequence alignment is an extremely useful tool in protein family analysis. Most family characteristics, such as the localization of functional residues, structural constraints and evolutionary relationships may be retrieved through the observation of the conservation pattern highlighted by the alignments. A quantitative score for the conservation in the alignment allows different stages of an alignment to be compared and consequently the alignment information to be efficiently exploited. Many scoring methods have been proposed during the last three decades. Claude Shannon's theory of communication (1948) paved the way for a consistent scoring of protein alignments by considering the residue (or symbol) frequency. A number of modifications have been proposed since that time, but the core statistical approach is still considered one of the best. By combining many database managing tools for treatment of protein sequences, a ClustalW software integration, a flexible symbols treatment and gap normalization functions, Entropy Calculator software has been developed. This new tool provides a global and optimal approach to multiple sequence alignment scoring by offering an easy graphic interface and a series of modification options that help in interpreting alignments and allow conservation pattern inferences to be performed. PMID- 15356958 TI - Influence of monosodium glutamate on circadian rhythms of lipid peroxidation products and antioxidants in rats. AB - Monosodium glutamate (MSG) was administrated subcutaneously for 60 days to Wistar rats and 24h rhythms of thiobarbituric acid reactive substances (TBARS) and antioxidants such as reduced glutathione, superoxide dismutase and catalase were studied. MSG treatment was found to advance TBARS and to delay the acrophases of GSH and catalase. Amplitude and mesor values of these rhythms were found to be altered during MSG treatment. As glutamate levels in brain were found to be significantly increased (in MSG), we hypothesize that increased glutamate levels in brain could alter these biochemical rhythms probaly by modulating the transmission in several areas/nuclei in brain. PMID- 15356959 TI - 4-hydroxy-2,3 trans-nonenal nuclear concentration inversely correlates with the growth rate of T leukemia cells. AB - 4-hydroxy-2,3 trans-nonenal is the major diffusible product generated by linoleic and arachidonic acids peroxidation. Its endogenous content is inversely related to the rate of cell proliferation and directly related to the level of cell differentiation. As previously reported, the nuclear localization of the aldehyde has been observed by means of a fluorescent antibody and confocal microscopy, and its concentration measured by electrospray/mass spectrometry, a sensitive and selective method for 4-hydroxy-2,3 trans-nonenal determination, on nuclear extracts of leukemic cells. With the aim to establish a possible correlation between the peroxidation product nuclear concentration and cell growth rate, Jurkat 6 leukemic cells have been used and the aldehyde measured by electrospray/mass spectrometry. The cells arrested in G1 show a content of 4 hydroxy-2,3 trans-nonenal significantly increased with respect to control ones. PMID- 15356960 TI - Mutations in respiratory chain complexes and human diseases. AB - Literary evidence for a link between mutations in genes encoding respiratory chain components and human disorders is reviewed with particular emphasis on defects in respiratory complexes III and IV and their assembly factors. To date, mutations in genes encoding cytochrome band QP-C structural subunits of cytochrome bc1 complex; the BCS1L assembly factor for the bc1 complex; structural subunits I-III of cytochrome c oxidase; as well as the SURF-1, COX10, SCO1, and SCO2 assembly factors for cytochrome c oxidase, have been reported. These mutations are responsible for different neuromuscular and non-neuromuscular human diseases. PMID- 15356961 TI - The reaction mechanism of plant peroxidases. AB - The catalysis of class III plant peroxidases is described based on the reaction scheme of horseradish peroxidase. The mechanism consists in four distinct steps: (a) binding of peroxide to the heme-Fe(III) to form a very unstable peroxide complex, Compound 0; (b) oxidation of the iron to generate Compound I, a ferryl species with a pi-cation radical in the porphyrin ring; (c) reduction of Compound I by one substrate molecule to produce a substrate radical and another ferryl species, Compound II; (d) reduction of Compound II by a second substrate molecute to release a second substrate radical and regenerate the native enzyme. Under unfavourable conditions some inactive enzyme species can be formed, known as dead end species. Two calcium ions are normally found in plant peroxidases and appear to be important for the catalytic efficiency. PMID- 15356963 TI - The role of N-acetylcysteine in preventing radiographic contrast-induced nephropathy. AB - INTRODUCTION: There have been many small randomized controlled trials evaluating the effectiveness of N-acetylcysteine (NAC) in preventing radiographic contrast induced nephropathy. Most studies have suggested a beneficial NAC effect. This meta-analysis describes the effect of NAC in the prevention of radiographic contrast-induced nephropathy in the aggregated trial data. METHODS: A search using MEDLINE from 1966 to December 2003 identified all randomized control trials that evaluated NAC in those patients at risk of acute renal failure (ARF) following either angiographic or CT scan contrast exposure. All studies included in the review employed the use of either low-osmolar (n = 9 trials) or iso osmolar (n = 2 trials) contrast agents. The outcome of interest was ARF as defined by a rise in serum creatinine (Cr > or = 0.5 mg/dl rise or > 25% increase from baseline) after exposure to contrast. The data were aggregated by the methods of Mantel and Haenszel. RESULTS: The overall summary odds ratio estimate of 0.46 (95% confidence interval 0.32 - 0.66) suggests a strong protective effect of NAC in preventing radiographic-induced nephropathy. CONCLUSION: In summary, there is good aggregate trial evidence to suggest that patients who have an elevated serum creatinine level at baseline benefit from receiving periprocedure NAC in the prevention of contrast-induced ARF. PMID- 15356962 TI - Roles for holes: are cavities in proteins mere packing defects? AB - Atomic packing in proteins is not optimized, most structures containing internal cavities, which have been identified by molecular modelling and characterized experimentally. Cavities seem to play a role in assisting conformational changes between domains or subunit interfaces. Comparison between homologous proteins from thermophiles and mesophiles indicates that optimizing packing enhances stabilization at the expense of flexibility. For proteins which interact with small ligands or substrates, cavities seem to play a role in controlling binding and catalysis, rather than being mere "packing defects". We believe that a more complete analysis on the localization, conservation and role of cavities in protein structures (by modelling and site-directed mutagenesis), will reveal that rather than being randomly distributed, they are located in key positions to allow structural dynamics and thereby functional control. PMID- 15356964 TI - Plasma leptin concentration in patients with acute renal failure. AB - BACKGROUND: Acute renal failure (ARF) is characterized by impaired excretory, endocrine, homeostatic and metabolic function of the kidneys. It is well-known that leptin is an adipose-derived polypeptide hormone which is predominantly biodegraded by the kidneys. Therefore, plasma leptin concentration is increased in chronic renal failure (CRF). However, its' concentrations in patients with ARF were not investigated until now. The aim of the present study was to evaluate plasma leptin concentration in patients with ARF. PATIENTS AND METHODS: 27 patients with ARF (age 44 +/- 4 years, BMI 26.0 +/- 0.9 kg/m2, means +/- SEM, 17 patients 15 M, 2 F recovered kidney function and 10 patients 7 M, 3 F died during the anuric phase), 27 hemodialysis patients (22 M, 5 F; age 45 +/- 2 years; BMI 26.2 +/- 0.8 kg/m2) with chronic renal failure (CRF) and 27 healthy subjects (HS) (22 M, 5 F; age 42 +/- 3 years; BMI 25.9 +/- 0.6 kg/m2) were examined. In patients with ARF, blood samples for plasma leptin and routinely assessed biochemical parameters were withdrawn before the first HD session (I), and in patients who survived a second time 5 days later during the anuric/oliguric phase (II), and a third one during the polyuric phase before discharge of the patient from hospital (III). In patients with CRF all examined parameters were estimated only once before a subsequent HD session. RESULTS: Patients with ARF (before the first HD session) and CRF did not differ significantly with respect to BMI, serum creatinine and blood hydrogen ion concentrations. Plasma leptin level in patients with ARF before the first HD session was similar to values obtained in HS, but significantly lower (p < 0.01) than in patients with CRF (2.5 (1.9 - 8.2) vs. 3.4 (2.5 - 8.3) vs. 8.4 (2.9 - 16.9) ng/ml in ARF, HS and CRF, respectively). There was no significant difference in leptinemia between patients with ARF who survived and who died. In patients with ARF who survived, improvement ofrenal function was accompanied by a slightly (not significant) declining tendency in plasma leptin concentration (5.6 +/- 2.2 vs. 4.8 +/- 1.7 vs. 4.5 +/- 1.3 ng/ml; I, II, III phases of ARF, respectively). CONCLUSIONS: In contrast to hemodialysis patients with chronic renal failure, patients with acute renal failure are characterized by normal plasma leptin concentration. Thus, difference in leptinemia between patients with chronic and acute renal failure seems to be due to preservation of large amounts of active renal parenchyma in ARF patients. PMID- 15356965 TI - Estimation of glomerular filtration rate in obese patients with chronic renal impairment based on serum cystatin C levels. AB - BACKGROUND: Serum cystatin C (Scyst) has an obvious advantage in recognizing the initial stages of renal impairment. However, several recent studies suggest that Scyst may also be affected by some nonglomerular factors such as thyroid dysfunction, glucocorticoid administration or metabolic status of the diabetic patient. The aim of this study was to evaluate whether obesity could affect Scyst. PATIENTS AND METHODS: The study was performed in 33 patients (mean age 49.1 +/- 6.3 years) with chronic renal disease (Scr = 227 +/- 118 micromol/l) and BMI = 35.6 +/- 1.8 kg/m2, and in 78 patients (mean age 43.4 +/- 5.1 years) with chronic renal disease (Scr = 245 +/- 111 micromol/l) and BMI = 24.0 +/- 1.8 kg/m2. Glomerular filtration rate (GFR) was determined using renal inulin clearance (Cin) under conditions of stabilized plasma concentrations and water loading. Scyst was measured using immunonephelometry. For statistical evaluation, linear regression analysis and receiver-operating characteristic (ROC) curve analysis were used. RESULTS: A significant correlation (r = 0.956, p < 0.001) between l/Scyst and Cin was demonstrated in patients with BMI > or = 30 kg/m2 (Group A). Similarly, a significant correlation (r = 0.900, p < 0.001) between l/Scys and Cin was found in patients with BMI < 30 kg/m2 (Group B). There was no significant difference between the regression straight lines characterizing these relationships. ROC curve analysis (using a cut-off value for Cin = 30 ml/min/1.73 m2) did not show significant differences in AUC, sensitivity and specificity for Scyst between obese and nonobese patients. CONCLUSION: The results suggest that evaluation of GFR based on Scyst in obese patients need not differ from that in nonobese ones. PMID- 15356966 TI - A simple method for correcting overestimated glomerular filtration rate in obese subjects evaluated by the Cockcroft and Gault formula: a comparison with 51Cr EDTA clearance. AB - AIM: The Cockcroft and Gault formula is a quick and reliable method for calculating creatinine clearance without a 24-hour urine collection (CG-cl). In obese subjects an excess of fat mass provokes a reduction in daily creatinine urine excretion per body kilo weight and is responsible for overestimated renal function when calculated by CG-cl. The aim of this study was to devise a simple correction method which could also make use of CG-cl in obese subjects. PATIENTS AND METHODS: In 52 subjects with a body mass index (BMI) > 25, renal function was assessed by simultaneously determining creatinine clearance using 24-hour urine collection (Cr-cl) and the CG-cl. The percentage difference between the 2 clearances (delta %) was correlated with BMI for each patient using simple linear regression analysis. The estimated regression model (delta% = 1.217 BMI-- 24.81) provided the following CG-cl correction formula for obese subjects: Corrected CG cl = CG-cl (1.25 - 0.012 BMI). Its validity was evaluated in another group of 20 subjects with BMI > 25 by comparing the results obtained with Corrected CG-cl to those obtained by CG-cl and MDRD formula (MDRD-cl) using the clearance of 51Cr EDTA (5 Cr-EDTA-cl) as the GFR measurement gold standard. RESULTS AND CONCLUSION: Linear regression analysis of CG-cl, MDRD-cl and Corrected CG-cl compared to 5tCr EDTA-cl (considered as the independent variable) resulted in the following determination coefficients (R2): 0.687; 0.818; 0.947, respectively. In conclusion, this formula can be considered a quick and reliable method for CG-cl correction in obese subjects. PMID- 15356967 TI - Long-term comparison of a calcium-free phosphate binder and calcium carbonate- phosphorus metabolism and cardiovascular calcification. AB - BACKGROUND: Calcium carbonate used as a phosphate binder may contribute to cardiovascular calcification. Long-term comparisons of sevelamer, a non-calcium polymeric phosphate binder, and calcium carbonate (CC) are lacking. METHODS: 114 adult hemodialysis patients were randomly assigned to open label sevelamer or CC for 52 weeks. Study efficacy endpoints included changes in serum phosphorus, calcium, calcium-phosphorus product, and lipids. In addition, initial and sequential electron beam computerized tomography scans were performed to assess cardiovascular calcification status and change during follow-up. Safety endpoints were serum biochemistry, blood cell counts and adverse events. RESULTS: Patients receiving sevelamer had a similar reduction in serum phosphorus as patients receiving CC (sevelamer -0.58 +/- 0.68 mmol/l, CC -0.52 +/- 0.50 mmol/l; p = 0.62). Reductions in calcium-phosphorus product were not significantly different (sevelamer -1.4 +/- 1.7 mmol2/l2, CC -0.9 +/- 1.2 mmol2/l2; p = 0.12). CC produced significantly more hypercalcemia (> 2.8 mmol/l in 0% sevelamer and 19% CC patients, p < 0.01) and suppressed intact parathyroid hormone below 150 pg/ml in the majority of patients. Sevelamer patients experienced significant (p < 0.01) reductions in total (-1.2 +/- 0.9 mmol/l, -24%) and LDL cholesterol (-1.2 +/- 0.9 mmol/l, -30%). CC patients had significant increases in coronary artery (median +34%, p < 0.01) and aortic calcification (median +32%, p < 0.01) that were not observed in sevelamer-treated patients. Patients on sevelamer required more grams of binder (sevelamer 5.9 g vs. CC 3.9 g) and experienced more dyspepsia than patients on calcium carbonate. CONCLUSIONS: Sevelamer is an effective phosphate binder that unlike calcium carbonate is not associated with progressive cardiovascular calcification in hemodialysis patients. PMID- 15356968 TI - Rapid, high-dose intravenous iron sucrose therapy in 2 Jehovah's Witness patients with severe anemia, iron deficiency and chronic kidney disease. AB - AIMS: Two patients with chronic kidney disease presented with severe anemia and iron deficiency. Because of their religious beliefs, red blood cell transfusions were not possible, and an aggressive therapeutic regimen of iron replenishment was instituted. MATERIAL AND METHODS: The regimen included epoetin, folic acid and high-dose intravenous iron sucrose infusions over multiple successive days (total dosages of 2 and 3.5 g). RESULTS: The patients' iron stores were replenished and an erythropoietic response ensued subsequent to this aggressive and unique therapeutic regimen. There were no side effects observed which could be attributed to iron sucrose, and both patients stabilized and were discharged after 3 - 4 weeks. CONCLUSION: In patients with chronic kidney disease who are severely anemic and iron-deficient and where transfusions are not possible, an aggressive regimen of multiple high-dose iron sucrose infusions may be both safe and effective. PMID- 15356969 TI - Glomerular monocyte/macrophage influx correlates strongly with complement activation in 1-week protocol kidney allograft biopsies. AB - BACKGROUND: The specific role of monocytes/macrophages (MO) in kidney graft rejection is not yet fully elucidated. In a recent protocol biopsy study of living-donor recipients, we demonstrated massive capillary influx of MO, associated with severe complement activation and acute rejection (AR) 1 week after transplantation [Sund et al.]. To gain further insight into the possible relationship between MO and complement activation, we analyzed glomerular and interstitial MO in these biopsies. METHODS: Twenty-seven protocol biopsies were stained with antibodies to calprotectin (L1) and CD68 as markers for MO. Cells were counted as an average number per glomerulus and as an average number per defined visual field in the interstitium. Polymorphonuclear leukocytes (PMN) were counted in glomeruli and interstitium by light microscopy. Baseline specimens from 10 of the patients served as controls. The results were compared with data on deposition of complement from the foregoing study, and with histopathologic and clinical data on AR. RESULTS: Cases with diffuse C4d deposition in peritubular capillaries consistent with acute antibody-mediated rejection (AbAR) (n = 4) had significantly higher numbers of intraglomerular MO than the other protocol biopsies (L1: median 20.7 vs 3.6, p = 0.0002; CD68: median 10.1 vs. 2.0, p = 0.0008). With a cut-off of 10 L1-positive and 6 CD68-positive MO, the specificity for the diagnosis of AbAR was 96% and 91%, respectively. The number of interstitial MO was significantly higher in patients with AR than in those without, but in contrast to glomerular MO, interstitial MO could not discriminate between complement positive and negative AR. The number of glomerular and interstitial PMNs was significantly higher in the AbAR group than in the other protocol biopsies. CONCLUSIONS: The strong correlation between complement activation and early glomerular influx of MO in the kidney allograft suggests a causal relationship between these 2 events. At 1 week after transplantation, a number of 10 L1-positive and 6 CD68-positive MO per glomerulus indicates AbAR. PMID- 15356970 TI - Normalization of oxidative stress parameters after kidney transplant is secondary to full recovery of renal function. AB - BACKGROUND: It is well-known that hemodialysis patients experience increased oxidative stress, which is believed to cause numerous uremia-related complications. Retention of water-soluble toxins as well as protein-bound toxins is due to renal failure. Kidney transplantation restores, at least partially, the fundamental processes of glomerular filtration which eliminates toxic solutes. The aim of this study was to determine the levels of several different glycoxydative stress-related parameters after kidney transplantation. PATIENTS AND METHODS: A cross-sectional study was carried out on 30 subjects: 10 kidney transplanted patients with chronic renal failure (Tx-CRF), 10 kidney-transplanted patients with normal renal function (Tx-N) and 10 controls (Ctr). The groups were comparable with respect to age and gender. The following glycoxydative stress markers were determined by HPLC analysis: albumin-bound and free pentosidine, low molecular weight-advanced glycation end products (LMW-AGEs), advanced oxidation protein products (AOPP) and low-molecular weight carbonyls (LMW-C). The total antioxidant serum capacity was monitored by measuring both the ferric reducing/antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC). RESULTS: With respect to the controls, the Tx-CRF patients had higher levels of pentosidine (2.66 +/- 0.98 vs 1.45 +/- 1.1 pmol/mg), LMW-AGE (47.55 +/- 39.74 vs 15.45 +/- 6.39 a.u./ml), and AOPP (6.71 +/- 0.78 vs 4.81 +/- 0.32 a.u./mg) while Tx patients with normal kidney function had levels of these compounds that were comparable to the controls, except for the LMW-AGEs which were higher. Levels of LMW-AGEs, pentosidine, LMW-C and AOPP were inversely correlated to creatinine clearance. The total antioxidation serum capacity was paradoxically higher in Tx patients than in the controls, regardless of kidney function. FRAP as well as ORAC, were correlated to uric acid (r = 0.62, p < 0.001; r = 0.54, p < 0.01). CONCLUSIONS: The reported data indicate that kidney transplantation seems to restore a nearly normal level of glycoxidative stress markers, but a complete remission is only possible when the renal function is normal. An increase of total antioxidant power of serum in transplanted patients was reported, as probable effect of uric acid high levels. PMID- 15356971 TI - IgA-dominant glomerulonephritis associated with hepatitis A. AB - Unlike hepatitis B and C, renal involvement has been extremely uncommon in patients with hepatitis Avirus (HAV) infection. Nephrotic syndrome has been documented as a rare complication in association with HAV infection. In this report, we describe a patient with serologically documented HAV infection, who presented with nephrotic syndrome. The renal biopsy showed an immunoglobulin A- (IgA) dominant glomerulonephritis (GN) with subendothelial immune deposits. This is the second biopsy-proven case report of a patient with acute HAV associated with IgA-dominant immune complex glomerulonephritis and nephrotic syndrome. This is perhaps the first case in which a patient experienced both IgA-dominant glomerulonephritis and cutaneous cryoglobulinemic vasculitis. PMID- 15356972 TI - Vasculitic purpura with antineutrophil cytoplasmic antibody-positive acute renal failure in a patient with Streptococcus bovis case and Neisseria subflava bacteremia and subacute endocarditis. AB - Subacute bacterial endocarditis is frequently associated with extracardiac manifestations and renal failure. Clinical variety of endocarditis manifestation is wide and has the potential to mimic vasculitis. Whereas Streptococcus bovis is often isolated and associated with colonic tumors, Neisseriaceae are rarely found. An association of subacute bacterial endocarditis and antineutrophil cytoplasmic antibodies has been described. We report on a 62-year-old man who was admitted to our hospital with acute oliguric renal failure and a nonpruritic purpural rush without fever. Antineutrophil cytoplasmic antibody diagnostic revealed perinuclear staining with a titre of 1 : 512 and antiproteinase-3 specificity. Immune complex-mediated glomerulonephritis without extracapillary proliferation was diagnosed in renal biopsy. Finally, blood cultures became positive for Streptococcus bovis and Neisseria flava. Echocardiography showed mobile vegetations on tricuspid valve. Under treatment with penicillin G and gentamicin, skin efflorescences and renal function recovered, but vegetations increased. A colonic tumor could be excluded, a disastrous dental status may have been a predisposal factor. When classical findings of subacute bacterial endocarditis are less clear, the presence of renal failure and antineutrophil cytoplasmic antibodies in absence of fever may lead to misdiagnosis and deleterious immunosuppressive therapy. Neisseria subflava, an upper respiratory tract commensal, may cause subacute bacterial endocarditis without typical symptoms. PMID- 15356973 TI - Progressive nephropathy associated with mitochondrial tRNA gene mutation. AB - Mitochondrial DNA plays a crucial role in oxidative production of energy. Thus, defects in mitochondrial DNA can affect virtually all organ systems. The point mutation A --> G at position 3243 in the mitochondrial tRNAleu(UUR) gene is the cause of several distinct types of mitochondrial cytopathy and several clinical phenotypes, including encephalomyopathy with lactic acidosis and stroke-like episodes and maternally inherited diabetes and deafness. This mutation has been recently described also in association with kidney disease, mainly focal and segmental glomerulosclerosis. At present, little is known about the prevalence of this mitochondrial nephropathy, its clinical course and the pathogenesis of glomerular damage. We describe 2 unrelated patients, who presented with proteinuria and progressed to end-stage renal failure. Other clinical features were short stature, severe headache, hearing loss, diabetes mellitus and hypertrophic cardiomyopathy. The main histological finding was an increased number of abnormal mitochondria in tubular cells and podocytes. Analysis of mitochondrial DNA from leukocytes and urine sediment revealed heteroplasmy for the A3243G mutation in tRNAleu(UUR) gene in both patients. Recognition of the characteristic clinical and histological features of the mitochondrial A3243G mutation-associated glomerulopathy will enable correct diagnosis and better management of a disease which is likely to be underdiagnosed. PMID- 15356974 TI - Late-onset primary hyperoxaluria triggered by hypothyroidism and presenting as rapidly progressive renal failure--description of a new mutation. AB - Primary hyperoxaluria type 1 (PH1) is a rare autosomal metabolic recessive disease, caused by the deficiency of the liver peroxysomal alanine:glyoxylate aminotransferase (AGT), characterized by accumulation of calcium oxalate crystals in kidneys and others organs. We present the case of an elderly woman with PH1, presenting as acute renal failure. Precipitation of calcium oxalate crystals was probably due to amiodarone-induced severe hypothyroidism. Residual AGT activity is associated with the G170R (G630A) mutation. A new mutation of AGT, called R36C, was also discovered; the role of this new mutation is actually not known. PMID- 15356975 TI - A rare cause of anemia due to intestinal tuberculosis in a renal transplant recipient. AB - A renal transplant recipient with stable allograft function presented with massive hemorrhagic diarrhea and severe anemia. No microbial infection could be found in stool cultures. Early colonoscopy showed severe colitis with ulceration. Histological samples confirmed granulomatous inflammation with acid-resistant Ziehl-Neelson-positive microorganisms of mycobacterial type. Polymerase chain reaction (PCR) analysis of native mucosal biopsies specified the infectious organism as Mycobacterium tuberculosis complex. The patient responded well to antimycobacterial therapy and was still asymptomatic after 6 months with a stable graft function. Our case shows that tuberculosis can be a severe clinical problem in transplant recipients. Most of the patients with intestinal tuberculosis, reported to literature, were diagnosed post mortem or after explorative laparotomy and bowel resection. Thus, intestinal tuberculosis should be considered when a transplant recipient shows abdominal symptoms with no clear evidence of another infection. Proper diagnosis and treatment resulted in a beneficial outcome in our patient. PMID- 15356976 TI - Indwelled femoral vein non-cuffed, double-lumen hemodialysis catheter complicated by pulmonary thromboembolism. AB - Non-cuffed, double-lumen hemodialysis (HD) catheters can be inserted at the bedside in the femoral, internal jugular or subclavian position. The femoral route is less risky, and the incidence of life-threatening complications is lower for femoral cannulation than for internal jugular and subclavian cannulations. However, here we describe a life-threatening complication of an extensive deep vein thrombosis and subsequent pulmonary thromboembolism following femoral cannulation of a double-lumen HD catheter. The possible mechanisms and treatment for this potentially fatal thromboembolic event are discussed in this report. PMID- 15356977 TI - Mizoribine pulse therapy for patients with flares of lupus nephritis: a 1-year observation. PMID- 15356978 TI - Making IT ends meet. Susquehanna Health System outsources Siemens' prized product to nearby hospitals looking for an affordable alternative. AB - Susquehanna Health System has been offering small, rural hospitals a more affordable information technology system through a unique outsourcing agreement. The system's CIO, Pamela Wirth, left, has advocated the provider-to-provider arrangement, which could draw the ire of some vendors. PMID- 15356979 TI - Cold calls, hot deals. Broadlane's phone pitch: no membership required. PMID- 15356980 TI - Creating a coalition. Cafe Rx to tackle woes of E-prescription movement. PMID- 15356981 TI - Golden years. Hospitals redesigning retirement plans to help get handle on costs. PMID- 15356982 TI - Tax-exempt targets. IRS introduces initiative to examine executive compensation, excess benefits. PMID- 15356983 TI - Change is not so bad. Host of outpatient PPS adjustments draws little reaction from hospital industry. PMID- 15356984 TI - Going retro. Health plans try to control costs by bringing back some old practices. PMID- 15356985 TI - Piece of the pie. Community health centers unhappy with their slice. PMID- 15356986 TI - State of alert. Calif nursing homes battle over matching-funds tax. PMID- 15356987 TI - Hanging tough. Modern Healthcare's group purchasing survey reveals that the GPO industry continues to grow despite a stagnant customer pool. PMID- 15356988 TI - A not-so-free choice. Hospitals, AHA must decide whether to fight or settle uninsured billing cases. PMID- 15356989 TI - The promise and the reality. The sky's the limit on healthcare IT, but first we have to get off the ground. PMID- 15356990 TI - By the numbers. Top 20 pharmaceutical companies. Ranked by U.S. sales, April 2003 through March 2004. PMID- 15356991 TI - This bud's for the U.S. Canada's relaxed drug laws may be fueling a boom in marijuana exports to America. PMID- 15356993 TI - Why lines must be drawn. Stem cells present a complex moral issue. Shame on Democrats for polarizing it. PMID- 15356992 TI - Stress and the superdad. Like the supermoms before them, today's fathers are struggling to balance work and home. PMID- 15356994 TI - Suppression of protein kinase C and nuclear oncogene expression as possible action mechanisms of cancer chemoprevention by Curcumin. AB - Curcumin (diferuloylmethane) is a major naturally-occurring polyphenol of Curcuma species, which is commonly used as a yellow coloring and flavoring agent in foods. Curcumin has shown anti-carcinogenic activity in animal models. Curcumin possesses anti-inflammatory activity and is a potent inhibitor of reactive oxygen generating enzymes such as lipoxygenase/cyclooxygenase, xanthine dehydrogenase/oxidase and inducible nitric oxide synthase; and an effective inducer of heme oxygenase-1. Curcumin is also a potent inhibitor of protein kinase C (PKC), EGF(Epidermal growth factor)-receptor tyrosine kinase and IkappaB kinase. Subsequently, curcumin inhibits the activation of NF(nucleor factor)kappaB and the expressions of oncogenes including c-jun, c-fos, c-myc, NIK, MAPKs, ERK, ELK, PI3K, Akt, CDKs and iNOS. It is proposed that curcumin may suppress tumor promotion through blocking signal transduction pathways in the target cells. The oxidant tumor promoter TPA activates PKC by reacting with zinc thiolates present within the regulatory domain, while the oxidized form of cancer chemopreventive agent such as curcumin can inactivate PKC by oxidizing the vicinal thiols present within the catalytic domain. Recent studies indicated that proteasome-mediated degradation of cell proteins play a pivotal role in the regulation of several basic cellular processes including differentiation, proliferation, cell cycling, and apoptosis. It has been demonstrated that curcumin-induced apoptosis is mediated through the impairment of ubiquitin proteasome pathway. Curcumin was first biotransformed to dihydrocurcumin and tetrahydrocurcumin and that these compounds subsequently were converted to monoglucuronide conjugates. These results suggest that curcumin-glucuronide, dihydrocurcumin-glucuronide, tetrahydrocurcumin-glucuronide and tetrahydrocurcumin are the major metabolites of curcumin in mice, rats and humans. PMID- 15356995 TI - Pharmacokinetic changes in drugs during protein-calorie malnutrition: correlation between drug metabolism and hepatic microsomal cytochrome P450 isozymes. AB - The rats with protein-calorie malnutrition (PCM, 5% casein diet for a period of 4 week) were reported to exhibit 60 and 80% suppression in the hepatic microsomal cytochrome P450 (CYP) 1A2 and CYP2C11 levels, respectively, and 40-50% decreases in CYP2E1 and CYP3A1/2 levels compared to control (23% casein diet for a period of 4-week) based on Western blot analysis. In addition, Northern blot analysis showed that CYP1A2, CYP2E1, CYP2C11, and CYP3A1/2 mRNAs decreased in the state of PCM as well. Hence, pharmacokinetic changes of the drugs in rats with PCM [especially the area under the plasma concentration-time curve from time zero to time infinity (AUC) changes of metabolite(s)] reported from literatures were tried to explain in terms of CYP isozyme changes in the rats. Otherwise, the time averaged nonrenal clearance (CL NR) of parent drug was compared. Pharmacokinetic changes of the drugs in other types of malnutritional state, such as kwashiorkor and marasmus, in both human and animal models were also compared. The drugs reviewed are as follows: diuretics, antibiotics, anticancer agents, antiepileptics, antiarrythmics, analgesics, xanthines, antimalarials, and miscellaneous. PMID- 15356996 TI - Synthesis and antitubercular activity of 6-chloro (unsubstituted)- 2-methoxy-9 substituted acridine derivatives. AB - Several analogues of the general formulae 2-methoxy-9-substituted acridine and 6 chloro-2-methoxy-9-substituted acridine were synthesized and evaluated in vitro at 6.25 microg/mL against M. tuberculosis H37Rv. Compounds 15 and 17 showed potential antitubercular activity with 100% inhibition to the virulent mycobacterium. PMID- 15356997 TI - Spectrophotometric determination of nizatidine and ranitidine through charge transfer complex formation. AB - Two Spectrophotometric procedures are presented for the determination of two commonly used H2-receptor antagonists, nizatidine (I) and ranitidine hydrochloride (II). The methods are based mainly on charge transfer complexation reaction of these drugs with either p-chloranilic acid (rho-CA) or 2, 3 dichloro 5, 6-dicyanoquinone (DDQ). The produced colored products are quantified spectrophotometrically at 515 and 467 nm in chloranilic acid and DDQ methods, respectively. The molar ratios for the reaction products and the optimum assay conditions were studied. The methods determine the cited drugs in concentration ranges of 20-200 and 20-160 microg/mL for nizatidine and ranges of 20-240 and 20 140 microg/mL for ranitidine with chloranilic acid and DDQ methods, respectively. A more detailed investigation of the complexes formed was made with respect to their composition, association constant, molar absorptivity and free energy change. The proposed procedures were successfully utilized in the determination of the drugs in pharmaceutical preparations. The standard addition method was applied by adding nizatidine and ranitidine to the previously analyzed tablets or capsules. The recovery of each drug was calculated by comparing the concentration obtained from the spiked mixtures with those of the pure drug. The results of analysis of commercial tablets and the recovery study (standard addition method) of the cited drugs suggested that there is no interference from any excipients, which are present in tablets or capsules. Statistical comparison of the results was performed with regard to accuracy and precision using student's t-test and F ratio at 95% confidence level. There is no significant difference between the reported and proposed methods with regard to accuracy and precision. PMID- 15356998 TI - A new cycloartane glycoside from Camptosorus sibiricus Rupr. AB - A new cycloartane glycoside was isolated from the dried whole herbs of Camptosorus sibiricus Rupr By means of chemical (hydrolysis) and spectroscopic methods (IR, 1D, and 2D NMR, ESI-MS), the structure was established as 3beta, 7beta, 24beta, 25, 30-pentahydroxycycloartane-3-omicron-beta-D-glucopyranoside-24 omicron-beta-D-glucopyranoside (1). PMID- 15356999 TI - Triterpenoid saponin from Viola hondoensis W. Becker et H Boss. and their effect on MMP-1 and type I procollagen expression. AB - Bioassay-guided fractionation has led to the isolation of triterpenoid saponins such as Acutoside A (3-omicron-[omicron-beta-D-glucopyranosyl-(1 --> 2)-omicron beta-D-glucopyranosyl] oleanolic acid) from the whole plants of Viola hondoensis. Among them, Saponin 1 exhibited potent inhibitory activity against matrix metalloproteinase (MMP)-1, and prevented the UV-induced changes in the MMP-1 expression. In addition, compound was isolated from this plant for the first time. PMID- 15357000 TI - Cytotoxic constituents of Diadema setosum. AB - 5,8-Epidioxycholest-6-en-3-ol (1), cholesterol (2), glycerol 1-palmitate (3) and glycerol 1,3-dioleate-2-stearate (4) were isolated from the methanol extract of the sea urchin Diadema setosum, which was collected from the Halong sea, Vietnam. Chemical structures were established based on extensive 1D, 2D-NMR, FAB-MS, El-MS spectroscopic data and GC-MS analysis. The NMR spectral data of compound 1 were reassigned by using HMQC and HMBC. Compound 1 was found to have strong cytotoxic effect against various cancer cell lines, such as KB (IC50, 2.0 microg/mL), FL (ICso, 3.93 microg/mL), and Hep-2 (IC50, 2.4 microg/mL) by in vitro assay. PMID- 15357001 TI - Inhibitory lignans against NFAT transcription factor from Acanthopanax koreanum. AB - Three lignans isolated from the roots of A. koreanum (Araliaceae), namely eleutheroside E (1), tortoside A (2), and hemiariensin (4), were evaluated for their ability to inhibit NFAT transcription factor. Of these compounds, compound 4, possessing a diarylbutane skeleton, exhibited potent inhibitory activity against NFAT transcription factor (IC50: 36.3 +/- 2.5 microM). However, the activities of 1 (IC50: > 500 microM) and 2 (IC50: 136.1 +/- 9.4 microM), which possess bisaryldioxabicyclooctane skeletons, were lower. As the lignan derivatives of the same skeletons, hinokinin (5) and (-)-yatein (6) with diarylbutane skeletons and (+)-syringaresinol (3) with a bisaryldioxabicyclooctane skeleton were also studied for their inhibitory effects on NFAT transcription factor. PMID- 15357002 TI - A sphingolipid and tyrosinase inhibitors from the fruiting body of Phellinus linteus. AB - This paper for the first time reports the isolation of 5 compounds from Phellinus linteus. A sphingolipid (1) and two tyrosinase inhibitory compounds (2, 3) along with two carboxylic acids (4, 5), were isolated from the fruiting body of Phellinus linteus (Berk & Curt) Aoshima. The structure of compound 1 was identified as 1-omicron-beta-D-glucopyranosyl-(2S, 3R, 4E, 8E)-2-[(2R)-2 hydroxyhexadecanoylamino]-9-methyl-4,8-octadecadiene-1,3-diol, known as cerebroside B, based on spectroscopic methods such as 1 D and 2D NMR as well as by acid hydrolysis. Compounds 2 -5 were identified as protocatechualdehyde (2), 5 hydroxymethyl-2-furaldehyde (HMF) (3), succinic acid (4), and fumaric acid (5) based on the spectroscopic evidence. Compounds 2 and 3 inhibited the oxidation of L-tyrosine catalyzed by mushroom tyrosinase with an IC50 of 0.40 and 90.8 microg/mL, respectively. The inhibitory kinetics, which were analyzed by the Lineweaver-Burk plots, were found to be competitive and noncompetitive inhibitors with a Ki of 1.1 microM and 1.4 mM, respectively. PMID- 15357003 TI - Modulation of suppressive activity of lipopolysaccharide-induced nitric oxide production by glycosidation of flavonoids. AB - Flavonoids have been demonstrated to exhibit a wide range of biological activities including anti-inflammatory and neuroprotective actions. Although a significant amount of flavonoids has been identified to be present as glycosides in medicinal plants, determinations of the biological activities of flavonoids were mainly carried out with aglycones of flavonoids. Therefore, the exact role of the glycosidation of flavonoid aglycones needs to be established. In an attempt to understand the possible role of glycosidation on the modulation of the biological activities of flavonoids, diverse glycosides of kaempferol, quercetin, and aromadendrin were examined in terms of their anti-inflammatory activity determined with the suppression of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in BV2 microglial cells. The results indicated that glycosidation of aglycones attenuated the suppressive activity of aglycones on LPS-induced NO production. Although attenuated, some of glycosides, depending on the position and degree of glycosidation, maintained the inhibitory capability of LPS-induced NO production. These findings suggest that glycosidation of flavonoid aglycones should be considered as an important modulator of the biological activities of flavonoids. PMID- 15357004 TI - PKC downstream of Pl3-kinase regulates peroxynitrite formation for Nrf2-mediated GSTA2 induction. AB - The protective adaptive response to electrophiles and reactive oxygen species is mediated by the induction of phase II detoxifying genes including glutathione S transferases (GSTs). NF-E2-related factor-2 (Nrf2) phosphorylation by protein kinase C (PKC) is a critical event for its nuclear translocation in response to oxidative stress. Previously, we have shown that peroxynitrite plays a role in activation of Nrf2 and Nrf2 binding to the antioxidant response element (ARE) via the pathway of phosphatidylinositol 3-kinase (Pl3-kinase) and that nitric oxide synthase in hepatocytes is required for GSTA2 induction. In view of the importance of PKC and Pl3-kinase in Nrf2-mediated GST induction, we investigated the role of these kinases in peroxynitrite formation for GSTA2 induction by oxidative stress and determined the relationship between PKC and Pl3-kinase. Although PKC activation by phorbol 12-myristate-13-acetate (PMA) did not increase the extents of constitutive and inducible GSTA2 expression, either PKC depletion by PMA or PKC inhibition by staurosporine significantly inhibited GSTA2 induction by tert-butylhydroquinone (t-BHQ) a prooxidant chemical. Therefore, the basal PKC activity is requisite for GSTA2 induction. 3-Morpholinosydnonimine (SIN-1), which decomposes and yields peroxynitrite, induced GSTA2, which was not inhibited by PKC depletion, but slightly enhanced by PKC activation, suggesting that PKC promotes peroxynitrite formation for Nrf2-mediated GSTA2 induction. Treatment of cells with S-nitroso-N-acetyl-penicillamine (SNAP), an exogenous NO donor, in combination with t-BHQ may produce peroxynitrite. GSTA2 induction by SNAP + t-BHQ was not decreased by PKC depletion, but rather enhanced by PKC activation, showing that the activity of PKC might be required for peroxynitrite formation. LY294002 a Pl3-kinase inhibitor blocked GSTA2 induction by t-BHQ, which was reversed by PMA-induced PKC activation. These results provide evidence that PKC may play a role in formation of peroxynitrite that activates Nrf2 for GSTA2 induction and that PKC may serve an activator for GSTA2 induction downstream of Pl3-kinase. PMID- 15357005 TI - The effect of vehicles and pressure sensitive adhesives on the percutaneous absorption of quercetin through the hairless mouse skin. AB - To investigate the feasibility of developing a new quercetin transdermal system, a preformulation study was carried out. Therefore, the effects of vehicles and pressure-sensitive adhesives (PSA) on the in vitro permeation of quercetin across dorsal hairless mouse skin were studied. Among vehicles used, propylene glycol monocaprylate (PGMC) and propylene glycol monolaurate were found to have relatively high permeation flux from solution formulation (i.e., the permeation fluxes were 17.25 +/- 1.96 and 9.60 +/- 3.87 microg/cm2/h, respectively). The release rate from PSA formulations followed a matrix-controlled diffusion model and was mainly affected by the amount of PSA and drug loaded. The overall permeation fluxes from PSA formulations were less than 0.30 microg/cm2/h, which were significantly lower compared to those obtained from solution formulations. The lower permeation fluxes may be due to the decrease of solubility and diffusivity of quercetin in the PSA layer, considering the fact that the highest flux of 0.26 microg/cm2/h was obtained with the addition of 0.2% butylated hydroxyanisole in PGMC-diethylene glycol monoethyl ether co-solvents (80-85 : 15 20, v/v). Taken together, these observations indicate that improvement in the solubility and diffusivity of quercetin is necessary to realize fully the clinically applicable transdermal delivery system for the drug. PMID- 15357006 TI - Expression of hepatic vascular stress genes following ischemia/reperfusion and subsequent endotoxemia. AB - Hepatic ischemia and reperfusion (I/R) predisposes the liver to secondary stresses such as endotoxemia, possibly via dysregulation of the hepatic microcirculation secondary to an imbalanced regulation of the vascular stress genes. In this study, the effect of hepatic I/R on the hepatic vasoregulatory gene expression in response to endotoxin was determined. Rats were subjected to 90 min of hepatic ischemia and 6 h of reperfusion. Lipopolysaccharide (LPS, 1 mg/kg) was injected intraperitoneally after reperfusion. Plasma and liver samples were obtained 6 h after reperfusion for serum aminotransferase assays and RT-PCR analysis of the mRNA for the genes of interest: endothelin-1 (ET-1), its receptors ET A and ET B, endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), cyclooxygenase-2 (COX-2), and tumor necrosis factor-alpha (TNF-alpha). The activities of serum aminotransferases were significantly increased in the I/R group. This increase was markedly potentiated by LPS treatment. The ET-1 mRNA was increased by LPS alone, and this increase was significantly greater in both the I/R alone and I/R + LPS groups compared to the sham. There were no significant differences in ET A receptor mRNA levels among any of the experimental groups. ET B mRNA was increased by both LPS alone and I/R alone, with no significant difference between the I/R alone and I/R + LPS groups. The eNOS and HO-1 transcripts were increased by I/R alone and further increased by I/R + LPS. The iNOS mRNA levels were increased by I/R alone, but increased significantly more by both LPS alone and I/R + LPS compared to I/R alone. The TNF-alpha mRNA levels showed no change with I/R alone, but were increased by both LPS alone and I/R + LPS. The COX-2 expression was increased significantly by I/R alone and significantly more by I/R + LPS. Taken collectively, significantly greater induction of the vasodilator genes over the constriction forces was observed with I/R + LPS. These results may partly explain the increased susceptibility of ischemic livers to injury as a result of endotoxemia. PMID- 15357007 TI - Cloning and characterization of directly amplified antiviral gene interferon alpha-2b (HulFNalpha-2b) from human leukocytes chromosomal DNA. AB - Interferons are cytokines that confer resistance to viral infection and inhibit cellular proliferation. The interferon alpha gene from human blood samples was amplified, cloned and expressed in E. coli (BL21). Leukocyte chromosomal DNA was used as a source of template DNA. Using specific primers, the gene for HulFNalpha 2b was amplified and inserted into the E. coli vector, pET21b, by ligation of the Hindlll and BamHI linkers of the vector and insert. The insert was further analyzed by PCR, DNA restriction mapping and sequencing, and expressed in a suitable E. coli strain. The production of this important cellular protein in the laboratory has significant applications in production of the recombinant pharmaceutical proteins. PMID- 15357008 TI - Pretreatment with 1,8-cineole potentiates thioacetamide-induced hepatotoxicity and immunosuppression. AB - The effect of 1,8-cineole on cytochrome P450 (CYP) expression was investigated in male Sprague Dawley rats and female BALB/c mice. When rats were treated orally with 200, 400 and 800 mg/kg of 1,8-cineole for 3 consecutive days, the liver microsomal activities of benzyloxyresorufin- and pentoxyresorufin-omicron dealkylases and erythromycin N-demethylase were dose-dependently induced. The Western immunoblotting analyses clearly indicated the induction of CYP 2B1/2 and CYP 3A1/2 proteins by 1,8-cineole. At the doses employed, 1,8-cineole did not cause toxicity, including hepatotoxicity. Subsequently, 1,8-cineole was applied to study the role of metabolic activation in thioacetamide-induced hepatotoxicity and/or immunotoxicity in animal models. To investigate a possible role of metabolic activation by CYP enzymes in thioacetamide-induced hepatotoxicity, rats were pre-treated with 800 mg/kg of 1 ,8-cineole for 3 days, followed by a single intraperitoneal treatment with 50 and 100 mg/kg of thioacetamide in saline. 24 h later, thioacetamide-induced hepatotoxicity was significantly potentiated by the pretreatment with 1,8-cineole. When female BALB/c mice were pretreated with 800 mg/kg of 1,8-cineole for 3 days, followed by a single intraperitoneal treatment with 100 mg/kg of thioacetamide, the antibody response to sheep red blood cells was significantly potentiated. In addition, the liver microsomal activities of CYP 2B enzymes were significantly induced by 1,8-cineole as in rats. Taken together, our results indicated that 1,8-cineole might be a useful CYP modulator in investigating the possible role of metabolic activation in chemical-induced hepatotoxicity and immunotoxicity. PMID- 15357009 TI - Wild ginseng prevents the onset of high-fat diet induced hyperglycemia and obesity in ICR mice. AB - Ginseng is a shade-loving perennial herb that is cultivated mainly in Korea, Japan, and China. The ginseng root has been used as a tonic remedy, and its antidiabetic activity has been demonstrated as early as 1920s. Although wild ginseng was anecdotally thought to be superior to cultivated ginseng as far as pharmacological properties were concerned, there have been no prior reports on the antidiabetic effect of wild ginseng. In this study, we investigated the preventative anti-diabetic and anti-obese effects of wild ginseng ethanol extract (WGEE). In the preventive experiment, WGEE co-administered with a high fat diet significantly inhibited body weight gain, fasting blood glucose, triglyceride, and free fatty acid levels in a dose dependent manner. WGEE-treated mice at doses of 250 and 500 mg/kg improved the insulin resistance index by 55% and 61% compared to the high fat diet (HFD) control, respectively. Diameters of white and brown adipocytes were also decreased by 62% and 46% in the WG500-treated group compared to those in HFD fed control mice. Taken together, WGEE has potential as a preventive agent for type 2 diabetes mellitus (and possibly obesity) and deserves clinical trial in the near future. PMID- 15357010 TI - Optimizing the novel formulation of liposome-polycation-DNA complexes (LPD) by central composite design. AB - LPD vectors are non-viral vehicles for gene delivery comprised of polycation condensed plasmid DNA and liposomes. Here, we described a novel anionic LPD formulation containing protamine-DNA complexes and pH sensitive liposomes composed of DOPE and cholesteryl hemisuccinate (Chems). Central composite design (CCD) was employed to optimize stable LPD formulation with small particle size. A three factor, five-level CCD design was used for the optimization procedure, with the weight ratio of protamine/DNA (X1), the weight ratio of Chems/ DNA (X2) and the molar ratio of Chems/DOPE in the anionic liposomes (X3) as the independent variables. LPD size (Y1) and LPD protection efficiency against nuclease (Y2) were response variables. Zeta potential determination was utilized to define the experimental design region. Based on experimental design, responses for the 15 formulations were obtained. Mathematical equations and response surface plots were used to relate the dependent and independent variables. The mathematical model predicted optimized X1-X3 levels that achieve the desired particle size and the protection efficiency against nuclease. According to these levels, an optimized LPD formulation was prepared, resulting in a particle size of 185.3 nm and protection efficiency of 80.22%. PMID- 15357011 TI - Pharmacokinetic-pharmacodynamic modeling for the relationship between glucose lowering effect and plasma concentration of metformin in volunteers. AB - Metformin is a biguanide antihyperglycemic agent often used for the treatment of non-insulin dependent diabetics (NIDDM). In this study, the pharmacokinetics and pharmacodynamics of metformin were investigated in Korean healthy volunteers during a fasting state for over 10 h. In order to evaluate the amount of glucose lowering effect of metformin, the plasma concentrations of glucose were measured for a period of 10 h followed by the administration of metformin (oral 500 mg) or placebo. In addition, the concentration of metformin in blood samples was determined by HPLC assay for the drug. All volunteers were consumed with 12 g of white sugar 10 minutes after drug intake to maintain initial plasma glucose concentration. The time courses of the plasma concentration of metformin and the glucose-lowering effect were analyzed by nonlinear regression analysis. The estimated Cmax, Tmax, CLt/F (apparent clearance), V/F(apparent volume of distribution), and half-life of metformin were 1.42 +/- 0.07 microg/mL, 2.59 +/- 0.18 h, 66.12 +/- 4.6 L/h, 26.63 L, and 1.54 h respectively. Since a significant counterclock-wise hysteresis was found for the metformin concentration in the plasma-effect relationship, indirect response model was used to evaluate pharmacodynamic parameters for metformin. The mean concentration at half-maximum inhibition IC50, kin, and kout, were 2.26 microg/mL, 83.26 h(-1), and 0.68 h(-1), respectively. Therefore, the pharmacokinetic-pharmacodynamic model may be useful in the description for the relationship between plasma concentration of metformin and its glucose-lowering effect. PMID- 15357012 TI - [Children and adolescents with hyperkinetic disorder. Frequency of the claims diagnosis in primary care based on the data of a regional Statutory Health Insurance Sample--Versichertenstichprobe AOK Hessen/KV Hessen (1998-2001)]. AB - OBJECTIVE: The study aims to estimate the frequency with which doctors of various groups submitted claims for the treatment of hyperkinetic disorder (HD; ICD-10) in children and adolescents in outpatient care in the period 1998-2001. METHOD: DATABASE: 18.75% random sample of a regional Statutory Health Insurance for the years 1998 until 2001 with n = 41,930 children and adolescents (1998: 0 to 15 years old). Children with HD were defined according to the wording of the diagnosis (1998, 1999) and the ICD 10-code: F90.0-F90.9 (2000, 2001). RESULTS: Between 1998 and 2001 the percentage of children with a claims diagnosis of HD rose from 1.6% to 2.4% (administrative prevalence). The male to female ratio is about four to one (2001, boys: 3.8%; girls: 1.0%). The highest rate is seen in the age group of 7 to 10 years (2001, boys: 5.8%; girls: 1.4%). The diagnosis was documented by pediatricians in 54% of the children with HD, by GPs in 29% and by child and adolescent psychiatrists in 15%. Since 1998 child and adolescent psychiatrists became more important in the treatment of children with HD. CONCLUSION: The data show an increase of the administrative prevalence of hyperkinetic disorder. The estimates are within the lower range of the prevalences gained by representative population-based field studies. PMID- 15357013 TI - [First effects of training phonological awareness in two age cohorts of kindergarten children]. AB - OBJECTIVE: This study investigated to what extent young kindergarten children already benefit from training phonological awareness. METHODS: The training program used here was the German "Wurzburg training program of phonological awareness". The control group participated in the regular kindergarten program. A total of 80 children, one and two years before entering school, participated in the study. Tests of phonological awareness were assessed before and after the training as well as after six months. RESULTS: Young kindergarten children (two years before entering school) compared to preschoolers (one year before entering school) had significant lower phonological awareness skills. However both cohorts showed noticeable significant training effects, but preschoolers benefited significantly more from training phonological awareness. Long-term effects were obtained for both cohorts. CONCLUSIONS: Already young kindergarten children do benefit effectively from preschool phonological awareness training. Effects of this training on reading and spelling in school will be investigated further. PMID- 15357014 TI - [Maternal smoking during pregnancy: risk factor for ADHD in the offspring?]. AB - OBJECTIVE: To examine the specifity of the association between maternal smoking during pregnancy and child behaviour problems. METHOD: In a prospective longitudinal study of a birth cohort of 362 children at risk parental tobacco use, cognitive and social-emotional child outcome and characteristics of family environment were assessed. RESULTS: About a quarter of mothers reported regular tobacco use during pregnancy. Children in this group exhibited enhanced rates of ADHD. This association held even after adjustment for a number of covariates. CONCLUSIONS: The findings suggest that maternal smoking during pregnancy may represent an independent risk factor for ADHD in the offspring. PMID- 15357015 TI - [Neurofeedback for the treatment of attention-deficit/hyperactivity disorder (ADHD) in childhood and adolescence]. AB - INTRODUCTION: A variety of non-pharmacological treatments for ADHD have failed to prove their effectiveness. The basis of evidence for neurofeedback (or EEG biofeedback) has improved recently. Neurofeedback is aiming at an improvement of ADHD core-symptoms via the voluntary modification of abnormal neurophysiologic parameters, e.g. EEG-frequency spectrum and event-related potentials. METHODS: Our review presents an overview of the current research on neurofeedback for the treatment of ADHD. RESULTS: In three controlled studies short-term effects of feedback matched those of stimulant treatment. Neurofeedback lead to significant improvement of attention, impulsivity and hyperactivity, without adversive side effects. Additionally, there was a persistent amelioration of EEG parameters, while stimulants did not lead to a comparable normalization. Results on the stability of feedback effects are encouraging but are based on small numbers of patients. CONCLUSION: Neurofeedback is a promising approach for the treatment of children with ADHD. However, there is a demand for further controlled studies using standardized diagnostic criteria, sufficient sample sizes and appropriate measures and follow-up. PMID- 15357016 TI - [Migrainous complex hallucinations in 10-year old patient--a case report and review]. AB - OBJECTIVES: Considering a prevalence of three to ten percent migraine in children is not a rare disease. Visual auras as a characteristic of classical migraine are often described in literature. However, most of these are primary hallucinations, whilst descriptions of complex hallucinations are barely to find. As far as we know this work describes the youngest age, at which complex hallucinations occurred in migraine attacks. Reviewing the existing literature possible pathophysiologic mechanisms and differential diagnosis are discussed. METHODS: We introduce a case-report of a 10-year old-patient who reports a complex visual hallucination during a migraine attack. Symptoms, history and diagnostic measures are described. In differential diagnosis to a migrainous aura the "Alice-in Wonderland-syndrome", "Lilliputian hallucinations", the basilar migraine and epilepsy are discussed. RESULTS: For complex hallucinations an analogical pathomechanism to primary hallucinations in migraine can be assumed. The great variety of the migrainous aura is pointed out. PMID- 15357018 TI - Cloning and characterization of a cDNA encoding diacylglycerol acyltransferase from castor bean. AB - The oil from castor seed (Ricinus communis) contains 90% ricinoleate, a hydroxy FA that is used in producing numerous industrial products. Castor diacylglycerol acyltransferase (RcDGAT) is a critical enzyme, as it catalyzes the terminal step in castor oil biosynthesis in which the products contain two or three ricinoleoyl moieties. We have isolated a cDNA encoding RcDGAT from developing castor seeds. Analysis of the sequence reveals that this cDNA encodes a protein of 521 amino acids with a molecular mass of 59.9 kDa. Although there are regions of high similarity to other plant DGAT coding sequences, there are sequences that distinguish it as well. Southern blot analysis suggests that the castor genome contains a single copy of RcDGAT. Analysis by reverse transcription-PCR reveals that the accumulation of the mRNA reaches its highest level at 19 d after pollination and declines thereafter. Expression of the full-length cDNA for RcDGAT in the yeast Saccharomyces cerevisiae, strain INVSc1 results in sevenfold higher DGAT activity compared with controls. When different molecular species of DAG were provided as substrates to the microsomal mixture, the RcDGAT showed a greater preference to catalyze the transfer of oleate from [14C]oleoyl-CoA to diricinolein than to diolein and dipalmitolein. With the addition of 0.25 mM substrates, diricinolein gave 318 pmol/mg/min diricinoleoyloleoylglycerol (RRO), while diolein and dipalmitolein gave only about 195 pmol/mg/min of triolein (OOO) and 120 pmol/mg/min dipalmitoyleoylglycerol (PoPoO), respectively. This work will facilitate investigation of the role of RcDGAT in castor oil biosynthesis. PMID- 15357019 TI - Biosynthesis and isomerization of 11-hydroperoxylinoleates by manganese- and iron dependent lipoxygenases. AB - Manganese lipoxygenase (Mn-LO) oxygenates linoleic acid (LA) to a mixture of the hydroperoxides--11 (S)-hydroperoxy-9Z,12Z-octadecadienoic acid [11(S)-HPODE] and 13(R)-hydroperoxy-9Z,11 E-octadecadienoic acid [13(R)-HPODE]-- and also catalyzes the conversion of 11 (S)-HPODE to 13(R)-HPODE via oxygen-centered (LOO-) and carbon-centered (L.) radicals [Hamberg, M., Su, C., and Oliw, E. (1998) Manganese Lipoxygenase. Discovery of a Bis-allylic Hydroperoxide as Product and Intermediate in a Lipoxygenase Reaction, J. Biol. Chem. 273, 13080-13088]. The aims of the present work were to investigate whether 11-HPODE can also be produced by iron-dependent lipoxygenases and to determine the enzymatic transformations of stereoisomers of 11-HPODE by lipoxygenases. Rice leaf pathogen inducible lipoxygenase, but not soybean lipoxygenase-1 (sLO-1), generated a low level of 11-HPODE (0.4%) besides its main hydroperoxide, 13(S)-HPODE, on incubation with LA. Steric analysis revealed that 11-HPODE was enriched with respect to the R enantiomer [74% 11(R)]. In agreement with previous results, 11 (S)-HPODE incubated with Mn-LO provided 13(R)-HPODE, and the same conversion also took place with the methyl ester of 11(S)-HPODE. 11(R,S)-HPODE was metabolized biphasically in the presence of Mn-LO, i.e., by a rapid phase during which the 11(S)-enantiomer was converted into 13(R)-HPODE and a slow phase during which the 11(R)-enantiomer was converted into 9(R)-HPODE. sLO-1 catalyzed a slow conversion of 11 (S)-HPODE into a mixture of 13(R)-HPODE (75%), 9(S)-HPODE (10%), and 13(S) HPODE (10%), whereas 11(R,S)-HPODE produced a mixture of nearly racemic 13-HPODE (approximately 70%) and 9-HPODE (approximately 30%). The results showed that 11HPODE can also be produced by an iron-dependent LO and suggested that the previously established mechanism of isomerization of 11(S)-HPODE involving suprafacial migration of O2 is valid also for the isomerizations of 11(R)-HPODE by Mn-LO and of 11(S)-HPODE by sLO-1. PMID- 15357017 TI - Isoprenoids: remarkable diversity of form and function. AB - The isoprenoid biosynthetic pathway is the source of a wide array of products. The pathway has been highly conserved throughout evolution, and isoprenoids are some of the most ancient biomolecules ever identified, playing key roles in many life forms. In this review we focus on C-10 mono-, C-15 sesqui-, and C-20 diterpenes. Evidence for interconversion between the pathway intermediates farnesyl pyrophosphate and geranylgeranyl pyrophosphate and their respective metabolites is examined. The diverse functions of these molecules are discussed in detail, including their ability to regulate expression of the beta-HMG-CoA reductase and Ras-related proteins. Additional topics include the mechanisms underlying the apoptotic effects of select isoprenoids, antiulcer activities, and the disposition and degradation of isoprenoids in the environment. Finally, the significance of pharmacological manipulation of the isoprenoid pathway and clinical correlations are discussed. PMID- 15357020 TI - Dose effect of alpha-linolenic acid on PUFA conversion, bioavailability, and storage in the hamster. AB - If an increased consumption of alpha-linolenic acid (ALA) is to be promoted in parallel with that of n-3 long-chain-rich food, it is necessary to consider to what extent dietary ALA can be absorbed, transported, stored, and converted into long-chain derivatives. We investigated these processes in male hamsters, over a broad range of supply as linseed oil (0.37, 3.5, 6.9, and 14.6% energy). Linoleic acid (LA) was kept constant (8.5% energy), and the LA/ALA ratio was varied from 22.5 to 0.6. The apparent absorption of individual FA was very high (>96%), and that of ALA remained almost maximum even at the largest supply (99.5%). The capacity for ALA transport and storage had no limitation over the chosen range of dietary intake. Indeed, ALA intake was significantly correlated with ALA level not only in cholesteryl esters (from 0.3 to 9.7% of total FA) but also in plasma phospholipids and red blood cells (RBC), which makes blood components extremely reliable as biomarkers of ALA consumption. Similarly, ALA storage in adipose tissue increased from 0.85 to 14% of total FA and was highly correlated with ALA intake. As for bioconversion, dietary ALA failed to increase 22:6n-3, decreased 20:4n-6, and efficiently increased 20:5n-3 (EPA) in RBC and cardiomyocytes. EPA accumulation did not tend to plateau, in accordance with identical activities of delta5- and delta6-desaturases in all groups. Dietary supply of ALA was therefore a very efficient means of improving the 20:4n-6 to 20:5n-3 balance. PMID- 15357021 TI - Type 1 diabetes compromises plasma arachidonic and docosahexaenoic acids in newborn babies. AB - The activity of delta6- and delta5-desaturase, enzymes required for the synthesis of AA and DHA, are impaired in human and experimental diabetes. We have investigated whether neonates of type 1 diabetic women have compromised plasma AA and DHA at birth. Cord blood was obtained from healthy babies born to mothers with (n = 31) and without (n = 59) type 1 diabetes. FA composition of plasma choline phosphoglycerides (CPG), TG, and cholesterol esters (CE) was assayed. The neonates of the diabetics had lower levels of AA (20:4n-6, P< 0.0001), adrenic acid (22:4n-6, P < 0.01), sigman-6 metabolites (P < 0.0001), docosapentaenoic acid (22:5n-3, P < 0.0001), DHA (22:6n-3, P < 0.0001), sigman-3 (P < 0.0001), and sigman-3 metabolites (P< 0.0001) in CPG compared with the corresponding babies of the nondiabetic mothers. Similarly, they had lower levels of AA (P< 0.05), sigman 6 metabolites (P < 0.05), DHA (P< 0.0001), and sigman-3 metabolites (P< 0.01) in plasma CE. There was also a nonsignificant reduction of AA and DHA in TG in the babies of the diabetic group. The current investigation indicates that healthy neonates born to mothers with type 1 diabetes have highly compromised levels of AA and DHA. These nutrients are of critical importance for neurovisual and vascular system development. In poorly controlled maternal diabetes, it is conceivable that the relative "insufficiency" of AA and DHA may exacerbate speech and reading impairments, behavioral disorders, suboptimal performance on developmental tests, and lower IQ, which have been reported in some children born to mothers with type 1 diabetes mellitus. Further studies are needed to understand the underlying mechanism for this biochemical abnormality and its implications for fetal and infant development. PMID- 15357022 TI - An isocaloric PUFA diet enhances lipid uptake and weight gain in aging rats. AB - Aging is associated with a change in the morphology and absorptive capacity of the small intestine. In young rats, feeding a semisynthetic diet containing saturated FA (SFA) increases nutrient uptake, as compared with an isocaloric diet containing polyunsaturated FA (PUFA). We tested the hypotheses that (i) aging is associated with a decline in lipid absorption in the Fischer 344 rat; (ii) this decline can be corrected by manipulating the fat composition of the diet; and (iii) the age- and diet-associated variations in lipid uptake are associated with changes in the ileal lipid-binding protein (ILBP) or the intestinal or liver FA binding proteins (I- or L-FABP, respectively) in the cytosol of the enterocyte. In rats fed SFA or PUFA, aging was associated with a decline in the in vitro uptake of stearic acid (18:0) when expressed on the basis of intestinal or mucosal weight. In contrast, age had no effect on lipid uptake when expressed on the basis of serosal surface area, whereas lipid uptake increased with age when expressed on the basis of mucosal surface area. The age-associated variations in lipid uptake were not associated with changes in protein abundance and/or expression of ILBP, I-FABP, or L-FABP. In 24-mon-old rats, when uptake of lipids was expressed on the basis of mucosal surface area, feeding PUFA enhanced lipid uptake and body weight gain as compared with rats fed SFA. Future studies must determine whether the enhanced lipid uptake and body weight gain observed in older animals fed PUFA have any therapeutic benefit. PMID- 15357023 TI - CLA isomers in milk fat from cows fed diets with high levels of unsaturated fatty acids. AB - The concentrations of CLA isomers were determined by Ag+ -HPLC in the milk fat of cows fed a control diet consisting of hay ad libitum and 15 kg of fodder beets or this diet supplemented with oilseeds containing either high levels of oleic acid (rapeseed), linoleic acid (sunflower seed), or alphalinolenic acid (linseed). Highly significant (P < or = 0.001) correlations were found between the daily intakes of oleic acid and the concentration of the CLA isomer trans-7,cis-9 in milk fat; of linoleic acid and the CLA isomers trans-10,trans-12, trans-9,trans 11, trans-8,trans-10, trans-7,trans-9, trans-10,cis-12, cis-9,trans-11, trans 8,cis-10, and trans-7,cis-9; and of alpha-linolenic acid and the CLA isomers trans-12,trans-14, trans-11 ,trans-13, cis,trans/trans,cis-12,14, trans-11 ,cis 13, and cis-11 ,trans-13. CLA concentrations were also determined in the milk fat of cows grazing in the lowlands (600-650 m), the mountains (900-1210 m), and the highlands (1275-2120 m). The concentrations of many isomers were highest in milk fat from the highlands, but only three CLA isomers (cis-9,trans-11, trans-11 ,cis 13, and trans-8,cis-10) showed a nearly linear increase with elevation. Therefore, these three CLA isomers, and particularly the CLA isomer trans- 11,cis 13, the second-most important CLA in milk fat from cows grazing at the three altitudes, could be useful indicators of milk products of Alpine origin. PMID- 15357024 TI - Effect of CLA on milk fat synthesis in dairy cows: comparison of inhibition by methyl esters and free fatty acids, and relationships among studies. AB - CLA is a potent inhibitor of milk fat synthesis, as shown by investigations using mixtures of CLA isomers in FFA form. However, methyl esters of CLA can be initially formed in commercial synthesis, and their use in a supplement has certain manufacturing and cost advantages. Our objective was to compare abomasal infusion of methyl esters of CLA (ME-CLA) and FFA of CLA (FFA-CLA) on milk fat synthesis. Data were also combined with previous investigations to examine broader relationships between trans-10,cis-12 CLA and the reduction in milk fat. Three mid-lactation, rumen-fistulated Holstein cows were used in a 3 x 3 Latin square design. Treatments were (i) control, (ii) ME-CLA, and (iii) FFA-CLA. The ME-CLA and FFA-CLA treatments (4.2 g/d trans-10,cis-12 CLA) resulted in a comparable reduction in milk fat yield (38 and 39%, respectively) and pattern of reduction in individual FA. In contrast, milk yield, milk protein, and feed intake were unaltered by CLA treatment. Combining data across studies revealed strong correlations relating the reduction in milk fat yield to abomasal dose of trans-10,cis-12 CLA (R2 = 0.86), milk fat content of trans-10,cis-12 CLA (R2 = 0.93), and milk fat secretion of trans-10,cis-12 CLA (R2 = 0.82). Across studies, transfer efficiency of abomasally infused trans-10,cis-12 CLA into milk fat was relatively constant (22%; R2 = 0.94). Overall, ME-CLA and FFA-CLA were equally potent in reducing milk fat, and either form could be used to formulate a dietary supplement that would induce milk fat depression. PMID- 15357025 TI - Positional distribution of decanoic acid: effect on chylomicron and VLDL TAG structures and postprandial lipemia. AB - Although medium-chain FA (MCFA) are mainly absorbed via the portal venous system, they are also incorporated into chylomicron TAG; therefore, the positional distribution of MCFA in TAG is likely to affect their metabolic fate. We studied chylomicron and VLDL TAG structures, as well as the magnitude of postprandial lipemia, after two oral fat loads containing decanoic acid (10:0) predominantly at the sn-1(3),2 (MML) or at the sn-1,3 positions (MLM) of TAG in a randomized, double-blind, crossover clinical trial with 10 healthy, normal-weight volunteers. An MS-MS method was used to analyze TAG regioisomers. The position of decanoic acid in chylomicron TAG reflected its position in the TAG ingested, and TAG with none, one, two, or three decanoic acid residues were detected after ingestion of both fats. More (P < 0.05) 30:0 and 38:1 TAG (acyl carbons:double bonds) and fewer 46:5, 54:5, and 54:4 TAG were found in chylomicrons after ingestion of MML than after MLM. The VLDL TAG composition did not differ between the fat loads but did change (P < 0.05) 2 to 6 h after ingestion of both fats. No statistical differences were seen between the fat loads in areas under the plasma, chylomicron, or VLDL TAG response curves or in FFA concentrations. Thus, the positional distribution of MCFA in TAG affects their metabolic fate, but the magnitude of postprandial lipemia does not seem to be dependent on the positional distribution of MCFA in the ingested fat. PMID- 15357026 TI - Selective increase in pinolenic acid (all-cis-5,9,12-18:3) in Korean pine nut oil by crystallization and its effect on LDL-receptor activity. AB - The aims of this study were to obtain concentrated pinolenic acid (5,9,12-18:3) from dietary Korean pine (Pinus koraiensis) nut oil by urea complexation and to investigate its cholesterol-lowering effect on the LDL-receptor activity of human hepatoma HepG2 cells. Pine nut oil was hydrolyzed to provide a low-pinolenic acid containing FA extract (LPAFAE), followed by crystallization with different ratios of urea in ethanol (EtOH) or methanol (MeOH) as a solvent to produce a high pinolenic acid-containing FA extract (HPAFAE). The profiles of HPAFAE obtained by urea complexation showed different FA compositions compared with LPAFAE. The long chain saturated FA palmitic acid (16:0) and stearic acid (18:0) were decreased with urea/FA ratios (UFR) of 1:1 (UFR1), 2:1 (UFR2), and 3:1 (UFR3). Linoleic acid (9,12-18:2) was increased 1.3 times with UFR2 in EtOH, and linolenic acid (9,12,15-18:3) was increased 1.5 times with UFR3 in MeOH after crystallization. The crystallization with UFR3 in EtOH provided the highest concentration of pinolenic acid, which was elevated by 3.2-fold from 14.1 to 45.1%, whereas that of linoleic acid (9,12-18:2) was not changed, and that of oleic acid (9-18:1) was decreased 7.2-fold. Treatment of HepG2 cells with HPAFE resulted in significantly higher internalization of 3,3'-dioctadecylindocarbocyanine-LDL (47.0 +/- 0.15) as compared with treatment with LPAFAE (25.6 +/- 0.36) (P< 0.05). Thus, we demonstrate a method for the concentration of pinolenic acid and suggest that this concentrate may have LDL-lowering properties by enhancing hepatic LDL uptake. PMID- 15357027 TI - Fenitrothion-induced structural and functional perturbations in the yolk lipoproteins of the shrimp Macrobrachium borellii. AB - Two lipovitellin (LV) forms containing the same apoproteins but differing in their lipid composition were isolated from Macrobrachium borelii eggs at early (LVe) and late (LVI) embryogenic stages and characterized. These two forms of LV, as well as liposomes prepared with lipids extracted from them, were used as simpler models to study the effect of the pesticide fenitrothion (FS) on their structures and functions. Rotational diffusion and fluorescence lifetime of two fluorescent probes [1,6-diphenyl-1,3,5-hexatriene (DPH) and 3-(p-(6-phenyl)-1,3,5 hexatrienal)phenylpropionic acid (DPH-PA)] were used to obtain information on structural changes induced by FS in the inner and outer regions of the LV, respectively. Comparison of the rotational behavior of these probes in native LV and liposomes (LP) from extracted LV lipids suggests that apoprotein-lipid interactions result in an ordered neutral lipid core. FS increased the lipid phase polarity of both LV and LP forms. The rotation of these probes in LP was not affected, suggesting a dependence of FS action on lipid-protein interactions. DPH-PA steady-state anisotropy showed that, unlike the LVe form, the LVI form was sensitive to extremely low FS concentrations. The ability of both LV to transfer palmitic acid to albumin was increased, but in a dissimilar manner, by the presence of FS. Such differences in the sensitivity of the LV at different steps of embryogenesis to FS influence the toxic action of this insecticide. PMID- 15357028 TI - Synthesis of deuterated fatty acids to investigate the biosynthetic pathway of disparlure, the sex pheromone of the gypsy moth, Lymantria dispar. AB - The preparation and characterization of a series of deuterium-labeled intermediates used in the study of the biosynthetic pathway for disparlure, the sex pheromone of Lymantria dispar, is reported. The synthetic route starts with propargyl alcohol, and the deuterium atoms are introduced by deuteration of an alkyne precursor in the presence of Wilkinson's catalyst. The olefinic bond was created by the Wittig reaction of a suitable aldehyde with a common tetradeuterated phosphonium ylide intermediate. The presence of the expected label and its correct location were confirmed by both MS and 13C NMR. These compounds were successfully used to elucidate the disparlure biosynthetic pathway. PMID- 15357029 TI - Robert Holley--1968 Nobel Laureate. PMID- 15357030 TI - Peripheral arterial disease in the catheterization laboratory: an underdetected and undertreated risk factor. PMID- 15357031 TI - Evaluation and epidemiological research in tuberculosis control: linking medical care and public health. PMID- 15357032 TI - Effect of peripheral arterial disease in patients undergoing percutaneous coronary intervention with intracoronary stents. AB - OBJECTIVE: To compare the short-term and long-term outcomes of patients with coronary artery disease and peripheral arterial disease (PAD) who underwent intracoronary (IC) stent Implantation during percutaneous coronary intervention (PCI) with the outcomes of patients with isolated coronary artery disease but without PAD who underwent IC stent implantation. PATIENTS AND METHODS: We analyzed the outcomes of 7696 patients who underwent IC stent implantation during PCI at the Mayo Clinic in Rochester, Minn, between January 1996 and December 2002. Outcomes of 6299 patients (82%) with Isolated coronary artery disease and without PAD who underwent IC stent implantation (group 1) were compared with outcomes of 1397 patients (18%) with coronary artery disease and PAD (group 2) who underwent PCI with IC stent implantation. RESULTS: Patients in group 2 were older (71.1+/-10.2 years vs 65.0+/-12.0 years; P<.001) and had a higher prevalence of hypertension (79% vs 61%; P<.001), diabetes mellitus (33% vs 20%; P<.001), hyperlipidemia (76% vs 70%; P<.001), and history of smoking (70% vs 63%; P<.001) compared with group 1. Prevalence of multivessel disease was higher in group 2 (79% vs 68%; P<.001). Procedural success was significantly lower In group 2 (95% vs 97%; P<.001). In-hospital complications were higher in group 2: death (3% vs 1%; P<.001), any myocardial Infarction (MI) (8% vs 5%; P<.001), death/MI/coronary artery bypass grafting (CABG)/target vessel revascularization (11% vs 7%; P<.001), and blood loss requiring transfusion (11% vs 5.8%; P<.001). After adjustment for other risk factors, the odds ratio for in-hospital death was 1.84 (95% confidence interval [CI], 1.16-2.90; P=.009), and for death/MI/CABG/target vessel revascularization, the odds ratio was 1.25 (95% CI, 1.00-1.55; P=.048) in patients with PAD treated with IC stents. Median follow-up was 3.1 years. Six-month, 1-year, and 2-year Kaplan-Meier estimates of survival free of death/MI/CABG/target vessel revascularization were 84%, 77%, and 69%, respectively, for group 2 and were significantly worse compared with group 1 (89%, 85%, and 80%, respectively; P<.001). This effect remained after adjustment for other risk factors (hazard ratio, 1.36; 95% CI, 1.22-1.51). CONCLUSIONS: Compared with patients who had isolated coronary artery disease but no PAD, patients with coronary artery disease and PAD had lower procedural success and higher in-hospital major cardiovascular complications, including higher blood loss requiring transfusion, after PCI with stent Implantation. On follow-up, the short-term and long-term outcomes of patients with PAD were worse, with higher mortality, MI, and need for repeated target vessel revascularization. PMID- 15357033 TI - Early life risk factors for attention-deficit/hyperactivity disorder: a population-based cohort study. AB - OBJECTIVES: To Identify risk factors for attention-deficit/hyperactivity disorder (ADHD) and to determine whether these factors differ between boys and girls. SUBJECTS AND METHODS: We conducted a case-control study of all children born between January 1, 1976, and December 31, 1982, in Olmsted County, Minnesota (N=5701), and examined birth certificates for risk factors for ADHD. Using school and medical records, we identified 305 children with ADHD. All children not identified with ADHD served as controls (N=5326). Risk factors that were assessed included demographic characteristics of both children and parents, as well as pregnancy, labor, and delivery characteristics, including the effect of a multiple birth and presence of pregnancy and delivery complications. RESULTS: Pregnancy and labor characteristics, low birth weight, and presence of a twin birth were not associated with ADHD. Male sex (odds ratio [OR], 3.05; 95% confidence interval [CI], 2.34-3.98) was associated with an increased risk for ADHD, whereas high maternal and paternal education levels (high maternal education: OR, 0.57; 95% CI, 0.42-0.78; high paternal education: OR, 0.58; 95% CI, 0.49-0.78) were associated with a decreased risk for ADHD. In addition, low maternal and paternal education levels increased the risk for ADHD in boys more than in girls. CONCLUSION: Male sex and low parental education levels are significant risk factors for ADHD. In addition, boys born to parents with low maternal and paternal education levels appear to be at an increased risk for ADHD compared with girls born to parents with low education levels. PMID- 15357034 TI - Robot-assisted laparoscopic cholecystectomy: initial Mayo Clinic Scottsdale experience. AB - OBJECTIVE: To review a single institution's experience with robot-assisted laparoscopic cholecystectomy. PATIENTS AND METHODS: We reviewed retrospectively all cholecystectomies performed with the assistance of a surgical robot at the Mayo Clinic in Scottsdale, Ariz, from October 2002 to July 2003. Cholanglograms were obtained intraoperatively for patients with elevated results on liver function tests. The gallbladder was removed by the surgeon with the use of the robot. A surgical assistant at the operating table exchanged instruments in the robot arms and applied clips when needed. Total operating time, assembly time of the robot, complications, and postoperative course were evaluated. RESULTS: Nineteen patients underwent robot-assisted laparoscopic cholecystectomy; 16 had symptomatic cholelithiasis, 2 had gallbladder polyps, and 1 had acute cholecystitis. Of the 19 surgeries, 16 were completed successfully with robotic assistance. In 3 consecutive patients, a mechanical problem occurred with the robot; however, all procedures were completed laparoscopically. The mean set-up time, including patient positioning and preparation and robotic installation, was 28.1 minutes. The mean +/- SD operating time was 82.3+/-17.9 minutes without a cholanglogram and 102+/-20.9 minutes with a cholanglogram. There were no complications and no conversions to an open procedure. CONCLUSION: Robotic surgery offers many potential advantages, including surgeon comfort, elimination of surgeon tremor, improved imaging, and increased degrees of freedom of the operative Instruments, compared with conventional laparoscopic surgery. However, patient outcomes and operative costs need to be evaluated further. PMID- 15357035 TI - Linezolid therapy for orthopedic infections. AB - OBJECTIVE: To assess the clinical and bacteriological efficacy and the safety and tolerability profile of linezolid when used as the primary component of a combined surgical and antimicrobial treatment strategy for orthopedic infections. PATIENTS AND METHODS: We retrospectively reviewed medical records to obtain clinical, bacteriological, and safety outcomes of 20 consecutive patients who took linezolid for the treatment of orthopedic infections between April 1, 2000, and November 30, 2002. RESULTS: Twenty patients received linezolid therapy for microbiologically confirmed orthopedic infections due to gram-positive cocci during the 32-month study period. Fifteen patients (75%) had infections involving orthopedic hardware; in 10 of these patients, the hardware was removed. All 20 patients underwent surgical debridement. At a mean follow-up of 276 days, 11 patients (55%) achieved clinical cure, 7 (35%) had clinical improvement but received long-term antimicrobial suppressive therapy, 1 (5%) had clinical relapse after discontinuing linezolid treatment, and 1 (5%) died of a cause unrelated to linezolid treatment. Bacterial persistence was documented in 3 patients (15%), all of whom were Infected with methicillin-resistant Staphylococcus aureus. Eight patients (40%) developed reversible myelosuppression, 1 (5%) had irreversible peripheral neuropathy, and 2 (10%) discontinued linezolid treatment because of pancytopenia or urticaria. CONCLUSION: Oral linezolid may be an effective alternative therapy for orthopedic infections due to linezolid-susceptible gram positive bacteria in patients who are unable to take other antimicrobial drugs because of drug allergy or Intolerance or antimicrobial resistance. PMID- 15357036 TI - Motivating factors for academic physicians within departments of medicine. AB - OBJECTIVE: To understand what motivates academic physicians at a time when physician dissatisfaction is prevalent. SUBJECTS AND METHODS: Of a cohort of 480 physician faculty members (identified from the Association of American Medical Colleges faculty roster) hired at the assistant professor level, 183 were monitored prospectively for a characterization of their success in achieving promotion. In mid-2001, follow-up data were collected about the factors that physicians described as motivating in their work. We conducted this study to understand the differences in motivators between clinician-educators and cliniclan-investigators and between male and female physicians, as well as to validate a previously used instrument developed to assess motivation and occupational values. RESULTS: Of 183 physicians monitored, 144 (79%) responded to an interim follow-up questionnaire. Factor analysis revealed that physicians' occupational motivators could be grouped statistically into 3 factors: self expression, helping others, and extrinsic rewards. Compared with clinician educators, clinician-investigators were more motivated in their current work by having the ability to express themselves (composite factor score, 4.30 vs 3.84; P<.001). Clinician-investigators also rated 4 of the 6 items within the factor of self-expression as being significantly stronger motivators for them than did the clinician-educators. Compared with male physicians, female physicians Indicated they were more motivated by helping others (composite factor score, 4.18 vs 3.89; P=.03). CONCLUSIONS: Factors that motivate physicians appear to be different for clinician-investigators and clinician-educators as well as for male and female physicians. Understanding the inspiration for physicians may help medical leadership to better motivate and relate to their physician workforce. PMID- 15357037 TI - Pulmonary involvement in Henoch-Schonlein purpura. AB - OBJECTIVE: To describe pulmonary involvement in Henoch-Schonlein purpura (HSP). PATIENTS AND METHODS: We retrospectively reviewed the medical records of patients with HSP who were seen at the Mayo Clinic in Rochester, Minn, during a 6-year period (January 1, 1997, to December 31, 2002). Patients with HSP and pulmonary involvement were identified through a review of clinical records, radiological studies, pulmonary function data, and lung biopsy findings. RESULTS: We identified 124 patients with HSP during the study period; 72 (58.1%) were males. The median age was 15 years (range, 8 months to 81 years). Among the 124 patients with HSP, 3 (2.4%), all adults, had pulmonary involvement. Of these 3 patients, aged 20, 67, and 76 years, 2 were women. Pulmonary manifestations consisted of diffuse alveolar hemorrhage (DAH) (2 patients) and usual interstitial pneumonia that improved with corticosteroid therapy (1 patient). CONCLUSIONS: Pulmonary involvement in HSP is rare. It occurs more often in adults and commonly manifests as DAH and occasionally as usual interstitial pneumonia or interstitial fibrosis. Our cases and previously reported cases suggest that DAH is the most common manifestation of pulmonary involvement in HSP. PMID- 15357038 TI - Primary care perspective on bariatric surgery. AB - The role of primary care physicians in identifying potential candidates for bariatric surgery and providing them with long-term postoperative care is gaining rapidly in importance. With the increased use of surgical procedures as treatment options for obesity, a knowledge of bariatric surgery is essential for providing optimal care. During the past decade, the number of bariatric procedures has increased, and refinements of these procedures have made them safer and more effective. Primary care physicians should know how to Identify appropriate surgical candidates and be familiar with available procedures, aware of potential complications and benefits, and able to provide lifelong monitoring and follow-up care. Thus, the primary care physician must be informed about surgical criteria, types of procedures, outcomes, complications, and the long-term monitoring needs of these patients. PMID- 15357039 TI - Sternal hump. PMID- 15357040 TI - Minimally invasive radical prostatectomy. AB - The development of advanced laparoscopic techniques and robot-assisted technology has resulted in several new surgical approaches for treating organ-confined prostate cancer. Outcomes with these new or minimally Invasive techniques should be assessed carefully to ensure that they are similar to or surpass patients' oncologic and functional outcomes after open radical prostatectomy. This article reviews the current published experience with minimally Invasive approaches to increase awareness about viability. Several of the larger series of patients who have undergone laparoscopic (transperitoneal and extraperitoneal) or robot assisted laparoscopic radical prostatectomies are discussed and evaluated critically. Comparisons to published data on open radical prostatectomy are included for completeness. The different minimally invasive techniques are described and contrasted in regard to prostate-specific antigen progression-free survival, surgical margin status, blood loss, transfusion rates, postoperative pain, length of hospitalization, duration of urinary catheterization, potency, continence, and complications. The relative costs of each method are provided. The coexistence of multiple surgical approaches should and can challenge surgeons who perform open and minimally invasive procedures to strive for a new standard of care above and beyond what is accepted today to minimize patient morbidity while maximizing functional and oncologic outcomes. PMID- 15357041 TI - 80-year-old man with anemia and blistering skin lesions. PMID- 15357042 TI - Vestibular toxicity due to inhaled tobramycin in a patient with renal insufficiency. AB - Inhaled tobramycin is being used increasingly in patients with cystic fibrosis and other forms of bronchiectasis for treatment of bronchial colonization with Pseudomonas aeruginosa. The goal of inhaled antibiotics is to provide maximal concentrations at the site of infection without risking systemic toxicity. We report an unusual case of reversible vestibular toxicity due to inhaled tobramycin in a patient with renal failure who was undergoing hemodialysis. Although systemic absorption after inhaled tobramycin is reportedly negligible, no recommendations have been published regarding monitoring of serum concentrations in patients receiving this form of therapy. We suggest that clinicians consider monitoring serum concentrations of tobramycin in patients at risk of renal toxicity and/or ototoxicity, such as those with predisposing renal or otologic compromise. Further studies in at-risk patients are needed to determine the optimal frequency and timing of such monitoring. PMID- 15357043 TI - Rosacea. PMID- 15357044 TI - Stevens-Johnson syndrome after immunization with smallpox, anthrax, and tetanus vaccines. AB - A 19-year-old male military recruit developed erythema multiforme 20 days after receiving a triad of vaccinations: smallpox (vaccinia virus), anthrax, and tetanus. Over the course of a few days, the erythema multiforme evolved into Stevens-Johnson syndrome, associated with widespread bullae, stomatitis, conjunctivitis, and fever. After 7 days of conservative management, the patient's signs and symptoms improved. This case serves as a timely reminder of a severe and potentially life-threatening complication of smallpox vaccination. PMID- 15357045 TI - Carotid revascularization for prevention of stroke: carotid endarterectomy and carotid artery stenting. AB - Carotid endarterectomy (CEA) has been used for the past several decades in patients with carotid occlusive disease. Large randomized controlled trials have documented that CEA is a highly effective stroke preventive among patients with carotid stenosis and recent transient ischemic attack or cerebral infarction. In asymptomatic patients with carotid stenosis, clinical trial data suggest that the degree of stroke prevention from CEA is less than among symptomatic patients. However, otherwise healthy men and women with an asymptomatic carotid stenosis of 60% or greater have a lower risk of future cerebral infarction, including disabling cerebral infarction, if treated with CEA compared with those treated with medical management alone. More recently, carotid artery stenting has been performed Increasingly for patients with carotid occlusive disease. As technology has improved, procedural risks have declined and are approaching those reported for CEA. The benefits and durability of CEA compared with carotid artery stenting are still unclear and are being studied in ongoing randomized controlled trials. PMID- 15357046 TI - Medicare reform needed for home-based low-molecular-weight heparin therapy. PMID- 15357047 TI - Celiac disease serology and irritable bowel syndrome: does the relationship merit further evaluation? PMID- 15357048 TI - Unenabled embryo use. PMID- 15357049 TI - Unenabled embryo use. PMID- 15357050 TI - Unenabled embryo use. PMID- 15357051 TI - The effect of parasitism on host fecundity is dependent on temperature in a cockroach-acanthocephalan system. AB - The fitness of infected organisms can vary greatly depending on the temperature at which they find themselves. Understanding the role of temperature in the fitness of infected organisms can be crucial to population studies, epidemiological studies, and when screening for biological control agents. We measured the effect of parasitism on host survival and reproduction at 4 constant temperatures using the acanthocephalan parasite Moniliformis moniliformis and its intermediate host, the cockroach Supella longipalpa. Infection did not affect cockroach survival at any temperature. Infection had a negative impact on cockroach fecundity but only at higher temperatures (28 and 31 C) and only later in infection (>20 days postinfection). At lower temperatures, infected and uninfected cockroaches had similar fecundities throughout the duration of the experiment (120 days). This study, along with previous studies, suggests that researchers would do well to consider environmental variables when exploring the effects of parasitism. PMID- 15357052 TI - Prevalence, intensity, and differential development of Pseudodelphis oligocotti (Nematoda: Dracunculoidea) in sympatric fish hosts of the northeastern Pacific coast. AB - Counter to expectations of coevolved parasite-host relationships, parasites frequently infect hosts that never contribute to their reproduction, making the identification of a parasite's true host-specificity problematic. Pseudodelphis oligocotti (Nematoda: Dracunculoidea) infects several coastal Pacific fishes, but its course of development appears highly variable, suggesting that incidence does not reflect effective host range. To determine the host range of P. oligocotti and describe its relationship to various potential hosts, 24 fish species were examined from several British Columbia localities for prevalence, intensity, and extent and tissue location of parasite development. Pseudodelphis oligocotti infects 9 species of fishes from 5 orders, of which penpoint gunnel, Apodichthys flavidus, showed the highest prevalence and intensity, up to 80% and 19 (+/- 17.1 SD) worms per host, respectively. Although subadult and adult P. oligocotti occurred in all 9 fishes, larvigerous P. oligocotti only occurred in A. flavidus and rarely in the northern clingfish, Gobiesox maeandricus. Infective first-stage larvae were recovered from gill tissue of A. flavidus. Thus, at most only 2 of the 9 host species infected by P. oligocotti actually contribute to its transmission. The occurrence of P. oligocotti in diverse hosts may be accounted for by the parasite's indiscriminant mode of transmission via ingestion of free living intermediate copepod hosts, where highly exposed or more suitable fishes (or both) are closely related by diet and microhabitat. This study demonstrates how parasite transmission and host ecology can greatly affect observed host range and ultimately its potential for expansion. PMID- 15357053 TI - Rio Meta strain of Plasmodium vivax in New World monkeys and anopheline mosquitoes. AB - An archived strain of Plasmodium vivax, isolated from Rio Meta, northern Colombia, in 1972 was adapted to grow in splenectomized Aotus lemurinus griseimembra and A. nancymai monkeys. Anopheles freeborni, An. maculatus, An. dirus, An. culicifacies, and An. albimanus were shown to be susceptible to infection by feeding on infected monkeys. Infections were more readily obtained by feeding on A. L. griseimembra than on A. nancymai. Transmission through sporozoites was obtained in an A. l. griseimembra monkey after a prepatent period of 24 days. PMID- 15357054 TI - Comparing parasite numbers between samples of hosts. AB - The comparison of parasite numbers or intensities between different samples of hosts is a common and important question in most parasitological studies. The main question is whether the values in one sample tend to be higher (or lower) than the values of the other sample. We argue that it is more appropriate to test a null hypothesis about the probability that an individual host from one sample has a higher value than individual hosts from a second sample rather than testing hypotheses about means or medians. We present a recently proposed statistical test especially designed to test hypotheses about that probability. This novel test is more appropriate than other statistical tests, such as Student's t-test, the Mann-Whitney U-test, or a bootstrap test based on Welch's t-statistic, regularly used by parasitologists. PMID- 15357055 TI - Helminth infracommunities of Rana vaillanti brocchi (Anura: Ranidae) in Los Tuxtlas, Veracruz, Mexico. AB - A total of 76 adult individuals of Rana vaillanti were collected in Laguna Escondida, Los Tuxtlas, Veracruz, Mexico, and their helminth infracommunity structure was determined. Among the 21 helminth taxa collected (10 digeneans, 8 nematodes, and 3 acanthocephalans), the digenean Langeronia macrocirra reached the highest prevalence (64.4%), mean abundance (6.6), and mean intensity (10.4), as well as the highest total number of individuals (499). Only 2 frogs were uninfected, the remainder harbored between 1 and 7 helminth species and 1-102 individuals; mean species richness and abundance were 3.49 +/- 0.22 and 16.1 +/- 16.3, respectively. Langeronia macrocirra dominated in 50.6% of the infracommunities, with relatively low Berger-Parker index values (0.56); for this reason, the evenness was high (0.70 +/- 0.31), and consequently, diversity values are the highest recorded to date in species of Rana. However, patterns of helminth infracommunity richness and diversity were similar to those previously observed in amphibians. This structure is attributed to the feeding habits (between 66.7 and 81% of helminth species parasitizing R. vaillanti enter using the food web dynamics) and low vagility (the remainder species infect by host penetration). PMID- 15357056 TI - Prospective study of swimmer's itch incidence and severity. AB - This epidemiologic study reports incidence, severity, and risk factors of swimmer's itch (cercarial dermatitis). Daily diaries about water exposures and swimmer's itch symptoms were completed by 40 riparian households at Douglas Lake, Michigan, for July 2000. Minutes spent in the water, minutes in shallow water, location, time of day, preventive action, age, and gender were recorded for all residents and guests. Incidence of swimmer's itch was 6.8 episodes per 100 water exposure days. Probability of an episode increased with more days of water use and at locations with onshore winds. Episode severity increased with more time in the water and at the same locations. Age and gender had no effect on incidence or severity. In sum, onset and severity of swimmer's itch are affected by how people interact with the lake, not by their demographic features. More studies of human incidence and severity are needed to convince public health agencies to address this problem at recreational lakes. Study designs that combine epidemiologic and biological data will simultaneously inform public health education and biological control programs. PMID- 15357057 TI - Nestedness in colonization-dominated systems: helminth infracommunities of Rana vaillanti Brocchi (Anura: Ranidae) in Los Tuxtlas, Veracruz, Mexico. AB - Colonization probabilities of parasite species often are determined by the habitat preference and vagility of host individuals. Although extinction-based interpretations have been investigated for nested subset patterns of parasite infracommunities, the low relative frequency of nestedness in colonization dominated systems makes the determination and interpretation of nested infracommunities of broad ecological importance. In these systems, ontogenetic shifts in habitat preference or diet of the host have the potential to produce nested subset patterns of parasite infracommunities. Helminth infracommunity structure was investigated for 76 Rana vaillanti individuals collected from Laguna Escondida, Los Tuxtlas, Veracruz, Mexico, in 1998. Pooled helminth infracommunities were significantly nested, as were penetrating and ingested helminth infracommunities when considered separately. Richness, diversity, and evenness of the helminth infracommunities were not correlated with host size, and did not differ between host sexes, suggesting that the structure of infracommunities simply is a product of the interaction between host individuals and their landscape mediated by individual differences in vagility. It is hypothesized that individual differences in recruitment can produce nested subset infracommunity patterns when the habitats or habitat preferences of hosts are themselves nested. PMID- 15357058 TI - Skewed Th1/Th2 immune response to Sarcoptes scabiei. AB - Scabies is a contagious skin disease of humans and many other species of mammals. Previous studies suggested that the balance between the Th1 and Th2 immune responses may influence the outcome of a scabies infestation in a sensitized host. Therefore, in this study, we examined the T-helper cell cytokine profiles of splenocytes and lymph node cells in BALB/c mice that were immunized with scabies extract (primary response), infested with scabies mites (primary response), or immunized and then infested (secondary response). Lymphocyte cytokine expression was analyzed by flow cytometry after staining for intracellular cytokines. Immunization with scabies extract induced production of interferon-gamma (IFNgamma) (Th1 response) by both spleen and lymph node cells. Mice that were infested with scabies increased production of interleukin-4 by lymph node cells and of IFNgamma by splenocytes. Mice that were first immunized and then infested with mites increased production of IFNgamma by both spleen and lymph node cells. However, this increased level of IFNgamma was only about half of that induced by immunization alone. These results suggest that live scabies mites produced something that inhibited IFNgamma production in the lymph nodes of scabies-immunized mice. Our data also indicate that lymphocytes in the spleen and lymph nodes can present different cytokine response profiles. PMID- 15357059 TI - Tickcidal effect of monoclonal antibodies against hemocytes, Om21, in an adult female tick, Ornithodoros moubata (Acari: Argasidae). AB - In the present study, monoclonal antibodies (mAbs) against adult Ornithodoros moubata hemocytes were established. Afterward, artificial feeding was performed to assess the tickcidal effect of fetal bovine serum meal containing each mAb. As a result, Om21 showed the strongest tickcidal effect on adult female O. moubata. The reactivity of various tick cells and organs, including the hemocyte, midgut, trachea, ovary, fat body, and muscle, to Om21 was then examined by an indirect immunofluorescent antibody test and by immunoelectron microscopy. Om21 reacted with not only hemocytes but also with fat body cells, epidermis, cuticle of the trachea, connective tissue of the muscle, and the basement membrane of the midgut, trachea, fat body, oocyte, and epidermis. These results suggest that Om21 passing through the midgut epithelium induced a tickcidal effect on hemocytes or various organs. However, the target of Om21 could not be identified in the present study. The antihemocyte mAb produced in this study, Om21, may be useful for the immunological control of ticks. PMID- 15357060 TI - Depletion of Schistosoma mansoni lung-stage schistosomula cholesterol by methyl beta-cyclodextrin dramatically increases specific antibody binding to surface membrane antigens. AB - Schistosoma mansoni lung-stage larvae appear to not bind antibodies from radiation vaccine or infection sera in the membrane immunofluorescence test. However, treatment of ex vivo lung-stage schistosomula with methyl-beta cyclodextrin, a hydrophobic oligosaccharide that specifically extracts cholesterol from plasma membranes, induced readily detectable binding of specific antibodies in a concentration- and time-dependent manner. Surface membrane antigen binding of specific antibodies was also conclusively demonstrated by quantitative absorption of anti-schistosome sera with intact ex vivo larvae. These data together suggest that confinement of lung-stage schistosomula surface membrane antigens in cholesterol-rich sites allows only monovalent antibody binding, which can be detected by absorption and not by direct serology. PMID- 15357061 TI - Schistosoma mansoni: antigen-presenting cells emigrating from skin exposed to attenuated cercariae activate lymphoid cells and transfer protection in C57B1/6 mice. AB - C57B1/6 mice develop significant levels of protection to a challenge infection after percutaneous exposure to irradiated Schistosoma mansoni cercariae. Although some circumstantial evidence has suggested that antigen-presenting cells (APCs) within the skin play a role in priming anti-schistosomula effector mechanisms, no direct evidence has been presented. In this study, we describe efforts to directly test whether skin-resident APCs exposed to irradiated cercariae are capable of mediating responses consistent with previously proposed mechanisms associated with delayed-type hypersensitivity reactions. We demonstrate that a population of APCs emigrates from the skin after percutaneous vaccination and that these cells are able to induce proliferation of S. mansoni-specific lymphocytes. We describe our experiments conducted to confirm that proliferation is dependent on major histocompatibility complex (MHC) Class-II interactions and cell-to-cell contact between APCs and lymphocytes. Immunohistological staining of emigrating cells revealed a population of large MHC Class-II+ cells with a morphology characteristic of mature dendritic cells. On recovery and adoptive transfer into naive mice, these cells demonstrated the ability to mediate protection to a challenge infection at levels similar to those in percutaneously vaccinated controls. This confirms that cutaneous APCs can initiate anti schistosomula effector mechanisms in C57B1/6 mice after percutaneous vaccination. PMID- 15357062 TI - Differential effects of interferon-gamma on production of trypanosome-derived lymphocyte-triggering factor by Trypanosoma brucei gambiense and Trypanosoma brucei brucei. AB - Trypanosome-derived lymphocyte-triggering factor (TLTF) produced by Trypanosoma brucei brucei stimulates production of interferon-gamma (IFN-gamma) by CD8+ T cells, and it is reported that, in turn, IFN-gamma stimulates proliferation of T. b. brucei. We studied the role of TLTF in trypanosome proliferation using the Wellcome strain (WS) of Trypanosoma brucei gambiense and the ILtat 1.4 strain (IL) of T. b. brucei. Increase in the number of WS in infected rats is more rapid than IL and corresponds with comparatively higher levels of IFN-gamma. Production of IFN-gamma, as measured by protein and messenger RNA (mRNA) levels, was maintained by splenocytes from WS-infected rats, whereas levels decreased in IL infected rats, accompanied by prolongation of infection. Expression of TLTF mRNA by in vitro-cultured WS was promoted in a dose-dependent fashion by addition of recombinant rat IFN-gamma at all concentrations tested. The addition of lower concentrations of IFN-gamma to cultured IL increased expression of TLTF mRNA, whereas, in contrast to WS, addition of 100 and 1,000 U/ml IFN-gamma decreased expression of TLTF by IL. These results show that unlike WS, elevated IFN-gamma concentrations lead to decreased TLTF production by IL. It is believed that decreased TLTF production in IL-infected rats leads to lowered IFN-gamma production, thereby slowing IL proliferation. PMID- 15357064 TI - Kinetics of Echinostoma caproni (Trematoda: Echinostomatidae) antigens in feces and serum of experimentally infected hamsters and rats. AB - This study reports on the kinetics of antibody production to Echinostoma caproni and the dynamics of antigens in feces and sera in 2 experimental hosts (hamsters and rats) that display different degrees of susceptibility with this echinostome. Echinostoma caproni produced chronic infections in hamsters, whereas rats lost the infection at 49-56 days postinfection (DPI). Hamsters developed higher antibody responses than rats, probably in relation to different intestinal absorptions of worm antigens in each host species. The levels of coproantigens were indicative of the course of infection in each host. Positive coproantigen levels were detected at 1-2 DPI in both hosts, and the values remained positive until the end of the experiment in hamsters; in rats, the coproantigen levels reverted to negative values, coinciding with the loss of infection. High levels of circulating antigens were detected in hamsters from 21 DPI to the end of the study. In contrast, low levels of E. caproni seroantigens were detected in rats only. These observations may reflect the differences in local inflammatory responses induced by E. caproni in each host species. PMID- 15357063 TI - A recombinant antigen recognized by Fasciola hepatica-infected hosts. AB - This work reports the detection of specific immunoglobulins (Ig) against rFh8, a recombinant Fasciola hepatica adult worm excretion-secretion antigen, in sera from experimentally (rabbit, Wistar rat, cattle, and sheep), or naturally (human) infected hosts. In the case of laboratory experimental models the study revealed significant differences between rabbits, which recognized the recombinant antigen all along the infection, and Wistar rats, which showed high anti-rFh8 Ig levels only for a short period of the infection. Available sera from experimentally infected cattle and sheep, as well as sera from naturally F. hepatica-infected humans, also contained significant levels of Ig against rFh8, suggesting that Fh8 was produced by F. hepatica at a very early stage of infection in all hosts so far analyzed and that the rFh8 antigen could be used as a tool for the diagnosis of F. hepatica infections. PMID- 15357065 TI - Molecular identification of symbionts from the pulmonate snail Biomphalaria glabrata in Brazil. AB - The icthyosporean, Capsaspora owczarzaki, a known predator of Schistosoma mansoni sporocysts in vitro, is more prevalent in laboratory-reared strains of the intermediate snail host, Biomphalaria glabrata resistant to S. mansoni, than from the susceptible M line strain. We examined whether B. glabrata resistant to the NIH-PR-1 strain of S. mansoni from 2 regions in Brazil were also host to C. owczarzaki. Symbiont presence was examined using hemolymph culturing and nested polymerase chain reaction of snail genomic DNA with primers designed to specifically amplify sequences from relatives of the Icthyosporea. All B. glabrata of the resistant Salvador strain from the laboratory of Dr. Lobato Paraense in Rio de Janeiro, Brazil (n = 46) tested negative for symbionts. Three of 18 semiresistant 10-R2 B. glabrata from the laboratory of Dr. Barbosa in Recife, Brazil tested positive for C. owczarzaki. Another icthyosporean, Anurofeca sp., was identified from 1, 10-R2 snail and from 2 of 12 field collected B. glabrata from Praia do Forte Orange, Ilha de Itamaraca. Snails from 2 other sites, Hotel Colibri, Pontezinha and Praia do Sossego, Ilha de Itamaraca, were negative for Anurofeca. Two genera of ciliates were also identified. Paruroleptus sp. was found in 4, 10-R2 snails and Trichodina sp. was identified in 2 field-collected snails from Praia do Forte Orange and Praia do Sossego. PMID- 15357066 TI - Resistance of early midgut stages of natural Plasmodium falciparum parasites to high temperatures in experimentally infected Anopheles gambiae (Diptera: Culicidae). AB - We studied the effects of high temperature, 30 and 32 versus 27 C on early Plasmodium falciparum development in Anopheles gambiae experimentally infected with gametocytes from 30 volunteers with mean density of 264.1 gametocytes/microl blood (range: 16-1,536/microl). From several batches of mosquitoes, fed by membrane feeding, midguts of individual mosquitoes were dissected at 24 hr for ookinete enumeration and at 7 days to quantify oocysts. There were temperature related differences in mean ookinete intensity per mosquito midgut, with 9.71 +/- 1.6 at 27 C, 9.85 +/- 2.32 at 30 C, and 3.89 +/- 0.81 at 32 C. The prevalence of oocyst infection decreased with an increase in temperatures from 15.9 to 8.5 to 6.4% at 27, 30, and 32 C, respectively. The average oocyst intensities for the infected mosquitoes increased with temperatures from 2.9 at 27 C to 3.5 at 30 C, and to 3.3 at 32 C. However, the success of infections was reduced at 30 and 32 C, and resulted in greater losses during consecutive inter-stage parasite development. The most significant impact of high temperatures occurred at the transition between macrogametocytes and ookinetes, whereas the transition between ookinetes and oocysts apparently was not affected. In contrast to other reports, exposure of mosquitoes infected with natural parasites to high temperatures did not eliminate preoocyst stages, as has been observed from laboratory studies using the NF-54 strain of P. falciparum. This observation of parasite resistance to high temperatures is consistent with the natural situation in tropical environments where perennial malaria transmission occurs during hot dry seasons. PMID- 15357067 TI - Cryptosporidium suis n. sp. (Apicomplexa: Cryptosporidiidae) in pigs (Sus scrofa). AB - Molecular and biological characteristics of a new species of Cryptosporidium from the feces of pigs (Sus scrofa) is described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum; they are passed fully sporulated, lack sporocysts, and measure 4.9-4.4 microm (mean = 4.6 microm) x 4.0 4.3 microm (mean = 4.2 microm); length to width ratio 1.1 (n = 50). Cryptosporidium suis is not transmissible to nude mice and is poorly infectious for cattle. Molecular and phylogenetic analyses at the 18S ribosomal RNA, heat shock protein 70, and actin gene loci demonstrate C. suis to be genetically distinct from all known species and genotypes of Cryptosporidium, and thus is named as Cryptosporidium suis. PMID- 15357068 TI - Larva migrans by Baylisascaris transfuga: fatal neurological diseases in Mongolian jirds, but not in mice. AB - Raccoon roundworms (Baylisascaris procyonis) and other Baylisascaris species cause patent or latent larva migrans (LM) in a variety of mammals and birds, including humans. It is not clear whether LM by Baylisascaris transfuga, roundworms of bears, is associated with clinical neurological disorders. To clarify this issue, ICR and BALB/c mice as well as Mongolian jirds (Meriones unguiculatus) were orally inoculated with 2,000-5,000 embryonated eggs of B. transfuga. In mice, the ascarid caused symptomatic LM of limited extent and duration, whereas the infection was fatal in jirds; i.e., they exhibited general signs such as severe depression and emaciation on days 8-11 postinfection (PI) and died, or they developed progressive and fatal neurological disorders after day 14 PI. Histological examination showed B. transfuga larvae in the brain of all mice and jirds examined, and the larvae collected from them developed to a size comparable with that of B. procyonis. There existed, however, critical differences in host reactions against larvae localized in the brain of mice and jirds; B. transfuga larvae found in mice were surrounded by granulomatous reactions and immobilized, whereas larvae found in jirds were free from any host reaction and mobile, causing extensive malacia. PMID- 15357069 TI - Depletion of natural killer cells does not result in neurologic disease due to Sarcocystis neurona in mice with severe combined immunodeficiency. AB - Sarcocystis neurona is an apicomplexan parasite that is the primary etiologic agent of equine protozoal myeloencephalitis in horses. Protective immune responses in horses have not been determined, but interferon-gamma (IFN-gamma) is considered critical for protection from neurologic disease in mice. The role of adaptive and innate immune responses in control of parasites was explored by infecting BALB/c, IFN-gamma knockout (GKO), and severe combined immune deficient (SCID) mice with S. neurona (10(4) sporocysts/mouse). Immune competent BALB/c mice eliminated parasites within 30 days, with no sign of neurologic disease, whereas GKO mice developed fulminant neurologic disease. In contrast, SCID mice remained healthy throughout the experimental period despite the persistence of parasite at low levels in some mice. Treatment with anti-IFN-gamma antibody resulted in neurologic disease in infected SCID mice. Although SCID mice lack adaptive immune responses, they have natural killer (NK) cells capable of producing significant quantities of IFN-gamma. Therefore, SCID mice were infected with sporocysts of S. neurona and treated with anti-asialo GM1. Depletion of NK cells, confirmed by flow cytometry, did not result in neurologic disease in SCID mice. These results indicate that IFN-gamma mediates protection from neurologic disease in SCID mice. Protective levels of IFN-gamma may originate from a low number of nondepleted NK cells or from a non-T cell, non-NK cell population. PMID- 15357070 TI - A new species of Oceanobdella (Hirudinida: Piscicolidae) from the plain sculpin, Myoxocephalus jaok, from Bristol Bay, Alaska. AB - Oceanobdella khani n. sp. is described from the plain sculpin, Myoxocephalus jaok, from Bristol Bay, Alaska, in the eastern Bering Sea. Prevalence was never greater than 10% in any single collection; maximum intensity was 12 leeches per host, but most fish had 5 or fewer leeches. Oceanobdella khani possesses generic characters of small oral sucker, 5 pairs of testisacs, unremarkable terminal male reproductive organs, coelomic system lacking pulsatile vesicles, and 3 pairs of eyes on the oral sucker-trachelosome. Oceanobdella khani is distinguished from other species in the genus by solid black pigmentation on the urosome, clitellum, trachelosome, and most of the oral sucker except for an unpigmented margin. The pigmentation of the caudal sucker is highly variable, ranging from overall mottled blackish gray to completely unpigmented. The caudal sucker lacks ocelli. Intestinal ceca are large, crop ceca are directed anteriorly, and postceca are separate for their entire length. PMID- 15357071 TI - Description of Brachylecithum mackoi n. sp. (Digenea: Dicrocoeliidae) from the European hedgehog, Erinaceus europaeus (Insectivora: Erinaceidae). AB - A new hepatic dicrocoeliid species, Brachylecithum mackoi n. sp. (Digenea, Dicrocoeliidae), is described from the European hedgehog, Erinaceus europaeus (L.) (Insectivora, Erinaceidae). An infected host was found in the Mediterranean island of Elba (Italy), and more than 60 individuals were isolated from the biliary ducts. The holotype and 55 paratypes were examined. Brachylecithum mackoi n. sp. differs from congeneric species found in mammal hosts by having well developed lappets in the ventral sucker, a sloping uterus between anterior testis and acetabulum, no overlap between vitellaria, and metrical features in the body size, sucker diameters, cirrus sac, and size of eggs. The only other Brachylecithum species of erinaceids in Europe and Africa, Brachylecithum aetechini Dollfus, 1951, differs from the new species in the above-mentioned morphological characters, greater dimensions of the body, and oral sucker, pharynx, cirrus sac, and egg dimensions. The presence of B. mackoi n. sp. in Elba Island is discussed in the light of apparent host specificity of erinaceid dicrocoeliids and geographical distribution of Palearctic and Ethiopian Erinaceidae. PMID- 15357073 TI - A new species of Pelecitus (Filarioidea: Onchocercidae) from the endangered Tehuantepec jackrabbit Lepus flavigularis. AB - Pelecitus meridionaleporinus n. sp. from the Tehuantepec jackrabbit is described. The new species differs from Pelecitus helicinus (Molin, 1860) in having delicate transverse striations, a salient vulva, and a readily apparent preesophageal ring; P. helicinus has teardrop cells around the vulva, which are lacking in the species presently described. The new species is different from Pelecitus scapiceps (Leidy, 1886) in having the vulva anterior to the esophageal-intestinal junction and wider lateral alae. Pelecitus scapiceps is found in the tarsal bursa of the hind feet of lagomorphs, whereas P. helicinus is found around tendons of legs and feet of birds. Pelecitus meridionaleporinus n. sp. occurs in the subcutaneous tissue at the base of both ears. This is the second species in Pelecitus Railliet and Henry, 1910 that occurs in New World lagomorphs, and the third found infecting mammals. PMID- 15357072 TI - A new PCR assay for simultaneous studies of Leucocytozoon, Plasmodium, and Haemoproteus from avian blood. AB - Many bird species host several lineages of apicomplexan blood parasites (Protista spp., Haemosporida spp.), some of which are shared across different host species. To understand such complex systems, it is essential to consider the fact that different lineages, species, and families of parasites can occur in the same population, as well as in the same individual bird, and that these parasites may compete or interact with each other. In this study, we present a new polymerase chain reaction (PCR) protocol that, for the first time, enables simultaneous typing of species from the 3 most common avian blood parasite genera (Haemoproteus, Plasmodium, and Leucocytozoon). By combining the high detection rate of a nested PCR with another PCR step to separate species of Plasmodium and Haemoproteus from Leucocytozoon, this procedure provides an easy, rapid, and accurate method to separate and investigate these parasites within a blood sample. We have applied this method to bird species with known infections of Leucocytozoon spp., Plasmodium spp., and Haemoproteus spp. To obtain a higher number of parasite lineages and to test the repeatability of the method, we also applied it to blood samples from bluethroats (Luscinia svecica), for which we had no prior knowledge regarding the blood parasite infections. Although only a small number of different bird species were investigated (6 passerine species), we found 22 different parasite species lineages (4 Haemoproteus, 8 Plasmodium, and 10 Leucocytozoon). PMID- 15357074 TI - Cucullanus bonaerensis n. sp. (Nematoda: Cucullanidae) parasitizing Urophycis brasiliensis (Pisces: Phycidae) from Argentinean waters. AB - A total of 8 specimens of Urophycis brasiliensis (Kaup, 1858) from waters off Mar del Plata, Argentina (38 degrees 08'S, 57 degrees 32'W), were examined for parasitic nematodes. A new nematode species, Cucullanus bonaerensis n. sp., is described (prevalence 50%, mean intensity 15.5). The new species differs from its congeners in the distribution pattern of caudal papillae (particularly fourth pair) and phasmids, length of the body and spicules, and by the position of the excretory pore (in the posterior third of esophagus). Species of Cucullanus, reported previously as C. heterochrous Rudolphi, 1812, and Cucullanus sp. from U. brasiliensis from Puerto Quequen, Argentina (38 degrees 37'S, 58 degrees 53'W), are considered as members of the new species. PMID- 15357075 TI - Philometroides buirnurensis n. sp. (Nematoda: Philometridae) from cyprinid fishes in the Buir Nur Lake on the border of China and Mongolia. AB - A new species of philometrid nematode (Nematoda: Philometridae), Philometroides buirnurensis sp. n., is described and illustrated on the basis of female specimens from subcutaneous tissues of the inner surface of the abdominal cavity and the bile duct of cyprinid fishes Leucisuus waleckii (Dybowski), 1869 (type host), Pseudaspius leptocephalus (Pallas), 1776 and Hemiibarbus labeo (Pallas), 1776, from the Buir Nur Lake, a boundary lake between China and Mongolia, on southwest of the Hulun Buir League in the Inner Mongolia Autonomous Region of China. The new species distinctly differs from its congeners mainly in that the posterior half of its body is tapered and conspicuously narrower than the anterior region and that both the head and tail ends are rounded. The cuticular bosses are distributed extensively at both ends but poorly on the middle part of the body. The most peculiar feature of the bosses situated at both ends was their lobe- or rodlike shape and their transverse orientation. Four pairs of submedian outer cephalic papillae and 1 pair of caudal papilla are all relatively large. The new species differs from the most similar congener, Philometroides ganzhouensis Yu, 1998, in having 4 inner cephalic papillae and in the presence of 2 caudal papillae in mature specimens. PMID- 15357077 TI - Similascarophis n. gen. n. spp. (Nematoda: Cystidicolidae) parasitizing marine fishes off the Chilean coast. AB - Similascarophis (Cystidicolidae) n. gen. is proposed. In the mouth of specimens of this genus, submedial labia are absent and pseudolabia do not have any part projecting toward the central oral opening. These nematodes were obtained from the alimentary tract of 7 marine fish species along the coast of Chile: Bovichthys chilensis Regan, Eleginops maclovinus (Cuvier), Pinguipes chilensis (Valenciennes), Cilus gilberti (Abbott), Cheilodactylus variegatus Valenciennes, Girella laevifrons (Tschudi), and Graus nigra Philippi. Morphology and morphometry are compared between 2 new Similascarophis species: Similascarophis maulensis n. sp. and S. chilensis n. sp., which differ in the presence of sublabia and in the length of the glandular esophagus and left spicule. We also recorded Similascarophis sp. in 2 other host species, which showed some distinct proportional measurements, although these differences were not sufficiently clear to identify them as a new species. PMID- 15357076 TI - Identification of European diplozoids (Monogenea, Diplozoinae) by restriction digestion of the ribosomal RNA internal transcribed spacer. AB - The second internal transcribed spacer (ITS2) of the ribosomal RNA genes of Diplozoon paradoxum and Paradiplozoon nagibinae were amplified and sequenced. The polymerase chain reaction product of D. paradoxum was bigger (840 bp) than that of P. nagibinae (820 bp). There was no intraspecific variability recorded in sequences from either species. Sequence comparisons and ITS2 restriction fragment length polymorphism (RFLP) pattern of 8 European diplozoid species aimed to resolve their identification and amend the previous studies. RFLP was used to distinguish the 2 species from each other and from P. bliccae, P. homoion, P. megan, P. pavlovskii, P. sapae, and Eudiplozoon nipponicum, using restriction enzymes AluI, HaeIII, HinfI, RsaI, and SphI. The criteria for morphological identification of 8 European diplozoids are also included, with the main morphological characters of clamps, trapeze spur, and anterior joining sclerites of 8 diplozoid species being illustrated. Combination of the shape and comparison of length of the trapeze spur and anterior joining sclerites could lead to accurate identification of diplozoid species. PMID- 15357078 TI - Comparison of synthetic tyvelose antigen with excretory-secretory antigen for the detection of trichinellosis in swine using enzyme-linked immunosorbent assay. AB - Two enzyme-linked immunosorbent assay (ELISA) systems, one using natural excretory-secretory (ES) antigens and the other a synthetic glycan antigen (3,6 dideoxy-D-arabinohexose [tyvelose, TY]), were evaluated for the serological diagnosis of trichinellosis in swine. Sensitivity was estimated using samples (n = 113) collected 3-21 wk PI from 15 experimentally infected pigs, and specificity was estimated using samples (n = 397) from a population of Trichinella spp.-free pigs. Results were analyzed using 2 cutoff values recommended in international guidelines (Office Internationale des Epizooties [OIE]) and by the optimal cutoff level as determined by receiver-operator characteristic (ROC) analysis. The ROC optimized TY-ELISA consistently performed better than all other combinations. None of the combinations of test and cut-off detected infected pigs sooner than 35 days; however, the ROC-optimized TY-ELISA identified 8 of 15 pigs earlier than the ES-ELISA and detected 2 pigs missed by all other tests. At 49 days PI the sensitivity and specificity of the ROC-optimized TY-ELISA were 94.3 and 96.7%, respectively, as compared with the ROC-optimized ES-ELISA at 84.9 and 96.0%, respectively. The ROC-optimized TY-ELISA was 100% specific at OIE-recommended cut offs. This study indicates that the TY-ELISA is as good or better than the ES ELISA for the detection of trichinellosis in swine. PMID- 15357079 TI - Lethal effects of freezing Echinococcus multilocularis eggs at ultralow temperatures. AB - Methods for killing Echinococcus multilocularis eggs within stool or intestinal samples, without damaging the diagnostic value of the sample, would significantly reduce the risk of animal health providers acquiring alveolar hydatid disease. The first objective of this study was to determine whether E. multilocularis eggs located in fox intestines can survive storage at -70 C for at least 4 days. Results showed that none of 72,000 E. multilocularis eggs remained infectious to defined strains of mice under these conditions, yet, similar eggs recovered from nonfrozen carcasses stored at 4 C for the same time period were viable. The structural identities of adult worms and eggs were not significantly altered by the freezing and thawing processes. These results indicate that ultracold temperatures can be used to kill or inactivate E. multilocularis eggs, making them safe to handle when diagnosing this parasite in definitive hosts. The second objective of this study was to determine whether E. multilocularis eggs could survive freezing to -70 C if commonly used cryopreservation protocols were used. The use of the cryoprotectant solution, 5% dimethyl sulfoxide-35% saline-60% lamb serum, with a -1 C/min freezing rate was unable to prevent the eggs from being killed by freezing to -70 C. Rapid cooling by plunge freezing into liquid nitrogen was also lethal to E. multilocularis eggs. Only a few of the many potential cryopreservation protocols were tested in this study, so it is not yet possible to completely rule out the possibility of preserving these eggs at ultralow temperatures, but it does indicate that temperatures below -70 C are lethal to eggs even under favorable storage conditions. PMID- 15357080 TI - An ultrasensitive capture ELISA for detection of Fasciola hepatica coproantigens in sheep and cattle using a new monoclonal antibody (MM3). AB - A capture enzyme-linked immunosorbent assay (ELISA) using a new monoclonal antibody (mAb MM3) is reported for the detection of Fasciola hepatica excretory secretory antigens (ESAs) in feces of infected hosts. The mAb MM3 was produced by immunization of mice with a 7- to 40-kDa purified and O-deglycosylated fraction of F. hepatica ESAs, which has previously been shown to be specific for the parasite. The specificity and sensitivity of the MM3 capture ELISA were assessed using feces from sheep and cattle. Sheep feces were obtained from a fluke-free herd (with most animals harboring other nematodes and cestodes), from lambs experimentally infected with 5-40 F. hepatica metacercariae and in some cases treated with triclabendazole at 14 wk postinfection (PI), and from uninfected control lambs. Cattle feces were collected at the slaughterhouse from adult cows naturally infected with known numbers of flukes (from 1 to 154) or free of F. hepatica infection (though in most cases harboring other helminths). The MM3 capture ELISA assay had detection limits of 0.3 (sheep) and 0.6 (cattle) ng of F. hepatica ESA per milliliter of fecal supernatant. The assay detected 100% of sheep with 1 fluke, 100% of cattle with 2 flukes, and 2 of 7 cattle with 1 fluke. The false-negative animals (5/7) were probably not detected because the F. hepatica individuals in these animals were immature (5-11 mm in length). As expected, coproantigen concentration correlated positively (r = 0.889; P < 0.001) with parasite burden and negatively (r = 0.712; P < 0.01) with the time after infection at which coproantigen was first detected. Nevertheless, even in animals with low fluke burdens (1-36 parasites), the first detection of F. hepatica specific coproantigens by the MM3 capture ELISA preceded the first detection in egg count by 1-5 wk. In all sheep that were experimentally infected and then untreated, coproantigen remained detectable until at least 18 wk PI, whereas in sheep that were experimentally infected and then flukicide treated, coproantigen became undetectable from 1 to 3 wk after treatment. None of the fecal samples from sheep or cattle negative for fascioliasis but naturally infected with other parasites including Dicroelium dendriticum showed reactivity in the MM3 capture ELISA. These results indicate that this assay is a reliable and ultrasensitive method for detecting subnanogram amounts of F. hepatica antigens in feces from sheep and cattle, facilitating early diagnosis. PMID- 15357081 TI - Clinical features, epidemiology, and efficacy and safety of intralesional antimony treatment of cutaneous leishmaniasis: recent experience in Turkey. AB - A total of 1,030 patients, 40.2% men and 59.8% women, identified during the period of October 1998 to November 2002 as having cutaneous leishmaniasis (CL), were studied; 1,431 lesions were identified in the 1,030 patients. One lesion was present in 80.7% of the patients. The size of the lesions (longest axis) was 13.6 mm (standard, 12.1 mm; range 3-150 mm). Most of the lesions were of the papular type (51.2%), although several atypical clinical presentations of CL were observed. The duration of the disease ranged between 1 and 72 mo (mean duration, 10.8 mo). The clinical suspicion of CL was confirmed by the observation of amastigotes on lesion tissue samples stained by Giemsa. The test was positive in 851 of 1,030 patients (82.6%). Intralesional meglumine antimonate solution (85 mg Sb/ml, 0.2-1 ml, depending on the size of the lesion) weekly until complete cure or up to 20 wk was used for first-line therapy of 890 patients (86.4%). We found that this regimen of intralesional Sb has an efficacy of 97.2% with a low relapse rate of 3.9% and no serious adverse side effects. PMID- 15357082 TI - Uncinaria hamiltoni (Nematoda: Ancylostomatidae) in South American sea lions, Otaria flavescens, from northern Patagonia, Argentina. AB - Thirty-one South American sea lion pups (Otaria flavescens) found dead in Punta Leon, Argentina, during the summer of 2002, were examined for hookworms (Uncinaria hamiltoni). Parasite parameters were analyzed in 2 locations of the rookery, i.e., a traditional, well-structured breeding area and an expanding area with juveniles and a lax social structure. Prevalence of hookworms was 50% in both localities, and no difference was observed in prevalence between pup sexes (P > 0.05). Hookworms were concentrated in the small intestine. Transmammary transmission is assumed because only adult hookworms were found in the pups. The mean intensity of hookworms per pup was 135; the mean intensity in females (92.78) was significantly different (P < 0.05) from that of males (230.25). No difference (P > 0.05) in intensity was found between the 2 breeding areas, although prevalence was higher in the traditional breeding area than in the other area. Location was the only factor affecting hookworm prevalence (P log-linear model: 0.9552; chi2: 1.5629). No apparent trend between body condition and intensity of hookworms was observed. PMID- 15357083 TI - First isolation of Neospora caninum from an aborted bovine fetus in Spain. AB - Neospora caninum was isolated from the brain of a 6-mo-old aborted bovine fetus from Galicia, Spain. The fetal brain homogenate was inoculated intraperitoneally into cortisonized mice. The peritoneal exudate from the infected mice, along with mouse sarcoma cells (Tg180), was inoculated into a second group of mice, and parasites were harvested from the peritoneal exudate. The parasites were adapted to in vitro growth in Vero monolayers. The tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti Toxoplasma gondii antibodies on indirect fluorescent antibody test. The tachyzoites were lethal to interferon gamma gene knock out (KO) mice and could be identified immunohistochemically in the tissues. The identity of the parasite was also confirmed by polymerase chain reaction amplification of N. caninum-specific fragments. The sequences of the amplified gene 5 fragments (GenBank AY494944) were found to be identical to that of an Austrian isolate of N. caninum but not to that of NC-1. This is the first isolation of viable N. caninum from Spain. PMID- 15357084 TI - First report of Neospora caninum infection in adult alpacas (Vicugna pacos) and llamas (Lama glama). AB - Neospora caninum is a cyst-forming coccidian that mainly affects bovines, although Neospora infection has also been described in other domestic and wild ruminant species. Serum samples from 78 alpacas (Vicugna pacos) and 73 llamas (Lama glama) at a unique dilution of 1:50 tested by indirect fluorescent antibody test (IFAT) were further analyzed serologically by IFAT and Western blot in both ruminant species to avoid cross-reactions with closely related coccidian parasites and to confirm the existence of N. caninum-specific antibodies. IFAT titers ranging between 1:50 and 1:800 were found. When using Western blot, N. caninum tachyzoite-specific immunodominant antigens with apparent molecular weights of 17-18, 34-35, 37, and 60-62 kDa were also recognized, although some sera with 1:50 IFAT titers proved not to have N. caninum-specific antibodies. As expected, higher IFAT titers were associated with higher anti-N. caninum reactivity in Western blot. This report documents for the first time the presence of N. caninum infection in adult alpacas and llamas from Peru. PMID- 15357085 TI - Additional observations on the sporozoite transmission of Plasmodium knowlesi to monkeys. AB - Saimiri boliviensis monkeys were infected by the intravenous injection of 50 sporozoites of the H strain of Plasmodium knowlesi dissected from the salivary glands of Anopheles dirus mosquitoes; prepatent periods were 11, 12, 13, 13, 13, and 16 days. Sporozoites of P. knowlesi stored frozen for 7 days, 53 days, 20 mo, 7 yr and 7 mo, and 11 yr and 5 mo induced infections in Macaca mulatta monkeys with prepatent periods of 7, 6, 8, 10, and 7 days, respectively. After frozen storage for 11 yr and 5 mo, infections were induced in S. boliviensis with prepatent periods of 10-13 days. PMID- 15357086 TI - Beta-tubulin complementary DNA sequence variations observed between cyathostomins from benzimidazole-susceptible and -resistant populations. AB - The molecular mechanism of benzimidazole (BZ) resistance in cyathostomins of horses is still unclear. Previous studies revealed that the TTC or TAC polymorphism in codon 200 of the beta-tubulin isotype 1 gene is not as strictly correlated with BZ resistance as in trichostrongyles in sheep. To identify further sites of polymorphism within the beta-tubulin gene related to BZ resistance, complete complementary DNAs (cDNAs) encoding beta-tubulin of adult worms of Cylicocyclus nassatus, Cyathostomum pateratum, Cyathostomum coronatum, Cyathostomum catinatum, Cylicostephanus longibursatus, and Cylicostephanus goldi of a BZ-resistant cyathostomin population were characterized using specific primers. The cDNA sequence of each species spans 1,429 bp, encoding a protein of 448 amino acids. The interspecific identities are 95.2-99.6% at the nucleotide and 98.7-100.0% at the peptide level. The comparison of the amino acid sequences of individuals isolated from the BZ-resistant cyathostomin population with those from individuals of Cc. nassatus, Cy. coronatum, Cy. pateratum, and Cy. catinatum of a BZ-susceptible one showed differing amino acids in 11 positions. The commonness of a phenylalanine to tyrosine mutation at position 167 in all the 6 cyathostomin species isolated from a BZ-resistant population suggests its involvement in the molecular mechanism in BZ resistance. PMID- 15357087 TI - Seroprevalence of Toxoplasma gondii in wild toque macaques (Macaca sinica) at Polonnaruwa, Sri Lanka. AB - From a natural population that inhabits the dry evergreen forest at Polonnaruwa, serum samples of 170 toque macaques were examined for antibodies to Toxoplasma gondii by the modified agglutination test. Of these, 21 (12%) were found with titers of 1:16 in 9, 1:32 in 9, 1:256 in 1, 1:1,024 in 1, and 1:4,096 in 1. There was no evidence of maternal transmission of antibodies or congenital toxoplasmosis. None of the infected macaques died within 1 yr after sampling. Toxoplasma gondii infection was closely linked to human environments where domestic cats were common. Macaques having frequent contact with human settlements showed a significantly greater (P < 0.0001) prevalence (19% infected) than macaques restricted to forest habitat, none of which was infected. Although infection with T. gondii has been noted in several species of Asian primates, this is the first report of T. gondii antibodies in toque macaques (Macaca sinica) that are endemic to the island of Sri Lanka. PMID- 15357088 TI - Assessment of Sarcocystis neurona sporocyst viability and differentiation between viable and nonviable sporocysts using propidium iodide stain. AB - Sarcocystis neurona has become recognized as the major causative agent of equine protozoal myeloencephalitis (EPM) in the Americas. At least 3 pathogenic species of Sarcocystis, including S. neurona, can be isolated from opossums. Methods are needed to ascertain whether these isolates are viable and capable of causing infections. In this study, the nuclear stain propidium iodide (PI) was used to differentiate between live (viable) and heat-killed (nonviable) S. neurona sporocysts. PI was excluded by live sporocysts but penetrated compromised sporocyst membrane and stained sporozoite nuclei of dead sporocysts. After live and dead sporocysts were mixed at various ratios, the number of unstained sporocysts detected after the staining procedure correlated significantly (r2 = 0.9978) with the expected numbers of live sporocysts. Sporocyst mixtures were also assayed for in vitro excystation and development in tissue cultures. The correlation between the percentage of plaques formed in tissue cultures and the percentage of expected infectious (live) sporocysts in each mixture was r2 = 0.6712. By analysis of variance, no statistically significant difference was measured between the percentage of viable sporocysts and the percentage of infectious sporocysts (P = 0.3902) in each mixture. In addition, there was evidence of a relation between PI impermeability of sporocysts and animal infectivity. These results suggest that the PI dye-exclusion technique can be a useful tool in identifying viability and potential infectivity of S. neurona sporocysts and in differentiating between viable and nonviable sporocysts. PMID- 15357090 TI - A rapid method for counting nucleated erythrocytes on stained blood smears by digital image analysis. AB - Measures of parasitemia by intraerythrocytic hematozoan parasites are normally expressed as the number of infected erythrocytes per n erythrocytes and are notoriously tedious and time consuming to measure. We describe a protocol for generating rapid counts of nucleated erythrocytes from digital micrographs of thin blood smears that can be used to estimate intensity of hematozoan infections in nonmammalian vertebrate hosts. This method takes advantage of the bold contrast and relatively uniform size and morphology of erythrocyte nuclei on Giemsa-stained blood smears and uses ImageJ, a java-based image analysis program developed at the U.S. National Institutes of Health and available on the internet, to recognize and count these nuclei. This technique makes feasible rapid and accurate counts of total erythrocytes in large numbers of microscope fields, which can be used in the calculation of peripheral parasitemias in low intensity infections. PMID- 15357089 TI - A novel Fasciola hepatica saposinlike recombinant protein with immunoprophylactic potential. AB - A member of the Fasciola hepatica saposinlike/NK-lysin protein family with lytic activity on human peripheral blood mononuclear cells and erythrocytes was recently described. The current study was designed to test the immunoprophylactic potential of this protein termed FhSAP-2 against infection with F. hepatica in rabbits. Two doses of 50 microg of recombinant FhSAP-2 (rFhSAP-2) emulsified in TiterMax were injected subcutaneously on the dorsal surface of 4 rabbits at 2-wk intervals. Four weeks after the second immunization, the rabbits were infected orally with 25 F. hepatica metacercariae. Four non-immunized-infected rabbits were used as controls. An enzyme-linked immunosorbent assay revealed high levels of antibodies to both rFhSAP-2 and F. hepatica excretory-secretory antigens by 2 wk after the first immunization, which were always significantly higher in immunized-infected rabbits than in control-infected rabbits. On the completion of the trial, vaccinated rabbits had 81.2% less flukes than controls. Moreover, F. hepatica egg counts in feces, as well as in bile collected from the gall bladders from vaccinated animals, were lower, 83.8 and 73%, respectively, compared with controls. The vaccinated rabbits also had significantly lower amounts of parasite antigen in stool and bile samples than controls. Last, evaluation of macroscopic liver lesions revealed that the rabbits vaccinated with rFhSAP-2 had milder lesions than the infected-control rabbits. These findings support the hypothesis that this novel rFhSAP-2 protein has immunoprophylactic potential against fascioliasis in rabbits including antifecundity and antipathology effects. This is the first report on experimental vaccination of rabbits against F. hepatica with a purified, defined, recombinant protein related to a member of the saposinlike protein family. PMID- 15357091 TI - Prevalence of agglutinating antibodies to Sarcocystis neurona in raccoons (Procyon lotor) from an urban area of Virginia. AB - Equine protozoal myeloencephalitis is the most important protozoan disease of horses in North America and is usually caused by Sarcocystis neurona. Natural and experimentally induced cases of encephalitis caused by S. neurona have been reported in raccoons (Procyon lotor) and raccoons are an intermediate host for this parasite. A 3-yr-long serological survey was conducted to determine the prevalence of agglutinating antibodies to S. neurona in raccoons collected from Fairfax County, Virginia, a suburban-urban area outside Washington, D.C. Samples from 469 raccoons were examined, and agglutinating antibodies (> or = 1:50 dilution) were found in 433 (92.3%) of the raccoons. This study indicates that exposure to S. neurona is high in this metropolitan area. PMID- 15357092 TI - Parvicapsula minibicornis in anadromous sockeye (Oncorhynchus nerka) and coho (Oncorhynchus kisutch) salmon from tributaries of the Columbia River. AB - The myxosporean parasite Parvicapsula minibicornis is described from adult sockeye and coho salmon during spawning migrations in tributaries of the Columbia River in Canada and the United States. These observations extend the known distribution of this parasite from the Fraser River drainage basin. The parasite was identified in Columbia River salmonids using polymerase chain reaction (PCR) and by in situ hybridization, but unlike in Fraser River salmon, it was not observed in conventional histological preparations of the kidney. Prevalence of the parasite determined by PCR was higher in spawning sockeye from the Fraser River than in those from the Okanagan River. Our ability to explain the relatively low prevalence and absence of clinical P. minibicornis infections in Columbia River salmon is hampered by our poor understanding of the life cycle of this parasite. PMID- 15357093 TI - Effect of hydrogen peroxide and other protease inhibitors on Cryptosporidium parvum excystation and in vitro development. AB - This study was undertaken to observe the effects of hydrogen peroxide on Cryptosporidium parvum oocysts with respect to protease activity in comparison to known protease inhibitors. In assessing the possible mechanisms of action of hydrogen peroxide, treatment effectiveness was analyzed using 3 assays and the potential roles of proteases and cations were considered. Treatment of C. parvum oocysts with hydrogen peroxide inhibited protease activity up to 50% compared with untreated controls. Treatment of oocysts with chemicals that affect sulfhydryls, including N-ethylmaleimide and dithiolthreitol, inhibited protease activity by >90%. Treatment of oocysts with these chemicals, along with the protease inhibitors, phenylmethylsulfonyl fluoride (PMSF), ethylenediamine tetraacetic acid, and cystatin, inhibited protease activity as well as in vitro excystation and infection in a cell culture assay. Several mechanisms may result in the successful inhibition of infection and excystation by hydrogen peroxide treatment, including: oxidation of oocyst wall proteins or lipids, chelating of cations necessary for infection, or hydroxyl radical-induced DNA damage to sporozoites, or both. PMID- 15357094 TI - Helminth parasites of Chapalichthys encaustus (Pisces: Goodeidae), an endemic freshwater fish from Lake Chapala, Jalisco, Mexico. AB - A total of 6 helminth species were recorded during helminthological examination of 50 Chapalichthys encaustus from Lake Chapala, Jalisco, Mexico. Helminth species identified included: Salsuginus sp. (an undescribed monogenea); Posthodiplostomum minimum (metacercariae); Cyclustera ralli (metacestode); Polymorphus brevis (cystacanth); Contracaecum sp. (nematode larvae); and Rhabdochona lichtenfelsi (adult nematode). Of these, 2 (Salsuginus sp. and R. lichtenfelsi) are specialist species. The observed species richness, individual parasite abundance, and diversity were low. Data suggest that host specificity is an important factor contributing to observed community composition and richness. Host feeding habits and helminth species availability seem to determine the characteristics of these helminth assemblages. PMID- 15357095 TI - Cloning, characterization, and functional expression of Taenia solium calreticulin. AB - Calreticulin is an endoplasmic reticulum protein involved in the homeostasis of intracellular Ca++ and other physiological processes. A complementary DNA clone containing the complete coding sequence of Taenia solium calreticulin (TsCRT) was isolated and characterized. Recombinant TsCRT was expressed in bacteria as a 50 kDa protein that specifically bound calcium when tested in a radioassay. The deduced amino acid sequence has 47-50% identity with other reported calreticulins. Poor recognition of TsCRT by human and pig sera with confirmed cysticercosis discourages its use for diagnosis of the disease. However, further characterization and localization studies could provide insights into the role of TsCRT in T. solium physiology and host-parasite interactions. PMID- 15357096 TI - Microarray data demonstrate that Trypanosoma cruzi downregulates the expression of apoptotic genes in BALB/c fibroblasts. AB - Parasites have been shown to up- and downregulate host apoptosis, most likely facilitating their ability to successfully establish an infection in the host. It has been demonstrated that pathogens modulate well-established pathways, leading to cell death, including induction of the Fas-FasL system to promote apoptosis. In contrast, it has also been shown that in other instances a decrease in host cell apoptosis results after the upregulation of genes in the Bcl-2 family. The present study examined the ability of Trypanosoma cruzi to modulate expression of host cell genes of the TNFR1 apoptotic pathway. Using microarray technology, gene expression was compared between uninfected BALB/c fibroblasts and T. cruzi infected BALB/c fibroblasts. After comparing expression patterns between uninfected and T. cruzi-infected BALB/c fibroblasts, it was concluded that genetic expression of genes in the TNFR1 apoptotic pathway is downregulated in T. cruzi-infected cells, indicating that in BALB/c fibroblasts the parasite decreases the expression of genes, leading to host cell apoptosis. PMID- 15357097 TI - The prevalence of Toxoplasma gondii antibodies in wild and captive cetaceans from Japan. AB - The seroprevalence of Toxoplasma gondii was investigated in wild and captive cetaceans from Japan. Antibodies against T. gondii were examined by both latex agglutination test (LAT) and indirect hemagglutination test (IHAT) for 77 serum or plasma samples obtained from 59 individuals of 6 species, including 2 hybrids. Antibody titers greater than 1:64 in LAT and greater than 1:640 in IHAT, indicative of the presence of T. gondii, were found in 11.9% of 59 individuals. In 7 samples that showed a positive reaction by IHAT, T. gondii titers were examined for each immunoglobulin (Ig) fraction separated by sucrose gradient centrifugation. The antibody peaks in each fraction were divided into 3 types, thought to be a reaction of IgM (type 1), IgG (type 2), and IgM with IgG (type 3). Type 1 was found in serum from a bottle-nosed dolphin (Tursiops truncatus) and a killer whale (Orcinus orca) sampled soon after capture off the Japanese coast in 1988; it was concluded that infection in the wild had occurred less than 15 yr before the study was performed. The prevalence of putative IgM and IgG antibodies from a captive-bred T. truncatus suggested that T. gondii infection also occurred in the aquarium. PMID- 15357098 TI - The Hymenolepis diminuta-golden hamster (Mesocricetus auratus) model for the evaluation of gastrointestinal anticestode activity. AB - A novel laboratory anticestode assay was developed using Hymenolepis diminuta in the hamster. The commercial anticestode compounds, praziquantel, bunamidine, and niclosamide were active against patent infections of Hymenolepis diminuta in golden hamsters (Mesocricetus auratus) when given orally at 3.125, 100, and 200 mg/kg, respectively. The gastrointestinal nematode anthelmintics, cambendazole and mebendazole, were active at 50 mg/kg. Rafoxanide (fasciolicide) was active at 25 mg/kg, the lowest level tested. The coccidiostat, nicarbazin, was active at experimental levels (800 mg/kg and up). The anthelmintic-ectoparasiticide (endectocide), ivermectin, was inactive against the tapeworm at 0.5 mg/kg, as expected. PMID- 15357099 TI - Comparative susceptibility of brown trout and rainbow trout to Discocotyle sagittata (Monogenea). AB - The susceptibility of brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) to the monogenean Discocotyle sagittata in the United Kingdom was assessed by experimental infection of naive fish. One month postinfection with 100 oncomiracidia/host, brown trout harbored significantly lower burdens (27.7 worms/host +/- 4.13 SE) than rainbow trout (47.8 worms/host +/- 3.90; P = 0.002). This indicates that the consistently lower prevalence and intensity of D. sagittata recorded in naturally infected farmed fishes reflects differences in susceptibility to the parasite. The outcome may be related to the comparatively short-term association of this parasite with rainbow trout (introduced to Britain in the 1880s) compared with the established native host-parasite association. PMID- 15357100 TI - Seroprevalence of Toxoplasma gondii in swine from slaughterhouses in Lima, Peru, and Georgia, U.S.A. AB - Toxoplasma gondii is an important pathogen transmitted by food, with raw or undercooked meat as the main foodborne source of toxoplasmosis. In Peru, 2-4 million people have antibodies to T. gondii. It is believed that more than 60 million people in the United States are infected with T. gondii. In this study, the prevalence of T. gondii in pigs from Peru and the United States was determined by Western blot. The presence of IgG antibodies to T. gondii from serum samples was determined. Blood samples were collected from 137 pigs at a slaughterhouse in Lima, Peru, and 152 pigs at a slaughterhouse in Georgia. Of the serum samples collected from swine, 27.7% (n = 38) from Peru and 16.4% (n = 25) from the United States were positive for T. gondii. Swine represent a significant source of human infection with T. gondii in Peru and the United States. PMID- 15357101 TI - Disseminated toxoplasmosis in a captive ring-tailed lemur (Lemur catta). AB - A 3-yr-old secundiparous female ring-tailed lemur presented to the Auburn University Small Animal Clinic with signs of dyspnea, lethargy, and anorexia. The animal died before she could be examined, and a full necropsy was immediately performed. Provisional necropsy findings included moderate pneumonia and hepatopathy. Acute interstitial pneumonia and focal hepatocellular necrosis were confirmed histologically. Lung impression smears, histopathology, electron microscopy, immunohistochemistry, and tissue culture isolation resulted in a diagnosis of acute disseminated Toxoplasma gondii infection, which was confirmed by polymerase chain reaction. The isolate of T. gondii was avirulent for mice and was named AU Tgl and genetically is type II. The source of the infection remains unclear, but speculation suggests contaminated fruit or blackbirds (Passeriformes: Icteridae) acting as transport hosts for oocysts from nondomestic felids and feral cats on the property. PMID- 15357102 TI - Fatal toxoplasmosis in a bald eagle (Haliaeetus leucocephalus). AB - Toxoplasma gondii tachyzoites were identified in the myocardium of a bald eagle (Haliaeetus leucocephalus) that died of necrotizing myocarditis. The diagnosis was confirmed by immunohistochemical staining with T. gondii-specific polyclonal antibodies. This is a new host record for T. gondii. PMID- 15357103 TI - Molecular characterization of the Leishmania braziliensis L6 ribosomal protein. AB - By screening a Leishmania braziliensis complementary DNA library with a pool of sera from leishmaniasis patients, the gene coding for L6 ribosomal protein was isolated. The sequence, genomic organization, and transcription of this gene are described in this article. The sequence analysis of the L. braziliensis L6 gene shows a single open reading frame, which codes for a protein of 192 amino acids (aa) with a hypothetical molecular mass of 20.9 kDa. The protein exhibits significant sequence similarity to L6 ribosomal proteins from higher eukaryotes and yeast. Thus, the L. braziliensis L6 protein contains 4 functional motifs, which are located at equivalent positions in other L6 ribosomal proteins described previously. Interestingly, the L6 ribosomal protein from L. braziliensis contains a specific region of 14 aa and a tyrosine kinase motif, which is absent in human and C. elegans L6 protein. The locus coding the L. braziliensis L6 ribosomal protein is formed by 2 gene copies arranged in tandem and located in a chromosome of approximately 0.9. Mb. The genes are actively transcribed as 2 polyadenylated transcripts of approximately 1.15 and 0.85 kb, which differ in their steady-state level and stability. PMID- 15357104 TI - Portrait of human tapeworms. AB - The tapeworms of the genus Taenia that infect human beings are T. solium, T. saginata and T. saginata asiatica. Taenia solium and T. saginata exhibit unequivocal features that characterize them; in contrast, only recent DNA studies, morphological characteristics, and epidemiological and sanitary aspects indicate that T. saginata asiatica is a subspecies of T. saginata. These 3 tapeworms occur in humans in their adult stage, and the intermediate hosts are pigs for T. solium and T. saginata asiatica and cows for T. saginata. Their identification is crucial considering the migratory increase from Asia to the Western Hemisphere and the fact that these tapeworms coexist in the same environment in Asia; furthermore, it is estimated that movement in both directions across the United States-Mexico border exceeds 200 million persons per yr, and thus, opportunities for acquiring and transporting T. solium infections are multiplied. It is not easy to distinguish among these tapeworms; therefore, a comparative diagram of the 3 parasites is shown in this article, which will facilitate their identification. All morphological features, some of which allow for identification, are clear and can be easily distinguished among the 3 tapeworms. PMID- 15357105 TI - Criteria for species determination in the 'revolutum' group of Echinostoma. PMID- 15357106 TI - Asthma education--home-based intervention. AB - In order for patients with asthma to maintain an adequate level of self management of the disease they must be provided initial and ongoing education. Education is a major tool in order to achieve this goal. This review discusses some of the components of asthma education that must be considered. There are also opportunities for the respiratory therapist to play a significant role in asthma disease management in the home. Education is a key tool in the management of asthma. The respiratory care professional can play a significant role in a home-based program. PMID- 15357107 TI - The predictive and diagnostic values of procalcitonin and C-reactive protein for clinical outcome in febrile neutropenic patients. AB - BACKGROUND: Procalcitonin (PCT) represents a new marker of systemic inflammatory reactions to bacterial infections. The main aim in this study was to determine the diagnostic value of PCT in predicting the clinical severity of febril neutropenic attacks, compare it with that of C-reactive protein (CRP), and clarify its importance in culture-positive attacks. METHODS: Between February 2001 and April 2002, 36 patients who were neutropenic due to various hematologic disorders and febrile were entered into the study. Blood samples were obtained on the first day of fever for the measurement of serum PCT and CRP levels. RESULTS: In clinically severe neutropenic fever attacks, means of serum PCT and CRP levels were measured as 0.93+/-1.33 ng/mL and 67+/-24 mg/L, while they were 0.37+/-0.23 ng/mL and 32+/-19 mg/L in clinically mild ones (p = 0.033 and p < 0.001). On the other hand, no statistical significance was found between culture-positive and negative attacks when either serum PCT or CRP levels were taken into consideration (p = 0.133 and p = 0.141). The specificity and positive predictive value of the serum PCT test for severe febrile neutropenia was higher than that of the serum CRP test (0.80 vs. 0.57 and 0.50 vs. 0.39). However, sensitivity and negative predictive value for CRP were higher than the values for PCT (1.00 vs. 0.40 and 1.00 vs. 0.73). Diagnostic value and positive likelihood ratio of CRP for severe febrile neutropenia were higher than those of PCT (71 vs. 67 and 2.32 vs. 2.00). CONCLUSIONS: PCT and CRP are comparable with each other in prediction of the clinical severity of febrile neutropenic attacks. Furthermore, serum CRP levels correlate with the duration of fever. PMID- 15357108 TI - The value of plasma levels of tumor necrosis factor-alpha and interleukin-6 in predicting the severity and prognosis in patients with congestive heart failure. AB - BACKGROUND: High plasma levels of pro-inflammatory cytokines play an important role in the pathophysiology of congestive heart failure (CHF). Therefore, we conducted a case-control study to determine the correlations between plasma levels of cytokines, i.e., tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-6, and the severity and mortality in patients with CHF. METHODS: One-hundred and 18 cases (62+/-15 years old) were classified into 3 groups: group 1 comprised 44 control cases with normal coronary arteriogram and left ventriculography and without valvular disorders or cardiomyopathy; group 2 comprised of 37 cases with mild CHF in New York Heart Association (NYHA) functional class (FC) II; group 3 had 37 cases with moderate/severe CHF in NYHA FC III or IV. Pre-catheterization plasma levels of TNF-alpha and IL-6 along with clinical and hemodynamic variables and follow-up data of cardiac death were assessed. RESULTS: Patients of group 3 had smaller body mass index, lower systolic and diastolic blood pressures, faster heart rates, higher left ventricular end-diastolic pressure and lowered triglyceride levels than the patients of groups 1 and 2. The plasma levels of TNF-alpha and IL-6 increased significantly in patients of group 3 in comparison with patients of groups 1 and 2 (both p < 0.001). Over the following 1.5 years, 13 patients died. Univariate analysis identified the following variables to be associated with poor prognosis: NYHA FC (p < 0.001), plasma TNF-alpha (p = 0.013), plasma IL-6 (p < 0.001), systolic blood pressure (p = 0.001), heart rate (p = 0.045) and left ventricular end-diastolic pressure (p = 0.021). Multivariate Cox regression analysis identified the independent predictors of cardiac death as FC (p = 0.007) and plasma IL-6 (p = 0.021). CONCLUSIONS: Our findings indicate that the plasma levels of IL-6 and TNF-alpha and especially the former, is a useful marker to correlate the progression of severity and late cardiac death in patients with CHF. PMID- 15357109 TI - Transcranial magnetic stimulation in patients with transient ischemic attacks. AB - BACKGROUND: By definition, transient ischemic attacks (TIAs) do not leave a neurological deficit beyond 24 hours after onset. However, a subgroup of TIA patients is characterized by persistent perfusion defect on single photon emission computed tomogram or infarction on brain computerized tomogram and magnetic resonance imaging. Here, we applied transcranial magnetic stimulation (TMS) to study whether TIA could produce persistent subclinical dysfunction for more than 24 hours. METHODS: The study included 23 TIA patients who had the criteria of hand weakness as one of their clinical manifestations. TMS was done twice in each TIA patient. The first time was during the period of 24-48 hours after onset and the second 7 days after onset. We studied the cortical motor threshold, the latencies and the amplitudes of the motor evoked potentials, the central motor conduction time, and the cortical silent period at the intensity of 1.5 times motor threshold with maximal voluntary isometric contraction. The recording was at the first dorsal interosseous muscle. RESULTS: There was no significant difference between the whole group of TIA patients and normal control. However, in the subgroup of TIA patients who had hand weakness more than 1 hour, they had increased motor threshold and prolonged cortical silent period during the first test. Both improved 1 week after onset. On the contrary, in TIA patients who had hand weakness less than 1 hour, their data were all within normal limits during the first and the second studies. CONCLUSIONS: Our results indicate that the motor function of TMS study will recover to full if the motor symptoms subside within 1 hour in TIA patients. Subclinical motor deficits may persist in TIA patients who have motor symptoms more than 1 hour. PMID- 15357110 TI - Correlation between urinary tract pure stone composition and stone morphology on plain abdominal film. AB - BACKGROUND: Crystallographic composition of urinary tract stone varies in several chemical groups and determines the degree of fragmentation to extracorporeal shock wave lithotripsy (ESWL) which has been widely used for the treatment of renal and ureteral calculi. Visual prediction of stone composition by its morphology from plain radiograph may provide a simple method and clinical hint to decide therapeutic modalities. A prospective study was designed to determine the correlation of stone composition with its radiographic morphology. METHODS: One hundred patients of urolithiasis with passage of stone fragments (after ESWL, surgical lithotomy or endoscopic lithotripsy) analyzed as pure stone by infrared spectrometry were reviewed by plain abdominal film to classify their morphology, location and size. Five experienced surgeons familiar with the radiographic patterns of different types of pure stones were invited to predict the stone composition on plain abdominal film. RESULTS: Five different patterns of radiographic appearances of the stones were identified. Calcium oxalate monohydrate referred to the smooth-shaped and homogeneous ones with denser opacity. Calcium oxalate dihydrate referred to those smaller ones that had irregular margin with stippled border and were less dense. Calcium phosphate (CaP) had moderate density and size without significant figures. Struvite meant staghorn stone; usually bilateral. Uric acid referred to those being radiolucent, slightly opacified, and in large size. The 5 surgeons achieved an overall accuracy of 27.4%. There was no obvious relations between accuracy and location or size of the stones. CaP stone was the most likely to be misclassified, and achieved only 20% accuracy. CONCLUSIONS: Different stone composition showed grossly specific radiographic appearances, but clinical test revealed poor correct rate in prediction of stone composition from plain abdominal film. The results of our study suggest that prediction of stone component based on stone morphology on plain abdominal film may not be accurate enough. Patient's clinical information and other laboratory data should be considered while interpreting plain abdominal film for better treatment choice of urolithiasis. PMID- 15357111 TI - The relationship between parental concerns and professional assessment in developmental delay in infants and children--a hospital-based study. AB - BACKGROUND: Developmental delay (DD) in infants and children is one of the chief complaints of parents. It has been established that the concerns of parents are as accurate as quality screening tests. Some kinds of concerns are particularly useful in the early detection of associated developmental problems. The purpose of this study was to investigate the relationship between the main parental concerns and the final diagnosis based on professional assessment of children who are suspected to have DD. METHODS: One-hundred and 1 infants or children were recruited into this study. The major concerns of parents were elicited and categorized by various developmental domains: speech, motor, behavioral, cognitive, global, and non-specific developmental problems. All children underwent comprehensive combined assessments by professionals in the hospital, and were classified into 6 subtypes: speech, motor, behavioral, cognitive, and global DD, and normal development. RESULTS: Our results revealed that parental concerns about speech, motor, and behavioral development yielded a high sensitivity to the final diagnosis of the same developmental domain (77-89%). However, concerns about cognitive and global DD had limited sensitivity (15-36%). On the other hand, concerns about global, speech, and motor DD had relatively higher positive predictive values (55-77%). Comparatively, cognitive or behavioral concerns had lower positive predictive values (25-33%). CONCLUSIONS: Such results indicate that parents play an important role in detecting speech, motor and behavioral DD in children. Parental concerns about cognition and behavior should be questioned in terms of their association with the real problem. This information may be useful in establishing trends in understanding the discrepancy or relationship between parental concerns and professional assessments in DD. PMID- 15357112 TI - Fenofibrate-induced acute cholestatic hepatitis. AB - Fenofibrate-induced acute or chronic hepatitis is rare, and only 11 reports from French, Italian or Spanish literature have been published up to date. We report a case of fenofibrate-induced acute cholestatic hepatitis. To the best of our knowledge, this is the first one reported in Taiwan. A 61-year-old man developed acute cholestatic hepatitis after taking fenofibrate 100 mg tid for 10 days. Laboratory profile on admission showed serum total bilirubin 9.3 mg/dL, direct bilirubin 2.7 mg/dL, alanine aminotransferase 249 IU/L, aspartate aminotransferase 259 IU/L, alkaline phosphatase 259 IU/L, and gamma-glutamyl transpeptidase 1014 IU/L. Pathology proved hepatocanalicular cholestasis in liver. Fenofibrate was discontinued immediately. His clinical manifestations and liver function tests improved gradually and returned to nearly normal in 2 months. We suggest that liver function tests, including total bilirubin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and gamma glutamyl transpeptidase, be checked at least 2 weeks after taking the drug and that serum aminotransferase be monitored every 3 months during the first 12 months of therapy. Treatment should be discontinued if alanine aminotransferase values increase by more than 100 IU/L. PMID- 15357113 TI - Prolonged survival after single lung transplantation for pulmonary hypertension secondary to systemic lupus erythematosus. AB - Pulmonary hypertension is one of the life-threatening complications of systemic lupus erythematosus, but these patients are often excluded from lung transplantation candidacy due to the nature of underlying multiple system involvement. We report a long-term survival after single lung transplantation in a case of systemic lupus erythematosus with severe pulmonary hypertension. It suggests that single lung transplantation may be considered in such patients, especially in condition of limited donor organ supply. PMID- 15357114 TI - Surgical eradication of esophageal carcinoma in a patient with Klippel-Trenaunay syndrome. AB - Klippel-Trenaunay syndrome (KTS) is a rare congenital vascular disease of unknown etiology, and its occurrence with malignant tumor is extremely rare. We herein report a case of KTS presenting chronic gastrointestinal bleeding and complicating with adenosquamous cell carcinoma (ASCC) of the esophagus. The therapeutic dilemma of managing ASCC of the esophagus in a patient who combined with the rare congenital syndrome posed a difficult and interesting clinical problem. However, ASCC is also a very rare histological tumor in the esophagus, and its relationship to KTS has never been reported. In this article, we reviewed all the reported malignancies related to KTS. The possible surgical risks and complications, as well as preoperative imaging investigation, are also discussed. PMID- 15357115 TI - Oncocytic Schneiderian papilloma. AB - Oncocytic Schneiderian papilloma, fungiform papilloma, and inverted papilloma are 3 morphologically separate tumors arising from the Schneiderian membrane. Oncocytic Schneiderian papillomas comprise about 3 approximately 5% of this entity. Old-aged group predominates. No sex predilection is noted. Multi-layered eosinophilic epithelium characterizes this kind of tumor. Exophytic and inverted growth pattern is found microscopically. It is often confused with sinonasal adenocarcinomas which contain only single-layered epithelium. Clinically, its behavior parallels inverted papillomas due to local recurrence and coexistence of malignancy. We reported a case of oncocytic Schneiderian papilloma arising from the anterior ethmoid sinus and extending to maxillary sinus antrum. Although destruction of the lamina papyracea was noted preoperatively, no malignancy was found microscopically. We adopted endoscopic approach and removed the tumor thoroughly. External approach was necessary if tumor extent was not feasible to endoscopic approach. The etiology of oncocytic Schneiderian papillomas remains unknown. However, it should be considered in the differential diagnosis of unilateral nasal polypoid lesions clinically. PMID- 15357116 TI - Idiopathic spinal epidural lipomatosis. AB - Spinal epidural lipomatosis is a rare disorder characterized by the pathological overgrowth of epidural fat and often causes extradural compression which may mimic other spinal disorders. Steroid administration is a well-documented etiology. We report a case of spinal epidural lipomastosis without history of steroid administration. The initial manifestation was bizarre and was misdiagnosed as a degenerative vertebral or disc disease for long. Posterior decompression with debulking of the extradural fat successfully and thoroughly relieved his symptoms. We emphasize the importance of taking this disease into consideration for a markedly obese patient with persisting back pain or symptoms suggesting spinal cord or other spinal neural elements compression. PMID- 15357117 TI - As the patient voice grows louder, it is time to see who's listening. PMID- 15357118 TI - Chronic disease ideas. Too tight a spotlight? AB - Greater emphasis on chronic-disease management is welcome, but focusing too much on US models may mean overlooking valuable lessons from the UK, including those on diabetes. PMID- 15357119 TI - Clinical management where medicine meets management. Down to earth. AB - Appreciative inquiry is a method that can help NHS staff during periods of change management. Staff turnover and sickness absence are already down at one heart centre that has used the technique. People who have attended the away-days say approachability to senior staff is improved. PMID- 15357120 TI - Delayed discharge. Must get out more often. AB - Delayed discharge fines increase pressure on trusts to prevent delays. The loss of vital information between health and social services undermines discharges' credibility. Pooled budgets between the two sectors are a powerful means of fostering innovation. PMID- 15357121 TI - Law. Isolated cases. PMID- 15357123 TI - My brilliant career--finance director. In-depth analysis. Interview by Seamus Ward. PMID- 15357124 TI - 'Theatre of the future' arrives. PMID- 15357125 TI - Questioning oxygen flow rate guidance. PMID- 15357126 TI - Collaborative effort triumphs in project. PMID- 15357127 TI - Full coordination is key to project success. AB - The procurement of complex and highly serviced hospital buildings is always fraught with the expensive and complicated problem of inadequate coordination of building services. Obviously, this problem is detrimental to the success of any project. All project practitioners must respond positively to increasingly complex building design and construction and higher expectations of their clients. To give greater value improvement in construction, the present way of managing building services design and installation must be "re-engineered" whether we use traditional or nontraditional procurement methods. This will require a cultural shift in our industry as a whole, away from adversarial relationships to more cooperative and partnering relationships. The author's research based on case studies in Hong Kong indicates that there is no best procurement practice. Rather, there is appropriate management of building design and construction. The results also show that good building services coordination can only be achieved by a good building team having a fully integrated and coordinated design, an appropriate integrating mechanism, a fair contract, effective management of coordination, mutual trust and respect, highly integrated supply chain management, greater co-operation and involvement, efficient managerial control of the whole design and construction process and the use of re engineering of construction. All these attributes can give significant improvement in time, cost and quality performance needed by each member of the building team. PMID- 15357128 TI - Cutting carbon emissions. PMID- 15357130 TI - Fast route to creating children's centres. New, specialised centres for children must have challenging. PMID- 15357129 TI - Comparing protective measures in Europe. PMID- 15357131 TI - Improving patient access to the Veterans Health Administration's primary care and specialty clinics. AB - BACKGROUND: During the past four years the Veterans Health Administration (VHA) has been engaged in a national effort to improve access for patients to its 1,826 primary care, audiology, cardiology, eye care, orthopedics, and urology clinics by using the principles of open access or "advanced clinic access." The strategy entailed the development of successful cases to demonstrate the methods of advanced clinic access and provide evidence of its benefits for providers as well as patients in primary care and specialty clinics. RESULTS: Four clinics--one primary care clinic and three specialty care clinics--showed dramatic improvement in waiting times for appointments (reductions range from 20 days in urology to 78 days in primary care). DISCUSSION: Beyond the four case studies, hundreds of other clinics in the VHA are also applying advanced clinic access principles in their work. The diversity across the VHA suggests that the principles of advanced clinic access are robust across settings and types of clinics. However, the experience of other organizations with different structures and patient populations needs to be reported to fully demonstrate the generalizability of these results. Many of the changes were put in place during the project's final 18 months. Additional data will be needed to demonstrate sustained improvement. PMID- 15357132 TI - Implementing a hospitalwide patient safety program for cultural change. AB - BACKGROUND: After focus groups revealed that staff perceived a punitive culture, Missouri Baptist Medical Center (MBMC) embarked on a comprehensive patient safety program, which was initially directed at creating a just culture of patient safety. INTERVENTIONS: A series of structures, processes, and initiatives were introduced to change the attitudes of management and staff toward human error, to communicate broadly with staff and the community, and to provide feedback on leadership's responses to specific events. All events reported were tracked continuously and recorded each month on a spreadsheet. RESULTS: Total medical events reported by staff increased significantly (p < .001) from 35 to 132 per 1,000 patient days. Reports to the hotline alone increased significantly (p < .001) from 3 to 23 per 1,000 patient days, and the proportion of callers who left their name increased significantly (p < .001) from 30% to 61%. Survey results from staff showed a small but significant increase in awareness of patient safety and in comfort with reporting. CONCLUSION: The implementation of a carefully planned and orchestrated series of interventions designed to improve a hospital's culture of patient safety can, if led by senior hospital executives, lead to a substantial, profound, and lasting increase in error reporting and improvement in employee perceptions of the organization's safety culture. PMID- 15357133 TI - Strategies for increasing adherence to clinical guidelines and improving patient outcomes in small primary care practices. AB - BACKGROUND: The best way to get research findings into practice needs to be determined, particularly in small practices. The Practice Partner Research Network (PPRNet) is a nationwide practice-based research network of small primary care practices that use the same electronic medical record (EMR). Between 2000 2003 the PPRNet Translating Research into Practice (TRIP) project tested a multimethod intervention to help practices improve primary and secondary prevention of cardiovascular disease and stroke. Intervention sites each hosted six to seven site visits and participated in two annual network meetings during the two-year intervention period. A model describing practice-based improvement strategies was validated using prospective data from 10 intervention and 9 control sites. RESULTS: The model consisted of five categories of improvement strategies: Prioritize Performance, Involve All Staff, Redesign Delivery Systems, Activate Patients, and Use EMR Tools. PPRNet-TRIP intervention practices used more of the model items than did controls (69% versus 48%, p = .053), as did high performing practices versus mid-range or low performers (81% versus 39% versus 46%, p = .001). CONCLUSION: The PPRNet-TRIP Improvement Model might guide small practices in their efforts to translate research into practice and improve care outcomes. PMID- 15357134 TI - Remote order entry and video verification: reducing after-hours medication errors in a rural hospital. AB - BACKGROUND: Hendry Regional Medical Center, a 66-bed facility, has 8.5 hours of pharmacy staffing daily. Review of two years of medication errors indicated that 34% of all medication errors involved selection of the wrong drug from stock after pharmacy hours. PROCESS CHANGES: During 2003 many changes were implemented in the medication management process to enhance medication safety. For example, a limited-access area of the pharmacy was established for night entry by the nursing supervisor. The hospital then implemented services with a newly licensed remote order-entry service. These steps decreased the rate of errors involving wrong drug selection to 15%. Subsequently, a medication error involving selection of the wrong drug at night prompted another look at the process. IMPLEMENTING THE VIDEO VERIFICATION PROCESS: The hospital began using a webcam to create a digital image of the medication selected for removal from the pharmacy stock by the nursing supervisor. This image is transmitted via e-mail. In six months, only one (recent) error resulting from selection of the wrong medication after normal pharmacy hours occurred; the medication selection process simply was not followed. DISCUSSION: Results of medication error reporting indicated that using Webcam/e-mail image transmission and video verification by a pharmacist further improved patient safety in the medication selection process. PMID- 15357135 TI - Using self-report to improve substance abuse risk assessment in behavioral health care. AB - BACKGROUND: Primary care and behavioral health clinicians frequently fail to detect substance-related problems among their patients, thereby leading to increased morbidity and mortality and health care costs. A managed behavioral health care organization(MBHO) conducted a quality improvement initiative in which clients seeking outpatient psychotherapy were screened by self-report for substance-related problems, and clinicians were provided with feedback in cases of discrepant findings. METHOD: Client self-report questionnaires, which included items inquiring regarding problems related to substance abuse, were administered at multiple points during treatment episodes. Clinicians were also asked to complete assessments, including indicating the presence of a substance abuse problem. RESULTS: Clinicians failed to identify substance abuse problems in > 80% of the cases where the patient endorsed items clearly related to substance abuse on the outcome questionnaire. In the quality improvement intervention, the MBHO sent letters alerting clinicians to the clients' self-reported substance abuse problems. The concordance between clinician assessment and client self-reported problems then increased significantly. DISCUSSION: Results of the study argue for the utility of using client self-report measures as part of a comprehensive effort to measure and improve the effectiveness of behavioral health care treatment services. PMID- 15357136 TI - An overlooked alliance: using human factors engineering to reduce patient harm. AB - BACKGROUND: Although human factors engineering (HFE) is considered only in relationship to the design of medical devices or information systems technology, human factors issues arise in many aspects of work in health care organizations. HFE ANALYSIS: In one scenario, the resuscitation stretcher would not pass through the ED door closest to radiology. Many clinical work spaces were never formally designed for the work currently being performed in them; instead, they were adapted from existing space originally designed for a different use. In a second scenario, infusion pump malfunction was not apparent. The patient experienced a near miss secondary to poor design; users thought that the infusion pump had been turned off when it was not. RECOMMENDATIONS: Health care can significantly benefit from the incorporation of HFE into the workplace. Introductory classes in medical and nursing schools on HFE will assist students in detecting HFE-related issues, making them less likely to suffer with them or overlook them once in clinical practice. More extensive training for patient safety and risk managers, that is, at a minimum, a certificate-level course from an HFE program, would enhance case and root cause analyses since these issues are rarely factored in. CONCLUSION: Collaboration with HFE experts and use of HFE principles may not make health care fool-proof, but it will make it less dependent on improvisation and ingenuity to protect patients from the system's vulnerabilities. PMID- 15357137 TI - Closing the communication loop: using readback/hearback to support patient safety. PMID- 15357138 TI - Promoting a healing environment: quiet time in the intensive care unit. AB - A midafternoon "quiet time" period was instituted to promote patients' recovery and healing. PMID- 15357139 TI - Protective role of metallothioneins in the injured mammalian brain. AB - Metallothioneins (MTs) are small cysteine-rich proteins which are found widely throughout the mammalian body, including the CNS. There are extensive data on the structure and expression of MTs, and many basic properties pertinent to MT biology in the CNS appear to be well established. As discussed in this review, one isoform class (MT-I/II) is rapidly induced following many types of CNS insult, and is strongly neuroprotective, whilst another isoform class (MT-III) shows major differences in its expression profile and physiological properties. As in other tissues, there is no clear consensus on the mechanism of MT action in the CNS and how it exerts its protective role, despite a number of excellent animal and cell culture models of MT expression in the brain, and a large literature on the physico-chemical properties of MTs extending over several decades. This review is therefore an attempt to summarise the recent literature on the expression of MTs in the adult mammalian brain and how MTs possibly act to protect the brain following physical or chemical insult. One exciting finding from recent work is that perturbing the levels of MT in the brain has an effect that extends beyond cells which normally express MT to other cell types including neurons, microglia and cells of the immune system. These observations were made mainly using animal models in which MT action can be observed in its normal cellular context, and this review focuses particularly on work conducted in animal models of physical and chemical injury in the brain. PMID- 15357140 TI - Shining light on neurons--elucidation of neuronal functions by photostimulation. AB - Many neuronal functions can be elucidated by techniques that allow for a precise stimulation of defined regions of a neuron and its afferents. Photolytic release of neurotransmitters from 'caged' derivates in the vicinity of visualized neurons in living brain slices meets this request. This technique allows the study of the subcellular distribution and properties of functional native neurotransmitter receptors. These are prerequisites for a detailed analysis of the expression and spatial specificity of synaptic plasticity. Photostimulation can further be used to fast map the synaptic connectivity between nearby and, more importantly, distant cells in a neuronal network. Here we give a personal review of some of the technical aspects of photostimulation and recent findings, which illustrate the advantages of this technique. PMID- 15357141 TI - Expression and function of brain specific small RNAs. AB - Small non-messenger RNAs (snmRNAs) are a heterogeneous group of non-coding RNAs with a variety of regulatory functions including regulation of protein expression and guidance in RNA modifications. They are actively being investigated in Archaebacteria, yeast, invertebrates and mammals. Brain-specific snmRNAs have been identified in mammals and they seem to contribute to neuronal differentiation during development and to brain functions subserving learning and memory. Here we review the current knowledge of the properties, expression and functions of three groups of brain-specific snmRNAs: small nucleolar RNAs, BC1/BC200 RNAs and microRNAs. PMID- 15357142 TI - Direct and indirect effects of corticosteroids on astrocyte function. AB - Corticosteroids are used for a variety of conditions; among the most well-known uses are for asthma and eczema. We review here the direct and indirect effects of corticosteroids on astrocyte physiology. Astrocytes play an important role in communication between neural cells, as one astrocyte can communicate with many neurons. They are also central in bringing nutrients through the blood-brain barrier (BBB) to the brain areas they serve. Therefore, any chemical or pharmaceutical product entering the brain via the BBB will first come into contact with the astrocytes. We discuss the direct effects that corticosteroids have on astrocyte physiology and functioning; these include inhibited glucose transport, decreased glycogen synthesis and decreased glutamate uptake. Furthermore, the indirect effects of corticosteroids on astrocytes are also reviewed. We know that corticosteroids lower neural serotonin. Lowered serotonin affects astrocyte functioning, and particularly astrocytic cAMP activities, a decrease in cytokine activities and impaired GABA uptake. These can be seen as the indirect effects of corticosteroids on astrocyte physiology. Corticosteroids therefore have a pertinent effect on neuro-energetics due to astrocyte physiology impairment, and this may ultimately be the reason for memory impairment of patients who chronically use corticosteroids. PMID- 15357143 TI - Genetic aspects of melatonin biology. AB - For decades, the important physiological roles of the pineal hormone have inspired scientific investigations. Research efforts have generated a broad amount of information relevant to various genetic aspects of melatonin biology. Nevertheless, our understanding of the effect of genetic factors upon melatonin biosynthesis and the mechanisms of gene expression regulation by melatonin in target tissues is far from complete. The present review makes an effort to summarize and systematize the existing information on the subject, sequentially discussing (i) the effect of genetic factors upon melatonin biosynthesis, (ii) melatonin receptor expression profiles, and (iii) the effect of melatonin upon expression of genes in target tissues. PMID- 15357144 TI - A new conceptual model for assistive technology outcomes research and practice. AB - Predictive conceptual models help us frame research questions, interpret results, and guide clinical practice. Although numerous models have appeared in the assistive technology (AT) literature, none has been shown to predict AT usage. The lack of a valid predictive model indicates the need for development of new approaches to modeling AT outcomes. This article proposes a user-centered conceptual model that predicts AT usage as a function of the perceived relative advantages of AT. Device usage is not modeled as a one-time, all-or-nothing proposition, but as a decision process recurring over time. The influence of parallel interventions working concurrently with, or as an alternative to, AT is a central consideration that ultimately drives AT usage. Usage is shown as a proximal influence on AT impact, and AT impact is shown to be a predictor of future use. Research is cited supporting various elements of the new model. PMID- 15357145 TI - Augmentative and alternative communication and cell phone use: one off-the-shelf solution and some policy considerations. AB - Use of cell phones has grown dramatically; however, there remains a gap between people with disabilities and the general public in regard to access to cellular telephony. This article describes the importance of cell phone use among people who use augmentative communication devices. An "off-the-shelf" solution is described and illustrated with the goal of improving access to mobile communication among individuals with significant communication disabilities. Finally, a discussion of the need for universal design of cell phones that includes such features as infrared ports, volume range, speakerphones, matrix displays, EZ buttons, voice dialing, and messaging is presented. PMID- 15357146 TI - Outcomes of wheelchair systems intervention with residents of long-term care facilities. AB - This pilot study was designed to measure the effects of individually prescribed wheelchair systems on posture and reach, mobility, quality of life, and satisfaction with technology for residents of long-term care facilities. Thirty persons 60 years of age or older who resided permanently in a long-term care facility and who used seating and mobility systems for 6 hours or more each day were recruited for this project. Outcomes included timed independent mobility, forward and lateral reach, quality of life, and satisfaction with assistive technology. The study used semicrossover design with participants measured three times. Measurements were first made in the existing seating and mobility system and a second time immediately after participants were provided with individually prescribed seating and mobility systems. The final measurement was 3 months after the delivery of the individually prescribed system. Results indicated that individually fitted wheelchair systems for elderly residents of long-term care facilities are beneficial. Participants had less difficulty independently propelling their systems and increased forward reach, quality of life for social function and physical role, and satisfaction with the new wheelchair technology. Persons residing in extended care facilities benefit from receiving individually prescribed wheelchair systems. The individual systems enhance elderly persons' independent mobility, functional reach, feeling of well-being, and satisfaction with their assistive technology. PMID- 15357147 TI - Using telerehabilitation to identify home modification needs. AB - Teleconferencing technology has great potential for providing cost-effective in home assessment for home modification services from anywhere to anyone in need. Despite its enormous potential, the use of this technology as a means to deliver these specific services had not been investigated. This project investigated the use of televideo technology to provide remote home assessment services to patients prior to discharge so that they could function as independently as possible in their own homes after being discharged from a specialty clinic. Specifically, an assessment protocol that could be implemented using video conferencing technology was developed and feasibility of the remote assessment process was determined by validating it against the standard of practice, an in home assessment by a home modifications specialist. Independent in-home and remote home assessments were completed by two occupational therapists who specialize in home modifications. The results were compared for agreement in identification of specific accessibility problems in and quantitative measurements of the home. The remote assessment correctly identified a total of 51 of the 59 problems (86.4%) identified by the in-home assessment and only identified five problems (8.9%) that were not identified by the on-site assessment. In addition, 54 of 60 (90%) of the quantitative measurements from the remote assessment matched those from the in-home assessment. Findings suggest that remote telerehabilitation assessments have the potential to enable specialists to diagnose potential accessibility problems in home environments and prescribe appropriate modifications regardless of the location of the client, home, or specialist. PMID- 15357148 TI - Using immersive technology for postural research and rehabilitation. AB - Posture has traditionally been examined by isolating individual control pathways to determine their specific contributions. However, if these pathways are responsive to functional contexts, then their responses may differ when the system is receiving simultaneous inputs from multiple pathways. Thus, we may never fully understand how the central nervous system (CNS) organizes behaviors in the real world from studies conducted in the minimized environment of the laboratory. The consequence of this is that when findings from the laboratory are applied to therapeutic intervention, the intervention may not be appropriate for all circumstances and will not fully meet the needs of the patient. We have united an immersive dynamic virtual environment with motion of a posture platform to record the biomechanical and physiological responses to combined visual, vestibular, and proprioceptive inputs. The virtual environment possesses content, contrast, and texture so that we can examine postural responses as they might occur in a complex, real-world environment. In this paper we specifically describe the factors guiding our choices of virtual technology and present data from young adults, elderly adults, and an individual with bilateral labyrinthine loss to demonstrate how multimodal inputs influence their postural response organization. Significant implications for future experimental and rehabilitation protocols are also discussed. PMID- 15357149 TI - Users' perceptions of the impact of electronic aids to daily living throughout the acquisition process. AB - This study investigated the experience of seven new users of a particular type of assistive technology through the stages of anticipating, acquiring, and using an electronic aid to daily living. A mixed methods research approach was used to explore each of these stages. The Psychosocial Impact of Assistive Devices Scale was used to measure the perceived impact of the new assistive technology on users' quality of life, and findings were further explored and developed through open-ended questioning of the participants. Results indicated that preacquisition of the device, users predicted that the electronic aid to daily living would have a positive impact on their feelings of competence and confidence and that the device would enable them in a positive way. One month after acquiring the device a reduced, yet still positive, impact was observed. By 3 and 6 months after acquisition, perceived impact returned to the same positive high level as preacquisition. It is suggested that prior to receiving the device, potential users have positive expectations for the device that are not based in experience. At the early acquisition time, users adjust expectations of the role of the assistive technology in their lives and strive to balance expectations with reality. Three to 6 months after acquiring an electronic aid to daily living, the participants have a high positive view of how the device impacts on their lives based in experience and reality. A model illustrating the electronic aids to daily living acquisition process is proposed, and suggestions for future study are provided. PMID- 15357150 TI - Co-infection or co-colonization with vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus in a network of community hospitals. PMID- 15357151 TI - Nosocomial candidemia: an ounce of prevention is better than a pound of cure. PMID- 15357152 TI - Has the epidemiology of nosocomial candidemia changed? AB - OBJECTIVE: To assess changes in the epidemiology of nosocomial candidemia in the post-fluconazole era among hospitalized patients using a case-control study design. DESIGN: Candidemia case-patients were matched 1:1 on diagnosis, age, and length of stay with control-patients. Conditional logistic regression was used to determine predictors and outcomes of candidemia. Treatment regimens and compliance with national practice guidelines were compared among case-patients. SETTING: Barnes-Jewish Hospital, a 1,278-bed, tertiary-care center affiliated with Washington University School of Medicine, St. Louis, Missouri. PARTICIPANTS: Patients admitted from January 1 to December 31, 2000. Case-patients were identified through the hospital microbiological surveillance system and matched with control-patients. RESULTS: Predictors of candidemia included Hickman catheters (odds ratio [OR], 9.53; 95% confidence interval [CI95], 1.34 to 68.01), gastric acid suppressants (OR, 6.38; CI95, 2.33 to 17.43), nasogastric tubes (OR, 3.69; CI95, 1.27 to 10.78), antibiotics (OR, 1.46; CI95, 1.15 to 1.86), and admission to the intensive care unit (OR, 6.40; CI95, 2.12 to 19.31). The crude case-fatality rate was 40%. Seventeen (15%) of the case-patients received the recommended treatment regimen according to recently published practice guidelines. CONCLUSIONS: The epidemiology of candidemia has changed little at our hospital during the past decade and remains a significant cause of mortality. Further studies on the benefits of preventive therapy will be essential to improve the outcome of this infection. PMID- 15357153 TI - Genetic relatedness among Candida tropicalis isolates from sporadic cases of fungemia in two university hospitals in Korea. AB - OBJECTIVE: To compare the epidemiology and genetic relatedness of Candida tropicalis isolates causing bloodstream infection (BSI) in two hospitals. SETTING: Two tertiary-care hospitals in Korea. METHODS: A retrospective molecular epidemiologic analysis using pulsed-field gel electrophoresis (PFGE) was performed with 49 C. tropicalis isolates from sporadic cases of BSI. The isolates were collected from 27 patients at Chonnam National University Hospital (CUH) during a 6-year period and 22 patients at Asan Medical Center (AMC) during a 2 year period. RESULTS: Based on the PFGE patterns, the average similarity value (S AB) for the 27 isolates from CUH was 0.84 +/- 0.08, which was significantly higher than that for the 22 isolates from AMC (0.78 +/- 0.06; P < .001). Of the 49 strains from patients at the 2 hospitals, 9 isolates were placed into 3 subtypes with S AB values of 1.0, which indicated that they were identical. All 9 of these strains were isolated from CUH patients, and each type strain was isolated sporadically during a period ranging from 4 months to 3 years. On comparison of the clinical characteristics of the patients of the 2 hospitals, the CUH strains were isolated more frequently from non-neutropenic patients and patients with central venous catheter-related fungemia; cases from CUH had a better outcome than those from AMC (P < .05). CONCLUSIONS: These data show that the clinical and epidemiologic characteristics of C. tropicalis fungemia may differ markedly among hospitals and that some cases of C. tropicalis fungemia may be caused by endemic strains within a hospital. PMID- 15357154 TI - Candida parapsilosis bloodstream infection in pediatric oncology patients: results of an epidemiologic investigation. AB - OBJECTIVE: To investigate an outbreak of Candida parapsilosis bloodstream infections (BSIs) involving three patients admitted to a pediatric oncology unit between April and June 2002. METHODS: After the third case was documented, cultures were performed of the hands of all medical and paramedical staff members in the pediatric oncology unit and of environmental surfaces in the rooms occupied by the three patients. Electrophoretic karyotyping with pulsed-field gel electrophoresis and arbitrarily primed polymerase chain reaction were used to assess the genetic relatedness among C. parapsilosis isolates. RESULTS: The three cases of C. parapsilosis BSI were diagnosed based on blood cultures performed during a 38-day period. Evidence of prior C parapsilosis colonization of the gastrointestinal tract was present in only the first case. Each patient had an indwelling central venous catheter (CVC), which was promptly removed, and semiquantitative catheter tip cultures also revealed C. parapsilosis. None of the 30 environmental cultures performed was positive for C. parapsilosis, but the fungus was isolated from the hands of 6 of the 20 nurses tested. Both molecular typing methods revealed identical DNA fingerprinting patterns for all 13 patient isolates (7 from blood, 3 from CVC tips, and 3 from the gastrointestinal tract) and for 5 of the 6 recovered from the nurses' hands. CONCLUSIONS: These findings suggest the possibility of cross-infection with a single C. parapsilosis strain that was transmitted (probably during CVC dressing changes) by nurses whose hands were colonized with it. The role of previous gastrointestinal colonization in the first case cannot be excluded. PMID- 15357155 TI - Central venous catheter-related bacteremia due to gram-negative bacilli: significance of catheter removal in preventing relapse. AB - OBJECTIVE: To study the characteristics of catheter-related, gram-negative bacteremia (GNB) and the role of central venous catheter (CVC) removal. DESIGN: This retrospective study involved a search of the microbiological department records of CVC and blood cultures and patients' medical records. SETTING: University of Texas M. D. Anderson Cancer Center, a tertiary-care hospital in Houston, Texas. PATIENTS: Patients with cancer who had catheter-related GNB, defined as (1) a positive catheter tip culture with at least 15 colony-forming units semiquantitatively, (2) isolation of the same organism from the tip and peripheral blood cultures, (3) no other source for bacteremia except the CVC, and (4) clinical manifestations of infection (fever or chills). RESULTS: Between January 1990 and December 1996, 72 cases of catheter-related GNB were available for review. Most of the patients (67; 93%) had their CVCs removed in response to the bacteremia. Few patients (5; 7%) retained their CVCs and were treated with appropriate antibiotics. When CVCs were removed, only 1 patient (1%) relapsed with the same organism, whereas all 5 patients with retained CVCs relapsed after having responded (P < .001). The most commonly isolated organisms were Enterobacter, Klebsiella, Stenotrophomonas, Pseudomonas, and Acinetobacter species. Catheter removal within 72 hours of the onset of the catheter-related GNB was the only independent protective factor against relapse of the infection (odds ratio, 0.13; 95% confidence interval, 0.02-0.75; P = .02). CONCLUSION: In patients with documented catheter-related GNB, CVCs should be removed within 48 to 72 hours to prevent relapse. PMID- 15357156 TI - Routine changing of intravenous administration sets does not reduce colonization or infection in central venous catheters. AB - OBJECTIVE: To determine the effect of routine intravenous (IV) administration set changes on central venous catheter (CVC) colonization and catheter-related bacteremia. DESIGN: Prospective, randomized, controlled trial. SETTING: Eighteen bed intensive care unit (ICU) in a large metropolitan hospital. PARTICIPANTS: Two hundred fifty-one patients with 404 chlorhexidine gluconate and silver sulfadiazine-coated multi-lumen CVCs. INTERVENTIONS: CVCs inserted in the ICU and in situ on day 4 were randomized to have their IV administration sets changed on day 4 (n = 203) or not at all (n = 201). Use of fluid containers and blood product administration sets was limited to 24 hours. CVCs were removed when not required, infection was suspected, or in place on day 7. Catheter cultures were performed on removal by blinded laboratory staff. Catheter-related bacteremia was diagnosed by a blinded intensivist using strict definitions. Data were collected regarding catheter duration, site, Acute Physiology and Chronic Health Evaluation (APACHE) II score, patient age, diagnosis, hyperglycemia, hypoalbuminemia, immune status, number of fluid containers and IV injections, and administration of propofol, blood, total parenteral nutrition, or lipid infusion. RESULTS: There were 10 colonized CVCs in the group receiving a set change and 19 in the group not receiving one. This difference was not statistically significant on Kaplan Meier survival analysis. There were 3 cases of catheter-related bacteremia per group. Logistic regression found that burns diagnosis and increased ICU stay significantly predicted colonization. CONCLUSION: IV administration sets can be used for 7 days in patients with short-term, antiseptic-coated CVCs. PMID- 15357157 TI - Predictors of nosocomial bloodstream infections among critically ill adult trauma patients. AB - OBJECTIVE: To identify the independent predictors of nosocomial bloodstream infections (BSIs) among critically ill adult trauma patients. DESIGN: A prospective, cohort design was used to study patients who met predetermined inclusion criteria. Basic descriptive and univariate statistical analyses were performed to identify unadjusted predictors. A forward stepwise multivariate logistic regression analysis was then conducted to identify independent predictors of nosocomial BSI. SETTING: Level I university-affiliated shock trauma center. PATIENTS: Three hundred sixty-one critically ill adult trauma patients, 55 of whom developed nosocomial BSIs (15.2%). RESULTS: Data analysis of 45 variables indicated that only 9 were independent predictors of nosocomial BSI: presence of a chest tube, use of immunosuppressive agents, presence of microbial resistance, length of stay, presence of preexisting infection, percentage change of serum albumin levels, patient disposition, transfusion of 10 or more units of blood, and number of central venous catheters (CVCs) for patients who had 4 or more. The classification index of the final regression model at a cut-off point of 0.5 had a specificity of 97.4%, a sensitivity of 60%, a positive predictive value of 76.7%, a negative predictive value of 93%, and an overall precision of 91%. CONCLUSION: In this study, only 9 variables were independent predictors of nosocomial BSI. Our findings are specific to critically ill adult trauma patients and should be interpreted within the context of this particular population. PMID- 15357158 TI - In vitro assessment of chlorhexidine gluconate-impregnated polyurethane foam antimicrobial dressing using zone of inhibition assays. AB - OBJECTIVE: To evaluate an antimicrobial dressing consisting of hydrophilic polyurethane foam with chlorhexidine gluconate for activity against several antibiotic-resistant clinical isolates as well as American Type Culture Collection reference strains using zone of inhibition assays. METHODS: Sterile foam samples with chlorhexidine gluconate and untreated controls were transferred onto inoculated agar plates. Plates were incubated at 35 degrees C to 37 degrees C for 24 hours and examined for zones of inhibition around the foam samples. RESULTS: Polyurethane foam with chlorhexidine gluconate showed antimicrobial activity in vitro against all of the challenge organisms including antibiotic resistant clinical isolates. CONCLUSION: The data from this in vitro study support the hypothesis that polyurethane foam with chlorhexidine gluconate has an antimicrobial effect against antibiotic-resistant Staphylococcus and Enterococcus species, as well as Candida species. PMID- 15357159 TI - Cost-benefit analysis of chlorhexidine gluconate dressing in the prevention of catheter-related bloodstream infections. AB - OBJECTIVES: To compare the costs with the benefits of using chlorhexidine gluconate dressings on central venous catheters and to determine the effectiveness of these dressings in reducing local infections and catheter related bloodstream infections (CRBSIs), costs, and mortality. DESIGN: Cost benefit analysis using randomized, controlled trial data on chlorhexidine dressing prevention of local infection and CRBSI, data on cost of chlorhexidine dressing versus standard treatment, data on averted cost of treating local infection and CRBSI, and data on mortality attributable to CRBSI. Decision analysis evaluated averted CRBSI treatment cost per patient resulting from chlorhexidine dressing use. Sensitivity analyses demonstrated net benefit of chlorhexidine dressing, varying baseline rate of CRBSI, incremental cost of treating CRBSI, and number of catheters, and evaluated mortality preventable through chlorhexidine dressing use, varying baseline rate of CRBSI, number of catheters, and mortality attributable to CRBSI. PATIENTS AND SETTING: Patients of all Philadelphia area hospitals and one Philadelphia academic medical center. RESULTS: Estimated potential annual U.S. net benefits from chlorhexidine dressing use ranged from $275 million to approximately $1.97 billion. Cost-benefit findings persisted in sensitivity analyses varying baseline rate of CRBSI, incremental cost of treating CRBSI, and overall number of catheters used. Preventable mortality analyses showed potential decreases of between 329 and 3,906 U.S. deaths annually as a result of nationwide use of chlorhexidine dressing. CONCLUSIONS: Chlorhexidine dressings would reduce costs, local infections and CRBSIs, and deaths. Use of chlorhexidine dressings should be considered to prevent infections among patients with catheters. PMID- 15357160 TI - Prevention of central venous catheter-related bloodstream infections using non technologic strategies. AB - OBJECTIVE: To evaluate the incidence of nosocomial bacteremias related to the use of non-impregnated central venous catheters (CVCs) when only non-technologic strategies were used to prevent them. DESIGN: This was a prospective study of infectious complications of CVCs placed in intensive care unit (ICU) patients from April 1997 to December 2001. SETTING: The medical-surgical ICU of a tertiary care, university-affiliated hospital in Argentina. METHODS: We studied all patients admitted to the ICU using non-impregnated CVCs. Maximal sterile barrier precautions (ie, use of cap, mask, sterile gown, sterile gloves, and large sterile drape), strict handwashing, preparation of the patients' skin with antiseptic solutions, insertion and management of catheters by trained personnel, and continuing quality improvement programs aimed at appropriate insertion and maintenance of catheters were employed. RESULTS: During the study period, 2,525 patients were admitted to the ICU. Eight hundred sixty-eight patients had 1,037 CVCs inserted. The number of CVC-related bloodstream infections (BSIs), acquired in the ICU, was 2.7 per 1,000 CVC-days (13 nosocomial CVC-related BSIs during 4,770 days of CVC use). Microorganisms isolated included methicillin-susceptible Staphylococcus aureus (n = 6), methicillin-resistant S. aureus (n = 2), coagulase negative methicillin-resistant Staphylococcus (n = 2), Escherichia coli (n = 1), Klebsiella pneumoniae (n = 1), and Enterobacter cloacae (n = 1). CONCLUSIONS: A low rate of catheter-related BSI was achieved without antimicrobial-impregnated catheters. The incidence of CVC-associated bacteremias corresponded to the 10th to 20th percentile range of the National Nosocomial Infections Surveillance System hospitals for the same type of ICU. PMID- 15357161 TI - An outbreak of methicillin-resistant Staphylococcus aureus infections related to central venous catheters for hemodialysis. AB - OBJECTIVES: To determine risk factors for hemodialysis catheter-related bloodstream infections (HCRBSIs) and investigate whether use of maximal sterile barrier precautions would prevent HCRBSIs. SETTING: Tertiary-care medical center hemodialysis unit. DESIGN: Open trial with historical comparison and case-control study of risk factors for HCRBSIs. METHODS: Prospective surveillance was used to compare HCRBSI rates for 1 year before and after implementation of maximal sterile barrier precautions. A case-control study compared 50 case-patients with HCRBSI with 51 randomly selected control-patients. RESULTS: The HCRBSI rate was 1.6% per 100 dialysis runs (CI95, 1.1%-2.3%) in the first year and 0.77% (CI95, 0.5%-1.1%) in the second year (P = .0106). The most frequent cause of HCRBSI was MRSA in the first year (15 of 32) and MSSA in the second year (13 of 18). Ten MRSA blood isolates in the first year were identical by PFGE. Diabetes mellitus was a risk factor for HCRBSI. Age, gender, site of hemodialysis central venous catheter (CVC), other underlying diseases, coma score, APACHE II score, serum albumin level, and cholesterol level were not associated with HCRBSI and did not change between the 2 years. Hospital stay was prolonged for case-patients (32.78 +/- 20.96 days) versus control-patients (22.75 +/- 17.33 days), but mortality did not differ. CONCLUSIONS: Use of maximal sterile barrier precautions during the insertion of CVCs reduced HCRBSIs in dialysis patients and seemed cost-effective. Diabetes mellitus was associated with HCRBSI. An outbreak of MRSA in the first year was likely caused by cross-infection via medical personnel. PMID- 15357162 TI - Influence of selective bowel decontamination on the organisms recovered during bacteremia in neutropenic patients. AB - OBJECTIVE: To assess the influence of prophylactic selective bowel decontamination (SBD) on the spectrum of microbes causing bloodstream infection (BSI). DESIGN: The microbes causing BSI in neutropenic patients of a hematologic ward (HW) and a bone marrow transplantation unit (BMTU), respectively, were compared by retrospective analysis of blood culture results from January 1996 to June 2003. SETTING: A 30-bed HW (no SBD) and a BMTU including a 7-bed normal care ward and an 8-bed intensive care unit (SBD used) of a 2,200-bed university teaching hospital. RESULTS: The overall incidences of bacteremia in the HW and the BMTU were similar (72.6 vs 70.6 episodes per 1,000 admissions; P = .8). Two hundred twenty episodes of BSI were recorded in 164 neutropenic patients of the HW and 153 episodes in 127 neutropenic patients of the BMTU. Enterobacteriaceae (OR, 3.14; CI95, 1.67-5.97; P= .0002) and Streptococcus species (OR, 2.04; CI95, 1.14-3.70; P = .015) were observed more frequently in HW patients and coagulase negative staphylococci more frequently in BMTU patients (OR, 0.15; CI95, 0.09 0.26; P < .00001). No statistically significant differences were found for gram negative nonfermentative bacilli (P = .53), Staphylococcus aureus (P = .21), Enterococcus species (P = .48), anaerobic bacteria (P = .1), or fungi (P = .50). CONCLUSIONS: SBD did not lead to a significant reduction in the incidence of bacteremia, but significant changes in microbes recovered from blood cultures were observed. SBD should be considered when empiric antimicrobial therapy is prescribed for suspected BSI. PMID- 15357163 TI - Application of data mining techniques to healthcare data. AB - A high-level introduction to data mining as it relates to surveillance of healthcare data is presented. Data mining is compared with traditional statistics, some advantages of automated data systems are identified, and some data mining strategies and algorithms are described. A concrete example illustrates steps involved in the data mining process, and three successful data mining applications in the healthcare arena are described. PMID- 15357164 TI - Povidone-iodine ointment and gauze dressings associated with reduced catheter related infection in seriously ill neurosurgical patients. AB - Povidone-iodine ointment and gauze covered by transparent dressings were compared with transparent dressings alone in historical controls (both changed twice weekly) in neurosurgical patients needing catheter placement for prolonged periods. Colonization and bloodstream infection were both reduced with the new method (P < .01 and P = .062, respectively). PMID- 15357165 TI - The case of Ms. R. PMID- 15357166 TI - Screening for lung cancer with computed tomography: an examination of the evidence. AB - Smokers and others who are at high risk for lung cancer could benefit from a screening test that would find the cancer at an early stage, when the most effective treatments are available. How far are we from such a test? PMID- 15357167 TI - Minimizing recurrences of genital herpes--a role for suppressive antiviral therapy. AB - Stress may contribute to recurrent episodes of genital herpes, which themselves cause more stress. minimizing the frequency of recurrences--or eliminating them altogether--may help to break the cycle. PMID- 15357168 TI - Anthrax exposure--stay alert, act swiftly. AB - Initiating appropriate antibiotic therapy as soon as anthrax infection is suspected is not only acceptable but is also justified by the outcomes of the 2001 anthrax cases. PMID- 15357169 TI - Preventing a fatal outcome Addison's disease. AB - Low serum sodium levels combined with decreased extracellular volume are your key to recognizing this potentially lethal but very treatable disease. PMID- 15357170 TI - Case of the month. MCP II and III erosions of the right hand. PMID- 15357171 TI - The Supreme Court lets HMOs off the hook. PMID- 15357172 TI - A physician's guide to Stark rules. PMID- 15357173 TI - It's never "just a test". PMID- 15357174 TI - Hiring staff. PMID- 15357175 TI - Have job seekers fill this out. PMID- 15357176 TI - My 3-step hiring test. PMID- 15357177 TI - Access fees worth the risk? What to charge? PMID- 15357178 TI - What about a malpractice surcharge. PMID- 15357179 TI - What not to document. PMID- 15357180 TI - Online compensation for target motion with scanned particle beams: simulation environment. AB - Target motion is one of the major limitations of each high precision radiation therapy. Using advanced active beam delivery techniques, such as the magnetic raster scanning system for particle irradiation, the interplay between time dependent beam and target position heavily distorts the applied dose distribution. This paper presents a simulation environment in which the time dependent effect of target motion on heavy-ion irradiation can be calculated with dynamically scanned ion beams. In an extension of the existing treatment planning software for ion irradiation of static targets (TRiP) at GSI, the expected dose distribution is calculated as the sum of several sub-distributions for single target motion states. To investigate active compensation for target motion by adapting the position of the therapeutic beam during irradiation, the planned beam positions can be altered during the calculation. Applying realistic parameters to the planned motion-compensation methods at GSI, the effect of target motion on the expected dose uniformity can be simulated for different target configurations and motion conditions. For the dynamic dose calculation, experimentally measured profiles of the beam extraction in time were used. Initial simulations show the feasibility and consistency of an active motion compensation with the magnetic scanning system and reveal some strategies to improve the dose homogeneity inside the moving target. The simulation environment presented here provides an effective means for evaluating the dose distribution for a moving target volume with and without motion compensation. It contributes a substantial basis for the experimental research on the irradiation of moving target volumes with scanned ion beams at GSI which will be presented in upcoming papers. PMID- 15357181 TI - Statistical pixelwise inference models for planar data analysis: an application to gamma-camera uniformity monitoring. AB - In this paper two tests based on statistical models are presented and used to assess, quantify and provide positional information of the existence of bias and/or variations between planar images acquired at different times but under similar conditions. In the first test a linear regression model is fitted to the data in a pixelwise fashion, using three mathematical operators. In the second test a comparison using z-scoring is used based on the assumption that Poisson statistics are valid. For both tests the underlying assumptions are as simple and few as possible. The results are presented as parametric maps of either the three operators or the z-score. The z-score maps can then be thresholded to show the parts of the images which demonstrate change. Three different thresholding methods (naive, adaptive and multiple) are presented: together they cover almost all the needs for separating the signal from the background in the z-score maps. Where the expected size of the signal is known or can be estimated, a spatial correction technique (referred to as the reef correction) can be applied. These tests were applied to flood images used for the quality control of gamma camera uniformity. Simulated data were used to check the validity of the methods. Real data were acquired from four different cameras from two different institutions using a variety of acquisition parameters. The regression model found the bias in all five simulated cases and it also found patterns of unstable regions in real data where visual inspection of the flood images did not show any problems. In comparison the z-map revealed the differences in the simulated images from as low as 1.8 standard deviations from the mean, corresponding to a differential uniformity of 2.2% over the central field of view. In all cases studied, the reef correction increased significantly the sensitivity of the method and in most cases the specificity as well. The two proposed tests can be used either separately or in combination and are capable of showing trends and/or the magnitude of difference between images acquired under similar conditions with high positional and statistical precision. In addition to gamma camera quality control, they could be applied to any pair (or set) of registered planar images to detect subtle changes, e.g. a set of scintigrams or conventional radiographs of a patient before, during and after treatment. PMID- 15357182 TI - Fast free-form deformable registration via calculus of variations. AB - In this paper, we present a fully automatic, fast and accurate deformable registration technique. This technique deals with free-form deformation. It minimizes an energy functional that combines both similarity and smoothness measures. By using calculus of variations, the minimization problem was represented as a set of nonlinear elliptic partial differential equations (PDEs). A Gauss-Seidel finite difference scheme is used to iteratively solve the PDE. The registration is refined by a multi-resolution approach. The whole process is fully automatic. It takes less than 3 min to register two three-dimensional (3D) image sets of size 256 x 256 x 61 using a single 933 MHz personal computer. Extensive experiments are presented. These experiments include simulations, phantom studies and clinical image studies. Experimental results show that our model and algorithm are suited for registration of temporal images of a deformable body. The registration of inspiration and expiration phases of the lung images shows that the method is able to deal with large deformations. When applied to the daily CT images of a prostate patient, the results show that registration based on iterative refinement of displacement field is appropriate to describe the local deformations in the prostate and the rectum. Similarity measures improved significantly after the registration. The target application of this paper is for radiotherapy treatment planning and evaluation that incorporates internal organ deformation throughout the course of radiation therapy. The registration method could also be equally applied in diagnostic radiology. PMID- 15357183 TI - Exact filtered backprojection reconstruction for dynamic pitch helical cone beam computed tomography. AB - We present an exact filtered backprojection reconstruction formula for helical cone beam computed tomography in which the pitch of the helix varies with time. We prove that the resulting algorithm, which is functionally identical to the constant pitch case, provides exact reconstruction provided that the projection of the helix onto the detector forms convex boundaries and that PI lines are unique. Furthermore, we demonstrate that both of these conditions are satisfied provided the sum of the translational velocity and the derivative of the translational acceleration does not change sign. As a special case, we show that gantry tilt can also be handled by our dynamic pitch formula. Simulation results demonstrate the resulting algorithm. PMID- 15357184 TI - Dynamic infrared imaging for the detection of malignancy. AB - The potential for malignancy detection using dynamic infrared imaging (DIRI) has been investigated in an animal model of human malignancy. Malignancy was apparent in images formed at the vasomotor and cardiogenic frequencies of tumour bearing mice. The observation of malignancy was removed by the administration of an agent that blocks vasodilation caused by nitric oxide (NO). Image patterns similar to those that characterize malignancy could be mimicked in normal mice using an NO producing agent. Apparently DIRI allows for cancer detection in this model through vasodilation caused by malignancy generated NO. Dynamic infrared detection of vasomotor and cardiogenic surface perfusion was validated in human subjects by a comparison with laser Doppler flowmetry (LDF). Dynamic infrared imaging technology was then applied to breast cancer detection. It is shown that dynamic infrared images formed at the vasomotor and cardiogenic frequencies of the normal and malignant breast have image pattern differences, which may allow for breast cancer detection. PMID- 15357185 TI - Photoacoustic imaging with deconvolution algorithm. AB - The impulse response of the ultrasonic transducer used for detection is crucial for photoacoustic imaging with high resolution. We demonstrate a reconstruction method that allows the optical absorption distribution of a sample to be reconstructed without knowing the impulse response of the ultrasonic transducer. A convolution relationship between photoacoustic signals measured by an ultrasound transducer and optical absorption distribution is developed. Based on this theory, the projection of the optical absorption distribution of a sample can be obtained directly by deconvolving the recorded PA signal originating from a point source out of that from the sample. And a modified filtered back projection algorithm is used to reconstruct the optical absorption distribution. We constructed a photoacoustic imaging system to validate the reconstruction method and the experimental results demonstrated that the reconstructed images agreed well with the original phantom samples. The spatial resolution of the system reaches 0.3 mm. PMID- 15357186 TI - Study of the point spread function (PSF) for 123I SPECT imaging using Monte Carlo simulation. AB - The iterative reconstruction algorithms employed in brain single-photon emission computed tomography (SPECT) allow some quantitative parameters of the image to be improved. These algorithms require accurate modelling of the so-called point spread function (PSF). Nowadays, most in vivo neurotransmitter SPECT studies employ pharmaceuticals radiolabelled with 123I. In addition to an intense line at 159 keV, the decay scheme of this radioisotope includes some higher energy gammas which may have a non-negligible contribution to the PSF. The aim of this work is to study this contribution for two low-energy high-resolution collimator configurations, namely, the parallel and the fan beam. The transport of radiation through the material system is simulated with the Monte Carlo code PENELOPE. We have developed a main program that deals with the intricacies associated with tracking photon trajectories through the geometry of the collimator and detection systems. The simulated PSFs are partly validated with a set of experimental measurements that use the 511 keV annihilation photons emitted by a 18F source. Sensitivity and spatial resolution have been studied, showing that a significant fraction of the detection events in the energy window centred at 159 keV (up to approximately 49% for the parallel collimator) are originated by higher energy gamma rays, which contribute to the spatial profile of the PSF mostly outside the 'geometrical' region dominated by the low-energy photons. Therefore, these high energy counts are to be considered as noise, a fact that should be taken into account when modelling PSFs for reconstruction algorithms. We also show that the fan beam collimator gives higher signal-to-noise ratios than the parallel collimator for all the source positions analysed. PMID- 15357187 TI - A comparison of step-and-shoot leaf sequencing algorithms that eliminate tongue and-groove effects. AB - The performances of three recently published leaf sequencing algorithms for step and-shoot intensity-modulated radiation therapy delivery that eliminates tongue and-groove underdosage are evaluated. Proofs are given to show that the algorithm of Que et al (2004 Phys. Med. Biol. 49 399-405) generates leaf sequences free of tongue-and-groove underdosage and interdigitation. However, the total beam-on times could be up to n times those of the sequences generated by the algorithms of Kamath et al (2004 Phys. Med. Biol. 49 N7-N19), which are optimal in beam-on time for unidirectional leaf movement under the same constraints, where n is the total number of involved leaf pairs. Using 19 clinical fluence matrices and 100000 randomly generated 15 x 15 matrices, the average monitor units and number of segments of the leaf sequences generated using the algorithm of Que et al are about two to four times those generated by the algorithm of Kamath et al. PMID- 15357188 TI - Radiation-induced statistical uncertainty in the threshold voltage measurement of MOSFET dosimeters. AB - The results of a recent study on the limiting uncertainties in the measurement of photon radiation dose with MOSFET dosimeters are reported. The statistical uncertainty in dose measurement from a single device has been measured before and after irradiation. The resulting increase in 1/f noise with radiation dose has been investigated via various analytical models. The limit of uncertainty in the ubiquitous linear trend of threshold voltage with dose has been measured and compared to two nonlinear models. Inter-device uncertainty has been investigated in a group of 40 devices, and preliminary evidence for kurtosis and skewness in the distributions for devices without external bias has been observed. PMID- 15357189 TI - An evaluation of the AAPM-TG43 dosimetry protocol for I-125 brachytherapy seed. AB - The EGSnrc Monte Carlo system has been used to calculate the dose distributions from 125I radioactive seeds (model 6711). The results showed that the agreement between EGSnrc and the American Association of Physicists in Medicine Task Group Report 43 (AAPM-TG43) dosimetry protocol is generally within +/-15% for radial distances less than 1.0 cm in both the transverse axis and longitudinal axis of the source. For radial distances between 1.0 and 2.5 cm the agreement between Monte Carlo simulations and the AAPM-TG43 dosimetry protocol is within +/-20%. In the longitudinal axis of the source the difference between Monte Carlo simulations and the AAPM-TG43 dosimetry is up to 40% for radial distances greater than 2.5 cm. The agreement between the EGSnrc/Monte Carlo simulation and the AAPM TG43 dosimetry protocol improved significantly when recently published data of the anisotropic function were implemented (Weaver 1998 Med. Phys. 25 2271-8). The difference between Monte Carlo simulations and the AAPM-TG43 dosimetry protocol is not more than +/-10% in the transverse axis of the source up to a radius of 2.5 cm. The EGSnrc Monte Carlo simulation and the AAPM-TG43 with the Weaver anisotropic data were also used to investigate the differences in the dose distribution caused by small differences in the construction of individual seeds (Sloboda and Menon 2000 Med. Phys. 27 1789-99). The results show that a change in length of the silver rod containing the 125I radioactive material of 0.14 mm does not affect the dose distribution significantly in the transverse and longitudinal axes but a change of 0.13 mm in the thickness of the welded end of the encapsulation affected the dose significantly in the longitudinal axis of the source. PMID- 15357190 TI - Dose perturbation effects in prostate seed implant brachytherapy with I-125. AB - EGSnrc Monte Carlo simulation was used to investigate dose perturbation effects in prostate seed implant brachytherapy using 125I radioactive seeds used in implant brachytherapy. Dose perturbation effects resulting from the seed mutual attenuation in a prostate seed implant consisting of 27 seeds were investigated. The results showed that for 125I seeds implanted into the prostate at 1.00 cm, 0.75 cm and 0.50 cm apart (uniform spacing), the dose perturbation effects are up to 10%. The volume of the target occupied by the 10% dose difference between the full Monte Carlo simulation and the single seed superposition model decreases with increasing seed spacing. Despite the differences between the Monte Carlo simulation and the simple superposition, there was no significant change in the dose volume histogram for 1 cm and 0.75 cm seed spacing. However, there was a significant change in the dose volume histogram when the seed spacing was 0.5 cm. An analysis of the external volume index (EI), coverage index (CI) and homogeneity index (HI) also showed that there is no difference in these indexes for the 1.00 cm and 0.75 cm seed spacing between the simple superposition model and the full Monte Carlo simulation. Compared to the full Monte Carlo simulations, the simple superposition model overestimated EI, CI and HI by 7%, 5% and 4% respectively for the 0.50 cm seed spacing. PMID- 15357191 TI - Range verification system using positron emitting beams for heavy-ion radiotherapy. AB - It is desirable to reduce range ambiguities in treatment planning for making full use of the major advantage of heavy-ion radiotherapy, that is, good dose localization. A range verification system using positron emitting beams has been developed to verify the ranges in patients directly. The performance of the system was evaluated in beam experiments to confirm the designed properties. It was shown that a 10C beam could be used as a probing beam for range verification when measuring beam properties. Parametric measurements indicated the beam size and the momentum acceptance and the target volume did not influence range verification significantly. It was found that the range could be measured within an analysis uncertainty of +/-0.3 mm under the condition of 2.7 x 10(5) particle irradiation, corresponding to a peak dose of 96 mGyE (gray-equivalent dose), in a 150 mm diameter spherical polymethyl methacrylate phantom which simulated a human head. PMID- 15357192 TI - Light dosimetry for multiple cylindrical diffusing sources for use in photodynamic therapy. AB - Since prostatic carcinoma is usually multifocal within the prostate, effective photodynamic therapy (PDT) of prostatic carcinoma is expected to require the photochemical destruction of the entire organ. Accurate light dosimetry will be essential to avoid damage to proximal sensitive tissue such as the rectum. The prostate will be illuminated using interstitial cylindrical fibreoptic light sources and, because of the limited transparency of prostate tissue, these sources will be mounted in a parallel array analogous to the source array used in brachytherapy. Both source spacing and the light delivered to each source will control light dosimetry from a parallel array of fibreoptic sources implanted into tissue. Clinical PDT will require dose planning in order to determine the position and illumination of each source prior to treatment, but unfortunately few methods of predicting light fluence from cylindrical interstitial sources currently exist. In this paper, a novel light fluence model is used to predict tissue transillumination resulting from cylindrical interstitial sources. The cylindrical source is modelled as a finite array of infinitesimal small sources using Christian Huygens' famous single-slit diffraction model. We show that this source model when combined with a robust derivation of fluence in a spherical geometry using diffusion theory, accurately predicts fluence levels from a single cylindrical source in a variety of media. This method is found to retain its accuracy near the sources. With a simple extension, this fluence model is used to predict the light fluence levels from an array of three sources and the predicted fluence is found to compare favourably with experimental data. PMID- 15357193 TI - Transperineal in vivo fluence-rate dosimetry in the canine prostate during SnET2 mediated PDT. AB - Advances in photodynamic therapy (PDT) treatment for prostate cancer can be achieved either by improving selectivity of the photosensitizer towards prostate gland tissue or improving the dosimetry by means of individualized treatment planning using currently available photosensitizers. The latter approach requires the ability to measure, among other parameters, the fluence rate at different positions within the prostate and the ability to derive the tissue optical properties. Here fibre optic probes are presented capable of measuring the fluence rate throughout large tissue volumes and a method to derive the tissue optical properties for different volumes of the prostate. The responsivity of the sensors is sufficient to detect a fluence rate of 0.1 mW cm(-2). The effective attenuation coefficient in the canine prostate at 660 nm is higher at the capsule (2.15+/-0.19 cm(-1)) than in proximity of the urethra (1.84+/-0.36 cm(-1)). Significant spatial and temporal intra- and inter-canine variability in the tissue optical properties was noted, highlighting the need for individualized monitoring of the fluence rate for improved dosimetry. PMID- 15357194 TI - Inherent smoothness of intensity patterns for intensity modulated radiation therapy generated by simultaneous projection algorithms. AB - The efficient delivery of intensity modulated radiation therapy (IMRT) depends on finding optimized beam intensity patterns that produce dose distributions, which meet given constraints for the tumour as well as any critical organs to be spared. Many optimization algorithms that are used for beamlet-based inverse planning are susceptible to large variations of neighbouring intensities. Accurately delivering an intensity pattern with a large number of extrema can prove impossible given the mechanical limitations of standard multileaf collimator (MLC) delivery systems. In this study, we apply Cimmino's simultaneous projection algorithm to the beamlet-based inverse planning problem, modelled mathematically as a system of linear inequalities. We show that using this method allows us to arrive at a smoother intensity pattern. Including nonlinear terms in the simultaneous projection algorithm to deal with dose-volume histogram (DVH) constraints does not compromise this property from our experimental observation. The smoothness properties are compared with those from other optimization algorithms which include simulated annealing and the gradient descent method. The simultaneous property of these algorithms is ideally suited to parallel computing technologies. PMID- 15357195 TI - Influence of multiple scattering and energy loss straggling on the absorbed dose distributions of therapeutic light ion beams: I. Analytical pencil beam model. AB - The lateral and longitudinal distributions of absorbed dose of broad and narrow light ion beams in water are investigated. An analytical algorithm based on the generalized Fermi-Eyges theory is developed and used to calculate the effects of multiple scattering and range straggling on the dose distribution of light ion beams in water. A first-order Gaussian multiple scattering and energy loss straggling approach is generally sufficiently accurate for describing the lateral and longitudinal spread of the Bragg peak and the associated energy deposition distribution of therapeutic light ion beams at ranges of clinical interest. Nuclear reactions are not taken into account in this study. The analytical algorithm given in the present study allows an accurate description of the radial spread and the range straggling of light ions traversing matter. A verification of this approach by comparing with experimental data, Monte Carlo methods and other analytical techniques will be presented in a forthcoming paper. PMID- 15357196 TI - A blood-oxygenation-dependent increase in blood viscosity due to a static magnetic field. AB - As the magnetic field of widely used MR scanners is one of the strongest magnetic fields to which people are exposed, the biological influence of the static magnetic field of MR scanners is of great concern. One magnetic interaction in biological subjects is the magnetic torque on the magnetic moment induced by biomagnetic substances. The red blood cell is a major biomagnetic substance, and the blood flow may be influenced by the magnetic field. However, the underlying mechanisms have been poorly understood. To examine the mechanisms of the magnetic influence on blood viscosity, we measured the time for blood to fall through a glass capillary inside and outside a 1.5 T MR scanner. Our in vitro results showed that the blood viscosity significantly increased in a 1.5 T MR scanner, and also clarified the mechanism of the interaction between red blood cells and the external magnetic field. Notably, the blood viscosity increased depending on blood oxygenation and the shear rate of the blood flow. Thus, our findings suggest that even a 1.5 T magnetic field may modulate blood flow. PMID- 15357197 TI - Registration of electronic portal images for patient set-up verification. AB - Images acquired from an electronic portal imaging device are aligned with digitally reconstructed radiographs (DRRs) or other portal images to verify patient positioning during radiation therapy. Most of the currently available computer aided registration methods are based on the manual placement of corresponding landmarks. The purpose of the paper is twofold: (a) the establishment of a methodology for patient set-up verification during radiotherapy based on the registration of electronic portal images, and (b) the evaluation of the proposed methodology in a clinical environment. The estimation of set-up errors, using the proposed methodology, can be accomplished by matching the portal image of the current fraction of the treatment with the portal image of the baseline treatment (reference portal image) using a nearly automated technique. The proposed registration method is tested on a number of phantom data as well as on data from four patients. The phantom data included portal images that corresponded to various positions of the phantom on the treatment couch. For each patient, a set of 30 portal images was used. For the phantom data (for both transverse and lateral portal images), the maximum absolute deviations of the translational shifts were within 1.5 mm, whereas the in-plane rotation angle error was less than 0.5 degrees. The two-way Anova revealed no statistical significant variability both within observer and between-observer measurements (P > 0.05). For the patient data, the mean values obtained with manual and the proposed registration methods were within 0.5 mm. In conclusion, the proposed registration method has been incorporated within a system, called ESTERR-PRO. Its image registration capability achieves high accuracy and both intra- and inter user reproducibility. The system is fully operational within the Radiotherapy Department of 'HYGEIA' Hospital in Athens and it could be easily installed in any other clinical environment since it requires standardized hardware specifications and minimal human intervention. PMID- 15357198 TI - K-edge digital subtraction imaging based on a dichromatic and compact x-ray source. AB - This work proposes a compact dichromatic imaging system for the application of the K-edge digital subtraction technique based on a conventional x-ray tube and a monochromator system. A quasi-monochromatic x-ray beam at the energy of iodine K edge is produced by Bragg diffraction on a mosaic crystal. Two thin adjacent beams with energies that bracket the K-edge discontinuity are obtained from the diffracted beam by means of a proper collimation system. They are then detected using an array of Si detectors. A home-made phantom is used to study the image quality as a function of iodine concentration. Signal and signal-to-noise ratio analysis has also been performed. The results are compared with theoretical expectations. PMID- 15357199 TI - Choice of the regularization parameter for perfusion quantification with MRI. AB - Truncated singular value decomposition (TSVD) is an effective method for the deconvolution of dynamic contrast enhanced (DCE) MRI. Two robust methods for the selection of the truncation threshold on a pixel-by-pixel basis--generalized cross validation (GCV) and the L-curve criterion (LCC)--were optimized and compared to paradigms in the literature. GCV and LCC were found to perform optimally when applied with a smooth version of TSVD, known as standard form Tikhonov regularization (SFTR). The methods lead to improvements in the estimate of the residue function and of its maximum, and converge properly with SNR. The oscillations typically observed in the solution vanish entirely, and perfusion is more accurately estimated at small mean transit times. This results in improved image contrast and increased sensitivity to perfusion abnormalities, at the cost of 1-2 min in calculation time and hyperintense clusters in the image. Preliminary experience with clinical data suggests that the latter problem can be resolved using spatial continuity and/or hybrid thresholding methods. In the simulations GCV and LCC are equivalent in terms of performance, but GCV thresholding is faster. PMID- 15357201 TI - Optimization of standard patient radiographic images for chest, skull and pelvis exams in conventional x-ray equipment. AB - Optimized radiographic techniques for clinical images of chest, skull and pelvis using conventional single-phase, three-phase and high-frequency x-ray units for a standard patient have been developed. Optimization of image contrast and optical density was obtained by using a homogeneous phantom (PEP) and an Anderson Rando anthropomorphic phantom. Image quality was evaluated by nine radiologists in independent analyses, leading to the choice of the optimized technique. A course of action to implement and validate these techniques in other radiographic systems has also been introduced. A realistic-analytic phantom (RAP) was constructed to certify the validation process. The optimized radiographic technique was implemented in the routine of our home hospital radiodiagnostic routine, enabling a reduction in patient doses around 25, 14 and 72%, respectively, for chest, skull and pelvis exams when compared with the previously used techniques. In addition, a corresponding reduction in the x-ray tube load of 68, 14 and 62% for the respective mentioned exams has been observed. In conclusion, implemented optimal techniques can lead to a reduction in the rate of film rejection, thus contributing to a better risk-benefit relationship for the patient and cost-benefit for the radiodiagnostic facility. PMID- 15357200 TI - Haemodynamic resistance model of monochorionic twin pregnancies complicated by acardiac twinning. AB - An acardiac twin is a severely malformed monochorionic twin fetus that lacks most organs, particularly a heart. It grows during pregnancy, because it is perfused by its developmentally normal co-twin (called the pump twin) via a set of placental arterioarterial and venovenous anastomoses. The pump twin dies intrauterine or neonatally in about 50% of the cases due to congestive heart failure, polyhydramnios and prematurity. Because the pathophysiology of this pregnancy is currently incompletely understood, we modified our previous haemodynamic model of monochorionic twins connected by placental vascular anastomoses to include the analysis of acardiac twin pregnancies. We incorporated the fetoplacental circulation as a resistance circuit and used the fetal umbilical flow that perfuses the body to define fetal growth, rather than the placental flow as done previously. Using this modified model, we predicted that the pump twin has excess blood volume and increased mean arterial blood pressure compared to those in the acardiac twin. Placental perfusion of the acardiac twin is significantly reduced compared to normal, as a consequence of an increased venous pressure, possibly implying reduced acardiac placental growth. In conclusion, the haemodynamic analysis may contribute to an increased knowledge of the pathophysiologic consequences of an acardiac body mass for the pump twin. PMID- 15357202 TI - The specific uptake size index for quantifying radiopharmaceutical uptake. AB - Quantitative indices of radionuclide uptake in an object of interest provide a useful adjunct to qualitative interpretation in the diagnostic application of radionuclide imaging. This note describes a new measure of total uptake of an organ, the specific uptake size index (SUSI). It can either be related in absolute terms to the total activity injected or to the specific activity in a reference region. As it depends on the total activity in the object, the value obtained will not depend on the resolution of the imaging process, as is the case with some other similar quantitative indices. This has been demonstrated in an experiment using simulated images. The application of the index to quantification of dopamine receptor SPECT imaging and parathyroid-thyroid subtraction planar scintigraphy is described. The index is considered to be of potential value in reducing variation in quantitative assessment of uptake in objects with applications in all areas of radionuclide imaging. PMID- 15357203 TI - MCDE: a new Monte Carlo dose engine for IMRT. AB - A new accurate Monte Carlo code for IMRT dose computations, MCDE (Monte Carlo dose engine), is introduced. MCDE is based on BEAMnrc/DOSXYZnrc and consequently the accurate EGSnrc electron transport. DOSXYZnrc is reprogrammed as a component module for BEAMnrc. In this way both codes are interconnected elegantly, while maintaining the BEAM structure and only minimal changes to BEAMnrc.mortran are necessary. The treatment head of the Elekta SLiplus linear accelerator is modelled in detail. CT grids consisting of up to 200 slices of 512 x 512 voxels can be introduced and up to 100 beams can be handled simultaneously. The beams and CT data are imported from the treatment planning system GRATIS via a DICOM interface. To enable the handling of up to 50 x 10(6) voxels the system was programmed in Fortran95 to enable dynamic memory management. All region-dependent arrays (dose, statistics, transport arrays) were redefined. A scoring grid was introduced and superimposed on the geometry grid, to be able to limit the number of scoring voxels. The whole system uses approximately 200 MB of RAM and runs on a PC cluster consisting of 38 1.0 GHz processors. A set of in-house made scripts handle the parallellization and the centralization of the Monte Carlo calculations on a server. As an illustration of MCDE, a clinical example is discussed and compared with collapsed cone convolution calculations. At present, the system is still rather slow and is intended to be a tool for reliable verification of IMRT treatment planning in the case of the presence of tissue inhomogeneities such as air cavities. PMID- 15357204 TI - Photonuclear isotope characterization of a Siemens KDS 18 MV linac head. AB - Induced activity due to photonuclear reactions produced in the vacuum window of the accelerating wave-guide, the x-ray target and the beam-flattening filter of an 18 MV Siemens KDS linac has been studied. Measurements were performed using a high-purity portable germanium detector. Radioisotopes such as 196Au, 57Co, 60Co and other traces were detected one week after the last clinical use of the linac. PMID- 15357205 TI - Reduction of ring artefacts in high resolution micro-CT reconstructions. AB - High resolution micro-CT images are often corrupted by ring artefacts, prohibiting quantitative analysis and hampering post processing. Removing or at least significantly reducing such artefacts is indispensable. However, since micro-CT systems are pushed to the extremes in the quest for the ultimate spatial resolution, ring artefacts can hardly be avoided. Moreover, as opposed to clinical CT systems, conventional correction schemes such as flat-field correction do not lead to satisfactory results. Therefore, in this note a simple but efficient and fast post processing method is proposed that effectively reduces ring artefacts in reconstructed micro-CT images. PMID- 15357206 TI - Comment on the modified Beer-Lambert law for scattering media. AB - We present a concise overview of the modified Beer-Lambert law, which has been extensively used in the literature of near-infrared spectroscopy (NIRS) of scattering media. In particular, we discuss one form of the modified Beer-Lambert law that is commonly found in the literature and that is not strictly correct. However, this incorrect form of the modified Beer-Lambert law still leads to the correct expression for the changes in the continuous wave optical signal associated with changes in the absorption coefficient of the investigated medium. Here we propose a notation for the modified Beer-Lambert law that keeps the typical form commonly found in the literature without introducing any incorrect assumptions. PMID- 15357207 TI - The determinants of Chagas disease: connecting parasite and host genetics. AB - As a consequence of infection by Trypanosoma cruzi, 30% of victims may develop chronic Chagas disease, which presents a spectrum of pathology including cardiomyopathy, megacolon and megaesophagus. The outcome of infection in a particular individual is the result of a set of complex interactions among the host genetic background, environmental and social factors, and the genetic composition of the parasite, all of which can be complicated by mixed infections and re-infections. Initially we consider what is known about the genetic structure and biological properties of the protozoan. Currently, six distinct subgroups have been characterized by different combinations of four distinct genotypic classes. The recent demonstration of genetic exchange via non-meiotic cell fusion illustrates a mechanism by which maintained heterogeneous polyploidy may have been generated in these parasites. Subsequently, we consider factors in humans and in experimental mouse-infection and tissue culture models that have contributed to our understanding of the host's susceptibility or resistance to disease. Identification of the direct players in host-pathogen interactions at the establishment and chronic phases of the disease is perhaps the best hope of a clinical handle for treatment. At some point in the future, these disparate areas of study will have to come together. It is to be hoped that this scientific fusion will result in better prognosis and treatment of Chagas disease. PMID- 15357208 TI - The molecular control of antigenic variation in Trypanosoma brucei. AB - The African trypanosome, Trypanosoma brucei, is a protozoan that causes sleeping sickness in humans and N'gana in livestock. These flagellated parasites are directly exposed to immune defences as they circulate in the mammalian host bloodstream but they maintain persistent infections by undergoing antigenic variation. Central to this process is mono-allelic transcription and switching of the expressed variant-surface glycoprotein (VSG) gene which encodes the vast majority of their dense surface coat. The active telomeric VSG is transcribed by RNA polymerase I in an 'expression site body' (ESB) while transcription attenuation occurs at 'inactive' telomeres. Here, I review what is known about the molecular mechanisms involved in achieving antigenic variation and outline how we intend to exploit genome sequence and new tools, particularly RNA interference, to identify and characterise factors required for VSG regulation. PMID- 15357209 TI - Genetic regulation of protein synthesis in trypanosomes. AB - It is becoming increasingly clear that parasitic protozoa remain a scourge to humans in the 21st century. The trypanosomes are a diverse group of insect transmitted parasites that wiggle their way through multiple life cycle stages as they destroy human lives. Exquisitely detailed studies of these organisms reveal basic differences in gene expression that separate these single celled eukaryotes from multicellular eukaryotic organisms and have suggested numerous potential drug targets. PMID- 15357210 TI - cAMP signalling in the kinetoplastid protozoa. AB - Several species of kinetoplastid protozoa cause major human infectious diseases. Trypanosoma cruzi is responsible for the fatal Chagas disease in large parts of South America, the various species of Leishmania cause a number of different human diseases with millions of patients world-wide, and the African trypanosome Trypanosoma brucei is the agent of human sleeping sickness, a disastrously re emerging epidemic of fatal infections in Sub-Saharan Africa. Chemotherapy of all of these infections is in a very unsatisfactory state. cAMP signalling pathways in humans have provided interesting drug targets for a number of clinical conditions, from asthma to impotency. Similarly, cAMP signalling in kinetoplastids might offer useful targets for the development of novel antiparasitic drugs, which makes their exploration an urgent need. Current knowledge suggests that cAMP signalling proceeds along very similar pathways in all kinetoplastid pathogens (T. cruzi, the Leishmanias and T. brucei). Their adenylyl cyclases are structurally very different from the human enzymes and appear to function as enzyme-linked cell surface receptors. They might represent the major sensory apparatus of the kinetoplastids, guiding much of their environmental sensing and host/parasite interaction. The cAMP-specific phosphodiesterases of the kinetoplastids are rather similar to those of human cells and might function in similar ways. Essentially nothing is known on downstream effectors of cAMP in the kinetoplastids. Homologues of protein kinase A and its regulatory subunits have been identified, but their biochemical properties seem to be disctinct from that of mammalian protein kinase A. PMID- 15357211 TI - New insights into the developmental biology and transmission mechanisms of Leishmania. AB - Leishmania alternates between two main morphological forms in its life cycle: intracellular amastigotes in the mammalian host and motile promastigotes in the sandfly vector. Several different forms of promastigote can be recognised in sandfly infections. The first promastigote forms, which are found in the sandfly in the bloodmeal phase, are multiplicative procyclic promastigotes. These differentiate into nectomonad promastigotes, which are a non-dividing migratory stage moving from the posterior to the anterior midgut. When nectomonad promastigotes arrive at the anterior midgut they differentiate into leptomonad forms, a newly named life cycle stage, which resume replication. Leptomonad promastigotes, which are found in the anterior midgut, are the developmental precursors of the metacyclic promastigotes, the mammal-infective stages. Leptomonad forms also produce promastigote secretory gel, a substance that plays a key role in transmission by forming a physical obstruction in the gut, forcing the sandfly to regurgitate metacyclic promastigotes during bloodfeeding. PMID- 15357212 TI - The application of gene expression microarray technology to kinetoplastid research. AB - Protozoan parasites in the order Kinetoplastida cause severe disease primarily in tropical and subtropical areas. Vaccines to control these diseases have shown some promise, but none are in active clinical use. Drug treatments are available for all of the acute infections, but the emergence of resistance and an unresponsive chronic phase are current problems. Rapid advances in genomic technology open the possibility of discovering new genes that can contribute to vaccine initiatives or serve as targets for development of new drugs. The DNA microarray is a genomic technology, which is being applied to new gene discovery in kinetoplastid parasites. Both cDNA and genomic microarrays for Leishmania major have identified a number of new genes that are expressed in a stage specific fashion and preliminary results from a L. donovani genomic microarray also demonstrated new gene discovery. A microarray of Trypanosoma brucei genomic fragments identified new genes whose expression differs between the insect borne stage and the human infectious stage of the parasite. The next few years, building on this foundational work, should witness the most exciting stage as microarrays are applied to questions such as the basis of drug resistance, post kala azar dermal leishmaniasis, the regulation of differentiation to infectious stages, linking coordinately regulated pathways of genes and development of genetically defined parasites that may have potential as live attenuated vaccines. PMID- 15357213 TI - The structure and replication of kinetoplast DNA. AB - Kinetoplast DNA (kDNA), the mitochondrial DNA of flagellated protozoa of the order Kinetoplastida, is unique in its structure, function and mode of replication. It consists of few dozen maxicircles, encoding typical mitochondrial proteins and ribosomal RNA, and several thousands minicircles, encoding guide RNA molecules that function in the editing of maxicircles mRNA transcripts. kDNA minicircles and maxicircles in the parasitic species of the family Trypanosomatidae are topologically linked, forming a two dimensional fishnet-type DNA catenane. Studies of early branching free-living and parasitic species of the Bodonidae family revealed various other forms of this remarkable DNA structure and suggested the evolution of kDNA from unlinked DNA circles and covalently linked concatamers into a giant topological catenane. The replication of kDNA occurs during nuclear S phase and includes the duplication of free detached minicircles and catenated maxicircle and the generation of two progeny kDNA networks that segregate upon cell division. Recent reports of sequence elements and specific proteins that regulate the periodic expression of replication proteins advanced our understanding of the mechanisms that regulate the temporal link between mitochondrial and nuclear DNA synthesis in trypanosomatids. Studies on kDNA replication enzymes and binding proteins revealed their remarkable organization in clusters at defined sites flanking the kDNA disk, in correlation with the progress in the cell cycle and the process of kDNA replication. In this review I describe the recent advances in the study of kDNA and discuss some of the major challenges in deciphering the structure, replication and segregation of this remarkable DNA structure. PMID- 15357214 TI - Surface determinants of Leishmania parasites and their role in infectivity in the mammalian host. AB - Leishmania are intracellular protozoan parasites that reside primarily in host mononuclear phagocytes. Infection of host macrophages is initiated by infective promastigote stages and perpetuated by an obligate intracellular amastigote stage. Studies undertaken over the last decade have shown that the composition of the complex surface glycocalyx of these stages (comprising lipophosphoglycan, GPI anchored glycoproteins, proteophosphoglycans and free GPI glycolipids) changes dramatically as promastigotes differentiate into amastigotes. Marked stage specific changes also occur in the expression of other plasma membrane components, including type-1, polytopic and peripheral membrane proteins, reflecting the distinct microbicidal responses and nutritional environments encountered by these stages. More recently, a number of Leishmania mutants lacking single or multiple surface components have been generated. While some of these mutants are less virulent than wild type parasites, many of these mutants exhibit only mild or no loss of virulence. These studies suggest that, 1) the major surface glycocalyx components of the promastigote stage (i.e. LPG, GPI anchored proteins) only have a transient or minor role in macrophage invasion, 2) that there is considerable functional redundancy in the surface glycocalyx and/or loss of some components can be compensated for by the acquisition of equivalent host glycolipids, 3) the expression of specific nutrient transporters is essential for life in the macrophage and 4) the role(s) of some surface components differ markedly in different Leishmania species. These mutants will be useful for identifying other surface or intracellular components that are required for virulence in macrophages. PMID- 15357215 TI - Leishmaniasis: current status of vaccine development. AB - Leishmaniasis, a spectrum of diseases caused by various forms of Leishmania has become a major health problem all over the world. Vaccination against leishmaniasis has passed through many developmental stages beginning with the ancient practice of 'leishmanization'. Due to various problems and difficulties associated with traditional vaccines, the interest has been shifted to novel approaches of vaccination like DNA vaccination, vaccination with live vectors encoding leishmanial antigens and finally to designer vaccines. In an effort towards developing an anti-leishmanial vaccine, our laboratory has been working on various genes present in an amplified locus of Leishmania known as the 'LD1 locus'. Two genes, ORFF and BT1 (previously ORFG), are part of the multigenic LD1 locus on chromosome 35. BT1 encodes a biopterin transporter, while the function of ORFF gene product is unknown. Immunization of mice with recombinant ORFF (rORFF) and BT1 proteins, individually, or in combination, conferred partial protection against challenge with Leishmania donovani. We also tested the protective efficacy of ORFF DNA vaccine in BALB/c mice model and found that the level of protection was significantly higher than that of ORFF protein. Protection conferred by ORFF DNA vaccine correlated with significant levels of in vitro splenocyte proliferation and low levels of antigen-specific antibodies. There was a preferential production of IFN-gamma compared to IL-4, which indicated the induction of a protective Th1 response, by the DNA vaccine. Thus, DNA immunization may offer an attractive alternative strategy against leishmaniasis. We present here the current status of vaccine development against leishmaniasis. PMID- 15357216 TI - Macrophage specific drug delivery in experimental leishmaniasis. AB - Macrophage-specific delivery systems are the subject of much interest nowadays, because of the fact that macrophages act as host cells for many parasites and bacteria, which give rise to outbreak of so many deadly diseases(eg. leishmaniasis, tuberculosis etc.) in humans. To combat these deadly diseases initially macrophage specific liposomal delivery system were thought of and tested in vivo against experimental leishmaniasis in hamsters using a series of indigenous or synthetic antileishmanial compounds and the results were critically discussed. In vitro testing was also done against macrophages infected with Leishmania donovani, the causative agent for visceral leishmaniasis. The common problem of liposome therapy being their larger size, stability and storage, non ionic surfactant vesicles, niosomes were prepared, for their different drug distribution and release characteristics compared to liposomes. When tested in vivo, the retention capacity of niosomes was found to be higher than that of liposomes due to the absence of lipid molecules and their smaller size. Thus the therapeutic efficacy of certain antileishmanial compounds was found to be better than that in the liposomal form. The niosomes, being cheaper, less toxic, biodegradable and non-immunogenic, were considered for sometime as suitable alternatives to liposomes as drug carriers. Besides the advent of other classical drugs carriers(e.g. neoglycoproteins), the biggest challenge came from polymeric delivery vehicles, specially the polymeric nanoparticles which were made of cost effective biodegradable polymers and different natural polymers. Because of very small size and highly stable nature, use of nanoparticles as effective drug carriers has been explored in experimental leishmaniasis using a series of antileishmanial compounds, both of indigenous and synthetic origin. The feasibility of application in vivo, when tested for biological as well as for other physicochemical parameters, the polymeric nanoparticles have turned out to be the best and thus may be projected for effective use in the clinics. PMID- 15357217 TI - Role of chemokines in Leishmania infection. AB - Chemokines are a growing group of chemoattractant cytokines that play important roles in physiological as well as pathological processes. Their roles in various aspects of pathogenesis and inflammation have come to light in the past decade or so. It is becoming increasingly clear that chemokines play a major, perhaps decisive role in Leishmania infections. In this review, we recapitulate important works linking the chemokine system with relation to visceral and cutaneous leishmaniasis over the past decade and attemptto put it all together to propose a single yet unfinished model to account for all the findings. PMID- 15357218 TI - Progress in vaccine research and possible effector mechanisms in visceral leishmaniasis. AB - Visceral leishmaniasis represents a serious public health concern in endemic regions and is rapidly emerging as an opportunistic infection in HIV patients. The disease is difficult to diagnose and prevent, and available treatment is associated with toxicity and drug resistance. Even though significant headway has been made in the development of vaccines against cutaneous leishmaniasis, visceral leishmaniasis has received limited attention. The fact that a large proportion of the people living in endemic areas have self-resolving subclinical infection and individuals once recovered are immune to reinfection provides a rationale for designing immunoprophylactic strategies against visceral leishmaniasis. The primary aim of this paper is to review advances in vaccination strategies against visceral leishmaniasis, suggesting possible effector mechanism leading to resistance. It also covers the role of immunostimulators and gives an account of the adjuvants used against visceral leishmaniasis. Vaccine strategies in different established experimental models have also been dealt with which can provide potential leads for their application in humans. In light of the available observations made during the course of studies performed on experimental models of visceral leishmaniasis there is increasing evidence that a successful approach towards a vaccine involves the requirement of Th1 subset of CD4+ cells along with Th2, CD8+, and B cells. In this review we present the possible mechanism of interaction of these cells and their effector molecules in providing resistance against visceral leishmaniasis for the future design of effective vaccine against this disease. PMID- 15357220 TI - Junkie love. PMID- 15357219 TI - DNA topoisomerases in life and death: implications in kinetoplastid protozoa. AB - Current biomedical research has its focus on the search for newer intervention strategies to control public health impact of parasitic diseases. The dramatic advances of molecular and cellular biology in recent times have provided opportunities for discovering and evaluating molecular targets for drug designing, which now form a rational basis for the development of improved anti parasitic therapy. DNA topoisomerases, the "cellular magicians" involved in nearly all biological processes governing DNA, have emerged as one such biological target. Over the last two decades, interest in topoisomerases has expanded beyond the realm of the basic science laboratory into the clinical arena. This review aims at providing a comprehensive insight into the biology of DNA topoisomerases and also focus on its evolution as a drug target in the unicellular kinetoplastids. PMID- 15357221 TI - Parents experiences of their child's care during hospitalization. AB - Driven by the institutionalization of healthcare, and the attendant financial and regulatory issues, health caregivers now need to objectively define and assess the quality of care that is delivered to patients. Measures of quality care for children lag behind the measures that are utilized for adults. To date, little is understood about how quality care for children is identified and measured. Pediatric literature acknowledges, but has poorly studied, the relationship between health care providers and parents in providing and evaluating quality care for children. This paper applies qualitative descriptive theory that is informed by grounded theory to explore parents' experiences of the care they received for their hospitalized children. From a convenience sample of six parent participants, this study finds that parents experience childcare in a hospitalized setting in terms of four interconnecting, circular processes: (a) facing boundaries, (b) attempting to understand, (c) coping with uncertainty and (d) seeking reassurance from caregivers. The experiences shared by the parent participants in this study convey new meaning to the interactional nature of the parent-caregiver relationship. In particular, the researcher finds that parents use the parent-caregiver relationship to help cope with their child's condition, and this in turn influences the parents' sense that their and their child's needs are being met. PMID- 15357223 TI - Managing caregiving at home: Thai caregivers living in the United States. AB - The researchers designed a qualitative descriptive study informed by grounded theory and ethnography to generate a beginning substantive theory that illuminates the process of caregiving in Thai families living in the United States. A purposive sample of seven Thai caregivers of chronically ill relatives in the western and the midwestern regions of the U.S. participated in interviews, provided observations and wrote memoranda. Three categories emerged from "managing caregiving at home:" (a) being caregivers, (b) the consequences of caregiving and (c) coping with the difficulties of caregiving. The study participants describe caregiving as a willing burden and an unavoidable duty, attitudes which are clearly influenced by Buddhist beliefs. The results of this study provide nurses with a better understanding of the caregiving process, especially as it exists in the Thai American population. PMID- 15357222 TI - From mothers to daughters: transgenerational food and diet communication in an underserved group. AB - Studies show that young girls are highly influenced by their mothers' attitudes toward food and body image. To investigate the nature of the information about food that mothers transfer to their daughters, the researchers used focus groups comprising African American women. Results suggest that when educators work with low-income African American women, they should be cognizant of overeating from the perspectives of early food scarcity, reverence for the mother's authoritative role in the family and respect for cultural differences in body size norms. In addition, nutrition educators should counsel young mothers to understand the impact of their eating habits on their daughters. PMID- 15357224 TI - Evidence for racial disparity in cardiac transplantation survival rates. AB - Studies show that survival rates of cardiac transplantation are lower for African American transplant recipients than for Caucasians. Due to similarities in terms of etiology, treatment modalities and responses to these modalities, the authors examine some potential variables for survival of cardiac transplantation that have been well documented in both the renal transplantation literature and the medically managed congestive heart failure literature. The authors also discuss access to care, socioeconomic factors and immunological differences in attempting to identify relevant factors for survival of cardiac transplantation. PMID- 15357225 TI - Health disparities in practice and research for aging women with cancer. AB - The purpose of this article is to examine health needs as well as disparities in practice and research pertaining to aging women with cancer. Given the increasing number of aging women with cancer and the concomitant challenges in survivorship, caregivers need research and improved clinical care to identify needs and enhance cancer survivors' quality of life. The authors use inner strength as a theoretical framework to discuss examples of practice and strategies to improve health outcomes and assess health needs in aging women with cancer. Because researchers find it difficult to identify gender-specific and culturally appropriate instruments, the authors conducted a pilot test on an inner strength questionnaire to identify potential problems in instrumentation and data collection with aging women. Ultimately, a therapeutic partnership between patient and care provider should be based on mutual goal setting and identifying all the dimensions of a women's health status with valid and reliable research instruments and practice strategies. PMID- 15357226 TI - The staffing shortage's far-reaching effects. PMID- 15357227 TI - Managing diversity: the differing perceptions of managers, line workers, and patients. AB - This study assesses how hospital managers, line workers, and patients understand the impact of patient and workforce diversity on the delivery of hospital care. Data come from managerial interviews and staff and patient focus groups at five voluntary hospitals in New York City. Three findings emerge from the research. Cultural competence is critical to the provision of quality care, but the perceived significance of cultural competence depends on one's role in the organization. Language remains a problem for patients and staff, although all of the hospitals have extensive translation services. Compared with management, staff and patients identify diversity to be more of a problem. Differing perceptions about the meaning of diversity must ultimately be reconciled to effectively provide care. PMID- 15357228 TI - Embracing conflict: building a healthy community. AB - All human dynamics include the potential for conflict. Communication processes have deeply embedded in them all the elements of essential conflict. The acknowledgment of differences across the human community is a recognition that all conflict is normative. In healthcare systems, leaders must recognize this factor as an essential part of the expression of the leadership role. Therefore, understanding conflict, applying conflict resolution strategies in the leader's role, building approaches to addressing essential conflict, and resolving it are critical to effective leadership. Understanding the elements of conflict, the processes associated with managing conflict, and the characteristics of conflict resolution are outlined here as essential to the exercise of the leadership role at every level of the organization. PMID- 15357229 TI - Different integration processes of patient satisfaction among four groups. AB - We examined integration processes of patient satisfaction among four groups of patients and found that these groups of patients combined their health care attribute reactions differently to form their overall satisfaction. For the study, we used an emerging noncompensatory model in health care and considered an interaction effect in the analysis. We discuss the implication of the different integration processes of patient satisfaction for health care managers and make practical suggestions for more effective and efficient means of increasing patient satisfaction. PMID- 15357230 TI - Collaboration to improve services for the uninsured: exploring the concept of health navigators as interorganizational integrators. AB - This study examines one specific type of collaboration activity in a community health collaborative. The focus of the research is the operations level where services are coordinated, patients are referred, and access may be improved for vulnerable populations. Using a case study approach, we examine the creation of a new interorganizational role, the "health navigator" in the Miami-Dade Community Access Program. Drawing on traditional theoretical perspectives on coordination, we present the health navigator as an "integrator" among a complex set of safety net providers and public health insurance programs. PMID- 15357231 TI - Opening the black box: measuring hospital information technology capability. AB - Recently, health care investment in information technology (IT) has experienced a significant increase. Paralleling this increase has been an increase in IT capabilities. Despite the interest in and promises of IT in the health care setting, there is a paucity of empirical research that has attempted to define an organizational measure of IT capability. The dearth of research has contributed to the traditional belief that IT is perceived as a "black box," whereby organizational resources enter the box as "inputs" and are somehow transformed into positive outcomes for an organization. However, for positive outcomes to be realized, these outcomes must be measurable. This research uses a stakeholder perspective to develop a theoretically specified measure of IT capability. A latent construct, IT munificence, is proposed using tenets from diffusion of innovation theory and strategic contingency theory. The construct is tested using a sample of 1,545 acute care hospitals located in the United States. IT munificence fits the study data well, supporting the hypothesis that IT munificence represents a strategy of hospital IT capability. PMID- 15357232 TI - Strategy development in small hospitals: stakeholder management in constrained circumstances. AB - Small hospitals form a vital part of the health care safety net, serving communities that would often otherwise lack acute care. It is, therefore, important to understand how strategies unfold in these organizations. We used semistructured interviews to ask chief executive officers (CEOs) of seven small hospitals in Pennsylvania how they viewed their competitive environments and how their strategies evolved. Systematic semi-inductive analyses of these data reveal two major themes. First, CEOs of small hospitals perceive highly dynamic and hostile environments but do not stress complexity. Second, continual negotiations with key stakeholders facilitate the translation of CEOs' insights into organizational strategies. PMID- 15357233 TI - The supply of uncompensated care in Pennsylvania hospitals: motives and financial consequences. AB - The need for uncompensated care has increased during a period in which hospitals are confronted with public and private-sector fiscal pressures. Using a panel design (1995--1998) on Pennsylvania private, not-for-profit general hospitals, we found the provision of uncompensated care is positively associated with financial surpluses, the provision of uncompensated care by neighboring hospitals, bed capacity, proportion of outpatient visits that are emergency, and the unemployment rate (a proxy for need for uncompensated care). Other analysis found that the provision of uncompensated care was not associated with operating surplus, except in hospitals that provide very large amounts of uncompensated care. Provision of services to Medicaid patients and HMO penetration had a negative impact on profitability. PMID- 15357234 TI - The influence of time on changes in health status and patient satisfaction. AB - This article develops a framework that investigates the impact that a change in health status has on satisfaction with structure and process elements of health care delivery over time. We develop and present a structural equation model that encompasses these relationships based on a survey of 284 consumers of health care services. The results indicate that health status is directly related to satisfaction with process of care elements but not directly related to structural elements. These results contribute to the literature by examining the impact of health status on satisfaction in two distinct elements related to structure and process. PMID- 15357235 TI - Care access in rural areas: what leverage mechanisms do regulatory agencies have in a public system? AB - Based on observations of the government's vital role in enhancing rural health care accessibility, in this article, we analyze solutions implemented by Quebec's public health system by highlighting the limitations in incentives used to remedy the dearth of rural medical resources and the consequent interest in ameliorating health care accessibility through service integration. The current challenge lies in fostering cooperation between health care institutions not subject to market incentives and integrating private practice physicians into the public system. To this end, regulatory agencies in public systems use four main leverage mechanisms: formal power, economic power, influence, and commitment, as illustrated in an experiment conducted in rural Quebec. PMID- 15357236 TI - Caring behaviors among nurses: fostering a conversation of gestures. AB - Caring behaviors have been shown to financially benefit business in general and the health care industry in particular. Nursing presence is a concept representative of caring behaviors within the nursing profession. This article suggests a means through which nursing presence might be identified and taught. Patients may, as a result. increasingly view their assigned nurses as caring in attitude and behavior. PMID- 15357237 TI - Environmentally-friendly building strategies slowly make their way into medical facilities. PMID- 15357238 TI - [Structure and organization of the activities of the pathology services for adults in St. Petersburg]. AB - Principles of organization of St. Petersburg pathology service are outlined. The service covers the Consultative and Methodological Council, the Department of Management and 53 departments including 12 departments attached to the city pathology bureau. The main problems of the service concern inadequate financing and material-technical support, lack of specialists in conditions of increasing amount of work (25,870 autopsies, about a million of intraoperative biopsy examinations, 72,000 cytological examinations performed in 2001). PMID- 15357239 TI - [Contribution of the pathology chair of the Military Medical Academy to the development of military pathological anatomy]. AB - In 1930-1940 the pathology chair of the Military Medical Academy participated in organization of military pathological anatomy service. The activity of the chair staff goes in two directions: management of pathology service activity in the army both at peace and war time and research in combat pathology. PMID- 15357240 TI - [Anichkov's views on the role of macrophages in atherogenesis and evolution of such views up to now]. AB - The appearance of macrophages in arterial wall in early morphogenesis of atherosclerosis was noted by N. N. Anichkov in 1940. The macrophages originate partly from monocytic hematogenous elements, partly from subendothelial cells. The authors show that macrophages of monocytic origin play a leading role in atherogenesis-related complex cellular-molecular alterations in the zone of arterial lesions. A cascade of cellular reactions with participation of macrophages determines a chronic course of the process by type of immune inflammation reactions with autonomic regulation. PMID- 15357241 TI - [Thyroid diseases in residents of St. Petersburg]. AB - The degree of tensity of ecological and hygienic state of basic factors can be represented in terms of grades. In St. Petersburg three basic factors reach 5 grades in 7-grade scale. These are radiation, water and atmosphere. By radiation contamination St. Petersburg is on the 5th place after Bryansk, Chelyabinsk, Rostov and Arkhangelsk. As to the number of thyroid diseases, two rises were observed: in the mid 80s and in the mid 90s. A rise in the incidence of thyroid cancer was particularly evident in 1991-1995. The dominating form was papillary carcinoma. PMID- 15357242 TI - [Ultrastructural changes in the cortex of cerebral hemispheres in acute and subacute intoxication with natural gas containing hydrogen sulfide]. AB - Grave destructive changes of myelin nerve fibers in the cortex of the cerebral hemispheres were found in rats intoxicated with hydrogen sulfide gas of the Astrakhan gas deposit. Repeated exposures to natural hydrogen sulfide containing gas in subacute chronic intoxication (100 mg/m3 by H2S) damage predominantly myelin areas corresponding to myelin incisions which are most sensitive to this gas. PMID- 15357243 TI - [Pathomorphology of meningitis caused by Haemophilus infection in children]. AB - Pathomorphological study of the brain of 14 children who died of Hemophilus bacillus infection shows that this disease is characterized by damage to microvessels with formation of microthrombi followed by development of necrosis foci in the brain matter. The maximal damage of the vessels was observed during two first days of the disease in the pia mater and foci of microinfarction were located in the subpial cortex zone only. With the disease progress microvessels of the brain cortex were affected and foci of damage blended together and were found in both cortex and white matter. PMID- 15357244 TI - [Morphology of oculomotor muscles and their nervous apparatus in lateral amyotrophic sclerosis in conditions during long-term artificial lung ventilation]. AB - Light and electron microscopy, histochemical study of acetylcholinesterase (ACE) were used in examination of the state and innervation of ocular muscles in autopsy material from a patient who died of amyotrophic sclerosis (ATS). The patient had lived 14 years under artificial lung ventilation, ATS was diagnosed 22 years before the death. Light microscopy demonstrated the intactness of the muscle fibers and the presence of three types characteristic of the extraocular muscles: thin, granular and rough. Besides typical differences structural changes were observed in some muscle fibers of the myopathic character and inclusions not limited by membrane having filiform or granular structure. Ocular muscles had an intensive innervation. Nervous fiber terminals revealed by a reaction for ACE were represented by single motor plaques and multiple cluster-like endings. Ultrastructurally, nervous endings of two types differed by terminals and fold expression on the postsynaptic membrane. There were no pathological changes in axons and myelin. Thus, ocular muscles were not affected as well as their nervous apparatus at completion of AMS, this indicating the noninvolvement of this muscular allotype in a specific degenerative process. PMID- 15357245 TI - [Phenotypes of smooth muscle cells in carotid arteries in Takayasu's disease]. AB - The phenotype of smooth-muscle cells (SMC) of carotid arteries in health and Takayasu's disease were studied using immunofluorescence. In contrast to unaffected carotid artery, medial SMC were represented by heterogenic cell population. Many SMC in this disease expressed fibronectin with ED-A sequence, contained cytokeratin 8 and were deprived of h-caldesmon. Similar phenotypic characteristics are typical for SMC of embryonal phenotype. SMC with this phenotype are observed in the thickened intima of the carotid arteries in Takayasu's disease. It is supposed that SMC precursors in arterial media are activated in this disease, migrate into subendothelial layer and are responsible for the thickening of the intima. PMID- 15357246 TI - Epstein-Barr virus infected lymphoepithelial carcinomas of the salivary gland in the Russia-Asia area: a clinicopathologic study of 160 cases. AB - In order to determine the prevalence of the Epstein-Barr virus (EBV) infection in salivary gland lymphoepithelial carcinomas (LEC), we have collected 160 cases from Asian countries and Russia. All the cases examined by PCR for EBV DNA BamHI fragment and in-situ hybridization for EBER-1, EBV encoded small RNA, showed positivity for EBV infection in LEC cells, while no positive signals were found in any other salivary neoplasm examined. The incidence of LEC was highest in Guanzhou, followed by Shanghai and Chengdu and lowest in the northern parts of China, Seoul, Niigata, and Moscow. The mean age of the patients with LEC was 43.9 years with no sex predilection. The Chinese patients were of the Han race, only including minor races. There were ninety-five cases found with LEC in the parotid gland (75%), 20 in the submandibular gland (5%), and 28 in the minor salivary gland (20%). Histologically, the LECs were classified into two types: small nest type and large nest type. The latter type consisted of large-sized tumor cell nests and dense lymphocytic stromata and more frequently occurred in the minor salivary gland. The former consisted of small-sized tumor cell nests with fibrous and lymphocyte-depleted stromata, which were more frequently found in the parotid gland. The results indicated that EBV infection and certain geographic factors play important roles in the pathogenesis of the salivary LEC. PMID- 15357247 TI - Gonad development in Drash and Frasier syndromes depends on WT1 mutations. AB - The study of the gonads of 8 cases of Drash syndrome (6 ambiguous males, 2 females) and of 2 Frasier syndrome shows that WT1 mutations gives a dysgenetic testis which is the cause of the genital ambiguity observed at birth. By contrast the same mutations have no effect on ovary development giving normal females. However intron mutations in KTS with isoforms imbalance of WT1 proteins cause streak gonads with a female phenotype in XY patients. In consequence WT1 mutations are the cause of a spectrum of male genital malformations associated with glomerulonephritis and tumors. The absence of WT1 protein detection in sertoli cells shown by immunohistochemistry for 3 cases suggests an imprinting effect of the normal WT1 allele promotor rather than a low level of protein production. A caryotype is mandatory for a correct diagnosis. PMID- 15357248 TI - Molecular pathology of papillary, follicular and Hurthle cell carcinomas of the thyroid. AB - In the review some of the molecular alterations that can relate to the neoplastic development of well differentiated thyroid carcinomas (follicular and papillary carcinoma) and of the so-called Hurthle cell carcinomas are described. Diagnostic and/or prognostic significance of the most recent findings is discussed. PMID- 15357249 TI - [Mycotic lesions in the brain in chronic opium-ephedrone addiction]. AB - Histological examination of the brain of a opium-ephedrone addict has detected large spherical cells resembling Coccidioides or Paracoccidioides spherules. Roundish and oval structures were found in subependymal space of the fourth ventricle. These structures may be spores of many fungi. Areas of normal structure and these with productive inflammation were observed in the surrounding nervous tissue. PMID- 15357250 TI - [Kidney carcinosarcoma]. AB - A kidney tumour in a 36 year old female is described. The tumor was soft, of gray brown color, size 12 x 11 cm. Histological, immunohistochemical and ultrastructural findings have demonstrated the signs of biphasic epithelial and mesenchymal cell differentiation. These data allowed the authors to define this tumor as a renal carcinosarcoma. Histogenesis of this tumor and differential diagnosis with other malignant renal tumors are discussed. PMID- 15357251 TI - [Morphology of the inferior vena cava in the area of an antiembolic cava-filter implanted many years ago]. AB - A case of a 85 year old male who died of uremia is described. An antiembolic cava filter (AECF) implanted probably in the 1960s was found at the level of the kidney veins. Macroscopically, some parts of AECF penetrated through the wall of vena cava. Histologically, fibrous rods were thicker than the wall of the vein, were covered with endothelium and made the lumen of the vein narrow. Recanalization of the fibrous areas was observed. Filters of a new construction are deprived of negative aspects of the old ones. PMID- 15357252 TI - [Methods of silver impregnation in morphological diagnosis of gastric syphilis]. AB - Comparative assessment of methods of silver impregnation for pathohistological identification of Treponema pallidum in gastric syphilis was performed. Levaditi impregnation is preferable in cases of gastrectomy. Silver impregnation in histological slides is performed for studying endoscopic biopsies in view of their small size as well as for retrospective analysis when paraffin blocks only are available. Steiner's method in modification with the use of microwave stove allows better identification of Treponema pallidum than impregnation by Vartin Starri. Helicobacter pylori may also be identified by silver impregnation. Association of syphilitic and Helicobacter infections deteriorates inflammatory and destructive lesions in the stomach. PMID- 15357253 TI - [In memory of professor A. V. Rusakov]. PMID- 15357254 TI - [The 4th all-Russian workshop on immunohistochemical diagnosis of human tumors]. PMID- 15357255 TI - [Current status of tobacco smoking in Japan]. PMID- 15357256 TI - [The Japanese Respiratory Society promotes a smoke-free society]. PMID- 15357257 TI - [Passive smoking]. PMID- 15357258 TI - [Smoking attitude among hospital workers]. PMID- 15357259 TI - [Smoking cessation education]. PMID- 15357260 TI - [Stop smoking program]. PMID- 15357261 TI - [Countermeasure against passive smoking in workplaces]. PMID- 15357262 TI - [Electronic clinical pathway for community acquired pneumonia (e-CP CAP)]. AB - Clinical pathway is a useful strategy for the management of risk and cost for medical care. We developed an electronic clinical pathway (e-CP) for community acquired pneumonia (CAP). We programmed all medical procedures, including physical and laboratory examinations, medical care, prescriptions, diets, limitations of activities, and patient education on a computer. With e-CP, we achieved 30% cost reduction (34048 points vs. 24338 points), 25% shortening of admission duration (10.77 days vs. 8.03 days) and 21% shortening of the duration of intravenous antibiotics administration (8.22 days vs. 6.47 days). Introduction of computer-aided clinical pathway for CAP may contribute to the standardization of medical care for patients with CAP. PMID- 15357263 TI - [A case of primary pleural leiomyoma]. AB - The chest radiograph = of a 33-year-old man showed a mass shadow in the right hilum. CT scanning film revealed a 3 x 2 cm tumor lying against the right pulmonary artery. Bronchoscopy and pulmonary arteriography showed almost normal findings. VATS tumor extirpation was performed. Macroscopically, the tumor was located in a major fissure. The resected specimen was compatible with leiomyoma. Because no organ of origin of the leiomyoma, other than the pleura, was detected, this case was diagnosed as a primary pleural leiomyoma. Primary pleural leiomyoma is very rare, and this report presents the first such case reported in Japan. PMID- 15357264 TI - [A case of chronic pulmonary paracoccidioidomycosis]. AB - In a 43-year-old Japanese Brazilian who came to Japan in 2001, since subjective symptoms such as cough, sputum, and dyspnea on exertion had become severe, he was referred to our hospital because of suspicion of pulmonary tuberculosis in chest radiography and CT findings. A chest radiograph of initial examination showed interstitial shadows in both lungs with nodular, infiltrative or cavitary changes. No Mycobacterium tuberculosis was found. The mycetocyte with multipolar budding resembling the steerage of a ship, which was characteristic of Paracoccidioides was observed in sputum and transbronchial lung biopsy specimens. We cultured a fungus to show dimorphism of temperature dependency, and a diagnosis of chronic lung paracoccidioidomycosis was arrived at. By administration of ITCZ 200 mg/day, the chest radiography findings and clinical manifestations were improved. This case seems to be worthy of reporting in Japan since the affected site or organ was limited to the two lungs with multiple cavitary lesions and fibrotic changes on radiographic examination, and final diagnosis was made by cytology of sputum and pathology of TBLB specimens. PMID- 15357265 TI - [Thymic carcinoma. A clinicopathological study of seven patients]. AB - The treatment of seven cases of thymic carcinoma is reported. According to the Masaoka classification of the disease, there were five cases of stage III and two of stage IVb. Five were identified histopathologically as squamous cell carcinoma, one as undifferentiated carcinoma, and one as small cell neuroendocrine carcinoma. Surgery was performed during the course of chemotherapy and radiotherapy in 5 cases, and in 2, the organs infiltrated by neoplastic cells were partially excised together. Radiotherapy was performed as adjuvant therapy in one case of partial excision. In another case, after six years of chemotherapy and radiotherapy that yielded a partial response (PR), cancerous infiltration of the chest wall occurred, and partial removal of the chest wall became necessary. After the remaining case showed a PR to chemotherapy, complete macroscopic excision of the tumor was undertaken, but mediastinal lymph gland metastasis was present, and so radiotherapy was also initiated. In two cases, surgery was not performed. One of these was treated with chemotherapy alone, the other, only with radiotherapy. The mean survival period of the five patients who died as a result of their tumors was 44.8 months. One of the two who survived for over four years had squamous cell carcinoma, the other, undifferentiated carcinoma. The case of small cell neuroendocrine carcinoma was assessed as having a PR, and so complete extraction of the thymic neoplasm was carried out, and followed with radiotherapy. This patient is still alive. PMID- 15357266 TI - [A case of acute eosinophilic pneumonia with a disassociation between the KL-6 level and SP-A and SP-D levels in the serum and BAL fluid]. AB - A 45-year-old Japanese electrical engineer was admitted to our department of internal medicine on August 12, 2003, because of a sudden high fever and severe hypoxic respiratory failure. At a barbecue with his family on August 3 beside a nearby river, he had been exposed to the smoke. From August 4 to 11, he had suffered fatigue, fever, dry cough and progressive dyspnea. On admission, his SpO2 was 84%, and computed tomography scanning showed patchy ground glass opacity, thickened bronchial walls, and bilateral pleural effusions. The eosinophil count in the bronchoalveolar lavage fluid (BALF) was increased to 52.4%. Noticeably, the KL-6, SP-A and SP-D levels in the serum were elevated to 197 U/ml, 188 ng/ml and 137 ng/ml, and their levels in BALF had also increased to 225 U/ml, 890 ng/ml and 1110 ng/ml, respectively. The lymphocyte stimulation test was negative, and the cultures of blood and BALF did not grow any pathogens. The patient had smoked 1 pack of per cigarettes day for 25 years and showed no sign of atopic illness. Acute eosinophilic pneumonia (AEP) was diagnosed, and responded dramatically to treatment with oxygen and corticosteroids. The dissociation between the normal KL-6 levels and the elevated SP-A and SP-D levels in the serum and BAL fluid may play an important role in cases of AEP. PMID- 15357267 TI - [Two cases of acute respiratory failure associated with pneumonia requiring mechanical ventilation before diagnosis of amyotrophic lateral sclerosis]. AB - We present two cases of acute respiratory failure requiring mechanical ventilation before diagnosis of amyotrophic lateral sclerosis (ALS). The patients were men of 60 (patient 1) and 74-years old (patient 2), both of whom exhibited acute respiratory failure requiring mechanical ventilation. Diagnoses of ALS were made because of continuous aspiration caused by bulbar palsy in patient 1, and, in patient 2, because of the progressive muscle atrophy that occurred during unsuccessful attempts to wean the patient from ventilatory support. Physicians should be aware of the possibility of ALS in cases of acute respiratory failure, CO2 narcosis, continuous aspiration, and difficulty of weaning from mechanical ventilation. PMID- 15357268 TI - [A case of successful desensitization therapy for isoniazid-induced pneumonitis]. AB - A 65-year-old man was admitted to hospital for treatment of pulmonary tuberculosis. He was treated with isoniazid (INH), rifampicin (RFP), ethambutol (EB), and pyrazinamide (PZA). On the 14th day, he developed a fever and interstitial pneumonia, which improved promptly after discontinuation of the antituberculous drugs. Drug lymphocyte stimulation tests against INH, RFP and PZA were negative. However, the provocation test on INH (only) was positive, leading to a diagnosis of pneumonitis caused by INH. We then tried desensitization of INH over a period of two weeks, which was successful and occurred without any clinical event. In the past, five cases of INH-induced pneumonitis were reported, but desensitization of INH did not occur in any. We conclude that physicians should be aware not only of paradoxical reactions but also of drug-induced pneumonitis when a new pulmonary infiltrate develops in the course of tuberculosis treatment. Furthermore, drug desensitization may be possible in some cases of drug-induced pneumonitis. PMID- 15357269 TI - [A case of Crohn's disease with broncho-bronchiolitis]. AB - A 41-year old man was hospitalized for abdominal pain and melena. Two years before, he had contracted Crohn's disease, which was brought into remission by using mesalazine and oral corticosteroids. When Crohn's disease appeared to flare up again, the dose of corticosteroids was increased. Afterwards, when a gradual decrease in this dose was initiated, melena developed once again, along with a dry cough. Chest radiography revealed diffuse patchy shadows. A significant increase of lymphocytes was observed in the bronchoalveolar lavage fluid. Mesalazine was suspended and steroid pulse therapy was initiated in combination with azathioprine. Although the symptoms were alleviated and the patchy shadows disappeared, the bronchiectatic shadows remained. A broncho-bronchiolitis associated with Crohn's disease was finally diagnosed despite the need for a differential diagnosis of mesalazine-induced disease due to the long-term administration of mesalazine before the onset of the pulmonary lesion. Also, the pathological findings were inconsistent with many other reported cases, the onset of the pulmonary lesion coincided with the flare-up of an intestinal disease, and the pulmonary shadows remained long after the suspension of mesalazine. PMID- 15357270 TI - [Six cases of thymic carcinoma: a clinical review]. AB - Thymic carcinoma is rarer than thymoma and carries a very poor prognosis. No standard treatment has yet been established. Chemotherapy and radiation therapy are usually given to patients in whom surgery is not indicated. We clinically review six patients with thymic carcinoma (three men and three women) who were admitted to our hospital from 1992 through 2002. Their ages ranged from 56 to 81 years (median 70.5). Four patients had chest pain, two were unable to swallow, and in one, who was asymptomatic, the tumor was detected by chance. Histologically, five cases were squamous cell carcinomas, and one was a lymphoepithelioma-like carcinoma. Four of the six patients had distant metastasis at diagnosis. Only one patient was able to undergo surgery, two received radiation therapy alone, and three were administered chemoradiation. The response to the chemoradiation was progression of the disease in two patients and no change in one. The response to radiation therapy was partial response in one patient and no change in one. In summary, chest pain is the most common symptom of thymic carcinoma. A symptomatic case suggests a poor prognosis resulting from the clinical behavior of thymic carcinoma, such as early distant metastasis or direct invasion of surrounding organs. Histologically, squamous cell carcinoma is the most common, and thymic carcinoma appears to be generally resistant to chemotherapy and radiotherapy. PMID- 15357271 TI - [A case of organizing pneumonia associated with myelodysplastic syndrome (MDS)]. AB - A 53-year-old man was referred to our hospital because of atypical cells in the peripheral blood. Myelodysplastic syndrome (MDS) was diagnosed by bone marrow examination. Since he had a continuous dry cough, high fever, and air space consolidation in both lower lung fields, he was admitted to our hospital for further examination. Infectious disease, autoimmune disease, vasculitis and neoplasms were excluded. A transbronchial lung biopsy (TBLB) was performed at the right S(10), and histopathological examination of the specimen thus obtained suggested an organizing pneumonia (OP) associated with MDS. Oral prednisolone 30 mg was administered daily, and the symptoms and radiographic findings immediately improved. Accurate diagnosis of the pulmonary involvement associated with patients with MDS is sometimes difficult to make. However, either BOOP or OP should be considered as a differential diagnosis in case the patient presents with continuous coughing, high fever, and air space consolidation. Interventional examinations such as TBLB should be conducted to make a histological diagnosis if the patient's condition allows it. PMID- 15357272 TI - [A case of pulmonary embolic metastasis of choriocarcinoma presenting with multiple pulmonary infiltrative shadows]. AB - A 34-year-old woman visited our hospital complaining of dry cough. Chest radiography and computed tomography showed bilateral multiple infiltrative shadows over the lung field. After an initial diagnosis of pneumonia, antibiotics were administered, but the therapy failed to improve the symptoms and abnormalities observed on the chest radiograph. The patient was then admitted to our hospital. The bronchoalveolar lavage fluid (BALF) was slightly bloody, but we were not able to make any specific findings in BALF. In order to confirm the pathological diagnosis, video-assisted thoracoscopic lung biopsy was performed aiming at the right middle and lower lobes. There were bleeding pulmonary infarctions in a biopsy specimen from the right middle lobe. Atypical cells positive for human chorionic gonadotropin (hCG) proliferated in the pulmonary arteries, and so a diagnosis of pulmonary embolic metastasis of choriocarcinoma was made. After the diagnosis, it became clear that urine and serum hCG values were very high. The patient has since received systemic chemotherapy in the gynecology unit at our hospital. Pulmonary embolic metastasis of choriocarcinoma diagnosed by video-assisted thoracoscopic lung biopsy has never been reported in the literature. However, early hCG measurement may have detected this syndrome in the earlier stages, and pulmonary metastasis of choriocarcinoma should be considered in the differential diagnosis of women with past pregnancy presenting with intractable multiple pulmonary shadows. PMID- 15357273 TI - [Hypersensitivity pneumonitis induced by Aspergillus niger--a case report]. AB - A 52-year-old woman was hospitalized because of severe cough in August 1994. She had engaged in culturing roses in greenhouses since 1968, and had developed a cough during the summer of 1990. Chest radiography showed diffuse ground-glass opacity in both lung fields, and she suffered from hypoxemia (PaO2 = 45.6 torr) while breathing room air. The lymphocyte count in the bronchoalveolar lavage fluid was increased, and transbronchial lung biopsy specimens showed lymphocyte alveolitis in the alveolar spaces. After admission, the patient's symptoms improved rapidly without medication. However, on her return to work, the cough and hypoxemia reappeared. In her rose culture, she had used Rockwool, and Aspergillus niger was detected predominantly in the Rockwool. Precipitins against the extracts of Aspergillus niger were detected with the double immunodiffusion test and the inhalation provocation test yielded clinical symptoms. Our diagnosis was hypersensitivity pneumonitis caused by Aspergillus niger. PMID- 15357274 TI - By the numbers. Mounting medical bills. PMID- 15357275 TI - Statins could be the next aspirin. PMID- 15357276 TI - Can you really make fast food healthy? PMID- 15357277 TI - Not for the faint of heart. Boston Scientific's stent recall is a blow to earnings--but an opportunity for investors. PMID- 15357278 TI - Baxter's healthy return. Morgan Stanley's Glenn Reicin says the medical products company's prognosis is finally positive. PMID- 15357279 TI - Healthcare fraud and quality of care: a patient-centered approach. AB - This Article explores the intersection between quality of care and healthcare fraud by examining the extent to which quality-related fraud settlements benefit patients. The author argues that, although the protection of beneficiary health and welfare often is invoked by the federal government as one of the reasons for undertaking anti-fraud efforts, such considerations do not appear to play a large role in many of the settlements that are negotiated. While returning funds to the federal Treasury helps to ensure that the federal healthcare programs remain solvent and continue to serve beneficiaries in the aggregate, it may not adequately address harm to individual patients. Thus, the author concludes it may be time to explore new models of fraud settlements that can provide adequate compensation to the patients who may have suffered harm. PMID- 15357280 TI - The doctor-patient relationship as the business case for quality: doing well by doing right. AB - The idea that healthcare quality in America has lagged behind optimal levels has been a central national policy issue since at least 1998. Reform efforts, however, have failed to acknowledge the critical and unique role physicians play in making quality initiatives real. This Article argues physicians are at the core of any effort to propel quality forward in a significant way and, therefore, must be taken into account directly and without apology. The Article examines the quality-accountability context present in this country. It addresses why the physician nexus on these issues is vital for real change to take place and sets forth a clear statement of what is essential to support the doctor-patient relationship in any quality agenda. Finally, it reiterates the author's previously proposed five principles for quality, as well as her unified field theory of quality, all of which are designed to advance quality in a very different way. PMID- 15357281 TI - Application of the HIPAA privacy rule to employer benefit plans and a compliance theory of statutory interpretation. AB - The application of the federal privacy regulations promulgated pursuantto the Health Insurance Portability and Accountability Act of 1996 (HIPAA) to employer benefit plans is arguably the most conceptually difficult area of a complex law. A purely textual reading of the Rule, when applied to employer plans, results in varying interpretations on some significant issues and puzzling results on others. This Article offers a practical approach for interpreting the rule when clear-cut answers are not provided by the text and DHHS guidance is nonexistent or unclear. In addition, this approach can be applied to the interpretation of other statutes and regulations. PMID- 15357282 TI - Images of health insurance in popular film: the dissolving critique. AB - Several recent films have villainized the health-insurance industry as a central element of their plots. This Article examines three of those films: Critical Care, The Rainmaker, and John Q. It analyzes these films through the context of the consumer backlash against managed care that began in the 1990s and shows how these films reflect the consumer sentiment regarding health-insurance companies and the cost controlling strategies they employ. In addition, the Article identifies three key premises about health insurance in the films that, although exaggerated and incomplete, have significant factual support. Ultimately, the author argues that, despite their passionately critical and liberal tone, these films actually put forward solutions that are highly individualist and conservative, rather than inclusive and systemic. PMID- 15357283 TI - Prompt-pay laws: a state-by-state analysis. AB - The following cases are the result of research performed in all state jurisdictions for any cases addressing "prompt pay." Also included is a state survey of prompt-pay statutes. PMID- 15357284 TI - Denaturing gradient gel electrophoresis analysis of bacterial populations in 5 stage biological nutrient removal process with step feed system for wastewater treatment. AB - Changes in the bacterial populations of a 5-stage biological nutrient removal (BNR) process, with a step feed system for wastewater treatment, were monitored by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S ribosomal DNA fragments. DGGE analysis indicated seasonal community changes were observed, however, community profiles of the total bacteria of each reactor showed only minor differences in the samples obtained from the same season. The number of major bands was higher in the summer samples, and decreased during the winter period, indicating that the microbial community structure became simpler at low temperatures. Since the nitrogen and phosphate removal efficiencies were highly maintained throughout the winter operation period, the bacteria which still remaining in the winter sample can be considered important, playing a key role in the present 5-stage BNR sludge. The prominent DGGE bands were excised, and sequenced to gain insight into the identities of the predominant bacterial populations present, and most were found to not be closely related to previously characterized bacteria. These data suggest the importance of culture-independent methods for the quality control of wastewater treatment. PMID- 15357285 TI - A method for comparing multiple bacterial community structures from 16S rDNA clone library sequences. AB - Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/. PMID- 15357286 TI - Genomic relationship of Salmonella enterica serovar typhimurium DT104 isolates from Korea and the United States. AB - Salmonella enterica serovar Typhimurium DT104 (Salmonella Typhimurium DT104 or DT104) has been emerging as a common pathogen for human in Korea since 1997. In order to compare the genomic relationship and to search for the dominant strains in Korea, we conducted pulsed-field gel electrophoresis (PFGE) and IS200 fingerprinting of 25 epidemiological unrelated isolates from human and animals from Korea and cattle from America. Two Salmonella Typhimurium DT104 isolates from human in Korea and all 8 isolates from American cattle had indistinguishable patterns from the PFGE and IS200 fingerprinting but multidrug-resistant Salmonella Typhimurium isolates, including DT104, from Korean animals had diverse genetic patterns. The data suggest that a dominant DT104 strain might have circulated between Korean and American cattle and that it had a high level of clonality. PMID- 15357287 TI - Characterization of recombinant Drosophila melanogaster myo-inositol-1-phosphate synthase expressed in Escherichia coli. AB - Cloned myo-inositol-1-phpsphate synthase (INOS) of Drosophila melanogaster was expressed in Escherichia coli, and purified using a His-affinity column. The purified INOS required NAD+ for the conversion of glucose-6-phosphate to inositol 1-phosphate. The optimum pH for myo-inositol-1-phosphate synthase is 7.5, and the maximum activity was measured at 40 degrees C. The molecular weight of the native enzyme, as determined by gel filtration, was approximately Mr 271,000 +/- 15,000. A single subunit of approximately Mr 62,000 +/- 5,000 was detected upon SDS polyacrylamide gel electrophoresis. The Michaelis (Km) and dissociation constants for glucose-6-phosphate were 3.5 and 3.7 mM, whereas for the cofactor NAD+ these were 0.42 and 0.4 mM, respectively. PMID- 15357288 TI - Study on persistent infection of Japanese encephalitis virus Beijing-1 strain in serum-free Sf9 cell cultures. AB - Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8 x 10(6) cells/ml in a 500ml spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-1 strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15% of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-1 strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-1 stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-1 strain rose from 1.0 x 10(5) to 1.5 x 10(6) pfu/ml. The infected Sf9 cells could be sub-cultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-1 strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-1 strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied. PMID- 15357289 TI - Schizosaccharomyces pombe rsm1 genetically interacts with spmex67, which is involved in mRNA export. AB - We have previously isolated three synthetic lethal mutants from Schizosaccharomyces pombe in order to identify mutations in the genes that are functionally linked to spmex67 with respect to mRNA export. A novel rsm1 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMexl. The rsm1 gene contains no introns and encodes a 296 amino-acid long protein with the RING finger domain, a C3HC4 in the N-terminal half. The deltarsm1 null mutant is viable, but it showed a slight poly(A)+ RNA accumulation in the nucleus. Also, the combination of deltarsm1 and deltaspmex67 mutations confers synthetic lethality that is accompanied by the severe poly(A)+ RNA export defect. These results suggest that rsm1 is involved in mRNA export from the nucleus. PMID- 15357290 TI - The role of enzymes produced by white-rot fungus Irpex lacteus in the decolorization of the textile industry effluent. AB - The textile industry wastewater has been decolorized efficiently by the white rot fungus, Irpex lacteus, without adding any chemicals. The degree of the decolorization of the dye effluent by shaking or stationary cultures is 59 and 93%, respectively, on the 8th day. The higher level of manganese-dependent peroxidase (MnP) and non-specific peroxidase (NsP) was detected in stationary cultures than in the cultures shaken. Laccase activities were equivalent in both cultures and its level was not affected significantly by the culture duration. Neither lignin peroxidase (LiP) nor Remazol Brilliant Blue R oxidase (RBBR ox) was detected in both cultures. The absorbance of the dye effluent was significantly decreased by the stationary culture filtrate of 7 days in the absence of Mn (II) and veratryl alcohol. In the stationary culture filtrate, three or more additional peroxidase bands were detected by the zymogram analysis. PMID- 15357291 TI - Chlorothalonil-biotransformation by glutathione S-transferase of Escherichia coli. AB - It has recently been reported that one of the most important factors of yeast resistance to the fungicide chlorothalonil is the glutathione contents and the catalytic efficiency of glutathione S-transferase (GST) (Shin et al, 2003). GST is known to catalyze the conjugation of glutathione to a wide variety of xenobiotics, resulting in detoxification. In an attempt to elucidate the relation between chlorothalonil-detoxification and GST, the GST of Escherichia coli was expressed and purified. The drug-hypersensitive E. coli KAM3 cells harboring a plasmid for the overexpression of the GST gene can grow in the presence of chlorothalonil. The purified GST showed chlorothalonil-biotransformation activity in the presence of glutathione. Thus, chlorothalonil is detoxified by the mechanism of glutathione conjugation catalyzed by GST. PMID- 15357292 TI - Effect of titanium-ion on the growth of various bacterial species. AB - There are a number of studies that explain the metabolism and roles of metallic titanium and titaniumion. One of the most intriguing results from these studies is the finding of metallic titanium having no bacteriostatic effects on oral bacterial species. In this research, the effects of titanium-ion on the growth of twenty-two bacterial species, some of which are commonly found in foods such as yoghurt, kimchi, and soy fermented products, were investigated. All but two bacteria, Escherichia coli and Pseudomonas aeruginosa appeared to be sensitive to titanium-ion. These two species were grown on 360 microg/ml of titanium-ions, and they were found to be resistant to the titanium-ion. Both the wild-type and plasmid-cured E. coli showed good growth in a medium with 200 microg/ml of titanium-ions. These results suggest that titanium-resistance was independent from the effects of the plasmid in E. coli. PMID- 15357293 TI - Optimal fermentation conditions for enhanced glutathione production by Saccharomyces cerevisiae FF-8. AB - The influence of feedstock amino acids, salt, carbon and nitrogen sources on glutathione production by Saccharomyces cerevisiae FF-8 was investigated. Glucose, yeast extract, KH2PO4, and L-cysteine were found to be suitable feedstock. Highest glutathione production was obtained after cultivation with shaking for 72 h in a medium containing glucose 3.0% (w/v), yeast extract 3.0%, KH2PO4 0.06% and L-cysteine 0.06%. The glutathione concentration achieved using this medium increased 2.27-fold to 204 mg/l compared to YM basal medium. PMID- 15357294 TI - Transposition of IntAs into the conserved regions of IS3 family elements. AB - Together with the previous reports, my computer survey revealed that several bacteria contain six copies of the type group II intron IntA. The sequence analysis of IntAs showed the high level of homology in the nucleotide sequence (91.9-99.8%). The consensus sequence, 2,270 base pair long, was derived from the nucleotide sequences of all IntA members. The size of the open reading frame intA was 502 amino acids long, that is homologous to reverse transcriptase-like proteins encoded within the group II introns. It was reported that EPEC.IntA and Sf.IntA were inserted into IS911 and IS629, respectively. The sequence of the flanking region IntA was analyzed here. The data show the insertion of EC.IntA into IS629, the insertion of EHEC.IntA into IS3, the insertion of Yp.IntA into IS904-like sequence, and the insertion of EK12.IntA into IS911. Interestingly, these IS elements nested by IntAs were the members of IS3 family elements. The sequences of the IS3 members correspond to the OrfB with the DDE motif conserved in retroviral integrases. Alignment of the flanking sequences of IntAs revealed that the flanking regions -25 to +10 of insertion sites, that are generally believed to be required for the retrohoming, were not strongly conserved. The data presented here suggests that the retrohoming pathway of IntA seems to differ from those of other group II introns. PMID- 15357295 TI - Use of sucrose-agar globule with root exudates for mass production of vesicular arbuscular mycorrhizal fungi. AB - A sucrose-agar globule (SAG) was newly introduced to increase production of the vesicular arbuscular mycorrhizal (VAM) fungal spores, Gigaspora gigantea and Glomus fasciculatum. An SAG inoculum and a sucrose-agar globule with root exudates (SAGE) inoculum were prepared, and their spore productions were compared with a soil inoculum. When the SAGE was used as the inoculum on sucrose-agar medium plates the number of spores was increased (35% more than the soil inoculum). After the soil inoculum and SAGE were inoculated on an experimental plant, Zingiber officinale, the percentage root colonization, number of VAM spores, and dry matter content were analyzed. It was observed that the SAGE showed a higher percentage of root colonization (about 10% more), and increases in the number of spores (about 26%) and dry matter (more than 13%) for the two VAM fungal spores than the soil inoculum. The results of this study suggested that the SAGE inoculum may be useful for the mass production of VAM fungi and also for the large scale production of VAM fungal fertilizer. PMID- 15357296 TI - Formation and dispersion of mycelial pellets of Streptomyces coelicolor A3(2). AB - The pellets from a culture of Streptomyces coelicolor A3(2) that were submerged shaken were disintegrated into numerous hyphal fragments by DNase treatment. The pellets were increasingly dispersed by hyaluronidase treatment, and mycelial fragments were easily detached from the pellets. The submerged mycelium grew by forming complexes with calcium phosphate precipitates or kaolin, a soil particle. Therefore, the pellet formation of Streptomyces coelicolor A3(2) can be considered a biofilm formation, including the participation of adhesive extracellular polymers and the insoluble substrates. PMID- 15357297 TI - Changes in membrane fatty acid composition during entry of Vibrio vulnificus into the viable but nonculturable state. AB - Vibrio vulnificus, a Gram-negative bacterium found in estuarine waters, is responsible for over 95% of all seafood-related deaths in the United States. As a result of a temperature downshift to 5 degrees C, this organism enters the viable but nonculturable (VBNC) state. Changes in the membrane fatty acid (FA) composition of V. vulnificus may be a contributing factor to the ability of this organism to enter into and survive in the VBNC state. This hypothesis was tested by incubating the organism at 5 degrees C in artificial sea water and analyzing the cells' FAs during the initial hours of temperature and nutrient down-shift. Prior to downshift, the predominant FAs were 16:0, 16:1 and 18:0. During the first four hours of downshift, statistically significant changes occurred in 15:0, 16:1, 16:0, 17:0, and 18:0. These results indicate that changes in FA composition occur prior to entry of V. vulnificus into the VBNC state, suggesting that the ability to maintain membrane fluidity may be a factor in this physiological response. Cells in which fatty acid synthesis was inhibited did not survive, indicating that active fatty acid metabolism is essential for entry of cells into the VBNC state. PMID- 15357298 TI - The viable but nonculturable state of Kanagawa positive and negative strains of Vibrio parahaemolyticus. AB - Ingestion of shellfish-associated Vibrio parahaemolyticus is the primary cause of potentially severe gastroenteritis in many countries. However, only Kanagawa phenomenon (hemolysin) positive (KP+) strains of V. parahaemolyticus are isolated from patients, whereas >99% of strains isolated from the environment do not produce this hemolysin (i.e. are KP-). The reasons for these differences are not known. Following a temperature downshift, Vibrio parahaemolyticus enters the viable but nonculturable (VBNC) state wherein cells maintain viability but cannot be cultured on routine microbiological media We speculated that KP+ and KP- strains may respond differently to the temperature and salinity conditions of seawater by entering into this state which might account for the low numbers of culturable KP+ strains isolated from estuarine waters. The response of eleven KP+ and KP- strains of V. parahaemolyticus following exposure to a nutrient and temperature downshift in different salinities, similar to conditions encountered in their environment, was examined. The strains included those from which the KP+ genes had been selectively removed or added. Our results indicated that the ability to produce hemolysin did not affect entrance into the VBNC state. Further, VBNC cells of both biotypes could be restored to the culturable state following an overnight temperature upshift. PMID- 15357299 TI - Molecular investigation of two consecutive nosocomial clusters of Candida tropicalis candiduria using pulsed-field gel electrophoresis. AB - Pulsed-field gel electrophoresis (PFGE) typing was applied to the epidemiological investigation of 21 Candida tropicalis isolates collected from urine specimens of 11 patients and one healthcare worker, in an intensive care unit (ICU) over a 4 month period. Seventeen epidemiologically unrelated strains from 14 patients were also tested to determine the discriminatory power of PFGE. PFGE typing consisted of electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG), using two restriction enzymes (BssHII and SfiI). The EK pattern was the same in all 38 isolates, while REAG using SfiI separated the isolates into nine types. However, 16 different PFGE types were identified by REAG with BssHII, and the same results were obtained when the results of both REAG tests were combined. In serial urinary isolates from 10 patients, all strains from each patient had the same PFGE pattern. While the epidemiologically unrelated strains from 14 patients consisted of 13 different PFGE types, the 20 isolates from the 11 ICU patients fell into only two PFGE types (types C1 and C2), and these apparently originated from the two different outbreaks. All strains of type C1 (n = 12) were isolated from six patients, between November 1999 and January 2000, and all of the type C2 strains (n=8) were isolated from five patients, during January and February 2000. This study shows two consecutive clusters of C. tropicalis candiduria in an ICU, defined by PFGE typing, and also demonstrates that a PFGE typing method using BssHII is perhaps the most useful method for investigating C. tropicalis candiduria. PMID- 15357300 TI - Genetic and phenotypic diversity of (R/S)-mecoprop [2-(2-methyl-4 chlorophenoxy)propionic acid]-degrading bacteria isolated from soils. AB - Twelve mecoprop-degrading bacteria were isolated from soil samples, and their genetic and phenotypic characteristics were investigated. Analysis of 16S rDNA sequences indicated that the isolates were related to members of the genus Sphingomonas. Ten different chromosomal DNA patterns were obtained by polymerase chain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences from the 12 isolates. The isolates were found to be able to utilize the chiral herbicide mecoprop as a sole source of carbon and energy. While seven of the isolates were able to degrade both (R)- and (S)-mecoprop, four isolates exhibited enantioselective degradation of the (S)-type and one isolate could degrade only the (R)-enantiomer. All of the isolates were observed to possess plasmid DNAs. When certain plasmids were removed from isolates MP11, MP15, and MP23, those strains could no longer degrade mecoprop. This compelling result suggests that plasmid DNAs, in this case, conferred the ability to degrade the herbicide. The isolates MP13, MP15, and MP24 were identified as the same strain; however, they exhibited different plasmid profiles. This indicates that these isolates acquired different mecoprop-degradative plasmids in different soils through natural gene transfer. PMID- 15357301 TI - Degradation of phenanthrene by Trametes versicolor and its laccase. AB - Phenanthrene is a three-ring polycyclic aromatic hydrocarbon and commonly found as a pollutant in various environments. Degradation of phenanthrene by white rot fungus Trametes versicolor 951022 and its laccase, isolated in Korea, was investigated. After 36 h of incubation, about 46% and 65% of 100 mg/l of phenanthrene added in shaken and static fungal cultures were removed, respectively. Phenanthrene degradation was maximal at pH 6 and the optimal temperature for phenanthrene removal was 30 degrees C. Although the removal percentage of phenanthrene was highest (76.7%) at 10 mg/l of phenanthrene concentration, the transformation rate was maximal (0.82 mg/h) at 100 mg/L of phenanthrene concentration in the fungal culture. When the purified laccase of T versicolor 951022 reacted with phenanthrene, phenanthrene was not transformed. The addition of redox mediator, 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) or 1-hydroxybenzotriazole (HBT) to the reaction mixture increased oxidation of phenanthrene by laccase about 40% and 30%, respectively. PMID- 15357302 TI - Biochemical quantitation of PM2 phage DNA as a substrate for endonuclease assay. AB - Bacteriophage PM2 has a closed circular form of double stranded DNA as a genome. This DNA from the phage is a useful source for nick-circle endonuclease assay in the fmol range. Due to difficulties in the maintenance of viral infectivity, storage conditions of the phage should be considered for the purification of PM2 DNA. The proper condition for a short-term storage of less than 2 months is to keep the PM2 phage at 4 degrees C; whereas the proper condition for a long-term storage of the PM2 phage for over 2 months is to keep it under liquid nitrogen in 7.5% glycerol. The optimal conditions for a high yield of phage progeny were also considered with the goal to achieve a successful PM2 DNA preparation. A MOI(Multiplicity Of Infection) of 0.03, in which the OD600 of the host bacteria was between 0.3 and 0.5, turned out to be optimal for the mass production of PM2 phage with a burst size of about 214. Considerations of PM2 genome size, and the concentrations and radiospecific activities of purified PM2 DNA, are required to measure the endonuclease activity in the fmol range. This study reports the proper quantitation of radioactivity and the yield of purified DNA based on these conditions. PMID- 15357303 TI - Factors influencing preferential utilization of RNA polymerase containing sigma 38 in stationary-phase gene expression in Escherichia coli. AB - In order to understand the molecular basis of selective expression of stationary phase genes by RNA polymerase containing sigma38 (Esigma38) in Escherichia coli, we examined transcription from the stationary-phase promoters, katEP, bolAP, hdeABP, csgBAP, and mcbP, in vivo and in vitro. Although these promoters are preferentially recognized in vivo by Esigma38, they are transcribed in vitro by both Esigma38 and Esigma70 containing the major exponential sigma, sigma70. In the presence of high concentrations of glutamate salts, however, only Esigma38 was able to efficiently transcribe from these promoters, which supports the concept that the promoter selectivity of sigma38-containing RNA polymerase is observed only under specific reaction conditions. The examination of 6S RNA, which is encoded by the ssr1 gene in vivo, showed that it reduced Esigma70 activity during the stationary phase, but this reduction of activity did not result in the elevation of Esigma38 activity. Thus, the preferential expression of stationary-phase genes by Esigma38 is unlikely the consequence of selective inhibition of Esigma70 by 6S RNA. PMID- 15357304 TI - Growth inhibition of Escherichia coli during heterologous expression of Bacillus subtilis glutamyl-tRNA synthetase that catalyzes the formation of mischarged glutamyl-tRNA1 Gln. AB - It is known that Bacillus subtilis glutamyl-tRNA synthetase (GluRS) mischarges E. coli tRNA1 Gln with glutamate in vitro. It has also been established that the expression of B. subtilis GluRS in Escherichia coli results in the death of the host cell. To ascertain whether E. coli growth inhibition caused by B. subtilis GluRS synthesis is a consequence of Glu-tRNA1 Ghn formation, we constructed an in vivo test system, in which B. subtilis GluRS gene expression is controlled by IPTG. Such a system permits the investigation of factors affecting E. coli growth. Expression of E. coli glutaminyl-tRNA synthetase (GlnRS) also ameliorated growth inhibition, presumably by competitively preventing tRNA1 Gln misacylation. However, when amounts of up to 10 mM L-glutamine, the cognate amino acid for acylation of tRNA1 Gln, were added to the growth medium, cell growth was unaffected. Overexpression of the B. subtilis gatCAB gene encoding Glu-tRNAGln amidotransferase (Glu-AdT) rescued cells from toxic effects caused by the formation of the mischarging GluRS. This result indicates that B. subtilis Glu AdT recognizes the mischarged E. coli GlutRNA1 Gln, and converts it to the cognate Gln-tRNA1 Gln species. B. subtilis GluRS-dependent Glu-tRNA1 Gln formation may cause growth inhibition in the transformed E. coli strain, possibly due to abnormal protein synthesis. PMID- 15357305 TI - Antibody response to crude cell lysate of propionibacterium acnes and induction of pro-inflammatory cytokines in patients with acne and normal healthy subjects. AB - Propionibacterium acnes (P. acnes) plays an important role in the disease pathogenesis of acne vulgaris, a disorder of pilosebaceous follicles, seen primarily in the adolescent age group. In the present study, the presence of antibodies against P. acnes (MTCC1951) were detected in acne patient (n=50) and disease free controls (n=25) using dot-ELISA and Western blot assay. The ability of P. acnes to induce pro-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs), obtained from acne patients and healthy subjects, were also analysed. The patients (n=26) who were culture positive for skin swab culture, were found to have a more advanced disease and higher antibody titres (1:4000 to > 1:16000) compared to the P. acnes negative patients (n=24) and normal controls (n=25). An analysis of patients' sera by western blot assay recognized a number of antigenic components of P. acnes, ranging from 29 to 205 kDa. The major reactive component was an approximately 96 kDa polypeptide, which was recognised in 92% (24 of 26) of the patients sera. Further, the P. acnes culture supernatant, crude cell lysate and heat killed P. acnes whole cells, obtained from 72-h incubation culture, were observed to be able to induce significant amounts of IL-8 and tumor necrosis factor alpha (TNF-alpha) by the PBMCs in both the healthy subjects and patients, as analysed by cytokine-ELISA. The levels of cytokines were significantly higher in the patients than the healthy subjects. A major 96 kDa polypeptide reactant was eluted from the gel and was found to cause dose dependent stimulation of the productions of IL-8 and TNF alpha. Thus, the above results suggest that both humoral and pro-inflammatory responses play major roles in the pathogenesis of acne. PMID- 15357306 TI - Expression and purification of a recombinant scFv towards the exotoxin of the pathogen, Burkholderia pseudomallei. AB - A single chain variable fragment (scFv) specific towards B. pseudomallei exotoxin had previously been generated from an existing hybridoma cell line (6E6AF83B) and cloned into the phage display vector pComb3H. In this study, the scFv was subcloned into the pComb3X vector to facilitate the detection and purification of expressed antibodies. Detection was facilitated by the presence of a hemagglutinin (HA) tag, and purification was facilitated by the presence of a histidine tag. The culture was grown at 30 degrees C until log phase was achieved and then induced with 1 mM IPTG in the absence of any additional carbon source. Induction was continued at 30 degrees C for five h. The scFv was discerned by dual processes-direct enzyme-linked immunosorbent assays (ELISA), and Western blotting. When compared to E. coli strains ER2537 and HB2151, scFv expression was observed to be highest in the E. coli strain Top10F'. The expressed scFv protein was purified via nickel-mediated affinity chromatography and results indicated that two proteins a 52 kDa protein, and a 30 kDa protein were co-purified. These antibodies, when blotted against immobilized exotoxin, exhibited significant specificity towards the exotoxin, compared to other B. pseudomallei antigens. Thus, these antibodies should serve as suitable reagents for future affinity purification of the exotoxin. PMID- 15357307 TI - Free fatty acid accumulation by mesophilic lactic acid bacteria in cold-stored milk. AB - This study was aimed to determine the accumulation of free fatty acid by mesophilic lactic acid bacteria (Lactococcus lactis subsp. lactis 1471, Lactococcus lactis subsp. cremoris 1000 and Lactobacillus casei 111) in cold stored milk. According to the results, all cold-stored milks had higher acid degree values than those of fresh milk. This phenomenon showed that a slight increase occurred in the accumulation of free fatty acids as a result of spontaneous lipolysis during cold storage. All lactic acid bacteria showed good performance in production of titratable acidity, which increased during fermentation of the milk (fresh and stored milks). Moreover, as the storage time was prolonged, more free fatty acid accumulation was obtained from the fermentation of the cold-stored milk by the investigated lactic acid bacteria. The control milk, which was without lactic acid bacteria, showed no change in the accumulation of free fatty acid during fermentation. From this result, it can be suggested that longer cold-storage time can induce higher free fatty acid accumulation in milk by lactic acid bacteria. PMID- 15357308 TI - Isolation of Citrobacter sp. mutants defective in decolorization of brilliant green by transposon mutagenesis. AB - To identify genes involved in the decolorization of brilliant green, we isolated random mutants generated by transposon insertion in brilliant green-decolorizing bacterium, Citrobacter sp. The resulting mutant bank yielded 19 mutants with a complete defect in terms of the brilliant green color removing ability. Southern hybridization with a Tn5 fragment as a probe showed a single hybridized band in 7 mutants and these mutants appeared to have insertions at different sites of the chromosome. Tn5-inserted genes were isolated and the DNA sequence flanking Tn5 was determined. By comparing these with a sequence database, putative protein products encoded by bg genes were identified as follows: bg 3 as a LysR-type regulatory protein; bg 11 as a MalG protein in the maltose transport system; bg 14 as an oxidoreductase; and bg 17 as an ABC transporter. The sequences deduced from the three bg genes, bg 2, bg 7 and bg 16, showed no significant similarity to any protein with a known function, suggesting that these three bg genes may encode unidentified proteins responsible for the decolorization of brilliant green. PMID- 15357310 TI - Identification of sigmaB-dependent promoters using consensus-directed search of Streptomyces coelicolor genome. AB - SigmaB plays an important role in both osmoprotection and proper differentiation in Streptomyces coelicolor A3(2). We searched for candidate members of the sigmaB regulon from the genome database, using the consensus promoter sequence (GNNTN14 16GGGTAC/T). The list consists of 115 genes, and includes all the known sigmaB target genes and many other genes whose functions are related to stress protection and differentiation. PMID- 15357309 TI - DD1.5k, the gene preferentially expressed in bloodstream isolates of vancomycin resistant Enterococcus faecium. AB - Vancomycin-resistant Enterococcus faecium (VREFM) is becoming a threatening pathogen. We identified a gene called DD1.5K by differential display-PCR, which was preferentially expressed in the bloodstream isolates of VREFM. Due to its amino acid similarity to transfer complex protein, trsE, and tissue-specific expression, this gene may be involved in virulence of VREFM. PMID- 15357311 TI - Characterization of protocatechuate 4,5-dioxygenase induced from p hydroxybenzoate-cultured Pseudomonas sp. K82. AB - Pseudomonas sp. K82 has been reported to be an aniline-assimilating soil bacterium. However, this strain can use not only aniline as a sole carbon and energy source, but can also utilize benzoate, p-hydroxybenzoate, and aniline analogues. The strain accomplishes this metabolic diversity by using different aerobic pathways. Pseudomonas sp. K82, when cultured in p-hydroxybenzoate, showed extradiol cleavage activity of protocatechuate. In accordance with those findings, our study attempted the purification of protocatechuate 4,5-dioxygenase (PCD 4,5). However the purified PCD 4,5 was found to be very unstable during purification. After Q-sepharose chromatography was performed, the crude enzyme activity was augmented by a factor of approximately 4.7. From the Q-sepharose fraction which exhibited PCD 4,5 activity, two subunits of PCD4,5 (alpha subunit and beta subunit) were identified using the N-terminal amino acid sequences of 15 amino acid residues. These subunits were found to have more than 90% sequence homology with PmdA and PmdB of Comamonas testosteroni. The molecular weight of the native enzyme was estimated to be approximately 54 kDa, suggesting that PCD4,5 exists as a heterodimer (alpha1beta1). PCD 4,5 exhibits stringent substrate specificity for protocatechuate and its optimal activity occurs at pH 9 and 15 degrees C. PCR amplification of these two subunits of PCD4,5 revealed that the alpha subunit and beta subunit occurred in tandem. Our results suggest that Pseudomonas sp. K82 induced PCD 4,5 for the purpose of p-hydroxybenzoate degradation. PMID- 15357312 TI - The influence of NaCl and carbonylcyanide-m-chlorophenylhydrazone on the production of extracellular proteases in a marine Vibrio strain. AB - In general, the salinity of the ocean is close to 3.5% and marine vibrios possess the respiratory chain-linked Na+ pump. The influence of sodium chloride and the proton conductor carbonylcyanide m-chlorophenylhydrazone (CCCP) on the production of extracellular proteases in a marine Vibrio strain was examined. At the concentration of 0.5 M, sodium chloride minimally inhibited the activity of extracellular proteases by approximately 16%, whereas at the same concentration, the producton of extracellular proteases was severely inhibited. On the other hand, the production of extracellular proteases was completely inhibited by the addition of 2 microM CCCP at pH 8.5, where the respiratory chain-linked Na+ pump functions. PMID- 15357313 TI - Molecular cloning and identification of a novel oxygenase gene specifically induced during the growth of Rhodococcus sp. strain T104 on limonene. AB - Rhodococcus sp. strain T104 is able to utilize both limonene and biphenyl as growth substrates. Furthermore, T104 possesses separate pathways for the degradation of limonene and biphenyl. Previously, we found that a gene(s) involved in limonene degradation was also related to indigo-producing ability. To further corroborate this observation, we have cloned and sequenced a 8,842-bp genomic DNA region with four open reading frames, including one for indole oxygenase, which converts indole to indigo (a blue pigment), The reverse transcription PCR data demonstrated that the identified indole oxygenase gene is specifically induced by limonene, thereby implicating this gene in the degradation of limonene by T104. PMID- 15357314 TI - Distribution of lichen flora on South Korea. AB - After an overview on the temporary situation of the lichenology in South Korea, localities of 95 macrolichen taxa are reported for South Korea. In this revised lichen flora of South Korea, 16 species are apparently new to the territory. Voucher specimens have been deposited in the Korean Lichen Research Institute (KoLRI) at Sunchon National University in Korea, and duplicates have also been donated to the National History Museum and Institute, in Chiba, (CBM) Japan. PMID- 15357315 TI - The genetic immunodeficiency disease, leukocyte adhesion deficiency, in humans, dogs, cattle, and mice. AB - This review highlights the genotype-phenotype relationship of the genetic immunodeficiency disease leukocyte adhesion deficiency (LAD) in humans, dogs, cattle, and mice, and provides assessment of the opportunities that each animal species provides in the understanding of leukocyte biology and in developing new therapeutic approaches to LAD in humans. This comparison is important since animal models of genetic diseases in humans provide the opportunity to test new therapeutic approaches in an appropriate, disease-specific model. The success of this approach is dependent on the relationship of the phenotype in the animal to the phenotype of the disease in humans. PMID- 15357316 TI - Genetic resistance to chemical hepatocarcinogenesis in the DRH rat strain. AB - The carcinogen-resistant inbred rat strain DRH established from closed-colony Donryu rats by use of selective brother-sister mating over 20 generations under continuous feeding of 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) maintains a highly resistant phenotype without carcinogen exposure for many years. We reported that the clonal expansion of preneoplastic glutathione S-transferase P(GST-P)-positive foci induced by 3'-Me-DAB was less extensive in the liver of DRH rats than in the liver of susceptible strains, such as Donryu and F344, although levels of DNA adducts were comparable among these rats. Comparative studies of the events after initiation indicate that DRH rats are constitutionally less prone to cellular damage caused by continuous administration of 3'-Me-DAB than are parental Donryu rats. Consequently, the reduced growth response of the liver during the promotion stage may contribute to the low susceptibility to development of liver tumors. Genetic analysis of (F344 x DRH)F2 rats identified two quantitative trait loci, Drh1 on chromosome 1 and Drh2 on chromosome 4, which provide resistance to the development of GST-P positive preneoplastic foci induced by 3'-Me-DAB during the early stage of its administration. The resistance to progression to hepatocellular carcinoma is affected solely by Drh2. These observations indicate that at least two genetic loci are critically involved in the steps leading to chemical hepatocarcinogenesis. The DRH rat is a useful experimental model with which to study genetic susceptibility and resistance to chemically induced liver cancers. PMID- 15357317 TI - Comparison of the effects of perfusion in determining brain penetration (brain-to plasma ratios) of small molecules in rats. AB - In the process of drug discovery, brain and plasma measurements of new chemical entities in rodents are of interest, particularly when the target receptors are in the brain. Brain-to-plasma ratios (B/P) obtained from a rodent pharmacokinetic assay are useful in helping determine which compounds are brain penetrant. The study reported here was performed to determine whether whole-body saline perfusion for complete blood removal was required to accurately measure brain tissue compound concentrations. Diazepam was used as a positive control since it is highly brain penetrant. Compound A was used as a negative control since it had known poor brain penetration. After intravenous dosing with either diazepam or compound A, rats were anesthetized and blood was collected, then the brain was removed following no perfusion or whole-body perfusion with saline. The analytes described (compound A, diazepam, and the internal standard) were recovered from plasma or brain homogenate by use of protein precipitation, and were subsequently analyzed by use of liquid chromatography/tandem mass spectrometry (LC/MS/MS). The B/P values determined by use of LC-MS were not significantly different in perfused vs. non-perfused rats (P > or = 0.05). This approach (whole brain collected from non-perfused male rats) is an attractive alternative over brain penetration studies of perfused rats, since it has markedly reduced the technical time and potential for pain and distress required for generating B/P data due to elimination of the requirement for anesthesia and surgical preparation of animals. PMID- 15357318 TI - Efficacy of three microbiological monitoring methods in a ventilated cage rack. AB - The use of individually ventilated caging (IVC) to house mice presents new challenges for effective microbiological monitoring. Methods that exploit the characteristics of IVC have been developed, but to the authors' knowledge, their efficacy has not been systematically investigated. Air exhausted from the IVC rack can be monitored, using sentinels housed in cages that receive rack exhaust air as their supply air, or using filters placed on the exhaust air port. To aid laboratory animal personnel in making informed decisions about effective methods for microbiological monitoring of mice in IVC, the efficacy of air monitoring methods was compared with that of contact and soiled bedding sentinel monitoring. Mice were infected with mouse hepatitis virus (MHV), mouse parvovirus (MPV), murine rotavirus (agent of epizootic diarrhea of mice [EDIM]), Sendai virus (SV), or Helicobacter spp. All agents were detected using contact sentinels. Mouse hepatitis virus was effectively detected in air and soiled bedding sentinels, and SV was detected in air sentinels only. Mouse parvovirus and Helicobacter spp. were transmitted in soiled bedding, but the efficacy of transfer was dependent on the frequency and dilution of soiled bedding transferred. Results were similar when the IVC rack was operated under positive or negative air pressure. Filters were more effective at detecting MHV and SV than they were at detecting MPV. Exposure of sentinels or filters to exhaust air was effective at detecting several infectious agents, and use of these methods could increase the efficacy of microbiological monitoring programs, especially if used with soiled bedding sentinels. In contemporary mouse colonies, a multi-faceted approach to microbiological monitoring is recommended. PMID- 15357319 TI - Comparative study of lung cytologic features in normal rhesus (Macaca mulatta), cynomolgus (Macaca fasicularis), and African green (Chlorocebus aethiops) nonhuman primates by use of bronchoscopy. AB - Invasive bronchoscopy and bronchoaveolar lavage (BAL) fluid collection represents an important tool in studies of the respiratory system of nonhuman primates. Bronchoscopy and BAL fluid collection was performed on groups of rhesus (Macaca mulatta) and cynomolgus (Macaca fasicularis) macaques and African green monkeys (Chlorocebus aethiops), and the resulting comparative lavage cytologic features are described. Analysis of the BAL fluid did not reveal significant differences among species with respect to total cells recovered or differential cellular composition. This description of the method used to lavage the nonhuman primates and the resulting lung cytologic findings provide important comparative data for three species commonly used in biomedical research. PMID- 15357320 TI - Effect of isoflurane, atracurium, fentanyl, and noxious stimulation on bispectral index in pigs. AB - The study reported here was done to determine the relationship between anesthesia depth and bispectral index (BIS) in stimulated pigs. Isoflurane minimal alveolar concentration (MAC) was determined using the tail-clamp method in 16 Yorkshire/Landrace-cross pigs with mean+/-SEM weight of 27.7+/-1.76 kg. One week later, BIS, ECG, heart rate, arterial blood pressure, esophageal temperature, end tidal CO2 tension and isoflurane concentration, arterial pH, PaO2, PaCO2, plasma bicarbonate concentration, and base excess were determined at each of five isoflurane MAC-multiples: 0.8, 1.0, 1.3, 1.6, and 2.0. Six treatments were studied: isoflurane; isoflurane and atracurium; isoflurane, atracurium, and fentanyl; isoflurane with noxious stimulation; isoflurane and atracurium with noxious stimulation; and isoflurane, atracurium, and fentanyl with noxious stimulation. The noxious stimulus during BIS measurement was the same as that for MAC determination. Each pig was studied three times (n = 8), and order of MAC multiples and treatments was randomized. Data were evaluated by use of general linear model analysis of variance and linear regression analysis, with statistical significance set at P < 0.05. Significant differences in BIS values were identified between MAC-multiples within each treatment and between treatment 3 compared with treatments 2 and 4. Significant differences also were observed within and between treatments for heart rate, arterial blood pressure, and PaO2. Use of BIS appears reliable for identification of light versus deep anesthesia, but is of limited use for discrimination between isoflurane MAC-multiples of 1 and 1.6. We conclude that, compared with other treatments, atracurium and noxious stimulation had no significant effect on BIS. PMID- 15357321 TI - Mycoplasma haemocanis infection--a kennel disease? AB - Mycoplasma haemocanis (formerly Haemobartonella canis) is a red blood cell parasite that causes disease mainly in immunosuppressed and splenectomized dogs. Clinical outbreak of the disease resulted in failure of a large experimental project. We aimed to identify whether M. haemocanis has increased prevalence in kennel-raised dogs. In a prospective study, we compared the prevalence of M. haemocanis in whole blood (anti-coagulated by use of EDTA) collected from pet dogs (University of Illinois, Urbana Champaign, Ill.; n = 60) with that in blood from dogs raised in three distinct kennels in western Europe (WE; n = 23), eastern Europe (EE; n = 20), and North America (NA; n = 20). Screening included antibody testing and microscopy of blood smears. The presence of M. haemocanis was identified using a polymerase chain reaction (PCR) assay for specific DNA of the organism. None of the pet dogs (0%) was test positive for M. haemocanis DNA. Mycoplasma haemocanis was found in dogs tested at all of the kennels. Infection rate in the three kennels was 30, 35, and 87%, respectively (all P < 0.001 versus control, chi2-test). Latent infection with M. haemocanis was not a single observation in kennel-raised dogs. Prevalence may be higher than that in a pet dog population. The potential exists for these latent infections to adversely affect or confound research results. PMID- 15357322 TI - Reverse transcription-polymerase chain reaction detection and nucleic acid sequence confirmation of reovirus infection in laboratory mice with discordant serologic indirect immunofluorescence assay and enzyme-linked immunosorbent assay results. AB - Reovirus infections are typically subclinical in weaned mice, and are best detected using serologic tests. After exposure to the soiled bedding of some mice obtained from various sources, numerous sentinel mice tested reovirus seropositive by use of indirect immunofluorescence assays (IIFA) in our institution. A major commercial rodent pathogen testing laboratory verified our IIFA results, but since the same samples were reovirus seronegative using their "more specific" enzyme-linked immunosorbent assay (ELISA), the IIFA results were reported as "false positives." As past in-house observations suggested transmission of the virus to sentinel and other animals, we sought to determine whether the IIFA results were always "false positives." An opportunity to test this notion arose after receipt of reovirus IIFA-positive transgenic mice from an academic source. Using reverse transcriptase-polymerase chain reaction (RT-PCR) assays, the presence of reovirus RNA was detected in fecal specimens taken from some sentinel animals that subsequently seroconverted from IIFA-negative to IIFA positive for reovirus. The virus could not be isolated by use of tissue culturing methods. Nucleotide sequence analysis established the presence of unique reovirus sequences. These results indicate that contemporary reovirus infections may not be detected by use of some serologic tests, and that RT-PCR analysis may be useful for confirmation of active reovirus infection in certain situations. PMID- 15357323 TI - Increased incidence of vaginal septum in C57BL/6J mice since 1976. AB - Decreased fertility was observed in a breeding colony of C57BL/6J mice. On examination, a dorsoventral vaginal septum was detected in many females. This defect was identified in 1976, with incidence of 4.0% in this strain. Our objective was to determine whether incidence of this condition has increased and whether this defect was associated with the observed infertility. We report incidence of 11.3%, nearly triple the original reported incidence. For comparison, incidence of vaginal septum in C57BL/6N females was determined and was found to be 1%. We performed a breeding study using normal and affected C57BL/6J females to evaluate fertility in affected females. Our data were consistent with those of the 1976 report; fertility was decreased in females with an intact vaginal septum. In 50% of affected females, the septum remained intact after breeding. The fertility for this subgroup of vaginal septum-retained females was 14.3%, compared with 85.7% in females whose septum ruptured and 75.0% in normal females (statistically significant, P = 0.02). On the basis of our results, we provide animal and financial loss data due to the defect. Lastly, we provide suggestions on how to minimize animal losses and be in accordance with the principles of the 3Rs (replacement, refinement, reduction). PMID- 15357324 TI - Subacute necrotizing encephalopathy in a pig-tailed macaque (Macaca nemestrina) that resembles mitochondrial encephalopathy in humans. AB - A male pig-tailed macaque (Macaca nemestrina), approximately 5 years old, was found to be vision-impaired and to have profound behavioral abnormalities, including hyperactivity and self-injurious behavior that was not amenable to amelioration by environmental enrichment. Facial and skeletal dysmorphisms also were noted. Magnetic resonance imaging (MRI) and positron emission tomography (PET) scanning revealed areas of possible infarction in the occipital lobes and megaventriculosis. At necropsy, following euthanasia for humane reasons, severe polio- and leukoencephalomalacia accompanied by megaventriculosis were seen in both occipital lobes and in several sulci of the parietal and frontal lobes. Light microscopic findings included loss of neocortical structure, with necrosis, neuronal loss, astrogliosis, vascular proliferation, mild spongiosis, and demyelination. The extent and severity of lesions were most pronounced in the occipital lobes and were greater in the left than in the right hemisphere. Other lesions included mild bilateral atrophy of the optic nerves, thymic involution, necrotizing dermatitis due to trauma, and a spectrum of spermatozoal abnormalities. The imaging and gross and light microscopic changes found in this animal resemble the mitochondrial encephalopathies of humans; this was corroborated by results of immunohistochemical analysis demonstrating decreased expression of enzymes of the mitochondrial oxidative complex ([OC]-I, -III, and IV) in brain and muscle, and detection of fibrinogen immunoreactivity in neurons and glial cells. The spermatozoal defects may represent yet another aspect of a mitochondrial defect. PMID- 15357325 TI - Outbreak of sarcoptic mange and malasseziasis in rabbits (Oryctolagus cuniculus). AB - An outbreak of combined Sarcoptes and Malassezia spp. infection was diagnosed in a rabbitry. About 20 (4%) of 500 rabbits were affected. Two 6- to 8-month-old female Holland Lops rabbits were submitted to the Tifton Diagnostic & Investigational Laboratory at The University of Georgia for complete necropsy. Gross lesions consisted of marked multifocal areas of alopecia, crusting, and dermatitis around the eye and on ears, nose, lips, neck, abdomen, feet, and external genitalia. Histologic examination of the skin revealed epidermal acanthosis with marked parakeratotic hyperkeratosis and cross sections of embedded mites consistent with Sarcoptes sp. and budding yeasts consistent with Malassezia sp. To the best of the author's knowledge, this is the first case report of combined Sarcoptes and Malassezia spp. infection in rabbits. PMID- 15357326 TI - Ampullary carcinoma in a group of aged rhesus macaques (Macaca mulatta). AB - Ampullary carcinoma was diagnosed in 6 rhesus macaques that ranged in age from 20 to 35 years. Signalment, premonitory signs of disease, and results of clinical biochemical and hematologic analyses varied among animals. Histologically, the neoplastic cells obliterated the ampulla, with regional spread to the duodenum in all 6 animals and to the pancreas in one animal. Two animals had metastases to the lung, and another two had metastases to the pancreoduodenal lymph nodes and liver. One animal had mesocolonic metastasis. Malignant tumors of the ampullary region are rare in domestic animals, and account for less than 5% of all cancers of the digestive tract in humans. PMID- 15357327 TI - Lacrimal gland myoepithelioma in a rhesus macaque (Macaca mulatta). AB - An 18-year-old rhesus macaque (Macaca mulatta) developed ptosis of the left upper eyelid due to a mass that had first been observed 10 years previously. The 11 x 7 x 7-mm mass was surgically excised, and the ptosis resolved after 5 days. Histologic examination of the mass revealed two confluent cell populations. Most cells were spindle-shaped and were arranged in loose fascicles. Smaller numbers of cells had squamous differentiation. The spindle-shaped cells expressed smooth muscle actin. Cells with squamous differentiation did not express smooth muscle actin, but did, along with around half of the spindle-shaped cells, express pan cytokeratin. On the basis of histologic and immunohistochemical findings, the mass was diagnosed as myoepithelioma. The neoplasm most likely originated from the palpebral lobe of the lacrimal gland, although accessory lacrimal gland origin could not be excluded. Recurrence of the neoplasm has not been observed 6 months after surgery. PMID- 15357328 TI - Life course transitions and depressive symptoms among women in midlife. AB - This study examined the relationship between three midlife transitions and depressive symptoms among 952 women 50 to 59 years of age. Using longitudinal data from women interviewed for the 1992 and 2000 Health and Retirement Study, the study described changes in marital status, change to a parental caregiving role, and changes in perceived health across the eight years. Further, it examined the impact of these changes on mental health. The findings indicate that becoming widowed, becoming a caregiver, and perceiving health declines significantly increased depressive symptoms in the year 2000, even when controlling for pre-transition levels of depressive symptoms. The findings are consistent with the lifecourse perspective that individual development occurs in context and across the lifespan. The findings confirm and add to current midlife research literature. PMID- 15357329 TI - Expressions of generativity and posttraumatic growth in adult cancer survivors. AB - Much of the psycho-oncology research that has been conducted to date has focused on understanding the negative psychological and psychosocial sequelae of cancer. However, a growing body of evidence suggests that many cancer survivors report psychological growth following a diagnosis of cancer. Further, there are few studies that examine the relationship among age, cancer, and posttraumatic growth. This study examines relationships among posttraumatic growth, generativity, age, and gender. Questionnaires were mailed to 400 randomly selected cancer survivors from a metropolitan hospital tumor registry, with a yield of 21%. Three age cohorts were examined (26-41 years, 42-54 years, and 55 plus years). As anticipated, MANOVA showed no significant age differences in expression of generative concern among the three cohorts, but there was evidence that younger adults engage in a greater number of generative behaviors than older adults. Further, females reported more positive growth on four out of the five domains of the measure of posttraumatic growth compared to males, and the oldest cohort experienced lower levels of positive growth compared with the other two groups. Generative concern was positively correlated with the total posttraumatic growth score. Implications for theoretical and applied research on generativity and posttraumatic growth in adult cancer survivors are discussed. PMID- 15357330 TI - Adulthood predictors of health promoting behavior in later aging. AB - This study investigated adulthood predictors of health-promoting behavior in later aging. The participants were 162 members of the Terman Study of the Gifted (Terman et al., 1925), who responded in 1999 at an average age of 86 to a mailout questionnaire which included questions concerning their positive health behavior. Adulthood variables were obtained for analysis from the study archives. Cross sectional analyses showed that health-promoting behavior was related to perceived health and well-being in later aging. In longitudinal analyses, "easygoingness" in adulthood was positively related to positive psychosocial behavior and positive health practices, and neuroticism was negatively related to positive psychosocial behavior in later aging. Perceived health in adulthood was related to positive psychosocial behavior, exercise, physical recreation, and other positive health practices. Education, perceived health in adulthood, recalled importance in adulthood of success in sports before the age of 12, and interest in outdoor sports in early adulthood, as well as current health problems, were related to exercise and physical recreation in later aging. Gender played a moderating role in the prediction of positive psychosocial behavior, with the relationship of self-rated health in adulthood and recalled importance of success in sports before the age of 12 with positive psychosocial behavior being stronger for men than for women. PMID- 15357331 TI - Stereotypes of the elderly in U.S. television commercials from the 1950s to the 1990s. AB - Critics have charged that American advertisers have often portrayed the elderly with negative stereotypes. These negative portrayals, they suggest, not only offend elderly consumers but also contribute to ageism. This study examined whether American advertisers have indeed used a great deal of negative stereotyping of the elderly, as the critics have suggested. Employing a concept of stereotypes found in cognitive psychology, the authors examined trends in the portrayal of the elderly in television commercials produced in the United States from the 1950s through the 1990s. Results of the study do not support the contentions of the critics. Very little negative stereotyping was discovered. Analysis indicated trends in the appearance of several positive stereotypes, and differences in the stereotyping of gender groups and age segments within the elderly group. Results are interpreted from both marketing and social science perspectives. PMID- 15357332 TI - Application of a widely electrically tunable diode laser to chemical gas sensing with quartz-enhanced photoacoustic spectroscopy. AB - A near-infrared diode laser with sample-grating distributed Bragg reflectors was used as a widely tunable spectroscopic source for multispecies chemical sensing. Quartz-enhanced photoacoustic spectroscopy was utilized to obtain high absorption sensitivity in a compact gas cell. CO2, H2O C2H2, and NH3 were monitored. A noise equivalent sensitivity of 8 x 10(-9) cm(-1) W(-1) Hz(-1/2)for NH3 detection was achieved, which corresponds to a NH3 mixing ratio of 4.4 parts in 10(6) by volume (ppmv) with a 1-s time constant and available 5.2-mW optical power in the gas cell. PMID- 15357333 TI - Femtosecond laser fabrication of tubular waveguides in poly(methyl methacrylate). AB - Femtosecond laser direct writing is employed for the fabrication of buried tubular waveguides in bulk poly(methyl methacrylate). A novel technique using selective chemical etching is presented to resolve the two-dimensional refractive index profile of the fabrication structures. End-to-end coupling in the waveguides reveals a near-field intensity distribution that results from the superimposition of several propagating modes with different azimuthal symmetries. Mode analysis of the tubular waveguides is performed using the finite-difference method, and the possible propagating mode profiles are compared with the experimental data. PMID- 15357334 TI - Dual-pump dual-broadband coherent anti-Stokes Raman scattering in reacting flows. AB - A dual-pump, dual-broadband coherent anti-Stokes Raman scattering system for simultaneous measurements of temperature and concentrations of N2, O2, and CO2 in reacting flows is demonstrated. In this system pure rotational transitions of N2 O2 and rovibrational transitions of N2-CO2 are probed simultaneously with two narrowband pump beams, a broadband pump beam, and a broadband Stokes beam. The main advantage of this technique is that it permits accurate temperature measurements at both low and high temperatures as well as concentration measurements of three molecules. PMID- 15357335 TI - Multilevel phase-type diffractive lenses in silica glass induced by filamentation of femtosecond laser pulses. AB - Multilevel phase-type diffractive lenses were fabricated by translating a filament of a femtosecond laser pulse into three-dimensional space inside synthesized silica. The profile of the lenses was designed by use of a multi level approximation to a kinoform lens. Two-level diffractive lenses with multiple layers along the optical axis provided a maximum efficiency of 37.6% at a wavelength of 632.8 nm. A four-level diffractive lens provided a maximum efficiency of 56.9%. The lenses fabricated with filamentation were free from birefringence. PMID- 15357336 TI - Simple analysis of the energy density distribution of the diffracted ultraviolet beam from a fiber Bragg grating phase mask. AB - The energy density distribution of the diffracted ultraviolet field from a phase mask used for fiber Bragg grating fabrication is efficiently analyzed at short-to intermediate distances by application of a thin phase screen model and the angular spectrum decomposition method. A direct comparison with the results obtained from a rigorous coupled-wave model shows good agreement. PMID- 15357337 TI - Destruction of silica fiber cladding by the fuse effect. AB - The phenomenon of destruction of silica fiber cladding by the fiber fuse effect has been observed for the first time to the authors' knowledge. Experiments on the optical discharge propagation along a fiber were conducted with fibers of decreased cladding thickness. The destruction of fiber cladding led to expansion of the optical discharge plasma and to a decrease of its density. This resulted in the termination of optical discharge propagation. The section of a fiber with decreased cladding thickness can act as a safety device to halt damage propagation. PMID- 15357338 TI - Single-polarization fiber with a high extinction ratio. AB - An elliptical-core hole assisted single-polarization fiber was designed, fabricated, and characterized. Numerical modeling based on the vectorial Maxwell equation reveals the dependence of the single-polarization bandwidth on core delta and air-hole size. Several single-polarization fibers based on this design with their single-polarization operating windows centered between 0.9 and 1.5 microm were successfully demonstrated. A correlation between fiber birefringence and single-polarization operating bandwidth is qualitatively confirmed. A single polarization bandwidth as high as 55 nm was observed. These fibers also show very high extinction ratios of 60 dB or higher at lengths much shorter than 1 m. Other properties such as the dependence on length of the single-polarization operating window were also measured. PMID- 15357339 TI - Fused preforms for the fabrication of photonic crystal fibers. AB - A novel approach for the fabrication of high-quality preforms for use in the fabrication of photonic crystal fibers is described. The preforms are fabricated in a multistep process that involves stacking a bundle with rods and (or) tubes of two dissimilar glasses, fusing the bundle, and then etching the fused bundle in acid to remove one of the two glasses. The procedure for fabrication of the fused preforms is similar to that used in the fabrication of microchannel plate glass and yields periodically spaced, uniform, round channels that extend through the length of the preform. PMID- 15357340 TI - Optical packet labeling based on simultaneous polarization shift keying and amplitude shift keying. AB - We propose and demonstrate label encoding-swapping and transmission for an orthogonally labeled packet by use of a 10-Gbit/s amplitude shift keying payload and a 2.5-Gbit/s polarization shift keying (PolSK) label. A simple scheme for demodulating and demultiplexing the PolSK label based on an injection-locked Fabry-Perot laser diode is described. The extinction ratio of the newly added PolSK label can be precisely controlled and maintained after the intermediate node. PMID- 15357341 TI - Pulse shaper in a loop: demonstration of cascadable ultrafast all-optical code translation. AB - We experimentally demonstrate repetitive M-ary spectral phase pulse shaping by placing a programmable pulse shaper driven by a 10-GHz source in a closed loop. This permits generation of encoded and decoded signals in the same apparatus by forming a closed loop to circulate a part of the output back into the pulse shaper. As a result, a series of M - 1 distinct encoded waveforms is sequentially generated, followed by generation of a properly decoded pulse. PMID- 15357342 TI - Tunable diffraction of magnetic fluid films and its potential application in coarse wavelength-division multiplexing. AB - When an external magnetic field is applied parallel to the film surface of a magnetic fluid film, a high-quality one-dimensional periodic chain structure is formed when the field strength reaches a certain level. With a periodic chain structure in the magnetic fluid film, an incident light is diffracted onto the magnetic thin film. The results show that the one-dimensional periodic chain structure in the magnetic fluid film can serve as an optical grating. Further investigations reveal the feasibility of developing tunable coarse wavelength division multiplexing by utilizing a periodic chain structure. PMID- 15357343 TI - Observation of Ince-Gaussian modes in stable resonators. AB - We report what is to our knowledge the first observation of Ince-Gaussian modes directly generated in a stable resonator. By slightly breaking the symmetry of the cavity of a diode-pumped Nd:YVO4 laser and its pump beam configuration we were able to generate single high-order Ince-Gaussian modes of high quality. The observed transverse modes have an inherent elliptic structure and exhibit remarkable agreement with theoretical predictions. PMID- 15357344 TI - Flow imaging by use of femtosecond-laser-induced two-photon fluorescence. AB - A novel technique is demonstrated for the imaging of turbulent flows in which a single window to the flow is the only optical access required. A femtosecond laser is used to excite two-photon fluorescence in a disodium-fluorescein-seeded water jet. The fluorescence signal is generated at only the focal point of the laser because of the highly nonlinear nature of the two-photon absorption, and it is collected in a direction counterpropagating to the excitation beam. Tight focusing of the laser is used to limit the probe volume, and the two-dimensional mean and rms concentration images are collected by raster scanning the laser. PMID- 15357345 TI - Solid-state laser intensity stabilization at the 10(-8) level. AB - A high-power, low-noise photodetector, in conjunction with a current shunt actuator, is used in an ac-coupled servo to stabilize the intensity of a 10-W cw Nd:YAG laser. A relative intensity noise of 1 x 10(-8) Hz(-1/2) at 10 Hz is achieved. PMID- 15357346 TI - High-power tunable diode-pumped Yb3+:CaF2 laser. AB - Results of diode-pumped cw laser operation of an Yb3+:CaF2 single crystal are reported for what is to our knowledge the first time. With a 5-at.% Yb3+ -doped sample we obtained 5.8-W output power at 1053 nm for 15 W of incident power at 980 nm. The laser wavelength could be tuned from 1018 to 1072 nm, and a small signal gain as high as 1.8 was achieved, showing the great potential of Yb3+:CaF2 as an amplifier medium for femtosecond pulses. PMID- 15357347 TI - Microstructured polymer fiber laser. AB - A microstructured polymer optical fiber doped with Rhodamine 6G dye was fabricated and demonstrated as an optical amplifier and a fiber laser. As an amplifier, the fiber achieved a gain in excess of 30 dB. As a pulsed fiber laser, the fiber exhibited a threshold of 20 microJ, a slope efficiency of 18%, and a lifetime as high as 130,000 shots at 10 Hz. The maximum output energy was 16 microJ. The advantages that such fibers offer lie in the simplicity and flexibility of their fabrication and in their potential for use as compact, tunable solid-state sources. PMID- 15357348 TI - Genetic drive of a laser. AB - Direct production of laser output pulses with arbitrary shapes can be extended to the high-frequency domain by use of optimal driving of the pump power. Genetic algorithms allow us to design the optimal time evolutions of the pump power to counteract the detrimental effect of relaxation oscillations. The method is demonstrated on a Nd3+:YVO4 laser and allows us to produce, e.g., triangular pulses at rates 20 times faster than for proportional modulation. PMID- 15357349 TI - Polarization anisotropy in vertical-cavity semiconductor optical amplifiers. AB - The polarization-dependent gain (PDG) characteristics of a vertical-cavity semiconductor optical amplifier (VCSOA) are measured, and the case of the PDG is determined. It is often assumed that the polarization states of a VCSOA are degenerate because of the circular geometry of the device. This assumption is not true in practice, and it is found that VCSOAs possess a dominant linear polarization state and a small difference in frequency between polarization states. The difference in resonant frequencies causes the PDG of the VCSOA. Measurements of the polarization state show that the cause of the splitting is electro-optic birefringence. PMID- 15357350 TI - Wavelength stabilization and spectrum narrowing of high-power multimode laser diodes and arrays by use of volume Bragg gratings. AB - Spectral line narrowing (by a factor of 8) and stabilization of the emission wavelength (by a factor of 30) of multimode high-power laser diodes and arrays is demonstrated by use of volume Bragg gratings fabricated in high-stability inorganic photorefractive glasses. Applications include stabilization of pump laser diodes and arrays for solid-state lasers and metal-vapor lasers, spin hyperpolarization of noble gases used in medical imaging, and others. PMID- 15357351 TI - Microfluidic sensor based on integrated optical hollow waveguides. AB - A simple integrated optical refractometric sensor based on hollow-core antiresonant reflecting optical waveguides is proposed. The sensor uses the antiresonant reflecting guidance mechanism and permits one to measure the refractive index of a liquid filling the core by simply monitoring the transmitted spectrum. The device has been made with standard silicon technology, and the experimental results confirm numerical simulations performed in one- and two-dimensional geometry. The sensor exhibits a linear response over a wide measurement range (1.3330-1.4450) and a resolution of 9 x 10(-4) and requires a small analyte volume. PMID- 15357352 TI - Disorder-induced losses in photonic crystal waveguides with line defects. AB - A numerical analysis of extrinsic diffraction losses in two-dimensional photonic crystal slabs with line defects is reported. To model disorder, a Gaussian distribution of hole radii in the triangular lattice of airholes is assumed. The extrinsic losses below the light line increase quadratically with the disorder parameter, decrease slightly with increasing core thickness, and depend weakly on the hole radius. For typical values of the disorder parameter the calculated loss values of guided modes below the light line compare favorably with available experimental results. PMID- 15357353 TI - Optical waveguide writing with a diode-pumped femtosecond oscillator. AB - Optical waveguide writing is demonstrated by means of a diode-pumped cavity dumped Yb:glass femtosecond laser oscillator with a pulse energy of 270 nJ at a 166-kHz repetition rate. Waveguides realized on an Er:Yb-doped phosphate glass are almost perfectly mode matched to standard single-mode fibers at 1.55 microm and show a 1.2-dB net gain in a standard telecommunications amplifier setup. Waveguide writing with a compact femtosecond laser oscillator is an important step toward introducing this technique into an industrial context. PMID- 15357354 TI - Effects of structural irregularities on modulational instability phase matching in photonic crystal fibers. AB - The effect of structural irregularities in photonic crystal fibers on scalar and vector modulational instability (MI) processes is studied by numerical simulations and experiments. For an anomalous-dispersion regime pump, variations in core ellipticity as small as 0.5% over length scales of the order of several meters are shown to have a negligible effect on scalar MI, yet they completely suppress vector MI. In contrast, for a normal-dispersion regime pump, vector MI is shown to be robust against such fluctuations. PMID- 15357355 TI - Coherent backscattering spectroscopy. AB - Coherent backscattering (CBS) of light in random media has been previously investigated by use of coherent light sources. Here we report a novel method of CBS measurement that combines low spatial coherence, broadband illumination, and spectrally resolved detection. We show that low spatial coherence illumination leads to an anomalously broad CBS peak and a dramatic speckle reduction; the latter is further facilitated by low temporal coherence detection. Thus CBS can be observed in biological tissue and other media that previously were beyond the reach of conventional CBS measurements. We also demonstrate, for the first time to our knowledge, spectroscopic analysis of CBS. CBS spectroscopy may find important applications in probing random media such as biological tissue in which depth-selective measurements are crucial. PMID- 15357356 TI - Discrete cavity solitons. AB - We derive evolution equations describing light propagation in an array of coupled waveguide resonators and predict the existence of discrete cavity solitons. We identify stable, unstable, and oscillating solitons by varying the coupling strength between the anticontinuous and the continuous limit. PMID- 15357357 TI - Optical parametric oscillation in quasi-phase-matched GaAs. AB - We demonstrate an optical parametric oscillator (OPO) based on GaAs. The OPO utilizes an all-epitaxially-grown orientation-patterned GaAs crystal that is 0.5 mm thick, 5 mm wide, and 11 mm long, with a domain reversal period of 61.2 microm. Tuning either the near-IR pump wavelength between 1.8 and 2 microm or the temperature of the GaAs crystal allows the mid-IR output to be tuned between 2.28 and 9.14 microm, which is limited only by the spectral range of the OPO mirrors. The pump threshold of the singly resonant OPO is 16 microJ for the 6-ns pump pulses, and the photon conversion slope efficiency reaches 54%. We also show experimentally the possibility of pump-polarization-independent frequency conversion in GaAs. PMID- 15357358 TI - High-power diode-pumped actively Q-switched Nd:YVO4 self-Raman laser: influence of dopant concentration. AB - Efficient self-Raman frequency conversion from a diode-pumped actively Q-switched Nd:YVO4 laser at 1064 nm to Stokes emission at 1176 nm is achieved for the first time to the author's knowledge. With a 0.2-at.% Nd:YVO4 crystal, greater than 1.5 W of power at a wavelength of 1176 nm at a repetition rate of 20 kHz was generated with a diode pump power of 10.8 W, corresponding to a conversion of 13.9%. Pulse width and peak power were 18 ns and 4.2 kW, respectively. PMID- 15357359 TI - Stable soliton complexes in two-dimensional photonic lattices. AB - We show that two-dimensional photonic Kerr nonlinear lattices can support stable soliton complexes composed of several solitons packed together with appropriately engineered phases. This may open up new prospects for encoding pixellike images made of robust discrete or lattice solitons. PMID- 15357360 TI - Powerful red-green-blue laser source pumped with a mode-locked thin disk laser. AB - We present a red-green-blue laser source with average powers of 8 W in the red, 23 W in the green, and 10.1 W in the blue. The entire pump power for the nonlinear conversion stages is provided by a single laser oscillator without any amplifier stages. Our system does not require any synchronized cavities, and all nonlinear crystals except one are critically phase matched at room temperature. PMID- 15357361 TI - Non-phase-matched enhancement of second-harmonic generation in multilayer nonlinear structures with internal reflections. AB - Traditional notions of second-harmonic generation rely on phase matching or quasi phase matching to achieve good conversion efficiencies. We present an entirely new concept for efficient second-harmonic generation that is based on the interference of counterpropagating waves in multilayer structures. Conversion efficiencies are an order of magnitude larger than with phase-matched second harmonic generation in similar multilayer structures. PMID- 15357362 TI - Focusing coherent soft-x-ray radiation to a micrometer spot size with an intensity of 10(14) W/cm2. AB - We investigate the focusability of intense coherent soft-x-ray radiation generated by high-order harmonic conversion. The 27th-harmonic wave at 29.6 nm is focused by an off-axis parabolic mirror with a SiC/Mg multilayer coating. Focal spot images are observed from the visible fluorescence induced by the soft-x-ray photons on a Ce:YAG scintillator. We demonstrate focusing of the soft-x-ray beam to a 1-microm spot size with a peak intensity of 1 x 10(14) W/cm2, which is to our knowledge the highest ever reported in the soft-x-ray region. PMID- 15357363 TI - Optimization of linearity by use of a glass plate in carousel interferometers. AB - Carousel interferometers are compact swinging interferometers, but their popularity is limited because of significant nonlinearity in rotational measurements. Introducing a glass plate in a rotating interferometer and optimizing its thickness has improved the linearity drastically. Computer simulations show reductions in nonlinearity of several orders of magnitude. PMID- 15357364 TI - All-optical display using photoinduced anisotropy in a bacteriorhodopsin film. AB - Photoinduced anisotropy in a bacteriorhodopsin film using the pump-probe method was investigated. A diode-pumped second-harmonic YAG laser was used as the pump beam, and three wavelengths, at lambda = 442, 532, 655 nm, from different lasers were used as probe beams. Without the pump beam, the probe light cannot transmit the analyzer to the detector. However, because of photoinduced anisotropy, a portion of the probe light is detected when the pump beam is present. Based on this property, we demonstrate a full-color all-optical display. PMID- 15357365 TI - Sub-shot-noise phase quadrature measurement of intense light beams. AB - We present a setup for performing sub-shot-noise measurements of the phase quadrature of intense pulsed light without the use of a separate local oscillator. A Mach-Zehnder interferometer with an unbalanced arm length is used to detect the fluctuations of the phase quadrature at a single sideband frequency. With this setup, the nonseparability of a pair of quadrature-entangled beams is demonstrated experimentally. PMID- 15357366 TI - Orbital angular momentum of entangled counterpropagating photons. AB - We elucidate the paraxial orbital angular momentum of entangled photon pairs generated by spontaneous parametric downconversion in different noncollinear geometries in which the entangled photons counterpropagate. We find, in particular, the orbital angular momentum of entangled pairs generated in transverse-emitting configurations, in which none of the known rules for selecting orbital angular momentum holds. PMID- 15357367 TI - Vortices in femtosecond laser fields. AB - We experimentally generate optical vortices in the output beam of a 20-fs Ti:sapphire laser. Screw phase dislocations are imposed on the spectral components of the short pulses by aligning a computer-generated hologram in a dispersionless 4f setup. PMID- 15357368 TI - Wide spectral range third-order autocorrelator based on ultrafast nonresonant nonlinear refraction. AB - We demonstrate a simple scheme for a wide spectral range, third-order autocorrelator based on ultrafast nonlinear Kerr-type refraction. The technique was successfully used to characterize high-energy ultrashort pulses at 1550 and 1300 nm, where the pulse's shape and width are two of the most critical parameters. Because of its simplicity, this technique is also a powerful tool for the optimization of high-power chirped-pulse amplified laser systems, in which slight misalignment of the stretcher-compressor gratings can lead to spatiotemporal pulse distortions. In addition, it can be extended to low-power mode-locked oscillators. PMID- 15357369 TI - The metabolic syndrome--early action to decrease risks for cardiovascular disease. PMID- 15357370 TI - Managing transitional work--program foundation. AB - A comprehensive TWP is a critical element in any disability management effort. The program must be developed just as any successful worksite program is: with management support, teamwork, a coordinator, written policies and procedures, effective marketing, and communications. In addition, the program must be evaluated and continuously improved. A successful program will contribute to the location's bottom line as well as improve the health of employees. PMID- 15357371 TI - Processes of care for individuals with work related asthma: treatment characteristics and impact of asthma on work. AB - The prevalence of asthma among working adults continues to rise each year. The California Department of Health Services conducts surveillance of work related asthma (WRA) to classify each work related exposure using Doctor's First Reports of Occupational Illness and Injury (DFRs). Using a cross-sectional, descriptive, comparative design, additional interviews were conducted and medical records were reviewed to explore workers' and providers' perceptions of follow up care. Two cohorts were compared: workers with WRA who belonged to a large, single HMO (n = 79) and workers with WRA who underwent follow up outside this HMO (n = 76). The interview asked about providers seen, tests ordered, and the impact of asthma on work. The HMO clients were significantly more likely than the non-HMO clients to see occupational medicine specialists (p = .004) and have pulmonary function testing (p = .049) during initial treatment. Twenty-four percent of clients currently working reported missed workdays caused by asthma in the past 6 months. The findings indicate management of WRA varies by health care system in California. PMID- 15357372 TI - Education of garment workers: prevention of work related musculoskeletal disorders. AB - This educational intervention was designed as part of a garment worker occupational health and safety initiative, with the goal to reduce musculoskeletal symptoms in this monolingual Cantonese speaking population. Using risk communication and the Chinese concepts of yin and yang, the class curriculum was designed to be participatory. It focused on linking symptoms to high risk work activities; explaining the nature of musculoskeletal injury; and encouraging compliance with self care measures of ice, stretching, and early symptom reporting. A total of 21 women completed the Healthy Work Classes, with an increase in perceived levels of energy measured after each class. Additionally, contingency contracting for both individual and workplace change was piloted. This curriculum was revised to become a "train the trainer" program, with training of garment worker leaders and the goal to disseminate this prevention based curriculum to garment workers in the Oakland, California community. PMID- 15357373 TI - Diabetes update: promoting effective disease management. AB - Diabetes is common, serious, costly, and controllable. Current scientific evidence indicates much of the morbidity and mortality associated with diabetes can be eliminated through prevention, early detection, improved delivery of care, and better education for diabetes self management. Unfortunately, a wide gap still exists between current and desired diabetes care and practices. Using existing tools (see Figure) (Kentucky Diabetes Network, 2004), the occupational health nurse can help bridge this gap for employees with diabetes by assuring a workplace that supports achievement of successful outcomes related to diabetes management and control. PMID- 15357374 TI - Expanding food portions contribute to overweight and obesity. PMID- 15357375 TI - BVA dismay at the Government's response to the EFRACom report. PMID- 15357376 TI - Field study of the prevalence of lameness in horses with back problems. AB - A population of 805 horses (70 per cent dressage, 20 per cent show jumpers and 10 per cent trotters) with orthopaedic problems was examined for signs of lameness and back problems, irrespective of their original complaints. In the horses with a back problem the prevalence of lameness was 74 per cent, and back problems were diagnosed in 32 per cent of the lame horses. These percentages were significantly higher than those recorded in a control population of 399 horses, of which 20 per cent were lame and 12 per cent had back problems. In the group of horses with orthopaedic problems there was a strong association between lameness and back problems and, in particular, there was a high prevalence of lameness among the horses with back problems. PMID- 15357377 TI - Digestive pathology of sea turtles stranded in the Canary Islands between 1993 and 2001. AB - Digestive lesions were observed in 84 of 136 sea turtles (128 Caretta caretta, four Chelonia mydas and four Dermochelys coriacea) stranded in the Canary Islands between January 1993 and December 2001. In the oral cavity ulcerative and necropurulent stomatitis were the most frequently observed lesions, and in the oesophagus ulcerative and fibrinous oesophagitis, and traumatic oesophageal perforation were most frequently observed; all these lesions were mainly associated with the ingestion of fishing hooks. Different histological types of gastritis were observed in 35 of the turtles; necropurulent and fibrinous gastritis were associated with bacterial infections caused mainly by Proteus species, Vibrio alginolyticus, and Staphylococcus species, and larval nematodes of the genus Anisakis were responsible for a form of parasitic gastritis observed in 16 of the turtles. Different histological types of enteritis, including catarrhal, fibrinous, necropurulent and necrotising enteritis, affected 36 turtles; a wide range of gram-negative and gram-positive bacteria, including Bacillus species, Escherichia coli, Pasteurella species, Proteus species, Staphylococcus species, Streptococcus species and V. alginolyticus, were isolated from these lesions. All the cases of necrotising enteritis were associated with intestinal intussusception caused by the ingestion of monofilament fishing lines. Necrotising and/or multifocal granulomatous hepatitis were the lesions most commonly observed in the liver; they affected 29 of the turtles and were associated with Aeromonas hydrophila, Citrobacter species, E. coli, Proteus species, Staphylococcus species and V. alginolyticus infections. According to the stranding reports and the gross and histological lesions observed, 33 of the turtles had digestive lesions associated with the ingestion of hooks and monofilament lines, and two had lesions associated with the ingestion of crude oil. PMID- 15357378 TI - Accidental poisoning of 17 dogs with lasalocid. AB - Over a period of 10 days, 17 dogs became weak and developed neurological deficits of different degrees of severity. About 12 hours before these clinical signs appeared they had all eaten a particular brand of commercial dog food from a recently opened bag. They were all quadriparetic and hyporeflexic, and some of them also showed additional systemic or neurological signs, including dyspnoea, a high body temperature, tongue laxity, hyperaesthesia and anisochoria. Serum biochemical abnormalities included high activities of creatine kinase, lactate dehydrogenase and aspartate aminotransferase. Analysis of the suspect food revealed high concentrations of the ionophore lasalocid. Fifteen of the dogs were given supportive treatment at home and two were hospitalised. Five of the dogs died, but the others improved gradually and had fully recovered by one to four days after the appearance of the clinical signs. PMID- 15357379 TI - Genetic variability of the PrP gene in a goat herd in the UK. PMID- 15357380 TI - Phylogenetic analysis of type 2 porcine circoviruses identified in wild boar in Slovenia. PMID- 15357381 TI - First report of Cheyletiella parasitovorax infestation in the Eurasian badger (Meles meles). PMID- 15357382 TI - Export/import of ferrets under the Pet Travel Scheme. PMID- 15357383 TI - Docking dogs' tails. PMID- 15357384 TI - Riding establishments and the Animal Welfare Bill. PMID- 15357385 TI - New rules on bovine semen production. PMID- 15357386 TI - Apical tooth root abscesses in horses. PMID- 15357387 TI - Effect of grazing by poultry on the numbers of sheep parasitic larvae found on organic grassland. PMID- 15357388 TI - Advances in the knowledge of amphizoic amoebae infecting fish. AB - Free-living amoebae infecting freshwater and marine fish include those described thus far as agents of fish diseases, associated with other disease conditions and isolated from organs of asymptomatic fish. This survey is based on information from the literature as well as on our own data on strains isolated from freshwater and marine fish. Evidence is provided for diverse fish-infecting amphizoic amoebae. Recent progress in the understanding of the biology of Neoparamoeba spp., agents responsible for significant direct losses in Atlantic salmon and turbot industry, is presented. Specific requirements of diagnostic procedures detecting amoebic infections in fish and taxonomic criteria available for generic and species determination of amphizoic amoebae are analysed. The limits of morphological and non-morphological approaches in species determination are exemplified by Neoparamoeba, Vannella and Platyamoeba spp., which are the most common amoebae isolated from fish gills, Acanthamoeba and Naegleria spp. isolated from various organs of freshwater fish, and by other unique fish isolates of the genera Nuclearia, Thecamoeba and Filamoeba. Advances in molecular characterisation of SSU rRNA genes and phylogenetic analyses based on their sequences are summarised. Attention is particularly given to specific diagnostic tools for fish-infecting amphizoic amoebae and ways for their further development. PMID- 15357389 TI - Haemogregarina bigemina (Protozoa: Apicomplexa: Adeleorina)--past, present and future. AB - This paper reviews past, current and likely future research on the fish haemogregarine, Haemogregarina bigemina Laveran et Mesnil, 1901. Recorded from 96 species of fishes, across 70 genera and 34 families, this broad distribution for H. bigemina is questioned. In its type hosts and other fishes, the parasite undergoes intraerythrocytic binary fission, finally forming mature paired gamonts. An intraleukocytic phase is also reported, but not from the type hosts. This paper asks whether stages from the white cell series are truly H. bigemina. A future aim should be to compare the molecular constitution of so-called H. bigemina from a number of locations to determine whether all represent the same species. The transmission of H. bigemina between fishes is also considered. Past studies show that young fish acquire the haemogregarine when close to metamorphosis, but vertical and faecal-oral transmission seem unlikely. Some fish haemogregarines are leech-transmitted, but where fish populations with H. bigemina have been studied, these annelids are largely absent. However, haematophagous larval gnathiid isopods occur on such fishes and may be readily eaten by them. Sequential squashes of gnathiids from fishes with H. bigemina have demonstrated development of the haemogregarine in these isopods. Examination of histological sections through gnathiids is now underway to determine the precise development sites of the haemogregarine, particularly whether merozoites finally invade the salivary glands. To assist in this procedure and to clarify the internal anatomy of gnathiids, 3D visualisation of stacked, serial histological sections is being undertaken. Biological transmission experiments should follow these processes. PMID- 15357390 TI - The capsalidae (Monogenea: Monopisthocotylea): a review of diversity, classification and phylogeny with a note about species complexes. AB - The Capsalidae are monogeneans parasitizing 'skin', fins and gills of marine fishes. Some capsalids are pathogenic to cultivated fish and a few have caused epizootic events. It is a cosmopolitan family with broad host associations (elasmobranchs and teleosts, including sturgeons). Approximately 200 capsalid species are placed in nine subfamilies and 44-46 genera, some of which are well known (Benedenia, Capsala, Entobdella, Neobenedenia). Sturgeons host two capsalid species (Nitzschiinae) and 15 species in five genera are reliably reported from elasmobranchs. The combination of ancient (shark, ray, sturgeon) and modem (teleost) host fish lineages indicates that capsalid evolution is likely a blend of coevolution and host-switching, but a family phylogeny has been lacking due to deficient knowledge about homologies. The current phenetic subfamilial classification is discussed in detail using a preliminary phylogeny generated from large subunit ribosomal DNA sequence data from representatives of five subfamilies. Monophyly of the Capsalidae is supported by possession of accessory sclerites. Hypotheses are proposed for the possible radiation of capsalids. A suggestion that Neobenedenia melleni, a pathogenic species atypical due to its broad host-specificity (>100 host teleost species from >30 families in five orders), may be a complex of species is supported from genetic evidence. This may explain peculiarities in biology, taxonomy, host associations and geographic distribution of N. 'melleni' and has implications for fish health. Holistic studies using live and preserved larval and adult capsalid specimens and material for genetic analysis are emphasised to further determine identity, phylogeny and details of biology, especially for pathogenic species. PMID- 15357391 TI - A review of Dendromonocotyle (Monogenea: Monocotylidae) from the skin of stingrays and their control in public aquaria. AB - Dendromonocotyle species (Monogenea: Monocotylidae) are the only monocotylids to parasitize the skin of chondrichthyan hosts. Currently 11 species are recorded from the skin of ray species in the Dasyatidae, Myliobatidae and Urolophidae. There have been increasing reports of Dendromonocotyle outbreaks on rays kept in public aquaria. This paper provides a broad review of Dendromonocotyle that should assist taxonomists and aquarists with species identification and help decisions on potential control methods for Dendromonocotyle infections. The taxonomy and host-specificity of Dendromonocotyle are discussed and a key to current species is provided. We summarise what little is known about the biology of Dendromonocotyle including egg embryonation and hatching, feeding, camouflage and reproduction. The efficacy of freshwater baths, chemical treatments and biological control measures such as the use of cleaner fish for Dendromonocotyle is also discussed. We demonstrate that effective control of Dendromonocotyle on captive rays is hampered by the lack of basic biological data on the life cycle of the parasites. A case history is provided outlining the success of a public aquarium (Underwater World, Mooloolaba, Queensland, Australia) in managing D. pipinna infections on captive Taeniura meyeni without chemical intervention simply by taking measures to reduce host stress. PMID- 15357392 TI - Larvae of gryporhynchid cestodes (Cyclophyllidea) from fish: a review. AB - Larvae (metacestodes) of tapeworms of the cyclophyllidean family Gryporhynchidae (previously included in the Dilepididae) occur in different internal organs of fresh- and brackish water fish (110 fish species of 27 families in 12 orders reported), which serve as the second intermediate hosts. The species composition, spectrum of fish hosts, sites of infection, and geographical distribution of gryporhynchids recorded from fish are reviewed here on the basis of literary data and examination of extensive material from helminthological collections. Metacestodes of the following genera have been found in fish: Amirthalingamia Bray, 1974 (1 species), Ascodilepis Guildal, 1960 (1), Cyclustera Fuhrmann, 1901 (4), Dendrouterina Fuhrmann, 1912 (1), Glossocercus Chandler, 1935 (3), Neogryporhynchus Baer et Bona, 1960 (1), Paradilepis Hsu, 1935 (5), Parvitaenia Burt, 1940 (2), and Valipora Linton, 1927 (3). However, most published records concern only three species, namely Neogryporhynchus cheilancristrotus (Wedl, 1855) from the intestinal lumen, Paradilepis scolecina (Rudolphi, 1819) from the liver and mesenteries, and Valipora campylancristrota (Wedl, 1855) from the gall bladder of cyprinids and other fish in the Palaearctic Region. Data on other species as well as reports from other regions are very scarce and almost no information is available from Australia, tropical Asia and South America. A recent study of gryporhynchid metacestodes from Mexico (Scholz and Salgado Maldonado 2001), which reported 13 species, suggested that they may be more common than indicated by records in the literature. Although only a few cases of pathogenic influence of larvae on fish hosts have been reported, the veterinary importance of gryporhynchids remains to be assessed on the basis of more detailed studies. The data available indicate a strict host and site specificity of some species whereas others occur in a wide spectrum of fish hosts and are not strictly site-specific. Evaluation of Paradilepis larvae from the liver of salmonid fish from British Columbia, Canada, identified as P. simoni Rausch, 1949 by Ching (1982), has shown that they probably belong to two species, P. simoni and P. rugovaginosus Freeman, 1954. Metacestodes of the latter species and those of Cyclustera magna (Baer, 1959) from the intestinal wall of Tilapia zillii (Gervais) from Kenya are reported from fish for the first time. PMID- 15357393 TI - Defeating diplostomoid dangers in USA catfish aquaculture. AB - Diplostomoid digenean metacercariae have caused widescale mortalities of channel catfish, Ictalurus punctatus (Rafinesque), at aquaculture farms in Louisiana, Mississippi and Arkansas, USA. Originally, based on a tentative diagnosis, the industry considered the primary harmful agent to be an introduced species from Europe, Bolbophorus confusus (Krause, 1914), frequently reported from the American white pelican, Pelecanus erythrorhynchos Gmelin. Our group has now shown, using ITS 1-2 plus three more-conservative gene fragments, that two sympatric species of Bolbophorus exist in the American white pelican. One, B. damnificus Overstreet et Curran, 2002, infects the musculature of catfish, and the other, probably not B. confusus, does not infect catfish. However, at least four other pathogenic diplostomoids and a clinostomoid infect the catfish, and they use at least four different snail hosts, including the planorbids Planorbella trivolvis (Say) and Gyraulus parvus (Say), the physid Physella gyrina (Say) and a lymnaeid. Two metacercariae, B. damnificus and Bursacetabulus pelecanus Dronen, Tehrany et Wardle, 1999, infect the catfish and mature in the pelican; two others, Austrodiplostomum compactum (Lutz, 1928) and Hysteromorpha cf. triloba (Rudolphi, 1819), mature in cormorants; one, Diplostomum sp., matures in seagulls and at least one, Clinostomum marginatum (Rudolphi, 1819), matures in herons, egrets and other wading birds. Consequently, management of catfish ponds relative to digenean infections requires considerable biological information on the fish, bird, and snail hosts as well as the parasites. PMID- 15357394 TI - A microsporidian parasite of the genus Spraguea in the nervous tissues of the Japanese anglerfish Lophius litulon. AB - In the present study, a high percentage of Japanese anglerfish, Lophius litulon (Jordan, 1902), contained a microsporidian infection of the nervous tissues. Xenomas were removed and prepared for standard wax histology and transmission electron microscopy (TEM). DNA extractions were performed on parasite spores and used in PCR and sequencing reactions. Fresh spores measured 3.4 x 1.8 microm and were uniform in size with no dimorphism observed. TEM confirmed that only a single developmental cycle and a single spore form were present. Small subunit (SSU) rDNA sequences were >99.5% similar to those of Spraguea lophii (Doflein, 1898) and Glugea americanus (Takvorian et Cali, 1986) from the European and American Lophius spp. respectively. The microsporidian from the nervous tissue of L. litulon undoubtedly belongs in the genus Spraguea Sprague et Vivra, 1976 and the authors suggest a revision to the generic description of Spraguea to include monomorphic forms and the transfer of Glugea americanus to Spraguea americana comb. n. Since no major differences in ultrastructure or SSU rDNA sequence data exist between Spraguea americana and the microsporidian from the Japanese anglerfish, they evidently belong to the same species. This report of Spraguea americana is the first report of a Spraguea species from L. litulon and indeed from the Pacific water mass. PMID- 15357395 TI - Histophagous scuticociliatids (Ciliophora) parasitizing turbot Scophthalmus maximus: morphology, in vitro culture and virulence. AB - Systemic ciliatosis caused by histophagous ciliates constitutes a serious disease of cultured turbot. Six ciliate isolates were obtained from parasitized turbot during six epizootics at four different farms located in Spain, France and Portugal. Axenic cultures of the six isolates were obtained by periodical subculturing in ATCC 1651MA or supplemented L-15 media. In basal media or seawater, the parasites could survive starving for long periods with no apparent proliferation. In adequate media, growth kinetics was found to be very similar for isolates A and B, with a clear influence of temperature. Morphological studies demonstrated that all isolates share common features that allows their assignment to either Philasterides Kahl, 1931 or Miamiensis Thompson et Moewus, 1964. However, statistically significant differences were evident in pairwise comparisons of the isolates from the four farm sites in 16 taxonomically relevant morphometric features. This could allow the discrimination of different species or strains. Virulence of isolates A and B for healthy turbot was tested in several experiments. Differences in the virulence were especially evident after long-term in vitro culturing, isolate A being clearly attenuated after 35-42 passages, whereas isolate B became more virulent after 20-42 passages. The need of further studies to confirm such virulence variability and its implications in pathogenesis and prevention of turbot scuticociliatoses is stressed. PMID- 15357396 TI - Studies on transmission and life cycle of Enteromyxum scophthalmi (Myxozoa), an enteric parasite of turbot Scophthalmus maximus. AB - In order to elucidate the transmission and dispersion routes used by the myxozoan parasite Enteromyxum scophthalmi Palenzuela, Redondo et Alvarez-Pellitero, 2002 within its host (Scophihalmus maximus L.), a detailed study of the course of natural and experimental infections was carried out. Purified stages obtained from infected fish were also used in in vitro assays with explants of uninfected intestinal epithelium. The parasites can contact and penetrate loci in the intestinal epithelium very quickly. From there, they proliferate and spread to the rest of the digestive system, generally in an antero-posterior pattern. The dispersion routes include both the detachment of epithelium containing proliferative stages to the intestinal lumen and the breaching of the subepithelial connective system and local capillary networks. The former mechanism is also responsible for the release of viable proliferative stages to the water, where they can reach new fish hosts. The finding of parasite stages in blood smears, haematopoietic organs, muscular tissue, heart and, less frequently, skin and gills, suggests the existence of additional infection routes in transmission, especially in spontaneous infections, and indicates the role of vascular system in parasite dispersion within the fish. The very high virulence of this species in turbot and the rare development of mature spores in this fish may suggest it is an accidental host for this parasite. This may also question the existence of a two-host life cycle involving an actinosporean stage in this species. Further studies are needed to clarify this open point of the life cycle. PMID- 15357397 TI - Intraoligochaete development of Myxobolus intimus (Myxosporea: Myxobolidae), a gill myxosporean of the roach (Rutilus rutilus). AB - The infection with Myxobolus intimus Zaika, 1965 in the gills of the roach Rutilus rutilus (L.) from Lake Balaton was recorded in 28 out of the 39 fish examined. Developing and mature plasmodia were detected on the gills exclusively in the spring. The Myxobolus intimus infection was found only in 2- to 3-year-old fish. In histological sections, young plasmodia were found in capillaries of the secondary lamellae. More mature, round plasmodia 0.4-0.6 mm in diameter, deformed the respiratory lamellae. The intraoligochaete development of M. intimus was studied in experimentally infected oligochaetes. In two experiments, uninfected Tubifex tubifex Muller and Limnodrilus hoffmeisteri (Claparede) were exposed to mature myxospores of M. intimus. In both experiments, typical triactinospores developed in T. tubifex specimens but no infection was found in L. hoffmeisteri. In semithin sections, developmental stages, pansporocysts and actinospores, were found within the proliferated gut epithelium of T. tubifex. Triactinospores were first released from oligochaetes 37 and 58 days after initial exposure in the two experiments, respectively. Each triactinospore had three pyriform polar capsules and a cylindrical sporoplasm with 32 secondary cells. The spore body joined the 3 caudal projections with a moderately long style. PMID- 15357398 TI - Sphaerospora dicentrarchi and S. testicularis (Myxozoa: Sphaerosporidae) in farmed European seabass (Dicentrarchus labrax) from Italy. AB - The distribution of Sphaerospora dicentrarchi Sitja-Bobadilla et Alvarez Pellitero, 1992 and S. testicularis Sitja-Bobadilla et Alvarez-Pellitero, 1990, myxozoan parasites of European seabass Dicentrarchus labrax (L.), was investigated in different farming systems in Italy. In total, 1406 fish were examined. High S. dicentrarchi prevalence was observed in all the farming systems involved in this survey (extensive farms: 51.5%; intensive farms: inland 59.6%, inshore floating cages 76.2%, offshore floating cages 41.6%) except for submersible cages (7.4%). S. testicularis was detected only in nine male fish from two intensive farms. The epidemiology and pathological effects of the parasites are discussed. PMID- 15357399 TI - The phylogeny of marine and freshwater species of the genus Chloromyxum Mingazzini, 1890 (Myxosporea: Bivalvulida) based on small subunit ribosomal RNA gene sequences. AB - The small subunit ribosomal RNA gene (SSU rDNA) of two freshwater and one marine species of the genus Chloromyxum Mingazzini, 1890 were sequenced. The SSU rDNA trees obtained show the phylogenetic position of the marine species Chloromyxum leydigi Mingazzini, 1890 to be at the base of the freshwater clade, being well supported by a high bootstrap value. Chloromyxum cyprini Fujita, 1927 is closely related to Chloromyxum truttae Leger, 1906 and they represent a sister branch to raabeia sp., Myxidium sp. and Myxidium truttae Leger, 1930. Chloromyxum legeri Tourraine, 1931 is in a position ancestral to Myxidium lieberkuehni Butschli, 1882 and Sphaerospora oncorhynchi Kent, Whitaker et Margolis, 1993. Three newly sequenced species of the genus Chloromyxum represent three separate lineages within the myxosporean tree and do not support the monophyly of this genus. PMID- 15357400 TI - Extensive release of an antigen associated with the sporogonic stages of myxobolus cerebralis (Myxozoa: Myxosporea) is detected by a heterologous antibody raised to Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea). AB - Monoclonal antibody B4 (mAb B4) was previously developed to the myxozoan parasite Tetracapsuloides bryosalmonae Canning, Curry, Feist, Longshaw et Okamura, 1999, the causative agent of proliferative kidney disease of salmonids, Here we describe the reaction of mAb B4 against Myxobolus cerebralis Hofer, 1903, the parasite that causes 'whirling disease' in salmonids. Tissues examined were collected from experimentally infected rainbow trout Oncorhynchus mykiss (Walbaum) and the aquatic oligochaete Tubifex tubifex (O.F. Muller), the two hosts involved in the life cycle of M. cerebralis. Paraffin sections of infected rainbow trout taken at 4 h and 3, 10, 17 and 54 days post-exposure to infective M. cerebralis actinospores were immunohistochemically stained with mAb B4. Longitudinal sections through infected T. tubifex sampled 120 days post-exposure to M. cerebralis myxospores were also examined using this method. The only phase of the M. cerebralis life cycle that expressed the mAb B4 antigen was during sporogenesis in the salmonid host. The immunohistochemical staining demonstrated that the antigen was released into the tissues surrounding the spore and sporogonic stages of the parasite. The localisation of the antigen was diffuse in the fish, suggesting that the possible effect of M. cerebralis infection is extensive through the head tissues and not limited to areas of cartilage destruction as previously thought. PMID- 15357401 TI - Proliferative renal myxosporidiosis in spawning coho salmon (Oncorhynchus kisutch) in British Columbia and Washington. AB - An unidentified myxosporean parasite (CKX) is described from the kidney of approximately 80% of spawning coho salmon Oncorhynchus kisutch (Walbaum) in British Columbia, Canada and Washington, United States of America. Morphological features were described using light and electron microscopy. Sequencing of polymerase chain reaction (PCR) amplified 18S ribosomal RNA gene and in situ hybridisation were used to further characterise CKX. The parasite occurred with a focal distribution within tubule epithelial cells, the tubule lumen and the interstitium as primary cells containing from one to at least 16 secondary cells. Luminal stages were degenerate and sporogony was not observed. In situ hybridisation using a digoxygenin-labelled DNA probe confirmed CKX to be the source of DNA used in PCR analyses. CKX 18S rDNA sequences were most similar (97%) to those of Sphaerospora oncorhynchi. Phylogenetic analysis revealed similarities among the 18S rDNA sequences of CKX, S. oncorhynchi and Myxidium lieberkuehni. CKX is hypothesised to be the abortive extrasporogonic development of a Sphaerospora sp. or Myxidium sp. occurring in immune-incompetent spawning and senescent salmon. PMID- 15357402 TI - Some aspects of the biology and life history of Bothitrema bothi (Monogenea: Bothitrematidae) from the flounder Scophthalmus aquosus (Bothidae) from New Jersey, USA. AB - Bothitrema bothi (MacCallum, 1913), a monopisthocotylean monogenean (Bothitrematidae) found on gills and occasionally on the olfactory rosette within the nares of the bothid flounder, Scophthalmus aquosus (Mitchill, 1815), is distributed along the Atlantic coast of the USA from Woods Hole, Massachusetts to Delaware Bay. This study details aspects of the morphology, microecology and biology of the oncomiracidium, juvenile and adult. Adult worms occur almost exclusively between rakers on the first and second gill arches while juveniles occupy secondary gill filaments. Analysis of variance revealed significant arch, side and position effects. Worm attachment and position is maintained largely by suction generated by a circular seal using its 54 radially arranged accessory haptoral sclerotised elements and a marginal valve. A small anterior appendix with two marginal hooks lies dorsal to a deep anteromedian cleft in the haptor, which, together with wall muscles, creates and releases the negative pressure required for attachment. Four pairs of gland openings in the ventrolateral margins of the anterior adhesive area produce secretions for attachment. Operculate eggs with four rounded protuberances and a filament are laid individually at four-minute intervals at 22 degrees C. Hatching occurs in five days at 20 degrees C. Oncomiracidia have two pairs of eyes, with the posterior pair fused, show both positive and negative phototaxis and swim at a mean rate of 5.2 mm/s at 22 degrees C. Larvae have a circle of 16 hooks and a pair of hamulus primordia. Other accessory haptoral sclerotised structures arise de novo in juveniles after attachment to the host. These data will help to resolve more clearly the relationships of the Bothitrematidae within the Monopisthocotylea. PMID- 15357403 TI - The use of morphometric characters to discriminate specimens of laboratory-reared and wild populations of Gyrodactylus salaris and G. thymalli (Monogenea). AB - Gyrodactylus thymalli Zitnan, 1960 and G. salaris Malmberg, 1957 have an indistinguishable ribosomal internal transcribed spacer (ITS) DNA sequence, but exhibit surprisingly high levels of intra- and interspecific sequence variation of the mitochondrial cytochrome oxidase I (CO1) gene. To test whether different populations of these reportedly very similar species could be discriminated using morphometric methods, we examined the morphometry of four different populations representing different mitochondrial clades. Twenty five point-to-point measurements, including five new characters of the attachment hooks, were recorded from three Norwegian laboratory populations (G. salaris from the Rivers Lierelva and Rauma, and G. thymalli from the River Rena), and from one wild population of G. thymalli from the River Test, UK. The Norwegian populations were kept under identical environmental conditions to control for the influence of temperature on the haptoral attachment hooks. Data were subsequently subjected to univariate and linear stepwise discriminant analyses. The model generated by the linear stepwise discriminant analysis used 18 of the 25 original variables, the first two roots accounting for 96.6% of the total variation between specimens. The hamulus shaft length accounts for 66.7% of the overall correct classification efficiency. Based on morphometry, all specimens were assigned to the correct species. Apart from three specimens of G. salaris from the River Lierelva population which were misclassified as belonging to the G. salaris Rauma population, all specimens were assigned to the correct population. Thus, populations of Gyrodactylus identified by mtDNA can also be discriminated using morphometric landmark distances. PMID- 15357404 TI - Chaetotaxy applied to Norwegian Gyrodactylus salaris Malmberg, 1957 (Monogenea) clades and related species from salmonids. AB - Gyrodactylus salaris Malmberg, 1957 is a major pathogen of wild Salmo salar L. parr populations in Norway, and its delimitation from non-pathogenic species is important. The present study was undertaken to test the power of chaetotaxy to differentiate between three populations belonging to both the same and different clades (as stated by mtDNA) of G. salaris, in addition to three different species of gyrodactylids (G. salaris, G. thymalli and G. caledoniensis). The gyrodactylids were processed for chaetotaxy in situ and a maximum of 50 specimens per collection site were used to construct a generalised map over the sensilla. The sensilla were found in all populations to be symmetrically distributed around the median longitudinal axis, according to a formula of 7 dorsal (34 sensilla) and 8 ventral (44 sensilla) clusters on each side of the median line. The three Norwegian populations of G. salaris were found identical, as were the population of G. thymalli. The specimens of G. caledoniensis from Scotland, however, were found to differ from the Norwegian species G. salaris and G. thymalli by the position of one sensillum in two of the clusters. A comparison of the sensillum pattern of laboratory maintained G. salaris (River Lierelva) with results obtained ten years earlier, questions the temporal stability of the chaetotaxy pattern. The present results indicate that chaetotaxy can be used to discriminate between certain Gyrodactylus spp. but not generally. PMID- 15357405 TI - Homing of Gyrodactylus salaris and G. derjavini (Monogenea) on different hosts and response post-attachment. AB - In natural European waters, the congeneric monogeneans Gyrodactylus derjavini Mikailov, 1975 and G. salaris Malmberg, 1957 are primarily found on brown trout Salmo trutta L. and Atlantic salmon Salmo salar L., respectively. Interestingly, rainbow trout, Oncorhynchus mykiss (Walbaum), originating from North America, is as susceptible as brown trout to G. derjavini. However, the mechanisms involved in this host specificity are poorly understood but may include behavioural, mechanical and chemical factors affecting parasite attraction, attachment, feeding, reproduction and host responses. In the present laboratory work, this question has been studied. Detached parasites (either G. derjavini or G. salaris) were offered a choice in small aquaria between fry of rainbow trout, Atlantic salmon and carp Cyprinus carpio L. Within 48 hours more than 90% of G. derjavini colonised rainbow trout and left salmon almost uninfected. Some parasites were found on carp. During the same time span, more than 60% of G. salaris attached to salmon, the rest infected rainbow trout and none were found on carp. Following attachment, the parasites need appropriate stimuli to initiate feeding and reproduction but even such a successful specific colonisation can be followed by a host response. Both humoral and cellular elements have been suggested to participate in these reactions but in the present work it was demonstrated by immunoblotting and immunocytochemistry that no antibodies in host mucus and host plasma bound to any parasite structures or epitopes. PMID- 15357406 TI - The bathymetric distribution of the digenean parasites of deep-sea fishes. AB - The bathymetric range of 149 digenean species recorded deeper than 200 m, the approximate depth of the continental shelf/slope break, are presented in graphical form. It is found that only representatives of the four families Lepocreadiidae, Fellodistomidae, Derogenidae and Hemiuridae reach to abyssal regions (>4000 m). Three other families, the Lecithasteridae, Zoogonidae and Opecoelidae, have truly deep-water forms reaching deeper than 3000 m. Bathymetric data are available for the Acanthocolpidae, Accacoeliidae, Bucephalidae, Cryptogonimidae, Faustulidae, Gorgoderidae, Monorchiidae and Sanguinicolidae showing that they reach deeper than 200 m. No bathymetric data are available for the members of the Bivesiculidae and Hirudinellidae which are reported from deep sea hosts. These results indicate that only seventeen out of the 150 or so digenean families are reported in the deep sea. PMID- 15357407 TI - Tinnitus and cochlear implantation. AB - Tinnitus is a common experience, but there is very marked heterogeneity of aetiology, perception and the extent of distress among individuals who experience tinnitus. In view of this, a modern approach to tinnitus should consider homogeneous groups of individuals. This review considers tinnitus experiences in patients undergoing cochlear implantation, this being of interest because the prevalence of tinnitus in this patient group prior to surgery may shed some light on the link between cochlear dysfunction and tinnitus Second, any change in tinnitus experience as a result of electrode placement surgery or cochlear implant activation has relevance for patient counselling and informed consent. Finally, in recent tinnitus retraining therapy literature there has been the suggestion that unilateral sound therapy for tinnitus patients may set up an unhelpful asymmetry of input to the auditory system, with possible exacerbation of contralateral tinnitus. Unilateral cochlear implant use represents the most dramatic asymmetry possible and hence is a test of that hypothesis. Relevant papers (n = 32) were identified from literature databases. The standard of reporting tinnitus results was inconsistent. Tinnitus is experienced by up to 86% of adult cochlear implant candidates, but is not universal and is only troublesome in a small proportion (reported as 27% in one study). Electrode insertion may induce tinnitus in a small (up to 4%) number of patients, but this is rare. Cochlear implant device use is associated with reduction of tinnitus intensity and awareness in up to 86% of patients, and rarely with exacerbation (up to 9%). There are some indications in the literature that the more complex the simulation strategy, the larger that effect. Specifically, unilateral cochlear implant use was generally associated with reduction of contralateral tinnitus (in up to 67% of individuals) rather than exacerbation, and so the assertion that unilateral sound therapy for tinnitus is contraindicated is not proven. PMID- 15357408 TI - Longitudinal behaviour of neural response telemetry (NRT) data and clinical implications. AB - Neural response telemetry (NRT) data from 63 subjects equipped with the Nucleus CI24M Cochlear Implant System generally exhibited little change over up to 4 years. Larger changes, when they occurred, were seen only within the first 15 months postoperatively, and these changes diminished over time. Intraoperative NRT data were generally stable enough to be used for assisting in the initial speech processor fitting sessions. It was not possible to predict changes in the subjective map threshold and comfortable loudness levels (T and C levels, respectively) based on observed changes in the NRT data. The long-term stability of the neural response amplitude and the neural response threshold, however, implies that NRT may be useful as a routine diagnostic tool to detect changes to the neural periphery over time. PMID- 15357409 TI - Effects of amplitude compression on first- and second-order modulation detection thresholds in cochlear implant listeners. AB - The aim of this study was to examine the effects of instantaneous non-linear amplitude mapping on the detection of single-component and multicomponent temporal envelopes. To address this issue, first- and second-order amplitude modulation detection thresholds were measured in four cochlear implant users with the intervention of the compression device of the implant processor. The compression device is set to produce either a strongly or a weakly logarithmic mapping of stimulus amplitude to electrical amplitude. 'First-order' modulation detection thresholds indicate the ability of listeners to detect sinusoidal amplitude modulation (SAM) applied to a white noise carrier; they are measured as a function of the rate of that modulation, fm. 'Second-order' modulation detection thresholds indicate the ability to detect sinusoidal modulation applied to the depth of a sinusoidally amplitude-modulated signal (here, a 16-Hz sinusoidally amplitude-modulated white noise); they are measured as a function of the rate of the modulation applied to the modulation depth (referred to as fm'). In each task, stimuli are transformed by the implant processor and are presented through one electrode at approximately the same level. The results show that, in cochlear implant listeners, both first- and second-order modulation detection thresholds measured at the lower rates (< or =7 Hz) decrease slightly by about 3 6dB when the stronger compression is used. No effect of compression is observed at higher rates. These results suggest that instantaneous logarithmic amplitude mapping has beneficial- but limited-effects on the detection of single-component and multicomponent temporal envelopes. These results are discussed in light of current models of temporal envelope processing. PMID- 15357410 TI - Estimated variability of real-ear insertion response (REIR) due to loudspeaker type and placement. AB - The real-ear insertion response (REIR) of a hearing aid is estimated as the difference between the aided response and the unaided response in the ear canal. Changes in the position of the loudspeaker relative to the head, between the two measurements, may reduce the accuracy of the estimate. The spatial variability of the sound field at distances close to the loudspeaker is less for a 'flat-panel' loudspeaker than for a conventional cone loudspeaker; the panel might thus lead to a reduced influence of loud-speaker position. To assess this, we measured the real-ear unaided response (REUR) as each of three loudspeakers (two cone type and one panel) was moved in a three-dimensional space centred at either 0 degrees or 45 degrees azimuth, at a distance of 50cm from a KEMAR manikin. Contrary to our expectation, the variability of the REUR was larger for the panel than for the cone loudspeakers The REUR varied less with position for the 0 degrees than for the 45 degrees azimuth. The variability of the REUR decreased with increasing distance of the loudspeaker from KEMAR. We tentatively suggest that loudspeaker to-client distances of 40-50 cm should be used and that a 0 degrees azimuth is preferable. PMID- 15357411 TI - The origin of order effects in the results of the bi-thermal caloric test. AB - An order effect, characterized by a progressive decline in response magnitude, has been observed in the caloric test, and it has been suggested that the effect may be countered by applying a correction. This study was undertaken to determine whether the effect has its origin in physiological adaptation or in calibration drift of the recording system. Thirty-two normal subjects underwent the caloric test, in which their nystagmus was monitored simultaneously by both electronystagmography (ENG) and videonystagmography (VNG). The experimental design also allowed other aspects of the caloric test sequence to be investigated. No evidence of physiological adaptation of the response was seen. However, there was a significant change in corneoretinal potential, and hence ENG calibration, during the tests. This was more marked if subjects' eyes were maintained in darkness. It is recommended that no correction be made for order effects but that the calibration of ENG-based systems is adjusted immediately prior to each caloric irrigation. PMID- 15357412 TI - The effect of presentation level and compression characteristics on sentence recognition in modulated noise. AB - The effect of fast-acting compression on speech recognition in fully modulated (FUM) noise in listeners with normal and impaired hearing was investigated in two experiments We wanted to determine the relationships between the benefit from compression and some audiological factors. Furthermore, the sensitivity to changes in compression parameters was also evaluated. The results showed that two thirds of the listeners performed worse with fast-acting compression than with linear processing. Normal-hearing listeners showed the most benefit from compression. A significant relationship was found between benefit from compression and speech-to-noise ratio at threshold (SNRT) in slightly modulated (SM) noise. Pure-tone threshold was found to be a weak predictor of benefit from compression. No relationship was found between benefit from compression and the release of masking for the FUM noise. The variability in the results across different compression parameters was related to SNRT in SM noise. The results suggest an inverse relationship between benefit from compression and the severity of the suprathreshold hearing loss PMID- 15357413 TI - Presbyacusis-hearing thresholds and the ISO 7029. AB - The purpose of this study was to analyse the changes in pure-tone hearing thresholds with age. We studied a random sample of 232 elderly subjects with a battery of audiological tests, including pure-tone audiometry in the conventional and extended high-frequency range, using the normative distributions from the International Organization for Standardization (ISO) standard 7029 for comparison. Sixty otologically normal (ON) subjects were selected for comparative analysis with the unscreened (US) sample, and for description of gender and age group differences. With the use of a mathematical transformation of threshold data, it was found that the ISO 7029 normative alpha coefficient in females may be set too low compared to our sample in the lower frequencies, leading to an underestimation of hearing thresholds in ON females. In our ON sample, hearing thresholds deteriorated with age in the extended high-frequency audiometric range. No gender threshold differences were found, although the prevalence of unmeasurable responses was higher in males at some of these frequencies. The ON screening criteria in ISO 7029 may be unreliable in subjects over 60 years of age, as threshold differences between ON and US subjects were not consistent at any frequency. PMID- 15357414 TI - Low endolymph calcium concentrations in deafwaddler2J mice suggest that PMCA2 contributes to endolymph calcium maintenance. AB - In vertebrates, transduction of sound into an electrochemical signal is carried out by hair cells that rely on calcium to perform specialized functions. The apical surfaces of hair cells are surrounded by endolymphatic fluid containing calcium at concentrations that must be maintained by active transport. The mechanism of this transport is unknown, but an ATP-dependent pump is believed to participate. Mutation of the Atp2b2 gene that encodes plasma membrane calcium ATPase type 2 (PMCA2) produces the deaf, ataxic mouse: deafwaddler2J (dfw2J). We hypothesized that PMCA2 might transport calcium into the endolymph and that dfw2J mice would have low endolymph calcium concentrations, possibly contributing to their deafness and ataxia. First, using immunocytochemistry, we demonstrated that PMCA2 is present in control mice inner and outer hair cell stereocilia where it could pump calcium into the endolymph and that PMCA2 is absent in dfw2J stereocilia. Second, using an aspirating microelectrode and calcium-sensitive fluorescent dye, we found that dfw2J mice endolymph calcium concentrations are significantly lower than those of control mice. These findings suggest that PMCA2, located in hair cell stereocilia, contributes significantly to endolymph calcium maintenance. PMID- 15357415 TI - Postsynaptic targets of type II auditory nerve fibers in the cochlear nucleus. AB - Type II auditory nerve fibers, which provide the primary afferent innervation of outer hair cells of the cochlea, project thin fibers centrally and form synapses in the cochlear nucleus. We investigated the postsynaptic targets of these synapses, which are unknown. Using serial-section electron microscopy of fibers labeled with horseradish peroxidase, we examined the border of the granule-cell lamina in mice, an area of type II termination that receives branches having swellings with complex shapes. About 70% of the swellings examined with the electron microscope formed morphological synapses, which is a much higher value than found in previous studies of type II swellings in other parts of the cochlear nucleus. The high percentage of synapses enabled a number of postsynaptic targets to be identified. Most of the targets were small dendrites. Two of these dendrites were traced to their somata of origin, which were cochlear nucleus "small cells" situated at the border of the granule-cell lamina. These cells did not appear to receive any terminals containing synaptic vesicles that were large and round, indicating a lack of input from type I auditory nerve fibers. Nor did type II swellings or targets participate in the synaptic glomeruli formed by mossy terminals and the dendrites of granule cells. Other type II synapses were axosomatic and their targets were large cells, which were presumed multipolar cells and one cell with characteristics of a globular bushy cell. These large cells almost certainly receive additional input from type I auditory nerve fibers, which provide the afferent innervation of the cochlear inner hair cells. A few type II postsynaptic targets-the two small cells as well as a large dendrite-received synapses that had accompanying postsynaptic bodies, a likely marker for synapses of medial olivocochlear branches. These targets thus probably receive convergent input from type II fibers and medial olivocochlear branches. The diverse nature of the type II targets and the examples of segregated convergence of other inputs illustrates the synaptic complexity of type II input to the cochlear nucleus. PMID- 15357416 TI - Efferent actions in the chinchilla vestibular labyrinth. AB - Efferent fibers were electrically stimulated in the brain stem, while afferent activity was recorded from the superior vestibular nerve in barbiturate anesthetized chinchillas. We concentrated on canal afferents, but otolith afferents were also studied. Among canal fibers, calyx afferents were recognized by their irregular discharge and low rotational gains. In separate experiments, stimulating electrodes were placed in the efferent cell groups ipsilateral or contralateral to the recording electrode or in the midline. While single shocks were ineffective, repetitive shock trains invariably led to increases in afferent discharge rate. Such excitatory responses consisted of fast and slow components. Fast components were large only at high shock frequencies (200-333/s), built up with exponential time constants <0.1 s, and showed response declines or adaptation during shock trains >1 s in duration. Slow responses were obtained even at shock rates of 50/s, built up and decayed with time constants of 15-30 s, and could show little adaptation. The more regular the discharge, the larger was the efferent response of an afferent fiber. Response magnitude was proportional to cv*b, a normalized coefficient of interspike-interval variation (cv*) raised to the power b = 0.7. The value of the exponent b did not depend on unit type (calyx vs. bouton plus dimorphic, canal vs. otolith) or on stimulation site (ipsilateral, contralateral, or midline). Responses were slightly smaller with contralateral or midline stimulation than with ipsilateral stimulation, and they were smaller for otolith, as compared to canal, fibers. An anatomical study had suggested that responses to contralateral afferent stimulation should be small or nonexistent in irregular canal fibers. The suggestion was not confirmed in this study. Contralateral responses, including the large responses typically seen in irregular fibers, were abolished by shallow midline incisions that should have severed crossing efferent axons. PMID- 15357417 TI - Gentamicin pharmacokinetics in the chicken inner ear. AB - Avians have the unique ability to regenerate cochlear hair cells that are lost due to ototoxins or excessive noise. Many methodological techniques are available to damage the hair cells for subsequent scientific study. A recent method utilizes topical application of an ototoxic drug to the round window membrane. The current study examines the pharmacokinetics of gentamicin in the inner ear of chickens following topical application to the round window membrane or a single systemic high dose given intraperitoneally. Chickens were given gentamicin topically or systemically and survived for 1, 4, 12, 24, or 120 h (controls at 4 and 120 h). Serum and perilymph samples were obtained prior to sacrifice and measured for gentamicin levels. Results revealed higher levels of gentamicin in the perilymph of topically treated chickens than systemically treated chickens, with significant amounts of gentamicin still present in both at the latest survival time of 5 days. As expected, systemically treated chickens had much higher levels of gentamicin in the serum than topically treated chickens. Advantages and disadvantages to each method of drug administration are discussed. PMID- 15357418 TI - Onset neurones in the anteroventral cochlear nucleus project to the dorsal cochlear nucleus. AB - Considerable circumstantial evidence suggests that cells in the ventral cochlear nucleus, that respond predominantly to the onset of pure tone bursts, have a stellate morphology and project, among other places, to the dorsal cochlear nucleus. The characteristics of such cells make them leading candidates for providing the so-called "wideband inhibitory input" which is an essential part of the processing machinery of the dorsal cochlear nucleus. Here we use juxtacellular labeling with biocytin to demonstrate directly that large stellate cells, with onset responses, terminate profusely in the dorsal cochlear nucleus. They also provide widespread local innervation of the anteroventral cochlear nucleus and a small innervation of the posteroventral cochlear nucleus. In addition, some onset cells project to the contralateral dorsal cochlear nucleus. PMID- 15357419 TI - Spatial tuning curves along the chick basilar papilla in normal and sound-exposed ears. AB - Intense sound exposure destroys chick short hair cells and damages the tectorial membrane. Within a few days postexposure, signs of repair appear resulting in nearly complete structural recovery of the inner ear. Tectorial membrane repair, however, is incomplete, leaving a permanent defect on the sensory surface. The consequences of this defect on cochlear function, and particularly frequency analysis, are unclear. The present study organizes the sound-induced discharge activity of cochlear nerve units to describe the distribution of neural activity along the tonotopic axis of the basilar papilla. The distribution of this activity is compared in 12-day postexposed and age-matched control groups. Spontaneous activity, tuning curves, and rate-intensity functions were measured in each unit. Discharge activity at 60 frequency and intensity combinations was identified in the tuning curves of hundreds of units. Activity at each of these criterion frequency/intensity combinations was plotted against the unit's characteristic frequency to construct spatial tuning curves (STCs). The STCs depict tone-driven cochlear nerve activity along the length of the papilla. Tuning sharpness, low- and high- frequency slopes, and the maximum response were quantified for each STC. The sharpness of tuning increased with increasing criterion frequency. However, within a frequency, increasing sound intensity yielded more broadly tuned STCs. Also, the high-frequency slope was consistently steeper than the low-frequency slope. The STCs of exposed ears exhibited slightly less frequency selectivity than control ears across all frequencies and larger maximum responses for STCs with criterion frequencies spanning the tectorial membrane defect. When rate-intensity types were segregated, differences were observed in the STCs between saturating and sloping-up units. We propose that STC shape may be determined by global mechanical events, as well as localized tuning and nonlinear processes associated with individual hair cells. The results indicated that 12 days after intense sound exposure, global and local contributions to spatially distributed neural activity are restored. PMID- 15357421 TI - Acute endolymphatic hydrops generated by exposure of the ear to nontraumatic low frequency tones. AB - Low-frequency sounds presented at high nontraumatizing levels induce temporary hyperacusis in humans and animals. One explanation of this finding is that the basilar membrane operating point may be disturbed by an endolymph volume change. This possibility was investigated using volume and flow markers iontophoresed into the endolymphatic space of guinea pigs. Marker concentrations were measured with ion-selective microelectrodes placed apically and basally to the iontophoresis site during exposure of the ear to low-frequency tones. Concentration changes were interpreted quantitatively using a finite-element model of the endolymphatic space that allowed changes of endolymph cross sectional area and flow to be derived. Stimulation with a 200 Hz tone at 115 dB SPL for 3 min produced marker concentration changes consistent with the induction of transient endolymphatic hydrops and a basally directed displacement of endolymph. Endocochlear potentials were greater than normal after the exposure when hydrops was present. During identical tone exposures of animals without marker, we found that action potential (AP) threshold changes and endolymph potassium changes associated with the hydropic state were small. Marker concentration changes were compared with changes in endocochlear potential and AP thresholds for a range of exposure frequencies and levels. AP hypersensitivity occurred with 200 Hz exposure levels below those inducing endolymph volume disturbances. Endolymph volume changes are thought to be the result of, rather than the cause of, changes in operating point of the cochlear transducer. The observations that auditory threshold and endolymph potassium changes are minimal under conditions where substantial endolymphatic hydrops is present is relevant to our understanding of the hearing loss in patients with Meniere's disease. PMID- 15357423 TI - [Histologic pattern and mechanical properties of tissue-engineered tendon implants for tendon defects]. AB - This is a study on the histologic pattern and mechanical properties of tissue engineered tendon implanted for treatment of tendon defects. Tendons were resected from Roman chickens. Tendon cells were isolated from the tendons and cultured in vitro. The 2nd-4th passages of tendon cells were seeded on the degradable polyglycolic acid mesh to form cell-scaffold composites, which were further cultured for 7-10 days to construct tissue-engineered tendons. The tendon defects, 0.5 cm-0.8 cm in length, were made in the second digit flexor tendon bilaterally in 20 Roman chickens and then bridged with the constructed tissue engineered tendons. At 2 weeks, 4 weeks, 6 weeks, and 8 weeks post-operation, the samples of regenerated tendons were collected for gross examination, histologic staining and biomechanical test. After implantation of the tissue-engineered tendons, the wounds healed well. The gross appearance, the cells and collagen fibers arrangement of the regenerated tendons were similar to those of natural tendons, but there were relatively not many closely packed collagen fiber bundles organized in parallel with the tendons ("remodel"), so the maximum tensile force increased slowly and its value was 15.40+/-10.63 N at 8 weeks after surgery, reaching only 23% of that of natural tendon. The maximum strain was 22.49%+/ 10.21% at 8 weeks, being 10% higher than that of natural tendons. Polyglycolic acid scaffolds are degraded in vivo so rapidly that the regenerated tendons lose the normal biomechanical stimulus and then are unable to be remodeled. As a result, the mechanical strength of regenerated tendons is much lower than that of natural tendons. These results suggest that the normal biomechanical stimulus may be an important factor for the regenerated tendons to remodel. PMID- 15357420 TI - Isolation from cochlea of a novel human intronless gene with predominant fetal expression. AB - We have cloned a novel human gene, designated PFET1 (predominantly fetal expressed T1 domain) (HUGO-approved symbol KCTD12 or C13orf2), by subtractive hybridization and differential screening of human fetal cochlear cDNA clones. Also, we have identified the mouse homolog, designated Pfet1. PFET1/Pfet1 encode a single transcript of approximately 6 kb in human, and three transcripts of approximately 4, 4.5, and 6 kb in mouse with a 70% GC-rich open reading frame (ORF) consisting of 978 bp in human and 984 bp in mouse. Both genes have unusually long 3' untranslated (3' UTR) regions (4996 bp in human PFET1, 3700 bp in mouse Pfet1) containing 12 and 5 putative polyadenylation consensus sequences, respectively. Pfetin, the protein encoded by PFET1/Pfet1, is predicted to have 325 amino acids in human and 327 amino acids in mouse and to contain a voltage gated potassium (K+) channel tetramerization (T1) domain. Otherwise, to date these genes have no significant homology to any known gene. PFET1 maps to the long arm of human chromosome 13, in band q21 as shown by FISH analysis and STS mapping. Pfet1 maps to mouse chromosome 14 near the markers D14Mit8, D14Mit93, and D14Mit145.1. The human 6 kb transcript is present in a variety of fetal organs, with highest expression levels in the cochlea and brain and, in stark contrast, is detected only at extremely low levels in adult organs, such as brain and lung. Immunohistochemistry with a polyclonal antibody raised against a synthetic peptide to PFET1 sequence (pfetin) reveals immunostaining in a variety of cell types in human, monkey, mouse, and guinea pig cochleas and the vestibular system, including type I vestibular hair cells. PMID- 15357424 TI - [Simulation of inverse recovery of epicardial potentials under incomplete boundary conditions]. AB - Based on a 3-D inhomogeneous simulative body torso model, the influence of various boundary conditions on inverse recovered epicardial potential maps (EPM) was studied by boundary element method (BEM). The result shows that the precision of EPM calculated under incomplete boundary conditions will meet the clinical requirements, as long as the incomplete boundary conditions still contain the extremum area which often appears at the breast area. PMID- 15357422 TI - Null mutation of alpha1D Ca2+ channel gene results in deafness but no vestibular defect in mice. AB - Multiple Ca2+ channels confer diverse functions to hair cells of the auditory and vestibular organs in the mammalian inner ear. We used gene-targeting technology to generate alpha1D Ca2+ channel-deficient mice to determine the physiological role of these Ca2+ channels in hearing and balance. Analyses of auditory-evoked brainstem recordings confirmed that alpha1D-/- mice were deaf and revealed that heterozygous (alpha1D+/-) mice have increased hearing thresholds. However, hearing deficits in alpha1D+/- mice were manifested mainly by the increase in threshold of low-frequency sounds. In contrast to impaired hearing, alpha1D-/- mice have balance performances equivalent to their wild-type littermates. Light and electron microscope analyses of the inner ear revealed outer hair cell loss at the apical cochlea, but no apparent abnormality at the basal cochlea and the vestibule. We determined the mechanisms underlying the auditory function defects and the normal vestibular functions by examining the Ba2+ currents in cochlear inner and outer hair cells versus utricular hair cells in alpha1D+/- mice. Whereas the whole-cell Ba2+ currents in inner hair cells consist mainly of the nimodipine-sensitive current (approximately 85%), the utricular hair cells express only approximately 50% of this channel subtype. Thus, differential expression of alpha1D channels in the cochlear and utricular hair cells confers the phenotype of the alpha1D null mutant mice. Because vestibular and cochlear hair cells share common features and null deletion of several genes have yielded both deafness and imbalance in mice, alpha1D null mutant mice may serve as a model to disentangle vestibular from auditory-specific functions. PMID- 15357425 TI - [The effect of a simulated inflammation procedure in simulated body fluid on bone like apatite formation on porous HA/beta-TCP bioceramics]. AB - The formation of bone-like apatite on porous HA/beta-TCP bioceramics in dynamic simulated body fluid (SBF) undergoing a simulated inflammation procedure (pH = 6.5) was investigated in order to study the mechanism of osteoinduction and build a new method to choose biomaterials with better bioactivity. The results showed that the surface of porous HA/beta-TCP bioceramics which underwent a simulated inflammation procedure in dynamic SBF was more smooth. The light acidity in the simulated inflammation procedure would dissolve the fine grains and the parts possessing smaller curvature radius on the surface of porous HA/beta-TCP bioceramics, which would reduce the bioceramics solubility. Followed in normal SBF (pH = 7.4), the amount of bone-like apatite formed on the porous HA/beta-TCP bioceramics was less than that of porous HA/beta-TCP bioceramics incubation in normal SBF all along. The results also showed that the amount of bone-like apatite formed on the porous HA/beta-TCP bioceramics sintered by a microwave plasma was more than that of porous HA/beta-TCP bioceramics sintered by a conventional furnace. PMID- 15357426 TI - [Research of plasma adsorption and action of platelet adhesion of Dacron modified by plasma surface modification]. AB - In this paper, polyethylene glycol (PEG) of different molecular weight was grafted on the polyethylene terephthalate (PET, Dacron) films by plasma surface grafting modification. The competitive adsorption relation of plasma (fibrinogen and albumin) adsorbing on materials surface was analyzed in light of surface energy and interface free energy. The results indicated that the PET films grafted PEG long chain molecular possesses the characteristic of preferentially adsorbing albumin and this adsorption tendency of grafted PEG6000 sample is most distinct. The platelet adhesion tests of the PET films whose surfaces were pre set in contact with fibrinogen and albumin indicated that the surface adsorbing albumin can distinctly inhibit platelet adhesion and aggregation and possess favorable blood compatibility, but the surface adsorbing fibrinogen can enhance platelet adhesion and aggregation. PMID- 15357427 TI - [Effects of encapsulated plasmid recombining with hANP cDNA transfected cells on morphological and histological characteristic of experimental hypertensive rats]. AB - A technique based on release of the human atrial natriuretic peptide (hANP) from plasmid hANP cDNA transfected Chinese hamster ovary (CHO) cells encapsulated in polycaprolactone (PCL)-capsules was used for a potential therapeutic approach to hypertension or congestive heart failure (CHF). The plasmid combining with hANP cDNA was transfected into CHO cells, and then encapsulated plasmid hANP cDNA transfected CHO cells were implanted into two-kidney, one-clip (2K1C) hypertensive rats intraperitoneally. The morphological changes, histological changes were investigated after the implantation of PCL-capsules in 2K1C hypertensive rats. The results showed that the implantation of encapsulated hANP producing cells caused a significant delay of blood pressure (BP) increase after the encapsulated cells being implanted in 2K1C hypertensive rats. These effects were reflected morphologically by an attenuation of the glomerular sclerotic lesions, tubular damage and renal arterial thickening, comparing with control group. The plasma levels of hANP in 2K1C rats implanted with the PCL-capsules containing hANP-producing cells were higher than that of the control rats. These results demonstrated the usefulness of encapsulated hANP gene transfected cells as a new tool for hANP gene delivery in studying renovascular hypertension and cardiovascular diseases, thus implying the potential of using gene transfected cells as therapeutic agents in the treatment of cardiovascular diseases. PMID- 15357428 TI - [Effects of steep pulsed electric fields on cancer cell proliferation and cell cycle]. AB - To assess study the cytocidal and inhibitory effects of steep pulsed electric fields (SPEFs) on ovarian cancer cell line SKOV3, the cancer cell suspension was treated by SPEFs with different parameters (frequency, pulse duration, peak value of voltage). Viability rate and growth curves of two test groups (high dosage and low dosage of SPEFs) and one control group were also measured. The DNA contents and cell cycle were analyzed by flow cytometry (FCM). Different dosing levels of SPEFs exerted obviously different effects on cancer cell viability. With the enhancement of each pulse parameter, the viability rate was promoted and the inhibitory effect on the proliferation of treated cells was more evident. The cells exposed to SPEFs grew slower than the control. The ratio of S+G2/M phase cells was decreased, which restrained the DNA synthesis and division, but the ratio of G0/G1 phase cells was increased in the treated groups. It was also indicated that the SPEFs blocked the cell transition from G0/G1 phase to S+G2/M phase. There was a significant difference in cell cycle between treated group and control group (P<0.01). Lethal effects of SPEFs were represented by inhibiting the cancer cell proliferation at the cell level and by influencing the cell cycle at the DNA level. PMID- 15357429 TI - [Analysis of correlation degree between phase components and adsorptive amounts of serum proteins for DLC film]. AB - In the present paper, the influence of carbon phase components of three kinds of diamond like carbon (DLC) films, viz. DLC, DLC rich in graphite and DLC rich in diamond films, on adsorption of human serum albumin (HSA), human serum fibrinogen (HFG) and immunoglobin (IgG) was quantitatively analyzed by use of T-type correlation degree in the grey system theory. Through the analysis, the rational explanation for adsorptive amounts variations of the serum proteins with phase components in the experiment is reached and some essential conclusions have been obtained: (1) The effect of graphite phase and C-H phase on HSA adsorption are greater than that of other phase components; with the increase of these two phase coumponents, the adsorptive amounts of HSA decrease; (2) The powerful influence on HFG adsorption stems from DLC phase and C-O phase; with the decrease of DLC phase or the increase of C-O phase, the adsorptive amounts of HFG increase; (3) All of the carbon phase components have only limited influence on IgG adsorption in positive or negative fashion with a little difference in degree; (4) DLC phase has both effects of enhancing adsorption for HSA and weakening adsorption for HFG and IgG, thus its influence on the hemocompatibility of DLC films is much more important than that of other phase components. PMID- 15357430 TI - [Investigating the complexity of heart rate variability during anesthesia]. AB - By use of the complexity analysis indices (approximate entropy, complexity), the heart rate variability signals obtained from 38 subjects' ECG during anesthesia were analyzed. The results showed that there is an obvious chaos change of heart rate variability during anesthesia, both the complexity and approximate entropy of heart rate variability during anesthesia are evidently less than those during consciousness (P<0.05). In this dissertation, we decompose the heart rate variability during anesthesia into 1/f part and non-fractal part, and then analyze these parts. The results reveal that the sensitivity of the complexity indices can be improved by decomposing the heart rate variability. PMID- 15357431 TI - [The numerical simulation of pulsatile flow in a tapered blood vessel]. AB - The tapered blood vessel is associated with the human physiological pulsatile blood flow in this study to address the questions about the developing flow. The geometry model of tapered blood vessel, theoretical models of blood flow, physiological boundary conditions and calculation conditions are proposed. An average blood flow function is established according to the physiological pulsatile flow conditions. The pulsatile flow in a 3-D tapered blood vessel is simulated numerically with the flow function. The distributions of velocity and pressure on different time of the cardiac cycle are obtained. The numerical simulation outcomes are compared with the experimental and analytical results. The characteristics of the pulsatile flow in a tapered blood vessel are discussed in this paper. PMID- 15357432 TI - [Influence of wall thickness on the stress distribution within transtibial monolimb]. AB - Monolimb is a new type of lower-limb prostheses made of macromolecule polymer, in which the socket and prosthetic shank are integrative. Compared with traditional prosthesis, monolimb is more economical, good-looking and portable, so it indicates a possible direction in the future. Biomechanical research on trans tibial monolimb is necessary and helpful just like traditional prosthesis. In this article, a 3D FE model based on real geometry shape of an endoskeletal trans tibial monolimb is established. Keeping the same geometrical shape, three 3D FE models of transtibial monolimbs with different wall thickness are established. The influence of wall thickness on the stress distribution is analyzed under the load corresponding to the subphase of stance of Heel Off. The results indicate that stress within transtibial monolimb and pressure on the surface of soft tissue could be decreased with wall thickness of transtibial monolimb increased. This study will be helpful for the standard of wall thickness in designing transtibial monolimb. PMID- 15357433 TI - [Insight into surface structure and hemocompatibility of fluorinated poly(ether urethane)s and poly(ether urethane)s blends]. AB - It has been well known that fluorinated polyurethanes exhibit unique low surface energy, biocompatibility, biostability and nonsticking behavior. Consequently, these polymers have attracted considerable interest. In this study, the effect of various concentrations of fluorinated polyurethanes in the polyurethanes on the surface structures of the blends and their hemocompatibility were investigated by XPS, AFM, contact angle and platelet adhesion. It was found that the high concentration fluorine on the outer surfaces of the blends obtained with the low concentration of fluorinated polyurethanes (F: 0.342 wt%) in the blends was the same as that of the fluorinated poly(ether urethane)s, and all of the blends and the fluorinated poly(ether urethane)s had good hemocompatibility, compared with poly(ether urethane)s. The polymer blends and fluorinated poly(ether urethane)s suppressed platelet adhesion due to their high hydrophobicity and low surface tension. The XPS, AMF and contact angle results indicated that the high hydrophobicity of outer surface of the polyurethane blends is independent of the fluorinated polyurethanes content in the polymer blends but related to the concentration of the CF3 groups because the lower critical surface tensions and higher contact angle of many fluorinated surfaces reflect the concentration of CF3 groups. PMID- 15357434 TI - [Preparation of valaciclovir loaded bovine serum albumin nanoparticles surface modified with glycyrrhizin and its characteristics of targeting to liver]. AB - The valaciclovir was used as the model drug, the bovine serum albumin nanoparticles (BSA-NP) were prepared by desolvation process. Glycyrrhizin (GL) was oxidized by sodium periodate to be conjugated to surface reactive amino groups (SRAG) of the VACV-BSA-NP. Gel filtration method combined with HPLC method verified that GL was covalent coupling to the surface of VACV-BSA-NP with mean 9 GL residues per albumin molecule. The mean diameter of the VACV-BSA-NP-GL was 268 +/- 23 nm, the drug loading was 1.35%, and embedding ratio was 68.76%. The characteristics of release in vitro were in accord with two-phase kinetics. The uptake amount of VACV-BSA-NP-GL by primary cultured rat hepatocytes in vitro was higher, compared to the control-VACV-BSA-NP. 69.89% and 64.82% of the VACV were concentrated in liver at 15 min after i.v. VACV-BSA-NP-GL and VACV-BSA-NP, respectively. There is a significant difference between surface-modified group and control group (P<0.10). VACV-BSA-NP-GL was successfully prepared, which is considered to be a novel drug delivery system for targeting to hepatocytes. PMID- 15357435 TI - [The effect of fluid shear stress on the NO synthesis of rat osteoblast-like cells]. AB - This study sought to elucidate the function of NO during the signal transduction wherein fluid shear stress regulates the proliferation and differentiation of osteoblast cells. The isolated rat osteoblast-like cells were exposed to fluid shear stress 12 dyn/cm2 for 5, 10, 15, 30, 60 and 120 min respectively with the use of a flow chamber. The NO release was examined. After the exposure to fluid shear stress, the NO synthesis of rat primary osteoblast-like cells increased significantly (P<0.05) when compared with the control. After 60 minutes of exposure, the release of NO began to increase significantly (P<0.05), but no significant increase as such was seen in the control (P>0.05). NO synthesis may be one of the signal transduction pathways which transduce the fluid shear stress into osteoblast cells. In early stage, it may be induced by cNOS and in late stage by iNOS. PMID- 15357436 TI - [Construction of guided bone regeneration membrane by tissue engineering in vitro]. AB - In this study, porous polymer (PLA/PCL) membrane was first treated with ethanol to become hydrophilic, and then immersed into DMEM with 50% fetal bovine serum to enhance the affinity to cells. MSCs cultured in osteogenic medium were loaded into the membrane at density of 5 x 10(6)/cm2 for 7 days, and scanning electrical microscope was used to observe the growth of the MSCs. The growth of MSCs inside the constructs was functionally well, and the cells proliferated with the time of culture. We concluded from current study that the membrane had satisfactory biocompatibility and the constructs could be used to guided bone regeneration. PMID- 15357437 TI - [Synthesis and application of the polyacrylamide beads acting as LDL adsorbent's matrices]. AB - This study in pursuit of the synthetic technologies and structure characterization of polyacrylamide-based matrices (PAM beads) for low density lipoprotein (LDL) adsorbent and their adsorbability for LDL was intended for an experimental evidence of developing advanced matrices for LDL adsorbent. PAM beads were synthesized by inverse suspension polymerization, and their structure characterization was characterized by SEM, image analyzer and small angle X-ray scattering. The tripeptide serine-aspartic-glutamic acid (SDE) was coupled on the PAM beads to prepare the LDL adsorbents whose adsorbability for LDL was determined in vitro. The results showed that the PAM beads with the average size diameter 142.1 microm and the average pore diameter 119.8 nm could act as the matrices in accordance with the requirement of adsorbent for LDL. When the amount of acrylamide and the crosslinking agent N,N'-methylene-bis(acrylamide) was fixed, the average pore diameter decreased with the increase of the crosslinking agent content. Although the nonspecific binding of PAM beads for LDL was low, they could selectively adsorb LDL after coupling the SDE on the PAM beads. PMID- 15357438 TI - [The preparation and properties of Modified silk fibroin membranes by chitosan]. AB - The Modified silk fibroin membranes were prepared by mixing the aqueous solutions of both silk fibroin and chitosan with the use of oxidized glucose aldehyde as a crosslinking agent. It was characterized by FTIR, DSC, measurements of membrane potential and mechanical properties, the water swelling ratios and permeability coefficient for model drug 5-Fu in the different pH buffer solutions. It was shown that there were some strong hydrogen bond interaction and good compatibility between silk fibroin and chitosan molecules in the modified silk fibroin films. The isoelectric point of modified fibroin film was about pH 5.35, but that of natural fibroin film was around pH 4.5. It was also found that the mechanical properties of modified fibroin films were much better than those of fibroin itself. Its tensile strength and breaking elongation were greatly enhanced with the increase of chitosan content and their maximum values were as high as 71.4-72.7 MPa and 2.96%-3.82% respectively, at the composition of 40 wt% 60 wt% chitosan. Its coefficient of permeability decreased firstly and then increased slowly with the change of the pH value of solutions from pH 5 to pH 9, and the minimum coefficient of permeability was observed when pH=7. PMID- 15357439 TI - [Purification, induced differentiation and identification of rat embryonic neural stem cells]. AB - The methods of purification, expanding, marking, conservation, induced differentiation and identification of neural stem cells (NSCs) in vitro were explored for further research and treatment of tethered cord syndrome in children and other neural system diseases. The cells derived from the cerebral cortex of frontal lobe in 14.5 d rat embryos were maintained in serum-free DMEM/F12 medium containing 20 ng/ml bFGF, 20 ng/ml EGF and B27 supplement. The NSCs of suspending single-cell-colon were isolated and passaged, were purified by limited dilution, and were expanded numerously with sub-colon in consecutive generations. Then, Nestin antigen expression was detected by immunohistochemistry techniques. The cells of the purified and expanded NSCs were frozen, recovered and incubated in BrdU, and the NSCs were induced to differentiate in serum or feeder layer. These revived NSCs from frozen cells could express Nestin antigen and could be induced in serum or feeder layer to differentiate into neurons and glias expressing tubulin-III and GFAP respectively. It is good and simple for purification and proliferation of NSCs numerously by the limited dilution and consecutive generations suspending single-cell-colon of NSCs from the cerebral cortex of frontal lobe in rat embryos. The NSCs could be induced on feeder layer to differentiate into neural cells numerously. BrdU can mark and trace NSCs for the research and treatment of the animal model of neural system diseases. By good command of the technlogies for the purification proliferation, and induced differentiation of NSCs in vitro, it is possible to find a new way for further research of the biologic specificity and the treatment of the disease in nervous system. PMID- 15357440 TI - [Study on the modification surface material of heparinized polyurethane]. AB - In this study cationic and heparinized polyurethanes (PUs) were synthesized by a two-step solution polymerized method. Cationic and heparinized PUs were investigated by infrared spectroscopy, electron spectroscopy for chemical analysis (ESCA) and turbidity method. At the same time, the PUs proved of good biocompatibility through the laboratory tests, including blood coagulation time (CT), activated partial thromb plastin time (APTT) and fibroblast culture. These materials have good biocompatible function. PMID- 15357441 TI - [Adrenomedullin enhances glucose toxicity to pancreatic beta-cell of islets from spontaneously hypertensive rat (SHR)]. AB - This animal experiment was aimed at the questions whether high glucose concentration inhibits insulin secretion (glucose toxicity, GT) of beta-cell of islets from SHR and Wistar-Kyoto (WKY) rat and whether adrenomedullin (AM) enhances GT. Ten 6-week-old SHRs (test group) and ten 10-week-old Wistar-Kyoto rats (WKY) (control group) were selected. RAMI-1640 medium containing 5.6 mM glucose (normal glucose group) and 20 mM glucose (high glucose group) were applied. Various concentrations of AM (0, 10(-8), 10(-7), 10(-6) M) and RPMI-1640 medium containing high glucose were mixed, respectively. The isolated islets from rats were put into 12-well plates (90 islets/well). The islets were incubated in RAMI-1640 medium containing normal or high glucose for one hour. Then the supernatants from both incubations were determined by RIA for insulin. In SHR group, the insulin concentration in supernatants gained from high glucose group without AM was lower than that from normal glucose group (19.9+/-6.6 vs 60.9+/ 33.6 mU/L, P<0.05). With the increment of the concentration of AM, insulin concentration in supernatants from islets incubated in high glucose and various concentrations of AM tended to be low further (19.9+/-6.6 vs 22.2+/-8.0 vs 21.5+/ 5.6 vs 17.9+/-3.6 mU/L). The changing tendency in control group was the same as in SHR group. When the islets were incubated in normal glucose and high glucose medium, the insulin concentration in supernatant significantly decreased in SHR group compared with that in control group (P<0.01). The insulin secretion was inhibited by high glucose in beta-cell of islets from SHR and WKY. The results suggest GT to beta-cell of islets from SHR and WKY. AM tended to inhibit insulin secretion in a dose-dependent manner in beta-cell of islets from SHR and WKY. The inhibition of insulin secretion caused by high glucose in beta-cell of islets from SHR was more remarkable than from WKY. This may be related to secretion dysfunction in beta-cell of islets from SHR. PMID- 15357442 TI - [Transplantation of adrenocortical cells in allorat using porous PHB as cell carrier]. AB - This experiment was designed to investigate the feasibility of transplanatation of using porous PHB as cell carrier for the transplanatation of adrenocortical cells. Adrenocortical cells from rat adrenal gland were separated and cultured in vitro. The effect of PHB on the proliferation and secretory function of adrenocortical cells were evaluated by MTT and RIA methods. Then adrenocortical cells were seeded into porous PHB. After the cells were cultured in vitro for about seven days, they were implanted into the rats having undergone bilateral adrenalectomy. The changes of blood corticosterone and aldosterone and the local histological changes in these rats were observed. Adrenocortical cells were able to grow and survive on PHB. No effect on the proliferation and secretory function of adrenocortical cells were observed. Most bilateral adrenalectomized rats bearing the transplanted adrenocortical cells within PHB (study group) survived longer than did the adrenalectomized rats in control group. The blood corticosterone level and aldosterone level of study group were higher than those of control group. It was found that PHB has no effects on the survival, proliferation and secretory function of adrenocortical cells. Adrenocortical cells within PHB can survive a period of time and can secrete corticosterone and aldosterone which can meet the needs of the adrenalectomized rats. PHB can degrade slowly in vivo. It is feasible to perform transplantation of adrenocortical cells using porous PHB as cell carrier. PMID- 15357443 TI - [Preparation of heart valve scaffold and cell seeding]. AB - To prepare scaffolds for heart valve tissue engineering, porcine heart valves were treated with varied concentrations of trypsin for 32, 56, 80 and 104 h or followed with DNase. And then the structure of acellular valves was observed under light microscope, scanning and transmission electron microscope. Porcine endothelial cells, human endothelial cells, and canine myofibroblasts were reseeded onto the acellularized porcine heart valve scaffolds once a day for 3 days. The valves were analyzed by immunohistochemical staining and electron microscopy. Results show that all endothelial cells and the majority of interstitial cells were removed from the heart valves after digestion with trypsin for 104 h, and the collagen fiber structure remains intact, but the space between collagen fibers increased slightly. Incubation with trypsin for 80 h and then with DNase almost removed all cells, and the collagen fiber structure and the space between the fibers remain intact. After reseeding, human endothelial cells almost fully cover the valve scaffold surface as shown by H-E staining and platelet endothelial cell adhesion molecules (PECAM-1) staining. Xenogeneic porcine endothelial cells also adhered to and grew on the scaffolds. As shown by H-E staining and actin staining, canine myofibroblasts not only adhered to the surface of valve scaffold but also migrated to the inner part of matrix after one week culture. These results suggest that the digestion of porcine heart valves with trypsin combining with DNase is a suitable method to remove cells. The acellular porcine heart valve scaffolds have a quite favorable biocompatibility with human and porcine endothelial cells as well as canine myofibroblasts. PMID- 15357444 TI - [Necrosed field model of high intensity focused ultrasound for tumor treatment]. AB - High intensity focused ultrasound (HIFU) has become a new developed micro invasive surgical treatment of tumor, which is a hot topic in basic and applied research field. HIFU has shown its unique advantages including micro-invasion and curative effect during the treatment of several kinds of superficial tumor, e. g. mammary cancer. Based on the cellular thermo-necrotic theory, the expression of omega value was introduced to establish the mathematic model of the necrosed field formed in HIFU treatment progress. A non-interferential and self-focused system with multi-transducer was used to verify the theoretical model. Results of this study proved that the necrosed field mathematic model can simulate the actual heating progress. The model of this study may predict the practical value of the necrosed field as well as the time needed to take shape. PMID- 15357445 TI - [Simulation of skeletal muscles dielectric behaviour with theoretical model]. AB - The dielectric behavior of the frog skeletal muscle cells in the 100 Hz-100 MHz range was simulated with dielectric ellipsoidal-shell model, and the model's parameters were put forward for the first time. It could serve as a theoretical foundation on which to make analysis of muscle fatigue, muscle malnutrition and muscle atrophies in the future. PMID- 15357446 TI - [A biomechanical study on the morphological changing process of human bitemarks]. AB - This is a biomechanical research in the morphological changing process of human bitemarks on the skin of dogs. The human bitemarks were made on the live and dead dogs by different tooth and occlusal force. The changing process of the bitemarks were recorded for a long time and its related morphological parameters were measured. Then multiple stepwise regression analysis was made to disclose the relationship of the shape of the bitemarks to occlusal force, time, and tooth area, tooth width and thickness. The mathematic relationship of the morphological changing process of the bitemark with occlusal force, time and tooth shape was established. PMID- 15357447 TI - [Research on DPOAE of guinea pigs under awake and anesthetic conditions]. AB - In order to observe the DPOAE of guinea pigs under awake and anesthetic conditions, we invesitigated the amplitude of distortion product emission and I/O function curves and made a comparative analysis of their changes under different conditions. It was found that the amplitude of DPOAE and I/O curves did not show difference in the guinea pigs under awake and anesthetics conditions. We deem it credible and simple to measure DPOAE in the guinea pigs under awake condition, which is similar to measuring DPOAE in the guinea pigs under physiological condition. PMID- 15357448 TI - [Cell apoptosis in atrophic skeletal muscle induced by immoblization in rabbits- an experimental study using TUNEL]. AB - This experiment was designed to explore the correlation between the mechanism of immobilization-induced skeletal muscle atrophy and the apoptosis of muscular cells. The models of skeletal muscle atrophy induced by immobilization for different length of time were established according to Sievanen II methods. 24 rabbits, each of them having one hind leg fixed by the tubal plaster and the other one free as control, were randomly divided into four groups depending on time of fixation (3, 7, 14, and 28 days respectively). The animals were sacrificed by the end of fixation. TdT-mediated d-UTP nick end labeling (TUNEL) was used to investigate the apoptotic muscle cells in the animal's bone. By comparing the apoptotic muscle cells with the morphology of the skeletal muscle, the correlation between cell apoptosis and skeletal muscle atrophy were analyzed. Apoptotic muscle cells did appear after immobilization in the atrophied skeletal muscle. In various groups, some cells with false positive stained TUNEL were found in the atrophic muscle, which could be distinguished from apoptotic cells by their characteristics. In conclusion, cell apoptosis participates in the process of skeletal muscle atrophy induced by immobilization; the amount of apoptotic cells is strongly associated with the time of immobilization, its peak appears on the 14th day of immobilization; the distribution of apoptotic skeletal muscle cell varies with the time of fixation. The severity of skeletal muscle atrophy is associated with the degree of the muscle cell apoptosis. PMID- 15357449 TI - [Application of enzyme histochemistry in evaluation of in vitro and in vivo biocompatibility of HA/TCP]. AB - In order to explore the possibility of applying enzyme histochemistry in biocompatibility evaluation, we investigated the effect of biomaterials on the activities of intracellular enzymes in this experiment. It was found that there was no obvious difference in morphology between osteoblasts co-cultured with HA/TCP and with Ti-alloy. However, transient down-regulation of NADH, SDH, LDH and CCO of the osteoblasts co-cultured with HA/TCP was detected by enzyme histochemistry, but these enzymes of osteoblasts the co-cultured with Ti-alloy were not down-regulated. It was indicated that something extracted from HA/TCP injured the co-cultured osteoblasts slightly. Similar early acute inflammatory reactions were observed after HA/TCP and Ti-alloy were separately implanted into the dorsal muscle of rabbit. There was also no obvious difference between the tissue response to HA/TCP and that to Ti-alloy. Activities of enzymes in tissues around implanted materials were down-regulated at the early injury period and recovered gradually within 30 days post-operation. But the mild toxicity of extracts from HA/TCP was demonstrated by the fact that the recovery period of HA/TCP group was longer than that of Ti-alloy group. It was proved that enzyme histochemistry is more sensitive than tissue morphology analysis in detecting the cell or tissue responses to biomaterials. Therefore, it is possible to use enzyme histochemistry in biocompatibility evaluation. PMID- 15357450 TI - [Using AR model to analyze injured nerve with needle EMG signal]. AB - The two main factors to affect the style of the recruitment are temporal recruitment and spatial recruitment. This study sought a new way to analyze the recruitment with the modern spectrum method. The abnormal spatial recruitment and temporal recruitment of varied injury degrees of intramuscular neuron were compared through the AR model. At last, AR coefficients were extracted and passed through BP artificial neuron network to classify different NEMG signals and good result was gained. PMID- 15357451 TI - [A simulating experimental system for interventional treatment of lung cancer]. AB - In order to go deeply into the researches on the theory of hemodynamics for interventional treatment of lung cancer, we have designed a simulating experimental system. Using the experimental system, we can simulate the operation process and test the effect of drug infusion under a variety of physiological parameters. The parameters that admit of adjustment are: cardiac output, blood pressure of aorta, waveform of pressure, pulse period, blood flow in bronchial artery and different types of catheter, injecting velocity, injecting technique, etc. In addition, the entrance position and blood vessel diameter of small embranchment artery can be changed. In this paper are presented the application of our pH-testing method to natural infusion region and the determined representational shape and size. PMID- 15357452 TI - [Power spectral analysis on heart rate variability of hypoxaemia in fetal lambs]. AB - To analyze the effect of hypoxaemia on heart rate variability (HRV) in fetal lambs by means of power spectrum, the intrauterine surgical operations were performed at 116-125 gestational days in 7 lambs. Arterial catheter was inserted in the fetal femoral artery and sent to aorta abdominalis, and blood pressure was recorded continually on tape recorder. The microspheres were injected via the arterial catheter to block the micrangium of placenta, thus making an animal model of fetal hypoxaemia. The fetal blood sample was drawn through the catheter for blood gas analysis. In terms of the heart beat variability power spectral density, there were four consistent components, namely very low (VL, 0.01-0.025 cycle/beat), low (L, 0.025-0.125 cycle/beat), middle (M, 0.125-0.2 cycle/beat), and high (H, 0.2-0.5 cycle/beat). Integrated peaks in the power spectrum were compared before and after administration of microsphere. The spectral power in the L frequency components was significantly increased (0.07 +/- 0.01 vs. 0.21 +/ 0.03, P<0.01), and the spectral power in the H frequency components was significantly reduced (0.53 +/- 0.1 vs. 0.27 +/- 0.05, P<0.05). There was no significant difference in M and VL. The times of microsphere injection were related to fetal blood pH (r=0.585, p<0.01), PCO2 (r=0.5, p<0.05) and PO2 (r=0.75, P<0.01). The results clearly demonstrate the association between change of power spectrum of heart rate variability and the effect of hypoxia of the fetus in labour. PMID- 15357453 TI - [Development and research of temporary demand pacemaker with electrocardiosignal display]. AB - A temporary demand pacemaker with electrocardiosignal display is introduced in this paper. Double way low-noise electrocardiosignal preamplifier, amplitude limiter, high and low pass filter, 50 Hz notch filter, TTL level generator and stimulating pulse formation circuit are components of the hardware electrocircuit. The demand pacing and the electrocardiosignal display are separately controlled by the software in which the double microcontrollers communications technique is used. In this study, liquid crystal display is firstly used in body surface electrocardiosignal display or intracardial electrophysiologic signal display when the temporary demand pacemaker is installed and put into use. The machine has proven clinically useful and can be of wide appliation. PMID- 15357454 TI - [Method for measuring different phases of left ventricular systole period with ultrasonic Doppler and synchronism electrocardiogram in normal persons]. AB - This study of methodology was aimed to assess the feasibility of measuring different phase of left ventricular systole period with ultrasonic Doppler and synchronism electrocardiogram. We measured the phases of left ventricular systole period and left ventricular volumes in 13 normal persons, using spectrum of aortic valve orifice flow and synchronism electrocardiogram. The values measured by routine method and by Doppler method were compared. The correlation and agreement between the measures ascertained by the two ways were analyzed. The time parameters of end systole of left ventricule showed no significant difference between the two ways (P>0.05); there was high linear correlation between the two parameters (r=0.91, P<0.01); there were good agreements between the two parameters of the two ways by Bland-Altman analysis. The volume parameters of left ventricular systole end showed no significant difference between the two ways (P>0.05); there was high linear correlation between the two parameters (r=0.97, P<0.01); there were good agreements between the two volume parameters of the two ways. The volume in the left ventricular systole end determined by Doppler method and the volume in the left ventricular diastole end determined by the routine method showed no significant difference (P>0.05); there was high linear correlation between the two parameters (r=0.98, P<0.01); there was good agreement between the two volume parameters of the two ways. It is feasible to measure different phases of left ventricular systole period with the combined use of ultrasonic Doppler and synchronism electrocardiogram, and this combinaterial method and the routine method can be replaced with each other according to the clinical setting. PMID- 15357455 TI - [Numerical simulation of thyroxin producing process]. AB - Numerical simulation of the thyroxin producing process of a human body is presented to analyze the dynamic and static variation laws of the process. The relationship between the static value of thyroxin and the reaction coefficients of various physiologic chemical reactions upon the producing thyroxin is clarified. The simulation results provided in this paper can be utilized as some reference data for clinical pathogeny analysis of some thyroxin diseases. PMID- 15357456 TI - [Sonication of biomaterial and the regulation of acoustic parameters]. AB - The acoustic properties of ultrasound and the interaction of biomaterial and cavitation are analyzed. The relation between ultrasonic parameters and sonication is indicated. Our research revealed that different sonication aims must well match with different acoustic properties for optimizing the sonication technology. Based on the theory of wave superposition, a method for enhancement of ultrasonic intensity in wide dimension is introduced. A large scale powerful polyhedral acoustic field is built according to the research above. The rationality and effectiveness of the method are demonstrated through examination. PMID- 15357457 TI - [Custom design of hip joint prostheses based on X-ray films]. AB - A novel method for the design of hip joint prostheses based on X-ray films is introduced. Only arcs and straight-lines form figures of hip joint prostheses. Because geometrical tolerances of manufacturing hip joint prostheses matching section are usually not strict, hip joint prostheses can be manufactured without CNC machine tool. Three hip joint prostheses for three different femurs were designed through a program which was developed by the present authors. Approximate marrow cavities of these femurs were simulated according to a standard database about femur. These models of femur marrow cavities were used to verify the hip joint prostheses designed. These hip joint prostheses designed were manufactured and implanted into femurs respectively. Experimental results indicate that the novel method for the design of hip joint prostheses is practicable. PMID- 15357458 TI - [A computer-aided image diagnosis and study system]. AB - The revolution in information processing, particularly the digitizing of medicine, has changed the medical study, work and management. This paper reports a method to design a system for computer-aided image diagnosis and study. Combined with some good idea of graph-text system and picture archives communicate system (PACS), the system was realized and used for "prescription through computer", "managing images" and "reading images under computer and helping the diagnosis". Also typical examples were constructed in a database and used to teach the beginners. The system was developed by the visual developing tools based on object oriented programming (OOP) and was carried into operation on the Windows 9X platform. The system possesses friendly man-machine interface. PMID- 15357459 TI - [The research progress in ligament tissue engineering]. AB - Ligament injury always has an unsatisfied outcome because of the poor blood supply and scar tissue formation. This may result in severe joint dysfunction. Tissue engineering, as a most prospective field, may provide an effective approach for the treatment of ligament injury. This paper has reviewed some recently published articles focusing on the sources of seed cells in ligament tissue engineering, application of growth factors, screening of scaffold materials with specific mechanical and biodegradable properties, and interaction between cells and scaffold materials. At present, what should be extensively studied are scaffolds with specific mechanical and biodegradable properties, and bioreactors providing three-dimensional culture microenvironment mimic in vivo. PMID- 15357460 TI - [Knowledge discovery in database and its application in clinical diagnosis]. AB - Nowadays the tremendous amount of data has far exceeded our human ability for comprehension, and this has been particularly true for the medical database. However, traditional statistical techniques are no longer adequate for analyzing this vast collection of data. Knowledge discovery in database and data mining play an important role in analyzing data and uncovering important data patterns. This paper briefly presents the concepts of knowledge discovery in database and data mining, then describes the rough set theory, and gives some examples based on rough set. PMID- 15357461 TI - [Advance of fast magnetic resonance spectroscopic imaging]. AB - The required time of conventional magnetic resonance spectroscopic imaging technique is too long to be applied to clinic. It is necessary to develop the fast methods for magnetic resonance spectroscopic imaging. Nowadays there are 7 kinds of methods presented, which come from MRI techniques. In this contribution the conventional spectroscopic imaging and 7 sorts of fast spectroscopic imaging are elaborated. It is envisaged that more rapid imaging techniques will be designed, if these arbitrary trajectory reconstruction methods in MRI are applied to spectroscopic imaging. PMID- 15357462 TI - [Progress of researches on carbon/carbon composites used in human loaded bones]. AB - Carbon/carbon composites have excellent biocompatibility with human hard tissue and elasticity modulus similar with that of human bones, which endow them great potential applications in substitution for human loaded bones. The current research situations and applications of carbon/carbon composites in human loaded bones are reviewed. The coating technologies of bioactive layers on carbon/carbon composites are discussed. The problems to be solved and the prospects of carbon/carbon composites in human loaded bones are analyzed and predicted. It is believed that bioactive layers coating on carbon/carbon composites should play an important role in human loaded bones. PMID- 15357463 TI - [Visualization technology and its application in medical image]. AB - There are many noise disturbances in ultrasound image, and the surgical treatments are in great request for medical image, but the three dimensional image based on traditional surface graphics method cannot satisfy the request. Nevertheless, researches on visualization technology involves the denotation, operation and vision of the volume data the object contains, and the volume data contains ampler and more intact information than does the surface, therefore visualization technology will be widely used in the field of three dimensional medical image. PMID- 15357464 TI - [The progress of algorithms applied in digital hearing aid]. AB - The paper introduced the structure of digital hearing aid in brief firstly, then analyzed and compared signal processing algorithms applied in digital hearing aid, serving respectively in multi-channel frequency compensation, noise reduction and acoustic feedback cancellation. Finally, several special signal processing techniques used in digital hearing aid were introduced. PMID- 15357465 TI - Delaying Alzheimer's. In a new study, Aricept held it off for 18 months. Is this drug right for you? PMID- 15357466 TI - The price of pressure. Not all stresses are created equal. A new study finds that some may even be good for you. PMID- 15357467 TI - Back on the AIDS alert. Once a model in the war against HIV, Thailand dropped its guard. Now the country's at risk again. PMID- 15357468 TI - Allergic to wheat? Millions are without knowing it--but a simple test can help you find out. PMID- 15357469 TI - Secrets behind the mask. PMID- 15357470 TI - [Chaos--workoholism--writer's block. Occupational disorders in physicians]. PMID- 15357471 TI - [Participate in the large stroke action: identify the risks!]. PMID- 15357472 TI - [Diagnosis at a glance in metabolic diseases: when skin symptoms indicate diabetes]. PMID- 15357473 TI - [Bulimia and anorexia. Cannot be explained by psychosomatic factors alone]. PMID- 15357474 TI - [Do the best physicians work only for private patients? No, for bureaucracy!]. PMID- 15357475 TI - [No motivation for "hunger diets and exercise terror". How to motivate diabetic patients for a healthier life style? (interview by Dr. Maria Weiss)]. PMID- 15357476 TI - [Chronic abdominal pain--internistic-psychosomatic aspects]. AB - Abdominal pain is considered to be chronic when it persists for at least three months or when a patient experiences such pain for a total of three months during the course of a year. Pathophysiologically, nociceptive/neuropathic functional pain syndrome, mental disorders with the cardinal symptom of chronic pain, and mixed forms can be distinguished. In 50% of the patients, the cause of chronic abdominal pain is a functional gastrointestinal disorder e.g. functional dyspepsia irritable bowel syndrome. On the basis of a structured pain history, a physical examination and a basic "technical" diagnostic program (laboratory investigations, abdominal ultrasonography, Esophagogastroduodenoscopy, colonoscopy), correct assignment to one of the above-mentioned can be achieved in most of the cases. PMID- 15357477 TI - [Pelvic pain--definitions, diagnosis and therapeutic options]. AB - Depending on the location, bladder, urethral, penile and prostate pain syndromes are distinguished. In addition to the investigation of urine and ejaculate with the aim of detecting an infection, clinical examination, uroflowmetry are employed and, where indicated, imaging techniques to exclude cancer disease. Chronic pelvic pain is a non-cancer-related pain persisting for more than six months. In the absence of an underlying treatable disease, appropriate pain therapy can be initiated with antiphlogistic analgesics, COX 2 inhibitors or opioids. Depending on the clinical presentation, further therapeutic options or such non-drug measures as bladder training, dietetic measures or surgery may be applied in addition. PMID- 15357478 TI - [Gynecological aspects of pain in the lower abdomen]. AB - Lower abdominal pain in women may be a manifestation of disorders of various organ systems. In addition to surgical, urological, orthopedic, neurological, and psychogenic problems, gynecological causes have a major role to play. In the absence of other confirmed causes, women with acute or chronic lower-abdominal pain must therefore be submitted to a gynecological investigation. Important components of such an investigation are clinical gynecological examination and transvaginal ultrasonography. In women of childbearing age, a pregnancy test should always be performed. For both diagnostic and therapeutic purposes, laparoscopy is constantly gaining in importance. PMID- 15357479 TI - [Diagnosis of hypertension, an update--efficient and cost-saving]. PMID- 15357480 TI - [Diagnosis and treatment of acute diarrhea]. PMID- 15357481 TI - [Catamenial pneumothorax--a case report]. AB - Catamenial pneumothorax is a recurrent condition, predominantly affecting the right lung, of women of childbearing age, which is closely related to the menstruation cycle. Additional endometriosis and a defect of the right diaphragm is found relatively frequently. The case of a patient with a history of endometriosis, and a surgically confirmed defect of the diaphragm with prolapsed liver tissue, in whom pneumothorax first occurred at the age of 46, is presented. PMID- 15357482 TI - [Qualitatively reliable study proves: in chronic bronchitis or COPD--standardized Myrtol]. PMID- 15357483 TI - [General practice immunized against unpaid overtime. Colleague collects double value from vaccination]. PMID- 15357484 TI - [Federal judge on severe complications after vaccination. Patient handout counts as educational material]. PMID- 15357485 TI - [The new world of health care reform. Will you be a preferred provider or a standard provider of left-over work?]. PMID- 15357487 TI - [Normal blood pressure values in general practice are not enough. Also measure "morning after" values]. PMID- 15357488 TI - [Bedridden patients at high risk for decubitus ulcer. Skin oil prevents decubitus wounds]. PMID- 15357489 TI - [New combination therapy. Up to 60% lower LDL cholesterol level]. PMID- 15357490 TI - [Due to normal transaminase values, no further treatment. Hepatitis C patient "suddenly" has liver cirrhosis]. PMID- 15357491 TI - [After taking the table, immediate sex? New potency pill allows more time for love life]. PMID- 15357492 TI - [Chronic polyarthritis and neck pain]. PMID- 15357493 TI - [10 minute consultation: acute spontaneous vision loss]. PMID- 15357494 TI - [Minor manifestation, major etiology]. PMID- 15357495 TI - [Diagnostic and therapeutic challenge: deep venous thrombosis. Preventing pulmonary embolism]. PMID- 15357496 TI - [Every 5th person is allergic, more asthma fatalities than traffic deaths. How can the allergy epidemic be controlled?]. PMID- 15357497 TI - [Life threatening malnutrition is often overlooked. Are your elderly patients adequately nourished? (interview by Christine Vetter)]. PMID- 15357498 TI - [ACE inhibitor and beta blocker are indispensible in heart failure. With adverse effects consider concomitant medication first]. PMID- 15357499 TI - [Psychiatric disorders in children and adolescents]. PMID- 15357500 TI - [Anxiety disorders in children and adolescents]. AB - Anxiety disorders are among the most common psychiatric disorders in children and adolescents with estimated prevalence rates of between five and ten percent. Comorbidity rates are high--in particular for further anxiety and depressive disorders. The etiology is considered to be multifactorial, with genetic, biological and temperamental factors as well as stressful life events, parental role models and parenting all playing a part. Therapeutic programs combine psychological counseling--including the family--psychotherapy and psychopharmacotherapy, as required. As psychotherapy, cognitive-behavioral interventions have proven highly effective. In severe disorders that prevent the overcoming of complex anxiety/phobias (e.g. going to school impossible), psychopharmacotherapeutic measures can be helpful, and may support, or, in individual cases even make possible, successful psychotherapy. PMID- 15357501 TI - [Course and prognosis of obsessive-compulsive disorders]. PMID- 15357502 TI - [Long-term courses in children with attention deficit hyperactivity disorder (ADHD)]. AB - ADHD was long considered to be the attention deficit disorder of childhood. Observations over the long-term have, however, shown that some one-third of victims also continue to meet the criteria for ADHD in adulthood too. In many cases, such additional psychiatric conditions as depression or an addictive illness are also to be seen. Today, ADHD is believed to be a mainly heritable disorder with deviations in the dopaminergic system. Re-evaluation of the condition now focuses attention on the attention-related, rather than the hyperactivity-related aspects of the problem. Currently, the best therapeutic results are achieved through the use of stimulants coupled with intensive paediatric psychiatric care. PMID- 15357503 TI - [What's new in insulin treatment?]. PMID- 15357504 TI - [Elements of managed care in Germany and in Switzerland]. AB - With the introduction of the GVK (health insurance) modernization statute (GMG), health policy in Germany will focus on a more highly differentiated form of remuneration of the physician's services that is based on the degree of morbidity of the patient. Simultaneously with the introduction of morbidity-oriented risk structure adjustment in the year 2007, the level of payment to the doctor will be determined by the standard services volume based on risk categories and relative weightings as well as the case-based flat rates. A glance across the border to our Swiss neighbors provides information on their experience with diagnosis related forms of remuneration. PMID- 15357505 TI - [Thyroid scintigraphy]. PMID- 15357506 TI - [August Natterer exhibit in Heidelberg. Displays of the world court]. PMID- 15357507 TI - Temporal context in concurrent chains: I. Terminal-link duration. AB - Two experiments are reported in which the ratio of the average times spent in the terminal and initial links (Tt/Ti) in concurrent chains was varied. In Experiment 1, pigeons responded in a three-component procedure in which terminal-link variable-interval schedules were in constant ratio, but their average duration increased across components by a factor of two. The log initial-link response ratio was a negatively accelerated function of Tt/Ti. Overall, the data were well described by Grace's (1994) contextual choice model (CCM) with temporal context represented as (Tt/Ti)k or 2Tt/(Tt + Ti), and by Mazur's (2001) hyperbolic value added model (HVA), with each model accounting for approximately 93% of the variance. In Experiment 2, fixed-parameter predictions for each model were generated, based on the data from Experiment 1, for conditions in which Tt/Ti was varied over a more extreme range. Data were consistent with the predictions of CCM with temporal context represented as 2Tt/(Tt + Ti) and to a lesser extent as (Tt/Ti)k, but not with HVA. Overall, these results suggest that preference increases as a hyperbolic function of Tt/Ti when terminal-link duration is increased relative to initial-link duration, with the terminal-link schedule ratio held constant. PMID- 15357508 TI - Function transfer in human operant experiments: the role of stimulus pairings. AB - Although function transfer often has been studied in complex operant procedures (such as matching to sample), whether operant reinforcement actually produces function transfer in such settings has not been established. The present experiments, with high school students as subjects, suggest that stimulus pairings can promote function transfer in conditions that closely approximate those of matching to sample. In Experiment 1, the subjects showed transfer of operant responding from three geometric figures (C1, C2, C3) to three colored shapes (B1, B2, B3) when the latter were paired with the former. Experiment 2 involved two groups of subjects. In the matching group, subjects matched the colored shapes with the geometric figures; in the yoked group, the shapes were merely paired with the geometric figures, and the schedule of stimulus pairing was yoked to the performance of the subjects in the matching group. Both groups of subjects showed function transfer. Experiment 3 documented function transfer from C stimuli to B stimuli through indirect stimulus pairings (A-B, A-C). In Experiment 4, function transfer was obtained even though the subjects vocalized continuously during the pairing trials, presumably preventing covert verbalization that might mediate transfer effects. Our results are consistent with a Pavlovian account and raise difficulties for current operant theories of function transfer. PMID- 15357509 TI - The relation between stimulus function and equivalence class formation. AB - Fifty participants were exposed to a simple discrimination-training procedure during which six S+ functions were established for six arbitrary stimuli, and S- functions were established for a further six stimuli. Following this training, each participant was exposed to one of five conditions. In the S+ condition, participants were exposed to a stimulus equivalence training and testing procedure using only the six S+ stimuli as samples and comparisons. In the S+/S- condition, participants were exposed to the same training and testing sequence as in the S+ condition, the difference being that three S+ and three S- stimuli were used as sample and comparison stimuli, with each set of three corresponding to the trained equivalence relations. In the S+/S- mixed condition, the S+ and S- stimuli were assigned to their roles as samples and comparisons in a quasi-random order. In the S- condition, all six S- stimuli were used. The no-function condition served as a control condition and employed stimuli for which no stimulus-control functions had been established. The results showed that, on average, participants required more testing trials to form equivalence relations when the stimuli involved were functionally similar rather than functionally different. Moreover, participants required more test trials to form equivalence relations when novel arbitrary stimuli, rather than functionally distinct stimuli, were used as samples and comparisons. The speed of acquisition of stimulus equivalence was also related to the number of functionally similar stimuli established before training. These findings indicate a variety of ways in which the emergence of equivalence relations is affected by the functional classes in which the relevant stimuli participate. PMID- 15357510 TI - Naming and categorization in young children: II. Listener behavior training. AB - Following pretraining with everyday objects, 1- to 4-year-old children received listener training with three pairs of arbitrary stimuli of differing shapes. For each pair, 9 children were trained to select one stimulus in response to the spoken word /zog/ and the other to the spoken word /vek/. Next, in the look-at sample category match-to-sample test, none categorized the six stimuli correctly when asked to look at the sample before selecting from five comparisons. Seven of these children failed a subsequent test of corresponding speaker behavior (tact test); following tact training, 5 of them passed either a repeat of the look-at sample category test (2 subjects) or an alternative category test (3 subjects) in which they were required to tact the sample before selecting comparisons. The remaining 2 failed both category tests. Of the 2 who passed the tact test, 1 passed the tact-sample category test; the other failed to complete category testing. Two children were next given a second stimulus set. One passed the look at-sample category test and the tact test; the other failed both tests but passed the tact-sample category test after tact training. The results show that 1- to 4 year-old children may learn listener behavior without corresponding speaker behavior. The results also show that common listener behavior is not sufficient to establish arbitrary stimulus classes, and they are consistent with the proposition that naming may be necessary for categorization of such stimuli. PMID- 15357511 TI - Wiener filter estimation of transfer functions. AB - The use of a Wiener filter estimate for the linear transfer function can significantly improve the description of behavioral dynamics. This report presents a two-pass, Monte-Carlo-based algorithm that is well suited to repeated trials local average measurements. The Wiener filter transfer functions strongly suppress noise artifacts as well as allow reliable transfer function determination under a much wider class of reinforcement schedules. Implications of expanding the possible form of experimental design are considered along with improvements in the fidelity of resulting predictions. PMID- 15357512 TI - A computational model of selection by consequences. AB - Darwinian selection by consequences was instantiated in a computational model that consisted of a repertoire of behaviors undergoing selection, reproduction, and mutation over many generations. The model in effect created a digital organism that emitted behavior continuously. The behavior of this digital organism was studied in three series of computational experiments that arranged reinforcement according to random-interval (RI) schedules. The quantitative features of the model were varied over wide ranges in these experiments, and many of the qualitative features of the model also were varied. The digital organism consistently showed a hyperbolic relation between response and reinforcement rates, and this hyperbolic description of the data was consistently better than the description provided by other, similar, function forms. In addition, the parameters of the hyperbola varied systematically with the quantitative, and some of the qualitative, properties of the model in ways that were consistent with findings from biological organisms. These results suggest that the material events responsible for an organism's responding on RI schedules are computationally equivalent to Darwinian selection by consequences. They also suggest that the computational model developed here is worth pursuing further as a possible dynamic account of behavior. PMID- 15357513 TI - Words and our way with them. PMID- 15357514 TI - Mechanism of action of cannabinoids: how it may lead to treatment of cachexia, emesis, and pain. AB - Many patients with life-threatening diseases such as cancer experience severe symptoms that compromise their health status and deny them quality of life. Patients with cancer often experience cachexia, pain, and depression,which translate into an unacceptable quality of life. The discovery of the endocannabinoid system has led to a renewed interest in the use of cannabinoids for the management of nausea, vomiting, and weight loss arising either from cancer or the agents used to treat cancer. The endocannabinoid system has been found to be a key modulator of systems involved in pain perception, emesis, and reward pathways. As such, it represents a target for development of new medications for controlling the symptoms associated with cancer. Although the cannabinoid receptor agonist tetrahydrocannabinol and one of its analogs are currently the only agents approved for clinical use, efforts are under way to devise other strategies for activating the endocannabinoid system for therapeutic uses. PMID- 15357515 TI - Earlier initiation of treatment recommended in using erythropoietic agents for chemotherapy-induced anemia. PMID- 15357516 TI - NCCN makes changes to its guidelines for treating fever and neutropenia. PMID- 15357517 TI - Pathophysiology and palliation of inoperable bowel obstruction in patients with ovarian cancer. AB - Malignant bowel obstruction is the cause of death in the majority of women who die of ovarian cancer. Some patients are considered acceptable surgical candidates for relief of the obstruction. For many patients, however, lack of such surgical options has spawned a broad range of medical interventions, including palliative strategies to target pain and nausea and vomiting. This review discusses the general approach to patients with ovarian cancer and inoperable malignant bowel obstruction, with an emphasis on such palliative strategies. PMID- 15357518 TI - Minimally invasive treatments for spinal metastases: vertebroplasty, kyphoplasty, and radiofrequency ablation. AB - Bony metastases are the most common tumors affecting the musculoskeletal system. The most common site of such metastases is the spinal column. Many patients with these tumors present with pain and pathologic fractures. Three relatively new, minimally invasive treatments may be used to provide supportive care in this patient population: vertebroplasty, kyphoplasty, and radiofrequency ablation. In this report, we review the literature for each of these procedures and discuss their indications, results, and complications. PMID- 15357519 TI - How to use octreotide for malignant bowel obstruction. PMID- 15357520 TI - NCCN antiemesis guidelines emphasize 'delayed' emesis, new 5-HT3 inhibitors, and NK-1 blockers. PMID- 15357521 TI - Physical activity and health in the European Union. PMID- 15357522 TI - Physical activity and health: getting to know more about the European situation. PMID- 15357523 TI - Biomass fuel makes lungs a decade older--time to take action. PMID- 15357524 TI - Self-rated health and physical activity in the European Union. AB - OBJECTIVES: This article is the first in a series of four that present data about physical activity in the 15 member states of the European Union collected by the Eurobarometer 58.2. On a descriptive and multivariate level, the analysis investigates the relationship between physical activity and self-rated health in the different nations. METHODS: Data were collected in 2002 as part of the Eurobarometer by face-to-face interviews. A total of 16,230 respondents age 15 years and older were interviewed. Sample sizes ranged about 1000 respondents in most nations. Physical activity status (insufficiently active, sufficiently active, and highly active) was assessed using the last 7-days short-version of the International Physical Activity Questionnaire (IPAQ). RESULTS: On a descriptive level of analysis, results indicated positive relationships between physical activity status and self-rated health across populations subgroups as divided by age, gender, income, and educational attainment. Also on a multivariate level of analysis, physical activity status was significantly related to a better self-rated health. In an analysis on the national level, some variations in the predictive power of physical activity status for self-rated health could be observed. CONCLUSION: Results provide some indication for a positive relationship between physical activity, as assessed with the IPAQ, and self-rated health. That in most nations sufficient levels of physical activity were not positively related to self-rated health might be explained by difficulties in assessing moderate forms of physical activity, and also differences of the context (at home, for leisure, at work, for transportation) where physical activity takes place. PMID- 15357525 TI - Prevalence of overweight and obesity in Austrian male and female farmers. AB - OBJECTIVES: Risk factors for chronic and degenerative diseases, especially overweight and obesity are rarely examined among farmers. Objective of the study was to determine subjective health and health-related lifestyle among Austrian farmers. METHODS: The study was performed in 1999/2000 as a nationwide survey by mail among all Austrian farmers, men and women, all insured by the Health Insurance Agency of Farmers. RESULTS: 15.2% of the farmers were obese. 42.9% were overweight. The prevalence of overweight and obesity was high amongst 15-19 year olds, and reached a second peak among the age groups 50-59 and 60-69 years old. Geographically, the highest concentration of overweight and obese farmers was found in the eastern, flat regions of Austria and the lowest concentration in the western, mountainous areas of Austria. CONCLUSIONS: Prevalence of overweight and obesity is very high among Austrian farmers, especially when compared to the general population. The results of this study emphasize the need for further work, with respect to the development of prevention strategies to reduce the prevalence of overweight and obesity among Austrian farmers. PMID- 15357526 TI - The association of biomass fuel combustion on pulmonary function tests in the adult population of Mid-Anatolia. AB - OBJECTIVES: To assess the association of biomass as domestic energy source on respiratory function in rural areas of Mid-Anatolia in Turkey where biomass use is frequent. METHODS: In a cross-sectional study, pulmonary functions measurements of 112 cow-dung users and 153 modern energy source users, all non smokers, were assessed and compared. Several sociodemographic factors were assessed by questionnaire, and ventilatory function measurements included forced expiratory volume in one second (FEV1), forced vital capacity (FVC), FEV1/FVC ratio and the flow between the first 25 and 75 of forced expiratory flow (FEF(25 75)), and were compared, deploying univariate and multivariate methods, between the two groups. RESULTS: The individuals in the biomass exposed and unexposed group were similar regarding demographic characteristics: 67.0% were female (exposed) vs 60.1% (unexposed) with an age range of 17 to 64 (exposed) and 18 to 70 years (unexposed), and with 36% of biomass users having had pulmonary tract infections (20% in non-users). For all pulmonary function test parameters FEV1, FVC, FEV1/FVC, and FEF(25-75) a highly significant (p < 0.0001) reduction was observed in biomass users. A 12.4% (95% confidence interval: 7.0% to 17.7%) reduction in forced vital capacity was observed in multivariate linear regression. CONCLUSIONS: It is well known that biomass combustion is a relevant public health problem. The substantial differences in pulmonary functions associated with biomass use as an energy source observed in this cross-sectional study in non-smokers support that also in rural Turkey measures may be in need to reduce this public health problem. PMID- 15357527 TI - Patient willingness to pay for preventive measures in primary care: a study of five GPs in a German community. AB - OBJECTIVE: To explore the willingness of patients in a usual primary care setting to pay out-of-pocket fees for their own health promotion, in correlation with risk factors and net income, and compared to patients of an educational programme. METHODS: A standardised health survey carried out in five general practices (GPs) of a small community with a special GP-based health education programme was combined with a questionnaire to explore the special attitudes of patients from a practice sample (n = 973) and from educational courses (n = 202): covering, in addition to cardiovascular risk factors, the sociodemographic factors, net income, and out-of-pocket fees that could be spent for own health promotion. RESULTS: After attending an educational programme, the patient's willingness to spend 15-40 euros/month for their own health promotion was high but there was no correlation with the income (p < 0.56), in contradiction to the patients of the practice sample who would pay more money the more they earn (p < 0.001). High levels of cardiovascular risk were associated with low education (p < 0.001), but net income and willingness to pay for preventive measures did not significantly correlate with cardiovascular risk factors. CONCLUSION: Participants of educational courses are willing to pay a rational out-of-pocket fee for preventive measures without correlation with their incomes, thus reducing the social gradient; future preventive measures should take into account that reasonable cost sharing is well accepted by well-informed patients. PMID- 15357528 TI - [The use of guidelines in the primary care management of hypertension and diabetes]. AB - OBJECTIVES: The paper examines to what degree German primary care physicians know and work along the rules established in guidelines for arterial hypertension and diabetes mellitus. METHODS: HYDRA is a 2-stage cross-sectional point prevalence study with 1912 participating primary care settings throughout Germany including 45,125 patients. A pre-study questionnaire to assess doctors practice patterns was used. Subsequently doctors completed a structured clinical appraisal with a diagnostic workup and characterization of the current treatments provided. All patients filled out a questionnaire. RESULTS: Pre-study results show that only every second primary care physician manages the patients according to established guidelines. Further, physicians estimated their own work as problematic and not always successful. Guideline-oriented doctors were more likely to report hypertension and diabetes treatment patterns that also match guideline's recommendations. CONCLUSIONS: The study shows a considerable degree of dissatisfaction with the quality of their work among primary care physicians. Guideline-oriented doctors however reveal more frequently formally adequate management characteristics. This encouraging aspect suggests the need of more successful implementation of medical guidelines in order to achieve improved evidence-based medicine and better patient-oriented health care. PMID- 15357529 TI - Investing in surveillance: a fundamental tool of public health. AB - OBJECTIVES: The WHO Global InfoBase assembles country-level chronic disease risk factor prevalence data from WHO's member states. METHODS: The focus of this report is recent, nationally representative data. The risk factors of choice are those that make the greatest contribution to mortality and morbidity from chronic disease, can be changed through primary intervention and are easily measured in populations. RESULTS: Eight risk factors fit these criteria. They are: tobacco and alcohol use, patterns of physical inactivity, low fruit/vegetable intake, obesity, blood pressure, cholesterol and diabetes. Important to the data collection is the need to display prevalence data for these eight risk factors by age group(s) and sex and with some measure of the uncertainty of the estimate. CONCLUSIONS: This tool can be used by countries to evaluate the quality of the data that they have for chronic disease surveillance. The aim is to improve risk factor data quality and to standardize data, either through common survey instruments or by using existing country data to model risk factor estimates. These "harmonized" estimates will allow for comparisons over time and between countries. PMID- 15357530 TI - [Is methadone maintenance provided by private practitioners effective?]. AB - OBJECTIVES: Involvement of private practitioners in methadone maintenance treatment (MMT) enhances its availability. However, effectiveness of such treatments is still debated. METHODS: A retrospective case register analysis comparing the retention of private practices with that of specialised institutions. All methadone maintenance treatment starting between January 1, 1997 and December 31, 1999. Kaplan-Meier survival estimates to test for different retention times; Cox-regression procedure to control for baseline differences between the two populations. RESULTS: Even after controlling for distinct patient characteristics, MMT by private practitioners resulted in a longer retention time. CONCLUSION: Our findings favour the involvement of private practitioners. Providing better professional and financial support may enhance their participation. PMID- 15357531 TI - Prevalence of physical activity in the European Union. AB - OBJECTIVES: This article is the second in a series of four that present data about physical activity in the 15 member states of the European Union collected by the Eurobarometer 58.2. The focus of this article is on days of vigorous and moderate physical activity, days of walking, and metabolic equivalence estimates (METs) for total physical activity from 15 member states of the European Union using the International Physical Activity Questionnaire (IPAQ). METHODS: Data were collected in 2002 as part of the Eurobarometer by face-to-face interviews. A total of 16,230 respondents age 15 years and older were interviewed. Sample sizes ranged about 1000 respondents in most nations. Physical activity was assessed with the last 7-days short version of the IPAQ. RESULTS: Median METs estimates in hours per week were the highest in The Netherlands (39.43 MET-hours/week), Germany (34.65 MET-hours/week in the eastern part, 33.90 MET-hours/week in the western part), and Luxembourg (31.55 MET-hours/week). The lowest METs estimates were reported in Northern Ireland (11.55 MET-hours/week), Sweden (18.65 MET hours/week) and France (19.55 MET-hours/week). CONCLUSION: A comparison of the results with existing data on physical activity prevalence in the member states indicate some inconsistencies between studies which may be related to measurement problems, as well as to conceptual differences in the assessment of physical activity. PMID- 15357532 TI - A novel method of generating individual communications for participants in large surveys. PMID- 15357533 TI - Glaucoma arrives on managed care's doorstep. AB - Popular new prostaglandins, prostamides, and alpha2 agonists have brought about higher utilization, while their expense has produced a new cost driver in managed care organizations. The recent appearance of competitive products within these drug classes has offered patients, health plans, and physicians new treatment options. PMID- 15357534 TI - Glaucoma: prevalence, utilization, and economic implications. PMID- 15357535 TI - Clinical guidelines for the treatment of glaucoma. AB - Several documents guide screening and detection of glaucoma. The AAO guidelines are considered the current standard. The VA has established the need for regular screening based on identified factors. Finally, a task force working with NCQA to develop a glaucoma-related HEDIS measure may provide future direction in populationwide care. PMID- 15357536 TI - Glaucoma medications: a drug-therapy review. AB - Today, clinicians have unprecedented versatility in the treatment of glaucoma. Early detection, prevention, and control are promises inherent in the expansion of the pharmacological glaucoma armamentarium. Choosing the optimal regimen hinges on knowledge about the capabilities and the limitations of currently available medications. PMID- 15357537 TI - Considerations in the pharmacoeconomics of glaucoma. AB - Pharmacoeconomic research identifies, measures, and compares the costs and consequences of pharmaceutical product and service utilization. Providing optimal glaucoma care requires planning, based not only on the historically relevant factors of drug safety and efficacy, but also on data relating to the value of medical treatments. PMID- 15357538 TI - Current formulary status of glaucoma agents. PMID- 15357539 TI - Constructing disease management programs for glaucoma. AB - Glaucoma, which is associated with diabetes, hypertension, and hypothyroidism, is considered to be a good candidate for disease management. This article explains to clinicians and administrators who are interested in the identification and treatment of glaucoma what they should know about disease management and how they might benefit from establishing a DM program for glaucoma. PMID- 15357540 TI - Improving adherence to drug-treatment regimens for glaucoma. AB - Compliance with glaucoma therapy is relatively poor. Because glaucoma progresses slowly, long-term adherence is crucial to improving clinical and financial outcomes of treatment. Physicians must consider several factors before steps can be taken to promote correct use of medications. PMID- 15357541 TI - Survey of P&T committee members: glaucoma agents gaining notice. PMID- 15357543 TI - Sorry, no vacancies. PMID- 15357545 TI - A potent cocktail. PMID- 15357546 TI - In at the birth. Interview by Thelma Agnew. PMID- 15357547 TI - Making of a midwife. PMID- 15357548 TI - On the receiving end. PMID- 15357549 TI - Heat of the night. PMID- 15357550 TI - Lend a friendly ear. PMID- 15357551 TI - Benefits of pre-operative information programmes. AB - BACKGROUND: Thousands of patients undergo surgical procedures daily. Research has shown the benefits of giving pre-operative information to patients, which include decreased length of stay, less demand for analgesia post-operatively and increased patient satisfaction. However, despite this evidence, there are still many facilities with no formal policy or programme for giving pre-operative information. CONCLUSION: Nurses and managers should be made aware of the benefits and potential financial savings of pre-operative information programmes. Once education takes place, a concerted multidisciplinary effort should be made to implement a programme. This will help to ensure that patients no longer arrive at the operating theatre frightened and unaware of what will happen to them. PMID- 15357552 TI - Systematic observation of hand decontamination. AB - Meticulous infection control precautions are required to prevent healthcare associated infection, with particular attention to hand hygiene and universal precautions. A detailed observation study undertaken in the early 1990s still has clear messages for infection control today. To prevent the spread of infection in clinical settings, nurses need adequate time and resources to comply with infection control protocols. Pressure of work and staff shortages still mitigate against safe practice today despite greater awareness of the importance of infection control. PMID- 15357554 TI - Little wonders. PMID- 15357553 TI - Encouraging reflection and critical thinking in practice. AB - For reflection to become a transferable skill that is used in practice, practitioners need to learn how to combine this skill with critical thinking. This article provides practical guidance to mentors, clinical supervisors and preceptors on how this might be encouraged. PMID- 15357555 TI - No bars to learning. PMID- 15357556 TI - Avian flu and influenza pandemics in human populations. PMID- 15357557 TI - Liver glycogenoses: are they a possible cause of polyneuropathy? A cross sectional study. AB - We encountered two children suffering from liver glycogenoses (GSD) over a period of 5 years (1992-1997) who presented with a demyelinating peripheral neuropathy diagnosed by electromyography (EMG) and nerve conduction studies (NCV). The aim of the study was to evaluate the involvement of muscle and motor nerve in children suffering from liver glycogenoses. In a cross-sectional study, 22 children suffering from liver GSD (with no current neurological symptoms) and 20 age- and sex- matched clinically free children (control group) underwent creatine phospho-kinase (CPK), EMG, and NCV studies. Abnormal EMG and/or NCV studies were found in 11 children. Six (27.27 per cent) were found to have axonopathy, three (13.63 per cent) demyelinating polyneuropathy, and two (9.1 per cent) had mixed axonal and demyelinating neuropathy. Two children with axonopathy had GSD type VI, another had GSD type IV, and three had GSD of undiagnosed type. Three of those having a demyelinating polyneuropathy had GSD type III, another had GSD type IV, and the last had GSD of undiagnosed type. None were found to have a cardiomyopathy or a myopathy on EMG. This is the first report of neuropathy associated with GSD types III, IV, and VI in children. It might be discovered by EMG and/or NCV studies in a clinically, neurologically normal child suffering from GSD, or present as an acute polyneuropathy. PMID- 15357558 TI - Use of volunteer medical brigades to assess growth in Honduras. AB - We endeavored to determine whether a visiting volunteer medical group could effectively measure growth status among children in a developing country, identify predictors of poor growth, and thus participate in nutritional surveillance. Cross-sectional measurements of growth and diagnosis of current clinical conditions were made. A sample of 3284 Honduran children aged 2-11 years who sought care from volunteer medical brigades between January 2000 and May 2001 were included in the study. Main outcome measures were height-, weight- and body mass index-for-age z scores. Compared with standard reference data, 10 per cent of children were moderately underweight and 3.3 per cent severely underweight, while 13.7 per cent were moderately stunted and 6.4 per cent were severely stunted. After simultaneous adjustment for demographics, clinical conditions, and village characteristics, nutritional complaints were associated with lower body mass index- and weight-for-age, while children seen in villages with clean water and higher levels of development had higher body mass index- and weight-for-age. Older children had poorer growth for all parameters. It was concluded that Honduran children seeking care from a volunteer medical brigade were underweight and stunted compared with a reference population. This study demonstrates that a visiting volunteer group can collect quality growth data that may assist in nutritional surveillance, identify predictors of poor growth, and provide information useful for local public health initiatives. PMID- 15357559 TI - Comparison of pulmonary function between children living in rural and urban areas in northern Nigeria. AB - Children in northern Nigeria and elsewhere in the hot, arid western Sahel, are at risk of having their lung function compromised by a variety of factors, including undernutrition, environmental factors (e.g. airborne pollutants such as dust and smoke from wood fires), chronic upper-respiratory tract infections, and low socioeconomic class. We were interested in using spirometry to compare the pulmonary function of Nigerian children and adolescents aged 6-18 years who were living in urban and rural settings with the corresponding standards for African American children. A total of 183 boys and girls in the rural village of Sabon Fobur on the Jos Plateau and another 128 boys and girls in the city of Jos were tested to determine their forced vital capacity (FVC), FVC at 1 s (FVC1), and peak expiratory flow (PEF). The nutritional status of the subjects was determined by measuring the body mass index (BMI), triceps skin-fold thickness, and mid-arm circumference, and fat-free mass (FFM) and fat mass (FM) by bioelectrical impedance analysis. According to the results of anthropometry, the subjects in Sabon Fobur and Jos were lean but generally adequately nourished. The mean FVC, FVC1 and PEF values for the rural males were 1.851,1.761, and 3.521, and for the urban males they were 1.971,1.791, and 3.471, respectively. The corresponding values for the rural females were 1.791,1.701, and 3.371, and for the urban females they were 1.761,1.671, and 3.091. These values were approximately 100 per cent of the corresponding values for African-American children. In general, strong correlations were found between each of the three lung function parameters and age, weight, height (only for the males), BMI, MAC, and FFM. These results show that: (1) the lung function of Nigerian children and adolescents living in either rural or urban areas were similar and compared favorably with African American standards, and (2) weight was as important as height in determining pulmonary function. The inclusion of FFM as an explanatory variable did notfurther increase the accuracy of the prediction, even in a population where malnutrition may be prevalent. Therefore, we conclude that measurements of height and weight are all that are required for the assessment of lung function using spirometry in Nigerian children. PMID- 15357560 TI - Environmental regulators of biological variation. PMID- 15357561 TI - Use of magnesium sulphate in severe persistent pulmonary hypertension of the newborn. AB - Persistent pulmonary hypertension in the newborn (PPHN) is a neonatal emergency due to the failure to achieve normal cardiopulmonary adaptation following delivery. A clinical trial to assess the efficacy of magnesium sulphate (MgSO4) in the treatment of PPHN was conducted in the tertiary level neonatal intensive care unit of RIPAS Hospital, Brunei Darussalam over a period of 2 years. Twelve newborn babies admitted consecutively to the neonatal intensive care unit with severe hypoxemia due to persistent pulmonary hypertension were included in this trial. A loading dose of 200 mg/kg MgSO4 was given over a period of 20 min, followed by a continuous infusion at the rate of 20-150 mg/kg/h to obtain a serum magnesium level between 3.5 and 5.5 mmol/l. No other vasodilators were used and babies were not hyperventilated. Dopamine was commenced at 5-10 microg/kg/min before the loading dose of MgSO4 was given. Mean blood pressure was maintained with short periods of dopamine alone or in combination with dobutamine. Oxygen index (OI) and alveolar-arterial oxygen gradient (A-aDO2) showed significant improvement within 24 h of treatment. In this study, MgSO4 was found to be a safe and effective pulmonary vasodilator. MgSO4 could be used as a first-line vasodilator in developing countries because of its low cost and high efficacy. Parenteral MgSO4 is easy to administer and monitor during treatment. When MgSO4 is used, systemic hypotension can be adequately controlled with ionotropes. PMID- 15357562 TI - Ketamine and midazolam for invasive procedures in children with malignancy: a comparison of routes of intravenous, oral, and rectal administration. AB - We investigated the efficacy of a combination of ketamine and midazolam, comparing intravenous, oral, and rectal administrations for invasive procedures in children with malignancy. Seventy-three children under 5 years of age, who were scheduled for invasive procedure, were assigned to one of three groups: IV group (n = 25), ketamine 1 mg/kg and midazolam 0.05-0.1 mg/kg were given intravenously; PO group (n = 24), ketamine 3 mg/kg and midazolam 0.5 mg/kg were given orally; and PR group (n = 24), ketamine 3 mg/kg and midazolam 0.5 mg/kg given rectally. Vital signs including blood pressure, pulse rate, respiratory rate, and oxygen saturation were monitored, and patients were observed for side effects. Optimal sedation (drowsy and asleep) was provided in 78 per cent of all patients and no statistical difference was observed among the three groups. No severe complications were observed in all groups. Recovery time from sedation was significantly longer in the intravenous group (>120 min in two patients). Hallucination was noted in three (12 per cent) patients given intravenous medication, but not in those given oral or rectal medications. It is concluded that intravenous, oral, and rectal midazolam/ketamine are equally effective for invasive procedures in children with malignancy. The use of intravenous ketamine/midazolam may produce prolonged sedation and psychedelic effects in children. These adverse effects may alter the child's comfort and parental satisfaction. PMID- 15357563 TI - Microbiological identification in cystic fibrosis patients with CFTR I1234V mutation. AB - Recurrent and chronic bacterial pulmonary infection is the major cause of morbidity and mortality in cystic fibrosis (CF). Over 6 months, 72 sputa or oropharyngeal samples were examined from 36 Arab Bedouin CF patients attending Hamad General Hospital, Doha, Qatar. More than 100 pathogens were isolated, mostly Haemophilus influenzae, Staphylococcus aureus and Pseudomonas aeruginosa. Unusual pathogens included Stenotrophomonas maltophilia, Acaligenes xylosoxidans and Mycobacterium abscessus. It is concluded that microbiological biodiversity in the lower airways of CF patients continues to be underestimated and that CF patients harbouring mucoid strains of P. aeruginosa are at a higher risk of acquiring more unusual organisms and probably have a worse prognosis. PMID- 15357564 TI - The probiotic effect of Saccharomyces boulardii in a pediatric age group. AB - The aim of this study was to determine the efficacy of S. boulardii in diarrhea associated with commonly used antibiotics such as sulbactam-ampicillin (SAM) and azithromycin (AZT). Four hundred and sixty-six patients were assigned to four different groups as follows: group 1:117 patients receiving SAM alone; group 2:117 patients receiving SAM and S. boulardii, group 3:105 patients receiving AZT alone; group 4:127 patients receiving AZT and S. boulardii. Antibiotic-associated diarrhea was seen in 42 of the 222 patients (18.9 per cent) receiving an antibiotic without the probiotic, and in 14 of the 244 patients (5.7 per cent) who received both the probiotic and the antibiotic (p < 0.05). In the group receiving SAM where S. boulardii use was found to be significant, the use of S. boulardii decreased the diarrhea rate from 32.3 to 11.4 per cent in the 1-5 years age group (p < 0.05). This is a pioneering study investigating combined antibiotic and probiotic use in pediatric diarrhea patients. PMID- 15357565 TI - Mother-to-Infant transmission of hepatitis C virus (HCV) in Brazil. AB - Sixty-one women with anti-HCV antibodies, detected by a third-generation enzyme immunoassay (EIA3), were prospectively recruited for investigation of vertical HCV transmission during child-birth, at the University Hospital of the Catholic University of Campinas, Brazil, between January 1994 and July 1998. Six of the women presented coinfection with the human immunodeficiency virus type 1 (HIV-1). All of the 72 children born in this period were followed at least until they were 18 months of age. Analyses of anti-HCV, HCV RNA, and alanine aminotransferase were performed in a minimum of two blood samples during follow-up. One (2.4 per cent; 95 per cent CI, 2.2-7) of the 42 children born to HCV viremic mothers was both anti-HCV and HCV RNA-positive, with altered ALT levels. Passively transferred maternal anti-HCV antibodies became undetectable within 9-12 months. None of the nine infants born to HIV-1 infected mothers were infected either by HIV or HCV. Thus, the mother-infant HCV transmission rate is low and seems to be associated with maternal HCV RNA positivity. PMID- 15357566 TI - A novel missense mutation A1081P in the cystic fibrosis transmembrane conductance regulator (CFTR) gene identified in a Laotian patient with congenital bilateral absence of the vas deferens. AB - Cystic fibrosis is the most common autosomal disorder in the Caucasion population. However, the disease is rare in Asia and little is known about the spectrum of CF transmembrane conductance regulator, CFTR, mutations in this population. We studied a 39-year-old Loatian patient with congenital bilateral absence of the vas deferens and identified a novel missense mutation in exon 17b (3373G>C). Identification of novel mutations in this Asian population is of particular interest when designing a genetic testing strategy in Asian countries and also in other countries where immigration from Asia is common. PMID- 15357567 TI - Thrombocytopenia in hepatitis A--an atypical presentation. AB - Acute hepatitis due to hepatitis A virus (HAV) is usually a benign self-limiting disease during childhood. Autoimmune manifestations are rare with acute HAV, especially in children. We report an 8-year-old boy with severe thrombocytopenia as an initial manifestation of acute HAV infection. Bone marrow examination suggested peripheral destruction of platelets. His clinical course was complicated by persistent severe thrombocytopenia and progressive cholestasis requiring prednisolone therapy. PMID- 15357568 TI - Pseudo-Bartter's syndrome in an Egyptian infant with cystic fibrosis mutation N1303K. AB - An Egyptian infant with the common CFTR mutation N1303K in exon 21 developed alkalosis, electrolyte disturbance, and pancreas insufficiency. We emphasized the need to consider the possibility of cystic fibrosis (CF) in the Arab world. The frequency of N1303K mutation in the Middle East and its distribution are both reviewed. PMID- 15357569 TI - Current status of urinary iodine excretion levels in 116 districts of India. PMID- 15357570 TI - Immunohistochemistry of tuberculin skin test. PMID- 15357571 TI - Postpneumonic empyema in childhood: a little goes a long way. PMID- 15357572 TI - Colloquium report: improving the health of school age children in an era of HIV/AIDS in Durban, South Africa. AB - Increased morbidity and mortality due to the HIV/AIDS epidemic in South Africa necessitates the examination of policies for the protection and support of school age children. At a colloquium held in Durban participants from government and non government health and education sectors briefed delegates on key policies and programmes for promoting the health and mental health of school age children. Researchers gave evaluative accounts of the impacts of these measures and presented an overview of South African children's health and mental health needs. In the discussions that followed four critical issues emerged: intersectoral collaboration, accountability, retraining middle management, and a more comprehensive strategy to support pupils and teachers infected and affected by HIV/AIDS. PMID- 15357573 TI - Social self-sorting in aqueous solution. AB - Self-sorting-the ability to efficiently distinguish between self and nonself-is common in nature but is still relatively rare in synthetic supramolecular systems. We report a 12-component mixture comprising 1-11 and KCl that undergoes thermodynamically controlled self-sorting in aqueous solution based on metal ligand, ion-dipole, electrostatic, charge-transfer interactions, as well as the hydrophobic effect. We refer to this molecular ensemble-characterized by high fidelity host-guest interactions between components-as a social self-sorting system to distinguish it from narcissistic self-sorting systems based on self association processes. The influence of several key variables-temperature, pH, concentration, and host/guest stoichiometry-was explored by a combination of simulation and experiment. Variable temperature NMR experiments, for example, revealed a kinetically controlled irreversible process upon cycling from 298 to 338 K, which is an emergent property of this molecular ensemble. Variable pH and concentration experiments, in contrast, did not reveal any emergent properties of the molecular ensemble. Simulations of a four-component mixture establish that by proper control of the relative magnitude of the various equilibrium constants, it is possible to prepare socially self-sorted mixtures that are responsive (irresponsive) to host/guest stoichiometry over narrow (broad) ranges. The 12 component mixture is relatively irresponsive to host/guest stoichiometry. Such social self-sorting systems, like their natural counterparts, have potential applications as chemical sensors, as artificial regulatory elements, and in the preparation of biomimetic systems. PMID- 15357574 TI - Structural modifications of plumieride isolated from Plumeria bicolor and the effect of these modifications on in vitro anticancer activity. AB - Plumieride was isolated as one of the major components from the biologically active methanolic extract of the bark of Plumeria bicolor (family Apocynaceae). For investigating the effect of substituents on cytotoxic activity it was modified into a series of compounds. Replacing the methyl ester functionality of plumieride with alkyl amides of variable carbon units improved the cytotoxic activity, and a correlation between overall lipophilicity and cytotoxic activity was observed. In plumieride, the glucose moiety was converted into a di- and trisaccharide by following the protection and deprotection approach, and the resulting compounds produced enhanced cytotoxicity. However, these compounds were found to be less effective than plumeiride containing a dodecyl (12 carbon units) amide group. Among all of the derivatives, the naturally occurring plumieride showed the least cytotoxicity (50% cell kill = 49.5 microg/mL), and the dodecyl amide analogue of plumieridepentaacetate produced the best efficacy (50% cell kill = 11.8 microg/mL). The di- and trisaccharide analogues were found to be slightly less effective than the dodecyl derivative (50% cell kill = 15-17 microg/mL). The in vitro cytotoxicity of the plumieride analogues was determined in radiation-induced fibrosarcoma (RIF) tumor cells. PMID- 15357575 TI - Proton as the simplest of all catalysts for [2 + 2] cycloadditions: DFT study of acid-catalyzed imine metathesis. AB - The mechanism of imine metathesis was studied as a prototype reaction for the impact that heteroatom substitution has on thermally forbidden [2 + 2] addition reactions using high-level density functional theory in combination with a continuum solvation model. The intuitively expected high activation barriers were confirmed for N-alkyl- and N-aryl-substituted imine reactants with transition state free energies of 78.8 and 68.5 kcal/mol, respectively, in benzene. The computed reaction energy profiles were analyzed to discover possible strategies for lowering the transition state energy. Protonation of the imine nitrogen was proposed as a possible catalytic route and was explicitly modeled. The computed reaction energy profile shows that protonation of one of the imine reactants has an enormous effect on the overall rate of metathesis and lowers the activation barrier by as much as 37.3 and 30.6 kcal/mol for the N-alkyl and N-aryl reactants, respectively. These results suggest that acid-catalyzed imine metathesis should be amenable at elevated temperatures. Furthermore, the protonation of both reactants of the metathesis reaction is predicted to be not productive owing to electrostatic repulsion of the reactants, thus suggesting that there should be an optimum pH for the catalytic turnover. A detailed analysis of the catalytic mechanism is presented, and the primary driving force for the catalysis is identified. Upon protonation of the imine nitrogen, the key [2 + 2]-addition step becomes asynchronous and one of the two intermolecular N-C bonds is formed before traversing the transition state, resulting in a substantial net decrease of the overall energy requirement. The general applicability of this intuitively understandable mechanism for designing structural features for lowering the energy of transition state structures is explored. PMID- 15357576 TI - Potent inhibitors of the hepatitis C virus NS3 protease: design and synthesis of macrocyclic substrate-based beta-strand mimics. AB - The virally encoded NS3 protease is essential to the life cycle of the hepatitis C virus (HCV), an important human pathogen causing chronic hepatitis, cirrhosis of the liver, and hepatocellular carcinoma. The design and synthesis of 15 membered ring beta-strand mimics which are capable of inhibiting the interactions between the HCV NS3 protease enzyme and its polyprotein substrate will be described. The binding interactions between a macrocyclic ligand and the enzyme were explored by NMR and molecular dynamics, and a model of the ligand/enzyme complex was developed. PMID- 15357577 TI - Modeling substituent and conformational effects on the reactivity of antitumor agents containing a cyclopropylcyclohexadienone subunit. AB - The uncatalyzed alkylation reactions of ammonia by the parent spirocyclopropylcyclohexadienone (6), its 3-amino analogue (7), the cyclic derivative (8), its N-formyl derivative (9), and a closer model (10) of the CPI (1-4) drugs have been investigated in gas phase and in water solvent bulk, using density functional theory at the B3LYP level with several basis sets and the C PCM solvation model. The effect of several structural key features such as the vinylogous amide conjugation, the acylation of the 2-amino substituent, the ring constraint of the heterocyclic nitrogen atom at C(2) carbon in a ring, and the presence of a condensed pyrrole ring on the reaction activation energy have been investigated. Substrate 7, which is a flexible conformational model of the cyclopropylpyrroloindole moiety (CPI) contained in the duocarmycins, has been used to model the shape-dependent reactivity of these drugs, in gas phase and water solutions. The calculations indicate that shape dependence of reactivity is strongly operative both in gas phase and in polar solvents, since conformational effects are capable of reducing the reaction activation energy by -8.4 and -4.3 kcal mol(-1) in gas phase and in water solution, respectively, that is required to promote "conformational catalysis". PMID- 15357578 TI - Delta-peptides and delta-amino acids as tools for peptide structure design--a theoretical study. AB - An overview on all possible helix types in oligomers of delta-amino acids (delta peptides) and their stabilities is given on the basis of a systematic conformational analysis employing various methods of ab initio MO theory (HF/6 31G*, B3LYP/6-31G*, PCM//HF/6-31G*). A wide variety of novel helical structures with hydrogen-bonded pseudocycles of different size are predicted. Since a delta amino acid constituent may replace a dipeptide unit in alpha-peptides, there are close relationships between the secondary structures of peptides with delta-amino acid residues and typical secondary structures of alpha-peptides. However, the preference of gauche conformations at the central C(beta)-C(gamma) bonds of delta amino acids, which correspond to the peptide linkages in alpha-peptides, over staggered ones makes completely novel structure alternatives for helices and turns more probable. The peculiarities of beta-turn formation by sugar amino acids derived from delta-amino acids are compared with the turn formation in delta-amino acid residues and in alpha-peptides. The considerable potential of secondary structure formation in delta-peptides and single delta-amino acid constituents predicted by ab initio MO theory may stimulate experimental work in the field of peptide and foldamer design. PMID- 15357579 TI - Hydrogen-bonding-induced planar, rigid, and zigzag oligoanthranilamides. Synthesis, characterization, and self-assembly of a metallocyclophane. AB - Four oligoanthranilamides 1-4, which are incorporated with three, five, seven, or nine benzene units, respectively, have been synthesized and characterized. X-ray analysis, 1D and 2D (1)H NMR, and IR experiments reveal that all the new oligoamides adopt rigid, planar and zigzagged conformations in both solution and solid state, which are stabilized by intramolecular three-center hydrogen bonding. A 5-mer oligomer 22, which is incorporated with two acetylene groups at the ends, has also been synthesized and utilized for the self-assembly of a rigid hydrogen-bonded metallocyclophane. The new rigid oligoanthranilamides represent useful building blocks for the construction of supramolecular architectures. PMID- 15357580 TI - Solvophobically-driven oligo(ethylene glycol) helical foldamers. Synthesis, characterization, and complexation with ethane-1,2-diaminium. AB - Oligo(ethylene glycols) 1a-h, which are incorporated with one to eight 2,3 naphthylene units, respectively, have been synthesized and characterized. The conformational changes of the new oligomers have been investigated in chloroform acetonitrile binary solvents by the UV-vis, (1)H NMR, and fluorescent spectroscopy. It has been revealed that the naphthalene units in hexamer 1f, heptamer 1g, and octamer 1h are driven by solvophobic interaction to stack in polar solvents. As a result, compact helical conformations are formed that give rise to a cavity similar to that of 18-crown-6. Shorter oligomers 1b-e exhibit weaker folding tendency. (1)H NMR studies reveal that 1f-h are able to complex ammonium or ethane-1,2-diaminium 19, but not secondary ammonium compounds. The association constants of complexes 1f.19, 1g.19, and 1h.19 in acetonitrile are determined to be 3.5(+/-0.4) x 10(3), 1.0(+/-0.12) x 10(4), and 2.5(+/-0.4) x 10(4) M(-1), respectively, with the (1)H NMR titration method. For comparison, hexamer 22, which incorporates six 1,5-naphthylene units, is also prepared. The UV-vis and fluorescent investigations show that 22 is also able to fold in polar solvents, but no helical structure can be produced due to mismatch of the stacking naphthalene units and consequently there is no obvious complexation between 22 with ethane-1,2-diaminium ion. The structures of the longest foldamer 1h and its complex with 19 have been studied with molecular mechanics calculations. This work represents a new approach to building folding conformations from flexible linear molecules. PMID- 15357581 TI - Intramolecular cyclization of ene-imine using dibutylzirconocene. AB - The reaction of ene-imine with Cp(2)ZrBu(2) was carried out. When a crude imine, which was prepared from ene-aldehyde and primary amine in the presence of MgSO(4), was treated with Cp(2)ZrBu(2) at room temperature overnight, cyclopentane derivative having trans-substituents was obtained in high yield along with a small amount of cyclopentane derivative having cis-substituents. Presumably, cis-zirconacycle is a thermodynamic product. Reactions using various ene-imines were carried out. In the case of ene-imine prepared from ene-aldehyde and (t)BuNH(2), only cyclopentane having cis-substituents was produced. In this reaction, chiral amine was used, and diastereoselective cyclization of ene-imine was carried out. As a result, cyclopentane derivative having cis-substituents was obtained in an optically active form after hydrogenolysis of the cyclized compound. PMID- 15357582 TI - Synthesis of chiral amino epoxyaziridines: useful intermediates for the preparation of chiral trisubstituted piperidines. AB - Chiral aminoalkyl epoxyaziridine 1 is synthesized in high yield and diastereoselectivity from L-serine. Ring opening of epoxyaziridine 1 with primary amines is carried out with total chemo- and regioselectivity, affording chiral polyfunctionalized piperidines 8. The structure of these trisubstituted piperidines is established by NMR studies. PMID- 15357583 TI - Interhalogens (ICl/IBr) and AgOTf in thioglycoside activation; synthesis of bislactam analogues of ganglioside GD3. AB - The novel promoter system IX/AgOTf (X = Cl or Br) has been evaluated in the synthesis of two bislactam analogues of GD3. We have carried out two high yielding galactosylations in 97% and 98% yield, respectively, using ICl/AgOTf, and four sialylations in 93%, 59%, 40%, and 44% yield, using IBr/AgOTf. The choice of interhalogen (IX) is determined by the donor type used in the glycosylation. We also report some mechanistic investigations leading to further optimization of the IX/AgOTf promoter system. PMID- 15357584 TI - Practical synthesis of a potent hepatitis C virus RNA replication inhibitor. AB - A practical, efficient synthesis of 1, a hepatitis C virus RNA replication inhibitor, is described. Starting with the inexpensive diacetone glucose, the 12 step synthesis features a novel stereoselective rearrangement to prepare the key crystalline furanose diol intermediate. This is followed by a highly selective glycosidation to couple the C-2 branched furanose epoxide with deazapurine. PMID- 15357585 TI - Solvent-dependent chemoselectivities in additions of beta-carbonyl imines to allyltrimethylsilane with CTAN. AB - The oxidative coupling of beta-carbonyl imines and allyltrimethylsilane with CTAN were investigated in CH(3)CN and CH(2)Cl(2). In CH(3)CN allylation products were obtained predominantly, while in CH(2)Cl(2), dihydropyrrole products were obtained exclusively. Solvent-assisted nucleophilic cleavage of the intermediate beta-silyl cation is proposed to play a role in the solvent-dependent chemoselectivity. PMID- 15357586 TI - Synthesis and high-throughput screening of N-acetyl-beta-hexosaminidase inhibitor libraries targeting osteoarthritis. AB - C1 Nitrogen iminocyclitols are potent inhibitors of N-acetyl-beta hexosaminidases. Given hexosaminidases' important roles in osteoarthritis, we developed two straightforward and efficient syntheses of C1 nitrogen iminocyclitols from two readily available starting materials, D-mannosamine hydrochloride and the microbial oxidation product of fructose. A diversity oriented synthetic strategy was then performed by coupling these core structures with various aldehydes, carboxylic acids, and alkynes to generate three separate libraries. High-throughput screening of the generated libraries with human N acetyl-beta-hexosaminidases produced only moderate inhibitory activities. However, the synthetic approach and screening strategy for these compounds will be applied to develop new potent inhibitors of human N-acetyl-beta hexosaminidases, particularly when combined with the structural information of these enzymes. PMID- 15357588 TI - Diastereoselective dihydroxylation and regioselective deoxygenation of dihydropyranones: a novel protocol for the stereoselective synthesis of C1-C8 and C15-C21 subunits of (+)-discodermolide. AB - Diastereoselective dihydroxylation of dihydropyranones and subsequent regioselective alpha-deoxygenation provides 1,3-trans-beta-hydroxy-delta-lactones stereoselectively. This protocol has been applied for the synthesis of C(1)-C(8) and C(15)-C(21) subunits of (+)-discodermolide. PMID- 15357587 TI - Macrocyclic bisbinaphthyl fluorophores and their acyclic analogues: signal amplification and chiral recognition. AB - A series of optically active macrocyclic and acyclic bisbinaphthyls have been synthesized and characterized. The structure of one of the bisbinaphthyl macrocycles has been established by a single-crystal X-ray analysis. The UV and fluorescence spectra of these chiral compounds in various solvents and at different concentrations are studied. Formation of excimers is observed for the macrocyclic bisbinaphthyl compounds. Introduction of conjugated substituents to the 6,6'-positions of the binaphthyl units in the macrocycles leads to greatly amplified fluorescence signals. Using the 6,6'-substituted bisbinaphthyl macrocycles in place of the unsubstituted macrocycles allows a 2 orders of magnitude reduction in the sensor concentration for the fluorescence measurements. These macrocycles have exhibited highly enantioselective fluorescent enhancements in the presence of chiral alpha-hydroxycarboxylic acids and N-protected alpha-amino acids. They are useful as fluorescent sensors for chiral recognition. The macrocycles show much greater enantioselectivity in the substrate recognition than their acyclic analogues. PMID- 15357589 TI - Highly selective synthesis of bicyclic quinolizidine alkaloids and their analogues via double RCM reaction of N-alkynyl-N-(1,omega)-alkadienyl acrylamides. AB - The double ring-closing metathesis reaction of N-alkynyl-N-(1,omega)-alkadienyl acrylamides 1 using first- or second-generation Grubbs' catalyst afforded, in a highly selectively manner, the fused bicyclic quinolizidine alkaloid derivatives and their analogues bearing a 1,3-diene moiety, which may further undergo a Diels Alder reaction with a dienophile to afford N-containing polycyclic compounds. The excellent selectivity of fused/dumbbell-mode cyclization has been realized by the higher reactivity of the electron-rich C=C bond or carbon-carbon triple bond combined with the lower reactivity of the electron-deficient C=C bond toward metallocarbenes and the thermodynamically more stable nature of fused bicyclic compounds 3 vs dumbbell-type bicyclic compounds 4. PMID- 15357590 TI - 3'-C-Branched LNA-type nucleosides locked in an N-type furanose ring conformation: synthesis, incorporation into oligodeoxynucleotides, and hybridization studies. AB - Three protected 3'-C-branched LNA-type phosphoramidite building blocks 17, 27, and 38, containing furanose rings locked in an N-type conformation, were synthesized from a known 3-C-allyl allofuranose derivative using strategies relying on the introduction of the branching alkyl chain before condensation with the nucleobase. Synthesis of 3'-C-hydroxypropyl derivatives proved superior to synthesis of the 3'-C-hydroxyethyl derivatives, and the former was converted into the corresponding 3'-C-aminopropyl derivatives. Phosphoramidites 27 and 38 were subsequently applied on an automated DNA synthesizer leading to the introduction of three novel 3'-C-branched LNA-type monomers X, Y, and Z into oligodeoxynucleotides and studies of their effect on the hybridization properties. A duplex-stabilizing effect of introducing 3'-C-aminopropyl-LNA monomer Y, relative to 3'-C-hydroxypropyl-LNA monomer X, was observed, especially at low salt conditions. This indicates that the primary amino group of monomer Y is protonated under the hybridization conditions applied and that positioning of this positively charged group in the major groove has a significant duplex stabilizing effect. Monomer Y was by an on-column conjugation method further functionalized by a glycyl unit to give monomer Z that showed a less stabilizing effect than monomer Y. PMID- 15357591 TI - Synthesis of mono- and difluoroacetyltrialkylsilanes and the corresponding enol silyl ethers. AB - A variety of mono- and difluoroacetylsilanes and the corresponding silyl enol ethers were prepared from trifluoroethanol and chlorotrialkylsilanes in the presence of LDA through retro-Brook rearrangement. Sterically demanding silyl groups, especially those bound to oxygen, resulted in higher yields of difluoroacetylsilanes. The yields of difluoroacetylsilanes were also dramatically affected by the method of the termination of the reaction. Difluorohaloacetylsilanes were prepared from the corresponding difluoroethenyl silyl ethers with electrophilic halogenating reagents in good yields. A gamma fluorinated beta-diketone 9a was prepared from monofluoroacetyltriisopropylsilane by nucleophilic acylation with methyl trifluoroacetate. PMID- 15357592 TI - A highly stereoselective synthesis of optically active trisubstituted 1,2 ethylenediamines: the first example of grignard addition to N-diphenylphosphinoyl ketimines derived from amino acids. AB - The efficient synthesis of optically active trisubstituted 1,2-ethylenediamines is described. Addition of aryl and/or alkyl Grignard reagents to alpha-amino N diphenylphosphinoyl ketimines derived from alpha-amino acids was demonstrated to afford the desired trisubstituted 1,2-ethylenediamines in good yields and with high diastereoselectivities. Subsequent removal of the diphenyphosphinoyl group from the adduct was smoothly accomplished in reasonable yield without racemization under newly developed reductive conditions. PMID- 15357593 TI - Asymmetric intramolecular michael addition of alpha-sulfinyl vinylic carbanion to enoates. AB - The first example of an asymmetric intramolecular Michael addition reaction with use of alpha-lithiated vinylic sulfoxide as a Michael donor is reported. Michael addition of the alpha-lithiated vinylic sulfoxide to (Z)-enoates proceeds with high diastereoselectivity to give the adducts with (R)-configuration at the beta position of the ester in the five-membered-ring formation. The selectivity was reversed in the six-membered-ring formation. The resulting ester enolates were reacted with alkyl halides or benzaldehyde with high diastereoselectivity. PMID- 15357594 TI - Stereoselective synthesis of allyl-C-mannosyl compounds: use of a temporary silicon connection in intramolecular allylation strategies with allylsilanes. AB - Methyl mannoside 16 containing an allyldimethylsilyl ether at C(2) was synthesized in nine steps from D-mannose. Reaction with TMSOTf in MeCN at room temperature effected C-glycosylation to provide the alpha-allyl-C-mannosyl product 18 with excellent stereoselectivity. Crossover experiments over a range of reaction concentrations proved that reaction was proceeding via an intermolecular pathway rather than the hoped-for intramolecular delivery route. The exceptionally high stereoselectivity of this allylation in the presence of an acid-scavenger, 2,6-DTBMP, can be attributed to the allylsilyl ether 16 behaving as the allylating agent. Geometrical constraints in the seven-membered ring transition state account for the lack of intramolecular allyl transfer. Attaching a modified allylsilane 29a-c to C(2)OH of methyl mannoside 15 improved matters. Reaction of the tethered mannosides 27a-c with TMSOTf in the presence of 2,6 DTBMP in MeCN at rt provided a range of products, which depended on the size of the alkyl substituents at the silyl ether tether. Diene products were the major compounds irrespective of the size of the alkyl substituents at the silyl ether tether. Their formation can be understood by intramolecular allylation of the allylsilane on to the activated anomeric center, followed by collapse of the intermediate carbocation by preferential attack of an external nucleophile at the silyl ether tether, rather than at the allylic silicon center. A cascade of further reactions rationalizes the formation of the 2-dienyl-substituted tetrahydrofuran 30 and dienes 39 and 40. The desired beta-allyl-C-mannosyl products 42 and 43 were obtained, albeit in low yield, when bulky ethyl and isopropyl groups were employed at the silyl ether tether. Stereospecific oxidative cleavage of the silyl tether in 42 and 43 provided the corresponding stereodefined diols 44 and 45, respectively. Attempts to improve the yield and diastereoselectivity of the desired beta-allyl-C-mannosyls by moving to a sulfoxide mannosyl donor, which could be activated at low temperature, proved unsuccessful. PMID- 15357595 TI - The reaction of cyclopentyne with ethene: concerted vs stepwise mechanism? AB - The cycloaddition of cyclopentyne with ethene was examined using (U)B3LYP and CASSCF methods to discern the reaction mechanism. (U)B3LYP/6-31G* and (U)B3LYP/6 311+G* slightly favor the concerted pathway, whereas CASSCF(4,4)/6-31G* and CASCF(6,6)/6-31G* favor the diradical pathway. MRMP2 using the CASSCF(4,4) wave function also favors the diradical mechanism. In the context of a diradical pathway, the experimentally observed complete retention of stereochemistry for this reaction is understood in terms of stereochemical control resulting from dynamic effects. PMID- 15357596 TI - Effect of changing electrophilic center from C=O to C=S on rates and mechanism: pyridinolyses of O-2,4-dinitrophenyl thionobenzoate and its oxygen analogue. AB - Second-order rate constants have been measured spectrophotometrically for the reactions of O-2,4-dinitrophenyl thionobenzoate (1) and 2,4-dinitrophenyl benzoate (2) with a series of substituted pyridines in 80 mol % H(2)O/20 mol % DMSO at 25.0 +/- 0.1 degrees C. The Bronsted-type plots obtained are nonlinear with beta(1) = 0.26, beta(2) = 1.07, and pK(a) degrees = 7.5 for the reactions of 1 and beta(1) = 0.40, beta(2) = 0.90, and pK(a) degrees = 9.5 for the reactions of 2, suggesting that the pyridinolyses of 1 and 2 proceed through a zwiterionic tetrahedral intermediate T(+/-) with a change in the rate-determining step at pK(a) degrees = 7.5 and 9.5, respectively. The thiono ester 1 is more reactive than its oxygen analogue 2 except for the reaction with the strongest basic pyridine studied (pK(a) = 11.30). The k(1) value is larger for the reactions of 1 than for those of 2 in the low pK(a) region, but the difference in the k(1) value becomes negligible with increasing the basicity of pyridines. On the other hand, 1 exhibits slightly larger k(2)/k(-1) ratio than 2 in the low pK(a) region but the difference in the k(2)/k(-1) ratio becomes more significant with increasing the basicity of pyridines. Pyridines are more reactive than alicyclic secondary amines of similar basicity toward 2 in the pK(a) above ca. 7.2 but less reactive in the pK(a) below ca. 7.2. The k(1) value is slightly larger, but the k(2)/k(-1) ratio is much smaller for the reactions of 2 with pyridines than with isobasic secondary amines in the low pK(a) region, which is responsible for the fact that the weakly basic pyridines are less reactive than isobasic secondary amines. PMID- 15357597 TI - Synthesis of 1,4-benzodiazepine-3,5-diones. AB - Even though benzodiazepines have a strong position in medicinal chemistry, very few synthetic routes to 1H-1,4-benzodiazepine-3,5(2H,4H)-diones have ever been published and the claimed products have often been poorly characterized. Through the present work several 1H-1,4-benzodiazepine-3,5(2H,4H)-diones have become available from N-carbamoylmethylanthranilic acids. The required ring closures were achieved only when the amino groups of the starting materials were substituted with electron withdrawing groups such as acetyl, alkyloxycarbonyl, or nitroso. During the synthetic work a novel ring contraction rearrangement from a 1-nitroso-1H-1,4-benzodiazepine-3,5(2H,4H)-dione to a 3H-quinazoline-4-one was observed. The proposed mechanism involves elimination of HNO followed by a proton mediated loss of CO. The 1-nitrosated 1,4-benzodiazepinediones could be separately denitrosated to the corresponding amino compounds. PMID- 15357598 TI - Rhodium(II)-catalyzed aziridination of allyl-substituted sulfonamides and carbamates. AB - Several unsaturated sulfonamides underwent intramolecular aziridination when treated with PhI(OAc)(2), MgO, and catalytic Rh(2)(OAc)(4) to give bicyclic aziridines in excellent yield. Treatment of the resulting azabicyclic sulfonamides in methanol in the presence of p-TsOH resulted in exclusive opening of the aziridine ring at the most substituted position affording six- and seven membered ring products in high yield. In contrast, the intramolecular aziridination of several cycloalkenyl-substituted carbamates did not require a Rh(II) catalyst and proceeded via an iminoiodinane intermediate. The resulting tricyclic aziridines underwent ring opening when treated with various nucleophiles to give anti-derived products as expected for nucleophilic attack at the three-membered ring. The iodine(III)-mediated reaction of a 3-indolyl substituted carbamate, however, required a Rh(II) catalyst. The expected aziridine was not observed, but rather simultaneous spirocyclization of C(3) and stereoselective syn-acylation at C(2) occurred to give compound 41, whose structure was unequivocally established by an X-ray crystallographic study. The reaction proceeds in a stepwise manner via a metal-free zwitterionic intermediate which is attacked by a nucleophilic reagent on the same side of the amide anion. Related reactions occurred with both a 2-indolyl- and 3-benzofuranyl-substituted carbamate but with lower stereoselectivity. PMID- 15357599 TI - Study of the tautomeric equilibrium of pyridoxine in 1,4-dioxane/water mixtures by 13C nuclear magnetic resonance. Thermodynamic characterization and solvent effects. AB - A significant temperature dependence has been found for the (13)C NMR chemical shifts of pyridoxine in 10%, 20%, 30%, 40%, 50%, and 60% v/v 1,4-dioxane/water mixtures (pH = 7.0). The nuclei most sensitive to the temperature effect were C-3 and C-6 in all of the mixtures. This dependence has been explained on the basis of a thermally induced tautomeric equilibrium shift between the neutral and the dipolar forms of the pyridoxine molecule. The thermodynamic characterization of this tautomeric equilibrium, which interconverts quickly on the NMR time scale, has been achieved by considering the observed average (13)C NMR chemical shifts at different temperatures through fitting the experimental data to a theoretical curve. The fitting accuracy is greatly improved on using linear correlations between the average chemical shifts obtained from different nuclei at the same temperature. The methodology outlined above allows the DeltaH degrees value to be calculated for the tautomeric process and the chemical shifts of the pure extreme forms, i.e., neutral and dipolar, to be deduced. These values have been used to calculate the thermodynamic parameters of the tautomerization equilibrium in each dioxane/water mixture. The effect of solvent on the tautomeric equilibrium and the averaged chemical shift has been explained in terms of a multiparameter equation developed by Kamlet and Taft. The overall solvent effect is the sum of two different effects: the dipolarity and polarizability of the solvent and the ability of the solvent to act as a hydrogen-bond donor toward a solute. PMID- 15357600 TI - Multinuclear calixarene synthons with covalently linked aryl-palladium(II) complexes. AB - New synthetic procedures have been developed for potentially useful metallacalixarene building blocks. The metal sites were covalently connected to calix[n]arenes (n = 4, 6) by oxidative addition of 4-iodobenzyl precursors to either Pd(PPh(3))(4) or Pd(2)(dba)(3)/tmeda (dba = dibenzylideneacetone) to furnish calixarene-modified aryl-Pd(II)I(L(n)()) complexes [L(n)() = bis-PPh(3) or N,N,N',N'-tetramethylethylenediamine (tmeda)]. Methods were explored for the selective preparation of mono-Pd(II)-calix[4]arene and di-Pd(II)-calix[n]arenes (n = 4 or 6) complexes and also for bifunctional calix[4]arene synthons with two Pd(II) complexes accompanied by 4-pyridylmethyl or 4-cyanobenzyl groups. The properties of the Pd(II)-calix[n]arenes were studied in detail by one- and two dimensional NMR and mass spectrometric techniques. The X-ray molecular structures of two 4-iodobenzylcalix[4]arene precursors were also determined. PMID- 15357601 TI - Meso-substituted [34]octaphyrin(1.1.1.0.1.1.1.0) and corrole formation in reactions of a dipyrromethanedicarbinol with 2,2'-bipyrrole. AB - The reaction of a dipyrromethanedicarbinol with 2,2'-bipyrrole leading to meso substituted [34]octaphyrin(1.1.1.0.1.1.1.0) and/or corrole was investigated to determine the effect of key reaction parameters on the distribution of the two macrocycles. Solvent, acid catalyst, acid quantity, oxidant, oxidant quantity, and reaction time were surveyed for a model reaction affording 5,10,19,24,29,38 hexaphenyl[34]octaphyrin(1.1.1.0.1.1.1.0) (HPO) and/or meso-triphenylcorrole (TPC). HPO was found to be a fairly ubiquitous product, produced in yields as high as 23% (UV-vis), while TPC was observed infrequently, in yields up to 10% (UV-vis). A preparative-scale reaction provided HPO in an isolated yield of 25%. The methodology was extended to the synthesis of an octaphyrin bearing two different substituents in defined locations and to an octaphyrin possessing electron-withdrawing pentafluorophenyl substituents. Preferential formation of octaphyrin instead of corrole suggests that the anti conformation of 2,2' bipyrrole is the relevant form under the reaction conditions surveyed. The spectral properties of the novel meso-substituted [34]octaphyrin(1.1.1.0.1.1.1.0) species are similar to those of the known beta-substituted analogue, including spectra consistent with the absence of macrocycle aromaticity despite a main conjugation path of 34 pi-electrons. Key to the overall study was the development of a refined synthesis of 2,2'-bipyrrole. PMID- 15357602 TI - Sulfur dioxide mediated one-pot, three- and four-component syntheses of polyfunctional sulfonamides and sulfonic esters: study of the stereoselectivity of the ene reaction of sulfur dioxide. AB - The ene reaction of sulfur dioxide with enoxysilanes or with allylsilanes generates silyl sulfinates that can be brominated (Br(2) or NBS) or chlorinated (NCS or Cl(2)) to produce the corresponding sulfonyl halides. They react with primary and secondary amines or alcohols to give the corresponding sulfonamides and sulfonic esters, respectively. The hetero-Diels-Alder addition of sulfur dioxide to 1-oxy- or 1,3-dioxy-1,3-dienes generates zwitterions that add to enoxysilanes or allylsilanes giving silyl sulfinates that can be converted in situ into polyfunctional sulfonamides or sulfonic esters. This realizes quick access to libraries of complicated sulfonamides and sulfonic esters applying one pot, three- and four-component methods. PMID- 15357603 TI - Novel Kumada coupling reaction to access cyclic (2-azaallyl)stannanes. Cycloadditions of cyclic nonstabilized 2-azaallyllithium species derived from cyclic (2-azaallyl)stannanes. AB - A Kumada cross-coupling reaction involving organomagnesium reagents and (3 methylthio-2-azaallyl)stannanes with a Ni(0) catalyst provided cyclic nonstabilized (2-azaallyl)stannanes in moderate to good yields. Primary alkyl, aryl, and allylic organomagnesium reagents can be used as the cross-coupling partner. In general, NiCl(2)dppp in toluene at room temperature provided the shortest reaction times and most consistent yields. The azomethine ylides and 2 azaallyllithium species derived from these stannanes were shown to undergo efficient [3 + 2] cycloaddition reactions to provide azabicyclo[n.2.1]alkanes as the endo cycloadducts. These cycloadducts were found to be useful as starting materials for further elaboration into aza-bridged bicyclic natural and unnatural products of biological interest. Although cyclic 2-azaallyllithium species have been generated previously, this work reports the first generation and cycloaddition of entirely nonstabilized 2-azaallyllithium species. In addition a novel extension of the Kumada coupling was developed to allow for the preparation of the cyclic (2-azaallyl)stannanes, which are precursors to the nonstabilized 2 azaallyllithium species. PMID- 15357604 TI - Sequential reactions of trimethylstannyl anions with vinyl chlorides and dichlorides by the SRN1 mechanism followed by palladium-catalyzed cross-coupling processes. AB - The reactions of trimethylstannyl ions (Me(3)Sn(-)) with vinyl chlorides in liquid ammonia give good yields of vinylstannanes. Some of them react in the dark, and others need light stimulation to react. The fact that these reactions are inhibited by radical and radical anion traps shows that they occur by the S(RN)1 mechanism. When the reaction takes place with 1,1-dichloro-1-alkenes, monosubstitution reduced products are formed in an E/Z mixture. The efficient synthesis of triarylolefins by Pd(0)-catalyzed cross-coupling reactions of vinylstannanes with several iodoarenes is reported. Similar yields were obtained in one-pot-type reactions. PMID- 15357605 TI - First synthesis and structural elucidation of (-)-presphaerene. AB - The first total synthesis of (-)-presphaerene (1) was achieved from (R) glyceraldehyde 9 in 19 steps, demonstrating the novel "folding and allylic strain controlled" intramolecular ester enolate S(N)2' alkylation strategy could be extended to the stereoselective synthesis of cyclopentanoid natural products. The present study also established the relative and absolute stereochemistry of 1, and the absolute structures of co-occurring sphaeroanes from the red alga Sphaerococcus coronopifolius. PMID- 15357606 TI - Synthetic studies aimed at (-)-cochleamycin A. Evaluation of late-stage macrocyclization alternatives. AB - An efficient route to the fully functionalized ABC ring systems of the unnatural enantiomer of cochleamycin A was developed. L-(-)-Malic and L-(-)-ascorbic acids served well as starting materials for the two building blocks used to construct an (E,Z,E)-1,6,8-nonatriene intermediate. The AB part structure was assembled by way of a stereocontrolled intramolecular Diels-Alder cycloaddition via adoption of an endo transition state. From this vantage point, two general pathways were subsequently explored as to their suitability for elaboration of the CD rings. Initially examined was a protocol involving 10-membered carbocycle construction. When this approach was demonstrated not to be workable, attention was directed to 10-membered macrolactonization as an alternative tactic. Although assembly of the C-ring in this manner was easily achieved, ultimate closure of the six-membered ring to form ring D remains an unsolved problem. PMID- 15357607 TI - Development of N-benzamidothioureas as a new generation of thiourea-based receptors for anion recognition and sensing. AB - A series of neutral N-(substituted-benzamido)-N'-phenylthioureas (substituent = p OC(2)H(5), p-CH(3), m-CH(3), H, p-Cl, p-Br, m-Cl, and p-NO(2)) were designed as anion receptors, in which the thiourea binding site was attached to the benzamido moiety via an N-N bond. The absorption spectra of these N-benzamidothioureas in acetonitrile peaked at ca. 270 nm were found to show unprecedented red shifts by 7 373 to 14 325 cm(-1) in the presence of anions such as AcO(-), F(-), and H(2)PO(4)(-). Under the same conditions, the classic neutral thiourea receptors, N-(substituted-phenyl)-N'-phenylthioureas, showed absorption spectral shifts in most cases of less than 800 cm(-1) with one exception of 6501 cm(-1). Control experiments, effects of protic solvent, and (1)H NMR titration confirmed the formation of hydrogen-bonding complexes between the new N-benzamidothiourea receptors and anions. The binding constants with AcO(-), for example, are at 10(5)-10(7) mol(-1) L order of magnitude, which are 13 to 590 times those of the corresponding classic N-phenylthioureas in the same solvent. It was found that, whereas the absorption of the N-benzamidothiourea receptors showed essentially no dependence on the substituent, the substantially red-shifted new absorption band of the N-benzamidothiourea-anion binding complex was sensitively subject to the substituent. A linear relationship was found between the absorption energies of the N-benzamidothiourea-acetate binding complexes and the Hammett constants of the substituents with a negative slope of -0.34 eV. This led to the assignment that the substantially red-shifted absorption band was the ground-state intramolecular charge-transfer absorption with the substituent locating in the electron acceptor moiety. It was concluded that anion binding to the thiourea moiety of the N-benzamidothiourea receptors switched on their ground-state charge transfer. An anion-binding induced structural change was suggested to occur around the N-N bond in N-benzamidothioureas, which resulted in a substantially increased electron donating ability of the electron donor in the receptor molecules. As a consequence, the ground-state charge transfer takes place in the N-benzamidothiourea-anion binding complexes, leading to unprecedented red shifts in the absorption spectra and substantially enhanced anion binding affinities than those of the corresponding N-phenylthiourea receptors. N-Benzamido-N' phenylthioureas represent a new generation of neutral thiourea-based anion receptors that show substantially improved anion binding performance important for anion sensing and recognition. PMID- 15357608 TI - Derivative of alpha,beta-dicyanostilbene: convenient precursor for the synthesis of diphenylmaleimide compounds, E-Z isomerization, crystal structure, and solid state fluorescence. AB - A convenient and efficient procedure was developed for preparing 3,4-diaryl substituted maleimides through the improved synthesized diaryl-substituted fumaronitrile. The synthesis of diphenyl-substituted fumaronitrile derivatives from phenylacetonitrile compounds was analyzed and improved. We found the stoichiometry of the sodium methoxide and the concentration of the starting material, phenylacetonitrile derivatives, were crucial for the high yield and easy purification of the products. Particularly, bis(4-bromophenyl)fumaronitrile, bis(3-trifluoromethylphenyl)fumaronitrile, and bis(4-methoxyphenyl)fumaronitrile were isolated in good yields of 70-90% by simple suction filtration. In addition, (1)H NMR provided compelling evidence that the E-Z isomerization was involved in the formation reaction of the maleimide compounds from either fumaronitrile or maleonitrile derivatives. Single-crystal X-ray structures of these three fumaronitrile derivatives, the first three of the kind, were obtained, revealing the nonplanar molecular structure. We ascribe the strong solid-state fluorescence of these diphenylfumaronitrile derivatives to the nonplanar structure that inhibits the close packing of the molecule aggregation and thus the fluorescence quenching. PMID- 15357609 TI - Highly regio- and stereoselective synthesis of polysubstituted cyclopropane compounds via the Pd(0)-catalyzed coupling-cyclization reaction of 2-(2',3' allenyl)malonates with organic halides. AB - A new method for highly regio- and stereoselective synthesis of polysubstituted cyclopropane compounds via the Pd(0)-catalyzed coupling-cyclization reaction of 2 (2',3'-allenyl)malonates with organic halides is described. In these reactions, the starting materials are easily available and the operation is convenient. The ratios of trans-isomer/cis-vinylic cyclopropanes are up to 98:2. PMID- 15357610 TI - First diastereoselective intramolecular Baylis-Hillman reaction: an easy access to chiral alpha-methylene-beta-hydroxylactones. AB - The first diastereoselective intramolecular Baylis-Hillman reaction of chiral substrates is reported wherein both aldehyde and activated olefin coexist as substituents to afford alpha-methylene-beta-hydroxylactones in good yields exclusively as single isomers under the standard base-catalyzed reaction conditions in CH(2)Cl(2). Formation of alkoxylactones by an in situ derivatization of adducts was also observed. PMID- 15357611 TI - Synthesis of tetrasubstituted ozonides by the Griesbaum coozonolysis reaction: diastereoselectivity and functional group transformations by post-ozonolysis reactions. AB - The diastereoselectivity of the Griesbaum coozonolysis reaction with O-methyl 2 adamantanone oxime and 4-substituted cyclohexanones reveals that the major tetrasubstituted ozonide isomers possess cis configurations, suggesting a preferred axial attack of the carbonyl oxide on the cyclohexanone dipolarophiles. It is evident that these tetrasubstituted ozonides are quite stable to triphenylphosphine, borohydrides, hydrazine, alkyllithiums, Grignard reagents, mercaptides, and aqueous KOH as illustrated by the synthesis of amine, alcohol, acid, ester, ether, sulfide, sulfone, and heterocycle-functionalized ozonides by a wide range of post-ozonolysis transformations. PMID- 15357612 TI - A facile synthesis of C2,N3-disubstituted-4-quinazolone. AB - A simple and efficient methodology for the synthesis of C(2),N(3)-disubstituted-4 quinazolones from anilines and N-acylanthranilic acids was developed. The new cyclization conditions are much milder than any other reported protocols and resulted in excellent yields (87-98%) without chromatography. PMID- 15357613 TI - Formation of a quaternary carbon center through the Pd(0)/PhCOOH-catalyzed allylation of cyclic beta-keto esters and 1,3-diketones with alkynes. AB - Formation of a quaternary carbon center through the allylation of beta-keto esters and 1,3-diketones with alkynes is accomplished by the use of Pd(0)/benzoic acid catalyst. Reactions of various cyclic beta-keto esters and 1,3-diketones with alkynes in the presence of Pd(2)dba(3).CHCl(3) (5 mol %), PPh(3) (40 mol %), and PhCOOH (10 mol %) proceeded at 100 degrees C in toluene (5 M) to give the corresponding allylation products in high yields in a regio- and stereoselective manner. The possibility of asymmetric allylation is also discussed. PMID- 15357614 TI - Universal reusable polymer support for oligonucleotide synthesis. AB - A new efficient reusable universal polymer support for oligonucleotide synthesis, based on a non-ammoniacal cleavable linker, is described. Twenty six cycles of oligonucleotide synthesis have been carried out without compromising the quality of the fully deprotected oligonucleotides. PMID- 15357615 TI - Unexpected formation of tetrasubstituted 2,3-dihydrofurans from the reactions of beta-keto polyfluoroalkanesulfones with aldehydes. AB - Catalyzed by piperidine, the reactions of beta-keto polyfluoroalkanesulfones with aromatic aldehydes afforded the unexpected tetrasubstituted 2,3-dihydrofurans in good yields, probably proceeding through the normal Knoevenagel condensation products. This reaction provided an efficient and novel method for the stereoselective synthesis of fluorine-containing tetrasubstituted trans-2,3 dihydrofurans. PMID- 15357616 TI - Asymmetric synthesis of beta-hydroxy amino acids via aldol reactions catalyzed by chiral ammonium salts. AB - The Cinchona alkaloid derived chiral ammonium salt developed by Park and Jew functions as an effective catalyst for the synthesis of beta-hydroxy alpha-amino acids via asymmetric aldol reactions under homogeneous conditions. The syn diastereomers are obtained in good ee, and aryl-substituted aliphatic aldehydes are the best substrates for the reaction. These results represent the highest ee's obtained to date in direct aldol reactions of glycine equivalents catalyzed by inexpensive, readily prepared chiral ammonium salts. PMID- 15357617 TI - A chiral synthesis of (-)-spiro[1-azabicyclo[2.2.2]octane-3,5'- oxazolidin-2' one]: a conformationally restricted analogue of acetylcholine that is a potent and selective alpha7 nicotinic receptor agonist. AB - A direct, short chiral synthesis of the selective alpha7 nicotinic receptor agonist (-)-spiro[1-azabicyclo[2.2.2]octane-3,5'-oxazolidin-2'-one] (AR-R17779) is presented. The key step utilized attack of the dianion of the (R)-HYTRA ester [(R)-(+)-2-hydroxy-1,2,2-triphenylethyl acetate] on quinuclidin-3-one, followed by a selective precipitation of the diasteriomeric tertiary alcohol that led to (S)-(-)-AR-R17779 in two additional steps. PMID- 15357618 TI - Acrylamide in the Baylis-Hillman reaction: expanded reaction scope and the unexpected superiority of DABCO over more basic tertiary amine catalysts. AB - DMAP, DBU, and quinuclidine efficiently promote novel hydroalkoxylation reactions of acrylamide in primary alcohol solvents. DABCO is a comparatively poor hydroalkoxylation promoter and can effect clean, selective Baylis-Hillman reactions between acrylamide and aldehydes in alcoholic/aqueous media in which more basic nucleophilic catalysts promote hydroalkoxylation preferentially. Optimization of the reaction conditions has allowed acrylamide to be reacted with a range of aromatic aldehydes in moderate to excellent yields, including the first examples involving deactivated, electron-rich substrates such as p tolualdehyde and o-anisaldehyde. PMID- 15357619 TI - Preparation of asymmetric alpha-ketophosphonates by [3,3]-sigmatropic shift of enolphosphonates. AB - Alpha-ketophosphonates are prepared by a [3,3]-sigmatropic shift of enolphosphonates. PMID- 15357620 TI - Solvent free amination reactions of aryl bromides at room temperature catalyzed by a (pi-allyl)palladium complex bearing a diphosphinidenecyclobutene ligand. AB - An air- and moisture-stable (pi-allyl)palladium complex bearing a unique diphosphinidenecyclobutene ligand effectively catalyzes amination reactions of aryl bromides with amines, where the reactions proceed under mild conditions without solvent, with 2 mol % of catalyst and 1 equiv of t-BuOK at room temperature. Under these conditions the amination products were obtained in good to excellent isolated yields. PMID- 15357621 TI - Intramolecular nitrone cycloaddition reaction on carbohydrate-based precursors: application in the synthesis of spironucleosides and spirobisnucleosides. AB - A simple synthesis of chiral spironucleosides and spirobisnucleosides is described. Intramolecular 1,3-dipolar nitrone cycloaddition reaction of d-glucose derived precursors having olefin at C-3 and nitrone at C-5, C-1, or C-2 (in nor series) furnished bisisoxazolidinospirocycles 4-7, 11, and 12 in good yields. Reductive ring opening of the isoxazolidine moieties in 4-6 followed by construction of a nucleoside base upon the generated amino groups smoothly yielded spirobisnucleosides 17 and 18 and spironucleosides 20 and 21. PMID- 15357622 TI - Syntheses of very dense halogenated liquids. AB - A family of halogenated liquids with densities ranging from 1.95 to 2.80 g cm(-3) was readily synthesized by a one-pot procedure. These liquids exhibit characteristics of ionic liquids with melting/transition points lower than room temperature, long liquid ranges, and marked hydrolytic and thermal stabilities. PMID- 15357624 TI - Correlations between factors determining the pharmacokinetics and antiviral activity of HIV-1 non-nucleoside reverse transcriptase inhibitors of the diaryltriazine and diarylpyrimidine classes of compounds. AB - OBJECTIVE: To investigate the important factors that determine the bioavailability and the antiviral activity of the diaryltriazine (DATA) and diarylpyrimidine (DAPY) non-nucleoside reverse transcriptase inhibitors (NNRTIs) of HIV-1 in animal species and humans using cell-based assays, physicochemical and computed parameters. METHODS: This naturalistic study included 15 parameters ranging from molecular mechanics calculations to phase I clinical trials. The calculated parameters were solvent-accessible surface area (SASA), polar surface area and Gibbs free energy of solvation. Physicochemical parameters comprised lipophilicity (octanol/water partition coefficient [cLogP]), ionisation constant (pKa), solubility and aggregate radius. Cell-based assays included human colonic adenocarcinoma cell (Caco-2) permeability (transepithelial transport), drug metabolism and antiviral activity (negative logarithm of the molar effective concentration inhibiting viral replication by 50% [pEC50]). Exposure was tested in rats, dogs and human volunteers. RESULTS: Of the 15 parameters, eight correlated consistently among one another. Exposure (area under the plasma concentration-time curve [AUC]) in humans correlated positively with that in rats (r = 1.00), with transepithelial transport (r = 0.83), lipophilicity (r = 0.60), ionisability (r = 0.89), hydrodynamic radius of aggregates (r = 0.66) and with antiviral activity (r = 0.61). Exposure in humans was also seen to correlate negatively with SASA (r = -0.89). No consistent correlation was found between exposure in dogs and the eight parameters. Of the 14 DATA/DAPY molecules, 11 form aggregates with radii between 34 and 100 nm. CONCLUSIONS: We observed correlations between exposure in humans with exposure in rats, transepithelial transport (Caco-2 cells), ionisability, lipophilicity, aggregate radius and SASA in the class of DATA/DAPY NNRTI compounds. The lipophilic DATA/DAPY compounds form aggregates. It can be assumed that absorption in the intestinal tract and endocytosis in infected cells of these lipophilic compounds are governed by the common phenomenon of aggregate formation. As the lymphatic system offers a pathway for intestinal uptake of aggregates, this may offer a therapeutic advantage in the treatment of HIV-1 infection. Although it was not the objective of the study, we found that the rat was a better in vivo model than the dog for the prediction of systemic exposure in this particular set of compounds. PMID- 15357625 TI - Oral pyridoxine during pregnancy : potential protective effect for cardiovascular malformations. AB - OBJECTIVE: To study the human risk and benefit of oral pyridoxine (vitamin B6) treatment during pregnancy. DESIGN AND SETTING: The analysis of cases with 25 congenital abnormality (CA) groups and their all-matched controls without CAs in the population-based dataset of the large Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1996. STUDY PARTICIPANTS: 22,843 cases of pregnant women who had newborns or fetuses with CAs and 38,151 pregnant women who had newborn infants without any CAs (control group). MAIN OUTCOME MEASURES: Prevalence of pyridoxine use in early pregnancy among mothers of cases with different CAs and control mothers with infants without any CA. RESULTS: 2013 (8.8%) case mothers and 4086 (10.7%) control mothers were treated with pyridoxine (adjusted prevalence odds ratio [POR] 0.8; 95% confidence interval [CI] 0.7, 0.9). The analysis of cases with different defects and their all-matched controls did not indicate any obvious teratogenic potential of pyridoxine use during the second and third months of gestation, i.e. in the critical period for the development of most major CAs. However, some protective effect was found for cardiovascular malformations (adjusted POR 0.8; 95% CI 0.7, 0.9). CONCLUSION: Treatment with pyridoxine during pregnancy does not indicate a teratogenic risk to the fetus, but may provide some protective effect for cardiovascular malformations. PMID- 15357626 TI - Bioequivalence of two recombinant interferon alpha-2b liquid formulations in healthy male volunteers. AB - OBJECTIVE: Interferon (IFN) alpha-2b is a protein with antiviral, antiproliferative and immunoregulatory properties that is approved for several clinical indications. A new liquid, albumin-free, IFNalpha-2b formulation has recently been developed. This study aimed to evaluate the equivalence of the pharmacokinetic, pharmacodynamic and safety properties of the new formulation with a reference one in healthy male volunteers. METHODS: A randomised, crossover, double-blind study with a 3-week washout period was performed in which Heberon Alfa R (formulation A) and Viraferon (formulation B) were compared. A single 20 x 10(6) IU IFNalpha-2b dose was administered subcutaneously to 14 apparently healthy male subjects. Serum IFN level was measured over 48 hours by enzyme immunoassay (EIA) and by antiviral activity titration. Clinical and laboratory variables were determined, as were pharmacodynamic and safety criteria. RESULTS: Groups were homogeneous with regard to all demographic and baseline variables. Pharmacokinetic comparison by EIA did not show differences between the formulations: area under the curve (AUC) 2572 versus 2561 ng x h/L, maximum plasma concentration (Cmax) 318 versus 354 ng/L, time to Cmax (tmax) 8.2 versus 8.5 h, elimination half-life (t(1/2)) 5.87 versus 6.08 h, terminal elimination rate (lambda) 0.122 versus 0.118 h(-1), and mean residence time (MRT) 10.9 versus 12.0 h for formulations A and B, respectively. The differences never reached 20%, which is the clinically significant threshold. The 90% confidence interval of the ratio between them was in all cases within the 0.8, 1.25 range. The two formulations were clinically equivalent with regard to serum IFN antiviral activity titration (0.8, 1.25 criterion) regarding their pharmacokinetic parameters. There were no significant differences with respect to the pharmacodynamic variables: serum beta2-microglobulin and temperature increase. Heart rate and blood pressure changes did not differ either. Both products provoked similar haematological count decreases and had similar safety profiles. The most frequent adverse reactions were fever, tachycardia, headache and arthralgias. CONCLUSION: The overall analysis strongly suggests the bioequivalence of these two products. PMID- 15357627 TI - Effect of D-003, a mixture of very high molecular weight aliphatic acids, on prednisolone-induced osteoporosis in Sprague-Dawley rats. AB - BACKGROUND: Drugs inhibiting cholesterol biosynthesis may affect bone metabolism through inhibition of the mevalonate pathway resulting in the inhibition of protein prenylation required for osteoclast activity. D-003 is a mixture of high molecular weight aliphatic primary acids purified from sugar-cane (Saccharum officinarum) wax, with cholesterol-lowering effects demonstrated in experimental and clinical studies. D-003 inhibits cholesterol biosynthesis through indirect regulation of HMG-CoA reductase activity. A previous study demonstrated that D 003 prevented bone loss and bone resorption on ovariectomy-induced osteoporosis in rats. Corticosteroid-induced osteoporosis is the result of changes affecting calcium homeostasis, but the hallmark of corticosteroid-induced bone loss is the direct effects on bone cells, such as inhibition of osteoblastogenesis, promotion of apoptosis of osteoblasts and osteocytes, and decrease in bone formation. OBJECTIVE: To determine whether D-003 could prevent the bone loss induced with prednisolone in Sprague-Dawley rats. METHODS: Rats were randomly distributed in five groups (ten rats per group): a sham-operated control and four groups orally treated with prednisolone 6 mg/kg for 80 days; a positive control orally treated with vehicle; and three groups orally treated with D-003 at 5, 25 and 200 mg/kg, respectively. Rats were killed, bones removed and histological variables of bone resorption and formation studied for histomorphometry. RESULTS: Compared with the sham group, prednisolone significantly (p < 0.01) reduced trabecular bone volume (TBV), while D-003 significantly (p < 0.001) and dose-dependently prevented the prednisolone-induced reduction of TBV. Treatment with prednisolone lowered (p < 0.001) trabecular thickness (TbTh) and number (TbN), while increasing (p < 0.001) the gap between trabeculae. D-003 (5, 25 and 200 mg/kg/day) significantly (p < 0.001) and dose-dependently prevented the reduction of TbTh and TbN and the increase of trabecular gap induced with prednisolone. Treatment with prednisolone increased both the surface and number of osteoclasts compared with sham (p < 0.001). D-003 (5-200 mg/day), however, prevented this effect (p < 0.001 for all comparisons). D-003 also prevented (p < 0.001) the reduction of osteoblast surface (ObS/BS) induced by prednisolone. Osteonecrotic areas were observed in all positive controls, but in none of the sham animals. Positive controls showed hypertrophy of bone marrow adipocytes and lipid-laden pluripotential stromal cells in bones. A significant and dose-dependent reduction of the frequency of animals showing prednisolone-induced osteo-necrosis was observed across the doses of D-003 (5, 25 and 200 mg/kg) investigated here. CONCLUSIONS: D-003 (5, 25 and 200 mg/kg) prevented trabecular bone loss and femoral neck osteonecrosis induced with prednisolone in Sprague Dawley rats, also increasing osteoblast surface and reducing bone resorption parameters. These results suggest that D-003 could be useful for managing corticosteroid-induced osteoporosis. PMID- 15357628 TI - Bleomycin--electrical pulse delivery: electroporation therapy-bleomycin- Genetronics; MedPulser-bleomycin--Genetronics. AB - Genetronics Biomedical is using its electroporation therapy technology to deliver bleomycin to tumour cells for the treatment of cancer. Genetronics have developed the MedPulser Electroporation Therapy System, which consists of an electrical pulse generator and disposable electrode applicators. The MedPulser system enables the delivery of large molecules into cells by briefly applying an electric field to the cell. This causes a transient permeability in the cell's outer membrane characterised by the appearance of pores across the membrane. After the field is discontinued, the pores close, trapping the therapeutic molecules inside the target cells. Genetronics is using the MedPulser System in conjunction with bleomycin, an antineoplastic antibiotic that binds to DNA causing strand scissions. Genetronics is seeking a licensing partner for the use of electroporation for the delivery of drugs in chemotherapy. In 1998, Genetronics entered a licensing and development agreement with Ethicon for electroporation and electrofusion. Under the terms of this agreement, Ethicon was to develop and clinically test the Genetronics electroporation delivery system and conduct all regulatory activities throughout the world except Canada. Ethicon would also market the products once regulatory approval has been obtained and Genetronics was to receive a percentage of the net sales and as license fees. However, in July 2000, Ethicon exercised its rights to terminate the agreement without cause. All rights were returned to Genetronics in January 2001. In 1997, Genetronics entered an agreement with Abbott Laboratories for the manufacture of bleomycin for use in the US in its MedPulsar system after regulatory approval had been granted for its use in the treatment of solid tumours. In a separate supply agreement, Faulding Inc. has agreed to manufacture bleomycin for Genetronic for use in Canada after regulatory approval had been granted. The MedPulsar Electroporation Therapy System with bleomycin is currently in phase III pivotal studies in the US as a treatment for recurrent and second primary squamous cell carcinomas of the head and neck. Genetronics received approval for the Electroporation Therapy system as a device in March 1999 when it achieved CE Mark certification. In February 2004, Genetronics announced that it had completed a Special Protocol Assessment review process with the US FDA for two new trials that will compare bleomycin electroporation therapy to surgery. The primary endpoint will be tissue and function preservation rather than survival. One proposal is for recurrent head and neck cancer, and the other is for disfiguring cutaneous cancer. Three Institutional Review Boards in the US have approved the two protocols and Genetronics has initiated enrollment. In June 2004, Genetronics was granted fast-track status for its MedPulsar Electroporation Therapy System clinical development programme for patients with head and neck cancer. Shifting from a primary endpoint of survival to a quality-of-life outcome will enable those clinical trials to be carried out faster with less cost and with a higher likelihood of success. As a result, Genetronic's phase III trials focussing on survival as a primary endpoint have been discontinued. This includes a phase III trial for late-stage, recurrent head and neck cancer in combination with the normal standard of treatment compared with normal standard of treatment alone. Interim results from this trial had suggested bleomycin electroporation therapy demonstrated local tumour control and preservation of organ function, as well as non-inferiority when compared with surgery. This trial was initiated in May 2002. In March 2004, Genetronics initiated a post-European regulatory approval clinical study in patients with primary or recurrent squamous cell carcinoma of the head and neck (SCCHN). This study aims to enroll approximately 100 patients at 12-15 hospitals located in the UK, Germany, Italy, France, Austria and other western European countries. The study is designed to support the commercialisation of the MedPulser Electroporation System in the EU. Prior clinical trials established the safety and performance of the MedPulser System for the treatment of SCCHN, leading to approval for sale in the EU based on achieving the CE Mark. This study will document the clinical and pharmacoeconomic benefit in support of reimbursement approval throughout Western Europe, establish centres of excellence to facilitate early sales, create a reference and customer base for a projected European commercial launch in 2005, and generate safety and efficacy data to support marketing applications in the US. The bleomycin delivery system has completed phase IIB trials in the US, Canada and Europe in patients with squamous cell carcinoma of the head and neck who have failed conventional therapies. Phase II data were submitted to the FDA in the first quarter of 2002 and a phase III trial was launched in May 2002. The therapy is also being used in France in patients with cancers of the head and neck, liver (metastatic) and melanoma. A review of the data from these phase II trials was completed in April 2001. In June 2004, Genetronics was granted two US patents. US patent 6,748,265 covers its trans-surface drug and gene delivery technology and provides additional proprietary rights for an apparatus and method to deliver genes, drugs and other molecules through tissue surfaces. The second US patent, 6,746,441, pertains to the field of ex vivo therapies and covers the introduction of molecules into cells by electroporation, either in a continuous-flow or batch mode, with a variable electric field orientation. In July 2004, Genetronics received a US patent (no. 6,763,264) covering methods for the in vivo delivery of a recombinant expression vector (DNA) or a pharmaceutical agent into tissue cells, and a method for the therapeutic application of electroporation to a patient to introduce macromolecules. PMID- 15357629 TI - Mismatched double-stranded RNA: polyI:polyC12U. AB - Ampligen [polyI:polyC12U] is a mismatched double-stranded RNA that acts by inducing interferon production (immunomodulator) and by activating an intracellular enzyme (RNase-L) against viral RNA transcripts (antiviral). Ampligen, currently under development by Hemispherx Biopharma in the US, acts on the immunological system through T-lymphocyte stimulation and is indicated for the treatment of chronic fatigue syndrome and acquired immunodeficiency deficiency syndrome (AIDS), as part of the combined therapy. Ampligen is available for licensing worldwide. In February 2004, Fujisawa Deutschland GmbH, a subsidiary of Fujisawa Pharmaceutical Co., entered into an option agreement with Hemispherx Biopharma with the intent of becoming a distributor for Ampligen for the potential treatment of chronic fatigue syndrome in Germany, Switzerland and Austria. An option fee of 400,000 euros was paid pursuant to the terms of the option agreement and upon execution of the Distribution Agreement, Fujisawa will pay Hemispherx fees and milestone payments with a potential worth of several millions of dollars. In September 2003, Hemispherx Biopharma Inc. entered into an agreement with Guangdong Medicine Group Corporation to organise clinical trials, marketing, sales and distribution for both of its lead compounds, Ampligen and Alferon N in the People's Republic of China. The agreement stipulates that the Guangdong Medicine Group Corporation (GMC) will conduct clinical trials with Ampligen for the treatment of HIV. All costs related to the trials are to be covered by GMC. Additionally, GMC has to develop and implement marketing and promotional programmes. In May 2003, Hemispherx Biopharma and the Center for Cell and Gene Therapy entered into a research project agreement that will see Ampligen implemented in a protocol used in patients with relapsed EBV-positive Hodgkin's Lymphoma. In March 2002, Esteve and Hemispherx Biopharma entered into a collaborative agreement under which Esteve will be the sole distributor of Ampligen in Spain, Portugal and Andorra for the treatment of chronic fatigue syndrome. Under this agreement, in addition to other terms, Esteve will also collaborate in the drug product development by conducting clinical studies in Spain in patients coinfected with HIV/HCV. In July 2001 Hemispherx Biopharma announced that it had formed a strategic alliance with Empire Health Resources for clinical trials of Ampligen in the treatment of HIV and hepatitis C virus infections. Empire Health Resources, a healthcare management firm, will be responsible for accrual and retention of patients for HIV trials, and protocols for trials in patients with hepatitis C or both HIV and hepatitis C infections. Hemispherx has entered into a collaboration with RED Laboratories, and RED Laboratories NV expects that this will facilitate the continued development of Ampligen. Hemispherx has also entered into an agreement with Schering Plough to use a Schering facility as its principal manufacturing platform in the US. This agreement may be expanded to include other territories. Hemispherx and AOP Orphan Pharmaceuticals have signed a marketing agreement for Ampligen for the treatment of chronic fatigue syndrome for Austria, the Czech Republic, Poland and Hungary. In an arrangement between Hemispherx and Bioclones, Bioclones has certain marketing rights for Ampligen in the Southern Hemisphere, UK and Ireland. In the US, Ampligen has been granted orphan drug status for the treatment of AIDS, renal cell carcinoma (phase II, completed), chronic fatigue syndrome (phase III) and invasive/metastatic malignant melanoma (phase II). In August 2004, Hemispherx announced that it intends to use the proceeds from the private placement of company stock to complete the clinical work for its immunotherapeutics/ antivirals Ampligen and Oragens. Previously, Hemispherx submitted an application to the EMEA for the approval of Ampligen for the treatment of chronic fatigue syndrome; the first stage of th;) for the treatment of chronic fatigue syndrome; the first stage of the regulatory review has been cleared. In 2000, Hemispherx Europe (Hemispherx) obtained orphan drug status for Ampligen for the treatment of chronic fatigue syndrome in the EU, providing Hemispherx with 10 years of marketing exclusivity following the launch of the drug, as well as potential financial research benefits for the agent. In February 2000, Crystaal Corporation (now Biovail Pharmaceuticals Canada) acquired exclusive marketing rights to Ampligen in Canada, where it submitted an NDA for the agent for the treatment of chronic fatigue syndrome. In the meantime, Ampligen has been available since May 1996 under the Canadian Emergency Drug Release Programme for the treatment of chronic fatigue syndrome and immune dysfunction syndrome by Rivex Pharma (Helix BioPharma). Bioclones has initiated clinical studies with Ampligen for the treatment of chronic fatigue syndrome in Australia. The active substance for Ampligen is manufactured by F.H. Faulding Ltd. Clinical treatment programmes for chronic fatigue syndrome in other Pacific Rim countries are planned. Ampligen is available for severe chronic fatigue syndrome on a named patient, cost-recovery basis in South Africa. Hemispherx has developed a 'ready-to-use' liquid formulation of the drug and has begun treating patients with chronic fatigue syndrome in ongoing clinical trials. Hemispherx has also developed an oral version of the drug (Oragen), which is undergoing preclinical evaluation. In February 2001, Hemispherx Biopharma announced that it was initiating phase II/III trials of Ampligen in the treatment of late-stage, multidrug-resistant strains of HIV in the European Union. Patients treated in these studies will have exhausted all other treatment options. In July 2001, Hemispherx stated that Ampligen was being evaluated in a phase IIb trial in patients with HIV in the US. The trial, comprising two studies, REARMI and REARMII (Research/Evaluation of Ampligen for Retroviral Mutations I and II), will evaluate the ability of Ampligen to prevent the emergence of mutated, drug-resistant strains of the virus. 'Several hundred' patients currently on antiretroviral therapy and at risk of viral relapse will be enrolled at centres in Connecticut, New York, Florida and California. A second phase IIb study evaluating the effect of Ampligen on structured treatment interruptions (STI) is also underway. Final results from this study were reported in December 2002. NIH sponsored studies of potential therapies for SARS have identified Ampligen as having unusually high and consistent antiviral activity against human coronavirus, the pathogen implicated as the causative agent of the disease. Ampligen demonstrated very high potency at very low concentrations (0.4 microg/mL) and had a favourable safety profile. In October 2003, Hemispherx announced that, based on these promising new results, the company will stockpile injectible and/or oral formats of Ampligen and Alferon N. Independent researchers have demonstrated the antiviral activity of Ampligen against flaviviruses (West Nile virus, Equine Encephalitis virus, Dengue fever virus and Japanese Encephalitis virus) as well as virus classes associated with bioterrorism. In an animal study, Ampligen was shown to prevent destruction of nerve cells, reduce virus concentrations in the brain and blood stream and increase survival rates. Researchers at the Rega Institute in Belgium have published results from an animal study demonstrating that Ampligen was superior at protecting mice against coxsackie B3 virus-induced myocarditis compared with pegylated interferon. In May 2004 Hemispherx announced that it had filed an expanded US patent application covering the use of Ampligen for the potential treatment and prevention of severe acute respiratory syndrome (SARS) and dreaded emerging viruses. PMID- 15357630 TI - Rubitecan: 9-NC, 9-Nitro-20(S)-camptothecin, 9-nitro-camptothecin, 9 nitrocamptothecin, RFS 2000, RFS2000. AB - Rubitecan [Orathecin, 9-nitrocamptothecin, 9NC, RFS 2000] is a topoisomerase I inhibitor extracted from the bark and leaves of the Camptotheca acuminata tree, which is native to China. Rubitecan is an oral compound being developed for the treatment of pancreatic cancer and other solid tumours by SuperGen. One of the major benefits of rubitecan is that it can be administered in an outpatient setting, so patients can be treated in their homes. Rubitecan was isolated by the Stehlin Foundation in the US. SuperGen is currently awaiting regulatory approval in the US and the EU for rubitecan in the treatment of pancreatic cancer. At the BIO-2004 conference, SuperGen announced it is seeking a partner for rubitecan for territories outside the US. SuperGen acquired exclusive worldwide rights to rubitecan from the Stehlin Foundation in 1997 except in Mexico, Canada, Spain, Japan, the UK, France, Italy and Germany. SuperGen has also received approval from the US FDA to use its own manufactured rubitecan in clinical trials. SuperGen and the Stehlin Foundation have an 8-year research agreement that secures global rights to other camptothecins and additional anticancer compounds for the former. In December 1999, SuperGen and Abbott signed a worldwide sales and marketing agreement for rubitecan. Under the terms of the agreement, Abbott had exclusive distribution and promotion rights for rubitecan outside the US, and co-promotion rights with SuperGen within the US. In return, Abbott made an initial equity investment in SuperGen. SuperGen and Abbott Laboratories ended their collaboration agreement in February 2002 by mutual consent with SuperGen stating that the dissolution of the agreement was based on commercial motivation rather than anything to do with rubitecan's safety or efficacy. Abbott no longer has rights or obligations to purchase shares of SuperGen stock or an option to purchase up to 49% of the company. For its part, SuperGen will no longer receive milestone payments worth up to $US57 million. SuperGen has formed a clinical and business alliance with US Oncology (created by the merger between American Oncology Resources and Physician Reliance Network in the US), and will collaborate on clinical trials of rubitecan. SuperGen believes that this relationship will increase the patient population available for trials and enable it to market the drug directly to Oncologists. SuperGen and Capital Research and Management Company have completed a $US16.6 million private placement transaction that will enable future funding for the rubitecan programme as well as other oncology programmes. In July 2004, SuperGen's European subsidiary, EuroGen Pharmaceuticals, submitted a Marketing Authorisation Application for rubitecan in the treatment of pancreatic cancer. The application will be reviewed under the EMEA Centralised Procedure. In June 2003, the EMEA granted SuperGen orphan drug status for rubitecan for the treatment of pancreatic cancer. The US FDA has also granted orphan drug status for rubitecan in the treatment of pancreatic cancer and fast-track status for rubitecan for the treatment of locally advanced or metastatic pancreatic cancer that is resistant or refractory to chemotherapy. SuperGen has conducted three phase III pivotal trials in patients with pancreatic cancer. A phase III randomised trial in chemotherapy-naive patients was conducted at 132 centres throughout the US. The trial enrolled approximately 994 patients who were randomised to receive rubitecan or gemcitabine. Enrollment was completed in October 2001. Another phase III trial has compared rubitecan with the most appropriate chemotherapy in chemotherapy-resistant patients. Enrollment of over 400 patients at 200 medical centres across the US was completed in June 2001. Results from the trial were presented at the 39th Annual Meeting of the American Society of Clinical Oncology (ASCO-2003) [Chicago, US; 31 May - 3 June 2003], after they had been compiled, analysed and submitted to the FDA. The results of the study showed that rubitecan could not help all chemotherapy-resistant patients, but could increase survival in those that do respond. The other phase III pivotal trial was conducted in patients with pancreatic cancer who had failed treatment with gemcitabine. This trial completed enrollment in October 2001, and had enrolled approximately 448 patients. SuperGen is conducting phase II trials of rubitecan in patients with solid tumours in the UK, Italy, France, Germany, the Netherlands and Denmark. Each trial will enroll 100-150 patients with various tumour types, including colorectal, lung, breast, gastric, prostate, cervical and head and neck cancers. Phase I/II trials are underway to investigate rubitecan as a radiosensitiser in patients with lung cancer, and phase II trials in patients with breast cancer are also being conducted. A phase II study in ovarian cancer patients is also being conducted. Results from an ongoing phase II study in cancer patients have shown that rubitecan was effective against chordomas, a rare type of bone cancer. Phase II studies are also underway in haematological malignancies including myelodysplastic syndrome (preleukaemia) and chronic myelomonocytic leukaemia. In February 2000, SuperGen announced that its IND submission for rubitecan had been approved by the Therapeutics Products Programme of Canada. The company stated that it intended to begin clinical trials in Canada in the near future. In February 2004, SuperGen announced an offering of shares of its common stock to finance the commercialisation of rubitecan capsules. In July 2003, SuperGen was granted a US patent covering combination therapies with chemotherapeutic anthracycline agents and structural modifications that may one day lead to next-generation rubitecan compounds. In December 2002, SuperGen was granted US patent No. 6,482,830, covering its polymorphic formulations of rubitecan. The patent also covers a class of polymorphs that are similar to the one at the centre of rubitecan. In addition, SuperGen was also issued US patent No. 6,485,514 in December 2002, covering the local delivery of rubitecan via stents and/or catheters to sites of proliferating cells. Stent- or catheter delivered rubitecan may be beneficial in certain types of cardiac procedures, such as ablation or angioplasty, as well as for direct injection into a certain number of solid tumours. SuperGen is also developing an inhaled, liposomal formulation of rubitecan. It acquired the worldwide rights to this formulation from the Clayton Foundation in December 1999. Inhaled rubitecan is in clinical trials in the US for the treatment of lung cancer and pulmonary metastatic cancer. PMID- 15357632 TI - The phase-dependent photochemical reaction dynamics of halooxides and nitrosyl halides. AB - Recent progress in understanding the phase-dependent reactivity of halooxides and nitrosyl halides is outlined. Halooxide reactivity is represented by the photochemistry of chlorine dioxide (OClO) and dichlorine monoxide (ClOCl). The gas phase photochemical dynamics of OClO are contrasted with the dynamics in condensed environments. The role of excited-state symmetry in defining the reaction dynamics and the observation of photoisomerization resulting in the production of ClOO are discussed. The current understanding of the excited-state reaction dynamics of ClOCl and evidence for photoisomerization of this species resulting in the production of ClClO are outlined. Finally, the photochemical reaction dynamics of the nitrosyl halide ClNO are presented. The main difference between the gas and condensed phase reaction dynamics of this species is that whereas photodissociation to form Cl and NO dominates the gas phase reaction dynamics, photoisomerization resulting in ClON production occurs to an appreciable extent in condensed environments. The observation of photoisomerization for OClO, ClOCl and ClNO suggests that this process is a general feature of the condensed phase reaction dynamics for smaller halooxides and nitrosyl halides. Finally, future areas for study in both halooxide and nitrosyl halide photoreactivity are outlined. PMID- 15357641 TI - Structural characterization of Escherichia coli sensor histidine kinase EnvZ: the periplasmic C-terminal core domain is critical for homodimerization. AB - Escherichia coli EnvZ is a membrane sensor histidine kinase that plays a pivotal role in cell adaptation to changes in extracellular osmolarity. Although the cytoplasmic histidine kinase domain of EnvZ has been extensively studied, both biochemically and structurally, little is known about the structure of its periplasmic domain, which has been implicated in the mechanism underlying its osmosensing function. In the present study, we report the biochemical and biophysical characterization of the periplasmic region of EnvZ (Ala38-Arg162). This region was found to form a dimer in solution, and to consist of two well defined domains: an N-terminal a-helical domain and a C-terminal core domain (Glu83-Arg162) containing both a-helical and b-sheet secondary structures. Our pull-down assays and analytical ultracentrifugation analysis revealed that dimerization of the periplasmic region is highly sensitive to the presence of CHAPS, but relatively insensitive to salt concentration, thus suggesting the significance of hydrophobic interactions between the homodimeric subunits. Periplasmic homodimerization is mediated predominantly by the C-terminal core domain, while a regulatory function may be attributed mainly to the N-terminal a helical domain, whose mutations have been shown previously to produce a high osmolarity phenotype. PMID- 15357643 TI - Hepatocellular carcinoma: role of hepatitis B and hepatitis C viruses proteins in hepatocarcinogenesis. AB - Hepatocellular carcinoma (HCC) is the most important primary hepatic cancer, being a common cancer type worldwide. Many aetiological factors have been related with HCC development, such as cirrhosis, hepatitis viruses and alcohol. Chronic infection with hepatitis B (HBV) and C viruses (HCV) often results in cirrhosis and enhances the probability of developing HCC. The underlying mechanisms that lead to malignant transformation of infected cells, however, remain unclear. HBV is a DNA virus that integrates into the host genome, and this integration is believed, in part, to be carcinogenic. Besides, the virus encodes a 17 kDa protein, HBx, which is known to be a causative agent in the formation of HCC. On the contrary, HCV is a RNA virus that does not integrate into the host genome but likely induces HCC through host protein interactions or via the inflammatory response to the virus. Products encoded in the HCV genome interfere with and disturb intracellular signal transduction. Some HCV proteins, such as the core protein, NS3 and NS5A, have seen to have a regulatory effect on cellular promoters, to interact with a number of cellular proteins, and to be involved in programmed-cell death modulation under certain conditions. The identification of these proteins functions in HCC development and the subsequent development of strategies to inhibit protein-protein interactions may be the first step towards reducing the chronicity and/or of the carcinogenicity of these two viruses. PMID- 15357644 TI - Introduction of NS5A mutations enables subgenomic HCV replicon derived from chimpanzee-infectious HC-J4 isolate to replicate efficiently in Huh-7 cells. AB - Hepatitis C virus (HCV) subgenomic replicon has been reported to replicate efficiently and continuously in human hepatoma Huh-7 cells. To extend the previous results to other isolated HCV clones, we constructed another HCV replicon from HC-J4, one of chimpanzee-infectious HCV clones. An HCV replicon derived from HC-J4 (RpJ4) consists of HCV-5' untranslated region, neomycin phosphotransferase gene, the encephalomyocarditis virus internal ribosomal entry site, HCV nonstructural region, NS3 to NS5B, and HCV-3' untranslated region. The adaptive mutations known to be required for HCV-Con1 replicon were introduced in RpJ4 replicon, aa.(amino acids number according to HC-J4) 2197 serine to proline, deletion of serine at aa.2201, and aa.2204 serine to isoleucine (RpJ4-S2197P, RpJ4-S22001del, and RpJ4-S2204I). RpJ4/ISDR mutant and RpJ4-S2201del/ISDR mutant were also constructed by introducing six amino acid mutations into the interferon sensitivity determining region (ISDR). After transfection into Huh-7 cells and G418 selection, RpJ4 and RpJ4/ISDR mutants did not produce any colony. In contrast, G418-resistant cells were transduced efficiently by RpJ4-S2197P, RpJ4 S2204I, RpJ4-S2201del and RpJ4-S2201del/ISDR mutant, with the RpJ4-S2201del/ISDR mutant being most efficient. Hence the HCV replicon derived from HC-J4 can replicate efficiently following the introduction of adaptive mutations into the upstream region of ISDR. Moreover, additional introduction of mutations into ISDR further enhanced its replication. These findings demonstrate that the genetic structure of the NS5A domain is critical in HCV replications. PMID- 15357645 TI - Interferon-gamma-associated responses to woodchuck hepatitis virus infection in neonatal woodchucks and virus-infected hepatocytes. AB - Acute hepatitis and recovery from woodchuck hepatitis virus (WHV) infection involves increased intrahepatic expression of interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) mRNAs. In the present study, recovery correlated with increased intrahepatic expression of mRNAs for major histocompatibility complex class 1 (MHC1), beta(2)-microglobulin, 2'5' oligoadenylate synthetase (2'5'-OAS), and indoleamine dioxygenase (IDO). By comparison, acute WHV infection progressing to chronicity was associated with diminished expression of these IFN-gamma-associated mRNAs in liver. Transfection of WHV-infected primary hepatocytes (WPH) from WHV carriers with an IFN-gamma expressing plasmid (pIFN-gamma) resulted in dose-dependent accumulations of MHC1, TNF-alpha, 2'5'-OAS, and IDO mRNAs within 96 h. Markers of T cells and immune mediated cytotoxicity that accumulate in recovering liver were not apparent in WPH based on the relative lack of CD3, CD4, Fas ligand, perforin, and granzyme B mRNAs. Expression of pIFN-gamma, and TNF-alpha-expressing plasmid (pTNF-alpha), did not affect total WHV RNA, or fully double-stranded WHV DNA in WPH, but each reduced some of the replicative intermediate (RI) species of WHV DNA synthesis. WPH treated with recombinant IFN-alpha protein had a higher fold induction of 2'5'-OAS mRNA associated with partial reductions in WHV RNAs and the major RI species. Thus, IFN-gamma expression in carrier WPH induced several host responses often observed in liver of recovering woodchucks, and impaired a stage of WHV DNA synthesis by a non-cytolytic mechanism mediated by TNF-alpha. Local enhancement of IFN-gamma-associated responses in chronic WHV-infected hepatocytes may promote therapeutic antiviral effects, but additional effector mechanisms evident during recovery appear necessary for more complete clearance of WHV infection. PMID- 15357646 TI - Racial differences in responses to interferon-beta-1a in chronic hepatitis C unresponsive to interferon-alpha: a better response in Chinese patients. AB - Re-treatment with interferon-alpha alone for chronic hepatitis C nonresponders to interferon-alpha monotherapy is almost ineffective. This multicentre, randomized, parallel-group, dose-finding study evaluated the efficacy of interferon-beta-1a in the treatment of chronic hepatitis C patients unresponsive to interferon alpha. A total of 267 patients were randomized to one of four groups: subcutaneous interferon-beta-1a 12 MIU (44 microg) or 24 MIU (88 microg) administered three times weekly or daily. Patients were treated for 48 weeks and then followed up for an additional 24 weeks. There was a trend towards a dose response relationship regarding virological [loss of detectable serum hepatitis C virus (HCV) RNA] and biochemical response (normalization of serum alanine aminotransferase). Overall, 22 patients (8.3%) had a virological response at the end of treatment; nine patients (3.4%) had a sustained virological response (SVR). Strikingly, 21.7% (5/23) of Chinese patients achieved SVR. Univariate analysis revealed that race was the only variable related to SVR [odds ratio (OR) 16.6; 95% CI 4.1-67.3; P < 0.0001]. Multiple logistic regression analysis also confirmed that more Chinese patients achieved SVR than non-Chinese patients (OR 12.3; 95% CI 2.6-59.3; P = 0.0017). In addition, complete clearance of HCV-RNA occurred earlier in Chinese than in non-Chinese responders (median 2 vs 30 weeks; P = 0.020). Thirty-six patients were withdrawn from treatment because of adverse events. Most adverse events were mild or moderate in severity. In conclusion, interferon-beta-1a provided considerable clinical benefit in Chinese patients with chronic hepatitis C unresponsive to interferon-alpha. The evaluation of interferon-beta-1a in this setting is progressing. PMID- 15357647 TI - The use of lamivudine for patients with acute hepatitis B (a series of cases). AB - There are limited data on the use of lamivudine for patients with severe forms of acute hepatitis B. We report our experience with the use of lamivudine in six patients with acute HBV infection. Lamivudine was justified by disease severity for four patients and by concerns about risk of chronicity for two patients. The diagnoses of the treated patients were: fulminant liver failure (two patients), severe acute hepatitis B, protracted acute hepatitis B, and new HBV infection in the renal dialysis setting (two patients, one with severe liver injury). Serum HBV DNA titres ranged from 10(5) to 10(7) copies/mL prior to commencement of lamivudine. Lamivudine treatment was associated with a decline in serum HBV DNA and serum transaminases in all patients. All but one patient survived. A 58-year old man with fulminant hepatitis and multiple organ failure died despite antiviral treatment. When possible, HBeAg and HBsAg seroconversion was documented during follow-up. In the absence of a randomized, prospective study of lamivudine in patients with severe acute hepatitis B, our data encourage the use of this safe and well tolerated drug. PMID- 15357648 TI - Sustained response after a 2-year course of lamivudine treatment of hepatitis B e antigen-negative chronic hepatitis B. AB - Lamivudine has demonstrated efficacy for the treatment of hepatitis B e antigen negative chronic hepatitis B (e-CHB). However, treatment withdrawal after 1 year has been associated with a high rate of relapse while long-term treatment is associated with increasing risks of drug resistance. We report our treatment experience of 50 Chinese-Canadian patients with e-CHB. All patients received lamivudine for 2 years. Treatment was withdrawn at month 24 in patients who had undetectable hepatitis B virus (HBV) DNA by PCR and normal aminotransferases during the second year of therapy. All patients had HBV genotype B or C. Biochemical response at months 6, 12 and 24 was 74%, 71% and 66%, respectively. HBV DNA was undetectable at months 6, 12 and 24 by hybrid capture and PCR assays in 100%, 92% and 86%; and 94%, 88% and 74% patients, respectively. The cumulative rates of genotypic resistance (GR) after 1 and 2 years were 15% and 25%, respectively. Four (44%) patients with GR experienced a hepatitis flare. The probability of clinical and virological relapse 6, 12, and 18 months after treatment withdrawal were 12% and 30%, 18% and 50%, and 30% and 50%, respectively. Reinstitution of lamivudine resulted in prompt virological and biochemical responses. Our study demonstrates that a sustained response can be achieved after a 2-year course of lamivudine in a subset of patients with e-CHB. PMID- 15357649 TI - High risk of hepatocellular carcinoma in anti-HBe positive liver cirrhosis patients developing lamivudine resistance. AB - The emergence of drug-resistant virus in hepatitis B virus patients treated with lamivudine is well documented. However, its clinical impact in the long-term treatment of anti-HBe positive compensated cirrhotic patients is not well known. In this study, we treated 22 consecutive patients with anti-HBe compensated cirrhosis with lamivudine for a median period of 42 months. All patients responded to lamivudine, but viral breakthrough occurred in 13 patients (59%) between 9 and 42 months of therapy due to the emergence of a mutant strain. During the follow-up, 11 developed hepatocellular carcinoma. Of these, 10 occurred soon after the emergence of viral resistance, generally showing aggressive behaviour, and one in the nine long-term responder patients (P = 0.013). Lamivudine resistance was the only independent predictor of hepatocellular carcinoma development (risk ratio: 10.4; 95% CI: 1.3-84.9). Our study suggests that the occurrence of lamivudine resistance increases the risk of hepatocellular carcinoma in anti-HBe positive cirrhosis and warrants further research. PMID- 15357650 TI - Kinetics of hepatitis B viral load during 48 weeks of treatment with 600 mg vs 100 mg of lamivudine daily. AB - Oral therapy for chronic hepatitis B remains suboptimal. Mathematical modelling of viral decay kinetics to rapidly assess potential antiviral regimens has proved valuable for human immunodeficiency virus and cytomegalovirus. We defined the kinetics of viral replication in 10 chronic hepatitis B patients randomized to lamivudine 100 mg vs 600 mg for 48 weeks. Viral decay kinetics conformed to a biphasic pattern in nine of 10 subjects. Persons receiving 600 mg daily of lamivudine exhibited a 1.6-fold faster decay rate in the infected cell compartment (0.028/day vs 0.017/day, P = 0.06) and a greater overall change in serum viral load when compared with those receiving 100 mg (4.06 vs 1.52 log(10) copies/mL, P = 0.08). More potent therapy appeared to result in more rapid decrease in the infected cell population. Studies using mathematical modelling of viral decay may be a useful method to evaluate single or combination therapy for HBV infection in vivo. PMID- 15357651 TI - Effects of ribavirin combined with interferon-alpha 2b on viral kinetics during first 12 weeks of treatment in patients with hepatitis C virus genotype 1 and high baseline viral loads. AB - This study aimed to find how ribavirin increases viral disappearance in patients with hepatitis C virus (HCV) of genotype 1 and high baseline viral loads (>5.0 x 10(5) copies/mL) when given with interferon (IFN). Using the real-time quantitative polymerase chain reaction, we measured serum HCV in 20 patients during the first 12 weeks of therapy with IFN-alpha 2b and ribavirin. Controls were 10 similar patients given IFN-alpha 2b alone. IFN-alpha 2b was given at 6 MU daily for 2 weeks, and then three times weekly. Ribavirin was given at 600 or 800 mg daily. Serum HCV RNA decreased rapidly in the first phase, during the first 24 h of therapy (day 0), and more slowly in the early second phase (days 1-14). The median decrease was by 1.41 and 0.078 log 10/day in these two phases in the combination therapy group, and 0.90 and 0.081 log 10/day in the monotherapy group. The difference between groups in the first phase was not significant (P = 0.24), nor was that in the next phase (P = 0.68). Later in the second phase, between days 14 and 84, the median decrease was larger in the combination therapy group (0.030 log 10/day) than in the monotherapy group (0.015 log 10/day, P = 0.035). In patients with HCV genotype 1 and high viral loads, the effects of ribavirin with IFN-alpha appeared slowly, after the earliest days of treatment. A long-term favourable outcome of combination therapy may be associated with a rapid viral decline in this later phase of therapy. PMID- 15357652 TI - Different mechanisms of steatosis in hepatitis C virus genotypes 1 and 3 infections. AB - This study reports evidence that hepatocellular steatosis, a frequent histological feature of chronic hepatitis C, is principally metabolic in hepatitis C virus (HCV) genotype 1-infected patients, whereas it is principally virus-induced in HCV genotype 3-infected patients. Multivariate analysis of data on 176 patients with chronic hepatitis C revealed that the severity of steatosis was independently related to HCV RNA load alone in patients infected by HCV genotype 3, whereas it was independently related to the body mass index, daily alcohol intake and histological activity grade (but not viral load) in patients infected by HCV genotype 1. These findings suggest that steatosis is a cytopathic lesion induced by HCV genotype 3, whereas HCV genotype 1 is not steatogenic per se or at the usual in vivo expression levels. PMID- 15357653 TI - Antinuclear antibodies (ANA) in chronic hepatitis C virus infection: correlates of positivity and clinical relevance. AB - We examined correlates of antinuclear antibody (ANA) positivity (ANA+) in individuals with chronic hepatitis C virus (HCV) infection and the effect of positivity on clinical outcome of HCV. Pretreatment sera from 645 patients from three centres in Sweden (n = 225), the UK (n = 207) and Italy (n = 213) were evaluated by indirect immunofluorescence on Hep-2 cells for ANA pattern and titre by a single laboratory. Liver biopsies were all scored by one pathologist. A total of 258 patients were subsequently treated with interferon monotherapy. There was a significant difference in the prevalence of ANA (1:40) by geographic location: Lund 4.4%, London 8.7%, Padova 10.3% [odds ratio (OR) = 0.66; 95% CI: 0.46-0.94; P = 0.023]. Duration of HCV infection, age at infection, current age, route of infection, viral genotype, alcohol consumption, fibrosis stage and inflammatory score were not correlated with ANA+ or ANA pattern. Female gender was correlated with ANA+ and this association persisted in multivariable analyses (OR = 3.0; P = 0.002). Increased plasma cells were observed in the liver biopsies of ANA-positive individuals compared with ANA-negative individuals, while a trend towards decreased lymphoid aggregates was observed [hazard ratio (HR) = 9.0, P = 0.037; HR = 0.291, P = 0.118, respectively]. No correlations were observed between ANA positivity and nonresponse to therapy (OR = 1.4; P = 0.513), although ANA+ was correlated with faster rates of liver fibrosis, this was not statistically significant (OR = 1.8; P = 0.1452). Low titre ANA+ should not be a contraindication for interferon treatment. Our observation of increased plasma cells in ANA+ biopsies might suggest B-cell polyclonal activity with a secondary clinical manifestation of increased serum immunoglobulins. PMID- 15357654 TI - HCV infection and oral lichen planus: a weak association when HCV is endemic. AB - Oral lichen planus (OLP), an immune-mediated disorder, has been reported as an extra-hepatic manifestation of Hepatitis C virus (HCV) infection, especially in HCV hyperendemic areas such as southern Europe and Japan. The aim of this study was to investigate from an epidemiological standpoint whether HCV infection is an important factor affecting the relative risk of OLP in a Mediterranean population or whether this relates to the degree of HCV endemicity. Two cohorts of OLP patients resident in two different regions of southern Italy (Campania and Sicily; n = 859) were evaluated for HCV infection status and categorized into five age classes to respective region-matched controls. No significant difference was found between OLP patients and the general population in this area, when data were corrected by the age-stratified prevalence of HCV. Therefore, the age specific prevalence of HCV infection in OLP patients shows a close trend of direct association with increasing age, without significant differences with the general population of each geographical area. An aetiological link between OLP and HCV cannot be inferred solely by epidemiological data. PMID- 15357655 TI - Hepatitis C virus infection in Egyptian children: single centre experience. AB - The outcome of hepatitis C virus (HCV) infection acquired in childhood is uncertain because of the diversity of the epidemiological and clinical features of infection and disease. The aim of this study was to determine the outcome of HCV infection in 105 Egyptian children who tested positive for HCV antibody (anti HCV). The data of 105 anti-HCV-positive children presenting to the Pediatric Hepatology Unit, Cairo University Children's Hospital, between 1995 and 2002, were retrospectively analysed for risk factors. Seventy-four children with available polymerase chain reaction results were further analysed clinically, serologically and histologically. The age range was 1.3-22 years, with a mean of 11.2 +/- 4.9 years. History of blood transfusion was found in 81 children (77%). HCV RNA was detected in 58.1% of 74 children. Persistently elevated alanine aminotransferase (ALT) levels were present in 40 patients (54.1%). Hepatitis B virus markers (HBsAg and/or anti-HBc) were detected in 18 patients (24.3%). Twenty-six of the 43 HCV RNA-positive children underwent a diagnostic liver biopsy that showed chronic hepatitis in 19 patients (73.1%), cirrhosis in one case only (3.8%), and normal biopsy findings in seven children (26.9%). Blood transfusion remains a major risk of HCV transmission among Egyptian children. HCV infection is not always benign in the childhood period. ALT levels remain elevated in half of the children and histological abnormalities are detected in three quarters of HCV RNA-positive cases. PMID- 15357656 TI - Immune thrombocytopenic purpura in a patient treated with interferon alfacon-1. AB - Immune thrombocytopenic purpura is an acquired disorder characterized by severe thrombocytopenia and caused by one or more antiplatelet autoantibodies. We present a case of a 20-year-old woman referred to our Unit for chronic hepatitis C virus (HCV) infection. At week 28 of treatment with interferon (alfacon-1), undetectable HCV RNA and transaminase levels within normal limits, the patient presented with immune thrombocytopenic purpura, which was successfully treated with immunoglobulin and methylprednisolone. Despite the high doses and long life of corticosteroid treatment HCV RNA remained undetectable. PMID- 15357657 TI - Beneficial effects of melatonin on reperfusion injury in rat sciatic nerve. AB - Studies have shown that ischemia-reperfusion (I/R) produces free radicals leading to lipid peroxidation and to damage of the nervous tissue. Melatonin, a main secretory product of the pineal gland, has free radical scavenging and antioxidant properties and has been shown to diminish I/R injury in many tissues. There are a limited number of studies related to the effects of melatonin on I/R injury in the peripheral nervous system. Therefore, in the present study, the protective effect of melatonin was investigated in rats subjected to 2 hr of sciatic nerve ischemia followed by 3 hr of reperfusion. Following reperfusion, nerve tissue samples were collected for quantitative assessment of malondialdehyde (MDA), an oxidative stress marker, and superoxide dismutase (SOD), a principal antioxidant enzyme. Samples were further evaluated at electron microscopic level to examine the neuropathological changes. I/R elevated the concentration of MDA significantly while there was a reduction at SOD levels. Melatonin treatment reversed the I/R-induced increase and decrease in MDA and SOD levels, respectively. Furthermore, melatonin salvaged the nerve fibers from ischemic degeneration. Histopathologic findings in the samples of melatonin treated animals indicated less edema and less damage to the myelin sheaths and axons than those observed in the control samples. Our results suggest that administration of melatonin protects the sciatic nerve from I/R injury, which may be attributed to its antioxidant property. PMID- 15357658 TI - Protective role of melatonin given either before ischemia or prior to reperfusion on intestinal ischemia-reperfusion damage. AB - Tissue injury resulting from ischemia-reperfusion is of fundamental importance. Experimental evidence suggests that the generation of reactive oxygen species is significantly responsible for this type of injury. In the present study, besides investigating the protective role of melatonin on tissue damage caused by intestinal ischemia-reperfusion, the protective activity of this compound was also analyzed in both pre- and post ischemia melatonin-treated rats. The activities of the main antioxidative enzymes, catalase, superoxide dismutase and glutathione peroxidase in the intestine showed significant (P < 0.05) increases in melatonin-treated animals that were subjected to ischemia/reperfusion compared with those subjected only to ischemia/reperfusion. Also, results clearly indicate that the level of malondialdeyhde, an index of lipid peroxidation, decreased significantly (P < 0.05) when rats subjected to intestinal/reperfusion were given melatonin either before ischemia or before reperfusion. PMID- 15357659 TI - Protective effect of N-acetyl-serotonin on the nonenzymatic lipid peroxidation in rat testicular microsomes and mitochondria. AB - N-acetyl-serotonin, the immediate precursor of melatonin in the tryptophan metabolic pathway in the pineal gland, has been reported to be an antioxidant. The aim of this study was to test the in vitro protective effect of N-acetyl serotonin on the ascorbate-Fe(++) induced lipid peroxidation of polyunsaturated fatty acids (PUFAs) located in testis microsomes and mitochondria. We assayed increasing concentrations (0-10 mM) of N-acetyl-serotonin in testis microsomes and (0-1 mM) of N-acetyl-serotonin in testis mitochondria. Control experiments were performed by incubating microsomal and mitochondrial membranes with N-acetyl serotonin in the absence of lipid peroxidation-inducing drugs. Special attention was paid to the changes produced on the highly PUFAs C20:4 n6 and C22:5 n6. The light emission (chemiluminescence) used as a marker of lipid peroxidation was similar in both organelles when the control and peroxidized groups were compared. N-acetyl-serotonin reduced lipid peroxidation in testicular microsomes or mitochondria for both C20:4 n6 and C22:5 n6. Both long chain PUFAs were protected when N-acetyl-serotonin was incorporated either into microsomes or mitochondria. The N-acetyl-serotonin concentration required to inhibit by approximately 70% lipid peroxidation process was 10 mM in microsomes and between 0.50 and 1 mM in mitochondria. IC 50 values calculated from the inhibition curve of N-acetyl serotonin on the chemiluminescence rates were higher in microsomes (4.50 mM) than in mitochondria (0.25 mM). In these experimental conditions, N-acetyl-serotonin was about 18 times more potent in testicular mitochondria in inhibiting the oxidative processes than it was in testicular microsomes. These results suggest that the protective role of N-acetyl-serotonin in preserving the long PUFAs may be related to its ability to reduce lipid peroxidation. PMID- 15357660 TI - The circadian rhythm of melatonin modulates the severity of caerulein-induced pancreatitis in the rat. AB - Melatonin, an antioxidant, protects the pancreas against acute inflammation but, although this indole is released mainly at night, no study has been undertaken to determine circadian changes of plasma melatonin levels and the severity of acute pancreatitis. The aims of this study were: (a) to compare the severity of caerulein-induced pancreatitis (CIP) produced in the rat during the day and at the night, and (b) to assess the changes of plasma melatonin level and the activity of an antioxidative enzyme; superoxide dismutase (SOD), in the pancreas subjected to CIP during the day time and at night without or with administration of exogenous melatonin or its precursor; l-tryptophan. Rats were kept in 12 hr light/dark cycle. CIP was induced by subcutaneous infusion of caerulein (5 microg/kg/hr for 5 hr). Melatonin (5 or 25 mg/kg) or l-tryptophan (50 or 250 mg/kg) was given intraperitoneally 30 min prior to the start of CIP. CIP induced during the day time was confirmed by histological examination and manifested by pancreatic edema, and rises of amylase and lipase plasma activities (by 400 and 500%, respectively), whereas pancreatic SOD, pancreatic blood flow (PBF) and oxygen consumption by pancreatic tissue (VO(2)) were decreased by 70, 40 and 45%, respectively, as compared with the appropriate controls. All morphological and biochemical parameters of CIP induced at night were significantly less severe, compared with those recorded during the light phase. Plasma melatonin immunoreactivity was significantly higher during the night, than during the day, especially following administration of melatonin or its precursor, which reversed all manifestations of CIP. In conclusion, a circadian rhythm modulates the severity of CIP with a decrease of pancreatitis severity during the night compared with that at the day time and this may be due to the increased plasma level of melatonin and higher activity of SOD in the pancreas. PMID- 15357661 TI - Oxidation of melatonin and its catabolites, N1-acetyl-N2 -formyl-5 methoxykynuramine and N1-acetyl-5-methoxykynuramine, by activated leukocytes. AB - N(1)-acetyl-N(2)-formyl-5-methoxykynuramine (AFMK) and N(1)-acetyl-5 methoxykynuramine (AMK), two melatonin catabolites, have been described as potent antioxidants. We aimed to follow the kinetics of AFMK and AMK formation when melatonin is oxidized by phorbol myristate acetate (PMA) and lipopolysaccharide (LPS)-activated leukocytes. An HPLC-based method was used for AFMK and AMK determination in neutrophil and peripheral blood mononuclear cell cultures supernatants. Samples were separated isocratically on a C18 reverse-phase column using acetonitrile/H(2)O (25:75) as the mobile phase. AFMK was detected by fluorescence (excitation 340 nm and emission 460 nm) and AMK by UV-VIS absorbance (254 nm). Activation of neutrophils and mononuclear cells with PMA produces larger amounts of AFMK than activation with LPS, probably due to the lower levels of reactive oxygen species formation and myeloperoxidase (MPO) degranulation that occurs when cells are stimulated with LPS. The concentration of AMK found in the supernatant was about 5-10% (from 18-hr cultures) compared with AFMK. This result may reflect its reactivity. Indeed AMK, but not AFMK, is easily oxidized by activated neutrophils in a MPO and hydrogen peroxide-dependent reaction. In conclusion, we defined a simple procedure for the determination of AFMK and AMK in biological samples and demonstrated the capacity of leukocytes to oxidize melatonin and AMK. PMID- 15357662 TI - Circadian rhythm entrainment with melatonin, melatonin receptor antagonist S22153 or their combination in mice exposed to constant light. AB - The ability of daily melatonin and the melatonin receptor antagonist, S22153, to entrain circadian system function was investigated in mice with atypical melatonin rhythm. B6D2F(1) mice were first synchronized to a LD 12:12 for approximately 2 wk, then exposed to continuous light (LL) until study completion. After 10-18 days of LL exposure, mice received daily subcutaneous (s.c.) melatonin at a dose of 0.1, 1 or 10 mg/kg/day (exp. 1) or daily intraperitoneal (i.p.) S22153 (20 mg/kg/day) with or without melatonin (1 mg/kg/day, exp. 2) at subjective zeitgeber time (ZT) 10 for 19 days. Then all the mice were exposed to LL for another 10 days. Spectral analysis showed that initial LL lengthened the period of both rhythms by approximately 1.5 hr as compared with LD 12:12. No entrainment of either rhythm was found in controls. Conversely, daily melatonin only, S22153-only or their combination set the temperature and activity periods to approximately 24 hr and produced a significant increase of the circadian amplitude of both rhythms as compared with controls. However, after treatment withdrawal, the dominant period lengthened to approximately 25.5 hr in mice receiving either melatonin or S22153. On the contrary, the period remained close to 24 hr for the 10 days following withdrawal of combined S22153 and melatonin. Such sustained pharmacological resetting of circadian function could display therapeutic potential against external resynchronization resulting from defective photoperiodic entrainment. PMID- 15357663 TI - The endogenous rhythm of plasma melatonin and its regulation by light in the highveld mole-rat (Cryptomys hottentotus pretoriae): a microphthalmic, seasonally breeding rodent. AB - The day- and night-time levels of plasma melatonin were measured in adult male and female highveld mole-rats, Cryptomys hottentotus pretoriae. This study aimed to assess whether melatonin secretion in this nocturnal, strictly subterranean but seasonally breeding rodent has a day-night rhythm and whether that rhythm is circadian and can be modified by photoperiod. In experiment 1, a day-night rhythm of plasma melatonin was found in all animals housed on a 12L:12D schedule, with significantly higher concentrations in the dark (D) compared with the light (L) phase. The increment of plasma melatonin concentration at night was the same on days 1 and 2 for animals in the control group and animals transferred to constant dark. The animals transferred to constant light substantially reduced the amplitude of the melatonin rhythm on day 2. This suggests that the endogenous melatonin rhythm in C. h. pretoriae has a circadian pattern, which can be synchronized by photoperiod and inhibited by exposure to light at night. In experiment 2, the concentration of plasma melatonin in animals kept under 14L:10D (long day, LD) conditions differed significantly from animals on 10L:14D (short day, SD). This finding supports the notion that C. h. pretoriae is sensitive to changes in day length. PMID- 15357664 TI - Melatonin increases cell proliferation in the dentate gyrus of maternally separated rats. AB - Melatonin in mammals, produced by the pineal gland and elsewhere, has shown antioxidant and neuroprotective properties in neuronal cells. We investigated whether melatonin would increase newly born cells (cell proliferation) in the dentate gyrus of maternally separated rats. To examine the effect of melatonin on cell proliferation of the dentate gyrus in maternally separated rats, 5-bromo-2' deoxyuridine (BrdU) immunohistochemistry was performed. Rat pups were separated from their mothers and socially isolated on postnatal day 14. Melatonin (10 mg/kg, i.p.) and BrdU (50 mg/kg, i.p.) were given to them for 7 days. The number of BrdU-positive cells was significantly increased in the dentate gyrus of maternally separated pups with melatonin administration (P < 0.001). In addition, the expression of glucocorticoid receptor was significantly decreased in the dentate gyrus compared with maternally separated pups not given melatonin (P < 0.001). This is the first report that melatonin increases cell proliferation in the dentate gyrus of maternally separated rats. PMID- 15357665 TI - Long-term effects of melatonin or 17 beta-estradiol on improving spatial memory performance in cognitively impaired, ovariectomized adult rats. AB - Melatonin is an endogenously generated potent antioxidant. Our previous studies indicate that melatonin improved learning and memory deficits in APP695 transgenic mouse of Alzheimer's disease. An ovariectomized (OVX) rat model which is characterized by progressive memory deficits, central cholinergic nerve system degeneration and differentiation/apoptosis imbalance is the ideal in vivo model in which to test the neuroprotective effects of melatonin. OVX Sprague-Dawley rats received daily injections of melatonin (5, 10 and 20 mg/kg) or 17 beta estradiol (E2, 80 microg/kg) or sesame oil for 16 wk. Morris water maze results showed that ovarian steroid deprivation resulted in spatial memory impairment, while melatonin and E2 significantly ameliorated spatial memory deficits in OVX rats. The latency to find the hidden platform and the distance to reach the platform become shorter in both melatonin and E2-treated rats compared with those that were only OVX. Four months after OVX, the choline acetyltransferase activity in the frontal cortex and hippocampus were greatly decreased in comparison with the controls. Melatonin and E2 antagonized the effects induced by OVX. Interestingly, the activity of the acetylcholinesterase was not altered in any group of rats. DNA fragmentation was presented in the front cortex of the OVX rats. Melatonin and E2 reduced the number of apoptotic neurons. These findings demonstrate the important effects of melatonin and E2 on cholinergic neurons and support the potential application of melatonin in the treatment of dementia in postmenopausal women. Our results indicate that neuroprotection by melatonin partly correlated to modulation of apoptosis and protection of the cholinergic system. Early long-term melatonin application is a promising strategy which could potentially be applied in a clinic setting. PMID- 15357666 TI - Intracellular second messengers involved in melatonin signal transduction in chicken splenocytes in vitro. AB - The pineal hormone melatonin exhibits immunomodulatory activity well documented in mammals and birds. The mechanism of melatonin action within the immune system is, however, poorly understood. In mammalian immune cells in vitro, melatonin acts mainly as an antiapoptotic, oncostatic and antiproliferative agent, and these effects are exerted via specific receptors or are related to its free radical scavenging activity. In previous studies we have found that in short-term chicken splenocyte cultures in vitro melatonin stimulated basil proliferation and inhibited that stimulated with phytohemagglutinin, a T-cell mitogen. This paper is devoted to the involvement of membrane receptors, previously characterised by us as MT2 (Mel(1b)) and Mel(1c) subtypes, in the above mentioned melatonin effects in chicken splenocyte cultures. For this purpose, in present study a nonselective melatonin receptor antagonist, luzindole, and the selective MT2 blocker, 4P-PDOT, were used. The effect of melatonin on second messengers, cyclic adenosine-3',5'-monophosphate (cAMP) and inositol-1,4,5-trisphosphate (IP(3)), involved in the regulation of proliferation, was examined. We have found that the stimulation of proliferation occurs via Mel(1c) receptor and is associated with the changes in intracellular second messengers concentration: a decrease in cAMP and an increase in IP(3). In contrast, in mitogen-activated splenocytes, melatonin-induced inhibition of proliferation is mediated by MT2 receptors and is related to cAMP accumulation, as well as a decrease in IP(3). In conclusion, we have demonstrated that the stimulatory and inhibitory effect of melatonin on chicken splenocytes in vitro, dependent on the magnitude of cell stimulation, resulted from two different subtypes of membrane receptors. PMID- 15357667 TI - Mechanisms of cancer inhibition by melatonin. PMID- 15357668 TI - NMR solution structure of a potent cyclic nonapeptide inhibitor of ICAM-1 mediated leukocyte adhesion produced by homologous amino acid substitution. AB - We have previously described a disulfide-linked cyclic nonapeptide (inhibitory peptide-01, IP01), with the sequence CLLRMRSIC, which binds to intercellular adhesion molecule-1 (ICAM-1), and blocks binding to its counter-structure, the integrin alphaLbeta2 (leukocyte functional antigen-1, LFA-1) (Sillerud et al., J. Peptide Res. 62, 2003: 97). We now report the optimization of this peptide by means of single homologous amino acid substitutions to yield a new peptide (IP02 K6; CLLRMKSAC) which shows an approximately sixfold improvement in inhibitory activity of multivalent leukocyte binding (inhibition constant for 50% inhibition, IC50 = 90 microm) compared with IP01 (IC50 = 580 microm). This improvement in activity gives IP02-K6 potent in vivo activity in a murine model of ischemia reperfusion injury (Merchant et al., Am. J. Physiol. Heart Circ. 284, 2003: H1260). In order to determine the structural features relevant to ICAM-1 binding, we have determined the structure of IP02-K6 using proton nuclear magnetic resonance (NMR) spectroscopy and restrained molecular modeling. In our previously reported study of solution models of IP01, we observed three interconverting conformations during low-temperature molecular dynamics simulation. In the present study, we find a single conformation of IP02-K6 similar to one of the previously found conformations of IP01 (family C). In particular, an R4-S7 beta-turn is present in similar proportions in both conformation C of IP01 and in IP02-K6; this motif is important in binding to ICAM 1 because this turn enables the IP02-K6 backbone to drape over proline-36 on ICAM 1. The NMR-derived solution model of IP02-K6 was found to dock at the alphaLbeta2 binding site on ICAM-1 with no changes in peptide backbone conformation. This docking model displaced five of the 15 alphaLbeta2 residues at the ICAM-1-binding site and provided a rationale for understanding the quantitative relationship between IP02-K6 structure and biologic activity. PMID- 15357669 TI - Two polymorphs of a covalent complex between papain and a diazomethylketone inhibitor. AB - The three-dimensional structure of two polymorphs of a ZLFG-CH2-papain covalent complex has been determined by X-ray crystallography. The structures indicate that: (i) the methylene carbon atom of the inhibitor is covalently bound to the Sgamma atom of Cys25 of papain; (ii) the hydrophobic S2 pocket formed by Pro68, Val133, Val157, and Asp158 is occupied by the inhibitor's phenylalanyl P2 side chain; (iii) extensive hydrogen bonding and hydrophobic interactions are responsible for the interaction of the inhibitor with the enzyme. Comparison with similar structures suggests that in covalent complexes preservation of main chain main chain interactions between the enzyme and the inhibitor may have higher priority than the P-S interactions. PMID- 15357670 TI - Role of C-terminal heptapeptide in pore-forming activity of antimicrobial agent, gaegurin 4. AB - Gaegurin 4 (GGN4) is an antimicrobial peptide of 37 amino acids isolated from the skin of a frog, Rana rugosa. GGN4 has a disulfide bond between the residues 31 and 37, which is highly conserved among the antimicrobial peptides isolated from skin of the genus, Rana. However, the role of this C-terminal heptapeptide motif is not well understood. In this work, we compared the membrane effects of the full-length GGN4 (C37) and GGN4 1-30 (C30), which is devoid of the C-terminal seven amino acids to elucidate the function of the C-terminal motif. C37 induced significantly larger membrane conductance (>10x) in the model lipid bilayers formed with acidic and neutral phospholipids and larger K+ efflux from gram positive (>30x) and gram-negative bacteria. However, the pores induced by C37 and C30 were not different in their permeability to K+ over Cl- (permeability ratio of K+ to Cl- = 4.8-7.1). In addition, the pore-forming effect of C37 or C30 in acidic membranes was not different from that in neutral membranes. Furthermore, C37-induced K+ efflux was not significantly decreased by the reducing agent, dithiothreitol. The results indicate that C-terminal heptapeptide sequence plays an important role in maintaining the high pore-forming activity of GGN4, but does not participate in forming GGN4-induced pore structure. The disulfide bond in this region does not appear critical for such high ionophoric activity of GGN4. PMID- 15357671 TI - Antimicrobial activity of arginine- and tryptophan-rich hexapeptides: the effects of aromatic clusters, D-amino acid substitution and cyclization. AB - Many antimicrobial peptides bear arginine (R)- and tryptophan (W)-rich sequence motifs. Based on the sequence Ac-RRWWRF-NH2, sets of linear and cyclic peptides were generated by changes in the amino acid sequence, L-D-amino acid exchange and naphthylalanine substituted for tryptophan. Linear RW-peptides displayed moderate activity towards Gram-positive Bacillus subtilis (15 < MIC < 31 microm) and were inactive against Gram-negative Escherichia coli at peptide concentrations < 100 microm. Cyclization induced high antimicrobial activity. The effect of cyclization was most pronounced for peptides with three adjacent aromatic residues. Incorporation of d-amino acid residues had minor influence on the biological activity. The haemolytic activity of all RW-peptides at 100 microm concentration was low (< 7% lysis for linear R/W-rich peptides and < 28% for the cyclic analogues). Introduction of naphthylalanine enhanced the biological activities of both the linear and cyclic peptides. All peptides induced permeabilization of large unilamellar vesicles (LUVs) composed of lipids of the membrane of B. subtilis and erythrocytes, but surprisingly had no effect on LUVs composed of lipids of the E. coli inner membrane. The profiles of peptide activity against B. subtilis and red blood cells correlated with the permeabilizing effects on the corresponding model membranes and were related to hydrophobicity parameters as derived from reversed phase high-performance liquid chromatography (HPLC). The results underlined the importance of amphipathicity as a driving force for cell lytic activity and suggest that conformational constraints and an appropriate position of aromatic residues allowing the formation of hydrophobic clusters are highly favourable for antimicrobial activity and selectivity. PMID- 15357672 TI - Risk factors for oral cancer in newly diagnosed patients aged 45 years and younger: a case-control study in Southern England. AB - BACKGROUND: This case-control study aimed to identify the risk factors for oral cancer in patients aged 45 years and under. METHODS: Patients were recruited over a 3-year period between 1999 and 2001 from 14 hospitals in the southeast of England, UK. RESULTS: Fifty-three (80%) newly diagnosed patients with squamous cell carcinoma (SCC) of the oral cavity participated. The mean age of cases at diagnosis was 38.5 years (SD = 7.0) and 53% were male. Patients were interviewed about main risk factors of tobacco, alcohol, cannabis and their consumption of fresh fruit and vegetables in the past. Ninety-one matched control patients were also recruited. Odds ratios (ORs) and 95% confidence intervals (CI) were obtained from adjusted conditional logistic analyses. Significantly elevated ORs were evidenced amongst males who had started to smoke under the age of 16 years (OR = 14.3; 95% CI: 1.1-178.8). A significant reduction in risk was also shown for ex smokers (OR = 0.2; 95% CI: 0.5-0.8). Consumption of alcohol in excess of recommended amounts also produced an eightfold risk in males (OR = 8.1; 95% CI: 1.6-40.1) and over a fourfold risk of oral cancer from the consumption of excessive amounts of alcohol and having ever smoked (OR = 4.4; 95% CI: 1.1-17.7). CONCLUSION: The study shows that the traditional behavioural risk factors are present in younger people diagnosed with oral cancer. The relatively short duration of exposure and the substantial number of cases without any known risk factors, particularly amongst females, however, suggest that factors other than tobacco and alcohol may also be implicated in the development of oral cancer in a proportion of these younger patients. PMID- 15357673 TI - The expression of NMDA receptor 1 is associated with clinicopathological parameters and prognosis in the oral squamous cell carcinoma. AB - BACKGROUND: Glutamate activates the N-methyl-d-aspartate (NMDA) receptors and this receptor is involved in the proliferation and migration of various tumour cells in vitro. However, the relationship between NMDA receptor expression and clinical parameters in cancer patients is unclear. Therefore, NMDA receptor 1 (NMDAR1) expression along with its clinical significance was examined in patients with oral squamous cell carcinoma (OSCC). METHODS: Eighty-one tumour specimens from OSCC patients were used to determine the NMDAR1 expression level by immunohistochemical staining. The control was obtained from a matched normal adjacent mucosa. The cases were considered to be positive if reactivity was displayed in >25% of the cells. RESULTS: The NMDAR1 reactivity was positive in 50 of 81 cases, while it was negative in the control. NMDAR1 expression was significantly associated with a lymph node metastasis (P = 0.008), the tumour size (P < 0.001), and the cancer stage (P = 0.034). The patients whose tumours expressed NMDAR1 had a significantly poorer survival than the patients who were NMDAR1-negative (log-rank = 6.45, d.f. = 1, P = 0.011). CONCLUSIONS: The NMDAR1 overexpression was significantly associated with the prognosis-related factors. Therefore, it might be one of the prognostic markers of OSCC. PMID- 15357674 TI - The influence of reactivation of the telomerase in tumour tissue on the prognosis of squamous cell carcinomas in the head and neck. AB - BACKGROUND: The reactivation of the telomerase seems to be an important step in the carcinogenesis of most human cancer types. Cell clones, which express this enzyme, get the ability of indefinite proliferation, means become immortal. METHODS: In this study, 80 patients with squamous cell carcinomas (SSC) in oral cavity, oropharynx, hypopharynx and larynx were recorded prospectively concerning a possible correlation of telomerase activity and clinical and prognostic factors. Telomerase activity was analysed by a modified telomeric repeat amplification protocol (TRAP) assay. RESULTS: In 75% of the tumour tissues the telomerase was demonstrated independently of the localization of the tumour. The known clinical prognostic factors did not show any correlation to the expression rate of the telomerase activity in the tumour tissues. Also, reactivated telomerase did not affect the tumour-dependent survival. Only the number of lymph node metastases was in tendency higher in patients with telomerase-positive tumours. The number and timeframe of local and regional recurrences was not influenced by the telomerase status. CONCLUSIONS: Although telomerase seems to be an important part of the carcinogenesis of SCC our data show that the reactivation of telomerase in tumour tissue did not have any prognostic significance for these tumours. The tendency that tumours with active telomerase developed lymph node metastases in a higher number should be evaluated by further enlarged studies for its clinical relevance. PMID- 15357675 TI - The involvement of K(v)3.4 voltage-gated potassium channel in the growth of an oral squamous cell carcinoma cell line. AB - BACKGROUND: In our previous study, an A-type voltage-gated K(+) channel, K(v)3.4, was found more frequently expressed in oral squamous cell carcinoma (OSCC) when compared with non-cancerous matched oral tissue. An OSCC cell line, OECM-1, was found to have moderate level of K(v)3.4 expression. METHODS: To further elucidate the roles of K(v)3.4 for the involvement of neoplastic process, we amplified K(v)3.4 coding sequence by reverse transcriptase polymerase chain reaction (RT PCR), constructed an expression vector carrying this sequence and then stably transfected into OECM-1 OSCC cells. RESULTS: We demonstrated the integration and constitutive expression of K(v)3.4 in the cell. A unique A-type current elicited by such expression in OECM-1 cells was defined by patch clamp analysis. This current pattern can be reversibly blocked by an A-type K(+) channel blocker 2 mM 4-aminopyridine (4-AP). The acquisition of K(v)3.4 activity in OECM-1 cells bestowed growth advantage. However, in 3T3 cell, transfected K(v)3.4 caused only limited increase of growth without forming transformation foci. CONCLUSION: The present study established a stable keratinocyte system carrying functional K(v)3.4 and increase of growth, by which the anti-K(v)3.4 modalities for potential OSCC control can be further investigated. PMID- 15357676 TI - Relative frequency of solitary melanocytic lesions of the oral mucosa. AB - BACKGROUND: Solitary pigmented lesions of melanocytic origin are uncommon in the oral mucosa. These lesions include the oral and labial melanotic macule, oral melanocytic nevus, oral melanoacanthoma, oral melanoma and atypical melanocytic proliferation. The purpose of the study was twofold: to report a large series of solitary melanocytic lesions from one source, and to determine the relative frequency of these lesions. METHODS: The study was based on a systematic search of the files of the Pacific Oral and Maxillofacial Pathology Laboratory, University of the Pacific, San Francisco for solitary pigmented melanocytic lesions (benign and malignant) accessed during the years 1984-2002. RESULTS: Of the 89 430 biopsies accessed during the 19-year period, 773 (0.83%) cases of solitary pigmented melanocytic lesions in the oral mucosa were identified. Oral and labial melanotic macules were the most common melanocytic lesions comprising 86.1% of the entire group and 0.7% of the total number of accessed biopsies. The vermilion border and gingiva were the most common sites (31.1% and 31.0% respectively). Oral melanocytic nevi comprised 11.8% of the entire melanocytic group and 0.1% of the total number of biopsies. The most common site was the palate (44%). Intramucosal nevi were the most common (64%), followed by compound nevi (16.5%) and common blue nevi (16.5%). Junctional nevi were uncommon (3.0%). Oral melanoacanthoma comprised only 0.9% of the entire melanocytic group and 0.008% of the total number of biopsies. Oral melanoma and atypical melanocytic proliferation were the least common lesions each comprising 0.6% of the entire melanocytic group and 0.006% of the total number of biopsies. The most common site for oral melanoma was the palate (60%). CONCLUSION: The palate was the most common location for both melanocytic nevi and oral melanoma. Thus, all melanocytic lesions in the palate should be viewed with caution and biopsy is recommended to rule out melanoma. Further studies are required to elucidate the entity of oral atypical melanocytic proliferation. PMID- 15357677 TI - An exploration of point, annual, and lifetime prevalence in characterizing recurrent aphthous stomatitis in USA children and youths. AB - BACKGROUND: This report presents and contrasts the prevalence of recurrent aphthous stomatitis (RAS) in 51 471 children and adolescents from two major studies: the Third National Health and Nutrition Examination Survey, 1988-1994 (NHANES III) and the National Survey of Oral Health in USA Schoolchildren, 1986 1987 (OHSC), large USA studies based on multistage probability sampling. METHODS: Prevalence proportions, 95% confidence limits and multivariate logistic regression models were constructed for point, 12-month, and lifetime RAS prevalence using SAS-callable SUDAAN 8.0.2. RESULTS: Examinations were performed on 51 471 children and adolescents. Point prevalence was 1.51% (NHANES III) and 1.21% (OHSC); annual prevalence was 19.84% (NHANES III); and lifetime prevalence was 40.18% (OHSC). Multivariate logistic models showed that being white, having a history of herpes labialis, (NHANES III), and being white and an adolescent (OHSC) were predictors of RAS. CONCLUSIONS: Caution should be used in interpreting 12-month and lifetime RAS prevalence based a subject's recall. PMID- 15357678 TI - Increased mast cell density and protease content in actinic cheilitis. AB - BACKGROUND: Actinic cheilitis (AC) is a pre-malignant lesion caused by ultraviolet (UV) radiation and characterized by epithelial and connective tissue alterations. Mast cells (MCs), key contributors to solar elastosis in murine UV irradiated skin, were characterized in order to assess their potential contribution to connective tissue degeneration in AC. METHODS: Actinic cheilitis (n = 15) and normal lip (n = 8) biopsies were stained immunohistochemically for tryptase and enzymehistochemically for chymase to determine MC density and protease content. MC subpopulations (i.e. MC(T) containing only tryptase, and MC(TC) containing chymase and tryptase) and their distribution were also determined. RESULTS: Mast cells and their proteases were increased in AC as compared with normal lip (P < 0.0001), and appeared degranulated especially around elastotic areas. MC(T) predominated over MC(TC) in AC and normal lip (P < 0.05). However, in AC MC(T) were increased in the epithelium/connective junction and connective area (P < 0.05), while in normal lip MC(T) predominated in connective and submucosal areas (P < 0.05). CONCLUSION: The results suggest that increased MC density and protease content may contribute to elastosis formation in AC. In addition, changes in MC(T) distribution may favor AC malignization. PMID- 15357679 TI - Expression patterns of integrins on pleomorphic adenoma and adenoid cystic carcinoma: study on specimens and in vitro investigation of the effects of extracellular matrix on the expression of these adhesion molecules. AB - BACKGROUND: Pleomorphic adenoma (PA) and adenoid cystic carcinoma (ACC) are neoplasms of distinct behaviour, showing similar origin, cell components and marked presence of extracellular matrix (ECM). Interactions between cells and ECM are important in the biology of tumours, being partially mediated by integrins. This study investigated these interactions on PA and ACC using paraffin-embedded tissue and an in vitro model of these conditions. METHODS: Expression of integrins in paraffin-embedded samples was assessed by immunohistochemistry. Cells from PA and ACC were characterized using immunofluorescence, and integrin patterns of expression were investigated on cells cultivated on different ECM proteins. RESULTS: Luminal cells of both PA and ACC were more intensely positive for integrins than myoepithelial cells. In vitro studies revealed that PA cells expressed more integrins than ACC cells regardless the ECM protein present. CONCLUSIONS: This study revealed particular patterns of integrin expression in both specimens and in vitro models of PA and ACC. This might prove useful for a better understanding of the biology of these lesions. PMID- 15357680 TI - Effects of arecoline, safrole, and nicotine on collagen phagocytosis by human buccal mucosal fibroblasts as a possible mechanism for oral submucous fibrosis in Taiwan. AB - BACKGROUND: Oral submucous fibrosis (OSF) is associated with the betel quid chewing habit, and 86% of betel quid chewers in Taiwan are also smokers. Arecoline and safrole are major principles in the composition of betel quid, and nicotine is the main toxic ingredient of cigarettes. METHODS: To explore the pathogenesis of OSF, flow cytometry was used to compare collagen phagocytosis by fibroblasts from the normal and the OSF region of the same 15 OSF patients. RESULTS: The results indicated that heterogeneity of fibroblasts existed because collagen phagocytosis by fibroblasts from the normal region was higher than from the OSF region in the same patient. The percentage of phagocytic cells was significantly inhibited by 10, 25 and 50 microg/ml arecoline, safrole and nicotine in normal fibroblast cultures, respectively, and the percentage of phagocytic cells was significantly reduced by 25, 25 and 50 microg/ml arecoline, safrole and nicotine in OSF fibroblast cultures, respectively. Collagen phagocytosis by fibroblasts exhibited prominent dose-dependent inhibition as the concentration of arecoline, safrole, and nicotine increased. Besides, nicotine had a synergistic effect on arecoline- or safrole-inhibited collagen phagocytosis. CONCLUSIONS: The present study concludes that even in the same person, the collagen phagocytosis by fibroblasts is different between normal and OSF region. The deficiency in collagen phagocytosis by fibroblasts of the lesion might participate in the pathogenesis of OSF. Arecoline, safrole and nicotine, which are released in saliva during BQ chewing plus cigarette smoking, inhibit collagen phagocytosis by fibroblasts in a dose-dependent manner and may induce OSF formation in Taiwan's patients. PMID- 15357682 TI - Mothers' expressed emotion towards children with and without intellectual disabilities. AB - OBJECTIVES: To identify factors associated with maternal expressed emotion (EE) towards their child with intellectual disability (ID). DESIGN AND METHOD: A total of 33 mothers who had a child with ID and at least one child without disabilities between the ages of 4 and 14 years participated in the study. Mothers completed self-assessment questionnaires which addressed their sense of parenting competence, beliefs about child-rearing practices, and their reports of behavioural and emotional problems of their child with ID. Telephone interviews were conducted to assess maternal EE towards the child with ID and towards a sibling using the Five Minute Speech Sample (FMSS; Magana et al. 1986), and also to assess the adaptive behaviour of the child with ID using the Vineland Adaptive Behaviour Scale (VABS; Sparrow et al. 1984). RESULTS: Mothers with high EE towards their child with ID were more satisfied with their parenting ability, and their children had more behaviour problems. Analysis of differential maternal parenting, through comparisons of EE towards their two children, showed that mothers were more negative towards their child with ID for all domains of the FMSS except dissatisfaction. CONCLUSIONS: A small number of factors associated with maternal EE towards children with ID were identified. Differences in maternal EE towards their child with ID and their other child suggest that EE is child-driven rather than a general maternal characteristic. Implications of the data for future research are discussed. PMID- 15357683 TI - Prevention of unintentional injury to people with intellectual disability: a review of the evidence. AB - BACKGROUND: Recent research evidence shows that people with intellectual disability (ID) have double the unintentional injury risk of the general population and the risk is further increased in the presence of psychopathology and epilepsy. The pattern of injury and the circumstances surrounding an injury event in those with ID have some similarity with that of young children in the general population. Interventions to prevent injuries are an important health priority in this vulnerable population. This paper reviews evidence from injury prevention studies for people with ID and also considers the relevance of general population injury interventions for this population. METHOD: Information regarding injury prevention in both ID and general populations was identified using online systems and consultation with research and public health organizations. RESULTS: Few published studies were identified addressing the issue of injury prevention for those with ID. Possible injury prevention strategies appropriate for the major causes of injury in the ID population were identified from the general population literature. While many environmental injury prevention strategies for young children in the general population are applicable to the population with ID, some may require design modification to ensure effectiveness. Other promising approaches include improved information for parents/carers, primary care physician counselling, and home visits by well informed and motivated professionals. There may be injury prevention benefit from improved management of psychopathology and epilepsy. CONCLUSIONS: The issue of injury prevention for those with ID has not been addressed to the extent that the magnitude of the problem requires. Injury prevention programmes trialling a variety of evidence-based approaches and strategies are needed to protect the quality of life for the ID population and their families. PMID- 15357684 TI - Effects of Helicobacter pylori eradication among adults with intellectual disability. AB - BACKGROUND: Compared to the general population, Helicobacter pylori infection is more common among adults with intellectual disability (ID) and is associated with greater levels of disability, maladaptive behaviour, and institutionalization. Little information exists about the effects of eradication therapy in this group, so we aimed to evaluate: (1) success of a standard H. pylori eradication protocol; (2) frequency of side-effects; and (3) impact of eradication on level of functional ability and maladaptive behaviour. METHOD: A cohort of adults with ID underwent assessment of their levels of function and maladaptive behaviour, medical history, physical examination, and H. pylori testing using serology and faecal antigen tests. Some received standard H. pylori eradication therapy. Twelve months later, participants underwent repeat assessment, were grouped by change in H. pylori status and compared. RESULTS: Of 168 participants, 117 (70%) were currently infected with H. pylori at baseline, and 96 (82%) of the 117 were given standard H. pylori eradication therapy. The overall eradication rate was 61% but 31% reported side-effects. Institutional status of the participants, their level of behaviour or function, and number of comorbid medical conditions were not associated with failure of eradication. There were no statistically significant differences in level of behaviour or function, ferritin, or weight between the groups in whom H. pylori was eradicated or stayed positive. CONCLUSION: Adults with ID have lower H. pylori eradication and higher side effect rates than the general population. Levels of maladaptive behaviour and disability did not improve with eradication and thus greater levels of maladaptive behaviour or disability appear to be risk factors for, rather than consequences of, H. pylori infection. PMID- 15357685 TI - Utilization of inpatient care and its determinants among persons with intellectual disabilities in day care centres in Taiwan. AB - BACKGROUND: In Taiwan, current understanding is limited concerning the manner in which health services are utilized by persons with intellectual disabilities (ID). The objective of this study is to describe the patterns of inpatient care sought by persons with ID, and factors affecting inpatient care utilization. METHOD: The primary method used in this study was a cross-sectional survey of 1390 persons with ID in day care centres. Data were obtained from responses to a questionnaire, copies of which were mailed to 30 day care centres catering for persons with ID. The questionnaire assessed demographic and health characteristics, disability status, and inpatient care utilization for the 12 months leading up to the survey. Multivariate logistic regression analysis identified factors independently associated with inpatient care. RESULTS: Findings indicated that the average age of the people with ID in the centres was 13.7 years. Fifty per cent of people were afflicted with multiple handicaps, with an average of 26 outpatient visits made per person during the 12 months, and 16% of persons having been hospitalized within the previous year. The average hospital stay was 6 days. Inpatient care was more likely to be used by those individuals with an ID who were younger, had multiple handicaps, required rehabilitation, and had other disabilities and existing illnesses. CONCLUSIONS: The study concluded that the parameters describing age of persons with ID, as having an existing illness, and requiring rehabilitative care were statistically significant in the logistic regression model of the inpatient care. PMID- 15357686 TI - Mild intellectual disability in children in Lahore, Pakistan: aetiology and risk factors. AB - BACKGROUND: One of the main objectives of studying intellectual disability (ID) in children is to explore its causes. A specific aetiological diagnosis is important in determining the prognosis, nature and extent of services needed to support affected children. METHODS: Aetiology and risk factors in mild ID were studied in a cohort of longitudinally followed children (6-10 years of age, n = 40) in four population groups in and around Lahore, Pakistan. RESULTS: The overall prevalence of mild ID was 6.2%. In 22% of the cases the onset of mild ID was prenatal with small for gestational age and multifactorial inheritance as the main underlying factors. During the postnatal period (28% of the cases), social deprivation and malnutrition were the major causes of ID. In a substantial proportion of the cases (50%), the cause of ID could not be traced. CONCLUSION: The present study indicates a clear relationship of mild ID with prenatal and postnatal malnutrition and social deprivation. Two independent variables, maternal illiteracy and small head circumference at birth, showed a clear association with the development of mild mental disability among children in the study population. PMID- 15357687 TI - An analogue assessment of repetitive hand behaviours in girls and young women with Rett syndrome. AB - BACKGROUND: Rett syndrome is a neurodevelopmental disorder that almost exclusively affects females. In addition to neurodevelopmental regression and loss of hand skills, apraxia, deceleration of head growth, and increasing spasticity and scoliosis, a number of behavioural features are also seen, including stereotypic hand movements, hyperventilation and breath holding. The aim of the study was to investigate the extent to which analogue environmental conditions affected the frequency of repetitive hand behaviour in eight girls and young women with Rett syndrome. METHOD: The frequency of repetitive hand movements was observed every 10 s for four 4-min sessions under the following conditions: Continuous Adult Attention, Adult Demands, Stimulation and No Stimulation. RESULTS: The frequency of repetitive hand movements was high -- they occurred in above 60% of all intervals in all conditions for all participants and at nearly 100% for some participants in some conditions. For one participant the frequency of repetitive hand movements was somewhat reduced in the Stimulation condition; for another it was relatively increased in the No Stimulation condition. CONCLUSIONS: Overall, environmental manipulations had relatively limited effects on repetitive hand behaviours. Repetitive hand behaviour in Rett syndrome may be maintained by automatic reinforcement or neurochemical processes and may not be primarily influenced by contingent reinforcement. PMID- 15357688 TI - Late diagnosis in severe and mild intellectual disability in adulthood. AB - BACKGROUND: The diagnosis of intellectual disability (ID) is highly dependent on a comprehensive personal and family medical history, a complete physical examination and a careful developmental assessment of the patient. Our study intended to: (1) classify the aetiology of mild and severe ID in an adult population of 140 Italian subjects; (2) evaluate the frequency of associated medical conditions; (3) evaluate the age of diagnosis in both groups; and (4) underline the importance of aetiological diagnosis for adult ID patients also. METHODS: The study involved 140 consecutive adult Italian ID inpatients and outpatients neurologically investigated at the Neurological Institute C. Mondino of Pavia Service for Mental Retardation. A total of 80 patients had mild ID (MID group) (39 females, 41 males), mean age 34 years (range 19-61 years), mean IQ = 64 (range 51-75), and 60 had severe ID (SID group) (32 females, 28 males), mean age 30 years (range 19-69 years). They underwent a complete diagnostic work-up that comprised prenatal, perinatal and postnatal history, physical examinations, laboratory investigations, genetic survey and neuroradiological investigations to determine the aetiology of ID and to evaluate the presence of associated medical conditions. RESULTS: ID aetiology was classified as prenatal in 34% of the MID and 28% of the SID group. Perinatal and postnatal events were found in 6% of the MID and in 5% of the SID group. Associated medical conditions were found in 97 patients (47% MID and 26% SID). A genetic diagnosis was possible in 6% of patients above 20 years of age and in 5% of patients above 40 years. A diagnosis of cerebral dysgenesis was possible in 5% of patients above 20 years and 4% of patients above 40 years. CONCLUSIONS: A long interval between the diagnosis of ID and the aetiological definition can be observed in a significant percentage (24%) of our population, leading to unfortunate consequences of late diagnosis: late onset of a specific therapeutic program, genetic counselling that is frequently no more useful, and ineffective prenatal diagnosis, leading to the birth of other affected subjects (for familiar ID). PMID- 15357689 TI - Cross-cultural working in community learning disabilities services: clinical issues, dilemmas and tensions. AB - BACKGROUND: People with learning disabilities from minority ethnic communities face many layers of disadvantage. Providing services that are culturally appropriate and sensitive can be difficult. METHOD: Through the use of clinical examples the issues, tensions and dilemmas raised for a community learning disabilities team in attempting to provide culturally sensitive services in an increasingly multicultural environment are discussed. Ways in which good practice can be more closely achieved are debated. CONCLUSIONS: Conflicts between the principal service values of individuality, choice, promotion of mental health and normalization, whilst also respecting the beliefs and values of other religions and cultures, can raise tensions and dilemmas for learning disabilities services. The importance of sensitive and appropriate service configuration, sharing of good practice, and good staff training are emphasized. PMID- 15357690 TI - Two cases of generalized seizures and the Velocardiofacial syndrome -- a clinically significant association? AB - BACKGROUND: Velocardiofacial syndrome (VCFS) is caused by a micro deletion of chromosome 22q11 and associated with multiple system abnormalities. There is an increasing recognition of associations with psychiatric disorders. Neurological and brain abnormalities have been reported but to date no association with generalized epilepsy has been reported in literature. METHODS: We report two cases of adults with VCFS and primary generalized epilepsy. The details on their background, psychiatric history and history of epilepsy including results of investigations will be discussed. RESULTS: Case 1 is an adult male with mild learning disability (LD), VCFS, paranoid schizophrenia and primary generalized epilepsy. Case 2 is an adult female with mild LD, VCFS, schizoaffective disorder and primary generalized epilepsy. The diagnosis of epilepsy in both cases was based on clinical and EEG findings of primary generalized abnormalities. CONCLUSION: As there is an evidence of association between VCFS and psychiatric disorders, this raises the possibility of an association with generalized epilepsy. PMID- 15357691 TI - Eyewitness testimony and perceived credibility of youth with mild intellectual disability. AB - BACKGROUND: Individuals with intellectual disability (ID) are more vulnerable to abuse compared to individuals without disabilities yet have limited access to the legal system. This study examined perceived credibility of youth with mild intellectual disability (MID) who provide courtroom testimony. METHOD: Participants, 187 undergraduates, were asked general questions about credibility. They also read eyewitness testimony and answered questions about a particular witness's credibility. Half the participants were informed that the youth has MID [chronological age (CA) 15 years, mental age (MA) 10 years] and the others were informed that the youth is a typically developing 10-year-old. RESULTS: When participants were asked general questions about credibility they rated 15-year olds with MID (MA 10 years) as less credible than typically developing 15-year olds and as less credible than typically developing 10-year-olds. However, when participants read eyewitness testimony and answered questions about a particular witness's credibility, no statistically significant differences were found between participants who were informed that the witness was a 15-year-old with MID (MA 10 years) and those who were informed that the witness was a typically developing 10-year-old. CONCLUSIONS: The present study provided a preliminary investigation of perceived credibility of witnesses with MID and suggests directions for future research in this area. PMID- 15357692 TI - Accelerated maturation in brains of patients with Down syndrome. PMID- 15357693 TI - Fish - risks and benefits. PMID- 15357694 TI - Nutritional changes among premenopausal women undertaking a soya based dietary intervention study in Hawaii. AB - BACKGROUND: We conducted a 2-year nutritional intervention among premenopausal women. The goal was to incorporate two daily servings of soya into the regular diet. This report describes the dietary modifications and assesses their nutritional adequacy with regard to major nutrients. METHODS: In this analysis of 100 intervention and 106 control subjects, women completed a validated food frequency questionnaire at baseline; throughout the study, they participated in at least three unannounced 24-h recalls. RESULTS: At randomization, both groups were similar in age and body weight, reported low soya intake, and did not differ by intake of major nutrients and foods. According to the 24-h recalls, women in the intervention group consumed nearly two servings of soya per day, while the control women remained at 0.2 servings. In comparison with the control group, the intervention group consumed fewer dairy products, primarily milk, but also less meat, nuts and seeds. As a result of the dietary modification, the intervention women consumed less-saturated fat and cholesterol and more protein, dietary fibre, calcium and vitamins than the control group. CONCLUSION: These results suggest that women in the intervention group improved the overall quality of their diet by adding two servings of soya per day. PMID- 15357696 TI - Effects of a sweet and a nonsweet lunch on short-term appetite: differences in female high and low consumers of sweet/low-energy beverages. AB - INTRODUCTION: Effects of sweet taste on short-term appetite are still being actively researched. This study investigates the proposal that the effects of sweet tastes on appetite may differ as a result of differing habitual experiences of sweetness with or without energy. METHODS: Effects of sweet tastes on appetite were investigated in habitual high and low consumers of sweet/low-energy beverages. Sweet taste was manipulated in a preload lunch and appetite was subsequently measured using test meal intake and subjective ratings of general and specific appetites. RESULTS: The effects of the sweet and nonsweet lunch on short-term appetite differed significantly in high and low consumers of sweet/low energy beverages, in subjective ratings of appetite for something sweet [consumer x preload x time interaction F(12,126) = 2.68, P = 0.003] and appetite for something savoury [consumer x preload x time interaction F(12,126) = 3.17, P = 0.001]. Effects in low consumers of sweetness without energy demonstrate close association between taste and energy, whereas effects in high consumers suggest a dissociation between taste and energy in these consumers. DISCUSSION: These findings provide a further indication that the short-term control of appetite varies according to the habitual pattern of dietary intake. The long-term experience of sweetness without energy influences appetite for sweet and savoury tastes. PMID- 15357697 TI - Audit of successful weight maintenance in adult and paediatric survivors of thermal injury at a UK regional burn centre. AB - BACKGROUND: Thermal injury is associated with significant weight loss, particularly of lean body mass. Weight loss increases the risk of morbidity and increased length of stay (LOS). Appropriate nutrition support can improve these outcomes. The aim of this audit was to evaluate the success of nutrition intervention in minimizing weight loss in adults and children with thermal injury. METHOD: Patients referred for nutrition support were monitored for weight change during their inpatient care episode. Sixty cases met the study criteria. Admission and discharge weight profiles were identified. The degree of weight loss was calculated. Comparison was made to a recognized standard of acceptable weight loss. RESULTS: On admission five (13.2%) adults were found to be clinically underweight. All children were within the acceptable weight percentile range. Adults demonstrated higher (mean +/- SD) percentage weight loss (8.1 +/- 6.0%) than children (4.7 +/- 4.5%). About 76.7% of all cases met the standard used. More children (95.5%) met the standard than adults (65.8%). CONCLUSIONS: Most subjects maintained weight within an acceptable limit during the inpatient episode. Children appeared particularly successful at weight maintenance. Reasons are multifactorial and warrant further investigation. PMID- 15357698 TI - Maternal - neonatal folate and vitamin B12 serum concentrations in Greeks and in Albanian immigrants. AB - BACKGROUND: Low folate and vitamin B(12) concentrations during pregnancy are implicated with neural tube defects (NTD) and neurological manifestations in the neonates. AIM: To compare serum concentrations of folate and vitamin B(12) in 1025 Greek mothers, 908 immigrant Albanian mothers at delivery and in the cord blood of their neonates. METHODS: A 30 days food diary was kept from each mother. Folate and vitamin B(12) sera concentrations were measured with Bayer ADVIA Centaur System. RESULTS: Energy, carbohydrates and total fat intake were significantly higher in Albanian mothers when compared with those of Greek origin. On the contrary, total protein, and especially animal protein intake were higher in the Greek mothers. Folate intake was similar between the two ethnic groups whereas vitamin B(12) intake in Albanians was evaluated lower (1.8 +/- 0.13 microg day(-1)) when compared with that of Greeks (2.8 +/- 0.12 microg day( 1); P < 0.001). Folate serum concentrations in mothers and newborns were similar (17.7 +/- 9.1 and 26.5 +/- 15.2 nm versus 18.1 +/- 8.6 and 24.6 +/- 14.7 nm, respectively P > 0.05) in the two ethnic groups. In contrast, vitamin B(12) serum concentration was significantly lower (135.0 +/- 19.6 pm) in the Albanian mothers compared with that of Greeks (164.5 +/- 17.7 pm, P < 0.0001). Consequently, the vitamin concentrations was found lower in the newborns of Albanian origin (133.6 +/- 11.8 pm) compared with that of Greek neonates (213.3 +/- 11.4 pm, P < 0.0001). CONCLUSIONS: (a) Folate serum concentrations in Greek and Albanian mothers and in their neonates were similar, (b) vitamin B(12) serum concentrations, evaluated for first time in these ethnic groups, were significantly low in Albanians, both in mothers and newborns, (c) the low protein and especially the reduced animal protein intake from the Albanian mothers, possibly due to their low socio-economic status, may be responsible for their decreased vitamin B(12) serum concentrations and (d) vitamin B(12) supplementation along with an increase of animal protein intake might improve the vitamin serum status in the Albanians. PMID- 15357699 TI - The health benefits of omega-3 polyunsaturated fatty acids: a review of the evidence. AB - The UK dietary guidelines for cardiovascular disease acknowledge the importance of long-chain omega-3 polyunsaturated fatty acids (PUFA) - a component of fish oils - in reducing heart disease risk. At the time, it was recommended that the average n-3 PUFA intake should be increased from 0.1 to 0.2 g day(-1). However, since the publication of these guidelines, a plethora of evidence relating to the beneficial effects of n-3 PUFAs, in areas other than heart disease, has emerged. The majority of intervention studies, which found associations between various conditions and the intake of fish oils or their derivatives, used n-3 intakes well above the 0.2 g day(-1) recommended by Committee on Medical Aspects of Food Policy (COMA). Furthermore, in 2004, the Food Standards Agency changed its advice on oil-rich fish creating a discrepancy between the levels of n-3 PUFA implied by the new advice and the 1994 COMA guideline. This review will examine published evidence from observational and intervention studies relating to the health effects of n-3 PUFAs, and discuss whether the current UK recommendation for long chain n-3 PUFA needs to be revisited. PMID- 15357700 TI - Inter-brand differences in iron content of breakfast cereals and their impact on measurement of iron intake. AB - BACKGROUND/AIMS: Fortified breakfast cereals (FBCs) are an important source of iron in the UK diet. In order to quantify their contribution to iron nutrition, food composition data for these products must be accurate. The very large amount of products available, together with inter-brand differences in iron content mean that discrepancies between the iron content of many FBCs and values in standard food composition databases (FCD) exist. The variation in reported iron contents of FBCs was examined and the impact of this variation on measurement of iron intake using standard food composition tables was investigated. METHOD: Data on the reported iron content of 128 FBCs were collected. Completed food diaries from 291 participants of the UK Women's Cohort study were used in the analysis. Mean iron intake from a 4-day food diary was calculated using UK food tables' values for FBCs. This was repeated using values reported by the manufacturer for each brand of cereal. The two sets of results were then compared. RESULTS: There is wide variation in iron content of breakfast cereals available in the United Kingdom. Use of FCD values instead of brand-specific values under- or overestimates an individual's iron intake by as much as 28 or 22% respectively. These results suggest that use of FCD values for breakfast cereals is potentially a source of substantial error in measurement of individuals' iron intake. CONCLUSION: Dietitians need to be aware of inter-brand differences in iron content and formulate advice accordingly. Failure to collect brand-specific data for the iron content of FBCs could lead to measurement error in measuring iron intake in dietary studies. PMID- 15357701 TI - Validation of energy intake by dietary recall against different methods to assess energy expenditure. AB - OBJECTIVES: To compare the validity of dietary recalls and physical activity recalls and investigate some factors influencing this validity. To provide an example showing how procedures based on recalls of physical activity can assess the validity of dietary recalls and identify subjects constantly underreporting their energy intake (EI). DESIGN AND SUBJECTS: Thirty-seven women were studied using three 24-h dietary recalls, two kinds of physical activity recalls, indirect calorimetry and the doubly labelled water method. RESULTS: The EI obtained using dietary recalls were biased with respect to body mass index (BMI) and attitudes towards body weight and dieting, whereas results obtained using a physical activity recall were not. Eighteen women produced underreports (UR), i.e. their average EI was below 76% of total energy expenditure (TEE), whereas 24 women reported an EI that was lower than TEE on all three recall days, i.e. constantly underreporting subjects. A physical activity recall identified 13 URs and 20 of the constantly underreporting subjects. CONCLUSIONS: In contrast to estimates of EI, TEE assessed using physical activity recalls was not biased with respect to BMI or attitudes towards body weight and dieting. Recalls of physical activity represent potentially useful procedures for identifying URs and constantly underreporting subjects but are not accurate enough for individuals. PMID- 15357708 TI - Bacterial enteritis and the development of the larval digestive tract in olive flounder, Paralichthys olivaceus (Temminck & Schlegel). AB - Three bacterial isolates obtained from diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization and 16S rRNA gene sequencing studies. Bacterial enteritis was reproduced in 16 and 22 days post-hatch (dph) larvae by administering brine shrimp nauplii, Artemia salina, dosed with the environmental isolates and reference strains of V. ichthyoenteri. To investigate the effect of the disease on development of the stomach, a pepsin activity assay and reverse transcription-polymerase chain reaction (RT-PCR) analysis of the expression of the pepsinogen gene were performed. Expression of olive flounder pepsinogen was detected from 30-dph larvae and the increased level of pepsin activity coincided with reduced susceptibility to the disease. Growth rates of V. ichthyoenteri, V. anguillarum and Edwardsiella tarda were tested in artificial stomach conditions using HCl and porcine pepsin. All the strains of V. ichthyoenteri were inhibited by low pH conditions which corresponded with an increase in pepsin levels. This suggests that differentiation of the stomach in olive flounder larvae and juveniles, an essential physiological development, also provides the host with a non-immunological defence mechanism. PMID- 15357709 TI - Fusarium incarnatum isolated from black tiger shrimp, Penaeus monodon Fabricius, with black gill disease cultured in Vietnam. AB - Fusarium incarnatum was isolated from gill lesions of cultured black tiger shrimp, Penaeus monodon, in every crop during 2000-2002 in Nghe An province, Vietnam. Infected shrimps showed typical signs of black gill disease and mortalities about a month prior to harvest. Detailed morphological examinations, as well as molecular phylogenic analyses based on partial nucleotide sequences of ribosomal DNA, were made on the isolates. An artificial infection of kuruma prawn, Penaeus japonicus, using two selected isolates was also conducted and their pathogenicity determined. PMID- 15357710 TI - Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans. AB - The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp. PMID- 15357711 TI - Skeletal myopathy in juvenile barramundi, Lates calcarifer (Bloch), cultured in potassium-deficient saline groundwater. AB - Saline groundwater is being pumped from a number of locations in rural Western Australia to prevent secondary salinity impacting farmland, rural infrastructure and areas with high conservation value. Aquaculture may offset the costs of groundwater pumping, and the suitability of groundwater for finfish aquaculture is being assessed through bioassays. There are marked spatial variations in the ionic composition of saline ground water in Western Australia and this paper describes two bioassays investigating a saline, potassium-deficient water source that resulted in mortalities in juvenile barramundi, Lates calcarifer (Bloch). Histopathological examination revealed severe degeneration and necrosis of skeletal muscles, marked hyperplasia of branchial chloride cells and renal tubular necrosis. Clinical chemistry findings included hypernatraemia and hyperchloridaemia of the blood plasma and lowered muscle potassium levels. It is concluded that the principal cause of death was skeletal myopathy induced by low water potassium levels. PMID- 15357712 TI - Histological alterations in the liver of sea bream, Sparus aurata L., caused by short- or long-term feeding with vegetable oils. Recovery of normal morphology after feeding fish oil as the sole lipid source. AB - This study evaluated the effects of fish oil (FO) replacement by vegetable oils [soybean oil (SO), rapeseed oil (RO), linseed oil (LO)] and subsequent feeding with FO on the liver morphology of sea bream. A short-term trial (3 months) and long-term trial (6 months) were carried out feeding sea bream with the following experimental diets: FO100%; SO60% + FO40%; RO60% +FO40%; LO60% + FO40%; SO + RO +LO60% + FO40%. Finally, all groups from the long-term trial were fed with FO100% for 95 days (washout period). Liver samples were taken for histological and biochemical studies. In both the short- and long-term trials, livers of sea bream fed LO60% and SO + RO + LO60% showed a similar hepatic morphology to that observed in fish fed FO100%. In contrast, sea bream fed SO60% showed an intense steatosis, with foci of swollen hepatocytes containing numerous lipid vacuoles. After the washout period, a considerable reduction of the cytoplasmic vacuolation and the lipid vacuole accumulation were observed in the livers of fish fed the different experimental diets. The results of this study suggested that the type of non-essential fatty acid, characteristic of vegetable oils, induces the appearance of steatosis in the following order: linoleic acid > linolenic acid > oleic acid. However, the liver alterations found during the experimental periods with vegetable oils are reversible when the fish are re-fed with a balanced diet (FO100%), indicating the non-pathological character of these histological changes. PMID- 15357713 TI - First observation of herpes-like virus particles in northern pike, Esox lucius L., associated with bluespot-like disease in Ireland. AB - Circular whitish granular lesions, 5-12 mm in diameter, were observed on the skin and fins of a wild northern pike, Esox lucius, caught in a lake in the Republic of Ireland. Histological examination of the lesions revealed hypertrophied cells in the epidermis with deeply basophilic enlarged nuclei and dark-staining granular material in the cytoplasm. Transmission electron microscopy of these cells revealed naked hexagonal herpes-like virus nucleocapsids (97 +/- 7 nm) in their nuclei while the cytoplasm contained multiple aggregates of enveloped viral particles. This is the first report of herpes-like virus particles in northern pike originating outside North America, where esocid herpesvirus-1 (EsHV-1) has previously been reported. Shared clinical, histological, morphological and epidemiological findings suggest that the observed particles in this report may also be EsHV-1. PMID- 15357714 TI - Remodelling of mixed bone during infections of Myxobolus scleroperca (Myxozoa) in Percina caprodes (Rafinesque) (Percidae) from the St Lawrence River, Quebec. PMID- 15357715 TI - The ability of two different Vibrio spp. bacteriophages to infect Vibrio harveyi, Vibrio cholerae and Vibrio mimicus. AB - AIMS: To determine the host range of the Vibrio harveyi myovirus-like bacteriophage (VHML) and the cholera toxin conversion bacteriophage (CTX Phi) within a range of Vibrio cholerae and V. mimicus and V. harveyi, V. cholerae and V. mimicus isolates respectively. METHODS AND RESULTS: Three V. harveyi, eight V. cholerae and five V. mimicus isolates were incubated with VHML and CTX Phi. Polymerase chain reaction (PCR) was used to determine the presence of VHML and CTX Phi in infected isolates. We demonstrated that it was possible to infect one isolate of V. cholerae (isolate ACM #2773/ATCC #14035) with VHML. This isolate successfully incorporated VHML into its genome as evident by positive PCR amplification of the sequence coding part of the tail sheath of VHML. Attempts to infect all other V. cholerae and V. mimicus isolates with VHML were unsuccessful. Attempts to infect V. cholerae non-01, V. harveyi and V. mimicus isolates with CTX Phi were unsuccessful. CONCLUSIONS: Bacteriophage infection is limited by bacteriophage-exclusion systems operating within bacterial strains and these systems appear to be highly selective. One system may allow the co-existence of one bacteriophage while excluding another. VHML appears to have a narrow host range which may be related to a common receptor protein in such strains. The lack of the vibrio pathogenicity island bacteriophage (VPI Phi) in the isolates used in this study may explain why infections with CTX Phi were unsuccessful. SIGNIFICANCE AND IMPACT OF THE STUDY: The current study has demonstrated that Vibrio spp. bacteriophages may infect other Vibrio spp. PMID- 15357716 TI - Role of viability of probiotic strains in their persistence in the gut and in mucosal immune stimulation. AB - AIMS: To determine how probiotic bacteria contact with intestinal epithelial and immune cells and the conditions to induce a good mucosal immune stimulation. METHODS AND RESULTS: Lactobacillus casei was studied by transmission electron microscopy (TEM) to determine its interaction with the gut. We compared the influence of viable and nonviable lactic acid bacteria on the intestinal mucosal immune system (IMIS) and their persistence in the gut of mice. TEM showed whole Lact. casei adhered to the villi; the bacterial antigen was found in the cytoplasm of the enterocytes. Viable bacteria stimulated the IMIS to a greater extent than nonviable bacteria with the exception of Lact. delbrueckii subsp. bulgaricus. For all the strains assayed at 72 h no antigenic particles were found in the intestine. CONCLUSION: Antigenic particles but not the whole bacteria can enter to epithelial cells and contact with the immune cells. Bacterial viability is a condition for a better stimulation of the IMIS. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrated that only antigenic particle interact with the immune cells and their fast clearance from the gut agrees with those described for the particulate antigens. The regular consumption of probiotics should not adversely affect the host. PMID- 15357717 TI - Microbial characteristics of marine sediments in bathing area along Pesaro Gabicce coast (Italy): a preliminary study. AB - AIMS: This paper evaluates the presence of human pathogen micro-organisms in marine sediments in a coastal area suitable for bathing. In addition, the grain size analysis was performed in order to correlate the characteristics of the sediments and the microbial content. METHODS AND RESULTS: The samples were collected in two small bays along the central Adriatic coast, where breakwaters had been built for the purpose of halting marine erosion. Faecal contamination indicators, Salmonella and Vibrio species, enteric viruses were investigated using standard techniques for isolation and identification. The grain size was determined using calibrated sieves and 'Sedigraph' device. In some samples, the faecal contamination indices overstepped legislative limits. Salmonella was never found. Vibrio and enteric viruses were isolated: the micro-organisms were preferentially abundant in fine sediments. CONCLUSIONS: Marine sediments can represent an important reservoir of allochthonous and marine micro-organisms and the microbial charge correlates with the characteristics of the sediments. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that it is important to analyse marine sediments before defining the quality of coastal areas. PMID- 15357718 TI - Detection of Helicobacter pylori by PCR but not culture in water and biofilm samples from drinking water distribution systems in England. AB - AIMS: To investigate treated water distribution systems in England as a source of Helicobacter pylori. METHODS AND RESULTS: Water and biofilms were obtained from 11 domestic and seven educational properties and from hydrants, reservoirs and water meters supplied by three water utilities. Samples were cultured on nonselective and antibiotic containing media combined with immunomagnetic separation concentration. Viable helicobacters were not detected in any of the 151 samples but Helicobacter-specific PCR assays detected DNA in 26% of samples from domestic properties, schools and hydrants with the highest frequency in biofilms (42%). Direct sequencing of six selected amplicons confirmed >95% sequence homology to H. pylori. CONCLUSIONS: While viable helicobacters were not isolated, evidence was obtained for the presence of Helicobacter DNA, including that of H. pylori. Biofilms on surfaces within water distribution systems may act either as sites for the passive accumulation of helicobacters or as potentially important reservoirs of infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings strengthen evidence that H. pylori may be transmitted through drinking water. However, there is currently no evidence that viable cells can survive the disinfection levels used in UK mains supplies and the health risk from this source remains unclear. PMID- 15357719 TI - Comparative analysis of antibiotic and antimicrobial biocide susceptibility data in clinical isolates of methicillin-sensitive Staphylococcus aureus, methicillin resistant Staphylococcus aureus and Pseudomonas aeruginosa between 1989 and 2000. AB - AIMS: To analyse population minimum inhibitory concentrations (MICs) data from clinical strains of Staphylococcus aureus and Pseudomonas aeruginosa for changes over a 10-year period and to look for correlations between the antimicrobials tested. METHODS AND RESULTS: Data from the MIC study of 256 clinical isolates of Staph. aureus [169 methicillin-sensitive Staph. aureus (MSSA), 87 methicillin resistant Staph. aureus (MRSA)] and 111 clinical isolates of Ps. aeruginosa against eight antimicrobial biocides and several clinically relevant antibiotics was analysed using anova, Spearman-Rho correlation and principal component analysis. Comparisons suggest that alterations in the mean susceptibility of Staph. aureus to antimicrobial biocides have occurred between 1989 and 2000, but that these changes were mirrored in MSSA and MRSA suggests that methicillin resistance has little to do with these changes. Between 1989 and 2000 a sub population of MRSA has acquired a higher resistance to biocides, but this has not altered the antibiotic susceptibility of that group. In both Staph. aureus and Ps. aeruginosa several correlations (both positive and negative) between antibiotics and antimicrobial biocides were found. CONCLUSIONS: From the analyses of these clinical isolates it is very difficult to support a hypothesis that increased biocide resistance is a cause of increased antibiotic resistance either in Staph. aureus or in Ps. aeruginosa. SIGNIFICANCE AND IMPACT OF THE STUDY: The observation of negative correlations between antibiotics and biocides may be a useful reason for the continued use of biocides promoting hygiene in the hospital environment. PMID- 15357720 TI - Genetic marker for differentiating beer-spoilage ability of Lactobacillus paracollinoides strains. AB - AIMS: To determine whether the beer-spoilage ability is an intrinsic character of Lactobacillus paracollinoides and identify a genetic marker for differentiating the beer-spoilage ability of strains belonging to this species. METHODS AND RESULTS: The ribotype of a nonspoilage strain, Lact. brevis ATCC8291, was found to be identical with that of Lact. paracollinoides LA7. The 16S rDNA sequence analysis and DNA-DNA hybridization study indicates that nonspoilage ATCC8291 should belong to Lact. paracollinoides. We further isolated nonspoilage variants from Lact. paracollinoides LA2(T) and LA9 by incubating these strains at 30 degrees C. To identify a genetic marker for differentiating the beer-spoilage ability of Lact. paracollinoides, open reading frames 5 (ORF5), the previously reported genetic marker for Lact. brevis, was evaluated. As a result, ORF5 homologues were detected in all of the 12 beer-spoilage strains of Lact. paracollinoides, while this ORF was not found in ATCC8291 or the two nonspoilage variants obtained from LA2(T) and LA9. CONCLUSIONS: Lactobacillus paracollinoides is not an intrinsic beer-spoiler and the nonspoilage strain Lact. brevis ATCC8291 should be reclassified as Lact. paracollinoides. ORF5 was found to be useful for differentiating beer-spoilage ability of this species. SIGNIFICANCE AND IMPACT OF THE STUDY: The finding that Lact. paracollinoides includes nonspoilage strains necessitates brewers to use a genetic marker that is associated with the beer spoilage ability of this species. PMID- 15357721 TI - PCR detection of staphylococcal enterotoxin genes in Staphylococcus spp. strains isolated from meat and dairy products. Evidence for new variants of seG and seI in S. aureus AB-8802. AB - AIMS: Evaluation of the occurrence of most known staphylococcal enterotoxin (SE) genes, egc (enterotoxin gene cluster) and TSST1 (toxic shock syndrome toxin 1) gene in both coagulase-positive (CPS) and coagulase-negative (CNS) staphylococcal strains isolated from meat and dairy products. METHODS AND RESULTS: Specificity and reliability of the PCR detection methods used were ascertained by using nine reference strains of Staphylococcus (S. aureus) harbouring SE genes (seA to seE; seG, seH, seI, seM, seJ, seN and seO) and egc (containing the following sequence of genes: seO, seM, seI, phient1, phient2, seN and seG). Of 109 wild Staphylococcus spp. strains analysed, only 11 S. aureus strains were SE and/or TSST1 PCR-positive. The last 11 strains also appeared to harbour the egc. Restriction endonuclease analysis of part of the egc of both reference and wild strains showed that different variants of the egc exist. Moreover, nucleotide sequences of seG and seI indicate that the egc of the strain AB-8802 is characterized by the presence of variants of these enterotoxins (seGv and seIv). CONCLUSIONS: The occurrence of SE genes in CNS and other non-S. aureus species isolated from Napoli-type salami, raw water buffalo milk and natural whey cultures used for mozzarella cheese manufacturing is very rare. SIGNIFICANCE AND IMPACT OF THE STUDY: During this study it was shown that at least five different egc may exist in S. aureus. A thorough study of egc polymorphism should provide further insight into the phylogenetics of the egc. PMID- 15357722 TI - Antimicrobial activity of enterocin EJ97 against 'Bacillus macroides/Bacillus maroccanus' isolated from zucchini puree. AB - AIMS: Activity of the bacteriocin EJ97 produced by Enterococcus faecalis EJ97 against strains of 'Bacillus macroides/B. maroccanus' isolated from spoiled zucchini puree was investigated. METHODS AND RESULTS: The influence of several factors like bacteriocin concentration, incubation temperature, pH, growth medium and chemical perservatives on bacteriocin activity was investigated. Enterocin EJ97 [2 arbitrary units (AU) per millilitre] had a marked bactericidal effect on strain INRA P53-2 after 4 h of incubation at 37 degrees C, 24 h at 15 degrees C or 48 h at 4 degrees C. Activity was markedly reduced at pH values of 5.0 and 9.0, but was potentiated by sodium nitrite, sodium benzoate, sodium lactate and sodium tripolyphosphate. Inhibition of strain INRA P53-2 in a commercial vegetable puree required a 10-fold higher bacteriocin concentration. Strain EJ97 was able to grow and produce bacteriocin on vegetable puree, but no inhibition of strain INRA P53-2 was detected. CONCLUSIONS: The concentration-dependent bactericidal activity of enterocin EJ97 against strain INRA P53-2 was higher at 37 degrees C and neutral pH, and was potentiated by chemical preservatives. Although enterocin EJ97 was less active in vegetable puree, the concentrations providing bactericidal activity in this food matrix are practical for commercial use. SIGNIFICANCE AND IMPACT OF THE STUDY: Enterocin EJ97 may have a potential for use in the prevention of food spoilage caused by 'B. macroides/B. maroccanus'. PMID- 15357723 TI - Liquid chromatography/mass spectrometry analysis of small-scale glycopeptidolipid preparations to identify serovars of Mycobacterium avium-intracellulare complex. AB - AIMS: The antigenic glycopeptidolipids (GPLs) from Mycobacterium avium intracellulare complex (MAC) are grouped into 28 serovars on the basis of the variable oligosaccharide sequences and the core structures. To facilitate the identification of MAC serovars by employing liquid chromatography/mass spectrometry (LC/MS), the diversity in fatty acyl moieties and the number of acetyl groups of GPLs should be characterized. METHODS AND RESULTS: Employing a small-scale preparation method, sufficient quantities of intact GPLs could be obtained from several colonies of MAC within 4 h. Tandem mass spectrometry of GPLs showed the presence of common fragment ion at m/z 1048 in the main molecular species of all reference strains. It revealed that the acyl moieties had similar diversity among all serovars. Furthermore, intact GPLs had mainly one or two acetyl groups. This allowed us to determine the masses of each serovar based on intact GPLs and to classify 16 isolates from patients by LC/MS. CONCLUSIONS: The present serotyping method using LC/MS analysis improved the precision of measurements and shortened the procedure time compared with conventional thin layer chromatography or the seroagglutination test method. SIGNIFICANCE AND IMPACT OF THE STUDY: This proposed method proves useful for identifying serovars of MAC for epidemiological and pathogenic research purposes. PMID- 15357724 TI - The aggregation-promoting factor of Lactobacillus crispatus M247 and its genetic locus. AB - AIMS: Characterization of the aggregation-promoting factor (APF) of the human intestinal isolate Lactobacillus crispatus M247 and its homologous nonaggregating mutant Mu5. METHODS AND RESULTS: Western blot analysis revealed that the supernatant of both M247 and Mu5 contains a 28-kDa protein which cross reacts with the antiserum produced against the APF of Lact. gasseri 4B2. The apf genes of M247 and Mu5 strains were identical and were shown to be 672 nucleotides in length and encoding a protein of 223 amino acids with a predicted molecular weight of 24.0 kDa. CONCLUSION: Our results shows that the lost of aggregation in Mu5 is not related to a defect in secretion of the APF protein or a mutation in the apf gene. SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest that the mutation in Mu5 may be contained in another molecule involved in aggregation such as a possible receptor for APF. PMID- 15357725 TI - Expression of vip1/vip2 genes in Escherichia coli and Bacillus thuringiensis and the analysis of their signal peptides. AB - AIMS: To determine the expression time courses and high expression level of Vip2A(c) and Vip1A(c) in Bacillus thuringiensis, and survey their insecticidal toxicity and insecticidal spectrum. METHODS AND RESULTS: A kind of new vegetative insecticidal toxin genes encoded by a single operon from B. thuringiensis had been cloned and sequenced. The individual genes, 5-terminus truncated genes and the operon were respectively expressed in Escherichia coli. Only N-terminus deleted Vip2A(c) and Vip1A(c) proteins could be purified by Ni-NTA agarose, while others were processed and their N-terminal signal peptides were cleaved. The individual genes and the operon were also expressed in B. thuringiensis. Both proteins were mostly secreted into the cell supernatants. The expression level of Vip1A(c) was influenced because of the interruption of vip2A(c) gene on the operon. Bioassays showed that neither separate protein nor both performed any toxicity against tested lepidopteran and coleopteran insects. CONCLUSIONS: Vip2A(c) and Vip1A(c) have similar secretion mechanism in E. coli and B. thuringiensis. Vip1A(c) remained its high expression level only when being expressed with vip2A(c) gene as an operon in B. thuringiensis. SIGNIFICANCE AND IMPACT OF THE STUDY: Expression of vip2A(c) and vip1A(c) genes in E. coli and B. thuringiensis were investigated. This would help to make clear the secretion mechanism of VIP proteins and study the function of ADP-ribosyltransferase Vip2. PMID- 15357726 TI - Identification of extracellular enzyme producing alkalophilic bacilli from Izmir province by 16S-ITS rDNA RFLP. AB - AIMS: To screen industrially important extracellular enzymes from the newly isolated alkalophilic bacilli and to characterize them by phenotypic and 16S internal transcribed spacer (ITS) rDNA restriction pattern analysis. METHODS AND RESULTS: Three different environmental samples, soil, leather and horse faeces, were collected within the province of Izmir. Isolates grown on Horikoshi-I medium for 24 h at 37 degrees C were screened for extracellular enzyme activity by using eight different substrates: birchwood xylan, carboxymethylcellulose, casein, citrus pectin, polygalacturonic acid, soluble starch, and Tween 20 and 80. In total, 115 extracellular enzyme-producing bacilli were obtained. Casein was hydrolysed by 78%, soluble starch by 67%, citrus pectin by 63%, polygalacturonic acid by 62%, Tween 20 by 34%, birchwood xylan by 16%, Tween 80 by 12%, and carboxymethylcellulose by 3% of the isolates. The isolates were differentiated into 19 distinct homology groups by the 16S-ITS rDNA restriction pattern analysis. CONCLUSIONS: Eight different extracellular enzyme activities were determined in 115 endospore forming bacilli. The largest 16S-ITS rDNA homology group (HT1) included 36% of the isolates, 98% of which degraded casein, polygalacturonic acid, pectin and starch. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report on the characterization of the industrial enzyme producing alkalophilic bacilli by 16S-ITS rDNA restriction fragment length polymorphism (RFLP). Restriction profiles of 64% of the isolates were found to be different from those of five reference strains used. PMID- 15357727 TI - Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum. AB - AIM: Understanding the molecular response to stress tolerance of wine Lactobacillus plantarum. METHODS AND RESULTS: Two genes codifying for heat shock proteins were cloned from wine L. plantarum. The coding regions of the two heat shock genes are 420 and 444 nucleotides long, and started with an ATG codon suggesting that they were translated. The protein sequences deduced from the isolated genes have a molecular mass of 18.483 and 19.282 kDa, respectively, and were therefore named hsp18.5 and hsp19.3. The expression of small heat shock genes was analysed by RT-PCR analysis. Moreover, the 5' and 3' noncoding regions were cloned and sequenced. CONCLUSIONS: The expression of the heat shock genes was strongly induced by heat, cold and ethanol stress. Analysis of the 5' and 3' flanking regions of hsp18.5 and hsp19.3 genes, revealed the presence of an inverted repeat sequence (TTAGCACTC-N(9)-GAGTGCTAA) homologue to the CIRCE elements found to the upstream regulatory region of heat shock operons, and an inverted sequence that could form a stem and loop structure that it is likely to function as a transcriptional terminator. Based on their structures, the genes were classified as belonging to Class I of heat shock genes according to the B. subtilis nomenclature of heat response genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Small heat shock genes isolated from wine L. plantarum might have a role in preventing damage by cold stress. PMID- 15357728 TI - Optimization of extracellular endoxylanase, endoglucanase and peroxidase production by Streptomyces sp. F2621 isolated in Turkey. AB - AIMS: To determine the effect of environmental conditions on the production of extracellular lignocellulose-degrading enzymes by Streptomyces sp. F2621 and to assess the potential use of these enzymes in the hydrolysis of lignocellulose material. METHODS AND RESULTS: The production of extracellular lignocellulose degrading enzymes, endoxylanase, endoglucanase and peroxidase during the growth of Streptomyces sp. F2621 in basal salts-yeast extract medium containing different carbon sources and the effect of a number of environmental parameters (e.g. carbon sources and concentrations, pH and temperature) were investigated. The highest endoxylanase (22.41 U ml(-1)) and peroxidase (0.58 U ml(-1)) activities were obtained after 2-4 days of incubation at 30 degrees C in a basal salts medium containing 0.4% (w/v) oat spelt xylan and 0.6% (w/v) yeast extract, corresponding to C : N ratio of 6 : 1. Cell-free extracellular enzyme preparations from the strain were capable of releasing both sugar and aromatic compounds during incubation with eucalyptus paper pulp, straw and xylan. Overall, 9.3% hydrolysis of xylan occurred after 24-h incubation. However the rates of hydrolysis of paper pulp and straw were approximately twofold less than xylan hydrolysis, although the total percentage hydrolysis of available substrate (24.5% and 16.3%, respectively) was greater than xylan hydrolysis. CONCLUSIONS: The high levels of enzyme production achieved under batch cultivation conditions, coupled with no significant production of endoglucanase during the growth phase of organism and the release of both sugar and aromatic compounds from paper pulp and straw signify the suitability for these enzymes for industrial applications such as pulp and paper production. SIGNIFICANCE AND IMPACT OF THE STUDY: The results highlight the environmental conditions for the production of extracellular lignocellulose-degrading enzymes by Streptomyces sp. F2621 and suggest the use of streptomycetes and/or their enzymes in industrial processes. PMID- 15357729 TI - New PCR-based methods for yeast identification. AB - AIMS: To characterize reference yeast strains and identify indigenous strains isolated from wine fermentations by PCR methods. METHODS AND RESULTS: We compared several PCR techniques for yeast identification. We used oligonucleotide primers that are complementary to (i) intron splice sites, (ii) REP and (iii) ERIC elements to produce PCR fingerprints that display specific patterns between the different yeast species. These three techniques were used to characterize 41 reference yeast strains belonging to 15 different species and to identify 40 indigenous strains isolated from grape must and wine fermentations. Species specific banding patterns were obtained with the three PCR-techniques with different degrees of intraspecific differentiation depending on the method. By comparing the PCR fingerprints of unknown isolates with those produced by reference strains, we identified yeast strains isolated from an industrial wine fermentation. CONCLUSIONS: All three PCR techniques are rapid, reliable and simple methods of yeast identification. As far as we know, this is the first time that the primers designed for amplifying repetitive elements in bacteria have been successfully used in yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: Industry needs rapid, reliable and simple methods of yeast identification. The proposed PCR techniques will allow to achieve this objective. PMID- 15357730 TI - Morphological and phenotypical characterization of Bacillus sporothermodurans. AB - AIMS: Enumeration of resistant bacteria in ultra-high temperature (UHT) treated milk; morphological characterization and phenotyping of resistant strains by traditional and nontraditional methods and their identification by molecular biology. METHODS AND RESULTS: Modified standard plate count agar (PCA) and modified brain-heart infusion (BHI) agar were used for colony counts. Physiological culture traits were determined as suggested by Bergey's Manual of Systematic Bacteriology or in modified J-broth or in modified BHI agar. Scanning electron microscope (SEM) was used for microscopic examination. Strain identification was carried out by polymerase chain reaction (PCR). A total of 125 (62.81% of 199) samples were positive and the bacterial load was higher than 10(5) CFU ml(-1) in 46 samples (28.80% of 125). The 16S rRNA sequence of bacterial cultures obtained from UHT-treated milk was similar to that of Bacillus sporothermodurans M215 type strain((T)) and different biotypes were found by analysis of colony appearance, cell morphology and physiological traits. CONCLUSIONS: Bacillus sporothermodurans was the predominant sporigenous micro organisms in UHT milk. SIGNIFICANCE AND IMPACT OF THE STUDY: BHI agar is more suitable than PCA for quality control of milk after UHT treatment. Modified J broth medium is useful to determine selected physiological traits of B. sporothermodurans. The strains characterized and identified as B. sporothermodurans were significantly different compared with the type strain. PMID- 15357731 TI - Identification of extracellular enzyme producing thermophilic bacilli from Balcova (Agamemnon) geothermal site by ITS rDNA RFLP. AB - AIMS: Molecular characterization of extracellular enzyme producing thermophilic bacilli from Balcova geothermal site. METHODS AND RESULTS: Three types of geothermal samples were collected: mud, re-injection water, and samples from uncontrolled hydrothermal vents. Isolates grown at 55 degrees C in culture media prepared in sterilized re-injection water, were screened for extracellular enzyme activity by using eight different substrates: casein, carboxymethylcellulose, pectin, polygalacturonic acid (PGA), soluble starch, Tween 20 and 80, and xylan. In total, 109 thermoaerophilic isolates were selected. All of the isolates could hydrolyse Tween 20 (100%) but not Tween 80. Soluble starch was hydrolysed by 96%, casein by 55%, xylan and carboxymethylcellulose by 9%, and pectin and PGA by 2% of the isolates. The isolates were grouped into 14 different homology groups by the restriction pattern analysis of 16S-internal transcribed spacer (ITS) rDNA RFLP. Each of the RFLP groups was also studied by 16S rRNA gene partial sequence analysis. Plasmid DNA profiles revealed that 15 of the isolated strains contained small plasmid DNA molecules ranging in size from 12 000 to 35 000 bp. CONCLUSIONS: Combined analysis of 16S-ITS rDNA RFLP and 16S rRNA gene partial sequence results indicated the presence of novel or existing species of Anoxybacillus (nine species) and Geobacillus (three species). SIGNIFICANCE AND IMPACT OF THE STUDY: In this study 16S-ITS rDNA RFLP was applied for the first time to differentiate thermophilic bacilli. It was also the first study on thermophilic bacilli of Balcova geothermal site. PMID- 15357732 TI - The effect of Propionibacterium acidipropionici, with or without Lactobacillus plantarum, on the fermentation and aerobic stability of wheat, sorghum and maize silages. AB - AIMS: To determine the effect of Propionibacterium acidipropionici, alone or in combination with Lactobacillus plantarum, on the fermentation and aerobic stability of wheat, sorghum and maize silages. METHODS AND RESULTS: The inoculants were applied at 1.0 x 10(6) CFU g(-1). Silages with no additives served as control. Fresh forages were sampled prior to ensiling. Three jars per treatment were sampled on days 2, 4, 8, 16 and 60 after ensiling, for chemical and microbiological analysis. At the end of the ensiling period, the silages were subjected to an aerobic stability test. The P. acidipropionici-inoculated silages had significantly higher levels of acetic and propionic acid than the L. plantarum or P. acidipropionici + L. plantarum-inoculated silages (P < 0.05). Therefore, yeast activity was impaired in the P. acidipropionici-inoculated silages. As a result, P. acidipropionici decreased CO(2) production and improved aerobic stability of wheat, sorghum and maize silages. However, the combination of P. acidipropionici + L. plantarum did not improve aerobic stability of the silages. CONCLUSIONS: The P. acidipropionici was very effective in protecting the wheat, sorghum and maize silages exposed to air under laboratory conditions, probably because the acidic environment under ensiling conditions is favourable for this micro-organism. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of P. acidipropionici, as a silage inoculant can improve the aerobic stability of silages by inhibition of yeast activity. PMID- 15357733 TI - Identification of denitrifying rhizobacteria from bentgrass and bermudagrass golf greens. AB - AIMS: As high rates of nitrogen fertilization are used in turfgrass management, there is a great potential for nitrogen loss. Research on identification of denitrifiers in turfgrass has been limited. Therefore, the aim was to identify denitrifier species and genes from turfgrass roots. METHODS AND RESULTS: Rhizobacteria were isolated from roots of bentgrass and bermudagrass in sand based United States Golf Association (USGA) golf greens and used for denitrification biochemical analysis. Seventeen per cent (34 isolates) were identified as denitrifiers, 47% were classified as nitrate-reducers and 36% were nondenitrifiers. Identification of species of the denitrifiers was performed by chromatography fatty acid methyl ester (GC-FAME) and16S rDNA analyses. Bacillus and Pseudomonas were the major turfgrass denitrifiers. The two methods showed a 60% agreement at the genus level. Nitrite reductase genes nirK and nirS were detected in 74 and 15% of the denitrifiers, respectively, but not in nondenitrifiers. The nosZ gene encoding nitrous oxide reductase was detected in all the denitrifiers, but also in some nondenitrifiers. CONCLUSIONS: To our knowledge, this is the first report for identification of denitrifiers and denitrification-related genes associated with turfgrass roots. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide valuable data for future denitrification studies that seek to improve turfgrass nitrogen management for maximum efficiency. PMID- 15357734 TI - Treatment with oxidizing agents damages the inner membrane of spores of Bacillus subtilis and sensitizes spores to subsequent stress. AB - AIMS: To determine if treatment of Bacillus subtilis spores with a variety of oxidizing agents causes damage to the spore's inner membrane. METHODS AND RESULTS: Spores of B. subtilis were killed 80-99% with wet heat or a variety of oxidizing agents, including betadine, chlorine dioxide, cumene hydroperoxide, hydrogen peroxide, Oxone, ozone, sodium hypochlorite and t-butylhydroperoxide, and the agents neutralized and/or removed. Survivors of spores pretreated with oxidizing agents exhibited increased sensitivity to killing by a normally minimal lethal heat treatment, while spores pretreated with wet heat did not. In addition, spores treated with wet heat or the oxidizing agents, except sodium hypochlorite, were more sensitive to high NaCl in plating media than were untreated spores. The core region of spores treated with at least two oxidizing agents was also penetrated much more readily by methylamine than was the core of untreated spores, and spores treated with oxidizing agents but not wet heat germinated faster with dodecylamine than did untreated spores. Spores of strains with very different levels of unsaturated fatty acids in their inner membrane exhibited essentially identical resistance to oxidizing agents. CONCLUSIONS: Treatment of spores with oxidizing agents has been suggested to cause damage to the spore's inner membrane, a membrane whose integrity is essential for spore viability. The sensitization of spores to killing by heat and to high salt after pretreatment with oxidizing agents is consistent with and supports this suggestion. Presumably mild pretreatment with oxidizing agents causes some damage to the spore's inner membrane. While this damage may not be lethal under normal conditions, the damaged inner membrane may be less able to maintain its integrity, when dormant spores are exposed to high temperature or when germinated spores are faced with osmotic stress. Triggering of spore germination by dodecylamine likely involves action by this agent on the spore's inner membrane allowing release of the spore core's depot of dipicolinic acid. Presumably dodecylamine more readily alters the permeability of a damaged inner membrane and thus more readily triggers germination of spores pretreated with oxidizing agents. Damage to the inner spore membrane by oxidizing agents is also consistent with the more rapid penetration of methylamine into the core of treated spores, as the inner membrane is likely the crucial permeability barrier to methylamine entry into the spore core. As spores of strains with very different levels of unsaturated fatty acids in their inner membrane exhibited essentially identical resistance to oxidizing agents, it is not through oxidation of unsaturated fatty acids that oxidizing agents kill and/or damage spores. Perhaps these agents work by causing oxidative damage to key proteins in the spore's inner membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: The more rapid heat killing and germination with dodecylamine, the greater permeability of the spore core and the osmotic stress sensitivity in outgrowth of spores pretreated with oxidizing agents is consistent with such agents causing damage to the spore's inner membrane, even if this damage is not lethal under normal conditions. It may be possible to take advantage of this phenomenon to devise improved, less costly regimens for spore inactivation. PMID- 15357735 TI - A novel method for isolation of Campylobacter spp. from environmental samples, involving sample processing, and blood- and antibiotic-free medium. AB - AIMS: To develop a method that involves sample processing, and blood- and antibiotic-free medium for isolation and enumeration of Campylobacter spp. from environmental samples. METHODS AND RESULTS: The sample processing (preT) was standardized to minimize the population of competing bacteria. A blood- and antibiotic-free differential, Kapadnis-Baseri medium (KB medium) was formulated and tested for isolation of Campylobacter spp. in comparison with CAT medium. PreT-KB method was evaluated in comparison with the conventional viable count method and with the conventional most probable number (C. MPN) method for enumeration of Campylobcater from environmental samples. The results indicated that sample processing significantly reduced population of competing bacteria. The KB medium selected Gram-negative bacteria and differentiated Campylobacter from lactose-fermenting competing bacteria. The population of Campylobacter detected by preT-KB method was similar to that by conventional viable count method. While, the population of Campylobacter spp. determined by preT-KB method was higher than that by C. MPN method. In addition, the preT-KB method detected antibiotic sensitive campylobacters. CONCLUSION: The preT minimizes population of competing bacteria and the KB medium selects Gram-negative bacteria and differentiates Campylobacter from them. Therefore, Campylobacter can be isolated from environmental samples without using antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: The preT-KB method is simple and facilitates isolation of antibiotic sensitive and enumeration of Campylobacter in the environmental samples. Therefore, the new method will be useful for isolation and enumeration of Campylobacter from water, food and sewage samples. Besides, it would also detect antibiotic-sensitive campylobacters, which are not detected by conventional viable count and MPN methods. PMID- 15357736 TI - The incidence of Shiga toxin-producing Escherichia coli in cattle with mastitis in Brazil. AB - AIMS: To determine the prevalence and molecular characteristics of Shiga toxin producing Escherichia coli (STEC) isolates from bovine mastitic milk in Brazil. METHODS AND RESULTS: A total of 2144 milk samples from dairy cattle showing mastitis were screened for the presence of E. coli. A total of 182 E. coli isolates were selected and examined. All were subjected to dot blot analysis using the CVD419 probe for the detection of the enterohaemolysin (hly) gene, and to a multiplex PCR for the detection of stx1, stx2 and eaeA genes. STEC were isolated from 22 (12.08%) milk samples. All the STEC isolates were tested for sensibility to 10 antimicrobials; the resistances most commonly observed were to cephalothin (86.3%), tetracycline (63.6%) and doxycycline (63.6%). CONCLUSION: STEC isolates were found in bovine mastitic milk in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: STEC isolates from mastitic milk were potentially pathogenic for human in that they belonged to serogroups associated with diarrhoea and haemolytic-uraemic syndrome, some of them were stx2, eaeA and hly positive. PMID- 15357737 TI - Repression of reserve lipid turnover in Cunninghamella echinulata and Mortierella isabellina cultivated in multiple-limited media. AB - AIMS: To study patterns of reserve lipid biosynthesis and turnover (degradation) in two oleaginous Zygomycetes, namely Cunninghamella echinulata and Mortierella isabellina under various growth conditions. Fatty acid composition of the reserve lipid of both strains was also studied in all growth steps. METHODS AND RESULTS: Cunninghamella echinulata and Mortierella isabellina were grown in carbon-excess batch cultures. In the investigated strains, accumulation of reserve lipid occurred only when the activity of both NAD(+)-isocitrate dehydrogenase (ICDH) and NADP(+)-ICDH were not detectable in the cell-free extract. Specifically, in C. echinulata, NAD(+)-ICDH activity was detected even after depletion of ammonium nitrogen in the medium, resulting in a delay of the initiation of lipid accumulation period. On the contrary, in M. isabellina, lipid accumulation occurred simultaneously with ammonium nitrogen exhaustion in the growth medium, as the activity of both NAD(+)- and NADP(+)-ICDH were not detectable after nitrogen depletion. In C. echinulata reserve lipid was not degraded after glucose had been exhausted. Supplementations of the medium with Fe(3+), yeast extract or Mg(2+) induced, however, reserve lipid breakdown and formation of lipid-free material. In M. isabellina after glucose exhaustion, notable lipid degradation occurred, accompanied by a significant lipid-free material biosynthesis. Nevertheless, in multiple-limited media, in which Mg(2+) or yeast extract, besides carbon and nitrogen, were limiting nutrients, reserve lipid breakdown was repressed. In both strains, the quantity of gamma-linolenic acid (GLA) in the reserve lipids [varying between 9 and 16% (w/w) in C. echinulata and 1.5-4.5% (w/w) in M. isabellina] was proportional to lipid-free biomass. CONCLUSIONS: Lipid accumulation period in Zygomycetes is initiated by the attenuation of ICDH activity in the mycelium while the regulation of ICDH from ammonium nitrogen is strain specific. While a single nitrogen limitation was enough to induce lipid accumulation, however, multiple limitations were needed in order to repress lipid turnover in oleaginous Zygomycetes. As for GLA, its biosynthesis in the mycelium seemed proportional to lipid-free biomass synthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: Several nutrients are indispensable for functioning the mechanisms involved in the mobilization of reserve lipid in oleaginous moulds. Therefore, reserve lipid turnover in oleaginous moulds could be repressed in multiple limited media. PMID- 15357738 TI - Reduction of alpha-galactooligosaccharides in soyamilk by Lactobacillus fermentum CRL 722: in vitro and in vivo evaluation of fermented soyamilk. AB - AIMS: Consumption of soya-derived products has been hampered by the presence of alpha-galactooligosaccharides (alpha-GOS) because mammals lack pancreatic alpha galactosidase (alpha-Gal) which is necessary for their hydrolysis. These sugars thus reach the large intestine causing gastrointestinal disorders in sensitive individuals. The use of lactic acid bacteria (LAB) expressing alpha-Gal is a promising solution for the degradation of alpha-GOS in soyamilk. METHODS AND RESULTS: The capacity of the LAB Lactobacillus fermentum CRL 722 to properly degrade alpha-GOS was studied in vitro using controlled fermentation conditions and in vivo using a rat model. Lactobacillus fermentum CRL 722 was able to grow on commercial soyamilk and completely eliminated stachyose and raffinose during fermentation because of its high alpha-Gal activity. Rats fed soyamilk fermented by this LAB had smaller caecums compared with rats fed unfermented soyamilk. CONCLUSIONS: Soyamilk fermentation by Lact. fermentum CRL 722 results in the reduction of alpha-GOS concentrations in soyamilk, thus eliminating possible undesirable physiological effects normally associated with its consumption. SIGNIFICANCE AND IMPACT OF THE STUDY: Fermentation with Lact. fermentum CRL 722 could prevent gastrointestinal disorders in sensitive individuals normally associated with the consumption of soya-based products. This LAB could thus be used in the elaboration of novel fermented vegetable products which better suit the digestive capacities of consumers. PMID- 15357739 TI - Radiation recall dermatitis. PMID- 15357740 TI - Cutaneous manifestations associated with antiphospholipid antibodies. AB - BACKGROUND: Primary Antiphospholipid Antibody Syndrome (PAAS) is characterized by detection of antiphospholipid antibodies associated with venous or arterial thrombosis and/or miscarriages by patients with no other associated disease such as systemic lupus erythematosus (SLE). Primary Antiphospholipid Antibody Syndrome has many clinical manifestations of which dermatological ones are probably the most common. The purpose of this study was to determine the frequency of each cutaneous lesion, describing clinical features essential for diagnosis, in patients with Antiphospholipid Antibody Syndrome (AAS) attending Walter Cantidio University Hospital. METHODS: Sixty patients with clinical findings suggestive of AAS were screened, and submitted for clinical and laboratory evaluations including lupus anticoagulant (KCT), anticardiolipin antibodies (IgG and IgM: ELISA), routine laboratory tests and screening tests for possible associated conditions. RESULTS: Twenty-five cases of primary and 14 cases of secondary AAS were diagnosed by clinical and laboratory evidences. Persistent elevated antiphospholipid antibodies without history of thromboembolic events or miscarriages were demonstrated in 21 patients. Forty percent of the patients with AAS had a cutaneous feature as the major complaint. These were dermographism (15), acrocyanosis (13), urticaria (9), diffuse alopecia (9), livedo reticularis (seven), Raynaud's phenomenon (three), purpura (two), ulcers and necrosis (four), nodules (four), pterygium ungueum (one) and subungual hemorrhage (one). CONCLUSIONS: Dermatological complaints are very frequent in patients with AAS and may be the first clue to the syndrome. Therefore a careful history and detailed physical examination are essential to diagnose AAS. All dermatologists should investigate the possibility of AAS when facing cutaneous findings related to venous or arterial thrombosis or microthrombosis. PMID- 15357741 TI - Awareness of, knowledge of and attitudes to nonmelanoma skin cancer (NMSC) and actinic keratosis (AK) among physicians. AB - BACKGROUND: Nonmelanoma skin cancer (NMSC) is the most common type of cancer that affects the Caucasian population. Approximately 80% of NMSCs are basal cell carcinoma (BCC) and 20% are squamous cell carcinoma (SCC). Actinic keratosis (AK) is a precancerous lesion that may develop into SCC. METHODS: A market research survey was conducted in which dermatologists and primary care physicians (PCPs) were randomly selected from seven countries (USA, Australia, UK, Italy, France, Germany and Spain). Their knowledge of nonmelanoma skin cancer and their current clinical practice were assessed. RESULTS: In total, 2100 physicians took part in the survey. They had practised medicine for between 1 and 30 years and saw at least 30 patients in a typical week. The majority of dermatologists (97%) were familiar with BCC and AK, and treated each condition with a minimum of referrals. PCPs were more familiar with BCC (90%) than with AK (74%). Of the PCPs that were aware of BCC, only 31% treated the condition, and of those aware of AK, 40% treated the condition. Surgery was the most common choice of treatment for BCC. The most popular treatment choice for AK lesions was cryotherapy. Eighty to 100% of physicians reported that they discussed skin cancer prevention with their patients. A much lower number of physicians (ranging from 5 to 37%) provided educational material to patients. Overall, PCPs in the two countries that have a high incidence of NMSC (USA and Australia) were more familiar with BCC and AK and more likely to treat each condition than PCPs in Europe. All physicians rated BCC as a more serious condition than AK. Facial lesions were considered more serious than lesions on the head or trunk for both conditions. CONCLUSIONS: As the burden of disease and the number of patients seeking treatment for NMSC increase, dermatologists are well placed to lead educational initiatives for PCPs and provide educational material for patients. This would increase awareness of AK and BCC and could improve early diagnosis. PMID- 15357742 TI - Epidemiology of Kaposi's sarcoma in patients with acquired immunodeficiency syndrome in Sao Paulo, Brazil. AB - BACKGROUND: Kaposi's sarcoma (KS) is the most frequent neoplasm in patients with acquired immunodeficiency syndrome (AIDS). Although many studies on KS epidemiology have been performed in other countries, few have been carried out in Brazil despite the high incidence of AIDS. METHODS: One hundred and seven KS patients seen in Sao Paulo, Brazil, between August 1995 and November 1998 were studied. The patients were followed for 1 year with assessment of the immunologic status, improvement of the lesions, treatment, and causes of death at the end of this period. RESULTS: KS occurred mainly in men (94.4%) with a mean age of 37 years, and 25.2% of these patients were found to be human immunodeficiency virus (HIV) seropositive through KS. HIV was acquired mainly through homosexual contact. The patients presented an average of 15.9 KS lesions at the first visit and the mean duration of KS lesions before the first visit was 15.5 months. The clinical presentation was predominantly papules and plaques, and 33.6% presented with mucosal and/or visceral disease. KS affected mainly the lower limbs. The mean time since the diagnosis of HIV infection was 42.4 months. The CD4+ cell count was lower than 200 cell/mm(3) in 60.8% of patients, but patients with a complete response showed an improvement in immune status after 1 year. Patients who did not show progression received a protease inhibitor as part of highly active antiretroviral therapy (HAART). Patients treated exclusively with HAART presented a complete response (61.6%), partial response (23%) or progression (15.4%) of KS. CONCLUSIONS: An improvement in immune status and the use of HAART were the most important prognostic features. PMID- 15357743 TI - Cutaneous hyalohyphomycosis caused by Paecilomyces lilacinus: report of three cases and review of the literature. AB - Cutaneous infection with Paecilomyces lilacinus is encountered worldwide, with most infections occurring either iatrogenically or in immunocompromised hosts. We report three cases of cutaneous hyalohyphomycosis caused by P. lilacinus, one of which occurred in an immunocompetent individual. In addition, we review the 17 cases previously reported in the literature. Although this infection can be difficult to treat, most cases are not fatal. Most cases responded well to systemic azole antifungal agents, either alone or in association with surgical excision of the lesion. Paecilomyces species are saprophytic fungi found in soil and decaying organic matter. Infection in humans is rare, but when it occurs treatment is often difficult. We report three cases of cutaneous hyalohyphomycosis caused by P. lilacinus and review the 17 previously reported cases in which the clinical history and response to therapy were described. PMID- 15357744 TI - Epidemiology of childhood psoriasis: a study of 419 patients from northern India. AB - BACKGROUND: We undertook this study in order to determine the pattern and prevalence of childhood psoriasis in northern India and to highlight the differences and similarities with previous studies. MATERIALS AND METHODS: In this retrospective epidemiologic study, the data from 419 children (less than 14 years of age) with psoriasis registered at the Psoriasis Clinic between January 1990 and December 2002 were included. RESULTS: The 419 children registered at the Psoriasis Clinic constituted 0.3% of the dermatology outpatients and 12.5% of the total psoriasis patients seen over a period of 13 years in the department. There were 219 (52.2%) boys and 200 (47.7%) girls, with a male to female ratio of 1.09 : 1. The age of onset ranged from 4 days to 14 years. The mean age of onset was 8.1 +/- 2.1 years in boys and 9.3 +/- 2.3 years in girls. The peak age of onset in boys was in the 6-10-year age group, whereas the majority of girls showed an onset of psoriasis between the ages of 10 and 14 years. A positive family history was present in only 19 (4.5%) patients. The extensors of the legs were the most common initial site affected [105 (25%) cases], followed by the scalp [87 (20.7%)]. Classical plaque psoriasis was the most frequent clinical presentation [254 (60.6%) patients], followed by plantar psoriasis [54 (12.8%)]. Nail involvement was observed in 130 (31%) cases. All types of nail changes described in psoriasis were seen in these patients. Pitting was the most common nail change, followed by ridging and discoloration. Five children (1.1%) (three girls and two boys) had psoriatic arthropathy. Precipitating factors that brought about the onset of the disease or were associated with exacerbation could be recalled in only 28 (6.6%) patients. Koebnerization was observed in 27.9% of patients. Pruritus was the most frequent symptom, reported by 365 (87.1%) children. Twenty seven (6.4%) children had other concurrent mucocutaneous diseases (vitiligo, pityriasis alba, alopecia areata, ichthyosis vulgaris, halo nevus, aphthous stomatitis, urticaria, pityriasis versicolor, nummular eczema, salmon patch, and verrucous epidermal nevus). Eighteen children had systemic disorders, including seizures, bronchial asthma, mitral regurgitation, scleroderma, Down's syndrome, high arched palate, cholelithiasis, anterior mongoloid slant, and prognathism; however, these conditions were possibly chance findings only and no correlation with the age of onset or severity of the disease was found. CONCLUSIONS: Our findings differ from those of previous studies in showing a delayed onset, equal sex distribution, less frequent facial involvement, uncommon guttate lesions, more frequent involvement of the soles, and a less frequent history of familial occurrence. PMID- 15357745 TI - Itraconazole in the treatment of New World mucocutaneous leishmaniasis. AB - BACKGROUND: A well-tolerated oral drug is required for the treatment of mucocutaneous leishmaniasis (MCL). Current parenteral treatment regimens with pentavalent antimonials are associated with marked toxicity and significant rates of relapse. AIM: To evaluate the efficacy and tolerability of high-dose itraconazole for the treatment of MCL. METHODS: An uncontrolled treatment study was performed in 13 Ecuadorian patients with MCL. Each patient received a daily dosage of 400 mg of itraconazole for a minimum of 3 months. RESULTS: All 13 subjects responded to itraconazole during the first month of treatment, but by 12 months after treatment the complete resolution of MCL lesions was observed in only three (23%) subjects. No adverse effects of treatment were reported. Response to treatment was associated with a short evolution of the disease and mild to moderate disease severity. CONCLUSION: Prolonged and high-dose treatment regimens with itraconazole are not effective for the treatment of the majority of patients with MCL. PMID- 15357746 TI - Cupping: East meets West. PMID- 15357747 TI - Zosteriform metastatic melanoma. PMID- 15357748 TI - Cutaneous sarcoid-like granulomas with alveolar hemorrhage and c-ANCA PR-3. PMID- 15357749 TI - Case of disseminated Langerhans' cell histiocytosis presenting with sclerosing cholangitis. PMID- 15357750 TI - Intertriginous xanthomas: a marker of homozygous type IIa hyperlipoproteinemia. PMID- 15357751 TI - Combined basal cell carcinoma and Langerhans cell histiocytosis of the scrotum in a patient with occupational exposure to coal tar and dust. PMID- 15357752 TI - Penile self-injections: an unusual act. PMID- 15357753 TI - Pemphigus vulgaris with involvement of the cervix treated using thalidomide therapy. PMID- 15357754 TI - Prolidase deficiency: case reports of two Argentinian brothers. PMID- 15357755 TI - A prospective study of the use of cryosurgery for the treatment of actinic keratoses. AB - BACKGROUND: Actinic keratoses are the most common actinic lesions on Caucasian skin. Cryosurgery with liquid nitrogen is commonly used to treat actinic keratoses, but there have been few studies examining the true rate of cure in everyday dermatologic practice. AIM: To determine prospectively the true efficacy of cryosurgery as a treatment for actinic keratoses in everyday dermatologic practice. METHODS: A prospective, multicentered study (a subsidiary study of a photodynamic therapy trial) was performed. Patients with untreated actinic keratoses greater than 5 mm in diameter on the face and scalp were recruited. Eligible lesions received a single freeze-thaw cycle with liquid nitrogen given via a spray device and were reviewed 3 months thereafter. Each center used their preferred freeze time. The only treatment criterion was complete freezing of actinic keratoses and a 1-mm rim of normal skin. Treated lesions were assessed as complete response or noncomplete response. The influence of the duration of freeze, cosmetic outcomes, and adverse events were examined. RESULTS: Ninety adult patients from the community with 421 eligible actinic keratoses were recruited. The overall individual complete response rate was 67.2%[SEM = +/-3.5%; 95% confidence interval (CI) = 60.4-74.1%]. Complete response was 39% for freeze times of less than 5 s, 69% for freeze times greater than 5 s, and 83% for freeze times greater than 20 s. Cosmetic outcomes were good to excellent in 94% of complete response lesions. The main adverse events were pain, stinging, and burning during treatment, and hypopigmentation after healing. CONCLUSIONS: Cryosurgery is an effective treatment for actinic keratoses. The true complete response rate is significantly lower than that previously reported. The freeze duration influences successful treatment. Adverse events are mild and well tolerated. PMID- 15357756 TI - Alpha-1-antitrypsin deficiency associated with panniculitis treated with plasma exchange therapy. AB - BACKGROUND: Alpha-1-antitrypsin is the principal serum protease inhibitor. In addition to the well-recognized association with early-onset emphysema and cirrhosis, alpha-1-antitrypsin deficiency may be associated with panniculitis. The treatment of this type of panniculitis presents a significant challenge. Previous attempts using immunosuppressive, anti-inflammatory, and cytotoxic drugs have shown variable results. AIM: To report a case of alpha-1-antitrypsin deficiency-associated panniculitis treated with plasma exchange therapy. METHODS: A 23-year-old patient developed painful red nodules on her thighs and buttocks with spontaneous ulceration and discharge of oily fluid. A skin biopsy specimen showed septal and lobular panniculitis. The serum alpha-1-antitrypsin level was 22 mg/dL. She was treated with plasma exchange therapy. RESULTS: Treatment of this patient with plasma exchange therapy led to the control of the cutaneous lesions. CONCLUSIONS: Plasma exchange therapy represents an alternative treatment which restores serum and tissue alpha-1-antitrypsin levels. This method is proposed because of its clinical benefits and greater availability. PMID- 15357757 TI - Alpha-1-antitrypsin deficiency-associated panniculitis. PMID- 15357758 TI - Co-occurrence does not imply association. PMID- 15357759 TI - Excessive drowsiness with low-dose methotrexate. PMID- 15357760 TI - Further insight into the pathomechanism of acne by considering the 5-alpha reductase inhibitory effect of linoleic acid. PMID- 15357765 TI - Liver biopsy in patients with inherited disorders of coagulation and chronic hepatitis C. AB - Liver biopsy plays a pivotal role in the management of patients with a variety of liver diseases, including chronic hepatitis C virus. The major risk of the procedure is the potential for significant haemorrhagic complications. Although the data are limited, the procedure does not appear to pose excessive risk to the patient with inherited disorders of coagulation, provided that adequate haemostasis can be achieved prior to the liver biopsy. This requires close coordination of care between the hepatologist and the haematologist. Indications for liver biopsy should be the same in patients with haemophilia as in other populations. PMID- 15357766 TI - Radiation synovectomy by 188Re-Sulfide in haemophilic synovitis. AB - Radiation synovectomy is a safe and effective treatment for chronic haemophilic synovitis causing recurrent haemarthroses. This study reports results of 29 188Re sulfide radiation synovectomies in knee joint with a follow-up 18 months. Using the absorbed dose factor, three groups of radioactivity dose (555 MBq, 687 MBq and 917 MBq) were used according to the synovium thickness that measured by MRI. Excellent and good results (haemarthrosis reduction from 75 to 100%) were obtained in 71% of cases at 18 months. Comparison of the results, no significant difference was found among the radioactivity dose groups. Radiation was well contained within the knee. There were no observed or identified complications including the detectable harm to the articular cartilage, as shown by MRI. PMID- 15357767 TI - Clinical evaluation of an advanced category antihaemophilic factor prepared using a plasma/albumin-free method: pharmacokinetics, efficacy, and safety in previously treated patients with haemophilia A. AB - The efficacy and safety of an advanced category recombinant antihaemophilic factor produced by a plasma- and albumin-free method (rAHF-PFM) was studied in 111 previously treated subjects with haemophilia A. The study comprised a randomized, double-blinded, crossover pharmacokinetic comparison of rAHF-PFM and RECOMBINATE rAHF (R-FVIII); prophylaxis (three to four times per week with 25-40 IU kg(-1) rAHF-PFM) for at least 75 exposure days; and treatment of episodic haemorrhagic events. Median age was 18 years, 96% of subjects had baseline factor VIII <1%, and 108 received study drug. Bioequivalence, based on area under the plasma concentration vs. time curve and adjusted in vivo recovery, was demonstrated for rAHF-PFM and R-FVIII. Mean (+/-SD) half-life for rAHF-PFM was 12.0 +/- 4.3 h. Among 510 bleeding events, 473 (93%) were managed with one or two infusions of rAHF-PFM and 439 (86%) had efficacy ratings of excellent or good. Subjects who were less adherent to the prophylactic regimen had a higher bleeding rate (9.9 episodes subject(-1) year(-1)) than subjects who were more adherent (4.4 episodes subject(-1) year(-1); P < 0.03). One subject developed a low titre, non-persistent inhibitor (2.0 BU) after 26 exposure days. These data demonstrate that rAHF-PFM is bioequivalent to R-FVIII, and suggest that rAHF-PFM is efficacious and safe, without increased immunogenicity, for the treatment of haemophilia A. PMID- 15357768 TI - Clinical efficacy, safety and pharmacokinetic properties of the factor VIII concentrate Haemoctin SDH in previously treated patients with severe haemophilia A. AB - Clinical efficacy, safety and pharmacokinetic properties of the high-purity double-virus inactivated plasma-derived factor VIII concentrate Haemoctin SDH (pdFVIII) were evaluated in three prospective open-label uncontrolled studies in previously treated patients (PTPs) with severe haemophilia A. The pharmacokinetic properties assessed at baseline and after 3 months of treatment are in accurate accordance with published data and remain unchanged over time (study A, n = 12). Mean terminal elimination half-life was 11.8 and 11.9 h, mean incremental recovery (IU dL(-1)/IU kg(-1)) was 2.3 and 2.0, respectively. Long-term efficacy and safety, in particular the potential immunogenicity, were investigated in a total of 53 PTPs (studies A and B) treated prophylactically and on-demand, as required. PdFVIII has shown to be effective in preventing and controlling bleeding episodes; 23.5% of patients were free of bleeding events. A total of 177 haemorrhages occurred with 74.0% resolving after a single infusion, 87.6% within two infusions. 98.3% of responses reported on haemorrhages were rated as 'excellent' or 'good'. Moreover, 'excellent' haemostatic efficacy has been demonstrated in 10 surgical procedures including general and severe orthopaedic interventions (study C). No complication occurred in any surgery. Few adverse events were reported, one patient developed a high-titre FVIII inhibitor without clinical relevance. In all three studies, over 6 million units were administered in nearly 4300 infusions, approximately 94% units or infusions were given for prophylaxis and only 6% for treatment on-demand. In conclusion, pdFVIII has shown to be effective, safe and well tolerated in long-term prophylaxis and treatment on-demand as well as after minor and major surgical procedures. PMID- 15357769 TI - Increased breakthrough bleeding during prophylaxis with B-domain deleted factor VIII--a robust meta-analytic finding. AB - Meta-analyses of observational studies have become increasingly common to support evidence-based clinical decisions. We analyzed currently available clinical studies of full-length factor VIII (FL-FVIII) vs. B-domain deleted recombinant factor VIII (BDD-rFVIII) using a random effects model to investigate possible differences in clinical efficacy in patients treated during prophylaxis. Some studies reported breakthrough bleeding incidence as mean annual total bleeds, whereas others reported median bleeds. In accord with the consensus recommendations by the Meta-analysis of Observational Studies in Epidemiology (MOOSE) group, all available studies where included. For analysis, data were combined by converting median to mean annual total bleeds using a conversion factor of 2.6, based on clinical data previously compiled by the Universal Data Collection Program of the U.S. Centers for Disease Control and Prevention. To evaluate the sensitivity of our model upon the choice of conversion factor, we re estimated incidence rate ratios for breakthrough bleeding over a wide range of conversion factors from 1.4-2.6. Even at the lowest extreme conversion factor of 1.4, bleeding incidence was statistically higher in patients treated with BDD rFVIII compared with FL-FVIII. We also examined the impact of reported patient age on our multivariate model. Exposure to BDD-rFVIII remained an independent predictor of bleeding, regardless of patient age at start or mean age during prophylaxis. These analyses further support the robustness of our meta-analysis and its conclusions. PMID- 15357770 TI - Continuous infusion of B-domain deleted recombinant factor VIII (ReFacto) in patients with haemophilia A undergoing surgery: clinical experience. AB - This retrospective, open-label, non-comparative study evaluated continuous infusion of recombinant factor VIII (ReFacto), B-domain deleted recombinant FVIII (BDDrFVIII), in patients with haemophilia A undergoing surgery and requiring >5 consecutive days of treatment. Sixteen patients from eight centres underwent a total of 20 procedures. Haemostatic outcome was assessed as 'excellent' or 'good' in 75% of procedures, and target FVIII:C levels were maintained throughout the continuous infusion period. The reported volume of blood loss during surgery was also within the normal range for non-haemophilic patients for the type of surgery performed. Red blood cell transfusions were required to balance excessive blood loss during BDDrFVIII continuous infusion in eight (40%) procedures (seven patients), five with bleeding or requiring volume replacement and three to treat anaemia secondary to blood loss. Non-serious adverse events considered by investigators as possibly or probably related to BDDrFVIII continuous infusion were infrequent (n = 5) considering the duration of treatment (n =239 cumulative days of continuous infusion), and all of these were mild-to-moderate in severity. No thromboembolic complications were reported except for one case of thrombophlebitis occurring at the infusion site. Only two patients (four events) experienced serious adverse bleeding; BDDrFVIII was otherwise well-tolerated. These data show that continuous infusion of BDDrFVIII provides reliable haemostasis and is an effective and well-tolerated regimen for patients with haemophilia A undergoing surgery. PMID- 15357771 TI - Relationships between factor VIII:Ag and factor VIII in recombinant and plasma derived factor VIII concentrates. AB - A variety of plasma-derived (pd) and recombinant (r) factor VIII (FVIII) concentrates are used to prevent and treat bleeding in severe hemophilia A patients. A significant side effect of FVIII replacement is the development of FVIII neutralizing antibodies (inhibitors) in up to 30% of patients receiving FVIII concentrates. The FVIII protein content (FVIII:Ag) per unit of FVIII:C in FVIII concentrates, and how effectively the FVIII:Ag in FVIII concentrates binds to von Willebrand factor (VWF) may provide information relevant for the survival of FVIII:C in vivo and for estimating the risk for inhibitor development. The FVIII:Ag content of nine r-FVIII and nine pd-FVIII concentrates were quantified in this study using two enzyme-linked immunosorbent assay (ELISA) platforms. The two ELISA platforms were based on the use of a monoclonal anti-(FVIII light chain)-IgG and polyclonal anti-FVIII antibodies as capture antibodies and both ELISAs were equally able to detect > or =0.005 IU of FVIII:Ag. Measured in international units, the r-FVIII concentrates contained significantly higher FVIII:Ag per unit of FVIII:C than the pd-FVIII concentrates. The VWF-binding profiles of the r-FVIII and pd-FVIII concentrates were also determined by gel filtration chromatography. Unlike the plasma-derived products, the r-FVIII concentrates invariably contained a fraction of FVIII:Ag molecules (approximately 20%) which was unable to associate with VWF. Given that VWF regulates both factor VIII proteolysis and survival of FVIII:Ag in vivo, the fraction of FVIII:Ag unable to bind to VWF may have a reduced survival and be more susceptible to proteolytic degradation in vivo. The extent to which the fractions of FVIII:Ag in concentrates able and unable to bind to VWF contribute to inhibitor development in severe FVIII-deficient patients is unknown. PMID- 15357772 TI - Factor XIII deficiency in south-east Iran. PMID- 15357773 TI - Type 2N von Willebrand disease mutations in Brazilian individuals. PMID- 15357774 TI - Endometrial ablation for von Willebrand disease-related menorrhagia--experience with seven cases. AB - BACKGROUND: Endometrial ablation has recently gained popularity as a treatment of menorrhagia in the general population. In the von Willebrand disease (VWD) patient, intuitively, it would appear that the failure rate would be higher because of the underlying hypocoaguability increasing the likelihood for re bleeding. In a consecutive series of seven patients, we assessed the efficacy and safety of endometrial ablation in VWD-related menorrhagia. PATIENTS AND METHODS: We performed a retrospective analysis using chart review and a 21-item questionnaire administered to seven (six type 1, one type 2A) women who underwent endometrial ablation between the years 1997 and 2001. Parameters assessed included operative complications, the development of abdominal pains, recurrence of menstrual bleeding post-ablation and the change in the pre-/post-ablation quality of life (QOL). Three patients underwent endomyometrial resection and one each underwent rollerball, thermal, electrocautery and balloon ablation. All patients were pre-treated with i.v. desmopressin (DDAVP) except the 2A patient who received Humate P. Mean age of the patient was 41 +/- 6 years and follow-up was for 45 months (range 31-73) post-ablation. RESULTS: No significant perioperative bleeding complications were observed in any of the patients. All patients initially responded (two amenorrhoea, four hypomenorrhoea, one moderate improvement). In all patients, QOL assessed by 10 parameters improved significantly following the ablation procedure, regardless of the specific technique used. However, at the end of follow-up, only one patient remained amenorrheic, one was hypomenorrheic, one had moderate improvement and four patients experienced recurrence of menorrhagia, three eventually requiring a hysterectomy at a median of 11 months post-ablation. CONCLUSION: Endometrial ablation appears to be a safe procedure that improves the QOL in patients with VWD-related menorrhagia. However, its long-term efficacy appears to be lower in VWD patients when compared with women with menorrhagia without VWD. PMID- 15357775 TI - Clinical manifestations and management of labor and delivery in women with factor IX deficiency. AB - Haemophilia is uncommon in females and little is known about the clinical manifestations and postpartum management of women with this disorder. Clinical characteristics of postpartum bleeding were evaluated in women with factor IX deficiency (FIX:C < 0.20 U mL(-1)), including two with haemophilia B and three carriers of haemophilia B, undergoing labour and delivery. Data were collected prospectively during routine outpatient comprehensive haemophilia care at the haemophilia Center of Western Pennsylvania and during inpatient management. Four of five women experienced postpartum bleeding, during six of 16 deliveries: the median haemoglobin was 10.7 g% and two required blood transfusion. Postpartum bleeding was significantly more common among those receiving fewer than 4 days of FIX replacement: six of 13 (46.1%) receiving fewer three or fewer days bled vs. none of three (0%) receiving six or more days treatment [P < 01 (Wilcoxon)]. Postpartum bleeding was not related to the route of delivery (P = 0.525), vaginal vs. Caesarean, nor the FIX level (P = 0.371; FIX > 0.05 U mL(-1) vs. < or =0.05 U mL(-1)). Compared with females with von Willebrand disease or FXI deficiency, females with FIX deficiency were more likely to experience postpartum bleeding (P = 0.008) and anaemia (P = 0.045); and they were less likely to experience menorrhagia (P = 0.065), but the latter did not reach significance. Postpartum bleeding is common in women with haemophilia B or carriers of haemophilia B, and treatment with factor replacement for at least 4 days of postpartum may prevent bleeding following delivery in such women. PMID- 15357776 TI - Inhibitor development in patients receiving recombinant factor VIII (Recombinate rAHF/Bioclate): a prospective pharmacovigilance study. AB - Clinical trials to date have not been adequately powered to assess comparatively infrequent events such as inhibitor development in previously treated patients (PTPs). Comprehensive large-scale pharmacovigilance studies can be useful for this purpose. We prospectively collected inhibitor development reports worldwide among recipients of Recombinate rAHF recombinant factor VIII (rFVIII), also formerly distributed under the product name Bioclate, for the entire postlicensure period from 1993 through 2002. To determine level of exposure to rFVIII we also compiled the Recombinate rAHF/Bioclate International Units (IU) distributed annually. To estimate inhibitor incidence separately for previously untreated or minimally treated patients (PUPs) with 1-50 exposure days and PTPs with >50 exposure days, we used haemophilia A incidence and prevalence data and pooled mean annual rFVIII consumption per PUP and PTP from international multicentre prospective clinical trials. Documented inhibitor cases totalled 89, and the total quantity of Recombinate rAHF/Bioclate rFVIII distributed was 6.48 x10(9) IU. No lot association or other clustering of inhibitor events was evident in PTPs. The incidence of all reported inhibitors, expressed as a percentage of patients treated, was 11.9% (CI: 5.05-28.0%) for PUPs when compared with 0.123% (CI: 0.030-0.512%) for PTPs. The rates for high-titre inhibitors (>5 BU) only were 5.96% (CI: 3.00-11.8%) for PUPs and 0.0554% (CI: 0.0113-0.271%) for PTPs. Thus, incidence rates for both all inhibitors and high-titre inhibitors in PTPs were 1% of the corresponding rates in PUPs. Data from prospective PUP clinical trials involving intensive active monitoring suggest that true inhibitor incidence may be approximately twice that estimated in this pharmacovigilance study. Nevertheless, inhibitor development in PTPs receiving Recombinate rAHF/Bioclate is infrequent. PMID- 15357777 TI - Treatment patterns and cost-of-illness of severe haemophilia in patients with inhibitors in Germany. AB - To evaluate current treatment patterns and resource utilization as well as related cost in the management of severe haemophilia patients with inhibitors in Germany, a cost-of-illness study was conducted. Generally, data were generated by structured literature search. Missing data were collected by expert interviews. All data were validated by a panel of German experts in haemophilia care. In Germany, immune tolerance therapy (ITT) is first-line therapy in inhibitor management for children in the initial year after inhibitor development, particularly for high responders (HR). In adult HR patients ITT is applied but to a remarkably lower extent than in children. To treat bleeding episodes, factor VIII (FVIII) is first-line therapy in low responders (LR). For paediatric HR patients, bleeds are mainly treated with recombinant FVIIa (rFVIIa). In adult HR patients, activated prothrombin complex concentrate (aPCC) and rFVIIa are more equally distributed as treatment options. Treatment costs were calculated for paediatric patients (15 kg) and adult patients (75 kg) from third party payers' perspective. Cost for ITT ranges from Euro 70,290 (2 months; LR) to Euro 3 812,400 (24 months; with aPCC; HR) in a paediatric patient. For an adult patient ITT cost ranges from Euro 287,500 (6 months; LR) to Euro 17,253,000 (36 months; HR). For on average 12.5 acute bleeds, average annual treatment costs amount to Euro 77,000 for a child and Euro 354,000 for an adult. Assessing the results it has been taken into consideration that ITT can last longer and annual number of bleeds can be extremely higher than on average 12.5 episodes. This indicates more health care resource consumption in some patients. PMID- 15357778 TI - HLA class II genotype and factor VIII inhibitors in mild haemophilia A patients with an Arg593 to Cys mutation. AB - We evaluated inhibitor formation in a group of patients with mild haemophilia A caused by an Arg593 to Cys mutation. A remarkably high cumulative inhibitor incidence of 14% over 22 years was observed. Three of 49 patients developed transient, low-titre inhibitors, which remained below 2.0 BU mL(-1). Four patients with an Arg593 to Cys mutation developed high-titre inhibitors (>5.0 BU mL(-1)). Three of these patients have been described previously. In this study, we characterized inhibitory antibodies in a fourth patient with high-titre inhibitors. Epitope mapping studies revealed that antibodies were predominantly directed to the A2 domain of factor VIII. We addressed the role of human leucocyte antigen (HLA) class II alleles in inhibitor development in patients with an Arg593 to Cys mutation by HLA genotyping. In the group of inhibitor patients raised frequencies of HLA-DRB1*01 and HLA-DQB1*05 were observed that did not reached statistical significance. Our data suggest that inhibitor development in mild haemophilia A patients with an Arg593 to Cys mutation is not linked to HLA class II profile. PMID- 15357779 TI - Costs of on-demand and prophylactic treatment for severe haemophilia in Norway and Sweden. AB - The expected annual cost (in the year 2000 prices) for a 30-year-old patient with average individual and treatment characteristics for on-demand EUR 51,832 (95% CI: 44,324-59,341) and for prophylaxis EUR 146,118 (95% CI: 129,965-162,271), was obtained from panel-data analysis of an 11-year retrospective panel of 156 patients with severe haemophilia in Norway and Sweden. Costs included haemophilia related treatment costs within the health-care sector (factor concentrate, doctors' visits, diagnostic procedures, hospitalisation, invasive procedures, etc.) and cost for haemophilia-related resource use in other sectors (lost production, use of special equipment, adaptation of workplace and domicile, etc). Although costs of lost production, reconstructive surgery and hospitalisation were higher for on-demand, they did not balance out the higher costs of factor concentrate consumption in prophylaxis. The cut-off risk of premature death, where on-demand and prophylaxis would have been equally costly, was 3.7 percentage units higher for on-demand than for prophylaxis. Such a great risk difference has not been reported elsewhere to our knowledge. Estimated cost elasticities indicated that annual costs of prophylaxis would increase by approximately the same proportion as a potential increase in the price of factor concentrate and decrease less than proportionately with a reduction in prescribed dose kg(-1). For on-demand, the annual costs would increase by approximately the same proportion as an increase in the prescribed dose kg(-1). PMID- 15357780 TI - Willingness to pay for on-demand and prophylactic treatment for severe haemophilia in Sweden. AB - The objective of the present paper was to provide an estimate of the benefits of on-demand and prophylaxis treatment strategies for severe haemophilia in monetary terms. Using the contingent-valuation method, which simulates a missing market by asking people about their willingness to pay (WTP), we asked a representative sample (n = 609) of the Swedish population if they would be willing to pay a specific amount (bid) so that patients with severe haemophilia could receive on demand treatment and another bid for prophylactic treatment. Different respondents were offered different bids and the bid vector ranged from 71 Euro cents to EUR 130. The order of the bid questions was randomized so that half of the respondents were asked first about their WTP for on-demand treatment, and then about their WTP for prophylaxis, while the order was reversed for the other half of the respondents. The mean estimated WTP (year 2002) was EUR 39 (95% CI 31 47) for on-demand and EUR 65 (95% CI 55-73) for prophylaxis. Our sensitivity analysis showed that the ranking of the two treatment alternatives was robust in that the WTP was greater for prophylaxis in all possible subsets. The point estimates of WTP varied somewhat in subsets defined by individual characteristics, but confidence intervals always overlapped that of the main results. The WTP for on-demand and prophylaxis exceeded the calculated cost of treatment per taxpayer of providing on-demand and prophylactic treatment, respectively, based on our previous results. PMID- 15357781 TI - Survival analysis of patients with haemophilia at the International Haemophilia Training Centre, Bangkok, Thailand. AB - Haemophiliac treatment in less developed countries is limited to locally prepared fresh frozen plasma, cryoprecipitate, cryo-removed plasma and lyophilized products as replacement therapy. Factor concentrate is seldom used because of the high price. The present study reports the survival analysis of 164 patients comprising 138 haemophilia A and 26 haemophilia B cases from 134 families registered at the International Haemophilia Training Centre-Bangkok, Faculty of Medicine, Ramathibodi Hospital, Mahidol University from 1971 to 2000. The duration of follow-up ranged from 1 to 27 years and 8 months with a median of 9 years and 1 month. They received treatment on demand of bleedings, and 85 patients received additional home treatment for early bleedings. The proportion of death was 15.2%. The Kaplan-Meier survival curves revealed the median (95% CI) survival time of patients with severe and moderate degrees of 35 years and 6 months (21.4-49.7), and 38 years and 1 month (28.8-47.3), respectively. To compare the progressive achievement of haemophilia care services, the treatment period was divided into three decades: 1971-1980, 1981-1990 and 1991-2000. The patients with unaided proper walking increased from 62.8% in the first decade to 84.5% in the third decade. However, one-third of the patients developed one to four chronic haemarthrosis determined by clinical evaluation, especially patients with severe degree. Moreover, the estimated probability of a survival time beyond 13 years of age among patients with severe degree increased from 0.85 in the first decade to 0.94 and 1 in the second and third decades, respectively. Thus, established haemophilia care, even with limited resources, has significantly decreased the risk of death and increased the survival time among patients with haemophilia. PMID- 15357782 TI - Novel missense mutation in the coagulation factor IX catalytic domain associated with severe haemophilia B--Factor IXDelhi. AB - Factor IX is a vitamin K-dependent serine protease, which exists as a zymogen in the blood. On activation to factor IXa, by factor XIa or tissue factor-factor VIIa complex, it forms tenase complex with factor VIIIa, in the presence of Ca2+. This tenase complex enzymatically converts factor X to factor Xa, thereby bringing about the coagulation cascade. Mutations in factor IX gene have been shown to cause haemophilia B, which is inherited as an X-linked recessive disorder. Herein we report a novel missense mutation at the nucleotide position 30829-T > A in the exon 8 of factor IX gene. This transversion leads to the substitution of histidine 236 to glutamine. This resulting abnormal protein has been named factor IXDelhi. Molecular modelling was performed to predict the molecular pathology of this mutation. We predict that this change in the catalytic domain may affect the surface loop that accommodates Ca2+, thereby leading to severe bleeding disorder. PMID- 15357783 TI - Informativeness of linkage analysis for genetic diagnosis of haemophilia A in India. AB - The objective of this study was to assess the frequency of factor VIII (FVIII) gene intron 1 and intron 22 inversions and the informativeness of polymorphic markers for the genetic diagnosis of patients with haemophilia A (HA). Fifty unrelated patients with HA were first assessed for the intron 1 and intron 22 inversion mutations. Inversion-negative families were then screened for the bi allelic intragenic markers--intron 7 G-->A polymorphism, HindIII site in intron 19 and XbaI site in intron 22 and the multiallelic dinucleotide CA repeat alleles in introns 13 and 22. The extragenic, multiallelic VNTR DXS52 (st14) was also analysed. Intron 22 inversion mutation was found in 38% (n = 19) of all patients and 46% of those with severe HA. Intron 1 inversion was found in one (2%) patient. Of the 30 inversion-negative families, XbaI site polymorphism was the single most informative marker (70%, n = 21/30) followed by HindIII (60%, n = 18/30), intron 13 CA repeats (56.66%, n = 17/30), intron 22 CA repeats (50%, n = 15/30), DXS52 VNTR (23.33%, n = 7/30) and intron 7 G-->A polymorphism (6.66%, n = 2/30). The combined use of these markers was informative in 92% (n = 46/50) of HA families. Based on the informativeness of these markers a comprehensive algorithm has been proposed for genetic diagnosis of HA in India. PMID- 15357784 TI - The impact of unaware carriership on the clinical presentation of haemophilia. AB - This study was conducted to evaluate the effect of an unaware carriership on the delay in diagnosis of haemophilia and the resulting effect of this delay on morbidity. Information on 73 haemophilia patients (<18 years) and their mothers was gathered from data of patients' medical records and completed by interviews with the parent(s). Although a positive family history was present in 52 gravidae, 16 of them (31%) were not aware of their carrier status at moment of delivery. Fifteen of these 16 unaware carriers, were carriers of a non-severe form of haemophilia. In mothers who were unaware of carriership for haemophilia instrumental delivery occurred more frequently than in mothers who knew they were carriers. This is disquieting since instrumental delivery poses a significant risk (relative risk: 17.8, 95% CI: 4.0-78.4) for intra- or extracranial bleedings in newborn haemophiliacs in comparison to spontaneous deliveries or caesarean sections. In 83% of the patients with a positive family history, diagnosis was established before the first bleeding episode. Patients diagnosed by bleedings presented more often with iatrogenic bleedings (38%) then patients who were diagnosed because of a positive family history (9%) (P < 0.05). In comparison to previous studies, more patients had a positive family history and in more haemophiliacs with a positive family history diagnosis was established before the first bleeding episode. Although this reflects the yield of improved diagnostic methods, further reduction of iatrogenic bleeding is possible by intensifying counselling of mild haemophilia carriers. PMID- 15357785 TI - Functional consequences of haemophilia in adults: the development of the Haemophilia Activities List. AB - Several instruments can be used to evaluate the functional status of patients with haemophilia, but none of these instruments is specific for haemophilia. We developed a haemophilia-specific self-assessment questionnaire to evaluate and monitor a patient's perceived functional health status: the Haemophilia Activities List (HAL). In three separate but interlinked substudies, the questionnaire was constructed and tested for face, expert, and convergent validity, as well as internal consistency and patient-evaluated relevance. Items for the questionnaire were collected by interviewing 162 patients, using the McMaster-Toronto Arthritis Patient Preference Disability Questionnaire (MACTAR). The items were combined to generate the first version of the questionnaire [HAL(1)]. This version was evaluated and commented on by two focus groups (patients and caregivers), and then the questionnaire was adapted on the basis of these comments, forming the final version, HAL(2). This version was then validated in a pilot study with 50 consecutive patients using the Dutch Arthritis Impact Measurements Scales 2 (Dutch-AIMS2) and the Impact on Participation and Autonomy (IPA) questionnaires. The HAL(2) showed good convergent validity (Pearson correlation 0.80-0.91; P < 0.01), and the internal consistency was good for six of the eight domains (Cronbach's alpha 0.83-0.95). Patients considered the content of the HAL to be more relevant to their situation than the content of the other questionnaires (P < 0.01). Three major factors (upper extremity function, lower extremity function, key activities/major problem activities) were identified by factor analysis. The questionnaire seems to be a useful tool to identify problematic activities as part of the functional health status of patients with haemophilia. The construct validity, test-retest reliability, and responsiveness of the HAL will be established in the future. PMID- 15357786 TI - Evaluation of the impact of information about treatment-related risks in patients receiving blood-derived or recombinant medications. AB - The aims of the study were to evaluate the impact of a written information about treatment related risks in patient receiving blood derived or recombinant medications. Haemophiliac patients and patients with constitutional or acquired immune deficiencies are concerned by this treatment and these information. Our objectives are to evaluate the efficacy of the written information, the knowledge of the patients about these medications and the psychological, emotional impact if these information. The study is based on questionnaires which specified how the patient treat bleeding episodes, their knowledge about viral safety of blood products, the patient's perception of his or her health status and relationship with the physician. Psychological and emotional status are evaluated with the Hospital Anxiety and Depression Scale. The results show the difficulty to inform patients: if the information generate only limited anxiety in patients with haemophilia or immune deficiencies, we observe that the delivery of a written information got a mediocre effect on overall knowledge. We think that this information must be appropriate for patients and be communicated orally within the patient-physician relationship. PMID- 15357787 TI - Asking fathers: a study of psychosocial adaptation. AB - Although few contemporary studies specifically address paternal adaptation, the theme of paternal estrangement from medical care and from family relationships is pervasive in the psychosocial literature on haemophilia. This estrangement has been shown to have a negative effect on fathers' psychological well-being, marital relationships and the adaptive outcome of their sons who have haemophilia. The goals of this study were to provide contemporary data on the psychosocial adaptation of fathers of boys with haemophilia and to examine specific variables that might influence their adjustment. Eighty-three eligible fathers returned a survey instrument that collected demographic and medical information, as well as scores on self-measures of adaptation in marital and parenting roles. Statistically significant direct correlations (P < 0.01) were found between fathers' scores on the Marital Adjustment Test and the Parenting Sense of Competence subscales (parenting efficacy and satisfaction). Variables specific to rearing a son with haemophilia that negatively affected fathers' marital adjustment scores included: feeling left out of medical decision making by their wives or partners, worry about their sons' having limited activity, and the presence of a secondary diagnosis in the affected child. Scores on the parenting efficacy subscale of the PSOC were statistically significantly reduced (i.e. fathers felt less effective in the parenting role) in men who 'rarely' or 'never' infused their sons (42/80, 53%). Variables that negatively affected scores on the parenting satisfaction subscale included frustrating interactions with medical staff and concern about their sons' potential to contract an infection or secondary diagnosis. This paper presents a model to examine the interrelationships among the data and discusses the clinical implications. PMID- 15357788 TI - Yttrium synoviorthesis of the elbow joints in persons with haemophilia. AB - Yttrium has been used as the material of choice for synoviorthesis in our centre since 1984. An audit was carried out to determine the long-term results of this modality for treating elbow joints. An unacceptably high percentage of patients lost range-of-movements in the articulation treated. Good results were achieved with respect to a reduction in the bleeding incidences. It is suggested that another isotope be utilized and that other centres carry out an audit and publish their data. PMID- 15357789 TI - The rare coagulation disorders--review with guidelines for management from the United Kingdom Haemophilia Centre Doctors' Organisation. AB - The rare coagulation disorders are heritable abnormalities of haemostasis that may present significant difficulties in diagnosis and management. This review summarizes the current literature for disorders of fibrinogen, and deficiencies of prothrombin, factor V, FV + VIII, FVII, FX, the combined vitamin K-dependent factors, FXI and FXIII. Based on both collective clinical experience and the literature, guidelines for management of bleeding complications are suggested with specific advice for surgery, spontaneous bleeding, management of pregnancy and the neonate. We have chosen to include a section on Ehlers-Danlos Syndrome because haematologists may be consulted about bleeding manifestations in such patients. PMID- 15357790 TI - Consensus recommendations for use of central venous access devices in haemophilia. AB - Venous access is essential for delivery of haemophilia factor concentrate. Wherever possible, peripheral veins remain the route of choice, and the use of central venous access devices (CVADs) should be limited to cases of clear need in patients with caregivers able to exercise diligence in CVAD care and should continue no longer than necessary. CVADs are of recognized value for repeated administration of coagulation factors in haemophilia, particularly for prophylaxis and immune tolerance therapy and in young children. Evidence to guide best practices has been fragmentary, and standardized methods for CVAD usage have yet to be established. We have developed management recommendations based upon available published evidence as well as extensive clinical experience. These recommendations address patient and CVAD selection; CVAD placement, care and removal; caregiver/patient guidance; and complications, including infection and thrombosis. In the absence of inhibitors, ports are recommended, primarily because of fewer associated infections than with external catheters. For patients with inhibitors, ports also appear to be associated with fewer infections. Infection is the most frequent complication, and recommendations to prevent and treat infections are supported by extensive clinical data and experience. Strict adherence to handwashing and aseptic technique are essential elements of catheter care. Evidence-based data regarding the detection and treatment of CVAD-related thrombotic complications are limited. Caregiver education is an integral part of CVAD use and the procedural practices of users should be regularly re-assessed. These recommendations provide a basis for sound current CVAD practice and are expected to undergo further refinements as new evidence is compiled and clinical experience is gained. PMID- 15357791 TI - Shoulder hemiarthroplasty to manage haemophilic arthropathy: two case studies. AB - Bleeds within the shoulder joint can lead to significant joint destruction and may have associated pain, decreased range of movement (ROM), and impaired function. Conservative management should involve prompt administration of Factor VIII and physiotherapy to address all surrounding structures so as to minimize further damage. If conservative management fails to relieve severe, unremitting shoulder pain in the presence of underlying arthropathy, then arthroplasty may be considered. Outcomes of arthroplasties performed for osteoarthritis and rheumatoid arthritis appear favourable. Few articles, however, have addressed shoulder arthroplasty to manage haemophilic arthropathy, and no reports have documented the rehabilitation process. Three hemiarthroplasties were performed at the Royal Brisbane and Women's Hospital on two men with haemophilia. There were no surgical or postoperative complications. Rehabilitation included intensive physiotherapy. The results in each case revealed a decrease in pain, and an increase in ROM and function postoperatively. These findings suggest that hemiarthroplasty with postoperative physiotherapy may be a feasible option to manage severe, chronic and progressing shoulder pain as a result of haemophilic arthropathy of the shoulder. PMID- 15357792 TI - Successful use of ReFacto continuous infusion in two paediatric patients with severe haemophilia A undergoing orthopaedic surgery. AB - In this report we describe the successful use of B-domain-deleted recombinant factor VIII (ReFacto) administered by continuous infusion during orthopaedic procedures in two children with severe haemophilia A. Both patients underwent ankle synovectomy and in patient 2, a medial patello-femoral ligament repair was performed in the same operative session. Patient 2 developed septic arthritis A in his knee joint and arthroscopic joint irrigation and debridement was performed 2 weeks after the initial procedure. Surgical cover was initiated with a bolus dose of ReFacto 50 IU kg(-1) followed by continuous infusion at 3.3-4.8 IU kg(-1) h(-1) which was maintained for up to 9 days postoperatively. Patient 2 received an additional bolus dose of 15 IU kg(-1) during the infusion period. All procedures were performed without haemostatic complications and long-term orthopaedic outcomes were good in both patients. PMID- 15357793 TI - Acquired inhibitor of factor V: first report in China and literature review. AB - We reported a 51-year-old man who developed a severe haematuria with a prolonged prothrombin time (PT) and activated partial thromboplastin time (aPTT). Laboratory evaluation revealed the presence of specific factor V inhibitors. No underlying diseases and associated conditions have been identified. The patient improved after immunosuppressive therapy. However, the FV level remained low, indicating incomplete inhibitor eradication. The clinical course of our patient was consistent with the observations by others that FV inhibitor can be persistent for months or years. PMID- 15357794 TI - Severe factor X deficiency in pregnancy: case report and review of the literature. AB - Isolate factor X deficiency is an extremely rare clotting factor disorder inherited in autosomal recessive fashion and pregnancy in a homozygous patient is frequently complicated by recurrent miscarriage, uterine bleeding and premature labour. Eleven pregnancies in seven patients affected by FX deficiency have been reported in the literature. Two additional pregnancies have been reported in a FX variant (FX Friuli). We present a new case of successful at term pregnancy in a homozygous patient. PMID- 15357795 TI - Haemostatic management of intraoral bleeding in patients with congenital deficiency of alpha2-plasmin inhibitor or plasminogen activator inhibitor-1. AB - Haemostatic management of intraoral bleeding was investigated in patients with congenital alpha2-plasmin inhibitor (alpha2-PI) deficiency or congenital plasminogen activator inhibitor- 1 (PAI-1) deficiency. When extracting teeth from patients with congenital alpha2-PI deficiency, we advocate that 7.5-10 mg kg(-1) of tranexamic acid be administered orally every 6 h, starting 3 h before surgery and continuing for about 7 days. For the treatment of continuous bleeding, such as post-extraction bleeding, 20 mg kg(-1) of tranexamic acid should be administered intravenously, and after achieving local haemostasis 7.5 mg kg(-1) of tranexamic acid should be administered orally every 6 h for several days. In addition, when treating haematoma caused by labial or gingival laceration or buccal or mandibular contusion, haemostasis should be achieved by administering 7.5-10 mg kg(-1) of tranexamic acid every 6 h. Tranexamic acid can also be used for haemostatic management of intraoral bleeding in patients with congenital PAI 1 deficiency, but is less effective when compared with use in patients with congenital alpha2-PI deficiency. Continuous infusion of 1.5 mg kg(-1) h(-1) of tranexamic acid is necessary for impacted tooth extraction requiring gingival incision or removal of local bone. PMID- 15357796 TI - Factors which may influence coping with disease in haemophilia patients. PMID- 15357797 TI - Comparative effectiveness of plasma-derived and recombinant FVIII concentrates in the on-demand and prophylactic treatment of patients with haemophilia A. PMID- 15357798 TI - Letter to the editor regarding "strategic alternative responses to risks of terrorism (11 october, 2003)". PMID- 15357799 TI - Risk assessment for invasive species. AB - Although estimates vary, there is a broad agreement that invasive species impose major costs on the U.S. economy, as well as posing risks to nonmarket environmental goods and services and to public health. The domestic effort to manage risks associated with invasive species is coordinated by the National Invasive Species Council (NISC), which is charged with developing a science-based process to evaluate risks associated with the introduction and spread of invasive species. Various international agreements have also elevated invasive species issues onto the international policy agenda. The World Trade Organization (WTO) Sanitary and Phytosanitary (SPS) Agreement establishes rights and obligations to adhere to the discipline of scientific risk assessment to ensure that SPS measures are applied only to the extent required to protect human, animal, and plant health, and do not constitute arbitrary or unjustifiable technical barriers to trade. Currently, however, the field of risk assessment for invasive species is in its infancy. Therefore, there is a pressing need to formulate scientifically sound methods and approaches in this emerging field, while acknowledging that the demand for situation-specific empirical evidence is likely to persistently outstrip supply. To begin addressing this need, the Society for Risk Analysis Ecological Risk Assessment Specialty Group and the Ecological Society of America Theoretical Ecology Section convened a joint workshop to provide independent scientific input into the formulation of methods and processes for risk assessment of invasive species to ensure that the analytic processes used domestically and internationally will be firmly rooted in sound scientific principles. PMID- 15357800 TI - Allee effects and the risk of biological invasion. AB - The Allee effect is a nonlinear phenomenon exhibited in the population dynamics of sparse populations in which the per capita population growth rate increases with increasing population density. In sufficiently sparse populations, the Allee effect may lead to extinction and is known to generate a threshold in the probability of establishment when presented as a function of introduced population size or density. As introduced populations are generally small, Allee effects are probably common in biological invasions and their consideration is necessary for accurately assessing the risk of invasion by many species, including all sexually reproducing species. Bythotrephes longimanus, an invasive, freshwater, cladoceran zooplankter from Europe, is one such species. Here, I review a previously published model of the Allee effect for continuously sexually reproducing species. Then, I develop a new model for seasonally parthenogenetic species such as Bythotrephes, and thereby demonstrate the potential consequences of Allee effects. This result underscores the importance of considering nonlinear phenomena, including thresholds, when conducting risk analysis for biological introductions. PMID- 15357801 TI - Assessing the risk of invasive spread in fragmented landscapes. AB - Little theoretical work has investigated how landscape structure affects invasive spread, even though broad-scale disturbances caused by habitat loss and fragmentation are believed to facilitate the spread of exotic species. Neutral landscape models (NLMs), derived from percolation theory in the field of landscape ecology, provide a tool for assessing the risk of invasive spread in fragmented landscapes. A percolation-based analysis of the potential for invasive spread in fragmented landscapes predicts that invasive spread may be enormously enhanced beyond some threshold level of habitat loss, which depends upon the species' dispersal abilities and the degree of habitat fragmentation. Assuming that invasive species spread primarily through disturbed areas of the landscape, poor dispersers may spread better in landscapes in which disturbances are concentrated in space, whereas good dispersers are predicted to spread better in landscapes where disturbances are small and dispersed (i.e., fragmented landscape). Assessing the risk of invasive spread in fragmented landscapes ultimately requires understanding the relative effects of landscape structure on processes that contribute to invasive spread--dispersal (successful colonization) and demography (successful establishment). Colonization success is predicted to be highest when >20% of the landscape has been disturbed, particularly if disturbances are large or aggregated in space, because propagules are more likely to encounter sites suitable for colonization and establishment. However, landscape pattern becomes less important for predicting colonization success if species are capable of occasional long-distance dispersal events. Invasive species are also more likely to persist and achieve positive population growth rates (successful establishment) in landscapes with clumped disturbance patterns, which can then function as population sources that produce immigrants that invade other landscapes. Finally, the invasibility of communities may be greatest in landscapes with a concentrated pattern of disturbance, especially below some critical threshold of biodiversity. Below the critical biodiversity threshold, the introduction of a single species can trigger a cascade of extinctions among indigenous species. The application of NLMs may thus offer new insights and opportunities for the management and restoration of landscapes so as to slow the spread of invasive species. PMID- 15357802 TI - Projecting rates of spread for invasive species. AB - All else being equal, the faster an invading species spreads, the more dangerous its invasion. The projection of spread rate therefore ought to be a central part of the determination of invasion risk. Originally formulated in the 1970s to describe the spatial spread of advantageous alleles, integrodifference equation (IDE) models have since been co-opted by population biologists to describe the spread of populations. More recently, they have been modified to include population structure and environmental variability. We review how IDE models are formulated, how they are parameterized, and how they can be analyzed to project spread rates and the sensitivity of those rates to changes in model parameters. For illustrative purposes, we apply these models to Cytisus scoparius, a large shrub in the legume family that is considered a noxious invasive species in eastern and western North America, Chile, Australia, and New Zealand. PMID- 15357803 TI - Establishment risks for invasive species. AB - This article presents a quantitative methodology for evaluating the probability of invasive pest species establishing persistent populations. The estimation of pest establishment relies on data and information describing the biology and ecology of the pest and its interactions with potential host species and the regional environment. This information is developed using a model construct borrowed from theoretical population ecology. The methodology for estimating the probability of pest establishment is part of an overall framework that explores the implications of reductions in pest invasions on subsequent establishment. The risk reduction framework integrates the engineering aspects of different technologies for reducing pest entry, the biology and ecology of pest species, the suitability of potentially susceptible hosts, and the quality of available habitats. The methodology for estimating the risk of establishment is presented using an example pest, the Asian longhorned beetle (Anoplophora glabripennis), which has been introduced into the United States via solid wood packing materials (SWPM) used in international commerce. Uncertainties inherent to the estimation of model parameters that determine the risk of establishment are defined, quantified, and propagated through the population model. Advantages and limitations of the proposed methodology are discussed along with recommendations to make the approach more useful in the management of risks posed by the establishment of pest populations. PMID- 15357804 TI - Ecological risk assessment conceptual model formulation for nonindigenous species. AB - This article addresses the application of ecological risk assessment at the regional scale to the prediction of impacts due to invasive or nonindigenous species (NIS). The first section describes risk assessment, the decision-making process, and introduces regional risk assessment. A general conceptual model for the risk assessment of NIS is then presented based upon the regional risk assessment approach. Two diverse examples of the application of this approach are presented. The first example is based upon the dynamics of introduced plasmids into bacteria populations. The second example is the application risk assessment approach to the invasion of a coastal marine site of Cherry Point, Washington, USA by the European green crab. The lessons learned from the two examples demonstrate that assessment of the risks of invasion of NIS will have to incorporate not only the characteristics of the invasive species, but also the other stresses and impacts affecting the region of interest. PMID- 15357805 TI - What can decision analysis do for invasive species management? AB - Decisions about management of invasive species are difficult for all the reasons typically addressed by multiattribute decision analysis: uncertain outcomes, multiple and conflicting objectives, and many interested parties with differing views on both facts and values. This article illustrates how the tools of multiattribute analysis can improve management of invasive species, with an emphasis on making explicit the social values and preferences that must inform invasive species management. Risk assessment protocols developed previously for invasive species management typically suffer from two interacting flaws: (1) separating risk assessment from risk management, thus disrupting essential connections between the social values at stake in invasive species decisions and the scientific knowledge necessary to predict the likely impacts of management actions, and (2) relying on expert judgment about risk framed in qualitative and value-laden terms, inadvertently mixing the expert's judgment about what is likely to happen with personal preferences. Using the values structuring and probability-modeling elements of formal decision analysis can remedy these difficulties and make invasive species management responsive to both good science and public values. The management of feral pigs in Hawaiian ecosystems illustrates the need for such an integrated approach. PMID- 15357806 TI - Habitat destruction, fragmentation, and disturbance promote invasion by habitat generalists in a multispecies metapopulation. AB - Species invasions are extremely common and are vastly outpacing the ability of resource agencies to address each invasion, one species at a time. Management actions that target the whole landscape or ecosystem may provide more cost effective protection against the establishment of invasive species than a species by-species approach. To explore what ecosystem-level actions might effectively reduce invasions, we developed a multispecies, multihabitat metapopulation model. We assume that species that successfully establish themselves outside their native range tend to be habitat generalists and that a tradeoff exists between competitive ability and habitat breadth, such that habitat specialists are competitively superior to habitat generalists. In this model, habitat destruction, fragmentation, and short-term disturbances all favor invasion by habitat generalists, despite the inferior competitive abilities of generalist species. Our model results illustrate that providing relatively undisturbed habitat and preventing further habitat degradation and fragmentation can provide a highly cost-effective defense against invasive species. PMID- 15357807 TI - Bioeconomics of managing the spread of exotic pest species with barrier zones. AB - Exotic pests are serious threats to North American ecosystems; thus, economic analysis of decisions about eradication, stopping, or slowing their spread may be critical to ecosystem management. The proposed bioeconomic model assumes that the rate of population expansion can be reduced (even to negative values in a case of eradication) if certain management actions are taken along the population front. The area of management can be viewed as a dynamic barrier zone that moves together with the population front. The lower is the target rate of spread, the higher would be both benefits and costs of the project. The problem is to find the optimal target rate of spread at which the present value of net benefits from managing population spread reaches its maximum value. If a population spreads along an infinite habitat strip, the target rate of spread is optimal if the slope of the cost function versus the rate of spread is equal to the ratio of the average pest-related damage per unit time and unit area to the discount rate. In a more complex model where the potential area of expansion is limited, two local maxima of net benefits may exist: one for eradication and another for slowing the spread. If both maxima are present, their heights are compared and the strategy that corresponds to a higher value of net benefits is selected. The optimal strategy changes from eradication to slowing the spread and finally to doing nothing as the area occupied by the species increases. The model shows that slowing the spread of pest species generates economic benefits even if a relatively small area remains uninfested. The cost of slowing the spread can be estimated from a model of population expansion via establishment of isolated colonies beyond the moving front. The model is applied to managing the spread of the gypsy moth (Lymantria dispar) populations in the United States. PMID- 15357808 TI - Risk analysis for invasive species: general framework and research needs. AB - A joint workshop was convened by the Society for Risk Analysis Ecological Risk Assessment Specialty Group and the Ecological Society of America Theoretical Ecology Section to provide independent scientific input into the formulation of methods and processes for risk assessment of invasive species. In breakout sessions on (1) the effects of invasive species on human health, (2) effects on plants and animals, (3) risk analysis issues and research needs related to entry and establishment of invasive species, and (4) risk analysis issues and research needs related to the spread and impacts of invasive species, workshop participants discussed an overall approach to risk assessment for invasive species. Workshop participants agreed on the need for empirical research on areas in which data are lacking, including potential invasive species, native species and habitats that may be impacted by invasive species, important biological processes and phenomena such as dispersal, and pathways of entry and spread for invasive species. Participants agreed that theoretical ecology can inform the process of risk assessment for invasive species by providing guidelines and conceptual models, and can contribute to improved decision making by providing a firm biological basis for risk assessments. PMID- 15357809 TI - A discussion of findings and their possible implications from a workshop on bioterrorism threat assessment and risk management. AB - In November 2001, the Monterey Institute of International Studies convened a workshop on bioterrorism threat assessment and risk management. Risk assessment practitioners from various disciplines, but without specialized knowledge of terrorism, were brought together with security and intelligence threat analysts to stimulate an exchange that could be useful to both communities. This article, prepared by a subset of the participants, comments on the workshop's findings and their implications and makes three recommendations, two short term (use of threat assessment methodologies and vulnerability analysis) and one long term (application of quantitative risk assessment and modeling), regarding the practical application of risk assessment methods to bioterrorism issues. PMID- 15357810 TI - CAPPS II: the foundation of aviation security? AB - A new computer system is being developed to classify U.S. air travelers by the degree of terrorist threat they might pose. Reports indicate that the system- called CAPPS II--would use large amounts of information about each passenger, perhaps including such personal details as his or her magazine-subscription behavior. We argue that what is publicly known about CAPPS II raises questions about how substantially the system would improve aviation security. We discuss conditions under which CAPPS II could yield safety benefits, but suggest that it might be more prudent to view the system as one component of future security arrangements rather than the centerpiece of these arrangements. PMID- 15357811 TI - The lamppost, the wizard, and the law: reflections on Professor Barnett's assessment of CAPPS II. PMID- 15357812 TI - CAPPS II: a risky choice concerning an untested risk detection technology. PMID- 15357813 TI - Comments on "CAPPS II: the foundation of aviation security?". PMID- 15357814 TI - Safety versus defense: comments on "CAPPS II: the foundation of aviation security?". PMID- 15357816 TI - The problem of redundancy problem: why more nuclear security forces may produce less nuclear security. PMID- 15357817 TI - Redundancy and nuclear security. PMID- 15357818 TI - Thinking about how many guards will do the job. PMID- 15357819 TI - Redundancy as a design principle and an operating principle. PMID- 15357820 TI - Increasing the number of guards at nuclear power plants. PMID- 15357821 TI - A risk-based approach to setting priorities in protecting bridges against terrorist attacks. AB - This article presents an approach to the problem of terrorism risk assessment and management by adapting the framework of the risk filtering, ranking, and management method. The assessment is conducted at two levels: (1) the system level, and (2) the asset-specific level. The system-level risk assessment attempts to identify and prioritize critical infrastructures from an inventory of system assets. The definition of critical infrastructures offered by Presidential Decision Directive 63 was used to determine the set of attributes to identify critical assets--categorized according to national, regional, and local impact. An example application is demonstrated using information from the Federal Highway Administration National Bridge Inventory for the State of Virginia. Conversely, the asset-specific risk assessment performs an in-depth analysis of the threats and vulnerabilities of a specific critical infrastructure. An illustration is presented to offer some insights in risk scenario identification and prioritization, multiobjective evaluation of management options, and extreme event analysis for critical infrastructure protection. PMID- 15357822 TI - Mathematical methods in combatting terrorism. AB - This document is a preliminary report on the role that mathematical and statistical methods might play in the defense against terrorist attacks. In no way does this replace the efforts of law enforcement agencies or intelligence activities. The hope is that mathematical techniques can make their efforts more efficient. The ideas enumerated here utilize the notion of probabilistic risk analysis, which was developed for the purpose of assessing the safety of nuclear reactors, as well as randomization and game theory. More extensive work in these directions is contemplated for the future. The author is planning workshops to evaluate the ideas presented here and to elicit additional methodologies that may prove useful in this endeavor. PMID- 15357823 TI - Expressing economic risk--review and presentation of a unifying approach. AB - Risk related to economic values is treated by many disciplines, including safety and production engineering, business, and project management. Within each of these and across these disciplines different nomenclature and principles are adopted for describing and communicating risk. The situation is rather confusing. In this article, we review various approaches and concepts that are used to express risk. We present and discuss a unifying approach for dealing with economic risk, with uncertainty being the key risk concept. The approach represents a rethinking on how to implement the Bayesian paradigm in practice to support decision making. PMID- 15357824 TI - The impact of dietary changes among the Inuit of Nunavik (Canada): a socioeconomic assessment of possible public health recommendations dealing with food contamination. AB - Inuit populations meet a large portion of their food needs by eating country food in which pollutants are concentrated. Despite the fact that they contain pollutants, the consumption of country food has many health, social, economic, and cultural benefits. A risk determination process was set up in order to help regional health authorities of Nunavik to deal with this particular issue. Based on Nunavik health authorities' objectives to encourage the region's inhabitants to change their dietary habits, and on both the risks and the benefits of eating country food, several management options were developed. The options aimed at reducing exposure to contaminants by either substituting certain foods with others that have a lower contaminant content or by store-bought foods. This article aims at assessing the potential economic impact of these risk management options before being implemented. Relevant economic data (aggregate income and monetary outlays for the purchase of food and equipment required for food production by households) were collected and identified to serve as a backdrop for the various replacement scenarios. Results show that household budgets, and the regional economy, are not significantly affected by the replacement of contaminated foods with the purchase of store-bought meat, and even less so if the solution involves replacing contaminated foods with other types of game hunted in the region. When financial support is provided by the state, the households can even gain some monetary benefits. Results show that public health authorities' recommended changes to dietary habits among the Inuit of Nunavik would not necessarily involve economic constraints for Inuit households. PMID- 15357825 TI - Quantification of variability and uncertainty for censored data sets and application to air toxic emission factors. AB - Many environmental data sets, such as for air toxic emission factors, contain several values reported only as below detection limit. Such data sets are referred to as "censored." Typical approaches to dealing with the censored data sets include replacing censored values with arbitrary values of zero, one-half of the detection limit, or the detection limit. Here, an approach to quantification of the variability and uncertainty of censored data sets is demonstrated. Empirical bootstrap simulation is used to simulate censored bootstrap samples from the original data. Maximum likelihood estimation (MLE) is used to fit parametric probability distributions to each bootstrap sample, thereby specifying alternative estimates of the unknown population distribution of the censored data sets. Sampling distributions for uncertainty in statistics such as the mean, median, and percentile are calculated. The robustness of the method was tested by application to different degrees of censoring, sample sizes, coefficients of variation, and numbers of detection limits. Lognormal, gamma, and Weibull distributions were evaluated. The reliability of using this method to estimate the mean is evaluated by averaging the best estimated means of 20 cases for small sample size of 20. The confidence intervals for distribution percentiles estimated with bootstrap/MLE method compared favorably to results obtained with the nonparametric Kaplan-Meier method. The bootstrap/MLE method is illustrated via an application to an empirical air toxic emission factor data set. PMID- 15357826 TI - Simple bounds on cumulative intensity functions of renewal and G-renewal processes with increasing failure rate underlying distributions. AB - The article considers point processes most commonly used in reliability and risk analysis. Short-term and long-term behavior for the point processes used as models for repairable systems(1) are introduced. As opposed to the long term, the term short term implies that a process is observed during an interval limited by a time close to the mean (or the median) of the respective underlying distribution. A new simple upper bound is proposed on the cumulative intensity function of the renewal process and G-renewal process with an increasing failure rate underlying distribution. The new bound is compared with some known bounds for the renewal process. Finally, a formal definition of "a boundary point" between the short-term repairable system behavior and long-term behavior is introduced. This point can also be used as a lower time limit beyond which the "long-term" Barlow and Proschan bound for the renewal process with NBUE underlying distribution could be effectively applied. PMID- 15357827 TI - Use of advances in technology for maritime risk assessment. AB - The maritime industry is moving toward a "goal-setting" risk-based regime. This opens the way to safety engineers to explore and exploit flexible and advanced risk modeling and decision-making approaches in the design and operation processes. In this article, following a brief review of the current status of maritime risk assessment, a design/operation selection framework and a design/operation optimization framework are outlined. A general discussion of control engineering techniques and their application to risk modeling and decision making is given. Four novel risk modeling and decision-making approaches are then outlined with illustrative examples to demonstrate their use. Such approaches may be used as alternatives to facilitate risk modeling and decision making in situations where conventional techniques cannot be appropriately applied. Finally, recommendations on further exploitation of advances in general engineering and technology are suggested with respect to risk modeling and decision making. PMID- 15357831 TI - Melatonin, melatonin receptors and melanophores: a moving story. AB - Melatonin (5-methoxy N-acetyltryptamine) is a hormone synthesized and released from the pineal gland at night, which acts on specific high affinity G-protein coupled receptors to regulate various aspects of physiology and behaviour, including circadian and seasonal responses, and some retinal, cardiovascular and immunological functions. In amphibians, such as Xenopus laevis, another role of melatonin is in the control of skin coloration through an action on melanin containing pigment granules (melanosomes) in melanophores. In these cells, very low concentrations of melatonin activate the Mel(1c) receptor subtype triggering movement of granules toward the cell centre thus lightening skin colour. Mel(1c) receptor activation reduces intracellular cAMP via a pertussis toxin-sensitive inhibitory G-protein (Gi), but how this and other intracellular signals regulate pigment movement is not yet fully understood. However, melanophores have proven an excellent model for the study of the molecular mechanisms which coordinate intracellular transport. Melanosome transport is reversible and involves both actin- (myosin V) and microtubule-dependent (kinesin II and dynein) motors. Melanosomes retain both kinesin and dynein during anterograde and retrograde transport, but the myosin V motor seems to be recruited to melanosomes during dispersion, where it assists kinesin II in dominating dynein thus driving net dispersion. Recent work suggests an important role for dynactin in coordinating the activity of the opposing microtubule motors. The melanophore pigment aggregation response has also played a vital role in the ongoing effort to devise specific melatonin receptor antagonists. Much of what has been learnt about the parts of the melatonin molecule required for receptor binding and activation has come from detailed structure-activity data using novel melatonin ligands. Work aiming to devise ligands specific for the distinct melatonin receptor subtypes stands poised to deliver selective agonists and antagonists which will be valuable tools in understanding the role of this enigmatic hormone in health and disease. PMID- 15357832 TI - Functional genomics tools for the analysis of zebrafish pigment. AB - Genetic model organisms are increasingly valuable in the post-genomics era to provide a basis for comparative analysis of the human genome. For higher order processes of vertebrate pigment cell biology and development, the mouse has historically been the model of choice. A complementary organism, the zebrafish (Danio rerio), shares many of the signaling and biological processes of vertebrates, e.g. neural crest development. The zebrafish has a number of characteristics that make it an especially valuable model for the study of pigment cell biology and disease. Large-scale genetic screens have identified a collection of pigmentation mutants that have already made valuable contributions to pigment research. An increasing repertoire of genomic resources such as an expressed sequence tag-based Gene Index (The Institute for Genomic Research) and improving methods of mutagenesis, transgenesis, and gene targeting make zebrafish a particularly attractive model. Morpholino phosphorodiamidate oligonucleotide (MO) 'knockdown' of pigment gene expression provides a non-conventional antisense tool for the analysis of genes involved in pigment cell biology and disease. In addition, an ongoing, reverse-genetic, MO-based screen for the rapid identification of gene function promises to be a valuable complement to other high-throughput microarray and proteomic approaches for understanding pigment cell biology. Novel reagents for zebrafish transgenesis, such as the Sleeping Beauty transposon system, continue to improve the capacity for genetic analysis in this system and ensure that the zebrafish will be a valuable genetic model for understanding a variety of biological processes and human diseases for years to come. PMID- 15357833 TI - Stimulation of cyclic GMP efflux in human melanocytes by hypergravity generated by centrifugal acceleration. AB - Gravity alteration (micro- and hypergravity) is known to influence cell functions. As guanosine 3',5'-cyclic monophosphate (cGMP) plays an important role in human melanocyte functions and different guanylyl cyclase isoforms are responsible for cGMP synthesis in human non-metastatic and metastatic melanoma cells, we investigated the effects of hypergravity on the regulation of cGMP levels in cultured human melanocytes and in melanoma cell lines with different metastatic potentials. Hypergravity was produced by horizontal centrifugal acceleration. Here we report that long-term application of hypergravity (up to 5 g for 24 h) stimulated cGMP efflux in cultured melanocytes and in non-metastatic melanoma cells in the presence of 0.1 mM 3-isobutyl-1-methylxanthine (IBMX), a non-selective phosphodiesterase (PDE) inhibitor. Under these conditions, cAMP synthesis and melanin production were up-regulated in pigmented melanocytes and non-metastatic melanoma cells. Hypergravity also stimulated cGMP transport in the presence of 1 microM trequinsin, an inhibitor of cGMP-binding PDE (PDE5) and of transport by multidrug resistance proteins MRP4/5, whereas 50 microM trequinsin partially inhibited cGMP transport. Transport was further inhibited by probenecid, an inhibitor of endogenous non-selective transporters as well as of MRP4/5 and by cycloheximide as an inhibitor of de novo protein synthesis. In contrast, hypergravity did not affect cGMP efflux in metastatic melanoma cells, which might be related to an up-regulated cGMP efflux at 1 g. The results of the present study indicate that hypergravity may stimulate cGMP efflux in melanocytes and in non-metastatic melanoma cells most probably by an enhanced expression of endogenous transporters and/or MRP4/5. Thus, an altered acceleration vector may induce signaling events in melanocytic cells. PMID- 15357834 TI - The endothelin/sarafotoxin-induced increase of the proliferation of undifferentiated and DMSO-differentiated GEM-81 goldfish erythrophoroma cells is mediated by ETB receptors. AB - Endothelins (ETs) and sarafotoxins (SRTXs) have been reported to exert ET(B) mediated effects on vertebrate pigment cells. GEM-81 cell line, a red pigment cell-derived cutaneous tumor of the teleost Carassius auratus, expresses ET(B) receptors and can be differentiated with 1.5% DMSO treatment, thus constituting an useful model to investigate ET and SRTX effects on cultured fish pigment cells. Our aim was to characterize the pharmacology and biological effects mediated by ET receptors in DMSO-differentiated and undifferentiated cells. ET subtype receptors and their respective Ki values in both cell types were determined by competitive binding assays using (125)I ET-1 and BQ-485 (an ET(A) antagonist) or BQ-788 (an ET(B) antagonist). BQ-788, but not BQ-485, significantly reduced (125)I-ET-1 binding in both cell types, with similar low (Ki > nM) affinities. To determine the proliferation effects of ETs/SRTXs, cells were treated for 72 h with the hormones, and counted in a hemocytometer. The proliferation assays were repeated for SRTX S6c in the presence or absence of BQ 788. The results demonstrated that, with the exception of ET-1 (biphasic effect) and ET-3 (no significant effect) in undifferentiated GEM-81 cells, all the tested hormones induced increases in the proliferation of both types of cells. The hormones were equipotent in DMSO-differentiated cells, which exhibited increased sensitivity to ETs, but not to SRTXs, as compared with undifferentiated cells. The BQ-788 antagonistic effect was also exerted on the proliferation responses to SRTX S6c. These results corroborate the long and important evolutionary history of the ET/SRTX receptor system in vertebrate pigment cells. PMID- 15357835 TI - Absence of TRP-2 in melanogenic melanocytes of human hair. AB - Skin and hair colour mostly depend on the activity of melanogenic melanocytes. Numerous proteins involved in melanocyte function have been identified including pMel-17, Mitf-M, Sox10, tyrosinase, tyrosinase related proteins-1 (TRP-1) and -2 (TRP-2). In the hair, melanogenic activity occurs only during the anagen phase of the hair cycle. In order to evaluate the implications of some known melanogenic proteins in human hair pigmentation, we performed immunohistochemical studies to reveal the expression of pMel-17, Mitf-M, tyrosinase, TRP-1 and TRP-2 in active bulb melanocytes of eumelanic brown and black anagen hairs of different ethnic origins, e.g. brown Caucasian, black Asian and African hairs. The labelling was compared with that observed in Caucasian and African scalp epidermis (interfollicular epidermis) melanocytes. We found that while pMel-17, TRP-1 and TRP-2 were expressed in epidermal melanocytes irrespective of ethnic origin and melanin content of the scalp epidermis, Mitf-M and tyrosinase expression were clearly evidenced only in pigmented epidermis, e.g. African scalps. Regarding human hair, pMel-17, Mitf-M, tyrosinase and TRP-1 were detected in a similar manner in active bulb melanocytes of brown and black hairs. In contrast and unexpectedly, TRP-2 could not be detected in hair bulb melanocytes, whatever the hair colour and ethnic origin. The lack of TRP-2 was further confirmed by western blot analyses. Reverse transcriptase-polymerase chain reaction (RT-PCR) performed on hair bulb mRNA demonstrated that Mitf-M, tyrosinase and TRP-1 amplimer signals were easily detected, whereas the TRP-2 amplimer signal was barely detectable. Furthermore Sox10 was not detected in hair bulb. Altogether our results suggest that the absence of detectable level of TRP-2 is due to transcriptional control in active melanocytes of human eumelanic hair bulbs. According to the absence of TRP-2 in melanin-producing melanocytes of brown and black hair bulbs, one must consider that eumelanogenesis as well as brown and black colour do not require TRP-2 expression in human hair. PMID- 15357836 TI - Rab27b is up-regulated in human Griscelli syndrome type II melanocytes and linked to the actin cytoskeleton via exon F-Myosin Va transcripts. AB - Patients with the autosomal recessive Griscelli-Prunieras syndrome type II are immunologically impaired and have an unusual silvery-grey hypopigmented colour of scalp hair, eyelashes and eyebrows but no noteworthy pigmentary abnormalities of the skin. In most Griscelli patients, the RAB27A gene, which encodes a small GTPase that is associated with the melanosome membrane in melanocytes, is mutated. Here we discuss a genomic RAB27A deletion found in a 21-month-old Moroccan Griscelli patient. Additionally, we provide evidence that the loss of functional Rab27a in melanocytes of this Griscelli patient is partially compensated by the up-regulation of Rab27b, a homologue of Rab27a. By real-time quantitative PCR and western blot analysis, we found that Rab27b mRNA and protein, expressed at low levels in normal human melanocytes, is significantly up regulated in melanocytes derived from this patient. Our immunofluorescence and yeast two-hybrid screening studies reveal that Rab27b can form a tripartite complex on the melanosome membrane with Melanophilin, a Rab27a effector, and protein products of Myosin Va transcripts that contain exon F. Our data suggest that up-regulated Rab27b in melanocytes of the Griscelli patient can partially take over the function of Rab27a, which could explain the fact that this patient had an evenly pigmented skin and was able to tan. PMID- 15357837 TI - Pheomelanin production in the epidermis from newborn agouti mice is induced by the expression of the agouti gene in the dermis. AB - The present study was designed to clarify the role of the agouti gene in the regulation of the proliferation and differentiation of mouse epidermal melanocytes using serum-free primary culture of epidermal melanocytes from 0.5-d old black (a/a; C57BL/10JHir) mice and congenic, agouti (A/A; C57BL/10JHir-A/A) mice. There was no significant difference in the proliferation or differentiation of melanocytes between a/a and A/A mice. However, the content of pheomelanin in culture media from A/A melanocytes was increased by L-tyrosine compared with a/a melanocytes. In addition, the content of the pheomelanin precursor, 5-S cysteinyldopa, in culture media from A/A melanocytes was dramatically increased by L-tyrosine. Moreover, pheomelanin content in the epidermis from 3.5- and 5.5-d old A/A mice was much higher than in a/a mice. Analysis of the A gene using reverse transcription-polymerase chain reaction revealed that cultured keratinocytes and melanocytes do not express the A gene. Moreover, the A gene was expressed in the A/A dermis of 0.5-, 3.5- and 5.5-d-old mice, but not in the a/a dermis nor in the A/A or a/a epidermis. These results suggest that A/A epidermal melanoblasts are influenced by the A gene from the dermis of neonatal mice, and are capable of synthesizing pheomelanin in the culture. Pheomelanin production in the epidermis from 3.5- and 5.5-d-old A/A mice may be induced by the expression of the agouti gene in the dermis. PMID- 15357838 TI - The pigmentation of human iris influences the uptake and storing of zinc. AB - Age-related macular degeneration (AMD) is more prevalent among the elderly Caucasians than in Africans. A significant association between light iris colour, fundus pigmentation and incidence of AMD is reported, suggesting a possible correlation with melanin pigment. Zinc is known to bind to melanin in pigmented tissues and to enhance antioxidant capacity by function as a cofactor or gene expression factor of antioxidant enzymes in the eye. In this in vitro study, we investigated the uptake and storage of zinc in human irides. Irides of blue and brown human eyes were used. The number of melanocytes was measured. Tissues without any treatment served as controls. The irides were incubated with 100 microM zinc chloride in culture medium for 24 h. Specimens of the tissues were stored for the uptake examination. The remained pieces were further incubated for 3 and 7 d to investigate the storage of zinc. The concentration of zinc was measured by inductively coupled plasma mass spectrometry (ICP-MS). Melanocytes count was significantly higher in the brown tissues (P < 0.0001). Zinc concentration of blue coloured irides after 24 h zinc treatment was close to the controls. We did not observe any significant storing. In contrast, the concentration of zinc in brown irides was significantly increased after 24 h (P < or = 0.01) and remained at a high level for 7 d. The uptake of zinc is likely dependent on the amount of pigmentation in human iris. Therefore, we assume that in patients suffering from AMD the degree of pigmentation of the irides and eventually fundi should be under consideration when the patients are treated with zinc supplementation. PMID- 15357839 TI - Direct effects of visible and UVA light on pigment migration in erythrophores of Nile tilapia. AB - Erythrophores derived from Nile tilapia (Oreochromis niloticus) are sensitive to visible light of defined wavelengths in primary culture in the same manner as erythrophores in the skin. Cultured erythrophores aggregate their pigment in response to light with peak wavelengths near 400 or 600 nm, while dispersion is caused by light near 500 nm. In this study, we report that ultraviolet A (UVA) with a peak wavelength near 365 nm also induces pigment aggregation in erythrophores in the skin and in primary culture. The responses of erythrophores in the skin or in culture depend on the light intensity, although the photo sensitivity differs among individual cells. From the results, we conclude that the action of visible light and UVA light on tilapia erythrophores is direct, and that multiple types of visual pigments may coexist in individual erythrophores. PMID- 15357840 TI - Expression of gp100 and CDK2 in melanoma cells is not co-regulated by a shared promoter region. AB - Expression of the pigmentation-associated gene PMel17 is regulated by a 1 kB promoter region shared between the PMel17 and CDK2 genes. The encoded melanosomal glycoprotein gp100 and the cell cycle regulatory protein CDK2 are transcribed in opposite directions. Luciferase reporter constructs were generated for subregions of the promoter containing 0, 1, 2 or 3 putative binding sites for transcription factors with basic helix-loop-helix (bHLH) motifs. The potential contribution of bHLH transcription factor microphthalmia transcription factor (MITF) to promoter activity was investigated by re-introducing microphthalmia into melanoma cells lacking expression. A bi-directional reporter construct was generated to investigate potential co-regulation of gp100 and CDK2 transcription. Promoter activity was assessed in presence and absence of phorbol ester tetradecanoyl phorbol 13-acetate (TPA). FACS analysis and immunohistology served to evaluate co regulation of gp100 and CDK2 expression at the protein level. The full-length promoter, including a consensus binding site for MITF was found to contain sequences that suppressed gp100 expression. Introduction of MITF into non expressing 1123 melanoma cells did not restore gp100 expression levels. A lack of coregulation for gp100 and CDK2 as suggested by immunostaining was supported by findings of dissimilar expression regulation by TPA for either gene. The current study provides insight into transcriptional regulation of the PMel17 and CDK2 genes, important to identify strategies for modulating expression of gp100 and CDK2 proteins by melanoma cells. PMID- 15357841 TI - Melanocyte development: with a message of encouragement to young women scientists. AB - This is a semi-biographical review describing my research on melanocyte development and related personal experiences. Having been educated and trained as a dermatologist, I have been involved in many clinically-oriented studies, however, what has always interested me the most is pigment cell biology. Since I started working at St Marianna University in 1991, I have been undertaking research on melanocyte development and relevant growth factors using mice as models. My research in this field was inspired by my collaborations with various scientists, mostly from the field of biology. Many of these specialists I have met at meetings of the Societies of Pigment Cell Research (PCR). Stem cell factor (SCF, Kitl) and endothelin 3 (EDN3) have been identified as indispensable factors regulating the development of melanocytes. Mice mutant at loci encoding those factors (or their receptors) such as Sl/Sl (receptors W/W) and ls/ls (receptors s/s) have white coat colors and white patches, respectively. Our murine neural crest cell (NCC) primary cultures derived from Sl/Sl embryos showed that EDN3 cannot develop melanocyte precursors without SCF and that EDN3 can elicit proliferation and differentiation in the presence of SCF. These results suggest that without EDN3 and the endothelin type B receptor (EDNRB), melanocytes can not fully increase in number, which could well be the cause of the partial white coat color of ls/ls and s/s mice. Contamination with factors derived from the serum in medium or in feeder cells sometimes causes experimental errors, and therefore we established three immortal cell lines derived from NCC in different developmental stages and designated them as NCCmelb4, NCCmelb4M5 and NCCmelan5, all of which can survive without feeder cells. Using these cell lines and NCC primary cultures, we studied the effect of many factors related to melanocyte development. From the results, it has become evident that Vitamin D3 induces EDNRB expression by NCCmelb4 cells. In addition to the International Pigment Cell Conference (IPCC), I have also taken part in many annual meetings of the Japanese Society for Pigment Cell Research (JSPCR), Pan American Society for Pigment Cell Research (PASPCR) and European Society for Pigment Cell Research (ESPCR). Not only have I learned a great deal, I have enjoyed myself immensely at those meetings. Moreover, I have made many good friends there, some of whom I have collaborated with in my research. To conclude, I would like to give my message 'be ambitious' to young scientists, especially young women. PMID- 15357842 TI - Ultraviolet radiation induces dose-dependent pigment dispersion in crustacean chromatophores. AB - Pigment dispersion in chromatophores as a response to UV radiation was investigated in two species of crustaceans, the crab Chasmagnathus granulata and the shrimp Palaemonetes argentinus. Eyestalkless crabs and shrimps maintained on either a black or a white background were irradiated with different UV bands. In eyestalkless crabs the significant minimal effective dose inducing pigment dispersion was 0.42 J/cm(2) for UVA and 2.15 J/cm(2) for UVB. Maximal response was achieved with 10.0 J/cm(2) UVA and 8.6 J/cm(2) UVB. UVA was more effective than UVB in inducing pigment dispersion. Soon after UV exposure, melanophores once again reached the initial stage of pigment aggregation after 45 min. Aggregated erythrophores of shrimps adapted to a white background showed significant pigment dispersion with 2.5 J/cm(2) UVA and 0.29 J/cm(2) UVC. Dispersed erythrophores of shrimps adapted to a black background did not show any significant response to UVA, UVB or UVC radiation. UVB did not induce any significant pigment dispersion in shrimps adapted to either a white or a black background. As opposed to the tanning response, which only protects against future UV exposure, the pigment dispersion response could be an important agent protecting against the harmful effects of UV radiation exposure. PMID- 15357843 TI - Melanin synthesis may sensitize melanocytes to oxidative DNA damage by ultraviolet A radiation and protect melanocytes from direct DNA damage by ultraviolet B radiation. PMID- 15357846 TI - Objectives of the meeting: think opiates. PMID- 15357847 TI - The opioid system in the gastrointestinal tract. AB - Mu-, delta- and kappa-opioid receptors (ORs) mediate the effects of endogenous opioids and opiate drugs. Here we report (1) the distribution of muOR in the guinea-pig and human gastrointestinal tract in relation to endogenous ligands, to functionally distinct structures in the gut and to deltaOR and kappaOR; and (2) the ligand-induced muOR endocytosis in enteric neurones using in vitro and in vivo models. In the guinea pig, muOR immunoreactivity is confined mainly to the myenteric plexus. MuOR myenteric neurones are most numerous in the small intestine, followed by the stomach and the proximal colon. MuOR immunoreactive fibres are dense in the muscle layer and the deep muscular plexus, where they are in close association with interstitial cells of Cajal. This distribution closely matches the pattern of enkephalin. MuOR enteric neurones comprise functionally distinct populations of neurones of the ascending and descending pathways of the peristaltic reflex. In human gut, muOR immunoreactivity is localized to myenteric and submucosal neurones and to immune cells of the lamina propria. DeltaOR immunoreactivity is located in both plexuses where it is predominantly in varicose fibres in the plexuses, muscle and mucosa, whereas kappaOR immunoreactivity appears to be confined to the myenteric plexus and to bundles of fibres in the muscle. MuOR undergoes endocytosis in a concentration-dependent manner, in vitro and in vivo. Pronounced muOR endocytosis is observed in neurones from animals that underwent abdominal surgery that has been shown to induce delay in gastrointestinal transit. We can conclude that all three ORs are localized to the enteric nervous system with differences among species, and that muOR endocytosis can be utilized as a means to visualize enteric neurones activated by opioids and sites of opioid release. PMID- 15357848 TI - Function of opioids in the enteric nervous system. AB - Alterations in gastrointestinal motility and secretion underlie the constipating action of therapeutically administered opiates. The prototype opiate is morphine, which acts to delay gastric emptying and intestinal transit, to suppress intestinal secretion of water and electrolytes and to suppress transport of bile into the duodenum. The effects of opiates, synthetic opioids and endogenously released opioid peptides on these organ-level gastrointestinal functions reflect actions on electrical and synaptic behaviour of neurones in the enteric nervous system. Adverse effects and positive therapeutic effects of administration of opioid-receptor-blocking drugs on the digestive tract must be understood in the context of the neurophysiology of the enteric nervous system and mechanisms of neural control of gastrointestinal smooth muscle, secretory glands and blood lymphatic vasculature. We review here the integrated systems of physiology and cellular neurobiology that are basic to understanding the actions of opioid agonists and antagonists in the digestive tract. PMID- 15357849 TI - Peripheral opiate action on afferent fibres supplying the rat intestine. AB - The aim of the present study was to examine the sensitivity of mesenteric afferents supplying the rat small intestine to mu-opioid receptor ligands. Mesenteric afferent discharge was recorded electrophysiologically in response to [D-ALA2, N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO; 100 mug kg(-1) i.v.), before and after treatment with the mu-receptor antagonist alvimopan (1 mg kg(-1) i.v.). DAMGO markedly stimulated whole nerve mesenteric afferent discharge (P < 0.05), an effect completely blocked by alvimopan. The response of mesenteric afferents to 2-methyl-5-hydroxytryptamine (30 microg kg(-1) i.v.), bradykinin (0.1-1 microg kg(-1) i.a.) and both low- and high-threshold distension (0-60 mmHg) was unaffected by alvimopan. In chronically vagotomized animals, the low-threshold response to distension was attenuated while the remaining high-threshold response was unaffected by alvimopan. In conclusion, mesenteric afferent fibres are markedly stimulated by mu-opioid receptor agonists, an effect blocked by alvimopan, which may contribute to the gastrointestinal reflex and behavioural responses to opiate treatment or abuse. However, alvimopan did not influence the normal sensitivity of intestinal afferents to chemical and mechanical stimuli that activate different subpopulations of vagal and spinal afferents. Thus, alvimopan may be useful for the treatment of gastrointestinal sequelae following opiate treatment for postoperative or chronic pain. PMID- 15357850 TI - The role of endogenous opioids in the control of gastrointestinal motility: predictions from in vitro modelling. AB - Gastrointestinal motility can be assessed in vitro by investigating the effects of drugs or gene knockouts on intestinal propulsion, and on neurone-mediated responses evoked by electrical field stimulation (EFS). The latter predominantly measure enteric motor activity and can detect prokinetic activity of exogenous agents. Some evidence suggests that naloxone has prokinetic activity when evaluated for an ability to modulate responses to EFS, but the effects are inconsistent across different species or intestinal regions. Models of intestinal peristalsis measure an integrated sensory-motor nerve function and possess more intact neuro-neuronal connections. In such preparations, the effects of naloxone also suggest a prokinetic property but again, this is inconsistent. By contrast, consistent prokinetic activity of naloxone is apparent in models where peristalsis is compromised by drug-induced suppression of motor nerve activity or by modulation of endogenous processes using receptor antagonists or inappropriate intraluminal distension. These data suggest that endogenous opioids play little or no role in normal intestinal physiology, but suppress intestinal motility when motor function is compromised. Consequently, drugs that antagonize opioid receptors may exert prokinetic activity in conditions where intestinal motility is reduced, such as constipation. Further work is required to elucidate the opiate receptor(s) involved. PMID- 15357851 TI - Preclinical studies of opioids and opioid antagonists on gastrointestinal function. AB - Opioid receptors in the gastrointestinal (GI) tract mediate the effects of endogenous opioid peptides and exogenously administered opioid analgesics, on a variety of physiological functions associated with motility, secretion and visceral pain. The studies reviewed or reported here describe a range of in vivo activities of opioid receptor antagonists upon GI function in rodents, focusing on mu receptors. Naloxone, and the peripherally acting mu-opioid receptor antagonists alvimopan and methylnaltrexone, reverse morphine-induced inhibition of GI transit in mice and rats, and morphine- or loperamide-induced inhibition of castor oil-induced diarrhoea in mice. At doses producing maximal reversal of morphine-induced effects upon GI transit, only the central nervous system (CNS) penetrant antagonist naloxone was able to reverse morphine-induced analgesia. Both central and peripheral opioid antagonists may affect GI function and/or visceromotor sensitivity in the absence of exogenous opioid analgesics, suggesting a constitutive role for endogenous opioid peptides in the control of GI physiology. Furthermore, in contrast to naloxone, alvimopan does not produce hypersensitivity to the visceromotor response induced by nociceptive levels of colorectal distension in a rodent model of post-inflammatory colonic hypersensitivity, suggesting that in the periphery endogenous mu-opioid receptor mediated mechanisms do not regulate colonic sensitivity. The data support the hypothesis that peripherally acting opioid antagonists may be able to selectively block opioid receptors in the GI tract, thereby preserving normal GI physiology, while not blocking the effects of endogenous opioid peptides or exogenous opioid analgesics in the CNS. These findings suggest that the primary sites of action of mu-opioid agonists with respect to inhibition of GI function are in the periphery, whereas analgesic activity resides primarily in the CNS. PMID- 15357852 TI - Mechanisms of postoperative ileus. AB - Postoperative ileus is an iatrogenic condition that follows abdominal surgery. Three main mechanisms are involved in its causation, namely neurogenic, inflammatory and pharmacological mechanisms. In the acute postoperative phase, mainly spinal and supraspinal adrenergic and non-adrenergic pathways are activated. Recent studies, however, show that the prolonged phase of postoperative ileus is caused by an enteric molecular inflammatory response and the subsequent recruitment of leucocytes into the muscularis of the intestinal segments manipulated during surgery. This inflammation impairs local neuromuscular function and activates neurogenic inhibitory pathways, inhibiting motility of the entire gastrointestinal tract. The mechanisms underlying the recruitment of the inflammatory cells, and their interaction with the intestinal afferent innervation, are discussed. Finally, opioids administered for postoperative pain control also contribute to a large extent to the reduction in propulsive gastrointestinal motility observed after abdominal surgery. PMID- 15357853 TI - Clinical perspective on postoperative ileus and the effect of opiates. AB - Postoperative ileus (POI) is a mandatory period of cessation of intestinal function, after abdominal surgery. Over the last decade research has continued into this field, and while we continue to learn about pathogenesis, few controlled data are available for management. The clinical manifestations of POI are reviewed. Management techniques are also reviewed, including the use of postoperative care plans with early ambulation and diet, the effects of epidural anaesthesia, benefits of laparoscopic approaches for intestinal resection, and potential avenues for prevention using novel medications. PMID- 15357855 TI - Proceedings of the Consensus Conference on Hepatitis C. April 21-22, 2004. Edinburgh, United Kingdom. PMID- 15357854 TI - Opiates in the control of gastrointestinal tract function: current knowledge and new avenues for research. PMID- 15357857 TI - Hepatitis C: What is the nature of the problem? AB - This review concentrates on the natural history of the infection. The virology and epidemiology of the disease are covered elsewhere. There are a number of inherent difficulties in studying the outcome of HCV infection: the onset is often not recognized, it can be asymptomatic, the progression is slow and patients may have had treatment. However some insights in recent years have been made. PMID- 15357858 TI - Hepatitis C: who is at risk and how do we identify them? AB - The transmission of, and screening for, HCV infection varies considerably throughout the world; differences between resource-poor and resource-rich countries are particularly pronounced. The perspective of this review, principally, is that of resource-rich countries. The UK, particularly Scotland, experience is drawn on. PMID- 15357859 TI - Hepatitis C: How should we manage the patient? AB - The patient with HCV infection may present with a variety of problems and range from the asymptomatic patient with mild liver damage to a patient presenting with complications of cirrhosis or hepatocellular carcinoma. The diagnosis of hepatitis C may be a complete surprise to the patient or be an expected diagnosis in someone with known risk factors. Similarly the physician may be faced with a patient who knows very little about hepatitis C or someone who has read extensively on the subject. The initial consultation is useful for gaining information on the patient's background, physical examination may give useful clinical clues on the stage of the liver disease. The consultation gives the physician a chance to educate the patient on the current thinking on hepatitis C and to organize confirmatory and other investigations that will help decide on the next line of management, i.e. whether the patient is a candidate for combination therapy of interferon and ribavirin. PMID- 15357860 TI - Hepatitis C: What is the best treatment? AB - The treatment of hepatitis C is expensive, difficult and arduous from the patient's perspective. It is similarly difficult for the clinician to decide who and when to treat. If hepatitis C is viewed from the liver's perspective we need only treat those patients who will develop the complications of chronic liver disease within their lifetimes. If we take a more holistic approach, then we have to consider the implications of being a carrier of a potentially transmissible blood borne virus on the patient themselves, their relationships, their families and their sense of wellbeing. There is now evidence of the large impact HCV has on quality of life and we have to consider extra hepatic manifestations of hepatitis C infection, possibly including the syndrome of 'brain fog' recently described. An additional factor that has to be considered in the decision to treat is whether patients perceive hepatitis C as a significant problem for themselves. For some patients, who have chaotic live styles, it is extremely difficult to get them to access healthcare. To then undergo the rigors and tribulations of hepatitis C therapy that is posing no current problem is unlikely to succeed. However, failure to engage these patients with therapy will lead to a significant proportion of them presenting with serious complications of chronic liver disease, with its attendant mortality, morbidity and cost. Underlying all these considerations is the tension between the costs of therapy and the benefits of therapy. Good arguments can be made in terms of cost-effectiveness for treating patients with a high likelihood of progressing to chronic liver disease and its complications. These arguments become much less persuasive when all patients are concerned. PMID- 15357861 TI - Hepatitis C: What lies ahead and can we afford it? AB - Hepatitis C is a chronic disease with a slow and variable progression over 20-50 years and it is an important public health problem for the 21st century. This paper describes the information required to estimate what lies ahead in terms of morbidity, mortality and the implications for the health service in Scotland and summarises work undertaken in other countries. There will be an increasing number of people with severe liver disease in the next 10-20 years and we need to invest now in primary prevention and effective treatment strategies to reduce the burden of disease in the future. PMID- 15357863 TI - WHO systematic review of maternal morbidity and mortality: the prevalence of severe acute maternal morbidity (near miss). AB - AIM: To determine the prevalence of severe acute maternal morbidity (SAMM) worldwide (near miss). METHOD: Systematic review of all available data. The methodology followed a pre-defined protocol, an extensive search strategy of 10 electronic databases as well as other sources. Articles were evaluated according to specified inclusion criteria. Data were extracted using data extraction instrument which collects additional information on the quality of reporting including definitions and identification of cases. Data were entered into a specially constructed database and tabulated using SAS statistical management and analysis software. RESULTS: A total of 30 studies are included in the systematic review. Designs are mainly cross-sectional and 24 were conducted in hospital settings, mostly teaching hospitals. Fourteen studies report on a defined SAMM condition while the remainder use a response to an event such as admission to intensive care unit as a proxy for SAMM. Criteria for identification of cases vary widely across studies. Prevalences vary between 0.80% - 8.23% in studies that use disease-specific criteria while the range is 0.38% - 1.09% in the group that use organ-system based criteria and included unselected group of women. Rates are within the range of 0.01% and 2.99% in studies using management-based criteria. It is not possible to pool data together to provide summary estimates or comparisons between different settings due to variations in case identification criteria. Nevertheless, there seems to be an inverse trend in prevalence with development status of a country. CONCLUSION: There is a clear need to set uniform criteria to classify patients as SAMM. This standardisation could be made for similar settings separately. An organ-system dysfunction/failure approach is the most epidemiologically sound as it is least open to bias, and thus could permit developing summary estimates. PMID- 15357864 TI - Reproductive health research challenges. PMID- 15357865 TI - Effectiveness and acceptability of progestogens in combined oral contraceptives - a systematic review. AB - BACKGROUND: The progestogen component of oral contraceptives (OCs) has undergone changes since it was recognized that their chemical structure can influence the spectrum of minor adverse and beneficial effects. METHODS: The objective of this review was to evaluate currently available low-dose OCs containing ethinylestradiol and different progestogens in terms of contraceptive effectiveness, cycle control, side effects and continuation rates. The Cochrane Controlled Trials Register, MEDLINE and EMBASE databases were searched. Randomized trials reporting clinical outcomes were considered for inclusion and were assessed for methodological quality and validity. RESULTS: Twenty-two trials were included in the review. Eighteen were sponsored by pharmaceutical companies and in only 5 there was an attempt for blinding. Most comparisons between different interventions included one to three trials, involving usually less than 500 women. Discontinuation was less with second-generation progestogens compared to first-generation (RR 0.79; 95% CI 0.69-0.91). Cycle control appeared to be better with second-compared to first-generation progestogens for both, mono-and triphasic preparations (RR 0.69; 95% CI 0.52-0.91) and (RR 0.61; 95% CI 0.43 0.85), respectively. Intermenstrual bleeding was less with third- compared to second-generation pills (RR 0.71; 95% CI 0.55-0.91).Contraceptive effectiveness of gestodene (GSD) was comparable to that of levonorgestrel (LNG), and had similar pattern of spotting, breakthrough bleeding and absence of withdrawal bleeding). Drospirenone (DRSP) was similar compared to desogestrel (DSG) regarding contraceptive effectiveness, cycle control and side effects. CONCLUSION: The third- and second-generation progestogens are preferred over first generation in all indices of acceptability. Current evidence suggests that GSD is comparable to LNG in terms of contraceptive effectiveness and for most cycle control indices. GSD is also comparable to DSG. DRSP is comparable to DSG. Future research should focus on independently conducted well designed randomized trials comparing particularly the third- with second-generation progestogens. PMID- 15357866 TI - Retroviruses 2004: review of the 2004 Cold Spring Harbor Retroviruses Conference. AB - For the past several decades, retrovirologists from around the world have gathered in late May at the Cold Spring Harbor Laboratories in New York to present their studies in formal talks and posters, and to discuss their ongoing research informally at the bar or on the beach. As organizers of the 2004 Cold Spring Harbor Retroviruses Conference, we have been asked by the editors of Retrovirology to prepare a review of the meeting for publication on-line. Our goal in this review is not to provide a detailed description of data presented at the meeting but rather to highlight some of the significant developments reported this year. The review is structured in a manner that parallels the organization of the meeting; beginning with the entry phase of the replication cycle, proceeding with post-entry events, assembly and release, integration, reverse transcription, pathogenesis/host factors, RNA-related events (transcription, processing, export, and packaging) and finishing with antivirals. While the most striking developments this year involved post-entry events and assembly/release, significant progress was made towards elucidating a number of aspects of the retroviral replication cycle. PMID- 15357867 TI - Co-administration of a DNA vaccine encoding the prostate specific membrane antigen and CpG oligodeoxynucleotides suppresses tumor growth. AB - BACKGROUND: Prostate-specific membrane antigen (PSMA) is a well characterized prostate-specific tumor associated antigen. Its expression is elevated in prostate carcinoma, particularly in metastatic and recurrent lesions. These observations suggest that PSMA can be used as immune target to induce tumor cell specific recognition by the host and, consequently tumor rejection. We utilized a DNA-based vaccine to specifically enhance PSMA expression. An immune modulator, such as CpG oligodeoxynucleotides which promote Th1-type immune responses was combined to increase the efficacy of tumor recognition and elimination. METHODS: A eukaryotic expression plasmid pCDNA3.1-PSMA encoding full-length PSMA was constructed. C57BL/6 mice were immunized with endotoxin-free pCDNA3.1-PSMA alone or in combination with CpG oligodeoxynucleotides by intramuscular injection. After 4 immunizations, PSMA specific antibodies and cytotoxic T lymphocyte reactivity were measured. Immunized C57BL/6 mice were also challenged subcutaneously with B16 cells transfected with PSMA to evaluate suppression of tumor growth. RESULTS: Vaccine-specific cytotoxic T lymphocytes reactive with B16 cells expressing PSMA could be induced with this treatment schedule. Immune protection was observed in vaccinated mice as indicated by increased tumor growth in the control group (100%) compared with the groups vaccinated with DNA alone (66.7%) or DNA plus CpG oligodeoxynucleotides (50%) respectively. Average tumor volume was smaller in vaccinated groups and tumor-free survival time was prolonged by the vaccination. CONCLUSION: The current findings suggest that specific anti-tumor immune response can be induced by DNA vaccines expressing PSMA. In addition, the suppression of in vivo growth of tumor cells expressing PSMA was augmented by CpG oligodeoxynucleotides. This strategy may provide a new venue for the treatment of carcinoma of prostate after failure of standard therapy. PMID- 15357868 TI - Optimisation of the two-dimensional gel electrophoresis protocol using the Taguchi approach. AB - BACKGROUND: Quantitative proteomic analyses have traditionally used two dimensional gel electrophoresis (2DE) for separation and characterisation of complex protein mixtures. Among the difficulties associated with this approach is the solubilisation of protein mixtures for isoelectric focusing (IEF). To find the optimal formulation of the multi-component IEF rehydration buffer (RB) we applied the Taguchi method, a widely used approach for the robust optimisation of complex industrial processes, to determine optimal concentrations for the detergents, carrier ampholytes and reducing agents in RB for 2DE using commercially supplied immobilised pH gradient (IPG) gel strips. RESULTS: Our optimisation resulted in increased protein solubility, improved resolution and reproducibility of 2D gels, using a wide variety of samples. With the updated protocol we routinely detected approximately 4-fold more polypeptides on samples containing complex protein mixtures resolved on small format 2D gels. In addition the pI and size ranges over which proteins could be resolved was substantially improved. Moreover, with improved sample loading and resolution, analysis of individual spots by immunoblotting and mass spectrometry revealed previously uncharacterised posttranscriptional modifications in a variety of chromatin proteins. CONCLUSIONS: While the optimised RB (oRB) is specific to the gels and analysis approach we use, our use of the Taguchi method should be generally applicable to a broad range of electrophoresis and analysis systems. PMID- 15357869 TI - A novel method of cultivating cardiac myocytes in agarose microchamber chips for studying cell synchronization. AB - We have developed a new method that enables agar microstructures to be used to cultivate cardiac myocyte cells in a manner that allows their connection patterns to be controlled. Non-contact three-dimensional photo-thermal etching with a 1064 nm infrared focused laser beam was used to form the shapes of agar microstructures. This wavelength was selected as it is not absorbed by water or agar. Identical rat cardiac myocytes were cultured in adjacent microstructures connected by microchannels and the interactions of asynchronous beating cardiac myocyte cells observed. Two isolated and independently beating cardiac myocytes were shown to form contacts through the narrow microchannels and by 90 minutes had synchronized their oscillations. This occurred by one of the two cells stopping their oscillation and following the pattern of the other cell. In contrast, when two sets of synchronized beating cells came into contact, those two sets synchronized without any observable interruptions to their rhythms. The results indicate that the synchronization process of cardiac myocytes may be dependent on the community size and network pattern of these cells. PMID- 15357870 TI - Immunological evaluation of the new stable ultrasound contrast agent LK565: a phase one clinical trial. AB - BACKGROUND: Ultrasound contrast agents (UCAs) allow the enhancement of vascular definition, thereby providing more diagnostic information. LK565 is a new second generation UCA based on synthetic polymers of aspartic acid which is eliminated from the blood stream via phagocytosis. LK565 forms very stable air-filled microspheres and is capable of repeated passage through the pulmonary capillary bed after peripheral intravenous injection. This characteristic allows examination of the cardiac function or extracardiac vessel abnormalities up to 15 minutes. METHODS: A phase one clinical study was conducted on 15 healthy volunteers to identify the development of an undesirable immune response. Phagocytosis capacity, TNF-alpha secretion, and MHC class II upregulation of monocytes was monitored, as well as microsphere specific antibody development (IgM, IgG). Furthermore, the kinetics of the activation surface markers CD69, CD25, CD71, and CD11b on leukocytes were analyzed. RESULTS: Due to LK565 metabolism the administration of the UCA led to saturation of phagocytes which was reversible after 24 hrs. Compared to positive controls neither significant TNF-alpha elevation, neither MHC class II and activation surface markers upregulation, nor specific antibody development was detectable. CONCLUSION: The administration of LK565 provides a comfortable duration of signal enhancement, esp. in echocardiography, without causing a major activation cascade or triggering an adaptive immune response. To minimize the risk of undesirable adverse events such as anaphylactoid reactions, immunological studies should be included in clinical trials for new UCAs. The use of LK565 as another new ultrasound contrast agent should be encouraged as a safe means to provide additional diagnostic information. PMID- 15357871 TI - Mitoxantrone pleurodesis to palliate malignant pleural effusion secondary to ovarian cancer. AB - BACKGROUND: Advanced ovarian cancer is the leading non-breast gynaecologic cause of malignant pleural effusion. Aim of this study was to assess the efficacy of mitoxantrone sclerotherapy as a palliative treatment of malignant pleural effusions due to ovarian cancer. METHODS: Sixty women with known ovarian cancer and malignant recurrent symptomatic pleural effusion were treated with chest tube drainage followed by intrapleural mitoxantrone sclerotherapy. Survival, complications and response to pleurodesis were recorded. The data are expressed as the mean +/- SEM and the median. RESULTS: The mean age of the entire group was 64 +/- 11,24 years. The mean interval between diagnosis of ovarian cancer and presentation of the effusion was 10 +/- 2,1 months. Eighteen patients (30%) had pleural effusion as the first evidence of recurrence. The mean volume of effusion drained was 1050 +/- 105 ml and chest tube was removed within 4 days in 75% of patients. There were no deaths related to the procedure. Side effects of chemical pleurodesis included fever (37-38,5 degrees C) chest pain, nausea and vomiting. At 30 days among 60 treated effusions, there was an 88% overall response rate, including 41 complete responses and 12 partial responses. At 60 days the overall response was 80% (38 complete responses and 10 partial responses). The mean survival of the entire population was 7,5 +/- 1,2 months. CONCLUSIONS: Mitoxantrone is effective in the treatment of malignant pleural effusion secondary to ovarian cancer without causing significant local or systemic toxicity. PMID- 15357872 TI - A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods. AB - BACKGROUND: Cell migration is a complex phenomenon that requires the coordination of numerous cellular processes. Investigation of cell migration and its underlying biology is of interest to basic scientists and those in search of therapeutics. Current migration assays for screening small molecules, siRNAs, or other perturbations are difficult to perform in parallel at the scale required to screen large libraries. RESULTS: We have adapted the commonly used scratch wound healing assay of tissue-culture cell monolayers to a 384 well plate format. By mechanically scratching the cell substrate with a pin array, we are able to create characteristically sized wounds in all wells of a 384 well plate. Imaging of the healing wounds with an automated fluorescence microscope allows us to distinguish perturbations that affect cell migration, morphology, and division. Readout requires ~1 hr per plate but is high in information content i.e. high content. We compare readouts using different imaging technologies, automated microscopy, scanners and a fluorescence macroscope, and evaluate the trade-off between information content and data acquisition rate. CONCLUSIONS: The adaptation of a wound healing assay to a 384 well format facilitates the study of aspects of cell migration, tissue reorganization, cell division, and other processes that underlie wound healing. This assay allows greater than 10,000 perturbations to be screened per day with a quantitative, high-content readout, and can also be used to characterize small numbers of perturbations in detail. PMID- 15357873 TI - Collecting duct carcinoma of the kidney: an immunohistochemical study of 11 cases. AB - BACKGROUND: Collecting duct carcinoma (CDC) is a rare but very aggressive variant of kidney carcinoma that arises from the epithelium of Bellini's ducts, in the distal portion of the nephron. In order to gain an insight into the biology of this tumor we evaluated the expression of five genes involved in the development of renal cancer (FEZ1/LZTS1, FHIT, TP53, P27kip1, and BCL2). METHODS: We studied eleven patients who underwent radical nephrectomy for primary CDC. All patients had an adequate clinical follow-up and none of them received any systemic therapy before surgery. The expression of the five markers for tumor initiation and/or progression were assessed by immunohistochemistry and correlated to the clinicopathological parameters, and survival by univariate analysis. RESULTS: Results showed that Fez1 protein expression was undetectable or substantially reduced in 7 of the 11 (64%) cases. Fhit protein was absent in three cases (27%). The overexpression of p53 protein was predominantly nuclear and detected in 4 of 11 cases (36%). Immunostaining for p27 was absent in 5 of 11 cases (45.5%). Five of the six remaining cases (90%) showed exclusively cytoplasmic protein expression, where, in the last case, p27 protein was detected in both nucleus and cytoplasm. Bcl2 expression with 100% of the tumor cells positive was observed in 4 of 11 (36%) cases. Statistical analysis showed a statistical trend (P = 0.06) between loss and reduction of Fez1 and presence of lymph node metastases. CONCLUSIONS: These findings suggest that Fez1 may represent not only a molecular diagnostic marker but also a prognostic marker in CDC. PMID- 15357874 TI - A human in vitro model system for investigating genome-wide host responses to SARS coronavirus infection. AB - BACKGROUND: The molecular basis of severe acute respiratory syndrome (SARS) coronavirus (CoV) induced pathology is still largely unclear. Many SARS patients suffer respiratory distress brought on by interstitial infiltration and frequently show peripheral blood lymphopenia and occasional leucopenia. One possible cause of this could be interstitial inflammation, following a localized host response. In this study, we therefore examine the immune response of SARS CoV in human peripheral blood mononuclear cells (PBMCs) over the first 24 hours. METHODS: PBMCs from normal healthy donors were inoculated in vitro with SARS-CoV and the viral replication kinetics was studied by real-time quantitative assays. SARS-CoV specific gene expression changes were examined by high-density oligonucleotide array analysis. RESULTS: We observed that SARS-CoV was capable of infecting and replicating in PBMCs and the kinetics of viral replication was variable among the donors. SARS-CoV antibody binding assays indicated that SARS specific antibodies inhibited SARS-CoV viral replication. Array data showed monocyte-macrophage cell activation, coagulation pathway upregulation and cytokine production together with lung trafficking chemokines such as IL8 and IL17, possibly activated through the TLR9 signaling pathway; that mimicked clinical features of the disease. CONCLUSIONS: The identification of human blood mononuclear cells as a direct target of SARS-CoV in the model system described here provides a new insight into disease pathology and a tool for investigating the host response and mechanisms of pathogenesis. PMID- 15357875 TI - A method for detecting and correcting feature misidentification on expression microarrays. AB - BACKGROUND: Much of the microarray data published at Stanford is based on mouse and human arrays produced under controlled and monitored conditions at the Brown and Botstein laboratories and at the Stanford Functional Genomics Facility (SFGF). Nevertheless, as large datasets based on the Stanford Human array began to accumulate, a small but significant number of discrepancies were detected that required a serious attempt to track down the original source of error. Due to a controlled process environment, sufficient data was available to accurately track the entire process leading to up to the final expression data. In this paper, we describe our statistical methods to detect the inconsistencies in microarray data that arise from process errors, and discuss our technique to locate and fix these errors. RESULTS: To date, the Brown and Botstein laboratories and the Stanford Functional Genomics Facility have together produced 40,000 large-scale (10-50,000 feature) cDNA microarrays. By applying the heuristic described here, we have been able to check most of these arrays for misidentified features, and have been able to confidently apply fixes to the data where needed. Out of the 265 million features checked in our database, problems were detected and corrected on 1.3 million of them. CONCLUSION: Process errors in any genome scale high throughput production regime can lead to subsequent errors in data analysis. We show the value of tracking multi-step high throughput operations by using this knowledge to detect and correct misidentified data on gene expression microarrays. PMID- 15357876 TI - Gene family evolution: an in-depth theoretical and simulation analysis of non linear birth-death-innovation models. AB - BACKGROUND: The size distribution of gene families in a broad range of genomes is well approximated by a generalized Pareto function. Evolution of ensembles of gene families can be described with Birth, Death, and Innovation Models (BDIMs). Analysis of the properties of different versions of BDIMs has the potential of revealing important features of genome evolution. RESULTS: In this work, we extend our previous analysis of stochastic BDIMs. In addition to the previously examined rational BDIMs, we introduce potentially more realistic logistic BDIMs, in which birth/death rates are limited for the largest families, and show that their properties are similar to those of models that include no such limitation. We show that the mean time required for the formation of the largest gene families detected in eukaryotic genomes is limited by the mean number of duplications per gene and does not increase indefinitely with the model degree. Instead, this time reaches a minimum value, which corresponds to a non-linear rational BDIM with the degree of approximately 2.7. Even for this BDIM, the mean time of the largest family formation is orders of magnitude greater than any realistic estimates based on the timescale of life's evolution. We employed the embedding chains technique to estimate the expected number of elementary evolutionary events (gene duplications and deletions) preceding the formation of gene families of the observed size and found that the mean number of events exceeds the family size by orders of magnitude, suggesting a highly dynamic process of genome evolution. The variance of the time required for the formation of the largest families was found to be extremely large, with the coefficient of variation >> 1. This indicates that some gene families might grow much faster than the mean rate such that the minimal time required for family formation is more relevant for a realistic representation of genome evolution than the mean time. We determined this minimal time using Monte Carlo simulations of family growth from an ensemble of simultaneously evolving singletons. In these simulations, the time elapsed before the formation of the largest family was much shorter than the estimated mean time and was compatible with the timescale of evolution of eukaryotes. CONCLUSIONS: The analysis of stochastic BDIMs presented here shows that non-linear versions of such models can well approximate not only the size distribution of gene families but also the dynamics of their formation during genome evolution. The fact that only higher degree BDIMs are compatible with the observed characteristics of genome evolution suggests that the growth of gene families is self-accelerating, which might reflect differential selective pressure acting on different genes. PMID- 15357877 TI - GDR (Genome Database for Rosaceae): integrated web resources for Rosaceae genomics and genetics research. AB - BACKGROUND: Peach is being developed as a model organism for Rosaceae, an economically important family that includes fruits and ornamental plants such as apple, pear, strawberry, cherry, almond and rose. The genomics and genetics data of peach can play a significant role in the gene discovery and the genetic understanding of related species. The effective utilization of these peach resources, however, requires the development of an integrated and centralized database with associated analysis tools. DESCRIPTION: The Genome Database for Rosaceae (GDR) is a curated and integrated web-based relational database. GDR contains comprehensive data of the genetically anchored peach physical map, an annotated peach EST database, Rosaceae maps and markers and all publicly available Rosaceae sequences. Annotations of ESTs include contig assembly, putative function, simple sequence repeats, and anchored position to the peach physical map where applicable. Our integrated map viewer provides graphical interface to the genetic, transcriptome and physical mapping information. ESTs, BACs and markers can be queried by various categories and the search result sites are linked to the integrated map viewer or to the WebFPC physical map sites. In addition to browsing and querying the database, users can compare their sequences with the annotated GDR sequences via a dedicated sequence similarity server running either the BLAST or FASTA algorithm. To demonstrate the utility of the integrated and fully annotated database and analysis tools, we describe a case study where we anchored Rosaceae sequences to the peach physical and genetic map by sequence similarity. CONCLUSIONS: The GDR has been initiated to meet the major deficiency in Rosaceae genomics and genetics research, namely a centralized web database and bioinformatics tools for data storage, analysis and exchange. GDR can be accessed at http://www.genome.clemson.edu/gdr/. PMID- 15357878 TI - Cross-species comparison significantly improves genome-wide prediction of cis regulatory modules in Drosophila. AB - BACKGROUND: The discovery of cis-regulatory modules in metazoan genomes is crucial for understanding the connection between genes and organism diversity. It is important to quantify how comparative genomics can improve computational detection of such modules. RESULTS: We run the Stubb software on the entire D. melanogaster genome, to obtain predictions of modules involved in segmentation of the embryo. Stubb uses a probabilistic model to score sequences for clustering of transcription factor binding sites, and can exploit multiple species data within the same probabilistic framework. The predictions are evaluated using publicly available gene expression data for thousands of genes, after careful manual annotation. We demonstrate that the use of a second genome (D. pseudoobscura) for cross-species comparison significantly improves the prediction accuracy of Stubb, and is a more sensitive approach than intersecting the results of separate runs over the two genomes. The entire list of predictions is made available online. CONCLUSION: Evolutionary conservation of modules serves as a filter to improve their detection in silico. The future availability of additional fruitfly genomes therefore carries the prospect of highly specific genome-wide predictions using Stubb. PMID- 15357879 TI - DIALIGN P: fast pair-wise and multiple sequence alignment using parallel processors. AB - BACKGROUND: Parallel computing is frequently used to speed up computationally expensive tasks in Bioinformatics. RESULTS: Herein, a parallel version of the multi-alignment program DIALIGN is introduced. We propose two ways of dividing the program into independent sub-routines that can be run on different processors: (a) pair-wise sequence alignments that are used as a first step to multiple alignment account for most of the CPU time in DIALIGN. Since alignments of different sequence pairs are completely independent of each other, they can be distributed to multiple processors without any effect on the resulting output alignments. (b) For alignments of large genomic sequences, we use a heuristics by splitting up sequences into sub-sequences based on a previously introduced anchored alignment procedure. For our test sequences, this combined approach reduces the program running time of DIALIGN by up to 97%. CONCLUSIONS: By distributing sub-routines to multiple processors, the running time of DIALIGN can be crucially improved. With these improvements, it is possible to apply the program in large-scale genomics and proteomics projects that were previously beyond its scope. PMID- 15357880 TI - Functionally specified protein signatures distinctive for each of the different blue copper proteins. AB - BACKGROUND: Proteins having similar functions from different sources can be identified by the occurrence in their sequences, a conserved cluster of amino acids referred to as pattern, motif, signature or fingerprint. The wide usage of protein sequence analysis in par with the growth of databases signifies the importance of using patterns or signatures to retrieve out related sequences. Blue copper proteins are found in the electron transport chain of prokaryotes and eukaryotes. The signatures already existing in the databases like the type 1 copper blue, multiple copper oxidase, cyt b/b6, photosystem 1 psaA&B, psaG&K, and reiske iron sulphur protein are not specified signatures for blue copper proteins as the name itself suggests. Most profile and motif databases strive to classify protein sequences into a broad spectrum of protein families. This work describes the signatures designed based on the copper metal binding motifs in blue copper proteins. The common feature in all blue copper proteins is a trigonal planar arrangement of two nitrogen ligands [each from histidine] and one sulphur containing thiolate ligand [from cysteine], with strong interactions between the copper center and these ligands. RESULTS: Sequences that share such conserved motifs are crucial to the structure or function of the protein and this could provide a signature of family membership. The blue copper proteins chosen for the study were plantacyanin, plastocyanin, cucumber basic protein, stellacyanin, dicyanin, umecyanin, uclacyanin, cusacyanin, rusticyanin, sulfocyanin, halocyanin, azurin, pseudoazurin, amicyanin and nitrite reductase which were identified in both eukaryotes and prokaryotes. ClustalW analysis of the protein sequences of each of the blue copper proteins was the basis for designing protein signatures or peptides. The protein signatures and peptides identified in this study were designed involving the active site region involving the amino acids bound to the copper atom. It was highly specific for each kind of blue copper protein and the false picks were minimized. The set of signatures designed specifically for the BCP's was entirely different from the existing broad spectrum signatures as mentioned in the background section. CONCLUSIONS: These signatures can be very useful for the annotation of uncharacterized proteins and highly specific to retrieve blue copper protein sequences of interest from the non redundant databases containing a large deposition of protein sequences. PMID- 15357881 TI - Adaptation of an amphibian mucociliary clearance model to evaluate early effects of tobacco smoke exposure. AB - RATIONALE: Inhaled side-stream tobacco smoke brings in all of its harmful components impairing mechanisms that protect the airways and lungs. Chronic respiratory health consequences are a complex multi-step silent process. By the time clinical manifestations require medical attention, several structural and functional changes have already occurred. The respiratory system has to undergo an iterative process of injury, healing and remodeling with every exposure. METHODS: To have a better understanding of the initial changes that take place when first exposed to environmental tobacco smoke, we have developed an exposure model, using the frog palate that closely represents the features of obstructive airways where ciliary dysfunction and mucus hypersecretion occur. RESULTS: Mucus transport was significantly reduced, even after exposure to the smoke of one cigarette (p < 0.05) and even further with 4-cigarettes exposure (p < 0.001). Morphometric and ultrastructural studies by SEM show extensive areas of tissue disruption. Gelatinase zymography shows activation of MMP9 in mucus from palates exposed to tobacco smoke. CONCLUSIONS: The clearance of mucus on the frog palate is significantly reduced after exposure to environmental tobacco smoke. Cilia and the extracellular matrix are anatomically disrupted. Tobacco smoke triggers an increased activity of matrix metalloproteinases associated with a substantial defoliation of ciliated epithelium. These studies enhance the knowledge of the changes in the mucociliary apparatus that occur initially after exposure to environmental tobacco smoke, with the goal of understanding how these changes relate to the genesis of chronic airway pathologies in humans. PMID- 15357882 TI - In vivo clearance of surfactant lipids during acute pulmonary inflammation. AB - BACKGROUND: A decrease in pulmonary surfactant has been suggested to contribute to the lung dysfunction associated with pulmonary inflammation. A number of studies have implicated surfactant clearance as a possible mechanism for altered pool sizes. The objective of the current study was to specifically investigate the mechanisms of surfactant clearance in a rodent model of acute pulmonary inflammation. METHODS: Inflammation was induced by intrapulmonary instillation of lipopolysaccharide (LPS: 100 microg/kg). Lipid clearance was assessed at 18 and 72 hours post-LPS instillation by intratracheal administration of radiolabel surfactant-like liposomes 2 hours prior to isolation and analysis of inflammatory cells and type II cells. RESULTS: At both 18 and 72 hours after LPS instillation there was significantly less radioactivity recovered in the lavage fluid compared to respective control groups (p < 0.05). At both time points, the number of cells recovered by lavage and their associated radioactivity was greater compared to control groups (p < 0.01). There was no difference in recovery of radioactivity by isolated type II cells or other cells obtained from enzymatic digestion of lung tissue. CONCLUSION: These results show that increased clearance of surfactant lipids in our model of acute pulmonary inflammation is primarily due to the inflammatory cells recruited to the airspace and not increased uptake by alveolar type II cells. PMID- 15357883 TI - A mechanism of airway injury in an epithelial model of mucociliary clearance. AB - We studied the action of sodium metabisulphite on mucociliary transport in a frog palate epithelial injury model, hypothesizing that it may be useful for the study of mechanisms of airway injury. Sodium metabisulphite (MB) releases SO2 on contact with water. SO2 is a pollutant in automobile fumes and may play a role in the exacerbation of airway disease symptoms. We first investigated its effect on mucociliary clearance. MB 10(-1) M, increased mucociliary clearance time (MCT) by 254.5 +/- 57.3% of control values, (p < 0.001, n = 7). MB 10(-4) and 10(-2) M did not interfere with mucus clearance time compared to control values. In MB-treated frog palates, MCT did not return to control values after one hour (control, 97.3 +/- 6.3% vs. MB, 140.9 +/- 46.3%, p < 0.001, n = 7). Scanning EM images of epithelial tissue were morphometrically analyzed and showed a 25 +/- 12% loss of ciliated cells in MB palates compared to controls with an intact ciliary blanket. Intact cells or groups of ciliated cells were found in scanning EM micrographs of mucus from MB-treated palates. This was associated with increased matrix metalloproteinase (MMP-9) activity in epithelial tissue and mucus. We suggest that the loss of ciliated cells as a result of MMP-9 activation prevented full recovery of MCT after MB 10(-1) M. The mechanism of action may be on epithelial cell-cell or cell-matrix attachments leading to cell loss and a disruption of MCT. Further studies are warranted to determine whether this is an inflammatory mediated response or the result of a direct action on epithelial cells and what role this mechanism may play in the progression to chronic airway diseases with impaired mucociliary clearance. PMID- 15357885 TI - Hearing aid use in nursing homes. Part 1: Prevalence rates of hearing impairment and hearing aid use. AB - Although hearing impairment is among the most common chronic ailments of older persons, the problem is even greater among those in nursing homes. It is estimated that 70% to 90% of elderly residents in long-term care facilities have some degree of hearing impairment. Despite this high prevalence, significant underuse of hearing aids or other assistive devices exists, especially among those with dementia. OBJECTIVES: The current study assessed rates of hearing impairment and hearing aid use among residents in a large, mid-Atlantic nursing home. SETTING AND DESIGN: In a cross-sectional survey, interviews were conducted at a large, mid-Atlantic nonprofit nursing home. PARTICIPANTS: Reports were obtained from 279 resident-caregiver dyads, as well as from the Minimum Data Set (MDS) and research assistants performing the structured interviews. MEASUREMENTS: Measures included the MDS and the Barriers to Hearing Aid Use Questionnaire, which was developed for this study. RESULTS: Results show an inconsistency in reports of both hearing impairment and hearing aid use by multiple sources, as well as the underuse of hearing aids. These inconsistencies suggest that gross hearing impairment is undetected in clinical practice in over half of the residents. CONCLUSION: The findings highlight the need for regular hearing assessments in the nursing home population. They also suggest the need for evaluation of institutional-level policies regarding screening and hearing aid management, as well as the need for further information regarding potential barriers to effective hearing aid use. These issues are addressed in part 2 of this study following this report. PMID- 15357886 TI - Hearing aid use in nursing homes. Part 2: Barriers to effective utilization of hearing AIDS. AB - OBJECTIVE: This study examined barriers to hearing aid use among persons who were reported to have a hearing aid and among those reported to have hearing difficulties but no hearing aids. SETTING: Interviews were conducted at a large, mid-Atlantic nonprofit nursing home. PARTICIPANTS: Both nursing home residents (279) and nursing staff members (51) were interviewed. DESIGN AND MEASUREMENTS: In a cross-sectional survey of nursing home residents, brief structured interviews were performed by trained research assistants with both residents and caregivers to obtain information regarding residents' hearing ability, hearing aid use and daily maintenance, and potential barriers to such use. RESULTS: Among residents reported to have hearing problems but who did not use a hearing aid, the major problem was neglect of the issue; participants did not know why residents did not have a hearing aid, residents had not had hearing evaluations, and staff members were not aware of hearing problems in residents. Among residents who did use a hearing aid, the majority (69%) of those for whom information was available had problems with the devices. The most common problems reported were that the device was hard or inconvenient to use, it did not fit well or hurt, and the device was not functioning well. The vast majority (86%) needed help taking care of the hearing aids. Close to half of the staff members had not received any training in the use or maintenance of the devices. Lack of delegation of responsibility for the management of hearing was identified for 29%, and relatives were used for maintenance of hearing aids in 14% of residents with hearing aids. CONCLUSIONS: Barriers to hearing aid use are therefore complex and multifactorial, involving lack of system commitment to utilization of hearing aids, lack of knowledge by staff members, inappropriate delegation and care procedures, hearing aid design and fit issues, and difficulties for residents in handling the hearing aids. Addressing these issues requires change on multiple levels, including change at the institutional level, concerning policy and training; change at the unit level, regarding care procedures and follow up; change at the individual level, providing better checks of fit and function of the hearing aids; and finally, change at the societal level, addressing design and cost issues for hearing aids in this population. PMID- 15357887 TI - Quality indicators for the management of medical conditions in nursing home residents. AB - PURPOSE: The purpose of this study was to develop a set of specific care processes associated with better outcomes for general medical conditions identified as quality improvement targets for institutionalized vulnerable elders. METHODS: A national panel of nursing home experts used a modified-Delphi process to rate the validity (process linked to improved outcomes) and feasibility (of implementation and measurement) of candidate measures for depression, diabetes, hearing impairment, heart failure, hypertension, ischemic heart disease, osteoarthritis, osteoporosis, pneumonia, stroke, and vision impairment. Each quality indicator was written as an "if" statement, describing persons to whom the quality indicator applied followed by a "then" statement identifying the care process to be provided. A separate clinical committee reviewed the resulting set of indicators. RESULTS: One hundred fourteen quality indicators were identified across the 11 medical conditions. The quality indicators capture a broad range of medical care addressing assessment, management, and follow up. Fifty-five indicators (48%) were identical to quality measures for community-dwelling vulnerable elders. A limited number were rated as questionably feasible to implement or measure (6 and 2, respectively). Thirty eight (33%) would not be applied to measures of care quality for persons with advanced dementia or poor prognosis. CONCLUSIONS: Explicit care processes linked to improved nursing home outcomes for general medical conditions can be identified. Most of these care processes can be measured by medical records or interview. Nursing home quality measures for medical conditions must account for exclusions related to poor prognosis and advanced dementia. PMID- 15357888 TI - Feasibility of quality indicators for the management of geriatric syndromes in nursing home residents. AB - PURPOSE: The assessment and management of dementia, falls and mobility disorders, malnutrition, end-of-life issues, pressure ulcers, and urinary incontinence have been identified as important quality improvement targets for vulnerable elders residing in nursing homes. This study aimed to identify valid and feasible measures of specific care processes associated with improved outcomes for these conditions. METHODS: Nine experts in nursing home (NH) care participated in a modified Delphi process to evaluate potential quality indicators (QIs) for care in NHs. Panelists met and discussed potential indicators before completing confidential ballots rating validity (process associated with improved outcomes), feasibility of measurement (with charts or interviews), feasibility of implementation (given staffing resources in average community NHs), and importance (expected benefit and prevalence in NHs). The NH panel's median votes were used to identify a final set of QIs that were subsequently reviewed by a clinical oversight committee. RESULTS: Sixty-eight geriatric syndrome QIs were identified as valid and important in NH populations. Panelists assessed 12 (18%) of these QIs as having questionable feasibility to implement in average community nursing homes trying to provide quality care. Nine (13%) would not be included in systems assessing quality of care for persons with advanced dementia or poor prognosis. CONCLUSIONS: Steps of care critical to the assessment and management of geriatric syndromes in NHs were identified. Feasibility is an important issue for a significant number of these, indicating that much remains to be done to design systems that efficiently and reliably implement these care processes. PMID- 15357889 TI - Analyses of nursing home residents with diabetes at admission. AB - OBJECTIVE: The objective of this study was to profile nursing home residents with diabetes at admission to the nursing facility. METHODS: We used all admission assessments in the Minimum Data Set recorded throughout the United States during 2002 to identify 144,969 residents with diabetes, or 26.4% of all admissions. RESULTS: Only approximately one fourth of residents with diabetes were projected to have stays in the facility of 90 days or less when admitted. Heart and circulatory comorbidities were common among residents with diabetes at admission, as was depression. More than half of residents with diabetes were in pain at admission. A majority of residents with diabetes were either totally dependent or required extensive assistance in the self-performance of many activities of daily living and more than one third were at least moderately impaired in cognitive performance. CONCLUSIONS: Residents with diabetes could be one of the most "heavy care" groups in nursing facilities, as demonstrated by their levels of functional disability and prevalence of serious comorbid conditions. The care provided to residents with diabetes should address depression, pain, and low rates of advance care planning. PMID- 15357890 TI - Evidence-based medicine: is it relevant to long-term care? AB - Internists and family physicians are trained in evidence-based medicine (EBM). The philosophy of incorporating clinical research "evidence" into medical practice is taught at the undergraduate and graduate levels of medical training. Practicing physicians incorporate EBM into their practices in variable degrees depending on their training, comfort level, and the nature of their practices. The evidence-based process has not clearly permeated the practice of long-term care, nor does it indisputably transfer from the office and hospital settings. This article discusses the applicability of EBM to the long-term care setting and some of the barriers to implementation. PMID- 15357891 TI - Current controversies in endocrinology: screening of asymptomatic elderly for subclinical hypothyroidism. AB - Thyroid disorders are common in elderly patients. In fact, the most common form of thyroid dysfunction in the elderly is subclinical hypothyroidism. Subclinical hypothyroidism is a biochemical state characterized by an increased serum thyroid stimulating hormone and normal levels of serum-free T4 and free T3. Much attention has been focused on this clinical entity recently, but it remains controversial whether early thyroid replacement therapy improves outcomes in elderly patients with asymptomatic subclinical hypothyroidism. There is a dearth of critical evidence on the effects of subclinical hypothyroidism on target tissues, the natural history of early thyroid dysfunction, and the net benefits and harms of long-term treatment with thyroid hormones. This article reviews the available information on the aging thyroid gland, subclinical hypothyroidism, and issues concerning treatment in asymptomatic elderly patients. PMID- 15357892 TI - Development and evaluation of a charge capture program for long-term care providers. AB - OBJECTIVE: Clinicians often have difficulty determining the appropriate Current Procedural Terminology Evaluation and Management code to assign to the type and intensity of patient care they provide. The purpose of this study was to develop, implement, and evaluate a handheld charge capture program for use by providers in the long-term care setting. DESIGN: Using a pre-post study design, we compared the coding accuracy and user satisfaction of an established paper process with a handheld charge capture program created for this study by means of: (1) preimplementation and postimplementation assessment of coding accuracy, and (2) preimplementation and postimplementation clinician survey. SETTING: We studied an academic division of geriatric medicine. PARTICIPANTS: Participants consisted of six clinicians who currently spend at least 50% of their clinical time practicing in the long-term care setting. INTERVENTION: A handheld charge capture program to replace the current paper-based charge capture process was reviewed. RESULTS: Overall coding accuracy improved by approximately 20% when the handheld program was used instead of a paper coding process. The majority of clinicians found that the handheld program was more widely available, efficient, easier to use, and encouraged the participants to document more completely and accurately in the patient's medical record. CONCLUSION: A handheld billing and coding program used by clinicians who provide care for long-term care residents is not only feasible, but leads to an improvement in coding accuracy when compared with a paper process. In addition, clinician satisfaction toward the billing and coding processes improved with the use of the handheld program. PMID- 15357893 TI - Effective pain management in the long-term care setting. AB - It is essential that physicians and midlevel practitioners who care for residents in long-term care facilities be proficient in the recognition, assessment, and treatment of chronic pain. A holistic approach to the physical, emotional, social, and spiritual components of a resident's total pain and distress must be integrated into the palliative aspects of long-term care medicine. Furthermore, all practitioners must recognize and effectively manage, prevent, and/or minimize the occurrence of acute pain, breakthrough pain, incidental pain, and disturbance pain that frequently are superimposed on a resident's chronic pain. PMID- 15357894 TI - Analyses of nursing home residents with diabetes at admission. PMID- 15357895 TI - Year in review: National Pressure Ulcer Long-Term Care Study (NPULS). PMID- 15357897 TI - Comprehensive geriatric assessment training. PMID- 15357899 TI - Adenoviral vectors for gene replacement therapy. AB - Adenovirus-based vectors are promising vehicles for gene replacement therapy due to their ability to efficiently transduce a wide variety of proliferating and non proliferating cells. Over the past decade, different versions of adenoviral vectors (Ads) have been developed. These vectors can be classified into two major categories, based on whether the viral coding sequences are partially (first or second-generation Ads) or completely deleted (helper-dependent or gutted Ads). Both types of Ads have been tested in a variety of gene delivery studies, and major obstacles to their clinical application have been identified. Currently, innate and adaptive host immune responses to Ads remain major challenges, limiting both the initial viral dose and the effectiveness of subsequent administrations. Recent developments in vector design and delivery methods have improved the potential of Ads for successful gene therapy application. PMID- 15357900 TI - Prion diseases and the spleen. AB - Transmissible spongiform encephalopathies are fatal neurodegenerative disorders that include Creutzfeldt-Jakob disease in humans, bovine spongiform encephalopathy and scrapie in sheep and goats. Transmissible spongiform encephalopathies are thought by some to result from changes in the conformation of a membrane glycoprotein called PrPC (prion protein) into a pathogenic form, PrPSc, which constitutes the major component of an unprecedented type of infectious particle supposedly devoid of nucleic acid. Although there is no primary immunological response to the infectious agent, several lines of evidence indicate an involvement of the lymphoreticular system in the development of prion diseases. Studies in rodents have shown that after peripheral infection, uptake of the scrapie agent is followed by an initial phase of replication in the lymphoreticular system, particularly the spleen and lymph nodes. Moreover, infectivity titers in lymphoreticular organs reach a maximum relatively quickly, well before those in the brain, and then maintain a plateau for the remainder of the disease progression. The presence of PrPSc in peripheral lymphoid organs of all cases of variant Creutzfeldt-Jakob disease strongly underscores the importance of the lymphoreticular system. Thus, a better understanding of the cells participating in PrPSc replication and dissemination into the central nervous system is of particular interest. This review will therefore discuss the present knowledge of the role of the spleen in transmissible spongiform encephalopathies as well as the participation of the different spleen cell types in the disease process. PMID- 15357901 TI - Influenza pandemics: can we prepare for the unpredictable? AB - Although no viruses are better understood or more intensively studied than the viruses of influenza, if the next influenza pandemic occurs within the next 5-10 years its control will depend on innovations in vaccine production developed more than 40 years ago, but not yet applied to the full extent demanded by our present hard-won knowledge of the epidemiology of the disease. We have become so enamored of the brilliant advances made in the interim in understanding the molecular biology of both virus and host that common sense and inexpensive implementation of proven and older methods of control have been neglected as an interim barricade. In this review, I have advocated a return to first principles, while embracing the promise and returns of contemporary research. With the assumption that the next pandemic virus will contain one of the 13 influenza A virus hemagglutinin subtypes not currently causing epidemic human disease, high-yield reassortant viruses of each of these subtypes should be produced with all dispatch and, in collaboration with industry, tested for production stability and immunogenicity in humans. From this archive, an appropriate reassortant could be selected within days or weeks, and production could ensue. If not a perfect match with the imminent pandemic virus, this "barricade vaccine" could stand as a first line of defense until supplanted by a definitive "rampart vaccine," matching better the emergent, potentially pandemic virus. PMID- 15357902 TI - Coxsackievirus-induced pancreatitis. AB - In humans, infections with the group B coxsackieviruses (CVBs) range from asymptomatic infections to chronic, debilitating diseases. The CVBs are associated with chronic inflammatory diseases of the pancreas, heart, and central nervous system. A major focus in CVB pathogenesis is to understand the mechanisms by which these viruses cause acute diseases that resolve or acute diseases that progress to chronic diseases. The present review explores CVB infections in the development of acute and chronic pancreatitis. Mouse models of CVB-induced pancreatitis share many features with the human diseases and are providing insight into the multi-faceted processes of pancreatic tissue repair and irreversible tissue destruction. The development and progression of CVB-induced pancreatic inflammatory disease is an extremely complex process, involving both viral and host factors. The review examines the roles of the virus and host in contributing to the disease process. Recent studies of global gene expression during CVB-induced pancreatitis have increased our understanding of host factors that influence the outcome of infection and have highlighted interrelationships among complex biological programs. As we unravel the complexity of the disease process, the information gained will lead to the design of therapeutics that not only prevent the progression of chronic inflammatory disease, but that also restore functionality of affected tissues and organs. PMID- 15357903 TI - Respiratory syncytial virus-induced immunoprotection and immunopathology. AB - Respiratory syncytial virus (RSV), a member of the Paramyxoviridae family, is a major clinical problem causing yearly epidemics of severe lower airway disease in both infants and the elderly. Attempts at vaccination have been frustrated by both the poor immunogenicity of this virus, and the severe immunopathology observed in early vaccine trials. Primary infection generally occurs in infancy, with approximately 5% of infected infants requiring hospitalization. Equally problematic is the apparent link between severe RSV disease and the later development of allergy and asthma. While there is no evidence that natural infection promotes Th2 predominance, development of enhanced eosinophilic disease in children receiving inactivated virus administered with a commonly used adjuvant demonstrated how easily the balance between immune-mediated protection and immune-mediated pathology can be perturbed. In this review we have focused on studies carried out in the mouse model aimed at determining the correlates of RSV protection and explaining the mechanism of vaccine enhanced immunopathology. PMID- 15357904 TI - High-risk papillomavirus infection is associated with altered antibody responses in genital tract: non-specific responses in HPV infection. AB - In order to gain more information about local humoral immune responses to HPV infection, we quantified IgG, IgM, secretory-IgA (S-IgA), and total-IgA by ELISA, and lysozyme and lactoferrin by TR-IFMA, in cervical and cervicovaginal secretions of 40 healthy women and 28 high-risk HPV infected patients (11 were HPV16+). IgG, total-IgA, and S-IgA concentrations in cervicovaginal secretions (p < 0.0001) and high IgG and total-IgA concentrations (p < 0.001 and p < 0.01, respectively) in endocervical secretions were significantly higher in HPV+ patients than in the healthy group. Since the S-IgA/total-IgA ratio was significantly lower in cervicovaginal (7.5%) and endocervical secretions (36.5%) in HPV+ women compared to the control group (p < 0.003 and p < 0.001, respectively), HPV could be responsible for an increase in local production of non-secretory IgA (monomeric and dimeric forms). IgG and total-IgA concentrations in cervicovaginal and endocervical secretions fell in the same general percentage range in both HPV16+ and HPV+ groups (80% and 15%, respectively). However, the S IgA/total-IgA ratio was much lower in HPV16+ than in HPV+ women, in both cervicovaginal secretions (3.4%) (p < 0.003) and in endocervical secretions (23.3%) (p < 0.001). Innate immunity proteins and local S-IgA response could not stop the spread of HPV infection in spite of high lysozyme and lactoferrin concentrations. HPV16+ disturbed the local humoral immune system, which could partly explain its low clearance. PMID- 15357905 TI - Depletion of peripheral blood T lymphocytes and NK cells during the course of ebola hemorrhagic Fever in cynomolgus macaques. AB - During the course of an experimentally induced Ebola virus (EBOVA) infection of cynomolgus macaques, peripheral blood mononuclear cells were isolated and characterized by multi-color flow cytometry. Both CD4+ and CD8+ lymphocyte counts decreased 60-70% during the first 4 days after infection. Among CD8+ lymphocytes, this decline was greatest among the CD8(lo) population, which was composed mostly of CD3- CD16+ NK cells. In contrast, the number of CD20+ B lymphocytes in the blood did not significantly change during the course of the infection. Phenotypic analysis of T lymphocyte subsets by flow cytometry failed to show evidence of a robust immune response to the infection. Apoptosis could be detected as early as day 2 postinfection among the CD8+ and CD16+ subsets of lymphocytes. Increased expression of CD95 (Fas) suggests that apoptosis may be induced via signaling through the Fas/Fas-L cascade. In contrast, the number of HLA-DR+ cells increased tenfold in the blood during the course of infection. These data suggest that EBOV may block dendritic cell maturation after infection, thereby inhibiting activation of lymphocytes and eliminating those subsets that are most likely to be capable of mounting an effective response to the virus. PMID- 15357906 TI - Cytotoxic T cell epitope in cattle from the attachment glycoproteins of rinderpest and peste des petits ruminants viruses. AB - The surface glycoproteins of rinderpest virus (RPV) confer protective immunity in cattle. We demonstrated that cattle immunized with a recombinant extracellular baculovirus expressing the hemagglutinin (H) protein of RPV (rECV-H) generate virus neutralizing antibody responses, bovine leukocyte antigen (BoLA) class II restricted helper T cell responses and BoLA class I restricted cytotoxic T cell (CTL) responses against RPV-H and hemagglutinin-neuraminidase (HN) glycoprotein of closely related Peste des petits ruminants virus (PPRV). In this study, employing autologous skin fibroblasts transiently expressing truncations of H and HN in a BoLA class I restricted lymphoproliferation assay, we have mapped a highly homologous domain (amino acids 400-423) on these proteins harboring a CTL epitope. Subsequently, based on sequence comparison with available BoLA class I binding motifs, we have identified a BoLA-A11 binding motif (amino acids 408-416) in the stimulatory domain. Autologous cells pulsed with a synthetic peptide corresponding to this sequence stimulated CTLs from rECV-H immunized as well as tissue culture attenuated RPV vaccinated cattle of different breeds and parentage. This is the first epitope identified in cattle on the attachment glycoproteins of RPV and PPRV. PMID- 15357907 TI - Expression of IL-12 receptor by neurons. AB - Interleukin (IL)-12, a key cytokine bridging innate and acquired immunity, is efficacious in enhancing recovery from experimental vesicular stomatitis virus (VSV) infection of the mouse central nervous system (CNS). This response is associated with the upregulation of neuronal nitric oxide synthase (NOS-1), independent of IFN-gamma and TNF-alpha. We hypothesized that neurons may respond directly IL-12. Our data are consistent with the expression of a functional IL-12 receptor (IL-12R) by neurons in culture and this receptor-ligand interaction results in the induction of an innate antiviral immune response. N18 cells, which did not express IL-12Rbeta2 were transfected with the IL-12Rbeta2 receptor gene; Koch's postulates were fulfilled, as clones derived from this transfection were reconstituted for IL-12 responsiveness. PMID- 15357908 TI - Induction of broad-based immune response against HIV-1 subtype C gag DNA vaccine in mice. AB - Prevention of HIV infection through an effective vaccine is need of the hour as per the AIDS pandemic scene, particularly in the developing world. Here we report the work done with gag gene construct pJWgagprotease49587 from HIV-1subtype C Indian strain. The construct pJWgagprotease49587 was tested positive for expression in COS-7 cells by p24 antigen capture ELISA, immunoblotting and by transmission electron microscopy that revealed virus like particle formation. Immunogenicity studies showed induction of good lymphoproliferative and cytotoxic (CTL) responses in Balb/c mice. The cytokine repertoire elicited showed a TH1 type of immune response. In an epitope mapping study, IFNgamma secretion by spleen cells from immunized mice was observed to seven peptides from different regions of gag. Recognition of multiple epitopes demonstrates elicitation of a broad based immune response against gag following immunization with the construct. In view of the high propensity of escape mutant induction during the course of HIV infection, it is encouraging to use immunogens eliciting viable immune responses to a broad spectrum of epitopes. Hence the construct pJWgagprotease49587 is a good candidate for immunogenecity testing in nonhuman primates as a probable vaccine candidate. PMID- 15357909 TI - Humoral immunity and the evolution of HIV-2. AB - Human immunodeficiency virus type 2 (HIV-2) evolved from the zoonotic transmission of simian immunodeficiency virus (SIV) that naturally infects sooty mangabeys found in West Africa. Using sera from HIV-2-infected humans, we discovered that an hypervariable region (the V4 loop) of HIV-2 induces antibody responses only weakly reactive against itself but strongly reactive against analogous sequences from the V4 loop of strains of SIV. Available sequence data indicates that all strains of HIV-2 have large deletions in the V4 region that truncate an immunodominant neutralizing B cell epitope among strains of SIV. Infection of a macaque with a sequenced clone of HIV-2 similarly elicited antibodies that poorly recognized the V4 loop of HIV-2 but readily bound to analogous SIV sequences. Our data are consistent with a scenario whereby a disparate antibody response directed against the V4 loop may have influenced the selective expansion and survival of HIV-2 in humans. PMID- 15357910 TI - Mutation of the "a" determinant of HBsAg with discordant HBsAg diagnostic kits. AB - Sera from 1003 in- and out-patients were investigated for hepatitis B surface antigen (HBsAg) mutation at Siriraj Hospital, Bangkok, Thailand. Individual samples were screened using two commercial HBsAg kits on automatic machine-based assays set up in parallel. The first kit was a sandwich ELISA kit that used monoclonal capture/monoclonal conjugate and color detection whereas the second was a sandwich MEIA, using monoclonal capture/polyclonal indicator and fluorochrome determination. Six specimens were found discordant by negative EIA and positive MEIA; two specimens of which were detectable of HBV DNA. Three out of four discordant results were confirmed by an anti-HBs neutralization assay. Based on direct sequencing, one HBsAg/anti-HBs sample with multiple mutations in the S gene was found. The mutation included the common glycine to arginine escape mutation at amino acid position 145 of the "a" determinant. The observation should alert the blood bank to the necessity of using screening kits capable of detecting HBV mutant carriers as well as providing verification for the phenomenon of vaccine-escape mutation in Thailand. PMID- 15357913 TI - V-1 immunitor. PMID- 15357911 TI - CMV antigen-specific CD4+ and CD8+ T cell IFNgamma expression and proliferation responses in healthy CMV-seropositive individuals. AB - CMV-specific CD4+ and CD8+ T cell IFNgamma expression and proliferation were measured in healthy volunteers by flow cytometry after CMV lysate or CMV pp65 or IE peptide pool stimulation. Cutoff values were set to maximize specificity (i.e., no false positive CMV-seronegatives). Sensitivity (defined as a positive response in CMV-seropositives to at least one of the 3 antigen preparations used) was 100% for CMV-specific CD4+ and CD8+ T cell IFN expression and CD4+ T cell proliferation and 95.4% for CMV-specific CD8+ T cell proliferation. All 22 CMV seropositive individuals had positive responses by at least three of these four measurements. These findings support the concept that a multiplicity of antigen specific functional immune responses and persistence of robust virus-specific CD4+T cells are important components of protective immunity in this chronic viral infection. PMID- 15357914 TI - An interview with Steven M. Watkins, Ph.D., president and chief scientific officer, Lipomics Technologies, Inc. Interview by Vicki Glaser. PMID- 15357915 TI - Development of an assay using 4-trifluoromethylumbelliferyl as a marker substrate for assessment of drug-drug interactions to multiple isoforms of UDP glucuronosyltransferases. AB - Glucuronidation is catalyzed by UDP-glucuronosyltransferases (UGTs) and is one of the most important pathways for elimination of xenobiotics. The aim of the present study was to develop an in vitro assay for assessment of drug-drug interactions related to UGTs applicable to early drug discovery. 4 Trifluoromethylumbelliferyl was tested as a marker substrate for six human recombinant expressed UGT isoforms: 1A1, 1A3, 1A4, 1A6, 1A9, and 2B7. It was shown that 4-trifluoromethylumbelliferyl was glucuronidated by all UGTs tested, except UGT1A4. By using a short HPLC gradient (7 min) and fluorescence detection, the enzyme kinetic parameters for these reactions were obtained. All reactions were found to follow classical Michaelis-Menten kinetics, with K(m) values between 29 microM (UGT1A9) and 80 microM (UGT1A3). The method was validated by using several known competitive inhibitors of UGTs. The most potent inhibition was observed for the reaction between 17alpha-ethynylestradiol and UGT1A1 (K(i) = 10.5 microM), and the weakest interaction was detected for acetaminophen and UGT1A9 (IC(50) > 1 mM). Taken together, we report the development of an assay using 4-trifluoromethylumbelliferyl as a marker substrate for five different human UGT isoforms suitable for the assessment of drug-drug interactions related to UGTs during early drug discovery. PMID- 15357916 TI - 96-well plate assay for sublethal metabolic activity. AB - A 96-well plate assay has been devised for estimation of sublethal metabolic activity for compounds administered to in vitro cell cultures during 6- and 24-h exposures. This screen combines a resazurin reduction assay with lactate production and glucose consumption rate assays to assess effects of compounds on both culture viability and metabolic inhibition. In this article, the assay is demonstrated by determining the extent to which five glycolysis inhibitors, namely, phloretin, 2-deoxyglucose, iodoacetate, fluoride, and oxamate, induce metabolic inhibition without cell death for in vitro fibroblast cell cultures. During 6-h exposures, iodoacetate was found to be the most potent inhibitor of glycolysis, whereas iodoacetate and phloretin were most cytolethal. 2 Deoxyglucose had the largest sublethal metabolic range, spanning just over two orders of magnitude in concentrations between its IC(50) values for cytolethality and metabolic inhibition. This method provides a simple and inexpensive way to determine global metabolic and cytolethal effects of compounds for in vitro cell culture systems. It should be of use directly for large-scale screening and ranking of compounds during drug discovery and development, in conjunction with or following some more direct measure of therapeutic efficacy of a prospective drug candidate. Moreover, firsthand determination of the concentrations over which a compound has lethal, sublethal metabolic, or no such effects should serve as a cost-effective and time-saving first step in a given research study, preceding more expensive, lengthy, and/or detailed in vitro methods. PMID- 15357917 TI - Application of real-time cell electronic sensing (RT-CES) technology to cell based assays. AB - Label-free detection emerges as a new approach in the development of technologies for cell-based screening assays. Unlike the classic detection methods that use fluorescence, radioisotope, luminescence, or light absorption, label-free detection directly measures the cell function without using a labeled molecule. The advantages of label-free detection include a simple homogeneous assay format, noninvasive measurement, less interference with normal cell function, kinetic measurement, and reduced time for assay development. Here, we have applied the electrical impedance detection method in a real-time cell electronic sensing (RT CES trade mark ) system for cell-based assays. The cell growth rate measured by this RT-CES system was comparable to actual cell number counted manually. In addition, cell proliferation, cytotoxicity, cytoprotection, cell growth inhibition, and apoptosis data generated by this RT-CES system correlated with those determined by the classic methods. The conclusion is that the RT-CES system is a useful tool for label-free detection of certain cell-based parameters. PMID- 15357918 TI - High throughput screening via mass spectrometry: a case study using acetylcholinesterase. AB - Mass spectrometry-based screening can be applied to a wide range of targets, including those intractable targets that use substrates such as lipids, fatty acids, phospholipids, steroids, prostaglandins, and other compounds not generally amenable to conventional screening techniques. The major limitation to this approach is throughput, making HTS via mass spectrometry impractical. We present a mass spectrometry-based technique and hardware for lead discovery applications. Mass spectrometry enables the design of label-free assays using biologically native substrates for a wide range of enzymatic targets. This system can be used for the direct quantification of analytes in complex reaction mixtures with typical throughputs of 4-5 s per sample. A mass spectrometry-based assay was developed to identify inhibitors of acetylcholinesterase, an enzyme with clinical importance in Alzheimer's disease. The system was used to screen a small chemical library. Several potent inhibitors were identified, and the IC(50) values of the inhibitors were determined. PMID- 15357919 TI - Expression of amyloid-beta 1-40 and 1-42 peptides in Capsicum annum var. angulosum for oral immunization. AB - Alzheimer's disease (AD), the leading cause of dementia in the elderly population, still remains without an effective treatment. The accumulation and deposition of the amyloid-beta peptide (Abeta) in the brain is thought to be a key event in the pathogenesis of AD. Recently, a novel exciting technology has been investigated to combat AD: new immunotherapeutic approaches have been described that are based on vaccination with the Abeta peptide itself, and this has been shown to induce functionally beneficial anti-Abeta antibody responses in different transgenic animal models of AD. Here we report the high level expression of GFP-Abeta1-40 and 1-42 peptides in Capsicum annum var. angulosum (green pepper) using a new tomato mosaic tobamovirus-based hybrid replication vector. After preinoculation of Nicotiana benthamiana plants with the in vitro transcript of the vector, the isolated virions were used to inoculate green pepper, which accumulated the GFPAbeta1-40 or 1-42 fusion proteins to a level of 100 microg/g of leaves 7 days after inoculation. These results make it possible to test whether oral immunization by feeding plant samples could stimulate antibody production against Abeta peptides. PMID- 15357920 TI - Micro parallel liquid chromatography: enabling technology for discovery analytical chemistry. AB - Since the introduction of combinatorial chemistry, compound libraries have undergone a significant increase in size and diversity. The ensuing expansion and diversification of compound libraries have resulted in increased demand for analytical throughput. Following the evolution of new technologies for generating lead compounds and targets and the desire to increase research and development productivity, analytical chemistry is now gaining attention as a bottleneck that would benefit from advances in instrumentation for increased analytical throughput. The commercial introduction of the Veloce trade mark micro parallel liquid chromatography system from Nanostream offers discovery analytical chemists the capability to analyze 24 samples in parallel with as little as 0.5 microl of sample. The system offers a scalable analytical approach to address bottlenecks in historically underserved areas, such as compound library purity screening, as well as higher value-added applications, such as log P determination and aqueous solubility assessment. This article describes the Veloce system and presents representative data from several discovery analytical applications. PMID- 15357921 TI - Single-molecule techniques for drug discovery. AB - Single-molecule techniques offer a number of key benefits over conventional in vitro assay methods for drug screening, as they use less material and unlock the ability to observe transient states. By observing such states, it should be possible to screen for chemical compounds that isolate these steps. The benefit of this is twofold: (a) inhibitors can be found that target key phases in biochemical processes, e.g., transcription initiation; and (b) the total number of drug targets increases as many biochemical processes consist of many transient steps, e.g., transcription promoter binding, initiation, elongation, and termination. Although single-molecule methods offer exciting opportunities for new ways of discovering drugs, there are a number of obstacles to their adoption for drug screening. The main hurdle is to develop robust apparatus that will allow many thousands of individual single molecule experiments to be performed in parallel. By using recently developed integrated microfluidics technology, this hurdle may be overcome. Here, a number of potential single-molecule approaches to drug screening are presented along with a discussion of the benefits and technical obstacles that must be overcome. PMID- 15357922 TI - Optimizing the hit-to-lead process using SPR analysis. AB - Secondary screening and lead optimization, where a large number of "hit" compounds are refined to a viable set of "lead" drug candidates, are considered to be bottlenecks to the drug discovery process and are targets for streamlining. Surface plasmon resonance (SPR) is a nonlabel technology that can generate kinetic data on biomolecular interactions. This allows researchers to quantitate the binding characteristics of lead compounds with their targets in terms of affinity, specificity, and association/dissociation rates in parallel. The latest generation of SPR biosensors integrate the hit-to-lead process and generate a greater depth of information, providing answers that cannot be addressed by traditional end-point assays. This allows users to make more informed choices on the selection of candidate molecules prior to preclinical development. A number of studies have used SPR biosensors in secondary screening, lead optimization, quantitative structure-activity relationship analysis, and predictive adsorption, distribution, metabolism, excretion, and/or toxicity evaluations. PMID- 15357923 TI - Reference systems for kinase drug discovery: chemical genetic approaches to cell based assays. AB - Protein kinases play key roles in a number of diseases, including cancer, inflammation, and diabetes. Disregulation of kinase-based signal transduction networks results in aberrant cell differentiation, activation, proliferation, and invasion. The growing importance of kinases as a major class of drug targets across multiple large clinical indications, together with the large number of kinases in the genome (~518), has generated a critical need for technologies that enable the identification of potent and selective kinase inhibitors with good drug-like properties. In this review, we describe methods used for developing cell-based assays for kinase inhibitors, discuss advantages and disadvantages of each approach, and describe new chemical genetic methods as reference systems for establishing cell-based assays and their use for functional selectivity profiling of kinase inhibitors. PMID- 15357924 TI - Rapid structure-activity and selectivity analysis of kinase inhibitors by BioMAP analysis in complex human primary cell-based models. AB - Rapid, quantitative methods for characterizing the biological activities of kinase inhibitors in complex human cell systems could allow the biological consequences of differential target selectivity to be monitored early in development, improving the selection of drug candidates. We have previously shown that Biologically Multiplexed Activity Profiling (BioMAP) permits rapid characterization of drug function based on statistical analysis of protein expression data sets from complex primary human cellbased models of disease biology. Here, using four such model systems containing primary human endothelial cells and peripheral blood mononuclear cells in which multiple signaling pathways relevant to inflammation and immune responses are simultaneously activated, we demonstrate that BioMAP analysis can detect and distinguish a wide range of inhibitors directed against different kinase targets. Using a panel of p38 mitogen-activated protein kinase antagonists as a test set, we show further that related compounds can be distinguished by unique features of the biological responses they induce in complex systems, and can be classified according to their induction of shared (on-target) and secondary activities. Statistical comparisons of quantitative BioMAP profiles and analysis of profile features allow correlation of induced biological effects with chemical structure and mapping of biological responses to chemical series or substituents on a common scaffold. Integration of automated BioMAP analysis for prioritization of hits and for structure-activity relationship studies may improve and accelerate the design and selection of optimal therapeutic candidates. PMID- 15357932 TI - Highly active antiretroviral therapy in Brazil: the challenge of universal access in a context of social inequality. AB - OBJECTIVE: To investigate trends in AIDS mortality and incidence in Brazil over the period of 1984 to 2000 and to assess the impact of the introduction of universal access to highly active antiretroviral therapy (HAART) in the country in 1996. METHODS: Data from the Brazilian disease notification system and the national mortality information system were used to calculate annual region specific and sex-specific AIDS incidence and mortality rates. We also calculated sex- and region-specific ratios of the number of AIDS deaths in one year to the number of AIDS cases notified two years earlier. RESULTS: AIDS mortality rates for both men and women and in all five of the geographic regions of Brazil declined following introduction of HAART, despite continued growth in AIDS incidence. The ratio of the number of AIDS deaths in one year to the number of AIDS cases notified two years earlier for men equalized rapidly with the ratio for women following introduction of HAART. More recently, AIDS incidence declined for both sexes and in most of the regions of Brazil. CONCLUSIONS: Despite Brazil's resource limitations and disparities in wealth between men and women and among the country's regions, the introduction of universal access to HAART in Brazil has helped achieve impressive declines in AIDS mortality, and it may also be contributing to declines in AIDS incidence. PMID- 15357933 TI - [Evaluation of an educational module on dengue and Aedes aegypti for schoolchildren in Honduras]. AB - OBJECTIVE: To evaluate the effectiveness of a special course on environmental health and dengue, given to primary school students and intended to promote behavioral changes in the mothers of those students that would lead to the safe handling of water, adequate disposal of trash, and control of household breeding sites of Aedes aegypti, the mosquito vector for dengue. METHODS: In the period from April to November 2002 a community field trial was carried out that included four public schools in the city of Comayaguela, Honduras. Two of the schools formed the intervention group, and the two others made up the comparison or control group. The data collection activities before and after the intervention included knowledge tests for the schoolchildren and the teachers; surveys of the knowledge, attitudes, and practices of the students' mothers; entomological surveys (systematic checking in order to find water containers, to search for mosquito larvae) in the homes of the schoolchildren; in-depth interviews with the students' mothers; observation in the classrooms; and postintervention focus group with the teachers. RESULTS: During the intervention period the largest epidemic of classical dengue ever in the history of Tegucigalpa, the capital of Honduras, occurred. That event prompted the Secretariat of Health to undertake large-scale control activities. In the two intervention schools there was a significant increase in the students' knowledge of the following three variables: dengue caused by a virus, life cycle of the vector, and reduction of breeding sites, which is the most effective measure for controlling the vector (P < 0.0001 for all those increases in knowledge). There were also significant increases in the knowledge of the same variables among the teachers in the intervention schools: dengue caused by a virus (P = 0.03), life cycle of the vector (P = 0.01), and reduction of breeding sites (P = 0.004). Reducing the number of breeding sites as being the best measure for controlling Aedes aegypti was the knowledge variable that showed the greatest change among the mothers in the intervention group (P = 0.02). The values for two of the entomological indices (the House index and the Breteau index) were better in the intervention group than in the control group, but the difference was not statistically significant. However, there was a statistically significant difference in the Breteau index values between the two control group schools and the one intervention school where the education course was implemented more adequately than in the other intervention school. CONCLUSIONS: The application of educational interventions directed to schoolchildren, as a part of a comprehensive plan for controlling Aedes aegypti, is an effective way to raise the awareness of teachers and parents with respect to the problem of dengue, and it also helps in encouraging family members to be involved in reducing breeding sites in the home. In moving to control the dengue vector, this strategy should take into consideration the involvement not only of families but also of civic and public institutions in the community. Given our results, it appears that this approach of using a special course for schoolchildren can constitute a fundamental pillar in the strategy for integrated dengue control. PMID- 15357934 TI - [Risk factors for sexually transmitted diseases in prostitutes and transvestites in Ribeirao Preto (SP), Brazil]. AB - OBJECTIVE: To describe the demographic and socioeconomic characteristics of sex workers in the city of Ribeirao Preto, which is in the state of Sao Paulo, Brazil, and to investigate the risk factors for sexually transmitted diseases (STDs) in this population. METHODS: All the areas in the city where female or male prostitutes or transvestites work or live were visited. The participants answered a questionnaire that collected sociodemographic data and information concerning risk factors for STDs. A social worker who for more than five years had carried out educational activities with sex workers in Ribeirao Preto was responsible for the interviews. RESULTS: The 449 female prostitutes, 13 male prostitutes, and 53 male transvestite sex workers who were included in the study make up a young population, with little schooling and a low socioeconomic level. A majority of them were born in states other than Sao Paulo, and a third of them still maintain a residence in some other state. In comparison to the prostitutes (the female and male prostitutes considered together), the transvestites had a significantly higher risk for STDs, which was reflected in their number of years as sex workers, average number of sexual partners per day, history of ulcerative STDs, practice of anal sex, use of illegal noninjectable drugs (especially crack), and history of incarceration. Exposure to alcohol was the only risk factor found more frequently in the prostitutes. Using a condom with a steady partner was less frequent than was condom use in commercial sex, for both the prostitutes and the transvestites. CONCLUSIONS: Sex workers in Ribeirao Preto, especially transvestites, are socially marginalized and at high risk for STDs. Public health services should focus more attention on this population by developing prevention programs and by supporting additional research that could provide more detailed knowledge concerning the specific risk factors that put this population at risk for STDs. PMID- 15357935 TI - [Mortality from suicides: Mexico, 1990-2001]. AB - OBJECTIVE: To describe mortality from suicides in Mexico in 2001, as well as the main changes in the methods used to commit suicide and in trends by age and gender that have been observed since 1990, both for the country as a whole and for each state. METHODS: For this descriptive study we utilized as information sources the official mortality records of the National Institute of Statistics, Geography, and Informatics (Instituto Nacional de Estadistica, Geografia e Informatica) for the period of 1990 through 2001. To calculate mortality rates we used the populations estimated in 2002 by the National Population Council (Consejo Nacional de Poblacion). Mortality was described by sex, age group, and state, along with the changes seen over the period of 1990 through 2001 in the rates and methods of suicide. We used the direct method to standardize the rates, using as a reference the population data for the year 2000. To make statistical comparisons of the trends by age group and gender we used a test of parallelism utilizing the F statistic. The level of statistical significance of differences in suicide methods was determined with the chi-square test. RESULTS: During 2001, 3,784 suicides were registered (3,110 of them in men and 674 in women), which represents a rate of 3.72 deaths from suicide per 100,000 persons, in a total national population of 101.8 million inhabitants. The states with the highest suicide mortality were Campeche and Tabasco (9.68 and 8.47 per 100,000, respectively). The lowest rates were seen in Chiapas and the state of Mexico (1.03 and 1.99 per 100,000, respectively). In 2001, mortality from suicides per 100,000 persons was 6.14 in men and 1.32 in women. The greatest increase by age group was seen in women 11-19 years old (from 0.8 per 100,000 in 1990 to 2.27 per 100,000 in 2001). The largest increase in men also occurred among those 11-19 years old (from 2.6 per 100,000 in 1990 to 4.5 per 100,000 in 2001). The highest rate (13.62 per 100,000 persons) was seen in men over 65 years of age. There were changes in suicide methods, with hanging now being the method most frequently used by both men and women. CONCLUSIONS: Suicide is a growing problem in Mexico, and controlling it requires immediate steps. The rapid increase in this phenomenon, particularly among young men and young women, should produce a call to action to quickly implement measures aimed at reducing the magnitude of suicide. PMID- 15357936 TI - Gender inequalities in health among older Brazilian adults. AB - OBJECTIVE: To investigate gender differences among older Brazilians in their health status and their use of health services. METHODS: Participants were individuals aged 60 years and older included in a national household survey conducted in Brazil in 1998. Data were analyzed by multiple logistic regression, taking into account the design effect due to multistage sampling. RESULTS: There were differences in the health and living conditions of older men and older women that were not explained by age or place of residence. Older women had worse indicators of schooling and personal income but better indicators of housing standards and per capita household income. The older women also reported more chronic diseases, had poorer indicators of independence and physical mobility, sought health services more often, and reported more medical visits in the previous year. Despite their apparent worse health conditions, elderly women in urban areas had lower hospitalization rates in the previous year (odds ratio = 0.89; 95% confidence interval, 0.82-0.96) than did elderly men in urban areas. CONCLUSIONS: Our results indicate that among older Brazilians there are gender inequalities in health that cannot be explained by age and place of residence. The findings raise questions on how health, socioeconomic, and cultural factors influence gender patterns of seeking and using health care in later life in the country. As pressures on health care and health funding increase in Brazil as a result of the aging of the population, there is a need to take a gender perspective into account. PMID- 15357937 TI - [Using surveys of schoolchildren to evaluate coverage with and opportunity for vaccination in Costa Rica]. AB - OBJECTIVE: To identify differences in the level of coverage of and opportunity for vaccination among schoolchildren in three areas in Costa Rica with different characteristics: an urban area (with the highest level of socioeconomic development of the three areas), a rural area (with a medium level of socioeconomic development), and a border area (a rural area in northern Costa Rica, on the border with Nicaragua, with the lowest level of socioeconomic development and the highest proportion of foreign immigrants). METHODOLOGY: Following selection of schools by proportional probability, surveys were used with children chosen at random from the first and second grades of elementary schools in the three areas: urban (961 students), rural (544 students), and border (811 students). The data on the vaccines that had been administered were obtained from the children's vaccination cards. Differences among the three areas were evaluated: (1) in the coverage with BCG; with three doses of diphtheria tetanuspertussis vaccine (DTP3); with three doses of oral polio vaccine (OPV3); with the first dose of measles-mumps-rubella vaccine (MMR1); and with the second dose of MMR vaccine (MMR2) and (2) in the "opportunity" for the children having received DTP1 + OPV1 before 3 months of age, DTP3 + OPV3 before 7 months of age, and DTP4 + OPV4 + MMR1 before 24 months of age. RESULTS: Out of all the students who had been selected, 80% of them in the urban area had a vaccination card, 73% did in the rural area, and 72% did in the border area (P < 0.05). The coverage levels for BCG, DTP3, and OPV3 were each over 95% in both the urban area and the rural area; however, the coverage levels were significantly lower (P < 0.05) in the border area: BCG, 83%; OPV3, 88%; and DTP3, 88%. Coverage with MMR1 and MMR2 was similar in the three areas. The percentage of schoolchildren with two or more doses of measles vaccine was 98% in the urban area, 92% in the rural area, and 85% in the border area (P < 0.05). In terms of opportunity, 90% of the children had received DTP1 + OPV1 before 3 months of age in the urban area, 89% had in the rural area, and 80% had in the border area (P < 0.05). The percentage of application of the complete basic schedule (DTP4 + OPV4 + MMR1) before 24 months of age was 93% in the urban area, 95% in the rural area, and 84% in the border area (P < 0.05). CONCLUSIONS: The border area had lower coverage of and opportunity for the basic schedule of vaccines, except for MMR. Follow-up campaigns for measles eradication have increased the coverage of the initial and booster doses in all three areas, but the increase has been greatest in the urban area. A greater effort should be made to identify children with an incomplete schedule of vaccinations, with priority going to areas that have a high proportion of immigrants. PMID- 15357938 TI - Antibiotic resistance in clinical isolates of Pseudomonas aeruginosa in Jamaica. AB - OBJECTIVE: To assess antibiotic resistance in clinical isolates of Pseudomonas aeruginosa in Jamaica, and to obtain baseline information on the presence of this important pathogen. METHODS: A total of 51 isolates of Pseudomonas aeruginosa, obtained from 162 clinical specimens from major hospitals and laboratories in seven parishes in Jamaica, were analyzed between May and August 2002. Isolates were tested against 18 different antibiotics by a disk diffusion method. RESULTS: Organisms were cultured from wound swabs (56%), high vaginal swabs (10.5%) and ear swabs (42.5%). Overall, the highest percentage rates of resistance were found for cefaclor (100% of all isolates), nalidixic acid (82.4%), kanamycin (76.5%), and trimethoprim/sulfamethoxazole (56.9%). Resistance rates were 25.5% or lower for tobramycin, gentamicin and polymyxin B, cefotaxime, ciprofloxacin and norfloxacin, piperacillin, carbapenems and amikacin. Forty-one isolates showed intermediate sensitivity to most of the antipseudomonal antibiotics, and the remaining 10 isolates were resistant to eight or more antibiotics. The multiresistant isolates, most of which were hospital isolates, were all resistant to tetracycline, nalidixic acid and trimethoprim/sulfamethoxazole, and highly (80%-90%) resistant to kanamycin, ciprofloxacin and norfloxacin. CONCLUSIONS: This study confirms that antibiotic resistance in this clinical pathogen is emerging in Jamaica, and suggests that due care must be taken in hospital settings to adequately diagnose pseudomonal infections and prescribe the antibiotic treatment most effective in preventing the increase in multidrug resistant organisms. PMID- 15357939 TI - [Clustering of modifiable cardiovascular risk factors in adults living in Salvador (BA), Brazil]. AB - OBJECTIVE: To estimate the frequency of modifiable cardiovascular risk factors, with and without inclusion of arterial hypertension, occurring simultaneously in a racially-mixed population. METHOD: A cross-sectional study was carried out with 1,298 adults aged > or = 20 years in the city of Salvador, Brazil, in 2000. Eight modifiable cardiovascular risk factors were assessed, in any combination: total cholesterol > or = 240 mg/dL; high density-lipoprotein cholesterol (HDL-c) < 40 mg/dL; triglycerides > or = 200 mg/dL; glycemia > or = 126 mg/dL + well controlled diabetes; body mass index > or = 25 kg/m2, waist > or = 102 cm for males and > or = 88 cm for females, smoking and alcoholism. The results were stratified according to the number of simultaneous risk factors (zero to five or more and two or more risk factors). The data were analyzed in terms of estimated proportions and 95% confidence intervals (95%CI), with and without the inclusion of arterial hypertension (VI Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure [JNC-VI], United States of America), ratio of proportions and chi-square for proportions as a measure of association. RESULTS: Among men (41.4% of participants), 7.5% (95%CI: 2.5 to 9.7) did not present risk factors; 68.8% (95%CI: 65.0 to 72.8) presented two or more risk factors, not including hypertension. After inclusion of hypertension, 73.4% (95%CI: 69.7 to 77.1) presented two or more risk factors. Among women, 11.6% did not present risk factors. The presence of two or more risk factors, not including hypertension, was observed in 67.7% (95%CI: 64.8 to 71.4). After inclusion of hypertension, 71.7% (95%CI: 68.5 to 74.9) of the women presented two or more risk factors. Significant differences were observed for the presence of two or more risk factors in men with not more than 4 years of schooling vs. 5 to less than 11 years of schooling (P < 0.05); in women with not more than 4 years of schooling vs. 5 to less than 11 years of schooling; in women with not more than 4 years of schooling vs. 11 or more years of schooling (P < 0.01); and in black vs. white women (P < 0.01). CONCLUSIONS: The high proportion of clustering cardiovascular risk factors in Salvador, with or without hypertension, especially in the population with little schooling and in black individuals, suggests the need for broad social strategies to reduce social inequality, promote health, and facilitate the treatment of cardiovascular risk factors. PMID- 15357940 TI - In Paraguay, both IUDs and contraceptive pills come with "strings attached". PMID- 15357941 TI - Intraepithelial lymphocytes in celiac disease: license to kill revealed. AB - Infiltration of lymphocytes in the small intestinal epithelium is a hallmark of active celiac disease. In this issue of Immunity, two papers uncover mechanisms controlling the cytolytic potential of these cells and provide evidence that lymphocyte killing of enterocytes is an important part of the celiac disease pathogenesis. PMID- 15357942 TI - Stop and go traffic to tune T cell responses. AB - Adaptive immune responses are initiated by interactions of T cells with antigen presenting cells, but the basic nature of these interactions during an immune response in vivo has been a matter of speculation. While some in vitro systems provide evidence for stable interactions, referred to as immunological synapses, compelling evidence supports T cell activation through serial transient interactions. Deep tissue intravital and organ culture microscopy studies suggest that both modes of interaction are employed, but new issues have emerged. This review will discuss in vitro results that framed the hypotheses that are currently being tested in vivo. I present a model in which TCR stop signals compete with chemokine-mediated go signals to adjust the duration of immunological synapse formation and tune the immune response between tolerance and full activation. PMID- 15357943 TI - Intravital microscopy: visualizing immunity in context. AB - Recent advances in photonics, particularly multi-photon microscopy (MPM) and new molecular and genetic tools are empowering immunologists to answer longstanding unresolved questions in living animals. Using intravital microscopy (IVM) investigators are dissecting the cellular and molecular underpinnings controlling immune cell motility and interactions in tissues. Recent IVM work showed that T cell responses to antigen in lymph nodes are different from those observed in vitro and appear dictated by factors uniquely relevant to intact organs. Other IVM models, particularly in the bone marrow, reveal how different anatomic contexts regulate leukocyte development, immunity, and inflammation. This article will discuss the current state of the field and outline how IVM can generate new discoveries and serve as a "reality check" for areas of research that were formerly the exclusive domain of in vitro experimentation. PMID- 15357944 TI - Illuminating the landscape of in vivo immunity: insights from dynamic in situ imaging of secondary lymphoid tissues. AB - A central feature of the immune system is the migratory behavior of its cellular components. Thus, fully understanding the generation and maintenance of immune responses must include consideration of how hematopoietic cells home to, interact within, and exit from secondary lymphoid organs as well as peripheral tissues. Recent advances in in situ imaging techniques now permit direct observation of these events in their physiologic settings with high spatiotemporal resolution. This review summarizes progress in this area of investigation from a lymphocentric perspective. We highlight controversies, point out key unanswered questions, and briefly outline what we believe are some of the near-term directions that in situ microscopic analysis of the immune system will take. PMID- 15357945 TI - Visualizing the first 50 hr of the primary immune response to a soluble antigen. AB - Recently, static and dynamic imaging methods have produced the first glimpses of the interactions between antigen-specific T cells and peptide-MHC-bearing antigen presenting cells in the lymph nodes. Using data from these experiments, we produced a numerically, spatially, and temporally scaled simulation of the first 50 hr of the primary T cell-dependent immune response. The simulation highlights how lymph node structure facilitates antigen presentation to rare, naive, antigen specific CD4+ T cells. PMID- 15357946 TI - Dynamic behavior of T cells and thymocytes in lymphoid organs as revealed by two photon microscopy. AB - The initial application of two-photon imaging to the study of lymphoid tissues has provided a dramatic glimpse into how cells of the immune system move and interact in their native environments. In this review we summarize what we have learned so far and point out areas for future investigation using this approach. PMID- 15357947 TI - Coordinated induction by IL15 of a TCR-independent NKG2D signaling pathway converts CTL into lymphokine-activated killer cells in celiac disease. AB - A major function of NKG2D linking innate and adaptive immunity is to upregulate antigen-specific CTL-mediated cytotoxicity in tissues expressing stress-induced NKG2D ligands, such as MIC, by coactivating TCR signaling. Here, we show that, under conditions of dysregulated IL15 expression in vivo in patients with celiac disease and in vitro in healthy individuals, multiple steps of the NKG2D/DAP10 signaling pathway leading to ERK and JNK activation are coordinately primed to activate direct cytolytic function independent of TCR specificity in effector CD8 T cells. These findings may not only explain previous reports of transformation of CTL into NK-like "lymphokine-activated killers" (LAK cells) under high doses of IL2 (a substitute for IL15) but may also have significant implications for understanding and treating immunopathological diseases. PMID- 15357948 TI - A direct role for NKG2D/MICA interaction in villous atrophy during celiac disease. AB - MICA molecules interact with the NKG2D-activating receptor on human NK and CD8 T cells. We investigated the participation of the MICA/NKG2D pathway in the destruction of intestinal epithelium by intraepithelial T lymphocytes (IEL) in Celiac disease and its premalignant complication, refractory sprue. We show that MICA is strongly expressed at epithelial cell surface in patients with active disease and is induced by gliadin or its p31-49 derived peptide upon in vitro challenge, an effect relayed by IL-15. This triggers direct activation and costimulation of IEL through engagement of NKG2D, leading to an innate-like cytotoxicity toward epithelial targets and enhanced TCR-dependent CD8 T cell mediated adaptive response. Villous atrophy in Celiac disease might thus be ascribed to an IEL-mediated damage to enterocytes involving NKG2D/MICA interaction after gliadin-induced expression of MICA on gut epithelium. This supports a key role for MIC/NKG2D in the activation of intraepithelial immunity in response to danger. PMID- 15357949 TI - B1b lymphocytes confer T cell-independent long-lasting immunity. AB - Many microbial pathogens employ antigenic variation as a strategy to evade the immune system, posing a challenge in vaccine development. To understand the requirements for immunity against such pathogens, we studied Borrelia hermsii, a relapsing fever bacterium. We found that mice deficient in T, follicular B, marginal zone B, or B1a lymphocytes resolved B. hersmii bacteremia and became resistant to reinfection. The resolution of bacteremia coincided with an expansion and persistence of B1b lymphocytes, and purified B1b lymphocytes from convalescent wild-type or TCR-betaxdelta-/- mice conferred immunity to Rag1-/- mice. The B1b lymphocytes in the reconstituted Rag1-/- mice provided long-lasting immunity by rapidly generating B. hermsii-specific IgM but not IgG upon bacterial challenge. Unmutated IgM is sufficient to eliminate B. hermsii, because AID-/- mice deficient in somatic hypermutation and class switch recombination efficiently resolved all bacteremic episodes. These data demonstrate that B1b lymphocytes can provide long-lasting T cell-independent IgM memory. PMID- 15357950 TI - Langerhans cells activate naive self-antigen-specific CD8 T cells in the steady state. AB - TCR transgenic mice that express a peptide antigen in keratinocytes develop a lethal CD8 T cell-dependent autoimmune disease. We employed an adoptive transfer system to understand this disease and show that transfer of low numbers of naive CD8 T cells into peptide transgenic mice caused chronic skin disease. The antigen presenting cell that initiated this response was the epidermal Langerhans cell. Naive CD8 T cells proliferated extensively, migrated to tissues, developed effector function, and were capable of making a recall response. These features are very different from the abortive activation of CD8 T cells that occurred in response to the same antigen presented by APC from other tissues. Furthermore, tolerance was dominant when the antigen was presented by both Langerhans cells and other APC. These data suggest that Langerhans cells do not have tolerogenic properties in the steady state. PMID- 15357951 TI - B7-1 and B7-2 selectively recruit CTLA-4 and CD28 to the immunological synapse. AB - The reported affinity differences between CD28 and CTLA-4 binding to B7-1 and B7 2 may serve to selectively regulate CD28 and CTLA-4 function by differentially recruiting and/or stabilizing these molecules at the immunological synapse. Here we show that ligand binding is important for the accumulation of both CD28 and CTLA-4 at the synapse. While CD28 is recruited to the synapse in the absence of B7-1 and B7-2 binding, it is not effectively stabilized there, as its localization can be disrupted by CTLA-4. In the case of CTLA-4, ligand binding is critical for its concentration at the synapse. We also demonstrate that the affinity and avidity differences in ligand binding translate into selective recruitment of CD28 or CTLA-4 to the immunological synapse--B7-1 is the major ligand mediating CTLA-4 localization, while B7-2 is the main ligand for CD28 concentration at the synapse. PMID- 15357952 TI - Compartmentalization of TNF receptor 1 signaling: internalized TNF receptosomes as death signaling vesicles. AB - The molecular regulation of the recruitment of initial signaling complexes at the TNF-R1 is poorly defined. We demonstrate here that within minutes internalized TNF-R1 (TNF receptosomes) recruits TRADD, FADD, and caspase-8 to establish the "death-inducing signaling complex" (DISC). In addition, we identified the TNF-R1 internalization domain (TRID) required for receptor endocytosis and provide evidence that TNF-R1 internalization, DISC formation, and apoptosis are inseparable events. Analyzing cell lines expressing an internalization-deficient receptor (TNF-R1 DeltaTRID) revealed that recruitment of RIP-1 and TRAF-2 to TNF R1 occurred at the level of the plasma membrane. In contrast, aggregation of TRADD, FADD, and caspase-8 to establish the TNF-R1-associated DISC is critically dependent on receptor endocytosis. Furthermore, fusion of TNF receptosomes with trans-Golgi vesicles results in activation of acid sphingomyelinase and cathepsin D. Thus, TNF receptosomes establish the different TNF signaling pathways by compartmentalization of plasma membrane-derived endocytic vesicles harboring the TNF-R1-associated DISC. PMID- 15357953 TI - Differential requirements for DOCK2 and phosphoinositide-3-kinase gamma during T and B lymphocyte homing. AB - Chemokines guide lymphocytes from blood to secondary lymphoid organs by triggering integrin-dependent firm adhesion under vascular flow and directed migration of T and B lymphocytes within lymphoid tissue. Here, we analyze the roles of DOCK2, a mammalian homolog of Caenorhabditis elegans CED-5 and Drosophila melanogaster Myoblast City, and phosphoinositide-3-kinase (PI3K) during lymphocyte recirculation. DOCK2 mediated efficient lymphocyte migration in a largely PI3K-independent manner, although a minor, PI3K-dependent pathway for migration was observed in wild-type and DOCK2-deficient lymphocytes. In T cells, this residual migration depended mainly on PI3Kgamma, whereas other PI3K isoforms were implicated in B cells. In vitro adhesion assays and intravital microscopy of lymphoid organ vasculature uncovered an unexpected defect in integrin activation in DOCK2-/- B cells, whereas lack of DOCK2 did not affect chemokine-triggered integrin activation in T cells. DOCK2 and PI3Kgamma thus play distinct roles during T and B cell integrin activation and migration. PMID- 15357954 TI - Regulation of BCR signal transduction in B-1 cells requires the expression of the Src family kinase Lck. AB - Although found predominantly in the peritoneal and pleural cavities, B-1 cells are also present in other peripheral tissues such as spleen and lung. While similar in surface phenotypes, such as CD5, all B-1 cells are not equivalent in their response to stimuli. Here, we report that the src family kinase Lck is required to confer the BCR hyporesponsiveness typical of CD5+ B-1 cells and appears involved in the maintenance of their unique function. Splenic B-1 cells express CD5 but not Lck and are not hyporesponsive; however, within the peritoneum, these B-1 cells are induced to express Lck and acquire a hyporesponsive phenotype. Peritoneal B-1 cells from lck-deficient mice, while CD5+, no longer exhibit attenuated BCR signaling. Interestingly, lck-null mice exhibited increased natural antibody levels characteristic of B-1 cells. Taken together, these results demonstrate a key role for Lck in modulating the signaling and cellular fate of B-1 B cells. PMID- 15357955 TI - Synthesis of a novel tetrahydroisoquinolino[2,1-c][1,4]benzodiazepine ring system with DNA recognition potential. AB - We report the first stereospecific synthesis and reactivity of a novel tetrahydroisoquinolino[2,1-c][1,4]benzodiazepine ring system with DNA recognition potential. PMID- 15357956 TI - Discovery of potent and orally bioavailable N,N'-diarylurea antagonists for the CXCR2 chemokine receptor. AB - A series of 3-substituted N,N'-diarylureas was prepared and the structure activity relationship relative to CXCR2 receptor affinity as well as their pharmacokinetic properties were examined. In vitro microsomal metabolism studies indicated that the lower clearance rates of the 3-sulfonamido-substituted compounds were most likely due to the suppression of glucuronidation. PMID- 15357957 TI - Synthesis, receptor binding and functional studies of mesoridazine stereoisomers. AB - The four stereoisomers of mesoridazine were synthesized and evaluated in D2, 5 HT1A, 5-HT2A, 5-HT2C, D1, and D3 receptor binding and functional assays. Two isomers demonstrated potent D2 receptor binding (Ki < 3 nM) and functional antagonism (IC50 < or = 10 nM) activities. These two isomers also showed moderate affinity for the 5-HT2A and D3 receptors. A third isomer was devoid of significant D2 receptor binding, but did have moderate affinity for the 5-HT2A and D3 receptors. The fourth isomer demonstrated poor affinity for all the receptors tested. Most significantly, the stereochemistry of the sulfoxide moiety played a dominant role in the observed structure-activity relationship (SAR). PMID- 15357958 TI - A library synthesis of 4-hydroxy-3-methyl-6-phenylbenzofuran-2-carboxylic acid ethyl ester derivatives as anti-tumor agents. AB - As a result of a hit-to-lead program using a technique of solution-phase parallel synthesis, a highly potent (2,4-dimethoxyphenyl)-[6-(3-fluorophenyl)-4-hydroxy-3 methylbenzofuran-2-yl]methanone (15b) was synthesized as an optimized derivative of 4-hydroxy-3-methyl-6-phenylbenzofuran-2-carboxylic acid ethyl ester (1), which was discovered as a screening hit from small-molecule libraries and exhibited selective cytotoxicity against a tumorigenic cell line. PMID- 15357959 TI - Synthesis of an unusual branched-chain sugar, 5-C-methyl-L-idopyranose for SAR studies of pyranmycins: implication for the future design of aminoglycoside antibiotics. AB - The syntheses of a challenging branched-chain sugar and several L-sugars have been accomplished. Their application in studies of the antibacterial activity of pyranmycins is reported, which could provide new strategies for the future design of aminoglycoside antibiotics. PMID- 15357960 TI - Identification of CJC-1131-albumin bioconjugate as a stable and bioactive GLP-1(7 36) analog. AB - A series of analogs of GLP-1(7-36) amide containing a Nepsilon-(2-[2-[2-(3 maleimidopropylamido)ethoxy]ethoxy]acetyl)lysine has been synthesized and the resulting derivatives were bioconjugated to Cys34 of human serum albumin (HSA). The GLP-1-HSA bioconjugates were analyzed in vitro to assess the stabilizing effect of bioconjugation in the presence of DPP-IV as well as GLP-1 receptor binding and activation. Compound 9 (CJC-1131) having the point of attachment to albumin at the C-terminal of GLP-1 and a D-alanine substitution at position 8 was identified as having the best combination of stability and bioactivity. PMID- 15357961 TI - Stereoselective synthesis and preliminary evaluation of new D-3 heteroarylcarbonylalanines as ligands of the NMDA receptor. AB - New N-heteroarylcarbonylalanines of the D-series were stereoselectively prepared from enoates derived from D-mannitol. These compounds were active in binding and functional assays of the NMDA sub-type of glutamate receptors. A pyridine derivative inhibited MK801 binding, protected neurons from excitotoxic damage and blocked NMDA-induced currents in neurons. A thiophene derivative positively modulated the NMDA receptor, possibly through the allosteric glycine site. PMID- 15357962 TI - Synthesis and structural-activity relationships of 3-hydroxyquinazoline-2,4-dione antibacterial agents. AB - A series of 3-hydroxyquinazoline-2,4-diones was synthesized and evaluated for antibacterial activity. This series represents a novel addition to the DNA gyrase inhibitor class of antibacterials. Appropriate substitutions onto the core template yielded compounds with excellent potency against E. coli gyrase and significant in vitro Gram-negative and Gram-positive antibacterial activity. PMID- 15357963 TI - Nonpeptide alpha V beta 3 antagonists. Part 9: Improved pharmacokinetic profile through the use of an aliphatic, des-amide backbone. AB - A series of alphaVbeta3 receptor antagonists lacking the amide bond of previously reported 'chain-shortened' compounds is described. Replacement of the lone amide bond with two methylene groups in this series yields more lipophilic compounds that have longer half-lives, lower clearance, and greater oral bioavailability when administered to dogs. PMID- 15357964 TI - Structure-activity relationships of piperazinebenzylamines as potent and selective agonists of the human melanocortin-4 receptor. AB - SAR studies on a series of piperazinebenzenes directed toward the human melanocortin-4 receptor resulted in potent MC4R agonists. Replacement of the triazole moiety of an initial lead 4 by a basic nitrogen baring a lipophilic side chain increased the binding affinities of these compounds. Analogs bearing an additional hetero-atom in the side-chain possessed good agonist potency. Thus, 11h had a Ki of 11 nM, and 13g exhibited an EC50 of 3.8 nM and a Ki of 6.4 nM. PMID- 15357965 TI - Structure-activity relationships of epolactaene derivatives: structural requirements for inhibition of Hsp60 chaperone activity. AB - Epolactaene is a microbial metabolite isolated from the fungal strain Penicillium sp. It arrests the cell cycle at the G0/G1 phase and induces the outgrowth of neurites in human neuroblastoma SH-SY5Y cells. In this communication, we report the structure-activity relationships (SARs) of new epolactaene derivatives, including those lacking the epoxylactam moiety and having various side chains. These derivatives were evaluated for their ability to inhibit the growth of human cancer cell lines. They were also analyzed for their ability to affect human heat shock protein 60 (Hsp60), which we have already identified as a protein that binds to epolactaene. We also identified the important structural framework of epolactaene/ETB (epolactaene tertiary butyl ester) for not only binding to Hsp60 but also inhibiting Hsp60 chaperone activity. PMID- 15357966 TI - Synthesis of 2',5'-oligoadenylate analogs containing an adenine acyclonucleoside and their ability to activate human RNase L. AB - This paper described synthesis of 2',5'-oligoadenylate (2-5A) analogs containing the purine acyclonucleoside, 9-[(2'S,3'R)-2',3',4'-trihydroxybutyl]adenine (2). The ability of the analogs to activate recombinant human RNase L was evaluated using 5'-32P-r(C11U2C7)-3' as a substrate. The EC50 value (the concentration of the 2-5A required to cleave half of the RNA) of the parent 2-5A tetramer 13 was 1.0 nM, whereas those of the analog 14 incorporating 2 at the second position from the 5'-end and the analog 15 incorporating 2 at the third position from the 5'-end were 9.0 and 1.7 nM, respectively. The analogs 14 and 15 were only 9- and 1.7-fold less potent than the parent 2-5A 13 itself, in RNase L activation ability. Furthermore, the oligodeoxynucleotide containing 2 was more resistant to nucleolytic hydrolysis by snake venom phosphodiesterase (a 3'-exonuclease) than the unmodified oligodeoxynucleotide. Thus, incorporation of an acyclonucleoside into 2-5A may be useful for developing an antiviral agent based on the 2-5A system. PMID- 15357967 TI - Synthesis and cellular uptake of a fluorescently labeled cyclic PNA-based compound. AB - A cyclic hexameric PNA-based compound labeled with fluorescein has been prepared following the liquid phase FPB strategy. Its cellular uptake, without and with electroporation, has been investigated by fluorescence microscopy. PMID- 15357968 TI - Design, synthesis and antimalarial activity of a glyoxylylhydrazone library. AB - Synthesis of a new family of quinolylhydrazone derivatives and evaluation of their activity against a chloroquine-resistant strain of Plasmodium falciparum are described. The best compound displayed an activity 6-fold higher than chloroquine. None of the active compounds were found to inhibit beta-hematin formation in vitro in the same range as chloroquine and five among them displayed lower calculated vacuolar accumulation ratios, suggesting the implication of a different mechanism of action. PMID- 15357969 TI - Discovery of inhibitors of human adipocyte fatty acid-binding protein, a potential type 2 diabetes target. AB - Low micromolar human A-FABP inhibitors were found by utilizing a fluorescence polarization assay, X-ray crystallography and modeling. The carbazole- and indole based inhibitors displayed approximately 10-fold preferences over human H-FABP and E-FABP, and are highly selective against I-FABP. This communication describes the SAR for drug-like synthetic inhibitors of human A-FABP. PMID- 15357970 TI - Substituted benzylamino-6-(trifluoromethyl)pyrimidin-4(1H)-ones: a novel class of selective human A-FABP inhibitors. AB - The synthesis and biological evaluation of novel human A-FABP inhibitors based on the 6-(trifluoromethyl)pyrimidine-4(1H)-one scaffold is described. Two series of compounds, bearing either an amino or carbon substituent in the 2-position of the pyrimidine ring were investigated. Modification of substituents and chain length optimization led to novel compounds with low micromolar activity and good selectivity for human A-FABP. PMID- 15357971 TI - Synthesis and structure-activity relationship of a novel sulfone series of TNF alpha converting enzyme inhibitors. AB - Replacement of the amide functionality in IM491 (N-hydroxy-(5S,6S)-1-methyl-6-[[4 (2-methyl-4-quinolinylmethoxy)anilinyl]carbonyl]5-piperidinecarboxamide) with a sulfonyl group led to a new series of alpha,beta-cyclic and beta,beta-cyclic gamma-sulfonyl hydroxamic acids, which were potent TNF-alpha converting enzyme (TACE) inhibitors. Among them, inhibitor 4b (N-hydroxy-(4S,5S)-1-methyl-5-[[4-(2 methyl-4-quinolinylmethoxy)phenyl]sulfonylmethyl]-4-pyrrolidinecarboxamide) exhibited IC50 values of < 1 nM and 180 nM in porcine TACE (pTACE) and cell assays, respectively, with excellent selectivity over MMP-1, -2, -9 and -13 and was orally bioavailable with an F value of 46% in mice. PMID- 15357972 TI - Synthesis and DP-IV inhibition of cyano-pyrazoline derivatives as potent anti diabetic agents. AB - A new series of cyano-pyrazoline derivatives with secondary amine at P-2 site was synthesized through achiral and chiral synthetic methods and evaluated for their ability to inhibit dipeptidyl peptidase IV (DP-IV). Compound 5i revealed good in vivo efficacy (ED50: 4.1 mg/kg; in vivo DP-IV inhibition). Also chiral derivative (11b) having (S)-configuration of compound 5i was found to be more potent. PMID- 15357973 TI - Ligand conformation has a definitive effect on 5-HT1A and serotonin reuptake affinity. AB - Conformationally constrained aryl cyclohexanes and cyclohexenes based on aryl cyclohexanols 1 were prepared. Locking the aryl ring in plane with the cyclohexane moiety provided potent SSRIs 3. Conversely, fixing the aryl ring perpendicular to the cyclohexane ring via a spiro lactone provided balanced 5 HT1A antagonists with mid-nanomolar range SSRI potency (compounds 2). PMID- 15357974 TI - Synthesis of chiral phosphoantigens and their activity in gamma delta T cell stimulation. AB - Gammadelta T cells expressing Vgamma2Vdelta2 T cell receptors are activated by a broad range of phosphorus-containing small molecules, termed phosphoantigens, and are of interest in the context of the chemotherapy of B cell malignancies. Here, we report the synthesis of four pairs of chiral phosphoantigens: the bromohydrins of isopentenyl diphosphate (Phosphostim), the epoxides of isopentenyl diphosphate (EIPP); and the corresponding bromohydrin and epoxide analogs of but-3-enyl diphosphate. The ability of each compound to stimulate human Vgamma2Vdelta2 T cells was determined by TNF-alpha release and cell proliferation. In these assays, the (R)-bromohydrin diphosphates were, on average, about twice as active as the (S)-bromohydrin diphosphates. In contrast, the (S)-form of EIPP was about twice as active as (R)-EIPP. The activities of the epoxy but-3-enyl diphosphates were both very low. These results suggest that chiral phosphoantigens, as opposed to racemic mixtures, may have utility in immunotherapy. PMID- 15357975 TI - 4-Aryl-3-(mercapto)quinolin-2-ones: novel maxi-K channel opening relaxants of corporal smooth muscle. AB - A novel series of 4-aryl-3-(mercapto)quinolin-2-one derivatives was prepared and evaluated as openers of the cloned maxi-K channel hSlo expressed in Xenopus laevis oocytes by utilizing electrophysiological methods. The effect of these maxi-K openers on corporal smooth muscle was studied in vitro using isolated rabbit corpus cavernosum. In vivo efficacy has been demonstrated with a selective maxi-K opening relaxant in a rat model of erectile function. PMID- 15357976 TI - Anti-HIV activity of some synthetic lignanolides and intermediates. AB - The evaluation of the anti-HIV-1 activity of synthetic lignanolides and their intermediates is reported. The antiviral activity was studied through luciferase based assays targeting the HIV-1 promoter activation induced by either, the HIV-1 Tat protein or the cellular transcription factor NF-kappaB, both known as crucial factors in HIV-1 replication. Among the compounds tested, three of them 2, 4 and 13 were further analysed for their anti-HIV-1 activity by recombinant virus assays, showing a suitable profile for development of novel anti-HIV-1 drugs. PMID- 15357977 TI - Cytotoxic effects and reversal of multidrug resistance by ibogan and related indole alkaloids. AB - A series of indole alkaloids of the ibogan-type was assessed for their cytotoxic effects as well as their potential in reversing MDR in vincristine-resistant KB cells. Of a total of 25 compounds tested, 3(S)-cyanocoronaridine, 3(S) cyanoisovoacangine, 3(S)-cyanovoacangine, and 10,11-demethoxychippiine were found to show appreciable cytotoxicity toward KB cells, while coronaridine, heyneanine, 19-epi-heyneanine, dippinine B, and dippinine C, were found to reverse MDR in vincristine-resistant KB cells. PMID- 15357978 TI - A suitable 1,2,4-oxadiazoles synthesis by microwave irradiation. AB - One pot microwave-assisted synthesis of substituted 1,2,4-oxadiazoles in solvent and under solvent free condition was performed exploring the importance of some coupling reagents. Good yields and short reaction times were the main aspects of the methods. PMID- 15357979 TI - Synthesis and antibacterial activity of C-6 carbamate ketolides, a novel series of orally active ketolide antibiotics. AB - A new series of antibacterial ketolides is reported, which features the use of a C-6 carbamate for tethering the arylalkyl sidechain to the macrolide core. The best members of this series display in vitro and in vivo activity comparable to telithromycin. Partial epimerization at C-2, unobserved in previously reported ketolides, was noted for this series. PMID- 15357980 TI - The purine transferase from Trypanosoma cruzi as a potential target for bisphosphonate-based chemotherapeutic compounds. AB - We identified and tested bisphosphonates as inhibitors of a protozoan molecular target. Computational modeling studies demonstrated that these compounds are mimics of the natural substrate of the enzyme. The most potent bisphosphonates in vitro are pamidronate and risedronate, which inhibit the purine transferase from Trypanosoma cruzi in the micromolar range. PMID- 15357981 TI - Isoindolinone ureas: a novel class of KDR kinase inhibitors. AB - A series of substituted isoindolinone ureas was prepared and evaluated for enzymatic and cellular inhibition of KDR kinase activity. Several of these analogs, such as 14c, are potent inhibitors of KDR both enzymatically (< 50 nM) and cellularly < or = 100 nM). A 3D KDR/CDK2/MAP kinase overlay model with several structurally related tyrosine kinase inhibitors was used to predict the binding interactions of the isoindolinone ureas with the KDR active site. PMID- 15357982 TI - Substituted 6-phenyl-pyridin-2-ylamines: selective and potent inhibitors of neuronal nitric oxide synthase. AB - The synthesis and nNOS and eNOS activity of 6-(4-(dimethylaminoalkyl)-/6-(4 (dimethylaminoalkoxy)-5-ethyl-2-methoxyphenyl)-pyridin-2-ylamines and 6-(4 (dimethylaminoalkyl)-/6-(4-(dimethylaminoalkoxy)-2,5-dimethoxyphenyl)-pyridin-2 ylamines 1-8 are described. These compounds are potent inhibitors of the human nNOS isoform. PMID- 15357983 TI - Nonpeptide alpha V beta 3 antagonists. Part 10: In vitro and in vivo evaluation of a potent 7-methyl substituted tetrahydro-[1,8]naphthyridine derivative. AB - Subtle modifications were incorporated into the structure of clinical candidate 1. These changes were designed to maintain potency and selectivity while inducing changes in physical properties leading to improved pharmacokinetics in three species. This approach led to the identification of 4 as a potent, selective alphaVbeta3 receptor antagonist that was selected for clinical development based on an improved PK profile and efficacy demonstrated in an in vivo model of bone turnover. PMID- 15357984 TI - Syntheses and anti-MRSA activities of the C3 analogs of mansonone F, a potent anti-bacterial sesquiterpenoid: insights into its structural requirements for anti-MRSA activity. AB - Syntheses and excellent anti-MRSA activities of the mansonone F analogs are reported. In addition, the minimal structural requirements for its anti-MRSA activities as well as its structure-activity relationship including the C3 substituents effects on anti-MRSA activity are also described. In particular, this study revealed that both ortho-quinone and tricyclic systems of mansonone F are essential for anti-MRSA activities. PMID- 15357985 TI - AL-12182, a novel 11-oxa prostaglandin analog with topical ocular hypotensive activity in the monkey. AB - A series of 11-oxa prostaglandin analogs was evaluated for FP receptor binding and activation. Several compounds having aryloxy-terminated lower chains were found to be potent agonists. Topical ocular dosing of AL-12182, the isopropyl ester prodrug of the potent agonist 13, lowered intraocular pressure in the monkey by 40% accompanied by minimal conjunctival hyperemia in the rabbit. AL 12182 was synthesized on multigram scale starting with D-sorbitol. PMID- 15357986 TI - Antibody light chain-catalyzed hydrolysis of a hepatitis C virus peptide. AB - A panel of human monoclonal and recombinant antibody light chains was screened for cleavage of the synthetic peptide corresponding to a neutralizing epitope of hepatitis C virus (residues 192-205 of envelope glycoprotein E1). One of the 39 light chains studied hydrolyzed the Val197-Ser198 bond of the peptide with Km and kcat values of 223 +/- 7 microM and 0.087 +/- 0.001 min(-1). PMID- 15357987 TI - Synthesis and KCNQ2 opener activity of N-(1-benzo[1,3]dioxol-5-yl-ethyl, N-[1 (2,3-dihydro-benzofuran-5-yl)-ethyl, and N-[1-(2,3-dihydro-1H-indol-5-yl)-ethyl acrylamides. AB - Bioisosteric replacement studies led to the identification of N-(1 benzo[1,3]dioxol-5-yl-ethyl)-3-(2-chloro-phenyl)-acrylamide ((S)-3) as a highly potent KCNQ2 opener, and 3-(2,6-difluoro-phenyl)-N-[1-(2,3-dihydro-benzofuran-5 yl)-ethyl]-acrylamide ((S)-4), and N-[1-(2,3-dihydro-1H-indol-5-yl)-ethyl]-3-(2 fluoro-phenyl)-acrylamide ((S)-5) as highly efficacious KCNQ2 openers. In contrast, their respective R enantiomers showed significantly less or no appreciable KCNQ2 opener activity even at the highest concentration tested (10 microM). Because of its high potency and moderate efficacy as well as its convenient synthesis, (+/-)-3 was selected as a reference compound for analyzing efficacies of KCNQ openers in electrophysiology studies. Compounds (S)-4 and (S) 5 demonstrated significant activity in reducing neuronal hyperexcitability in rat hippocampal slices. The synthesis and the KCNQ2 opener activity of these acrylamides are described. PMID- 15357988 TI - Evaluation of pyrrolidin-2-imines and 1,3-thiazolidin-2-imines as inhibitors of nitric oxide synthase. AB - Syntheses and evaluation of pyrrolidin-2-imines and 1,3-thiazolidin-2-imines as inhibitors of nitric oxide synthase (NOS) are discussed. An extensive SAR was established for pyrrolidin-2-imines class of compounds. The amidines came out as the most potent inhibitors in addition to displaying selectivity. PMID- 15357989 TI - Novel bisbenzamidines as potential drug candidates for the treatment of Pneumocystis carinii pneumonia. AB - A series of pentamidine congeners has been synthesized and screened for their in vitro activity against Pneumocystis carinii. Among the tested compounds, bisbenzamidines linked by a flexible pentanediamide or hexanediamide chain (7 and 9) emerged as exceptionally potent agents that were more effective and less toxic than pentamidine in the assays described in this study. PMID- 15357990 TI - Synthesis and neurotrophic activity of nonimmunosuppressant cyclosporin A derivatives. AB - In order to exploit cyclophilin as a potential target for neurological drug design, we demonstrate in this presentation that several nonimmunosuppressant analogues of cyclosporin A, modified at the various positions in the 'effector' domain, are equipotent nerve growth agents compared to cyclosporin A. Our results suggest that neurotrophic activity of cyclosporin A and its derivatives resides in the binding domain, and binding to cyclophilin and/or inhibiting rotamase activity may be a necessity for neurotrophic effects of cyclophilin ligands. PMID- 15357991 TI - Design, synthesis, and biochemical evaluation of novel alpha V beta 3 integrin ligands. AB - Studies on integrin alphaVbeta3 have implicated this receptor in a number of pathologies. In this article we describe some of our initial efforts to design small molecules alphaVbeta3 ligands incorporating an indole core template and an oxyguanidine as basic ending. Synthesis, biochemical activity and pharmacological properties are analyzed. PMID- 15357992 TI - Discovery of new chemical leads for prostaglandin D2 receptor antagonists. AB - A series of Indomethacin analogs were synthesized and biologically evaluated. Among the compounds tested, N-(p-butoxy)benzoyl-2-methylindole-4-acetic acid 2 was discovered as a new chemical lead for a prostaglandin D2 (PGD2) receptor antagonist. Structure-activity relationship data are also presented. PMID- 15357993 TI - Carbonic anhydrase inhibitors. Inhibition of the beta-class enzyme from the methanoarchaeon Methanobacterium thermoautotrophicum (Cab) with anions. AB - The first inhibition study of the beta-class carbonic anhydrase (CA, EC 4.2.1.1) from the methanoarchaeon Methanobacterium thermoautotrophicum (Cab) with anions is reported here. Inhibition data of the alpha-class human isozymes hCA I and hCA II (cytosolic) as well as the membrane-bound isozyme hCA IV and the gamma-class enzyme from another archaeon, Methanosarcina thermophila (Cam) with a large number of anionic species such as halides, pseudohalides, bicarbonate, carbonate, nitrate, nitrite, hydrosulfide, bisulfite, sulfate, etc., are also provided for comparison. The best Cab anion inhibitors were thiocyanate and hydrogen sulfide, with inhibition constants in the range of 0.52-0.70 mM, whereas cyanate, iodide, carbonate, and nitrate were weaker inhibitors (Ki's in the range of 7.8-13.2 mM). Fluoride, chloride, and sulfate do not inhibit this enzyme appreciably, whereas the CA substrate bicarbonate, or other anions, such as bromide, nitrite, bisulfite, or sulfamate behave as weak inhibitors (Ki in the range of 40-45 mM). It is interesting to note that the metal poison, coordinating anions cyanide and azide are also rather weak Cab inhibitors (Ki in the range of 27-55 mM), whereas sulfamide is a very weak Cab inhibitor (Ki of 103 mM), although it strongly inhibits Cam (Ki of 70 microM). Surprisingly, phenylboronic and phenylarsonic acids, which have been investigated for the inhibition of all these CAs for the first time, showed very weak activity against the alpha-CA isozymes, but were effective Cab and Cam inhibitors. The best Cab inhibitors were just these two compounds (Ki's of 0.20-0.33 mM), whereas the best Cam inhibitor was sulfamic acid (Ki of 96 nM). These major differences of behavior between the diverse CAs investigated here toward anion inhibitors can be difficultly explained considering the convergent evolution of so diverse enzymes for the binding and turnover of small molecules such as carbon dioxide and anions. PMID- 15357994 TI - Possible differences in modes of agonist and antagonist binding at human 5-HT6 receptors. AB - A graphics model of the human 5-HT6 receptor was constructed and automated docking studies were performed. The model suggests that 5-HT6 antagonist arylsulfonyltryptamines might bind differently than that of the agonist serotonin. Furthermore, the model explains many of the empirical results from our previous structure-affinity studies. PMID- 15357995 TI - SAR and pharmacokinetic studies on phenethylamide inhibitors of the hepatitis C virus NS3/NS4A serine protease. AB - SAR on the phenethylamide 1 (Ki 1.2 microM) in the P2- and the P'-position led to potent inhibitors, one of which showed good exposure and low clearance when administered intramuscularly to rat. PMID- 15357996 TI - Chemoenzymatic synthesis of (S) and (R)-propranolol and sotalol employing one-pot lipase resolution protocol. AB - Synthesis of both enantiomers of biologically active propranolol and sotalol has been achieved in high optical purity by one-pot reduction of 3 and 7 followed by in situ lipase resolution of the respective chlorohydrins. Pseudomonas cepacia lipase immobilized on ceramic particles (PS-C) provided the chlorohydrin and acetate, which on nucleophilic substitution with isopropyl amine afforded the target amino alcohols in high enantioselectivity under mild reaction conditions. PMID- 15357997 TI - Non-peptide oxytocin agonists. AB - A library of compounds targeted to the vasopressin/oxytocin family of receptors was screened for activity at a cloned human oxytocin receptor using a reporter gene assay. Potency and selectivity were optimised to afford compound 39, EC50 = 33 nM. This series of compounds represents the first disclosed, non-peptide, low molecular weight agonists of the hormone oxytocin (OT). PMID- 15357999 TI - Trends and innovations in industrial biocatalysis for the production of fine chemicals. AB - Biocatalysis has become an established technology for the industrial manufacture of fine chemicals. In recent years, a multitude of chemical companies have embraced biocatalysis for the manufacture of desired stereoisomers, and new or improved methods for the synthesis of enantiomerically pure alpha- and beta-amino acids, amines, amides, peptides, nitriles, alcohols, organic acids and epoxides have emerged. Furthermore, the selectivity and mild operational conditions of biocatalysts are increasingly applied in industry to modify complex target molecules. These recent innovations in the manufacture of industrial fine chemicals using biocatalysis are discussed from an industrial perspective. PMID- 15358000 TI - Enantioselective biocatalysis optimized by directed evolution. AB - Directed evolution methods are now widely used for the optimization of diverse enzyme properties, which include biotechnologically relevant characteristics like stability, regioselectivity and, in particular, enantioselectivity. In principle, three different approaches are followed to optimize enantioselective reactions: the development of whole-cell biocatalysts through the creation of designer organisms; the optimization of enzymes with existing enantioselectivity for process conditions; and the evolution of novel enantioselective biocatalysts starting from non-selective wild-type enzymes. PMID- 15358001 TI - Enzyme assays for high-throughput screening. AB - Assaying enzyme-catalyzed transformations in high-throughput is crucial to enzyme discovery, enzyme engineering and the drug discovery process. In enzyme assays, catalytic activity is detected using labelled substrates or indirect sensor systems that produce a detectable spectroscopic signal upon reaction. Recent advances in the development of high-throughput enzyme assays have identified new labels and chromophores to detect a wide range of enzymes activities. Enzyme activity profiling and fingerprinting have also been used as tools for identification and classification, while microarray formats have been devised to increase throughput. PMID- 15358002 TI - Biotechnological applications of hydrogenases. AB - Hydrogenases have found use in a variety of biotechnological applications, including biohydrogen production, wastewater treatment, the prevention of microbial-induced corrosion and the generation and regeneration of NADP cofactors. In the future, advances in genome mining and screening techniques are likely to identify new hydrogenases for novel applications. PMID- 15358003 TI - Improved beta-lactam acylases and their use as industrial biocatalysts. AB - Whereas the beta-lactam acylases are traditionally used for the hydrolytic processing of penicillin G and cephalosporin C, new and mutated acylases can be used for the hydrolysis of alternative fermentation products as well as for the synthesis of semisynthetic beta-lactam antibiotics. Three-dimensional structural analyses and site-directed mutagenesis studies have increased the understanding of the catalytic mechanism of these enzymes. The yield of hydrolysis and synthesis has been greatly improved by process design, including immobilization of the enzyme and the use of alternative reaction media. Significant advances have also been made in the resolution of racemic mixtures by means of stereoselective acylation/hydrolysis using beta-lactam acylases. PMID- 15358004 TI - Three-dimensional structures of enzymes useful for beta-lactam antibiotic production. AB - Significant advances have been made in the structure-based engineering of enzymes useful for beta-lactam antibiotic production. Structure-based engineering of penicillin G acylase and cephalosporin acylase has resulted in improved enzymes for use in enzymatic production processes. The structures of many other enzymes that could be used in the production of beta-lactam antibiotics, such as enzymes from the beta-lactam biosynthetic pathway and beta-lactam antibiotic-converting enzymes, have been determined. The interest in these structures suggests that the future may see an even more extensive use of rationally engineered biocatalysts in antibiotic production than today. PMID- 15358007 TI - The chick embryo--past, present and future as a model system in developmental biology. PMID- 15358008 TI - From the hemangioblast to self-tolerance: a series of innovations gained from studies on the avian embryo. AB - During the last decades of the 20th century, studies on the vertebrate hematopoietic and immune systems have largely been performed, on mammalian models. The mouse has been the preferred material for several cogent reasons: (i) numerous well defined genetic strains are available; (ii) this species has been and still is instrumental in the study of gene activity through transgenesis; and (iii) in vitro culture techniques and in vivo assays for blood cells together with a wide array of antibodies and nucleic acid probes have been developed to investigate the cellular interactions occurring during hematopoiesis and immune reactivity. However, important and fundamental notions have emerged from using another higher vertebrate model, the avian embryo. The distinction among small lymphocytes of two populations, the T and B lymphocytes, endowed with different roles in adaptive immunity and dependant on different environments for their specification, has relied on experiments carried out in birds. The avian model has been critical for the analysis of the origin and traffic of hematopoietic precursor cells. It allowed the demonstration that both hematopoietic and angioblastic lineages arise from a common precursor, a cell whose existence had been proposed but never undoubtedly proven, the hemangioblast. Finally a form of thymus-dependant 'dominant' tolerance was demonstrated on the basis of experiments in the avian embryo, which initiated a large current of studies on 'regulatory T-cells'. Work in this model during the last decades has relied strongly on the construction of chimeras between quail and chick embryos that allowed a refined analysis of cell behaviour during embryogenesis. The novel perception of developmental neuropoiesis and immunopoiesis that followed proved to be largely applicable to lower and higher vertebrates, notably mammals. PMID- 15358010 TI - Therapeutic update: non-selective beta- and alpha-adrenergic blockade in patients with coexistent chronic obstructive pulmonary disease and chronic heart failure. AB - Patients with chronic heart failure (CHF) have a resting restrictive ventilatory defect. Any type of exercise requires patients with CHF to markedly increase their minute ventilation. Patients with chronic obstructive pulmonary disease (COPD) have airflow obstruction that leads to dynamic lung hyperinflation and reduced ventilatory response to exercise. Because exercise is associated with abnormally high minute ventilation in patients with CHF and with a limited minute ventilation increase in patients with COPD, functional capacity is severely impaired in patients with coexistent CHF and COPD. Optimal treatment of both conditions is a prerequisite to maximally improve functional capacity in patients with CHF and COPD. Unfortunately, beta-adrenergic blockade, the current cornerstone of CHF therapy, is frequently omitted in patients with CHF and COPD for fear of inducing bronchoconstriction. Furthermore, when prescribed, beta adrenergic blockade is often attempted with a moderate dose of metoprolol tartrate, a beta-1-blocker that results in lesser clinical benefits than combined non-selective beta-blockade with carvedilol at the maximally recommended dose. Recent experience indicates that combined non-selective beta- and alpha-blockade with carvedilol is well tolerated in patients with COPD who do not have reversible airway obstruction. Alpha-adrenergic blockade may promote mild bronchodilation that offsets non-selective beta blockade-induced bronchoconstriction in patients with obstructive airway disease. PMID- 15358011 TI - Disparities in cardiac care: rising to the challenge of Healthy People 2010. AB - Eliminating health disparities is one of two overarching goals of Healthy People 2010. Although the causes of health disparities are complex, they appear to be related, in part, to disparities in the quality of medical care. Two recent reviews of peer-reviewed research investigated the evidence on racial/ethnic differences in medical care. An Institute of Medicine summary of the literature concluded that in most studies, racial and ethnic disparities in health care remained even after adjustment for potentially confounding factors. A review focused specifically on cardiac care, conducted jointly by the Kaiser Family Foundation and the American College of Cardiology Foundation, reached a similar conclusion after examining the most rigorous studies investigating racial/ethnic differences in angiography, angioplasty, coronary artery bypass graft (CABG) surgery, and thrombolytic therapy. For example, African Americans were statistically less likely than whites to undergo CABG surgery in 21 of the 23 most rigorous studies that calculated odds ratios to compare CABG use. Although there is a convincing body of evidence that race continues to matter in the health system, a nationally representative survey of physicians revealed that the majority of physicians do not view a patient's race/ethnicity as a factor in obtaining care, but do believe insurance coverage matters. Increasing physicians' awareness of the evidence for the role that race/ethnicity plays in health care is important because they are in a good position to directly and indirectly affect changes in clinical practice or patient behavior that could reduce disparities in care. PMID- 15358012 TI - Cardiovascular therapies and risk for development of diabetes. AB - The prevalence of diabetes is increasing, and patients with diabetes are at increased risk of adverse cardiovascular outcomes. Recently, the results from 11 large randomized clinical trials have suggested a difference in the emergence of new diabetes according to cardiovascular medication use. Treatment with angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, and calcium antagonists yielded a lower incidence of diabetes development than beta blockers and diuretics. Physicians should consider this possible diabetes consequence when prescribing long-term beta-blockers and diuretics, particularly in patients at high risk of developing diabetes. PMID- 15358013 TI - Multicenter USA Amplatzer patent ductus arteriosus occlusion device trial: initial and one-year results. AB - OBJECTIVES: We sought to review and report initial and one-year efficacy and safety results of the multicenter USA Amplatzer ductal occluder (ADO) device trial. BACKGROUND: Transcatheter closure of a moderate to large patent ductus arteriosus (PDA) using conventional techniques is challenging. The ADO can close a PDA up to 12 mm in diameter. METHODS: From September 1999 to June 2002, 484 patients were enrolled in 25 U.S. centers. Forty-five (9%) of 484 patients did not have ADO implantation, because the PDA was too small or because of elevated pulmonary resistance. The median age of the patients at catheterization was 1.8 years (range 0.2 to 70.7 years), and weight was 11 kg (range 4.5 to 164.5 kg). RESULTS: The median PDA minimal diameter was 2.6 mm (range 0.9 to 11.2 mm); 76 (17%) of 439 were larger than 4.0 mm. Median pulmonary artery mean pressure was 20 mm Hg (range 7 to 80 mm Hg). The ADO was implanted successfully in 435 (99%) of 439 patients, with a median fluoroscopy time of 7.1 min (range 2.9 to 138.4 min). Angiographic demonstration of occlusion was seen in 329 (76%) of 435. This increased to 384 (89%) of 433 on post-catheterization day 1, with occlusion documented in 359 (99.7%) of 360 at one year. At the last evaluation in all patients at any time, PDA closure was documented in 428 (98%) of 435 patients. There have been two cases of partial left pulmonary artery occlusion after ADO implantation and no cases of significant aortic obstruction. CONCLUSIONS: Moderate to large PDAs can be effectively and safely closed using the ADO device, with excellent initial and one-year results. This device should obviate the need for multiple coils or surgical intervention for these defects. PMID- 15358014 TI - Intracoronary brachytherapy after stenting de novo lesions in diabetic patients: results of a randomized intravascular ultrasound study. AB - OBJECTIVES: We studied the efficacy of intracoronary brachytherapy (ICB) after successful coronary stenting in diabetic patients with de novo lesions. BACKGROUND: Intracoronary brachytherapy has proven effective in preventing recurrences in patients with in-stent restenosis. However, the role of ICB for the treatment of de novo coronary stenoses remains controversial. METHODS: Ninety two patients were randomized to either ICB or no radiation after stenting. Primary end points were in-stent mean neointimal area (primary end point of efficacy) and minimal luminal area of the entire vessel segment (primary end point of effectiveness), as assessed by intravascular ultrasound at six-month follow-up. Quantitative coronary angiography analysis was performed at the target, injured, irradiated, and entire vessel segments. RESULTS: At follow-up, the in-stent mean neointimal area was 52% smaller in the ICB group (p < 0.0001). However, there was no difference in the minimal luminal area of the vessel segment (4.5 +/- 2.4 mm2 vs. 4.4 +/- 2.1 mm2). Restenosis rates increased progressively by the analyzed segment in the ICB group: target (7.1% vs. 20.9%, p = 0.07), injured (9.5% vs. 20.9%, p = NS), irradiated (14.3% vs. 20.9%, p = NS), and vessel segment (23.8% vs. 25.6%, p = NS). At one year, 1 cardiac death, 6 myocardial infarctions (MIs) (3 due to late stent thrombosis), and 10 target vessel revascularizations (TVRs) (6 due to the edge effect) occurred in the ICB group, whereas in the nonradiation group, there were 11 TVRs and no deaths or MIs. CONCLUSIONS: Intracoronary brachytherapy significantly inhibited in-stent neointimal hyperplasia after stenting in diabetic patients. However, clinically this was counteracted by the occurrence of the edge effect and late stent thrombosis. PMID- 15358015 TI - Direct stenting versus direct stenting followed by centered beta-radiation with intravascular ultrasound-guided dosimetry and long-term anti-platelet treatment: results of a randomized trial: Beta-Radiation Investigation with Direct Stenting and Galileo in Europe (BRIDGE). AB - OBJECTIVES: We sought to assess the efficacy of vascular brachytherapy (VBT) combined with stenting for the primary prevention of restenosis. BACKGROUND: Intravascular brachytherapy after stent implantation for de novo lesions has been abandoned for the present. We revisited this procedure by optimizing all procedural steps-the use of glycoprotein IIb/IIa blockers, direct stenting, adequate radiation coverage, avoidance of edge damage, source centering, intravascular ultrasound-guided dosimetry, and continuation of a dual anti platelet regimen for one year. METHODS: The Beta-Radiation Investigation with Direct stenting and Galileo in Europe (BRIDGE) study is a multicenter, randomized controlled trial evaluating the long-term efficacy of VBT with P-32 (20 Gy at 1 mm in the coronary wall) after direct stenting. The primary end point was angiographic intra-stent late loss; secondary end points were six months binary restenosis and neo-intimal hyperplasia. Patients (n = 112) with de novo lesions (2.5 to 4.0 mm in diameter up to 15 mm long) were randomized to either VBT or no VBT. RESULTS: At six months, intra-stent loss was 0.43 and 0.84 mm (p < 0.001) in the irradiated and control groups, respectively. Intra-stent neo-intimal volume was reduced from 36 mm3 to 10 mm3. However, in the irradiated group there were six late occlusions as well as eight restenoses outside the stented and peri stented area at the fall-off dose edges of the irradiated area. Accordingly, the target vessel revascularization and major adverse cardiac and cerebrovascular events rates at one year in the VBT group (20.4% and 25.9%, respectively) were higher than in the control group (12.1% and 17.2%, respectively). CONCLUSIONS: Despite the optimization of pre-, peri-, and post-procedural factors and despite the relative efficacy of the brachytherapy for the prevention of the intra-stent neo-intimal hyperplasia, the clinical outcome of the irradiated group was less favorable than that of the control group. PMID- 15358016 TI - Vascular brachytherapy and coronary stenting for de novo lesions: love on the rocks. PMID- 15358017 TI - Anemia is an independent predictor of mortality after percutaneous coronary intervention. AB - OBJECTIVES: The aim of the present study was to assess whether anemia is a marker of increased risk during interventional procedure and poor midterm survival after percutaneous coronary intervention (PCI). BACKGROUND: Anemia is associated with increased risk of mortality in patients with heart failure and myocardial infarction (MI). METHODS: We examined the outcomes of 6,116 consecutive PCI patients based on the hemoglobin (Hb) value before the interventional procedure. Patients were divided into three groups based on the baseline Hb level (g/l): Hb <10 = severe anemia; Hb 10 to 12 = mild anemia; Hb >12 = no anemia. RESULTS: The presence of anemia is associated with higher 30-day major adverse cardiac events, post-PCI peak troponin and creatine kinase-MB fraction, and a longer length of stay. After controlling for multiple covariates, significant difference in one year survival was noted in the anemic groups compared with no anemia group (adjusted hazard ratio for Hb 10 to 12: 1.5 [95% confidence interval 1.3 to 1.8]; for Hb <10: 1.8 [95% confidence interval 1.3 to 2.3]; p = 0.004.) This adverse effect of anemia on survival was noted in all three presenting clinical syndromes (stable angina, unstable angina, and MI). CONCLUSIONS: Anemia is an independent predictor of mortality after PCI and is associated with higher short-term adverse procedural events. PMID- 15358018 TI - Impact of anemia in patients with acute myocardial infarction undergoing primary percutaneous coronary intervention: analysis from the Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications (CADILLAC) Trial. AB - OBJECTIVES: We sought to investigate the impact of anemia in patients with acute myocardial infarction (AMI) undergoing primary percutaneous coronary intervention (PCI). BACKGROUND: The prognostic importance of anemia on primary PCI outcomes is unknown. METHODS: In the Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications (CADILLAC) trial, 2,082 patients of any age with AMI within 12 h onset undergoing primary PCI were randomized to balloon angioplasty versus stenting, each +/- abciximab. Outcomes were stratified by the presence of anemia at baseline, as defined by World Health Organization criteria (hematocrit <39% for men and <36% for women). RESULTS: Anemia was present in 260 (12.8%) of 2,027 randomized patients with baseline laboratory values. Patients with versus without baseline anemia more frequently developed in-hospital hemorrhagic complications (6.2% vs. 2.4%, p = 0.002), had higher rates of blood product transfusions (13.1% vs. 3.1%, p < 0.0001), and had a prolonged (median 4.1 vs. 3.5 days, p < 0.0001) and more expensive (median costs $12,434 vs. $11,603, p = 0.002) index hospitalization. Patients with versus without anemia had strikingly higher mortality during hospitalization (4.6% vs. 1.1%, p = 0.0003), at 30 days (5.8% vs. 1.5%, p < 0.0001), and at 1 year (9.4% vs. 3.5%, p < 0.0001). The rates of disabling stroke at 30 days (0.8% vs. 0.1%, p = 0.005) and at 1 year (2.1% vs. 0.4%, p = 0.0007) were also significantly higher in patients with anemia. By multivariate analysis, anemia was an independent predictor of in-hospital mortality (hazard ratio, 3.26; p = 0.048) and one-year mortality (hazard ratio, 2.38; p = 0.016). CONCLUSIONS: Anemia at baseline in patients with AMI undergoing primary PCI is common, and is strongly associated with adverse outcomes and increased mortality. PMID- 15358019 TI - The pathologic basis of Q-wave and non-Q-wave myocardial infarction: a cardiovascular magnetic resonance study. AB - OBJECTIVES: The purpose of this study was to determine the pathologic basis of Q wave (QW) and non-Q-wave (NQW) myocardial infarction (MI). BACKGROUND: The QW/NQW distinction remains in wide clinical use but the meaning of the difference remains controversial. We hypothesized that measurement of total MI size and transmural extent by late gadolinium enhancement cardiovascular magnetic resonance (CMR) would identify the pathologic basis of QWs. METHODS: A total of 100 consecutive patients with documented previous MI had electrocardiogram and CMR on the same day. Patients with acute MI within seven days were excluded. Left ventricular function and the size and transmural extent of MI were quantified in the three major arterial territories and correlated with the presence of QW. RESULTS: Subendocardial MI showed QW in 28%. Transmural MI showed NQW in 29%. Of all MIs, 48% were at some point transmural, and 99% of these were at some point non-transmural. As MI size and number of transmural segments increased, the probability of QW increased (anterior: total size chi-square = 53, p < 0.0001, transmural extent chi-square = 36, p < 0.0001; inferior: total size chi-square = 16, p = 0.001, transmural extent chi-square = 10, p = 0.001). These findings did not hold for lateral MI. In a multivariate model, the transmural extent of MI was not an independent predictor of QW when total size of MI was removed. The QW/NQW classification was a good test for size of MI (area under receiver operating characteristic curve: anterior 0.90, inferior 0.77). CONCLUSIONS: The QW/NQW distinction is useful, but it is determined by the total size rather than transmural extent of underlying MI. PMID- 15358020 TI - Q-wave and non-Q-wave myocardial infarctions through the eyes of cardiac magnetic resonance imaging. PMID- 15358021 TI - Association of elevated B-type natriuretic peptide levels with angiographic findings among patients with unstable angina and non-ST-segment elevation myocardial infarction. AB - OBJECTIVES: We hypothesized that elevated B-type natriuretic peptide (BNP) levels would be associated with a greater severity of angiographic disease and a greater extent of myocardium at risk. BACKGROUND: Elevations of BNP have been associated with increased risk of adverse outcomes in patients with unstable angina and non ST-segment elevation myocardial infarction (UA/NSTEMI). METHODS: Of the 2,220 patients with UA/NSTEMI enrolled in the Treat Angina with Aggrastat and Determine Cost of Therapy with an Invasive or Conservative Strategy-Thrombolysis In Myocardial Infarction-18 (TACTICS-TIMI-18) trial, 276 randomized to the invasive arm had both baseline BNP levels and angiographic core laboratory data. Patients were categorized according to their baseline BNP levels as < or =80 or >80 pg/ml. RESULTS: A total of 233 patients (84%) had BNP levels >80 pg/ml, and 43 (16%) had admission BNP levels >80 pg/ml. Patients with BNP >80 pg/ml had tighter culprit vessel stenosis on quantitative coronary angiography (median stenosis 76% vs. 67%, p = 0.004) and a higher (slower) corrected TIMI frame count (median CTFC 43 vs. 30, p = 0.018) in the culprit vessel. The median BNP level was higher in patients with a left anterior descending coronary artery (LAD) versus non-LAD culprit lesion location (median BNP level 40 vs. 24 pg/ml, p = 0.005), and the culprit artery was more often the LAD in patients with BNP >80 pg/ml compared with < or =80 pg/ml (44% vs. 30%, p = 0.06). CONCLUSIONS: Among patients with UA/NSTEMI, elevated BNP levels are associated with tighter culprit stenosis, higher CTFC, and LAD involvement. These findings suggest that elevated BNP may be associated with a greater severity and extent of myocardial ischemic territory during the index event and may partly explain the association between elevated BNP and adverse outcomes. PMID- 15358022 TI - Cardiac troponin after major vascular surgery: the role of perioperative ischemia, preoperative thallium scanning, and coronary revascularization. AB - OBJECTIVES: We sought to determine the role of preoperative predictors, particularly ischemia, on preoperative thallium scanning (PTS) and coronary revascularization on low-level and conventional troponin elevations after major vascular surgery. BACKGROUND: Postoperative cardiac troponin (cTn) elevations have recently been shown to predict both short- and long-term mortality after vascular surgery. METHODS: The perioperative data, including PTS and subsequent coronary revascularization, continuous perioperative 12-lead ST-segment trend monitoring, cTn-I and/or cTn-T, and creatine kinase-MB fraction in the first three postoperative days, were prospectively collected in 501 consecutive elective major vascular procedures. RESULTS: Moderate to severe inducible ischemia on PTS was associated with a 49.0% incidence of low-level (cTn-I >0.6 and/or cTn-T >0.03 ng/ml) and 22.4% conventional (cTn-I >1.5 and/or cTn-T >0.1 ng/ml) troponin elevation. In contrast, patients with preoperative coronary revascularization had 23.4% and 6.4% low-level and conventional troponin elevations, respectively, similar to patients without ischemia on PTS. By multivariate logistic regression, ischemia on PTS was the most important predictor of both low-level and conventional troponin elevations (adjusted odds ratios [ORs] 2.5 and 2.7, p = 0.02 and 0.04, respectively), whereas preoperative coronary revascularization predicted less troponin elevations (adjusted ORs 0.35 and 0.16, p = 0.045 and 0.022, respectively). Postoperative ischemia (>10 min), the more so prolonged (>30 min) ischemia was the only independent predictor of troponin elevation if added with the preoperative predictors to the multivariate analysis (ORs 15.8 and 22.8, respectively, p < 0.001). CONCLUSIONS: Troponin elevations occur frequently after vascular surgery. They are strongly associated with postoperative ischemia, predicted by inducible ischemia on PTS, and reduced by preoperative coronary revascularization. PMID- 15358023 TI - Ischemia, revascularization, and perioperative troponin elevation after vascular surgery. PMID- 15358024 TI - A dietary and exercise intervention slows menopause-associated progression of subclinical atherosclerosis as measured by intima-media thickness of the carotid arteries. AB - OBJECTIVES: The object of this study was to assess the effects of menopause and a diet/exercise intervention on subclinical atherosclerosis progression. BACKGROUND: Subclinical atherosclerosis has been linked to higher coronary heart disease and stroke rates and is greater among postmenopausal women according to cross-sectional analyses. Whether menopause is associated with an accelerated progression of subclinical disease is unknown, as is the extent to which lifestyle intervention can alter the course of progression. METHODS: Intima-media thickness (IMT) measures of the common carotid artery (CCA), internal carotid artery (ICA), and bulb segments of the carotid arteries were measured twice during the course of 4 years in 353 women from the Women's Healthy Lifestyle Project, a dietary and exercise clinical trial designed to prevent adverse risk factor changes through the menopause. A third measure was obtained 2.5 years later for 113 women. RESULTS: The progression of IMT was observed for the average of all segments (AVG), the CCA, and the bulb (0.007 mm/year, 0.008 mm/year, and 0.012 mm/year; p < 0.01 for all), but not for the ICA. Among controls, menopause was associated with accelerated IMT progression (0.003 mm/year for premenopausal women vs. 0.008 mm/year for perimenopausal/postmenopausal women for AVG IMT; p = 0.049). Additionally, among the 160 perimenopausal/postmenopausal women, the intervention slowed IMT progression (0.008 mm/year for the control group vs. 0.004 mm/year for the intervention group for AVG IMT; p = 0.02). Similar results were found for the CCA and bulb segments. CONCLUSIONS: These data demonstrate that the menopause transition is associated with accelerated subclinical atherosclerosis progression and that a diet/exercise intervention slows menopause related atherosclerosis progression. PMID- 15358025 TI - Diet and exercise for perimenopausal women lifestyle interventions can decrease cardiovascular risk. PMID- 15358026 TI - Sublingual nitroglycerin used in routine tilt testing provokes a cardiac output mediated vasovagal response. AB - OBJECTIVES: We set out to determine the effect of sublingual nitroglycerin (NTG), as used during routine tilt testing in patients with unexplained syncope, on hemodynamic characteristics and baroreflex control of heart rate (HR) and systemic vascular resistance (SVR). BACKGROUND: Nitroglycerin is used in tilt testing to elicit a vasovagal response. It is known to induce venous dilation and enhance pooling. Also, NTG is lipophilic and readily passes cell membranes, and animal studies suggest a sympatho-inhibitory effect of NTG on circulatory control. METHODS: Routine tilt testing was conducted in 39 patients with suspected vasovagal syncope (age 36 +/- 16 years, 18 females). Patients were otherwise healthy and free of medication. Before a loss of consciousness set in, oncoming syncope was cut short by tilt-back or counter-maneuvers. Finger arterial pressure was monitored continuously (Finapres). Left ventricular stroke volume (SV) was computed from the pressure pulsations (Modelflow). Spontaneous baroreflex control of HR was estimated in the time and frequency domains. RESULTS: During tilt testing, 22 patients developed presyncope. After NTG administration but before presyncope, SV and cardiac output (CO) decreased (p < 0.001), whereas SVR and HR increased (p < 0.001) in all patients. Arterial pressure was initially maintained. Baroreflex sensitivity decreased after NTG. On Cox regression analysis, the occurrence of a vasovagal response was related to a drop in SV after NTG (hazard ratio 0.86, p = 0.005). CONCLUSIONS: The cardiovascular response to NTG is similar in vasovagal and non-vasovagal patients, but more pronounced in those with tilt-positive results. The NTG facilitated presyncope appears to be CO-mediated, and there is no evidence of NTG induced sympathetic inhibition. PMID- 15358027 TI - Differences in mechanisms and outcomes of syncope in patients with coronary disease or idiopathic left ventricular dysfunction as assessed by electrophysiologic testing. AB - OBJECTIVES: This study evaluated the causes of syncope and the significance and differences in left ventricular (LV) dysfunction, coronary disease, and idiopathic dilated cardiomyopathy (DCM). BACKGROUND: Risk stratification of and indications for an automated defibrillator could differ according to the cause of LV dysfunction. METHODS: Electrophysiologic study, including atrial and ventricular programmed stimulation, was performed in 119 patients with coronary disease (group I) and 61 patients with DCM (group II) with an left ventricular ejection fraction (LVEF) <40% and syncope. Patients were followed from one to six years (mean 4 +/- 2 years). RESULTS: Sustained monomorphic ventricular tachycardia (VT) was induced in 44 group I patients (37%) and 13 group II patients (21%); ventricular flutter (>270 beats/min) or ventricular fibrillation (VF) was induced in 24 group I patients (19%) and 9 group II patients (15%); and various other arrhythmias were identified. Syncope remained unexplained in 34 group I patients (30%) and 16 group II patients (27%). Prognosis depended on the heart disease: VT or VF induction was a predictive factor of mortality in coronary disease and identified a group with high cardiac mortality (46%), compared with patients with a negative study, who had a lower mortality (6%; p < 0.001) than in other studies. Cardiac mortality was only correlated with LVEF in DCM. CONCLUSIONS: Various causes could explain syncope in 70% of patients with coronary disease and DCM, but differences were noted: VT was frequent in coronary disease with a bad prognosis, and ischemia could explain syncope; in DCM, different causes such as atrial tachycardia could be responsible for syncope, but the prognosis only depended on LVEF. PMID- 15358028 TI - Comprehensive analysis of the beta-myosin heavy chain gene in 389 unrelated patients with hypertrophic cardiomyopathy. AB - OBJECTIVES: We sought to determine the prevalence and phenotype of beta-myosin heavy chain gene MYH7 mutations in a large cohort of unrelated patients with hypertrophic cardiomyopathy (HCM). BACKGROUND: Hypertrophic cardiomyopathy is a heterogeneous cardiac disease. MYH7 mutations are one of the most common genetic causes of HCM and have been associated with severe hypertrophy, young age of diagnosis, and high risk of sudden cardiac death. However, these clinical findings from large, family studies have not been confirmed in a large unrelated cohort. METHODS: Deoxyribonucleic (DNA) samples obtained from 389 HCM outpatients seen at this tertiary referral center were analyzed for mutations, using polymerase chain reaction, denaturing high-performance liquid chromatography, and DNA sequencing for all 38 protein-coding exons of MYH7. Clinical data were extracted from patient records blinded to patient genotype. RESULTS: Fifty-eight patients (15%) harbored 40 different mutations in MYH7. Compared with HCM patients without MYH7 mutations, HCM patients with MYH7 were younger at diagnosis (32.9 vs. 42.7 years, p = 0.0002), had more hypertrophy (left ventricular wall thickness of 24.2 vs. 21.1 mm, p = 0.0009), and more frequently underwent myectomy (60% vs. 38%, p = 0.002). The HCM patients with MYH7 mutations more often had a family history of HCM (43% vs. 29%, p = 0.006), but there was no difference in family history of sudden death (16% vs. 14%, p = NS). CONCLUSIONS: In this setting, HCM patients with MYH7 were diagnosed at a younger age and had more hypertrophy, but they had no greater frequency of sudden death among first degree relatives. Although these associations may prove useful for targeted gene screening, caution should be exercised in terms of using pathogenic status in risk stratification. PMID- 15358029 TI - Age and gender affect ventricular-vascular coupling during aerobic exercise. AB - OBJECTIVES: The goal of this study was to examine the age-associated differences in ventricular-vascular coupling, defined by the ratio of arterial elastance (EaI) to left ventricular systolic elastance (E(LV)I), and its components, at rest and during exercise. BACKGROUND: Ejection fraction (EF) increases during exercise, but the EF reserve decreases with aging. Ejection fraction is inversely related to EaI/E(LV)I, an index of the interaction between arterial and ventricular properties, which is an important determinant of cardiac performance. Thus, age differences in EaI/E(LV)I during exercise, due to age differences in EaI, E(LV)I, or both, may help to explain the age deficit in EF reserve. METHODS: We noninvasively characterized EaI/E(LV)I = end-systolic volume index (ESVI)/stroke volume index (SVI) and its two determinants EaI = end-systolic pressure/SVI, and E(LV)I = end-systolic pressure/ESVI, at rest and during exercise in 239 healthy men and women (age range, 21 to 87 years). Blood pressures were assessed with cuff sphygomanometry, and cardiac volumes with gated blood pool scintingraphy. RESULTS: Resting EaI/E(LV)I was not age related in men or women. In both sexes, EaI/E(LV)I decreased during exercise and declined to a lesser extent in older subjects. There were gender differences in the components of EaI/E(LV)I during exercise: EaI was greater in older versus young women (p = 0.01) but was unaffected by age in men. Left ventricular systolic elastance increased to a greater extent in young versus older subjects (p = 0.0001 for men, p = 0.07 for women). CONCLUSIONS: Age-associated differences in EaI/E(LV)I occur in both genders during exercise. Sub-optimal ventricular-vascular coupling helps to explain the age-associated blunting of maximal exercise EF, and its underlying mechanisms appear to differ between men and women. PMID- 15358030 TI - Subclavian artery stenosis: prevalence, risk factors, and association with cardiovascular diseases. AB - OBJECTIVES: The objective was to assess the prevalence of subclavian artery stenosis (SS) in four cohorts (two free-living and two clinical populations) and determine both risk factors for this condition and the association with other cardiovascular conditions. BACKGROUND: The prevalence of SS in the general population is unknown, and its association with risk factors and other cardiovascular diseases is not well-established. METHODS: A total of 4,223 subjects (2,975 from two free-living cohorts and 1,248 from two clinical cohorts) were included in this cross-sectional analysis. Subclavian artery stenosis was defined as > or =15 mm Hg interarm pressure difference. RESULTS: The prevalence of SS was 1.9% in the free-living cohorts and 7.1% in the clinical cohorts; SS was significantly (p < 0.05) associated with past smoking (odds ratio [OR] = 1.80), current smoking (OR = 2.61), and higher levels of systolic blood pressure (OR = 1.90 per 20 mm Hg). Higher levels of high-density lipoprotein (HDL) cholesterol were inversely and significantly associated with SS (OR = 0.87 per 10 mg/dl). In regression analyses relating SS to other cardiovascular diseases, the only significant finding was with peripheral arterial disease (PAD) (OR = 5.11, p < 0.001). CONCLUSIONS: Significant SS is present in approximately 2% of the free living population and 7% of the clinical population. Additionally, SS is correlated with current and past smoking histories, systolic blood pressure, HDL levels (inversely), and the presence of PAD. These findings suggest that bilateral brachial blood pressure measurements should routinely be performed in patients with an elevated risk profile, both to screen for SS, and to avoid missing a hypertension or PAD diagnosis because of unilateral pressure measurement in an obstructed arm. PMID- 15358031 TI - Inhibition of vascular oxidative stress in hypercholesterolemia by eccentric isosorbide mononitrate. AB - OBJECTIVES: We sought to determine if the nitric oxide (NO) donor isosorbide mononitrate (ISMN) (200 mg/kg body weight/day) decreases vascular bioavailability of superoxide in atherosclerosis. BACKGROUND: Vascular oxidative stress limits the bioavailability of endothelial NO and promotes atherosclerosis, while NO itself exerts antioxidative effects. It is unknown if therapeutic NO impacts on vascular oxidative stress in atherosclerosis. METHODS: New Zealand white rabbits (n = 10 each group) were fed either normal chow (control), cholesterol chow (CHOL) (0.75%), or cholesterol chow enriched with ISMN (CHOL-ISMN). Rabbits were fed twice daily. After 16 weeks we used aortic segments to measure vascular superoxide (5-microM lucigenin), intimal lesion formation, and vasoreactivity to acetylcholine (ACH) and ISMN. RESULTS: Plasma cholesterol increased by 40-fold in CHOL and CHOL-ISMN. The plasma concentration of ISMN in CHOL-ISMN was 1,529 +/- 447 ng/ml. Superoxide formation (control: 228 +/- 20 counts/20 min/mg) was strongly enhanced in CHOL (345 +/- 46 counts/20 min/mg, p = 0.02) but not in CHOL ISMN (229 +/- 23 counts/20 min/mg) demonstrating antioxidative effects of eccentric ISMN in vivo. In parallel, intima-media thickness of thoracic aorta (159 +/- 4 microm in control) was reduced from 645 +/- 41 microm (CHOL) to 440 +/ 51 microm (CHOL-ISMN, p < 0.05). Likewise, eccentric ISMN partially restored vascular responses to the NO donor S-nitroso-N-acetyl-D,L-penicillamine and improved endothelium-dependent vasorelaxation. The maximal ACH relaxation increased from 26.3 +/- 9.6% in CHOL to 49.7 +/- 8.1% in CHOL-ISMN; ISMN treatment induced a moderate nitrate tolerance as evidenced by diminished ISMN induced vasodilation. CONCLUSIONS: These data suggest that eccentric ISMN can completely inhibit the increase of vascular bioavailability of superoxide and partially prevent intimal lesion formation and endothelial dysfunction in hypercholesterolemia. PMID- 15358032 TI - Long-term therapy with organic nitrates: the pros and cons of nitric oxide replacement therapy. PMID- 15358033 TI - Co-administration of nitric oxide-aspirin (NCX-4016) and aspirin prevents platelet and monocyte activation and protects against gastric damage induced by aspirin in humans. AB - OBJECTIVES: The goal of this study was to test the hypothesis that NCX-4016 may have broader anti-inflammatory and antithrombotic effects as well as better gastric tolerability than aspirin in humans. BACKGROUND: NCX-4016 is an aspirin derivative containing a nitric oxide-releasing moiety that prevents platelet activation and modulates tissue factor (TF) expression and cytokine release from lipopolysaccharide (LPS)-stimulated monocytes. METHODS: This was a blind observer, placebo-controlled, parallel-group study in which 48 healthy subjects were randomized to receive NCX-4016 800 mg twice a day, NCX-4016 800 mg twice a day plus aspirin 325 mg, aspirin 325 mg, or placebo for 21 days. RESULTS: Similar to aspirin alone, NCX-4016 effectively inhibited platelet aggregation induced by 0.6 mmol/ arachidonic acid, clot-stimulated thromboxane (TX) B2 generation in whole blood, and urinary excretion of 11-dehydro-TXB2. Unlike aspirin alone, the administration of NCX-4016 significantly inhibited TF expression in monocytes stimulated ex vivo with 10 micromol/l LPS (determined by flow-cytometry analysis of TF on CD14 positive cells). NCX-4016 also inhibited the rapid TF expression induced in monocytes by a proteinase activated receptor agonist (thrombin receptor activator protein, 2 micromol/l) as well as LPS-induced expression of CD11b . Ex vivo, release of MCP-1 and interleukin-6 were significantly inhibited by NCX-4016, but not by aspirin. NCX-4016 was not associated with gastric damage, and significantly reduced gastric injury when co-administered with aspirin, although both drugs reduced gastric PGE2 production to the same extent. CONCLUSIONS: NCX-4016 is equally effective as aspirin in inhibiting cyclooxygenase activity. However, NCX-4016 causes less gastric damage and prevents monocyte activation. Larger multicenter trials are warranted to establish clinical efficacy and safety of NCX-4016. PMID- 15358034 TI - Theriac found? Nitric oxide-aspirin and the search for the universal cure. PMID- 15358035 TI - Treatment of acute myocardial infarction by hepatocyte growth factor gene transfer: the first demonstration of myocardial transfer of a "functional" gene using ultrasonic microbubble destruction. AB - OBJECTIVES: We examined whether ultrasonic microbubble destruction (US/MB) enables therapeutic myocardial gene transfer of hepatocyte growth factor (HGF) for acute myocardial infarction (MI). BACKGROUND: Hepatocyte growth factor gene transfer provides cardioprotective effects in MI, which requires direct intramyocardial injection or special vectors. Although US/MB was used in myocardial gene transfer, its feasibility in transfer of a therapeutic gene with non-viral vector remains unknown. METHODS: In a rat model of acute MI, naked plasmid (pVaxl) encoding human HGF (1,500 microg) was infused into the left ventricular (LV) chamber during US/MB (HGF-US/MB) or insonation only (HGF-US) or alone (HGF-alone), while control MI rats received empty pVaxl during US/MB (pVaxl US/MB). For US/MB, transthoracic intermittent insonation with a diagnostic transducer (1.3 MHz) was performed for 2 min at a peak negative pressure of 2,160 kPa during intravenous 20% Optison. RESULTS: Baseline risk area was comparable among the groups. Immunohistology seven days after treatment revealed significant myocardial expression of HGF protein only in HGF-US/MB. At three weeks, LV weight in HGF-US/MB (0.89 +/- 0.03 g) was significantly lower than those in HGF-alone (1.09 +/- 0.08 g), HGF-US (1.04 +/- 0.07 g), and pVaxl-US/MB (1.04 +/- 0.05 g). Moreover, scar size was significantly smaller (16 +/- 6% vs. 39 +/- 5%, 41 +/- 6%, and 40 +/- 4% of total myocardial circumferential length, respectively), while capillary density (49 +/- 8 vs. 34 +/- 5, 37 +/- 6, and 36 +/- 4 capillaries/high-power field, respectively) and arterial density (37 +/- 7 vs. 15 +/- 9, 18 +/- 4, and 14 +/- 11 arterioles/high-power field, respectively) in the risk area were higher in HGF-US/MB than the other groups. CONCLUSIONS: Ultrasound-mediated microbubble destruction may enable myocardial HGF gene transfer with systemic administration of naked plasmid, which enhances angiogenesis, limits infarction size, and prevents LV remodeling after MI. PMID- 15358036 TI - Injectable fibrin scaffold improves cell transplant survival, reduces infarct expansion, and induces neovasculature formation in ischemic myocardium. AB - OBJECTIVES: In this study, we determined whether fibrin glue improves cell transplant retention and survival, reduces infarct expansion, and induces neovasculature formation. BACKGROUND: Current efforts in restoring the myocardium after myocardial infarction (MI) include the delivery of viable cells to replace necrotic cardiomyocytes. Cellular transplantation techniques are, however, limited by transplanted cell retention and survival within the ischemic tissue. METHODS: The left coronary artery of rats was occluded for 17 min followed by reperfusion. One week later, bovine serum albumin (BSA), fibrin glue, skeletal myoblasts in BSA, or skeletal myoblasts in fibrin glue were injected into the infarcted area of the left ventricle. The animals were euthanized five weeks after injection, and their hearts were excised, fresh frozen, and sectioned for histology and immunohistochemistry. RESULTS: After five weeks, the mean area covered by skeletal myoblasts in fibrin glue was significantly greater than the area covered by myoblasts injected in BSA. Myoblasts within the infarct were often concentrated around arterioles. The infarct scar size and myoblasts in the fibrin group were significantly smaller than those in the control and BSA groups. Fibrin glue also significantly increased the arteriole density in the infarct scar as compared with the control group. CONCLUSIONS: This study indicates that fibrin glue increases cell transplant survival, decreases infarct size, and increases blood flow to ischemic myocardium. Therefore, fibrin glue may have potential as a biomaterial scaffold to improve cellular cardiomyoplasty treat and MIs. PMID- 15358037 TI - Roles of endogenous monocyte chemoattractant protein-1 in ischemia-induced neovascularization. AB - OBJECTIVES: We sought to investigate the role of endogenous monocyte chemoattractant protein (MCP)-1 in ischemia-induced neovascularization. BACKGROUND: Roles of inflammatory changes including macrophage infiltration are suggested in ischemic neovascularization. METHODS: Unilateral hindlimb ischemia was induced by excising surgically the entire femoral artery and vein in mice. Immediately after operation, plasmid deoxyribonucleic acid encoding a dominant negative mutant of MCP-1 (7ND) or the empty plasmid (mock) was injected into the ipsilateral thigh adductor muscle. RESULTS: In mock-treated mice, MCP-1 was upregulated transiently in ischemic hindlimb peaking at day 3. Serial laser Doppler blood flow (LDBF) analysis showed an abrupt decrease in blood flow, followed by a recovery to the near-normal levels in mock-treated mice; 7ND treatment had no effects on the initial decrease in LDBF but deteriorated the recovery. At day 3, macrophage infiltration and inductions of tumor necrosis factor (TNF)-alpha and vascular endothelial growth factor (VEGF) were prominent in the ischemic adductor muscle in mock-treated mice; 7ND treatment significantly reduced macrophage infiltration and suppressed TNF-alpha and VEGF inductions in response to ischemia. At day 21, postmortem angiography and anti-CD31 immunohistostaining revealed well-developed collateral vessels and capillary formation, respectively, in the ischemic muscle of mock-treated mice; 7ND overexpression remarkably suppressed the collateral vessel formation and capillary formation. CONCLUSIONS: Endogenous MCP-1 may play a role in ischemia induced neovascularization by recruiting macrophages that activate TNF-alpha and VEGF inductions. PMID- 15358039 TI - Choice of prosthetic heart valves: 20-year results of the Edinburgh Heart Valve Trial. PMID- 15358041 TI - Increased randomness of heart rate could explain increased heart rate variability preceding onset of atrial fibrillation. PMID- 15358043 TI - Measurement of circulating vascular endothelial growth factor in obese subjects. PMID- 15358045 TI - ACC/AHA guidelines for the management of patients with ST-elevation myocardial infarction--executive summary. A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Writing Committee to revise the 1999 guidelines for the management of patients with acute myocardial infarction). PMID- 15358046 TI - Implications of recent clinical trials for the National Cholesterol Education Program Adult Treatment Panel III Guidelines. AB - The Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program issued an evidence-based set of guidelines on cholesterol management in 2001. Since the publication of ATP III, 5 major clinical trials of statin therapy with clinical end points have been published. These trials addressed issues that were not examined in previous clinical trials of cholesterol-lowering therapy. The present document reviews the results of these recent trials and assesses their implications for cholesterol management. Therapeutic lifestyle changes (TLC) remain an essential modality in clinical management. The trials confirm the benefit of cholesterol-lowering therapy in high-risk patients and support the ATP III treatment goal of low-density lipoprotein cholesterol (LDL-C) <100 mg/dL. They support the inclusion of patients with diabetes in the high-risk category and confirm the benefits of LDL-lowering therapy in these patients. They further confirm that older persons benefit from therapeutic lowering of LDL-C. The major recommendations for modifications to footnote the ATP III treatment algorithm are the following. In high-risk persons, the recommended LDL-C goal is <100 mg/dL, but when risk is very high, an LDL-C goal of <70 mg/dL is a therapeutic option, ie, a reasonable clinical strategy, on the basis of available clinical trial evidence. This therapeutic option extends also to patients at very high risk who have a baseline LDL-C < 100 mg/dL. Moreover, when a high-risk patient has high triglycerides or low high-density lipoprotein cholesterol (HDL-C), consideration can be given to combining a fibrate or nicotinic acid with an LDL-lowering drug. For moderately high-risk persons (2+ risk factors and 10-year risk 10% to 20%), the recommended LDL-C goal is <130 mg/dL, but an LDL-C goal <100 mg/dL is a therapeutic option on the basis of recent trial evidence. The latter option extends also to moderately high-risk persons with a baseline LDL-C of 100 to 129 mg/dL. When LDL-lowering drug therapy is employed in high-risk or moderately high risk persons, it is advised that intensity of therapy be sufficient to achieve at least a 30% to 40% reduction in LDL-C levels. Moreover, any person at high risk or moderately high risk who has lifestyle-related risk factors (eg, obesity, physical inactivity, elevated triglycerides, low HDL-C, or metabolic syndrome) is a candidate for TLC to modify these risk factors regardless of LDL-C level. Finally, for people in lower-risk categories, recent clinical trials do not modify the goals and cutpoints of therapy. PMID- 15358047 TI - ACC/AHA guidelines for the management of patients with ST-elevation myocardial infarction; A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee to Revise the 1999 Guidelines for the Management of patients with acute myocardial infarction). PMID- 15358049 TI - The control of thyroid-specific gene expression: what exactly have we learned as yet? AB - The identification of transcription factors TTF 1 and Pax 8 and the demonstration of their pivotal role in thyroid development and in the control of thyroid specific gene expression, although representing remarkable openings in our understanding of cell-specific transcription in the thyroid, still leave a lot of open questions. Recent work investigating the development of thyroid-specific gene expression in transgenic mouse models, now reveal that some basic assumptions have to be reconsidered also. Altogether, currently available data indicate that the regulatory machinery undergoes significant changes during thyroid organogenesis and confirm the existence of still unknown factors whose roles appear at least as critical as the ones played by TTF 1 and Pax 8 in the control of specific gene expression. PMID- 15358050 TI - The formation of an intrachain disulfide bond in the leptin protein is necessary for efficient leptin secretion. AB - Leptin is a cytokine secreted by the adipose tissue that is involved in the control of body weight. We previously showed that a point mutation (R105W) in leptin results in leptin deficiency, marked obesity and hypogonadism in humans adults. Expression in COS1 cells showed impaired secretion and intracellular accumulation of the mutated protein. However, impaired secretion of the mutant leptin had not been demonstrated in adipose cells. In this work, we demonstrate that secretion of R105W mutant is impaired in rat and human adipocytes. We also show that R105W mutant expressed in COS1 cells and in PAZ6 adipocytes forms large molecular aggregates that cannot cross a filtration membrane with a cut-off of 100 kDa. Moreover, we have engineered, by site directed mutagenesis, the cDNAs coding for leptin in which either Cys 117, Cys 167, or both, were replaced by a serine. When expressed in COS1 cells or PAZ6 adipocytes, cysteine mutants also show impaired secretion and formation of large molecular aggregates. Therefore, our work indicates that the formation of an intramolecular disulfide bridge is necessary for normal processing and secretion of leptin. Moreover, the similarity of the behavior of R105W mutant and cystein mutants suggests that the lack of secretion observed with the naturally occurring mutant could result from impaired disulfide bond formation. PMID- 15358051 TI - Saccharomyces cerevisiae phosphoenolpyruvate carboxykinase: relevance of arginine 70 for catalysis. AB - Saccharomyces cerevisiae phosphoenolpyruvate (PEP) carboxykinase is a key enzyme of the gluconeogenic pathway and catalyzes the decarboxylation of oxaloacetate and transfer of the gamma-phosphoryl group of ATP to yield PEP, ADP, and CO2 in the presence of a divalent metal ion. Previous experiments indicate that mutation of amino acid residues at metal site 1 decrease the enzyme catalytic efficiency and the affinity of the protein for PEP, evidencing the relevance of hydrogen bond interactions between PEP and water molecules of the first coordination sphere of the metal ion for catalysis [Biochemistry 41 (2002) 12763]. To further understand the function of amino acid residues located in the PEP binding site, we have now addressed the catalytic importance of Arg70, whose guanidinium group is close to the PEP carboxyl group. Arg70 mutants of PEP carboxykinase were prepared, and almost unaltered kinetic parameters were found for the Arg70Lys PEP carboxykinase, while a decrease in 4-5 orders of magnitude for the catalytic efficiency was detected for the Arg70Gln and Arg70Met altered enzymes. To evaluate the enzyme interaction with PEP, the phosphopyridoxyl-derivatives of wild type, Arg70Lys, Arg70Gln, and Arg70Met S. cerevisiae PEP carboxykinase were prepared, and the change in the fluorescence emission of the probe upon PEP binding was used to obtain the dissociation equilibrium constant of the corresponding derivatized enzyme-PEP-Mn2+ complex. The titration experiments showed that a loss in 2.1 kcal/mol in PEP binding affinity is produced in the Arg70Met and Arg70Gln mutant enzymes. It is proposed that the electrostatic interaction between the guanidinium group of Arg70 and the carboxyl group of PEP is important for PEP binding and for further steps in catalysis. PMID- 15358052 TI - Characterization of the oligosaccharide component of microsomal beta glucuronidase from rat liver. AB - The oligosaccharides of microsomal beta-glucuronidase were analysed by gel permeation and weak anion exchange chromatography following hydrazine release. N linked glycans, constituted 80% of the total glycan pool and were mainly of the tri- and biantennary complex type with or without core and arm fucose. The major oligosaccharide, that comprised 30.6% of all the species analysed, was structurally identified by reagent array analysis method and found to be a triantennary complex structure, Galbeta1,4GlcNAcbeta1,2Manalpha1,6(3)(Galbeta1,4GlcNAcbeta1,4(Galbeta1,4GlcNAcbet 1,2) Manalpha1,3(6))Manbeta1,4GlcNAcbeta1,4 GlcNAc. O-Linked glycans comprised 20% of the total glycan pool, the major species being Galbeta1,3GalNAc. All of the N- and O-linked glycans were charged. Most of the negative charge was due to sialic acid (85.0%) with the remainder being phosphate present as phosphomonoesters (7.3%) and phosphodiesters (5%). This is the first report of O linked carbohydrate chains in microsomal beta-glucuronidase. The presence of O linked glycans and branched N-linked glycans in a microsomal enzyme, in relation to the current view of glycosyltransferase compartmentalization in the Golgi is discussed. PMID- 15358053 TI - Cell culture media are potent antioxidants that interfere during LDL oxidation experiments. AB - In vitro cell-induced low-density lipoprotein (LDL) oxidation is a model frequently used for studies on antioxidant compounds which may be potentially antiatherogens. Using Cu2+ or the free radical generator 2,2'-azobis-[2 amidinopropane] dihydrochloride (AAPH) to oxidize human LDL, we showed that the cell culture media Ham's F10 and RPMI are potent antioxidants which reduce LDL protective effect of various thyroid compounds. The culture media interfered with the compounds depending on their mechanism of action, and RPMI had the greatest antioxidant effect, completely hiding antioxidant efficiency of the compounds whatever the prooxidant agent was. We suggest some recommendations for study of antioxidant compounds using cell-induced LDL oxidation models. PMID- 15358054 TI - The tryptophane residues of dimeric arginine kinase: roles of Trp-208 and Trp-218 in active site and conformation stability. AB - Roles of the two tryptophane residues of dimeric arginine kinase (AK) were individually investigated by site-directed mutagenesis. Both residues were fully conserved in the phosphogen kinase family and the mutant proteins were analyzed by enzyme kinetics, fluorescence spectroscopy, fluorescence quenching experiments, thermal stability and conformational stability. Our studies revealed that Trp-218 was located at the active site of AK and was the major fluorescence contributor (96.9%). Single replacement of this residue by alanine led to almost complete inactivation of the enzyme. In addition, a decrease in the melting temperature in differential scanning calorimetry (DSC) profiles and the equilibrium studies in guanidine hydrochloride (GdnHCl) denaturation after mutagenesis also suggested that Trp-218 takes part in stabilizing the conformational structure of AK. Although another tryptophane, Trp-208 was not located at the active sites, it may take part in maintaining the correct dimer conformation for catalysis. Replacement of this tryptophane by alanine decreased the activity to 70.3% and made it susceptible to heat and denaturants, such as GdnHCl. In addition, Trp-208 also seemed to play an important role in correct protein folding. PMID- 15358055 TI - Biochemical, genetic and physiological characterization of venom components from two species of scorpions: Centruroides exilicauda Wood and Centruroides sculpturatus Ewing. AB - Current literature concerning the taxonomic names of two possibly distinct species of scorpions from the genus Centruroides (sculpturatus and/or exilicauda) is controversial. This communication reports the results of biochemical, genetic and electrophysiological experiments conducted with C. exilicauda Wood of Baja California (Mexico) and C. sculpturatus Ewing of Arizona (USA). The chromatographic profile fractionation of the soluble venom from both species of scorpions is different. The N-terminal amino acid sequence for nine toxins of C. exilicauda was determined and compared with those from C. sculpturatus. Lethality tests conducted in mice support the idea that C. exilicauda venom should be expected to be medically less important than C. sculpturatus. Thirteen genes from the venomous glands of the scorpion C. exilicauda were obtained and compared with previously published sequences from genes of the species C. sculpturatus. Genes coding for cytochrome oxidase I and II of both species were also sequenced. A phylogenetic tree was generated with this information showing important differences between them. Additionally, the results of electrophysiological assays conducted with the venom from both species on the Ca(2+)-dependent K(+) channels, showed significant differences. These results strongly support the conclusion that C. exilicauda and C. sculpturatus are in fact two distinct species of scorpions. PMID- 15358056 TI - Instability of monoclonal myeloma protein may be identified as susceptibility to penetration and binding by newly synthesized Congo red derivatives. AB - Monoclonal myeloma proteins often have an abnormal, unstable structure, and tend to aggregate with fatal clinical consequences. A method for early clinical identification of this aggregation tendency is impatiently awaited. This work proposes the use of supramolecular dyes as specific ligands to reveal protein instability. Disclosure of excessive polypeptide chain flexibility in unstable monoclonal proteins, leading to increased susceptibility to penetration by foreign compounds, appeared possible when new supramolecular Congo red-derived dyes with different protein-binding capabilities were used for complexation. Two basic protein instability levels, local and global, were differentiated by comparing the extent of protein loading with dye and the subsequent electrophoretic migration rate of the complexes. A simple electrophoretic test is proposed for assessment of the instability of monoclonal proteins in clinical conditions. PMID- 15358057 TI - Metal-ion induced conformational changes in alkaline phosphatase from E. coli assessed by limited proteolysis. AB - Alkaline phosphatase (AP) displays significant structural changes during metal ion binding, supporting cooperative interactions between the subunits of the dimeric enzyme. Here, we present data on the dynamic properties of AP from E. coli, and characterize the structural changes that accompany variations in metal ion content, combining limited proteolysis and MALDI-TOF mass spectrometry. Limited proteolysis revealed an internal cleavage site at Arg-293, reflecting a position of conformational flexibility supporting subunit communication essential for catalysis. A specific shielding of a region distant from the metal-binding site has been demonstrated, implying transmission of conformational changes, induced by metal-ion binding to the adjacent subunit, across the subunit interface. PMID- 15358058 TI - DNA interstrand cross-linking by a mycotoxic diepoxide. AB - The diepoxide mycotoxin (2R, 3R, 8R, 9R)-4,6-decadiyne-2,3:8,9-diepoxy-1,10-diol (repandiol) was both isolated from the mushroom Hydnum repandum and synthesized de novo. Repandiol was found to form interstrand cross-links within a restriction fragment of DNA, linking deoxyguanosines on opposite strands primarily within the 5'-GNC and 5'-GNNC sequences preferred by diepoxyoctane. However, repandiol was a significantly less efficient cross-linker than either of the diepoxyalkanes (diepoxyoctane and diepoxybutane) to which it was compared. PMID- 15358059 TI - The absorption of polarized light by vertebrate photoreceptors. AB - A physiologically realistic model has been constructed for a theoretical study of the mechanisms by which the vertebrate visual system absorbs linearly polarized light. Using a 4 x 4 matrix technique, analytic solutions to Maxwell's equations have been deduced for rod and cone photoreceptors, allowing calculation of the absorbance as a function of wavelength for a variety of illumination geometries. With the use of experimentally measured optical parameters, the calculated absorbance spectra show excellent agreement in both magnitude and form with microspectrophotometric data. Moreover, failing to correct for the true nature of reflection or scattering in the sample, results in the elevated absorbance commonly seen at shorter wavelengths in experimental measurements. Finally, calculated dichroic ratios also accurately predict experimental results, mirroring the differences seen between rods and cones. PMID- 15358060 TI - Illusory motion reversal is caused by rivalry, not by perceptual snapshots of the visual field. AB - In stroboscopic conditions--such as motion pictures--rotating objects may appear to rotate in the reverse direction due to under-sampling (aliasing). A seemingly similar phenomenon occurs in constant sunlight, which has been taken as evidence that the visual system processes discrete "snapshots" of the outside world. But if snapshots are indeed taken of the visual field, then when a rotating drum appears to transiently reverse direction, its mirror image should always appeared to reverse direction simultaneously. Contrary to this hypothesis, we found that when observers watched a rotating drum and its mirror image, almost all illusory motion reversals occurred for only one image at a time. This result indicates that the motion reversal illusion cannot be explained by snapshots of the visual field. The same result is found when the two images are presented within one visual hemifield, further ruling out the possibility that discrete sampling of the visual field occurs separately in each hemisphere. The frequency distribution of illusory reversal durations approximates a gamma distribution, suggesting perceptual rivalry as a better explanation for illusory motion reversal. After adaptation of motion detectors coding for the correct direction, the activity of motion-sensitive neurons coding for motion in the reverse direction may intermittently become dominant and drive the perception of motion. PMID- 15358061 TI - Denoising and contrast constancy. AB - Contrast constancy is the ability to perceive object contrast independent of size or spatial frequency, even though these affect both retinal contrast and detectability. Like other perceptual constancies, it is evidence that the visual system infers the stable properties of objects from the changing properties of retinal images. Here it is shown that perceived contrast is based on an optimal thresholding estimator of object contrast, that is identical to the VisuShrink estimator used in wavelet denoising. PMID- 15358062 TI - Axotomized mouse retinal ganglion cells containing melanopsin show enhanced survival, but not enhanced axon regrowth into a peripheral nerve graft. AB - Melanopsin is found in only approximately 2% of mouse retinal ganglion cells (RGCs), making these RGCs uniquely and directly photosensitive. Given that the majority of RGCs die after axotomy and that grafting of a peripheral nerve to the eye provides a permissive environment for axon regrowth, the present study examined the survival and axonal regrowth of melanopsin-containing RGCs in mice. One month after optic nerve transection and grafting, RGCs with regrown axons were labeled from the grafts and retinae were processed to visualize melanopsin and TUJ1. Melanopsin-positive and negative RGCs were counted and compared to axotomized RGCs from ungrafted eyes and uninjured RGCs. Melanopsin-positive RGCs showed a 3-fold increase in survival rate compared to non-melanopsin RGCs. Despite this enhanced survival, melanopsin-containing RGCs did not show increased axon regrowth into nerve grafts. PMID- 15358063 TI - Characteristics of saccadic intrusions. AB - Primary fixation is never perfectly stable, but is frequently interrupted by slow drifts, microsaccades and saccadic intrusions (SI). SI are involuntary, conjugate movements which take the form of an initial fast movement away from the desired eye position and followed after a short duration, by either a return secondary saccade or a drift. The purpose of this study was to examine the prevalence and metrics of SI in a population of 50 healthy subjects. Using both one and two dimensional recordings we find that all 50 members of the subject group exhibited SI. The SI were bilateral, conjugate and horizontal. No purely vertical SI were detected when examined in three subjects. SI amplitude mean and range was 0.6 degrees +/-0.5 degrees, 0.1 degrees -4.1 degrees; SI frequency mean and range was 18.0+/-14.3 per min, 1.0-54.8 per min; SI duration mean and range was 225+/-150, 20-870 ms. The mean SI amplitude and frequency when SI<0.5 degrees were removed was 0.97 degrees +/-0.56 degrees and 7.0+/-11.4 per min respectively. Age was positively correlated with SI amplitude (p<0.01), but there was no correlation between age and SI frequency. Three of four types of SI monophasic square wave intrusions (MSWI), biphasic square wave intrusions (BSWI) and double saccadic pulses (DSP) were found to be exclusively saccadic, whilst the fourth type, the single saccadic pulses (SSP), were confirmed to exhibit a slow secondary component. MSWI were the most frequently observed SI occurring in 47 out of 50 (94%) of the subjects with a mean amplitude, frequency and duration of 0.7 degrees +/-0.5 degrees, 11.5+/-11.6 per min, and 255+/-147 ms respectively. Mean amplitudes and frequencies for BSWI (n=20), SSP (n=11) and DSP (n=34) were found to be 0.50 degrees +/-0.2 degrees, 1.2+/-2.5 per min; 0.40 degrees +/-0.20 degrees, 0.4+/-1.0 per min and 0.3 degrees +/-0.4 degrees, 5.0+/-8.7 per min respectively. No differences in MSWI characteristics were found between binocular and monocular viewing. Possible explanations for SI occurrence include experimental viewing conditions, subject fatigue and covert shifts in attention. PMID- 15358064 TI - Development of a viewing strategy during adaptation to an artificial central scotoma. AB - Although many individuals with a central scotoma develop eccentric fixation most often beneath or left of the scotoma, little is known about how they come to develop a particular viewing strategy. We investigated this by asking eight subjects with normal vision to read isolated letters, words and text passages while an artificial scotoma covered a central portion of the visual field. We quantified viewing strategy and analysed changes in their viewing behaviour over 8-10 sessions within a two-week period. Subjects read while either a horizontal (n=4) or vertical bar scotoma (n=4), 10 degrees wide, covered the entire horizontal or vertical meridian of the stimulus field. For the horizontal scotoma group: (1) there was an increasing preference to use the inferior visual field for isolated letters/words and text passages, which was essentially complete within the test period; (2) the superior visual field was preferred when reading letters/words initially presented in upper visual space and the inferior visual field when reading letters/words initially presented in lower visual space; (3) in general, variation in viewing strategy according to stimulus position diminished over the sessions for all stimuli. For the vertical scotoma group: (1) two subjects used the left and right visual fields in approximately equal proportion to view isolated letters/words, one subject showed a weak preference to use the left visual field and one subject developed a strong preference for using the right visual field; (2) the text passages could be read with combined use of left and right visual fields in a specific manner; (3) the left visual field was preferred to view stimuli initially presented in left visual space while the right visual field was preferred for words initially presented in right visual space. This effect diminished across sessions. Overall, these findings indicate that (1) a specific viewing strategy can be developed through as little as 5 hours of reading experience without guided training; (2) two distinctly separate retinal areas can be used in an integrated manner during reading; (4) stimulus position in visual space can influence viewing strategy; (5) in general, reading encourages a preference for the inferior over the superior visual field, but not the left over right visual field. Letter/word/text recognition and reading speeds increased progressively across sessions, even after scotoma lateralisation appeared stabilised suggesting that multiple mechanism are involved in adaptive changes. PMID- 15358065 TI - Programming of double-step saccade sequences: modulation by cognitive control. AB - The capacity to detect and correct errors is thought to engage cognitive control. To probe the nature of such control in relation to eye movements, subjects performed a double-step task under different instructions: to FOLLOW the appearance of successive targets; or to cancel the initial saccade and REDIRECT gaze to the final target location. Saccade sequences occurred in the FOLLOW and REDIRECT conditions where they represented correct and corrective behaviour, respectively. We observed that corrective responses were faster than correct responses, and concurrent preparation of saccades was facilitated during error correction. These results are consistent with psychological theories that posit supervisory cognitive control over action during error correction. PMID- 15358066 TI - Orientation discrimination across the visual field: matching perceived contrast near threshold. AB - Performance can often be made equal across the visual field by scaling peripherally presented stimuli according to F=1+E/E2 where E2 is the eccentricity at which stimulus size must double to maintain foveal performance levels. Previous studies suggest that E2 for orientation discrimination is in the range of 1.5 degrees -2 degrees when stimuli are presented at contrasts well above detection threshold. Recent psychophysical and physiological evidence suggests spatial reorganization of receptive fields at near-threshold contrasts. Such contrast-dependent changes in receptive field structure might alter the amount of size scaling necessary to equate task performance across the visual field. To examine this question we measured orientation discrimination thresholds for a range of stimulus sizes and eccentricities (0 degrees -15 degrees ). We used the same procedure previously employed except that stimuli were presented at near threshold contrasts. We controlled for the effects of perceptual contrast on thresholds through a matching procedure. A standard line of 3 degrees in length presented at fixation was set to 2 just noticeable differences above detection threshold. The perceived contrast of all other stimuli was adjusted by the subject to match this one. Orientation discrimination thresholds were then obtained at these matching contrasts for all stimulus sizes and eccentricities. E2 values of 3.42 degrees and 3.50 degrees were recovered for two subjects; these values were about a factor of two larger than E2 values previously found for this task when stimuli were presented at higher physical contrasts. PMID- 15358067 TI - Oculomotor control in asymptomatic and recently diagnosed individuals with the genetic marker for Huntington's disease. AB - We compared oculomotor control among individuals in the early stages of Huntington's disease (HD), with that of individuals who are presymptomatic HD gene carriers (PSGC) and nongene carriers (NGC). The oculomotor testing paradigm included both traditional tests and a novel experimental procedure to assess visual scanning. Traditional tests elicited saccades, pursuit and optokinetic nystagmus (OKN). HD patients demonstrated marked delay in the initiation of volitional saccades (anti-saccade and memory-guided saccades), a reduced number of correct volitional saccades, reduced velocity of saccades, and a decreased OKN gain. We also studied visual scanning while the participants completed the Digit Symbol Subscale of the Wechsler Adult Intelligence Survey-Revised (WAIS-R). The HD participants demonstrated an abnormal gaze strategy, which may be associated with attention and/or planning deficits. Differences between the PSGC and NGC groups were only observed for two measures: PSGC had a decreased number of memory guided saccades and a subtle delay in the initiation of volitional saccades. Our results suggest that oculomotor measures are a sensitive biomarker in the early stage of HD and demonstrate that the combination of more traditional oculomotor tests with visual scanning tests is useful in the evaluation of visual performance. PMID- 15358068 TI - Influence of age, speed and duration of monotonous driving task in traffic on the driver's useful visual field. AB - Recent research has shown that the useful visual field deteriorates in simulated car driving when the latter can induce a decrease in the level of activation. The first aim of this study was to verify if the same phenomenon occurs when driving is performed in a simulated road traffic situation. The second aim was to discover if this field also deteriorates as a function of the driver's age and of the vehicle's speed. Nine young drivers (from 22 to 34 years) and nine older drivers (from 46 to 59 years) followed a vehicle in road traffic during two two hour sessions. The car-following task involved driving at 90 km.h(-1) (speed limit on road in France) in one session and at 130 km.h(-1) (speed limit on motorway in France) in the other session. While following the vehicle, the driver had to detect the changes in colour of a luminous signal located in the central part of his/her visual field and a visual signal that appeared at different eccentricities on the rear lights of the vehicles in the traffic. The analysis of the data indicates that the useful visual field deteriorates with the prolongation of the monotonous simulated driving task, with the driver's age and with the vehicle's speed. The results are discussed in terms of general interference and tunnel vision. PMID- 15358069 TI - Components of motion perception revealed: two different after-effects from a single moving object. AB - If motion that one has been looking at for some time suddenly stops, or if one shifts one's gaze to a static object, one will see motion in the opposite direction: the motion after-effect. If two transparent surfaces move with different speeds in different directions, then the direction of the motion after effect will depend on the test pattern. For such transparent surfaces both the local motion and the global percept have two components. When looking at a normal moving object, there is only one perceived global motion. However, we know that locally there can be considerable ambiguity (the aperture problem). Does one adapt to all the local components, including those that one does not perceive, or only to the perceived global motion? We designed a stimulus that is perceived to be a fast rotating object, but also has a slow local radial component of motion. By selecting an appropriate test pattern we could either get a radial or a rotating motion after-effect. Thus we show that adaptation to motion must (also) occur at a stage at which local motions have not yet been integrated to give a unified percept. PMID- 15358070 TI - The effect of disrupting the human magnocellular pathway on global motion perception. AB - The purpose of this study was to demonstrate the effect of human magnocellular (M)-pathway disruption on global motion perception. Coherence thresholds for global motion direction discrimination in random dot patterns were determined at slow and moderate dot speeds: (1) after adaptation to full-field sinusoidal flicker or a steady gray field, and (2) on a red or a gray background. Adaptation to flicker and a red background increased motion coherence thresholds relative to the gray baseline conditions at both dot speeds. Physiological studies have shown that M cells in the retina and LGN are inhibited by red light and are a main contributor to flicker perception in monkeys. Therefore, our results suggest that interference with processing in the subcortical M pathway disrupts higher-level motion integration. PMID- 15358071 TI - Preservation of vision following cell-based therapies in a model of retinal degenerative disease. AB - This study examines whether subretinal transplantation of a human retinal pigment epithelial cell line (ARPE19) or human Schwann cells early in the course of degeneration can limit the loss of visual acuity that normally occurs in dystrophic Royal College of Surgeons (RCS) rats as they age. The acuity of both transplanted groups was significantly better than controls at all ages tested, though the rescue profile of each cell type was unique. These data indicate that the transplantation of cells with very different phenotypes can be used to limit the deterioration of spatial vision in an animal model of retinal degenerative disease. PMID- 15358072 TI - Cross-orientation summation in texture segregation. AB - Human texture vision has been modeled as a filter-rectify-filter (FRF) process, in which '2nd-order' filters detect changes in the rectified outputs of luminance based '1st-order' filters. This study tested the validity of the two basic assumptions of the standard FRF model, namely (a) that the 2nd-order filters are sensitive to spatial modulations in both contrast and orientation, and (b) that the 2nd-order filters are tuned to different 1st-order orientations. In the first experiment, we tested subthreshold summation between two orthogonal carrier orientations in detection of a texture region, which was defined by contrast modulations across regions in the two carrier orientations, while systematically varying the relative change magnitudes between the two orientations. The results showed that the detection thresholds were determined by spatial difference in the contrast integrated over the two orientations. Orientation difference did act as a segregation cue, but only when there was no differences in carrier contrast. This suggests that two mechanisms are involved in texture segregation; one that detects changes in luminance contrast and another that detects changes in orientation. To further analyze the latter mechanism, a second experiment measured cross-orientation summation in the detection of purely orientation defined textures, using stimuli that were density modulations of two orientations presented among randomly-orientated distractors. Again, the relative modulation magnitudes between the two orientations was systematically varied. The results are consistent with the notions that (a) the dominant orientation is extracted from the 1st-order outputs before the 2nd-order process, and that (b) the 2nd order, spatial comparison process integrates those dominant signals over different orientations. PMID- 15358073 TI - Factors limiting suprathreshold vision measured by a flash-sound simultaneity paradigm. AB - Internal noise and sampling efficiency are the main factors which limit visual performance. In a previous study [Vis. Res. 43 (2003) 1103] we compared the variance of human reaction time to that of an ideal observer and found that the sampling efficiency to suprathreshold stimuli was much lower than that obtained in detection experiments. In order to bypass the effects of the motor system on visual performance, we used a flash-sound simultaneity paradigm. We found that the sampling efficiency for 0.4- and 4-c/deg near-threshold Gabor patches is higher only by a factor of 2.5 than that to above-threshold patterns. The signal dependent multiplicative internal noise was similar to the additive internal noise at lower signal contrast levels and exceeded it at higher signal contrast levels. The results show that real observers' performance for detecting suprathreshold stimuli can be accounted for by a model taking into account the non-linear visual-signal transduction and multiplicative components of the internal noise induced by the signal and external noise. In addition, this model assumes that performance depends on the response duration, rather than signal duration. The results imply that the multiplicative internal noise induced by high contrast visual signals determines performance for suprathreshold visual detection. PMID- 15358074 TI - Backward masking and the central performance drop. AB - Kehrer [Spatial Vision 2 (1987) 247] found that texture discrimination performance sometimes peaks in the parafovea rather than at the fovea, and he referred to this phenomenon as the central performance drop (CPD). Kehrer used a backward mask to limit performance and Morikawa [Vision Res. 40 (2000) 3517] argued that in some cases the temporal aspects of the backward mask may be critical to the emergence of the CPD. In one experiment Morikawa showed that the CPD does not emerge when a simultaneous noise-mask (different from the mask used by Kehrer) is used to limit performance. In another experiment Morikawa showed that unmasked texture displays comprising short lines do not elicit the CPD. In both cases, changes in the temporal aspects of the texture displays were accompanied by changes in the spatial structure of the mask or stimulus. For the spatio-temporal theory of the CPD to be sustained one would have to show that noise masks elicit a CPD when used as backward masks and that the short-line textures elicit a CPD when followed by backward masks. Our evidence provides little if any support for either of these predictions. Furthermore, an analysis of a simple filter-rectify-filter model of texture segmentation shows that a greatly attenuated CPD is to be expected when a noise mask is used as a source of spatial noise. PMID- 15358075 TI - The influence of spatial frequency and contrast on saccade latencies. AB - We characterised the impact of spatial frequency and contrast on saccade latencies to single Gabor patches. Saccade latencies decreased as a function of contrast, and increased with spatial frequency. The observed latency variations are qualitatively similar to those observed for manual reaction times. For single target detection, our findings highlight the similarity in the visual processes that support both saccadic and manual responses. PMID- 15358076 TI - Stopping the motion and sleuthing the flash-lag effect: spatial uncertainty is the key to perceptual mislocalization. AB - A moving object is perceived to lie beyond a static object presented at the same time at the same retinal location (flash-lag effect or FLE). Some studies report that if the moving stimulus stops moving (flash-terminated condition or FTC) the instant the flash occurs, a FLE does not occur. Other studies, using different stimuli, report that the FLE does, in fact, occur in the FTC. The FTC is thus a crucial turning point in theories of flash-lag. Unraveling the mystery of the FLE in the FTC will help unravel the mechanisms underpinning flash-lag and perhaps even perceptual localization in general. Our experiments show that eccentricity of the moving stimulus was a contributing factor, as were eccentricity of the flashed stimulus and spatial separation between the two stimuli. Other factors, such as contrast and offset of moving stimulus, also modulate the magnitude of the FLE in the FTC. We surmise that uncertainty in determining the position in space of a moving stimulus is a key requirement for the lag-effect. A lag-effect in the FTC challenges influential models, such as differential latency, motion extrapolation, and postdiction. Based partly on the notion of an asymmetric spread of activity that arises because of the sheer nature of motion and from a combination of established physiological mechanisms, we propose a schematic account of the present findings that subsumes previous psychological models and scaffolds past experimental findings. PMID- 15358077 TI - Asymmetry between horizontal and vertical illusory lines in determining the depth of their embedded surface. AB - To investigate how the visual system integrates disparity information from horizontal and vertical edges and conveys it to the regions without any depth cues, we introduce a new phenomenon of subjective surface formation in an Ehrenstein-style configuration with inducing elements at different depths, and without explicit monocular occlusion zones. Different sets of experiments by separate groups of subjects suggest that when a subjective (illusory) square forms, it is at the depth of vertical illusory sides rather than horizontal ones. When the vertical side inducers are stereoscopically behind the horizontal ones, subjective surface formation is less likely. In depth assignment, we interpret the dominance of vertical sides over horizontal ones geometrically: vertical orientation can convey the horizontal disparity--a critical factor for Wheatstone (classic) stereopsis--but horizontal orientation per se lacks horizontal disparity information. Therefore, in the disparity integration, vertical illusory sides play a dominant role and their depth information influences the embedded subjective surface as well as the horizontal illusory sides. PMID- 15358078 TI - Glass pattern studies of local and global processing of contrast variations. AB - Using Glass patterns [Nature 223 (1969) 578; Nature 246 (1973) 360; Perception 5 (1976) 67], we have studied the role of contrast differences in local and global processes of form perception. The virtue of these patterns (composed of a set of randomly distributed elements combined with a geometrically transformed copy) for studying object formation is that they allow ready isolation of local processes, the combination of dots to form a perceptual pair, from global processes, the combination of dipoles into the percept of an overall rotational or translational pattern. We find that a contrast difference within dot-pairs reduces the ability to resolve local features; large differences totally abolish the perception of the pattern. Contrast differences between dot-pairs lessen, but do not abolish, the global integration among local features. In both cases the effect is proportional to the ratio of the two contrast levels employed. Effects which differ for rotations and translations, are consistent with the greater areal integration required to resolve rotational patterns. PMID- 15358079 TI - Structural determinants of the PDE6 GAF A domain for binding the inhibitory gamma subunit and noncatalytic cGMP. AB - Photoreceptor cGMP phosphodiesterases (PDE6 family) are modular enzymes with each catalytic subunit containing two N-terminal regulatory GAF domains, GAF A and GAF B. The GAF A domains contribute to dimerization of the PDE6 catalytic subunits and to binding of the inhibitory Pgamma subunits, and represent candidate sites for noncatalytic binding of cGMP. We performed a mutational analysis of selected residues from the GAF A domain of cone PDEalpha' to identify the cGMP-binding pocket and delineate the Pgamma-binding surface. Results of this analysis establish the noncatalytic cGMP-binding site within the PDE6 GAF A domain and suggest that occupation of the pocket by cGMP is required for high-affinity binding of Pgamma to the proximate contact surface. PMID- 15358080 TI - In vivo biometry in the mouse eye with low coherence interferometry. AB - PURPOSE: A major drawback of the mouse model of myopia is that the ocular dimensions cannot be measured in vivo, and that histological techniques post mortem suffer from limited resolution. We have tested the potential of a newly developed technique, optical low coherence interferometry (OLCI), adapted for short measurement distances by Meditec, Carl Zeiss, Jena, Germany (the "ACMaster"). Using this technique, ocular biometry was performed in mice with normal vision and after deprivation of form vision. METHODS: Axial eye length, corneal thickness and anterior chamber depth were measured in 23 mice, aged 25-53 days, and standard deviations from repeated measurements in the same eyes, as well as intra-individual and inter-individual variability were determined in different age groups. The data were compared to those from a preceding study in which biometrical data were obtained from frozen sections [Vision Res. 44 (2004) 1857]. Refractions were measured by automated infrared photorefraction. Mice had either normal visual exposure or were monocularly deprived of form vision for 14 days. RESULTS: Using OLCI, axial length could be determined with an average standard deviation of 8.0 +/- 2.9 microm, corneal thickness with 3.5 +/- 2.1 microm, and anterior chamber depth with 10.6 +/- 12.3 microm. Neither axial length, nor corneal thickness, nor anterior chamber depth were significantly different in left and right eyes of individual mice that had normal visual experience (mean absolute difference between axial lengths: 17 +/- 18 microm, between corneal thickness 5.1 +/- 4.8 microm, and between anterior chamber depths 16.7 +/- 14.8 microm). Compared to the variability that was previously found in frozen sections, the variability of axial length measurements with OLCI was 2.7 times less. After two weeks of form deprivation, OLCI revealed a significant axial elongation in the occluded eyes, compared to the contralateral fellow eyes (+38 +/- 36 microm or 1.16%, p = 0.045, n = 7, paired t-test). In this sample, no accompanying myopic shift was observed in the occluded eyes but this observation is not unexpected given the inherently variable responses of mouse eye growth to visual deprivation. CONCLUSION: OLCI had sufficient resolution in living mice to detect axial length changes in vivo that were equivalent to a dioptric change of 2 D. Using this technique, it was confirmed that mouse eyes respond to form deprivation by axial elongation, similar to the eyes of other animal models. The lack of a myopic shift in this sample, despite the axial elongation, demonstrates that biometric data are particularly important when the mouse eye is used as a model to study myopia. PMID- 15358081 TI - Center-surround antagonism in spatial vision: retinal or cortical locus? AB - Mach and Hering had early advanced a model of spatial visual processing featuring an antagonistic interaction between adjoining areas in the visual field. Spatial opponency was one of the first findings when single-unit studies of the retina were begun. Not long afterwards psychophysical experiments revealed a center surround organization closely matching that found in the mammalian retina. It hinged on the demonstration of reduction of sensitivity in a small patch of the visual field when its surround was changed from dark to bright. Because such patterns inevitably produce borders, well-known phenomena of border interaction could be seen as providing alternative explanations, whose substrate would most likely be in the visual cortex. These competing viewpoints are discussed especially as they pertain to the recent demonstration of spatial differences in the center/surround organization between the normal and affected eyes of amblyopes. To the extent that most findings favor a retinal site for the psychophysically measured antagonism, and that evidence is accumulating for a direct effect on the mammalian retina of stimulus manipulation during visual development, the difference in spatial parameters of center/surround antagonism in amblyopia suggests that the dysfunction in amblyopia begins already in the retina. PMID- 15358082 TI - Contribution of rod and cone pathways to the dark-adapted electroretinogram (ERG) b-wave following retinal degeneration in RCS rats. AB - Although the RCS rat is widely used as a model of progressive photoreceptor loss, it is unclear how the relative rod and cone functions change with age. Rod and cone b-waves were isolated using a double flash ERG paradigm. In contrast to cones, rods never reached normal functional maturity levels, and the ERG b-wave changed from being predominantly rod-driven to being purely cone-driven by age 74 days, at which point, b-waves were progressively replaced by negative STR-like (scotopic threshold response) waves that persisted up to age 180 days. A double flash commonly abolished this wave and unveiled a b-wave. PMID- 15358083 TI - Perception of mirror symmetry in amblyopic vision. AB - Mirror symmetry is ubiquitous in natural visual scenes, and detection of mirror symmetry seems to be a global, automatic, effortless and important aspect of visual perception. The perception of mirror symmetry has not been studied in humans with amblyopia. In this paper we measured and quantified the detection of mirror symmetry in adults with naturally occurring amblyopia. Our results show that amblyopia may severely impair the detection of mirror symmetry, and that this impairment is not simply a consequence of reduced stimulus visibility. Rather, we suggest that this loss may reflect, at least in part, a deficit in the integration of local orientation information. PMID- 15358084 TI - A unified theory of brightness contrast and assimilation incorporating oriented multiscale spatial filtering and contrast normalization. AB - Brightness induction includes both contrast and assimilations effects. Brightness contrast occurs when the brightness of a test region shifts away from the brightness of adjacent regions. Brightness assimilation refers to the opposite situation in which the brightness of the test region shifts toward that of the surrounding regions. Interestingly, in the White effect [Perception 8 (1979) 413] the direction of the induced brightness change does not correlate with the amount of black or white border in contact with the gray test patch. This has led some investigators to reject spatial filtering explanations not only for the White effect but for brightness perception in general. Instead, these investigators have offered explanations based on a variety of junction analyses and/or perceptual organization schemes. Here, these approaches are challenged with a critical set of new psychophysical measurements that determined the magnitude of the White effect, the shifted White effect [Perception 10 (1981) 215] and the checkerboard illusion [R.L. DeValois, K.K. DeValois, Spatial Vision, Oxford University Press, NY, 1988] as a function of inducing pattern spatial frequency and test patch height. The oriented difference-of-Gaussians (ODOG) computational model of Blakeslee and McCourt [Vision Res. 39 (1999) 4361] parsimoniously accounts for the psychophysical data, and illustrates that mechanisms based on junction analysis or perceptual inference are not required to explain them. According to the ODOG model, brightness induction results from linear spatial filtering with an incomplete basis set (the finite array of spatial filters in the human visual system). In addition, orientation selectivity of the filters and contrast normalization across orientation channels are critical for explaining some brightness effects, such as the White effect. PMID- 15358085 TI - Effect of background motion on the perceived shape of a 3-D object. AB - The effect of the speed of a background surface on the judged shape of a moving object was investigated in four experiments. Observers judged the magnitude of a concave dihedral angle translating or rotating against a planar background. Judged angle magnitude decreased (indicating an increase in perceived depth) with increasing background speed until the background speed reached the speed of the front edge of the angle. Judged angle magnitude then increased with background speed until the difference between the background and front edge speed was large. A model that was previously proposed to account for angle magnitude judgments from translations and rotations is extended to displays with a moving background. PMID- 15358086 TI - Internal surface representations approximated by reverse correlation. AB - We presented two nai ve observers with 20,000 random-dot stereograms. On each trial, the observers had to indicate the presence or absence of a complex 3D pattern (a large '+' sign in relief). However, unbeknownst to them, the stereograms did not contain any signal, but only disparity noise. Responses and verbal reports indicate that the observers 'saw' the suggested 3D surface configuration in roughly half the trials even though structured local low-level signal was never presented. Using reverse correlation, we derived an approximation of the internal surface-based representations, or templates, that best accounted for the observers' responses. These templates were shown to be spatially well defined and temporally stable. We propose that the 3D surface based representations that we derived are the first approximations and depictions of the intermediary process that allows the visual system to successfully link degraded, bottom-up signal and high-level, top-down object recognition. PMID- 15358087 TI - Children with dyslexia: evidence for visual attention deficits in perception of rapid sequences of objects. AB - The attentional blink (AB) refers to a decrease in accuracy that occurs when observers are required to identify, detect or classify the second of two rapidly sequential targets. The AB is typically attributed to an inability to rapidly reallocate attentional resources from the first to the second target. Thus, it provides an ideal tool to investigate how visual attention is rapidly allocated to sequences of stimuli such as occurs when reading. In the present work, we compared the magnitude of the AB in children with developmental dyslexia to reading-matched and age-matched control groups. In Experiment 1, when two targets were presented in the same spatial location, the AB deficit was similar in the reading-matched and dyslexic groups, but greater in the dyslexic group than in age-matched controls. In Experiment 2, when targets were presented in different spatial locations, performance in the dyslexic group was worse than the age matched controls and marginally worse than the reading-matched controls. Taken together, the results argue for developmental delays in the ability of children with dyslexia to allocate attention to rapidly-sequential stimuli, as well as some evidence for difficulties that are unique to this group. PMID- 15358088 TI - Metacontrast masking is specific to luminance polarity. AB - A 1 degrees -spot was flashed up on a screen, followed by a snugly fitting annular mask. We measured the amount of masking as a function of stimulus luminance. The surround was always mid-gray, the masking ring was either black or white, and the luminance of the spot target ranged from 0% to 100% of white in 4% steps. Observers reported the apparent lightness of the masked spot by adjusting a matching spot. RESULTS: A black annular mask made all spots that were darker than the gray surround appear to be transparent, that is, of the same luminance as the surround (complete masking). The black ring had virtually no masking effect on spots that were lighter than the surround. Conversely, a white ring made all spots that were lighter than the gray surround look apparently the same luminance as the surround (complete masking), but had virtually no masking effect on spots that were darker than the surround. In summary, a black ring masked spatial decrements but not increments, whilst a white ring masked spatial increments but not decrements. Thus masking occurred only when the spot and the ring had the same luminance polarity. This same-polarity masking still occurred when the target spot was larger than the 'donut hole' of the masking ring, so that the target and ring partly overlapped. This ruled out simple edge cancellation theories. Instead, masking disrupts the filling-in process that normally propagates inward from the edges of a spot [Vision Res. 31 (7-8) (1991) 1221]. We conclude that metacontrast masking occurs within, but not between, separate visual ON and OFF pathways. PMID- 15358089 TI - Regiospecific alkaline protease-catalyzed divinyl acyl transesterifications of primary hydroxyl groups of mono- and di-saccharides in pyridine. AB - This paper describes highly selective transesterification reactions, catalyzed by an alkaline protease from Bacillus subtilis in pyridine, of several mono- and di saccharides with divinyl dicarboxylates ranging from 4 to 10 carbon atoms. A series of polymerizable vinyl fatty acid sugar esters were obtained with good selectivity and high yields. Most products had high proportions of the alpha anomer. The influences of the enzymes, solvents, temperature, and acyl donor chain length on the reaction were studied. Vinyl sugar esters offer a new family of functional water-soluble monomers for preparation of sugar-containing polymers. PMID- 15358090 TI - Crystal and molecular structure and absolute configuration of lincomycin hydrochloride monohydrate. AB - Lincomycin is a broad-spectrum antibiotic synthesized by Streptomyces lincolnensis that is particularly active against Gram-positive bacteria. It is widely used in human and veterinary applications. The crystal structure of lincomycin has been undertaken with a view to obtain the conformational and structural features of the drug in order to afford a comparison of its structural features with other aminoglycoside antibiotics. We report here the details of its structural and conformational features as determined by single-crystal X-ray crystallography. Crystals of lincomycin hydrochloride are orthorhombic, space group P2(1)2(1)2, with the cell dimensions a=18.5294(3) Angstroms, b=20.5980(4) Angstroms, c=6.17380(10) Angstroms, V=2356.35(7) Angstroms3. The structure was solved using X-ray diffraction data and refined to a final R-value of 0.0391 for 2321 reflections (I > or = 2sigma). The absolute configuration was established using the anomalous dispersion of the sulfur and chlorine atoms in the structure. The molecule consists of an amino acid linked by an amide group to a monosaccharide of galactose stereochemistry. A network of hydrogen-bonds stabilizes the crystal structure. PMID- 15358091 TI - Mitochondrial proteins in neuronal degeneration. AB - In this review, we highlight recent findings about the role of some mitochondrial proteins in neurological diseases. Studies in mice gene-deleted for Omi/HtrA2 and AIF showed the involvement of these mitochondrial proteins in selective cell degeneration in the spinal cord and brain. In humans, mutations in the mitochondrial protein, Paraplegin, cause an autosomal form of hereditary spastic paraplegia with an enhanced sensitivity to oxidative stress. Reactive oxygen species and decreased respiratory chain activity in mitochondria also contribute to common neurological diseases. The mitochondrial uncoupling protein, Ucp-2, was found to be neuroprotective in experimental stroke and brain trauma. Recent proteomic and profiling studies have revealed the existence of additional mitochondrial proteins with unknown functions. The elucidation of the physiological functions of mitochondrial proteins may lead to new insights into the role of these organelles in cell degeneration and to identification of novel drug targets for the prevention and treatment of different diseases. PMID- 15358092 TI - Protection against collagen-induced arthritis by electrotransfer of an expression plasmid for the interleukin-4. AB - Rheumatoid arthritis (RA) is a chronic inflammatory joint disease, leading to cartilage and bone destruction. We investigated whether the electrotransfer of IL 4 DNA could regulate the disease progress of murine collagen-induced arthritis (CIA). The maximum serum level of mIL-4 was measured by 340 pg/ml on day 1 following DNA transfer. The onset of severe CIA and the degree of synovitis and cartilage erosion were significantly reduced in mice treated with IL-4 DNA (P<0.05). The beneficial effect of IL-4 gene transfer lasted for at least 17 days subsequent to treatment. The expression of IL-1beta was considerably decreased in the paws by IL-4 DNA transfer (P<0.01). On the contrary, the ratio of TIMP2 to MMP2 significantly increased in the IL-4 DNA-treated group (P<0.01). These data demonstrated that electroporation-mediated gene transfer could provide a new approach as an IL-4 therapy for autoimmune arthritis. PMID- 15358093 TI - Stimulation of lipogenesis in rat adipocytes by ATP, a ligand for P2-receptors. AB - The activation of P2-receptors has a wide range of diverse effects in many tissues. Here we show that extracellular ATP stimulates lipogenesis in adipocytes derived from the epididymal fat pads of male Wistar rats. The lipogenic effect of ATP is not susceptible to treatment of adipocytes with adenosine deaminase or an adenosine receptor antagonist. Degradation of ATP in adipocyte suspension by ectonucleotidases is slow and remaining ATP concentrations are sufficient to activate P2-receptors. ATP does not affect basal or insulin stimulated glucose transport, or basal or isoproterenol stimulated lipolysis, respectively. The lipogenic effect of ATP is mimicked by the adenine compounds, ADP, AMP, and beta,gamma-methylene-ATP, but not by other nucleotides (UTP, UDP, CTP, GTP, ITP, and diadenosine tetraphosphate), indicating that extracellular nucleotides stimulate lipogenesis via a P2-receptor. ATP and its receptor may define a signalling system in adipocytes, which regulates fat stores independently from established hormones. PMID- 15358094 TI - Metabolism of A-ring diastereomers of 1alpha,25-dihydroxyvitamin D3 by CYP24A1. AB - The metabolism of 1alpha,25(OH)(2)D(3) (1alpha,3beta) and its A-ring diastereomers, 1beta,25(OH)(2)D(3) (1beta,3beta), 1alpha,25(OH)(2)-3-epi-D(3) (1alpha,3alpha), and 1beta,25(OH)(2)-3-epi-D(3) (1beta,3alpha), was examined to compare the substrate specificity and reaction specificity of CYP24A1 between humans and rats. The ratio between C-23 and C-24 oxidation pathways in human CYP24A1-dependent metabolism of (1alpha,3alpha) and (1beta,3alpha) was 1:1, although the ratio for (1alpha,3beta) and (1beta,3beta) was 1:4. These results indicate that the orientation of the hydroxyl group at the C-3 position determines the ratio between C-23 and C-24 oxidation pathways. A remarkable increase of metabolites in the C-23 oxidation pathway was also observed in rat CYP24A1-dependent metabolism. The binding affinity of human CYP24A1 for A-ring diastereomers was (1alpha,3beta)>(1alpha,3alpha)>(1beta,3beta)>(1beta,3alpha), indicating that both hydroxyl groups at C-1 and C-3 positions significantly affect substrate-binding. The information obtained in this study is quite useful for understanding substrate recognition of CYP24A1 and designing new vitamin D analogs. PMID- 15358095 TI - Effect of a hyaluronan synthase suppressor, 4-methylumbelliferone, on B16F-10 melanoma cell adhesion and locomotion. AB - Hyaluronan (HA) is a ubiquitous, major component of the extracellular matrix. It is involved in cell adhesion and locomotion, and hence in tumor metastasis. We have previously reported that 4-methylumbelliferone (MU) inhibits HA synthesis and may be a useful tool for examining the functions of HA. We here demonstrate that the formation of cell surface HA by melanoma cells and its release into the culture medium are inhibited by MU. Adhesion and locomotion assays revealed that the adhesion and locomotion of melanoma cells were dose-dependently inhibited by MU. Conversely, treatment with exogenous HA enhanced both adhesion and locomotion. Thus, preventing the formation of cell surface HA reduced both the adhesion and locomotion of melanoma cells, suggesting that MU may act as an inhibitor of tumor metastasis. PMID- 15358096 TI - Human microvascular endothelial cells immortalized with human telomerase catalytic protein: a model for the study of in vitro angiogenesis. AB - Human microvascular endothelial cell-1 (HMEC-1) generated by transfection with SV40 large T antigen has been the prevailing model for in vitro studies on endothelium. However, the transduction of SV40 may lead to unwanted cell behaviors which are absent in primary cells. Thus, establishing a new microvascular endothelial cell line, which is capable of maintaining inherent features of primary endothelial cells, appears to be extremely important. Here, we immortalized primary human microvascular endothelial cells (pHMECs) by engineering the human telomerase catalytic protein (hTERT) into the cells. Endothelial cell-specific markers were examined and the angiogenic responses were characterized in these cells (termed as HMVECs, for human microvascular endothelial cells). We found that VEGF receptor 2 (Flk-1/KDR), tie1, and tie2 expression is preserved in HMVEC, whereas Flk-1/KDR is absent in HMEC-1. In addition, HMVEC showed similar angiogenic responses to VEGF as HMEC-1. Furthermore, the HMVEC line was found to generate a prominent angiogenic response to periostin, a potent angiogenic factor identified recently. The data indicate that HMVEC may serve as a suitable in vitro endothelium model. PMID- 15358097 TI - Identification of extra- and intracellular alanyl aminopeptidases as new targets to modulate keratinocyte growth and differentiation. AB - Aminopeptidase inhibitors strongly affect proliferation, differentiation, and function of immune cells and show therapeutic potential in inflammatory disorders. In psoriatic lesions, keratinocytes display increased cellular turnover and disturbed differentiation, leading to epidermal hyperplasia accompanied by the loss of stratum granulosum. Here, we report in the HaCaT hyperproliferative keratinocyte cell line as well as in two primary keratinocyte strains in vitro a molecular and biochemical analysis of the expression of both membrane and cytosol alanyl aminopeptidase (cAAP) on the mRNA, protein, and enzymatic activity level. We found a clear dose-dependent suppression of DNA synthesis in vitro in the presence of the inhibitors actinonin, bestatin, and the cAAP-specific inhibitor PAC-22 correlating well with the simultaneous decrease in enzyme activity. In vivo, actinonin dose-dependently restored the stratum granulosum and ameliorated the impaired keratinocyte differentiation in the mouse tail model of psoriasis. Taken together, these data suggest that targeting alanyl aminopeptidases may be beneficial for psoriasis and other inflammatory skin disorders. PMID- 15358098 TI - RAMPs and CRLR expressions in osteoblastic cells after dexamethasone treatment. AB - Adrenomedullin (ADM) is a potent stimulator of osteoblastic activity and promotes bone growth in vivo. ADM receptors are formed by heterodimerization of the CRLR and a RAMP2 or RAMP3 molecule. Since glucocorticoid responsive elements were recently identified in the human CRLR promoter and that glucocorticoids exert a major action in bones, we investigated the acute effect of dexamethasone (Dex) treatment on ADM receptor components in osteoblastic cell types: the MC3T3-E1 cells and calvaria-derived osteoblastic cells. Changes in expression of CRLR and RAMPs molecules were evaluated at mRNA levels using RT-PCR and at protein levels by Western blot analysis. We found that Dex increased expression of RAMP1 and RAMP2 mRNA in a time-dependent but dose-independent manner, while RAMP3 was unchanged. In contrast, Dex decreased the CRLR mRNA expression and these changes were reflected at protein levels. We suggest that Dex, in osteoblastic cells, altered ADM receptor by inhibition of CRLR expression and consequently could impair the ADM anabolic effect on bone. Our findings could explain in part, the detrimental side effects observed at bone level during glucocorticoid therapy. PMID- 15358099 TI - A novel RGDS-analog inhibits angiogenesis in vitro and in vivo. AB - In this study the anti-angiogenic action of a novel non-peptide RGDS-analog named RAM was tested in vitro and in vivo. RAM inhibited FGF-2-induced chemotaxis by 80% in an adhesion-independent way. Further, it induced HUVEC-apoptosis in collagen-seeded HUVEC, indicating that such pro-apoptotic effect was adhesion independent. In vivo studies revealed that RAM inhibited FGF-2 induced angiogenesis by 60% in the mouse Matrigel-assay and in the chicken-egg chorion allantoic membrane assay. Finally, RAM was markedly more stable in serum as compared to the template RGDS and after 24 h incubation in 100% serum was significantly more active than RGDS. Taken together these results show that RAM exerts anti-chemotactic and pro-apoptotic effects, by an unexpected adhesion independent mechanism, as we have recently shown for the template RGDS molecule [Blood 103 (2004) 4180], and has in vivo relevant anti-angiogenic properties, with marked stability in serum; therefore, RAM represents a novel promising anti angiogenic molecule. PMID- 15358100 TI - Inhibition of nucleotide excision repair by anti-XPA monoclonal antibodies which interfere with binding to RPA, ERCC1, and TFIIH. AB - The xeroderma pigmentosum group A protein (XPA) binds to three nucleotide excision repair (NER) factors: RPA, ERCC1, and TFIIH. XPA also binds preferentially to UV- or chemical carcinogen-damaged DNA. In this study, we prepared anti-XPA monoclonal antibodies and examined their effects on NER. Two clones inhibited cell-free NER reactions. The mode of inhibition appeared to differ; one clone inhibited both 5' and 3' incisions equally while the other inhibited the 5' incision more. The two clones inhibited the binding of XPA to RPA, ERCC1, and TFIIH. They did not inhibit the binding to damaged DNA either. These results suggest that the interaction of XPA with these NER factors is essential to the NER pathway. The epitopes of these antibodies were located outside of the binding regions for these NER factors. Steric hindrance or conformational changes of XPA brought about by the binding of anti-XPA IgG possibly cause the inhibitory effects. PMID- 15358101 TI - Phosphoinositide-dependent kinase-1 orthologues from five eukaryotes are activated by the hydrophobic motif in AGC kinases. AB - Phosphoinositide-dependent kinase-1 (PDK1) mediates activation of many AGC kinases by docking onto a phosphorylated hydrophobic motif located C-terminal of the catalytic domain in the AGC kinase. The interaction shifts PDK1 into a conformation with increased catalytic activity and leads to autophosphorylation of PDK1. We demonstrate here that addition of a hydrophobic motif peptide increases the catalytic activity of PDK1 orthologues from Homo sapiens, Aplysia californica, Arabidopsis thaliana, Schizosaccharomyces pombe (ksg1), and Saccharomyces cerevisiae (Pkh1 and Pkh2) 2- to 12-fold. Furthermore, the hydrophobic motif peptide increases autophosphorylation of PDK1 from Homo sapiens, S. pombe, and S. cerevisiae (Phk2). Our results suggest that PDK1 interaction and activation by the hydrophobic motif of AGC kinases is a central mechanism in PDK1 function, which is conserved during eukaryotic evolution. PMID- 15358102 TI - STAT3 activation in macrophages following infection with Salmonella. AB - The induction of signal transducer and activators of transcription (STATs) in macrophages is necessary for cellular activation, and we investigated the activation of STAT3 in these cells following infection with Salmonella. Increased activation of STAT3 was observed at 6 and 24 h post-infection in the mesenteric lymph nodes and spleens when compared to control mice. CD11b+ cells isolated from the mesenteric lymph nodes of infected mice demonstrated increased STAT3 activation as early as 6 h following infection. Culturing bone marrow-derived macrophages with Salmonella resulted in translocation of STAT3 to the nucleus and STAT3 phosphorylation as early as 30 min post-exposure. Increased STAT3 activation was also observed in the lymphoid organs or in macrophages from mice deficient for IL-6 or IL-10 production following infection. Taken together, these studies clearly demonstrate an early increase in the activation of STAT3 in vivo and in vitro following infection with wild type Salmonella. PMID- 15358103 TI - Identifying Plasmodium falciparum EBA-175 homologue sequences that specifically bind to human erythrocytes. AB - Erythrocyte binding antigen-160 (EBA-160) protein is a Plasmodium falciparum antigen homologue from the erythrocyte binding protein family (EBP). It has been shown that the EBP family plays a role in parasite binding to the erythrocyte surface. The EBA-160 sequence has been chemically synthesised in seventy 20-mer sequential peptides covering the entire 3D7 protein strain, each of which was tested in erythrocyte binding assays to identify possible EBA-160 functional regions. Five EBA-160 high activity binding peptides (HABPs) specifically binding to erythrocytes with high affinity were identified. Dissociation constants lay between 200 and 460 nM and Hill coefficients between 1.5 and 2.3. Erythrocyte membrane protein binding peptide cross-linking assays using SDS-PAGE showed that these peptides bound specifically to 12, 28, and 44 kDa erythrocyte membrane proteins. The nature of these receptor sites was studied in peptide binding assays using enzyme-treated erythrocytes. HABPs were able to block merozoite in vitro invasion of erythrocytes. HABPs' potential as anti-malarial vaccine candidates is also discussed. PMID- 15358104 TI - Thioredoxin-1 mediates osteoclast stimulation by reactive oxygen species. AB - We found that the antioxidant protein thioredoxin-1 (Trx) is more highly expressed in osteoclasts than in macrophages. Moreover, transfection of RAW 264.7 (RAW) cells with a Trx-expression construct resulted in a dramatic increase in their capacity for osteoclast formation. In contrast, Trx-expression was suppressed and osteoclast formation was abrogated by transfection with the antioxidant proteins glutathione peroxidase-1 (Gpx) or peroxiredoxin-1 (Prx). These divergent effects suggest that Trx augments osteoclast formation through some special function. It is known that Trx enhances the binding of several transcription factors to DNA. We found that AP-1, NFkappaB, and NFAT-reporter gene expression was enhanced more greatly by RANKL in RAW cells transfected with the Trx-expression construct. Thus, oxidants stimulate osteoclastic differentiation by induction of Trx-expression, which augments the DNA binding of transcription factors essential for osteoclastic differentiation. Conversely, antioxidants, including Gpx and Prx, suppress Trx-expression and thereby osteoclastic differentiation. PMID- 15358105 TI - Hypermethylation of the wild-type ferrochelatase allele is closely associated with severe liver complication in a family with erythropoietic protoporphyria. AB - Erythropoietic protoporphyria (EPP) is an inherited disorder of heme biosynthesis caused by cellular decreases in ferrochelatase (FECH) activity. Clinical expression of this disorder usually requires coinheritance of a mutant FECH allele and a normal FECH allele expressed at a low level. In this study, we investigated the methylation status of a normal, but poorly expressed, FECH gene in a single Japanese family with EPP. In this family, the proband died from liver failure, whereas the mother and sister exhibited overt EPP with mild liver dysfunction. A splicing mutation (IVS9+1g-->a) in the FECH gene, which produces a mutant FECH transcript lacking exon 9, was detected in the maternal allele of the proband and his sister. All subjects, including the father, who did not exhibit EPP, possessed the IVS3-48c/c genotype. This allele increases the proportion of aberrantly spliced mRNA, resulting in reduced FECH activity. Normal FECH transcripts were, however, detected in the mother and sister, but not in the proband. The CpG sites in the region from bases -78 to -31 were partially methylated in the proband and his father, but not in his mother or sister. Additionally, CpG methylation within this region reduced transcription of the FECH gene. These results suggest that whereas the combination of a maternal IVS9+1a allele and a paternal IVS3-48c allele results in overt EPP, CpG methylation of the FECH gene promoter, likely inherited from the father, increases the severity of EPP, leading to fatal liver failure, as seen in the proband. PMID- 15358106 TI - Characterization of adrenomedullin in non-human primates. AB - Adrenomedullin (AM) is a 52 amino acid peptide involved in the pathophysiology of several human diseases. Here we show the gene structure, organ distribution, and regulated expression of AM in monkey. The monkey AM (mAM) gene is located on the short arm of chromosome 9 and it codes for a 185 amino acid preprohormone, which contains two amidated peptides identical to the human AM and proadrenomedullin N terminal 20 peptide. The promoter region of the mAM gene contains a variety of transcription factor binding motifs. mAM is widely expressed throughout many organs as shown by real-time PCR and immunohistochemical techniques, and we have found similar levels of circulating plasma AM in monkeys and humans. A significant upregulation of the mAM mRNA was observed in monkey cells exposed to low oxygen tension conditions, TGF-beta1, all-trans-retinoic acid, and dexamethasone. Our collective data show a high degree of homology between mAM and hAM, which renders the monkey an attractive animal model for future pharmacological and pre-clinical studies targeting AM. PMID- 15358107 TI - Human SULT1A3 pharmacogenetics: gene duplication and functional genomic studies. AB - Sulfotransferase (SULT) 1A3 catalyzes the sulfate conjugation of catecholamines. Inheritance is an important factor responsible for individual variation in SULT1A3 activity, and gene resequencing studies have shown the presence of one functionally significant SULT1A3 nonsynonymous cSNP. However, following completion of the Human Genome Project, it appeared that SULT1A3 might be duplicated. We used specific PCR-based assays and fluorescence in situ hybridization to verify that 2 SULT1A3 genes-SULT1A3 and SULT1A4-were present on chromosome 16 in all human DNA samples studied. Furthermore, reanalysis of previous gene resequencing data confirmed the presence of the SULT1A3 SNPs identified previously, but also revealed 11 novel polymorphisms, including 3 nonsynonymous cSNPs. Functional genomic studies showed that two of those cSNPs, C302T, and C302A, resulted in decreased enzyme activity without striking changes in substrate kinetics but with parallel changes in levels of immunoreactive protein. In addition, RT-PCR revealed that both SULT1A3 and SULT1A4 can be transcriptionally active. The duplication of SULT1A3 will have to be taken into account in future efforts to understand individual variation in SULT1A3 activity or properties. PMID- 15358108 TI - Activation of nuclear factor-kappaB and its proinflammatory mediator cascade in the infarcted rat heart. AB - Nuclear transcription factor (NF)-kappaB regulates inflammatory and immune responses by increasing the expression of specific inflammatory genes in various tissues. Whether the infarcted heart includes the activation of NF-kappaB and a proinflammatory mediator cascade that it regulates has not been fully explored. Herein, we monitored the temporal and spatial activation of NF-kappaB, together with expression of tumor necrosis factor (TNF)-alpha, vascular cell adhesion molecule-1 (VCAM-1), and transforming growth factor (TGF)-beta(1), in the infarcted rat heart at and remote to MI from day 3 to day 28 following left coronary artery ligation. Compared to the normal heart, we observed NF-kappaB activation, together with the elevated expression of VCAM-1 in endothelial cells, TNF-alpha and TGF-beta(1) in inflammatory cells at sites of repair in the infarcted heart, which started on day 3, peaked on day 7, and gradually declined thereafter. Our findings suggest NF-kappaB activation and its proinflammatory mediator cascade are contributory to the inflammatory response and remodeling that appear at various sites of repair in the infarcted rat heart. PMID- 15358110 TI - PCR expression mutagenesis: a high-throughput mutation assay applied to the glucocorticoid receptor ligand-binding domain. AB - Glucocorticoid receptors (GRs) are extensively studied members of the steroid hormone receptor superfamily that regulate the transcription rates of numerous genes. Notwithstanding, the role of each GR amino acid in the various steps of transactivation is still unknown. A recent report shows that linear DNA has the same capacity as super-helical plasmid DNA for gene expression in transient transfection assays. Based on this observation, we describe a high-throughput assay to analyze a large set of alanine point mutations that are introduced by two rounds of PCR. The PCR products are then directly transfected into cells. This PCR expression mutagenesis (PEM) technique is used to identify several new residues of the GR ligand binding domain that influence ligand binding and/or transactivation. PEM thus provides a quick method for screening large quantities of mutant proteins. In combination with automation, PEM provides a more rapid and efficient tool for probing the role of each amino acid in the biological functions of a given protein. PMID- 15358109 TI - Alteration of the timing of implantation by in vivo gene transfer: delay of implantation by suppression of nuclear factor kappaB activity and partial rescue by leukemia inhibitory factor. AB - Nuclear factor kappaB (NF-kappaB) is activated in the murine endometrium during implantation period [Am. J. Reprod. Immunol. 51 (2004) 16]. Transient transfection of IkappaBalpha mutant (IkappaBalphaM) cDNA into the mouse uterine cavity using hemagglutinating virus of Japan envelope vector suppressed uterine NF-kappaB activity less than half of that observed in control on days 3.5 and 4.5 p.c. IkappaBalphaM cDNA transfection led to significant delay of implantation. After IkappaBalphaM cDNA transfection, LIF mRNA expression in the uterus was significantly suppressed on days 3.5 and 4.5 p.c. Co-transfection of LIF cDNA with IkappaBalphaM cDNA in the uterus partially rescued the delay of implantation induced by suppression of NF-kappaB activity. Taken together, these findings indicate that NF-kappaB activation determines the timing of the implantation, at least in part, via control of LIF expression. PMID- 15358111 TI - Yeast cytochrome c is a sequence-specific DNA-binding protein. AB - A protein binding to the alcohol oxidase 2 upstream activation sequence (AOX2UAS) of the methylotropic yeast, Pichia pastoris, has been purified and identified as cytochrome c (cyt c). Cyt c purified from P. pastoris or Saccharomyces cerevisiae binds to AOX2UAS. Specific point mutations in AOX2UAS abolish cyt c binding. We conclude that yeast cyt c is a sequence-specific DNA-binding protein and may have a regulatory role in the nucleus. PMID- 15358112 TI - The bortezomib-induced mitochondrial damage is mediated by accumulation of active protein kinase C-delta. AB - Bortezomib (PS-341) is an inhibitor of the S26 proteasome. Bortezomib induces mitochondrial damage but the exact mechanism remains unclear. We studied PKC delta, a kinase that is regulated by proteasome degradation and translocates to mitochondria in apoptosis, and examined whether PKC-delta could be a potential mediator of bortezomib-induced mitochondrial damage. Co-incubation of bortezomib with a PKC-delta inhibitor, rottlerin, suppressed bortezomib-induced apoptosis in U937 cells. Western analysis of U937 cells treated with bortezomib revealed accumulation of full-length PKC-delta in the first 4 h. By 16 h an active catalytic fragment of PKC-delta accumulated in mitochondria. The cleavage of PKC delta after bortezomib treatment was mediated by caspases, because a pan-caspase inhibitor BAF prevented the appearance of the active fragment of PKC-delta. These findings indicate that accumulation of the active PKC-delta fragment in mitochondria is responsible for bortezomib-induced mitochondrial damage. PMID- 15358113 TI - Loss of CYP3A7 gene induction by 1,25-dihydroxyvitamin D3 is caused by less binding of VDR to the proximal ER6 in CYP3A7 gene. AB - Cytochrome P450 3A4 and 3A7 (CYP3A4 and CYP3A7, respectively) are predominant forms in the human adult and fetal liver, respectively. 1,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) is known to be a potent inducer of CYP3A4 in human colon carcinoma Caco-2 via vitamin D receptor (VDR). However, whether CYP3A7 is inducible by 1,25(OH)(2)D(3) has not yet been elucidated. In the present study, we examined the effect of 1,25(OH)(2)D(3) on CYP3A7 gene expression in Caco-2 cells, which express CYP3A4 and CYP3A7 mRNAs. 1,25(OH)(2)D(3) hardly induced the expression of CYP3A7 mRNA in contrast to the marked induction of CYP3A4 mRNA. Reporter assay using 5'-franking region CYP3A4 and CYP3A7 genes also revealed that 1,25(OH)(2)D(3) activates CYP3A4 promoter, but not CYP3A7 promoter, which has two mutations in the proximal ER6 site compared with CYP3A4 promoter. In addition, we found that the binding of VDR to the proximal ER6 in CYP3A7 gene was markedly less than that to the proximal ER6 in CYP3A4 gene using gel shift assay. Taken together, the decrease of VDR binding to the proximal ER6 caused by the mutation results in the loss of CYP3A7 gene activation by 1,25(OH)(2)D(3). PMID- 15358114 TI - Both leptin and leptin-receptor are essential for apolipoprotein M expression in vivo. AB - It was previously shown that circulating levels of leptin and apolipoprotein M (apoM) correlate to each other. In this study, we examined whether plasma leptin and leptin-receptors are of importance for apoM expression in vivo. It was found that in both liver and kidney, expression of apoM was significantly lower in leptin deficient ob/ob mice and in leptin-receptor deficient db/db mice than in control mice. Furthermore, leptin administration (0.5 or 1.5 microg/g body weight) significantly increased plasma apoM levels and apoM mRNA levels in liver and in kidney in ob/ob mice. We conclude that both leptin and leptin-receptor are essential for the apoM expression, indicating that leptin is physiologically regulating apoM synthesis in vivo. PMID- 15358115 TI - Differential expression and role of two dithiol glutaredoxins Grx1 and Grx2 in Schizosaccharomyces pombe. AB - Glutaredoxins are glutathione-specific thiol oxidoreductases. The regulation and the role of grx1(+) and grx2(+) genes encoding dithiol glutaredoxins were analyzed in Schizosaccharomyces pombe. When tested in the same genetic background including mating type, the grx1 null mutant became sensitive to hydrogen peroxide, whereas grx2 mutant became highly sensitive to paraquat, a superoxide generator. The grx1grx2 double mutant showed additive phenotype of each single mutant. The grx1(+) gene expression was induced by various stresses such as oxidants, salts, and heat, and increased in the stationary phase, whereas grx2(+) stayed constitutive. The induction was mediated via Spc1 MAP kinase path involving both Atf1 and Pap1 transcription factors. Sub-cellular fractionation as well as fluorescence microscopy revealed that Grx1 resides mainly in the cytosol, whereas Grx2 is in mitochondria. These results suggest distinct roles for Grx1 and Grx2 in S. pombe in mediating glutathione-dependent redox homeostasis. PMID- 15358116 TI - The secretable form of trimeric TRAIL, a potent inducer of apoptosis. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a type II transmembrane cytokine molecule of the TNF family. Soluble recombinant TRAIL has been shown to induce apoptosis in a wide variety of cancer cells in vitro and to specifically limit tumor growth without damaging normal cells and tissues in vivo. These results suggest a strong potential of TRAIL as an anticancer therapy. Here we report an artificial TRAIL gene that expresses and secretes trimeric TRAIL into the culture supernatant. This novel TRAIL gene is composed of three functional elements, including a secretion signal, a trimerization domain, and an apoptosis-inducing moiety of TRAIL gene sequence. The expression vectors delivering this TRAIL gene produced secretable forms of trimeric TRAIL proteins. These TRAIL proteins showed greater apoptotic activity than the known TRAIL protein that does not contain an additional trimerization domain. Our data suggest that the gene therapy using our artificial TRAIL gene may be used as an anticancer therapy. PMID- 15358117 TI - Serum amyloid A promotes ABCA1-dependent and ABCA1-independent lipid efflux from cells. AB - Serum amyloid A (SAA) is an acute phase protein that associates with HDL. In order to examine the role of SAA in reverse-cholesterol transport, lipid efflux was tested to SAA from HeLa cells before and after transfection with the ABCA1 transporter. ABCA1 expression increased efflux of cholesterol and phospholipid to SAA by 3-fold and 2-fold, respectively. In contrast to apoA-I, SAA also removed lipid without ABCA1; cholesterol efflux from control cells to SAA was 10-fold higher than for apoA-I. Furthermore, SAA effluxed cholesterol from Tangier disease fibroblasts and from cells after inhibition of ABCA1 by fixation with paraformaldehyde. In summary, SAA can act as a lipid acceptor for ABCA1, but unlike apoA-I, it can also efflux lipid without ABCA1, by most likely a detergent like extraction process. These results suggest that SAA may play a unique role as an auxiliary lipid acceptor in the removal of lipid from sites of inflammation. PMID- 15358118 TI - Regulation of adipocyte differentiation and insulin action with rapamycin. AB - Here, we demonstrated that inhibition of mTOR with rapamycin has negative effects on adipocyte differentiation and insulin signaling. Rapamycin significantly reduced expression of most adipocyte marker genes including PPARgamma, adipsin, aP2, ADD1/SREBP1c, and FAS, and decreased intracellular lipid accumulation in 3T3 L1 and 3T3-F442A cells, suggesting that rapamycin would affect both lipogenesis and adipogenesis. Contrary to the previous report that suppressive effect of rapamycin on adipogenesis is limited to the clonal expansion, we revealed that its inhibitory effect persisted throughout the process of adipocyte differentiation. Thus, it is likely that constitutive activation of mTOR might be required for the execution of adipogenic programming. In differentiated 3T3-L1 adipocytes, chronic treatment of rapamycin blunted the phosphorylation of AKT and GSK, which is stimulated by insulin, and reduced insulin-dependent glucose uptake activity. Taken together, these results suggest that rapamycin not only prevents adipocyte differentiation by decrease of adipogenesis and lipogenesis but also downregulates insulin action in adipocytes, implying that mTOR would play important roles in adipogenesis and insulin action. PMID- 15358119 TI - Antibodies to triosephosphate isomerase in patients with neuropsychiatric lupus. AB - Sera from patients with neuropsychiatric lupus (NP lupus) were screened for antibodies to mouse choroid plexus cell line ECPC-4 by Western blotting. A 29-kDa protein band detected in NP lupus sera was identified as triosephosphate isomerase (TPI). Using Western blotting with TPI, TPI was confirmed as the reactive molecule in sera (6 of 14 samples) and in cerebrospinal fluids (1 of 2 samples) of patients with NP lupus. Enzyme-linked immunosorbent assay with TPI showed that the serum anti-TPI antibody index of 89.8 (SD, 70.1) in NP lupus group was significantly higher than in systemic lupus erythematosus without NP manifestations, 34.6 (29.6); scleroderma, 38.2 (39.9); polymyositis/dermatomyositis, 42.1 (51.5); and control, 31.7 (27.4) groups (p<0.02). Sensitivity, specificity, and positive and negative likelihood ratios of serum anti-TPI antibody index for NP lupus were 42.9%, 94.7%, 8.1%, and 0.6%, respectively. These results suggest that anti-TPI antibodies are closely associated with NP lupus. PMID- 15358120 TI - Cyclin D3 interacts with human activating transcription factor 5 and potentiates its transcription activity. AB - The Cyclin D3 protein is a member of the D-type cyclins. Besides serving as cell cycle regulators, D-type cyclins have been reported to be able to interact with several transcription factors and modulate their transcriptional activations. Here we report that human activating transcription factor 5 (hATF5) is a new interacting partner of Cyclin D3. The interaction was confirmed by in vivo coimmunoprecipitation and in vitro binding analysis. Neither interaction between Cyclin D1 and hATF5 nor interaction between Cyclin D2 and hATF5 was observed. Confocal microscopy analysis showed that Cyclin D3 could colocalize with hATF5 in the nuclear region. Cyclin D3 could potentiate hATF5 transcriptional activity independently of its Cdk4 partner. But Cyclin D1 and Cyclin D2 had no effect on hATF5 transcriptional activity. These data provide a new clue to understand the new role of Cyclin D3 as a transcriptional regulator. PMID- 15358122 TI - A novel preadipocyte cell line established from mouse adult mature adipocytes. AB - We have established a novel preadipocyte cell line from mouse adult mature adipocytes. The mature adipocytes were isolated from fat tissues by taking only the floating population of mature fat cells. The isolated mature adipocytes were de-differentiated into fibroblast-like cells. The in vitro studies showed that the cells could re-differentiate into mature adipocytes after over 20 passages. The in vivo transplantation study also demonstrated that the cells had the full potential to differentiate into mature adipocytes, which has not been shown for the 3T3-L1 preadipocyte cell line derived from mouse embryo. We have further analyzed the expression profile of key fat regulatory genes such as the peroxisome proliferator-activated receptorgamma or CCAAT/enhancer-binding protein gene families. We conclude that our cell line could be used as a preferred alternative to 3T3-L1, potentially reflecting the characteristics of mature adipocytes more, since the cell line is actually derived from adult mature adipocytes. PMID- 15358121 TI - Bax-inhibiting peptide derived from mouse and rat Ku70. AB - Bax is a proapoptotic protein that plays a key role in the induction of apoptosis. Ku70 has activities to repair DNA damage in the nucleus and to suppress apoptosis by inhibiting Bax in the cytosol. We previously designed peptides based on the amino acid sequence of Bax-binding domain of human Ku70, and showed that these peptides bind Bax and inhibit cell death in human cell lines. In the present report, we examined the biological activities of other pentapeptides, VPTLK and VPALR, derived from mouse and rat Ku70. Cells in culture accumulated FITC-labeled VPTLK and VPALR, indicating that these peptides are cell permeable (human, mouse, rat, and porcine cells were examined). These peptides bound to Bax and suppressed cell death in various cell types including primary cultured cells. These data suggest that such Bax inhibiting peptides from three mammalian species may be used to protect healthy cells from apoptotic injury under pathological conditions. PMID- 15358123 TI - Identification of a novel isoform predominantly expressed in gastric tissue and a triple-base pair polymorphism of the cathepsin W gene. AB - In order to investigate the prevalence of potential polymorphisms of the cathepsin W gene, the complete cDNA of 50 dyspeptic patients was analyzed. From those 37 (74%) revealed the wildtype sequence, 6 samples (12%) contained independent single base pair changes including 4 silent and 2 with amino acid changes. Furthermore, a triple-base pair polymorphism was found in 7 samples (14%, 4x heterozygous, 3x homozygous) leading to the following changes: F(217)S, H(248)Y, and I(250)T. Furthermore, a novel alternative splice variant concerning intron 10 was identified in 6 samples (12%). Notably, this novel isoform was only found in samples of gastric mucosa lymphocytes, whereas peripheral NK cells expressed cathepsin W wildtype only. Taken together, this study demonstrated for the fist time that a genetic variant and a novel isoform of cathepsin W are present in about 14% and 12%, respectively, within the Caucasian population. PMID- 15358124 TI - Ectopically expressed gamma-aminobutyric acid receptor B is functionally down regulated in isolated lipid raft-enriched membranes. AB - Lipid raft domains have attracted much recent attention as platforms for plasma membrane signalling complexes. In particular, evidence is emerging that shows them to be key regulators of G protein coupled receptor function. The G protein coupled gamma-aminobutyric acid receptor B (GABA(B) receptor) co-isolates with lipid raft domains from rat brain cerebellum. In the present study, we show that the GABA(B1a,2) receptor was also present in lipid raft domains when expressed ectopically in a Chinese hamster ovary cell line. Lipid raft-associated receptor was functionally active, displaying a concentration-dependent increase in GTPgammaS binding in response to the receptor agonist GABA. Compared with whole cell membranes, lipid raft-associated receptor displayed an increased EC(50) and a reduced magnitude of response to GABA. We conclude that lipid raft association is an intrinsic property of the GABA(B1a,2) receptor and is not cell-type specific. In addition, localisation to lipid raft domains may provide a mechanism to inhibit receptor function. PMID- 15358125 TI - Microbial expression of proteins containing long repetitive Arg-Gly-Asp cell adhesive motifs created by overlap elongation PCR. AB - We developed a novel method for creating repetitive DNA libraries using overlap elongation PCR, and prepared a DNA library encoding repetitive Arg-Gly-Asp (RGD) cell adhesive motifs. We obtained various length DNAs encoding repetitive RGD from a short monomer DNA (18 bp) after a thermal cyclic reaction without a DNA template for amplification, and isolated DNAs encoding 2, 21, and 43 repeats of the RGD motif. We cloned these DNAs into a protein expression vector and overexpressed them as thioredoxin fusion proteins: RGD2, RGD21, and RGD43, respectively. The solubility of RGD43 in water was low and it formed a fibrous precipitate in water. Scanning electron microscopy revealed that RGD43 formed a branched 3D-network structure in the solid state. To evaluate the function of the cell adhesive motifs in RGD43, mouse fibroblast cells were cultivated on the RGD43 scaffold. The fibroblast cells adhered to the RGD43 scaffold and extended long filopodia. PMID- 15358126 TI - Highly infectious SARS-CoV pseudotyped virus reveals the cell tropism and its correlation with receptor expression. AB - Studies of SARS coronavirus (SARS-CoV)-the causative agent of severe acute respiratory syndrome (SARS)-have been hampered by its high transmission rate and the pathogenicity of this virus. To permit analysis of the host range and entry mechanism of SARS-CoV, we incorporated the humanized SARS-CoV spike (S) glycoprotein into HIV particles to generate a highly infectious SARS-CoV pseudotyped virus. The infection on Vero E6-a permissive cell line to SARS-CoV could be neutralized by sera from convalescent SARS patients, and the entry was a pH-dependent process. With these highly infectious SARS-CoV pseudotypes, several cell lines derived from various tissues were revealed as susceptible to SARS-CoV, which were highly corresponding to the expression pattern of virus's receptor angiotensin-converting enzyme 2 (ACE2). In addition, we also demonstrated angiotensin 1 converting enzyme (ACE)-the homologue of ACE2 could not function as a receptor for SARS-CoV. PMID- 15358127 TI - Serum and glucocorticoid inducible kinases functionally regulate ClC-2 channels. AB - ClC-2 participates in the regulation of neuronal excitability, chloride secretion, and cell volume. The ClC-2 sequence contains a consensus site (Ser82) for phosphorylation by the serum and glucocorticoid inducible kinase isoforms SGK1-3. Thus, the present study explored whether ClC-2 is regulated by those kinases. ClC-2 expression in Xenopus oocytes induced inwardly rectifying currents that increased upon coexpression of SGK1-3 and the related kinase PKB. The stimulatory effect was still present upon disruption of the SGK phosphorylation site. SGKs can phosphorylate the ubiquitin ligase Nedd4-2 and prevent Nedd4-2 from binding to its target. Therefore, the role of Nedd4-2 in ClC-2 modulation was investigated. ClC-2 activity decreased upon Nedd4-2 coexpression, an effect reversed by the kinases. According to chemiluminescence ClC-2 membrane abundance was enhanced by SGKs and diminished by Nedd4-2. These observations suggest that SGK1-3 and Nedd4-2 regulate ClC-2 at least in part by modulating ClC-2 abundance at the plasma membrane. PMID- 15358128 TI - Predicting protein structural class by functional domain composition. AB - The functional domain composition is introduced to predict the structural class of a protein or domain according to the following classification: all-alpha, all beta, alpha/beta, alpha+beta, micro (multi-domain), sigma (small protein), and rho (peptide). The advantage by doing so is that both the sequence-order-related features and the function-related features are naturally incorporated in the predictor. As a demonstration, the jackknife cross-validation test was performed on a dataset that consists of proteins and domains with only less than 20% sequence identity to each other in order to get rid of any homologous bias. The overall success rate thus obtained was 98%. In contrast to this, the corresponding rates obtained by the simple geometry approaches based on the amino acid composition were only 36-39%. This indicates that using the functional domain composition to represent the sample of a protein for statistical prediction is very promising, and that the functional type of a domain is closely correlated with its structural class. PMID- 15358129 TI - A transcriptional repressor of osteopontin expression in the 4T1 murine breast cancer cell line. AB - Osteopontin (OPN) is a highly hydrophilic and negatively charged sialoprotein of approximately 298 amino acids which is an important mediator of tumor metastatic behavior. We have previously demonstrated that endotoxin-dependent OPN gene transcription is regulated by a constitutive transcriptional repressor protein, heterogeneous nuclear ribonucleoprotein A/B (hnRNP-A/B). However, in the context of cancer, the role of hnRNP-A/B in the transcriptional regulation of OPN and its metastasis-promoting functions has not been previously studied. We examined hnRNP A/B in the 4T1 murine mammary epithelial tumor cell line, a thioguanine resistant subline which closely mimics stage IV breast cancer in humans. Our data indicate that hnRNP-A/B p37 binds to the OPN promoter, significantly decreases OPN promoter activity and mRNA levels, ablates OPN protein expression, and inhibits OPN dependent in vitro correlates of metastatic behavior, motility, and invasion. These results are unique and may suggest new therapies to re-establish loco regional control of cancers. PMID- 15358130 TI - Concurrent targeted exchange of three bases in mammalian hprt by oligonucleotides. AB - The repair of point mutations in hprt gene by single-stranded oligonucleotides represents a model to test targeted nucleotide exchange. We studied the concurrent nucleotide exchange of two or three nucleotides in the hprt deficient hamster cell line V79-151. The used oligonucleotides resulted in mismatches at two (151, 159) or three (151, 144, and 159) hprt positions. The hprt point mutation at position 151 was repaired in about 2/10(6) cells as shown by hprt sequencing in clones surviving HAT selection. The second nucleotide exchange at hprt position 159 was found in 7% of these HAT selected clones. Using oligonucleotides resulting in three mismatches, 29% of the clones showed nucleotide exchanges at the two hprt positions (151, 144) and about 4% at three positions (151, 144, and 159). These results indicate that single-stranded oligonucleotides can generate two or three nucleotide exchanges in a mammalian chromosomal gene. PMID- 15358131 TI - GDF-3 is an adipogenic cytokine under high fat dietary condition. AB - Growth differentiation factor 3 (GDF-3) is structurally a bone morphogenetic protein/growth differentiation factor subfamily member of the TGF-beta superfamily. GDF-3 exhibits highest level of expression in white fat tissue in mice and is greatly induced by high fat diet if fat metabolic pathway is blocked. To identify its biological function, GDF-3 was overexpressed in mice by adenovirus mediated gene transfer. Mice transduced with GDF-3 displayed profound weight gain when fed with high fat diet. The phenotypes included greatly expanded adipose tissue mass, increased body adiposity, highly hypertrophic adipocytes, hepatic steatosis, and elevated plasma leptin. GDF-3 stimulated peroxisome proliferator activated receptor expression in adipocytes, a master nuclear receptor that controls adipogenesis. However, GDF-3 was not involved in blood glucose homeostasis or insulin resistance, a condition associated with obesity. In contrast, similar phenotypes were not observed in GDF-3 mice fed with normal chow, indicating that GDF-3 is only active under high lipid load. Thus, GDF-3 is a new non-diabetic adipogenic factor tightly coupled with fat metabolism. PMID- 15358132 TI - Posttranscriptional regulation of the karyogamy gene by Kem1p/Xrn1p exoribonuclease and Rok1p RNA helicase of Saccharomyces cerevisiae. AB - The major biochemical activities ascribed to Kem1p/Xrn1p of Saccharomyces cerevisiae are 5'-3' exoribonuclease functioning in RNA turnover and a microtubule-binding protein. Mutational analysis has shown that Kem1p/Xrn1p participates in microtubule-related functions such as nuclear fusion (karyogamy) during mating, chromosome transmission, and spindle pole body duplication. Here, evidence is presented that Kem1p plays a specific role in nuclear fusion by affecting, at the posttranscriptional level, the pheromone induction of the karyogamy-specific transcription factor Kar4p and the expression of Rok1p, a putative RNA helicase. We found that Rok1p itself also affects the pheromone induction of Kar4p and thereby participates in nuclear fusion. Analysis of the active-site mutations, xrn1-D206A or D208A, shows that nuclear fusion as well as the Rok1p synthesis do not require the exoribonuclease activity of Kem1p. Our data provide an important insight into the gene-specific regulatory function mediated by the general RNA-modulating enzymes. PMID- 15358133 TI - 2-Dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole: a novel powerful and selective inhibitor of protein kinase CK2. AB - Protein kinase CK2 is a highly pleiotropic enzyme whose high constitutive activity is suspected to be instrumental to the enhancement of the tumour phenotype and to the propagation of infectious diseases. Here we describe a novel compound, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT), which is superior to the commonly used specific CK2 inhibitor 4,5,6,7 tetrabromobenzotriazole (TBB) in several respects. DMAT displays the lowest K(i) value ever reported for a CK2 inhibitor (40 nM); it is cell permeable and its efficacy on cultured cells, both in terms of endogenous CK2 inhibition and induction of apoptosis, is several fold higher than that of TBB. The selectivity of DMAT assayed on a panel of >30 protein kinases is comparable to that of TBB, with the additional advantage of being ineffective on protein kinase CK1 up to 200 microM. These properties make DMAT the first choice CK2 inhibitor for in vivo studies available to date. PMID- 15358134 TI - Neuroglycan C, a novel member of the neuregulin family. AB - Neuroglycan C (NGC) is a transmembrane chondroitin sulfate proteoglycan expressed predominantly in the brain that possesses an EGF-like extracellular domain. The goal of the present study was to determine whether NGC may activate ErbB tyrosine kinases. A recombinant human NGC extracellular domain induced tyrosine phosphorylation of ErbB2 and ErbB3 as well as cell growth of the human breast tumor cell lines, T47D and MDA-MB-453. In vitro pull-down assay revealed that NGC could directly bind to a recombinant ErbB3-immunoglobulin Fc fusion protein (ErbB3-Fc) but not to ErbB1-Fc, ErbB2-Fc or ErbB4-Fc. A newly established anti ErbB3 neutralizing monoclonal antibody (#5C3) almost completely blocked NGC induced ErbB activation in MDA-MB-453 cells. Taken together, these data indicate that NGC is an active growth factor and a direct ligand for ErbB3 and that NGC transactivates ErbB2. Thus, NGC should be classified as the sixth member (neuregulin-6) of the neuregulin family. PMID- 15358135 TI - Mac-1(low) early myeloid cells in the bone marrow-derived SP fraction migrate into injured skeletal muscle and participate in muscle regeneration. AB - Recent studies have shown that bone marrow (BM) cells, including the BM side population (BM-SP) cells that enrich hematopoietic stem cells (HSCs), are incorporated into skeletal muscle during regeneration, but it is not clear how and what kinds of BM cells contribute to muscle fiber regeneration. We found that a large number of SP cells migrated from BM to muscles following injury in BM transplanted mice. These BM-derived SP cells in regenerating muscles expressed different surface markers from those of HSCs and could not reconstitute the mouse blood system. BM-derived SP/Mac-1(low) cells increased in number in regenerating muscles following injury. Importantly, our co-culture studies with activated satellite cells revealed that this fraction carried significant potential for myogenic differentiation. By contrast, mature inflammatory (Mac-1(high)) cells showed negligible myogenic activities. Further, these BM-derived SP/Mac-1(low) cells gave rise to mononucleate myocytes, indicating that their myogenesis was not caused by stochastic fusion with host myogenic cells, although they required cell-to-cell contact with myogenic cells for muscle differentiation. Taken together, our data suggest that neither HSCs nor mature inflammatory cells, but Mac-1(low) early myeloid cells in the BM-derived SP fraction, play an important role in regenerating skeletal muscles. PMID- 15358136 TI - Evidence that Plasmodium falciparum diacylglycerol acyltransferase is essential for intraerythrocytic proliferation. AB - In triacylglycerol (TAG)-accumulating organisms, the physiological roles of diacylglycerol acyltransferase (DGAT), a principal enzyme in the major biosynthetic pathway for TAG, appear to be diverse. Apicomplexan parasite, Plasmodium falciparum, shows unique features in TAG metabolism and trafficking during intraerythrocytic development, and unlike most eukaryotes, only one open reading frame (ORF) encoding a candidate DGAT could be found in its genome. However, whether this candidate ORF encodes P. falciparum DGAT and its physiological relevance have not been assessed. Here, we demonstrate that the ORF is transcribed as a approximately 3.6 kb single mRNA throughout intraerythrocytic development, markedly elevated at trophozoite, schizont, and segmented schizont, and indeed encodes a protein exhibiting DGAT activity. Further, we provide evidence that the parasite in which the ORF was disrupted via double crossover recombination cannot be enriched, implying a fundamental role of PfDGAT in intraerythrocytic proliferation. PMID- 15358137 TI - Quantum algorithm for programmed cell death of Caenorhabditis elegans. AB - During the development of Caenorhabditis elegans, through cell divisions, a total of exactly 1090 cells are generated, 131 of which undergo programmed cell death (PCD) to result in an adult organism comprising 959 cells. Of those 131, exactly 113 undergo PCD during embryogenesis, subdivided across the cell lineages in the following fashion: 98 for AB lineage; 14 for MS lineage; and 1 for C lineage. Is there a law underlying these numbers, and if there is, what could it be? Here we wish to show that the count of the cells undergoing PCD complies with the cipher laws related to the algorithms of Shor and of Grover. PMID- 15358138 TI - ZAP is a CRM1-dependent nucleocytoplasmic shuttling protein. AB - The zinc finger antiviral protein (ZAP) is a recently isolated host antiviral factor. It specifically inhibits the replication of Moloney murine leukemia virus (MMLV) and Sindbis virus (SIN) by preventing the accumulation of viral RNA in the cytoplasm. In this report, we demonstrate that ZAP is predominantly localized in the cytoplasm at steady state but shuttles between the nucleus and the cytoplasm in a CRM1-dependent manner. Two nuclear localization sequences (NLS) and one nuclear export sequence (NES) were identified. One NLS was mapped to amino acids 68-RARVCRRK-75 and the other mapped to a region including amino acids K405 and K406. The NES was mapped to amino acids 284-LEDVSVDV-291. These findings help to understand why ZAP specifically prevents the accumulation of viral RNA in the cytoplasm. These findings also suggest possible functions of ZAP in the nucleus. PMID- 15358139 TI - Co-inhibition of epidermal growth factor receptor and type 1 insulin-like growth factor receptor synergistically sensitizes human malignant glioma cells to CD95L induced apoptosis. AB - Inhibition of epidermal growth factor receptor (EGFR) signaling sensitizes human malignant glioma cells to death ligand-induced apoptosis. However, tumor cells may compensate the loss of EGFR signaling by activation of the type 1 insulin like growth factor receptor (IGF-1R). We here report that antagonism of the IGF 1R with the small-molecule inhibitor AG1024 in combination with inhibitors of the EGFR synergistically sensitizes human malignant glioma cells to CD95L-induced apoptosis. This cell death is p53-independent, but requires caspase 8 activity. The levels of the receptor, CD95, are not altered by the inhibitors alone or in combination. Analysis of the downstream signaling pathways reveals synergistic inhibition of ribosomal protein S6 phosphorylation by inhibitor co-treatment, suggesting an involvement of the mammalian target of rapamycin pathway. These findings suggest that adding inhibitors of IGF-1R may be a strategy to overcome escape from the anti-apoptotic effects of EGFR inhibition in malignant gliomas. PMID- 15358140 TI - Proprotein convertases regulate insulin-like growth factor 1-induced membrane type 1 matrix metalloproteinase in VSMCs via endoproteolytic activation of the insulin-like growth factor-1 receptor. AB - The IGF-1 receptor (IGF-1R) and MT1-MMP are synthesized as larger precursor proproteins, which require endoproteolytic activation by the proprotein convertases (PCs) furin/PC5 to gain full biological activity. The aim of this study was to investigate the contribution of PCs to IGF-1R and/or MT1-MMP activation in vascular smooth muscle cells (VSMCs) as well as VSMC proliferation/migration, which are key elements in vascular remodeling. Furin and PC5 mRNAs and proteins were found in VSMCs. Inhibition of furin-like PCs with the specific pharmacological inhibitor dec-CMK inhibited IGF-1R endoproteolytic activation. Inhibition of IGF-1R maturation abrogated IGF-induced IGF-1R autophosphorylation, PI3-kinase and MAPK induction, as well as VSMC proliferation (p<0.05 vs. controls), whereas it had no effect of PDGF-stimulated signaling pathways or cell growth. Both, IGF-1 and PDGF-BB, induced MT1-MMP expression, but only IGF-1-mediated MT1-MMP induction was inhibited by dec-CMK. Induction of MMP 2 by IGF-1 was inhibited by the PI3-kinase inhibitor wortmannin, but not by the MEK-inhibitor PD98059. Dec-CMK inhibited VSMC chemotaxis comparable to the effects of the MMP-inhibitor GM6001 (both p<0.05 vs. controls), supporting that MMPs are involved. In conclusion, this study demonstrates that targeting furin like PCs and thus inhibiting IGF-1R activation is a novel target to inhibit IGF-1 mediated signaling and cell functions, such as IGF-1-induced MT1-MMP/MMP-2 in VSMCs. PMID- 15358141 TI - Molecular mechanisms of echinocystic acid-induced apoptosis in HepG2 cells. AB - Echinocystic acid (EA), a natural triterpone enriched in various herbs, has been showed to have cytotoxic activity in some cancer cells, and is used for medicinal purpose in many Asian countries. In the present study, we found that EA could induce apoptosis in human HepG2 cells, as characterized by DNA fragmentation, activation of caspase-3, -8, and -9, and PARP cleavage. The efficacious induction of apoptosis was observed at 45 microM for 24 h. Molecular data showed that EA induced the truncation of Bid protein and reduction of Bcl-2 protein. EA also caused the loss of mitochondrial membrane potential (DeltaPsi(m)) and cytochrome c release from mitochondria to cytosol. Moreover, EA could activate c-Jun NH(2) terminal kinase (JNK) and p38 kinase, and JNK-specific inhibitor SP600125 and p38 kinase-specific inhibitor SB200235 could block serial molecular events of EA induced apoptosis such as Bid truncation, Bcl-2 reduction, cytochrome c release, caspase activation, and DNA fragmentation in HepG2 cells. These findings indicate that JNK- and p38 kinase-mediated mitochondrial pathways might be involved in EA induced apoptosis and enhance our understanding of the anticancer function of EA in herbal medicine. PMID- 15358142 TI - Post genomic analysis of permeases from the amino acid/auxin family in protozoan parasites. AB - The "amino acid/auxin permeases" is probably the most represented family of transporters in the Trypanosoma cruzi genome. Using a high-throughput searching routine and preliminary data from the T. cruzi genome project, more than 15,000 sequences were iteratively assembled into contigs, and 60 open reading frames corresponding to different putative amino acid transporters, clustered in 12 groups, were detected and characterized in silico. T. cruzi genomic organization of such sequences showed that these putative amino acid transporter genes are in an unusually large number and arranged in repeat clusters comprising about 0.2% of the genome. These data suggest that the family has evolved following tandem duplication events and constitutes a novel family of variable proteins in protozoan organisms. The mRNA expression of the predicted genes was demonstrated in infective and non-infective parasite forms. Orthologous sequences were also identified in other unicellular parasites such as Leishmania spp., Plasmodium spp., and Trypanosoma brucei. PMID- 15358143 TI - Nucleocapsid protein of SARS coronavirus tightly binds to human cyclophilin A. AB - Severe acute respiratory syndrome coronavirus (SARS-CoV) is responsible for SARS infection. Nucleocapsid protein (NP) of SARS-CoV (SARS_NP) functions in enveloping the entire genomic RNA and interacts with viron structural proteins, thus playing important roles in the process of virus particle assembly and release. Protein-protein interaction analysis using bioinformatics tools indicated that SARS_NP may bind to human cyclophilin A (hCypA), and surface plasmon resonance (SPR) technology revealed this binding with the equilibrium dissociation constant ranging from 6 to 160nM. The probable binding sites of these two proteins were detected by modeling the three-dimensional structure of the SARS_NP-hCypA complex, from which the important interaction residue pairs between the proteins were deduced. Mutagenesis experiments were carried out for validating the binding model, whose correctness was assessed by the observed effects on the binding affinities between the proteins. The reliability of the binding sites derived by the molecular modeling was confirmed by the fact that the computationally predicted values of the relative free energies of the binding for SARS_NP (or hCypA) mutants to the wild-type hCypA (or SARS_NP) are in good agreement with the data determined by SPR. Such presently observed SARS_NP-hCypA interaction model might provide a new hint for facilitating the understanding of another possible SARS-CoV infection pathway against human cell. PMID- 15358144 TI - HIV-1 and MLV Gag proteins are sufficient to recruit APOBEC3G into virus-like particles. AB - The cytidine deaminase hAPOBEC3G is an antiviral human factor that counteracts the replication of HIV-1 in absence of the Vif protein. hAPOBEC3G is packaged into virus particles and lethally hypermutates HIV-1. In this work, we examine the mechanisms governing hAPOBEC3G packaging. By GST pull-down and co immunoprecipitation assays, we show that hAPOBEC3G binds to HIV-1 Pr55 Gag and its NC domain and to the RT and IN domains contained in Pr160 Gag-Pol. We demonstrate that the expression of HIV-1 Gag is sufficient to induce the packaging of hAPOBEC3G into Gag particles. Gag-Pol polypeptides containing RT and IN domains, as well as HIV-1 genomic RNA, seem not to be necessary for hAPOBEC3G packaging. Lastly, we show that hAPOBEC3G and its murine ortholog are packaged into HIV-1 and MLV Gag particles. We conclude that the Gag polypeptides from distant retroviruses have conserved domains allowing the packaging of the host antiviral factor APOBEC3G. PMID- 15358145 TI - Co-chaperone CHIP associates with mutant Cu/Zn-superoxide dismutase proteins linked to familial amyotrophic lateral sclerosis and promotes their degradation by proteasomes. AB - Although the ubiquitin-proteasome system and the molecular chaperones are implicated to play an important role in pathogenesis of familial amyotrophic lateral sclerosis (FALS) caused by mutations in Cu/Zn-superoxide dismutase (SOD1), the mechanism underlying the causes of this fatal disease is still poorly understood. Here we found that co-chaperone CHIP (carboxyl terminus of Hsc70 interacting protein), together with molecular chaperones Hsc70/Hsp70 and Hsp90, associates with FALS-linked mutant SOD1 proteins in cultured human cells. S5a subunit of 26S proteasomes, which recognizes polyubiquitylated proteins, also interacts with mutant SOD1 proteins. Over-expression of CHIP leads to the reduction in cellular levels of mutant SOD1 as well as the suppression of cytotoxicity induced by mutant SOD1. Unusually, rather than increasing the level of poly-ubiquitylated SOD1, over-expressed CHIP alters the ubiquitylation pattern of mutant SOD1 proteins. Both down-regulation and ubiquitylation of mutant SOD1 are greatly reduced by a mutant CHIP protein lacking U-box domain. Taken together, these results suggest that co-chaperone CHIP, possibly with another E3 ligase(s), modulates the ubiquitylation of mutant SOD1 and renders them more susceptible for proteasomal degradation. PMID- 15358146 TI - Characterization of the aspartate kinase from Saccharomyces cerevisiae and of its interaction with threonine. AB - Aspartate kinase (AK) from Saccharomyces cerevisiae has been characterized to elucidate its quaternary structure and the effect of the allosteric inhibitor threonine on the enzyme conformation. The homogeneously purified enzyme was inhibited by threonine (K(i) 1.4 mM) and was found to bind this compound (K(d) 0.97 mM) in a hyperbolic manner. Gel filtration and native gel electrophoresis indicated that yeast AK is a homohexamer of 346 kDa composed by 58 kDa subunits. Threonine caused a decrease in the apparent molecular mass of AK as evidenced by size-exclusion chromatography (from 345 to 280 kDa) and blue native gel electrophoresis (from 346 to 297 kDa); no other molecular species were detected. This shift in the hydrodynamic size was threonine-specific and was reversed by rechromatography in the absence of threonine. No change in the apparent molecular mass was induced by threonine in an AK mutant insensitive to inhibition by this amino acid, which was observed to be unable to bind threonine. These results indicate that the allosteric transition elicited by binding of threonine to yeast AK involves a large conformational change of the protein that isomerizes from a relaxed active conformation to a more compact inactive one of smaller molecular dimensions. PMID- 15358147 TI - Inhibition of lipopolysaccharide-mediated human monocyte activation, in vitro, by alpha1-antitrypsin. AB - alpha1-Antitrypsin (AAT) is a major circulating and tissue inhibitor of serine proteinases. As such AAT is thought to play an important role in limiting host tissue injury at sites of inflammation. There is now increasing evidence, however, that AAT may exhibit biological activity independent of its protease inhibitor function. In this study we compared the effects of native (inhibitory) and modified (non-inhibitory), e.g., polymerised and oxidised forms of AAT on LPS induced human monocyte activation, in vitro. We found that native AAT inhibited LPS-stimulated synthesis and release of TNFalpha and IL-1beta mRNA and protein, respectively, but enhanced the release of the anti-inflammatory cytokine, IL-10. Similarly, polymerised and oxidised forms of AAT inhibited LPS-stimulated IL 1beta and TNFalpha. The effects of AATs were observed whether added prior to or following removal of LPS, suggesting that sequestration of agonist was unlikely to explain their biological effects. Furthermore, studies with neutralising antibodies indicated that generation of IL-10 was unlikely to be the mechanism responsible for the inhibitory effects of AATs. Thus, our data demonstrate for the first time that AAT exhibits anti-inflammatory activity in vitro that is unrelated to inhibition of serine proteases. PMID- 15358149 TI - Protein-protein interactions of KChIP proteins and Kv4.2. AB - To prove heteromeric assembly of KChIP proteins, the present study is carried out. The results of chemical crosslinking and pull down assay revealed that KChIP1, KChIP2.1, and KChIP2.2 could form homo- as well as hetero-oligomer, and this oligomerization exhibited a Ca(2+)-dependent manner. Moreover, homomeric and heteromeric assembly of KChIPs did not perturb their interaction with Kv4.2 K(+) channel, indicating that the region associated with oligomerization of KChIPs was distinct from that for binding with Kv4.2. Together with previous findings that the net effects of KChIP proteins on the molecular properties and trafficking of Kv channel were different, these observations open a fascinating possibility that the electrophysiological properties of Kv channel may be differently regulated by homomeric and heteromeric assembly of KChIPs. PMID- 15358148 TI - Cross-inhibition between furin and lethal factor inhibitors. AB - Bacillus anthracis synthesizes two toxins composed of the three proteins: protective antigen (PA), lethal factor (LF), and edema factor (EF). The cleavage of PA on the cell surface by the convertase furin leads to the translocation of LF and EF into the cytosol. We have investigated the cross-inhibitory activities of the furin inhibitors hexa-d-arginine amide (D6R) and nona-d-arginine amide (D9R), which block the proteolytic activation of PA; and of the LF inhibitor In-2 LF, a peptide hydroxamate. D6R and D9R inhibit LF with IC(50s) of 300 and 10microM, respectively; conversely, In-2-LF also inhibits furin (IC(50) 2microM). In-2-LF was efficiently cleaved by furin with the concomitant loss of inhibitory activity on both LF and furin. Incubation of In-2-LF with LF however generated a product that retained partial inhibitory activity against LF. Combined treatment of cells with D6R and In-2-LF enhanced protection against anthrax lethal toxin, indicating that combined administration of inhibitors could represent an effective therapeutic approach. PMID- 15358150 TI - Suppression of the cell proliferation in stomach cancer cells by the ZNRD1 gene. AB - Zinc ribbon domain-containing 1 (ZNRD1), a transcription-associated gene, was recently found to be downregulated in human gastric cancer tissues as compared to the matched adjacent nonneoplastic tissues. In this study, we constructed the siRNA eukaryotic expression vectors of ZNRD1 and transfected them into normal gastric epithelial cells (GES-1). We also introduced the ZNRD1 gene into gastric cancer cells that do (SGC7901) and do not (AGS) express ZNRD1 endogenously. GES-1 cells stably transfected with the ZNRD1-RNAi were found to exhibit significantly quicker proliferation than empty vector transfectants. AGS cells stably transfected with the ZNRD1 cDNA exhibited significantly decreased growth rate as compared to control vector transfectants, whereas SGC7901 cells did not. Furthermore, ZNRD1 suppresses growth of AGS cells in soft agar and tumor formation in athymic nude mice. This study clearly demonstrates that ZNRD1 may play an important role in the control of human gastric cancer development by regulating cell proliferation. These results provide new insights into the function of ZNRD1 and further validate ZNRD1 as a potential therapeutic target in gastric cancer. PMID- 15358151 TI - Distending stress of the cytoskeleton is a key determinant of cell rheological behavior. AB - One fundamental question in cell biology is what determines rheological properties of living cells. If the cytoskeletal distending stress is a key determinant of cell rheology, then modulating this stress by cell stretching should have a major effect on cell rheological properties. If not, then other mechanisms must play a major role. We developed a stretchable cell culture device that could rapidly stretch cells and thus generate passive mechanical stress within the cytoskeleton. This device was placed inside a magnetic cytometry system to measure the effect of stretching on rheological properties of cultured human airway smooth muscle cells. A gradual increase in cell distension caused a systematic increase in cell dynamic stiffness in a manner which was consistent with earlier observations where the active component of the distending stress was modulated pharmacologically. These findings provide strong evidence that the cytoskeletal distending stress is a key determinant of cell rheological properties. PMID- 15358152 TI - Actin is closely associated with RNA polymerase II and involved in activation of gene transcription. AB - Biochemical and morphological studies have demonstrated the presence of actin in the nucleus of different eukaryotic cells, whereas its role remains unclear. In this work, we studied the interaction and the functional relationship between nuclear actin and RNA polymerase II (RNAP II). The immunofluorescence study demonstrated a clear co-localization of nuclear actin with RNAP II in HeLa cells. Meanwhile, actin can be immunoprecipitated by anti-RNAP II antibody, indicating that they could interact with each other. Treatment of cells with alpha-amanitin induced the formation of actin bundle network in the nucleoplasm. Blocking of the formation of filamentous actin (F-actin) by cytochalasin B modified the distribution of actin. Although the actin content remained unchanged in resting and concanavalinA stimulated mouse lymphocytes, the actin content in the nuclei showed a progressive increase after stimulation. Furthermore, the antibody against actin blocked RNA synthesis in a eukaryotic in vitro transcription system. These observations implicate that nuclear actin interacts with RNAP II and may have function on the RNAP II-mediated transcription. PMID- 15358153 TI - Antibiotic activity and synergistic effect of antimicrobial peptide against pathogens from a patient with gallstones. AB - HP (2-20) is a peptide derived from the N-terminus of Helicobacter pylori ribosomal protein L1 that has been shown to have antimicrobial activity against various species of bacteria. When we tested the effects of HP (2-20), we found that this peptide displayed strong activity against pathogens from a patient with gallstones, but it did not have hemolytic activity against human erythrocytes. We also found that HP (2-20) had potent activity against cefazolin sodium-resistant bacterial cell lines, and that HP (2-20) and cefazolin sodium had synergistic effects against cell lines resistant to the latter. To investigate the mechanism of action of HP (2-20), we performed fluorescence activated flow cytometry using pathogens from the patient with gallstones. As determined by propidium iodide (PI) staining, pathogenic bacteria treated with HP (2-20) showed higher fluorescence intensity than untreated cells, similar to melittin-treated cells, and that HP (2-20) acted in an energy- and salt-dependent manner. Scanning electron microscopy showed that HP (2-20) caused significant morphological alterations in the cell surface of pathogens from the patient with gallstones. By determining their 16S rDNA sequences, we found that both the pathogens from the patient with gallstones and the cefazolin sodium-resistant cell lines showed 100% homology with sequences from Pseudomonas aeruginosa. Taken together, these results suggest that HP (2-20) has antibiotic activity and that it may be used as a lead drug for the treatment of acquired pathogens from patients with gallstones and antibiotic-resistant cell lines. PMID- 15358154 TI - Over-expression of SR-cyclophilin, an interaction partner of nuclear pinin, releases SR family splicing factors from nuclear speckles. AB - Pre-mRNA splicing takes place within a dynamic ribonucleoprotein particle called the spliceosome and occurs in an ordered pathway. Although it is known that spliceosome consists of five small nuclear RNAs and at least 50 proteins, little is known about how the interaction among the proteins changes during splicing. Here we identify that SR-cyp, a Moca family of nuclear cyclophilin, interacts and colocalizes with nuclear pinin (pnn), a SR-related protein involving in pre-mRNA splicing. Nuclear pnn interacts with SR-cyp via its C-terminal RS domain. Upon SR cyp over-expression, however, the subnuclear distribution of nuclear pnn is altered, resulting in its redistribution from nuclear speckles to a diffuse nucleoplasmic form. The diffuse subnuclear distribution of nuclear pnn is not due to epitope masking, accelerated protein turnover or post-translational modification. Furthermore, we find that SR-cyp regulates the subnuclear distribution of other SR family proteins, including SC35 and SRm300, in a similar manner as it does on nuclear pnn. This result is significant because it suggests that SR-cyp plays a general role in modulating the distribution pattern of SR like and SR proteins, similar to that of Clk (cdc2-like kinase)/STY on SR family splicing factors. SR-cyp might direct its effect via either alteration of protein folding/conformation or of protein-protein interaction and thus may add another control level of regulation of SR family proteins and modification of their functions. PMID- 15358155 TI - VEGF selectively induces Down syndrome critical region 1 gene expression in endothelial cells: a mechanism for feedback regulation of angiogenesis? AB - The Down syndrome critical region 1 (DSCR1) gene (also known as MCIP1, Adapt78) encodes a regulatory protein that binds to calcineurin catalytic A subunit and acts as a regulator of the calcineurin-mediated signaling pathway. We show in this study that DSCR1 is greatly induced in endothelial cells in response to VEGF, TNF-alpha, and A23187 treatment, and that this up-regulation is inhibited by inhibitors of the calcineurin-NFAT (nuclear factor of activated T cells) signaling pathway as well as by PKC inhibition and a Ca(2+) chelator. We hypothesized that the up-regulation of DSCR1 gene expression in endothelial cells could act as an endogenous feedback inhibitor for angiogenesis by regulating the calcineurin-NFAT signaling pathway. Our transient transfection analyses confirm that the overexpression of DSCR1 abrogates the up-regulation of reporter gene expression driven by both the cyclooxygenase 2 and DSCR1 promoters in response to stimulators. Our results indicate that DSCR1 up-regulation may represent a potential molecular mechanism underlying the regulation of angiogenic genes activated by the calcineurin-NFAT signaling pathway in endothelial cells. PMID- 15358156 TI - A pathway involving protein kinase Cdelta up-regulates cytosolic phospholipase A(2)alpha in airway epithelium. AB - Cytosolic phospholipase A(2)alpha (cPLA(2)alpha) catalyzes the hydrolysis of glycerophospholipids at the sn-2 position to liberate fatty acids. Although cPLA(2)alpha has been implicated in various cellular processes, the detailed mechanism of its expression remains to be elucidated. Here we report that phorbol 12-myristate 13-acetate (PMA) up-regulates cPLA(2)alpha in A549 airway epithelium cells, and that this effect is sensitive to rottlerin, a potent inhibitor of protein kinase Cdelta (PKCdelta). Consistent with this observation, a dominant negative mutant of PKCdelta reduced cPLA(2)alpha induction in response to PMA. Up regulation of cPLA(2)alpha by PMA was also inhibited by PDTC, an inhibitor of nuclear factor-kappa B (NF-kappaB), and degradation of IkappaB and subsequent activation of NF-kappaB occurred in response to PMA treatment. These findings indicate that PMA induces expression of cPLA(2)alpha at the transcriptional level via an NF-kappaB-dependent mechanism. In addition, activation of the NF-kappaB promoter by PMA was diminished by pretreatment with DPI, a flavoenzyme inhibitor as well as by rottlerin, suggesting a role for reactive oxygen species (ROS) as well as PKCdelta. Consistent with this, PMA stimulated the production of ROS and this was blocked by inhibiting PKCdelta. Our results suggest that PKCdelta and ROS lie upstream of NF-kappaB, and we conclude that a PKCdelta-ROS-NF-kappaB cascade plays a pivotal role in cPLA(2)alpha induction by PMA. PMID- 15358157 TI - Geldanamycin induces Hsp70 and prevents alpha-synuclein aggregation and toxicity in vitro. AB - Geldanamycin (GA) is a naturally occurring benzoquinone ansamycin that induces heat shock protein 70 (Hsp70). GA has been shown to reduce alpha-synuclein induced neurotoxicity in a fly model of Parkinson's disease. We have previously shown that heat shock proteins can prevent alpha-synuclein aggregation and protect against alpha-synuclein induced toxicity in human H4 neuroglioma cells. Here, we hypothesize that GA treatment will reduce alpha-synuclein aggregation and prevent alpha-synuclein induced toxicity and we show that GA can induce Hsp70 in a time- and concentration-dependent manner in H4 cells. Pretreatment with 200nM GA 24h prior to transfection prevented alpha-synuclein aggregation and protected against toxicity. Treatment of cells with pre-existing inclusions with GA did not result in a reduction in the number of cells containing inclusions, suggesting that upregulation of Hsp70 is not sufficient to remove established inclusions. Similarly, Western blot analysis demonstrated that GA treatment could dramatically reduce both total alpha-synuclein and high molecular weight alpha synuclein aggregates. Taken together, these data suggest that GA is effective in preventing alpha-synuclein aggregation and may represent a pharmacological intervention to therapeutically increase expression of molecular chaperone proteins to treat neurodegenerative diseases where aggregation is central to the pathogenesis. PMID- 15358158 TI - Pitavastatin increases ABCA1-mediated lipid efflux from Fu5AH rat hepatoma cells. AB - ATP binding cassette A1 (ABCA1) is responsible in vivo for the formation of HDL by promoting the lipidation of apoprotein A-I (apoA-I) via cholesterol and phospholipid efflux from the liver. Treatment of patients with statins produces an increase in HDL plasma level, but the underlying mechanism is not completely understood. In this work we investigated the ability of pitavastatin to modulate ABCA1-mediated efflux from Fu5AH rat hepatoma cells, that here we demonstrate to express functional ABCA1 upon treatment with 22OH/cRA. In both basal and ABCA1 expressing cells pitavastatin 0.1-50microM induced a dose-dependent increase in cholesterol efflux to apoA-I; this effect was reversed by mevalonate or geranyl geraniol. A stimulatory effect was also observed on phospholipid efflux. Similar results were obtained with compactin, suggesting a class-related effect of statins. These results indicate a potential mechanism for the improvement in HDL plasma profile observed in patients treated with statins. PMID- 15358159 TI - MEK inhibition and phosphorylation of serine 4 on B23 are two coincident events in mitosis. AB - Previous studies have shown that activation of the Raf/MEK/ERK pathway is necessary for G2/M transition. However, as for the activation state of MEK in mitosis the conclusion is not consistent. Here we show that MEK is inhibited in mitosis. In addition, we identify a multifunctional protein named B23 that strongly cross-reacts with a phospho-MEK antibody in mitotic cells. Sequence homology between the N-terminus surrounding Ser 4 of B23 and the Raf phosphorylation site on MEK suggests a mechanism for cross-reaction of the antibody. Thus, mutation of Ser 4 to alanine abolishes cross-reactivity between B23 and the phospho-MEK antibody. Our findings may explain the discrepancy of results obtained with the use of phospho-MEK antibody regarding the activation state of MEK in mitosis. PMID- 15358160 TI - Cell adhesion to ephrinb2 is induced by EphB4 independently of its kinase activity. AB - Cell to cell interaction in bone marrow is crucial for differentiation of hematopoietic cells. We have shown that EphB4 receptor is expressed in erythroid progenitor and its activation accelerates erythroid differentiation. To elucidate the role of EphB4 activation in erythropoiesis, we analyzed effects of EphB4 on cell adhesive pathways. Cell adhesion with the extension of filopodial pseudopod was observed by EphB4 activation. EphB4 activation also enhanced an effect of fibronectin-mediated adhesive pathway along with formation of the c-Cbl/CrkL complex. The tyrosine kinase activity of EphB4 was dispensable for those phenomena. These results suggest that activation of EphB4 participates in adhesive but not repulsive signals independently of its tyrosine kinase activity in hematopoietic cells. PMID- 15358161 TI - Nitric oxide and TGF-beta1 inhibit HNF-4alpha function in HEPG2 cells. AB - This study analyzes if the profibrogenic factors nitric oxide and transforming growth factor-beta1 (TGF-beta1) affect hepatocyte nuclear factor-4alpha (HNF 4alpha) function. For this purpose, HepG2 cells were treated with TGF-beta1 or with a nitric oxide donor to determine mRNA levels of coagulation factor VII and HNF-4alpha. Treatment effect on factor VII gene promoter was assessed by chloramphenicol acetyl-transferase assays in cells transfected with the pFVII-CAT plasmid. HNF-4alpha binding and protein levels were determined by gel shift assays and Western blot. TGF-beta1 and nitric oxide downregulated factor VII mRNA levels by inhibiting its gene promoter activity. This inhibition is caused by a decrease in the DNA binding of HNF-4alpha. TGF-beta1 induces degradation of HNF 4alpha in the proteasome while nitric oxide provokes nitrosylation of cysteine residues in this factor. TGF-beta1 and nitric oxide inhibit HNF-4alpha activity. These findings may explain the loss of liver functions that occurs during fibrosis progression. PMID- 15358162 TI - The differential coupling of oxytocin receptors to uterine contractions in murine estrous cycle. AB - Oxytocin (OT)-induced uterine contractility in different ovulatory phases has not been characterized in mouse. While the uterus was refractory to OT at diestrous phase, numerous contractions were observed at estrous phase. The magnitude of uterine contractile response to OT at estrous phase was larger than that at diestrous phase. There was also significant difference in the pD(2) values for OT between the samples from the two phases. However, the level of OT receptor (OTR) transcripts and proteins was equivalent at the two phases. These results suggest that OT-induced uterine contraction is not only regulated by the quantity of OTR in the non-pregnant mouse uterus. PMID- 15358163 TI - Effect of carbonyl compounds on red blood cells deformability. AB - The effect of Maillard reaction on red blood cells (RBC) deformability was investigated. Exposure of RBC to carbonyl compounds (dl-glyceraldehyde, glyoxal, glycolaldehyde, 3-deoxyglucosone, and d-glucose) leading to Maillard reaction caused a marked decrease in RBC deformability even at 1mM level. The decrease rate depended on the kind of carbonyl compounds, in which both dl-glyceraldehyde and glyoxal significantly decreased the RBC deformability (p<0.05). In addition, the decrease rate also differed among volunteers tested, indicating that the sensitivity against carbonyl compounds varies among them. In order to elucidate the mechanism of the decrease in RBC deformability, RBC was exposed to carbonyl compounds in the presence of aminoguanidine which is the inhibitor of AGE formation in Maillard reactions. Aminoguanidine inhibited the decrease in RBC deformability by dl-glyceraldehyde and glyoxal. When Hb which has a high reactivity with carbonyl compounds was incubated with those carbonyl compounds, dl-glyceraldehyde and glyoxal showed the high reactivity with Hb compared with other carbonyl compounds. These results indicate that Maillard reaction between RBC proteins and carbonyl compounds leads to the decrease in RBC deformability. On the other hand, O(2)(-) generated by carbonyl compounds involved in lowering the deformability only to a negligible level. PMID- 15358164 TI - Dioxin-responsive AHRE-II gene battery: identification by phylogenetic footprinting. AB - We identified a set of genes that respond to dioxins through the recently discovered AHRE-II ("XRE-II") enhancer element. A total of 36 genes containing AHRE-II motifs conserved across human, mouse, and rat gene orthologs were identified by genome-wide transcription-factor binding-site searches and phylogenetic footprinting. Microarray experiments on liver from rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin revealed statistically significant changes in mRNA levels for 13 of these 36 genes after three hours and 15 genes after 19h. The set of responsive genes was functionally characterized by ontological analysis and found to be enriched in ion-channels and transporters. Our identification of 36 putatively AHRE-II-regulated genes highlights the regulatory versatility of the aryl hydrocarbon receptor (AHR) and the ability of the AHR and its dimerization partner, ARNT, to act both as a ligand-activated transcription factor (on AHRE-I) and as a ligand-activated coactivator (on AHRE-II). Collectively, these results demonstrate that the AHRE-II induction mechanism is employed by multiple genes and provide the first categorization of the gene battery of a ligand-activated coactivator. PMID- 15358165 TI - Antiapoptotic effect of ouabain on human umbilical vein endothelial cells. AB - The present study investigates the effect of ouabain on caspase-3 activation in human umbilical vein endothelial cells (HUVEC). Ouabain (EC(50) 20 nM) reduced caspase-3 activity in HUVEC treated for 24h in a medium deprived of fibroblast growth factor (FGF). Incubation for 5h in the absence of both FGF and serum produced an increase in caspase-3 activity that was completely abolished by 100 nM ouabain. Pretreatment with the phosphatidylinositol 3 kinase (PI-3K) inhibitor, wortmannin, prevented the protective effect of ouabain against serum deprivation. Furthermore, Western blotting analysis revealed an increase in phosphorylation of extracellular signal-regulated kinases (ERK-1 and ERK-2) induced by 100nM ouabain in serum-deprived cells. In accord, pretreatment of HUVEC with PD98059, inhibitor of the ERK pathway, abrogated the effect of ouabain. Our results show that ouabain has an antiapoptotic effect on HUVEC through the activation of PI-3K and ERK dependent pathways. PMID- 15358166 TI - Involvement of p38 signaling pathway in interferon-alpha-mediated antiviral activity toward hepatitis C virus. AB - We studied the involvement of the p38 signaling pathway in the interferon (IFN) alpha-mediated antiviral activity toward hepatitis C virus (HCV) using HCV subgenomic replicon cells. When the cells were treated with IFN-alpha in the presence of p38 inhibitor, the suppressive effect of IFN-alpha on replicon RNA was reduced. Inhibition of p38 had almost no influence on phosphorylation of signal transducer and activator transcription factor 1 (STAT1) and interferon stimulatory response element-dependent gene expression after IFN-alpha treatment. This indicates that the anti-HCV activity through p38 may be independent of the Janus kinase-STAT pathway. Treatment with the inhibitor of the mitogen-activated protein kinase-activated protein kinase 2 (MK2) showed the same level of reduction in the IFN-alpha-mediated anti-HCV activity as that with the p38 inhibitor. Thus, MK2 may also be responsible for the anti-HCV activity through p38. In conclusion, the p38-MK2 signaling pathway may be substantially involved in the IFN-alpha-mediated anti-HCV activity. PMID- 15358167 TI - Retention of aggregated LDL by cultured human coronary artery endothelial cells. AB - Aggregated LDL (AgLDL) accumulates within the subendothelial space of developing atherosclerotic lesions. We were interested to learn whether endothelial cells can interact with AgLDL. Incubation of endothelial cells with AgLDL resulted in apparent cholesterol retention. Microscopic examination revealed that cholesterol retention resulted mainly from endothelial cell surface attachment of AgLDL. Little AgLDL entered endothelial cells consistent with the small amount of endothelial cell degradation of AgLDL. Although endothelial cell retention of AgLDL was inhibited by LDL, AgLDL retention was not blocked by lactoferrin, C7 anti-LDL receptor monoclonal antibody, or receptor-associated protein, suggesting that LDL receptor family members did not mediate this retention. Surface retention of AgLDL depended on microtubule function and could be regulated by the protein kinase C activator, PMA. Treatment of endothelial cells with PMA either before or during, but not after incubation with AgLDL, inhibited retention of AgLDL. Our findings show that endothelial cells can retain AgLDL but internalize and metabolize little of this AgLDL. Thus, it is unlikely that endothelial cells can transport AgLDL out of atherosclerotic lesions, but it is likely that retention of AgLDL affects endothelial function. PMID- 15358168 TI - HSF1 blockade-induced tumor thermotolerance abolishment is mediated by JNK dependent caspase-3 activation. AB - We previously blocked the heat shock transcription factor 1 function with a dominant-negative mutant (mHSF1) in breast cancer cell line Bcap37, and found that mHSF1 sensitizes Bcap37 cells to hyperthermia by promoting the apoptotic process. Here we studied the mechanism of this abolishing process and how thermotolerance develops in Bcap37 cells. The results indicated that mHSF1 abolished acquired or intrinsic thermotolerance in Bcap37 cells by enhancing JNK and caspase-3 pathways, two stress-induced apoptotic pathways, after hyperthermia, and interference with either one of them attenuated hyperthermia induced apoptosis. Furthermore, epistasis assay of these two pathways suggested that JNK was upstream of the caspase-3 pathway. Conversely, other hyperthermia induced kinases implicated in cell survival and death, Akt, ERK or p38, did not influence the effect of mHSF1, indicating that these kinases were not implicated in this abolishing process. In addition, we found that the development of acquired thermotolerance of Bcap37 cells was associated with the suppression of JNK activation after mild preheat treatment and was not reduced by Akt, ERK or p38 inhibition. In contrast, the intrinsic thermotolerance of Bcap37 cells was due to the intrinsic high levels of Akt and ERK activities since Akt or ERK inhibition resulted in increased thermosensitivity of Bcap37 cells. Our results suggest that mHSF1 plays a valuable role in the thermotolerance abolishment of Bcap37 cells, which likely contributes to tumor therapy in combination with hyperthermia. PMID- 15358169 TI - Comparative analysis of the proteinase specificity in wild-type and stabilized plasminogen activator inhibitor-1: evidence for contribution of intramolecular flexibility. AB - PAI-1, the physiological inhibitor of tissue-type and urokinase-type plasminogen activator, is a unique member of the serpins as it exists in three distinct conformations: an active inhibitory conformation, a non-inhibitory substrate conformation, and a non-reactive latent conformation. Proline substitution of single residues in the P16-P20 region (situated at the proximal hinge of the reactive site loop) of wild-type PAI-1 (wtPAI-1) and a stabilized PAI-1-variant (PAI-1-stab; N150H, K154T, Q301P, Q319L, and M354I, t(1/2)=150), respectively, resulted in two series of PAI-1-variants with different properties. In wtPAI-1 only substitution at P18 resulted in a pronounced u-PA specificity and substrate behaviour towards t-PA. In contrast, in PAI-1-stab substitution at either P18, P19 or P20 resulted in a u-PA specificity and a significantly increased substrate behaviour towards t-PA and u-PA. Importantly, analysis of the kinetics of inhibition did not reveal any differences in the second-order rate constants of inhibition (k approximately 10(7)M(-1)s(-1)). The pronounced differences observed for identical mutations in wtPAI-1 vs PAI-1-stab demonstrate that not merely the sequence of the reactive loop but also intramolecular interactions between the hF/s3A-loop and the main part of the molecule govern the functional and conformational behaviour of PAI-1. PMID- 15358170 TI - Correspondence regarding M.V. Kim et al. "Some properties of human small heat shock protein Hsp22 (H11 or HspB8)". PMID- 15358171 TI - A hepatocellular carcinoma cell line producing mature hepatitis B viral particles. AB - Current in vitro models for hepatitis B virus (HBV) are based on human hepatoblastoma cell lines transfected with HBV genome. The objective of this work was to develop an in vitro, hepatocellular carcinoma (HCC)-based system supporting HBV full replication and producing mature viral particles. The FLC4 human HCC cell line was stably transfected with a plasmid carrying a head-to-tail dimer of the adwHBV genome. One of the clones, FLC4A10II, exhibited prolonged expression of HBV, as was demonstrated by secreted levels of HBsAg, HBeAg, and HBV DNA in the culture medium of the growing cells. Furthermore, the cells produced HBV particles that were detected by a cesium chloride density gradient performed on the culture medium. Analysis by Southern blot revealed that HBV DNA has integrated into the FLC4A10II cell genome. The presence of HBV in the FLC4A10II cells did not cause alterations in cell morphology and the cells continued to resemble mature hepatocytes. They do exhibit a high mitotic activity. The new HBV stably transfected cell line, FLC4A10II, can serve as an important tool for further exploration of HBV host-pathogen interaction, viral life cycle, and for assessing new antiviral agents. PMID- 15358172 TI - Lineage-specific cell disruption in living mice by Cre-mediated expression of diphtheria toxin A chain. AB - We have established a transgenic mouse line in which floxed neomycin resistant cassette was inserted between the CAG promoter and EGFP. When these transgenic mice were mated with Cre-expressing transgenic animals, the offspring obtained were fluorescent green. We then established a transgenic mouse line in which EGFP in the above construct was replaced by diphtheria toxin A chain (DT). When the latter transgenic mice were mated with mice expressing Cre restricted to germ cells, we obtained healthy but sterile offspring due to a disruption of germ line cells by DT expression. We predict that this strategy will be useful for the construction of new animal models for human diseases, featuring a variety of missing cell lineages produced by disruption with DT. PMID- 15358173 TI - Construction of a novel human artificial chromosome vector for gene delivery. AB - Potential problems of conventional transgenes include insertional disruption of the host genome and unpredictable, irreproducible expression of the transgene by random integration. Alternatively, human artificial chromosomes (HACs) can circumvent some of the problems. Although several HACs were generated and their mitotic stability was assessed, a practical way for introducing exogenous genes by the HACs has yet to be explored. In this study, we developed a novel HAC from sequence-ready human chromosome 21 by telomere-directed chromosome truncation and added a loxP sequence for site-specific insertion of circular DNA by the Cre/loxP system. This 21HAC vector, delivered to a human cell line HT1080 by microcell fusion, bound centromere proteins A, B, and C and was mitotically stable during long-term culture without selection. The EGFP gene inserted in the HAC vector expressed persistently. These results suggest that the HAC vector provides useful system for functional studies of genes in isogenic cell lines. PMID- 15358174 TI - TAP/NXF1, the primary mRNA export receptor, specifically interacts with a neuronal RNA-binding protein HuD. AB - Hu proteins are RNA-binding proteins that are implicated in the control of stabilization, nuclear export, and/or translation of specific mRNAs with AU-rich elements (AREs) in the 3'-untranslated region. Three neuron-specific Hu proteins (HuD, HuB, and HuC), but not a ubiquitously expressed Hu protein HuR, have an activity to induce neurite outgrowth when they are overexpressed in a rat neuronal cell line PC12. Here we show that TAP/NXF1, the primary export receptor for the bulk mRNA, is a specific binding partner for HuD. In vitro binding experiments using recombinant proteins revealed that the interaction between TAP and HuD is direct and that HuD can form a ternary complex together with both TAP and RNA. Interestingly, HuR does not interact with TAP. These results suggest that HuD acts as a novel adaptor protein to recruit TAP for efficient export of ARE-containing mRNAs in neuronal cells. PMID- 15358175 TI - Nitric oxide induces BNIP3 expression that causes cell death in macrophages. AB - Nitric oxide (NO) is involved in many physiological processes and also causes pathological effects by inducing apoptosis. It can enhance or suppress apoptosis depending on its concentration and the cell type involved. In this report, we used cDNA microarray analysis to show that SNAP, an NO donor, strongly induces Bcl-2/adenovirus E1B 19kDa-interacting protein 3 (BNIP3) in macrophages. BNIP3 is a mitochondrial pro-apoptotic protein that contains a Bcl-2 homology 3 domain and a COOH-terminal transmembrane (TM) domain. Macrophages activated by LPS/IFN-gamma produce nitric oxide synthase 2 (NOS2) and release endogenous NO. Expression of BNIP3 was also induced in macrophages by LPS/IFN-gamma, and the induction was blocked by a NOS2 inhibitor, S-methyl-isothiourea. Peritoneal macrophages from NOS2-null mice failed to produce BNIP3 in response to LPS/IFN-gamma. We conclude that BNIP3 expression in macrophages is controlled by the intracellular level of nitric oxide. Overexpression of BNIP3 but not of BNIP3 deltaTM, a BNIP3 mutant without the TM domain and C-terminal tail, led to apoptosis of the cells. Promoter analysis showed that the region between -281 and -1 of the 5'-upstream enhancer region of murine BNIP3 was sufficient for NO-dependent expression of BNIP3. PMID- 15358176 TI - The human Duffy antigen binds selected inflammatory but not homeostatic chemokines. AB - The aim of the study was to compare the ability of the human Duffy antigen to bind homeostatic and inflammatory chemokines. Homeostatic chemokines did not bind to the Duffy antigen on erythrocytes with high affinity. In contrast, 60% of inflammatory chemokines bound strongly to Duffy, with no obvious preference for CXC or CC classes. It was investigated if this binding profile was reflected in the binding pattern of endothelial cells. Two examples of homeostatic (125I CXCL12 and 125I-CCL21) and inflammatory (125I-CXCL8 and 125I-CCL5) chemokines were incubated with human synovia. In agreement with the erythrocyte binding data, intense specific signals for CXCL8 and CCL5 binding were found on endothelial cells, whereas CXCL12 and CCL21 showed only weak binding to these cells. Our study provides evidence that the human Duffy antigen binds selected inflammatory, but not homeostatic, chemokines and that this binding pattern is reflected by endothelial cells within inflamed and non-inflamed tissue. PMID- 15358177 TI - Determination of sensitive positions to mutations in human p53 protein. AB - Over last several years, we demonstrated that the mutations are more likely to occur at randomly unpredictable amino acid pairs in a protein. We therefore can in principle predict the amino acid pairs sensitive to the future mutations in a protein. However, we still need to predict the positions at which the sensitive amino acid pairs are located in a protein. In this study, we use a probabilistic approach to analyze the effect of 191 mutations in human p53 protein and can approximately estimate the sensitive positions to mutations in human p53 protein. PMID- 15358178 TI - Promoter polymorphism in fibroblast growth factor 1 gene increases risk of definite Alzheimer's disease. AB - Fibroblast growth factor 1 (FGF1, also known as acidic FGF) protects selective neuronal populations against neurotoxic effects such as those in Alzheimer's disease (AD) and HIV encephalitis. The FGF1 gene is therefore a strong candidate gene for AD. Using the promoter polymorphism of the FGF1 gene, we examined the relationship between AD and the FGF1 and apolipoprotein E (APOE) genes in 100 Japanese autopsy-confirmed late-onset AD patients and 106 age-matched non demented controls. The promoter polymorphism (-1385 A/G) was significantly associated with AD risk. The odds ratio for AD associated with the GG vs non-GG genotype was 2.02 (95% CI = 1.16-3.52), while that of s4 vs non-l4 in APOE4 gene was 5.19 (95% CI = 2.68-10.1). The odds ratio for APOEP4 and FGF1 GG carriers was 20.5 (95% CI = 6.88-60.9). The results showed that the FGF1 gene is associated with autopsy-confirmed AD. PMID- 15358179 TI - Zinc binding agonist effect on the recognition of the beta-amyloid (4-10) epitope by anti-beta-amyloid antibodies. AB - Amyloid plaques associated to Alzheimer's disease present a high content of zinc ions. We previously showed that the N-terminal region of the amyloid peptide Abeta constitutes an autonomous zinc-binding domain. This region encompasses the previously identified epitope Abeta(4-10) targeted by antibodies capable to reduce amyloid deposition, but the influence of Abeta/Zn binding on the epitope recognition remains unknown. We demonstrate here the effect of Zn2+ ions on the recognition of peptides sharing the sequence of the Abeta N-terminal domain, by two monoclonal antibodies recognizing the beta-amyloid(4-10) epitope. The presence of Zn2+, but not of other cations, increased the recognition of the (1 16) peptide, while it was without effect on the recognition of the (1-10) peptide. These findings show a zinc-induced conformational change of the (1-16)-N terminal region of AP3, which results in a better accessibility of the Abeta(4 10) epitope to the anti-Abeta antibodies, and suggest a role of zinc in epitope based vaccination approaches. PMID- 15358180 TI - A novel human dendritic cell-derived C1r-like serine protease analog inhibits complement-mediated cytotoxicity. AB - Trypsin-like serine proteases are involved in diverse biological processes such as complement activation, tissue remodeling, cellular migration, tumor invasion, and metastasis. Here we report a novel human C1r-like serine protease analog, CLSPa, derived from dendritic cells (DC). The 487-residue CLSPa protein contains a CUB domain and a serine protease domain, possessing characteristic catalytic triad but lacking typical activation/cleavage sequence. It shares great homology with complement C1r/C1s and mannose-associated serine proteases. CLSPa mRNA is widely expressed, especially abundant in placenta, liver, kidney, pancreas, and myeloid cells, which are a major resources of serine proteases. Upon stimulation by agonistic anti-CD40 Ab, TNF-alpha, or LPS, CLSPa mRNA expression was significantly up-regulated in monocytic cells and monocyte-derived immature DC. When overexpressed in 293T cells, CLSPa protein was synthesized into the culture supernatants as a secretory protein, which had an inhibitory effect on complement mediated cytotoxicity to antibody-sensitized erythrocytes. However, CLSPa itself possesses little protease activity, but it plays an inhibitory role in other active protease catalytic processes. The identification of human CLSPa as a novel Clr-like protein might facilitate future investigation of the regulatory mechanism of CLSPa in complement pathways during inflammation. PMID- 15358181 TI - Synthetic curcumin analogs inhibit activator protein-1 transcription and tumor induced angiogenesis. AB - In a previous study, we observed that some synthetic curcumin analogs inhibited complex formations between Fos-Jun heterodimer and activator protein-1 (AP-1) DNA. These curcumin analogs have been observed to repress the AP-1 transcription in AP-1-transfected cells and they also inhibited the increased expression of Jun/AP-1 protein by 12-O-tetradecanoylphorbol-13-acetate (TPA) in the same cells. After the AP-1 inhibition by curcumin analogs in TPA-treated HT-1080 human fibrosarcoma cells, a decrease in mRNA expression of c-jun and MMP3 (stromelysin 1) has been observed. We also observed that curcumin analogs down-regulated the expression of MMP-9 (gelatinase-B), correlating with cellular invasion and migration in conditions such as tumor invasion and metastasis, through the electrophoretic mobility shift assay and gelatin zymography methods. Curcumin analogs showed an inhibitory effect on angiogenesis by various test methods including chicken chorioallantoic membrane assay, wound migration assay, invasion assay, and tube formation assay. Through the reverse transcriptase-polymerase chain reaction experiment, we confirmed that curcumin analogs down-regulated the expression of angiogenesis-associated genes, VEGF and MMP-9. PMID- 15358182 TI - Activators of PPARgamma antagonize protection of cardiac myocytes by endothelin 1. AB - Endothelin-1 (ET-1) is a potent survival factor against myocardial cell apoptosis. This anti-apoptotic effect of ET-1 is mediated in part through calcineurin/NFATc-dependent induction of bcl-2 expression. Since it has been reported that peroxisome proliferator-activated receptor-gamma (PPARgamma) interacts with NFATc, we investigated the effects of PPARgamma ligands on anti apoptotic effects of ET-1 in cardiac myocytes. In primary cardiac myocytes from neonatal rats, administration of PPARgamma activators (15-deoxy-delta12,14 prostaglandin J2 and troglitazone) attenuated the anti-apoptotic effects of ET-1. These activators abolished the ET-1-stimulated increase in bcl-2 expression and in binding of cardiac NFATc to the bcl-2 NFAT site. These findings demonstrate that activators of PPARgamma perturb the anti-apoptotic effects of ET-1 in cardiac myocytes and that this perturbation is, in part, based on functional transcriptional cross-talk between NFATc and PPARgamma. PMID- 15358183 TI - Afadin- and alpha-actinin-binding protein ADIP directly binds beta'-COP, a subunit of the coatomer complex. AB - Afadin DIL domain-interacting protein (ADIP) is a novel protein that binds both afadin and alpha-actinin and localizes at adherens junctions, which are formed by nectins and cadherins, cell-cell adhesion molecules. Afadin is an actin filament (F-actin)-binding protein which connects nectins to the actin cytoskeleton. alpha Actinin is another F-actin-binding protein that is indirectly associated with cadherins through the catenin complex. ADIP is at least partly involved in the physical association of nectins and cadherins. We show here that ADIP furthermore binds beta'-COP, a subunit of the coatomer complex. ADIP co-localizes with beta' COP at the Golgi complex in Madin Darby canine kidney and normal rat kidney cells. These results suggest that ADIP is involved in vesicle trafficking from the Golgi to the endoplasmic reticulum and through the Golgi complex by interacting with the coatomer complex. PMID- 15358184 TI - Characterization of Smubp-2 as a mouse mammary tumor virus promoter-binding protein. AB - A cDNA encoding a rat Smubp-2 has been cloned from a lambdagt11 library by South Western blot screening using a 50-bp tannic acid responsive element [J. Biol. Chem. 273 (1998) 12499] of the mouse mammary tumor virus (MMTV) promoter region as a probe. The full-length cDNA encodes a protein with a predicted size of 108 kDa. Northern blot analysis revealed that the gene expression of Smubp-2 is comparatively high in testis, moderate in brain, and low in other tissues. The recombinant Smubp-2 protein was expressed as a GST- or Trx-fusion protein in Escherichia coli and purified by affinity column chromatography. Gel mobility shift competition analysis indicated that the recombinant Smubp-2 protein binds to region II (containing the ACTG-motif) in the 50-bp element in the MMTV promoter. A transient transfection assay of the Smubp-2 expression vector with MMTV promoter-containing Luciferase (Luc) reporter plasmids into mouse cells suggested that Smubp-2 is a negative transcription factor. Furthermore, the MMTV promoter activity was suppressed in cells expressing high levels of Smubp-2. Insertion of the 50-bp element upstream of the SV40 promoter negatively responded to the induced expression of Smubp-2. These results suggest that the negative transcriptional effect of Smubp-2 arises from its binding to the 50-bp element located in the MMTV promoter region. PMID- 15358185 TI - Gene silencing using a heat-inducible RNAi system in Arabidopsis. AB - Controlling gene expression during plant development is an efficient tool to explore gene function. In this paper, we describe a gene expression system driven by a heat-shock gene promoter (HSP18.2), to trigger the expression of an intron containing inverted-repeat. RNA interference became a powerful way for gene functional analysis by reverse genetic approaches. However, constitutive gene silencing cannot be used with genes involved in fundamental processes such as embryo viability. Inducible promoters provide an alternative approach for temporal and spatial gene expression control and we described here a new system, complementary to those using chemical gene inducers. To evaluate the efficiency of this system, RNA corresponding to the phytoene desaturase gene of Arabidopsis thaliana was used as a reporter gene in transgenic plants and a comparative study was performed using either the CaMV35S constitutive promoter or the HSP18.2 inducible promoter. PMID- 15358186 TI - "Teaching old drugs to kill new bugs": structure-based discovery of anti-SARS drugs. AB - Severe acute respiratory syndrome (SARS) main protease or 3C-like protease (3CLpro) is essential for the propagation of the coronaviral life cycle and is regarded as one of the main targets for structure-based anti-SARS drug design. It is an attractive approach to find new uses for old drugs as they have already been through extensive clinical testing and could easily be accelerated for clinical approval. Briefly, we performed virtual screening of a database of small molecules against SARS 3CLpro, analyzed inhibitor-protease complexes, and identified several covalent and non-covalent inhibitors. Several old drugs that bind to SARS 3CLpro active site were selected and in silico derivatized to generate covalent irreversible inhibitors with enhanced affinity. Furthermore, we show that pharmacophores derived from clusters of compounds resulting out of virtual screening could be useful probes for future structure-activity relationship studies (SARs) and fine-tune the lead molecules identified. PMID- 15358187 TI - Interference of 7,8-dihydroneopterin with peroxynitrite-mediated reactions. AB - By in vitro studies 7,8-dihydroneopterin, which is secreted by macrophages stimulated by interferon-gamma, was reported to be a radical scavenger as well as a prooxidative agent depending on the experimental settings. In this study, we investigated the interference of 7,8-dihydroneopterin with peroxynitrite mediated reactions by different analytical procedures. Luminol chemiluminescence and oxidation of the spin probe 1-hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine induced by peroxynitrite were inhibited by 7,8-dihydroneopterin. On the other hand, we found that 7,8-dihydroneopterin very efficiently inhibits nitration of tyrosine by peroxynitrite. Hydroxylation, however, was rather enhanced than inhibited, suggesting that 7,8-dihydroneopterin reacts in quite different manner with the intermediates generated from peroxynitrite. We provide the first evidence that a pterin radical is formed from a dihydropterin using EPR spectroscopy and 2,2,4-trimethyl-2H-imidazole-1-oxide as a spin trap. We conclude that 7,8-dihydroneopterin while being a weak scavenger of superoxide acts as a very efficient inhibitor of tyrosine nitration induced by peroxynitrite. PMID- 15358188 TI - Senescence marker protein-30 regulates Akt activity and contributes to cell survival in Hep G2 cells. AB - Senescence marker protein-30 (SMP30) is highly expressed in cytosol of hepatocytes, and its amount decreases with aging. Human hepatocellular carcinoma cell line was transfected with pcDNA3/SMP30 (SMP30 transfectants), or as a control with pcDNA3 (mock transfectants). When cells were exposed to 20 ng/ml tumor necrosis factor-alpha (TNF-alpha) plus 10 ng/ml actinomycin D (Act-D) for 15 h, the viability of cells was decreased in both SMP30 and mock transfectants. However, the viability of cells was threefold higher in SMP30 transfectants than mock transfectants. Cell death was confirmed as apoptosis by TUNEL assay. The presence of trifluoperazine, a calmodulin (CaM) inhibitor, attenuated anti apoptotic effect of SMP30 in both transfectants, but the effect was more prominent in SMP30 transfectants. Western blot analyses revealed that Akt, which acts as a survival factor in cells, was activated in SMP30, but not mock, transfectants either in the presence or absence of TNF-alpha plus Act-D. Further, trifluoperazine inhibited Akt activation in SMP30 transfectants. We therefore propose that interplay between CaM and SMP30 regulates Akt activity, and thus SMP30 acts as a survival factor in hepatocytes. PMID- 15358189 TI - Computational analysis of responsible dipeptides for optimum pH in G/11 xylanase. AB - A bioinformatics method was used to search the responsible dipeptides for optimum pH in the G/11 xylanase, for dipeptides can provide position information of the related residues for rational protein design. The responsible dipeptides were found as negative YS and positive SY, GR, MR, and KR. The minimum and maximum optimum pH was calculated as 2.33 and 14.29, respectively. Compared with the known crystal structures of the G/11 xylanase, YS was found mostly in the turn area of beta-strands of S/T surface; and SY was found in the inner part of beta strands of the S/T area near to the active site of proton donor; and the GR, MR, and KR in the coil region connecting "finger" to the alpha-helix. The result clearly explained the success of shifting of pH 0.5 U to alkaline by the introduction of arginines into S/T area of a xylanase. The result would be useful for xylanase engineering, and the adaptation mechanism to high alkaline was also discussed. PMID- 15358190 TI - Trypanosoma cruzi: TcRAB7 protein is localized at the Golgi apparatus in epimastigotes. AB - In mammalian cells, the Rab7 protein is a key element of late endocytic membrane traffic. Several results suggest that it is involved in the transport from early to late endosome or from late endosome to lysosome. We have previously characterized a Rab7 gene homologue (TcRAB7) in Trypanosoma cruzi. Now, using an affinity-purified antibody specific to TcRAB7 protein we have determined that it is localized at the Golgi apparatus of the parasite. Our results indicate that the T. cruzi Rab7 homologue may function in a different route than its counterparts in mammalian cells. PMID- 15358191 TI - Cytotoxicity of the novel anti-cancer drug rViscumin depends on HER-2 levels in SKOV-3 cells. AB - rViscumin is a recombinant mistletoe lectin under clinical investigation as new anti-cancer drug. The relationship between oncogene, e.g., HER-2/neu (c-erbB2) receptor activation and tumor cell chemosensitivity, is of considerable importance to better predict the response to chemotherapy. Here, we analyze the cellular and molecular effects of HER-2 expression on rViscumin chemotoxicity in SKOV-3 cells. We show that selective depletion of HER-2 by ribozyme-targeting markedly decreases cellular sensitivity towards rViscumin. These findings are confirmed by treatment with the well-established inhibitory HER-2 antibody trastuzumab (Herceptin). Using clonal ribozyme-transfected cell lines, we establish a 'HER-2 gene dose' dependence of rViscumin cytotoxicity, which is due to differential induction of apoptosis and is not mediated by cell cycle alterations or altered cellular rViscumin binding/internalization. We further demonstrate an rViscumin-mediated, HER-2-dependent down-regulation of bcl-2 and the dose-dependent activation of members of the MAPK family, p42/44, SAPK/JNK, and p38, but not of caspases-3 and -7. PMID- 15358192 TI - Antisense strategy against PDGF B-chain proves effective in preventing experimental liver fibrogenesis. AB - Hepatic stellate cells (HSCs) and transdifferentiated myofibroblasts are the principal producers of excessive extracellular matrix in liver fibrosis and cirrhosis. Activation of HSC is regulated by several cytokines and growth factors, including platelet-derived growth factor B-chain (PDGF-B), a potent mitogen for HSC, and overexpressed during hepatic fibrogenesis. Previous studies showed that MAPK and phosphatidylinositol 3' kinase are key signaling pathways involved in PDGF-induced stimulation of HSC. Based on the involvement of PDGF-B in fibrogenesis, reducing ligand stimulation of proliferative cytokine- or growth factor receptors interfering with receptor signaling therefore presents an interesting strategy for hepatic fibrosis prevention or interruption. We therefore generated an adenoviral vector serotype 5 (Ad5) expressing an antisense mRNA of the PDGF B-chain (Ad5-CMV-asPDGF) for application in an experimentally induced liver fibrogenesis model. The transgene clearly showed the ability to down-regulate endogenous PDGF B-chain and PDGFRbeta mRNA in culture-activated HSC and rat livers. The asPDGF mRNA also attenuates experimental liver fibrogenesis indicated by reduced levels of alpha-SMA and collagen type I expression. PMID- 15358193 TI - The in vivo tyrosine phosphorylation level of yeast immunophilin Fpr3 is influenced by the LMW-PTP Ltp1. AB - Tyr-phosphorylation in Saccharomyces cerevisiae is essential in controlling the activity of MAP kinase regulating mating, pseudohyphal growth, and cell wall biosynthesis. Yeast serves as a model system for studying the biological function of many protein kinases and PTPs. Two LMW-PTP from yeast have been cloned, namely, Ltp1 from S. cerevisiae and Stp1 from Schizosaccharomyces pombe. The sequences of both enzymes are relatively similar to those of the mammalian LMW PTP. Recently we showed that the yeast immunophilin Fpr3 interacts with Stp1 and its dephosphorylated state induces a growth defective phenotype. Here we show the phosphatase activity of Ltp1 on Fpr3 and we demonstrated that Tyr 184 is the residue phosphorylated on in vivo Fpr3. We also described the marked activation of Ltp1 by adenine in S. cerevisiae proteome and determined in vivo the influence of tyrosine phosphorylation on Fpr3 localization. PMID- 15358194 TI - Detection of the novel autoantibody (anti-UACA antibody) in patients with Graves' disease. AB - Uveal autoantigen with coiled coil domains and ankyrin repeats (UACA) is an autoantigen in patients with panuveitis such as Vogt-Koyanagi-Harada disease. The prevalence of IgG anti-UACA antibodies in patients with uveitis is significantly higher than healthy controls, suggesting its potential role as an autoantigen. Originally, UACA was cloned from dog thyroid tissue following TSH stimulation. So, we presumed UACA could be a novel autoantigen in autoimmune thyroid diseases. We measured serum anti-UACA antibody titer using ELISA in patients with autoimmune thyroid diseases (Graves' disease, Hashimoto's thyroiditis, subacute thyroiditis, and silent thyroiditis). The prevalence of anti-UACA antibodies in Graves' disease group was significantly higher than that in healthy group (15% vs. 0%). Moreover, the prevalence of anti-UACA antibodies in Graves' ophthalmopathy was significantly higher than that in Graves' patients without ophthalmopathy (29% vs. 11%). Especially, 75% of severe ocular myopathy cases showed high UACA titer. Immunohistochemical analysis revealed that UACA protein is expressed in eye muscles as well as human thyroid follicular cells. Taken together, UACA is a novel candidate for eye muscle autoantigens in thyroid associated ophthalmopathy. PMID- 15358195 TI - Modulation of Janus kinase 2 by p53 in ovarian cancer cells. AB - The constitutive activation of the Janus kinase 2 (JAK2) and mutation of the p53 tumor suppressor are both detected in human cancer. We examined the potential regulation of JAK2 phosphorylation by wild-type (wt) p53 in human ovarian cancer cell lines, Caov-3 and MDAH2774, which harbor mutant form of p53 tumor suppressor gene and high levels of phosphorylated JAK2. The wt p53 gene was re-introduced into the cells using an adenovirus vector. In addition to wt p53, mutant p53 22/23, mutant p53-175, and NCV (negative control virus) were introduced into the cells in the control groups. Expression of wt p53, but not that of p53-175 mutant, diminished JAK2 tyrosine phosphorylation in MDAH2774 and Caov-3 cell lines. Expression of wt p53 or p53 22/23 mutant did not cause a reduction in the phosphorylation of unrelated protein kinases, ERK1 and ERK2 (ERK1/2). The inhibition of JAK2 tyrosine phosphorylation can be reversed by tyrosine phosphatase inhibitor, sodium orthovanadate. Protein tyrosine phosphatase 1-B levels increased with introduction of wt p53 and may be involved in the dephosphorylation of JAK2. These findings present a possible p53-dependent cellular process of modulating JAK2 tyrosine phosphorylation in ovarian cancer cell lines. PMID- 15358196 TI - MUC1 tyrosine phosphorylation activates the extracellular signal-regulated kinase. AB - MUC1 is a transmembrane glycoprotein expressed on the apical surface of epithelial cells and exhibiting structural features characteristic of receptors for cytokines and growth factors. Its intracellular cytoplasmic tail (CT) contains multiple amino acid sequence motifs that, once phosphorylated, serve as docking sites for SH2 domain-containing proteins mediating signal transduction. Most studies examining MUC1 signaling have focused on cancer cells where MUC1 is overexpressed, aberrantly glycosylated, and constitutively phosphorylated. No studies have determined the signaling pathways activated in response to stimulation of its ectodomain. To better understand the signaling mechanisms of MUC1, we stably transfected HEK293 cells with an expression plasmid encoding a chimeric protein consisting of the extracellular and transmembrane domains of CD8 and the MUC1 CT (CD8/MUC1). Extracellular treatment of HEK293-CD8/MUC1 cells with CD8 antibody induced intracellular Tyr phosphorylation of the MUC1 CT and activated ERK1/2, but not the p38, SAPK/JNK, or ERK5 MAP kinases. Moreover, phosphorylation of ERK1/2 was completely blocked using a CT deletion mutant or a mutant construct in which all Tyr residues in the CT were changed to Phe. These results establish that Tyr phosphorylation of the MUC1 CT is required for activation of a downstream ERK1/2 pathway. PMID- 15358197 TI - Age-dependent methylation of ESR1 gene in prostate cancer. AB - The incidence of prostate cancer increases dramatically with age and the mechanism underlying this association is unclear. Age-dependent methylation of estrogen receptor alpha (ESR1) gene has been previously implicated in other cancerous and benign diseases. We evaluated the age-dependent methylation of ESR1 in prostate cancer. The methylation status of ESR1 in 83 prostate cancer samples from patients aged 49 to 77 years (mean age at 67.4 years) was examined using the bisulfite genomic sequencing technique. The samples were divided into three age groups: men aged 60 years and under (n = 14), men aged 61-70 years (n = 40), and men aged over 70 years (n = 29). Overall, ESR1 promoter methylation was detected in 54 out of 83 (65.1%) prostate samples. The methylation rate of ESR1 increased dramatically with age from 50.0% in patients aged 60 years and under to 89.7% for patients aged 70 years and over. Logistic regression analyses revealed that age and Gleason score were the only variables that affect incidence of ESR1 methylation; other clinical factors such as prostate-specific antigen level and clinical stage did not. We also calculated ESR1 methylation density (the percentage of methylated CpGs among all CpGs within the analyzed region) and severity (the percentage of methylated CpG alleles) for each sample analyzed. Multiple regression analyses showed a positive correlation between age and methylation density (beta, 0.35; P, 0.012; 95% CI, 0.26-2.01); while Gleason score was positively associated with methylation severity (beta, 0.45; P, 0.018; 95% CI, 1.04-4.26). These findings suggest that methylation of ESR1 is both age dependent and tumor differentiation-dependent and age-dependent methylation of ESR1 may represent a mechanism linking aging and prostate cancer. PMID- 15358198 TI - Genistein induces Ca2+ -mediated, calpain/caspase-12-dependent apoptosis in breast cancer cells. AB - Genistein, a soy-derived isoflavone, has been suggested for breast cancer prevention; however, use of soy products for this purpose remains controversial. Genistein has been reported to regulate growth of tumor cells, although the involved molecular mechanisms are not defined. Here we report that genistein induces apoptosis in breast cancer cells via activation of the Ca2+ -dependent proapoptotic proteases, mu-calpain, and caspase-12. The treatment of MCF-7 breast cancer cells with genistein induced a sustained increase in concentration of intracellular Ca2+ resulting from depletion of the endoplasmic reticulum Ca2+ stores. This increase in Ca2+ was associated with activation of mu-calpain and caspase-12, as evaluated with the calpain and caspase-12 substrates and antibodies to active (cleaved) forms of the enzymes. Selective inhibition of Ca2+ binding sites of mu-calpain, forced increase of the cytosolic Ca2+ buffering capacity, and caspase inhibition decreased apoptotic indices in the genistein treated cells. Our results suggest that Ca2+ -dependent proteases are potential targets for genistein in breast cancer cells and that the cellular Ca2+ regulatory activity of genistein underlies its apoptotic mechanism. PMID- 15358199 TI - Distribution of a single nucleotide polymorphism on codon 211 of the androgen receptor gene and its correlation with human renal cell cancer in Japanese patients. AB - The human androgen receptor (AR) gene contains a single nucleotide polymorphism (SNP) on codon 211 and two polymorphic short tandem repeats of CAG and GGC in the N-terminal domain that may influence transcription efficiency of AR gene. We previously reported that the lengths of the CAG and GGC repeats are inversely and linearly related to AR activity and associated with several cancers. However, little is known about this SNP on codon 211 of the AR gene in human renal cell cancer. The cause of renal cell cancer is not well understood although several factors such as chemical carcinogens and hormones have been implicated. AR has been identified in human and animal renal cell cancer. We hypothesize that the SNP on codon 211 is associated with human renal cell cancer. To test this hypothesis, the genetic distribution of the SNP on codon 211 of AR gene was investigated in renal cell cancer patients (211 cases) and healthy controls (200 cases). The allelic and genotypic distributions were determined by PCR-RFLP and direct DNA sequencing techniques. The chi2 test for these data revealed that the distribution of this SNP was not different between renal cell cancer patients as compared to normal healthy controls. The findings suggest that the SNP on codon 211 in the AR gene may not have an important role in the carcinogenesis of human renal cell cancers. PMID- 15358200 TI - Bacteriostatic properties of biomatrices against common orthopaedic pathogens. AB - Tissue-engineered grafts for tissue regeneration include either mature or progenitor cells seeded onto biomatrices that provide shape and support for developing tissue. Popular biomaterials used in orthopaedic surgery include collagen type I, hyaluronic acid, hydroxyapatite, and polylactic polyglycolic acid (PLGA). Biomatrices with bacteriostatic properties may be beneficial in promoting tissue-engineered graft survival in patients susceptible to infection. We evaluated the bacteriostatic effects of these biomaterials on the growth of the four most common orthopaedic bacterial pathogens: Staphylococcus aureus, Staphylococcus epidermidis, beta-hemolytic Streptococcus, and Pseudomonas aeruginosa. Hyaluronic acid demonstrated the largest bacteriostatic effect on these pathogens by inhibiting bacterial growth by an average of 76.8% (p = 0.0005). Hydroxyapatite and collagen inhibited growth on average by 49.7% (p = 0.011) and 37.5% (p = 0.102), respectively. PLGA exhibited the least bacteriostasis with an average inhibition of 9.8% (NS) and actually accelerated the growth of beta-hemolytic Streptococcus and P. aeruginosa. PMID- 15358201 TI - The cardioprotective effect of urocortin during ischaemia/reperfusion involves the prevention of mitochondrial damage. AB - We have previously shown, using Affymetrix gene chip technology, that urocortin induces the expression of several diverse genes in cardiac myocytes. An ATP sensitive inwardly rectifying potassium channel, Katp (Kir6.1), the enzyme calcium independent phospholipase A2 (iPLA2), and protein kinase C epsilon (PKCepsilon) and that these genes are involved in the cardioprotective mechanism of action of urocortin. Here we demonstrate that these gene products are localized to cardiac myocyte mitochondria and for the first time show that urocortin protects cardiac myocytes from ischaemia/reperfusion induced cell death by preventing mitochondrial damage. Using pharmacological agents to Katp channels and iPLA2 and synthetic peptide inhibitors of PKCepsilon, we go on to demonstrate that these three gene products are involved in the urocortin induced protection of cardiac myocyte mitochondria. These proteins may interact at the mitochondria to produce the protective effect. PMID- 15358202 TI - The role of Xenopus frizzled-8 in pronephric development. AB - Vertebrates use two or three forms of kidney successively during development and the nephric duct is essential for this succession of kidney induction. While transcripts of many Wnt ligands and Wnt receptor Frizzled genes have been localized in developing kidney, the relationship between Wnt signaling and nephric duct development remains unknown. This study investigated the role of Xenopus frizzled-8 (Xfz8) in pronephros development. Translational inhibition of Xfz8 caused a significant reduction in the staining of a duct-specific antibody, but did not affect the expression of early pronephric maker genes in the duct region. Defects in pronephric tubule branching were also observed following inhibition of Xfz8. Histological analysis revealed that the Xfz8-inhibited cells failed to form a normal epithelium structure. These results suggest that Xfz8 is involved in the process of normal epithelium formation in the developing pronephric duct and tubules after specification. PMID- 15358203 TI - Nek2A specifies the centrosomal localization of Erk2. AB - Nek2A is a cell-cycle-regulated protein kinase that localizes to the centrosome and kinetochore. Our recent studies provide a link between Nek2A and spindle checkpoint signaling [J. Biol. Chem. 279 (2004) 20049]. Extracellular signal regulated kinase 2 (Erk2) is an important kinase, which belongs to mitogen activating protein (MAP) kinase family. Here we demonstrated that Nek2A binds specifically to Erk2. Erk2 interacts with Nek2A via a conserved Erk2 docking site located to the C-terminus of Nek2A. Our studies indicate this docking site is essential and sufficient for a direct Nek2A-Erk2 interaction. In addition, our immunocytochemical studies show that Nek2A and Erk2 are co-localized to centrosome. Significantly, elimination of Nek2A by RNA interference delocalized Erk2 from its centrosomal location, while inhibition of Erk2 kinase activity did not affect the localization of Nek2A in centrosome. We propose that Erk2 links extracellular signaling to centrosome dynamics by Nek2A. PMID- 15358204 TI - Zebrafish heparin-binding neurotrophic factor enhances neurite outgrowth during its development. AB - Heparin-binding neurotrophic factor (HBNF) is a secreted heparin-binding protein containing highly basic and cysteine-rich amino acid residues. In this study, we cloned the full-length HBNF cDNA from zebrafish and determined its genomic structure by bioinformatics analysis. Zebrafish HBNF gene is composed of five exons and four introns spanning approximately 82kb. RT-PCR analysis revealed that zebrafish HBNF transcript was highly expressed in adult brain and intestine tissues while less in other tissues. During embryogenesis, zebrafish HBNF transcript was observed to be moderately expressed at earlier stages with a gradual decline. Higher expression level was observed after hatching and maintaining this level into adulthood. The overall amino acid sequence of zebrafish HBNF shows 60% identity to human HBNF, but with approximately 40% identity to other midkine proteins. Like mammalian homolog, zebrafish HBNF could induce significant neurite outgrowth in PC12 cells without NGF stimulation. In addition, zebrafish HBNF was able to enhance extensive neurite outgrowth in zebrafish embryos during embryogenesis. In summary, a feasible in vivo assay for neurite outgrowth was established in zebrafish. PMID- 15358205 TI - Acute effect of corticosterone on N-methyl-D-aspartate receptor-mediated Ca2+ elevation in mouse hippocampal slices. AB - We examined the rapid effects of corticosterone (CORT) on N-methyl-D-aspartate (NMDA) receptor-mediated Ca2+ signals in adult mouse hippocampal slices by using Ca2+ imaging technique. Application of NMDA caused a transient elevation of intracellular Ca2+ concentration followed by a decay to a plateau within 150s. The 30min preincubation of CORT induced a significant decrease of the peak amplitude of NMDA-induced Ca2+ elevation in the CA1 region. The rapid effect of CORT was induced at a stress-induced level (0.4-10microM). Because the membrane non-permeable bovine serum albumin-conjugated CORT also induced a similar rapid effect, the rapid effect of CORT might be induced via putative surface CORT receptors. In contrast, CORT induced no significant effects on NMDA-induced Ca2+ elevation in the dentate gyrus. In the CA3 region, CORT effects were not evaluated, because the marked elevation of NMDA-induced Ca2+ signals was not observed there. PMID- 15358206 TI - Transcription, translation, degradation, and circadian clock. AB - Synthesis and degradation of mRNA together with synthesis and degradation of corresponding protein, this four-step-expression confers great fitness to all organisms. Transcription rate and mRNA stability both are essential for circadian expression of clock genes. In many cases, transcription rates and half-lives of mRNAs and corresponding proteins are not necessarily tightly linked with each other. The methods for measuring four-step-expression should be carefully selected and the experimental conditions should be strictly controlled. PMID- 15358207 TI - GM-CSF cannot substitute for M-CSF in human osteoclastogenesis. AB - Osteopetrotic mice lacking functional M-CSF recover with ageing, suggesting alternate osteoclastogenesis pathways exist. One alternative is GM-CSF, treatment with which improves the osteopetrosis. Our objective was to determine whether GM CSF could replace M-CSF in human osteoclastogenesis in vitro. Human CFU-GM precursors cultured with RANKL differentiate into osteoclasts without added M CSF, indicating constitutive production of M-CSF. Addition of M-CSF antibody completely inhibited differentiation, demonstrating M-CSF-dependence in vitro. Co treatment with low concentrations (0.01 ng/mL) of GM-CSF for 14 days or higher concentrations (10 ng/mL) for the first 1-2 days enhanced osteoclastogenesis but this effect was blocked with M-CSF antibody. Treatment with GM-CSF transiently increased M-CSF mRNA expression at 3 h but suppressed expression at 7-14 days. Neither FLT3-ligand nor VEGF supported osteoclastogenesis in the absence of M CSF. Thus, in vitro human osteoclastogenesis is dependent on M-CSF and the stimulatory effects of GM-CSF are mediated by M-CSF. Rescue by GM-CSF in M-CSF deficiency is unlikely to be directly mediated by FLT3-ligand or VEGF. PMID- 15358208 TI - Synergistic activation of the CMV promoter by NF-kappaB P50 and PKG. AB - Several DNA binding NF-kappaB subunits are substrates for cGMP-dependent kinase (PKG) and their transactivation from cognate sites is induced by phosphorylation. This includes p50, which does not have a transcriptional activation domain and therefore needs to bind to other proteins to mediate gene expression. Here, we describe the synergistic transactivation by p50 and PKG from the CMV promoter. This is caused not only by phosphorylation of p50, leading to increased DNA binding, but also by PKG-dependent activation of CRE sites in the promoter. One of the CRE sites is located directly adjacent to a NF-kappaB site and is essential for p50-mediated induction of transcription. According to the binding of CREB to p50 in pull-down assays and according to the inhibition of p50 dependent transactivation by dominant-negative CREB, this reflects the formation of a transcription factor complex containing CREB and p50. The nuclear translocation of NF-kappaB is insufficient to distinguish among the multitude of promoters that harbor cognate recognition sites. The phosphorylation of multiple transcription factors by an upstream kinase, such as PKG, can lead to the formation of transcription factor complexes and differential transactivation from a subset of NF-kappaB sites. These interactions may be relevant for the activation of viral gene expression. PMID- 15358209 TI - Cadmium inhibits human DNA mismatch repair in vivo. AB - The heavy metal cadmium (Cd) is a human carcinogen that inhibits DNA repair activities. We show that DNA mismatch repair (MMR)-mediated cell cycle arrest after alkylation damage is suppressed by exposure to Cd and that this effect is reversed by preincubation with excess of zinc (Zn). We show that Cd-mediated inactivation of MMR activity is not caused by disruption of complex formation between the MMR proteins hEXO1-hMutS alpha and hEXO1-hMutL alpha nor does Cd inhibit 5'-exonuclease activity of hEXO1 in vitro. Thus, our studies show that exposure of human cells to Cd suppresses MMR activity, a repair activity known to play an important role in colon cancer and that this effect can be reversed by Zn treatment. PMID- 15358210 TI - A novel phosphoprotein is induced during bone marrow commitment to dendritic cells. AB - Dendritic cells (DCs) play an important role in vertebrate immunity, but little is known of the molecular events associated with their development from bone marrow (BM). This report describes induction of a signature protein marking BM commitment to DCs. Using a standard procedure, DCs were generated from BM by cultivation in vitro. Appropriate phenotypic monitoring was done primarily by immunofluorescence, and polyclonal antibody reagents were developed against immature DC lysates. Using one specific antibody reagent, we identified, purified, and sequenced a unique cytosolic phosphoprotein DP58 that occurs within 30 min during BM commitment to DCs. Its sequence matches with a computationally predicted Riken cDNA (GenBank Accession No. XP_138799), and a specific anti-DP58 peptide antibody was developed for further characterization. The study suggests that DP58 induction signals distinct pathway(s) leading to early DC progenitors that may be generated and propagated for a short period in vitro. PMID- 15358211 TI - Chaperone characteristics of PDI-related protein A from Aspergillus niger. AB - The functional properties of a novel protein, protein disulfide isomerase-related protein A (PRPA) from Aspergillus niger T21, have been characterized. (1) PRPA possesses disulfide isomerase activity. (2) In Hepes buffer, at substoichiometric concentrations, PRPA facilitates the formation of inactive lysozyme aggregates associated with PRPA (anti-chaperone activity); while at a high molar excess, PRPA inhibits aggregation by maintaining lysozyme in a soluble, yet inactive, state (chaperone-like activity). However, PRPA only exhibits chaperone-like activity during lysozyme refolding in phosphate buffer. (3) Experiments have indicated that disulfide cross-linkage is not required for the interaction between PRPA and lysozyme, and hydrophobic interaction may be responsible for PRPA effect on lysozyme. (4) Co-expression of PRPA and prochymosin in Escherichia coli leads to reduction of inclusion bodies, rendering part of prochymosin molecules soluble yet inactive. The structural and functional characteristics of PRPA suggest that PRPA may play an important role in protein folding, aggregation, and retention in the endoplasmic reticulum. PMID- 15358212 TI - Skewed X-inactivation in cloned mice. AB - In female mammals, dosage compensation for X-linked genes is accomplished by inactivation of one of two X chromosomes. The X-inactivation ratio (a percentage of the cells with inactivated maternal X chromosomes in the whole cells) is skewed as a consequence of various genetic mutations, and has been observed in a number of X-linked disorders. We previously reported that phenotypically normal full-term cloned mouse fetuses had loci with inappropriate DNA methylation. Thus, cloned mice are excellent models to study abnormal epigenetic events in mammalian development. In the present study, we analyzed X-inactivation ratios in adult female cloned mice (B6C3F1). Kidneys of eight naturally produced controls and 11 cloned mice were analyzed. Although variations in X-inactivation ratio among the mice were observed in both groups, the distributions were significantly different (Ansary-Bradley test, P<0.01). In particular, 2 of 11 cloned mice showed skewed X inactivation ratios (19.2% and 86.8%). Similarly, in intestine, 1 of 10 cloned mice had a skewed ratio (75.7%). Skewed X-inactivation was observed to various degrees in different tissues of different individuals, suggesting that skewed X inactivation in cloned mice is the result of secondary cell selection in combination with stochastic distortion of primary choice. The present study is the first demonstration that skewed X-inactivation occurs in cloned animals. This finding is important for understanding both nuclear transfer technology and etiology of X-linked disorders. PMID- 15358213 TI - Alkaline serine protease produced by Streptomyces sp. degrades PrP(Sc). AB - A PrP(Sc)-degrading enzyme was isolated from the culture medium of Streptomyces sp. using perchloric acid-soluble protein (PSP) as a substrate. The media of 500 microbial species were screened to obtain the PSP-degrading enzyme. The medium containing the protease secreted from strain 99-GP-2D-5 showed the highest PSP degrading activity. Strain 99-GP-2D-5 was assigned as the genus Streptomyces by its morphological and chemotaxonomic characteristics. When scrapie prion was used as the substrate, it was completely digested by the enzyme. The amino acid sequence of the enzyme was identical to that of the C-terminal region of alkaline serine protease (ASP) I. ASP I may be the precursor of the enzyme, and the enzyme seems to be the mature type of ASP I. The maximal activity of the enzyme was observed at 60 degrees C and pH 11, and the scrapie prion was degraded within 3 min under the optimum conditions. PMID- 15358214 TI - Homeostatic regulation of the proteasome via an Rpn4-dependent feedback circuit. AB - The 26S proteasome is a complex protease consisting of at least 32 different subunits. Early studies showed that Rpn4 (also named Son1 and Ufd5) is a transcriptional activator of the Saccharomyces cerevisiae proteasome genes, and that Rpn4 is rapidly degraded by the 26S proteasome. These observations suggested that in vivo proteasome abundance may be regulated by an Rpn4-dependent feedback circuit. Here, we present direct evidence to support the Rpn4-proteasome feedback model. We show that proteasome expression is increased when proteasome activity is impaired, and that this increase is Rpn4-dependent. Moreover, we demonstrate that expression of a stable form of Rpn4 leads to elevation of proteasome expression. Our data also reveal that the Rpn4-proteasome feedback circuit is critical for cell growth when proteasome activity is compromised, and plays an important role in response to DNA damage. This study provides important insights into the mechanism underlying proteasome homeostasis. PMID- 15358215 TI - Runx3-/- gastric epithelial cells differentiate into intestinal type cells. AB - We have previously reported that Runx3, a runt domain transcription factor, is a major growth regulator of gastric epithelial cells, that a lack of RUNX3 function is causally related to the genesis and progression of human gastric cancer, and that expression of RUNX3 is greatly reduced in intestinal metaplasias in human stomachs. Here we examined the differentiation of Runx3-/- mouse gastric epithelial cells and found that some cells differentiated into intestinal type cells, which expressed Cdx2, a transcription factor that has been shown to induce intestinal metaplasia in transgenic mice. Differentiation of intestinal type cells was not found in culture of Runx3+/+ gastric epithelial cells. These results suggest that gastric epithelial cells can differentiate into intestinal type cells, probably due to expression of Cdx2 in them when the function of Runx3 is impaired. The relationship between loss of function of Runx3, formation of intestinal metaplasia, and gastric cancer was discussed. PMID- 15358216 TI - C-reactive protein impairs angiogenic functions and decreases the secretion of arteriogenic chemo-cytokines in human endothelial progenitor cells. AB - C-reactive protein (CRP), a predictor of future cardiovascular diseases, has been reported to damage the vascular wall by inducing endothelial dysfunction and inflammation. This proatherogenic CRP was speculated to have a role in attenuating angiogenic functions of human endothelial progenitor cells (EPCs), possibly impairing vascular regeneration and increasing cardiovascular vulnerability to ischemic injury. Herein, we investigated the direct effect of CRP on angiogenic activity and gene expression in human EPCs. Incubation of EPCs with human recombinant CRP significantly inhibited EPC migration in response to vascular endothelial growth factor, possibly by decreasing the expression of endothelial nitric oxide synthase and subsequent nitric oxide production. In addition, CRP-treated EPCs showed the reduced adhesiveness onto an endothelial cell monolayer. When assayed for the gene expression of arteriogenic chemo cytokines, CRP substantially decreased their expression levels in EPC, in part due to the upregulation of suppressors of cytokine signaling proteins. These results suggest that CRP directly attenuates the angiogenic and possibly arteriogenic functions of EPCs. This CRP-induced EPC dysfunction may impair the vascular regenerative capacity of EPCs, thereby leading to increased risk of cardiovascular diseases. PMID- 15358217 TI - Identification of polymorphisms in the promoter region of the human NRF2 gene. AB - Transcription factor Nrf2 regulates the basal and inducible expression of detoxifying and antioxidant genes. Recent studies using nrf2-null mice suggest that Nrf2 dysfunction might be involved in the pathogenesis of human diseases. To gain insight into the relationship between impairment in the NRF2 gene and human diseases, we attempted to identify polymorphisms in the human NRF2 gene. We determined the structure of the NRF2 gene and found three single nucleotide polymorphisms and one triplet repeat polymorphism in its regulatory region. These results provide a molecular basis for the genetic analysis of the NRF2 gene. The frequency of each polymorphism was examined in two groups of patients with systemic lupus erythematosus and chronic obstructive pulmonary disease. This study did not reveal a close connection between the risk of these diseases and the polymorphisms. However, available lines of evidence suggest the importance of examining the link between NRF2 polymorphisms and other oxidative stress-related diseases. PMID- 15358218 TI - Cadmium induces the expression of specific stress proteins in sea urchin embryos. AB - Marine organisms are highly sensitive to many environmental stresses, and consequently, the analysis of their bio-molecular responses to different stress agents is very important for the understanding of putative repair mechanisms. Sea urchin embryos represent a simple though significant model system to test how specific stress can simultaneously affect development and protein expression. Here, we used Paracentrotus lividus sea urchin embryos to study the effects of time-dependent continuous exposure to subacute/sublethal cadmium concentrations. We found that, between 15 and 24 h of exposure, the synthesis of a specific set of stress proteins (90, 72-70, 56, 28, and 25 kDa) was induced, with an increase in the rate of synthesis of 72-70 kDa (hsps), 56 kDa (hsp), and 25 kDa, which was dependent on the lengths of treatment. Recovery experiments in which cadmium was removed showed that while stress proteins continued to be synthesized, embryo development was resumed only after short lengths of exposure. PMID- 15358219 TI - Ion channel activity of membrane vesicles released from sea urchin sperm during the acrosome reaction. AB - The sperm acrosome reaction (AR) involves ion channel activation. In sea urchin sperm, the AR requires Ca2+ and Na+ influx and K+ and H+ efflux. During the AR, the plasma membrane fuses with the acrosomal vesicle membrane forming hybrid membrane vesicles that are released from sperm into the medium. This paper reports the isolation and preliminary characterization of these acrosome reaction vesicles (ARVs), using synaptosome-associated protein of 25 kDa (SNAP-25) as a marker. Isolated ARVs have a unique protein composition. The exocytosis regulatory proteins vesicle-associated membrane protein and SNAP-25 are inside ARVs, as judged by protease protection experiments, and membrane associated based on Triton X-114 partitioning. ARVs fused with planar bilayers display three main types of single channel activity. The most frequently recorded channel is cationic, weakly voltage dependent and has a low open probability that increases with negative potentials. This channel is activated by cAMP, blocked by Ba2+, and has a PK+/PNa+ selectivity of 4.5. ARVs represent a novel membrane preparation suitable to deepen our understanding of ion channel activity in the AR and during fertilization. PMID- 15358220 TI - Expression of selenocysteine-containing glutathione S-transferase in Escherichia coli. AB - Evolution of a probable 'glutathione-binding ancestor' resulting in a common thioredoxin-fold for glutathione S-transferases and glutathione peroxidases may possibly suggest that a glutathione S-transferase could be engineered into a selenium-containing glutathione S-transferase (seleno-GST), having glutathione peroxidase (GPX) activity. Here, we addressed this question by production of such protein. In order to obtain a recombinant seleno-GST produced in Escherichia coli, we introduced a variant bacterial-type selenocysteine insertion sequence (SECIS) element which afforded substitution with selenocysteine for the catalytic Tyr residue in the active site of GST from Schistosoma japonica. Utilizing coexpression with the bacterial selA, selB, and selC genes (encoding selenocysteine synthase, SelB, and tRNA(Sec), respectively) the yield of recombinant seleno-GST was about 2.9 mg/L bacterial culture, concomitant with formation of approximately 85% truncation product as a result of termination of translation at the selenocysteine-encoding UGA codon. The mutations inferred as a result of the introduction of a SECIS element did not affect the glutathione binding capacity (Km = 53 microM for glutathione as compared to 63 microM for the wild-type enzyme) nor the GST activity (kcat = 14.3 s(-1) vs. 16.6 s(-1)), provided that the catalytic Tyr residue was intact. When this residue was changed to selenocysteine, however, the resulting seleno-GST lost the GST activity. It also failed to display any novel GPX activity towards three standard peroxide substrates (hydrogen peroxide, butyl hydroperoxide or cumene hydroperoxide). These results show that recombinant selenoproteins with internal selenocysteine residues may be heterologously produced in E. coli at sufficient amounts for purification. We also conclude that introduction of a selenocysteine residue into the catalytic site of a glutathione S-transferase is not sufficient to induce GPX activity in spite of a maintained glutathione-binding capacity. PMID- 15358221 TI - In vitro mesengenic potential of human umbilical cord blood-derived mesenchymal stem cells. AB - Human mesenchymal stem cells (hMSCs) have been paid a great deal of attention because of their unprecedented therapeutic merits endowed by powerful ex vivo expansion and multilineage differentiation potential. Umbilical cord blood (UCB) is a convenient but not fully proven source for hMSCs, and hence, greater experience is required to establish UCB as a reliable source of hMSCs. To this end, we attempted to isolate hMSC-like adherent cells from human UCB. The isolated cells were highly proliferative and exhibited an immunophenotype of CD13+ CD14- CD29+ CD31- CD34- CD44+ CD45- CD49e+ CD54+ CD90+ CD106- ASMA+ SH2+ SH3+ HLA-ABC+ HLA-DR-. More importantly, these cells, under appropriate conditions, could differentiate into a variety of mesenchymal lineage cells such as osteoblasts, chondrocytes, adipocytes, and skeletal myoblasts. This mesengenic potential assures that the UCB-derived cells are multipotent hMSCs and further implicates that UCB can be a legitimate source of hMSCs. PMID- 15358222 TI - Antibacterial synergism of novel antibiotic peptides with chloramphenicol. AB - HP (2-20) is an antimicrobial sequence derived from the N-terminus of Helicobacter pylori ribosomal protein L1. We previously tested whether several analogues of HP (2-20), with amino acid substitutions that increased or decreased net hydrophobicity, could be useful as therapeutic agents. In the present study, we show that substituting Gln and Asp for Trp at positions 17 and 19, respectively, of HP (2-20) (peptide A3) had potent antibacterial activity in minimal inhibition concentration and minimal bactericidal concentration without having hemolytic activity. In contrast, when we decreased hydrophobicity by substituting Leu or Phe for Ser at positions 12 and 19, respectively, of HP (2 20) (Anal 4, Anal 5), there was no significant effect on antibacterial activity. We found that A3 acted synergistically with chloramphenicol against bacterial cells. Fluorescence activated flow cytometry showed that A3-treated cells had higher fluorescence intensity than untreated cells, similar to that of melittin treated cells. Furthermore, A3 caused significant morphological alterations of Staphylococcus aureus and Pseudomonas aeruginosa, as shown by scanning electron microscopy. Our results suggest that peptide A3 may be useful for the design of novel antibiotic peptides that possess high bacterial cell selectively and synergistic effects with conventional antibiotic agents but lack hemolytic activity. PMID- 15358223 TI - Inhibitory effect of n-3 fish oil fatty acids on cardiac Na+/Ca2+ exchange currents in HEK293t cells. AB - Abnormal activity of the cardiac Na+/Ca2+ exchanger (NCX1) can affect intracellular Ca2+ homeostasis and cause arrhythmias. The n-3 polyunsaturated fatty acids (PUFAs), however, may prevent arrhythmias. To test the effect of PUFAs on the cardiac NCX1 current (I(NCX1)), the canine NCX1 cDNA was expressed in human embryonic kidney (HEK293t) cells. The average density of I(NCX1) was 10.9+/-2.6 pA/pF (n=44) in NCX1-transfected cells and eicosapentaenoic acid (EPA, C20:5n-3) significantly inhibited I(NCX1) The suppression of I(NCX1) by EPA was concentration-dependent with an IC50 of 0.82+/-0.27 microM. EPA had a similar effect on outward or inward I(NCX1). Docosahexaenoic acid (DHA, C22:6n-3) and arachidonic acid (AA, C20:4n-6) also significantly inhibited I(NCX1), whereas the saturated fatty acid, stearic acid (SA, C18:0), did not. Our data demonstrate that the n-3 PUFAs significantly suppress cardiac I(NCX1), which is probably one of their protective effects against lethal arrhythmias. PMID- 15358224 TI - Differential induction of gene promoter constructs by constitutively active human TLRs. AB - Antigen presenting cells can sense microorganisms through activation of members of the Toll like receptor family (TLRs), which initiate signals leading to transcription of many inflammation-associated genes. TLRs and IL-1R, through their TIR domains, activate NFkappaB and mitogen-activated protein kinase pathways and upregulate a set of specific target genes. Recent evidence points to several differences in signaling pathways activated by individual TLRs. To evaluate the basic signaling potential of individual TIR signaling domains, we generated constitutively active versions of all known human TLRs by fusing mouse CD4 extracellular portion with the TLR transmembrane and TIR domains. A panel of promoters from genes known to be activated by TLRs as well as artificial promoter constructs with transcription factor binding sites were selected to measure their response in the presence of constitutively active CD4TLR fusion molecules. These studies show for the first time that a unique panel of promoters appears to be highly induced by CD4TLR1, 6 (TLRs that usually function through heterodimerisation with TLR2), and CD4TLR10. We also observed that CD4TLR4 is the most potent gene activator compared to all other ten human TLRs. Preliminary analyses of several promoter deletions showed that TLRs use different sequence elements to activate these reporters. In addition, since different ligands for a single TLR (e.g., TLR9) can induce different pathways, the CD4TLR fusions seem to activate all the pathways and therefore can be used to assess the overall signaling capacity of a given TLR. Finally, analysis of promoter constructs induced by the only orphan TLR, TLR10, allowed the identification of the ENA78 promoter as a tool for screening its ligands. PMID- 15358225 TI - Induction of heme oxygenase-1 is involved in anti-proliferative effects of paclitaxel on rat vascular smooth muscle cells. AB - In this study, we evaluated the possibility that the anti-proliferative effects of paclitaxel on vascular smooth muscle cells (VSMCs) of the rat might be due to the induction of HO-1 gene expression. Treatment of the cells with paclitaxel resulted in marked time- and dose-dependent inductions of HO-1 mRNA, followed by corresponding increases in HO-1 protein expression and HO enzymatic activities. Furthermore, paclitaxel rapidly activated the JNK, ERK, and p38 mitogen-activated protein kinase pathways. A specific inhibitor of JNK, SP600125, abolished paclitaxel-induced HO-1 mRNA expression, whereas PD98059, a specific inhibitor of ERK, and SB203580, a specific inhibitor of p38, had no significant effect. Finally, the suppression of platelet-derived growth factor induced VSMC proliferation was abolished by the HO inhibitor, ZnPP, as well as by the CO scavenger, hemoglobin. These results demonstrated that paclitaxel induces the expression of HO-1 via the JNK pathway in VSMC and that HO-1 expression might be responsible for the anti-proliferative effect of paclitaxel on VSMC. PMID- 15358226 TI - The malaria parasite mitochondrion senses cytosolic Ca2+ fluctuations. AB - By using the fluorescent dye Rhod-2, we have investigated the ability of Plasmodium mitochondria to participate in cellular Ca2+ homeostasis. To this end, isolated parasites were simultaneously loaded with the mitochondrial Ca2+ probe Rhod-2 and the cytosolic Ca2+ dye Fluo-3 and their fluorescent intensities were monitored in the same cells by confocal microscopy. We here demonstrate that Ca2+ increases, as elicited by treatment of parasites with sarco-endoplasmic reticulum Ca2+ ATPase inhibitors or the hormone melatonin, induce rapid and reversible increases of the Ca2+ concentration in the mitochondria of both human and murine parasites. Pre-treatment of parasites with the mitochondrial uncoupler, FCCP, suppresses mitochondrial Ca2+ accumulation. Our data demonstrate that mitochondria of malaria parasites are able to reversibly accumulate part of the Ca2+ released in the cytoplasm by pharmacological and physiological agents and thus suggest that this organelle participate in the maintenance of Ca2+ homeostasis of Plasmodia. PMID- 15358227 TI - Transcriptional regulation of SLP-76 family hematopoietic cell adaptor MIST/Clnk by STAT5. AB - SLP-76-related adaptor protein MIST (also called Clnk) is expressed in a variety of cytokine-dependent hematopoietic cell lines of myeloid and lymphoid origin as well as some cytokine-independent mast cell lines. To understand the molecular mechanisms underlying the MIST gene expression, we have characterized the 5' flanking region of the mouse MIST gene. We have identified an enhancer region ( 773 to -709), which is active in P815 mast cells expressing the endogenous MIST gene, but not in EL-4 T cells lacking MIST expression. Outside of this enhancer region, one STAT element present in the MIST promoter (-44 to -36) was found to bind STAT5A when IC-2 mast cells were stimulated with IL-3. Mutation of this STAT element did not affect basal MIST promoter activity in P815 mast cells, but was required for STAT5-mediated activation of the MIST promoter. Furthermore, endogenous MIST gene expression was induced in mast cells by a constitutively activated form of STAT5A, but not by an active mutant of c-Kit receptor. These findings suggest that STAT5 is involved in cytokine-mediated up-regulation of MIST gene expression, probably in collaboration with other lineage-specific transcription factors that promote basal MIST expression in mast cells. PMID- 15358228 TI - Dual action of adiponectin on insulin secretion in insulin-resistant mice. AB - Adiponectin is secreted by adipocytes and has been implicated as a mediator of insulin sensitivity. In this study, the acute effects of adiponectin on islets isolated from normal or diet-induced insulin resistant mice were examined. In normal islets, adiponectin (5 microg/ml) had no significant effect on insulin secretion. In contrast, in islets from mice rendered insulin resistant by high fat feeding, adiponectin inhibited insulin secretion at 2.8 mM (P < 0.01) but augmented insulin secretion at 16.7 mM glucose (P < 0.05). The augmentation of glucose-stimulated insulin secretion by adiponectin was accompanied by increased glucose oxidation (P < 0.005), but without any significant effect on palmitate oxidation or the islet ATP/ADP ratio. Furthermore, RT-PCR revealed the expression of the adiponectin receptor AdipoR1 mRNA in mouse islets, however, with no difference in the degree of expression level between the two feeding groups. The results thus uncover a potential dual role for adiponectin to modify insulin secretion in insulin resistance. PMID- 15358229 TI - AMP-activated protein kinase activators can inhibit the growth of prostate cancer cells by multiple mechanisms. AB - Prostate cancer cells require high rates of de novo fatty acid synthesis and protein synthesis for their rapid growth. We report here that the growth of these cells is markedly diminished by incubation with activators of AMP-activated protein kinase (AMPK), a fuel-sensing enzyme that has been shown to diminish both of these processes in intact tissues. Inhibition of cell growth was observed when AMPK was activated by either 5-aminoimidazole-4-carboxamide riboside (AICAR) or the thiazolidinedione rosiglitazone. Thus, a 90% inhibition of the growth of androgen-independent (DU145, PC3) and androgen-sensitive (LNCaP) cells was achieved after 4 days of exposure to one or both of these agents. Where studied, this was associated with a decrease in the concentration of malonyl CoA, an intermediate of de novo fatty acid synthesis, and an increase in expression of the cell cycle inhibitor p21. In addition, AICAR inhibited two key enzymes involved in protein synthesis, mTOR and p70S6K, and blocked the ability of the androgen R1881 to increase cell growth and the expression of two enzymes for de novo fatty acid synthesis, acetyl CoA carboxylase and fatty acid synthase, in the LNCaP cells. The results suggest that AMPK is a potential target for the treatment of prostate cancer. PMID- 15358230 TI - Transplantation of human umbilical cord blood cells improves glycemia and glomerular hypertrophy in type 2 diabetic mice. AB - Recent in vitro and in vivo studies have shown that either animal- or human derived embryonic stem cells can differentiate into insulin-secreting cells and lower blood glucose levels. However, studies utilizing human umbilical cord blood (HUCB) mononuclear cells to improve blood glucose levels in diabetic animals have received little attention. In this study, we examined the effect of transplanted HUCB mononuclear cells on blood glucose levels, survival, and renal pathology in obese mice with spontaneous development of type 2 diabetes. The results show that injection of HUCB mononuclear cells into orbital plexus of mice caused improvement not only in blood glucose levels and survival rate but also normalization of glomerular hypertrophy and tubular dilatation. Thus, transplantation of HUCB mononuclear cells appears to be another modality of stem cell therapy in diabetes mellitus. PMID- 15358231 TI - Identification of the minimal repression domain of SUPERMAN shows that the DLELRL hexapeptide is both necessary and sufficient for repression of transcription in Arabidopsis. AB - We reported previously that the carboxy-terminal 30 amino acids of SUPERMAN (SUPRD) function as a repression domain in Arabidopsis. In this study, we identified the peptide sequences in SUPRD that is both necessary and sufficient for repression of transcription. To our surprise, the hexapeptide DLELRL was sufficient, by itself, to confer the ability to repress transcription on a DNA binding domain. A database search revealed that there are 32 TFIIIA-type zinc finger proteins in the Arabidopsis genome that contain a hexapeptide sequence similar or identical to that of DLELRL. These peptides acted as repression domains, suggesting that these zinc finger proteins might function as active repressors. Further mutational analysis within DLELRL revealed that an amphiphilic motif composed of six amino acids (XLxLXL) with preferences at the first and fifth positions is necessary and sufficient for strong repression. An assay of positional effects suggested that GAL4DB-DLELRL might function as a short-range repressor. A possible mechanism of the DLELRL-mediated repression is discussed. PMID- 15358232 TI - Exercise normalises overexpression of TNF-alpha in knockout mice. AB - TNF-alpha is linked with insulin resistance, as greater amounts of TNF are detected in muscle and adipose tissue in glycemically challenged people and TNF alpha inhibits insulin receptor signalling. However, what modulates this overexpression of TNF-alpha is currently unknown. We examined the effect of 1 h exercise on overexpression of the TNF-alpha gene in TNF receptor 1 and 2 knockout mice. IL-6 knockout mice were included to elucidate the importance of IL-6 in regulating TNF-alpha in response to exercise. TNF-alpha gene expression was over expressed in muscle in both TNFR knockout models. TNF-alpha overexpression returned to normal levels after exercise in the TNF-alpha receptor knockout models. In IL-6 knockout mice, a modest decrease in TNF-alpha was also observed. These data suggest that TNF-alpha-induced insulin resistance can be regulated by a single exercise bout by normalising TNF-alpha expression. This exercise effect can be mediated via IL-6, but also an IL-6 independent mechanism seems to exist. PMID- 15358233 TI - Human NTH1 physically interacts with p53 and proliferating cell nuclear antigen. AB - Thymine glycol (Tg) is one of predominant oxidative DNA lesions caused by ionizing radiation and other oxidative stresses. Human NTH1 is a bifunctional enzyme with DNA glycosylase and AP lyase activities and removes Tg as the first step of base excision repair (BER). We have searched for the factors interacting with NTH1 by using a pull-down assay and found that GST-NTH1 fusion protein precipitates proliferating cell nuclear antigen (PCNA) and p53 as well as XPG from human cell-free extracts. GST-NTH1 also bound to recombinant FLAG-tagged XPG, PCNA, and (His)6-tagged p53 proteins, indicating direct protein-protein interaction between those proteins. Furthermore, His-p53 and FLAG-XPG, but not PCNA, stimulated the Tg DNA glycosylase/AP lyase activity of GST-NTH1 or NTH1. These results provide an insight into the positive regulation of BER reaction and also suggest a possible linkage between BER of Tg and other cellular mechanisms. PMID- 15358234 TI - Expression and characterization of cholera toxin B-pneumococcal surface adhesin A fusion protein in Escherichia coli: ability of CTB-PsaA to induce humoral immune response in mice. AB - Cholera toxin B subunit (CTB) is responsible for CT holotoxin binding to the cell and has been described as a mucosal adjuvant for vaccines. In this work, the ctxB gene was genetically fused to the psaA gene from Streptococcus pneumoniae, a surface protein involved in its colonization in the host that is also considered a vaccine antigen candidate against this pathogen. The CTB-PsaA fusion protein was expressed in Escherichia coli, and the purified protein was used for intranasal immunization experiments in Balb/C mice. CTB-PsaA was able to induce both systemic and mucosal antibodies evaluated in serum, saliva, and in nasal and bronchial wash samples, showing that CTB-PsaA is a promising molecule to be investigated as S. pneumoniae vaccine antigen candidate. PMID- 15358235 TI - Plasmodium falciparum expresses a multidrug resistance-associated protein. AB - Plasmodium falciparum proteins that efflux toxic metabolic products such as oxidised glutathione (GSSG) are possible targets for anti-malarial drug development. Proteins capable of transporting GSSG and glutathione conjugates include the multidrug resistance-associated transporters (MRPs). A gene, PFA0590w, encoding a MRP homologue, has been identified in P. falciparum. Here we show the presence of full-length mRNA (5.5 kb) of this PfMRP in trophozoites by RT-PCR and Northern blotting. A polyclonal anti-PfMRP antibody generated against two unique, hydrophilic peptides in the predicted sequence produced a strong immunoreactive protein band of 210-215 kDa on Western blots of schizonts of chloroquine-sensitive and chloroquine-resistant strains, confirming expression of PfMRP protein. Using confocal microscopy the protein was seen to be localised at the edge of the schizonts with no obvious staining of the food vacuole. We suggest that PfMRP may act as the GSSG transporter in the parasite plasma membrane. PMID- 15358236 TI - A novel soluble protein factor with non-opioid dynorphin A-binding activity. AB - A novel soluble non-opioid dynorphin A-binding factor (DABF) was identified and characterized in neuronal cell lines, rat spinal cord, and brain. DABF binds dynorphin A(1-17), dynorphin A(2-17), and the 32 amino acid prodynorphin fragment big dynorphin consisting of dynorphin A and B, but not other opioid and non opioid peptides, opiates, and benzomorphans. The IC50 for dynorphin A(1-17), dynorphin A(2-17), and big dynorphin is in the 5-10 nM range. Using dynorphin A and big dynorphin fragments a binding epitope was mapped to dynorphin A(6-13). DABF has a molecular mass of about 70 kDa. SH-groups are apparently involved in the binding of dynorphin A since p-hydroxy-mercuribenzoic acid inhibited this process. Upon interaction with DABF dynorphin A was converted into Leu enkephalin, which remained bound to the protein. These data suggest that DABF functions as an oligopeptidase that forms stable and specific complexes with dynorphin A. The presence of DABF in brain structures and other tissues with low level of prodynorphin expression suggests that DABF as an oligopeptidase may degrade other peptides. Dynorphin A at the sites of its release in the CNS may attenuate this degradation as a competitor when it specifically binds to the enzyme. PMID- 15358237 TI - Identification of novel phosphorylation sites on postsynaptic density proteins. AB - Phosphorylation of the components of the postsynaptic density (PSD), a protein complex lining the postsynaptic membrane, may regulate synaptic structure and function. We carried out mass spectrometric analyses to identify phosphorylation sites on PSD proteins. Phosphopeptides were isolated from the total tryptic digest of a PSD fraction by immobilized metal affinity chromatography and analyzed by liquid chromatography and tandem mass spectrometry. The phosphorylated residues detected following in vitro phosphorylation in the presence of Ca2+/calmodulin included S-1058 on SynGAP and S-1662 and S-1668 on Shank3. Other phosphorylated residues were identified in control samples, presumably reflecting phosphorylation in the intact cell. These included the homologous residues, S-295 on PSD-95 and S-365 on PSD-93, located between the PDZ2 and PDZ3 domains of these proteins; and S-367 located on the actin-binding domain of beta-CaMKII. The sequence RXXSPV emerged as a common phosphorylation motif of three specialized PSD scaffolding proteins, PSD-95, PSD-93, and Shank3. Phosphorylated serine residues in several of the identified phosphorylation sites were followed by prolines, suggesting prominent involvement of proline directed kinases in the regulation of PSD components. PMID- 15358238 TI - Transient receptor potential vanilloid subfamily 1 expressed in pancreatic islet beta cells modulates insulin secretion in rats. AB - Capsaicin-sensitive afferent neurons including transient receptor potential vanilloid subfamily 1, TRPV1, and neurohormonal peptides participate in the physiological regulation of pancreatic endocrine. However, the direct effect of capsaicin on insulin secretion remains unknown. Our present study showed that TRPV1 is expressed in islet beta cells as well as in neurons in rat pancreas, and also in rat beta cell lines, RIN and INS1. Capsaicin (10(-11)-10(-9) M) dose dependently increased insulin secretion from RIN cells, and this effect was inhibited by either a TRPV1 inhibitor capsazepine or EDTA. Systemic capsaicin (10 mg/kg, s.c.) increased plasma insulin level 1 h after the treatment. We demonstrated for the first time that TRPV1 is functionally expressed in rat islet beta cells and plays a role in insulin secretion as a calcium channel. This study may account for the influences of capsaicin on the food intake and energy consumption as well as on the pathophysiological regulation of pancreatic endocrine. PMID- 15358239 TI - Ligand-induced alpha2-adrenoceptor endocytosis: relationship to Gi protein activation. AB - Most G protein-coupled receptors are desensitized by a uniform two-step mechanism: phosphorylation followed by arrestin binding and internalization. In this study we explored the time-, ligand-, and concentration dependence of alpha2 adrenoceptor internalization in human embryonal kidney (HEK-293) cells expressing alpha2A- and alpha2B-adrenoceptors. We also explored the relationship between ligand-induced receptor internalization and agonist efficacy, determined with a [35S]GTPgammaS binding assay. The results showed rapid dose-dependent internalization of both alpha2A- and alpha2B-receptors; the extent of internalization was directly proportional to agonist efficacy. The agonist UK 14,304 had a subtype-specific high efficacy at alpha2A-AR and dexmedetomidine at alpha2B-AR. Agonist-induced [35S]GTPgammaS binding was totally blocked by pretreatment with pertussis toxin (PTX) for both receptor subtypes, while only about 50% of the internalization was blocked by PTX. The results indicate that the extent of internalization of alpha2A-AR and alpha2B-AR is proportional to agonist efficacy, but only partly dependent on Gi protein coupling. PMID- 15358240 TI - Insights into function of PSI domains from structure of the Met receptor PSI domain. AB - PSI domains are cysteine-rich modules found in extracellular fragments of hundreds of signaling proteins, including plexins, semaphorins, integrins, and attractins. Here, we report the solution structure of the PSI domain from the human Met receptor, a receptor tyrosine kinase critical for proliferation, motility, and differentiation. The structure represents a cysteine knot with short regions of secondary structure including a three-stranded antiparallel beta sheet and two alpha-helices. All eight cysteines are involved in disulfide bonds with the pattern consistent with that for the PSI domain from Sema4D. Comparison with the Sema4D structure identifies a structurally conserved core comprising the N-terminal half of the PSI domain. Interestingly, this part links adjacent SEMA and immunoglobulin domains in the Sema4D structure, suggesting that the PSI domain serves as a wedge between propeller and immunoglobulin domains and is responsible for the correct positioning of the ligand-binding site of the receptor. PMID- 15358241 TI - Carbon monoxide exposure in rat heart: evidence for two modes of toxicity. AB - Rat hearts were perfused for 30 min with buffer equilibrated with CO. Mean data for hearts exposed to 0.01% CO show a 15% decrease in heart rate (HR) during exposure followed by recovery, with a further 17% decrease post exposure. Examination of time courses from individual perfusions shows that in 10 hearts exposed to 0.01% CO HR responded in different ways: no response (5 hearts); decrease during exposure followed by recovery (3 hearts); and decrease post exposure (2 hearts). There was a strong association between CO-induced HR decrease and release of creatine kinase into the perfusate, both of which were not prevented by the antioxidants, ascorbate, and Trolox C. Perfusate flow rate declined post exposure (4.9% and 8.9% with 0.01% and 0.05% CO, respectively) and this was prevented by antioxidants. CO may have two, independent, cardiotoxic effects; these may be mediated by CO-induced elevation of oxidant production, H2O2 in one case and peroxynitrite in the other. PMID- 15358242 TI - A comparison of siRNA efficacy predictors. AB - Short interfering RNA (siRNA) efficacy prediction algorithms aim to increase the probability of selecting target sites that are applicable for gene silencing by RNA interference. Many algorithms have been published recently, and they base their predictions on such different features as duplex stability, sequence characteristics, mRNA secondary structure, and target site uniqueness. We compare the performance of the algorithms on a collection of publicly available siRNAs. First, we show that our regularized genetic programming algorithm GPboost appears to have a higher and more stable performance than other algorithms on the collected datasets. Second, several algorithms gave close to random classification on unseen data, and only GPboost and three other algorithms have a reasonably high and stable performance on all parts of the dataset. Third, the results indicate that the siRNAs' sequence is sufficient input to siRNA efficacy algorithms, and that other features that have been suggested to be important may be indirectly captured by the sequence. PMID- 15358243 TI - AlphaB-crystallin inhibits glucose-induced apoptosis in vascular endothelial cells. AB - Recent studies implicate hyperglycemia as a cause of vascular complications in diabetes. Our study confirmed that high concentration of glucose (30 mM) induces apoptosis in cultures of human umbilical vein endothelial cells. After 5 days of culture TUNEL positive cells in high concentration of glucose were nearly 63% higher when compared to normal concentration of glucose (5 mM). Transfection of pcDNA3-rat alphaB-crystallin into these cells inhibited high glucose-induced apoptosis by approximately 36%, such an effect was not observed when cells were transfected with an empty vector. AlphaB-crystallin transfection inhibited by about 35% of high glucose induced activation of caspase-3. High concentration of glucose enhanced formation of reactive oxygen species (ROS) in these cells but this was significantly (p < 0.001) curtailed by transfection of alphaB crystallin. Results of our study indicate that alphaB-crystallin effectively inhibits both ROS formation and apoptosis in cultured vascular endothelial cells and provide a basis for future therapeutic interventions in diabetic vascular complications. PMID- 15358244 TI - Cloning of hOST-PTP: the only example of a protein-tyrosine-phosphatase the function of which has been lost between rodent and human. AB - Protein-tyrosine-phosphatases (PTP-ases), in concert with protein tyrosine kinases, control various biological activities such as cell growth and differentiation. In rodents, around 40 PTP-ases have been described. Functional orthologue for each of these PTP-ases have been identified in human, except for OST-PTP. OST-PTP is a transmembrane PTP-ase with a restricted tissue distribution. In silico analysis on public sequence databases reveals a human OST PTP gene orthologue that encompasses 21 kb on chromosome 1q32.1. Using RT-PCR we isolated a 4 kb hOST-PTP transcript. hOST-PTP cDNA sequence exhibits numerous disablements indicating that it does not code for a PTP-ase but is rather a pseudogene with unique features. Indeed, (i) it has no "functional" parent in the human genome, (ii) it has retained an "intron-exon" structure, and (iii) it is transcribed in a regulated manner. Interestingly, we found two ESTs, from domesticated pig and from cow that exhibit ORF that would predict a functional OST-PTP orthologue in Artiodactyls. Taken together, these results indicate that OST-PTP is the only PTP-ase the function of which has been lost during the evolution process between rodents and human. PMID- 15358246 TI - Meeting the information needs of people with cancer. PMID- 15358251 TI - Proteomics on its way to study host-pathogen interaction in Candida albicans. AB - Candida albicans is a common fungal pathogen of humans, but also exists as a commensal in the population. Proteomics of C. albicans has been used since the early 1980s, however, only the recent publication of the genome sequence of C. albicans and improvements in mass spectrometry technologies have made it possible to apply proteomics to C. albicans on a larger scale. This includes analysing the cell wall, investigating drug response or changes in mutants with defects in virulence. In addition, serological responses to systemic candidiasis have been monitored and screens for virulence factors using patient sera, have been described. These promising approaches are just emerging, anticipating further contributions in C. albicans proteomics that will advance our understanding of host-pathogen interaction in the near future. PMID- 15358252 TI - Candida albicans cell wall glycans, host receptors and responses: elements for a decisive crosstalk. AB - Candida albicans has adapted to live on the mucosal surfaces of animals. The human species has accepted it. By contrast to numerous other commensals, C. albicans has a prominent ability to invade virtually all tissues of a host presenting with natural or acquired defects in homeostasis. C. albicans uses considerable energy to synthesize glycans, which are present either as polymers or as glyconjugates. These glycan molecules play a prominent role in the biology of C. albicans by controlling the structure and plasticity of the cell wall, and are also involved in yeast-host interactions. These glycans are recognized as 'non-self' by host innate and adaptative immunity. The signal they induce in the host depends on the 'glycan code', which is determined by the nature of the sugar, the anomer type of linkage and branching, and the length of the oligosaccharide chains. However, this model is not static because the nature of the C. albicans molecule carrying such glycan codes and their expression at the cell wall surface also determines the host response, and, in turn, the regulation of cell wall glycan arrangement dynamics in C. albicans depends on host stimuli. Candida glycans therefore play an important role in the continuous interchange that regulates the balance between saprophytism and parasitism, and resistance and infection. A goal of current research concerning the virulence attributes of C. albicans will be to determine to what extent this species is able to regulate its glycan code as a response to the host. PMID- 15358253 TI - Candida morphogenesis and host-pathogen interactions. AB - The human fungal pathogen Candida albicans has many morphological forms. Recent advances in genomics and cell biology are providing an improved understanding of the molecular regulation of cell shape, and providing insights into the relationships between morphogenesis and virulence. This understanding may improve our ability to develop strategies to combat Candida infections. PMID- 15358255 TI - Cysteine peptidases as virulence factors of Leishmania. AB - Leishmania mexicana amastigotes are particularly rich in cysteine peptidases (CPs), which play important roles in facilitating the survival and growth of the parasites in mammals. The importance of the CPs as virulence factors and their potential as drug targets and vaccine candidates has been investigated extensively. Recent years, however, have heralded advances in our knowledge and understanding of leishmanial CPs on two fronts. Firstly, genome analysis has revealed the great diversity of CPs, and, secondly, the ways in which the most widely studied CPs, designated CPB, influence the interaction between parasite and mammalian host have been elucidated. These topics are the focus of this review. PMID- 15358256 TI - Pernicious plans revealed: Plasmodium falciparum genome wide expression analysis. AB - The asexual intraerythrocytic developmental cycle (IDC) of Plasmodium falciparum is responsible for the majority of the clinical manifestations of malaria in humans. Although malaria has been studied for over a century, the elucidation of the full genome sequence of P. falciparum has now allowed for in-depth studies of gene expression throughout the entire intraerythrocytic stage. As the mainstays of anti-malarial chemotherapy become increasingly ineffective, we need a deeper understanding of fundamental plasmodial bioregulatory mechanisms to successfully subvert them. Recent gene expression studies have begun to examine different aspects of the IDC and are providing key insights into the basic mechanisms of Plasmodium gene regulation and are helping to define gene functions. However, to date, no transcription factor has been fully characterized from Plasmodium and the definitive identification of cis-acting regulatory elements along with their corresponding trans-acting partners is still lacking. The characterization of the transcriptome of P. falciparum is the first major step towards the understanding of the genome wide regulation of gene expression in this parasite. IDC expression data for almost every gene in the P. falciparum genome can now be publicly queried at and. The results of these studies suggest promising leads for identifying novel targets for anti-malarial therapeutics and vaccines in addition to providing a solid foundation for the ongoing elucidation of plasmodial gene expression. PMID- 15358257 TI - Host cell invasion by the apicomplexans: the significance of microneme protein proteolysis. AB - Intracellular life-style has been adopted by many pathogens as a successful immune evasion mechanism. To gain entry to a large variety of host cells and to establish an intracellular niche, Toxoplasma gondii and other apicomplexans rely on an active process distinct from phagocytosis. Calcium-regulated secretion of microneme proteins and parasite actin polymerization together with the action of at least one myosin motor act in concert to generate the gliding motility necessary to propel the parasite into host cells. During this active penetration, host cell transmembrane proteins are excluded from the forming parasitophorous vacuole hence conferring the resistance to acidification and degradative fusion. Apicomplexans possess a large repertoire of microneme proteins that contribute to invasion, but their precise role and the level of functional redundancy remain to be evaluated. Remarkably, most microneme proteins are proteolytically cleaved during biogenesis and post-exocytosis. The significance of the processing events and the identification of the proteases implicated are the object of intensive investigations. These proteases may constitute potential drug targets for intervention against malaria and other diseases caused by these parasites. PMID- 15358258 TI - Host-microbe interactions: viruses a never-ending creativity contest. PMID- 15358259 TI - Foamy viruses--a world apart. AB - Foamy viruses (FVs) or spumaviruses were described for the first time in the early 1950s in cell cultures derived from monkey kidneys. Later, FVs were isolated in several mammal species such as cats, cattle and horses. Highly prevalent in non-human primates they are not naturally present in humans, although several cases of simian-to-human transmissions have been described. Interestingly, the replication strategy of FVs differs in many aspects from that of other retroviruses, presenting features that are closely related to pararetroviruses, exemplified by the hepatitis B virus (HBV), but also characteristics that are closely related to yeast retrotransposons. These characteristics led to the creation of a distinct viral subfamily by the International Committee on Virus Taxonomy in 2002; the Spumaretrovirinae. PMID- 15358260 TI - HIV-host interactions: vital to the virus and key to its inhibition. AB - Much progress has been made in recent years in the investigation of the interplay between HIV-1 and its host cells. Most of these interactions are complex and have not yet been fully unraveled. Nevertheless, current knowledge on the molecular interactions between HIV and host-cell factors has substantially broadened our understanding of the viral life cycle and opened new investigative areas for drug intervention. PMID- 15358261 TI - Molecular biology of severe acute respiratory syndrome coronavirus. AB - The worldwide epidemic of severe acute respiratory syndrome (SARS) in 2003 was caused by a novel coronavirus called SARS-CoV. Coronaviruses and their closest relatives possess extremely large plus-strand RNA genomes and employ unique mechanisms and enzymes in RNA synthesis that separate them from all other RNA viruses. The SARS epidemic prompted a variety of studies on multiple aspects of the coronavirus replication cycle, yielding both rapid identification of the entry mechanisms of SARS-CoV into host cells and valuable structural and functional information on SARS-CoV proteins. These recent advances in coronavirus research have important implications for the development of anti-SARS drugs and vaccines. PMID- 15358262 TI - Psychotropic viruses. AB - Viruses are increasingly being recognized as important factors in the pathogenesis of acute and chronic mental illness. Here we review clinical and epidemiologic data concerning viral infection and mental illness, as well as animal models that provide insight into the myriad of mechanisms by which infection can cause brain dysfunction. PMID- 15358263 TI - Public, private and non-specific antibodies induced by non-cytopathic viral infections. AB - Lymphocytic choriomeningitis virus (LCMV) represents a useful experimental model of murine infection with a non-cytopathic virus, bearing resemblance to HIV and hepatitis C virus (HCV) infections in humans. Recent data from the LCMV model indicate that the humoral immune response that is induced by non-cytopathic viruses is far more complex than previously appreciated. LCMV-induced IgG production is largely polyclonal, with more than 90% of the antibody repertoire constituting non-relevant specificities. A delayed virus-neutralizing antibody response is induced, including specificities directed not only against the parental LCMV-strain present in the host but also cross-specifically against LCMV variants isolated from other hosts. These findings provide novel insights to aid our understanding of clinically relevant observations that are recorded following human infection with HIV, HCV and dengue viruses. PMID- 15358265 TI - From Arabidopsis to agriculture: engineering improved Vitamin E content in soybean. AB - For more than 80 years, tocopherols have been known to be an essential nutrient, vitamin E, for humans and animals. Work in recent years has concentrated on dissecting tocopherol biosynthesis to engineer the pathway in agricultural crops. Molecular dissection of the pathway in plants is now complete with the cloning and characterization of the gene for Arabidopsis MPBQ/MSBQ methyltransferase (VITAMIN E 3; VTE3). Alison Van Eennemaan and colleagues used seed-specific expression of two vitamin E pathway methyltransferases to engineer increased vitamin E activity in soybeans. PMID- 15358266 TI - Structure of biological solar energy converters - further revelations. AB - Photosynthetic organisms harvest solar energy by absorbing light and ultimately transferring energy through a cascade of chemical reactions to power all cellular processes. Core components initiating this reaction cascade are the photosynthetic reaction centres Photosystem I and Photosystem II. Two recent publications on the structure of the reaction centres by Adam Ben-Shem et al. and Kristina Ferreira et al. represent a big step towards understanding the evolutionary development of the core energy conversion process and identifying the site of the water oxidation process, the source of atmospheric oxygen. PMID- 15358267 TI - The ALDH gene superfamily of Arabidopsis. AB - Aldehyde dehydrogenases (ALDHs) represent a protein superfamily of NAD(P)(+) dependent enzymes that oxidize a wide range of endogenous and exogenous aliphatic and aromatic aldehydes. The Arabidopsis genome contains 14 unique ALDH sequences encoding members of nine ALDH families, including eight known families and one novel family (ALDH22) that is currently known only in plants. Here, we identify members of the ALDH gene superfamily in Arabidopsis; provide a revised, unified nomenclature for these ALDH genes; analyze the molecular relationship among Arabidopsis ALDH genes and compare them to ALDH genes from other species, including prokaryotes and mammals; and describe the role of ALDHs in cytoplasmic male sterility, plant defense and abiotic stress tolerance. PMID- 15358268 TI - Learning the lipid language of plant signalling. AB - Plant cells respond to different biotic and abiotic stresses by producing various uncommon phospholipids that are believed to play key roles in cell signalling. We can predict how they work because animal and yeast proteins have been shown to have specific lipid-binding domains, which act as docking sites. When such proteins are recruited to the membrane locations where these phospholipids are synthesized, the phospholipids activate them directly, by inducing a conformational change, or indirectly, by juxtaposing them with an activator protein. The same lipid-binding domains are present in Arabidopsis proteins. We believe that they represent an untapped well of information about plant lipid signalling. PMID- 15358269 TI - Insights into the molecular dynamics of plant light-harvesting proteins in vivo. AB - To understand physiological processes at the molecular level, new techniques are needed to determine the details of protein structure and dynamics in intact systems. We describe a specific example of such an approach, involving differential analysis of the carotenoid resonance Raman signal in the plant photosynthetic membrane. Carotenoids play important roles in the photosynthetic membrane and are particularly vital to photoprotective regulatory mechanisms. Our methodology selectively revealed the details of associations between specific carotenoid molecules and specific protein binding sites. Changes in the molecular configuration of these cofactors associated with alterations in the physiological state of the photosynthetic system were observed. This approach can be applied to a wide range of complex biological systems, whenever a protein with a light absorbing cofactor is involved. PMID- 15358270 TI - Weed genomics: new tools to understand weed biology. AB - In spite of the large yield losses that weeds inflict on crops, we know little about the genomics of economically important weed species. Comparative genomics between plant model species and weeds, map-based approaches, genomic sequencing and functional genomics can play vital roles in understanding and dissecting weedy traits of agronomically important weed species that damage crops. Weed genomics research should increase our understanding of the evolution of herbicide resistance and of the basic genetics underlying traits that make weeds a successful group of plants. Here, we propose specific weed candidates as genomic models, including economically important plants that have evolved herbicide resistance on several occasions and weeds with good comparative genomic qualities that can be anchored to the genomics of Arabidopsis and Oryza sativa. PMID- 15358271 TI - Antifreeze proteins in overwintering plants: a tale of two activities. AB - Antifreeze proteins are found in a wide range of overwintering plants where they inhibit the growth and recrystallization of ice that forms in intercellular spaces. Unlike antifreeze proteins found in fish and insects, plant antifreeze proteins have multiple, hydrophilic ice-binding domains. Surprisingly, antifreeze proteins from plants are homologous to pathogenesis-related proteins and also provide protection against psychrophilic pathogens. In winter rye (Secale cereale), antifreeze proteins accumulate in response to cold, short daylength, dehydration and ethylene, but not pathogens. Transferring single genes encoding antifreeze proteins to freezing-sensitive plants lowered their freezing temperatures by approximately 1 degrees C. Genes encoding dual-function plant antifreeze proteins are excellent models for use in evolutionary studies to determine how genes acquire new expression patterns and how proteins acquire new activities. PMID- 15358272 TI - A primer on molecular biology for imagers: VI. Proteomics: the large-scale study of proteins. PMID- 15358274 TI - Matrix metalloproteinase-dependent EGF receptor activation in hypertension and left ventricular hypertrophy. AB - Agonist stimulation of certain G protein-coupled receptors (GPCRs) causes shedding of heparin-binding epidermal growth factor (HB-EGF) through activation of matrix metalloproteinases (MMPs), with subsequent transactivation of the EGF receptor. MMPs are widely expressed, and their dysregulated expression is crucial in cancer, inflammation, and cardiovascular remodeling. Recent studies in hypertensive animals have shown enhanced expression and activation of MMPs and EGF receptors, and their inhibition attenuates cardiac hypertrophy, vasoconstriction and hypertension induced by GPCR agonists such as angiotensin II, endothelin-1 and phenylepherine. These findings suggest that selective inhibition of MMPs might have therapeutic potential in hypertension and other cardiovascular diseases. PMID- 15358275 TI - Flying through the genome: a comprehensive study of functional genomics using RNAi in Drosophila. AB - Sequencing the DNA of an entire mammalian genome now seems routine. The human sequence along with the mouse--the model for mammalian genetics--and the rat--the model for mammalian physiology--are now part of the data archive. However, the real challenges for the 21st century are what to do with this information and how to test the function of so many different genes in so many different cellular contexts. The potential payoffs are enormous. Examples include a better understanding of disease pathologies with effective strategies for therapeutic interventions that cause few, if any, side effects. PMID- 15358276 TI - Resistin/ADSF/FIZZ3 in obesity and diabetes. AB - The role of adipocyte-secreted resistin/adipocyte-specific secretory factor (ADSF)/FIZZ3 in obesity and diabetes has been controversial at best. Recently generated resn knockout mice showed normal glucose and insulin sensitivity with lower fasting glucose levels. Upon feeding with a high-fat diet, the knockout mice exhibited increased glucose tolerance with decreased hepatic glucose output, possibly due to phosphorylation and activation of AMP-activated protein kinase and suppression of gluconeogenic genes. In comparison, transgenic mice overexpressing a dominant negative form of resistin/ADSF/FIZZ3 showed increased adiposity with elevated leptin and adiponectin levels, accompanying enhanced glucose tolerance and insulin sensitivity both on chow and high-fat diets. Although its underlying mechanisms need further elucidation, the in vivo studies demonstrate a role of resistin/ADSF/FIZZ3 in obesity and insulin resistance. PMID- 15358277 TI - Alternatives to growth hormone stimulation testing in children. AB - Despite more than 40 years of pediatric growth hormone (GH) replacement, we are still limited in our ability to make a definitive diagnosis of GH deficiency (GHD) in children. Historically, GH stimulation tests (GHSTs) have been used to discriminate between GHD and idiopathic short stature. Over the years, increases in the peak diagnostic GH cutoffs and the proliferation of GH assays have fundamentally changed the nature of the GHST. In our opinion, today's GHSTs lack reproducibility and accuracy, are expensive, and can be dangerous. Moreover, newer diagnostic tools, such as high-resolution neuroimaging, measurements of serum insulin-like growth factor 1 and insulin-like growth factor-binding protein 3, and an increasing number of genetic tests, have emerged. We believe that it is no longer appropriate to use GHSTs to diagnose childhood GHD. Instead, diagnosis should be based on a combination of auxological, biochemical, neuroradiological and genetic considerations. Here, we examine the alternatives to the GHST that are currently available and literature that supports their use. We believe that these alternative methods should replace the GHST. PMID- 15358278 TI - Gut hormones and the control of appetite. AB - Obesity is the main cause of premature death in the UK. Worldwide its prevalence is accelerating. It has been hypothesized that a gut nutriment sensor signals to appetite centres in the brain to reduce food intake after meals. Gut hormones have been identified as an important mechanism for this. Ghrelin stimulates, and glucagon like peptide-1, oxyntomodulin, peptide YY (PYY), cholecystokinin and pancreatic polypeptide inhibit, appetite. At physiological postprandial concentrations they can alter food intake markedly in humans and rodents. In addition, in obese humans fasting levels of PYY are suppressed and postprandial release is reduced. Administration of gut hormones might provide a novel and physiological approach in anti-obesity therapy. Here, we summarize some of the recent advances in this field. PMID- 15358279 TI - Mineralocorticoid resistance. AB - Mineralocorticoid resistance, also known as type I pseudohypoaldosteronism (PHA1), is a rare inherited disease characterized by salt wasting, dehydration and failure to thrive in the newborn. Two different forms of the disease, which present with either systemic or exclusively renal resistance to aldosterone, are associated with two different modes of inheritance. The generalized, recessive form of the disease is due to abnormalities in the epithelial sodium channel, and inactivating mutations of the mineralocorticoid receptor are responsible for the autosomal dominant form of PHA1 and some sporadic cases. Here, we review the role of aldosterone in the maintenance of normal sodium balance and discuss the different forms of mineralocorticoid resistance and the underlying genetic abnormalities. Although important progress has been made in the past years, there remain several families in whom the genetic defect has not been identified. Precise clinical diagnosis and establishment of intermediate phenotypes should be helpful for identifying other genes involved in PHA1 and gaining new insight into the regulation of sodium homeostasis. PMID- 15358280 TI - Imaging evaluation of the non-functioning indeterminate adrenal mass. AB - With the increasing use of abdominal cross-sectional imaging in the investigation of patient symptoms and in cancer staging, incidental adrenal masses are frequently detected. The most common clinical question is whether these masses are benign or malignant. Benign adrenal masses such as myelolipomas, lipid-rich adenomas, adrenal cysts and adrenal haemorrhage have pathognomonic imaging findings. However, there remains a significant overlap between the imaging appearances of some lipid-poor adenomas and malignant lesions, particularly metastases and small adrenal carcinomas. Our review looks at the recent advances in computed tomography, magnetic resonance imaging and positron emission tomography, which can be used to assist in the distinction between benign adenomas and malignant lesions of the adrenal gland. PMID- 15358281 TI - Progestogen therapies: differences in clinical effects? AB - A large number of estrogen/progestogen preparations are available for the treatment of estrogen-deficiency symptoms. These preparations differ in the route of administration, the type and dose of both the estrogen and progestogen. The only indication for the addition of a progestogen is endometrial protection, but, depending on its chemical structure, a progestogen can either enhance (e.g. hot flushes, gonadotropin release, breast-epithelial proliferation and bone mineral density) or antagonize (e.g. endometrium, arterial wall, lipid metabolism, hepatic protein synthesis and mood) the effects of the estrogen component. Available progestogens differ largely in their hormonal pattern and, in addition to their progestogenic and antiestrogenic action on the endometrium, they can exert androgenic, antiandrogenic, glucocorticoid and/or antimineralocorticoid effects. There are no comprehensive trials comparing directly the modulating effects of the various progestogens, and clinical and epidemiological data do not allow a definite conclusion on the clinical relevance of differences between progestogens. PMID- 15358282 TI - Intracellular trafficking of hormone receptors. AB - Agonist binding stimulates endocytosis of hormone receptors via vesicular uptake mechanisms. Interactions of the intracellular domains of receptors with specific targeting proteins are crucial for sorting of internalized receptor in endosomes. Some receptors are targeted for very rapid (e.g. beta2-adrenergic receptor) or slower (e.g. AT1 angiotensin receptor) recycling pathways, whereas others are targeted to lysosomes for degradation (e.g. EGF receptor or PAR1 protease activated receptor). This review discusses the mechanisms involved in these processes, which regulate surface receptor expression and set the stage for intracellular signaling of G protein-coupled and growth factor receptors. PMID- 15358283 TI - Novel tetrahydro-thieno pyridyl oxazolidinone: an antibacterial agent. AB - Synthesis of a number of 4,5,6,7-tetrahydro-thieno[3,2-c]pyridine substituted oxazolidinones have been reported. They have been screened against a panel of Gram-positive pathogens including methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis. A SAR has been developed. Compound 15 showed comparable activity (MIC) to linezolid and superior to eperezolid. PMID- 15358284 TI - Protein phosphatase 2A inhibition and circumvention of cisplatin cross-resistance by novel TCM-platinum anticancer agents containing demethylcantharidin. AB - Novel TCM-platinum compounds [Pt(C(8)H(8)O(5))(NH(2)R)(2)] 1-5, derived from integrating demethylcantharidin, a modified component from a traditional Chinese medicine (TCM) with a platinum moiety, possess anticancer and protein phosphatase 2A inhibition properties. The compounds are able to circumvent cisplatin resistance by apparently targeting the DNA repair mechanism. Novel isosteric analogues [Pt(C(9)H(10)O(4))(NH(2)R)(2)] A and B, devoid of PP2A-inhibitory activity, were found to suffer from an enhanced DNA repair and were cross resistant to cisplatin. The results advocate a well-defined structure-activity requirement associating the PP2A-inhibiting demethylcantharidin with the circumvention of cisplatin cross-resistance demonstrated by TCM-Pt compounds 1-5. PMID- 15358285 TI - Tumor promoter binding of the protein kinase C C1 homology domain peptides of RasGRPs, chimaerins, and Unc13s. AB - Recent investigations discovered nonkinase-type phorbol ester receptors, RasGRPs, chimaerins, and Unc13s. Phorbol ester binding occurs at the cysteine-rich sequences of about 50 residues in the C1 domains of these receptors. Fifty-one residue RasGRP C1 peptides except for RasGRP2 showed significant phorbol 12,13 dibutyrate (PDBu) binding, but the K(d) values of the RasGRP1 and RasGRP3 C1 peptides were about 10-fold larger than those for the corresponding whole enzymes. Addition of the C-terminal basic amino acid cluster decreased their K(d) values about 10-fold, suggesting that the positive charges of these C1 peptides play an important role in the PDBu binding in the presence of negatively-charged phosphatidylserine. The 51-mer chimaerin C1 peptides showed potent PDBu binding, while the Unc13 and Munc13-1 C1 peptides without sufficient positive charges hardly bound PDBu. By the rapid screening system using this C1 peptide library, 5 prenyl-indolactam-V was identified as a promising lead for the novel protein kinase C isozyme specific ligands. PMID- 15358286 TI - Non-thiol farnesyltransferase inhibitors: N-(4-tolylacetylamino-3-benzoylphenyl) 3-arylfurylacrylic acid amides. AB - We have designed arylfurylacryl-substituted benzophenones as non-thiol farnesyltransferase inhibitors utilizing a novel aryl binding site of farnesyltransferase. These compounds display activity in the low nanomolar range. PMID- 15358287 TI - Chronobiotic activity of N-[2-(2,7-dimethoxyfluoren-9-yl)ethyl]-propanamide. Synthesis and melatonergic pharmacology of fluoren-9-ylethyl amides. AB - A series of fluoren-9-yl ethyl amides (2) were synthesized and evaluated for human melatonin MT(1) and MT(2) receptor binding. N-[2-(2,7-dimethoxyfluoren-9 yl)ethyl]propanamide (2b) was selected and evaluated in functional assays measuring intrinsic activity at the human MT(1) and MT(2) receptors and demonstrated full agonism at both receptors. The chronobiotic properties of 2b were demonstrated in both acute and chronic rat models where 2b produced an acute phase advance of 32 min at 1mg/kg and chronically entrained free-running rats with a mean effective dose of 0.23 mg/kg. Compound 2b is significantly less efficacious than melatonin in constricting human coronary artery. PMID- 15358288 TI - Synthesis, acute toxicities, and antitumor effects of novel 9-substituted beta carboline derivatives. AB - A series of novel 9-substituted beta-carboline derivatives was synthesized from harmine and l-tryptophan, respectively. Cytotoxic activities of these compounds in vitro were investigated. The results showed that most compounds of 9 substituted beta-carboline derivatives had more remarkable cytotoxic activities in vitro than their corresponding parent compounds. Acute toxicities and antitumor effects of the selected beta-carboline derivatives in mice were also examined. The results demonstrated that a short alkyl or benzyl substituent at position-9 increased the antitumor activities significantly and a ethoxycarbonyl or carboxyl substituent at position-3 reduced the acute toxicity and neurotoxicity of these beta-carboline derivatives dramatically. Moreover the compounds both with an alkoxycarbonyl or carboxyl substituent at position-3 and a short alkyl or benzyl substituent at positon-9 exhibited more significant antitumor activities and lower acute toxicities and neurotoxicities than the other compounds. The compound 8c, having an n-butyl and a carboxyl substituent at position-9 and 3, respectively, was found to have the highest antitumor effect and the lowest acute toxicity and neurotoxicity. These data suggested that (1) appropriate substituents at both position-9 and 3 of beta-carboline derivatives might play a crucial role in determining their enhanced antitumor activities and decreased acute toxicities and neurotoxic effects; (2) the beta-carboline derivatives have the potential to be used as antitumor drug leads. PMID- 15358289 TI - Analogues of tetramethylrosamine as transport molecules for and inhibitors of P glycoprotein-mediated multidrug resistance. AB - Tetramethylrosamine and its thio- and seleno- analogues (TMR-O, TMR-S, and TMR Se, respectively) were examined for their ability to be transported by Pgp into chemo-resistant CR1R12 cells. Verapamil (7 x 10(-6)M) enhanced the uptake of TMR O and TMR-S into CR1R12 cells compared to those cultures not previously exposed to verapamil. The uptake of TMR-O and TMR-S in CR1R12 cells in the presence of 7 x 10(-6)M verapamil was equivalent to its uptake in the chemo-sensitive parent cell line AUXB1 in the absence or presence of verapamil. None of the TMR analogues were effective alone as photosensitizers of CR1R12 cells. However, when either TMR-S or TMR-Se was added to CR1R12 cells after 7 x 10(-6)M verapamil exposure for 2h, irradiation of cultures with 5.0J cm(-2) of 350-750 nm light caused significant phototoxicity. TMR-O showed no significant phototoxicity in the presence of verapamil. Chemo-sensitive AUXB1 cells are equally susceptible to phototoxicity using TMR-Se with or without previous exposure to verapamil. The Pgp modulators verapamil and CsA increased the uptake of CAM into CR1R12. Exposure of CR1R12 cells to TMR-S or TMR-Se for 2h in the dark resulted in no significant change in the intracellular accumulation of CAM. However, 1h of light exposure after incubation of cells with TMR-S or TMR-Se resulted in an up to 2 fold increase in CAM uptake. PMID- 15358290 TI - Syntheses and evaluation of antioxidant activity of sydnonyl substituted thiazolidinone and thiazoline derivatives. AB - 3-Aryl-4-formylsydnone 4'-phenylthiosemicarbazones (3a-d) and 3-aryl-4 formylsydnone thiosemicarbazones (3e-h), which are precursors of 3-aryl-4 heterocyclic sydnones, are prepared by the condensation of 3-aryl-4 formylsydnones (1a-d) with 4'-phenylthiosemicarbazide (2a) and thiosemicarbazide (2b), respectively. The thiosemicarbazones 3 reacted with cyclic reagents such as ethyl chloroacetate (4a), ethyl 2-chloroacetoacetate (4b) and 2-bromoacetophenone (4c) to produce heterocyclic substituted sydnone derivatives 5-7 that possess 4 oxo-thiazolidine and thiazoline groups. The antioxidant activity of synthesized compounds 5a-7h was evaluated. Among these compounds, 4-methyl-2-[(3-arylsydnon-4 yl-methylene)hydrazono]-2,3-dihydro-thiazole-5-carboxylic acid ethyl ester (6e-h) and 4-phenyl-2-[(3-arylsydnon-4-yl-methylene)hydrazono]-2,3-dihydro-thiazoles (7e h) exhibit the potent DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, comparable to that of vitamin E. PMID- 15358291 TI - Highly potent PDE4 inhibitors with therapeutic potential. AB - The hypothesis that the dose-limiting side effects of PDE4 inhibitors could be mediated via the central nervous system prompted us to design and synthesize a hydrophilic piperidine analog to improve the side effect profile of Ariflo 1, which is an orally active second-generation PDE4 inhibitor. During evaluation of various water-soluble piperidine analogs, 2a-b, 11b-14b, and 17a showed therapeutic potential in cross-species comparison studies. The following three findings were obtained: (1) The hydroxamic acid group, a well known metal chelator, caused a marked increase of inhibitory activity. (2) Water-soluble piperidine analogs lacked the configurational isomerism of Ariflo 1 without loss of inhibitory activity. (3) Replacement of the 4-methoxy residue with a difluoromethoxy residue led to an increase of in vivo potency. Structure-activity relationships are presented. Single-dose rat pharmacokinetic data for 11b, 12b, and 17a are also presented. PMID- 15358292 TI - Design, synthesis, and pharmacological evaluation of some 2-[4-morpholino]-3-aryl 5-substituted thiophenes as novel anti-inflammatory agents: generation of a novel anti-inflammatory pharmacophore. AB - Compounds incorporating a thiophene moiety, a pi excess five membered heterocycle, have attracted a great deal of research interest owing to the therapeutic utility of the template as useful drug molecular scaffolding. Recently we reported the anti-inflammatory activity profile exhibited by two thiophene analogs, AP84 and AP82 in acute and chronic models of inflammation. The good activity profile exhibited by AP84, a 3-(substituted aryl)-2-(4-morpholino) 5-heteroaryl substituted analog of thiophene, in the formalin induced rat paw edema chronic model as compared to a weak activity in acute carrageenin induced rat paw edema, and the slightly better protection exhibited in the acute model by AP82 (27%), the 5-aroyl analog provided an impetus for a proper exploration of their structural types. In this paper we report the synthesis and pharmacological evaluation of some novel, 2-(4-morpholino)-3-(substituted aryl)-5-substituted thiophenes, as possible anti-inflammatory leads. The 3-(4-chlorophenyl)-2-(4 morpholino) thiophene analogs AP49, AP158, and AP88 provided a protection of 20%, 23%, and 20%, respectively, when screened for anti-inflammatory activity in carrageenin induced rat paw edema, an acute in vivo model, comparable to that of AP82, at a dose level of 100mg/kg body weight p.o. compared to ibuprofen as standard. The replacement of the 3-(4-chlorophenyl) moiety with the 3-phenyl moiety gave rise to AP50 (30%), AP159 (38%), AP27 (0%), and AP92 (38%), with three analogs being more active in the acute model. Alteration of the group para to the phenyl ring at third position, from chloro, to methyl mercapto gave rise to the 3-(4-methylmercapto-phenyl) analogs AP54 (20%), AP160 (0%), and AP73 (52%), with only one analog appearing to be better than AP82. These results indicate that 4-methane sulfonyl aroyl group at 5-position and other substituents of different quadrants of Craig plot on the phenyl moiety at the third position could lead to more potent candidates. However, alteration of aroyl to substituted pyridyl at 5-position with a phenyl group at the third position as in AP26 gave rise to much better protection (66%) again reinforcing the importance of the heteroaryl ring at the fifth position and implying its utility in the composition of a novel pharmacophore for designing better trisubstituted thiophenes as anti inflammatory agents. PMID- 15358293 TI - Cytochrome P-450 dependent monooxygenases model system: rapid and efficient oxidation of primary aromatic amines to azo derivatives with sodium periodate catalyzed by manganese(III) Schiff base complexes. AB - Rapid and efficient oxidation of primary aromatic amines was investigated. Mn(III)-salophen catalyst can catalyze the oxidation of primary aromatic amines to azo derivatives with sodium periodate. The ability of various Schiff base complexes in this oxidation system was also investigated. PMID- 15358294 TI - Synthesis, characterization and in vitro anti-amoebic activity of new palladium(II) complexes with 5-nitrothiophene-2-carboxaldehyde N(4)-substituted thiosemicarbazones. AB - Reaction between [Pd(DMSO)(2)Cl(2)] (DMSO=dimethylsulfoxide) and N(4)-substituted thiosemicarbazones derived from 5-nitrothiophene-2-carboxaldehyde (L) afforded the complexes [Pd(L)Cl(2)]. These new complexes have been characterized by elemental analyses and spectroscopic studies. Spectroscopic studies reveal that thionic sulfur and azomethine nitrogen atom of thiosemicarbazones are coordinated to metal ion. The testing of the anti-amoebic activity of these complexes against the protozoan parasite Entamoeba histolytica suggests that compound 9, 10, and 11 might be endowed with important anti-amoebic properties since they showed less IC(50) values than metronidazole. Moreover, compound 11 displays notable amoebicidal activity than metronidazole (IC(50) values of 0.79 microM vs 1.93 microM, respectively). PMID- 15358295 TI - Development of prostaglandin D2 receptor antagonist: discovery of highly potent antagonists. AB - The process of discovery for highly potent prostaglandin D(2) (PGD(2)) receptor antagonists is reported. A series of N-(p-alkoxy)benzoyl-2-methylindole-4-acetic acids were synthesized and identified as a new class of selective PGD(2) receptor antagonists. Most of them exhibited strong PGD(2) receptor antagonism in binding studies and the cAMP formation assay. The structure-activity relationships (SAR), including subtype selectivity of the synthesized compounds, are also discussed. PMID- 15358296 TI - Design, synthesis, and biological evaluation of novel 4-alkylamino-1 hydroxymethylimidazo[1,2-a]quinoxalines as adenosine A(1) receptor antagonists. AB - A series of 4-alkylamino-1-hydroxymethylimidazo[1,2-a]quinoxalines have been synthesized and evaluated for their adenosine A(1) receptor inhibitory activity in the radioligand binding assays. The compounds were tested for the inhibition percent (IP) and the affinity toward A(1)AR (K(i)) that IP were more than 90% in the nanomolar range. 4-Cyclopentylamino-7,8-dichloro-1-hydroxymethylimidazo[1,2 a]quinoxaline 18 is the most potent compound in this series, having K(i)=7nM, which is remarkably higher than that of IRFI-165 (K(i)=48). 1-Hydroxymethyl groups of the tricyclic heteroarmatic compounds displayed the potent affinities toward A(1)AR. PMID- 15358297 TI - Antiradical and antioxidant activities of alkaloids isolated from Mahonia aquifolium. Structural aspects. AB - The antioxidant activities of three alkaloids isolated from Mahonia aquifolium- berberine, jatrorrhizine, and magnoflorine--were studied with respect to their structural aspects, particularly the presence and the position of -OH groups, steric conditions of unpaired electron delocalization and parameters of lipophilicity and hydration energy. The antiradical activities of the compounds tested were evaluated as the reactivities toward free stable alpha,alpha' diphenyl-beta-picrylhydrazyl radical (DPPH). The antioxidant features of the alkaloids tested were investigated in heterogeneous membrane system of DOPC liposomes stressed by peroxidative damage induced by AAPH azoinitiator. Both alkaloids bearing free phenolic groups--jatrorrhizine and magnoflorine--showed better activities in both systems used than berberine not bearing any readily abstractable hydrogen on its skeleton. The former two showed antiperoxidative efficiency in DOPC liposomal membrane comparable to that of an effective scavenger of peroxyl radicals--stobadine-and higher than that of Trolox. We conclude that the favorable antioxidant features of the hydroxylated alkaloids are most probably ensured by the combination of reasonably high antiradical reactivity with high lipophilicity, however, the solvation process was found to markedly interfere with these beneficial effects. PMID- 15358298 TI - Synthesis and evaluation of new hydrazide derivatives as neuropeptide Y Y5 receptor antagonists for the treatment of obesity. AB - NPY is the most potent orexigenic agent known to man, with NPY Y1 and NPY Y5 being the receptor subtypes that are most likely responsible for centrally mediated NPY-induced feeding responses. Based on the aforementioned, novel hydrazide derivatives were prepared for the purpose of searching new NPY Y5 receptor antagonists. Many of the compounds exhibited nanomolar binding affinity for this receptor, affording trans-N-(4-[N'-(3,4 dichlorophenyl)hydrazinocarbonyl]cyclohexylmethyl)-4-fluorobenzenesulfonamide, which showed the best activity (IC(50)=0.43nM). PMID- 15358299 TI - Induction of apoptosis in a leukemia cell line by triterpene saponins from Albizia adianthifolia. AB - Triterpenoid saponins, which are present in plants and some marine animals, exert various important pharmacological effects. The present study examines the effects of adianthifoliosides A, B, and D (AdA, AdB, and AdD) together with two prosapogenins (Pro1 and Pro2) obtained from Albizia adianthifolia (Mimosaceae) on human leukemia T-cells (Jurkat cells) and on splenocytes. AdA, AdB, and AdD were found to exhibit a cytotoxic effect on Jurkat cells, whereas the prosapogenins were found to exert a lymphoproliferative effect on this cell type. Furthermore, all tested compounds were found to exert a synergistic lymphoproliferative activity with concanavalin A (ConA) on splenocytes. The concentrations where the saponins were found to be cytotoxic on Jurkat cells are far below the concentration of hemolysis. These results indicate that another mechanism than membrane permeabilization formation is responsible of the cell cytotoxicity. Thus, we demonstrated that at 5 microM for AdA and at 1 microM for AdD, these compounds induce apoptosis in Jurkat cells. Early apoptotic events were detected by flow cytometry analysis by using a double annexin-V-FITC and propidium iodide staining. In addition, a disrupted mitochondrial membrane potential was observed in cells treated with AdA, AdB, and AdD. Furthermore, a DNA ladder was observed when Jurkat cells were incubated with 1 microM AdD for 24h. By comparison between the biological activities of the native compounds with the prosapogenins, we show in this work the important role of the acylation and esterification by different moieties at C-21 and C-28 of the aglycone (acacic acid) in the apoptosis-inducing capacity. Particularly, the monoterpene-quinovosyl moiety is shown to be important for the induction of apoptosis. PMID- 15358300 TI - Isoform-specific phosphoinositide 3-kinase inhibitors from an arylmorpholine scaffold. AB - Phosphoinositide 3-kinases (PI3-Ks) are an ubiquitous class of signaling enzymes that regulate diverse cellular processes including growth, differentiation, and motility. Physiological roles of PI3-Ks have traditionally been assigned using two pharmacological inhibitors, LY294002 and wortmannin. Although these compounds are broadly specific for the PI3-K family, they show little selectivity among family members, and the development of isoform-specific inhibitors of these enzymes has been long anticipated. Herein, we prepare compounds from two classes of arylmorpholine PI3-K inhibitors and characterize their specificity against a comprehensive panel of targets within the PI3-K family. We identify multiplex inhibitors that potently inhibit distinct subsets of PI3-K isoforms, including the first selective inhibitor of p110beta/p110delta (IC(50) p110beta=0.13 microM, p110delta=0.63 microM). We also identify trends that suggest certain PI3-K isoforms may be more sensitive to potent inhibition by arylmorpholines, thereby guiding future drug design based on this pharmacophore. PMID- 15358301 TI - Evaluation of DNA adduction of AZT in peripheral blood leukocytes of HIV-infected individuals by (32)P-post-labeling thin-layer chromatography: a feasibility study. AB - 3'-Azido-3'-deoxythymidine (AZT, Zidovudine) has been effectively used for HIV infection treatment. It inhibits virus reproduction through viral reverse transcriptase inhibition. However, the side effects of this anti-retroviral drug might be cumulative, particularly in its effects on the patients' DNA. As a nucleoside analogue, AZT might incorporate into hosts' DNA, and then form DNA adducts. This may result in potential long-term risks of mutagenesis in AIDS patients who received therapy. In this feasibility study, a (32)P-post-labeling thin-layer chromatography (TLC) assay is successfully used to measure AZT-DNA analogue and adducts formed in peripheral blood leukocytes of AZT treated patients. There are DNA analogue/adducts measured in all four AZT treated patients' DNA specimens. This assay is reliable with the significant coefficient of correlation in both intra-assay (r = 0.8761, P = 0.0001) and inter-assay (r = 0.8761, P = 0.0001). PMID- 15358302 TI - Quantitative determination of MK-0767, a dual alpha/gamma peroxisome proliferator activated receptor (PPAR) agonist, in human plasma by liquid chromatography tandem mass spectrometry. AB - 5-[2,4-Dioxothiazolidin-5-yl)methyl]-2-methoxy-N-[[(4-trifluoromethyl) phenyl]methyl]benzamide (I, MK-0767 or KRP-297, Fig. 1), is a dual alpha/gamma peroxisome proliferator-activated receptor (PPAR) agonist. A LC-MS/MS method for the determination of I in human plasma has been successfully developed, validated and applied to clinical programs. The analyte and internal standard (II) are extracted from 0.05 mL plasma via solid phase extraction (SPE). HPLC is used for the separation of I and II from possible co-extracted endogenous and other compounds. Detection is by MS/MS in multiple reaction monitoring (MRM) mode using a TurboIonSpray probe. The whole sample preparation is automated by using a Packard Multiprobe liquid handling system. The linear range is 4-2000 ng/mL in plasma. Recoveries were 71.1% and 69.4% for I and II, respectively. The method exhibited good linearity, reproducibility and sensitivity, selectivity and robustness when used for the analysis of clinical samples. PMID- 15358303 TI - Monolithic peptidyl sorbents for comparison of affinity properties of plasminogen activators. AB - Plasminogen activators are the proteases which convert plasminogen into plasmin dissolving, in its turn, the major component of blood clots, fibrin. They are extremely useful in heart attack therapy. Modern and most appropriate way of scaled up production of these valuable proteins is gene engineering. In this case, a separation and a purification of target product become the important steps of the whole process. Recently developed affinity chromatography on short monolithic columns seems to be a very attractive method for these purposes. High speed of a process prevents the protein's denaturation due to temperature or/and solvents influence. The better mass transfer mechanism (convection rather than diffusion) allows considering only biospecific complexing as time limiting step. Specificity of several synthetic peptides to plasminogen activators have been studied by affinity chromatography on short monolithic columns. Peptide ligands were synthesized by conventional solid phase peptide synthesis (SPPS). The immobilization procedure was carried out as a one step process at static conditions. The results of quantitative evaluation of such affinity interactions were compared with those established for plasminogen that is the natural affinity counterpart to both proteases. Additionally, some of investigated peptides were synthesized directly on GMA-EDMA disks and their affinity properties were compared with those established for the case of immobilized ligands. The possibility of using of synthetic peptidyl ligands for plasminogen activators isolation from native cell supernatant and model protein mixtures has been demonstrated. PMID- 15358304 TI - Determination of the new HIV-protease inhibitor atazanavir by liquid chromatography after solid-phase extraction. AB - An HPLC method previously described for the simultaneous assay of amprenavir, ritonavir, indinavir, saquinavir, nelfinavir and efavirenz is proposed here for the simultaneous analysis of the new HIV protease inhibitor atazanavir (ATV) in human plasma, by off-line solid-phase extraction (SPE) followed by HPLC coupled with UV-diode array detection. After viral inactivation by heat (60 degrees C for 60 min), plasma (600 microl) with clozapine (internal standard) is diluted 1 + 1 with phosphate buffer pH 7 and subjected to a SPE on a C18 cartridge. Matrix components are eliminated with 2 x 500 microl of a solution of 0.1% H(3)PO(4) neutralised with NaOH to pH 7. ATV is eluted with 3 x 500 microl MeOH. The resulting eluate is evaporated under nitrogen at room temperature and is reconstituted in 100 microl MeOH/H(2)O 50/50. A 40 microl volume is injected onto a Nucleosil 100-5 microm C18 AB column. ATV is analysed by UV detection at 201 nm using a gradient elution program with solvents constituted of MeCN and phosphate buffer adjusted to pH 5.14. The mobile phase also contains 0.02% sodium heptanesulfonate, enabling an excellent separation of ATV from the other HIV protease inhibitors (PIs) amprenavir, indinavir, saquinavir, ritonavir, lopinavir, nelfinavir and the non-nucleoside reverse transcriptase inhibitors (NNRTIs) efavirenz and nevirapine. The calibration curves are linear up to 10 microg/ml, with a lower limit of quantification of 0.2 microg/ml. The mean absolute recovery of ATV is 96.4 +/- 3.2%. The method is precise with mean inter day CVs within 1.1-6.1%, and accurate (range of inter-day deviations +0.3 to +2.3%). The method has been validated and is currently applied to the monitoring of ATV in HIV patients. PMID- 15358305 TI - Liquid chromatography-electrospray tandem mass spectrometric assay suitable for quantitation of halofuginone in plasma. AB - An LC-MS/MS method was developed to quantitate the potential antitumor agent halofuginone in plasma. The assay uses 0.2 ml of plasma; chlorohalofuginone internal standard; acetonitrile for protein precipitation; a Phenomenex SYNERGI 4 micro Polar RP 80A (4 microm, 100 mm x 2 mm) column; an isocratic mobile phase of methanol:water:formic acid (80:20:0.02, v/v/v); and positive-ion electrospray ionization with selective reaction monitoring detection. Halofuginone eluted at approximately 2.4 min, internal standard eluted at approximately 2.9 min, and no endogenous materials interfered with their measurement. The assay was accurate, precise, and linear between 0.1 and 100 ng/ml. Halofuginone could be quantitated in dog plasma for at least 24 h after an i.v. dose of 0.1mg/kg. The assay is being used in ongoing pharmacokinetic studies of halofuginone. PMID- 15358306 TI - Detection of cariogenic bacterial genes by microchip electrophoresis. AB - Allele-specific PCR primers were designed, based on the dextranase (dex) gene, to identify Streptococcus mutans and Streptococcus sobrinus in dental plaque; subsequently, PCR products were detected via microchip electrophoresis (ME). In order to amplify the dex gene fragment of S. mutans and S. sobrinus, the following two PCR methods were established. Duplex allele-specific PCR primers were designed on a region of low DNA homology; furthermore, 211 and 126-bp fragments were amplified for S. mutans and S. sobrinus, respectively. Common PCR primer for single allele-specific PCR was designed so as to sandwich a region exhibiting high homology and amplify PCR product of different DNA size due to deletion of small DNA fragment in two dex genes. S. mutans and S. sobrinus were amplified, leading to the generation of 202 and 226-bp products, respectively. Analysis of DNA base size by ME in order to achieve efficient separation employed a polymer mixture consisting of hydroxypropyl methylcellulose (HPMC) and polyethylene oxide (PEO). In the presence of a polymer mixture of 0.125% PEO/0.6% HPMC, two PCR products were obtained, displaying degree of separation of 226 bp/202 bp of 2.67 (Rs). Reproducibility (CV%, n = 7) was 0.3%; additionally, separation time was approximately 85 s. This method was applied to the detection of S. mutans and S. sobrinus in dental plaque. Detection of the dex genes of S. mutans and S. sobrinus characterized by quickness, precision and high sensitivity was possible. PMID- 15358307 TI - Determination of perfluorooctane sulfonate, perfluorooctanoate and perfluorooctane sulfonylamide in human plasma by column-switching liquid chromatography-electrospray mass spectrometry coupled with solid-phase extraction. AB - We report a method for determining fluorinated organic compounds such as perfluorooctane sulfonate (PFOS), perfluorooctanoate (PFOA) and perfluorooctanesulfonylamide (PFOSA) in human blood samples by column-switching liquid chromatography-electrospray mass spectrometry. The sample preparation prior to solid phase extraction (Waters Oasis HLB extraction column) involved simply mixing plasma sample with internal standard followed by centrifugation and extraction. The compounds were separated by reversed-phase chromatography with a C(8) column, and detected by mass spectrometry using selected ion monitoring in the negative mode. The average recoveries of PFOS, PFOA and PFOSA ranged from 82.2 to 98.7% (R.S.D.: from 2.0 to 5.2%, n = 6). The limits of quantitation of PFOS, PFOA and PFOSA at signal to noise (S/N = 10) were 0.5, 0.5 and 1.0 ng ml( 1). The method enables the precise determination of standards and can be applied to the detection of PFOS, PFOA and PFOSA in human plasma samples for monitoring human exposure. PMID- 15358308 TI - Single-step procedure for gas chromatography-mass spectrometry screening and quantitative determination of amphetamine-type stimulants and related drugs in blood, serum, oral fluid and urine samples. AB - We describe a rapid GC/MS assay for amphetamine-type stimulant drugs (ATSs) and structurally related common medicaments in blood, serum, oral fluid and urine samples. The drugs were extracted from their matrices and derivatized with heptafluorobutyric anhydride (HFBA) in a single step, using the following procedure: 100 microl (oral fluid) or 200 microl (blood, serum, urine) of the sample were mixed with 50 microl of alkaline buffer and 500 microl of extraction derivatization reagent (toluene + HFBA + internal standard), centrifuged, and injected into a GC/MS apparatus. As revealed by the validation data this procedure, with its limit of quantitation being set at 20 ng/ml for oral fluid, 25 ng/ml for blood or 200 ng/ml for urine, is suitable for screening, identification and quantitative determination of the ATSs and related drugs in all the matrices examined. Thus, time-consuming and expensive multiple analyses are not needed, unless specifically required. PMID- 15358309 TI - Characterizing complex peptide mixtures using a multi-dimensional liquid chromatography-mass spectrometry system: Saccharomyces cerevisiae as a model system. AB - A rugged, reproducible, multi-dimensional LC-MS system was developed to identify and characterize proteins involved in protein-protein interactions and/or protein complexes. Our objective was to optimize chromatographic parameters for complex protein mixture analyses using automated peptide sequence recognition as an analytical end-point. The chromatographic system uses orthogonal separation mechanisms by employing strong cation exchange (SCX) in the first dimension and reversed phase (RP) in the second dimension. The system is fully automated and sufficiently robust to handle direct injections of protein digests. This system incorporates a streamlined post analysis results comparison, called DBParser, which permitted comprehensive evaluation of sample loading and chromatographic conditions to optimize the performance and reproducibility. Peptides obtained from trypsin digestion of a yeast soluble extract provided an open-ended model system containing a wide variety and dynamic range of components. Conditions are described that resulted in an average (n = 4) of 1489 unique peptide identifications, corresponding to 459 non-redundant protein sequence database records (SDRs) in the 20 microg soluble fraction digest. PMID- 15358310 TI - Simultaneous determination of levofloxacin, gatifloxacin and moxifloxacin in serum by liquid chromatography with column switching. AB - Liquid chromatography with a column-switching technique was developed for simultaneous direct quantification of levofloxacin, gatifloxacin and moxifloxacin in human serum. Serum samples were injected on a LiChroCART 4-4 pre-column (PC) filled with a LiChrospher 100 RP-18, 5 microm where fluoroquinolones (FQs) were purified and concentrated. The FQs were back-flushed from the PC and then separated on a Supelcosil ABZ+ Plus (150 mm x 4.6 mm i.d.) analytical column with a mobile phase containing 10 mM phosphate buffer (pH 2.5), acetonitrile (88:12, v/v) and 2mM tetrabutyl ammonium bromide. The effects of ion-pair reagents, buffer type, pH and acetonitrile concentrations in the mobile phase on the separation of the three FQs were investigated. Fluorescence detection provided sufficient sensitivity to achieve a quantification limit of 125 ng/ml for levofloxacin and moxifloxacin; 162.5 ng/ml for gatifloxacin with a 5 microl sample size. The on-line process of extraction avoids time-consuming treatment of the samples before injection and run time is shortened. The recovery, selectivity, linearity, precision and accuracy of the method are convenient for pharmacokinetic studies or routine assays. PMID- 15358311 TI - Quantitative determination of the antitumor alkyl ether phospholipid edelfosine by reversed-phase liquid chromatography-electrospray mass spectrometry: application to cell uptake studies and characterization of drug delivery systems. AB - Edelfosine is a synthetic alkyl ether phospholipid that represents a promising class of antitumor agents. However, analytical methods to measure these type compounds are scarce. The lack of a reliable methodology to quantify edelfosine is a major problem in ongoing and scheduled preclinical and clinical trials with this drug. We evaluated the applicability of high-performance liquid chromatography-mass spectrometry to determine edelfosine in biological samples and polymeric delivery systems. Sample pre-treatment involved polymer precipitation or cell lysis with methanol. HPLC separation was performed on an Alltima RPC(18) narrow-bore column and edelfosine quantification was done by electrospray ionization mass spectrometry (ESI-MS) using positive ion mode and selected ion monitoring. Assays were linear in the tested range of 0.3-10 microg/ml. The limit of quantification was 0.3 ng/sample in both matrices, namely biological samples and polymeric delivery systems. The interassay precision ranging from 0.79 to 1.49%, with relative errors of -6.7 and 12.8%. Mean extraction recovery was 95.6%. HPLC-ESI-MS is a reliable system for edelfosine analysis and quantification in samples from different sources, combining advantages of full automation (rapidity, ease of use, no need of extensive extraction procedures) with high analytical performance and throughput. PMID- 15358312 TI - Determination of multiple redox-active compounds by high-performance liquid chromatography with coulometric multi-electrode array system. AB - Studies of the antioxidant defense system and the related metabolic pathways are often complicated by cumbersome analytical methods, which require separate and multi-step extraction and chemical reaction procedures. Further, assaying multiple parameters is limited because of the usual small sample amounts. High performance liquid chromatography coupled with a coulometric multi-electrode array system provides us high specificity and sensitivity to measure electrochemically oxidizable compounds in biological samples. In contrast to previously reported methods with two columns in series and a complex gradient elution profile, we have developed an automated procedure to simultaneously measure multiple redox-active low-molecular weight compounds that utilizes a single column with a simplified binary gradient profile. No other chemical reactions are necessary. In order to reduce the running time and yet achieve a reproducible retention time by the auto sampler injection, our gradient elution profile was modified to produce a shorter equilibration time, stable retention time, and a reduced cost per test. PMID- 15358313 TI - Determination of serotonin and its precursors in human plasma by capillary electrophoresis-electrospray ionization-time-of-flight mass spectrometry. AB - A specific capillary electrophoresis-time-of-flight mass spectrometry (CE-TOFMS) method for the determination of serotonin (5HT) and its precursors tryptophan (Trp) and 5-hydroxytryptophan (5HTP) in human platelet rich plasma is described. The analytes were removed from the plasma samples and preconcentrated by solid phase extraction (SPE) on mixed mode cation-exchange sorbents. The SPE recoveries were 71.6 +/- 3.1 for 5HT, 91.0 +/- 2.8 for Trp, and 95.3 +/- 5.9% for 5HTP. Deuterated analogues of 5HT and Trp were used as internal standards for quantitation purposes. Submicromolar detection limits were obtained for standard mixtures of all compounds and their deuterated isotopes, except 5HTP, which had detection limits in the low micromolar range. The potential usefulness of this method in the clinical setting was demonstrated by analyzing plasma extracts from healthy volunteers as well as from pathological samples. While 5HTP was not present in any of the analyzed samples, the levels of 5HT and Trp in both normal and pathological plasma were determined. PMID- 15358314 TI - Wide concentration range investigation of recovery, precision and error structure in liquid chromatography. AB - Using a typical HPLC assay, the characteristics of recovery, system precision and repeatability were investigated over a wide concentration range. In the presence of a constant amount of typical tablet excipients, the antidiabetic drug glibenclamide was analyzed in the range from 0.24 to 0.005mg/mL (18 concentration levels, 6 independent sample preparations each). On the basis of a typical concentration for an HPLC glibenclamide assay of 0.2mg/mL, this corresponds to a relative amount of 120-0.025% label claim. In the range from 120 to 0.075%, the recovery was found to be quite constant and systematically heightened mainly due to the evaporation from vials during centrifuging and the displacement of solvent volume by the added matrix. Both system precision and repeatability remain almost constant in the interval from 120 to 10% at a R.S.D.% of 0.31 and 0.70%, respectively, indicating that the sample preparation is the major error source in this range (0.63%). Between 10 and 0.25%, a linear relationship between the logarithmized concentration and the repeatability was noted. However, for lower amounts close to the limit of quantitation, the R.S.D.% of measurements increases much more distinctly. This increase is caused by a strong rise of the system precision. At this concentration range, system precision and repeatability are not significantly different any longer. This leads to the conclusion that with the injection error being constant the peak integration error becomes the dominating error source at low concentrations, e.g. at concentrations below the five-fold of the LOQ. The results obtained here agree well with earlier published data. As the quantitation limit of 0.05% can be regarded as typical for a pharmaceutical impurity control test, generalizations of these findings from this extensive data set should be possible. In this context, peak integration and improvements of the signal-to-noise ratio are the most promising measures to improve an unsatisfactory precision in LC. PMID- 15358315 TI - Micro-quantitation of lipids in serum-free cell culture media: a critical aspect is the minimization of interference from medium components and chemical reagents. AB - Lipids (fatty acids) at a concentration range of 10-100 microg/L are essential components included in most serum-free cell culture medium formulations. A gas chromatography/mass spectrometry (GC/MS) method for the micro-quantitation of lipids, determined as fatty acid methyl esters (FAMEs), in complex serum-free cell culture media was developed. The interference of derivatizing reagents, extraction solvents and medium additives in the micro-quantitation of lipids was also examined. The results show that the concentration of fatty acids such as palmitic and stearic acids detected in derivatizing reagents or extraction solvents was in the range of 10-230 microg/L. Tween-80, a surfactant and medium additive, produced nearly 20 FAMEs alone when methylated using a derivatizing agent. Moreover, the surfactant Pluronic F-68, a medium additive, interfered in the FAME recovery. Procedures, which include use of low volumetric ratio of reagent to medium and precipitation of the above surfactants, were developed to minimize background FAMEs to levels which do not significantly affect the quantitation of medium lipids and to diminish the interference caused by Pluronic F-68. Fatty acid concentrations in several complex serum-free culture media were quantitated by this method and were very close to the values indicated in their formulations. PMID- 15358316 TI - Development and validation of a liquid chromatographic-tandem mass spectrometric method for the determination of caffeic acid phenethyl ester in rat plasma and urine. AB - Caffeic acid phenethyl ester (CAPE) is one of the most bioactive compounds of propolis, a resinous substance collected and elaborated by honeybees. A new liquid chromatography-electrospray ionisation tandem mass spectrometric method was developed and validated for its determination in rat plasma and urine, using taxifolin as internal standard. After sample preparation by liquid/liquid extraction with ethyl acetate, chromatographic separations were carried out with an ODS-RP column using a binary mobile phase gradient of acetonitrile in water. Detection was performed using a turboionspray source operated in negative ion mode and by multiple reaction monitoring. The method was validated, showing good selectivity, sensitivity (LOD = 1 ng/ml), linearity (5-1000 ng/ml; r > or = 0.9968), intra- and inter-batch precision and accuracy (< or =14.5%), and recoveries (94-106%) in both plasma and urine. Stability assays have shown that CAPE is rapidly hydrolysed by plasmatic esterases, which are however inhibited by sodium fluoride. The method was applied to the determination of CAPE levels in rat plasma and urine after oral administration, showing that CAPE is rapidly absorbed and excreted in urine both as unmodified molecule and as glucuronide conjugate. PMID- 15358317 TI - Electrochemiluminescence detection with integrated indium tin oxide electrode on electrophoretic microchip for direct bioanalysis of lincomycin in the urine. AB - In this article, an antibiotic, lincomycin was determined in the urine sample by microchip capillary electrophoresis (CE) with integrated indium tin oxide (ITO) working electrode based on electrochemiluminescence (ECL) detection. This microchip CE-ECL system can be used for the rapid analysis of lincomycin within 40s. Under the optimized conditions, the linear range was obtained from 5 to 100 microM with correlation coefficient of 0.998. The limit of detection (LOD) of 3.1 microM was obtained for lincomycin in the standard solution. We also applied this method to analyzing lincomycin in the urine matrix. The limit of detection of 9.0 microM was obtained. This method can determine lincomycin in the urine sample without pretreatment, which demonstrated that it is a promising method of detection of lincomycin in clinical and pharmaceutical area. PMID- 15358318 TI - Determination of glimepiride in human plasma using semi-microbore high performance liquid chromatography with column-switching. AB - A fully automated semi-microbore high performance liquid chromatographic (HPLC) method with column-switching using UV detection was developed for the determination of glimepiride from human plasma samples. Plasma sample (900 microl) was deproteinated and extracted with ethanol and acetonitrile. The extract (70 microl) was directly injected into a Capcell Pak MF Ph-1 pre-column where the primary separation occurred to remove proteins and retain drugs using a mixture of acetonitrile and 10mM phosphate buffer (pH 2.18) (20:80, v/v). The analytes were transferred from the pre-column to an intermediate column using a switching valve and then subsequently separated on an analytical column and monitored with UV detection at 228 nm. Glimepiride was eluted with retention time 34.9 min without interference of endogenous substance from plasma. The limit of quantification (LOQ) was 10 ng/ml for glimepiride. The calibration curves were linear over the concentration range of 10-400 ng/ml (r(2) = 0.9997). Moreover, inter- and intra-day precisions of the method were less than 15% and accuracies were higher than 99%. The developed method was successfully applied for the quantification of glimepiride in human plasma and was used to support a human pharmacokinetic study following a single oral administration of 2 mg glimepiride. PMID- 15358319 TI - Analysis of non-covalent aggregation of synthetic hPTH (1-34) by size-exclusion chromatography and the importance of suppression of non-specific interactions for a precise quantitation. AB - There are few methods available for the rapid and precise quantitation of non covalent aggregation. Size-exclusion chromatography (SEC), a traditional approach, used to measure the non-covalent aggregation can easily disrupt the weak forces holding an aggregate together. Under the conditions described in this paper the disaggregation of non-covalent aggregate of the synthetic human parathyroid hormone hPTH (1-34) due to hydrophobic/electrostatic interactions with the size-exclusion chromatography column packing was completely suppressed. This report details the effectiveness of adding salts and organic solvents in the mobile phase to overcome non-specific interactions that disrupt the aggregate during the SEC process and may aid in the understanding precise quantitation of non-covalent aggregation. PMID- 15358320 TI - Urinary analysis of 16(5alpha)-androsten-3alpha-ol by gas chromatography/combustion/isotope ratio mass spectrometry: implications in anti doping analysis. AB - We present a method for the analysis of urinary 16(5alpha)-androsten-3alpha-ol together with 5beta-pregnane-3alpha,20alpha-diol and four testosterone metabolites: androsterone (Andro), etiocholanolone (Etio), 5alpha-androstane 3alpha,17beta-diol (5alphaA), 5beta-androstane-3alpha,17beta-diol (5betaA) by means of gas chromatography/combustion/isotopic ratio mass spectrometry (GC/C/IRMS). The within-assay and between-assay precision S.D.s of the investigated steroids were lower than 0.3 and 0.6 per thousand, respectively. A comparative study on a population composed of 20 subjects has shown that the differences of the intra-individual delta(13)C-values for 16(5alpha)-androsten 3alpha-ol and 5beta-pregnane-3alpha,20alpha-diol are less than 0.9 per thousand. Thereafter, the method has been applied in the frame of an excretion study following oral ingestion of 50 mg DHEA initially and oral ingestion of 50mg pregnenolone 48 h later. Our findings show that administration of DHEA does not affect the isotopic ratio values of 16(5alpha)-androsten-3alpha-ol and 5beta pregnane-3alpha,20alpha-diol, whereas the isotopic ratio values of 5beta-pregnane 3alpha,20alpha-diol vary by more 5 per thousand upon ingestion of pregnenolone. We have observed delta(13)C-value changes lower than 1 per thousand for 16(5alpha)-androsten-3alpha-ol, though pregnenolone is a precursor of the 16-ene steroids. In contrast to 5beta-pregnane-3alpha,20alpha-diol, the 16-ene steroid may be used as an endogenous reference compound when pregnenolone is administered. PMID- 15358321 TI - Study on the precipitation reaction between baicalin and berberine by HPLC. AB - The solubility product equilibrium constant for the precipitation between baicalin and berberine was investigated because of the current interest in decocting process of complex prescription of Chinese herbal medicine. HPLC was used for determining two compounds' equilibrium concentrations at different precipitate conditions to calculate thermodynamic constants and study kinetic process. The analysis was performed on a Kromasil C(18) column with TEA-adjusted 0.02 mol/L H(3)PO(4) (pH 4.82)-acetonitrile (75:25) as mobile phase at a flow rate of 1.0 ml/min, with detection at 254 nm. According to the experiment result, the molar ratio of baicalin and berberine in precipitate is about 1:1. The experimental K(sp) values are (1.01 +/- 0.12) x 10(-9)mol(2)/L(2) at 20 degrees C in 0.02 mol/L NaH(2)PO(4)-Na(2)HPO(4) (pH 4.82), and (3.20 +/- 0.46) x 10( 9)mol(2)/L(2) at 40 degrees C in the same buffer. The precipitate reaction is an exothermic process and occurs immediately, even though the precipitate cannot be observed in time because the precipitate is light, yellow, flocculous and suspending in the yellow solution. PMID- 15358322 TI - Simple and sensitive fluorimetric liquid chromatography method for the determination of valproic acid in plasma. AB - A simple and sensitive liquid chromatographic method is described for the analysis of valproic acid in human plasma. The method is based on the derivatization of valproic acid extracted from acidified plasma with 2-(2 naphthoxy)ethyl 2-(piperidino)ethanesulfonate. The resulting derivative is highly responsive to a fluorimetric detector (excitation at 230 nm and emission at 350 nm), giving a low detection limit of 0.6 microM (S/N = 3, 10 microl injected). The relative standard deviations of the method for intra- and inter-day analyses (n = 5) are below 3.3 and 4.1%, respectively. Toluene was used for the extraction of valproic acid from plasma and the toluene extract obtained was subjected to subsequent derivatization without solvent replacement. The simple method was applied to the analysis of valproic acid in plasma of dosed patients using only small amount of sample (10-50 microl plasma). PMID- 15358323 TI - Mesenteric lymph collected during peritonitis or sepsis potently inhibits gastric motility in rats. AB - Gastrointestinal motility is strongly inhibited during peritonitis or sepsis and proinflammatory cytokines released into mesenteric lymph during an acute gastrointestinal insult mediate systemic responses. We investigated whether mesenteric lymph collected during peritonitis or sepsis inhibits gastric motility and gastric emptying. Mesenteric lymph was collected for 12 hours from three experimental groups: vehicle (saline, 1 ml, intraperitoneally [ip], control lymph), peritonitis (0.5% acetic acid, 1 ml, ip, peritonitis lymph), and sepsis (lipopolysaccharide [LPS], 5 mg/kg, 1 ml, ip, sepsis lymph). Gastric motility and gastric emptying were measured in recipient rats in response to lymph injections into the jugular vein. Quantitative polymerase chain reaction (PCR) for tumor necrosis factor alpha (TNFalpha) gene expression in the jejunum and in lymph cells were measured during sepsis. Mesenteric lymph flow significantly increased during peritonitis or sepsis (lymph flow [ml] per 60 minutes; control 2.45 +/- 0.04; peritonitis 2.67 +/- 0.07; sepsis 3.25 +/- 0.1, p < 0.01 vs. control). Injection of peritonitis or sepsis lymph (1 ml) produced a significant and prolonged inhibition of gastric motility in recipient rats (decrease in intragastric pressure and duration: control lymph -0.14 +/- 0.05 cm H(2)O, 1.89 +/- 1.31 minutes; peritonitis lymph: -0.56 +/- 0.06 cm H(2)O, 9.9 +/- 0.9 minutes; sepsis lymph: -0.51 +/- 0.05 cm H(2)O, 6.9 +/- 0.6 minutes; p < 0.001 vs. control for all comparisons). Gastric emptying was significantly inhibited by continuous infusion of sepsis lymph (3 ml per 60 minutes; gastric emptying: saline 81% +/- 4%; control lymph: 80% +/- 6%; sepsis lymph: 44% +/- 10%; p < 0.001 vs. control). TNFalpha gene expression in the gut wall of the jejunum increased during sepsis over 90-fold within the first 2 hours and decreased continuously thereafter (relative TNFalpha mRNA transcription: basal 1.0 +/- 0.05; LPS 2 hours: 91.9 +/- 2.6, p < 0.001 vs. basal; 12 hours: 24.7 +/- 16.8, not significant [NS]; 24 hours: 7.0 +/- 3.4, NS). In conclusion, mediators in mesenteric lymph, possibly cytokines, may be responsible for the inhibition of gastric motility during peritonitis or sepsis. Because the composition of mesenteric lymph probably reflects the interstitial fluid of the gut wall, monitoring visceral lymph might be an extremely beneficial tool to determine mediators released during impaired gut wall function. PMID- 15358324 TI - Survival and functional quality of life after resection for hepatic carcinoid metastasis. AB - Retrospective studies suggest that resection improves 5-year survival for patients with hepatic carcinoid metastasis (HCM). The purpose of our study was to describe clinical outcomes following resection for HCM, including survival and longitudinal functional quality of life (QOL). We reviewed the records of patients undergoing resection for HCM from 1980 to 2001 at our institution. Outcome measures included tumor symptoms, biochemical tumor markers, functional QOL through Karnofsky functional scores, and survival. Thirteen patients underwent a total of 17 resections. Overall 5-year survival was 85%. Eleven patients were symptomatic, including eight with classic carcinoid syndrome. Nine experienced complete relief of symptoms and two had incomplete relief for 30 +/- 12 months. Eight patients had elevated tumor markers, and 50% of these had postoperative normalization of all tumor markers that persisted to the close of the study. For the 10 patients with longitudinal follow-up available to 54 months, significant improvement in functional QOL was observed at all follow-up time points compared to preresection functional QOL (P < 0.05). Resection of >/=90% tumor volume was significantly associated with more favorable survival and tumor marker normalization compared to resection of <90% tumor volume (P < 0.01 and P < 0.05, respectively), but trajectory of functional QOL improvement did not differ between these two groups (P=0.24). We conclude that resection for HCM is associated with significantly improved and sustained functional QOL and prolonged survival. Resection of >/=90% tumor volume is significantly associated with extended survival and normalization of tumor markers, but is not required for symptomatic or functional QOL improvement. PMID- 15358325 TI - Real-time spectroscopic assessment of thermal damage: implications for radiofrequency ablation. AB - Radiofrequency ablation (RFA) is an evolving technology used to treat unresectable liver tumors. Currently, there is no accurate method to determine RFA margins in real-time during the procedure. We hypothesized that a fiber-optic based spectroscopic monitoring system could detect thermal damage from RFA in real-time. Fluorescence (F) and diffuse reflectance (Rd) spectra were continuously acquired from within the expected ablation zone during canine hepatic RFA using a fiber-optic microinterrogation probe (MIP). The F and Rd spectral feedback were continuously monitored and ablations were stopped based on changes in spectra alone. After each ablation, the MIP tract was marked with India ink and the ablation zone was excised. The relationship of the MIP to the zone of ablation was examined grossly and microscopically. F and Rd spectral changes occurred in three characteristic phases as the ablation zone progresses past the MIP. Phase 1 indicates minimal deviation from normal lives. Phase 2 occurs as the MIP lies within the hemorrhagic zone of the ablated tissue. Phase 3 correlates with complete tissue coagulation. The absolute magnitude of spectral change correlates with the gross and histologic degree of thermal damage. Optical spectroscopy is a technology that allows real-time detection of thermal tissue damage. In this study, both F and Rd spectroscopy accurately defined the advancing hemorrhagic edge of the zone of ablation and the central coagulation zone. These results suggest that F and Rd spectroscopy can be used to create a real-time feedback system to accurately define RFA margins. PMID- 15358326 TI - Laparoscopic ventral incisional hernia repair: a more effective alternative to conventional repair of recurrent incisional hernia. AB - Conventional repair of recurrent ventral incisional hernia is associated with a higher recurrence rate (30%-50%) than repair of primary incisional hernia (11% 20%). Laparoscopic incisional hernia repair (LIHR) can significantly reduce the recurrence rate of primary hernia to less than 5%. In this study, we evaluate the efficacy of repairing recurrent incisional hernia laparoscopically. One-hundred and seventy consecutive patients undergoing LIHR between January 1995 and December 2002 were prospectively reviewed. Patients with recurrent incisional hernia (n=69) were compared to patients with primary incisional hernia (n=101). Patient demographics and perioperative and postoperative data were recorded prospectively. Follow-up was obtained from office visits and telephone interviews. Statistical analysis was performed using the Student t test and the chi(2) test. Results are expressed as means +/- standard deviation. The patients with recurrent incisional hernia had a mean of 1.9 +/- 1.3 previous repairs, higher body mass index (BMI) (34 +/- 6 kg/m(2) vs. 33 +/- 8 kg/m(2), P=0.46), larger defect size (123 +/- 115 cm(2) vs. 101 +/- 108 cm(2), P=0.06), and longer operative time (119 +/- 61 minutes vs. 109 +/- 44 minutes, P=0.11). The complication rate was higher in the recurrent group (28% vs. 11%, P=0.01), but the recurrence rate was not different (7% vs. 5%, P=0.53). The mean time to recurrence was significantly shorter in the recurrent group (3 +/- 2 months vs. 14 +/- 7 months, P < 0.0001). The mean follow-up interval was 19 +/- 18 months in the recurrent group and 27 +/- 20 months in the primary group. Although laparoscopic repair of recurrent incisional hernia resulted in a higher recurrence and complication rate than laparoscopic repair of primary incisional hernia, the rates were lower than those reported for conventional repair of recurrent incisional hernia. Laparoscopic repair of recurrent incisional hernia is an effective alternative to conventional repair. PMID- 15358327 TI - Colonic interposition vs. gastric pull-up after total esophagectomy. AB - Gastric pull-up is the most frequent reconstruction after esophagectomy. In this report we aimed to compare gastric pull-up with colonic interposition in terms of graft function and patient satisfaction. Of 62 patients undergoing esophagectomy, reconstruction was performed by colonic interposition in 11 and gastric pull-up in 51 (without pyloric drainage in 44 and with pyloric drainage in 7). All esophagectomies were performed transhiatally. Patient follow-up ranged from 6 to 132 months (median 14 months). Follow-up examinations were performed 1, 9, 15, and 24 months postoperatively. The following factors were evaluated: time to the start of oral liquid and solid nutrients without vomiting, frequency of regurgitation, presence of pillow staining (night regurgitation), postprandial fullness, and degree of satisfaction during and after eating compared between groups undergoing colonic interposition and gastric pull-up with or without pyloric drainage. Among patients undergoing gastric pull-up, regurgitation was observed in 22% to 27% during follow-up. None of the patients with colonic interposition had reflux or regurgitation. Twenty-five percent of patients with gastric pull-up without drainage and 66% of patients with gastric pull-up plus drainage had reflux esophagitis at 15 months. No esophagitis was observed in patients with colonic interposition during the same period. Overall satisfaction was superior in patients undergoing colonic interposition followed by gastric pull-up with no drainage. Colonic interposition after esophageal resection is a viable option. Our study suggests that function of the replacement is better in this group of patients. PMID- 15358328 TI - Major bile duct injuries after laparoscopic cholecystectomy: a tertiary center experience. AB - Bile duct injury is a severe and potentially life-threatening complication of laparoscopic cholecystectomy. Several series have described a 0.5% to 1.4% incidence of bile duct injuries during laparoscopic cholecystectomy. The aim of this study was to report on an institutional experience with the management of complex bile duct injuries and outcome after surgical repair. Data were collected prospectively from 40 patients with bile duct injuries referred for surgical treatment to our center between April 1998 and December 2003. Prior to referral, 35 patients (87.5%) underwent attempts at surgical reconstruction at the primary hospital. In 77.5% of the patients, complex type E1 or type E2 BDI was found. Concomitant with bile duct injury, seven patients had vascular injuries. Roux-en Y hepaticojejunostomy was carried out in 33 patients. In two patients, Roux-en-Y hepaticojejunostomy and vascular reconstruction were necessary. Five patients, one with primary nondiagnosed Klatskin tumor, required right hepatectomy. Two patients, both with bile duct injuries and vascular damage, died postoperatively. Because of progressive liver insufficiency, one of them was listed for high urgency liver transplantation but died prior to intervention. At the median follow-up of 589 days, 82.5% of the patients are in excellent general condition. Seven patients have signs of chronic cholangitis. Major bile duct injuries remain a significant cause of morbidity and even death after laparoscopic cholecystectomy. Because they present a considerable surgical challenge, early referral to an experienced hepatobiliary center is recommended. PMID- 15358329 TI - Extrahepatic bile duct resection in combination with liver resection for hilar cholangiocarcinoma: a report of 42 cases. AB - From September 1986 until December 2001, 42 patients (20 males and 22 females) underwent a combined extrahepatic bile duct resection (EHBDR) and liver resection (LR) for hilar cholangiocarcinoma (HC). The aim of this study was to analyze patient survival, morbidity, and mortality as well as to seek predictive factors. The 1-, 3-, and 5-year actuarial patient survival was 72%, 37%, and 22%, respectively. Median survival was 19 months. Hospital mortality, all due to septic complications, was 12%. Morbidity was observed in 32 patients (76%). Infections were the most dominant complication. Patients (n=11) with American Joint Committee on Cancer (AJCC) stage I or stage II tumors exhibited a superior survival compared with patients (n=31) with stage III or IV tumors (p=0.023). Patients with tumor-free lymph nodes (n=26) indicated a greater survival compared with patients with tumor-positive lymph nodes (n=16) (p=0.004). Patients undergoing vascular reconstructions indicated a trend toward higher mortality and lower survival (p=0.068). Over 20% of the patients with hilar cholangiocarcinoma can survive more than 5 years after a combined EHBDR and LR at the cost of 12% perioperative mortality and a 76% morbidity. Results might improve with the prevention of infectious complications and improved selection of patients to avoid vascular reconstruction and to predict a negative nodal state. PMID- 15358330 TI - Hepatocellular ultrastructure after ischemia/reperfusion injury in human orthotopic liver transplantation. AB - The number of patients requiring organ transplants still outpaces the number of available transplantable organs. During the process of orthotopic liver transplantation (OLTx), donor organs undergo significant stress resulting from ischemia and reperfusion. Healthy organs respond to this stressful environment with compensatory mechanisms that ideally allow for complete recovery. However, "marginal" organs do not compensate as well. Hepatic steatosis typically renders an organ nontransplantable; a liver with 30% or more fat has a 25% chance of primary nonfunction (PNF) or graft failure after a technically sound operation. In this study, we report on the significant markers of cellular ultrastructural change in steatotic livers. These include glycogen content, mitochondrial swelling, and hepatocellular blebbing. The data disclosed here argue that further investigation of these factors in marginal organs subjected to I/R may better facilitate our understanding of PNF. PMID- 15358331 TI - Isolated Roux-loop pancreaticojejunostomy: a series of 61 patients with zero postoperative pancreaticoenteric leaks. AB - There have been approximately 70 reported variations of reconstruction after pancreaticoduodenectomy (PD). The pancreaticojejunal (PJ) anastomosis is the source of most reported morbidity and mortality. In this study, we aimed to identify the anastomotic leak rate in patients undergoing PD for malignant disease using a proximal isolated jejunal pancreatic anastomosis. Sixty-one consecutive patients undergoing PD (26 women and 35 men; age range, 41-79 years, mean age, 62 years). had an identical reconstruction. The PJ anastomosis was performed using the most proximal isolated jejunum in two layers: interrupted 4.0 Prolene was used to achieve mucosal/ductal continuity, and 3.0 Prolene was used for the serosal/parenchymal anastomosis, around an appropriately sized stent. All postoperative complications were recorded. A pancreatic leak was defined as persistent discharge of amylase-rich pancreatic drain fluid. The overall complication rate was 44% (27 of 61, including 15 chest infections, 8 wound infections, and 2 postoperative cardiac arrhythmias). There were 3 deaths (30-day mortality rate, 5%). One patient died after a cerebrovascular accident, one from respiratory failure secondary to pneumonia, and the third of methicillin resistant Staphylococcus aureus septicemia after small bowel ischemia caused by pressure necrosis from a drain. There were no PJ anastomotic leaks. This method of pancreatojejunostomy has produced a 0% leak rate in this center. PMID- 15358332 TI - Clinical characteristics, treatment, and outcome of pancreatic Schwannomas. AB - This article involves the study of a patient with a rare benign schwannoma in the body of the pancreas. After reviewing 39 patient cases previously reported in the literature, a discussion of the schwannoma with regard to clinical presentation, diagnosis, and treatment is examined. A review of the patient's chart was performed along with a review of the literature using a Medline search. Translations were performed whenever necessary. There are 23 reports of 29 patient cases of pancreatic schwannomas in English and European literature and one report of 10 patient cases in the Japanese literature. The mean age was 57.75 years (range 32-89) and the male-to-female (M:F) ratio was 17:23. The mean reported size was 8.79 cm. The lesion was located in the head in 16 patients (40%), the body in 8 patients (20%), the body and tail in 8 patients (20%), the tail in 6 patients (15%), the head and body in 1 patient (2.5%), and the location was not specified in 1 patient (2.5%). Of the English and European patients, 11 out of 30 patients (36.7%) exhibited solid tumors and 14 out of 30 patients (46.7%) exhibited cystic tumors. The majority of the tumors (35 out of 40) were benign, but there were five reported malignancies. There were no deaths or recurrences reported with a follow-up of 18.68 months +/- 24.09 (range 3-108 months). Pancreatic schwannomas are rare, and the preoperative diagnosis is difficult. Intraoperative frozen section can confirm the diagnosis of a benign schwannoma. Enucleation of the tumor from the surrounding parenchyma is recommended, if possible. Patients undergoing resection indicate an excellent long-term prognosis. PMID- 15358333 TI - Pancreatic head resection with segmental duodenectomy for intraductal papillary mucinous tumors of the pancreas. AB - Various modifications of organ-preserving pancreatic resections have been performed for intraductal papillary mucinous tumor (IPMT) of the pancreas. The aim of this study was to evaluate usefulness of pancreatic head resection with duodenal segmentectomy (PHRSD), which is one of the organ-preserving pancreatic resections for IPMT. Pancreatic head resection with duodenal segmentectomy was indicated for the branch duct type of IPMT. Eight patients underwent PHRSD. The mean operative time was 390 minutes, and the mean blood loss was 1270 ml. Duodenal ischemia was prevented by preserving the duodenal branches of the gastroduodenal artery and the anterior inferior pancreaticoduodenal artery. Complications occurred in four patients: one with pancreatic leak, one with choledochoduodenal anastomotic stenosis, and two with delayed gastric emptying. However, no deaths occurred. The final pathologic diagnosis was adenoma in seven patients and carcinoma in situ in one patient. Six of eight patients had an adenoma with papillary growth in the main pancreatic duct. Postoperative pancreatic endocrine and exocrine functions were satisfactory. All patients were alive without recurrent disease at a median follow-up of 30 months. Pancreatic head resection with duodenal segmentectomy appears to be a useful procedure as an organ-preserving pancreatic resection for the branch duct type of IPMT, because this procedure allows a safe and complete resection of the pancreatic head without ischemia of the common bile duct and the duodenum. PMID- 15358334 TI - Early results of one-year robotic surgery using the Da Vinci system to perform advanced laparoscopic procedures. AB - Robotic technology has recently been introduced to gastrointestinal laparoscopic surgery. We prospectively evaluated early results of robotic surgery using the Da Vinci system in our department. Data were prospectively collected in 40 patients who underwent robotic surgery during a 1-year period. We performed 3 cholecystectomies, 10 anterior fundoplications for gastroesophageal reflux disease, 17 transperitoneal adrenalectomies, 2 Heller myotomies, 5 procedures for rectal prolapse, and 3 colpohysteropexies for genital prolapse. The results for robotic adrenalectomies and anterior fundoplications were compared with the results from patients who underwent these procedures laparoscopically without robotic assistance at our department during the same period. We encountered two conversions to laparotomy (5%) and one conversion to standard laparoscopy (2.5%). There was no morbidity imputable to the robotic approach and no deaths. The mean operative times were significantly longer in robotic groups compared with laparoscopic groups for adrenalectomies and fundoplications. The Da Vinci robotic system enables surgeons to perform advanced laparoscopic procedures with ease, safety, and precision. We believe that preferable indications for using this system are to perform surgery in narrow spaces (pelvic surgery) or when precise dissection is mandatory (Heller myotomy). PMID- 15358335 TI - Gastric cancer surgery without drains: a prospective randomized trial. AB - Prophylactic drain placement during major abdominal surgery has been widely practiced without clear scientific evidence to support it. We hypothesized that prophylactic drain placement is not necessary in gastric cancer surgery. A randomized prospective trial was conducted between February 1, 2001, and July 30, 2001. Patients were randomly assigned to either the drain group or the no-drain group. One hundred seventy patients completed the study by undergoing either subtotal or total gastrectomy with D2 lymph node dissection. Surgical outcome between the two groups was compared within the subtotal and total gastrectomy subgroups. Postoperative complication within 30 days was the primary end point of the study. No significant difference was noted in the incidence of postoperative complication between the drain group and the no-drain group. The results of this study suggest that prophylactic drain placement does not offer additional benefit for patients undergoing gastric cancer surgery with extended lymph node dissection. PMID- 15358336 TI - Pancreaticoduodenectomy. PMID- 15358337 TI - Current management of gastrointestinal carcinoid tumors. AB - Gastrointestinal carcinoid tumors are rare neuroendocrine tumors arising from the embryologic primitive gut. Depending on the location in the gastrointestinal tract, these tumors may secrete a variety of hormonally active substances. However, many of these tumors are found incidentally, or the diagnosis is made postoperatively. Also, there is a significant incidence of multicentric carcinoid tumors and synchronous noncarcinoid malignancies in these patients. Treatment is usually based on the size of the tumor. Surgical resection remains the cornerstone of therapy. For advanced metastatic disease, somatostatin analog therapy and surgical debulking provide the best symptomatic relief and may improve survival. Recent studies have demonstrated a benefit from radiolabeled somatostatin analogs for carcinoid tumor localization. In contrast, radiolabeled somatostatin analogs have shown little therapeutic benefit. Future directions include somatostatin receptor profiling of carcinoid tumors, with somatostatin analog therapy targeting the specific receptors. PMID- 15358338 TI - Implications of proprotein Convertase 5 (PC5) in the arterial restenotic process in a porcine model. AB - INTRODUCTION: Convertases (PCs), especially PC5, have been detected in various layers of atherosclerotic and injured arteries. We postulate that PCs could be important enzymes in vascular disease thus studied PC5 expression in a porcine balloon and stent coronary arterial vascular injury model. METHODS: Immunohistochemistry and in situ hybridization of slides of porcine arteries from paraffin blocks were studied 1, 7, 14 and 28 days post injury. RESULTS: Immunohistochemistry studies show expression of PC5 in control artery endothelial cells, weak medial smooth muscle cell (SMC) staining and strong staining in the small nerves of the adventitia. At 7, 14 and 28 days postinjury, there is strong positive PC5 staining of the neointimal cells and the adventitial vasa vasora and myofibroblasts. Colocalization immunohistochemistry confirms the smooth muscle staining properties of the myofibroblast-like cells in both these locations. Single-label immunohistochemistry studies show the same cells to stain strongly positive with TGF-B, PDGF, matrix metalloproteinase-2 (MMP-2) and MMP-9. CONCLUSION: PC5 may be involved in the process of arterial injury via its effect on growth factors (GFs) and mediators. These preliminary observations suggest that the convertases, especially PC5, represent a target for future study in the process of arterial injury. PMID- 15358339 TI - Perivascular injury leads to a reduction in vascular reactivity of the collared and to an enhancement on contralateral carotid artery of rats. AB - BACKGROUND: The first response to perivascular injury is observed in the adventitial layer. The purpose of this study was to determine the time course of vascular reactivity alterations after collar injury in rats. We also analyzed the relation between adventitial layer injury and vascular responsiveness to vasoconstrictor agents. METHODS: Wistar rats had a silicone collar positioned around the carotid artery. The ipsilateral and contralateral arteries were morphologically analyzed 4, 7, 14 and 28 days after injury, and cumulative concentration-response curves to phenylephrine (Phe), angiotensin II (Ang II) and KCl were obtained for arteries isolated from collared and sham-operated groups. RESULTS: Inflammatory cells and fibroblasts were observed in the adventitial layer of collared arteries 4, 7, 14 and 28 days after injury. Intimal thickening was observed in collared arteries only 14 and 28 days after perivascular injury. A decrease in maximum effect values (Emax) for Phe, Ang II and KCl was observed in the collared artery when compared with the contralateral artery at all times after injury, whereas an increase in vascular responsiveness was observed in the contralateral artery 4 days after injury. CONCLUSIONS: The impairment of the contractile response preceded the intimal thickening. The compromise of vascular reactivity coincided with the presence of inflammatory cells and angiogenesis in the adventitial layer. The enhancement of the efficacy and potency of Ang II and Phe in collared-contralateral arteries 4 days after collar placement may be related to a receptor-mediated compensatory mechanism stimulated by the collar injury. PMID- 15358340 TI - The natural history of collagen and alpha-actin expression after coronary angioplasty. AB - BACKGROUND: Restenosis after percutaneous transluminal coronary angioplasty (PTCA) is a result of remodeling and deposition of new mass. The new mass is formed by invading and replicating cells and by extracellular matrix (ECM), of which collagens constitute the dominating component. Smooth muscle actin is an important element in cell contraction. We tested the hypothesis that the accumulation of collagen and actin correlates with the development of postinjury luminal narrowing. METHODS AND RESULTS: Thirty-five pigs underwent balloon angioplasty and were killed 0, 1, 4, 7, 14, 28, and 56 days later. Tissue samples from the left circumflex artery were in paraffin, sectioned, and immunostained for Collagen Types I and III and alpha1-smooth muscle actin. Collagen accumulation was measured separately in intima, media, and adventitia using computerized semiautomatic planimetry. The injury produced a strong healing response, with a marked accumulation of collagen in all three vessel wall layers. However, the accumulation in adventitia began surprisingly early (1 to 4 days after PTCA) and stopped at Day 7, i.e., before luminal narrowing occurred (14 to 28 days after PTCA in our model). Furthermore, a conspicuous accretion of collagen occurred in the injured area of the medial layer. This response attenuated 14 days after PTCA. Neointimal collagen accumulation took place parallel to neointima formation 2 to 4 weeks after injury. Extramedial smooth muscle actin occurred predominantly from Days 4 to 14 in neointima. Only small quantities of actin were observed in the (neo-)adventitia. Furthermore, adventitial actin was a temporary phenomenon that disappeared between Days 14 and 28. CONCLUSION: Adventitial and medial collagen deposition apparently occurs before luminal narrowing, indicating that the bulk of new mass in adventitia and media is not the cause of vessel remodeling, but possibly stabilizes the vessel wall and impairs compensatory outward remodeling. The accumulation of actin positive cells and collagen takes place in neointima parallel to luminal narrowing, which suggests that a contraction within the neointimal mass may contribute to the remodeling process. PMID- 15358341 TI - Decreased numbers of T-lymphocytes and predominance of recently recruited macrophages in the walls of peripheral pulmonary arteries from 26 patients with pulmonary hypertension secondary to congenital cardiac shunts. AB - INTRODUCTION: In the primary form of pulmonary hypertension (PH), the involvement of inflammation in the physiopathology of the vascular lesions is well established. Its role in secondary PH is yet to be investigated. We quantified the inflammatory cells on the walls of peripheral pulmonary arteries from patients with congenital heart shunts. METHODS: Twenty-six lung biopsies from patients with increased pulmonary flow and 10 lung fragments from control participants were examined. B-lymphocytes (CD20), T-lymphocytes (CD3), recently recruited macrophages (MAC387) and granulocytes (CD15) were quantified by area of the adventitia in arteries >50 microm. An index of inflammatory cells infiltrating the medial and intimal layers was also determined. RESULTS: There was no difference in the sum of densities of adventitial inflammatory cells between the groups. A prevalence of MAC387-labeled cells was detected in the PH group and of CD3-labeled cells in the controls. There was a lower density of T lymphocytes in the PH group (P<.004). Patients with intimal proliferative lesions showed prevalence of MAC387-labeled cells (P=.004). PH participants showed a higher index of MAC387-labeled cells infiltrating the arterial medial and intimal layers (P<.001). CONCLUSION: The predominance of recently recruited macrophages in the PH group is compatible with ongoing inflammatory reaction in the arterial walls. This could be related to the pathogenesis of the vascular lesions, as a consequence of cytokines produced by the inflammatory cells. The smaller number of adventitial T-lymphocytes in patients with congenital shunts can reflect an impairment of their immune response. PMID- 15358342 TI - Early atherosclerotic lesions of the cardiac conduction system arteries in infants. AB - INTRODUCTION: Although several studies have described initial atherosclerotic lesions of the coronary arteries, already detectable in infancy and even during the intrauterine life, little, if any, attention has been given to the possible involvement of the cardiac conduction system arteries. In particular, to the best of our knowledge, none has considered the lesions of the cardiac conduction arteries as an initial stage of atherosclerosis. METHODS: The cardiac conduction system of 70 infants dying suddenly and unexpectedly was removed in two blocks for paraffin embedding and serially cut. RESULTS: The histological study of the cardiac conduction arteries of the 70 cases examined showed a normal structure in 55 cases (78.57%). In 15 cases (21.43%), there was a thickening of the sinoatrial node and/or atrioventricular artery associated with a thickening of varying severity in coronary artery walls. The lesions were marked by thickening and deposits of amorphous material and mainly lipids in the intima, as well as fragmentation of the elastic fiber system. A significant correlation was evident between early atherosclerotic lesions and both formula feeding and parental cigarette smoking (P<.05, chi(2) test). CONCLUSIONS: The combination of both the considered risk factors seems to increase the early atherogenic effect of each noxa because the coronary lesions were more diffused in formula-fed infants whose parents both smoked. PMID- 15358343 TI - A light-hearted look at a lion-hearted organ (or, a perspective from three standard deviations beyond the norm). Part 1 (of two parts). AB - Throughout history, the heart has been associated not only with its life sustaining function but also with its close ties to the human emotions. In this literature and Internet review, we attempt to gather and organize information from both of these perspectives as they relate to the heart in the following 11 categories: (1) fun facts, (2) medical photography, (3) history, (4) languages (etymology), (5) nonmedical English expressions, (6) death, (7) the arts, (8) movie titles, (9) song titles, (10) Shakespeare, and (11) the Bible. Part 1 will cover the first five topics, and Part 2 will cover the last six topics. These data may be useful to those who are engaged in teaching about the cardiovascular system. PMID- 15358344 TI - Localized atresia of the coronary sinus a short distance proximal to its ostium. AB - Atresia of the ostium of the coronary sinus (CS) is a well-recognized abnormality. We report the incidental finding of atresia of the distal portion of the CS, a short distance proximal to its ostium in a 39-week-old neonate, which, as far as we are aware, has not been previously reported. PMID- 15358345 TI - Cardiac arteriovenous malformation causing sudden death. AB - Cardiac vascular malformations are rare. We report a subendocardial arteriovenous malformation (AVM), associated with extensive myocardial fibrosis, causing sudden death in a 25-year-old woman. To our knowledge, this is the first autopsy case reported. PMID- 15358346 TI - Overexpression of human transforming growth factor-beta1 using a recombinant CHO cell expression system. AB - Transforming growth factor-beta1 (TGF-beta1) is secreted by most cells as a high molecular weight latent complex, which consists of latent TGF-beta1 disulfide bonded to latent TGF-beta1-binding protein (LTBP). Current recombinant expression systems yield less than 1-2 mg of the mature TGF-beta1 per liter of cell culture medium. In an effort to produce large quantities of the recombinant cytokine for structural studies, we have constructed a mammalian expression system based on a modified pcDNA3.1(+) vector with a glutamine synthetase gene inserted for gene amplification. The leader peptide of TGF-beta1 was replaced with that of rat serum albumin, and an eight-histidine tag was inserted immediately after the leader sequence to facilitate protein purification. In addition, Cys 33 of TGF beta1, which forms a disulfide bond with LTBP, was replaced by a serine residue. The resulting expression construct produced a stable clone expressing 30 mg of mature TGF-beta1 per liter of spent medium. Purified TGF-beta1 bound with high affinity to its type II receptor with a solution dissociation constant of approximately 70 nM, and was fully active in both a Mv1Lu cell growth inhibition assay and in a PAI-1 luciferase reporter assay. Owing to similarities in the synthesis, secretion, and structure of TGF-beta family members, this recombinant expression system may also be applied to the overexpression of other TGF-beta isomers and even to members of the TGF-beta superfamily to facilitate their preparation. PMID- 15358347 TI - Expression of tropomyosin from Blattella germanica as a recombinant non-fusion protein in Pichia pastoris and comparison of its IgE reactivity with its native counterpart. AB - Tropomyosins derived from invertebrates are well-known pan allergens. However, the allergenicities of recombinant tropomyosins are variable. Here, we undertook to compare the IgE-binding reactivities of native and recombinant German cockroach tropomyosins. Native tropomyosin was purified by ammonium sulfate fractionation, hydroxyapatite column chromatography, and electroelution, and recombinant tropomyosin was expressed in Pichia pastoris. The allergenicities of the native and recombinant tropomyosins were compared by ELISA inhibition analysis. Native German cockroach tropomyosin showed 18% IgE-binding reactivity to German cockroach sensitized sera. Recombinant tropomyosin was produced without fusion protein and its N-terminus was blocked like that of the native counterpart. The IgE-binding reactivity of the recombinant was found to be comparable to that of native tropomyosin over the concentration range 1-1000 ng/ml by ELISA inhibition testing. Recombinant German cockroach tropomyosin expressed in Pichia pastoris showed better allergenicity than that expressed in Escherichia coli. Other factors in addition to the structural differences of native and recombinant proteins may also influence the IgE reactivities of tropomyosins. PMID- 15358348 TI - Biochemical and immunological characterization of a glycosylated alpha-fucosidase from the invertebrate Unio: interaction of the enzyme with its in vivo binding partners. AB - Mammalian alpha-fucosidase (EC 3.2.1.51) is a lysosomal enzyme that catalyzes the removal of fucose residues from glycosphingolipids and its absence in humans results in a rare metabolic disorder called fucosidosis. Among the invertebrates in the molluscs (Unio) two forms of the enzyme have been reported, a 68 kDa non glycosylated form and a 56 kDa glycosylated form. The glycosylated form has been purified from the seminal fluid of Unio [Biochem. Biophys. Res. Commun. 234 (1997) 54]. In the present study, the 56 kDa glycosylated form has been purified to homogeneity from the whole body tissue of Unio using a series of chromatographic steps. The purified enzyme migrated as a single protein species in 10% SDS-PAGE. Antibodies to the purified enzyme were raised in a rabbit in order to study its biochemical and immunological properties. The purified enzyme is a glycoprotein that exhibits strong binding to Con A-Sepharose gel and can be deglycosylated by PNGase F enzyme suggesting it to be N-glycosylated. The enzyme has been shown to specifically interact with the mannose 6-phosphate receptor protein (MPR 300) purified from goat and Unio. This specific interaction is discussed in view of its possible in vivo binding partners. PMID- 15358349 TI - Purification and characterization of the L-fucose transporter. AB - L-Fucose is a monosaccharide present in low levels in the serum. It is, however, a common structural component of glycoproteins. L-Fucose is accumulated in eukaryotic cells by a specific, facilitative diffusion transport system which has been designated the fucose transporter. In this study, purification of the transporter from mouse brain was performed by detergent extraction followed by ion-exchange and reactive dye ligand column chromatography. Purification was followed using a transport assay into reconstituted liposomes. A 111-fold purification with 5% yield was achieved from the crude homogenate. The apparent molecular weight of the protein was 57 kDa. Transport was found to be saturable. The K(m) and V(max) values are estimated at 3 microM and 275 pmol/min/mg, respectively. The tissue distribution of fucose transport was examined in liver, kidney, heart, lung, spleen, brain, muscle, adipose, ovary, pancreas, and thymus. Some fucose transport was found in all tissues examined. Very low levels were observed in the liver relative to all other tissues examined. The only monosaccharide which could inhibit the uptake of L-[5,6-(3)H]fucose was fucose itself. PMID- 15358350 TI - Function of the signal peptide and N- and C-terminal propeptides in the leucine aminopeptidase from Aeromonas proteolytica. AB - The leucine aminopeptidase from Aeromonas proteolytica (also known as Vibrio proteolyticus) (AAP) is a metalloenzyme with broad substrate specificity. The open reading frame (ORF) for AAP encodes a 54 kDa enzyme, however, the extracellular enzyme has a molecular weight of 43 kDa. This form of AAP is further processed to a mature, thermostable 32 kDa form but the exact nature of this process is unknown. Over-expression of different forms of AAP in Escherichia coli (with AAP's native leader sequence, with and without the N- and/or C terminal propeptides, and as fusion protein) has allowed a model for the processing of wild-type AAP to be proposed. The role of the A. proteolytica signal peptide in protein secretion as well as comparison to other known signal peptides reveals a close resemblance of the A. proteolytica signal peptide to the outer membrane protein (OmpA) signal peptide. Over-expression of the full 54 kDa AAP enzyme provides an enzyme that is significantly less active, due to a cooperative inhibitory interaction between both propeptides. Over-expression of AAP lacking its C-terminal propeptide provided an enzyme with an identical kcat value to wild-type AAP but exhibited a larger Km value, suggesting competitive inhibition of AAP by the N-terminal propeptide (Ki approximately 0.13 nM). The recombinant 32 kDa form of AAP was characterized by kinetic and spectroscopic methods and was shown to be identical to mature, wild-type AAP. Therefore, the ease of purification and processing of rAAP along with the fact that large quantities can be obtained now allow new detailed mechanistic studies to be performed on AAP through site-directed mutagenesis. PMID- 15358351 TI - Expression and purification of a novel rice (Oryza sativa L.) mitochondrial ATP synthase small subunit in Escherichia coli. AB - To clarify the function of the rice mitochondrial ATP synthase 6 kDa subunit (RMtATP6), a method of producing large quantities of this protein is needed. Here, we describe an Escherichia coli expression system for the rapid and economic expression of RMtATP6. The RMtATP6 gene (GenBank Accession No. ) was cloned into the pGEX-6p-3 vector to allow expression of RMtATP6 as a glutathione S-transferase (GST) fusion protein. The RMtATP6-GST fusion protein was purified by affinity chromatography using a glutathione-Sepharose 4B column. A Western blot analysis using anti-GST antibody showed that the fusion protein was not degraded. After enzymatic cleavage of the GST tail, the RMtATP6 protein showed a molecular weight of around 6 kDa. The predicted pI of this protein is 10.01. After improving the conditions of expression and the purification procedures, the final yield of the entire expression and purification process was about 4.6 mg of pure RMtATP6 protein per liter of bacterial culture. PMID- 15358352 TI - Coexpression of yeast copper chaperone (yCCS) and CuZn-superoxide dismutases in Escherichia coli yields protein with high copper contents. AB - To fully understand the function of the Cu- and Zn-containing superoxide dismutases in normal and disordered cells, it is essential to study protein variants with full metal contents. We describe the use of an Escherichia coli based expression system for the overproduction of human intracellular wild type CuZn-superoxide dismutase (SOD), the CuZnSOD variant F50E/G51E (monomeric), two amyotrophic lateral sclerosis-related mutant CuZnSOD variants (D90A and G93A), and PseudoEC-SOD, all with high Cu contents. This system is based on coexpression of the SOD variants with the yeast copper chaperone yCCS during growth in a medium supplemented with Cu(2+) and Zn(2+). The recombinant SOD enzymes were all found in the cytosol and represented 30-50% of the total bacterial protein. The enzymes were purified to homogeneity and active enzymes were obtained in high yield. The resulting proteins were characterized through immunochemical reactivity and specific activity analyses, in conjunction with mass-, photo-, and atomic absorption-spectroscopy. PMID- 15358353 TI - Cloning, expression, and purification of the virulence-associated protein D from Xylella fastidiosa. AB - In this study, an efficient expression system, based on the pET32Xa/LIC vector, for producing a Xylella fastidiosa virulence-associated protein D, found to have a strong similarity to Riemerella anatipestifer and Actinobacillus actinomycetencomitans VapD protein, is presented. The protein has a molecular mass of 17.637 Da and a calculated pI of 5.49. The selected XFa0052 gene was cloned in the pET32Xa/LIC vector and the plasmid was transformed into Escherichia coli BL21 (DE3) strain at 37 degrees C, with an induction time of 2 h and 1 mM IPTG concentration. The protein present in the soluble fraction was purified by immobilized metal affinity chromatography (IMAC), and had its identity determined by mass spectrometry (MALDI-TOF) and N-terminal sequencing. The purified protein was found as a single band on SDS-PAGE and its correct folding was verified by circular dichroism spectroscopy. PMID- 15358354 TI - Regulated expression of sFRP-1 protein by the GeneSwitch system. AB - The GeneSwitch system is a mifepristone-inducible expression system that provides exceptionally low uninduced and high-induced protein expression in mammalian cells. We have developed an adenovirus recombinant containing GeneSwitch protein driven by the GAL4-tk promoter, as well as recombinants containing sFRP-1 and luciferase reporter under the control of the GAL4-E1b promoter. Luciferase activity in A549 cells infected with the GeneSwitch and Luciferase viruses is very low in ethanol-treated cells, while the level of luciferase activity increases 200-fold in cells treated with mifepristone. Conditional expression of functional sFRP-1 is demonstrated in A549, human osteoblast, and CHO cell lines by either the co-infection of cells with sFRP-1 and GeneSwitch viruses or the infection of GeneSwitch expressing cell lines with sFRP-1 virus and subsequent treatment with mifepristone. The expression of sFRP-1 is seen as early as 4 h post-mifepristone treatment, reaching the highest levels at 20 h. The sFRP-1 protein is present in conditioned media, and the protein is functional based upon its ability to inhibit the Wnt-mediated activation of TCF-Luciferase reporter activity. The regulated expression of sFRP-1 utilizing adenovirus vectors provides an opportunity to address the contribution of sFRP-1 in the regulation of stem cell differentiation, maturation, and their function by modulating the Wnt signaling. PMID- 15358355 TI - Recombinant expression of Ole e 6, a Cys-enriched pollen allergen, in Pichia pastoris yeast: detection of partial oxidation of methionine by NMR. AB - Olive pollen is one of the main causes of allergy in Mediterranean countries. Ole e 6, an olive pollen allergen, is a small (5.8 kDa) and acidic protein (pI 4.2) and no homologous proteins have been isolated or characterized so far. Ole e 6 has been efficiently expressed in the methylotrophic yeast Pichia pastoris. The cDNA encoding Ole e 6 was inserted into the plasmid vector pPIC9 and overexpressed in GS115 yeast cells. The recombinant product was purified by size exclusion chromatography followed by reverse-phase HPLC. N-terminal sequencing, amino acid composition analysis, CD, NMR, and IgG-binding experiments were employed to characterize the purified protein. NMR data revealed the oxidation of the methionine at position 28 in approximately 50% of the recombinant protein but, although this alters its electrophoretic behavior, it did not affect folding or IgG-binding properties of rOle e 6. The recombinant form of Ole e 6 expressed in P. pastoris can be employed for structural and biochemical studies. PMID- 15358356 TI - Expression and purification of His-tagged rat mitochondrial short-chain 3 hydroxyacyl-CoA dehydrogenase wild-type and Ser137 mutant proteins. AB - Mitochondrial 3-hydroxyacyl-CoA dehydrogenase is a key enzyme in the beta oxidation of fatty acids. The deficiency of this enzyme in patients has been previously reported. We cloned the gene of rat mitochondrial 3-hydroxyacyl-CoA dehydrogenase in a bacterial expression vector pLM1 with six continuous histidine codons attached to the 5' of the gene. The cloned gene was overexpressed in Escherichia coli and the soluble protein was purified with a nickel HiTrap chelating metal affinity column to apparent homogeneity. The specific activity of the purified His-tagged rat mitochondrial 3-hydroxyacyl-CoA dehydrogenase was 452 U/mg. Ser137 is a highly conserved amino acid, which, it has been suggested, is an important residue because of its proximity to the modeled L-3-hydroxyacyl-CoA substrate in the crystal structure of 3-hydroxyacyl-CoA dehydrogenase. We constructed three mutant expression plasmids of the enzyme using site-directed mutagenesis. Mutant proteins were overexpressed in E. coli and purified with a nickel metal affinity column. Kinetic studies of wild-type and mutant proteins were carried out, and the result confirmed that Ser137 is a very important residue of rat mitochondrial 3-hydroxyacyl-CoA dehydrogenase. Our overexpression in E. coli and one-step purification of the highly active rat mitochondrial 3 hydroxyacyl-CoA dehydrogenase greatly facilitated our further investigation of this enzyme, and our result from site-directed mutagenesis increased our understanding of 3-hydroxyacyl-CoA dehydrogenase. PMID- 15358357 TI - A spectrophotometric assay of D-glucuronate based on Escherichia coli uronate isomerase and mannonate dehydrogenase. AB - Escherichia coli uronate isomerase and mannonate dehydrogenase were overexpressed in E. coli BL21(DE3)pLysS cells and purified to near-homogeneity. The kinetic properties of the two enzymes were investigated. The isomerase was found to be inhibited by EDTA and to be stimulated by Zn(2+), Co(2+), and Mn(2+), but not by Mg(2+) or Ca(2+). Both enzymes were used to develop a sensitive spectrophotometric assay, in which D-glucuronate is converted to D-mannonate with concomitant oxidation of NADH to NAD(+). The sensitivity of this assay permits the detection of less than 1 nmol D-glucuronate. This assay can also be used to determine the concentration of beta-glucuronides and glucuronate 1-phosphate after enzymatic hydrolysis of these compounds with beta-glucuronidase or alkaline phosphatase. PMID- 15358358 TI - Intein-mediated fusion expression, high efficient refolding, and one-step purification of gelonin toxin. AB - An open reading frame of gelonin (Gel), one of ribosome inactivating proteins, was inserted into the vector pBSL-C which contains the coding region of chitin binding domain (CBD)-intein, resulting in the fusion expression of CBD-intein-Gel in Escherichia coli BL21 (DE3) by the induction of IPTG. The fusion product formed an aggregate of the misfolded protein, commonly referred to as inclusion bodies (IBs). The IBs were denatured and then refolded by step-wise dialysis. About 69% fusion protein was in vitro refolded to native state in the presence of GSSG and GSH as monitored by size-exclusion HPLC. The refolded CBD-intein-Gel was loaded onto chitin beads column equilibrated with 10 mM Tris buffer, 500 mM NaCl, pH 8.5, and about 2.4 mgGel/L culture with 96% homogeneity was directly eluted from the captured column by incubation at 25 degrees C under pH 6.5 for 48 h based on intein C-terminal self-cleavage. Western blot, ELISA, and in vitro inhibition of protein synthesis demonstrated that the bioactivity of recombinant Gel was comparable to that of native Gel purified from seeds. This implied that the purified Gel by this method is biologically active and suitable for further studies. PMID- 15358359 TI - Expression and characterization of isoform 1 of human mitochondrial elongation factor G. AB - Elongation factor G (EF-G) catalyzes the translocation step of protein biosynthesis. Genomic analysis suggests that two isoforms of this protein occur in mitochondria. The region of the cDNA coding for the mature sequence of isoform 1 of human mitochondrial EF-G (EF-G1(mt)) has been cloned and expressed in Escherichia coli. The recombinant protein has been purified to near homogeneity by chromatography on Ni-NTA resins and cation exchange high performance liquid chromatography. EF-G1(mt) is active on both bacterial and mitochondrial ribosomes. Human EF-G1(mt) is considerably more resistant to fusidic acid than many bacterial translocases. A molecular model for EF-G1(mt) has been created and analyzed in the context of its relationship to the translocases from other systems. PMID- 15358360 TI - Improved PC1/3 production through recombinant expression in insect cells and larvae. AB - Protein convertase 1/3 is a serine endoproteinase present in the regulated secretory pathway of endocrine and neuroendocrine cells. It is responsible for the processing of numerous prohormones and proneuropeptides into their biologically active moieties, often following cleavage at pairs of basic residues. The determination of its three-dimensional structure, as well as the understanding of its enzymatic properties, would greatly benefit from the production and availability of large amounts of recombinant enzyme. We report herein improvements in the production of PC1/3 by expressing recombinant mutated forms in both insect cells (Spodoptera frugiperda, Sf9 cells) and larvae (Trichoplusia ni commonly referred to as cabbage looper). On one hand, we deleted the last 135 COOH-terminal residues of mPC1/3 and, on the other hand, we replaced the signal peptide of mPC1/3 by the viral glycoprotein gp67 signal peptide. These modifications were shown to improve markedly (up to 125%) the secretion into the Sf9 cells medium and the amount of enzymatic activity recovered when compared to the original vector. Moreover, intracoelemic injection of the vectors into insect larvae led to the production and purification of enzymatically active enzyme at a level of 30 microg/larva in the case of mPC1/3 and to the production of a high amount of another enzymatically active convertase, PC7. The optimal viral titer for infection of larvae was determined to be 10(6)pfu/ml. Taking into account the purification protocol combined with the ease and efficiency of using larvae, it should now be possible to meet the needs for biochemical and structural studies. PMID- 15358361 TI - Expression and purification of thioredoxin (TrxA) and thioredoxin reductase (TrxB) from Brevibacillus choshinensis. AB - Brevibacillus choshinensis (formerly Bacillus brevis) is a protein-hyperproducing bacterium and has been used for commercial protein production. Here, we cloned thioredoxin (trxA) and thioredoxin reductase (trxB) genes from B. choshinensis, and expressed the gene products in Escherichia coli with an amino-terminal hexa His-tag for purification and characterization. His-TrxA and His-TrxB were purified to homogeneity with one-step Ni-NTA affinity column chromatography, and the two recombinant proteins showed identical specific activity with or without removal of the amino-terminal His-tag, indicating that the extrasequence containing the hexa-His-tag did not affect their enzymatic activities. The E. coli expression system used here resulted in a 40-fold increase in production of His-TrxB protein compared to the level of native TrxB produced in non-recombinant B. choshinensis cells. TrxA and TrxB proteins with carboxy-terminal His-tag (TrxA His and TrxB-His) were successfully expressed in B. choshinensis and were purified by Ni-NTA column chromatography. Co-expression of TrxA-His with recombinant human epidermal growth factor (hEGF) in B. choshinensis promoted the extracellular production of hEGF by up to about 200%. PMID- 15358362 TI - Optimised expression and purification of recombinant human indoleamine 2,3 dioxygenase. AB - The hemoprotein indoleamine 2,3-dioxygenase (IDO) is the first and rate-limiting enzyme in mammalian tryptophan metabolism. It has received considerable attention in recent years, particularly due to its role in the pathogenesis of many diseases. Here, we report attempts to improve soluble expression and purification of hexahistidyl-tagged recombinant human IDO from Escherichia coli (EC538, pREP4, and pQE9-IDO). Significant formation of inclusion bodies was noted at the growth temperature of 37 degrees C, with reduced formation at 30 degrees C. The addition of the natural biosynthetic precursor of protoporphrin IX, delta-aminolevulinic acid (ALA), coupled with optimisation of IPTG induction levels during expression at 30 degrees C and purification by nickel-agarose and size exclusion chromatography, resulted in protein with 1 mol of heme/mol of protein and a specific activity of 160 micromol of kynurenine/h/mg of protein (both identical to native human IDO). The protein was homogeneous in terms of electrophoretic analysis. Yields of soluble protein (3-5 mg/L of bacterial culture) and heme content are greater than previously reported. PMID- 15358363 TI - Production and purification of a chimeric monoclonal antibody against botulinum neurotoxin serotype A. AB - Production of recombinant antibodies against botulinum neurotoxin is necessary for the development of a post-exposure treatment. CHO-DG44 cells were transfected with a plasmid encoding the light and heavy chains of a chimeric monoclonal antibody (S25) against botulism neurotoxin serotype A. Stable cell lines were obtained by dilution cloning and clones were shown to produce nearly equivalent levels of light and heavy chain antibody by an enzyme-linked immunosorbent assay (ELISA). In suspension culture, cells produced 35 microg/ml of chimeric antibody after 6 days, corresponding to a specific antibody productivity of 3.1 pg/cell/day. A method for the harvest and recovery of an antibody against botulism neurotoxin serotype A was investigated utilizing ethylenediamine-N,N' tetra(methylphosphonic) acid (EDTPA) modified zirconia and MEP-hypercel, a hydrophobic charge interaction chromatography resin. Purification of the S25 antibody was compared to that achieved using rProtein A-Sepharose Fast Flow resin. After the direct load of culture supernatant, analysis by ELISA and gel electrophoresis showed that S25 antibody could be recovered at purities of 41 and 44%, from the EDTPA modified zirconia and MEP-hypercel columns, respectively. Although the purity obtained from each of these columns was low, the ability to withstand high column pressures and nearly 90% recovery of the antibody makes EDTPA modified zirconia well suited as an initial capture step. Combining the EDTPA modified zirconia and HCIC columns in series resulted in both purity and final product yield of 72%. PMID- 15358364 TI - Functional transplantation of the sumoylation machinery into Escherichia coli. AB - Modification by SUMO proteins appears to be very common in eukaryotic cells. Many proteins have been reported to be sumoylated, at least under certain circumstances, in vivo, and new examples get published every month. On the other hand, sumoylation is, in essence, a way to construct branched proteins or protein fusions. Obtention of pure sumoylated proteins from eukaryotic cells is not easy because of the dynamic nature of this modification and the large number of sumoylated proteins in vivo. Production of sumoylated proteins in vitro requires the previous purification of most of the components of the pathway, and has the typical limitations of such systems. In this paper, we describe a method to quantitatively produce sumoylated proteins in vivo in Escherichia coli as a way to obtain large quantities of specifically sumoylated target proteins with a high degree of purity, to generate fusion proteins not limited to N- or C-end additions, and to polymerize proteins by covalent linkage. PMID- 15358365 TI - Refolding of the Cupressus arizonica major pollen allergen Cup a1.02 overexpressed in Escherichia coli. AB - The cDNA encoding an isoform of the cypress major pollen allergen, Cup a1.02, has been cloned and expressed in Escherichia coli as a N-terminal 6x His-tagged protein. To increase recovery, Cup a1.02 was expressed at high levels exploiting the T5 strong promoter and led to accumulate as inclusion bodies. The insoluble purified aggregates were solubilized in 6 M guanidine hydrochloride, immobilized using nickel-chelating affinity chromatography, and successfully refolded by controlled removal of the chaotropic reagent. Enhanced protein refolding was observed by reducing the protein concentration at 0.6-0.8 mg/ml. SDS-PAGE and gel filtration chromatography indicated an apparent molecular mass of approximately 40 kDa and the occurrence of the protein as monomers. The reconstituted fusion protein displayed the same immunological properties of the native Cup a1.02 protein as proven by IgE immunoreactivity. Immunoblotting, ELISA, and histamine release test showed that the tag did not preclude the protein functionality hence validating its correct three-dimensional folding. The protein fold was also assessed by CD spectroscopy and deconvolution of the spectrum allowed to estimate the secondary structure as a prevalence of beta structures (higher than 60%) and a small contribution from alpha helices (less than 12%). The reported procedure has proven to be useful for the production of multi-milligrams of recombinant Cup a1.02 allergen suitable for structural biology studies and for the molecular and functional characterization of the IgE binding sites. PMID- 15358366 TI - Expression of enterovirus 70 capsid protein VP1 in Escherichia coli. AB - The VP1 gene of enterovirus 70 (EV70) possesses a large number of Escherichia coli low-usage codons (11.0%) and a bacterial ribosome binding site complementary sequence (RBSCS) 5'-UGUCUCCUUUUC-3' flanking the codon 139. Plasmids containing EV70 cDNA encoding the full-length VP1 failed to express in E. coli (BL21(DE3), Rosetta 2(DE3) or Rosetta (DE3)pLysS). High expression (>8% of total protein) of recombinant VP1 (rVP1m) in E. coli required engineering of the encoding cDNA (conserved modification of the native cDNA) by simultaneous substitution of a rare-codon cluster located between codons 103 and 132, and replacement of the RBSCS-TCCTTT sequence. The rare-codon frequencies of the cDNAs encoding VP1 non overlapping terminal fragments N138 (1-138 aa) and C170 (141-310 aa) are similar (10.9 and 11.2%, respectively). However, in E. coli, high expression of recombinant C170 (rC170) required no modification of the native cDNA whereas high expression of recombinant N138 (rN138m) required minimal synonymous substitution of the above rare-codon cluster. The rare-codon cluster of EV70 VP1 gene has five least-usage arginine codons (AGG/AGA) and three tandem rare-codon pairs (AGGAGG, CUAAGG, and AGACUA). Our results suggest that the rare-codon cluster (its rare codon arrangement per se and/or its related mRNA secondary structure(s)) and the RBSCS in EV70 VP1 gene, not the rare-codon frequency, constitute the key elements that suppress its expression in E. coli. PMID- 15358367 TI - Over-expression in Escherichia coli, functional characterization and refolding of rat dimethylglycine dehydrogenase. AB - Dimethylglycine dehydrogenase (Me(2)GlyDH) is a mitochondrial enzyme that catalyzes the oxidative demethylation of dimethylglycine to sarcosine. The enzyme requires flavin adenine dinucleotide (FAD), which is covalently bound to the apoprotein via a histidyl(N3)-(8alpha)FAD linkage. In the present study, the mature form of rat Me(2)GlyDH has been over-expressed in Escherichia coli as an N terminally 6-His-tagged fusion protein. The over-expressed protein distributed almost equally between the soluble and insoluble (inclusion bodies) cell fraction. By applying the soluble cell lysate to a nickel-chelating column, two fractions were eluted, both containing a nearly homogeneous protein with a molecular mass of 93 kDa, on SDS-PAGE. The first protein fraction was identified by Western blotting analysis as the covalently flavinylated Me(2)GlyDH. It showed optical properties and specific activity (240 nmol/min/mg protein) similar to those of the native holoenzyme. The second fraction was identified as an underflavinylated (apo-) form of Me(2)GlyDH, with a 70% lower specific activity. The recombinant holoenzyme exhibited optimal activity at pH 8.5, an activation energy of about 80 kJ/mol, and two KM values for N,N-dimethylglycine (KM1 = 0.05 mM and KM2 = 9.4 mM), as described for the native holoenzyme. Starting from the inclusion bodies, the unfolded flavinylated enzyme was solubilized by SDS treatment and refolded by an 80-fold dilution step, with a reactivation yield of 50-60%. PMID- 15358368 TI - Expression, purification, and initial structural characterization of rat orphan nuclear receptor NOR-1 LBD domain. AB - NOR-1 is an orphan member of the nuclear receptor superfamily, which includes a group of transcription factors involved in the response to steroids, fatty acids, retinoic acids, and other lipophilic molecules. The NOR-1 subfamily (NR4), composed also of Nurr1 and Nurr77, has been implicated in cell proliferation, differentiation, apoptosis, chondrosarcomas, inflammation, and atherogenesis. The NOR-1 receptor is an orphan ligand receptor which acts over gene transactivation. No ligands, if such in fact exist, are known for this receptor. Recently, the three-dimensional structure of the homolog receptor Nurr1 has been solved using protein crystallography techniques. Surprisingly, the structure does not present either a typical cavity for ligand binding or a classical co-factor binding site in the ligand binding domain (LBD). To allow for structural studies of other members of NR4 subfamily, we have subcloned, overexpressed in Escherichia coli cells, purified, and characterized the rat orphan nuclear receptor NOR-1 LBD domain. We obtained NOR-1 LBD at a high degree of purity and with an overall yield of 3 mg/L of culture media. CD spectroscopic analysis shows a high alpha helical secondary structure content (52%), similar to that of Nurr 1 LBD three dimensional structure. Thermal denaturation monitored by UV absorption and CD spectroscopy suggests proper folding of recombinant NOR-1 LBD. PMID- 15358369 TI - Purification of recombinant human carbonic anhydrase-II by metal affinity chromatography without incorporating histidine tags. AB - Due to their involvement in diverse pathological conditions, carbonic anhydrases have been the targets of drug developments for the treatments of glaucoma, epilepsy, high altitude sickness, as well as cancer. Of about 14 isozymes of carbonic anhydrases, carbonic anhydrase-II (hCA-II) has been most extensively investigated from the structural, functional, and inhibitor design point of view. We discovered that hCA-II preferentially binds to the Sepharose-iminodiacetate (IDA)-Zn(2+) column, and such binding does not require incorporation of either N- or C-terminal histidine tags in the protein structure. By using the Sepharose-IDA Zn(2+) affinity column, we purified the Escherichia coli expressed hCA-II with an overall recovery of 76%. The purified enzyme showed a single band on the SDS PAGE. Due to ease in preparing the Sepharose-IDA-Zn(2+) column, and purifying hCA II just in one step, the overall protocol will be ideal for producing bulk quantities of the enzyme for high throughput screening of inhibitors. PMID- 15358370 TI - Overexpression and functional characterisation of the human melanocortin 4 receptor in Sf9 cells. AB - The human melanocortin 4 receptor (MC4r) was successfully expressed in Sf9 cells using the baculovirus infection system. N- and C-terminally His-tagged receptors generated B(max) values of 14 and 23 pmol receptor/mg membrane protein, respectively. The highest expression level obtained with the C-terminally His tagged MC4r corresponded to 0.25mg active receptor/litre culture volume. Addition of a viral signal peptide at the N-terminus of the His-tagged MC4r did not improve the expression level. Confocal laser microscopy studies revealed that both the N- and C-terminally tagged MC4r did not accumulate intracellularly and were mainly located in the plasma membrane. The recombinant receptors showed similar affinity for the agonist NDP-MSH (Kd = 11 nM) as to MC4r expressed in mammalian cells. Functional coupling of the highest expressed C-terminal tagged receptor to endogenous Galpha protein was demonstrated through GTPgammaS binding upon agonist stimulation of the receptor. Ki values for the ligands MTII, HS014, alpha-, beta-, and gamma-MSH are comparable to the values obtained for MC4r expressed in mammalian cells. PMID- 15358371 TI - Haemophilus influenzae UvrA: overexpression, purification, and in cell complementation. AB - UvrA protein is a major component of ABC endonuclease complex involved in nucleotide excision repair (NER) mechanism. Although NER system is best characterized in Escherichia coli, not much information is available in Haemophilus influenzae. However, based on amino acid homology, uvrA ORF has been identified on H. influenzae genome [gene identification No. HI0249, Science 269 (1995) 496]. H. influenzae Rd uvrA ORF was cloned and overexpressed in E. coli. The expressed UvrA protein was purified using a two-step column chromatography protocol to a single band of expected molecular weight (104 kDa) and characterized for its ATPase and DNA binding activity. In addition, when H. influenzae uvrA was introduced in E. coli uvrA mutant strain AB1886, its UV resistance was restored to near wild type level. PMID- 15358372 TI - Simple separation of DNA in antibody purification. AB - Producing monoclonal antibodies includes their efficient and simple purification. Growing hybridoma cells in media containing Prolifix, an alternative plant-based substitute for serum, provides supernatants containing large amounts of antibodies and defined low molecular weight additives. Antibodies can easily be separated from these compounds by fast ultrafiltration. However, DNA originating from lysed cells is present in substantial amounts and must be removed for most antibody applications. The present communication provides a fast, cheap, and efficient separation method by precipitating the DNA from a phosphate buffered solution with manganese chloride. Resulting antibodies have a high purity and an unchanged bioactivity. The method is especially valuable for antibodies which lose bioactivity by interactions with chromatographic matrices (as, for example, Sepharose) and can be used for various antibody isotypes. PMID- 15358373 TI - Expression and purification of His-tagged rat mitochondrial medium-chain acyl-CoA dehydrogenase wild-type and Arg256 mutant proteins. AB - Mitochondrial medium-chain acyl-CoA dehydrogenase is a key enzyme for the beta oxidation of fatty acids, and the deficiency of this enzyme in patient has been previously reported. We cloned the gene of rat mitochondrial medium-chain acyl CoA dehydrogenase into a bacterial expression vector pLM1 with six continuous histidine codons attached to the 3' of the gene. The cloned gene was overexpressed in Escherichia coli and the soluble protein was purified with a nickel Hi-Trap chelating metal affinity column in 88% yield to apparent homogeneity. The specific activity of the purified His-tagged rat mitochondrial medium-chain acyl-CoA dehydrogenase was 4.0 U/mg. Arg256 is a highly conserved amino acid, which may play an important role in enzymatic reaction based on the crystal structure of medium-chain acyl-CoA dehydrogenase. We constructed four mutant expression plasmids of the enzyme using site-directed mutagenesis. Mutant proteins were overexpressed in E. coli and purified with a nickel metal affinity column. Kinetic studies of wild-type and mutant proteins were carried out, and the result confirmed that Arg256 is a very important residue of rat mitochondrial medium-chain acyl-CoA dehydrogenase. Our overexpression in E. coli and one-step purification of the highly active rat mitochondrial medium-chain acyl-CoA dehydrogenase greatly facilitated our further investigation of this enzyme, and our result from site-directed mutagenesis increased our understanding of medium chain acyl-CoA dehydrogenase. PMID- 15358374 TI - Ni-chelate-affinity purification and crystallization of the yeast mitochondrial F1-ATPase. AB - The yeast mitochondrial ATPase has been genetically modified to include a His(6) Ni-affinity tag on the amino end of the mature beta-subunit. The modified beta subunit is imported into the mitochondrion, properly processed to the mature form, and assembled into a mature and fully active ATP synthase. The F(1)-ATPase has been purified from submitochondrial particles after release from the membrane with chloroform, followed by Ni-chelate-affinity and gel filtration chromatography. The final enzyme is a homogeneous preparation with full activity and no apparent degradation products. This enzyme preparation has been used to obtain crystals that diffract to better than 2.8 A resolution. PMID- 15358375 TI - Expression and characterization of extracellular fungal phytase in transformed sesame hairy root cultures. AB - A recombinant fungal phytase was produced by cultures of sesame hairy roots transformed with Agrobacterium rhizogenes, purified and its molecular properties were characterized. Its transcription level and the phytase production were rapidly increased after 4 weeks of the cultures, suggesting that its transcription and protein synthesis might concur. Western blot analysis provided evidence that the recombinant fungal phytase was secreted into the liquid culture medium of the hairy roots. The phytase enzyme secreted was purified by three steps of ultrafiltration, DEAE-Sepharose ion exchange chromatography, and Sephadex G-100 size-exclusion chromatography. As a result, one single band signal was observed with SDS-PAGE, indicating that the purification step was reasonable. The positive signs of both the zymogram and the PAS staining on SDS-PAGE suggested that the activity of the final product phytase was active and glycosylated. The optimal reaction temperature of the phytase was between 50 and 60 degrees C and at over 60 degrees C its activity was reduced by 30-90%, depending on the temperatures applied. Pre-incubation at temperatures of 20-50 degrees C showed stable catalytic activity, while at over 50 degrees C the phytase activity was gradually decreased by 90%. The optimal pH was between 4 and 5 pH values for the recombinant fungal phytase, while for native phytase it was at pH 5.0. Addition of iron ion inhibited the phytase activity but treatments of some cations, EDTA, and PMSF showed no effect on the activity or slightly stimulated it positively. PMID- 15358376 TI - Over-expression of the human MDM2 p53 binding domain by fusion to a p53 transactivation peptide. AB - MDM2 binds to the tumor suppressor protein p53 and regulates the level of p53 in cells. Although it is possible to prepare a small amount of the region of MDM2 that binds to p53, the expression level of this fragment of MDM2 is relatively low, limiting the studies involving this protein. Here, we describe a construct for the optimized bacterial expression and purification of the MDM2 p53 binding domain. We found that the expression level of the soluble MDM2 p53 binding domain in bacteria was increased dramatically by fusing it to its interaction partner, the p53 transactivation peptide. Attachment of the p53 transactivation peptide (residues 17-29) to the N-terminus of MDM2 resulted in a more than 200-fold increase of soluble protein expression of the p53 binding domain in bacteria. To obtain the final MDM2 p53 binding domain (residues 5-109) we inserted a tobacco etch virus protease recognition site between the P53 peptide and the MDM2 p53 binding domain. To weaken the protein/peptide interaction and facilitate the separation of the protein from the complex, we introduced a point mutation of one of the key interaction residues (F19A or W23A) in the p53 peptide. The advantages of our new construct are high yield and easy purification of the MDM2 protein. PMID- 15358377 TI - Efficient and rapid purification of recombinant human alpha-galactosidase A by affinity column chromatography. AB - The lysosomal enzyme alpha-galactosidase A (alpha-Gal A) metabolizes neutral glycosphingolipids that possess alpha-galactoside residues at the non-reducing terminus, and inherited defects in the activity of alpha-Gal A lead to Fabry disease. We describe here an efficient and rapid purification procedure for recombinant alpha-Gal A by sequential Concanavalin A (Con A)-Sepharose and immobilized thio-alpha-galactoside (thio-Gal) agarose column chromatography. Optimal elution conditions for both columns were obtained using overexpressed human alpha-Gal A. We recommend the use of a mixture of 0.9 M methyl alpha mannoside and 0.9 M methyl alpha-glucoside in 0.1 M acetate buffer (pH 6.0) with 0.1 M NaCl for the maximum recovery of glycoproteins with multiple high-mannose type sugar chains from Con A column chromatography, and that the Con A column should not be reused for the purification of glycoproteins that are used for structural studies. Binding of the enzyme to the thio-Gal column requires acidic condition at pH 4.8. A galactose-containing buffer (25 mM citrate-phosphate buffer, pH 5.5, with 0.1 M galactose, and 0.1 M NaCl) was used to elute alpha-Gal A. This procedure is especially useful for the purification of mutant forms of alpha-Gal A, which are not stable under conventional purification techniques. A protocol that purifies an intracellular mutant alpha-Gal A (M279I) expressed in COS-7 cells within 6h at 62% overall yield is presented. PMID- 15358378 TI - Management of acute severe asthma. AB - Asthma is a chronic inflammatory disorder that results in recurrent episodes of reversible airflow obstruction. Lung hyperinflation results from obstruction or dynamic airway collapse during exhalation. Obstruction and dynamic hyperinflation both play a deleterious role in asthma. Patients who present with asthma have increased inspiratory work of breathing due to lung hyperinflation and auto positive end-expiratory pressure (auto-PEEP). The goal of acute care treatment is to reverse bronchoconstriction and inflammation, thus reducing dynamic hyperinflation, so that breathing is restored to baseline, unlabored, quiet breathing. PMID- 15358379 TI - Pharmacologic therapy for acute exacerbations of chronic obstructive pulmonary disease: a review. AB - This article reviews available data on the drug therapy armamentarium for the acute exacerbation of chronic obstructive pulmonary disease (COPD). Summaries of studies and therapeutic issues for bronchodilators, antibiotic therapy, corticosteroid use, and a few miscellaneous agents are presented. Many controversies exist in the criteria defining the acute exacerbation, in defining appropriate outcome parameters for assessment, and, consequently, in developing specific consistent recommendations for drug therapy. Five published guidelines assist the clinician in therapeutic drug management of the acute exacerbation of COPD, and each differs in its recommendations for drug therapy prescription. The article includes synopses for drug therapy recommendations from the guidelines. PMID- 15358380 TI - Caring for the bariatric patient with obstructive sleep apnea. AB - Caring for the bariatric patient in the acute care environment poses many challenges to the team delivering care. The management of obstructive sleep apnea with noninvasive ventilation is one priority. Treatment options, including bi level ventilation and continuous positive airway pressure, and delivery strategies are reviewed in conjunction with other aspects of care that ensure a holistic, comprehensive plan of care. The most effective plan of care for the bariatric patient is one that uses a multidisciplinary coordinated team effort. PMID- 15358381 TI - Prone positioning in the patient who has acute respiratory distress syndrome: the art and science. AB - Acute respiratory distress syndrome (ARDS) remains a significant contributor to the morbidity and mortality of patients in the ICU. A variety of treatments are used to support the lung of the patient who has ARDS and improve gas exchange during the acute injury phase. It seems, however, that the simple, safe, and noninvasive act of prone positioning of the critically ill patient who has ARDS may improve gas exchange while preventing potential complications of high positive end-expiratory pressure, volutrauma, and oxygen toxicity. This article provides the critical care nurse with the physiologic rationale for use of the prone position, indications and contraindications for use, safe strategies for prone positioning, and care techniques and monitoring methods of the patient who is in the prone position. PMID- 15358382 TI - Airway pressure release ventilation and pediatrics: theory and practice. AB - Airway pressure release ventilation (APRV) facilitates oxygenation and ventilation by maintaining an elevated baseline airway pressure similar to continuous positive airway pressure (CPAP). APRV differs from CPAP only by the addition of regular, brief release of airway pressure to facilitate carbon dioxide removal. The baseline pressure maintains a near continuous airway pressure to facilitate recruitment, improving ventilation and oxygenation. Pediatric patients who have recruitable lung disease may be well suited to the application of APRV. PMID- 15358383 TI - Preventing ventilator-associated pneumonia: evidence-based care. AB - Ventilator-associated pneumonia is a common complication of mechanical ventilation with significant morbidity and mortality. This article summarizes the data related to specific risk factors associated with ventilator associated pneumonia (patient position, oral health, airway management, and gastrointestinal factors) and provides recommendations for practice based on the present evidence. PMID- 15358384 TI - Sedation assessment: easier said than done! AB - Sedation assessment commonly is performed in critically ill patients to evaluate their need for, and response to, sedation therapy. Although many sedation assessment scales have been published, few have been tested properly to assure their appropriateness for use in critically ill patients. This article highlights the published sedation assessment scales that have the strongest scientific basis, identifies limitations of the current scales, and suggests characteristics for future sedation assessment scales that would overcome many of the current problems. PMID- 15358385 TI - Sedation and comfort issues in the ventilated infant and child. AB - Intubated infants and children require optimal sedation and comfort measures. Key elements that ensure the provision of quality care for these patients include the use of pain, comfort, and sedation assessment tools; pharmacologic and nonpharmacologic strategies; and the inclusion of the patient and parents as part of the team. This article describes approaches for using sedation and ensuring comfort in these patients. Application of research and the education of team members and the patient and family are crucial aspects of care and are also discussed. PMID- 15358386 TI - The science of weaning: when and how? AB - Weaning patients from long-term mechanical ventilation continues to be a goal of clinicians and scientists and the hospitals charged with their care. This article describes the science of the "wean" and the "how" of weaning. A goal of scientists has been to develop predictors that determine accurately the optimal time to initiate weaning. Unfortunately to date none has emerged as superior. Quite simply, predictors do not predict. In contrast, methods that decrease variation in care practices have demonstrated positive outcomes. The methods include protocols for weaning trials and sedation and other system initiatives inclusive of a multidisciplinary plan of care or clinical pathway. PMID- 15358387 TI - Weaning of the neurologically impaired patient. AB - Many studies have been published on weaning patients from mechanical ventilation, but few have addressed the unique needs of patients with neurologic impairment. Typically, neuroscience patients remain intubated because of concerns over airway protection or neuromuscular weakness. This article discusses special weaning considerations for this patient population. Neurologic-specific weaning trends from a comprehensive ventilator weaning program are also presented. PMID- 15358388 TI - The experience of four outcomes managers: an institutional approach to weaning patients from long-term mechanical ventilation. AB - Care of patients requiring long-term mechanical ventilation (LTMV) is the focus of many hospitals nationwide. Because the care of patients who require LTMV is complex and morbidity and mortality are high, associated costs often exceed reimbursement. This article describes a successful institutional program for the care of the patient population requiring LTMV using advanced practice nurses (APNs) in the role of outcomes managers (OMs). So that others may benefit from the experiences of the clinicians, this article describes the historical background, the selection and qualifications of APNs, the process APNs use to manage and monitor the patients, role challenges, and associated outcomes. PMID- 15358389 TI - Contemporary issues in adult tracheostomy management. AB - Airway assessment and management is one of the primary responsibilities of nurses caring for the critically ill adult. Common types of artificial airways, such as endotracheal and tracheostomy tubes, require clinicians to have a complete understanding of indications, complications, and clinical applications. Although tracheostomy tubes are used to a lesser extent than endotracheal tubes in most ICUs, their use typically requires additional education and training because of the complexity and diversity of available tubes, care of surgical site, and other related nursing care issues. The purpose of this article is to provide a review of current practice trends for those caring for the adult patient who has a tracheostomy tube. The first section of the article is an overview of the indications, surgical techniques, and types of tubes used in the critical care setting. The second section examines specific aspects of nursing care of patients who have tracheostomy tubes. PMID- 15358390 TI - The pragmatics of feeding the pediatric patient with acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) represents the ultimate pulmonary response to a wide range of injuries, from septicemia to trauma. Optimal nutrition is vital to enhancing oxygen delivery, supporting adequate cardiac contractility and respiratory musculature, eliminating fluid and electrolyte imbalances, and supporting the proinflammatory response. Research is providing a better understanding of nutrients that specifically address the complex physiologic changes in ARDS. This article highlights the pathophysiology of ARDS as it relates to nutrition, relevant nutritional assessment, and important enteral and parenteral considerations for the pediatric patient who has ARDS. PMID- 15358391 TI - Monitoring carbon dioxide in critical care: the newest vital sign? AB - Carbon dioxide (CO(2)) monitoring can yield substantial information about cardiac and pulmonary function. Because capnography is not complicated, it is relatively easy to apply in a wide variety of clinical settings. A new area of CO(2) measurement is sublingual CO(2). Although outcomes data are as yet unavailable for this technology, its ease of use makes it an attractive monitoring tool to assess severity of illness and predict patients' responses to therapy. This article describes the pathophysiology and clinical applications of the technologies and describes why they may well be "the newest vital signs." PMID- 15358393 TI - Quality of life: patients and doctors don't always agree: a meta-analysis. AB - OBJECTIVE: In addition to traditional clinical markers, quality-of-life assessment can be helpful to estimate the well-being of patients. Discrepancies in perception of well-being between physicians and patients may interfere with the effectiveness of treatment. A systematic review and meta-analysis were performed to explore the (dis-)agreement in quality-of-life assessments between patients and physicians. STUDY DESIGN AND SETTING: Data on the proportion agreement of paired observations were collected from Medline, Embase, Psychlit, and Social Abstracts. RESULTS: Of the 1,316 articles found, six met the selection criteria, four studied the proportion agreement between children and physicians, and all six the proportion agreement between parents and physicians. None examined the magnitude of over- or underestimation by physicians. The agreement was lower in the more subjective domains (0.54-0.77) in comparison to the more objective domains (0.79-0.94). CONCLUSION: Quality-of-life assessment should be integrated in clinical practice. During long-term treatment the perception of the patients' well-being by physicians and patients themselves can easily diverge from each other, resulting in misunderstandings about the treatment and its usefulness in relation to perceived quality of life, and may even become the base for noncompliance. PMID- 15358394 TI - The case-combined-control design was efficient in detecting gene-environment interactions. AB - OBJECTIVE: The interest in studying gene-environment (GxE) interaction is increasing for complex diseases. A design combining both related and unrelated controls (e.g., population-based and siblings) is proposed to increase the power to detect GxE interaction. STUDY DESIGN AND SETTING: We used simulations to assess the efficiency of the case-combined-control design relative to a classical case-control study under a variety of assumptions. RESULTS: The case-combined control design appears more efficient and feasible than a classical case-control study for detecting interaction involving rare exposures and/or genetic factors. The number of available sibling controls per case and the frequencies of the risk factors are the most important parameters for determining relative efficiency. Relative efficiencies decrease as the frequency of the gene (G) increases. A positive correlation in exposure (E) between siblings decreases relative efficiency. CONCLUSIONS: Although the case-combined-control design may not be efficient for common genes with moderate effects, it appears to be a useful alternative in certain situations where classical approaches remain unrealistic. PMID- 15358395 TI - Time-dependent bias was common in survival analyses published in leading clinical journals. AB - OBJECTIVE: In survival analysis, "baseline immeasurable" time-dependent factors cannot be recorded at baseline, and change value after patient observation starts. Time-dependent bias can occur if such variables are not analyzed appropriately. This study sought to determine the prevalence of such time dependent bias in highly-cited medical journals. STUDY DESIGN AND SETTING: We searched Medline databases to identify all observational studies that used a survival analysis in American Journal of Medicine, Annals of Internal Medicine, Archives of Internal Medicine, British Medical Journal, Chest, Circulation, Journal of the American Medical Association, Lancet, and New England Journal of Medicine between 1998 and 2002. Studies with "baseline immeasurable" time dependent factors were susceptible to time-dependent bias if a time-dependent covariate analysis was not used. RESULTS: Of 682 eligible studies, 127 (18.6%, 95% CI 15.8-21.8%) contained a "baseline immeasurable" time-dependent factor and 52 (7.6% [5.8-9.9%] of all survival analyses/40.9% [32.3-50.0%] of studies with a time-dependent factor) were susceptible to time-dependent bias. In 35 studies (5.1% [3.7-7.1%]/27.6% [20.5-35.9%]), the bias affected a variable highlighted in the study abstract and correction of the bias could have qualitatively changed the study's conclusion in over half of studies. CONCLUSION: In medical journals, time-dependent bias is concerningly common and frequently affects key factors and the study's conclusion. PMID- 15358396 TI - Meta-regression detected associations between heterogeneous treatment effects and study-level, but not patient-level, factors. AB - OBJECTIVE: Two investigations evaluate Bayesian meta-regression for detecting treatment interactions. STUDY DESIGN AND SETTING: The first compares analyses of aggregate and individual patient data on 1,860 subjects from 11 trials testing angiotensin converting enzyme (ACE) inhibitors for nondiabetic kidney disease. The second explores meta-regression for detecting treatment interaction on 671 covariates, including the baseline risk, from 232 meta-analyses of binary outcomes compiled from the Cochrane Collaboration and the medical literature. RESULTS: In the ACE inhibitor study, treatment effects were homogeneous so meta regression identified no interactions. Analysis of individual patient data using a multilevel model, however, discovered that treatment reduced glomerular filtration rate (GFR) more among patients with higher baseline proteinuria. The second investigation found meta-regression most effective for detecting treatment interactions with study-level factors in meta-analyses with >10 studies, heterogeneous treatment effects, or significant overall treatment effects. Under all three conditions, 46% of meta-regressions produced strong interactions (posterior probability >0.995) compared with 6% otherwise. Baseline risk was associated with the odds ratio in 6% of meta-analyses, half the rate found using maximum likelihood. CONCLUSION: Meta-regression can detect interactions of treatment with study-level factors when treatment effects are heterogeneous. Individual patient data are needed for patient-level factors and homogeneous effects. PMID- 15358397 TI - Repeated-measures modeling improved comparison of diagnostic tests in meta analysis of dependent studies. AB - OBJECTIVE: Current methods for meta-analysis of diagnostic tests do not allow utilizing all the information from papers in which several tests have been studied on the same patient sample. We demonstrate how to combine several studies of diagnostic tests, where each study reports on more than one test and some tests (but not necessarily all of them) are shared with other papers selected for the meta-analysis. We adopt statistical methodology for repeated measurements for the purpose of meta-analysis of diagnostic tests. STUDY DESIGN AND SETTING: The method allows for missing values of some tests for some papers, takes into account different sample sizes of papers, adjusts for background and confounding factors including test-specific covariates and paper-specific covariates, and accounts for correlations of the repeated measurements within each paper. It does not need individual-level data, although it can be modified to use them, and uses the two-by-two table of test results vs. gold standard. RESULTS: The results are translated from diagnostic odds ratios (DOR) to more clinically useful measures such as predictive values, post-test probabilities, and likelihood ratios. Models to capture between-study variation are introduced. The fit and influence of specific studies on the regression can be evaluated. Furthermore, model-based tests for homogeneity of DORs across papers are presented. CONCLUSION: The use of this new method is illustrated using a recent meta-analysis of the D-dimer test for the diagnosis of deep venous thrombosis. PMID- 15358398 TI - Null bar and null zone are better than the error bar to compare group means in graphs. AB - THE PROBLEM: Conventional graphs often include error bars around the group means. Regardless of what these bars depict, they are uninformative as to whether a difference between the groups is statistically significant. THE SOLUTION: This article suggests plotting the null bar or null zone: that is, the range or area in which the means of the two groups fall if the null hypothesis of no difference cannot be rejected. The bar and zone are simply derived from the confidence interval around the difference, obtained from the t-test. PMID- 15358399 TI - A scoring system identified near-miss maternal morbidity during pregnancy. AB - OBJECTIVE: The objective of this study was to develop a scoring system for identifying women with near-miss maternal morbidity, and differentiating these women from those with severe but not life-threatening conditions. STUDY DESIGN AND SETTING: The study was conducted at the University of Illinois Medical Center at Chicago (UIMC), which is a tertiary care hospital with approximately 2,220 births per year. UIMC is in a major urban area serving a predominantly African American and Latina population. This article focuses on five clinical factors: organ failure (>/=1 system), extended intubation (>12 hr), ICU admission, surgical intervention, and transfusion (>3 units), grouped into several scoring system alternatives. The total score on each scoring system was calculated as the weighted sum of the clinical factors present for each woman. RESULTS: The five factor scoring system had the highest specificity (93.9%), but the four-factor scoring system, which eliminated organ system failure for simplification of data collection, still had a specificity of 78.1%. CONCLUSION: Near-miss morbidities identified using the scoring systems presented can be incorporated into clinical case review and epidemiologic studies to enhance the monitoring of obstetric care and to improve estimates of the incidence of life-threatening complications in pregnancy. PMID- 15358400 TI - Comorbidity independently predicted death in older prostate cancer patients, more of whom died with than from their disease. AB - OBJECTIVE: The purpose of this study was to examine the proportion of men who died from prostate cancer (PrCa) vs. with PrCa and assess the comorbid conditions associated with other-cause deaths. STUDY DESIGN AND SETTING: We identified all male decedents aged >/=65 years in Jefferson County, AL, in 1993-1995. By crosslinking three databases (death certificate, Medicare, and Veteran's Administration), we identified men whose deaths might have been caused by PrCa. We abstracted and reviewed medical records to rate comorbid conditions and determine whether or not death was due to PrCa. RESULTS: Of 561 men with a premortem diagnosis of PrCa, 42% died from PrCa and 53% died with PrCA; 50.2% of blacks died from PrCa vs. 36.9% of Whites. Other factors related to dying with PrCa included older age at death and a serious, or very serious, comorbid condition. Treatment did not have an independent effect on cause of death (i.e., death with vs. from PrCa). CONCLUSIONS: Comorbidity was an independent predictor of dying with PrCa, even after adjustment for ethnicity, age, and treatment. Given the as yet unproven benefit of PrCa screening, our results extend the body of information relevant to the screening decision; among men dying with a diagnosis of PrCa, only about 1/3 to 1/2 died from the disease. PMID- 15358401 TI - Complaint-severity and cervical spine problems successfully classified patients with shoulder complaints. AB - OBJECTIVE: To construct a classification of patients with shoulder complaints based on their physical examination. To investigate (1) the interobserver reliability, (2) to what extent the setting in which the patients were recruited, and demographic and clinical characteristics are related to the classification. STUDY DESIGN AND SETTING: Data from 132 patients with shoulder complaints recruited in various health care settings in The Netherlands were examined. Two observers independently performed a physical examination of the cervical spine and shoulder joint. A nonmetric multidimensional scaling procedure was performed for each observer separately. The interobserver reliability of both observers was computed. Differences between setting, demographic and clinical characteristics, and the resulting dimensions were investigated. RESULTS: For both observers two dimensions (severity of complaints of the shoulder joint, and severity of problems of the cervical spine) were sufficient to classify all patients. Agreement between the two observers was good (r=0.84) to moderate (r=0.69). Patients with neck pain in history taking showed higher scores on both dimensions. CONCLUSION: Despite moderate interobserver agreement for each variable from physical examination found in previous studies, observers agree on the scores of the patients on the relevant dimensions. Given the limited number of effective treatments available to the general practitioner, a more sophisticated classification system seems unnecessary. PMID- 15358402 TI - Identifying parents in pharmacy data: a tool for the continuous monitoring of drug exposure to unborn children. AB - OBJECTIVE: This study aimed to develop and validate a method for retrospectively identifying parents in pharmacy data during pregnancy. STUDY DESIGN AND SETTING: The principle of the method was to select all children 0-2 years in pharmacy records, and to consider men/women 15-50 years older with the same address as fathers/mothers. RESULTS: Applying this method to the records of all 4 pharmacies in 1 town (33,000 inhabitants) resulted in identification of 807 fathers and 765 mothers, corresponding with 68.5% of all fathers, and 64.9% of all mothers from the town. Additionally, the method was applied to one selected pharmacy, resulting in 151 fathers and 170 mothers. Validation criterions, evaluated by pharmacy employees and GPs, disproved one of these fathers (0.7% of all identified fathers) and one mother (0.6%). CONCLUSION: We conclude that automatic retrospective identification of parents in pharmacy data is feasible in a valid way. The main limitation is that not all parents were found, possibly resulting in selection bias. PMID- 15358403 TI - A community survey demonstrated cohort differences in the lifetime prevalence of self-reported head injury. AB - OBJECTIVE: The objective of the study was to assess the lifetime prevalence and validity of self-reported head injury. STUDY DESIGN AND SETTING: Analysis of a cross-sectional community survey, stratified by birth cohort. Seven thousand four hundred eighty-eight residents of Canberra and Queanbeyan, 3,678 males and 3,810 females, representing three cohorts aged in their 20s, 40s and 60s at the time of the survey, were randomly selected from the Australian Electoral Roll. RESULTS: The lifetime prevalence of head injury with at least 15 min loss of consciousness ranged from 5.6 to 6.0% across the three cohorts. Self-reported head injury was associated with symptoms of traumatic brain injury or postconcussion syndrome. Reported head injury did not differ across the birth cohorts, contrary to expectations that prevalence rates would increase with age. Analysis of age at first head injury showed significant differences between cohorts in the reported prevalence at age 20, with estimates considerably higher for the youngest cohort. CONCLUSION: Although the data may reflect real increases in the risk of head injury, it may be that self-reported measures of lifetime prevalence of head injury underestimate prevalence in older cohorts. PMID- 15358404 TI - Lower ambient temperature was associated with an increased risk of hospitalization for stroke and acute myocardial infarction in young women. AB - OBJECTIVE: Results of investigations into the impact of seasonal variation on the incidence of cardiovascular diseases (CVD) have been inconsistent. Using the WHO Collaborative Study of CVD and Steroid Hormone Contraception database, we attempted to examine the relationship between variation in three climatic variables and risk of hospitalization for venous thromboembolism (VTE), arterial stroke, and acute myocardial infarction (AMI). STUDY DESIGN AND SETTING: We compared the monthly mean temperature, rainfall, and humidity with rates of hospitalized VTE, stroke, and AMI among young women aged 15-49 from 17 different countries in Africa, Asia, Europe, Latin America, and the Caribbean by using a negative binomial regression model. RESULTS: The study included 1146, 2,269, and 369 cases of VTE, stroke, and AMI, respectively. Significant associations between temperature and hospital admission rates of stroke and AMI, but not VTE, were apparent. Lagging the effects of temperature suggested that these effects were relatively acute, within a period of a month. CONCLUSION: These data may help in understanding the mechanisms whereby stroke and AMI events are triggered. PMID- 15358405 TI - Prognostic index for stroke mortality. PMID- 15358406 TI - Short vs. long bed rest after uncomplicated acute myocardial infarction: a systematic review and meta-analysis. PMID- 15358408 TI - International differences in major depression prevalence: what do they mean? PMID- 15358410 TI - United Kingdom National Health Service, Cancer Services Collaborative "Improvement Partnership", Redesign of Cancer Services: A National Approach. PMID- 15358411 TI - Protein secretion in Legionella pneumophila and its relation to virulence. AB - Protein secretion is a universal process of fundamental importance for various aspects of cell physiology including the infection of a host organism by a bacterial pathogen. Many Gram-negative pathogens export virulence-associated proteins across one or two cell membranes to their place of action using a wide plethora of secretory pathways with the objective of infecting the host. For Legionella pneumophila, a facultative intracellular, human pathogen which is ubiquitously found in natural and artificial aquatic environments, two major secretory pathways known to be involved in virulence have been described. These are the PilD-dependent Lsp type II secretion pathway and the type IV secretion system encoded by the dot/icm genes. In addition, a second type IV system, with high sequence similarity to the Agrobacterium tumefaciens VirB system for conjugal transfer of oncogenic DNA, is present. Albeit dispensable for intracellular growth, this type IV system is important for efficient host cell infection at lower temperatures. Further more, evidence exists for the presence of at least one type I secretion system in L. pneumophila as well as for the presence of a twin arginine dependent translocation (Tat) pathway. This is a recently detected, signal peptide-dependent, secretion pathway complementary to the well-known Sec-dependent pathway for protein transport across the cytoplasmic membrane. PMID- 15358412 TI - Cholesterol and the activity of bacterial toxins. AB - Cholesterol may affect the activity of microbial toxins in a direct, specific way, or it may exert indirect effects because of its role in membrane fluidity, membrane line tension, and in the stabilization of rafts in the cytoplasmic membrane. The thiol-activated toxins of gram-positive bacteria, and the cytolysin of Vibrio cholerae are presented as examples of specific toxin-cholesterol interaction. Several mechanisms of indirect effects of cholesterol are discussed using examples such as Staphylococcus aureus alpha-hemolysin, aerolysin, and diphtheria toxin. PMID- 15358413 TI - Low pH-induced membrane fatty acid alterations in oral bacteria. AB - Four oral bacterial strains, of which two are considered aciduric and two are considered acid-sensitive, were grown under glucose-limiting conditions in chemostats to determine whether their membrane fatty acid profiles were altered in response to environmental acidification. Streptococcus gordonii DL1, as well as the aciduric strains S. salivarius 57.I, and Lactobacillus casei 4646 increased the levels of mono-unsaturated membrane fatty acids. The non-aciduric strain S. sanguis 10904 did not alter its membrane composition in response to pH values examined here. Thus, in response to low pH, aciduric oral bacteria alter their membrane composition to contain increased levels of long-chained, mono unsaturated fatty acids. This suggests that membrane fatty acid adaptation is a common mechanism utilized by bacteria to withstand environmental stress. PMID- 15358414 TI - Isolation and characterization of genes encoding polycyclic aromatic hydrocarbon dioxygenase from acenaphthene and acenaphthylene degrading Sphingomonas sp. strain A4. AB - Sphingomonas sp. strain A4 is capable of utilizing acenaphthene and acenaphthylene as sole carbon and energy sources, but it is unable to grow on other polycyclic aromatic hydrocarbons (PAHs). The genes encoding terminal oxygenase components of ring-hydroxylating dioxygenase (arhA1 and arhA2) were isolated from this strain by means of the ability to oxidize indole to indigo of the Escherichia coli clone containing electron transport proteins from phenanthrene-degrading Sphingobium sp. strain P2. The translated products of arhA1 and arhA2 exhibited moderate sequence identity (less than 56%) to large and small subunits of dioxygenase of other ring-hydroxylating dioxygenases. Biotransformation with recombinant E. coli clone revealed the broad substrate specificity of this oxygenase toward several PAHs including acenaphthene, acenaphthylene, naphthalene, phenanthrene, anthracene and fluoranthene. Southern hybridization analysis revealed the presence of a putative arhA1 homologue on a locus different from that of the arhA1 gene. Insertion inactivation of the arhA1 gene in strain A4 suggested that the gene but not the putative homologue one was involved in the degradation of acenaphthene and acenaphthylene in this strain. PMID- 15358415 TI - A re-appraisal of the diversity of the methanogens associated with the rumen ciliates. AB - The diversity of methanogenic archaea associated with different species of ciliated protozoa in the rumen was analysed. Partial fragments of archaeal SSU rRNA genes were amplified from DNA isolated from single cells from the rumen protozoal species Metadinium medium, Entodinium furca, Ophryoscolex caudatus and Diplodinium dentatum. Sequence analysis of these fragments indicated that although all of the new isolates clustered with sequences previously described for methanogens, there was a difference in the relative distribution of sequences detected here as compared to that of previous work. In addition, many of the novel sequences, although clearly of archaeal origin have relatively low identity to the sequences in database which are most closely related to them. PMID- 15358416 TI - The targeted inactivation of polyketide synthase mycAV in the mycinamicin producer, Micromonospora griseorubida, and a complementation study. AB - Mycinamicin is a 16-membered macrolide antibiotic produced by Micromonospora griseorubida A11725, which shows strong antimicrobial activity against gram positive bacteria. Recently, the nucleotide sequences of the mycinamicn biosynthetic gene cluster in M. griseorubida have been completely determined. Mycinamicin non-producer M7A21 was isolated by mycAV inactivation, which encodes the module 7 of mycinamicin polyketide synthase (PKS) required for the biosynthesis of the mycinamicin biosynthetic intermediate protomycinolide-IV (PML IV). When the bioconversion to mycinamicin II (M-II) from PML-IV was performed using M7A21 and the feeding culture method, the productivity of M-II was the same as that of M-II in wild-type strain A11725. p446M7 containing mycAV was constructed using the Escherichia coli-Streptomyces shuttle vector pGM446. The mycinamicin productivity of M7A21 was restored by the introduction of p446M7 into the M7A21 cell, but almost all p446M7 was integrated into the chromosome of M7A21 because the plasmid was unstable in M7A21. The feeding culture and the introduction of the complement gene for M7A21 would be powerful tools to perform combinatorial biosynthesis for the production of new macrolide antibiotics. PMID- 15358417 TI - Molecular characterization of extraintestinal pathogenic Escherichia coli (ExPEC) pathogenicity islands in F165-positive E. coli strain from a diseased animal. AB - Septicemic Escherichia coli 4787 (O115: K-: H51: F165) of porcine origin possess gene clusters related to extraintestinal E. coli fimbrial adhesins. This strain produces two fimbriae: F165(1) and F165(2). F165(1) (Prs-like) belongs to the P fimbrial family, encoded by foo operon and F165(2) is a F1C-like encoded by fot operon. Data from this study suggest that these two operons are part of two PAIs. PAI I(4787) includes a region of 20 kb, which not only harbors the foo operon but also contains a potential P4 integrase gene and is located within the pheU tRNA gene, at 94 min of the E. coli chromosome. PAI II(4787) includes a region of over 35 kb, which harbors the fot operon, iroBCDEN gene clusters, as well as part of microcin M genes and nonfunctional mobility genes. PAI II(4787) is found between the proA and yagU at 6 min of the E. coli chromosome. PMID- 15358418 TI - Characterization of the second long polar (LP) fimbriae of Escherichia coli O157:H7 and distribution of LP fimbriae in other pathogenic E. coli strains. AB - A second region containing five genes homologous to the long polar fimbrial operon of Salmonella enterica serovar Typhimurium is located in the chromosome of enterohemorrhagic Escherichia coli (EHEC) O157:H7. A non-fimbriated E. coli K-12 strain carrying the cloned EHEC lpf (lpf2) genes expressed thin fibrillae-like structures on its surface and displayed reduced adherence to tissue culture cells. Neither mutation in the lpfA2 gene in either the parent or lpfA1 mutant strains showed an effect in adherence or in the formation of A/E lesions on HeLa cells. lpfA2 isogenic mutant strains adhere to Caco-2 cells almost as well as the wild-type at 5 h, but they were deficient in adherence at early time points. A collection of diarrheagenic E. coli strains were investigated for the presence of lpfA1 and lpfA2 and results showed that these genes are present in specific serogroups which are phylogenetically related. Our results suggest that LP fimbriae 2 may contribute to the early stages of EHEC adhesion and that genes encoding the major LP fimbrial subunits are present in a small group of EHEC and EPEC serotypes. PMID- 15358419 TI - Novel strains of Salmonella typhimurium as potential vectors for gene delivery. AB - DNA vaccines are known to induce long-term antigen specific cellular responses. We tested two new strains of Salmonella typhimurium, one carrying a mutation in a SPI-2 gene and the aroC-gene and another carrying mutations in the sifA- and aroC genes, as potential DNA vaccine delivery vehicles. We compared them with the SL7207 strain and found that the new strains were more invasive, and that they were efficient mediators of gene transfer in vitro using EGFP as reporter gene. We tested the ability of the new strains to survive within the spleen, liver and mesenteric lymph nodes and evaluated their safety in C57/BL/6J mice. PMID- 15358420 TI - Characterization of extended-spectrum-beta-lactamase (CTX-M-15)-producing strains of Salmonella enterica isolated in France and Senegal. AB - Three clinical isolates of Salmonella enterica recovered between 2000 and 2003 in France and Senegal were found to produce extended-spectrum beta-lactamase CTX-M 15. The two isolates from Senegal were recovered from stool of a hospitalized patient with gastroenteritis in 2000 and from an urine specimen of an out-patient with urinary tract infection in 2001. These S. enterica isolates belonged to serotype Kentucky and were clonally related as determined by pulsed-field gel electrophoresis and automated ribotyping. The third isolate of serotype Typhimurium was recovered from a patient hospitalized in France for an acute gastroenteritis acquired in Lebanon. The bla(CTX-M-15) gene was located on two different transferable plasmids, one of which also carried bla(TEM-1), bla(OXA 30), aminoglycoside-, tetracycline-, and sulfamethoxazole-trimethoprim resistance genes. ISEcp1 element was found to be located upstream of bla(CTX-M-15) in the same position as reported previously in CTX-M-15-producing Escherichia coli from India and Turkey. This is the first report of bla(CTX-M-15) in the genus Salmonella. PMID- 15358421 TI - Characterization of the in vitro antimycotic activity of a novel killer protein from Williopsis saturnus DBVPG 4561 against emerging pathogenic yeasts. AB - A novel killer toxin, labelled as KT4561, secreted by Williopsis saturnus DBVPG 4561, was found to possess a wide antimycotic activity against strains of Candida glabrata, Issatchenkia orientalis and Pichia guillermondii. KT4561 was precipitated by ethanol and purified by ion-exchange chromatography. The active protein migrated as a single band in SDS-PAGE and was characterized by a molecular weight of approximately 62 kDa. Purified KT4561 was active across wide ranges of temperature (5-45 degrees C) and pH (4.5-8.0) and displayed a rapid decrease in viability of yeast cells after 4-8 h. The in vitro activity of KT4561 against 102 yeast isolates (79% of clinical origin) was determined: MIC(50) and MIC(90) of strains were 0.08 and 0.15 microg/ml for C. glabrata, 0.03 and 0.23 microg/ml for I. orientalis and 1.50 and 2.25 microg/ml for P. guilliermondii. Comparative susceptibility tests showed that a high number of strains used in the present study were insensitive to selected azole and polyene antibiotics. The present study demonstrated the potential of KT4561 to be applied as novel control agent against pathogenic yeasts. PMID- 15358422 TI - Biochemical and molecular characterization of alpha-ketoisovalerate decarboxylase, an enzyme involved in the formation of aldehydes from amino acids by Lactococcus lactis. AB - In this paper, we report for the first time on the identification, purification, and characterization of the alpha-ketoisovalerate decarboxylase from Lactococcus lactis, a novel enzyme responsible for the decarboxylation into aldehydes of alpha-keto acids derived from amino acid transamination. The kivd gene consisted of a 1647 bp open reading frame encoding a putative peptide of 61 kDa. Analysis of the deduced amino acid sequence indicated that the enzyme is a non-oxidative thiamin diphosphate (ThDP)-dependent alpha-keto acid decarboxylase included in the pyruvate decarboxylase group of enzymes. The active enzyme is a homo-tetramer that showed optimum activity at 45 degrees C and at pH 6.5 and exhibited an inhibition pattern typical for metal-dependant enzymes. In addition to Mg(2+), activity was observed in presence of other divalent cations such as Ca(2+), Co(2+) and Mn(2+). The enzyme showed the highest specific activity (80.7 Umg(-1)) for alpha-ketoisovalerate, an intermediate metabolite in valine and leucine biosynthesis. On the other side, decarboxylation of indole-3-pyruvate and pyruvate only could be detected by a 100-fold increase in the enzyme concentration present in the reaction. PMID- 15358423 TI - Biodiversity amongst cultivable polycyclic aromatic hydrocarbon-transforming bacteria isolated from an abandoned industrial site. AB - The characterisation of a microbial community of a polycyclic aromatic hydrocarbons (PAHs) contaminated site (formerly Carbochimica, Trento, Italy) was carried out. A preliminary evaluation of the heterogeneity and the metabolic activity of the microbial community were attempted by denaturing gradient gel electrophoresis (DGGE) and reverse transcriptase-denaturing gel electrophoresis (RT-DGGE). The presence of a heterogeneous and metabolically active microbial community was found. To evaluate the PAH-transforming potential of the soil bacterial community, enrichment cultures were set up. Taxonomically diverse bacteria, showing different biochemical PAH-transforming pathways were obtained. Some of the isolates showed not nah-homologous PAH-transforming genotypes. PMID- 15358424 TI - Oxidation of 3,4-dehydro-D-proline and other D-amino acid analogues by D-alanine dehydrogenase from Escherichia coli. AB - 3,4-Dehydro-DL-proline is a toxic analogue of L-proline which has been useful in studying the uptake and metabolism of this key amino acid. When membrane fractions from Escherichia coli strain UMM5 (putA1::Tn5 proC24) lacking both L proline dehydrogenase and L-Delta(1)-pyrroline-5-carboxylate reductase were incubated with 3,4-dehydro-DL-proline, pyrrole-2-carboxylate was formed. There was no enzyme activity with 3,4-dehydro-L-proline, but activity was restored after racemization of the substrate. Oxidation of 3,4-dehydro-DL-proline by membrane fractions from strain UMM5 was induced by growth in minimal medium containing D- or L-alanine, had a pH optimum of 9, and was competitively inhibited by D-alanine. An E. coli strain with no D-alanine dehydrogenase activity due to the dadA237 mutation was unable to oxidize either 3,4-dehydro-D proline or D-alanine, as were spontaneous Dad(-) mutants of E. coli strain UMM5. Membrane fractions containing D-alanine dehydrogenase also catalyzed the oxidation of D-2-aminobutyrate, D-norvaline, D-norleucine, cis-4-hydroxy-D proline, and DL-ethionine. These results indicate that d-alanine dehydrogenase is responsible for the residual 3,4-dehydro-DL-proline oxidation activity in putA proC mutants of E. coli and provide further evidence that this enzyme plays a general role in the metabolism of D-amino acids and their analogues. PMID- 15358425 TI - New olivosyl derivatives of methymycin/pikromycin from an engineered strain of Streptomyces venezuelae. AB - A mutant strain of Streptomyces venezuelae was engineered by deletion of the entire gene cluster related to biosynthesis of the endogenous deoxysugar (TDP-D desosamine) and replacement with genes required for biosynthesis of an intermediate sugar (TDP-4-keto-6-deoxy-D-glucose) or an exogenous sugar (TDP-D olivose), from the oleandomycin and urdamycin deoxysugar pathways. The 'sugar flexible' glycosyltransferase (DesVII) was able to attach the intermediate sugar and the new sugar to both 12- and 14-membered macrolactones thus producing quinovose or olivose glycosylated 10-deoxymethynolide and narbonolide, respectively. In addition, hydroxylated analogs of the new metabolites were detected. These results demonstrate a successful attempt of engineering the deoxysugar pathway for generation of novel hybrid macrolide antibiotics. PMID- 15358426 TI - Development of an in vivo Himar1 transposon mutagenesis system for use in Streptococcus equi subsp. equi. AB - Streptococcus equi subsp. equi is the causative agent of the equine disease strangles. In this study we describe the development of an in vivo Himar1 transposon system for the random mutagenesis of S. equi and, potentially, other Gram-positive bacteria. We demonstrate efficient and random transposition of a modified mini-transposon onto the chromosome by Southern blot analysis and insertion site sequencing. Non-haemolytic mutants were isolated at a frequency of 0.2%, and acapsular mutants at a frequency of 0.04%. Taken together, these data demonstrate that in vivo Himar1 mutagenesis can be used for genomic-scale mutational analysis of S. equi, and is likely to be applicable to the study of other streptococci. PMID- 15358427 TI - Gram-positive merA gene in gram-negative oral and urine bacteria. AB - Clinical mercury resistant (Hg(r)) Gram-negative bacteria carrying Gram-positive mercury reductase (merA)-like genes were characterized using DNA-DNA hybridization, PCR and sequencing. A PCR assay was developed which discriminated between the merA genes related to Staphylococcus and those related to the Bacillus/Streptococcus merA genes by the difference in size of the PCR product. DNA sequence analysis correlated with the PCR assay. The merA genes from Acinetobacter junii, Enterobacter cloacae and Escherichia coli were sequenced and shared 98-99% identical nucleotide (nt) and 99.6-100% amino acid identity with the Staphylococcus aureus MerA protein. A fourth merA gene, from Pantoeae agglomerans, was partially sequenced (60%) and had 99% identical nt and 100% amino acid identity with the Streptococcus oralis MerA protein. All the Hg(r) Gram-negative bacteria transferred their Gram-positive merA genes to a Gram positive Enterococcus faecalis recipient with the resulting transconjugants expressing mercury resistance. These Gram-positive merA genes join Gram-positive tetracycline resistance and Gram-positive macrolide resistance genes in their association with mobile elements which are able to transfer and express in Gram negative bacteria. PMID- 15358428 TI - Cell shape regulation and co-translocation of actin and adenosyl homocysteinase in response to intermediate hypertonicity. AB - Hypertonic stimulation induced association of S-adenosyl-L-homocysteine hydrolase (SAHH) with the F-actin-rich cell cortex in Dictyostelium. At intermediate, but not higher, levels of hypertonicity, SAHH further translocated from the cortex to the cytosol in company with a fraction of actin and cofilin. At the same time the cells rounded up. Acidification of the cells stimulated both the cell rounding and the translocation of actin and SAHH, whereas alkalinization retarded these responses, suggesting that cellular pH is involved in their control. On the other hand, mutant analysis suggested that neither cGMP signaling nor conventional myosin is required. PMID- 15358429 TI - Molecular typing of Yersinia frederiksenii strains by means of 16s rDNA and gyrB genes sequence analyses. AB - The phenospecies Yersinia frederiksenii is known to comprise three genospecies, indistinguishable on the basis of phenotypic characteristics. In previous works, 13 strains, identified biochemically as Y. frederiksenii, were characterized using Multilocus enzyme electrophoresis and Ribotyping. In order to elucidate the phylogenetic position of these strains we performed their molecular typing by means of 16S rDNA and gyrB sequences analyses. Results demonstrated that gyrB is a better phylogenetic marker than 16S rDNA. The classification achieved by gyrB sequences analyses was in agreement with results obtained with more laborious methods. Moreover, a good phylogenetic identification could be reached also with partial gyrB sequences of only 350 bp. PMID- 15358430 TI - Mycobacterium leprae infection of human Schwann cells depends on selective host kinases and pathogen-modulated endocytic pathways. AB - Mycobacterium leprae, an obligate intracellular pathogen, shows a unique tropism for Schwann cells (SC). This leads to the peripheral neuropathy disorder observed in leprosy. In this study, we investigated signal transduction events and the intracellular fate of M. leprae during the interaction of the microorganism with SC. First, we demonstrated that the human schwannoma cell line ST88-14 readily phagocytized the bacteria as observed by time-lapse microscopy, actin staining and electron microscopy. The effect of specific kinase inhibitors on M. leprae internalization was then investigated showing that functional protein tyrosine kinase, calcium-dependent protein kinase and phosphatidylinositol 3-kinase, but not cAMP-dependent protein kinase are essential for phagocytosis of the bacteria. Similar results were obtained when irradiated and live bacteria were compared and when M. leprae was pre-coated with recombinant histone-like-protein/laminin binding protein, a bacterial adhesin. In addition, experiments were performed to analyze the bacterial trafficking within the endosomal network by labeling the acidified intracellular compartments of M. leprae-infected SC with the Lysotracker acidotrophic probe. Acidification of vesicles containing live M. leprae was minimal in both RAW murine macrophages and SC, although phagosomes containing heat-killed bacteria seem to follow normal endocytic maturation. These data indicate that the invading bacteria interfere with normal endocytic pathway maturation of bacteria-containing phagosomes within SC. PMID- 15358431 TI - Widespread activation of antibiotic biosynthesis by S-adenosylmethionine in streptomycetes. AB - The effect of S-adenosylmethionine (SAM) on the production of various antibiotics was investigated to determine whether SAM-dependent methylation is required in biosynthetic pathways of antibiotics. Pristinamycin II(B) and granaticin do not require SAM-dependent methylation in their biosynthesis pathways, and production of these two antibiotics was increased about 2-fold when a low concentration (50 and 10 microM, respectively) of SAM was treated; in contrast, oleandomycin and avermectin B1a require SAM as a methyl donor in their biosynthesis, and production of these two antibiotics was increased 5-fold and 6-fold, depending on the SAM concentration within a certain range. We also found that the transcription of a pathway-specific regulator, gra-ORF9, was activated by exogenous SAM treatment. Production of oleandomycin and avermectin B1a was decreased by using a methyltransferase inhibitor, sinefungin, but the production levels of these antibiotics were restored to the control level by simultaneously adding SAM and sinefungin. Interestingly, we have found a similar stimulatory effect of S-adenosylhomocysteine (SAH), the methylation product of SAM, on antibiotic production in the four strains. Our results clearly demonstrate the widespread activation of antibiotic production using SAM in streptomycetes. PMID- 15358432 TI - GABAA receptor subunits identified in Paramecium by immunofluorescence confocal microscopy. AB - The presence of opioid, beta-adrenergic and cholinergic receptors has been demonstrated in ciliated protozoa, but little is known about gamma-aminobutyric acid (GABA) receptors. In this study we have analyzed the distribution of GABA(A) type receptor subunits in Paramecium. Confocal laser microscopy using antibodies specific for alpha(1)-, alpha(2)-, alpha(3)-, alpha(6)-, beta(2/3)-, gamma(2)-, epsilon-, lambda-, and theta-subunits showed that most receptors are aggregated in clusters and are distributed both on cell surface and in the cytoplasm. The intensity of labelling of the alpha(6)-, beta(2/3)- and gamma(2)-subunits was more intense than the alpha(1)-, epsilon-, and theta-subunits, suggesting that the former are present in higher concentrations than the latter. PMID- 15358433 TI - Polymorphisms and intronic structures in the 18S subunit ribosomal RNA gene of the fungi Scytalidium dimidiatum and Scytalidium hyalinum. Evidence of an IC1 intron with an His-Cys endonuclease gene. AB - The fungi Scytalidium dimidiatum (Nattrassia mangiferae synanamorph) and Scytalidium hyalinum are mainly encountered in (sub)tropical areas as plant pathogens and agents of human dermatomycosis. Because the classification and differentiation of these two species is unclear, we studied 22 S. dimidiatum and 15 S. hyalinum isolates in order to identify potential species-specific insertions and polymorphisms in the 18S subunit ribosomal gene. The presence of an IE intron in S. dimidiatum, together with a single polymorphism (A in S. dimidiatum, G in S. hyalinum) in the coding region, allowed us to differentiate these two species in most cases. Moreover, in one S. dimidiatum isolate we found a group IC1 intron containing a putative truncated His-Cys endonuclease gene. This enzyme shows strong similarity to the intronic homing endonuclease of Physarum polycephalum. Based on these results and our previous findings, we propose an evolutionary pathway for 18S rDNA S. dimidiatum insertions, implying independent events. PMID- 15358434 TI - Cultivation of Walsby's square haloarchaeon. AB - The square haloarchaea of Walsby (SHOW group) dominate hypersaline microbial communities but have not been cultured since their discovery 25 years ago. We show that natural water dilution cultures can be used to isolate members of this group and, once in pure culture, they can be grown in standard halobacterial media. Cells display a square morphology and contain gas vesicles and poly-beta hydroxybutyrate (PHB) granules. The 16S rRNA gene sequence was >99% identical to other SHOW group sequences. They prefer high salinities (23-30%), and can grow with a doubling time of 1-2 days in rich media. The ability to culture SHOW group organisms makes it possible to study, in a comprehensive way, the microbial ecology of salt lakes. PMID- 15358435 TI - Abstracts of the American Society for Therapeutic Radiology and Oncology 46th Annual Meeting, October 3-7, 2004, Atlanta, Georgia, USA. PMID- 15358437 TI - Role of stress in the pathogenesis of the metabolic syndrome. AB - Excess body fat, obesity, is one of the most common disorders in clinical practice. In addition, there is a clustering of several risk factors with obesity, including hypertension, glucose intolerance, diabetes mellitus, and hyperlipidemia, which is observed more frequently than by chance alone. This has led to the suggestion that these represent a single syndrome and is referred to as the Metabolic Syndrome. A growing body of evidence suggests that glucocorticoid secretion is associated with this complex phenotype. Continuously changing and sometimes threatening external environment may, when the challenge exceeds a threshold, activate central pathways that stimulate the adrenals to release glucocorticoids. In this review, we will discuss how such processes mediate a pathogenetic role in the Metabolic Syndrome. PMID- 15358438 TI - Distress proneness and cognitive decline in a population of older persons. AB - The association between distress proneness and cognitive decline was examined in older residents of a biracial community in Chicago. At baseline, participants completed four cognitive tests that yielded a global measure (baseline mean=101.2; standard deviation (SD)=7.8), and a brief measure of the tendency to experience negative emotions (mean=16.5; SD=6.7) based on the Neuroticism scale of the NEO Five-Factor Inventory. Cognitive testing was repeated twice at three year intervals. In mixed models that controlled age, sex, race, and education, for each point on the distress proneness scale, global cognitive score was 0.12 unit lower at baseline (p<0.001) and annual rate of decline increased by 0.01 unit (p=0.002), or about 2%. Thus, cognitive decline was about 30% faster in a person highly prone to distress (score=24, 90th percentile) compared to the one low in distress proneness (score=9, 10th percentile). This effect was unchanged after controlling for level of cognitive activity or excluding people with cognitive impairment at baseline, but it was reduced to a trend (p=0.059) after controlling for depressive symptoms. The results suggest that the tendency to experience psychological distress is associated with increased cognitive decline in old age. PMID- 15358439 TI - DHEAS and POMS measures identify cocaine dependence treatment outcome. AB - Early attrition is a significant problem in the treatment of cocaine dependence, but it is unclear why some patients succeed in treatment while others relapse or drop out of treatment without a demonstrated relapse. The goal of this study was to determine whether baseline levels of select hormones, including the adrenal hormone and excitatory neurosteroid dehydroepiandrosterone sulfate (DHEAS), would distinguish between treatment outcome groups. Based on the literature, completion of 90 days of treatment was established as a key outcome variable. METHODS: Quantitative urine levels of the cocaine metabolite benzoylecgonine (BE) and other substance of abuse analytes, plasma levels of DHEAS, DHEA, cortisol, and prolactin, and the profile of mood states (POMS) were serially measured in 38 male cocaine-dependent (DSM-IV) patients and in 28 controls of similar gender and age over a six month study. Exclusion criteria for the patients and controls included Axis I mood, anxiety or psychotic disorders. The patients could not manifest substance dependence except to cocaine. The patients and controls received remuneration for urine and blood collection. Blood samples for hormone levels were obtained between 8 and 10 a.m. on days 1, 14 and 21 of a 21-day inpatient treatment program and throughout 6 months of outpatient study visits at 45-day intervals. RESULTS: Attrition from treatment and study appointments occurred predominately at the junction between inpatient and outpatient programs. Forty percent of patients made the transition to outpatient treatment and remained abstinent and in treatment for a median of 103 days (ABST). Forty-two percent of patients dropped out of treatment during the inpatient stay or never returned after completing the inpatient program (DO) and 18% had a documented relapse either during, or within the first week after, the inpatient stay (REL). POMS total scores were elevated at treatment entry for both the ABST and DO groups. Plasma DHEAS levels in the DO patients were decreased compared to controls and increased in the ABST patients. POMS total scores for the REL patients at baseline were at control levels. Baseline cortisol levels were not statistically different between the outcome groups, though they were elevated for all cocaine patient groups. When treatment outcome was collapsed into whether patients completed (ABST) or did not complete 90 days of treatment (90N), ABST plasma DHEAS and cortisol were significantly elevated compared to the 90N patients and controls across the first 3 weeks of cocaine withdrawal. CONCLUSIONS: At treatment entry, each of the three patient outcome groups was identified by levels of circulating DHEAS and distressed mood. In the ABST patients, distressed mood during withdrawal may have been mitigated through antidepressant-like actions of enhanced endogenous DHEAS activity, thus contributing to improved abstinence and treatment retention. Patients, such as the DO group, with high levels of distressed mood at treatment entry and low DHEAS levels may benefit from adjunctive pharmacotherapy that targets DHEAS and POMS measures. Patients, such as the REL group, who lack distressed mood at treatment entry, may require intense application of motivational approaches plus residential treatment. PMID- 15358440 TI - Night sleep EEG and daytime sleep propensity in adult hypopituitary patients with growth hormone deficiency before and after six months of growth hormone replacement. AB - BACKGROUND: Hypopituitary patients with growth hormone deficiency (GHD) complain of reduced vitality, general fatigue, lack of concentration, irritability and reduced alertness during daytime. It is unclear whether these symptoms are primarily due to GH-deficiency and/or secondary to GHD related sleep impairments. Bi-directional interactions between the somatotropic system and human sleep patterns are well established. However, data on the effect of GH either in subjects without GHD or in patients with GHD under GH replacement therapy on the sleep electroencephalogram (EEG) are controversial. No reports exist about objective measures of daytime sleepiness in GH deficient patients before and during GH-therapy. OBJECTIVE: To assess the effects of GH on nocturnal and daytime sleep in adult patients with GHD before and during recombinant human GH (rhGH, Somatropin) replacement therapy. METHODS: Eighteen adult patients with GHD (4 women and 14 men) participated in the study. Mean age at the beginning of the study was 48.5 years (range 27-64 years). Ten patients were recruited from a double-blind, randomized placebo controlled trial over 6 months, followed by an open treatment period of 6 additional months (Group I). In all patients from this group, only the effects of the first 6 months of GH treatment were assessed. Eight additional patients were treated in an open study design for 6 months (Group II). Nocturnal sleep recordings and daytime sleep EEGs with a multiple sleep latency test were performed at baseline and after 6 months of additional GH replacement therapy. RESULTS: One patient dropped out due to side effects and was not included in sleep analysis. IGF-1 levels were increased in all patients, partially in a supraphysiologic range. Side effects were mainly mild but in one patient (from group II), general muscle pain led to interruption of the study. Therefore sleep analysis was only done in 17 patients. Sleep parameters were comparable to healthy control groups from the literature. GH substitution over 6 months did neither affect total sleep time nor times spent in different sleep stages. REM sleep density was also not changed. Daytime sleep propensity as measured by the multiple sleep latency test was not influenced by GH treatment. CONCLUSIONS: GH replacement does neither affect night sleep nor daytime sleep propensity in GH deficient hypopituitary adults. GH substitution has no sleep disturbing effect. PMID- 15358441 TI - Estradiol and the addition of progesterone increase the sensitivity to a neurosteroid in postmenopausal women. AB - The aim of this study was to compare the pharmacodynamic response to a neuroactive steroid, pregnanolone, before and during different hormonal settings of postmenopausal hormone replacement therapy (HRT), using natural progesterone. A second aim was to investigate whether the response to pregnanolone was associated with cyclicity in negative mood symptoms during treatment. Twenty six postmenopausal women with climacteric symptoms were administered HRT in a randomized, double blinded, placebo-controlled, crossover study. The women received 2 mg oral estradiol (E(2)) continuously during two 28-day cycles and 800 mg of vaginal progesterone or placebo sequentially for the last 14 days of each treatment cycle. Pharmacodynamic response to pregnanolone was assessed before treatment, and during the last week of each treatment cycle, by comparing the effects of intravenous pregnanolone (3alpha-hydroxy-5beta-pregnan-20-one) on saccadic eye velocity (SEV), saccade acceleration, saccade latency and self-rated sedation. Throughout the study daily symptom rating scales were kept. According to the daily symptom rating scales, patients were divided into two groups; one group who displayed a significant variance in negative mood symptoms during HRT (cyclicity) and one group with no cyclical changes in negative mood symptoms during treatment. During treatment with either E(2) alone or E(2)+progesterone the response in saccadic eye movement parameters and in self-rated sedation to pregnanolone was enhanced compared to pretreatment values. The SEV, saccade acceleration and sedation responses to pregnanolone was also increased in women expressing cyclicity in negative mood symptoms compared to women with no cyclical changes in negative mood during HRT. In conclusion, during treatment with either E(2) alone, or E(2)+progesterone, pregnanolone sensitivity was increased. Women expressing cyclicity in negative mood symptoms were more sensitive to pregnanolone than women without symptom cyclicity during HRT. PMID- 15358442 TI - The diurnal patterns of the adrenal steroids cortisol and dehydroepiandrosterone (DHEA) in relation to awakening. AB - The steroid hormones, cortisol and dehydroepiandrosterone (DHEA) are the two main peripheral secretory products of the hypothalamic-pituitary-adrenal stress neuroendocrine axis. The diurnal pattern of cortisol secretory activity has been well characterised. Various aspects of this pattern have been related to time of awakening, light exposure, psychological dimensions of affect, immune function and systemic health and well-being. DHEA is also an important adrenocortical steroid whose secretory activity has been related to immune function, psychological and health variables. The most pronounced feature of the diurnal cortisol cycle is a burst of secretory activity following awakening with a diurnal decline thereafter. We mapped DHEA secretory activity onto this cycle by measuring both steroids in saliva samples collected at distinct time points over the diurnal cycle, synchronised to awakening. Both steroids, particularly DHEA, showed stability across days of sample collection. A main distinction between cortisol and DHEA was that although DHEA was elevated in post-awakening samples compared with later in the day there was no evidence of an awakening stimulatory burst of DHEA secretory activity. Although DHEA in many respects paralleled cortisol secretory activity there was some dissociation; mean levels were positively but not tightly correlated. The secretory pattern of DHEA is very stable whereas cortisol secretory activity seems more sensitive to day-to-day variability. PMID- 15358443 TI - Gender differences in testosterone and cortisol response to competition. AB - This study examined intra-individual change in testosterone, cortisol, and hormone-behavior relationships in response to a rowing ergometer competition. Forty-six members (23 females) of a university crew team provided saliva samples before, 20- and 40-min post-competition, as well as baselines on a non competition day. Behavioral assessments included measures of previous rowing experience, dominance, competitiveness, bonding with teammates, pre- and post competition mental state and performance. Men's and women's endocrine responses to this competitive setting were more different than alike and varied by level of competitive experience, the specific phase of the competitive event, and the particular hormone measured. Inter-individual differences in testosterone and cortisol were differentially associated with social affiliation with teammates but rarely with dominance or competitiveness. Theoretically, the findings support the integration of features of the 'tend and befriend' model with the biosocial model of status, and suggest future research directions that may lead to clarification and refinement of those ideas. PMID- 15358444 TI - Twenty-four hour growth hormone secretion in patients with panic disorder. AB - BACKGROUND: Patients with panic disorder have blunted growth hormone (GH) responses to clonidine, suggesting subsensitivity of post-synaptic alpha(2) adrenoreceptors, presumably in response to excessive central noradrenergic outflow. However, basal levels of GH release over a full circadian cycle have not been examined in panic. Reduced basal GH release would suggest an overall hypo active GH system rather than a specific alpha-adrenergic abnormality. METHODS: To determine whether panic patients show reduced basal GH secretion, 20 patients and 12 healthy controls were studied. Blood samples were drawn every 15 min for 24 h and plasma was assayed for GH. Patients were restudied during successful treatment with alprazolam. Groups were compared on overnight and daytime GH secretion and circadian patterns of release. RESULTS: Patients showed normal levels on all measures of GH release. Treatment may have reduced nocturnal GH release slightly, but treated patients still did not differ from controls. The normal predominance of sleep over waking GH secretion was seen in both groups. CONCLUSIONS: Panic patients, in contrast to depressed patients, have normal somatotrophic axis activity when measured in a resting state over a full circadian cycle. GH dysregulation may only be evident in these patients in activation paradigms and has been most consistently demonstrated by challenges with the alpha(2)-noradrenergic agonist, clonidine. PMID- 15358445 TI - A meta-analysis of cortisol response to challenge in human aging: importance of gender. AB - An increased cortisol response to challenge is associated with a variety of age related disorders such as Alzheimer's disease, depression, diabetes, metabolic syndrome, and hypertension. Among the healthy elderly, an increased cortisol response to challenge may be a risk factor for developing these age-related disorders. We searched Pubmed, Embase, PsychInfo, Biosis, and Digital Dissertations (January 1966-June 2003) and included 45 parallel-group (young vs. old subjects) studies that used either a pharmacological or psychological challenge in healthy volunteers and measured cortisol response to challenge. We calculated effect sizes (Cohen's d) for the standardized mean differences between groups. Compared to younger controls (n=670, mean age 28 years +/-5), older subjects (n=625, 69+/-6) showed a larger cortisol response to challenge defined as stronger response to stimulation or less inhibition after a suppression test (d=0.42, 95% confidence interval (CI), 0.26-0.57). The effect of age on cortisol release was significantly stronger in women (d=0.65, 95% CI 0.34-0.97) than men (d=0.24, 95% CI 0.02-0.47). Our results demonstrate that aging increases the cortisol response to challenge. This effect of age on cortisol response is almost three-fold stronger in women than men. Prospective studies should explore whether the higher cortisol response in the elderly is a risk factor for developing neuropsychiatric and medical disorders. PMID- 15358446 TI - Diurnal cortisol rhythm and fatigue in breast cancer survivors. AB - Approximately 30% of breast cancer survivors report persistent fatigue of unknown origin. We have previously shown that cancer-related fatigue is associated with alterations in immunological parameters and serum cortisol levels in breast cancer survivors. The current study examined the diurnal rhythm of salivary cortisol in fatigued and non-fatigued breast cancer survivors. Salivary cortisol measures were obtained from breast cancer survivors with persistent fatigue (n=13) and a control group of non-fatigued survivors (n=16). Participants collected saliva samples upon awakening and at 1200, 1700, and 2200 h on two consecutive days. Diurnal cortisol slope for each day was determined by linear regression of log-transformed cortisol values on collection time and analyzed using multi-level modeling. Fatigued breast cancer survivors had a significantly flatter cortisol slope than non-fatigued survivors, with a less rapid decline in cortisol levels in the evening hours. At the individual patient level, survivors who reported the highest levels of fatigue also had the flattest cortisol slopes. Group differences remained significant in analyses controlling for demographic and medical factors, daily health behaviors, and other potential confounds (e.g. depressed mood, body mass index). Results suggest a subtle dysregulation in hypothalamic-pituitary-adrenal axis functioning in breast cancer survivors with persistent fatigue. PMID- 15358447 TI - No morning cortisol response in patients with severe global amnesia. AB - Activity of the hypothalamus pituitary adrenal (HPA) axis is characterized by a pronounced circadian rhythm. An acute increase in cortisol levels occurs after awakening in the morning with continuously declining levels over the course of the remaining day. The morning cortisol increase probably reflects an activational response of the HPA axis aimed at preparing the body for the day. Some studies found patterns of enhanced or blunted waking cortisol responses observed under chronic stress, burnout, or post traumatic stress disorder. The present study wanted to characterize the morning cortisol response and the circadian cortisol day profile in a sample of six male patients with severe amnesia due to hypoxia, herpes simplex encephalitis or closed head injury. Age and gender matched relatives or friends served as controls. Cortisol was measured from saliva samples collected at home on two consecutive days. The patients were woken up in the morning by their partners or caregivers. The morning cortisol increase typically observed in healthy subjects and also observed in the control group was absent in the amnesic patients. In contrast, a normal circadian day profile was found in the amnesic patients, with a pronounced circadian cortisol decrease. Further studies are needed to understand the neurological or psychological mechanisms leading to a missing morning cortisol response in amnesic patients. PMID- 15358448 TI - Olanzapine increases plasma ghrelin level in patients with schizophrenia. AB - OBJECTIVE: Increased appetite and weight gain are frequently reported in treatment with olanzapine. However, the mechanism behind this appetite gain remains unclear. Ghrelin is a newly discovered appetite-stimulating peptide that has a role in the regulation of feeding behavior. Ghrelin is synthesized principally in the stomach, and the concentration of circulating ghrelin is negatively correlated with leptin and body fat mass. To elucidate the mechanism of appetite and weight gain during olanzapine treatment, we investigated the circulating ghrelin levels. METHODS: Seven patients with schizophrenia were examined before and after 6-month administration of olanzapine. The concentrations of circulating ghrelin, leptin, glucose and lipid metabolic parameters were measured. RESULTS: Body fat percentage (P=0.0121) and serum leptin (P=0.0284) were significantly increased after 6-month administration of olanzapine. Both plasma total ghrelin (P=0.0188) and active ghrelin levels (P=0.0057) were significantly increased. Six of the seven patients reported increased appetite during olanzapine treatment. Other glucose and lipid parameters were not altered significantly. CONCLUSIONS: Although, the leptin level and body fat percentage were significantly increased, the concentration of circulating ghrelin was also significantly increased. Olanzapine may directly act on the secretion of ghrelin and induce appetite, resulting in weight gain. PMID- 15358449 TI - Sex hormones in psychotic men. AB - For women at risk to develop schizophrenia, estradiol has been postulated to constitute a protective factor. Women suffering from psychotic disorders have accordingly been found to exhibit lower estradiol levels than controls. Our aim was to study gonadal function in psychotic men to determine the gender specificity of these observations, as available data in men are more scarce and conflicting and largely disregarded estradiol. Serum hormone levels were examined in 34 men admitted consecutively for an acute exacerbation or first onset of schizophrenia in a blinded prospective design. Subjects with current affective disorder including manic episode, concomitant substance abuse or severe medical illness were excluded. A control group of 34 healthy male blood donors was recruited. As compared to matched controls, acutely admitted men suffering from schizophrenia exhibited significantly lower serum levels of estradiol, oestrone, testosterone and free testosterone. Although results have to be regarded as preliminary, acute exacerbation of schizophrenia in men seems to be associated with low serum oestrogen and androgen levels. The oestrogen hypothesis postulating a protective action of estradiol concerning schizophrenia for women might well be valid for both genders. However, future research is needed before clinical applications are justified. PMID- 15358450 TI - Acyl and desacyl ghrelin in anorexia nervosa. PMID- 15358452 TI - The obstetrician and depression during pregnancy. AB - The objective of this article is to review the literature as to the presence of depression during and after pregnancy and some of its clinical implications; and to present a simple statistical aide for screening purposes. Clinical depression affects at least one in five women of childbearing age. During pregnancy, this figure does not diminish and not only signals problems for the pregnant woman but also for the child, measurably so into adolescence. Postpartum depression, but even more so antepartum depression, are medical conditions that negatively affect mother and child, and need to be detected as early as possible to avoid or limit the use of pharmacological treatments with possible side effects. The obstetrician should regularly test for depression from the very first moments of planning for a child, and use the test results for a "pregnancy mood profile". This profile requires only a few minutes and is very simple to complex. It could serve for early control of depression during pregnancy as well as determine the risk for postpartum depression and thus serve as a pre-alert for postpartum suicide. PMID- 15358453 TI - Aneurysmal subarachnoid haemorrhage in pregnancy. AB - Although uncommon, aneurysmal subarachnoid haemorrhage (SAH) in pregnancy can be devastating for both mother and baby. It is the leading cause of other indirect maternal death in England and Wales accounting for 60 deaths in the decade 1988 1999. No single obstetric or neurosurgical unit has sufficiently large database or experience in managing this condition in pregnancy. With significant improvements in antenatal care and management of deliveries, non-obstetric causes of maternal death such as aneurysmal subarachnoid haemorrhage are likely to become increasingly significant. The clinical features of aneurysmal subarachnoid haemorrhage closely resemble those of other commoner conditions seen in pregnancy. It is therefore imperative that awareness by obstetricians and other frontline staff is increased so that a high index of suspicion is maintained when pregnant women present with unique headaches. Prompt neurosurgical referral is vital and early involvement of an experienced neuroradiologist essential. It is only when an early diagnosis is made and an aggressive treatment instituted that the bleak case-fatality figure associated with aneurysmal subarachnoid haemorrhage in pregnancy can be improved. This review, by a multidisciplinary and multicenter team, provides a comprehensive update on the epidemiology, aetiology, clinical presentation, diagnosis and the complexities of the multidisciplinary management of this serious and potentially fatal condition when it occurs in pregnancy. PMID- 15358454 TI - Female genital mutilation in developing countries: an agenda for public health response. AB - There is uncertainty regarding reasons for persistence of the practice of female genital mutilation (FGM) and the best strategies for intervening effectively. In spite of strong international condemnation, the persistence of FGM in many countries suggests that it can only be effectively eliminated when its practitioners are presented with a safe alternative that preserves their culture and, at the same time, protects the health and well being of women. Recognizing that there is no simple solution to the problem, this paper argues that interventions for preventing FGM should be non-directive, culture-specific and multi-faceted to be of practical relevance. Such interventions should not only motivate change, but should also help communities to establish practical means by which that change can occur. Potentially effective prevention interventions targeted at local practitioners of FGM, parents, at-risk adolescents, health and social workers, governments, religious authorities, the civil society, and communities are presented. PMID- 15358455 TI - Relationship between a toll-like receptor-4 gene polymorphism, bacterial vaginosis-related flora and vaginal cytokine responses in pregnant women. AB - OBJECTIVE: The relationship between a single nucleotide polymorphism (TLR4 896 A > G) in the toll-like receptor-4 (TLR4) gene, qualitative and quantitative changes in vaginal micro-flora and vaginal interleukin (IL)-1beta and IL-1 receptor antagonist (IL-1ra) concentrations in pregnant women were evaluated. STUDY DESIGN: Qualitative and quantitative microbial methods were used to characterize vaginal micro-flora of 238 women at 18-22 weeks gestation. Polymerase chain reaction was used to determine TLR4 genotype. IL-1beta and IL 1ra concentrations in vaginal lavage samples were measured by ELISA. RESULTS: The TLR4 variant was identified in 10.3% of women. Carriage of this variant was associated with a median increase in vaginal pH (P = 0.05), a greater than 10 fold increase in vaginal Gardnerella vaginalis levels (P < 0.0001) and a 10-fold increase in the vaginal concentration of three species of anaerobic Gram-negative rods, Prevotella, Bacteroides, and Porphyromonas (P = 0.08 ). Colonization with G. vaginalis and/or the anaerobic Gram-negative rods resulted in elevated vaginal IL-1 (P = 0.01) and IL-1ra (P < 0.0002) concentrations in women who were TLR4 896A homozygotes, but not in TLR4 896G carriers. CONCLUSION: The TLR4 896 A > G polymorphism contributes to inter-individual differences in the vaginal immune defense against G. vaginalis and anaerobic Gram-negative rods. PMID- 15358456 TI - Preterm birth reduction after clotrimazole treatment during pregnancy. AB - OBJECTIVE: We have previously found an association between the combination of topical and vaginal clotrimazole treatment during pregnancy and a decreased prevalence of preterm births in the population-based data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities. Thus the objective of this secondary analysis in the expanded data set was to evaluate potential confounders and to examine the possible interaction of clotrimazole with other drugs. STUDY DESIGN: Medically recorded birth weight/gestational age, in addition the prevalence of preterm birth and low birthweight infants of newborn infants without birth defects born to mothers with or without clotrimazole treatment during pregnancy were compared in the expanded control data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1996. RESULTS: The 17 year data set included 38,151 newborn infants and 8.1% were born to mothers who received clotrimazole treatment during pregnancy. There was an increase in mean gestational age among the exposed relative to the unexposed, resulting in a significant (34-64%) reduction in the prevalence of preterm births. This finding could not be explained by confounders and/or interaction with other drugs. CONCLUSION: The protective effect of clotrimazole for preterm birth was confirmed. We conclude that the protective effect of topical clotrimazole during pregnancy may be attributable to the beneficial effect of clotrimazole in the restoration of the abnormal colonization of the female genital organs and its known antibacterial and/or antiprotozoal effect. PMID- 15358457 TI - Effectiveness of detection of intrauterine growth retardation by abdominal palpation as screening test in a low risk population: an observational study. AB - OBJECTIVE: To evaluate the performance of abdominal palpation as a screening test for intrauterine growth retardation (IUGR) in a low risk population, under standard practice conditions. STUDY DESIGN: Population based observational study of 6318 consecutive low risk singleton pregnancies. The Dutch obstetric system distinguishes low from high risk pregnancies. In the low risk group abdominal palpation as a screening test is performed by midwives. If a complication, like IUGR, during prenatal care is assessed, the women is referred to a consulted obstetrician. Ultrasound is performed by the consulted obstetrician. In case of sustained suspicion the women is selected as high risk. OUTCOME PARAMETERS: severe small for gestational age (SGA) birthweight below 2.3rd centile, all SGA birthweight below 10th centile, operative delivery, neonatal morbidity and perinatal mortality. Screening value of abdominal palpation, abdominal palpation combined with ultrasound, and the performance of high risk selection was assessed by conventional performance measures. RESULTS: Abdominal palpation as a screening test for IUGR is of limited value: the observed sensitivities were 28% for severe SGA and 21% for SGA p < or = 10, respectively. After ultrasound in case of sustained suspicion, the sensitivity in detection of severe SGA was 25% and positive predictive value (PPV) 16%. In detection of SGA p < or = 10 sensitivity was 15% and PPV 55%, which means 45% were false positives. The sensitivity of the Dutch obstetric system in selection of high risk pregnancies in detection of severe SGA was 53%, in detection of SGA p < or = 10 was 37%. Perinatal mortality was 0.9% (57/6318) and 32% of these cases were SGA. Six cases of fetal death were unrecognised during prenatal care (0.09%) and seem preventable. The prevalence of a 5 min Apgar Score < or = 7 was significantly higher in the SGA infants if SGA was defined as p < or = 10. CONCLUSIONS: The diagnostic performance of abdominal palpation as a screening test for IUGR detection in a low risk population is disappointing. However, various stratagems such as routine ultrasound do not improve detection rate or perinatal morbidity and mortality. PMID- 15358458 TI - Delivery and immigration: the experience of an Italian hospital. AB - OBJECTIVE: We studied mode of delivery and prevalence of complications in pregnant women from the western world (WW) and immigrant mothers from non European Union (non-EU) countries at a third level Italian Obstetric Department. STUDY DESIGN: The study was population based and used data from the local Birth Registry at the University of Bologna. A 1:1 case control was performed by matching 510 single live births from immigrant mothers (non-EU) during the period 1997-2001 with 510 controls in chronological order (WW). Data were matched by age and parity. RESULTS: No differences between the two groups were noted as concerns preterm delivery, percentage of infants transferred to neonatal intensive care unit, perinatal mortality, caesarean section rate, episiotomies, instrumental deliveries and post-partum complications. Significant differences were noted in the rate of elective caesarean section (which was higher in the WW women: P < 0.01) and in the rate of vaginal lacerations and neonatal malformations (which was higher in the non-EU group: P < 0.05). CONCLUSIONS: There was no substantial variation in the mode of delivery between non-EU immigrants and western women; only the rate of elective caesarean section was significantly higher in the WW group. A higher rate of vaginal lacerations and neonatal malformations was found in the non-EU group. In our experience the standard of medical care is achievable regardless of ethnic group. PMID- 15358459 TI - Induction of labor with prostaglandin E2 in women with previous cesarean section and unfavorable cervix. AB - OBJECTIVE: To study the pregnancy outcome of induction of labor with prostaglandin E2 (PGE2) in women with one previous lower segment cesarean section. METHODS: A retrospective cohort design was used. The study sample included 1028 consecutive women with one previous cesarean section, of whom 97 underwent induction of labor (study group) and 931 were admitted with spontaneous onset of labor (control group). Vaginal tablets of PGE2 were used for cervical ripening in the study group. Mode of delivery, neonatal outcome, indications for cesarean section, and rate of uterine rupture were compared between the groups. RESULTS: There were no significant differences between the study and control groups in mean (+/-S.D.) maternal age (30.9 +/- 4.7 years versus 31.2 +/- 4.8 years, P = 0.6), gestational age at delivery (39.2 +/- 1.8 weeks versus 39.3 +/- 1.6 weeks, P = 0.36), overall rate of cesarean section (36% versus 37.3%, P = 0.8), rates of low 5-min Apgar score < or =7 (3.1% versus 3.7%, P = 0.67) or cesarean section performed for nonreassuring fetal heart rate (6.1% versus 3.1%, P = 0.1). There were four cases of uterine rupture, all in the control group compared to none in the study group (nonsignificant). CONCLUSION: The findings suggest that induction of labor in women with one previous cesarean section does not increase the risk of cesarean section rate and does not adversely affect immediate neonatal outcome. We cautiously suggest that when there is no absolute indication for repeated cesarean section, induction of labor may be considered. PMID- 15358460 TI - Procalcitonin in cervicovaginal secretion in pregnancies complicated by preterm labor--a preliminary report. AB - OBJECTIVES: Although the usefulness of procalcitonin (PCT) in clinical practice is increasing, no data are available on procalcitonin during pregnancy. The purpose of this study was to investigate whether procalcitonin was present in the cervicovaginal secretion of pregnant women and, if so, to evaluate the practical value of determining the concentration. STUDY DESIGN: A total of 53 patients in whom preterm labor (PTL) was expected and 31 healthy pregnant women were enrolled in this study. In the preterm labor group procalcitonin concentrations were analyzed with reference to laboratory indices suggestive of infection. The outcome of pregnancy was recorded in each case, with mention of the gestational age at delivery and of the time between admission to hospital and delivery. RESULTS: Concentrations of procalcitonin in the preterm labor group were comparable to those in the healthy pregnant women. In the preterm labor group no significant correlations were observed between procalcitonin concentration and laboratory indices of infection. Nor were any correlations observed between procalcitonin concentration at the onset of preterm labor and gestational age either at the onset of labor or at delivery. However, procalcitonin concentrations at the onset of preterm labor were higher in patients who delivered prematurely than in those who delivered near term after treatment to delay labor. Procalcitonin concentrations in women whose babies were delivered within 3, 7 and 14 days of admission and in those whose babies were born at later times were comparable. CONCLUSIONS: In this study, procalcitonin was determined in the cervicovaginal secretion of pregnant women for the first time. However, no association was observed either between procalcitonin concentration at the onset of preterm labor and laboratory signs of infection or between procalcitonin concentration and time between admission to hospital and delivery. Procalcitonin determination would be unsatisfactory as a prognostic indicator of the length of time between admission to hospital and delivery. PMID- 15358461 TI - Induction of labor in pregnancies with suspected large-for-gestational-age fetuses and unfavorable cervix. AB - OBJECTIVES: To evaluate our single-center, single-team experience with induction of labor in pregnancies with suspected large for gestational age (LGA) fetuses. STUDY DESIGN: A retrospective case-controlled design was used. Non-diabetic patients with a suspected LGA fetus (estimated fetal weight > or =90th percentile) (group 1, n = 135) were compared with healthy patients admitted for elective induction of labor for either post-date pregnancy or a subjective perception of decreased fetal movements in the presence of normal fetal heart monitoring and biophysical profile (group 2, n = 326), and with healthy women with normal pregnancies and spontaneous onset of labor (group 3, n = 574). RESULTS: There were no between-group differences in maternal age, parity, number of prostaglandin E2 (PGE2) tablets used, instrumental delivery rate or Apgar scores. The rate of cesarean section (CS) was significantly higher in the study group (33.3%) than in group 2 (17.8%, P = 0.001) and group 3 (10.6%, P = 0.004), although this difference disappeared when the multiparous women were analyzed separately (study group: n = 58, 10.3% versus group 2: n = 169, 7.7% and group 3: n = 308, 7.8%, P = 0.6). A logistic regression model (R2 = 0.385, P < 0.001) was used to control for maternal and gestational age, nulliparity rate, number of PGE2 tablets used, birth weight, and diagnosis (group 1 versus group 2) as predictors of mode of delivery. On stepwise (forward-likelihood) analysis, only nulliparity (odds ratio (OR) 10.0, 95% confidence interval (CI) 2.8-35.6, P < 0.001) and maternal age (OR 1.2, 95% CI 1.06-1.36, P = 0.002) were independently and significantly associated with increased risk of CS. CONCLUSIONS: Induction of labor for suspected LGA fetuses, if performed at all, should be reserved for multiparous women. PMID- 15358462 TI - Cytotoxin associated gene A-positive Helicobacter pylori strains in dyspeptic pregnant women. AB - OBJECTIVE: The aim of the present study was to compare the Helicobacter pylori (HP) seropositivity and cytotoxin associated gene A (cagA) status in pregnant women with dyspeptic complaints and pregnant women with no gastrointestinal symptoms. STUDY DESIGN: Seventy-one consecutive pregnant women with gastrointestinal complaints and 72 age-matched pregnant women without any gastrointestinal symptoms or a history of gastrointestinal disease were included in the study. Demographic characteristics and H. pylori and cytotoxin associated gene A status of the groups were analysed. RESULTS: The prevalence of H. pylori seropositivity was slightly but not significantly higher in patients with dyspeptic complaints compared to the controls (74.6% versus 63.8%, respectively, P > 0.05). The incidence of dyspeptic complaints were 53.5% in HP-seropositive and 40.9% in HP-seronegative women (P > 0.05). The prevalence of cytotoxin associated gene A positivity among H. pylori-seropositive women was significantly higher in dyspeptic pregnants compared to the controls (75.5% versus 45.7%, respectively, P = 0.002). Among HP-seropositive women, the incidence of dyspeptic complaints was significantly higher in cagA-positive patients compared to the cagA-negative ones (65.6% versus 34.2%, respectively, P = 0.002). When analysed according to the trimesters, the prevalence of cytotoxin associated gene A positivity among H. pylori-seropositive women was significantly higher in dyspeptic pregnants compared to the controls in the first trimester (68.0% versus 34.8%, respectively, P = 0.021). CONCLUSION: Cytotoxin associated gene A positive, virulent H. pylori strains were found to be more frequently associated with dyspeptic complaints in pregnant women. PMID- 15358463 TI - Attitudes and behavior towards contraception among Greek women during reproductive age: a country-wide survey. AB - OBJECTIVES: The study objectives were to explore current contraceptive behavior of Greek women during reproductive age. STUDY DESIGN: Data were drawn from a country-wide survey, conducted with the use of a self-administered questionnaire. The sample, numbering 797 women of ages 16-45 years, was representative of the Greek female population of reproductive age. RESULTS: The most common contraceptive method reported was the male condom (MC) (33.9%) followed by coitus interruptus (CI) (28.8%), oral pill (4.8%), and coil (3.6%). Other methods counted for a 5% and no use of contraception reached a 23.8%. Attitudes over responsibility of using contraception were also explored. The majority of respondents (52%) stated that contraception use is the responsibility of men. The probability of reporting that women should be responsible in using contraception was higher in women aged 25-34 years, in those with higher level of knowledge over contraception issues and in those with an experience of abortion. CONCLUSIONS: The need for sexual education and easy access to counseling services is apparent in order to promote optimal contraception decision-making. The role of women in taking active responsibility over contraception use should be of great importance in reproductive health promotion projects. PMID- 15358464 TI - A randomized control comparison study of culture media (HTF versus P1) for human in vitro fertilization. AB - OBJECTIVE: It is now widely accepted that increasing the number of replacement embryos (>3 embryos per embryo transfer [ET]) is associated with an increased risk of multiple pregnancies. While embryo reduction is often proposed when there is a high risk of multiple pregnancies, it is a difficult decision for the couple. For this reason, different studies have focused on single embryo transfer, more precisely blastocyst transfer. The aim of the study is to confirm that phosphate-free culture media can be used to generate greater quality embryos. METHODS AND RESULTS: We carried out a study to compare the efficacy of human tubal fluid (HTF) versus preimplantation stage one (P1) as culture media for assisted reproductive therapy (ART). In 109 nonselected patients, we obtained an embryo fertilization rate with HTF and P1 culture media of 58.6 and 62.5% (P = 0.003), respectively. After 48 and 72 h, the morphology was similar for both P1 and HTF embryos in most patients. However, in the same patients, when HTF embryo quality was low (15.4%), P1 embryo quality was significantly higher 68.7% (P = 0.002). Some embryos were transferred at 48 h and some at 72 h after retrieval, in a randomized manner. We transferred a maximum of up to three embryos per ET. The implantation rate was significantly different; at 48 h, it was 6.8 and 12.2% for HTF and P1, respectively (P = 0.02). The pregnancy rate was 17.1% for HTF embryos and 23.7% for P1 embryos (P = 0.02). CONCLUSIONS: Therefore, we observed a significant difference between P1 and HTF in the fertilization rate, in embryo quality, in implantation rate and in pregnancy rate. But the most important difference between this study and others is that every patient was the control of herself, so we eliminated every variable. PMID- 15358465 TI - Premenstrual syndrome and associated symptoms in adolescent girls. AB - OBJECTIVE: To investigate the frequency of premenstrual syndrome (PMS) associated symptoms and effects of nutrition on PMS in adolescent girls. PATIENTS AND METHODS: One hundred and seventy-one adolescent girls who had menstrual cycles were included in this study. They were given a questionnaire on criteria for PMS, dysmenorrhea and regularity of menstrual cycle. Modified Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV) criteria were used for the diagnosis of PMS. We also investigated which nutritional supplements affect the PMS-associated symptoms and signs. RESULTS: One hundred and five adolescent girls out of 171 (61.4%) met DSM-IV criteria for PMS. There was an association between dysmenorrhea and PMS in 60 (57.1%). Half of the girls, i.e. 52 (49.5%) had mild, 39 (37.1%) had moderate and 14 (13.4%) had severe PMS. The most common symptom of PMS was negative affect particularly in the form of stress (87.6%) and nervousness (87.6%). There was a statistically significant negative relationship between milk consumption and the following: abdominal bloating, cramps, craving for some foods and increased appetite. CONCLUSION: PMS and dysmenorrhea are frequently overlapping. We also found that PMS is associated with dietary habits. PMID- 15358466 TI - Adnexal torsion in very young girls: diagnostic pitfalls. AB - OBJECTIVE: Evaluation of cases of young girls diagnosed with adnexal torsion. STUDY DESIGN: Retrospective cohort study of 39 girls born between January 1980 and January 2000 who presented with acute abdominal pain and/or the diagnosis adnexal torsion in the Groene Hart Hospital. Seven patients, aged 6-13 years, with adnexal torsion were further evaluated. RESULTS: In three patients, it was possible to save the adnexa. Only the time relapse between the onset of complaints and the surgical intervention correlated with the presence of microscopic necrosis in the ovary. Bluish-black appearance of an ovary did not correlate with the presence of microscopic necrosis. CONCLUSIONS: Early recognition and treatment are essential to minimise the risk of decreased fertility after adnexal torsion in young girls. We therefore advocate prompt diagnostic laparoscopy and detorsion of the adnexa. In case a cyst is present, it should not be resected, but checked on a regular basis using ultrasound. PMID- 15358467 TI - Randomized, double-blind, controlled trial of mifepristone in capsule versus tablet form followed by misoprostol for early medical abortion. AB - OBJECTIVE: To compare the efficacy and side-effects of mifepristone 75 mg in capsule form versus 150 mg in tablet form followed by misoprostol for medical termination of early pregnancy. STUDY DESIGN: In a prospective randomized, double blind, placebo-controlled trial, a total of 480 women who were 49 days or less pregnant were randomized by means of a random number table to receive either two tablets in the morning and one tablet 12 h later for 2 days (group A) or three capsules orally twice daily for 2 days, the first dose being double all subsequent doses (group B). After a further 48 h, 600 microg misoprostol was given orally. Successful abortion was defined as complete abortion with no need for surgical aspiration. RESULTS: There were no significant differences between the two study groups in the rates of complete abortion (95.4% in group A versus 96.3% in group B), incomplete abortion (3.8% in group A, 3.3% in group B) and continued pregnancy (0.8% in group A, 0.4% in group B). No significant difference in the duration and amount of vaginal bleeding was observed. The incidence of side-effects, such as vomiting, nausea, headache, diarrhea and lower abdominal pain was similar in the two groups. CONCLUSIONS: Our results indicate that 75 mg mifepristone in capsule form combined with 600 microg misoprostol is as effective and safe as 150 mg mifepristone in tablet form for the termination of pregnancy up to 49 days. PMID- 15358468 TI - Vaginal myomectomy using posterior colpotomy: feasibility in normal practice. AB - BACKGROUND: Myomectomy is classically performed via laparotomy. Endoscopic surgery is limited for some indications. Vaginal myomectomy is a surgical procedure that has recently been evaluated. The aim of this study was to evaluate the feasibility, reproducibility and complication rate of vaginal myomectomy via posterior colpotomy. METHODS: A retrospective study was performed from November 1998 to February 2001 in three departments of gynaecology. RESULTS: Forty-five patients were involved, 17 (37.8%) of whom underwent laparoscopy before vaginal myomectomy for evaluation or treatment of a second pelvic disorder. Myomectomy was performed vaginally in 40 (89%) of the 45 patients. Peroperative laparotomy was required in five patients (11%), either because vaginal myomectomy proved impossible (four cases) or because of rectal injury (one case). One patient needed supplementary laparotomy on day 8 for the treatment of a pelvic abscess. CONCLUSION: Posterior vaginal myomectomy seems to be a feasible and reproducible surgical procedure. The success rate of vaginal myomectomy in this study was over 80%. Further randomised studies are needed to compare this procedure with laparotomy and laparoscopy. PMID- 15358469 TI - Medical treatment of ruptured with hemodynamically stable and unruptured ectopic pregnancy patients. AB - OBJECTIVE: To determine the success rate of methotrexate treatment of ruptured ectopic pregnancy with hemodynamically stable and unruptured ectopic pregnancy patients. STUDY DESIGN: This prospective clinical study was carried out on 161 patients with suspected tubal ectopic pregnancy. Fourty-six patients have been accepted as ruptured ectopic pregnancy with hemodynamically stable and 115 patients have been accepted as unruptured ectopic pregnancy. All patients diagnosed with ectopic pregnancy were treated by single dose (50 mg/m2) methotrexate if they have stable hemodynamia and fulfill the criteria of methotrexate treatment. Weekly beta-hCG level was measured and if this level was under 10 IU/L, the treatment has been accepted as successful. Mann-Whitney and Fisher's exact tests were used (SPSS, 10.0) for statistical analysis. RESULTS: The success rates of methotrexate treatments in ruptured ectopic pregnancy patients with hemodynamically stable and in patients with unruptured ectopic pregnancy were observed as 62% and 81%, respectively (P < 0.001). The treatment was successfully completed in all expectant management patients. CONCLUSION: Although methotrexate treatment of ruptured ectopic pregnancy with hemodynamically stable patients is not as successful as in unruptured ectopic pregnancy group, 62% success rate in this group may promise a treatment choice before surgery application. PMID- 15358470 TI - U-937 monocyte-mediated c-Jun dephosphorylation and AP-1 activation in human endometrial stromal cells. AB - OBJECTIVE: To determine paracrine effects of monocytes/macrophages on c-Jun dephosphorylation and AP-1 DNA binding activity in human endometrial stromal cells. STUDY DESIGN: Conditioned medium (CM) was prepared from human monocyte U 937 cells in serum-free medium. Subconfluent immortalized human endometrial stromal N5 cells were serum-starved for 24 h and cultured in the CM or the control medium for 30 min, 1, 3, 8, 16 or 24 h. Nuclear extracts were prepared and phosphorylated and dephosphorylated c-Jun isoforms were detected by Western blot analysis and the DNA binding activity was evaluated by electrophoretic mobility shift assay. Data on protein levels and DNA binding activity were analyzed statistically by ANOVA using SAS programs. RESULTS: c-Jun was dominantly in the phosphorylated state in N5 cells cultured in the control medium. The CM induced c-Jun accumulation and dephosphorylation and increased AP-1 DNA binding activity in a time-dependent manner. CONCLUSION: U-937 cells induce c-Jun dephosphorylation and AP-1 activation in human endometrial stromal cells by paracrine factors. Further investigations are needed to characterize the nature of these paracrine factors and their signaling pathways leading to AP-1 activation. PMID- 15358471 TI - Neonatal haemochromatosis: a rare cause of neonatal death. AB - Neonatal haemochromatosis is a very rare disorder with an unclear aetiology and an extremely poor outcome. A 24 year-old patient was delivered at 32 week gestation. The new-born presented a liver failure syndrome and died shortly after birth. The diagnosis of haemochromatosis was based on iron deposition revealed by the autopsy. In families at risk, ultrasound foetal follow up should be proposed and the detection of an anomaly suggestive of NH recurrence justifies a MRI in early third trimester. Liver transplantation is the treatment of choice for advanced disease. PMID- 15358472 TI - Pregnancy and delivery with an abdominal mesh graft. AB - Pregnancy and delivery were uneventful in a woman who conceived within six weeks after an abdominal reconstruction with a Marlex mesh graft for aesthetic reasons. One year after vaginal delivery, the abdominal wall reconstruction was still completely intact. PMID- 15358473 TI - Factor VII deficiency and its treatment in delivery with recombinant factor VII. AB - The authors present two cases of pregnant women with coagulation disorders--one with inherited deficiency of factor VII and the second with chronic hepatitis. Both cases were successfully treated with recombinant activated factor VII of coagulation (NovoSeven) in delivery. The advantages of using this product are discussed. PMID- 15358474 TI - Comment on "Anterior vaginal wall repair using local anaesthesia" [Eur. J. Obstet. Gynecol. Reprod. Biol. 112 (2004) 214-216]. PMID- 15358477 TI - Comment on "Clinical observation of blood loss reduced by tranexamic acid during and after cesarian section: a multi-center, randomized trial (Eur J Obstet Gynecol Reprod Biol 2004; 112: 154-57)". PMID- 15358478 TI - A patient with granulocyte-colony stimulating factor-producing endometrial cancer who responded to high-dose cisplatin, cyclophosphamide and adriamycin. PMID- 15358479 TI - Neonatal chronic renal failure associated with maternal ingestion of Nimesulide as analgesic. PMID- 15358480 TI - Multicystic peritoneal mesothelioma of the omentum. PMID- 15358481 TI - Pelvic Schwannoma. PMID- 15358482 TI - Focal cervical and vaginal necrosis following uterine artery embolisation. PMID- 15358484 TI - Transport of lead and diesel fuel through a peat soil near Juneau, AK: a pilot study. AB - A set of peat column experiments was used to determine the transport potential of lead (Pb) and diesel range organics (DRO) in palustrine slope wetlands near Juneau, AK. This project is important to southeast Alaskan communities because limited land resources are forcing development of regional wetlands. This study was instigated by concerns that proposed modifications to a nearby rifle range using DRO-contaminated soil posed a potential risk to an anadromous fish-bearing stream 250 m from the site. Three pairs of peat columns were extracted from the rifle range for analysis, one pair along and two pairs across the natural bedding planes of the soil. One column in each pair was spiked with Pb and DRO and the other was used as a control. Approximately 1-year worth of water (171 cm) was passed through each column and leachate was collected at regular intervals. The results showed that substantial DRO transport only occurred along the bedding planes. Leads was surprisingly mobile, both along and across the bedding planes with estimated soil-water partition coefficients several orders of magnitude lower than commonly published values, probably because the peat was heavily Pb loaded by lead from bullets and because the peat's acidic, organic-rich environment enhanced Pb mobility. The chemical outflow behavior agreed with a simple macropore transport model. These results underscore the need for caution when developing regional wetlands. PMID- 15358485 TI - The influence of system complexity on bacterial transport in saturated porous media. AB - A series of miscible-displacement column experiments were conducted under saturated flow conditions to systematically investigate the influence of physical and biological complexity on bacterial activity and fate in the presence and absence of a non-sorbing growth substrate, salicylate. Bacterial elution was monitored for three different systems; System I--a sterilized, inoculated, well sorted sand, System II--a sterilized, inoculated, heterogeneous loamy sand (Hayhook), and System III--two different unsterilized loamy sands (Hayhook and Vinton) each with their associated indigenous microbial community. Results show that System I behaved ideally with respect to both cell and substrate transport, wherein: (1) growth occurred in response to substrate addition, (2) cell elution increased in response to the substrate pulse, and (3) breakthrough curves were reproducible for both substrate and cell elution. In contrast, System II showed ideal behavior with respect to substrate transport but showed variable behavior for cell transport. Further, there was no measurable growth in response to substrate addition and no increase in cell elution during the salicylate pulse. System III exhibited non-ideal behavior for both substrate and cell transport. Of particular interest is the fact that the indigenous communities of the two soils behaved differently. Specifically, for the Hayhook soil, an increased elution response was observed for the heterotrophic population while the salicylate degrading community was preferentially retained in the column. In contrast for the Vinton soil, the substrate pulse did not elicit an elution response from either the heterotrophic or salicylate-degrading community from the culturable, indigenous Vinton microorganisms. For Systems II and III, the observed variability appears to be associated with the biological component of the system, since sterile controls were reproducible. This type of systematic study is critical for understanding cell and substrate transport behavior in complex, heterogeneous systems, and illustrates the potential uncertainty associated with measurements in such systems. PMID- 15358486 TI - Field investigation into unsaturated flow and transport in a fault: model analyses. AB - Results of a fault test performed in the unsaturated zone of Yucca Mountain, Nevada, were analyzed using a three-dimensional numerical model. The fault was explicitly represented as a discrete feature and the surrounding rock was treated as a dual-continuum (fracture-matrix) system. Model calibration against seepage and water-travel-velocity data suggests that lithophysal cavities connected to fractures can considerably enhance the effective fracture porosity and therefore retard water flow in fractures. Comparisons between simulation results and tracer concentration data also indicate that matrix diffusion is an important mechanism for solute transport in unsaturated fractured rock. We found that an increased fault-matrix and fracture-matrix interface areas were needed to match the observed tracer data, which is consistent with previous studies. The study results suggest that the current site-scale model for the unsaturated zone of Yucca Mountain may underestimate radionuclide transport time within the unsaturated zone, because an increased fracture-matrix interface area and the increased effective fracture porosity arising from lithophysal cavities are not considered in the current site-scale model. PMID- 15358487 TI - Secondary imbibition in NAPL-invaded mixed-wet sediments. AB - A previously developed pore network model is used here to study the spontaneous and forced secondary imbibition of a NAPL-invaded sediment, as in the displacement of NAPL by waterflooding a mixed-wet soil. We use a 3D disordered pore network with a realistic representation of pore geometry and connectivity, and a quasi-static displacement model that fully describes the pore-scale physics. After primary drainage (NAPL displacing water) up to a maximum capillary pressure, we simulate secondary imbibition (water displacing NAPL). We conduct a parametric study of imbibition by varying systematically the controlling parameters: the advancing contact angles, the fraction of NAPL-wet pores, the interfacial tension, and the initial water saturation. Once the secondary imbibition is completed, the controlling displacement mechanisms, capillary pressures, relative permeabilities, and trapped NAPL saturations are reported. It is assumed that NAPL migrates into an initially strongly water-wet sediment, i.e., the receding contact angles are very small. However, depending on the surface mineralogy and chemical compositions of the immiscible fluid phases, the wettability of pore interiors is altered while the neighborhoods of pore corners remain strongly water-wet-resulting in a mixed-wet sediment. Here, we compare three different levels of wettability alteration: water-wet (advancing contact angles (20 degrees to 55 degrees), intermediate-wet (55 degrees to 120 degrees), and NAPL-wet (120 degrees to 155 degrees). The range of advancing contact angles and the fraction of NAPL-wet pores have dramatic effects on the NAPL-water capillary pressures and relative permeabilities. The spatially inhomogeneous interfacial tension has a minor impact on the trapped NAPL saturation and relative permeability to NAPL, and a slight effect on the relative permeability to water. The initial water saturation has a slight effect on the two-phase flow characteristics of water-wet sediments; however, with more NAPL-wet pores in the sediment, it starts to have a profound effect on the water and NAPL relative permeabilities. PMID- 15358488 TI - Nonideal transport of reactive contaminants in heterogeneous porous media: 7. distributed-domain model incorporating immiscible-liquid dissolution and rate limited sorption/desorption. AB - The purpose of this work is to present a distributed-domain mathematical model incorporating the primary mass-transfer processes that mediate the transport of immiscible organic liquid constituents in water-saturated, locally heterogeneous porous media. Specifically, the impact of grain/pore-scale heterogeneity on immiscible-liquid dissolution and sorption/desorption is represented in the model by describing the system as comprising a continuous distribution of mass-transfer domains. With this conceptualization, the distributions of the initial dissolution rate coefficient and the sorption/desorption rate coefficient are represented as probability density functions. Several sets of numerical experiments are conducted to examine the effects of heterogeneous dissolution and sorption/desorption on contaminant transport and elution. Four scenarios with different combinations of uniform/heterogeneous rate-limited dissolution and uniform/heterogeneous rate-limited sorption/desorption are evaluated. The results show that both heterogeneous rate-limited sorption/desorption and heterogeneous rate-limited dissolution can significantly increase the time or pore volumes required to elute immiscible-liquid constituents from a contaminated porous medium. However, sorption/desorption has minimal influence on elution behavior until essentially all of the immiscible liquid has been removed. For typical immiscible-liquid constituents that have relatively low sorption, the asymptotic elution tailing produced by heterogeneous rate-limited sorption/desorption begins at effluent concentrations that are several orders of magnitude below the initial steady-state concentrations associated with dissolution of the immiscible liquid. Conversely, the enhanced elution tailing associated with heterogeneous rate limited dissolution begins at concentrations that are approximately one-tenth of the initial steady-state concentrations. Hence, dissolution may generally control elution behavior of immiscible-liquid constituents in cases wherein grain/pore scale heterogeneity significantly influences both dissolution and sorption/desorption. PMID- 15358489 TI - Refinement of the density-modified displacement method for efficient treatment of tetrachloroethene source zones. AB - A novel method to remediate dense nonaqueous phase liquid (DNAPL) source zones that incorporates in situ density conversion of DNAPL via alcohol partitioning followed by displacement with a low interfacial tension (IFT) surfactant flood has been developed. Previous studies demonstrated the ability of the density modified displacement (DMD) method to recover chlorobenzene (CB) and trichloroethene (TCE) from heterogeneous porous media without downward migration of the dissolved plume or free product. However, the extent of alcohol (n butanol) partitioning required for in situ density conversion of high-density NAPLs, such as tetrachloroethene (PCE), could limit the utility of the DMD method. Hence, the objective of this study was to compare the efficacy of two n butanol delivery approaches: an aqueous solution of 6% (wt) n-butanol and a surfactant-stabilized macroemulsion containing 15% (vol) n-butanol in water, to achieve density reduction of PCE-NAPL in two-dimensional (2-D) aquifer cells. Results of liquid-liquid equilibrium studies indicated that density conversion of PCE relative to water occurred at an n-butanol mole fraction of 0.56, equivalent to approximately 5 ml n-butanol per 1 ml of PCE when in equilibrium with an aqueous solution. In 2-D aquifer cell studies, density conversion of PCE was realized using both n-butanol preflood solutions, with effluent NAPL samples exhibiting density reductions ranging from 0.51 to 0.70 g/ml. Although the overall PCE mass recoveries were similar (91% and 93%) regardless of the n butanol delivery method, the surfactant-stabilized macroemulsion preflood removed approximately 50% of the PCE mass. In addition, only 1.2 pore volumes of the macroemulsion solution were required to achieve in situ density conversion of PCE, compared to 6.4 pore volumes of the 6% (wt) n-butanol solution. These findings demonstrate that use of the DMD method with a surfactant-stabilized macroemulsion containing n-butanol holds promise as an effective source zone remediation technology, allowing for efficient recovery of PCE-DNAPL while mitigating downward migration of the dissolved plume and free product. PMID- 15358490 TI - Batch-test study on the dechlorination of 1,1,1-trichloroethane in contaminated aquifer material by zero-valent iron. AB - Chlorinated aliphatic hydrocarbons are common groundwater contaminants. One possible remediation option is in-situ reductive dechlorination by zero-valent iron, either by direct injection or as reactive barriers. Chlorinated ethenes (tetrachloroethene: PCE; trichloroethene: TCE) have received extensive attention in this context. However, another common groundwater pollutant, 1,1,1 trichlorethane (TCA), has attracted much less attention. We studied TCA reduction by three types of granular zero-valent irons in a series of batch experiments using polluted groundwater, with and without added aquifer material. Two types of iron were able to reduce TCA completely with no daughter product concentration increases (1,1-dichloroethane: DCA; chloroethane: CA). One type of iron showed slower reduction, with intermediate rise of DCA and CA concentrations. When evaluating the formation of daughter products, the tests on the groundwater alone showed different results than the groundwater plus aquifer batches: DCA did not temporarily accumulate in the batches with added aquifer material, contrary to the batches without added aquifer material. 1,1-dichloroethene (DCE, also present in the groundwater as an abiotic degradation product of TCA) was also reduced slower in the batches without added aquifer material than in the batches with aquifer material. Redox potentials gradually decreased to low values in batches with aquifer material without iron, while the batches with groundwater alone maintained a constant higher redox potential. Either adsorption processes or microbiological activity in the samples could explain these phenomena. Polymerase Chain Reaction (PCR: a targeted gene probe technique) for chlorinated aliphatic compound (CAH)-degrading bacteria confirmed the presence of Dehalococcoides sp. (chloroethene-degraders) but was negative for Desulfobacterium autotrophicum (a known co-metabolic TCA degrader). DCA reduction was rate determining: first-order half-lives of 300-350 h were observed. TCA was fully removed within hours. CA is resistant to reduction by zero-valent iron but it is known to hydrolyze easily. Since CA did not accumulate in our batches, it may have disappeared by the latter mechanism or it may not have formed as a major daughter product. PMID- 15358491 TI - Laboratory evidence of natural remobilization of multicomponent DNAPL pools due to dissolution. AB - Mixtures of dense non-aqueous phase liquids (DNAPLs) trapped in the subsurface can act as long-term sources of contamination by dissolving into flowing groundwater. In general, the components of higher solubility are removed more quickly, thus altering the composition of the remaining DNAPL, and possibly leading to changes in its physical properties. Through the development of a simple compositional model, Roy et al. [J. Contam. Hydrol. 2002 (59) 163] showed that preferential dissolution of a mixed DNAPL could potentially result in changes in density and interfacial tension that could subsequently lead to remobilization of an initially static DNAPL pool. The laboratory experiments presented in this next paper provide a proof-of-concept for the previously presented theory, demonstrating and quantifying this process of remobilization. In addition, the experiments provide a data set for evaluation of the model presented by Roy et al. [J. Contam. Hydrol. 2002 (59) 163]. In the four experiments, a DNAPL pool comprised of tetrachloroethene and benzene was created as an open pool overlying glass beads within a water-saturated 2-D flow box. Experiments included rectangular and triangular pools. In each of the experiments, remobilization (as breakthrough) was observed more than 2 weeks after formation of the initial pool. During each experiment, the pool height declined as mass was lost by dissolution, while sampling indicated a decrease in the mole fraction of benzene, the more soluble component. Small protuberances formed along the bottom of the pool as its composition changed with time and the displacement pressure was achieved for various pore throats. Eventually one of the protuberances extended further, forming a finger (breakthrough). In general, the pool emptied as the finger proceeded further into the beads. It was also shown theoretically and experimentally that remobilization will occur sooner for pools with a triangular (pointing down), rather than rectangular, shape. The experimental results were simulated using the model developed by Roy et al. [J. Contam. Hydrol. 2002 (59) 163]. The model matched the observations well, suggesting that it accurately represents the primary mechanisms involved with natural remobilization under the conditions of the study. PMID- 15358492 TI - Assessment of interfacial mass transfer in water-unsaturated soils during vapor extraction. AB - This paper presents results of a numerical investigation of soil vapor extraction (SVE) systems at the laboratory scale. The SVE technique is used to remove volatile chlorinated hydrocarbons (VCHC) from the water-unsaturated soil zone. The developed numerical model solves equations of flow, transport and interfacial mass transfer regarding an isothermal n-component and three-phase system. The mathematical model is based on a simple pore network and phase distribution model and designed to be scaled by a characteristic length. All mathematical expressions are structured into VCHC specific and VCHC non-specific parameters. Furthermore, indicators are introduced that help to separate thermodynamic equilibrium from thermodynamic non-equilibrium domains and to determine the controlling physical parameters. For numerical solution, the system of partial differential equations is discretized by a finite volume method and an implicit Euler time stepping scheme. Computational effort is reduced notably through techniques that enable spatial and temporal adaptivity, through a standard multigrid method as well as through a problem-oriented sparse-matrix storage concept. Computations are carried out in two dimensions regarding the laboratory experiment of Fischer et al. [Water Resour. Res. 32 (12) 1996 3413]. By varying the characteristic length scale of the pore network and phase distribution model, it is shown that the experimental gas phase concentrations cannot be explained only by the volatility and diffusivity of the VCHC. The computational results suggest a sorption process whose significance grows with the aqueous activity of the less or non-polar organic compounds. PMID- 15358493 TI - Field study of TCE diffusion profiles below DNAPL to assess aquitard integrity. AB - An area where a free-product accumulation of trichloroethylene (TCE) dense non aqueous phase liquid (DNAPL) occurs at the bottom of a 10-m-thick surficial sand aquifer was studied to determine the integrity of the underlying, 20-m-thick, clayey silt aquitard formed of glaciolacustrine sediment. TCE concentration versus-depth profiles determined from aquitard cores collected at five locations indicated penetration of detectable TCE 2.5 to 3.0 m into the aquitard. Two of the profiles show persistent DNAPL at the aquitard interface, while two others indicate that DNAPL, present initially, was completely dissolved away producing concentration declines at the aquitard interface. The fifth profile suggests shallow DNAPL penetration (<0.5 m) into the aquitard, however, this penetration, which was likely caused by cross-contamination during core collection or cone penetrometry (CPT) of the aquitard interface, did not increase the maximum depth of TCE penetration. Combining the field profiles with one-dimensional model simulations, downward migration of the aqueous TCE front, defined as the EPA MCL of 5 microg/l, which was below the analytical detection limit, was projected to a distance between 4 and 5 m below the top of the aquitard. Using a single set of estimated aquitard parameter values, simulations of aqueous TCE migration into the aquitard provided a good fit to four of the field profiles with a migration time of 35 to 45 years, consistent with the history of TCE use at the site. These simulations indicate aqueous TCE migration is diffusion-dominated with only small advective influence by the downward groundwater velocity of 2 to 3 cm/year or less in the aquitard due to pumping of the underlying aquifer to supply water to the facility in the past 50 years. The applicability of the parameter values was confirmed by in situ diffusion experiments of 1-year duration, in which stainless steel cylinders containing DNAPL were inserted into the aquitard. The diffusion dominated nature of the profiles indicates that the aquitard provides long-term protection of the underlying aquifer from contamination from this DNAPL zone. Simulations of long-term migration of the TCE solute front indicate breakthrough to the lower aquifer at 1200 years for the no advection scenario and at 500 years if the strong downward hydraulic gradient persists. However, even after breakthrough, the mass flux through the aquitard to the underlying aquifer remains relatively low, and when considered in terms of potential impacts to pumping wells, concentrations are not expected to increase significantly above present-day MCLs. The use of contaminant profiles of different time and distance scales, in addition to hydraulic data, dramatically improves the ability to assess aquitard integrity, and provides improved transport parameter values for estimating contaminant arrival times and fluxes. The apparent lack of deep preferential pathways for TCE migration, such as open fractures, is probably due to the softness of the silty aquitard deposit and minimal physical or chemical weathering of the aquitard provides long-term protection of the underlying aquifer from contamination from this DNAPL zone. Simulations of long-term migration of the TCE solute front indicate breakthrough to the lower aquifer at 1200 years for the no advection scenario and at 500 years if the strong downward hydraulic gradient persists. However, even after the breakthrough, the mass flux through the aquitard to the underlying aquifer remains relatively low, and when considered in terms of potential impacts to pumping wells , concentrations are not expected to increase significantly above present-day MCLs. The use of contaminant profiles of different time and distance scales, in addition to hydraulic data, dramatically improves the ability to assess aquitard integrity, and provides improved transport parameter values for estimating contaminant arrival times and fluxes. The apparent lack of deep preferential pathways for TCE migration, such as open fractures, is probably due to the softness of the silty aquitard deposit and minimal physical or chemical weathering of the aquitard. PMID- 15358494 TI - Bacteriophage transport through a fining-upwards sedimentary sequence: laboratory experiments and simulation. AB - A column containing four concentric layers of progressively finer-grained glass beads (graded column) was used to study the transport of the bacteriophage T7 in water flowing parallel to layering through a fining-upwards (FU) sedimentary structure. By passing a pulse of T7, and a conservative solute tracer upwards through a column packed with a single bead size (uniform column), the capacity of each bead type to attenuate the bacteriophage was determined. Solute and bacteriophage responses were modelled using an analytical solution to the advection-dispersion equation, with first-order kinetic deposition simulating bacteriophage attenuation. Resulting deposition constants for different flow velocities indicated that filtration theory-determined values differed from experimentally determined values by less than 10%. In contrast, the responses of solute and bacteriophage tracers passing upwards through graded columns could not be reproduced with a single analytical solution. However, a flux-weighted summation of four one-dimensional advective-dispersive analytical terms approximated solute breakthrough curves. The prolonged tailing observed in the resulting curve resembled that typically generated from field-based tracer test data, reflecting the potential importance of textural heterogeneity in the transport of dissolved substances in groundwater. Moreover, bacteriophage deposition terms, determined from filtration theory, reproduced the T7 breakthrough curve once desorption and inactivation on grain surfaces were incorporated. To evaluate the effect of FU sequences on mass transport processes in more detail, bacteriophage passage through sequences resembling those sampled from a FU bed in a fluvioglacial gravel pit were carried out using an analogous approach to that employed in the laboratory. Both solute and bacteriophage breakthrough responses resembled those generated from field-based test data and in the graded column experiments. Comparisons with the results of simulations using averaged hydraulic conductivities show that simulations employing averaged parameters overestimate bacteriophage travel times and underestimate masses recovered and peak concentrations. PMID- 15358495 TI - Three-dimensional diffusion of non-sorbing species in porous sandstone: computer simulation based on X-ray microtomography using synchrotron radiation. AB - The diffusion pathways of porous sandstone were examined by a three-dimensional (3-D) imaging technique based on X-ray computed tomography (CT) using the SPring 8 (Super Photon ring-8 GeV, Hyogo, Japan) synchrotron radiation facility. The analysis was undertaken to develop better understanding of the diffusion pathways in natural rock as a key factor in clarifying the detailed mechanism of the diffusion of radionuclides and water molecules through the pore spaces of natural barriers in underground nuclear waste disposal facilities. A cylindrical sample (diameter 4 mm, length 6 mm) of sandstone (porosity 0.14) was imaged to obtain a 3-D image set of 450(3) voxels=2.62(3) mm(3). Through cluster-labeling analysis of the 3-D image set, it was revealed that 89% of the pore space forms a single large pore-cluster responsible for macroscopic diffusive transport, while only 11% of the pore space is made up of isolated pores that are not involved in long range diffusive transport. Computer simulations of the 3-D diffusion of non sorbing random walkers in the largest pore cluster were performed to calculate the surface-to-volume ratio of the pore, tortuosity (diffusion coefficient in free space divided by that in porous rock). The results showed that (i) the simulated surface-to-volume ratio is about 60% of the results obtained by conventional pulsed-field-gradient proton nuclear magnetic resonance (NMR) laboratory experiments and (ii) the simulated tortuosity is five to seven times larger than the results of laboratory diffusion experiments using non-sorbing I( ) and Br(-). These discrepancies are probably attributed to the intrinsic sample heterogeneity and limited spatial resolution of the CT system. The permeability was also estimated based on the NMR diffusometry theory using the results of the random walk simulations via the Kozeny-Carman equation. The estimated permeability involved an error of about 20% compared with the permeability measured by the conventional method, suggesting that the diffusometry-based NMR well logging with gradient coils is applicable to the in-situ permeability measurement of strata. The present study demonstrated that X-ray CT using synchrotron radiation is a powerful tool for obtaining 3-D pore structure images without the beam-hardening artifacts inevitable in conventional CT using X-ray tubes. PMID- 15358496 TI - Effect of source variability and transport processes on carbon isotope ratios of TCE and PCE in two sandy aquifers. AB - Chlorinated ethenes often migrate over extended distances in aquifers and may originate from different sources. The aim of this study was to determine whether stable carbon isotope ratios remain constant during dissolution and transport of chlorinated ethenes and whether the ratios can be used to link plumes to their sources. Detailed depth-discrete delineation of the carbon isotope ratio in a tetrachloroethene (PCE) plume and in a trichloroethene (TCE) plume was done along cross-sections orthogonal to groundwater flow in two sandy aquifers in the Province of Ontario, Canada. At the TCE site, TCE concentrations up to solubility were measured in one high concentration zone close to the bottom of the aquifer from where dense non-aqueous phase liquid (DNAPL) was collected. A laboratory experiment using the DNAPL indicated that only very small carbon isotope fractionation occurs during dissolution of TCE (0.26 per thousand), which is consistent with field observations. At most sampling points, the delta(13)C of dissolved TCE was similar to that of the DNAPL except for a few sampling points at the bottom of the aquifer close to the underlying aquitard. At these points, a (13)C enrichment of up to 2.4 per thousand was observed, which was likely due to biodegradation and possibly preferential diffusion of TCE with (12)C into the aquitard. In contrast to the TCE site, several distinct zones of high concentration were observed at the PCE site and from zones to zone, the delta(13)C values varied substantially from -24.3 per thousand to -33.6 per thousand. Comparison of the delta(13)C values in the high concentration zones made it possible to divide the plume in the three different domains, each probably representing a different episode and location of DNAPL release. The three different zones could still be distinguished 220 m from the DNAPL sources. This demonstrates that carbon isotope ratios can be used to differentiate between different zones in chlorinated ethene plumes and to link plume zones to their sources. In addition, subtle variations in delta(13)C at plume fringes provided insight into mechanisms of plume spreading in transverse vertical direction. These variations were identified because of the high-resolution provided by the monitoring network. PMID- 15358497 TI - The role of interfacial films in the mass transfer of naphthalene from creosotes to water. AB - Viscous, semi-rigid interfacial films that are formed at the interface of certain multi-component non-aqueous phase liquid (NAPLs) and water can significantly reduce the rates of mass transfer of solutes. Creosote-water systems were investigated for their ability to form interfacial films. The effects of these films on the creosote-water partition and on mass transfer of a representative solute, naphthalene, were investigated in a series of experiments. The area independent mass transfer coefficient of naphthalene contained in creosote decreased by 30% over a 1-week period in systems containing creosote and water. Further aging for up to 21 days did not result in significant additional decreases in the mass transfer coefficient. The creosote-water partition coefficient, however, did not change with extended contact. The presence of viscous interfacial films in creosote-water systems was demonstrated in pendant drop tests. These interfacial films most likely caused the reduction in solute mass transfer coefficients by providing significant resistance to the diffusion of solutes through the interfacial film. Results from mass transfer experiments conducted under different system conditions suggested that hindered diffusion of naphthalene through the bulk creosote phase, changes in composition of creosote as a result of extended dissolution, or changes in creosote-water interfacial area did not contribute to the decrease in naphthalene mass transfer coefficient. PMID- 15358498 TI - The influence of high initial concentration aqueous-phase TCE on the performance of iron wall systems. AB - Flow-through column tests were conducted to investigate the performance of iron wall remediation systems for the degradation of aqueous-phase trichloroethylene (TCE). Concentration profiles under steady-state transport conditions were generated by measuring TCE concentrations at sample ports located at various locations along the length of the column. The results indicated that a pseudo first-order model is adequate at describing degradation kinetics for low initial TCE concentrations, but not for higher initial concentrations. The deviation from pseudo-first-order kinetics can be explained by interspecies competition for reactive sites between TCE and a dominant reaction product. A modification of the pseudo-first-order model that accounts for product interference predicts laboratory data for high initial concentration profiles, but deviates slightly as initial concentrations approach the solubility of TCE. The data clearly demonstrate the importance of accurately describing reaction kinetics for the purpose of designing iron wall treatment systems. PMID- 15358499 TI - Simulated and experimental evaluation of factors affecting the rate and extent of reductive dehalogenation of chloroethenes with glucose. AB - Carbohydrates such as molasses are being added to aquifers to serve as electron donors for reductive dehalogenation of chloroethenes. Glucose, as a model carbohydrate, was studied to better understand the processes involved and to evaluate the effectiveness for dehalogenation of different approaches for carbohydrate addition. A simulation model was developed and calibrated with experimental data for the reductive dehalogenation of tetrachloroethene to ethene via cis-1,2-dichloroethene. The model included fermentors that convert the primary donor (glucose) into butyrate, acetate and hydrogen, methanogens, and two separate dehalogenator groups. The dehalogenation groups use the hydrogen intermediate as an electron donor and the different haloethenes as electron acceptors through competitive inhibition. Model simulations suggest first that the initial relative population size of dehalogenators and H(2)-utilizing methanogens greatly affects the degree of dehalogenation achieved. Second, the growth and decay of biomass from soluble carbohydrate plays a significant role in reductive dehalogenation. Finally, the carbohydrate delivery strategies used (periodic versus batch addition and the time interval between periodic addition) greatly affect the degree of dehalogenation that can be obtained with a given amount of added carbohydrate. PMID- 15358500 TI - Antagonistic activity against Helicobacter pylori infection in vitro by a strain of Enterococcus faecium TM39. AB - Lactic acid bacteria (LAB) strains from infant feces were screened for anti Helicobacter pylori use. In the beginning, we selected the strains based on their capability to adhere to the human intestinal epithelial cell (Int-407), colonial enterocyte-like Caco-2 cell, human cervical epithelioid carcinoma cell (HeLa), and human gastric carcinoma cell (TSGH 9201). Then, acid and bile salt tolerance of these LAB strains was evaluated. In addition, the ability of these LAB strains to inhibit the growth of H. pylori and to expel H. pylori cells from TSGH 9201 were studied. The spent culture supernatant (SCS) of a selected strain TM39, i.e., TM39-SCS, significantly inhibited the viability of H. pylori in vitro. It also inhibited the urease activity of H. pylori in vitro. For these antagonistic effects, in addition to pH and lactic acid, some factors in TM39-SCS might play the major role. Treatment of H. pylori with the SCS or cells of strain TM39 significant reduced its binding to TSGH 9201 cells. Although strain TM39 is identified as Enterococcus faecium, it is not vancomycin resistant and is proved to be safe through the invasion study and a 28-day feeding study with Wistar rats. PMID- 15358501 TI - Determination of growth characteristics and lipolytic and proteolytic activities of Penicillium strains isolated from Argentinean salami. AB - The growth of Penicillium spp. on the surface of meat-based dry fermented sausages provides them with a protective effect against some undesirable microorganisms. Penicillium also acts as an antioxidant, minimizes the risk of excessive drying, and it is responsible for flavor development due to the decomposition of proteins, free fatty acids and lactic acid. With the aim of developing starter cultures, important physiological properties such as growth and proteolytic and lipolytic activities were evaluated on 13 mold strains belonging to the genera Penicillium. These strains were isolated from Argentinean dry fermented meat sausages named "salami". The selection was based on color, mycellium appearance and growth characteristics. The most important factors of the drying process of salami, such as temperature (14 and 25 degrees C), water activity (aw) (0.90, 0.95 and 1.00) and presence of 2.5% sodium chloride (NaCl), were analyzed. Although all strains analyzed were able to grow under the different conditions evaluated, they showed different growth velocity (K = mm/day) in response to temperature, aw and presence of NaCl in the media. All strains showed both proteolytic and lipolytic activities under the studied factors of the drying process. Nevertheless, differences in inter-species and even intra-species were found. The addition of NaCl gave a stimulant effect to the proteolytic activity at 25 degrees C, but the response at 14 degrees C was variable. The same variability was observed in the presence of salt, both at 25 and 14 degrees C, when the lipolytic activity was assayed. According to our results, detailed assays of the physiological capacities of indigenous strains proposed as starter cultures are required. PMID- 15358502 TI - Influence of water activity and temperature on growth of isolates of Aspergillus section Nigri obtained from grapes. AB - The effects of water activity (aw) and temperature on growth of Aspergillus section Nigri isolated from wine grapes were investigated on an agar medium with composition similar to that of grapes. Temperatures in the range of 10-37 degrees C were tested. Optimum temperatures for growth were between 30 and 37 degrees C. Water activity levels ranging from 0.90 to 0.995 were tested. Optimum aw for growth was 0.98 in most cases. Statistical differences were found among the groups tested (A. carbonarius, A. niger aggregate and A. section Nigri uniseriates). Growth rates models for the factors assayed have been obtained. PMID- 15358503 TI - Moulds isolated from Istrian dried ham at the pre-ripening and ripening level. AB - The aim of this study is to define the mould strains growing on the surface during the pre-ripening and the ripening phases of Istrian ham, and their toxic potential. The mould microflora was predominantly represented by five genera, which were isolated on the ham surfaces of three different producers investigated. The identified species were similar in the both tested periods, demonstrating that the contamination came mainly from the air and the ripening chambers (seasoning rooms), rather than the raw meat. Eurotium spp., Aspergillus spp. and Penicillium spp. were the main strains isolated. The presence and growth of the different strains depended on the temperature of ripening and the relative humidity in the ripening chambers, since the hams were home made products and not matured in controlled conditions. The toxic potential of isolated strains was also investigated. None of the tested moulds can produce mycotoxins and for this reason the Istrian hams do not represent a health hazard. PMID- 15358504 TI - Survival of Escherichia coli O157:H7, O111:H- and O26:H11 in artificially contaminated chocolate and confectionery products. AB - To date, the survival of Escherichia coli O157:H7 and other verocytotoxin producing E. coli (VTEC) in chocolate and other confectionery products has not been fully established, unlike Salmonella, which have been responsible for occasional outbreaks of infection linked to contaminated chocolate and related products, although none of these outbreaks have been related to products produced in the United Kingdom. The United Kingdom Biscuit, Cake, Chocolate and Confectionery Alliance commissioned this study to obtain information on the decline and potential survival of E. coli, particularly verocytotoxin-producing strains, in reduced aw confectionery products chocolate, biscuit cream and mallow. These products were artificially contaminated with high (4 log10 cfu/g) and low (2 log10 cfu/g) levels of E. coli O157:H7, O111:H- and O26:H11 and their survival, as affected by storage temperature (10, 22 and 38 degrees C), was monitored over 12 months. Preliminary studies to establish the best inoculation and recovery procedures indicated that differences between counts on selective and non-selective media used were not sufficiently different to influence the outcome of this study. Irrespective of sample type, rapid decline was observed in products stored at 38 degrees C and increased survival occurred in products stored at 10 degrees C. In chocolate (average aw 0.40), these bacteria were detected for up to 43 days in samples stored at 38 degrees C. At 22 degrees C they survived for up to 90 days and in product stored at 10 degrees C they could still be detected after 366 days storage. In biscuit cream (average aw 0.75) they survived for 2 days at 38 degrees C, 42 days at 22 degrees C and 58 days at 10 degrees C. Whilst mallow (aw ca. 0.73) was not stored at 38 degrees C, these bacteria could still be detected in samples stored for up to 113 and 273 days at 22 and 10 degrees C, respectively. The observed prolonged survival of these bacteria under conditions of reduced aw and lowered storage temperature in this study is supported by previous studies with Salmonella and E. coli O157:H7 in other foods. In the same way that Salmonella bacteria can survive for long periods, in excess of 12 months, in chocolate, this study provides evidence that E. coli, including pathogenic strains, can also survive for similar periods of time. Assuming the routes of transmission are similar, controls currently used by the confectionery industry to prevent contamination by Salmonella should also be effective against E. coli, including VT-producing strains, providing that all raw materials have been suitably processed, stored and handled before and during manufacture. PMID- 15358505 TI - Time-temperature profiles of chilled ready-to-eat foods in school catering and probabilistic analysis of Listeria monocytogenes growth. AB - The purpose of this study was to evaluate the chill chain in school catering by monitoring time-temperature profiles. Chilled ready-to-eat foods have been chosen as subject of this study because of their high risk due to their production, storage and distribution steps, separated in time, followed by consumption without any further thermal treatment. In order to integrate the effects of storage duration and storage temperature, a quantitative criterion, namely "TTE" or "Time-Temperature Equivalent", was proposed. To illustrate the sanitary consequences of the recorded thermal history, Listeria monocytogenes growth was predicted based on reference growth curves in chilled ready-to-eat food products. The study of five centralised kitchens and 11 school-lunch canteens demonstrated in general a satisfactory maintenance of the chill chain. However, the coincidence of extended storage duration (due to weekends) and temperature abuse was observed and could lead to a significant microbial development. PMID- 15358506 TI - Transition of the probiotic bacteria, Lactobacillus casei strain Shirota, in the gastrointestinal tract of a pig. AB - The transition of probiotic bacteria Lactobacillus in the gastrointestinal tract was investigated in pigs that received commercially available fermented milk prepared with Lactobacillus casei strain Shirota (LCS). Three female pigs fistulated at the cecum were fed 130 ml of fermented milk that contained over 10(10) (cfu) LCS with their daily meal for 8 days. Cecal contents were sampled through a fistula every 2 h for 24 h after marker dosing. The viable cell number (log cfu/g) of LCS and the concentrations of transit markers in each sample were determined. The viable number of LCS cells ranged from 3.56 to 6.58. The number of LCS in the cecum was not stable in pigs and varied with the flow of the cecal content. The viable number of LCS cells was significantly correlated with the relative concentration of the marker. These results indicated that most LCS moved with the liquid component. The level of LCS reached the maximum (6.38) 6 h after dosing. Four doses every 6 h may be required to maintain the maximum level of LCS at the cecum. PMID- 15358508 TI - Atmospheric oxygen and other conditions affecting the production of cereulide by Bacillus cereus in food. AB - Factors influencing the production of cereulide, the emetic toxin of Bacillus cereus in food and laboratory media were investigated, using liquid chromatography-ion trap mass spectrometry and sperm motility inhibition bioassay for detection and quantitation. Oxygen was essential for production of the emetic toxin by B. cereus. When beans, rice or tryptic soy broth were inoculated with cereulide producing strains B203, B116 (recent food isolates) or the strain F 4810/72, high amounts (2 to 7 microg ml(-1) or g(-1) wet wt) of cereulide accumulated during 4-day storage at room temperature. In parallel cultures and foods, stored under nitrogen atmosphere (> 99.5% N2), less than 0.05 microg of cereulide ml(-1) or g(-1) wet wt accumulated. The outcome of the bioassay matched that of the chemical assay, with no indication of interference by substances in the rice or beans. Boiling for 20 to 30 min did not inactivate cereulide or cereulide producing strains in rice or the beans. Adding l-leucine and l-valine (0.3 g l(-1)) stimulated cereulide production 10- to 20-fold in R2A and in rice water agar. When the B. cereus strains were grown on agar media under permissive conditions (air, room temperature), cereulide was produced overnight with little or no increase when the incubation was extended to 4 days. In broth culture, the production of cereulide started later than 16-24 h. Anoxic storage prevented cereulide production also when the amino acids had been supplied. Packaging with modified atmosphere low in oxygen may thus be used to reduce the risk of cereulide formation during storage of food. PMID- 15358507 TI - Verocytotoxin-producing Escherichia coli O157 in minced beef and dairy products in Italy. AB - A total of 3879 samples of foodstuffs were examined for the presence of Verocytotoxin-producing Escherichia coli O157 (VTEC O157). The survey was conducted by 9 of the 10 Italian Veterinary Public Health Laboratories. Samples were collected between May 2000 and September 2001 in 14 regions and comprised 931 minced beef specimens and 2948 dairy products (DP) with less than 60 days of ripening. The DP included 657 pasteurised and 811 unpasteurised bovine DP, 477 pasteurised and 502 unpasteurised ovine DP, and 501 water-buffalo's milk mozzarella cheese. Samples were collected at retail level, from plants processing minced beef and dairy plants and from farms directly manufacturing cheeses. All the samples were tested using a sensitive procedure based on ISO/DIS 16654:1999 (later ISO 16654:2001), which includes an immunomagnetic separation step. A preliminary inter-laboratory trial was organised with artificially contaminated samples to assess the ability of all the participating laboratories to isolate E. coli O157 by the established procedure. VTEC O157 was isolated from four (0.43%) of the minced beef samples, collected in four different regions and during different months, but was not detected in any of the dairy products. E. coli O157 VT-eae+ was isolated from one raw cow's milk cheese. This survey provided national data on the presence of VTEC O157 in foodstuffs, demonstrating a low prevalence of the organism. The survey also encouraged updating of knowledge and procedures on VTEC O157 in laboratories with official responsibility for microbiological testing of foods of animal origin. PMID- 15358509 TI - Molecular epidemiology and disinfectant susceptibility of Listeria monocytogenes from meat processing plants and human infections. AB - We have investigated the molecular epidemiology of Listeria monocytogenes from the meat processing industry producing cold cuts and from cases of human listeriosis by discriminative pulsed-field gel electrophoresis (PFGE). A subset of the isolates was also investigated for susceptibility to a disinfectant based on quaternary ammonium compounds (QAC) frequently used in the meat processing industry. The purpose of this investigation was to obtain knowledge of sources, routes of contamination and genetic types of L. monocytogenes present along the production line in the meat processing industry, and to compare meat industry isolates and human isolates. Of the 222 isolates from four meat-processing plants, 200 were from two plants responsible for nearly 50% of the production of cold cuts in the Norwegian market. The strain collection included historical routinely sampled isolates (1989-2002) and isolates systematically sampled through a one year period (November 2001 to November 2002) from fresh meat and production environments in three plants. No isolates were obtained in samples from employees (throat, faeces). Human strains included all available reported isolates from Norwegian patients in selected time periods. The L. monocytogenes PFGE data showed a large genetic heterogeneity, with isolates separated into two genetic lineages and further subdivided into 56 different PFGE profiles. Certain profiles were observed on both sides of production (before and after heat treatment) indicating contamination of end products by fresh meat or fresh meat environments. While fresh meat isolates almost exclusively grouped within lineage I, isolates from end products showed a more balanced distribution between lineages I and II. Ten profiles were common among isolates from human and meat industry. Typing of human isolates identified a previously unrecognised outbreak. Generally, a higher QAC resistance incidence was observed among isolates from the meat processing industry than among human isolates although large plant to plant differences were indicated. No correlation between resistance and PFGE profile or resistance and persistence was observed. PMID- 15358510 TI - The microflora of fermented nixtamalized corn. AB - Nixtamalization is a traditional process that improves the nutritional quality of corn. To provide a means of utilizing the nutritional benefits of nixtamalized corn and improve product acceptability, lactic acid fermentation was applied. The objective of the study was to study the microbial profile and establish the important lactobacilli of fermenting nixtamalized corn dough. Two batches of cleaned whole corn were subjected to the process of nixtamalization, using two concentrations of lime (0.5 or 1.0%), milled, made into a dough (50% moisture) and fermented spontaneously for 72 h. A control sample was prepared without alkaline treatment. pH and titratable acidity of the dough were measured. Aerobic mesophiles, lactic acid bacteria, yeasts and molds were enumerated on Plate Count Agar (PCA), deMan, Rogossa and Sharpe (MRS) Agar and Malt Extract Agar (MEA), respectively. The identity of lactobacilli present was established at the species level using API 50 CHL. The pH of all the fermenting systems decreased with fermentation time with concomitant increase in titratable acidity. Lactic acid bacteria in numbers of 1.6 x 10(9), 2.3 x 10(9) and 1.8 x 10(9) cfu/g, respectively yeasts and molds, and numbers of 8.0 x 10(7), 5.0 x 10(5) and 1.7 x 10(5) cfu/g, respectively were observed in the control and the two nixtamalized (0.5% and 1.0% lime) samples after 48 h of fermentation. Lactobacilli identified in the fermenting nixtamalized corn dough were Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus cellobiosus as well as Pediococcus spp. The study demonstrates that nixtamalized corn though alkaline in nature can be subjected to spontaneous fermentation to produce a sour product. PMID- 15358511 TI - Attachment of Escherichia coli O157:H7 grown in tryptic soy broth and nutrient broth to apple and lettuce surfaces as related to cell hydrophobicity, surface charge, and capsule production. AB - This study investigated the effect of growth in tryptic soy broth (TSB) and nutrient broth (NB) on the ability Escherichia coli O157:H7 to attach to lettuce and apple surfaces. In addition, cell surface hydrophobicity, charge and capsule production were determined on cells grown in these media. Cells grown in NB attached less to lettuce and apple surfaces than did those grown in TSB. TSB, but not NB, supported capsule production by E. coli O157:H7. Cells grown in TSB were more hydrophilic than those grown in NB. No difference was found in the electrokinetic properties of cells grown in these media. Electrostatic and hydrophobic interactions and surface proteins did not appear to play an important role in the attachment of E. coli O157:H7 to these surfaces. Of the factors studied, only capsule production was associated with attachment ability. PMID- 15358512 TI - Chromatin structure at the flanking regions of the human beta-globin locus control region DNase I hypersensitive site-2: proposed nucleosome positioning by DNA-binding proteins including GATA-1. AB - The human beta-globin locus control region DNase I hypersensitive site-2 (LCR HS 2) is erythroid-specific and is located 10.9 kb upstream of the epsilon-globin gene. Most studies have only examined the core region of HS-2. However, previous studies in this laboratory indicate that positioned nucleosomes are present at the 5'- and 3'-flanking regions of HS-2. In addition, footprints were observed that indicated the involvement of DNA-binding proteins in positioning the nucleosome cores. A consensus GATA-1 site exists in the region of the 3' footprint. In this study, using an electrophoretic mobility shift assay (EMSA) and DNase I footprinting, we confirmed that GATA-1 binds in vitro at the 3'-end of HS-2. An additional GATA-1 site was found to bind GATA-1 in vitro at a site positioned 40 bp upstream. At the 5'-end of HS-2, DNase I footprinting revealed a series of footprints showing a marked correlation with the in vivo footprints. EMSA indicated the presence of several erythroid-specific complexes in this region including GATA-1 binding. Sequence alignment for 12 mammalian species in HS-2 confirmed that the highest conservation to be in the HS-2 core. However, a second level of conservation extends from the core to the sites of the proposed positioning proteins at the HS-2 flanking regions, before declining rapidly. This indicates the importance of the HS-2 flanking regions and supports the proposal of nucleosome positioning proteins in these regions. PMID- 15358513 TI - Characterization of GhRac1 GTPase expressed in developing cotton (Gossypium hirsutum L.) fibers. AB - Cytoskeleton assembly plays an important role in determining cotton fiber cell length and morphology and is developmentally regulated. As in other plant cells, it is not clear how cytoskeletal assembly in fibers is regulated. Recently, several Rac/Rop GTPases in Arabidopsis were shown to regulate isotropic and polar cell growth of root hairs and pollen tubes by controlling assembly of the cytoskeleton. GhRac1, isolated from cottonseeds, is a member of the Rac/Rop GTPase family and is abundantly expressed in rapidly growing cotton tissues. GhRac1 shows the greatest sequence similarity to the group IV subfamily of Arabidopsis Rac/Rop genes. Overexpression of GhRac1 in E. coli led to the production of a functional GTPase as shown by in vitro enzyme activity assay. In contrast to other Rac/Rop GTPases found in cotton fiber, GhRac1 is highly expressed during the elongation stage of fiber development with expression decreasing dramatically when the rate of fiber elongation declines. The association of highest GhRac1 expression during stages of maximal cotton fiber elongation suggests that GhRac1 GTPase may be a potential regulator of fiber elongation by controlling cytoskeletal assembly. PMID- 15358514 TI - Protease-sensitive signalling by chemically engineered intramolecular fluorescent resonance energy transfer mutants of green fluorescent protein. AB - The native cysteine residues of green fluorescent protein (GFP) at positions 48 and 70 were replaced by non-thiolic amino acids, and new cysteine sites were introduced at specific, surface positions. Based on molecular modeling of the GFP structure, the sites chosen for mutagenesis to Cys were glutamic acid at position 6 and isoleucine at position 229. These new, unique cysteine sites provided reactive thiol groups suitable for site-specific chemical modification by eosin based fluorescence labels. The new constructs were designed to serve as the basis of proof of principle for fluorescence resonance energy transfer (FRET) using an enzyme-activated (trypsin) intervening sequence between native and chemically conjugated fluorophores. These eosin moieties provided chemical FRET partners for the native GFP chromophore. On excitation, these GFP-eosin constructs exhibited strong intramolecular FRET, with quenching of the native GFP (511 nm) fluorophore emission and emission around 540 nm, corresponding to eosin. GFP mutants engineered with trypsin-sensitive sequences close to the eosin site, so that on trypsinolysis FRET was destroyed, the emission wavelength switching from that of the chemical FRET partner back to that of the native GFP fluorophore, providing efficient, ratio-based detection. This protein engineering provides the basis for novel bioprobes for enzymatic triggering using intramolecular FRET between GFP and carefully sited chemical labels. PMID- 15358515 TI - Structure of primate and rodent orthologs of the prostate cancer susceptibility gene ELAC2. AB - The human ELAC2 gene was the first candidate prostate cancer susceptibility gene identified by linkage analysis and positional cloning. DNA sequence indicates a protein of 826 amino acids encoded by 24 exons. In the present study, we characterized the coding sequence of chimpanzee and gorilla ELAC2 orthologs by direct sequencing of genomic fragments, and of cynomolgus monkey and rat orthologs by screening cDNA libraries. The orthologs characterized in the chimpanzee, gorilla and cynomolgus monkey also encode proteins of 826 amino acids, sharing 98.9%, 98.5% and 93.7% sequence identity with the human protein. Our analyses of the mouse ELAC2 gene identified two alternative mRNA transcripts. One is translated into a protein of 824 a.a. (mouse ELAC2), whereas the other one encodes a protein of 831 amino acids (mouse ELAC2A) resulting from an alternatively spliced form of 25 exons. The rat ELAC2 gene ortholog also expressed two similar alternatively spliced transcripts. These two forms are ubiquitously expressed in mouse and rat tissues. The highest levels of expression of the ELAC2 form are observed in the testis while the lowest levels are seen in the prostate and in the muscle. However, it is of interest to note that the relative abundance of the rat and mouse ELAC2 transcripts, measured by real-time quantitative PCR, is higher than the respective ELAC2A forms in all surveyed tissues except for the prostate and the muscle. The ELAC2A transcript levels are 4.1 to 5.0-fold higher than the ELAC2 levels in the prostate of rat and mouse, respectively. A fine analysis of the conserved domains on the primary structure of ELAC2 orthologs revealed the presence of a putative beta-CASP domain shared by the PSO2 (SNM1) DNA interstrand cross-link repair proteins, and the 73-kDa subunit of mRNA 3' end cleavage and polyadenylation specificity factor (CPSF73) as well as Artemis proteins, thus suggesting a potential interaction of ELAC2 gene product with nucleic acids and more specifically with RNA targets. Taken together, these data offer useful tools to further study the regulation and cellular function of ELAC2 gene in experimental models and provide further insight concerning conserved amino acid motifs that could have biological significance. PMID- 15358516 TI - Regulation of Gene33 expression by insulin requires MEK-ERK activation. AB - Gene33 and its human homologue, mitogen inducible gene-6/receptor-associated late transducer (mig-6, RALT), is a 53-kDa soluble protein that was identified as a hepatic gene regulated by glucocorticoids and insulin. Its mRNA is expressed in numerous tissues in addition to the liver. Mitogen inducibility of Gene33 mRNA has been described in several experimental systems. Recent reports have suggested a role for Gene33 in inhibition of proliferation induced by factors that bind to members of the ErbB family of receptors. In the present work, we examine the regulation of Gene33 protein by insulin in hepatoma cells of rat (H4IIE) and human (HepG2/Hep3B) origin. Inhibition of MEK1 significantly inhibited extracellularly regulated kinase (ERK)1/2 activation and insulin-regulated Gene33 transcription and protein levels in H4IIE cells. Inhibition of phosphatidylinositol 3-kinase (PI3-K) activity alone did not significantly alter transcription of Gene33. In Hep3B and HepG2 cells, insulin did not significantly induce either ERK1/2 activation or Gene33 expression. This work suggests that the MEK-ERK, but not the phosphatidylinositol 3-kinase (PI3-K), pathway plays a direct role in insulin regulation of Gene33 transcription and protein expression. PMID- 15358517 TI - Armadillo/Pangolin regulates PCNA and DREF promoter activities. AB - Here we show that Armadillo and Pangolin (dTCF), downstream effectors of the Wingless (Wg) signal transduction pathway, activate transcription of the important DNA replication-related genes encoding Drosophila proliferating cell nuclear antigen (PCNA) and DNA replication-related element-binding factor (DREF). By transient luciferase expression assays and band mobility shift assays, we demonstrated the PCNA gene to be a direct target gene for the Armadillo/Pangolin complex. Using a GAL4-UAS system, stimulation of the PCNA gene by Armadillo/Pangolin was confirmed in adult females. From the published reports of an inhibitory role, we expected that Drosophila CREB-binding protein (dCBP) would interfere with activation. However, effects were only observed with the DREF but not the PCNA gene. In the latter case, as in mammals, dCBP could potentiate Armadillo-mediated activation. These results suggest that first, PCNA and DREF genes are targets of the Armadillo/Pangolin complex and second, dCBP modulates Wg signaling in a gene-specific manner. PMID- 15358518 TI - Effects of dexamethasone, vitamin A and vitamin D3 on DSP-PP mRNA expression in rat tooth organ culture. AB - Vitamin A, 1,25-dihydroxyvitamin D3 and dexamethasone are well-characterized hydrophobic molecules whose biological actions are mediated via different members of the nuclear hormone receptor family. We report here their actions on tooth formation at the molecular level. We have tested the effects of these compounds on osteopontin (OPN), dentin sialoprotein (DSP-PP), and collagen type I expression in pre-mineralization and mineralization stage rat tooth organ cultures which mirror in vivo developmental patterns. These proteins are all believed to participate in the mineralization of dentin. 1,25-Dihydroxyvitamin D3 up-regulated OPN, but had no effect on DSP-PP mRNA expression. Vitamin A up regulated DSP-PP expression as did dexamethasone. Dexamethasone also up-regulated collagen type I expression. Our results suggest that 1,25-dihydroxyvitamin D3 does not modulate dentin mineralization by directly affecting DSP-PP expression. Vitamin A likely contributes to dentin mineralization by up-regulating DSP-PP expression. Finally, the up-regulation of DSP-PP expression in tooth germ cultures treated with dexamethasone suggests that its application to patient's dental pulp might promote increased extracellular matrix synthesis and mineralization in the pulp and may explain the narrowing of the dental pulp cavity in patients undergoing long-term dexamethasone administration. PMID- 15358519 TI - Identification of the functional elements in the promoter region of human DNA topoisomerase IIIbeta gene. AB - In this study, we have isolated and characterized the promoter region of the human DNA topoisomerase IIIbeta (hTOP3beta) gene. The 5' RACE assay showed a short exon 1 encoding only the 35-bp untranslated region and suggested the presence of multiple transcription initiation sites. The hTOP3beta gene promoter lacks a canonical TATA box or initiation element and is moderately high in GC content. Transient expression of a luciferase reporter gene under the control of serially deleted 5'-flanking sequence identified an activator element between 141 and -119 upstream of the transcription initiation site and a second regulatory element between -91 and -71. On the basis of scanning mutations of triple nucleotides, we demonstrated that a 5'GGAACC3' element between -117 and 112 plays a critical role in the up-regulation of the basal transcription activity. Changing the 5'GGAACC3' sequence leads to markedly reduced promoter activity. Gel mobility shift assays revealed that the 5'GGAACC3' element is required for DNA binding by the transcription factor complex. These observations lead to the conclusion that the positive regulatory region including the 5'GGAACC3' core element is essential for efficient expression of the hTOP3beta gene as well as for the binding of as yet unidentified regulatory factor(s). PMID- 15358520 TI - Elicitor-induced activation of transcription via W box-related cis-acting elements from a basic chitinase gene by WRKY transcription factors in tobacco. AB - A putative elicitor responsive element with two W boxes (CTGACC/T) has been identified in the region between -125 and -69 of a tobacco class I basic chitinase gene CHN48. We generated transgenic tobacco calli that contained the 125/-69 region fused to a luciferase reporter gene. The expression of the reporter gene was induced upon treatment with an elicitor, xylanase from Trichoderma viride (TvX). This induction required protein kinase activity. We isolated three cDNA clones encoding DNA-binding proteins, designated as NtWRKY1, NtWRKY2, and NtWRKY4, from tobacco cultured cells. Gel mobility shift assays showed that in vitro translation products of NtWRKY1, NtWRKY2 and NtWRKY4 bound to W box of CHN48 gene. These NtWRKY proteins stimulated W box-mediated transcription of a luciferase reporter gene in the transient assay. In addition, the transactivation of W box-mediated transcription by NtWRKY1 and NtWRKY4 was enhanced in response to elicitor treatment, suggesting elicitor-induced posttranscriptional activation of these NtWRKYs. Northern blot analyses showed that mRNAs for NtWRKY1 and NtWRKY2 increased after treatment with the elicitor, whereas mRNAs for NtWRKY4 were expressed constitutively at a low level. These results suggested possible involvement of NtWRKYs in elicitor-responsive transcription of defense genes in tobacco. PMID- 15358521 TI - Identification of the critical cis-acting elements in the promoter of the mouse Prnd gene coding for Doppel protein. AB - Prnd gene encodes a prion-like protein Doppel. To elucidate the transcriptional regulation of the mouse Prnd gene, we performed a study of its 5' flanking region. Serial deletion analysis of the -1863/+27 genomic fragment revealed the region -185/+27 as a core promoter. Cis-acting elements at the -182/-177 (E box) and -108/-104 (CCAAT box) positions were identified as the main activators of the Prnd expression in GC1-spg cells, with NF-Y being the important contributor. The gel-shift experiments confirmed the binding of the transcription factors to -108/ 104 NF-Y consensus motif. An intron1 was found to enhance the Prnd expression in bEND.3 endothelial cells, suggesting an involvement of the endothelial-specific enhancer. PMID- 15358522 TI - Space distribution of harmonic mode vibration amplitudes in nonlinear finite piezoelectric transducer. AB - Nonlinear elastic vibrations of cylindrical piezoelectric transducers are investigated both experimentally and theoretically. A particular behaviour, that relates the space distribution of the fundamental mode vibration to those of the second and third harmonic components, is observed. A simplified physical interpretation of the phenomenon is given. PMID- 15358523 TI - Analysis of ultrasonic attenuation in particle-reinforced plastics by a differential scheme. AB - Attenuation of ultrasonic longitudinal waves in some particle-reinforced polymer composites is studied theoretically by a micromechanical model based on a differential (incremental) scheme. A set of differential equations is established by which the attenuation spectrum of the composite can be computed from the known properties of viscoelastic matrix and elastic particles. For a composite reinforced with glass particles with radius 0.15 mm, the proposed scheme is shown to predict the attenuation in better agreement with the foregoing experimental results than the previous simplistic independent scattering model. Based on this scheme, the dependence of the longitudinal attenuation spectrum of a particulate polymer composite on the wavelength-to-particle radius ratio and the particle volume fraction is examined in detail. It is then shown theoretically that the attenuation of the composite decreases monotonically with the particle volume fraction when the particle radius is sufficiently small compared to the incident wavelength, while it shows non-monotonic particle-fraction dependence when the ratio of the particle radius to the wavelength is larger. To examine this theoretical finding from an experimental point of view, the longitudinal attenuation in a glass-particle-reinforced polyester composite with particle radius 0.0225 mm is measured for different particle volume fractions. The measured attenuation characteristics are shown to support the qualitative features of the theoretical prediction. PMID- 15358524 TI - Identification of the piezoelectric material coefficients using the finite element method with an asymptotic waveform evaluation. AB - A rapid identification of the piezoelectric material constants for a piezoelectric transducer is proposed. The validity of a three-dimensional finite element routine was confirmed experimentally. The asymptotic waveform evaluation (AWE) was adopted for a fast frequency sweep of the finite element analysis. The three-dimensional finite element method with an AWE and a design sensitivity method was used for a material inversion scheme of piezoelectric transducers. In order to confirm the inversion routine of the material constants, the mechanical displacements, which mean the mode shape, were calculated along the vertical and lateral position of the sample transducer. PMID- 15358525 TI - Numerical simulation of particle motion in an ultrasound field using the lattice Boltzmann model. AB - In this paper we investigate the motion of small particles suspended in a fluid through which an ultrasound field is propagating. The application of the lattice Boltzmann model to this problem is considered using a two dimensional model. Particles in an ultrasound field are observed to move with a mean particle motion. Further, the time-averaged force on a fixed cylinder is computed and found to be in good agreement with a theoretical expression for the radiation force. Simulations are performed with a single particle, although the approach can equally be applied for a larger number of particles. PMID- 15358526 TI - On the splitting of aspherical structure submitted to an acoustic polarized excitation. AB - We experimentally show how the deviations from spherical symmetry such as the inhomogeneity, the removal mass or the asphericity affect the frequency spectrum of a single elastic sphere. The recorded spectrograms point out the great influence of the polarization of the source on the splitting and the shift of the peaks. Qualitatively, the results are consistent with the data reported in Geophysics although the present study is concerned with a low degree and low- [Formula: see text] modes in contrast to those usually encountered in seismic investigations. From a practical point of view, this study suggests that Geophysics phenomena could be analyzed through spherical beads reproducing the imperfections encountered in the mantle and in the core of the earth. PMID- 15358527 TI - Design and construction of shaft-driving type piezoceramic ultrasonic motor. AB - A new approach in design of shaft-driving type piezoceramic ultrasonic motor is proposed. The stator of motor consisted of a commercial available buzzer disk in which a piezoceramic membrane is adhered to a metal sheet. The wave propagation on the metal sheet was generated by extended-shrunk force from piezoceramic oscillation. Driving energy came from the vibration modes by mechanical electrical oscillation of the metal sheet in corresponding to converse piezoelectric effect using a single-phase AC voltage power. Where the relative elliptic motion was occurred between the bearing seat and rotor in order to kinematical delivery, the rotor being driven was connected directly on the bearing seat to transmit the dynamic power with frictional contact force. In analysis of dynamic features, the system transfer function of admittance and equivalent circuit was obtained. The rotating speed of the prototype motor could be reached as high as 2000 rpm on the driving condition of 72 kHz, +/-10 V(pp), and 0.2 A. The maximum torque was less than 0.003 Nm. It could be utilized in the driver of CD, or the cooling fan in the computer CPU. PMID- 15358528 TI - A novel ultrasonic clutch using near-field acoustic levitation. AB - This paper investigates design, fabrication and drive of an ultrasonic clutch with two transducers. For the two transducers, one serving as a driving element of the clutch is connected to a driving shaft via a coupling, and the other serving as a slave element of the clutch is connected to a slave shaft via another coupling. The principle of ultrasonic levitation is first expressed. Then, a series-resonant inverter is used to generate AC voltages at input terminals of each transducer, and a speed measuring system with optic sensors is used to find the relationship between rotational speed of the slave shaft and applied voltage of each transducer. Moreover, contact surfaces of the two transducers are coupled by the frictional force when both the two transducers are not energized, and separated using the ultrasonic levitation when at least one of the two transducers is energized at high voltages at resonance. PMID- 15358529 TI - Improving motion estimation by accounting for local image distortion. AB - Cardiac elastography is a useful diagnostic technique for detection of heart function abnormalities, based on analysis of echocardiograms. The analysis of the regional heart motion allows assessing the extent of myocardial ischemia and infarction. In this paper, a new two-stage algorithm for cardiac motion estimation is proposed, where the data is taken from a sequence of 2D echocardiograms. The method combines the advantages of block-matching and optical flow techniques. The first stage employs a standard block-matching algorithm (sum of absolute differences) to provide a displacement estimate with accuracy of up to one pixel. At the second stage, this estimate is corrected by estimating the parameters of a local image transform within a test window. The parameters of the image transform are estimated in the least-square sense. In order to account for typical heart motions, like contraction/expansion, translation and rotation, a local affine model is assumed within the test window. The accuracy of the new algorithm is evaluated using a sequence of 500 grayscale B-mode images, which are generated as distorted, but known copies of an original ROI, taken from a real echocardiogram. The accuracy of the motion estimation is expressed in terms of errors: maximum absolute error, root-mean-square error, average error and standard deviation. The errors of the proposed algorithm are compared with these of the known block-matching technique with cross-correlation and interpolation in the sub-pixel space. Statistical analysis of the errors shows that the proposed algorithm provides more accurate estimates of the heart motion than the cross correlation technique with interpolation in the sub-pixel space. PMID- 15358530 TI - Nuclear phosphoproteins HMGA and their relationship with chromatin structure and cancer. AB - The structural characteristics of the three nuclear phosphoproteins of the high mobility group A family are outlined and related to their participation in chromatin structure alteration in many biological processes such as gene expression, neoplastic transformation, differentiation, and apoptosis. The elevated expression of these proteins in tumor cells and their post-translational modifications, such as phosphorylation, acetylation and methylation, are discussed and suggested as suitable targets for cancer chemotherapy. PMID- 15358531 TI - Coupling between vesicle shape and the non-homogeneous lateral distribution of membrane constituents in Golgi bodies. AB - In this work, a hypothesis is presented that could explain the non-homogeneous lateral distribution of membrane components in Golgi vesicles. It is shown that the non-homogeneous lateral distribution of membrane components and the specific flattened shape of Golgi vesicles are strongly coupled. In agreement with experimental evidence, it is indicated that some of the membrane components may be concentrated mainly on the curved bulbous rims of the Golgi vesicles, while the other components are distributed predominantly in their flat central part. PMID- 15358532 TI - The death domain protein p84N5, but not the short isoform p84N5s, is cell cycle regulated and shuttles between the nucleus and the cytoplasm. AB - P84N5 is a death domain containing protein that interacts with the tumor suppressor retinoblastoma protein and induces apoptosis. We cloned and characterized two novel alternatively spliced versions of p84N5. The p84N5 short isoform (p84N5s) lacks the death domain and does not induce apoptosis. We showed that p84N5, but not p84N5s, is cell cycle regulated. We found that p84N5-GFP chimera can rapidly shuttle between the nucleus and the cytoplasm. Taken together, these observations suggest that p84N5 may transmit signals from the nucleus to cytoplasmic effectors. PMID- 15358533 TI - Auto-inhibition of Arabidopsis thaliana plasma membrane Ca2+-ATPase involves an interaction of the N-terminus with the small cytoplasmic loop. AB - Type IIB Ca2+-ATPases have a terminal auto-inhibitory, domain the action of which is suppressed by calmodulin (CaM) binding. Here, we show that a peptide (6His-1M I116) corresponding to the first 116 aminoacids (aa) of At-ACA8, the first cloned isoform of Arabidopsis thaliana plasma membrane Ca2+-ATPase, inhibits the activity of the enzyme deprived of the N-terminus by controlled trypsin treatment 10-fold more efficiently than a peptide (41I-T63) corresponding only to the CaM binding site. A peptide (268E-W348) corresponding to 81 aa of the small cytoplasmic loop of At-ACA8 binds peptide 6His-1M-I116 immobilized on Ni-NTA agarose. Peptide 268E-W348 stimulates Ca2+-ATPase activity. Its effect is not additive with that of CaM and is suppressed by tryptic cleavage of the N terminus. These results provide the first functional identification of a site of intramolecular interaction with the terminal auto-inhibitory domain of type IIB Ca2+-ATPases. PMID- 15358534 TI - Oligomerization activity of a double-stranded RNA-binding domain. AB - Xenopus laevis RNA-binding protein A (Xlrbpa) is a highly conserved, ubiquitously expressed hnRNP- and ribosome-associated RNA-binding protein that contains three double stranded RNA-binding domains (dsRBDs) in tandem arrangement. A two-hybrid screen with Xlrbpa as a bait recovered Xlrbpa itself as the strongest interaction partner, indicating multimerization of this protein. To search for regions responsible for the observed interaction, we conducted two-hybrid assays with Xlrbpa deletion constructs and identified the third dsRBD of Xlrbpa as the exclusive interacting domain. Additionally, these results were confirmed by coimmunoprecipitation experiments with truncated proteins expressed both in yeast and Xenopus oocytes. In PACT, the human homologue of Xlrbpa, we could demonstrate that the third dsRBD displays the same multimerization activity. Interestingly, this domain is essential for the activation of the dsRNA-activated protein kinase PKR. Addition of RNAses to coimmunoprecipitation experiments did not affect the dimerization, suggesting that the interaction is independent of RNA-binding. We report here a homomultimerization activity of a type B dsRBD and suggest possible implications that include a model for PKR activation by PACT. PMID- 15358535 TI - Aquaporin homologues in plants and mammals transport ammonia. AB - Using functional complementation and a yeast mutant deficient in ammonium (NH4+) transport (Deltamep1-3), three wheat (Triticum aestivum) TIP2 aquaporin homologues were isolated that restored the ability of the mutant to grow when 2 mM NH4+ was supplied as the sole nitrogen source. When expressed in Xenopus oocytes, TaTIP2;1 increased the uptake of NH4+ analogues methylammonium and formamide. Furthermore, expression of TaTIP2;1 increased acidification of the oocyte-bathing medium containing NH4+ in accordance with NH3 diffusion through the aquaporin. Homology modeling of TaTIP2;1 in combination with site directed mutagenesis suggested a new subgroup of NH3-transporting aquaporins here called aquaammoniaporins. Mammalian AQP8 sharing the aquaammoniaporin signature also complemented NH4+ transport deficiency in yeast. PMID- 15358536 TI - PD-1 inhibits T-cell receptor induced phosphorylation of the ZAP70/CD3zeta signalosome and downstream signaling to PKCtheta. AB - Engagement of the immunoinhibitory receptor, programmed death-1 (PD-1) attenuates T-cell receptor (TCR)-mediated activation of IL-2 production and T-cell proliferation. Here, we demonstrate that PD-1 modulation of T-cell function involves inhibition of TCR-mediated phosphorylation of ZAP70 and association with CD3zeta. In addition, PD-1 signaling attenuates PKCtheta activation loop phosphorylation in a cognate TCR signal. PKCtheta has been shown to be required for T-cell IL-2 production. A phosphorylated PD-1 peptide, corresponding to the C terminal immunoreceptor tyrosine-switch motif (ITSM), acts as a docking site in vitro for both SHP-2 and SHP-1, while the phosphorylated peptide containing the N terminal PD-1 immunoreceptor tyrosine based inhibitory motif (ITIM) associates only with SHP-2. PMID- 15358537 TI - Involvement of MPK4 in osmotic stress response pathways in cell suspensions and plantlets of Arabidopsis thaliana: activation by hypoosmolarity and negative role in hyperosmolarity tolerance. AB - Three of the protein kinases activated by hypoosmotic stress in Arabidopsis thaliana cell suspensions were previously characterized [FEBS, 2002, 527, 43-50] as mitogen-activated protein (MAP) kinases and two of them corresponded to Arabidopsis mitogen-activated protein kinase 6 (MPK6) (44 kDa) and MPK3 (39 kDa). The third MAP kinase was identified here to MPK4, using a corresponding specific antibody. Like MPK6 and MPK3, MPK4 activity is clearly inhibited by apigenin and MPK4 activation by hypoosmolarity needs upstream phosphorylation events. Activation of the 3 MAP kinases, MPK3, 4 and 6, was confirmed in plantlets submitted to hypoosmotic stress. The action of a biotic signal, flagellin, was also demonstrated to induce the activations of the 3 MAP kinases. Using the mutant displaying MPK4 gene inactivation, the independence of the MPK3 and MPK6 activations towards the presence of MPK4 was demonstrated, both in hypoosmotic and flagellin signalling pathways. Although MPK4 was not activated by hyperosmolarity in cell suspensions nor in seedlings, a possible negative regulation of hyperosmolarity resistance by MPK4 is suggested, based both on phenotype and downstream gene expression studies. PMID- 15358538 TI - Crystallographic studies of shikimate binding and induced conformational changes in Mycobacterium tuberculosis shikimate kinase. AB - The X-ray crystal structure of Mycobacterium tuberculosis shikimate kinase (SK) with bound shikimate and adenosine diphosphate (ADP) has been determined to a resolution of 2.15 A. The binding of shikimate in a shikimate kinase crystal structure has not previously been reported. The substrate binds in a pocket lined with hydrophobic residues and interacts with several highly conserved charged residues including Asp34, Arg58, Glu61 and Arg136 which project into the cavity. Comparisons of our ternary SK-ADP-shikimate complex with an earlier binary SK-ADP complex show that conformational changes occur on shikimate binding with the substrate-binding domain rotating by 10 degrees. Detailed knowledge of shikimate binding is an important step in the design of inhibitors of SK, which have potential as novel anti-tuberculosis agents. PMID- 15358539 TI - Transport of somatostatin and substance P by human P-glycoprotein. AB - P-glycoprotein is an efflux pump for a broad spectrum of hydrophobic agents. We found that bioactive peptides including somatostatin and substance P inhibit ATP dependent vincristine binding to P-glycoprotein-overexpressing K562/ADM membrane vesicles. Some of these bioactive peptides including somatostatin stimulate basal ATPase activity of P-glycoprotein; in contrast, other peptides including substance P inhibit it. The K562/ADM membrane vesicles showed an ATP-dependent, osmotically sensitive uptake of somatostatin and substance P, which was inhibited by valspodar, an inhibitor of P-glycoprotein. These findings suggested that certain bioactive peptides such as somatostatin and substance P directly interact with human P-glycoprotein as endogenous substrates for P-glycoprotein-mediated transport. PMID- 15358540 TI - Functional identification of AtFao3, a membrane bound long chain alcohol oxidase in Arabidopsis thaliana. AB - The Arabidopsis thalina genome database was searched for homologues of the Candida cloacae fao1 gene which encodes a membrane bound, flavin-containing, hydrogen peroxide generating, long chain alcohol oxidase. This gene has not been isolated from plants or animals. Four putative candidates were found in the database but their function has not been proven. The cDNAs for two of them were cloned by RT-PCR from Arabidopis suspension culture and one of them [AtFAO3] was overexpressed in Escherichia coli and shown to functionally express long chain alcohol oxidase activity. The protein has been solubilised and retains biological activity thereby preparing the way for crystallographic studies. This is the first functional proof identifying a long chain alcohol oxidase in higher plants. PMID- 15358541 TI - The role of the cyclic peptide backbone in the anti-HIV activity of the cyclotide kalata B1. AB - The plant cyclotides, the largest known family of circular proteins, have tightly folded structures and a range of biological activities that lend themselves to potential pharmaceutical and agricultural applications. Based on sequence homology, they are classified into the bracelet and Mobius subfamilies. The bracelet subfamily has previously been shown to display anti-HIV activity. We show here that a member of the Mobius subfamily, kalata B1, also exhibits anti HIV activity despite extensive sequence differences between the subfamilies. In addition, acyclic permutants of kalata B1 displayed no anti-HIV activity, suggesting that this activity is critically dependent on an intact circular backbone. PMID- 15358542 TI - Inhibition of membrane-bound lytic transglycosylase B by NAG-thiazoline. AB - The lytic transglycosylases cleave the bacterial cell wall heteropolymer peptidoglycan with the same specificity as the muramidases (lysozymes), between the N-acetylmuramic acid and N-acetylglucosamine residues, with the concomitant formation of a 1,6-anhydromuramoyl residue. The putative catalytic residue in the family 3 lytic transglycosylase from Pseudomonas aeruginosa, Glu162 as identified by sequence alignment to the homologous enzyme from Escherichia coli, was replaced with both Ala and Asp by site-directed mutagenesis. Neither mutant enzyme differed structurally from the wild-type enzyme, as judged by CD spectroscopy, but both were enzymatically inactive confirming the essential role of Glu162 in the mechanism of action of this lytic transglycosylase. The beta hexosaminidase inhibitor NAG-thiazoline was shown to inhibit the activity of lytic transglycosylase activity, thus providing the first direct evidence that the formation of the 1,6-anhydromuramoyl residue may proceed through an oxazolinium ion intermediate involving anchimeric assistance. Using surface plasmon resonance and difference absorbance spectroscopy, Kd values of 1.8 and 1.4 mM, respectively, were determined for NAG thiazoline, while its parent compound N-acetylglucosamine neither inhibited nor appeared to bind the lytic transglycosylase with any significant affinity. PMID- 15358543 TI - Gene delivery of the elastase inhibitor elafin protects macrophages from neutrophil elastase-mediated impairment of apoptotic cell recognition. AB - The resolution of inflammation is dependent on recognition and phagocytic removal of apoptotic cells by macrophages. Receptors for apoptotic cells are sensitive to degradation by human neutrophil elastase (HNE). We show in the present study that HNE cleaves macrophage cell surface CD14 and in so doing, reduces phagocytic recognition of apoptotic lymphocytic cells (Mutu 1). Using an improved method of adenovirus-mediated transfection of macrophages with the HNE inhibitor elafin, we demonstrate that elafin overexpression prevents CD14 cleavage and restores apoptotic cell recognition by macrophages. This approach of genetic modification of macrophages could be used to restore apoptotic cell recognition in inflammatory conditions. PMID- 15358544 TI - Carotenoid-induced quenching of the phycobilisome fluorescence in photosystem II deficient mutant of Synechocystis sp. AB - Brief--10-second long--irradiation of a photosystem II-deficient mutant of cyanobacterium Synechocystis sp. PCC 6803 with intense blue or UV-B light causes an about 40% decrease of phycobilisome (PBS) fluorescence, slowly reversible in the dark. The registered action spectrum of PBS fluorescence quenching only shows bands at 500, 470 and 430 nm, typical of carotenoids, and an additional UV-B band; no peaks in the region of chlorophyll or PBS absorption have been found. We propose that quenching induced by carotenoids, possibly protein-bound or glycoside, reveals a new regulatory mechanism protecting photosynthetic apparatus of cyanobacteria against photodamage. PMID- 15358545 TI - Plasmin cleaves the juxtamembrane domain and releases truncated species of the urokinase receptor (CD87) from human bronchial epithelial cells. AB - The three-domain (D1D2D3) urokinase receptor (CD87) is highly susceptible to cleavage within the D1-D2 linker sequence, but also within the juxtamembrane region by yet poorly characterized proteinases, allowing the release of D1 and D2D3 species in various (patho)physiological body fluids. Using immunoblot analysis and ELISA applied to a recombinant soluble CD87 and to CD87-expressing epithelial cells, we establish that exogenous or in situ generated plasmin proteolyzes CD87 in the D1-D2 linker and D3 carboxyterminal sequences, producing a major soluble D2D3 species. Mass spectrometry analysis of the fragmentation of CD87-related synthetic peptides, and aminoterminal sequencing of D2D3 reveal Arg83, Arg89, and Arg281 as residues targeted by plasmin within human CD87. PMID- 15358546 TI - Binding of von Willebrand factor to the small proteoglycan decorin. AB - The small proteoglycan decorin plays an important role in the organisation of the extracellular matrix by binding to several components, including collagen and fibronectin. In this work, we report the dose-dependent and saturable interaction of decorin with the adhesive glycoprotein, von Willebrand factor (VWF). This interaction was mediated by the glycosaminoglycan side chain of decorin and was critically regulated by the degree of sulfation, but not by the amount of iduronic acid. Both chondroitin sulfate and dermatan sulfate, in addition to heparin, were found to bind VWF equally well. Although soluble decorin prevented VWF binding to heparin, purified VWF-A1 domain failed to interact with the proteoglycan. These results identify VWF as a new partner for the small proteoglycan, decorin, in the structural organisation of the extracellular matrix. PMID- 15358547 TI - A cyanobacterial gene encoding an ortholog of Pirin is induced under stress conditions. AB - Pirin is a recently identified protein in eukaryotes as a transcription cofactor or as an apoptosis-related protein. Although Pirin is highly conserved from bacteria to human, there have been no reports on prokaryotic Pirin orthologs. We show here that pirA (sll1773) encoding an ortholog of Pirin together with an adjacent gene, pirB (ssl3389), was upregulated under high salinity and some other stress conditions in a cyanobacterium Synechocystis sp. PCC 6803. Induction of the pirAB genes was not related to cell death and disruption of pirA did not affect the gene expression profile. Expression of the pirAB genes was negatively regulated by a LysR family transcriptional regulator encoded by pirR (slr1871) located immediately upstream of pirAB in the divergent direction. DNA microarray analysis indicated that PirR repressed expression of closely located ORFs, slr1870 and mutS (sll1772), in addition to pirAB and pirR itself. PMID- 15358548 TI - Angiotensin II-induced apoptosis in human endothelial cells is inhibited by adiponectin through restoration of the association between endothelial nitric oxide synthase and heat shock protein 90. AB - Adiponectin can protect vessels from injury by promoting the activity of endothelial nitric oxide synthase (eNOS) with increased nitric oxide production. Recently, it was demonstrated that eNOS activity is highly regulated by heat shock protein 90 (HSP90). We tested the hypothesis that adiponectin can prevent endothelial cell injury produced by angiotensin II through promotion of the association between eNOS and HSP90. Cultured human umbilical vein endothelial cells (HUVECs) were treated with angiotensin II (2 microM) to induce apoptosis. In the presence of globular adiponectin, apoptosis was inhibited in a dose response manner. Angiotensin II-induced apoptosis was also inhibited by treatment with an NO donor and by combined treatment with both angiotensin II type 1 and type 2 receptor blockers. Western blotting and immunoprecipitation of the lysates from the treated cells showed that globular adiponectin could restore the association between eNOS and HSP90 and enhance the phosphorylation of eNOS. In conclusion, angiotensin II-induced human endothelial cell apoptosis can be prevented by adiponectin through promotion and stabilization of the association between eNOS and HSP90. PMID- 15358549 TI - Facilitation of proteasomal degradation of p27Kip1 by N-terminal cleavage and their sequence requirements. AB - The sequence requirement for N-terminal cleavage and the proteasomal degradation of p27Kip1 and their relationship was investigated. Residues 5-8 were required for the cleavage and the mutation of S10 to E inhibited the cleavage. The C terminal PEST sequence was necessary for the degradation and residue R165 was found to play an important role in the degradation. The inhibition of the cleavage by deleting residues 5-8 inhibited the degradation, while the fragment mimicking the cleavage product accelerated the degradation. Both the cleavage and degradation demonstrated a similar sensitivity toward proteasome inhibitors and ATP depletion. These two processes are thus suggested to be tightly linked and sequential. PMID- 15358550 TI - Evaluation of metal-conjugated compounds as inhibitors of 3CL protease of SARS CoV. AB - 3C-like (3CL) protease is essential for the life cycle of severe acute respiratory syndrome-coronavirus (SARS-CoV) and therefore represents a key anti viral target. A compound library consisting of 960 commercially available drugs and biologically active substances was screened for inhibition of SARS-CoV 3CL protease. Potent inhibition was achieved using the mercury-containing compounds thimerosal and phenylmercuric acetate, as well as hexachlorophene. As well, 1-10 microM of each compound inhibited viral replication in Vero E6 cell culture. Detailed mechanism studies using a fluorescence-based protease assay demonstrated that the three compounds acted as competitive inhibitors (Ki=0.7, 2.4, and 13.7 microM for phenylmercuric acetate, thimerosal, and hexachlorophene, respectively). A panel of metal ions including Zn2+ and its conjugates were then evaluated for their anti-3CL protease activities. Inhibition was more pronounced using a zinc-conjugated compound (1-hydroxypyridine-2-thione zinc; Ki=0.17 microM) than using the ion alone (Ki=1.1 microM). PMID- 15358551 TI - Atypical protein kinase C stimulates nucleotide excision repair activity. AB - Nucleotide excision repair (NER) deals with bulky DNA damages. However, the regulation of this process is still unclear. Here, we show that both cell resistance to genotoxic agents that generate DNA lesions corrected by NER and in vitro NER activity are correlated with atypical protein kinase C (PKC) zeta expression levels. Moreover, repair intermediates are produced and eliminated more rapidly in UV-irradiated PKCzeta-overexpressing cells. The expression levels of XPC and hHR23B, two NER proteins, are correlated with PKCzeta expression. Altogether, these results strongly suggest that PKCzeta could act as a modulator of NER activity by regulating the expression of XPC/hHR23B heterodimer. PMID- 15358552 TI - The trimeric organisation of photosystem I is not necessary for the iron-stress induced CP43' protein to functionally associate with this reaction centre. AB - A mutant of Synechocystis PCC 6803 lacking the PsaL subunit of photosystem I (PSI) has been grown in iron-deficient media to induce the expression of the isiA gene, which encodes the chlorophyll a-binding protein CP43'. The purpose of this was to establish whether or not the formation of an 18-mer CP43'-PSI supercomplex reported for wild type Synechocystis cells [Nature 412 (2001) 743-745] was dependent on the trimeric conformation of the cyanobacterial PSI reaction centre. Structural characterisation by electron microscopy and single particle image analysis has revealed that the PsaL-mutant does not form trimers of PSI. However, despite this, CP43' was found to associate with the PSI monomer. The PSI monomer bound six or seven copies of CP43' along one edge of the PSI monomer and can be compared with one segment of the trimeric 18-mer CP43'-PSI supercomplex. We therefore conclude that the trimeric nature of cyanobacterial PSI is not required for the assembly of the CP43' antenna system under iron-deficient conditions. PMID- 15358553 TI - Characterization of trans- and cis-cleavage activity of the SARS coronavirus 3CLpro protease: basis for the in vitro screening of anti-SARS drugs. AB - Severe acute respiratory syndrome (SARS) has been globally reported. A novel coronavirus (CoV), SARS-CoV, was identified as the etiological agent of the disease. SARS-CoV 3C-like protease (3CLpro) mediates the proteolytic processing of replicase polypeptides 1a and 1ab into functional proteins, playing an important role in viral replication. In this study, we demonstrated the expression of the SARS-CoV 3CLpro in Escherichia coli and Vero cells, and then characterized the in vitro trans-cleavage and the cell-based cis-cleavage by the 3CLpro. Mutational analysis of the 3CLpro demonstrated the importance of His41, Cys145, and Glu166 in the substrate-binding subsite S1 for keeping the proteolytic activity. In addition, alanine substitution of the cleavage substrates indicated that Gln-(P1) in the substrates mainly determined the cleavage efficiency. Therefore, this study not only established the quantifiable and reliable assay for the in vitro and cell-based measurement of the 3CLpro activity, but also characterized the molecular interaction of the SARS-CoV 3CLpro with the substrates. The results will be useful for the rational development of the anti-SARS drugs. PMID- 15358554 TI - Comparative analysis of phosphoprotein-enriched myocyte proteomes reveals widespread alterations during differentiation. AB - The differentiation of skeletal muscle has been associated with altered phosphorylation status of individual proteins. However, a global analysis of protein phosphorylation during myogenesis has yet to be undertaken. Here, we report the identification of over 130 putative phosphoproteins from murine C2C12 muscle cells. Cell extracts were fractionated on phosphoprotein enrichment columns and the resulting proteins were detected by two-dimensional gel electrophoresis and silver stain, and identified by liquid chromatography coupled to electrospray tandem mass spectrometry. The early differentiation of C2C12 myoblasts was found to be accompanied by changes in the phosphorylation or expression of numerous proteins including cytoskeletal, heat shock and signaling proteins, the pp32 family of nuclear phosphoproteins, several disease-associated gene products and other characterized and uncharacterized proteins. PMID- 15358555 TI - Truncated KCNQ1 mutant, A178fs/105, forms hetero-multimer channel with wild-type causing a dominant-negative suppression due to trafficking defect. AB - We identified a novel mutation Ala178fs/105 missing S3-S6 and C-terminus portions of KCNQ1 channel. Ala178fs/105-KCNQ1 expressed in COS-7 cells demonstrated no current expression. Co-expression with wild-type (WT) revealed a dominant negative effect, which suggests the formation of hetero-multimer by mutant and WT. Confocal laser microscopy displayed intracellular retention of Ala178fs/105 KCNQ1 protein. Co-expression of the mutant and WT also increased intracellular retention of channel protein compared to WT alone. Our findings suggest a novel mechanism for LQT1 that the truncated S1-S2 KCNQ1 mutant forms hetero-multimer and cause a dominant-negative effect due to trafficking defect. PMID- 15358556 TI - RecQ helicase enhances homologous recombination in plants. AB - RecQ helicase is a key component in the RecF pathway of Escherichia coli for initiation of homologous recombination. Here, we demonstrate that transient expression of RecQ gene in rice embryogenic cell increases the homologous recombination efficiency as much as 4-fold. Further experiments reveal that this effect is influenced by the RecQ dosage. Stable expression of RecQ in rice dramatically increases the homologous recombination events 20- to 40-fold in leaf tissue from different transgenic lines. This is the first evidence indicating that overexpression of RecQ gene can stimulate homologous recombination in plants. PMID- 15358557 TI - Exon 6 of human Jagged-1 encodes an autonomously folding unit. AB - Human Jagged-1 is predicted to contain 16 epidermal growth factor-like (EGF) repeats. The oxidative folding of EGF-2, despite the several conditions tested, systematically led to complex mixtures. A longer peptide spanning the C-terminal part of EGF-1 and the complete EGF-2 repeat, on the contrary, could be readily refolded. This peptide, which corresponds to the entire exon 6 of the Jagged-1 gene, thus represents an autonomously folding unit. We show that it is structured in solution, as suggested by circular dichroism and NMR spectroscopy, and displays an EGF-like disulfide bond topology, as determined by disulfide mapping. PMID- 15358558 TI - Mutational analysis of conserved AAA+ residues in the archaeal Lon protease from Thermoplasma acidophilum. AB - The Lon protease from the archaeon Thermoplasma acidophilum (TaLon) is composed of an N-terminal ATPase associated with various cellular activities (AAA+) domain and a C-terminal Lon protease domain. Although related in sequence to the soluble Lon proteases, TaLon was shown to be membrane-bound in its native host and also when expressed in Escherichia coli. Recombinant TaLon was purified as a functional high-molecular weight complex displaying ATPase and proteolytic activity. Mutagenesis of conserved AAA+ residues revealed that the Walker A and B motifs, and the sensor 1 and sensor 2' residues were essential for the ATPase activity, while the sensor 2 and the arginine finger were involved in activation of the protease domain. PMID- 15358559 TI - Evolutionary analysis of 11beta-hydroxysteroid dehydrogenase-type 1, -type 2, type 3 and 17beta-hydroxysteroid dehydrogenase-type 2 in fish. AB - Steroid dehydrogenases regulate the access of active steroids to their receptors. In particular, 11beta-hydroxysteroid dehydrogenase-type 1 (11beta-HSD1) and 11beta-HSD2 regulate the levels of glucocorticoids, such as cortisol, and 17beta HSD1 and 17beta-HSD2 regulate the levels of androgens and estrogens. Human 11beta HSD1 and 11beta-HSD2 are distant homologs, with less than 25% amino acid sequence identity, as are human 17beta-HSD1 and 17beta-HSD2. In contrast, human 11beta HSD2 and 17beta-HSD2 are close homologs, with about 43% sequence identity. Until recently, deciphering early events in the evolution of 11beta-HSD2 and 17beta HSD2 was difficult because only mammalian sequences were available. The completely sequenced Takifugu, Tetraodon and medaka genomes and the almost completed zebrafish genome provide an opportunity to investigate the evolution of 11beta-HSD2, 17beta-HSD2, and 11beta-HSD1. Unexpectedly, a search of the Takifugu, Tetraodon and medaka genomes only found an ortholog to 11beta-HSD2 and none to 17beta-HSD2, while the zebrafish genome contains orthologs of both enzymes. This suggests that 17beta-HSD2 was lost in teleosts after the divergence of zebrafish and medaka. Also unexpectedly, searches with 11beta-HSD1 only identified several fish 11beta-HSD3s, as well as an ortholog in Ciona, indicating that 11beta-HSD3 is the ancestor of 11beta-HSD1. PMID- 15358560 TI - Trichostatin A activates p18INK4c gene: differential activation and cooperation with p19INK4d gene. AB - We have reported that histone deacetylase (HDAC) inhibitors activate a member of the INK4 family, the p19INK4d gene, causing G1 phase arrest. We report here that HDAC inhibitor, Trichostatin A, activates another member of the INK4 family, the p18INK4c gene, through its promoter in Jurkat cells. Interestingly, the activation patterns of the p18INK4c gene were different from those of p19INK4d. Furthermore, mouse embryo fibroblasts lacking p18Ink4c or p18Ink4c/p19Ink4d were resistant to the growth inhibitory effects of TSA as compared to their wild-type counterpart. Our findings suggest that p18INK4c is involved in TSA-mediated cell growth inhibition and cooperates with p19INK4d. PMID- 15358561 TI - Cyanidin-3-glucoside regulates phosphorylation of endothelial nitric oxide synthase. AB - Dietary anthocyanins are widely distributed in fruits, vegetables and red wines, and there are several reports mentioning their beneficial effects. Previously we reported that cyanidin-3-glucoside (Cy3G) induced endothelial nitric oxide synthase (eNOS) expression and nitric oxide (NO) production in bovine vascular endothelial cells. In the present study, we show that Cy3G regulates phosphorylation of eNOS and Akt, affects the interaction between eNOS and soluble guanylyl cyclase, and increases cyclic guanosine monophosphate (cGMP) production. Our results suggest that Cy3G enhances vascular eNOS activity, and may help to improve vascular endothelial function. PMID- 15358562 TI - Heat shock protein-90 dampens and directs signaling stimulated by insulin-like growth factor-1 and insulin. AB - Heat shock protein-90 (Hsp90) buffers cells from genetic mutations and environmental stresses. To test if this capability reflects a normal physiological function of Hsp90 to buffer cellular signals, the effects of Hsp90 inhibition were measured on activation of Akt. Inhibition of Hsp90 with geldanamycin amplified Akt phosphorylation induced by insulin-like growth factor 1 (IGF-1) or insulin, indicating that Hsp90 normally buffers these signals. Furthermore, with IGF-1 stimulation Hsp90 inhibition increased p38 activation, produced additive activation of p90RSK, and slightly increased the duration of ERK1/2 activation. Hsp90 dampened Akt signaling by facilitating phosphatase mediated dephosphorylation of Akt. Thus, Hsp90 not only buffers the cellular effects of mutations and stresses, but also buffers the magnitude and duration of activation of proliferative and survival-promoting signaling responses. PMID- 15358563 TI - Transferrin receptor 1 mRNA is downregulated in placenta of hepcidin transgenic embryos. AB - We have previously shown that hepcidin transgenic embryos are severely anemic and die around birth. Here, we report that embryonic hepcidin transgene expression decreases transferrin receptor 1 (TfR1) mRNA level in placenta, as shown by cDNA microarray analysis and quantitative RT-PCR, by a mechanism which is independent of placenta iron content and iron responsive element/iron regulatory protein (IRE/IRP) activity. On the contrary, iron injections into pregnant mothers result in increased placenta iron and ferritin content, and reduced IRE binding activity of IRP1 leading to decreased TfR1 mRNA level. Taken together, these results suggest that hepcidin action on placenta is mostly through transcriptional downregulation of the iron uptake machinery. PMID- 15358564 TI - Genome based identification and analysis of the pre-replicative complex of Arabidopsis thaliana. AB - Eukaryotic DNA replication requires an ordered and regulated machinery to control G1/S transition. The formation of the pre-replicative complex (pre-RC) is a key step involved in licensing DNA for replication. Here, we identify all putative components of the full pre-RC in the genome of the model plant Arabidopsis thaliana. Different from the other eukaryotes, Arabidopsis houses in its genome two putative homologs of ORC1, CDC6 and CDT1. Two mRNA variants of AtORC4 subunit, with different temporal expression patterns, were also identified. Two hybrid binary interaction assays suggest a primary architectural organization of the Arabidopsis ORC, in which AtORC3 plays a central role in maintaining the complex associations. Expression profiles differ among pre-RC components suggesting the existence of various forms of the complex, possibly playing different roles during development. In addition, the expression of the putative pre-RC genes in non-proliferating plant tissues suggests that they might have roles in processes other than DNA replication licensing. PMID- 15358565 TI - Preparation of artificial 2-, 3-, 4- and 8-domain myoglobins and comparison of their autoxidation rates. AB - Although most hemoglobins and myoglobins consist of 15-kDa single-domain subunits, structurally unusual hemoglobins, such as Artemia 9-domain and Barbatia 2-domain hemoglobins, occur naturally in several invertebrates. These hemoglobins appear to be the result of gene duplication and fusion. Using cDNA coding for the open reading frame of Aplysia kurodai myoglobin, artificial cDNA inserts corresponding to contiguous dimer, trimer, tetramer and octamer myoglobins (2-, 3 , 4- and 8-domain myoglobins) were prepared and cloned into pMAL or pQE plasmids. These artificial myoglobins and wild-type single-domain myoglobins were successfully expressed in Escherichia coli in the heme-attached, oxygenated form. Myoglobin was purified partially by ammonium sulfate fractionation and gel filtration, and autoxidation rates were examined. The autoxidation rates of recombinant wild-type myoglobins with MBP or hexameric His tag were comparable to those of native myoglobin, suggesting that the recombinant proteins appear to be properly folded and that the N-terminal MBP or His tag does not have an affect on the rate. On the other hand, the rates were significantly decreased in the 2- and 3-domain myoglobins (50% and 30% of the single-domain myoglobins, respectively). The rates for 4- and 8-domain myoglobins were similar to those for 3-domain myoglobin. These results indicate that the artificial poly-domain structure of myoglobin is more stable than the usual single-domain myoglobin from the viewpoint of storage of bound dioxygen. PMID- 15358566 TI - Allergic contact dermatitis. AB - Contact dermatitis is an inflammatory skin condition induced by exposure to an environmental agent. Eczema and dermatitis are used synonymously to denote a polymorphous pattern of skin inflammation characterized at least in its acute phase by erythema, vesiculation and pruritus. Substances responsible for contact dermatitis after single or multiple exposures are non protein chemicals, i.e. haptens, that induce skin inflammation through activation of innate skin immunity (irritant contact dermatitis) or both innate and acquired specific immunity (allergic contact dermatitis). The present review will focus on allergic contact dermatitis, a delayed-type hypersensitivity reaction, which is mediated by hapten specific T cells. Recent advances in the pathophysiology of ACD have shown that the occurrence of ACD, as well as its magnitude and duration, is controlled by the opposite functions of CD8 effector T cells and CD4 regulatory T cells. From these studies ACD can be considered as a breakdown of cutaneous immune tolerance to haptens. PMID- 15358567 TI - Dermatological manifestations of Lyme borreliosis. AB - Lyme borreliosis is a multisystem infectious disease caused by the tick transmitted spirochete Borrelia burgdorferi sensu lato. About 80% of all Lyme borreliosis cases represent skin manifestations (dermatoborrelioses). The three characteristic dermatoborrelioses are erythema migrans, borrelial lymphocytoma, and acrodermatitis chronica atrophicans, which occur in different stages of the disease. Erythema migrans is the hallmark of early Lyme borreliosis, whereas acrodermatitis chronica atrophicans is the characteristic manifestation of late Lyme borreliosis. Several spirochetal factors (e.g. infection with different genospecies, co-infection with other tick-transmitted pathogens) as well as host factors (e.g. cytokine patterns at the site of infection) influence the course of the disease. Diagnosis in the early stage of Lyme borreliosis relies on the clinical picture, whereas serological, molecular, microbiological, and histopathological findings are important adjuncts in the diagnosis of later stages of the infection. Antibiotic treatment is necessary for all stages and manifestations of Lyme borreliosis. Doxycycline is the antibiotic of choice for most patients with dermatoborrelioses. PMID- 15358568 TI - Hyalinosis cutis et mucosae: a 30 year follow up of a female patient. AB - Hyalinosis cutis et mucosae is an inherited disease, characterized by deposition of a hyaline-like substance in the dermis and internal organs. In the present study, we report a 30 year follow up in a female patient with hyalinosis cutis et mucosae treated using dermabrasion and oral retinoids. In our opinion, the lesion on uncovered body areas should be corrected as soon as possible to enable a normal psychophysical development. The lesions on skin and oral mucosa may be successfully removed without remission with dermabrasion in early childhood. In addition, oral retinoids in adulthood may produce some improvement. PMID- 15358569 TI - Nevus psiloliparus: report of two nonsyndromic cases. AB - A diagnosis of nevus psiloliparus was made both clinically and histopathologically in two otherwise healthy girls, one being 4 years and the other one being 1 year old. A congenital hairless patch with a round or oblong shape and a soft surface was noted on the scalp. In one case the lesion was yellowish and flat, whereas in the other case it was skin colored and somewhat elevated. In both cases, histopathological examination showed the absence of mature hair follicles and the presence of undeveloped follicular structures as well as orphaned arrector pili muscles in the dermis. As a new histopathological feature of this type of nevus, we found arrested anlagen of hair bulbs in both cases. The fatty tissue was abundant and also involved, in the form of aberrant lobules, the lower portion of the dermis. Clinical examination did not show any associated extracutaneous abnormality, and during a follow-up period of 2 years in either case, respectively, the children developed without any complication. Although neurological abnormalities could not be excluded by imaging techniques, such extracutaneous involvement is highly unlikely because the nevi psilolipari were of rather limited size. When clinicians and dermatohistopathologists have become familiar with this new entity, they will most likely recognize it as a nonsyndromic skin disorder more often than as a cutaneous sign of encephalocraniocutaneous lipomatosis. PMID- 15358570 TI - CD4+ CD56+ blastic tumor of the skin: cytogenetic observations and further evidence of an origin from plasmocytoid dendritic cells. AB - We here report on a case of a blastic tumor, recently described to belong to a new entity sharing phenotypic similarities with blood derived plasmocytoid dendritic cells and formerly regarded as belonging to the group of natural killer cell lymphomas. Besides immunophenotypic characteristics such as the absence of T cell markers and almost complete absence of markers of the myeloid lineage, these tumors express CD4, CD56 and CD123, the receptor for interleukin-3. Moreover, using the comparative genomic hybridisation technique, CGH, we demonstrate a gain of chromosome 7q, 22 and a loss of chromosome 3p and 13q. Since this type of hematologic disorder often shows its primary manifestation in the skin and often runs a rapidly fatal course, it is important to distinguish this from other types of primary cutaneous lymphomas. PMID- 15358571 TI - Effects of local corticosteroids on acute experimental urticaria. AB - Corticosteroids are often used in the treatment of acute or chronic urticaria. However, their effects on mastocyte activation as well as on the histamine induced dermal oedema remain poorly investigated. The aim of the present study was to investigate the effects of corticosteroids (CS) on the development of acute experimental urticaria induced by prick-tests with histamine and codeine. This experimental model corresponds to the common form of urticaria. CS were administered at the site of the histamine and codeine prick tests in order to test for a direct effect on the development of acute urticaria. Two types of experiments were performed: 1) after a 48-hour period of topical CS application on the forearm, 7 healthy volunteers were skin prick-tested with histamine and codeine simultaneously in duplicate, one series in the pretreated area and the other in a non-treated area. 2) six other volunteers were prick-tested with histamine and codeine on their forearm, in duplicate. Immediately after testing, intradermal methyprednisolone was injected at the site of the prick-tests in the last series. Skin wheal and flare responses were measured after 20 mns and statistically compared with and without CS treatment. Whereas short-term CS topical application did not appear to modify cutaneous reactivity to histamine and codeine, local CS injection was associated with a significant increase in the flare induced by histamine and codeine (respectively + 18 +/- 3% and + 38 +/- 3%; P = 0.05). The wheal tended to be increased after injected CS. In conclusion, these results show that CS are neither able to prevent nor to improve experimental urticaria, i.e. wheal and flare, and even increase the histamine and codeine-induced erythema. That a similar result could apply to patients with chronic urticaria and with systemic CS remains to be studied. PMID- 15358572 TI - Chromosome aberrations in Raynaud's phenomenon. AB - We evaluated the occurrence of spontaneous chromosome damage in cultured peripheral lymphocytes of subjects with idiopathic and pre-scleroderma Raynaud's phenomenon, by means of molecular cytogenetic analysis. Using the micronucleus assay as a marker of chromosome alteration, we studied 30 patients with pre scleroderma Raynaud's phenomenon, 30 patients with idiopathic Raynaud's phenomenon and 30 healthy subjects. All subjects were classified as ANA-, ACA+ or Scl 70+. To identify the mechanism of micronucleus formation, fluorescence in situ hybridisation analysis was also performed. Pre-scleroderma Raynaud's phenomenon subjects showed significantly higher micronucleus frequencies than idiopathic Raynaud's phenomenon subjects and controls (37.0 +/- 11.5 vs. 11.1 +/- 3.2 and 10.7 +/- 2.7 respectively p < 0.0001). Interestingly, subjects with idiopathic Raynaud's phenomenon displayed micronucleus frequency comparable to that of healthy controls. Furthermore, ACA+ subjects showed the highest micronucleus frequencies (41.0 +/- 7.6) as compared to subjects with Scl 70+ antibody (25.0 +/- 3.5). Our results show that circulating lymphocytes of only pre-scleroderma Raynaud's phenomenon subjects undergo chromosomal damage, as detected by the micronucleus assay, at a higher rate than expected. No prevalence of aneuploidogenic or clastogenic events in micronucleus formation is revealed by fluorescence in situ hybridisation analysis. PMID- 15358573 TI - Treatment of alopecia areata in the DEBR model using Cyclosporin A lipid vesicles. AB - Alopecia areata (AA) is a chronic cutaneous disease with a suspected autoimmune origin. We evaluated the efficacy of 0.5% Cyclosporin A (CyA) in a topically applied liposomal formulation as a potential treatment for AA using the Dundee Experimental Bald Rat (DEBR) model. The vehicle consisted of liposomes (75% phosphatidylcholine, 5% lysophosphatidylcholine, 5% sterol, natural oils) of 10% wt. in ethanol with and without 2% wt. terpenes (d-limonene: citral: cineole, 10:45:45) as a penetration enhancer (PE). Fifteen DEBR were allocated to 3 groups of 5. Groups I, II and III received CyA vesicles with PE, CyA vesicles without PE, and CyA in ethanol respectively. All rats were treated twice a day for 6 weeks within a 4 cm2 area on one bald flank with CyA while the contralateral flank received an equivalent control formulation. Rats in group I exhibited visible hair regrowth on the drug treated site after one week of drug application. Group II rats had visible hair regrowth by the end of the second week. The hair growth was progressive and reached a maximum density at the site of application after six weeks in both groups. Histological examination revealed a reduced inflammatory infiltrate and improved hair follicle morphology within the drug treated area as compared to the contralateral vehicle treated skin. Group III rats showed neither visible signs of hair growth nor reduction of hair follicle inflammation. The results of this proof of concept preliminary study suggest that CyA vesicle formulations with and without PE have promising potential as a topical treatment for AA in humans. PMID- 15358574 TI - DRESS syndrome associated with carbamazepine and phenytoin. AB - Drug Rash with Eosinophilia and Systemic Symptoms (DRESS) syndrome reflects a serious hypersensitivity reaction to drugs. Its clinical manifestations include diffuse maculopapular rash, exfoliative dermatitis, facial edema, lymphadenopathy, fever, multivisceral involvement and it is associated with a high mortality rate. We report a 62-year-old patient suffering from epilepsia presenting erythroderma following carbamazepine intake. Blood tests revealed eosinophilia, leukocytosis, elevated liver enzymes and high levels of Eosinophil Cationic Protein (ECP). We applied systemic steroids and anticonvulsant therapy was switched to phenytoin, which had been taken previously without adverse reactions. The skin eruptions persisted and the patient developed fever. Anticonvulsant medication was discontinued and skin eruptions finally resolved under steroid application. This case report demonstrates that cross reactivity between carbamazepine and phenytoin may not only lead to the development but also to the worsening of DRESS syndrome. ECP blood levels may represent a sufficient parameter to monitor the development of DRESS syndrome. PMID- 15358575 TI - Cutaneous ulcerations on hands and heels secondary to long-term hydroxyurea treatment. AB - Hydroxyurea is a chemotherapeutic agent used to treat myeloproliferative disorders and other non-neoplastic conditions. Cutaneous side-effects have been described in long-term therapy with hydroxyurea and include xerosis, hyperpigmentation, skin atrophy, erythema, alopecia, skin tumours and ulceration of the skin, particularly of the legs. We present a 71-year old patient with chronic myelocytic leukemia (CML) who developed extremely painful ulcers on the hands and heels as well as skin tumours while on long-term therapy with hydroxyurea. The ulcers were resistant to therapy but healed three months after discontinuation of hydroxyurea therapy. PMID- 15358576 TI - Ciliated cyst of the vulva. AB - Ciliated cyst of the vulva is a rare anomaly. The cyst is lined with columnar ciliated epithelium that resembles mullerian epithelium and is located on the superior portion of the labium minus, occasionally associated with pregnancy or exogenous progesterone. We describe a case of an 11-year-old Japanese girl with a ciliated cyst of the vulva and a history of atopic dermatitis. Epithelial cells in our case showed negative staining for estrogen receptor. The origin of the ciliated epithelium is still unknown. However, chronic inflammation of the vulvar vestibulae may contribute to the pathogenesis of ciliated cyst of the vulva. PMID- 15358577 TI - Chronic external iliac vein obstruction as a cause of leg ulceration. AB - Chronic venous leg ulceration, which tends to recur, is difficult to treat and therefore needs special diagnostic and therapeutic care. We recently treated a 45 year-old patient with an obstruction of the left external iliac vein, caused by deep venous thrombosis. We would like to propose that, although rare, the existence of pelvic vein thrombosis and obstruction can cause venous leg ulcers. This case clearly pointed out that in patients with crural leg ulceration, showing no other signs of chronic venous insufficiency and where duplex ultrasound is normal, additional diagnostic evaluation should be performed. Because standard duplex ultrasound investigation can fail to demonstrate the obstruction, phlebography should be used in suspected cases as a secondary test to check both the superficial and the deep venous system for pathology, applying the endovascular therapeutic stent-placement technique. PMID- 15358578 TI - Nodular cystic fat necrosis with systemic sclerosis. AB - Nodular cystic fat necrosis is a distinct spectrum characterized clinically by mobile subcutaneous nodules and histologically by encapsulated fat necrosis. We describe herein a case of nodular cystic fat necrosis in a patient with systemic sclerosis in the atrophic stage. Several mobile, firm nodules were surgically removed from the flexural aspect of the forearm and lower leg. Histopathology showed features of nodular cystic fat necrosis with lipomembranous changes and calcification. Of interest, lipomembranous changes were seen also in the biopsied specimen from the extensor aspect of the forearm of scleroderma. We speculate that multiple, chronic, local or systemic events causing a compromise in the blood supply of the subcutaneous tissues may contribute to the induction of lipomembranous changes in the affected skin as well as nodular cystic fat necrosis in this case. PMID- 15358579 TI - The mammalian beta globin origin of DNA replication. AB - Initiation of DNA replication is a tightly regulated process aimed to insure that the entire genome is replicated at the appropriate time during each cell cycle. In the human beta globin locus, replication initiates from a region between the two genes that encode the adult subunit of hemoglobin (the beta globin initiation region, or IR). Mammalian beta globin loci replicate early during the S phase of the cell cycle in pre erythroid cells, in which the beta-globin locus is present in a euchromatin form. However, in cells that do not express globin and in which the locus is heterochromatic, these same loci replicate during the later stages of S phase. Both early and late replication patterns utilize similar replication initiation regions. These features make the beta globin locus an attractive model for studying the determinants of replication sites and replication timing, as well as the correlation between gene expression and DNA replication. Two genomic domains are essential for initiation of DNA replication within the locus: the initiation region (IR), and a 40 kb region upstream of the globin gene cluster known as the locus control region (LCR). The IR meets the genetic requirements for a chromosomal replicator, since it can initiate DNA replication at ectopic sites. The LCR regulates transcriptional activity and chromatin structure, and may act as a determinant of replication timing. This review will summarize recent findings characterizing the sequence requirements for initiation of DNA replication in mammalian beta globin loci and will discuss the specific influence of the location and the chromosomal environment in regulating DNA replication at the beta globin IR. PMID- 15358580 TI - Dual functional regulators coordinate DNA replication and gene expression in proliferating cells. AB - Gene products for cell growth must meet the pace of DNA replication and vice versa during the cell division cycle, therefore coordination of DNA replication and gene expression is vital to proliferating cells. During development in multicellular organisms when rapid cell divisions must be accompanied by the expression of particular gene sets in differentiating tissues, this coordination is even more crucial. Undoubtedly, multiple strategies are used to ensure the coordination of gene expression and DNA replication. In this review, we focus on the strategy that uses dual functional factors to serve both the functions of replication initiator and transcription regulator. Classical examples are the dual functional replication initiator/transcription regulators, DnaA of E. coli and T antigen of SV40, which bind replication origins and regulate their own synthesis. Emerging examples in eukaryotes are the growth responsive transcription factor E2f, the MADS domain combinatorial transcription factor Mcm1, and a subunit of the MCM2-7 helicase, Mcm7. PMID- 15358582 TI - Functional roles of NKT cell in the immune system. AB - CD1d-restricted Natural Killer T cells (NKT cells), a novel lymphocyte lineage, are considered to play an intermediary role bridging innate and acquired immunity. This review discusses the characteristics of NKT cells and their biological significance in the immune system, and summarizes their in vivo functions observed in a number of pathological settings, including infectious diseases, cancer, autoimmunity, and transplantation. Further, we discuss recent data that have generated considerable interest in utilizing NKT cells as targets of new therapeutic interventions in various human diseases. PMID- 15358581 TI - Role of accessory proteins of HTLV-1 in viral replication, T cell activation, and cellular gene expression. AB - Human T-cell lymphotropic virus type 1 (HTLV-1), causes adult T cell leukemia/lymphoma (ATLL), and initiates a variety of immune mediated disorders. The viral genome encodes common structural and enzymatic proteins characteristic of all retroviruses and utilizes alternative splicing and alternate codon usage to make several regulatory and accessory proteins encoded in the pX region (pX ORF I to IV). Recent studies indicate that the accessory proteins p12I, p27I, p13II, and p30II, encoded by pX ORF I and II, contribute to viral replication and the ability of the virus to maintain typical in vivo expression levels. Proviral clones that are mutated in either pX ORF I or II, while fully competent in cell culture, are severely limited in their replicative capacity in a rabbit model. These HTLV-1 accessory proteins are critical for establishment of viral infectivity, enhance T-lymphocyte activation and potentially alter gene transcription and mitochondrial function. HTLV-1 pX ORF I expression is critical to the viral infectivity in resting primary lymphocytes suggesting a role for the calcineurin-binding protein p12I in lymphocyte activation. The endoplasmic reticulum and cis-Golgi localizing p12I activates NFAT, a key T cell transcription factor, through calcium-mediated signaling pathways and may lower the threshold of lymphocyte activation via the JAK/STAT pathway. In contrast p30II localizes to the nucleus and represses viral promoter activity, but may regulate cellular gene expression through p300/CBP or related co-activators of transcription. The mitochondrial localizing p13II induces morphologic changes in the organelle and may influence energy metabolism infected cells. Future studies of the molecular details HTLV-1 "accessory" proteins interactions will provide important new directions for investigations of HTLV-1 and related viruses associated with lymphoproliferative diseases. Thus, the accessory proteins of HTLV-1, once thought to be dispensable for viral replication, have proven to be directly involved in viral spread in vivo and represent potential targets for therapeutic intervention against HTLV-1 infection and disease. PMID- 15358583 TI - Role of gammadelta T lymphocytes in tumor defense. AB - The effectors of mucosal and natural immunity (i.e. natural killer, NK, cells and NKT lymphocytes) are known to play an important role in host defence against tumors. Gammadelta T lymphocytes are the most represented cell populations in mucosal associated lymphoid tissue and share several characteristics of T and NK cells. Two main subsets of gammadelta T cells are known: one, expressing the Vdelta2 T cell receptor (TCR), is found in the peripheral blood, while T cells expressing Vdelta1 TCR are resident in epithelial tissues. The former subset is capable of killing myeloma and Burkitt lymphoma cells, while the latter has been implied in the defence against epithelial cancers. Furthermore, there is increasing evidence that alphabeta and gammadelta T lymphocytes make distinct contributions to anticancer surveillance. Indeed, unlike alphabeta T cells, gammadelta T lymphocytes are involved in the recognition of antigens that do not undergo the conventional major histocompatibility complex (MHC)-driven antigen presentation. Down-regulation of expression of MHC alleles as well as tumor specific antigens is observed frequently during tumor progression, resulting in an impairment of MHC-restricted, alphabeta-T-cell-mediated tumor-specific immunity. Given the unique set of antigens recognized and the lack of requirement for classical antigen-presenting molecules, gammadelta T lymphocytes might, therefore, represent a useful and potent system in anti-cancer surveillance, as proposed for the immune response against pathogens. Evidence that gammadelta and alphabeta cells make distinct contribution to anti-cancer surveillance have been recently provided in mice. Here, we discuss the potential role played by resident Vdelta1+ and circulating Vdelta2+ T lymphocytes in the defense against solid tumors and hematological malignancies. PMID- 15358584 TI - Beta-lactams and their potential use as novel anticancer chemotherapeutics drugs. AB - The discovery of natural and synthetic antibiotics is one of the most important medical breakthroughs in human history. Many diseases, such as bacterial meningitis, pneumonia, and septicemia, are now curable with the use of antibiotics. Antibiotics are efficacious, generally well tolerated in patients, and have a low toxicity level. It is for these reasons antibiotics remain an attractive target for drug discovery. Traditional beta-lactam antibiotics (e.g. penicillins, penems, cephalosporins) have a bicyclic ring structure that is conformationally rigid and functions to inhibit bacterial cell wall synthesis. In addition to the bactericidal action of antibiotics, it has been discovered that many antibiotics are capable of inhibiting tumor cell growth. There are currently many antitumor antibiotics approved for cancer therapy, which work to inhibit tumor cell growth by DNA intercalation. The use of beta-lactams as prodrugs has also met with success by aiding delivery of the chemotherapeutic directly to tumor sites. Recently, a novel class of N-thiolated monobactams, so termed because they possess a monocyclic ring instead of the bicyclic ring, has been found to induce apoptosis potently and specifically in many tumor cell lines but not in normal, non-transformed cell lines. Other beta-lactams, such as the polyaromatics, have been found to slow or inhibit tumor cell growth, and the 4 alkylidene beta-lactams are capable of inhibiting matrix metalloproteinases and leukocyte elactase activity. These data indicate that synthesis and evaluation of beta-lactams are a promising area for further development in anticancer research. PMID- 15358585 TI - Green tea and tea polyphenols in cancer prevention. AB - The cancer-preventive effects of green tea and its main constituent (-) epigallocatechin gallate [(-)-EGCG] are widely supported by results from epidemiological, cell culture, animal and clinical studies in the recent decade. In vitro cell culture studies show that tea polyphenols potently induce apoptotic cell death and cell cycle arrest in tumor cells but not in their normal cell counterparts. Green tea polyphenols affect several signal transduction pathways, including growth factor-mediated, the mitogen-activated protein kinase (MAPK) dependent, and ubiquitin/proteasome degradation pathways. Epidemiological studies have suggested that the consumption of green tea lowers the risk of cancer. Various animal studies have revealed that treatment by green tea inhibits tumor incidence and multiplicity in different organ sites such as skin, lung, liver, stomach, mammary gland and colon. Phase I and II clinical trials were carried out recently to explore the anticancer effects of green tea in patients with cancer. At this time, more mechanistic research, animal studies, and clinical trials are necessary to further evaluate the role of green tea in cancer prevention. PMID- 15358586 TI - Sphingolipids in the chemoprevention of colon cancer. AB - Sphingolipids were first described more than 100 years ago by the physician Thudicum who named the fatty substance he found in brain, sphingosin. Among multiple other functions, sphingolipids are lipid messengers in the signaling pathways of growth factors, cytokines, cellular stresses and others. As such, they are involved in the regulation of a wide spectrum of processes that modulate cell growth and cell death. These functions may be beneficial in cancer cells that escape growth regulation and exhibit unlimited proliferation. The effects of exogenous sphingolipids on cancer cells in vitro have been well documented; however, the effects of sphingolipids in vivo are less well understood. Since the mechanisms sphingolipids utilize in the prevention of cancer may be different from those in cancer treatment, modulation of cell growth versus induction of cell death, this review will focus on the known effects of orally administered sphingolipids in the prevention of colon cancer in different rodent models, and discuss the effect of sphingolipid metabolites on changes in cell proliferation and cell death that are important events in early carcinogenesis. PMID- 15358587 TI - Regulation of skin cell homeostasis by gamma delta T cells. AB - Although innate T lymphocytes such as gamma delta T cells have been extensively studied, their biological role has remained an enigma to researchers for many years. However, recent advances have begun to explain their complex role in the immune system. Gamma delta T cells are often the major T cell population in epithelial tissues such as the skin, gut, and lung where they have been implicated in maintaining tissue integrity, defending against pathogens, and regulating inflammation. The gamma delta T cells that reside in the skin are a prototypical intra-epithelial lymphocyte (IEL) population. These skin gamma delta T cell receptor (TCR)-expressing cells are named dendritic epidermal T cells (DETC) for their unique dendritic morphology. Using their gamma delta TCR, DETC recognize an unknown ligand expressed by stressed or damaged keratinocytes. Activated DETC exhibit effector functions such as cytokine production, cytolysis, and proliferation in vitro. Recent findings have shown that upon activation by damaged keratinocytes, DETC produce a key keratinocyte growth factor for wound repair called fibroblast growth factor 7 (FGF-7). FGF-7 is produced in vitro and in vivo, suggesting that DETC might play an important role in the biological function of wound repair. Indeed a delay in wound closure and a decrease in the proliferation of keratinocytes at the wound site have been observed in mice lacking gamma delta T cells. In addition to effector functions attributed to DETC, it has also been suggested that gamma delta T cells such as DETC have regulatory roles such as initiating or inhibiting inflammation. This is supported by the findings that DETC produce chemokines and cytokines. Control of the inflammatory response in the epithelium may provide another mechanism to reestablish homeostasis after a biological insult such as wound infliction. Understanding the function of DETC may be useful in the development of future therapies for chronic wounds and the maintenance of skin homeostasis. PMID- 15358588 TI - Copper storage diseases: Menkes, Wilsons, and cancer. AB - The trace element copper is vital to the healthy functioning of organisms. Copper is used in a multitude of cellular activities including respiration, angiogenesis, and immune responses. Like other metals, copper homeostasis is a tightly regulated process. Copper is transported from dietary intake through the serum and into cells via a variety of transporters. There are a variety of copper chaperones designed to insure that copper is sequestered from interaction with cellular membranes, proteins, or DNA where its properties can result in oxidative damage. However, there are disease states in which copper transporters crucial to homeostasis are impaired resulting in potentially toxic copper accumulation. Wilsons and Menkes diseases are two such cases. Wilsons disease (hepatolenticular degeneration) is an autosomal recessive disorder resulting in extreme accumulation of copper in the liver with deposits elsewhere in the body. Menkes is characterized by a systemic copper deficiency (different from the liver specificity of Wilsons disease) and is the result of an X-linked recessive mutation in a copper transporter. Uptake of copper is impaired due to inability to remove existing copper from cells primarily in the small intestine. Though the causes are dramatically different, cancer also shares a similar diagnostic in the accumulation of copper in effected tissues. Studies have shown greatly elevated levels of copper in cancer tissues, and some diagnostics and treatments from Wilsons and Menkes diseases, such as copper chelation therapy, have been used in the treatment of cancer. Given the commonality of copper accumulation in these diseases and that common therapies exist between them, it may prove beneficial to study all three diseases in light of copper homeostasis. This review will examine the chemical nature and biological roles of copper, Wilsons and Menkes disease and their therapies, and the use of copper related therapies in cancer. PMID- 15358589 TI - Molecular mechanisms of chemoprevention and therapy of cancer by retinoids. AB - Chemoprevention is the use of noncytotoxic therapeutic intervention at the early stages of carcinogenesis against the development and progression of mutant clones to invasive cancer. Retinoids are the most extensively studied and one of the most prominent groups of chemopreventive agents to reach clinical trials. Acute promyelocytic leukemia is the first human malignancy that is successfully treated with all-trans retinoic acid. The t(15;17)(q22;q21) gene rearrangement and PML/RARalpha fusion product in acute promyelocytic leukemia played the key role to leukemogenesis and to sensitivity to differentiation-inducing therapy of all trans retinoic acid. This review focuses on retinoid-based chemoprevention and therapy of cancer, and use acute promyelocytic leukemia as a model to illustrate the molecular mechanisms of retinoid signaling pathway. PMID- 15358590 TI - Genotypic and phenotypic biomarker profiles for individual risk assessment and cancer detection (lessons from bladder cancer risk assessment in symptomatic patients and workers exposed to benzidine). AB - There is a need for improved methods for detecting individuals at risk for cancer to target subsets of patients for more intensive individual screening and targeted cancer therapy and chemoprevention. One approach for accomplishing this objective is to detect premalignant molecular fingerprints in an organ at risk for cancer or to define biomarkers reflective of treatment selection and response. Bladder cancer is an excellent model for testing this approach; however, comprehending the strategy for biomarker selection and analysis is more complicated than is generally appreciated. The objective of this article is to provide a succinct overview of our experience with the selection of biomarkers for bladder cancer detection, first in symptomatic patients and then in high-risk cohorts of workers at risk for bladder cancer. Biomarker selection depends on multiple parameters, each of which must be optimized to enhance the utility of a biomarker for clinical application. Many markers that initially show promise fail in the clinical arena for a variety of reasons. Important parameters include when a biomarker is expressed in carcinogenesis (i.e., early vs. late), the sample type, and the method of analysis. These all contribute to the sensitivity, specificity, and ultimate clinical utility of a biomarker. New technologies/ support the notion that all diseases start in the cell, and Seymore West indicated the cell, under appropriate conditions, can function as a microcuvette for biophysical cytochemical analysis. Spectroscopy provides an accurate and sensitive method for quantitative single-cell proteomics. Improved and more stable fluorescence probes will enhance the utility of cellular chemistry, as will a rationale approach for biomarker selection based on the concepts of field cancerization, complemented by improved quantitative analysis of protein markers at the single-cell level. Our laboratory has developed a platform for single-cell proteomic analysis that can be applied to multiple basic science and clinical problems. Single-cell proteomics also facilitates the study of genetic instability and epigenetic signaling (stromal-epithelial interactions) in relation to cancer therapy and diagnosis. Because most cancers arise through multiple signaling pathways and are heterogeneous, the identification of appropriate biomarker profiles provides a number of strategic advantages over a single biomarker. Complex networks of signaling pathways lead to increased cell proliferation, decreased cell adhesion, cellular differentiation, genetic instability, and other functions associated with the malignant phenotype. The purpose of this presentation is to illustrate the fundamental concepts for selection and profile analysis of high-level phenotypic biomarkers developed for bladder cancer risk assessment, screening, and early bladder cancer detection. PMID- 15358591 TI - The blood brain barrier in HIV infection. AB - The blood brain barrier (BBB) serves as a protective mechanism for the brain. It prevents entry of pernicious substances, whether chemical or cellular, from free access to the CNS. In essence, it is a defense mechanism preserving the internal milieu of the brain. The BBB may be disrupted by a number of pathological processes. CNS infection is a well recognized cause of BBB disruption. Among the CNS infections demonstrated to affect the BBB is human immunodeficiency virus (HIV). HIV infects the brain shortly after its acquisition. Studies of cerebrospinal fluid (CSF) as well as dynamic studies of contrast enhanced magnetic resonance imaging have confirmed abnormalities of the BBB in HIV infected persons. Pathological studies of the CNS have confirmed the in vivo studies, and in vitro studies have identified a range of pathogenic mechanisms of HIV-associated BBB compromise. This disruption of the BBB may not only contribute to accelerating brain infection by HIV, but may also alter CNS function. Additionally, BBB disruption has implications with respect to antiretroviral therapy. This review will address these issues. PMID- 15358592 TI - Tachykinin-mediated modulation of the immune response. AB - Tachykinins represent a family of peptides which have significant effects on such diverse physiological responses as gut peristalsis, vascular permeability, and the transmission of pain. Traditionally, neurons have been identified as the major source of tachykinin peptides in peripheral tissues and in the central nervous system, resulting in their classification as neuropeptides or, more specifically, as neurokinins. The fact that a variety of cell types can express one or more members of the family of neurokinin receptors accounts for the remarkable ability of these tachykinin peptides to affect multiple organ systems and numerous biological responses. Perhaps one of the most surprising findings during the last 15 years has been the understanding that expression of neurokinin receptors on leukocyte populations can be constitutive or inducible. This fact establishes a receptor-mediated mechanism by which tachykinin peptides can affect both innate and specific immune responses. Due to the numerous cytokines, chemokines, and other soluble mediators that can modulate leukocyte responses, the importance of tachykinin-mediated modulation of the immune response has met with some skepticism. The following reviews make a strong case for neurokinin receptor expression by leukocytes as a significant participant in host response. From hematopoiesis to antigen-specific T lymphocyte responses, tachykinins are present and can dramatically modulate maturation and responsiveness of these immune cells. Studies which document this surprising role for neurokinin receptor expression by leukocytes are detailed in the following reviews. PMID- 15358593 TI - Inhibition of mitochondrial gene transcription suppresses neurotensin secretion in the human carcinoid cell line BON. AB - Mitochondria, organelles essential for ATP production, play a central role in a number of cellular functions, including the regulation of insulin secretion. Neurotensin (NT), an important regulatory intestinal hormone, has been implicated in fatty acid translocation, gut motility and secretion, and intestinal cell growth; however, mechanisms regulating NT secretion have not been entirely defined. The purpose of this study was to determine the effect of inhibition of mitochondrial gene transcription on NT secretion. BON cells, a novel human carcinoid cell line that produces and secretes NT peptide and expresses the gene encoding NT (designated NT/N), were treated with ethidium bromide (EB; 0.05, 0.1, and 0.4 microg/ml), an inhibitor of DNA and RNA synthesis, or vehicle over a time course (1-4 days). Cells were then stimulated with either ACh (100 microM) or phorbol 12 myristate,13-acetate (PMA, 10 nM) for 30 min. Media and cells were extracted, and NT peptide measured by RIA. Treatment with EB had no effect on BON cell viability or cell cycle distribution over the 4-day course. In contrast, EB treatment produced a dose-dependent reduction of mitochondrial gene expression; however, NT/N gene expression was not altered. Mitochondrial inhibition by EB treatment suppressed NT secretion induced by ACh and PMA, both in a dose dependent manner. EB-mediated inhibition of NT secretion and mitochondrial gene expression was reversed with removal of EB. Our results demonstrate that inhibition of mitochondrial gene transcription suppresses both ACh- and PMA stimulated NT release. These findings are the first to demonstrate that mitochondrial function is important for agonist-mediated NT secretion. PMID- 15358594 TI - Differential subcellular targeting of PKC-epsilon in response to pharmacological or ischemic stimuli in intestinal epithelia. AB - Ischemia is the central pathogenic factor underlying a spectrum of intestinal disorders. The study of the cellular signaling responses to ischemic stress in nonepithelial cells has progressed substantially in the previous several years, but little is known about the response in epithelial cells. Unique features of the epithelial response to ischemic stress suggest differential regulation with regards to signaling. The PKC family of proteins has been implicated in ischemic stress in nonepithelial systems. The role of PKC isoforms in chemical ischemia in intestinal epithelial cells is evaluated in this study. Additionally, the phosphorylation of the F-actin cross-linking protein myristoylated alanine-rich C kinase substrate (MARCKS) is also studied. Chemical ischemia resulted in the transient activation of only the isoform PKC-epsilon as detected by translocation employing the subcellular fractionation technique. The pharmacological agonists phorbol 12-myristate 13-acetate and carbachol also led to the translocation of PKC-epsilon. By immunofluoresence, MARCKS is noted to be located at the lateral membrane under control conditions. In response to carbachol, MARCKS translocates to the cytosol, indicating its phosphorylation, which is additionally confirmed biochemically. Consistent with this observation, carbachol induces the translocation of PKC-epsilon to proximity with MARCKS at the lateral membrane. In response to chemical ischemia, MARCKS fails to translocate and phosphorylation does not increase. Additionally, the translocation of PKC-epsilon is not to the lateral membrane but rather basally. The data suggest that the differential translocation of PKC-epsilon in response to pharmacological agonists versus ischemic stress may lead to different effects on downstream targets. PMID- 15358595 TI - EGF receptor transactivation mediates ANG II-stimulated mitogenesis in intestinal epithelial cells through the PI3-kinase/Akt/mTOR/p70S6K1 signaling pathway. AB - The role of epidermal growth factor receptor (EGFR) tyrosine kinase and its downstream targets in the regulation of the transition from the G0/G1 phase into DNA synthesis in response to ANG II has not been previously investigated in intestinal epithelial IEC-18 cells. ANG II induced a rapid and striking EGFR tyrosine phosphorylation, which was prevented by selective inhibitors of EGFR tyrosine kinase activity (e.g., AG-1478) or by broad-spectrum matrix metalloproteinase (MMP) inhibitor GM-6001. Pretreatment of these cells with either AG-1478 or GM-6001 reduced ANG II-stimulated DNA synthesis by approximately 50%. To elucidate the downstream targets of EGFR, we demonstrated that ANG II stimulated phosphorylation of Akt at Ser473, mTOR at Ser2448, p70S6K1 at Thr389, and S6 ribosomal protein at Ser(235/236). Pretreatment with AG-1478 inhibited Akt, p70S6K1, and S6 ribosomal protein phosphorylation. Inhibition of phosphatidylinositol (PI)3-kinase with LY-294002 or mTOR/p70S6K1 with rapamycin reduced [3H]thymidine incorporation by 50%, i.e., to levels comparable to those achieved by addition of either AG-1478 or GM-6001. Utilizing Akt small interfering RNA targeted to Akt1 and Akt2, Akt protein knockdown dramatically inhibited p70S6K1 and S6 ribosomal protein phosphorylation. In contrast, AG-1478 or Akt gene silencing exerted no detectable inhibitory effect on ANG II-induced extracellular signal-regulated kinase 1/2 phosphorylation in IEC-18 cells. Taken together, our results demonstrate that EGFR transactivation mediates ANG II stimulated mitogenesis through the PI3-kinase/Akt/mTOR/p70S6K1 signaling pathway in IEC-18 cells. PMID- 15358596 TI - CXCL12 activation of CXCR4 regulates mucosal host defense through stimulation of epithelial cell migration and promotion of intestinal barrier integrity. AB - Intestinal epithelial cell migration plays a key role in gastrointestinal mucosal barrier formation, enterocyte development, differentiation, turnover, wound healing, and adenocarcinoma metastasis. Chemokines, through engagement of their corresponding receptors, are potent mediators of directed cell migration and are critical in the establishment and regulation of innate and adaptive immune responses. The aim of this study was to define the role for the chemokine CXCL12 and its sole cognate receptor CXCR4 in regulating intestinal epithelial cell migration and to determine its impact on barrier integrity. CXCL12 stimulated the dose-dependent chemotactic migration of human T84 colonic epithelial cells. Epithelial cell migration was inhibited by CXCR4 neutralizing antibody, pertussis toxin, LY-294002, and PD-98059, thereby implicating Galpha(i), phosphatidylinositol 3-kinase (PI3-kinase), and the ERK1/2 MAP kinase pathways in CXCR4-specific signaling. CXCL12 was also shown to increase barrier integrity, as defined by transepithelial resistance and paracellular flux across differentiating T84 monolayers. To determine whether CXCL12 regulated epithelial restitution, we used the normal nontransformed intestinal epithelial cell-6 (IEC 6) wound healing model. By using RT-PCR, immunoblot analysis, and immunofluorescence microscopy, we first showed expression of both CXCR4 and its ligand by IEC-6 cells. We then demonstrated that CXCL12 activated comparable signaling mechanisms to stimulate epithelial migration in the absence of proliferation in wounded IEC-6 monolayers. Taken together, these data indicate that CXCL12 signaling via CXCR4 directs intestinal epithelial cell migration, barrier maturation, and restitution, consistent with an important mechanistic role for these molecules in mucosal barrier integrity and innate host defense. PMID- 15358597 TI - Contribution of a time-dependent and hyperpolarization-activated chloride conductance to currents of resting and hypotonically shocked rat hepatocytes. AB - Hepatocellular Cl- flux is integral to maintaining cell volume and electroneutrality in the face of the many transport and metabolic activities that describe the multifaceted functions of these cells. Although a significant volume regulated Cl- current (VRAC) has been well described in hepatocytes, the Cl- channels underlying the large resting anion conductance have not been identified. We used a combination of electrophysiological and molecular approaches to describe potential candidates for this conductance. Anion currents in rat hepatocytes and WIF-B and HEK293T cells were measured under patch electrode voltage clamp. With K+-free salts of Cl- comprising the major ions externally and internally, hyperpolarizing steps between -40 and -140 mV activated a time dependent inward current in hepatocytes. Steady-state activation was half-maximal at -63 mV and 28-38% of maximum at -30 to -45 mV, previously reported hepatocellular resting potentials. Gating was dependent on cytosolic Cl-, shifting close to 58 mV/10-fold change in Cl- concentration. Time-dependent inward Cl- currents and a ClC-2-specific RT-PCR product were also observed in WIF B cells but not HEK293T cells. All cell types exhibited typical VRAC in response to dialysis with hypertonic solutions. DIDS (0.1 mM) inhibited the hepatocellular VRAC but not the inward time-dependent current. Antibodies against the COOH terminus of ClC-2 reacted with a protein between 90 and 100 kDa in liver plasma membranes. The results demonstrate that rat hepatocytes express a time-dependent inward Cl- channel that could provide a significant depolarizing influence in the hepatocyte. PMID- 15358598 TI - ADAM-15 inhibits wound healing in human intestinal epithelial cell monolayers. AB - The disintegrin metalloproteases (or ADAMs) are membrane-anchored glycoproteins that have been implicated in cell-cell or cell-matrix interactions and in proteolysis of molecules on the cell surface. The expression and/or the pathophysiological implications of ADAMs are not known in intestinal epithelial cells. Therefore, our aim was to investigate the expression and the role of ADAMs in intestinal epithelial cells. Expression of ADAMs was assessed by RT-PCR, Western blot analysis, and immunufluorescence experiments. Wound-healing experiments were performed by using the electric cell substrate impedence sensing technology. Our results showed that ADAMs-10, -12, and -15 mRNA are expressed in the colonic human cell lines Caco2-BBE and HT29-Cl.19A. An ADAM-15 complementary DNA cloned from Caco2-BBE poly(A)+ RNA, and encompassing the entire coding region, was found to be shorter and to present a different region encoding the cytoplasmic tail compared with ADAM-15 sequence deposited in the database. In Caco2-BBE cells and colonic epithelial cells, ADAM-15 protein was found in the apical, basolateral, and intracellular compartments. We also showed that the overexpression of ADAM-15 reduced cell migration in a wound-healing assay in Caco2-BBE monolayers. Our data show that 1) ADAM-15 is expressed in human intestinal epithelia, 2) a new variant of ADAM-15 is expressed in a human intestinal epithelial cell line, and 3) ADAM-15 is involved in intestinal epithelial cells wound-healing processes. Together, these results suggest that ADAM-15 may have important pathophysiological roles in intestinal cells. PMID- 15358599 TI - sPAR-3, a splicing variant of PAR-3, shows cellular localization and an expression pattern different from that of PAR-3 during enterocyte polarization. AB - PAR-3 (partitioning-defective) is a scaffold-like PDZ (postsynaptic density 95/discs large/zonula occludens-1) domain-containing protein that forms a complex with PAR-6 and atypical PKC, localizes to tight junctions, and contributes to the formation of functional tight junctions. There are several alternatively spliced isoforms of PAR-3, although their physiological significance remains unknown. In this study, we show that one of the major isoforms of PAR-3, sPAR-3, is predominantly expressed in the Caco-2 cells derived from colon carcinoma and is used as a model to investigate the events involved in the epithelial cell differentiation and cell polarity development. During the polarization of Caco-2 cells, the expression of PAR-3 increases as do those of other cell-cell junction proteins, whereas the expression of sPAR-3 decreases. Biochemical characterization revealed that sPAR-3 associates with atypical PKC, as does PAR 3. On the other hand, immunofluorescence microscopy revealed that sPAR-3 does not concentrate at the cell-cell contact region in fully polarized cells, whereas it concentrates at premature cell-cell junctions. This makes a contrast to PAR-3, which concentrates at tight junctions in fully polarized cells. These results provide evidence suggesting the difference in the role between sPAR-3 and PAR-3 in epithelial cells. PMID- 15358600 TI - Common promoter C516T polymorphism in the ApoB gene is an independent predictor of carotid atherosclerotic disease in subjects presenting a broad range of plasma cholesterol levels. AB - OBJECTIVE: A common polymorphism in the promoter of the apolipoprotein B (apoB) gene, a C to T change at position -516, increases the transcription rate of apoB, resulting in elevated circulating levels of low-density lipoprotein (LDL) cholesterol. METHODS AND RESULTS: We tested the hypothesis that carriers of the 516T allele, who may display consistent elevation in plasma cholesterol over their lifetime, may present more extensive atherosclerotic disease than noncarriers. Genotyping of the apoB 516 C/T promoter polymorphism was performed in 326 subjects at low cardiovascular risk. Homozygotes for allele T displayed higher plasma levels of apoB and LDL than did heterozygotes. Furthermore, both homozygotes and heterozygotes for allele T exhibited higher plasma levels of apoB and LDL than did homozygotes for allele C (P<0.0001). In addition, homozygotes for allele T displayed higher carotid intima-media thickness (IMT) than subjects who were heterozygous. Moreover, both groups had higher carotid IMT than subjects of genotype -516C/C (P<0.001). Only age, high-density lipoprotein, and the presence of allele T were identified as independent predictors of the presence of carotid plaque. No association existed between the polymorphism and plasma concentrations of triglycerides, high-density lipoprotein, or apoAI. CONCLUSIONS: Our data indicate that a C to T change at position -516 of the apoB gene is independently associated with the presence of carotid atherosclerotic disease. Identification of the -516C/T polymorphism may therefore contribute to the estimation of overall cardiovascular risk. PMID- 15358601 TI - Cyclosporin A traps ABCA1 at the plasma membrane and inhibits ABCA1-mediated lipid efflux to apolipoprotein A-I. AB - OBJECTIVE: ABCA1 mediates cellular cholesterol and phospholipid efflux to apolipoprotein A-I and other apolipoprotein acceptors. In this study, we analyzed the effect of the immunosuppressant cyclosporin A on the ABCA1-mediated lipid effluxes reactions. METHODS AND RESULTS: Cyclosporin A acted as a potent inhibitor of ABCA1 activity in several cell lines. Using the RAW264.7 mouse macrophage cell line, in which ABCA1 and its associated cholesterol efflux activity are inducible by cAMP analogues, cyclosporin A inhibition of cholesterol efflux to apolipoprotein A-I was rapidly reversible after its removal from the culture media, implying that ABCA1 levels were not drastically reduced by cyclosporin A. In fact, cyclosporin A treatment decreased ABCA1 turnover and yielded a 2-fold increase in cell-surface ABCA1. Despite the increase in cell surface ABCA1, cyclosporin A decreased apolipoprotein A-I uptake, resecretion, and degradation in RAW cells. Finally, consistent with the inhibition of ABCA1 in vitro, cyclosporin A treatment induced a 33% reduction of high-density lipoprotein (HDL) levels in mice. CONCLUSIONS: ABCA1 inhibition by cyclosporin A supports a role for ABCA1 endocytic trafficking in ABCA1-mediated lipid efflux and could explain in part the low HDL levels observed in some patients with transplants. PMID- 15358602 TI - Sonic hedgehog induces arteriogenesis in diabetic vasa nervorum and restores function in diabetic neuropathy. AB - OBJECTIVE: The embryonic morphogen sonic hedgehog (SHh) has been shown to induce neovascularization of ischemic tissue but has not been shown to play a role in regulating vascular nerve supply. Accordingly, we investigated the hypothesis that systemic injection of SHh protein could improve nerve blood flow and function in diabetic neuropathy (DN). METHODS AND RESULTS: Twelve weeks after induction of diabetes with streptozotocin, motor and sensory nerve conduction velocities (MCV and SCV) of the sciatic nerves were significantly reduced in diabetic rats. SHh-treated diabetic rats demonstrated marked improvement of both MCV and SCV (P<0.05). Laser Doppler perfusion imaging showed that nerve blood flow was significantly reduced in the diabetic rats but was restored in SHh treated diabetic rats (P<0.05 versus diabetic saline-treated rats) to levels similar to those achieved with vascular endothelial growth factor-2 (VEGF-2) gene therapy. In vivo perfusion of Bandeuraea simplicifolia (BS)-1 lectin showed marked reduction in the vasa nervora in diabetic sciatic nerves but restoration of nerve vasculature to nondiabetic levels in the SHh-treated and plasmid DNA encoding human VEGF-2 (phVEGF-2)-treated diabetic nerves. Interestingly, the SHh induced vasculature was characterized by larger diameter and more smooth muscle cell-containing vessels, compared with VEGF-2 gene-treated diabetic rats. CONCLUSIONS: These data indicate that Shh induces arteriogenesis and restores nerve function in DN. PMID- 15358603 TI - Hypoxia increases LDL oxidation and expression of 15-lipoxygenase-2 in human macrophages. AB - OBJECTIVE: Macrophage-mediated oxidation of low-density lipoprotein (LDL) by enzymes, such as the lipoxygenases, is considered of major importance for the formation of oxidized LDL during atherogenesis. Macrophages have been identified in hypoxic areas in atherosclerotic plaques. METHODS AND RESULTS: To investigate the role of hypoxia in macrophage-mediated LDL oxidation, we incubated human monocyte-derived macrophages with LDL under normoxic (21% O2) or hypoxic (0% O2) conditions. The results showed that hypoxic macrophages oxidized LDL to a significantly higher extent than normoxic cells. Interestingly, the mRNA and protein expression of 15-lipoxygenase-2 (15-LOX-2) as well as the activity of this enzyme are elevated in macrophages incubated at hypoxia. Both the unspliced 15-LOX-2 and the spliced variant 15-LOX-2sv-a are found in macrophages. In addition, 15-LOX-2 was identified in carotid plaques in some macrophage-rich areas but was only expressed at low levels in nondiseased arteries. CONCLUSIONS: In summary, these observations show for the first time that 15-LOX-2 is expressed in hypoxic macrophages and in atherosclerotic plaques and suggest that 15-LOX-2 may be one of the factors involved in macrophage-mediated LDL oxidation at hypoxia. PMID- 15358604 TI - Maturational modulation of endothelium-dependent vasodilatation in ovine cerebral arteries. AB - To address the hypothesis that maturation enhances endothelial vasodilator function in cerebral arteries, relaxant responses to ADP and A-23187 were determined in ovine carotid and cerebral arteries harvested from 25 newborn lambs (3-7 days) and 23 adult sheep. Maturation significantly increased pD(2) values for A-23187 (newborn range: 4.9 +/- 0.3 to 5.4 +/- 0.3; adult range: 6.0 +/- 0.2 to 7.1 +/- 0.2) and the maximal vasodilator response to A-23187 by 10-18%. In contrast, maturation decreased maximum responses to ADP by 5-25% with no change in pD(2). The magnitudes of endothelium-dependent relaxation were not affected by 10 microM indomethacin but were virtually abolished by 100 microM N(G)-nitro-L arginine methyl ester/L-nitro arginine, indicating that nitric oxide (NO) is the primary endothelium-dependent vasodilator in these arteries. Maturation also modestly decreased endothelial NO synthase (eNOS) abundance in both carotid (32%) and cerebral (26%) arteries. Together, these findings reinforce the view that receptor coupling to endothelial activation is tightly regulated and may offset underlying changes in maximal endothelial vasodilator capacity. This capacity, in turn, appears to increase with postnatal age despite major growth and expansion of endothelial cell size and vascular wall volume. In ovine cerebral arteries, endothelial vasodilator capacity appears completely dependent on eNOS activity but not on cyclooxygenase activity. In turn, eNOS activity appears to be postnatally regulated by mechanisms independent of changes in eNOS abundance alone. PMID- 15358605 TI - Intracerebroventricular administration of MK-801 increases food intake through mechanisms independent of gastric emptying. AB - Systemic or hindbrain administration of MK-801, a noncompetitive N-methyl-D aspartate receptor antagonist, increases meal size. To examine whether MK-801 enhances intake by increasing gastric emptying, we administered MK-801 (2.0 microg/3.0 microl) into the fourth ventricle [intracerebroventricular (ICV)] and measured feeding and gastric emptying of 5-ml NaCl or 15% sucrose loads. In a parallel experiment, we examined food intake and gastric emptying following intraperitoneal (IP) injection of MK-801 (100 microg/kg). MK-801, either IP or ICV, increased 30-min sucrose intake compared with control (12.3 +/- 0.7 vs. 9.8 +/- 0.5 and 16.6 +/- 2.0 vs. 10.7 +/- 0.7 ml, for IP and ICV administration, respectively). Also, IP MK-801 increased 5-min gastric emptying of NaCl (4.13 +/- 0.1 ml emptied) and sucrose (3.11 +/- 0.1 ml emptied) compared with control (3.75 +/- 0.2 and 2.28 +/- 0.1 ml emptied for NaCl and sucrose loads, respectively). In contrast, ICV MK-801 did not alter NaCl emptying (3.82 +/- 0.1 ml emptied) compared with control (3.82 +/- 0.3 ml emptied) and actually reduced gastric emptying of sucrose (2.1 +/- 0.2 and 2.94 +/- 0.1 ml emptied, for MK and vehicle, respectively). These data confirm previous results that systemic as well as hindbrain injection of MK-801 increases food intake. However, because ICV MK-801 failed to increase gastric emptying, these results indicate that MK-801 increases food intake through mechanisms independent of altered gastric emptying. PMID- 15358606 TI - Increased oral and decreased intestinal sensitivity to sucrose in obese, prediabetic CCK-A receptor-deficient OLETF rats. AB - CCK-A receptor-deficient Otsuka Long-Evans Tokushima fatty (OLETF) rats are hyperphagic and develop obesity and Type 2 diabetes. In this strain, taste preference functions have not been investigated. Therefore, a series of short access, two-bottle tests were performed in age-matched prediabetic OLETF and nonmutant Long-Evans Tokushima Otsuka (LETO) rats to investigate preference for sucrose (0.03, 0.1, 0.3, or 1.0 M) presented with a choice of water. To discern orosensory from postgastric factors that may contribute to this preference, in a separate experiment, rats were allowed to sham feed sucrose in the absence or presence of duodenal sucrose infusion (0.3, 0.6, or 1.0 M). In the two-bottle real-feeding tests, OLETF rats exhibited a greater preference for 0.3 M sucrose (91.2 +/- 1.7 and 78.5 +/- 3.4% for OLETF and LETO, respectively; P < 0.01) and 1.0 M sucrose (65.3 +/- 1.2 and 57.5 +/- 2.7% for OLETF and LETO, respectively; P < 0.05) than LETO rats. OLETF rats also sham fed less of the lowest (0.03 M; 33.8 +/- 4.8 and 58.3 +/- 7.3 ml for OLETF and LETO, respectively; P < 0.05) and more of the highest (1.0 M; 109.9 +/- 6.5 and 81.0 +/- 3.9 ml for OLETF and LETO, respectively; P < 0.01) concentration of sucrose relative to LETO rats. Finally, intraduodenal sucrose infusions (0.6 and 1.0 M) produced a smaller reduction of 0.3 M sham sucrose intake [14.1 +/- 8.1 vs. 52.5 +/- 3.3 ml and 49.4 +/- 8.0 vs. 82.4 +/- 3.2 ml for 0.6 M (P < 0.01) and 1.0 M (P < 0.05) infusions in OLETF and LETO, respectively]. These findings demonstrate that OLETF rats display an increased preference for sucrose, an effect that is at least partially influenced by the orosensory stimulating effect of sucrose. This enhanced responsiveness to oral stimulation, coupled with the deficit in responding to the postingestive feedback of intestinal sucrose, may contribute additively to the development of hyperphagia and weight gain in OLETF rats. PMID- 15358607 TI - Biphasic temporal pattern in exercise capacity after myocardial infarction in the rat: relationship to left ventricular remodeling. AB - After myocardial infarction (MI), there is progressive left ventricular (LV) remodeling and impaired exercise capacity. We tested the hypothesis that LV remodeling results in structural and functional changes that determine exercise impairment post-MI. Rats underwent coronary artery ligation (n = 12) or sham (n = 11) surgery followed by serial exercise tests and echocardiography for 16 wk post MI. LV pressure-volume relationships were determined using a blood-perfused Langendorff preparation. Exercise capacity was 60% of shams immediately post-MI (P < 0.05) followed by a recovery to near normal during weeks 5-8. Thereafter, there was a progressive decline in exercise capacity to +/-40% of shams (P < 0.01). At both 8 and 16 wk post-MI, fractional shortening (FS) was reduced and end-diastolic diameter (EDD) was increased (P < 0.01). However, neither FS nor EDD correlated with exercise at 8 or 16 wk (r(2) < 0.12, P > 0.30). LV septal wall thickness was increased at both 8 (P = 0.17 vs. shams) and 16 wk (P = 0.035 vs. shams) post-MI and correlated with exercise at both times (r(2) >/= 0.50 and P T at Cys43 and 3462C>T at Leu120, and 2 missense mutations, Asp38Tyr and Arg151Gln, were identified. None of these mutations was found to affect the interindividual variability of warfarin prescribed. Finally, 2 common polymorphisms were found, 1173C>T in the intron 1 and 3730G>A transition in the 3' untranslated region (UTR). Regardless of the presence of confounding variables, the mean adjusted dose required of warfarin was higher (6.2 mg) among patients with the VKORC1 1173CC genotype than those of patients carrying the CT (4.8 mg; P = .002) or the TT genotype (3.5 mg; P < .001). In the present setting, VKORC1 and CYP2C9 genetic variants investigated accounted for about a third (r2, 0.353) of the interindividual variability. Genetic variants of the VKORC1 gene locus modulate the mean daily dose of drug prescribed to acquire the target anticoagulation intensity. PMID- 15358624 TI - Hmgb3 deficiency deregulates proliferation and differentiation of common lymphoid and myeloid progenitors. AB - Hmgb3 is an X-linked member of a family of chromatin-binding proteins that is expressed in primitive hematopoietic cells capable of long-term hematopoietic repopulation. To examine the role of Hmgb3 in adult hematopoiesis, we generated Hmgb3-deficient (Hmgb3(-/Y)) mice, which are viable but erythrocythemic. Hmgb3( /Y) mice contain normal numbers of hematopoietic stem cells (HSCs), which generate fewer than normal numbers of common lymphoid progenitors (CLPs) and common myeloid progenitors (CMPs) and greater than normal numbers of more mature progenitors. Although fewer Hmgb3(-/Y) primitive progenitor cells are in the G2/M cell cycle phase, bromodeoxyuridine (BrdU) incorporation demonstrated enhanced proliferation compared with their wild-type counterparts. Hmgb3(-/Y) HSCs have increased levels of Gata-2 and c-myb mRNA. We propose that Hmgb3 deficiency leads to a failure of HSCs to expand into normal numbers of CLPs and CMPs. This defect is compensated for by the ability of Hmgb3(-/Y) progenitors to expand rapidly and differentiate into normal numbers of hematopoietic cells. PMID- 15358625 TI - Proinflammatory mediators elicit secretion of the intracellular B-lymphocyte stimulator pool (BLyS) that is stored in activated neutrophils: implications for inflammatory diseases. AB - We have recently shown that granulocyte-colony-stimulating factor (G-CSF)- and interferon-gamma (IFN-gamma)-activated human neutrophils accumulate and release remarkable amounts of soluble B-lymphocyte stimulator (BLyS) in vitro. In this study, we provide evidence that neutrophils migrating into skin window exudates (SWEs) developed in healthy volunteers and in patients with rheumatoid arthritis (RA), synthesized, and released BLyS in response to locally produced G-CSF. Accordingly, the concentrations of soluble BLyS in SWEs were significantly more elevated than in serum. Because the levels of SWE BLyS, but not SWE G-CSF, were higher in patients with RA than in healthy subjects, we examined the effect of CXCL8/IL-8, C5a, and other proinflammatory mediators that dramatically accumulate in RA SWEs and in inflamed synovial fluids. We show that CXCL1/GROalpha, CXCL8/IL 8, C5a, immune complexes, tumor necrosis factor-alpha (TNF-alpha), leukotriene B4, N-formyl-methionyl-leucyl-phenylalanine (fMLP), and lipopolysaccharide (LPS), which by themselves do not induce BLyS de novo synthesis, act as potent secretagogues for BLyS, which is mainly stored in Golgi-related compartments within G-CSF-treated neutrophils, as determined by immunogold electron microscopy. This action is pivotal in greatly amplifying neutrophil-dependent BLyS release in SWEs of patients with RA compared with healthy subjects. Collectively, our data uncover a novel mechanism that might dramatically exacerbate the release of BLyS by neutrophils during pathologic inflammatory responses. PMID- 15358626 TI - Acquired alpha-thalassemia in association with myelodysplastic syndrome and other hematologic malignancies. AB - Abnormalities of hemoglobin synthesis are usually inherited but may also arise as a secondary manifestation of another disease, most commonly hematologic neoplasia. Acquired hemoglobin disorders can be seen in any population and are not restricted to areas of the world with high incidences of inherited hemoglobinopathies. In fact, the acquired hemoglobinopathies may be more readily recognized where inherited hemoglobin abnormalities are rare and less likely to cause diagnostic confusion. Acquired alpha-thalassemia is the best characterized of the acquired red blood cell disorders in patients with hematologic malignancy, and it is almost always associated with a myelodysplastic syndrome (MDS). At least 2 molecular mechanisms for acquired alpha-thalassemia are now recognized: acquired deletion of the alpha-globin gene cluster limited to the neoplastic clone and, more commonly, inactivating somatic mutations of the trans-acting chromatin-associated factor ATRX, which cause dramatic down-regulation of alpha globin gene expression. Here we review the clinical, hematologic, and molecular genetic features of alpha-thalassemia arising in a clonal myeloid disorder, and we discuss howATRX might affect gene expression in normal and abnormal hematopoiesis through epigenetic mechanisms. PMID- 15358627 TI - Murine embryonic stem cell differentiation is promoted by SOCS-3 and inhibited by the zinc finger transcription factor Klf4. AB - Embryonic stem (ES) cells homozygous for a Shp-2 mutation (Shp-2(Delta46-110)) demonstrate leukemia inhibitory factor (LIF) hypersensitivity and increased LIF stimulated phosphorylation of signal transducer and activator of transcription (STAT3). We hypothesized that LIF-responsive genes in Shp-2(Delta46-110) cells would represent potential candidates for molecules vital for ES cell self renewal. Using microarray analysis, we detected 41 genes whose expression was modified by LIF in Shp-2(Delta46-110) ES cells. Induction of 2 significantly up regulated genes, suppressor of cytokine signaling-3 (SOCS-3) and Kruppel-like factor 4 (Klf4), was verified using Northern blotting. ES cells overexpressing SOCS-3 had an increased capacity to differentiate to hematopoietic progenitors, rather than to self-renew. In contrast, ES cells overexpressing Klf4 had a greater capacity to self-renew based on secondary embryoid body (EB) formation. Klf4-transduced d6 EBs expressed higher levels of Oct-4, consistent with the notion that Klf4 promotes ES cell self-renewal. These findings verify the negative role of SOCS-3 on LIF signaling and provide a novel role for Klf4 in ES cell function. PMID- 15358628 TI - Vascular leukocytes contribute to tumor vascularization. AB - There is no proof that hematopoietic cells contribute significantly to vasculogenesis in postnatal life. Here we report a novel leukocyte subset within ovarian carcinoma that coexpresses endothelial and dendritic cell markers. Fluorescence-activated cell sorter (FACS) analysis identified a high frequency of VE-cadherin+ CD45+ leukocytes (39% of host cells) in 10 of 10 solid tumors evaluated. This population represented less than 1% of nontumor cells in ascites and peripheral blood. At the protein level, more than 86% of these cells expressed the endothelial markers P1H12, CD34, and CD31 and leukocyte markers CD11c and major histocompatibility complex (MHC) class II. At the mRNA level, we detected TEM1, TEM7, and Thy-1, specific markers of angiogenic endothelium. Finally, this population has the capacity to generate functional blood vessels in vivo. Because of its mixed phenotype, we named this population vascular leukocytes (VLCs). Our data provide an important link between hematopoietic endothelial precursors and vascular development in postnatal life and a possible novel therapeutic target. PMID- 15358629 TI - High incidence of occult leptomeningeal disease detected by flow cytometry in newly diagnosed aggressive B-cell lymphomas at risk for central nervous system involvement: the role of flow cytometry versus cytology. AB - We assessed the cerebrospinal fluid (CSF) by flow cytometry and cytology in 51 newly diagnosed and 9 treated aggressive B-cell lymphomas at risk for central nervous system (CNS) involvement to examine the utility of flow cytometry, incidence of CSF disease, and clinical surrogates of CNS spread. Multicolor flow cytometry using multiple antibody panels for light chains and B- and T-cell antigens identified neoplastic clones that constituted as little as 0.2% of total CSF lymphocytes. Among 51 newly diagnosed patients, 11 (22%) had occult CSF involvement. All 11 were detected by flow cytometry but only 1 by cytology (P = .002). Among 9 treated patients, CSF involvement was detected by flow cytometry alone in 2 and also by cytology in 1 case. CSF chemistry and cell counts were similar in patients with and without CSF lymphoma. Only the number of extranodal sites was associated with occult CSF lymphoma in newly diagnosed patients by univariate (P = .006) or logistic regression analysis (P = .012). We hypothesize that the biologic phenotype associated with colonization of extranodal sites leads to CNS spread, possibly related to the microenvironment. Patients at risk for CNS spread should undergo staging CSF evaluation by flow cytometry. PMID- 15358630 TI - Prediction of T-cell reconstitution by assessment of T-cell receptor excision circle before allogeneic hematopoietic stem cell transplantation in pediatric patients. AB - The extent and rapidity with which T cells are regenerated from graft-derived precursor cells directly influences the incidence of infection and the T-cell based graft-versus-tumor effect. Measurement of T-cell receptor excision circles (TRECs) in peripheral blood is a means of quantifying recent thymic T-cell production and has been used after transplantation in many studies to estimate thymus-dependent T-cell reconstitution. We hypothesized that the quality of thymic function before transplantation affects thymus-dependent T-cell reconstitution after transplantation. We used real-time polymerase chain reaction (PCR) to quantify signal-joint TRECs (sjTRECs) before and after transplantation. T-cell reconstitution was evaluated by T-cell receptor beta (TCRbeta) CDR3 size spectratyping. We tested 77 healthy sibling donors and 244 samples from 26 pediatric recipients of allogeneic hematopoietic stem cell transplantation (AHSCT). Blood from the healthy donors contained 1200 to 155,000 sjTREC copies/mL blood. Patients who had greater than 1200 copies/mL blood before transplantation showed early recovery of sjTREC numbers and TCRbeta repertoire diversity. In contrast, patients who had fewer than 1200 copies/mL blood before transplantation demonstrated significantly slower restoration of thymus-dependent T cells. We conclude that the rate of reconstitution of thymus-dependent T cells is dependent on the competence of thymic function in the recipients before transplantation. Therefore, pretransplantation measurement of sjTREC may provide an important tool for predicting thymus-dependent T-cell reconstitution after transplantation. PMID- 15358631 TI - The chemopreventive action of catechins in the TRAMP mouse model of prostate carcinogenesis is accompanied by clusterin over-expression. AB - Clusterin (CLU) protein is widely distributed in animal tissues and is involved in many different processes, including apoptosis and neoplastic transformation. Green tea catechins (GTC) are known to exert chemopreventive effects in many cancer models, including transgenic adenocarcinoma mouse prostate (TRAMP) mice that spontaneously develop prostate cancer (CaP). We report here that growth of SV40-immortalized human prostate epithelial cells (PNT1A) as well as tumorigenic, poorly differentiated prostate cancer cells (PC-3) was potently inhibited by EGCG, the major green tea catechin, while normal human prostate epithelial cells were not significantly affected. IC(50) doses of EGCG for 24 h caused caspase cascade activation and CLU protein accumulation in both cells lines but not in normal cells, in which CLU remained undetectable. While 100% of TRAMP mice developed CaP, only 20% of those receiving 0.3% GTC in drinking water developed the neoplasm. In TRAMP mice, the CLU gene was dramatically down-regulated during onset and progression of CaP. In GTC-treated TRAMP mice in which tumor progression was chemoprevented, CLU mRNA and protein progressively accumulated in the prostate gland. CLU dropped again to undetectable levels in animals in which GTC chemoprevention failed and CaP developed. Up-regulation of histone H3 and down-regulation of growth arrest-specific gene 1 (Gas1) mRNAs in CaP-developing TRAMP mice demonstrated a high proliferation rate in tumors, while the opposite occurred in the glands of GTC chemoprevented animals. Failure of GTC chemoprevention caused induction of both histone H3 and Gas1 and down-regulation of CLU. Immunohistochemistry experiments confirmed CLU down-regulation during CaP onset and progression, and CLU sustained expression in chemoprevented TRAMP mice. A possible role for CLU as a novel tumor-suppressor gene in the prostate is thus suggested. PMID- 15358632 TI - Modeling metastasis in vivo. AB - Metastasis, the spread of a tumor from its primary site to other parts of the body, continues to be the most significant problem in the field of cancer. Patients who present with metastatic disease or those who develop metastases after successful management of the primary tumor carry a universally grave prognosis. To improve treatment outcomes for these patients a broader understanding of the biology of metastases is necessary. The biological complexity that characterizes metastasis requires complex experimental systems for its study. To a large extent the modeling of this biological complexity is only possible using animal models. The following review will summarize the strengths and weaknesses of available in vivo models of metastasis including transplantable syngeneic mouse and human-mouse xenografts, genetically engineered mice and naturally occurring cancers of companion animals (pet dogs and cats). No single metastasis model is sufficient to answer all questions. As such, the selection of the optimal model(s) for each biological or translational question is necessary. PMID- 15358633 TI - n-3 PUFAs reduce VEGF expression in human colon cancer cells modulating the COX 2/PGE2 induced ERK-1 and -2 and HIF-1alpha induction pathway. AB - n-3 Polyunsaturated fatty acids (PUFAs) inhibit the development of microvessels in mammary tumors growing in mice. Human colorectal tumors produce vascular endothelial growth factor (VEGF) whose expression is up-regulated in tumor cells by both cyclooxygenase-2 (COX-2) and PGE(2) and directly correlated to neoangiogenesis and clinical outcome. The goal of this study was to examine the capability of n-3 PUFAs to regulate VEGF expression in HT-29 human colorectal cells in vitro and in vivo. Constitutive VEGF expression was augmented in cultured HT-29 cells by serum starvation and the effects of eicosapentaenoic (EPA) or docosahexaenoic acid (DHA) on VEGF, COX-2, phosphorylated extracellular signal-regulated kinase (ERK)-1 and -2 and hypoxia-inducible-factor 1-alpha (HIF 1alpha) expression and PGE(2) levels were assessed. Tumor growth, VEGF, COX and PGE(2) analysis were carried out in tumors derived from HT-29 cells transplanted in nude mice fed with either EPA or DHA. Both EPA and DHA reduced VEGF and COX-2 expression and PGE(2) levels in HT-29 cells cultured in vitro. Moreover, they inhibited ERK-1 and -2 phosphorylation and HIF-1alpha protein over-expression, critical steps in the PGE(2)-induced signaling pathway leading to the augmented expression of VEGF in colon cancer cells. EPA always showed higher efficacy than DHA in vitro. Both fatty acids decreased the growth of the tumors obtained by inoculating HT-29 cells in nude mice, microvessel formation and the levels of VEGF, COX-2 and PGE(2) in tumors. The data provide evidence that these n-3 PUFAs are able to inhibit VEGF expression in colon cancer cells and suggest that one possible mechanism involved may be the negative regulation of the COX-2/PGE(2) pathway. Their potential clinical application as anti-angiogenic compounds in colon cancer therapy is proposed. PMID- 15358634 TI - Fatty acid synthase is a potential molecular target for the chemoprevention of breast cancer. AB - The purpose of this investigation was to determine whether fatty acid synthase (FAS) is a potential molecular target for the chemoprevention of breast cancer by evaluating the effect of the FAS inhibitor triclosan on rat mammary carcinogenesis. At 50 days of age, 60 female Sprague-Dawley rats received 50 mg/kg methylnitrosourea (MNU) i.p. to initiate mammary carcinogenesis. One week later, half of the rats were fed triclosan at a level of 1000 p.p.m. in an AIN 93G diet for the remainder of the experiment. The other 30 control rats were fed an AIN-93G diet without triclosan. Twelve weeks after MNU treatment, 70% of control rats had mammary adenocarcinomas compared with only 43.3% of the triclosan group (P < 0.05). The control rats had an average of 2.7 +/- 0.3 tumors/rat compared with 1.8 +/- 0.3 in the triclosan group (P < 0.05). Western analysis showed that the tumors in the control rats expressed high levels of FAS. Immunohistochemistry showed that sections of tumors that stained strongly for FAS also showed strong staining for proliferating cell nuclear antigen. Furthermore, at biologically relevant dose levels, triclosan inhibited the activity of FAS in mammary tumor homogenates. These results indicate that triclosan suppresses rat mammary carcinogenesis by inhibiting FAS and suggest that FAS is a promising molecular target for breast cancer chemoprevention. PMID- 15358635 TI - Expression of macrophage migration inhibitory factor in esophageal squamous cell carcinoma and effects of bile acids and NSAIDs. AB - The aim of this study was to determine the macrophage migration inhibitory factor (MIF) protein and mRNA expression in esophageal squamous cell carcinoma (ESCC), and the effect of bile acids, aspirin and a selective cyclooxygenase-2 (COX-2) inhibitor, NS398, on MIF expression in ESCC cells in vitro. Specimens from tumors and the adjacent non-cancerous tissues were obtained from 52 ESCC patients. Western blotting was used for the detection of MIF protein expression, and reverse transcription-polymerase chain reaction (RT-PCR) for MIF mRNA expression. Cells of an ESCC cell line, Eca-109, were treated with chenodeoxycholate (CD, 100 mM), glycochenodeoxycholate (GCD, 1 mM), aspirin (1 mM) or NS398 (1 microM). Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were used to detect the expression of the MIF protein and mRNA, respectively, in the supernatant and cultured cells. Western blotting demonstrated that levels of MIF protein were increased in tumors versus non-malignant tissues, with the expression ratio of MIF over beta-actin of 0.93 +/- 0.21 and 0.57 +/- 0.08, respectively (P = 0.012). In vitro, both CD and GCD induced a dramatic increase in MIF protein and mRNA in ESCC cells. On the other hand, aspirin and NS398 significantly decreased MIF protein and mRNA expression, and completely blocked bile acid-induced MIF synthesis in the presence or absence of prostaglandin E(2). In conclusion, MIF expression is increased in ESCC. Whereas bile acids induce MIF expression in ESCC cells, aspirin and NS398 significantly inhibit MIF expression, even in the presence of bile acids, via a COX-independent mechanism. PMID- 15358636 TI - NAD+-linked 15-hydroxyprostaglandin dehydrogenase (15-PGDH) behaves as a tumor suppressor in lung cancer. AB - It has been reported that two inducible prostaglandin synthetic enzymes, cylooxygenase-2 (COX-2) and microsomal PGE synthase, are over-expressed in non small cell lung cancer (NSCLC). Using quantitative reverse transcription polymerase chain reaction, we analyzed RNA levels of the key prostaglandin catabolic enzyme, NAD+-linked 15-hydroxyprostaglandin dehydrogenase (15-PGDH), in 19 pairs of NSCLC tumors and adjacent non-malignant tissue from the same patient. We found that 100% of tumor-tissue pairs showed at least a 2-fold decrease and 61% showed a 10-fold decrease. This suggests that the increased expression of COX 2 and PGE synthase in tumors may work in concert with the decreased expression of 15-PGDH to amplify an increase in tissue levels of proliferative PGE2. To further explore if 15-PGDH is related to tumorigenesis, athymic nude mice were injected with control A549 cells or cells transiently over-expressing wild-type or mutant 15-PGDH (Y151F). It was found that mice injected with control A549 cells or with cells expressing mutant enzyme produced tumors normally. However, mice injected with A549 cells expressing wild-type 15-PGDH had a significant decrease in tumor growth. Examining the effects of 15-PGDH expression on cellular changes in A549 cells, we found that over-expression of 15-PGDH induced apoptosis of A549 cells as evidenced by fragmentation of DNA, activation of pro-caspase 3, cleavage of poly(ADP-ribose) polymerase and decreased expression of Bcl-2. We also found that the expression of 15-PGDH was negatively related to that of pro-adhesive and invasive CD44. Furthermore, the expression of 15-PGDH was found to be stimulated by hyaluronidase. These results suggest that 15-PGDH may decrease the level of proliferative PGE2, induce apoptosis and function like a tumor suppressor. PMID- 15358637 TI - Mitochondrial disorders. AB - In the medical literature the term 'mitochondrial disorders' is to a large extent applied to the clinical syndromes associated with abnormalities of the common final pathway of mitochondrial energy metabolism, i.e. oxidative phosphorylation (OXPHOS). Faulty oxidative phosphorylation may be due to overall dysfunction of the respiratory chain, a heteromultimeric structure embedded in the inner mitochondrial membrane, or can be associated with single or multiple defects of the five complexes forming the respiratory chain itself. From the genetic standpoint, the respiratory chain is a unique structure of the inner mitochondrial membrane formed by means of the complementation of two separate genetic systems: the nuclear genome and the mitochondrial genome. The nuclear genome encodes the large majority of the protein subunits of the respiratory complexes and most of the mitochondrial DNA (mtDNA) replication and expression systems, whereas the mitochondrial genome encodes only 13 respiratory complex subunits, and some RNA components of the mitochondrial translational apparatus. Accordingly, mitochondrial disorders due to defects in OXPHOS include both mendelian-inherited and cytoplasmic-inherited diseases. This review describes human genetic diseases associated with mtDNA and nuclear DNA mutations leading to impaired OXPHOS. PMID- 15358638 TI - Interleukin 8 secretion from monocytes of subjects heterozygous for the deltaF508 cystic fibrosis transmembrane conductance regulator gene mutation is altered. AB - Patients with cystic fibrosis (CF) exhibit an excessive host inflammatory response. The aim of this study was to determine (i) whether interleukin 8 (IL-8) secretion is increased from monocytes from subjects heterozygous as well as homozygous for cystic fibrosis transmembrane conductance regulator (CFTR) mutations and (ii) whether this is due to increased cell surface lipopolysaccharide (LPS) receptors or, alternatively, increased activation of mitogen-activated protein kinases (MAPK). The basal level of IL-8 secretion was higher from monocytes from CF patients than from monocytes from healthy controls (P = 0.02) and obligate heterozygotes (parents of the CF patients). The 50% effective concentrations for LPS-induced IL-8 production for monocytes from both CF patients and obligate heterozygotes were 100-fold lower than those for monocytes from healthy controls (P < 0.05). No differences in the levels of IL 1beta production were seen between these groups. Expression of the LPS surface receptors CD14 and Toll-like receptor 4 were not different between CF patients and healthy controls. In contrast, phosphorylation of the MAPKs p38 and ERK occurred at lower doses of LPS in monocytes from patients heterozygous and homozygous for CFTR mutations. These results indicate that a single allelic CFTR mutation is sufficient to augment IL-8 secretion in response to LPS. This is not a result of increased LPS receptor expression but, rather, is associated with alterations in MAPK signaling. PMID- 15358639 TI - Serological response to Pasteurella multocida NanH sialidase in persistently colonized rabbits. AB - Pasteurella multocida is a mucosal pathogen that colonizes the upper respiratory system of rabbits. Respiratory infections can result, but the bacteria can also invade the circulatory system, producing abscesses or septicemia. P. multocida produces extracellular sialidase activity, which is believed to augment colonization of the respiratory tract and the production of lesions in an active infection. Previously, it was demonstrated that some isolates of P. multocida contain two unique sialidase genes, nanH and nanB, that encode enzymes with different substrate specificities (S. Mizan, A. D. Henk, A. Stallings, M. Meier, J. J. Maurer, and M. D. Lee, J. Bacteriol. 182:6874-6883, 2000). We developed a recombinant antigen enzyme-linked immunosorbent assay (ELISA) based on the NanH sialidase of P. multocida and demonstrated that rabbits that were experimentally colonized with P. multocida produce detectable anti-NanH immunoglobulin M (IgM) and IgG in serum, although they demonstrated no clinical signs of pasteurellosis. In addition, clinically ill pet rabbits infected with P. multocida possessed IgM and/or IgG antibody against NanH. The NanH ELISA may be useful for the diagnosis of P. multocida infections in sick rabbits as well as for screening for carriers in research rabbit colonies. PMID- 15358640 TI - Regeneration and tolerance factor prevents bystander T-cell death associated with human immunodeficiency virus infection. AB - Human immunodeficiency virus (HIV) infection is characterized by a depletion of T cells. This depletion is caused both by the virus-induced death of infected T cells and by the death of uninfected cells (bystander depletion) by a mechanism which is largely uncharacterized. Regeneration and tolerance factor (RTF) is a subunit of the vacuolar ATPase and a protein that is involved with activation and apoptosis. Anti-RTF antibodies mediate apoptosis in T lymphocytes. When anti-RTF was added to lymphocytes from an HIV-positive individual, they underwent larger amounts of apoptosis than cells taken from healthy controls. When lymphocytes were examined by Western blotting, those from HIV-positive individuals exhibited increased levels of expression of the 50-kDa protein (P < 0.001). A 70-kDa protein was the predominant form of RTF in uninfected control lymphocytes, being expressed in 100% of individuals studied. The expression of the 50-kDa protein in HIV-positive individuals correlated with decreased absolute CD4 counts with a sensitivity of 92% and a positive predictive value of 86%. When uninfected lymphocytes were stimulated with anti-CD3 and anti-CD28, no RTF was detected during early stimulation but a 50-kDa protein was expressed during late stimulation. When the susceptibilities of the lymphocytes to anti-RTF-induced apoptosis were measured, they correlated with the size of the RTF protein expressed. The cells were not susceptible to apoptosis when the 70-kDa RTF was present but were susceptible when the 50-kDa RTF was present. We propose that the increase in the levels of the 50-kDa RTF on cells from HIV-positive individuals is important in preventing the cell from undergoing apoptosis. PMID- 15358641 TI - Measurement of immunoglobulin concentrations in the feces of healthy dogs. AB - Selective immunoglobulin A (IgA) deficiency is the most common primary immunodeficiency in humans and may be associated with chronic gastrointestinal disease. This observation has led to the suggestion that the high susceptibility of German shepherd dogs (GSD) to chronic enteropathies is related to a deficiency in mucosal IgA production. Relative deficiencies of IgA has been reported in the serum, saliva, tears, and feces of GSD both with and without alimentary disease; however, the findings of different studies are not consistent. The aim of this study was to confirm whether a relative deficiency of IgA exists in the feces of GSD. Feces were collected from healthy GSD (n = 209), Labrador retrievers (n = 96), beagles (n = 19), and miniature schnauzers (n = 32). Fecal IgA, IgM, and IgG were measured by capture enzyme-linked immunosorbent assays. Fecal IgG concentrations in the four breed groups were not significantly different. IgA concentrations were significantly greater in miniature schnauzers than in GSD (P = 0.0003) and Labradors (P = 0.0004) but not significantly different from those in beagles. IgM concentrations were significantly greater in miniature schnauzers than in GSD (P < 0.0001), Labradors (P < 0.0001), and beagles (P = 0.0098). These findings do not support the hypothesis that GSD have a relative deficiency in fecal IgA. The differences in immunoglobulin concentrations measured from a single defecation, between individuals of the same breed and between breeds, as well as the lack of an internal control molecule, make the determination of a normal reference range for all dogs impossible. Therefore, the usefulness of fecal immunoglobulin quantification for the assessment of intestinal immunoglobulin secretion in dogs is limited. PMID- 15358642 TI - Antigen recognition by serum antibodies in white-tailed deer (Odocoileus virginianus) experimentally infected with Mycobacterium bovis. AB - White-tailed deer (Odocoileus virginianus) have emerged as reservoirs of bovine tuberculosis in northern America. For tuberculosis surveillance of deer, antibody based assays are particularly attractive because deer are handled only once and immediate processing of the sample is not required. Sera collected sequentially from 25 Mycobacterium bovis-infected and 7 noninfected deer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for immunoglobulin specific to M. bovis antigens. Various routes of experimental M. bovis infection, such as intratonsillar inoculation (n = 11), aerosol (n = 6), and exposure to infected deer (in contact, n = 8), were studied. Upon infection, specific bands of reactivity at approximately 24 to 26 kDa, approximately 33 kDa, approximately 42 kDa, and approximately 75 kDa to M. bovis whole-cell sonicate were detected by immunoblot. Lipoarabinomannan-specific immunoglobulin was detected as early as 36 days postchallenge, and responses were detected for 94% of intratonsillarly and "in-contact"-infected deer. In MAPIA, sera were tested with 12 native and recombinant antigens coated on nitrocellulose. All in-contact-infected (8 of 8) and 10 of 11 intratonsillarly infected deer produced antibody reactive with one or more of the recombinant/native antigens. Responses were boosted by injection of tuberculin for intradermal tuberculin skin testing. Additionally, three of six deer receiving a very low dose of M. bovis via aerosol exposure produced antibody specific to one or more recombinant proteins. M. bovis was isolated from one of three nonresponding aerosol-challenged deer. Of the 12 antigens tested, the most immunodominant protein was MPB83; however, a highly sensitive serodiagnostic test will likely require use of multiple antigens. PMID- 15358643 TI - Effect of photopheresis on lymphocyte population in children with newly diagnosed type 1 diabetes. AB - In recent years photopheresis has been claimed to be an effective form of immunomodulation. It has also been shown to have an effect on the disease process at the onset of type 1 diabetes. In a double-blind, placebo-controlled randomized study, we analyzed if the effect of photopheresis in children with newly diagnosed diabetes is related to changes in the balance of lymhocyte populations. We also analyzed if lymphocyte subsets were related to recent infection, mild or aggressive disease manifestations, heredity, or gender. Nineteen children received active treatment with photopheresis, while 21 children received sham pheresis (placebo group). No influence of a history of previous infection, heredity, or certain clinical parameters on lymphocyte subsets was found. At the onset of type 1 diabetes, girls showed a higher proportion and a larger number of T cells (CD3+) and T-helper cells (CD4+) and a higher proportion of naive CD4+ CD45RA+ cells. In the placebo group, an increase in the number of subsets with the activated phenotype in both the CD4(CD29+) and the CD8 (CD11a+) compartments was noted during the course of the study. These changes did not occur in the photopheresis group. No relation between lymphocyte subsets and clinical outcome was found 1 year after the treatment with photopheresis. In conclusion, we found no major effect of photopheresis on lymphocyte populations in a group of children with newly diagnosed type 1 diabetes. However, in the placebo group the proportions of activated CD4 and CD8 cells increased over time. Since these changes did not occur in the actively treated group, our findings suggest that photopheresis may have some suppressive effects. PMID- 15358644 TI - Analysis of eight commercial enzyme immunoassay tests for detection of antibodies to Mycoplasma pneumoniae in human serum. AB - Mycoplasma pneumoniae is an important etiologic agent of primary atypical pneumonia in children and adults. The diagnosis of M. pneumoniae infection is commonly confirmed through serologic testing. In this study, we used paired sera from 51 patients (all with confirmed M. pneumoniae infection and positive complement fixation [CF] titers) to compare the results of eight enzyme immunoassays (EIAs) available commercially in the United States. We compared two single-use EIAs and six plate-type EIAs. Results from acute-phase sera ranged from only 7 (14%) positive by ImmunoWELL (GenBio) immunoglobulin M (IgM) EIA to 23 (45%) positive by Zeus IgG EIA. When both the acute-phase and convalescent phase serum samples were examined, positive results ranged from 20 (39%) by the ImmunoWELL (GenBio) IgM assay to 45 (88%) positive by the Remel IgG-IgM EIA. In this study, the single-use EIAs by Remel and Meridian were more reliable than were the plate-type EIAs. Among the plate-type EIAs, the Zeus and DiaSorin assays (which detect antibodies to protein antigens) were more sensitive than the ImmunoWELL assay (which detects antibodies to glycolipid antigens). In general, IgG EIAs on convalescent-phase sera were more concordant with one another than were IgM EIAs with one another. Scatter plot analysis of convalescent-phase sera showed that, as the CF titer dropped, the IgM assays identified fewer positive convalescent-phase sera. In contrast, the IgG assays provided fairly consistent positive results for convalescent-phase sera with CF titers of 64 and above. Results of individual tests and overall limitations of serodiagnostics for M. pneumoniae infections are discussed. PMID- 15358645 TI - Occurrence and potential diagnostic applications of serological cross reactivities between Brucella and other alpha-proteobacteria. AB - Agrobacterium, Sinorhizobium, and Ochrobactrum are genera closely related to Brucella but, in contrast to the latter, are not pathogenic for humans and animals. We studied by an indirect enzyme-linked immunosorbent assay (ELISA) the reactivities of brucellosis sera against cytosolic (CYT) and membrane (MA) antigens from these nonpathogenic bacteria, and we evaluated the potential usefulness of these cross-reactions for the diagnosis of brucellosis in humans, sheep, cows, and dogs. Canine infection by Brucella canis was detected with high specificity by CYT antigen-based ELISAs (96% for Agrobacterium, 96% for Sinorhizobium, and 91% for Ochrobactrum), while sensitivity was variable (58% for Agrobacterium, 88% for Sinorhizobium, and 84% for Ochrobactrum). In addition, it was possible to diagnose canine disease shortly after exposure to the pathogen (15 days). Similar results for canine brucellosis were obtained with MA antigens. In contrast, normal sera from humans, sheep, and cattle reacted strongly with all the antigens (CYT and MA antigens from the three bacteria), producing high cutoff values and, consequently, low sensitivities. While for some host species the reactivity patterns of normal sera by Western blotting were similar to those produced with sera from infected individuals, the reactivity pattern of bovine sera against Sinorhizobium meliloti antigens exhibited some differential bands for the two groups of sera. These results show that crude fractions from nonpathogenic alpha-proteobacteria can be used to diagnose canine brucellosis but may need to be further separated into simpler fractions to have diagnostic usefulness in ovine, bovine, or human infection. By reducing the biosafety requirements, the use of antigens derived from these nonpathogenic bacteria would simplify the production of diagnostic kits for brucellosis, especially in settings where biosafety level-3 facilities are scarce or absent. PMID- 15358646 TI - Analysis of amino acid sequence variations and immunoglobulin E-binding epitopes of German cockroach tropomyosin. AB - The allergenicities of tropomyosins from different organisms have been reported to vary. The cDNA encoding German cockroach tropomyosin (Bla g 7) was isolated, expressed, and characterized previously. In the present study, the amino acid sequence variations in German cockroach tropomyosin were analyzed in order to investigate its influence on allergenicity. We also undertook the identification of immunodominant peptides containing immunoglobulin E (IgE) epitopes which may facilitate the development of diagnostic and immunotherapeutic strategies based on the recombinant proteins. Two-dimensional gel electrophoresis and immunoblot analysis with mouse anti-recombinant German cockroach tropomyosin serum was performed to investigate the isoforms at the protein level. Reverse transcriptase PCR (RT-PCR) was applied to examine the sequence diversity. Eleven different variants of the deduced amino acid sequences were identified by RT-PCR. German cockroach tropomyosin has only minor sequence variations that did not seem to affect its allergenicity significantly. These results support the molecular basis underlying the cross-reactivities of arthropod tropomyosins. Recombinant fragments were also generated by PCR, and IgE-binding epitopes were assessed by enzyme-linked immunosorbent assay. Sera from seven patients revealed heterogeneous IgE-binding responses. This study demonstrates multiple IgE-binding epitope regions in a single molecule, suggesting that full-length tropomyosin should be used for the development of diagnostic and therapeutic reagents. PMID- 15358647 TI - Interleukin-15 enhances cytotoxicity, receptor expression, and expansion of neonatal natural killer cells in long-term culture. AB - Newborn infants have a higher susceptibility to various pathogens due to developmental defects in their host defense system, including deficient natural killer (NK) cell function. In this study, the effects of interleukin-15 (IL-15) on neonatal NK cells was examined for up to 12 weeks in culture. The cytotoxicity of fresh neonatal mononuclear cells (MNC) as assayed by K562 cell killing is initially much less than that of adult MNC but increases more than eightfold after 2 weeks of culture with IL-15 to a level equivalent to that of adult cells. This high level of cytotoxicity was maintained for up to 12 weeks. In antibody dependent cellular cytotoxicity (ADCC) assays using CEM cells coated with human immunodeficiency virus gp120 antigen, IL-15 greatly increased ADCC lysis by MNC from cord blood. IL-15 increased expression of the CD16+ CD56+ NK markers of cord MNC fivefold after 5 weeks of incubation. Cultures of neonatal MNC with IL-15 for up to 10 weeks resulted in a unique population of CD3- CD8+ CD56+ cells (more than 60%), which are not present in fresh cord MNC. These results show that IL-15 can stimulate neonatal NK cells and sustain their function for several weeks, which has implications for the clinical use of IL-15. PMID- 15358648 TI - Different methods of identifying new antigenic epitopes of human papillomavirus type 16 E6 and E7 proteins. AB - Human papillomavirus (HPV) infection is the most common cause of sexually transmitted viral infection and is the main cause of cervical cancer. Identification of HPV T-cell epitopes would be instrumental not only in our understanding of the protective immune response but also in the development of vaccines and immunotherapies. In contrast to viruses which cause systemic infection, identification of HPV epitopes is technically challenging because HPV causes a localized mucosal infection and the frequency of pathogen-specific T lymphocytes in peripheral blood is expected to be low. Here we describe three new antigenic epitopes (E7 7-15 [TLHEYMLDL], E6 52-61 [FAFRDLCIVY], and E7 79-87 [LEDLLMGTL]) of HPV 16 E6 and E7 proteins which have oncogenic activities. E7 7 15 was identified among peptides previously shown to bind to human leukocyte antigen (HLA)-A2.1 molecule, but it was found likely to be restricted by the HLA B48 molecule. E6 52-61 (likely to be restricted by HLA-B57) and E7 79-87 (likely to be restricted by HLA-B60) were detected, based on the magnitude of the T-cell immune responses, in another individual. In particular, T-cell clones specific for the E6 52-61 epitope were isolated effectively by magnetically selecting them based on gamma interferon secretion. This is an efficient method of identifying new epitopes of antigens for which the number of specific T lymphocytes in the circulation is expected to be small, and it should be widely applicable in identifying new T-cell epitopes. PMID- 15358649 TI - Prospective study of serological conversion as a risk factor for development of leprosy among household contacts. AB - Although the prevalence of leprosy has declined over the years, there is no evidence that incidence rates are falling. A method of early detection of those people prone to develop the most infectious form of leprosy would contribute to breaking the chain of transmission. Prophylactic treatment of serologically identified high-risk contacts of incident patients should be an operationally feasible approach for routine control programs. In addition, classification of high-risk household contacts will allow control program resources to be more focused. In this prospective study, we examined the ability of serology used for the detection of antibodies to phenolic glycolipid I of Mycobacterium leprae to identify those household contacts of multibacillary leprosy patients who had the highest risk of developing leprosy. After the start of multidrug therapy for the index case, a new case of leprosy developed in one in seven of the 178 households studied. In households where new cases appeared, the seropositivity rates were significantly higher (P < 0.001) than those in households without new cases. Seropositive household contacts had a significantly higher risk of developing leprosy (relative hazard adjusted for age and sex [aRH], 7.2), notably multibacillary leprosy (aRH = 24), than seronegative contacts. PMID- 15358650 TI - Increased production of proinflammatory cytokines following infection with porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae. AB - Induction of the proinflammatory cytokines interleukin-1 (IL-1) (alpha and beta), IL-6, IL-8, IL-10, IL-12, and tumor necrosis factor alpha (TNF-alpha) in pulmonary alveolar macrophages (PAMs) was assessed following experimental infection with porcine reproductive and respiratory syndrome virus (PRRSV) and/or Mycoplasma hyopneumoniae by using in vivo and in vitro models. The in vivo model consisted of pigs infected with PRRSV and/or M. hyopneumoniae and necropsied at 10, 28, or 42 days postinfection. Pigs infected with both pathogens had a greater percentage of macroscopic lung lesions, increased clinical disease, and slower viral clearance than pigs infected with either pathogen alone. The pigs infected with both PRRSV and M. hyopneumoniae had significantly increased levels of mRNA for many proinflammatory cytokines in PAMs collected by bronchoalveolar lavage (BAL) at all necropsy dates compared to those in uninfected control pigs. Increased levels of IL-1beta, IL-8, IL-10, and TNF-alpha proteins in BAL fluid, as measured by enzyme-linked immunosorbent assay, confirmed the increased cytokine induction induced by the pathogens. An in vitro model consisted of M. hyopneumoniae-inoculated tracheal ring explants cultured with PRRSV-infected PAMs. PAMs were harvested at 6 or 15 h postinfection with either or both pathogens. The in vitro study detected increased IL-10 and IL-12 mRNA levels in PAMs infected with PRRSV at all time periods. In addition, IL-10 protein levels were significantly elevated in the culture supernatants in the presence of M. hyopneumoniae-inoculated tracheal ring explants. The increased production of proinflammatory cytokines in vivo and in vitro associated with concurrent M. hyopneumoniae and PRRSV infection may play a role in the increased rates of pneumonia associated with PRRSV infection. The increased levels of IL-10 may be a possible mechanism that PRRSV and M. hyopneumoniae use to exacerbate the severity and duration of pneumonia induced by PRRSV and modulate the respiratory immune response. PMID- 15358651 TI - Comparison of oral fluid collectors for use in a rapid point-of-care diagnostic device. AB - Orally based diagnostic testing is emerging as an alternative, noninvasive method for analyzing a variety of analytes. These analytes include pathogens, antibodies, drugs, and nucleic acids. In the present study we developed a protocol for evaluation of collectors that could be used in orally based, point of-care diagnostics. A performance comparison was carried out with a number of commercially available collectors, and their ability to deliver fluid, proteins, bacteria, and nucleic acid from pathogens compatible with PCR was assessed. The collectors were all capable of picking up and delivering test materials, albeit at various levels. PMID- 15358652 TI - Differential immune responses to primary measles-mumps-rubella vaccination in Israeli children. AB - Measles remains an important cause of morbidity and mortality worldwide, primarily due to problems associated with delivery of the live attenuated vaccine to susceptible populations. In some developed countries, there is concern about the effects of immunization on the immune system. In this study, we analyzed the responses of 12-month-old Bedouin and Jewish children living in Israel to routine measles-mumps-rubella (MMR) vaccination. Seroconversion to measles was 99% in Bedouin and 79% in Jewish children (P < 0.01), and that to mumps and rubella was 92 to 100% in both groups. Measles neutralizing antibody titers were higher in Bedouin (333 +/- 39 mIU/ml) than Jewish (122 +/- 60 mIU/ml) children (P < 0.002). Immunoglobulin G levels were higher in Bedouin than Jewish children (P = 0.007) and increased after vaccination (P = 0.0009). Leukocyte (P < 0.02) and lymphocyte (P = 0.04) counts were higher and CD4 lymphocyte percentages were lower (P < 0.001) in Bedouin than Jewish children before and after vaccination. Leukocyte counts and natural killer cell numbers did not change after vaccination, but lytic activity increased in Bedouin children (P < 0.005). Spontaneous proliferation of cultured peripheral blood mononuclear cells increased with vaccination, but there were no changes in the proliferative responses to phytohemagglutinin or tetanus toxoid. In summary, no adverse effects of MMR vaccination on immune function were detected. However, there were differences in underlying immunologic parameters and in response to the measles component of the vaccine between Bedouin and Jewish children. It is not known whether genetic differences or environmental exposure accounts for these differences. PMID- 15358653 TI - Mass value assignment of total and subclass immunoglobulin G in a human standard anthrax reference serum. AB - An anti-Anthrax Vaccine Adsorbed (anti-AVA) standard human reference serum pool, AVR414, has been prepared, and the total and protective antigen (PA)-specific immunoglobulin G (IgG) were quantified. AVR414 was prepared by plasmapheresis of healthy adults who had received a minimum of four subcutaneous injections of AVA. Mass values (in milligrams per milliliter) for total IgG and IgG subclasses 1 to 4 were determined by radial immunodiffusion. Anti-PA-specific IgG assignment (in micrograms per milliliter) was done by consensus of two complementary approaches: homologous enzyme-linked immunosorbent assay (ELISA) with affinity-purified anti PA IgG as a calibrator and summation of mean PA-specific IgG subclass concentrations determined by IgG subclass-specific ELISA using the United States National Reference Preparation for Human Serum Proteins as a standard. The total IgG concentration assigned to AVR414 reference serum was 8.33 mg/ml. IgG subclass concentrations were the following: for IgG1, 4.48 mg/ml; for IgG2, 3.35 mg/ml; for IgG3, 0.37 mg/ml; and for IgG4, 0.30 mg/ml. The assigned mass value for total anti-PA-specific IgG was 141.2 microg/ml. Anti-PA-specific IgG subclass concentrations were the following: for IgG1, 79.6 microg/ml; for IgG2, 35.3 microg/ml; for IgG3, 3.2 microg/ml; and for IgG4, 25.3 microg/ml. Human reference serum pool AVR414 will have direct application in the standardization of anthrax serological assays, in reagent qualification, and as a standard for quantification of PA-specific IgG in humans who have been vaccinated with or otherwise exposed to Bacillus anthracis PA. PMID- 15358654 TI - Comparison of Western immunoblotting and the C6 Lyme antibody test for laboratory detection of Lyme disease. AB - Three commercial Lyme disease Western immunoblotting (WB) kits and the C6 Borrelia burgdorferi (Lyme) enzyme-linked immunosorbent assay (ELISA) kit were compared using two commercially available performance panels from the Centers for Disease Control and Prevention (CDC) and Boston Biomedica (BBI). Combined, the panels consisted of 52 characterized specimens. Immunoglobulin G (IgG) sensitivity was similar for the three WB products. The BBI and Marblot WBs were more specific for IgG antibodies, while the Virablot was the most sensitive for IgM antibody. The BBI WB was 100% specific for IgM, while Marblot was 97% and Virablot was 77% specific for IgM. The C6 ELISA was found to be 100% sensitive. Four false-positive C6 results were identified in patients that had clinically and microbiologically confirmed Lyme disease but were not detected by the CDC reference methods. No one WB product showed overall superiority. The C6 ELISA shows promise as the first ELISA for Lyme disease that would not require a supplemental test such as a WB. PMID- 15358655 TI - Low CD4+ T-lymphocyte values in human immunodeficiency virus-negative adults in Botswana. AB - CD4+-lymphocyte counts (LCs) play a crucial role in the management and monitoring of HIV infection. Variability in CD4+ LCs has been reported to occur as a result of measurement techniques and/or biological variations. We report on the CD4+ LCs of healthy human immunodeficiency virus (HIV)-seronegative adults in Botswana. Samples were obtained from HIV-seronegative blood donors. The median CD4+ LC was 726 cells/mm3 (for females, 782 cells/mm3; for males, 698 cells/mm3). The median CD8+ LC was 488 cells/mm3 (for females, 494 cells/mm3; for males, 485 cells/mm3). The median CD4+-to-CD8+ ratio was 1.57 (for females, 1.66; for males, 1.51). Our findings of low CD4+ LCs among HIV-negative adults in Botswana are significant and have important implications for the management of HIV disease in the population of this sub-Saharan African country. PMID- 15358656 TI - Leukotriene B4 receptor (BLT-1) modulates neutrophil influx into the peritoneum but not the lung and liver during surgically induced bacterial peritonitis in mice. AB - Leukotriene B4 (LTB4) is a rapidly synthesized, early neutrophil chemoattractant that signals via its cell surface receptor, BLT-1, to attract and activate neutrophils during peritonitis. BLT-1-deficient (BLT-1(-/-)) mice were used to determine the effects of LTB4 on neutrophil migration and activation, bacterial levels, and survival after cecal ligation and puncture (CLP). Male BLT-1(-/-) or wild-type (WT) BALB/c mice underwent CLP. Tissues were harvested for determination of levels of bacteria, myeloperoxidase (MPO), LTB4, macrophage inflammatory protein 2 (MIP-2), and neutrophil (polymorphonuclear leukocyte [PMN]) numbers at 4 and 18 h after CLP. PMN activation was determined by an assessment of phagocytosis ability and CD11b expression. Survival was also determined. BLT-1(-/-) mice had decreased numbers of PMNs in the peritoneum at both 4 and 18 h after CLP but increased numbers of PMNs in the blood at 18 h compared with WT mice. Liver and lung MPO levels were significantly higher in BLT 1(-/-) mice at both 4 and 18 h after CLP, with increased bacterial levels in the blood, the liver, and peritoneal fluid at 4 h. Bacterial levels remained higher in peritoneal fluid at 18 h, but blood and liver bacterial levels at 18 h were not different from levels at 4 h. PMN phagocytosis and CD11b levels were decreased in BLT-1(-/-) mice. LTB4 levels were similar between the groups before and after CLP, but MIP-2 levels were decreased both locally and systemically in BLT-1(-/-) mice. Survival was significantly improved in BLT-1(-/-) mice (71%) compared with WT mice (14%) at 48 h post-CLP. Thus, LTB4 modulates neutrophil migration into the mouse peritoneum, but not the lung or liver, after CLP. Despite higher bacterial and PMN levels at remote sites, there was increased survival in BLT-1(-/-) mice compared to WT mice. Decreased PMN activation may result in less remote organ dysfunction and improved survival. PMID- 15358657 TI - Epitope-specific antibody levels demonstrate recognition of new epitopes and changes in titer but not affinity during treatment of tuberculosis. AB - Antibody levels rise during treatment of tuberculosis. This study examined when this rise occurred, whether there was recognition of new antigen binding sites (epitopes) on the same or different antigens, and how long specific antibody persisted. Forty patients with smear-positive pulmonary tuberculosis provided serum before and during treatment. Antibody levels were measured using a monoclonal antibody competition assay to epitopes restricted to the Mycobacterium tuberculosis complex and an enzyme-linked immunosorbent assay for lipoarabinomannan. Significant increases in antibody levels were apparent after 7 days of treatment. Five samples (12.5%) had positive titers to all epitopes at the start of treatment, and this increased to 23 (58%) during treatment. Antibody to epitopes with the poorest sensitivity (the TB23 epitope of the 19-kDa antigen and the TB78 epitope of hsp65) showed the greatest increases after treatment. Antibody to these two epitopes was also absent in some patients with relapsed tuberculosis until after treatment. Antibody titers showed a biphasic response, with a fall at 2 to 3 months of treatment. Sera from two patients showed changes in the affinity of epitope-specific antibody during treatment, whereas the majority did not. Those infected with isoniazid-resistant strains of M. tuberculosis showed a late rise in antibody. Antibody to the TB68 epitope of the 16-kDa alpha-crystallin homolog was short-lived, but it recurred with bacteriological relapse during treatment. Positive antibody titers persisted for at least 3 to 18 months after treatment. Diagnostic tests for tuberculosis should be evaluated using only pretreatment sera. Delayed antigenic recognition could be due to active suppression and/or failure to engage internal antigens of M. tuberculosis. PMID- 15358658 TI - Analysis of possible factors affecting the specificity of the gamma interferon test in tuberculosis-free cattle herds. AB - Bovine tuberculosis (TB) is still a zoonotic problem in the world. Despite the fact that eradication programs for bovine TB are being implemented in many countries, it remains a public health problem. These programs are mainly based on a single intradermal tuberculin test using bovine tuberculin purified protein derivative (PPD), isolation, and slaughtering of infected animals. The aim of this study was to assess the specificity of the gamma interferon (IFN-gamma) test in TB-free cattle herds, by using not only Australian tuberculins but also tuberculins produced at our institute, and to correlate the response with the type of production (beef cattle, dairy cattle, and a dual-purpose breed), the housing system, and the age of the animals. We studied 800 animals selected from 20 TB- and paratuberculosis-free herds. The animals were tested in parallel, after stimulation with Australian tuberculins and tuberculins produced at our institute, by using the skin test and two IFN-gamma assays. The results of this trial showed that the specificity of the IFN-gamma test is higher than that of the skin test (96.8%) and ranges from 97.3% (using only Australian tuberculins) to 98.6% (using tuberculins produced at our institute). We found that different categories of cattle could influence the specificity of the skin test but that these differences tended to be reduced in the IFN-gamma assay, especially when Italian PPDs were used. PMID- 15358659 TI - Response to superantigen stimulation in peripheral blood mononuclear cells from children perinatally infected with human immunodeficiency virus and receiving highly active antiretroviral therapy. AB - Our understanding of the pathogenesis of perinatal human immunodeficiency virus (HIV) infection is still evolving. We sought to characterize the response to the bacterial superantigen Staphylococcus enterotoxin B (SEB) of lymphocytes from HIV infected children receiving treatment with highly active antiretroviral therapy (HAART). Using the flow cytometric methodology, we quantified apoptosis, proliferation, cytokine production, and activation antigen upregulation in CD4 and CD8 T lymphocytes following in vitro stimulation of peripheral blood mononuclear cells (PBMCs) with SEB. The levels of proliferation, CD4 interleukin 2 (IL-2) production, CD8 gamma interferon (IFN-gamma) production, and upregulation of CD69 expression by cells from HIV-infected children were indistinguishable from those by cells from controls. However, stimulation with SEB dramatically decreased the ratio of resting apoptotic cells to cycling apoptotic cells in the controls but not in the patients. In addition, unstimulated spontaneous apoptosis of CD4 T cells remained greater in the patients than in the controls. The percentages of IL-2-positive CD8 T cells and IFN-gamma-positive CD4 T cells following SEB stimulation were significantly lower in the patients than in the controls. Our multiparameter approach was able to demonstrate differences in lymphocyte superantigen responsiveness in HIV-infected children receiving HAART in comparison to that in uninfected controls, notably, an apoptotic versus a proliferative response to stimulation. PMID- 15358660 TI - CXCR2 blockade influences Anaplasma phagocytophilum propagation but not histopathology in the mouse model of human granulocytic anaplasmosis. AB - Anaplasma phagocytophilum is an obligate intracellular bacterium that infects neutrophils and causes human granulocytic anaplasmosis. Infection induces neutrophil secretion of interleukin-8 or murine homologs and perpetuates infection by recruiting susceptible neutrophils. We hypothesized that antibody blockade of CXCR2 would decrease A. phagocytophilum tissue load by interrupting neutrophil recruitment but would not influence murine hepatic pathology. C3H-scid mice were treated with CXCR2 antiserum or control prior to or on day 14 after infection. Quantitative PCR and immunohistochemistry for A. phagocytophilum were performed and severity of liver histopathology was ranked. Control mice had more infected cells in tissues than the anti-CXCR2-treated group. The histopathological rank was not different between treated and control animals. Infected cells of control mice clustered in tissue more than in treated mice. The results support the hypothesis of bacterial propagation through chemokine induction and confirm that tissue injury is unrelated to A. phagocytophilum tissue load. PMID- 15358661 TI - Membrane-anchored CD14 is important for induction of interleukin-8 by lipopolysaccharide and peptidoglycan in uroepithelial cells. AB - We investigated the induction of interleukin-8 (IL-8) by bacterial lipopolysaccharide (LPS) and peptidoglycan (PGN) in the bladder cancer cell lines T24, 5637, UM-UC-3, and HT1197. T24 and 5637 cells strongly induced IL-8 after stimulation with LPS or PGN in a dose- and time-dependent manner, whereas UM-UC-3 and HT1197 cells did so very weakly. The expression of CD14 at the mRNA, total cellular protein, and cell surface protein levels differed among these cell lines, but the expression levels of Toll-like receptors 2 and 4 (TLR2 and TLR4) were not significantly different. The CD14 expression levels were found to correlate with the inducibility of IL-8 by LPS or PGN. Treatment of T24 and 5637 cells with phosphatidylinositol-specific phospholipase C to eliminate CD14 from the cell surface dramatically suppressed the induction of IL-8. On the other hand, UM-UC-3 cells transfected with CD14 cDNA expressed membrane-anchored CD14 and showed more efficient induction of IL-8 by LPS stimulation than untransfected controls. These results suggest that the presence of the membrane-anchored, but not the soluble, form of CD14 is a strong factor in IL-8 induction in bladder epithelial cells in response to bacterial components. The presence of the membrane-anchored form of CD14 may thus be a determinant for the inflammatory response of uroepithelial cells. PMID- 15358662 TI - Antigen-specific gene expression profiles of peripheral blood mononuclear cells do not reflect those of T-lymphocyte subsets. AB - Advances in microarray technology have allowed for the monitoring of thousands of genes simultaneously. This technology is of particular interest to immunologists studying infectious diseases, because it provides tremendous potential for investigating host-pathogen interactions at the level of immune gene expression. To date, many studies have focused either on cell lines, where the physiological relevance is questionable, or on mixed cell populations, where the contributions of individual subpopulations are unknown. In the present study, we perform an intrasubject comparison of antigen-stimulated immune gene expression profiles between a mixed population of peripheral blood mononuclear cells (PBMC) and the two predominant cell types found in PBMC, CD4+ and CD8+ T lymphocytes. We show that the microarray profiles of CD4+ and CD8+ T lymphocytes differ from each other as well as from that of the mixed cell population. The independence of the gene expression profiles of different cell types is demonstrated with a ubiquitous antigen (Candida albicans) as well as with a disease-specific antigen (human immunodeficiency virus p24). This study has important implications for microarray studies of host immunity and underscores the importance of profiling the expression of specific cell types. PMID- 15358663 TI - Age-specific immune response to HspA in Helicobacter pylori-positive persons in Mexico. AB - The immune response to heat shock protein A (HspA) in Helicobacter pylori positive adults increases with age in developed countries. This response has not been studied with children or in developing countries (G. I. Perez-Perez, J. M. Thiberge, A. Labigne, and M. J. Blaser, J. Infect. Dis. 174:1046-1050, 1996). As determined by using a specific enzyme-linked immunosorbent assay, HspA seropositivity among 592 individuals in Mexico was <10% in children and increased to >40% in adults. PMID- 15358664 TI - Immune cell Toll-like receptor 4 is required for cardiac myocyte impairment during endotoxemia. AB - The aim of this study was to investigate the importance of Toll-like receptor 4 (TLR4) signaling on cardiac myocytes versus immune cells in lipopolysaccharide (LPS)-induced cardiac dysfunction. Cardiac myocytes isolated from LPS-treated C57Bl/6 mice showed reduced shortening and calcium transients as compared with myocytes from untreated mice. In addition, LPS-treated C57Bl/6 mice showed impaired cardiac mitochondrial function, including reduced respiration and reduced time of induction of permeability transition. All of the aforementioned cardiac dysfunction was dependent on TLR4, because LPS-treated TLR4-deficient mice did not have reduced myocyte shortening or mitochondrial dysfunction. To evaluate the role of cardiac myocyte versus leukocyte TLR4, LPS was injected into chimeric mice with TLR4-positive leukocytes and TLR4-deficient myocytes. These mice showed reduced myocyte shortening in response to LPS. Myocytes from chimeric mice with TLR4-deficient leukocytes and TLR4-positive myocytes had no response to LPS. In addition, isolated myocytes from C57Bl/6 mice subsequently treated with LPS and serum for various times did not have reduced shortening, despite the presence of TLR4 mRNA and protein, as determined by reverse-transcription polymerase chain reaction and fluorescent-activated cell sorting. In fact, cardiac myocytes had equivalent amounts of TLR4 as endothelium; however, only the latter is responsive to LPS. Furthermore, signaling pathways downstream of TLR4 were not activated during direct LPS treatment of myocytes. In conclusion, TLR4 on leukocytes, and not on cardiac myocytes, is important for cardiac myocyte impairment during endotoxemia. PMID- 15358665 TI - Experimental and clinical regenerative capability of human bone marrow cells after myocardial infarction. AB - Bone marrow mononuclear cells (BMCs) from 20 patients with extensive reperfused myocardial infarction (MI) were used to assess their myocardial regenerative capability "in vitro" and their effect on postinfarction left ventricular (LV) remodeling. Human BMCs were labeled, seeded on top of cryoinjured mice heart slices, and cultured. BMCs showed tropism for and ability to graft into the damaged mouse cardiac tissue and, after 1 week, acquired a cardiomyocyte phenotype and expressed cardiac proteins, including connexin43. In the clinical trial, autologous BMCs (78+/-41x10(6) per patient) were intracoronarily transplanted 13.5+/-5.5 days after MI. There were no adverse effects on microvascular function or myocardial injury. No major cardiac events occurred up to 11+/-5 months. At 6 months, magnetic resonance showed a decrease in the end systolic volume, improvement of regional and global LV function, and increased thickness of the infarcted wall, whereas coronary restenosis was only 15%. No changes were found in a nonrandomized contemporary control group. Thus, BMCs are capable of nesting into the damaged myocardium and acquire a cardiac cell phenotype in vitro as well as safely benefiting ventricular remodeling in vivo. Large-scale randomized trials are needed now to assess the clinical efficacy of this treatment. PMID- 15358666 TI - Reversible connexin 43 dephosphorylation during hypoxia and reoxygenation is linked to cellular ATP levels. AB - Altered gap junction coupling of cardiac myocytes during ischemia may contribute to development of lethal arrhythmias. The phosphoprotein connexin 43 (Cx43) is the major constituent of gap junctions. Dephosphorylation of Cx43 and uncoupling of gap junctions occur during ischemia, but the significance of Cx43 phosphorylation in this setting is unknown. Here we show that Cx43 dephosphorylation in synchronously contracting myocytes during ischemia is reversible, independent of hypoxia, and closely associated with cellular ATP levels. Cx43 became profoundly dephosphorylated during hypoxia only when glucose supplies were limited and was completely rephosphorylated within 30 minutes of reoxygenation. Similarly, direct reduction of ATP by various combinations of metabolic inhibitors and by ouabain was closely paralleled by loss of phosphoCx43 and recovery of phosphoCx43 accompanied restoration of ATP. Dephosphorylation of Cx43 could not be attributed to hypoxia, acid pH or secreted metabolites, or to AMP-activated protein kinase; moreover, the process was selective for Cx43 because levels of phospho-extracellular signal regulated kinase (ERK)1/2 were increased throughout. Rephosphorylation of Cx43 was not dependent on new protein synthesis, or on activation of protein kinases A or G, ERK1/2, p38 mitogen activated protein kinase, or Jun kinase; however, broad-spectrum protein kinase C inhibitors prevented Cx43 rephosphorylation while also sensitizing myocytes to reoxygenation-mediated cell death. We conclude that Cx43 is reversibly dephosphorylated and rephosphorylated during hypoxia and reoxygenation by a novel mechanism that is sensitive to nonlethal fluctuations in cellular ATP. The role of this regulated phosphorylation in the adaptation to ischemia remains to be determined. PMID- 15358667 TI - Targeted disruption of cd73/ecto-5'-nucleotidase alters thromboregulation and augments vascular inflammatory response. AB - To investigate the role of adenosine formed extracellularly in vascular homeostasis, mice with a targeted deletion of the cd73/ecto-5'-nucleotidase were generated. Southern blot, RT-PCR, and Western blot analysis confirmed the constitutive knockout. In vivo analysis of hemodynamic parameters revealed no significant differences in systolic blood pressure, ejection fraction, or cardiac output between strains. However, basal coronary flow measured in the isolated perfused heart was significantly lower (-14%; P<0.05) in the mutant. Immunohistochemistry revealed strong CD73 expression on the endothelium of conduit vessels in wild-type (WT) mice. Time to carotid artery occlusion after ferric chloride (FeCl3) was significantly reduced by 20% in cd73-/- mice (P<0.05). Bleeding time after tail tip resection tended to be shorter in cd73-/- mice (-35%). In vivo platelet cAMP levels were 0.96+/-0.46 in WT versus 0.68+/ 0.27 pmol/106 cells in cd73-/- mice (P<0.05). Under in vitro conditions, platelet aggregation in response to ADP (0.05 to 10 micromol/L) was undistinguishable between the two strains. In the cremaster model of ischemia-reperfusion, the increase in leukocyte attachment to endothelium was significantly higher in cd73 /- compared with WT littermates (WT 98% versus cd73-/- 245%; P<0.005). The constitutive adhesion of monocytes in ex vivo-perfused carotid arteries of WT mice was negligible but significantly increased in arteries of cd73-/- mice (P<0.05). Thus, our data provide the first evidence that adenosine, extracellularly formed by CD73, can modulate coronary vascular tone, inhibit platelet activation, and play an important role in leukocyte adhesion to the vascular endothelium in vivo. PMID- 15358668 TI - Neural crest cell plasticity and its limits. AB - The neural crest (NC) yields pluripotent cells endowed with migratory properties. They give rise to neurons, glia, melanocytes and endocrine cells, and to diverse 'mesenchymal' derivatives. Experiments in avian embryos have revealed that the differentiation of the NC 'neural' precursors is strongly influenced by environmental cues. The reversibility of differentiated cells (such as melanocytes or glia) to a pluripotent precursor state can even be induced in vitro by a cytokine, endothelin 3. The fate of 'mesenchymal' NC precursors is strongly restricted by Hox gene expression. In this context, however, facial skeleton morphogenesis is under the control of a multistep crosstalk between the epithelia (endoderm and ectoderm) and NC cells. PMID- 15358669 TI - The role of SEUSS in auxin response and floral organ patterning. AB - Genetic and physiological analyses implicate auxin flux in patterning, initiation and growth of floral organs. Within the Arabidopsis flower, the ETTIN/ARF3 transcription factor responds to auxin to effect perianth organ number and reproductive organ differentiation. This work describes a modifier of ettin that causes filamentous, mispositioned outer whorl organs and reduced numbers of malformed stamens in the double mutant. The modifier was discovered to be a new allele of the seuss (seu) mutant. SEU encodes a novel protein that is predicted to transcriptionally co-repress the AGAMOUS floral organ identity gene. The effects of seu on ett are shown to be independent of the SEU-AG pathway. Furthermore, morphological, physiological and genetic evidence implicate SEU in auxin-regulated growth and development. seu has a pleiotropic phenotype that includes reductions in several classic auxin responses such as apical dominance, lateral root initiation, sensitivity to exogenous auxin and activation of the DR5 auxin response reporter. seu displays a synergistic interaction with the auxin response mutant pinoid, producing flowers with few outer whorl organs. Collectively, these data suggest that SEU is a novel factor affecting auxin response. A model is proposed in which SEU functions jointly with ETT in auxin response to promote floral organ patterning and growth. PMID- 15358670 TI - Sp8 and Sp9, two closely related buttonhead-like transcription factors, regulate Fgf8 expression and limb outgrowth in vertebrate embryos. AB - Initiation and maintenance of signaling centers is a key issue during embryonic development. The apical ectodermal ridge, a specialized epithelial structure and source of Fgf8, is a pivotal signaling center for limb outgrowth. We show that two closely related buttonhead-like zinc-finger transcription factors, Sp8 and Sp9, are expressed in the AER, and regulate Fgf8 expression and limb outgrowth. Embryological and genetic analyses have revealed that Sp8 and Sp9 are ectodermal targets of Fgf10 signaling from the mesenchyme. We also found that Wnt/beta catenin signaling positively regulates Sp8, but not Sp9. Overexpression functional analyses in chick unveiled their role as positive regulators of Fgf8 expression. Moreover, a dominant-negative approach in chick and knockdown analysis with morpholinos in zebrafish revealed their requirement for Fgf8 expression and limb outgrowth, and further indicate that they have a coordinated action on Fgf8 expression. Our study demonstrates that Sp8 and Sp9, via Fgf8, are involved in mediating the actions of Fgf10 and Wnt/beta-catenin signaling during vertebrate limb outgrowth. PMID- 15358671 TI - Dpp gradient formation by dynamin-dependent endocytosis: receptor trafficking and the diffusion model. AB - Developing cells acquire positional information by reading the graded distribution of morphogens. In Drosophila, the Dpp morphogen forms a long-range concentration gradient by spreading from a restricted source in the developing wing. It has been assumed that Dpp spreads by extracellular diffusion. Under this assumption, the main role of endocytosis in gradient formation is to downregulate receptors at the cell surface. These surface receptors bind to the ligand and thereby interfere with its long-range movement. Recent experiments indicate that Dpp spreading is mediated by Dynamin-dependent endocytosis in the target tissue, suggesting that extracellular diffusion alone cannot account for Dpp dispersal. Here, we perform a theoretical study of a model for morphogen spreading based on extracellular diffusion, which takes into account receptor binding and trafficking. We compare profiles of ligand and surface receptors obtained in this model with experimental data. To this end, we monitored directly the pool of surface receptors and extracellular Dpp with specific antibodies. We conclude that current models considering pure extracellular diffusion cannot explain the observed role of endocytosis during Dpp long-range movement. PMID- 15358672 TI - Coexpression of corticotropin-releasing hormone and urotensin i precursor genes in the caudal neurosecretory system of the euryhaline flounder (Platichthys flesus): a possible shared role in peripheral regulation. AB - CRH and urotensin I (UI) are neuroendocrine peptides that belong to the superfamily of corticotropin-releasing factors. In mammals, these peptides regulate the stress response and other central nervous system functions, whereas in fish an involvement for UI in osmoregulation has also been suggested. We have identified, characterized, and localized the genes encoding these peptides in a unique fish neuroendocrine organ, the caudal neurosecretory system (CNSS). The CRH and UI precursors, isolated from a European flounder CNSS library, consist of 168 and 147 amino acid residues, respectively, with an overall homology of approximately 50%. Both precursors contain a signal peptide, a divergent cryptic region and a 41-amino acid mature peptide with cleavage and amidation sites. Genomic organization showed that whole CRH and UI coding sequences are contained in a single exon. Northern blot analysis and quantitative PCR of a range of tissues confirmed the CNSS as a major site of expression of both CRH and UI and thus serves as a likely source of circulating peptides. In situ hybridization demonstrated that CRH and UI colocalize to the same cells of the CNSS. Our findings suggest that, in euryhaline fish, the CNSS is a major site of production of CRH and probably contributes to the high circulating levels observed in response to specific environmental challenges. Furthermore, the localization of CRH and UI within the same cell population suggests an early, possibly shared role for these peptides in controlling stress-mediated adaptive plasticity. PMID- 15358673 TI - Differential signal transduction of progesterone and medroxyprogesterone acetate in human endothelial cells. AB - The conjugated equine estrogens-only arm of the Women's Health Initiative trial, showing a trend toward protection from heart disease as opposed to women receiving also medroxyprogesterone acetate (MPA), strengthens the debate on the cardiovascular effects of progestins. We compared the effects of progesterone (P) or MPA on the synthesis of nitric oxide and on the expression of leukocyte adhesion molecules, characterizing the signaling events recruited by these compounds. Although P significantly increases nitric oxide synthesis via transcriptional and nontranscriptional mechanisms, MPA is devoid of such effects. Moreover, when used together with physiological estradiol (E2) concentrations, P potentiates E2 effects, whereas MPA impairs E2 signaling. These findings are observed both in isolated human endothelial cells as well as in vivo, in ovariectomized rat aortas. A marked difference in the recruitment of MAPK and phosphatidylinositol-3 kinase explains the divergent effects of the two gestagens. In addition, both P and MPA decrease the adhesiveness of endothelial cells for leukocytes when given alone or with estrogen. MPA is more potent than P in inhibiting the expression of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1. However, when administered together with physiological amounts of glucocorticoids, MPA (which also binds glucocorticoid receptor) markedly interferes with the hydrocortisone-dependent stabilization of the transcription factor nuclear factor kappaB and with the expression of adhesion molecules, acting as a partial glucocorticoid receptor antagonist. Our findings show significant differences in the signal transduction pathways recruited by P and MPA in endothelial cells, which may have relevant clinical implications. PMID- 15358674 TI - Activation of the glucocorticoid receptor or liver X receptors interferes with growth hormone-induced akr1b7 gene expression in rat hepatocytes. AB - The akr1b7 gene encodes an aldo-keto reductase involved in detoxification of isocaproaldehyde, the product from side chain cleavage of cholesterol, and of 4 hydroxynonenal (4-HNE) formed by lipid peroxidation and cleavage. Here we show that the expression of akr1b7 mRNA in rat liver is sexually differentiated, expressed in females but not in males, and regulated by the sexually dimorphic secretion pattern of GH. A GH dose-dependent induction of akr1b7 was demonstrated in cultured primary rat hepatocytes, which was sensitive to cycloheximide. Activation of the glucocorticoid receptor (GR) or liver X receptors (LXR) by dexamethasone (Dex) and T1317, respectively, attenuated the GH-induced expression of akr1b7 and CYP2C12, the prototypical rat hepatic gene dependent on the female characteristic secretion pattern of GH. In contrast, neither Dex nor T1317 had any repressive effect on the GH induction of IGF-I mRNA. A common mechanism for LXR- and GR-mediated repressive actions on gene transcription is inhibition of nuclear factor (NF)-kappaB; however, EMSAs and pharmacological interference with NF-kappaB signaling provided no evidence for the involvement of NF-kappaB in the repressive action of Dex and T1317 on GH-induced akr1b7 expression. PMID- 15358675 TI - 16K-prolactin inhibits activation of endothelial nitric oxide synthase, intracellular calcium mobilization, and endothelium-dependent vasorelaxation. AB - Activation of endothelial nitric oxide synthase (eNOS) and subsequent nitric oxide production (NO) are events that mediate the effect of important angiogenic, vasopermeability, and vasorelaxation factors, including vascular endothelial growth factor (VEGF), bradykinin (BK), and acetylcholine (ACh). The N-terminal 16 kDa fragment of prolactin (16K-PRL) acts on endothelial cells to inhibit angiogenesis both in vivo and in vitro. Here, we show that 16K-PRL inhibits VEGF induced eNOS activation in endothelial cells. Inhibition of eNOS activation may mediate the antiangiogenic properties of 16K-PRL, because the NO donor (Z)-1-[2 (2-aminoethyl)- N-(2-ammonio-ethyl)amino]diazen-1-ium-1,2-diolate (DETANONOate) prevented 16K-PRL from blocking the VEGF-induced proliferation of endothelial cells. In addition, 16K-PRL inhibited eNOS activation by BK and blocked the BK evoked transient increase in intracellular Ca(2+) in endothelial cells. This finding suggests that 16K-PRL interferes with the mobilization of intracellular Ca(2+), thereby inhibiting the Ca(2+)-dependent activation of eNOS. Blockage of eNOS activation can lead to inhibition of vasodilation. Consistently, 16K-PRL inhibited BK-induced relaxation of coronary vessels in isolated perfused guinea pig hearts. Moreover, 16K-PRL inhibited eNOS activation induced by ACh, and this action resulted in the inhibition of both ACh-evoked relaxation of coronary vessels in isolated perfused rat hearts and ACh-induced, endothelium-dependent relaxation of rat aortic segments. In conclusion, 16K-PRL can block the Ca(2+) mediated activation of eNOS by three different vasoactive substances, and this action results in the inhibition of both angiogenesis and vasorelaxation. PMID- 15358676 TI - Regulation of 5alpha-reductase isoforms by oxytocin in the rat ventral prostate. AB - Oxytocin (OT) is present in the male reproductive tract, where it is known to modulate contractility, cell growth, and steroidogenesis. Little is known about how OT regulates these processes. This study describes the localization of OT receptor in the rat ventral prostate and investigates if OT regulates gene expression and/or activity of 5alpha-reductase isoforms I and II. The ventral prostates of adult male Wistar rats were collected following daily sc administration of saline (control), OT, a specific OT antagonist or both OT plus antagonist for 3 d. Expression of the OT receptor was identified in the ventral prostate by RT-PCR and Western blot, and confirmed to be a single active binding site by radioreceptor assay. Immunohistochemistry localized the receptor to the epithelium of prostatic acini and to the stromal tissue. Real-time RT-PCR determined that OT treatment significantly reduced expression of 5alpha-reductase I but significantly increased 5alpha-reductase II expression in the ventral prostate. Activity of both isoforms of 5alpha-reductase was significantly increased by OT, resulting in increased concentration of prostatic dihydrotestosterone. In conclusion, OT is involved in regulating conversion of testosterone to the biologically active dihydrotestosterone in the rat ventral prostate. It does so by differential regulation of 5alpha-reductase isoforms I and II. PMID- 15358677 TI - Characterization of crustacean cardioactive peptide as a novel insect midgut factor: isolation, localization, and stimulation of alpha-amylase activity and gut contraction. AB - The midgut, which plays a major role in the feeding behavior of insects, is believed to be controlled by various factors including neuropeptides. In the present study, we identified a neuropeptide crustacean cardioactive peptide (CCAP) as a novel gut factor in insects. CCAP was isolated from the midgut of the cockroach, Periplaneta americana. RT-PCR analysis detected the expression of the cockroach CCAP mRNA in the ventral nerve cord and the midgut, revealing the production of CCAP in the midgut as well as the central nervous system. Moreover, we observed expression of the CCAP gene in the endocrine cells of the midgut by in situ hybridization, and immunohistochemical analysis showed that CCAP was distributed around the lateral surfaces of the endocrine cells. Elevation of alpha-amylase activity was observed upon addition of CCAP to the midgut. These results suggest that CCAP is involved in digestion of carbohydrate in a paracrine manner. In situ hybridization and immunohistochemistry also revealed CCAP expression in the cell bodies of the ingluvial ganglion, which innervates the midgut muscle layer but not the epithelium, indicating that CCAP is produced in the ingluvial ganglion and then transported to the muscle layer through nerve fibers. Furthermore, CCAP was found to stimulate the contraction of the foregut, midgut, and hindgut in a dose-dependent manner. Taken together, our data indicate the multifunctionality of CCAP in the regulation of gut tissues as both a neuropeptide and a novel midgut factor. PMID- 15358678 TI - The carboxypeptidase E knockout mouse exhibits endocrinological and behavioral deficits. AB - A carboxypeptidase E (CPE) knockout (KO) mouse was generated by deletion of exons 4 and 5 from the CPE gene, and its phenotype was characterized. KO mice became obese by 10-12 wk of age and reached 60-80 g by 40 wk. At this age, body fat content was more than double that in the wild-type (WT) controls. The null animals consumed more food overall, were less physically active during the light phase of the light-dark cycle, and burned fewer calories as fat than WT littermates. Fasting levels of glucose and insulin-like immunoreactivity in plasma were elevated in both male and female KO mice at approximately 20 wk; males recovered fully and females partially from this state by 32 wk. At this time, insulin-like immunoreactivity in the plasma, identified as proinsulin, was 50-100 times higher than that of the WT animals. The KO mice showed impaired glucose clearance and were insulin resistant. High levels of leptin and no circulating fully processed cocaine- and amphetamine-related transcript, a peptide that is responsive to leptin-induced feedback inhibition of feeding, were found in serum. The KO mice were subfertile and showed deficits in GnRH processing in the hypothalamus. Behavioral analyses revealed that KO animals showed diminished reactivity to stimuli and had reduced muscle strength and coordination, as well as visual placing and toe-pinch reflexes. These data demonstrate that CPE KO mice display a wide range of neural and endocrine abnormalities and suggest that CPE may have additional physiological roles beyond those ascribed to peptide processing and sorting of prohormones in cells. PMID- 15358679 TI - Norepinephrine induction of mitogen-activated protein kinase phosphatase-1 expression in rat pinealocytes: distinct roles of alpha- and beta-adrenergic receptors. AB - In this study, we investigated the mechanisms through which norepinephrine (NE) regulates MAPK phosphatase-1 (MKP-1) expression in rat pinealocytes. Stimulation with NE (a mixed alpha- and beta-adrenergic agonist) caused a rapid increase in MKP-1 mRNA and protein that peaked around 1 h post stimulation, and the response was sustained for at least 4 h. Selective activation of beta-adrenergic receptors with isoproterenol for 1 h caused a similar increase in MKP-1 mRNA and protein as observed with NE, but at 3 h, the isoproterenol response was much lower relative to NE. In contrast, selective activation of alpha-adrenergic receptors caused only small increases in MKP-1 mRNA and protein and appeared to function primarily in prolonging the beta-adrenergic-stimulated responses. In NE-stimulated pinealocytes, blockade of beta-adrenergic receptors caused a rapid reduction in MKP-1 mRNA, but it had a minimal effect on MKP-1 protein. In contrast, blockade of alpha-adrenergic receptors specifically reduced NE-induced MKP-1 protein but not mRNA. At the postreceptor level, treatment with dibutyryl cAMP caused parallel increases in MKP-1 mRNA and protein. However, treatment with a protein kinase C activator caused a significant increase in MKP-1 protein but had little effect on MKP-1 mRNA. Together, these results suggest that, in rat pinealocytes, NE activates the beta-adrenergic receptor --> protein kinase A pathway to induce transcription and translation of MKP-1 expression and the alpha-adrenergic receptor --> protein kinase C pathway to prolong the stimulated responses through increased stability of the MKP-1 protein. PMID- 15358680 TI - Regulation of steroidogenesis and steroidogenic acute regulatory protein in R2C cells by DAX-1 (dosage-sensitive sex reversal, adrenal hypoplasia congenita, critical region on the X chromosome, gene-1). AB - In the present study, steroidogenesis in two different Leydig tumor cell lines was compared. One, the MA-10 mouse tumor cell line, produces steroids and the steroidogenic acute regulatory (StAR) protein only when stimulated by trophic hormones and cAMP analogs. The other, the R2C rat tumor cell line, produces steroids and the StAR protein constitutively without stimulation. We observed that high levels of DAX-1 (dosage-sensitive sex reversal, adrenal hypoplasia congenita, critical region on the X chromosome, gene-1), a repressor of steroidogenesis and StAR gene expression, were present in MA-10 cells but not in R2C cells. Based upon this observation, we hypothesized that the absence of DAX-1 might result in constitutive steroidogenesis in R2C cells. To test this hypothesis, DAX-1 was overexpressed in the R2C cells using the Tet-on inducible gene expression system and resulted in a 45% decrease in steroid production, a 35% decrease in StAR protein, and a 39% decrease in cytochrome P450 side chain cleavage expression. Further, using retroviral infection with DAX-1, StAR expression and steroidogenesis were decreased 50-60% and 60% in R2C cells, respectively. These results corroborate previous findings that DAX-1 negatively regulates steroid synthesis through the inhibition of StAR expression and indicate that the absence of DAX-1 in R2C cells is, at least in part, responsible for the constitutive steroidogenesis and StAR expression observed. PMID- 15358681 TI - Migration of antigen presenting cells from periphery to the peritoneum during an inflammatory response: role of chemokines and cytokines. AB - We demonstrate the migration of antigen presenting cells (APCs), macrophages, and dendritic cells from the subcutaneous site to the peritoneum after they have picked up the antigen, using cell tracking dye. The migration of the APCs is more universal as it was also observed after injection of MethA tumor, DH-5alpha cells, and leishmania parasites, in addition to AK-5 tumor cells. Cellular migration is mediated by several chemokines and cytokines that also induce heavy influx of immune cells into the peritoneum. MIP-3beta secreted by the mesothelial cells is involved in the cellular influx into the peritoneum, whereas IL-12 and IFN-gamma produced by the APCs induced activation of immune cells in the peritoneum. Our results suggest an antigen presentation function for the APCs in the peritoneum as studied by lymphoproliferation assays. These studies indicate antigen presentation function of the activated migratory APCs from the distant subcutaneous site to the peritoneum, suggesting it acts as an important lymphoid organ involved in the enhancement of effector cell function. PMID- 15358682 TI - The human DNA polymerase lambda interacts with PCNA through a domain important for DNA primer binding and the interaction is inhibited by p21/WAF1/CIP1. AB - In this paper we show that DNA polymerase lambda (pol lambda) interacts with proliferating cell nuclear antigen (PCNA) in vivo in human cells. Moreover, by using recombinant mutated PCNA, we could demonstrate that pol lambda interacts with both the interdomain-connecting loop and the nearby hydrophobic pocket on the anterior of PCNA and that critical residues within a helix-hairpin-helix domain of pol lambda, important for proper DNA primer binding, are also involved in the enzyme's interaction with PCNA. Finally, we show that the tumor suppressor protein p21(WAF1/CIP1) can efficiently compete in vitro with pol lambda for binding to PCNA. Given the high rate of frameshift mutations induced by pol lambda and its ability to bypass abasic sites, accurate regulation of pol lambda activity by PCNA and p21 concerted action might be important for preventing genetic instability. PMID- 15358683 TI - Proteomic analysis of hyperdynamic mouse hearts with enhanced sarcoplasmic reticulum calcium cycling. AB - Depressed sarcoplasmic reticulum (SR) Ca-cycling is a hallmark of human and experimental heart failure. Strategies to improve this impairment by either increasing SERCA2a levels or decreasing phospholamban (PLN) activity have been suggested as promising therapeutic targets. Indeed, ablation of PLN gene in mice was associated with greatly enhanced cardiac Ca-cycling and performance. Intriguingly, this hyperdynamic cardiac function was maintained throughout the lifetime of the mouse without observable pathological consequences. To determine the cellular alterations in the expression or modification of myocardial proteins, which are associated with the enhanced cardiac contractility, we performed a proteomics-based analysis of PLN knockout (PLN-KO) hearts in comparison to isogenic wild-types. By use of 2-dimensional gel electrophoresis (2 DE), approximately 3300 distinct protein spots were detected in either wild-type or PLN-KO ventricles. Protein spots observed to be altered between PLN-KO and wild-type hearts were subjected to tryptic peptide mass fingerprinting for identification by MALDI-TOF mass spectrometry in combination with LC/MS/MS analysis. In addition, two-dimensional 32P-autoradiography was performed to analyze the phosphorylation profiles of PLN-KO cardiomyocytes. We identified alterations in the expression level of more than 100 ventricular proteins, along with changes in phosphorylation status of important regulatory proteins in the PLN-KO. These protein changes were observed mainly in two subcellular compartments: the cardiac contractile apparatus, and metabolism/energetics. Our findings suggest that numerous alterations in protein expression and phosphorylation state occurred upon ablation of PLN and that a complex functional relationship among proteins involved in calcium handling, myofibrils, and energy production may exist to coordinately maintain the hyperdynamic cardiac contractile performance of the PLN-KO mouse in the long term. PMID- 15358684 TI - Controlling passive smoking through legislation in Ireland: an attack on civil liberty or good public health policy? PMID- 15358685 TI - Combination therapy for pulmonary arterial hypertension: still more questions than answers. PMID- 15358686 TI - The role of reference values in interpreting lung function tests. PMID- 15358687 TI - A new tool for an old disease. PMID- 15358688 TI - Interventional bronchoscopy for tuberculous tracheobronchial stenosis. AB - This study investigated the use of interventional bronchoscopic techniques in the management of patients with symptomatic tracheobronchial stenosis from tuberculosis. The current authors evaluated their experience with interventional bronchoscopic techniques in 21 consecutive patients at the Singapore General Hospital, Singapore, from November 1994 to March 2001. All patients underwent rigid bronchocopy using the Dumon rigid ventilating bronchosope under general anaesthesia. A combination of techniques was used (mechanical or balloon dilatation, Nd-YAG laser and stenting using the Dumon stent). The mean+/-SD increase in luminal diameter of the tracheal lesions was from 4.5+/-2.5 mm pre procedure to 11.9+/-1.7 mm post-procedure, whereas that for the mainstem bronchi stenosis was from 2.6+/-1.0 mm to 8.3+/-2.4 mm. All patients had immediate relief of symptoms post-intervention. Two patients who presented with acute respiratory failure could be weaned off mechanical ventilation immediately post-procedure. At the end of the study period, 52% (11 out of 21) remained asymptomatic. Bronchoscopic intervention provided immediate symptomatic relief in all of the studied patients. However, repeated sessions may be required to maintain this improvement. It is concluded that interventional bronchoscopic techniques are useful in the management of patients with tracheobronchial stenosis from tuberculosis. PMID- 15358689 TI - Endobronchial brachytherapy in the treatment of malignant lung tumours. AB - A prospective study was made to assess the short-term clinical and endoscopic response to high-dose-rate endobronchial brachytherapy (HDREB) in patients with malignant endobronchial tumours. From July 1995 to May 2000, 288 HDREB sessions were carried out on 81 patients. The mean patient age was 61.57 yrs (range 34 82); males were predominant (87.65%). Tumours were primary in 76 patients (93.82%) and metastatic in five patients (6.18%). The inclusion criteria were malignant endobronchial tumour and either palliative treatment for incurable disease or intent-to-cure treatment for residual malignancy on the bronchial resection surface after surgery or an inoperable tumour. The exclusion criteria were as follows: impediments to catheter placement, expected survival <2 months, Karnofsky index <60, or absence of informed consent. The clinical response of a symptom was categorised as complete (disappearance of the symptom), partial (less than complete) or absent. The endoscopic response was considered to be complete if lesions disappeared and biopsy findings remained negative 1 month after the last radiation session; partial if lesions improved to some extent, but the biopsy findings were positive; and absent if there was no change in relation to baseline. The technique consisted of delivering high-dose irradiation from an Ir192 source to a target volume using one or two endobronchial catheters inserted under optical or video bronchoscopic guidance. Four sessions were scheduled at weekly intervals and 500 cGy was applied per session over a length of 1-9 cm, measured 0.5-1 cm from the centre of the source. In total, 85% of the symptoms analysed (haemoptysis, cough, dyspnoea, expectoration, and stridor) disappeared with HDREB, which was categorised as a complete response. The endoscopic response was complete in 56.79% of patients, partial or less than complete in 40.74% and absent in 2.46%. One major complication occurred (bronchial fistula 1.2%), but no lethal haemoptysis. Minor complications (pneumonitis, bronchospasm and bronchial stenosis) each occurred in one patient (1.2%). High-dose-rate endobronchial brachytherapy is a good palliative treatment for endoluminal lung neoplasms, effectively alleviating symptoms and endoscopic evidence in many cases with an acceptable rate of complications. High-dose-rate endobronchial brachytherapy can be carried out as an intent-to-cure procedure in highly selected cases. PMID- 15358690 TI - Combination of bosentan with epoprostenol in pulmonary arterial hypertension: BREATHE-2. AB - The efficacy and safety of combining bosentan, an orally active dual endothelin receptor antagonist and epoprostenol, a continuously infused prostaglandin, in the treatment of pulmonary arterial hypertension (PAH) was investigated. In this double-blind, placebo-controlled prospective study, 33 patients with PAH started epoprostenol treatment (2 ng.kg(-1)min(-1) starting dose, up to 14+/-2 ng.kg( 1)min(-1) at week 16) and were randomised for 16 weeks in a 2:1 ratio to bosentan (62.5 mg b.i.d for 4 weeks then 125 mg b.i.d) or placebo. Haemodynamics, exercise capacity and functional class improved in both groups at week 16. In the combination treatment group, there was a trend for a greater (although nonsignificant) improvement in all measured haemodynamic parameters. There were four withdrawals in the bosentan/epoprostenol group (two deaths due to cardiopulmonary failure, one clinical worsening, and one adverse event) and one withdrawal in the placebo/epoprostenol group (adverse event). This study showed a trend but no statistical significance towards haemodynamics or clinical improvement due to the combination of bosentan and epoprostenol therapy in patients with pulmonary arterial hypertension. Several cases of early and late major complications were reported. Additional information is needed to evaluate the risk/benefit ratio of combined bosentan-epoprostenol therapy in pulmonary arterial hypertension. PMID- 15358691 TI - Regulation of interleukin-1beta and interleukin-1beta inhibitor release by human airway epithelial cells. AB - In asthma, human airway epithelial cells (HAECs) regulate the intensity of mucosal inflammation, in part, by releasing the pro-inflammatory cytokine interleukin (IL)-1beta. However, the IL-1beta inhibitors, IL-1 receptor antagonist (IL-1RA) and soluble IL-1 receptor type II (sIL-1RII), regulate IL 1beta bioactivity. In order to better understand the control of IL-1beta activity in the airway mucosa, the role(s) of tumour necrosis factor (TNF)-alpha, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in the release of IL-1beta and its inhibitors by cultured HAECs were examined. HAECs were treated with TNF-alpha (2-200 ng.mL(-1)), dibutyryl cAMP (0.01-1 mM), 8 bromo-cGMP (0.01-1 mM) or vehicle for 24 h, and cytokine levels in the HAEC conditioned medium were measured by enzyme-linked immunosorbent assay. HAECs produced IL-1beta, IL-1RA and sIL-1RII constitutively, but the inhibitor concentrations greatly exceeded that of IL-1beta (by approximately 100- and approximately 550-fold, respectively). TNF-alpha dose-dependently increased the levels of all IL-1beta cytokine family members. However, over the range of TNF alpha concentrations studied, IL-1beta concentration increased more than those of its inhibitors. cAMP increased constitutive and TNF-alpha-stimulated IL-1beta release but reduced that of sIL-1RII. In contrast, cGMP had no effect on IL-1beta but reduced IL-1RA and sIL-1RII release. Under basal conditions, the disproportionate release of inhibitors relative to interleukin-1beta by human airway epithelial cells probably prevents interleukin-1beta-mediated biological effects. Tumour necrosis factor-alpha, cyclic adenosine monophosphate and cyclic guanosine monophosphate may potentiate mucosal inflammation by increasing interleukin-1beta levels relative to those of its inhibitors in the airway mucosa. PMID- 15358692 TI - Inhaled IFN-gamma for persistent nontuberculous mycobacterial pulmonary disease due to functional IFN-gamma deficiency. AB - Pulmonary infection with nontuberculous mycobacteria (NTM) in previously healthy human immunodeficiency virus-seronegative individuals is difficult to treat. Recently, functional interferon (IFN)-gamma deficiency has been identified in individuals susceptible to this disease. Treatment with inhaled IFN-gamma for NTM pulmonary disease associated with functional IFN-gamma deficiency has not been previously described. In this study, the IFN-gamma pathway was characterised in an individual who had progressive NTM pulmonary infection, despite appropriate multidrug antibiotic therapy, and 10 healthy controls. Levels of IFN-gamma and tumour necrosis factor-alpha in whole blood were assessed before and after incubation with lipopolysaccharide, heat-killed Escherichia coli, heat-killed Staphylococcus aureus and phorbol myristate acetate/ionomycin. The coding regions of interleukin (IL)-12, IL-18 and the IL-12 receptor were sequenced using nested primers. IFN-gamma1b (100 microg.dose(-1)) was administered to the affected individual by ultrasonic nebuliser 3 days.week(-1) for 3 months. In vitro whole blood production of IFN-gamma with and without physiological stimuli was consistent with functional IFN-gamma deficiency in the affected individual. There was no evidence of mutation in the coding regions of IL-12p35, IL-12p40, IL 12Rbeta1 and IL-18 in the affected individual. Treatment with inhaled IFN-gamma resulted in rapid and sustained clearance of the organism from the airways and stabilisation of lung function. In conclusion, inhaled interferon-gamma can be effective for the treatment of nontuberculous mycobacteria pulmonary disease associated with functional interferon-gamma deficiency. PMID- 15358693 TI - BMPR2 mutations in pulmonary arterial hypertension with congenital heart disease. AB - The aim of the present study was to determine if patients with both pulmonary arterial hypertension (PAH), due to pulmonary vascular obstructive disease, and congenital heart defects (CHD), have mutations in the gene encoding bone morphogenetic protein receptor (BMPR)-2. The BMPR2 gene was screened in two cohorts: 40 adults and 66 children with PAH/CHD. CHDs were patent ductus arteriosus, atrial and ventricular septal defects, partial anomalous pulmonary venous return, transposition of the great arteries, atrioventicular canal, and rare lesions with systemic-to-pulmonary shunts. Six novel missense BMPR2 mutations were found in three out of four adults with complete type C atrioventricular canals and in three children. One child had an atrial septal defect and patent ductus arteriosus; one had an atrial septal defect, patent ductus arteriosus and partial anomalous pulmonary venous return; and one had an aortopulmonary window and a ventricular septal defect. Bone morphogenetic protein receptor 2 mutations were found in 6% of a mixed cohort of adults and children with pulmonary arterial hypertension/congenital heart defects. The current findings compliment recent reports in mouse models implicating members of the bone morphogenetic protein/transforming growth factor-beta pathway inducing cardiac anomalies analogous to human atrioventricular canals, septal defects and conotruncal congenital heart defects. The small number of patients studied and the ascertainment bias inherent in selecting for pulmonary arterial hypertension require further investigation. PMID- 15358694 TI - Safety of research bronchoscopy, biopsy and bronchoalveolar lavage in asthma. AB - Bronchoscopy with endobronchial biopsy (EBB) and/or bronchoalveolar lavage (BAL) has become an important research tool in asthma. A recent report has suggested audit and reporting of the safety of these procedures. A total of 159 asthmatic patients (84 males, 75 females), aged 18-52 (median 27) yrs, forced expiratory volume in one second 53-120 (median 88) % predicted, underwent 273 bronchoscopies in six clinical research studies. On 228 occasions, EBB and BAL were performed and, on 45 occasions, EBB was performed alone. On 48 occasions, bronchoscopy was performed 24 h post-allergen challenge. Adverse events occurred on 34 out of 273 occasions, none of which were following allergen challenge. Post-EBB and BAL, four patients developed pleuritic chest pain, shortness of breath and fever. A further two patients experienced pleuritic chest pain alone post-EBB/BAL. Bronchospasm or worsening of asthma symptoms occurred on 14 occasions, 13 post EBB/BAL and on one occasion post-EBB alone. Fever/flu-like symptoms were reported on nine occasions following EBB and BAL. One subject had haemoptysis post EBB/BAL, but required no intervention. In conclusion, bronchoscopy, endobronchial biopsy and bronchoalveolar lavage can be performed safely in asthmatic patients. Most of the complications were seen where bronchoalveolar lavage and endobronchial biopsy were both performed, suggesting that bronchoalveolar lavage accounts for most of the adverse events. PMID- 15358695 TI - Exercise-induced flow limitation, dynamic hyperinflation and exercise capacity in patients with bronchial asthma. AB - It is known that, in stable asthmatics at rest, tidal expiratory flow limitation (EFL) and dynamic hyperinflation (DH) are seldom present. This study investigated whether stable asthmatics develop tidal EFL and DH during exercise with concurrent limitation of maximal exercise work rate (WRmax). A total of 20 asthmatics in a stable condition and aged 32+/-13 yrs (mean+/-SD) with a forced expiratory volume in one second (FEV1) of 101+/-21% of the predicted value were studied. Only three patients exhibited an FEV1 below the normal limits. On a first visit, patients performed a symptom-limited incremental (20 W.min(-1)) bicycle exercise test. On the second visit, the occurrence of EFL (using the negative expiratory pressure technique) and DH (via reduction in inspiratory capacity) were assessed at rest and when cycling at 33, 66 and 90% of their predetermined WRmax. FEV1 was measured to detect exercise-induced asthma, 5 and 15 min after stopping exercise at 90% WRmax. Only one patient showed EFL at rest, whereas 13 showed EFL and DH during exercise. In these 13 asthmatics, exercise capacity was significantly reduced (WRmax 75+/-9% pred) compared to the seven non EFL patients (WRmax 95+/-13% pred). Moreover, a significant correlation of WRmax (% pred) to the change in inspiratory capacity (percentage of resting value) from rest to 90% WRmax was found. Tidal EFL during exercise was not associated with exercise-induced asthma, which was detected in only three patients. In conclusion, tidal expiratory flow limitation and dynamic hyperinflation during exercise are common in stable asthmatics with normal spirometric results and without exercise-induced asthma, and may contribute to reduction in exercise capacity. PMID- 15358696 TI - Dynamic hyperinflation and tolerance to interval exercise in patients with advanced COPD. AB - Dynamic hyperinflation (DH) contributes importantly to the limitation of constant load exercise (CLE) in patients with chronic obstructive pulmonary disease (COPD). However, its role in the limitation of interval exercise (IE) remains to be explored. The change (Delta) in inspiratory capacity (IC) was measured to reflect changes in DH in 27 COPD patients (forced expiratory volume in one second mean+/-SEM % predicted: 40+/-3) at the end of a symptom-limited CLE test at 80% of peak work capacity (WRmax) and an IE test at 100% WRmax (30 s of work, alternated with 30 s of unloaded pedalling). At the limit of tolerance in both IE and CLE, patients exhibited similar DH (DeltaIC: 0.39+/-0.05 L and 0.45+/-0.05 L, respectively). However, exercise endurance time (t end) for IE (32.7+/-3.0 min) was significantly greater than for CLE (10.3+/-1.6 min). The IE t end correlated with resting IC, expressed as % pred normal. At 30 and 90% of total IE t end, DeltaIC (0.43+/-0.06 and 0.39+/-0.05 L, respectively) and minute ventilation (31.1+/-1.6 and 32.7+/-2.2 L.min(-1), respectively) were not significantly different. Resting hyperinflation helps to explain the limitation of interval exercise. Implementation of interval exercise for rehabilitation should provide important clinical benefits because it prolongs exercise endurance time and allows sustaining higher stable ventilation. PMID- 15358697 TI - Smoking cessation improves both direct and indirect airway hyperresponsiveness in COPD. AB - Smoking induces chronic obstructive pulmonary disease (COPD) and is associated with airway inflammation and airway hyperresponsiveness (AHR). It has not been studied in COPD whether direct (methacholine) and indirect (adenosine-5' monophosphate (AMP)) stimuli are associated with airway inflammation and neither whether smoking cessation improves these features. The current authors cross sectionally investigated the relationship of AHR to methacholine and AMP with lung function and inflammatory cells in the sputum of 33 smokers with COPD. In addition, changes in these parameters were prospectively assessed in 14 smokers who successfully quit smoking for 1 yr. The presence of AHR to both methacholine and AMP was associated with lower lung function, but not with sputum inflammation. AHR to methacholine and AMP improved significantly after a 1-yr smoking cessation, yet this was unrelated to changes in sputum cell counts. The numbers of neutrophils and epithelial cells significantly increased with smoking cessation. Both direct and indirect airway hyperresponsiveness are associated with lower lung function, but not with sputum inflammation in chronic obstructive pulmonary disease. Interestingly, 1-yr smoking cessation improved both direct and indirect airway hyperresponsiveness, yet without a significant association with changes in lung function or sputum inflammation. Thus, other factors are likely to induce these improvements, e.g. a reduction in stimulation of irritant receptors, airway wall changes or mucus hypersecretion. PMID- 15358698 TI - Spirometric reference equations for European females and males aged 65-85 yrs. AB - The aim of this study was to describe spirometric reference equations for healthy never-smoking European adults aged 65-85 yrs and to compare the predicted values of this sample with those from other studies including middle-aged and/or older adults. Reference equations and normal ranges for forced expiratory volume in one second (FEV1), forced vital capacity (FVC), forced expiratory volume in six seconds (FEV6), FEV1/FVC ratio and FEV1/FEV6 ratio were derived from a healthy subgroup of 458 subjects aged 65-85 yrs. Spirometry examinations followed the 1994 American Thoracic Society recommendations and the quality of the data was continuously monitored and maintained. Reference values and lower limits of normal were derived using a piecewise polynomial model with age and height as predictors. The reference values of FEV1 and FVC from the present study were higher than those given by prediction equations from the European Community for Coal and Steel. By contrast, use of prediction equations from Caucasian-American elderly subjects (Cardiovascular Health Study) consistently overpredicted FVC and FEV1 in females by 8.5 and 2.1%, respectively. In males, equations from the Cardiovascular Health Study overpredicted FVC by 2.8%, whilst underpredicting FEV1 by 2.5%. In conclusion, these results underscore the importance of using prediction equations appropriate to the origin, age and height characteristics of the subjects being studied. PMID- 15358700 TI - Primary pulmonary lymphangiectasia in infancy and childhood. AB - Primary pulmonary lymphangiectasia (PPL) is a rare disorder of unknown aetiology characterised by dilatation of the pulmonary lymphatics. PPL is widely reported to have a poor prognosis in the neonatal period and little is known about the clinical features of patients who survive the newborn period. The current authors report the outcome in nine patients diagnosed in infancy with PPL over a 15-yr period at a single university-based hospital clinic and followed for a median of 6 yrs. Although all of the patients initially experienced respiratory distress, respiratory symptoms improved in most patients after infancy and were notably better by the age of 6 yrs. Many patients had poor weight gain in the first years of life, which eventually improved. Radiological scans showed progressive resolution of neonatal infiltrates, but were characterised by hyperinflation and increased interstitial markings in older children. Most patients had evidence of bronchitis and grew pathogenic organisms from quantitative bronchoalveolar lavage culture. Pulmonary function tests showed predominantly obstructive disease that did not deteriorate over time. In conclusion, these results suggest that primary pulmonary lymphangiectasia does not have as dismal a prognosis as previously described and symptoms and clinical findings improve after the first year of life. PMID- 15358701 TI - Side-effects of fluticasone in asthmatic children: no effects after dose reduction. AB - To assess long-term effects and side-effects of fluticasone propionate (FP), a 2 yr study was performed, comparing a step-down dose approach (1,000 microg.day( 1), with reductions every 2 months to 500, 200 and 100 microg.day(-1) for the remainder of the study) versus a constant dose (200 microg.day(-1)). In 55 children with chronic persistent asthma, aged 6-10 yrs, airways hyperresponsiveness (AHR) and systemic side-effects (height, bone parameters and adrenal cortical function) were assessed at predetermined intervals in a double blind prospective 2-yr study. AHR improved after 4 months treatment with 1,000 microg.day(-1) FP followed by 500 microg.day(-1), without significant differences during long-term treatment between the two approaches. Dose-dependent reduction of growth velocity, adrenal cortical function and biochemical bone turnover was found during therapy with 1,000 and 500 microg.day(-1) FP when compared with 200 microg.day(-1). In conclusion, doses of 1,000 and 500 microg.day(-1) fluticasone propionate are associated with marked reductions of growth velocity, bone turnover and adrenal cortical function. However, conventional doses (< or =200 microg.day(-1) fluticasone propionate) appear to be safe in the long-term management of childhood asthma. From a safety point of view, high doses of fluticasone propionate should only be prescribed in exceptions, e.g. in persistent severe asthma. PMID- 15358699 TI - Phase II of the International Study of Asthma and Allergies in Childhood (ISAAC II): rationale and methods. AB - International comparative studies, investigating whether disease incidence or prevalence rates differ between populations and, if so, which factors explain the observed differences, have made important contributions to the understanding of disease aetiology in many areas. In Phase I of the International Study of Asthma and Allergies in Childhood (ISAAC), the prevalence rates of symptoms of asthma, allergic rhinitis and atopic eczema in 13-14-yr-olds, assessed by standardised questionnaires, were found to differ >20-fold between the 155 study centres around the world. Phase II of ISAAC aims to identify determinants of these differences by studying informative populations. A detailed study protocol was developed for use in community-based random samples of children aged 9-11 yrs. The study modules include standardised questionnaires with detailed questions on the occurrence and severity of symptoms of asthma, allergic rhinitis and atopic eczema, their clinical management, and a broad range of previous and current exposure conditions. In addition, standardised protocols were applied for examination of flexural dermatitis, skin-prick testing, bronchial challenge with hypertonic saline, blood sampling for immunoglobulin E analyses and genotyping, and dust sampling for assessment of indoor exposures to allergens and endotoxin. To date, ISAAC II field work had been completed or started in 30 study centres in 22 countries. The majority of centres are in countries that participated in International Study of Asthma and Allergies in Childhood Phase I and reflect almost the full range of the observed variability in Phase I prevalence rates. PMID- 15358702 TI - Surfactant proteins in pulmonary alveolar proteinosis in adults. AB - Pulmonary alveolar proteinosis (PAP) is a rare disorder characterised histologically by an intra-alveolar accumulation of fine granular eosinophilic and periodic acid-Schiff positive material. In a retrospective study, the composition of the intra-alveolarly accumulated material of adult patients with PAP was analysed by means of immunohistochemistry and Western blotting. In patients with PAP, the current authors found an intra-alveolar accumulation of surfactant protein (SP)-A, precursors of SP-B, SP-B, variable amounts of mono-, di-, and oligomeric SP-C forms, as well as SP-D. Only in one patient was a precursor of SP-C detected. By means of immuno-electron microscopy, the current authors identified not only transport vesicles labelled for precursors of SP-B and SP-C, but also transport vesicles containing either precursors of SP-B or SP C in type-II pneumocytes in normal human lungs. It is concluded that pulmonary alveolar proteinosis in adults is characterised by an intra-alveolar accumulation of surfactant protein A, precursors of surfactant protein B, and surfactant proteins B, C and D. The current data provide evidence that not only an impairment of surfactant clearance by alveolar macrophages, but also an abnormal secretion of transport vesicles containing precursors of surfactant protein B (but not surfactant protein C) and an insufficient palmitoylation of surfactant protein C, which may lead to the formation of di- and oligomeric surfactant protein C forms, play a role in the pathogenesis of pulmonary alveolar proteinosis. PMID- 15358703 TI - Pulmonary function and exercise capacity in survivors of severe acute respiratory syndrome. AB - The aim of this study was to investigate pulmonary function and exercise capacity in a group of survivors of the severe acute respiratory syndrome (SARS). At 3 months after hospital discharge, 46 survivors of SARS underwent the following evaluation: spirometry, static lung volumes and carbon monoxide transfer factor (TL,CO). In total, 44 of these patients underwent cardiopulmonary exercise testing. No abnormalities were detected in the pulmonary function tests in 23 (50%) of the patients. Abnormalities of forced vital capacity (FVC), forced expiratory volume in one second (FEV1), FEV1/FVC and TL,CO were detected in seven (15%), 12 (26%), one (2%) and 18 (39%) patients, respectively. All of these abnormalities were mild except in one case. In 18 patients (41%), the maximum aerobic capacity was below the lower limit of the normal range. Breathing reserve was low in four patients and significant oxygen desaturation was detected in a further four patients. Comparison of the measured exercise capacity with resting pulmonary function tests showed many cases of discordance in impairment. In conclusion, pulmonary function defects were detected in half of the recovered severe acute respiratory syndrome patients 3 months after hospital discharge, but the impairment was mild in almost all cases. Many patients had reduced exercise capacity that cannot be accounted for by impairment of pulmonary function. PMID- 15358704 TI - Home respiratory polygraphy for diagnosis of sleep-disordered breathing in heart failure. AB - The present study assessed the usefulness of home cardiorespiratory polygraphy as an alternative to overnight polysomnography for the diagnosis of sleep-disordered breathing in heart failure. A total of 75 patients with chronic stable heart failure due to systolic dysfunction (left ventricular ejection fraction < or =45%) underwent polysomnography at the Virgen del Rocio Hospital and cardiorespiratory polygraphy at home. The two studies were performed in random order and within 30 days. The accuracy of polygraphical findings was determined by sensitivity and specificity obtained by area under the receiver operating characteristics (ROC) curves for apnoea-hypopnoea index (AHI) thresholds of > or =5, > or =10 and > or =15 in the polysomnography. The area under the ROC curve for AHI > or =5, > or =10 and > or =15 was 0.896, 0.907 and 0.862, respectively. The diagnostic accuracy of polygraphy compared with polysomnography ranged 78.6 84%, with sensitivities of 68.4-82.5%, and specificities of 88.6-97.8% for the different AHI thresholds. Altogether, 29 patients had an AHI > or =10 (central pattern 24, obstructive pattern 5). Sensitivity and specificity of home polygraphy for identifying the sleep-disordered breathing pattern was 100%. These data show that home cardiorespiratory polygraphy has a high sensitivity and specificity for the diagnosis of sleep-disordered breathing associated with heart failure. PMID- 15358705 TI - Cystic fibrosis patients and families support cross-infection measures. AB - A clonal strain of Pseudomonas aeruginosa (PA) was isolated in 1999 at the Royal Children's Hospital, Melbourne, Australia, after five unrelated children with cystic fibrosis (CF) died from severe lung disease aged <5 yrs. Subsequently, more than half of the patients in the clinic with PA were found to harbour this strain, and segregation measures were instituted at the hospital to prevent further spread. The aim of this study was to assess CF parent and patient responses to the segregation measures to determine overall support. A questionnaire was sent out to the families of 291 CF children treated at the centre. A 65% response rate was obtained. The majority of parents (85%) and patients > or=12 yrs old (63%) were positive about the segregation measures instituted. A total of 11% of parents and 25% of patients were unsure, and 4% of parents and 12% of children gave negative responses. Those who were not happy listed reasons such as concerns about the emotional impact of not socialising with other CF children, inconclusive evidence about person-person spread of infection and feelings of alienation created in the clinic by the separation. In conclusion, the majority of responding cystic fibrosis patients and their families understand and are supportive of infection control measures instituted at the Royal Children's Hospital, Melbourne, Australia. PMID- 15358706 TI - Chest wall kinematics and respiratory muscle action in ankylosing spondylitis patients. AB - No direct measurements of the pressures produced by the ribcage muscles, the diaphragm and the abdominal muscles during hyperventilation have been reported in patients with ankylosing spondylitis. Based on recent evidence indicating that abdominal muscles are important contributors to stimulation of ventilation, it was hypothesised that, in ankylosing spondylitis patients with limited ribcage expansion, a respiratory centre strategy to help the diaphragm function may involve coordinated action of this muscle with abdominal muscles. In order to validate this hypothesis, the chest wall response to a hypercapnic/hyperoxic rebreathing test was assessed in six ankylosing spondylitis patients and seven controls by combined analysis of: 1) chest wall kinematics, using optoelectronic plethysmography, this system is accurate in partitioning chest wall expansion into the contributions of the ribcage and the abdomen; and 2) respiratory muscle pressures, oesophageal, gastric and transdiaphragmatic (Pdi); the pressure/volume relaxation characteristics of both the ribcage and the abdomen allowed assessment of the peak pressure of both inspiratory and expiratory ribcage muscles, and of the abdominal muscles. During rebreathing, chest wall expansion increased to a similar extent in patients to that in controls; however, the abdominal component increased more and the ribcage component less in patients. Peak inspiratory ribcage, but not abdominal, muscle pressure was significantly lower in patients than in controls. End-inspiratory Pdi increased similarly in both groups, whereas inspiratory swings in Pdi increased significantly only in patients. No pressure or volume signals correlated with disease severity. The diaphragm and abdominal muscles help to expand the chest wall in ankylosing spondylitis patients, regardless of the severity of their disease. This finding supports the starting hypothesis that a coordinated response of respiratory muscle activity optimises the efficiency of the thoracoabdominal compartment in conditions of limited ribcage expansion. PMID- 15358707 TI - Can psychological factors help us to determine adherence to CPAP? A prospective study. AB - The present study objective was to establish whether pretreatment social cognitive variables may contribute to the explanation of variance in adherence to continuous positive airway pressure (CPAP) treatment for patients with obstructive sleep apnoea/hypopnoea syndrome (OSAHS). A total of 119 of 180 consecutive OSAHS patients were recruited to the study prior to initial CPAP titration. Patients completed psychological measures of health value, health locus of control (incorporating internality, chance, powerful others) and self efficacy prior to CPAP titration. Objective adherence data were measured by CPAP unit time clocks and collected at 3-month follow-up. Average nightly use was calculated over this period. Logistic regression of prospective predictors of adherence produced a model comprising psychological (health value, internality, powerful others), as well as clinical variables (Epworth score, body mass index, apnoea/hypopnoea index, CPAP pressure). This model explained 24% of the variance in CPAP use, and correctly identified 75% of adherers and 53% of nonadherers. Although the psychological variables explained only a small amount of the overall variance in adherence behaviour, this result provides further support for the hypothesis that psychological variables contribute, in part, to continuous positive airway pressure adherence. Future research should focus on highlighting discrete variables, which may helpfully inform psychologically based interventions aimed at improving the use of continuous positive airway pressure by patients with obstructive sleep apnoea/hypopnoea syndrome at risk of discontinuance. PMID- 15358708 TI - Penetration of newer quinolones in the empyema fluid. AB - The degree of penetration of newer quinolones into the pleural fluid has not been studied. The objective of the present study was to determine the degree to which moxifloxacin and levofloxacin penetrate into empyemic pleural fluid using a new rabbit model of empyema. An empyema was created via the intrapleural injection of turpentine (1 mL), followed 24 h later by instillation of 2 mL (1 x 10(10)) Escherichia coli bacteria (ATCC 35218) into the pleural space of New Zealand white rabbits. After an empyema was verified by thoracentesis and pleural fluid analysis, moxifloxacin and levofloxacin (25 mg.kg(-1) for both, i.v.) were administered. Antibiotic levels were determined in samples of pleural fluid and in blood collected serially over 12 h. Antibiotic levels were measured using HPLC. Each of the antibiotics penetrated well into the empyemic pleural fluid. Antibiotic penetration was the greatest for moxifloxacin (area under the curve (AUC) for pleural fluid/blood (AUCPF/AUCblood) ratio=1.37) followed by levofloxacin (ratio=1.13). The time to equilibration between the pleural fluid and blood antibiotic levels was more rapid for moxifloxacin (3.9 h) than for levofloxacin (4.4 h). With moxifloxacin, the peak pleural fluid concentration (Cmax,PF) was 2.77 microg.mL(-1) and occurred at a time to maximum pleural fluid concentration (Tmax,PF) of 6 h after infusion and decreased thereafter. The peak blood concentration (Cmax,blood) was 4.81 microg.mL(-1) at 1 h after administration. With levofloxacin, the peak pleural fluid level (Cmax,PF=1.39 microg.mL(-1)) occurred at 6 h (Tmax,PF=6 h) after infusion. The Cmax,blood was 1.88 microg.mL(-1) at 1 h after administration. In conclusion, differences were found in the degree of penetration of the two quinolones into infected pleural fluid in rabbits. The clinical significance of these differences is unknown. More studies are needed to evaluate the pharmacokinetic parameters in the pleural space in humans. PMID- 15358709 TI - Post-genome respiratory epidemiology: a multidisciplinary challenge. AB - The introduction of genetic approaches in respiratory epidemiology is novel for most epidemiologists, and the post-genome phase poses new challenges. After describing specific questions pertinent to the field of asthma and chronic obstructive pulmonary disease, two main methodological aspects regarding technological and scientific advances are presented in this review. The first one concerns biological aspects in the genome and post-genome phases, i.e. how to study the genome, the transcriptome and the proteome. The second area concerns genetic epidemiology, considering design (case control and family based) and statistical analytical issues. Key aspects are large sample size, good phenotyping and the consideration of environment-by-gene interaction according to windows of opportunity. Needs that have been identified include the following. 1) Networking for setting standards in the field and access to sufficiently large samples. 2) Multidisciplinarity; the collaboration of epidemiologists, clinicians, geneticists and specialists in bioinformatics, in addition to specialists in disciplines less familiar to epidemiologists, to be prepared for new phenotypic characterisations based on transcriptome and proteome. 3) Training in genetic analytical techniques for some respiratory epidemiologists, as well as in respiratory epidemiology for some genetic epidemiologists. Implications for research, considering ethical aspects, public health aspects and organisational aspects in the field of genetic and environmental respiratory epidemiology also need to be addressed. PMID- 15358710 TI - The diagnosis and management of chronic cough. PMID- 15358711 TI - European framework to decrease the burden of TB/HIV. AB - Tuberculosis (TB) in Europe is declining in countries in western and central Europe, but the burden is still high and increasing in eastern Europe. HIV/AIDS is increasing dramatically in eastern Europe. HIV-related tuberculosis (TB/HIV) morbidity and mortality are expected to accelerate significantly in the future. This framework aims to guide European countries in developing their national plan for reducing TB/HIV morbidity and mortality. It results from an extensive consultation process undertaken by the World Health Organization Regional Office for Europe and by those responsible for HIV/AIDS and TB programmes and their partners. It builds on strategies developed globally and in Europe for TB control and for HIV/AIDS prevention and care. This framework sets out the rationale for effective collaboration between HIV/AIDS and tuberculosis national programmes. It identifies five strategic components (political commitment, collaborative prevention, intensified case-finding, coordinated treatment and strengthened surveillance) and eight key operations (central coordination, policy development, surveillance, training, supply management, service delivery, health promotion and research). PMID- 15358712 TI - Adenocarcinoma of the lung mimicking inflammatory lung disease with honeycombing. AB - Pulmonary adenocarcinoma of the lung and its variants are well-defined entities, since the recent WHO classification of lung tumours. However, scant descriptions have been allocated to associated stromal changes, such as prominent inflammation and fibrosis, which can overshadow a tumoral proliferation and masquerade as a benign reactive process and this has not been recognised as a histopathological variant. The case of a 72-yr-old farmer who presented a multifocal well differentiated adenocarcinoma that mimicked honeycomb lung with bronchiolectasis radiologically, on computed tomography scan and histologically at open lung biopsy, is reported. Histological pitfalls in the biopsy were represented by mild atypical cuboidal or columnar epithelial cells lining bronchiolar structures resembling florid bronchiolar metaplasia in a background of extensive fibrosis and inflammation, features that mimicked inflammatory honeycombing. However, histological analysis of the surgical resection of the main lesion, performed because of a clinical alteration of the patient, confirmed the diagnosis of multifocal adenocarcinoma of mixed subtype. A monomorphic proliferation of clear cells, lack of associated ciliated or squamous cells and presence of significant cytologic atypia gave a diagnosis of malignancy. This case illustrates how inflammatory and fibrotic changes may conceal a correct diagnosis of carcinoma and emphasises the importance of adequate sampling in such cases. PMID- 15358713 TI - A young male with chest pain, cough and fever. PMID- 15358714 TI - Inspiratory muscle training as a tool for the management of patients with COPD. PMID- 15358715 TI - Increased mortality in COPD among construction workers exposed to inorganic dust. PMID- 15358716 TI - Meta-analysis on dose-response relationship of inhaled steroids must be done in homogenous asthma populations. PMID- 15358717 TI - Insulin-sensitizing agents as primary therapy for patients with polycystic ovarian syndrome. AB - BACKGROUND: This paper is a systematic review of metformin versus clomiphene citrate (CC) in women with polycystic ovary syndrome (PCOS). METHODS: Meta analysis Of Observational Studies in Epidemiology (MOOSE) and QUality Of Reporting Of Meta-analyses (QUOROM) guidelines were followed. A systematic computerized literature search was done of seven bibliographic databases. Inclusion criteria included cohort and randomized controlled trials (RCT) of women with PCOS and the following medications: metformin versus placebo; metformin versus CC; metformin plus CC versus placebo plus CC. Rev-man 4.1 and Metaview 4.0 were used to analyse data. Relative risk (RR) estimates were presented. A chi2-test determined the significance of the association. Heterogeneity was determined by the Cochran Q-test. RESULTS: Metformin was 50% better than placebo for ovulation induction in infertile PCOS patients [RR 1.50; 95% confidence interval (CI) 1.13, 1.99]. Metformin was also of benefit in non infertile (i.e. patients with PCOS who were not complaining of infertility) PCOS patients for cycle regulation compared to placebo (RR 1.45; CI 1.11, 1.90). Metformin was not of confirmed benefit versus placebo for achievement of pregnancy (RR 1.07; CI 0.20, 5.74). Metformin plus CC may be 3-4-fold superior to CC alone for ovulation induction (RR 3.04; CI 1.77, 5.24) and pregnancy (RR 3.65; CI 1.11, 11.99) in women with PCOS. CONCLUSIONS: Metformin is effective for ovulation induction and cycle regulation in this group of patients. Metformin plus CC appears to be very effective for achievement of pregnancy compared to CC alone. No RCTs directly compare metformin to CC but the need for such a trial exists. PMID- 15358718 TI - Effect of pituitary downregulation on antral follicle count, ovarian volume and stromal blood flow measured by three-dimensional ultrasound with power Doppler prior to ovarian stimulation. AB - BACKGROUND: Despite the extensive use of gonadotrophin releasing hormone agonists (GnRH ag) for pituitary downregulation, the literature regarding their effect on ultrasound parameters for predicting ovarian responses are few and conflicting. The aim of this prospective study was to compare antral follicle count (AFC), ovarian volume and ovarian stromal blood flow measured by three-dimensional (3D) power Doppler ultrasound before and after pituitary downregulation. METHODS: All patients received a long protocol of intranasal Buserelin from the mid-luteal phase of the cycle. In the early follicular phase of the preceding cycle before downregulation and on the second day of the treatment cycle after downregulation, patients underwent a blood test for serum FSH, LH and estradiol and a transvaginal scanning with 3D power Doppler to determine AFC, ovarian volume and ovarian 3D power Doppler flow indices. RESULTS: Out of 104 women scanned, 85 women were analysed. Polycystic ovaries were encountered in 14 (16.5%) women. No significant differences in any of the ultrasound parameters were demonstrated before and after downregulation, in patients with normal ovaries or in those with polycystic ovaries. CONCLUSION: AFC, ovarian volume and ovarian 3D power Doppler flow indices did not significantly change after a short-term treatment of GnRH agonist for pituitary downregulation. PMID- 15358719 TI - What is the most relevant standard of success in assisted reproduction?: The cumulated singleton/twin delivery rates per oocyte pick-up: the CUSIDERA and CUTWIDERA. AB - National and international registries are essential tools for establishing new standards and comparing success rates, but they do not take into account the total pregnancy/delivery rate per oocyte recovery. In Switzerland and Germany, because of legal constraints, a maximum of three two-pronuclear zygotes are allocated for transfer whereas all the supernumerary pronuclear zygotes are immediately cryopreserved, preventing selection of the transferred embryos. We report on a 10 years' experience (1993-2002) of our centre which performs transfers of unselected embryos and cryopreservation at the two-pronuclear zygote stage. As approximately 30% of all deliveries are from cryo cycles, it is essential to take into account the contribution of the cryo transfers, and we propose therefore to evaluate, as a measure of IVF performance, the cumulated delivery rate per oocyte pick-up. This delivery rate is broken down further into the cumulated singleton delivery rate (CUSIDERA) and the cumulated twin delivery rate (CUTWIDERA). The sum (S) of these two rates is a measure of efficacy while the ratio CUTWIDERA/S as a percentage is a measure of safety of IVF treatments. Using these new indexes, the average 10 year efficacy and safety of our IVF programme were 26 and 19%, respectively. Both CUSIDERA and CUTWIDERA can be calculated easily in any clinical situation and yield useful parameters for patient counselling and internal/external benchmarking purposes. PMID- 15358720 TI - Semen parameters of a semen donor before and after infection with human immunodeficiency virus type 1: case report. AB - Semen samples from a donor who seroconverted for human immunodeficiency virus type 1 (HIV-1) during the period that he was donating at our clinic were stored before and after infection. Semen analysis was done on all of these samples before cryopreservation. Retrospectively, both qualitative and quantitative HIV-1 testing was performed on the cryopreserved semen samples to determine the time of primary HIV-1 infection. After HIV-1 infection, semen volume, sperm motility and the percentage of spermatozoa with normal morphology were reduced compared with the same parameters before HIV-1 infection. HIV-1 RNA was intermittently detectable in semen. HIV-1 infection led to a reduction in semen volume, sperm motility and normal sperm morphology in this donor. However, the clinical significance of these findings is unclear. A longitudinal cohort study on the effects of HIV-1 infection on semen quality is necessary to confirm these findings. PMID- 15358721 TI - Psychological follow-up study of 5-year-old ICSI children. AB - BACKGROUND: The developmental outcomes of children born after ICSI are still a matter of concern. The purpose of the present study was to investigate psychological outcomes for 5-year-old children born after ICSI and compare these with outcomes for children born after spontaneous conception (SC). METHODS: Three hundred singleton children born after ICSI in Belgium, Sweden and the USA were matched by maternal age, child age and gender. Outcome measures included the Wechsler Preschool and Primary scales of intelligence (WPPSI-R), Peabody Developmental Motor Scales, Parenting Stress Index and Child Behaviour Checklist. RESULTS: Regarding cognitive development, no significant differences were found on WPPSI-R verbal and performance scales between ICSI and SC children. However, some differences were noted on subtests of the Performance Scale. ICSI children more often obtained a score below 1 SD of the mean on the subtests: Object Assembly, Block Design and Mazes (all P<0.05). Significant differences by site (i.e. Belgium, Sweden and New York) were found on subtests related to parenting stress, child behaviour problems and motor development (all P<0.05). These findings can probably be explained by variables other than conception mode, such as cultural differences and selection bias. CONCLUSIONS: Although the finding that a higher proportion of ICSI children obtained scores below the cut-off on some of the visual-spatial subscales of the WPPSI-R warrants further investigation, ICSI does not appear to affect the psychological well-being or cognitive development at age 5. PMID- 15358722 TI - Patterns of hormonal response to the GnRH agonist leuprolide in brothers of women with polycystic ovary syndrome: a pilot study. AB - BACKGROUND: The aim of this study was to evaluate the effect of a single dose of leuprolide acetate on gonadotrophin and gonadal steroid secretion in brothers of women with polycystic ovary syndrome (PCOS), in order to assess P450c17alpha activity. An oral glucose tolerance test (OGTT) and a lipid profile were also performed. METHODS: Twenty-two unrelated brothers of women with PCOS (PCOS(b)) and 14 brothers of normal cycling women (C(b)), matched for age, underwent a leuprolide acetate test (10 microg/kg s.c.) and an OGTT with measurement of circulating concentrations of gonadotrophins, steroid hormones, glucose, insulin and lipids. RESULTS: Clinical and basal hormonal parameters were similar in both groups. After leuprolide administration, PCOS(b) exhibited a significant increase of 17alpha-hydroxyprogesterone (17-OHP) compared to C(b) (P<0.05). However, only 45% of PCOS(b) showed a supranormal increase of 17-OHP (2 SD above the respective control group mean values, P<0.003) with a normal gonadotrophin response (group 1). The other 55% of the PCOS(b) exhibited a normal 17-OHP response to the analogue (group 2). However, in group 2, basal steroid concentrations did not show a uniform pattern: six of the PCOS(b) exhibited high basal androstenedione (2 SD above the respective control group mean values), three were very similar to C(b), and the other three presented lower basal testosterone concentrations (2 SD below the respective control group mean values) than those observed in C(b). CONCLUSIONS: This study shows that different responses to leuprolide in PCOS brothers make evident the heterogeneity of this syndrome in which P450c17alpha activity could be involved. PMID- 15358723 TI - An update on the genetics of colorectal cancer. AB - Colorectal carcinoma (CRC) remains a frequent cause of cancer-associated mortality in the UK and still has a relatively poor outcome. Single gene defects account for up to 2-6% of cases, but twin studies suggest a hereditary component in 35%. CRC represents a paradigm for cancer genetics. Almost all the major-gene influences on CRC have been identified, and the identification of the remaining susceptibility alleles is proving troublesome. Only a few low-penetrance alleles, such as methylene tetrahydrofolate reductase C677T, appear convincingly to be associated with CRC risk. To identify the remaining CRC genes, parallel approaches, including strategies based on linkage and association and complementary analyses such as searches for modifier genes, must be employed. To gain sufficient evidence to prove that a gene is involved in CRC predisposition, it is probably necessary for multiple, adequately-powered studies to demonstrate an association with the disease, especially if the allelic variants have only a small differential effect on risk. It may also be possible to show how genes interact with each other and the environment, although this will be even more difficult. Accurate quantitation of the allele-specific risks in different populations will be necessary, but problematic, especially if those risks combine in a fashion which is not of a straightforward additive or multiplicative type. Without any good prior evidence of the nature of the remaining genetic influence on CRC, the possibility remains that this is a truly polygenic trait or that multiple, rare variants contribute to the increased risk; in these cases, identification of the genes involved will be very difficult. Despite these potential problems, the effectiveness of preventive measures for CRC, especially in high-risk individuals, means that the search for new predisposition genes is justified. PMID- 15358724 TI - Genetics of infectious diseases. AB - Infectious diseases represent a major health problem worldwide, both in terms of morbidity and mortality. A complex combination of environmental, pathogen and host genetic factors plays a role in determining both susceptibility to particular microbes and the course of infection. Numerous studies have now mapped and identified relevant genes using a variety of both family-based and population based approaches. Much interest has been focused on susceptibility to malaria, HIV/AIDS and mycobacterial infection, but other bacterial, viral and parasitic diseases are receiving increasing attention. Some major genes have been identified by genome scans of multi-case families, and mouse genetics has contributed to mapping and identification of a few genes. However, the great majority of known susceptibility loci emerged from screening of likely candidate genes. The emerging picture is of highly polygenic diseases, with occasional major genes, along with significant inter-population heterogeneity. This genetic architecture likely reflects the role that evolutionary selection has played in generating and maintaining a diverse repertoire of susceptibility/resistance loci, most with individually small effects. Genome-wide association studies with large sample sizes will be required to define the majority of the relevant polygenes. PMID- 15358725 TI - Molecular genetics of distal hereditary motor neuropathies. AB - Inherited peripheral neuropathies comprise a wide variety of diseases primarily affecting the peripheral nervous system. The best-known peripheral neuropathy is Charcot-Marie-Tooth disease (CMT) described in 1886 by J.-M. Charcot, P. Marie and H.H. Tooth. In 1980, A.E. Harding and P.K. Thomas showed that in a large group of individuals with CMT, several only had motor abnormalities on clinical and electrophysiological examination, whereas sensory abnormalities were absent. This exclusively motor variant of CMT was designated as spinal CMT or hereditary distal spinal muscular atrophy, and included in the distal hereditary motor neuropathies (distal HMN). The distal HMN are clinically and genetically heterogeneous and are subdivided according to the mode of inheritance, age at onset and clinical evolution. Since the introduction of positional cloning, 12 chromosomal loci and seven disease-causing genes have been identified for autosomal dominant and recessive distal HMN. Most of the genes involved have housekeeping functions, as in RNA processing, translation synthesis, glycosylation, stress response, apoptosis, but also axonal trafficking and editing. Functional characterization of the mutations will help to unravel the cellular processes that underlie the specificity of motor neuropathies leading to neurogenic muscular atrophy of distal limb muscles. Here we review the recent progress of the molecular genetics of distal HMN and discuss the genes implicated. PMID- 15358726 TI - Thalassemia in Sri Lanka: a progress report. AB - The thalassemias pose an increasing burden for health-care services in many Asian countries. In order to conserve rare resources, it is essential to determine the reasons for the remarkable phenotypic heterogeneity and natural history of these disorders so that the most cost-effective methods for their control and management can be established. A long-term observational study of patients with different forms of thalassemia in Sri Lanka suggests that in addition to the well defined primary, secondary and tertiary genetic modifiers, environmental factors, particularly malaria, and variation in the ability to adapt to the profound anaemia which characterizes these conditions, may play a significant role in determining their clinical severity. These findings may have important implications for the control and management of thalassemia in Asian populations. PMID- 15358727 TI - Genetic disorders influencing lung formation and function at birth. AB - Adaptation to air breathing at birth is dependent on formation and function of the lung. Lung morphogenesis is a complex process dependent on precise temporal spatial control of cell proliferation, differentiation and behavior mediated by autocrine-paracrine signaling that instructs transcriptional processes during organogenesis. Mutations in genes causing severe, and often lethal, lung malformations include those in the sonic hedgehog, fibroblast growth factor and thyroid transcription factor-1 pathways. Mutations in genes regulating surfactant homeostasis, necessary for reduction of surface tension in the alveoli, cause lethal respiratory distress at birth or interstitial lung disease in childhood. Inherited disorders of the surfactant system that affect neonatal respiratory adaptation at birth include hereditary surfactant protein B deficiency, mutations in surfactant protein C and the ABCA3 transporter. PMID- 15358728 TI - Environmental genomics: a key to understanding biology, pathophysiology and disease. AB - Recent advances in human and molecular genetics provide an unparalleled opportunity to understand how genes and genetic changes interact with environmental stimuli to either preserve health or cause disease. The fields of environmental genetics and environmental genomics has enormous potential to affect our ability to accurately assess the risk of developing disease, identify and understand basic pathogenic mechanisms that are critical to disease progression, and to more precisely phenotype disease subtypes. However, the application of genetics and genomics to problems in environmental health is only the beginning yet, by itself, represents a potentially effective strategy to substantially impact morbidity and mortality. Collaborative approaches that team together environmental scientists with molecular biologists, geneticists, physiologists and physician scientists are critical to the investigation of environmental aspects of human health. Moreover, exploiting eukaryotic model systems (yeast, Caenorhabditis elegans, zebrafish, Drosophila and rodents) will accelerate our understanding of environmental exposures on human health. PMID- 15358729 TI - Mechanisms of non-Mendelian inheritance in genetic disease. AB - Single gene disorders with Mendelian inheritance patterns have contributed greatly to the identification of genes and pathways implicated in genetic disease. In these cases, molecular analysis predicts disease status relatively directly. However, there are many abnormalities which show familial recurrence and have a clear genetic component, but do not show regular Mendelian segregation patterns. Defining the causative gene for non-Mendelian diseases is more difficult, and even when the underlying gene is known, there is uncertainty for prenatal prediction. However, detailed examination of the different mechanisms that underlie non-Mendelian segregation provides insight into the types of interaction that regulate more complex disease genetics. PMID- 15358730 TI - Polyalanine expansions in human. AB - Beside the well-known polyglutamine expansions involved in several neurodegenerative disorders, convergent recent findings pointed to the expansion of polyalanine stretches as a disease mechanism in congenital malformations, skeletal dysplasia and nervous system anomalies. Polyalanine stretches have been predicted in roughly 500 human proteins among which nine have been ascribed to disease phenotype by expansion of polyalanines. The function of polyalanine stretches is largely unknown. This paper aims to review the rapidly growing evidences for a disease-causing mechanism common to expansion of homopolymeric tracts whatever the amino acid involved is. PMID- 15358731 TI - Positive selection on the human genome. AB - Positive selection has undoubtedly played a critical role in the evolution of Homo sapiens. Of the many phenotypic traits that define our species--notably the enormous brain, advanced cognitive abilities, complex vocal organs, bipedalism and opposable thumbs--most (if not all) are likely the product of strong positive selection. Many other aspects of human biology not necessarily related to the 'branding' of our species, such as host-pathogen interactions, reproduction, dietary adaptation and physical appearance, have also been the substrate of varying levels of positive selection. Comparative genetics/genomics studies in recent years have uncovered a growing list of genes that might have experienced positive selection during the evolution of human and/or primates. These genes offer valuable inroads into understanding the biological processes specific to humans, and the evolutionary forces that gave rise to them. Here, we present a comprehensive review of these genes, and their implications for human evolution. PMID- 15358732 TI - Allele-specific gene expression differences in humans. AB - In the last decade, the search for the genetic origins of phenotypic variation has expanded beyond the non-synonymous variants which alter the amino acid sequence of the encoded protein, and many examples of sequence variants which alter gene expression have been found. Recently, using both traditional and novel technologies, a number of surveys have been carried out to examine the frequency with which cis-acting sequence variants or other cis-acting effects, alter gene expression either in vitro or in vivo. Microarray data have shown that the expression of many genes varies markedly between individuals and allele-specific expression studies have shown that the source of much of this variation appears to be cis-acting effects. A significant proportion of the variation may originate in gene promoter regions and a large number of sequence variants which have functional effect in vitro have been found. The evidence suggests that given a large enough population, most, if not all genes may have allele-specific expression differences in at least some individuals and finding the genetic origins of each of these and linking the former to a possible phenotype must be a major long term goal of the biomedical community. PMID- 15358733 TI - Exploiting human--fish genome comparisons for deciphering gene regulation. AB - Comparative genomics has served as an essential guide in the identification of functional coding and non-coding sequences in vertebrate genomes. Human-mouse pair-wise comparisons have limited utility for identifying functional conserved non-coding sequences, owing to the large number of sequences shared between these species. In searching for more stringent filters to uncover non-coding elements more likely to be of functional importance in the human genome, human-fish sequence comparisons have emerged as an important strategy, leading to the efficient identification of enhancer elements. These sequences are unevenly distributed in the genome, tending to cluster around genes involved in key developmental processes, with recent studies suggesting that they represent genomic segments in which sequence variation can result in morphological changes and innovation. These elements, conserved over long evolutionary time, emerge as primary candidates that are likely to harbor sequence variation contributing to susceptibility of human disease phenotypes. PMID- 15358734 TI - 'Am not I a fly like thee?' From genes in fruit flies to behavior in humans. AB - The fruit fly Drosophila melanogaster has provided insight into the role of genes in behavior, some of which have relevant implications for humans. Mutants induced or engineered in the laboratory have contributed to our understanding of biological rhythms, learning, memory, neurodegenerative disease and drug response. Studies of naturally occurring genetic variation in behavior have advanced our understanding of what kinds of variants arise spontaneously and contribute to behavior. PMID- 15358735 TI - RNA interference: from model organisms towards therapy for neural and neuromuscular disorders. AB - Experimental RNA interference (RNAi) leading to the selective knockdown of gene function is induced by introducing into cells either double stranded RNA (dsRNA), or short interfering RNA (siRNA) fragments into which dsRNA is cut. The siRNA triggers degradation of homologous messenger RNA (mRNA). Widely used as a research tool in the genetic model organisms Caenorhabditis elegans, Drosophila melanogaster and mouse to investigate the function of individual genes, RNAi has also been deployed in genome-wide, specific gene-knockdown screens. Recent rapid progress in the application of RNAi to mammalian cells, including neurons and muscle cells, offers new approaches to drug target identification and validation. Advances in targeted delivery of RNAi-inducing molecules has raised the possibility of using RNAi directly as a therapy for a variety of human genetic and other neural and neuromuscular disorders. Here, we review examples of the application of RNAi to worm, fly and mouse models of such diseases aimed at understanding their pathophysiology and we address problems to be solved in developing RNAi-based therapies. PMID- 15358736 TI - The developmental genetics of auditory hair cells. AB - Loss of auditory hair cells (AHCs) is a major cause of human deafness. Considerable effort has been devoted to unraveling how these mechanotransducers of sound are specified, with a view to correcting hearing loss by gene or stem cell therapies. Recent work on signaling cascades, particularly lateral inhibition and planar cell polarity, has begun to tie together some of the known pathways. Mutations in mice and humans that cause hearing and/or balance disorders are also shedding light on how AHCs are specified and, maintained and handle ion flux. Studies on some of these genes are beginning to provide insights into the more complex genetics of later onset forms of hearing loss. Progress has also been made in solving some long-term goals of auditory biology. Cadherin23 has been identified as a component of AHC stereocilia tip links, and progress has been made towards identifying the elusive AHC mechanoreceptor channel. Preliminary steps have also been taken towards inner-ear gene therapy, and in the engineering of embryonic stem cells for eventual cell therapies. Mammals cannot regenerate AHCs, but birds and other lower vertebrates can. Genomic tools have now been brought to bear on this problem with the aim of deciphering the molecular basis of this regenerative capability. The combination of new genomic tools and the many mouse and chicken embryological and genetic resources should increasingly provide new insights into how AHCs are programed and maintained. PMID- 15358737 TI - Applications of genomic microarrays to explore human chromosome structure and function. AB - The combination of genomic microarrays with comparative genomic hybridization and with chromatin immunoprecipitation is providing an increasingly detailed view of the way in which the human genome is organized and functions and how disorganization and disfunction can lead to disease. These studies are enhanced by the flexibility of array technology, allowing resolutions from coverage of the whole genome using 200 kb cloned DNA inserts to detailed analysis using PCR products or oligonucleotides of 100 bp or less. In particular, the use of chromatin immunoprecipitation is providing new insights into chromosome structure and gene regulation and control through the analysis of protein--DNA interactions. PMID- 15358738 TI - Human chromosome 7 circa 2004: a model for structural and functional studies of the human genome. AB - Human chromosome 7 is arguably amongst the most comprehensively characterized segments of the human genome. By microscopic examination, it belongs to the medium-sized group C submetacentric class, and historical studies involving chromosome-length measurements estimated that it accounts for approximately 5.3% of the human genome (or 160 Mb). Early successes in molecular genetics led to the identification of some of its biomedically important genes, including the T-cell receptor and homeobox families as well as the erythropoietin and cystic fibrosis genes. The Human Genome Project brought chromosome 7-specific and genome-wide initiatives, generating a wealth of genomic resources that have revealed the presence of over 350 disease-associated genes. Two distinct assemblies of the chromosome 7 sequence have been generated-one based largely on mapped large insert clones and the other based on an integrated whole-genome shotgun sequencing strategy. These two sequences are mainly identical (<1% difference), and both estimate the unit length of chromosome 7 to be just over 158 Mb, remarkably similar to the originally predicted size. Systematic annotation efforts have anchored to the sequence, amongst many features, over 900 known genes and some 1000 other gene structures, as well as over 650 chromosomal breakpoints identified in patients with characterized phenotypic differences. Chromosome 7 has also been shown to contain the highest content of intra autosomal segmental duplications in the human genome. The orthologous regions of roughly 22 Mb of chromosome 7 are currently being sequenced in multiple other vertebrate species. Examining these comparative sequence data, in conjunction with the other accumulating genomic information about these regions and the rest of the chromosome, should provide a model for the next generation of structural and functional analyses of the human genome. PMID- 15358739 TI - Prediction of bone mineral density from vitamin D receptor polymorphisms is uncertain in representative samples of Japanese Women. The Japanese Population based Osteoporosis (JPOS) Study. AB - BACKGROUND: Vitamin D receptor (VDR) gene polymorphisms have been inconsistently associated with bone mineral density (BMD). To precisely evaluate the associations between three VDR gene polymorphisms and BMD, we performed a large scale representative study of the Japanese female population. METHODS: Fifty women were randomly selected from each of the 5-year age stratified populations (15-79 years) in each of the three municipalities examined, as a part of the Japanese population-based osteoporosis (JPOS) baseline study in 1996. In the study, BMD at the lumbar spine, hip, and distal forearm were measured using dual energy X-ray absorptiometry (DXA). Two polymorphisms were determined in the VDR gene locus identified by the restriction endonucleases ApaI and TaqI through a novel allele discrimination method using two different allele-specific fluorescence probes. The other VDR gene polymorphism was identified by the restriction endonuclease FokI using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Changes in BMD were determined in a follow-up study 3 years after the baseline study. RESULTS: After the exclusion of women who had any medical or menstrual history affecting BMD, 1434 women were analysed. The annual per cent changes in BMD at the lumbar spine over 3 years in subjects with tt genotype in the TaqI polymorphism were different from other genotypes, both in the women who were premenopausal at the follow-up survey (F premenopausal women) and in the women who were postmenopausal at the baseline survey (B-postmenopausal women). However, the effects of tt genotype on change in BMD were opposite in the two groups. In addition, these associations or tendencies were not observed at the different skeletal sites. CONCLUSIONS: This study revealed that none of the individual VDR gene polymorphisms displayed consistent association with baseline BMD or BMD change. Therefore, the effect of the VDR genotype on bone mass is negligible in Japanese women. PMID- 15358740 TI - Does the misreporting of adult body size depend upon an individual's height and weight? Methodological debate. PMID- 15358741 TI - Mathematical coupling: a multilevel approach. PMID- 15358742 TI - Assessing correlation between reporting errors and true values: untestable assumptions are unavoidable. PMID- 15358743 TI - Mathematical coupling: a simpler approach. PMID- 15358745 TI - Further development of the case-only design for assessing gene-environment interaction: evaluation of and adjustment for bias. AB - BACKGROUND: The case-only study for investigating gene-environment interactions provides increased statistical efficiency over case-control analyses. This design has been criticized for being susceptible to bias arising from non-independence between the genetic and environmental factors in the population. Given that independence is critical to the validity of case-only estimates of interaction, researchers frequently use controls to evaluate whether the independence assumption is tenable, as advised in the literature. Our work investigates to what extent this approach is appropriate and how non-independence can be accounted for in case-only analyses. METHODS: We provide a formula in epidemiological terms that illustrates the relationship between the gene environment association measured among controls and the gene-environment association in the source population. Using this formula, we conducted sensitivity analyses to describe the circumstances in which controls can be used as proxy for the source population when evaluating gene-environment independence. Lastly, we generated hypothetical cohort data to examine whether multivariable modelling approaches can be used to control for non-independence. RESULTS: Our sensitivity analyses show that controls should not be used to evaluate gene environment independence in the population, even when the baseline risk of disease is low (i.e. 1%), and the interaction and independent effects are moderate (i.e. risk ratio = 2). When the factors are associated, it is possible to remove bias arising from non-independence using standard statistical multivariable techniques in case-only analyses. CONCLUSIONS: Even when the disease risk is low, evaluation of gene-environment independence in controls does not provide a consistent test for bias in the case-only study. Given that control for non-independence is possible when the source of the non-independence can be conceptualized, the case-only design may still be a useful epidemiological tool for examining gene-environment interactions. PMID- 15358746 TI - Influences on childhood height: comparing two generations in the 1958 British birth cohort. AB - BACKGROUND: Genetics and early environment are known to influence height, but evidence is sparse on changes in these influences over time. METHODS: The 1958 British birth cohort study includes all children born between March 3, 1958 and March 9, 1958, who were followed to age 41 yr, and one-third of their offspring in 1991. Childhood height in each generation (measured at 7 yr for cohort members and 4-18 yr for offspring) was converted to a standard deviation score based on the 1990 British growth reference. We used multilevel models to analyse influences on height in order to allow for the hierarchical within-family data structure. RESULTS: Childhood height increased by 1 cm between 1958 cohort members and their offspring. Several influences on childhood height in the older generation (maternal smoking, breastfeeding, maternal age, social class, maternal education, and parental divorce) did not affect childhood height in the younger generation. Parental height was most strongly associated with childhood height and effects did not diminish between generations [adjusted increase approximately 2 cm for 1 maternal or paternal height standard deviation score (SDS)]. Third- or later-borns and those with three or more siblings had deficits of 1-2 cm (adjusted estimates) in both generations. Other factors, particularly indicators of socioeconomic position, showed weaker effects in the younger generation. For example, the growth deficit of 1.1 cm (adjusted estimate) among cohort members from households with >1.5 persons/room had disappeared in the offspring. CONCLUSIONS: Within Great Britain, the adverse effects of environmental factors on childhood height have lessened between recent generations. PMID- 15358747 TI - The New Epidemiology: putting the pieces together in complex disease aetiology. PMID- 15358749 TI - Use of intramuscular triacylglycerol as a substrate source during exercise in humans. AB - Fat and carbohydrate are the principal substrates that fuel aerobic ATP synthesis in skeletal muscle. Most endogenous fat is stored as triacylglycerol in subcutaneous and deep visceral adipose tissue. Smaller quantities of triacylglycerol are deposited as lipid droplets inside skeletal muscle fibers. The potential role of intramyocellular triacylglycerol (IMTG) as a substrate source during exercise in humans has recently regained much of its interest because of the proposed functional relationship between IMTG accumulation and the development of skeletal muscle insulin resistance. Exercise likely represents an effective means to prevent excess IMTG accretion by stimulating its rate of oxidation. However, there is much controversy on the actual contribution of the IMTG pool as a substrate source during exercise. The apparent discrepancy in the literature likely stems from methodological difficulties that have been associated with the methods used to estimate IMTG oxidation during exercise. However, recent studies using stable isotope methodology, 1H-magnetic resonance spectroscopy, and electron and/or immunofluorescence microscopy all support the contention that the IMTG pool can function as an important substrate source during exercise. Although more research is warranted, IMTG mobilization and/or oxidation during exercise seem to be largely determined by exercise intensity, exercise duration, macronutrient composition of the diet, training status, gender, and/or age. In addition, indirect evidence suggests that the capacity to mobilize and/or oxidize IMTG is substantially impaired in an obese and/or Type 2 diabetic state. As we now become aware that skeletal muscle has an enormous capacity to oxidize IMTG stores during exercise, more research is warranted to develop combined exercise, nutritional, and/or pharmacological interventions to effectively stimulate IMTG oxidation in sedentary, obese, and/or Type 2 diabetes patients. PMID- 15358750 TI - Doxycycline effects on mechanical and morphometrical properties of early- and late-stage osteoarthritic bone following anterior cruciate ligament injury. AB - As posttraumatic osteoarthritis (OA) progresses, the mechanical and morphometrical properties of the subchondral bone change and may be linked to damage of the articular cartilage. Potentially to slow that progression, doxycycline was administered orally twice daily (4 mg.kg(-1).day(-1)) in skeletally mature canines after anterior cruciate ligament transection (ACLX). To test if doxycycline significantly altered the structure and function of OA bone, we tested cancellous bone mechanical properties, measured bone mineral content, and analyzed bone structure by microcomputed tomography. Our investigation focused on subchondral trabecular bone changes in the medial femoral condyle at 36 and 72 wk after ACLX. Significant mechanical changes discovered at 36 wk post ACLX were less obvious at 72 wk in both treated and ACLX groups. Doxycycline treatment conserved bone strain energy density at 72 wk. Doxycycline had little effect on the degradation of superficial osseous tissue at 36 wk post-ACLX; by 72 wk, doxycycline in an ACLX model limited subchondral bone loss within the first 3 mm of periarticular bone with established OA. Significant bone loss occurred in the deeper trabecular bone for all groups. Substantial architectural adaptation within deeper trabecular bone accompanied changes in mechanics in early and established OA. PMID- 15358751 TI - Branched-chain amino acid supplementation and human performance when hypohydrated in the heat. AB - The serotonin system may contribute to reduced human performance when hypohydrated in the heat. This study determined whether branched-chain amino acid (BCAA) supplementation could sustain exercise and cognitive performance in the heat (40 degrees C dry bulb, 20% relative humidity) when hypohydrated by 4% of body mass. Seven heat-acclimated men completed two experimental trials, each consisting of one preparation and one test day. On day 1, a low-carbohydrate diet was eaten and subjects performed exhaustive cycling (morning) and treadmill exercise in the heat (afternoon) to lower muscle glycogen and achieve the desired hypohydration level. On day 2, subjects consumed an isocaloric BCAA and carbohydrate (BC) or carbohydrate-only drink during exercise. Experimental trials included 60 min of cycle ergometry (50% peak oxygen uptake) followed by a 30-min time trial in the heat. A cognitive test battery was completed before and after exercise, and blood samples were taken. BC produced a 2.5-fold increase (P < 0.05) in plasma BCAA and lowered (P < 0.05) the ratios of total tryptophan to BCAA and large neutral amino acid. Blood prolactin, glucose, lactate, and osmolality were not different between trials but increased over time. Cardiovascular and thermoregulatory data were also similar between trials. BC did not alter time-trial performance, cognitive performance, mood, perceived exertion, or perceived thermal comfort. We conclude that BCAA does not alter exercise or cognitive performance in the heat when subjects are hypohydrated. PMID- 15358752 TI - Lung volumes and alveolar expansion pattern in immature rabbits treated with serum-diluted surfactant. AB - In acute respiratory distress syndrome, mechanical ventilation often induces alveolar overdistension aggravating the primary insult. To examine the mechanism of overdistension, surfactant-deficient immature rabbits were anesthetized with pentobarbital sodium, and their lungs were treated with serum-diluted modified natural surfactant (porcine lung extract; 2 mg/ml, 10 ml/kg). By mechanical ventilation with a peak inspiration pressure of 22.5 cm H2O, the animals had a tidal volume of 14.7 ml/kg (mean), when 2.5 cm H2O positive end-expiratory pressure was added. This volume was similar to that in animals treated with nondiluted modified natural surfactant (24 mg/ml in Ringer solution, 10 ml/kg). However, the lungs fixed at 10 cm H2O on the deflation limbs of the pressure volume curve had the largest alveolar/alveolar duct profiles (> or =48,000 microm2), accounting for 38% of the terminal air spaces, and the smallest (<6,000 microm2), accounting for 31%. These values were higher than those in animals treated with nondiluted modified natural surfactant (P <0.05). We conclude that administration of serum-diluted surfactant to immature neonatal lungs leads to patchy overdistension of terminal air spaces, similar to the expansion pattern that may be seen after dilution of endogenous surfactant with proteinaceous edema fluid in acute respiratory distress syndrome. PMID- 15358753 TI - Lactate dehydrogenase expression at the onset of altered loading in rat soleus muscle. AB - Both functional overload and hindlimb disuse induce significant energy-dependent remodeling of skeletal muscle. Lactate dehydrogenase (LDH), an important enzyme involved in anaerobic glycolysis, catalyzes the interconversion of lactate and pyruvate critical for meeting rapid high-energy demands. The purpose of this study was to determine rat soleus LDH-A and -B isoform expression, mRNA abundance, and enzymatic activity at the onset of increased or decreased loading in the rat soleus muscle. The soleus muscles from male Sprague-Dawley rats were functionally overloaded for up to 3 days by a modified synergist ablation or subjected to disuse by hindlimb suspension for 3 days. LDH mRNA concentration was determined by Northern blotting, LDH protein isoenzyme composition was determined by zymogram analysis, and LDH enzymatic activity was determined spectrophotometrically. LDH-A mRNA abundance increased by 372%, and LDH-B mRNA abundance decreased by 43 and 31% after 24 h and 3 days of functional overload, respectively, compared with that in control rats. LDH protein expression demonstrated a shift by decreasing LDH-B isoforms and increasing LDH-A isoforms. LDH-B activity decreased 80% after 3 days of functional overload. Additionally, LDH-A activity increased by 234% following 3 days of hindlimb suspension. However, neither LDH-A or LDH-B mRNA abundance was affected following 3 days of hindlimb suspension. In summary, the onset of altered loading induced a differential expression of LDH-A and -B in the rat soleus muscle, favoring rapid energy production. Long-term altered loading is associated with myofiber conversion; however, the rapid changes in LDH at the onset of altered loading may be involved in other physiological processes. PMID- 15358754 TI - Cannabinoid receptor expression in peripheral arterial chemoreceptors during postnatal development. AB - Prenatal exposure to tobacco smoke increases risk of sudden infant death syndrome (SIDS). Marijuana is frequently smoked in conjunction with tobacco, and perinatal exposure to marijuana is associated with increased incidence of SIDS. Abnormalities in peripheral arterial chemoreceptor responses during sleep may be operative in infants at risk for SIDS, and nicotine exposure adversely affects peripheral arterial chemoreceptor responses. To determine whether marijuana could potentially affect the activity of peripheral arterial chemoreceptors during early postnatal development, we used in situ hybridization histochemistry to characterize the pattern and level of mRNA expression for cannabinoid type 1 receptor (CB1R) in the carotid body, superior cervical ganglia (SCG), and nodose petrosal-jugular ganglia (NG-PG-JG) complex in newborn rats. We used immunohistochemistry and light, confocal, and electron microscopy to characterize the pattern of CB1R and tyrosine hydroxylase protein expression. CB1R mRNA expression was intense in the NG-PG-JG complex, low to moderate in the SCG, and sparse in the carotid body. With maturation, CB1R gene expression significantly increased (P < 0.01) in the NG-PG-JG complex. CB1R immunoreactivity was localized to nuclei of ganglion cells in the SCG and NG-PG-JG complex, whereas tyrosine hydroxylase immunoreactivity was localized to the cytoplasm. Exposure to marijuana during early development could potentially modify cardiorespiratory responses via peripheral arterial chemoreceptors. The novel finding of nuclear localization of CB1Rs in peripheral ganglion cells suggests that these receptors may have an, as yet, undetermined role in nuclear signaling in sensory and autonomic neurons. PMID- 15358755 TI - Chemical profile of vagal preganglionic motor cells innervating the airways in ferrets: the absence of noncholinergic neurons. AB - In ferrets, we investigated the presence of choline acetyltransferase (ChAT), vasoactive intestinal peptide (VIP), and markers for nitric oxide synthase (NOS) in preganglionic parasympathetic neurons innervating extrathoracic trachea and intrapulmonary airways. Cholera toxin beta-subunit, a retrograde axonal transganglionic tracer, was used to identify airway-related vagal preganglionic neurons. Double-labeling immunohistochemistry and confocal microscopy were employed to characterize the chemical nature of identified airway-related vagal preganglionic neurons at a single cell level. Physiological experiments were performed to determine whether activation of the VIP and ChAT coexpressing vagal preganglionic neurons plays a role in relaxation of precontracted airway smooth muscle tone after muscarinic receptor blockade. The results showed that 1) all identified vagal preganglionic neurons innervating extrathoracic and intrapulmonary airways are acetylcholine-producing cells, 2) cholinergic neurons innervating the airways coexpress ChAT and VIP but do not contain NOS, and 3) chemical stimulation of the rostral nucleus ambiguus had no significant effect on precontracted airway smooth muscle tone after muscarinic receptor blockade. These studies indicate that vagal preganglionic neurons are cholinergic in nature and coexpress VIP but do not contain NOS; their stimulation increases cholinergic outflow, without activation of inhibitory nonadrenergic, noncholinergic ganglionic neurons, stimulation of which induces airway smooth muscle relaxation. Furthermore, these studies do not support the possibility of direct inhibitory innervation of airway smooth muscle by vagal preganglionic fibers that contain VIP. PMID- 15358756 TI - Apoptosis in lung injury and remodeling. AB - The mode of cell death termed apoptosis, sometimes referred to as programmed cell death, is as critical a determinant of cell population size as is cell proliferation. Although best characterized in cells of the immune system, apoptosis is now known to be a key factor in the maintenance of normal cell turnover within structural cells in the parenchyma of virtually every organ. Recent interest in apoptosis in the lung has sparked a surge of investigations designed to determine the roles of apoptosis in lung development, injury, and remodeling. Of particular recent interest are the roles of apoptosis in disease pathogenesis and resolution, in which the concept of apoptosis as a "programmed" cell death, i.e., genetically determined, is often more accurately viewed as "inappropriate cell suicide" with regard to its extent and/or timing. Data accumulating over the past decade have made clear the complexity of the control of lung cell apoptosis; concepts of the regulation of apoptosis originally determined in classical cell culture models are often, but not always, applicable to structural cells. For this reason, each of the many cell types of the lung must be studied as a potentially new subject with its own idiosyncrasies yet to be discovered. In light of the large volume of literature now available, this article focuses on the roles of apoptosis in three pathophysiological contexts: acute respiratory distress syndrome, chronic obstructive pulmonary disease, and pulmonary fibrosis. Each section presents key data describing the evidence for apoptosis in the lung, its possible relevance to disease pathogenesis, and proposed mechanisms that might suggest potential avenues for therapeutic intervention. PMID- 15358757 TI - Alveolus formation: what have we learned from genetic studies? AB - The respiratory system has two basic functions: air exchange and pathogen clearance. The conducting airway and alveolar parenchyma are the basic structures to fulfill these functions during respiratory cycles. In humans, there are approximately 40 cell types in the lung that coordinately work together through various structural and signaling molecules. These molecules are vital for maintaining normal lung functions in response to environmental changes. Aberrant expression of these molecules can jeopardize human health and cause various pulmonary diseases. In this article, we will review some recent progress made in the pulmonary field, using genetic animal model systems to elucidate molecular mechanisms that are important for alveolar formation and lung diseases. PMID- 15358759 TI - Testosterone is responsible for enhanced susceptibility of males to ischemic renal injury. AB - Female mice are much more resistant to ischemia/reperfusion (I/R)-induced kidney injury when compared with males. Although estrogen administration can partially reduce kidney injury associated with I/R, we demonstrated that the presence of testosterone, more than the absence of estrogen, plays a critical role in gender differences in susceptibility of the kidney to ischemic injury. Testosterone administration to females increases kidney susceptibility to ischemia. Dihydrotestosterone, which can not be aromatized to estrogen, has effects equal to those of testosterone. Castration reduces the I/R-induced kidney injury. In contrast, ovariectomy does not affect kidney injury induced by ischemia in females. Testosterone reduces ischemia-induced activation of nitric oxide synthases (NOSs) and Akt and the ratio of extracellular signal related kinase (ERK) to c-jun N-terminal kinase (JNK) phosphorylation. Pharmacological (Nomega nitro-L-arginine) or genetic (endothelial NOS or inducible NOS) inhibition of NOSs in females enhances kidney susceptibility to ischemia. Nitric oxide increases Akt phosphorylation and protects Madin-Darby canine kidney epithelial cells from oxidant stress. Antagonists of androgen or estrogen receptors do not affect the gender differences. In conclusion, testosterone inhibits the post ischemic activation of NOSs and Akt and the ratio of ERK to JNK phosphorylation through non-androgen receptor-medicated mechanisms, leading to increased inflammation and increased functional injury to the kidney. These findings provide a new paradigm for the design of therapies for ischemia/reperfusion injury and may be important to our understanding of the pathophysiology of acute renal failure in pregnancy where plasma androgen levels are elevated. PMID- 15358760 TI - Sterol-responsive element-binding protein (SREBP) 2 down-regulates ATP-binding cassette transporter A1 in vascular endothelial cells: a novel role of SREBP in regulating cholesterol metabolism. AB - ATP-binding cassette transporter A1 (ABCA1) is a pivotal regulator of cholesterol efflux from cells to apolipoproteins, whereas sterol-responsive element-binding protein 2 (SREBP2) is the key protein regulating cholesterol synthesis and uptake. We investigated the regulation of ABCA1 by SREBP2 in vascular endothelial cells (ECs). Our results showed that sterol depletion activated SREBP2 and increased its target, low density lipoprotein receptor mRNA, with a concurrent decrease in the ABCA1 mRNA. Transient transfection analysis revealed that sterol depletion decreased the ABCA1 promoter activity by 50%, but low density lipoprotein receptor promoter- and the sterol-responsive element-driven luciferase activities were increased. Overexpression of the N terminus of SREBP2 (SREBP2(N)), an active form of SREBP2, also inhibited the ABCA1 promoter activity. Functionally adenovirus-mediated SREBP2(N) expression increased cholesterol accumulation and decreased apoA-I-mediated cholesterol efflux. The conserved E-box motif was responsible for the SREBP2(N)-mediated inhibition since mutation of the E-box increased the basal activity of the ABCA1 promoter and abolished the inhibitory effect of SREBP2(N). Furthermore sterol depletion and SREBP2(N) overexpression induced the binding of SREBP2(N) to both consensus and ABCA1-specific E-box. Chromatin immunoprecipitation assay demonstrated that serum starvation enhanced the association of SREBP2 and the ABCA1 promoter in ECs. To correlate this mechanism pathophysiologically, we found that oscillatory flow caused the activation of SREBP2 and therefore attenuated ABCA1 promoter activity in ECs. Thus, this SREBP-regulated mechanism may control the efflux of cholesterol, which is a newly defined function of SREBP2 in ECs in addition to its role in cholesterol uptake and biosynthesis. PMID- 15358761 TI - Translation of a yeast mitochondrial tRNA synthetase initiated at redundant non AUG codons. AB - Although initiation of translation at non-AUG codons occurs occasionally in prokaryotes and higher eukaryotes, it has not been reported in yeast until very recently. Evidence presented here shows that redundant ACG codons are recognized as alternative translation start sites for ALA1, the only gene in Saccharomyces cerevisiae coding for alanyl-tRNA synthetase. ALA1 is shown to be a bifunctional gene that provides both cytoplasmic and mitochondrial activities. Unlike most bifunctional genes that contain alternative in-frame AUG initiators, there is only one AUG codon, designated AUG1, close to the 5'-end of the ALA1 open reading frame. Transcriptional mapping identified three overlapping transcripts, with 5' ends at positions 54, 105, and 117 nucleotides upstream of AUG1, respectively. Site-specific mutagenesis demonstrated that the cytoplasmic and mitochondrial functions of ALA1 are provided by two protein isoforms with distinct amino termini; that is, a short cytoplasmic form initiated at AUG1 and a longer mitochondrial isoform initiated at two upstream in-frame ACG codons, i.e. ACG( 25) and ACG(-24). These two ACG codons function redundantly in initiation of translation. Either codon can function in the absence of the other. The short transcript appears to serve as the template for the cytoplasmic form, whereas the longer transcripts are likely to code for both isoforms via alternative initiation. Because yeast ribosomes in general cannot efficiently recognize a non AUG initiator, this unique feature of redundancy of non-AUG initiators in a single mRNA may in itself represent a novel paradigm for translation initiation from poor initiators. PMID- 15358762 TI - Substrate tRNA recognition mechanism of tRNA (m7G46) methyltransferase from Aquifex aeolicus. AB - Transfer RNA (m7G46) methyltransferase catalyzes the methyl transfer from S adenosylmethionine to N7 atom of the guanine 46 residue in tRNA. Analysis of the Aquifex aeolicus genome revealed one candidate open reading frame, aq065, encoding this gene. The aq065 protein was expressed in Escherichia coli and purified to homogeneity on 15% SDS-polyacrylamide gel electrophoresis. Although the overall amino acid sequence of the aq065 protein differs considerably from that of E. coli YggH, the purified aq065 protein possessed a tRNA (m7G46) methyltransferase activity. The modified nucleoside and its location were determined by liquid chromatography-mass spectroscopy. To clarify the RNA recognition mechanism of the enzyme, we investigated the methyl transfer activity to 28 variants of yeast tRNAPhe and E. coli tRNAThr. It was confirmed that 5' leader and 3'-trailer RNAs of tRNA precursor are not required for the methyl transfer. We found that the enzyme specificity was critically dependent on the size of the variable loop. Experiments using truncated variants showed that the variable loop sequence inserted between two stems is recognized as a substrate, and the most important recognition site is contained within the T stem. These results indicate that the L-shaped tRNA structure is not required for methyl acceptance activity. It was also found that nucleotide substitutions around G46 in three-dimensional core decrease the activity. PMID- 15358763 TI - Two homologous apolipoprotein AI mimetic peptides. Relationship between membrane interactions and biological activity. AB - Two related 18-amino acid, class A, amphipathic helical peptides termed 3F-2 and 3F14 were chosen for this study. Although they have identical amino acid compositions and many similar biophysical properties, 3F-2 is more potent than 3F14 as an apolipoprotein AI mimetic peptide. The two peptides exhibit similar gross conformational properties, forming structures of high helical content on a membrane surface. However, the thermal denaturation transition of 3F-2 is more cooperative, suggesting a higher degree of oligomerization on the membrane. Both 3F-2 and 3F14 promote the segregation of cholesterol in membranes containing phosphatidylcholine and cholesterol, but 3F-2 exhibits a greater selectivity for partitioning into cholesterol-depleted regions of the membrane. Magic angle spinning/NMR studies indicate that the aromatic residues of 3F-2 are stacked in the presence of lipid. The aromatic side chains of this peptide also penetrate more deeply into membranes of phosphatidylcholine with cholesterol compared with 3F14. Using the fluorescent probe, 1,3-dipyrenylpropane, we monitored the properties of the lipid hydrocarbon environment. 3F-2 had a greater effect in altering the properties of the hydrocarbon region of the membrane. The results are consistent with our proposed model of the effect of peptide shape on the nature of the difference in peptide insertion into the bilayer. PMID- 15358764 TI - Adenomatous polyposis coli control of retinoic acid biosynthesis is critical for zebrafish intestinal development and differentiation. AB - Mutations in the APC (adenomatous polyposis coli) tumor suppressor gene cause uncontrolled proliferation and impaired differentiation of intestinal epithelial cells. Recent studies indicate that human colon adenomas and carcinomas lack retinol dehydrogenases (RDHs) and that APC regulates the expression of human RDHL. These data suggest a model wherein APC controls enterocyte differentiation by controlling retinoic acid production. However, the importance of APC and retinoic acid in mediating control of normal enterocyte development and differentiation remains unclear. To examine the relationship between APC and retinoic acid biosynthesis in normal enterocytes, we have identified two novel zebrafish retinol dehydrogenases, termed zRDHA and zRDHB, that show strong expression within the gut of developing zebrafish embryos. Morpholino knockdown of either APC or zRDHB in zebrafish embryos resulted in defects in structures known to require retinoic acid. These defects included cardiac abnormalities, pericardial edema, failed jaw and pectoral fin development, and the absence of differentiated endocrine and exocrine pancreas. In addition, APC or zRDHB morphant fish developed intestines that lacked columnar epithelial cells and failed to express the differentiation marker intestinal fatty acid-binding protein. Treatment of either APC or zRDHB morphant embryos with retinoic acid rescued the defective phenotypes. Downstream of retinoic acid production, we identified hoxc8 as a retinoic acid-induced gene that, when ectopically expressed, rescued phenotypes of APC- and zRDHB-deficient zebrafish. Our data establish a genetic link supporting a critical role for retinoic acid downstream of APC and confirm the importance of retinoic acid in enterocyte differentiation. PMID- 15358765 TI - Invertebrate data predict an early emergence of vertebrate fibrillar collagen clades and an anti-incest model. AB - Fibrillar collagens are involved in the formation of striated fibrils and are present from the first multicellular animals, sponges, to humans. Recently, a new evolutionary model for fibrillar collagens has been suggested (Boot-Handford, R. P., Tuckwell, D. S., Plumb, D. A., Farrington Rock, C., and Poulsom, R. (2003) J. Biol. Chem. 278, 31067-31077). In this model, a rare genomic event leads to the formation of the founder vertebrate fibrillar collagen gene prior to the early vertebrate genome duplications and the radiation of the vertebrate fibrillar collagen clades (A, B, and C). Here, we present the modular structure of the fibrillar collagen chains present in different invertebrates from the protostome Anopheles gambiae to the chordate Ciona intestinalis. From their modular structure and the use of a triple helix instead of C-propeptide sequences in phylogenetic analyses, we were able to show that the divergence of A and B clades arose early during evolution because alpha chains related to these clades are present in protostomes. Moreover, the event leading to the divergence of B and C clades from a founder gene arose before the appearance of vertebrates; altogether these data contradict the Boot-Handford model. Moreover, they indicate that all the key steps required for the formation of fibrils of variable structure and functionality arose step by step during invertebrate evolution. PMID- 15358766 TI - The constitutive androstane receptor and pregnane X receptor function coordinately to prevent bile acid-induced hepatotoxicity. AB - A double null mouse line (2XENKO) lacking the xenobiotic receptors CAR (constitutive androstane receptor) (NR1I3) and PXR (pregnane X receptor) (NR1I2) was generated to study their functions in response to potentially toxic xenobiotic and endobiotic stimuli. Like the single knockouts, the 2XENKO mice are viable and fertile and show no overt phenotypes under normal conditions. As expected, they are completely insensitive to broad range xenobiotic inducers able to activate both receptors, such as clotrimazole and dieldrin. Comparisons of the single and double knockouts reveal specific roles for the two receptors. Thus, PXR does not contribute to the process of acetaminophen hepatotoxicity mediated by CAR, but both receptors contribute to the protective response to the hydrophobic bile acid lithocholic acid (LCA). As previously observed with PXR (Xie, W., Radominska-Pandya, A., Shi, Y., Simon, C. M., Nelson, M. C., Ong, E. S., Waxman, D. J., and Evans, R. M. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 3375-3380), pharmacologic activation of CAR induces multiple LCA detoxifying enzymes and provides strong protection against LCA toxicity. Comparison of their responses to LCA treatment demonstrates that CAR predominantly mediates induction of the cytochrome p450 CYP3A11 and the multidrug resistance-associated protein 3 transporter, whereas PXR is the major regulator of the Na+-dependent organic anion transporter 2. These differential responses may account for the significant sensitivity of the CAR knockouts, but not the PXR knockouts, to an acute LCA dose. Because this sensitivity is not further increased in the 2XENKO mice, CAR may play a primary role in acute responses to this toxic endobiotic. These results define a central role for CAR in LCA detoxification and show that CAR and PXR function coordinately to regulate both xenobiotic and bile acid metabolism. PMID- 15358767 TI - A DNA aptamer prevents influenza infection by blocking the receptor binding region of the viral hemagglutinin. AB - Influenza A virus infection is a major source of morbidity and mortality worldwide. Current means of control for influenza are based on prophylaxis by vaccines and on treatment by the available specific influenza neuraminidase inhibitor drugs. The approach taken in the present study is to prevent and/or ameliorate influenza infection by site-specific blocking of the viral binding to host cell receptors. We describe a novel oligonucleotide, known also as an aptamer, which has been designed to complement the receptor-binding region of the influenza hemagglutinin molecule. It was constructed by screening a DNA library and processing by the selective evolution of ligands by exponential enrichment (SELEX) procedure. We show that this DNA aptamer is indeed capable of inhibiting the hemagglutinin capacity of the virus, as well as in the prevention of viral infectivity in vitro, in tissue culture. Furthermore, it inhibits viral infection by different influenza strains in an animal model, as manifested by 90-99% reduction of virus burden in the lungs of treated mice. The mode of action of this aptamer is by blocking the binding of influenza virus to target cell receptors and consequently prevention of the virus invasion into the host cells. PMID- 15358768 TI - Autophosphorylation activity of the Arabidopsis ethylene receptor multigene family. AB - Receptors for the gaseous phytohormone ethylene show sequence similarity to bacterial two-component histidine kinases. These receptors are encoded by a multigene family that can be divided into subfamilies 1 and 2. It has been previously shown that a subfamily 1 Arabidopsis thaliana ethylene receptor, ETR1, autophosphorylates in vitro on a conserved histidine residue (1). However, sequence comparisons between the five ethylene receptor family members suggest that subfamily 2 members do not have all the motifs necessary for histidine kinase activity. Further, a tobacco subfamily 2 receptor, NTHK1, autophosphorylates on serines and threonines in vitro (2). Here we show that all five Arabidopsis ethylene receptor proteins autophosphorylate in vitro. We analyzed the nature of the phosphorylated amino acids by acid/base stability and bi-dimensional thin layer electrophoresis and demonstrated that unlike ETR1 all other ethylene receptors autophosphorylate predominantly on serine residues. ERS1, the only other subfamily 1 receptor, is able to phosphorylate on both histidine and serine residues in the presence of Mn2+. However, histidine autophosphorylation is lost when ERS1 is assayed in the presence of both Mg2+ and Mn2+, suggesting that this activity may not occur in vivo. Furthermore, mutation of the histidine residue conserved in two-component systems does not abolish serine autophosphorylation, eliminating the possibility of a histidine to serine phosphotransfer. Our biochemical observations complement the recently published genetic data that histidine kinase activity is not necessary for ethylene receptor function in plants and suggest that ethylene signal transduction does not occur through a phosphorelay mechanism. PMID- 15358769 TI - Hsp90 chaperones wild-type p53 tumor suppressor protein. AB - Immortalized human fibroblasts were used to investigate the putative interactions of the Hsp90 molecular chaperone with the wild-type p53 tumor suppressor protein. We show that geldanamycin or radicicol, specific inhibitors of Hsp90, diminish specific wild-type p53 binding to the p21 promoter sequence. Consequently, these inhibitors decrease p21 mRNA levels, which lead to a reduction in cellular p21/Waf1 protein, known to induce cell cycle arrest. In control experiments, we show that neither geldanamycin nor radicicol affect p53 mRNA levels. A minor decrease in p53 protein level following the treatment of human fibroblasts with the inhibitors suggests the potential involvement of Hsp90 in the stabilization of wild-type p53. To support our in vivo findings, we used a reconstituted system with highly purified recombinant proteins to examine the effects of Hsp90 on wild type p53 binding to the p21 promoter sequence. The human recombinant Hsp90 alpha isoform as well as bovine brain Hsp90 were purified to homogeneity. Both of these molecular chaperones displayed ATPase activity and the ability to refold heat inactivated luciferase in a geldanamycin- and radicicol-sensitive manner, suggesting that post-translational modifications are not involved in the modulation of Hsp90alpha activity. We show that the incubation of recombinant p53 at 37 degrees C decreases the level of its wild-type conformation and strongly inhibits the in vitro binding of p53 to the p21 promoter sequence. Interestingly, Hsp90 in an ATP-dependent manner can positively modulate p53 DNA binding after incubation at physiological temperature of 37 degrees C. Other recombinant human chaperones from Hsp70 and Hsp40 families were not able to efficiently substitute Hsp90 in this reaction. Consistent with our in vivo results, geldanamycin can suppress Hsp90 ability to regulate in vitro p53 DNA binding to the promoter sequence. In summary, the results presented in this article state that chaperone activity of Hsp90 is important for the transcriptional activity of genotypically wild-type p53. PMID- 15358770 TI - Desulfoglucosinolate sulfotransferases from Arabidopsis thaliana catalyze the final step in the biosynthesis of the glucosinolate core structure. AB - The phytotoxin coronatine is a structural analog of octadecanoid signaling molecules, which are well known mediators of plant defense reactions. To isolate novel coronatine-regulated genes from Arabidopsis thaliana, differential mRNA display was performed. Transcript levels of CORI-7 (coronatine induced-7) were rapidly and transiently increased in coronatine-treated plants, and the corresponding cDNA was found to encode the sulfotransferase AtST5a. Likewise, upon wounding, an immediate and transient increase in AtST5a mRNA levels could be observed in both locally wounded and unwounded (systemic) leaves. Furthermore, application of octadecanoids and ethylene as compounds involved in plant wound defense reactions resulted in AtST5a gene activation, whereas pathogen defense related signals (yeast elicitor and salicylic acid) were inactive. AtST5a and its close homologs AtST5b and AtST5c were purified as His6-tagged proteins from Escherichia coli. The three enzymes were shown to catalyze the final step in the biosynthesis of the glucosinolate (GS) core structure, the sulfation of desulfoglucosinolates (dsGSs). They accept a broad range of dsGSs as substrates. However, in a competitive situation, AtST5a clearly prefers tryptophan- and phenylalanine-derived dsGSs, whereas long chain dsGSs derived from methionine are the preferred substrates of AtST5b and AtST5c. Treatment of Arabidopsis plants with low concentrations of coronatine resulted in an increase in the amounts of specific GSs, primarily glucobrassicin and neoglucobrassicin. Hence, it is suggested that AtST5a is the sulfotransferase responsible for the biosynthesis of tryptophan-derived GSs in vivo. PMID- 15358771 TI - Hsp90 regulates the activity of wild type p53 under physiological and elevated temperatures. AB - The activity and structural integrity of the tumor suppressor protein p53 is of crucial importance for the prevention of cancer. p53 is a conformational flexible and labile protein, in which structured and unstructured regions function in a synergistic manner. The molecular chaperone Hsp90 is known to bind to mutant and wild type p53 in vivo. Using highly purified proteins we analyzed the interaction and the binding sites between both proteins in detail. Our results demonstrate that Hsp90 binds to a folded, native-like conformation of p53 in vitro with micromolar affinity. Specifically, the DNA-binding domain of p53 and the middle and carboxy-terminal domains of Hsp90 are responsible for this interaction, which is essential to stabilize p53 at physiological temperatures and to prevent it from irreversible thermal inactivation. Our results are in agreement with a model in which Hsp90 is required to maintain the folded, active state of p53 by a reversible interaction, thus introducing an additional level of regulation. PMID- 15358772 TI - Cloning and characterization of the pheromone biosynthesis activating neuropeptide receptor from the silkmoth, Bombyx mori. Significance of the carboxyl terminus in receptor internalization. AB - In most Lepidoptera, pheromone biosynthesis is regulated by a neuropeptide termed pheromone biosynthesis activating neuropeptide (PBAN). Although much is known about the cellular targets of PBAN, identification and functional characterization of the PBAN receptor (PBANR) has proven to be elusive. Given the sequence similarity between the active C-terminal regions of PBAN and neuromedin U, it was hypothesized that their respective receptors might also be similar in structure (Park, Y., Kim, Y. J., and Adams, M. E. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 11423-11428). Consequently, utilizing primers constructed from the conserved regions of insect neuromedin U receptor homologues, a full-length 2780 nucleotide clone encoding a 46-kDa G protein-coupled receptor was amplified from a Bombyx mori pheromone gland cDNA library. Tissue distribution analyses revealed that the receptor transcript is specific to the pheromone gland where it undergoes significant up-regulation in the day preceding eclosion. When transiently expressed in Sf9 cells, the B. mori PBANR responds to PBAN by mobilizing extracellular calcium in a dose-dependent manner. Confocal microscopic studies demonstrated the specificity of enhanced green fluorescent protein-tagged B. mori PBANR for PBAN and showed that PBAN induces internalization of the PBANR.PBAN complex. The rapid onset of internalization is mediated by a 67-amino acid C-terminal extension absent in the cloned Helicoverpa zea PBANR, which suggests that receptor internalization in that species likely utilizes a different mechanism. From these results, we have concluded that the cloned receptor gene encodes the B. mori PBANR and that it is both structurally and functionally distinct from the H. zea PBANR. PMID- 15358773 TI - Autophagy gene disruption reveals a non-vacuolar cell death pathway in Dictyostelium. AB - Types of cell death include apoptosis, necrosis, and autophagic cell death. The latter can be defined as death of cells containing autophagosomes, autophagic bodies, and/or vacuoles. Are autophagy and vacuolization causes, consequences, or side effects in cell death with autophagy? Would control of autophagy suffice to control this type of cell death? We disrupted the atg1 autophagy gene in Dictyostelium discoideum, a genetically tractable model for developmental autophagic vacuolar cell death. The procedure that induced autophagy, vacuolization, and death in wild-type cells led in atg1 mutant cells to impaired autophagy and to no vacuolization, demonstrating that atg1 is required for vacuolization. Unexpectedly, however, cell death still took place, with a non vacuolar and centrally condensed morphology. Thus, a cell death mechanism that does not require vacuolization can operate in this cell death model showing conspicuous vacuolization. The revelation of non-vacuolar cell death in this protist by autophagy gene disruption is reminiscent of caspase inhibition revealing necrotic cell death in animal cells. Thus, hidden alternative cell death pathways may be found across kingdoms and for diverse types of cell death. PMID- 15358774 TI - The co-activator CREB-binding protein participates in enhancer-dependent activities of bicoid. AB - Bicoid (Bcd) is a transcriptional activator required for early embryonic patterning in Drosophila. Despite extensive studies, it currently remains unclear how Bcd activates transcription and what proteins participate in its activation process. In this report, we describe experiments to analyze the role of the Drosophila co-activator dCBP in Bcd-mediated activation. In Drosophila S2 cells, the Bcd activity is increased by the co-transfection of plasmids expressing dCBP and reduced by double-stranded RNA-mediated interference against dCBP. We further show that Bcd and dCBP can interact with each other and that Bcd-interacting domains of dCBP can cause dominant negative effects on Bcd activity in S2 cells. Our comparison of two Bcd-responsive enhancers, hunchback (hb) and knirps (kni), reveals a differential role of dCBP in facilitating Bcd activation. A dCBP mutant defective in its histone acetyltransferase activity exhibits a reduced, but not abolished, co-activator function for Bcd. Our chromatin immunoprecipitation experiments show that dCBP can increase not only the occupancy of Bcd itself at the enhancers but also the recruitment of general transcription factors to the promoter. Together, these experiments suggest that dCBP is an enhancer-dependent co-activator of Bcd, facilitating its activation through multiple mechanisms. PMID- 15358775 TI - Interaction with cystic fibrosis transmembrane conductance regulator-associated ligand (CAL) inhibits beta1-adrenergic receptor surface expression. AB - G protein-coupled receptors such as the beta1-adrenergic receptor (beta1AR) must be trafficked to the plasma membrane in order to bind with their extracellular ligands and regulate cellular physiology. By using glutathione S-transferase pull down techniques, we found that the beta1AR carboxyl terminus directly interacts with the cystic fibrosis transmembrane conductance regulator-associated ligand (CAL; also known as PIST, GOPC, and FIG), a protein known to be primarily localized to the Golgi apparatus. CAL contains two predicted coiled-coil domains and one PSD-95/Discs-large/ZO-1 homology (PDZ) domain. The beta1AR carboxyl terminus (CT) binds to the PDZ domain of CAL, with the last few amino acids (ESKV) of the beta1AR-CT being the key determinants for the interaction. Mutation of the terminal valine residue resulted in markedly reduced association of the beta1AR-CT with CAL. Numerous other mutations to the ESKV motif also impaired the beta1AR-CT/CAL interaction, suggesting that this motif is close to optimal for association with the CAL PDZ domain. In cells, full-length beta1AR robustly associates with CAL, and this interaction is abolished by mutation of the terminal valine to alanine of the receptor (V477A), as determined by co immunoprecipitation experiments and immunofluorescence co-localization studies. Consistent with observations that CAL is a Golgi-associated protein, overexpression of CAL reduces surface expression of beta1AR. Interaction with CAL promotes retention of beta1AR within the cell, whereas PSD-95, another beta1AR associated PDZ domain-containing protein, competitively blocks beta1AR association with CAL and promotes receptor trafficking to the cell surface. These data reveal that CAL, a novel beta1AR-binding partner, modulates beta1AR intracellular trafficking, thereby revealing a new mechanism of regulation for beta1AR anterograde trafficking through the endoplasmic reticulum-Golgi complex to the plasma membrane. PMID- 15358776 TI - Cooperative conformational changes in a G-protein-coupled receptor dimer, the leukotriene B(4) receptor BLT1. AB - We have used an isolated receptor, the leukotriene B(4) receptor BLT1, to analyze the mechanism of receptor activation in a G-protein-coupled receptor dimer. The isolated receptor is essentially a dimer whether the agonist is present or not, provided the detergent used stabilizes the inactive dimeric assembly. We have produced a receptor mutant where Cys(97) in the third transmembrane domain has been replaced by a serine. This mutation leads to an approximately 100-fold decrease in the affinity for the agonist. 5-Hydroxytryptophan has then been introduced at position 234 in the C97A mutant sixth transmembrane domain. Agonist binding to the labeled receptor is associated with variations in the fluorescence properties of 5-hydroxytryptophan due to specific agonist-induced conformational changes. The C97A mutant labeled with 5-hydroxytryptophan has then been associated with a wild-type receptor in a dimeric complex that has been subsequently purified. The purified complex activates its G-protein partner in a similar manner as the wild-type homodimer. Due to the difference in the affinity for the agonist between the wild-type and mutant protomers in this dimer, we have been able to reach a state where one of the protomers, the mutant, is in its unliganded state, whereas the other, the wild type, is loaded with the agonist. We show that agonist binding to the wild-type receptor induces specific changes in the conformation of the unliganded protomer, as evidenced by the variations in the emission of the 5-hydroxytryptophan residue in the mutant receptor. These data provide a direct demonstration for agonist-induced cooperative conformational changes in a GPCR dimer. PMID- 15358777 TI - Diphenyleneiodonium inhibits the cell redox metabolism and induces oxidative stress. AB - Diphenyleneiodonium (DPI) and the structurally related compound diphenyliodonium (DIP) are widely used as inhibitors of flavoenzymes, particularly NADPH oxidase. Here we report further evidence that DPI and DIP are not specific flavin binders. A 3-h incubation of N11 glial cells with DPI significantly inhibited in a dose dependent way both the pentose phosphate pathway and the tricarboxylic acid cycle. In parallel, we observed a dose-dependent increase of reactive oxygen species generation and lipoperoxidation and increased leakage of lactate dehydrogenase activity in the extracellular medium. The glutathione/glutathione disulfide ratio decreased, whereas the efflux of glutathione out of the cells increased. This suggests that DPI causes an augmented oxidative stress and exerts a cytotoxic effect in N11 cells. Indeed, the cells were protected from these events when loaded with glutathione. Similar results were observed using DIP instead of DPI and also in other cell types. We suggest that the DPI-elicited inhibition of the pentose phosphate pathway and tricarboxylic acid cycle may be mediated by the blockade of several NAD(P)-dependent enzymes, such as glucose 6 phosphate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase, and lactate dehydrogenase. In light of these results, we think that some effects of DPI or DIP in in vitro and in vivo experimental models should be interpreted with caution. PMID- 15358778 TI - The acidic C-terminal domain stabilizes the chaperone function of protein disulfide isomerase. AB - Protein disulfide isomerase (PDI, EC 5.3.4.1) is a chaperone and catalyzes the formation and rearrangement of disulfide bonds in proteins. Domain c-(463-491), containing 18 acidic residues, is an interesting and important C-terminal extension of PDI. In this study, the PDI mutant abb'a', in which domain c is truncated, was used to investigate the relationship between the C-terminal structure and chaperone function. Reactivation and light-scattering experiments show that both wild-type PDI and abb'a' interact with lactate dehydrogenase (LDH, EC 1.1.1.27), which tends to self-aggregate during reactivation. The interaction enhances reactivation of LDH and reduces aggregation. According to these results, it seems as if domain c might be dispensable to the chaperone function of PDI. However, abb'a' is prone to self-aggregation and causes increased aggregation of LDH during thermal denaturation. In contrast, wild-type PDI remains active as a chaperone under these conditions and prevents self-aggregation of LDH. Furthermore, measurements of intrinsic fluorescence and difference absorbance during denaturation show that abb'a' is much more labile to heat or guanidine hydrochloride denaturation than wild-type PDI. This suggests that domain c is required for the stabilization and maintenance of the chaperone function of PDI under extreme conditions. PMID- 15358779 TI - Cardiac troponin T isoforms affect the Ca(2+) sensitivity of force development in the presence of slow skeletal troponin I: insights into the role of troponin T isoforms in the fetal heart. AB - In this study we investigated the physiological role of the cardiac troponin T (cTnT) isoforms in the presence of human slow skeletal troponin I (ssTnI). ssTnI is the main troponin I isoform in the fetal human heart. In reconstituted fibers containing the cTnT isoforms in the presence of ssTnI, cTnT1-containing fibers showed increased Ca(2+) sensitivity of force development compared with cTnT3- and cTnT4-containing fibers. The maximal force in reconstituted skinned fibers was significantly greater for the cTnT1 (predominant fetal cTnT isoform) when compared with cTnT3 (adult TnT isoform) in the presence of ssTnI. Troponin (Tn) complexes containing ssTnI and reconstituted with cTnT isoforms all yielded different maximal actomyosin ATPase activities. Tn complexes containing cTnT1 and cTnT4 (both fetal isoforms) had a reduced ability to inhibit actomyosin ATPase activity when compared with cTnT3 (adult isoform) in the presence of ssTnI. The rate at which Ca(2+) was released from site II of cTnC in the cTnI.cTnC complex (122/s) was 12.5-fold faster than for the ssTnI.cTnC complex (9.8/s). Addition of cTnT3 to the cTnI.cTnC complex resulted in a 3.6-fold decrease in the Ca(2+) dissociation rate from site II of cTnC. Addition of cTnT3 to the ssTnI.cTnC complex resulted in a 1.9-fold increase in the Ca(2+) dissociation rate from site II of cTnC. The rate at which Ca(2+) dissociated from site II of cTnC in Tn complexes also depended on the cTnT isoform present. However, the TnI isoforms had greater effects on the Ca(2+) dissociation rate of site II than the cTnT isoforms. These results suggest that the different N-terminal TnT isoforms would produce distinct functional properties in the presence of ssTnI when compared with cTnI and that each isoform would have a specific physiological role in cardiac muscle. PMID- 15358780 TI - Zinc potentiates an uncoupled anion conductance associated with the dopamine transporter. AB - Binding of Zn(2+) to an endogenous binding site in the dopamine transporter (DAT) leads to inhibition of dopamine (DA) uptake and enhancement of carrier-mediated substrate efflux. To elucidate the molecular mechanism for this dual effect, we expressed the DAT and selected mutants in Xenopus laevis oocytes and applied the two-electrode voltage clamp technique together with substrate flux studies employing radiolabeled tracers. Under voltage clamp conditions we found that Zn(2+) (10 mum) enhanced the current induced by both DA and amphetamine. This was not accompanied by a change in the uptake rate but by a marked increase in the charge/DA flux coupling ratio as assessed from concomitant measurements of [(3)H]DA uptake and currents in voltage-clamped oocytes. These data suggest that Zn(2+) facilitates an uncoupled ion conductance mediated by DAT. Whereas this required substrate in the wild type (WT), we observed that Zn(2+) by itself activated such a conductance in a previously described mutant (Y335A). This signifies that the conductance is not strictly dependent on an active transport process. Ion substitution experiments in Y335A, as well as in WT, indicated that the uncoupled conductance activated by Zn(2+) was mainly carried by Cl(-). Experiments in oocytes under non-voltage-clamped conditions revealed furthermore that Zn(2+) could enhance the depolarizing effect of substrates in oocytes expressing WT. The data suggest that by potentiating an uncoupled Cl(-) conductance, Zn(2+) is capable of modulating the membrane potential of cells expressing DAT and as a result cause simultaneous inhibition of uptake and enhancement of efflux. PMID- 15358781 TI - MLE functions as a transcriptional regulator of the roX2 gene. AB - Dosage compensation is a process that equalizes transcription activity between the sexes. In Drosophila, two non-coding RNA, roX1 and roX2, and at least six protein regulators, MSL-1, MSL-2, MSL-3, MLE, MOF, and JIL-1, have been identified as essential for dosage compensation. Although there is accumulating evidence of the intricate functional and physical interactions between protein and RNA regulators, little is known about how roX RNA expression and function are modulated in coordination with protein regulators. In this report, we have found that a relatively short (about 350 bp) upstream genomic region of the roX2 gene, Prox2, harbors an activity that drives transcription of the downstream gene. Our study has shown that MLE can stimulate the transcription activity of Prox2 and that MLE associates with Prox2 through direct interaction with a newly identified 54-bp repeat, Prox. Our observations suggest a novel mechanism by which roX2 RNA is regulated at the transcriptional level. PMID- 15358782 TI - A dynamic pool of calcium in catecholamine storage vesicles. Exploration in living cells by a novel vesicle-targeted chromogranin A-aequorin chimeric photoprotein. AB - Chromaffin vesicles contain very high concentration of Ca2+ (approximately 20-40 mM total), compared with approximately 100 nM in the cytosol. Aequorin, a jellyfish photoprotein with Ca(2+)-dependent luminescence, measures [Ca2+] in specific subcellular compartments wherein proteins with organelle-specific trafficking domains are fused in-frame to aequorin. Because of the presence of vesicular trafficking domain within CgA we engineered sorting of an expressed human CgA-Aequorin fusion protein (hCgA-Aeq) into the vesicle compartment as confirmed by sucrose density gradients and confocal immunofluorescent co localization studies. hCgA-Aeq and cytoplasmic aequorin (Cyto-Aeq) luminescence displayed linear functions of [Ca2+] in vitro, over >5 log10 orders of magnitude (r > 0.99), and down to at least 10(-7) M sensitivity. Calibrating the pH dependence of hCgA-Aeq luminescence allowed estimation of [Ca2+]ves at granule interior pH (approximately 5.5). In the cytoplasm, Cyto-Aeq accurately determined [Ca2+]cyto under both basal ([Ca2+]cyto = 130 +/- 35 nM) and exocytosis stimulated conditions, confirmed by an independent reference technique (Indo-1 fluorescence). The hCgA-Aeq chimera determined vesicular free [Ca2+]ves = 1.4 +/- 0.3 microM under basal conditions indicating that >99% of granule total Ca2+ is in a "bound" state. The basal free [Ca2+]ves/[Ca2+]cyto ratio was thus approximately 10.8-fold, indicating active, dynamic Ca2+ uptake from cytosol into the granules. Stimulation of exocytotic secretion revealed prompt, dynamic increases in both [Ca2+](ves) and [Ca2+]cyto, and an exponential relation between the two (y = 0.99 x e(1.53x), r = 0.99), reflecting a persistent [Ca2+]ves/[Ca2+]cyto gradient, even during sharp increments of both values. Studies with inhibitors of Ca2+ translocation (Ca(2+)-ATPase), Na+/Ca(+) exchange, Na+/H(+)-exchange, and vesicle acidification (H(+)-translocating ATPase), documented a role for these four ion transporter classes in accumulation of Ca2+ inside the vesicles. PMID- 15358783 TI - Identification of all-trans-retinol:all-trans-13,14-dihydroretinol saturase. AB - Retinoids carry out essential functions in vertebrate development and vision. Many of the retinoid processing enzymes remain to be identified at the molecular level. To expand the knowledge of retinoid biochemistry in vertebrates, we studied the enzymes involved in plant metabolism of carotenoids, a related group of compounds. We identified a family of vertebrate enzymes that share significant similarity and a putative phytoene desaturase domain with a recently described plant carotenoid isomerase (CRTISO), which isomerizes prolycopene to all-trans lycopene. Comparison of heterologously expressed mouse and plant enzymes indicates that unlike plant CRTISO, the CRTISO-related mouse enzyme is inactive toward prolycopene. Instead, the CRTISO-related mouse enzyme is a retinol saturase carrying out the saturation of the 13-14 double bond of all-trans retinol to produce all-trans-13,14-dihydroretinol. The product of mouse retinol saturase (RetSat) has a shifted UV absorbance maximum, lambda(max) = 290 nm, compared with the parent compound, all-trans-retinol (lambda(max) = 325 nm), and its MS analysis (m/z = 288) indicates saturation of a double bond. The product was further identified as all-trans-13,14-dihydroretinol, since its characteristics were identical to those of a synthetic standard. Mouse RetSat is membrane-associated and expressed in many tissues, with the highest levels in liver, kidney, and intestine. All-trans-13,14-dihydroretinol was also detected in several tissues of animals maintained on a normal diet. Thus, saturation of all trans-retinol to all-trans-13,14-dihydroretinol by RetSat produces a new metabolite of yet unknown biological function. PMID- 15358784 TI - Conservation of Bmp2 post-transcriptional regulatory mechanisms. AB - Bone morphogenetic protein (BMP) orthologs from diverse species like flies and humans are functionally interchangeable and play key roles in fundamental processes such as dorso-ventral axis formation in metazoans. Because both transcriptional and post-transcriptional mechanisms play central roles in modulating developmental protein levels, we have analyzed the 3'-untranslated region (3'UTR) of the Bmp 2 gene. This 3'UTR is unusually long and is alternatively polyadenylated. Mouse, human, and dog mRNAs are 83-87% identical within this region. A 265-nucleotide sequence, conserved between mammals, birds, frogs, and fish, is present in Bmp2 but not Bmp4. The ability of AmphiBMP2/4, a chordate ortholog to Bmp2 and Bmp4, to align with this sequence suggests that its function may have been lost in Bmp4. Activation of reporter genes by the conserved region acts by a post-transcriptional mechanism. Mouse, human, chick, and zebrafish Bmp2 synthetic RNAs decay rapidly in extracts from cells not expressing Bmp2. In contrast, these RNAs are relatively stable in extracts from Bmp2-expressing cells. Thus, Bmp2 RNA half-lives in vitro correlate with natural Bmp2 mRNA levels. The fact that non-murine RNAs interact appropriately with the mouse decay machinery suggests that the function of these cis-regulatory regions has been conserved for 450 million years since the fish and tetrapod lineages diverged. Overall, our results suggest that the Bmp2 3'UTR contains essential regulatory elements that act post-transcriptionally. PMID- 15358785 TI - NARC-1/PCSK9 and its natural mutants: zymogen cleavage and effects on the low density lipoprotein (LDL) receptor and LDL cholesterol. AB - The discovery of autosomal dominant hypercholesterolemic patients with mutations in the PCSK9 gene, encoding the proprotein convertase NARC-1, resulting in the missense mutations suggested a role in low density lipoprotein (LDL) metabolism. We show that the endoplasmic reticulum-localized proNARC-1 to NARC-1 zymogen conversion is Ca2+-independent and that within the zymogen autocatalytic processing site SSVFAQ [downward arrow]SIP Val at P4 and Pro at P3' are critical. The S127R and D374Y mutations result in approximately 50-60% and > or =98% decrease in zymogen processing, respectively. In contrast, the double [D374Y + N157K], F216L, and R218S natural mutants resulted in normal zymogen processing. The cell surface LDL receptor (LDLR) levels are reduced by 35% in lymphoblasts of S127R patients. The LDLR levels are also reduced in stable HepG2 cells overexpressing NARC-1 or its natural mutant S127R, and this reduction is abrogated in the presence of 5 mm ammonium chloride, suggesting that overexpression of NARC-1 increases the turnover rate of the LDLR. Adenoviral expression of wild type human NARC-1 in mice resulted in a maximal approximately 9-fold increase in circulating LDL cholesterol, while in LDLR-/- mice a delayed approximately 2-fold increase in LDL cholesterol was observed. In conclusion, NARC-1 seems to affect both the level of LDLR and that of circulating apoB containing lipoproteins in an LDLR-dependent and -independent fashion. PMID- 15358786 TI - UV-induced expression of key component of the tanning process, the POMC and MC1R genes, is dependent on the p-38-activated upstream stimulating factor-1 (USF-1). AB - Protection against UV-mediated DNA damage and the onset of oncogenesis is afforded by the tanning response in which UV irradiation triggers melanocytes to increase production of melanin that is then transferred to keratinocytes. A key component of the tanning process is the UV-mediated induction of the pro opiomelanocortin (POMC) and MC1R genes encoding the alpha-melanocyte-stimulating hormone and its receptor, respectively, which play a crucial role in pigmentation by regulating the intracellular levels of cAMP. How these genes are regulated in response to UV irradiation is not known. Here we have shown that UV-induced activation of the POMC and MC1R promoters is mediated by p38 stress-activated kinase signaling to the transcription factor, upstream stimulating factor-1 (USF 1). Importantly, melanocytes derived from USF-1 -/- mice exhibit a defective UV response and fail to activate POMC and MC1R expression in response to UV irradiation. The results define USF-1 as a critical UV-responsive activator of genes implicated in protection from solar radiation. PMID- 15358787 TI - The adaptive response to dietary zinc in mice involves the differential cellular localization and zinc regulation of the zinc transporters ZIP4 and ZIP5. AB - The ZIP5 gene encodes a protein closely related to ZIP4, a zinc transporter mutated in the human genetic disorder acrodermatitis enteropathica. Herein, we demonstrate that mouse ZIP5 and ZIP4 genes are co-expressed in several tissues involved in zinc homeostasis (intestine, pancreas, embryonic yolk sac). However, unlike expression of the ZIP4 gene, which is induced during periods of zinc deficiency, ZIP5 gene expression is unaltered by dietary zinc. Immunohistochemistry localizes ZIP5 to the basolateral surfaces of enterocytes, acinar cells, and visceral endoderm cells in mice fed a zinc-adequate diet. However, this protein is removed from these cell surfaces and internalized during dietary zinc deficiency. In contrast, ZIP4 is induced and recruited to the apical surface of enterocytes and endoderm cells during zinc deficiency. In the pancreas, ZIP4 is expressed in beta-cells, whereas ZIP5 is expressed in acinar cells. These results suggest that the function of ZIP5 is antagonistic to that of ZIP4 in the control of zinc homeostasis; rather than functioning in the acquisition of dietary zinc, as does ZIP4, ZIP5 may function in the removal of zinc from the body. Thus, during periods when dietary zinc is replete, ZIP5 may function to remove zinc from the blood via the pancreas and intestine, the major sites of zinc excretion in mammals, whereas the acquisition of dietary zinc by intestinal ZIP4 would be minimal. In contrast, during periods of dietary zinc deficiency when secretion of zinc by the pancreas and intestine is minimized, ZIP5 is removed from the cell surface, and the intestinal uptake of zinc is augmented by induction of ZIP4. PMID- 15358788 TI - Coordination of divalent metal ions in the active site of poly(A)-specific ribonuclease. AB - Poly(A)-specific ribonuclease (PARN) is a highly poly(A)-specific 3' exoribonuclease that efficiently degrades mRNA poly(A) tails. PARN belongs to the DEDD family of nucleases, and four conserved residues are essential for PARN activity, i.e. Asp-28, Glu-30, Asp-292, and Asp-382. Here we have investigated how catalytically important divalent metal ions are coordinated in the active site of PARN. Each of the conserved amino acid residues was substituted with cysteines, and it was found that all four mutants were inactive in the presence of Mg2+. However, in the presence of Mn2+, Zn2+, Co2+, or Cd2+, PARN activity was rescued from the PARN(D28C), PARN(D292C), and PARN(D382C) variants, suggesting that these three amino acids interact with catalytically essential metal ions. It was found that the shortest sufficient substrate for PARN activity was adenosine trinucleotide (A3) in the presence of Mg2+ or Cd2+. Interestingly, adenosine dinucleotide (A) was efficiently hydrolyzed in the presence of Mn2+, Zn2+, or Co2+, suggesting that the substrate length requirement for PARN can be modulated by the identity of the divalent metal ion. Finally, introduction of phosphorothioate modifications into the A substrate demonstrated that the scissile bond non-bridging phosphate oxygen in the pro-R position plays an important role during cleavage, most likely by coordinating a catalytically important divalent metal ion. Based on our data we discuss binding and coordination of divalent metal ions in the active site of PARN. PMID- 15358789 TI - Degradation of the gluconeogenic enzymes fructose-1,6-bisphosphatase and malate dehydrogenase is mediated by distinct proteolytic pathways and signaling events. AB - The key gluconeogenic enzyme fructose-1,6-bisphosphatase (FBPase) is subjected to catabolite inactivation and degradation when glucose-starved cells are replenished with fresh glucose. In various studies, the proteasome and the vacuole have each been reported to be the major site of FBPase degradation. Because different growth conditions were used in these studies, we examined whether variations in growth conditions could alter the site of FBPase degradation. Here, we demonstrated that FBPase was degraded outside the vacuole (most likely in the proteasome), when glucose was added to cells that were grown in low glucose media for a short period of time. By contrast, cells that were grown in the same low glucose media for longer periods of time degraded FBPase in the vacuole in response to glucose. Another gluconeogenic enzyme malate dehydrogenase (MDH2) showed the same degradation characteristics as FBPase in that the short term starvation of cells led to a non-vacuolar degradation, whereas long term starvation resulted in the vacuolar degradation of this protein. The N-terminal proline is required for the degradation of FBPase and MDH2 for both the vacuolar and non-vacuolar proteolytic pathways. The cAMP signaling pathway and the phosphorylation of glucose were needed for the vacuolar dependent degradation of FBPase and MDH2. By contrast, the cAMP-dependent signaling pathway was not involved in the non-vacuolar degradation of these proteins, although the phosphorylation of glucose was required. PMID- 15358790 TI - Biochemical and biological characterization of a novel anti-aromatase coumarin derivative. AB - Estrogen stimulates the proliferation of estrogen receptor (ER)-positive breast cancer cells. Aromatase is the enzyme responsible for the conversion of androgens into estrogens, and synthetic aromatase inhibitors such as letrozole, anastrozole, and exemestane have proven to be effective endocrine regimens for ER positive breast cancer. In a recent study, we have found that 4-benzyl-3-(4' chlorophenyl)-7-methoxycoumarin is a potent competitive inhibitor of aromatase with respect to the androgen substrate. Its K(i) value was determined to be 84 nm, significantly more potent than several known aromatase inhibitors. The specific interaction of this compound with aromatase was further demonstrated by the reduction of its binding by several mutations at the active site region of aromatase and evaluated by computer modeling analysis. The structure-activity studies have revealed that three functional groups (i.e. 3-(4'-chlorophenyl), 4 benzyl, and 7-methoxyl) of this coumarin are important in its inhibition of aromatase. In addition, through a matrigel thread three-dimensional cell culture, this compound was shown to behave like known aromatase inhibitors that suppress the proliferation of aromatase and estrogen receptor positive MCF-7aro breast cancer cells. This coumarin has been shown not to be cytotoxic at up to 40 mum. It was found not to be an inhibitor of steroid 5alpha-reductase that also utilizes androgen as the substrate and not to be a ligand of ERalpha, ERbeta, estrogen-related receptors, or androgen receptor. These results demonstrate that coumarins (a common type of phytochemical) or their derivatives can be potent inhibitors of aromatase and may be useful in suppressing aromataseand ER-positive breast tumors. PMID- 15358791 TI - Identifying structural features of fibrillar islet amyloid polypeptide using site directed spin labeling. AB - Pancreatic amyloid deposits, composed primarily of the 37-residue islet amyloid polypeptide (IAPP), are a characteristic feature found in more than 90% of patients with type II diabetes. Although IAPP amyloid deposits are associated with areas of pancreatic islet beta-cell dysfunction and depletion and are thought to play a role in disease, their structure is unknown. We used electron paramagnetic resonance spectroscopy to analyze eight spin-labeled derivatives of IAPP in an effort to determine structural features of the peptide. In solution, all eight derivatives gave rise to electron paramagnetic resonance spectra with sharp lines indicative of rapid motion on the sub-nanosecond time scale. These spectra are consistent with a rapidly tumbling and highly dynamic peptide. In contrast, spectra for the fibrillar form exhibit reduced mobility and the presence of strong intermolecular spin-spin interactions. The latter implies that the peptide subunits are ordered and that the same residues from neighboring peptides are in close proximity to one another. Our data are consistent with a parallel arrangement of IAPP peptides within the amyloid fibril. Analysis of spin label mobility indicates a high degree of order throughout the peptide, although the N-terminal region is slightly less ordered. Possible similarities with respect to the domain organization and parallelism of Alzheimer's amyloid beta peptide fibrils are discussed. PMID- 15358792 TI - Change in object naming ability during adulthood. AB - Using longitudinal data on the Boston Naming Test ( Kaplan, Goodglass, & Weintraub, 1983) collected over 20 years from healthy individuals aged 30 to 94, we examined change in lexical retrieval with age, gender, education, and their interactions. We compared results between random-effects longitudinal and traditional cross-sectional models. Random-effects modeling revealed significant linear and quadratic change in lexical retrieval with age; it also showed a Gender x Education interaction, indicating poorest performance for women with less education. Cross-sectional analyses produced greater estimates of change with age than did longitudinal analyses. PMID- 15358793 TI - Age differences in central (semantic) and peripheral processing: the importance of considering both response times and errors. AB - In this project we examined the effect of adult age on visual word recognition by using combined reaction time (RT) and accuracy methods based on the Hick-Hyman law. This was necessary because separate Brinley analyses of RT and errors resulted in contradicting results. We report the results of a lexical decision task experiment (with 96 younger adults and 97 older adults). We transformed the error data into entropy and then predicted RT by using entropy values separately for exposure duration (thought to influence peripheral processes) and word frequency (thought to influence central processes). For exposure duration, the entropy-RT functions indicate that older adults show higher intercepts and slopes than do younger adults, suggesting an encoding decrement for older adults. However, for word frequency, older adults show higher intercepts but not steeper slopes than younger adults. Older adults thus show a peripheral processing decrement but not a central processing decrement for lexical decision. PMID- 15358794 TI - Sentence production by young and older adults in controlled contexts. AB - In this experiment we compared young and older adults' abilities to produce complex sentences under controlled conditions. We asked participants to memorize sentence stems differing in syntactical complexity and then to produce a complete sentence using the stem. The length, complexity, and content of young adults' responses varied with the syntactical complexity of the stems, whereas older adults' responses did not. These results suggest that working memory processing limitations impose a "ceiling" on older adults' production of complex sentences, limiting their length, complexity, and content. PMID- 15358795 TI - Ipsilateral coordination deficits and central processing requirements associated with coordination as a function of aging. AB - Young and elderly participants performed concurrent ipsilateral hand-foot movements either isodirectionally or nonisodirectionally. We determined performance by measuring the maximal cycling frequency at which the coordination pattern could be performed successfully (CF(max)). We also determined attentional costs by means of a dual-task paradigm. Findings revealed that CF(max) was significantly lower in the elderly than in the young participants for the nonisodirectional mode, whereas we observed no differences for the isodirectional mode. Under dual-task conditions, coordination deteriorated in the elderly group only. However, when we equated levels of task difficulty, differences between the groups disappeared. Furthermore, attentional costs did not differ between isodirectional and nonisodirectional movements. This indicates that age-related coordination deficits were not primarily evoked by reduced attentional resources or control in elderly persons. PMID- 15358796 TI - Honeymoons and joint lunches: effects of retirement and spouse's employment on depressive symptoms. AB - With hypotheses derived from a life course perspective in conjunction with life event stress and role theories, we examine whether a spouse's employment and length of retirement affect a person's postretirement depressive symptoms and whether such effects differ by gender. Analyses use pooled data from Waves 1-4 of the Health and Retirement Survey, using a subsample of married individuals who either remained continuously employed over time or completely retired since the Wave 1 interviews (N = 2,695). Recently retired men seem to be negatively affected by their spouses' continuous employment when compared with men whose wives were continuously not employed. In contrast, spouses' joint retirement has a beneficial influence on both recently retired and longer-retired men. However, for recently retired men, the positive effect of wives' retirement seems to be contingent on spouses' enjoyment of joint activities. Among women, effects of spouses' employment occur only among very recently retired wives (0-6 months). These wives report more depressive symptoms if their spouses were already nonemployed prior to wives' retirement. These results demonstrate the complexity of retirement adaptation processes and suggest that marital context plays an important role in retirement well-being. PMID- 15358797 TI - Subjective usefulness and 6-year mortality risks among elderly persons in Japan. AB - Several studies have demonstrated that the loss of a sense of one's own self worth has an influence on the health status, psychological functioning, and quality of life in the aged. We used longitudinal data from 784 elderly residents (mean age = 73 years) to examine the relationship between subjective usefulness and 6-year mortality. We hypothesized that subjective usefulness is an important indicator of quality of life among the elderly population, and subsequently of mortality. We elicited information on subjective usefulness at baseline by using a self-administered questionnaire. Results of longitudinal analyses showed that subjective usefulness may be significantly associated with self-rated health and subsequent mortality. This indicates that subjective usefulness plays an important role in enhancing survival in the elderly population. PMID- 15358798 TI - The interpretation of social comparison and its relation to life satisfaction among elderly people: does frailty make a difference? AB - We examined the interpretation of upward and downward social comparison and its effect on life satisfaction in a questionnaire study among 444 community-dwelling elderly persons with different levels of frailty. As we expected, elderly persons with higher levels of frailty were less inclined to contrast and more inclined to identify themselves with a downward comparison target. Furthermore, they were more inclined to contrast themselves with an upward comparison target, but contrary to our expectations, they were also more inclined to identify with this target. Upward identification and downward contrast related positively, whereas upward contrast and downward identification related negatively to life satisfaction. These effects existed independently of the negative effect of frailty on life satisfaction. PMID- 15358799 TI - Volunteerism and socioemotional selectivity in later life. AB - OBJECTIVE: The goal of this work was to assess the applicability of socioemotional selectivity theory to the realm of volunteerism by analyzing data drawn from the September 2002 Current Population Survey Volunteer Supplement. METHODS: Total number of organizations volunteered for and total number of hours engaged in volunteer activities were utilized to obtain measures of volunteer hours per organization and volunteer hours in the main organization to determine whether a selective process could be observed. Descriptive statistics on age patterns were followed by a series of curve estimations to identify the best fitting curves. RESULT: Logistic age patterns of slowly increasing then relatively stable volunteer activity suggest that socioemotional selectivity processes are operative in the realm of voluntary activities. DISCUSSION: Socioemotional selectivity theory is applicable to voluntary activities. PMID- 15358800 TI - Formal volunteering as a protective factor for older adults' psychological well being. AB - OBJECTIVE: Guided by interactional role theory and employing a resilience framework, this study aimed to investigate whether formal volunteering protects older adults with more role-identity absences in major life domains (partner, employment, and parental) from poorer psychological well-being. METHODS: We used data from 373 participants, aged 65-74, in the 1995 National Survey of Midlife Development in the U.S. (MIDUS). Multivariate regression models estimated the effects of major role-identity absences, formal volunteering, and the interaction between major role-identity absences and volunteering on respondents' negative affect, positive affect, and purpose in life. RESULT: Participants with a greater number of major role-identity absences reported more negative affect, less positive affect, and less purpose in life. Being a formal volunteer was associated with more positive affect and moderated the negative effect of having more major role-identity absences on respondents' feelings of purpose in life. DISCUSSION: Consistent with previous studies, findings indicate that having more role-identity absences constitutes a risk factor for poorer psychological well being. Results further demonstrate that being a formal volunteer can protect older adults with a greater number of major role-identity absences from decreased levels of purpose in life. The findings suggest that associations between volunteering and psychological well-being might be contingent upon the volunteer's role-identity status and the dimension of psychological well-being examined. PMID- 15358801 TI - Household disbandment in later life. AB - OBJECTIVE: This study described activities that older people undertake to reduce the volume of their possessions in the course of a residential move to smaller quarters, a process with practical, cognitive, emotional, and social dimensions. METHODS: Qualitative interviews were conducted with members of 30 households who had moved in the prior year. The disbandment period, typically lasting about 2 months, was a particular focus of the interview. RESULT: The interviews suggested nine reasons why people had accumulated and kept things, which now became problematic for the impending move. The initial steps of disbandment entailed decisions about major furniture and meaningful gifts to family and friends, followed by evaluation of the remaining belongings for retention, sale, further gifts, donation, or discard. Things not divested by one means were reassigned to another strategy. People took pleasure in dispositions that saw their things used, cared for, and valued as they had done, thus fulfilling a responsibility to their belongings. DISCUSSION: Disbandment is an acute episode of a more general, lifelong process of possession management. It is an encounter with things that are meaningful to the self, but as it unfolds, it also makes new meaning for things. PMID- 15358802 TI - Gift wrapping ourselves: the final gift exchange. AB - OBJECTIVE: This qualitative study examines the "final gift exchange" process by which older adults give cherished possessions in return for lasting appreciation. METHODS: We interviewed 54 middle- and upper-middle-class people (39 women) aged 50-90 who had to dispose of personal objects when moving to smaller residences. RESULT: We used Goffman's spiraling strategy in our analysis of people's reports of disposing of personal possessions. We identified three salient dimensions (family, economy, and self) of this process and created a heuristic describing eight ideal-type gift exchange scenarios by categorizing objects as valued or not valued by family and the economy as well as being an important aspect of the gift giver's material self. DISCUSSION: By applying the heuristic, we observed that a lack of shared definitions of the meaning and value of objects created dilemmas in disposing of personal objects, particularly those connected to a person's material self. We also offer suggestions for making the process go more smoothly, such as inquiring about the preferences of others and telling stories associated with objects to create shared definitions of the objects' significance to the giver and/or to family history. PMID- 15358803 TI - Problems with vision associated with limitations or avoidance of driving in older populations. AB - OBJECTIVES: This report examines the role of (a) disease processes affecting vision, (b) reported troubles with vision, (c) physical symptoms affecting the eyes, and (d) objective measures in reported driving limitation due to problems with eyesight among older drivers. METHODS: Data for this study (N = 1,840) were obtained from participants in a community-based study of aging and physical performance in people age 55 or older in the city of Sonoma, California. Each of 16 visual conditions was assessed for impact on reported driving limitation due to eyesight by calculating a "risk" ratio. Then, prevalence of the condition was combined with the ratio to generate an attributable risk for that condition for vision-related limitations in driving. RESULTS: Each condition was significantly associated with reported limitations in driving due to eyesight. "Avoiding physical activity due to vision" (ratio = 3.4) and "trouble seeing steps up/down stairs" (ratio = 2.9) had the strongest association. However, "glasses/contacts required for driving" and "trouble with glare from sun/lights" had the highest attributable risks (35.8 and 29.4). DISCUSSION: The risk ratio is relevant for evaluating individuals; the attributable risk is relevant to planning countermeasures in populations. Addressing specific problems related to vision should substantially reduce driving limitations due to eyesight. PMID- 15358804 TI - Stressors arising in highly valued roles, meaning in life, and the physical health status of older adults. AB - OBJECTIVE: The purpose of this study is to see if stressors arising in highly valued roles affect health by eroding an older person's sense of meaning in life. A second goal is to assess whether emotional support provided by social network members helps offset the deleterious effects of stress on meaning and health. METHODS: Interviews were conducted with a nationwide sample of older people. Survey items were administered to assess stressors in highly valued roles, meaning in life, and self-rated health. RESULT: The findings suggest that life events arising in roles that are valued highly are associated with less favorable health ratings. Moreover, the data reveal that stressors in highly valued roles affect health primarily by eroding an older person's sense of meaning in life. Finally, the results indicate that emotional support helps older people cope more effectively with stress by restoring their sense of meaning in life. DISCUSSION: Researchers have devised a number of interventions to improve the health of older people by enhancing the functioning of their social networks. These interventions have met with mixed success. The findings from the current study suggest that focusing on meaning in life may help researchers devise more effective interventions. PMID- 15358805 TI - A prototype antibody microarray platform to monitor changes in protein tyrosine phosphorylation. AB - Reversible protein phosphorylation is a key regulatory process in all living cells. Deregulation of modification control mechanisms, especially in the case of tyrosine, may lead to malignant transformation and disease. Phosphotyrosine (p Tyr) accounts for only 0.05% of the total cellular phospho-amino acid content, yet plays an unusually prominent role in eukaryotic signaling, development, and growth. Tracking temporal and positional p-Tyr changes across the cellular proteome, i.e. tyrosine phosphoproteomics, is therefore tremendously valuable. Here, we describe and evaluate a prototype antibody (Ab) microarray platform to monitor changes in protein Tyr phosphorylation. Availability permitting, a virtually unlimited number of Abs, each recognizing a specific cellular protein, may be arrayed on a chip, incubated with total cell or tissue extracts or with biological fluids, and then probed with a fluorescently labeled p-Tyr-specific monoclonal Ab, PY-KD1, specifically generated for this assay as part of the current study. The optimized protocol allowed detection of changes in the Tyr phosphorylation state of selected proteins using submicrogram to low nanogram of total protein extract, amounts that may conceivably be obtained from a thousand to a hundred thousand cells, or less, depending on the cell or tissue type. The assay platform was evaluated by assessing changes in a rationally selected subset of the Tyr phosphoproteome of Bcr-Abl-expressing cells treated with a specific inhibitor, Gleevec, and of epidermal growth factor (EGF)-treated HeLa cells. The results, ratiometric rather than strictly quantitative in nature, conformed with previous identifications of several Bcr-Abl and EGF receptor targets, and associated proteins, as detected by exhaustive mass spectrometric analyses. The Ab microarray method described here offers advantages of low sample and reagent consumption, scalability, detection multiplexing, and potential compatibility with microfluidic devices and automation. The system may hold particular promise for dissecting signaling pathways, molecular classification of tumors, and profiling of novel target-cancer drugs. PMID- 15358806 TI - Effect of timing of iron supplementation on maternal and neonatal growth and iron status of iron-deficient pregnant rats. AB - We have previously shown that maternal iron (Fe) deficiency not only reduces fetal size, but also increases blood pressure in the offspring when they are adults. In this paper we examine whether there are critical periods when supplementation reverses or fails to reverse the effect both on size and on expression of genes of Fe metabolism. We made dams Fe deficient, mated them and provided supplements of Fe in the diet from the beginning of gestation (0.5 days), from 7.5 days or from 14.5 days. Within 12 h of birth, dams and neonates were killed and tissues taken and examined. Fe deficiency throughout pregnancy reduces neonatal size. Supplementation from the beginning of the first, second or third week all reduced the effect. Maternal haematocrit was restored to normal levels only in animals given supplements for at least 2 weeks. In contrast, the neonates' Fe levels were normal in all supplemented groups. These results were mirrored in liver Fe levels and in transferrin receptor mRNA. Iron-responsive element (IRE)-regulated divalent metal transporter 1 (DMT1) increased in maternal and neonatal liver. Non-IRE-regulated DMT1 levels did not change in the maternal liver, but decreased in the neonatal liver. H and L ferritin mRNA levels also showed different patterns in the mother and her offspring. Finally, the neonatal size correlated with maternal Fe stores, and not with those of the fetus. The data demonstrate that Fe supplementation during pregnancy is most effective when given early, rather than later, in gestation. PMID- 15358807 TI - Postnatal maturation of mossy fibre excitatory transmission in mouse CA3 pyramidal cells: a potential role for kainate receptors. AB - Kainate receptors (KARs) are abundantly expressed in the central nervous system at a period of intense synaptogenesis and might participate in the maturation of neural networks. We have described the postnatal development of mossy fibre excitatory synaptic transmission in CA3 pyramidal cells and we have explored the potential role of KARs in synaptic maturation. In CA3 pyramidal cells, mossy fibre stimulation evokes EPSCs as early as postnatal day 3 (P3). At this early stage, mossy fibre (MF)-EPSCs are fully blocked by GYKI 53655, an AMPA receptor (AMPAR) antagonist. A postsynaptic KAR component can only be detected from P6. Thus, AMPAR-EPSCs precede KAR-EPSCs during postnatal maturation at this synapse. All MF-EPSCs display a KAR component after P10. A key issue of the present work is that between P6 and P9, the presence of a postsynaptic KAR component tightly coincides with AMPAR-mediated EPSCs of large amplitude, and with the onset of low frequency facilitation (from 0.1 Hz to 1 Hz), a presynaptic form of short-term synaptic plasticity. In addition, mice lacking functional KARs throughout postnatal development display MF-EPSCs of significantly smaller amplitude at stages of maturation where synaptic KARs are normally present, due to both pre- and postsynaptic impairment of synaptic transmission. These data suggest a role for KARs in the maturation of mossy fibre synapses. PMID- 15358808 TI - Influence of nitric oxide synthase inhibition on pulmonary O2 uptake kinetics during supra-maximal exercise in humans. AB - We have recently reported that inhibition of nitric oxide synthase (NOS) with N(G)-nitro-L-arginine methyl ester (L-NAME) accelerates the 'phase II' pulmonary O2 uptake (VO2) kinetics following the onset of moderate and heavy intensity submaximal exercise in humans. These data suggest that the influence of nitric oxide (NO) on mitochondrial function is an important factor in the inertia to aerobic respiration that is evident in the transition from a lower to a higher metabolic rate. The purpose of the present study was to investigate the influence of L-NAME on pulmonary VO2 kinetics following the onset of supra-maximal exercise, where it has been suggested that O2 availability represents an additional limitation to VO2 kinetics. Seven healthy young men volunteered to participate in this study. Following an incremental cycle ergometer test for the determination of VO2max, the subjects returned on two occasions to perform a 'step' exercise test from a baseline of unloaded cycling to a work rate calculated to require 105% VO2max, preceded either by systemic infusion of L-NAME (4 mg kg(-1) in 50 ml saline) or 50 ml saline as a control (Con). Pulmonary gas exchange was measured on a breath-by-breath basis throughout the exercise tests. The duration of 'phase I' was greater with L-NAME (Con: 14.0 +/- 2.1 versus L NAME: 16.0 +/- 1.6 s; P = 0.03), suggestive of a slower cardiovascular adaptation following the onset of exercise. However, the phase II VO2 time constant was reduced by 44% with L-NAME (Con: 36.3 +/- 17.3 versus L-NAME: 20.4 +/- 8.3 s; P = 0.01). The accumulation of blood lactate during exercise was also reduced with L NAME (Con: 4.0 +/- 1.1 versus L-NAME: 2.7 +/- 2.1 mM; P = 0.04). These data indicate that skeletal muscle NO production represents an important limitation to the acceleration of oxidative metabolism following the onset of supra-maximal exercise in humans. PMID- 15358809 TI - The effect of transcranial magnetic stimulation and peripheral nerve stimulation on corticomuscular coherence in humans. AB - Cortex and muscle show coupled oscillations in the 15-35 Hz frequency band during voluntary movements. To obtain evidence of the neuronal network responsible for this rhythmicity we investigated the effect of transcranial magnetic stimulation (TMS) and peripheral nerve stimulation on the coupling between eletcroencephalographic (EEG) activity recorded from the scalp over the motor cortex and electromyographic (EMG) activity recorded from the tibialis anterior (TA) muscle in 15 healthy human subjects. TMS over the leg area at intensities between 0.95 and 1.1 x threshold for a motor evoked potential (MEP) in the TA increased corticomuscular coherence in the 15-35 Hz frequency band. This effect lasted on average for 300 ms, but could last up to 600-800 ms in some subjects. Stimulation of motor nerves from the ankle muscles suppressed corticomuscular coherence in the 15-35 Hz frequency range between leg area EEG and TA EMG for a period up to 600-800 ms. In addition, increased coherence around 10 Hz was observed for a period up to 250 ms after the stimulation. Stimulation of motor nerves in the arm and motor nerves from the ankle muscles in the other leg had no effect. The findings indicate that TMS has direct access to the neuronal circuitry in the motor cortex, which generates the corticomuscular coherence. This effect was caused either by direct activation of corticospinal cells or by activation of local neuronal circuitries in the motor cortex. The effects of peripheral nerve stimulation suggest that an alternative rhythm generator may entrain the cortical cells into a lower 10 Hz rhythm and disrupt the 15-35 Hz rhythm. PMID- 15358810 TI - Calcium channels activated by endothelin-1 in human trophoblast. AB - Ca2+ transfer across the syncytiotrophoblast (ST) of the human placenta is essential for normal fetal development. However, the nature of Ca2+ conductance in the ST and the mechanisms by which it is regulated are poorly understood. With the major signal transduction pathway of endothelin-1 (ET1) acting via phospholipase C (PLC) and Ca2+, we used ET1 to analyse the nature of Ca2+ channels on cultured trophoblastic cells by means of cytofluorimetric analysis using the ratiometric Ca2+ indicator Indo-1. Results indicate that ET1 (10(-7) M) stimulates a biphasic (transient and sustained) increase in [Ca2+]i in trophoblastic cells. This response is mediated by the endothelin receptor B (ETB) coupled to PLC, since treatment with BQ788 (10(-6) M) or U73122 (2 microM) totally abolished the response. Persistence of the rapid transient rise in [Ca2+]i in Ca2+-free extracellular medium confirms the release of Ca2+ from intracellular stores in response to ET1 stimulation. Furthermore, abolition of the sustained increase in [Ca2+]i in Ca2+-free extracellular medium argues in favour of the entry of Ca2+ during the plateau phase. Abolition of this plateau phase by Ni2+ (1 mM) in the presence of extracellular Ca2+ confirmed the existence of an ET1-induced Ca2+ entry. No evidence for the presence of voltage operated channels was demonstrated during ET1 action since nifedipine (10(-6) M) did not reduce the Ca2+ response and depolarization with a hyper-potassium solution had no effect. Pharmacological studies using the imidazole derivatives SK&F96365 (30 microM) and LOE 908 (10 microM) partially inhibited the ET1-evoked Ca2+ response, thus providing evidence for the presence of both store-operated Ca2+ channels and non-selective cationic channels in the human ST. PMID- 15358811 TI - NG2-positive cells in the mouse white and grey matter display distinct physiological properties. AB - Cells that express the NG2 proteoglycan are the largest proliferative progenitor population in the postnatal central nervous system (CNS). Although this entire population has long been considered to be oligodendrocyte progenitors, numerous NG2(+) cells are present in the cerebral cortex, where relatively little myelination occurs, and also persist long after myelination is complete in the CNS. Several studies have alluded to the presence of distinct NG2(+) cell subtypes based on marker expression, but no experimentally derived hypotheses about the physiological role of these subtypes has been proposed. In the current study, whole-cell patch-clamp data from acutely isolated slices demonstrate that subcortical white matter and cortical NG2(+) cells display distinct membrane properties in addition to possessing differing K(+)- and Na(+)-channel expression profiles. A striking observation is that a subpopulation of cortical, but not white matter NG2(+) cells, elicit depolarization-induced spikes that are akin to immature action potentials. Our data demonstrate that a population of cortical NG2(+) cells display physiological properties that differ from their white matter counterparts. PMID- 15358812 TI - The combi-targeting concept: dissection of the binary mechanism of action of the combi-triazene SMA41 in vitro and antitumor activity in vivo. AB - We have previously reported the synthesis of SMA41, a unimolecular combination of an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) of the quinazoline class and a DNA-damaging monomethyltriazene termed "combimolecule". Hydrolysis of 1-[4-(m-tolylamino)-6-quinazolinyl]-3-methyltriazene (SMA41) gives rise to an intact TKI [6-amino-4-(3-methylanilino)quinazoline; SMA52] capable of inhibiting epidermal growth factor (EGF)-induced EGFR autophosphorylation and a DNA-targeting methyldiazonium species. Herein, we showed that SMA41 blocked EGF induced EGFR autophosphorylation by an irreversible mechanism, suggesting that it may covalently damage the receptor in these cells. More importantly, this was associated with significant inhibition of mitogen-activated protein kinase activation in A431 cells. In cells treated with [14C]SMA41, radio-high performance liquid chromatography detection of both N7- and O6-methylguanine revealed an almost complete repair of the O6-methylguanine lesions and a greater tolerance of the N7-methylguanine adducts 24 h post-treatment. In contrast to temozolomide (a cyclic triazene used in the clinic) and the reversible inhibitor SMA52, SMA41 induced significant cell cycle arrest in S, G2, and M phases 24 h after a 2-h drug exposure. Furthermore, in vivo studies demonstrated that SMA41 was well tolerated. At 200 mg/kg, it showed approximately 2-fold greater antiproliferative activity than SMA52 in A431 cells implanted in immunocompromised SCID mice. These results suggest that the binary targeting properties of SMA41 are associated with a binary cascade of events in the cells that seem to culminate into significant growth inhibition in vitro and in vivo. PMID- 15358813 TI - Prolyl endopeptidase-mediated destruction of T cell epitopes in whole gluten: chemical and immunological characterization. AB - Celiac Sprue is a widely prevalent immune disease of the small intestine induced by dietary gluten intake in genetically susceptible individuals. It has been suggested that prolyl endopeptidases (PEPs) may be useful catalysts for gluten detoxification. We have investigated this hypothesis using food-grade gluten as the target antigen, and a combination of mass spectrometry and patient-derived T cells as quantitative assay systems. Spectrometric characterization of physiologically proteolyzed gluten revealed a number of 10 to 50 residue peptides containing known T cell epitopes involved in Celiac Sprue pathogenesis. Several of these peptides were multivalent, suggesting they may be potent triggers of the inflammatory response to gluten in celiac patients. Treatment of proteolyzed gluten with recombinant bacterial PEP decreased the number of potentially immunostimulatory peptides. Substantially reduced immunogenicity was also quantified in 12 of 14 intestinal polyclonal T cell lines from celiac patients. Kinetic investigations using eight T cell clones showed rapid destruction of alpha-gliadin epitopes, but less complete processing of gamma-gliadin epitopes. Given the difficulty associated with a strict lifelong gluten-exclusion diet, the ability of a single enzyme to greatly reduce the antigenic burden of grocery store gluten reinforces the case for developing oral peptidase therapy against Celiac Sprue. PMID- 15358814 TI - Chronic administration of statins alters multiple gene expression patterns in mouse cerebral cortex. AB - Statins have been reported to lower the risk of developing Alzheimer's disease; however, the mechanism of this potentially important neuroprotective action is not understood. Lowering cholesterol levels does not appear to be the primary mechanism. Statins have pleiotropic effects in addition to lowering cholesterol, and statins may act on several different pathways involving distinct gene expression patterns that would be difficult to determine by focusing on a few genes or their products in a single study. In addition, gene expression patterns may be specific to a particular statin. To understand the molecular targets of statins in brain, DNA microarrays were used to identify gene expression patterns in the cerebral cortex of mice chronically treated with lovastatin, pravastatin, and simvastatin. Furthermore, brain statin levels were determined using liquid chromatography/tandem mass spectrometry. These studies revealed 15 genes involved in cell growth and signaling and trafficking that were similarly changed by all three statins. Overall, simvastatin had the greatest influence on expression as demonstrated by its ability to modify the expression of 23 genes in addition to those changed by all three drugs. Of particular interest was the expression of genes associated with apoptotic pathways that were altered by simvastatin. Reverse transcription-polymerase chain reaction experiments confirmed the microarray findings. All three drugs were detected in the cerebral cortex, and acute experiments revealed that statins are relatively rapidly removed from the brain. These results provide new insight into possible mechanisms for the potential efficacy of statins in reducing the risk of Alzheimer's disease and lay the foundation for future studies. PMID- 15358815 TI - Sorting a Staphylococcus aureus phage display library against ex vivo biomaterial. AB - A phage display library made from Staphylococcus aureus DNA was sorted against a central venous catheter (CVC) that had been removed from a patient 2 days after insertion. After the first panning, approximately 50% of the clones encoded proteins known to interact with mammalian proteins. After the second and third pannings, fibrinogen-binding and beta2-glycoprotein I (beta2-GPI)-binding phage particles were clearly dominating. Proteins adsorbed to different CVCs were investigated using specific antibodies. Among the proteins probed for, fibrinogen was most abundant, but, interestingly, beta2-GPI was also detected on all tested CVCs. PMID- 15358816 TI - Factors affecting the escape of Francisella tularensis from the phagolysosome. AB - The highly virulent bacterium Francisella tularensis is well adapted to the intracellular habitat but the mechanisms behind its intracellular survival have been elusive. Recently, it was shown that the bacterium is capable of escaping from the phagosome of human and mouse monocytic cells. Here it is shown that this escape is affected by gamma interferon (IFN-gamma) treatment of mouse peritoneal exudate cells since in treated cells the proportion that escaped was significantly lower (80%) than in untreated cells (97%) as determined by transmission electron microscopy. By contrast, < 1% of mutant bacteria lacking expression of a 23 kDa protein denoted IglC were able to escape from the phagosome. Infection with the DeltaiglC strain complemented with the iglC gene resulted in 60% of the bacteria escaping from the phagosome. Whereas IFN-gamma treatment conferred a static effect on intracellular wild-type bacteria, the treatment had a bactericidal effect on the DeltaiglC strain. The results show that the activation status of infected cells affects the escape of F. tularensis from the phagosome. An even more profound effect on this escape is related to expression of IglC by F. tularensis. Its absence rendered the mutant bacteria incapable of escaping from the phagosome and of multiplying intracellularly. PMID- 15358817 TI - The distribution of, and antibody response to, the core lipopolysaccharide region of Escherichia coli isolated from the faeces of healthy humans and cattle. AB - There are five different core types of Escherichia coli lipopolysaccharide (LPS), and enterohaemorrhagic E. coli tend to have the R3 core type. It has been hypothesized that increased carriage of bacteria with a specific core type will induce higher levels of antibodies and protect against disease caused by bacteria carrying that specific LPS core. Approximately 320 isolates of E. coli, half from healthy human faeces and half from healthy bovine faeces have been core typed both by core-specific monoclonal antibodies, and by PCR for genes encoding the enzymes responsible for the biosynthesis of the specific core structures. Results showed that E. coli possessing R1 core LPS were most frequently detected in both human and cattle populations (63 and 49%, respectively). Compared to the human isolates a significantly higher level of bacteria with R3 core LPS was detected among the bovine commensal E. coli (11% compared to 4%; P < 0.05). Antibody levels to each of the specific core types were measured in serum samples from healthy humans (n = 91) and healthy cattle (n = 39). In each population the highest level of antibody detected was reactive to the R4 core. In cattle the level of anti-R3 core antibody was significantly higher than the level of anti R1, -R2 and -K12 antibodies (P < 0.01). In summary there was a greater proportion of E. coli with R3 core type in cattle, together with a corresponding higher anti R3 antibody level. This suggests that cattle may have greater immunity to E. coli strains with an LPS of R3 core type. PMID- 15358818 TI - Analysis of gene expression profile in gastric cancer cells stimulated with Helicobacter pylori isogenic strains. AB - To understand the biological processes within host cells induced by VacA, isogenic strains of Helicobacter pylori (NCTC 11638 or 11638-DeltavacA) were used to stimulate gastric cancer cells SGC7901, and differentially expressed genes in host cells were identified using cDNA microarray technology. More than 300 genes were found to alter their mRNA expression at different time points, among which 68 were related to the cytoskeleton, 87 were associated with cell cycle, cell death and proliferation, IL8 expression was also found to be up-regulated. Cells co-cultured with broth-culture supernatant (BCS) of NCTC 11638 showed more alteration in microtubule cytoskeleton morphology, as observed by laser scanning confocal microscopy, and a lower apoptosis rate, detected by flow cytometry, compared with those co-cultured with BCS of 11638-DeltavacA. The supernatants of cells co-cultured with NCTC 11638 showed significantly higher IL8 expression than those co-cultured with 11638-DeltavacA. It is concluded that VacA disrupts cytoskeletal architecture by influencing the expression of cytoskeleton associated genes. VacA breaks the balance between cell proliferation and cell death by inducing the maladjustment of genes related to cell cycle. VacA is also able to induce the inflammatory response. PMID- 15358819 TI - Expression of leptospiral immunoglobulin-like protein by Leptospira interrogans and evaluation of its diagnostic potential in a kinetic ELISA. AB - The search for novel antigens suitable for improved vaccines and diagnostic reagents against leptospirosis led to the identification of LigA and LigB. LigA and LigB expression were not detectable at the translation level but were detectable at the transcription level in leptospires grown in vitro. Lig genes were present in pathogenic serovars of Leptospira, but not in non-pathogenic Leptospira biflexa. The conserved and variable regions of LigA and LigB (Con, VarA and VarB) were cloned, expressed and purified as GST-fusion proteins. Purified recombinant LigA and LigB were evaluated for their diagnostic potential in a kinetic ELISA (KELA) using sera from vaccinated and microscopic agglutination test (MAT)-positive dogs. Sera from vaccinated dogs showed reactivity to whole-cell antigens of leptospires but did not show reactivity in the KELA assay with recombinant antigens, suggesting a lack of antibodies to Lig proteins in the vaccinated animals. The diagnostic potential of recombinant Lig antigens in the KELA assay was evaluated by using 67 serum samples with MAT > or =1600, which showed reactivity of 76, 41 and 35% to rConA, rVarA and rVarB, respectively. These findings suggest that recombinant antigen to the conserved region of LigA and LigB can differentiate between vaccinated and naturally infected animals. PMID- 15358820 TI - Assessment of Chlamydia trachomatis infection in asymptomatic male partners of infertile couples. AB - Three specimens from 111 asymptomatic male partners of infertile couples attending the Department of Urology in Amiens, France, were examined by the PCR COBAS AMPLICOR test (Roche Molecular Diagnostics) for the presence of Chlamydia trachomatis. The specimens analysed were: first void urine (FVU), urine obtained after prostatic massage (UPM) and semen specimens. Serum from each patient was also obtained and analysed for the presence of IgG and IgA chlamydial antibodies by in-house microimmunofluorescence (MIF) and pELISA. C. trachomatis was detected by PCR in 5.4% of FVU samples, 2.7% of semen specimens and in 0.9% of UPM samples. Two treatments for processing the samples (storage at -70 degrees C and heating to 95 degrees C) were routinely used before initial testing to reduce the effects of inhibitors of PCR. Despite these precautions, the PCR method revealed the presence of inhibitors in 7.3% of semen specimens and 3.6% of FVU samples. C. trachomatis was detected by PCR COBAS AMPLICOR in seven of 111 patients (6.3%) and by serology in five of 111 patients (4.5%). The detection of C. trachomatis in FVU, UPM and semen specimens can serve as a marker for the presence of this organism in the genital tract, and can be used as a reliable way of detecting asymptomatic carriers of infection. PMID- 15358821 TI - Macrorestriction analysis of Streptococcus agalactiae (group B Streptococcus) isolates from Malaysia. AB - Streptococcus agalactiae or group B streptococci (GBS) often colonize the gastrointestinal and urogenital tracts of women, who may transmit these organisms to their offspring during the birth process. Using PFGE analysis, the genetic diversity of GBS was studied for strains isolated from pregnant women and their newborn infants in a teaching hospital. A total of 48 different PFGE profiles were obtained from 123 strains, with one profile (S1) appearing to be predominant among both groups studied. There was good overall correlation between the profiles obtained for strains from mother-infant pairs and for strains isolated from different body sites in the same individual. Occasional discrepancies seen in related body sites and among mother-infant pairs suggest concurrent carriage of different strains in the same individual as well as the possibility of an environmental source of organism for the neonate. The overall results demonstrated that many variants of GBS strains occur in Malaysia. PMID- 15358822 TI - Characterizing uncommon Burkholderia cepacia complex isolates from an outbreak in a haemodialysis unit. AB - An outbreak of bacteraemia in a haemodialysis unit where 65 episodes of infection involved 35 outpatients is reported. Burkholderia cepacia complex was the agent most frequently recovered from blood. Thirty-three environmental and clinical isolates of B. cepacia complex were characterized by whole-cell protein electrophoresis and recA-RFLP profile. Fourteen isolates were genomovar I and 16 isolates were not classifiable by their recA-RFLP pattern. Ribotyping, random amplification of polymorphic DNA (RAPD) and integron profile were used to explore the clonality of the isolates, and revealed multiple strain genotypes. Four ribotypes and RAPD types and three integron patterns were identified. The water supply was identified as the source of the outbreak, and inappropriate cleaning and a leak in the reverse osmosis tubing connection were the probable causes of contamination. B. cepacia complex was still recovered from blood of patients even after apparently adequate measures were taken and water quality standards were met, suggesting that higher standards for water quality should be adopted in haemodialysis units. The genomovars recovered here were distinct from those commonly reported for cystic fibrosis isolates. PMID- 15358823 TI - Post-mortem culture of Balamuthia mandrillaris from the brain and cerebrospinal fluid of a case of granulomatous amoebic meningoencephalitis, using human brain microvascular endothelial cells. AB - The first isolation in the UK of Balamuthia mandrillaris amoebae from a fatal case of granulomatous amoebic meningoencephalitis is reported. Using primary cultures of human brain microvascular endothelial cells (HBMECs), amoebae were isolated from the brain and cerebrospinal fluid (CSF). The cultures showed a cytopathic effect at 20-28 days, but morphologically identifiable B. mandrillaris amoebae were seen in cleared plaques in subcultures at 45 days. The identification of the organism was later confirmed using PCR on Chelex-treated extracts. Serum taken while the patient was still alive reacted strongly with slide antigen prepared from cultures of the post-mortem isolate, and also with those from a baboon B. mandrillaris strain at 1:10,000 in indirect immunofluorescence, but with Acanthamoeba castellanii (Neff) at 1:160, supporting B. mandrillaris to be the causative agent. If the presence of amoebae in the post mortem CSF reflects the condition in life, PCR studies on CSF and on biopsies of cutaneous lesions may also be a valuable tool. The role of HBMECs in understanding the interactions of B. mandrillaris with the blood-brain barrier is discussed. PMID- 15358824 TI - Efficacy of antiseptics and disinfectants on clinical and environmental yeast isolates in planktonic and biofilm conditions. AB - The aim of this study was to evaluate the efficacy of five antiseptics, three surface disinfectants and UV radiation against a wide range of clinical and environmental yeast isolates. Their efficacy against pure cultures, yeast mixtures and biofilms (prepared by culturing yeasts in Sabouraud broth containing a final concentration of 8% glucose) was tested. Three clinical isolates of Candida albicans, Cryptococcus neoformans and Rhodotorula rubra, and two environmental isolates of Candida albicans and Cryptococcus uniguttulatus were selected. For seven out of eight biocides tested (Betadine, Dermacide, Chlorhexidine, Dosisepsine, hydrogen peroxide, sodium hypochlorite, 70% alcohol, 0.5% Ecodiol) and for UV radiation, susceptibility did not differ according to genus, species or origin. Hydrogen peroxide, 0.25% Ecodiol and UV radiation were ineffective against the five isolates tested. On pure planktonic cultures, and, to a lesser extent, on free-living yeast mixtures, the other products were active and were then tested against biofilms: eight out of nine biocides were ineffective. Chlorhexidine at 0.5% was the only fungicidal agent on pure cultures, yeast mixtures and biofilms. The importance of the test method, including agent concentration, is discussed. PMID- 15358825 TI - In vitro activity of fluoroquinolone and the gyrA gene mutation in Helicobacter pylori strains isolated from children. AB - Resistance to antibiotics, especially clarithromycin, is the major cause of the failure to eradicate Helicobacter pylori. There are few studies in children concerning fluoroquinolone activity against H. pylori. Primary resistance to antibiotics including fluoroquinolones was studied in 55 H. pylori strains isolated from Japanese children. DNA sequences of the gyrA gene in fluoroquinolone-resistant strains were determined. Twelve strains (21.8%) were resistant to clarithromycin and three (5.5%) were resistant to both levofloxacin and ciprofloxacin. Out of 12 clarithromycin-resistant strains, 11 (91.7%) were susceptible to levofloxacin and ciprofloxacin. Sequence analysis in three fluoroquinolone-resistant strains showed point mutations of the gyrA gene at G271A, G271T and A272G, indicating mutations of the codon Asp91 in the fluoroquinolone-resistance-determining region of the DNA gyrase. The results suggest that fluoroquinolones should be considered as an option for second- or third-line H. pylori eradication therapy in children. PMID- 15358826 TI - Influence of subinhibitory concentrations of plant essential oils on the production of enterotoxins A and B and alpha-toxin by Staphylococcus aureus. AB - The data presented show the ability of subinhibitory concentrations of plant essential oils to influence the production of enterotoxins A and B and alpha toxin by Staphylococcus aureus. Subinhibitory concentrations of the oils of bay, clove, cinnamon, nutmeg and thyme had no significant effect on the overall quantity of extracellular protein produced. Haemolysis due to alpha-toxin was significantly reduced after culture with all five plant essential oils. This reduction was greatest with the oils of bay, cinnamon and clove. These three oils also significantly decreased the production of enterotoxin A; the oils of clove and cinnamon also significantly decreased the production of enterotoxin B. PMID- 15358827 TI - Real-time PCR investigation into the importance of Fusobacterium necrophorum as a cause of acute pharyngitis in general practice. AB - Fusobacterium necrophorum is recognized as the cause of a severe life-threatening illness characterized by bacteraemia with metastatic abscesses following an acute sore throat (Lemierre's disease). However, the importance of F. necrophorum as a cause of simple sore throat in the community is unknown. Using quantitative real time PCR with primers targeting the rpoB gene, 100 routine throat swabs collected from patients presenting to general practitioners with pharyngitis were analysed for the presence of F. necrophorum-specific DNA. The results were compared with those obtained from throat swabs collected from 100 healthy subjects. Ten clinical samples were positive for F. necrophorum DNA, identified as F. necrophorum subspecies funduliforme, using a haemagglutinin-related protein gene specific PCR assay. All the healthy controls were negative (two-tailed P value = 0.0015; Fisher exact test). These findings suggest that F. necrophorum may play a more important role as a cause of simple sore throat in the community than has been previously appreciated. PMID- 15358828 TI - Phenotypic and genotypic characterization of beta-D-glucuronidase-positive Shiga toxin-producing Escherichia coli O157:H7 isolates from deer. AB - Beta-glucuronidase-positive (GUD+) Shiga toxin-producing Escherichia coli (STEC) O157:H7 was isolated from both an asymptomatic woman and uncooked deer meat in her possession in Hokkaido, Japan. The phenotypic and genotypic characteristics of the two isolates were identical or closely related, indicating probable transmission of the deer isolate to the woman. Moreover, several other GUD+ STEC O157:H7 strains investigated belonged to the distinct atypical GUD+ STEC O157:H7 group that has been identified previously. This is the first report that deer can be a reservoir of GUD+ STEC O157:H7 in Japan. PMID- 15358829 TI - Community-acquired bacteraemia in a rural area: predominant bacterial species and antibiotic resistance. AB - The invasion of the bloodstream represents one of the most important sequelae of infection. This study was conducted over an 18-month period to determine the predominant bacterial agents of a community-acquired bacteraemia seen at health centres in a rural area of Jordan, and their antibiotic susceptibilities. Blood samples were collected and cultured from 215 patients who presented with fever and presumed diagnosis of a bacteraemia. Isolates were identified and tested for antibiotic susceptibility. The variables included the age and sex of the patients, aetiology, sources of the bacteraemia, risk factors, treatment and outcome. One hundred and twenty-six (58.6%) blood cultures were positive. Children less than 14 years old accounted for 34.9% of these, and 38% were from patients that were more than 50 years old. The most frequent aetiologic agents were Staphylococcus aureus, followed by Brucella melitensis and Streptococcus pneumoniae. A wide range of resistance to commonly used antimicrobial agents and multidrug resistance was documented in 44.4% of the isolates. The most frequent sources of the bacteraemia were urinary (15.9%), respiratory (14.3%), no source of the bacteraemia identified (primary bacteraemia) (13.5%), gastrointestinal (12.7%) and soft-tissue infection (7.9%). No identifiable risk factor for infection could be determined in 34% of the patients. The predominant pathogens identified and the relatively high prevalence of antibiotic resistance of the isolates are most probably due to the nature and lifestyle of this rural population and the use of empiric treatment. Characteristics permitting recognition of patients with such strains would aid infection control efforts in the community. PMID- 15358830 TI - A purulent pericarditis caused by Salmonella typhimurium. AB - A case of Salmonella typhimurium pericarditis is reported. The diagnosis was based on blood and pericardial effusion cultures. PMID- 15358831 TI - Biochemical characterization of Cdc6/Orc1 binding to the replication origin of the euryarchaeon Methanothermobacter thermoautotrophicus. AB - Archaeal cell division cycle protein 6 (Cdc6)/Origin Replication Complex subunit 1 (Orc1) proteins share sequence homology with eukaryotic DNA replication initiation factors but are also structurally similar to the bacterial initiator DnaA. To better understand whether Cdc6/Orc1 functions in an eukaryotic or bacterial-like manner, we have characterized the interaction of two Cdc6/Orc1 paralogs (mthCdc6-1 and mthCdc6-2) with the replication origin from Methanothermobacter thermoautotrophicus. We show that while both proteins display a low affinity for a small dsDNA of random sequence, mthCdc6-1 binds tightly to a short duplex containing a single copy of a 13 bp sequence that is repeated throughout the origin. Surprisingly, sequence comparisons show that this 13 bp sequence is a minimized version of the Origin Recognition Box element found in many euryarchaeotal origins. Analysis of mthCdc6-1 mutants demonstrates that the helix-turn-helix motif in the winged-helix domain mediates the interaction with this sequence. Association of both mthCdc6/Orc1 paralogs with the duplex containing the minimized Origin Recognition Box fits to an independent binding sites model, but their interaction with longer DNA ligands is cooperative. Together, our data provide the first detailed biophysical characterization of the association of an archaeal DNA replication initiator with its origin. Our observations also indicate that the origin-binding properties of Cdc6/Orc1 proteins closely resemble those of bacterial DnaA. PMID- 15358832 TI - SLBP is associated with histone mRNA on polyribosomes as a component of the histone mRNP. AB - The stem-loop binding protein (SLBP) binds the 3' end of histone mRNA and is present both in nucleus, and in the cytoplasm on the polyribosomes. SLBP participates in the processing of the histone pre-mRNA and in translation of the mature message. Histone mRNAs are rapidly degraded when cells are treated with inhibitors of DNA replication and are stabilized by inhibitors of translation, resulting in an increase in histone mRNA levels. Here, we show that SLBP is a component of the histone messenger ribonucleoprotein particle (mRNP). Histone mRNA from polyribosomes is immunoprecipitated with anti-SLBP. Most of the SLBP in cycloheximide-treated cells is present on polyribosomes as a result of continued synthesis and transport of the histone mRNP to the cytoplasm. When cells are treated with inhibitors of DNA replication, histone mRNAs are rapidly degraded but SLBP levels remain constant and SLBP is relocalized to the nucleus. SLBP remains active both in RNA binding and histone pre-mRNA processing when DNA replication is inhibited. PMID- 15358833 TI - Effective and robust plasmid topology analysis and the subsequent characterization of the plasmid isoforms thereby observed. AB - Within the biopharmaceutical industry, recombinant plasmid DNA is used both as a raw material (e.g. in lentiviral and AAV vector production) as well as an active ingredient (e.g. in DNA vaccines). Consequently, many analytical laboratories are routinely involved with plasmid DNA topoisoform qualitative analysis and quantification. In order to reliably determine plasmid topology, one must ensure that the methodology employed can reliably, precisely and accurately measure qualitatively and quantitatively all topological isoforms. Presented here are an anion-exchange high-performance liquid chromatography (AEC) and an agarose gel electrophoresis (AGE)-based method developed for this purpose. The strategies undertaken to overcome the respective typical problems of limited linear range of quantitation (for AGE) and isoform resolution (for AEC) are described. Also presented is a subsequent direct comparison (for assay precision/accuracy) of these two methods, as well as a package of species characterization [by chloroquine-AGE, enzymatic digestion, multi-angle laser light-scattering (MALLS) and electron microscopy] undertaken to confirm the identity of a minor supercoiled dimeric concatamer observed by both approaches. PMID- 15358834 TI - Gene expression profiling identifies a unique androgen-mediated inflammatory/immune signature and a PTEN (phosphatase and tensin homolog deleted on chromosome 10)-mediated apoptotic response specific to the rat ventral prostate. AB - Understanding androgen regulation of gene expression is critical for deciphering mechanisms responsible for the transition from androgen-responsive (AR) to androgen-independent (AI) prostate cancer (PCa). To identify genes differentially regulated by androgens in each prostate lobe, the rat castration model was used. Microarray analysis was performed to compare dorsolateral (DLP) and ventral prostate (VP) samples from sham-castrated, castrated, and testosterone replenished castrated rats. Our data demonstrate that, after castration, the VP and the DLP differed in the number of genes with altered expression (1496 in VP vs. 256 in DLP) and the nature of pathways modulated. Gene signatures related to apoptosis and immune response specific to the ventral prostate were identified. Microarray and RT-PCR analyses demonstrated the androgen repression of IGF binding protein-3 and -5, CCAAT-enhancer binding protein-delta, and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) genes, previously implicated in apoptosis. We show that PTEN protein was increased only in the luminal epithelial cells of the VP, suggesting that it may be a key mediator of VP apoptosis in the absence of androgens. The castration-induced immune/inflammatory gene cluster observed specifically in the VP included IL-15 and IL-18. Immunostaining of the VP, but not the DLP, showed an influx of T cells, macrophages, and mast cells, suggesting that these cells may be the source of the immune signature genes. Interestingly, IL-18 was localized mainly to the basal epithelial cells and the infiltrating macrophages in the regressing VP, whereas IL-15 was induced in the luminal epithelium. The VP castration model exhibits immune cell infiltration and loss of PTEN that is often observed in progressive PCa, thereby making this model useful for further delineation of androgen regulated gene expression with relevance to PCa. PMID- 15358835 TI - Orphan nuclear receptor small heterodimer partner represses hepatocyte nuclear factor 3/Foxa transactivation via inhibition of its DNA binding. AB - Small heterodimer partner (SHP; NR0B2) is an atypical orphan nuclear receptor and acts as a coregulator of various nuclear receptors. Herein, we examined a novel cross talk between SHP and a forkhead transcription factor HNF3 (hepatocyte nuclear factor 3/Foxa. Transient transfection assay demonstrated that SHP inhibited the transcriptional activity of all three isoforms of HNF3, HNF3alpha, beta, and gamma. In vivo and in vitro protein interaction studies showed that SHP physically interacted with HNF3. Adenovirus-mediated overexpression of SHP significantly decreased the mRNA levels of glucose-6-phosphase (G6Pase), cholesterol 7-alpha-hydroxylase (CYP7A1), and phosphoenolpyruvate carboxykinase (PEPCK) in HepG2 cells and rat primary hepatocytes. Moreover, the mRNA level of G6Pase was notably increased by down-regulation of SHP with small interfering RNA. Interestingly, HNF3 transactivity was still repressed by SHPDelta128-139 that fails to repress nuclear receptors. Mapping of interaction domain revealed that SHP interacted with forkhead DNA binding domain of HNF3alpha. Gel mobility shift and chromatin immunoprecipitation assays demonstrated that SHP inhibits DNA binding of HNF3. These results suggest that SHP is involved in the regulation of G6Pase, CYP7A1, and PEPCK gene expression via novel mechanism of inhibition of HNF3 activity and expand the role of SHP as a coregulator of other family of transcription factors in addition to nuclear receptors. PMID- 15358836 TI - Hormonal regulation of metastasis-associated protein 3 transcription in breast cancer cells. AB - Metastasis-associated protein 3 (MTA3) is a cell type-specific subunit of the Mi 2/NuRD transcriptional corepressor complex. In breast cancer cells, MTA3 and the Mi-2/NuRD complex mediate repression of Snail, a transcription factor that promotes epithelial to mesenchymal transitions. Thus, MTA3 functions to maintain a differentiated, epithelial status in breast cancer. Interestingly, in mammary epithelial cells, MTA3 biosynthesis requires both functional estrogen receptor (ER) and estradiol. Here we have investigated the molecular basis for estrogen and ER-dependent expression of MTA3 in breast cancer cells. Molecular dissection of the MTA3 promoter using transient transfection assays identified a composite element required for high-level transcription consisting of an SP1 site in close proximity to a consensus estrogen response element half-site. Depletion of either SP1 or ER-alpha by RNA interference led to loss of MTA3 transcript in multiple breast cancer cell lines, indicating a requirement for both transcription factors in expression of endogenous MTA3. The MTA3 gene thus joins a growing list of loci regulated by both SP1 and ER. PMID- 15358838 TI - ALAMA: 25 years young. PMID- 15358837 TI - Canonical Wnt signaling is critical to estrogen-mediated uterine growth. AB - Major biological effects of estrogen in the uterus are thought to be primarily mediated by nuclear estrogen receptors, ERalpha and ERbeta. We show here that estrogen in an ER-independent manner rapidly up-regulates the expression of Wnt4 and Wnt5a of the Wnt family and frizzled-2 of the Wnt receptor family in the mouse uterus. One of the mechanisms by which Wnts mediate canonical signaling involves stabilization of intracellular beta-catenin. We observed that estrogen treatment prompts nuclear localization of active beta-catenin in the uterine epithelium. We also found that adenovirus mediated in vivo delivery of SFRP-2, a Wnt antagonist, down-regulates estrogen-dependent beta-catenin activity without affecting some of the early effects (water imbibition and angiogenic markers) and inhibits uterine epithelial cell growth, suggesting that canonical Wnt signaling is critical to estrogen-induced uterine growth. Our present results provide evidence for a novel role of estrogen that targets early Wnt/beta-catenin signaling in an ER-independent manner to regulate the late uterine growth response that is ER dependent. PMID- 15358839 TI - Seeking help in relation to work--visiting the occupational physician or the general practitioner. AB - BACKGROUND: It has not yet been examined whether employees with health or work problems prefer visiting the Occupational Physician (OP) or the General Practitioner (GP). AIM: Examining whether health and work problems predict visiting the OP or GP. METHOD: Multiple regression analyses within a prospective cohort study. RESULTS: None of the predictors was exclusively associated with visiting the OP, while emotional work demands and work-family conflict were associated with visiting the GP in relation to work. CONCLUSIONS: OPs might wish to clarify their preventive role to employees. PMID- 15358840 TI - Active or passive treatment for neck-shoulder pain in occupational health care? A randomized controlled trial. AB - AIMS: To compare the effectiveness of thoracic manipulations with instructions for physiotherapeutic exercises for the treatment of neck pain in occupational health care. METHODS: Seventy-five subjects aged 30-55 years from a random sample of 241 employees of the Finnish Broadcasting Company were randomly allocated to treatment in the form of four thoracic manipulations (n = 43), or instructions for physiotherapeutic exercises (n = 32). The subjects reported neck-shoulder pain on a structured pain questionnaire using a visual analogue scale (VAS, 0 10). Muscle tenderness and tender thoracic levels were evaluated by a blinded investigator (A.S.) at 6- and 12 month follow-ups. RESULTS: A statistically significant difference was found in self-reported worst pain by VAS at the 12 month follow-up in favour of the thoracic manipulation group. CONCLUSIONS: The natural course of the neck-shoulder pain in this study appears benign; pain was also reduced in the drop-out group. Both treatments were found effective at the 12 month follow-up. The effect of four manipulations was more favourable than the personal exercise program in treating the more intense phase of pain. PMID- 15358841 TI - Treating physicians' perceptions of barriers to return to work of their patients in Southern Ontario. AB - AIMS: The aim of this paper is to understand the treating physician's perspective with respect to the barriers that their patients face returning to work from injury and illness. METHODS: The methodology used was focus groups conducted in Southern Ontario with treating physicians. RESULTS: The main barrier identified by the treating physicians in the return to work process was the lack of accommodated work. The main areas identified for the use of additional resources was facilitated investigations, assessments and treatments for their patients and education and training for treating physicians. With respect to communication, physicians indicated that they wanted to know more about the work to which their patients were returning. CONCLUSIONS: Treating physicians believe that the most significant barriers for the timely return to work for their patients exist in the workplace, specifically related to lack of knowledge about appropriate modified work. The treating physicians' role in the return to work process is demanding due to insufficient time to deal with return to work issues, lack of training, not enough of the appropriate information and the treating physicians' role ambiguity. PMID- 15358842 TI - Carotid artery dissection following minimal postural trauma in a firefighter. AB - Carotid artery dissections (CAD) are uncommon, but not rare, and are increasingly recognized as a cause of morbidity. A case of CAD following minimal sustained postural trauma is described. The causes and outcomes of CAD are discussed, with particular reference to risks that might be found in the workplace. PMID- 15358843 TI - Occupational asthma after exposure to plaster casts containing methylene diphenyl diisocyanate. AB - BACKGROUND: The case of a 34-year-old female nurse is presented. She worked in an accident and emergency department in a district general hospital, with methylene diphenyl diisocyanate (MDI)-containing synthetic plaster casts. She worked with MDI on a daily basis for 4 years. She was out of the department for 1 year and on her return developed cough, wheeze and dyspnoea within 5 min of exposure to MDI- containing synthetic casts. METHOD: A bronchial provocation test was performed and confirmed an early asthmatic response. RESULTS: There was a 39% decrease in the forced expiratory volume in 1 s 15 min after exposure, which required the administration of a bronchodilator on two occasions. The patient has subsequently avoided MDI-containing synthetic plaster casts and has experienced no further respiratory symptoms. CONCLUSION: This case illustrates that respiratory sensitization can occur as a result of exposure to MDI-containing synthetic casts and highlights the need for vigilance when health care workers are using isocyanate-containing synthetic casts. PMID- 15358844 TI - Thyroiditis and inflammatory bowel disease associated with 50 Hz magnetic field exposure. PMID- 15358845 TI - Monitor. Mesothelioma deaths in Great Britain. PMID- 15358846 TI - Computer-aided detection performance in mammographic examination of masses: assessment. AB - PURPOSE: To compare performance of two computer-aided detection (CAD) systems and an in-house scheme applied to five groups of sequentially acquired screening mammograms. MATERIALS AND METHODS: Two hundred nineteen film-based mammographic examinations, classified into five groups, were included in this study. Group 1 included 58 examinations in which verified malignant masses were detected during screening; group 2, 39 in which all available latest examinations were performed prior to diagnosis of these malignant masses (subset of 39 women from group 1); group 3, 22 in which findings were interpreted as negative but were verified as cancer within 1 year from the negative interpretation (missed cancers); group 4, 50 in which findings were negative and patients were not recalled for additional procedures; and group 5, 50 in which patients were recalled for additional procedures and findings were negative for cancer. In all examinations, images were processed with two Food and Drug Administration-approved commercially available CAD systems and an in-house scheme. Performance levels in terms of true positive detection rates and number of false-positive identifications per image and per examination were compared. RESULTS: Mass detection rates in positive examinations (group 1) were 67%-72%. Detection rates among three systems were not significantly different (P > .05). In 50 negative screening examinations (group 4), false-positive rates ranged from 1.08 to 1.68 per four-view examination. Performance level differences among systems were significant for false-positive rates (P = .008). Performance of all systems was at levels lower than publicly suggested in some retrospective studies. False-positive CAD cueing rates were significantly higher for negative examinations in which patients were recalled (group 5) than they were for those in which patients were not recalled (group 4) (P < or = .002). CONCLUSION: Performance of CAD systems for mass detection at mammography varies significantly, depending on examination and system used. Actual performance of all systems in clinical environment can be improved. PMID- 15358847 TI - Radiation dose and image quality in pediatric CT: effect of technical factors and phantom size and shape. AB - PURPOSE: To evaluate effects of varying tube current and voltage on radiation dose, image noise, and image contrast with different phantom sizes and shapes. MATERIALS AND METHODS: Four round lucite phantoms with 8-32-cm diameters were scanned with multi-detector row computed tomography (CT) and 80-120 kVp. Radiation dose was based on CT dose index, image noise, and iodine contrast and measured with constant and variable tube currents that were age appropriate for each tube voltage. Radiation dose and image noise and contrast were compared in round and oval 24-cm phantoms. For various combinations of technical factors and phantom sizes and shapes, percentage differences were calculated for radiation dose and image noise and contrast. Associations between tube voltage and radiation dose, image noise, and image contrast in round and oval phantoms were determined by fitting second-degree polynomials to data. Differences in radiation dose and image noise and contrast, which were attributable to differences in tube voltage, were tested with paired t tests. RESULTS: With 165-mAs tube current, radiation doses with 140- and 80-kVp tube voltages were 103% ([41.9 mGy - 20.6 mGy]/20.6 mGy) and 58% ([10.2 mGy - 4.2 mGy]/10.1 mGy) higher in the 8-cm phantom than in the 32-cm phantom. When tube current was adapted for phantom size, radiation dose at 80 kVp in the 8-cm phantom was reduced by 82% ([10.1 mGy - 1.8 mGy]/10.1 mGy). In the 8-cm phantom, tube voltage was decreased from 120 to 80 kVp and tube current remained at 165 mAs, resulting in a 68% noise increase ([3.1 HU - 1.8 HU]/1.8 HU). With variable tube current, 80-kVp tube voltage in the 8-cm phantom led to a 138% noise increase ([7.3 HU - 3.1 HU]/3.1 HU). With reduced tube voltage, image contrast increased. In the 8-cm phantom, with a constant 165 mAs tube current and a decrease in tube voltage from 120 to 80 kVp, there was a 35% ([333 HU - 217 HU]/333 HU) increase in contrast. No difference was noted in radiation dose or noise between round and oval phantoms (P = .604 and P = .06, respectively), but a small statistically significant difference (1%) in contrast attenuation was demonstrated (P = .025). CONCLUSION: Reduced tube voltage for pediatric contrast material-enhanced CT reduces radiation dose and maintains image contrast. Image noise increases, but the effect is minimal in smaller phantoms. An additional reduction in tube current further reduces radiation dose. PMID- 15358848 TI - MR coronary angiography with SH L 643 A: initial experience in patients with coronary artery disease. AB - PURPOSE: To prospectively assess the accuracy of breath-hold three-dimensional magnetic resonance (MR) coronary angiography with the gadolinium-based intravascular contrast agent SH L 643 A in patients with coronary artery disease. MATERIALS AND METHODS: Twelve patients (seven men, five women; age range, 46-78 years; mean age, 61.3 years) with angiographically proved coronary artery disease (luminal narrowing >50%) underwent breath-hold three-dimensional MR coronary angiography before and after injection of SH L 643 A (0.1 mmol gadolinium per kilogram body weight). For all MR examinations, signal-to-noise ratio and contrast-to-noise ratio were measured. Image quality was assessed with a four point scale. Conventional angiograms and MR angiograms were evaluated for depiction of the left main, proximal and middle left anterior descending, proximal left circumflex, and proximal and middle right coronary artery segments in a blinded fashion by two experienced readers in consensus. Results of this evaluation were compared by using a paired Student t test. P < .05 was considered to indicate a statistically significant difference. RESULTS: For the 72 coronary artery segments, the contrast-to-noise ratio significantly improved after administration of SH L 643 A, compared with the prior ratio (9.8 +/- 5.1 [standard deviation] vs 23.0 +/- 8.7; P < .01), whereas the difference in signal to-noise ratio did not reach statistical significance (25.2 +/- 11.4 vs 29.5 +/- 9.8; P > .3). Image quality significantly improved from a mean of 2.0 +/- 0.9 for nonenhanced images to 2.9 +/- 0.9 (P < .03) for contrast material-enhanced images. The proportion of segments for which images were nondiagnostic decreased from 38% to 10% with application of SH L 643 A. Overall sensitivity and specificity of contrast-enhanced MR coronary angiography for detection of coronary artery disease were 80% and 93%, respectively, and accuracy was 87%. CONCLUSION: Use of SH L 643 A improves detection of coronary artery disease at three-dimensional MR coronary angiography. PMID- 15358849 TI - Frozen shoulder: MR arthrographic findings. AB - PURPOSE: To evaluate the magnetic resonance (MR) arthrographic findings in patients with frozen shoulder. MATERIALS AND METHODS: Preoperative MR arthrograms of 22 patients (six women, 16 men; mean age, 54.7 years) with frozen shoulder treated with arthroscopic capsulotomy were compared with arthrograms of 22 age- and sex-matched control subjects without frozen shoulder. The thickness of the coracohumeral ligament (CHL) and the joint capsule, as well as the volume of the axillary recess, were measured (Mann-Whitney test). Abnormalities in the CHL, subcoracoid fat, superior glenohumeral ligament, superior border of the subscapularis tendon, long biceps tendon, and subscapularis recess were analyzed in consensus by two blinded radiologists (chi(2) test). RESULTS: Patients with frozen shoulder had a significantly thickened CHL (4.1 mm vs 2.7 mm in controls) and a thickened joint capsule in the rotator cuff interval (7.1 mm vs 4.5 mm; P < .001 for both comparisons, Mann-Whitney test) but not in the axillary recess. The volume of the axillary recess was significantly smaller in patients with frozen shoulder than in control subjects (P = .03, Mann-Whitney test). Thickening of the CHL to 4 mm or more had a specificity of 95% and a sensitivity of 59% for diagnosis of frozen shoulder. Thickening of the capsule in the rotator cuff interval to 7 mm or more had a specificity of 86% and a sensitivity of 64%. Synovitis-like abnormalities at the superior border of the subscapularis tendon were significantly more common in patients with frozen shoulder than in control subjects (P = .014, chi(2) test). Complete obliteration of the fat triangle between the CHL and the coracoid process (subcoracoid triangle sign) was specific (100%) but not sensitive (32%). CONCLUSION: Thickening of the CHL and the joint capsule in the rotator cuff interval, as well as the subcoracoid triangle sign, are characteristic MR arthrographic findings in frozen shoulder. PMID- 15358850 TI - Computer-aided detection in screening mammography: variability in cues. AB - PURPOSE: To evaluate the variability of true-positive and false-positive cues by using a commercially available computer-aided detection (CAD) system for analysis of 50 malignancies in a screening population. MATERIALS AND METHODS: Fifty breast cancers detected at screening were analyzed by using a commercially available CAD system. Mean patient age was 62.2 years. Each set of mammograms (craniocaudal and mediolateral oblique views) was digitized and analyzed by the CAD system 10 times. One radiologist compared CAD output with the location of the malignancy at mammography and determined whether each lesion was marked accurately in one mammographic view, both views, or neither. Sensitivity and reproducibility of the CAD system were determined for both case- and image-based analysis. RESULTS: Overall sensitivity of the CAD system when at least one of the two mammographic views was marked correctly (case-base sensitivity) was 82.4%. Sensitivity when each mammographic view was considered separately (image-based sensitivity) was 61.1%. For case-based analysis, variability in true-positive CAD cues was demonstrated for 14 of 50 (28%) cases. For image-based analysis, inconsistency in CAD output was observed in 33 of 100 (33%) mammographic views that contained malignancies detected at screening. However, the CAD system consistently detected 40-43 of the 50 breast cancers in each of the 10 CAD runs. Variability for false positive marks was significantly greater than that for true-positive marks. CONCLUSION: Inconsistency was demonstrated for CAD analysis of breast cancers detected at screening. However, the CAD system was reasonably consistent in the overall number of cancers identified from run to run. Greater variability of the CAD system was also demonstrated for false-positive marks, as compared with true positive marks. PMID- 15358851 TI - Increased time rate of change of gradient fields: effect on peripheral nerve stimulation at clinical MR imaging. AB - PURPOSE: To increase gradient stimulation from 100% to a fixed 120% level and to assess patient acceptance of the degree of peripheral nerve stimulation (PNS) at magnetic resonance (MR) imaging. MATERIALS AND METHODS: Two hundred ten patients underwent MR imaging of various body regions according to clinical indications. An additional three-dimensional fast low-angle shot sequence with the 120% stimulation level was performed. A patient questionnaire was distributed after MR imaging to document the presence, degree, and location of PNS. Degree was measured with an 11-point scale (score range, 0-10). Age was analyzed between the sexes for significant statistical differences. Furthermore, correlation between location of examination and location and degree of stimulation was performed. To determine stimulation discomfort relative to other factors typically present at MR imaging, the degree of discomfort due to room temperature, size of magnet bore, acoustic noise, examination time, and heating sensation was determined for comparison, as well. RESULTS: Thirty-five (16.7%) patients reported a stimulation sensation during imaging in one or more locations, while six (2.9%) felt very uncomfortable local stimulation (score of 8-10). No significant difference between male and female patients regarding age, sex, and appearance or degree of stimulation sensation could be detected. No significant correlation between location of examination and location and degree of stimulation was recorded. Compared with other side effects, PNS was considered relatively unimportant. CONCLUSION: The 120% gradient stimulation level seems acceptable for routine clinical imaging with this gradient system, since only 2.9% of patients experienced very uncomfortable local stimulation. PMID- 15358852 TI - Spinal cord feeding arteries at MR angiography for thoracoscopic spinal surgery: feasibility study and implications for surgical approach. AB - PURPOSE: To prospectively investigate the feasibility of contrast material enhanced magnetic resonance (MR) angiography for visualization of the spinal vasculature in patients referred for video-assisted thoracoscopic surgical treatment of a thoracic herniated disk and to prospectively assess the influence of preoperative imaging of the spinal vasculature on the choice of surgical approach. MATERIALS AND METHODS: Eight patients (three men and five women; mean age, 58 years; range, 42-83 years) with a thoracic herniated disk underwent contrast-enhanced MR angiography of the thoracoabdominal aorta and posterior branches. Imaging was performed with three-dimensional first-pass contrast enhanced MR angiographic technique and a triple dose of gadolinium-based contrast agent. Images were analyzed by two observers in consensus to localize the Adamkiewicz artery (AKA) and its connections to the aorta and the anterior spinal artery (ASA). This information was used to determine any change in surgical approach. RESULTS: In all eight patients, the AKA, the ASA, and the connections with the aorta were identified. The AKA originated between T9 and L2 in all patients and derived from the left side of the aorta in 75% (six of eight) of the patients. In three patients in whom the AKA was observed on the left side, the surgical approach was changed to the right side to preserve spinal cord integrity. CONCLUSION: Preoperative imaging of the AKA is feasible with contrast enhanced MR angiography. Contrast-enhanced MR angiography can be used to image the main feeding arteries of the spinal cord in patients undergoing thoracoscopic spinal surgery, and results can be used to change the side of surgical approach. PMID- 15358853 TI - Peripheral arterial disease: therapeutic confidence of CT versus digital subtraction angiography and effects on additional imaging recommendations. AB - PURPOSE: To compare multi-detector row computed tomographic (CT) angiography and digital subtraction angiography (DSA) prior to revascularization in patients with symptomatic peripheral arterial disease for the purpose of assessing recommendations for additional imaging and physician confidence ratings for chosen therapy. MATERIALS AND METHODS: In a randomized controlled trial, 73 patients were assigned to CT angiography, and 72 were assigned to DSA. Physician confidence in the treatment decision was measured as a continuous outcome on a scale of 0-10 (uncertain to certain) and as a dichotomous outcome (further imaging recommended, yes or no). Mean confidence scores and additional imaging recommendations were compared between CT and DSA groups in an intention-to diagnose-and-treat analysis. To detect trends in confidence, confidence scores were plotted over time, and multiple linear regression analysis was performed. To detect trends in additional imaging recommendations, logistic regression analysis was used. Data from eligible nonrandomized patients were analyzed separately. RESULTS: No statistically significant difference in baseline characteristics between randomized groups was found. CT had a lower confidence score than did DSA (7.2 vs 8.2, P < .001). Further imaging was recommended more often after CT (25 of 71 patients, 35%) than after DSA (nine of 66 patients, 14%; P = .003). Analysis of trends demonstrated increasing (but not statistically significant) confidence in CT and stable confidence in DSA. No significant difference was found in baseline characteristics between randomized and nonrandomized patients. Among nonrandomized patients, no significant difference in mean confidence score (8.2 vs 8.3, P = .26) was found between CT (n = 24) and DSA (n = 26). CONCLUSION: With CT angiography, physician confidence decreases with an associated increase in additional imaging prior to revascularization in patients with symptomatic peripheral arterial disease. Given that CT is less invasive than DSA, results suggest that CT may replace DSA in selected cases. PMID- 15358854 TI - CT colonography and colonoscopy: assessment of patient preference in a 5-week follow-up study. AB - PURPOSE: To prospectively evaluate short- and midterm patient preference of computed tomographic (CT) colonography relative to colonoscopy in patients at increased risk for colorectal cancer and to elucidate determinants of preference. MATERIALS AND METHODS: Consecutive patients at increased risk for colorectal cancer underwent CT colonography prior to scheduled colonoscopy. Patient experience and preference were assessed both directly after the examinations and 5 weeks after the examinations. Differences in pain, embarrassment, discomfort, and preference were assessed with the Wilcoxon signed rank sum test or a binomial test. Potential determinants of preference were investigated with logistic regression analyses. RESULTS: Data for 249 patients were included. Fewer patients experienced severe or extreme pain during CT colonography (seven [3%] of 245) than during colonoscopy (81 [34%] of 241) (P < .001). Directly after both examinations, 168 (71%) of 236 patients preferred CT colonography; 5 weeks later, 141 (61%) of 233 patients preferred CT colonography (P < .001). Initially, a painful colonoscopy examination (odds ratio, 0.17; 95% confidence interval [CI]: 0.08, 0.38) was a determinant of CT colonography preference. Similarly, a painful (odds ratio, 3.70; 95% CI: 1.54, 8.92) or an embarrassing (odds ratio, 4.46; 95% CI: 1.18, 16.88) CT colonography examination was a determinant of colonoscopy preference. After 5 weeks, the presence of polyps emerged as a determinant of colonoscopy preference (odds ratio, 1.94; 95% CI: 1.02, 3.70), while the role of experiences waned. CONCLUSION: Patients preferred CT colonography to colonoscopy; however, this preference decreased in time, while outcome considerations gradually replaced temporary experiences of inconvenience. PMID- 15358855 TI - Traumatic neuroma and recurrent lymphadenopathy after neck dissection: comparison of radiologic features. AB - PURPOSE: To retrospectively evaluate the ultrasonographic (US), computed tomographic (CT), and magnetic resonance (MR) imaging features that differentiate traumatic neuroma from recurrent lymphadenopathy after neck dissection. MATERIALS AND METHODS: Imaging findings of 10 patients with a traumatic neuroma and 17 with recurrent lymphadenopathy were reviewed. US and CT were performed in all patients; MR imaging was performed in 16 patients. Findings analyzed at US included the diameter of the long and short axes, the short-axis-to-long-axis ratio, and the presence of a central hyperechoic area. Findings analyzed at CT were contiguity with common or internal carotid artery, lesion location in correlation with carotid artery, and the presence of a hyperattenuating rim. Findings analyzed at MR imaging included signal intensity on T1- and T2-weighted images, the presence of ring enhancement, and the presence of a hypointense rim on T2-weighted images. RESULTS: Statistically significant differences were found between traumatic neuroma and recurrent lymphadenopathy in the short-axis-to-long axis ratio (mean, 0.47 vs 0.72; P < .001), the short-axis diameter (mean, 5.7 vs 12.2 mm; P < .001), the presence of a central hyperechoic area (five of 10 patients [50%] vs one of 17 patients [6%]; P < .05), the frequency of contact with carotid artery (two of 10 patients [20%] vs 13 of 17 patients [76%]; P < .01), and the presence of a hypointense rim on T2-weighted MR images (three of six patients [50%] vs zero of 10 patients [0%]; P < .05). Findings in other parameters were not statistically significant. CONCLUSION: Several imaging findings can differentiate traumatic neuroma from recurrent lymphadenopathy after neck dissection. PMID- 15358856 TI - Rational design and selection of bivalent peptide ligands of thrombin incorporating P4-P1 tetrapeptide sequences: from good substrates to potent inhibitors. AB - The tetrapeptide Phe-Asn-Pro-Arg is a structurally optimized sequence for binding to the active site of thrombin. By conjugating this tetrapeptide or some variants to a C-terminal fragment of hirudin, we were able to generate a series of new bivalent inhibitors of thrombin containing only genetically encodable natural amino acids. We found that synergistic binding to both the active site and an exosite of thrombin can be enhanced through substitutions of amino acid residues at the P3 and P3' sites of the active-site directed sequence, Phe(P4)-Xaa(P3) Pro(P2)-Arg(P1)-Pro(P1')-Gln(P2')-Yaa(P3'). Complementary to rational design, a phage library was constructed to explore further the residue requirements at the P4, P3 and P3' sites for bivalent and optimized two-site binding. Very significantly, panning of the phage library has led to thrombin-inhibitory peptides possessing strong anti-clotting activities in the low nanomolar range and yet interfering only partially the catalytic active site of thrombin. Modes of action of the newly discovered bivalent inhibitors are rationalized in light of the allosteric properties of thrombin, especially the interplay between the proteolytic action and regulatory binding occurring at thrombin surfaces remote from the catalytic active site. PMID- 15358857 TI - Morphology and the internal structure of words. AB - Morphology is the aspect of language concerned with the internal structure of words, and languages vary in the extent to which they rely on morphological structure. Consequently, it is not clear whether morphology is a basic element of a linguistic structure or whether it emerges from systematic regularities between the form and meaning of words. Here, we looked for evidence of morphological structure at a neural systems level by using a visual masked priming paradigm and functional MRI. Form and meaning relations were manipulated in a 2 x 2 design to identify reductions in blood oxygenation level-dependent signal related to shared form (e.g., corner-corn), shared meaning (e.g., idea-notion), and shared morphemes (e.g., boldly-bold, which overlapped in both form and meaning). Relative to unrelated pairs (e.g., ozone-hero), morphologically related items reduced blood oxygenation level-dependent signal in the posterior angular gyrus bilaterally, left occipitotemporal cortex, and left middle temporal gyrus. In the posterior angular gyrus, a neural priming effect was observed for all three priming conditions, possibly reflecting reduced attentional demands rather than overlapping linguistic representations per se. In contrast, the reductions seen in the left occipitotemporal cortex and left middle temporal gyrus corresponded, respectively, to main effects of orthographic and semantic overlap. As neural regions sensitive to morphological structure overlapped almost entirely with regions sensitive to orthographic and semantic relatedness, our results suggest that morphology emerges from the convergence of form and meaning. PMID- 15358858 TI - Insights into the evolution of Yersinia pestis through whole-genome comparison with Yersinia pseudotuberculosis. AB - Yersinia pestis, the causative agent of plague, is a highly uniform clone that diverged recently from the enteric pathogen Yersinia pseudotuberculosis. Despite their close genetic relationship, they differ radically in their pathogenicity and transmission. Here, we report the complete genomic sequence of Y. pseudotuberculosis IP32953 and its use for detailed genome comparisons with available Y. pestis sequences. Analyses of identified differences across a panel of Yersinia isolates from around the world reveal 32 Y. pestis chromosomal genes that, together with the two Y. pestis-specific plasmids, to our knowledge, represent the only new genetic material in Y. pestis acquired since the the divergence from Y. pseudotuberculosis. In contrast, 149 other pseudogenes (doubling the previous estimate) and 317 genes absent from Y. pestis were detected, indicating that as many as 13% of Y. pseudotuberculosis genes no longer function in Y. pestis. Extensive insertion sequence-mediated genome rearrangements and reductive evolution through massive gene loss, resulting in elimination and modification of preexisting gene expression pathways, appear to be more important than acquisition of genes in the evolution of Y. pestis. These results provide a sobering example of how a highly virulent epidemic clone can suddenly emerge from a less virulent, closely related progenitor. PMID- 15358859 TI - Amine-synthesizing enzyme N-substituted formamide deformylase: screening, purification, characterization, and gene cloning. AB - N-substituted formamide was produced through the hydration of an isonitrile by isonitrile hydratase in the isonitrile metabolism. The former compound was further degraded by a microorganism, strain F164, which was isolated from soil through an acclimatization culture. The N-substituted formamide-degrading microorganism was identified as Arthrobacter pascens. The microbial degradation was found to proceed through an enzymatic reaction, the N-substituted formamide being hydrolyzed to yield the corresponding amine and formate. The enzyme, designated as N-substituted formamide deformylase (NfdA), was purified and characterized. The native enzyme had a molecular mass of approximately 61 kDa and consisted of two identical subunits. It stoichiometrically catalyzed the hydrolysis of N-benzylformamide (an N-substituted formamide) to benzylamine and formate. Of all of the N-substituted formamides tested, N-benzylformamide was the most suitable substrate for the enzyme. However, no amides were accepted as substrates. The gene (nfdA) encoding this enzyme was also cloned. The deduced amino acid sequence of nfdA exhibited the highest overall sequence identity (28%) with those of regulatory proteins among known proteins. Only the N-terminal region (residues 58-72) of NfdA also showed significant sequence identity (27 73%) to that of each member of the amidohydrolase superfamily, although there was no similarity in the overall sequence except in the above limited region. PMID- 15358860 TI - Irregularity, volatility, risk, and financial market time series. AB - The need to assess subtle, potentially exploitable changes in serial structure is paramount in the analysis of financial data. Herein, we demonstrate the utility of approximate entropy (ApEn), a model-independent measure of sequential irregularity, toward this goal, by several distinct applications. We consider both empirical data and models, including composite indices (Standard and Poor's 500 and Hang Seng), individual stock prices, the random-walk hypothesis, and the Black-Scholes and fractional Brownian motion models. Notably, ApEn appears to be a potentially useful marker of system stability, with rapid increases possibly foreshadowing significant changes in a financial variable. PMID- 15358861 TI - A complex between peptide:N-glycanase and two proteasome-linked proteins suggests a mechanism for the degradation of misfolded glycoproteins. AB - Peptide:N-glycanase (PNGase) has been proposed to participate in the proteasome dependent glycoprotein degradation pathway. The finding that yeast PNGase interacts with the 19S proteasome subunit through the protein Rad23 supports this hypothesis. In this report, we have used immunofluorescence, subcellular fractionation, coimmunoprecipitation, and in vitro GST pull-down techniques for detecting intracellular localization and interactions of PNGase, HR23B, and S4 by using human (h) and mouse (m) homologs. Immunofluorescence studies revealed that hPNGase, hHR23B, and hS4 are present in close proximity to the endoplasmic reticulum (ER) when calnexin was used as an ER marker in HeLa cells. Subcellular fractionation suggests not only cytoplasmic but also ER association of hPNGase in HeLa cells. Immunoprecipitation analysis revealed the interaction of h/mPNGase with the 19S proteasome subunit, hS4, through hHR23B. Using an in vitro GST pull down assay, we also have shown that recombinant mPNGase requires its N terminus and middle domain for interaction with mHR23B. Finally, using misfolded yeast carboxypeptidase Y and chicken ovalbumin as glycoprotein substrates, we have established that mHR23B acts as a receptor for deglycosylated proteins. Based on this finding, we propose that after deglycosylation of misfolded glycoproteins by PNGase, the aglyco forms of these proteins are recognized by HR23B and targeted for degradation. PMID- 15358862 TI - Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension. AB - Pulmonary vascular medial hypertrophy caused by excessive pulmonary artery smooth muscle cell (PASMC) proliferation is a major cause for the elevated pulmonary vascular resistance in patients with idiopathic pulmonary arterial hypertension (IPAH). Increased Ca(2+) influx is an important stimulus for PASMC proliferation. Transient receptor potential (TRP) channel genes encode Ca(2+) channels that are responsible for Ca(2+) entry during cell proliferation. Normal human PASMC expressed multiple canonical TRP (TRPC) isoforms; TRPC6 was highly expressed and TRPC3 was minimally expressed. The protein expression of TRPC6 in normal PASMC closely correlated with the expression of Ki67, suggesting that TRPC6 expression is involved in the transition of PASMC from quiescent phase to mitosis. In lung tissues and PASMC from IPAH patients, the mRNA and protein expression of TRPC3 and -6 were much higher than in those from normotensive or secondary pulmonary hypertension patients. Inhibition of TRPC6 expression with TRPC6 small interfering RNA markedly attenuated IPAH-PASMC proliferation. These results demonstrate that expression of TRPC channels correlates with the progression of the cell cycle in PASMC. TRPC channel overexpression may be partially responsible for the increased PASMC proliferation and pulmonary vascular medial hypertrophy in IPAH patients. PMID- 15358863 TI - A balance between excitatory and inhibitory synapses is controlled by PSD-95 and neuroligin. AB - Factors that control differentiation of presynaptic and postsynaptic elements into excitatory or inhibitory synapses are poorly defined. Here we show that the postsynaptic density (PSD) proteins PSD-95 and neuroligin-1 (NLG) are critical for dictating the ratio of excitatory-to-inhibitory synaptic contacts. Exogenous NLG increased both excitatory and inhibitory presynaptic contacts and the frequency of miniature excitatory and inhibitory synaptic currents. In contrast, PSD-95 overexpression enhanced excitatory synapse size and miniature frequency, but reduced the number of inhibitory synaptic contacts. Introduction of PSD-95 with NLG augmented synaptic clustering of NLG and abolished NLG effects on inhibitory synapses. Interfering with endogenous PSD-95 expression alone was sufficient to reduce the ratio of excitatory-to-inhibitory synapses. These findings elucidate a mechanism by which the amounts of specific elements critical for synapse formation control the ratio of excitatory-to-inhibitory synaptic input. PMID- 15358865 TI - Jun turnover is controlled through JNK-dependent phosphorylation of the E3 ligase Itch. AB - The turnover of Jun proteins, like that of other transcription factors, is regulated through ubiquitin-dependent proteolysis. Usually, such processes are regulated by extracellular stimuli through phosphorylation of the target protein, which allows recognition by F box-containing E3 ubiquitin ligases. In the case of c-Jun and JunB, we found that extracellular stimuli also modulate protein turnover by regulating the activity of an E3 ligase by means of its phosphorylation. Activation of the Jun amino-terminal kinase (JNK) mitogen activated protein kinase cascade after T cell stimulation accelerated degradation of c-Jun and JunB through phosphorylation-dependent activation of the E3 ligase Itch. This pathway modulates cytokine production by effector T cells. PMID- 15358864 TI - Dynamic antagonism between RNA-binding protein CUGBP2 and cyclooxygenase-2 mediated prostaglandin E2 in radiation damage. AB - Damage to intestinal epithelium limits the use of ionizing radiation (IR) in cancer therapy. Prostaglandins (PGs), generated through the action of cyclooxygenase-1 (COX-1) and COX-2 protect the intestinal stem cells from IR. In previous studies, we demonstrated that the RNA-binding protein CUGBP2 regulates the stability and translation of COX-2 mRNA by interacting with AU-rich sequences in 3' UTR. Here, we demonstrate a dynamic antagonistic relationship between CUGBP2 and COX-2. Both CUGBP2 and COX-2 are rapidly induced after IR in intestinal crypt epithelial cells in mice, but CUGBP2 protein expression is observed immediately and COX-2 protein expression is delayed. In contrast, administration of bacterial lipopolysaccharide induced COX-2 expression and PGE(2), resulting in the inhibition of CUGBP2 expression and radioprotection of the intestine. These effects were reversed by NS398, a COX-2-specific inhibitor, suggesting that lipopolysaccharide-mediated inhibition of CUGBP2 is a PG dependent mechanism. Furthermore, CUGBP2 expression is higher in COX-1(-/-) and COX-2(-/-) mice than wild-type controls at basal conditions, which is further increased after IR. PMID- 15358866 TI - Single-atom spin-flip spectroscopy. AB - We demonstrate the ability to measure the energy required to flip the spin of single adsorbed atoms. A low-temperature, high-magnetic field scanning tunneling microscope was used to measure the spin excitation spectra of individual manganese atoms adsorbed on Al2O3 islands on a NiAl surface. We find pronounced variations of the spin-flip spectra for manganese atoms in different local environments. PMID- 15358867 TI - Controlling the dynamics of a single atom in lateral atom manipulation. AB - We studied the dynamics of a single cobalt (Co) atom during lateral manipulation on a copper (111) surface in a low-temperature scanning tunneling microscope. The Co binding site locations were revealed in a detailed image that resulted from lateral Co atom motion within the trapping potential of the scanning tip. Random telegraph noise, corresponding to the Co atom switching between hexagonal close packed (hcp) and face-centered cubic (fcc) sites, was seen when the tip was used to try to position the Co atom over the higher energy hcp site. Varying the probe tip height modified the normal copper (111) potential landscape and allowed the residence time of the Co atom in these sites to be varied. At low tunneling voltages (less than approximately 5 millielectron volts), the transfer rate between sites was independent of tunneling voltage, current, and temperature. At higher voltages, the transfer rate exhibited a strong dependence on tunneling voltage, indicative of vibrational heating by inelastic electron scattering. PMID- 15358868 TI - Superior visual search in adults with autism. AB - Recent studies have suggested that children with autism perform better than matched controls on visual search tasks and that this stems from a superior visual discrimination ability. This study assessed whether these findings generalize from children to adults with autism. Experiments 1 and 2 showed that, like children, adults with autism were superior to controls at searching for targets. Experiment 3 showed that increases in target-distractor similarity slowed the visual search performance of the control group significantly more than that of the autism group, suggesting that the adults with autism have a superior visual discrimination ability. Thus, these experiments replicate in adults previous findings in children with autism. Superior unique item detection in adults with autism, stemming from enhanced discrimination, is discussed in the light of the possible role of stimulus processing disturbances in the disorder in general. PMID- 15358869 TI - Advanced mind-reading in adults with Asperger syndrome. AB - This study investigated the mind-reading abilities of 19 adults with Asperger syndrome and 19 typically developing adults. Two static mind-reading tests and a more naturalistic empathic accuracy task were used. In the empathic accuracy task, participants attempted to infer the thoughts and feelings of target persons, while viewing a videotape of the target persons in a naturally occurring conversation with another person. The results are consistent with earlier findings. The empathic accuracy task indicated significant between-group differences, whereas no such differences were found on the static mind-reading tasks. The most innovative finding of the present study is that the inference ability of adults with pervasive developmental disorder (PDD) and controls depends on the focus of the target's thoughts and feelings, and that the empathic accuracy of adults with Asperger syndrome and control adults might be different in terms of quantity and quality. PMID- 15358870 TI - Weak central coherence: a cross-domain phenomenon specific to autism? AB - This study investigated whether evidence for the weak central coherence theory could be specifically associated with a group of children with autism compared with normally developing children (n=17 per group). Two tasks were employed, one involving visual illusions and the other verbal homophones. Both were based on tasks used in previous central coherence research. Incorporation of tasks involving the use of different domains (verbal versus visual) also enabled the investigation of claims that weak central coherence is a cross-domain processing style or deficit. The autistic group were found to be no different to the control group in performance on the visual illusions task. The autistic group made more errors than the normally developing group on the rare condition of the homophone task. However, analysis suggests this difference is mediated by verbal ability level and not diagnostic status per se. Theoretical implications and alternative explanations are discussed. PMID- 15358871 TI - Performance of children with autism spectrum disorder on advanced theory of mind tasks. AB - Although a number of advanced theory of mind tasks have been developed, there is sparse information on whether performance on different tasks is associated. The study examined the performance of 20 high-functioning 6- to 12-year-old children with autism spectrum disorder and 20 controls on three high-level theory of mind tasks: Strange Stories, Cartoons and the children's version of the Eyes task. The pattern of findings suggests that the three tasks may share differing, non specific, information-processing requirements in addition to tapping any putative mentalizing ability. They may also indicate a degree of dissociation between social-cognitive and social-perceptual or affective components of the mentalizing system. PMID- 15358872 TI - Parent management training and Asperger syndrome: a randomized controlled trial to evaluate a parent based intervention. AB - This controlled trial of a parent management intervention aimed to increase parental competence in management of problem behaviours associated with Asperger syndrome. The intervention compared two formats, a 1 day workshop and six individual sessions. Measures were taken on three occasions: pre-intervention, at 4 weeks, and at 3 month follow-up. Variables of interest were number and intensity of problem behaviours, and parent evaluation of social interaction skills. Results showed parents reporting fewer and lower intensity of problem behaviours and increased social interactions at 4 weeks and 3 months. Results held across formats and suggest that parent management training can provide an effective intervention for parents of a child with Asperger syndrome. Group differences on outcome measures and in the use of strategies are discussed along with limitations of the study. PMID- 15358873 TI - Comparison of behavioural and natural play interventions for young children with autism. AB - The article reports the results of a pilot study comparing traditional behavioural approaches and natural play interventions for young children with autism over a 10 week period. Two matched groups of eight young children with autism participated. Using a crossover design, children in both groups showed positive gains in compliance, attending, play and communication with their therapists and parents. Improvements in attending and compliance were higher following the behavioural condition compared with the natural play condition. Seven participants had reduced autism scores after the intervention. The findings suggest that behavioural and play approaches affect behaviour in different ways and that autistic symptomatology of young children may be amenable to treatment. The discussion focuses on the active ingredients of treatments and the need to base efficacy research on well-planned treatment comparisons. PMID- 15358874 TI - An attempt to define the core dysfunction of autism. PMID- 15358875 TI - Assessment of mindfulness by self-report: the Kentucky inventory of mindfulness skills. AB - A self-report inventory for the assessment of mindfulness skills was developed, and its psychometric characteristics and relationships with other constructs were examined. Participants included three samples of undergraduate students and a sample of outpatients with borderline personality disorder Based on discussions of mindfulness in the current literature, four mindfulness skills were specified: observing, describing, acting with awareness, and accepting without judgment. Scales designed to measure each skill were developed and evaluated. Results showed good internal consistency and test-retest reliability and a clear factor structure. Most expected relationships with other constructs were significant. Findings suggest that mindfulness skills are differentially related to aspects of personality and mental health, including neuroticism, psychological symptoms, emotional intelligence, alexithymia, experiential avoidance, dissociation, and absorption. PMID- 15358876 TI - The Five-factor Personality Inventory as a measure of the Five-factor Model: Belgian, American, and Hungarian comparisons with the NEO-PI-R. AB - The lexically based Five-Factor Personality Inventory (FFPI) was correlated with the factors and facets of the Revised NEO Personality Inventory (NEO-PI-R) in Belgian (N = 265), American (N = 116), and Hungarian (N = 320) samples. Results were similar across the three cultures. Analysis of orthogonalized FFPI factors showed that three of them--emotional stability, extraversion, and agreeableness- showed a direct correspondence to their NEO-PI-R counterparts. Autonomy, however, was not clearly related to openness, and facet analysis suggested that it might be interpreted as a dominance factor Better matches to NEO-PI-R conscientiousness and openness could be obtained by using vectors rotated 30 degrees from the FFPI positions. Raw scale scores showed similar results. Researchers should not assume that all measures of the Five-Factor Model are qualitatively similar PMID- 15358877 TI - Assessing stress in cancer patients: a second-order factor analysis model for the Perceived Stress Scale. AB - Using the Perceived Stress Scale (PSS), perceptions of global stress were assessed in 111 women following breast cancer surgery and at 12 and 24 months later This is the first study to factor analyze the PSS. The PSS data were factor analyzed each time using exploratory factor analysis with oblique direct quartimin rotation. Goodness-of-fit indices (root mean square error of approximation [RMSEA]), magnitude and pattern of factor loadings, and confidence interval data revealed a two-factor solution of positive versus negative stress items. The findings, replicated across time, also indicate factor stability. Hierarchical factor analyses supported a second-order factor of "perceived stress." This alternative factor model of the PSS is presented along with observations regarding the measure's use in cancer research. PMID- 15358878 TI - Evaluating the convergence of muscle appearance attitude measures. AB - There has been growing interest in the assessment of a muscular appearance. Given the importance of assessing muscle appearance attitudes, the aim of this study was to explore the convergence of the Drive for Muscularity Scale, Somatomorphic Matrix, Contour Drawing Rating Scale, Male Figure Drawings, and the Muscularity Rating Scale. Participants were 76 men and 103 women who each completed four measures of body image that were meant to evaluate two distinct domains of appearance attitudes: muscularity and body fat. For both genders, data indicated low convergence among muscularity measures, but high convergence among body fat measures. For men, the Drive for Muscularity Body Image subscale exhibited the greatest concurrent validity with behaviors used to increase muscularity. The results are discussed in light of the selection of methods for the assessment of muscle appearance attitudes. PMID- 15358879 TI - Development and testing of a new instrument for measuring concerns about dying in health care providers. AB - A new measure of concerns about dying was investigated in this psychometric study. The Concerns About Dying instrument (CAD) was administered to medical students, nursing students, hospice nurses, and life sciences graduate students (N = 207) on two occasions; on one occasion they also completed three related measures. Analyses included descriptive statistics, factor analysis, Cronbach's alpha, test-retest correlations, t tests, and correlations with other measures. Results suggest the CAD measures three distinct but related areas: general concern about death, spirituality, and patient-related concern about death. Reliability estimates were good, and correlations with related measures were strong. Between-group differences suggest scores are related to actual differences in level of concern and beliefs about death and dying. The CAD has the advantage of being very brief and of explicitly assessing concerns about working with patients who are dying. PMID- 15358880 TI - A comparison of four measures of self-control skills. AB - This study compares the psychometric characteristics of four questionnaires designed to assess self-control skills: the Self-Control Questionnaire, the Frequency of Self-Reinforcement Questionnaire, the Cognitive Self-Management Test, and the Lifestyle Approaches Inventory. Content validity was judged to be fairly comparable by three raters in Study 1. In Study 2, convergent and divergent validity support was obtained for all four questionnaires when administered to 369 multiethnic college students, but the relative degree of support varied across constructs. Hence, selection of a self-control instrument may be guided by the target behavior of interest. PMID- 15358881 TI - Construct validity of the MMPI-2 College Maladjustment (Mt) Scale. AB - The construct validity of the MMPI-2 (Minnesota Multiphasic Personality Inventory 2) College Maladjustment (Mt) Scale was examined using 376 student clients at a university psychological clinic. A principal components analysis and correlations of Mt scale scores with clients' and therapists' ratings of symptoms and functioning showed that the Mt scale identifies the presence of maladjustment as defined in terms of depressive and anxious symptoms. There is no evidence to show that the scale is specific to college students or that it is sensitive to severe psychological disturbance. The Mt scale does not inform the clinician as to why a person is distressed. In addition, there is no evidence from this study to suggest the superiority of the Mt scale over other MMPI-2 maladjustment measures. Therapists should use the entire MMPI-2 profile, not just the Mt scale, to gain the most comprehensive and specific understanding of clients. PMID- 15358882 TI - Three versions of the Interpersonal Adjective Scales and their fit to the circumplex model. AB - The Interpersonal Adjective Scales (IAS) is a well-supported instrument that is designed to map interpersonal traits onto the interpersonal circumplex. However, three versions of the IAS exist and these vary with respect to the degree to which they included item definitions (i.e., glossary added at the end, definitions attached to each item, and no definitions). The focus of this study was on comparing the structural similarity of each version. Each of the versions was given to different samples (N = 920) of college students. Results of multidimensional scaling (MDS) and randomized tests of order relations demonstrated that all versions were well and equally described by the circular model. There were no structural differences in the different IAS versions and thus each can be used with this population. PMID- 15358883 TI - If I have to say it one more time, I swear I'm gonna kill someone. PMID- 15358884 TI - Exploring nurses' experiences of providing high dependency care in children's wards. AB - The care of critically ill children has received much attention in the past decade, and many of the recommendations from reports relating to the provision of paediatric intensive care have been addressed. In comparison, high dependency care has received less attention. This level of care is often provided on 'general' children's wards, yet its impact on nursing staff and care delivery have received little attention. This article presents a study which explores nurses' experiences of providing high dependency care on children's wards using focus groups. Following analysis, six categories emerged: definitions and perceptions of high dependency care; having a high dependency unit; problems in district general hospitals; the skills needed for high dependency care; team working; and staffing for high dependency care. These findings have implications for current clinical, managerial and educational practice and for future developments in the field of high dependency care for children. PMID- 15358885 TI - Manual handling the deceased child in a children's hospice. AB - The introduction of the Manual Handling Operation Regulations 1992 has forced organizations to review all manual handling carried out in the work-place. Safe manual handling is now a legislative requirement to enable the workforce to carry out their jobs in a safe and structured environment. This project looked in detail at the manual handling that was being carried out when caring for a child after they have died. It was undertaken at CHASE Children's Hospice in the period up until December 2002, with a telephone survey investigating practice at six other children's hospices. It aimed to identify incorrect procedures and, by applying the theory of ergonomics, identify changes that needed to occur in practice, making recommendations for environmental changes. The study led to the production of a policy for handling the children, devising child-specific handling guidelines, reducing the number of times the children are moved after they have died and providing appropriate equipment. PMID- 15358886 TI - 'I had this horrible pain': the sources and causes of pain experiences in 4- to 11-year-old hospitalized children. AB - Researchers have relied, almost without exception, on adults for qualitative information about children's pain. However, adults may provide only a limited view of children's pain experience. The purpose of this article is to describe the events considered painful by children. Forty-four children participated in the study. They had been admitted for different reasons into different wards of a university hospital. The data consisted of qualitative child interviews and was analysed using inductive content analysis. The pain experience of children came from four main sources: 1. pain as a symptom of a diagnosed illness, 2. pain caused by medical and diagnostic procedures and basic nursing, 3. pain caused by accidents and 4. inexplicable pain not caused by a particular illness or injury. Children are able to report and describe their pain. Children should be regarded as experts on their pain in order to maximize the options for pain management. PMID- 15358887 TI - Evaluating student paediatric advanced nurse practitioners' experiences of a paediatric pharmacology module: in the context of reflective learning. AB - An evaluation of the first paediatric pharmacology module at Master's level was undertaken. The broad aims were: to ascertain whether the participants had achieved the modular learning outcomes; how their pharmacological knowledge base had altered as a result of the module; and whether their professional practice had been influenced subsequent to undertaking the module. The findings of the 12 participants who registered on the module are reported. Three tools were utilized to generate the data for this broadly qualitative study. The findings indicated that reflective thinking at the evaluative level enabled the participants to monitor their professional and personal performance and that their critical thinking skills and confidence had increased. The questionnaire survey had some limitations, but was seen as the only feasible method of achieving a representation as to whether the module had subsequently influenced the professional practice of the participants. The data presented convincing evidence of the relevance of study of a paediatric pharmacology module at Master's level for the professional and personal development of practitioners. PMID- 15358889 TI - Conceptualizing youth empowerment within tobacco control. AB - This article presents a conceptual framework that was developed to guide a national evaluation of the American Legacy Foundation's (Legacy) Statewide Youth Movement Against Tobacco Use (SYMATU) program. This program was designed to develop youth-led, youth-directed initiatives within local communities. Two evaluation studies were designed and implemented from 2000 through 2003: a cross site study that collected standard data elements across all 17 programs and a case study of five programs that collected formative data on variables thought to affect program implementation. In developing the youth empowerment (YE) conceptual framework, the authors started by reviewing literature to identify the concepts necessary for these types of initiatives and present a summary of their findings here. This article focuses on the development of the authors'overarching conceptual framework used to guide their evaluation studies. Other articles contained within this special issue present results from each of the SYMATU evaluation studies. PMID- 15358890 TI - Adult and group influences on participation in youth empowerment programs. AB - This study examined adult and group influences on youth participation in youth empowerment (YE) Statewide Youth Movement Against Tobacco Use programs. Adults provide individual guidance to youth groups and through existing tobacco control networks affect the social context in which YE programs are implemented. The authors tested hypotheses about the direct and indirect relationships between adults, groups, and youth collective participation. They used data from adults and youths to develop a measurement model of four higher order factors: adult involvement, group structure, group climate, and collective participation. They tested relationships between these factors in a structural equation model. Results showed that adult involvement did not have a meaningful direct effect on group factors or collective participation. Group factors mediated the relationship between adult involvement and collective participation. Further research is needed on how to measure adult involvement and its relationship to youth participation. PMID- 15358891 TI - Empowering organizations: approaches to tobacco control through youth empowerment programs. AB - Whereas most evaluations of youth empowerment focus on individual outcomes (i.e., were individual youths empowered?), this article focuses on the program as the unit of analysis and seeks to explain how organizational structures, program design features, and processes lead to organizational empowerment (OE). OE is defined as organizational efforts that generate psychological empowerment among members and organizational effectiveness needed for goal achievement. Case studies of five American Legacy Foundation-funded tobacco control youth empowerment programs were conducted during the first 2 years of implementation. Using an OE framework, the authors assessed program design features of the youth empowerment programs that contributed to or detracted from processes leading to OE. Comparing and contrasting the programs led to the identification of models and strategies that contribute to OE. Ecological influences of the state contexts (i.e., political climate, history of tobacco control, and public health infrastructure) were also examined. PMID- 15358892 TI - The North Carolina Youth Empowerment Study (NCYES): a participatory research study examining the impact of youth empowerment for tobacco use prevention. AB - This article describes the North Carolina Youth Empowerment Study (NC YES), a 3 year participatory evaluation of youth programs addressing tobacco use prevention. The study goals of NC YES were to (1) convene an advisory board comprised of lay youths and adults in a participatory research process, (2) document the characteristics of youth programs for tobacco use prevention and control in North Carolina, and (3) track the role of youth involvement in initiating and implementing 100% tobacco-free policies in local school districts. The NC YES Statewide Advisory Board helped guide the evaluation process, reviewed study protocols and data collection instruments, and helped interpret preliminary findings. Both quantitative and qualitative methods(e.g., telephone interviews and written questionnaires) were used to gather data from youth and adult leaders to achieve these aims. Lessons learned about the process of conducting participatory evaluation approaches and summary findings about the role of youths in policy advocacy efforts are presented. PMID- 15358893 TI - Quantifying the impact of participation in local tobacco control groups on the psychological empowerment of involved youth. AB - A core component of Legacy's Statewide Youth Movement Against Tobacco Use is the ability of state and local initiatives to empower youth to effect change in their communities. The authors' conceptual framework proposes that youth empowerment is an outcome of the process by which youths become active participants in local efforts. Youths are proposed to attain specific skills (e.g., assertiveness, advocacy), attitudes (e.g., domain-specific self-efficacy, perceived sociopolitical control, participatory competence), and knowledge of relevant resources. All are proposed outcomes of their individual participation in these local efforts. Data collected in fall 2002 through a tested survey instrument designed to obtain data on key components of empowerment are presented. Regression modeling was used to examine the extent to which characteristics of empowerment are an outcome of individual participation in these groups. A summary of lessons learned pertaining to effectively measuring empowerment and enhancing the empowerment process through local initiatives is provided. PMID- 15358894 TI - Examination of the relationship between community support and tobacco control activities as a part of youth empowerment programs. AB - Through the American Legacy Foundation's Statewide Youth Movement Against Tobacco Use (SYMATU), programs aimed at empowering youths to take action against tobacco use were funded. It is believed that the activities these groups undertake result in changes at the community level. This article examines the relationships between community support of tobacco control and the number and types of tobacco control-related activities these local youth groups conduct. Regression analyses examine the influence that different levels and sources of community support have on the quantity and focus of a group's activities. The influences of community support are briefly explored to understand if certain group characteristics have an impact on the quantity or sources of support for tobacco control. A deeper understanding of the importance and impact of community support will help groups understand the need for building community support networks and how these networks can assist them with implementation of program activities. PMID- 15358895 TI - Future directions for youth empowerment: commentary on application of youth empowerment theory to tobacco control. PMID- 15358896 TI - Views of health in the lay sector: a compilation and review of how individuals think about health. AB - The way lay people think about health and wellness influences their health and wellness-related behaviors. This article integrates and synthesizes the research conducted to understand lay health worldviews. We identify 18 themes that capture the findings about lay health worldviews. The themes fall into four categories: definitions of health, explanations for health, external and/or uncontrollable factors impinging on health, and the place health occupies in people's lives. The results lead to the observation that lay health worldviews--or health styles--are not understood in depth, particularly in US populations. Variation across the themes underscores the need for further descriptive research designed to understand consumers' ways of thinking about health and how the many changes in the professional and folk sectors affect lay worldviews. This has implications with respect to understanding consumers' health care behaviors and developing more effective communication strategies. PMID- 15358897 TI - Children's asthma experience and the importance of place. AB - Asthma is increasing dramatically among poor, inner-city children. Fourteen children with, and twelve without, asthma were interviewed and observed to examine how children with asthma understand their illness and how the asthma experience shapes their sense of place. Quantitative data showed no difference in objective preferences for particular places, but qualitative analysis showed important differences. We identified eight place-related themes, and on seven there were clear differences between children with and without asthma--offering freedom, being aesthetically pleasing, being comfortable, nature of social contact, being isolating, being familiar and being safe. Children experienced asthma as an interruption in daily life that influences time spent outside, being active and being with friends. Children with asthma are more sensitive to places that are 'dirty' or contain asthma triggers, are less likely to explore new or people-free places and emphasize family interactions, while children without asthma emphasize friendships and are sensitive to neighborhood violence. PMID- 15358898 TI - Managing safety and risk: the experiences of people with Parkinson's disease who use alternative and complementary therapies. AB - In this article I focus on how individuals living with Parkinson's disease manage safety and risk in their participation in alternative and complementary health care. I take a qualitative approach in this research, using semi-structured interviews and grounded theory techniques as means of generating and analysing data. My analysis centres on how these informants construct certain therapies as risk free and therefore safe, and others as risky and thus, inherently unsafe. I discuss the knowledge bases these informants draw on in their evaluations and describe the social contexts in which these assessments take place. While there is a substantial literature on risk in general, as well as the risk society, there has been less interest in micro-level analysis of risk. This article therefore contributes to knowledge through its focus on management of safety and risk in individuals' health-seeking behaviour. PMID- 15358899 TI - Constructing agency in treatment decisions: negotiating responsibility in cancer. AB - People belonging to cancer patient support groups participated in focus groups concerning their experiences of orthodox and complementary medicine. Their accounts of treatment decisions for cancer were analysed through discourse analysis. Accounts of both complementary and orthodox medicine addressed an ideological dilemma concerning the positioning of individuals as active or passive. Active positions were congruent with the everyday value of autonomy and responsible individuality, but conflicted with the established expertise of the medical profession in cancer and entailed being accountable for one's health. Passive positions reversed this situation. Complementary medicine provided an opportunity for people with cancer to negotiate active positions in a limited domain of health care. The responsibility for health associated with taking active treatment decisions was problematic in accounts of both orthodox and complementary medicine. PMID- 15358900 TI - Moral argumentation in talk about health and old age. AB - This study examines the ways in which people aged 90 or over construct and negotiate meanings of health in research interviews. Detailed analyses of two interview cases illustrate the flow of these negotiations within interviews, and how the interviewees balance different moral arguments. First, the interviewees try to manage the face-threat posed by the questioning about health. Second, their health accounts move on the axis of advocating good health and activity and conceding impaired health and diminished activity. They apply various rhetorical devices to challenge the traditional discourse of old age as decline. On the other hand, aligning oneself with the category of 'old' makes ill-health and inactivity legitimate, but brings along other kinds of moral obligations. Finally, the results are discussed in relation to cultural discourses of old age and health, and their moral implications. PMID- 15358901 TI - Risk ritual and the management of control and anxiety in medical culture. AB - Medical culture in advanced technological societies is characterized by an abundance of warnings about health hazards, along with an equally abundant flow of prescriptive advice for protecting individuals against them. Knowledge of health hazards also defines living and working environments and has spawned a politics of regulatory control. These features of contemporary health consciousness and action, along with deepening insecurities in the larger body politic, precipitate a spiral of anxiety and control. The spiral disrupts the presumed security derived from medical knowledge and medically informed behaviors and threatens to erode the boundaries of sanctioned action for health protection. In the context of the volatile political economy of health, the spiral, with its excesses of anxiety and demands for unauthorized controls, elicits efforts to contain it. I explore the symbolic dimensions of risk, what I call 'risk ritual'- the prevailing form of managing this troublesome contradiction. PMID- 15358903 TI - Designing and managing public housing self-sufficiency programs: the Youngs Lake Commons Program. AB - This article evaluates an experimental public housing self-sufficiency program that encourages home ownership among low-income families. A quasi-experimental design, in combination with focus groups, records review, and key informant interviews, provides data to focus on four questions: (a) Do these programs simply accelerate move-outs for those who would have left without intervention? (b) Are program elements replicable given the importance of the local context in public housing move-outs? (c) How do housing authorities resolve tensions that arise between housing management and social service delivery? (d) What should housing authority response be to those who fail? PMID- 15358904 TI - Institutionalization of a community action program targeting licensed premises in Stockholm, Sweden. AB - The objective of this article is to evaluate the level of institutionalization of a community action program targeting licensed premises in Stockholm. Five key factors were identified for institutionalization: adoption, sustainability, key leader support, structural changes, and compliance. A scale was developed to assess the strength of each factor. The results indicate a high degree of institutionalization (score 13 on a scale from 5 to 15). The authors conclude that the program activities have been sustained at a high level. A written agreement ensures that the participating organizations take responsibility for the continuing work. PMID- 15358905 TI - Firearm advertising: product depiction in consumer gun magazines. AB - In contrast to tobacco, alcohol, and other consumer products associated with health risks, we know very little about how firearm manufacturers advertise their products. The authors examined advertisements for firearms in all 27 ad-accepting magazines listed in Bacon's Magazine Directory "guns and shooting" category. Sixty-three manufacturers spent an estimated $1,195,680 on firearm advertising during the month studied. Annual advertising costs ranged widely; manufacturers spent an estimated $28.16 in advertising per firearm produced. Firearms generally were presented as a part of a lifestyle. Self-protection was noted infrequently in the advertisements. By contrast, attributes of the gun, typically technological characteristics, were noted in almost every advertisement. PMID- 15358906 TI - Alternative methods for handling attrition: an illustration using data from the Fast Track evaluation. AB - Using data from the evaluation of the Fast Track intervention, this article illustrates three methods for handling attrition. Multiple imputation and ignorable maximum likelihood estimation produce estimates that are similar to those based on listwise-deleted data. A panel selection model that allows for selective dropout reveals that highly aggressive boys accumulate in the treatment group over time and produces a larger estimate of treatment effect. In contrast, this model produces a smaller treatment effect for girls. The article's conclusion discusses the strengths and weaknesses of the alternative approaches and outlines ways in which researchers might improve their handling of attrition. PMID- 15358907 TI - The R word in Indian country: Culturally appropriate commercial tobacco-use research strategies. AB - With the highest tobacco abuse prevalence among all minorities, American Indians incur high rates of tobacco-related cancers. As a people, we have the poorest survival rate from cancer of any racial or ethnic group, due to a lack of access to specialist care and to low socioeconomic status (http://info.his.gov). With such epidemic abuse of commercial tobacco, we continuously lose our elders, adults, and children to disease and premature death. Therefore, it is essential to investigate theories of prevention, addiction, and cessation specifically related to our ethnicity. The authors of this article discuss past research wrongs and highlight culturally competent research strategies to aid Native communities in tobacco abuse prevention and education. The authors hope to contribute to bridging the gap between culturally relevant research and culturally relevant health promotion. PMID- 15358908 TI - Evaluation from start to finish: Incorporating comprehensive assessment into a training program for public health professionals. AB - This article examines the role of a comprehensive evaluation process in the context of a program to enhance use of the Internet by public health professionals. The Public Health Information Link for Nevada incorporated training and Web site development into a program to raise levels of awareness and competence of the public health professional. Evaluation played a key and ongoing role in the planning and implementation phases and made a marked contribution to the overall effectiveness of the project. Evaluation contributed to the ability to make midprogram adjustments, thereby enhancing user satisfaction and confidence. Participant evaluations indicated significant improvement in self perceived competence in utilizing the Internet for job-related activities. The authors conclude that the effort to integrate evaluation in an ongoing manner is balanced by the value of continuous feedback. A Web site review checklist, which will be useful to public health professionals who are developing new Web sites, is appended. PMID- 15358910 TI - Policy and environmental change: new directions for public health. AB - Solving major, persistent public health problems requires new policies and more aggressive, sweeping interventions that affect large populations. We need well conceived health policies and effective interventions for environmental change, but are we likely to get them? To find out, the Directors of Health Promotion and Education and the U.S. Centers for Disease Control and Prevention initiated a study of state and local public health agencies in the United States from 1996 through 1999. Data were collected by peer- and non-peer-reviewed literature searches, key informant interviews, reviews of Internet sites, and a nationwide survey. Study conclusions found confusion about the legitimacy of advocacy, lack of priority and funding for interventions that take more time versus quick fixes, variable leadership, reluctance to take risks, and a political climate that often discourages government agencies to take on these interventions. There are successes, yet more can be done. PMID- 15358911 TI - ATECAR: An Asian American community-based participatory research model on tobacco and cancer control. AB - In the past few decades, community-based participatory research, which underscores the indispensable role of the community in all phases of the research process, has been recognized as a viable approach to working constructively with communities to achieve mutually beneficial goals. This article presents a history of the Asian Tobacco Education, Cancer Awareness and Research's pioneering efforts in conducting community-based participatory research among Asian Americans in the Delaware Valley region of Pennsylvania and New Jersey. Information about project background, target populations, and the rationale for the conduct of community-based participatory research in American communities is provided. It also delineates the manner in which the principles of community based participatory research were applied as guides for the development of partnership infrastructures, research programs, and the challenges and barriers that were encountered. Facilitating factors in partnership building, and implications of employing this model in this ethnically and racially diverse population, are further discussed. PMID- 15358912 TI - A Bridge Between Communities: Video-making using principles of community-based participatory research. AB - Health educators can play a critical role in bringing together the partners and resources to successfully make videos using principles of community-based participatory research (CBPR). This article is a "how-to" guide for making videos using community-based participatory research principles. The authors describe video-making and CBPR, then outline six steps on how to make a video using principles of CBPR: (a) engaging stakeholders, (b) soliciting funding and informed consent, (c) creation of shared ownership, (d) building cross-cultural collaborations, (e) writing the script together, and (f) pulling it all together: editing and music selection. Still photographs and key themes from the video A Bridge Between Communities are presented as a running case study to illustrate these steps. The article concludes with implications for health promotion research and practice. PMID- 15358913 TI - Theoretical domains: a heuristic for teaching behavioral theory in HIV/STD prevention courses. AB - The HIV/STD epidemics have broadened the need for better behavioral intervention programs and highlighted the importance of providing training in behavioral theory to frontline program practitioners. However, there is a lack of effective methods for teaching theoretical concepts to people who may not have a background in behavioral science. This article presents a solution to this challenge by introducing a new heuristic for teaching theory and for placing individual theories/models in a broader context. Using a broad framework, we identify five domains that influence behavior: risk appraisal, self-perceptions, emotions and arousal, relationships and social influence, and environmental and structural factors. Each domain is described, and a brief overview of supporting literature is provided. Following the presentation of domains, we discuss course structure and activities. PMID- 15358914 TI - CHWs get credit: a 10-year history of the first college-credit certificate for community health workers in the United States. AB - Community health workers have become increasingly important in the U.S. health care system, playing a significant role in basic health promotion and care coordination; however, their status and visibility have not kept pace with their wider use. A major impediment has been the absence of systematic preparation-the field needs standardized education in programs that emphasize the actual skills and knowledge used by community health workers, programs that attract and retain nontraditional students from underserved communities and that foster professional advancement. This article chronicles the 10-year history of the first college credit-bearing community health worker certificate program in the country to address this need. Systematic research resulted in a program centered on the core competencies universally practiced by community health workers regardless of their topical focus. The certificate program combines performance-based methods with popular education into an innovative pedagogical approach that teaches skills, while solidifying, contextualizing, and enhancing crucial experiential knowledge. Program outcomes validate the approach. PMID- 15358915 TI - Recommendations from lead poisoning prevention program participants: best practices. AB - We present a program evaluation of the Phillips Lead Project, a 5-year study of the effectiveness of culture-specific, peer education in maintaining low blood lead levels of children in an inner-city neighborhood. We conducted focus groups to understand how project participants felt about the various strategies employed by the Lead Project. The purpose of this article is to describe their reactions to the project and make recommendations concerning appropriate educational strategies for lead poisoning prevention projects being undertaken in similar communities. Although this project was a research study, many of its methods, and participants' reactions to those methods, are relevant to non-research prevention programs and may be generalizable to other health issues besides lead poisoning. PMID- 15358916 TI - Viability of parks and recreation centers as sites for youth physical activity promotion. AB - This descriptive study explored the potential for public parks and recreation centers as intervention sites for promoting physical activity among youth. Directors (55% women) of 44 recreation centers in San Diego County completed a survey of their centers' physical activity programming for youths ages 3 to 17 years (response rate = 60%). On an average weekday, 373 (SD = 782) youths participated in physical activity at a center. More boys attended than girls (p <.05). Some centers sponsored after-school programs at elementary (41%) and middle (11%) schools but not at high schools (0%). Primary barriers to providing physical activity programs were inadequate staffing (54%), funding (39%), and facilities (32%). Girls and low-income youth were identified as the most difficult populations to reach. Directors reported considerable interest in collaborating on interventions to improve youth activity programs. Public parks and recreation centers are understudied settings with the potential for substantial involvement in efforts to promote youth physical activity. PMID- 15358917 TI - Analysis of a database of materials for HIV prevention program evaluation. AB - Program evaluation technology transfer is the transfer of information on program evaluation from research to practitioners. There have been anecdotal reports of a lack of technology transfer materials related to HIV prevention program evaluation, especially materials usable by persons without extensive training in evaluation. The Centers for Disease Control and Prevention reviewed available program evaluation materials relevant to HIV prevention and developed a database of those materials. Materials were classified according to appropriate audience, level of evaluation expertise required, steps in the evaluation process addressed, and other criteria. The database was queried to determine the number of materials available for various combinations of search criteria. These queries revealed that for certain audiences and steps in the evaluation process there are few materials, especially usable by individuals without evaluation experience. The conclusion is that for certain areas of program evaluation, and for certain audiences, more evaluation technology transfer materials are needed. PMID- 15358918 TI - Building a foundation for systems change: increasing access to physical activity programs for older adults. AB - Although 25% of U.S. adults are physically inactive, this percentage increases dramatically for older adults. Organizational change theory guided a state health department in identifying system gaps and developing strategies to expand programming for seniors. A survey of provider agencies in New Jersey assessed (a) capacity for physical activity programs for older adults, (b) accessibility of programs, and (c) barriers to providing programs. One hundred sixty agencies provided physical activity programs to almost 184,000 individuals annually. Fewer than one half of the agencies provided exercise programs for people with disabilities, and only 44% provided in-home programs. Eighty-two percent of program providers wanted to expand programming but cited lack of trained instructors and peer leaders, inadequate facility space, insufficient funding, and limited transportation resources as barriers. Sustaining older adult behavior change requires infrastructure that will ensure access to diverse physical activities. This article provides strategies to expand access to physical activity programs for older adults. PMID- 15358921 TI - Where have all the activists gone? PMID- 15358922 TI - Angels on my shoulders: remarks made to the Association of Nurses in AIDS Care, New York Hilton, November 20, 2003. PMID- 15358923 TI - Stigma: a health barrier for women with HIV/AIDS. AB - The stigma experienced by women with HIV/AIDS is much like that of Hester Prynne in The Scarlet Letter. At the time of diagnosis with HIV/AIDS, women already are aware of the stigma associated with the disease. They immediately see themselves differently and believe others do also. The purpose of this article is to explore the multidimensional effect of stigma on women's efforts to promote, maintain, and enhance their health. The experiences of the women in this study tell of rejection by family members, friends, health care providers, employers, and church members. This rejection caused by stigma affects access to health care, medication adherence, social interaction, and social support. Interventions are needed to decrease the impact of stigma on women with HIV/AIDS so they can achieve a higher level of wellness, increase their life span, continue in the workforce, and improve their quality of life. PMID- 15358924 TI - The patient-provider relationship and adherence to highly active antiretroviral therapy. AB - Four studies completed since 1995 have highlighted the importance of the relationship between the provider and the patient in enhancing adherence behavior. This study extends this work by comparing adherent and nonadherent clients in one high-volume HIV clinic in which the majority of care is provided by nursing staff. The sample comprised 130 clients (108 adherent and 22 nonadherent). Adherence status was determined by clinic staff using established procedures. The indicator of the patient-provider relationship was satisfaction with the care provider as measured within the Patient Satisfaction Questionnaire. Client groups differed significantly on perception of interpersonal manner of care provider (p =.018), care provider conduct total (p <.001), and quality total (p =.017). These findings are consistent with earlier work and underscore the potential importance of the patient-provider relationship as a focus of care for nurses. PMID- 15358925 TI - Substance abuse in HIV-positive women. AB - Early in the U.S. HIV/AIDS pandemic, the role of substance abuse in the spread of AIDS was clearly established. However, the relationship of HIV and substance abuse is more complex than simply noting that injection drug use is a mode of transmission for the virus. HIV infection and substance abuse disorders interact in a complex fashion, with each acting as a potential catalyst or obstacle in the treatment of the other. In this article, we report the results of a qualitative metasynthesis of studies containing information on substance abuse among HIV positive women, using 74 published and unpublished reports. The data on 1,548 women, who were primarily mothers from minority groups, allowed us to construct a trajectory that describes the events of their lives with regard to substance abuse and its intersection with HIV infection. PMID- 15358926 TI - Predictors of virologic success in patients completing a structured antiretroviral adherence program. AB - This study describes the effects of a structured, educational/motivational antiretroviral adherence program on virologic and immunologic parameters in HIV infected patients. Patients were referred because of either self- or provider identified barriers to adherence. All patients completed 6 to 8 weekly sessions with a nurse or adherence counselor, followed by four quarterly sessions. Sessions included an adherence assessment, individualized patient education, review of adherence strategies, motivational messages, anticipatory planning, and adherence tools. The 58 patients had a mean enrollment CD4 (+) count of 223 cells/mm (3) and mean viral load of 196,454 copies/ml. At the last follow-up, the mean CD4 (+) count increased to 308 cells/mm (3) (p <.001), and mean viral load decreased to 43,309 (p <.001). Thirty (51.7%) patients achieved a viral load < 50 copies/ml at any point during follow-up. Factors associated with final viral load < 50 copies/ml included not being a cigarette smoker, receiving lamivudine in one's final regimen, and having an HIV risk behavior other than male-male sex. PMID- 15358927 TI - Looking back to look forward. PMID- 15358928 TI - Social and legal control in China: a comparative perspective. AB - This article exposes the reader to a different social and legal control system and a different way of thinking about crime--that of China. Chinese think positively about the nature of humans and their potential, and China's social control system weaves together a unique combination of formal and informal mechanisms with a strong emphasis on the latter. The criminal justice system constitutes a key element of the social control system, but it appears to be more of a last resort. It is hoped that through a comparative analysis, the reader can develop a deeper understanding of different social and legal systems. PMID- 15358929 TI - School bullying and tackling strategies in Hong Kong. AB - In view of the rising problem of school bullying in Hong Kong, scholars have endeavored to study the prevalence, causes, and tackling strategies of school bullying. This article highlights some typical incidences of school violence and reports research results of school bullying. It is noted unresolved school bullying problems are often a precursor of school violence and delinquency. Based on results from local studies of bullying, this article identifies risk and protective factors that contribute to the emergence and continuation of the bullyvictim problem in Hong Kong. The article argues that suppressive tactics, such as reprimanding bullies, calling parents to school, and suspension, are ineffective. Other than suppressive tactics, adopting a comprehensive antibullying strategy such as assisting students to develop adequate self competency, strong social skills, and good relationships with parents and teachers seems to be a useful antiviolence strategy. PMID- 15358930 TI - Psychosocial characteristics of criminals committing incest and other sex offenses: a survey in a Taiwanese prison. AB - The purpose of this study was to investigate the psychosocial characteristics of criminals who had committed incest or other sexual offenses. The participants, 240 criminals serving sentences for sex offenses in a Taiwanese prison, were divided into two groups: incest offenders (20.4%) and other sex offenders (79.6%). The psychosocial characteristics taken into consideration included age, parental survival, education, marital status, previous crime records, drug and alcohol abuse, diagnosed mental disorders, and victim abuse at the time of the offense. After an analysis of the data, the authors concluded that even though incest offenders showed fewer mental disorders, they needed psychiatric treatment and that this treatment should be focused not only on their mental disorder and related symptoms but especially to correct their abnormal behavior. Also, attention should be given to their psychosocial characteristics. PMID- 15358931 TI - Program-level predictors of antipsychotic medication adherence among parolees. AB - This study examined the impact of three program-level factors (coercion level, type of antipsychotic prescribed, and guardian supervision) on antipsychotic medication adherence among parolees mandated to an outpatient psychiatric clinic. Overall, 70.7% of the participants tested positive for their prescribed antipsychotic. Older patients were more likely to test positive for their prescribed antipsychotic than younger patients, and African American patients (relative to all other race/ethnic groups) were less likely. With regard to program-level factors, perceived coercion was not significantly associated with medication adherence. However, being prescribed an atypical antipsychotic and having a guardian were independently associated with increased adherence, and combined, these two factors were associated with nearly a tenfold increase in the likelihood of patients testing positive for their prescribed antipsychotic agent, relative to patients who were prescribed conventional antipsychotics and were not under guardian supervision. PMID- 15358932 TI - The use of pornography during the commission of sexual offenses. AB - The goal of this study was to examine the use of pornographic materials by sex offenders during the commission of their crimes. A sample of 561 sex offenders was examined. There were 181 offenders against children, 144 offenders against adults, 223 incest offenders, 8 exhibitionists, and 5 miscellaneous cases. All but four cases were men. A total of 96 (17%) offenders had used pornography at the time of their offenses. More offenders against children than against adults used pornography in the offenses. Of the users, 55% showed pornographic materials to their victims and 36% took pictures, mostly of child victims. Nine cases were involved in the distribution of pornography. Results showed that pornography plays only a minor role in the commission of sexual offenses, however the current findings raise a major concern that pornography use in the commission of sexual crimes primarily involved child victims. PMID- 15358933 TI - Pretreatment self-esteem and posttreatment sexual recidivism. AB - The relationship between self-esteem assessed prior to treatment using the Self Esteem Scale and sexual recidivism was explored in two samples of adult male sex offenders, 53 of whom were beginning treatment in the community and 172 of whom were beginning treatment in prison. Sexual reconviction rates were obtained for both samples using a 6-year followup for the community sample and an average follow-up of just less than 4 years for the prison sample. Lower levels of self esteem were associated with higher sexual recidivism rates with similar trends being apparent in both samples. The linear main effect of self-esteem was significant at beyond the .01 level in a logistic regression analysis. Receiver operating characteristics analysis was used to assess the strength of this association and an area-under-the-curve coefficient of .69 was obtained. Results are discussed in terms of their meaning for the relevance of self-esteem as a predictor of recidivism and as a target for treatment. PMID- 15358934 TI - Refusers, dropouts, and completers: measuring sex offender treatment efficacy. AB - A sex offender program delivered in a medium-security prison followed 109 treatment completers and 37 noncompleters for 2 years after release. Noncompleters, those who refused treatment or dropped out, had 6 times the rate of sexual and violent reoffending relative to completers. Among those who completed the program, however, positive evaluations of treatment change, such as quality of disclosure and enhanced victim empathy, found in posttreatment assessments did not correlate with recidivism. Furthermore, completers did not differ in their rates of recidivism from pretreatment rates predicted by the Static 99, an actuarial measure of anticipated sexual and violent recidivism. We conclude that the program did not influence propensities for sexual and violent recidivism but rather served as a prolonged screening instrument for sex offenders whose failure to comply with treatment attendance predicted higher rates of recidivism. PMID- 15358935 TI - Urban and rural differences in the relationship between substance use and violence. AB - This article examines the relationship between substance use and violence across rural-urban and Appalachian places of residence. The data come from a larger study on the health service use of 637 men who have a history of chronic substance abuse and who were incarcerated in four Kentucky prisons. The findings generally support previous research on substance use and violence but do not support Fischer's (1995) subculture theory of urbanism. Contrary to expectations, the population size of the prisoners' residence was not significantly associated with the prisoners' levels of violent victimization, violence toward others, violence toward intimate partners, or overall violence in the year prior to incarceration. Appalachian residency was also not associated with violence. Recognizing that the effect of substance use on violence perpetrated against others does not vary significantly by urban or rural residence may be helpful for designing violence prevention programs and planning law enforcement efforts. PMID- 15358936 TI - Post-traumatic stress disorder in maltreated youth: a study of diagnostic comorbidity and child factors. AB - The study of post-traumatic stress disorder (PTSD) in maltreated youth has received increased attention, though extensive comparisons to maltreated youth without PTSD and administrations of anxiety-based structured diagnostic interviews remain needed. We examined maltreated youth with or without PTSD using structured diagnostic interviews and standardized child self-report measures. We hypothesized that maltreated youth with PTSD, compared to their peers without PTSD, would experience significantly greater duration of abuse, diagnostic comorbidity, PTSD symptomatology, dysfunctional family environment, and avoidant coping styles. Results indicated that the group with PTSD did indeed experience significantly greater duration of abuse, diagnostic comorbidity, and PTSD symptomatology, though less so dysfunctional family environment or avoidant coping styles. The presence of a mood or anxiety disorder was highly predictive of PTSD in this sample. Results are discussed within the context of evolving etiological and maintenance models of PTSD in maltreated youth. PMID- 15358937 TI - Adult psychopathology and intimate partner violence among survivors of childhood maltreatment. AB - Childhood maltreatment is associated with psychopathology and revictimization in adulthood. Whether different types of childhood maltreatment have different long term consequences, however, is largely unknown. The participants in this study included 42 female victims of intimate partner violence and 30 women with no history of serious trauma. These women completed measures of their trauma history, current psychological symptoms, and severity of current abuse. Regression-based techniques were used to assess the relationships among these constructs. Childhood maltreatment was associated with current distress and current conflict in relationships. There was a positive relationship between number of types of childhood traumatization and psychopathology. Symptoms of post traumatic stress disorder (PTSD) and depression were negatively associated with current relationships involving physical assault. This work underscores the importance of assessing multiple types of childhood maltreatment and of considering the cumulative effects of such experiences. PMID- 15358938 TI - Dissociation and variability of adult attachment dimensions and early maladaptive schemas in sexual and violent offenders. AB - The aim of this study was to investigate the presence of constructs that may indicate the presence of disorganized attachment style in sexual and violent offenders. Constructs measured were dissociation, variability on self-report measures of attachment style and early maladaptive schemas, and variability in observed behavior. Data on variability was collected at four time intervals, approximately 3 weeks apart. No differences between the groups were found in variability of self-reported attachment style. Both offending groups showed greater variability in early schemas and higher levels of dissociation than the nonoffending group. Sex offenders showed greater variability than violent offenders in behaviors related to distress. PMID- 15358939 TI - The long-term effects of child sexual abuse by female perpetrators: a qualitative study of male and female victims. AB - Although the long-term effects of sexual abuse by men have been studied extensively, minimal research has explored the effects of sexual abuse by women. This qualitative study explores the experience and long-term impact of sexual abuse by women. The data were derived from in-depth interviews with 14 adult victims (7 men, 7 women) of child sexual abuse by females. Most respondents reported severe sexual abuse by their mothers. The vast majority of participants reported that the experience of female-perpetrated sexual abuse was harmful and damaging. As a result of the sexual abuse, male and female respondents reported long-term difficulties with substance abuse, self-injury, suicide, depression, rage, strained relationships with women, self-concept and identity issues, and a discomfort with sex. In light of the popular and professional perceptions that sexual abuse by women is relatively harmless as compared to sexual abuse by men, the implications of these long-term effects are discussed, particularly in relation to professionals working in the area of child sexual abuse. PMID- 15358940 TI - The impact of past sexual experiences on attributions of responsibility for rape. AB - Two factors potentially affect observers'attributions of responsibility to a rape survivor: how closely they identify with the survivor and how much they adhere to rape myths. To assess the impact of these factors, 157 female college students categorized by their sexual assault history and by their acceptance of rape myths, evaluated a sexual assault scenario. It was hypothesized that previously victimized participants would attribute less blame, and that those participants who self-identify as rape survivors would view the assault as rape more often than other participants. Neither hypothesis was supported. As expected, those who scored higher on the Rape Myths Acceptance Scale(RMAS) blamed the victim more and were less likely to believe rape had occurred. Only one subscale of the RMAS, Adversarial Sexual Beliefs, was consistently associated with participants'evaluation of the scenario, suggesting that this dimension may be especially important to understanding an observer's assessment of rape. PMID- 15358941 TI - Cross-validation of the Self-Appraisal Questionnaire (SAQ): an offender risk and need assessment measure on Australian, British, Canadian, Singaporean, and American offenders. AB - The Self-Appraisal Questionnaire (SAQ) is a 72-item self-report measure designed to predict violent and nonviolent recidivism among adult criminal offenders. The results from using samples from Australia, Canada, England, Singapore, and two samples from the United States (North Carolina and Pennsylvania) indicated that (a) the SAQ has sound psychometric properties, with acceptable reliability and concurrent validity for assessing recidivism and institutional adjustment; (b) there were no significant differences among the scores of the White, African American, Hispanic, and Aboriginal Australian offenders on the SAQ; (c) there were no significant differences among offenders who completed the SAQ for research purposes versus offenders who completed it as part of a decision-making process. Results provided support for the validity of the SAQ to be used with the culturally diverse offenders involved in this research and provided further evidence that contradicts concerns that the SAQ as a self-report measure may be susceptible to lying, and self-presentation biases. PMID- 15358942 TI - A comparison of nonviolent, psychologically violent, and physically violent male college daters. AB - This brief report explores dating violence by comparing three groups of male college students (nonabusive, psychologically abusive only, and physically abusive). These men were compared on measures of impulsivity, problems with alcohol, life satisfaction, anger management skills, history of witnessing abuse, history of experiencing abuse, and relationship satisfaction. Data for this analysis were obtained from a sample of 115 male college students. Differences between the three groups of men were found in the levels of problems with alcohol, relationship satisfaction, and anger management skills. Anger management skills best differentiated the three groups of men leading to the conclusion that dating violence prevention and intervention strategies with male college students should address anger management skills. PMID- 15358943 TI - Extracorporeal life support in pediatric and neonatal critical care: a review. AB - Extracorporeal life support (ECLS) is a modified form of cardiopulmonary bypass used to provide prolonged tissue oxygen delivery in patients with respiratory and/or cardiac failure. The first large-scale success of ECLS was achieved in the management of term newborns with respiratory failure. ECLS has become an accepted therapeutic modality for neonates, children, and adults who have failed conventional therapy and in whom cardiac and/or respiratory insufficiency is potentially reversible. The use of ECLS allows one to reduce other cardiopulmonary supports and apply a gentle ventilation strategy in a population of severely compromised critical care patients. ECLS has now been employed in more than 26,000 neonatal and pediatric patients with an overall survival rate of 68%. ECLS has evolved significantly over 25 years of clinical practice; patient selection for this complex and highly invasive therapy, as well as how ECLS is employed in different patient groups, is constantly changing. Generally, ECLS is used more liberally now than in the past. The number of patients requiring this support, however, is declining yearly, and those patients who receive ECLS compose a more severe subset of an intensive care population. This review provides an overview of the development of ECLS and the equipment and techniques employed. The use of ECLS for neonatal respiratory failure, pediatric respiratory failure, and cardiac support are outlined. Management of the ECLS patient is discussed in detail, and outcome of these patients is reviewed. Finally, current trends and future implications of ECLS in neonatal and pediatric critical care are addressed. PMID- 15358944 TI - Venomous snakebites worldwide with a focus on the Australia-Pacific region: current management and controversies. AB - Snakebites are estimated to cause approximately 100,000 deaths each year worldwide and disproportionately affect rural populations in resource-poor settings. Snake venoms may produce local tissue damage and/or distinct clinical syndromes, including neurotoxicity, coagulopathy, hypotension, rhabdomyolysis, and renal failure. Field management is aimed at delaying systemic absorption of toxins, minimizing local damage and infection, and expediting transport to medical facilities. The use of the pressure-immobilization method remains controversial. The use of antivenom, administered in a timely fashion and in adequate doses, is the mainstay of hospital treatment of significant envenomation. The availability, efficacy, and safety of antivenoms vary throughout the world, with a current crisis in antivenom supplies. PMID- 15358945 TI - Recombinant factor VIIa to control excessive bleeding following surgery for congenital heart disease in pediatric patients. AB - The purpose of this article is to evaluate the efficacy of recombinant factor VII (rFVIIa) in the treatment of bleeding following cardiac surgery in a pediatric population. The study included a case series of postcardiac surgical patients with chest tube output of = 4 mL/kg/h for the initial 3 postoperative hours who received rFVIIa. Chest tube output for the 3 hours before and the 3 hours after rFVIIa was compared using a paired t test. In addition, chest tube output for the initial 3 postoperative hours and the 3 hours following rFVIIa was compared to 8 control patients who did not require rFVIIa. Recombinant factor VIIa was administered to 9 children (age = 9 +/- 4 years) following repair of tetralogy of Fallot (6), closure of ventricular septal defect (1), closure of sinus venosus atrial septal defect (1), and mitral valve repair (1). Chest tube output for the initial 3 postoperative hours prior to the administration of rFVIIa was 5.8 +/- 2.8 mL/kg/h and decreased to 2.0 +/- 1.3 mL/kg/h for the 3 hours following the administration of rFVIIa (P =.002). In the patients that did not receive rFVIIa, chest tube output for the first 3 postoperative hours was 1.6 +/- 0.9 mL/kg/h and 1.2 +/- 0.6 mL/kg/h for the next 3 hours (P = nonsignificant when compared to chest tube output for the 3 hours following rFVIIa in patients who received rFVIIa). No adverse effects were noted. Recombinant factor VIIa decreased chest tubing bleeding following cardiac surgery in children. Given its potential therapeutic impact, rFVIIa warrants further investigation in the pediatric cardiac population. PMID- 15358946 TI - Protocol-driven ventilator management in children: comparison to nonprotocol care. AB - The purpose of this study was to compare ventilator weaning time, time to spontaneous breathing, and overall ventilator hours duration with use of a ventilator management protocol (VMP) versus standard nonprotocol-based care in a pediatric intensive care unit. A multidisciplinary task force developed a comprehensive protocol for ventilator management with four specific phases: initial ventilator set up and adjustment, weaning, minimal settings, and spontaneous mode prior to extubation. Medical records of ventilated patients both before and after protocol implementation were reviewed. A total of 187 patients were studied (89 nonprotocol and 98 VMP patients). No differences were seen between groups in PRISM scores, Murray scores, or oxygenation indices, but VMP patients were significantly younger (P =.03). Ventilator weaning times (P =.005) and time to spontaneous breathing modes (P =.006) were significantly decreased in VMP patients compared to nonprotocol patients, but overall ventilator duration was not significantly different. No significant differences were seen in extubation failure, use of corticosteroids, or use of racemic epinephrine between groups. Use of an institution-specific VMP developed by a multidisciplinary team was associated with significantly reduced ventilator weaning time and time to spontaneous breathing. Further studies are needed. PMID- 15358947 TI - The occurrence of posttraumatic stress disorder in patients following intensive care treatment: a cross-sectional study in a random sample. AB - Posttraumatic stress disorder (PTSD) can now be applied to patients who have just had intensive care treatment. The authors tested whether the reliability of the diagnosis depends on the measuring instrument used and whether the occurrence of PTSD is related to the presence of preexisting psychic disorders or to a worsening of the patients' general condition. The authors examined 41 former patients of an intensive care unit (ICU) with the Posttraumatic Scale (PTSS-10) and, afterward, with the Structured Clinical Interview (SCID). Screening with PTSS-10 showed that 17.07% of the patients met the recommended threshold for probable diagnosis of PTSD of 35 or more points. With SCID, PTSD could be confirmed in 9.76% of the cases. Patients with a prior psychiatric disease showed more frequent (P < .02) occurrences of PTSD. The severity of their Acute Physiology and Chronic Health Evaluation System II (Apache II) scores had no significant influence. PTSS-10 can be used for a preliminary diagnostic orientation. With the PTSD module of the SCID, a structured diagnostic tool is available. Almost twice as many PTSD cases were diagnosed among the subjects with the PTSS-10 as with SCID. Post-ICU PTSD occurred more frequently among patients with prior psychiatric diseases; however, it was not dependent on exacerbation of the patient's Apache II scores. PMID- 15358948 TI - Pulmonary artery rupture induced by a pulmonary artery catheter: a case report and review of the literature. AB - Placement and use of pulmonary artery catheters (PACs) carry potential risks. The authors describe a case of a patient who developed massive hemoptysis after placement of a PAC that caused a rupture of the pulmonary artery with pseudoaneurysm formation. Treatment was successfully achieved with transcatheter coil embolization. Pulmonary artery rupture and pseudoaneurysm formation are among the most serious complications of PAC use because of the associated risk of mortality. Patients with this complication may be asymptomatic or may present with variable amounts of hemoptysis immediately or days after using a PAC. The gold standard diagnostic test is pulmonary angiography, and the treatment of choice for most patients is transcatheter embolization. Physicians and other health care personnel handling these catheters should be familiar with the specific PAC balloon's inflation limits to avoid complications that may injure the patient. PMID- 15358949 TI - Magnetization transfer magnetic resonance imaging in the assessment of neurological diseases. AB - In several white matter diseases of the central nervous system (CNS), and in particular in multiple sclerosis, conventional magnetic resonance imaging (MRI) has proved to be sensitive for detecting lesions and their changes over time. However, conventional MRI is not able to characterize and quantify the tissue damage within and outside such lesions. Magnetization transfer MRI (MT-MRI) is a quantitative MRI technique with the potential to overcome this limitation and, as a consequence, to provide additional information about the nature and the extent of tissue damage. Metrics derived from MT-MRI can quantify the structural changes occurring within and outside lesions visible on conventional MRI scans. The present review summarizes the major contributions given by MT-MRI to provide an accurate in vivo picture of the heterogeneity of CNS pathology and, ultimately, to improve our ability to monitor the evolution of various neurological conditions. PMID- 15358950 TI - The accuracy of transcranial Doppler in the diagnosis of stenosis or occlusion of the terminal internal carotid artery. AB - BACKGROUND AND PURPOSE: Transcranial Doppler (TCD) can detect intracranial stenoses and occlusions that can help in the diagnosis and management of ischemic stroke. The accuracy parameters for lesions located in the terminal internal carotid artery (TICA) are less known, unlike other basal cerebral vessels. PATIENTS AND METHODS: The authors studied consecutive patients referred for TCD who underwent contrast angiography or magnetic resonance angiography. They calculated the sensitivity, specificity, positive and negative predictive values (PPV and NPV), and likelihood ratios. RESULTS: Forty-three patients had TCD and angiography: mean age was 57 +/- 20 years, and 65% were men. Twenty-two patients were diagnosed with TICA stenosis or occlusion on TCD underwent angiography. Four patients had abnormal TCD findings that were not confirmed by angiography. Two of 21 patients with normal TCD showed moderate (< 50%) stenosis of the TICA and cavernous segment of the internal carotid artery at angiography. Accuracy parameters for TCD were as follows: sensitivity = 90% (confidence interval [CI], 63%-96%), specificity = 83% (CI, 61%-94%), PPV = 82%, NPV = 86%, positive likelihood ratio = 5, and negative likelihood ratio = 0.17. CONCLUSIONS: TCD is a sensitive screening tool for the lesions in the TICA. Specificity is likely affected by a wide spectrum of the stenosis severity shown at angiography and time lags between the studies. PMID- 15358951 TI - Diagnosis of moyamoya disease with transcranial Doppler sonography: correlation study with magnetic resonance angiography. AB - BACKGROUND AND PURPOSE: Although the diagnosis of moyamoya disease may be confirmed by digital subtraction angiography, recent studies have shown the accuracy of magnetic resonance angiography. Characteristics of transcranial Doppler, a noninvasive and cost-effective method, and specific transcranial Doppler parameters reflecting distinct vascular status in moyamoya disease are explored. METHOD AND PATIENTS: Consecutive patients (> 15 years of age) diagnosed with moyamoya disease by a typical clinical history and digital sub traction angiography or magnetic resonance angiography were included. The statuses of the anterior, middle, and posterior cerebral arteries were graded as stage 1, stage 2, and stage 3 by magnetic resonance angiography. Mean flow velocity and pulsatility index were compared between these groups, and the receiver operating characteristic analysis was used to define transcranial Doppler criteria for distinct vascular status. RESULTS: Forty-five patients were included (37 women; mean age, 34.9 +/- 11.4 years). Mean flow velocity was higher and pulsatility index was lower in stage 2 (P < .01), while mean flow velocity was lower and pulsatility index was higher in stage 3 than in stage 1 (P < .01). Cutoff values reflecting stenosis or occlusion with substantial sensitivity and specificity were as follows: mean flow velocity > 85 cm/s or pulsatility index < 0.60 for stage 2, and mean flow velocity < 50 cm/s for stage 3 of middle cerebral artery; mean flow velocity > 80 cm/s or pulsatility index < 0.60 for stage 2 of anterior cerebral artery; and mean flow velocity > 60 cm/s or pulsatility index < 0.60 for stage 2 of posterior cerebral artery. CONCLUSION: Transcranial Doppler may help to refine magnetic resonance angiography findings and thus help clinicians differentiate severity or stages of moyamoya disease. PMID- 15358952 TI - Ictal single-photon emission computed tomography imaging in extra temporal lobe epilepsy using statistical parametric mapping. AB - PURPOSE: To examine the application of statistical parametric mapping (SPM) to analyze ictal single-photon emission computed tomography (SPECT) scans in surgical candidates with extratemporal lobe epilepsy. METHODS: The authors selected patients who underwent successful ictal SPECT acquisition in the process of surgical treatment of intractable partial epilepsy. Thirteen patients were identified who met inclusion criteria for confident seizure localization from either intracranial electroencephalogram recordings or epilepsy surgery outcome. In these cases, ictal scans were registered to an in-house-developed normal SPECT atlas composed of 14 spatially normalized brains of normal subjects. SPM96 was used to test on a voxel-by-voxel basis for statistically significant increases in blood flow associated with each patient's ictal scan. The results were then mapped back onto the patient's magnetic resonance image (MRI) for final interpretation. Statistical parametric mapping (SPM) analysis of ictal SPECT scans was compared to both conventional visual interpretation and the analysis of subtraction ictal SPECT co-registered to MRI (SISCOM). RESULTS: Ten of 13 patient scans showed localizing focal ictal increases in regional cerebral blood flow, all of which were concordant with ultimate epilepsy localization. Of the 3 cases not localized with SPM, 1 was localized by conventional visual interpretation and another, not localized by visual interpretation, was correctly localized with SISCOM. Two cases not localized by SISCOM were localized by both visual and SPM analysis. CONCLUSIONS: This work provides supportive evidence for proof of principle that SPM can be used to provide objective, accurate analysis of ictal SPECT scans in patients with extratemporal lobe epilepsy. PMID- 15358953 TI - Transcranial Doppler in asymptomatic carotid stenosis representing hemodynamic impairment: correlation study with magnetic resonance imaging. AB - BACKGROUND AND PURPOSE: Since the benefit of an endarterectomy in an asymptomatic carotid stenosis is not as clear as that in symptomatic stenosis, identifying patients with a high risk of stroke will allow the improved selection of surgical candidates. To evaluate the role of transcranial Doppler (TCD) in assessing an asymptomatic stenosis, this study examined the association of a TCD-detected hemodynamic change with a stenosis-related abnormality on magnetic resonance imaging (MRI). METHODS: Asymptomatic patients with a more than 60% internal carotid artery stenosis were enrolled in this study. The hemodynamic change in the TCD was analyzed in terms of the demographic characteristics, the stroke risk factors, and the stenosis-related MRI abnormalities that are a proven risk factor of a further stroke. RESULTS: Fifty-three patients were retrospectively recruited. While there were no differences in the demographic characteristics or frequency of the stroke risk factors, the proportion of patients with stenosis related MRI abnormality was significantly higher in patients with a hemodynamic change on TCD (13/19 vs 6/34, P < .01; odds ratio = 10.11; 95% confidence interval = 2.73-37.4). CONCLUSIONS: The TCD-detected hemodynamic changes may provide useful information for assessing the frequency of silent stroke in patients with an asymptomatic carotid stenosis. PMID- 15358954 TI - Three-dimensional contrast-enhanced magnetic resonance angiography of carotid artery after stenting. AB - PURPOSE: This study presents 3-dimensional, contrast-enhanced, magnetic resonance (MR) angiographic appearances of the carotid artery after stenting. METHODS: The authors reviewed contrast-enhanced MR angiograms of 18 carotid arteries in 13 patients. These arteries were not stenotic or occluded in the stented segment, as shown on carotid angio-grams (11 patients) or carotid Doppler images (2 patients) obtained later. RESULTS: Five patients had 1 stent in each of their bilateral carotid arteries. One patient with 1 stent in 1 artery underwent the examination twice with different MR coils. Different artifact patterns were found in 19 carotid-stent examinations. Type 1a involved short segments of false narrowing or false focal occlusion at both ends of the stent (n = 12); the carotid artery in the stented segment between the ends was normal. Types 2a and 2b occurred along the stented segment of the artery between the 2 ends of the stent. Type 2a was milder, with segmental faint signal intensity or luminal stenosis in the entire length of the stented segment (n = 5). Type 2b was total loss of luminal signal intensity over the whole length of the stented segment (n = 2). Last, type 1b was a band of hyperintensity at the ends of type 2a or 2b artifacts. Types 1a and 1b were due to susceptibility effect, and types 2a and 2b were from radio-frequency shielding effect. Both of these effects were identified in MR angiogram of nitinol stents and stainless steel stents. CONCLUSION: Awareness of these artifacts is essential to prevent misdiagnosis. PMID- 15358955 TI - Power m-mode transcranial Doppler for diagnosis of patent foramen ovale and assessing transcatheter closure. AB - BACKGROUND AND PURPOSE: Transcatheter closure of patent foramen ovale (PFO) can benefit from a less invasive diagnostic method than transesophageal echocardiography (TEE). Thirty-three gate power m-mode transcranial Doppler (pmTCD) was evaluated for its accuracy in diagnosis of PFO and utility in evaluating residual intracardiac right-to-left shunt (RLS) following transcatheter closure. METHODS: The sensitivity of pmTCD and single-gate TCD (sgTCD) to detect contrast bubble emboli through RLS was compared during transcatheter PFO closure. During 100 preclosure diagnostic evaluations and in 81 postclosure assessments, embolic tracks on pmTCD were counted following intravenous contrast injections and were graded using a 6-level logarithmic scale. The accuracy of TEE and pmTCD was separately compared to PFO anatomical findings during transcatheter closures. RESULTS: There were significantly more microemboli detectable on pmTCD (322 +/- 166; 95% confidence interval [CI], 388 257) than on sgTCD (186 +/- 109; 95% CI, 229-143; P < .001). McNemar change tests suggest that the diagnostic capabilities of pmTCD and TEE for detecting PFO are comparable and correspond to the anatomical findings determined during cardiac catheterization (P = .69 and .45, respectively). During 6-month postclosure evaluation (mean = 185 days), 66% of the patients demonstrated successful closure without significant RLS (ie, grades 0, I, or II), and 34% were found to have incomplete closure with significant RLS (ie, grades III, IV, or V). CONCLUSIONS: pmTCD provides greater sensitivity to contrast bubble emboli than does sgTCD. Among candidates for transcatheter closure, pmTCD provides an improved noninvasive method for diagnosing PFO and evaluating transcatheter closure. PMID- 15358956 TI - Changes in the heterogeneity of cerebral glucose metabolism with healthy aging: quantitative assessment by fractal analysis. AB - BACKGROUND AND PURPOSE: It has been shown that heterogeneity of cerebral glucose metabolism is increased in neuropsychiatric degenerative diseases. However, proper assessment of older patients requires knowledge about the effect of aging on heterogeneity. This study characterized the effects of aging on the heterogeneity of the distribution of cerebral glucose metabolism in healthy volunteers. METHODS: Sixty-six healthy volunteers (age range, 19-75 years) underwent flurodeoxyglucose brain positron emission tomography (PET), and all the PET images were spatially normalized onto a previously segmented standard brain template to parcel the brain regions automatically. Fractal dimension was regarded as a quantitative measurement for the heterogeneity of cerebral glucose metabolism and obtained for 9 brain regions. Participants were subdivided into young/midlife and elderly groups, and the Student t test was applied to the comparison of fractal dimensions in those groups. Analysis of covariance was performed for each region to explore the effects of age, gender, age-by-gender interaction, and total counts in the brain on the observed metabolic heterogeneity. RESULTS: Fractal dimensions were higher for elderly volunteers in most brain regions. Differences between the 2 groups in fractal dimension emerged within the whole gray matter, temporal lobe, striatum, and cingulate. No significant gender differences, age-by-gender interactions, or total counts were observed. Significant age effects were observed in the whole gray matter, frontal lobe, temporal lobe, striatum, and cingulate gyrus. CONCLUSIONS: Heterogeneity in the cerebral glucose metabolism of healthy volunteers increased with age, and individual variations of heterogeneity were higher in older volunteers. However, there was no significant difference between male and female volunteers of the same age. The effect of age on heterogeneity was not regionally uniform. PMID- 15358957 TI - Contralateral cerebral blood flow velocity changes after intracarotid amobarbital injection. AB - BACKGROUND AND PURPOSE: The intracarotid amobarbital procedure (IAP) leads to a prompt decrease in ipsilateral middle cerebral artery (MCA) mean blood flow velocity (MFV). Little is known about contralateral MFV changes. METHODS: The authors investigated bilateral MCA MFV using transcranial Doppler sonography (TCD) in 8 patients with epilepsy undergoing IAP. Measurements were excluded from analysis if angiography revealed any signs of interhemispheric cross-flow. RESULTS: With in 64 seconds after amobarbital injection, ipsilateral MFV decreased to a mean of 44.4% +/- 7.5% of baseline value (P < .01). In the absence of interhemispheric cross-flow and within 68 seconds, contralateral MFV decreased to 83.1% +/- 7.9% (P < .01). CONCLUSIONS: The observed decrease of contralateral MFV was not caused by amobarbital cross-perfusion. A possible underlying mechanism may be interhemispheric deafferentation. PMID- 15358958 TI - Spontaneous hyperacute postischemic hemorrhage leading to death. AB - Hyperacute, spontaneous, and severe parenchymal postischemic hemorrhage is considered rare but might be frequently misdiagnosed as primary intracerebral hemorrhage. The authors report 2 patients with catastrophic postischemic hemorrhage unrelated to anticoagulation, thrombolytics, or coagulopathy. Patient 1 was a 73-year-old woman with left posterior frontal lobe infarction, followed at 5.5 hours by massive postischemic hemorrhage leading to death. Patient 2 was a 52-year-old man with recent brain irradiation who developed left middle cerebral artery occlusion and basal ganglia infarction. Initial gradient-echo magnetic resonance imaging (MRI) showed petechial hemorrhage within areas of infarction and widespread old microbleeds. He developed massive postischemic hemorrhage at 12 hours, leading to death. The authors conclude that early brain imaging is essential to distinguish postischemic from primary brain hemorrhage. In patients with ischemic stroke, MRI findings such as hyperacute petechial hemorrhage or chronic cerebral microbleeds might predict subsequent parenchymal hemorrhage and thus have implications for tissue plasminogen activator use. PMID- 15358959 TI - Simple partial status epilepticus localized by single-photon emission computed tomography subtraction in chronic cerebral paragonimiasis. AB - A patient with chronic cerebral paragonimiasis began to have new motor seizures of the right face manifested by clonic contractions that occurred several hundred times a day, consistent with simple partial status epilepticus. Ictal electroencephalogram discharges started from the left frontal region and then spread to the left hemisphere with left frontal maximum. But clinical seizures were limited to the right face. The frequent partial seizures were controlled by the intravenous infusion of phenytoin. Brain magnetic resonance imaging showed multiple conglomerated round nodules with encephalomalacia in the left temporal and occipital lobes. Applying the technique of ictal-interictal single-photo emission computed tomography subtraction, the authors were able to localize the focal ictal-hyperperfusion on left precentral cortex adjacent to the lesions that correspond to the anatomical distribution of left face motor area. PMID- 15358960 TI - Reversible cerebellar lesions induced by metronidazole therapy for helicobacter pylori. AB - Metronidazole is widely used for chronic or refractory infection and has recently also been used for the treatment of Helicobacter pylori. The authors report the case of a Japanese patient presenting with reversible cerebellar lesions induced by prolonged administration of metronidazole for treatment of H pylori with magnetic resonance imaging findings. Although rare, prolonged and high-dose administration of metronidazole may induce cerebellar lesions. Increased awareness of this phenomenon is important, as these lesions are reversible with discontinuation of this drug. PMID- 15358961 TI - Novel neuroimaging findings in a patient with cerebral Whipple's disease: a magnetic resonance imaging and positron emission tomography study. AB - The authors report a 43-year-old patient with histopathologically proven cerebral Whipple's disease. Magnetic resonance imaging (MRI) revealed a multilayered left frontal lesion without mass effect, no perifocal brain edema, no contrast enhancement, and a thin shell of fluid signal that presented as an incomplete, open ring. An [11C]methionine positron emission tomography (PET) study showed low uptake below the threshold that is characteristic for brain tumors. In precise co registration to the MR images, the PET data showed that increased uptake was mainly located in the direct adjacent part of the MRI lesion. The fluid signal on MRI corresponded to the extensive outflow of fluid from the lesion, which was observed during neurosurgical resection, and also to the neuropathological findings. The authors conclude that this cerebral manifestation of Whipple's disease made a unique and hitherto undescribed appearance on MRI; uptake pattern of PET amino acid tracer may help in the preoperative distinction of inflammatory from neoplastic lesions. PMID- 15358962 TI - Proton magnetic resonance spectroscopy and magnetic resonance imaging findings in a patient with central nervous system paracoccidioidomycosis. AB - The authors describe a patient with brain paracoccidioidomycosis whose magnetic resonance image (MRI) showed multiple hypointense lesions on T2-weighted images with peripheral enhancement after gadolinium injection. Single-voxel proton magnetic resonance spectroscopy (1H-MRS) of one of the lesions showed 2 peaks at 0.9 and 1.32 ppm, corresponding to lipid signals, indicating intense necrosis. The other characteristic peaks of 1H-MRS were undifferentiated from baseline. These findings, although not specific, may help to differentiate fungal abscess from tumoral lesions and other types of abscess. PMID- 15358963 TI - Anatomical functional changes in a patient presenting a complex malformation of cortical development. AB - The authors describe a case of right fronto-parietal micropoligyria associated with small schizencephaly clefts and the presence of a frontal open-lip schizencephaly with corpus callosum agenesis. A functional magnetic resonance imaging (fMRI) study was performed to evaluate the possible reorganization of cortical functions in a patient presenting a complex malformation pattern and to investigate which cortical areas were activated during left finger movements. An fMRI study was performed during the execution of a repetitive index finger-to thumb opposition movement with the right hand and the left hand in 2 separate sessions. Movement of the right hand induced a normal motor activation pattern involving the contralateral left sensory-motor cortex. Movement of the left hand produced significant activation of brain cortex. This fMRI study highlights the compensatory role of the ipsilateral cortical pathways in hand movements in the case of a complex brain malformation that involves the main motor activation areas. PMID- 15358964 TI - Unsuccessful tissue plasminogen activator treatment of acute stroke caused by a calcific embolus. AB - Intravenous (IV) administration of tissue plasminogen activator (tPA) given to patients during acute cerebral ischemia according to National Institute of Neurological Disorders and Stroke (NINDS) guidelines improves clinical outcome by 11% to 14%. The success of IV tPA stroke therapy is dependent on several previously reported factors. The authors suggest that the presence of calcification within an embolus may represent an additional important factor. This report describes a patient with an acute stroke secondary to a spontaneous calcific cerebral embolus who had a negative outcome despite receiving proper thrombolytic therapy. PMID- 15358969 TI - Two decades of organizational change in health care: what have we learned? AB - The 1980s and 1990s witnessed a substantial wave of organizational restructuring among hospitals and physicians, as health providers rethought their organizational roles given perceived market imperatives. Mergers, acquisitions, internal restructuring, and new interorganizational relationships occurred at a record pace. Matching this was a large wave of study and discourse among health services researchers, industry experts, and consultants to understand the causes and consequences of organizational change. In many cases, this literature provides mixed signals about what was accomplished through these organizational efforts. The purpose of this review is to synthesize this diverse literature. This review examines studies of horizontal consolidation and integration of hospitals, horizontal consolidation and integration of physician organizations, and integration and relationship development between physicians and hospitals. In all, around 100 studies were examined to assess what was learned through two decades of research on organizational change in health care. PMID- 15358970 TI - Primary care outcomes in patients treated by nurse practitioners or physicians: two-year follow-up. AB - This study reports results of the 2-year follow-up phase of a randomized study comparing outcomes of patients assigned to a nurse practitioner or a physician primary care practice. In the sample of 406 adults, no differences were found between the groups in health status, disease-specific physiologic measures, satisfaction or use of specialist, emergency room or inpatient services. Physician patients averaged more primary care visits than nurse practitioner patients. The results are consistent with the 6-month findings and with a growing body of evidence that the quality of primary care delivered by nurse practitioners is equivalent to that by physicians. PMID- 15358971 TI - What matters to low-income patients in ambulatory care facilities? AB - Poor, uninsured, and minority patients depend disproportionately on hospital outpatient departments (OPDs) and freestanding health centers for ambulatory care. These providers confront significant challenges, including limited resources, greater demand for services, and the need to improve quality and patient satisfaction. The authors use a survey of patients in OPDs and health centers in New York City to determine which aspects of the ambulatory care visit have the greatest influence on patients' overall site evaluation. The personal interaction between patients and physicians, provider continuity, and the general cleanliness/appearance of the facility stand out as high priorities. Access to services and interactions with other facility staff are of significant, although lesser, importance. These findings suggest ways to restructure the delivery of care so that it is more responsive to the concerns of low-income patients. PMID- 15358972 TI - Severity of illness and ambulatory care-sensitive conditions. AB - This study describes how severity of illness may refine the definition of ambulatory care-sensitive conditions, or ACSCs. Hospital discharge abstract data from Philadelphia were combined with census data to develop population-based adjusted rates of hospitalization for diabetes and asthma, two ACSCs. By stratifying ACSC hospitalization by severity of illness, variations were observed by age, race, and gender. Minority groups may be at higher risk for less access to outpatient primary care and were observed to have higher rates of more severely ill, Stage 3 hospitalization. Geographic map displays indicated wide ranges of age-sex-adjusted rates for high-severity hospitalization in the five county Philadelphia region. This refined ACSC measure may help to identify specific groups and clinical conditions within a population to assist health care planners estimate health care resources such as facilities, manpower, and programs, as well as to evaluate their outcomes. PMID- 15358973 TI - How health policy and health services researchers are compensated: analysis of a nationwide salary survey. AB - The membership of Academy Health, a professional organization, was invited to complete an anonymous Web-based survey in 2002. Responses were received from 1,140 of 2,633 surveyed (43 percent). Fifty-six percent worked in academic institutions or teaching hospitals, 34 percent in the private sector or foundations, and 10 percent in government. Most (96 percent) had at least one advanced degree, and the diversity of educational backgrounds was pronounced. The median annual salary was $99,000. Salaries were highest in the private sector, followed by academic and government settings. There were large regional variations, with higher salaries in the Mid-Atlantic and New England regions. Adjusted data suggested these higher regional salaries were inadequate to compensate for higher local cost of living. Among academic respondents, nonadjusted salaries increased with advancing faculty job titles, but this seniority effect was inconsistent across geographic regions. Junior faculty salaries, when adjusted for cost of living, were more similar across regions than salaries at the full professor level. PMID- 15358974 TI - An exceptional case of plagiarism! PMID- 15358975 TI - Evidence-based guidelines for the pharmacological management of substance misuse, addiction and comorbidity: recommendations from the British Association for Psychopharmacology. PMID- 15358976 TI - Gender differences and the effects of ketamine in healthy volunteers. AB - We present a critical perspective of the impact of gender differences on a widely accepted model of ketamine psychosis in healthy volunteers. Male and female patients with schizophrenia present with different symptomatology, disease course and response to pharmacological intervention. Accordingly, it is expected that ketamine psychosis in healthy volunteers fulfils this face validity. Pre-clinical studies in rats indicate a gender difference in response to ketamine administration. However, a review of the literature to date indicates that studies carried out in healthy volunteers have used all male or mixed samples, indicating the need for further studies comparing the psychopathological effects of ketamine in males and females. PMID- 15358977 TI - Receptor mechanisms in the treatment of schizophrenia. AB - There remain many limitations to the treatment of schizophrenia. In addition to the poor response of negative and cognitive symptoms to antipsychotics, and the substantial proportion of poor- or non-responders, there are a variety of unpleasant and restricting side-effects of these drugs. The introduction of several 'atypical' drugs, with diminished propensity to cause extrapyramidal motor effects (EPS), has greatly improved the tolerability of antipsychotic treatments. The pharmacology of atypical antipsychotics is varied and, although dopamine D2 receptor antagonism is common to all antipsychotics, the mechanisms of a typicality are complex and not fully understood. Thus, antagonism at 5-HT2 and/or other receptors, weak dopamine receptor affinity and, most recently, partial agonism at dopamine D2 receptors, have been variously implicated. However, because EPS have diminished with improvements in drug treatment, drug induced weight gain has emerged as a major concern, and the pharmacological basis of this problem, involving effects at 5-HT2c and perhaps other receptors, is yielding to investigation. Some drugs, notably the D2 partial agonists, can provide antipsychotic effects without the emergence of several of the seproblematic side-effects, which bodes well for future treatment. PMID- 15358978 TI - A positron emission tomography study of the 5-HT1A receptor in schizophrenia and during clozapine treatment. AB - Several post-mortem studies have identified increases of 5-HT1A receptor density in frontal cortical areas in schizophrenic patients, and one has found increases in the cerebellar vermis. Clozapine has moderate affinity at the 5-HT1A receptor, and this may be of therapeutic importance. This positron emission tomography (PET) study attempted to replicate the post-mortem findings in vivo and sought an occupancy effect of clozapine at the 5-HT1A receptor. We recruited healthy controls, and patients with schizophrenia who were divided into those receiving clozapine and those receiving neuroleptics lacking 5-HT1A receptor affinity. Each volunteer received a PET scan, using the 5-HT1A receptor radioligand [carbonyl 11C]WAY-100635, and a magnetic resonance imaging scan. The cerebellar vermis was examined by comparing time-activity data between groups. For other brain regions (the raphe and subdivisions of the cerebral cortex), binding potential images were generated to reflect receptor density, then analysed using 'region of interest' and voxel-by-voxel methods. No significant changes of 5-HT1A receptor density were found in schizophrenic patients compared to controls. Two other PET studies, containing drug naive rather than medicated schizophrenic patients, have also reported no increase in 5-HT1A receptor density in the frontal cortex. The results obtained in vivo bring into question the importance of the receptor in the pathophysiology of the illness. Clozapine did not occupy the 5-HT1A receptor at clinical doses. This is consistent with recent related PET results: 5-HT1A agonists do not appear to measurably block the binding of antagonist radiotracers in man at doses that are pharmacologically active but which are limited by tolerability. PMID- 15358979 TI - Cerebral D2 and 5-HT2 receptor occupancy in Schizophrenic patients treated with olanzapine or clozapine. AB - We report the results of a double-blind, randomized prospective trial on D2 and 5 HT2 receptor occupancy and the clinical effects of olanzapine versus clozapine in a sample of neuroleptic-refractory schizophrenic patients. Receptor occupancy was evaluated in different cortical areas and in basal ganglia using [18F] fluoro ethyl-spiperone ([18F] FESP) and positron emission tomography (PET). A total of 15 neuroleptic-free patients completed the study undergoing a baseline and a post treatment PET scan (olanzapine, nine patients, one female; clozapine, six patients, three female) 8 weeks after starting treatment. PET data were analysed both by regions of interest and on a voxel-by-voxel basis using Statistical Parametric Mapping (SPM96). Olanzapine and clozapine induced a similar and significant inhibition of [18F] FESP binding index in the cortex. In the basal ganglia, receptor occupancy was significantly higher with olanzapine than with clozapine (p=0.0018). By contrast, no differences in receptor occupancy were detected at the level of the pituitary gland. Clinical outcomes, in particular a full extra pyramidal tolerability, were similar. In this sample of neuroleptic refractory schizophrenic patients, olanzapine and clozapine showed a different pattern of occupancy of D2-like receptor despite a common lack of extrapyramidal side-effects. PMID- 15358980 TI - The relationship of sex hormones and cortisol with cognitive functioning in Schizophrenia. AB - Gonadal as well as stress hormones have recently been implicated in pathophysiology and sex differences in onset, prognosis and treatment of schizophrenia. The present study investigated the effects of serum levels of oestrogen, progesterone, testosterone and cortisol on neuropsychological functioning and psychopathology in a group of 37 patients (17 women, 20 men) with schizophrenia. Neuropsychological measures included tests of attention, verbal abilities, language, memory, executive functioning, motor and speed of information processing. The results showed that oestrogen and age was associated with low positive symptom scores, and within gender, cortisol predicted poor performance on the information processing domain in men. These findings demonstrate that cortisol, in addition to the commonly reported effects of oestrogen, influences neuropsychological functioning in schizophrenia with differential effects on specific domains of cognitive functioning and underscore the need for further investigation of the modulating role of hormones on neuropsychological functioning in schizophrenia. PMID- 15358981 TI - Mechanism of action of aripiprazole predicts clinical efficacy and a favourable side-effect profile. AB - The antipsychotic efficacy of aripiprazole is not generally associated with extrapyramidal symptoms, cardiovascular effects, sedation or elevations in serum prolactin that characterize typical or atypical antipsychotics. The aim of this study was to clarify the mechanism of action of aripiprazole that underlies its favourable clinical profiles. The preclinical efficacy and side-effect profiles of aripiprazole were evaluated using several pharmaco-behavioural test systems in mice and rats, both in vivo and ex vivo, and compared with those of other conventional and atypical antipsychotics. Each of the antipsychotics induced catalepsy and inhibited apomorphine-induced stereotypy. The catalepsy liability ratios for these drugs were 6.5 for aripiprazole, 4.7 for both olanzapine and risperidone. The ptosis liability ratios for aripiprazole, olanzapine and risperidone were 14, 7.2 and 3.3, respectively. Aripiprazole slightly increased DOPA accumulation in the forebrain of reserpinised mice, reduced 5-HTP accumulation at the highest dose and exhibited a weaker inhibition of 5-methoxy N,N-dimethyl-tryptamine-induced head twitches. Aripiprazole did not inhibit physostigmine- or norepinephrine-induced lethality in rats. In conclusion, aripiprazole shows a favourable preclinical efficacy and side-effect profile compared to a typical antipsychotics. This profile may result from its high affinity partial agonist activity at D2 and 5-HT1A receptors and its antagonism of 5-HT2A receptors. PMID- 15358982 TI - No association between akathisia or Parkinsonism and suicidality in treatment resistant Schizophrenia. AB - Akathisia and drug-induced Parkinsonism have traditionally been associated with depression and suicidality based on case study evidence. In this subanalysis, patients with treatment resistant schizophrenia were rated on the Comprehensive Psychopathological Rating Scale, Barnes Akathisia Scale and Simpson-Angus extrapyramidal side-effect scale at two time points (n=86 at first assessment; n=67 at second assessment). At no time point was there any significant relationship between akathisia and depression/suicidality or distress associated with akathisia and Parkinsonism with suicidality. These preliminary findings warrant further investigation. PMID- 15358983 TI - Increasing dopaminergic activity: effects of L-dopa and bromocriptine on human sensory gating. AB - Schizophrenic patients show a loss of sensory gating, which is reflected in a reduced P50 suppression. Because most of the symptoms in schizophrenia can be reduced by antagonists of the dopaminergic (D2) system, the loss in sensory gating might be related to an increased dopaminergic activity. Therefore, in the present study, the effects of increased dopaminergic neurotransmisson on sensory gating in healthy volunteers were investigated. In a double-blind, balanced, placebo-controlled design, healthy male volunteers were challenged in two separate studies with either 300 mg L-dopa (precursor of dopamine) or placebo (n=16) and 1.25 mg bromocriptine (D2 agonist) or placebo (n=17). Subsequently, they were tested for their sensory gating (P50 suppression). P50 suppression values in the placebo condition were comparable to those found in literature. Although both L-dopa and bromocriptine reduced P50 amplitude, they did so in an equal ratio for both the response to the conditioning (C) and the testing (T) stimuli, therefore not resulting in a reduction of the P50 suppression ratio (T/C). In the present study, neither L-dopa nor bromocriptine reduced sensory gating in healthy volunteers. This suggests that an increased dopaminergic activity in humans is not responsible for the reduction in sensory gating as seen, for example, in schizophrenia. PMID- 15358984 TI - The role of alpha1- and alpha2-adrenoreceptors on venlafaxine-induced elevation of extracellular serotonin, noradrenaline and dopamine levels in the rat prefrontal cortex and hippocampus. AB - The role of adrenergic alpha1- and alpha2-adrenoreceptors in augmentation of venlafaxine-induced elevation of extracellular serotonin (5-HT),noradrenaline (NA) and dopamine (DA) levels in the rat prefrontal cortex (PFC) and hippocampus (HIPP) was studied by in vivo microdialysis in anaesthetized rats. The alpha1 adrenoreceptor antagonist prazosin given alone (0.3 mg/kg, s.c.) induced only a moderate reduction of hippocampal 5-HT and NA levels. The alpha2-adrenoreceptor antagonist idazoxan (1.5 mg/kg, s.c.) causes moderate increases in the levels of 5-HT and DA in the PFC. The mixed 5-HT and NA reuptake inhibitor venlafaxine (10 mg/kg, i.p.) increased the efflux of 5-HT, NA and DA almost equally, to approximately 200% of the control levels in the PFC. The levels of 5-HT increased to 310%, an effect approximately twice the effect on NA in the HIPP. Venlafaxine also produced a moderate increase in DA levels in the PFC but had no effect in the HIPP. Pre-treatment with prazosin caused a significant attenuation of the venlafaxine induced 5-HT effect in the PFC, and a moderate increase in DA levels in the HIPP. Prazosin had no significant effect on the venlafaxine-induced increase of the NA levels in PFC or HIPP. A combined treatment of venlafaxine with idazoxan increased the venlafaxine NA and DA effects in PFC by a factor of two and resulted in a very robust five-fold augmentation of NA and DA concentrations in the HIPP. In summary, idazoxan was found to produce a potent enhancement of the venlafaxine effect to increase extracellular NA and DA levels in the PFC and, in particular, in the HIPP. Idazoxan had no effect on venlafaxine induced elevation of extracellular 5-HT levels in either PFC or HIPP and prazosin induced a decrease of 5-HT in the PFC. The present data suggest that blockade of alpha2-adrenoreceptors may play an important role in augmentation of the effects of mixed monoamine reuptake inhibitors. PMID- 15358985 TI - Antidepressant-like effect of tramadol and its enantiomers in reserpinized mice: comparative study with desipramine, fluvoxamine, venlafaxine and opiates. AB - Tramadol is a centrally acting analgesic that demonstrates opioid and monoaminergic properties. Several studies have suggested that tramadol could play a role in mood improvement. Moreover, it has previously been shown that tramadol is effective in the forced swimming test in mice and the learned helplessness model in rats, two behavioural models predictive of antidepressant activity. The aim of the present study was to test tramadol and its enantiomers in the reserpine test in mice, a classical observational test widely used in the screening of antidepressant drugs. This test is a non-behavioural method where only objective parameters such as rectal temperature and palprebral ptosis are considered. Moreover, we compared the effects of tramadol and its enantiomers with those of antidepressants (desipramine, fluvoxamine and venlafaxine) and opiates [morphine (-)-methadone and levorphanol]. Racemic tramadol, (-)-tramadol, desipramine and venlafaxine reversed the reserpine syndrome (rectal temperature and ptosis), whereas(+)-tramadol and fluvoxamine only antagonized the reserpine induced ptosis, without any effect on temperature. Opiates did not reverse reserpine-induced hypothermia. (-)-Methadone showed slight effects regarding reserpine-induced ptosis, morphine and levorphanol had no effect. These results show that tramadol has an effect comparable to clinically effective antidepressants in a test predictive of antidepressant activity, without behavioural implications. Together with other clinical and experimental data, this suggests that tramadol has an inherent antidepressant-like (mood improving) activity, and that this effect could have clinical repercussions on the affective component of pain. PMID- 15358986 TI - Effect of repeated ('binge') dosing of MDMA to rats housed at normal and high temperature on neurotoxic damage to cerebral 5-HT and dopamine neurones. AB - The technique of 'binge' dosing (several doses in one session) by recreational users of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) requires evaluation in terms of its consequences on the acute hyperthermic response and long-term neurotoxicity. We examined the neurotoxic effects of this dosing schedule on 5-HT and dopamine neurones in the rat brain. When repeated (three) doses of MDMA (2, 4 and 6 mg/kg i.p.) were given 3 h apart to rats housed at 19 degrees C, a dose dependent acute hyperthermia and long-term loss of 5-HT was observed in several brain regions (hippocampus, cortex and striatum), with an approximate 50% loss following 3 x 4 mg/kg and 65% decrease following 3 x 6 mg/kg. No decrease in striatal dopamine content was detected. When MDMA (4 mg/kg i.p.) was given repeatedly to rats housed at 30 degrees C, a larger acute hyperthermic response than that observed in rats treated at 19 degrees C environment was seen (maximum response 2.6 +/- 0.1 degrees C versus 1.3 +/- 0.2 degrees C). A long-term cerebral 5-HT loss of approximately 65% was also detected in both the cortex and hippocampus, but no loss in striatal dopamine content occurred. These data emphasize the increased acute hyperthermic response and neurotoxicity which occurs when MDMA is administered in a hot room environment compared to normal room temperature conditions, and support the view that MDMA is a selective 5-HT neurotoxin, even when a binge dosing schedule is employed and the rats are present in a hot environment. PMID- 15358987 TI - Effects of reboxetine on sleep and nocturnal cardiac autonomic activity in patients with dysthymia. AB - Antidepressants may have sleep and autonomic side-effects. The acute and long term effect of reboxetine (2 mg b.i.d.) on sleep and cardiac autonomic activity was compared with that of placebo in a single-blind study. Twelve patients affected by dysthymia underwent four polysomnographic studies at baseline (placebo); at night 3 (reboxetine; acute effect); at night 9 (reboxetine; intermediate-term effect); and at night 122 (reboxetine; chronic effect). After the first administration, reboxetine increased time awake after sleep onset, number of awakenings, percentage of stages 1 and 2 non-rapid eye movement (REM), and reduced the amount of stages 3-4 non-REM, but all these effects disappeared by continuing treatment. However, reboxetine caused a persistent suppression of REM sleep, which was accompanied by an increase of REM sleep latency. The spectral analysis of heart rate variability showed a trend towards an increase in sympathetic activity with both acute and intermediate reboxetine use. Long-term treatment with 4 mg reboxetine does not cause significant changes in cardiac autonomic function. PMID- 15358988 TI - Use of reboxetine in bulimia nervosa: a pilot study. AB - The pharmacological approach to bulimia nervosa is mainly based (BN) on selective serotonin reuptake inhibitors, but many elements suggest the possible involvement of the noradrenergic system in this disorder. The aim of the study was to assess the efficacy of reboxetine--a selective norepinephrine uptake inhibitor--in a sample of bulimic outpatients, after 3 months of treatment. Twenty-eight of 77 consecutively admitted patients with a DSM-IV diagnosis of BN (without Axis I comorbidity) received reboxetine. All patients were assessed at baseline (T0), and after 1 month (T1) and 3 months (T3), respectively, of treatment with reboxetine 4 mg/day. The subjects were administered the following questionnaires: Hamilton Rating Scale for Anxiety (HAM-A) and for Depression (HAM-D), Global Assessment Functioning (GAF), Eating Disorder Inventory-2 (EDI-2) and Body Shape Questionnaire (BSQ). Sixty percent of the patients were responsive to treatment(evaluated as a 50% decrease of bulimic behaviours). After 3 months of treatment, a significant reduction emerged in the scores of various EDI-2 subscales (Bulimia, Drive for Thinness, Body Dissatisfaction, Social Insecurity, Interpersonal Distrust, etc.) and in the BSQ total score. Moreover, depressive symptoms (HAM-D) and Global Functioning (GAF) scores showed a significant improvement. These data support a fast and favourable effect of reboxetine in the treatment of BN, both on symptoms and psychopathological features. Moreover, the specific and strong action of reboxetine on improvement of social functioning is also supported in this disorder. PMID- 15358989 TI - The effects of clozapine and high-dose olanzapine on brain function in treatment resistant schizophrenia: a case study. AB - Although demonstrating superior efficacy in people with treatment-resistant schizophrenia, clozapine may cause serious side effects, requires blood monitoring and is costly to administer. Olanzapine is similar to clozapine in molecular structure and pharmacologic action but has not demonstrated as robust results as clozapine at routine doses (10-25 mg). Here we present a case study measuring blood flow by positron emission tomography (PET) imaging for a patient treated sequentially with a high dose of olanzapine (50 mg/day) followed by clozapine each for 8 weeks in a double-blind design. During a task, clozapine produced more brain activation patterns than during treatment with olanzapine or during the drug free condition (2 week washout). Clozapine resulted in recruitment of frontal, parietal and cingulate regions that did not appear to be active during olanzapine in this 44 year old right handed male. Additionally, a more robust decrease in symptoms was noted on the Brief Psychiatric Rating Scale (BPRS) score than with olanzapine treatment. These findings suggest that high doses of olanzapine do not produce similar brain activation patterns as clozapine in people with treatment-resistant schizophrenia. PMID- 15358990 TI - Rhabdomyolysis and compartment syndrome with coadministration of risperidone and simvastatin. AB - We report a case of rhabdomyolysis and acute compartment syndrome of the lower extremity in a schizophrenic patient taking risperidone following the addition of simvastatin to treat hyperlipidemia. We suspect that disrupted drug metabolism, resulting from interactions with cytochrome P450 enzymes, rapidly elevated drug plasma levels, which then led to muscle toxicity. Clinicians who pharmacologically treat medical comorbidities in patients receiving atypical antipsychotics must be proactive in anticipating potential drug-drug interactions. PMID- 15358991 TI - Diabetic keto-acidotic (DKA) coma following olanzapine initiation in a previously euglycaemic woman and successful continued therapy with olanzapine. AB - We report the case of a euglycaemic woman whose glucose control rapidly decompensated following olanzapine initiation leading to diabetic coma. Hyperglycaemia has been associated with chronic psychotic disorders and antipsychotics for many years. However, it is unusual to see such rapid and life threatening changes associated with treatment. The case highlights that changes in antipsychotic treatment may be associated with large changes in glucose tolerance, and that it is possible to continue antipsychotic treatment with appropriate diabetic care. PMID- 15358992 TI - Magnesium homeostasis and antipsychotic-induced QTc prolongation. PMID- 15358993 TI - Vigabatrin and tiagabine might have antipanic properties. PMID- 15358994 TI - Changes in pharmacological treatment for bipolar disorder over time in Manchester: a comparison with Lloyd et al. (2003). PMID- 15358995 TI - Reorganizing the system of care surrounding laparoscopic surgery: a cost effectiveness analysis using discrete-event simulation. AB - PURPOSE: To determine the cost-effectiveness of a proposed reorganization of surgical and anesthesia care to balance patient volume and safety. METHODS: Discrete-event simulation methods were used to compare current surgical practice with a new modular system in which patient care is handed off between 2 anesthesiologists. A health care system's perspective, using hospital and professional costs, was chosen for the cost-effectiveness analysis. Outcomes were patient throughput, flow time, wait time, and resource use. Sensitivity analyses were performed on staffing levels, mortality rates, process times, and scheduled patient volume. RESULTS: The new strategy was more effective (average 4.41 patients/d [median = 5] v. 4.29 [median = 4]) and had similar costs (average cost/ patient/d = 5327 dollars v. 5289 dollars) to the current strategy with an incremental cost-effectiveness of 318 dollars/additional patient treated/d. Surgical mortality rate must be >4% or hand-off delay >15 min before the new strategy is no longer more effective. CONCLUSION: The proposed system is more cost-effective relative to current practice over a wide range of mortality rates, hand-off times, and scheduled patient volumes. PMID- 15358996 TI - Pramipexole v. levodopa as initial treatment for Parkinson's disease: a randomized clinical-economic trial. AB - PURPOSE: To determine the 2-year incremental cost effectiveness of initial pramipexole treatment compared with initial levodopa treatment in patients with early Parkinson's disease (PD). METHODS: 301 subjects with early PD were randomized to either pramipexole or levodopa and followed every 3 months over a 2 year period. Costs were assigned to patient collected health utilization data using a variety of methods. Health state preferences were estimated using the EuroQol. RESULTS: Pramipexole strategy was an estimated 2,138 dollars (SE = 1,182 dollars) more expensive than levodopa strategy. The incremental cost effectiveness of pramipexole compared with levodopa was 106,900 dollars/QALY (EQ 5D), compared with pramipexole being dominated by levodopa using the EQVAS. CONCLUSIONS: Although considerable uncertainty exists in the 2-year cost effectiveness of initial pramipexole compared with initial levodopa in the treatment of early PD, our estimates suggest that pramipexole may not be welfare enhancing during the first 2 years of treatment. If initial pramipexole results in long-term improvements in quality of life, its cost-effectiveness will become more favorable. PMID- 15358997 TI - The cost-effectiveness of screening programs using single and multiple birth cohort simulations: a comparison using a model of cervical cancer. AB - Despite early recognition of the theoretical advantages of simulations that include different population subgroups/ strata and different birth cohorts, many modeling-based economic evaluations of cervical screening have been based on unrealistic single birth cohort simulations. The authors examined the effect of a multiple birth cohort simulation on the incremental cost-effectiveness estimates of cervical screening programs, compared to a conventional single cohort simulation. The choice of hypothetical cohort that starts the simulation had a major impact on the cost-effectiveness estimates: Compared with a single birth cohort simulation, the incremental cost-effectiveness of a shift from biennial to triennial screening was 30% higher when using the multiple cohort simulation. Multiple cohort simulations using the different age structures of 4 countries had little impact on the cost effectiveness ratios (variation <5%). Future modeling based evaluations of screening policies should better reflect the age range of the population that is targeted by carefully specifying the nature of the starting cohort(s). PMID- 15358998 TI - A comparison of different strategies to collect standard gamble utilities. AB - OBJECTIVE: The authors performed a methodological comparison of the usual standard gamble with methods that could also be used in mailed questionnaires. METHODS: Ninety-two diabetic patients valued diabetes-related health states twice. In face-to-face interviews, the authors used an iterative standard gamble (ISG) in which the probabilities were varied in a ping-pong manner and a self completion method (SC) with top-down titration as search procedure (SC-TD) in 2 independent subsamples of 46 patients. Three months later, all patients received a mailed questionnaire in which the authors used the self-completion method with bottom-up (SCBU) and SC-TD as search procedures. RESULTS: ISG and SCTD showed feasibility and consistency in the interviews. The ISG resulted in significantly higher utilities than the SC-TD. Two thirds of the mailed questionnaires provided useful results indicating some problems of feasibility. Utilities measured by SC BU and SC-TD did not differ significantly showing procedural invariance. Further, patients indicated ambivalence when given the choice not to definitely state their preferences. CONCLUSIONS: The results show that different strategies to collect standard gamble utilities can yield different results. Compared with the usually applied ISG, the SC method is feasible in interviews and provides a consistent alternative that is less costly when used in mailed questionnaires, although its practicability has to be improved in this later setting. PMID- 15358999 TI - Preference assessment method affects decision-analytic recommendations: a prostate cancer treatment example. AB - PURPOSE: To evaluate the effect of preference assessment method on treatment recommended by an individualized decision-analytic model for early prostate cancer. METHODS: Health state preferences were elicited by time tradeoff, rating scale, and a power transformation of the rating scale from 63 men ages 55 to 75. The authors used these values in a Markov model to determine whether radical prostatectomy or watchful waiting yielded the greater quality-adjusted life expectancy. RESULTS: Time tradeoff and transformed rating scale recommendations differed widely. Time tradeoff and transformed rating scale utilities differed in their treatment recommendation for 21% to 52% of men, and the mean difference in quality-adjusted life years varied from less than 0.5 to greater than 1.0. CONCLUSIONS: Treatment recommendations from the prostate cancer decision model were sensitive to the method of preference assessment. If decision analysis is used to counsel individual patients, careful consideration must be given to the method of preference elicitation. PMID- 15359000 TI - Correcting biases in standard gamble and time tradeoff utilities. AB - The standard gamble (SG) method and the time tradeoff (TTO) method are commonly used to measure utilities. However, they are distorted by biases due to loss aversion, scale compatibility, utility curvature for life duration, and probability weighting. This article applies corrections for these biases and provides new data on these biases and their corrections. The SG and TTO utilities of 6 rheumatoid arthritis health states were assessed for 45 healthy respondents. Various corrections of utilities were considered. The uncorrected TTO scores and the corrected (for utility curvature) TTO scores provided similar results. This article provides arguments suggesting that the TTO scores are biased upward rather than having balanced biases. The only downward bias in TTO scores was small and probably cannot offset the upward biases. TheTTOscores are higher than the theoretically most preferred correction of the SG, the mixed correction. These findings suggest that uncorrected SG scores, which are higher than TTO scores, are too high. PMID- 15359001 TI - Medical and nonmedical determinants of decision making about potentially life prolonging interventions. AB - Patient characteristics may influence medical decision making in various ways. The contribution of several patient characteristics to medical decision making was studied. Thirty oncologists, 29 nursing home physicians, and 22 cardiologists were interviewed (overall response = 60%). Respondents were asked whether they would apply a specified intervention for a number of hypothetical seriously ill patients, who varied with respect to factors that were not relevant to the outcome of treatment. The condition that made patients clearly eligible for treatment was kept constant. In a multivariate regression model, patients with a better physical condition, a more obvious social role, and a lower age were more likely to be treated than were other patients. Medical decision making is not exclusively based on empirical evidence but also related to morally complex issues such as patient age and social status. PMID- 15359002 TI - Group sequential design for comparative diagnostic accuracy studies: implications and guidelines for practitioners. AB - PURPOSE: Comparative diagnostic accuracy (CDA) studies are typically small retrospective studies supporting a higher accuracy for one modality over another for either staging a particular disease or assessing response to therapy, and they are used to generate hypotheses for larger prospective trials. The purpose of this article is to introduce the group sequential design (GSD) approach in planning these larger trials. METHODS: Methodology needed for using GSD in the CDA studies is recently developed. In this article, GSD with the O'Brien and Fleming (OBF) stopping rule is described and guidelines for sample size calculation are provided. Simulated data is used to demonstrate the application of GSD in the design/analysis of a clinical trial in the CDA study setting. RESULTS: The expected sample size needed for planning a trial with GSD (under the OBF stopping rule) is slightly inflated but may ultimately result in greater savings of patient resources. CONCLUSION: GSD is a specialized statistical method that is helpful in balancing the ethical and financial advantages of stopping a study early against the risk of an incorrect conclusion and should be adopted for planning CDA studies. PMID- 15359003 TI - To err is human. PMID- 15359004 TI - Unfinished symphony: a tribute to the life and career of Bernie O'Brien (1959 2004). AB - The death of Bernie O'Brien in February 2004 brought a premature end to one of the most productive and influential careers in the area of health technology assessment and economic evaluation. A long-term member of the Society for Medical Decision Making, Bernie will be remembered for his research contributions in areas including outcome valuation, decision modeling, statistical methods in economic evaluation, and applied cost-effectiveness studies. He was also an excellent communicator and teacher and, above all, a fun guy to work with. In this article, the authors provide a review of Bernie's academic contributions. PMID- 15359007 TI - Integration of retrograde axonal and nuclear transport mechanisms in neurons: implications for therapeutics. AB - The elongated morphology of neuronal processes imposes a significant challenge for effective intracellular communication between the neurites and the cell body. This problem is especially acute upon injury, when the cell body must receive accurate and timely information on the site and extent of axonal damage to mount an appropriate response. Recent work has demonstrated that nuclear import factors from the importin (karyopherin) alpha and beta families provide a mechanism for retrograde injury signaling. Importins are found throughout axons and dendrites at significant distances from the cell body, and importin beta protein is increased after nerve lesion by local translation of axonal mRNA. This leads to formation of a high-affinity nuclear localization signal (NLS) binding complex that traffics retrogradely due to an interaction of importin alpha with the motor protein dynein. Disruption of the complex with excess NLS peptides delays regeneration of injured sensory neurons. The dual role of importins in retrograde transport in axons and nuclear import in cell bodies suggests new avenues for manipulating intrinsic regeneration mechanisms in the nervous system and may provide a novel route for drug delivery to the CNS. PMID- 15359008 TI - Retinoic acid signaling in the nervous system of adult vertebrates. AB - The majority of the functions of vitamin A are carried out by its metabolite, retinoic acid (RA), a potent transcriptional activator acting through members of the nuclear receptor family of transcription factors. In the CNS, RA was first recognized to be essential for the control of patterning and differentiation in the developing embryo. It has recently come to light, however, that many of the same functions that RA directs in the embryo are involved in the regulation of plasticity and regeneration in the adult brain. The same intricate metabolic control system of synthetic and catabolic enzymes, combined with cytoplasmic binding proteins, is used in both embryo and adult to create regions of high and low RA to modulate gene transcription. This review summarizes some of the discoveries in the new field of retinoid neurobiology including its functions in neural plasticity and LTP in the hippocampus; its possible role in motor disorders such as Parkinson's disease, motoneuron disease, and Huntington's disease; its role in regeneration after sciatic nerve and spinal cord injury; and its possible involvement in psychiatric diseases such as depression. PMID- 15359009 TI - Brain angiotensin II and synaptic transmission. AB - The renin-angiotensin system is an enzymatic cascade by which angiotensinogen is cleaved by renin and then by angiotensin-converting enzyme to produce angiotensin II (Ang II) and subsequently other angiotensins. Biochemical and neurophysiological studies have documented the presence of the reninangiotensin system and specific Ang II receptors in the brain. Also, circulating Ang II can exert some of its actions, such as blood pressure control and body fluid homeostasis, through stimulation of Ang II receptors in the circumventricular organs that lack a normal blood-brain barrier. In addition to some of the post synaptic effects of Ang II, recent studies have revealed that Ang II regulates synaptic transmission in several brain regions, especially the nucleus of the solitary tract, hypothalamic paraventricular nucleus, and hippocampus. This review summarizes emerging new evidence on the effect of brain Ang II on glutamatergic and GABAergic synaptic transmission. This previously unrecognized presynaptic action of Ang II is important for the control of neuronal excitability and many physiological functions including autonomic control, hormone secretion, and memory. Future research on the role of brain-derived Ang II and its receptors in synaptic transmission will further enhance our understanding of the cellular mechanisms of Ang II and the relationship between the renin-angiotensin system and brain functions. PMID- 15359010 TI - Zn2+ ions: modulators of excitatory and inhibitory synaptic activity. AB - The role of Zn(2+) in the CNS has remained enigmatic for several decades. This divalent cation is accumulated by specific neurons into synaptic vesicles and can be released by stimulation in a Ca(2+)-dependent manner. Using Zn(2+) fluorophores, radiolabeled Zn(2+), and selective chelators, the location of this ion and its release pattern have been established across the brain. Given the distribution and possible release under physiological conditions, Zn(2+) has the potential to act as a modulator of both excitatory and inhibitory neurotransmission. Excitatory N-methyl-D-aspartate (NMDA) receptors are directly inhibited by Zn(2+), whereas non-NMDA receptors appear relatively unaffected. In contrast, inhibitory transmission mediated via GABA(A)receptors can be potentiated via a presynaptic mechanism, influencing transmitter release; however, although some tonic GABAergic inhibition may be suppressed by Zn(2+), most synaptic GABA receptors are unlikely to be modulated directly by this cation. In the spinal cord, glycinergic transmission may also be affected by Zn(2+) causing potentiation. Recently, the penetration of synaptically released Zn(2+) into neurons suggests that this ion has the potential to act as a direct transmitter, by affecting postsynaptic signaling pathways. Taken overall, present studies are broadly supportive of a neuromodulatory role for Zn(2+) at specific excitatory and inhibitory synapses. PMID- 15359011 TI - Neuronal survival and p73/p63/p53: a family affair. AB - Elucidating the mechanisms that regulate the life versus death of mammalian neurons is important not only for our understanding of the normal biology of the nervous system but also for our efforts to devise approaches to maintain neuronal survival in the face of traumatic injury or neurodegenerative disorders. Here, we review the emerging evidence that a key survival/death checkpoint in both peripheral and central neurons involves the p53 tumor suppressor and its newly discovered family members, p73 and p63. The full-length isoforms of these proteins function as proapoptotic proteins, whereas naturally occurring N terminal truncated variants of p73 and p63 act as prosurvival proteins, at least partially by antagonizing the full-length family members. The authors propose that together, these isoforms comprise an upstream rheostat that sums different environmental cues to ultimately determine neuronal survival during development, during neuronal maintenance in adult animals, and even following traumatic injury. PMID- 15359012 TI - Huntingtin and its role in neuronal degeneration. AB - Huntington's disease results from a polyglutamine expansion in the N-terminal region of huntingtin (htt). This abnormality causes protein aggregation and leads to neurotoxicity. Despite its widespread expression in the brain and body, mutant htt causes selective neurodegeneration in Huntington's disease patient brains. However, Huntington's disease mouse models expressing mutant htt do not have obvious neurodegeneration despite significant neurological symptoms. Most Huntington's disease mouse models display the accumulation of toxic N-terminal mutant htt fragments in both the nucleus and neuronal processes, suggesting that these subcellular sites are hotspots for the early neuropathology of Huntington's disease. Intranuclear htt affects gene expression and may cause neuronal dysfunction. Mutant htt in neuronal processes affects axonal transport and induces degeneration, and these effects may be more relevant to the selective neurodegeneration in Huntington's disease. Growing evidence has also suggested that mutant htt mediates multiple pathological pathways. This review discusses the early pathological changes identified in Huntington's disease cellular and animal models. These changes may be the causes of neurode-generation. PMID- 15359013 TI - Quantitative analysis of connectivity in the visual cortex: extracting function from structure. AB - It is generally agreed that information flow through the cortex is constrained by a hierarchical architecture. Lack of precise data on areal connectivity leads to indeterminacy of existing models. The authors introduce two quantitative parameters (SLN and FLN) that hold the promise of resolving such indeterminacy. In the visual system, using a very incomplete database, provisional hierarchies are in line with the recent proposal of higher functions of area V1 and suggest a hitherto unsuspected central function of the frontal eye field. PMID- 15359014 TI - Getting a cue: the need to belong and enhanced sensitivity to social cues. AB - To successfully establish and maintain social relationships, individuals need to be sensitive to the thoughts and feelings of others. In the current studies, the authors predicted that individuals who are especially concerned with social connectedness--individuals high in the need to belong--would be particularly attentive to and accurate in decoding social cues. In Study 1, individual differences in the need to belong were found to be positively related to accuracy in identifying vocal tone and facial emotion. Study 2 examined attention to vocal tone and accuracy in a more complex social sensitivity task (an empathic accuracy task). Replicating the results of Study 1, need to belong scores predicted both attention to vocal tone and empathic accuracy. Study 3 provided evidence that the enhanced performance shown by those high in the need to belong is specific to social perception skills rather than to cognitive problem solving more generally. PMID- 15359015 TI - Explaining the discrepancy between intentions and actions: the case of hypothetical bias in contingent valuation. AB - An experiment was designed to account for intention-behavior discrepancies by applying the theory of planned behavior to contingent valuation. College students (N = 160) voted in hypothetical and real payment referenda to contribute $8 to a scholarship fund. Overestimates of willingness to pay in the hypothetical referendum could not be attributed to moderately favorable latent dispositions. Instead, this hypothetical bias was explained by activation of more favorable beliefs and attitudes in the context of a hypothetical rather than a real referendum. A corrective entreaty was found to eliminate this bias by bringing beliefs, attitudes, and intentions in line with those in the real payment situation. As a result, the theory of planned behavior produced more accurate prediction of real payment when participants were exposed to the corrective entreaty. PMID- 15359016 TI - The correspondence between attachment to parents and God: three experiments using subliminal separation cues. AB - Attachment theoretical studies have increased our understanding of the socioemotional foundations for religious development. However, because these studies have been correlational and based on self-reports, they are vulnerable to concerns of self-presentation bias and lack of basis for causal inference. Three subliminal stimulation experiments were therefore performed, where activation of the attachment system was attempted by way of unconsciously administered separation stimuli. In Experiments 1 and 3 (N = 29 and 89), the separation stimulus alluded to God, and in Experiment 2 (N = 47), it alluded to mother. Responses were moderated by perceived attachment history with parents in all experiments. Participants with secure histories increased in religious attachment behaviors, whereas those with insecure histories decreased following attachment system activation compared with control stimulation. There also were suggestions of experimental group increase in proximity seeking in relation to God. The main conclusion supports correspondence between internal working models of parents and God. PMID- 15359017 TI - Deliver us from evil: the effects of mortality salience and reminders of 9/11 on support for President George W. Bush. AB - According to terror management theory, heightened concerns about mortality should intensify the appeal of charismatic leaders. To assess this idea, we investigated how thoughts about death and the 9/11 terrorist attacks influence Americans' attitudes toward current U.S. President George W. Bush. Study 1 found that reminding people of their own mortality (mortality salience) increased support for Bush and his counterterrorism policies. Study 2 demonstrated that subliminal exposure to 9/11-related stimuli brought death-related thoughts closer to consciousness. Study 3 showed that reminders of both mortality and 9/11 increased support for Bush. In Study 4, mortality salience led participants to become more favorable toward Bush and voting for him in the upcoming election but less favorable toward Presidential candidate John Kerry and voting for him. Discussion focused on the role of terror management processes in allegiance to charismatic leaders and political decision making. PMID- 15359018 TI - Effects of attitude action identification on congruence between attitudes and behavioral intentions toward social groups. AB - Attitude Representation Theory (ART) holds that attitude-relevant responses are informed by mental representations of the attitude object, which include the individual's actions toward that object. Action Identification Theory (AIT) holds that the same action can be identified at multiple levels. Individuals who identify their actions at lower levels have less flexibility in how they perform the action, and thus enact the action less consistently. An integration of ART and AIT suggested that individuals who spontaneously (Experiment 1) or through manipulation (Experiments 2 and 3) identify their attitude-relevant actions toward a social group at lower levels might display less attitude-intention congruence than would individuals who identify their attitude-relevant actions at higher levels. ART and AIT are discussed as having links with each other and with other theories of attitude and judgment processes. PMID- 15359019 TI - Religion and emotional compensation: results from a prospective study of widowhood. AB - Based on recent applications of attachment theory to religion, the authors predicted that the loss of a spouse would cause widowed individuals to increase the importance of their religious/spiritual beliefs. This hypothesis was examined using the Changing Lives of Older Couples sample from which preloss measures of religiosity were available for widowed individuals and matched controls. A total of 103 widowed individuals provided follow-up data, including reports of religious beliefs and grief, at 6 months, 24 months, and 48 months after the loss. Results indicated that widowed individuals were more likely than controls to increase their religious/spiritual beliefs. This increase was associated with decreased grief but did not influence other indicators of adjustment such as depression. Finally, insecure individuals were most likely to benefit from increasing the importance of their beliefs. Results are discussed in terms of the potential value of applying psychological theory to the study of religion. PMID- 15359020 TI - The name of the game: predictive power of reputations versus situational labels in determining prisoner's dilemma game moves. AB - Two experiments, one conducted with American college students and one with Israeli pilots and their instructors, explored the predictive power of reputation based assessments versus the stated "name of the game" (Wall Street Game vs. Community Game) in determining players' responses in an N-move Prisoner's Dilemma. The results of these studies showed that the relevant labeling manipulations exerted far greater impact on the players' choice to cooperate versus defect--both in the first round and overall--than anticipated by the individuals who had predicted their behavior. Reputation-based prediction, by contrast, failed to discriminate cooperators from defectors. A supplementary questionnaire study showed the generality of the relevant short-coming in naive psychology. The implications of these findings, and the potential contribution of the present methodology to the classic pedagogical strategy of the demonstration experiment, are discussed. PMID- 15359021 TI - Playboy Playmate curves: changes in facial and body feature preferences across social and economic conditions. AB - Past research has investigated ideals of beauty and how these ideals have changed across time. In the current study, facial and body characteristics of Playboy Playmates of the Year from 1960-2000 were identified and investigated to explore their relationships with U.S. social and economic factors. Playmate of the Year age, body feature measures, and facial feature measurements were correlated with a general measure of social and economic hard times. Consistent with Environmental Security Hypothesis predictions, when social and economic conditions were difficult, older, heavier, taller Playboy Playmates of the Year with larger waists, smaller eyes, larger waist-to-hip ratios, smaller bust-to waist ratios, and smaller body mass index values were selected. These results suggest that environmental security may influence perceptions and preferences for women with certain body and facial features. PMID- 15359022 TI - Organization of partner knowledge: relationship outcomes and longitudinal change. AB - This study examined the association between organization of knowledge about a romantic partner (partner structure) and relationship status (ongoing or ended) 1 year later. Ironically, partner structures that were associated with more positive feelings about one's partner at Time 1 were associated with greater rates of breakup by Time 2. These results are interpreted in terms of the vulnerability of compartmentalized partner structures to shifts in the salience of negative beliefs and the hypothesized difficulty of maintaining integrative structures for an extended period of time. Change in partner structure during 1 year's time was consistent with the predictions of the dynamic model that evaluative integration should increase when negative beliefs become salient. Such change (which may represent a transient shift) was associated with couples' longevity when relationship conflict was low, supporting the view that integration reflects a struggle with negative attributes that may or may not be successful. PMID- 15359023 TI - Levels of interpersonal complementarity: a simplex representation. AB - The relations among different measures of interpersonal behavior and complementarity across level were examined in one session of a sample of therapy dyads (N = 26) and in an interaction between college students (N = 108). Four levels of complementarity, trait, aggregate situation, behavioral interchanges, and behavioral interchanges with base rates removed were examined as they covaried among themselves and with interaction evaluations. The four levels of complementarity were found to be fit by a simplex structure, and this structure was related to interaction evaluation in both samples. The complementarity evaluation relation was mediated by base-rate-corrected complementarity. Implications relative to the operationalization of complementarity are discussed. PMID- 15359024 TI - Somatic and social: Chinese Americans talk about emotion. AB - Empirical findings suggest that Chinese and Americans differ in the ways that they describe emotional experience, with Chinese using more somatic and social words than Americans. No one, however, has investigated whether this variation is related to differences between Chinese and American conceptions of emotion or to linguistic differences between the English and Chinese languages. Therefore, in two studies, the authors compared the word use of individuals who varied in their orientation to Chinese and American cultures (European Americans [EA], more acculturated Chinese Americans [CA], and less acculturated CA) when they were speaking English during emotional events. Across both studies, less acculturated CA used more somatic (e.g., dizzy) and more social (e.g., friend) words than EA. These findings suggest that even when controlling for language spoken, cultural conceptions of emotion may shape how people talk about emotion. PMID- 15359025 TI - The many meanings of unitary: a plea for clarity. PMID- 15359026 TI - The concept of branding: is it relevant to nursing? AB - This concept exploration examines branding and its relevance to nursing. Branding is used to differentiate products through use of symbols. The symbols are the brands that are designed to communicate the value of products. Nursing has had many identifying symbols, such as the nurse's cap and the white uniform, but these symbols have failed to clearly communicate the essence of nursing. Lack of a distinct nursing brand has led to confusion about the discipline. The Roy adaptation model provides a view of branding as a process for clearly defining the profession, improving its image, and differentiating its role within the healthcare milieu. PMID- 15359027 TI - A dialogue about two nursing science traditions: the Roy adaptation model and the human becoming theory. AB - The authors engage in a dialogue about the ontology, epistemology, and methodologies in research within their respective theoretical perspectives, the Roy adaptation model and the human becoming theory, insofar as these represent different paradigms in nursing science. In the discussion of ontology as the lens through which humans view their world, they articulate the primacy of ontology over methodology in shaping knowledge development. PMID- 15359028 TI - Ethics content in nursing education: pondering with the possible. AB - Accrediting bodies of nursing as well as other healthcare organizations require that nursing ethics content be incorporated in programs of study. How does nursing define the termnursing ethics? How is it distinguished or viewed in various levels of higher degree nursing education? What is the role of codes for nursing? These are but a few of the questions pondered in the following article that includes personal correspondence with two distinguished authors, educators, and consultants for biomedical ethics, Dr. Sara T. Fry and Dr. Nancy Berlinger of the Hastings Center. PMID- 15359030 TI - Does experience really matter? PMID- 15359031 TI - Mentoring: a human becoming perspective. AB - There have been reports of an absence or a lack of support for one another within the discipline of nursing. By establishing patterns of engagement, seasoned veterans connect with their novice colleagues to promote standards of excellence and scholarship. In order to provide the multidimensional benefits of a mentorship opportunity, a clear theoretical vision of mentoring is required. The theoretical perspective of human becoming provides a framework for mentors to establish gentle urgings with their mentees and thus provide for the growing of nurses as well as the discipline of nursing. PMID- 15359033 TI - Human becoming: scope and challenges continued. Interview by Jacqueline Fawcett. PMID- 15359034 TI - Musical expressions of life: a look at the 18th and 19th century from a human becoming perspective. AB - What follows is an exploration of 18th and 19th century music of the Western world through a nursing science lens, specifically that of the human becoming theory. This article was written while I was enrolled in a music history course, which afforded me the opportunity to explore music as musical expressions of life. Rooted in the human becoming philosophical perspective, which focuses on unitary human experience and the quality of human life, I discuss musical expressions of life with examples from various composers throughout the 18th and 19th century. This article concludes with a reflection on musical expressions and their contribution to the enhancement of the quality of human life, a focus of nursing from a human becoming perspective. PMID- 15359035 TI - Life-closing spirituality and the philosophic assumptions of the Roy adaptation model. AB - Secondary analysis of data from a previous study that referenced spirituality was coded, categorized, and grouped into themes. Life-closing spirituality for 44 (45.4%) of 97 total participants was shaped by a core theme of believing that was central to dying persons. Believing was linked to six other themes: comforting, releasing, connecting, giving, reframing, and requesting. These themes supported the philosophic assumptions and principles of humanism and veritivity as defined in the Roy adaptation model. PMID- 15359036 TI - Similarities and differences in experiences of hope. AB - From a human becoming perspective, hope is considered to be a universal lived experience, which arises and fades in the human-universe process. This article is a comparison of six Parse research method studies on the lived experience of hope from five countries: Japan, Taiwan, Sweden, Finland, and the United States. The goal was to explore the similarities and differences in the findings of a nine country study on hope. While the structure of the experience of hope was similar, some differences are discussed that enhance nursing practice and guide further study. PMID- 15359037 TI - Theory of dependent-care in research with parents of toddlers: the NEAT project. AB - This paper contains a description of the use of the theory of dependent-care in the Nutrition Aimed at Toddlers project, a nursing intervention study with parents of toddlers, and discusses the research and practice utility of the theory of dependent-care, a corollary theory to Orem's self-care theory. In particular, the concepts of dependent-care, dependent-care agency, therapeutic self-care demand and an educative-supportive nursing system are discussed within the context of the Nutrition Aimed at Toddlers project. PMID- 15359040 TI - Using the right tool for the job. PMID- 15359039 TI - Current challenges and issues facing nursing in Australia. AB - This column is a discussion of nursing in Australia. The authors present current challenges and issues related to recruitment and retention, an aging workforce, violence and bullying, diversity, undergraduate and graduate education, professional development, information technology, clinical pathways, models of care delivery, and funding models. They also present what is happening in Australia in relation to nursing's scope of practice, development of the nurse practitioner role, nursing leadership, and nursing research. PMID- 15359041 TI - Doctors, "dirty work" patients, and "revolving doors". AB - Using information from research into the phenomenon of "revolving-door" psychiatric patients, the author explores general practitioners' perceptions of difficult patients and the consequences for patient management. He first considers the concept of "good and bad" as a possible subtype of dirty work. He then presents the evidence of medical irritation with patients from interview data and explores the rationalizations for the way in which patients are subsequently managed. In line with previous studies, the author argues that the construction of patients as difficult and the subsequent dynamics of exclusion lie in the breakdown of the "normal" doctor-patient relationship coupled with the doctor's need to get on with the day's workload. Moral judgments formed a part of the exclusion process. PMID- 15359042 TI - Being present in a distant room: aspects of teleconsultations with older people in a nursing home. AB - In a telecare project in Northern Sweden, videophones have been used to facilitate teleconsultations between nurses and elders at a nursing home. The authors designed this study to elucidate qualities in the communication in the professional encounter between nurses and elders assisted by nursing staff in the teleconsultations. They interviewed 2 registered nurses and 5 nursing staff members with long experience of using videophones in the telecare project, out of 20 staff members, and analyzed them using a phenomenological-hermeneutic method. In a comprehensive interpretation, teleconsultations could be understood as glimpses of the experience of being in the other's room with a feeling of providing nursing presence. This was attained when aspects such as familiarity, safety, transparency, and interest were promoted. PMID- 15359043 TI - Strategies used by low-income Mexican women to deal with miscarriage and "spontaneous" abortion. AB - This study focuses on lowest income Mexican women attended for abortion-related complications in a public hospital. The objective was to investigate the women's experience of having a so-called "spontaneous" abortion and their related strategies to avoid stigmatization. Four strategies emerge from women's testimonies: presenting themselves as women who "play by the rules," pleading ignorance of the pregnancy, stating that they had already accepted their pregnancy, or presenting the abortion as the result of an accident. Women use these strategies to deflect any blame to which they might be subjected and as a means of dealing with the stigma attached to a behavior that transgresses social norms regarding reproduction. Far from being passive receptors of the social imperative, which makes motherhood compulsory, women oscillate strategically within the margins of a seemingly uniform normative discourse and thereby ensure their moral survival. The authors discuss results within the framework of praxis theory. PMID- 15359044 TI - The stress and coping of Israeli emergency room social workers following terrorist attacks. AB - The authors of this qualitative study analyze Israeli hospital social workers' emotional responses to working with civilian casualties in the wake of an unprecedented surge of terrorist violence. Data are based on four focus groups conducted with 38 hospital social workers in relation to their experience with clients in the emergency room. Three themes were identified: (a) Restoring a lost sense of personal security as a necessary stepping-stone toward resuming professional performance, (b) Meeting the families'pain and responding to it, and (c) Disconnecting emotionally in the service of the professional self. The authors discuss the findings in light of the literature on peritraumatic dissociation among helpers. PMID- 15359045 TI - Struggling with recovery: participant perspectives on battling an eating disorder. AB - Recovery from an eating disorder encompasses a complex matrix of mental, physical, and social factors. Although methodological differences affect rates of reported recovery (from 24 to 76%), many women are not experiencing full recoveries from eating disorders. This trend is apparent in persons who discontinue severely destructive behaviors but continue to exhibit psychiatric problems and impairment in social and occupational roles. The purpose of this grounded theory study was to provide an explanatory schema about recovery. Data collection included 17 interviews, a focus group, and participant observation. Findings included repeating wavelike patterns of disease and recovery that seldom returned to a state of normal eating but, instead, remained at a level of atypical eating. PMID- 15359046 TI - Poverty, structural barriers, and health: a Santali narrative of health communication. AB - Recent years have witnessed a surge in scholarship that problematizes the linear, Eurocentric approach to international health communication and suggests the pressing need for a culture-centered approach. This author takes a culture centered approach to exploring the Santali meanings of health in rural Bengal. The open-ended interviews conducted with the Santals bring to surface key issues and meaningful theories of health. Central to the Santali experience of health is food; for the Santal, it is his or her hunger that is the greatest cause of disease and illness. Poverty and the presence of structural barriers that cripple Santali existence emerge as the critical themes of Santali health meanings. The study also illuminates the complex process of meaning making engaged in by participants of marginalized sectors. The author draws policy-based implications from the findings of this research. PMID- 15359047 TI - The role of humor for men with testicular cancer. AB - In this article, the authors examine how 45 men talked about "pure" and "applied" humor in qualitative interviews about their experience of testicular cancer. Most described using applied humor in work and social settings to challenge assumptions about the disease, and in health settings to manage feelings, hide embarrassment, reduce tension, share a sense of solidarity with others, or encourage others to examine themselves. Men also described their usually positive reaction to jokes made by others; jokes helped to dispel tension and reassured men that they were being treated as normal. In a few accounts, men revealed how humor was hurtful. They were sometimes upset about jokes made by others, or by the idea of jokes being made, fearing humiliation and stigma. Humor might ease difficult interactions, but our results suggest that clinicians and others should be careful not to initiate humor without a clear lead from the patient. PMID- 15359048 TI - At the margins? Discourse analysis and qualitative research. AB - Discourse analysis is a qualitative research approach that offers the potential to challenge our thinking about aspects of the reality of health and health care practice. In this article, the author explores one approach to discourse analysis and examines how it offers possibilities for different ways of viewing health and health care practices. She concludes by raising questions as to whether discourse analysis is at the margins of qualitative research, whether that matters, and where discourse analysis might take those margins. PMID- 15359049 TI - Philosophical inquiry in nursing: an argument for radical empiricism as a philosophical framework for the phenomenology of addiction. AB - Philosophical inquiry involves examination of the meaning of a phenomenon within the larger framework of the human condition and a particular view of reality. Argumentation is the method. In this study, the author argues that the metaphysics of radical empiricism enhances the understanding of the immediate subjective experience or phenomenology of addiction. When applied to subjective narratives of the addiction experience from literature, this metaphysics confers meaning to the changes in consciousness that occur in the immediate subjective experience of addiction. Conclusions revealed from the application of this perspective highlight novel feelings, the will and changing sense of self, and the human values of belonging and freedom. This study also revealed the value of the philosophical method of inquiry: to temporize our moral judgments and enrich our perspective of the human values within the addiction phenomenon. These values support and clarify the belief system of nursing. PMID- 15359053 TI - Functional recovery and exercise behavior in men and women 5 to 6 years following coronary artery bypass graft (CABG) surgery. AB - Improvement following coronary artery bypass graft (CABG) surgery has been reported to be less in women than men. Relationships between exercise behavior and functional status of men and women 5 to 6 years after CABG have not been examined in a representative patient sample. This study compared the 5- to 6-year recovery in a cohort of 184 patients at the Minnesota site of the Post CABG Biobehavioral Study. Data were collected by telephone interview and self administered questionnaires. Results showed that women had lower physical ( p 0.05). Ten minutes after CCJ anesthetization, electric waves were recorded from the cecum but not from the CCJ or AC; however, electric activity returned after 90 min. Saline injection did not affect the electric activity of the cecum, CCJ and AC. CONCLUSION: The electric wave parameters of the cecum differed from those of the CCJ and AC, suggesting that the motile activity of the CCJ and AC is not a continuation of the motile activity of the cecum and that it might be evoked by 2 different pacemakers. The similarity in frequency and conduction velocity of electric waves of the CCJ and AC, however, most likely denotes that the AC waves are a continuation of those of the CCJ, and that both are evoked by the same pacemaker probably located in the CCJ. The higher amplitude of cecal waves might be due to the thicker cecal musculature compared to that of the AC. PMID- 15359095 TI - Influence of titanium coating on the biocompatibility of a heavyweight polypropylene mesh. An animal experimental model. AB - INTRODUCTION: In light of the fact that, to date, no information is available about titanium relative to its application in prosthetic material employed for hernial repair, the aim of the present work was to evaluate the fundamental possibilities of titanium-coated polypropylene meshes. MATERIALS AND METHODS: In experiments with animals, two groups, each containing 11 pigs, received either a heavyweight polypropylene mesh (Atrium) or an identical but titanium-coated mesh (titanium-coated Atrium) implanted into the left groin using the totally endoscopic extraperitoneal patchplasty technique. RESULTS: A significant difference in the shrinkage behavior between conventional Atrium and titanium coated Atrium was found (14.9 vs. 8.8%, p < 0.05). Furthermore, the partial volume of the inflammatory infiltrate also proved to be smaller with the titanium coated mesh (14.9 vs. 12.4%). In addition, Ki-67 expression was lower in the group implanted with titanium-coated mesh (21.0 vs. 15.0%). No difference was observed with regard to the apoptosis index (7.6 vs. 6.5). CONCLUSIONS: Heavyweight titanium-coated polypropylene meshes induce a less pronounced foreign body reaction in comparison with identical meshes with no titanium coating, which, since the amount of material implanted is identical, must be attributed solely to the titanium coating. PMID- 15359096 TI - Does mesh implantation affect the spermatic cord structures after inguinal hernia surgery? An experimental study in rats. AB - BACKGROUND: Inguinal hernia repair is the most common operation in general surgery. Prosthetic reinforcement of the inguinal area with polypropylene mesh has increased dramatically in the last decade. The aim of this study was to evaluate how different types of mesh affect the spermatic cord structures. METHODS: Thirty rats were divided into three groups. The spermatic cord was dissected free and a conventional suture repair was performed in group I, an operation mimicking the Lichtenstein operation with a heavyweight polypropylene mesh in group II and the same operation using large pore, lightweighted polypropylene/polyglactin composite mesh in group III. A vasography was performed after 90 days. The cross-sectional area of the vas deferens and s-testosterone from the spermatic vein were measured using the contralateral side as control. Light microscopy of the inguinal canal was performed and inflammation and fibrosis were graded. RESULTS: Vasography revealed patent vas deferens in all animals. In group III, there was a lower s-testosterone in the spermatic vein and a reduced cross-sectional area of the vas deferens on the operated compared to the control side. However, there was no difference in the other groups and there was no significant difference in s-testosterone levels between the groups. There was significantly more inflammation and fibrosis after mesh repair compared to suture repair, but there was no difference between the two mesh groups. Unexpectedly, polyglactin fibres were still seen in specimens in group III after 90 days. CONCLUSION: In conclusion, the only effect on the spermatic cord structures in a rat model is seen as an impaired s-testosterone production and a reduced cross-sectional area of the vas deferens after use of a low-weight composite mesh compared to the control side. No difference in inflammation or fibrosis was found between heavyweight polypropylene mesh and low-weight composite mesh. PMID- 15359097 TI - Construction of a transplantable tissue-engineered artificial peritoneum. AB - BACKGROUND: Peritoneal defects lead to serious postoperative problems. Thus the development of physiological material to cover peritoneal defects is very desirable. AIM: The aim of this study was to develop a transplantable artificial peritoneum. METHOD: The artificial peritoneum consisted of collagen gel, fibroblasts, and mesothelial cells, and histological features were analyzed. The artificial peritoneum at the site of a peritoneal defect in the rat was transplanted to the abdominal wall. RESULTS: Histological examination revealed that the artificial peritoneum consisted of a flat mesothelial monolayer upon a stromal matrix. All transplanted artificial peritoneums adapted well to the host and prevented severe adhesion. CONCLUSION: Our artificial peritoneum may be a useful transplantable bioengineered material for repair of surgical peritoneal defects. PMID- 15359098 TI - Proarrhythmia with non-antiarrhythmics. A review. AB - In 1987, at the American College of Cardiology national meeting, a group of physicians from Europe and the United States agreed to use the term 'arrhythmogenesis' to refer to an aggravation or provocation of arrhythmias resulting from any cause and specifically to use the word 'proarrhythmia' when such arrhythmogenesis is from drug therapy. Proarrhythmia is thus, defined as the potential of cardiac and non-cardiac drugs to induce or exacerbate arrhythmias. It is a relatively common finding in the hospitalized and outpatient settings. It was recognized since the early 1980's, but still was considered an extremely unusual event. In many instances, unfortunately the first manifestation of proarrhythmia is death. We have identified multiple conditions and non-cardiac medications that have been reported in association with this entity. Basic concepts of ion-channels of the heart are provided in this review, to help understanding the rational of the pathophysiology, which remains of paramount importance, as it gives insight to the diagnosis, that is mostly based on electrocardiographic findings. The careful detection of the presence of comordid diseases, makes it possible to prevent, recognize, avoid mistreatment and treat the condition. We present an overview of the cardiac cellular electrophysiology, mechanisms of cardiac arrhythmias and explain the substrates and targets of the pro-arrhythmic actions of non-cardiac drugs. PMID- 15359099 TI - Marital dissolution is followed by an increased incidence of stroke. AB - BACKGROUND: Many studies have reported lower mortality in married people. The relation between marital status and incidence of haemorrhagic and ischaemic stroke is unclear. It is largely unknown whether the risk of stroke is increased the first years after divorce or death of spouse. METHODS: Incidence of first ever stroke (n = 6,184) was followed over 10 years in a cohort consisting of all 40- to 89-year-old inhabitants (n = 118,134) in the city of Malmo, Sweden. Marital dissolution (i.e. divorce or death of spouse) prior to the date of stroke was compared in a nested case-control design (3,134 initially married stroke cases, 9,402 initially married controls). RESULTS: As compared to the married groups, the incidence of stroke was increased in divorced men and women (RR = 1.23, CI: 1.10-1.39 and RR = 1.26, CI: 1.12-1.41, respectively) and widowed men and women (RR = 1.13, CI: 0.99-1.28 and RR = 1.13, CI: 1.02-1.24, respectively) after adjustments for age, country of birth and socioeconomic indicators. The risk of stroke was not increased in never married men. Marital dissolution was followed by increased risk of stroke, which was significant for men (adjusted odds ratio: 1.23, CI: 1.03-1.5) and borderline significant for women below 65 years of age (odds ratio: 1.45, CI: 0.99-2.14). CONCLUSION: The incidence of stroke is increased in divorced and widowed individuals. Never married men do not have an increased incidence. The risk of stroke is elevated during the first years after divorce or death of spouse. PMID- 15359100 TI - Outcome of survivors of acute stroke who require prolonged ventilatory assistance and tracheostomy. AB - BACKGROUND AND PURPOSE: Mechanical ventilation after stroke is associated with high mortality. However, little is known about survivors who require prolonged ventilatory assistance and tracheostomy. Our goal was to assess the rate of pulmonary complication, effect of early tracheostomy and prognosis of patients with stroke requiring prolonged ventilatory support. METHODS: Retrospective review of 97 patients with stroke who required ventilatory assistance and tracheostomy admitted to a single teaching hospital between 1976 and 2000. Outcome was defined using the Glasgow Outcome Scale (GOS). RESULTS: Poor outcome (GOS 1-3) occurred in 74% of patients at 1 year and it was associated with older age (p = 0.03), prior history of brain damage (p = 0.02), and neurological worsening after intubation (p < 0.01). However, long-term functional recovery (GOS 4-5) was possible and more likely after strokes involving the posterior circulation (p = 0.03). Pulmonary complications were prevalent and more frequent before tracheostomy (68 vs. 20% after tracheostomy) but did not determine functional outcome. Mean duration of mechanical ventilation was 11 +/- 19 days and did not significantly differ between outcome groups. Early tracheostomy correlated with shorter ICU and hospital stays (p < 0.01 in both cases). CONCLUSIONS: Surviving patients with stroke who require prolonged ventilatory assistance and tracheostomy can have a better outcome than previously reported. Aggressive care is justified in patients who do not continue to deteriorate neurologically. Pulmonary complications are frequent but treatable. Early tracheostomy can shorten ICU and hospital stays and reduce costs. PMID- 15359101 TI - Clinical implication of cerebral artery calcification on brain CT. AB - BACKGROUND AND PURPOSE: Arterial calcification is a process of atherosclerosis, usually taking place within advanced atheromatous plaque. In the coronary arteries, calcification seen on computed tomography (CT) often indicates severe stenosis, and is noted in almost all patients with coronary symptoms. Calcification of the cerebral arteries is also frequently observed on brain CT, but its clinical significance has yet to be clarified. To evaluate the clinical significance of cerebral arterial calcification on brain CT, we investigated the angiographic and transcranial Doppler ultrasonography (TCD) findings of calcified cerebral arteries. METHODS: The subjects were 57 patients with ischemic stroke, all of whom underwent brain CT, digital subtraction cerebral angiography and TCD. Stroke subtypes were defined according to TOAST classification. Two clinicians who were blinded in respect to the patients' clinical history and to each other's interpretation, determined the presence of cerebral arterial calcification and the degree of angiographic stenosis. RESULTS: Cerebral artery calcification was noted in 23 patients (40%) out of a total of 46 arteries, consisting of 23 internal carotid, 15 vertebral, 5 basilar, 2 middle cerebral and 1 anterior cerebral artery. The patients with calcification were significantly older and more hypertensive than those without calcification. All the patients with calcification showed either large artery atherosclerosis or lacunar stroke. Hemodynamic alterations measured by TCD were rarely observed in calcified arteries. The severity and extent of the atherosclerotic changes were significantly greater in the patients with calcification than those without calcification. CONCLUSION: These results suggest that cerebral arterial calcification does not directly indicate hemodynamically significant atherosclerotic stenosis involving calcified segments, but indicates the underlying etiology of either large artery atherosclerosis or lacunar stroke, and raises the possibility of a diffuse atherosclerotic process being present in the cerebral circulation. PMID- 15359102 TI - Direct dialysis quantification: investigation of the impact of dialysate preservation techniques on solute assays. AB - AIMS: Urease-producing microorganisms may lower urea nitrogen (UN) during dialysate-side dosing. We investigated the impact of 3 proven preservatives (acetic acid, ceftazidime, thimerosal) on UN concentration, and the concentrations of creatinine (CR) and beta2-microglobulin (beta2M). METHODS: The UN, CR and beta2M concentrations were assayed in 3 separate aliquots from 20 spent dialysate samples (ceftazidime, 125 mg/l, or 1% thimerosal, 1 ml/l, vs. control). The beta2M concentration was assayed in 10 further spent dialysate collections (concentrated glacial acetic acid, 5 ml/l, vs. control). Solute concentrations were compared with the concordance correlation coefficient (rc). RESULTS: Ceftazidime and thimerosal had little effect on the concentrations of UN and CR (rc >0.97). For the beta2M concentration, agreement remained good (rc >0.96) for ceftazidime and thimerosal (although the former tended to lower concentrations) but acetic acid was less optimal (rc = 0.893). CONCLUSIONS: Ceftazidime and thimerosal may be used as dialysate preservatives without affecting the UN or CR concentrations. Thimerosal is to be preferred when studying beta2M. Acetic acid produces unacceptable inaccuracy when measuring beta2M. PMID- 15359103 TI - Experimental strategies to reverse chronic renal disease. AB - Progression of chronic nephropathies still represents a major challenge for clinical nephrologists. Specific therapies that prevent patients from requiring dialysis or transplantation are still not available. However, recent experimental studies have demonstrated that regression of advanced lesions in the kidney can be achieved. This review summarizes the recent therapeutic advances using experimental models that might translate into novel human therapies to prevent, or significantly delay, requirement of renal replacement therapy. PMID- 15359104 TI - Efficacy of erythropoietin on dialysis in patients with beta thalassemia minor. AB - BACKGROUND: It is unknown whether chronic erythropoietin (EPO) treatment is able to normalize hemoglobin (Hb) levels and ameliorate cardiac remodeling avoiding blood transfusions in uremic blood transfusion-dependent patients with beta thalassemia minor (beta-thal). METHODS: In 12 hemodialysis (HD) patients with beta-thal, requiring blood transfusions despite EPO therapy, we planned to increase Hb levels up to the target levels (11-12 g/dl) within a one-year period by administering progressively higher doses of EPO (correction phase). We also planned to maintain the Hb target for an additional year (maintenance phase). RESULTS: In the year before the study, patients required 3.3 +/- 0.9 units of packed red blood cells. At baseline, the Hb level obtained with an EPO dose of 212 +/- 73 U/kg/week i.v. was 8.2 +/- 0.8 g/dl. The EPO dose was gradually increased within the first year up to 458 +/- 78 U/kg/week at month 12 (correction phase) and then significantly tapered down during the maintenance phase (390 +/- 54 U/kg/week at month 24). During the correction phase, the Hb levels markedly increased (11.1 +/- 0.3 g/dl at month 12) and did not change in the maintenance phase. No blood transfusion was required throughout the 2 years of follow-up. Left ventricular (LV) mass index progressively decreased from the basal value of 144 +/- 12 to 124 +/- 11 g/m2 in the first year and normalized in all patients at month 24 (109 +/- 12 g/m2, p < 0.001); this occurred in the absence of any change of LV cavity volume index (<90 ml/m2). CONCLUSIONS: In HD transfusion-dependent patients with beta-thal, the administration of high EPO dose for 2 years permits the attainment and the maintenance of Hb targets without blood transfusions. This therapeutic approach permits a complete remission of concentric LV hypertrophy without any adverse effects on the vascular system. PMID- 15359105 TI - Effectiveness of various chemical disinfectants versus cleaning combined with heat disinfection on Pseudomonas biofilm in hemodialysis machines. AB - The development of bacterial biofilms in the hydraulic circuit of hemodialysis machines is routinely prevented by frequent use of a variety of chemical and heat disinfection strategies. This study compared the effectiveness of several chemical disinfectants, commonly used either alone or in combination with a treatment regimen that involved cleaning plus heat disinfection using an in vitro Pseudomonas biofilm model. Effectiveness of these procedures was evaluated using total and viable biomass quantitation and polysaccharide and endotoxin determination. The chemical disinfection procedures were only partially successful in removing all biofilm components. Heat disinfection alone killed viable biofilm bacteria, but did not remove all the biomass components, including endotoxin. The combination of cleaning with citric acid followed by heat disinfection was the most effective in eliminating all biofilm components from the hydraulic circuit of the in vitro model. PMID- 15359106 TI - Age associated alterations in costimulatory and adhesion molecule expression in lupus-prone mice are attenuated by food restriction with n-6 and n-3 fatty acids. AB - Costimulatory and adhesion molecules are known to play a major role in the pathogenesis of systemic lupus erythematosus. Since fish oil and calorie restriction have been reported to attenuate the development of disease in lupus prone (NZBxNZW)F1 mice, the objective of this study was to assess the expression of these key inflammatory molecules in these mice fed diets differing in n-6 and n-3 fatty acid content and fed either food restricted or ad libitum. Age associated increases in the expression of CD28, ICAM-1, and PGP-1 molecules that are involved in the recruitment of inflamed lymphocytes into the kidney were attenuated in mice restricted in food intake. The increase in costimulatory (CD80 and CD86) and adhesion (ICAM-1, PGP-1, LFA-1, and Mac-1) in peripheral blood mononuclear cells was also attenuated by food restriction and to a lesser extent by fish oil alone. Interestingly, amelioration of lupus (laminin expression and proteinuria) correlated with the above beneficial effects and could be seen even in 24-month-old mice. In summary, food restriction and fish oil delay the onset of lupus disease and increase life span in B/W mice by prolonging the maintenance of a youthful immune phenotype. PMID- 15359107 TI - Resistance to development of collagen-induced arthritis in C57BL/6 mice is due to a defect in secondary, but not in primary, immune response. AB - Collagen-induced arthritis (CIA) is a rodent model of human rheumatoid arthritis. Mice of the H-2(q) (DBA/1J) background are highly susceptible to disease whereas mice of the H-2(b) (C57BL/6, B6) background are resistant. To determine why B6 mice are resistant to disease induction, we systematically analyzed T and B cell immune responses in B6 mice, compared to DBA/1J mice, following immunization with bovine type II collagen (CII). We found that both strains showed similar T cell proliferation and cytokine responses and similar levels of anti-CII antibodies (Abs) at day 12 or day 14 of initial immunization (primary immune response), however, those B6 mice that did not develop arthritis showed a significant defect in T cell responses and significantly lower levels of anti-CII Abs following secondary boosting immunization (day 35 of initial immunization, secondary immune response) compared to DBA/1J mice. Our results define for the first time that a defective secondary immune responses in B6 mice leads to the resistance of CIA. PMID- 15359108 TI - Identification of antigen-specific IgG in sera from patients with chronic prostatitis. AB - Antigen-specific vaccines are one of several molecularly targeted approaches under investigation as possible treatments for prostate cancer. Important to the development of vaccines is the identification of appropriate target antigens. We hypothesized that antigens of the prostate might be identified in patients with the chronic prostatitis/pelvic pain syndrome, a syndrome for which an autoimmune pathology has been proposed. Such antigens might represent naturally recognized target antigens of the prostate that could be investigated in the future as prostate tumor antigens. In this report, we used SEREX to identify proteins expressed in a prostate cDNA expression library recognized by IgG from the sera of patients with chronic prostatitis. Candidate proteins were evaluated using a panel of sera from 62 subjects with symptomatic prostatitis and 71 control male blood donors. We identified one protein that was recognized primarily in sera from subjects with prostatitis compared with controls. MAD-PRO-34, a nucleolar autoantigen, was recognized in 6/62 subjects and 0/71 controls (p = 0.00897). This protein had previously been identified as an autoantigen in patients with prostate cancer. In addition, the NY-CO-7 protein was recognized in 9/62 subjects and 3/71 controls (p = 0.0654). Two subjects had IgG specific for both the MAD PRO-34 and NY-CO-7 gene products. Our results demonstrate that some patients with the chronic prostatitis/pelvic pain syndrome have autoantibodies to specific proteins. Proteins identified, and MAD-PRO-34 in particular, could be further investigated as potential prostate tumor antigens. PMID- 15359109 TI - Effects of exogenous IL-2 administration on the homeostasis of CD4+ T lymphocytes. AB - IL-2 is currently used in HIV-infected patients to treat CD4+ T lymphopenia. In order to document a mechanism accounting for its capacity to restore immune function, we studied the effects of IL-2 administration in mice. IL-2 treatment of C57BL/6 mice for 4 days leads to a transient accumulation of CD4+ T lymphocytes. Whereas memory and activated CD4+ T lymphocytes accumulate after IL 2 treatment in both lymphoid and nonlymphoid organs, naive CD4+ T cells only accumulate in the former. IL-2 transiently increases CD4+ T lymphocyte numbers in lymphopenic IL-7(-/-) mice. Studies in T-cell-reconstituted Rag(-/-) gamma c(-/-) mice and in thymectomized mice demonstrated that IL-2 acts directly on peripheral CD4+ T lymphocytes. In vivo labeling of thymocytes showed that IL-2 also stimulates the release of CD4+ thymocytes from the thymus. Therefore, IL-2 treatment acts centrally and peripherally to increase the size of the naive CD4+ T lymphocyte compartment. This dual activity of IL-2 treatment may influence the quality of restoration of this compartment, especially regarding the ability to reconstitute a normal T lymphocyte repertoire. PMID- 15359110 TI - Persistent hypogammaglobulinemia following mononucleosis in boys is highly suggestive of X-linked lymphoproliferative disease--report of three cases. AB - Hypogammaglobulinemia is a common symptom in different immunodeficiencies. It is, however, not usually associated with Epstein-Barr virus (EBV) infections. The X linked lymphoproliferative disease (XLP) on the other hand shows immunological changes in response to the EBV. Here we report three previously healthy boys, all of which developed persistent hypogammaglobulinemia following severe acute infectious mononucleosis. All three patients revealed T-cell abnormalities including inverted CD4/CD8 and increased CD8(+) T-cell numbers. The number of IFN gamma-producing T cells were markedly increased in the two patients studied so far. In addition, patient 2 showed mainly T cells, instead of B cells, to be infected with the EBV. Apart from an uncle of patient 3, who died of malignant lymphoma, family history was unremarkable in all cases. All three patients exhibited mutations in the SH2D1A gene, establishing the diagnosis of XLP. Protein expression was found on immunoblot analysis in one patient with a missense mutation. Development of persistent hypogammaglobulinemia after severe primary EBV infection seems to be a specific diagnostic sign for XLP even in males with unremarkable family history. PMID- 15359111 TI - Regulatory role of vitamin D receptor gene variants of Bsm I, Apa I, Taq I, and Fok I polymorphisms on macrophage phagocytosis and lymphoproliferative response to mycobacterium tuberculosis antigen in pulmonary tuberculosis. AB - The regulatory role of vitamin D receptor (VDR) gene variants of Bsm I, Apa I, Taq I, and Fok I polymorphisms on vitamin D(3)-modulated macrophage phagocytosis with live Mycobacterium tuberculosis and lymphoproliferative response to M. tuberculosis culture filtrate antigen (CFA) was studied in patients with pulmonary tuberculosis (n = 46) and in normal healthy subjects (NHS) (n = 64). Vitamin D(3) at a concentration of 1 x 10(-7) M enhanced the phagocytic potential of normal subjects who had a phagocytic index of less than 20%. This increase was seen in subjects with the genotypes BB (p = 0.017), AA (p = 0.016), tt (p = 0.034), and FF (p = 0.013) and the extended genotype BBAAtt (p = 0.034). Normal subjects with BBAAtt performed better phagocytosis than individuals with bbaaTT genotype (p = 0.034). Vitamin D(3) at 10(-9), 10(-8), and 10(-7) M concentrations suppressed the lymphoproliferative response to CFA antigen in normal subjects. This decreased lymphocyte response was observed in normal individuals with the genotypes BB (p = 0.0009), tt (p = 0.016), and FF (p = 0.008) and the extended genotype BBAAtt (p = 0.02). Addition of vitamin D(3) had no significant effect on macrophage phagocytosis and lymphoproliferative response to CFA in pulmonary TB patients. This may be due to the unresponsive nature of the cells to the action of vitamin D(3) or the downregulated VDR expression by virtue of the disease, which renders them inactive. The genotypes BB, tt, and the extended genotype BBAAtt may be associated with increased expression of VDR which in turn regulate the action of vitamin D(3) and modulate the immune functions to M. tuberculosis in NHS. PMID- 15359112 TI - Blomia tropicalis and Dermatophagoides pteronyssinus mites evoke distinct patterns of airway cellular influx in type I hypersensitivity murine model. AB - Murine models of hypersensitivity to allergens are useful tools for the evaluation of strategies to downmodulate IgE response. We sought to compare allergen inflammatory pulmonary response in previously sensitized mice orally administered with dust mites Blomia tropicalis (Bt) or Dermatophagoides pteronyssinus (Dp). Sensitized A/Sn mice fed with Bt or Dp showed a significant decrease in the IgE response compared to control-immunized mice. Bt-immunized mice demonstrated an accumulation of neutrophils in the bronchoalveolar lavage fluid, while Dp-immunized mice revealed an intense influx of eosinophils in the airway. Bt oral administration did not attenuate cell influx in the airway and Dp fed mice showed a significant decrease of neutrophils and lymphocytes in the bronchoalveolar lavage fluid. These findings demonstrated that oral tolerance induction to Bt and Dp extract in sensitized mice decrease IgE response, but does not interfere in local inflammatory pulmonary response. The distinct profile of airway cellular infiltration between mites immunization suggest an interesting model to study allergic inflammation. PMID- 15359113 TI - Quercetin, a flavonoid phytoestrogen, ameliorates experimental allergic encephalomyelitis by blocking IL-12 signaling through JAK-STAT pathway in T lymphocyte. AB - Experimental allergic encephalomyelitis (EAE) is a Th1 cell-mediated inflammatory demyelinating autoimmune disease model of multiple sclerosis (MS). Quercetin (3,3'4',5,7-pentahydroxy flavone) is a flavonoid phytoestrogen that has profound anticancer and anti-inflammatory activities. In this study, we show that in vivo treatment of SJL/J mice with quercetin (i.p. 50 or 100 microg every other day) ameliorates EAE in association with the inhibition of IL-12 production and neural antigen-specific Th1 differentiation. In vitro treatment of activated T cells with quercetin blocks IL-12-induced tyrosine phosphorylation of JAK2, TYK2, STAT3, and STAT4, resulting in a decrease in IL-12-induced T cell proliferation and Th1 differentiation. These findings highlight the fact that quercetin ameliorates EAE by blocking IL-12 signaling and Th1 differentiation and suggest its use in the treatment of MS and other Th1 cell-mediated autoimmune diseases. PMID- 15359114 TI - Resistance to cytotoxic chemotherapy is induced by NK cells in non-Hodgkin's Lymphoma Cells. AB - It is known that B lymphoma cells are sensitive to cytotoxic chemotherapy, but primary or secondary chemoresistance frequently occurs and is the major cause of death in these patients. However, the mechanisms by which lymphoma cells acquire resistance to cytotoxic drugs are not fully understood. Recently, it was reported that B cells secrete immunoglobulin and produce cytokines after interacting with NK cells, thus indicating the importance of NK/B interactions. In this study, we investigated the mechanism of resistance to cytotoxic chemotherapy induced in cocultures of NK cells and Raji cells. Normally, Raji cells are doxorubicin sensitive, but Raji cells cocultured with NK cells become doxorubicin-resistant. In addition, we detected the upregulation of CD69 and CD70 on Raji cells cocultured with NK cells, suggesting that Raji cells are activated by NK cells. We also found that the resistance of Raji cells to doxorubicin increased when they had been treated with NK cell coculture supernatant. Furthermore, boiled culture supernatant did not inhibit doxorubicin-mediated cell death, indicating that soluble factors are involved. Finally, we confirmed that NK cells produce TNF alpha, and that doxorubicin-sensitive Raji cells become doxorubicin-resistant after TNF alpha treatment. Taken together, these results suggest that B lymphoma cell resistance to doxorubicin-mediated cell death is induced by coculture with NK cells, because of TNF alpha secretion. PMID- 15359115 TI - Antibody response to the Haemophilus influenzae type b-tetanus toxoid conjugate vaccine in healthy and infection-prone individuals with IgG3 subclass deficiency. AB - Searching for a possible explanation for the phenotypic heterogeneity in IgG3 deficiency, we studied the antibody response to a polysaccharide and a protein antigen in IgG3-deficient (IgG3d) adults after vaccination with Haemophilus influenzae type b capsular polysaccharide (Hib CP) conjugated to tetanus toxoid. Distribution of isotypes, idiotypes, clonotypes, and Gm allotypes were compared. All the vaccinated individuals, irrespective of the level of IgG3 and proneness to infections, developed protective levels of anti-Hib CP. Significantly lower prevaccination levels of IgG2 (p < 0.05) and IgG4 anti-Hib CP (p < 0.04 and p < 0.03) were noted among the infection-prone compared to the healthy IgG3d individuals and/or controls. Seventy percent of the IgG3d patients and none of the controls had the low responding Gm(ga-n/ga-n) genotype, while the majority of the controls had the alternative Gm(bfn/bfn) genotype. The conjugate ACT-HIB vaccine efficiently overcomes the IgG3 subclass deficiency state and the genetic predisposition for lower responsiveness, providing protection against Hib and tetanus infections. The proneness to infection in some IgG3d individuals may relate to their low prevaccination antibody levels. PMID- 15359117 TI - Activation and deactivation of gene expression by Ca2+/calcineurin-NFAT-mediated signaling. AB - Ca2+/calcineurin-NFAT-mediated signaling pathways are involved in diverse cellular reactions by regulating gene expression either positively or negatively. The transcriptional activity of NFAT proteins can be either activating or deactivating depending on which binding partners are involved. Interaction of NFAT with AP-1 turns on the genes involved in active immune responses, while NFAT without cooperative binding of AP-1 turns on a T cell anergy program and blocks T cell activation and proliferation. In addition, interaction of NFAT with histone deacetylase (HDAC) proteins induces gene silencing. In this review we focus on the dual function, activator or deactivator, of NFAT and the binding partners that determine the role of NFAT in gene expression. PMID- 15359116 TI - Humoral epitope-spreading following immunization with a HER-2/neu peptide based vaccine in cancer patients. AB - HER-2/neu is a tumor antigen in patients with breast and ovarian cancer. Multiple varieties of vaccine strategies are being developed to immunize patients against HER-2/neu. Studies in animal models have demonstrated both T cell and antibody immunity are needed to mediate an antitumor response. Thirty-five patients, immunized with HER-2/neu peptide based vaccines, were evaluated for the generation of HER-2/neu-specific antibody immunity. Sixty percent of patients developed HER-2/neu IgG specific antibody responses to at least one peptide included in their vaccine. Twenty-nine percent of patients developed IgG immunity to the native HER-2/neu protein after peptide immunization. Humoral intramolecular epitope-spreading within the HER-2/neu protein occurred in 49% of immunized patients. Intermolecular epitope-spreading to p53 was evident in 20% of vaccinated patients. Of those patients who developed new immunity to p53, 71% had demonstrated antibody epitope-spreading within HER-2/neu. PMID- 15359118 TI - Regulation of calcineurin, a calcium/calmodulin-dependent protein phosphatase, in C. elegans. AB - Calcineurin is a calcium/calmodulin-dependent serine/ threonine protein phosphatase. It is a heterodimeric protein consisting of a catalytic subunit calcineurin A, and a regulatory calcium-binding subunit, calcineurin B. The primary sequence of both subunits and heterodimeric structure is highly conserved from yeast to mammals. Calcineurin has long been implicated in various signaling pathways. Calcineurin genes (cna-1/tax-6 and cnb-1) have been identified in the nematode Caenorhabditis elegans, which share high homology with their Drosophila and mammalian counterparts. C. elegans calcineurin binds calcium and functions as a heterodimeric protein phosphatase establishing its biochemical conservation. Calcineurin expresses in diverse tissues implicating its important role in various physiological processes. This review will focus in brief on the expression pattern and regulation of calcineurin including its effect on growth and development, locomotion, egg-laying, and sensory responses. PMID- 15359119 TI - In vitro selection of specific RNA aptamers for the NFAT DNA binding domain. AB - Nuclear factor of activated T cells (NFAT) plays a central role in the immune response, and the immuno-suppressive drugs, cyclosporin A and FK-506, have been developed to inhibit it. However, due to the toxic effects of these drugs, which derive from their ability to inhibit calcineurin in non-immune tissues, the identification of small compounds that target NFAT directly could be an approach to developing less toxic immunosuppressive therapy. Using an in vitro selection technology termed SELEX on a combinatorial RNA library with 40 nucleotide-long random sequences, we have isolated two RNA aptamers to the NFAT DNA binding domain (DBD). Gel retardation assays and surface plasmon resonance measurements showed that the aptamers have a specific and high affinity (apparent KD~10 to 100 nM) for the NFAT DBD. Enzymatic probing analysis showed that the two RNA aptamers have similar structures and share a sequence that forms an apical loop. Moreover, RNase footprinting analysis showed that the shared sequence (GATATGAAGGA/ TGTG/AGAGAG) is critical for binding to both NFATp DBD and NFATc DBD. These results suggest that short RNAs identified in this study is a specific aptamer to NFAT DBD, and hence could be applied not only for the delineation of NFAT functions but for the development of potent immune modulating lead compounds. PMID- 15359120 TI - Solution structure of the cytoplasmic domain of human CD99 type I. AB - Human CD99, which is encoded by the mic2 gene, is a ubiquitous 32 kDa transmembrane protein. Its major cellular functions are related to homotypic cell adhesion, apoptosis, vesicular protein transport, and differentiation of thymocytes or T cells. Recent reports have suggested that expression of a splice variant of CD99 increases the invasiveness of human breast cancer cells. In order to determine the structural basis of CD99 function, we have initiated structural studies on the human CD99 Type I cytoplasmic domain (hCD99cytoI) using circular dichroism and multi-dimensional NMR spectroscopy. The solution structure of hCD99cytoI shows that it has a hairpin shape anchored by two flexible loops. Consequently, hCD99cytoI does not have any regular secondary structural element; however, the NMR and CD data indicate that it possesses an intrinsic helical nature. PMID- 15359121 TI - Nitric oxide synthase and calcium-binding protein-containing neurons in the hamster visual cortex. AB - The distribution and morphology of neurons containing neuronal nitric oxide synthase (NOS), and calcium-binding proteins calbindin D28K and calretinin in the hamster visual cortex were compared by immunocytochemistry. Staining for NOS, calbindin D28K and calretinin was seen both in the specific layers and in the selective cell types. The densest concentration of anti-NOS-immunoreactive (IR) neurons was found in layer VI. Most of the calbindin D28K-IR neurons were located in layers II/III and V while the calretinin-IR neurons were predominantly located in layers II/III. The labeled neurons varied in morphology. The large majority of NOS-IR neurons were round or oval cells with many dendrites coursing in all directions. The majority of the calbindin D28K-IR neurons were stellate and round or oval cells with multipolar dendrites. The majority of the calretinin-IR neurons were vertical fusiform cells with long processes traveling perpendicular to the pial surface. Our study showed that 14.7% and 27.5% of the NOS-IR cells in the hamster visual cortex contained calbindin D28K or calretinin, respectively. These results indicate that NOS, calbindin and calretinin are located in specific layers and specific cell types and the vast majority of NOS-containing neurons are limited to neurons that do not express calbindin D28K or calretinin. PMID- 15359122 TI - Cloning and expression of a lipase gene from rice (Oryza sativa cv. Dongjin). AB - Lipases are useful enzymes that are primarily responsible for the hydrolysis of acylglycerides during lipid processing. We have cloned a lipase gene from a rice seed coat cDNA library (Oryza sativa cv. Dongjin). The cDNA was 1,445 bp in length and encoded 361 amino acid residues (GenBank accession No. AY580163). The deduced amino acid sequence had 82 and 56% identity to Oryza sativa (cv. Chuchung) and Arabidopsis thaliana lipase genes, respectively, and there was a GxSxG consensus motif near the catalytic triad at the active serine site. The deduced sequence had little homology to mammalian and microbial lipases. When the Oryza sativa lipase gene was expressed in Escherichia coli with the pET expression system, activity was found mainly in the pellet fraction. The purified product had lipolytic activity towards tributyrin and was about 40 kDa in size. PMID- 15359123 TI - Aurintricarboxylic acid translocates across the plasma membrane, inhibits protein tyrosine phosphatase and prevents apoptosis in PC12 cells. AB - Aurintricarboxylic acid (ATA) prevents apoptosis in a diverse range of cell types including PC12 cells. It is known to stimulate tyrosine phosphorylation of signaling proteins including Shc proteins, phosphatidylinositol 3-kinase, phospholipase C-g and mitogen-activated protein kinases (MAPKs). However, it has been unclear how ATA increases the phosphorylation of these proteins as it was believed to be membrane impermeable. We found that ATA translocates across the plasma membrane of PC12 cells and have confirmed that it is a potent inhibitor of protein tyrosine phosphatases (PTP ases). Other PTPase inhibitors also prevented apoptosis independent of ATA. These observations indicate that ATA exerts its anti-apoptotic effect on PC12 cells at least in part by inhibiting certain PTPase(s). PMID- 15359124 TI - Analysis of the root nodule-enhanced transcriptome in soybean. AB - For high throughput screening of root nodule-enhanced genes, cDNA libraries specific for three different developmental stages of soybean root nodules were constructed after inoculation with Bradyrhizobium japonicum USDA110. 5,469 cDNA clones were sequenced and grouped into 2,511 non-redundant (nr) ESTs consisting of 769 contigs and 1,742 singletons. Using similarity searches against several public databases we constructed a functional classification of the ESTs into root nodule-specific nodulin genes, stress-responsive genes and genes related to carbon and nitrogen metabolism. We also constructed a cDNA microarray with 382 selected clones that appeared to be up-regulated in the root nodule. Using the microarray we compared the transcript levels of uninfected roots and root nodules from four developmental stages. We identified 81 genes that were differentially expressed, and grouped them into seven clusters according to the similarity of their expression profiles, using a hierarchical clustering algorithm. Clusters 1, 2, 3, and 6, comprised of 58 genes, showed root nodule-enhanced expression. The information from this study will be used to analyze the roles of root nodule specific genes and signaling pathways during root nodule development. PMID- 15359125 TI - Novel and recurrent mutations of the LDL receptor gene in Korean patients with familial hypercholesterolemia. AB - We have identified 16 different mutations of the low-density lipoprotein receptor (LDLR) gene in 25 unrelated Korean patients with heterozygous familial hypercholesterolemia (FH), including five novel mutations, C83Y, 661del17, 1705insCTAG, C675X, and 941-1G>A. The 1705insCTAG mutation in which the four 3 cent -terminal nucleotides of exon 11 are duplicated was found to prevent splicing of exon 11 and would therefore generate a truncated polypeptide. The in frame 36-bp deletion (1591del36) in exon 11, which had been reported only in one Korean FH patient, was also found. We showed that this change affects transport of the LDL receptor from the endoplasmic reticulum to the cell surface. In addition, we found 8 mutations (-136C>T, E119K, E207K, E207X, F382L, R574Q, 1846 1G>A, and P664L) that had been described in other ethnic groups but not in Koreans, and 2 mutations (R94H and D200N) that had been described in Koreans as well as other ethnic groups. 5 mutations (1591del36, E119K, E207X, E207K, and P664L) were found more than once in the Korean FH samples. Identification of the novel and recurring LDLR mutations in Korean FH patients should facilitate prenatal and early diagnosis in families at high risk of FH. PMID- 15359126 TI - Molecular cloning and characterization of a 2-deoxystreptamine biosynthetic gene cluster in gentamicin-producing Micromonospora echinospora ATCC15835. AB - The organization of the 2-deoxystreptamine (DOS) biosynthetic gene cluster of Micromonospora echinospora has been determined. Sequencing of a 14.04 kb-region revealed twelve open reading frames (ORFs): four putative DOS biosynthetic genes (gtmA, B, C, and D), five amino sugars biosynthetic genes (gtmE, G, H, I, and gacB), two aminoglycoside resistance genes (gtmF and J) as well as a hypothetical ORF (gacA). One of the putative DOS biosynthetic genes, gtmA, was expressed in Escherichia coli, and the purified protein was shown to convert glucose-6 phosphate (G-6-P) to 2-deoxy-scyllo-inosose (DOI), a key step in DOS biosynthesis. In addition gtmJ was expressed in Streptomyces lividans and shown to confer gentamicin resistance. Thus gtmA and gtmJ are implicated in the biosynthesis of gentamicin and in resistance to it, respectively. PMID- 15359127 TI - Changes in the reproductive functions of mice due to injection of a plasmid expressing an inhibin alpha-subunit into muscle: a transient transgenic model. AB - Inhibin is a gonadal hormone composed of an a-subunit and one of two beta subunits (betaA, betaB), and its primary role is to inhibit FSH secretion by the pituitary. To investigate the roles of inhibin alpha in the reproductive system, an expression plasmid, pCMV-rINA, with the rat inhibin alpha cDNA fused to the cytomegalovirus promoter, was introduced into muscle by direct injection. Inhibin alpha mRNA was detected in the muscle by RT-PCR and Southern blot analysis. Inhibin protein was also detected, and Western blot analysis revealed a relatively high level of serum inhibin, but not of activin betaA. The estrous cycle of the pCMV-rINA-injected mice was extended, but there was no change in levels of pituitary FSH mRNA or serum FSH and no ovarian cysts were observed. When injected female mice were mated with males of proven fertility, litter size increased. Surprisingly, the embryos of pregnant females injected with pCMV-rINA, were retarded in growth and had defects in internal organs. When male mice were injected, testicle weight increased slightly without any noticeable change in the histology of the seminiferous tubules. Taken together, our data indicate that the inhibin alpha subunit influences a number of the reproductive functions of female mice. PMID- 15359128 TI - The distribution and expression of HERV families in the human genome. AB - The families of human endogenous retroviruses (HERVs) are widely distributed in the human genome. Here we examined their distribution and expression. Approximately forty thousand HERV elements including truncated and solitary long terminal repeats (LTRs) were identified. These elements were most dense on chromosomes 4, 20, X, and Y. From an analysis of genomic stability during primate evolution, the 5 cent -LTR of the HERV genome (5 cent LTR - internal HERV - 3 cent LTR) appeared to be more often truncated than the 3 cent -LTR. ESTs derived from normal placenta, skeletal muscle, hypothalamus, and testis gave frequent matches to HERV elements. We present a classification of genes associated with HERV elements according to the hierarchical structure of gene ontology. PMID- 15359129 TI - Expression analysis of the pistil genes in controlling self-incompatibility of brassica campestris by CO2 gas using microarray. AB - In Brassica campestris, self-incompatibility (SI) can be overcome by CO2 gas treatment. Previously, we reported inhibition of the SI in the Brassica campestris line, 734, by CO2 gas. In this study, we used a cDNA microarray comprising 1,184 unique cDNA ESTs from a pistil-specific cDNA library of the Brassica line to examine the expression of the pistil genes in response to CO2. Microarray analysis revealed that expression of 2% of the pistil genes was altered by exposure to CO2 gas. Surprisingly, the SLG gene of the susceptible line was one of the down-regulated genes. Transmission electron microscopic analysis showed that the CO2 caused morphological changes in the papillary cell. Our results demonstrate that the overcoming of SI by CO2 gas involves reduced expression of the SLG gene in Brassica. PMID- 15359131 TI - Ectopic expression of a cold-inducible transcription factor, CBF1/DREB1b, in transgenic rice (Oryza sativa L.). AB - The gene encoding C-repeat/dehydration-responsive element binding factor 1 (CBF1/DREB1b) of Arabidopsis was introduced into rice (Oryza sativa L.) under the control of the maize ubiquitin promoter. Its incorporation and expression in transgenic rice plants were confirmed by DNA and RNA gel-blot analyses. Cold tolerance in the transgenics was not significantly different from that of the wild-type plants, as determined by ion leakage, chlorophyll fluorescence, and survival rates. However, the cold-responsive genes lip5, lip9, and OsDhn1 were up regulated in the transgenic plants, suggesting that the cold signal transduction pathway involving CBF1 is partially conserved in this cold-labile plant. PMID- 15359130 TI - Chromatin remodeling facilitates DNA incision in UV-damaged nucleosomes. AB - The DNA repair machinery must locate and repair DNA damage all over the genome. As nucleosomes inhibit DNA repair in vitro, it has been suggested that chromatin remodeling might be required for efficient repair in vivo. To investigate a possible contribution of nucleosome dynamics and chromatin remodeling to the repair of UV-photoproducts in nucleosomes, we examined the effect of a chromatin remodeling complex on the repair of UV-lesions by Micrococcus luteus UV endonuclease (ML-UV endo) and T4-endonuclease V (T4-endoV) in reconstituted mononucleosomes positioned at one end of a 175-bp long DNA fragment. Repair by ML UV endo and T4-endoV was inefficient in mononucleosomes compared with naked DNA. However, the human nucleosome remodeling complex, hSWI/SNF, promoted more homogeneous repair by ML-UV endo and T4-endo V in reconstituted nucleosomes. This result suggests that recognition of DNA damage could be facilitated by a fluid state of the chromatin resulting from chromatin remodeling activities. PMID- 15359132 TI - Ginsenosides regulate ligand-gated ion channels from the outside. AB - Treatment with ginsenosides, the major active ingredients of Panax ginseng, produces a variety of physiological effects on the central and peripheral nervous systems. Ginsenosides inhibit various types of ligand-gated ion channel but it is not clear whether they act from within or outside the cell since they are somewhat membrane-permeable. In the present study, we used the Xenopus oocyte gene expression system to determine from which side of the cell membrane the ginsenoside Rg3 (Rg3), and M4, a ginsenoside metabolite, act to regulate ligand gated ion channel activity. Ligand-gated ion currents were measured using the two electrode voltage clamp technique. Rg3 and M4 inhibited 5-HT3A and a3b4 nACh receptor-mediated ion currents when present outside of the cell but not when injected intracellularly. We also examined the effect of these agents on oocytes expressing the gustatory cGMP-gated ion channel, which is known to have a cGMP binding site on the intracellular side of the plasma membrane and is only activated by cytosolic cGMP. Rg3 inhibited cGMP-gated ion currents when applied extracellularly or to an outside-out patch clamp, but not when injected into the cytosol or when using an excised inside-out patch clamp. These results indicate that Rg3 and M4 regulate ligand-gated ion channel activity from the extracellular side. PMID- 15359133 TI - A protein interaction map of soybean mosaic virus strain G7H based on the yeast two-hybrid system. AB - A protein interaction map of Soybean mosaic virus (SMV) strain G7H was generated by the yeast two-hybrid system. Clones encoding the genes P1, HC-Pro, P3, 6K1, CI, 6K2, VPg, NIa, NIb, and CP were fused downstream of the GAL4 binding domain (GAL4-BD) and of the GAL4 activation domain (GAL4-AD). The GAL4-BD and GAL4-AD fusion derivatives of each gene were co-transformed into yeast and transformants in which interaction took place were identified on selective media. Interacting fusion proteins were extracted from the yeast cells, run on SDS-PAGE gels and finally checked by Western blotting with GAL4 polyclonal antibodies. Strong interactions were detected between the pairs CP/CP, HC-Pro/HC-Pro, NIa/NIa, and CP/HC-Pro. Relatively weak but significant interaction was detected between VPg and NIa. Although not all of the protein-protein interactions previously reported in other potyviruses were detected, the interactions revealed here were, in general, similar to those reported previously. PMID- 15359135 TI - Treating hypertension in diabetes patients. PMID- 15359134 TI - Construction of a vector generating both siRNA and a fluorescent reporter: a siRNA study in cultured neurons. AB - RNA interference is an important tool for gene silencing. However, its application to primary cultured cells has been limited by low transfection efficiencies. In this work we developed a vector which encodes both siRNA and red fluorescent protein. Using this vector we could markedly suppress green fluorescent protein (GFP) and bim an endogenous gene. Primary cultured cortical neurons transfected with siRNA against doublecortin showed that doublecortin expression was significantly inhibited in nearly all the transfected neurons. This vector identifies the transfected cells and should be useful for loss-of gene function studies in neurons. PMID- 15359137 TI - Recognizing depression in children. PMID- 15359139 TI - Are you up to date on immunizations? PMID- 15359141 TI - Arterial insufficiency and ulceration: diagnosis and treatment options. PMID- 15359142 TI - The great malpractice debate. PMID- 15359148 TI - Telithromycin (Ketek) for community-acquired respiratory tract infections. PMID- 15359151 TI - Practice branding and implant dentistry. PMID- 15359152 TI - European markets for dental implants and final abutments 2004: executive summary. PMID- 15359153 TI - Repair of fractured framework: scanning electron microscopy and energy dispersive X-ray spectroscopy. AB - Fractured metal prostheses can be analyzed for possible causes of failure using scanning electron microscopy (SEM). In this study, fractography is used to determine the cause of the failure and whether repair is practical. Energy dispersive x-ray spectroscopy (EDS) is used to determine composition of the fractured prosthesis so that a repair process can be recommended. The technique is presented for the repair of a titanium framework for an implant-supported overdenture based on the analysis data. PMID- 15359154 TI - Fracture and displacement of lingual cortical plate of mandibular symphysis following bone harvesting: case report. AB - This article describes a unique complication after chin bone harvesting. The complication consisted of fracture and posterior displacement of the lingual cortical plate that did not occur at the time of the operation but during the healing phase. The bone was harvested for a bilateral sinus lift procedure. Diagnosis was made by chance with the aid of a postoperative CT scan that was taken to study the sinus area. The mandibular scans revealed a bony fragment 2 to 4 mm in width and 3 cm in length fractured and displaced 1 cm posteriorly. This bony fragment was pedicled to the geniohyoid and genioglossus muscles. The patient was asymptomatic, and no treatment was carried out. The patient is still symptom-free 16 months after the initial diagnosis of the fracture. PMID- 15359155 TI - Rationale for the application of immediate load in implant dentistry: Part I. AB - Immediate loading in implant dentistry is increasing in popularity as a clinical procedure. A scientific rationale of immediate occlusal loading of the implant support system should emphasize methods to decrease surgical trauma during implant placement and to decrease bone loading trauma during the early loading period. The surgical trauma may be reduced by decreasing heat generation and pressure necrosis. The early loading trauma may be decreased by decreasing the bone strain adjacent to the implant interface. Greater microstrain conditions in bone increase the remodeling rate of bone. The higher the remodeling rate, the weaker the bone and the more risk of occlusal overload. Occlusal overload may lead to implant failure. Since strain is directly related to stress, methods to decrease stress are beneficial. In the present report, the stress-reducing influences include increasing the number of implants. PMID- 15359156 TI - Removable partial denture design involving teeth and implants as an alternative to unsuccessful fixed implant therapy: a case report. AB - Various dental implants have been used, and high success rates have been reported. However, as their uses increase, implant failures have been reported. There are partially edentulous patients with an initial treatment plan of an implant-supported fixed partial denture. At times, one or more strategic implants fail to osseointegrate. In situations where financial, systemic, or local conditions preclude the use of a fixed partial denture, a well constructed removable partial denture can be an excellent treatment alternative. It has been reported that prosthesis support can be improved with the use of osseointegrated dental implants, with improved preservation and maintenance of existing hard and soft tissues around dental implants. This article describes the prosthodontic rehabilitation of a partially edentulous patient by the use of a removable partial denture design involving teeth and implants as an alternative to unsuccessful fixed implant therapy. This treatment option solved a difficult clinical problem derived from the failure of two strategic implants and provided the patient with an esthetic and functional prosthesis. PMID- 15359157 TI - Guided bone regeneration at immediate implant insertion and loading: a case report. AB - A tapered-end, SLA- (sandblasted and acid-etched) surfaced ITI implant was placed in a compromised ridge at the time of tooth removal. After placement of nonautogenous regenerative materials, the implant was immediately loaded with a provisional restoration within 2 hours of implant insertion. Six-month reentry demonstrated regeneration of lostalveolar bone surrounding the implant and clinical implant immobility. Clinical ramifications of these findings are discussed. PMID- 15359158 TI - 8-year onlay bone graft and ridge augmentation with PepGen P-15: a clinical and radiographic case study. AB - The long-term efficacy of PepGen P-15, a tissue-engineered particulate bone replacement graft, has been established for the treatment of periodontal osseous defects. In this case study, the long-term benefits of PepGen P-15 as a bone replacement graft for onlay grafting and ridge augmentation is evaluated using clinical and radiographic assessments. The radiographic and clinical evaluations continued for a period of over 8 years to confirm the stability of the onlay/ridge augmentation site grafted with PepGen P-15 particulate. PMID- 15359159 TI - Effects of alcohol consumption on osseointegration of titanium implants in rabbits. AB - Alcohol consumption affects bone metabolism by impairing osteoblast proliferation and by increasing osteoclastic activity. The purpose of this study was to evaluate bone formation in alcohol-fed rabbits following the insertion of dental titanium implants. Animals were fed with 20% ethanol sugarcane brandy pre- and postoperatively (group 1), preoperatively only (group 2), and with water as control (group 3). During the postoperative period, rabbits received doses of polyfluorochrome labels (i.e., alizarin, calcein, and tetracycline). Rabbits were killed 8 weeks after the implant insertion. The polyfluorochrome-labeled bone areas in rabbits with alcohol consumption in pre- and postoperative (group 1) and preoperatively only (group 2) were significantly less (P <.05) than in the control group (group 3). The percentage of direct bone-to-implant contact was significantly less in pre- and postoperative (49.5%) and preoperative-only (49.2%) groups than in the control group (64.7%) (P <.05). Alcoholic rabbits demonstrated significantly less bone density and direct bone-to-implant contact. PMID- 15359160 TI - Alcohol intake and osseointegration around implants: a histometric and scanning electron microscopy study. AB - Alveolar wound healing can be modified by local and systemic factors. The aim of this study was to evaluate the possible effect of alcoholic beverage administration (sugarcane brandy) on reparative bone formation around hydroxyapatite/tricalcium phosphate implants inside the alveolar socket. Male Wistar rats had their upper right incisors extracted and the bioceramic granules implanted in the alveoli. The animals received increasing concentrations of brandy until 30 degrees Gay-Lussac was achieved starting 30 days before dental extraction and maintained for periods varying from 1 hour to 6 weeks, until sacrifice. Blood alcohol concentration analysis was performed as well as histological and histometric analysis through light and scanning electron microscopy to examine the relation between alveolar healing components, including new bone trabeculae, and the implants. Blood alcohol concentration was significantly higher in treated animals compared with controls. A significant delay in reparative bone formation was detected in the alveolus of alcoholic rats by a histometric differential point counting method, whereas the presence of the bioceramic in the alveolar socket improved alveolar wound healing in alcohol treated rats. It is suggested that the osteoconductive properties of this bioceramic accelerated alveolar wound healing in alcoholic rats. PMID- 15359161 TI - Screw loosening for standard and wide diameter implants in partially edentulous cases: 3- to 7-year longitudinal data. AB - Screw loosening is considered to be a common problem with both screw-retained and cemented implant restorations. A wider abutment platform, as well as using a torque driver to tighten specifically designed screws may help prevent this loosening. However, there has been no clinical study evaluating either of these. To longitudinally compare the frequency of screw loosening in standard diameter, (3.75 and 4.0 mm) implant supported prostheses to that of wide diameter, (5.0 and 6.0 mm) implant supported prostheses that were hand tightened, and to evaluate whether using a torque driver would minimize or prevent this problem, if screw loosening occurred. A total of 213 dental implants in 106 patients were included in this prospective longitudinal study. Of the implants 68 were wide diameter and 145 were standard diameter implants. Wide diameter implants showed 5.8% screw loosening, while standard diameter implants showed 14.5% screw loosening after insertion with only hand torquing. When these loose screws were tightened with a torque driver, there was no more loosening of screws. Within the limitations of this study, the wide diameter implants tested showed less screw loosening than the standard diameter implants when hand torqued. Additionally, within the scope of our study, using a torque driver to tighten the screws with the recommended force prevented this loosening from reoccurring in all cases. PMID- 15359162 TI - Comparison of the mechanical properties of 2 prosthetic mini-implants. AB - Two prosthetic mini-implants (MTI and MDI), which have very similar shapes, are widely used. In this investigation, the mechanical and physical properties of 2 prosthetic mini-implants were investigated. The flexural properties were measured with a universal testing machine. The surface image was observed by SEM with EDX. X-ray analyses were performed. The maximum strength and proportion limit for the different implants differed significantly (P <0.01); however, elastic modulus did not differ significantly (P >0.01). Although the surface of the MTI was smooth, the MDI had a rough surface. The elemental analysis detected titanium (Ti) in the MTI, and Ti, aluminum, and vanadium in the MDI. From the x-ray diffraction pattern, the MTI, which is composed of pure titanium, had a narrow, sharp Ti (syn) peak, whereas the corresponding peak for the MDI was small and broad. Although the 2 devices have similar shapes and dimensions, their surfaces and mechanical properties differ greatly. MTI is easy to remove and wrought in clinical use, and MDI is excellent in flexural properties compared with MTI. (Implant Dent 2004;13:251-256) PMID- 15359163 TI - SEM of retrieved etched screw implant 3 weeks after placement: a case report. AB - A case report of a retrieved etched screw implant 3 weeks after implant placement is presented. The implant was rinsed and placed in formalin solution and sent to the lab for scanning electron microscope qualitative analysis of the surface coating. Bone fragments and bone matrix could be observed on the etched surface. However, they were not present on the machined portion of the implant. PMID- 15359164 TI - Analysis of the transforming growth factor-beta 1 gene promoter polymorphisms in early osseointegrated implant failure. AB - Transforming growth factor-beta 1 is a multifunctional cytokine involved in extracellular matrix deposition, reduction of inflammation, and promotion of wound healing. Single nucleotide polymorphisms in the promoter region of human transforming growth factor-beta 1 gene, C-509T and G-800A, have been shown to increase the transcriptional activity of this cytokine and have been associated with a variety of diseases. The objective of this study was to investigate the possible association between these single nucleotide polymorphisms and the early implant failure. A sample of 68 nonsmoking patients was divided into two groups: a test group comprising 28 patients with one or more early failed implants and a control group consisting of 40 individuals with one or more healthy implants. Genomic DNA from oral mucosa was amplified by polymerase chain reaction and analyzed by restriction fragment length polymorphism. The significance of the differences in observed frequencies of single nucleotide polymorphisms was assessed using the chi square test and Fisher's exact test. The cited single nucleotide polymorphisms in transforming growth factor-beta 1 were analyzed in combination as haplotype using the computer program ARLEQUIN. The authors did not observe significant differences in the allele and genotypes to both single nucleotide polymorphisms of transforming growth factor-beta 1 gene (C-509T and G 800A) between control and early implant failure groups. The distribution of the haplotypes arranged as allele and genotypes were similar between control and test groups. These results indicate that C-509T and G-800A polymorphisms in the transforming growth factor-beta 1 gene are not associated separately or in haplotype combinations with early implant failure, suggesting that the presence of those single nucleotide polymorphisms alone do not constitute a genetic risk factor for early implant failure in the Brazilian population. PMID- 15359165 TI - Maxillary sinus elevation for implant placement using calcium sulfate with and without DFDBA: six cases. AB - Maxillary sinus lift is a surgical procedure performed to increase the volume of bone mass so that dental implants can be placed in the maxillary arch. Several materials have been suggested to be used for this procedure. The purpose of this study was to present the clinical and histologic results of using calcium sulfate with and without demineralized freeze-dried bone allograft (DFDBA) in sinus lift. Medical-grade sterile calcium sulfate was used alone or in combination with DFDBA in 6 patients undergoing sinus lift surgery for implant placement. Bone biopsies were taken at different times ranging from 6 to 24 months. All samples examined showed bone growth with some possible remnants of the grafted DFDBA. Implants were inserted either at the time of the lift or 6 months later. All of them were secure with primary stability. The cases reported indicate that calcium sulfate can be successfully used alone or in combination with DFDBA for sinus lift procedures and that possible residues of DFDBA can be found within newly generated bone. (Implant Dent 2004;13:270-277) PMID- 15359168 TI - Clinical and financial advantages of moist wound management. AB - Best practice wound management includes moist wound healing and using advanced wound care products that promote efficient use of nursing time. Moist wound healing is the key that enables wounds to heal more rapidly with less pain and less scarring, which increases patient satisfaction while decreasing costs. PMID- 15359169 TI - Ten steps to developing university and agency partnerships. PMID- 15359170 TI - An agency-university research partnership: focus on late-life depression. AB - Does anyone working in home care have the time to do one more thing? We would think most clinicians, administrators, educators, and quality improvement coordinators would answer "NO." This article provides a current example of what happened when three Medicare-certified home health agencies answered "YES" to a research partnership dedicated to addressing the unique challenges of enhancing late-life depression treatment and outcomes in the home care setting. The development of an ongoing home care agency-university partnership is described to stimulate other agencies to consider this type of research collaboration. PMID- 15359171 TI - Coping with stress. AB - Erin felt relieved her job had been preserved during recent downsizing. Unfortunately, the nursing staff was drastically reduced, leaving Erin as the sole wound ostomy continence nurse for the entire agency. Relief at not losing her job eventually evolved into stress due to the increased workload. Erin delegated where feasible in an effort to create manageable control over her responsibilities, yet time stress occasionally remained an issue. A creative, individually centered approach to coping with stress could help Erin to meet this challenge with personal clarity and equanimity. PMID- 15359172 TI - Using the linkage model for integrating evidence into home care nursing practice. AB - A home care agency's patient population with heart failure demonstrated improved outcomes after the agency implemented an evidence-based protocol. The model holds promise for use in other agencies working to improve patient outcomes as well as have an impact on decreasing costs. PMID- 15359173 TI - Oasis M0520 incontinence?: is "leaking" really. AB - OASIS M0520 instructions indicate: If the patient is incontinent AT ALL-even occasional leaking-mark "Patient is incontinent." This article describes why "leaking" is incontinence, various causes of urinary incontinence, and what nurses should do when they discover their patients have this problem. By addressing incontinence in its early stages, the nurse may prevent the patient from developing debilitating incontinence. PMID- 15359175 TI - ICD-9-CM coding changes for home care: effective October 1, 2004. AB - Changes in ICD-9-CM codes for all health providers for 2005 go into effect and are to be implemented on October 1, 2004, with no transition period. This article outlines specific coding changes that may be applicable to home care patients. You'll find the new codes, backgrounds of the disease/condition, and a list of invalid diagnosis codes that can be used as a handy reference for clinicians, managers, and office staff. PMID- 15359176 TI - Implementing a pediatric asthma program: enabling self-care management through education. AB - This article provides an overview of asthma prevalence among children, presents a summary of asthma triggers based on past research, outlines the essential components of a pediatric asthma home care model implemented by a large urban home health agency, and details the types of clinical documentation needed for care plan development and monitoring of asthma in the home. PMID- 15359177 TI - Double trouble: acetaminophen increases the risk of upper GI complications for people taking NSAIDs. PMID- 15359178 TI - Healthcare worker influenza vaccination: personal choice or professional responsibility? PMID- 15359179 TI - A model for home care clinician and home health aide collaboration: diabetes care by nurse case managers and community health workers. AB - Researchers at Johns Hopkins University conducted a randomized clinical trial to test the effect of nurse case management and community health worker interventions on diabetes control among inner city African Americans. The results demonstrated that the greatest improvement occurred when nurse case managers and community health workers worked together. This study has implications for how nurse/home health aide collaboration can enhance diabetes management in home care. PMID- 15359180 TI - Nurses speak out against the marketing of tobacco. PMID- 15359181 TI - Environmental health physics-50 years of progress. AB - Environmental health physics is an interdisciplinary field, involving study of the release, transport, and fate of radioactive material in the environment. Further, it addresses the interaction of humans with radioactive materials within the ambient (outdoor) environment and with the environments associated with modern technology and lifestyles. It also involves both naturally occurring and artificially produced radionuclides with the former generally being by far the highest source of exposure. In fact, doses from naturally occurring radionuclides are increasingly being used as a benchmark for the establishment of dose rate limits for people. Because of the pioneering work of early environmental health physicists, models exist today that can be used to assess the potential impacts of new nuclear facilities prior to their operation. In fact, these people represent the branch of the health physics profession who conducted environmental monitoring programs and performed the associated research studies that led to the identification of the principal radionuclides of interest, the major pathways and mechanisms through which they expose people, and the doses that may result from radioactive materials in the natural and technologically enhanced environments. One of their most important contributions was the identification and quantification of many of the key parameters that serve as input to such models. Monitoring of nuclear weapons development facilities used during and after World War II was the initial stimulus for the establishment of environmental health physics programs. Thereafter, these programs were expanded both nationally and globally, as a result of the atmospheric weapons testing programs of nations such as France, the People's Republic of China, the former Soviet Union, the United Kingdom, and the United States. Additional stimuli were provided by the development of the commercial nuclear power industry. Current environmental programs, particularly within the U.S., focus on decontamination and decommissioning of dormant facilities from these earlier defense and commercial programs. The range of the environmental health physics aspects of these activities is the subject of this paper. Presented at the end of the paper is a summary of some of the more important lessons that have been learned. As will be noted, this is an exciting field that will present challenges to health physicists for years to come. PMID- 15359182 TI - Dose audit and evaluation of work practices during barium procedures using digital radiography techniques. AB - Effective dose and organ doses during barium procedures performed using digital radiography machines were estimated and the related work practices were evaluated. Measured values of dose area product (DAP) were used for the calculation of effective doses. One hundred and thirty eight patients undergoing barium procedures were included in the study. The use of additional 0.2 mm copper filter during barium procedures effectively reduced patient doses. The effective dose during barium swallow procedure varied from 0.03 mSv to 3.5 mSv; during barium meal it varied from 0.18 mSv to 2.62 mSv; and during barium enema it varied from 0.56 mSv to 4.24 mSv. Dose auditing was done on the basis of patient doses, imaging techniques and image quality. Selection of optimized exposure factors imparted lower dose to patients during barium procedures. PMID- 15359183 TI - Radioresistance in mammals induced by low-level chronic irradiation: modeling and experimental investigations. AB - Effects of chronic low-level preirradiation on radiosensitivity of mice is studied experimentally and by making use of the methods of mathematical modeling. The experiments show that a priming exposure to chronic internal irradiation can induce radioresistance in mice. The manifestation of this radioprotection effect is reduced mortality of preexposed specimens after challenge acute irradiation, the reason of the animal death being the hematopoietic syndrome. Therefore, the theoretical investigation of the influence of preirradiation on radiosensitivity of mice is conducted by making use of the mathematical models that describe the dynamics of hematopoietic system in mice exposed to challenge acute irradiation following the chronic one. Modeling results show that the radioprotection effect of chronic low-level long-term (more than 1 mo) preirradiation on mice is caused by decreased radiosensitivity (radioresistance) of the granulocytopoietic system, which appeared at the level of one functional cell pool, blood granulocytes. In turn, the above-indicated state of radioresistance of the granulocytopoietic system is the consequence of hypercompensation of the radiation damage of this system resulting in relaxation to elevated concentrations of blood granulocytes and their nondividing and dividing bone-marrow precursors and in enhanced mitotic activity of the latter. Modeling results also indicate that the radiosensitization effect of chronic low-level short-term (less than 1 mo) preirradiation on mice is due to increased radiosensitivity of lymphopoietic, granulocytopoietic, and erythropoietic systems accompanied by increased or close to the normal level radiosensitivity of thrombocytopoietic system. In turn, increased radiosensitivity of above-noted hematopoietic lines is the consequence of an exhaustion of these cell systems. Modeling estimations of the duration of low-level chronic preexposures, which result in radioprotection and radiosensitization effects on mice, are in agreement with the relevant experimental data. PMID- 15359184 TI - Application of the ICRP clarification of the tritium metabolic model. AB - In 2001, the International Commission on Radiological Protection published a clarification to the model for tritium metabolism. This clarification described the use of the gastrointestinal tract model, respiratory tract model, and transfer compartment in calculations of tritium metabolism. This information was used to derive intake retention fractions for tritium and tritium compounds including tritiated water, organically bound tritium, and tritides. In addition, dose coefficients were derived for tritide compounds including general categories described by the ICRP and some specific compounds. PMID- 15359185 TI - Radiological assessment of petroleum pipe scale from pipe-rattling operations. AB - Petroleum pipe scale, consisting of concentrated inorganic solids such as barium sulfate, can deposit on the inside of down-hole pipes during the normal course of oil field pumping operations. A portion of this scale has been shown to contain naturally occurring radioactive materials (NORM), predominantly compounds of radium. When these pipes are removed from the well, there is a potential for radiation doses to the oil field workers handling the pipes, especially as the pipes are cleaned for reuse. A thorough sampling and measurement protocol was applied under a variety of weather conditions in an outdoor laboratory to obtain an accurate indication of the radiological and aerodynamic characteristics of scale release and dust dispersion during petroleum pipe scale removal from out-of service pipes with a restored, historically relevant outdoor pipe-cleaning machine. Exposure rate data were also obtained for both the pre-cleaned pipes, and the general area inhabited by workers during the descaling operation. Four radiation exposure pathways were investigated: inhalation of pipe scale dust generated during pipe rattling, incidental ingestion of the pipe scale dust, external exposure from uncleaned pipes, and external exposure from pipe scale dispersed on the ground. Pipes from three oil fields were rattled to collect as much industry-representative data as possible. The Ra specific activity of the pipe scale ranged from 33.6 +/- 0.4 to 65.5 +/- 0.7 Bq g, depending on the formation. A median atmospheric dust loading of 0.13 mg m was measured in the operator breathing zone. The respirable fraction was observed to be about 42% to 46%. Based on cleaning 20 pipes per day,250 d per year on average, annual committed effective doses for the operator and helper ranged from 0.11 mSv (11 mrem) to 0.45 mSv(45 mrem) for inhalation and from 19 microSv (1.9 mrem) to 97 microSv (9.7 mrem) for incidental ingestion. Worker annual external dose from the pipe racks ranged from 0 to 0.28 mSv (28 mrem). In the deposition experiment, more than 99% by weight of the deposited scale fell within 2 m of the machine centerline, the vast majority of which was in the downwind direction. The dose from this deposited material dominated the worker dose estimates. The annual external dose from dispersed material was estimated to be 2.8 mSv (280 mrem) for the operator and 4.1 mSv (410 mrem) for the helper. PMID- 15359186 TI - Estimating the dose response relationship for occupational radiation exposure measured with minimum detection level. AB - Occupational exposures are often recorded as zero when the exposure is below the minimum detection level (BMDL). This can lead to an underestimation of the doses received by individuals and can lead to biased estimates of risk in occupational epidemiologic studies. The extent of the exposure underestimation is increased with the magnitude of the minimum detection level (MDL) and the frequency of monitoring. This paper uses multiple imputation methods to impute values for the missing doses due to BMDL. A Gibbs sampling algorithm is developed to implement the method, which is applied to two distinct scenarios: when dose information is available for each measurement (but BMDL is recorded as zero or some other arbitrary value), or when the dose information available represents the summation of a series of measurements (e.g., only yearly cumulative exposure is available but based on, say, weekly measurements). Then the average of the multiple imputed exposure realizations for each individual is used to obtain an unbiased estimate of the relative risk associated with exposure. Simulation studies are used to evaluate the performance of the estimators. As an illustration, the method is applied to a sample of historical occupational radiation exposure data from the Oak Ridge National Laboratory. PMID- 15359187 TI - Room model with three modal distributions of attached radon progeny. AB - In this paper a room model with three modal distributions of attached radon progeny is developed. Recoil factors are recalculated for each of the modes, and different recoil factors than usually used are obtained. Dependence of progeny concentration in various modes on ventilation and attachment rate is presented. Unattached Pb is overestimated up to 15% if one modal distribution is used, which can lead to the overestimation of lung dose. PMID- 15359188 TI - 2003 review of neutron and non-neutron nuclear data. AB - This paper reviews the scientific literature for new measurements of both non neutron and neutron nuclear data. Some of the highlights of this review are the following items. There was a withdrawal of a previous claim for discovery of element 118. There have been new measurements of some isotopic abundance values that have led to changes for many elements. There was a new set of recommended standards for calibration of gamma-ray energies published for many nuclides. There have been new half-life measurements reported for very short lived isotopes, for many long-lived nuclides, and for half-lives of double beta (betabeta) decay measurements for quasi-stable nuclides. There was also a new reassessment reported of spontaneous fission (sf) half-lives for ground state nuclides, which distinguished between the half-lives from sf decay and from cluster decay and from the new cluster-fission decay process. This review reports on various nuclear interactions such as charged particle cross sections (n,p) and (n,alpha) measurements for thermal neutrons incident on light nuclides. New thermal (n,gamma) cross sections and neutron resonance integrals that have been measured or re-evaluated are also presented. PMID- 15359189 TI - Uncertainties in doses due to the number of aerosol particles: study on 239PuO2 using default parameters for workers. AB - The behavior of individual particles has been taken into account to estimate, after inhalation, uncertainties in deposition fractions and doses due to the number of 239PuO2 aerosol particles. Using ICRP models and default parameters recommended for workers, similar results were obtained after calculation and after simulations of exposures to 500 to 3 x 10(6) particles (from about 1 Bq to 6 kBq). Uncertainties obtained from simulations increase as the number of particles decreases. The maximal uncertainties in the deposition fraction were observed for the sequestered regions in which no deposit often occurs for exposures of less than 1 x 10(5) particles. Similar large uncertainties in the number of nuclear disintegrations over 50 years were observed in the sequestered regions as well as in lymph nodes. For a given number of particles, uncertainties in committed equivalent doses gradually decrease from the extra-thoracic region to lymph nodes, bronchi, bronchioles, and alveolar interstitium. For 1 x 10(4) particles, similar uncertainties were observed for the committed equivalent doses delivered to lung, liver, bone surfaces, red marrow and gonads, and for dose per unit intake (DPUI), whereas for 1 x 10(5) particles and more, uncertainties in DPUI were higher than those in equivalent doses. The explanation is that in some simulations the maximal committed equivalent dose was delivered to the extra thoracic respiratory tract region (in which case the DPUI calculation mode must be changed). PMID- 15359190 TI - Sorption of 99mTc radiopharmaceutical compounds by soils. AB - Study of the sorption of 99mTc radiopharmaceutical compounds by soils has assessed the fate of these compounds in the event of a surface spill and examined the potential of these compounds as hydrologic tracers. Sorption from deionized water, filtered Missouri River water, and artificial seawater by five surface soils was investigated. For all water types, the Tc radiopharmaceutical compounds showed greater sorption than the uncomplexed pertechnetate. The most lipophilic complexes showed the highest sorption on soils. PMID- 15359191 TI - A new technique for preliminary estimates of TRU activity on air sample filters and radiological smears. AB - In most nuclear facilities, fixed air samplers and sometimes portable air samplers are used where some probability of a release exists but is not expected, and so the added expense and effort of using a continuous air monitor is not deemed justified. When a release is suspected, naturally occurring radioactive material buildup on the filter typically prevents any quantitative measurements within the first day or so. Likewise, outdoor air measurements suffer from the same limitations (such as those taken during the Los Alamos fires) and so any rapid quantifiable measurements of fixed air sampler/portable air sampler filters which are technically defendable (even though conservative) are of use. The technique presented here is only intended for use in routine health physics survey applications and does not presently appear to be appropriate for sub pico Curie activity determinations. This study evaluates the utility of using a portable continuous air monitor as an alpha spectrometer to make transuranic activity determinations of samples using both the built in algorithm for air monitoring and a simple region of interest analysis. All samples evaluated were from air sample filters taken using a portable air sampler. Samples were taken over many months to quantify effects from natural variation in radon progeny activity distributions. PMID- 15359196 TI - A description of emergency department-related malpractice claims in The Netherlands: closed claims study 1993-2001. AB - BACKGROUND: The aim of this study was to assess the quality of care provided at emergency departments (ED) in the Netherlands by analysing medical liability insurance claims. METHODS: A retrospective study performed by reviewing records at MediRisk, presently the largest insurer for medical liability in the Netherlands. The following data were abstracted from the files available for analysis: medical discipline involved, physician involved (resident or consultant), nature and gravity of the complaint, and final claim disposition. RESULTS: Between 1993 and 2001 a total of 326 claims involving the ED were filed at MediRisk. Of these, 256 claims (79%) were closed and were available for analysis. Medical liability claims were filed primarily for alleged errors in diagnosis and treatment. The majority of claims involved minor surgical conditions: fractures, luxations (joint dislocations), wounds and tendon injuries (210/256, 82%). Residents were involved in 76% of the claims; resident supervision by a consultant was documented in only 15% of the medical records. Permanent patient disability resulting from improper ED treatment was alleged in 22% of the claims. Four per cent of the claims involved the death of a patient. Physicians accepted liability in 16% of the claims filed. Indemnity payments during the 8-year study period totalled Euros 504,000. CONCLUSION: The number of medical liability claims is low compared with the number of patients treated in ED in the Netherlands. Claims primarily concerned alleged mistakes in diagnosis and the treatment of minor trauma. Residents were involved in the majority of the claims. More resident supervision is needed, as are specific training programmes for emergency physicians. PMID- 15359197 TI - Urinary acetoacetate or capillary beta-hydroxybutyrate for the diagnosis of ketoacidosis in the Emergency Department setting. AB - OBJECTIVES: We compared the semiquantitative measurement of acetoacetate using urinary dipsticks with the bedside quantitative fingerprick measurement of the principal ketone bodies 3-beta-hydroxybutyrate, for the diagnosis of ketoacidosis. METHODS: This is a one year retrospective study of patients who presented with hyperglycemia levels of 250 mg/l or greater in the Emergency Department setting. We compared the sensitivity, specificity, and predictive value of ketonuria and ketonemia for the diagnosis of ketoacidosis (urine or blood ketone bodies, blood bicarbonates <20 mmol/l, anion gap >16 meq/l) in a sample of patients for whom the levels of ketone bodies in the blood and urine as well as serum electrolytes were available. RESULTS: We studied 355 hyperglycemic patients. The median time between arrival and dipstick testing was 21 min, and was greater than 2 h in more than 10% of cases. Comparison between ketonuria and ketonemia was performed in 173 patients (6% with diabetic ketoacidosis). Ketonuria equal to or less than one cross or a 3-beta-hydroxybutyrate value lower than 3 mmol/l enabled ketoacidosis to be excluded (negative predictive value 100%). At two-cross cutoff points for ketonuria and at the 3 mmol/l cutoff point for ketonemia, the two tests had the same sensitivity (100%), but the specificity of 3-beta-hydroxybutyrate (94%) was significantly higher (P<0.0001) than that of ketonuria (77%). The best positive predictive value for ketonemia was obtained at the 5 mmol/l cutoff point (100%) and for ketonuria at the three-cross cutoff point (26%). At the three-cross cutoff point for ketonuria and at the 5 mmol/l cutoff point for ketonemia, the two tests had the same negative likelihood ratio (0.1), but the positive likelihood ratio of 3-beta-hydroxybutyrate (infinite) was higher than that of ketonuria. CONCLUSION: The measurement of 3-beta hydroxybutyrate in capillary blood is faster and more effective than the use of dipsticks in the urine to detect ketoacidosis in the Emergency Department setting. PMID- 15359198 TI - An artificial neural network ensemble to predict disposition and length of stay in children presenting with bronchiolitis. AB - BACKGROUND: Artificial neural networks apply complex non-linear functions to pattern recognition problems. An ensemble is a 'committee' of neural networks that usually outperforms single neural networks. Bronchiolitis is a common manifestation of viral lower respiratory tract infection in infants and toddlers. OBJECTIVE: To train artificial neural network ensembles to predict the disposition and length of stay in children presenting to the Emergency Department with bronchiolitis. METHODS: A specifically constructed database of 119 episodes of bronchiolitis was used to train, validate, and test a neural network ensemble. We used EasyNN 7.0 on a 200 Mhz pentium PC with a maths co-processor. The ensemble of neural networks constructed was subjected to fivefold validation. Comparison with actual and predicted dispositions was measured using the kappa statistic for disposition and the Kaplan-Meier estimations and log rank test for predictions of length of stay. RESULTS: The neural network ensembles correctly predicted disposition in 81% (range 75-90%) of test cases. When compared with actual disposition the neural network performed similarly to a logistic regression model and significantly better than various 'dumb machine' strategies with which we compared it. The prediction of length of stay was poorer, 65% (range 60-80%), but the difference between observed and predicted lengths of stay were not significantly different. CONCLUSION: Artificial neural network ensembles can predict disposition for infants and toddlers with bronchiolitis; however, the prediction of length of hospital stay is not as good. PMID- 15359199 TI - A validated clinical model to predict the need for admission and length of stay in children with acute bronchiolitis. AB - OBJECTIVE: To develop and validate a logistic regression model to predict need for admission and length of hospital stay in children presenting to the Emergency Department with bronchiolitis. SETTING: Two children's hospitals in Dublin, Ireland. METHODS: We reviewed 118 episodes of bronchiolitis in 99 children admitted from the Emergency Department. Those discharged within 24 h by a consultant/attending paediatrician were retrospectively categorized as suitable for discharge. We then validated the model using a cohort of 182 affected infants from another paediatric Emergency Department in a bronchiolitis season 2 years later. In the validation phase actual admission, failed discharge, and age less than 2 months defined the need for admission. RESULTS: The model predicted admission with 91% sensitivity and 83% specificity in the validation cohort. Age [odds ratio (OR) 0.86, 95% confidence interval (CI) 0.76-0.97], dehydration (OR 2.54, 95% CI 1.34-4.82), increased work of breathing (OR 3.39, 95% CI 1.29-8.92) and initial heart rate above the 97th centile (OR 3.78, 95% CI 1.05-13.57) predicted the need for admission and a longer hospital stay. CONCLUSION: We derived and validated a severity of illness model for bronchiolitis. This can be used for outcome prediction in decision support tools or severity of illness stratification in research/audit. PMID- 15359200 TI - Predicting the result of our X-rays, a way to identify overuse? the Ulster Hospital Dundonald. AB - OBJECTIVE: The objectives of the study were to identify areas of the unnecessary use of diagnostic plain X-rays in the emergency department. Requests for inappropriate plain X-ray investigations have resource, radiation and overcrowding implications for the emergency department. METHODS: The junior doctor or nurse practitioner treating patients attending the emergency department requiring X-ray completed a proforma prospectively. Questions included the type of X-ray, predicted result of the X-ray and observed result of X-ray. RESULTS: A total of 226 out of 280 X-rays (81%) were accurately predicted, 94 X-rays (34%) were abnormal. Facial bones (100%) and skull (79%) were most commonly X-rayed when the result was predicted accurately to be normal. The threshold for X-raying abdomen, kidney, ureter and bladder and lumbar spine was low with normal accurate predictions of 67, 75 and 75%, respectively. Normal knee and foot X-rays were predicted accurately in 61 and 60% of cases, respectively, with a yield of 28 and 13%, respectively. CONCLUSION: The training of emergency medicine staff should include education about clinical indications from guidelines and recommendations about X-ray use, information on the dose of radiation exposure, implications of resources and overcrowding of departments. PMID- 15359201 TI - A review of tissue glue use in facial lacerations: potential problems with wound selection in accident and emergency. AB - OBJECTIVES: The purpose of this study was to review the types of facial lacerations for which tissue glue was used as a closure method and assess whether current evidence was being followed. METHODS: A retrospective analysis of facial lacerations presenting to an adult Accident and Emergency Department was made over a 6-month period. RESULTS: Out of 200 facial lacerations, 45 were closed using tissue glue. The mean length of the wounds was 2.05 cm, with a range of 0.5 6 cm; 42 were linear and three were non-linear. Senior house officers closed 16 wounds, middle grade doctors closed 19, emergency nurse practitioners closed seven, and consultants closed three. Six cases were closed against current evidence (13%). This included three lacerations that were non-linear and three lacerations greater than 4 cm in length. Four out of six of these cases were closed by senior house officer grades (80%). CONCLUSION: The lack of clarity over the use of tissue glue for facial wounds may be attributable to a lack of awareness and training, and the misinterpretation of randomized trials. Greater awareness is needed of the role of tissue glue, especially among senior house officers. PMID- 15359202 TI - Bam: a university for mankind. PMID- 15359203 TI - A novel case of numb bum. AB - Cocaine is a drug with a myriad of pharmacological properties enabling it to be used as a therapeutic agent in medicine. The adverse effects of the recreational use of this drug are often observed in patients presenting to the Emergency Department. This paper describes a patient who presented with unusual symptoms after the rectal administration of cocaine, and the proposed explanation for these symptoms. PMID- 15359204 TI - Isolated myocardial contusion in blunt chest trauma. AB - The objective of this case report was to document a rare case of isolated myocardial contusion in the setting of blunt thoracic trauma. Although demonstrated by electrocardiogram and myocardium-specific enzymes, the trauma had no clinical relevance and the patient was discharged uneventfully from the intensive care unit. The clinical significance of blunt myocardial contusion is then discussed. PMID- 15359205 TI - A case of unilateral re-expansion pulmonary oedema successfully treated with non invasive continuous positive airway pressure. AB - Unilateral re-expansion pulmonary oedema is a rare threatening complication of the treatment of lung atelectasis, pleural effusion or pneumothorax, the pathogenesis of which is not completely known. The clinical picture varies considerably from asymptomatic radiological findings to dramatic respiratory failure with circulatory shock. There are few literature reports of the treatment of re-expansion pulmonary oedema with non-invasive continuous positive airway pressure. We present the case of a 75-year-old man who presented in our emergency room with a large left-sided spontaneous pneumothorax and developed severe respiratory failure and circulatory collapse after drainage via a chest tube. The diagnosis of unilateral re-expansion pulmonary oedema was made and he was successfully treated with non-invasive continuous positive airway pressure. Literature data about the aetiological and pathogenetic factors of the condition are also considered. PMID- 15359206 TI - Easily reversible hypoxemia and hypotension induced by nimodipine. AB - Calcium antagonists are drugs commonly prescribed for the treatment of hypertension, angina pectoris, cardiac arrhythmias and other disorders because of their efficacy and tolerability. Nevertheless, overdosage and intoxication are well documented. In this paper we report a case of nimodipine overdosage resulting in prolonged hypotension and hypoxemia, which was successfully treated with calcium gluconate. PMID- 15359207 TI - Intentional overdose of Large Animal Immobilon. AB - We describe a case of voluntary self-injection with Large Animal Immobilon, a veterinary anaesthesia product containing etorphine, a very strong opioid, and acepromazine, a phenothiazine. This resulted in cardiorespiratory arrest and the need for sustained haemodynamic support after resuscitation. Large Animal Immobilon is used under specific conditions only, mainly in zoo and wildlife medicine. Primary toxicological analysis, although guided by the presumed toxin, could only detect a metabolite of acepromazine in the urine. Further analysis was able to show some traces of etorphine. A number of topics are treated, including the apparent potency of the etorphine and the selection of a suitable antidote, taking into account the different properties of the respective agents. PMID- 15359208 TI - Polydipsia as another mechanism of hyponatremia after 'ecstasy' (3,4 methyldioxymethamphetamine) ingestion. AB - Acute symptomatic hyponatremia after ecstasy (3,4 methyldioxymethamphetamine; MDMA) ingestion is well documented and has been attributed to the syndrome of inappropriate antidiuretic hormone (SIADH). We report the case of an 18-year-old woman who took five tablets of ecstasy in a suicide attempt and drank 1700 ml water at the Emergency Department (ED). The laboratory findings obtained 5 h after ingestion showed a serum sodium concentration of 130 mmol/l, plasma osmolality of 264 mOsm/kg, urinary osmolality of 335 mOsm/kg and natriuresis of 101 mmol/l. The plasma arginine vasopressin level by radioimmunoassay was 33.7 pmol/l 5 h after ingestion. A gas chromatography-mass spectrometry assay confirmed MDMA in blood samples, with serum concentrations of 0.87 mg/l on arrival. This case report strongly suggests that MDMA reduces serum sodium levels through the dual pathways of SIADH and polydipsia. Accordingly, we believe that hyponatremia may be prevented in ED patients after MDMA ingestion by the early restriction of water intake. PMID- 15359209 TI - Acute pancreatitis caused by amiodarone. PMID- 15359210 TI - Retreatment of chronic hepatitis C virus infection. AB - Despite advances in antiviral therapy for chronic hepatitis C, approximately half of patients undergoing initial treatment fail to achieve a sustained virologic response (SVR), thus prompting consideration of retreatment with alternative regimens. The decision to re-treat should be based on the severity of liver disease, as well as the presence of clinical and virologic predictors of a successful outcome of additional therapy. Retreatment of patients who were prior nonresponders to interferon monotherapy with interferon plus ribavirin results in SVR rates of 13% to 15%, which can be increased to 25% to 40% if peginterferon plus ribavirin is used. Retreatment of patients who were prior nonresponders to interferon plus ribavirin with peginterferon plus ribavirin unfortunately achieves SVR rates of approximately 10%. The growing number of patients who have been treated and have failed initial therapy highlights the need for the development of more efficacious antiviral agents for the treatment of chronic hepatitis C. PMID- 15359211 TI - Controversies with aminosalicylates in inflammatory bowel disease. AB - Aminosalicylates have been shown to exhibit a wide range of anti-inflammatory and immunomodulatory properties. Since the discovery of sulfasalazine's efficacy in ulcerative colitis and the subsequent development of sulfa-free mesalamine delivery systems, aminosalicylates have evolved to become an integral part of our therapeutic armamentarium and are now first-line therapies for the treatment of mildly to moderately active inflammatory bowel disease and for maintenance of remissions after successful induction therapy. Despite the substantial body of evidence supporting the use of aminosalicylates in ulcerative colitis and Crohn's disease, gaps in our evidence base and controversies surrounding aminosalicylates clinical application have emerged. In this review, issues of dose response and optimization of the treatment regimen in ulcerative colitis, the discrimination between oral mesalamine formulations in left-sided colitis, and their efficacy in active and quiescent Crohn's disease are discussed. PMID- 15359212 TI - Endpoints for the assessment of response to gastroesophageal reflux disease therapy--what are the appropriate measures of "success"? AB - Therapeutic efficacy for gastroesophageal reflux disease (GERD) has been justified by a variety of different endpoints. Across the surgical, endoscopic, and pharmacologic treatment interventions, an attempt has been made to justify the success of the effect by objective and subjective means. This has included objective measures of esophageal sphincter pressure, intraesophageal acid exposure, and endoscopic esophagitis. More subjective measures have included symptom response as determined by questionnaires, severity scales, physician assessment, and quality-of-life impact. Despite the innumerable studies reporting various treatment interventions for GERD, overall there is a general lack of a standardized methodology to allow comparison of the relative success achieved among various methodologies. Furthermore, there is a striking lack of use of validated instruments to accurately assess treatment effect in many of these studies. This review focuses on the questions that should be raised by clinicians in their evidence-based evaluation of the outcomes achieved in these GERD intervention trials. PMID- 15359214 TI - Updated guidelines for reprocessing gastrointestinal endoscopies. PMID- 15359215 TI - Indicators for chronic disease surveillance. AB - Chronic diseases account for seven of the 10 leading causes of death in the United States, including the three leading causes of preventable death (tobacco use, improper diet and physical inactivity, and alcohol use). Seventy percent of health-care costs in the United States are for chronic diseases. In 1999, to allow public health officials to uniformly define, collect, and report chronic disease data, the Council of State and Territorial Epidemiologists released Indicators for Chronic Disease Surveillance. The report provided standard definitions for 73 indicators developed by epidemiologists and chronic disease program directors at the state and federal level. The indicators were selected because of their importance to public health and the availability of state-level data. This report describes the latest revisions to the chronic disease indicators. The revised set of 92 indicators includes 63 indicators that were unchanged from the first edition, six that have been revised, and 23 that are new. Four indicators from the first edition were deleted. Of the indicators, 24 are for cancer; 15 for cardiovascular disease; 11 for diabetes; seven for alcohol; five each for nutrition and tobacco; three each for oral health, physical activity, and renal disease; and two each for asthma, osteoporosis, and immunizations. The remaining 10 indicators cover such overarching conditions as poverty, education, life expectancy, and health insurance. Additional information regarding the indicators for chronic disease surveillance is available at http://www.cdc.gov/nccdphp/cdi. PMID- 15359216 TI - Contribution of Kir4.1 to the mouse electroretinogram. AB - PURPOSE: The electroretinogram (ERG) represents the combination of several distinct cellular processes and conductances. Here, we define the contribution that K+ conductance through Kir4.1 channels makes to the mouse ERG. METHODS: To obtain mice expressing different levels of Kir4.1, we mated Kir4.1+/- mice and used PCR to identify Kir4.1+/- and Kir4.1+/+ littermates. In addition, we mated Kir4.1+/- males with females homozygous for the nob (no b-wave) defect, which eliminates post-receptoral contributions to the ERG. After overnight dark adaptation, mice were anesthetized and ERGs were recorded to 7 min stimuli, to focus on slow ERG components, or to strobe flash stimuli, to examine earlier ERG components. RESULTS: The amplitudes of the ERG c-wave and the fast oscillation, measured from the c-wave peak, were significantly larger in Kir4.1+/- mice than in Kir4.1+/+ littermates. In comparison, the amplitude of the light peak, the other main component generated by the retinal pigment epithelium in response to light, did not differ between Kir4.1+/- and Kir4.1+/+ mice. The amplitude of slow PIII, revealed by the nob genetic background, was reduced in Kir4.1+/- mice. CONCLUSIONS: These results indicate that a cornea-negative potential, generated by Kir4.1, normally opposes a positive polarity conductance that is generated by the apical membrane of the retinal pigment epithelium to form the c-wave measured at the corneal surface. PMID- 15359217 TI - Comparative gene expression analysis of murine retina and brain. AB - PURPOSE: Several high-throughput studies have described gene expression in the central nervous system (CNS), and recently there has been increasing interest in analyzing how gene expression compares in different regions of the CNS. As the retina is often used as a model system to study CNS development and function, we compared retina and brain gene expression using microarray analyses. METHODS: Mouse retina, brain and liver RNA was hybridized to a custom cDNA microarray containing 5,376 genes and ESTs, and the data from the quantified scanned images were analyzed using Bioconductor and SAM. Preferential retina expression was confirmed by real-time PCR. The cellular distribution of genes newly identified as retina enriched genes was determined by immunohistochemistry. RESULTS: Using stringent statistical analyses we identified 733 genes that were preferentially expressed in retina and 389 in brain. The retina-liver hybridizations identified an additional 837 retina enriched genes. The cellular distribution in the retina was determined for two genes that had not previously been reported to be expressed in the retina, the transcription regulatory proteins EWS and PCPB1. Both proteins were found primarily in the inner nuclear layer. Finally, a comparison of the microarray data to publicly available SAGE and EST library databases demonstrated only limited overlap of the sets of retina enriched genes identified by the different methodologies. The preferential retinal expression of a subset of genes from the microarray, which were not identified as differentially expressed by other methods, was confirmed by quantitative PCR. CONCLUSIONS: The finding of differences in the groups of identified retina enriched genes from the various profiling techniques supports the use of multiple approaches to obtain a more complete description of retinal gene expression. Characterization of gene expression profiles of retina and brain may facilitate the understanding of the processes that underlie differences between the retina and other parts of the central nervous system. PMID- 15359218 TI - Expression of cytochrome P4501b1 (Cyp1b1) during early murine development. AB - PURPOSE: To examine the embryonic expression of cytochrome P4501b1 (Cyp1b1) gene by whole mount in situ hybridization. METHODS: FVB/NcrlBR mouse embryos staged at 9.5, 10.5, and 11.5 dpc were obtained by timed breeding experiments. Antisense and sense RNA probes labeled with digoxigenin UTP were generated by in vitro transcription of an 848 bp Cyp1b1 cDNA fragment that was subcloned into transcription vector pCRII-TOPO. The digoxigenin labeled RNA was localized using an alkaline phosphatase conjugated anti-digoxigenin Fab fragment. Colorimetric detection of the digoxigenin labeled probe was performed with substrate solution containing 4-nitro-blue tetrazolium chloride (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP). RESULTS: During early stages of murine development Cyp1b1 mRNA was detected in the developing eye, hindbrain, branchial arches, forelimb bud, ligaments supporting the liver primordium and developing kidney. In the eye and forelimb bud Cyp1b1 displayed restricted expression along the axes of development. In the developing eye Cyp1b1 exhibited dorsal expression with respect to the dorso-distal/proximo-ventral axis and anterior expression with respect to the anterior-nasal/posterior-temporal axis. In the forelimb bud Cyp1b1 expression was localized posteriorly. The polarity of Cyp1b1 expression was lost at 11.5 dpc, at which time expression was additionally seen in ventral (eye) and anterior (forelimb bud) areas. CONCLUSIONS: The spatio-temporal expression patterns observed in this study suggest that during early stages of murine development, Cyp1b1 participates in establishment and/or maintenance of polarity along the axes of embryonic development. Expression of Cyp1b1 in the dorso-distal end of the optic cup, from which the ciliary body and iris are derived, correlates with the expression patterns seen in adult tissues and the abnormal development of these structures as part of the glaucoma phenotypes resulting from Cyp1b1 mutations. PMID- 15359219 TI - Downregulation of retinal GLUT1 in diabetes by ubiquitinylation. AB - PURPOSE: To investigate the effect of chronic hyperglycemia on the levels of the glucose transporter GLUT1 in retina and its ubiquitinylation. METHODS: Two diabetic animal models (Goto Kakizaki rats and alloxan-induced diabetic rabbits) and retinal endothelial cells in culture were used. GLUT1 content was determined by western blotting. Glut1 mRNA was determined by RT-PCR and northern blotting. Ubiquitin conjugates were evaluated by western blot analysis. In vitro ubiquitin conjugation activity was evaluated in supernatants using radiolabeled ubiquitin. Evidence for GLUT1 ubiquitinylation was further investigated by transfecting HEK293 cells with a hemagglutinin (HA)-tagged ubiquitin cDNA followed by immunoprecipitation of the cell lysates. RESULTS: Chronic hyperglycemia resulted in a significant decrease on the amount of GLUT1 protein without significant changes on the GLUT1 mRNA in the retinas of diabetic GK rats and alloxan treated rabbits, and in high glucose treated retinal endothelial cells, compared to controls. The content of high molecular weight ubiquitin conjugates was higher both in the membrane fractions of diabetic retinas and in endothelial cells incubated with high glucose concentrations. GLUT1 immunoprecipitated from diabetic retinas crossreacted with antibodies directed against ubiquitin suggesting that GLUT1 is posttranslationally modified by monoubiquitinylation. Cells transfected with HA-tagged ubiquitin revealed crossreactivity with anti GLUT1 antibodies on the HA immunoprecipitates. CONCLUSIONS: The data indicate that retinal GLUT1 abundance decreases in experimental diabetes and with exposure of retinal endothelial cells to elevated glucose concentrations. Results further suggest that decreased abundance of GLUT1 may be associated with its increased degradation by a ubiquitin dependent mechanism. Ubiquitinylation of GLUT1 may be the mechanism targeting GLUT1 for degradation in diabetes. PMID- 15359220 TI - Changing our name: for pain management nursing. PMID- 15359221 TI - A pilot study of the effectiveness of guided imagery with progressive muscle relaxation to reduce chronic pain and mobility difficulties of osteoarthritis. AB - Osteoarthritis (OA) is a common, chronic condition that affects most older adults. Adults with OA must deal with pain that leads to limited mobility and may lead to disability and difficulty maintaining independence. A longitudinal, randomized clinical trial pilot study was conducted to determine whether Guided Imagery (GI) with Progressive Muscle Relaxation (PMR) would reduce pain and mobility difficulties of women with OA. Twenty-eight older women with OA were randomly assigned to either the treatment or the control group. The treatment consisted of listening twice a day to a 10-to-15-minute audiotaped script that guided the women in GI with PMR. Repeated-measures ANOVA revealed a significant difference between the two groups in the amount of change in pain and mobility difficulties they experienced over 12 weeks. The treatment group reported a significant reduction in pain and mobility difficulties at week 12 compared to the control group. Members of the control group reported no differences in pain and non-significant increases in mobility difficulties. The results of this pilot study justify further investigation of the effectiveness of GI with PMR as a self management intervention to reduce pain and mobility difficulties associated with OA. PMID- 15359222 TI - Gender differences in chronic pain--findings from a population-based study of Norwegian adults. AB - The purpose of this study was to evaluate for gender differences in the experience of chronic pain and the impact of chronic pain on quality of life (QOL). A total of 1,912 out of 4,000 Norwegian citizens returned a mailed questionnaire, and 24.4% reported chronic pain. More women than men reported chronic pain, and women reported significantly higher pain intensity scores than men did. Although the duration of chronic pain was similar in women and men, women more often received treatment for their chronic pain. Men in chronic pain reported a poorer QOL than women did. Although specific variables that predicted present pain intensity in women and men differed, the variables that explained the largest percentage of the variance in pain for both genders were the disease and the pain location variables. These findings support previously published studies that document gender differences in chronic pain and extend the work to the impact of chronic pain on QOL. PMID- 15359223 TI - The pain experience of cognitively impaired nursing home residents: perceptions of family members and certified nursing assistants. AB - Pain in cognitively impaired nursing home (NH) elders is difficult to detect. We report the results of the qualitative interview portion of a larger study that characterized the pain experience of cognitively impaired NH residents. Interviews were conducted with 16 family members or friends and 11 certified nursing assistants (CNAs) of 20 cognitively impaired NH residents experiencing pain. Analysis of the interviews yielded themes in family and CNA perceptions of pain in cognitively impaired NH residents. Family members and CNAs concurred that knowing the resident's usual behavior and daily habits was essential to being able to detect pain in cognitively impaired NH residents. Although a majority of family members reported that their relative was "stoic" about pain expression prior to NH placement, personal care information and historical information were perceived as helpful by nursing staff members. CNAs used facial and eye cues to detect pain and pain relief in their assigned residents. In addition, CNAs reported specific pain management strategies for their residents. Major themes related to pain detection in cognitively impaired NH residents identified in this investigation include: (a) knowing the resident; (b) importance of family input about previous pain behaviors in knowing the resident; (c) CNA reliance on face and eye cues for pain detection, particularly with residents who were nonverbal; and (d) the prevalence of pain with caregiving activities. Information from multiple sources can improve pain management strategies for cognitively impaired NH residents. PMID- 15359224 TI - Home treatment of pain for children and adolescents with sickle cell disease. AB - This ethnographic study examined family caregivers' perceptions of the effectiveness of comfort measures used at home for children with vaso-occlusive episodes (VOE) of sickle cell disease. Interviews were conducted in the hospital while children experienced VOE. Oral analgesics and most nonpharmacologic pain relief methods were rated by caregivers as only partly effective. Major themes that emerged from interview information reflected caregivers' efforts on behalf of their children in avoiding pain episodes, keeping a normal routine, "catching" the pain, getting their minds off the pain, helping the child get through the episode, and staying out of the hospital. Understanding the role of the family caregiver in home settings may assist health care professionals to provide more empathetic care to children with VOE and to their families when children are admitted to the hospital. Findings may encourage professionals to design the most effective methods of home pain management for these families. PMID- 15359227 TI - Aniridia and optic nerve hypoplasia. AB - BACKGROUND: This study evaluates, in patients with aniridia, the prevalence of optic nerve hypoplasia and its association with foveal hypoplasia. METHODS: The medical records of 56 patients with aniridia (31 female, 25 male, mean age 33 years, range 2-74 years) were retrospectively evaluated for optic nerve and foveal hypoplasia. The difference in prevalence of foveal hypoplasia in patients with and without optic nerve hypoplasia was compared using Fisher's exact test. RESULTS: Six of 56 patients, 10.7% (95% CI: 4.8-21.5%), had optic nerve hypoplasia; hypoplasia was found in both eyes of five binocular patients and in one monocular patient. The prevalence of foveal hypoplasia was higher in aniridia patients with optic nerve hypoplasia than in those without (50.0 vs 6.0%); this difference did not achieve statistical significance (P=0.10). CONCLUSIONS: Clinically apparent optic nerve hypoplasia is found in roughly 10% of patients with aniridia and may occur independently or in association with foveal hypoplasia. PMID- 15359228 TI - Long-term results of the management of silicone oil-induced raised intraocular pressure by diode laser cycloablation. AB - AIMS/BACKGROUND: To analyse the effectiveness and safety of cyclodiode treatment in eyes with silicone oil-related raised intraocular pressure (IOP) and to correlate the results with clinical features of treated eyes. METHODS: Retrospective review of case notes of all eyes that underwent cyclodiode treatment following injection of silicone oil at Moorfields Eye Hospital between April 1993 and January 2003 and were followed up for at least 1 year. RESULTS: In all, 38 patients were followed up for between 13 and 113 months. Totally, 17 had silicone oil in situ at the time of first treatment. The mean pretreatment IOP was 31.4 mmHg (SD 10.9), reducing to 18.6 mmHg (SD 8.6) at 1 year and to 13.9 mmHg (SD 8.4) at the final follow-up visit (P<0.001). Before treatment, 28 (73.7%) patients were on two or more topical medications. This was reduced to 18 (47.4%) patients (P=0.013) at 1 year and 13 (34.2%) patients (P=0.0007) at final follow-up. Use of oral acetazolamide for glaucoma was reduced from 16 (42.1%) patients precyclodiode to five (13.2%) patients at 1 year (P=0.0034) and three (7.9%) patients at the final visit (P=0.001). Four patients (10.5%) at 1 year and five patients (13.2%) at the final visit had hypotony (defined as IOP of less than 5 mmHg). One patient had enucleation 75 months following first cyclodiode treatment. CONCLUSION: Diode laser photocoagulation can successfully control silicone oil-induced raised intraocular pressure where medical treatment fails. Reduction of IOP appears to be maintained long term. PMID- 15359229 TI - Modification of Flieringa ring fixation. PMID- 15359230 TI - Immediate argon laser peripheral iridoplasty (ALPI) as initial treatment for acute phacomorphic angle-closure (phacomorphic glaucoma) before cataract extraction: a preliminary study. AB - PURPOSE: To study the safety and efficacy of immediate argon laser peripheral iridoplasty (ALPI) as initial treatment of acute phacomorphic angle-closure (phacomorphic glaucoma) before cataract extraction. METHODS: In all, 10 consecutive patients with acute phacomorphic angle-closure and intraocular pressure (IOP) > or =40 mmHg were recruited into the study. Each patient received topical atropine (1%) and timolol (0.5%), and immediate ALPI as initial treatment. The IOP at 15, 30, 60, and 120 min, and then 1 day, after ALPI were documented by applanation tonometry. Systemic IOP-lowering drugs were only started if IOP remained above 40 mmHg at 2 h after ALPI. Cataract extraction was subsequently performed as definitive treatment. RESULTS: In total, 10 patients (five male, five female), with a mean age+/-SD of 73.1+/-10.3 years were recruited. Mean duration of symptomatic attack was 128+/-232 h. After ALPI, the mean IOP was reduced from 56.1+/-12.5 to 45.3+/-14.5 mmHg at 15 min, 37.6+/-7.5 mmHg at 30 min, 34.2+/-9.7 mmHg at 60 min, 25.5+/-8.7 mmHg at 120 min, and 13.6+/ 4.2 mmHg at 1 day. In one patient, systemic acetazolamide was administered, because the IOP remained above 40 mmHg at 2 h after ALPI. All 10 patients had uncomplicated cataract extraction performed within 4 days after ALPI. No complications from the laser procedure were encountered. CONCLUSION: Immediate ALPI, replacing systemic antiglaucomatous medications, appeared to be safe and effective as first-line treatment of acute phacomorphic angle-closure. PMID- 15359231 TI - Light-near dissociation of pupil reactions as a presenting feature of von Hippel Lindau disease. PMID- 15359232 TI - Reply: limbal stem cell deficiency: a clinical chamaelon. PMID- 15359233 TI - Limbal stem cell deficiency: a clinical chameleon. PMID- 15359235 TI - Optic disc drusen associated with neovascularization of optic disc. PMID- 15359236 TI - Choroidal neovascularization (CNV) secondary to septic emboli from endocarditis: a case report. PMID- 15359237 TI - Choroidal folds after 25 gauge transconjunctival sutureless vitrectomy. PMID- 15359238 TI - Incidence and severity of iris pigmentation on latanoprost-treated glaucoma eyes. AB - PURPOSE: The purpose of the present study was to investigate the incidence and severity of iridial pigmentation under latanoprost topical use on brown eyes in Taiwan. METHODS: Retrospective review study was conducted from April 1999 to October 2001 in the Department of Ophthalmology, Taipei Veterans General Hospital, Taiwan, for glaucoma clinic monthly follow-up patients; 140 open-angle glaucoma patients on 0.005% latanoprost were enrolled. Analyses of iridial pigmentation incidence, grading, patient age distribution, side effect, and time course were performed. Boys-Smith pigment gradation lens was used as standard for semiquantitative iris pigmentation grading. RESULT: Before 0.005% latanoprost use, 90% of the patients enrolled were noted with iridial pigmentation grade I, and 10% were with grade I-II, but not reaching grade II scale standard. A total of 60 patients on 0.005% latanoprost developed increased pigmentation of the iris during the follow-up period. An increase of iris pigmentation was noted after an average of 7.27 months use of latanoprost (range 1-19 months, SD 2.65 months). For iridial pigmentation grading, 57.1, 30.7, 10.0, and 2.1% of our patients were noted to have grade I, II, III, and IV respectively. Most patients with latanoprost-induced iris hyperpigmentation were with grade II iridial pigmentation. There were 15 patients (10.7%) (10 female and five male) with hypertrichosis in the study group who were not compatible with the iridial pigmentation status. Among these patients, female patients had higher incidence of hypertrichosis than males, but this did not bother them. Only four patients (2.8%) were with conjunctiva chemosis and three patients (2.1%) with lid margin hyperpigmentation. CONCLUSION: Contrary to the belief that latanoprost rarely caused iris hyperpigmentation in yellow-brown eyes, our study showed that 42.8% iris hyperpigmentation did occur, especially after continual use for around 7 months. Higher hyperpigmentation incidence were noted in male than in female patients. This might be due to stronger adrenergic incidence in male than in female patients. Although hypertrichosis and increasing eyelid pigmentation together with iridial pigmentation represented a potentially permanent cosmetic side effect, they are very rare and occurred in no more than 3% in our patients. It is a good way to take Boys-Smith pigment gradation lens for iridial pigmentation grading and for long-term continual evaluation. The doctors should exert great care in differentiating drug-induced iris pigmentation and iris nevi from early stage uveal melanoma. PMID- 15359239 TI - Total hyphema following postoperative enoxaparin (Clexane). PMID- 15359240 TI - Valsalva and Purtscher's retinopathy with optic neuropathy in compressive thoracic injury. PMID- 15359241 TI - Treatment of polypoidal choroidal vasculopathy with transpupillary thermotherapy: an interventional case report. PMID- 15359242 TI - Dyslipidaemia and microvascular disease in the retina. AB - PURPOSE: There are few data on the effect of serum lipids on microvascular disease. This study assessed the relationships between serum lipid levels and microvascular disease, as seen in the retina, among participants who attended a population-based study in Australia (n=3654, aged 49+years). METHODS: Diameters of retinal arterioles and venules were measured from digitised photographs of each participant to obtain an estimate of generalised arteriolar narrowing. Focal arteriolar narrowing, arteriovenous nicking, and retinopathy lesions (microaneurysms, haemorrhages, hard/soft exudates) were graded using a standard protocol. Fasting blood tests were performed in 89% of subjects. Adjusted means were calculated using general linear models. Logistic regression models were used to determine the odds ratios for retinal microvascular signs. RESULTS: After controlling for age, sex, body mass index, smoking, and mean arterial blood pressure, elevated high-density lipoprotein cholesterol was associated with narrower retinal arterioles (Ptrend=0.002) and venules (Ptrend=0.03) and with increased odds of generalised arteriolar narrowing (odds ratio 1.6, 95% confidence interval 1.1-2.2 for the highest vs the lowest quintile of high density lipoprotein cholesterol). Serum triglyceride had a U-shaped relationship with venular diameter (Ptrend=0.003). We found no consistent pattern of association between serum total cholesterol or low-density lipoprotein cholesterol and any retinal microvascular signs. CONCLUSIONS: These findings suggest that microvascular disease in the retina may result from processes distinct from dyslipidaemia. PMID- 15359244 TI - Helicoidal peripapillary degeneration. PMID- 15359243 TI - Topically applied antibody fragments penetrate into the back of the rabbit eye. AB - AIMS: Antibody fragments have been shown to penetrate into the anterior chamber when applied to the cornea. The aim of this study was to investigate whether such fragments could penetrate into the vitreous cavity after topical administration to the ocular surface of rabbits. METHODS: An engineered single-chain variable domain antibody fragment with specificity for an irrelevant rat determinant was applied as a 50 microl eye drop to the eyes of live rabbits at 20-min intervals over 12 h. Eye drops contained 0.8-1.1 mg/ml protein in a buffered salt solution supplemented with penetration and viscosity enhancers. Samples were collected by paracentesis from the vitreous cavity immediately postmortem. Antibody fragments in these samples were quantified by measuring the binding activity to specific antigen, using flow cytometry. RESULTS: Topically applied antibody fragments were detectable in the vitreous of rabbit eyes after 4-12 h but had cleared at 12 h following the final eye drop. Concentrations of the antibody fragment in the vitreous samples were estimated to be 50-150 ng/ml at 12 h. Penetration of the parental whole antibody into the vitreous was not observed. CONCLUSION: Antibody fragments penetrate into the vitreous chamber of the rabbit eye after topical administration to the ocular surface. Such fragments may have therapeutic potential for diseases affecting the posterior segment. PMID- 15359245 TI - Two-port pars plana vitrectomy surgery: a prospective interventional case series. AB - BACKGROUND AND OBJECTIVE: Current literature review fails to disclose any series describing the use of two-port vitrectomy in adult patients. This study was performed to determine the feasibility and efficacy of pars plana vitrectomy surgery using two (rather than three or four)-port access for treatment of diabetic patients with nonclearing vitreous haemorrhage due to retinal neovascularization. DESIGN: Interventional prospective case series: to measure ability to allow for long-term resolution of chronic uncomplicated vitreous haemorrhage in diabetic patients, and to study the frequency and nature of complications associated with this technique. METHODS: Two-port pars plana vitrectomy (with endolaser treatment and membrane delamination if necessary) was performed in a prospective series of 12 consecutive diabetic patients with nonclearing vitreous haemorrhage due to retinal neovascularization. RESULTS: Successful removal of vitreous haemorrhage resulted in all patients. No visually significant intraoperative complications occurred. Best postoperative visual acuity correlated with lenticular and macular perfusion status. CONCLUSION: Two port pars plana vitrectomy is an efficient (and potentially safer and faster) alternative to the standard three-port vitrectomy in selected patients. PMID- 15359246 TI - Deep lamellar keratoplasty in corneal dermoid. PMID- 15359247 TI - Disposable devices for measuring intraocular pressure: a clinical study to assess their accuracy. AB - AIMS: We conducted a study to compare the accuracy of two commercially available disposable tonometry devices (the acrylic biprism, Tonosafe and the silicone shield, Tonoshield) with the standard Goldmann applanation tonometer (GAT). METHODS: A total of 80 eyes of 40 patients were randomly assigned to two groups. Group A (40 eyes) had the intraocular pressure IOP measured using GAT and the acrylic biprism (Tonosafe), while Group B (40 eyes) had IOP measured using GAT and the disposable silicone shield (Tonoshield). IOP was measured using the GAT both before and after the disposable device and average of the two standard Goldmann readings was compared with the IOP measured using the disposable devices. RESULTS: The mean difference in IOP in Group A was 0.29 mmHg (SD 0.54) (P=0.0018) while in Group B the mean difference in IOP was 2.09 mmHg (SD 1.23) (P<0.0001). The Altman-Bland method was used to assess the agreement between two methods of clinical measurement. This shows a positive bias of 0.29 (95% confidence interval (CI) 0.11-0.46) when using the disposable acrylic biprism and a positive bias of 2.09 (95% CI 1.69-2.48) with the silicone shield. CONCLUSION: The IOP recorded using the disposable acrylic biprism (Tonosafe) was in close agreement with the standard GAT. However, IOP readings obtained with the silicone shield (Tonoshield) tend to be higher than with the standard GAT. These facts should be kept in mind when recording IOP in clinical situations. PMID- 15359248 TI - Spontaneous improvement of serous maculopathy associated with congenital optic disc pit: an OCT study. PMID- 15359249 TI - A case of postoperative candida endophthalmitis. PMID- 15359250 TI - Tadalafil-associated anterior ischaemic optic neuropathy. PMID- 15359251 TI - Optical coherence tomography-assisted localization of retained intraocular foreign body. PMID- 15359252 TI - Metoprolol responding uveitis. PMID- 15359253 TI - Interventions for intermittent distance exotropia: review. AB - PURPOSE: Management decisions in intermittent distance exotropia vary and lack well-defined clinical guidelines. We undertook a systematic review in an attempt to clarify the effects of various surgical and nonsurgical treatments and to establish the significance of factors such as age with respect to outcome. The review was undertaken in collaboration with the Cochrane Eyes and Vision Group. METHODS: Electronic and manual searches were undertaken to identify randomised controlled trials of surgical or nonsurgical treatments for intermittent distance exotropia. We also contacted researchers active in this field for information about further published or unpublished studies. There were no language restrictions. Study abstracts identified from the searches were analysed independently by the two reviewers (SR and LG) and marked for inclusion, exclusion, or consideration. Reviewer analysis was compared and full papers for appropriate studies were requested. RESULTS: No randomised controlled trials were found that met our selection criteria. CONCLUSIONS: The current literature consists mainly of retrospective reviews. These are difficult to compare and analyse due to variations in definition, intervention criteria, and outcome measures. However, there appears to be an agreement that the nonsurgical treatment is more appropriate in small-angle deviations or as a supplement to surgery. Studies supporting both early and late surgical intervention were found, so the optimal timing of surgical intervention could not be concluded. There is a need for robust clinical trials to improve the evidence base for the management of this condition. PMID- 15359254 TI - Clinical features and surgical management of retinal detachment secondary to round retinal holes. AB - AIMS: The majority of rhegmatogenous retinal detachments result from pathological posterior vitreous detachment (PVD) and secondary horseshoe or giant retinal tears. Retinal detachment without PVD is usually associated with either retinal dialysis or round retinal holes. This study characterises the features, surgical outcome, and incidence of bilateral involvement of detachment associated with round retinal holes. METHODS: In all, 110 retinal detachments from 96 consecutive patients with retinal detachment secondary to round retinal holes were studied. Analysis of patient age, sex, refraction, preoperative visual acuity, presented symptoms, position and extent of detachment, number and distribution of holes present, posterior hyaloid membrane status, surgical management, outcome of surgery, and postoperative visual acuity were studied. RESULTS: The mean age for patients was 34 years with a marked female preponderance (64%) and myopia (83%). The posterior hyaloid membrane remained attached in 95 eyes (86%). In all, 45% patients had bilateral pathology, of which 33% had 'mirror image' distribution. Detachments were predominantly shallow (93%) and slow in progression (17%). A total of 100 detachments were repaired with cryotherapy and scleral buckling, eight with cryotherapy alone, and one with laser retinopexy. In all, 99% detachments were successfully reattached with a single procedure. The mean follow up period was 2 years. There were no instances of redetachment. CONCLUSIONS: Round hole detachments are slowly evolving detachments with attached vitreous gel in young, predominantly female myopes. Examination of the fellow eye should be mandatory as there is a high incidence of bilateral pathology. Scleral buckling procedures remained highly effective in this selected group of patients. PMID- 15359255 TI - Reversible night blindness in a patient with neuroendocrine tumour of pancreas. PMID- 15359256 TI - Beneficial effects of small-incision cataract surgery in patients demonstrating reduced ocular blood flow characteristics. AB - PURPOSE: To investigate intraocular pressure (IOP) and ocular haemodynamics following small incision cataract surgery. METHODS: Systemic and ocular haemodynamics, and IOP, were measured pre-operatively and 1 month post operatively, in 25 eyes of 25 patients (mean age 72.6+/-7.9 years) scheduled for small incision cataract surgery and intraocular lens implantation; these values were compared to an untreated age-matched healthy group (n=25, mean age 72.3+/ 5.0 years). For each eye, the Ocular Blood Flow Analyser (OBFA, Paradigm Inc., UT, USA) was used to obtain measurements of IOP, pulsatile ocular blood flow (POBF), pulse volume (PV), ocular pulse amplitude (PA), and pulse rate (PR). Systemic blood pressure measurements were obtained at each visit. Pre- and post operative values for the cataract group were compared using repeated measures analysis of variance, and compared to the normal group using analysis of variance. RESULTS: Pre-operatively, IOP was significantly higher in the cataract group compared to the age-matched normal control group (P<0.001). Cataract surgery resulted in a significant 16.7% reduction in IOP (P=0.001). Consequently, the post-operative IOP in the cataract group was similar to that of the normal group (P>0.05). Both pre- and post-operative POBF and PV values for the cataract group were significantly lower than for the normal age-matched group (P<0.0125). Small incision cataract surgery led to an 8.3% post-operative increase in POBF (P=0.0118) and 15.5% increase in PV (P<0.001). No changes in PA or PR were evident. CONCLUSION: These data suggest that cataract patients exhibit higher IOP and reduced ocular perfusion characteristics compared to an age-matched normal healthy group. Following cataract surgery, IOP reduced and ocular pulsatility improved. PMID- 15359257 TI - Intralenticular opacification of hydrophilic acrylic intraocular lenses. AB - PURPOSE: To find out the incidence of intraocular lens (IOL) optic opacification in a suspect IOL. METHODS: Retrospective review of patients' notes 3-4 years postimplant. RESULTS: In all, 56 IOLs showed signs of opacification, resulting in reduced vision out of 181 lenses traced. CONCLUSION: Certain hydrophilic acrylic IOLs like the model in question (SC60B-0UV) show opacification of the IOL, which may impair the vision of pseudophakic patients. However, not all lenses of this variety exhibit this cloudiness. Factors affecting this phenomenon are not yet completely clear but may include changes in the UV absorbant material incorporated in the lens substance itself. We report a series of patients who had received this lens and experienced loss of vision due to opacification of the IOL. In a follow-up ranging from 1 to 4 years postoperatively, a higher incidence of postoperative opacification of this lens emerges, after comparison with existing literature. IOL exchange in such patients is the only option. However, this mode of treatment should not be offered lightly as results may be guarded at best even in the face of a technically satisfactory procedure. PMID- 15359258 TI - Diabetic vitreopapillary traction and macular oedema. AB - PURPOSE: To describe an association between optic disc traction and diabetic macular oedema (DME) unresponsive to laser treatment. METHODS: A retrospective review of all patients with DME who attended our clinic between September 2001 and November 2003 was undertaken. The patients had undergone ophthalmic history and examination, fluorescein angiography, and optical coherence tomography (OCT) of the macular area and optic nerve head (ONH). A total of 10 nonvitrectomized eyes that were found to have an elevation of the ONH secondary to vitreopapillary traction were included in the analysis. Eyes with additional traction at the posterior pole were excluded. RESULTS: Out of the 10 eyes (seven patients, aged 47-79 years) with vitreo-papillary traction, nine had previously undergone argon laser photocoagulation(s) for DME. In seven eyes (seven patients), OCT verified the vitreopapillary traction as the sole traction, whereas in the fellow eyes of three patients vitreomacular traction was evident as well. In the seven eyes with only vitreopapillary traction, OCT demonstrated parapapillary serous retinal detachment in two eyes and a diffuse DME in all eyes (mean foveal thickness, 396+/-144 microm). Maximal thickness of the papillo-macular bundle site was adjoining the elevated ONH in three eyes, and was maximal at the central macula in the other four eyes. Ultrasonography (n=5) revealed an incomplete detachment of the posterior hyaloid in each, adherent only at the ONH. CONCLUSIONS: Diffuse DME unresponsive to laser treatment may be associated with vitreopapillary traction. Further studies should indicate whether these two phenomena could suggest a cause and effect in such eyes. PMID- 15359260 TI - Conjunctival necrosis and bleb leakage secondary to an adherent conjunctival foreign body. PMID- 15359261 TI - Retinal astrocytic hamartoma with exudation. PMID- 15359262 TI - Radial Descemet's membrane folds as a sign of pterygium traction. PMID- 15359264 TI - A late presentation of ocular quinine toxicity managed with a combination of vasodilatory treatments. PMID- 15359265 TI - A randomised, double-blind trial of topical ketorolac vs artificial tears for the treatment of episcleritis. AB - PURPOSE: To determine whether topical ketorolac (Acular) is more effective than artificial tears in treating the signs and symptoms of idiopathic episcleritis. METHODS: In this prospective, randomised, double-blind study, 38 eyes of 37 patients presenting with idiopathic episcleritis were allocated to receive either topical ketorolac (0.5%) or artificial tears three times a day for 3 weeks. The severity of patients' signs (episcleral injection and the number of clock hours affected) were recorded at weekly intervals. Patients' symptoms (perceived redness and pain scores) were recorded using a daily diary. RESULTS: There was no significant difference in the ophthalmic signs between the two groups at each assessment, including intensity of episcleral injection and the number of clock hours affected. No significant difference was found in the time to halve the baseline redness intensity scores (4.4 vs 6.1 days, P=0.2) or pain scores (3.6 vs 4.3 days, P=0.55). Significantly more patients on ketorolac reported stinging at the first follow-up visit (P<0.001). CONCLUSION: Topical ketorolac is not significantly better than artificial tears in treating the signs or symptoms of idiopathic episcleritis. PMID- 15359266 TI - Use of disposable prism tonometry in routine clinical practice. AB - AIM: To establish the reliability, efficacy, and safety of disposable prism tonometry and validate its routine use for screening as well as monitoring of glaucoma. METHODOLOGY: Intraocular pressure (IOP) of 400 eyes of 200 consecutive patients who attended the general ophthalmic clinic was checked with both the Goldmann applanation tonometer and a disposable tonometer prism by an experienced examiner after obtaining informed consent. The data were statistically analysed. RESULTS: The mean difference in the IOP between the two types of prisms was 0.1 mmHg (SD+/-1 mmHg). CONCLUSION: Disposable prism tonometry provides a reliable, effective, and safe alternative to Goldmann reusable prism tonometry for routine screening as well as monitoring of glaucoma with the advantages of eliminating the need for chemical disinfection and therefore eliminating the risk of crossinfection. PMID- 15359267 TI - Ocular and systemic side effects of latanoprost. PMID- 15359268 TI - Intravitreal triamcinolone improves vision in eyes with chronic diabetic macular oedema refractory to laser photocoagulation. AB - PURPOSE: To evaluate the clinical and visual outcome of an intravitreal injection of triamcinolone in patients with diabetic macular oedema refractory to laser treatment. MATERIAL AND METHODS: Prospective, interventional nonrandomised case series of 24 eyes of 24 patients that underwent intravitreal injection of 4 mg triamcinolone for chronic diabetic macular oedema refractory to previous laser treatment. All patients had best corrected visual acuity (bcva) on the LogMAR scale, near vision, slit-lamp biomicroscopy for the presence of clinically significant macular oedema, and intraocular pressure check at 1, 3, and 6 months. RESULTS: The average duration of macular oedema before the injection was 38.2 months (range=6-120 months) with the mean number of laser treatments before the injection being 2.2 (range=1-5). The visual acuity improved in 19 of the 23 (83%) eyes at 1 month, 16 of the 21 eyes (76%) at 3 months and six of 11 eyes (55%) at 6 months follow-up. The mean bcva improved by an average of 10, nine, and six letters, respectively, compared to the baseline at 1, 3, and 6 months (P=0.0002 at 1 month and 0.001 at 3 months with the Wilcoxon signed-ranked test). The intraocular pressure rose by an average of 2.4, 2.8, and 2.7 mmHg at the same follow-up visits. CONCLUSION: Intravitreal triamcinolone appears to be promising in the short term, for improving the vision in eyes with chronic diabetic macular oedema unresponsive to conventional laser treatment. Randomised controlled trials utilising varying doses of steroid are now required to define optimum treatment regimens. PMID- 15359269 TI - Photodynamic therapy for maculopathy due to radiation retinopathy. AB - BACKGROUND/AIMS: To describe the clinical outcomes of four eyes with macular oedema due to radiation retinopathy treated with verteporfin photodynamic therapy (PDT). METHODS: Interventional Case Series. Four charts of four patients who underwent PDT for macular oedema due to radiation retinopathy were reviewed. Snellen visual acuities, clinical examination and fundus photographs were performed before and after PDT. Main outcome measures were visual acuity, clinical examination before and after PDT. RESULTS: All four eyes had a marked reduction in hard exudates. Three of four eyes had an improvement in vision following the PDT. CONCLUSION: PDT may have a role in the treatment of macular oedema due to radiation retinopathy. PMID- 15359270 TI - Toll-like receptor 2-dependent bacterial sensing does not occur via peptidoglycan recognition. AB - Toll-like receptor 2 (TLR2) has been shown to recognize several classes of pathogen-associated molecular patterns including peptidoglycan (PG). However, studies linking PG with TLR2 recognition have relied mainly on the use of commercial Staphylococcus aureus PG and have not addressed TLR2 recognition of other PG types. Using highly purified PGs from eight bacteria (Escherichia coli, Pseudomonas aeruginosa, Yersinia pseudotuberculosis, Helicobacter pylori, Bacillus subtilis, Listeria monocytogenes, Streptococcus pneumoniae and S. aureus), we show that these PGs are not sensed through TLR2, TLR2/1 or TLR2/6. PG sensing is lost after removal of lipoproteins or lipoteichoic acids (LTAs) from Gram-negative and Gram-positive cell walls, respectively. Accordingly, purified LTAs are sensed synergistically through TLR2/1. Finally, we show that elicited peritoneal murine macrophages do not produce tumour necrosis factor-alpha or interleukin-6 in response to purified PGs, suggesting that PG detection is more likely to occur intracellularly (through Nod1/Nod2) rather than from the extracellular compartment. PMID- 15359271 TI - Enhanced bone formation in lipodystrophic PPARgamma(hyp/hyp) mice relocates haematopoiesis to the spleen. AB - The peroxisome proliferator-activated receptor gamma (PPARgamma) controls adipogenesis and metabolism. We demonstrate here that the absence of PPARgamma in fat has potent osteogenic activities, which affect haematopoiesis. The congenital absence of PPARgamma in fat of lipodystrophic PPARgamma(hyp/hyp) mice, strongly enhanced bone mass and consequentially reduced the bone-marrow cavity. Consistent with this, PPARgamma(hyp/hyp) mice had a significant decrease in bone marrow cellularity and resorted to extramedullary haematopoiesis in the spleen to maintain haematopoiesis. Our data indicate that antagonizing PPARgamma activity in fat could be an effective way to combat osteoporosis and suggest that haematopoietic function should be scrutinized in lipodystrophic subjects. PMID- 15359272 TI - DSS1 is required for RAD51 focus formation and genomic stability in mammalian cells. AB - BRCA2 is a breast cancer susceptibility gene implicated in the repair of double strand breaks by homologous recombination with RAD51. BRCA2 associates with a 70 amino-acid protein, DSS1, but the functional significance of this interaction has remained unclear. Recently, deficiency of a DSS1 orthologue in the fungus Ustilago maydis has been shown to cause a defect in recombinational DNA repair. Here we have investigated the consequences of DSS1 depletion in mammalian cells. We show that like BRCA2, DSS1 is required for DNA damage-induced RAD51 focus formation and for the maintenance of genomic stability, indicating a function conserved from lower eukaryotes to humans. However, DSS1 seems to be not required for BRCA2 or RAD51 stability or for BRCA2 and RAD51 to interact, raising the possibility that DSS1 may be required for the BRCA2-RAD51 complex to become associated with sites of DNA damage. PMID- 15359273 TI - Members of the SAGA and Mediator complexes are partners of the transcription elongation factor TFIIS. AB - TFIIS, an elongation factor encoded by DST1 in Saccharomyces cerevisiae, stimulates transcript cleavage in arrested RNA polymerase II. Two components of the RNA polymerase II machinery, Med13 (Srb9) and Spt8, were isolated as two hybrid partners of the conserved TFIIS N-terminal domain. They belong to the Cdk8 module of the Mediator and to a subform of the SAGA co-activator, respectively. Co-immunoprecipitation experiments showed that TFIIS can bind the Cdk8 module and SAGA in cell-free extracts. spt8Delta and dst1Delta mutants were sensitive to nucleotide-depleting drugs and epistatic to null mutants of the RNA polymerase II subunit Rpb9, suggesting that their elongation defects are mediated by Rpb9. rpb9Delta, spt8Delta and dst1Delta were lethal in cells lacking the Rpb4 subunit. The TFIIS N-terminal domain is also strictly required for viability in rpb4Delta, although it is not needed for binding to RNA polymerase II or for transcript cleavage. It is proposed that TFIIS and the Spt8-containing form of SAGA co operate to rescue RNA polymerase II from unproductive elongation complexes, and that the Cdk8 module temporarily blocks transcription during transcript cleavage. PMID- 15359274 TI - Reaction cycle of the yeast Isw2 chromatin remodeling complex. AB - Members of the ISWI family of chromatin remodeling factors hydrolyze ATP to reposition nucleosomes along DNA. Here we show that the yeast Isw2 complex interacts with DNA in a nucleotide-dependent manner at physiological ionic strength. Isw2 efficiently binds DNA in the absence of nucleotides and in the presence of a nonhydrolyzable ATP analog. Conversely, ADP promotes the dissociation of Isw2 from DNA. In contrast, Isw2 remains bound to mononucleosomes through multiple cycles of ATP hydrolysis. Solution studies show that Isw2 undergoes nucleotide-dependent alterations in conformation not requiring ATP hydrolysis. Our results indicate that during an Isw2 remodeling reaction, hydrolysis of successive ATP molecules coincides with cycles of DNA binding, release, and rebinding involving elements of Isw2 distinct from those interacting with nucleosomes. We propose that progression of the DNA-binding site occurs while nucleosome core contacts are maintained and generates a force dissipated by disruption of histone-DNA interactions. PMID- 15359275 TI - Bystander gene activation by a locus control region. AB - Random assortment of genes within mammalian genomes establishes the potential for interference between neighboring genes with distinct transcriptional specificities. Long-range transcriptional controls further increase this potential. Exploring this problem is of fundamental importance to understanding gene regulation. In the human genome, the Igbeta (CD79b) gene is situated between the pituitary-specific human growth hormone (hGH) gene and its locus control region (hGH LCR). Igbeta protein is considered B-cell specific; its only known role is in B-cell receptor signaling. Unexpectedly, we found that hIgbeta is transcribed at high levels in the pituitary. This Igbeta transcription is dependent on pituitary-specific epigenetic modifications generated by the hGH LCR. In contrast, expression of Igbeta at its native site in B cells is independent of hGH LCR activity. These studies demonstrated that a gene with tissue-restricted transcriptional determinants (B cell) can be robustly activated in an unrelated tissue (pituitary) due to fortuitous positioning within an active chromatin domain. This 'bystander' gene activation pathway impacts on current concepts of tissue specificity and models of active chromatin domains. PMID- 15359276 TI - Structure and DNA-binding properties of the cytolysin regulator CylR2 from Enterococcus faecalis. AB - Enterococcus faecalis is one of the major causes for hospital-acquired antibiotic resistant infections. It produces an exotoxin, called cytolysin, which is lethal for a wide range of Gram-positive bacteria and is toxic to higher organisms. Recently, the regulation of the cytolysin operon was connected to autoinduction by a quorum-sensing mechanism involving the CylR1/CylR2 two-component regulatory system. We report here the crystal structure of CylR2 and its properties in solution as determined by heteronuclear NMR spectroscopy. The structure reveals a rigid dimer containing a helix-turn-helix DNA-binding motif as part of a five helix bundle that is extended by an antiparallel beta-sheet. We show that CylR2 is a DNA-binding protein that binds specifically to a 22 bp fragment of the cytolysin promoter region. NMR chemical shift perturbation experiments identify surfaces involved in DNA binding and are in agreement with a model for the CylR2/DNA complex that attributes binding specificity to a complex network of CylR2/DNA interactions. Our results propose a mechanism where repression is achieved by CylR2 obstruction of the promoter preventing biosynthesis of the cytolysin operon transcript. PMID- 15359277 TI - Two WXXF-based motifs in NECAPs define the specificity of accessory protein binding to AP-1 and AP-2. AB - The adaptor proteins AP-2 and AP-1/GGAs are essential components of clathrin coats at the plasma membrane and trans-Golgi network, respectively. The adaptors recruit accessory proteins to clathrin-coated pits, which is dependent on the adaptor ear domains engaging short peptide motifs in the accessory proteins. Here, we perform an extensive mutational analysis of a novel WXXF-based motif that functions to mediate the binding of an array of accessory proteins to the alpha-adaptin ear domain of AP-2. Using nuclear magnetic resonance and mutational studies, we identified WXXF-based motifs as major ligands for a site on the alpha ear previously shown to bind the DPW-bearing proteins epsin 1/2. We also defined the determinants that allow for specific binding of the alpha-ear motif to AP-2 as compared to those that allow a highly related WXXF-based motif to bind to the ear domains of AP-1/GGAs. Intriguingly, placement of acidic residues around the WXXF cores is critical for binding specificity. These studies provide a structural basis for the specific recruitment of accessory proteins to appropriate sites of clathrin-coated vesicle formation. PMID- 15359278 TI - Rad18 guides poleta to replication stalling sites through physical interaction and PCNA monoubiquitination. AB - The DNA replication machinery stalls at damaged sites on templates, but normally restarts by switching to a specialized DNA polymerase(s) that carries out translesion DNA synthesis (TLS). In human cells, DNA polymerase eta (poleta) accumulates at stalling sites as nuclear foci, and is involved in ultraviolet (UV)-induced TLS. Here we show that poleta does not form nuclear foci in RAD18(-/ ) cells after UV irradiation. Both Rad18 and Rad6 are required for poleta focus formation. In wild-type cells, UV irradiation induces relocalization of Rad18 in the nucleus, thereby stimulating colocalization with proliferating cell nuclear antigen (PCNA), and Rad18/Rad6-dependent PCNA monoubiquitination. Purified Rad18 and Rad6B monoubiquitinate PCNA in vitro. Rad18 associates with poleta constitutively through domains on their C-terminal regions, and this complex accumulates at the foci after UV irradiation. Furthermore, poleta interacts preferentially with monoubiquitinated PCNA, but poldelta does not. These results suggest that Rad18 is crucial for recruitment of poleta to the damaged site through protein-protein interaction and PCNA monoubiquitination. PMID- 15359279 TI - Structural determinants of phosphoinositide selectivity in splice variants of Grp1 family PH domains. AB - The pleckstrin homology (PH) domains of the homologous proteins Grp1 (general receptor for phosphoinositides), ARNO (Arf nucleotide binding site opener), and Cytohesin-1 bind phosphatidylinositol (PtdIns) 3,4,5-trisphosphate with unusually high selectivity. Remarkably, splice variants that differ only by the insertion of a single glycine residue in the beta1/beta2 loop exhibit dual specificity for PtdIns(3,4,5)P(3) and PtdIns(4,5)P(2). The structural basis for this dramatic specificity switch is not apparent from the known modes of phosphoinositide recognition. Here, we report crystal structures for dual specificity variants of the Grp1 and ARNO PH domains in either the unliganded form or in complex with the head groups of PtdIns(4,5)P(2) and PtdIns(3,4,5)P(3). Loss of contacts with the beta1/beta2 loop with no significant change in head group orientation accounts for the significant decrease in PtdIns(3,4,5)P(3) affinity observed for the dual specificity variants. Conversely, a small increase rather than decrease in affinity for PtdIns(4,5)P(2) is explained by a novel binding mode, in which the glycine insertion alleviates unfavorable interactions with the beta1/beta2 loop. These observations are supported by a systematic mutational analysis of the determinants of phosphoinositide recognition. PMID- 15359280 TI - Essential role of Mia40 in import and assembly of mitochondrial intermembrane space proteins. AB - Mitochondria import nuclear-encoded precursor proteins to four different subcompartments. Specific import machineries have been identified that direct the precursor proteins to the mitochondrial outer membrane, inner membrane or matrix, respectively. However, a machinery dedicated to the import of mitochondrial intermembrane space (IMS) proteins has not been found so far. We have identified the essential IMS protein Mia40 (encoded by the Saccharomyces cerevisiae open reading frame YKL195w). Mitochondria with a mutant form of Mia40 are selectively inhibited in the import of several small IMS proteins, including the essential proteins Tim9 and Tim10. The import of proteins to the other mitochondrial subcompartments does not depend on functional Mia40. The binding of small Tim proteins to Mia40 is crucial for their transport across the outer membrane and represents an initial step in their assembly into IMS complexes. We conclude that Mia40 is a central component of the protein import and assembly machinery of the mitochondrial IMS. PMID- 15359281 TI - Centrosome amplification induced by DNA damage occurs during a prolonged G2 phase and involves ATM. AB - Centrosomes are the principal microtubule organising centres in somatic cells. Abnormal centrosome number is common in tumours and occurs after gamma irradiation and in cells with mutations in DNA repair genes. To investigate how DNA damage causes centrosome amplification, we examined cells that conditionally lack the Rad51 recombinase and thereby incur high levels of spontaneous DNA damage. Rad51-deficient cells arrested in G2 phase and formed supernumerary functional centrosomes, as assessed by light and serial section electron microscopy. This centrosome amplification occurred without an additional DNA replication round and was not the result of cytokinesis failure. G2-to-M checkpoint over-ride by caffeine or wortmannin treatment strongly reduced DNA damage-induced centrosome amplification. Radiation-induced centrosome amplification was potentiated by Rad54 disruption. Gene targeting of ATM reduced, but did not abrogate, centrosome amplification induced by DNA damage in both the Rad51 and Rad54 knockout models, demonstrating ATM-dependent and -independent components of DNA damage-inducible G2-phase centrosome amplification. Our data suggest DNA damage-induced centrosome amplification as a mechanism for ensuring death of cells that evade the DNA damage or spindle assembly checkpoints. PMID- 15359282 TI - Role of the fission yeast SUMO E3 ligase Pli1p in centromere and telomere maintenance. AB - Sumoylation represents a conserved mechanism of post-translational protein modification. We report that Pli1p, the unique fission yeast member of the SP RING family, is a SUMO E3 ligase in vivo and in vitro. pli1Delta cells display no obvious mitotic growth defects, but are sensitive to the microtubule destabilizing drug TBZ and exhibit enhanced minichromosome loss. The weakened centromeric function of pli1Delta cells may be related to the defective heterochromatin structure at the central core, as shown by the reduced silencing of an ura4 variegation reporter gene inserted at cnt and imr. Interestingly, pli1Delta cells also exhibit enhanced loss of the ura4 reporter at these loci, likely by gene conversion using homologous sequences as information donors. Moreover, pli1Delta cells exhibit consistent telomere length increase, possibly achieved by a similar process. Point mutations within the RING finger of Pli1p totally or partially reproduce the pli1 deletion phenotypes, thus correlating with their sumoylation activity. Altogether, these results strongly suggest that Pli1p, and by extension sumoylation, is involved in mechanisms that regulate recombination in particular heterochromatic repeated sequences. PMID- 15359283 TI - Requirement of IS911 replication before integration defines a new bacterial transposition pathway. AB - Movement of transposable elements is often accompanied by replication to ensure their proliferation. Replication is associated with both major classes of transposition mechanisms: cut-and-paste and cointegrate formation (paste-and copy). Cut-and-paste transposition is often activated by replication of the transposon, while in cointegrate formation replication completes integration. We describe a novel transposition mechanism used by insertion sequence IS911, which we call copy-and-paste. IS911 transposes using a circular intermediate (circle), which then integrates into a target. We demonstrate that this is derived from a branched intermediate (figure-eight) in which both ends are joined by a single strand bridge after a first-strand transfer. In vivo labelling experiments show that the process of circle formation is replicative. The results indicate that the replication pathway not only produces circles from figure-eight but also regenerates the transposon donor plasmid. To confirm the replicative mechanism, we have also used the Escherichia coli terminators (terC) which, when bound by the Tus protein, inhibit replication forks in a polarised manner. Finally, we demonstrate that the primase DnaG is essential, implicating a host-specific replication pathway. PMID- 15359284 TI - The novel E3 ubiquitin ligase Tiul1 associates with TGIF to target Smad2 for degradation. AB - Ubiquitin-dependent degradation plays an important role in the negative regulation of TGF-beta signaling. Here, we identify Tiul1 (for TGIF interacting ubiquitin ligase 1), a novel E3 ubiquitin ligase that inhibits TGF-beta signaling by targeting both the activated receptor and Smad2 for degradation. Tiul1 associates constitutively with Smad7 and induces degradation of the activated type I receptor without affecting the expression levels of Smad7. Tiul1 can also interact with Smad2 and the nuclear corepressor TGIF upon activation of TGF-beta signaling. Like Smad7, the steady-state levels of TGIF are not affected by Tiul1, but the interaction of Tiul1 with TGIF allows this ubiquitin ligase to target Smad2 for degradation. Consistent with this, overexpression of Tiul1 suppressed TGF-beta-induced growth arrest and transcriptional responses. In addition, silencing of Tiul1 or TGIF genes by siRNA resulted in suppression of the TGF-beta dependent degradation of Smad2 and an enhancement of TGF-beta-mediated gene expression. These results reveal a new role for TGIF as a component of a ubiquitin ligase complex that mediates the degradation of Smad2 in response to TGF-beta signaling. PMID- 15359285 TI - Protection of hematopoietic stem cells from pemetrexed toxicity by retroviral gene transfer with a mutant dihydrofolate reductase-mutant thymidylate synthase fusion gene. AB - Myelosuppression is one of the major side effects of most anticancer drugs. To confer myeloprotection, our laboratory generated drug-resistant mutants of select target human enzymes for gene transfer to the bone marrow. Mutants of two of these enzymes, dihydrofolate reductase (DHFR F/S) and thymidylate synthase (TS G52S), were previously shown to confer resistance to methotrexate and 5-FU, respectively, and recently a fusion cDNA of both mutant enzymes (DHFR F/S-TS G52S) was shown to confer dual resistance to both antimetabolites. In this study, we examined the sensitivity of the DHFR F/S-TS G52S fusion protein to the multitargeted antifolate, pemetrexed (LY231514, Alimta), which targets both DHFR and TS and is currently in phase III trials for the treatment of solid tumors and in combination with cisplatin has been shown to be an advance in the treatment of mesothelioma. The K(i) for the DHFR F/S portion of the purified fusion protein to pemetrexed was increased by greater than 9000-fold when compared to wtDHFR (8000 versus 0.86 nM), while the K(i) for the TS G52S portion of the fusion protein to pemetrexed was similar to that of wtTS (2.8 versus 3.1 nM). When the fusion gene was retrovirally transduced into NIH 3T3 fibroblasts, the IC(50) to pemetrexed was three- to four-fold higher than cells transduced with DHFR F/S or TS G52S alone (163 versus 53 and 45 nM, respectively). Similarly, expression of the DHFR F/S-TS G52S fusion gene in retrovirally transduced mouse marrow cells resulted in an increased survival of CFU-GM colonies when compared to cells transduced with either of the mutants alone. Co-expression of mutant DHFR and TS enzymes has additive effects in conferring resistance to pemetrexed-induced toxicity. This construct may be useful for conferring myeloprotection to patients receiving this drug. PMID- 15359286 TI - Cancer-specific activation of the survivin promoter and its potential use in gene therapy. AB - Survivin is expressed in many cancers but not in normal adult tissues and is transcriptionally regulated. To test the feasibility of using the survivin promoter to induce cancer-specific transgene expression in lung cancer gene therapy, a vector expressing a luciferase gene driven by the survivin promoter was constructed and evaluated in vitro and in vivo. We found that the survivin promoter was generally more highly activated in cancer cell lines than in normal and immortalized normal cell lines. When delivered intravenously by DNA:liposome complexes, the survivin promoter was more than 200 times more cancer specific than the cytomegalovirus promoter in vivo. To identify lung cancer patients who may benefit from gene therapy with the survivin promoter, we measured survivin protein expression in surgical specimens of 75 non-small-cell lung cancers and 10 normal lung tissues by immunohistochemical staining and found that survivin is expressed in most of the non-small-cell lung cancers tested (81%, 61 of 75) but none of the normal lung tissues. The survivin promoter also induced transgene expression of a mutant Bik in cancer cells, which suppressed the growth of cancer cells in vitro and in vivo. These results indicate that the survivin promoter is a cancer-specific promoter for various cancers and that it may be useful in cancer gene therapy. PMID- 15359287 TI - Antiangiogenic cancer gene therapy by adeno-associated virus 2-mediated stable expression of the soluble FMS-like tyrosine kinase-1 receptor. AB - Antiangiogenic gene transfer has the potential to be more efficacious than protein-based therapies or pharmacotherapies for the control of solid tumor growth, invasion and metastasis. For a sustained antiangiogenic effect, a vector capable of long-term expression without vector-associated immunity or toxicity is advantageous. The present study evaluated the potential of a recombinant adeno associated virus-2 (rAAV) encoding the human soluble FMS-like tyrosine kinase receptor 1 (sFlt-1), which functions by both sequestering vascular endothelial growth factor (VEGF) and forming inactive heterodimers with other membrane spanning VEGF receptors, in vitro and in vivo. Results indicated significant growth inhibitory activity of the transgenic factor in a human umbilical vein endothelial cell proliferation assay in vitro and protection against the growth of an angiogenesis-dependent human ovarian cancer cell line, SKOV3.ip1, xenograft in vivo with increased disease-free survival. Stable expression of the secretory factor and transgene persistence were confirmed by immunohistochemistry and in situ hybridization analyses, respectively. Increased therapeutic effects on both the growth index of the implanted tumor cells and tumor-free survival also correlated with an increasing dose of the vector used. These studies indicate that rAAV-mediated sFlt-1 gene therapy may be a feasible approach for inhibiting tumor angiogenesis, particularly as an adjuvant/therapy. PMID- 15359288 TI - Molecular mechanisms regulating the tumor-targeting potential of splice-activated gene expression. AB - Previous studies have suggested that differences in the ability of normal and malignant cells to process certain alternatively spliced pre-mRNA transcripts can be exploited as a potentially powerful means of targeting the expression of therapeutic genes to tumor cells in vivo and in vitro. Specifically, it was shown that efficient processing of minigene constructs containing the alternatively spliced CD44 exons v9 and v10 only occurs in tumor cells that express CD44 isoforms that incorporate these exons (e.g. CD44R1). In the present study, efforts were made to define the molecular mechanisms that underlie the apparent specificity of this process. RT-PCR analysis and DNA sequencing were used to characterize the various splicing events that occur between CD44 exons v8, v9 and v10 following transfection of minigene constructs containing these various exons into CD44R1-positive (PC3) and CD44R1-negative (T24) cell lines. The results obtained confirm that although the v8-v9 intron is efficiently removed in both CD44R1-positive and CD44R1-negative cells, the corresponding v9-v10 intron is accurately spliced and the exons appropriately joined only in lines that express v10-containing CD44 isoforms (e.g. PC3). In CD44R1-negative cell lines (e.g. T24) alternative 5' and 3' splice sites located within the v9-v10 intron are preferentially used, resulting in various portions of the intron being retained within the final processed mRNA product. It is proposed that identification of these functionally important intronic sequence elements will facilitate the development of second generation "splice activated gene expression" vectors that may prove useful in various cancer gene therapy applications. PMID- 15359289 TI - Decreased tumorigenic potential of EphA2-overexpressing breast cancer cells following treatment with adenoviral vectors that express EphrinA1. AB - The EphA2 receptor tyrosine kinase is frequently overexpressed in invasive breast cancer cells. Moreover, these malignant cells have unstable cell-cell contacts, which preclude EphA2 from interacting with its ligand, EphrinA1, which is anchored to the membrane of adjacent cells. This defect is important because ligand binding causes EphA2 to transmit signals that negatively regulate tumor cell growth and survival, whereas the absence of ligand binding favors these same behaviors. In our present study, human adenoviral type 5 (HAd) vectors were engineered to express secreted-forms of EphrinA1. These vectors were used to infect MDA-MB-231 human breast cancer cells, or MCF-10A human breast epithelial cells providing matched controls. Infection with HAd-EphrinA1-Fc (HAd vector expressing extracellular domain of human EphrinA1 attached to Fc portion of human IgG1 heavy chain) caused increased EphA2 activation and turnover and consequently decreased tumor cell viability in soft agar assays. Consistent with this observation, infection of MDA-MB-231 cells with HAd-EphrinA1-Fc prevented tumor formation in xenograft models. Furthermore, therapeutic modeling via intratumoral inoculation revealed that HAd-EphrinA1-Fc significantly inhibited subsequent tumor growth as compared to matched controls. These results suggest that targeting of EphA2 with adenoviral vectors may have therapeutic value. PMID- 15359290 TI - Immune properties of recombinant vaccinia virus encoding CD154 (CD40L) are determined by expression of virally encoded CD40L and the presence of CD40L protein in viral particles. AB - Expression of costimulatory molecules by recombinant poxviruses is a promising strategy for enhancing therapeutic vaccines. CD40-CD40L interactions are critical for conditioning dendritic cells (DC) and priming T- and B-cell immunity. We constructed a vaccinia virus expressing murine CD40L (rV-CD40L) and studied its immunomodulatory properties in vitro. Direct DC infection with control vaccinia or psoralen/UV-inactivated rV-CD40L stimulated high levels of interleukin 12 (IL 12) release. However, replication-competent rV-CD40L did not stimulate IL-12 under similar conditions. We observed a high level of CD40L protein on purified viral particles and demonstrated that induction of IL-12 by nonreplicating rV CD40L was blocked by anti-CD40 antibodies suggesting that functional CD40L on viral particles contributed to alterations in IL-12 synthesis. Since cross presentation of tumor-associated antigens by DC is augmented by viral infection of tumor cells, we infected MC38 murine colon carcinoma cells with rV-CD40L. Infected cells stimulated IL-12 secretion by DC and proliferation of B cells and DX5(+) (NK/NKT) cells through direct CD40-CD40L interaction. A subpopulation of NKT cells expressing CD40 (NK1.1(+), CD3(lo)) appeared to be a major effector population responding to MC38/rV-CD40L. These results highlight the complex immune regulatory effects of rV-CD40L defined by the cumulative effects of CD40L expression, presence of CD40L protein in viral particles, and the replication potential of the virus. PMID- 15359291 TI - Experimental infections of humans with wild-type adenoviruses and with replication-competent adenovirus vectors: replication, safety, and transmission. AB - Replication-competent (RC) adenoviruses (Ads) are increasingly being developed as oncolytic vectors and as vehicles for delivering vaccine antigens. Although the safety of such vectors in humans is of paramount importance, these vectors pose additional special concerns. Specifically, the prospect of causing Ad-mediated disease in the patient, the amount and sites of Ad replication, the possibility of virus shedding leading to unintended transmission to patient contacts, and the potential for persistence in the inoculated individual must be evaluated. Previous experience with administration of wild-type and RC recombinant Ads to humans may shed light on some of these issues. Experimental infections of humans with natural Ad isolates and RC recombinant vectors show that in adults Ads cause mild or no disease, particularly with Ad serotypes 2 and 5, the serotypes most often used to make recombinant constructs. Other studies show that Ad can replicate in experimentally infected persons, that in some situations Ads can be shed and transmitted to close contacts, and that there is evidence for persistent/latent Ad infection in naturally infected individuals. Overall, these studies indicate that Ads can be safely administered to humans for the treatment of cancer and as antigen delivery vehicles suggesting that the continued development of RC oncolytic and vaccine vectors should be pursued. PMID- 15359292 TI - Herpes simplex virus thymidine kinase gene transduction enhances tumor growth rate and cyclooxygenase-2 expression in murine colon cancer cells. AB - Transduction of tumor cells with herpes simplex virus thymidine kinase (HSV-tk) gene and subsequent treatment with the prodrug ganciclovir (GCV) is the most common system utilized to date for "suicide" gene therapy of cancer. In the current report, we show that HSV-tk gene transduction enhances tumor growth rate of murine colon cancer cells, that are implanted subcutaneously in syngeneic mice, and enhances cyclooxygenase-2 (COX-2) protein expression and prostaglandin E(2) (PGE(2)) release in vitro and in vivo. It is further shown that the observed phenomenon is related to the presence of the HSV-tk sequence insert in the retroviral vector used for HSV-tk gene delivery. Transduction of murine colon cancer cells with control vector, carrying the neomycin-resistance gene alone, failed to increase tumor growth rate and COX-2 protein expression or PGE(2) production. On the contrary, it even decreased tumor growth, COX-2 protein expression and PGE(2.) The growth rate of HSV-tk-transduced murine tumors was significantly reduced by treatment with the selective COX-2 inhibitor nimesulide. Additionally, we demonstrate herein that both enhanced growth rate of HSV-tk transduced murine tumors and increased levels of PGE(2) in HSV-tk-transduced cells persist upon the development of GCV resistance. Taken together, these results provide a better understanding of the direct effect of HSV-tk gene transduction on tumor cell biology and target tumor development. PMID- 15359293 TI - Microglia rules: insights into microglial-neuronal signaling. PMID- 15359294 TI - The changing face of dental education. PMID- 15359295 TI - Help at hand. PMID- 15359296 TI - Student self-assessment of essential skills in dental surgery. AB - OBJECTIVE: To assess whether student self-assessments of essential skills in dental surgery are reliable indicators of the strengths and weaknesses of the clinical aspects of the dental school curriculum. This was done as part of an ongoing evaluation of the curriculum of the Faculty of Dental Sciences of the University of Peradeniya in Sri Lanka. DESIGN: Cross-sectional surveys of 5 different classes of students using a pre-tested questionnaire. SETTING: The Faculty of Dental Sciences of the University of Peradeniya, Sri Lanka, which is the only dental school in the island. MATERIALS & METHODS: A questionnaire was developed specifically for the purpose of eliciting student self-confidence in 46 essential skills on a three point scale. It was administered to 5 different classes of students, immediately or soon after their respective Final BDS examinations. The 5 classes had followed similar curricular formats. The surveys were conducted on a voluntary and anonymous basis. RESULTS: Results show a remarkable consistency in the self-ratings done by the five different classes of students who followed the same curriculum during five different time periods. Students were least confident in the skills related to the management of medical emergencies and of oral manifestations of systemic diseases while they were most confident in skills related to the care of periodontal disease and caries. CONCLUSION: Student self-assessments of skills is a useful tool for evaluating the clinical training provided in the dental school curriculum. Students were most confident in managing problems that they most frequently encountered in the dental school. The areas in which they were least confident require curricular reform. PMID- 15359297 TI - "On Track"--an educational resource to support dental SHO training. AB - This paper describes the development, implementation and evaluation of an educational resource, "On Track", designed to support dental SHOs throughout their time in post, thereby helping them gain optimum benefit from this important stage in their early postgraduate training. The need for such a resource was highlighted by a previous study undertaken by the paper's key authors with SHOs and their trainers in a UK dental school. Among several areas of change identified by both trainers and trainees were the needs to develop the SHO experience into more than an apprenticeship by improving the trainer/trainee relationship, and to encourage trainee-centred learning and self-appraisal. The key aims of "On Track" were therefore to define the outcomes of SHO training, to facilitate the relationship between SHO and educational supervisor (trainer), and to encourage the SHO in reflective learning and self-appraisal."On Track" was subject to developmental testing with both trainers and trainees to ensure that it could achieve these aims and the resultant resource is described, along with its implementation and evaluation. "On Track" met with mixed success and the reasons behind this and conclusions drawn for the introduction of any future similar resources are outlined. PMID- 15359298 TI - Key skills for newly qualified dentists: an evaluation of a West Midlands initiative. AB - INTRODUCTION: This paper reports an evaluation of the West Midlands Key Skills initiative that provides a framework for learning during vocational training (VT). METHOD: The 48 vocational dental practitioners (VDPs) who began their VT in August 2001 in the West Midlands were surveyed at the start and end of training (45 completed both surveys). They rated their confidence and experience in the 31 components of the Six Key Skills on visual analogue scales. Views were elicited in a survey of both VDPs (47 returns) and their general practice trainers (44 returns). Semi-structured interviews were also held with a stratified sample of 9 trainers and all four VT advisors. RESULTS: In terms of VDP progress, a statistically significant increase in confidence and experience was found in each of the 31 components. Six themes were identified in the views data. (i) Supporting the development of Key Skills; (ii) workload implications; (iii) the "right" six? (iv) links with the advanced diploma (MFGDP(UK)); (v) assessment of VT; and, (vi) consistency and quality assurance. CONCLUSION: VT successfully develops the confidence and experience of newly qualified dentists in the Six Key Skills and has been well received by the majority of VDPs, trainers and advisors in the West Midlands. PMID- 15359299 TI - Employing an overseas dentist. PMID- 15359300 TI - Take a note. PMID- 15359301 TI - Who's in control? PMID- 15359302 TI - Oral lichen planus. PMID- 15359303 TI - Antibiotic cover. PMID- 15359304 TI - An imperfect system? PMID- 15359305 TI - Patient confidentiality. PMID- 15359306 TI - Plain common sense. PMID- 15359307 TI - Risk management. PMID- 15359316 TI - Endodontics: Part 2 Diagnosis and treatment planning. AB - As with all dental treatment, a detailed treatment plan can only be drawn up when a correct and accurate diagnosis has been made. It is essential that a full medical, dental and demographic history be obtained, together with a thorough extra-oral and intra-oral examination. This part considers the classification of diseases of the dental pulp, together with various diagnostic aids to help in determining which condition is present, and the appropriate therapy. PMID- 15359317 TI - Phantom bite revisited. AB - The term 'phantom bite' is used to describe an uncommon condition in which patients are preoccupied with their dental occlusion, believing that it is abnormal. The condition is remarkable for the nature of the involved explanations and interpretations that the patients give and for their persistence in trying to find a solution to what appears to be a relatively minor problem. Three clinical cases that illustrate the nature of this condition are presented and problems associated with the management of affected patients are discussed. Phantom bite can be a disabling disorder which is difficult to treat. Available evidence suggests that the symptoms cannot be improved by occlusal treatments. It is therefore essential to avoid extensive irreversible restorative treatment. General dental practitioners should refer patients for specialist opinion and management. Psychiatric assessment with recommendations for management should be obtained if possible. The prognosis is poor for symptom elimination but need not necessarily be poor for patients' overall functioning and well-being. It is suggested that emphasis should be placed on building adaptive coping skills. Further research is needed to elucidate the nature of the condition to improve treatment. PMID- 15359324 TI - A prospective study to investigate the relationship between periodontal disease and adverse pregnancy outcome. AB - OBJECTIVE: This study aimed to investigate a relationship between maternal periodontal disease and preterm birth, low birth weight and late miscarriage. DESIGN: Prospective study in a single centre. SETTING: Guy's and St Thomas' Hospital Trust, London, UK between August 1998 and July 2001. SUBJECTS AND METHODS: Pregnant women were recruited on attending an ultrasound scan at approximately 12 weeks of pregnancy. Subjects completed a questionnaire and underwent periodontal examination. MAIN OUTCOME MEASURES: Plaque and bleeding scores, pocket probing depth and loss of attachment. Pregnancy outcome data was collected retrospectively, including gestational age and birth weight at delivery. RESULTS: Data were collected for 3,738 subjects. Regression analysis indicated that there were no significant relationships between the severity of periodontal disease and either preterm birth (PTB) or low birth weight (LBW). In contrast, there did appear to be a correlation between poorer periodontal health and those that experienced a late miscarriage. CONCLUSIONS: There was no association between either preterm birth or low birth weight and periodontal disease in this population. There is evidence of a correlation between markers of poorer periodontal health and late miscarriage. PMID- 15359325 TI - The quality of dental casts used in crown and bridgework. AB - OBJECTIVE: To assess the quality of dental casts used in crown and bridge construction. DESIGN: Observational cross-sectional study of dental casts. SETTING: Commercial dental laboratories and a university dental hospital laboratory in the UK. MATERIALS AND METHODS: A sample (n = 150) of working and opposing casts used for crown and bridgework prescribed by general dental practices and a dental hospital were sampled from two commercial dental laboratories and an 'on-site' university dental hospital laboratory respectively. A simple '3 point' assessment scale of quality (good, fair and poor) was used to categorise the casts depending on the clarity of reproduction of soft and hard tissues. RESULTS: The quality of opposing casts used for articulation purposes was significantly better (P<0.001) than that of the working casts. In addition it was found that for working casts the quality in the preparation area(s) was significantly better (p<0.001) than that in areas remote from preparation(s) in the same arch. In general, the quality of casts in the incisal or occlusal surfaces was better than the buccal and lingual surfaces. CONCLUSIONS: This study has demonstrated that variation exists in the quality of casts used in crown and bridgework, specifically those used in the construction of indirect restorations and also those used for articulation purposes. This study highlights the need for clinicians to exercise continued vigilance with crown and bridge impressions, and casts, particularly in areas away from the prepared teeth. PMID- 15359326 TI - Undergraduate orthodontic teaching in the new millennium--the Newcastle model. AB - This paper identifies the main factors which have influenced the nature of the undergraduate orthodontic curriculum presently taught at Newcastle University. It also outlines the philosophy that underpins the teaching, states the professional aims of the tutors and spells out the clinical objectives set for the students. PMID- 15359327 TI - Dental treatment in Anglo-Saxon England. AB - Anglo-Saxon medical literature suggests that care of the teeth was largely limited to non-invasive treatment. Cures, mainly for toothache, were based on herbal remedies, charms and amulets. Herbal potions also treated oral problems as diverse as caries and facial deformities. However, surgical intervention for cleft lip is mentioned in the later documents. Skeletal evidence confirms that cleft lip and palate was definitely known to the Anglo-Saxons. A possible dentist's grave has been identified. PMID- 15359341 TI - Protein-modified nanocrystalline diamond thin films for biosensor applications. AB - Diamond exhibits several special properties, for example good biocompatibility and a large electrochemical potential window, that make it particularly suitable for biofunctionalization and biosensing. Here we show that proteins can be attached covalently to nanocrystalline diamond thin films. Moreover, we show that, although the biomolecules are immobilized at the surface, they are still fully functional and active. Hydrogen-terminated nanocrystalline diamond films were modified by using a photochemical process to generate a surface layer of amino groups, to which proteins were covalently attached. We used green fluorescent protein to reveal the successful coupling directly. After functionalization of nanocrystalline diamond electrodes with the enzyme catalase, a direct electron transfer between the enzyme's redox centre and the diamond electrode was detected. Moreover, the modified electrode was found to be sensitive to hydrogen peroxide. Because of its dual role as a substrate for biofunctionalization and as an electrode, nanocrystalline diamond is a very promising candidate for future biosensor applications. PMID- 15359342 TI - Actin-based metallic nanowires as bio-nanotransporters. AB - The synthesis of conductive nanowires or patterned conductive nanoelements is a challenging goal for the future fabrication of nanoscale circuitry. Similarly, the realization of nanoscale mechanics might introduce a new facet to the area of nanobiotechnology. Here we report on the design of conductive and patterned actin based gold nanowires, and on the ATP-driven motility of the nano-objects. The polymerization of G-actin labelled with Au nanoparticles, followed by the catalytic enlargement of the nanoparticles, yields gold wires (1-4 microm long and 80-200 nm high) exhibiting high electrical conductivity. The polymerization of the Au nanoparticle/G-actin monomer followed by the polymerization of free G actin, or alternatively the polymerization of the Au-nanoparticle-labelled G actin on polymerized F-actin followed by the catalytic enlargement of the particles, yields patterned actin-Au wire-actin or Au wire-actin-Au wire nanostructures, respectively. We demonstrate the ATP-fuelled motility of the actin-Au wire-actin filaments on a myosin interface. These actin-based metallic wires and their nanotransporting funcionality introduce new concepts for developing biological/inorganic hybrid devices. PMID- 15359343 TI - Predicting the structure of screw dislocations in nanoporous materials. AB - Extended microscale crystal defects, including dislocations and stacking faults, can radically alter the properties of technologically important materials. Determining the atomic structure and the influence of defects on properties remains a major experimental and computational challenge. Using a newly developed simulation technique, the structure of the 1/2a <100> screw dislocation in nanoporous zeolite A has been modelled. The predicted channel structure has a spiral form that resembles a nanoscale corkscrew. Our findings suggest that the dislocation will enhance the transport of molecules from the surface to the interior of the crystal while retarding transport parallel to the surface. Crucially, the dislocation creates an activated, locally chiral environment that may have enantioselective applications. These predictions highlight the influence that microscale defects have on the properties of structurally complex materials, in addition to their pivotal role in crystal growth. PMID- 15359344 TI - Understanding the phase-change mechanism of rewritable optical media. AB - Present-day multimedia strongly rely on rewritable phase-change optical memories. We demonstrate that, different from the current consensus, Ge(2)Sb(2)Te(5), the material of choice in DVD-RAM, does not possess the rocksalt structure but more likely consists of well-defined rigid building blocks that are randomly oriented in space consistent with cubic symmetry. Laser-induced amorphization results in drastic shortening of covalent bonds and a decrease in the mean-square relative displacement, demonstrating a substantial increase in the degree of short-range ordering, in sharp contrast to the amorphization of typical covalently bonded solids. This novel order-disorder transition is due to an umbrella-flip of Ge atoms from an octahedral position into a tetrahedral position without rupture of strong covalent bonds. It is this unique two-state nature of the transformation that ensures fast DVD performance and repeatable switching over ten million cycles. PMID- 15359345 TI - Ultralong single-wall carbon nanotubes. AB - Since the discovery of carbon nanotubes in 1991 by Iijima, there has been great interest in creating long, continuous nanotubes for applications where their properties coupled with extended lengths will enable new technology developments. For example, ultralong nanotubes can be spun into fibres that are more than an order of magnitude stronger than any current structural material, allowing revolutionary advances in lightweight, high-strength applications. Long metallic nanotubes will enable new types of micro-electromechanical systems such as micro electric motors, and can also act as a nanoconducting cable for wiring micro electronic devices. Here we report the synthesis of 4-cm-long individual single wall carbon nanotubes (SWNTs) at a high growth rate of 11 microm s(-1) by catalytic chemical vapour deposition. Our results suggest the possibility of growing SWNTs continuously without any apparent length limitation. PMID- 15359346 TI - Frozen section diagnosis for axillary sentinel lymph nodes: the first six years. AB - Intraoperative frozen section of axillary lymph nodes for the detection of metastatic breast cancer has been controversial because of the labor-intensive techniques necessary to obtain a highly sensitive test, and because of the uncertain significance of frequently detected submicrometastatic carcinoma. In total, 874 consecutive axillary sentinel lymph node cases examined by intraoperative frozen section over a 6-year period were reviewed retrospectively. Frozen section had a sensitivity of 60% and was 100% specific, but when cases with submicrometastatic tumor cells were considered negative, the sensitivity rose to 83%. Rare cases were called 'atypical' on frozen section; almost all of these cases were negative for carcinoma on permanent sections. PMID- 15359347 TI - The population's aging: a challenge, an obligation, and an opportunity in Latin America. PMID- 15359348 TI - Dietary fiber intake and dental health status in urban-marginal, and rural communities in central Mexico. AB - INTRODUCTION: Oral health status in older people is frequently poor which can contribute to inadequate dietary patterns and nutrition status. OBJECTIVE: To investigate whether an association between the number of teeth present and dietary fiber intake exists in elderly people living independently, across different geographic and socioeconomic locations. METHODS: A cross-sectional household survey was undertaken in three Mexican communities (urban, marginal urban, and rural), incorporating geographic and socio-demographic information and a 24-hour diet recall. Dental status (teeth present, coronal and root caries, and periodontal status) was determined by clinical examination. Data were analyzed using ANOVA, Pearson's chi2 and Tukey's range tests. RESULTS: 407 persons 60 years old and over participated in the study. Subjects in the rural community had better dental/periodontal status and more teeth present than urban and marginal urban participants. Intake of fiber was 8.4 g/day for the urban, 7.6 g/day for the marginal-urban, and 13.5 g/day for the rural community. While gender had no detectable effect, the location of residence and the number of teeth present were associated with mean fiber intake; having more than 21 teeth and/or living in a rural location were associated with increased mean fiber intake. PMID- 15359349 TI - Gait velocity in senior people. An easy test for detecting mobility impairment in community elderly. AB - BACKGROUND: Functional assessment is an important part of the evaluation of elderly patients. Mobility problems detected by functional tests predict the development of more severe disability and injurious events such as falls and hip fractures. Several tests to evaluate mobility have been described, but most of them are difficult to perform by a primary care physicians or take much time in the ambulatory setting. PURPOSE: To evaluate different mobility test to detect mobility impairment in community senior people. Select an easier test to perform on the ambulatory ward by a GP with the hypothesis that gait velocity could be an easier test to detect early mobility impairment. METHODS: A cohort of 100 elderly subjects of 75 year and older were selected from our database and contacted by phone. The subjects were appointed and assessed by three geriatricians from January to May 2000. The measures including MMSE, Yesavage Test, ADL (Barhtel) and IADL (Lawton), the Get Up and Go test, the POMA, one leg balance test and the Gait Velocity (GV). A gait velocity of 0.8 m/s or lower was defined as a pathological gait velocity (PGV). RESULTS: 95 subjects, mean age 79.63 (+/- 4) ranged form 75 to 95. Women in 71.3%. The ADLs were normal on 85% of the patients and the MMSE was normal on 78%. There was a significant association between pathological gait velocity (<0.8m/sec) and impairment on Get up and Go (OR 2.20; 95% CI 1.44-3.34), incapacity to perform the one leg balance test (OR 2.20; 95%CI: 1.43 - 4.71) and abnormal POMA test (OR 4.60; 95 %CI 1.5-14.7). Only 15% of the subjects with normal gait velocity reported recurrent falls in the previous 6 months while 35% of subjects with pathological gait velocity did. (OR 0.32 CI95% 010-099 p < 0.044). CONCLUSION: The pathological gait velocity (<0.8m/sec) correlates with a pathologic performance of Get Up and Go test and POMA and with the incapacity to perform the One Leg Balance test. Also correlate with previous repeated falls in the last 6 (p <0.04). The gait velocity could be a test easy to perform, no time consuming, and an operative tool to apply in the ambulatory care to detect elderly patients with mobility impairment. PMID- 15359350 TI - Determination of body composition using air displacement plethysmography, anthropometry and bio-electrical impedance in rural elderly Mexican men and women. AB - BACKGROUND AND AIMS: Air displacement plethysmography with the BOD-POD is a new densitometry technique, and has been found to be an accurate method to measure body composition. The aim of this study was to assess body composition in a group of free living healthy subjects 60 years of age or older from a rural area of Northwest Mexico, and to evaluate body composition by anthropometry and bioelectrical impedance (BIA) using equations reported for other elderly populations. METHODS: Body composition was evaluated in 21 women and 26 men with the BOD-POD, by anthropometry, and BIA and compared to different equations using the Bland- Altman procedure. RESULTS: Body fat in elderly women and men was 42.7 and 30.2 % respectively. In women, Segal's equation using BIA 1, based on standing height, showed no significant differences with the BOD-POD. There was satisfactory agreement between Segal's equation and BOD-POD. In men similar results was found with Deurenberg's equation using B1A 2 based on recumbent length. CONCLUSIONS: Segal's and Deurenberg's equations were accurate and precise for the estimation of body fat in the elderly of this study. The use of these equations could improve the estimation of body composition for similar elderly subjects. Further studies are recommended with a larger size samples to include elderly subjects from other Latin American countries. PMID- 15359351 TI - Anthropometric measurements of a sixty-year and older Mexican urban group. AB - In the Third World Countries, little attention has been paid to health and nutrition aspects of the elderly population. In Mexico, there are no data that provides anthropometric parameters of this group. The purpose of this study was to obtain anthropometric measurements of 60-year-old-and older Mexican men and women in Mexico City. A cross sectional study was carried out. The sample was selected from men and women registered as retired or pensioned by the Mexican Social Security Institute (IMSS) and from those requesting identification cards from the Elderly National Institute (INSEN). Standardized protocols were used to register anthropometric measurements. The group examined included 1091 people, 484 males and 607 females. The mean age of the population was 66.1 (s.d. 6.1). The values in the male group were higher than in the female group in height, weight and waist circumference; women showed higher values in body mass index (BMI), arm circumference, triceps skinfold and hip circumference (p < 0.01). The data gathered up were divided in five age groups; each one in a five-year interval. Percentiles of the anthropometric measurements according to the age group and gender are presented. Regression analysis indicated that the measurements of weight, body mass index, arm circumference and arm muscle area, showed lower values in the older groups. An important segment of the population studied had a BMI higher to the normal values. Additional studies covering other communities in Mexico with a different socioeconomic and ethnic composition, would be necessary to obtain a better characterization of the Mexican elderly. PMID- 15359352 TI - Validity of height and weight self-report in Mexican adults: results from the national health and aging study. AB - BACKGROUND: An adequate nutritional status is essential for maintaining the independence in the elderly. The height and weight self-report is considered a useful alternative for the estimation of body mass index (BMI). The validity of the self-report is an issue that has not been dealt with in developing countries. AIM: To assess the validity of the height and weight self-report in adults Mexican citizens. DESIGN: Transversal study. STUDY POPULATION: 1707 persons (836 males, 871 females) were asked for their height and weight and were measured. RESULTS: Mean (+/- standard deviation) age was 59.09 +/- 9.86 SD; mean years of education was 5.51 +/-4.67 years. We found a high correlation between self reported and measured weight (R2= 0.837); the difference between both values rose along with the age (from 0.4 kg to 1.74 kg). A systematic difference between self reported and measured height was found. Self-reported height was over-estimated, and the bias increased along with the age of the subjects (from 1.57 cm to 2.57 cm); further, over-estimation was larger in female individuals (+2.22 cm in female vs. +1.21 cm in male subjects). We calculated a linear model that predicts real height from self-reported height with moderate, although statistically significant results (R2= 0.39 y 0.50, for female and male, respectively, p < 0.0001). Knee height was also used to estimate "adulthood height" and thus, BMI. This method showed age-related dissimilarities, and the linear regression model yielded an unacceptably low correlation (R2= <0.10). The best method to estimate real BMI was to consider self-reported parameters. DISCUSSION: Height and weight estimation using self-reported parameters is an acceptable method. Its precision is not so high in subjects > 75 years. Height over-estimation is an expected finding congruent with age-related corporal changes. The estimation of height using knee height is not a useful method. CONCLUSION: Height and weight self report is a valid method that may be used to accurately estimate height and weight in Mexican people. PMID- 15359353 TI - Incidence of proximal femur fractures in Marilia, Brazil. AB - BACKGROUND: SETTING: Three general hospitals in the town of Marilia that have an orthopaedic and traumatologic unit. Marilia is a Municipality with 161.000 inhabitants in the middle-east of Sao Paulo State, Brazil. PATIENTS/PARTICIPANTS: All inpatients, living in Marilia-SP, aged 20 years or more, with a diagnosis of proximal femur fracture (WHO, International Classification of Diseases, 9th.ed., code 820), in the period of January 01, 1994 and December 31, 1995. MAIN OUTCOME MEASURES: The incidence rates of the proximal femur fractures in Marilia-SP. Secondary Measurements: mean-age of the occurrence (male and female), in-hospital mortality, hospitalar costs to S.U.S. (Government Health System), the average length of hospital stay, seasonality, mean-interval between admission and surgical procedure, type of fracture: transcervical and pertrochanteric, content validity of S.I.H.-S.U.S data base report on proximal femur fractures, when compared with hospital registrations. OBJECTIVE: To determine the incidence (crude, age-specific and age-adjusted) of fractures of the proximal femur in Marilia-SP, Brazil, in 1994 and 1995. DESIGN: Retrospective cohort study. RESULTS: The crude incidence rate was 4.96/10,000 inhabitants/year in 1994 and 5.51/10,000 inhabitants/year in 1995; the age-specific incidence rate increased from 0.25/10,000 inhabitants 20-49 years/year to 100.27/10,000 inhabitants 70 years or more/year in 1995 among women; the age-adjusted incidence rate was 29.48/10,000 inhabitants 60 years or more/year in 1994, and 35.83/10,000 inhabitants 60 years or more/year in 1995. CONCLUSION: The crude incidence rate of the proximal femur fractures in Marilia-SP, Brazil was 4.96 / 10,000 inhabitants in 1994 and 5.51/10,000 inhabitants in 1995. It was significantly greater among women (7.2/10,000 inhabitants in 1994 and 8.6/10,000 inhabitants in 1995) and among the elderly, 70 year-old or more (female: 90.21/10,000 inhabitants in 1994 and 100.27/10,000 inhabitants in 1995; male: 25.46/10,000 inhabitants in 1994 and 45.66/10,000 inhabitants in 1995). PMID- 15359354 TI - The Bambui health and aging study: is calibration of dietary intake necessary among older adults? AB - Accurate measurements of food intake in surveys are difficult to be obtained especially in the older adults. Calibration, using two instruments, has been adopted in order to improve the information. This study was carried out in a random sample of older adults (> or =60) living in a Brazilian town. Food intake was obtained by the semi-quantitative food frequency questionnaire associated with photos (SFFQ-P) and the 24 hours recall (R24) with or without food models. Statistical analysis included tests to compare averages, Pearson's correlation coefficient and multiple linear regression. For all nutrients, the average intake obtained through SFFQ-P was significantly higher compared to the obtained by the R24 (p < or =0.05). SFFQ-P correlated better with R24 with food models, which was chosen as the reference method. In the multiple linear regression, vitamin C, fatty acid, protein and zinc intakes were influenced by age, especially when such nutrients were more concentrated in food not frequently reported. For protein and zinc, the significantly interaction between age with the 24 hours recall may have happened because of a differential selection of food sources as a function of age. This fact is probable related to chewing difficulties, reduction in the income and, a diet simplification associated with less complicated preparations, such as meat. This study reinforces the need for calibration of SFFQ in nutritional surveys among older adults, especially in etiological studies where the exposure assessment has to be accurate. In this case, R24 with food models should be used as a reference method to best estimate the true intake. PMID- 15359355 TI - Lean and fat mass as determinants of muscle strength and insulin sensitivity in Chilean elderly subjects. AB - PURPOSE: To report the association of lean body mass with nutritional, social and economic factors and its functional consequences in free living healthy elderly subjects. MATERIAL AND METHODS: Healthy elderly subjects of low socioeconomic level were studied. Monthly income, marital status, anthropometric measures and fall risk were assessed. Mini Nutritional Assessment score was calculated. Body composition and bone mineral density were measured by double beam X ray absorptiomentry. Fasting serum lipids, fasting and postprandial insulin and glucose levels were measured. Hand grip, quadriceps and biceps strengths and maximal inspiratory and expiratory pressures were measured. RESULTS: One hundred and nine subjects (56 women), aged 75 +/- 4 years old were studied. Lean body mass was 34.1 +/- 4 and 49.2 +/- 5.4 kg in women and men respectively (p < 0.001), fat mass was 22.8 +/- 7.1 and 20.7 +/- 6.4 kg in women and men respectively (p= NS). Lean body mass correlated with hand grip, quadriceps and biceps muscle strengths in men and with quadriceps and biceps strength in women. Men that exercised regularly had higher quadriceps strength and maximal expiratory pressure. Total body fat correlated positively with fasting and postprandial serum insulin levels. CONCLUSIONS: In this sample, lean body mass is directly related to muscle strength mostly in men. On the other hand, total fat mass is related to serum insulin levels. PMID- 15359356 TI - Biological effects of drinking-water mineral composition on calcium balance and bone remodeling markers. AB - OBJECTIVE: To compare the effects of 2 drinking waters containing similar calcium (Ca) concentration in order to analyze the role of ions other than Ca on bone metabolism. These mineral drinking-waters differed by their mineral composition primarily concerning the concentration of bicarbonate (HCO3-), high in the HB, and sulfate, high in HS water. DESIGN: Of 60 included women, 39 completed the study. Patients were randomly assigned to an intake of 1 liter per day of mineral water HB or HS for 28 d, followed by cross-over to the alternative drinking-water for a further 28 d. At baseline and after each period of one month, Ca metabolism parameters, acid-base status, and bone remodeling markers were measured. RESULTS: Changes in Ca metabolism were significant in the HB group where the ionized Ca increased and the PTH decreased. Serum pH showed a similar increase whatever the used drinking water compared to baseline. In the HB group, significant increase in urine pH, and significant decrease in AT-HCO3- and NH4+ were observed. Bone resorption markers, urinary CTx/Cr, Pyr/Cr, and D-Pyr/Cr, significantly decreased in the HB group compared to baseline, and were not significantly modified in the HS group. CONCLUSIONS: These results showed a beneficial effect of the bicarbonaterich HB water on bone metabolism. This may account for a better bioavailability of the Ca, a greater alkalinization, and a larger decrease in PTH level secondary to a higher ionized Ca level. The higher content of silica in HB water may have also participated to the positive action on bone balance that was observed. In this short term study, these data underlined the potential role of the mineral drinking water composition on bone metabolism. PMID- 15359357 TI - Validity of measuring knee-height as an estimate of height in diseased French elderly persons. AB - Body mass index is a tool to screen undernourished persons. Cut-off values of 22 or 24 kg/m2 are used for elderly persons. However, standing height cannot be measured in every elderly person. Knee-height has been proposed to estimate standing height, with equations derived from North-American people. The present study analyses the validity of these equations. Standing height and knee-height were measured in 126 elderly persons hospitalized in 6 French geriatrics wards. Consequences of using one of the either height was evaluated on body mass index, body surface area, and total body water as estimated from bioelectrical impedance measurements. Standing height and height derived from knee-height differed significantly by 0.9 cm (P=0.05). This has very limited consequences on the calculation of BMI (0.5 kg/m2), body surface area (0.04 m2) and on total body water (0.2 litres). A regression model was derived from covariates and found the same features as that in the model of Chumlea and Guo. In conclusion, knee-height can be used with sufficient accuracy as a surrogate for standing height in diseased French elderly persons. PMID- 15359358 TI - Skipping of meals has a significant impact on dietary intake and nutritional status of old (65+ y) nursing home residents. AB - BACKGROUND: In spite of a high prevalence of undernutrition among old nursing home residents, studies have generally reported a mean intake of energy sufficient to cover the mean estimated energy requirement. This discrepancy could be due to skipping of meals and hence periods of insufficient energy intake too intermittent to be identified, when mean results are presented. OBJECTIVE: To examine the significance of skipping of (part of) meals on dietary intake and nutritional status of old (65+y) nursing home residents. DESIGN: Participants were 132 nursing home residents (84 (82-85) y). They were characterised according to activity of daily living-functions (ADL-functions), cognitive performance, intake of energy and protein (4-days dietary record), skipping of meals, energy and protein balance, and nutritional status (body mass index (BMI)). RESULTS: The participants had an energy intake, which was significantly higher than the estimated energy requirement (p <0.001). However, 41 (31%) had nothing to eat or only had desert at one or more meals during the 4-day registration period. Participants who were skipping meals had a lower BMI, energy and protein intake (all p <0.001) and a higher prevalence of negative protein balance (p <0.01), than the other residents. CONCLUSION: More focus should be given to individualized nutritional assessment in order to discover the cause to this problem and target individuals who could be helped by nutritional intervention. PMID- 15359359 TI - Macronutrient intakes of elderly in the Lipid Research Clinics Program Prevalence Study. AB - As part of the Lipid Research Clinics (LRC) Program Prevalence Study of coronary heart disease risk factors, nutrient intakes of two US populations over 59 years of age were determined by 24-hour recalls in the 1970s. Characteristics of the populations were (1) California: suburban, upper-middle class, 95% high school graduates, 10% blue collar occupations; (2) Oklahoma: rural lower-middle class, 75% high school graduates, and 40% blue-collar occupations. Macronutrients consumed by both populations were similar, except for alcohol. For both men and women, energy intake was approximately 25 kcal/kg/day (body weight) sources of energy were approximately 40% from carbohydrate, 16% from protein, 37% from fat, and 4% from alcohol. The Oklahoma population, however, consumed significantly less alcohol than did Californians. Percentages of calories from fatty acids were approximately 14% from saturated and 6% from polyunsaturated, which yielded a polyunsaturated: saturated ratio of 0.48. The intake of cholesterol for women was 190 mg/1000 kcal and for men, 210 mg/1000 kcal. Between the ages of 60 and 69, the Oklahoma men consumed more energy than did the California men. Both sexes demonstrated lower energy intakes with advancing age and with increasing body mass index. The higher energy intake of the Oklahoma cohort aged 60-69 was attributed to the greater physical demands of their occupations, but this difference disappeared after age 70. The California and Oklahoma women had similar caloric intakes beyond age 60. In summary, the LRC findings suggest that geographically diverse American populations consumed in the late 1970s remarkably similar intakes of macronutients and cholesterol, with the only major exception being energy from alcoholic beverages. PMID- 15359360 TI - Bone density, parathyroid hormone, calcium and vitamin D nutritional status of institutionalized elderly subjects. AB - INTRODUCTION: Osteoporosis is characterized by a decrease in bone mass and a change in bone microarchitecture. This causes skeletal fragility and may result in fractures. Fractures of the femur neck (FM) are frequent and have major consequences in terms of morbidity and mortality in elderly subjects. We carried out a cross-sectional study to evaluate bone and nutritional status in an elderly population from a same geriatrics department. AIMS OF THE STUDY: We aimed to screen for nutritional deficiencies and to measure bone mineral density (BMD) in the femur neck by dual-energy X-ray absorptiometry (DEXA). RESULTS: The study included 64 white Caucasian subjects, 44 of whom were women. The mean age of the subjects was 80.6 +/- 7.1 years. The mean time since the menopause was 31.45 +/- 7.8 years and 31 subjects presented previous fractures, 12 of which concerned the FM. Mean body mass index (BMI) was 25.8 +/- 4.4 and mean calcium intake was 670 +/- 258.3 mg/d. Mean PTH level was 48.5 +/- 30.34. Thirty-five subjects presented vitamin D deficiencies (mean concentration 8.56 +/- 5.2 microg/L), and 11 of these patients had associated secondary hyperparathyroidism. Fifty-four subjects had osteoporosis (T score <-2.5 standard deviation). Mean bone mineral density (BMD) was 0.596 +/- 0.157 g/cm2 for the femur neck (T score = -3.15 +/- 1.39 standard deviation) and 0.501 +/- 0.169 g/cm2 for the trochanter (T score = - 2.55 +/- 1.68 standard deviation). CONCLUSIONS: Vitamin D deficiency and low calcium intake were observed in a large number of elderly subjects. The patients with the lowest BMD values had secondary hyperparathyroidism. PMID- 15359361 TI - Reduced concentrations of several vitamins in normal weight patients with late onset dementia of the Alzheimer type without vascular disease. AB - BACKGROUND: There is an uncertainty to what an extent initiation of late onset dementia of the Alzheimer type (DAT) is related to nutritional factors. OBJECTIVE: To find any differences in nutrient concentrations between women (75 85 y), well-nourished with moderate DAT, and a control group, all without vascular disease. DESIGN: A case control study assessing clinical, anthropometrical, biochemical and micronutrient characteristics of 20 DAT patients and 18 free-living healthy women. RESULTS: Significant differences (* = p < 0.05) were found for the following nutrients, given in sequence (Mean (SD)) for controls and DAT patients, respectively: Thiamine (nmol/L): 11.7 (6.9), 7.1(3.7)*; Blood thiamine diphosphate (nmol/L): 86.0 (12.5), 65.8 (27.5)*; Pyridoxal-5-phosphate 90.2 (14), 24.8 (3.3)*; Cobalamin (nmol/L) 435(263), 350 (264)*; Homocysteine (mmol/L) 14.7 (1.3), 18.5 (1.6)*; Ascorbic acid (mmol/L) 77.7 (28), 46.2 (25)*; alpha-tocopherol (mmol/L) 38.2 (9.2), 27.1 (11.5)*; Serum and blood thiamine mono-phosphate and ascorbic acid in cerebrospinal fluid were significantly different as well. Age, BMI, MMSE, MADRS, 'Vascular Score ' and a set of other biochemical parameters were similar between the groups. Using logistic regression analysis, models for predicting the presence of DAT all contained pyridoxal-5-phosphate, and CSF-protein, in combination with either one of variables, age, ascorbic acid, retinol, alpha-tocopherol, homocysteine, thiamin-diphosphate, CSF-thiamin. All the models give complete separation between DAT and controls. CONCLUSIONS: The presence of reduced concentrations of several vitamins in the DAT patients compared to the controls might indicate that these nutrients may contribute to the development of DAT. PMID- 15359362 TI - Five-day food intake in elderly female outpatients with Parkinson's disease, rheumatoid arthritis or stroke. AB - AIM: The aim of this study was to describe and analyse the intake of food, energy and selected nutrients in elderly outpatients, i.e. women with Parkinson's disease, rheumatoid arthritis or stroke. SUBJECTS AND METHODS: Sixty-three elderly women aged 64-88 years participated in the study. For assessing dietary intake, a repeated 24-h recall and an estimated food diary for three consecutive days were used. RESULTS: The mean age was 73.4 +/- 6.6 years. Mean reported daily energy intake was 6.4 +/- 1.7 MJ, i.e. lower than reference figures for all groups. However, looking at nutrient density, only intakes of vitamin E and folate were below recommended levels. The most frequently consumed food groups were bread, coffee, milk products, buns and cookies, and spreads. CONCLUSION: The reported energy intake among the elderly female outpatients was low. This might be explained by actual low intake and/or under-reporting. However, the intake of most vitamins and minerals, i.e. nutrient density, was adequate, with the exception of vitamin E and folate intake, which was below recommended levels. Food intake showed large variation and good diet composition, and there was a tendency towards high consumption of food items that are easily prepared and eaten. PMID- 15359364 TI - Emerging issues and trends in attention deficit hyperactivity disorder: an update for the speech-language pathologist. AB - There are many trends in the diagnosis and treatment of attention deficit hyperactivity disorder (ADHD) that have emerged over the past several years. The first of these emerging trends is the growing prevalence of ADHD diagnoses across age groups and cultures. Although the causes of this growing prevalence are unclear, several suggestions, including a broader definition of ADHD, are discussed as potential contributions. A second emerging issue is related to the underlying theory of ADHD. Barkley's biopsychosocial model of ADHD is briefly discussed as a prototype model to help in the understanding of ADHD. Finally, the growing trend in pharmacological treatment of ADHD is discussed. Stimulant medications, as well as a recent nonstimulant medication are discussed and compared with nonpharmacological alternatives. PMID- 15359365 TI - Attention deficit hyperactivity disorder and concomitant communicative disorders. AB - The relationship between attention deficit hyperactivity disorder (ADHD) and communication disorders is complex and often misunderstood. Misunderstandings arise in part because of definitions and diagnostic categorizations that change over the years and because of overlapping symptoms between diagnostic categories. In addition, many of the symptoms that are necessary for the diagnosis of ADHD are prerequisite skills required to perform well on standardized language tests. Consequently, it is difficult to know the exact prevalence of ADHD and concomitant communicative disorders, especially language disorders. Several hypotheses are developed that indicate why the differential diagnosis is so difficult. PMID- 15359366 TI - Attention deficit hyperactivity disorder and preschool children. AB - This article presents a review of the literature on attention deficit hyperactivity disorder (ADHD) and preschool children. A review of the research revealed a predominant focus on pharmacological intervention, with considerably less research on other intervention approaches to ADHD in preschoolers. In particular, there was limited research related specifically to characteristics and best treatment of language-impaired preschool children with ADHD. The available literature did provide important information to further the understanding of the nature of ADHD in preschool-age children and to guide more effective assessment and intervention, both in a general sense and in the practice of speech-language pathology. The more general features of ADHD in preschool children (nature, diagnosis, intervention) are presented first, and then language service delivery implications are discussed. There is limited literature to guide language assessment and intervention for preschool children with ADHD; however, extrapolating from the literature on school-age children would suggest a focus on pragmatic skills, social skills, and emergent literacy. PMID- 15359367 TI - Attention deficit hyperactivity disorder: assessment and treatment of children from culturally different groups. AB - Attention deficit hyperactivity disorder (ADHD) is now thought to affect from 3 to 7% of school-age children. Children with ADHD will often experience serious problems functioning in the school environment. More and more children from culturally diverse groups are being diagnosed as having ADHD. Research shows that culture can directly affect both assessment and treatment of ADHD. This article provides an overview of the effects of cultural differences on the assessment and treatment of ADHD. Children from diverse cultural groups may be overidentified as having ADHD. Cultural difference may also have an impact on access to treatment and compliance with treatment programs. Recommendations for practitioners are included. PMID- 15359368 TI - Perspectives on attention deficit hyperactivity disorder: executive functions, working memory, and language disabilities. AB - The conceptualization of the nature of attention deficit hyperactivity disorder (ADHD) has changed in the last decade. ADHD is now viewed as a neurologically based condition with primary deficits in executive functions and working memory (WM). Students with ADHD have deficits in discourse organization, inferring, and monitoring that are related to their executive function and WM deficits. A large number of students with ADHD also have comorbid reading and language disabilities that exist in addition to the deficits directly associated with the ADHD. Comprehensive evaluation of students with ADHD is essential to address their specific learning needs. PMID- 15359369 TI - Inner control as an operational mechanism in attention deficit hyperactivity disorder. AB - Understanding children with attention deficit hyperactivity disorder (ADHD) through a systems theory perspective creates a view of these children as having developed ineffective systems and strategies for acting upon the world. It is proposed that just as a child must develop a system of inner control to become a proficient and independent reader, this same system is necessary for effectively controlling the problematic behaviors associated with ADHD. Speech-language pathologists (SLPs) can encourage the development of this inner control by providing services that foster meaning-based literacy and that enhance visual perception and processing strategies. As SLPs and educators seek to establish and employ embedded literacy activities, they will more effectively meet the needs of their students identified with ADHD. PMID- 15359370 TI - Speech and language therapy in hyperactivity: a United Kingdom perspective in complex cases. AB - In the United Kingdom, multidisciplinary teams within the National Health Service (NHS) usually manage children and adolescents with disorders of hyperactivity. These multidisciplinary teams do not routinely include speech and language therapists (SLTs), and therefore speech and language therapy services for children and adolescents with disorders of hyperactivity are limited. Consequently, research and service delivery for children and adolescents with disorders of hyperactivity in the United Kingdom is only starting to emerge. Two case studies are used to show how speech and language therapy in the United Kingdom can be involved in the treatment of hyperactivity through various types of service delivery. The cases are used to highlight the importance of detecting speech, language, and communication impairments in children with diagnoses of hyperactivity. The potential role of speech and language therapy in the assessment and management of these clients in the United Kingdom is discussed. PMID- 15359371 TI - Attention deficit hyperactivity disorder: an authentic story in the schools and its implications. AB - As a way of discussing several important facets of attention deficit hyperactivity disorder (ADHD) service delivery in the schools, this article employs a brief narrative case study as a pedagogical technique. Six specific strategies are contextually presented to show one way to effectively collaborate in public education. PMID- 15359372 TI - Owning up to complexity: a sociocultural orientation to attention deficit hyperactivity disorder. AB - To enrich our conception of attention deficit hyperactivity disorder (ADHD), it is necessary to take a wider orientation to this disability category than has been advocated traditionally. Over the past decade, there has been an emerging conception of ADHD from a sociocultural perspective, and this orientation, when linked to the traditional biomedical perspective, provides a more accurate and authentic construct of ADHD. In this article, we advocate that speech-language pathologists approach ADHD with a mindset that is open to the complexities of context-bound human functioning at all levels. Four sources of data demonstrating the richness of the sociocultural orientation are presented and clinical implications are detailed PMID- 15359374 TI - Reboxetine induces similar sleep-EEG changes like SSRI's in patients with depression. AB - BACKGROUND: Reboxetine is a novel selective noradrenaline reuptake inhibitor. The antidepressive properties of the substance are established. METHODS: The influence of reboxetine on the sleep-EEG of eight patients with depression HAMD (mean +/- SD) 19.7 +/- 1.5 (5 women, 3 men; age range 31 to 75 years) was investigated. Sleep EEG was examined twice. The first examination was performed before starting active medication. The second examination was subsequently performed when patients received 8 to 10 mg reboxetine per day. RESULTS: Conventional sleep-EEG analysis showed a significant increase in intermittent wakefulness and sleep stage 2 and a decrease in sleep efficiency and REM time. Under reboxetine no significant changes were observed in sleep-EEG spectral analysis. CONCLUSION: Our results indicate, that reboxetine induces sleep-EEG changes similar to those after selective serotonin reuptake inhibitors (SSRI's) by increasing intermittent wakefulness and decreasing REM time. PMID- 15359375 TI - Comparison of the treatment with paroxetine and reboxetine in panic disorder: a randomized, single-blind study. AB - INTRODUCTION: Serotonergic agents have greater effectiveness than noradrenergic ones in the treatment of Panic Disorder (PD). However preliminary studies suggested that reboxetine might be effective in the treatment of PD. We compared the effectiveness and tolerability of reboxetine and paroxetine in the treatment of PD. METHODS: Sixty-eight patients with PD were assigned to treatment groups in a single-blind, randomized design. Each patient was assessed at day 0 and 90 by the Panic Associated Symptoms Scale (PASS), the Sheehan Disability Scale (SDS) and the Fear Questionnaire (FQ). Side effects were also recorded. RESULTS: Reduction of PASS scores was significantly greater in the paroxetine group than in the reboxetine one. Vice versa we did not find any significant differences for other outcome measures. Sexual dysfunction and weight gain were significantly less frequent in the reboxetine group. CONCLUSIONS: The results showed a greater effect of paroxetine on panic attacks than reboxetine, while no differences for anticipatory anxiety and avoidance were found, suggesting a different role of noradrenaline and serotonin in the treatment of PD. PMID- 15359376 TI - Polysomnography during withdrawal with clomethiazole or placebo in alcohol dependent patients--a double-blind and randomized study. AB - INTRODUCTION: The present study was designed to assess if the routine application of clomethiazole to ameliorate withdrawal symptoms in chronic alcohol dependent patients perpetuates sleep disturbances. METHODS: Twenty inpatients with alcohol dependence according to DSM-IV criteria received clomethiazole or placebo in a double-blind, randomized design upon admission. 11 patients were randomized to the clomethiazole group and 9 patients to the placebo group. During the first 5 days of treatment the patients received either clomethiazole (1st day: 3 x 384 mg, 2nd day: 4 x 384 mg, 3rd day: 3 x 384 mg, 4th day: 2 x 384 mg and 5th day: 1 x 384 mg) or placebo capsules at constant intervals. The patients spent two consecutive nights in the sleep laboratory at each of three assessment times: the first time at the beginning of abstinence (night 1 and 2, T0), the second time 6 days later (i. e. after 5 days of treatment and one day of discontinuation of clomethiazole or placebo: nights 6 and 7, T1) and the third time after 13 days (nights 13 and 14, T2). The first night at each of the three assessment times was an adaptation night. RESULTS: During the first two weeks of abstinence, the analysis of variance demonstrated a significant variation of Rapid Eye Movement (REM) sleep variables in the clomethiazole group. The placebo group showed no such variation. Clomethiazole evidently had a pronounced REM sleep suppressing effect, whereas the discontinuation of clomethiazole led to a REM sleep rebound. Furthermore, analysis of sleep continuity and sleep architecture variables showed that the clomethiazole group had significantly disturbed sleep at T1 in comparison to the placebo group. Simultaneous statistical testing with alcohol intake as covariate reduced the test power so that contrasts between the groups became nonsignificant. CONCLUSIONS: The REM sleep results are in line with earlier findings that REM sleep disinhibition in primary alcohol dependency is partly due to a REM sleep rebound after withdrawal from medication. Differences in the polysomnographic variables of sleep continuitiy and sleep architecture at T0 and T1 found between the clomethiazole and the placebo patients correspond to rebound insomnia following discontinuation of clomethiazole. Our findings indicate that drugs enforcing GABAergic neurotransmission may perpetuate the neuroadaptative effects caused by chronic alcohol consumption. PMID- 15359377 TI - Repetition of serotonin syndrome after reexposure to SSRI--a case report. AB - We describe here the occurrence of a serotonin syndrome (SS) in a 64-year-old depressed female patient with alcoholic hepatic cirrhosis after treatment with SSRIs. Two weeks after the increase of the dosage of sertraline, the patient developed a full-blown SS, which resolved completely after the discontinuation of the drug. The therapy with citalopram led again to development of milder SS, this time immediately after the increase of the dosage. Our case illustrates the variability of the clinical presentation and the temporal evolution of SS in a patient with preexisting medical illness affecting hepatic metabolism. Reexposure of patients with a history of SS to another serotoninergic drug should be avoided; if necessary, it must be carried out with the utmost caution. PMID- 15359378 TI - The central effect of chromium on glucose metabolism. PMID- 15359379 TI - Isolated 3-methylcrotonyl-CoA carboxylase deficiency: evidence for an allele specific dominant negative effect and responsiveness to biotin therapy. AB - Deficiency of 3-methylcrotonyl-CoA carboxylase (MCC) results in elevated excretion of 3-methylcrotonylglycine (3-MCG) and 3-hydroxyisovaleric acid (3 HIVA). MCC is a heteromeric mitochondrial enzyme comprising biotin-containing alpha subunits and smaller beta subunits, encoded by MCCA and MCCB, respectively. Mutations in these genes cause isolated MCC deficiency, an autosomal recessive disorder with a variable phenotype that ranges from severe neonatal to asymptomatic adult forms. No reported patients have responded to biotin therapy. Here, we describe two patients with a biochemical and, in one case, clinical phenotype of MCC deficiency, both of whom were responsive to biotin. The first patient presented at 3 months with seizures and progressive psychomotor retardation. Metabolic investigation at 2 years revealed elevated excretion of 3 MCG and 3-HIVA, suggesting MCC deficiency. High-dose biotin therapy was associated with a dramatic reduction in seizures, normalization of the electroencephalogram, and correction of the organic aciduria, within 4 weeks. MCC activity in fibroblasts was 25% of normal levels. The second patient, a newborn detected by tandem-mass-spectrometry newborn screening, displayed the same biochemical phenotype and remained asymptomatic with biotin up to the age of 18 months. In both patients, sequence analysis of the complete open reading frames of MCCA and MCCB revealed heterozygosity for MCCA-R385S and for the known polymorphic variant MCCA-P464H but revealed no other coding alterations. MCCA R385S is unusual, in that it has a normal amount of MCC alpha protein but confers no MCC activity. We show that MCCA-R385S, but not other MCCA missense alleles, reduces the MCC activity of cotransfected MCCA-wild-type allele. Our results suggest that MCCA-R385S is a dominant negative allele and is biotin responsive in vivo. PMID- 15359380 TI - Genetic variation in radiation-induced expression phenotypes. AB - Studies have demonstrated that natural variation in the expression level of genes at baseline is extensive, and the determinants of this variation can be mapped by a genetic-linkage approach. In this study, we used lymphoblastoid cells to explore the variation in radiation-induced transcriptional changes. We found that, among normal individuals, there is extensive variation in transcriptional response to radiation exposure. By studying monozygotic twins, we demonstrated that there is evidence of a heritable component to this variation. The postradiation variation in the expression level of several genes, including the ferredoxin reductase gene (FDXR) and the cyclin-dependent kinase inhibitor 1A gene (CDKN1A), is significantly greater (P<.001) among twin pairs than within twin pairs. The induction of FDXR by radiation showed a bimodal distribution. Our findings have important implications for understanding the genetic basis of radiation response, which has remained largely unknown due to the lack of family material needed for genetic studies. Our approach, which uses expression phenotypes in cell lines, allows us to expose cells from family members to radiation. Similar study design can be applied to dissect the genetic basis of other complex human traits. PMID- 15359381 TI - Currarino syndrome: proposal of a diagnostic and therapeutic protocol. AB - BACKGROUND/PURPOSE: The Currarino syndrome (CS) is a peculiar form of caudal regression syndrome (CRS) characterized by the association of hemisacrum, anorectal malformation (ARM), and presacral mass. The authors analyzed retrospectively their series, and they propose a multidisciplinary diagnostic and therapuetic protocol that until now has not been introduced. METHODS: A series of 6 patients with CS is presented. Five of them were treated initially in other centers. None of them had an early diagnosis. All presented associated anomalies; in 50%, Hirschsprung's disease (HD) and other dysganglionoses were present. One patient died of a presacral ectopic nephroblastoma. RESULTS: Depending on the expressivity, 3 types of CS can be identified, complete, mild, and minimal. Dysganglionoses and HD can be considered part of CS. A multidisciplinary diagnostic and therapeutic protocol is presented. Main points are sacrum x-Ray, molecular genetic diagnosis, radiologic evaluation of every member of CS families, magnetic resonance (MR) evaluation of patient spine and pelvis, suction rectal biopsies, and search for associated anomalies. CONCLUSIONS: This protocol could give a valid contribution to the treatment of CS, allowing an early diagnosis and proposing a rational timing of multidisciplinary surgical procedures. Early diagnosis and treatment are essential to avoid morbidity and mortality from an undiagnosed presacral mass. PMID- 15359382 TI - Minimally invasive video-assisted thyroidectomy: report of 16 cases in children older than 10 years. AB - METHODS: From October 1998 to December 2002 in the Department of Surgery at the University of Pisa, 16 patients ages 18 years or younger (14 girls, 87.5%, and 2 boys, 12.5%; range, 11 to 18; mean age, 15 years, 8 months) underwent a surgical operation with video-assisted technique for thyroid pathology. These patients belong to a larger group of 270 patients treated with this technique. Surgical therapy with video-assisted technique was chosen; a lobectomy was used in 15 cases (90.0%) and a total thyroidectomy in 1 case (10%). RESULTS: Two patients (12.5%) underwent a second video-assisted operation to complete the thyroidectomy for a false-negative result at extemporal histologic examination during the first operation. The histologic examination found benign pathology in 14 cases (87.5%) and a malignant lesion (papillary type) in 2 cases (12.5%). No postoperative complications were observed. CONCLUSIONS: The results of the mini-invasive video assisted technique for thyroidectomy in this preliminary experience seem to be equal to those of the traditional open surgical technique (200 surgical operations for thyroid pathology for the ages 18 years or younger). Elective indications of the mini-invasive video-assisted technique are the volume of the nodule and hystological type; this technique cannot be used in cases of voluminous goiter, medullary carcinomas and poorly differentiated carcinomas. The advantage that this technique offers, in addition to a better postoperation period, is an improved esthetic result, which is particularly important in young patients. PMID- 15359383 TI - Ultrasound-based decision making in the treatment of acute appendicitis in children. AB - BACKGROUND/PURPOSE: Imaging techniques are used widely to diagnose appendicitis. However, the negative appendectomy rate remains at about 15%. The authors assessed ultrasound-based decision making in the treatment of acute appendicitis in children. METHODS: The authors prospectively studied 165 consecutive children (3 to 15 years old) evaluated for appendicitis. Diagnosis and treatment were based solely on ultrasound scan findings. Criterion for appendicitis was a diameter exceeding 6 mm. Severity was classified into 4 grades based on the appearance of intramural appendiceal structure. Patients with grades I or II received antibiotic therapy. Patients with grades III or IV underwent appendectomy. RESULTS: Ultrasound scan diagnosed appendicitis in 93 children (grade I, 7; grade II, 17; grade III, 41; and grade IV, 28). All but 2 patients with grades I or II underwent antibiotic therapy without complication. All grades III or IV patients underwent appendectomy. There was no negative appendectomy among 76 appendectomies during this period. Ultrasound-based prediction of severity was correct in 67 cases (88%). Ultrasonography identified other pathology in 39. CONCLUSIONS: Ultrasonography in children cannot only visualize all inflamed appendices but also predict severity of disease. Treatment based entirely on ultrasound scan identified patients who required surgery for severe appendicitis and permitted successful conservative treatment for mild appendicitis. PMID- 15359384 TI - Hepatoblastoma--evolution of management and outcome and significance of histology of the resected tumor. A 31-year experience with 40 cases. AB - BACKGROUND/PURPOSE: The aim of this study was to retrospectively evaluate and compare the clinical features, treatment strategy, pathology, and outcome of all patients with hepatoblastoma treated at an African hospital over a 31-year period (1970 to 2001). METHODS: Forty patients with hepatoblastoma were divided into 3 groups according to the treatment given. Group I (1970 to 1983, 14 patients) had no protocol therapy; group II (1984 to 1988, 6 patients) received protocol treatment according to Children's Study Group (CCSG) guidelines; group III (1989 to 2001, 20 patients) received SIOPEL protocol therapy. All available clinical, surgical, radiologic, and pathologic data were reviewed and analyzed. RESULTS: Overall patient survival was as follows: group I, 14%; group II, 50%, and group III, 80%. Deaths in group II were caused by chemotherapy-induced immunosuppression only. Prognostic data for group III showed that all tumor related deaths could be predicted by identifying multifocal disseminated growth patterns (P =.001) or vascular invasion (P =.001) in resected tumors. Of the 40 diagnostic tumor biopsies performed, 2 significant complications (1 death, 1 intraperitoneal tumor seeding) occurred. Histologic criteria evaluating these biopsies were not predictive of overall survival. CONCLUSIONS: The introduction of protocol therapy has resulted in a marked improvement in survival. Immunosuppression-related sepsis in our setting resulted in unacceptable mortality in patients treated according to CCSG guidelines. A diagnostic biopsy in hepatoblastoma is of value but not without complications. Preoperative chemotherapy followed by complete surgical excision according to International Society of Paediatric Oncology guidelines yields excellent results with a current survival rate of 80%. PMID- 15359385 TI - Long-term follow-up status of patients with neuroblastoma after undergoing either aggressive surgery or chemotherapy--a single institutional study. AB - BACKGROUND/PURPOSE: Before adopting a national protocol of aggressive chemotherapy for patients with advanced neuroblastoma in 1985, the treatment strategies in use varied considerably among the institutions in Japan. An aggressive surgical resection was the treatment of choice at the authors' institution before 1985. To better understand treatment-related complications, the authors retrospectively investigated the long-term morbidity of patients who had reached adulthood. METHODS: Sixteen patients with neuroblastoma currently 18 years of age or older were incorporated into the study. All were non-mass screening cases, and 13 of the patients had undergone aggressive surgery, whereas the remaining 3 cases had received aggressive chemotherapy. RESULTS: In the 13 patients who had undergone aggressive surgical treatment, a unilateral atrophy of the kidney was noted in 2 cases of adrenal and retroperitoneal neuroblastomas, respectively. Dry ejaculation was noted in one case of a giant presacral neuroblastoma. A maldevelopment of the unilateral lower extremity was also noted in another case with a presacral tumor, which had been resected by abdominal and sacral approaches. In the 3 patients who had undergone aggressive chemotherapy, severe hearing loss occurred in 2 cases. However, social adaptation was well preserved in all 16 cases. CONCLUSIONS: Treatment-specific complications were found in both the aggressive surgery and chemotherapy groups. These results should, therefore, be taken into account when developing future treatment protocols. PMID- 15359386 TI - Continent urinary reconstruction in rhabdomyosarcoma: a new approach. AB - PURPOSE: The authors present here 5 cases of continent urinary diversion in rhabdomyosarcoma applying a recently described technique for the Mitrofanoff Principle devised by the authors. METHODS: Two previously irradiated rhabdomyosarcoma patients presenting with residual bladder disease and massive sensitive urinary urgency underwent a transverse colonic reservoir with catheterizable stoma. Two other patients presenting with a Bricker conduit underwent conversion into an ileal reservoir. One patient underwent reconstruction after a cystectomy. RESULTS: All patients were continent and able to perform continent intermittent catheterization. CONCLUSIONS: The technique proved to be feasible for this group of patients. The authors believe that because of its simplicity, it should be an option of continent urinary diversion when the Mitrofanoff Principle is considered. PMID- 15359387 TI - Diagnosing suspected acute nonclassic appendicitis with sulesomab, a radiolabeled antigranulocyte antibody imaging agent. AB - PURPOSE: The aim of this study was to investigate the potential role of a technetium 99m-labeled antigranulocyte murine antibody Fab' fragment (sulesomab) as a diagnostic imaging agent in children with suspected acute nonclassic appendicitis. METHODS: Serial planar images at 15 to 30 minutes, 1, 2, and 4 hours and single-photon emission computed tomography (SPECT) images were acquired after sulesomab injection. In 40 children with suspected acute nonclassic appendicitis, imaging results were confirmed surgically in 21 patients, whereas 19 nonsurgical patients resolved presenting signs and symptoms and were considered to not have appendicitis. RESULTS: Sulesomab imaging had 95% sensitivity, 90% specificity, 95% negative predictive value, and 90% positive predictive value for acute appendicitis. In 78% of patients, sulesomab accurately detected or excluded acute appendicitis and would have changed management plans. No patients had adverse events, and no human antimurine antibody response occurred in 18 evaluable patients. CONCLUSIONS: Sulesomab was well tolerated with no side effects and with no apparent immunogencity. Appendicitis was rapidly and accurately detected. In pediatric patients with suspected nonclassic appendicitis, management decisions incorporating sulesomab imaging provided benefit in separating surgical from nonsurgical patients. PMID- 15359388 TI - Reliability of intraoperative frozen sections in the management of Hirschsprung's disease. AB - BACKGROUND: The use of a 1-stage pull-through for Hirschsprung's disease (HD) is dependent on accurate identification of the normally innervated bowel on intraoperative frozen sections (IOFS). The authors wished to determine the incidence and sources of error during this process. METHODS: All HD patients undergoing IOFS over a 15-year period were reviewed. RESULTS: Three hundred four patients underwent a total of 700 IOFS. In 9 cases (3%), there was discrepancy between IOFS and permanent sections. Two of these were false-positive (ganglion cells incorrectly believed to be present at IOFS); both required a second operation as a result of the error. Seven were false-negative (presence of ganglion cells not recognized at IOFS); none required a subsequent operation, but 2 had a significantly more extensive colonic resection than was necessary. Responsible factors included sampling from the transition zone, freezing artifact, and misinterpretation of ganglion cells in very young patients owing to pathologist inexperience. There was significant variability in the error rate among the 11 pathologists. However, the numbers were too small for statistical analysis to determine whether there was a correlation between the rate of errors and the volume of cases done or years of experience. CONCLUSIONS: Error in reading of IOFS is rare but can have significant repercussions in patient care. Multiple factors, including technical issues and pathologist experience, may have a role in contributing to these errors. PMID- 15359389 TI - Results of a mechanical Duhamel pull-through for the treatment of Hirschsprung's disease and intestinal neuronal dysplasia. AB - BACKGROUND/PURPOSE: The authors analyzed the results of a modified entirely mechanical Duhamel pull-through for the treatment of Hirschsprung' disease or type B intestinal neuronal dysplasia. The aim of the follow-up was to evaluate results of a Duhamel procedure entirely performed with the use of staplers. METHODS: Fifty-six patients were followed up to detect complications and outcome. Patients were evaluated clinically, and, when symptoms were present, a suction biopsy with histochemical analysis, an x-ray contrast enema, or an endoscopy was performed. RESULTS: Early complications developed in 4 cases (7%) including a case of leakage (1.8%). No patient experienced recurrence. During a mean follow up of 49 months, further complications occurred in 16 other patients: 4 adhesive obstructions requiring surgery, 3 strictures, 5 chronic bleeding, 4 fecaloma formation, 2 urinary problems, and 13 postoperative enterocolitis. Four patients (7%) had more than 3 passages of liquid stools per day, and 3 of them who had an ileal pull-through had constant soiling. Thirty-seven children (68.5%) had 1 to 2 bowel movements per day and were continent. Seven (13%) had less than 3 bowel movements per week. Three children who underwent total colectomy were incontinent (5.5%). Three more children (5.5%) have not yet reached the age for continence. Forty-nine parents (87%) judged the outcome satisfactory. CONCLUSIONS: Follow-up of mechanical Duhamel pull-through showed an acceptable long-term outcome. An increased risk of incontinence is present in case of ileal pull-through, whereas constipation is more frequent after partial colon resection, regardless of the type of intestinal neuronal malformation. PMID- 15359390 TI - Portal vein thrombosis in children and adolescents: the low prevalence of hereditary thrombophilic disorders. AB - PURPOSE: The aim of this study was to determine the frequency of thrombophilic disorders in children and adolescents with portal vein thrombosis (PVT) as well as assessing the hereditary character of this disorder. METHODS: A 2-year prospective study was carried out in pediatric PVT patients (n = 14), their parents (n = 25), and an age-matched control group free of liver disease (n = 28). The presence of PVT was assessed by means of Doppler ultrasound scan or angiography. None of the PVT patients presented biochemical or histologic signs of liver disease. RESULTS: The frequency in PVT patients of protein C (PC), protein S (PS) and antithrombin (AT) deficiency was 42.9% (P <.05 v controls), 21.4% (P >.05) and 7.1% (P >.05), respectively. None of the controls or parents of PVT patients presented hereditary PC, PS, or AT deficiency. One PVT patient and one control (P =.999) presented prothrombin G20210A mutation. Homozygous methylenetetrahydrofolate reductase C677T genotype was observed in 3 of 14 (21.4%) PVT patients and in 5 of 28 (17.9%; P =.356) controls. None of these patients presented factor V G1691A mutation. CONCLUSIONS: PC deficiency was frequent in pediatric PVT patients and does not seem to be an inherited condition. The hereditary prothrombotic disorders do not seem to play a vital role in thrombosis in children and adolescents with PVT. PMID- 15359391 TI - Absorption of D(+)-xylose, cobalamin, and folic acid after autologic-allotopic ileum mucosa transplantation in beagles. AB - BACKGROUND/PURPOSE: Ileum mucosa transplantation in a demucosed colon coat was developed as a new method for small bowel elongation. In an animal model, the authors investigated the absorptive capacity of the transplanted mucosa for D(+) xylose, cobalamin (vitamin B12), and folic acid. METHODS: Ileum mucosa was transplanted in a vascularized demucosed segment of transverse colon in 18 beagle dogs. The colon coat-ileum mucosa complex then was integrated in the ileal continuity. Absorptive capacity for D(+)-xylose, cobalamin, and folic acid was measured before and 4 weeks after transplantation. The results were compared and analyzed with the Students' t test for matched pairs. All determined blood values with P values less than.05 were considered to show a significant reduction in the absorptive capacity of the transplanted ileum mucosa. RESULTS: Fifteen minutes after application there was no significant difference in the absorption of D(+) Xylose and cobalamin between normal and transplanted ileum mucosa (P >.1). Absorption of folic acid in the transplanted segment was lower but not significant (P <.1). After 30 minutes D(+)-xylose and cobalamin again showed no difference between the absorptive capacity of normal and transplanted ileum mucosa (P >.1), whereas folic acid continued with the tendency toward an impaired absorption (P <.1). However, after 60 minutes, the difference of the absorptive capacity of the transplanted ileum mucosa was significant (P <.05) for folic acid. D(+)-xylose showed a tendency for an impaired uptake (P <.1), whereas absorption of cobalamin did not differ significantly after transplantation (P >.1). CONCLUSIONS: Experimental autologic-allotopic ileum mucosa transplantation is a feasible new method for small bowel elongation in an animal model. Examination of the absorptive capacity of the transplanted ileum mucosa showed a normal uptake for cobalamin, while there was an impaired absorption of D(+) xylose and folic acid. PMID- 15359392 TI - The maturity of intestinal neomucosa: integrin expression and ultrastructural aspects. AB - BACKGROUND/PURPOSE: The maturity of neomucosa growing on a serosal surface for the treatment of short bowel syndrome still is questionable. The aim of this study was to evaluate the intestinal neomucosa to assess its histologic maturity. METHODS: A 6-cm-long isolated ileal segment (IS) was prepared in 8 Wistar albino type rats. The IS was divided from the antimesenteric side, and 2 intestinal tubes were established, which shared a common wall and a common pedicle. After ileal biopsy sampling for the control group (CG), the IS was fashioned into a mucous fistula. Eight weeks later, all the rats were killed, and the ISs were investigated for neomucosal growth. Sections were prepared with periodic acid shift (PAS) and H & E staining for light microscopy. They also were evaluated by transmission electron microscopy. The microscopic morphology of the 2 groups was evaluated. Immunohistochemical staining was performed to show the expression of the tissue beta1, alpha3 and alpha2beta1 integrin subunits of both the neomucosa (NS) and control group (CG) segments. RESULTS: Sections of the NS showed a well arranged columnar epithelial cell layer with goblet cells that were generally located superficially and with a complete basement membrane. Under the electron microscope, the sections from the NS group showed an epithelial cell layer with proper microvilli of the same height, although they were shorter than those of the CG, and tight intercellular junctions between the epithelial cells. Significant differences between the NS and CG groups were found in the measurements of villus width at base, microvillus surface, and microvillus height. The lamina propria consisted of rich collagen fibers and active fibroblasts in the NS group. In the immunohistochemical staining, although beta1 integrine showed a dense distribution (+++) in the lamina propria, particularly localizing at the depth of the tunica mucosa layer, alpha3 integrin was observed to have a less dense immunoreactivity (++) in both groups. The expression of alpha2beta1 integrin showed slight and dispersed (+) staining. CONCLUSIONS: The NS showed histologic maturity and ultimate structural similarity with the native small bowel mucosa, which provides strong indirect evidence for the proper functioning of the neomucosa. PMID- 15359393 TI - Endotracheal intubations in rural pediatric trauma patients. AB - BACKGROUND/PURPOSE: Evidence from urban trauma centers questions the efficacy of pediatric field endotracheal intubations (ETIs). It is recognized that in the rural environment, discovery, transport delays, and a paucity of pediatric expertise contribute to higher pediatric trauma mortality rates compared with urban environments. The purpose of our study was to determine the effectiveness of field ETI in rural pediatric trauma patients. METHODS: ETI attempts (field, referring hospital, trauma center [TC]) in trauma patients less than 19 years old were included. Prehospital and TC charts, including demographics, injury mechanism, indication, location, person performing, number of attempts, Glascow Coma Scale (GCS), complications from ETI, and outcome, were assessed. RESULTS: Between 1991 and 2000, 105 of 2,907 patients met study criteria. Paramedics, trauma flight nurses (field ETIs), emergency physicians, surgeons, and anesthesiologists performed the ETI. One hundred fifty-five ETIs (1 to 6 per patient) were attempted in 105 children. Fifty-seven percent of the ETIs were attempted in the field, 22% in transferring hospital, and 21% at the TC. Successful intubation on first attempt was 67% (field), 69% (referring hospital), and 95% (TC). Subsequent ETI attempts had failure rates of 50% (field) and 0% (referring hospital, TC). Indication for ETI included fear of losing airway control (37%), closed head injury (36.1%), respiratory rate less than 10 or greater than 40 (11.2%), cardiopulmonary arrest (6.5%), respiratory arrest (4.6%), and airway obstruction 4.6%. Only 9.3% of children could not be oxygenated or ventilated by bag valve mask (BVM) before ETI. Twenty-three percent had complications directly related to ETI (eg, aspiration). The relative risk of an airway complication was 2.5x higher with more then one ETI attempt (P <.05). Four percent of the airway complications occurred in TC, 29% (transferring hospital) and 66% (field, P <.05), respectively. Airway complications and multiple ETIs were associated with transport delay, lower GCS, longer hospital stay, and lower discharge GCS (P <.001) but independent of injury severity score, sex, age, and survival (P >.05). CONCLUSIONS: Multiple ETI attempts are associated with significant complications and may offer limited advantage over BVM and possibly may affect outcome. Indications for field intubations may require review especially in rural pediatric trauma. PMID- 15359394 TI - The roles of free oxygen radicals, nitric oxide, and endothelin in caustic injury of rat esophagus. AB - PURPOSE: The authors aimed to find out the roles of free oxygen radicals, nitric oxide (NO), and endothelin (ET) in caustic injury of rat esophagus. METHODS: Forty-five Wistar albino rats were used to form 6 groups. The study groups are summarized as 1, sham (S; n = 7); 2, sham + L-arginine (SA; n = 7); 3, sham + L NAME (SN; n = 7); 4, injury (I; n = 8); 5, injury + L-arginine (IA; n = 8); 6, injury + L-NAME (IN; n = 8). Normal saline in the sham groups and 50% NaOH in the caustic injury groups were administered to the distal esophagus. Free oxygen radicals and NO were detected by chemiluminescence from tissue samples, and they were correlated with histologic examinations. Tissue ET was measured also with immunohistochemistry. RESULTS: The injury was verified histologically. Free oxygen radical levels were found to be increased as well as NO and ET with the caustic injury (P <.05). L-arginine caused a histologic increase in the injury that was close to statistical significance (P =.08). L-NAME showed no significant effect. CONCLUSIONS: Free radicals, NO, and ET increase in the early phase of caustic esophageal injury. Understanding their early interactions during the caustic injury may help in future therapeutic strategies. PMID- 15359395 TI - Robotically assisted esophago-esophagostomy in newborn pigs. AB - PURPOSE: Repair of esophageal atresia and tracheoesophageal fistula (TEF) in the newborn is challenging when done as an open procedure but only a few surgeons have attempted this with minimally invasive surgery (MIS). Surgical robots that hold MIS instruments have wrists and provide tremor filtration and motion scaling, which might be expected to facilitate complex procedures in small spaces such as an esophageal anastomosis in a newborn. METHODS: Seven newborn pigs weighing 2 to 3 kg were used as a model. The authors performed an esophageal resection and end-to-end anastomosis using the Zeus Microwrist Robotic Surgical System. The authors monitored the following data-Zeus robotic set-up time, operating time, esophageal dissection time, and anastomosis time. After 1 month, the animals were anesthetized to perform esophagram and than were killed. RESULTS: In these very small animals the space available for performing an anastomosis is very limited, approximately 2 cm3. Two pigs each died 18 days postoperatively. One animal could not eat and died during esophageal dilation. The second pig died of unrelated enteritis. Neither had evidence of anastomotic leak at autopsy. The esophagram of the 5 pigs that survived for 30 days showed no narrowing or proximal dilation in 2. In 3 there was some stenosis requiring dilation. Histopathology was done on each operated animal. Two of the anastomoses were well healed, and 5 showed only focal small ulceration. CONCLUSIONS: Robotic assistance facilitates an MIS approach to the upper esophagus even in the limited space of the infant chest. PMID- 15359396 TI - Gastrostomy revision: incidence and indications. AB - BACKGROUND/PURPOSE: Although quite reliable, gastrostomy may require revision. However, there are no reports in the literature specifically delineating identifiable risk factors or circumstances that lead to gastrostomy revision in children with gastrostomy. The purpose of this report was to determine the rate of revision and correlate any factors that may lead to revision. METHODS: A retrospective chart review was performed on 1,042 children who underwent gastrostomy at The Children's Hospital, Denver, Colorado, between 1992 and 2002. Charts of children who underwent gastrostomy were reviewed for pertinent clinical factors and compared with those who required gastrostomy revision. RESULTS: Of the 1,042 children, who had gastrostomies, 67 revisions were required in 61 children (6%). Of the many possible factors that could have had an influence on the revision rate, only fundoplication, percutaneous endoscopic gastrostomy (PEG), migration of the gastrostomy site, and time correlated with the need for gastrostomy revision. CONCLUSIONS: Parents should be made aware that there is a 6% chance that their child's gastrostomy may need revision and that the need for revision may increase with PEG, initial construction before 18 months of age, and the advancing age of the gastrostomy. Surgeons should avoid placing the gastrostomy near the costal margin, making a large gastrostomy exit tract through the abdominal wall and inserting a gastrostomy into the nutritionally depleted pulmonary stressed neurologically challenged child without first attempting to improve the child's nutritional status. PMID- 15359397 TI - Preoperative sonographic evaluation is a useful method of detecting contralateral patent processus vaginalis in pediatric patients with unilateral inguinal hernia. AB - BACKGROUND/PURPOSE: To analyze the value of ultrasound scan (US) in the preoperative identification of a contralateral patent processus vaginalis (CPPV), the authors investigated the presence of CPPV by US in children with clinically diagnosed unilateral inguinal hernias. METHODS: Three hundred forty-eight patients (168 boys and 180 girls) with unilateral inguinal hernia underwent US examination using a 7.5-MHz linear transducer. If a CPPV was visible as a hydrocele owing to the inflow of physiologic ascites into a processus vaginalis on straining, then US scanning was performed while the patient was at rest and while inducing straining by standing or crying. A groin with a hydrocele in the inguinal canal on straining was diagnosed as a CPPV and was explored bilaterally through surgery. The US findings were compared with surgical results. RESULTS: In 348 patients, 78 cases (22.4%) were diagnosed by US as patients with a CPPV; these patients underwent bilateral surgery. Seventy-four of 78 cases (94.9%) were confirmed surgically as patients with CPPV. CONCLUSIONS: A CPPV was detected correctly by US in 74 of 348 patients (21.3%) with clinically diagnosed unilateral inguinal hernias. US is a noninvasive and accurate method for evaluating the presence of a CPPV. PMID- 15359398 TI - Effects of ropivacaine infiltration on cortisol and prolactin responses to postoperative pain after inguinal hernioraphy in children. AB - BACKGROUND/PURPOSE: Painful interventions may have a serious adverse psychological impact, particularly in young patients. Inguinal hernia repair is the most common surgical outpatient procedure performed on infants and children. The aim of this study was to compare the effects of pre- and postincisional infiltration of the surgical area with ropivacaine on cortisol (C) and prolactin (PRL) release and postoperative pain in children undergoing inguinal hernia repair. METHODS: Forty-five school-age children, aged 6 to 10 years, undergoing outpatient inguinal hernia repair under general anesthesia were placed randomly into 3 groups. Preincisional wound infiltration of 3 mg/kg ropivacaine was performed in group I patients before surgery. Postincisional wound infiltration was performed in group II patients after hernia repair but before skin closure, and group III patients (controls) did not received any local anesthetic. In the postanesthesia care unit (PACU), objective pain assessments were performed every 5 minutes using a standardized 10-point objective pain scale. RESULTS: Plasma C concentrations increased at the end of the operation in all groups but significantly only in the control group (P <.001). There was no significant difference between the pre- and postincisional groups with regard to pre- and postoperative C alterations (P >.05). Although plasma PRL concentrations increased significantly at the end of the operation in the control group (P <.001), no significant difference was found between pre- and postoperative values in the infiltration groups (P >.05). The pre- and postoperative plasma PRL differences were significant between only groups I and III (P <.001). CONCLUSIONS: The findings of the current study suggest that wound infiltration with ropivacaine decreases the stress response to surgery and the postoperative pain. PMID- 15359399 TI - Four-triangular-skin-flap approach to umbilical diseases and laparoscopic umbilical port. AB - BACKGROUND/PURPOSE: Surgery of umbilical pathology requires restoration of a normal umbilical appearance. Also, the umbilicus is used increasingly as the entry site during laparoscopic surgery. However, conventional approaches leave obvious scars. A simple alternative approach that creates a natural-looking umbilicus is described. METHODS: The umbilicus is opened by creating 4 isosceles triangular skin flaps. Closure is by suture of the flap apex only, creating scaring that resembles a natural umbilicus. RESULTS: Between November 1996 and March 2003, this technique was used in 204 children with umbilical hernia, 2 children with a small omphalocele, 1 child with a patent omphalomesenteric duct, 2 children with a urachal abscess, and 7 children with an umbilical granuloma. Five children underwent initial trocar insertion during laparoscopic surgery via this approach. All procedures were performed uneventfully. Transient erythema of one flap occurred in 64 patients (29.2%). Infection developed in 10 patients (5.0%) but was treated with oral antibiotics. The postoperative umbilical appearance was satisfactory in all but 5 patients. CONCLUSIONS: The 4-triangular skin-flap approach is useful for umbilical diseases and laparoscopic umbilical port access. PMID- 15359400 TI - Median sternotomy as an exit procedure in a child with massive pericardial tumor. AB - Fetuses can survive with complete airway obstruction caused by placental gas exchange until birth when full ventilatory function is required. The authors present a case in which prenatal scans suggested that adequate ventilation would not be achievable because of the presence of an intrathoracic tumor. An EXIT procedure (exutero intrapartum treatment) was therefore performed, which permitted sufficient lung expansion for adequate ventilation. PMID- 15359401 TI - Biliary atresia associated with hypoplastic left heart syndrome: a case report and review of the literature. AB - Biliary atresia is a cholestatic disorder of infancy that is associated with other anatomic anomalies in approximately 20% of cases. These frequently are defects in situs determination and laterality, causing syndromes of heterotaxy and complex heart defects. The authors describe an infant with both biliary atresia and hypoplastic left heart syndrome (HLHS) in the absence of a laterality defect or evidence of any other morphologic defect. The presence of a hypoplastic left ventricle in a patient with biliary atresia has previously been reported only in the context of a heterotaxy syndrome. The coexistence of these 2 disorders raises etiologic considerations and represents a potential challenge for the surgical treatment of both conditions. PMID- 15359402 TI - Chest wall lipoblastoma in a seven-month-old girl: a case report. AB - Lipoblastoma is a rare benign tumor of adipose tissue seen almost always in infanthood and early childhood. It is typically located in the extremities and the trunk and less frequently in the head-neck region, mediastinum, and retroperitoneum. However, cases of pleural, thoracic wall, and pulmonary lipoblastoma have also been reported in the literature. Lipoblastoma is a tumor with good prognosis despite its potential for local invasion and rapid growth. Our patient is a 7-month-old girl who was brought to our clinic for a rapidly growing mass in the right hemithorax. With the aim of both diagnosis and treatment, the mass was removed by thoracic wall resection and the resultant defect covered with an absorbable polyglactine mesh. The excised mass was diagnosed pathologically as a benign lipoblastoma. In the postoperative period, the thorax wall was stable, and after 24 months of follow-up no recurrence was observed. In the literature, 4 types of thoracic wall lipoblastomas have been reported, but thoracic wall resection has not been applied in any benign lipoblastoma case. To the best of our knowledge, the only benign lipoblastoma case in which a thoracic wall resection was carried out, with its clinical and histopathologic specifications alongside that in the literature, is presented here for the first time. PMID- 15359403 TI - Primary neurilemmoma of the thyroid gland in a 12-year-old girl. AB - A case of a neurilemmoma of the thyroid gland in a 12-year-old girl is reported. The tumor was noted as a asymptomatic left lower neck swelling 9 months before admission. Ultrasound scan showed the solid lesion of predominantly hypoechoic structure measuring 17.2 x 12 x 26 mm within the left lobe. Pathologic examination of the excised lobe was consistent with neurilemmoma with both Antoni A and Antoni B histologic patterns. To date only 14 cases of this nonepithelial tumor of the thyroid gland have been described in the literature, and all occurred in adult patients. This is the first report of thyroid neurilemmoma in a child. PMID- 15359404 TI - Spontaneous resolution of prenatal megalourethra. AB - Urethral obstruction in the fetus is rare. Whereas proximal obstruction most often is caused by posterior urethral valves, causes of distal obstruction are less well recognized and can include urethral atresia, urethral webs, and anterior urethral valves. These latter abnormalities can lead to urinary retention, incontinence, enuresis, spontaneous bladder rupture, and megacystis. The authors present 3 fetuses (gestational age range, 18 to 20 weeks) in whom distal urethral obstruction was suspected by prenatal ultrasonography in the absence of a demonstrable lesion. All 3 experienced spontaneous resolution of the presumed obstruction. On follow-up, all are alive and well with no adverse genitourinary tract sequelae. No postpartum intervention was required. PMID- 15359405 TI - Infantile fibromatosis of the sternocleidomastoid muscle mimicking muscular torticollis. AB - Infantile fibromatosis is a rare entity in children. Although the cervical region is one of the frequent sites of location for this tumor, the diagnosis may be delayed if it occurs in a usual location for a well-known entity, such as muscular torticollis or fibromatosis colli. The authors present an infant with infantile desmoid fibromatosis of the sternocleidomastoid muscle, which has been considered initially as olive of muscular torticollis. PMID- 15359406 TI - Perforation of a congenital umbilical hernia in a patient with Hurler's syndrome. AB - Congenital umbilical hernias in Hurler's syndrome (mucopolysaccharidosis I) are generally treated conservatively, because complications such as incarceration are, rare, and risks involved in surgical correction are high. This case report describes the surgical management of a ruptured umbilical hernia in a 3-year-old child with Hurler's syndrome. Emergency repair of the hernia was performed with primary closure of the fascia; hernia recurrence 6 months later was treated laparoscopically using a PTFE mesh graft with no evidence of re-recurrence. In selected cases of Hurler's syndrome (warning signs of rupture) elective surgical hernia repair may be indicated. PMID- 15359407 TI - Use of the deep inferior epigastric perforator flap in a 15-month-old boy after resection of lymphatic malformation. AB - Lymphatic malformations are benign tumors that arise from congenital defects of the lymphatic system. Despite the fact that all lymphatic malformations develop embryologically, their presence may not be detected until the tumor enlarges from hemorrhage or infection. Typical of these tumors is a slow-growing, asymptomatic phase with the potential for rapid and massive expansion and a tendency to infiltrate into surrounding tissues. These tumors are associated with a high rate of recurrence, especially with incomplete excisions. To minimize the chance of recurrence, a complete resection of a groin microcystic lymphatic malformation was performed on this 15-month-old boy, creating a significant defect in the upper thigh. Immediate coverage was felt to be the best option for reducing any associated morbidity. We used a skin-soft tissue flap supplied by perforators of the deep inferior epigastric artery to repair the wound. Using this technique, the musculofascial structures in the abdominal wall were completely spared, and the skin and fat were transferred on a vascular leash to the desired location. The donor site was closed by approximating the fascial edges without tension. The defect on the thigh was filled with tissue of similar thickness and characteristics, avoiding potential contour irregularities and color mismatch. The deep inferior epigastric perforator (DIEP) flap is a useful option for groin defects and can be applied safely in the pediatric population. PMID- 15359408 TI - Intercostal lung hernia in a 7-year-old boy. AB - The authors report a case of a lateral chest wall lung hernia, which was presumably congenital. Lung herniations are extremely rare, and congenital herniation of the lung through an apparently intact thoracic cage has not yet been reported. The hernia was repaired by the interposition of a polypropylene mesh. The literature of this rare condition is briefly reviewed. PMID- 15359409 TI - Recurrent sigmoid volvulus in a sixteen-year-old boy: case report and review of the literature. AB - The authors describe a 16-year-old boy in whom 3 episodes of sigmoid volvulus (SV) occurred over a period of 7 weeks, each time reduced by endoscopy. The child subsequently underwent a successful sigmoid resection with primary anastamosis. Several months after surgery, he remains free of symptoms and is doing well. A review of the literature illustrates the approach to this problem from ancient times until now. PMID- 15359410 TI - A case of pediatric ovarian dysgerminoma associated with high serum levels and positive immunohistochemical staining of neuron-specific enolase. AB - A 5-year-old girl presented with a painful abdominal mass. Abdominal magnetic resonance imaging (MRI) showed 3 separate masses. Tumor markers including lactate dehydrogenase (LDH), cancer antigen-125 (CA-125), beta-subunit of human chorionic gonadotropin (beta-hCG) and neuron-specific enolase (NSE) were elevated. At operation, the main tumor arose from the left ovary and was associated with torsion, whereas the other lesions were lymph node metastases. A salpingo oophorectomy was performed. Histopathologic examination indicated that the tumor was a dysgerminoma. Immunohistochemicallly, the cells were positive for NSE and placental alkaline phosphatase (PALP) but were negative for CA-125, beta-hCG, S 100, glial fibrillary acidic protein, and vimentin. The elevated serum levels of tumor markers improved dramatically after the operation and chemotherapy. PMID- 15359411 TI - Renal failure as a result of mesenteric cyst. AB - The authors report a rare case of renal failure secondary to abdominal cyst in a newborn girl. The clinical presentation was one of a largely distended abdomen coupled with anuria. The histopathologic and clinical findings suggest mesenteric cyst causing renal failure by mass effect. PMID- 15359412 TI - A case of human botfly infestation in a child who had recently returned from Central America. PMID- 15359413 TI - The role of beta-catenin in regulating angiogenesis in Wilms tumor. PMID- 15359414 TI - Perforated appendicitis in children: is there a best treatment? PMID- 15359415 TI - Spontaneous liver rupture in a child with graft-versus-host disease. AB - Graft-versus-host disease (GVHD) is a common complication in the stem cell transplant (SCT) patient. The skin, gastrointestinal tract, and the liver are the most frequently affected organs. The authors present the first reported case of a spontaneous rupture of the liver in a child with GVHD after SCT. PMID- 15359416 TI - Retroperitoneoscopic drainage of a psoas abscess. AB - Pyogenic psoas abscess in the pediatric age group is a primary condition caused mostly by Staphylococcus aureus. The preferred treatment is percutaneous or surgical drainage under a cover of systemic antibiotics. Laparoscopic drainage scores over open surgery in terms of minimal invasion, shorter hospital stay, better patient comfort, and more complete drainage compared with the percutaneous approach. The authors report a case of a 4-year-old boy with a psoas abscess that was effectively drained laparoscopically through an extraperitoneal approach. PMID- 15359417 TI - Three new cases of disorganizationlike syndrome: one with accessory extrophia vesicalis. AB - The authors report 3 unrelated Turkish cases of disorganizationlike syndrome. All of these patients have accessory limbs, and 2 of them have accessory genitourinary structures. Interestingly, one of these patients has extrophia vesicalis of accessory bladder. None of them have chromosomal abnormality. Here the authors present distribution of findings of these cases. PMID- 15359418 TI - A case of bladder exstrophy and thoracal intramedullary teratoma. AB - Exstrophic bladder and intramedullary teratomas are rare congenital anomalies. To the authors' knowledge, the coexistence of these 2 anomalies has not been reported previously. The authors report on a newborn with thoracal intramedullary teratoma and exstrophic bladder. The possible embryogenetic background also is discussed. PMID- 15359419 TI - Patternhunter II: highly sensitive and fast homology search. AB - Extending the single optimized spaced seed of PatternHunter(20) to multiple ones, PatternHunter II simultaneously remedies the lack of sensitivity of Blastn and the lack of speed of Smith-Waterman, for homology search. At Blastn speed, PatternHunter II approaches Smith-Waterman sensitivity, bringing homology search methodology research back to a full circle. PMID- 15359420 TI - Reconstructing genetic networks from time ordered gene expression data using Bayesian method with global search algorithm. AB - Different genes of an organism are expressed to different levels at different times during the life cycle and in response to various environmental stresses. Elucidating the network of gene-gene interactions responsible for the expression helps understand living processes. Microarray technology allows concurrent genomic scale measurement of an organism's mRNA levels. We describe a power-law formalism to model the combinatorial effect of regulators on gene transcription. The dynamic model allows delayed transcription. We employ a principled network reconstruction approach that accounts for the high noise and low replicate characteristics of present day microarray data. An important feature of our approach is that the detail of the reconstructed network is limited to the noise level of the data. We apply the methodology to a microarray dataset of yeast cells grown in glucose and experiencing a diauxic transition upon glucose depletion. The reconstructed transcriptional regulations of yeast glycolytic genes are consistent with published findings. PMID- 15359421 TI - Supersequences of masks for oligo-chips. AB - On a very small surface, a chip, several thousands of oligonucleotides, can be synthesized using a mask technology. Given a set of oligos, the problems tackled here are: What is the minimum number of masks necessary to synthesize one copy of each oligo? How long will be the series of masks if each oligo is synthesized twice, such that the two copies are realized with two completely different series of masks? We establish that, for 20,000 oligos of 20 bases a single copy can be synthesized with around 67 masks and for two copies with less than the double. PMID- 15359422 TI - CooPPS: a system for the cooperative prediction of protein structures. AB - Predicting the three-dimensional structure of proteins is a difficult task. In the last few years several approaches have been proposed for performing this task taking into account different protein chemical and physical properties. As a result, a growing number of protein structure prediction tools is becoming available, some of them specialized to work on either some aspects of the predictions or on some categories of proteins; however, they are still not sufficiently accurate and reliable for predicting all kinds of proteins. In this context, it is useful to jointly apply different prediction tools and combine their results in order to improve the quality of the predictions. However, several problems have to be solved in order to make this a viable possibility. In this paper a framework and a tool is proposed which allows: (i) definition of a common reference applicative domain for different prediction tools; (ii) characterization of prediction tools through evaluating some quality parameters; (iii) characterization of the performances of a team of predictors jointly applied over a prediction problem; (iv) the singling out of the best team for a prediction problem; and (v) the integration of predictor results in the team in order to obtain a unique prediction. A system implementing the various steps of the proposed framework (CooPPS) has been developed and several experiments for testing the effectiveness of the proposed approach have been carried out. PMID- 15359423 TI - Protein classification using comparative molecular interaction profile analysis system. AB - We recently introduced a new molecular description factor, interaction profile Factor (IPF) that is useful for evaluating molecular interactions. IPF is a data set of interaction energies calculated by the Comparative Molecular Interaction Profile Analysis system (CoMIPA). CoMIPA utilizes AutoDock 3.0 docking program, and the system has shown to be a powerful tool in clustering the interacting properties between small molecules and proteins. In this report, we describe the application of CoMIPA for protein clustering. A sample set of 15 proteins that share less than 20% homology and have no common functional motifs in primary structure were chosen. Using CoMIPA, we were able to cluster proteins that bound to the same small molecule. Other structural homology-based clustering programs such as PSI-BLAST or PFAM were unable to achieve the same classification. The results are striking because it is difficult to find any common features in the active sites of these proteins that share the same ligand. CoMIPA adds new dimensions for protein classification and has the potential to be a helpful tool in predicting and analyzing molecular interactions. PMID- 15359424 TI - Reduced bio-basis function neural networks for protease cleavage site prediction. AB - This paper presents a new neural learning algorithm for protease cleavage site prediction. The basic idea is to replace the radial basis function used in radial basis function neural networks by a so-called bio-basis function using amino acid similarity matrices. Mutual information is used to select bio-bases and a corresponding selection algorithm is developed. The algorithm has been applied to the prediction of HIV and Hepatitis C virus protease cleavage sites in proteins with success. PMID- 15359425 TI - An integrated comprehensive workbench for inferring genetic networks: voyagene. AB - We propose an integrated, comprehensive network-inferring system for genetic interactions, named VoyaGene, which can analyze experimentally observed expression profiles by using and combining the following five independent inferring models: Clustering, Threshold-Test, Bayesian, multi-level digraph and S system models. Since VoyaGene also has effective tools for visualizing the inferred results, researchers may evaluate the combination of appropriate inferring models, and can construct a genetic network to an accuracy that is beyond the reach of a single inferring model. Through the use of VoyaGene, the present study demonstrates the effectiveness of combining different inferring models. PMID- 15359426 TI - Bioie: retargetable information extraction and ontological annotation of biological interactions from the literature. AB - The need for extracting general biological interactions of arbitrary types from the rapidly growing volume of the biomedical literature is drawing increased attention, while the need for this much diversity also requires both a robust treatment of complex linguistic phenomena and a method to consistently characterize the results. We present a biomedical information extraction system, BioIE, to address both of these needs by utilizing a full-fledged English grammar formalism, or a combinatory categorial grammar, and by annotating the results with the terms of Gene Ontology, which provides a common and controlled vocabulary. BioIE deals with complex linguistic phenomena such as coordination, relative structures, acronyms, appositive structures, and anaphoric expressions. In order to deal with real-world syntactic variations of ontological terms, BioIE utilizes the syntactic dependencies between words in sentences as well, based on the observation that the component words in an ontological term usually appear in a sentence with known patterns of syntactic dependencies. PMID- 15359427 TI - Identifying patterns of DNA for tumor diagnosis using capillary electrophoresis amplified fragment length polymorphism (CE-AFLP) screening. AB - Amplified Fragment Length Polymorphism (AFLP) screening is a genome-wide genotyping strategy that has been widely used in plants and bacteria, but little has been reported concerning its use in humans. We investigated if the AFLP procedure could be coupled with high-throughput capillary electrophoresis (CE) for use in tumor diagnosis and classification. Using CE-AFLP, a series of molecular 'fingerprints' were generated for a set of gastric tumor and normal genomic DNA samples. The CE-AFLP procedure was qualitatively and quantitatively robust, and a variety of clustering tools were used to identify a specific DNA marker 'pattern' of 20 features that classified the tumor and normal samples to reasonable degrees of accuracy (Sensitivity 95%, Specificity 80%). The CE-AFLP based approach also correctly classified 16 tumor samples, which in a previous study had exhibited no detectable genomic aberrations by comparative genome hybridization (CGH). This is the first reported application of CE-AFLP screening in tumor diagnosis. As the procedure is relatively inexpensive and requires minimal prior sequence knowledge and biological material, we suggest that CE-AFLP based protocols may represent a promising new approach for DNA-based cancer screening and diagnosis. PMID- 15359428 TI - Robustness of metabolic map reconstruction. AB - With the ever increasing amount of genomic data available, the interest for generating biochemical pathways has grown tremendously. So far, mainly complete genomes have been used to reconstruct the biochemical pathways and their associated interactions. However, a large number of low coverage genomes, as well as other sources of partial genomic data, are currently available for many organisms. In order to be able to use incomplete data for metabolic reconstruction, the inherent properties of this procedure need to be investigated. In this short note, we describe the robustness and predictive power of metabolic reconstructions using partial information from Schizosaccharomyces pombe. We also discuss the implications of the results on reference genome projects as well as other large-scale sequencing data. PMID- 15359429 TI - Diastolic heart failure. PMID- 15359430 TI - Diastolic heart failure. PMID- 15359431 TI - Obesity and the metabolic syndrome in children and adolescents. PMID- 15359432 TI - Obesity and the metabolic syndrome in children and adolescents. PMID- 15359433 TI - Palliative care. PMID- 15359434 TI - Case 11-2004: a boy with rash, edema, and hypertension. PMID- 15359435 TI - [Diverticulitis of the gall bladder]. PMID- 15359436 TI - Khousalya Periaswamy. Interview by Bob Huff. PMID- 15359437 TI - Keystone HIV pathogenesis and vaccine development report. PMID- 15359438 TI - Abbott's Norvir price hike is bad medicine. Statement at the NIH public meeting on Norvir. PMID- 15359439 TI - FDA issues warning letter to Abbott over Norvir pricing spin. PMID- 15359440 TI - Drug news. PMID- 15359441 TI - Scramble for Africa. PMID- 15359442 TI - Partners in care. Dr. William Johnson and Bethsheba Johnson continue the vision of Dr. Sherry Luck. PMID- 15359443 TI - Who are we hurting? The hesitation by people of color. PMID- 15359444 TI - Vaccines--a community role for HIV. The time to get involved is now. PMID- 15359445 TI - Children's research--a mom's experience. One family's struggle to survive. PMID- 15359446 TI - Going to extremes to stay alive. From study to study until the cure. PMID- 15359447 TI - Who's afraid of the big bad study? Trials to think about. PMID- 15359448 TI - Crystal methamphetamine and HIV--a catastrophe. PMID- 15359449 TI - Community outreach. Community efforts supported in Africa. PMID- 15359450 TI - Experts: AIDS should rival terrorism for world spotlight, action. PMID- 15359451 TI - HIV and gender. Conference calls for response to specific needs of women. PMID- 15359452 TI - Employment discrimination. HIV-positive man who quit his job loses bias claim. PMID- 15359453 TI - Insurance benefits reinstated after HIV diagnoses. PMID- 15359454 TI - Antiretrovirals. WHO misses 6-month target for distributing AIDS drugs. PMID- 15359455 TI - Justice department finds HIV down, AIDS up in prisons. PMID- 15359457 TI - Juvenile testing. Order for forced HIV testing of youth reversed on appeal. PMID- 15359456 TI - Alien with HIV can review guilty plea before deportation. PMID- 15359458 TI - Breach of confidentiality actions to proceed against doctors. PMID- 15359459 TI - Court-ordered HIV testing upheld for sex offender. PMID- 15359460 TI - Discrimination. Casino cleared of bias over firing of HIV-positive employee. PMID- 15359462 TI - Politics. Researchers say Bush administration suppresses science. PMID- 15359461 TI - Sexual offender to undergo involuntary HIV testing. PMID- 15359463 TI - [Psychiatric training of nurses in the Netherlands since 1883]. AB - This article describes the development since 1883 of training nurses for working with psychiatric patients. Knowledge of this development seems to throw a new light upon the nursing history in the Netherlands. To start with, the vocational training system in psychiatric hospitals developed during exactly the same time as the hospital based educational system for so called ' general nurses'. As a result, both these vocational training routes were acknowledged as 'basic learning routes for nurses' in the Dutch Nursing Law of 1921. The general route gave to Diploma A, the psychiatric route gave access to Diploma B. This double diploma system placed the Dutch training system for nurses in an isolated position. For most other countries - as well as the International Council of Nurses - considered psychiatry as a field of work in which a nurse could specialise only after having completed a basic training programme as a general nurse. In most countries, the specialization course in psychiatry only took one year. In comparison to those systems, the Dutch programme seemed to offer psychiatric nurses more years of specific training. After World War II, however, also in the Netherlands many people advocated a one diploma system in nursing, with possibilities to specialise. Nevertheless, it took the Dutch until 1997 to introduce this new system, so it seems the old training routes were defended vehemently. Maybe an historic description of the way this psychiatric route had developed, can shed new light upon the aspects of the old educational system that one longed to preserve. PMID- 15359464 TI - [The role of papyri in the history of medicine]. AB - Two papyri are very interesting for Medicine history of the Antiquity. The first one (a Greek papyrus prolifically illustrated between 100 B.C. and 100 A.D.) reveals the value of the illustrators of works in the Greek and Roman period and the second (the famous papyrus of Milan 300 B.C.) includes a part about the epigrams of curing. PMID- 15359465 TI - Incidence of tuberculosis in Olmsted County, Minnesota, 1990-2001. AB - OBJECTIVE: To describe and compare the incidence and clinical characteristics of tuberculosis in Olmsted County, Minnesota, among US-born and foreign-born persons. PATIENTS AND METHODS: We performed a retrospective cohort study at the Mayo Clinic in Rochester, Minn, of all residents of Olmsted County (2000 population: 124,277) diagnosed as having tuberculosis between January 1, 1990, and December 31, 2001. Potential cases were identified with use of a computerized diagnostic coding database and microbiological laboratory data; all identified medical records were abstracted. Definite cases were those in which Mycobacterium tuberculosis was recovered in culture. Probable cases were those that met predefined clinical or radiographic evidence of tuberculosis and other criteria. Age-specific, sex-specific, and country of origin-specific incidence rates were calculated with use of Olmsted County census data. Variables were compared among risk groups using the Fisher exact test. RESULTS: During a 12-year period, 71 cases of tuberculosis (53 definite, 18 probable) were identified, for an incidence of 5.3 per 100,000 person-years. Of these cases, 54 (76%) occurred during the second half of the study (incidence: 7.7 per 100,000 person-years). The incidence among US-born persons was similar throughout the study period; however, the Incidence among foreign-born persons increased more than 3-fold during the second half of the study period. Twenty-five patients (35%) were former refugees. All isoniazid-resistant infections (12% of isolates) and multidrug-resistant infections (6% of isolates) occurred among foreign-born persons. CONCLUSION: The incidence of tuberculosis increased substantially in Olmsted County between 1990 and 2001, primarily because of an increase in the number of cases among foreign-born persons. PMID- 15359466 TI - Toxoplasma gondii infections in cats from Parana, Brazil: seroprevalence, tissue distribution, and biologic and genetic characterization of isolates. AB - Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that can excrete environmentally resistant oocysts. The prevalence of T. gondii was determined in 58 domestic cats from 51 homes from Santa Isabel do Ivai, Parana State, Brazil where a water-associated outbreak of acute toxoplasmosis had occurred in humans. Antibodies to T. gondii were found with the modified agglutination test in 49 of 58 (84.4%) cats at a serum dilution of 1:20. Tissues (brain, heart, and skeletal muscle) of 54 of these cats were bioassayed in T. gondii-free, laboratory-reared cats; T. gondii oocysts were excreted by 33 cats that were fed feline tissues. Brains from these 54 cats were bioassayed in mice; T. gondii was isolated from 7. Skeletal muscles and hearts of 15 cats were also bioassayed in mice; T. gondii was isolated from skeletal muscles of 9 and hearts of 13. The results indicate that T. gondii localizes in muscle tissue more than the brains of cats. In total there were 37 T. gondii isolates from 54 cats. Most isolates of T. gondii were virulent for mice. Genotyping of the 37 isolates of T. gondii, using the SAG2 locus, revealed that 15 isolates were type I and 22 were type III. The absence of type II genotype in cats in this study is consistent with the previous studies on T. gondii isolates from Brazil and is noteworthy because most T. gondii isolates from the United States are type II. These findings support the view that Brazilian and North American T. gondii isolates are genetically distinct. This is the first report of genotyping of T. gondii isolates from the domestic cat. PMID- 15359467 TI - Mouse manipulation through single-switch scanning. AB - Given the current extensive reliance on the graphical user interface, independent access to computer software requires that users be able to manipulate a pointing device of some type (e.g., mouse, trackball) or be able to emulate a mouse by some other means (e.g., scanning). The purpose of the present study was to identify one or more optimal single-switch scanning mouse emulation strategies. Four alternative scanning strategies (continuous Cartesian, discrete Cartesian, rotational, and hybrid quadrant/continuous Cartesian) were selected for testing based on current market availability as well as on theoretical considerations of their potential speed and accuracy. Each strategy was evaluated using a repeated measures study design by means of a test program that permitted mouse emulation via any one of four scanning strategies in a motivating environment; response speed and accuracy could be automatically recorded and considered in view of the motor, cognitive, and perceptual demands of each scanning strategy. Ten individuals whose disabilities required them to operate a computer via single switch scanning participated in the study. Results indicated that Cartesian scanning was the preferred and most effective scanning strategy. There were no significant differences between results from the Continuous Cartesian and Discrete Cartesian scanning strategies. Rotational scanning was quite slow with respect to the other strategies, although it was equally accurate. Hybrid Quadrant scanning improved access time but at the cost of fewer correct selections. These results demonstrated the importance of testing and comparing alternate single-switch scanning strategies. PMID- 15359468 TI - Enzymatic reactions involved in the repair of oxidized proteins. AB - Proteins are the targets of reactive oxygen species, and cell aging is characterized by a build-up of oxidized proteins. Oxidized proteins tend to accumulate with age, due to either an increase in the rate of protein oxidation, a decrease in the rate of oxidized protein repair and degradation, or a combination of both mechanisms. Oxidized protein degradation is mainly carried out by the proteasomal system, which is the main intracellular proteolytic pathway involved in protein turnover and the elimination of damaged proteins. However, part of the oxidative damage to cysteine and methionine residues, two amino acids which are highly susceptible to oxidation, can be repaired by various enzymatic systems that catalyze the reduction of cysteine disulfide bridge, cysteine-sulfenic and -sulfinic acids as well as methionine sulfoxide. The aim of this review is to describe these enzymatic oxidized protein repair systems and their potential involvement in the decline of protein maintenance associated with aging, focusing in particular on the methionine sulfoxide reductases system. PMID- 15359469 TI - Transvenous biopsy of cavo-atrial tumors with the Quick-Core needle. AB - A variant application of the renal Quick-Core needle in 3 patients with cavo atrial tumors is reported. In all 3 patients either a transjugular or transfemoral venous biopsy approach with this device yielded sufficient tissue for histological diagnosis at the first attempt. Bioptic diagnoses were confirmed either by surgery or radiological and clinical response to a specific chemotherapy. There were no procedure-related complications. PMID- 15359470 TI - Treatment of acute lower limb ischemia following the use of the Duett sealing device: report of three cases and review of the literature. AB - Three cases of local thrombolysis in the treatment of acute lower limb ischemia complicating the utilization of the Duett sealing device are presented. Routine usage of several vascular closure devices after cardiac catheterization and percutaneous coronary intervention (PCI) has been adopted in our institution during the last 3 years (September 1999 to April 2003). The Duett closure device has been used in 420 patients (post-coronary angiography, 359; post-PCI, 61). Three patients (0.7%) demonstrated acute leg ischemia used by inadvertent intravascular administration of the sealing material related to this device. All three were treated successfully by catheter-directed local thromolysis (tissue plasminogen activator 5 mg bolus followed initially by 1 mg/hr and consequently by 0.5-1.0 mg/hr depending upon the development of significant hematoma and lasting for 24 hr). In conclusion, interventional treatment using local thrombolysis should be the first-line treatment in acute lower limb ischemia complicating the utilization of the Duett sealing device. PMID- 15359471 TI - Successful covering of a hepatic artery aneurysm with a coronary stent graft. AB - In a 54-year-old woman with liver cirrhosis who underwent orthotopic liver transplantation, the postoperative course was complicated by aneurysm formation in the hepatic artery. Abdominal ultrasonography showed a daily increase in size of the aneurysm in spite of careful management including strict rest and continuous intravenous infusion of antihypertensive agents. Since the patient's poor systemic status was a major obstruction to operative resection, transcatheter therapy was though more preferable. We evaluated the lesion with intravascular ultrasonography as an adjunct to angiography and dissection with a flap was well visualized. The aneurysm was covered with a commercially available stent-graft, designed for treatment of the coronary artery. This is a rare case in which a Jostent was implanted into the hepatic artery after liver transplantation. PMID- 15359472 TI - CO2 demonstration of multiple extravasations into a subcapsular hematoma of the liver. AB - In a case of esophageal cancer with liver metastases, rupture of a liver metastasis resulted in subcapsular hematoma of the liver. Digital subtraction angiography with carbon dioxide showed multiple extravasations at the surface of the liver suggesting multiple ruptures of the penetrating hepatic capsular arteries. It was suggested that these findings are not rare in cases of subcapsular hematoma; however, they have received little attention. PMID- 15359473 TI - Percutaneous sclerotherapy using acetic acid after failure of alcohol ablation in an intra-abdominal lymphangioma. AB - We report a case of percutaneous sclerotherapy using acetic acid in a 22-year-old woman with intra-abdominal cystic lymphangioma who was not successfully treated with ethanol despite multiple trials. PMID- 15359474 TI - A case of aorto-bronchial fistula after insertion of left main bronchial self expanding metallic stent in a patient with recurrent esophageal cancer. AB - We report a case of aorto-bronchial fistula (ABF) caused by a self expanding metallic stent (EMS) 51 days after insertion into the left main bronchus. The patient presented with left main bronchial stenosis caused by post-operative local recurrence of esophageal cancer. Post-operative radio therapy totaling 40 Gy and post-recurrence radiotherapy totaling 34 Gy were administered, with daily fractions of 2 Gy. Stenosis of the left main bronchus improved slightly, and was followed with insertion of EMS to prevent restenosis. The patient experienced massive hemoptysis for 3 days before sudden death. Autopsy revealed the EMS edge perforating the descending aortic lumen. Tumor infiltration and bacterial infection were observed on the wall of the left bronchus, and atherosclerosis was present on the aortic wall around the fistula. It should be noted that the left main bronchus was at considerable risk of ABF after insertion of EMS for malignant stenosis, and prophylactic sent insertion into the left bronchus without imperative need must be avoided. PMID- 15359475 TI - Subperiosteal orbital hemorrhage complicating cardiac surgery. AB - Subperiosteal orbital hemorrhage (SPOH) following cardiac surgery has not been previously reported. We present a patient who developed diplopia and right eye proptosis immediately after cardiac surgery for a mitral valve repair and coronary artery bypass graft. A computed tomography (CT) study demonstrated a right superior SPOH. The diplopia and proptosis resolved spontaneously within 4 weeks. Follow-up CT showed complete resolution of the SPOH. PMID- 15359476 TI - Peripheral MR angiography of Klippel-Trenaunay syndrome. PMID- 15359477 TI - [Bacteriological war (1916-1933): from anecdote to terror]. AB - Rotten corpses were occasionally used as weapons in the Antiquity. So wells and springs were usually contaminated. In 1915-1916, just after the use of poison gas on the battlefield, bacteriological suspensions were prepared to destroy men and cattle in Europe, Russia and the United of America but the attempts of using bacteriological weapons remained inefficient. Moreover the evidences of this use during the Great War are very fragile. After the war there was a consensus about inhumanity of this weapon which yet might be used if the enemy did it. Between 1925 and 1933 the debates in the Society of Nations proved that bacteriological war was possible in spite of a compromise solution between some nations. Then English journalist Wickdam Steed disclosed the German tries of bacteriological weapons in the Paris metro and the London tube in 1933 (Was this article a manipulation?). PMID- 15359478 TI - [The voyage in 1547 to Stamboul (Constantinople) by the pharmacist-naturalist Pierre Belon from Mans]. AB - Pierre Belon was born in Le Mans in 1517. After he had worked as apothecary assistant and he had studied botany, he was chosen to join in the embassy sent by French King Francois I to the Turkish Sultan in 1546. From 1553 he wrote his impressions upon his tour "Comments upon different peculiarities and memorable things". In Stamboul he dawdled around the typical Turkish drugstores and he observed and recorded the hygienic habit of the Turks. Not only are his "Comments" about the Eastern Countries a mine of useful information but they can be also considered as the foundation of renewal in the relationship between Europeans travellers and the Middle East. PMID- 15359479 TI - [The diagnostic contributions and pharmaceutic prescriptions of the Geneva physician Theodore Tronchin in 1763]. AB - The Geneva physician Theodore Tronchin (1709-1801) was one of the best known physicians in Europe. Some copies of his letters giving medical advice to his patients all over the world are preserved in the University Library of Geneva. One of them dated 1763 was chosen as an example of Tronchin's dealings with his patients. In it Tronchin gives his opinion about the venereal disease transmitted to princess Marie-Therese of Hohenzollern-Hechingen (1732-1802) by her husband. Tronchin prescribed treatment based on mercury and sudorific plants. PMID- 15359480 TI - [Medical practice and the feminine sense of modesty in the 19th century]. AB - The feminine sense of modesty appeared as one of the main difficulties that the physicians of the 19th century met to examine the sex of a female patient. In order to convince of a perfect examination, numerous physicians strove to respect the sense of modesty during the consultation and would avoid to expose the whole body. Nevertheless some emphasised that taking precautions could not lead to neglect the consultation. PMID- 15359481 TI - Rest and restraint. PMID- 15359482 TI - A happy ending? PMID- 15359483 TI - [The pewter bleeding bowls]. AB - In the late seventeenth century, then along the eighteen and nineteenth centuries the amount of the bloodlettings was measured by means of three pewter bleeding bowls that held three ounces of blood, about 300 millilitres. In the middle of the nineteenth century new and large bleeding bowls with metric graduations were manufactured only by the Parisian potters. PMID- 15359484 TI - Man vs. insect. PMID- 15359485 TI - The motives behind Cantor's set theory - physical, biological, and philosophical questions. PMID- 15359486 TI - Proceedings of the International Symposium on the Pathogenesis of SARS. July 12 13, 2003, Beijing, China. PMID- 15359487 TI - Literature alert. Endothelial dilatory function predicts individual susceptibility to renal damage in the 5/6 nephrectomized rats. PMID- 15359488 TI - Obesity: the public health challenge of our time. PMID- 15359489 TI - Role of endothelin ETB receptor in the pathogenesis of monocrotaline-induced pulmonary hypertension in rats. AB - We investigated the role of endothelin ETB receptor in the development of monocrotaline-induced pulmonary hypertension, by using the spotting-lethal (sl) rat, which carries a naturally occurring deletion in the endothelin ETB receptor gene. Three weeks after injection of saline or monocrotaline (60 mg/kg, s.c.), hemodynamics, cardiac hypertrophy and endothelin-1 levels in right ventricle were determined. Monocrotaline produced a marked pulmonary hypertension associated with increases in right ventricular pressure and hypertrophy, pulmonary arterial medial thickening and the endothelin-1 levels. The monocrotaline-induced alterations tended to be enhanced in ETB-deficient homozygous rats, compared with cases in wild-type rats. The treatment with selective ETA receptor antagonist ABT 627 [2R-(4-methoxyphenyl)-4S-(1,3-benzodioxol-5-yl)-1-(N,N-di(n butyl)aminocarbonyl-methyl)-pyrrolidine-3R-carboxylic acid] for 3 weeks (10 mg/kg/day, twice daily) almost completely suppressed the monocrotaline-induced pulmonary hypertension and related organ damage both in ETB-deficient and wild type animals to the same levels. Thus, we suggest that the antagonism of the ETA receptor is essential for the protection from monocrotaline-induced pulmonary hypertension, irrespective of the presence of the ETB receptors, although a protective role of ETB receptor-mediated action in the pathogenesis of this disease model cannot be ruled out. PMID- 15359490 TI - [The risk factors of childhood injuries and their intervention and control]. PMID- 15359491 TI - [Diagnosis and treatment of primary hyperaldosteronism]. PMID- 15359492 TI - [Endoscopy in plastic and reconstructive surgery]. PMID- 15359493 TI - [Contemporary concept of chronic stomach ulcer as precancerous state]. PMID- 15359495 TI - Lysle E. Johnston, Jr: a perspective. PMID- 15359496 TI - Taking stock: assessing the present, planning the future. PMID- 15359497 TI - Taking stock: passing the torch, who has the light? Reflections about teaching orthodontics at the occasion of the retirement of Prof. Lysle E. Johnston, Jr PhD. AB - In the Marburg (Germany) 5-year dental curriculum, students enter the orthodontic department in the middle of their third year. Our teaching is student-centered and problem-solving oriented. From day 1 our students are trained to diagnose in three separate levels of development i.e. (i) craniofacial growth, (ii) development of the dentition and (iii) maturation of function. These three levels are kept separate throughout patient examination and up to the strategic treatment planning phase. The examination is performed with the help of three checklists, one for the facial morphology, one for the dentition and one for function. The three resulting problem lists then undergo reduction by selecting one 'key problem' for each level. Next, for every key problem three 'standard solutions' are offered, not in the form of appliances but as vectors that can modify growth. Finally, in a strategic planning phase, one treatment option from each of the three levels is selected and combined in a single, be it hypothetical, treatment plan. Parallel to these, students are exposed to treatment mechanics through the presentation of selected orthodontically treated patients. In our experience the advantage of this diagnostic procedure lies in the structured organization that serves as a GPS system for student and teacher and enables them a clear communication about where they are and what has to be done. By design, treatment options have taken priority over treatment mechanics. We are confident that our students, equipped with real life diagnostic skills, are well prepared for their orthodontic future. PMID- 15359498 TI - Taking stock: a century of orthodontics--has excellence been redefined as expediency? PMID- 15359499 TI - Taking stock: assessing the present-planning the future 'you don't need a weatherman to know which way the wind blows'. AB - In the past, the dual nature of orthodontics has presented practitioners with unique artistic challenges and scientific opportunities. As a result, orthodontists today are enjoying the 'Platinum Age of Productivity.' However, a disturbing trend that promotes neither art nor science has emerged. Health care reform in the United States has transformed health care from a service to a commodity. Some orthodontists and orthodontic practice management companies have capitalized on this change in thinking and are aggressively promoting orthodontic treatment as a product. This paper examines the impact of this paradigm shift on the future of orthodontics. Particular attention is paid to the interaction of three themes: first, orthodontics as a product, secondly, the current misconception that there is a shortage of orthodontic practitioners in the United States, and thirdly, the difficulty in defining 'excellence' in specialty training in orthodontics. In the near future, the collision of these three concepts will result in an orthodontic 'Perfect Storm'. The ability of our specialty to survive and thrive will be proportional to our professional behavior during these difficult times...caveat emptor. PMID- 15359500 TI - Taking stock: a critical perspective on contemporary orthodontics. AB - This essay briefly considers some underlying concerns about the direction of clinical research in orthodontics. The field is viewed from a personal perspective as things are now, as they are becoming, and as I believe they should be. PMID- 15359502 TI - Taking stock: hippocratic and platonic thoughts on orthodontic tooth movement. PMID- 15359501 TI - Taking stock: from chasing occlusal contacts to vulnerability alleles. AB - Over the past 70 years, temporomandibular disorders (TMDs) have been subject to shifts in conceptual understanding. Unable to account for disease patterns, the mismatch between case assignment and treatment need, and very different interventions producing similar treatment outcomes (except for the risk to patients), emerging theories make persuasive arguments in support of alternative explanations. PMID- 15359503 TI - Bone structure effect on root resorption. AB - OBJECTIVES: To explore if alveolar bone shape and density might promote external apical root resorption. SETTING AND SAMPLE POPULATION: Panoramic radiographs of 700 patients who had orthodontic treatment at Temple University were reviewed and 22 patients with radiographic evidence of root resorption on the lower incisors were selected for the study. Exclusion criteria included a history of systemic diseases, craniofacial abnormalities, tooth injury, endodontically treated teeth, and impacted teeth. METHODS: Pre-treatment (T1) and post-treatment (T2) cephalometric radiographs were converted into digital format and enhanced to reduce contrast variability and improve edge definition. Tooth length, root length, root area, alveolar area around the root including cortical area, area of medullary bone, and area of the symphysis were measured using an interactive software algorithm. A region of interest within the symphysis was also defined and trabecular space area and fractal dimension calculated as an estimate of bone density. RESULTS: Root area and tooth length were correlated negatively with changes in root area, tooth area, and root length. Larger teeth demonstrated a greater amount of root resorption. Dentoalveolar complex dimensions remained relatively unchanged during tooth movement. The amount of alveolar bone around the root, thickness of cortical bone, density of the trabecular network, and fractal dimension showed no significant correlation with the extent of the external apical root resorption. CONCLUSIONS: The results of this study suggest that the density and morphology of the dentoalveolar complex are not significant factors in the etiology of external apical root resorption. PMID- 15359504 TI - Ehlers-Danlos Syndrome (EDS) focusing on oral symptoms: a questionnaire study. AB - OBJECTIVES: To study the prevalence of oral problems reported among a large group of adults with Ehlers-Danlos Syndrome (EDS). Furthermore, to compare proportions of disorders and general psychological well-being with those in a cohort of randomized population-based controls. DESIGN: A questionnaire study. SETTING AND SAMPLE POPULATION: A total of 265 persons with EDS (response rate 77%) and 750 controls (response rate 63%) in a population-based cohort. Sixty persons were excluded from the EDS group (support members, children, undiagnosed EDS) leaving a final study group of 144 adults. The final control group consisted of 331 persons since many had sent back unanswered questionnaires. EXPERIMENTAL VARIABLES: Questions concerning general and oral symptoms, VAS scales for masticatory muscle pain ratings and General Health Questionnaire (GHQ12) questions were included. OUTCOME MEASURE: Proportions of affirmative answers in EDS group and controls were compared. Mean values for ratings on VAS scales. Proportions of GHQ12 scores with a threshold score of four or more. RESULTS: The proportions of affirmative answers for persons with EDS concerning general health problems, oral problems and masticatory muscle symptoms were significantly higher compared with controls. In the EDS group those who had daily pain in the masticatory muscles had significantly higher mean values on the VAS scales compared with those who had pain a few days per month. A GHQ12 score of four or more was significantly more common in the EDS group than in the controls, suggesting that a decline in psychological well-being was also more common in this group. CONCLUSION: It is important that dental practitioners should be aware of the oral problems associated with EDS and the impact the disease has on quality of life. PMID- 15359505 TI - Effects of the anticholinesterase drug tacrine on the development of PTZ kindling and on learning and memory processes in mice. AB - INTRODUCTION: The anticholinesterase drug tacrine (Tac) is known to have a beneficial effect on memory deficit in mice involving in the process the cholinergic system in the brain. Chemically induced kindling is a well-known model of petit mal epilepsy. Kindling develops after repeated administration of subconvulsant doses of pentylenetetrazole (PTZ). This model is suitable for studying the two CNS disturbances associated with epilepsy, i.e. seizure attacks and memory impairment. OBJECTIVE. The aim of the present study was to examine the effect of the anticholinesterase drug Tac on the model of PTZ kindling and on active avoidance in mice. METHODS. Kindling was induced by repeated administration of PTZ (40 mg/kg) s.c. at 48-hour intervals. Tac (at doses of 0.1, 0.5 and 1 mg/kg) was injected 40 minutes before PTZ over an 8 - week period in 4 experimental groups of mice. Seizure intensity was scored using a 5-grade scale. Kindling was defined as 3, 4 or 5 grade seizures evoked by 3 consecutive doses of PTZ and treatment was discontinued. The active avoidance method (training session consisting of 50 trials) was used to test learning and memory functions. On day 7 following the learning session a memory retention test was performed. The challenge dose of PTZ was given after a 15-day discontinuation of treatment. RESULTS: In control mice, kindling was developed over a 7-week period of treatment. Mice injected with Tac (in 3 consecutive doses) showed increased seizure intensity from week 1 through week 7 of the study. On learning and memory retention tests kindled mice showed a reduced learning capabilities whereas a statistically significant upward tendency in the number of conditioned responses (avoidances) was recorded in mice treated with Tac (1 mg/kg), when compared with the controls. Mice treated with Tac (in doses of 0.1 and 0.5 mg/kg) also exhibited an increased number of avoidances on learning and memory retention tests. CONCLUSIONS: 1. The anticholinesterase drug Tac increased the seizure intensity and facilitated the development of PTZ kindling in mice. 2. On the basis of well-developed kindling Tac did not impair the memory of the experimental animals, which is possibly due to its CNS stimulating effect. PMID- 15359506 TI - A fifteen-month survey of directly observed therapy--short course and antibiotic drug resistance in the region of Plovdiv. AB - OBJECTIVE: This study addresses the issue of antituberculosis drug resistance in a cohort of 213 patients from Plovdiv included in the pilot phase of a DOTS based project for a 15-month period. Between July 1. 1998 and September 30, 1999, ninety three culture-positive patients participated in the study. 89 of them were tested for drug susceptibility to rifampicin, izoniazid, etambutol, and streptomycin. RESULTS: Resistance to at least one antituberculosis drug was established in 24.7% of the patients. Monoresistance was found in 13.5% of the cases. The median prevalence of combined resistance to rifampicin and isoniazid was 6.7%. The prevalence of resistance to rifampicin or isoniazid was 21.4%. Drug susceptibility testing results were obtained within 67 days. In 33% of the patients continuation treatment phase was initiated before drug susceptibility data were available. CONCLUSIONS: During the observed period a considerably high rate of drug resistant tuberculosis was registered among the patients included in the pilot phase of the program based on Directly Observed Therapy--Short Course. The percentage of resistance to R and/or I gives better information about the risk of inadequate treatment during the continuation treatment phase. The high percentage of this pattern of resistance in our region requires the continued use of four first-line drugs for therapy until the results of drug-susceptibility testing are received. PMID- 15359507 TI - Arthroscopic evaluation of chondropathy in knee osteoarthritis. AB - Knee arthroscopy under local anesthesia is an invasive, highly informative diagnostic method. It allows the gross evaluation of pathological changes in joint cartilage in knee osteoarthritis. The evaluation of chondropathy is usually performed using a four-degree scale or using a visual analog scale. The main characteristics of arthroscopy can be summarized as follows: simplicity, high intrinsic validity, good extrinsic validity, intra- and interobserver reliability. Arthroscopy has a high degree of clinical relevance and sensitivity to pathological changes. PMID- 15359508 TI - Relevance between arthroscopic pathology and clinical characteristics in knee osteoarthritis. AB - Arthroscopy can be used to identify accurately cartilage and synovial changes in knee osteoarthritis (OA). The objective of this study was to find the correlation between arthroscopically established knee chondropathy and synovitis and the corresponding algofunctional characteristics in OA. METHODS: 41 patients with OA in only one knee joint were investigated. The diagnosis of knee OA fulfilled the clinical, laboratory and radiological ARA criteria. Low pressure arthroscopy of the affected knee was performed under local anaesthesia. Chondropathy was evaluated according to the method of R E Outerbridge. Synovial inflammation was assessed by the scoring system of S. Lindblad and E. Hedfors. RESULTS: We found statistically significant correlation (Spearman's rho test) between chondropathy of the knee and the following clinical indices: pain at night, pain after standing 30 minutes, pain on walking, pain on getting up from a chair without help of arms, maximum walking distance. We failed to demonstrate statistical significant correlation between chondropathy and morning stiffness and activities of daily living. There was also no statistical correlation between knee synovitis and any of the Lequesne's clinical indices. CONCLUSION: The arthroscopically established scores of the cartilage pathology were consistent with most of the clinical algofunctional symptoms. The degree of synovitis in our study was not correlated with clinical activity symptoms of OA, according to Leguesne. Five positive algofunctional Lequesne's scores corresponded to heavy breakdown of knee cartilage in OA. PMID- 15359509 TI - Ultrasonographic evaluation of knee joint cartilage in rheumatoid arthritis patients. AB - The non-invasive methods used to study joint cartilage are restricted in their scope. No direct visualization of the joint cartilage is possible in conventional radiology and tomography and the decrease in joint space is only indirect evidence for joint destruction. CT is a radiologic method for direct visualization of joint cartilage but its diagnostic precision in the evaluation of early cartilage lesions is limited because it can not produce an image in a plane other than the transversal plane perpendicular to the direction of the main axis of the body and because it has limited spatial resolution. Other methods for direct visualization of joint cartilage are arthrography and arthroscopy which are little used in clinical practice because of their invasiveness and limited indications. MRI is a promising technique but its usage is limited by the high price and limited accessibility. Our object in the present study was to evaluate the significance of arthro-sonography in the diagnosis of early arthritic lesions of knee cartilage, based on the ultrasonographic assessment of the joint surface and cartilage thickness. Femoral cartilage was our choice for the study as there is statistically well documented high incidence of early arthritic changes in this area; moreover, the area is easily accessible for ultrasound evaluation using a scan perpendicular to the articular surface, incl. the cases with complete flexion of the knee joint, where the pressure areas of the condyles are apositioned to the tibial plateau. Using a 7.5 MHz transducer we managed to measure and document early arthritic changes in joint thickness and the contour of the joint surfaces before they can be detected using routine radiologic methods. PMID- 15359510 TI - Intraarticular rheumatoid nodule detection in the knee joint using ultrasonography. AB - Rheumatoid nodules represent a classic diagnostic sign of rheumatoid arthritis. They can be found in 20-25% of patients and are localized to the extensor surface of the proximal ulna and other friction areas such as the back of the head, the sacrum, the digits and the Achilles tendon. They can also develop in some internal organs such as the lungs, kidneys and the heart. Rheumatoid nodules are a characteristic extra-articular feature of RA. Intraarticular rheumatoid nodules are rare and their detection is a diagnostic challenge. There are few reports in the medical literature on their ultrasonographic detection. We report our results from a retrospective study on 9 patients (5 with gait problems and 4 - asymptomatic) with intraarticular rheumatoid nodules in the knee joint. Ultrasonography was most important in their detection. The diagnosis was confirmed histologically in two of our patients following surgical removal of the nodules. PMID- 15359511 TI - Immune profile and thyroid function in patients with rheumatoid arthritis. AB - Thyroid autoantibodies and thyroid function were studied in 175 patients with rheumatoid arthritis (ARA criteria, 1987). The study included 149 women and 26 men aged (54.7 +/- 13.1) all treated in the Clinic of Rheumatology, Higher Medical Institute, Plovdiv. A control group comprising 72 healthy subjects (aged 51.2 +/- 3.2) was used. Rheumatoid factor (RF) (latex agglutination test and Waaler-Rose test), antithyroglobulin (TAT) and antimicrosomal antibodies (MAT), thyroid stimulating hormone (TSH) and the free fraction of thyroxine (FT4) were measured. 69.7% of all patients tested positive for RF by the latex test and 64.5% by the Waaler-Rose test. TAT was found in 55 (31.4%), MAT in 26 (14.8%) from all the examined, which is significantly greater in comparison with the controls (p < 0.05). The thyroid gland was in a euthyroid state in most of the RA patients although MAT and/or TAT were detected in them. The results suggest existence of two autoimmune diseases - rheumatoid arthritis and Hashimoto's thyroiditis and raise the question of whether they are not manifestations of a single autoimmune disease with different body organ localisation. PMID- 15359512 TI - Differential diagnosis of Kikuchi's disease and systemic lupus erythematosus lymphadenopathy: clinicopathologic algorithm. AB - The authors have described the first case of Kikuchi's disease in Bulgaria. The present study gives the clinicopathologic algorithm compiled by them to make the correct diagnosis and differentiate Kikuchi's disease from systemic lupus erythematosus lymphadenopathy. PMID- 15359513 TI - Wrist tuberculosis in cladribine--induced remission of hairy cell leukosis. AB - A 45-year-old man with "hand-shoulder" syndrome developing eight-months after Cladribine-induced remission of an 11-year-old hairy cell leukosis is presented. Wrist bone biopsy was performed because of failure of the algodystrophy treatment and radiographic findings of progressive osteoporosis. Caseating epiteloid granulomas abundant in Langhans cells were found histologically and later Mycobacterium tuberculosis species was isolated in culture specimen. Fistulas were formed that healed after a prolonged anti-tuberculosis therapy. The role of cellular immunity deficiency in Cladribine-treated hairy cell leukosis that predisposes to mycobacterial infection is discussed. PMID- 15359514 TI - Let's stop "supersizing" our children. PMID- 15359515 TI - This time it's personal. PMID- 15359516 TI - TB in the ICU. PMID- 15359517 TI - Safety is job one--Tennessee Medicine looks at Tennessee's patient safety measures. PMID- 15359518 TI - The big business of bad babies. PMID- 15359519 TI - Constipation not always benign. PMID- 15359520 TI - Bilateral globus pallidus internus deep brain stimulation therapy for primary generalized dystonia. AB - Primary generalized dystonia (PGD) associated with the early-onset generalized dystonia gene (DYT1) can cause severe disability, compromising an individual's ability to perform activities of daily living. Pharmacological treatment has been inadequate in alleviating the motor dysfunctions. Deep brain stimulation of the bilateral globus pallidus internus (DBS B-GPi) has been documented to reduce these debilitating motor abnormalities. This report details the successful treatment of a DYT1-positive 13 year-old boy suffering from PGD. PMID- 15359521 TI - Epidemiology of bacterial enteric infections under active surveillance in Tennessee and Georgia, 2000-2002. PMID- 15359522 TI - Tort reform in medical malpractice. PMID- 15359523 TI - Improving mental health for ethnic older adults. PMID- 15359524 TI - Postintervention focus groups: toward sustaining care. AB - This study examined whether effects of person-centered care and person-centered mentoring were sustained postintervention from the perspective of long-term care nursing staff. Separate focus groups for administration, nurse mentors, and certified nursing assistants (CNAs) were used to assess perception of change in behaviors. CNAs sustained person-centered care skills on the job while nurse mentors experienced less change. With regard to nurse-CNA relationships, a discrepancy in the perceptions of nurses and CNAs was evident. The non threatening environment of focus groups allowed staff to share their perceptions of the barriers to behavior change needed to sustain the intervention over time. PMID- 15359525 TI - Elder care supportive interventions protocol: reducing discomfort in confused, hospitalized older adults. AB - This study tested the Elder Care Supportive Interventions Protocol (ECSIP), which are nursing and family support interventions designed to reduce discomfort and the associated consequences of delirium, impaired physical function, and need for post-hospital care in hospitalized older adults experiencing confusion from delirium or dementia. All adults older than age 74 or older than age 64 if from a nursing home who were admitted to the medical and surgical units of a tertiary care hospital were screened for a diagnosis of dementia, dependence on functional activities, or delirium. The ECSIP includes interventions to prevent and control discomfort, and to promote environmental organization and familiarity. Outcome measures administered within 48 to 72 hours of admission and again within 24 hours prior to discharge included the modified Discomfort Screen--Dementia Alzheimer's Type, NEECHAM Confusion Scale, and Katz Index of Activities of Daily Living. Patients problems with delirium and impaired physical function improved over hospitalization but were still pronounced at discharge. With the exception of reduced discomfort, there were no significant treatment effects for patients, which is attributed, in part, to inadequate implementation of the ECSIP protocol by nursing staff. PMID- 15359526 TI - Caring for family members with Alzheimer's disease: perspectives from Chinese American caregivers. AB - The purpose of this study was to examine the experiences of Chinese American caregivers who provide care for family members with Alzheimer's disease (AD). Individual interviews were conducted with four family caregivers. Results revealed ethnocultural and structural barriers that Chinese family caregivers experienced, including stigmatization of AD in the Chinese community, a lack of knowledge about AD, a lack of culturally and linguistically appropriate AD services, nega tive interactions with health-care providers, and difficulty with English. This study vides cultural knowledge and insights nurses who care for AD clients and their fam ilies. Overcoming barriers and working with Chinese American families and their community are needed to improve access and dementia services for Chinese American patients and their families. PMID- 15359527 TI - Treatment of substance misuse in older women: using a brief intervention model. AB - Alcohol and benzodiazepine misuse is a significant problem in older women for a number of reasons such as physiological changes, outdated prescribing practices, and failure to identify hazardous use. In addition, treatment barriers involving the health-care system, conflicting information, and ageism also exist. Substance misuse among older women is predicted to become a bigger problem as the baby boom generation ages. Brief interventions that consist of assessment, feedback, responsibility, advice, menu, empathy, and self-efficacy, or A-FRAMES, have the potential to reduce alcohol and benzodiazepine misuse among older women in a cost effective manner. PMID- 15359528 TI - Older married couples and health promotion: joint decision-making process for long-term spouses. AB - The purpose of this study was to explore communication patterns between long-term married couples to better understand the process by which later-life couples participate in health promotion. Forty community-residing couples married for 30 years or longer were asked to respond to open-ended questions and to participate in a decision-making activity. The majority of couples made health decisions jointly, with wives acting as the final deciders 54% of the time. Most couples used positive communication strategies during a structured scenario. Findings highlight the importance of expanding beyond traditional individual-focused models to include joint decision-making processes. Implications for future research and for nursing practice are discussed. PMID- 15359529 TI - Predictors of life satisfaction: perceptions of older community-dwelling adults. AB - The purpose of this study was to investigate variables that predict life satisfaction in elderly individuals. A convenience sample of 70 older community dwelling adults participated in the study. Instruments used included the Life Satisfaction Index A Scale, three subscales from the Self-Evaluation of Life Function Scale, the Perceived Control Scale, Hollingshead's Two Factor Index of Social Position, and the Self-Rated Health Subscale of the Philadelphia Geriatric Center Multilevel Assessment Instrument. When the influence of health and social position was held constant, social interaction was the only additional significant predictor of life satisfaction. When health and social position were not held constant, social interaction, perceived control, and social position were significant predictors of life satisfaction. The findings indicate that nursing practice, education, and research should focus on assessment and interventions relative to promoting and maintaining health, perceived control, and social interaction among elderly individuals living in the community. PMID- 15359530 TI - Geographic patterns of (genetic, morphologic, linguistic) variation: how barriers can be detected by using Monmonier's algorithm. AB - When sampling locations are known, the association between genetic and geographic distances can be tested by spatial autocorrelation or regression methods. These tests give some clues to the possible shape of the genetic landscape. Nevertheless, correlation analyses fail when attempting to identify where genetic barriers exist, namely, the areas where a given variable shows an abrupt rate of change. To this end, a computational geometry approach is more suitable because it provides the locations and the directions of barriers and because it can show where geographic patterns of two or more variables are similar. In this frame we have implemented Monmonier's (1973) maximum difference algorithm in a new software package to identify genetic barriers. To provide a more realistic representation of the barriers in a genetic landscape, we implemented in the software a significance test by means of bootstrap matrices analysis. As a result, the noise associated with genetic markers can be visualized on a geographic map and the areas where genetic barriers are more robust can be identified. Moreover, this multiple matrices approach can visualize the patterns of variation associated with different markers in the same overall picture. This improved Monmonier's method is highly reliable and can be applied to nongenetic data whenever sampling locations and a distance matrix between corresponding data are available. PMID- 15359531 TI - Inbreeding patterns in La Cabrera, Spain: dispensations, multiple consanguinity analysis, and isonymy. AB - Marital structure and inbreeding coefficients were analyzed in La Cabrera, an isolated mountain region in northwestern Spain. A total of 5,714 marriages were celebrated from 1880 to 1989 in the 37 parishes of the area. The total frequency of consanguineous marriages (up to the fourth degree) is 23.05%; multiple consanguineous marriages are remarkably common, reaching 5.43% of the total. The first cousin/second cousin ratio (referred to as kinship-type frequencies) is 0.43. The inbreeding values are the highest recorded in Spain and in Europe: alpha3 is 4.82 x 10(-3) for the whole period and alpha4 is 6.78 x 10(-3) for 1880 -1919. The temporal trend of inbreeding shows high values (alpha3 > 4.5 x 10(-3)) for a particularly long period (1900--1959) and a rapid decline from 1960 onward. This historical inbreeding trend is clearly related to changes in population size. The frequencies of multiple consanguineous marriages and the analysis of isonymy show that the inbreeding structure is related to geographic and demographic factors. Comparing the results at two hierarchical levels (La Cabrera as a whole and the 37 parishes individually), we conclude that the inbreeding values are affected by internal geographic subdivision of the population (Wahlund effect). Social and cultural factors, such as avoidance of or preference for consanguineous marriages, are less important but depend on the kinship type involved. PMID- 15359532 TI - Patterns of ethnic, linguistic, and geographic heterogeneity of palmar interdigital ridge counts in the Indian subcontinent. AB - Published data on palmar interdigital ridge counts (a-b, b-c, and c-d) among 57 populations from the Indian subcontinent were analyzed with reference to ethnic, socioeconomic, linguistic, and geographic affiliations of the studied populations. The spatial autocorrelation analysis suggests significant correlation between dermatoglyphic and geographic distances. The congruence with the ethnic semblance of the groups is also apparent in the data, and, in fact, the multiresponse permutation procedure did suggest highly significant within group homogeneity, confirming the biological validity of the social and ethnic criteria used in the analysis. The plots of populations on the first two principal components, accounting for 92% of the total variance, complement and support the results based on the other analyses, which show certain ethnic and geographic patterns. These findings can serve as baseline information for future studies on population variation in India, particularly studies based on molecular genetic markers, a trend that has already gained momentum. PMID- 15359533 TI - Genetic history of the population of Corsica (western Mediterranean) as inferred from autosomal STR analysis. AB - To genetically reconstruct the demographic history of the human population of Corsica (western Mediterranean), we analyzed the variability at eight autosomal STR loci (FES, VWA, CSF1PO, TH01, F13A1, TPOX, CD4, and D3S1358) in a sample of 179 native blood donors from 4 out of the 5 administrative districts. The main line of genetic discontinuity inferred from the spatial distribution of STR variability overlapped the linguistic and geographic boundaries. In the innermost areas (Corte district) several estimators had larger stochastic effects on allele frequencies. Genetic distance measures underlying different evolutionary models all pointed to a higher variability within Corsicans than within the rest of the Mediterranean reference populations. All Corsican subsamples showed the highest distance with a pooled sample from central Sardinia, thus making recent gene flow between the two neighboring islands unlikely. Hierarchical AMOVA and distance based multivariate genetic spaces stressed the closeness of Tuscan and Corsican frequency distributions, which could reflect peopling events with different time depths. Anyway, estimated separation times well support the linguistic hypothesis that Neolithic/Chalcolithic events have been far more important than Paleolithic or historical processes in the shaping of present Corsican variability. PMID- 15359534 TI - Frequencies of apolipoprotein A4 gene polymorphisms and association with serum lipid concentrations in two healthy Spanish populations. AB - Apolipoprotein A4 (apoA4) plays a role in intestinal lipid absorption. Several experimental interventions have shown that common variations at residues 347 (Thr --> Ser) and 360 (Gln --> His) on apoA4 are associated with differences in plasma lipid response to dietary fat; however, association studies between these variants and plasma lipid concentrations in populations reveal mixed results. We examined the effects of these polymorphisms in 758 randomly selected subjects (mean age 36.7+/-9.5 years) from 2 Spanish regions differing in latitude and fat intake: Aragon and Comunidad Valenciana. Subjects were matched one to one by sex and age. Frequencies for the less common alleles were similar in both regions: 0.096 (95% CI: 0.111-0.081) for codon 360 and 0.196 (95% CI: 0.216-0.176) for codon 347. In men and women there was no association between the codon 360 polymorphism and total cholesterol or triglyceride levels. However, subjects carrying the 360His allele had low-density lipoprotein (LDL) cholesterol concentrations statistically lower than homozygotes for the 360Gln allele, even after further adjustment for sex, age, region, body mass index, and APOE polymorphism (p = 0.043). The less common allele at codon 347 (the 347Ser allele) was associated with increased LDL-cholesterol concentrations with a clear gene dosage effect after multivariate adjustment (p = 0.029). Although these polymorphisms showed no heterogeneity by geographic region, the magnitude of the effect was higher in subjects from Aragon compared with the Comunidad Valenciana, suggesting a possible influence of the higher fat intake in Aragon. In the combined association analysis subjects with the 360Gln/347Ser pseudohaplotype had the highest LDL-cholesterol concentrations, supporting the antagonistic effect between the 360His and the 347Ser alleles on this trait. PMID- 15359535 TI - Apolipoprotein E polymorphism and its association with serum lipid levels in Brazilian children. AB - The influence of apolipoprotein E (APOE) genotypes on plasma lipid levels was determined in 414 Brazilian healthy children of mixed ethnicity, age 5 to 15 years (mean 8.9+/-2.9). The APOE*3 allele was the most frequent (77%), followed by APOE*4 (17%) and APOE*2 (6%). The pattern of lipid differences among genotypes was similar in both boys and girls. We did not detect an increase in cholesterol levels with the presence of the APOE*4 allele. Because a growing body of evidence suggests that the effect of *4 depends on its interaction with diet, the frequency of *4 might be more variable in children than in adults as well as among populations. Children carrying a *2 allele had lower total cholesterol (TC) and LDL-cholesterol levels (138.47+/-24.32 and 77.72+/-19.42, respectively) compared with *3/*3 children (158.33+/-25.28 and 97.05+/-21.82, respectively). Mean TC/HDLC ratio was also lower in children with the APOE*2 allele (3.27+/-0.66 versus 3.64+/-0.76). In this highly admixed population sample, the *2 anti atherogenic lipid pattern is already detected in children. PMID- 15359536 TI - Genetic variation in a compound short tandem repeat/Alu haplotype system at the SB19.3 locus: properties and interpretation. AB - The genetic variation at a compound nonrecombining haplotype system, consisting of the previously reported SB19.3 Alu insertion polymorphism and a newly identified adjacent short tandem repeat (STR), was studied in population samples from Portugal and Sao Tome (Gulf of Guinea, West Africa). Age estimates based on the linked microsatellite variation suggest that the Alu insertion occurred about 190,000 years ago. In accordance with the global patterns of distribution of human genetic variation, the highest haplotype diversity was found in the African sample. This excess in African diversity was due to both a substantial reduction in heterozygosity at the Alu polymorphism and a lower STR variability associated with the predominant Alu insertion allele in the Portuguese sample. The high level of interpopulation differentiation observed at the Alu locus (F(ST) = 0.43) was interpreted under alternative selective and demographic scenarios. The need for compatibility between patterns of variation at the STR and Alu loci could be used to restrict the range of selection coefficients in selection-driven genetic hitchhiking frameworks and to favor demographic scenarios dominated by larger pre expansion African population sizes. Taken together, the data show that the SB19.3 Alu-STR system is an informative marker that can be included in more extended batteries of compound haplotypes used in human evolutionary studies. PMID- 15359537 TI - Maternal cigarette smoking, metabolic enzyme polymorphism, and developmental events in the early stages of extrauterine life. AB - The recent observation that maternal ACP1 genotype has an interactive effect with smoking on intrauterine development prompted us to search for a possible interaction effect between smoking and ACP1 genotype on haptoglobin (Hp) development in the neonatal period. ACP1 is a highly polymorphic protein tyrosine phosphatase involved in signal transduction of several growth factor receptors. The enzyme is composed of two isoforms, F and S. We studied 299 infants born in the Department of Obstetrics of the University Hospital of Rome La Sapienza. We found that an interaction between ACP1 genotype and smoking has an effect on haptoglobin development: A significant delay of haptoglobin development in infants born to smoking mothers is observed only in infants with the ACP1 *B/*B genotype, which shows the highest concentration of the ACP1 F isoform. The results indicate that the ACP1 genotype modifies the deleterious effects of smoking on development not only during intrauterine life but also during the early stage of extrauterine life. PMID- 15359538 TI - Genetic variants of the paraoxonases (PON1 and PON2) in the Chilean population. AB - We estimated the frequencies of PON1 and PON2 variants (linked genes) in two hospital samples taken from the northern (San Jose Hospital, SJH) and eastern (Clinica Las Condes, CLC) parts of Santiago, Chile, using the polymerase chain reaction followed by restriction endonuclease digestion. The two hospital samples have different degrees of Amerindian admixture (SJH, 34.5%; CLC, 15.9%), which is reflected in the observed frequencies of the PON1 *B allele (SJH, 43.1%; CLC, 33.7%) and the PON2*S allele (SJH, 86.3%; CLC, 77.6%); both allele frequencies are significantly different between samples. The frequencies of the combined PON1 PON2 genotypes *A/*B-*C/*C, *A/*B-*S/*S, and *B/*B-*S/*S and of the haplotypes PON*A,C and PON*B,S were significantly different between the SJH and CLC groups. None of the genotype frequencies deviated significantly from those predicted by the Hardy-Weinberg equation. No linkage disequilibrium was found between the PON1 alleles and any of the PON2 alleles in either group (all p > 0.05). In our samples 38.52% (SJH) and 26.25% (CLC) of chromosomes must have the haplotype PON*B,S, presumed to be related to the risk of coronary artery disease. Twenty four of 193 (12.4%) SJH individuals and 7 of 122 (5.7%) CLC individuals were homozygotes for this haplotype. Finally, our data indicate ethnic-group-dependent genetic differences in the vulnerability to toxic organophosphorus. PMID- 15359539 TI - Apolipoprotein A1 gene polymorphisms at the -75 bp and +83/ 84 bp polymorphic sites in healthy Omanis compared with world populations. AB - The relative frequencies of the *A allele of the APOA1 gene at -75 bp (M1-) and the C or T +83/+ 84 bp allele (M2-) varied significantly between populations. We found the frequencies of M1- and M2- to be 0.22 and 0.067, respectively, in 150 healthy Omanis. These frequencies were compared to frequencies found in other world populations. PMID- 15359540 TI - Frequency of the 35delG mutation in the GJB2 gene in samples of European, Asian, and African Brazilians. AB - Mutations in the GJB2 gene are a major cause of congenital deafness. One specific mutation, the 35delG mutation, has accounted for most of the GJB2 mutations detected in European populations and is one of the most frequent disease mutations identified so far. We evaluated the frequency of the 35delG mutation in DNA samples from Brazilians of European, Asian, and African ancestry. All DNA samples were screened for the 35delG mutation using an allele-specific PCR. This study shows that the frequency of a common mutation (35delG) is significantly lower in non-European populations. PMID- 15359541 TI - The technology behind Colgate Simply White Toothpaste. AB - Colgate Simply White Toothpaste is a new advanced tooth whitening dentifrice that can be used every day. The synergy of abrasive stain removal with activated hydrogen peroxide delivers excellent performance in the removal of extrinsic and intrinsic tooth stain. Colgate Simply White Toothpaste provides other oral health benefits that have become the cost-of-entry into the toothpaste market: caries protection, tartar control, fresh breath, and a preferred flavor. PMID- 15359542 TI - A clinical investigation of the tooth whitening efficacy of a new hydrogen peroxide-containing dentifrice. AB - OBJECTIVE: The objective of this four-week, double-blind clinical study was to assess the tooth whitening efficacy of a new dentifrice delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride, and sodium tripolyphosphate in a high-cleaning silica base (Test Dentifrice), relative to that of a control dentifrice containing 0.243% sodium fluoride in a silica base (Control Dentifrice). METHODOLOGY: Following a baseline oral soft tissue examination and tooth shade assessment using the Vita Shade Guide, qualifying adult male and female subjects from the New Delhi, India area were randomized into either the Test or Control Dentifrice group. The two groups were balanced for baseline mean Vita Shade Guide rank scores. All subjects were provided with their assigned dentifrice and a soft bristled adult toothbrush for home use. Subjects were instructed to brush their teeth for two minutes twice daily (morning and evening), using only the dentifrice provided, and to refrain from using any other oral hygiene products for the entire four weeks of the study. There were no restrictions regarding diet or smoking habits during the course of the study. Oral soft tissue and tooth shade evaluations for each subject were repeated after two and four weeks of product use. RESULTS: Forty-four (44) subjects complied with the protocol and completed the entire study. At the two- and four-week examinations, the Test Dentifrice group had statistically significant mean shade rank score reductions of 4.04 and 5.17, respectively, compared to baseline (p < 0.05), while the Control Dentifrice had respective reductions of 0.41 and 0.53, which were also statistically significant. The mean shade rank reductions in the Test group compared to those in the Control group at two and four weeks were 3.63 and 4.64, respectively, and were statistically significant (p < 0.05). CONCLUSION: A new whitening dentifrice delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride and sodium tripolyphosphate has significant tooth whitening effectiveness, producing a mean shade rank score reduction from baseline of 5.17 after four weeks' use. PMID- 15359543 TI - Comparative tooth whitening and extrinsic tooth stain removal efficacy of two tooth whitening dentifrices: six-week clinical trial. AB - OBJECTIVE: The objective of this six-week, examiner-blind clinical study was to assess the tooth whitening and extrinsic tooth stain removal efficacy of a new dentifrice delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride, and sodium tripolyphosphate in a high-cleaning silica base (Test Dentifrice), relative to that of commercially available hexametaphosphate-containing whitening dentifrice (Positive Control Dentifrice). METHODOLOGY: Following a baseline oral soft tissue examination and scoring of extrinsic tooth stain and tooth shade, qualifying adult male and female subjects from the Harrisburg, Pennsylvania area were randomized into either the Test or Positive Control Dentifrice group. The two groups were balanced for gender, extrinsic tooth stain, and tooth shade scores. All subjects were provided their assigned dentifrice and a soft-bristled adult toothbrush for home use. Subjects were instructed to brush their teeth for two minutes twice daily (morning and evening) using only the dentifrice provided, and to refrain from using any other oral hygiene products for the entire six weeks of the study. There were no restrictions regarding diet or smoking habits during the course of the study. Oral soft tissue, extrinsic tooth stain, and tooth shade assessments for each subject were repeated after two and six weeks of product use. All statistical tests were two sided and employed a level of significance of alpha = 0.05. RESULTS: Fifty-six (56) subjects complied with the protocol and completed the entire study. Compared to baseline at both the two- and six-week examinations, the Test Dentifrice group had statistically significant reductions in extrinsic tooth stain area and intensity, and statistically significant mean shade rank reductions, with a six-week reduction of 4.81. In contrast, at six weeks, the Positive Control dentifrice had a statistically significant increase in tooth stain area, a non-significant increase in tooth stain intensity, and a statistically significant mean 1.40 shade rank score reduction. At both the two- and six-week examinations, the Test Dentifrice group had statistically significantly lighter tooth shades and statistically significantly lower levels of extrinsic tooth stain area and intensity than the Positive Control Dentifrice group. CONCLUSION: A new whitening dentifrice delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride, and sodium tripolyphosphate in a high-cleaning silica base has significant efficacy in tooth whitening and extrinsic stain removal, and provides a statistically significantly greater level of efficacy for both tooth whitening and extrinsic tooth stain removal than the commercially available Positive Control Dentifrice. PMID- 15359544 TI - Comparative tooth whitening and extrinsic tooth stain prevention efficacy of a new dentifrice and a commercially available tooth whitening dentifrice: six-week clinical trial. AB - OBJECTIVE: The objective of this six-week, examiner-blind clinical study was to assess the tooth whitening and extrinsic stain prevention efficacy of a new dentifrice delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride, and sodium tripolyphosphate in a high-cleaning silica base (Test Dentifrice) relative to that of a commercially available hexametaphosphate-containing whitening dentifrice (Positive Control Dentifrice). METHODOLOGY: Following a baseline oral soft tissue examination and scoring of extrinsic tooth stain and tooth shade, qualifying adult male and female subjects from the Mississauga, Ontario, Canada area were randomized into either the Test or Positive Control Dentifrice group. The two groups were balanced for age, tobacco use, extrinsic tooth stain, and tooth shade scores. All subjects received a dental prophylaxis, and were provided with a supply of their assigned product and a soft-bristled adult toothbrush for use at home. Subjects were instructed to brush their teeth for two minutes twice daily (morning and evening) using only the dentifrice provided, and to refrain from using any other oral hygiene products for the entire six weeks of the study. There were no restrictions regarding diet or smoking habits during the course of the study. Oral soft tissue, extrinsic tooth stain and tooth shade assessments for each subject were repeated after two and six weeks of product use. All statistical tests were two-sided and employed a level of significance of alpha = 0.05. RESULTS: Seventy-six (76) subjects complied with the protocol and completed the entire study. At both the two-week and six-week examinations, the Test Dentifrice prevented statistically significant (p < 0.05) more tooth stain as compared to the Positive Control Dentifrice. Also, at the two-week and six-week examinations, the Test Dentifrice provided statistically significant (p < 0.05) greater tooth whitening as compared to the Positive Control Dentifrice (2.64 and 3.77 shade rank reductions, respectively). CONCLUSION: A new whitening dentifrice, delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride, and sodium tripolyphosphate in a high-cleaning silica base provides a statistically significant greater level of efficacy for both tooth whitening and prevention of extrinsic tooth stain than the commercially available Positive Control Dentifrice. PMID- 15359545 TI - Electronic medical records: perils of outsourcing and the privacy act 1988 (CTH). PMID- 15359546 TI - Workplace violence in health care. PMID- 15359547 TI - Wrongful life claim: Harriton (by her tutor) v Stephens; Waller (by his tutor) v James; Waller (by his tutor) v Hoolahan. PMID- 15359548 TI - Electronic health records: international, structural and legal perspectives. AB - The EHR is a database record that incorporates a patient's health care details from conception to death and which can be distributed over a number of sites or aggregated at a particular source. This article describes the function and concept of the EHR by relating it to other medical information technologies, parallel changes in health care delivery, and a holistic health information model. The article compares the progress that Europe, Australia and the United States have made in the journey towards EHR implementation and concludes by highlighting some of the costs, barriers and consequences associated with the transition to a comprehensive EHR system. PMID- 15359549 TI - Trustworthy shared electronic health records: recordkeeping requirements and HealthConnect. AB - HealthConnect is a proposed national electronic health record system, centred on electronic health event summaries, that capture all health encounters of those patients and health care professionals who "opt in" to the system. This article reports on key findings of an analysis of HealthConnect's data principles, systems and business architecture, from a records continuum perspective, and from recordkeeping requirements of reliability and authenticity. It concludes that HealthConnect lacks critical recordkeeping functionality and that inadequate policy with regard to ownership, consent and privacy impacts on the business and systems architecture, and consequently its ability to deliver trustworthy records. PMID- 15359550 TI - Ethical issues in HealthConnect's shared electronic health record system. AB - Many countries are in the process of implementing systems of shared electronic health records. This article explores some of the ethical concerns raised by Australia's proposed HealthConnect system which aims to create electronic event summaries of health information. Three areas of ethical concern relating to confidentiality, consent and the involvement of the private sector are examined. It is argued that unless the HealthConnect system is firmly grounded in policy based on ethical considerations, patients may not want to "opt in" to it. PMID- 15359551 TI - HealthConnect and the duty of care: a dilemma for medical practitioners. AB - This article asks whether medical practitioners' duty of care to their patients will encompass participation in the HealthConnect shared electronic records initiative. Medico-legal aspects of the HeathConnect scheme relating to the nature of shared electronic health record summaries (SEHRS) are examined, focusing on their function as an element of patient care and their ultimate purpose. The analysis is based on the premise that an incomplete and hence inaccurate shared electronic health record summary is clinically and legally more perilous than no record at all. PMID- 15359552 TI - HealthConnect and privacy: a policy conundrum. AB - A shared electronic health record is intrinsically privacy-invasive because it creates a comprehensive record for information-sharing. The author explains the significance of information privacy and why it is that health information warrants special protection. She also provides an overview of the existing regulatory framework and an evaluation of suggested options and proposals for addressing privacy-related issues. Her analysis of suggested consent models suggests that they ultimately involve a trade-off between privacy and the broader benefits promised by HealthConnect and that obtaining the right balance is essential if HealthConnect is to achieve optimal health outcomes. PMID- 15359553 TI - Paths toward reclamation: therapeutic jurisprudence and the regulation of medical practitioners. AB - Much about what used to be termed "disciplinary" investigations and hearings is being revisited in the modern era. Therapeutic jurisprudence enables informed and sensitive awareness to potentially therapeutic and counter-therapeutic effects of both investigations and hearings conducted by medical regulatory authorities. This article analyses key aspects of authorities' processes from the perspective of notifiers/complainants and practitioners. Using developments at the Victorian Medical Practitioners Board as a base, it addresses issues of both investigative procedures and decision-making at formal and informal hearings, as well as the ramifications of re-hearings for the integrity of peer review informed regulation. It argues that where reclamation of practitioners is possible (namely where impropriety is not of the most serious order), there is much that is constructive about a focus upon enhancement of performance and competence levels, rather than the traditional preoccupation with whether registered status needs to be affected as a result of practitioner conduct. PMID- 15359554 TI - Stalking by law: damaging victims and rewarding offenders. AB - The introduction of anti-stalking statutes throughout Australia and much of the Western world has brought greater protection for the victims of various forms of harassment and intrusions. However, despite these new provisions, the legal system is failing some victims of stalking. This article examines those areas of the justice system that are particularly susceptible to manipulation by stalkers and the impact of these abuses on stalking victims. It also presents ways in which the problems encountered by stalking victims may be frustrated rather than alleviated by specific aspects of the enforcement of anti-stalking laws and of the functioning of certain courts and tribunals. Approaches that prevent or discourage the perpetuation of harassment and damage to victims of stalking within the legal system are discussed. PMID- 15359556 TI - Culling bad apples, blowing whistles and the Health Practitioners Competence Assurance Act 2003 (NZ). AB - The Health Practitioners Competence Assurance Act 2003 (NZ) became New Zealand law on 18 September 2003. This article looks at the background to the Act and reasons for resistance to it. It discusses the record so far on the medical profession's self-regulation of fitness-to-practise issues and the legal duties of practitioners who have reason to believe a colleague is putting patients at risk. It looks at some of the changes the Act brings, compares the experience of English and American medico-legal reforms, where applicable, and attempts to draw some conclusions on the chances for successful change. PMID- 15359555 TI - When silence threatens safety: lessons from the first Canberra Hospital neurosurgical inquiry. AB - Despite widespread institutional and professional support, the recommendations of the Bristol Royal Infirmary Inquiry may be insufficient to reduce patient risk from impaired senior medical practitioners. Using the First Inquiry into Neurosurgical Services at the Canberra Hospital as a case study, this article argues that the Bristol-type recommendations--which emphasise reformulation of clinical governance structures, including early reporting of "sentinel events" and compulsory clinical audits--will be ineffective without a reformed institutional ethos that encourages open transparency and respect for those committed to such processes. Such reformulation may need to commence in medical education and involve new strategies including the use of portable digital technology to facilitate self-assessment of performance and immediate reporting of adverse incidents. PMID- 15359557 TI - Is the immobility of advanced dementia a form of lorazepam-responsive catatonia? AB - Patients with end-stage dementia typically are very immobilized. Could this state actually be a form of lorazepam-responsive catatonia? Catatonia has been documented following cerebrovascular accidents, head injury, HIV encephalitis, brain tumors, and multiple sclerosis. Identified anatomical substrates include frontal lobes, parietal lobes, limbic system, diencephalon, and basal ganglia. Given that Alzheimer's disease, vascular dementia, Lewy body dementia, corticobasal degeneration, frontotemporal dementia, and Parkinsonian dementia often have degeneration in some of the same areas, dramatic awakenings might be possible by giving lorazepam challenges to locked-in dementia patients. If even a small percentage were lorazepam responders, the impact worldwide would be tremendous. Serious consideration should be given to undertaking large-scale clinical trials. PMID- 15359558 TI - Alleviation of multiple abnormalities by galantamine treatment in two patients with dementia with Lewy bodies. AB - Two patients diagnosed with dementia with Lewy bodies (DLB) presented with prominent behavioral disturbances. The first patient experienced well-formed visual hallucinations, movement disorders, and cognitive decline and was increasingly resistant to care. He was treated with galantamine 4 mg bid, which was increased to 8 mg bid after four weeks. Improvements in behavior, attention, and cognition made it possible for this patient to be transferred back to long term care after 32 days. The second patient had increasing confusion, fluctuating cognition, agitation, and psychosis. He was given donepezil initially, then switched to galantamine 4 mg bid. He was assessed as stable for discharge with improved behavior control after three weeks. Both patients also were receiving an atypical antipsychotic (quetiapine) and an antidepressant (venlafaxine and sertraline, respectively). PMID- 15359559 TI - Genetic aspects of Alzheimer's disease, Pick's disease, and other dementias. AB - Although genetic testing is available for some degenerative diseases, in most types of dementia, both genetic and environmental factors are involved. Overall, dementing diseases can be either sporadic or inherited, and in general, the earlier the onset, the more likely a disease is to be inherited. Before genetic testing is performed, the ethical issues, such as the effect the tests might have on asymptomatic children, should be considered. The ethical use of DNA samples in research is another genetic testing issue to be considered. PMID- 15359560 TI - Cost effects of a specialized care center for people with Alzheimer's disease. AB - A retrospective cohort control study of three populations, 65 years of age or older, at the same institution estimated the incremental cost of Alzheimer's disease (AD). The AD population of the ambulatory Alzheimer's Disease Center (ADC) (n = 640) was matched by age, gender, ethnicity, and address to one with AD from the general internal medicine practice (AD-GM) (n = 419) and to a control group without AD (n = 5331)from the same general medicine practice. Medicare costs of all care for all diagnoses were obtained for 1998 and 1999. Mean per person annual Medicare costs were $19,418 for ADC, $18, 753 for AD-GM, and $12,085 for the control group. Incremental cost for ADC population was $7,333 and $6,668 for AD-GM population compared with the control group. Incremental cost was $665 (9.1 percent) higher for ADC than AD-GM. Higher non-AD hospitalizations and length of stay (LOS) by AD populations were the main cost drivers. PMID- 15359561 TI - Discussions about cognitive impairment in first medical visits: older patients' perceptions and preferences. AB - Primary care physicians are the first medical contact for most patients with early-stage dementia. However, little is known about older patients' desire for discussions about cognitive problems or the frequency of discussions about cognitive status during primary care visits. To investigate this question, older patients and accompanying individuals were interviewed separately following the first visit with a primary care physician in an outpatient geriatric medical practice. Patients indicated that memory was discussed in 62 percent of visits. When memory was not discussed, almost one-third of patients stated that they would have wanted to discuss it. Physicians were more likely than patients to initiate discussions about memory. Patient factors, including age, gender, the presence of an accompanying individual, number of diagnoses, and the patient's statement that cognitive function was a main goal of the visit, were examined as predictors of the likelihood of discussion of memory during the first visit. Patients who identified discussing cognitive function as a goal of the visit were more likely to have a discussion about memory than those who did not. PMID- 15359562 TI - Evaluating the effect of therapeutic gardens. AB - Sometimes the visitors to therapeutic gardens do not necessarily experience the garden the way the designers intended. A postoccupancy evaluation (POE) can reveal discrepancies between the designer's intentions and use of the environment by the actual visitors. A POE is a user-oriented assessment that elicits the opinions of the visitors to a building, facility, or environment. This paper describes a POE of therapeutic gardens at a multilevel care facility for the aged. The population included patients at various stages of Alzheimer's disease progression. PMID- 15359563 TI - Dying dementia patients: too much suffering, too little palliation. AB - Suffering is traditionally viewed as a state encompassing psychological distress, spiritual concerns, and various aspects of physical pain. There is insufficient clinical evidence for suffering in dying dementia patients, which may lead to inappropriate evaluation and insufficient palliative treatment. Our objective was to evaluate the suffering of terminal dementia patients over time, from admission to a geriatric ward to the last day of life. The study included consecutive end stage dementia patients dying in a general geriatric department of a tertiary hospital. Patients were evaluated weekly by the Mini Suffering State Examination scale (MSSE). Seventy-one patients were studied. Mean survival of patients was 38.0 +/- 5.1 days. MSSE increased during hospital stays from 5.62 +/- 2.31 to 6.89 +/- 1.95 (p < 0. 001). According to the MSSE scale, 63.4 percent and 29.6 percent of patients died with a high and intermediate level of suffering, respectively. Only 7 percent of the patients died with a low level of suffering. In particular, patients were restless (p < 0. 001), had pressure sores (p = 0. 01), and were considered medically unstable (p < 0. 001). We concluded that, despite traditional medical and nursing care, a large proportion of dying dementia patients experienced an increasing amount of suffering as they approached death. New palliative treatment approaches should be developed for these patients. PMID- 15359564 TI - Stress in the caregivers of Alzheimer's patients: an experimental investigation in Italy. AB - In Italy, the family constitutes the major caregiving response to the needs of the elderly who are no longer self-sufficient. These caregivers are often wives or daughters who have chosen to keep the patient at home with them. On average, three-quarters of the caregiver's day is devoted to the patient, a proportion that tends to increase linearly as the disease progresses. The primary aim of our study was to describe a group of Italian caregivers of patients with a diagnosis of dementia to assess their levels of stress. We then correlated these caregivers with a number of sociodemographic variables and the patients' degree of cognitive impairment and independence in daily life activities. The study was conducted on a sample group of 236 caregivers of patients with a diagnosis of Alzheimer's disease (AD) according to the DSM-IV criteria. Each caregiver took part in a sociodemographic interview and filled in two questionnaires: the Caregiver Burden Inventory (CBI), to quantify the caregiving workload and the Brief Symptom Inventory (BSI), to assess the level of anxiety and depression. Patients were administered the Mini-Mental State Examination (MMSE) to evaluate their level of cognitive impairment and the Instrumental Activities of Daily Living (IADL) and Activities of Daily Living (ADL) tests to quantify their level of independence. The analysis of the results shows that the average Italian caregiver of an AD patient is a woman, approximately 60 years old. The majority of caregivers are spouses, followed by children. In general, these caregivers work at home, are housewives or retired, and are personally involved in caring for the patients. The greater the level of a patient's cognitive impairment and the less independent they are, the greater amount of care and supervision they require, leaving less free time for the caregiver; this leads to higher levels of anxiety related to caregiving. Toward the final stages of the disease, patient care tasks take up nearly all of the caregiver's free time, leaving them only about two hours per week for themselves or their friends. In conclusion, the management of dementia patients places a particular burden on the caregiver and involves several economic and social costs. The burden becomes heavier as the disease progresses, since the increase of cognitive disorders and the resulting reduction of independence in daily life, together with the onset of behavioral symptoms, heighten the caregiver's distress, anxiety, and depression. Establishing a network of community services able to alleviate the burden on families is therefore a priority. Parallels can be drawn between the stresses documented for American caregivers and their Italian counterparts. PMID- 15359565 TI - The dance of love. PMID- 15359567 TI - The role of metabolites in bioequivalence. AB - The role of metabolites in bioequivalence studies has been a contentious issue for many years. Many papers have published recommendations for the use of metabolite data based on anecdotal evidence from the results of bioequivalence studies. Such anecdotal evidence has validity, but the arguments lack weight because the "correct" answers are always unknown. A more promising area of exploration is recommendations based on simulated bioequivalence studies for which the "correct" answers are known, given the assumptions. A review of the literature, however, reveals scant evidence of attempts to apply to real data the pharmacokinetic principles on which the recommendations from simulated studies relied. We therefore applied those principles (based on estimates of intrinsic clearance after oral administration of the parent drug) to four bioequivalence studies from our archives, in which the parent drug and at least one metabolite were monitored. In each case, the outcome is discussed in the context of the complexity of the metabolic processes that impact on the parent drug and the metabolite(s) during the first passage from the intestinal lumen to the systemic circulation. Our observation is that no simple generalization can be made such that each drug/metabolite combination must be examined individually. Our recommendation, however, is that in the interests of safety, bioequivalence decision-making should be based on the parent drug whenever possible. PMID- 15359568 TI - A compartment model for the membrane-coated fiber technique used for determining the absorption parameters of chemicals into lipophilic membranes. AB - PURPOSE: The purpose of this work was to develop a compartment model for the membrane-coated fiber (MCF) technique for determining the absorption parameters of chemicals into lipophilic membranes. METHODS: A polymer membrane coated onto a section of inert fiber was used as a permeation membrane in the MCF technique. When MCFs were immersed into a donor solution, the compounds in the solution partitioned into the membrane. At a given permeation time, a fiber was removed from the solution and transferred into a gas chromatography injector for quantitative analysis. The permeation process of a given chemical from the donor phase into the membrane was described by a one-compartment model by assuming first-order kinetics. RESULTS: A mathematical model was obtained that describes the cumulative amount of a chemical permeated into the membrane as a function of the permeation time in an exponential equation. Two constants were introduced into the compartment model that were clearly defined by the physiochemical parameters of the system (a kinetic parameter and the equilibrium absorption amount) and were obtained by regression of the experimental data sampled over a limited time before equilibrium. The model adequately described the permeation kinetics of the MCF technique. All theoretical predictions were supported by the experimental results. The experimental data correlated well with the mathematical regression results. The partition coefficients, initial permeation rate, uptake, and elimination rate constants were calculated from the two constants. CONCLUSIONS: The compartment model can describe the absorption kinetics of the MCF technique. The regression method based on the model is a useful tool for the determination of the partition coefficients of lipophilic compounds when it takes too long for them to reach permeation equilibrium. The kinetic parameter and the initial permeation rate are unique parameters of the MCF technique that could be used in the development of quantitative structure-activity relationship models. PMID- 15359566 TI - Functional expression and localization of P-glycoprotein in the central nervous system: relevance to the pathogenesis and treatment of neurological disorders. AB - The expression of membrane drug transport systems in the central nervous system plays an important role in the brain disposition and efficacy of many pharmacological agents used in the treatment of neurological disorders such as neoplasia, epilepsy, and HIV-associated dementia. Of particular interest is P glycoprotein, a membrane-associated, energy-dependent, efflux transporter that confers the multidrug resistance phenotype to many cells by extruding a broad range of xenobiotics from the cell, resulting in poor clinical outcomes. In addition, the expression pattern of P-glycoprotein has recently been suggested to play a key role in the etiology and pathogenesis of certain diseases such as Alzheimer's and Parkinson's diseases. This review will focus on the cellular localization, molecular expression, and functional activity of P-glycoprotein in several compartments of the central nervous system and address its relevance in the pathogenesis and pharmacological treatment of neurological disorders. PMID- 15359569 TI - Use of biophysical characterization in preformulation development of a heavy chain fragment of botulinum serotype B: evaluation of suitable purification process conditions. AB - PURPOSE: The purpose of this study was to investigate the physicochemical and structural characteristics of recombinant botulinum serotype B (rBoNTB(Hc)) under various conditions and to use the information in evaluating suitable purification process conditions. METHODS: The solubility of rBoNTB(Hc) was evaluated at pH 4, 5, 6 7.5, 8, and 9. Secondary structure was evaluated using circular dichroism, and conformational stability was monitored using highsensitivity differential scanning calorimetry. Hydrophobic interaction chromatography, size exclusion chromatography-high performance liquid chromatography (SEC-HPLC), sodium dodecyl sulfate-poly acrylamide gel electrophoresis (SDS-PAGE), peptide mapping, and UV spectroscopy were used to monitor stability under the various conditions. RESULTS: The secondary structure of rBoNTB(Hc) consists predominantly of beta sheets. Solubility of rBoNTB(Hc) was lowest at its pI and highest at low and high pH. In the presence of NaCl, however, solubility decreased with increase in pH. Conformational and chemical stability are improved below pH 7.5. In the presence of 150 mM NaCl at high pH, conformational and chemical stability of rBoNTB(Hc) are further decreased. The study suggests that the purification process should minimize exposure of rBoNTB(Hc) to high pH and salt conditions. CONCLUSIONS: Optimal stability of rBoNTB(Hc) is achieved at low pH. The biophysical and analytical studies provide us with an understanding of rBoNTB(Hc) stability behavior in solution and assists in developing efficient purification conditions. PMID- 15359570 TI - Effects of the chemical structure and the surface properties of polymeric biomaterials on their biocompatibility. AB - Polymeric biomaterials have extensively been used in medicinal applications. However, factors that determine their biocompatibility are still not very clear. This article reviews various effects of the chemical structure and the surface properties of polymeric biomaterials on their biocompatibility, including protein adsorption, cell adhesion, cytotoxicity, blood compatibility, and tissue compatibility. Understanding these aspects of biocompatibility is important to the improvement of the biocompatibility of existing polymers and the design of new biocompatible polymers. PMID- 15359571 TI - Characterization and in vivo study of sustained-release formulation of human growth hormone using sodium hyaluronate. AB - PURPOSE: Aiming at once-a-week injection, a novel sustained release formulation of recombinant human growth hormone (SR-hGH) using sodium hyaluronate was developed for the treatment of children who have growth failure due to the lack of adequate secretion of endogenous growth hormone. METHODS: SR-hGH was produced in the form of solid microparticle using a Niro spray dryer and characterized by Malvern particle size analysis, scanning electron microscopy (SEM), size exclusion chromatography (SEC), reverse phase-high-performance chromatography (RP HPLC), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After in vitro release test, pharmacokinetic and pharmacodynamic studies were carried out in beagle dogs. SR-hGH was dispersed in medium-chain triglyceride (MCT) and administered at a dose of 1.0 mg hGH/kg subcutaneously. RESULTS: SR-hGH microparticles were successfully produced with a mean particle size of 5.6+/-1.0 microm. Physicochemical analysis with SEC, RP-HPLC, and SDS-PAGE showed that hGH extracted from SR-hGH was intact and comparable to that of hGH bulk standard indicating no structural change in hGH during the formulation processes. Monomeric content of hGH recovered from SR-hGH was 97.4% by SEC analysis, and its purity was 96% by RP-HPLC analysis. In vitro release test showed the sustained release characteristics of SR-hGH up to 48 h with the complete release of hGH loaded. The continuous and monotonous release profile observed in in vitro release test was supported by pharmacokinetic study in beagle dogs. Delayed absorption of hGH was observed with Cmax of 69.5+/-8.0 ng/ml and Tmax between 10 and 12 h. The administration of SR-hGH induced elevation of serum insulin-like growth factor-I (IGF-I) level for 6 days with a maximum value higher than the predose level by ca. 350 ng/ml. After 6 days, IGF-I level returned to the initial baseline level. CONCLUSIONS: Sustained-release formulation of hGH for once-a-week injection was successfully developed using high-molecular-weight sodium hyaluronate. No adverse effect was observed during and after the in vivo test using beagle dogs. PMID- 15359572 TI - In situ transport of vinblastine and selected P-glycoprotein substrates: implications for drug-drug interactions at the mouse blood-brain barrier. AB - PURPOSE: To study the intrinsic parameters of P-glycoprotein (P-gp) transport and drug-drug interactions at the blood-brain barrier (BBB), as few quantitative in vivo data are available. These parameters could be invaluable for comparing models and predicting the in vivo implications of in vitro studies. METHODS: The brains of P-gp-deficient mice mdr1a(-/-) and wild-type mice were perfused in situ using a wide range of colchicine, morphine, and vinblastine concentrations. The difference between the uptake by the wild-type and P-gp-deficient mice gave the P gp-linked apparent transport at the BBB. Drug-drug interactions were examined using vinblastine and compounds that bind to P-gp sites (verapamil, progesterone, PSC833) other than the vinblastine site to take into account the multispecific drug P-gp recognition. RESULTS: P-gp limited the brain uptake of morphine and colchicine in a concentration-independent way up to 2 mM. In contrast, vinblastine inhibited its own P-gp transport with an IC50 of approximately 56 microM and a Hill coefficient of approximately 4. The vinblastine efflux by P-gp was described by a Km at 16 microM and a maximal efflux velocity, Jmax, of approximately 8 pmol s(-1) g(-1) of brain. Similarly, vinblastine brain transport was increased by inhibiting P-gp as shown by the IC50 ranking, which was PSC833 < verapamil < vinblastine < progesterone. CONCLUSIONS: P-gp is responsible for both capacity-limited and -unlimited transport of P-gp substrates at the mouse BBB. In situ perfusion of mdr1a(-/-) and wild-type mouse brains could be used to predict drug-drug interactions for P-gp at the mouse BBB. PMID- 15359573 TI - Porcine ear skin as a model for the assessment of transdermal drug delivery to premature neonates. AB - PURPOSE: The purpose of this study was (i) to validate differentially tape stripped, porcine skin as an in vitro model for the evaluation of transdermal drug delivery (TDD) to premature neonates, (ii) to determine whether the model could estimate neonatal skin permeability as a function of postconceptional age (PCA), and (iii) to demonstrate that iontophoretic delivery permits precise control of drug input independent of skin barrier function. METHODS: Passive permeation of caffeine, phenobarbital, and lidocaine across tape-stripped porcine skin barriers was measured. Iontophoretic delivery of lidocaine across skins with different barrier competencies was also evaluated. RESULTS: For all drugs, passive permeation correlated with skin barrier function; that is, with transepidermal water loss (TEWL): Jss = A x exp[B x TEWL]. Combining this result with a previously derived dependence of TEWL upon the PCA of premature neonates in vivo allowed a relative value of Jss to be predicted for a given PCA. Comparison of these predictions showed excellent agreement with experimental data reported for diamorphine. Iontophoretic lidocaine delivery was precisely controllable independent of barrier competency. CONCLUSIONS: Porcine skin, in vitro, differentially tape-stripped to specific barrier competencies, is a useful model to explore TDD in premature neonates. The potential for iontophoresis to provide improved dose control and adjustment, irrespective of skin barrier maturity, is established. PMID- 15359574 TI - Transport characteristics of fexofenadine in the Caco-2 cell model. AB - PURPOSE: To investigate the membrane transport mechanisms of fexofenadine in the Caco-2 model. METHODS: Transport studies were performed in Caco-2 cell monolayers 21-25 days after seeding. The apparent permeability (Papp) of fexofenadine was determined in the concentration range 10-1000 microM in the basolateral-to-apical (b-a) and 50-1000 microM in the apical-to-basolateral (a-b) direction. The concentration-dependent effects of various inhibitors of P-glycoprotein (P-gp) (GF120918, ketoconazole, verapamil, erythromycin), multidrug resistant associated protein (MRP) (indomethacin, probenecid), and organic anion transporting polypeptide (OATP) (rifamycin SV) on the bidirectional transport of 150 microM fexofenadine were also examined. RESULTS: Fexofenadine displayed polarized transport, with the Pappb-a being 28- to 85-fold higher than the Papp(a-b). The Papp(a-b) was independent of the concentration applied, whereas Pappb-a decreased with increasing concentration (Vmax = 5.21 nmol cm(-2)s(-1) and K(M) = 150 microM), suggesting saturation of an apical efflux transporter. All four P-gp inhibitors had a strong, concentration-dependent effect on the Papp of fexofenadine in both directions, with GF 120918 being the most specific among them. The IC50 of verapamil was 8.44 microM on the P-gp-mediated secretion of fexofenadine. The inhibitors of OATP or MRP appeared not to affect the Papp(a-b) of fexofenadine in the Caco-2 model. CONCLUSIONS: This study clearly indicates that P-gp was the main transport protein of fexofenadine in the Caco-2 model. Even though P-gp was completely inhibited, fexofenadine was predicted to have a low fraction dose absorbed in humans due to poor intestinal permeability, and low passive diffusion seems to be the major absorption mechanism. PMID- 15359575 TI - Susceptibility to lipase-mediated digestion reduces the oral bioavailability of danazol after administration as a medium-chain lipid-based microemulsion formulation. AB - PURPOSE: To investigate the impact of lipidic formulation type on in vitro dispersion and digestion properties and the relationship to oral bioavailability, using danazol as a model lipophilic poorly water-soluble drug. METHODS: Three lipid-based danazol formulations [a long-chain triglyceride solution (LCT solution) and self-microemulsifying drug delivery systems (SMEDDS) based on long chain (C18) lipids (LC-SMEDDS) and medium-chain (C8-C10) lipids (MC-SMEDDS)] were administered to fasted beagle dogs and compared with a micronized danazol formulation administered postprandially and in the fasted state. In vitro dispersion and particle size data for the two SMEDDS were compared, and the distribution/solubilization patterns of danazol across the various phases produced during in vitro digestion quantified. RESULTS: The LCT-solution and LC SMEDDS formulations significantly enhanced the oral bioavailability of danazol when compared to fasted administration of the powder formulation. In contrast, and despite displaying excellent dispersion properties, the MC-SMEDDS resulted in little enhancement in danazol bioavailability. In support of the in vivo findings, in vitro digestion of the medium-chain formulation resulted in significant drug precipitation when compared with the long-chain lipid formulations. CONCLUSIONS: Digestion of microemulsion preconcentrate formulations based on medium-chain lipids may limit in vivo utility when compared with similar formulations based on long chain lipids. PMID- 15359576 TI - Influence of the type of surfactant and the degree of dispersion on the lymphatic transport of halofantrine in conscious rats. AB - PURPOSE: To compare the lymphatic transport and the portal absorption of halofantrine (Hf) when administered in (i) a triacylglycerol (TG) solution, (ii) an o/w-emulsion that contains a metabolizable surfactant, and (iii) an o/w emulsion that contains a synthetic surfactant. METHODS: Lymph cannulated rats were orally dosed with Hf in a TG solution or in o/w-emulsions dispersed by lecithin or Cremophor RH40. Lymph was continuously collected, and blood was sampled periodically in the course of 30 h. Hf in the lymph and blood and TG and phosphatidyl choline (PC) in the lymph were analyzed. RESULTS: A significantly (p < 0.05) higher level of Hf was found in the intestinal lymph when dosed in one of the emulsions (22.8+/-2.8% and 20.2+/-2.5%) compared to in the TG solution (7.9+/ 1.1%). No difference in the lymphatic transport of Hf was observed between the two emulsions. The portal absorption of Hf was similar for the three vehicles. CONCLUSIONS: The emulsified vehicles favor an increased lymphatic transport of Hf. The portal transport of Hf was not significantly different for the three vehicles. This indicates that a different degree of dispersion of the TG vehicle can change the route of transportation of Hf. PMID- 15359577 TI - Solvent exchange method: a novel microencapsulation technique using dual microdispensers. AB - PURPOSE: A new microencapsulation method called the "solvent exchange method" was developed using a dual microdispenser system. The objective of this research is to demonstrate the new method and understand how the microcapsule size is controlled by different instrumental parameters. METHOD: The solvent exchange method was carried out using a dual microdispenser system consisting of two ink jet nozzles. Reservoir-type microcapsules were generated by collision of microdrops of an aqueous and a polymer solution and subsequent formation of polymer films at the interface between the two solutions. The prepared microcapsules were characterized by microscopic methods. RESULTS: The ink-jet nozzles produced drops of different sizes with high accuracy according to orifice size of a nozzle, flow rate of the jetted solutions, and forcing frequency of the piezoelectric transducers. In an individual microcapsule, an aqueous core was surrounded by a thin polymer membrane; thus, the size of the collected microcapsules was equivalent to that of single drops. CONCLUSIONS: The solvent exchange method based on a dual microdispenser system produces reservoir-type microcapsules in a homogeneous and predictable manner. Given the unique geometry of the microcapsules and mildness of the encapsulation process, this method is expected to provide a useful alternative to existing techniques in protein microencapsulation. PMID- 15359578 TI - Physicochemical parameters associated with nanoparticle formation in the salting out, emulsification-diffusion, and nanoprecipitation methods. AB - PURPOSE: The aim of this work was to relate the physicochemical properties of the aqueous and organic phases used for nanoparticle (NP) preparation to the formation of NP produced by salting-out, emulsification-diffusion, and nanoprecipitation. METHODS: Methacrylic acid copolymer and poly(vinyl alcohol) (PVAL) were selected as NP polymer and emulsifying agent, respectively. Salting out and emulsification-diffusion NP batches were prepared modifying the PVAL content in the aqueous phase. For nanoprecipitation, NP were produced with variation of the polymer content and type of solvent in the organic phase. RESULTS: For salting-out and emulsification-diffusion, NP formation was discussed in terms of the emulsification theory. The nanoemulsion obtained during NP preparation was visualized by scanning electron microscopy. Aqueous and organic phases used for NP preparation were characterized by their viscosity and surface tension. NP characteristics such as particle mean size, residual surfactant, suspendability in water after freeze-drying, and morphology were explained in terms of these properties. For nanoprecipitation, NP formation was analyzed considering the diffusion-stranding phenomenon. CONCLUSIONS: NP formation by salting-out and emulsification-diffusion was related to PVAL chain interactions at the droplet interface (e.g., reduction in the interfacial tension, mechanical stabilization, and steric stabilization) and in the bulk solution (hydrodynamic stabilization). For nanoprecipitation, chi(solvent-water) and delta(delta solvent water) of the organic phase solvents were well related to the NP characteristics. PMID- 15359579 TI - Inverse gas chromatography: investigating whether the technique preferentially probes high energy sites for mixtures of crystalline and amorphous lactose. AB - PURPOSE: Inverse gas chromatography (IGC) is rapidly gaining popularity as a method for assessing powder surface energy. It is vital to understand what IGC measures if results are to be useful. This work examines the view that IGC preferentially measures high-energy sites on a powder surface. METHODS: Mixtures of amorphous (high-energy) and crystalline (lower energy) lactose particles were prepared and tested using IGC with nonpolar probes. The surface area of the particles was measured in situ in the inverse gas chromatograph. RESULTS: The results were weighted averages of the surface energy for amorphous and crystalline material until the amorphous content exceeded 15% w/w of the sample, after which the surface energy become equivalent to that of the amorphous form. The amorphous content dominated when the surface area was 40% of the total area. Given that the amorphous particles were much smaller and adhered to the crystalline ones, it is reasonable to conclude that many (most) of the binding sites on the surface of the crystalline particles were masked by the amorphous particles by the time that the amorphous content dominated the surface energy measurements. CONCLUSIONS: IGC does not simply measure the high-energy sites in the packed column, but equally there is a complex process that results in measured data on mixtures not being a weighted mean of the surface energy of the two components. PMID- 15359580 TI - Hydrophilic microspheres from water-in-oil emulsions by the water diffusion technique. AB - PURPOSE: In this study, we developed and evaluated a novel method to produce insulin-loaded hydrophilic microspheres allowing high encapsulation efficiency and the preservation of peptide stability during particle processing. The preparation method used the diffusion of water by an excess of solvent starting from a water-in-solvent emulsion. METHODS: The water dispersed phase containing albumin or lactose, or albumin-lactose in different weight ratios, and insulin was emulsified in water-saturated triacetin with and without emulsifiers, producing a water-in-triacetin emulsion. An excess of triacetin was added to the emulsion so that water could be extracted into the continuous phase, allowing the insulin-loaded microsphere precipitation. Insulin stability within the microspheres after processing was evaluated by reverse-phase and size-exclusion high-performance liquid chromatography. RESULTS: The water diffusion extraction process provided spherical microparticles of albumin or albumin-lactose. The mean diameter of the microspheres prepared with or without emulsifiers ranged from 2 to 10 microm, and the encapsulation efficiency of insulin was between 60% and 75%, respectively. The analysis of microsphere content after processing showed that insulin did not undergo any chemical modification within microspheres. The use of lactose alone led to the formation of highly viscous droplets that coalesced during the purification step. CONCLUSIONS: The water extraction procedures successfully produced insulin-loaded hydrophilic microspheres allowing the preservation of peptide stability. The type of excipient and the size of the disperse phase of the primary w/o emulsion were crucial determinants of microsphere characteristics. PMID- 15359581 TI - Identification of stabilized dynorphin derivatives for suppressing tolerance in morphine-dependent rats. AB - PURPOSE: Modulatory actions on morphine-induced effects, such as tolerance and withdrawal, have been noted for dynorphin A(1-13) [Dyn A(1-13)] and similar peptides. These are currently of limited therapeutic potential due to extensive metabolism by human metabolic enzymes resulting in a half-life of less than 1 min in human plasma. The purpose of this study was to identify stabilized dynorphin A (Dyn A) derivatives, to determine their metabolic routes in human plasma, and to assess whether the pharmacodynamic activity is retained. METHODS: The stability of peptides in human plasma was tested using in vitro metabolism studies with and without enzyme inhibitors. Identification of the generated metabolites was performed by mass spectrometry after high performance liquid chromatography (HPLC) separation. The in vivo activity of a stabilized dynorphin was tested by tail-flick assay in morphine-tolerant rats. RESULTS: Though amidation of the Dyn A(1-13) was able to stop the majority of C-terminal degradation, metabolism of Dyn A(1-10) amide continued by captopril sensitive enzymes, suggesting that Dyn A(1-13) amide is a better candidate for additional stabilization. Two Dyn A(1-13) amide derivatives further stabilized at the N-terminal end, [D-Tyr1]-Dyn A(1-13) amide and [N-Met-Tyr1]-Dyn A(1-13) amide, showed half-lives in plasma of 70 and 130 min, respectively. The most stable derivative [N-Met-Tyr1]-Dyn A(1-13) amide was tested successfully for retention of the pharmacological activity in modulating antinociceptive activity. CONCLUSIONS: [N-Met-Tyr1]-Dyn A(1-13) amide showed significant stability and antinociceptive activity in the tail-flick test, thus pointing to the clinical potential of this derivative in the management of pain as well as its potential activity in suppressing opiate tolerance and withdrawal. PMID- 15359582 TI - Evaluating barriers to bioavailability in vivo: validation of a technique for separately assessing gastrointestinal absorption and hepatic extraction. AB - PURPOSE: The purpose of this study was to develop and validate a method for separately evaluating the roles of gastrointestinal absorption and hepatic extraction as barriers to oral bioavailability (BA). The method was validated using five reference compounds known to have different absorption and hepatic extraction properties. Dose-dependence was also investigated for one reference compound. METHODS: Five reference compounds, amoxicillin, antipyrine, atenolol, propranolol, and testosterone, were administered as a cassette intravenouly (IV), via the hepatoportal vein (IPV), intraduodenally (ID), and intracolonically (IC) to male Sprague-Dawley rats. Blood samples were taken at nine time points, and the compounds were extracted from plasma using solid phase extraction. Plasma concentrations of each compound were determined using Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS). Pharmacokinetic parameters including bioavailability were calculated for each compound for each route of administration. RESULTS: Testosterone BA was less than 10% by ID, IC, and IPV routes, due to high hepatic extraction, consistent with its high systemic clearance (63 ml x min(-1) x kg(-1)) and short terminal plasma half-life (23 min). The IPV BA of amoxicillin was 95%+/-6% indicating the absence of hepatic extraction in the rat, but with an ID BA of approximately 39% suggesting incomplete GI absorption to be the main barrier to bioavailability. Absorption was poor from the colon, demonstrating site-dependence consistent with literature reports of site-dependent absorption. Low oral BA of propranolol was due in part to first-pass hepatic extraction (IPV BA of 36%). The IPV BA of propranolol was dose-dependent, most likely due to saturation of the P450 enzymes. Atenolol was incompletely bioavailable due to incomplete intestinal absorption, with no contribution of hepatic first-pass metabolism. Antipyrine was highly bioavailable by all routes. CONCLUSIONS: This in vivo rat model is demonstrated to be useful for identifying and quantifying the causes of incomplete bioavailabilty. It separately evaluates intestinal absorption, hepatic extraction, and site dependent absorption. Concentration-dependence of saturable processes can also be examined. PMID- 15359583 TI - Population dose-response model for tadalafil in the treatment of male erectile dysfunction. AB - PURPOSE: To determine the population dose-response relationship for tadalafil during on-demand (as-needed) administration for treatment of erectile dysfunction (ED). METHODS: A total of 212 male patients with mild, moderate, or severe ED participated in a multicenter, randomized, double-blind, placebo-controlled, parallel-group study. Patients were randomized to receive placebo or 2, 5, 10, or 25 mg tadalafil, taken on demand over an 8-week period. Efficacy was assessed on the basis of questions 2 and 3 of the Sexual Encounter Profile (SEP) and questions 3 and 4 of the International Index of Erectile Function (IIEF) questionaires. These scores were modeled using logistic regression. A fifth patient response, the IIEF EF (erectile function) domain score, was modeled as a continuous variable. RESULTS: The dose-response relationship for each efficacy variable was best described with an Emax model, in which maximum effect increased with ED severity at baseline. Response scores increased substantially between 10 and 25 mg tadalafil doses, and the dose-response parameter estimates suggested possibly higher responses at even higher doses. CONCLUSIONS: Population dose response modeling of all five oucome measures indicated that efficacy in all ED severity groups in the studied population generally increased across the 2 to 25 mg tadalafil dose range. Estimates of maximal improvement (Emax) in the IIEF EF domain score were 7.5, 11.4, and 16.3 points for patients with mild, moderate, and severe ED, respectively. Corresponding tadalafil doses to attain half-maximal improvement (ED50 estimates) were 4.7 mg, 7.1 mg, and 10.1 mg. PMID- 15359584 TI - Enhancement of transfection efficiency through rapid and noncovalent post PEGylation of poly(dimethylaminoethyl methacrylate)/DNA complexes. AB - PURPOSE: The aim of this work was to develop a new strategy to introduce poly(ethylene glycol) (PEG) into methacrylate-based polymer/ DNA complexes in order to produce hemocompatible particles able to transfect cells in the presence of serum. METHODS: Atom transfer radical polymerization was used to synthesize a well-defined poly(2-(dimethylamino)ethyl methacrylate) homopolymer (PDMAEMA) and a poly(2-(dimethylamino)ethyl methacrylate-b-poly(ethylene glycol) alpha-methyl ether, omega-methacrylate) palm-tree-like copolymer (P(DMAEMA-b-MAPEG)). The complexes obtained by self assembly of the pCMVbeta plasmid and the polymers were used to transfect Cos-7 cells. Their physical properties--particle size and zeta potential--were characterized respectively by dynamic light scattering and electrophoretic mobility measurements. Ex vivo hemocompatibility was also determined. RESULTS: The PDMAEMA/pCMVbeta complexes transfected Cos-7 cells exclusively in the absence of serum. Although the P(DMAEMA-bMAPEG) copolymer had no transfection activity per se, the addition of the latter to pre-formed PDMAEMA/DNA complexes significantly enhanced the activity and allowed transfection even in the presence of serum. The presence of palm-tree-like copolymers also improved the hemocompatibility properties of the complexes. No effect on platelet counts was observed for P(DMAEMA-b-MAPEG)/pCMVbeta complexes, whereas a decrease of platelets was clearly observed when blood cells were incubated with PDMAEMA/pCMVbeta complexes. CONCLUSIONS: Such a synergistic effect of noncovalent PEGylation of poly(amino methacrylate)/DNA complexes allows a new and versatile approach to tune up transfection efficiency. PMID- 15359585 TI - How well can an idiotope peptide mimic replace its parent idiotype in a synthetic peptide vaccine? AB - PURPOSE: To determine whether a vaccine consisting of an idiotope peptide mimic of the third complementarity-determining region of the immunoglobulin heavy chain (CDR-H3) is an effective substitute for its parent idiotype. Such peptide vaccines could ultimately be used for targeting pathological B lymphocytes. METHODS: Hen egg lysozyme (HEL) conjugates of the Fab' fragment of monoclonal anti-fluorescein antibody 9-40 (Fab'-HEL) or a peptide mimic of the 9-40 CDR-H3 (referred to as the "B epitope" or "Bep," the conjugate is referred to as "Bep HEL") were injected into separate cohorts of B10.A mice. Two additional control cohorts were injected with either the Bep peptide alone or a noncovalent mixture of Bep and HEL. Sera were assayed for both anti-idiotope and anti-idiotype activity by enzyme-linked immunosorbant assay (ELISA). Primary, secondary, and tertiary immune responses were examined. RESULTS: Both the Bep-HEL idiotope and the Fab-HEL idiotype immunogens elicited homologous, allogenic immune responses. No cross-reactivity was observed between anti-idiotope and anti-idiotype responses after primary immunization. With secondary immunization, 50% of mice immunized with the Bep-HEL conjugate exhibited a cross-reactive anti-idiotype response. Conversely, 100% of mice immunized with the Fab'-HEL conjugate exhibited a marginal, but statistically significant cross-reactive anti-idiotope response. Upon tertiary immunization, 100% of mice immunized with Bep-HEL exhibited a cross-reactive anti-idiotype response, and 55.6% of mice immunized with the Fab'-HEL conjugate exhibited a cross-reactive anti-idiotope response. CONCLUSIONS: Covalent coupling of a xenogenic carrier protein to an idiotype immunogen or its peptide mimic significantly enhances the intensity of homologous, allogenic anti-idiotype or anti-idiotope immune responses. Multiple immunizations are necessary to induce cross-reactivity between the peptide mimic and its parent idiotype. PMID- 15359586 TI - Evidence for modulation of P-glycoprotein-mediated efflux by methoxypolyethylene glycol-block-Polycaprolactone amphiphilic diblock copolymers. AB - PURPOSE: The purpose of this study was to investigate the pathways involved in rhodamine 123 (R-123) accumulation enhancement in Caco-2 cells with a low molecular-weight methoxypolyethylene glycol-block-polycaprolactone (MePEG-b-PCL) diblock copolymer. METHODS: R-123 accumulation by Caco-2 cells with MePEG17-b PCL5 was measured in the presence of endocytosis inhibitors or under ATP depletion conditions. Directional flux studies were conducted with cell monolayers on Transwell plates. RESULTS: Endocytosis inhibitors had no effect on reducing R-123 accumulation with MePEG17-b-PCL5. The apical to basolateral (AP- >BL) flux of R-123 with MePEG17-b-PCL5 or verapamil was similar to R-123 alone. However the BL-->AP flux was significantly decreased with MePEG17-b-PCL5 and verapamil. The efflux ratio for R-123 flux was 3.2 and was reduced to 1.06 with MePEG17-b-PCL5 confirming the inhibition of P-glycoprotein (P-gp)-mediated efflux. R-123 accumulation at the conclusion of each of the flux experiments was similar for MePEG17-b-PCL5 and verapamil in the BL-->AP direction. The AP-->BL direction demonstrated a 2-fold increase for verapamil and a 5-fold increase with MePEG17-b-PCL5. This difference in R-123 accumulation was possibly due to the diblock enhancing passive membrane diffusion of R-123. CONCLUSIONS: MePEG17-b PCL5 diblock reduced R-123 efflux through inhibition of P-gp efflux, and unimers may interact with cell membranes, increasing permeability and enhancing R-123 influx through increased transmembrane diffusion. PMID- 15359587 TI - The mechanism of protraction of insulin detemir, a long-acting, acylated analog of human insulin. AB - PURPOSE: Insulin detemir has been found in clinical trials to be absorbed with very low variability. A series of experiments were performed to elucidate the underlying mechanisms. METHODS: The disappearance from an injected subcutaneous depot and elimination studies in plasma were carried out in pigs. Size-exclusion chromatography was used to assess the self-association and albumin binding states of insulin detemir and analogs. RESULTS: Disappearance T50% from the injection depot was 10.2+/-1.2 h for insulin detemir and 2.0+/-0.1 h for a monomeric acylated insulin analog. Self-association of acylated insulin analogs with same albumin affinity in saline correlated with disappearance rate and addition of albumin to saline showed a combination of insulin detemir self association and albumin binding. Intravenous kinetic studies showed that the clearance and volume of distribution decreased with increasing albumin binding affinity of different acylated insulin analogs. CONCLUSIONS: The protracted action of detemir is primarily achieved through slow absorption into blood. Dihexamerization and albumin binding of hexameric and dimeric detemir prolongs residence time at the injection depot. Some further retention of detemir occurs in the circulation where albumin binding causes buffering of insulin concentration. Insulin detemir provides a novel principle of protraction, enabling increased predictability of basal insulin. PMID- 15359589 TI - Case managers likely to play big role in pay-for-performance initiatives. Hospital pay-for-performance measures being tested by CMS. PMID- 15359588 TI - Insertion and partition of sodium taurocholate into egg phosphatidylcholine vesicles. AB - PURPOSE: To get a continuous description of the insertion and partition processes of sodium taurocholate (TC) into the lipid bilayers of vesicles that can serve as a model for understanding the mechanism of destabilization by the bile salts of liposomes used as drug carriers for oral administration. METHODS: The progressive solubilization of egg phosphatidylcholine vesicles during TC addition at controlled rates was followed by continuous turbidity (OD) and resonance energy transfer (RET) between two fluorescent probes. The influence of the lipid and TC concentrations as well as the rate of TC addition on the processes were examined. RESULTS: Continuous turbidity recordings allowed following of the size and composition evolutions of the mixed TC/lipid aggregates formed at different steps of the vesicle-micelle transition. The solubilization mechanism is governed by complex kinetics that depend on the surfactant concentration and its addition rate. A two-step process characterizes the evolution of the vesicular state: interaction of TC molecules with the external monolayer of the vesicles first occurs. The homogeneous distribution of TC within the lipid matrix after its insertion is a very slow process. A micellar structural reorganization is observed when TC is added rapidly. CONCLUSIONS: This work provides detailed information on the slow insertion and diffusion kinetics of TC in liposomal bilayers by using a dynamic study which mimics physiological phenomena of digestion. PMID- 15359590 TI - Case managers can lead the way in P4P initiatives. Start by identifying your top DRGs. PMID- 15359592 TI - Telephonic program cuts CHF readmissions in half. Nurses follow up after discharge. PMID- 15359591 TI - Hospitals improve CMS project performance. Case managers lead the multidisciplinary team. PMID- 15359593 TI - Compliance with protocols may improve outcomes. First large-scale safety study of CABG procedures. PMID- 15359594 TI - Clinical guidelines for palliative care published. Initiative recognizes growth, maturity of field. PMID- 15359595 TI - Do you keep stroke patients waiting too long? PMID- 15359596 TI - Three strategies to reduce overcrowding and gridlock. PMID- 15359597 TI - If nurses hoard patients, can you improve flow? PMID- 15359598 TI - Expression of the stilbene synthase (StSy) gene from grapevine in transgenic white poplar results in high accumulation of the antioxidant resveratrol glucosides. AB - When present, stilbene synthase leads to the production of resveratrol compounds, which are major components of the phytoalexin response against fungal pathogens of the plant and are highly bioactive substances of pharmaceutical interest. White poplar (Populus alba L.) was transformed with a construct containing a cDNA insert encoding stilbene synthase from grapevine (Vitis vinifera L.), under the control of the cauliflower mosaic virus (CaMV) 35S promoter, and a chimeric kanamycin resistance gene. Southern blot hybridization analysis demonstrated the presence and integration of exogenous DNA sequences in the poplar genome. Expression of the stilbene synthase-encoding gene in different transgenic lines was confirmed by Western blot and Northern analyses. Compared to the controls, in the transgenic plants two new compounds were detected and were identified as the trans- and cis-isomers of resveratrol-3-glucoside (piceid) by high-pressure liquid chromatography (HPLC), UV spectrophotometry, electrospray mass spectrometry (HPLC-ESI-MS) and enzymatic hydrolysis. Since poplar is a good biomass producer and piceids are accumulated in substantial amounts (up to 615.2 microg/g leaf fresh weight), the transgenic plants represent a potential alternative source for the production of these compounds with high pharmacological value. Despite the presence of piceid, in our experimental conditions no increased resistance against the pathogen Melampsora pulcherrima, which causes rust disease, was observed when in vitro bioassays were performed. PMID- 15359599 TI - Viable transgenic goats derived from skin cells. AB - The current study was undertaken to evaluate the possibility of expanding transgenic goat herds by means of somatic cell nuclear transfer (NT) using transgenic goat cells as nucleus donors. Skin cells from adult, transgenic goats were first synchronized at quiescent stage (G0) by serum starvation and then induced to exit G0 and proceed into G1. Oocytes collected from superovulated donors were enucleated, karyoplast-cytoplast couplets were constructed, and then fused and activated simultaneously by a single electrical pulse. Fused couplets were either co-cultured with oviductal cells in TCM-199 medium (in vitro culture) or transferred to intermediate recipient goat oviducts (in vivo culture) until final transfer. The resulting morulae and blastocysts were transferred to the final recipients. Pregnancies were confirmed by ultrasonography 25-30 days after embryo transfer. In vitro cultured NT embryos developed to morulae and blastocyst stages but did not produce any pregnancies while 30% (6/20) of the in vivo derived morulae and blastocysts produced pregnancies. Two of these pregnancies were resorbed early in gestation. Of the four recipients that maintained pregnancies to term, two delivered dead fetuses 2-3 days after their due dates, and two recipients gave birth to healthy kids at term. Fluorescence in situ hybridization (FISH) analysis confirmed that both kids were transgenic and had integration sites consistent with those observed in the adult cell line. PMID- 15359600 TI - Superoxide dismutase transgenes in sugarbeets confer resistance to oxidative agents and the fungus C. beticola. AB - Sugarbeets carrying superoxide dismutase transgenes were developed in order to investigate the possibility of enhancing their resistance to oxidative stress. Binary T-DNA vectors carrying the chloroplastic and cytosolic superoxide dismutase genes from tomato, were used for Agrobacterium-mediated transformation of sugarbeet petioles. The transgenic plants were subjected to treatments known to cause oxidative stress, such as the herbicide methyl viologen and a natural photosensitizer toxin produced by the fungus Cercospora beticola, namely cercosporin. The transgenic plants exhibited increased tolerance to methyl viologen, to pure cercosporin, as well as to leaf infection with the fungus C. beticola. PMID- 15359601 TI - Complementation of alpha-thalassaemia in alpha-globin knockout mice with a 191 kb transgene containing the human alpha-globin locus. AB - alpha-thalassaemia is an inherited blood disorder caused by a decrease in the synthesis of alpha-globin due to mutations in one or both of the alpha-globin genes located on human chromosome 16. A 191 kb transgene derived from a sequenced bacterial artificial chromosome (BAC) clone carrying the human alpha-globin gene cluster, together with about 100 kb of sequence upstream of DNase1 hypersensitive site HS-40 and 30 kb downstream of the alpha1-globin gene, was introduced into fertilised mouse oocytes by pronuclear microinjection. Three transgenic founder mice were obtained. Analysis of one transmitting line by fluorescent in situ hybridisation and quantitative PCR demonstrated a single copy integration of the human alpha-globin transgene on chromosome 1. Analysis of haemoglobins from the peripheral blood by cellulose acetate electrophoresis and high performance liquid chromatography (HPLC) demonstrated synthesis of human alpha-globin to about 36% of the level of each mouse alpha-globin locus. Breeding of transgenic mice with mice heterozygous for a knockout (KO) deletion of both murine alpha-globin genes showed that the human alpha-globin locus restored haemoglobin levels and red cell distribution width to normal in double heterozygous mice and significantly normalised other haematological parameters. Interestingly the human transgene also induced a significant increase in red cell production and haematocrit above wild type values. This is the first report demonstrating complementation of a murine alpha-globin KO mutation by human alpha-globin gene expression from an intact human alpha-globin locus. The transgenic mouse model described in this report should be very useful for the study of human alpha-globin gene regulation and for the development of strategies to down regulate alpha-globin production as a means of ameliorating the severity of beta-thalassaemia. PMID- 15359602 TI - Recombinant protein expression plasmids optimized for industrial E. coli fermentation and plant systems produce biologically active human insulin-like growth factor-1 in transgenic rice and tobacco plants. AB - Human insulin-like growth factor-1 (hIGF-1) is a growth factor with clinical significance in medicine. The therapeutic potential of recombinant hIGF-1 (rthIGF 1) stems from the fact that hIGF-1 resembles insulin in many aspects of physiology. The expression of hIGF-1 in transgenic tobacco and rice plants using different expression cassettes is reported here. In the present study, two coding sequences were tested, one with the original human sequence, but partially optimized for expression in E. coli and the other with a plant-codon-optimized sequence that was expected to give a higher level of expression in plant systems. Three different hIGF-1 recombinant expression constructs were generated. All expression constructs utilized the maize ubiquitin 1 promoter with or without a signal sequence. Analyses conducted using a hIGF-1 specific ELISA kit showed all transgenic plants produced hIGF-1 and the accumulated hIGF-1 increased from the E. coli codon bias to higher levels when the hIGF-1 coding sequence was codon optimized to match that of the maize zeamatin protein--the most transcribed gene in maize endosperm suspension cells. Further analyses that compared the functionality of the bacterial signal peptide Lam B in plants showed that this leader peptide led to lower expression levels when compared to transgenic plants that did not contain this sequence. This indicated that this expression construct was functional without removal of the bacterial signal sequence. The maize ubiquitin 1 promoter was found to be more active in rice plants than tobacco plants indicating that in this case, there was a class preference that was biased towards a monocot host. Biological analyses conducted using protein extracts from transgenic plants showed that the rthIGF-1 was effective in stimulating the in vitro growth and proliferation of human SH-SY5Y neuroblastoma cells. This indicated that the plant-produced rthIGF-1 was stable and biologically active. As some plants have been reported to express an endogenous insulin-like protein, we also looked for any effect of the human growth factor in transgenic plants, but no developmental or morphological differences with wild type tobacco or rice plants were detected. Since insulin and hIGF-1 share some overlapping roles, hIGF 1 may become a substitute therapeutic agent in subjects with certain defects in their insulin receptor signaling. Hence, if the full beneficial potential of rthIGF-1 is achieved, it is expected that in the future the demand will likely increase significantly. PMID- 15359603 TI - Macrophage-specific overexpression of human matrix metalloproteinase-12 in transgenic rabbits. AB - Increased matrix metalloproteinase-12 (MMP-12) has been implicated in atherosclerosis and many other inflammatory processes. To define MMP-12 functions in vivo, we generated transgenic rabbits that expressed human (h) MMP-12 gene under the control of a macrophage-specific promoter, the human scavenger receptor promoter. Two transgenic founder rabbits were found to have hMMP-12 transgene integration by Southern blot analysis. hMMP-12 mRNA was expressed in peritoneal and alveolar macrophages, and in tissues enriched in macrophages in transgenic rabbits. High levels of hMMP-12 protein were detected in the conditioned media of cultured peritoneal and alveolar macrophages from transgenic rabbits. Zymography showed that hMMP-12 secreted from macrophages possessed enzymatic activity toward beta-casein. To evaluate the expression of hMMP-12 in inflammatory sites, we used carrageenan-induced granulomas as an in vivo model for tissue macrophages and foam cells. Granuloma size in transgenic rabbits was significantly increased compared to that in control rabbits, and histological examination revealed that granulomas of transgenic rabbits were enriched in macrophages associated with increased hMMP-12 expression. We believe that this transgenic rabbit model with increased expression of hMMP-12 may become a useful model for further mechanistic studies of MMP-12 in inflammatory diseases and cancer invasion; it is also an ideal model for testing the in vivo action of MMP-12 inhibitors. PMID- 15359604 TI - A novel two T-DNA binary vector allows efficient generation of marker-free transgenic plants in three elite cultivars of rice (Oryza sativa L.). AB - A pilot binary vector was constructed to assess the potential of the 2 T-DNA system for generating selectable marker-free progeny plants in three elite rice cultivars (ZhongZuo321, Ariete and Khao Dawk Mali 105) known to exhibit contrasting amenabilities to transformation. The first T-DNA of the vector, delimited by Agrobacterium tumefaciens borders, contains the hygromycin phosphotransferase (hpt) selectable gene and the green fluorescent protein (gfp) reporter gene while the second T-DNA, delimited by Agrobacterium rhizogenes borders, bears the phosphinothricin acetyl transferase (bar) gene, featuring the gene of interest. 82-90% of the hygromycin-resistant primary transformants exhibited tolerance to ammonium glufosinate mediated by the bar gene suggesting very high co-transformation frequency in the three cultivars. All of the regenerated plants were analyzed by Southern blot which confirmed co-integration of the T-DNAs at frequencies consistent with those of co-expression and allowed determination of copy number for each gene as well as detection of two different vector backbone fragments extending between the two T-DNAs. Hygromycin susceptible, ammonium glufosinate tolerant phenotypes represented 14.4, 17.4 and 14.3% of the plants in T1 progenies of ZZ321, Ariete and KDML105 primary transformants, respectively. We developed a statistical model for deducing from the observed copy number of each T-DNA in T0 plants and phenotypic segregations in T1 progenies the most likely constitution and linkage of the T-DNA integration locus. Statistical analysis identified in 40 out of 42 lines a most likely linkage configuration theoretically allowing genetic separation of the two T-DNA types and out segregation of the T-DNA bearing the bar gene. Overall, though improvements of the technology would be beneficial, the 2 T-DNA system appeared to be a useful approach to generate selectable marker-free rice plants with a consistent frequency among cultivars. PMID- 15359605 TI - Primary spermatocyte-specific Cre recombinase activity in transgenic mice. AB - We have evaluated the specificity of Cre recombinase activity in transgenic mice expressing Cre under the control of the synatonemal complex protein 1 (Sycp1) gene promoter. Sycp1Cre mice were crossed with the ROSA26 reporter line R26R, to monitor the male germ cell stage-specificity of Cre activity as well as to verify that Cre was not active previously during development of other tissues. X-gal staining detected Cre-mediated recombination only in testis. Detailed histological examination indicated that weak Cre-mediated recombination occurred as early as in zygotene spermatocytes at stage XI of the cycle of the seminiferous epithelium. Robust expression of X-gal was detected in early to mid late spermatocytes at stages V-VIII. We conclude that this transgenic line is a powerful tool for deleting genes of interest specifically during male meiosis. PMID- 15359607 TI - [Involvement of a public pharmacology laboratory in pharmacokinetic studies]. AB - Pharmacokinetic studies can be performed before or after a drug is marketed. Indeed, extensive use of a new drug can uncover situations that it has not been possible to investigate before market application. Hospital departments of pharmacology are often involved in these complementary studies. They can design the study, initiate it in a clinical investigation centre, measure plasma concentrations, analyse the pharmacokinetic data, and write the report. If the extent of the scientific involvement of public pharmacology laboratories is strictly demarcated, financial support for nonsponsored studies by the pharmaceutical industry is problematical. Indeed, it is difficult to obtain funds to study the concentration-toxicity relationship or a drug-drug interaction for a compound for which the patent has expired. In addition, such studies have an excess cost, mainly because of the expense associated with drug measurements. Also, the medical time devoted to research is not rewarded by institutions. This situation will become more complicated because of the restrictive legislative framework imposed by the European directive. PMID- 15359606 TI - Fimbrial subunit protein FaeG expressed in transgenic tobacco inhibits the binding of F4ac enterotoxigenic Escherichia coli to porcine enterocytes. AB - Plants offer a promising alternative for the production of foreign proteins for pharmaceutical purposes in tissues that are consumed as food and/or feed. Our long-term strategy is to develop edible vaccines against piglet diarrhoea caused by enterotoxigenic Escherichia coli (F4 ETEC) in feed plants. In this work, we isolated a gene, faeG, encoding for a major F4ac fimbrial subunit protein. Our goal was to test whether the FaeG protein, when isolated from its fimbrial background and produced in a plant cell, would retain the key properties of an oral vaccine, that is, stability in gastrointestinal conditions, binding to intestinal receptors and inhibition of the F4 ETEC attachment. For this purpose, tobacco was first transformed with a faeG construct that included a transit peptide encoding sequence to target the FaeG protein to the chloroplast. The best transgenic lines produced FaeG protein in amounts of 1% total soluble protein. The stability of the plant-produced FaeG was tested in fluids simulating piglet gastric (SGF) and intestinal (SIF) conditions. Plant-produced FaeG proved to be stable up to 2 h under these conditions. The binding and inhibition properties were tested with isolated piglet villi. These results showed that the plant produced FaeG could bind to the receptors on the villi and subsequently inhibit F4 ETEC binding in a dose-dependent manner. Thus, the first two prerequisites for the development of an oral vaccine have been met. PMID- 15359608 TI - Pharmacokinetics-pharmacodynamics during drug development--an example from Servier: ivabradine. AB - A short introduction to the principles of pharmacokinetic-pharmacodynamic (PK-PD) modelling and population approaches is provided in this article. The importance of implementing these techniques in the drug development process is illustrated by an example from experience at the Servier International Research Institute. This example demonstrates how the use of PK-PD modelling can rationalise the development process and save valuable time. Population approaches significantly contribute to the integration of PK-PD modelling into the different drug development phases by expanding the possibilities of application. PMID- 15359609 TI - [Biomarkers and pharmacokinetic-pharmacodynamic studies of immunosuppressive agents]. AB - Biomarkers are characteristics that are objectively measured and evaluated as indicators of normal biological processes, pathogenic processes, or pharmacological responses to a therapeutic intervention. Biomarkers of pharmacological responses allow pharmacokinetic-pharmacodynamic modelling based on what is known of the mechanism of action of the drug. Characteristics predictive of the pharmacological response, such as a genotype, constitute a specific category of biomarkers and can be used as covariates in the model. In the case of "classical" immunosuppressive agents, the biomarkers of response most studied are lymphocyte calcineurin activity for ciclosporin and tacrolimus, and inosine monophosphate dehydrogenase activity for mycophenolate. Pharmacokinetic pharmacodynamic analysis of monoclonal antibodies with immunosuppressive properties requires complex models and immunological biomarkers such as, for example, the number of circulating lymphocytes with a given surface antigen. Pharmacodynamic indirect response models are particularly relevant in this instance. PMID- 15359610 TI - Apoptosis: a target for neuroprotection. AB - Accumulating evidence strongly suggests that apoptosis contributes to neuronal death in a variety of neurodegenerative contexts. Activation of the cysteine protease caspase 3 appears to be a key event in the execution of apoptosis in the central nervous system. As a result, mice null for caspase 3 display considerable neuronal expansion, usually resulting in death by the second week of life. Consistent with the proposal that apoptosis plays a central role in human neurodegenerative disease, caspase-3 activation has recently been observed in stroke, spinal cord trauma, head injury and Alzheimer's disease. Indeed, peptide based caspase inhibitors prevent neuronal loss in animal models of head injury and stroke, suggesting that these compounds may be the forerunners of non-peptide small molecules that halt the apoptotic process implicated in these neurodegenerative disorders. The present review will summarise some of the recent data suggesting that apoptosis inhibitors may become a practical therapeutic approach for both acute and chronic neurodegenerative conditions. PMID- 15359611 TI - [Cerebral transfer and neuroprotection]. AB - In contrast to other vascular beds, the endothelial cells in brain capillaries, which constitute the blood-brain barrier, are sealed together by continuous tight junctions and have little transcellular vesicular transport. In addition to these morphological properties, the presence of specific enzymes and proteins highly restricts the passage of molecules from the blood to the brain. To provide an in vitro system for studying brain capillary functions, we have developed a process of coculture that closely mimics the in vivo situation by culturing brain capillary endothelial cells on one side of a filter and glial cells on the other. In these culture conditions, endothelial cells retain all the endothelial cell markers and the characteristics of the blood-brain barrier, including gamma glutamyl transpeptidase and P-glycoprotein activities. Moreover, the close correlation between the results obtained in vitro with our model and in vivo allows us to conclude that our in vitro blood-brain barrier model is a relevant model for the screening of new molecules to the brain. PMID- 15359612 TI - [The coxibs]. PMID- 15359613 TI - [The controversy surrounding the coxibs: the first report]. PMID- 15359614 TI - [Therapeutic advances with coxibs: few certainties and a lot of doubts]. PMID- 15359615 TI - [The growing area of geriatric medicine]. PMID- 15359616 TI - [Over-consumption of drugs by elderly patients]. AB - Aging is a major factor in the over-consumption of drugs: although individuals aged >65 years account for approximately 15% of the population, almost one-third of all prescriptions are made out for this age group. This over-consumption is responsible for a number of potential hazards, such as iatrogenic pathology, poor compliance and drug interactions; it is also an economic burden. Most studies report an average of five drugs prescribed per individual, regardless of outpatient or inpatient status. Cardiovascular and psychotropic drugs are the most widely prescribed classes. Analgesic and nonsteroidal anti-inflammatory drugs are also very common. These are also the classes responsible for the highest number of adverse events in the elderly. There are numerous reasons for this over-consumption; in some cases, it is attributable to the patient himself (multiple diseases, deterioration of bodily functions, poor compliance, misuse, etc.), while in others, the prescribing physician, family or professional caregivers are at fault. The development of new drugs and the information made available regarding them can also be implicated. Studies comparing actual prescriptions with reference indications have shown that some are inappropriate: one meta-analysis showed that for 21.3% of patients in institutions and 40% of patients in convalescent homes, at least one prescription was inadequate; and in another analysis, the same applied to 12.5% of patients living at home. The potential seriousness of this over-consumption by elderly patients, even more so than in adults, calls for a detailed analysis of the situation and rational, regularly reviewed, realistic prescribing. PMID- 15359618 TI - [Should we treat all the diseases of the elderly?]. AB - The coexistence of several diseases, the loss of autonomy and certain social and behavioural factors lead to polymedication. It is important to reduce the number of drugs prescribed to prevent drug-drug interactions or interactions between drugs and a system weakened by age and comorbiditiy. Because of the limited number of therapeutic trials performed in individuals aged >75 years, clinical consideration based on the consensus of experts is the primary way of establishing priorities and making rational choices. PMID- 15359617 TI - [Is osteoporosis well treated in elderly people?]. AB - Osteoporosis is an increase in bone fragility resulting from a decrease in bone mineral density and abnormalities in the bone micro-architecture. The frequency of osteoporosis is increasing with the aging of patients, and its clinical consequences, vertebral or peripheral fractures, are potentially extremely serious, especially in the very elderly. It is probable that osteoporosis is underdiagnosed--and when it is diagnosed it is often not well treated. It should be realised that a loss in bone mineral density is not the only risk factor for osteoporotic fractures: iatrogenic and environmental factors are also involved, and these are responsive to preventive intervention. Although elderly people usually take multiple drugs, drug treatment, initially calcium and vitamin D, but also bisphosphonates, should also be considered for osteoporosis. PMID- 15359619 TI - [Do premarketing trials help to predict drug-related iatrogenic effects in elderly patients?]. AB - Premarketing trials contribute poorly to predicting drug-related iatrogenic effects in elderly patients. Since their main goal is the demonstration of drug efficacy, these trials are characterised by a simplistic design, they include a limited number of young participants (volunteers only), are of a short duration, and follow a strict protocol. Results of studies in young people cannot be extrapolated to elderly people. Although licensing authorities recommend the recruitment of a meaningful number of elderly people in clinical trials (with an age distribution comparable to that expected when the drug is in routine use), even in trials that are not devoted to geriatric illnesses, elderly people remain substantially under-represented in most instances for methodological reasons (to avoid increased variance introduced by a heterogenous population), safety reasons (at this stage of drug development, it could be deleterious to include patients with comorbid conditions and unfair with regard to the brand image of the products), and ethical reasons (the decision to participate could not be taken by the elderly people alone). Exclusion of elderly participants, who are particularly exposed to drug-related iatrogenic effects, influences the generalisability of study findings. The recruitment of elderly participants, a vulnerable population, is necessary to allow valid conclusions regarding elderly people, recommendations on the appropriate dosage adjustment for elderly individuals, the avoidance of prescribing decisions based on inadequate information (with respect to a more informative summary of the characteristics of the products), and the maximum benefit for elderly people from research. PMID- 15359620 TI - [Particular problems with clinical trials in the fields of cognitive and behavioural diseases]. PMID- 15359621 TI - [What are the limits of the indications and applications of major therapies?: The example of anticancer chemotherapy]. PMID- 15359622 TI - Effects of malnutrition on cytochrome P450 1A2 activity in elderly patients. AB - Little is known of the influence of nutritional status on cytochrome P450 (CYP) 1A2 activity in elderly patients. Thirty elderly institutionalised patients with malnutrition (group A, aged 88 +/- 5 years) and 24 without (group B, aged 81 +/- 9 years) were included. Malnutrition was defined as weight loss of >10% over the previous 6 months and/or a body mass index (BMI) <21 kg/m2 and albuminaemia < or = 32 g/L. CYP1A2 activity was evaluated by the plasma paraxanthine/caffeine (PAX/CAF) metabolic ratio. The plasma PAX/CAF metabolic ratio was similar in both groups regardless of nutritional status (0.34 +/- 0.13 [A] versus 0.30 +/- 0.11 [B]; p = 0.11). The CYP1A2 metabolic ratio was not correlated to either BMI, serum albumin or renal clearance. CYPI A2 activity, as measured by the plasma PAX/CAF ratio, was not influenced by nutritional status in elderly patients. PMID- 15359623 TI - [Drug patents and other ways to protect pharmaceutical research]. AB - Pharmaceutical research constitutes a significant cost for pharmaceutical companies. Because of the importance of the financial investment in research projects, companies must protect their discoveries. There are multiple ways to do this. First, the legal avenue can be divided into three parts: a pharmaceutical company can protect a new drug by a patent, then an additional patent or a secondary patent; moreover, since 1992 in Europe, the pharmaceutical industry has been able to extend a patent by the "Supplementary Protection Certificate" (le Certificat Complementaire de Protection [CCP]). The nonjuridical way is to use the chiral "switch", which can extend patents close to expiring, thus enhancing profitability. PMID- 15359624 TI - [A comparative study of the ocular tolerance after administration of anti allergic eye drops with or without a preservative]. AB - AIM: The aim of the study was to evaluate the occurrence of ocular adverse effects observed after administration of anti-allergic eye drops with and without a preservative in patients with allergic conjunctivitis. METHODS: A total of 3090 patients with allergic conjunctivitis and treated with anti-allergic eye drops were included in an open nonrandomised prospective study by 507 general practitioners located throughout France. The symptoms of discomfort and pain experienced during instillations as well as the characteristics of the patients and of their allergic pathology were recorded. RESULTS: Two groups of patients (eyedrops without preservative [n = 2712] and with preservative [n = 121]) were identified. Sixty percent and 15% of the cases of allergic conjunctivitis were associated with rhinitis and asthma, respectively, and for 70% of the occurrences, an identifiable factor (pollen, dusts, animals etc.) was responsible for the appearance of the symptoms. Compliance was significantly higher for anti allergic eye drops without preservative than for those with a preservative (average number of instillations 3.5 vs 2.9/day, p < 0.001; number of instillations omitted 3.6 vs 4.2, p = 0.01). The proportion of patients experiencing at least one adverse drug reaction was 24% for eye drops with no preservative and 89% for eye drops with a preservative (p < 0.001). The most frequently notified symptom was a sensation of prickling and burning (10% and 47%, respectively, for eye drops with no preservative and eye drops with a preservative; p < 0.001). CONCLUSION: The prescription of eye drops with no preservative allows a significant decrease in ocular adverse drug reactions and a greater acceptance by the patient regarding his/her anti-allergic treatment. PMID- 15359625 TI - [Methcathinone poisoning associated with bromazepam and alcohol]. PMID- 15359626 TI - [Transient carbamazepine overdose after combination with atorvastatin and roxithromycin]. PMID- 15359627 TI - [Biostim (glycoprotein extracted from Klebsiella pneumoniae)-induced Bullous pemphigoid]. PMID- 15359628 TI - Risk assessment in chronic myelomonocytic leukemia (CMML). AB - The clinical course of chronic myelomonocytic leukemia (CMML) is extremely variable, and disease progression can occur at any time from diagnosis. Median survival is about 20 months. About 20% of patients develop acute myeloid leukaemia (AML). Multivariate analyses performed by several groups showed that elevated medullary blast count, low haemoglobin, elevated serum lactate dehydrogenase (LDH), and perhaps an increased lymphocyte count, are the most important independent prognostic parameters, whereas karyotype analysis was not consistently shown to yield additional prognostic information. Applying different scoring systems to 288 CMML patients included in the Dusseldorf MDS Registry, we found that the International Prognostic Scoring System (IPSS) was not useful for defining risk groups in CMML, while the Spanish Score, the modified Bournemouth Score, the Dusseldorf Score, and probably the MDAP Score, identified patient groups differing significantly in survival. These scores should therefore be employed for clinical decision making and for risk stratification in the context of clinical trials. PMID- 15359629 TI - Role of CD47 in erythroid cells and in autoimmunity. AB - The cell surface glycoprotein CD47 (Integrin-associated protein/IAP) was originally identified as a regulator of integrin-dependent responses to extracellular matrix proteins. However, CD47 is ubiquitously expressed, also by cells that do not express integrins. Thus, during the last few years, it has been shown that CD47 has several important functions besides assisting integrin activation. This review will focus on the role of CD47 in erythrocytes. In these cells, CD47 was found to be an important link in the interaction between the band 3 complex and the Rh complex in the maintenance of erythrocyte membrane integrity. CD47 can also function as a marker of self on erythrocytes, and likely also on other cells, by binding to the inhibitory receptor SIRPalpha. In this way, SIRPalpha-expressing cells, like macrophages and dendritic cells, are less likely to phagocytose an autoimmune sensitized cell with CD47 on its surface than a CD47-deficient cell where this inhibitory mechanism will not be engaged. The interaction between CD47 and SIRPalpha seems to be important to limit destruction of host cells in autoimmune diseases like autoimmune hemolytic anemia (AIHA), where macrophages destroy antibody or complement opsonized cells. PMID- 15359630 TI - Ubiquitin ligases in malignant lymphoma. AB - The highly controlled degradation of proteins via the ubiquitin-proteasome pathway represents a key mechanism for cell regulation and homeostasis. Ubiquitin dependent proteolysis, carried out in large part by the E3 ubiquitin ligases, is a critical mode of post-translational modification that is important in regulation of cell cycle progression, signal transduction, gene transcription, antigen receptor signaling, immune response and cell differentiation. Recent studies demonstrate that increasing numbers of proteins with ubiquitin ligase activity are being characterized. Identification and characterization of their substrates indicate that they regulate the turnover of key cell cycle proteins (p27Kip1, p21Cip1, p57Kip2, cyclin E), tumor suppressor proteins (p53, RB), signaling kinases (Src, Zap70, PI-3 kinase), apoptosis regulators (Bcl-2, Bax, Bik) and transcription factors (Myc, NF-kappaB, E1F1), all of which have been implicated in the pathogenesis of malignant lymphoma. Studies to determine the functional role of ubiquitin ligases in the pathogenesis of malignant lymphoma represent potential areas of investigation. PMID- 15359631 TI - The role of NUP98 gene fusions in hematologic malignancy. AB - Chromosomal aberrations occur with great frequency and some specificity in leukemia and other hematologic malignancies. The most common outcome of these rearrangements is the formation of a fusion gene, comprising portions of 2 genes normally present in the cell. These fusion proteins are presumed to be oncogenic; in many cases, animal models have proven them to be oncogenic. One of the most promiscuous fusion partner genes is the newly identified NUP98 gene, located on chromosome 11p15.5, which to date has been observed fused to 15 different fusion partners. NUP98 encodes a 98 kD protein that is an important component of the nuclear pore complex, which mediates nucleo-cytoplasmic transport of protein and RNA. The fusion partners of NUP98 form 2 distinct groups: homeobox genes and non homeobox genes. All NUP98 fusions join the N-terminal GLFG repeats of NUP98 to the C-terminal portion of the partner gene, which, in the case of the homeobox gene partners, includes the homeodomain. Clinical findings are reviewed here, along with the findings of several in vivo and in vitro models have been employed to investigate the mechanisms by which NUP98 fusion genes contribute to the pathogenesis of leukemia. PMID- 15359632 TI - Improved outcome of zygomycosis in patients with hematological diseases? AB - Zygomycosis is an opportunistic fungal infection that is increasingly reported in hematological patients. We describe 2 cases of successfully treated rhino cerebral zygomycosis and give an overview of 120 patients from the literature with underlying hematological or oncological disorders. These data document the improved survival in sinus (15/17 patients surviving) and cutaneous (6/9 patients surviving) disease. Hematological patients with pulmonary (9/30 patients surviving) or disseminated (4/38 patients surviving) zygomycosis still have a poor prognosis. The clinical course of sinus-orbital involvement (4/11 patients surviving) follows sinus-cerebral (2/3 patients surviving) or cerebral (3/6 patients surviving) disease. Besides deoxycholate amphotericin B (AmB) (24/62 patients surviving), patients seem to benefit from liposomal amphotericin B (L AmB) (10/16 patients surviving) or sequential AmB/L-AmB treatment (6/8 patients surviving). Alternative treatment options lead only in a few patients to success. PMID- 15359633 TI - No preferential sensitivity of t(8;21) acute myeloid leukemias to cytosine arabinoside in vitro: is intensity of therapy or high dose Ara-C crucial for response? AB - Acute myeloid leukemia (AML) patients with core binding factor abnormalities [inv(16) or t(8;21)] have a relatively good prognosis, especially patients with inv(16) when treated with high-dose cytosine arabinoside (AraC) containing regimens, whereas in the case of t(8;21) evidences in favor of such regimen are contrasting. We previously demonstrated that blast cells from inv(16)-positive AML patients are characterized by an increased sensitivity to AraC with higher incorporation of 3H AraC into DNA and the increase of induced apoptosis in vitro. In the present study we tested the sensitivity of leukemic cells from 15 t(8;21) positive AML patients to AraC and compared it with the results obtained from cells of 74 patients with inv(16), "intermediate" or "unfavourable" karyotype at diagnosis (for a total of 89 patients). The incorporation of 3H AraC into DNA in cells with t(8;21) was significantly lower than in cells with inv(16) (P = 0.02) or normal karyotype (P = 0.04). Interestingly, the incorporation of the drug into DNA in t(8;21) cells was similar to those with "unfavourable" karyotype. Furthermore, AraC induced apoptosis in t(8;21)-positive AML cells was not increased. These data suggest that the mechanism of response to chemotherapy for t(8;21)-positive cells is probably different then in AML cells with inv(16), underlining the possible importance for patients carrying the t(8;21) of repeated high-dose regimens and not necessarily of high-dose AraC based ones. PMID- 15359634 TI - More on geographic hematology: the breakpoint cluster regions of the PML/RARalpha fusion gene in Mexican Mestizo patients with promyelocytic leukemia are different from those in Caucasians. AB - Acute promyelocytic leukemia is characterized the PML/RARalpha chimeric fusion gene, transcript and protein; the breakpoint cluster regions (bcr) in the PML gene may occur in 3 different sites: Intron 6 (bcr1), exon 6 (bcr2) or intron 3 (bcr3). In a 10-year period in a single institution, we studied prospectively the breakpoint cluster regions of the PML/RARalpha fusion gene in 43 Mexican Mestizo patients with APL, and found that the bcr1 represented 62.7%, the bcr2 9.3% whereas the bcr3 27.9%. The prevalence of the bcr1 subtype is significantly higher than that informed in Caucasians and similar to that in Asians; these data are consonant with those described in other Latin-American patients with APL. Since other Asian genetic markers have been found in the Indian component of the Mexican mestizos, it is possible that the Asian immigration into the Americas through the strait of Behring 12,000 years ago may account for a possible genetic susceptibility to suffer certain forms of APL. PMID- 15359635 TI - Bone marrow microvessel density and its prognostic significance in AML. AB - Microvessel density reportedly is increased in various hematologic disorders including acute lymphatic and myeloid leukemias. In these patients the bone marrow microvessel density (BM-MVD) appears to be associated with an unfavorable prognosis. In the present study, we have retrospectively analyzed the BM-MVD (at diagnosis) in 31 patients with acute myeloid leukemia (AML) (median age: 38 years; range: 21-53 years; f:m-ratio: 1:1,4) who underwent conventional chemotherapy and consecutive allogeneic bone marrow transplantation (BMT). The median BM-MVD at diagnosis was 30/mm2 (range: 17-48/mm2) and thus was significantly higher compared to controls (n = 9; BM-MVD: median 7/mm2, range 2 11/mm2; P < 0.05). In patients who failed to achieve a complete remission (CR) in response to induction chemotherapy, the BM-MVD was significantly higher (median: 41.5/mm2) at diagnosis than in patients who entered CR (median: 28.5/mm2, P < 0.05). In addition, patients with high BM-MVD ( > 30 mm2) had a significantly shorter overall survival compared to patients with a lower BM-MVD ( < 30 mm2, P < 0.05). Moreover, patients with a high BM-MVD ( > 30 mm2) were found to have a significantly higher risk of relapse (P < 0.05). In 4 patients in whom a continuous complete remission was documented after BMT, the BM-MVD levels were analyzed at diagnosis as well as between day + 80 and day + 100 after BMT. In all 4 patients, the BM-MVD was found to decrease in response to BMT until day 100 (P < 0.05). Together, our data suggest that the BM-MVD could be a prognostic parameter concerning survival in patients with AML undergoing allogeneic BMT. PMID- 15359636 TI - Distribution and prognosis of WHO lymphoma subtypes in Taiwan reveals a low incidence of germinal-center derived tumors. AB - To assess the distribution of lymphomas in Taiwan according to the WHO (World Health Organization) classification, 175 recently diagnosed cases of malignant lymphomas were studied and the clinicopathologic data were analyzed. B-cell lymphomas accounted for 57.1% of cases, T-cell lymphomas 38.9%, and Hodgkin's lymphoma 4%. Extranodal lymphomas predominated (55.4%). The most common subtype of B-cell lymphoma was diffuse large B-cell lymphoma (33.1%). All tumor types believed to be derived from germinal center (GC) B-cells including follicular lymphoma (4.6%), Burkitt lymphoma (1.7%), Hodgkin lymphoma (4.0%), and GC-like diffuse large B-cell lymphoma (as defined by combined expression of bc1-6 and CD10) were rather uncommon as compared to frequencies seen in series from Western countries. The common T-cell lymphomas included nasal and extranasal NK/T cell lymphoma (7.4%), mycosis fungoides (7.4%), and unspecified peripheral T-cell lymphoma (6.9%). Adult T-cell leukemia/lymphoma was very uncommon and accounts for only 0.6%. The proportional increase in T-cell lymphomas that were unrelated to type I human T-cell lymphotropic virus (HTLV-1) may be linked to differential Epstein-Barr virus (EBV) oncogenesis. The survival data revealed that mantle cell lymphoma, NK/T-cell lymphoma, unspecified peripheral T-cell lymphoma, and subcutaneous panniculitis-like T-cell lymphoma had an aggressive course. Our results confirm the utility of the WHO classification scheme for prognostic stratification and further highlight the distinctive distribution pattern of malignant lymphoma in Taiwan including the higher relative incidence of T cell lymphomas and the rarity of germinal center-derived B-cell tumors. PMID- 15359637 TI - Adjuvant radiotherapy in stage IV diffuse large cell lymphoma improves outcome. AB - The role of adjuvant radiotherapy to sites of nodal bulky disease in patients with aggressive diffuse large cell lymphoma (DLCL), and stage IV remain undefined. We began a prospective controlled clinical trial to evaluate impact in event free survival (EFS) and overall survival (OS) in a large cohort of patients with a longer follow-up. Between 1989 and 1995; 341 patients with aggressive DLCL and presence of nodal bulky disease (tumor mass > 10 cm) in pathological proven complete response after intensive chemotherapy were randomized to received either radiotherapy (involved fields, 40 Gy) or not. The 5-year EFS and OS in radiated patients were respectively: 82% (95% Confidence interval (CI): 70-89%) and 87% (95% 80-99%), that were statistically significant to control group: 55% (41-64%) (P < 0.001) and 66% (95% CI: 51-73%) (P < 0.01) respectively. Radiotherapy was well tolerated, acute toxicity was mild and until now late toxicity did not appear. The use of adjuvant radiotherapy improve EFS and OS and probably the possibility of cure in patients diffuse large cell lymphoma with worse prognostic factors. Thus, we felt that adjuvant radiotherapy will be considered as part of the initial treatment in this setting of patients. PMID- 15359638 TI - Outcome of elderly patients with aggressive Non-Hodgkin's lymphoma refractory to or relapsing after first-line CHOP or CHOP-like chemotherapy: a low probability of cure. AB - We retrospectively evaluated the outcome of 94 consecutive elderly patients treated at our center for an aggressive lymphoma without a low-grade component. Median survival was 26 months and 5-year overall survival was 39% (27-50%). We then evaluated the outcome of patients refractory to or relapsing after CHOP or CHOP-like chemotherapy. Twenty patients were refractory to first-line therapy and only 1/20 is alive with active lymphoma. Eight patients achieved a partial response and only 3 maintained the partial response while the other 5 patients died. Only 2 of the 27 patients who relapsed after a first complete remission achieved a second sustained complete remission. This study suggests that conventional-dose second-line chemotherapy yields disappointing results in elderly patients with aggressive lymphomas. PMID- 15359640 TI - Chromosomal instability and ATR amplification gene in patients with persistent and polyclonal B-cell lymphocytosis (PPBL). AB - Forty-three patients with persistent and polyclonal B-cell lymphocytosis (PPBL) were studied. The PPBL diagnosis was based on the presence of a polyclonal lymphocytosis and the detection of binucleated lymphoid cells on peripheral blood examination. In order to define the cytogenetic profile in these patients, conventional cytogenetic analysis and fluorescence in situ hybridization were performed at diagnosis in all patients and also at follow-up in 10 patients. When excluding + i(3q) and PCC, chromosomal instability is a common occurrence in PPBL and is characterized by other independent clonal abnormalities, del(6q), + der(8) or + 8, polyploid karyotype, structural changes, aneuploidy and/or non clonal chromosomal aberrations with either loss or more frequently gain of chromosomes. These data show the presence of a chromosomal instability in 67.5% of PPBL patients. Finally, ATR amplification was detected by hybridization with BAC probe 26217 in + i(3q) positive metaphase cells. No ATR deletion was observed in the + i(3q) negative B-cells. As the natural history of PPBL remains unclear, it is necessary to diagnose PPBL patients and useful to recommend a careful and continued long follow-up in all PPBL patients. PMID- 15359639 TI - Cyclooxygenase-2 (Cox-2) expression in lymphomas. AB - Lymphoma is a malign disease of the lymphoid system. A variety of risk factors have been described in pathogenesis of disease. We investigated the role of Cyclooxygenase-2 (Cox-2) in malign lymphomas. A total of 52 patients who were admitted to the Oncology Unit of Mersin University with histologically diagnosed lymphoma were enrolled to this study. Ten of the patients had Hodgkin's disease (HD), and 42 had non-Hodgkin's lymphoma (NHL). An immunuhistochemical method was used for Cox-2 expression. Cox-2 expression was detected in 24 of the 42 patients (57%) with NHL, and it was found in seven of the 10 patients (70%) with HD. The mean patient age expressing Cox-2 was 50.2+/-16.6 years and 48.0+/-15.5 years for patients without Cox-2 expression. This difference was not statistically significant (P = 0.660). The overall survival of Cox-2-positive patients was less than for those without Cox-2 expression but the difference was not significant statistically (16.4+/-11.4 vs. 14.7+/-8.2 months, respectively, P = 0.552) in NHL. There was a correlation between Cox-2 and stage of disease. As the stage increased the Cox-2 expression increased (P = 0.037) in NHL. The complete response rate to therapy was significantly higher in Cox-2-negative patients than the Cox-2-positive group (70.6% vs. 20.8%, respectively, P = 0.001) in NHL. There was no correlation between Cox-2 expression and IPI score, extranodal involvement, tumor grade, and B symptoms. Our findings demonstrate that there is a clinical correlation between the Cox-2 expression and prognostic factors in lymphoma patients. The combination of Cox-2 inhibitors with standard chemotherapeutics may enhance the potential of treatment options for malign lymphomas. PMID- 15359641 TI - Enoxaparin can be used safely in patients with severe thrombocytopenia due to intensive chemotherapy regimens. AB - Treatment with intensive chemotherapy regimens is frequently complicated by severe thrombocytopenia. During the period of severe thrombocytopenia, anticoagulant treatment is not uncommonly indicated for thromboembolic events or thromboprophylaxis in these patients. We report 10 hematological patients treated with intensive chemotherapy protocols that were anticoagulated with enoxaparin for catheter related central venous thrombosis and thromboprophylaxis. During the period of severe thrombocytopenia the dosages of enoxaparin were reduced and no major bleeding occurred. Based on our experience we suggest that reduced dosages of low molecular weight heparins may be used relatively safely during transient severe thrombocytopenia. PMID- 15359642 TI - In multiple myeloma clonotypic CD38- /CD19+ / CD27+ memory B cells recirculate through bone marrow, peripheral blood and lymph nodes. AB - It is believed that myeloma cells are derived from a germinal center (GC) or post GC B cell. The GC B cell can differentiate into both a memory B cell and a plasma cell (PC). In this study, we investigated the recirculating potential of memory B cells clonally related to the myeloma PC (termed clonotypic). The V(H)DJ(H) immunoglobulin gene rearrangement of the myeloma clone was identified for 10 myeloma patients and allele-specific oligonucleotides (ASO) IgH RT-PCR assays were designed for each patient. Memory B cells (CD38- /CD19+ /CD27+) and their subsets defined by the monoclonal antibodies CD62L, CCR6, CXCR4, CXCR5, CCR7 were flow-sorted as single cells and analyzed by ASO RT-PCR analysis. In addition, aspirated peripheral lymph nodes (PLN) of 7 myeloma patients in complete or partial remission were analyzed for the presence of clonotypic cells. Circulating clonotypic memory B cells were identified in PBMNC of 7/10 patients and both CD62L positive and negative clonotypic memory B cells were identified. Furthermore, comparable frequencies of clonotypic cells were found in the CCR6 +/ and CXCR4 +/- memory B cell subsets, whereas all clonotypic memory and later stage B cells were CXCR5 positive. In accordance with their immunophenotype, clonotypic memory B-cells were identified in peripheral blood, bone marrow and PLNs. Clonotypic memory B-cells were present in the majority of myeloma patients and seem to have the same diverse recirculating/homing capacity as normal memory B cells. PMID- 15359643 TI - Generation of dendritic cells from positively selected CD14+ monocytes for anti tumor immunotherapy. AB - Peripheral blood CD14+ monocytes from multiple myeloma (MM) patients can be induced to differentiate into fully functional, mature, CD83+ dendritic cells (DCs) which are highly efficient in priming autologous T lymphocytes in response to the patient-specific tumor idiotype (Id). We have recently scaled up our manufacturing protocol for application in a phase I-II clinical trial of anti-Id vaccination with DCs in MM patients. Elegible patients received a series of by monthly immunizations consisting of three subcutaneous and two intravenous injections of Id-keyhole limpet hemocyanin (KLH)-pulsed DCs (5 x -, 10 x -, 50 x 10(6) cells and 10 x -, 50 x 10(6) cells, respectively). To generate DCs, monocytes were labeled with clinical grade anti-CD14 conjugates and positively selected by immunomagnetic separation. Cells were then cultured, according to Good Manufacturing Practice guidelines, in FCS-free medium in cell culture bags, and differentiated to DCs with GM-CSF plus IL-4 followed by TNF-alpha or, more recently, by a cocktail of IL-1beta, IL-6, TNF-alpha and prostaglandin-E2. Before maturation, Mo-DCs were pulsed with the autologous Id as whole protein or Id (VDJ)-derived HLA class I restricted peptides. Ten MM patients, who had been treated with two courses of high-dose chemotherapy with peripheral blood stem cell support, entered into the clinical study. CD14+ monocytes were enriched from 16.1+/-5.7% to 95.5+/-3.2% (recovery 67.9+/-15%, viability > 97%). After cell culture, phenotypic analysis showed that 89.6+/-6.6% of the cells were mature DCs. We obtained 2.89+/-1 x 10(8) DCs/leukapheresis which represented 24.5+/-9% of the initial number of CD14+ cells. Notably, the cytokine cocktail induced a significantly higher percentage and yield (31+/-10.9 of initial CD14+ cells) of DCs than TNF-alpha alone, secretion of larger amounts of IL-12, potent stimulatory activity on allogeneic and autologous T cells. Storage in liquid nitrogen did not modify the phenotype or functional characteristics of pre-loaded DCs. The recovery of thawed, viable DCs, was 78+/-10%. Thus, positive selection of CD14+ monocytes allows the generation of a uniform population of mature pre loaded DCs which can be cryopreserved with no effects on phenotype and function and are suitable for clinical trials. Based on these results, a DCs-based phase II trial of anti-Id vaccination with VDJ-derived HLA class I-restricted peptides and KLH is underway for lymphoma patients. PMID- 15359644 TI - Upon drug-induced apoptosis in lymphoma cells X-linked inhibitor of apoptosis (XIAP) translocates from the cytosol to the nucleus. AB - The X-linked inhibitor of apoptosis (XIAP) and cellular inhibitor of apoptosis-1 (cIAP-1) are emerging as versatile proteins in programmed cell death with a scope of possible functions reaching far beyond their well known inhibitory effects on caspases. We previously demonstrated that the ability of drugs to modify expression and cleavage of the IAPs are crucial for the synergistic effects achieved by the combinations of different cytotoxic drugs employed to treat malignant lymphomas. In order to more clearly assess the underlying molecular mechanisms, we here evaluated the consequences of drug-induced apoptosis on the localization and aggregation of XIAP and cIAP-1. The influence of drug-induced apoptosis on localization of IAPs was investigated using immunofluorescence microscopy as well as western blot analysis. Apoptosis was induced by chemotherapeutic drugs with different modes of action (bendamustine, cladribine, fludarabine, doxorubicin and mitoxantrone) and assessed by flow-cytometry using Annexin V. We demonstrate that XIAP and cIAP-1 are downregulated and/or cleaved in a dose-dependent manner upon treatment with a variety of anti-cancer drugs. Moreover we provide evidence that in the context of drug-induced apoptosis XIAP, its BIR3-RING cleavage product and cIAP-1 undergo an extensive change of subcellular localization. Immunofluorescence microscopy reveals that XIAP, in contrast to cIAP-1, is located in discrete cytosolic protein aggregates and-upon induction of apoptosis with cytotoxic drugs--redistributes into large nuclear inclusions. This translocation of XIAP and its BIR3-RING cleavage product from the cytosol into the nucleus is confirmed by cell fractionation and western blot analyses. Of note, in this experimental setting putative interaction partners of XIAP-such as Apaf-1, caspase-3 and -7--do not co-localize with XIAP. These results imply a new unknown function of XIAP and its BIR3-RING fragment in the nucleus in the context of drug-induced apoptosis. The localization of cIAP-1 in mitochondria and its liberation from these indicate a profoundly different function of this protein despite its similar modular structure to XIAP. PMID- 15359645 TI - Apoptosis susceptibility and cell-cycle distribution in cells from myelodysplastic syndrome patients: modulatory in-vitro effects of G-CSF and interferon-alpha. AB - Susceptibility to apoptosis varies in different forms of myelodysplastic syndromes (MDS). Our in vitro study aimed at better defining the cell kinetic profile by investigating whether G-CSF and interferon-alpha (IFNalpha) were capable of controling apoptotic/proliferative mechanisms in RAEB as well as in RAEB-t forms. Apoptosis and cell-cycle distribution were measured in mononuclear and in CD34+ cells from bone marrow samples of 27 MDS patients with RAEB (n = 15) and RAEB-t (n = 12). In selected samples, the in vitro influence of G-CSF and lymphoblastoid (Ly)-IFNalpha on the apoptotic susceptibility and on the cell kinetics of the above MDS populations was evaluated. RAEB samples showed a significantly greater apoptosis than RAEB-t ones, both in mononuclear cells (14.76%+/-8.73 vs. 5.95%+/-3.88, P= 0.0058) and in CD34+ cells (24.66%+/-16.08 vs. 3.96%+/-2.57, P = 0.0007). Short-term cell culture in the presence of G-CSF reduced apoptosis in CD34+ cells in all 4 RAEB samples tested (39.1%+/-40.7 vs. 21.0%+/-23.5, P = n.s.); the percentage of cells in S-phase significantly increased in 3/4 samples (19.90%+/-4.40 vs. 32.40%+/-7.85, P = 0.03). Ly-IFNalpha protected CD34+ cells from apoptosis in 3/4 RAEB samples (25.7%+/-8.06 vs. 10.9%+/-8.8, P = n.s.), but did not modulate cell-cycle distribution. G-CSF and Ly-IFNalpha failed to affect apoptosis and proliferation in RAEB-t. These observations indicate that in RAEB forms increased apoptosis can be efficiently counteracted in most of the samples by both G-CSF and Ly-IFNalpha, suggesting that only in these forms a retained regulatory mechanism on the apoptotic/ proliferative balance may allow therapeutic intervention with apoptotic regulators. PMID- 15359646 TI - In the erythroleukemic cell line HEL Prostate-apoptosis-response-gene-4 (par-4) fails to down-regulate Bcl-2 and to promote apoptosis. AB - In a variety of malignant cells Prostate-apoptosis-response-gene-4 (Par-4) exhibits a pro-apoptotic influence sensitizing these cells to apoptosis-inducing agents by downregulating expression of Bcl-2. Considering the crucial role of Bcl 2 in the development of chemoresistance of acute myeloid leukemia (AML) cells, we here assessed the potential of Par-4 to down-regulate Bcl-2 and to induce apoptosis in the erythroleukemic cell line HEL. Testing a potential pro-apoptotic role of Par-4 upon incubation with various conventional chemotherapeutic drugs, novel agents such as the signal transduction inhibitor STI 571 and the histone deacetylase (HDAC)- inhibitor trichostatin A (TSA), as well as with the experimental substances Fas and TRAIL, we provide evidence that in the erythroleukemic cell line HEL expression of Par-4 is not sufficient to sensitize to any of these pro-apoptotic stimuli. We further demonstrate that--in contrast to previous reports in non-AML cells--Par-4 expression in HEL cells leads to an upregulation of Bcl-2. Moreover, Par-4-positive HEL cells exhibit a decreased level of the proapoptotic protein Bax as compared to Par-4- negative cells. In addition, Par-4 increases the expression of Daxx--whose downregulation is associated with augmented chemosensitivity--as well as expression of the procaspases-8, -9 and -10, whereas the levels of the procaspases-3 and -7 remain unaltered. In conclusion we here demonstrate that in the erythroleukemic cell line HEL--in contrast to other cell types Par-4 fails to promote apoptosis and outline the underlying molecular mechanisms. PMID- 15359647 TI - Detection of trisomy 8 in donor-derived Ph- cells in a patient with Ph+ chronic myeloid leukemia successfully treated with Imatinib (STI571) in relapse after allogeneic transplantation. AB - Recent data suggest that STI571 (Imatinib) induces complete cytogenetic responses in patients with chronic myeloid leukemia (CML) who relapse after allogeneic stem cell transplantation (SCT). However, little is known about molecular responses to STI571 and the duration of leukemia-free survival in these patients. We report on a 43 year old female patient who presented with a relapse from Ph+ CML in December 2000. Five years earlier she had received an SCT from an unrelated male donor in accelerated phase. At the time of relapse, she presented with marked leukocytosis (89,000 microl) and 10% blasts. In December 2000, therapy with Imatinib was started. After 3 months, the karyotype showed an XY, with trisomy 8 in about 50% of all metaphases, but without evidence of residual Ph+ cells. Moreover, in response to Imatinib, BCR/ABL transcripts decreased and were no longer detectable after 6 months. After a total observation period of 36 months, the patient is still in complete cytogenetic and molecular remission without signs of occurrence of a donor-type hematopoietic neoplasm or CML relapse. These data suggest that Imatinib is a useful agent for long term treatment of relapsed CML after allogeneic SCT. PMID- 15359648 TI - Primary gastric mantle cell lymphoma in a patient with long standing history of Crohn's disease. AB - The stomach is the most common site of primary extranodal lymphoma. Virtually all cases are of B-cell lineage, including extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT lymphoma) and diffuse large B-cell lymphomas. While secondary gastric involvement from nodal mantle cell lymphoma (MCL) or in the course of primary intestinal MCL (lymphomatous polyposis) have been described, primary gastric MCL has not been reported so far. A 74-year-old man with a 14 year-history of Crohn's disease was admitted at a general hospital due to epigastric pain refractory to therapy with proton-pump inhibitors. Endoscopy disclosed a large polypoid tumor with an ulcerated surface at the greater curvature of the gastric antrum. Endosonography demonstrated the tumor to be limited to the stomach with only local lymph node involvement. Histology of gastric biopsies revealed a dense atypical lymphoid infiltrate composed of small to medium sized cells with slightly irregular nuclear contours. Immunohistochemichally, the cells were positive for CD20, CD79a, CD43 and cyclin D1, but negative for CD3, CD5 and bcl-6. They stained for IgM and showed lambda light chain restriction. Fluorescent in situ hybridisation studies showed the presence of the t(11;14) characteristic for MCL. No further evidence of lymphoma was found on extensive clinical staging. Following chemotherapy the patient is disease free at 24 months after diagnosis. This is the first case of a primary localized gastric MCL. The lack of CD5 expression underscores the importance of performing thorough immunohistochemical studies, particularly to exclude MALT lymphoma. PMID- 15359649 TI - Primary follicular lymphoma of the small intestine. AB - While nodal follicular lymphoma is prevalent in western countries, primary extranodal presentation is rare. We present the clinico-pathological and radiologic features of a patient with primary follicular lymphoma of the small intestine presenting with intestinal obstruction. This is followed by the discussion on the frequency and staging systems for primary gastrointestinal lymphomas, and the relevance of monoclonal anti-CD20 antibody therapy. PMID- 15359650 TI - Extranodal NK/T-cell lymphoma mimicking cellulitis. AB - NK/T-cell lymphoma is difficult to diagnose because there is no characteristic cytology to help the diagnosis in tissue sections, particularly when there is polymorphic cellular infiltration in the early stage of the disease. However, the nasal type of extranodal NK/T-cell lymphoma has a characteristic histologic pattern, which is angiocentric, angioinvasive and angiodestructive. Therefore, many cases of this tumor may show extensive necrosis that mimics infectious process. Furthermore, because the immunosuppressive status of these patients, they may, in fact, have superimposed infections. We are reporting a case that presented as cellulitis and only after careful examination with immunohistochemistry that a correct diagnosis of extranodal NK/T-cell lymphoma, nasal type, was established. Since this lymphoma is incurable and immunophenotyping is instrumental for the diagnosis and prediction of the prognosis, a high index of suspicion for this tumor is needed when an angiocentric lesion is found in the midline of the head and neck region, and a thorough immunohistological study should always be conducted in these cases. PMID- 15359651 TI - Enteropathy-type T-cell lymphoma expressing NK-cell intraepithelial lymphocyte (NK-IEL) phenotype. AB - Enteropathy-type T-cell lymphoma (ETL) is an intraepithelial T-lymphocyte (T-IEL) tumor. The tumor cells are usually CD3+, CD4-, CD8+, and contain cytotoxic granule associated proteins. We report on a CD3-negative CD56-positive enteropathy-associated lymphoma (ETL). This is the first case report of CD3 negative, CD56-positive, CD94-negative, and CD161-positive ETL. ETL cells originate from intraepithelial T-lymphocytes of the intestine. CD3-negative intraepithelial lymphocytes are known as natural killer (NK)-IELs. The phenotype of NK-IELs is also CD3-negative, CD56-positive, CD94-negative, and CD161 positive, while most normal NK cells express CD56 and CD94. CD3-negative lymphoma cells in this report also expressed CD56 and CD161, but not CD94. Because Southern blotting analysis showed a rearrangement of T-cell receptor (TCR) Cbeta in this case, the tumor is classified as an ETL. Based on the findings, NK-IELs may originate from T-cells, not NK-cells. PMID- 15359652 TI - Granulocyte colony stimulating factor-producing multiple myeloma associated with neutrophilia. AB - We report a case of IgG-kappa multiple myeloma associated with neutrophilia (WBC 31,300/microl, neutrophil 90.5%). Interestingly, the serum level of granulocyte colony stimulating factor (G-CSF) in this patient was elevated to 1,500 pg/ml (normal range: 5.78-27.5). Plasma cells were 35% in the bone marrow and were strongly stained with anti-G-CSF antibody. To directly study the production of G CSF from plasma cells in this patient, CD138 positive plasma cells were purified from bone marrow of multiple myeloma patients by magnetic sorting. The expression of G-CSF mRNA was observed in CD138 positive plasma cells from this myeloma patient with neutrophilia by RT-PCR. In contrast, the expression of G-CSF mRNA was not detected in CD138 positive plasma cells from the other multiple myeloma patients without neutrophilia and 4 human myeloma cell lines (HS-Sultan, IM9, RPMI8226, U266) by RT-PCR. After the CD138 positive plasma cells were cultured in vitro for 48 h, the production of G-CSF protein was confirmed (71.8 pg/ml) in the supernatant by ELISA. These results indicated plasma cells of this myeloma patient directly produced G-CSF and that this was the primary cause of neutrophilia. PMID- 15359653 TI - Presentation of extramedullary acute myelogenous leukemia as bilateral testicular masses: response to non-myeloablative allogeneic transplantation. AB - We present an unusual case of extramedullary acute myelogenous leukemia (AML) presenting as bilateral testicular masses. The patient subsequently developed diffuse skin lesions and bone marrow involvement. Although he had only a partial response to intensive chemotherapy, the patient obtained a complete remission after non-myeloablative allogeneic transplantation. PMID- 15359654 TI - Four-year survival after trans-jugular intrahepatic porto-systemic shunt for veno occlusive disease following autologous bone marrow transplantation. AB - Severe hepatic veno-occlusive disease is still a potentially lethal complication after bone marrow transplantation. We here report the case of a patient who developed liver veno-occlusive disease with severe hemodynamic dysfunction and was successfully treated by means of a trans-jugular intrahepatic portosystemic shunt. After three years, he is still disease-free with a functioning shunt and a normal laboratory liver profile. A trans-jugular intrahepatic portosystemic shunt is a treatment option that has been used in very few patients affected by hepatic veno-occlusive disease; its indications and timing remain to be established. PMID- 15359655 TI - Guillain-Barre syndrome complicating mobilization therapy in a case of B-cell chronic lymphocytic leukemia. AB - We report a case of Guillain-Barre Syndrome (GBS) which appeared after mobilization therapy in a patient with B-cell chronic lymphocytic leukemia (B CLL). After obtaining a partial remission with four cycles of fludarabine at standard dose, the patient underwent to high-dose Cytoxan in order to mobilize CD34+ hematopoietic progenitor cells. During neutropenia the patient experienced fever of unknown origin (FUO) and subsequently developed GBS with normalization of his neurologic condition after 2 months. It is possible that a viral-induced activation of an antigen-specific T and B-cell clone caused a local inflammation and toxicity of Schwann cells with demyelination and axonal damage with a self limited course. PMID- 15359656 TI - Teaching cases from the Royal Marsden and St Mary's Hospitals, case 26: refractory microcytic anaemia. PMID- 15359657 TI - Intractable autoimmune hemolytic anemia in B cell chronic lymphocytic leukemia resolved by Rituximab. PMID- 15359658 TI - COX-2 inhibitory treatment in chronic lymphocytic leukemia: a preliminary clinical study. PMID- 15359659 TI - Effect of DCEP mobilizing regimen in in vivo purging of PBSC harvests in multiple myeloma. PMID- 15359660 TI - [Age influence in otoacoustic emissions for hearing loss screening in infants]. AB - OBJECTIVE: To determine the most favorable age for detection of otoacoustic emissions in newborns and for repeated testing. METHODS: Observational, retrospective, descriptive study in 2,567 newborns. RESULTS: The incidence of any degree of hearing loss was 7 per thousand newborns. It was proportionately higher in the group that did not have otologic risk factors The distribution of otoacoustic emissions by age groups followed a significant linear trend in the first month of life. The time lapse to obtain a positive result on the second otoacoustic emission test was 6 days from the first one. CONCLUSIONS: Otoacoustic emission screening should be performed in all newborns as late as possible after birth (from the first 48 hours after birth), but before hospital discharge for the test to be effective and efficient. A repeat test, if required, must be performed at least six days after failing the first one. PMID- 15359661 TI - [Auditory abilities in children with cleft lip and/or palate according to Fisher's]. AB - AIM: Patients with cleft palate and/or lip have similar behaviors to those with auditory processing disorders (APD), then the aim of this study was to investigate auditory abilities in the population with cleft lip/palate. MATERIAL AND METHODS: 100 children between 7 and 12 years old with cleft and 65 between 6 and 10 years old without it. Conventional audiological evaluations such as pure tone, word discrimination audiometry and acoustic immittance measures. A questionnaire (Fisher) was given to parents, with 25 items about behaviors found in children with APD. RESULTS: 100% of children with cleft lip/palate showed some indicative behaviors of APD. CONCLUSION: There was significant occurrence of suggestive behaviors of APD, what points out the importance of inclusion of auditory processing tests in evaluation of children with cleft lip/palate. PMID- 15359662 TI - [The implementation of endoscopic sinus surgery in Spain]. AB - The purpose of this study is to find out more about the implementation of functional endoscopic sinus surgery (FESS) in our country. To do that, we designed a survey which was sent to 160 public hospitals (June 2002). We received the answers of 111 hospitals. 82.9% of hospitals and 58% of surgeons performed FESS, with some differences among autonomic regions. The percentage of surgeons who performed FESS was higher in small hospitals and their mean experience time was 6.2 years. We consider the implementation of endoscopic sinus surgery very high, this can reflect that there are evident advantages for those who specialists who use it. PMID- 15359663 TI - [Endonasal and endocanalicular dacryocystorhinostomy by diode laser. Preliminary results]. AB - AIM OF THE STUDY: To describe the surgical technique and to evaluate the clinical results after having performed the transcanalicular and endocanalicular dacryocystorhinostomies by diode laser, including the advantages and limits of this technique. METHODS: 34 were performed by diode laser in patients with clinical history of epiphora, with or without mucopurulent secretion, for nasolacrimal duct obstruction. The study was prospective, interventional, non randomized and non comparative. Diode laser was used to realize vaporization of lacrimal sac, osteotomy and vaporization with coagulation of nasal mucosa. The mean of surgical time was 15 minutes (range 7 to 29 minutes). Bicanalicular intubation was performed with a silicone tube and prolene filament for two months in all cases. Postsurgical follow-up was between 4 and 11 months. The degree of epiphora was evaluated by the Munk scale and lacrimal permeability was evaluated by endoscopic functional staining test in all cases. RESULTS: Out of the 34 DCR EDN+ENC that were performed, 32 cases (94.11%) remain asymptomatic. Two of them (5.88%) required endonasal dacryocystorhinostomies by drilling, because the bony perforation was impossible to achieve by laser fiber. Two cases (5.88%) presented fibrosis and lacrimal and lower canaliculi obstruction, without epiphora because the superior canaliculi was permeable. CONCLUSION: Endonasal and endocanalicular dacryocystorhinostomy technique performed by diode laser is a valid method. It does not cause cutaneous scarring, it decreases thermic canalicular damage, it respects the lacrimal pump, it minimizes pain and bleeding, it needs less surgical time and it has turned into an out-patient procedure with a minimal surgical and postsurgical morbility. PMID- 15359664 TI - [Cordectomy by laryngofissure. Review of the period 1978-1998]. AB - OBJECTIVE: The early glottic carcinoma can be approached by different therapeutic options. We analyze the results obtained by means of cordectomy opened up by laryngofissure. MATERIAL AND METHOD: We design a descriptive retrospective study including 75 patients with glottic carcinoma T1N0M0 handled by means of cordectomy by laryngofissure (72 males and 3 females). The anaesthetic technique was locoregional and sedation. Tumor T1a, 62 patients (82.66%); T1b, 13 patients (17.33%). RESULTS: The specific survival rate at three, five and ten years was 100, 96.8 and 85.2% respectively; the global survival was 98.66, 90.47 and 52.17% respectively to three, five and ten years. There were ten recurrences (13.3%). Following treatment, 50% of the relapsed patients died. The probability of survival free of illness to three, five and ten years was considered in 0.9322, 0.9172 and 0.7093 respectively. CONCLUSIONS: This technique means good control of the illness and technical easiness. When doing it without a tracheostomy it is tolerated better and it generates lower costs per patient. PMID- 15359665 TI - [Palpation and CT to evaluate the cervical adenopathies in head and neck tumours]. AB - Lymphatic metastasis is an important prognostic factor in patients with head and neck squamous cell carcinomas. Diagnostic evaluation and treatment of this adenopathies is very important. We can use physical examination, computered tomography (CT), magnetic resonance, ultrasound... but none of these give us a 100% security. This study evaluates the accuracy of physical examination and CT in detecting cervical lymph nodes. 120 neck dissections were performed after palpation and CT of 72 patients with head and neck carcinoma. Sensitivity of palpation was 51.7 and specificity 96.7. CTs sensitivity was 65 and specificity 86.7. Both, physical examination and palpation have a high number of mistakes evaluating cervical nodes. N0 necks are still a problem for Head and Neck Surgeons. PMID- 15359666 TI - [Our current results in the surgical treatment of hyperparathyroidism]. AB - We are reporting a descriptive study with our current results in the surgery of hyperparathyroidism. On 14 years experience we have operated 124 patients by performing 122 parathyroidectomies. A 74.2% were primary hyperparathyroidism; a 21.7%, secondary and 4.1% tertiary ones. Among other parameters we have analyzed the clinical symptoms, complementary explorations, surgical technique, anatomopathologic result (AP), location for the removal glands, thyroid pathology associated and complications. Our percentage of success was 96%. We only have registered one recurrent palsy after exposing 190 nerves and 30 hypocalcemias (27 were transitory and 3 permanent). Finally, a literature review has been carried out in order to compare our data with other published series. PMID- 15359667 TI - [Nasal angiofibroma in a 60 year-old female. A clinical case and literature review]. AB - We are reporting an atypical and very rare clinical case of nasal angiofibroma in a 60 year-old female who presented with recurrent epistaxis and nasal blockage. The tumour was located on the right inferior turbinate C.T scans., M.R.I. and Angiography were required as complementary studies. After trying without success a previous arterial embolization, the patient underwent surgery by Functional Endoscopic Nasal Surgery (F.E.N.S.) with removal of the mass in the right nasal fossa. The anatomopathologic result confirmed the histology of angiofibroma and the clinical evolution was satisfactory. Angiofibromas in women and in general in adults are exceptional as we have verified through a literature review. PMID- 15359668 TI - [Incompetent tracheoesophageal fistula following phonatory puncture. Surgical resolution]. AB - Large tracheoesophageal fistulas (TEF) following a total laryngectomy are rare. Most often are associated with the creation of a surgical speech fistula or puncture. We describe the surgical technique used in 3 patients with large TEF after a total laryngectomy and the creation of a speech fistula with a voice prosthesis, followed by radiation therapy. Pertinent literature is reviewed. PMID- 15359669 TI - Safety symbol comprehension: effects of symbol type, familiarity, and age. AB - A new procedure for evaluating symbol comprehension, the phrase generation procedure, was assessed with 52 younger and 52 older adults. Participants generated as many phrases as came to mind when viewing 40 different safety symbols (hazard alerting, mandatory action, prohibition, and information symbols). Symbol familiarity was also assessed. Comprehension rates for both groups were lower than the 85% level recommended by the American National Standards Institute. Moreover, older participants' comprehension was significantly worse than younger participants', and the older adults also generated significantly fewer phrases. Generally, prohibition symbols were comprehended best and hazard alerting symbols worst. In addition, symbol familiarity was positively correlated with symbol comprehension. These findings indicate that important safety information depicted on signs and household products may be misunderstood if presented in symbolic form. Furthermore, certain types of symbols may be better understood (e.g., prohibition symbols) than other types (e.g., hazard alerting symbols) by both younger and older individuals. These findings signify the utility of the phrase generation procedure as a method for evaluating symbol comprehension, particularly when it is not possible or desirable to provide contextual information. Actual or potential applications of this research include using the phrase generation approach to identify poorly comprehended symbols, including identification of critical confusions that may arise when processing symbolic information. PMID- 15359670 TI - Conceptual issues in the study of dynamic hazard warnings. AB - This paper presents a conceptual analysis of dynamic hazard warning systems. The normative aspects of responses to warnings are analyzed, and a distinction is made between two forms of responses to a warning system, referred to as compliance and reliance. Determinants of the responses to warnings are identified, and they are broadly classified into normative, task, and operator factors. Existing research on warnings and automation is assessed in view of this conceptual framework, and directions for future research are discussed. Some implications of this analysis for practitioners, designers, and researchers are indicated. Actual or potential applications of this research include recommendations for the analysis, design, and study of dynamic warning systems. PMID- 15359671 TI - Verification of the change blindness phenomenon while managing critical events on a combat information display. AB - Change blindness occurs when humans are unable to detect significant changes in objects and scenes after their attention is momentarily diverted. Because change blindness is relevant in many applied settings, the current study investigated the phenomenon in the context of tasks performed by naval command and control system personnel. Operators of such systems are often heavily loaded with concurrent visual search, situation assessment, voice communications, and control display manipulation tasks at large, physically dispersed tactical situation displays. As the operators' attention shifts from one display to another, it creates an opportunity for changes to occur on unattended screens with potentially negative consequences. Our results show that on a display containing 8 objects of interest, considerable change blindness was demonstrated in that participants required 2 or more selections to correctly identify a changed object on nearly 1/3 of the test trials. Further, operator performance on 15% of the trials was equivalent to randomly guessing with replacement after making 3 incorrect selections. This research underscores the need for developing effective countermeasures to the change blindness phenomenon. Actual or potential uses of this research include interface design of computer workstations for military, nuclear power industry, air traffic control, crisis response center, and hospital emergency room applications. PMID- 15359672 TI - Effects of sensory modality and task duration on performance, workload, and stress in sustained attention. AB - The workload and stress associated with a 40-min vigilance task were examined under conditions wherein observers monitored an auditory or a visual display for changes in signal duration. Global workload scores fell in the midrange of the NASA Task Load Index, with scores on the Frustration subscale increasing linearly over time. These effects were unrelated to the sensory modality of signals. However, sensory modality was a significant moderator variable for stress. Observers became more stressed over time as indexed by responses to the Dundee Stress State Questionnaire, with evidence of recovery in the auditory but not the visual condition toward the end of the watch. This result and the finding that signal detection accuracy - although equated for difficulty under alerted conditions - favored the auditory mode, indicate that display modality and time on task should be considered carefully in the design of operations requiring sustained attention in order to enhance performance and reduce stress. Actual or potential applications of this research include domains in which monitoring is a crucial part, such as baggage screening, security operations, medical monitoring, and power plant operations. PMID- 15359673 TI - Productivity and ergonomic investigation of bent-handle pliers. AB - Awkward wrist posture is generally considered an occupational risk factor for hand/wrist disorders, leading to the ergonomic design principle of "bend the tool, not the wrist." Sixteen participants performed a computer jumper installation task and a simple assembly task while productivity, wrist posture, and shoulder posture were measured. The work surface orientation (vertical and 45 degrees) and the level of constraint placed on the user (constrained grip and unconstrained grip) were also varied. The results indicate that the beneficial effects of the bent-handle pliers are task dependent. In the computer jumper task the bent-handle pliers resulted in 5.3% faster task performance, whereas in the assembly task performance was 4.9% faster with the straight-handle pliers. The bent-handle pliers reduced shoulder deviations by 50% in the jumper installation task, and ulnar deviation was reduced by 12% and 22% for the jumper installation task and the assembly task, respectively (all significant at p < .05). However, allowing participants to hold the pliers in a grip configuration of their choosing (unconstrained technique) often reduced these postural benefits. In applying these results to workplace design activities, one should recognize that the ergonomic utility of bent-handle pliers can be considerable but that the 3-D kinematics characteristics of the task must be considered. PMID- 15359674 TI - Grip force vectors for varying handle diameters and hand sizes. AB - Grip force was measured along two orthogonal axes and vector summed. Sixty-one participants recruited from a manufacturing facility (29 men and 32 women) grasped instrumented cylinders (2.54, 3.81, 5.08, 6.35, and 7.62 cm diameter) using a maximal voluntary power grip. Two orthogonal force measurements relative to the third metacarpal were resolved into a magnitude and corresponding angle. On average, magnitude increased 34.8 N as handle diameter increased from 2.54 cm to 3.81 cm, and then monotonically declined 103.8 N as the handle diameter increased to 7.62 cm. The average direction monotonically decreased from 59.2 degrees to 37.7 degrees as handle diameter decreased from the largest to the smallest. When the diameter was smallest, the greatest force component, Fx (168.6 N), was in the direction where the fingertips opposed the palm. Conversely, when the diameter was largest, the smallest component, Fx (77.7 N), was in the same direction. These values are averaged for the left and right hand. The angle for the largest diameter increased with increasing hand size. These relationships should be useful for the design of handles that require gripping in specific directions, such as for hand tools and controls. Actual or potential applications of this research include the design of handles that require gripping in specific directions, such as for hand tools and controls, that reduce effort, and that prevent fatigue and overexertion. PMID- 15359675 TI - Trapezius muscle motor unit activity in symptomatic participants during finger tapping using properly and improperly adjusted desks. AB - Work-related musculoskeletal disorders in the neck-shoulder area and upper extremities are common among computer users, especially women. We compared temporal changes of motor unit (MU) activation in the trapezius muscle during finger tapping using both appropriate and inappropriate ergonomic desk adjustments. Sixteen intensive and nonintensive computer users with either moderate or severe musculoskeletal disorders participated in the study. Six channel intramuscular electromyographic (EMG) signals and 2-channel surface EMG were recorded from 2 positions of the trapezius muscle. A statistically significant increase in activity was observed with a desk adjusted 5 cm higher than appropriate and was attributable mainly to increased duration of MU activity. Participants with severe symptoms activated more MUs, and these were also active longer. In women, on average, MUs were active nearly twice as long as in men during the same tapping task. This study demonstrates that it is possible to evaluate ergonomic topics on the MU level and that incorrectly adjusted office equipment, in addition to motor demands imposed by the work task, results in prolonged activity of MUs. A potential application of this research is an increased awareness that certain individuals who work with incorrectly adjusted office equipment may be at greater risk of developing work-related musculoskeletal disorders. PMID- 15359676 TI - Right of way in the sky: two problems in aircraft self-separation and the auction based solution. AB - There has been a growing movement to give commercial airliners more freedom in choosing their routes and responsibility for detecting and avoiding conflicts. These "free flight" concepts must contain new rules for assigning right of way in potential conflict situations. To evaluate the effect of prospective rules, the current paper derives the expected response of agents who exhibit different levels of sophistication. Traditional game theoretic analysis is used to derive the behavior of rational agents. Computer simulations are used to predict the behavior of boundedly rational reinforcement learners. The results reveal that several seemingly reasonable, straightforward right-of-way rules might lead to undesirable outcomes. These problematic results are robust to the assumed level of rationality. It is shown that these problems can be alleviated by using auctions to resolve competition for right of way. Actual or potential applications of this research include the usage of second price auctions to address right-of-way and similar conflicts. PMID- 15359677 TI - Remembrance of cases past: who remembers what, when confronting critical flight events? AB - Recent "naturalistic" theories of decision making emphasize the role of stored prior experiences or cases as a guide to current action. However, there is little empirical evidence on the role that case-based remindings play in real-life decision making. The present study utilized a Web-based survey to collect data about the role of prior cases in pilot decision making about critical flight events. Results showed that more than half of the 1081 pilots who responded could provide details about utilizing a previous case in responding to a critical flight event. These events were most likely to involve weather or equipment failure. The cases were found to be useful in situational assessment rather than option evaluation. The use of cases increased with age and experience. Data obtained from a concurrent conventional survey showed broadly similar results. The implications of these results are that case-based remindings play an important role in expert pilot decision making and that a training system that incorporates case-based learning would be a potentially useful means of improving pilot decision making. Actual or potential applications of this research include the development of case-based training systems to enhance flight training. PMID- 15359678 TI - Evaluation of a computerized aid for creating human behavioral representations of human-computer interaction. AB - The research reported herein presents the results of an empirical evaluation that focused on the accuracy and reliability of cognitive models created using a computerized tool: the cognitive analysis tool for human-computer interaction (CAT-HCI). A sample of participants, expert in interacting with a newly developed tactical display for the U.S. Army's Bradley Fighting Vehicle, individually modeled their knowledge of 4 specific tasks employing the CAT-HCI tool. Measures of the accuracy and consistency of task models created by these task domain experts using the tool were compared with task models created by a double expert. The findings indicated a high degree of consistency and accuracy between the different "single experts" in the task domain in terms of the resultant models generated using the tool. Actual or potential applications of this research include assessing human-computer interaction complexity, determining the productivity of human-computer interfaces, and analyzing an interface design to determine whether methods can be automated. PMID- 15359679 TI - Foot shape modeling. AB - This study is an attempt to show how a "standard" foot can be parameterized using foot length, foot width, foot height, and a measure of foot curvature so that foot shape can be predicted using these simple anthropometric measures. The prediction model was generated using 40 Hong Kong Chinese men, and the model was validated using a different group of 25 Hong Kong Chinese men. The results show that each individual foot shape may be predicted to a mean accuracy of 2.1 mm for the left foot and 2.4 mm for the right foot. Application of this research includes the potential design and development of custom footwear without the necessity of expensive 3-D scanning of feet. PMID- 15359680 TI - Minimal instrumentation may compromise failure diagnosis with an ecological interface. AB - Interfaces designed according to ecological interface design (EID) display higher order relations and properties of a work domain so that adaptive operator problem solving can be better supported under unanticipated system conditions. Previous empirical studies of EID have assumed that the raw data required to derive and communicate higher-order information would be available and reliable. The present research examines the relative advantages of an EID interface over a conventional piping-and-instrumentation diagram (PID) when instrumentation is maximally or only minimally adequate. Results show an interaction between interface and the adequacy of the instrumentation. Failure diagnosis performance with the EID interface with maximally adequate instrumentation is best overall. Performance with the EID interface drops more drastically from maximally to minimally adequate instrumentation than does performance with the PID interface, to the point where the EID interface with minimally adequate instrumentation supports nonsignificantly worse performance than does the equivalent PID interface. Actual or potential applications of this research include design of instrumentation and displays for complex industrial processes. PMID- 15359681 TI - Engaging in activities involving information technology: dimensions, modes, and flow. AB - An engagement mode involves a subject (e.g., a user of information technology, or IT) who is engaged in an activity with an object in a certain manner (the mode). The purpose of this study is to develop a general model of engagement modes that may be used for understanding how IT-related activities are shaped by properties of the user and the IT object. A questionnaire involving items on IT engagement and the experience of flow was administered to 300 participants. The results supported an engagement mode (EM) model involving 5 different engagement modes (enjoying/acceptance, ambition/curiosity, avoidance/hesitation, frustration/ anxiety, and efficiency/productivity) characterized on 3 dimensions (evaluation of object, locus of control between subject and object, and intrinsic or extrinsic focus of motivation). The flow experience follows from a balance between enjoying/ acceptance and efficiency/productivity propelled by ambition/curiosity. The EM model could provide a platform for considering how IT users, IT applications, and IT environments should work together to yield both enjoyment and efficiency. Actual or potential applications of this research include designing IT training programs on different levels of specificity. PMID- 15359682 TI - The size-weight illusion in team lifting. AB - The purpose of these studies was to examine whether or not there is a size-weight illusion when participants lift containers as a team. In Experiment 1, teams of participants lifted a set of 16 containers that varied in mass and size and reported their perceptions of heaviness and volume. In Experiment 2, participants lifted the same containers individually. A size-weight illusion was demonstrated in each experiment: Reports of perceived heaviness decreased substantially as the volumes of the containers increased for both styles of lifting. However, both the magnitude of the illusion and mean perceived heaviness were greater in the team lift. Actual or potential applications of this research include the development of safe lifting guidelines for team lifting. PMID- 15359683 TI - Imperfect in-vehicle collision avoidance warning systems can aid drivers. AB - An experiment was conducted to determine the effects of an in-vehicle collision avoidance warning system (IVCAWS) on driver performance. A driving simulator was driven by 135 licensed drivers. Of these, 120 received alerts from the IVCAWS when their headway to a lead car was less than 2 s, and the other 15 (the control group) received no alerts. Drivers received varied alert interfaces: auditory, visual, and multimodal. The system had varied levels of reliability, determined by both false alarm rate and failure of the IVCAWS to alert to short headway. Results indicated that the IVCAWS led to safer (longer) headway maintenance. High false alarm rates induced drivers to slow down unnecessarily; large numbers of missed alerts did not have any significant impact on drivers. Driver acceptance of the system was mixed. Interface played a role in driver reliance on the system, with the multimodal interfaces generating least reliance. Actual or potential applications of this research include IVCAWS interface selection for greater system efficacy and user acceptance and the advisability of implementation, even of imperfect systems, for drivers who seek to maintain a safer headway. PMID- 15359684 TI - Synchronization of traffic signals as a means of reducing red-light running. AB - The goal of this study was to determine if traffic signals that are synchronized along a route are associated with fewer red-light violations than traffic signals that are not synchronized. A total 3600 cycles of traffic signals at 12 intersections along 2 major urban arteries were observed. Synchronized intersections were effective in reducing the likelihood of red-light running (RLR) by (a) providing fewer opportunities than nonsynchronized intersections for RLR and (b) having a lower rate of RLR relative to the number of opportunities. After adjustment for the number of opportunities, the odds of entering the intersections in red in synchronized intersections were nearly 1/7 the odds of RLR in nonsynchronized intersections. Congestion reduced the effectiveness of synchronized intersections relative to nonsynchronized intersections. Male drivers were slightly more likely to run red lights than female drivers, and the effects of synchronization were fairly constant across age, gender, and the presence or absence of passengers. Actual or potential applications of this research involve signal synchronization to reduce aggressive driving in general and RLR in particular. PMID- 15359685 TI - [Essay on the history of the Department of Human Anatomy of Iaroslavl State Medical Academy]. AB - The article reviews the history of the Department of Human Anatomy of Yaroslavl State Medical Academy since the moment of its foundation to the present time (1944-2004). The contribution of the noted scientists that headed the Department is described, with special regard to the progress in science and the educational process. PMID- 15359686 TI - [Principles of morphogenesis of the visceral innervation during different stages of human postnatal development]. AB - This atricle presents the results of the studies of the neurovascular complexes in intestinal arteries, spleen, ovary, testis, thyroid gland, liver, kidney, heart, brain, common carotid, femoral and humeral arteries and great saphenous vein, obtained from neonates, mature and old humans. Within each age group, the normative parameters of adventitial plexuses are established according to the number of macro-microscopic and microscopic nerve bundles and their absolute and relative content of thin, intermediate and thick myelinated nerve fibers (MNFs) (Weigert-Pal's staining), relative density of acetylcholinesterase-positive fibers in nerve plexuses (Koelle-Gomori's and Karnovsky-Ruts methods), presence of destructive changes in MNFs (Bielschowsky-Gross's method). Comparison of adventitial plexuses construction in different arteries suggests the regularity in the development of nervous connections of visceral organs in human postnatal ontogenesis according to the following periods: productive, stabilization and involution. Productive period is characterized by the increase in the number of nerves and their MNF content, formation of heterogeneous population of MNFs belonging to different morpho-functional classes. In the period of relative stabilization no changes were found in the number of nerves and their MNF content, while after the age of 60 massive MNF destruction was detected. Organ specificity and an individual status of the nervous connections of the organs studied during postnatal human life was demonstrated. PMID- 15359687 TI - [Histoenzymochemical characteristic of neuromuscular synapse in skeletal muscles]. AB - The paper summarizes the results of longstanding research on the construction of AChE-positive zone in neuromuscular synapse (NMS) in relation to the metabolic profile, muscle fibre diameter and function of the muscles of the head, neck, anterior and posterior extremities of mature albino rat. Comparison of cross sectional areas and forms of NMS region indicates that the values of these parameters are significantly associated with the muscle fibre diameter. Other muscle characteristics are less important for the construction of AChE-positive zone of NMS in mature rat. PMID- 15359688 TI - [Transformations of enzyme-active zones in neuromuscular synapses of some skeletal muscles of different origin during postnatal ontogenesis]. AB - The aim of this study was to detect the age-dependent transformations of the enzyme-active zones (EAZ) in neuromuscular synapses (NMSs) of skeletal muscles of different origin. As the objects of study, m. triceps brachii, m. rectus dorsalis oculi, m. sternocleidomastoideus and m. pectoralis major were used, that were taken from the rats aged 3 and 21 days, 1, 3 and 6 months, 1, 1,5 and 2 years. The histochemical method modified by Nikolayev and Shilkin (1983) was used. It was demonstrated that starting from the moment of birth and up to the 1st month of life EAZ in NMSs are mainly represented by simple homogeneous structures. From 1st to 6th month the region of NMS acquired the features which were peculiar to particular muscle. After 6 months and up to 2 years, natural age-related degenerative and regenerative processes were observed in a muscle. The transformations of NMS region in the muscles of varoius origin appeared at different time. PMID- 15359689 TI - [Development of neuromuscular synapse in the skeletal muscle after chemical desympathization]. AB - The purpose of this investigation was the study of cholinesterase (ChE)-positive zones of neuromuscular synapse (NMS) in gastrocnemius and plantaris muscles of albino rats after guanethidine sympathectomy. Qualitative and morphometric characteristics of ChE-positive zones were studied, which were demonstrated using a modified thioacetic acid method in 72 control and experimental animals aged from 5 days to 3 months. It was found that in both gastrocnemius and plantaris muscles the growth of muscle fiber diameter and section area in the region of NMS was delayed after desympathization. In gastrocnemius muscle, smaller fiber diameter persisited up to the age of 3 months, while in plantaris muscle it was larger than in control animals of the same age. The differences in dimensional parameters of NMS sections were leveled off by the age of 3 months because of their accelerated growth. Desympathization resulted in the premature appearance of dystrophic changes in MNS region as compared to those developing in control animals, in which they are the features of involution processes. PMID- 15359690 TI - [Calcium ions localization in mixed synapses of mauthner cells after exposure to substances changing the gap junction conductivity]. AB - Using the pyroantimonate method, the distribution of calcium ions in Mauthner cell mixed synapses was studied following their treatment with substances changing gap junction (GJ) synapse electrotonic conductivity. Ecdyson, the drug known to increase GJ conductivity, induced the appearance of calcium pyroantimonate precipitates covering the whole postsynaptic area of GJs, making them highly asymmetric. Formation of precipitate grains was also observed in the cleft of desmosome-like contacts (DLCs). Chlorpromazine, substance that decreases GJ conductivity, induced the formation of precipitates in GJ cleft and on both pre- and postsynaptic sides of their surfaces. No precipitates were seen in DLC cleft. These results show that ecdyson acts as a substance selectively increasing the GJ conductivity with no effect on DLC function. Chlorpromazine has dual effect, by blocking the communication through both GJ and DLC. Thus, using chemicals that change GJ permeability, the possibility of their interaction with actin-containing structures which also participate in the electrotonic signal transfer, should be taken into consideration. PMID- 15359691 TI - [Intramural nervous apparatus of jejunum and rectum in experimental portal hypertension]. AB - Using histological and electron microscopical methods, the state of intramural nervous structures of jejunum and rectum was studied in 92 dogs with experimental portal hypertension. Three phases of changes of portal pressure were detected and its influence upon the intramural nervous apparatus was observed. In phase 1 (first 4-5 days after the surgery) the reactive changes of the nervous apparatus were shown that were more pronounced in jejunum. In phase 2 (day 5 to 2.5 months) the portal pressure was shown to drop; this was accompanied by some signs of regeneration in jejunum and by an aggravation of destructive processes in rectum. In phase 3 of the repeated rise of portal pressure (2.5 to 6.5 months after the surgery), destructive changes in the nervous apparatus were demonstrated that were similar to those found in phase 1. However, the differences in the reaction of nervous structures in different layers of intestinal tube and the heterogeneity of the changes of the nervous apparatus in jejunum and rectum were absent. The association of time of appearance and of severity of structural changes with the elevation of portal pressure suggests the significant role of vascular factor in the morphogenesis of alterations observed in phase 3 of hemodynamic changes. PMID- 15359692 TI - [Age-dependent changes of morphometric and histochemical characteristics of neurocytes in different ganglia of albino rats]. AB - The aim of this study was to obtain the normative data on the age-dependent transformation of morphometric and histochemical characteristics of neurocytes in different ganglia in albino rats. Cell cross-sectional area, activities of cholinesterase (demonstrated with thioacetic acid method) monoamine oxidase (demonstrated with Glenner method) were measured in neurocytes of stellate, spinal, trigeminal and gastric ganglia in rats aged 2 to 360 days. Measurements were made with the help of "Bioscan" videoanalyzer. Informational analysis was used for the evaluation of the degree of maturation of neurocyte systems. General features, age- and organ-related peculiarities of morphometric and enzyme histochemical characteristics were established for neurocytes of different ganglia, as well as a heterochronism of their definitive state attainment. The time of stabilization for neurocytes of stellate and I thoracic spinal ganglia was the age of 60 days, for those of trigeminal ganglion and intramural gastric ganglia -90 and 120 days, respectively. By this time, neurocyte systems turned from a determined state into a probabilistic-determined one, this transformation being considered as a population stabilization. PMID- 15359693 TI - [Survival and phenotypic characterisctic of axotomized dorsal root ganglion neurons]. AB - Using indirect inmunohistochemical method, the expression of Bcl-XL and Bax, anti and proapoptotic proteins of Bcl-2 family, as well as of cytokine IL-1beta were studied to demonstrate the role of these substances in apoptosis regulation of sensory neurons of different subpopulations after the severance of their peripheral processes. For comparison of the capacity of these neurons to survive after central and peripheral axotomy, the expression of high-molecular component of neurofilament triplet NF200 and isolectin B4 (IB4) binding were studied. At day 30 after central axotomy, the total number of neurons in LIV-LV ganglia of rat was not changed, but the number of NF200+ neurons was decreased. In mouse Lv dorsal root, proapoptotic BaX protein was demonstrated in the nuclei of 46% of small neurons that accounts for 20% of the total neuronal number in this ganglion. By day 30 after the nerve crush separate Bax expression was found in the nuclei of 30% and in the cytoplasm of 20% of neurons. In intact animals, dorsal root ganglion antiapoptotic Bcl-XL protein was expressed in the cytoplasm of 30% of small neurons, as well as in satellite cells surrounding large and intermediate neurons. At day 30 after the nerve crush Bcl-XL expression in LV ganglion was not detected. IL-1beta was present in the cytoplasm of 17% of neurons belonging predominantly to the subpopulations of large and intermediate neurons. By day 30 after the nerve crush IL-1(+-neurons were not found. PMID- 15359694 TI - [Preganglionic inputs to cat stellate ganglion in postnatal ontogenesis]. AB - Localization of the spinal cord preganglionic neurons projecting to the stellate ganglion, was studied in kittens from the moment of birth till six month of life, using the method based on retrograde axonal transport of horseradish peroxidase. The number of preganglionic neurons and spinal cord segments connected with the stellate ganglion, increased from the moment of birth till 10th postnatal day, decreasing afterwards. The proportion of preganglionic neurons in different nuclei of the spinal cord changed during the development and became comparable to that in adult animals by 6th month. The proportion of neurons in the nucleus intermediolateralis thoracolumbalis pars principalis was found to increase, while the number of neurons in the nucleus intermediolateralis thoracolumbalis pars funicularis and in the spinal cord ventral horns was decreased. PMID- 15359695 TI - [Interleukin-6 in rat pterygopalatine ganglion neurons]. AB - Rat pterygopalatine ganglion was studied by indirect immunofluorescent method to detect and to localize the neurons expressing IL-6. The highest concentration of IL-6-immunopositive cells was found in the cranial part of the ganglion, while ganglion medial surface and caudal part contained single cells demonstrating bright fluorescence; less common groups of immunopositive neurons, organized in chains, were found in the middle part of the ganglion near its lateral border that is in contact with maxillar nerve. PMID- 15359696 TI - [Afferent innervation in the cervical region of the esophagus and trachea in early postnatal ontogenesis]. AB - Method of retrograde axonal transport of horseradish peroxidase was used to study the afferent innervation of trachea and esophagus cervical portion in the neonatal, 10-, 20- and 30-days old kittens. Labeled neurons in all the animals were localized in the vagal caudal sensory ganglion and CI-CVI spinal ganglia both in right and left sides. The major role in the innervation of trachea and esophagus cervical portion in kittens from the moment of their birth till postnatal day 20, is played by the cells of spinal ganglia, while at day 30 these are neurocytes of the vagal ganglia. No significant differences were found in the number of neurocytes, innervating the trachea and the esophagus. PMID- 15359697 TI - [Morphometric characteristic of some structures of the human brain in normal conditions and in acute ethanol intoxication]. AB - In order to establish the neuro-stromal interrelations in the human brain under normal conditions and after extreme influences, different parts of this organ were studied in 14 persons who died suddenly and in 20 persons who died of ethyl alcohol poisoning. A complex of histological, histochemical, planimetric and morphometric methods were used. The specific area of normal neurocytes was shown to be the largest in the medulla oblongata, while that one of the fibers--in the cerebral cortex. Vascularization of the latter is minimal, being much higher in the medulla oblongata. In alcohol intoxication, the area, occupied by neurocytes, was decreased in the cerebral cortex, thalamus and cerebellum due to the destruction of some portion of these cells. In the medulla oblongata neurocytes appear to be more resistant to ethanol. The capillary diameter in the brain areas studied decreased because of the drop in the tone of cerebral arteries, but the number of these vessels per standard area of section was increased as a result of compensatory opening of reserve capillaries. PMID- 15359698 TI - [Interhemispheric connections of areas 17 and 18 in the cortical columns of cats with unilateral strabismus]. AB - The interhemispheric connections of areas 17 and 18 of the cerebral cortex were investigated in cats with experimental unilateral strabismus. Single cortical columns were microiontophoretically injected with horseradish peroxidase, and retrogradelly labeled cells were demonstrated in the opposite brain hemisphere. After tracer injection in area 18 columns, the labeled callosal cells were located in area 17/18 transitional zone, similar to what was found in normal cats. However, in some cases the expansion of the region of callosally labeled cells distribution, was found. It is proposed that the extent of the region of callosally-connected cells may vary depending on whether the cells receive their input from intact or strabismic eye. PMID- 15359699 TI - [Neuronal damage in the hippocampus of inbred mouse strains in the models of epilepsy induced by kainic acid and pilocarpine]. AB - Elucidation of the mechanisms of neuronal damage is an important task of modem neuroscience and is of paramount importance for medicine. Present work compares two models of excitotoxic neuronal damage induced by kainic acid and pilocarpine, in which inbred C57BL/6J (C57BL) and FVB/NJ (FVB) mice were used. Both models produced higher neuronal damage in FVB although mortality was higher in C57BL. No significant differences between two strains of mice were found in seizures severity. Kainic acid demonstrated greater tropism to hippocampus in comparison with pilocarpine. Hsp-70 and Egr-1 expression was not significantly different in C57BL and FVB. Analysis of the isolated mitochondrial fraction has shown different degree of free radical production in the strains studied, that could be one of the reasons for unequal susceptibility of their neurons to excitotoxic cell death. PMID- 15359700 TI - [Induction of NO-synthase and glial fibrillary acidic protein in astrocytes of the temporal cortex in rats with audiogenic epileptiform reaction]. AB - The localization of NADPH-diaphorase (NADPH-d), inducible NO-synthase (iNOS) and glial fibrillary acidic protein (GFAP) was studied in the astrocytes of the temporal cortex in rats of Krushinsky-Molodkina strain which are genetically prone to audiogenic seizures. The seizure was evoked by thrice-repeated acoustic stimulation. Wistar rats and acoustically untreated seizure-free Krushinsky Molodkina rats were used as a control. The foci of brain damage were consistently found in the neocortex of the animals with audiogenic seizures. Epileptic foci, 300-400 microm in diameter, were localized in layers III-V; they were found to consist of the clusters of NADPH-d-positive astrocytes and to be present in both hemispheres. In the foci of cortical damage astrocytes expressed iNOS and an elevated level of GFAP. The number of GFAP-immunopositive astrocytes in the foci of damage was increased by 25-37% compared to the control and to undamaged areas of the cortex. Astrocyte NOS and GFAP induction found in this work, suggests the participation of glia in compensatory NO-dependent mechanisms, that are formed in the damage foci of neocortex during the audiogenic seizures. PMID- 15359701 TI - [Morphochemical characteristic of rat brain structures after exposure to delta sleep inducing peptide following long-term amphetamine administration]. AB - The aim of this work, that was carried out using Wistar rats, was to characterize the response of neurons of different morphofunctional types to amphetamine administration and to study the possibility of correction of these changes by delta-sleep inducing peptide (DSIP). Single intraperitoneal injection of 60 microg/kg of DSIP following a long term amphetamine administration (2.5 mg/kg for 3 weeks) was shown to result in normalization of brain metabolism, that was disturbed by the drug. The correcting DSIP effect was found in rat brain structures judging by the parameters of the state of proteins in neurons of sensomotor cortex and caudate nucleus and by the activity of enzymes of neurotransmitter metabolism, such as type A and B monoamine oxidases and acetylcholine esterase, that was determined in subfractions of the same brain structures. DSIP modulating effect in phenamine stereotypy supports its role as an adaptogen of intercenter relations in CNS pathology. PMID- 15359702 TI - [Cellular composition of cerebral cortex in rat fetuses exposed to low-dose fractionated radiation]. AB - Using histological and morphometric methods, the peculiarities of sensomotor cortex morphogenesis were studied in rat fetuses subjected to fractionated gamma irradiation in the period from 6th to 18th prenatal days in doses of 5, 25, 50 and 25-75 cGy. It that an Exposure to a fractionated radiation was found to negatively affect stem cell proliferation in the developing cortex and to increase the intensity of cell destruction in proportion to the radiation dose. The absolute number of macroglial cells was increased in all the cellular zones of developing cortex, that could be the result of both reactive stimulation of their production due to functional insufficiency, and of accelerated transformation of radial gliocytes into macroglia. Irradiation with the doses of 5-75 cGy caused the deceleration of neuroblast migration into the primary cortex, as indicated by the reduction of cell number in neural differon in the neocortical anlage and cell number growth in deeper layers of the developing cortex. PMID- 15359703 TI - [Interrelations of morphological parameters in the system of neuroendocrine regulation following longterm morphine administration]. AB - To study the morpho-physiological changes of the nervous and endocrine regulatory systems following long-term influence of morphine hydrochloride, morphometric and correlation analysis of the structural components of the cerebral cortex, hypothalamus, adenohypophysis and thyroid gland was performed. The interrelation between the values of some parameters and the dose of morphine hydrochloride, that caused the tolerance to the drug, was shown. Dose-dependent effect was found in the degree of morphological remodeling of organ components that was demonstrated by the changes in cell and nuclear dimensions, staining properties of cytoplasm, ultrastructural components of thyrocytes, neurons and their processes, hormonal background, the number of the functioning blood capillaries. Interrelations of the morphological parameters are demonstrated by the correlations of different strength and direction. PMID- 15359704 TI - [Morphometric characteristics of microcirculatory bed of olfactory bulbs in the desympathized albino rat]. AB - To detect the effect of postnatal chemical desympathization on morphometric characteristics of the vessels of olfactory bulbs in albino rat early postnatal ontogenesis, the study was performed using the histochemical method for demonstration of alkaline phosphatase activity, which is a marker of vascular endothelium, and vessel diameter and relative density were measured during the first three months of life. Desympathization was induced in rats by daily intraperitoneal injection of 60-70 mg/kg of guanethidine during postnatal days 1 to 30. The vessel diameter was found to increase after day 30 simultaneously with the changes of the vascular wall of vessels (irregular contour and non-uniform vasodilatation, non-uniform decrease in alkaline phosphatase activity). Relative density of the vessels was reduced one month after desympathization. After two months this parameter remained depressed in the peripheral zone of the olfactory bulbs, while in the central zone it was not different from that one in control group. PMID- 15359705 TI - [Morphological characteristics and immunophenotype of dendritic cells generated from monocytes of human peripheral blood]. AB - Morphological features and immunophenotype of mature dendritic cells (DC) generated from the monocytes of healthy donor blood after culture with GM-CSF and IL-4 with the addition of TNF, were studied using light and electron microscopy, as well as flow-cytometry. It was shown that DC were characterized by a number of morphological features such as: large size, eccentrically located nucleus, highly developed system of extensions, large vacuolated cytoplasm, prominent and activated Golgi complex and ribosomal apparatus. Mature DC are characterized by active surface expression of MHC and co-stimulatory molecules (MHC I, MHC II, CD40, CD80, CD86). PMID- 15359706 TI - [Morpho-functional changes of splenic autotransplants at the different time intervals after the operation]. AB - This investigation was aimed at the study of splenic autotransplant cytoarchitectonics and cellular structures functional activity 3-60 days after the operation. The studies were carried out in 70 albino male rats including intact animals (n-10) and rats subjected to splenectomy and subsequent spleen heterotopic autotransplantation (n-60). Cryostat sections of transplants were processed for Falc and Cross luminescent-histochemical methods and were stained with hematoxylin and eosin. Biogenic amines content was measured using LUMAM-4 microscope with FMEL-1A spectrofluorimetric device. The number of luminescent cells in periarterial T-dependent zone was found to increase significantly after 7 days, while the unchanged lymphoid nodules were found in the zone of unaltered tissues. At day 15 "intrafollicular" luminescent granulocytes appeared in the newly formed lymphoid nodules. At days 15 to 45 the increased content of biogenic amines was detected in the "intrafollicular" cells, while the significant rise in the amount of amines was seen in the lymphocytes of nodule T-independent zone. It is suggested that the presence of intact nodules in transplant peripheral zone together with an increase in cell numbers in T-dependent zone, are the key events in the development of immune-correction effect of the transplanted spleen during the first two weeks after the operation. PMID- 15359707 TI - [Cytochemical characteristics of neurons in the posterior amygdala]. AB - This investigation was aimed at the comparative analysis of the intensity of RNA precursor 3H-uridine incorporation in the neurons of nuclear, transitional formations and paleocortical structures of amygdala posterior division. Significant differences were detected in the label incorporation intensity in the neurons of piriform cortex as compared to that one in neurons of the medial portion of posterior cortical nucleus. PMID- 15359708 TI - [Immunocytochemical demonstration of astrocytes in brain sections combined with Nissl staining]. AB - The aim of the present study was to develop an easy and reliable protocol of combined preparation staining, which would unite the advantages of immunocytochemical demonstration of astrocytes with the availability to evaluate functional state of neurons provided by Nissl technique. The presented protocol of paraffin sections processing allows to retain high quality of tissue structure and provides for selective demonstration of astrocytes using the monoclonal antibodies against glial fibrillary acidic protein and contrast Nissl staining of cells. The protocol can be used without any changes for processing of brain sections obtained from the humans and other mammals with the exception of mice and rabbits. PMID- 15359709 TI - [Adrenergic regulation of the structure and function of the parathyroid glands]. AB - In this review the current concepts on the sources of sympathetic innervation of the parathyroid glands (PTG) of mammals are systematized; morphological and histochemical characteristics of intraorganic sympathetic nerves are described. The influence of experimental sympathetic denervation and pharmacological modulation of adrenoreceptors on secretory and proliferative activity of parathyrocytes was analyzed. The main mechanism of influence of catecholamines released from the nerve endings and transported from the blood, on parathyrocytes is their diffuse action on cell receptor apparatus, mostly on beta adrenoreceptors located in plasma membrane. In physiological regeneration conditions, catecholamines are able to increase secretory and mitotic activity of pararthyroid epithelium. In reparative regeneration of the PTG, chemical sympathetic denervation and pharmacological modulation of beta-adrenoreceptors lead to a decrease in the expression and duration of parathyrocyte proliferative response; at the organ level this resulted to in the appearance of the tendency towards development of focal-type proliferative reaction instead of diffuse ones. PMID- 15359710 TI - [On the anatomical terms used for description of brain arteries and discrepancy between them and anatomical nomenclature]. AB - The following terms are used in Russian literature to designate arteries, which carry blood into the brain: brachiocephalic arteries, aortic arch branches, main cerebral arteries (arteries of the head), extra- and intracranial cerebral arteries, precerebral arteries. All terms either have substantial drawbacks or conflict with the adopted anatomical terminology. These discrepancies become far more significant in view of publication of "Terminologia Anatomica" (New York, 1998). We suggest to discuss the possibility to use universal terminology for the designation of the system of the arteries, supplying the brain. We propose to designate truncus brachiocephalicus, common carotid and subclavian arteries, internal carotid and vertebral arteries with their branches including arterial network on the brain surface by the term "the conducting arteries", whereas small branches, which pierce the brain matter are to be named "the penetrating arteries" (T.M. Sundt et al., 1976). Considering the Greek word "encephalon", which means "the brain", all the arteries of the brain could be designated as encephalic, while conducting arteries--as preencephalic, and intracerebral (penetrating) arteries--as intraencephalic. PMID- 15359711 TI - Editorial: Of continuities, beginnings, and generativities. PMID- 15359712 TI - The emergence of "family integrity" in later life. AB - In contrast to the rich and abundant literature on the early stages of the family life cycle, there are relatively few theoretical accounts of family developmental processes in the second half of life. In an effort to address this imbalance, we introduce the concept of "family integrity" to refer to the ultimate, positive outcome of an older adult's developmental striving toward meaning, connection, and continuity within his or her multigenerational family. Subjectively, for the older adult, family integrity may be experienced as a deep and abiding sense of peace and/or satisfaction with his or her multigenerational family relationships, past, present, and future. It is the result of a growing concern for relational closure that typically begins to emerge in mid-life and gains ascendance in later life. Objectively, at the level of observable relational processes, it is manifest in three interrelated competencies of the family as a system: (a) the transformation of relationships over time in a manner that is dynamic and responsive to the changing life cycle needs of individual family members, (b) the resolution or acceptance of past losses or family conflicts, with the dead as well as the living, and (c) the shared creation of meaning by passing on individual and family legacies within and across generations. As a construct, family integrity involves processes at multiple levels of social organization. At the individual level, it refers to an older adult's inner experience of satisfaction or "completeness" in the context of his or her family relationships. At the family level, it refers to observable relational competencies and transactions that contribute directly to an elder's sense of meaning, purpose, and connection with others. Simultaneously, at the level of culture and society, there are values and rituals that influence whether individuals experience meaning and purpose within the multigenerational family. PMID- 15359713 TI - The importance of parents and other caregivers to the resilience of high-risk adolescents. AB - Relationships between 43 high-risk adolescents and their caregivers were examined qualitatively. Parents and other formal and informal caregivers such as youth workers and foster parents were found to exert a large influence on the behaviors that bolster mental health among high-risk youth marginalized by poverty, social stigma, personal and physical characteristics, ethnicity, and poor social or academic performance. Participants' accounts of their intergenerational relationships with caregivers showed that teenagers seek close relationships with adults in order to negotiate for powerful self-constructions as resilient. High risk teens say they want the adults in their lives to serve as an audience in front of whom they can perform the identities they construct both inside and outside their homes. This pattern was evident even among youth who presented as being more peer-than family-oriented. The implications of these findings to interventions with caregivers and teens is discussed. PMID- 15359714 TI - Same-sex couple perceptions of family support: a consensual qualitative study. AB - Few studies have examined the family context in which same-sex couples negotiate their lives and relationships. Consensual qualitative research methods (Hill, Thompson, & Williams, 1997) were used to analyze 14 same-sex couples' conversations about family support. Couples perceived that their families are positively supportive, non-supportive, or ambivalent in their support. These perceptions led to positive or negative emotional reactions in the couple members and to specific coping. The majority of couples perceived that family support (or lack of support) had an effect on the quality of their couple relationship. No general or typical response strategies to lack of family of origin support emerged, suggesting a lack of models or norms for same-sex couples. The implications for psychotherapeutic interventions with same-sex couples are discussed. PMID- 15359715 TI - The practice of community family therapy. AB - This paper presents a summary of the basic theories and methods of Community Family Therapy (CFT), a relatively new therapeutic approach developed in response to the need for effective intervention in treating low-income, urban families. CFT operates outside of the traditional therapeutic box, successfully combining family therapy techniques with developmental and motivational theories, community mental health, social work, economic development, and community mobilization strategies. CFT utilizes a dualistic approach in which both client and therapist become involved with the same three levels of engagement. Specifically, the client strives for: (a) personal and family change and growth-level 1, (b) accessing community resources-level 2, and (c) leadership development and civic action-level 3. Also, CFT calls upon therapists to strive for: (a) personal growth and maturation-level 1, (b) collaboration with community resources for professional support-level 2, and (c) operation as a "citizen therapist," through civic action and volunteer services-level 3. PMID- 15359716 TI - Family-level impact of the CHAMP Family Program: a community collaborative effort to support urban families and reduce youth HIV risk exposure. AB - This article presents family-level results from an ongoing study examining the impact of the CHAMP (Chicago HIV prevention and Adolescent Mental health Project) Family Program, a family-based HIV preventative intervention meant to reduce the amount of time spent in situations of sexual possibility and delay initiation of sexual activity for urban youth in the 4th and 5th grades living in neighborhoods with high rates of HIV infection. The CHAMP Family Program has been developed, delivered, and overseen by a collaborative partnership, consisting of community parents, school staff, community-based agency representatives, and university based researchers. Design of the program was informed by input from this collaborative partnership, child developmental theory of sexual risk, and empirical data gathered from the targeted community. This article presents findings that suggest CHAMP Family Program impact on family communication, family decision-making, and family-level influences hypothesized to be related to later adolescent HIV risk. Implications for future family-based HIV prevention research are discussed here. PMID- 15359717 TI - A pilot study of an intervention for breast cancer survivors and their spouses. AB - Recent studies have shown that interventions that increase breast cancer patients' communication with family members lead to reduced patient distress. In this article, we report on a treatment development and pilot study of an intervention for couples coping with breast cancer. In phase 1 of this study, 10 couples participated in two focus groups that generated ideas and themes for the intervention. In phase 2, we developed and pilot tested our intervention with 48 couples: 12 in a 2-session format, 21 in a 1-session format, and 15 in a non experimental control group. Our response rate shows that breast cancer patients and spouses were willing to participate and that treatment providers were willing to refer patients and their spouses. The 2-session format showed the most promise for producing positive change in mental health functioning and cancer-related stress. PMID- 15359718 TI - Ontogenesis in narrative therapy: a linguistic-semiotic examination of client change. AB - In this article I investigate how the narrative therapy process facilitates client change. The kind of change that I focus on is linguistic-semiotic; that is, how clients develop their meaning potential through language. What I will demonstrate is how an examination of the linguistic-semiotic level provides new insights into narrative therapy's role in endowing clients with the semiotic materials to make new meanings. An examination of six conjoint sessions involving a narrative therapist with one couple revealed that client change or ontogenesis is composed of three semiotic phases. In the first phase of ontogenesis clients display a beginning semiotic repertoire by formulating "extreme case" descriptions of self and other's behaviors. In the second phase clients are scaffolded by therapist's questions and reformulations into construing events as problems and problems as the agents of negative behaviors. In the final phase clients display a development in their semiotic potential. Clients are able to eliminate problems and construe themselves as agents without prior therapist scaffolding. Therefore, in the latter stages of the narrative process clients are able to deploy meanings that have been generated throughout therapy, in order to produce narratives of self agency and self control. PMID- 15359719 TI - Factor structure of the FACES-III in Japanese university students. AB - We performed a confirmatory factor analysis for the Family Adaptability and Cohesion Evaluation Scale (FACES-III) using 3865 university students in Japan. We confirmed two factors as in the original version, but there were slight differences with the original one. Some items were deleted in order to obtain sufficient goodness-of-fit indexes in a series of confirmatory factor analyses. Moreover, items 11 "Our family, changes its way of handling tasks," 20 "Parents and children discuss in our family," and 15 "We shift household responsibilities," which were originally categorized as "adaptability" items, were loaded on the "cohesion" factor. PMID- 15359720 TI - Do we have enough biomarkers? PMID- 15359721 TI - Identifying membrane glycoproteins. PMID- 15359722 TI - Clinical and pharmacogenomic data mining: 2. A simple method for the combination of information from associations and multivariances to facilitate analysis, decision, and design in clinical research and practice. AB - The physician and researcher must ultimately be able to combine qualitative and quantitative features from a variety of combinations of observations on data of many component items (i.e., many dimensions), and hence reach simple conclusions about interpretation, rational courses of action, and design. In the first paper of this series, it was noted that such needs are challenging the classical means of using statistics. Hence, the paper proposed the use of a Generalized Theory of Expected Information or "Zeta Theory". The conjoint event [a,b,c,..] is seen as a rule of association for a,b,c,.. associated with a rule strength I(a;b;c;...) = xi(s,o[a,b,c,..]) - xi (s,e[a,b,c,...]), where xi is the incomplete Zeta Function. Here, o[a,b,c,...] is the observed, and e[a,b,c,..] the expected, frequency of occurrence of conjoint event [a,b,c,...]. The present paper explores how output from this approach might be assembled in a form better suited for decision support. Related to this is the difficulty that the treatment of covariance and multivariance was previously rendered as a "fuzzy association" so that the output would fall into a similar form as the true associations, but this was a somewhat ad hoc approach in which only the final I( ) had any meaning. Users at clinical research sites had subsequently requested an alternative approach in which "effective frequencies" o[ ] and e[ ] calculated from the above variances and used to evaluate I( ) give some intuitive feeling analogous to the association treatment, and this is explored here. Though the present paper is theoretical, real examples are used to illustrate application. One clinical genomic example illustrates experimental design by identifying data which is, or is not, statistically germane to the study. We also report on some impressions based on applying these techniques in studies of real, extensive patient record data which are now emerging, as well as on molecular design data originally studied in part to test the ability to deduce the effects of simple natural patient sequence variations ("SNPs") on patient protein activity. On the basis of these study experiences, methods of rationalizing and condensing the rules implied by associations and variances between data, as well as discussion of the difficulty of what is meant by "condensed", are presented in the Appendix. PMID- 15359723 TI - Multidimensional protein profiling technology and its application to human plasma proteome. AB - In clinical and diagnostic proteomics, it is essential to develop a comprehensive and robust system for proteome analysis. Although multidimensional liquid chromatography/tandem mass spectrometry (LC/MS/MS) systems have been recently developed as powerful tools especially for identification of protein complexes, these systems still some drawbacks in their application to clinical research that requires an analysis of a large number of human samples. Therefore, in this study, we have constructed a technically simple and high throughput protein profiling system comprising a two-dimensional (2D)-LC/MS/MS system which integrates both a strong cation exchange (SCX) chromatography and a microLC/MS/MS system with micro-flowing reversed-phase chromatography. Using the microLC/MS/MS system as the second dimensional chromatography, SCX separation has been optimized as an off-line first dimensional peptide fractionation. To evaluate the performance of the constructed 2D-LC/MS/MS system, the results of detection and identification of proteins were compared using digests mixtures of 6 authentic proteins with those obtained using one-dimensional microLC/MS/MS system. The number of peptide fragments detected and the coverage of protein sequence were found to be more than double through the use of our newly built 2D-LC/MS/MS system. Furthermore, this multidimensional protein profiling system has been applied to plasma proteome in order to examine its feasibility for clinical proteomics. The experimental results revealed the identification of 174 proteins from one serum sample depleted HSA and IgG which corresponds to only 1 microL of plasma, and the total analysis run time was less than half a day, indicating a fairly high possibility of practicing clinical proteomics in a high throughput manner. PMID- 15359724 TI - Liquid chromatography MALDI MS/MS for membrane proteome analysis. AB - Membrane proteins play critical roles in many biological functions and are often the molecular targets for drug discovery. However, their analysis presents a special challenge largely due to their highly hydrophobic nature. We present a surfactant-aided shotgun proteomics approach for membrane proteome analysis. In this approach, membrane proteins were solubilized and digested in the presence of SDS followed by newly developed auto-offline liquid chromatography/matrix assisted laser desorption ionization (LC/MALDI) tandem MS analysis. Because of high tolerance of MALDI to SDS, one-dimensional (1D) LC separation can be combined with MALDI for direct analysis of protein digests containing SDS, without the need for extensive sample cleanup. In addition, the heated droplet interface used in LC/MALDI can work with high flow LC separations, allowing a relatively large amount of protein digest to be used for 1D LC/MALDI which facilitates the detection of low abundance proteins. The proteome identification results obtained by LC/MALDI are compared to the gel electrophoresis/MS method as well as the shotgun proteomics method using 2D LC/electrospray ionization MS. It is demonstrated that, while LC/MALDI provides more extensive proteome coverage compared to the other two methods, these three methods are complementary to each other and a combination of these methods should provide a more comprehensive membrane proteome analysis. PMID- 15359725 TI - Tumor suppressor gene 14-3-3sigma is down-regulated whereas the proto-oncogene translation elongation factor 1delta is up-regulated in non-small cell lung cancers as identified by proteomic profiling. AB - Lung cancer, a leading cause of cancer deaths, consists of two major groups: small cell lung cancer (SCLC) and nonsmall cell lung cancer (NSCLC) with the NSCLC accounting for approximately 75% cases of lung cancers. It has been suggested that molecular changes including overexpression of oncogenes and decreased expression of tumor suppressor genes are responsible for lung carcinogenesis. In this study, we analyzed protein profiles of four different human NSCLC cell lines compared with normal human bronchial epithelial cells using two-dimensional PAGE and MALDI-TOF mass spectrometry. We identified 12 protein spots with different expressions between the normal and cancer cells. Of these proteins, vimentin, cytokeratin 8, YB-1, PCNA, Nm23, hnRNP A2/B1, and HSP90beta were known to be up-regulated in lung cancers, which is consistent with the current study. We also found that the expression of M-type pyruvate kinase is altered in NSCLC likely due to changes in translational control and/or differential phosphorylation of the protein. Interestingly, the expression of the tumor suppressor gene 14-3-3sigma is down-regulated while that of the proto oncogene TEF1delta is up-regulated in NSCLC cells. On the basis of these observations and previous studies, we propose that the altered expression of 14-3 3sigma and TEF1delta may be involved in lung carcinogenesis. PMID- 15359726 TI - Comparison of multiplexed techniques for detection of bacterial and viral proteins. AB - Immobilized antibody microarrays were compared to the Luminex flow cytometry system that utilizes suspensions of polystyrene microbeads covalently coupled with capture antibodies. The two immunoassays were performed for comparison of reproducibility, limits of detection and dynamic range. The Luminex system showed lower limits of detection and increased dynamic range among samples whereas the protein microarrays could be more amenable to miniaturization. Both technologies were capable of sensitive multiplexed detection. PMID- 15359727 TI - New approach for analysis of the phosphotyrosine proteome and its application to the chicken B cell line, DT40. AB - In this study, we have begun to analyze phosphotyrosyl and associated proteins present in a DT40 chicken B cell line overexpressing the nonreceptor protein tyrosine kinase, Syk. An anti-phosphotyrosine antibody was used to select tyrosine-phosphorylated proteins. After tryptic digestion, peptides were subjected to a beta-elimination reaction and phosphotyrosine-containing peptides were enriched via immobilized metal affinity chromatography. Several known substrates and candidate substrates for Syk and the location of 22 tyrosine phosphorylation sites were identified. PMID- 15359728 TI - Statistical and mechanistic approaches to understanding the gas-phase fragmentation behavior of methionine sulfoxide containing peptides. AB - Recently, we carried out a statistical analysis of a 'tryptic' peptide tandem mass spectrometry database in order to identify sequence-dependent patterns for the gas-phase fragmentation behavior of protonated peptide ions, and to improve the models for peptide fragmentation currently incorporated into peptide sequencing and database search algorithms [Kapp, E. A., Schutz, F., Reid, G. E., Eddes, J. S., Moritz, R. L., O'Hair, R. A. J., Speed, T. P. and Simpson, R. J. Anal. Chem. 2003, 75, 6251-6264.]. Here, we have reexamined this database in order to determine the effect of a common post-translational or process induced modification, methionine oxidation, on the appearance and relative abundances of the product ions formed by low energy collision induced dissociation of peptide ions containing this modification. The results from this study indicate that the structurally diagnostic neutral loss of methane sulfenic acid (CH3SOH, 64Da) from the side chain of methionine sulfoxide residues is the dominant fragmentation process for methionine sulfoxide containing peptide ions under conditions of low proton mobility, i.e., when ionizing proton(s) are sequestered at strongly basic amino acids such as arginine, lysine or histidine. The product ion abundances resulting from this neutral loss were found to be approximately 2-fold greater than those resulting from the cleavage C-terminal to aspartic acid, which has previously been shown to be enhanced under the same conditions. In close agreement with these statistical trends, experimental and theoretical studies, employing synthetic "tryptic" peptides and model methionine sulfoxide containing peptide ions, have determined that the mechanism for enhanced methionine sulfoxide side chain cleavage proceeds primarily via a 'charge remote' process. However, the mechanism for dissociation of the side chain for these ions was observed to change as a function of proton mobility. Finally, the transition state barrier for the charge remote side chain cleavage mechanism is predicted to be energetically more favorable than that for charge remote cleavage C-terminal to aspartic acid. PMID- 15359729 TI - Application of peptide LC retention time information in a discriminant function for peptide identification by tandem mass spectrometry. AB - We describe the application of a peptide retention time reversed phase liquid chromatography (RPLC) prediction model previously reported (Petritis et al. Anal. Chem. 2003, 75, 1039) for improved peptide identification. The model uses peptide sequence information to generate a theoretical (predicted) elution time that can be compared with the observed elution time. Using data from a set of known proteins, the retention time parameter was incorporated into a discriminant function for use with tandem mass spectrometry (MS/MS) data analyzed with the peptide/protein identification program SEQUEST. For singly charged ions, the number of confident identifications increased by 12% when the elution time metric is included compared to when mass spectral data is the sole source of information in the context of a Drosophila melanogaster database. A 3-4% improvement was obtained for doubly and triply charged ions for the same biological system. Application to the larger Rattus norvegicus (rat) and human proteome databases resulted in an 8-9% overall increase in the number of confident identifications, when both the discriminant function and elution time are used. The effect of adding "runner-up" hits (peptide matches that are not the highest scoring for a spectra) from SEQUEST is also explored, and we find that the number of confident identifications is further increased by 1% when these hits are also considered. Finally, application of the discriminant functions derived in this work with approximately 2.2 million spectra from over three hundred LC-MS/MS analyses of peptides from human plasma protein resulted in a 16% increase in confident peptide identifications (9022 vs 7779) using elution time information. Further improvements from the use of elution time information can be expected as both the experimental control of elution time reproducibility and the predictive capability are improved. PMID- 15359730 TI - 'Signature sets', minimal fragment sets for identifying protein disulfide structures with cyanylation-based mass mapping methodology. AB - Our cyanylation (CN)-based methodology for determining disulfide structure of cystinyl proteins overcomes the limitations of conventional proteolytic methods. However, the CN-based method has the potential drawback that occasionally some CN induced cleavage fragments may not be detected. We show that CN-based methods can overcome the failure to detect fragments by demonstrating the existence of small 'signature sets' of fragments. The link between signature sets and the robustness of CN-based methodology is validated by two case studies. PMID- 15359731 TI - On invariants of a 2-D proteome map derived from neighborhood graphs. AB - We consider the problem of the construction of invariants for characterization of 2-D maps, such as 2-D proteome maps, 2-D NMR spectral maps, etc., that in addition to facilitating cataloguing such maps, can be used for comparison of maps and numerical evaluation of their degree of similarity. A novel approach, based on the concept that the nearest neighborhood of points (spots) on a map are sufficiently flexible to allow one not only to vary the number of points used for characterization of the map but also the density of information on their relative positions, is put forward. The method is illustrated with the Coomassie brilliant blue stained 2-D gel electrophoresis patterns of the proteomes from liver cells of healthy male Fisher F344 rats and the rats treated with four peroxisome proliferators. PMID- 15359732 TI - Global differential non-gel proteomics by quantitative and stable labeling of tryptic peptides with oxygen-18. AB - We describe a protocol for quantitative labeling of tryptic peptides with oxygen 18. Proteins are first digested in natural water with trypsin, the pH is then lowered to 4.5 and the mixture is dried. Oxygen-18 water is added and two oxygen 18 atoms are incorporated at the peptides' carboxyl termini. Trypsin is finally inactivated by cysteine alkylation under denaturing conditions, which blocks oxygen back-exchange. The general value of this labeling strategy for differential proteomics is illustrated by the analysis and identification of several couples of differently labeled amino terminal peptides isolated from a human platelet proteome by a previously described chromatographic procedure. PMID- 15359733 TI - Characterization of the human salivary basic proline-rich protein complex by a proteomic approach. AB - Thirteen samples of human normal whole saliva were analyzed by RP-HPLC-ESI-MS and MALDI-TOF-MS to investigate the basic proline-rich protein complex. Between known basic-PRPs the P-B, P-C (or IB-8b), P-D (or IB-5), P-E (or IB-9), P-F (or IB-8c), P-H (or IB-4), IB-6, II-2, IB-1, and IB-8a glucosylated were identified, whereas the II-1, IB-7, PA, and D1-A peptides were not detected. Some detected masses not attributable to known basic-PRPs were putatively ascribed to II-2 and IB-1 nonphosphorylated, II-2 and IB-1 missing the C-terminal arginine residue, and the 1-62 fragment of IB-6, named P-J peptide. A correlation matrix analysis revealed a cluster of correlation among all the basic PRPs (apart from the P-B peptide) which is in agreement with their common parotid origin. PMID- 15359734 TI - Improved molecular weight-based processing of intact proteins for interrogation by quadrupole-enhanced FT MS/MS. AB - Complete coverage of protein primary structure is demonstrated for 37 yeast protein forms between 6 and 30 kDa in an improved platform for Top Down mass spectrometry (MS). Tandem mass spectrometry (MS/MS) for protein identification with 100% sequence coverage is achieved in a highly automated fashion with 15-300 fold less sample amounts than an initial report of a proteome fractionation approach employing preparative gel electrophoresis with an acid-labile surfactant to facilitate reversed phase separation in a second dimension. Using a quadrupole enhanced Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FTICRMS) improves the dynamic range for protein detection by approximately 50-fold and MS/MS by approximately 30-fold. The technology development illustrated here typifies an accelerating effort to detect whole proteins in a more general and higher throughput fashion for improved biomarker identification and detection of diverse post-translational modifications. Capillary RPLC is used in both off-line and on-line modes, with one on-line LC/FTMS sample providing 25 observed protein forms from 11 to 22 kDa. PMID- 15359735 TI - Proteolysis and mass spectrometric analysis of an integral membrane: aquaporin 0. AB - Due to hydrophobicity, structural analysis of integral membrane proteins poses a formidable challenge for current mass spectrometry-based proteomics approaches. Herein, we demonstrate results from optimized sample preparation and enzymatic proteolysis procedures for the complete primary structure determination of a targeted integral membrane protein, lens aquaporin 0 (AQP0). Plasma membrane from bovine lens tissue was alkali treated and tryptic digestion was performed in optimized acetonitrile-ammonium bicarbonate solution. Full sequence coverage of AQP0 was observed as tryptic peptides using both matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) and capillary liquid chromatography tandem mass spectrometry (cLC/MS/MS). An amino acid mutation of Thr to Ile/Leu at residue 199 was deduced based on MS/MS results. In a complementary effort to fully sequence the protein, peptic digestion was developed to take advantage of hydrophobic protein solubility in organic acid as well as the decreased activity of pepsin at low pH. Peptic digestion in 10% formic acid (pH 1.2) generated peptides of 500 to 3000 Da and gave 100% sequence coverage by cLC/MS/MS. In addition to post-translational modifications reported previously, a new phosphorylation site at serine 229 and two oxidation sites at tryptophan 202 and 205 were detected on the protein. These methodologies provide complementary detergent- and CNBr-free procedures for detailed analysis of this important membrane channel protein and offer promise for analysis of the integral membrane proteome. PMID- 15359736 TI - Peptide identification using peptide amino acid attribute vectors. AB - We describe the theoretical basis for a peptide identification method wherein peptides are represented as vectors based on their amino acid composition and grouped into clusters. Unknown peptides are identified by finding the database cluster and peptide entries with the shortest Euclidian distance. We demonstrate that the amino acid composition of peptides is virtually as informative as the sequence and allows rapid peptide identification more accurately than peptide mass alone. PMID- 15359737 TI - Analyzing alkaline proteins in human colon crypt proteome. AB - Normal human colon crypt protein extract was resolved by two-dimensional gel electrophoresis using pH 6-11 immobilized pH gradient strips in the first dimension. The optimized isoelectric focusing protocol includes cup-loading sample application at the anode and 1.2% hydroxyethyl disulfide (DeStreak), 15% 2 propanol and 5% glycerol in the rehydration buffer. Spots were well resolved across the entire pH range up to 11. A total of 311 protein spots were identified by mass spectrometry and peptide mass mapping. After combining isoforms, 231 nonredundant proteins were grouped into 16 categories according to their subcellular locations, and 17 categories according to their physiological functions. Histone proteins, ribosomal proteins and mitochondrial proteins were among the well-resolved highest p/ proteins. Application of this protocol to the analysis of normal and neoplastic colon crypts will contribute to the proteomic study of colorectal tumorigenesis since a significant portion of the human proteins is in basic pH range. PMID- 15359738 TI - Using immobilized metal affinity chromatography, two-dimensional electrophoresis and mass spectrometry to identify hepatocellular proteins with copper-binding ability. AB - To further our knowledge of intracellular copper transport, we used a proteomics strategy to search for hepatic proteins with copper-binding ability. Hep G2 cytosolic and microsomal fractions were applied to a copper(II)-loaded immobilized metal-affinity chromatography (IMAC) column. Protein identification was performed with 2-D gel electrophoresis and mass spectrometry. We identified 48 cytosolic proteins and 19 microsomal proteins displaying copper-binding ability. These proteins are diverse in function. Fifty-two of the 67 proteins contain putative metal-binding domains. We have identified many components of the Hep G2 copper metalloproteome including a large number of proteins not previously known to bind copper. PMID- 15359739 TI - Lectin affinity as an approach to the proteomic analysis of membrane glycoproteins. AB - The aim was to determine the proportion of membrane glycoproteins captured using concanavalin A or wheat germ agglutinin lectin affinity chromatography. Digests of the isolated proteins were separated by reversed-phase liquid chromatography and analyzed by matrix-assisted laser desorption tandem mass spectrometry. The two lectins identified different groups of proteins with a broad range of molecular mass and p/ values, including a number of proteins that overlapped the two groups. Approximately 30% of the proteins were positively identified as containing domains that were predicted using standard bioinformatics methods to be characteristic of integral membrane proteins. This approach represents an effective method of surveying the membrane protein pool of mammalian cells for subsequent proteomic analysis. PMID- 15359740 TI - Quantitative mass spectrometric immunoassay of insulin like growth factor 1. AB - Reported in this work are the development of mass spectrometric immunoassay (MSIA) devices and methods for the qualitative analysis of IGF-1 and -2, and the rigorous quantification of IGF-1 from human plasma. A method involving addition of SDS in moderate concentration to unfractionated plasma for disrupting IGF/IGFBP complexes was initially developed. The method is suitable for the direct extraction of the IGFs and subsequent mass spectrometric analysis. Rat plasma, containing IGF-1 that is mass shifted from human IGF-1, was used as an internal reference standard (IRS) for the quantification of IGF-1 directly from human plasma. A standard curve with linear dynamic range of at least 2 orders of magnitude was constructed from serially diluted IGF-1 standards containing equal amounts of rat plasma. Using the standard curve, IGF-1 levels in plasma samples from eight individuals were determined. The limit of detection for the IGF-1 MSIA was also evaluated and established to be approximately 15 pM. The assay is rapid and can be performed in parallel via high-throughput robotics processing. Furthermore, the mass spectrometry aspect of the developed IGF-1 immunoassay offers a new dimension in the ongoing study of IGF-1 and related diseases. PMID- 15359741 TI - Insights from modeling three-dimensional structures of the human potassium and sodium channels. AB - Ion channels allow the movement of ions across cell membranes. Nearly all cells have membranes spanned by ion channels, without which human nerves simply would not work. Ion channels are formed by the aggregation of subunits into a cylindrical configuration that allows a pore, thus forming a kind of tube for ion trafficking. In the present study, the subunits of the human potassium channel are formed by four identical protein chains, whereas for the case of the human sodium channel, the corresponding subunits are actually four hetero-domains formed by the folding of a very large but single protein chain. Since both of the two ion channels are important targets for drug discovery, the 3D (dimensional) structures of their pore regions were developed. On the basis of the 3D models, some important molecular biological mechanisms were discussed that may stimulate novel strategies for therapeutic treatment of the diseases related to ion channel disorders, such as long QT syndrome and chronic pain. PMID- 15359742 TI - Analysis of murine natural killer cell microsomal proteins using two-dimensional liquid chromatography coupled to tandem electrospray ionization mass spectrometry. AB - This study describes the application of a single tube sample preparation technique coupled with multidimensional fractionation for the analysis of a complex membrane protein sample from murine natural killer (NK) cells. A solution based method that facilitates the solubilization and tryptic digestion of integral membrane proteins is conjoined with strong cation exchange (SCX) liquid chromatography (LC) fractionation followed by microcapillary reversed-phase (microRP) LC tandem mass spectrometric analysis of each SCXLC fraction in second dimension. Sonication in buffered methanol solution was employed to solubilize, and tryptically digest murine NK cell microsomal proteins, allowing for the large scale identification of integral membrane proteins, including the mapping of the membrane-spanning peptides. Bioinformatic analysis of the acquired tandem mass spectra versus the murine genome database resulted in 11,967 matching tryptic peptide sequences, corresponding to 5782 unique peptide identifications. These peptides resulted in identification of 2563 proteins of which 876 (34%) are classified as membrane proteins. PMID- 15359743 TI - Self-contained on-chip cell culture and pretreatment system. AB - In this study, we describe a simple on-chip cell culture and pretreatment system that requires no external machines. Conventional cell culture utilizes culture dishes or microtiter plates, where pipetting and centrifugation are indispensable for washing cells and changing media. However, our microdevice requires no external centrifugation or pump. Utilizing this microdevice, we attained dramatically shorter total analytical time with a high-throughput screening system for proteomic analysis (1 min per 12 samples; one eightieth of the conventional time). Protein expression of Jurkat cells during stress-shock induced apoptosis was readily analyzed using this system. We found that a seaweed extraction effectively induced apoptosis of Jurkat cells. PMID- 15359744 TI - Analysis of altered protein expression patterns of Chlamydia pneumoniae by an integrated proteome-works system. AB - We have identified, analyzed, and quantified differential protein expression profile of five C. pneumoniae proteins, Adk (adenylate kinase), AhpC (thiol specific antioxidant), CrpA (15 KD cysteine rich protein), Map (methionine aminopeptidae), and Cpn0710 (hypothetical protein) under normal versus persistent growth conditions induced by interferon-gamma, at different time intervals of their replicative cycle by successfully employing the latest proteomic analysis tool, PDQuest 2-D analysis software. We have also determined that this software represents a reliable analytical tool for mapping protein expression patterns in C. pneumoniae. PMID- 15359745 TI - Explorative study of the protein composition of amniotic fluid by liquid chromatography electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. PMID- 15359746 TI - Proteomic analysis of human leukemic U937 cells incubated with conditioned medium of mononuclear cells stimulated by proteins from dietary mushroom of Agrocybe aegerita. PMID- 15359747 TI - DICOM modality worklist: an essential component in a PACS environment. AB - The development and acceptance of the digital communication in medicine (DICOM) standard has become a basic requirement for the implementation of electronic imaging in radiology. DICOM is now evolving to provide a standard for electronic communication between radiology and other parts of the hospital enterprise. In a completely integrated filmless radiology department, there are 3 core computer systems, the picture archiving and communication system (PACS), the hospital or radiology information system (HIS, RIS), and the acquisition modality. Ideally, each would have bidirectional communication with the other 2 systems. At a minimum, a PACS must be able to receive and acknowledge receipt of image and demographic data from the modalities. Similarly, the modalities must be able to send images and demographic data to the PACS. Now that basic DICOM communication protocols for query or retrieval, storage, and print classes have become established through both conformance statements and intervendor testing, there has been an increase in interest in enhancing the functionality of communication between the 3 computers. Historically, demographic data passed to the PACS have been generated manually at the modality despite the existence of the same data on the HIS or RIS. In more current sophisticated implementations, acquisition modalities are able to receive patient and study-related data from the HIS or RIS. DICOM Modality Worklist is the missing electronic link that transfers this critical information between the acquisition modalities and the HIS or RIS. This report describes the concepts, issues, and impact of DICOM Modality Worklist implementation in a PACS environment. PMID- 15359748 TI - A comparison of two methods of analog-to-digital medical video conversion. AB - The purpose of this study was to compare 2 methods of analog-to-digital video conversion in anticipation of improving, refining, and standardizing digital video production for medical education, diagnosis support, and telemedicine. A video workstation was devised containing 2 analog-to-digital video conversion systems: a digital video media converter with fire wire card and a video capture card. A procedure for final digital video production was created that used equivalent compression, pixel resolution, frame rate, and data rate for both systems. A subjective test was performed in which 12 archived analog videotapes, consisting of magnetic resonance angiograms, ultrasounds, neurosurgeries, and telemedicine applications, were converted digitally using the 2 methods. Randomized side by side video comparisons were analyzed and rated by subjective quality. An objective test was performed by videotaping a gray-scale test pattern off a computer monitor, digitally converting it by the 2 methods, and comparing the gray-scale values to the original pattern. There was no significant difference in overall video quality (P = .31) or grayscale reproduction using the 2 methods of analog-to-digital conversion. When performing simple analog-to digital video conversion, a video capture card is equal in quality and costs less than a digital video (DV) media converter or fire wire card. If a digital video camera is available for use, then a DV media converter or fire wire card is more advantageous because it enables full operation of the digital video camera. PMID- 15359749 TI - Frequency and impact of high-resolution monitor failure in a filmless imaging department. AB - The purpose of this study was to assess the image quality and the rate of failure of the high-resolution (2,048 x 1536 pixel) monitors used for primary diagnosis in a filmless radiology department and to analyze the type of problems encountered as well as the action taken to repair the monitors. Data were collected from Picture Archival and Communication System (PACS) service logs to determine rates of monitor adjustment and replacement, the symptoms reported, and the action taken. Additionally, random surveys of the high-resolution monitors were performed using a standard test pattern to assess spatial and contrast resolution in the center and outer corners of the monitors. Analysis of monitor service records showed a high rate of monitor replacement (41% per year) resulting in a relatively short "life expectancy" (defined as average time required before replacement) of 2.4 years. Random surveys of monitor quality using a standard test pattern showed suboptimal image quality in approximately 54% of the monitors with moderate image quality degradation present in at least one region of 27% of the high-resolution monitors, despite our vendor's quality control program. The results of this study support our subjective impression and those of other colleagues in the PACS community of an unacceptably high monitor failure rate and persistent image quality problems with 2,000 pixel monitors used for primary diagnosis. The relatively high incidence of suboptimal quality monitors suggests that more frequent quality control should be performed using a test pattern particularly given the fact that radiologists often are unable to discern degradation of monitor performance using clinical images. The high incidence of problems with image quality on high-resolution monitors indicates that vendors need to develop better quality control in monitor design and testing. Radiologists should review briefly a test pattern on each monitor at the beginning of each day. A computer program should be incorporated into the PACS, which asks radiologists to evaluate a test pattern and records the results in a central database, which is communicated to the service engineers. Further studies should be evaluated to determine the clinical impact of monitor image degradation, which is relatively easily seen using a test pattern but may be difficult to discern on clinical images. Requests for proposals (RFPs) for PACS and service contracts must specify carefully requirements for monitor image quality and conditions under which the vendor is required to replace these monitors. PMID- 15359750 TI - Network latency and operator performance in teleradiology applications. AB - Teleradiology applications often use an interactive conferencing mode with remote control mouse pointers. When a telephone is used for voice communication, latencies of the data network can create a temporal discrepancy between the position of the mouse pointer and the verbal communication. To assess the effects of this dissociation, we examined the performance of 5 test persons carrying out simple teleradiology tasks under varying simulated network conditions. When the network latency exceeded 400 milliseconds, the performance of the test persons dropped, and an increasing number of errors were made. This effect was the same for constant latencies, which can occur on the network path, and for variable delays caused by the Nagle algorithm, an internal buffering scheme used by the TCP/IP protocol. Because the Nagle algorithm used in typical TCP/IP implementations causes a latency of about 300 milliseconds even before a data packet is sent, any additional latency in the network of 100 milliseconds or more will result in a decreased operator performance in teleradiology applications. These conditions frequently occur on the public Internet or on overseas connections. For optimal performance, the authors recommend bypassing the Nagle algorithm in teleradiology applications. PMID- 15359751 TI - Experience with implementation of a radiology speech recognition system. AB - Recent advances in speech recognition technology have allowed development of computer systems for real-time radiologist-driven generation of reports. The transition to a speech recognition system is a technically complex process with many potential pitfalls that can decrease efficiency and disrupt workflow. In our recent experience with installation of such a system in an academic radiology department, factors that have worked against optimal performance have included environmental logistics, hardware incompatibilities, radiology information system interface problems, lack of suitable training, and inadequate technical support. Communication of our experience is intended to allow radiologists to anticipate complications of these systems and make informed decisions regarding the feasibility of such a system in their practices. With this information, potential buyers should be able to carefully scrutinize specifications for prospective systems and, by avoiding many of the possible pitfalls, make an easier transition to a speech recognition environment. PMID- 15359752 TI - Establishing benchmarks for creation of a pro-forma economic model to evaluate filmless PACS operation. AB - The purpose of this study was to establish data points (benchmarks) to incorporate into a pro-forma cost analysis model, comparing film-based and filmless modes of operation. Prospective data were collected over a 6-year period at the Baltimore VA Medical Center (BVAMC) immediately before and after implementation of a hospital-wide PACS. These data were in turn compared with local and national VA centers during comparable time periods, to establish reference data between manual film-based (without PACS) and filmless operations (using PACS). Benchmarks utilized for the study fell into 2 broad categories: operational costs and revenues generated. Factors contributing to operational costs include space requirements, equipment, supplies, personnel, and maintenance. Factors contributing to revenues generated included examination volume, modality mix, and reimbursement rates. Collectively, these data points were incorporated into a pro-forma model that allows prospective PACS customers to compare total cost of ownership for film-based and filmless operations dependent on the unique variables of the respective institution. PMID- 15359753 TI - Coping with PACS downtime in digital radiology. AB - As radiology departments become increasingly reliant on picture archiving and communication systems, they become more vulnerable to computer downtime that can paralyze a smoothly running department. The experiences and strategies developed during various types of picture archiving and communication system (PACS) downtime in a large radiology department that has completely converted to soft copy interpretation in all modalities except mammography are presented. Because these failures can be minimized but not eliminated, careful planning is necessary to minimize their impact. PMID- 15359754 TI - [Present status of therapy for viral hepatitis in Japan]. PMID- 15359755 TI - [Studies leading to the discovery of hepatitis C virus]. PMID- 15359756 TI - [Ultrastructural features of the HCV particle and HCV-like particles]. PMID- 15359757 TI - [Crystal structure of hepatitis C virus NS5B polymerase]. PMID- 15359758 TI - [Gene organization of hepatitis C virus and its sequence diversity]. PMID- 15359759 TI - [Genomic structure and function of untranslated region, structural region and non structural region of hepatitis C virus RNA]. PMID- 15359760 TI - [Structure and function of the internal ribosome entry site of HCV]. PMID- 15359761 TI - [Structural and functional characterization of the hypervariable region in the HCV genome]. PMID- 15359762 TI - [Initiation of genetic translation in HCV polyprotein]. PMID- 15359763 TI - [Processing of HCV precursor polyprotein]. PMID- 15359764 TI - [Hepatitis C virus core proteins and envelope (E1, E2) proteins]. PMID- 15359765 TI - [Structure and functions of HCV non-structural proteins]. PMID- 15359766 TI - [Genotyping of hepatitis C virus based on the full genome sequence]. PMID- 15359767 TI - [Molecular evolutionary analysis of hepatitis C virus--detection of positive selective sites]. PMID- 15359768 TI - [Mechanism of HCV-RNA replication]. PMID- 15359769 TI - [Translation stimulation and shut-off by HCV and viral multiplication]. PMID- 15359770 TI - [Role of interferon-sensitivity-determining region in replication of hepatitis C virus]. PMID- 15359771 TI - [Correlation of HCV-genome replication and mutation]. PMID- 15359772 TI - [Mode of proliferation of HCV in human lymphocytes]. PMID- 15359773 TI - [HCV culture systems in hepatocytes]. PMID- 15359774 TI - [Development of HCV infection system and HCV-RNA replication system]. PMID- 15359775 TI - [Establishment of cell lines expressing HCV replicon and its applications]. PMID- 15359776 TI - [HCV-N replicon and genome-length HCV-RNA replication system]. PMID- 15359777 TI - [Animal model for HCV infection--chimpanzee and tupaia, etc]. PMID- 15359778 TI - [Molecular dissection of hepatocarcinogenesis in HCV infection using transgenic animal models]. PMID- 15359779 TI - [Analysis of HCV core protein function using HCV core transgenic mice]. PMID- 15359780 TI - [Development of HCV vaccine]. PMID- 15359781 TI - [Mechanisms of the immune surveillance against HCV infection]. PMID- 15359782 TI - [Correlation between the grade of immune response against hepatitis C virus (HCV) infection and MHC class II polymorphisms]. PMID- 15359783 TI - [Mechanism of persistent infection of hepatitis C virus in humoral immune system]. PMID- 15359784 TI - [HCV-specific cytotoxic T lymphocyte and its epitope]. PMID- 15359785 TI - [Role of CTL in liver injury of patients with HCV infection]. PMID- 15359786 TI - [Relationship between the dynamics of cytokines and the phenotypes of the disease in chronic HCV infection]. PMID- 15359787 TI - [Involvement of dendritic cell dysfunction in the persistence of hepatitis C virus infection]. PMID- 15359788 TI - [Th1/Th2 balance in HCV-related liver cirrhosis and the effect of TGF-beta on Th1 response: possible implications for the development of hepatoma]. PMID- 15359789 TI - [Apoptosis and mechanisms in hepatitis C]. PMID- 15359790 TI - [Mechanisms and significance of apoptosis suppression by HCV core protein]. PMID- 15359791 TI - [Candidate molecules for HCV receptor]. PMID- 15359792 TI - [Mechanism of HCV cell entry mediated by envelope and receptor proteins]. PMID- 15359793 TI - [Characteristics of each commercially available anti-HCV assay kit used in screening for HCV infection]. PMID- 15359794 TI - [HCV core antigen assay and its clinical utility]. PMID- 15359796 TI - [Quantitative assay of HCV-RNA titers]. PMID- 15359795 TI - [Detection of hepatitis C virus RNA]. PMID- 15359797 TI - [In situ detection of hepatitis C virus: its clinical application]. PMID- 15359798 TI - [Comparison between HCV grouping by ELISA and genotyping by PCR method]. PMID- 15359799 TI - [Mutation of HCV-RNA and determination of its genome by RT-PCR]. PMID- 15359800 TI - [Epidemiology of hepatitis C in Japan]. PMID- 15359801 TI - [Significance of medical examination for HCV infection as a national project for prevention of hepatocellular carcinoma in Japan]. PMID- 15359802 TI - [Area and age specific prevalence rate of HCV infection in Japan]. PMID- 15359803 TI - [Routes of transmission of hepatitis C virus in hyperendemic areas, and molecular epidemiologic study of hepatitis C virus infection]. PMID- 15359804 TI - [Changes in the prevalence of HCV infection during 10 years in an HCV epidemic area of Japan]. PMID- 15359805 TI - [Present status of transfusion-associated hepatitis C virus infection--efficiency after implementation of nucleic acid amplification testing]. PMID- 15359806 TI - [Administrative policy concerning the issue of hepatitis virus infection by blood products]. PMID- 15359807 TI - [Natural course of hepatitis C in infants infected by their mothers]. PMID- 15359808 TI - [Mechanism and prevention of mother-to-infant transmission of HCV]. PMID- 15359809 TI - [Epidemiology of intrafamilial transmission of hepatitis C virus by molecular evolutionary analysis]. PMID- 15359810 TI - [Prevention of infection with hepatitis C virus in medical personnel as an aspect of risk management]. PMID- 15359811 TI - [Efficacy of short-duration interferon administration to prevent HCV transmission to medical personnel]. PMID- 15359812 TI - [Prevalence of hepatitis C virus infection among chronic hemodialysis patients]. PMID- 15359813 TI - [Effect of HCV infection in Japanese hemophiliacs--including influence of HTV-HCV coinfection on the prognosis of hemophiliacs]. PMID- 15359814 TI - [HCV infection among narcotics/methamphetamine abusers]. PMID- 15359815 TI - [Transmission of hepatitis C virus (HCV) by tattooing and acupuncture]. PMID- 15359816 TI - [Hepatitis C virus infection as a sexually transmitted disease]. PMID- 15359817 TI - [Demonstration of hepatitis C virus genome in saliva, urine and feces of patients with type C hepatitis]. PMID- 15359818 TI - [Guidelines for treatment of hepatitis C]. PMID- 15359819 TI - [Clinical view of acute hepatitis C]. PMID- 15359820 TI - [Early diagnosis of acute hepatitis C--detection of anti-HCV antibody and HCV RNA]. PMID- 15359821 TI - [Treatment of acute hepatitis C]. PMID- 15359822 TI - [Role of hepatitis C virus in fulminant hepatitis]. PMID- 15359823 TI - [Fulminant hepatitis C and subacute hepatitis C: pathological and clinical features]. PMID- 15359824 TI - [Treatment of fulminant hepatitis C or subacute hepatitis C]. PMID- 15359825 TI - [Clinical, virologic and pathologic features of asymptomatic hepatitis C virus carriers]. PMID- 15359826 TI - [Impact of HLA polymorphism on asymptomatic HCV carriers]. PMID- 15359827 TI - [Genetic variability and clinical significance of hypervariable region of hepatitis C virus]. PMID- 15359828 TI - [Differential diagnosis between asymptomatic HCV carriers and previously infected patients with anti-HCV antibody in general clinic setting]. PMID- 15359829 TI - [Clinical management of asymptomatic HCV carriers]. PMID- 15359830 TI - [General remarks on chronic hepatitis C]. PMID- 15359831 TI - [Chronic hepatitis C: virologic and immunologic aspects]. PMID- 15359832 TI - [Pathology of chronic hepatitis C and its histological features]. PMID- 15359833 TI - [Variation in quasispecies population of blood HCV-RNA in each stage of type C chronic liver disease]. PMID- 15359834 TI - [Measurement of strand-specific hepatic HCV-RNA quantities of patients with hepatitis C virus infection]. PMID- 15359835 TI - [Molecular analysis of hepatitis C virus in pathogenesis of chronic hepatitis C]. PMID- 15359836 TI - [Quantification of hepatitis C virus RNA in serum]. PMID- 15359837 TI - [Analysis of factors related to prognosis in patients with chronic hepatitis type C--clinical laboratory data]. PMID- 15359838 TI - [Histological findings in the liver in patients with chronic hepatitis C]. PMID- 15359839 TI - [Therapeutic strategies for hepatitis C]. PMID- 15359840 TI - [Human interferon: molecular mechanism of its antiviral activities]. PMID- 15359841 TI - [Antiviral mechanism of ribavirin against RNA virus]. PMID- 15359842 TI - [Multicenter study on the inhibition of hepatocarcinogenesis by interferon therapy (IHIT)]. PMID- 15359843 TI - [Combination therapy of interferon plus ribavirin for chronic hepatitis C]. PMID- 15359844 TI - [Virus factors determining the outcome of IFN treatment for chronic hepatitis C]. PMID- 15359845 TI - [Host factors determining the outcome of IFN treatment for chronic hepatitis C]. PMID- 15359846 TI - [Differential effects of interferon-based treatments in chronic hepatitis C]. PMID- 15359847 TI - [Evaluating the efficacy of interferon therapy on hepatitis C patients]. PMID- 15359848 TI - [Therapeutic effects of interferon on chronic hepatitis C from the aspect of dosage and method of administration]. PMID- 15359849 TI - [Two-step interferon rebound therapy]. PMID- 15359850 TI - [Efficacy of and indications for IFN retreatment of patients with chronic hepatitis C]. PMID- 15359851 TI - [Clinical significance of measuring HCV-RNA levels in patients with chronic hepatitis C by IFN treatment, including combination therapy with ribavirin]. PMID- 15359852 TI - [Cost-effectiveness of antiviral therapy for chronic hepatitis C patients in prevention of hepatocellular cancer ]. PMID- 15359853 TI - [Effectiveness of long-acting interferon (peginterferon)]. PMID- 15359854 TI - [Complications in interferon therapy for chronic hepatitis C]. PMID- 15359855 TI - [Prevention of adverse effects in ribavirin and interferon combination therapy]. PMID- 15359857 TI - [Phlebotomy, dietary iron restriction therapy and bovine lactoferrin therapy for chronic hepatitis C]. PMID- 15359856 TI - [Future antiviral therapy for chronic hepatitis C]. PMID- 15359858 TI - [Alternative measures for patients with chronic hepatitis C unresponsive to interferon therapy]. PMID- 15359859 TI - [Molecular mechanisms of extrahepatic lesions]. PMID- 15359860 TI - [Hepatitis C virus-associated glomerulonephritis]. PMID- 15359861 TI - [Pathogenesis of and therapy for extrahepatic lesions: myocarditis and cardiomyopathy]. PMID- 15359862 TI - [HCV and extrahepatic manifestations]. PMID- 15359863 TI - [Effect of IFN treatment for extrahepatic manifestations]. PMID- 15359864 TI - [Incidence and clinical statistics in HCV antibody-positive liver cirrhosis]. PMID- 15359865 TI - [Fibrosis progression in hepatitis C]. PMID- 15359866 TI - [Clinical aspects of type-C cirrhosis of liver--diagnosis, clinical course, treatment and prognosis]. PMID- 15359867 TI - [Progression and risk factors for hepatocarcinogenesis in hepatitis C-related cirrhosis--viral factors, host factors, and others]. PMID- 15359868 TI - [Effects of IFN therapy on incidence of hepatocellular carcinoma in HCV positive cirrhosis]. PMID- 15359869 TI - [Liver transplantation for hepatitis C virus cirrhosis--present status and future prospects]. PMID- 15359870 TI - [Epidemiology of hepatitis C virus and hepatocellular carcinoma in Japan]. PMID- 15359871 TI - [Clinical characteristics of hepatocellular carcinoma in patients with antibody to hepatitis C virus]. PMID- 15359872 TI - [Oncogenes and suppressor oncogenes in C-type hepatocellular carcinoma]. PMID- 15359873 TI - [Analysis of replicating hepatitis C virus quasispecies in hepatocellular carcinoma tissues]. PMID- 15359874 TI - [Recurrence of hepatocellular carcinoma associated with hepatitis C virus after tumor ablation]. PMID- 15359875 TI - [Hepatitis C in childhood--natural history and therapy]. PMID- 15359876 TI - [Interferon treatment for aged patients with hepatitis C]. PMID- 15359877 TI - [Chronic hepatitis C in hemodialysis patients]. PMID- 15359878 TI - [Diagnosis and treatment of alcoholic liver disease with HCV]. PMID- 15359879 TI - [Autoimmune hepatitis and HCV]. PMID- 15359880 TI - [Difference between autoimmune hepatitis (AIH) with and without hepatitis C virus (HCV)]. PMID- 15359881 TI - [Selection of therapy in patients with chronic hepatitis C showing features like autoimmune hepatitis]. PMID- 15359882 TI - [Prevalence and clinical course in HBV and HCV dual chronic infection]. PMID- 15359883 TI - [Progression of liver disease in HCV co-infected HBV-DNA carrier]. PMID- 15359884 TI - [Epidemiological study of HGV coinfection in patients with hepatitis C]. PMID- 15359885 TI - [Hepatitis C in the HIV-infected person]. PMID- 15359886 TI - [Physicochemical factors influencing distribution of Legionella species in Japanese hot springs]. AB - We examined the relationship between the distribution of Legionella bacteria and various physicochemical characteristics of hot springs in Japan. Legionella bacteria were isolated from 52 (49.5%) out of 105 water samples, particularly from outdoor hot springs (67.3%). The bacterial count in the water samples positive for Legionella (86.5%) ranged from 10(1) to < 10(3) cfu/100 mL. L. pneumophila serogroup (SG) 4 (27.8%) was predominant in the water samples, followed by SG 5 (12.2%). The pulsefield gel electrophoresis (PFGE) patterns of chromosomal DNA for L. pneumophila SG 4 isolated from different parts of a hot spring resort were identical. Isolation of Legionella species from hot spring waters did not occur at pH 1.8-3.3, SO4(2-): > 780 mg/L, and H2SiO3: > 146 mg/L. The hot water-recirculating systems were applied to 18 out of 20 (90%) hot spring facilities which were found positive for Legionella. These results indicate that Legionella species are widespread in hot springs throughout Japan, except for waters with a low pH and non-recirculating waters, and that a single strain of L. pneumophila SG 4 is predominant in a particular hot spring resort. PMID- 15359887 TI - [Survey of hepatitis B and C in students of faculty of dentistry and dental hygienist school]. AB - At present, in Japan, hepatitis B virus (HBV) and hepatitis C virus (HCV) infection by blood transfusion rarely happens. However, according to the national survey, outbreak of sporadic acute hepatitis B and C is reported every year and viral hepatitis induced by iatrogenic infection is also reported. We think that education and enlightenmen for measures of infection control for hepatitis virus in dentisal medical care are important. Therefore, we carried out a questionnaire survey about measures of an infection control including hepatitis B and C for 352 students of a certain faculty of dentistry and a dental hygienist school. 35.5% of the total students thought the defense of oneself against infection was more important than defense of cross infection. Furthermore, the prevalence of the student who thought to permit recycle of a disposer glove and a disposer cartridge of a local anesthesia was 13.1% (46/352), 14.8% (52/352), respectively. The prevalence of students who recognized that HCV and HBV were detected from not only blood but also body fluid such as saliva remained in 65.3%. Consequently, the reality that knowledge of hepatitis virus and understanding about sterilization and disinfection of instruments were low became clear. In conclusion, immediate making of the guideline that aimed at standardization of prevention of hospital infection in domestic dental treatment and education to introduce the curriculum with a high regard for risk management of infection for students of dentistry will be required. In addition, it is an important problem to spread thoughts of standard precautions for dentistry. PMID- 15359888 TI - [The performance of (1, 3)-beta-D-glucan and Aspergillus galactomannan measurement for early diagnosis of invasive aspergillosis in patients with hematological diseases]. AB - We analyzed the performance of (1, 3)-Beta-D-glucan (measurement by the alkaline kinetic chromogenic Limulus method (FUNGITEC G test-MK, Fungitec) and the kinetic turbidimetric Limulus method [Beta-Glucan test WAKO, Wako]) and we carried out Aspergillus galactomannan antigen detection (enzyme-linked immunosorbent assay, ELISA test) for the early diagnosis of invasive aspergillosis in patients with hematological diseases at the time of febrile episodes of unknown origin that did not respond to antibacterial therapy for more than 3 days. During a one-year period (April 2002 to March 2003), a total of 69 febrile episodes in 58 patients were studied; 8 cases of invasive aspergillosis were diagnosed according to the definition of the European Organization for Research and Treatment of Cancer/Mycosis Study Group, and 61 cases were found to be non-mycotic diseases. Based on the analysis of 69 results with confirmed disease status, the overall performance of the Fungitec, the Wako, and the ELISA test were as follows: sensitivity was 0.88, 0.63, and 0.50, respectively, whereas the specificity was 0.85, 0.98, and 1.0, respectively. Moreover, there was a strong relationship between the log-transformed values of the (1, 3)-Beta-D-glucan levels measured by the two methods (r = 0.92 [95%CI, 0.89-0.94] ; p<0.001). For the statistical analysis of these serological tests a receiver operating characteristic curve (ROC) was used, as well as the resulting area under the ROC curve (ROC AUC). The ROC AUC and the cut-off values that gave the highest accuracy were as follows: 0.92, 24.9 pg/ml for the Fungitec, 0.84, 7.3 pg/ml for the Wako, and 0.89, 0.9 COI for the ELISA test, respectively. In conclusion, these results indicate that both of the two (1, 3)-Beta-D-glucan measurement approaches served equally well as surveillance tools for determining the extent of invasive aspergillosis; in addition, the log-transformed value of these tests can be used for comparison. Moreover, the ELISA test was found to have clinical utility, both as a surveillance and as a diagnostic tool when invasive aspergillosis was suspected. It should be noted that the galactomannan assay had sensitivity-related limitations; lowering the cut-off value is expected to increase the diagnostic value for use in cases of invasive aspergillosis. PMID- 15359889 TI - [Evaluation of isolation media for the detection of Bartonella henselae- isolation of Bartonella henselae from domestic cats]. AB - Bartonella henselae is a causative agent of cat scratch disease. We preliminarily tested four media for the bacterial growth, including agar plates with sheep, horse or rabbit blood, and chocolate agar. Of these media, rabbit blood and chocolate agar plate were found to be more excellent for the growth than the medium with sheep or horse blood. Blood samples from 60 domestic cats in Yamaguchi Prefecture were then cultured using 7% rabbit blood agar plates and BACTEC9050 (BD), automated blood culture microbial detection system. B. henselae was isolated from six of the 60 (10%) blood samples. Tiny colonies of B. henselae were visible on the agar medium after one week of culture at 35 degrees C in the 5% CO2 atmosphere. BACTEC 9050 detected B. henselae in one of the 10 blood samples and it took two weeks to detect the bacteria automatically, though gram stain failed to show organisms in the blood culture bottle. In conclusion, rabbit blood or chocolate agar and incubation of agar media more than one week and of BACTEC more than two weeks are recommended for the detection of B. henselae. PMID- 15359890 TI - [Molecular epidemiologic analysis of nosocomocal infection due to TEIC resistant MRSA strains]. AB - We performed epidemiologic study of 109 strains of methicillin resistant Staphylococcus aureus (MRSA) which were detected in our hospital. Of these strains, 6 strains showed resistant to Teicoplanin (TEIC) which MIC level were between 4 to 8microg/mL. All of them showed some phenotype, such as type II of coagulase, type A of enterotoxin, and were producing TSST-1. Genotype analysis by PFGE also showed that those strains ware identical. From analyzing the spreading rout of these TEIC resistant MRSA, we speculate that they first were in ICU ward, then spread all over the hospital carried by the stuff cross-working ICU and other units of hospital. PMID- 15359891 TI - [The effects of fluorescent lamp coated with a photocatalyst film on the number of floating microorganisms or smell molecules in a room installed with the lamp]. AB - Fluorescent lamp that is coated with a photocatalyst film, titanium dioxide (TiO2), can catalyze the microorganisms and smell producing molecules that touch the membrane. In this report, we examined the effect of this coated lamp by the number of floating microorganisms or smell molecules in a room installed with this lamp. The number of floating microorganisms was examined independently in different laboratories before and after installing the lamps in an animal facility and a vegetable processing room of a food company. We found that the number of floating microorganisms was significantly reduced after the installation of this coated lamp. To test for smell molecules, acetaldehyde was injected into a closed chamber installed with the coated lamp and the result was compared to a control where a regular fluorescent lamp was used. The acetaldehyde concentration did not change with the regular lamp but was halved after 1h with the photocatalyst coated lamp. To test the effect in real settings, questionnaires were filled out by restroom users in several different places. About 90% of those surveyed answered that there was a reduction of smell after the installation of the photocatalyst coated lamp. We concluded that the installation of this coated fluorescent lamp reduced not only the floating microorganisms but the smell in various practical situations. PMID- 15359892 TI - [Evaluation of an immunochromatography test using enzyme immunoassay for rapid detection of influenza A and B viruses]. AB - We evaluated the performance of an improved version of Espline Influenza A & B-N (Fujirebio Inc., Japan), an immunochromatography test using enzyme immunoassay for rapid diagnosis of influenza A and B. The test produced positive results for four strains of influenza viruses and thirty-one influenza viral antigens and negative results for all of thirty strains of other respiratory viruses that were tested. The detection limit of this test was 5.8 x 10(2) to 5.8 x 10(3) pfu/assay, which is more sensitive than the old version of Espline. Furthermore, 715 respiratory specimens collected from the patients (children, 79.4%; adults, 18.5%; unknown, 2.1%) with influenza-like illnesses during the 2002/2003 influenza season in Japan were tested as part of a clinical evaluation of this test. The relative performance of this test compared to cell culture and nested RT-PCR results were examined. In the cell cultures, influenza viruses were detected in 488 of the 715 specimens (overall, 68.3%; AH3, 41.7%; B, 26.4%; AH3 and B, 0.1%). For influenza A, the sensitivity of this test was 95.4% (125/131) for nasal aspirates, 96.8% (92/95) for nasal swabs, and 85.1% (63/74) for throat swabs. For influenza B, the sensitivity of this test was 91.2% (52/57) for nasal aspirates, 88.1% (59/67) for nasal swabs, and 71.6% (48/67) for throat swabs. The new test exhibited a remarkably higher sensitivity to influenza A in throat swabs than the old version of Espline. Only two false positive results were obtained out of a total of 223 virus negative specimens; the specificity of the test was 100% (88/88) for nasal aspirates, 97.6% (81/83) for nasal swabs, and 100% (52/52) for throat swabs. We conclude that the new Espline Influenza A&B-N rapid diagnostic test is easy to use and has a high sensitivity and specificity, especially for influenza A. PMID- 15359893 TI - [Successful treatment of beta-lactamase-negative ampicillin-resistant Haemophilus influenzae type b meningitis with high-dose ceftriaxone administration]. AB - Haemophilus influenzae type b (Hib) is the most frequent pathogen of bacterial meningitis in Japanese children. The prevalence of beta-lactamase-negative ampicillin-resistant (BLNAR) Hib strain has been increasing in recent years. Furthermore, antibiotic activities of cefotaxime (CTX) and ceftriaxone (CTRX) have decreased against some of those BLNAR strains. We report a case of one-year old boy who suffered from meningitis caused by BLNAR Hib. The MICs of CTX and CTRX for the strain isolated from cerebrospinal fluid was 1.0 and 0.5 microg/ml, respectively. The patient was administered high-dose CTRX (150 mg/kg/day) and recovered completely without any sequela. The high-dose CTRX administration may be a considerable choice of the treatment of BLNAR meningitis. PMID- 15359894 TI - [Researches on health and welfare promotion based on an approach of social medicine]. AB - This article was derived from my memorial talk given when receiving the prize of the Japanese Society for Hygiene at their academic congress. The reader could review my research on health and welfare promotion made by introducing new conceptual health policy based on the approach of social medicine. Through my experience in different research work, the importance of social factors in the etiology of health during childhood, adulthood and old age was discussed. In addition, it was revealed that social factors not only influence the population's health status but also constitute the context within which organized efforts can be made to promote health. For the elderly, the annual health check, stroke patient registration, and insurance for care and spousal bereavement; for adults, the Karoshi and occupational health; and for children, air pollution-atopy predisposition and lifestyles were highlighted as social medicine-related issues. The research on mostly longitudinal population studies showed that health status, including the life expectancy and the prevalence of disability and chronic disorders, are related to one's marital status, social support, psychosocial working conditions and environmental factors as well as to lifestyles such as physical activity and hours of work and sleep at entry. More attention should be directed to independent factors' effects on health, separate from those of adverse health habits and bio-medical situations, under the health and welfare promotion strategy. PMID- 15359895 TI - [Risk assessment of environmental carcinogenesis using WBC DNA adducts]. AB - Environmental carcinogens are enzymatically activated to form intermediates that can react with cellular DNA and form DNA adducts. Several kind of carcinogens bind to several sites of DNA. The measurement of WBC DNA adducts is a useful indicator for environmental carcinogen exposure monitoring. The 32P-Postlabeling method is a most popular and very sensitive method for DNA adduct analysis. We can analyze 1 adduct/10(8) nucleotides. In this review, I show some data of DNA adduct analysis for PAH exposure. These results demonstrated that inter individual variation was very large. There were some confounding factors, such as metabolism or repair variations. I also showed some limitations of DNA adduct analysis. The method of adduct analysis is very complicated with several steps. We need to improve the accuracy. Do the data from WBC explain the target organ, such as the lung or liver? Almost all previous studies have been cross-sectional. We need a large-sized cohort study to evaluate whether adducts are a predictor of cancer. DNA adducts should be an important factor in gene-environment interaction. PMID- 15359896 TI - [Toxicological significance of metallothionein on environmental harmful factors: verification and suggestions from a metallothionein-I/II null mouse model study]. AB - In many research groups including our laboratory, metallothionein (MT)-I/II null mice have been used to clarify the biological function and physiological role of MT. Recent studies with MT-I/II null mice concerning the role of MT in the toxicity and distribution of metal, oxidative stress and chemical carcinogenesis were reviewed. Some reports, including our findings, showed that MT-I/II null mice have an increased sensitivity to harmful metals such as cadmium, mercury, zinc and arsenic. Moreover, it was clarified using MT-I/II null mice that MT plays a major role in the retention of cadmium, mercury and zinc in target tissues. MT-I/II null mice were found to be much more sensitive than wild-type mice to the toxicity caused by free radical-inducing factors, which include paraquat, acetaminophen, ethanol, X-ray, ultraviolet B, carbon tetrachloride, cisplatin, doxorubicin, cerulein and streptozotocin. In addition, MT-I/II null mice were highly susceptible to skin carcinogenesis induced by 7,12 dimethylbenz[a]anthracene and bladder carcinogenesis caused by N-butyl-N-(4 hydroxybutyl)nitrosamine. These results suggest that MT is an important protective factor against metal toxicity, oxidative stress and chemical carcinogenesis. PMID- 15359897 TI - [Effects of gender-related weight reduction on the physical condition of male and female college judoists]. AB - OBJECTIVE: In order to study the gender-related effects of weight reduction on the physical condition of male and female college judoists during weight reduction periods, we examined changes in anthropometric, nutritional intake and biochemical parameters in 43 male and female college judoists 20 days (pre-weight reduction) before and one day (post-weight reduction) before the game. METHODS: Subjects were divided into two groups. Thirty subjects (22 males, 8 females) who required weight reduction were defined as the weight reduction (WR) group, and the remaining 13 subjects (5 males, 8 females) were defined as the non-reduction (non-WR) group. RESULTS: In both WR groups, anthropometric parameters and nutritional intake significantly decreased after weight reduction compared to the pre-values. The change ratios of nutritional intake during the weight reduction period in the female WR group were higher than those in the male WR group. Serum lipids and electrolytes significantly decreased after the weight reduction compared to the pre-values in both WR groups. In the female WR group, Cr and UA significantly increased after the weight reduction compared to the pre-values. Although it was not significant, Ht increased after the weight reduction compared to the pre-values only in the female WR group. TP, Cr, UA, BUN, CK, LDH and IgA significantly increased after the weight reduction compared to the pre-values in the weight reduction compared to the pre-values in the male WR group. In contrast, in both non-WR groups, these parameters did not change significantly during the research period. CONCLUSIONS: Our findings suggest that nutritional exhaustion and dehydration may be the major effects brought about by energy and fluid restriction in female college judoists during a weight reduction period. On the other hand, in male college judoists undergoing weight reduction, the data suggest that muscle damage and myogenic protein catabolism may be caused chiefly by exercise. These differences may be due to the difference in the method of weight reduction for the different genders, namely, mainly exercise in males and mainly energy and fluid restriction in females. PMID- 15359898 TI - [Pharmacoeconomic analysis of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor therapy for the secondary prevention of coronary heart disease]. AB - OBJECTIVE: The pharmacoeconomics of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (pravastatin) therapy were evaluated using data from an open labeled, randomized, controlled trial (PCS: Prevention of Coronary Sclerosis study) with a two-year follow-up period. METHODS: Patients with a serum total cholesterol concentration of between 180 and 219 mg/dL at baseline were randomly assigned to two groups: the pravastatin group (n=54) and the group that did not receive cholesterol-lowering drugs (no-pravastatin group, n=66). A cost minimization analysis and a cost-effectiveness analysis were then performed from the perspective of a health insurance company, using insurance information obtained from the subjects. In the cost-minimization analysis, the expected costs associated with percutaneous transluminal coronary angioplasty (PTCA) and the pravastatin prescription were compared; the total medication costs between the two groups were also compared. In the cost-effectiveness analysis, the estimated savings per event-free year were calculated. RESULTS: The median cost of a PTCA procedure was 1,323,492 yen. The expected insurance cost for two years was 182,532 yen per patient in the pravastatin group and 224,444 yen per patient in the no-pravastatin group. The cost-minimization analysis showed that the cost of healthcare for the no-pravastatin group was higher than that for the pravastatin group. The mean total cost of medication was 729,849 yen per patient in the pravastatin group and 989,606 yen per patient in the no-pravastatin group. The cost-effectiveness ratio for coronary heart disease was 2,766,994 yen. CONCLUSION: Pravastatin therapy in normocholesterolemic patients with coronary sclerosis seems to have an excellent pharmacoeconomic profile. PMID- 15359899 TI - [Regional differences in perinatal mortality rates in Japan--an investigation based on vital statistics]. AB - OBJECTIVE: This study investigated whether regional differences in perinatal mortality rates are related to distances to medical resources for perinatal care, such as the Neonatal Intensive Care Unit (NICU). METHODS: Using the census for Japan in 2000, all municipalities in Japan were classified into four groups based on population size: municipalities of 1,000,000 people or more (Group 1), municipalities between 350,000 and 1,000,000 people (Group 2), municipalities between 10,000 and 350,000 people (Group 3), and municipalities of less than 10,000 people (Group 4). Then, using the vital statistics, perinatal mortality rates for all groups were calculated. In addition, setting the perinatal mortality rate in Group 1 as a referent, we calculated the differences in the rates to detect regional differences in perinatal mortality in Japan. Finally, we compared the distances between the municipality center and the closest NICU among the four groups by one-way ANOVA. RESULTS: The perinatal mortality rates of Groups 1, 2, 3 and 4 were 5.38, 5.58, 5.88 and 6.31, respectively. The perinatal mortality rate ratios (95%CI) were 1.04 (0.96-1.12), 1.09 (1.03-1.16), and 1.17 (1.05-1.31), respectively. The perinatal mortality rate differences (95%CI) were 0.20 (-0.24-0.64), 0.50 (0.15-0.84), and 0.93 (0.23-1.62), respectively. The distances (95%CI) between the center of the municipalities and the closest NICU for Groups 1, 2, 3 and 4 were 2.61 (1.56-3.66) km, 4.23 (2.74-5.72) km, 20.79 (17.68-23.90) km and 38.07 (31.65-44.48) km, respectively. All of the differences among the groups were significant (p < 0.001), except for the difference between Group 1 and Group 2. CONCLUSION: There are regional differences in perinatal mortality rates in Japan. This study suggests that the difference is associated with the uneven distribution of NICUs. PMID- 15359901 TI - [The relationship between Staphylococcus aureus and atopic keratoconjunctivitis]. AB - PURPOSE: To investigate the role of Staphylococcus aureus in patients with vernal keratoconjunctivitis (VKC) and atopic keratoconjunctivitis (AKC) complicated by atopic dermatitis (AD). SUBJECTS AND METHODS: Microbiological culture from the conjunctival sac of acute exacerbated patients with VKC and AKC complicated by AD was performed. The subjects were 29 patients (31 eyes) with VKC and AKC who showed acute exacerbated clinical symptoms of conjunctivitis. Antigen specific IgE antibodies to staphylococcal enterotoxin A (SEA) and staphylococcal enterotoxin B (SEB) in serum were also examined. In this study, the patients were divided into two groups: An AD group consisting of 22 patients with VKC and AKC complicated by AD, and a control group consisting of 8 patients with allergic conjunctivitis without AD. We also examined SEA/SEB specific IgE antibody in tears from the 5 patients who underwent bacteriological examinations of the conjunctival sac at the same time. RESULTS: Twenty-seven out of 31 eyes were gram positive in the bacteriological culture from the conjunctival sac (87.1%). Staphylococcus aureus was detected in 21 out of the 27 eyes (77.8%). In the AD group, 2 of the 22 cases were gram-positive for serum SEA specific IgE antibodies, 2 cases of the 22 cases were gram-positive for SEB specific IgE antibodies, and 9 cases were gram-positive for both SEA and SEB specific IgE antibodies. Serum SEA and SEB specific IgE antibodies were all gram-negative in the control group. Either SEA or SEB specific IgE antibody in tears was detected in all of the above 5 patients who underwent bacteriological examinations of the conjunctival sac, and 4 of the 5 were gram-positive. Staphylococcus aureus was isolated in 3 out of the 5 patients, and 1 case was gram-negative. CONCLUSION: Staphylococcus is one of the exacerbating factors in VKC and AKC. It is important to evaluate both bacteriological examinations of the conjunctival sac and SEA/SEB specific IgE in tears. PMID- 15359900 TI - [New informations on medical treatment for normal-tension glaucoma]. PMID- 15359902 TI - [Comparison of the effects of latanoprost and timolol gel-forming solution on diurnal variation of intraocular pressure in normal-tension glaucoma]. AB - OBJECTIVE: To compare the effects of latanoprost (Lat) and timolol gel-forming solution (Tg) on the diurnal variation of intraocular pressure (IOP) in normal tension glaucoma (NTG). SUBJECTS AND METHODS: A total of 47 NTG patients (47 eyes) were randomly assigned to receive Lat alone (25 eyes) or Tg alone (22 eyes). IOP was measured at fixed time points during 24 hours before and after treatment with each drug. In addition, blood pressure and pulse rate were measured before and after treatment. RESULTS: Lat reduced IOP significantly at all time points of measurement. Tg reduced IOP significantly at all time points of measurement except at 22:00 and 03:00. Percent reductions in the IOP at 03:00 and in the mean diurnal IOP were significantly greater in the Lat group than in the Tg group. In the Lat group there was no change in diastolic pressure or pulse rate after treatment, but in the Tg group these parameters decreased significantly. CONCLUSIONS: Comparison of diurnal variation of IOP before and after treatment shows that Lat is more effective than Tg in lowering IOP. In addition, Lat does not affect blood pressure or pulse rate. PMID- 15359903 TI - [Effect of ocular hypotensive medications in preventing the progression of visual field damage in patients with normal-tension glaucoma]. AB - PURPOSE: We investigated the effect of ocular hypotensive medications and the risk factors for progression of visual field damage in patients with normal tension glaucoma (NTG). SUBJECTS AND METHODS: Sixty-four NTG patients (64 eyes) followed at least 4 years with ocular hypotensive medications were studied regarding the probability of visual field stability using the Kaplan-Meier life table analysis and multiple regression analysis with the Cox proportional hazards model. RESULTS: Probability of visual field stability was 56% at the 76-month follow-up. Mean intraocular pressure during the follow-up period and generalized cup enlargement type classified according to the optic disc shape were found to have significant influence on the progression of visual field damage. CONCLUSION: These findings suggest that ocular hypotensive medications are effective in stabilizing the visual field. We need to pay attention to the optic disc shape in patients with NTG. PMID- 15359904 TI - [Histopathological findings in cytomegalovirus retinitis]. AB - PURPOSE: We examined eyeballs collected from autopsied acquired immunodeficiency syndrome patients with cytomegalovirus (CMV) retinitis, and analyzed the precise pathogenesis of CMV retinitis. MATERIAL AND METHODS: Eyeballs were fixed with 10% buffered formalin embedded in paraffin. CMV antigens were investigated by histopathological and immunohistochemical analyses. Histopathological findings were compared with funduscopic images. RESULTS: CMV antigens remained in the necrotic area of the retina and many CMV immediate early antigens existed in intact parts of the inner retina showing almost intact structure, and around retinal vessels. CONCLUSIONS: The results suggest that CMV infects the inner retina first via the retinal vessels, although funduscopic examination may appear normal. It extends through the neuronal cells and glial cells horizontally and Muller cells vertically. CMV severely damages the retinal structure. PMID- 15359906 TI - [On: Multifocal electroretinograms in patients with retinal vein occlusion]. PMID- 15359905 TI - [Clinicopathological study on suprachoroidal and supraciliary hemorrhage during enucleation]. AB - PURPOSE: To investigate the pathogenesis of suprachoroidal and supraciliary hemorrhage that might have been induced during enucleation. METHODS: A histopathological examination of 392 enucleated eyeballs was carried out and 8 eyeballs with suprachoroidal and supraciliary hemorrhage were selected for further clinicopathological examination. RESULTS: Among 14 eyeballs with severe acute intraocular inflammation, 7 eyeballs with suprachoroidal and supraciliary hemorrhage were found and one other such eyeball was seen among 53 with neovascular glaucoma. Among these 8, there was one case of prolapse of intraocular tissue with severe hemorrhage into the suprachoroidal and supraciliary spaces; 4 cases of severe hemorrhage into the suprachoroidal and supraciliary spaces without prolapse of intraocular tissue; and 3 cases of mild hemorrhage into the suprachoroidal or supraciliary spaces. CONCLUSIONS: In the eyeballs with severe acute intraocular inflammation, intraocular pressure was elevated and the blood vessels were weakened by inflammatory cell infiltration. During enucleation external forces affected the blood vessel wall of the ciliary arteries and vortex veins, and the breakdown of vessel walls might have been the cause of the suprachoroidal and supraciliary hemorrhage. PMID- 15359907 TI - [A review 19 Hereditary retinal degeneration]. PMID- 15359909 TI - [Cell adhesion molecules in multiple myeloma]. PMID- 15359908 TI - [Application of WT1 transcript assay for allogeneic stem cell transplantation for acute leukemia]. PMID- 15359910 TI - [Role of polymorphic adhesion molecules in the development of graft-versus leukemia effect after HLA-matched allogeneic stem cell transplantation]. AB - Mismatches of minor histocompatibility antigens (mHas) between HLA-identical stem cell donors and recipients are known as a major risk factor for graft-versus-host disease (GVHD). We determined the alleles of 5 polymorphic molecules including HA 1 and 4 adhesion molecules in 102 patients who had undergone stem cell transplantation from HLA-identical donors and investigated the association of their mismatches with the relapse rate and incidence of GVHD. We observed relapse rates of 16.1% in patients with at least one or more incompatibilities and 39.4% in patients without incompatibilities (p = 0.018). The respective relapse rates of patients with CD62L, HA-1, CD31 exon 563, CD31 exon 125 and 49b incompatibilities were 6.1%, 14.3%, 11.7%, 20% and 40%. Only patients with CD62L incompatibilities showed a lower relapse rate than patients who received a compatible graft. Since there was no difference between patients with and without incompatibilities as far as the appearance of acute GVHD (> or = 2) was concerned, we conclude that the polymorphic CD62L molecule contributes to graft versus-leukemia rather than the development of GVHD after HLA-identical stem cell transplantation. PMID- 15359911 TI - [Autologous peripheral blood stem cell transplantation for Japanese multiple myeloma patients: results of a feasibility study]. AB - A feasibility study on high-dose therapy with autologous peripheral blood stem cell transplantation (HDT/PBSCT) was performed in Japanese patients with multiple myeloma (MM). Twenty evaluable patients younger than 65 years old with stage II/III MM were enrolled in this study. Three courses of VAD were used as initial chemotherapy. High-dose etoposide or cyclophosphamide followed by G-CSF was used for PBSCH, and 1.2-89.3 (median 23.4) x 106/kg of CD34+ cells were collected. Single (11 patients) or tandem (9 patients) HDT with melphalan (MEL) 200 mg/m2 or MEL 140 mg/m2 plus TBI 10 Gy were performed. The incidence of grade 4 toxicity (COG) was 10% and treatment-related mortality was 5%. Complete response and tumor reduction of more than 75% were obtained in 4 (21%) and 16 (84%) out of 19 patients, respectively. The actuarial 3-year overall survival (OS) and event-free survival (EFS) after PBSCT/HDT were 65.6% and 22.0%, respectively. The median EFS duration was 18 months. These preliminary results indicated that HDT/PBSCT is feasible for Japanese MM patients. A prospective randomized clinical trial will be required to assess the efficacy. PMID- 15359912 TI - [Clinical significance of detecting strong bcl-2 protein expression with flow cytometry in follicular lymphoma]. AB - The expression of cell surface antigens and bcl-2 protein was examined with flow cytometry in 30 lymphoma samples including 28 lymph nodes and others, from January, 2001 to May, 2003. Cases studied included the following: follicular lymphoma (FL), 11; diffuse large B-cell lymphoma (DLBCL), 12; diffuse small B cell lymphoma, 1; mantle cell lymphoma, 1; MALT lymphoma, 1; precursor-B lymphoblastic lymphoma, 1; T-lymphoblastic lymphoma, 1; blastic NK-cell lymphoma, 1; and unknown, 1. In addition, cytogenetic analysis, fluorescence in situ hybridization (FISH), and a real-time polymerase chain reaction assay were performed. Strong bcl-2 expression was observed in 91% of FL, and 17% of DLBCL. On the other hand, strong bcl-2 expression was found in all 9 cases of FL with t(14;18), all two cases of DLBCL with t(14;18), one unknown case with t(14;18), and one FL case with t(11;14). Our results support that strong bcl-2 expression as demonstrated with flow cytometry is correlated with FL (p<0.0001), and lymphomas with the t(14;18) (p<0.0001). PMID- 15359914 TI - [Pleural involvement in the course of chronic myelomonocytic leukemia and the development of multiple colonic perforation due to leukemic infiltration in the acute leukemia phase]. AB - A 68-year-old man with chronic myelomonocytic leukemia (CMML) was initially treated with hydroxyurea with subsequent stable disease. In the time course, he developed bilateral pleuritis accompanied by leukocytosis and spiking fever. Cytologic analysis of the pleural effusion revealed abundant mature granulocytes and monocytes. He was treated with intravenous or oral etoposide with consequent resolution of the pleuritis, indicating the pleural involvement of CMML. Three months later, he developed hepatomegaly and became febrile. One month thereafter, the CMML transformed to acute myeloid leukemia, and the patient developed massive bloody stools associated with epigastric pain and leukocytosis. A gastrofiberscopic examination showed multiple bleeding gastric ulcers. The bleeding ulcers were treated with the clipping procedure; however, the bloody stools continued. Although intravenous etoposide was effective for the leukocytosis and hepatomegaly, the treatment did not improve the bloody stools. The patient finally died of panperitonitis. The autopsy showed multiple ulcers of the transverse colon, some of which were perforated. Microscopically, the ulcerated areas were densely infiltrated with leukemic cells predominantly consisting of immature monocytes and granulocytes. This patient may be the first reported case of CMML complicated by colonic perforation due to leukemic infiltration. PMID- 15359913 TI - [Current status of treatment for patients with idiopathic thrombocytopenic purpura in the Hokkaido area (evaluation of Helicobacter pylori eradication)]. AB - Treatment guidelines for patients with idiopathic thrombocytopenic purpura (ITP) have been changed recently due to the clinical application of Helicobacter pylori (H. pylori) eradication but there has been no detailed multi-center analysis of the hematological effects of H. pylori eradication. The Clinical Hematology Forum consists of 11 large hematological departments and divisions in the Hokkaido area. We sent questionnaires to these 11 hematological departments and divisions in March 2003 to obtain information on current treatment strategies for patients with ITP and hematological results after the eradication of H. pylori. Questionnaires were returned by 9 (81.8%) of the 11 departments. Doctors in all hospitals had experience in diagnosis and treatment of H. pylori infection. Diagnostic examinations for H. pylori infection were performed in 54.3% of the registered cases. H. pylori infection was detected in 68.1% of the examined cases, and eradication treatment was performed in 87.7% of H. pylori-positive patients. H. pylori was eradicated in 52 (83.9%) of the 62 patients in whom the results of treatment could be evaluated. Among the patients whose platelet counts were less than 10.0 x 10(4)/microl, platelet recovery was observed in 48.8% of cases with successful eradication, a percentage similar to previously reported percentages in Japan. There was no prognostic factor to predict good responders before eradication treatment. Since the side effects of eradication treatment, including gastrointestinal symptoms and skin eruptions, were not serious, this method might become a front-line treatment for patients with ITP. Patient selection for eradication as an up-front treatment, analysis of the pathophysiology of platelet recovery after eradication and long-term effects should be investigated to make new treatment guidelines for newly diagnosed patients with ITP. PMID- 15359915 TI - [Mantle cell lymphoma markedly infiltrated into adrenal glands with adrenal insufficiency]. AB - A 66-year-old male was admitted to our hospital complaining of bilateral hypochondrial pain, back pain and loss of weight in May, 2002. Superficial lymph nodes were not palpable on admission. The leukocyte count was 3430/microl, hemoglobin concentration, 13.0g/dl, and platelet count, 174000/microl. LDH, soluble IL-2 receptor, ACTH and cortisol values were out of the normal range (LDH 1368IU/l, sIL-2R 2630U/ml, ACTH 132pg/ml, cortisol 7.4microg/dl). Abdominal CT scan showed bilateral adrenal masses, and abnormal uptake of Ga-scintigraphy was seen correspondent with the bilateral adrenal masses. The histological diagnosis of bilateral adrenal masses cannot be performed because of the bleeding tendency, but atypical cells were observed in the patient's bone marrow aspirate. Surface marker analysis of atypical cells showed CD5+, cyclin D1+, CD19+, CD20+ and HLA DR+. From these results we diagnosed this case as a mantle cell lymphoma (stage IV B) markedly infiltrated into the adrenal glands with adrenal insufficiency. The bilateral adrenal masses dramatically reduced in size after CHOP chemotherapy with hydrocortisone supplementation. We report on the present case and summarize the reports of adrenal grand-infiltrating lymphomas. PMID- 15359916 TI - [Granulocyte transfusions for severe infections prior to allogeneic hematopoietic stem cell transplantation]. AB - The efficacy and safety of granulocyte transfusions were evaluated in two acute lymphoblastic leukemia patients for the control of severe infections (cervical cellulitis, sepsis) prior to hematopoietic stem cell transplantation. One patient received 6 transfusions and the other 2 transfusions. The donors were given subcutaneous granulocyte-colony stimulating factor plus oral dexamethasone/betamethasone 12 hours before the scheduled collection. Granulocytes were obtained by standard leukapheresis procedures utilizing hydroxyethyl starch with processing of 7 liters of blood. The yield was 3.2-10.7 x 10(10) (0.7-2.1 x 10(9)/kg of recipient) granulocytes. Post-transfusion increases of peripheral blood neutrophil counts in the following morning were 300 to approximately 6,900/ml. Infections resolved and successful engraftment was obtained in both patients after the transplants. No severe adverse reactions were observed. These findings suggest that granulocyte transfusions are useful for control of severe infections prior to allogeneic hematopoietic stem cell transplantation. PMID- 15359918 TI - [Cladribine monotherapy for patients with relapsed or refractory indolent non Hodgkin lymphoma]. AB - Cladribine is a purine analogue that is resistant to degradation by adenosine deaminase. We describe the efficacy of cladribine monotherapy in 8 patients with relapsed or refractory indolent non-Hodgkin lymphoma. The median age of the patients was 57 years. All patients were given 1-3 courses of cladribine monotherapy at 0.09 mg/kg/day continuous infusion for 7 days. Although all patients had been treated with rituximab and CHOP-like regimens, the response rate was excellent (85.7%). Thus we consider that cladribine is the first treatment of choice in the patients with relapsed or refractory indolent lymphoma. PMID- 15359917 TI - [A family of acute intermittent porphyria]. AB - A 62-year-old man who had twice received laparotomies for abdominal pain of unknown origin was admitted to our hospital with acute abdominal pain. His family history of acute intermittent porphyria (AIP) suggested that it arose from acute porphyria. We treated the patient with 5% glucose solution by i.v. drip infusion and his abdominal pain improved rapidly. Diagnosis of AIP was established by the demonstration of reduced erythrocyte porphobilinogen deaminase (PBGD) activity and a point mutation (CAG --> CGG) in a splicing site in intron 10/exon 11 in the PBGD gene by DNA analysis. For screening of AIP carriers in his family, we measured erythrocyte PBGD activity. Four of his seven children were successfully diagnosed as AIP carriers. This is the ninth AIP family report, in which a mutation in the PBGD gene was revealed by DNA analysis. PMID- 15359919 TI - Gene expression imaging with radiolabeled peptides. AB - An approach to image radiolabeled peptide localization at tumor sites by inducing tumor cells to synthesize membrane expressed human somatostatin receptor subtype 2 (hSSTr2) with a high affinity for radiolabeled somatostatin analogues is described. The use of gene transfer technology to induce expression of high affinity membrane hSSTr2 can enhance the specificity and degree of radiolabeled peptide localization in tumors. Employing this strategy, induction of high levels of hSSTr2 expression with selective tumor uptake of radiolabeled peptides was achieved in both subcutaneous non-small cell lung cancer and intraperitoneal ovarian cancer mouse human tumor xenograft models. The features of this genetic transduction imaging approach are: (1) constitutive expression of a tumor associated receptor is not required; (2) tumor cells are altered to express a new target receptor or increased quantities of a constitutive receptor at levels which may significantly increase tumor targeting of radiolabeled peptides compared to uptake in normal tissues; (3) gene transfer can be accomplished by local or regional injection of adenoviral vectors; (4) it is feasible to target adenovirus vectors to tumor cells by modifying adenoviral tropism (binding) or by the use of tumor-specific promoters such that the hSSTr2 will be specifically expressed in the desired tumor; and (5) this technique can be used to image expression of a second therapeutic gene. PMID- 15359920 TI - Water-pipe smoking effects on pulmonary permeability using technetium-99m DTPA inhalation scintigraphy. AB - OBJECTIVE: Although extensive work has been done on cigarette smoking and its effects on pulmonary function, there are limited number of studies on water-pipe smoking. The effects of water-pipe smoking on health are not widely investigated. The aim of this study was to determine the effects of water-pipe smoking on pulmonary permeability. METHODS: Technetium-99m DTPA inhalation scintigraphy was performed on 14 water-pipe smoker volunteers (all men, mean age 53.7 +/- 9.8) and 11 passive smoker volunteers (1 woman, 10 men, mean age 43.8 +/- 12). Clearance half-time (T 1/2) was calculated by placing a monoexponential fit on the time activity curves. Penetration index (PI) of the radioaerosol was also calculated. RESULTS: PI was 0.58 +/- 0.14 and 0.50 +/- 0.12 for water-pipe smokers (WPS) and passive smokers (PS) respectively. T 1/2 of peripheral lung was 57.3 +/- 12.7 and 64.6 +/- 13.2 min, central airways was 55.8 +/- 23.5 and 80.1 +/- 35.2 min for WPS and PS, respectively (p < or = 0.05). FEV1/FVC% was 82.1 +/- 8.5 (%) and 87.7 +/- 6.5 (%) for WPS and PS, respectively (0.025 < p < or = 0.05). CONCLUSIONS: We suggest that water-pipe smoking effects pulmonary epithelial permeability more than passive smoking. Increased central mucociliary clearance in water-pipe smoking may be due to preserved humidity of the airway tracts. PMID- 15359921 TI - Methionine positron emission tomography for differentiation of recurrent brain tumor and radiation necrosis after stereotactic radiosurgery--in malignant glioma. AB - OBJECT: Following stereotactic radiosurgery (SRS), we examined how to differentiate radiation necrosis from recurrent malignant glioma using positron emission tomography (PET) with 11C-methionine (Met). METHODS: Met-PET scans were obtained from 11 adult cases of recurrent malignant glioma or radiation injury, suspected on the basis of magnetic resonance images (MRI). Patients had previously been treated with SRS after primary treatment. PET images were obtained as a static scan of 10 minutes performed 20 minutes after injection of Met. We defined two visual grades (e.g., positive or negative Met accumulation). On Met-PET scans, the portion of the tumor with the highest accumulation was selected as the region of interest (ROI), tumor-versus-normal ratio (TN) was defined as the ratio of average radioisotope counts per pixel in the tumor (T), divided by average counts per pixel in normal gray matter (N). The standardized uptake value (SUV) was calculated over the same tumor ROI. Met-PET scan accuracy was evaluated by correlating findings with subsequent histological analysis (8 cases) or, in cases without surgery or biopsy, by the subsequent clinical course and MR findings (3 cases). RESULTS: Histological examinations in 8 cases showed viable glioma cells with necrosis in 6 cases, and necrosis without viable tumor cells in 2 cases. Three other cases were considered to have radiation necrosis because they exhibited stable neurological symptoms with no sign of massive enlargement of the lesion on follow-up MR after 5 months. Mean TN was 1.31 in the radiation necrosis group (5 cases) and 1.87 in the tumor recurrence group (6 cases). Mean SUV was 1.81 in the necrosis group and 2.44 in the recurrence group. There were no statistically significant differences between the recurrence and necrosis groups in TN or SUV. Furthermore, we made a 2 x 2 factorial cross table (accumulation or no accumulation, recurrence or necrosis). From this result, the Met-PET sensitivity, specificity, and accuracy in detecting tumor recurrence were determined to be 100%, 60%, and 82% respectively. In a false positive-case, glial fibrillary acidic protein (GFAP) immunostaining showed a positive finding. CONCLUSION: There were no significant differences between recurrent malignant glioma and radiation necrosis following SRS in Met-PET. However, this study shows Met-PET has a sensitivity and accuracy for differentiating between recurrent glioma and necrosis, and presents important information for developing treatment strategies against post radiation reactions. PMID- 15359922 TI - Quantitative PET cerebral glucose metabolism estimates using a single non arterialized venous-blood sample. AB - The purpose of this study is to develop a method of quantitating the cerebral metabolic rate of glucose (CMRglc) by positron emission tomography using a population-based heated venous curve and one-point sampling from a non-heated vein, i.e. that can avoid arterial puncture. METHODS: We conducted this study on 17 subjects with a mean age of 61 +/- 9 years. A time-concentration curve as an input function was obtained by sampling 24 blood samples, from the heated left hand vein, one before and the others after intravenous injection of 259 MBq of F 18-fluorodeoxyglucose into the right cubital vein. A non-heated venous sample was also obtained from the right cubital vein. RESULTS: The population-based input function was calculated by averaging time-concentration curves from the first 7 subjects. A single sample obtained from 10 other subjects from 7.5 to 20 minutes and 35 and 40 minutes after injection predicted input function well with an error of less than 4.5%. The radioactivity in the non-heated 40 minutes' sample was 1.7 +/- 2.9% higher than in the heated vein. When we calibrated the population-based curve using the non-heated venous samples at 40 minutes in 10 subjects, the calculated CMRglc values were 1.3 +/- 5.4% lower than the actual values. CONCLUSIONS: Non-heated venous one-point sampling and the population-based curve can decrease the complexity of the procedures and the manpower required, and also make the FDG study less invasive, without a significant increase in measurement error. PMID- 15359923 TI - Alveolar epithelial permeability in patients with primary spontaneous pneumothorax as determined by Tc-99m DTPA aerosol scintigraphy. AB - PURPOSE: Primary spontaneous pneumothorax (PSP) occurs subsequent to a disruption in the continuity of visceral pleura and escape of air into the pleural space. The cause of PSP is most often the rupture of subpleural blebs or bullae. It is usually difficult to detect evidence of pulmonary pathology. The purposes of the present study were (1) to investigate the changes of pulmonary alveolar epithelial permeability in patients with PSP as determined by Tc-99m DTPA aerosol lung scintigraphy, (2) to assess whether or not some differences exist between apical and basal parts of the lungs, and (3) to determine the relationship between the clearance rate of Tc-99m DTPA and the PFT results, the recurrence rate of PSP, and the percentage of pneumothorax in affected lung. MATERIAL AND METHODS: Thirteen PSP patients (two females, 11 males; mean age 32.5 +/- 11.8 years) with normal chest X-ray were studied. Thirteen healthy non-smoking volunteers (1 female, 12 males; mean age, 35.8 +/- 10 years) were selected as a control group. Tc-99m DTPA aerosol lung scintigraphy and PFT were performed in all patients and controls. Clearance rates (%/min) of Tc-99m DTPA aerosol in right and left lung field, and apical and basal parts of each lung were calculated from dynamic images for 15 min. RESULTS: There was no significant difference (p > 0.05) between patients and controls, or between apical and basal parts of each lung. No correlation was found between the clearance rate of Tc-99m DTPA and PFT results, the recurrence rate of PSP, or the percentage of pneumothorax. CONCLUSION: This study demonstrates that pulmonary epithelial permeability is not altered in PSP patients; the clearance rate of Tc-99m DTPA shows no difference between apical and basal parts of each lung. PMID- 15359924 TI - Value of axillary lymphoscintigraphy in patients with operated breast carcinoma. AB - PURPOSE: To evaluate axillary dissection with axillary lymphoscintigraphy (ALS) in postoperative patients with breast carcinoma and its role in adjuvant radiotherapy (RT). Additionally, to define axillary dissection as complete and incomplete with ALS and to correlate it with the number of removed lymph nodes. MATERIAL AND METHODS: In the last two years, 121 women were studied four weeks after operation. Bilateral second interdigital subcutaneous injections were performed for ALS. Complete and incomplete axillary dissection were interpreted according to the number of surgically removed lymph nodes. ALS was interpreted as complete if no accumulation was shown. RESULTS: There was a good correlation between the number of surgically removed lymph nodes and complete and incomplete interpretation on ALS (p < 0.004). The number of removed lymph nodes was equal to or greater than 15 in 72% patients with complete dissection according to ALS. Of 48 patients with surgically incomplete axillary dissection, 18 (38%) showed no accumulation in the axillary region, while 25 of 68 (37%) patients with surgically complete dissection showed accumulation in the axillary region and were interpreted as incomplete according to ALS. Indication of RT was changed after ALS in patients with 1 to 3 involved lymph nodes. While RT was not considered in 12 of these patients before ALS, they were included in RT planning. On the other hand, 17 patients, considered for RT previously, were excluded from RT planning after ALS. CONCLUSION: Evaluation of axillary dissection with ALS especially in suspicious patients with 1 to 3 lymph node metastases might prevent unnecessary morbidity and can be useful in selecting patients who truly need axillary irradiation. PMID- 15359925 TI - Attenuation correction using asymmetric fanbeam transmission CT on two-head SPECT system. AB - For transmission computed tomography (TCT) systems using a centered transmission source with a fan-beam collimator, the transmission projection data are truncated. To achieve sufficiently large imaging field of view (FOV), we have designed the combination of an asymmetric fan-beam (AsF) collimator and a small uncollimated sheet-source for TCT, and implemented AsF sampling on a two-head SPECT system. The purpose of this study is to evaluate the feasibility of our TCT method for quantitative emission computed tomography (ECT) in clinical application. Sequential Tc-99m transmission and Tl-201 emission data acquisition were performed in a cardiac phantom (30 cm in width) with a myocardial chamber and a patient study. Tc-99m of 185 MBq was used as the transmission source. Both the ECT and TCT images were reconstructed with the filtered back-projection method after scatter correction with the triple energy window (TEW) method. The attenuation corrected transaxial images were iteratively reconstructed with the Chang algorithm utilizing the attenuation coefficient map computed from the TCT data. In this AsF sampling geometry, an imaging FOV of 50 cm was yielded. The attenuated regions appeared normal on the scatter and attenuation corrected (SAC) images in the phantom and patient study. The good quantitative accuracy on the SAC images was also confirmed by the measurement of the Tl-201 radioactivity in the myocardial chamber in the phantom study. The AsF collimation geometry that we have proposed in this study makes it easy to realize TCT data acquisition on the two-head SPECT system and to perform quantification on Tl-201 myocardial SPECT. PMID- 15359926 TI - Assessment of cerebral hemodynamics in childhood moyamoya disease using a quantitative and a semiquantitative IMP-SPECT study. AB - BACKGROUND: We evaluated the cerebral hemodynamics in childhood moyamoya disease patients before and after surgery to assess both surgical indication and the effect of revascularization using single photon emission computed tomography (SPECT) study with N-isopropyl-p-123I-iodoamphetamine (IMP). We compared results of quantitative and semi-quantitative SPECT studies to determine parameters by the semi-quantitative method to define severe hemodynamic ischemia. METHODS: There were 14 pediatric patients with moyamoya disease who suffered transient ischemic attacks (TIAs) in the anterior circulation. Before and after surgical revascularization by STA-MCA bypass and encephalomyosynangiosis (EMS), quantitative IMP-SPECT studies using the autoradiographic method (IMP-ARG method) were performed. Resting regional cerebral blood flow (rCBF) and regional vascular reserve (rVR) were measured in bilateral cortical territories (ROI) and cerebellum. Semi-quantitative parameters were calculated from the ratio of ROI counts to the dominant cerebellar counts (ROI/Ce ratio) at resting and acetazolamide-activated conditions. RESULTS: Before surgery, the mean resting rCBF and rVR in bilateral ACA and MCA territories were less than 40 ml/100 g/min and less than 10%, respectively, indicating severe hemodynamic ischemia. Except for the ACA territories, both the mean resting rCBF and mean rVR values in the entire cortex increased significantly after surgery (p < 0.05). By semi quantitative studies, before surgery, the mean resting and acetazolamide activated ROI/Ce ratios in bilateral ACA and MCA territories were less than 0.90 and 0.80, respectively. The mean resting and acetazolamide-activated ROI/Ce ratios increased significantly in the MCA territory after surgery. Severe hemodynamic ischemia, which categorized by the quantitative thresholds (resting rCBF < 40 ml/100 g/min and rVR < 10%) was diagnosed by the semi-quantitative thresholds (resting ROI/ Ce ratio < 0.90 and acetazolamide-activated ROI/Ce ratio < 0.85), the sensitivity and specificity of which were 87.5% and 90.9%, respectively. CONCLUSIONS: The cerebral hemodynamics in childhood moyamoya disease was improved entirely after surgery. Severe hemodynamic cerebral ischemia was diagnosed by not only quantitative but also semi-quantitative IMP-SPECT studies. PMID- 15359927 TI - Quantitative assessment of truncal FDG-PET examination with postinjection transmission scan--comparison with preinjection transmission scan. AB - The purpose of this study was to assess the quantitative accuracy of truncal FDG PET with a postinjection transmission scan. METHODS: Ten subjects with lung cancer were recruited for this study. Prior to the emission scan, a transmission scan was performed for 10 min. All subjects received 370 MBq of intravenous administration of FDG prior to a 60-min emission scan. Immediately following the emission scan, a postinjection transmission scan was performed. Emission data from 40 to 60 min postinjection were reconstructed with either pre- or postinjection transmission data and converted to a standardized uptake value (SUV) image. On each SUV image, 5 regions of interest were placed and regions of interest on the SUV image with a postinjection transmission scan (SUVpost) were plotted against those with preinjection transmission (SUVpre), and a regression line was generated. Using the slope and Y-intercept of the regression line, percent error of estimation of the SUV was calculated based on the following equation: % error = ISUVpre - SUVpostI x 100/SUVpre. RESULTS: In the low SUV area (SUV = 1), the averaged percent error was 9.4 +/- 12.0% (mean +/- SD), whereas in the high SUV area (SUV = 10), the averaged percent error was 2.8 +/- 3.1%. The least percent error was 1.8 +/- 1.8% (SUV = 3.8) in this study. CONCLUSION: In the study on truncal FDG PET with postinjection transmisson scan, the quantitative accuracy was preserved and the method is clinically available. PMID- 15359928 TI - 5-[123I]Iodo-A-85380: assessment of pharmacological safety, radiation dosimetry and SPECT imaging of brain nicotinic receptors in healthy human subjects. AB - Recently, 5-[123I]iodo-3-(2(S)-azetidinylmethoxy)pyridine ([123I]5IA) was developed as a ligand for imaging the nicotinic acetylcholine receptor (nAChR) in human brain using single photon emission computed tomography (SPECT). In the present study, the toxicity and radiation absorbed dose of [123I]5IA were investigated. Behavior and physiological parameters were examined in mice and rats after administration of 5IA. There were no changes in these parameters in animals administered 1 microg/kg of 5IA or less, indicating that the no observed effect level (NOEL) of 5IA was 1 microg/kg. [123I]5IA was then administered to healthy human subjects and serial whole-body images were acquired over 24 hr. Initially, high levels of radioactivity were observed in the liver and urinary bladder and moderate levels in the lungs, kidneys, and brain. Whole brain activity at 1 hr was 4.6 +/- 0.4% of the injected dose and this value gradually decreased with time. The majority (-75%) of the radioactivity was excreted in urine within 24 hr, and less than 1% remained in all organs tested. The biological half-life of [1231]51A averaged 7.2 +/- 4.0 hr. Based on the biodistribution data, radiation absorbed doses were estimated using MIRDOSE 3.1 software with the dynamic bladder model and the ICRP gastrointestinal (GI) tract model. Consequently, the effective dose equivalent was estimated to be 30 +/- 1.4 microSv/MBq, which is an acceptable radiation burden. Having determined the safety of this compound, we performed SPECT imaging in a healthy human subject using 171 MBq of [123I]5IA. SPECT images clearly revealed a cerebral distribution of radioactivity that was consistent with the known distribution of central nAChRs in humans. These results suggest that [123I]5IA is a promising ligand for imaging nAChRs in humans, with an acceptable dosimetry and pharmacological safety at the dose required for adequate SPECT imaging. PMID- 15359929 TI - Multifactorial analysis on the short-term side effects occurring within 96 hours after radioiodine-131 therapy for differentiated thyroid carcinoma. AB - OBJECTIVE: This study was performed to clarify factors that might influence short term side effects occurring within 96 hours after administration of 131I for patients with thyroid carcinoma. METHODS: In 71 patients with differentiated thyroid carcinoma, short-term side effects including gastrointestinal complaints, salivary gland swelling with pain, change in taste and headache were retrospectively analyzed. All patients were given domperidone for prevention of gastrointestinal complaints and advised to consume sour foods to promote discharge of radioiodine from the salivary glands. Selected factors possibly affecting the incidence of side effects were dose per body weight, TSH, effective half-life of 131I, sex, age, 131I accumulation into the stomach and salivary glands, and edema prior to radioiodine administration. The factors were evaluated by multivariate analyses. RESULTS: Incidence of gastrointestinal complaints, salivary gland swelling with pain, change in taste and headache was 65.2%, 50.0%, 9.8% and 4.4%, respectively. In gastrointestinal complaints, the incidence of appetite loss, nausea and vomiting was 60.9%, 40.2% and 7.6%, respectively. The gastrointestinal complaints increased significantly in the patients dosed above 55.5 MBq/kg and with TSH elevation. For salivary gland swelling with pain, female patients displayed a significantly higher incidence than males. No statistically significant factors were detected for change in taste or headache. CONCLUSIONS: Significant factors influencing short-term side effects were dose per body weight and TSH values for gastrointestinal complaints, and female sex for salivary gland swelling with pain. Our preliminary experience suggests that the most frequent gastrointestinal complaints can be prevented with ramosetron. PMID- 15359930 TI - FDG-PET imaging in duodenal cancer. AB - F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) imaging is a useful modality in the detection of various tumors, including colon cancer and gastric cancer. We evaluated a patient with duodenal cancer with multiple metastases including brain metastases using FDG-PET imaging. It revealed multiple tumor uptake in the brain, clavicular fossa, mediastinum, right adrenal gland and duodenum. These results suggest that FDG-PET imaging may be useful in detecting the primary and metastatic lesions of duodenal adenocarcinoma. PMID- 15359931 TI - Usefulness of asialoglycoprotein receptor imaging for the evaluation of liver metastasis of neuroblastoma. AB - Neuroblastoma, derived from the neural crest ectoderm, is the most common type of solid abdominal mass seen in infancy. The diagnosis, staging, and follow-up of neuroblastoma are often performed using metaiodobenzylguanidine (MIBG) imaging. However, the evaluation of liver metastasis by this method is complicated by the normal physiological uptake of MIBG by the liver. The asialoglycoprotein receptor is a hepatic cell-surface receptor specific for galactose-terminated glycoprotein, and 99mTc-DTPA-galactosyl human serum albumin (GSA) accumulates selectively in hepatic cells. Here, we report a case of congenital neuroblastoma with liver metastasis in which GSA scans were useful for differentiation between normal and metastatic sites in the liver. PMID- 15359932 TI - [Mechanisms involved in the activation of the protein C pathway in sepsis]. PMID- 15359933 TI - [PROWESS: description and key results]. PMID- 15359934 TI - [Hemorrhagic risk of drotrecogin alfa (activated) in severe sepsis]. PMID- 15359935 TI - [The FDA's questions]. PMID- 15359936 TI - [Question of the European Agency of Drugs (EMEA): interaction between drotrecogin alfa (activated) and heparin in prophylactic dose]. PMID- 15359937 TI - [Medical-economic evaluation of severe sepsis]. PMID- 15359938 TI - [Severe sepsis: perspectives]. PMID- 15359939 TI - [Coagulopathy in severe sepsis: is there anything new?]. PMID- 15359940 TI - [Which monitoring and follow-up in organ dysfunctions during severe sepsis?]. PMID- 15359941 TI - [Surgical patients with severe sepsis: first results of the PROWESS study. Demographic data]. PMID- 15359942 TI - [Sepsis in the surgical setting: indication for activated C protein]. PMID- 15359943 TI - [Xigris and corticoids: what are the perspectives for the patient with severe sepsis?]. PMID- 15359944 TI - [Clinical experience with xigris]. PMID- 15359945 TI - [Can genetics guide or modify the management of severe sepsis?]. PMID- 15359946 TI - [Clinical approach to the identification of patients suffering from severe sepsis with several organ failures for the administration of drotrecogin alfa (activated): from the PROWESS study to clinical practice]. PMID- 15359947 TI - [Hyperkalemia. Main symptoms: muscle weakness, cardiac arrhythmias]. PMID- 15359948 TI - [How gender and communication style of physicians affect patient satisfaction: the little difference]. AB - Patients are particularly satisfied with the medical consultation when physicians adopt a patient-oriented communication style which is characterized by emotional participation, exploring the meaning of a disease for the life of the patient, and including the patient in decision making. Female physicians are much more likely to adopt such a patient-oriented communication style than male physicians. As a consequence, patients should be more satisfied after consultations with female than with male physicians. However, research shows that there is no difference: patients are equally satisfied with female and male physicians. This article sheds light on this paradox and explains it based on gender role stereotypes and patient expectations. PMID- 15359949 TI - [Ovarian torsion in childhood]. AB - A six year old afebrile girl was referred to the emergency department with abdominal pain in the right lower region. A retrouterine partially cystic lesion with an inhomogene echogenity was demonstrated by transabdominal ultrasound. Pelvic MR showed an enlarged right-sided ovary and a thickened vascular pedicle as well as few adjacent free fluid. Diagnosis of ovarian torsion was confirmed intraoperatively. Conservative ovary-sparing treatment was not possible, so right sided adnexectomy followed. Histopathologically a haemorrhagic infarction of the ovary and the tube was seen. No signs of malignancy and no tumor could be detected. PMID- 15359950 TI - [Subacute thoracic pain]. AB - A 75-year old woman with essential hypertension presented two days after the onset of a sudden and strong thoracic pain. The pain did neither increase during breathing nor decrease after the ingestion of nitroglycerine, and could not be triggered by physical manipulation of the thoracic wall. Electrocardiogram showed a left bundle branch block, chest X-ray showed a widened upper mediastinum. Rupture/dissection of an aberrant right subclavian artery (a. lusoria) could be diagnosed by computed tomography. Successful implantation of an endovascular stent-graft was carried out. Evaluation of (sub)acute thoracic pain should include two-plane chest X-ray and, in case of a widened mediastinum, further investigation by computed tomography. In case of highly suspected rupture/dissection of an intrathoracic artery, a specific imaging procedure such as computed tomography is crucial. PMID- 15359951 TI - [69-year-old patient with anemia, hypothyroidism and transferrin saturation]. PMID- 15359952 TI - [The contradictory qualities of Podophyllum]. PMID- 15359953 TI - Mystery piglet deaths prompt animal health investigation. PMID- 15359955 TI - Exotic animal disease bulletin. PMID- 15359954 TI - The veterinary profession--a global perspective. PMID- 15359956 TI - Welcoming new employees--maximising your chances of a good return on your investment. PMID- 15359957 TI - Bain fallon. Equine reproduction and horsing around. PMID- 15359958 TI - Chip shot--right of reply. PMID- 15359959 TI - AVA international veterinary aid. PMID- 15359960 TI - Australian veterinarians who work with horses: views of the future. AB - OBJECTIVE: To give an account of the views held by Australian veterinarians who work with horses on the future of their professional field. METHOD: Questionnaires were mailed to 866 veterinarians who had been identified as working with horses, and 87% were completed and returned. Data were entered onto an Excel spreadsheet, and analysed using the SAS System for Windows. RESULTS: Their future prospects were believed to be very good or excellent by >60% of equine veterinarians but by only 30% of mixed practitioners seeing < 10% horses. The main factors believed likely to affect these prospects were the strength of the equine industries and the economic climate affecting horse owners, followed by the encroachment of cities into areas used for horses, competition from other veterinarians including specialist centres and from non-veterinary operators, and their ability to recruit and retain veterinarians with interest, experience and skill with horses. Urban encroachment, competition and recruitment were especially important for those seeing few horses. Concerns were also expressed about the competence and ethical behaviour of other veterinarians, the physical demands and dangers of horse work, the costs of providing equine veterinary services and of being paid for them, the regulatory restrictions imposed by governments and statutory bodies, the potential effects of litigation, and insurance issues. For many veterinarians in mixed practice these factors have reduced and are likely to reduce further the number of horses seen, to the extent that they have scant optimism about the future of horse work in their practices. CONCLUSION: Economic and local factors will result in an increasing proportion of equine veterinary work being done in specialised equine centres, and the future of horse work in many mixed practices is, at best, precarious. A key factor influencing future prospects will be the availability of competent veterinarians committed to working with horses. PMID- 15359961 TI - Retrospective study of old world screw-worm fly (Chrysomya bezziana) myiasis in 59 dogs in Hong Kong over a one year period. PMID- 15359962 TI - Nonfatal splenic haematoma and pancytopenia in an ass. AB - This report describes a case of nonfatal idiopathic splenic haematoma and pancytopenia in a female ass, a condition that has not been previously reported in this species. A 12-year-old female ass weighing 225 kg was presented with signs of anorexia, depression and prominent abdominal distension. At the onset of clinical signs there was evidence of anaemia, leucopenia and thrombocytopenia. A large mass was palpated per rectum. The mass was confluent with the spleen and extended across the entire width of the abdomen. Percutaneous transabdominal ultrasonography and laparoscopy under general anaesthesia confirmed the diagnosis of splenic haematoma. The ass was treated initially with intravenous polyionic fluids; vitamins, trace elements and aminoacids; clanobutin; and penicillin and gentamicin, and was transfused with whole blood. After 5 days of hospitalisation the ass appeared brighter, and after 10 days the animal was discharged to recuperate at pasture. Six months later the animal's demeanour and appetite had improved. Repeat ultrasonographic examination showed persistence of the mass, but its thickness had decreased by 2 to 3 cm. A small mineralised area suggesting organisation of the haematoma was also detected. The haematological parameters had returned to normal values and the ass was clinically normal. PMID- 15359963 TI - Haemoperitoneum secondary to granulosa cell tumour in two mares. PMID- 15359964 TI - Antibiotic responsive ulcerative dermatoses in German Shepherd Dogs with mucocutaneous pyoderma. AB - Mucocutaneous pyoderma is a disease of unknown aetiology affecting mucocutaneous skin and is responsive to antibacterial therapy. It is reported to affect the lips, nasal planum, nares, perioral skin and less commonly, the eyelids, vulva, prepuce and anus. Three cases of mucocutaneous pyoderma are presented. Two of the cases showed ulcerative lesions in the inguinal and axillary regions in addition to more typically reported lesions. Two of the dogs had concurrent atopic dermatitis and the third had clinical signs suggestive of hyper-sensitivity disease. The clinical and histopathological features, differentiation of mucocutaneous pyoderma from discoid lupus erythematosus, and long-term management of mucocutaneous pyoderma are discussed. PMID- 15359965 TI - Review of possible links between the clinical expression of paratuberculosis and deficiency of macro and micronutrients. AB - Published information on aspects of the epidemiology of paratuberculosis was reviewed to examine the hypothesis that clinical expression of disease may be induced by deficiencies or imbalances of micronutrients. The progression of paratuberculosis and the development of clinical signs, although innately linked with the process of becoming infected with Mycobacterium avium subsp paratuberculosis, are mostly controlled by factors other than those predisposing to the acquisition of the organisms. There is strong evidence in the literature reviewed to suggest that soil acidification, excesses of iron and molybdenum, and deficiencies in copper and selenium and possibly other minerals and trace elements have a role in the disease process. The fact that no single nutritional supplement has been consistently found to produce clinical recovery suggests that the causal role of these factors is complex and may vary depending upon the geochemical environment. Altering the nutritional environment by appropriate trace element supplementation and liming, as well as decreasing the reliance on shallow-rooted pastures needing frequent fertiliser applications may assist in controlling the clinical expression of paratuberculosis. Such measures could produce a significant decrease in the incidence of clinical expression across flocks and herds. PMID- 15359966 TI - Nested PCR on blood and milk for the detection of Mycobacterium avium subsp paratuberculosis DNA in clinical and subclinical bovine paratuberculosis. AB - OBJECTIVE: To determine the potential of PCR on blood and milk to detect cattle infected with Mycobacterium avium subsp paratuberculosis. PROCEDURE: A nested PCR method probing for IS900 was developed and compared to ELISA serology in 11 clinically infected and 46 subclinically infected, lactating Holstein cows from a herd with confirmed paratuberculosis (Johne's disease). RESULTS: When compared to serum ELISA the nested blood- and milk PCRs were equal in identifying DNA from clinically infected animals. The PCR procedures also gave positive DNA results with some subclinically infected animals when these only gave suspicious or negative results in the ELISA test. Most clinically and subclinically infected animals were detected with milk PCR. CONCLUSION: Since there may well be a haematological phase in paratuberculosis, nested PCR testing of blood and milk samples shows potential to detect animals subclinically infected with M a paratuberculosis. More subclinically infected animals need to be tested and confirmed infected before estimates of sensitivity and specificity can be made. PMID- 15359967 TI - Intrauterine and transmammary transmission of Mycobacterium avium subsp paratuberculosis in sheep. AB - OBJECTIVE: To investigate intrauterine infection of foetuses with Mycobacterium avium subsp paratuberculosis and the presence of infection in mammary secretions of sheep. DESIGN: A study of 142 late-pregnant ewes and their foetuses from two heavily infected flocks. PROCEDURE: Infection of ewes was determined at necropsy by histopathology and culture of tissues and mammary secretions. Antemortem tests (clinical assessment, faecal culture and serology) were also applied. Foetuses from 59 infected ewes and 47 apparently uninfected ewes were examined by culture and histopathology. RESULTS: Five of five ewes with clinical ovine Johne's disease had infected foetuses. Only one of 54 subclinically affected ewes, and none of 47 uninfected ewes had an infected foetus. M a paratuberculosis was cultured from mammary secretions or mammary glands of only two of 76 ewes, both of which were clinical cases and had infected foetuses. CONCLUSION: Although intrauterine or transmammary transmission of Mycobacterium avium subsp paratuberculosis may occur frequently in clinically affected sheep, these are less common in subclinically infected ewes. Therefore these modes of transmission are unlikely to compromise existing control programs for ovine Johne's disease on most farms, especially if programs include the immediate culling of clinically affected sheep. PMID- 15359968 TI - An infectious myocarditis syndrome affecting late-term and neonatal piglets. PMID- 15359969 TI - Creating a national, transdisciplinary community of clinicians who serve the underserved. PMID- 15359970 TI - Reducing unintentional injuries on the nation's highways: research and program policy to increase seat belt use. AB - Death, disability, and injury from motor vehicle accidents constitute a public health crisis. The goal of this paper is to describe how Meharry Medical College's Center for Community Based Research plans to address this problem. A model of how high-risk groups are influenced to engage in behaviors that increase risk for traffic crashes is articulated. Five strategies for reducing risk for motor vehicle morbidity and mortality are identified: 1) influencing the individual at the point of decision; 2) mobilizing communities and coalitions to support individual and systems changes; 3) modifying environmental factors to modify behaviors; 4) changing laws and public policy; and 5) working towards the elimination of underlying causes. The Center for Community Based Research's promotion of seat belt use, based on each of these five strategies, is described. Addressing the public health crisis resulting from death and injury on the nation's roads and the excess risk faced by minority groups in this country will require the coordinated efforts of many groups. This work must be driven by research, the outcome of which will be a reduction in preventable injury, disability and premature death. PMID- 15359971 TI - Collaborative/Participatory research. AB - Collaborative (or, participatory) research involves a working relationship between at least one academic institution's research unit and one community-based organization. The community-based organizations of interest are those from and representing underserved communities. Barriers to collaboration, approaches to overcoming such barriers, and principles for the maintenance of good collaborative research relations are given. Representatives of the underserved and academic research units tend to interact in a collaborative/participatory relationship by means of formal committees. How the degrees of power of the underserved on such committees might be understood is discussed in terms of a model from Arnstein's 1971 article and the value of participation for the underserved is discussed. PMID- 15359972 TI - Churning in Medicaid managed care and its effect on accountability. AB - There is concern that churning in Medicaid excludes children from the accountability system for managed care because they may not meet the one-year continuous enrollment requirement. This study explores the effect of churning in measuring childhood immunization coverage rates under the current accountability system. Data were collected from administrative databases at the Centers for Medicaid and Medicare Services and 12 states with high Medicaid managed care penetration. On average in the 12 states only 39% of the children enrolled in one specific managed care plan met the continuous enrollment requirement. However, Centers for Medicaid and Medicare Services data showed that 78% of children were enrolled in Medicaid (but not the same plan) continuously for 12 months. Both plan-specific rates and overall Medicaid rates varied greatly across the states. Policies that result in churning mean that many vulnerable children fall outside of the accountability structure intended to assure that they receive necessary services. PMID- 15359973 TI - Free clinics helping to patch the safety net. AB - For the uninsured and underinsured with few funds, there are a limited number of health care options. To assist in filling this hole in the safety net, hundreds of free or volunteer-based clinics have been established across the country. Although these clinics have existed for years, little data on them exist. In 1999, a mail survey was sent to free clinics in seven Midwestern states. Findings from this survey show that, in a single year, these 106 clinics provided medical, dental, and pharmaceutical services to over 200,000 patients, suggesting that free clinics nationwide are caring for a substantial number of our nation's uninsured. The survey paints a picture of free clinics, the populations they serve, and the services they provide. Given the lack of options for the uninsured and underinsured, free clinics deserve continued recognition, support, and assistance from policy makers, health care providers, and the philanthropic community. PMID- 15359974 TI - Full disclosure of financial costs and options to patients: the roles of race, age, health insurance, and usual source for care. AB - The objective was to identify factors associated with financial discussions and financial disclosure of medical costs within a low-income urban community. The method used was a cross-sectional community-based survey in Allegheny County, Pennsylvania. The survey was conducted door-to-door and at area food pantries. Two hundred and twenty six adults were interviewed. Overall, 76.1% reported having a usual source for care and 73.0% had health insurance. Thirty nine and four tenths percent reported having been asked about their ability to pay for health services; this was more common among African Americans (OR 5.2; 95% CI 1.73-15.84), those with no health insurance (OR 4.3; 95% CI 1.01-17.89), and those less than 45 years old (OR:2.9; 95% CI 1.03-8.28). Only 10.6% reported being told how much a health visit would cost. Overall, 30.1% reported their provider made payment allowances for medical bills, with white respondents 2.5 times more likely and those persons identifying an ambulatory site for care 2.6 times more likely to report this. Overall, 30.5% reported being referred to a collection agency for unpaid medical bills; this was 2.4 times more common among those individuals identifying a non-ambulatory usual site for care. Significant race and socio-economic disparities exist in discussions about and access to financial resources to pay for medical care. Expanding the availability of financial assistance is critical to improving access to health care. PMID- 15359975 TI - Do medical out-of-pocket expenses thrust families into poverty? AB - This paper estimates the impact of medical out-of-pocket expenses on families' well-being using the Survey of Income and Program Participation. Medical out-of pocket expenses include the out-of-pocket costs from medical services and the family's share of health insurance premiums. Demographic characteristics, insurance status, and medical usage of the family are analyzed to determine which characteristics are most likely to impoverish a family. Families impoverished because of medical out-of-pocket expenses are far more likely to have older heads of the family, at least one family member in poor health, or some adults without health insurance. Families without at least one person who worked full time for the entire year were also likely to be impoverished. However, children in the family had little effect on the probability that the family became impoverished. This odd result is probably due to the high correlation between parental health insurance coverage and the health insurance coverage of their children. PMID- 15359976 TI - Reducing unintentional injuries on the nation's highways: a literature review. AB - Death and injury on the nation's highways is a public health crisis, especially for youth and members of selected minority groups. The objective of this paper is to review the literature on behavioral and environmental factors that increase risk for traffic morbidity and mortality in populations at high risk. Each of the following is a risky traffic-related behavior: not wearing seat belts, not using child safety seats, not wearing bicycle or motorcycle helmets, driving after drinking, driving while fatigued or distracted, speeding, running red lights, and aggressive driving. Environmental factors that modify risk include urban sprawl, highway design, public policy, racism and economic inequality. High risk groups include youths, males, pickup truck drivers, urban dwellers, the elderly, African Americans, American Indians, and Alaska Natives. A comprehensive approach must be developed for reducing traffic-related risk of death and injury, especially in high risk populations. PMID- 15359977 TI - Prevalence and detection of urinary incontinence among older Medicaid recipients. AB - Low-income older adults have higher rates of many medical disorders than those with higher income, but rates of urinary incontinence have not been examined in this population. A random sample of older Medicaid recipients was interviewed (n=910) and medical records examined for the subset with urinary incontinence (n=236). Nursing home residents were randomly selected from Medicaid enrollment files (n=480). Forty-two percent of community residents reported urinary incontinence, with higher rates among women, older respondents, and whites. The medical records for only 22% of community-dwellers contained a diagnosis of urinary incontinence, compared with 77% for nursing home residents. Type of urinary incontinence was specified for 65% of diagnosed community dwellers and 7% of diagnosed nursing home residents. Urinary incontinence rates are high among Medicaid recipients compared with estimates from general population studies, but detection rates are lower for community-dwellers. Physicians may need to do more among low-income older adults in order to detect urinary incontinence. PMID- 15359978 TI - Demographics of disenrollment from SCHIP: evidence from NJ KidCare. AB - The State Children's Health Insurance Program (SCHIP) provides health insurance coverage for children in low-income families. Although there is evidence of substantial disenrollment from SCHIP, few studies have examined how disenrollment varies by demographic characteristics. This study uses data from administrative records of all 41,881 children enrolled prior to April 2000 in NJ KidCare (New Jersey's SCHIP) separate state plans for families with incomes between 133% and 350% of the Federal Poverty Level. Survival methods were used to analyze disenrollment according to demographic and plan characteristics. Reasons for disenrollment were also studied. Overall, 18.9% of children disenrolled within 12 months of enrollment. Disenrollment was higher among non-Hispanic black children, children aged 1 to 5, and children without siblings in NJ KidCare than among their counterparts. Surprisingly, English speakers had the highest disenrollment rate of all language groups. Children in families with moderate income categories for whom premium contributions were required were 3 times as likely as lower income children to disenroll, principally due to non-payment of premiums. To maximize retention in SCHIP and ensure access to care and continuity of care for low-income children, research is needed concerning why some groups disenroll more quickly. PMID- 15359979 TI - Improving the immunization and health status of children in the Women, Infants, and Children (WIC) Program. AB - Maintaining enrollment in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) and continued exposure of these children to immunization-promoting and nutritional benefits within the program is essential to improve the health status of this vulnerable population. Logistic regression was used to determine characteristics of 2 groups of children: those who dropped out of the program despite being eligible and those who remained in the program but were underimmunized. Of over 20,000 children 19-35 months old, 49% had participated in WIC but only 50% were still enrolled. Factors most strongly associated with dropping out of the program were older age of child; white, black, or American Indian race; living in an urban or suburban area; higher socioeconomic status but still eligible for the program; having only 1 child at home; and having mothers who were unmarried or less than 30 years old (p<0.05). Among current participants, factors most strongly associated with under vaccination included younger age of the child; black or Asian race; moving from another state since birth; mother with less than a high-school education; and having 2 or more children under 18 years old living in the household (p<0.05). Routinely collected child/family information can be used to target outreach and immunization-promoting interventions toward children most likely to drop out of the program or to be underimmunized. PMID- 15359980 TI - [Magnesium as a food supplement]. AB - Magnesium is a Cofactor in all enzymes transfering phosphate. It is therefore necessary for the energymetabolism and for many synthesis, i.e. DNA and RNA. Mg is one of the most widespread elements of the earthcrust and is found in many nutritients. Nevertheless the daily intake in the western countries is often under the recommended level, due to the western life stile. Regular alcohol intake lessens the resorption and regular sport enhances the excretion via the sweat. Especially in elderly people this can lead to a lack of Magnesium. This may promote muscular cramps and weakness, cardiac arrhythmias, hypertension or bronchial hyperreactivity. It seems to be reasonable to recommend liberally a oral supplementation with magnesium. PMID- 15359981 TI - [Cancer in the elderly]. AB - The increase in cancer incidence with increasing age is becoming more obvious and more important as the average age of population increases. Currently over 50% of all cancers occur in people who are 65 and older. The clinical behaviour of common malignant diseases may change with age because of intrinsic variation of the neoplastic cells and the ability of the tumor host to support neoplastic growth. Therapeutic decisions should be based on an estimation of patient's life expectancy, functional status, comorbidity, polypharmacy, socioeconomic conditions, nutrition and the presence of geriatric syndromes. This review should give some answers to the many existing questions about cancer in the elderly. PMID- 15359982 TI - Sleep apnea syndrome and its complications. AB - In this article we summarize the available information regarding the epidemiology, the pathophysiology as well as the risk factors and complications of the sleep apnea syndrome (SAS). Central, obstructive and mixed forms of SAS are known, however, the obstructive form is (resulting from the actual high prevalence of obesity) definitely the most frequent. Latest years of experimental and clinical research have pointed towards the clinical importance of this sleep related breathing disorder. High prevalence in the population and especially the cardiovascular complications (e. g. systemic and pulmonary hypertension, atherosclerosis, arrhythmias) have contributed to the recent increase in knowledge about SAS. Nevertheless, there are numerous unsolved problems and unanswered questions in the pathophysiology of SAS. Future studies should, thus, provide us with more information and shed light on regarding the hidden mysteries of SAS. PMID- 15359983 TI - Methods of GFR determination--creatinine clearance after cimetidine administration in clinical practice. AB - Determination of clearance of endogenous creatinine using its plasma and urinary concentration (standard clearance) or Cockroft and Gault equation (estimated clearance) is commonly performed for assessment of glomerular filtration rate. Although evaluation of renal function in this way is useful, it is biased with an error resulting from secretion of creatinine in tubules. This error can be reduced by determining the clearance after administration of cimetidine. This study enrolled 40 patients who were divided into two groups--the first group consisted of 12 healthy subjects and the second group joined healthy subjects and patients with chronic renal failure. The second group was divided into three subgroups in relation to creatinine serum concentration (intervals: creatinine < 1 mg/dL - 9 subjects, creatinine 1-2 mg/dL - 7 subjects, creatinine > 2 mg/dL - 10 subjects). Estimated and standard creatinine clearance, clearance after cimetidine administration and inulin clearance were assessed. Analysis of mean percentage differences between clearance value after cimetidine administration and estimated or standard clearance shows increasing contribution of tubular creatinine secretion with increase in plasma creatinine concentration. This study suggests that determination of creatinine clearance after cimetidine administration can be used in precise evaluation of renal function. PMID- 15359984 TI - Transient hypoglycemic abducens palsy. AB - Since 1928, among the thousands of patients treated for insulinoma, only 32 cases with peripheral neuropathy have been reported. None of these described an affection of the cranial nerves. We present a 56 old woman, who suffered from chronic hyperinsulinism due to an insulinoma. For ten years, the patient has developed progressively marked hypoglycemic attacks of up to 20 mg/dl. Recently we have observed the development of a paresis of the right abducens nerve lasting for 6 weeks. PMID- 15359985 TI - Recurrent malignant schwannoma of the small bowel. AB - Primary malignant schwannoma of the small bowel is an extremely rare disease. Only 24 malignant schwannomas of the small bowel have been reported in the medical English literature. We report a primary malignant schwannoma of the distal ileum in a 53-year-old woman, who was admitted to the hospital because of abdominal pain. CT scan revealed a semisolid mass in the small intestine of the serosa. The tumour measuring 14 x 13 x 8 cm and 100 cm of the distal ileum were resected. We observed a recurrence after seventeen months and the recurrent tumour measured 6 x 2.5 x 1 cm was excised. PMID- 15359986 TI - Chromosome 13 deletion/hypodiploidy and prognosis in multiple myeloma patients. AB - The application of high-dose treatment with autologous stem cell transplant(s) has improved survival, when compared to standard treatment, in patients with multiple myeloma. However, this benefit is mostly enjoyed by specific patient subgroups characterized by the absence of high-risk disease features. High-risk features are, first and foremost, the detection of unfavorable cytogenetic abnormalities (chromosome 13 deletion, hypodiploidy and myelodysplastic-type abnormalities in an otherwise typical myeloma karyotype) prior to treatment; elevated serum lactate dehydrogenase and C-reactive protein levels at diagnosis and high beta-2 microglobulin levels prior to transplant also convey poor prognosis, although they account for less variability of the observed outcome than the cytogenetic abnormalities. While high-dose treatment with autologous stem cell transplant(s) can cure a sizable minority of patients with low-risk disease features and significantly prolongs survival in others with similar characteristics, patients with high-risk features are virtually incurable and their survival benefit is much less pronounced. As the tremendous clinical variability of myeloma can now be traced to its underlying genetic abnormalities, routine cytogenetic analysis at diagnosis and relapse are absolutely indicated. Based on this stratification, high-risk patients are excellent candidates for novel therapeutic approaches, such as planned non-myeloablative allogeneic transplants following an autologous transplant. PMID- 15359987 TI - Immune reconstitution following stem cell transplantation. AB - The period of immune deficiency following stem cell transplantation (SCT) results in significant morbidity and mortality. Whilst supportive therapies have partially improved the outcome of infective episodes, disease relapse remains a considerable obstacle to improvement in overall outcomes. An increased understanding of the importance of the immune system in preventing relapse has derived from studies in the allogeneic setting. Increasing awareness of autologous anti-tumor responses has also focused interest on enhancing such activity. Successful application of some of these newer therapies, such as tumor vaccination approaches, may depend critically on reconstitution of functional immune reactivity. Whilst early recovery of innate immunity (myeloid series and natural killer (NK) cells) results in reconstitution of protective immunity against many bacterial pathogens, both the absolute levels and function of T and B lymphocytes remain abnormal for many months or years. Incorporation of T-cell depletion, choice of graft type (both donor and source), development of graft-vs. host disease and level of residual thymic activity can all influence aspects of the reconstitution process. Advances in immunological monitoring are providing new insights, particularly into the recovery of specific T-cell subsets. This review focuses mainly on recent advances in the understanding of immune reconstitution in the allogeneic setting. PMID- 15359988 TI - Cigarette smoking and acute leukemia. AB - Recent cohort and case-control studies have suggested that cigarette smoking may be involved in the etiology of leukemia. Rising trends have been observed for all leukemias when the amount of cigarettes smoked increased. However, the magnitude of the trend was strongest for myeloid leukemia. Although no detailed biological mechanism has been proposed, a causal link is made plausible by evidence of systemic effects of cigarette smoke and the presence in cigarette smoke of chemicals (benzene) and radioactive substances that have been associated with leukemia risk. Cigarette smoking has a deleterious effect on survival in leukemia by shortening complete remission duration and subsequent survival. The data reported in this review are derived from the medical literature and from the experience of the authors. PMID- 15359989 TI - The high rate of bone resorption in multiple myeloma is due to RANK (receptor activator of nuclear factor-kappaB) and RANK Ligand expression. AB - The excessive bone resorption observed in multiple myeloma may be due to the production of several osteoclast-activating factors either by the myeloma cells themselves or by the bone marrow microenvironment. These factors could act primarily via a common final pathway involving the recently-described members of the TNF receptor-ligand family: RANKL (Receptor Activator of NK-kappaB Ligand) and its corresponding RANK receptor that play a crucial role in osteoclast differentiation and activation, and osteoprotegerin (OPG), the physiological inhibitor of RANKL. RANKL expression by stromal cells is increased in myeloma and is associated with a concomitant decrease in OPG expression. This increase in RANKL-OPG ratio correlates with the extent of the myeloma bone disease. The RANKL OPG imbalance could play a decisive role in the lytic bone lesions in myeloma, and this possibility is reinforced by several in-vivo studies that have assessed the effects of administering RANKL inhibitors in murine myeloma models. Treatment with either OPG: Fc or RANK: Fc decreased myeloma osteolysis in these models. RANKL blockade is also currently being evaluated in malignant osteolysis in humans. A therapeutic approach targeting the RANKL-RANK signaling pathway could be of great value, as RANKL inhibitors are potent anti-resorptive agents, affecting both myeloma-induced bone resorption and the tumor burden. PMID- 15359990 TI - Quantitative monitoring of BCR--ABL transcript--suggestion of a simplified approach considering inaccuracy of measurement and calibration. AB - According to standard-protocols, real time quantitative RT-PCR (RQ-PCR) for quantification of BCR-ABL fusion transcripts in CML patients is performed with the construction of a standard curve for each run, each sample is analyzed at least in duplicate and 10-40 ml peripheral blood are processed. This approach is appropriate for a research laboratory, but is not suitable for a routine laboratory setting. We show that the calibration curve based on the common 5 dilution standards (between 10 and 10(6) copies) is strongly influenced by the large variability of the measurements below 100 copies of the gene. In other words, including a standard with 10 copies is a source of error, which cannot be reduced through the construction of a standard curve with each run. Adding additional dilutions between 10 and 100 copies to the standard curve, the variance of the obtained curve is much reduced. As a conclusion, it is unnecessary to construct a calibration curve with each run since only negligible inaccuracy of calibration is added to the inaccuracy of measurement. Running of the samples in duplicate seems unnecessary since the inaccuracy of the method can be correctly estimated. Finally, we propose a standardized collection and isolation of total RNA from only 2.5 ml blood using an integrated system, which allows RNA stabilization for up to 5 days and provides snapshots of BCR-ABL fusion transcripts with higher accuracy than with non-stabilized blood samples. PMID- 15359991 TI - Problems related to resistance to cytarabine in acute myeloid leukemia. AB - First-line chemotherapy treatment in acute-myeloid leukemia patients usually consists of a combination of cytarabine (ara-C) and an anthracycline. These regimens induce complete response (CR) rates in 65-80% of newly diagnosed AML patients. However, clinical outcome is unsatisfactory, as most of the patients who achieve a CR will relapse within 2 years from diagnosis, often with resistant disease and poor response to subsequent therapy. Thus, understanding the factors which contribute to the emergence of chemoresistant leukemic cells is essential to improve outcome in patients suffering from this disease. In this review, we highlight the current knowledge concerning the cellular mechanisms of resistance to ara-C. We also discuss possible strategies that may be used to overcome such resistance. Efforts to increase intracellular levels and DNA incorporation of phosphorylated ara-C using pronucleotides of ara-C are very promising. Ara-C combined with agents modulating apototic responses are expected to provide additional benefit. In the same way that combination chemotherapy has provided curative treatment of AML, a multifactorial approach of ara-C resistance should allow significant progress in the treatment of currently chemoresistant disease. PMID- 15359992 TI - Prognostic factors in Hodgkin's disease. AB - Hodgkin's disease (HD) is a curable tumoral disease. However, there are groups of patients who suffer relapse and the identification of prognostic factors and the adaptation of treatments to individual risk is one the lines of investigation in this disease. A study was performed on 526 patients diagnosed of HD in our hospital between January 1967 and September 2001. An analysis was made of the most important variables in terms of both disease-free and overall survival. Overall survival in this series of patients was 94% at 2 years, 86% at 5 years, 76% at 10 years and 72% at 15 years. Median survival was 249 months. Factors influencing poor prognosis in the overall survival were: male gender (P < 0.0001), lymphocyte depletion (P < 0.0001), stages III and IV (P < 0.0001), B symptoms (P < 0.0001), spleen involvement at diagnosis (P = 0.003), no complete remission after first line treatment (P < 0.0001), and more than 30 years-of-age (P < 0.0001). Disease free survival was 83% at 2 years and 68% at 5 years although without reaching the mean follow-up. The disease free survival study revealed the following risk factors: male gender (P = 0.02), lymphocyte depletion (P < 0.0001), stages III and IV (P < 0.001), B symptoms (P < 0.001), extranodal or splenic involvement (P < 0.05), and no complete remission after first line treatment (P < 0.0001). The result of treatment optimization is that some factors that were considered to indicate a poor prognosis have disappeared, and that others which are useful have appeared and allow us to establish groups with differing risks of relapse and who could be candidates for differentiated treatments. PMID- 15359993 TI - Fludarabine, cyclophosphamide and mitoxantrone for untreated follicular lymphoma: a report from the non-Hodgkin's lymphoma co-operative study group. AB - The aim of the study was to determine the safety and efficacy of the combination of fludarabine (FLU), cyclophosphamide (CY) and mitoxantrone (FLU/CY/MITO) in untreated follicular lymphomas (FL), Sixty patients with newly diagnosed stage II bulky to IV FL, median age 59 years (range 36-70), received FLU/CY/MITO regimen (FLU 25 mg/m2 days 1-3, CY 300 mg/m2 days 1-3, Mito 10 mg/m2 day 1). Patients received antibiotic oral prophylaxis during all treatments, and growth factors (G CSF) when grade III granulocytopenia (WHO) occurred. The overall response rate was 87%: 46 patients achieved complete response (CR) (77%), 6 a partial response (10%) and 8 were non-responders. Fifty patients are surviving with a median observation time of 31 months. The 4-year estimated probability of overall survival and failure-free survival were 78.2% and 45% respectively. Thirty-five patients (58%) are still in CR. Sixty percent of patients experienced grade III IV granulocytopenia. Two patients suffered grade III pulmonary infection and one grade III liver toxicity. In a subset of 46 patients, bcl-2 translocation was positive in bone marrow (BM) and/or peripheral blood (PB) of 36 patients. At the end of treatment, 25 of these patients had CR and 19 (76%) converted to polymerase chain reaction (PCR) negativity. FLU/CY/MITO regimen showed a high level of activity in follicular lymphoma. Toxicity, mainly hematological, was acceptable and the treatment was made feasible by the use of antibiotic prophylaxis and G-CSF. Significant non-hematological toxicities were seen, but no patients died. The conversion of bcl-2 from positive to negative by PCR in BM and/or PB suggests a possible role for this treatment in clearing minimal residual disease and improving patients' outcome. PMID- 15359994 TI - Survival in young patients (less than 40 years) with follicular lymphoma: a population based study by the Scotland and Newcastle Lymphoma Group. AB - We have examined in a population-based observational study the survival of young patients (less than 40 years) with follicular lymphoma (FL) treated conventionally and followed for up to 17 years (minimum 10, median 13 years). Data were derived from the Scotland and Newcastle Lymphoma Group (SNLG) database from 1986. Histology of all available cases was reviewed to ensure that patients met the modern criteria for diagnosis of FL. Of 55 patients identified from the database, 46 were confirmed to have follicular lymphoma. There were 25 males and 21 females, median age 34 years (range 16-39). Thirty-four patients presented with advanced stage disease (Stages III and IV). The majority of patients received initial treatment with chemotherapy, though 7 had surgery (biopsy or splenectomy) alone and 7 radiotherapy alone. All 12 patients with early stage disease showed a complete response (CR) with initial therapy; 6 relapsed and 2 have died (1 of transformation to high grade non-Hodgkin's lymphoma). Overall survival of patients presenting with stage IIIA disease was 68% at 10 years, and 69% for patients in stages IIIB and IV. The SNLG prognostic index for low grade non-Hodgkin's lymphoma was predictive for overall survival. The 71% overall survival in this patient cohort at 10 years provides a baseline for comparison with the results of a more aggressive approach to treatment. PMID- 15359995 TI - Efficacy and safety of fludarabine and cyclophosphamide combined therapy in patients with refractory/recurrent B-cell chronic lymphocytic leukaemia (B-CLL) Polish multicentre study. AB - The aim of this study was to investigate the efficacy of a combination of fludarabine (F) and cyclophosphamide (C) in the treatment of patients with refractory/recurrent B-cell chronic lymphocytic leukaemia (B-CLL). Between November 1999 and December 2001, 63 patients with B-CLL (median age 60 years) received a regimen that consisted of F 25 mg/m2 and C 250 mg/m2, days 1-3, intravenously, every 4 weeks, for a maximum of 6 courses, Response and toxicity were assessed according to current criteria (NCI-WG and WHO). Complete and partial remissions were achieved in 17.5% and 55.6% of patients, respectively; 19% of patients had stable disease and 7.9% of patients showed disease progression. The median follow-up was 16.5 (range 1.5-32) months. The median duration of progression-free survival (PFS) has not been reached among patients treated with FC regimen as second-line therapy. The median PFS was 13 (range 8 26) months in the 19 responding patients treated with FC regimen as third-line therapy. The most frequent side-effects were neutropenia (45%), thrombocytopenia (42%) and infections (57%). We conclude that the combination of fludarabine and cyclophosphamide demonstrated significant efficacy in pretreated, advanced B-CLL patients, with tolerable toxicity. PMID- 15359996 TI - Quantification of CD38 expression in B-cell chronic lymphocytic leukemia (B-CLL): a comparison between antibody binding capacity (ABC) and relative median fluorescence (RMF). AB - We have previously shown that quantification of CD38 expression using microbeads of specific antibody binding capacity (ABC) improves the prognostic value of CD38 expression in B-cell chronic lymphocytic leukemia, particularly for Binet Stage A patients. Quantification of CD38 expression using beads is expensive, time consuming and could be difficult to implement in a routine clinical laboratory. The calculation of relative median fluorescence (RMF) using the median fluorescence intensities of the test and control samples, is even more simply and cheaply obtained by flow cytometry and could be used as an alternative way of quantifying antigen expression. The present study demonstrates that RMF is an effective prognostic indicator in B-CLL that correlates closely with ABC in predicting disease-specific survival and time to progression for all patients. RMF predicted overall survival and time to progression in all patients (P < 0.0001 for both), in Binet Stage A patients (P < 0.0001 for both) and in Stage A patients under 60 years (P = 0.0299 and P = 0.0143, respectively). ABC predicted overall survival and time to progression in all patients (P < 0.0001 for both) in Stage A patients (P = 0.0024 and P < 0.0001, respectively) and in Stage A patients under 60 (P = 0.0379 and P = 0.0032, respectively). RMF is more effective than percentage CD38 positivity > 30% or > 20% in predicting disease specific survival in Stage A patients of all ages (CD38 < > 30%: P = 0.0853, CD38 < > 20%: P = 0.0894) and in those under 60 years old (CD38 < > 30%: P = 0.5438, CD38 < > 20%: P = 0.2872). Also, RMF is more effective in predicting time to progression of Binet Stage A patients less than 60 years (P = 0.0143), while percentage CD38 positivity of 30%, 20% or 7% did not achieve statistical significance (P = 0.1103, = 0.0547, = 0.3399, respectively). We suggest that CD38 RMF could be used clinically as an alternative to ABC to identify patients with B CLL that are likely to progress and require early treatment. PMID- 15359997 TI - Hematopoietic stem cell transplantation in adult acute lymphoblastic leukemia: a single-centre analysis. AB - The role of autologous or allogeneic hematopoietic stem cell transplantation (HSCT) in ALL is controversial because of its adverse risk/benefit ratio, and the main policy is to reserve it for high-risk patients. In our Institution, between 1984 and 2002, 40 patients received an allogeneic HSCT and 39 underwent autografting. The conditioning regimen included HD-Ara-C, HD-CTX and 10 Gy fractionated TBI. After allogeneic SCT in first CR, four patients relapsed, leading to a 10-year EFS chance of 78.3%; of the other patients, 5 are still in CR. After autografting in first CR, there was an early death, one secondary AML, one death in CR and six relapses, leading to a 10-year EFS chance of 44.4%; of the other patients, 6 are still in CR. Even considering the limited number of patients and the slow accrual rate, selection bias cannot be considered a sufficient explanation for the favorable outcome of allografting in first CR as the majority of the patients had adverse prognostic factors. It cannot be claimed that allogeneic SCT was performed in patients already cured, as the autografted patients had a notably worse outcome, and a 10-year EFS chance of about 80% is an uncommon finding even in standard-risk ALL patients. It might be inferred that the timing of SCT as late intensification, in addition to a rather aggressive conditioning regimen, helped to minimize the leukemic burden, thus favoring the expression of a GVL effect. Conversely, the results in more advanced disease phases are discouraging, due to poor quality CRs and inefficacy of GVL in managing large residual disease. PMID- 15359998 TI - Constitutive NF-kappab/Rel activation in philadelphia chromosome positive (Ph+) acute lymphoblastic leukemia (ALL). AB - The Bcr-Abl translocation t(9;22)(q34;q11 ) defines a subgroup of ALL patients with a dismal prognosis despite the introduction of intensified induction and consolidation regimen. Although Bcr-Abl induced NF-kappaB/Rel activation has previously been shown, the role of NF-kappaB/Rel in Ph+ leukemia is unclear. Using DNA binding assays, we demonstrate constitutive NF-kappaB/Rel activity in nuclear extracts from Ph+ ALL blasts, whereas Ph- primary blast cells and B precursor cell lines lack NF-kappaB/Rel activity. NF-kappaB/Rel activity was shown in the ela2 and the b2a2 subtypes. Constitutive NF-kappaB/Rel activity in Ph+ blasts is not due to elevated endogenous IkappaB kinase (IKK) activity as shown by immune complex kinase assays. Since NF-kappaB/Rel is a transcriptional regulator of inhibitors of apoptosis we examined the expression of anti-apoptotic genes known to be induced by NF-kappaB/Rel by real time PCR analysis. We found no induction of TRAFI, TRAF2, cIAPI, cIAP2, XIAP, A20 or Bfl/Al in Ph+ ALL samples as compared to Ph-negative ALL controls. In summary, constitutive NF-kappaB/Rel activation independent of endogenous IKK activation may be a common finding in Ph+ ALL. However, targets of NF-kappaB/Rel mediated transcriptional regulation in this disease remain to be identified. PMID- 15359999 TI - Bone marrow microvessel density (MVD) in adult acute myeloid leukemia (AML): therapy induced changes and effects on survival. AB - Based on the strong evidence in favor of an increase in microvessel density (MVD) in hematological malignancies, we evaluated VEGF immunoreactivity and MVD measurement in bone marrow biopsies of 36 AML patients at diagnosis and following therapy. MVD assessment was based on CD31, CD34 expressing vessels. The values were calculated for only one marker if the other vascular marker was positive on blasts, otherwise both markers were used. VEGF immunoreactivity was also scored. Comparison of MVD values of 36 AML patients with 18 non-malignant controls showed a significantly higher MVD in AML (CD31: P = 0.004, CD34: P < 0.001), which is independent of other variables such as cellularity or blast percentage. Following induction chemotherapy, the responders showed a significant decrease in blast counts (P < 0.001), cellularity (P = 0.001) and MVD (P = 0.050) quantification with CD31. Higher baseline MVD (CD34) values were associated with shorter overall survival (P = 0.0027). These results are encouraging for inclusion of MVD enumeration in bone marrow examinations of AML patients at diagnosis as an additional prognostic factor. PMID- 15360000 TI - Allogeneic hematopoietic stem cell transplantation with non-myeloablative conditioning in patients with acute myelogenous leukemia eligible for conventional allografting: a prospective study. AB - Using a non-myeloablative stem cell trasplantation (NST) program, 25 allografts were prospectively given to 24 patients with acute myelogenous leukemia (AML) eligible for conventional allografting; 2 individuals had secondary forms of AML. The median age of the patients was 35 years, with a range of 12 to 56. All patients engrafted; median time to achieve an absolute neutrophil count > 0.5 x 10(9)/1 was 12 days (range 0-26), whereas the median time to a platelet count > 20 x 10(9)/1 was 13 days (range 0-26). Patients developed mixed chimerism 15 to 100 (median 30) days after the allograft. The follow-up periods range between 33 and 2670 days (median 450). The median post-transplant overall survival of the patients has not been reached and is above 89 months, whereas the 683 days both overall and progression-free survival is 66%. In 14 grafts (56%) acute GVHD ensued; in 12 cases grades I-II and in 2 cases grade IV which was fatal in both. In 9/19 patients (47%) limited chronic GVHD developed. In 22 cases (88%), the procedure could be completed fully on an outpatient basis. The 100-day and the transplant-related mortality were both 8%. NST appears to be an effective additional therapeutic option for patients with AML in remission and a matched donor available. PMID- 15360001 TI - Abnormalities of chromosomes 1p34-36, 4p16, 4q35, 9q11-32 and +7 represent novel recurrent cytogenetic rearrangements in chronic lymphocytic leukemia. AB - Karyotypes were studied in over 250 cases of CLL seen at our Institution and 12 cases with a previously undescribed chromosome abnormality were identified. Cytogenetic and clinicobiological features in these patients are described. Fluorescence in situ hybridization using probes for the detection of +12 and deletions of 13q14, 17p13, and 11q22-23 was performed. Hematologic and clinical data were reviewed and a review of the literature was performed. Twelve patients were found carrying the following aberrations in the stemline: abnormalities at 1p34 (n = 2), 4p16 (n = 2), 4q35 (n = 2), 9q11-32 (n = 4) and +7 (n = 2). Trisomy 12 was found in 3 cases, whereas no case carried 13q-, 11q-, 17p-. Our data showed that (i) aberrations involving 1p34 and 4p16 as isolated chromosome anomalies were preferentially associated with early stage disease; (ii) 4q35 anomalies were associated with a relatively aggressive disease, atypical morphology and with monoclonal gammopathy; (iii) rearrangements of 9q were characterized by atypical morphology and aggressive disease with splenic involvement; (iv) +7 be may associated with +12. 1p34-36; 4p16; 4q35; 9q and chromosome 7 represent novel recurrent rearranged sites in CLL, with a 0.5-3% incidence. Transformation in these patients seemingly occured through a cytogenetic route not involving the classical CLL-associated chromosome regions. These chromosome rearrangements may be associated with peculiar hematologic features. PMID- 15360002 TI - P73 status in B-cell chronic lymphocytic leukaemia. AB - B-cell chronic lymphocytic leukemia is a heterogenous disease with disturbed apoptosis in which the precise molecular defects leading to this pathogenesis are still unclear. The p73 gene (a p53 homologue) encodes 2 proteins with opposing functions. TAp73 induces cell cycle arrest and apoptosis, whilst the oncogenic deltaNp73 inhibits both TAp73 and p53 induced apoptosis. Microsatellite analysis was performed to investigate the p73 gene locus in B-CLL. Moreover, we investigated the expression of the TAp73 and deltaNp73 variant by measuring the mRNA transcripts in 51 B-CLL patients by real-time RT-PCR. And in addition, protein expression was analyzed by Western blotting technique in 20 B-CLL patients. There was no evidence of clonal loss of heterozygosity at 1p36, the p73 gene locus in B-CLL patients. The real time RT-PCR analysis showed that the expression of both p73 gene variants was much higher in leukemic cells compared to controls. In 17/20 (85%) patients deltaNp73 and TAp73 protein were present. The observed increase of expression of the antiapoptotic deltaNp73 variant in neoplastic cells may lead to a functional p53 inactivation. This mechanism might be relevant in malignancies with an intact p53 gene but disturbed apoptosis mechanisms such as in B-CLL. PMID- 15360003 TI - Evaluation of temozolomide in patients with myelodysplastic syndrome. AB - In our previous phase I study of temozolomide in patients with acute leukemia, temozolomide was well tolerated and demonstrated significant anti-leukemic activity. The maximum tolerated dose was 200 mg/m2/d for 7 days, repeated approximately every 5-6 weeks. In the current study, we evaluated the same dose of temozolomide in patients with myelodysplastic syndrome. Fourteen patients received 19 courses of temozolomide. The median age was 71 years. In this study, treatment was poorly tolerated with patients requiring admission in 9 of 19 courses. Toxicity included worsening cytopenias, neutropenic fever, and exacerbation of cardiac disease, the latter due to worsening anemia. An unusual finding was the development of leukocytoclastic vasculitis in 4 patients. There were no formal responses to therapy. The current schedule of temozolomide is not efficacious in patients with myelodysplastic syndrome. PMID- 15360004 TI - Expression of HOXA genes in patients with multiple myeloma. AB - Multiple Myeloma (MM) is an incurable B-cell malignancy characterized by uncontrolled growth of plasma cells (PCs) in the bone marrow. The pathogenesis of MM is complex and still not fully understood. The HOX genes encode a family of homeodomain containing transcription factors which are crucial for embryonic development and differentiation. The HOX genes are also involved in hematopoiesis and have been shown to be dysregulated in leukemia suggesting a role in leukemogenesis. We hypothesized that expression of the HOX genes might also be of importance in MM. We screened FACS-sorted malignant PCs from a panel of 32 MM patients for the expression of HOXA 2, 3, 4, 7, 9, 10, and 11 genes by RT-PCR assays specific for each gene. We found that 9.4% (3/32) of the MM patients expressed the tested HOX genes in their PCs suggesting that HOXA genes are frequently dysregulated and might have an oncogenic potential in MM. PMID- 15360005 TI - Heterogeneity of response to imatinib-mesylate (glivec) in patients with hypereosinophilic syndrome: implications for dosing and pathogenesis. AB - Four cases of hypereosinophilic syndrome (HES) treated with the tyrosine-kinase inhibitor imatinib-mesylate are reported. The drug was effective in three patients, but a prolonged clinical and hematological remission was obtained only in one patient, due to appearance of resistance or poor tolerability in the other cases. The dose of imatinib necessary to achieve a response ranged from 100 to 600 mg/d. One patient with evidence of a clonal T-cell population did not respond at all. We confirm the efficacy of imatinib in HES, but we also underline that type and duration of response may be variable. This could be due to different pathogenetic mechanisms of the disease in single patients. PMID- 15360006 TI - Defective immune function of primary effusion lymphoma cells is associated with distinct KSHV gene expression profiles. AB - Primary effusion lymphomas (PEL) are uniformly infected with Kaposi's sarcoma associated herpesvirus (KSHV), and thus likely present both tumor and viral antigens to the immune system. In order to grow unrestricted and cause disease, multiple immune evasion strategies may be utilized by PEL to evade immune surveillance. Using six well-established PEL cell lines and comparing these to Epstein-Barr virus-transformed B cell lines and peripheral blood B cells, significant differences were found in the surface expression of molecules involved in antigen presentation, T cell activation and cell-cell adhesion. Significantly reduced stimulation of cytotoxic T lymphocytes, lowered sensitivity to natural killer cell-mediated lysis and impaired function as antigen presenting cells in mixed leukocyte reactions were found for three PEL cell lines with particularly low CD54, CD58 and CD81 expression. Comparative microarray analysis demonstrated specific patterns of KSHV-encoded gene expression that were associated with the different immune functions of these cell lines. Thus, the present data suggest that distinct patterns of KSHV gene expression may be associated with particular phenotypic and functional characteristics of PEL cells, which may influence PEL pathogenesis. PMID- 15360007 TI - Coexpression of CD40 and CD40L on B lymphoma and carcinoma cells: an autocrine anti-apoptotic role. AB - To evaluate a possible autocrine role of CD40L, the expression and functional activity of CD40L on NHL and breast carcinoma cell lines were investigated. Using flow cytometry, CD40 was consistently detectable at the surface of all 5 NHL cell lines tested. CD40L expression was detectable at the surface of DAUDI (54%, MFI 47) and BJAB (12%, MFI 32) cell lines, and marginally on the RAJI cell line (7%, MFI 30), while 4 of 5 NHL cell lines (DAUDI, RAJI, BJAB, BL70) had detectable CD40L mRNA. CD40 was expressed on T47D and BT20 breast carcinoma cell lines while CD40L was detectable on T47D (93%, MFI 137) only. Both BT20 and T47D had detectable CD40 mRNA, while CD40L mRNA was detectable only in the T47D cell line. CD40, but not CD40L, was detectable on 6 renal, 1 prostatic and 1 colon carcinoma cell lines. CD40L expressed on tumor cells was functional, as shown by its capacity to decrease drug-induced apoptosis on CD40 expressing NHL and breast carcinoma cell lines, while irradiated CD40L negative cell line (BT20) had no effect. Blocking CD40L antibody abrogated the protective effect of irradiated CD40L positive T47D cell line against drug-induced apoptosis on BL70 cell line, confirming that CD40L is functional in the DAUDI and T47D cell lines. Importantly, blocking CD40L antibody increased drug-induced apoptosis in CD40L positive cell lines but had no effect on the CD40L negative cell lines. CD40L is expressed on CD40 positive B NHL and breast carcinoma cell lines and induces an autocrine antiapoptotic signal when cells are exposed to cytotoxic agents. PMID- 15360008 TI - In vitro proliferation and expansion of hematopoietic progenitors from patients with diffuse large B-cell lymphoma before and after chemotherapy. AB - We have previously demonstrated that when cultured in Dexter-type Long-Term Marrow Cultures (LTMC), hematopoietic progenitor cells (HPC) from patients with Diffuse Large B-Cell Lymphoma (DLBCL) show a defective proliferation, as compared to HPC from normal marrow. In that study it was also demonstrated that functional alterations were present in the hematopoietic microenvironment developed in culture; thus, it was not clear whether such a defective proliferation in vitro was due to an intrinsic defect in the HPC compartment of DLBCL patients, or to an altered microenvironment, or both. In order to address this question, in the present study we have assessed the proliferation and expansion potentials of HPC present in bone marrow from patients with DLBCL, in cytokine-supplemented liquid cultures initiated with a cell population enriched for CD34+ Lin- cells, in the absence of stromal cells and in the presence of reduced numbers of accessory cells. Our results demonstrated that bone marrow-derived HPC from patients with DLBCL, both before and right after chemotherapy, possessed reduced proliferation and expansion potentials in vitro, as compared to their normal counterparts. Interestingly, in patients analyzed 18 months after treatment the proliferation and expansion levels were similar to those of normal HPC, indicating a complete restoration of the hematopoietic function. Although the reason for these observations is not clear, our results suggest the possibility that primitive CD34+ progenitor cells present in bone marrow, which show deficient proliferation and expansion potentials in vitro, are involved in the origin/progression of DLBCL. PMID- 15360009 TI - Establishment and characterization of a new mantle cell lymphoma cell line M-1. AB - A new mantle cell lymphoma cell line, M-1, was established from peripheral blood mononuclear cells of a patient with a diagnosis of blastoid variant of mantle cell lymphoma in leukemic phase. This cell line showed cell surface antigens identical to the original tumor and demonstrated the profile of a mature B-cell phenotype typical of mantle cell lymphoma: positive for CD5, CD19, CD20, sIgM and FMC7, and negative for CD3, CD10 and CD23. Cytogenetically, the M-1 cell line showed chromosomal alterations similar to the initial clinical specimen, among which a translocation t(11;14) (q13;q32) resulting in the overexpression of cyclin D1 as well as additional abnormalities involving chromosomes 3, 9 and 10. This cell line was used as a model to investigate the activity of the three drugs doxorubicin, cyclophosphamide and vincristine, commonly used in the treatment of mantle cell lymphoma patients. The effect of the drugs was evaluated by a 24 h cytotoxicity test and a 7-days anti-proliferation test using a microculture tetrazolium-based assay (MTT). Both assays indicated a higher sensitivity of the cell line to vincristine when compared to doxorubicin and cyclophosphamide. The characterization of a new mantle cell lymphoma cell line is a unique tool for studying the biology of this subtype of lymphoma for which only a few cell lines have been established. PMID- 15360010 TI - Richter's syndrome in the brain first manifested as an ischaemic stroke. AB - Isolated central nervous system involvement in Richter's syndrome (RS) is extremely rare and only 6 such cases have been described, so far. We report a 60 year-old woman with B-cell chronic lymphocytic leukemia (B-CLL) heavily pretreated with cladribine based regimens and rituximab in whom RS in the brain was first manifested as a stroke. Initial cranial computed tomography (CT) revealed a hypodense area in the right parietal lobe showing no contrast enhancement. The follow-up CT done after 2 months showed an irregular, slightly hyperdense tumor surrounded by oedema with mass effect and midline shift. However, cerebrospinal fluid (CSF) examinations revealed no pathological changes. Neurosurgical operation was performed and the diagnosis of diffuse large B-cell lymphoma (DLBCL) has been established on the basis of histological and immunological investigation of the tumor. The pattern of immunoglobulin heavy chain (IgH) gene rearrangement in the patients' bone marrow aspirate and brain tumor was identical and suggested that both tumors originated from the same B cell progenitors. The patient was then treated with brain irradiation (2000 cGy) and complete remission as assessed by MRI was achieved. Significant neurological improvement was observed and no clinical progression was stated 3 months after radiotherapy. PMID- 15360011 TI - Aggressive natural killer cell leukemia/lymphoma: case report, use of telesynergy and review of the literature. AB - Natural killer cell malignancies, although increasingly recognized, remain rare tumors within the USA and Europe. They are somewhat more common in Asia, and have been best characterized within this population. We present a case of a young Caucasian woman who presented acutely with an aggressive natural killer cell leukemia/lymphoma. Use of Telesynergy technology enabled a transatlantic telemedicine conference with colleagues in a center of expertise. Unfortunately the patient was ultimately refractory to both conventional chemotherapy and Campath-1H and her disease course was fulminant, as has been described previously in this condition. We review the possible therapeutic options for this extremely aggressive malignancy and briefly discuss our center's experience of telemedicine technology. PMID- 15360012 TI - Somatostatin receptor scintigraphy in MALT lymphoma of the lacrimal gland treated with rituximab. AB - Non-gastric stage I extranodal marginal zone lymphomas of MALT are usually treated with local therapy. However, distant relapses in other extranodal sites are not uncommon suggesting under-staging of these patients using conventional image studies. Positron emission tomography (PET) scans are frequently negative in this subgroup of patients with lymphoma and therefore do not significantly contribute to the staging process. We present a case of lacrimal gland MALT lymphoma demonstrating the usefulness of somatostatin receptor scintigraphy for initial staging and evaluation of response to treatment. PMID- 15360013 TI - Simultaneous appearance of trisomy 8 and trisomy 12 in different cell populations in a patient with untreated B-cell chronic lymphocytic leukemia and myelodysplasia. AB - The co-existence of spontaneously arising myeloid and lymphoid malignancies in the same patient is rare, and is thought to be mainly due to chance. We describe a patient presenting simultaneously with chronic lymphocytic leukemia (CLL) and myelodysplasia (MDS). Histological, flow cytometric, chromosomal and fluorescent in situ hybridization (FISH) studies show that both cell populations possess different sets of markers consistent with the myeloid and lymphoid differentiation pathways. The question of whether these arose from a single or two separate progenitor cells is explored. PMID- 15360014 TI - Granulocytic sarcoma of the ovotestis: an association of myelodysplastic syndrome and hermaphroditism. AB - Granulocytic sarcomas are extramedullary tumors (EMD) of malignant myeloid precursor cells. EMD or granulocytic sarcoma of ovary is rare disease. A 15-year old girl had complaints of abdominal pain and weight loss for 3 months. On physical examination, there were hepatosplenomegaly and a painless mass under the umbilicus. Breast development was grade II. There was no clitoris hypertrophy. Her labia majora were separate and vagina hypoplastic. Hemoglobin level was 9.3 g/dl, white blood cells count 2.8 x 10(6)/1, platelet count 31.6 x 10(9)/1. There were dysplastic features in the blood and bone marrow cells. There were 10 and 22% blasts in the peripheral blood smear and bone marrow, respectively. The levels of serum follicle stimulating and luteinizing hormones were high. An inguinal mass (diameter 9.5 x 7.6) cm was detected on computed tomography. The histopathological diagnosis of this was obtained from laporascopy was composed of ovotestis and there was marked blastic infiltration in this ovotestis which had myeloid markers on flow cytometry. In the immunohistochemical analyses of ovotestis and bone marrow, blasts were positive for LCA, CD-13, CD-33 and CD 68. The cytogenetic analysis of the bone marrow shaved 46 XY karyotype. No response was achieved with combination chemotherapy and the patient died from progressive leukemia. Here we report a rare patient with myelodysplastic syndrome, EMD and hermaphroditism. To our knowledge this is the first case of MDS, EMD and hermaphroditism. PMID- 15360015 TI - Gemcitabine and oxaliplatinum: an effective regimen in a patient with progressive refractory mantle cell lymphoma. AB - We report here a 68-year-old man with progressive refractory Mantle cell Lymphoma stage IV-A, treated with different chemotherapeutic drugs. With no therapeutic options we decided to treat him with oxaliplatinum-gemcitabine regimen, based on the proven efficacy and synergy of these drugs in other tumors and lymphomas. Treatment was well tolerated and patient achieved a maintained complete remission with 6 months of follow up. We consider that this may be a salvage therapeutic option for patients with mantle cell lymphoma. PMID- 15360016 TI - Lethal pulmonary hemorrhage caused by a fulminant Stenotrophomonas maltophilia respiratory infection in an acute myeloid leukemia patient. AB - Stenotrophomonas maltophilia (Sm) pneumonia in immunocompromized hosts is an increasingly common nosocomial infection. Even though resistant to multiple antimicrobials, this gram-negative bacteria usually does not present with a fulminant course leading to a fatal hemorrhagic respiratory infection in neutropenic patients. We report here the case of a 63-year-old woman treated by intensive chemotherapy for acute myeloid leukemia (AML) who presented while severely neutropenic and thrombocytopenic a Sm pulmonary infection with hemoptysis leading to death in 48 h. The bronchoalveolar lavage (BAL) performed shortly before death was highly hemorrhagic and contained a striking amount of extra- and intra-cellular pathogens. Blood and BAL cultures grew S. maltophilia. Post-mortem examination revealed bilateral extensive intra-alveolar hemorrhage (IAH) associated with a great amount of microorganisms and severe bone marrow aplasia was observed without evidence of leukemia residual disease. Sm pneumonia usually does not evolve into such a devastating clinical picture although infections due to the bacteria are known to be associated with high morbidity and mortality. So far, the present observation is the fourth similar case reported in the literature. Even though an early diagnosis and an adequate antibiotic prescription may improve Sm infection prognosis, S. maltophilia proves difficult to eradicate due to a high resistance rate in part intrinsic but also in part acquired. PMID- 15360017 TI - Rituximab-induced remission of a gastric MALT lymphoma. AB - Gastric MALT lymphoma is usually associated with H. pylori infection, and responds to treatment with antibiotics and a proton pump inhibitor. We report a case of H. pylori negative gastric MALT lymphoma. The patient was followed conservatively for 2 years until she developed gastrointestinal bleeding with significant anemia. She was treated with rituximab 375 mg/m2 weekly for four doses, which resulted in a biopsy proven complete remission. Rituximab therapy is a reasonable, well tolerated treatment alternative for MALT lymphomas not associated with H. pylori. PMID- 15360018 TI - Teaching cases from the Royal Marsden and St Mary's Hospitals case 25. A young boy with massive bilateral renal enlargement. PMID- 15360019 TI - Long-term maintenance therapy with weekly chlorambucil in patients with symptomatic or progressive follicular lymphoma. PMID- 15360020 TI - Successful outcome of pregnancy in chronic myeloid leukaemia treated with imatinib. PMID- 15360021 TI - Neutrophil-rich anaplastic large cell lymphoma (NR-ALCL) mimicking lymphadenitis: a study by fine-needle aspiration biopsy. PMID- 15360022 TI - Patient turning schedules: why and how often? PMID- 15360023 TI - Surgical dressings: base purchases on outcomes, not on payment. PMID- 15360024 TI - Uncovering the cause of finger lesions. PMID- 15360025 TI - Diet, diabetes, and wound management: how important is glycemic control? PMID- 15360026 TI - Current concepts in lymphedema management. AB - PURPOSE: To provide physicians and nurses with an overview of the pathophysiology, assessment and diagnosis, and treatment of lymphedema. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in managing patients with lymphedema. OBJECTIVES: After reading the article and taking the test, the participant will be able to: 1. Describe the pathophysiology of lymphedema and the difference between transient and chronic lymphedema. 2. Describe the assessment and diagnosis of lymphedema. 3. Identify treatment options and teaching considerations for patients with lymphedema. PMID- 15360027 TI - Options for off-loading the diabetic foot. PMID- 15360028 TI - Development, validity, reliability, and responsiveness of a new leg ulcer measurement tool. AB - OBJECTIVE: To develop and validate an assessment tool--the Leg Ulcer Measurement Tool (LUMT)--that would be able to detect changes in the appearance of lower extremity ulcers. SUBJECTS: Twenty-two subjects with chronic leg ulcers of various etiologies (arterial, venous, diabetes) participated in the validation study. DESIGN: An interdisciplinary panel consisting of 9 local wound care specialists confirmed content validity. Concurrent criterion validity was determined by correlation of the size domain (1 of 14 clinician-rated domains in the LUMT) with acetate tracing measurement of wound surface area. Reliability was determined using repeated assessments by 4 wound care specialist and 2 inexperienced evaluators; responsiveness was determined using monthly reassessments by a single rater for 4 months. RESULTS: Concurrent criterion validity was r = 0.82. Excellent values of intrarater and interrater reliability (ICC > 0.75) were obtained for total LUMT scores and for many of the 14 individual domains; however, several domains were found to be less reproducible. The LUMT detected change in wound status over time (responsiveness coefficient = 0.84). CONCLUSION: The LUMT can be used by 1 or more assessors, with relatively little previous training, to make reproducible evaluations of lower extremity ulcer appearance and to document change in appearance over time. The LUMT represents a novel assessment tool specifically designed and validated for clinical or research use on chronic leg ulcers. PMID- 15360029 TI - Effects of support surface relief pressures on heel skin blood flow in persons with and without diabetes mellitus. AB - OBJECTIVE: To investigate the effects of pressure relief magnitude on heel blood hyperemia in persons with and without diabetes mellitus. DESIGN: Study participants (1 group of persons with diabetes and 1 group without diabetes) lay on a support surface for 70 minutes with 1 heel on an end cell of a support surface. Cell pressure was computer controlled to be 20 mm Hg during support and 5 or 0 mm Hg during relief. Heel skin blood perfusion was monitored by laser Doppler on the heel and foot dorsum. Heel hyperemia was determined as ratios of skin blood perfusion areas during hyperemia to preloading (AR) and peak hyperemia to mean skin blood perfusion during preload (QR). SUBJECTS: 13 persons with diabetes mellitus (6 females, 7 males; age 65.2 +/- 3.0 years) and no known diabetes-related complications, and 15 persons without diabetes mellitus (7 females, 8 males; age 54.7 +/- 3.1 years) SETTING: University research center. RESULTS: For the nondiabetes mellitus group, hyperemia was significantly greater with complete pressure-relief compared with partial relief (P < .001). In contrast, the diabetes mellitus group showed no significantly increased response to full relief, and the hyperemia achieved during full relief, measured by AR and QR, was significantly less than with the nondiabetes mellitus group. CONCLUSIONS: These results suggest that a diabetes-related reduced microvascular vasodilatory capacity is not exceeded during partial pressure relief, but is exceeded during complete pressure relief. Accordingly, differences in hyperemic response would become unmasked only when maximum hyperemia could be established during complete heel off-loading. This would suggest that a diminished hyperemia during complete off-loading, as found in the present diabetes mellitus group, may be problematic if widely present in the diabetic (or possibly older) population, under conditions in which heel loading occurs for sustained intervals. PMID- 15360031 TI - One mad cow sets off a stampede. PMID- 15360030 TI - Stereologic study of the effects of prostaglandin E2 on the induction of angiogenesis in full-thickness skin autografts. AB - OBJECTIVE: To determine whether prostaglandin E2 improves angiogenesis in full thickness skin grafts. DESIGN: Randomized study SUBJECTS: 20 male rabbits, divided into 2 experimental and 2 control groups. METHODOLOGY: Prostaglandin E2 (experimental groups) or prostaglandin E2 vehicle (control groups) was injected locally into experimental skin autografts once a day for up to 5 days. Five and 10 days after the surgery, the grafts were harvested and after processing, fractional and absolute volumes of the vessels were estimated in the grafted skin using stereologic methods. MAIN RESULTS: Gross appearance of the control and experimental grafts were the same. Qualitative histologic examination of successful grafts in experimental and control groups showed areas of viable epidermis with a negligible inflammatory infiltrate and moderate fibrosis. Blood cells were frequently seen in the vessels under investigation. Histologic slides showed significantly higher mean fractional volume (percent) and absolute volume of the vessels (mm3) per unit volume (mm3) of the grafted skin in the experimental groups than in the control groups. CONCLUSION: Fractional and absolute volume of the vessels were greater in prostaglandin E2-treated grafts than in the control grafts. Prostaglandin E2 appears to increase the volume of vessels in full-thickness skin grafts and can be explored as an agent to improve angiogenesis of the grafts. PMID- 15360032 TI - The impact of a health campaign on health social capital. AB - Referring to literature in sociology, mass communication, and public health, we conceptualize and operationally define "health social capital" and "individual health social capital" and then posit and test a model for its development in response to a public health media campaign. The campaign evaluated here was designed to stimulate behaviors that would provide a more supportive social environment for children and youth, an environment which we consider to be richer in aggregate health social capital. The association model of advertising was employed to explain the development of individual health social capital measures of awareness, attitude, and behavior. With cross-sectional data (1998, n = 614; 1999, n = 1087; 2000, n = 1388), we examine the results for changes in awareness, attitude, and behavior over time and the significant links between these dependent variables and media campaign exposure. The results show significant increases in awareness and attitude, but not in behavior. Structural equation modeling revealed different patterns of influence for newspaper and TV campaign exposure. PMID- 15360033 TI - Rewriting public health information in plain language. AB - Public health materials are often designed to inform and rally the public to spur action and maintain vigilance on important issues to family, work, community, and public policy. Limited access to public health information certainly curtails knowledge and awareness but may also hamper action and civic involvement. A growth in published assessments of health materials indicates an increased interest in the mismatch between the reading level of most health materials and the reading ability of the average adult. However, while several guidebooks offer suggestions for developing new materials, little attention has been given to the process of rewriting materials and grappling with bureaucratic language. We describe, in this case study, a process we used to assess and then rewrite a federally mandated report to consumers about the quality of their water. PMID- 15360034 TI - Knowledge, awareness, and use of the UV index amongst the West Australian public. AB - The ultraviolet (UV) index provides an opportunity to warn people about the degree of hazard that exists on a particular day and subsequently lead to the adoption of sun protective behaviors. While the UV index has been promoted in Australia for some time, little research has been conducted into the understanding and use of the index. As such, a random telephone survey of 501 adults was conducted in December 1999 to gather data into the public's understanding and use of the UV index in Perth, Australia. A high awareness of the UV index existed among the respondents, with 90% indicating they had heard of the term UV index. However, only 5% indicated they had noticed the UV index/forecast for that particular day. The results indicate that a campaign to increase awareness of the UV index by the public is not warranted, but that efforts should be focused on developing tools to communicate behavior change messages linked to the UV index levels. The research was conducted with funding from Healthway, Health Promotion Foundation of Western Australia. PMID- 15360035 TI - Learning from truth: youth participation in field marketing techniques to counter tobacco advertising. AB - In 2000, the American Legacy Foundation (Legacy) launched truth, a national, multi-medium tobacco control social marketing campaign targeting youth age 12-17. This paper provides a brief description of one aspect of that campaign, the truth tour, and compares and contrasts the truth tour with commercial field marketing approaches used by the tobacco industry. The methods used for the tour's process evaluation are also described, and two important lessons learned about using field marketing techniques and using youth to implement field marketing techniques in social marketing campaigns are discussed. Social marketing campaigns that target youth may want to launch field marketing activities. The truth tour experience can inform the development of those efforts. PMID- 15360036 TI - Involving husbands in safe motherhood: effects of the SUAMI SIAGA campaign in Indonesia. AB - Maternal death has implications for the entire family, although few, if any, interventions have addressed the role that the husband could play in his wife's pregnancy and delivery. The Suami SIAGA Campaign in Indonesia was a multi-media entertainment-education intervention, implemented in 1999/2000, that targeted husbands with messages about birth preparedness. The present paper presents the effects of this campaign (1) on the acquisition of new knowledge by husbands about birth preparedness, and (2) on husbands' action toward becoming an alert husband (a Suami SIAGA). When husbands were directly exposed to the messages from the Suami SIAGA campaign, new knowledge gain and birth preparedness activities occurred. However, the interaction of direct exposure to the campaign and the interpersonal communication stimulated by the campaign about Suami SIAGA was an even stronger predictor of knowledge gain and birth preparedness actions. Limitations of the study included post-only measures and a relatively short time period between the intervention and impact evaluation. We suggest that communication campaigns to educate husbands and to stimulate discussion about the innovative concept of birth preparedness may contribute to improved birth outcomes in Indonesia. PMID- 15360037 TI - The impact of emotional tone, message, and broadcast parameters in youth anti smoking advertisements. AB - OBJECTIVES: In the context of controversy regarding the optimal characteristics of anti-smoking advertisements for youth, this study examines the impact on recall and perceived effectiveness of variations in the message, emotional tone, reach and frequency of broadcast, remoteness of broadcast, and characteristics of the adolescent audience such as changes in smoking behavior, ownership of cigarette promotional items, and demographic variables. METHOD: A two-wave longitudinal survey of a population-based sample of 618 Massachusetts youth 12 to 15 years old was carried out in 1993 and 1997. A Generalized Estimating Equation (GEE) approach was used to model the recall and perceived effectiveness of eight advertisements as a function of viewer and ad characteristics. RESULTS: Advertisements featuring messages about serious health consequences which had been independently rated as high in negative emotion were more likely to be recalled and were perceived as more effective by youth survey respondents than ads featuring messages about normative behavior for teens or ads relying on humor. Advertising intensity, while contributing to recall, was negatively related to perceived effectiveness. CONCLUSIONS: This study supports mounting evidence that negative emotion in anti-smoking advertisements is effective with youth audiences. PMID- 15360038 TI - The National Council of State Boards of Nursing. PMID- 15360039 TI - Renaissance in CNS practice. PMID- 15360040 TI - "Mining" data in public databases. PMID- 15360041 TI - Taking the high road: What should you do when an adverse event occurs? Part I. PMID- 15360042 TI - Reduction of perioperative mortality and morbidity with beta blockade: What is known and unknown for the clinical nurse specialist. PMID- 15360043 TI - CNS entrepreneurship: Marketing 101. PMID- 15360044 TI - Genetic practice, education, and research: an overview for advanced practice nurses. AB - PURPOSE/OBJECTIVES: The purpose of this article is to describe how the new genomic era will affect advanced practice registered nurses (APRNs) patient care, education, and research. BACKGROUND/RATIONALE: Given the exponential growth of genetic information and that 9 of the top 10 leading causes of mortality have genetic components (www.cdc.gov), it is imperative to educate advanced practice nurses about this salient topic. DESCRIPTION OF THE PROCESS: Because few APRNs in practice or academia have had formal education on genetics, the first step of nursings' own gene discovery is recognizing that there is an ongoing need to understand state of the science genetic information to gain clinical and educational utility. OUTCOMES: By recognizing APRNs need to know genetics, APRNs will clamor within their workplace for continuing education about this dynamic information. It is critical knowledge for APRNs to classify risk based on family history, target individualized patient prevention and education, modify pharmacologic interventions, and refer when genetic testing is necessary. INTERPRETATION/CONCLUSION: This article stresses the timely relevance of applying genetics and genomics to practice, teaching, and research. IMPLICATIONS FOR NURSING PRACTICE: APRNs need to maintain a place at the genetic table with all healthcare providers by developing strategies to expand this nursing knowledge to their practice, teaching, and research. Nurses need to be cognizant of the keen genetic value of family histories, how risk classification will individualize prevention recommendations, and the exciting role of pharmacogenetics, given many APRNs' prescriptive authority. Our core professional belief that each human is highly unique has probably never been more accurate than with the future in genetic and genomic nursing. PMID- 15360045 TI - Clinical nurse specialists: state of the profession and challenges ahead. AB - Clinical nurse specialists (CNSs) are enjoying a rebirth. As health science and healthcare increase in complexity, it is critical that nurse experts provide leadership to improve patient care, advance nursing practice, and strengthen healthcare delivery systems. CNSs with specialty expertise combined with competencies in patient care, nursing practice, and healthcare delivery systems are essential to translating the products of our vast research enterprise into policies and practices that serve the patient. Many challenges loom. New diseases have emerged; treatments change constantly. We are newly aware of the gravity of the perils posed by natural and man-made disasters. Patients use complimentary and alternative health practices but we do not understand the power and risks of these approaches. Information technology makes possible new approaches to educating nurses and patients and tracking clinical outcomes. CNSs are vital to building a healthcare system that is evidence based, patient centered, outcome effective, safe, ethical, interdisciplinary, and cost-effective. PMID- 15360046 TI - Primary malignant melanoma of the vulva--an aggressive tumor for modeling the genesis of non-UV light-associated melanomas. AB - Malignant melanomas appear in such sun-shielded areas as the vulva, challenging conventional knowledge that they are associated with UV radiation. Based on 1442 patients with vulvar melanomas the tumors' epidemiology, clinical manifestations, histopathology, molecular genetics, treatment strategies, and prognosis were surveyed. Despite their sun-shielded location and rare incidence, vulvar melanomas were, on average, more dense than melanomas on the body surface and nearly the density of melanomas in chronically sun-exposed skin of the head and neck. Vulvar melanomas differed markedly from cutaneous melanomas, as evidenced by histopathological lesions and molecular genetics. Most melanomas were located on the glabrous skin as opposed to the hairy skin within the vulva and differed significantly in biological properties. The prognosis for the patients was poor, and in the 11 largest studies of surgical strategies, none offered a significant survival advantage. Tumor thickness and ulceration were usually significant predictors of (poor) prognosis in multivariate analyses along with macroscopic amelanosis, angioinvasion or DNA non-diploidy in some reports. Clear-cut biological differences between vulvar and cutaneous melanomas and between melanomas within different vulvar sites provide new paths for extensive research on melanomagenesis and for potential therapies. Additionally, studies of vulvar and other extracutaneous melanomas should characterize subgroups of cutaneous melanomas and identify their cause(s), which are apparently not linked to UV radiation. PMID- 15360047 TI - Biodistribution data from 100 patients i.v. injected with 111In-DTPA-D-Phe1 octreotide. AB - The aim of this study was to obtain accurate data on the biodistribution of 111In DTPA-D-Phe1-octreotide in tumour and normal tissues to facilitate dosimetric evaluations. Patients with carcinoid tumours, medullary thyroid carcinoma (MTC), differentiated thyroid tumours, endocrine pancreatic tumour (EPT), breast carcinoma, and various other tumour types were i.v. injected with 111In-DTPA-D Phe-1-octreotide. Tumour and normal tissue samples were collected during surgery 1-35 days later, and the 111In activity concentration determined. Results showed large inter- and intra-individual variations. The 111In concentration was in general higher in carcinoids and some EPT (range 0.33-77% IA/kg) than in MTC and other tumours (0.017-7.8% IA/kg). Tumour-to-blood ratios (T/B) higher than 100 were found in most patients with carcinoids, EPT, renal carcinoma, and neuroendocrine carcinoma (max value 1500), while T/B was below 80 in most other tumours. Normal-tissue-to-blood ratios were in general < or = 10 but higher values were found in liver, kidneys, and spleen. The results presented are important for dosimetric calculations, when radiolabelled octreotide is used for diagnostic or therapeutic purposes. PMID- 15360048 TI - Differential effects of toremifene on doxorubicin, vinblastine and Tc-99m sestamibi in P-glycoprotein-expressing breast and head and neck cancer cell lines. AB - The effect of toremifene on P-glycoprotein-mediated multidrug resistance (MDR) in breast and head and neck cancer cell lines was measured in vitro and in vivo. Pgp expression was low and high, respectively, in drug-sensitive (MCF7-S, KB) and drug-resistant (MCF7-R, MCF7-R1, KBV1) cell lines. Toremifene (7.5 microM) significantly enhanced cytoplasmic and nuclear accumulation of doxorubicin in drug-resistant cells. Toremifene (10 microM) increased the in vitro cytotoxicity of doxorubicin in drug-resistant breast cancer cells (13-fold and 21-fold for MCF7-R and MCF7-R1, respectively) without affecting the sensitivity of MCF7-S cells. Similarly, toremifene (10 microM) caused a 12-fold increase in the sensitivity of KBV1 cells to vinblastine. In contrast, toremifene (5 microM) reduced the net uptake of the radiolabelled Pgp substrate, Tc-99m-sestamibi, in the Pgp-overexpressing cell lines by factors of 0.32 and 0.42 for MCF7-R1 and KBV1 cells, respectively (p < 0.01), and, to a lesser extent, by corresponding factors of 0.89 and 0.86 in the drug-sensitive cell lines (p < 0.05 and p > 0.05, respectively). In nude mice bearing both KB and KBV1 xenograft tumours, significantly higher tumour levels of Tc-99m-sestamibi were recorded in KB tumours compared with KBV1 tumours. After 3 days of treatment with intraperitoneal toremifene (25 mg/kg), tumour levels of Tc-99m-sestamibi were reduced in KB and KBV1 tumours but only statistically significantly for KB tumours. Toremifene is a potent MDR modulating agent with respect to chemotherapeutic agents but has the opposite effect with respect to Tc-99m sestamibi. This finding is of importance in view of the widespread use of Tc-99m sestamibi as an imaging surrogate for a chemotherapeutic agent. PMID- 15360049 TI - ErbB4 is downregulated in renal cell carcinoma--a quantitative RT-PCR and immunohistochemical analysis of the epidermal growth factor receptor family. AB - In the present study the authors evaluated the expression of the EGFR family members ErbB2-4 in renal cell carcinoma (RCC). Thirty-one RCCs were examined for gene expression of ErbB2-4 mRNA by quantitative real-time RT-PCR. For eight of the patients samples of nonneoplastic kidney cortex were also evaluated. Expression of ErbB4 mRNA was analysed in the eight matched tumour and kidney cortex samples by isoform-specific real-time RT-PCR analysis. ErbB4 protein expression was analysed by immunohistochemistry. In summary the results showed that ErbB2 mRNA was downregulated in conventional (clear cell) RCC; ErbB3 mRNA levels were low and heterogeneous in both tumours and kidney cortex; ErbB4 mRNA and protein were strongly downregulated in conventional and papillary RCC. Thus, ErbB2 and ErbB4 are not likely to be oncogenes in the majority of RCCs; instead, the observed downregulations suggest that these receptors might function as tumour suppressors in RCC. PMID- 15360050 TI - Proliferation rate, hormone receptor status and p53 expression in skeletal metastasis of breast carcinoma. AB - Immunohistochemistry was used to analyze ER, PgR, MIB, and p53 of primary breast carcinomas and their skeletal metastases in 43 patients operated on for 47 pathological fractures. Femoral lesions predominated (24), followed by metastases to the spine (13). The median survival time from diagnosis of the primary breast carcinoma was 5.5 (0.5-20) years and from pathological fracture 5 (0-76) months. No clinical characteristics of the primary tumor were prognostic for survival after pathological fracture. Thirty of 47 tissue specimens from skeletal metastases were ER positive and 17 negative. Fourteen of 47 tissue specimens from skeletal metastasis were positive for PgR. Forty-five of the 47 metastases could be evaluated for p53 positivity and 43 tumors showed nuclear staining of varying intensity. In a majority of the cases there was a good correlation between ER, PgR and p53 in the primary tumors and their corresponding metastases. p53 negative primary tumors were associated with longer survival from diagnosis, but also found after pathological fracture. No correlation was observed between ER index or MIB-1 in the skeletal metastases and postoperative survival. PgR positivity in skeletal metastases was associated with longer survival after pathological fracture. This study shows that biological markers such as PgR and p53 provide prognostic information after pathological fracture. The findings should be regarded with caution as there are many confounding factors such as prior chemo- and radiotherapy. Nevertheless, the approach, to gather prognostic data at the time of pathological fracture, warrants further study as expected survival time is paramount in the choice of surgical treatment. PMID- 15360051 TI - Do the results of the process indicators in the Norwegian Breast Cancer Screening Program predict future mortality reduction from breast cancer? AB - Continuous emphases of quality control are required to achieve reduction in mortality from breast cancer as a consequence of breast cancer screening. Results of the process indicators in the first 6 years in 4 counties in the Norwegian Breast Cancer Screening Program are evaluated and will be presented. Data from women who had their initial (n = 173402) and subsequent (n = 220 058) screening provide the basis for the analysis. The breast cancer detection ratio was 3.2 the expected incidence (based on the incidence before the screening started, 1991 1995) among the initially screened women, decreasing to 2.3 among the subsequently screened. The ratio of interval cancer among the initially screened was 0.25 and 0.72 of the expected incidence, 0-12 and 13-23 months after screening, respectively. For those subsequently screened the proportions were 0.22 and 0.64, respectively. More than 50% of the invasive tumors were less than 15 mm in size, and more than 75% were lymph node negative, among both the initially and subsequently screened. The process indicators achieved in the NBCSP are promising as regards future mortality reduction. The incidence of interval cancer 13-24 months after screening is higher than recommended in the European guidelines. PMID- 15360052 TI - Swedish mothers and fathers of a child diagnosed with cancer--a look at their quality of life. AB - Quality of life was investigated among Swedish mothers (n = 118) and fathers (n = 83) of children on (n = 57) and off (n = 68) cancer treatment. Parents completed the Goteborg Quality of Life Instrument, measuring (a) burden of 30 symptoms organized as follows: depression, tension, head, heart-lung, metabolic, musculo skeletal, and gastrointestinal-urinary symptoms, and (b) experience of well-being with regard to 18 items organized as follows: physical, social, and mental well being. Fathers reported higher mental well-being than mothers and more mothers than fathers reported symptoms of depression. Within the same family (n = 70), mothers reported lower mental well-being and more symptoms than fathers. Parents of children on treatment (n = 92) reported lower social and mental well-being and more symptoms of depression than parents with children off treatment (n = 101). The findings suggest that fathers experience a better quality of life than mothers and that parents of children off treatment enjoy a better quality of life than parents of children on treatment. PMID- 15360053 TI - A phase II trial of adjuvant low-dose total body irradiation in non-Hodgkin's lymphoma patients following standard CHOP. AB - Because survival results achieved in aggressive NHL with the standard CHOP are not very satisfactory, we investigated adding adjuvant low-dose total body irradiation (LTBI) to standard CHOP in a phase II trial. Thirty-six patients were included between September 1999 and September 2001. All patients were in documented complete remission (CR) after the end of their standard CHOP. LTBI started 4-6 weeks following the last CHOP course and was given in two courses, each with 4 daily fractions of 0.2 Gy, separated by 2 weeks of rest. Patients with bulky disease received involved-field radiotherapy on initial bulky sites starting 4-6 weeks after the last LTBI fraction. Primary end points were disease free survival (DFS) and overall survival (OS) and the secondary end point was toxicity. The toxicities of LTBI were temporary thrombocytopenia and leucopenia (requiring no transfusions or treatment with growth factors). The 3-year DFS was 61%+/-9% and the overall survival was 87+/-6%. Univariate analysis showed time to achieve CR, and whether the patient got LTBI-induced haematological toxicity to be 2 significant prognostic factors affecting DFS. The use of adjuvant LTBI in patients with aggressive NHL in CR after standard chemotherapy is a feasible, non toxic treatment that is worthy of testing in a future phase III trial. PMID- 15360054 TI - Nodal control and surgical salvage after primary radiotherapy in 1782 patients with laryngeal and pharyngeal carcinoma. AB - The purpose of this study was to evaluate the ultimate neck control after primary radiotherapy and surgical salvage in laryngeal and pharyngeal cancer patients. Some 1782 consecutive patients with squamous cell carcinoma were treated by radiotherapy. At presentation 26% of the patients had metastatic lymph nodes. A total of 298 primary or secondary nodal recurrences were seen, 159 were treated, and 53 (53/298 approximately 18%) were controlled. Isolated N-recurrence was fatal in 2.7% (36/1324) of the N0 patients. Univariate actuarial analysis of nodal control demonstrated that the region of origin, T-classification, T-size, N classification, tumor stage, differentiation, hemoglobin, and radiation time were significant prognostic factors. In a Cox analysis the independent significant parameters were gender, region of origin, N-classification, and differentiation. The conclusions were that in patients with nodal recurrence a little over half were treated and of these a third of the nodal recurrences were controlled. Significant prognostic factors in multivariate analysis were gender, region of origin, N-classification, and tumor differentiation. PMID- 15360056 TI - Effect of heterogeneity in radiosensitivity on LQ based isoeffect formalism for low alpha/beta cancers. AB - Change of fractionation for external beam radiation therapy based on linear quadratic (LQ) formalism assumes that a single alpha/beta is sufficient to characterize tumour response to dose fractionation. In reality, both inter patient and intra-tumour heterogeneity might affect the applicability of isoeffectiveness formalism. The impact of heterogeneity on recently proposed hypofractionation schemes for the prostate has been analysed. The alpha/beta ratio was assumed to be Gaussian distributed with a mean value of 1.5 Gy. Gaussian and lognormal distributions for alpha were modelled. TCP model parameters were adjusted to lead to TCP = 0.80 for 70 Gy at 2 Gy per fraction. TCP loss from heterogeneity and doses required to restore TCP = 0.80 were calculated. The effect of heterogeneity was moderate. Doses to restore TCP = 0.80 in most cases were less than 1 Gy. The largest TCP loss was 4%. The difference between predictions of single alpha/beta and heterogeneity models is too small to be detected in a clinical trial. PMID- 15360055 TI - Transient CEA increase at start of oxaliplatin combination therapy for metastatic colorectal cancer. AB - In general a rising carcinoembryonic antigen (CEA) level means tumor progression. We observed a transient increase in CEA level despite objective response among patients receiving chemotherapy for metastatic colorectal cancer. This surge phenomenon has not previously been described for patients with metastatic colorectal disease. CEA was measured every second week in 27 patients receiving oxaliplatin, 5-fluororuracil, and folinic acid as first-line therapy against metastatic colorectal cancer. Four patients (15%, 95% CI 5-31%) met the criteria for therapy-induced CEA surge. The time of reaching maximum CEA level varied from 13 to 56 days. Median rise in CEA from baseline was 263% (range 24-632%). An initial rise of CEA during chemotherapy in colorectal cancer patients may therefore not always indicate progression of disease but may be a transient CEA surge in patients responding to chemotherapy. PMID- 15360057 TI - Long-term survival of acute leukaemia--one patient's story--ethical and historical considerations. PMID- 15360058 TI - Exhaled nitric oxide in childhood asthma--time to use inflammometry rather than spirometry? PMID- 15360059 TI - Relationship between asthma severity and obesity. AB - The prevalence of both obesity and asthma has risen in recent years. We sought to investigate whether obesity may be related to asthma. We undertook a retrospective medical record review of patient records at an inner-city academic asthma center. Obesity was defined as a body mass index (BMI) greater than 30. Asthma severity was defined by using the National Heart Lung and Blood Institute 1997 guidelines. Adults with a history of cigarette smoking or other lung disease were excluded. A total of 143 individuals aged 18-88 with a mean age of 43.9 met the entry criteria. There were 113 females and 30 males. Seventy-two percent of the sample was obese. The Spearman correlation coefficient showed a linear relationship between asthma severity and BMI (r = 0.40, p < 0.0001). Females with asthma were significantly more overweight than males, mean BMI 35.9 vs. 32.14, respectively (p = 0.01). The prevalence of obesity in the 13 patients on long term oral corticosteroids was 100%. Prevalence of obesity increases with increasing asthma severity in adults. The association of asthma severity with obesity suggests that obesity may be a potentially modifiable risk factor for asthma or asthma-like symptoms. PMID- 15360060 TI - A right-sided aortic arch misdiagnosed as asthma since childhood. AB - Congenital malformation of the aortic arch complex occurs in 3% of humans. These vascular aberrations result from embryonic structures that persist when there is incomplete atrophy or regression during normal development. Typically, anomalies of the aortic arch do not cause respiratory symptoms. However, in some individuals, the unusual position of the remnant vessels may compress the airway and cause airflow limitation. As might be expected, a symptomatic vascular anomaly is most often clinically apparent in childhood. We report an adult male with exertional dyspnea who had been diagnosed with asthma since childhood. After his symptoms proved refractory to newer asthmatic therapy, he was referred for further evaluation. Bronchial hyperactivity was not present with objective testing, and compression of his trachea by a right-sided aortic arch was confirmed with appropriate imaging studies. This report illustrates the need to confirm the diagnosis of asthma with objective measures of airflow and reminds the clinician that a congenital vascular aberrancy may initially elude diagnosis. PMID- 15360061 TI - Knowledge, attitudes, and behavior of the parents of asthmatic children in Maputo. AB - Asthma is increasing in frequency worldwide. The education of affected children and their parents is fundamental for the management of the disease. The aim of our study was to describe the knowledge, attitudes, and behavior of the parents of asthmatic children. We studied 152 adults accompanying asthmatic children to consultations in the pediatrics department of Maputo Central Hospital (Mozambique). In general, knowledge about asthma was poor: 11% of the adults thought that asthma was contagious and transmitted from person to person, and 4% thought that it was transmitted by contaminated food. More than half the parents thought that the child could not lead a normal life even during the periods between attacks. A large proportion of the parents thought that asthma could be cured by medical treatment in 7% of cases and by alternative treatment in 43% of cases. The precipitating factors were well identified by the adults, but they had an inaccurate perception of the symptoms of an asthma attack. The actions of the various classes of drugs used were poorly understood. Education programs for both the children and their parents should be developed to improve the management of asthmatic children. Surveys of this type facilitate the targeting of such programs. PMID- 15360062 TI - The ECP/Eo count ratio in children with asthma. AB - We hypothesized that the serum eosinophil eationic protein (ECP) concentration to peripheral blood eosinophil count ratio (ECP/Eo ratio), reflecting active eosinophils, could better correlate with asthma severity in asthmatic patients, than each of these parameters alone. One hundred twenty children with mild to moderate persistent stable asthma were included into the study. At the first visit, previous asthma medications were withheld and patients were administered beta-2 agonists "as needed." At the second visit peripheral blood eosinophil count, serum ECP, sIL-2R, and sICAM-1 were measured, and spirometry and histamine challenge tests were performed. During the study, patients filled daily diary cards to assess symptoms score. One hundred seventeen patients completed the study. The univariate logistic regression analysis showed that asthma severity is related to PC20H, ECP, ECP/Eo ratio, sIL-2R, and sICAM-1. In general, patients with higher level of ECP, ECP/Eo ratio, sIL-2R, sICAM-1 and with lower PC20H exhibited the higher risk of moderate asthma. Multivariate regression analysis showed that only PC20H and ECP/Eo ratio were the best predictors of asthma severity; higher PC20H (1 mg/mL change) slightly decrease (OR = 0.656; 95% CI: 0.44-0.99) and higher ECP/Eo ratio (0.1 pg/cell change) increase (OR = 1.84; 95% CI: 1.02-3.34) a risk of moderate asthma. These data show that the ECP/Eo ratio is a better and more useful marker than ECP or peripheral blood eosinophil count separately in assessing asthma in children. PMID- 15360063 TI - Levamisole: might it be used in treatment and prevention of atopic diseases? AB - As an antihelmintic and Th1-biased immunostimulant, levamisole has been used to restore impaired cell mediated immunity. We sought to explore whether the Th1 driving effect of levamisole may also have an influence on the course of allergic diseases, by shifting the Th2 dominant immunity more toward Th1-mediated response. BALB/c mice were sensitized intraperitoneally on days 0, 7, and 15 with ovalbumin (OVA). After the sensitization, they were challenged intranasally with OVA once a day for 6 consecutive days. Levamisole (2.5 mg/kg) was administered orally three times a week during sensitization and challenge. After the last challenge, differential cell counts were performed, and IL-4 and IFNgamma levels were measured in the bronchoalveolar lavage fluids (BALF). Serum total IgE level was determined, and lungs were examined histologically. The present study establishes that mice administered with oral levamisole gave significantly lower IL-4 levels on sensitization with OVA; however, IFNgamma production, eosinophil infiltration, and serum IgE levels remained unaffected. In conclusion, use of levamisole may have important implications in the management of the allergic inflammation. PMID- 15360064 TI - Airway hyperresponsiveness by methacholine challenge testing following negative exercise challenge. AB - BACKGROUND: Exercise challenge testing (ECT) to diagnose exercise-induced bronchospasm has been demonstrated to be an insensitive screening test to demonstrate the presence or absence of airway hyperreactivity. Previous studies have not compared this procedure to methacholine challenge testing (MCT) in a clinical setting. OBJECTIVE: To determine the frequency of positive MCT in subjects with exertional dyspnea, normal baseline spirometry, and negative ECT. METHODS: Observational study of 215 military patients at an Army Community Hospital referred for evaluation of exertional dyspnea with normal baseline spirometry. Subjects were further evaluated with ECT on a graded treadmill with pre- and postexercise spirometry. Those without evidence of bronchial hyperreactivity as defined by a 15% decrease in FEV1 postexercise were evaluated with methacholine challenge testing (MCT). RESULTS: Two hundred ten military subjects were evaluated. Eighty-two patients underwent ECT as the only method of bronchoprovocation testing with 25 positive tests (57 were negative but not referred for further testing). The remaining 128 patients with a negative ECT underwent MCT. Seventy-six (59%) had a negative MCT and 52 (41%) had a positive MCT. Of the positive MCT studies, 74% were positive at a methacholine concentration of 2.5 mg/mL or less. CONCLUSIONS: Our study demonstrates that a significant number of patients being evaluated for exertional dyspnea will have a positive MCT after a negative ECT. Our findings lead us to question the utility of ECT as an initial diagnostic test for the exercise-induced bronchospasm. PMID- 15360065 TI - A cost-effectiveness analysis of a peak flow-based asthma education and self management plan in a high-cost population. AB - BACKGROUND: Asthma education and action plans (AP) have been recognized as important components in the optimal management of asthma. Studies have differed on the importance of a peak flow-based self-management plans in reducing health care costs and use due to asthma exacerbation. OBJECTIVE: To analyze the cost effectiveness of peak flow-based action plans in reducing costs associated with ER visits and hospitalizations due to acute asthma exacerbation in a population of high-risk and high-cost patients, defined as patients with moderate to severe asthma with a history of recent urgent treatment in the ER or hospitalization due to asthma. METHODS: A literature review of randomized clinical trials comparing peak flow-based (PFB) action plans, symptom-based (SB) action plans, and usual care/no action plan (NAP) was performed. Probability values regarding the effectiveness of each alternative (as measured by increase/decrease in ER visits and hospitalizations over a 6-month period) were derived. Incremental cost effectiveness and cost-benefit ratios were calculated for each alternative. Sensitivity analyses were performed. RESULTS: For high-risk and high-cost asthma patients, our analysis revealed that the most cost-effective alternative for reducing ER visits was a peak flow-based self-management plan. The peak flow based self-management program had an incremental cost-effectiveness (C/E) ratio of $ 60.57 per ER visit averted compared to usual care/NAP and a C/E ratio of $31.46 compared to the SB-AP. The PFB-AP was also the most cost-effective in reducing asthma hospitalization costs with an incremental C/E ratio of $300 per hospitalization prevented, compared with usual care and a C/E ratio of $311, compared to a SB-AP. Analysis yielded a cost-benefit ratio of 13.79 for the PFB AP compared to NAP; the SB-AP had a cost-benefit ratio of 11.53 compared to NAP. CONCLUSION: Cost-effectiveness and cost-benefit analyses reveal that for high cost patients, a peak flow-based asthma education and self-management plan program is the most cost-effective alternative in reducing costs associated with ER visits and hospitalizations due to asthma exacerbation. Further refinements to this cost-effectiveness analysis including measuring changes in drug use and costs and patients' productivity losses need to be pursued and may demonstrate additional cost-savings due to peak flow-based asthma education plans. PMID- 15360066 TI - Airways hyperresponsiveness in high school athletes. AB - Adult athletes have a higher prevalence (11%-50%) of exercise-induced bronchoconstriction (EIB) and airways hyperresponsiveness (AHR) than the population at large (7%-11%): reports describing EIB/AHR in adolescent athletes are scant. HYPOTHESES: 1) a minimum AHR prevalence of 20% would be revealed in a group of high school athletes; 2) demographic data would predict AHR; 3) AHR positive athletes would preferentially choose low ventilation sports. Eucapnic voluntary hyperpnea (EVH) was used to test for AHR in 23% of all athletes (79 of 343) of a midwestern high school. The AHR was defined by at least a 10%, 20%, or 25% decline in FEV1, FEF25-75, or PEFR at 1, 5, 10, or 15-min post-EVH, respectively. RESULTS: 30 of 79 (38%) tested positive for AHR; demographic data tended to predict AHR, as correlations between the total number of years exercised with the greatest decline in FEV1 and the total number of days exercised with the greatest decline in FEV1 following the EVH challenge tended to be significant (r = 0.354; p = 0.055 and r=0.314; p = 0.091, respectively); and 69% of AHR-positive students played only low ventilation sports. CONCLUSION: AHR prevalence was 38% in athletes of a midwestern high school; demographic data tended to predict AHR; those with AHR preferentially play low ventilation sports. PMID- 15360067 TI - Efficacy and safety of inhaled corticosteroids in combination with a long-acting beta2-agonist in asthmatic children under age 5. AB - The incidence of asthma in children under age 5 is higher than in any other segment of the population. Current NAEPP guidelines recommend treatment of some asthmatics in this age group with the combination of an inhaled corticosteroid and a long-acting beta2-agonist even though this practice has never been studied with children younger than 4. This retrospective study analyzes the efficacy and safety of a combination of fluticasone propionate (FP) and salmeterol (SA) in children under 5. Fifty patients who started using FP/SA before the age of 60 months were included in the analysis. To determine efficacy, we tracked the change in emergency room visits, hospitalizations, and the frequency of wheezing as a result of treatment. Emergency room visits were reduced from 78 to 5 (p<0.001), hospitalizations were reduced from 43 to 2 (p<0.001) and frequency of wheezing, daily, weekly, or monthly, was also reduced significantly (p<0.003). In terms of safety, there was only a 3.4% reduction in height percentile (p=0.37). Combination therapy is highly efficacious and safe for asthmatics under the age of 5. A well-designed prospective study is necessary to further evaluate the benefits and risks of this treatment method. PMID- 15360068 TI - An evaluation of the asthma intervention of the New York State Healthy Neighborhoods Program. AB - BACKGROUND: The Healthy Neighborhoods Programs (HNP) are funded by the Federal Preventive Health and Health Services block grants and administered by the New State Department of Health (NYSDOH). Eight county and local health departments are funded for 3 years for a total of $1.25 million per year to target households at risk for environmental health and safety hazards. The HNP asthma intervention uses home visits to identify asthmatics, assess asthma morbidity and management, and identify environmental asthma triggers. Outreach workers provide education about asthma, referrals, and controls for asthma triggers. The purpose of this evaluation was to assess the impact of the HNP asthma intervention for the 1997 1999 funding cycle and for the first year of the 2000-2002 funding cycle. Because of changes in reporting requirements across the funding cycles, the findings for 1997-1999 and 2000 were analyzed separately. METHODS: We analyzed one final outcome measure, the rate of self-reported hospitalizations (admissions and emergency room visits), and three intermediate outcome measures (the percent of homes with cockroaches, the percent of asthmatics with a written management plan, and the percent of asthmatics using a peak flow meter). We also estimated the net savings resulting from a reduction in hospital admissions due to asthma. RESULTS: For the 1997-1999 funding cycle, the average hospitalization rate was 86 hospitalizations per 100 asthmatics per year at the intervention visit (i.e., the baseline rate) and 44.5 at the 1-year follow-up, a decrease of 48%. This was a larger decrease than the 24% average annual decrease in the baseline rates during the 3 years. In 2000, there were 96 hospitalizations per 100 asthmatics per year at the intervention visit and 25 at the one-year follow-up, a decrease of 74%. In 2000 there were about 110 fewer hospital admissions thought to be due to the net effects of the HNP intervention, resulting in an estimated gross savings of $905,300 (110 x $8,230). After subtracting the estimated cost of the asthma intervention ($624,683), the net savings were $280,617. CONCLUSION: New York's HNP seems to be succeeding in reducing asthma morbidity as measured by the hospitalization rates among asthmatics who have received the intervention. The HNP was successful in decreasing cockroach infestation in HNP homes. A standardized evaluation methodology and instrument are necessary to conduct a more rigorous evaluation of the HNP. PMID- 15360069 TI - The shocking truth about verbal abuse. PMID- 15360070 TI - Bowel care is fundamental to nursing. PMID- 15360071 TI - Is holistic care always the best approach? PMID- 15360073 TI - Silence the abuse. PMID- 15360074 TI - How nurse intervention is tackling child obesity. PMID- 15360075 TI - 'I call it the rock and roll of nursing'. PMID- 15360076 TI - The rising incidence of antibiotic-resistant ESBL-producing E. coli. AB - Extended-spectrum beta-lactamase (ESBL) Escherichia coil produces enzymes that enable it to develop resistance to antibiotics. Its prevalence in the UK is increasing, and the Health Protection Agency is investigating whether it has contributed to a number of deaths. While many people may be asymptomatic carriers of ESBL-producing E. coli it can be extremely harmful to the very young, the very old and those with compromised health and other risk factors. Excellent infection control is necessary to prevent its spread in health care facilities, while restrictions on the use of antibiotics may be necessary to prevent its increasing resistance. PMID- 15360077 TI - What you need to know about... thrombosis. PMID- 15360078 TI - Supporting the management of diabetes during periods of illness. PMID- 15360079 TI - The use of pastoral and spiritual support in bereavement care. AB - For many people, the death of a loved one can result in feelings of shock, numbness or denial, even though they may have been expecting it for some time. Pastoral and spiritual care is a vital part of an authentic, holistic health care service. Nurses need to be aware of the needs of relatives and friends of the deceased and understand their role in practical matters such as death certificates, cremation forms and last offices. PMID- 15360081 TI - The effective management and administration of premedication. AB - Nurses play an essential role in preparing patients for surgical procedures, whether the operation is planned or an emergency. Part of this preparation may include administration of premedication. Government initiatives such as The NHS Plan (Department of Health, 2000) and Essence of Care (DoH, 2001) emphasise the need to get the fundamental aspects of nursing care right, and nurses have a professional responsibility to deliver evidence-based care (NMC, 2002). Additionally, the publication of Building a Safer NHS for Patients: Improving Medication Safety (DoH, 2004) aims to improve patient safety by reducing the incidence of drug errors. This article looks at why pharmacological intervention is no longer a routine preoperative preparation. It will discuss drugs that are used to aid anaesthesia and those used as prophylaxis to aid postoperative recovery, consider the ways in which premedication can now include psychological interventions, and look at the nurse's role. PMID- 15360080 TI - Using psychosocial interventions within a high-security hospital. AB - Psychosocial interventions have been used for many years in caring for people with mental health problems. The few interventions that have a recognised evidence base are cognitive behavioural therapy (CBT), family interventions, and education programmes for patients and carers. The process of implementing these interventions in a forensic setting will be described and recommendations will be made for future practice. PMID- 15360082 TI - Challenges in the field of HIV nursing. Interview by Rachel Leslie. PMID- 15360083 TI - Pushing the boundaries of nursing. PMID- 15360084 TI - Bystander effect and adaptive response in C3H 10T(1/2) cells. AB - PURPOSE: To address the relationship between the bystander effect and the adaptive response that can compete to impact on the dose-response curve at low doses. MATERIALS AND METHODS: A novel radiation apparatus, where targeted and non targeted cells were grown in close proximity, was used to investigate these phenomena in C3H 10T(1/2) cells. It was further examined whether a bystander effect or an adaptive response could be induced by a factor(s) present in the supernatants of cells exposed to a high or low dose of X-rays, respectively. RESULTS: When non-hit cells were co-cultured for 24 h with cells irradiated with 5 Gy alpha-particles, a significant increase in both cell killing and oncogenic transformation frequency was observed. If these cells were treated with 2 cGy X rays 5 h before co-culture with irradiated cells, approximately 95% of the bystander effect was cancelled out. A 2.5-fold decrease in the oncogenic transformation frequency was also observed. When cells were cultured in medium donated from cells exposed to 5 Gy X-rays, a significant bystander effect was observed for clonogenic survival. When cells were cultured for 5 h with supernatant from donor cells exposed to 2 cGy and were then irradiated with 4 Gy X-rays, they failed to show an increase in survival compared with cells directly irradiated with 4 Gy. However, a twofold reduction in the oncogenic transformation frequency was seen. CONCLUSIONS: An adaptive dose of X-rays cancelled out the majority of the bystander effect produced by alpha-particles. For oncogenic transformation, but not cell survival, radioadaption can occur in unirradiated cells via a transmissible factor(s). PMID- 15360085 TI - Differential DNA double strand break fixation dependence on poly(ADP ribosylation) in L5178Y and CHO cells. AB - PURPOSE: To investigate the role of poly(ADP-ribosylation) in DNA double-strand break repair and fixation in murine lymphoma L5178Y (LY) sublines, LY-R and LY-S, and a pair of Chinese hamster ovary lines: wild-type and mutant xrs6 cells, that have differences in repair competence and degree of radiosensitization with poly(ADP-ribosylation) inhibitors. MATERIALS AND METHODS: Cells (asynchronous, logarithmic phase) were pre-incubated with 2 mM aminobenzamide at 37 or 25 degrees C, X-irradiated with 10 Gy and allowed to repair DNA breaks for 15, 60 and 120 min at 37 or 25 degrees C. The remaining double-strand break were estimated by the neutral comet assay. RESULTS: At 37 degrees C, no effect of AB treatment on the repair kinetics was observed either in xrs6 or Chinese hamster ovary (wild-type) cells. In contrast, aminobenzamide decreased the repair of double-strand break in the LY-S line but not the LY-R line, in agreement with the previously observed radiosensitization of LY cells by poly(ADP-ribosylation) inhibition. However, double-strand break rejoining in the repair competent cell lines, Chinese hamster ovary and LY-R, also was affected by aminobenzamide when the post-irradiation incubation was carried out at 25 degrees C. Analysis of these results together with earlier data on LY-S cells have been interpreted in terms of Radford's model of radiation damage fixation. CONCLUSION: The reported results indicate that poly(ADP-ribosylation) can be an important modulator of the conversion of DNA damage to lethal events. PMID- 15360086 TI - Radicicol potentiates heat-induced cell killing in a human oesophageal cancer cell line: the Hsp90 chaperone complex as a new molecular target for enhancement of thermosensitivity. AB - PURPOSE: To examine the ability of a heat shock protein 90 (Hsp90) chaperone complex inhibitor, radicicol, to modify thermal response and heat-induced cell killing, and to clarify the underlining mechanisms. MATERIALS AND METHODS: A human oesophageal cancer cell line (TE-1), with a mutant p53 gene, was used. To examine the effect of radicicol on heat-induced cell killing, radicicol at a concentration of 100 nM was incubated with the cells for 7 h during heat treatment. Changes in the expression of proteins were examined by Western blot and immunofluorescence analysis. RESULTS: Radicicol in combination with heat synergistically potentiated heat-induced cellular killing despite an increase in the expression of Hsp72 and Hsp27 caused by radicicol. Heat alone activated Raf-1 and p42/p44 extracellular signal-regulated kinase (Erk), and heat in combination with radicicol inhibited the activation of Raf-1 and p42/p44 Erk through reduced binding of Raf-1 to Hsp90. Phosphorylation of Akt was also decreased by radicicol. CONCLUSIONS: The Hsp90 chaperone complex inhibitor, radicicol, potentiated heat-induced cellular killing, and inhibition of p42/p44 Erk and Akt activation rather than modification of Hsp expression might be involved in enhancing cellular thermosensitivity. Results suggest that the Hsp90 chaperone complex could be a new molecular target for the modification of the cellular response to heat. PMID- 15360087 TI - Occupational exposure to ionizing radiation has no effect on T- and B-cell total counts or percentages of helper, cytotoxic and activated T-cell subsets in the peripheral circulation of male radiation workers. AB - PURPOSE: To investigate changes in immune cell subsets in the peripheral circulation of a male population occupationally exposed to ionizing radiation. MATERIALS AND METHODS: Peripheral blood samples were taken from 194 male workers with cumulative exposures of >200 mSv (mean exposure 331.5 mSv, mean age 51 years) and from a reference population of 131 male workers with cumulative exposures of <27.5 mSv (mean exposure 13.9 mSv, mean age 47 years). Samples were analysed by flow cytometry for T- and B-cell total counts and for the T-cell subset percentages of CD4+ (helper T-cells), CD8+ (cytotoxic T-cells) and CD3+/HLA-DR+ (activated T-cells). RESULTS: Comparison of the >200 and <27.5 mSv exposure groups using linear regression analysis showed no statistically significant differences between the two groups for T-cell total count, B-cell total count or for percentages of the T-cell subsets CD4+, CD8+ or CD3+/HLA-DR+ and CD4+:CD8+. However, statistically significant increases in both T- and B-cell total counts were observed within the two exposure groups and data pooled from both groups when non-smokers (never and ex-smokers) were compared with current smokers. For pooled data T-cell total count increased in smokers by 35% (p=0.0001) and B-cell total count increased by 37% (p=0.0004). CONCLUSIONS: No significant immunological effects were observed in male radiation workers with cumulative exposures of >200 mSv when compared with a reference population with cumulative exposures of <27.5 mSv, although highly significant increases in both T- and B-cell total counts were observed in smokers compared with non-smokers. PMID- 15360088 TI - Is sensitization with nicotinamide and carbogen dependent on nicotinamide concentration at the time of irradiation? AB - PURPOSE: To determine whether tumour radiosensitization and the therapeutic benefit of administering carbogen with nicotinamide depend upon irradiating at the time of peak drug concentration. MATERIALS AND METHODS: Local tumour control of CaNT tumours in CBA mice and acute skin reactions in albino WHT mice were assessed after treatment with 10 X-ray fractions in air, carbogen alone or combined with 0.1, 0.2 or 0.5 mg g(-1) nicotinamide, injected 15, 30 or 60 min before irradiation. Plasma and tumour drug pharmacokinetics were performed. RESULTS: Nicotinamide was rapidly taken up into tumours; a six- and threefold higher concentration was obtained with 0.5 mg g(-1) compared with 0.1 and 0.2 mg g(-1), respectively. Tumour, but not skin, radiosensitization increased as the dose of nicotinamide increased (p = 0.03), but at each dose level there was no significant difference in radiosensitivity when irradiations were done at or after the time of peak concentration. An almost eightfold increase in plasma levels increased tumour enhancement ratios from 1.74 to 1.92 (p < 0.0001). In tumours all schedules gave significant enhancement relative to carbogen alone (p < or = 0.04). CONCLUSIONS: Tumour and skin radiosensitivity was independent of time of nicotinamide administration. Higher drug concentrations were not mirrored by proportionally higher enhancement ratios. Lower plasma levels than previously suggested significantly enhanced tumour radiosensitivity relative to carbogen alone. The clinical implications of these findings are discussed. PMID- 15360089 TI - Chromosome spatial clustering inferred from radiogenic aberrations. AB - PURPOSE: Analysing chromosome aberrations induced by low linear energy transfer (LET) radiation in order to characterize systematic spatial clustering among the 22 human autosomes in human lymphocytes and to compare their relative participation in interchanges. MATERIALS AND METHODS: A multicolour fluorescence in situ hybridization (mFISH) data set, specifying colour junctions in metaphases of human peripheral blood lymphocytes 72 h after in vitro exposure to low LET radiation, was analysed separately and in combination with previously published results. Monte Carlo computer simulations and mathematical modelling guided data analysis. RESULTS AND CONCLUSIONS: Statistical tests on aberration data confirmed two clusters of chromosomes, [1, 16, 17, 19, 22] and [13, 14, 15, 21, 22], as having their members being on average closer to each other than randomness would predict. The first set has been reported previously to be near the centre of the interphase nucleus and to be formed mainly by gene-rich chromosomes, while the second set comprises the nucleolus chromosomes. The results suggest a possible interplay between chromosome positioning and transcription. A number of other clusters suggested in the literature were not confirmed and considerable randomness of chromosome-chromosome juxtapositions was present. In addition, and consistent with previous results, it was found that chromosome participation in interchanges is approximately proportional to the two-thirds power of the DNA content. PMID- 15360090 TI - Comparison of DNA damage and repair following radiation challenge in buccal cells and lymphocytes using single-cell gel electrophoresis. AB - PURPOSE: To develop a reproducible single-cell gel electrophoresis assay for DNA damage and repair in buccal mucosa and sublingual exfoliated cells. MATERIALS AND METHODS: Buccal mucosa and sublingual cells and lymphocytes from six individuals (three males, three females, aged 34-45 years) were challenged with increasing doses of gamma-rays. DNA strand breaks and DNA repair were measured using the single-cell gel electrophoresis assay. RESULTS: Baseline DNA strand breaks were significantly greater in buccal mucosa and sublingual cells compared with lymphocytes. Buccal mucosa and sublingual cells did not differ from each other with respect to induction of DNA strand breaks by 2 or 4 Gy gamma-rays. However, they showed a smaller increase in gamma-ray-induced DNA strand breaks compared with lymphocytes (32-53% less than lymphocytes; ANOVA p<0.0001). Unlike lymphocytes, which repaired 83% of DNA strand breaks, buccal mucosa and sublingual cells exhibited only a minimal capacity for DNA repair (approximately 0-14% of the level in lymphocytes). CONCLUSIONS: Buccal mucosa and sublingual cells exhibit an apparent resistance to the expression of radiation-induced DNA strand breaks in vitro and an apparent lack of DNA strand break repair in the single-cell gel electrophoresis assay. PMID- 15360091 TI - Time benefit in the assessment of recurrences following fractionated radiotherapy in an experimental tumour system using positron-emission tomography with 18F fluorodeoxyglucose. AB - PURPOSE: To determine the sensitivity and specificity of 18F-fluorodeoxyglucose positron-emission tomography (FDG-PET) in the diagnosis of R1H tumours after fractionated radiotherapy, and the dependency of sensitivity and specificity on time after therapy. In addition, the time benefit of FDG-PET concerning early recognition of recurrences after fractionated radiotherapy was assessed. MATERIAL AND METHODS: Subcutaneously growing rat rhabdomyosarcoma R1H tumours were irradiated by applying total doses of 80 or 85 Gy after reaching a start volume of 0.8 cm3. Twenty animals were treated. Tumour volume was determined twice a week. FDG-PET was performed weekly before, during and for 6 months after therapy using a conventional full-ring whole-body PET scanner. In total, 600 PET results were evaluated qualitatively using a six-scale score. PET results and actual tumour volumes were compared. The sensitivity and specificity of tumour detection by PET was calculated for different times after the onset of therapy. The optimal score for tumour detection and the influence of time after therapy on the quality of PET (time benefit) was evaluated using receiver-operating characteristics. RESULTS: After irradiation, 8/20 tumours (40%) were locally controlled, while 12/20 recurred. In this tumour model, evidence of relapse is assured when a volume of 0.1 cm3 is reached. Sensitivity of tumour diagnosis by PET increases with time, i.e. with the volume of recurrent tumours after the onset of therapy, mounting to > 0.95 after 100 days. Specificities of 0.95-1.0 were determined after therapy, showing no increase with time. Tumour diagnosis by PET is highly accurate when performed 80 days after the start of treatment. On average, tumours were recognized by PET on 31, 62, 74 and 81 days (median) before approaching volumes of 0.2, 0.5, 0.8 or 1.0 cm3, respectively. CONCLUSION: An experimental system was implemented that allows reproducible detection of recurrent R1H tumours after radiotherapy using FDG-PET. The usefulness of PET as a diagnostic test for R1H tumours is very good and a reliable resolution for PET is demonstrated for volumes < 1 cm3. The results indicate that FDG-PET enables early recognition of recurrences after fractionated radiotherapy. PMID- 15360092 TI - Streptococcus pneumoniae: epidemiology and patterns of resistance. AB - Streptococcus pneumoniae is a leading cause of bacterial pneumonia, meningitis, otitis media, and sinusitis; it results in significant morbidity and mortality in patients with pneumonia and meningitis. The pneumococcus is a common colonizing bacterium in the respiratory tract; it is especially common in the respiratory tracts of children, where it is frequently exposed to antimicrobial agents. This exposure can lead to resistance. Penicillin nonsusceptibility is found in nearly 40% of strains causing disease in adults, although often these cases are treatable with appropriate dosing regimens of many oral and parenteral beta lactam agents. In the United States resistance to macrolides is widespread- averaging approximately 28%--but geographically variable, ranging from 23% in the northwest to 30% in the northeast. Resistance to tetracyclines and trimethoprim sulfamethoxazole are reported in approximately 20% and 35% of isolates, respectively, and resistance to multiple classes of agents is increasingly common. Amoxicillin, amoxicillin-clavulanate, respiratory fluoroquinolones, and clindamycin are currently the most effective agents for treatment of respiratory tract infections caused by S pneumoniae, with >90% of isolates in the United States being susceptible. Vancomycin is the only agent against which resistance has not emerged. Patient groups that are at increased risk for developing resistant pneumococcal infections have been identified and include patients with malignancies, human immunodeficiency virus infection, and sickle-cell disease. Judicious use of antimicrobials is the key to preventing the emergence of further resistance, particularly as few new classes of agents are likely to become available for clinical use in the short term. PMID- 15360093 TI - Overview of newer antimicrobial formulations for overcoming pneumococcal resistance. AB - The pharmacokinetic (PK) and pharmacodynamic (PD) profile of an antimicrobial agent provides important information that can be used to maximize bacteriologic and clinical efficacy, minimize selective pressure for the development of antimicrobial resistance, and determine an optimal dosing regimen. Judicious selection of an antimicrobial based on local susceptibility data and PK and PD parameters is imperative in this era of increasing resistance among Streptococcus pneumoniae, a leading cause of community-acquired respiratory tract infections. The beta-lactam antimicrobials display time-dependent bacterial killing with minimal to no persistent effects. Ketolides and fluoroquinolones display concentration-dependent bacterial killing, and tetracyclines and macrolides display time-dependent killing. All have prolonged persistent effects (e.g., postantibiotic effect) that retard or prevent bacterial regrowth when free drug levels fall below the minimum inhibitory concentration (MIC). New high-dose and/or extended-release formulations of traditional antimicrobials have been added to the current armamentarium for treatment of community-acquired respiratory tract infections. These formulations include amoxicillin-clavulanate potassium powder for oral suspension 90/6.4 mg/kg per day divided every 12 hours (Augmentin ES-600; GlaxoSmithKline, Research Triangle Park, NC), amoxicillin clavulanate potassium extended-release tablets 2 x 1,000 mg/62.5 mg every 12 hours (Augmentin XR; GlaxoSmithKline), clarithromycin extended-release tablets 2 x 500 mg once daily (Biaxin XL; Abbott Laboratories, North Chicago, IL), and cefaclor extended-release tablets 375 mg or 500 mg every 12 hours (Ceclor CD; Eli Lilly Pharmaceuticals, Indianapolis, IN). Of these agents, only amoxicillin clavulanate potassium powder for oral suspension and amoxicillin-clavulanate potassium extended-release tablets were designed to treat infections caused by penicillin-resistant pneumococci (penicillin MIC < or =2 microg/mL). Extended release clarithromycin does not provide higher daily doses than its immediate release counterpart; rather, it allows for once-daily dosing of this agent because of its slower absorption following oral administration. Extended-release cefaclor is considered clinically equivalent to 250 mg of immediate-release cefaclor pulvules administered 3 times daily; it cannot be used interchangeably with 500 mg 3-times-daily dosages of other cefaclor formulations. Thus, despite providing a similar or higher total daily dose than its immediate-release counterpart, extended-release cefaclor is indicated only for the treatment of patients with mild to moderate infections caused by susceptible strains of certain organisms. PMID- 15360094 TI - Treatment of acute bacterial rhinosinusitis caused by antimicrobial-resistant Streptococcus pneumoniae. AB - Acute bacterial rhinosinusitis (ABRS) is a secondary bacterial infection of the nose and paranasal sinuses, usually preceded by a viral upper respiratory infection or allergy, with symptoms that have not improved after 10 days or that have worsened after 5 to 7 days. Streptococcus pneumoniae and Haemophilus influenzae are the most common causes of ABRS in adults. Increasing rates of antimicrobial resistance among S. pneumoniae and beta-lactamase production among H. influenzae are formidable challenges to the successful treatment of infections caused by these organisms. To this end, various formulations of amoxicillin clavulanate have been developed, the most recent of which is pharmacokinetically enhanced and provides a total daily dose of 4,000 mg of amoxicillin and 250 mg of clavulanate. This formulation has been shown to be safe and effective in the treatment of infections caused by penicillin-resistant S. pneumoniae (minimum inhibitory concentration 2 microg/mL); the clavulanate component provides adequate coverage of beta-lactamase-producing pathogens. PMID- 15360095 TI - Acute bacterial rhinosinusitis: clinical impact of resistance and susceptibility. AB - Sinusitis is a common disorder associated with notable direct and indirect economic costs. Acute bacterial rhinosinusitis (ABRS) is a relatively poorly defined clinical syndrome characterized by a high spontaneous resolution rate, wide variations in presenting symptoms, and an incomplete understanding of the pathogenesis and clinical course of the disease. Streptococcus pneumoniae and Haemophilus influenzae are the most common causative pathogens in adult ABRS. A relative lack of bacteriological eradication data compared with other respiratory illnesses, uncertainty on the part of many clinicians as to when to treat, and increasing rates of antimicrobial resistance hamper logical treatment strategies. Because it is impossible to know which cases of ABRS will spontaneously resolve and which will not, antimicrobials are recommended. In general, antimicrobial treatment for ABRS should cover both S. pneumoniae and H. influenzae while considering the risk of infection with resistant organisms. Treatment guidelines for ABRS were developed by the Sinus and Allergy Health Partnership in 2000 and were updated in 2004. This article discusses a Sinusitis Therapeutic Outcome Model, a data-driven model used in the development of the treatment guidelines, with respect to different scenarios involving ABRS to illustrate the implications of antimicrobial selection on therapeutic outcome. PMID- 15360096 TI - Streptococcus pneumoniae and community-acquired pneumonia: a cause for concern. AB - Community-acquired pneumonia (CAP) is the sixth most common cause of death in the United States and the leading cause of death from infectious diseases. It is associated with significant morbidity and mortality, and poses a major economic burden to the healthcare system. Streptococcus pneumoniae is the leading cause of CAP. Other common bacterial causes include Haemophilus influenzae as well as atypical bacteria (Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella species). Increasing resistance to a variety of antimicrobial agents has been documented in S. pneumoniae and is common in H. influenzae as well. Successful empiric therapy is paramount to the management of CAP to avoid treatment failure and subsequent associated costs. Given that resistance is increasing among respiratory pathogens, and S. pneumoniae is the most common etiologic agent identified in CAP, strategies for antimicrobial therapy should be based on the likely causative pathogen, the presence of risk factors for infection with resistant bacteria, and local resistance patterns. PMID- 15360097 TI - Review of treatment guidelines for community-acquired pneumonia. AB - Community-acquired pneumonia (CAP) is a costly disease that is associated with significant morbidity and mortality. The growing prevalence of this disease has resulted in various advances in diagnosis and treatment. The most common pathogens of CAP include Streptococcus pneumoniae, Haemophilus influenzae, and atypical pathogens; however, the underlying pathogen often is unknown. Treatment of CAP has evolved because of changing etiologic patterns and increasing antimicrobial resistance among common respiratory pathogens. Among the groups that have established treatment guidelines for CAP are the American Thoracic Society (ATS), the Infectious Diseases Society of America (IDSA), and the Centers for Disease Control and Prevention (CDC). These guidelines establish risk factors associated with drug resistance or infection with specific pathogens. In addition, each guideline provides unique recommendations that are similar in some ways, yet different in others. By understanding the various risk factors for drug resistance and the treatment options endorsed by these guidelines, physicians can treat patients with the most appropriate antimicrobial available. PMID- 15360098 TI - Remote computing environment compensating for brain shift. AB - OBJECTIVE: Anatomical and functional image data become invalid during an operation due to brain shift. Compensation is achieved by using intraoperative imaging to update anatomical information. To accelerate the registration and visualization of pre- and intraoperative image data, the presented work focuses on remote computing capabilities. The underlying framework efficiently combines local desktop computers and remote high-end graphics workstations exploiting expensive hardware. METHODS: By performing all computations on the remote computer, the MR volumes are rigidly aligned via voxel-based registration. Using graphics hardware for acceleration, all interpolation operations are performed with 3D texture-mapping hardware. A new approach then transforms functional markers from preoperative measurements to the intraoperative situation using an automatic tracking algorithm to identify corresponding sulci. Communicating Java viewers are suggested for analyzing the results interactively on a local computer, with all calculations being performed exclusively on the remote computer. RESULTS: The suggested approach was successfully applied in 5 cases using MR data containing functional markers of MEG and fMRI measurements identifying eloquent brain areas. Remote large-scale graphics hardware was thereby efficiently made available for fast registration and interactive direct volume rendering in neurosurgery. CONCLUSION: Overall, the presented framework demonstrates efficient access of expensive high-end hardware remotely controlled by thin clients, and further emphasizes the need to compensate for brain shift in functional neuronavigation. PMID- 15360099 TI - An MRI-compatible surgical robot for precise radiological interventions. AB - OBJECTIVE: A variety of medical robots have been developed in recent years. MRI, including MR angiography and morphological imaging, with its excellent soft tissue contrast is attractive for the development of interventional MRI-guided therapies and operations. This paper presents a telerobotic device for use in CT- and/or MR-guided radiological interventions. A robotic device for precise needle insertion during MR-guided therapy of spinal diseases will be briefly described. MATERIALS AND METHODS: Actuation of robots in an MRI environment is difficult due to the presence of strong magnetic fields. Therefore, the robot was constructed of nonmagnetic materials. The system frame was built from polyether ether ketone (PEEK) and fiber-reinforced epoxy, and actuated using ultrasonic and pneumatic motors. Completely MR-compatible sensors were developed for positioning control. RESULTS: Accuracy evaluation procedures and phantom tests were performed, with the required accuracy of approximately 1 mm being achieved and no significant artifacts being caused by the robotic device during MR image acquisition. PMID- 15360100 TI - Computer assisted pelvic surgery: registration based on a modified external fixator. AB - A fundamental step in Computer Assisted Surgery (CAS) is the registration, when the preoperative virtual data and the corresponding operative anatomy of the region of interest are merged. To provide exact landmarks for anatomical registration, a tubular external fixator was modified. Two intact pelvic bones (one artificial foam pelvis and one cadaver specimen) were used for the experimental setup. Registration was carried out using a standardized protocol for anatomy-based registration in the control group; anatomical registration was achieved using a modified external fixator in the study group. This external fixator had titanium fiducials wedged into the fixator carbon tubes serving as landmarks for paired-point registration. The tubes were used for surface registration. The standard anterior pelvis fixator assembly was augmented with additional bilateral tubes oriented towards the posterior, enabling registration of the sacroiliac areas. The accuracy of registration was checked by "reversed verification", where the examiner used only the screen display to control the virtual position of the pointer tip in relation to selected landmarks. By virtual matching, the real distance was measured with a digital caliper. We defined the verification as "accurate" when the residual distance was less than 1 mm; "acceptable" when it was between 1 mm and 2 mm; and "insufficient" when it exceeded 2 mm. The paired T-test with significance levels of p < 0.05 was used for statistical analysis. The anatomical registration based on the external fixator landmarks was statistically as accurate as that obtained using anatomical landmarks on the pelvic bone. This study concludes that the external fixator, a conventional tool in the management of acute traumatic pelvic instability, can also be useful for landmark registration in CAS. PMID- 15360101 TI - Accuracy of computer assisted percutaneous placement of iliosacral screws: a cadaveric study. AB - OBJECTIVE: To determine the accuracy of computer-assisted fluoronavigation for percutaneous iliosacral screw placement. MATERIALS AND METHODS: A fluoronavigation system (Stryker Navigation System, Stryker Leibinger, Kalamazoo, MI) was used to guide the placement of four iliosacral screws into the S-1 bodies of each of five cadaveric pelvic specimens with intact soft tissues. Accuracy of screw placement was verified by radiographs, CT scans, and direct dissection. RESULTS: All 20 screws were placed accurately without complications. Nineteen screws were completely contained within the osseous "safe zone." On direct dissection, one screw was noted to have penetrated the S-1 foramina by 3 mm without impingement on the nerve root. This was not detected on radiograph or CT scan. CONCLUSION: The results of this study support the safety and accuracy of computer-assisted fluoronavigation for iliosacral screw placement. The advantages include decreased fluoroscopic time, real-time simultaneous visualization of all three views (inlet, outlet, and lateral), and increased accuracy of placement. Clinical study is warranted. PMID- 15360102 TI - Evaluation of neuronavigation in lesional and non-lesional epilepsy surgery. AB - OBJECTIVE: For many patients, surgery for intractable epilepsy provides not only freedom or substantial relief from seizures, but also functional improvement and increased quality of life. Precise intraoperative localization of the underlying structural and functional processes is crucial in this regard. The aim of this study was to clinically evaluate whether neuronavigation leads to an improvement in the precision and safety of epilepsy surgery. In this paper, we also attempt to assess the navigation workstation as a platform for the integration of multimodal information (multimodal information guidance). PATIENTS: Out of a series of 223 epilepsy surgery procedures, 140 were performed with the aid of neuronavigation. Patient and surgical data were prospectively collected. METHODS: We used the neuronavigation device as a common platform to merge complementary information modalities. Correlation of anatomic and structural details with functional information contributed to the surgical script in non-lesional and localization-related epilepsy surgery. At least two different information modalities contributed to planning and surgical guidance in every patient. Immediately following the operative procedure, the surgeon answered a set of questions on the reasons for the application of neuronavigation, and the efficiency and safety of navigation. Detailed analysis of the location of the operative procedure, histopathological findings and outcome was performed. RESULTS: The main benefits of neuronavigation in epilepsy surgery were precision of targeting even in small and deep-seated targets, safe manipulation in critical brain areas, accurate placement of electrodes, and correlation of electro clinical information modalities with underlying structures. Furthermore, navigation provided individual tailoring of craniotomy and corticotomy. It was less reliable for verification of resection boundaries in the case of underlying glioma. Neuronavigational localization and its combination with image fusion and functional investigations greatly improved discussion within the epilepsy surgery team. CONCLUSION: The neuronavigation concept proved its value in epilepsy surgery by linking anatomic, pathologic and functional data of the individual patient. Enhanced by the integration of multimodal information, neuronavigation significantly improved the available treatment options. PMID- 15360103 TI - [Oral leukoplakia. A retrospective study of clinical and histological data]. AB - In this retrospective study, the charts of 91 patients who presented with oral leukoplakia at the Department of Oral Surgery and Stomatology at the University of Berne from August 1995 to December 2001 were analyzed. Critical points in the evaluation included the distribution of age and gender of the patients, type and localisation of the oral leukoplakias, grade of dysplasia, presence and type of potential noxious agents, type of surgical/nonsurgical therapy, and follow-up. The 91 patients included in this study had a total of 145 leukoplakias. Gender distribution corresponded to 39 women and 52 men. Age distribution showed a marked peak in the range of 50-59 years. Sixty-six patients were smokers and 44 patients drank alcoholic beverages on a daily basis. The most frequent localisation of oral leukoplakias was the buccal mucosa (24.5%) followed by the attached gingiva in the mandible including the so-called Trigonum retromolare (17.4%) and the floor of the mouth and ventral tongue (16.8%). Thirty-two of the 52 histologically examined lesions had no epithelial dysplasia, while 14 of the lesions only had a mild and three of the lesions a moderate dysplasia. Of the biopsies taken, three were histologically confirmed to be a carcinoma in situ. PMID- 15360104 TI - [Dose exposure from analog and digital full mouth radiography and panoramic radiography]. AB - The present study investigated the dose exposition in the head and neck region by use of analog and digital radiographic dental systems. Four radiographic devices were tested: panoramic radiography (analog and digital) and 14-image full-mouth survey (FMS, analog and digital). Organ doses were measured on a Rando-phantom by use of CaF2-dosimeters according to the IRCP-guidelines and the effective doses calculated. The results were the following: the value of E was lowest in digital FMS (41 microSv) and highest in analog FMS (78 microSv), i.e. dose was reduced by 47% by using a digital device. In panoramic radiography, doses were 17% lower using digital technique (digital 45 microSv vs. analog 54 pSv). Thus, FMS is no longer associated with higher doses than orthopantomography when conventional films are replaced by digital techniques. PMID- 15360105 TI - [Six simple aids for anxious patients]. AB - This investigation aims at finding a scientifically based method to prevent the traumatization of children in dental treatment. Modern brain research shows that certain simple aids directly interact with the neurological fear responses of the body and memory. With these aids we have been attempting to diminish fear before treatment and to prevent relapse into fear attacks during the treatment. The procedure was surprisingly successful. More than that, it worked equally well with older fear patients. These aids generally raise the treatment to an emotionally comfortable level and lead to better patient cooperation. They put more confidence in the dentist and better relations with the patient.. PMID- 15360106 TI - Five technologies to lead the biotek. PMID- 15360107 TI - Biopharmaceutical and pharmacodynamic studies on topically applied diclofenac gel available in Indian market. AB - In the present investigation, an attempt was made to study and compare, analgesic and anti-inflammatory activity produced by four marketed topical diclofenac formulations in order to justify their usefulness in the treatment of pain and inflammation. By using a diffusion cell, in vitro percutaneous permeation studies were carried out to correlate in vivo activity. The in vivo analgesic activity study was performed by tail flick method on Wistar albino rats. The anti inflammatory activity was performed on rats by carrageenan induced inflammation. It was evident from the study that three among tested three gels; diclofenac permeated effectively through the skin and was able to elicit analgesic and anti inflammatory responses. The study also indicated the presence of therapeutic inequivalence among the marketed topical formulation and the need of bio equivalency and therapeutic equivalency testing of marketed topical applications meant for therapeutic use. PMID- 15360108 TI - Synthesis and properties of N-methyl and N-phenyl derivatives of 1,4-dioxo 1,2,3,4-tetrahydropyrido[3,4-d]pyridazines. AB - 2-Methoxy- and 2-ethoxy-6-methyl-3,4-pyridinedicarboximides (XI, XII) reacted with N-methylhydrazine giving 2- and 3-methyl derivatives of the appropriate 1,4 dioxo-1,2,3,4-tetrahydropyrido[3,4-d]pyridazines (XV, XIII and XVI, XIV). In both cases 3-methyl isomer (XIII, XIV) was formed in higher yield than 2-methyl derivative (XV, XVI). Reaction of the imide XII with N-phenylhydrazine gave the salts of the suitable N-phenyldihydropyrido[3,4d]pyridazines with NH2-NHC6H5 (XXI and XXII) which transformed into N-phenylaminoimide (XXIII) during the boiling in 80% CH3COOH. Imide XXIII isomerized to the appropriate 2-phenyl and 3 phenylpyrido[3,4-d]pyridazines (XXIV - main reaction product and XXV) under the influence of heating in ethanolic solution of C2H5ONa. Some of N phenylpiperazinylhydroxyalkyl(alkyl) derivatives of compound XXIV (XXVII, XXVIII) were pharmacologically active. PMID- 15360109 TI - Ethics in oncology nursing: realism and resources. PMID- 15360110 TI - Returning to the classroom yields new questions about ethics. PMID- 15360111 TI - Are patient safety efforts making a difference? PMID- 15360112 TI - Cardiovascular embryology. AB - During the first 20 days of development, the human embryo has no cardiovascular structure. Over the next month, the heart and great vessels complete their development and look very much like they will at full gestation. This amazing process transforms isolated angiogenic cell islets into a complex, four-chambered structure. During this transformation, the single heart tube begins to beat at 23 days of development and by 30 days blood circulates through the embryo. PMID- 15360113 TI - Fetal cardiovascular physiology. AB - The cardiovascular system of the fetus is physiologically different than the adult, mature system. Unique characteristics of the myocardium and specific channels of blood flow differentitate the physiology of the fetus from the newborn. Conditions of increased preload and afterload in the fetus, such as sacrococcygeal teratoma and twin-twin transfusion syndrome, result in unique and complex pathophysiological states. Echocardiography has improved our understanding of human fetal cadiovasvular physiology in the normal and diseased states, and has expanded our capability to more effectively treat these disease processes. PMID- 15360114 TI - Indications for fetal echocardiography. AB - Congenital heart disease is one of the most common congenital malformations diagnosed in liveborns. As more women undergo prenatal diagnosis, the need for screening fetal echocadiography increases. The fetal, maternal, and familial indications for fetal echocadiography are outlined in order to improve the identification of women in greatest need for this screening modality. PMID- 15360115 TI - Technique of fetal echocardiography. AB - The fetal heart is examined by a series of sequential ultrasound views, which image the cardiac connections and intracardiac structure. The imformation obtained by cross-sectional imaging can be augmented by the use of cardiac measurements, and evaluation with pulsed or color flow Doppler. A thorough familiarity with the normal appearance and systematic evaluation of standard views can exclude or diagnose major heart malformations. With training, cardiac evaluation can be performed during routine obstetric scanning, in a matter of minutes. An accurate description of any abnormal findings and a knowledge of the types of cardiac malformation which are possible, will help the examiner to reach a correct diagnosis. PMID- 15360116 TI - Cardiac arrhythmias in the human fetus. AB - Fetal cardiac arrhythmias have been recognized with increasing frequency during the past several years. Most fetal arrythmias are intermittent extrasystoles, often presenting as irregular pauses of rhythm. These are significant only when they occur with appropriate timing to initiate sustained tachycardia, mediated by anatomic bypass pathways. The most common important fetal arrhythmias are: 1) supraventricular tachycardias, and 2) severe bradyarrhythmias, associated with complete heart block. Symptomatic fetal tachycardias are usually supraventricular in origin, and may be associated with the developmet of hydrops fetalis. These patients may respond to antiarrhythmic drug therapy, administered via maternal ingestion or via direct fetal injection. Such therapy should be offered with careful fetal and maternal monitoring, and must be based on a logical, sequential analysis of the electrical mechanism underlying the arrhythmia, and an appreciation of the pharmacology and pharmacokinetics of the maternal, placental fetal system. Bradycardia from complete heart block may either be associated with complex congential heart malformations involving the atrioventricular junction of the heart, or may present in fetuses with normal cardiac structure, in mothers with autoimmune conditions associated with high titres of anti-SS-A or anti-SS-B antibody, which cross the placenta to cause immune-related inflammatory damage to the fetal atroventricular node. This paper reviews experience with the analysis of fetal caridac rhythm, a detailed discussion of the pathophysiology of arrhythmias and their effect on the fetal circulatory system, and offers a logical framework for the construction of treatment algorithms for fetuses at risk for circulatory compromise from fetal arrhythmias. PMID- 15360118 TI - Guidelines for the evaluation of heart failure in the fetus with or without hydrops. AB - Fetal echocardiography has progressed to be able to diagnose many forms of congenital heart disease and to assess the prognosis of cardiac lesions based on their anatomy and presentation in utero. However, the presence of signs of fetal heart failure, such as hydrops or valvular regurgitation, makes the assessment of prognosis difficult. This article outlines a straightforward method for the rapid evaluation of the fetus who may have congestive heart failure. The differentiation of the prehydropic state from normal is illustrated. PMID- 15360117 TI - Fetal and neonatal cardiac tumors. AB - Primary tumors of the heart are uncommon in the fetus and neonate. Nevertheless, the widespread use of new imaging techniques has contributed significantly to earlier diagnosis, treatment, and thus improved survival. The clinical findings, imaging studies, pathology, and outcome of 224 fetuses and neonates with cardiac tumors collected from the literature are evaluated and discussed. Most tumors are benign, and of these rhabdomyoma is the most common, followed by teratoma, fibroma, oncocytic cardiomyopathy, vascular tumors, and myxoma. Malignant and metastatic tumors are described but are rare. Murmurs, arrhythmias, cyanosis, respiratory distress, and cardiac failure are the main presenting signs of cardiac tumors in the perinatal period. Disturbances in hemodynamic function are correlated with the size and location of the tumor. Cardiac vascular tumors have the best outcome, whereas malignant tumors have the worst. The purpose of this review is to concentrate on the fetus and neonate in an attempt to determine the various ways cardiac tumors differ clinically and morphologically in this age group from those occurring in older children and adults and to show that certain types of tumors have a better prognosis than others. PMID- 15360119 TI - Evolution of heart disease in utero. AB - Cardiac embyogenesis occurs in the first 6 to 7 weeks of human development. Although it is during this time that many of the major cardiovascular defects develop, many of these lesions continue to evolve and others develop in the latter half of gestation. There may be development or progression of ventricular inflow or outflow tract and arch obstruction, and ventricular or great artery hypoplasia. There may be progressive antrioventricular or semi-lunar valve regurgitation which can compromise the fetal circulation. There may be development of dysrhythmias, primary myocardial disease and heart failure. The fetal shunts, the foramen ovale and ductus arteriosus, may change in form and function. Finally, cardiac tumors may develop, grow, or regress. Knowledge of the mechanisms of and potential for progression in fetal heart disease is critical for counseling regarding prognosis and for planning of prenatal and neonatal management. PMID- 15360120 TI - The impact of fetal echocardiography. AB - Fetal echocardiography has impacted the fetus with congenital heart disease in many important ways. Advances in fetal echocardiography have allowed for more accurate and earlier detection of cardiac abnormalities. In turn, the prenatal diagnosis of cardiac abnormalities has improved the care and outcome of selected fetuses with severe cardiac malformations or arrhythmias. Fetal echocardiography has improved the understanding of the development and evolution of congenital heart disease in utero, and it may serve a role in identifying candidates for prenatal intervention. The prenatal diagnosis of congenital heart disease has allowed for better counseling and preparation of families regarding the anticipated prenatal development of the fetus as well as the expected postnatal management plans and prognosis. This article reviews the impact of fetal echocardiography in these and other areas. PMID- 15360121 TI - Advances in fetal cardiac imaging. AB - During the past 25 years, two-dimensional imaging of the fetal heart has evolved into a sophisticated and widely practiced clinical tool, but most heart disease still goes undetected until sometime after birth, despite routine fetal ultrasound evaluations. Over the next 25 years, tremendous advances in fetal cardiac imaging, including three-dimensional imaging, promise to revolutionize both the prenatal detection and diagnosis of congenital heart disease. Image resolution continues to improve year after year, allowing earlier (10-15 week) visualization of the fetal heart, as well as the detection of subtle valvar abnormalities that may progress to serious forms of ventricular hypoplasia at term. However, fetal cardiac imaging remains constrained by limited sonographic windows. To improve the prenatal detection of congenital heart disease, outflow tracts are increasingly included along with the routine screening four-chamber view. However, while the four-chamber view resides within a single plane, lending itself to tomographic evaluation with two-dimensional ultrasound, the outflow tracts (and most forms of congenital heart disease) do not reside within a single plane. For these and other reasons, three-dimensional imaging of the fetal heart ultimately may improve the detection of outflow tract abnormalities, and facilitate comprehension of complex forms of congenital heart disease. Finally, other imaging modalities, including but not limited to Doppler tissue imaging and magnetic resonance imaging, continue to evolve and to complement two- and three dimensional sonographic imaging of the fetal heart. As a result of these ongoing advances in the prenatal detection and assessment of congenital heart disease, these are exciting and glorious times for the field of fetal cardiac imaging. PMID- 15360122 TI - Speech content analysis in feigned depression. AB - The present study investigated the utility of employing computerized speech content analysis software to detect feigned depression in psychologically healthy adults. Twenty-two subjects between the ages of 19 and 54 years old, who scored lower than 10 on the Beck depression inventory (BDI. 1996), were asked to speak freely in response to a neutral stimulus. Participants performed this task in pseudo-randomized order for two experimental conditions: a control state and feigned depression. Computerized transcript analysis data were then subjected to paired t-tests to detect differences based on participants' lexical choices under the two conditions. Results indicated that one semantic category, tool, was significantly different between the feigned depression and control conditions. Another semantic category of interest, arousal, demonstrated a strong trend toward significance. Computerized lexical choice categories previously found to accurately identify persons with depression were not found to be significantly different in the discourse of our participants in the feigning condition. These results are discussed in terms of the use of computerized speech content analysis as a method for possibly identifying persons who may be intentionally feigning depression. PMID- 15360123 TI - The pronoun attraction effect for d(iscourse)-linked phrases: evidence from speakers of a null subject language. AB - Frazier and Clifton (2002) argue that a d(iscourse)-linked wh-phrase such as which boy attracts the reference of a pronoun in a subordinate clause. We translated Frazier and Clifton's materials from English into Romanian. Romanian is a pro-drop language in which null subjects are licensed by person and number agreement on the verb. We found that the d-linking attraction effect held for both pro and overt pronouns in Romanian. The fact that the effect is found for pro provides evidence that the attraction effect is not due to gender matching between the pronoun and the head of the d-linked phrase. We also tested native speakers of Romanian learning English as a second language on Frazier and Clifton's English materials Levels of coreference were highly similar to those for English native speakers and intermediate and advanced learners showed the d linking attraction effect. We discuss the results in the context of Carminati's (2002) Position of Antecedent Hypothesis, arguing that this hypothesis can account for both the fact that higher levels of coreference with a wh-phrase antecedent were found for pro than for an overt pronoun in Romanian and the fact that the coreference levels between an overt pronoun and the wh-phrase antecedent were not elevated for Romanian-speaking second language learners of English. PMID- 15360124 TI - Conceptual locations and pronominal reference in American Sign Language. AB - We report the results of an experiment investigating the ramifications of using space to express coreference in American Sign Language (ASL). Nominals in ASL can be associated with locations in signing space, and pronouns are directed toward those locations to convey coreference. A probe recognition technique was used to investigate the case of "locus doubling" in which a single referent is associated with two distinct spatial locations. The experiment explored whether an ASL pronoun activates both its antecedent referent and the location associated with that referent. An introductory discourse associated a referent (e.g, MOTHER) with two distinct locations (eg., STOREleft, KITCHENright), and a continuation sentence followed that either contained a pronoun referring to the referent in one location or contained no anaphora (the control sentence). Twenty-four deaf participants made lexical decisions to probe signs presented during the continuation sentences. The probe signs were either the referent of the pronoun, the referent-location determined by the pronoun, or the most recently mentioned location (not referenced by the pronoun). The results indicated that response times to referent nouns were faster in the pronoun than in the no-pronoun control condition and that response times to the location signs did not differ across conditions. Thus, the spatial nature of coreference in ASL does not alter the processing mechanism underlying the on-line interpretation of pronouns. Pronouns activate only referent nouns, not spatial location nouns associated with the referent. PMID- 15360125 TI - The acquisition of temporal reference cross-linguistically using two acting-out comprehension tasks. AB - The acquisition of temporal event referencing, encoded by the temporal connectives: then, before, after, when, while, together, until, and since in English, Thai and Lisu was investigated using two acting-out comprehension tasks, a Marble task and a Toy task. Forty children aged 3.6-7.6 years from each language participated. The Marble and Toy tasks differed in their cognitive complexity: in the Marble task the child had to act out only one clause, whereas in the Toy task the child had to act out both clauses. This task manipulation affected performance in Lisu children only. Language-general effects were found, namely "then" and "together" were relatively early and "since" was relatively late in acquisition. Language-specific effects were found for Thai and Lisu. Results confirm that characteristics of task and test sentences affect children's comprehension of sentences expressing temporal relations and can partially account for the disparity in acquisition order found in previous studies. PMID- 15360127 TI - Not just a plasma membrane protein: in cardiac muscle cells alpha-II spectrin also shows a close association with myofibrils. AB - Spectrin and its associated proteins are essential for the integrity of muscle cells and there is increasing evidence for their involvement in signalling pathways as well as having a structural function in mediating stress. Spectrin is a multigene family and it is essential to determine which isoforms are present and their location in the cell. In heart muscle, we have found that one spectrin isoform, alphaII-spectrin, is strongly represented and, using immunofluorescence, we show that it lies within the contractile fibres near the Z-disc as well as on the cardiomyocyte plasma membrane. Electron microscopy of immunogold-labelled cryosections reveals statistically significant clustering of gold particles near the Z-disc, within and close to the edge of myofibrils. betaII-spectrin and ankyrin-R and G are both known to occupy this region. We suggest that alphaIIbetaII spectrin tetramers with ankyrin organise and/or stabilise cardiac muscle cell membrane components relative to the contractile apparatus. PMID- 15360126 TI - Inorganic phosphate affects the pCa-force relationship more than the pCa-ATPase by increasing the rate of dissociation of force generating cross-bridges in skinned fibers from both EDL and soleus muscles of the rat. AB - The effect of inorganic phosphate (Pi) on Ca2+ -activation of actomyosin ATPase activity and force in permeabilized (skinned) single extensor digitorum longus (EDL) and soleus muscle fibers of the rat were investigated. Increasing concentrations of Pi decreased force more than ATPase rate at all Ca2+ concentrations and this effect was more pronounced at submaximal Ca2+ activation. Increasing Pi caused both the normalized pCa-ATPase and pCa-force relationship to be shifted to a higher Ca2+ concentration. At all Ca2+ concentrations ATPase was activated at a lower concentration of Ca2+ than force and this difference in Ca2+ concentration required for the activation of ATPase and force was greater in fast-twitch (EDL) than in slow twitch (soleus) muscle. Soleus muscle pCa-ATPase and pCa-force curves were more sensitive to Ca2+ (pCa50 = 5.97 and 5.89, respectively) than EDL (pCa50 = 5.68 and 5.54, respectively). Finally the shape of the pCa-ATPase and pCa-force curves was similar and not affected by Pi. Analysis shows that Pi increases the rate of dissociation of force generating myosin cross-bridges (ratio of ATPase/force (g(app at all Ca2+ concentration, especially at submaximal Ca2+ -activation levels. Pi effects on g(app) are discussed in terms Pi interacting with the isomerization high force AM*ADP states to form high force transitional AM*ADP*Pi* states which facilitate the dissociation of ADP from AM*ADP. Increasing Ca2+ during Ca2+ -activation of the fibers is associated with a progressive decrease in rate of dissociation of force generating myosin cross-bridges g(app). PMID- 15360128 TI - Dictyostelium discoideum talin A is crucial for myosin II-independent and adhesion-dependent cytokinesis. AB - Cytokinesis in myosin II-null (mhcA-) cells of Dictyostelium discoideum requires adhesion to a substratum. Moreover, such myosin II-independent, adhesion dependent cytokinesis can be divided into two distinct classes: a cell cycle coupled form (cytokinesis B) and a cell cycle-independent form (cytokinesis C). To better understand the mechanisms underlying cytokinesis B and C, we introduced mutations into mhcA- cells and isolated clones that consistently gave rise to large, multinucleated cells. One such clone was found to have a disrupted talA gene, which codes for one of two Dictyostelium homologues of talin. Time-lapse observation revealed that cytokinesis B is partially impaired in mhcA-/talA- cells and that cytokinesis C is completely blocked. MhcA-/talA- cells took significantly longer to complete cytokinesis B than mhcA- cells on regular polystyrene surfaces, but not on a more adhesive surface. During cytokinesis B in mhcA- cells, GFP-TalA localized in the cortex and in dot-like structures on the ventral surface. Like vertebrate talin, these TalA dots may be involved in substrate adhesion and provide the traction needed to efficiently carry out cytokinesis B. During cytokinesis C in mhcA- cells, GFP-TalA was localized in the cortex of future furrow regions, and was highly enriched in cytoplasmic bridges that formed as the furrowing proceeded. Analysis of furrows in which GFP-TalA concentrations were asymmetric suggested that cortical TalA prevents formation of leading edges in regions where its concentration is high and indirectly promotes formation of leading edges elsewhere, which causes passive furrowing of the TalA rich regions. PMID- 15360129 TI - Caldesmon tethers myosin V to actin and facilitates in vitro motility. AB - The current study examines the hypothesis that caldesmon can facilitate the interaction of myosin V with actin filaments by tethering myosin V to actin. Myosin V, purified from bovine brain stem, translocated actin filaments in an in vitro motility assay at a velocity of 0.30+/-0.05 microm/s in the absence of caldesmon at a myosin concentration of 50 microg/ml (ionic strength=110 mM). Filament binding and motility was absent when the myosin concentration applied to the coverslip was reduced to 5 microg/ml, however, the addition of 0.4 microM caldesmon restored binding and motility (0.28+/-0.06 microm/s). This restoration of motility in the presence of caldesmon was blocked by an N-terminal fragment of caldesmon that competitively inhibits the binding of intact caldesmon to myosin. Similar to previous findings with both smooth muscle myosin and platelets (Haeberle et al., 1992; Hemric et al., 1994), these results demonstrate that caldesmon can form a mobile tether that maintains the proximity of myosin V with actin filaments without restricting filament sliding. PMID- 15360130 TI - Myosin heavy chain isoform expression regulates shortening velocity in smooth muscle: studies using an SMB KO mouse line. AB - The kinetics of smooth muscle are thought to be partially determined by the level of the expression of the 7 amino acid insert, SMB, in the myosin heavy chain, as SMB is generally expressed at higher levels in faster smooth muscle. In this study, we determined the role of this insert on shortening velocity and force regeneration following rapid reduction in muscle length (k(step)) in bladder tissue from a transgenic mouse line expressing the insert at three different levels: wild type (WT, +/+, SMB/SMB), an SMA homozygous type (SMB KO, -/-), and a heterozygous type (+/-, SMB/SMA). Smooth muscle from +/+ bladder shorten faster than both the +/- and -/- bladder smooth muscle when activated with Ca2+, consistent with SMB determining the shortening velocity of smooth muscle. The addition of Pi to the fully activated skinned bladder strips did not affect the rate of shortening for either the +/+ or -/- bladder types but did significantly decrease the rate of shortening for the +/- type. In contrast, the addition of ADP to fully Ca2+ activated bladder strips increased the rate of shortening for all three bladder types. However after thiophosphorylation, ADP slowed the shortening velocity. These data are consistent with shortening velocity being determined by the level of activation (or crossbridge attachment) in smooth muscle. The rates of force regeneration according to the k(step) protocol showed no differences between bladder types and also proved insensitive to either Pi or ADP. These data suggest that the rates of force regeneration were determined not only by the kinetics of the crossbridge cycle, but also by factors outside the contractile apparatus. PMID- 15360131 TI - Effects of reduced joint mobility on sarcomere length, collagen fibril arrangement in the endomysium, and hyaluronan in rat soleus muscle. AB - Immobilization is often associated with a decrease in muscle elasticity. This condition is called muscle contracture, but the mechanism is not yet clear. We examined changes in ankle joint mobility, sarcomere length, collagen fibril arrangement in the endomysium, and hyaluronic acid (HYA) in muscular tissue 1, 2, 4, 8, and 12 weeks after immobilization of rat soleus muscles in shortened position. Ankle joint mobility decreased with the duration of immobilization. Sarcomere length had shortened 1 week after immobilization, but did not show further change 2, 4, 8, and 12 weeks after immobilization. Collagen fibril arrangement in the endomysium 1 and 2 weeks after immobilization was longitudinal to the axis of the muscle fibers, whereas 4, 8, and 12 weeks after immobilization it was circumferential. HYA in muscular tissue had increased 1 week after immobilization but remained at the same level at weeks 2, 4, 8, and 12. Histochemically, HYA in the endomysium of immobilized muscular tissue was stained more strongly and widely than that in the control tissue. Increased HYA in muscular tissue may induce muscle stiffness, but the significance of how HYA is related to the mechanism of muscle contracture was not clear. The findings suggest that muscle contracture started 1 week after immobilization and increased with the length of immobilization. Consequently, muscle contracture is affected by the shortening muscle fibers during the early stage of immobilization, after which the collagen adapts by the fibril arrangement in the endomysium becoming more circumferential. This change in collagen fibril arrangement may cause advanced muscle contracture in the late stage of immobilization. PMID- 15360132 TI - Unloaded shortening velocity in single permeabilized vascular smooth muscle cells is independent of microtubule status. AB - Microtubules may influence smooth muscle contraction either via involvement in signal transduction processes or by serving as an internal load that opposes contraction. To test the latter hypothesis, microtubule distribution and the unloaded shortening velocity were investigated in freshly isolated single vascular smooth muscle cells (VSMCs) treated with microtubule modulating drugs. Immunocytochemical studies showed that microtubules run mainly longitudinally in relaxed VSMCs. They are oriented more obliquely, almost transversely to the long axis of the cells after contraction, suggesting that microtubules are compressed during shortening, and thus might impart an internal passive load. Quantitative immunocytochemical analysis revealed that, relative to the control group, colchicine (15 microM) decreased the microtubule density by 40% while taxol (10 microM) increased the microtubule density by 46%. However, alteration of microtubule polymerization status by these microtubule-modulating drugs did not have a significant effect on unloaded shortening velocity in alpha-toxin permeabilized VSMCs under maximal activating conditions or submaximal activating conditions (about 36% of maximal velocity). These results suggest that microtubules do not present an appreciable internal load to dampen single VSMCs shortening in the present experimental system, and that their influence on smooth muscle contraction is primarily via signal transduction mechanisms. PMID- 15360135 TI - [Centenary and bicentenary]. PMID- 15360136 TI - [Pain management, a new culture]. PMID- 15360133 TI - Long-term growth selection of mice changes the intrinsic susceptibility of myogenic cells to apoptosis. AB - Myogenic cells were derived from mice long-term selected over 70 generations for high 6-weeks body weight (DU-6) and from unselected control mice (DU-Ks). The cells were grown in medium with 10% foetal bovine serum (FBS) for 8 days or transferred to low serum conditions (1% FBS) at days 4 and 6 of cultivation, respectively, and maintained for two further days. In both cell lines, serum reduction induced decreases in DNA and protein contents, and in DNA synthesis rate. It also triggered apoptotic cell death as demonstrated by increased DNA strand breaks and expression of active caspase-3. Concomitantly, the anti apoptotic protein bcl-2 was enhanced. The basal frequency of apoptotic cells decreased with time of cultivation in both lines and was lower in DU-6 than in DU Ks cells. However, the increase in apoptosis induced by serum reduction was more pronounced in DU-6 than in DU-Ks cells and did not differ between the time points of serum reduction. The results suggest that growth selection decreases the basal apoptosis frequency of muscle satellite cells under normal supply, but enhances the intrinsic susceptibility to growth factor withdrawal by serum deprivation as a severe apoptotic stimulus. Furthermore, the apoptotic response to growth factor withdrawal seems to be largely independent of the stage of myogenic development. PMID- 15360134 TI - Function induced modifications of gene expression: an alternative approach to gene therapy of Duchenne muscular dystrophy. AB - In Duchenne muscular dystrophy a large gene that codes for dystrophin is altered. The possibility that the defective gene/protein could be at least in part substituted by other molecules that the diseased muscle is able to produce and that have a function similar to that of dystrophin is being discussed. Muscle fibres have a tremendous adaptive potential, and the expression of several protein isoforms can be induced by either stretch or long-term change of activity. The exploitation of this ability of muscle cells to express new genes, which would code for proteins that will not be alien to the individual, for treatment of Duchenne muscular dystrophy is being considered. The argument for this approach is strengthened by results that in patients with Duchenne muscular dystrophy the progress of the disease can be slowed with changes of muscle activity. PMID- 15360137 TI - [The outcome of allergology]. PMID- 15360138 TI - [Emergency medicine, one of the most acute fields in medicine]. PMID- 15360139 TI - [Cancer in 2004]. PMID- 15360140 TI - [Two key words for tomorrow's cardiology]. PMID- 15360141 TI - [Digestive surgery, between the old and the modern]. PMID- 15360142 TI - [Maxillofacial surgery, reconstruction of a balanced face]. PMID- 15360143 TI - [New horizons in dermatology]. PMID- 15360144 TI - [Diabetes, a worldwide disease]. PMID- 15360145 TI - [You said... endocrinology!]. PMID- 15360146 TI - [Epidemiology, a junction point discipline]. PMID- 15360147 TI - [Hepato-gastroenterology, current expectations and future concern]. PMID- 15360148 TI - [From genomics to treatment]. PMID- 15360149 TI - [Geriatrics are now a part of the medical landscape in France]. PMID- 15360150 TI - [What's new in gynaecology-obstetrics?]. PMID- 15360151 TI - [Future vision of haematology]. PMID- 15360153 TI - [Prospects in imaging]. PMID- 15360152 TI - [Regarding infectious diseases]. PMID- 15360154 TI - [Immunological expectations at the dawn of the XXIth century]. PMID- 15360155 TI - [Physiotherapy in motion]. PMID- 15360156 TI - [Internal medicine, deeds and stakes]. PMID- 15360157 TI - [Palliative care: caring for human beings in the context of scientific medicine]. PMID- 15360158 TI - [Neurology in 2004]. PMID- 15360159 TI - [Clinical and preventive nutrition]. PMID- 15360160 TI - [Technology at the service of sight]. PMID- 15360161 TI - [Laryngology and rhinology, a dynamic domain]. PMID- 15360162 TI - [Diagnose, treat and repair hearing disorders]. PMID- 15360163 TI - [Speech therapy, the springboard of the past, energy of the present and the boom of the future]. PMID- 15360164 TI - [Paediatrics, experience, concerns, and expectations]. PMID- 15360165 TI - [Pneumology in 2004, as dynamic as ever]. PMID- 15360166 TI - [Psychiatry is a discipline]. PMID- 15360167 TI - [Rheumatology, prospects of development]. PMID- 15360168 TI - [The third corner stone of a renewed medical practice]. PMID- 15360169 TI - [The nursing profession, what's in store?]. PMID- 15360170 TI - [Reflections on therapeutic progress in daily practice]. PMID- 15360171 TI - [Psychotherapy, between the 19th and 21st centuries]. PMID- 15360172 TI - [Nurses and public health... public health nurses]. PMID- 15360173 TI - [Public health in Quebec at the dawn of the 21st century]. AB - This paper summarises the status of public health in Quebec at the dawn of the 21st century. After introducing the current definition, the author lays out five basic functions of public health in Quebec: knowledge and surveillance of populations health and wellbeing, health protection; prevention of disease, trauma and social problems that influence health; health and wellbeing promotion; and service organization and evaluation. The organization of public health services is then described at the local level (CLSC), regional level (Public health units) and national level (Ministry, Public health directorate and National Institute of Public Health). Finally, the trends and priorities elaborated over the last ten years, as well as the National Public Health Program to be implemented over the next ten years are described. PMID- 15360174 TI - [Nurses and public health: evolution of missions and trainings]. AB - Reflection upon the role of nurses in public health is not something new; however, over the past few years, a questioning and similar reports have sprung up pointing to the difficulty perceived by nurses in reconciling their professional practice and public health. Currently, in light of the evolution of public health, where does the practice of nursing fit into public health practice? What are the potential movements and trends in terms of training? The goal of this paper is to present a number of possible avenues for thought on the subject, namely concerning the training and the allocation of competencies. It would be undoubtedly a pity to conceive of public health training today being intended only for nurses. It is rather preferable that post-graduate public health training is aimed at a variety of professional categories, regardless of their initial training or degree course. The development of schools of public health to which nurses could have access in the same way that other health professionals or from the social and educational fields do is critical. Above and beyond what is at stake for specialisation, and even independent of the implication of nurses in public health work, the current trend to re-balance the work and tasks of both nurses and doctors constitutes a significant challenge for public health. PMID- 15360175 TI - [Challenges facing nurses' associations and unions: a global perspective]. AB - Nurses are at the heart of every country's health care system. What sort of problems do they face at work? What are the coping strategies that they and their organizations pursue? Drawing on the findings of a unique global survey of nurses' associations and unions, the authors provide clear answers to these and related questions at a time when population ageing, epidemics, privatization and understaffing are putting heightened pressures on health care systems around the world. Interestingly, the priority concerns of nurses' organizations the world over have much in common despite the widely differing national contexts in which they operate. PMID- 15360176 TI - [Development of a reference mark of skills to practice nursing in the school setting]. AB - This article describes the different skills which a school nurse can be brought to mobilise while practicing his/her profession. In French speaking Switzerland, these professionals acquire general community health training after receiving their basic training. The specificity of their target population group (students and their families), the context of their workplace (the school and its various actors), their daily interdisciplinary practice all put into question the adequacy of this training. The goal of this work was the formulation of the different skills required in order to conduct well their complex professional activity, in the format of a referential of competencies, prior to the development of a more specific training programme. The referential was built upon the basis of the "professional-group" approach; it was compiled from the analysis of working situations representative of the diversity in practice, by a group of confirmed professionals. PMID- 15360177 TI - [Nursing practice in the context of geographic isolation: implications for training]. AB - A survey on the characteristics of the unique working context of nursing practice in remote areas of French Polynesia and semi-remote areas of northern Quebec demonstrates the importance of a specific training to best prepare the professionals who find themselves in this type of setting. Twenty professionals were interviewed: six nurses practicing in isolated stations in French Polynesia (Tuamotu Archipelago, Marquesas Islands and Austral Islands), six nurses practicing in semi-remote areas within northern Quebec (namely among the Algonquins, the Crees and the Attikameks), four officials of the French Polynesian Health Directorate and four training programme designers from Quebec who were encountered during an expedition to Montreal, Mistissini and Trois Rivieres. The authors identified ten characteristics which were then regrouped into two categories for both of the practice contexts: first, those inherently linked to professional practice in an isolated context (including the characteristics of nursing practice, the working conditions, the community's health problems, their forms of socio-professional relations, their way of life, and their perception and responses to isolation); and second, those pertaining to the social and natural environment, the economic conditions and the community's cultural specificities. All of the results strongly demonstrate that the specificities of the skills utilised by these nurses are indeed very different than those which they received in their initial preparatory training. If a training programme specific to nursing practice in isolated settings seems essential for these nurses, their individual predisposition to practice in such a complex environment and the acknowledgement of their professional competencies are equally to be considered within the perspective of human resources management. PMID- 15360178 TI - [The nursing labour market in Canada (1985-1999)]. AB - This literature review analysed both published and unpublished scientific and professional studies on the nursing labour market in Canada within the period of 1985 to 1999. The goal was to conduct a situational analysis utilising statistical data and canvassing all concerned parties to extract their points of view. The analysis revealed significant cyclical variations in the evolution of the workforce, particularly with respect to auxiliary nurses, such as the perceived existence of major problems in recruiting new professionals in the field and retaining existing professionals in their organisations, the lack of homogeneity in educational training programmes, and the co-existence of several operational structures for organising nursing care, of which there is a lack of evaluation on their effectiveness. The results of the literature review identify the necessity to further develop the knowledge base on such a relevant dimension of the nursing labour market. PMID- 15360179 TI - [Public health nurses in Quebec: an essential contribution that remains to be known]. AB - Public health nurses in Quebec are essential although little is known about their role. Generally trained at the undergraduate level, with additional qualifications in community health, they essentially work in CLSCs in maternal and child health programs, youth clinics or occupational health. In regional public health departments, they also work in the field of infectious disease prevention or other programs. Like their colleagues, they are confronted to the profession's challenges, the most notable being current and future shortage of nurses. However, they can now count on a new. As nurses further develop their practice and orient it even more towards primary prevention and health promotion, they will significantly contribute to the National Public Health Program and to the improvement of population health and wellbeing. PMID- 15360180 TI - [Disease prevention and health promotion for children and youth: current status and contemporary issues in CLSC-based nursing practice]. AB - This article aims to provide an overview of the disease prevention and health promotion work carried out by CLSC nurses working with infants, children and youth. A second objective is to present our synthesis of four recent studies on the state of prevention-promotion in these organizations. This is followed by our analysis of several contemporary issues related to nursing practice in this intervention sector. While disease prevention and health promotion is a major component of the practice of CLSC nurses who work with infants, children and youth, it is an intervention sector that has been weakened by numerous changes affecting these organizations in the wake of the reform. The discussion thus proposes several courses of research and action in the hopes of strengthening this major component of the CLSC mission. PMID- 15360181 TI - [Equal at birth-growing healthy: an example of nurses' involvment in perinatal health promotion and prevention]. PMID- 15360182 TI - [Heart health promotion and preventive practices of Quebec nurses working in local community health centers]. AB - The heart health promotion and prevention practices of nurses working in front line services in 20 Quebec CLSC's are analyzed through a qualitative (n = 47 nurses) and a quantitative approach (n = 177 nurses and n = 3802 patients). The results demonstrate that although nurses state they integrate prevention and promotion in their daily practice, patients report that nurses provide few prevention recommendations. The major challenge for prevention-promotion is to strengthen secondary prevention and, especially, integrate primary prevention and promotion in nurses' daily clinical practice. PMID- 15360183 TI - [Health promotion of the elderly: a conceptual matter]. AB - Although community health nurses have to face the increasing needs in curative and palliative care, they still engage in health promotion activities for the elderly, mainly cognitive and behavioural ones. The Quebec community health nurses who choose the McGill model of nursing, are able to integrate health promotion interventions in their daily practice with old aged people. In fact, this model helps these nurses consider the elderly, health, sickness and nursing care in a way that makes them promote self-esteem, empowerment, development and coping through most of their interventions. Moreover, the elderly population would benefit from an increased implication of nurses in ecological, environmental and community health promotion projects. PMID- 15360184 TI - [The daily public health nursing practice: the Montreal infectious diseases unit as an example]. AB - The health of urban populations depends on multiple factors, among them access to health and social services. The Montreal Public Health Department (PHD) is part of such services, where a number of nurses are working within a multidisciplinary team. It is through the description of the work accomplished by the team for infectious diseases control and prevention, and the vignettes of real life events related to the follow-up of syphilis, tuberculosis and hepatitis A cases, as well as quarantined individual exposed to SARS that the authors want to illustrate the work of the urban public health nurses. The examples are preceded by a description of the socio-demographic characteristics of the city of Montreal, that influence health problems, and therefore nurses' work. PMID- 15360186 TI - [Careers and accounts of nurses]. PMID- 15360185 TI - [Design and implementation of occupational health programs in the workplace: the Quebec approach]. AB - This paper describes the Quebec experience in the design and implementation of occupational health programs in the workplace. To begin with, a brief overview of the historical context and organisational framework of occupational health are presented. Next, each of the phases involved in the design of occupational health programs is described: identification of workers' health and safety problems, selection of action priorities, and program design, implementation and evaluation.... In the end, the roles and responsibilities of the doctors, nurses and industrial hygienists generally involved in these multidisciplinary teams are presented. PMID- 15360187 TI - [Nurse... a professional also responsible for teaching public health. Interview by Laurence Fond-Harmant]. PMID- 15360188 TI - [Training... but moreover, confrontation with the field. Interview by Laurence Fond-Harmant]. PMID- 15360189 TI - [From the hospital to social security insurance... Interview by Laurence Fond Harmant]. PMID- 15360190 TI - [From humanitarian action to health pedagogy. Interview by Laurence Fond Harmant]. PMID- 15360191 TI - [A public health nurse working: a different approach to mankind and health]. PMID- 15360192 TI - [The most essential is the way..., career of a Brazilan nurse]. PMID- 15360193 TI - [Educating the patient: skills needed to nursing at the crossroads of care and pedagogy]. AB - The educational team interrogated itself concerning its pedagogical competence within the framework of its educational activity for asthmatics: how does the care provider become a learning facilitator for the asthmatic person? Among the possible responses, there were two aspects which appeared essential: understanding the patient's learning logic and taking into consideration his/her representations and prior knowledge. The concrete translation of these principles primarily rests upon an educational programme truly centred on the person in question, and one which uses interactive methods, promoting the transfer of knowledge in daily life. Then the caregiver utilises previous experience and the asthmatic's representations in order to solicit a cognitive conflict, a source of learning. PMID- 15360194 TI - [Public health and training for nurses and senior level management]. AB - The teaching of public health has progressively developed within the area of nursing and senior management training, in line with the development of these professions and the population's needs, in order to hold an identifiable place in today's programmes. Its dynamic is closely linked to the educational projects of various institutes. But if training is strengthened to develop this pertinent and significant dimension as an integral part of care providers' functions, then public health as a professional practice remains dependant upon institutional objectives and political and economic trends. PMID- 15360195 TI - [Hospital nurses' perception of public health]. AB - The objective of this work was to study hospital nurses' perceptions of public health based on a pedagogical exercise. A qualitative study, lead by students of a training institute for nursing care (IFSI), was conducted using semi-directed interviews of hospital nurses from different departments of a provincial hospital. The main results show that for one-third of the nurses interviewed, the hospital is not a setting for public health practice. In terms of their definition of public health, it is closely linked to the concept of prevention, with primarily individual approaches. This definition is very close to the public health activities that they conduct at the hospital and which are centred on information and disease prevention, on education related to pathologies and the relationship between the provision of care and listening to the patients and their families. Few nurses place their public health activities within the scope of the areas of hospital cleanliness, the welcoming of the patients, the organisation of services, and the improvement of the quality of care. The potential tracks which have emerged from this work lead to the need for the strengthening of training in patient education, in the hospital's work networking with external partners, and in better development of public health activities undertaken in the hospital setting. PMID- 15360196 TI - [Public health nurses: a profession under construction]. AB - In France, the Training Institutes for Nursing Care (IFSI) primarily prepare individuals for care providing occupations. Thus nurses practicing in the field of public health are non-practicing care providers since their recruitment is solely based upon the criteria of having a State Nursing Diploma (DE) at the end of their initial training. If one admits that public health is scattered and dispersed in France, then the situation for nurses is as much so and on many levels. Assignments only slightly formalised, and a lack of regulation and heterogeneous training compromise the legibility of this profession which nevertheless strongly exists in fact in practice. This remark is notably illustrated in the situation of DDASS and DRASS nurses. State bureaucrats, equipped with their one experience in care provision without any specific supplementary training, they work with the health administrators and the multidisciplinary teams to implement national public health policies. They are few, geographically and operationally isolated, and many of them follow training courses with are often restrictive in nature due to the fact that they take their personal time to do them. These are the main elements of the analysis of position announcements and different work being done within the professional network in the field, issued following a study conducted by the National Association of Public Health Nurses (ANIDESP) in 2002, and further enhanced by the conclusions of a seminar held in March 2003 at the Ministry of Health. The framework of a newly formed reference, missions in abeyance of formalisation and the establishment of a training programme, these are the main supporting points of the different proposals that the authors are making to the Health Directorate, and the Personnel and Budgeting Departments of the Ministry of Health and Social Affairs. PMID- 15360197 TI - Comprehensive survey of combinatorial library synthesis: 2003. PMID- 15360198 TI - Polystyrene-supported phosphine-catalyzed aza-Baylis-Hillman reactions and the relationship between resin loading level and catalyst efficiency. PMID- 15360199 TI - Construction of a 3-amino-2-pyridone library by ring-closing metathesis of alpha amino acrylamide. PMID- 15360200 TI - Method for the parallel preparation of the aspartic protease isostere: hydroxyethylamino amides. PMID- 15360201 TI - Palladium-catalyzed synthesis of aldehydes from aryl iodides and acetic formic anhydride. PMID- 15360203 TI - Combinatorial synthesis of insoluble oxide library from ultrafine/nano particle suspension using a drop-on-demand inkjet delivery system. PMID- 15360202 TI - Simple coupling reaction between amino acids and weakly nucleophilic heteroaromatic amines. PMID- 15360205 TI - Simultaneous in-situ monitoring of parallel polymerization reactions using light scattering; a new tool for high-throughput screening. AB - A recently introduced technique, simultaneous multiple sample light scattering (SMSLS), was used to monitor parallel polymerization reactions in situ. SMSLS is designed for real-time, high-throughput screening and provides a time-dependent light scattering signature for each reaction, which contains both qualitative and semiquantitative information. Qualitatively, the signature immediately indicates whether the reaction occurs or not, whether there is an initial lag period, and how long the reaction takes until it stops. The signature also provides estimates of the reaction rate and weight average molecular mass M(w), and its shape can help identify mechanistic aspects, for example, controlled versus free radical polymerization, presence of impurities, etc. The method is inherently adapted to small sample volumes and requires no special sample preparation or postpolymerization characterization. The demonstration here involved the free radical polymerization of acrylamide under varying conditions and should be readily applicable to a wide variety of other reactions. Results were cross checked with multi-detector gel permeation chromatography. PMID- 15360204 TI - Identification of synthetic phosphatidylserine translocases from a combinatorial library prepared by directed split-and-pool synthesis. AB - Simple sulfonamide and amide derivatives of tris(2-aminoethyl)amine (Tren) are known to promote the translocation or flip-flop of phosphatidylcholine, but not phosphatidylserine, across bilayer membranes. This paper describes the synthesis of a 300-member, spatially encoded library of Tren derivatives with appended peptide--sulfonamide and peptide--urea arms. The library was synthesized using the Encore method with SynPhase lanterns as the solid support. A high-throughput assay was developed to screen individual members of the library for an ability to translocate a fluorescent NBD derivative of phosphatidylserine across vesicle membranes. Several lead compounds were identified, and one was synthesized independently to confirm its high phosphatidylserine translocation activity. PMID- 15360206 TI - Palladium-catalyzed synthesis of uridines on polystyrene-based solid supports. AB - In this paper, the solid-phase synthesis of various substituted pyrimidine nucleosides is described starting from 2'-deoxyuridine, which has been attached through a base labile linker to polystyrene resins. The utility of the Pd(0) cross-coupling to functionalized pyrimidine nucleosides is expanded herein to include reactions of resin-supported 5-iodo-2'-deoxyuridine under Sonogashira, Stille, Heck, and Suzuki conditions. Upon cleavage with MeONa, a library of 5 substituted pyrimidine nucleosides was obtained in good (under Sonogashira and Stille conditions) to moderate (under Heck or Suzuki conditions) yields and high purity. Except the Suzuki-type reactions, the presented methods exhibit a significant improvement and facilitate the synthetic procedure with respect to purification and yields (determined after filtration over silica gel). PMID- 15360207 TI - A solution- and solid-phase approach to tetrahydroquinoline-derived polycyclics having a 10-membered ring. AB - With the goal of library generation using a polycyclic derivative 5 having an enamide functional group, a simple and practical, enantioselective synthesis of tetrahydroquinoline derivative 2 was achieved. The phenolic hydroxyl group in compound 2 was utilized as an anchoring site for solid-phase synthesis. The ring closing metathesis approach yielded the desired polycyclic product 5 on solid phase in five steps (overall 40% yield). Compound 5 is a novel scaffold for the library generation of natural product-like polycyclics having a functionalized medium ring for obtaining a new class of small molecules to be utilized as chemical probes. PMID- 15360209 TI - Parallel solid-phase syntheses of 1,3,4-thiadiazolium-2-aminides. AB - An efficient and advantageous solid-phase strategy has been developed to synthesize 1,3,4-thiadiazolium-2-aminides. The title compounds were prepared in parallel fashion according to the following compact route: (i) anchoring of aromatic aldehydes to the solid support; (ii) solution preparation of 1,4 disubstituted thiosemicarbazides from hydrazines plus isothiocyanates; (iii) trimethylsilyl chloride-promoted cyclization between the resin-bound aldehydes and 1,4-disubstituted thiosemicarbazides; and (iv) removal of the products from the solid support by acid treatment. The products (17 made in all) were cleaved with high initial purities (90-98%) and obtained in generally good isolated yields (53-94%, with one exception). PMID- 15360208 TI - Solution- and solid-phase synthesis of natural product-like tetrahydroquinoline based polycyclics having a medium size ring. AB - A solid-phase synthesis of tetrahydroquinoline-derived polycyclic 4, having a medium size ring with an enamide functionality, was achieved from tetrahydroquinoline derivative 3 in five steps with overall 40-45% yield. An enantiopure, tetrahydroquinoline-derived beta-amino ester, 1, was converted into compound 2 that has a free phenolic hydroxyl group as an anchoring site for solid phase synthesis. The solid-phase worked well for this sequence, in which the synthesis of the unsaturated eight-membered enamide lactam was obtained by a ring closing metathesis approach. Compound 4 is a novel, natural product-like polycyclic derivative that could further be utilized in library generation for developing small molecule chemical probes. PMID- 15360212 TI - New synthetic pathway to diverse 2-substituted quinolines based on a multicomponent reaction: solution-phase and solid-phase applications. AB - Using Kobayashi's modification of the Grieco reaction, we were able to synthesize diverse 4-phenylthio-1,2,3,4-tetrahydroquinolines. These intermediates were oxidized and subsequently pyrolized to provide the corresponding quinolines. This new approach to 2-substituted quinolines was exemplified by liquid-phase production of a 25-member library. This was extended to solid-phase chemistry, starting from (l)-4-nitrophenylalanine on Wang resin, for production of a 16 member library. The latter compounds possess potentially interesting VLA-4 antagonist properties. PMID- 15360211 TI - A new class of arylpiperazine derivatives: the library synthesis on SynPhase lanterns and biological evaluation on serotonin 5-HT(1A) and 5-HT(2A) receptors. AB - An efficient solid-supported method for the synthesis of a new class of arylpiperazine derivatives containing amino acid residues has been developed. A 72-membered library was synthesized on SynPhase Lanterns functionalized by a BAL linker. A one-pot cleavage/cyclization step of aspartic and glutamic acid derivatives yielded succinimide- and pyroglutamyl-containing ligands (chemsets 9 and 10). The library representatives under study showed different levels of affinity for 5-HT(1A) and 5-HT(2A) receptors (estimated K(i) = 24-4000 and 1-2130 nM, respectively). Several dual 5-HT(1A)/5-HT(2A) ligands were found, of which two (9(3,3) and 9(3,5)) displayed high 5-HT(2A) affinity comparable to that of the reference drug ritanserin. A set of individual fragment contributions for the prediction of 5-HT(1A) and 5-HT(2A) affinity of all the library members were defined on the basis of the Free-Wilson analysis of 26 compounds. An alkylarylpiperazine fragment had essentially the same impact on the affinity for both receptors, whereas different terminal amide fragments were preferred by 5 HT(1A) (chemset 17, R(2) = adamantyl) and 5-HT(2A) (chemset 9, R(2) = norborn-2 ylmethyl) binding sites. PMID- 15360210 TI - DNA transfection screening from single beads. AB - The solid-phase synthesis of a library of arginine-containing lipid transfection agents on high-loading beads is described. The transfection activity of the cationic lipids was determined using compound cleaved from single beads (single bead screening) and showed, in some cases, comparable or higher DNA transfection activities as compared to commercially available reagents. Lipids with one arginine headgroup and a cholesterol tail were found to be the most active, even though their DNA binding strength (retardation assays) was relatively weak. Single-bead screening of transfection activity facilitates the rapid analysis of libraries of transfection reagents and will allow the rapid optimization of gene delivery into cells, both in culture and in vivo. PMID- 15360213 TI - An expedient and facile one-step synthesis of a biguanide library by microwave irradiation coupled with simple product filtration. Inhibitors of dihydrofolate reductase. AB - It has been demonstrated previously by us that guanide-containing compounds (1 and 2) can inhibit significantly dihydrofolate reductase (DHFR). In this report, we have produced an array of alkyl- and aryl-based biguanide compounds using microwave irradiation. Further, we have demonstrated the use of TMSCl for the first time as an excellent and practical catalyst for the formation of alkyl and aryl biguanides. Using these methods, we prepared a 60-compound collection, of which one compound (21g) showed approximately one-half of the inhibitory activity of the parent compound 2. PMID- 15360215 TI - Solid-phase synthesis of an alkylaminobenzanilide library. AB - The synthesis of a library of 2- and 3-substituted benzanilides has been achieved on solid phase. Attachment of anilines to formyldimethoxyphenyl (FDMP) resin via reductive amination was optimized to allow a wide range of anilines to be used. Acylation of this resin-bound aniline was accomplished with 2- or 3-nitrobenzoyl chloride to yield nitrobenzanilides. Following reduction of the nitro group, the resulting amine was alkylated using aromatic and heteroaromatic aldehydes in the presence of NaBH(OAc)(3) under controlled conditions. Finally, the products were cleaved from the resin using trifluoroacetic acid to produce a 10 800-member library. PMID- 15360214 TI - Iron carbonyl-mediated parallel solution-phase synthesis of cyclohexadienoic acid amides. AB - Iron carbonyl-stabilized cations have been employed to develop methodology for carbon-carbon and carbon-heteroatom formation suitable for the preparation of combinatorial libraries. Different nucleophiles were added to tricarbonyl(cyclohexa-1,3-dienylcarboxylic acid 4-nitro-phenyl ester)iron hexafluorophosphate. Aminolysis, followed by decomplexation, yielded substituted cyclohexadienyl amides of high purity. Carbon, oxygen, and sulfur nucleophiles gave good results, while amine nucleophiles gave products of somewhat lower purity. PMID- 15360216 TI - One-minute full-gradient HPLC/UV/ELSD/MS analysis to support high-throughput parallel synthesis. AB - High-throughput parallel synthesis of library compounds for early drug discovery requires high-throughput analytical methods to confirm synthesis, identify reaction products, and determine purity. An ultrafast 1.0-min HPLC/UV/ELSD/MS method was developed and compared to our standard 2.5- and 5.0-min methods in order to determine if the faster method was appropriate to evaluate compound synthesis and determine purity. In addition to using standard test mixtures, a 400-member library produced by high-throughput parallel synthesis was used for comparing the various methods. Mass spectrometric detection was used for compound identification, while UV and ELSD data offered purity assessment. Compared to our longer separations, chromatographic separation achieved using the 1.0-min method was sufficient for compound evaluation and purity assessment. This ultrafast 1.0 min HPLC/UV/ELSD/MS method is expected to increase analytical throughput tremendously, provide important information faster, and reduce the overall cycle time from synthesis to screening. PMID- 15360217 TI - Colorimetric tools for solid-phase organic synthesis. AB - One of the unresolved problems of solid-phase organic synthesis (SPOS) is the availability of general and rapid methods to monitor the transformation of functional groups present in molecules supported on insoluble supports. Color tests, far from providing the ultimate solution, may help in detection (and sometimes in quantification) of different functional groups. In this short review, we have collected most of the methods available and applied in SPOS with an Experimental Section that describes the procedure we have successfully applied to bead analyses in our laboratories. PMID- 15360218 TI - Multistep parallel synthesis of substituted 5-aminobenzimidazoles in solution phase. AB - An efficient solution-phase parallel synthesis of multisubstituted 5 aminobenzimidazoles is described. The two fluorine atoms of 1,5-difluoro-2,4 dinitrobenzene (DFDNB) are sequentially and quantitatively replaced by nucleophiles. Simultaneous reduction of aromatic m-dinitro groups by Pd C/HCOONH(4) results in 2,4,5-benzenetriamines, which are continuously condensed with aldehydes to successfully construct the benzimidazole ring without additional oxidants. The free aromatic amino group is further modified by anhydrides, isocyanates, isothiocyanates, and sulfonyl chlorides. All the reactions involved here are highly effective in giving the desired products at room temperature. Four diversity points are introduced in the final products. PMID- 15360219 TI - Smart cleavage reactions: the synthesis of an array of ureas from polymer-bound carbamates. AB - The solid-phase synthesis of a library of di-, tri-, and tetrasubstituted ureas is described. In this approach, an array of polymer-bound carbamates was synthesized. These polymer-bound primary and secondary amine carbamates were then treated under "smart" diversity-building cleavage conditions using a series of aluminum amide complexes to form the corresponding urea cleavage products. The crude cleavage products from this study were isolated in excellent yield and purity. To show the applicability of this strategy, a series of biaryl ureas were synthesized using sequential solid-phase Suzuki coupling and urea formation reactions. PMID- 15360220 TI - A parallel solution-phase synthesis of substituted 3,7 diazabicyclo[3.3.1]nonanes. AB - The parallel solution-phase synthesis of a series of building blocks and combinatorial libraries based on natural bispidine scaffold has been accomplished. Key reactions include catalytic hydrogenation of the (-)-cytisine heterocyclic system, followed by alkali-mediated ring cleavage. Using this approach, a series of new bispidine core building blocks for combinatorial synthesis with three points of diversity were effectively synthesized. The libraries from libraries were then obtained in good yields and purities using solution-phase acylation reactions. Obtained combinatorial libraries of 3,4,7 trisubstituted bispidines are potentially useful in the discovery of novel physiologically active compounds. PMID- 15360221 TI - Solid-phase convergent synthesis of a benzimidazolone library via the combination of two smaller arrays of carboxylic acids and secondary amines. AB - The concept of convergent synthesis can be extended to combinatorial chemistry in order to obtain collections of products characterized by considerable chemical diversity and a certain molecular complexity. In this work, a library consisting of three carboxylic acids containing a benzimidazolonic functionality with variations at two positions was synthesized on solid phase. After cleavage, this library was combined with a second library consisting of 16 solid-supported amines containing two points of variation. IRORI technology was used for the split-and-mix synthesis of the final 48 members library. PMID- 15360222 TI - Resin-bound 4H-1,3-oxazine-masked beta-diketones for functionalizing cleavage strategy. AB - Resin-bound 4H-1,3-oxazines are synthesized by the stepwise condensation of an amide resin, an aldehyde, and an alkyne. Upon DDQ activation, oxazines are converted into oxazinium salts. When treated with hydrazines, these resin-bound beta-diketone equivalents yield pyrazoles through a functionalizing release process. This multicomponent capture strategy, tedious to handle in classical synthesis in solution, is well-suited to solid-supported chemistry. It facilitates the handling of sensitive and unstable intermediates, such as N-alpha methoxyalkylamides and 1,3-oxazinium salts. PMID- 15360223 TI - Copper(I) complexes of fluorinated triazapentadienyl ligands: synthesis and characterization of [N[(C3F7)C(Dipp)N]2]CuL (where L = NCCH3, CNBut, CO; Dipp = 2,6-diisopropylphenyl). AB - Sterically demanding triazapentadiene [N[(C3F7)C(Dipp)N]2] H has been synthesized in good yield. It features a W-shaped ligand backbone in the solid state. [N[(C3F7)C(Dipp)N]2]H reacts with copper(I) oxide in acetonitrile leading to [N[(C3F7)C(Dipp)N]2]CuNCCH3. This copper adduct serves as an excellent precursor to obtain thermally stable [N[(C3F7)C(Dipp)N]2]CuCNBut and [N[(C3F7)C(Dipp)N]2]CuCO. IR spectroscopic data of these copper(I) isocyanide (CN = 2176 cm(-1)) and copper(I) carbonyl (CO = 2109 cm(-1)) complexes indicate that the [N[(C3F7)C(Dipp)N]2]- ligand is a fairly weak donor. PMID- 15360224 TI - Simple generation of cationic aluminum alkyls and alkoxides based on the pendant arm tridentate schiff base. AB - The prepared in situ methyl(chloro)aluminum complex (2) from Me2AlCl and the pendant arm tridentate Schiff base (H-SchNMe2) was used to generate the methylaluminum cationic species [(SchNMe2)AlMe]+ in further reaction with 1 equiv of AlCl3 or NaBPh4 as the chloride abstracting reagents. The exposure of the resulting methylaluminum cationic species to an excess of dry dioxygen at 0 degrees C afforded the alkoxyaluminum cationic species, [(SchNMe2)AlOMe]+ or [(SchNMe2)AlOPh]+. The alkoxylaluminum cations proved to be a very efficient catalyst in the polymerization of epsilon-caprolactone. PMID- 15360225 TI - Formation of hexacoordinate complexes of PhCCSiF3 with 2,2'-bipyridine and with 1,10-phenanthroline through intermolecular silicon...nitrogen interactions. AB - Bidentate intermolecular Si...N interactions were utilized to form new hypervalent complexes of trifluoro-phenylethynyl-silane with 2,2'-bipyridine and with 1,10-phenanthroline. X-ray structures obtained for these complexes display a somewhat distorted octahedral geometry about the silicon atom. Binding constants ranging from 170 to 1600 M(-1) at 25 degrees C in CDCl3 were measured for the formation of these complexes, suggesting that such hypervalent complexes of silicon could be used as new motifs in supramolecular chemistry. PMID- 15360226 TI - Trapping an elusive intermediate in manganese-oxo cubane chemistry. AB - A new member of the Mn-oxo cubane core complex family [Mn2III,2IV4O4L6] (1), where L = (p-MeOPh)2PO2-, has been synthesized and characterized. Compound 1 possesses structurally inequivalent MnIII and MnIV with clear valence electron localization in the crystal phase, quite unlike the structurally equivalent sites, tetrahedral core symmetry, and delocalized valence of its analogue where L = Ph2PO2-. Compound 1 exhibits appreciable positive shifts (0.1-0.3 V) of both the oxidation and reduction electrochemical potentials, attributable to the remote electron donating p-MeO groups. Reduction of 1 by methanol yields a novel core complex [MnIII4O2(OMe)2(MeOH)[(p-MeOPh)2PO2]6] (2). Each MnIII of 2 is tetragonally distorted due to the Jahn-Teller effect with elongated Mn-O bonds that are directed at the two micro3-MeO bridges and neither of the two micro3 oxos. These electronically driven distortions provide a structural rationale for the greater basicity of the former sites and explain why 2 of the 4 corner oxos are preferentially reduced to water molecules. PMID- 15360227 TI - Accommodation of the irregular coordination geometry of lead(II) by a square planar N2S2 ligand and its preference for zinc(II). AB - The N2S2 ligand, bis-mercaptoethanediazacyclooctane, coordinates to Pb(II) largely through sulfur donors, enlisting a second unit to fulfill an irregular, hemispherical N2S3 coordination environment in which a void suggests the location of a stereochemically active lone pair on Pb(II). That the highly exposed lead is vulnerable to metal ion displacement is demonstrated on reaction with zinc which results in a regular square pyramidal coordination about zinc within a [N2S2Zn]2 dimer. Analysis of the two dimeric structures finds different connectivities of the monomeric subunits account for the stability of the zinc structure over that of the lead. PMID- 15360228 TI - A new redox-active coordination polymer with cobalticinium dicarboxylate. AB - A new two-dimensional coordination polymer with cobalticinium 1,1'-dicarboxylate (ccdc) incorporated in the framework has been prepared, the ccdc functioning as unique monoanionic dicarboxylate ligands. The compound shows a high redox activity based on the ccdc units. PMID- 15360229 TI - Nucleophilic aromatic substitution on aryl-amido ligands promoted by oxidizing osmium(IV) centers. AB - Addition of amine nucleophiles to acetonitrile solutions of the OsIV anilido complex TpOs(NHPh)Cl2 (1) [Tp = hydrotris(1-pyrazolyl)borate] gives products with derivatized anilido ligands, i.e., TpOs[NH-p-C6H4(N(CH2)5)]Cl2 (2) from piperidine and TpOs[NH-p-C6H4N(CH2)4]Cl2 (3) from pyrrolidine. These materials are formed in approximately 30% yield under anaerobic conditions, together with approximately 60% yields of the OsIII aniline complex TpOs(NH2Ph)Cl2 (5). Formation of the para-substituted materials 2 or 3 from 1 involves oxidative removal of two hydrogen atoms (two H+ and two e-). The oxidation can be accomplished by 1, forming 5, or by O2. Related reactions have been observed with other amines and with the 2-naphthylamido derivative, which gives an ortho substituted product. Kinetic studies indicate an addition-elimination mechanism involving initial attack of the amine nucleophile on the anilido ligand. These are unusual examples of nucleophilic aromatic substitution of hydrogen. Ab initio calculations on 1 show that the LUMO has significant density at the ortho and para positions of the anilido ligand, resembling the LUMO of nitrobenzene. By analogy with nucleophilic aromatic substitution, 2 is quantitatively formed from piperidine and the p-chloroanilide TpOs(NH-p-C6H4Cl)Cl2 (7). Binding the anilide ligands to an oxidizing OsIV center thus causes a remarkable umpolung or inversion of chemical character from a typically electron-rich anilido to an electron-deficient aromatic functionality. This occurs because of the coupling of redox changes at the TpOsIV center with bond formation at the coordinated ligand. PMID- 15360230 TI - EDTA and mixed-ligand complexes of tetravalent and trivalent plutonium. AB - EDTA forms stable complexes with plutonium that are integral to nuclear material processing, radionuclide decontamination, and the potentially enhanced transport of environmental contamination. To characterize the aqueous Pu(4+/3+)EDTA species formed under the wide range of conditions of these processes, potentiometry, spectrophotometry, and cyclic voltammetry were used to measure solution equilibria. The results reveal new EDTA and mixed-ligand complexes and provide more accurate stability constants for previously identified species. In acidic solution (pH < 4) and at 1:1 ligand to metal ratio, PuY (where Y4- is the tetra anion of EDTA) is the predominant species, with an overall formation constant of log beta110 = 26.44. At higher pH, the hydrolysis species, PuY(OH)- and PuY(OH)(2)2-, form with the corresponding overall stability constants log beta(11 - 1) = 21.95 and log beta(11 - 2) = 15.29. The redox potential of the complex PuY at pH = 2.3 was determined to be E(1/2) = 342 mV. The correlation between redox potential, pH, and the protonation state of PuY- was derived to estimate the redox potential of the Pu(4+/3+)Y complex as a function of pH. Under conditions of neutral pH and excess EDTA relative to Pu4+, PuY(2)4- forms with an overall formation constant of log beta120 = 35.39. In the presence of ancillary ligands, mixed-ligand complexes form, as exemplified by the citrate and carbonate complexes PuY(citrate)3- (log beta1101 = 33.45) and PuY(carbonate)2- (log beta1101 = 35.51). Cyclic voltammetry shows irreversible electrochemical behavior for these coordinatively saturated Pu4+ complexes: The reduction wave is shifted approximately -400 mV from the reduction wave of the complex PuY, while the oxidation wave is invariant. PMID- 15360231 TI - Structure, reactivity and aromaticity of acenes and their BN analogues: a density functional and electrostatic investigation. AB - Density functional calculations have been carried out on a series of linearly annelated acenes and their BN analogues. Even though borazine shows aromatic and reactivity behavior parallel with that of benzene, its condensed derivatives show patterns different from those of their hydrocarbon analogues. Nucleus independent chemical shift (NICS) values in acenes suggest that the aromaticity of the inner rings is more than that of benzene, whereas in BN-acenes there is no substantial change in the aromaticity of the individual rings. Molecular electrostatic potential (MESP) is employed to obtain further insights into the bonding and reactivity trends for these systems. The MESP topography patterns of acenes and BN-acenes are substantially different, with BN-acenes showing more localized pi electron features compared to those of acenes. The MESP values at the critical points (CPs) indicate overall lowering of aromaticity in these annelated systems. However, this change is gradual among the BN-acenes. PMID- 15360232 TI - Sulfur bridging interactions of cis-planar NiII-S2N2 coordination units with nickel(II), copper(I,II), zinc(II), and mercury(II): a library of bridging modes, including NiII(micro2-SR)2MI,II rhombs. AB - Sulfur bridging interactions between three cis-planar NiII-S2N2 complexes and NiII, CuI,II, ZnII, and HgII reactants were investigated by synthesis and X-ray crystal structures of some 24 complexes. This work was stimulated by recent crystallographic structures of the A-cluster of carbon monoxide dehydrogenase/acetylcoenzyme A synthase. This bridged biological assembly has the minimal formulation [Fe4S4]-(micro2-SCys)-[M((micro2-SCys)2Gly)Ni] with M = NiII, CuI, and ZnII at sites distal and proximal, respectively, to the iron-sulfur cluster. Bridges supported by representations of the distal nickel site were sought by reactions of the complexes [NiII(LH-S2N2)]2- and [NiII(LR-S2N2)], with 5-5-5 chelate ring patterns. Reaction products implicate the bridges Ni-(micro2 S)1,2-M in a variety of molecular structures, some with previously unknown connectivities of bridge atoms. The most frequently encountered bridge units are the nonplanar rhombs Ni(2-S)2M involving both sulfur atoms of a given complex. Those with M = NiII are biologically relevant inasmuch as the catalytic metal at the proximal site is nickel. The complex [Ni(L-655)]2-, containing the 6-5-5 ring pattern and coordination sphere of the distal nickel site, was prepared and structurally characterized. It was shown to sustain Ni2(micro2-S)2 rhombic interactions in the form of trinuclear [[Ni(L-655)]2Ni]2- and [[Ni(L 655)]Ni(R2PCH2CH2PR2)] (R = Et, Ph) in which the second NiII simulates the proximal site. Bridging interactions of NiII-S2N2 complexes are summarized, and geometrical features of Ni2(2-S)2 rhombs in these complexes, as dependent on ring patterns, are considered (LH-S2N2 = N,N'-ethylenebis(2-mercaptoisobutyramide)(4 ); LR-S2N2 = trans-rac-N,N'-bis(2-mercapto-2-methylprop-1-yl)-1,2 cyclohexanediamine(2-); L-655 = N-(2-mercaptopropyl)-N'-(2' mercaptoethyl)glycinamide(4-)). PMID- 15360233 TI - Functionalized metal oxide clusters: synthesis, characterization, crystal structures, and magnetic properties of a novel series of fully reduced heteropolyoxovanadium cationic clusters decorated with organic ligands- [MVIV6O6[(OCH2CH2)2N(CH2CH2OH)]6]X (M = Li, X = Cl x LiCl; M = Na, X = Cl x H2O; M = Mg, X = 2Br x H2O; M = Mn, Fe, X = 2Cl; M = Co, Ni, X = 2Cl x H2O). AB - A novel series of fully reduced heteropolyoxovanadium(IV) compounds, [MVIV6O6[(OCH2CH2)2N(CH2CH2OH)]6]X (1, M = Li, X = Cl x LiCl; 2, M = Na, X = Cl x H2O; 3, M = Mg, X = 2Br x H2O; 4, M = Mn, X = 2Cl; 5, M = Fe, X = 2Cl; 6, M = Co, X = 2Cl x H2O; 7, M = Ni, X = 2Cl x H2O), have been synthesized and characterized by FT-IR and UV-vis spectroscopies, thermogravimetric analysis, elemental analysis, manganometric titration, temperature-dependent magnetic susceptibility measurements, bond valence sum calculations, X-ray powder diffraction, and single crystal X-ray diffraction analyses. The structures of the crystals are comprised of discrete units of fully reduced cluster cations, [MVIV6O6[(OCH2CH2)2N(CH2CH2OH)]6]n+, counterions (chloride or bromide), and water of crystallization (in the case of 2, 3, 6, 7). In each case the cluster ion is composed of a fully reduced cyclic [MV6N6O18] (M = Li, Na, Mg, Mn, Fe, Co, Ni) framework decorated with six triethanolamine ligands. Two arms of each triethanolamine ligand are coordinated to the metallacycle, and the third arm projects outward from the hexagonal ring. The [MV6N6O18] core adopts the Anderson type structure. The cyclic core is comprised of a ring of six edge-sharing [VO5N] octahedra linked to a central [MO6] unit. The hexametalate ring contains six d1 ions [VIV] and shows remarkable flexibility to encapsulate a variety of metal centers Mn+ (Mn+ = Li+, Na+, Mg2+, Mn2+, Fe2+, Co2+, Ni2+) with different (dn) spins. The compounds show good thermal stability and exhibit interesting magnetic properties that make these magnetic clusters promising building blocks for constructing supramolecular structures and extended structure magnetic solids. Crystal data for 1; C36H78Cl2N6Li2O24V6, trigonal space group R, a = 13.7185(3) angstroms, c = 24.8899(8) angstroms, Z = 3. Crystal data for 2: C36H80ClN6NaO25V6, triclinic space group P, a = 11.1817(5) angstroms, b = 12.1612(5) angstroms, c = 21.5979(10) angstroms, alpha = 75.8210(10), beta = 78.8270(10), gamma = 71.1400(10), Z = 2. Crystal data for 4: C36H78Cl2N6MnO24V6, monoclinic, space group P2(1), a =11.2208(5) angstroms, b = 21.5041(9) angstroms, c = 11.8126(5), beta = 111.2680, Z= 2. Crystal data for 5: C36H78Cl2N6FeO24V6, monoclinic, space group P2(1), a = 11.3057(7) angstroms, b = 21.4372(13) angstroms, c = 11.8167(7) angstroms, beta = 111.4170, Z = 2. PMID- 15360234 TI - Au2(SeO3)2(SeO4): synthesis and characterization of a new noncentrosymmetric selenite-selenate. AB - The reaction of elemental gold and selenic acid in Teflon-lined steel autoclaves leads to orange-yellow single crystals of Au2(SeO3)2(SeO4) (orthorhombic, Z = 4, Cmc2(1) (No. 36), a = 1689.1(3) pm, b = 630.13(8) pm, c = 832.7(1) pm, V = 886.2(2) angstroms3, Rall = 0.0452). In the crystal structure, Au3+ is surrounded by four oxygen atoms of just as many monodentate SeO3(2-) ions in a square planar manner. The linkage of the polyhedra leads to double chains in the [001] direction which are connected to puckered layers by SeO4(2-) groups. The noncentrosymmetric space group could be proved by the observation of an SHG effect upon irridation at 1064 nm that shows an efficiency of about 43% compared to a KDP reference. Upon heating, Au2(SeO3)2(SeO4) decomposes at about 370 degrees C in one step yielding elemental gold. The presence of selenite and selenate groups in the compounds is also obvious from the IR and Raman spectra which show the characteristic bands of both species. Furthermore, solid-state NMR spectra reveal the different surroundings of the selenium atoms in the compound. PMID- 15360235 TI - Spontaneous self-assembly of an unsymmetric trinuclear triangular copper(II) pyrazolate complex, [Cu3(micro3-OH)(micro-pz)3(MeCOO)2(Hpz)] (Hpz = pyrazole). synthesis, experimental and theoretical characterization, reactivity, and catalytic activity. AB - The almost quantitative formation of the triangular trinuclear copper derivative [Cu3(3-OH)(-pz)3(MeCOO)2(Hpz)] (1) (Hpz = pyrazole), has been simply achieved by adding Hpz to an ethanol solution of Cu(MeCOO)2 x H2O. An X-ray molecular structure determination shows that 1 is completely unsymmetric and that trinuclear units result assembled in an extended bidimensional network formed through acetate bridges and hydrogen bonds. EPR and magnetic measurements are consistent with the presence of a single unpaired electron. Theoretical density functional calculations carried out for S = 1/2 provide a thorough description of the electronic structure of 1, allowing a detailed assignment of its UV-vis absorption spectrum. Compound 1 reacts with MeONa, yielding [Cu3(micro3-OH)(micro pz)3(MeCOO)(MeO)(Hpz)] (2) and [Cu3(micro3-OH)(micro-pz)3(MeO)2(Hpz)] (3) through the substitution of one and two acetate ions, respectively, with MeO- ion(s). The spontaneous self-assembly of the triangular trinuclear Cu3 moiety seems to occur only with pyrazole as can be inferred by the results obtained in the reactions of copper(II) acetate with some substituted pyrazoles leading to the formation of mononuclear [Cu(MeCOO)2(L)2] (4-8) and dinuclear [Cu(MeCOO)2(L)]2 (9-11) (L = substituted pyrazole) compounds. Also the presence of acetate ions seems to play a leading role in determining the formation of the trinuclear triangular arrangement, as indicated by the formation of a mononuclear derivative, [Cu(CF3COO)2(Hpz)]2 (compound 12), in the reaction of copper(II) trifluoroacetate with pyrazole. Compounds 1-3, as well as some other mono- and dinuclear copper(II)-substituted pyrazole complexes, have been tested as catalyst precursors in cyclopropanation reaction, observing the formation of products in a syn:anti ratio opposite that normally reported. PMID- 15360236 TI - Ultrasonic-assisted synthesis of monodisperse single-crystalline silver nanoplates and gold nanorings. AB - A simple sonochemical route was developed for the crystal growth of uniform silver nanoplates and ringlike gold nanocrystals in a N,N-dimethylformamide solution. The platelike structures were generated from the selective growth on different crystal planes in the presence of poly(vinylpyrrolidone) and the ultrasonic-assisted Ostwald ripening processes. The silver nanoplates in solution served as the templates for the synthesis of ringlike gold crystals via a displacement reaction. Both the silver nanoplates and gold nanorings were highly oriented single crystals with (111) planes as the basal planes. PMID- 15360237 TI - Formation of complex three- and one-dimensional interpenetrating networks within carbodiimide chemistry: NCN2(-)-coordinated rare-earth-metal tetrahedra and condensed alkali-metal iodide octahedra in two novel lithium europium carbodiimide iodides, LiEu2(NCN)I3 and LiEu4(NCN)3I3. AB - Orange-red transparent single crystals of LiEu2(NCN)I3 and LiEu4(NCN)3I3 were synthesized from fluxes of europium iodide, sodium cyanide, sodium azide, and lithium iodide at elevated temperatures and structurally characterized by X-ray diffraction. While LiEu2(NCN)I3 crystallizes in the cubic system (Fdm, a = 15.1427(17) angstroms, V = 3472.2(7) angstroms3, Z = 16, R1 = 0.0322), LiEu4(NCN)3I3 adopts the hexagonal system (P6(3)/mmc, a = 10.6575(11) angstroms, c = 6.8232(10) angstroms, V = 671.16(14) angstroms3, Z = 2, R1 = 0.0246). Both extended structures are composed of complex frameworks built from europium tetrahedra coordinated by carbodiimide (symmetrical NCN2-) units on the one side and condensed iodine octahedra around lithium cations on the other. Within LiEu2(NCN)I3, vertex-sharing of the LiI6 subunits together with isolated europium tetrahedra results in two three-dimensional networks interpenetrating each other. Within LiEu4(NCN)3I3, face-sharing of the europium tetrahedra results in bitetrahedral units which further connect via two opposing vertexes into one dimensional linkages. Likewise, the LiI6 octahedra share common faces to also yield one-dimensional linkages that fill the channels between the europium/carbodiimide substructure. The magnetic behavior of both compounds has been determined; the two phases follow Curie-Weiss laws with weak predominantly ferromagnetic exchange between the europium ions and atomic moments characterizing them as essentially divalent. PMID- 15360238 TI - Solvation of uranyl(II) and europium(III) cations and their chloro complexes in a room-temperature ionic liquid. A theoretical study of the effect of solvent "humidity". AB - We report a molecular dynamics study of the solvation of the UO2(2+) and Eu3+ cations and their chloro complexes in the [BMI][PF6][H2O] "humid" room temperature ionic liquid (IL) composed of 1-butyl-3-methylimidazolium+ and PF6- ions and H2O in a 1:1:1 ratio. When compared to the results obtained in dry [BMI][PF6], the present results reveal the importance of water. The "naked" cations form UO2(H2O)5(2+) and Eu(H2O)9(3+) complexes, embedded in a shell of 7 and 8 PF6- anions, respectively. All studied UO2Cln(2-n) and EuCln(3-n) chloro complexes remain stable during the dynamics and coordinate additional H2O molecules in their first shell. UO2Cl4(2-) and EuCl6(3-) are surrounded by an "unsaturated" water shell, followed by a shell of BMI+ cations. According to an energy component analysis, the UO2Cl4(2-) and EuCl6(3-) species, intrinsically unstable toward dissociation, are more stable than their less halogenated analogues in the IL solution, due to the solvation forces. The different chloro species also interact better with the humid than with the dry IL, which hints at the importance of solvent humidity to improve their solubility. Humidity markedly modifies the local ion environment, with major consequences as far as their spectroscopic properties are concerned. We finally compare the aqueous interface of [BMI][PF6] and [OMI][PF6] ionic liquids, demonstrating the importance of imidazolium substituents (N-butyl versus N-octyl) to the nature of the interface and miscibility with water. PMID- 15360239 TI - Transport of alkali halides through a liquid organic membrane containing a ditopic salt-binding receptor. AB - A ditopic receptor is shown to have an impressive ability to recognize and extract the ion pairs of various alkali halides into organic solution. X-ray diffraction analysis indicates that the salts are bound in the solid state as contact ion pairs. Transport experiments, using a supported liquid membrane and high salt concentration in the source phase, show that the ditopic receptor can transport alkali halide salts up to 10-fold faster than a monotopic cation or anion receptor and 2-fold faster than a binary mixture of cation and anion receptors. All transport systems exhibit the same qualitative order of ion selectivity; that is, for a constant anion, the cation selectivity order is K+ > Na+ > Li+, and for a constant cation, the anion transport selectivity order is I- > Br- > Cl-. The data suggest that with a ditopic receptor, the polarity of the receptor-salt complex can be lowered if the salt is bound as an associated ion pair, which leads to a faster diffusion through the membrane and a higher maximal flux. PMID- 15360240 TI - New route to the mixed valence semiquinone-catecholate based mononuclear FeIII and catecholate based dinuclear MnIII complexes: first experimental evidence of valence tautomerism in an iron complex. AB - The semiquinone-catecholate based mixed valence complex, [FeIII(bispicen)(Cl4Cat)(Cl4SQ)] x DMF (1), and catecholate based (H2bispictn)[Mn2III(Cl4Cat)4(DMF)2] (2) (bispicen = N,N'-bis(2-pyridylmethyl)-1,2 ethanediamine, bispictn = N,N'-bis(2-pyridylmethyl)-1,3-propanediamine, Cl4Cat = tetrachlorocatecholate dianion, and Cl4SQ = tetrachlorosemiquinone radical anion) were synthesized directly utilizing a facile route. Both the complexes have been characterized by single crystal X-ray diffraction study. The electronic structures have been elucidated by UV-vis-NIR absorption spectroscopy, cyclic voltammetry, EPR, and magnetic properties. The structural as well as spectroscopic features support the mixed valence tetrachlorosemiquinone tetrachlorocatecholate charge distribution in 1. The ligand based mixed valence state was further confirmed by the presence of an intervalence charge transfer (IVCT) band in the 1900 nm region both in solution and in the solid. The intramolecular electron transfer, a phenomenon known as valence tautomerism (VT), has been followed by electronic absorption spectroscopy. For 1, the isomeric form [FeIII(bispicen)(Cl4Cat)(Cl4SQ)] is favored at low temperature, while at an elevated temperature, the [FeII(bispicen)(Cl4SQ)2] redox isomer dominates. Infrared as well as UV-vis-NIR spectral characterization for 2 suggest that the MnIII(Cat)2- moiety is admixed with its mixed valence semiquinone-catecholate isomer MnII(SQ)(Cat)-, and the electronic absorption spectrum is dominated by the mixed charged species. The origin of the intervalence charge transfer band in the 1900 nm range is associated with the mixed valence form, MnII(Cl4Cat)(Cl4SQ)-. The observation of VT in complex 1 is the first example where a mixed valence semiquinone-catecholate iron(III) complex undergoes intramolecular electron transfer similar to manganese and cobalt complexes. PMID- 15360241 TI - Mixed-valence MnIIIMnIV clusters [Mn7O8(O2SePh)8(O2CMe)(H2O)] and [Mn7O8(O2SePh)9(H2O)]: single-chain magnets exhibiting quantum tunneling of magnetization. AB - The syntheses, structures, and magnetic properties of two new Mn7 complexes containing phenylseleninate ligands are reported. [Mn7O8(O2SePh)8(O2CMe)(H2O)] (1) and [Mn7O8(O2SePh)9(H2O)] (2) were both prepared by the reaction of 18 equiv of benzeneseleninic acid (PhSeO2H) with [Mn12O12(O2CMe)16(H2O)4] in MeCN. Complex 1 x 6MeCN crystallizes in the triclinic space group P, and complex 2 x 2CH2Cl2 crystallizes in the monoclinic space group P2(1)/m. Both compounds possess an unprecedented [Mn7O8]9+ core comprising a central [MnIII3(micro3-O)4]+ unit attached to [MnIV2(micro-O)2]4+ and [MnIV2(micro-O)(micro3-O)]4+ units on either side. In each cluster, the PhSeO2- groups function as bridging ligands between adjacent Mn centers. The structure reveals strong Se.O intermolecular contacts between Mn7 units to give a one-dimensional chain structure, with weak interchain interactions. Solid-state DC magnetic susceptibility measurements of complexes 1 and 2 reveal that they have very similar properties, and detailed studies on 1 by AC susceptibility measurements confirm an S = 2 ground-state spin value. In addition, out-of-phase AC signals are observed, suggesting slow magnetization relaxation. Magnetization versus DC field sweeps down to 0.04 K reveals hysteresis loops, but the temperature dependence of the coercivity is not what is expected of a single-molecule magnet. Instead, the behavior is due to single chain magnetism, albeit with weak antiferromagnetic interactions between the chains, with the barrier to relaxation arising from a combination of molecular anisotropy and ferromagnetic intermolecular exchange interactions mediated by the Se...O contacts. An Arrhenius plot was constructed from the magnetization versus time decay data. The thermally activated region at > 0.5 K gave an effective relaxation barrier (Ueff) of 14.2 K. Below approximately 0.1 K, the relaxation is independent of temperature, which is characteristic of magnetization quantum tunneling through the anisotropy barrier. These Mn7 compounds are thus the first single-chain magnets to comprise polynuclear metal clusters and also the first for which the temperature-independent relaxation characteristic of tunneling has been identified. The work also emphasizes that out-of-phase AC signals for ostensibly molecular compounds are not sufficient proof by themselves of a single molecule magnet. PMID- 15360242 TI - Intra and intermolecular magnetic interactions in a series of dinuclear Cu(II)/hxta complexes [H5hxta = N,N'-(2-hydroxy-1,3-xylylene)-bis-(N carboxymethylglycine)]: correlation of magnetic properties with geometry. AB - Nine dinuclear copper(II) complexes with hxta5- ligands [H5hxta = N,N'-(2-hydroxy 1,3-xylylene)-bis-(N-carboxymethylglycine)]: [Cu2(MeO-hxtaH)(H2O)2] x 4H2O (1), [Na(micro-H2O)2(H2O)6][Cu2(Cl-hxta)(H2O)3]2 x 6H2O (2), [Cu(H2O)6][Cu2(Me hxta)(H2O)2](NO3) x 2H2O (3), [Cu2(R-hxtaH)(H2O)3] x 3H2O [R = Cl (4), CH3 (5), and MeO (6)], [Cu2(MeO-hxtaH2)(micro-X)(CH3OH)] x 3CH3OH [X = Cl (7), Br (8)] and K5Na(micro-H2O)10[Cu2(micro-CO3)(Me-hxta)]2 x 4H2O (9), have been synthesized and structurally characterized. In complexes 4-7, the dinuclear units are linked via novel pairwise supramolecular interactions involving the ligand carboxylate groups. The intra- and intermolecular magnetic interactions have been quantified, and the coupling constants have been related to the structural geometries. PMID- 15360243 TI - Hybrid DFT study of the mechanism of quercetin 2,3-dioxygenase. AB - The mechanism of the copper-containing enzyme quercetin 2,3-dioxygenase has been studied using hybrid density functional theory. This enzyme cleaves the O heterocycle of a flavonol using dioxygen and releases carbon monoxide. Two different pathways for the dioxygen attack on the copper complex have been investigated, and the one where the first attack is on copper is found to be the energetically preferred one. By using this pathway the problem of having to go through a spin-orbit-induced spin crossing is also avoided. The adduct has three unpaired spins and is ideally suited for forming a dioxygen bridging structure, which occurs in the next step. Rather than cleaving the O-O bond in the next step, another C-O bond between dioxygen and the substrate is first formed. Finally, the O-O bond is cleaved, and CO is released in one concerted transition state with a very low barrier. The results are in good agreement with experimental findings. The mechanism is compared to the ones for other similar enzymes studied recently by similar methods. PMID- 15360244 TI - New aluminate with a tetrahedral structure closely related to the c84 fullerene. AB - A new aluminate Sr33Bi(24+delta)Al48O(141+3delta/2), having an F3m cubic structure (a = 25.090 angstroms, Z = 4) and forming a close packed face centered cubic array of "Al84" fullerene geometry, has been discovered. This original structure consists of corner-sharing AlO4 tetrahedra forming "Al84O106" cubic units whose assemblage delimits five types of cages, three of them being empty, one being occupied by strontium, and the fifth one forming the huge spheric fullerene-type cavity. In the latter, strontium, oxygen, and bismuth ions form concentric spheres, with an onion-skin-like configuration. The latter ions are disposed into a compact "Bi16O(52-n[]n)" anion whose the exceptional geometry is characterized by a strong stereoactivity of the 6s2 lone pair of Bi3+. PMID- 15360245 TI - Theoretical studies of ground and excited electronic States in a series of rhenium(i) bipyridine complexes containing diarylethynyl-based structure. AB - The photophysical properties, which vary as X is varied, of Re(I)-halide complexes (X2-bpy)ReICl(CO)3 (where X=ph, DAE, DNE, and DPE; ph = phenyl (1); DAE = di(amineoethynylbenzene) (2); DPE = di(phenylethynylbenzene) (3); DNE = di(nitroethynylbenzene) (4); bpy=2,2'bipyridine), are investigated using density functional theory (DFT). The electronic properties of the neutral molecules, in addition to the positive and negative ions, are studied using B3LYP functional. Excited singlet and triplet states are examined using time-dependent density functional theory (TDDFT). The low-lying excited-state geometries are optimized at the ab initio configuration interaction singlets level. As shown, the diarylethynyl-based structure is an integral component of the bpy pi-conjugated network, which results in a good planar structure. The occupied orbitals involved in the transitions have a significant mixture of metal Re and group Cl, and the lowest unoccupied orbital is a pi orbital, which extends from ligand bpy to diarylethynyl-based substituents. The luminescence for each complex originates from the lowest triplet excited states and is assigned to the mixing of MLCT and LLCT characters. Significant insights on the effects of these diarylethynyl conjugated structure and ending substituents (NH2, ph, and NO2) on absorption and emission spectra are observed by analysis of the results of the TDDFT method. The diarylethynyl-based pi-conjugated network makes both the absorption and emission spectra red-shifted compared with simple complex (bpy)ReICl(CO)3. Furthermore, an electron-releasing group (NH2) makes absorption and emission spectra blue-shift and an electron-withdrawing group (NO2) makes them red-shift. PMID- 15360246 TI - Complex reduction chemistry of (abpy)PtCl2, abpy = 2,2'-azobispyridine: formation of cyclic [(micro,eta2:eta1-abpy)PtCl]2(2+) with a new coordination mode for abpy and a near-infrared ligand-to-ligand intervalence charge transfer absorption of the one-electron reduced state. AB - The structurally characterized (abpy)PtCl2, abpy = 2,2'-azobispyridine, reveals a strong metal/ligand pi interaction as supported by DFT calculations. Unexpectedly, its chemical or electrochemical reduction occurs irreversibly to yield EPR-detectable [(micro,eta2:eta2-abpy)[PtCl2]2]*- and, as the main product of chloride dissociation, the structurally identified and DFT-calculated dinuclear [(micro,eta2:eta1-abpy)PtCl]2(2+) with a novel coordination mode for abpy and isolated as tetrachlorozincate. Stepwise reversible one-electron reduction of that dimer, separated by 0.24 V, exhibits an intense near-infrared band for the monocationic intermediate [(abpy(-I))(abpy)Pt2Cl2]*+ at 1220 nm (epsilon = 3370 M(-1) cm(-1)) which is attributed to a ligand-to-ligand intervalence charge-transfer transition. PMID- 15360247 TI - Stabilized arsenic(i) iodide: a ready source of arsenic iodide fragments and a useful reagent for the generation of clusters. AB - The new stable low oxidation state arsenic(I) iodide reagent [(dppe)As][I] (dppe = 1,2-bis(diphenylphosphino)ethane) exhibits chemistry that is considerably different from its AsIII analogues. While [(dppe)As][I] is not crystalline, the crystal structure of the derivative salt [(dppe)As][(dppe)As2I7] is reported and is compared to that of [(dppe)As]2[SnCl6] x 2CH2Cl2. The air oxidation of [(dppe)As][I] produces crystals of the salt [Ph2P(O)CH2CH2P(OH)Ph2]2[As6I8] x 2CH2Cl2 and suggests that, in contrast to previous studies, the reaction of the univalent arsenic iodide salt with certain oxidants results in the oxidation of the dppe ligand and the release of "AsI-I" fragments that oligomerize to form AsI clusters. Such reactivity is confirmed by the reaction of 6[(dppe)As][I] with 12Me3NO and 2[PPh4][I] to produce [PPh4]2[As6I8] and 6dppeO2. The reactivity is rationalized using density functional theory calculations. PMID- 15360248 TI - Mono-, bi-, and trinuclear CuII-Cl containing products based on the tris(2 pyridylmethyl)amine chelate derived from copper(I) complex dechlorination reactions of chloroform. AB - The ligand TMPA (tris(2-pyridylmethyl)amine) and its copper complexes have played a prominent role in recent (bio)inorganic chemistry studies; the copper(I) complex [CuI(TMPA)(CH3CN)]+ possesses an extensive dioxygen reactivity, and it is also known to effect the reductive dechlorination of substrates such as dichloromethane and benzyl and allyl chlorides. In this report, we describe a set of new analogues of TMPA, ligand 6TMPAOH, binucleating Iso-DO, and trinucleating SYMM. Copper(I) complexes with these ligands and a previously described binucleating ligand DO react with chloroform, resulting in reductive dechlorination and production of [CuIIx(L)Clx]x+ (x = 1, 2, or 3). X-ray crystal structures of [CuII(6TMPAOH)Cl]PF6, [CuII2(Iso-DO)Cl2](PF6)2, [CuII2(DO)Cl2](PF6)2, and [Cu3(SYMM)Cl3](PF6)3 are presented, and the compounds are also characterized by UV-vis and EPR spectroscopies as well as cyclic voltammetry. The steric influence of a pyridyl 6-substituent (in the complexes with 6TMPAOH, Iso-DO, and SYMM) on the solid state and solution structures and redox potentials are compared and contrasted to those chlorocopper(II) complexes with a pyridyl 5'-substituent (in [CuII2(DO)Cl2](PF6)2 and in [CuII(TMPA)Cl]+). Some insights into the reductive dechlorination process have been obtained by using 2H NMR spectroscopy in following the reaction of [Cu2(Iso DO)(CH3CN)2](PF6)2 with CDCl3, in the presence or absence of a radical trap, 2,4 di-tert-butylphenol. PMID- 15360249 TI - Spectroscopic and structural characterizations of novel water-soluble tetraperoxo and diperoxo[polyaminocarboxylato bis(N-oxido)]tantalate(V) complexes. AB - New water-soluble homoleptic peroxo complexes and heteroleptic peroxo polyaminocarboxylato (PAC) complexes of tantalum(V) have been prepared. In the case of the peroxo-PAC complexes, the synthesis in the presence of excess H2O2 leads to the oxidation of the nitrogen atoms of the ligand into N-oxides. The compounds correspond to the general formula (gu)3[Ta(O2)2(LO2)] x xH2O (gu = guanidinium, L = edta or pdta) in which H4LO2 refers to the bis(N-oxide) derivative of the PAC ligand. The TaV complexes have been characterized on the basis of elemental and thermal analysis and by IR and 13C and 15N NMR spectroscopy. These last two spectroscopic methods have been used to suggest the coordination mode of the PAC ligand in the complexes. ESI mass spectrometry measurements have also been carried out for the peroxo-PAC compounds. The crystal structures of the homoleptic tetraperoxotantalate, (gu)3[Ta(O2)4] (1), and the heteroleptic complex, (gu)3[Ta(O2)2(edtaO2)] x 2.32H2O x 0.68H2O2 (2b), have been determined, showing, for both cases, an 8-fold-coordinated Ta atom surrounded either by four bidentate peroxides or by two peroxides and one tetradentate edtaO2 ligand. PMID- 15360250 TI - Synthesis, characterization, and structures of indium In(DTPA-BA2) and yttrium Y(DTPA-BA2)(CH3OH) complexes (BA = Benzylamine): models for 111In- and 90Y labeled DTPA-biomolecule conjugates. AB - To explore structural differences in In3+, Y3+, and Lu3+ chelates, we prepared M(DTPA-BA2) complexes (M = In, Y, and Lu; DTPA-BA2 = N,N' ' bis(benzylcarbamoylmethyl)diethylenetriamine-N,N',N' '-triacetic acid) by reacting the trisodium salt of DTPA-BA2 with 1 equiv of metal chloride or nitrate. All three complexes have been characterized by elemental analysis, HPLC, IR, ES-MS, and NMR (1H and 13C) methods. ES-MS spectral and elemental analysis data are consistent with the proposed formula for M(DTPA-BA2) (M = In, Y, and Lu) and have been confirmed by the X-ray crystal structures of both In(DTPA-BA2) x 2H2O and Y(DTPA-BA2)(CH3OH) complexes. By a reversed-phase HPLC method, it was found that In(DTPA-BA2) is more hydrophilic than M(DTPA-BA2) (M = Y and Lu), most likely due to the dissociation of the two carbonyl oxygen donors in solution. The X-ray crystal structure of In(DTPA-BA2) revealed a rare example of an eight coordinated In3+ complex with DTPA-BA2 bonding to the In3+ in a distorted square antiprism coordination geometry. Both benzylamine groups are in the trans position relative to the acetate-chelating arm that is attached to the central N atom. The Y3+ in Y(DTPA-BA2)(CH3OH) is nine-coordinated with an octadentate DTPA BA2 and a methanol oxygen. The coordination geometry is best described as a tricapped trigonal prism. One benzylamine group is trans and the other cis to the acetate-chelating arm that is attached to the central N atom. All three M(DTPA BA2) complexes (M = In, Y, and Lu) exist as at least three isomers in solution (approximately 10 mM), as shown by the presence of 6-8 overlapped 1H NMR signals from the methylene hydrogens of the benzylamine groups. The coordinated DTPA-BA2 remains rigid even at temperatures > 85 degrees C. The exchange rate between different isomers in M(DTPA-BA2) (M = In, Y, and Lu) is relatively slow at high concentrations (> 1.0 mM), but it is fast due to the partial dissociation and rapid interconversion of different isomers at lower concentrations ( approximately 10 mircroM). It is not surprising that M(DTPA-BA2) complexes (M = In, Y, and Lu) appear as a single peak in their respective HPLC chromatogram. PMID- 15360251 TI - Synthesis, structure, magnetism, and spectroscopic properties of heterobinuclear copper(II)-zinc(II) complexes and their copper(II)-copper(II) analogues in asymmetric ligand environments. AB - Heterobinuclear copper(II)-zinc(II) complexes and their homobinuclear dicopper(II) counterparts (1-4) of two asymmetric ligands (H2L1 and H2L2), based on 2-aminocyclopent-1-ene-1-dithiocarboxylate, are reported. The ligands are capable of providing both donor set and coordination number asymmetry in tandem. Metal centers in these complexes are connected by a micro-alkoxo and a bridging pyrazolate moiety, as confirmed by X-ray structure analyses of 1, 3, and 4. The Cu(1) site in the dicopper complex (1) is square planar and so are the copper sites in the Cu-Zn complexes 3 and 4. The pentacoordinated Zn sites in the latter complexes have distorted TBP geometry (tau = 0.74), while the corresponding Cu site in 1 has a highly distorted square pyramidal structure (tau = 0.54). The Cu...Zn separations in 3 and 4 are 3.3782 and 3.3403 angstroms, respectively, while the Cu...Cu distance in 1 is 3.3687 angstroms. The dicopper complexes are EPR silent at 77 K, in which the copper(II) centers are coupled by strong antiferromagnetic coupling (J = ca. -290 cm(-1)) as confirmed by variable temperature (4-300 K) magnetic measurements. These compounds (1 and 2) undergo two one-electron reductions and a single step two-electron oxidation at ca. 0.26, -1.40, and 1.0 V vs Ag/AgCl reference, respectively, as indicated by cyclic and differential pulse voltammetry done at subambient temperatures. EPR spectra of 3 and 4 display axial anisotropy at 77 K with the gperpendicular region being split into multiple lines due to N-superhyperfine coupling (AN = 15.3 x 10(-4) cm(-1)). The observed trend in the spin-Hamiltonian parameters, gparallel > gperpendicular > 2.04 and |Aperpendicular| << |Aparallel| approximately (120-150) x 10(-4) cm(-1), indicates a d(x2-y2)-based ground state with tetragonal site symmetry for the Cu(II) center in these molecules. PMID- 15360252 TI - Cyclohexene oxide/CO2 copolymerization catalyzed by chromium(III) salen complexes and N-methylimidazole: effects of varying salen ligand substituents and relative cocatalyst loading. AB - A detailed mechanistic study into the copolymerization of CO2 and cyclohexene oxide utilizing CrIII(salen)X complexes and N-methylimidazole, where H2salen = N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-ethylenediimine and other salen derivatives and X = Cl or N3, has been conducted. By studying salen ligands with various groups on the diimine backbone, we have observed that bulky groups oriented perpendicular to the salen plane reduce the activity of the catalyst significantly, while such groups oriented parallel to the salen plane do not retard copolymer formation. This is not surprising in that the mechanism for asymmetric ring opening of epoxides was found to occur in a bimetallic fashion, whereas these perpendicularly oriented groups along with the tert-butyl groups on the phenolate rings produce considerable steric requirements for the two metal centers to communicate and thus initiate the copolymerization process. It was also observed that altering the substituents on the phenolate rings of the salen ligand had a 2-fold effect, controlling both catalyst solubility as well as electron density around the metal center, producing significant effects on the rate of copolymer formation. This and other data discussed herein have led us to propose a more detailed mechanistic delineation, wherein the rate of copolymerization is dictated by two separate equilibria. The first equilibrium involves the initial second-order epoxide ring opening and is inhibited by excess amounts of cocatalyst. The second equilibrium involves the propagation step and is enhanced by excess cocatalyst. This gives the [cocatalyst] both a positive and negative effect on the overall rate of copolymerization. PMID- 15360253 TI - Synthesis and characterization of a series of rubidium alkoxides and rubidium titanium double alkoxides. AB - This report investigates the structural aspects of the products isolated from the reactions of a series of titanium alkoxides [[Ti(OR)4]n n = 2, OR = OCH2C(CH3)3 (ONep) (1); n = 1, OC6H3(CH3)2-2,6 (DMP) (2)] with rubidium alkoxides [[Rb(OR)]infinity where OR = (ONep) (3), (DMP) (4), and OC6H3(CH(CH3)2)2-2,6 (DIP) (5)]. The resultant double alkoxides were determined by single crystal X ray diffraction to be [Rb(mu-ONep)4(py)Ti(ONep)]2 (6), [Rb(mu DMP)Ti(DMP)4]infinity (7), and [Rb(mu-DMP)2(mu-ONep)2Ti(ONep)]infinity (8). Compound 1 is the previously reported dinculear species with trigonal bipyramidal Ti metal centers whereas compound 2 is a monomer with a tetrahedral Ti center. Suitable X-ray quality crystals of 3 were not isolated. Compounds 4 and 5 demonstrate extended polymeric networks with Rb coordination ranging from two to five utilizing terminal mu- and mu3-OR ligands and pi-interactions of neighboring OAr ligands. The double alkoxide 6 revealed a simple tetranuclear structure with mu-ONep acting as the bridge, terminal ONep ligands on the Ti, and one terminal py on the Rb. For 7 and 8, the pi-interaction facilitated the formation of extended polymeric systems. All complexes were further characterized by FT-IR and multinuclear NMR spectroscopy. PMID- 15360254 TI - Pyrazolylborate-zinc alkoxide complexes. 1. Basic properties, methylations, and heterocumulene insertions. AB - While a solution of TpPh,MeZn-OH in methanol contains only traces of TpPh,MeZn OMe, according to the equilibrium constant K = 5.8 x 10(-4), the reactions of TpPh,MeZn-OH with the electronegative alcohols trifluoroethanol and hexafluoro-2 propanol easily yield TpPh,MeZn-OCH2CF3 and TpPh,MeZn-OCH(CF3)2. The extremely hydrolytically sensitive TpPh,MeZn-OR complexes, with R = Me, Et, i-Pr, and CH2CH2F, as well as TpCum,MeZn-OR, with R = Me and i-Pr, are accessible from the Tp*Zn-hydride complexes and the corresponding alcohol. Alkylations with methyl iodide have revealed the high nucleophilicity of TpPh,MeZn-OMe by conversion to dimethyl ether and TpPh,MeZn-I. This conversion occurs rapidly not only with pure TpPh,MeZn-OMe but also with TpPh,MeZn-OH (as such or in the presence of methanol) and with TpPh,MeZn-OCOOMe. A relation of the Tp*Zn-alkoxides to the function of the zinc enzyme alcoholdehydrogenase exists in the reaction of TpPh,MeZn OCH(CH3)2 with aromatic aldehydes, which yields acetone and the corresponding benzyl oxides TpPh,MeZn-OH2Ar. The heterocumulenes carbon dioxide, carbon disulfide, isothiocyanates, and one isocyanate are inserted into the Zn-OR bonds, yielding one alkyl carbonate complex (TpPh,MeZn-OC(O)OMe), two xanthogenate complexes (TpPh,MeZn-SC(S)OR), three iminothiocarbonate complexes (TpPh,MeZn SC(NR')OR), and one alkyl carbamate complex (TpPh,MeZn-NR-COOMe). All insertion reactions can be described by a common mechanism involving a four-center intermediate in which the most basic heteroatom of the heterocumulene is attached to zinc. PMID- 15360255 TI - Pyrazolylborate-zinc alkoxide complexes. 2. Solvolytic chemistry. AB - The methoxides TpPh,MeZn-OMe and TpCum,MeZn-OMe were tested for their reactivity toward substrates that are hydrolytically cleaved with Tp*Zn-OH complexes. They do not induce the cleavage of nonactivated esters, phosphoesters, lactones, or lactams. They cleave the P-O-P linkage of tetraalkylpyrophosphates, but not the C O-C linkage of dialkyl pyrocarbonates. Transesterification of esters and phosphoesters occurs when they are activated as p-nitrophenolates. The most facile cleavage occurs for thiolate functions present in dithioesters, thiolactones, and trithiocarbonates. These findings indicate that, while the leaving group properties of the methoxide unit are essential, it is the strength of the resulting zinc-substrate bonds that decides upon the occurrence or nonoccurrence of the cleavage reactions. PMID- 15360256 TI - Pyrazolylborate-zinc alkoxide complexes. 3. Acid-base reactions. AB - The alkoxides TpPh,MeZn-OR (R = Me, Et, i-Pr) undergo acid-base reactions with all hydrogen compounds whose acidity is higher than that of the corresponding alcohol ROH. Thus, anion exchange occurs with the common acids acetic acid, acetohydroxamic acid, acetylacetone, phenol, and ethylmercaptan. Alkoxide exchange is observed using methanol, ethanol, and trifluoroethanol. With the NH acids cyanamide, trifluoroacetamide, and pyrazoles, the corresponding anions are attached to zinc, and likewise beta- and gamma-lactams, a thiazolidinedione, and the cyclic sulfimide saccharin are deprotonated. Of the CH acids acetonitrile forms the Tp*Zn-cyanomethanide. Acetone is deprotonated by the cyanomethanide complex and incorporated as the Tp*Zn-beta-ketoiminate. PMID- 15360257 TI - Synthesis of alpha-MoTe2 nanorods via annealing Te-seeded amorphous MoTe2 particles. AB - Semiconductor alpha-MoTe2 nanorods have been synthesized by annealing Te-seeded particles of an amorphous MoTe2 intermediate. This intermediate is prepared by a solution reaction between Mo(CO)6 and elemental Te in diphenylmethane. The as synthesized products were characterized by structural, compositional, and morphological techniques of X-ray diffraction, selected area electron diffraction, selected area energy dispersive spectroscopy, energy dispersive X ray analysis, X-ray photoelectron spectroscopy, transmission electron microscopy, and high-resolution transmission electron microscopy. The results of the annealing process are MoTe2 nanorods with diameters of 50-200 nm and lengths ranging from 0.1 to 3.0 microm. Here, the rodlike structure of MoTe2 is reported for the first time, and added to the list as one kind of new morphology of MoTe2 nanomaterials. A mechanism for the formation of the nanorods is proposed. The sandwich-layered structure of Te-Mo-Te and the similarity in the structure between hexagonal alpha-MoTe2 and hexagonal Te are responsible for the formation of nanorods of MoTe2. PMID- 15360258 TI - Synthesis, characterization, and dynamic studies of 12-vertex eta5-ruthenium(II) closo-phosphine complexes with monoanionic [10-L-nido-7-R-7,8-C2B9H9]- ligands. AB - Ruthenacarborane complexes of formula [3-H-3,3-(PPh3)2-8-L-closo-3,1,2 RuC2B9H10)] (L = SMe2 (2a), SEt2 (2b), S(CH2)4 (2c), SEtPh (2d)) and [1-Me-3-H 3,3-(PPh3)2-8-L-closo-3,1,2-RuC2B9H9)] (L = SMe2 (2e), SEt2 (2f)) were prepared by reaction of the respective monoanionic charge-compensated ligands [10-L-nido 7,8-C2B9H10]- and [7-Me-10-L-nido-7,8-C2B9H9]- with [RuCl2(PPh3)3]. Similary, complexes [3-H-3,3,8-(PPh3)3-closo-3,1,2-RuC2B9H10)] (4a) and [3-H-3,3-(PPh3)2-8 PPh2Me-closo-3,1,2-RuC2B9H10)] (4b) were prepared from the corresponding phosphonium ligands. The reaction is done in one pot by reacting the ligand with the Ru(II) complex in a 1.5:1 ratio. All compounds have been fully characterized by multinuclear NMR spectroscopy, and the molecular structures for 2a and 4a have been elucidated by single-crystal X-ray diffraction analysis. The Ru(II) atom in this complex is on the open face of the monoanionic charge-compensated ligand adopting a pseudooctahedral coordination. Formally, three positions are supplied by the C2B3 open face, two PPh3 groups occupy two other positions, and a hydride fulfills the remaining one. The hydride complexes were generated with no special reagent. They result from a dehalogenation in the presence of ethanol. PMID- 15360259 TI - Ferromagnetic anomaly associated with the antiferromagnetic transitions in (donor)[Ni(mnt)2]-type charge-transfer salts. AB - Three newly prepared [Ni(mnt)2] complexes, (HMTTF)[Ni(mnt)2], (ChSTF)[Ni(mnt)2], and (DBTTF)2[Ni(mnt)2], are reported (DBTTF = dibenzotetrathiafulvalene, ChSTF = 2,3-cyclohexylenedithio-1,4-dithia-5,8-diselanafulvalene, HMTTF = bis(trimethylene)-tetrathiafulvalene, and mnt = maleonitrile dithiolate). The former two compounds have usual DA-type (D = donor, A = acceptor) mixed stacks, whereas the DBTTF complex has DDDDAA-type 6-fold columns. These compounds are electrical insulators, but the HMTTF and ChSTF complexes exhibit chiT minima at 16 and 55 K, respectively, followed by chiT peaks at 8 and 16 K. Below these temperatures the ESR signal disappears, indicating antiferromagnetic transitions. The origin of the ferromagnetic interaction is explained either from the difference of the g values between the donor and the anion or from the intrinsic ferromagnetic interaction of the [Ni(mnt)2] anions. PMID- 15360260 TI - Room temperature phosphorescence from ruthenium(II) complexes bearing conjugated pyrenylethynylene subunits. AB - We describe the synthesis, electrochemistry, and photophysical properties of several Ru(II) complexes bearing different numbers of pyrenylethynylene substituents in either the 5- or 5,5'-positions of 2,2'-bipyridine, along with the appropriate Ru(II) model complexes bearing either bromo- or ethynyltoluene functionalities. In addition, we prepared and studied the photophysical behavior of the diimine ligands 5-pyrenylethynylene-2,2'-bipyridine and 5,5' dipyrenylethynylene-2,2'-bipyridine. Static and dynamic absorption and luminescence measurements reveal the nature of the lowest excited states in each molecule. All model Ru(II) complexes are photoluminescent at room temperature and exhibit excited-state behavior consistent with metal-to-ligand charge transfer (MLCT) characteristics. In the three Ru(II) molecules bearing multiple pyrenylethynylene substituents, there is clear evidence that the lowest excited state is triplet intraligand (3IL)-based, yielding long-lived room temperature phosphorescence in the red and near IR. This phosphorescence emanates from either 5-pyrenylethynylene-2,2'-bipyridine or 5,5'-dipyrenylethynylene-2,2'-bipyridine, depending upon the composition of the coordination compound. In the former case, the excited-state absorption difference spectra that were measured for the free ligand are easily superimposed with those obtained for the metal complexes coordinated to either one or two of these species. The latter instance is slightly complicated since coordination of the 5,5'-ligand to the Ru(II) center planarizes the diimine structure, leading to an extended conjugation on the long axis with a concomitant red shift of the singlet pi-pi absorption transitions and the observed room temperature phosphorescence. As a result, transient absorption measurements obtained using free 5,5'-dipyrenylethynylene-2,2'-bipyridine show a marked blue shift relative to its Ru(II) complex, and this extended pi conjugation effect was confirmed by coordinating this ligand to Zn(II) at room temperature. In essence, all three pyrenylethynylene-containing Ru(II) complexes are unique in this genre of chromophores since the lowest excited state is 3IL based at room temperature and at 77 K, and there is no compelling evidence of interacting or equilibrated excited states. PMID- 15360261 TI - Mechanistic elucidation of the substitution behavior of alkyl cobaloximes in water and methanol as solvents. AB - The ligand substitution behavior of trans-[Co(Hdmg)2(R)S] (R = CH3, PhCH2; Hdmg = dimethylglyoximate; S = H2O and/or MeOH) was studied for imidazole, pyrazole, 1,2,4-triazole, N-acetylimidazole, 5-chloro-1-methylimidazole, NO2-, Ph3P, Ph3As, and Ph3Sb as entering ligands. In all cases, except for Ph3As and Ph3Sb, the observed kinetics shows a linear dependence on the entering nucleophile concentration with no evidence for a back reaction. In the case of Ph3As and Ph3Sb as entering nucleophiles, kinetic evidence for a reverse solvolysis reaction is at hand. Activation parameters (DeltaH++, DeltaS++, and DeltaV++) were determined from the temperature and pressure dependence of all studied reactions and support the operation of a dissociatively activated substitution mechanism. The rate and activation parameters show that there is an increase in the dissociative character from a dissociative interchange to a limiting dissociative mechanism that depends on the nature of R and the entering nucleophile. The crystal structure of trans-[Co(en)2(Me)H2O]2+ was determined by X-ray analysis. The Co-O and Co-C bond lengths were found to be 2.153(6) and 1.995(10) angstroms, respectively. The kinetic and structural data are discussed in reference to a series of earlier studied systems for which data are reported in the literature. PMID- 15360262 TI - An unsymmetrical tripodal ligand with NOS2-donor set: coordination chemistry with nickel(II) and zinc(II). AB - The synthesis of the novel tripodal ligand [N(CH2CH2CH2OH)(CH2CH2SH)2] H3-4 is reported. The aliphatic tetradentate ligand is equipped with an unsymmetrical NOS2 donor set. It reacts with Ni(OAc)2 x 4H2O or Zn(BF4)2 x xH2O to give the complexes [Ni(H-4)]2 5 and [Zn(H-4)]4 6, respectively. The molecular structures of 5 and 6 have been determined by X-ray diffraction. In both cases multinuclear, mu-thiolato-bridged complexes, wherein the ligand coordinates with only three (NS2) of the four donor groups, had formed. The dinuclear complex 5 adopts a butterfly geometry and contains nickel(II) ions in a square-planar NS3 coordination environment. Cyclic voltammetry experiments indicate that the nickel centers in 5 are electron-rich but not overly sensitive toward oxidation. Complex 6 is tetranuclear and the four thiolato-bridged metal centers form a ring. It shows a distorted tetrahedral coordination geometry for the zinc(II) ions in an NS3 coordination sphere. In both complexes the hydroxyl functionalized ligand arm of the tripodal ligand remains uncoordinated. PMID- 15360263 TI - Isovalent and mixed-valent diruthenium complexes [(acac)2RuII ( bpytz)RuII(acac)2] and [(acac)2RuII(-bpytz)RuIII(acac)2](ClO4) (acac = acetylacetonate and bpytz = 3,6-bis(3,5-dimethylpyrazolyl)-1,2,4,5-tetrazine): synthesis, spectroelectrochemical, and epr investigation . AB - The title compounds involving the structurally characterized bridging ligand bpytz were characterized, showing very strong electrochemical stabilization of the mixed-valent RuIIRuIII state (Kc = 10(13.9)) but no detectable (epsilon < 20 M(-1) cm(-1)) intervalence charge-transfer band in the infrared region. In situ reduction of the neutral precursor produces a diruthenium(II) complex of the bpytz radical anion according to EPR spectroscopy, whereas oxidation of the mixed valent form leads to a diruthenium(III) species. PMID- 15360264 TI - Synthesis and H+, Cu2+, and Zn2+ coordination behavior of a bis(fluorophoric) bibrachial lariat aza-crown. AB - The synthesis, protonation behavior, and Cu2+ and Zn2+ coordination chemistry of the novel bibrachial aza lariat ether (naphthalen-1-ylmethyl)[2-(20-[2 [(naphthalen-1-ylmethyl)amino]ethyl]-3,6,9,17,20,23,29,30 octaazatricyclo[23.3.1.1*11,15*]triaconta-1(29),11(30),12,14,25,27-hexaen-6 yl)ethyl]amine (L) are discussed. The macrocycle, which has two aminoethyl naphthyl moieties symmetrically appended to a 2:2 azapyridinophane structure, displays, in the pH range 2-11, six protonation steps that correspond to the protonation of the secondary amino groups. Steady-state fluorescence measurements show emissions due to the monomer and to the excimer formed between the two naphthalene fragments of the macrocycle. The time-resolved fluorescence data, obtained by the time-correlated single photon counting technique, show that a significant percentage of excimer is preformed as ground-state dimers. The ligand L forms with the metal ions Cu2+ and Zn2+ mono- and dinuclear complexes in aqueous solution. The influence of metal coordination in the fluorescence emission of L is analyzed. The acid-base, coordination capabilities, and emissive behavior of L are compared with those presented by its synthetic precursor L1, which has a tripodal tris(2-aminoethyl)amine structure functionalized at one of its terminal amino groups with a naphthyl moiety. PMID- 15360265 TI - Poly(ortho esters)--from concept to reality. AB - The development of poly(ortho esters) dates back to the early 1970s, and during that time, four distinct families were developed. These polymers can be prepared by a transesterification reaction or by the addition of polyols to diketene acetals, and it is the latter method that has proven to be preferred one. The latest polymer, now under intense development, incorporates a latent acid segment in the polymer backbone that takes advantage of the acid-labile nature of the ortho ester linkages and allows control over erosion rates. By use of diols having selected chain flexibility, polymers that range from hard, brittle materials to materials that have a gel-like consistency at room temperature can be obtained. Drug release from solid materials will be illustrated with 5 fluorouacil and bovine serum albumin, and drug release from gel-like materials will be illustrated with mepivacaine, now in Phase II clinical trials as a delivery system to treat post-operative pain. A brief summary of preclinical toxicology studies is also presented. PMID- 15360266 TI - Amplification of inhibitory activity of catechin against disease-related enzymes by conjugation on poly(epsilon-lysine). AB - A new inhibitor against disease-related enzymes, collagenase, hyaluronidase, and xanthine oxidase, has been developed by the laccase-catalyzed conjugation of catechin on poly(epsilon-lysine). The resulting poly(epsilon-lysine)-catechin conjugate showed greatly improved inhibition effects on activity of these enzymes, whereas the catechin monomer showed very low inhibition activity. The kinetic analysis on the inhibition of collagenase exhibited that the conjugate was a mixed-type inhibitor. The amplified activities might offer high potential as a therapeutic agent for prevention of various enzyme-related diseases. PMID- 15360267 TI - Protein transfer through polyacrylamide hydrogel membranes polymerized in lyotropic phases. AB - A way to control the average pore size in cross-linked polyacrylamide-based membranes is by altering the ratio of cross-linker to acylamide monomer. Larger pore sizes are prepared with a minimum amount of cross-linker, resulting in membranes that are mechanically weak and have short lifetimes. The aim of this study was to prepare cross-linked polyacrylamide membranes with large pore sizes and with good mechanical integrity. The methodology was to carry out the polymerization in a template, formed from the self-aggregation of surfactant. Two surfactant templates were used, and their pore size was examined with proteins of different sizes. The surfactants chosen for this study were sodium dodecyl sulfate (SDS, ionic surfactant) and TERIC BL8 (nonionic surfactant), both of which have very different aggregation properties. The data showed that at 10% and greater of TERIC BL8, a very different and open gel structure is formed, in which the pore size was significantly increased. SDS seemed to have little effect on the pore size. The data suggests that the gel structures for both surfactants up to 4% (w/v) are similar and micellular, because SDS is known to favor a micelle structure. Above 4% (w/v), TERIC BL8 then goes through a change in its lyotropic phase, thus, producing membranes of a large pore size. In conclusion, the pore size and gel structure of polyacrylamide hydrogel membranes can be significantly increased using TERIC BL8 (nonionic) surfactant. This allows large-pore-size membranes with a high cross-link density and consequently high mechanical strength to be prepared for the separation of large biomolecules. PMID- 15360268 TI - Direct observation of poly(3-hydroxybutyrate) depolymerase adsorbed on polyester thin film by atomic force microscopy. AB - Poly[(R)-3-hydroxybutyrate] (PHB) depolymerases adsorbed on poly(L-lactide) (PLLA) thin film were directly observed by atomic force microscopy (AFM). A PLLA thin film of 100 nm thickness was prepared on a silicon wafer by spin-cast method. The PLLA thin film was treated at 220 degrees C and quenched to room temperature, resulting in the formation of a completely amorphous film with a smooth surface. Then, the PHB depolymerases from Pseudomonas stutzeri YM1006 and Ralstonia pickettii T1 were dispersed on the amorphous PLLA thin film. Direct AFM observation has revealed that the PHB depolymerases bind in an elliptic shape on the surface of the PLLA thin film and that a small ridge is created around each enzyme molecule. After removal of the enzymes with 40% ethanol aqueous solution, small hollows were found on the PLLA thin film. These results suggest that a PHB depolymerase interacts with polyester molecules during their adsorption to make a hollow on the substrate surface. PMID- 15360269 TI - Artificial receptor-attached amphiphilic copolymer for barbiturate binding in aqueous media. AB - A water-soluble self-associating amphiphilic copolymer was employed to provide a microenvironment for the solvation of a hydrogen-bonding barbiturate artificial receptor, to facilitate molecular recognition in water. The receptor-attached amphiphilic polymer (RP) was synthesized through random copolymerization of 3% (mol) barbiturate receptor-monomer, 70% (mol) 3-sulfopropyl methacrylate, and 27% (mol) n-dodecyl acrylate. Difference UV spectra of pH 6.5 aqueous solutions of phenobarbital and receptor-polymer (RP) gave peaks and valleys at 272 and 301 nm respectively, consistent with binding characteristics of monomeric barbiturate receptors in chloroform. Specific association between phenobarbital and the receptor-polymer was further indicated based on investigations of a receptor-free control polymer (CP) of similar polar/nonpolar monomer ratio. Micellar electrokinetic chromatography was applied for studying polymer-phenobarbital association, by capillary electrophoresis. PMID- 15360270 TI - Living polypeptides. AB - Block copolypeptides, which combine the self-assembly of block copolymers and the highly ordered 3D structures of proteins, are potential candidates for novel supramolecular structures and biotech applications, such as biosensors, tissue engineering, and selective drug delivery. The synthesis of model block copolypeptides through living nucleophilic/basic polymerization of alpha-amino acid N-carboxyanhydrides (NCAs) has been a challenge for more than fifty years, most probably due to traces of impurities in the system. This problem has been overcome, using high vacuum techniques in order to create and maintain the conditions necessary for the living polymerization of NCAs with primary amines. This method is a general one and opens avenues leading to novel, well-defined polypeptides with various architectures. PMID- 15360271 TI - Resonance energy transfer nanobiosensors based on affinity binding between apo enzyme and its substrate. AB - Systems for glucose monitoring based on resonance energy transfer (RET) and competitive binding using Concanavalin A (Con A) are problematic as a result of problems of toxicity, aggregation, and irreversible binding. This paper presents an improved RET assay wherein Con A was replaced by apo-glucose oxidase (apo GOx). The basic principle for transduction is identical to that used in assays based on Con A-dextran: a reduction in RET from fluorescein isothiocyanate (FITC) to tetramethyl rhodamine isothiocyanate (TRITC) occurs when FITC-dextran (donor) is displaced from TRITC-apo-GOx (acceptor) as a result of the competition of glucose. Fluorescence measurements confirm that the apo-GOx/dextran complexes are highly sensitive to glucose, measured as an increase in the donor peak relative to acceptor due to stepwise addition of glucose. The solution-phase assay showed strong signals and excellent repeatability, with a sensitivity of 0.0163 (ratio units)/mM over the range of 0-90 mM glucose. If properly encapsulated, these sensors can potentially be used for in vivo sensing without the concern of toxicity associated with Con A. PMID- 15360272 TI - Influence of weak and covalent bonds on formation and hydrolysis of gelatin networks. AB - The relative influence of physical and chemical bonds to overall gel properties are explored in gelatin gels. Physical, chemical, chemical-physical, and physical chemical gels are obtained by cooling the protein solution and/or by transglutaminase reaction. Each type of network is characterized by rheology and polarimetry. It is shown that the overall properties as well as the dynamics inside the gels are dependent upon the order of formation and on the relative amount of triple helices and covalent bonds. Enzyme hydrolysis of covalent gels is slower than that of physical gels, as confirmed by the kinetics of helix release and degradation. A scheme is proposed to explain the results at both the physicochemical and the molecular levels. PMID- 15360273 TI - Multilayer biomimetics: reversible covalent stabilization of a nanostructured biofilm. AB - Designed polypeptides and electrostatic layer-by-layer self-assembly form the basis of promising research in bionanotechnology and medicine on development of polyelectrolyte multilayer films (PEMs). We show that PEMs can be formed from oppositely charged 32mers containing several cysteine residues. The polypeptides in PEMs become cross-linked under mild oxidizing conditions. This mimicking of disulfide (S-S) bond stabilization of folded protein structure confers on the PEMs a marked increase in resistance to film disassembly at acidic pH. The reversibility of S-S bond stabilization of PEMs presents further advantages for controlling physical properties of films, coatings, and other applications involving PEMs. PMID- 15360274 TI - Effect of sulfate groups from sulfuric acid hydrolysis on the thermal degradation behavior of bacterial cellulose. AB - When used as fillers in polymer composites, the thermostability of cellulose crystals is important. Sulfate groups, introduced during hydrolysis with sulfuric acid, are suspected to diminish the thermostability. To elucidate the relationship between the hydrolysis conditions, the number of sulfate groups introduced, and the thermal degradation behavior of cellulose crystals, bacterial cellulose was hydrolyzed with sulfuric acid under different hydrolysis conditions. The number of sulfate groups in the crystals was determined by potentiometric titration. The thermal degradation behavior was investigated by thermogravimetric analysis. The sulfate group content increased with acid concentration, acid-to-cellulose ratio, and hydrolysis time. Even at low levels, the sulfate groups caused a significant decrease in degradation temperatures and an increase in char fraction confirming that the sulfate groups act as flame retardants. Profile analysis of the derivative thermogravimetric curves indicated thermal separation of the degradation reactions by the sulfate groups into low- and high-temperature processes. The Broido method was used to determine activation energies for the degradation processes. The activation energies were lower at larger amounts of sulfate groups suggesting a catalytic effect on the degradation reactions. For high thermostability in the crystals, low acid concentrations, small acid-to-cellulose ratios, and short hydrolysis times should be used. PMID- 15360275 TI - Lignin incorporation combined with electron-beam irradiation improves the surface water resistance of starch films. AB - We investigated the potential of an electron-beam post-treatment to tailor the properties of 70/30 and 80/20 wt. extruded starch-lignin films. The effect of a 400 kGy radiation on films differing essentially by the kind of lignins incorporated (lignosulfonates/alkali lignins) was assessed both at the macroscopic and the molecular levels. Changes in the polymer molecular structure were studied by IR spectroscopy, by thioacidolysis as well as by model compound experiments. Electron beam-irradiation at 400 kGy, a rather high dose for processing natural polymers, alters to some extent the mechanical resistance of the starch-based materials. However this treatment substantially reduces the hydrophilic surface properties of the films, while not harming their biodegradability. Involved in radical cross-coupling reactions, lignin phenolic compounds are likely to play a primary role in the formation of a hydrophobic condensed network. This study suggests that lower irradiation doses might yield biomaterials with improved usage properties. PMID- 15360276 TI - Biodegradation of aliphatic homopolyesters and aliphatic-aromatic copolyesters by anaerobic microorganisms. AB - The anaerobic degradability of natural and synthetic polyesters is investigated applying microbial consortia (3 sludges, 1 sediment) as well as individual strains isolated for this purpose. In contrast to aerobic conditions, the natural homopolyester polyhydroxybutyrate (PHB) degrades faster than the copolyester poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV). For the synthetic polyester poly(epsilon-caroplacton) (PCL), microbial degradation in the absence of oxygen could be clearly demonstrated; however, the degradation rate is significantly lower than for PHB and PHBV. Other synthetic polyesters such as poly(trimethylene adipate) (SP3/6), poly(tetramethylene adipate) (SP4/6), and aliphatic-aromatic copolyesters from 1,4-butanediol, terephthalic acid, and adipic acid (BTA copolymers) exhibit only very low anaerobic microbial susceptibility. A copolyester with high amount of terephthalic acid (BTA 40:60) resisted the anaerobic breakdown even under thermophilic conditions and/or when blended with starch. For the synthetic polymers, a number of individual anaerobic strain could be isolated which are able to depolymerize the polymers and selected strains where identified as new species of the genus Clostridium or Propionispora. Their distinguished degradation patterns point to the involvement of different degrading enzymes which are specialized to depolymerize either the natural polyhydroxyalkanoates (e.g., PHB), the synthetic polyester PCL, or other synthetic aliphatic polyesters such as SP3/6. It can be supposed that these enzymes exhibit comparable characteristics as those described to be responsible for aerobic polyester degradation (lipases, cutinases, and PHB-depolymerases). PMID- 15360277 TI - Solvent removal during synthetic and Nephila fiber spinning. AB - The process by which spiders make their mechanically superior fiber involves removal of solvent (water) from a concentrated protein solution while the solution flows through a progressively narrowing spinning canal. Our aim was to determine a possible mechanism of spider water removal by using a computational model. To develop appropriate computational techniques for modeling of solvent removal during fiber spinning, a study was first performed using a synthetic solution. In particular, the effect of solvent removal during elongational flow (also exhibited in the spinning canal of the spider) on fiber mechanical properties was examined. The study establishes a model for solvent removal during dry spinning of synthetic fibers, assuming that internal diffusion governs solvent removal and that convective resistance is small. A variable internal solvent diffusion coefficient, dependent on solvent concentration, is also taken into account in the model. An experimental setup for dry (air) spinning was used to make fibers whose diameter was on the order of those made by spiders (approximately 1 microm). Two fibers of different thickness, corresponding to different spinning conditions, were numerically modeled for solvent removal and then mechanically tested. These tests showed that the thinner fiber, which lost more solvent under elongational flow, had 5-fold better mechanical properties (elastic modulus of 100 MPa and toughness of 15 MJ/m3) than the thicker fiber. Even though the mechanical properties were far from those of dragline spider silk (modulus of 10 GPa and toughness of 150 MJ/m3), the experimental methodology and numerical principles developed for the synthetic case proved to be valuable when establishing a model for the Nephila spinning process. In this model, an assumption of rapid convective water removal at the spinning canal wall was made, with internal diffusion of water through the fiber as the governing process. Then the diffusion coefficient of water through the initial spinning solution, obtained ex vivo from the Nephila clavipes major ampullate gland, was determined and incorporated into the numerical procedure, along with the wall boundary conditions and canal geometry. Also, a typical fiber reeling speed during web making, as well as the assumption of a dry exiting fiber, were included in the model. The results show that a cross-section of spinning solution (dope), which is initially 70% water, spends 19 s in the spinning canal in order to emerge dry. While the dope cross-section traverses the canal, its velocity increases from 0.37 mm/s at the entrance to 12.5 mm/s at the canal exit. The obtained results thus indicate that simple diffusion, along with the dry wall boundary condition, is a viable mechanism for water removal during typical Nephila fiber spinning. PMID- 15360278 TI - Micropatterned carbohydrate displays by self-assembly of glycoconjugate polymers on hydrophobic templates on silicon. AB - We report a novel strategy for micropatterned carbohydrate displays on Si substrates. This method exploited the hydrophobic-hydrophilic microfabrication by photolithography of ODS-SAM on Si substrates and the subsequent selective self assembly of glycoconjugate polymers onto the hydrophobic regions. Protein micropatterning by molecular recognition on the carbohydrate substrates was also successful. PMID- 15360279 TI - Targeted worm micelles. AB - Giant and stable worm micelles formed from poly(ethylene glycol) (PEG)-based diblock copolymer amphiphiles have the potential advantage compared to smaller assemblies for delivery of a large quantity of hydrophobic drugs or dyes per carrier. Here we show that worm micelles can be targeted to cells with internalization and delivery of nontoxic dyes as well as cytotoxic drugs. Constituent copolymers are end-biotinylated to mediate high affinity binding of worm micelles to both avidin-bearing surfaces and biotin-specific receptors on smooth muscle cells. Pristine worm micelles, that lack biotin, show much less frequent and nonspecific point attachments to the same surfaces. Biotinylated worm micelles prove stable in aqueous solution for at least a month and also prove capable of loading, retaining, and delivering hydrophobic dyes and drugs. The results thus demonstrate the feasibility of targeted delivery by polymeric worm micelles. PMID- 15360280 TI - Controlling rigidity and degradation of alginate hydrogels via molecular weight distribution. AB - The mechanical rigidity and degradation rate of hydrogels utilized as cell transplantation vehicles have been regarded as critical factors in new tissue formation. However, conventional approaches to accelerate the degradation rate of gels deteriorate their function as a mechanical support in parallel. We hypothesized that adjusting the molecular weight distribution of polymers that are hydrolytically labile but capable of forming gels would allow one to alter the degradation rate of the gels over a broad range, while limiting the range of their elastic moduli (E). We investigated this hypothesis with binary alginate hydrogels formed from both ionically and covalently cross-linked partially oxidized (1% uronic acid residues), low [molecular weight (MW) approximately 60,000 g/mol] and high MW alginates (MW approximately 120,000 g/mol) in order to examine the utility of this approach with various cross-linking strategies. Increasing the fraction of low MW alginates to 0.50 maintained a value of E similar to that for the high MW alginate gels but led to faster degradation, irrespective of the cross-linking mode. This result was attributed to a faster separation between cross-linked domains upon chain breakages for the low MW alginates, coupled with their faster chain scission than the high MW alginates. The more rapidly degrading oxidized binary hydrogels facilitated the formation of new bone tissues from transplanted bone marrow stromal cells, as compared with the nonoxidized high MW hydrogels. The results of these studies will be useful for controlling the physical properties of a broad array of hydrogel-forming polymers. PMID- 15360281 TI - Structural changes and aggregation of human influenza virus. AB - The pH-induced change in the structure and aggregation state of the PR-8 and X-31 strains of intact human influenza virus has been studied in vitro. Reducing the pH from 7.4 to 5.0 produces a large increase in the intensity of light scattered to low angles. A modest increase in the polydispersity parameter from cumulants fits to the dynamic light scattering correlograms accompanies the increase, as does a change in how that parameter varies with scattering angle. These trends imply that the virus particles are not uniform, even at pH 7.4, and tend to aggregate as pH is reduced. The scattering profiles (angular dependence of intensity) never match those of isolated, spherical particles of uniform size, but the deviations from that simple model remain modest at pH 7.4. At pH 5.0, scattering profiles calculated for aggregates of uniformly sized spheres come much closer to matching the experimental data than those computed for isolated particles. Although these observations indicate that acid-induced aggregation develops over a period of minutes to hours after acidification, a nearly instantaneous increase in hydrodynamic size is the first response of intact virus particles to lower pH. PMID- 15360282 TI - Enhanced stability of core-surface cross-linked micelles fabricated from amphiphilic brush copolymers. AB - "Stealth" nanoparticles made from polymer micelles have been widely explored as drug carriers for targeted drug delivery. High stability (i.e., low critical micelle concentration (CMC)) is required for their intravenous applications. Herein, we present a "core-surface cross-linking" concept to greatly enhance nanoparticle's stability: amphiphilic brush copolymers form core-surface cross linked micelles (nanoparticles) (SCNs). The amphiphilic brush copolymers consisted of hydrophobic poly(epsilon-caprolactone) (PCL) and hydrophilic poly(ethylene glycol) (PEG) or poly(2-(N,N-dimethylamino)ethyl methacrylate) (PDMA) chains were synthesized by macromonomer copolymerization method and used to demonstrate this concept. The resulting SCNs were about 100 times more stable than micelles from corresponding amphiphilic block copolymers. The size and surface properties of the SCNs could be easily tailored by the copolymer's compositions. PMID- 15360283 TI - Micropatterning of nanoengineered surfaces to study neuronal cell attachment in vitro. AB - Methods for producing protein patterns with defined spatial arrangement and micro and nanoscale features are important for studying cellular-level interactions, including basic cell-cell communications, cell signaling, and mechanisms of drug action. Toward this end, a straightforward, versatile procedure for fabricating micropatterns of bioactive nanofilm coatings as multifunctional biological testbeds is demonstrated. The method, based on a combination of photolithography and layer-by-layer self-assembly (LbL), allows for precise construction of nanocomposite films of potentially complex architecture, and patterning of these films on substrates using a modified lift-off (LO) procedure. As a first step in evaluating nanostructures made with this process, "comparison chips," comprising two coexisting regions of square patterns with relevant proteins/polypeptides on a single substrate, were fabricated with poly(diallyldimethylammonium chloride) (PDDA) as a cell-repellent background. Using neuronal cells as a model biological system, comparison chips were produced with secreted phospholipase A2 (sPLA2), a known membrane-active enzyme for neurons, for direct comparison with gelatin, poly-l-lysine (PLL), or bovine serum albumin (BSA). Fluorescence microscopy, surface profilometry, and atomic force microscopy techniques were used to evaluate the structural properties of the patterns on these chips and show that the patterning technique was successful. Preliminary cell culture studies show that neurons respond and bind specifically to the sPLA2 enzyme embedded in the polyelectrolyte thin films and present as the outermost layer. These findings point to the potential for this method to be applied in developing test substrates for a broad array of studies aimed at identifying important biological structure-function relationships. PMID- 15360284 TI - Degradation behavior of poly(epsilon-caprolactone)-b-poly(ethylene glycol)-b poly(epsilon-caprolactone) micelles in aqueous solution. AB - Poly(epsilon-caprolactone)-b-poly(ethylene glycol)-b-poly(epsilon-caprolactone) triblock copolymers were synthesized by the ring-opening polymerization of epsilon-caprolactone in the presence of hydroxyl-terminated poly(ethylene glycol) with different molecular weights, using stannous octoate catalyst. Micelles prepared by the precipitation method with these triblock copolymers exhibit a core-shell structure. The degradation behaviors of these core-shell micelles in aqueous solution were investigated by FT-IR, 1H NMR, GPC, DLS, TEM, and AFM. It was found that the degradation behavior of micelles in aqueous solution was quite different from that of bulk materials. The size of the micelles increased in the initial degradation stages and decreased gradually when the degradation period was extended. The caprolactone/ethylene oxide (CL/EO) ratio in micelles measured by NMR also shows an increase at the initial degradation stage and a decrease at later stages. The morphology of these micelles became more and more irregular during the degradation period. We explain the observed behavior by a two-stage degradation mechanism with interfacial erosion between the cores and the shells followed by core erosion. PMID- 15360285 TI - Structural analysis of Bombyx mori silk fibroin peptides with formic acid treatment using high-resolution solid-state 13C NMR spectroscopy. AB - Bombyx mori silk fibroin fiber is a fibrous protein produced by the silkworm at room temperature and from an aqueous solution whose primary structure is highly repetitive. In this study we analyzed the structural characteristics of native peptides, derived from B. mori silk fibroin, with formic acid treatment using high-resolution solid-state 13C NMR. We establish that the Ser residue bearing a short polar side chain has the ability to stabilize the conformation formed in the model peptides due to its ability to form intermolecular hydrogen bonds involving its hydroxyl group as a donor and the carbonyl groups of other residues as acceptors. On the other hand, insertion of Tyr residue in the basic (AG)n and (AGSGAG)n sequence motifs usually exhibited disruptive effects on the preferred conformations. Moreover, the environmental effect was investigated by mixing the native Cp fraction with the model peptides, showing that there is no significant structural difference on the Ser-containing peptides, while structural transformation was observed on the peptides containing the GAAS unit. This may be attributed to the fact that the Cp fraction promotes the formation of an antiparallel beta-sheet in the Ala-Ala unit. Such periodically disrupted ordered structures in the semicrystalline region of B. mori silk fibroin may be critical not only for facilitating the conformational transformation from silk I to silk II structural form but also for having some correlation with the unique properties of the silk materials. PMID- 15360286 TI - Photon-modulated changes of cell attachments on poly(spiropyran-co-methyl methacrylate) membranes. AB - Spiropyran is a photoresponsive molecule, and nonionic spiropyran is reversibly changed by UV irradiation to a hydrophilic polar, zwitterionic merocyanine isomer, and back again by visible light irradiation. A copolymer of nitrobenzospiropyran and methyl methacrylate, poly(NSP-co-MMA) was used as a material with a photosensitive surface. UV irradiation of the photosensitive surface of poly(NSP-co-MMA)-coated glass plates decreased the water contact angles (11 +/- 1 degrees ) and increased diameter of a water drop relative to the unexposed surface. Light-induced detachment of platelets and mesenchymal stem (KUSA-A1) cells on poly(NSP-co-MMA)-coated glass plates was observed upon simple- and patterned-light irradiation, whereas no light-induced detachment of platelets and mesenchymal stem cells was observed on poly(methyl methacrylate)-coated glass plates. This is a result of the change from a closed nonpolar spiropyran to the polar zwitterionic merocyanine isomer induced by UV irradiation. Light-induced detachment of fibrinogen adsorbed on poly(NSP-co-MMA) coated glass plates was also observed in this investigation. PMID- 15360287 TI - Amylopectin molecular structure reflected in macromolecular organization of granular starch. AB - For lintners with negligible amylose retrogradation, crystallinity related inversely to starch amylose content and, irrespective of starch source, incomplete removal of amorphous material was shown. The latter was more pronounced for B-type than for A-type starches. The two predominant lintner populations, with modal degrees of polymerization (DP) of 13-15 and 23-27, were best resolved for amylose-deficient and A-type starches. Results indicate a more specific hydrolysis of amorphous lamellae in such starches. Small-angle X-ray scattering showed a more intense 9-nm scattering peak for native amylose deficient A-type starches than for their regular or B-type analogues. The experimental evidence indicates a lower contrasting density within the "crystalline" shells of the latter starches. A higher density in the amorphous lamellae, envisaged by the lamellar helical model, explains the relative acid resistance of linear amylopectin chains with DP > 20, observed in lintners of B type starches. Because amylopectin chain length distributions were similar for regular and amylose-deficient starches of the same crystal type, we deduce that the more dense (and ordered) packing of double helices into lamellar structures in amylose-deficient starches is due to a different amylopectin branching pattern. PMID- 15360288 TI - Morphology and enzymatic degradation of oriented thin film of ultrahigh molecular weight poly[(R)-3-hydroxybutyrate]. AB - Thin films of ultrahigh molecular weight poly[(R)-3-hydroxybutyrate] (P(3HB)) were sheared and isothermally crystallized at 100 degrees C. Transmission electron microscopy and atomic force microscopy (AFM) observations revealed that thick fibrous textures, on which lamellae are overgrown normal to the long axis of the fibril, run parallel to the shearing direction. A selected area electron diffraction pattern taken from the fibrils exhibits a fiber pattern of P(3HB) alpha-modification, and the crystallographic c-axis (chain axis) of P(3HB) is set parallel to the long axis of the fibril. In situ AFM observations of enzymatic degradation for the thin film were performed with an extracellular P(3HB) depolymerase from Ralstonia pickettii T1 in a buffer solution. The film surface and thickness became rougher and thinner, respectively, with time after adding the enzyme. During the degradation, fine shish-kebab structures appeared gradually. This fact supports that the amorphous region in the film is preferentially degraded rather than the crystalline one by the depolymerase. The in situ AFM observations also revealed that one thick fibril in the original film is composed of three different states, namely, finer fibril (shish), stacked lamellae (kebab) in edge-on state, and the surrounding amorphous phase. PMID- 15360289 TI - Split crystallization during debranching of maltodextrins at high concentration by isoamylase. AB - Debranching and crystallization occurring during the enzymatic treatment of 25% (w/v) aqueous solutions of maltodextrins by isoamylase at 52 degrees C were studied. The morphology as well as the crystal and molecular structures of the precipitates formed at different stages of the reaction were characterized. Two types of resulting products, differing in terms of structure and morphology, were evidenced. A loose B-type network, containing linear and branched chains of highest molecular weight, was mainly formed during the first 12 h of reaction, whereas aggregates of A-type lamellar crystals, made of short linear chains, were predominantly obtained between 12 and 48 h. The aggregation behavior as a function of temperature and molecular weight distribution of such substrates was discussed and compared to that of related starch products. PMID- 15360290 TI - Facilitated synthesis of inulin esters by transesterification. AB - Inulin, the polydisperse polyfructose, extracted from chicory, has been modified via transesterification, using fatty acid methyl esters (FAME). The grafting of an alkyl chain onto the inulin backbone under different conditions for the development of potential tensio-active derivatives is described. The modification of the biopolymer was performed in polar organic solvents, such as dimethyl sulfoxide (DMSO) and N-methylpyrrolidinone (NMP). Depending on the type of solvent, different catalytic systems, such as DMSO-Na+, NaH, and NaOMe, were used and compared in reaction efficiency and reproducibility. Therefore the synthesized derivatives were characterized by 1H- and 13C NMR. The methods using NaH had a mean reaction efficiency of 80%, whereas the one using NaOMe showed a slight decrease in reaction efficiency to 75%. However, the method using NaOMe in NMP proved to be the preferred way to graft the inulin backbone with FAME on a bigger scale. The methods using DMSO as a solvent were not attractive since the end products had a specific bad smell. PMID- 15360291 TI - Photoresponsive nanogels formed by the self-assembly of spiropyrane-bearing pullulan that act as artificial molecular chaperones. AB - Novel photoresponsive nanogels were prepared by the self-assembly of spiropyrane bearing pullulan (SpP). The solution properties of the nanogels could be controlled by photostimulation via isomerization between hydrophobic spiropyrane and hydrophilic merocyanine. The molecular chaperone-like activity of the nanogels in protein refolding was investigated. The activity of citrate synthase significantly increased when the amphiphilicity of SpP nanogels was switched by photostimulation. PMID- 15360292 TI - Synthesis and characterization of biodegradable poly(ethylene glycol)-block poly(5-benzyloxy-trimethylene carbonate) copolymers for drug delivery. AB - Amphiphilic diblock copolymers with various block compositions were synthesized with monomethoxy-terminated poly(ethylene glycol) (MePEG) as the hydrophilic block and poly(5-benzyloxy-trimethylene carbonate) (PBTMC) as the hydrophobic block. When the copolymerization was conducted using MePEG as a macroinitiator and stannous 2-ethylhexanoate (Sn(Oct)2) as a catalyst, the molecular weight of the second block was uncontrollable, and the method only afforded a mixture of homopolymer and copolymer with a broad molecular weight distribution. By contrast, the use of the triethylaluminum-MePEG initiator yielded block copolymers with controllable molecular weight and a more narrow molecular weight distribution than the copolymers obtained using Sn(Oct)2. GPC and 1H NMR studies confirmed that the macroinitiator was consumed and the copolymer composition was as predicted. Two of the newly synthesized MePEG-b-PBTMC copolymers were evaluated in terms of properties primarily relating to their use in micellar drug delivery. MePEG-b-PBTMC micelles with a narrow monomodal size distribution were prepared using a high-pressure extrusion technique. The MePEG-b-PBTMC copolymers were also confirmed to be biodegradable and noncytotoxic. PMID- 15360293 TI - Amyloid fibril formation by peptide LYS (11-36) in aqueous trifluoroethanol. AB - Peptide LYS (11-36), derived from the beta-sheet region of T4 lysozyme, forms an amyloid fibril in aqueous trifluoroethanol (TFE) at elevated temperature. The peptide has a moderate alpha-helix content in 20 and 50% (v/v) TFE solution; large quantities of fibrils were formed after incubation at 55 degrees C for 2 weeks as monitored by a thioflavin T fluorescence assay. No fibrils were observed when the peptide initially existed predominantly as a random coil or as a complete alpha helix. Our results suggest that a moderate amount of alpha helix and random coil present in the peptide initially facilitates the fibril-formation process, but a high alpha-helix content inhibits fibril formation. Transmission electron microscopy revealed several types of fibril morphologies at different TFE concentrations. The fibrils were highly twisted and consisted of interleaved protofilaments in 50% TFE, while smooth and flat ribbonlike fibrils were found in 20% TFE. In 50% TFE, the fibril growth rate of LYS (11-36) was found to depend strongly on peptide concentration and seeding but was insensitive to solution pH and ionic strength. PMID- 15360294 TI - 1H magnetization transfer in hydrated gluten and flour: effects of wheat aging. AB - The interaction of water with flour or gluten in hydrated samples was investigated by proton magnetization transfer measurements. Flour and gluten from both durum and bread wheat seeds, either unaged or artificially aged over different periods of time, were investigated. Measurements were performed at several radio frequency power levels and frequency offsets, and the data were quantitatively modeled by two interacting pools, a liquid (water) and a solid (macromolecules) one. A super-Lorentzian line shape well described the magnetization of the solid pool. Magnetization transfer was found to be more efficient for flour with respect to gluten samples, in agreement with their hydrophilic/hydrophobic behavior. The aging treatment of seeds resulted in a minor degree of interaction between macromolecules and water. PMID- 15360295 TI - Ethyl oleate synthesis using Candida rugosa lipase in a solvent-free system. Role of hydrophobic interactions. AB - The solvent-free esterification reaction of a commercial oleic acid and ethanol was selected as the test reaction for Candida rugosa lipase immobilized on polypropylene (PP) at 318 K (initial molar ratio 1:1). Adding of water from 0 to 30 wt. % (in gram per gram of fatty acid x 100) and the pretreatment of Candida rugosa lipase with polyethylenglycol (PEG), octane, and acetone increases the conversion to ethyl esters. The role of hydrophobic interactions of the lipase with PP and PEG was studied using molecular mechanics (MM2) for calculation of steric energies and the parametrized model (PM3) for calculation of enthalpy changes upon interaction. The nonpolar lateral groups of amino acids interact strongly with PP, whereas polar groups interact more strongly with PEG. Both interactions stabilize the open, active conformation of the lipase from Candida rugosa. Activities ranged from 5 x 10(-5) to 2.0 x 10(-4) mol ethyl oleate/h/mg enzyme, depending on reaction conditions. Steric energy changes vary between +30 and -10 kcal/mol, whereas the enthalpy changes ranged from +10 to -10 kcal/mol. PMID- 15360296 TI - Hydrolysis of poly(alkylene amidophosphate)s containing amino acid or peptide residues in the side groups. Kinetics and selectivity of hydrolysis. AB - Kinetics of hydrolysis of poly(alkylene amidophosphate)s with amino acids or dipeptides as the side groups was studied by 31P NMR at pH 1.5, 6.5, and 8.5. The direction of hydrolysis and the relative rate coefficients of breaking P-O bonds in the main chain and P-N bonds in the side groups depend strongly on the pH of the medium of hydrolysis. The P-N (amide) bond hydrolyzes much faster than the P O (ester) bond in acidic and close to neutral conditions (negligible P-O hydrolysis), whereas above pH > or = 8.5 these differences are much smaller. For instance, for 4-Ala the rate coefficients of hydrolysis are equal (in H2O at 37 degrees C and pH 8.5) to 1.9 x 10(-8) s(-1) and 1.0 x 10(-9) s(-1) for the P-N and P-O bonds, respectively, quite different from the values found for the low molecular model 2 (0 and 1.4 x 10(-7) s(-1), respectively). PMID- 15360297 TI - Acid extraction and purification of recombinant spider silk proteins. AB - A procedure has been developed for the isolation of recombinant spider silk proteins based upon their unique stability and solubilization characteristics. Three recombinant silk proteins, (SpI)7, NcDS, and [(SpI)4/(SpII)1]4, were purified by extraction with organic acids followed by affinity or ion exchange chromatography resulting in 90-95% pure silk solutions. The protein yield of NcDS (15 mg/L culture) and (SpI)7 (35 mg/L) increased 4- and 5-fold, respectively, from previously reported values presumably due to a more complete solubilization of the expressed recombinant protein. [(SpI)4/(SpII)1]4, a hybrid protein based on the repeat sequences of spidroin I and spidroin II, had a yield of 12.4 mg/L. This method is an effective, reproducible technique that has broad applicability for a variety of silk proteins as well as other acid stable biopolymers. PMID- 15360298 TI - Complexation of DNA with poly(methacryl oxyethyl trimethylammonium chloride) and Its poly(oxyethylene) grafted analogue. AB - Intermolecular complexes of genomic polydisperse DNA with synthetic polycations have been studied. Two cationic polymers have been used, a homopolymer poly(methacryl oxyethyl trimethylammonium chloride) (PMOTAC) and its analogue grafted with poly(oxyethylene). The amount of poly(oxyethylene) grafts in the copolymer was 15 mol % and Mw of the graft was 200 g/mol. Salmon DNA (sodium salt) was used. The average molecular weight (Mw) of DNA was 10.4 x 10(6) g/mol. Conductivity, pH, and dynamic light scattering studies were used to characterize the complexes. The size and shape of the polyelectrolyte complex particles have been studied as a function of the cation-to-anion ratio in aqueous solutions of varying ionic strengths. The polyelectrolyte complexes have extremely narrow size distributions taking into account the polydispersity of the polyelectrolytes studied. The poly(oxyethylene) grafts on PMOTAC promote the formation of small colloidally stabile complex particles. Addition of salt shifts the macroscopic phase separation toward lower polycation content; that is, complexes partly phase separate with the mixing ratios far from 1:1. Further addition of salt to the turbid, partly phase separated solution results in the dissociation of complexes and the polycation and DNA dissolve as individual chains. PMID- 15360299 TI - Cascade synthesis of chiral block copolymers combining lipase catalyzed ring opening polymerization and atom transfer radical polymerization. AB - The enantioselective polymerization of methyl-substituted epsilon-caprolactones using Novozym 435 as the catalyst was investigated. All substituted monomers could be polymerized except 6-methyl-epsilon-caprolactone (6-MeCL), which failed to propagate after ring opening. Interestingly, an odd-even effect in the enantiopreference of differently substituted monomers was observed. The combination of 4-methyl-epsilon-caprolactone with Novozym 435 showed good enantioselectivity also in bulk polymerization and resulted in enantiomerically enriched P((S)-4-MeCL) (eep up to 0.88). Subsequently, a novel initiator combining a primary alcohol to initiate the ring opening polymerization and a tertiary bromide to initiate atom transfer controlled radical polymerization (ATRP) was synthesized, and showed high initiator efficiencies (> 90%) in the ring opening polymerization of 4-methyl-epsilon-caprolactone in bulk. In addition, the enantioselectivity was retained (E = 11). By using Ni(PPh3)2Br2 as the ATRP catalyst, Novozym 435 could be effectively inhibited at the desired conversion of 4-methyl-epsilon-caprolactone, thus ensuring a high enantiomeric excess in the polymer backbone. At the same time, Ni(PPh3)2Br2 catalyzed the ATRP of methyl methacrylate resulting in the formation of P((S)-4-MeCL-b-MMA) block copolymers. By this combination of two inherently different polymerization reactions, chiral P((S)-4-MeCL-b-MMA) block copolymers can be conveniently obtained in one pot without intermediate workup. PMID- 15360300 TI - Enzymatic copolymerization alters the structure of unpolymerized mixtures of the biomimetic monomers: the amphiphilic decyl ester of L-tyrosine and L tyrosineamide--an AFM investigation of nano- to micrometer-scale structure differences. AB - Previously, we have shown that the amphiphilic decyl esters of both D- and L tyrosine (DELT) self-assemble in aqueous solution above their critical micelle concentration values to form long rodlike structures that can be enzymatically polymerized. In the current study, we have examined the self-assembled structures of unpolymerized and enzymatically (horseradish peroxidase) copolymerized 1:1 molar mixtures of DELT with the nonamphiphilic comonomer L-tyrosineamide. The structures were examined following adsorption to gold-coated mica surfaces using optical microscopy and scanning electron microscopy, but primarily noncontact atomic force microscopy. Both unpolymerized and copolymerized 1:1 comonomer mixture aggregates produced amorphous to spherical shaped structures, exhibiting increased flexibility that contrasted with our previous observations of the more highly ordered long rodlike structures seen with the pure DELT. The unpolymerized comonomer aggregates were amorphous and of varying size. Interestingly, they contained occasional novel structures-smooth, sharp, nipplelike features that rose hundreds of nanometers above the smooth aggregate surface. However, upon enzymatic copolymerization, the structures are altered, forming nearly hemispherical aggregates in contact with each other on the surface. These structures possessed diameters of 1.51 +/- 0.24 microm. The copolymerized structures lacked any evidence of the sharp nipplelike features observed in the unpolymerized sample, but they did exhibit nanometer-scale detailed surface features, indicative of a higher degree of internal organization. The measured surface roughness of the copolymerized comonomer mixture was more than 10 times greater than the surface roughness of the unpolymerized comonomer mixture. PMID- 15360301 TI - Heparin conjugated polylactide as a blood compatible material. AB - A heparin-conjugated biodegradable polymer (PLA-heparin) by the direct coupling of heparin to polylactide (PLA) was synthesized and characterized. The surface exposed heparin content associated PLA-heparin was measured to be 0.067 microg/cm2. PLA-heparin coated surface has shown higher hydrophilicity rather than control PLA surface. The clotting time of PLA-heparin conjugate measured by activated partial thromboplastin time (APTT) was significantly prolonged as compared to PLA. The bioactivity of bound heparin measured by APTT corresponds to 17.4% of free heparin. It has been also demonstrated that the conjugation of heparin suppresses the protein adsorption as well as the platelet adhesion. These results indicate that the unique property of bound heparin has an inhibiting influence on the coagulation, plasma protein adsorption, and subsequent platelet adhesion systems. This novel PLA-heparin conjugate could be applied as blood/tissue compatible biodegradable materials for implantable medical devices and tissue engineering. PMID- 15360302 TI - Versatile and efficient formation of colloids of biopolymer-based polyelectrolyte complexes. AB - The formation of colloids based on polyelectrolyte complexes (PECs) of biopolymers was investigated through the complexation between two charged polysaccharides, chitosan as polycation, and dextran sulfate as polyanion. The slow dropwise addition of components, generally used for the formation of PECs, allowed to elaborate both cationic or anionic particles with an excess of chitosan or dextran sulfate, respectively. The PEC particles featured a core/shell structure, the hydrophobic core resulting from the segregation of complexed segments whereas excess component in the outer shell ensured the colloidal stabilization against further coagulation. Considering the host/guest concept for the formation of PECs, the influence of the molecular weight of components on particles sizes could be well explained by the chain length ratios of the two polymers. As an irreversible flocculation occurred with a dropwise approach for both cationic and anionic PEC particles when the mixing ratio was close to unity, a more versatile, and simpler to setup, method was designed: the one-shot addition of one solution to the other. Because process of addition is faster than the flocculation, cationic or anionic particles could be elaborated irrespective of the order of addition of the reactant. Characterization of these particles by quasielastic light scattering, electrophoresis, and scanning electron microscopy revealed very similar properties to those obtained by a slow dropwise approach. Critical coagulation concentrations of 0.12 and 0.09 M (with sodium chloride) for cationic and anionic particles evidenced a mostly electrostatic stabilization. PMID- 15360303 TI - Collapse and association of denatured lentinan in water/dimethlysulfoxide solutions. AB - Triple helical lentinan, beta-(1-->3)-D-glucan from Lentinus edodes, was denatured in dimethlysulfoxide (DMSO) into single random coils. The DMSO solutions of randomly coiled lentinan were diluted with pure water to different wH (the weight fraction of water in the mixed solvent), and their specific optical rotation [alpha]D, reduced viscosity (lnetar)/c, and hydrodynamic radius Rh were investigated as a function of wH and storage time t. With an increase of wH from 0.1 to 0.2, [alpha]D increased sharply, suggesting that transition of conformation of the macromolecules has occurred. When wH was lower than 0.1, (lnetar)/c of lentinan in water-diluted DMSO exhibited the almost same value as that in pure DMSO and changed hardly with increasing t. Interestingly, (lnetar)/c decreased to reach a minimum with a further increase of wH from 0.1 to 0.25 and then increased with a continuous increase of wH from 0.25 to 0.5. Both (lnetar)/c and Rh of the denatured lentinan in water-diluted DMSO with wH of approximately 0.25 both exhibited a minimum, indicating that collapsed coil chains have occurred. All of the experimental findings revealed that the behaviors of lentinan in water-diluted DMSO solution with wH < 0.1 were consistent with that in good solvent, DMSO. When wH = 0.25, the quality of the mixed solvents became worse, and the dominant intramacromolecular hydrogen-bond interaction enhanced, leading to minimum of viscosity and size of the chains as a result of the collapsed coils. When wH > 0.25, the quality of the mixture weakens further, and the intermolecular hydrogen-bond interaction enhanced and was dominant, leading to aggregation of the collapsed chains. PMID- 15360304 TI - Comparative study of the second and third heterogeneous deacetylations of alpha- and beta-chitins in a multistep process. AB - Second and third heterogeneous deacetylations in a multistep process under argon atmosphere of alpha- and beta-chitins in the presence of 50% (w/v) NaOH, for temperatures ranging from 80 to 110 degrees C, were comparatively studied in order to optimize the multistep process of deacetylation. Along with the successive reactions, we observed important changes of chemical behavior with the crystalline state related to alpha- and beta-chitins, amorphous and partially deacetylated chitin, and chitosan. Thanks to the full reacetylation of all the deacetylated samples, we succeeded in estimating the oxidoreductive alkaline degradation occurring during deacetylation, whatever the degree of acetylation (DA) of the copolymer. It clearly appeared that the crystalline state of the samples was the key parameter on which depended the rate constants of both alkaline hydrolysis and deacetylation and, consequently, the activation energy Ea and the preexponential factor A. We may now propose optimal conditions allowing the production of well-defined chitosans with low DAs and higher molecular weights than those usually reported in the literature. PMID- 15360305 TI - Elasticity of native and cross-linked polyelectrolyte multilayer films. AB - Mechanical properties of polyelectrolyte multilayer films were studied by nanoindentation using the atomic force microscope (AFM). Force-distance measurements using colloidal probe tips were systematically obtained for supported films of poly(L-lysine) and hyaluronan that are suited to bio application. Both native and covalently cross-linked films were studied as a function of increasing layer number, which increases film thickness. The effective Young's modulus perpendicular to the film, Eperpendicular, was determined to be a function of film thickness, cross-linking, and sample age. Thick PEM films exhibited a lower Eperpendicular than thinner PEM, whereas the Young's modulus of cross-linked films was more than 10-fold larger than native films. Moduli range from approximately 20 kPa for native films up to approximately 800 kPa for cross-linked ones. Young's moduli increased slightly with sample age, plateauing after approximately 4 weeks. Spreading of smooth muscle cells on these substrates with pre-attached collagen proved to be highly dependent on film rigidity with stiffer films giving greater cell spreading. PMID- 15360306 TI - Novel method using a temperature-sensitive polymer (methylcellulose) to thermally gel aqueous alginate as a pH-sensitive hydrogel. AB - A novel method using a temperature-sensitive polymer (methylcellulose) to thermally gel aqueous alginate blended with distinct salts (CaCl2, Na2HPO4, or NaCl), as a pH-sensitive hydrogel was developed for protein drug delivery. It was noted that the salts blended in hydrogels may affect the structures of an entangled network of methylcellulose and alginate and have an effect on their swelling characteristics. The methylcellulose/alginate hydrogel blended with 0.7 M NaCl (with a gelation temperature of 32 degrees C) demonstrated excellent pH sensitivity and was selected for the study of release profiles of a model protein drug (bovine serum albumin, BSA). In the preparation of drug-loaded hydrogels, BSA was well-mixed to the dissolved aqueous methylcellulose/alginate blended with salts at 4 degrees C and then gelled by elevating the temperature to 37 degrees C. This drug-loading procedure in aqueous environment at low temperature may minimize degradation of the protein drug while achieving a high loading efficiency (95-98%). The amount of BSA released from test hydrogels was a function of the amount of alginate used in the hydrogels. The amount of BSA released at pH 1.2 from the test hydrogel with 2.5% alginate was relatively low (20%), while that released at pH 7.4 increased significantly (86%). In conclusion, the methylcellulose/alginate hydrogel blended with NaCl could be a suitable carrier for site-specific protein drug delivery in the intestine. PMID- 15360307 TI - Synthesis of novel biodegradable cationic polymer: N,N-diethylethylenediamine polyurethane as a gene carrier. AB - A new cationic polymer, N,N-diethylethylenediamine-polyurethane (DEDA-PU), bearing tertiary amines in the backbone and side chains, was synthesized and used as a nonviral vector for gene delivery. The DEDA-PU readily self-assembled with the plasmid DNA (pCMV-betagal) in water and buffer at physiological pH, as determined by agarose gel retardation, dynamic light scattering, zeta potential, atomic force microscopy (AFM), and restriction endonuclease protection assays. The results revealed that DEDA-PU was able to bind with plasmid DNA, yielding positively charged complexes with a size around 100 nm at a DEDA-PU/DNA ratio of 50/1 (w/w). The DEDA-PU/DNA complexes were able to transfect HEK 293 cells in vitro with an efficiency comparable to a well-known gene carrier [poly(2 dimethylaminoethyl methacrylate), PDMAEMA]. The cytotoxicity of DEDA-PU was substantially lower than PDMAEMA. The degradation studies indicated that DEDA-PU degrades hydrolytically in 20 mM HEPES buffer at pH 7.4 with a half-life of approximately 60 h. This study shows that DEDA-PU holds promise as biodegradable polycations for gene delivery and is interesting candidate for further study. PMID- 15360308 TI - Biomacromolecules electrostatic self-assembly on 3-dimensional tissue engineering scaffold. AB - A poly(ethylenimine) (PEI) was employed to obtain a stable positively charged surface on a poly(D,L-lactide) (PDL-LA) tissue engineering scaffold. An extracellular matrix (ECM)-like biomacromolecule, gelatin, was selected as polyelectrolyte and deposit alternately with PEI on the activated PDL-LA scaffold via ESA technique. The zeta-potential result showed alternating charge of polyelectrolytes (PEI/gelatin) layering on PDL-LA microspheres. Quartz crystal microbalance (QCM) measurement further verified the gradual deposition of PEI/gelatin on the PDL-LA thin film. The combination of PEI aminolysis and the layer-by-layer technique was then explored to construct gelatin coating onto the 3-D porous PDL-LA scaffold. Scanning electronic microscopy showed that there is no notable difference between modified and unmodified PLA scaffolds, with regard to the porosity, pore diameter, and scaffold integration. The dual-tunnel confocal laser scanning microscopy indicated uniform gelatin distribution on the inner surface of the 3-D porous scaffold. The gradual build-up of protein layer on scaffold was investigated by radioiodination technique. Chondrocyte was chosen to test the cell behavior on modified and unmodified PDL-LA scaffolds. The results of the cell viability, total intracellular protein content, and cell morphology on the PEI/gelatin multilayers modified PDL-LA scaffold showed to promote chondrocyte growth. Comparing conventional coating methods, polyelectrolyte multilayers are easy and stable to prepare. It may be a promising choice for the surface modification of complex biomedical devices. These very flexible systems allow broad medical applications for drug delivery and tissue engineering. PMID- 15360309 TI - Thermally modulated insulin release from microgel thin films. AB - We describe investigations of thermally triggered insulin release from poly(N isopropylacrylamide-co-acrylic acid) microgel thin films prepared by layer-by layer (LbL) polyelectrolyte assembly. The thermoresponsivity of these films was confirmed using light scattering techniques. Simultaneous monitoring of film collapse and insulin release kinetics shows that deswelling of the films is partially decoupled from macromolecule release and that release is mainly governed by partitioning effects. We hypothesize, however, that film thermoresponsivity plays an important role in that subjection to many thermal cycles enables the embedded peptide to solubilize and subsequently partition through film layers. Direct pulsatile and extended release studies confirm the capability of these films to release bursts of insulin over many cycles, and confirm that the magnitude of the release can be controlled based on film thickness. These insulin-impregnated films are extremely stable with the potential to release constant pulses of peptide for more than 1 month at a time. PMID- 15360310 TI - Rational protein modification leading to resistance of enzymes to TiO2-UV irradiation-induced inactivation. AB - Photoexcited TiO2 degrades biomolecules such as nucleic acids, cell membrane proteins, and enzymes. Stabilization of enzyme activity against the deactivation caused by the combination of TiO2-UV is essential if we are to develop novel hybrid materials exhibiting photocatalytic and biocatalytic activities useful for decontamination applications. In this paper we describe the stabilization of a model enzyme, chymotrypsin, against TiO2-UV-induced deactivation by conjugating the enzyme with UV-absorbing, carboxyl-terminated oligo[2-[3-(2H-benzotriazol-2 yl)-4-hydroxyphenyl]ethyl methacrylate] [oligo(HBMA)-COOH]. Chymotrypsin was completely deactivated within 3 h, whereas the chymotrypsin-oligo(HBMA) conjugate retained > 50% activity even after 5 h of exposure to TiO2-UV (lambdamax 365 nm). The degree of enzyme stabilization induced by the conjugated UV absorber was 2 fold higher than that from the equivalent number of conjugated PEG chains. Spectroscopic characterizations revealed that chymotrypsin-oligo(HBMA) absorbs UV light and initially resists photoexcitation of TiO2. Modified chymotrypsin also exhibited resistance to changes in the secondary structure during the deactivation. This method of stabilizing enzymes against photodegradation could be also useful in photolithographic enzyme immobilizations for sensors and arrays or for stabilization of any UV-sensitive protein. PMID- 15360311 TI - Probing the binding of scutellarin to human serum albumin by circular dichroism, fluorescence spectroscopy, FTIR, and molecular modeling method. AB - The binding of scutellarin with human serum albumin (HSA) was investigated at four temperatures, 296, 303, 310, and 318 K, by fluorescence, circular dichroism (CD), Fourier transform infrared spectroscopy (FT-IR), and molecular modeling study at pH 7.40. The binding parameters were determined by Scatchard's procedure, which are approximately consistent with the results of Stern-Volmer equation. The thermodynamic parameters were calculated according to the dependence of enthalpy change on the temperature as follows: DeltaH degrees is a small negative value (-8.55 kJ/mol), whereas DeltaS degrees is a positive value (65.15 J/mol K). Quenching of the fluorescence HSA in the presence of scutellarin was observed. Data obtained by fluorescence spectroscopy and CD experiment, FT-IR experiment, and molecular modeling method suggested that scutellarin can strongly bind to the HSA and the primary binding site of scutellarin is located in site I of HSA. It is considered that scutellarin binds to site I (subdomain II) mainly by a hydrophobic interaction and there are hydrogen bond interactions between the scutellarin and the residues Arg222 and Arg257. PMID- 15360312 TI - Protein encapsulation via porous CaCO3 microparticles templating. AB - Porous microparticles of calcium carbonate with an average diameter of 4.75 microm were prepared and used for protein encapsulation in polymer-filled microcapsules by means of electrostatic layer-by-layer assembly (ELbL). Loading of macromolecules in porous CaCO3 particles is affected by their molecular weight due to diffusion-limited permeation inside the particles and also by the affinity to the carbonate surface. Adsorption of various proteins and dextran was examined as a function of pH and was found to be dependent both on the charge of the microparticles and macromolecules. The electrostatic effect was shown to govern this interaction. This paper discusses the factors which can influence the adsorption capacity of proteins. A new way of protein encapsulation in polyelectrolyte microcapsules is proposed exploiting the porous, biocompatible, and decomposable microparticles from CaCO3. It consists of protein adsorption in the pores of the microparticles followed by ELbL of oppositely charged polyelectrolytes and further core dissolution. This resulted in formation of polyelectrolyte-filled capsules with protein incorporated in interpenetrating polyelectrolyte network. The properties of CaCO3 microparticles and capsules prepared were characterized by scanning electron microscopy, microelectrophoresis, and confocal laser scanning microscopy. Lactalbumin was encapsulated by means of the proposed technique yielding a content of 0.6 pg protein per microcapsule. Horseradish peroxidase saves 37% of activity after encapsulation. However, the thermostability of the enzyme was improved by encapsulation. The results demonstrate that porous CaCO3 microparticles can be applied as microtemplates for encapsulation of proteins into polyelectrolyte capsules at neutral pH as an optimal medium for a variety of bioactive material, which can also be encapsulated by the proposed method. Microcapsules filled with encapsulated material may find applications in the field of biotechnology, biochemistry, and medicine. PMID- 15360313 TI - pH-sensitive hydrogel based on a novel photocross-linkable copolymer. AB - A pH sensitive hydrogel has been prepared by a UV irradiation technique. Starting polymer was the PHM (poly hydroxyethylaspartamide methacrylated) obtained from polyaspartamide (PHEA) partially derivatized with methacrylic anhydride (MA). This new copolymer has been further derivatized with succinic anhydride (SA) to obtain PHM-SA that has been cross-linked by UV irradiation to form a pH sensitive hydrogel. The network, recovered after washing as a powder, has been been characterized by FT-IR spectrophotometry and particle size distribution analysis. Moreover, to have information about water affinity of the prepared sample, swelling measurements have been carried out in aqueous media mimicking biological fluids. The possibility to employ the prepared hydrogel as a pH-sensitive drug delivery system (DDS) has been investigated. In particular, ibuprofen ((S)(+)4 isobutyl-alpha-methylphenyl-acetic acid), chosen as a model drug, has been entrapped into the PHM-SA hydrogel, and in vitro release studies have showed that its release rate depends on different swelling of the network as a function of the environmental pH. PMID- 15360314 TI - TEMPO-mediated oxidation of native cellulose. The effect of oxidation conditions on chemical and crystal structures of the water-insoluble fractions. AB - Cellulose cotton linter was oxidized with sodium hypochlorite with catalytic amounts of sodium bromide and 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO) under various conditions. After this TEMPO-mediated oxidation, water insoluble fractions were collected and characterized in terms of carboxylate and aldehyde contents, crystallinities and crystal sizes, degrees of polymerization, morphology, and water retention values. Carboxylate and aldehyde groups were introduced into the water-insoluble fractions up to about 0.7 and 0.3 mmol/g, respectively, by the oxidation, where recovery of the water-insoluble fractions were generally higher than 80%. Crystallinities and crystal sizes of cellulose I were nearly unchanged during the oxidation, and thus, carboxylate and aldehyde groups were introduced selectively on crystal surfaces and in disordered regions of the water-insoluble fractions. Water retention values of cotton linter can be increased from 60% to about 280% through the introduction of hydrophilic carboxylate groups and morphological changes from fibrous forms to short fragments by the TEMPO-mediated oxidation. PMID- 15360315 TI - Nanoreinforcement of poly(propylene fumarate)-based networks with surface modified alumoxane nanoparticles for bone tissue engineering. AB - A novel composite material has been fabricated for bone tissue engineering scaffolds utilizing the biodegradable polymer poly(propylene fumarate)/poly(propylene fumarate)-diacrylate (PPF/PPF-DA) and surface-modified carboxylate alumoxane nanoparticles. Various surface-modified nanoparticles were added to the polymer including a surfactant alumoxane, an activated alumoxane, a mixed alumoxane containing both activated and surfactant groups, and a hybrid alumoxane containing both groups within the same substituent. These nanocomposites, as well as polymer resin and unmodified boehmite composites, underwent flexural and compressive mechanical testing and were examined using electron microscopy. Hybrid alumoxane nanoparticles dispersed in PPF/PPF-DA exhibited over a 3-fold increase in flexural modulus at 1 wt % loading compared to polymer resin alone. No significant loss of flexural or compressive strength was observed with increased loading of hybrid alumoxane nanoparticles. These dramatic improvements in flexural properties may be attributed to the fine dispersion of nanoparticles into the polymer and increased covalent interaction between polymer chains and surface modifications of nanoparticles. PMID- 15360316 TI - Synthesis and biologically relevant properties of polyphosphazene polyacids. AB - Polyphosphazene polyacids show potential as immunostimulating compounds and materials for microencapsulation. Their synthesis requires multistep chemical transition from a hydrolytically unstable macromolecular precursor, poly(dichlorophosphazene), to a water-soluble polyelectrolyte. Insufficient synthetic control in these reactions can lead to molecular weight variations and formation of macromolecules with "structural defects" resulting in significant variations in polymer performance. Simple and reproducible "one pot-one solvent" method is reported for the preparation of polyphosphazene polyacids poly[di(carboxylatophenoxy)phosphazene] and its copolymers. Molecular weight characteristics and polymer compositions were studied as a function of reaction parameters. Macromolecular byproducts, incompletely substituted polymers containing hydroxyl groups and partially deprotected polymers containing propyl ester functionalities, were synthesized and characterized. It was demonstrated, that the presence of such groups can affect polymer characteristics, such as hydrolytic degradation profiles, immunostimulating activity, and microsphere forming properties. In vivo studies showed that the immunostimulating activity of polyphosphazene polyacids correlates with the content of acid functionalities in the polymer. PMID- 15360317 TI - Preliminary evaluation of the behavior of fifth-generation thiophosphate dendrimer in biological systems. AB - The quickly growing number of newly synthesized families of dendrimers is not always followed by the information about their properties. Especially, the biological aspect seems not to be explored sufficiently. The aim of this study was to examine the hemotoxicity and cytotoxicity of water-insoluble, fifth generation thiophosphate dendrimer (theoretical Mw = 20,025). We were particularly interested in the impact of dendrimer on human red blood cells structure and membrane integrity. We also evaluated the impact of the thiophosphate-based dendrimer on the growth of nucleated cells represented by Chinese hamster ovary cells (CHO-K1 line). We found that the interaction of dendrimer with erythrocyte membrane proteins triggers echinocytosis but also increases erythrocyte membrane stability and its thermal durability. We observed that the dendrimer moderately stimulated cloning and increased the rate of CHO cellular metabolism. PMID- 15360318 TI - Structural characterization of rubber from jackfruit and euphorbia as a model of natural rubber. AB - A structural study of low molecular weight rubbers from Jackfruit (Artocarpus heterophyllus) and Painted spurge (Euphorbia heterophylla) was carried out as model compounds of natural rubber from Hevea brasiliensis. The rubber content of latex from Jackfruit was 0.4-0.7%, which is very low compared with that of 30-35% in the latex from Hevea tree. The rubber from Jackfruit latex was low molecular weight with narrow unimodal molecular weight distribution (MWD), whereas that obtained from E. heterophylla showed very broad MWD. The 1H and 13C NMR analyses showed that Jackfruit rubber consists of a dimethylallyl group and two trans isoprene units connected to a long sequence of cis-isoprene units. The alpha terminal group of Jackfruit rubber was presumed to be composed of a phosphate group based on the presence of 1H NMR signal at 4.08 ppm corresponding to the terminal =CH-CH2OP group. PMID- 15360319 TI - Transport and tensile properties of compression-molded wheat gluten films. AB - Mechanical and transport properties were assessed on wheat gluten films with a glycerol content of 25-40%, prepared by compression molding for 5-15 min at temperatures between 90 and 130 degrees C. Effects of storing the films up to 24 days, in 0 and 50% relative humidity (RH), were assessed by tensile measurements. The films were analyzed with respect to methanol zero-concentration diffusivity, oxygen permeability (OP), water vapor permeability (WVP), Cobb60 and sodium dodecyl sulfate (SDS) solubility coupled with sonication. The SDS solubility and methanol diffusivity were lower at the higher molding temperature. Higher glycerol content resulted in higher OP (90-95% RH), WVP, and Cobb60 values, due to the plasticizing and hygroscopic effects. Higher glycerol contents gave a lower fracture stress, lower Young's modulus, lower fracture strain, and less strain hardening. The mold time had less effect on the mechanical properties than mold temperature and glycerol content. The fracture stress and Young's modulus increased and the fracture strain decreased with decreasing moisture content. PMID- 15360320 TI - Immobilization of rennet from Mucor miehei via its sugar chain. Its use in milk coagulation. AB - A successful strategy for the immobilization of rennet from Mucor miehei has been developed. The strategy is based on the immobilization of the enzyme, via their sugar chains at high ionic strength on aminated supports having primary amino groups with a very low pK value. The rennet was covalently immobilized via sugar chains (previously oxidized with periodate), which act as natural spacer arms and allow a very high percentage of rennet activity to be kept against small (H-Leu Ser-p-nitro-Phe-Nle-Ala-Leu-OMe.TFA (98%)) and macromolecular substrates (k casein) (78%). The use of tailor-made aminated support was critical to obtain good stability values, because using fully aminated supports achieved much lower thermostability values than using 50% aminated supports. The optimized derivative was utilized to hydrolyze casein in milk. To prevent the coagulation of the milk in the presence of the derivative, the reaction was performed at 4 degrees C (where hydrolyzed casein did not precipitate). Then the hydrolyzed milk was filtered and latter on heated to 30 degrees C, achieving a similar aggregate to the one achieved with soluble rennet. PMID- 15360321 TI - Poly(vinyl chloride)-coated sol-gels for studying the effects of nitric oxide release on bacterial adhesion. AB - Nitric oxide (NO) releasing sol-gel materials coated with poly(vinyl chloride) (PVC) films exhibit increased stability at ambient and physiological temperatures. The polymer overcoat, however, reduces the NO fluxes by 5-35% over the initial week of release. The variation in NO fluxes between unmodified and PVC-coated sol-gels is negligible after 7 days. The PVC polymeric layer provides controlled surface chemistry for systematic studies of the effects of NO release on bacterial adhesion. As an example, the adhesion of Pseudomonas aeruginosa and Proteus mirabilis at PVC-coated NO-releasing sol-gels is investigated. A direct NO dependence on the reduction of P. aeruginosa adhesion is observed for NO fluxes up to 20 pmol cm(-2) s(-1). Although decreased by 50% in the presence of NO release, P. mirabilis adhesion does not appear to correlate to the flux of NO release. PVC-coated NO-releasing sol-gels may prove useful for studying the effects of localized NO release on other biological and chemical systems. PMID- 15360322 TI - Study of the synthesis, crystallization, and morphology of poly(ethylene glycol) poly(epsilon-caprolactone) diblock copolymers. AB - Poly(ethylene glycol)-poly(epsilon-caprolactone) diblock copolymers PEG-PCL were synthesized by ring-opening polymerization of epsilon-caprolactone using monomethoxy poly(ethylene glycol) as the macroinitiator and calcium ammoniate as the catalyst. Obvious mutual influence between PEG and PCL crystallization was studied by altering the relative block length. Fixing the length of the PEG block (Mn = 5000) and increasing the length of the PCL block, the crystallization temperature of the PCL block rose gradually from 1 to about 35 degrees C while that of the PEG block dropped from 36 to -6.6 degrees C. Meanwhile, the melting temperature of the PCL block went up from 30 to 60 degrees C, while that of the PEG block declined from 60 to 41 degrees C. If the PCL block was longer than the PEG block, the former would crystallize first when cooling from a molten state and led to obviously imperfect crystallization of PEG and vice versa. And they both crystallized at the same temperature, if their weight fractions were equal. We found that the PEG block could still crystallize at -6.6 degrees C even when its weight fraction is only 14%. A unique morphology of concentric spherulites was observed for PEG5000-PCL5000. According to their morphology and real-time growth rates, it is concluded that the central and outer sections in the concentric spherulites were PCL and PEG, respectively, and during the formation of the concentric spherulite, the PEG crystallized quickly from the free space of the PCL crystal at the earlier stage, followed by outgrowing from the PCL spherulites in the direction of right angles to the circle boundaries until the entire area was occupied. PMID- 15360323 TI - Solution properties of chitin in alkali. AB - The solution properties of alpha-chitin dissolved in 2.77 M NaOH are discussed. Chitin samples in the weight-average molecular weight range 0.1 x 10(6) g/mol to 1.2 x 10(6) g/mol were prepared by heterogeneous acid hydrolysis of chitin. Dilute solution properties were measured by viscometry and light scattering. From dynamic light scattering data, relative similar size distributions of the chitin samples were obtained, except for the most degraded sample, which contained aggregates. Second virial coefficients in the range 1 to 2 x 10(-3) mL.mol.g(-2) indicated that 2.77 M NaOH is a good solvent to chitin. The Mark-Houwink-Sakurada equation and the relationship between the z-average radius of gyration (Rg) and the weight-average molecular weight (Mw) were determined to be [eta] = 0.10Mw0.68 (mL.g(-1)) and Rg = 0.17Mw0.46 (nm), respectively, suggesting a random-coil structure for the chitin molecules in alkali conditions. These random-coil structures have Kuhn lengths in the range 23-26 nm. PMID- 15360324 TI - Combined biotin-terpyridine systems: a new versatile bridge between biology, polymer science and metallo-supramolecular chemistry. AB - Biotin, a well-known binding unit for the proteins avidin and streptavidin, was combined with the chelating ligand terpyridine via polymeric and nonpolymeric spacers. An omega-amino-functionalized terpyridyl-poly(ethylene glycol) was prepared and utilized for complex formation with iron(II), nickel(II), and ruthenium(II) ions. The biocompatibility of the complex formation was investigated in aqueous media. Moreover, biotin was functionalized with a methoxy poly(ethylene glycol) as a model system. The compounds were characterized by UV/vis and NMR spectroscopy as well as MALDI-TOF mass spectrometry. The systems represent a new combination of strong noncovalent binding units from both biology and synthetic supramolecular chemistry. PMID- 15360325 TI - Shear characteristics, miscibility, and topography of sodium caseinate monoglyceride mixed films at the air-water interface. AB - In this contribution, we are concerned with the study of structure, topography, and surface rheological characteristics (under shear conditions) of mixed sodium caseinate and monoglycerides (monopalmitin and monoolein) at the air/water interface. Combined surface chemistry (surface film balance and surface shear rheometry) and microscopy (Brewster angle microscopy, BAM) techniques have been applied in this study to mixtures of insoluble lipids and sodium caseinate spread at the air-water interface. At a macroscopic level, sodium caseinate and monoglycerides form an heterogeneous and practically immiscible monolayer at the air-water interface. The images from BAM show segregated protein and monoglyceride domains that have different topography. At surface pressures higher than that for the sodium caseinate collapse, this protein is displaced from the interface by monoglycerides. These results and those derived from interfacial shear rheology (at a macroscopic level) appear to support the idea that immiscibility and heterogeneity of these emulsifiers at the interface have important repercussions on the shear characteristics of the mixed films, with the alternating flow of segregated monoglyceride domains (of low surface shear viscosity, etas) and protein domains (of high etas) across the canal. PMID- 15360326 TI - Molecular motions in chitosan studied by dielectric relaxation spectroscopy. AB - Neutralized and nonneutralized chitosan films subject to different thermal treatments were studied by dielectric relaxation spectroscopy from -130 to +150 degrees C in the frequency range between 20 Hz and 1 MHz. Two main relaxation processes, both arrhenian type, were detected: process I at temperatures below 0 degrees C with a mean activation energy of 49 +/- 1 kJ mol(-1), which has the characteristics of a secondary relaxation process related with local chain dynamics, and process II observable at higher temperatures with an activation energy of 94 +/- 2 kJ mol(-1), correlated with dc conductivity, which is found in dried polysaccharides systems. Process I is always observed in neutralized chitosan, but it is strongly depleted in the wet nonneutralized form. Although the location of process I is independent of NH2/NH3+ side group, process II deviates to higher temperatures with dryness in both chitosan forms, being located at lower temperatures in nonneutralized chitosan. PMID- 15360763 TI - Knowledge management in pediatric pain: mapping on-line expert discussions to medical literature. AB - Clinical decision-making can be vastly improved with the availability of the right medical knowledge at the right time. This concept paper presents a knowledge management re-search program to (a) identify, capture and organize the tacit knowledge inherent within on-line problem-solving discussions between pediatric pain practitioners; (b) establish linkages between topic-specific pediatric pain discussions and corresponding published medical literature on children's pain available at PubMed--i.e. linking tacit expert knowledge to explicit medical literature; and (c) make these knowledge re-sources available to pediatric pain practitioners via the WWW for timely access to various modalities of clinical knowledge. PMID- 15360764 TI - Data mart based research in heart surgery: challenges and benefit. AB - For many new medical research questions in heart surgery comprehensive and large data bases are essential. We discuss typical challenges for the integration of real-time and legacy data stored in multiple unconnected hospital information systems (HIS). Furthermore the HIS are often operated by autonomous departments whose data base structures are subject to occasional modifications. We present a solution which integrates and consolidates all research relevant data in a data mart without imposing any considerable operational or maintenance contract liability risk for the existing HIS. The problems of partial consistency and partial redundancy in the data are discussed. The data mart system serves multiple purposes: beside clinical reporting and quality assessment, the preparation steps for comprehensive studies are enormously simplified. PMID- 15360765 TI - Using a Bayesian network to predict the probability and type of breast cancer represented by microcalcifications on mammography. AB - Since the widespread adoption of mammographic screening in the 1980's there has been a significant increase in the detection and biopsy of both benign and malignant microcalcifications. Though current practice standards recommend that the positive predictive value (PPV) of breast biopsy should be in the range of 25 40%, there exists significant variability in practice. Microcalcifications, if malignant, can represent either a non-invasive or an invasive form of breast cancer. The distinction is critical because distinct surgical therapies are indicated. Unfortunately, this information is not always available at the time of surgery due to limited sampling at image-guided biopsy. For these reasons we conducted an experiment to determine whether a previously created Bayesian network for mammography could predict the significance of microcalcifications. In this experiment we aim to test whether the system is able to perform two related tasks in this domain: 1) to predict the likelihood that microcalcifications are malignant and 2) to predict the likelihood that a malignancy is invasive to help guide the choice of appropriate surgical therapy. PMID- 15360766 TI - Patterns of usage for a Web-based clinical information system. AB - Understanding how clinicians are using clinical information systems to assist with their everyday tasks is valuable to the system design and development process. Developers of such systems are interested in monitoring usage in order to make enhancements. System log files are rich resources for gaining knowledge about how the system is being used. We have analyzed the log files of our Web based clinical information system (WebCIS) to obtain various usage statistics including which WebCIS features are frequently being used. We have also identified usage patterns, which convey how the user is traversing the system. We present our method and these results as well as describe how the results can be used to customize menus, shortcut lists, and patient reports in WebCIS and similar systems. PMID- 15360767 TI - Aligning words in French-English non-parallel medical texts: effect of term frequency distributions. AB - In this paper, we present a method for aligning words based on a statistical model of word distribution similarity. The basis underlying our method is that there is a correlation between the patterns of word co-occurrences in texts of different languages. Using automatically downloaded pages from different medical web sites and a combined bilingual lexicon of general and medical terms as language sources, a similarity score is assigned to each proposed translated pair of words, based on the distributional contexts of these two words. We vary several parameters of the method. Experimental results confirm a positive effect of frequency, show that medical words are better handled than less specialized words, and do not evidence a clear influence of context window size. Future directions for improvement include working with very large, part-of-speech tagged corpora. PMID- 15360768 TI - A guideline management system. AB - This paper describes the architecture of NewGuide, a guide-line management system for handling the whole life cycle of a computerized clinical practice guideline. NewGuide components are organized in a distributed architecture: an editor to formalize guidelines, a repository to store them, an inference engine to implement guidelines instances in a multi-user environment, and a reporting system storing the guidelines logs in order to be able to completely trace any individual physician guideline-based decision process. There is a system "central level" that maintains official versions of the guidelines, and local Healthcare Organizations may download and implement them according to their needs. The architecture has been implemented using the Java 2 Enterprise Edition (J2EE) platform. Simple Object Access Protocol (SOAP) and a set of con-tracts are the key factors for the integration of NewGuide with healthcare legacy systems. They allow maintaining unchanged legacy user interfaces and connecting the system with what-ever electronic patient record. The system functionality will be illustrated in three different contexts: homecare-based pressure ulcer prevention, acute ischemic stroke treatment and heart failure management by general practitioners. PMID- 15360769 TI - NCI Thesaurus: using science-based terminology to integrate cancer research results. AB - Cancer researchers need to be able to organize and report their results in a way that others can find, build upon, and relate to the specific clinical conditions of individual patients. NCI Thesaurus is a description logic terminology based on current science that helps individuals and software applications connect and organize the results of cancer research, e.g., by disease and underlying biology. Currently containing some 34,000 concepts--covering chemicals, drugs and other therapies, diseases, genes and gene products, anatomy, organisms, animal models, techniques, biologic processes, and administrative categories--NCI Thesaurus serves applications and the Web from a terminology server. As a scalable, formal terminology, the deployed Thesaurus, and associated applications and interfaces, are a model for some of the standards required for the NHII (National Health Information Infrastructure) and the Semantic Web. PMID- 15360770 TI - Synthesis of elementary single-disease recommendations to support guideline-based therapeutic decision for complex polypathological patients. AB - Situations managed by clinical practice guidelines (CPGs) usually correspond to general descriptions of theoretical patients that suffer from only one disease in addition to the specific pathology CPGs focus on. The lack of decision support for complex multiple-disease patients is usually transferred to computer-based systems. Starting from the GEM-encoded instance of CPGs, we developed a module that automatically generated IF-THEN-WITH decision rules. A two-stage unification process has been implemented. All the rules whose IF-part is in partial matching with a patient clinical profile were triggered. A synthesis of triggered rules has then been performed to eliminate redundancies and incoherences. All remaining, eventually contradictory, recommendations were displayed to physicians leaving them the responsibility of handling the controversy and thus the opportunity to control the therapeutic decision. PMID- 15360771 TI - Real world performance of approximate string comparators for use in patient matching. AB - Medical record linkage is becoming increasingly important as clinical data is distributed across independent sources. To improve linkage accuracy we studied different name comparison methods that establish agreement or disagreement between corresponding names. In addition to exact raw name matching and exact phonetic name matching, we tested three approximate string comparators. The approximate comparators included the modified Jaro-Winkler method, the longest common substring, and the Levenshtein edit distance. We also calculated the combined root-mean square of all three. We tested each name comparison method using a deterministic record linkage algorithm. Results were consistent across both hospitals. At a threshold comparator score of 0.8, the Jaro-Winkler comparator achieved the highest linkage sensitivities of 97.4% and 97.7%. The combined root-mean square method achieved sensitivities higher than the Levenshtein edit distance or long-est common substring while sustaining high linkage specificity. Approximate string comparators increase deterministic linkage sensitivity by up to 10% compared to exact match comparisons and represent an accurate method of linking to vital statistics data. PMID- 15360772 TI - A neural network model of lexicalization for simulating the anomic naming errors of dementia patients. AB - Word-finding difficulty (anomia) is the most common linguistic deficit in dementia. It is often measured by picture naming tasks as naming a picture taps all the major processes in word production, i.e., activation of a concept, retrieval of lexical-semantic information on that concept, retrieval of the corresponding word form and articulation. Naming and naming errors have extensively been simulated by neural network models of lexicalization (see e.g. [1,2]). A common feature of these models is that they are static, i.e. non learning. However, naming is a dynamic process that changes as a function of normal learning or re-learning after neural damage. These important patterns cannot be caught by the static models of lexicalization. Therefore we have developed a learning model of lexicalization based on multi-layer-perceptron (MLP) neural networks. We tested the model by fitting it to the naming data of 22 Finnish-speaking dementia patients and 19 neurologically intact control subjects. The tests showed an excellent fit between the model's and the subjects naming response distributions. Thus our model seems be suitable to simulate naming disorders of dementia patients. PMID- 15360773 TI - The NLM Gateway: a metasearch engine for disparate resources. AB - The U.S. National Library of Medicine (NLM) has created a metasearch engine called the NLM Gateway at the URL "gateway.nlm.nih.gov". The Gateway allows the user to issue one search that takes place on multiple NLM retrieval engines. A composite result set is presented in several categories of information: journal citations; books, serials and audiovisuals; consumer health; meeting abstracts; and other collections. PMID- 15360774 TI - Facilitating knowledge maintenance of clinical guidelines and protocols. AB - Clinical protocols and guidelines are widely used in the medi-cal domain to improve disease management techniques. Different software systems are in development to support the de-sign and the execution of such guidelines. The bottleneck in the guideline software developing process is the transformation of the text-based clinical guidelines into a formal representation, which can be used by the execution software. This paper introduces a method and a tool that was designed to provide a solution for that bottleneck. The so-called Guideline Markup Tool (GMT) facilitates the translation of guidelines into a formal representation written in XML. This tool enables the protocol designer to create links between the original guideline and its formal representation and ease the editing of guidelines applying design patterns in the form of macros. The usefulness of our approach is illustrated using GMT to edit Asbru protocols. We performed a usability study with eight participants to examine the usefulness of the GMT and of the Asbru macros, which showed that the proposed approach is very appropriate to author and maintain clinical guidelines. PMID- 15360775 TI - Knowledge sharing and information integration in healthcare using ontologies and deductive databases. AB - This paper describes a method for using Semantic Web technologies for sharing knowledge in healthcare. It combines deductive databases and ontologies, so that it is possible to extract knowledge that has not been explicitly declared within the database. A representation of the UMLS (Unified Medical Language System) Semantic Network and Metathesaurus was created using the RDF standard, in order to represent the basic medical ontology. The inference over the knowledge base is done by the TRI-DEDALO System, a deductive data-base created to query and update RDF based knowledge sources as well as conventional relational databases. Finally, an ontology was created for the Brazilian National Health Card data interchange format, a standard to capture and transmit health encounter information throughout the country. This paper demonstrates how this approach can be used to integrate heterogeneous information and to answer complex queries in a real world environment. PMID- 15360776 TI - The MeSH translation maintenance system: structure, interface design, and implementation. AB - The National Library of Medicine (NLM) produces annual editions of the Medical Subject Headings (MeSH). Translations of MeSH are often done to make the vocabulary useful for non-English users. However, MeSH translators have encountered difficulties with entry vocabulary as they maintain and update their translation. Tracking MeSH changes and updating their translations in a reasonable time frame is cumbersome. NLM has developed and implemented a concept centered vocabulary maintenance system for MeSH. This system has been extended to create an interlingual database of translations, the MeSH Translation Maintenance System (MTMS). This database allows continual updating of the translations, as well as facilitating tracking of the changes within MeSH from one year to another. The MTMS interface uses a Web-based design with multiple colors and fonts to indicate concepts needing translation or review. Concepts for which there is no exact English equivalent can be added. The system software encourages compliance with the Unicode standard in order to ensure that character sets with native alphabets and full orthography are used consistently. PMID- 15360777 TI - Decision support for patient care: implementing cybernetics. AB - The application of principles and methods of cybernetics permits clinicians and managers to use feedback about care effectiveness and resource expenditure to improve quality and to control costs. Keys to the process are the specification of therapeutic goals and the creation of an organizational culture that supports the use of feedback to improve care. Daily feedback on the achievement of each patient's therapeutic goals provides tactical decision support, enabling clinicians to adjust care as needed. Monthly or quarterly feedback on aggregated goal achievement for all patients on a clinical pathway provides strategic decision support, enabling clinicians and managers to identify problems with supposed "best practices" and to test hypotheses about solutions. Work is underway at Vanderbilt University Medical Center to implement feedback loops in care and management processes and to evaluate the effects. PMID- 15360778 TI - Modeling and analyzing biomedical processes using workflow/Petri Net models and tools. AB - Computer simulation enables system developers to execute a model of an actual or theoretical system on a computer and analyze the execution output. We have been exploring the use of Petri Net (PN) tools to study the behavior of systems that are represented using three kinds of biomedical models: a biological workflow model used to represent biological processes, and two different computer interpretable models of health care processes that are derived from clinical guidelines. We developed and implemented software that maps the three models into a single underlying process model (workflow), which is then converted into PNs in formats that are readable by several PN simulation and analysis tools. These analysis tools enabled us to simulate and study the behavior of two biomedical systems: a Malaria parasite invading a host cell, and patients undergoing management of chronic cough. PMID- 15360779 TI - Models and inference methods for clinical systems: a principled approach. AB - Previous papers have argued for the existence of three different models in many clinical information systems--for the medical record, for inference in guidelines, and for concepts and re-usable facts. This paper presents a principled approach to deciding which information belongs in each model based on the nature of the queries or inference to be performed: necessary or contingent, open or closed world, algorithmic vs heuristic. It then discusses an important class of systems--"ontologically indexed knowledge bases"--and issues of metadata within this framework. PMID- 15360780 TI - A knowledge-based time-oriented active database approach for intelligent abstraction, querying and continuous monitoring of clinical data. AB - Query and interpretation of time-oriented medical data involves two subtasks: Temporal-reasoning--intelligent analysis of time-oriented data, and temporal maintenance--effective storage, query, and retrieval of these data. Integration of these tasks into one system, known as temporal-mediator, has been proven to be beneficial to biomedical applications such as monitoring, therapy, quality assessment, visualization and exploration of time-oriented data. One potential problem in existing temporal-mediation approaches is lack of sufficient responsiveness when querying or continuously monitoring the database for complex abstract concepts that are derived from the raw data, especially regarding a large patient group. We propose a new approach: the knowledge-based time-oriented active database, a temporal extension of the active-database concept, and a merger of temporal reasoning and temporal maintenance within a persistent database framework. The approach preserves the efficiency of databases in handling data storage and retrieval, while enabling specification and performance of complex temporal reasoning using an incremental-computation approach. We implemented our approach within the Momentum system. Initial experiments are encouraging; an evaluation is underway PMID- 15360781 TI - Medical expert systems developed in j.MD, a Java based expert system shell: application in clinical laboratories. AB - Growing complexity of diagnostic tests, combined with increased workload, stringent laboratory accreditation demands, continuous shortening of turn-around time and budget restrictions have forced laboratories to automate most of their iterative tasks. Introduction of artificial intelligence by means of expert systems has gained an important place in this automation process. Different parts of clinical laboratory activity can benefit from their implementation and the present project deals with one aspect, namely the clinical interpretation of diagnostic tests. This paper describes how j.MD, a new Java based expert system shell, was used to reprogram the expert system for interpretation of amylase isoenzyme patterns that has been in use for many years in our laboratory, and that was originally programmed in Pro.MD, a Prolog based expert system shell. One of the most important advantages of the j.MD system is its bidirectional link with the laboratory information system. This project shows how expert systems for the interpretation of complex diagnostic tests that demand specific expertise can become an integrated part of the automated clinical chemistry lab. PMID- 15360782 TI - Contextual annotation of web pages for interactive browsing. AB - With the information on the World Wide Web and in specialized databases exploding, researchers and physicians are in dire need to browse efficiently though the large corpus of information resources in their field of interest. The focus is not any longer to find everything related to your interest, but it shifts to zooming in, based on context and expanding again in neighboring knowledge domains. This paper describes an attempt to develop a completely new, interactive way of browsing distributed corpora of information without the need for multiple different queries in different information resources. Classical search engines generally treat search requests in isolation. The results for a given query are identical, and do not automatically take on board the context in which the user made the request. The system described here explores implicit contexts as obtained from the document that the user is reading. The new approach merges the searching and browsing into one combined "read-and-search" mode and alleviates the shift users are normally forced to between searching and reading. PMID- 15360783 TI - Using knowledge management practices to develop a state-of-the-art digital library. AB - Diffusing knowledge management practices within an organization encourages and facilitates reuse of the institution's knowledge commodity. Following knowledge management practices, the Eskind Biomedical Library (EBL) has created a Digital Library that uses a holistic approach for integration of information and skills to best represent both explicit and tacit knowledge inherent in libraries. EBL's Digital Library exemplifies a clear attempt to organize institutional knowledge in the field of librarianship, in an effort to positively impact clinical, research, and educational processes in the medical center. PMID- 15360784 TI - Cost-effectiveness analysis of colorectal cancer screening strategies in Singapore: a dynamic decision analytic approach. AB - A dynamic decision analytic framework using local statistics and expert's opinions is put to study the cost-effectiveness of colorectal cancer screening strategies in Singapore. It is demonstrated that any of the screening strategies, if implemented, would increase the life expectancy of the population of 50 to 70 years old. The model also determined the normal life expectancy of this population to be 76.32 years. Overall, Guaiac Fecal Occult Blood Test (FOBT) is most cost effective at SGD162.11 per life year saved per person. Our approach allowed us to model problem parameters that change over time and study the utility measures like cost and life expectancy for specific age within the range of 50- 69 through to 70 years old. PMID- 15360786 TI - Computerization of guidelines: a knowledge specification method to convert text to detailed decision tree for electronic implementation. AB - The initial step for the computerization of guidelines is the knowledge specification from the prose text of guidelines. We describe a method of knowledge specification based on a structured and systematic analysis of text allowing detailed specification of a decision tree. We use decision tables to validate the decision algorithm and decision trees to specify and represent this algorithm, along with elementary messages of recommendation. Edition tools are also necessary to facilitate the process of validation and workflow between expert physicians who will validate the specified knowledge and computer scientist who will encode the specified knowledge in a guide-line model. Applied to eleven different guidelines issued by an official agency, the method allows a quick and valid computerization and integration in a larger decision support system called EsPeR (Personalized Estimate of Risks). The quality of the text guidelines is however still to be developed further. The method used for computerization could help to define a framework usable at the initial step of guideline development in order to produce guidelines ready for electronic implementation. PMID- 15360785 TI - Adherence to computerized clinical reminders in a large healthcare delivery network. AB - OBJECTIVE: To evaluate clinicians' adherence with clinical reminders (CRs) across multiple ambulatory practice settings in an integrated health care network. MATERIALS AND METHODS: Adherence rate to 15 CRs, integrated into a computerized patient record system, was evaluated for 451 clinicians in 49 clinics from eight Veterans Affairs (VA) medical centers. RESULTS: Overall, mean rate of adherence to CRs for all clinics was 86.2%, with a range of 66.59% to 97.08% (P<.001). The hepatitis C risk assessment reminder was found to have the highest overall adherence rate (95.9%) and the tobacco use cessation had the lowest adherence rate (62.9%). Mean adherence rate for all reminders was 80.34% (P<.001). Mean adherence rate for individual clinicians was 82.6%, with a range of 29% to 100%. CONCLUSION: While overall adherence to CRs was high, there is significant variation by clinic, individual clinician and individual CR. Understanding this variation is critical in directing future efforts to improve the contribution of computerized CRs to quality and cost-effectiveness of care, and to decrease undesirable variation in clinical practice. Further research is needed to systematically evaluate clinician, reminder and systems related factors that influence adherence to CRs. PMID- 15360787 TI - Restoring the patient therapeutic history from prescription data to enable computerized guideline-based decision support in primary care. AB - The application of a guideline-based therapeutic strategy in the context of a chronic disease requires a clear picture, at the appropriate level of abstraction, of a patient's particular therapeutic history. However, like most clinical data, information on past treatments is often incompletely specified. We propose temporal abstraction mechanisms based on a simple and heuristic treatment of temporal indeterminacy of period bounds. Applied to prescription data, continuity and simultaneousness of treatments have to be characterized. Even if limited, our method restores the therapeutic history of a patient, with episodes of mono-, bi-, and tritherapies in order to adequately position her within the guideline therapeutic strategy. PMID- 15360788 TI - Evaluating provider adherence in a trial of a guideline-based decision support system for hypertension. AB - Measurement of provider adherence to a guideline-based decision support system (DSS) presents a number of important challenges. Establishing a causal relationship between the DSS and change in concordance requires consideration of both the primary intention of the guideline and different ways providers attempt to satisfy the guideline. During our work with a guideline-based decision support system for hypertension, ATHENA DSS, we document a number of subtle deviations from the strict hypertension guideline recommendations that ultimately demonstrate provider adherence. We believe that understanding these complexities is crucial to any valid evaluation of provider adherence. We also describe the development of an advisory evaluation engine that automates the interpretation of clinician adherence with the DSS on multiple levels, facilitating the high volume of complex data analysis that is created in a clinical trial of a guideline-based DSS. PMID- 15360789 TI - Convergence on a standard for representing clinical guidelines: work in health level seven. AB - Concern regarding patient safety and practice variation has focused attention on clinical guidelines as a way of influencing the behavior of health care providers in order to improve patient outcomes. Despite the abundant number of guidelines produced, their use has been limited, in part because the necessary knowledge is relatively inaccessible at the point of care. In turn, this has led researchers to make guidelines computable so that they can be employed in information systems that provide tailored decision support. A number of different efforts have been mounted to create a standard formalism. Health Level Seven (HL7) is a key international standards development organization. Ongoing work in HL7 attempts to synthesize the best aspects of these efforts, producing along the way shareable components of a guideline representation, such as a common expression language and standard data model. These shareable components require in turn an information architecture that makes available a range of patient data encoded in an appropriate format. Convergence on a standard guideline formalism will facilitate use of guidelines and thus enhance patient care. PMID- 15360790 TI - Effectiveness of an electronic medical record clinical quality alert prepared by off-line data analysis. AB - We tested whether off-line data analysis, instead of event monitoring, was a viable method for initiating a clinical quality alert. A cohort of patients eligible for an alert was identified by off-line data analysis and a flag was set in their ambulatory Electronic Medical Records. One hundred clinicians were randomly assigned either to a control group or to a group that received the alert when viewing the electronic medical record of eligible patients. Primarily due to actions of their clinicians, 315 of the 580 patients (54.3%) seen by alerted clinicians were no longer eligible for the alert at the end of the one month study, compared to 128 of the 496 patients (25.8%) seen by control clinicians (p<.001). When not alerted, Allied Health clinicians were less likely than physicians to prescribe aspirin, but they responded similarly to the alert. There were no differences in response by specialty or gender of the clinician. Off-line data analysis proved to be an effective method of initiating a clinical alert. PMID- 15360791 TI - Vaidurya--a concept-based, context-sensitive search engine for clinical guidelines. AB - A major problem in the effective use of clinical guidelines is fast and accurate access at the point of care. Thus, we are developing a digital electronic guideline library (DeGeL) and a set of tools for incremental conversion of free text guide-lines into increasingly machine-comprehensible representations, which support automated application. Even if guidelines are represented in electronic fashion, care providers need to be able to quickly retrieve the guidelines that best fit the clinical situation at hand. We describe Vaidurya, a search and retrieval engine that exploits the hybrid nature of guideline representation in the DeGeL architecture. Vaidurya can use not only free-text keywords, but also multiple semantic indices along which the guidelines are classified, and the mark up of guidelines in DeGeL, using the semantic roles of one or more guideline representation languages. Preliminary evaluation of Vaidurya in a standard information task and a large guide-line repository is encouraging; formal evaluation is under way. PMID- 15360792 TI - Detailed clinical models for sharable, executable guidelines. AB - The goal of shareable, executable clinical guidelines is both worthwhile and challenging. One of the largest hurdles is that of representing the necessary clinical information in a precise and shareable manner. Standard terminologies and common information models, such as the HL7 RIM, are necessary, they are not sufficient. In addition, common detailed clinical models are needed to give precise semantics and to make the task of mapping between models manageable. We discuss the experience of the SAGE project related to detailed clinical models. PMID- 15360793 TI - A methodology for modular representation of guidelines. AB - Computer-based clinical practice guidelines often need to be modified when medical knowledge evolves or when guidelines are implemented in a local setting with specific constraints and preferences. To enable easy modifications to guidelines and maintain their integrity, we have developed a methodology for modular representation of guidelines. Using this approach, we create guidelines in a hierarchical and modular manner. We use the Axiomatic Design methodology to facilitate the development of independent modules. Design matrices capture the interactions among modules. The design matrices can be used during guideline modification to create a change process and to enable identification of other modules that are affected by a change to a module. We implemented this modular knowledge representation approach by incorporating it into the Guideline Interchange Format (GLIF) language. We applied this approach to encode parts of three outdated guidelines released during 2000-2001, and we revised these designs to model updated releases of the guideline. Qualitative and quantitative metrics were developed to assess the types of changes made to the encoded guidelines. PMID- 15360794 TI - Development of computerized guidelines for the management of chronic diseases allowing to position any patient within recommended therapeutic strategies. AB - Chronic diseases are complex to manage. One reason comes from the difficulty to synchronize a patient's therapeutic history with the guideline-based sequence of treatments. We propose to represent guideline knowledge as a two-level decision tree, a clinical level describing theoretical clinical situations and a therapeutic level formalizing the different steps of corresponding recommended therapeutic strategies. Guideline-based strategies are first represented as bidimensional matrices structured in lines of therapy and levels of therapeutic intention. A revised version introducing levels of therapeutic combination is then developed. The therapeutic level is operationalized for any patient therapeutic history to provide the next best step of treatment. Evaluated on actual patient records, our system proved to impact physicians' decisions in 78% of the cases and led to a significant improvement of their compliance with recommendations. PMID- 15360795 TI - Toward improved guideline quality: using the COGS statement with GEM. AB - The Conference on Guideline Standardization (COGS) was convened to create a standardized documentation checklist for clinical practice guidelines in an effort to promote guideline quality and facilitate implementation. The statement was created by a multidisciplinary panel using a rigorous consensus development methodology. The Guideline Elements Model (GEM) provides a standardized approach to representing guideline documents using XML. In this work, we demonstrate the sufficiency of GEM for describing COGS components. Using the mapping between COGS and GEM elements we built an XSLT application to examine a guideline's adherence (or non-adherence) to the COGS checklist. Once a guideline has been marked up according to the GEM hierarchy, its knowledge content can be reused in multiple ways. PMID- 15360796 TI - Description and status update on GELLO: a proposed standardized object-oriented expression language for clinical decision support. AB - A major obstacle to sharing computable clinical knowledge is the lack of a common language for specifying expressions and criteria. Such a language could be used to specify decision criteria, formulae, and constraints on data and action. Al though the Arden Syntax addresses this problem for clinical rules, its generalization to HL7's object-oriented data model is limited. The GELLO Expression language is an object-oriented language used for expressing logical conditions and computations in the GLIF3 (GuideLine Interchange Format, v. 3) guideline modeling language. It has been further developed under the auspices of the HL7 Clinical Decision Support Technical Committee, as a proposed HL7 standard., GELLO is based on the Object Constraint Language (OCL), because it is vendor-independent, object-oriented, and side-effect-free. GELLO expects an object-oriented data model. Although choice of model is arbitrary, standardization is facilitated by ensuring that the data model is compatible with the HL7 Reference Information Model (RIM). PMID- 15360797 TI - A context-adaptable approach to clinical guidelines. AB - One of the most relevant obstacles to the use and dissemination of clinical guidelines is the gap between the generality of guidelines (as defined, e.g., by physicians' committees) and the peculiarities of the specific context of application. In particular, general guidelines do not take into account the fact that the tools needed for laboratory and instrumental investigations might be unavailable at a given hospital. Moreover, computer-based guideline managers must also be integrated with the Hospital Information System (HIS), and usually different DBMS are adopted by different hospitals. The GLARE (Guideline Acquisition, Representation and Execution) system addresses these issues by providing a facility for automatic resource-based adaptation of guidelines to the specific context of application, and by providing a modular architecture in which only limited and well-localised changes are needed to integrate the system with the HIS at hand. PMID- 15360798 TI - Modeling guidelines for integration into clinical workflow. AB - The success of clinical decision-support systems requires that they are seamlessly integrated into clinical workflow. In the SAGE project, which aims to create the technological infra-structure for implementing computable clinical practice guide-lines in enterprise settings, we created a deployment-driven methodology for developing guideline knowledge bases. It involves (1) identification of usage scenarios of guideline-based care in clinical workflow, (2) distillation and disambiguation of guideline knowledge relevant to these usage scenarios, (3) formalization of data elements and vocabulary used in the guideline, and (4) encoding of usage scenarios and guideline knowledge using an executable guideline model. This methodology makes explicit the points in the care process where guideline-based decision aids are appropriate and the roles of clinicians for whom the guideline-based assistance is intended. We have evaluated the methodology by simulating the deployment of an immunization guideline in a real clinical information system and by reconstructing the workflow context of a deployed decision-support system for guideline-based care. We discuss the implication of deployment-driven guideline encoding for sharability of executable guidelines. PMID- 15360799 TI - Analysis of Dutch general practice guidelines for inconsistencies with respect to the management of cardiovascular disease risk factors. AB - Presenting guidelines relating to cardiovascular disease (CVD) in a clinical decision support system (CDSS) is viewed as a mechanism for introducing evidence based practice in to daily practice. Analysis of guidelines on consistency is needed before constructing such a system. We analyzed 74 electronically available Dutch general practice guidelines for statement inconsistencies with respect to the management of CVD, and for each inconsistency, whether incomplete cross referencing existed between the guidelines. RESULTS: Six of the 74 guidelines had either CVD or CVD risk factors as subject of the guideline, all adhering to a common structure corresponding to four sections reflecting distinctive clinical management stages, which correspond to the subjective, objective, assessment and plan stages of SOAP methodology for structured patient data recording. Ten statement inconsistencies were found and for each a reference inconsistency was present. CONCLUSIONS: In a CDSS, statement inconsistencies will lead to poor system functioning. With electronically published guidelines, guideline developers could link statements in guidelines, to comparable statements in previously issued guidelines, facilitating the review process with respect to inconsistencies in related guidelines, and consistent cross referencing. Furthermore, it could enable the updating of existing guidelines with new evidence. Despite the determined statement inconsistencies, the authors would be able to construct a CDSS based on the DCGP guidelines dealing with the management of CVD risk factors PMID- 15360800 TI - Child Health Improvement through Computer Automation: the CHICA system. AB - Clinical guidelines are prevalent but frequently not used. Computer reminder systems can improve adherence to guidelines but have not been widely adopted. We present a computer-based decision support system that combines these elements: 1) pediatric preventive care guidelines encoded in Arden Syntax; 2) a dynamic, scannable paper user interface; and 3) a HL7-compliant interface to existing electronic medical record systems. The result is a system that both delivers "just in time" patient-relevant guidelines to physicians during the clinical encounter and accurately captures structured data from all who interact with the system. The system performs these tasks while remaining sensitive to the workflow constraints of a busy outpatient pediatric practice. PMID- 15360801 TI - Community-wide information systems design: concepts and an illustration. AB - Models for community-wide information systems design help insure the development of a health information infrastructure that supports life-long, community centric, patient focused care. Collaborating with community health planning groups provides a reasonable starting point for such endeavors. We review existing approaches to community-based information systems design, draw design principles from the experiences of past innovations, and describe our team's experience in partnering with a community health planning group to establish a community-wide design approach to information systems design. Key themes include (1) partnerships; (2) brokering and integrating existing initiatives; and (3) creating business models for sustainability PMID- 15360802 TI - A quantitative and qualitative evaluation of LISA, a decision support system for chemotherapy dosing in childhood acute lymphoblastic leukaemia. AB - OBJECTIVES: To assess the acceptability to clinicians of a web-based decision support system designed to assist with dosage adjustments during maintenance therapy for childhood Acute Lymphoblastic Leukaemia (ALL), and to evaluate the potential impact of the system on decision-making and dosage calculations. DESIGN: Balanced-block crossover experiment with simulated cases; questionnaire study and semi-structured interviews. PARTICIPANTS: 36 clinicians with differing experience in the management of ALL. INTERVENTIONS: Subjects were asked to decide on appropriate levels of chemotherapy dosing for 8 simulated cases, 4 using the LISA decision support system, 4 using conventional paper-based records and guidance. MAIN OUTCOME MEASURES: Number of protocol-consistent dosage decisions made; time taken to manage each case; accuracy of dosage calculations; subjects' opinions as to whether or not they would use the system in practice. ADDITIONAL OUTCOME MEASURES: Functions subjects would like to see in an idealised system; subjects' satisfaction with the implementation of the functions provided by LISA; qualitative data on issues subjects felt would impact upon the successful deployment of the system. PMID- 15360803 TI - Implementation and initial evaluation of a Web-based nurse order entry system for multidrug-resistant tuberculosis patients in Peru. AB - Socios En Salud uses directly observed therapy to treat a majority of the multidrug-resistant tuberculosis in Peru. The nurses play an important role in this community-based model as the patients' primary care givers. Since nurses, rather than physicians, are involved in patients' daily care, we developed a nurse-order entry system to test whether such a system would improve the accuracy and quality of medication data. We compared regimen information from patient electronic medical records, paper charts and pharmacy records. After a two-month training period on the new system, we conducted the trial for 52 days in two of Lima's six geographic treatment areas, and re-reviewed the three sources of medication data. We measured the error rates after the trial period and found there was no significant difference in the control group's (Lima Este), error rate (8.6% vs. 6.9%, P=0.66) after the trial. The intervention group (Lima Callao), however, showed a significant drop in the error rate (17.4% vs. 3.1%, P=0.0074) after the same time interval. Additionally, the nurse expressed satisfaction with the order entry system and its ease of use. The decrease in error rates and user satisfaction regarding the system are promising measures of our order entry system's success. PMID- 15360804 TI - EyePACS: an open source clinical communication system for eye care. AB - PURPOSE: EyePACS is an application for communicating and archiving eye-related patient information, images, and diagnostic data. We studied how users adopted the system in diverse clinical settings. METHODS: 53 clinicians and 142 students uploaded cases over 2.5 years from 6 pilot sites: a university teaching clinic, a university glaucoma clinic, an urban private optometric practice, a rural elderly care facility, a diabetic management program, and an eye hospital in India. RESULTS: EyePACS collected 1,122 cases. Users employed it for informal "curbside" consults in 17% of cases. Other uses of the system were: 1) to replace telephone and fax referrals to a retinal specialist (10%), 2) as part of ocular teleconsultations and diabetic retinopathy screening (31%), 3) for education via digital grand rounds and evaluation of students (32%), and 4) for research (10%). CONCLUSION: EyePACS has been used successfully for consults and education in diverse settings. The resulting database of digital cases serves as a searchable reference for clinicians. PMID- 15360805 TI - System-wide surveillance for clinical encounters by patients previously identified with MRSA and VRE. AB - Methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) have emerged as major infection control problems worldwide. Patients previously infected or colonized with MRSA or VRE need to be identified and often isolated as soon as they visit a health care facility. Infection control personnel usually are not aware when these patients enter their facilities. We developed a system-wide surveillance system to alert infection control personnel when patients with previous MRSA or VRE cultures from LDS Hospital have subsequent clinical encounters at any inpatient or outpatient facility at Intermountain Health Care (IHC). This paper describes this system and includes the results from an initial study on the potential epidemiological benefits provided to help improve patient care. The study found that patients with previous MRSA and VRE had subsequent encounters at 62 different IHC facilities up to 304 miles away from 1 day to over 5 years later. In addition, the new surveillance system was able to alert infection control personnel when ever these patients visited any IHC inpatient or outpatient facility. PMID- 15360806 TI - Improving cancer related symptom management with collaborative healthware. AB - We conducted a feasibility study of an innovative Internet-based system, designed to support collaboration between patients and providers around cancer related symptom management. Our objective was to demonstrate the system's potential to enhance communication and clinical decision making between patients and their providers. Considering the high prevalence of cancer related symptoms and their impact on patients' quality of life, enhanced collaboration regarding assessment and treatment could significantly reduce illness burden. Twenty-seven patients with known malignancy and twenty-nine oncology clinicians were interviewed to identify functional requirements for the system. Patients reported the ability, willingness and desire to use a computer to help them manage their cancer related symptoms. Physicians recognized the potential of this system to improve communication and collaboration around symptom management, but voiced concern regarding a potential increased workload. Nurse providers had greater interest in reviewing symptoms online than did their physician counterparts. Patients in this study believe that using an Internet based system to report their cancer related symptoms would improve the quality of the care they receive. Our findings suggest that both patients and clinicians are concerned about the management of cancer related symptoms and would use Internet-based tools if they were shown to improve care. PMID- 15360807 TI - TOSCA-Imaging--developing Internet based image processing software for screening and diagnosis of diabetic retinopathy. AB - The primary aim of TOSCA-Imaging, which was a part of the TOSCA Project financed by EU's Fifth Framework IST Programme, was to develop Internet based software and image data bases for screening and diagnosis of diabetic retinopathy and implementing it into a real life situation. The work consisted of: 1) Construction of an Internet based communication platform for transmitting and analyzing retinal images. 2) Implementation of routines for detecting the first microaneurysm (transition from normal to pathologic), detecting patients needing referral for treatment (presence of venous beading or hard exudates near the fovea), and for serial analysis (image alignment). 3) Construction of a reference image data base. A preliminary validation showed that decisions that depended on a precise detection of individual lesions, e.g. the detection of normality, had a sensitivity and specificity of around 80%, whereas decisions that depended on the detection of lesion patterns, e.g. clinically significant macular oedema, had a sensitivity and specificity of more than 95%. Validation of the reference image data base by double grading by two expert graders suggested a sensitivity and a specificity of just below 90% for any lesion and of more than 95% for predicting the overall retinopathy grade. TOSCA-Imaging has succeeded in its primary aim of developing Internet based software and implementing it into a real life situation, integrating work within image processing done in four different European countries (England, Germany, Ireland, and Denmark) to be accessed from one Internet web site. PMID- 15360808 TI - An evaluation of automatically generated briefings of patient status. AB - We report on an evaluation of MAGIC, a system that automatically generates briefings of patient status after coronary bypass surgery, completed in the Cardio Thoracic Intensive Care Unit at New York Presbyterian Hospital. Through enhancements in system design, robustness and speed, we compared information obtained by nurses against two briefings, one automatically generated by MAGIC and one provided by physicians upon the patient's arrival to the ICU. Our results show that MAGIC and the physician briefing provide a substantial increase in the amount of information than is available prior to the patient's arrival and that the information MAGIC provides is accurate. In many aspects, MAGIC out-performs the physician briefing; information is reported earlier and is always available. We conclude that MAGIC provides the CT ICU staff early on with a better assessment of the patient's status than in current practice and allows them to better prepare for the patient's arrival. PMID- 15360809 TI - Facilitated lipid management using interactive e-mail: preliminary results of a randomized controlled trial. AB - We describe the design and implementation of a clinical decision support system for ambulatory hyperlipidemia management. Despite readily available and evidence based guidelines, cholesterol control remains persistently sub-optimal in clinical practice. "FastTrack" is an integrated, multi-faceted informatics tool to reduce barriers to effective cholesterol management by proactively integrating pertinent clinical information, evidence-based decision support and a simple means to act via a single FastTrack e-mail. We tested the intervention in a randomized, controlled trial involving 14 physicians and 276 patients. In an interim analysis, we found that physicians on average were able to review and act upon 9 patient e-mails in less than 15 minutes. Significantly more intervention patients were initiated on a statin therapy or had their dose increased (15.3% vs. 2.3%, p < 0.001). Impact of the intervention on LDL levels will be assessed at trial completion. PMID- 15360810 TI - Validation of a Web-based prognostic system for breast cancer. AB - A website has been published which allows the user to enter information on prognostic factors for a patient with breast cancer, and instantly obtain a survival curve based on outcome data of prior cases with a matching prognostic factor profile. The source for the survival data is a Finnish nationwide series (the FinProg series) of 2,842 women diagnosed with breast cancer in 1991-2. The purpose of this study was to compare survival estimates based on the FinProg database, with estimates for breast cancer patients from the US, obtained from the SEER public-use database and the same time-period. Results show that a reasonable level of agreement between estimates can be reached, by the use of large, unselected databases, and that significantly different estimates were obtained in only 2 of 19 analyzed prognostic profiles. The current system could be used to share important knowledge on outcome between researchers and clinicians at different institutions, and be used in the decision-making process concerning treatment of patients with breast cancer. PMID- 15360811 TI - A knowledge management system for new drug submission by pharma-industries. AB - The pharma-industries are facing a number of crucial business issues to improve operational excellence in product time-to-market and wide regulatory compliance. These organizations own, produce, and manipulate a lot of knowledge. The new regulations by Health Authorities (HA) to pharma-industries should make the content and format of new drug application uniform worldwide. In this paper we suggest a novel approach of a pharma-industry to capture, process, and transmit clinical data electronically. The approach begins with an analysis of the knowledge generation points, some of them being outside the company. Implementations are grounded on the use of a de facto standard platform being Microsoft, having acceptable cost levels. The proposed infrastructure is integrated into existing company environment and technological platform, minimizing cost and risks, but improving efficiency and efficacy of new drug dossier compilation. PMID- 15360812 TI - e-clinic: an electronic triage system for the management of type 2 Diabetes Mellitus. AB - Diabetes Mellitus represents a heterogeneous group of metabolic disorders and its complications are associated with increased morbidity and early mortality. The explosive increase of the number of patients with diabetes, and several recommendations from milestone studies such as the United Kingdom Prospective Study (UKPDS), the Diabetes Prevention Program (DPP) and the Finnish Prevention Study place further constraint on an already overstretched service. This paper examines the care flow process in the current diabetes management and explores the potential role of new technologies and methodologies currently available to support the highly complex clinical requirements. The paper proposes a concept of e-clinic as an electronic triage system with integrated healthcare record, embedded with a decision support system to analyse patients' data and provide a continuous monitoring system according to agreed clinical guidelines. PMID- 15360813 TI - Executing clinical practice guidelines using the SAGE execution engine. AB - We report the first successful test of an interoperable guideline execution engine that interprets encoded clinical guideline content and executes that content via functions of a target clinical information system (CIS). For this test, an exemplar immunization guideline was encoded in the SAGE guideline model using standards-based information models and terminologies. This guideline content was subsequently executed using the prototype SAGE guideline execution engine, which interacts through standards-based VMR/Action services to instantiate real-time guideline recommendations via existing functions of the target CIS. In this paper, we describe our test implementation and highlight the significance and implications of each component of our deployment architecture PMID- 15360814 TI - A database for Chinese outpatients with rheumatic diseases. AB - Although several databases have been developed in rheumatology aimed at profiling the morbidity pattern of rheumatic diseases, or to capture the detailed clinical and outcome information of patients with a specific rheumatic disease, there is no database in China, as yet, for capturing visit-related health information of all outpatients with rheumatic diseases. In this project, a relational database was developed for the rheumatic outpatient clinic of Peking Union Medical College Hospital (PUMCH). The primary goal of this database is to collect and organize visit-related information of outpatients with rheumatic diseases to meet information needs of clinical research, health care continuity and teaching of rheumatic diseases. The implementation of this database can improve health services, medical records management and clinical research of outpatients with rheumatic disease at PUMCH. PMID- 15360815 TI - Learning Boolean queries for article quality filtering. AB - Prior research has shown that Support Vector Machine models have the ability to identify high quality content-specific articles in the domain of internal medicine. These models, though powerful, cannot be used in Boolean search engines nor can the content of the models be verified via human inspection. In this paper, we use decision trees combined with several feature selection methods to generate Boolean query filters for the same domain and task. The resulting trees are generated automatically and exhibit high performance. The trees are understandable, manageable, and able to be validated by humans. The subsequent Boolean queries are sensible and can be readily used as filters by Boolean search engines. PMID- 15360816 TI - The NLM Indexing Initiative's Medical Text Indexer. AB - The Medical Text Indexer (MTI) is a program for producing MeSH indexing recommendations. It is the major product of NLM's Indexing Initiative and has been used in both semi-automated and fully automated indexing environments at the Library since mid 2002. We report here on an experiment conducted with MEDLINE indexers to evaluate MTI's performance and to generate ideas for its improvement as a tool for user-assisted indexing. We also discuss some filtering techniques developed to improve MTI's accuracy for use primarily in automatically producing the indexing for several abstracts collections. PMID- 15360817 TI - Overcoming information overload: an information system for the primary care physician. AB - Primary Care Office InSite (PCOI) is a Web-based intranet application that provides ready access to a collection of information useful in primary care. The PCOI Web site was developed by, and is widely used within, the Massachusetts General Hospital (MGH) and its affiliated community practices. Over 1600 users logged 60,000 separate sessions in the past year. The site contains clinical practice guidelines, patient educational material, drug prescription and cost information and referral information, all designed for use during routine patient care activity. A continuing and dynamic collaboration between the users and developers of the system, just-in-time access and a "one-stop portal" to a variety of resources have resulted in a very high approval rating by the users. This paper discusses the important characteristics of the Web site, specific functions that improve the workflow of the practicing clinician, and limitations of the present implementation. PMID- 15360818 TI - Practical considerations for exploiting the World Wide Web to create infobuttons. AB - BACKGROUND: We are studying ways to provide automated, context-specific links (called "infobuttons") between clinical information systems (CIS) and other information resources available on the World Wide Web. As part of this work, we observed the information needs that arose when clinicians used a CIS and we classified those needs into generic questions. We then sought general methods for accessing information resources to answer the questions. METHODS: For each generic question, we identified a satisfactory resource and then developed a method for retrieving from it the information relevant to the question. We then studied these methods to characterize them into general approaches. RESULTS: We identified six general approaches and describe them in detail. These approaches range in complexity from simple, hard-coded links to intelligent agents and calculators. CONCLUSION: Web-based information resources can be exploited using a relatively small number of methods, although the specific methods require custom solutions. Standard methods for accessing Web-based resources would simplify the task of linking to CISs. PMID- 15360819 TI - Context-sensitive medical information retrieval. AB - Substantial medical data such as pathology reports, operative reports, discharge summaries, and radiology reports are stored in textual form. Databases containing free-text medical narratives often need to be searched to find relevant information for clinical and research purposes. Terms that appear in these documents tend to appear in different contexts. The con-text of negation, a negative finding, is of special importance, since many of the most frequently described findings are those denied by the patient or subsequently "ruled out." Hence, when searching free-text narratives for patients with a certain medical condition, if negation is not taken into account, many of the retrieved documents will be irrelevant. The purpose of this work is to develop a methodology for automated learning of negative context patterns in medical narratives and test the effect of context identification on the performance of medical information retrieval. The algorithm presented significantly improves the performance of information retrieval done on medical narratives. The precision im-proves from about 60%, when using context-insensitive retrieval, to nearly 100%. The impact on recall is only minor. In addition, context-sensitive queries enable the user to search for terms in ways not otherwise available PMID- 15360820 TI - Application of a Medical Text Indexer to an online dermatology atlas. AB - Clinical dermatology cases are presented as images and semi-structured text describing skin lesions and their relationships to disease. Metadata assignment to such cases is hampered by lack of a standardized dermatology vocabulary and facilitated methods for indexing legacy collections. In this pilot study descriptive clinical text from Dermatlas, a Web-based repository of dermatology cases, was indexed to Medical Subject Heading (MeSH) terms using the National Library of Medicine's Medical Text Indexer (MTI). The MTI is an automated text processing system that derives ranked lists of MeSH terms to describe the content of medical journal citations using knowledge from the Unified Medical Language System (UMLS) and from MEDLINE. For a representative, random sample of 50 Dermatlas cases, the MTI frequently derived MeSH indexing terms that matched expert-assigned terms for Diagnoses (88%), Lesion Types (72%), and Patient Characteristics (Gender and Age Groups, 62% and 84% respectively). This pilot demonstrates the potential for extending the MTI to automate indexing of clinical case presentations and for using MeSH to describe aspects of clinical dermatology. PMID- 15360821 TI - Text mining functional keywords associated with genes. AB - Modern experimental techniques provide the ability to gather vast amounts of biological data in a single experiment (e.g. DNA microarray experiment), making it extremely difficult for the researcher to interpret the data and form conclusions about the functions of the genes. Current approaches provide useful information that organizes or relates genes, but a major shortcoming is they either do not address specific functions of the genes or are constrained by functions predefined in other databases, which can be biased, incomplete, or out of-date. We extended Andrade and Valencia's method [1] to statistically mine functional keywords associated with genes from MEDLINE abstracts. The MEDLINE abstracts are analyzed statistically to score and rank keywords for each gene using a background set of words for baseline frequencies. We generally got very good functional keyword information about the genes we tested, which was confirmed by searching for the individual keywords in context. The keywords extracted by our algorithm reveal a wealth of potential functional concepts, which were not represented in existing public databases. We feel that this approach is general enough to apply to medical and biological literature to find other relationships: drugs vs. genes, risk-factors vs. genes, etc. PMID- 15360822 TI - Clinicians' assessments of the usefulness of online evidence to answer clinical questions. AB - Family physicians' experience of using an online evidence retrieval system in routine clinical work was measured. Participants' assessments regarding the usefulness of the online evidence to answer their clinical questions were determined using three feedback channels: voluntary online feedback during the trial; randomized prompted feedback during a specific online evidence search; and responses in a post-trial survey. Using an analysis of the web logs, search outcome was related to patterns of searching in terms of number of searches and documents viewed. The majority of clinicians' searches were related to patient care and information retrieved was rated as useful in answering these questions. On average clinicians conducted two searches and viewed one document per clinical question. Search sessions which resulted in "completely answering" a clinical question involved significantly fewer searches than less successful searches. The three feedback channels provided different information but consistent overall results. The findings indicate that clinicians' assessments of online evidence use are linked to search patterns and online feedback is useful for measuring the effectiveness of systems. PMID- 15360823 TI - Evolution of medical informatics in bibliographic databases. AB - Medical informatics became a medical specialty during the last years and this is evidenced by a great amount of journal articles regarding the subject published worldwide. We compared the presentation of Medical Informatics in two different bibliographic databases: MEDLINE and LILACS (Latin American and Caribbean Literature on the Health Sciences). Previous studies described how Medical Informatics was represented in MEDLINE, but we wanted to compare it to a regional database as LILACS. We search both databases completely (MEDLINE 1966 -2002 and LILACS 1982-2002) using the keyword "Medical Informatics" as MeSH term in MEDLINE and as DeCS term in LILACS, and we added "medical informatics" as text word and analyzed the references obtained as results. We found that MEDLINE properly represents the impact of Medical Informatics in non-Latin-American international journals, but lacks of a considerable amount of articles from this region, while LILACS, although in comparison it is smaller in size, has more articles regarding the subject. So we think that LILACS properly represents the specialty in Latin America and the Caribbean Region. PMID- 15360824 TI - Scenario-based assessment of physicians' information needs. AB - Physicians' information needs routinely arise during their practice. Several studies have demonstrated that a useful way to understand the nature of the needs is to examine questions posed by physicians during the course of medical care. This paper presents an analysis of clinical questions collected when physicians were engaged in reviewing clinical cases. The main objective of the study was to investigate characteristics of the physicians' information needs to provide insight into the development of a conceptual guidance approach in information retrieval. The analysis focused on categories, search contexts, and patterns of the questions with respect to the scenarios. The results of the study showed that physicians' information needs exhibited distinct characteristics according to the scenarios, and that their needs could be expressed with a relatively small number of question patterns. These observations supported our approach of using patterns of information needs in facilitating digital information access. PMID- 15360825 TI - Developing optimal search strategies for detecting clinically relevant qualitative studies in MEDLINE. AB - BACKGROUND: The growing interest in qualitative research within the evidence based practice framework highlights the need for accurate search strategies to enhance the retrieval of qualitative studies. To date, little work has been done on developing optimal search filters for retrieving qualitative studies. The current study extends our earlier work, on developing optimal search strategies, to include qualitative studies. OBJECTIVE: To develop optimal search strategies for detecting clinically relevant qualitative studies in MEDLINE in the publishing year 2000. DESIGN: Comparison of the retrieval performance of methodologic search strategies in MEDLINE with a manual review ("gold standard") of each article for each issue of 161 core health care journals for the year 2000. METHODS: 6 experienced research assistants who had been trained and intensively calibrated reviewed all issues of 161 journals for the publishing year 2000. Each article was systematically classified for "format" (whether it was an original study, review article, general article, or case report), "interest" (whether or not it was of interest to the health care of humans), and "purpose" (whether it pertained to therapy, diagnosis, prognosis, causation, economics, costs, or clinical prediction; was of a qualitative nature; or was about something else). Search strategies were developed for all purpose categories, including qualitative studies. MAIN OUTCOME MEASURES: The sensitivity (recall), specificity, precision, and accuracy of single and combinations of search terms. RESULTS: 49,028 articles were identified after matching the hand search records with the data downloaded from MEDLINE, of which 366 (0.75%) were classified as qualitative. Combinations of search terms reached peak sensitivities of 95%. Compared with the best single term, a three-term strategy increased sensitivity for qualitative studies by 23.6% (absolute increase), but with some loss of specificity when sensitivity was maximized. When search terms were combined to optimize sensitivity and specificity, both these values peaked above 90%. CONCLUSION: Several search strategies can achieve high performance in retrieving qualitative studies from MEDLINE. PMID- 15360826 TI - The specification of a frame-based medical terminological system in Protege. AB - A medical terminological system (TS) is essentially a representation of concepts, attributes and relationships pertaining to medical terms. Although the requirements and structures of TSs have enjoyed some attention in the literature, the actual implementation of TSs consisted so far of ad-hoc approaches starting from scratch. Recently, Protege has been suggested as a software environment for the development of knowledge-based systems. In Protege various component types interact, and knowledge can be represented at various levels. This paper investigates how to specify a frame-based TS in Protege and demonstrates this in a specific application in the intensive care. Our approach is characterized by the utilization of a conceptual framework for understanding TSs and mapping its components onto Protege constructs. This results in specifications of knowledge components for the implementation of terminological systems. The significance of the work stems from the generality of the specifications, thus facilitating the principled design and development of terminological systems in various medical domains PMID- 15360827 TI - Coping with the variability of medical terms. AB - OBJECTIVES: To cope with medical terms, which present a high variability of expression through a single natural language, in the sense that any term may be reformulated in hundred of different ways. METHODS: A typology of term variants is presented as a systematic approach in order to favour the implementation of an exhaustive solution. Then, an algorithm able to handle all variants is designed. RESULTS: Using MetaMap, single terms are analyzed with a success rate varying between 68 and 88 %; the algorithm presented in this paper improves this situation. CONCLUSIONS: This experience shows that a semantic driven method, based on a thesaurus, provides a satisfactory solution to the problem of variability of a single term. The presented typology is representative of most variants in a language. PMID- 15360828 TI - Aligning knowledge sources in the UMLS: methods, quantitative results, and applications. AB - The UMLS Semantic Network and Metathesaurus are two complementary knowledge sources. While many studies compare relationships across the two structures, their alignment has never been attempted. We applied two methods based on lexical and conceptual similarity to aligning the Semantic Network with the UMLS Metathesaurus. Approximately two thirds of the semantic types could be aligned by lexical similarity. Conceptual similarity suggested mappings in all but ten cases. Potential applications enabled by the alignment are discussed, namely auditing the consistency between the Semantic Network and the Metathesaurus and extending the Semantic Network downwards. The relative contribution and limitations of the two methods used for the alignment are also discussed. PMID- 15360829 TI - Embedding clinical indicators into nursing documentation. AB - The Methodist Healthcare System of San Antonio audits completeness and accuracy of nursing assessments. Between 1996 and 2001, regardless of software enhancements and education, completeness of risk assessments hovered at 80% or less. Accuracy of risk scores were in question due to paste functionality. In review, it became apparent that many risk assessment indicators were already an intrinsic part of nursing systems assessment. This project embedded weighted indicators invisibly within systems assessment. Risk scores then automatically calculate and display. This approach decreased documentation queries and increased accuracy of risk assessments. Results were validated using concurrent manual review. Skin assessment demonstrated a 96% accuracy rate with 100% completeness of documentation. Fall assessment resulted in a 2.5% miss rate and 100% completeness of record. 100% of high risk patients identified had appropriate care plan problems. Recommendations are to further explore embedded indicators in software design. The study demonstrated a) decrease in nursing documentation queries b) increase in completeness of record c) increase in accuracy of record and d) increase in accuracy of care plan. PMID- 15360830 TI - Evolution of a Foundational Model of Physiology: symbolic representation for functional bioinformatics. AB - We describe the need for a Foundational Model of Physiology (FMP) as a reference ontology for "functional bioinformatics". The FMP is intended to support symbolic lookup, logical inference and mathematical analysis by integrating descriptive, qualitative and quantitative functional knowledge. The FMP will serve as a symbolic representation of biological functions initially pertaining to human physiology and ultimately extensible to other species. We describe the evolving architecture of the FMP, which is based on the ontological principles of the BioD biological description language and the Foundational Model of Anatomy (FMA). PMID- 15360831 TI - A relation-centric query engine for the Foundational Model of Anatomy. AB - The Foundational Model of Anatomy (FMA), a detailed representation of the structural organization of the human body, was constructed to support the development of software applications requiring knowledge of anatomy. The FMA's focus on the structural relationships between anatomical entities distinguishes it from other current anatomical knowledge sources. We developed Emily, a query engine for the FMA, to enable users to explore the richness and depth of these relationships. Preliminary analysis suggests that Emily is capable of correctly processing real world anatomical queries provided they have been translated into a constrained form suitable for processing by the query engine. PMID- 15360832 TI - Kaiser Permanente's Convergent Medical Terminology. AB - This paper describes Kaiser Permanente's (KP) enterprise-wide medical terminology solution, referred to as our Convergent Medical Terminology (CMT). Initially developed to serve the needs of a regional electronic health record, CMT has evolved into a core KP asset, serving as the common terminology across all applications. CMT serves as the definitive source of concept definitions for the organization, provides a consistent structure and access method to all codes used by the organization, and is KP's language of interoperability, with cross mappings to regional ancillary systems and administrative billing codes. The core of CMT is comprised of SNOMED CT, laboratory LOINC, and First DataBank drug terminology. These are integrated into a single poly-hierarchically structured knowledge base. Cross map sets provide bi-directional translations between CMT and ancillary applications and administrative billing codes. Context sets provide subsets of CMT for use in specific contexts. Our experience with CMT has lead us to conclude that a successful terminology solution requires that: (1) usability considerations are an organizational priority; (2) "interface" terminology is differentiated from "reference" terminology; (3) it be easy for clinicians to find the concepts they need; (4) the immediate value of coded data be apparent to clinician user; (5) there be a well defined approach to terminology extensions. Over the past several years, there has been substantial progress made in the domain coverage and standardization of medical terminology. KP has learned to exploit that terminology in ways that are clinician-acceptable and that provide powerful options for data analysis and reporting. PMID- 15360833 TI - On parts and holes: the spatial structure of the human body. AB - Spatial representation and reasoning is a central component of medical informatics. The spatial concepts most often used in medicine are not the quantitative, point-based concepts of classical geometry, but rather qualitative relations among extended objects such as body parts. A mereotopology is a formal theory of qualitative spatial relations, such as parthood and connection. This paper considers how an extension of mereotopology which includes also location relations can be used to represent and reason about the spatial structure of the human body. PMID- 15360834 TI - WRAPIN: new generation health search engine using UMLS knowledge sources for MeSH term extraction from health documentation. AB - To realize the potential of the Internet as a source of valuable healthcare information, for the general public, patients or practitioners, it is imperative to establish a validation system based on standards of quality. The WRAPIN project (World-wide online Reliable Advice to Patients and Individuals) from the European Community has this ambitious goal. WRAPIN is a federating system for medical information with an editorial policy of intelligently sharing quality and professional information. The WRAPIN project has two main axes: the efficient and intelligent search of information and the assertion of the trustworthiness of content. This article presents the scientific challenges involved in extracting the knowledge from text-based information in order to better manage the knowledge and the rest of the retrieval proc-ess. Our innovative approach is to efficiently extract MeSH terms from the analyzed documents exploiting UMLS knowledge sources. A benefit has been measured when comparing extraction results. Even if the evaluation is made with a limited corpus, this research work proposes heuristics that can be validated to the whole biomedical domain, and possibly enhanced by the adjunction of other methods. PMID- 15360835 TI - Improving information prescription to parents of premature infants through an OWL based knowledge mediator. AB - In the Baby CareLink system, information prescription plays an important role in preparing parents of premature infants for the eventual discharge to home of their children. However, the prescription process requires scarce clinician time in order to dispense information, and can become cumbersome as content bases grow in size. We describe the development of an OWL-based knowledge mediator to facilitate information prescription. We describe 1) the initial development of a clinical vocabulary for neonatology using OWL-DL; 2) the reuse of the vocabulary to represent prototypical premature infants and their typical clinical problems and treatments; 3) the software components used to integrate terminology and inferencing services with Baby CareLink. We demonstrate multifaceted uses for description logics in a clinical application, reuse of a base vocabulary for domain knowledge representation, and use of the OWL language in representing clinical vocabularies. We believe that semi-automated knowledge mediation will enhance the process of electronic information prescription using large clinical content collections. PMID- 15360836 TI - Model once, use multiple times: reusing HL7 domain models from one domain to the other. AB - The Dutch national ICT institute in healthcare (www.nictiz.nl) developed a domain information model for perinatology, to map to the Health Level 7 Reference Information Model (HL7 RIM). This model was constructed with the intention to make it reusable for other domains in order to achieve a national infrastructure based on the HL7 RIM. In two projects, the perinatology model was reused, which lead to some changes to better express the domain. It proves valuable to have a kind of generic domain model, which can be adapted easily to other domains. In these projects, information for hart infarction (cardiology) and stroke (neurology) was modelled. Few domain specific changes where necessary, which could be made following the same methodology. Both the models and the methodology prove to be re-usable. PMID- 15360837 TI - Achieving "source transparency" in the UMLS Metathesaurus. AB - The UMLS Metathesaurus is a syntactically uniform, concept-based, semantically enhanced representation of many of the world's authoritative biomedical vocabularies. Released several times a year, the Metathesaurus is becoming a common, longitudinally maintained source of the current versions of these vocabularies. As vocabularies become standards for reimbursement, reporting, interoperation, and use by applications, the vocabulary obtained from the Metathesaurus must be consistent with that obtainable from each vocabulary's authority. Effective with the first 2004 release, the Metathesaurus represents new and updated sources "transparently"--both users and applications are able to "see" each vocabulary in the Metathesaurus without any of the small losses of information introduced by abstractions used in previous versions. Thus, the Metathesaurus can continue to provide its many semantic and lexical value-added features while guaranteeing that original sources will be "visible" in intact form. Vocabulary users and application developers will benefit from the enhancements and economies of scale offered by the Metathesaurus, while preserving distinctions between content provided by external authorities and content added as part of the Metathesaurus development and maintenance process. PMID- 15360838 TI - Nursing in the information age: status quo and future of ICT use in German hospitals. AB - Hospital information systems (HIS) should give support to nurses in their clinical and managerial duties. Though there are statistical data on the current use of HIS systems we know only little about the numbers of nursing modules implemented. We therefore conducted a nationwide survey in Germany (n = 2182) on the current state and future plans of HIS modules including nursing applications (response rate of 27.6 %). The findings show that management applications (84 % accounting) are still more frequent than clinical applications, in particular clinical patient record systems (19 %). What applied for HIS modules in general held also true for nursing on a lower level. Whereas 51 % of the hospitals had rostering systems in place only 6 % used care planning software. Priorities and plans for the future reveal no change in the rank order of systems. We argue that in order for clinical documentation and planning systems to catch up they must be immediately rewarding for the clinicians in their daily need for information PMID- 15360839 TI - Using symbolic knowledge in the UMLS to disambiguate words in small datasets with a naive Bayes classifier. AB - Current approaches to word sense disambiguation use and combine various machine learning techniques. Most refer to characteristics of the ambiguous word and surrounding words and are based on hundreds of examples. Unfortunately, developing large training sets is time-consuming. We investigate the use of symbolic knowledge to augment machine-learning techniques for small datasets. UMLS semantic types assigned to concepts found in the sentence and relationships between these semantic types form the knowledge base. A naive Bayes classifier was trained for 15 words with 100 examples for each. The most frequent sense of a word served as the baseline. The effect of increasingly accurate symbolic knowledge was evaluated in eight experimental conditions. Performance was measured by accuracy based on 10-fold cross-validation. The best condition used only the semantic types of the words in the sentence. Accuracy was then on average 10% higher than the baseline; however, it varied from 8% deterioration to 29% improvement. In a follow-up evaluation, we noted a trend that the best disambiguation was found for words that were the least troublesome to the human evaluators. PMID- 15360840 TI - Evaluating an ICNP Web-based nursing documentation system. AB - The purpose of this paper is to present the evaluation of a web based nursing documentation system which helps nursing professionals and students to study ICNP Greek beta 2 version and to create nursing diagnoses, interventions and outcomes by selecting relevant terms from ICNP. This advanced web-based ICNP browser enables the authentication of the user, and the creation of a type of nursing care plan through the use of ICNP. It provides different ways for creating nursing diagnoses, interventions and outcomes, by selecting relevant ICNP terms through the tree-structure view, index and search components of the tool. The evaluation of this nursing documentation system is carried out by a survey among nursing professionals working in nursing units in hospitals. The results which have derived from this survey are presented. Moreover, conclusions regarding the future changes that should be made in the system are reported. PMID- 15360841 TI - U.S. Department of Veterans Affairs Enterprise Reference Terminology strategic overview. AB - The Veterans Health Affairs (VHA) branch of the Department of Veterans Affairs has undertaken an Enterprise Reference Terminology (ERT). VHA, arguably the largest integrated healthcare provider in the United States, has completely computerized virtually all clinical transactions, including physician orders and documentation. The VA is now integrating its clinical records across hundreds of sites of care by means of a Health Data Repository (HDR) project. ERT has been designed to provide a terminology development environment, terminology services, and maintenance services for the clinical and business content in HDR and other VHA applications. Drug, laboratory observations, and clinical document title files have been developed, and the ERT will encompass all HDR domains by 2008. Commercial tools are used to host the VHA's ERT terminology development and server environments. We will select and adopt both open-source and licensable terminology systems to provide ERT content, as well as reuse existing VA-specific terminology content. PMID- 15360842 TI - Evaluation of reliability in the use of endoscopic terminology. AB - Variability in the reporting of gastrointestinal endoscopic findings may affect the validity of analyses of data collected from clinical reports of those findings. In this project, images of 10 endoscopic findings were collected from the data repository of the Clinical Outcomes Research Initiative (CORI), all of which had been described by the reporting endoscopist. These images were presented to 52 experienced endoscopists recruited from the clinical affiliates of CORI who were asked to assign each a term from the Minimum Standard Terminology for Digestive Endoscopy. Proportion of agreement with the endoscopist varied by finding from 84.3% to 51.0% (overall 67.6% with 95% CI 63.4-71.8%). Proportion of agreement among the subjects varied by finding from 76.3% to 38.5%.(overall 55.6% with 95% CI 52.4-58.8%). Possible reasons for this lack of agreement are discussed. PMID- 15360843 TI - Challenges in precisely aligning models of human anatomy using generic schema matching. AB - This paper describes how we used generic schema matching algorithms to align the Foundational Model of Anatomy (FMA) and the GALEN Common Reference Model (CRM), two large models of human anatomy. We summarize the generic schema matching algorithms we used to identify correspondences. We present sample results that highlight the similarities and differences between the FMA and the CRM. We also identify uses of aggregation, transitivity, and reification, for which generic schema matching fails to produce an accurate mapping and present manually constructed solutions for them. PMID- 15360844 TI - Mapping Japanese medical terms to UMLS Metathesaurus. AB - This paper introduces and reports the results for a project to map Japanese medical terms to the UMLS Metathesaurus. The "Thesaurus for Medical and Health related Terms version 5" published in 2003 by the Japan Medical Abstracts Society and UMLS version 2002AC provided by NLM were used in this study. The goal was to judge the validity of the correlation between the Japanese and English terms that belong to the same MeSH concept. Fifteen medicine, nursing, and library science professionals, excluding JAMAS, used a custom designed Web interface to perform this task. About 10% of the concepts were judged as invalid, and the reasoning behind these failures were analyzed. Experience from this project can be used to estimate the manpower required to revise the Japanese thesaurus after future revisions to UMLS or MeSH. PMID- 15360845 TI - Using compound codes for automatic classification of clinical diagnoses. AB - Classification of diagnoses (a.k.a. coding) is the central part of current concept based medical IR systems. Some classification systems contain over 30,000 distinct codes which makes classifying clinical documents a time consuming labor intensive and error prone process. This paper presents a simple methodology for cleaning up and reusing existing manually coded diagnostic statements mainly extracted from clinical notes to build predictive models using a sparse-feature implementation of a Naive Bayes classifier. One of the problems addressed is that diagnostic statements often contain several diagnoses and are assigned several codes resulting in a multi-class classification problem. We investigate one possible way of addressing this problem by introducing compound (multiple code) categories. We present experimental results of classifying >16,000 randomly selected diagnostic strings into 19 top level categories. A small improvement (3%) with using compound categories over simple categories indicates that using multiple code categories is a promising solution, although clearly in need of further research and refinement. PMID- 15360846 TI - Coping with medical polysemy in the semantic web: the role of ontologies. AB - Polysemy is the linguistic phenomenon by which a term has more than one meaning. It is not a negligible issue in information management, since an effective and unambiguous sharing of the semantic content of data among different databases or knowledge repositories is needed. The paper illustrates a case of polysemy (concerning inflammation), and puts it within the framework provided by the DOLCE+ foundational ontology. This solution enables us to formally represent several senses of inflammation, and their interrelationships. This we take as a demonstration of how ontologies play an essential role in providing precisely the conceptual foundations that are needed in order to make the intended meaning of natural language expressions available to all the (artificial or human) agents that could be involved in the semantic web PMID- 15360847 TI - Problems and solutions with integrating terminologies into evolving knowledge bases. AB - We have merged two established anatomical terminologies with an evolving ontology of biological structure: the Foundational Model of Anatomy. We describe the problems we have encountered and the solutions we have developed. We believe that both the problems and solutions generalize to the integration of any legacy terminology with a disciplined ontology within the same domain. PMID- 15360848 TI - ICPC Multilingual Collaboratory: a Web- and Unicode-based system for distributed editing/translating/viewing of the multilingual International Classification of Primary Care. AB - The International Classification of Primary Care (ICPC) is a clinical classification containing 726 clinical concepts, available in over 20 languages, augmented by links to ICD-10 concepts. It is employed in clinical information systems in several European countries, Israel, Japan, and Australia. In translating ICPC, it has been challenging to manage the flow of multilingual information, maintain its quality, and optimize its portability, particularly in light of the numerous character encodings used to represent its content. The ICPC Multilingual Collaboratory (IMC) is a World Wide Web-based environment, created to allow the viewing, maintenance, and translation of ICPC content by a dispersed international editorial staff. Based upon open-source software, it represents ICPC content using the Unicode standard for character encoding. The system implements three interfaces to ICPC data: 1) a password-protected editorial interface which instantiates a hierarchical authority model and communication channels for review and control of content, including a means of up-loading new candidate translations; 2) an openly accessible read-only interface, with e-mail access to the editors (providing another level of content review); and, 3) a management interface for the system administrator. The completed system powerfully demonstrates the ability of the World Wide Web, open-source software, and Unicode to expedite and simplify international multilingual collaboration, even in a world in which Unicode support is incomplete on existing computing platforms. PMID- 15360849 TI - Mapping various information sources to a semantic network. AB - Giessen Data Dictionary Server (GDDS) provides context sensitive information services to disparate clinical applications by automatically navigating a semantic network that stores medical knowledge. Mapping multiple information sources to single clinical application becomes a challenge due to different organization of semi-structured information sources. Linking huge unstructured information sources such as medical literature is even more challenging because we need to develop a mechanism to organize unstructured information and take into account the scalability issue. In this paper, we have successfully mapped two drug information sources by developing an independent subnet for each source and linking them at proper nodes. For medical literature, we have demonstrated that the semantic network of the Unified Medical Language Systems and human assigned topics to each document can be used to organize the large amount of medical literature into the framework of the GDDS service. PMID- 15360850 TI - A corpus driven approach applying the "frame semantic" method for modeling functional status terminology. AB - In an effort to unearth semantic models that could prove fruitful to functional status terminology development we applied the "frame semantic" method, derived from the linguistic theory of thematic roles currently exemplified in the Berkeley "FrameNet" Project. Full descriptive sentences with functional-status conceptual meaning were derived from structured content within a corpus of questionnaire assessment instruments commonly used in clinical practice for functional-status assessment. Syntactic components in those sentences were delineated through manual annotation and mark-up. The annotated syntactic constituents were tagged as frame elements according to their semantic role within the context of the derived functional-status expression. Through this process generalizable "semantic frames" were elaborated with recurring "frame elements". The "frame semantic" method as an approach to rendering semantic models for functional-status terminology development and its use as a basis for machine recognition of functional status data in clinical narratives are discussed. PMID- 15360851 TI - Logical support for terminological modeling. AB - Terminological modeling, in particular in medical domains, is difficult and has enjoyed growing attention over the recent past. In practice, logical reasoning can play a fundamental role to support the modeling process. Nevertheless, the development of the logical tools is usually driven by computational or theoretical criteria rather than by the direct needs of a modeler. In this paper we attempt to shift this balance and discuss a number of logical reasoning services to support the modeler of a (medical) terminology to construct a formally sound, complete and concise terminology. This modeling support is logical as it is based on reasoning services with respect to formally defined semantics. Practical results of this systematic discussion are formal definitions of several new reasoning tasks. PMID- 15360852 TI - The role of foundational relations in the alignment of biomedical ontologies. AB - The Foundational Model of Anatomy (FMA) symbolically represents the structural organization of the human body from the macromolecular to the macroscopic levels, with the goal of providing a robust and consistent scheme for classifying anatomical entities on the basis of explicit definitions. This scheme also provides a template for modeling pathology, physiological function and genotype phenotype correlations, and it can thus serve as a reference ontology in biomedical informatics. Here we articulate the need for formally clarifying the is-a and part-of relations in the FMA and similar ontology and terminology systems. We diagnose certain characteristic errors in the treatment of these relations and show how these errors can be avoided through adoption of the formalism we describe. We then illustrate how a consistently applied formal treatment of taxonomy and partonomy can support the alignment of ontologies. PMID- 15360853 TI - A tool for abstracting relevant classes of concepts: the Common Ancestry Summarizer. AB - Controlled Medical Terminologies (CMTs) are indispensable tools for present medical information systems and medical informatics researches. Concept-oriented architecture and multiple-hierarchy have been accepted as two of the desiderata of contemporary CMTs. A common problem for informaticians working with class based methods for terminologies is to find the Most Relevant Common Ancestor (MRCA) for a group of concepts, the common ancestor which has the closest semantic distance to all the given concepts. Finding an ancestor concept to summarize a group of concepts is required to map concepts from the knowledgebase to those of appropriate granularity in CMTs. However, manually exploring the hierarchical relationships and determining the MRCA are daunting tasks due to the massive size, the multiple hierarchies and in some case the cycles of the hierarchical networks of terminologies. In our study, we developed a web-based visualization tool, the Common Ancestry Summarizer (CAS), that graphically displays the hierarchical relations of multiple concepts within CMTs, and also assist users to identify the MRCA of these concepts by identifying the Lowest Common Ancestors (LCAs). The CAS has been shown useful in organizing classes for the controlled terms used in clinical guidelines (e.g. "the bottom-up method"). PMID- 15360854 TI - Drug interaction ontology (DIO) for inferences of possible drug-drug interactions. AB - Drug Interaction Ontology (DIO) was developed for formal representation of pharmacological knowledge. It provides a fundamental framework for accumulation of reusable knowledge components in molecular pharmacology. Ontology was employed and implemented as a relational model. Some features include: 1) Drug-biomolecule interaction was assumed as a primitive knowledge element. 2) Symbolic representation was developed for drug-biomolecule interaction. Consequences of two conjugated units of interaction were defined by using symbols. These are applied for query development for identification of possible drug-drug interaction. 3) The triadic relationship model was developed as a ground model for bio-logical interactions and/or function, including semantic ones. One application of DIO is to support hypothesis generation of drug interaction by providing new hypotheses from a structured database storing literature information on known drug-biomolecule interactions. A knowledge base using DIO that contains information beginning with anti-cancer drugs is now under development. Detection of possible drug interaction was tested and its capacity to lead clinically known ones was confirmed. The system generated theoretically possible drug-drug interactions, which implies potential usefulness of new drugs to be tested before actual clinical application. In this paper, sorivudine and 5 fluorouracil mediated by dihydropyrimidine dehydrogenase are presented. PMID- 15360855 TI - Comparing associative relationships among equivalent concepts across ontologies. AB - Methods for comparing associative relationships across ontologies often rely solely on lexical similarity between the names of the relationships, which may lead to missed matches and inaccurate matches. In this paper, we propose a novel method based on the analysis of paths between equivalent concepts across ontologies. Patterns of relationships are identified for each associative relationship. The most frequent patterns indicate a correspondence between an associative relationship in one ontology and one relationship (or combination thereof) in the other. We applied this method to two ontologies of anatomy. Our method was able to identify the correspondence between relationships even in the absence of lexical similarity between relationship names. The various types of matches identified are discussed as well as the application of this method to detecting inconsistencies across the ontologies. PMID- 15360856 TI - Comparison of methods for evaluation of a medical terminological system. AB - The importance of terminological systems (TS) to support standardized and structured documentation of medical data is commonly recognized. The usability of TS in real practice strongly depends on the completeness and the correctness of the content of the TS. We here present four different methods that can be applied to evaluate a TS' content. All four methods were applied in a case study. We make a comparison of 1) the results of two methods that focus on the completeness of the content and that differ in the application of the TS that they focus on and 2) the results of an automated and a manual evaluation of the correctness of the content. Finally we summarize the results of all four methods and analyze whether they overlap or complement each other. PMID- 15360857 TI - A comparison of semantic categories of the ISO reference terminology models for nursing and the MedLEE natural language processing system. AB - Natural language processing (NLP) systems have demonstrated utility in parsing narrative texts for purposes such as surveillance and decision support. However, there has been little work related to NLP of nursing narratives. The purpose of this study was to compare the semantic categories of a NLP system (Medical Language Extraction and Encoding [MedLEE] system) with the semantic domains, categories, and attributes of the International Standards Organization(ISO) reference terminology models for nursing diagnoses and nursing actions. All but two MedLEE diagnosis and procedure-related semantic categories mapped to ISO models. In some instances, we found exact correspondence between the semantic structures of MedLEE and the ISO models. In other situations (e.g. aspects of site or location), the ISO model was not as granular as MedLEE. For clinical procedure and non-invasive examination, two ISO nursing action model components (action and target) were required to represent the MedLEE semantic category. The ISO model requires additional specification of selected semantic categories for the abstract semantic domains in order to achieve the objective of using NLP to parse and encode data from nursing narratives. Our analysis also suggests areas for extension of MedLEE. PMID- 15360858 TI - VA National Drug File Reference Terminology: a cross-institutional content coverage study. AB - BACKGROUND: Content coverage studies provide valuable information to potential users of terminologies. We detail the VA National Drug File Reference Terminology's (NDF-RT) ability to represent dictated medication list phrases from the Mayo Clinic. NDF-RT is a description logic-based resource created to support clinical operations at one of the largest healthcare providers in the US. METHODS: Medication list phrases were extracted from dictated patient notes from the Mayo Clinic. Algorithmic mappings to NDF-RT using the SmartAccess Vocabulary Server (SAVS) were presented to two non-VA physicians. The physicians used a terminology browser to determine the accuracy of the algorithmic mapping and the content coverage of NDF-RT. RESULTS: The 509 extracted documents on 300 patients contained 847 medication concepts in medication lists. NDF-RT covered 97.8% of concepts. Of the 18 phrases that NDF-RT did not represent, 10 were for OTC's and food supplements, 5 were for prescription medications, and 3 were missing synonyms. The SAVS engine properly mapped 773 of 810 phrases with an overall sensitivity (precision) was 95.4% and positive predictive value (recall) of 99.9%. CONCLUSIONS: This study demonstrates that NDF-RT has more general utility than its initial design parameters dictated PMID- 15360859 TI - Ontology-based error detection in SNOMED-CT. AB - Quality assurance in large terminologies is a difficult issue. We present two algorithms that can help terminology developers and users to identify potential areas of improvement. We demonstrate the methodology by applying the algorithms to one of the most popular terminologies, SNOMED-CT. Analysis of the results provides evidence for the thesis that both formal logical and linguistic tools should be used in the development and quality-assurance process of large terminologies. PMID- 15360860 TI - Identifying respiratory findings in emergency department reports for biosurveillance using MetaMap. AB - Clinical conditions described in patients' dictated reports are necessary for automated detection of patients with respiratory illnesses such as inhalational anthrax and pneumonia. We applied MetaMap to emergency department reports to extract a set of 71 clinical conditions relevant to detection of a lower respiratory outbreak. We indexed UMLS terms in emergency department reports with MetaMap, filtered the indexed output with a specialized lexicon of UMLS terms for the domain, and mapped the clinical conditions of interest to concepts in the lexicon. We compared MetaMap's ability to accurately identify the conditions against a physician's manual annotations and evaluated incorrectly indexed features to determine what additional processing is necessary. MetaMap identified the clinical conditions with a recall of 0.72 and a precision of 0.56. Necessary processing beyond MetaMap's indexing includes finding validation, temporal discrimination, anatomic location discrimination, finding-disease discrimination, and contextual inference. Successful identification of clinical conditions in an emergency department report with MetaMap requires processing techniques specific to the clinical question of interest. PMID- 15360861 TI - Using a terminology server and consumer search phrases to help patients find physicians with particular expertise. AB - OBJECTIVES: To design and implement a real world application using a terminology server to assist patients and physicians who use common language search terms to find specialist physicians with a particular clinical expertise. METHOD: Terminology servers have been developed to help users encoding of information using complicated structured vocabulary during data entry tasks, such as recording clinical information. We describe a methodology using Personal Health Terminology trade mark and a SNOMED CT-based hierarchical concept server. RESULTS: Construction of a pilot mediated-search engine to assist users who use vernacular speech in querying data which is more technical than vernacular. CONCLUSION: This approach, which combines theoretical and practical requirements, provides a useful example of concept-based searching for physician referrals. PMID- 15360862 TI - Overcoming barriers to evaluation of terminological systems. AB - Evaluation of terminological systems has been demonstrated to be a complicated task. This is due to the broad range of terminological systems, their application, and the clinical contexts in which they can be applied. We propose an evaluation framework that explicitly distinguishes an application-independent description of terminological systems, methods to determine application-dependent requirements, and methods to assess the applicability. In order to support a systematic application-independent description of terminological systems, we present a categorization of characteristics, including explicit questions. The answers to these questions can be mapped to the requirements for a certain application of a terminological system. This framework aims at reducing the efforts for determining which terminological system is applicable for a certain clinical setting. PMID- 15360863 TI - Translating the International Classification for Nursing Practice (ICNP)--an experience from two countries. AB - The International Classification for Nursing Practice (ICNP) has been published in the alpha, beta 1, and beta 2 versions that, together with forthcoming versions, are going to be translated into many languages. It is important that the translation process facilitates a valid and reliable translation. Experiences from two countries are given as well as recommendations for future work. There is a need for an ongoing development of the ICNP, including feedback from numerous countries to ICN. PMID- 15360864 TI - Summarization of an online medical encyclopedia. AB - We explore a knowledge-rich (abstraction) approach to summarization and apply it to multiple documents from an online medical encyclopedia. A semantic processor functions as the source interpreter and produces a list of predications. A transformation stage then generalizes and condenses this list, ultimately generating a conceptual condensate for a given disorder topic. We provide a preliminary evaluation of the quality of the condensates produced for a sample of four disorders. The overall precision of the disorder conceptual condensates was 87%, and the compression ratio from the base list of predications to the final condensate was 98%. The conceptual condensate could be used as input to a text generator to produce a natural language summary for a given disorder topic. PMID- 15360865 TI - Linguistic analysis: terms and phrases used by patients in e-mail messages to nurses. AB - While some researchers are focusing on mapping free-text within health care fields into controlled vocabularies and classifications, many researchers are focusing on consumers' vocabularies. Using natural language processing (NLP) tools, such as MetaMap, to extract and map into terms in a controlled vocabulary is one way of understanding the pattern of terms used by lay people. Before an NLP tool can be effectively and efficiently used to extract concepts and create machine-understandable interpretations of the data, the appropriateness of the tool needs to be determined. This study aims were to determine the appropriateness of linguistic meaning captured for terms and phrases used by patients in electronic mail messages to nurses, using nursing-specific MetaMap output. Twenty messages were randomly selected from the 241 messages data set. Results indicated that four out of six nursing classification systems captured more than 50 % of the parsed word's linguistic meaning. This study demonstrates that it is possible to automatically extract and capture the linguistic meaning of the terms patients use in their electronic mail messages. PMID- 15360866 TI - A surgeon can operate or approach a complicated integral calculus instead of a renal one: implications for conceptual and lexical semantic disambiguation. AB - This paper approaches lexical semantic disambiguation and polysemy in the context of medical language understanding. These issues have been addressed as linguistic and cognitive requirements to build a knowledge extraction tool that turns natural language input into conceptual graphs. Previous results obtained with semantic analysis of medical terms in the domain of transplantation and organ failure prompt us to check the capabilities of our prototype to deal with ambiguities and polysemy. Starting from linguistic observation, we attempt to demonstrate how the respect of ambiguities when the co-text is not sufficient for disambiguation implies to introduce a void type in the concept type lattice. Then we show how the "creative use of words" (i.e. new senses in novel context) imposes to dynamically allocate type, categories and roles with the co-text. PMID- 15360867 TI - Modeling, the essential step to consolidate and integrate a national NMDS. AB - How to develop a nursing minimum data set (NMDS) as a part of a nursing information system, which is itself a part of a health information system, this is the challenge presented in this paper and realized in Switzerland through the NURSING data project. The development of the CH-NMDS is not just a selection of data but a real conceptual work consisting in making the link between the information system and the terminology system and this for any level of aggregation. The building of a conceptual coherence between the collection of the data, their storage and their utilization for decision making was made by using the object modeling language, not for programming aims but statistical purposes. For example, the nursing phenomena and the nursing interventions are considered as classification concepts as well as sets of data. PMID- 15360868 TI - Representing public health nursing intervention concepts with HHCC and NIC. AB - PURPOSE: It is imperative that public health nurses define their services and provide evidence supporting the effectiveness of interventions. The purpose of this paper is to examine the ex-tent to which two standardized nursing terminologies--Home Health Care Classification (HHCC) and Nursing Interventions Classification (NIC)--represent public health nursing practice according to core public health function in Public Health Nursing Intervention model. METHODS: First, we divided all HHCC and NIC interventions into intervention focus levels: individual/family-focused, community-focused, and system-focused. Second, we categorized HHCC and NIC interventions according to core public health functions: assessment, policy development, and assurance and the categories of interventions in the PHI Model. RESULTS: We identified HHCC and NIC Nursing interventions that represented public health nursing concepts across core public health functions and categories of the PHI model. Analysis of the findings demonstrated that HHCC and NIC have terms for the concepts in the PHI model. CONCLUSION: Although HHCC and NIC cover many concepts in public health nursing practice, additional research is needed to extend these terminologies and to evaluate other standardized terminologies that can reflect more comprehensively public health nursing interventions. PMID- 15360869 TI - MeSH and specialized terminologies: coverage in the field of molecular biology. AB - There is real need among researchers for data in molecular biology, due to the explosive development in this field. Information extraction techniques have been developed in this specific domain, to extract structured information from the literature. The National Library of Medicine provides access to 12 million MEDLINE citations, accessible via PubMed. Information retrieval techniques which are used to select documents that are relevant according to user request. As other domains in medicine, biomolecular scientific literature is indexed by MeSH terms. Automatic identification of protein and gene names in scientific articles is an important step in the indexing process, since proteins and genes are often associated with multiple names, and authors alternate between these synonyms. In this article we assess the contribution of specialized terminologies in the field of molecular biology compared to MeSH. PMID- 15360870 TI - Acquiring meaning for French medical terminology: contribution of morphosemantics. AB - Morphologically complex words, and particularly neoclassical compounds, form more than 60% of the neologisms in the biomedical field. Guessing their definitions and grouping them into semantic classes by means of lexical relations are thus two crucial improvements for handling these words, e.g., for information retrieval, indexing and text understanding applications. This paper describes a morphosemantic linguistic-based parser called DeriF, currently developed in the framework of two projects, UMLF and VUMeF, and its application to French biomedical derived and compound words. It shows how the resulting morphologically tagged lexicon is enriched by semantic relations leading both to the synthesis of pseudo-definitions and to the constitution of classes of synonyms, hypo- and hypernyms. PMID- 15360871 TI - Evaluation of SNOMED coverage of Veterans Health Administration terms. AB - Veterans Health Administration (VHA) is now evaluating use of SNOMED-CT. This paper reports the first phase of this evaluation, which examines the coverage of SNOMED-CT for problem list entries. Clinician expressions in VA problem lists are quite diverse compared to the content of the current VA terminology Lexicon. We selected a random set of 5054 narratives that were previously "unresolved" against the Lexicon. These narratives were mapped to SNOMED-CT using two automated tools. Experts reviewed a subset of the tools' matched, partly matched, and un-matched narratives. The automated tools produced exact or partial matches for over 90% of the 5054 unresolved narratives. SNOMED-CT has promise as a coding system for clinical problems. In subsequent studies, VA will examine the coverage of SNOMED for other clinical domains, such as drugs, allergies, and physician orders. PMID- 15360872 TI - Terminology access methods leveraging LDAP resources. AB - Health terminologies have become more complex, more massive, and more ubiquitous in the modern healthcare enterprise. Present technology makes the use of these terminologies by humans, unaided by machines, virtually impossible. However, system and message interoperability can be severely compromised if the software services deploying terminology content and interfaces are themselves non standard. We review some characteristics for good terminology services and introduce an open-source, robust, widely deployed and widely available software resource to underpin terminology service implementations. The Lightweight Directory Access Protocol, or LDAP, is compared with alternative technologies. We describe a reference implementation of terminology services built around the HL7 Common Terminology Services using LDAP methods. We propose that LDAP is well suited as a common platform for federated, synchronized, and algorithmically distributed terminology content from multiple sources. PMID- 15360873 TI - Medical data abstractionism: fitting an EMR to radically evolving medical information systems. AB - Growing and maintaining a simple and flexible EMR (Electronic Medical Record) becomes a complicated task in light of diverse and distributed legacy data representation, advancing technologies, changes in medical practice and procedure, and changes in data regulation. Utilizing several abstraction mechanisms can simplify application development and maintenance, and provide flexibility for data evolution and migration. Newer applications built on these abstractions can be the beneficiary of slower obsolescence and lower maintenance costs. PMID- 15360874 TI - Automating terminological networks to link heterogeneous biomedical databases. AB - As cross-disciplinary research escalates, researchers are facing the challenge of linking disparate biomedical databases that have been developed without common indexes. Manually indexing these large-scale databases is laborious and often impractical. Solutions involving mediating terminologies have been proposed, but coordination of terms from the databases of interest to these mediating terminologies is also laborious, and regular synchronization between indexes is an additional problem. In this study we describe a novel method of linking heterogeneous databases using terminology networks constructed with automated mapping methods. Linkage was established between two disparate biomedical databases (SNOMED-CT and HDG), using two relevant intermediating databases (UMLS and OMIM). One gold standard of 514 distinct matches is used as proof-of principle. In conclusion, as hypothesized, 1) Manually curated pathways provide high precision, but offer low recall, 2) the automated terminology pathways can significantly increase recall at acceptable precision. Taken together, our conclusion may suggest the combined manual and automated terminology networks could offer recall and precision in an incremental manner PMID- 15360875 TI - Really, is medical sublanguage that different? Experimental counter-evidence from tagging medical and newspaper corpora. AB - We compare the performance of two part-of-speech taggers trained on a German newspaper corpus for mixed types of medical documents. TnT, a tagger based on a statistical language model, outperforms Brill's rule-based tagger, and supplied with additional lexicon resources matches state-of-the-art performance figures (close to 97% accuracy) on the medical corpus. We explain this unexpected result by focusing on the statistically significant part-of-speech type overlap between the newspaper training set and the medical test set. At least at that level, sublanguage differences seem to vanish. Thus, statistical off-the-shelf part-of speech taggers can immediately be reused for medical language processing PMID- 15360876 TI - Facilitating cancer research using natural language processing of pathology reports. AB - Many ongoing clinical research projects, such as projects involving studies associated with cancer, involve manual capture of information in surgical pathology reports so that the information can be used to determine the eligibility of recruited patients for the study and to provide other information, such as cancer prognosis. Natural language processing (NLP) systems offer an alternative to automated coding, but pathology reports have certain features that are difficult for NLP systems. This paper describes how a preprocessor was integrated with an existing NLP system (MedLEE) in order to reduce modification to the NLP system and to improve performance. The work was done in conjunction with an ongoing clinical research project that assesses disparities and risks of developing breast cancer for minority women. An evaluation of the system was performed using manually coded data from the research project's database as a gold standard. The evaluation outcome showed that the extended NLP system had a sensitivity of 90.6% and a precision of 91.6%. Results indicated that this system performed satisfactorily for capturing information for the cancer research project. PMID- 15360877 TI - The oncological nurse assistant: a web-based intelligent oncological nurse advisor. AB - As more people get cancer the need for medical guidance increases. In Norway, one of the providers of medical guidelines is the Norwegian Cancer Association where oncological nurses assist people with a cancer diagnosis or their relatives. The nurses search through both national and internal guide-books and web pages. The input to this process is mostly dis-charge letters. The whole process is time consuming. To serve more patients, PaSent, a web-based intelligent oncological nurse advisor, has been developed. Through using discharge letters as input to our neural network based information retrieval system PaSent, we have been able to provide relevant medical information to the patient as well as to the health personnel themselves. The PaSent search method uses predefined knowledge about the context, paired with the vocabulary of the input document, to compute a relevance measure for a potential result document. The system has been validated by oncological nurses and medical doctors. In the reported experiments, the achieved search results from PaSent have been comparable to the results achieved by the health personnel. PMID- 15360878 TI - Characteristics of outpatient clinical decision support systems: a taxonomic description. AB - Computer-based clinical decision support systems (CDSSs) have been championed for their potential to improve health-care quality. However, there has been no systematic study of the types of CDSSs that have been developed. In previous work, we developed the CDSS Taxonomy for comprehensively describing the technical, workflow, and contextual characteristics of CDSSs. We now use the CDSS Taxonomy to describe outpatient CDSSs evaluated in randomized controlled trials published between 1998 and 2002. 31 studies comprising 42 CDSS systems were included in our analysis. The majority of systems used rule-based reasoning engines to "push" explicit, individualized recommendations concerning non-urgent decisions to clinicians or patients, but not both. 71% of the systems required someone to manually enter data into the system or to process the system output for use by the target decision maker. The average kappa for coding agreement was > 0.6. Our findings demonstrate that outpatient CDSSs vary greatly in design and function. Many impose a data entry or output-processing burden on clinic staff. More complete reporting of CDSS characteristics is needed in the literature. PMID- 15360879 TI - Impact of patient feedback on residents' handheld computer use: a multi-site study. AB - Handheld computers are being proposed as a resource for ambulatory care, yet physician reluctance to use the devices in the presence of patients is reported to be a barrier to optimal use. This study examined patient attitudes toward the use of handheld computers and the impact of those attitudes on the physicians' use of the handheld computers in the patient care setting. Most patients hold favorable attitudes toward physician use of handheld computers. Providing these data to physicians, however, did not uniformly lead to significant increases in handheld use in the presence of the patient. More understanding of the factors that influence behavior change and methods that will decrease barriers to the use of handhelds is needed PMID- 15360880 TI - Automated medical problem list generation: towards a patient timeline. AB - The problem-oriented electronic medical record has been investigated as an alternative to source-oriented organization of patient data. At the core of a problem-oriented view is the medical problem list. Maintenance of the medical problem list is often manual, making it highly user dependent. We detail the beginnings of an automated medical problem list generator based on ICD-9: given a set of ICD-9 codes associated with a patient record, the system maps the codes (and related data) to an anatomy-centric hierarchy. 1 million patient encounters from an outpatient setting were reviewed to generate a unique set of 7,890 ICD-9 codes. Natural language processing of the ICD-9 string descriptions identified 1,981 anatomical terms, which were subsequently mapped to one of 21 anatomical categories. The output of the medical problem list generator was then used to create a problem-oriented, gestalt view of a patient's medical record. Preliminary evaluation of the generator revealed 100% recall, but only 60% precision. This initial work has highlighted several issues in defining a medical problem list, including questions of granularity and performance trade-offs. PMID- 15360881 TI - The economics of integrated electronic medical record systems. AB - The decision to adopt electronic medical record systems in private practices is usually based on factors specific to the practice--the cost, cost and timesaving, and impact on quality of care. As evident by the low adoption rates, providers have not found these evaluations compelling. However, it is recognized that the widespread adoption of EMR systems would greatly benefit the health care system as a whole. One explanation for the lack of adoption is that there is a misalignment of the costs and benefits of EMR systems across the health care system. In this paper we present an economic model of the adoption of EMR systems that explicitly represents the distribution of costs and benefits across stakeholders (physicians, hospitals, insurers, etc.). We discuss incentive systems for balancing the costs and benefits and, thus, promoting the faster adoption of EMR systems. Finally, we describe our plan to extend the model and to use real-world data to evaluate our model. PMID- 15360882 TI - Context in care--requirements for mobile context-aware patient charts. AB - The hospital ward is a highly dynamic work environment, in which healthcare personnel rapidly switch from one task to another. The process is partly planned, and partly driven by events and interrupts. A mobile electronic patient chart (MEPC) will be an important tool for supporting order entry and accessing, communicating, and recording clinical information. The users need to switch from one context to another with minimal delay and effort. Context-awareness, the ability to sense relevant situational information, can allow the user interface of the MEPC to adapt to various situations. In this paper, we present a future scenario from the coronary care unit. This scenario is analyzed and discussed in order to develop requirements for design methods, context models, and system properties of the MEPC. PMID- 15360883 TI - Organizing literature information for clinical decision support. AB - Answers to clinical questions occurring during healthcare practitioner/patient interaction can be often found in National Library of Medicine's (NLM) databases. The recent advances in wireless handheld computers promise to make them a widely used tool to deliver needed information to the practitioner at the point of service. This paper addresses challenges in organizing and presenting information obtained from NLM's MEDLINE database of indexed citations in a way that will help practitioners reduce literature search time on handheld computers. We study two clustering algorithms and two methods of labeling document clusters. PMID- 15360884 TI - Automated clinical annotation of tissue bank specimens. AB - Modern, molecular bio-medicine is driving a growing demand for extensively annotated tissue bank specimens. With careful clinical, pathologic and outcomes annotation, samples can be better matched to the research question at hand and experimental results better understood and verified. However, the difficulty and expense of detailed specimen annotation is well beyond the capability of most banks and has made access to well documented tissue a major limitation in medical re-search. In this context, we have implemented automated annotation of banked tissue by integrating data from three clinical systems--the cancer registry, the pathology LIS and the tissue bank inventory system--through a classical data warehouse environment. The project required modification of clinical systems, development of methods to identify patients between and map data elements across systems and the creation of de-identified data in data marts for use by researchers. The result has been much more extensive and accurate initial tissue annotation with less effort in the tissue bank, as well as dynamic ongoing annotation as the cancer registry follows patients over time. PMID- 15360885 TI - Design of a standards-based external rules engine for decision support in a variety of application contexts: report of a feasibility study at Partners HealthCare System. AB - This project explored functional requirements for an institution-wide method, at Partners HealthCare, for interpreting clinical knowledge for decision support. Such knowledge is currently incorporated in a variety of clinical applications, yet the methods of representation and of execution vary and the ability to author/edit the rules by human experts is limited. We expanded on a 2002 "Knowledge Inventory" at Partners to evaluate feasibility of designing a single representation approach entailing: (a) exploration of specific needs of different applications, in terms of kinds of response required (synchronous/asynchronous, time criticality, etc.), context (e.g., implied patient, time frame, or episode), and kinds of actions to be triggered; (b) kind of representation of knowledge and feasibility of casting knowledge in the form of if em leader then statements; and (c) data and knowledge resources used (implied data model, and particular knowledge sources and terminology sources). The result of analysis was to design an architecture to accomplish this goal. We also did preliminary analysis of requirements for authoring for such a representation, and for implementation. PMID- 15360886 TI - A comparative study on concept representation between the UMLS and the clinical terms in Korean medical records. AB - The Unified Medical Language System (UMLS) is a rich source of knowledge in the biomedical domain. In this paper, we evaluated the coverage of UMLS as compared with Korean medical terms and identified differences in concept representation between two vocabulary sets. We measured the concept coverage by mapping clinical terms extracted from the discharge records of Seoul National University Hospital (SNUH) and the UMLS "Sign or Symptom" and "Disease or Syndrome" concepts. Thirty five percent of the complaints in the SNUH were conceptually matched with the UMLS "Sign or Symptom" concept. Fifty-eight percent were found to be matched with the UMLS "Disease or Syndrome" concept rather than the "Sign or Symptom" concept. The remaining seven percent were not found in the UMLS concepts mapped above or those terms that used to special circumstances of a tertiary hospital in Korea. We then analyzed some of different expression patterns used by the two vocabulary sets and addressed issues to be taken into consideration. PMID- 15360887 TI - Requirements of tools and techniques to support the entry of structured nursing data. AB - The benefits of structured data are widely accepted within the nursing informatics community. However, despite the existence of structured data in the form of well-established nursing terminologies, computer-based nursing record systems are yet to achieve widespread adoption and few of the potential benefits have yet to be realized. In this paper we argue the need for tools and techniques to support the entry of structured nursing data into computer-based systems. In the absence of a generally accepted solution, we build on preliminary work carried out at the 2002 Nursing Terminology Summit and analyze the results of other studies in order to identify a preliminary set of requirements or desiderata for such tools and techniques. These requirements are centered on: how structured data is presented to users for selection; how to mediate between a variety of conceptual structures--terminologies, information models, user interface models and models of the clinical process; and how to reduce the considerable modeling burden through reuse of modeling constructs. Further applied research is needed with the ultimate goal of developing a general solution that will benefit nurses, other professionals and ultimately their patients. PMID- 15360888 TI - A knowledge based approach for automated signal generation in pharmacovigilance. AB - BACKGROUND: Pharmacovigilance experts detect new adverse drug reactions (ADR) by manually reviewing spontaneous reporting systems. Automated signal generation aims to focus the attention of experts on drug-adverse event associations which are disproportionally present in the database. Although adverse events are coded by means of controlled vocabularies such as the MedDRA dictionary, this semantic information is not taken into account for signal generation. OBJECTIVE: To improve the performance of current signal detection algorithms using knowledge based approach. METHOD: We developed a formal ontology of ADRs and built a data mining tool that uses description logic representations of MedDRA terms to group medically related case reports. RESULTS: This knowledge based approach increased the sensitivity of signal detection with no decrease in specificity. DISCUSSION: A knowledge based approach improved the performance of signal detection tools. However, the huge work-load involved in the knowledge engineering step limits the use of this approach for machine learning. PMID- 15360890 TI - Informed consent to the secondary use of EHRs: informatic rights and their limitations. AB - It is frequently asserted that the secondary use of data contained in electronic health records (EHRs) requires the informed consent of the subjects of these records. This paper argues that while correct in principle, this has four important ethically based limitations: when it seriously threatens the equal and competing right of others, when it undermines the ability of health care providers to carry out their mandates, when it imperils the possibility of establishing and maintaining a health care system in the first place, and when it makes bona fide research impossible. Nevertheless, as a general rule, when consent can be had, it should be had. This paper provides a theoretical basis for these claims by looking at the nature and role of EHRs and provides some possible approaches for incorporating them into the health care delivery process. PMID- 15360889 TI - Dealing with organizational change when implementing EHR systems. AB - Over the past years researchers at Aalborg University have been developing a method for examining change readiness of hospital staff during the implementation of new IT-systems. (CRR; Change readiness-research). The aim is to provide the hospital-management with a tool that will lead to an optimal implementation of new IT-systems. The method has recently been used in department Y at Skejby Hospital, Skejby, Denmark. 81% out of 241 employees answered the distributed questionnaire, and the results showed, that the employees have some expectations to the new EHR-system, which are important to either confirm or reject; on the other hand a great part of the employees lack basic computer skills. Thus the CRR method has provided the Organization with important information before the implementation of the new EHR-system. PMID- 15360891 TI - CliniViewer: a tool for viewing electronic medical records based on natural language processing and XML. AB - With the evolving use of computers in healthcare, the electronic medical record (EMR) is becoming more and more popular. A tool is needed that would enable physicians to accurately and efficiently access clinical information in multiple medical records associated with a particular patient. Both natural language processing (NLP) and the eXtensible Markup Language (XML) have been used in the clinical domain for capturing, representing, and utilizing clinical information and both have shown great potential. In this paper, we demonstrate another use of XML and NLP through CliniViewer, a tool that organizes and presents the clinical information in multiple records. We also describe the flexibility and capability provided when combining XML and NLP to summarize, navigate, and conceptualize structured information. The tool has been fully implemented and tested using patients with multiple discharge summaries. PMID- 15360892 TI - Clinical quality needs complex adaptive systems and machine learning. AB - The vast increase in clinical data has the potential to bring about large improvements in clinical quality and other aspects of healthcare delivery. However, such benefits do not come without cost. The analysis of such large datasets, particularly where the data may have to be merged from several sources and may be noisy and incomplete, is a challenging task. Furthermore, the introduction of clinical changes is a cyclical task, meaning that the processes under examination operate in an environment that is not static. We suggest that traditional methods of analysis are unsuitable for the task, and identify complexity theory and machine learning as areas that have the potential to facilitate the examination of clinical quality. By its nature the field of complex adaptive systems deals with environments that change because of the interactions that have occurred in the past. We draw parallels between health informatics and bioinformatics, which has already started to successfully use machine learning methods. PMID- 15360893 TI - Evaluation of KNAVE-II: a tool for intelligent query and exploration of patient data. AB - We present the results of a preliminary evaluation of KNAVE-II, a distributed knowledge-based computational framework for visualization, interpretation, and exploration of longitudinal clinical data and of multiple levels of concepts derivable from these data. KNAVE-II uses a distributed architecture to access at run-time clinical time-oriented data, a domain-specific knowledge base containing properties of the clinical data, and a knowledge-based problem-solving method for computing on-the-fly interpretations of these data. The purpose of the evaluation was to compare efficiency and user satisfaction when answering clinical queries of variable complexity about clinical time-oriented data using KNAVE-II, versus using methods available in standard clinical settings: paper chart or electronic spreadsheet (ESS). Subjects answered high-complexity queries significantly faster using KNAVE-II than when using paper or ESS. User satisfaction with KNAVE-II was significantly superior compared to satisfaction using paper or ESS, based on a standard usability scale. Users also explicitly ranked KNAVE-II as superior to paper and the ESS. PMID- 15360894 TI - Automatic generation of repeated patient information for tailoring clinical notes. AB - Dictating clear, readable, and accurate clinical notes can be a time-consuming task for physicians. Clinical notes often contain information concerning the patient's medical history and current medical condition which is propagated from one clinical note to all follow-up clinical notes for the same patient. In this paper, we present a system which, given a clinical note, automatically determines what information should be repeated, and then generates this information for the physician for a new clinical note. We use semantic patterns for capturing the rhetorical category of sentences, which we show to be useful for determining whether the sentence should be repeated. Our system is shown to perform better than a baseline metric based on precision/recall results. Such a system would allow clinical notes to be more complete, timely, and accurate. PMID- 15360895 TI - Electronic medical records in solo/small groups: a qualitative study of physician user types. AB - BACKGROUND: There is growing recognition that physician use of electronic medical records (EMRs) is critical for improving quality of care in outpatient settings. METHODS: We inter-viewed EMR physician champions from 20 solo/small group practices to understand different types of EMR users and their EMR-related costs and benefits. RESULTS: Interviewees differed greatly in the EMR-related benefits they generated. These differences were associated with how they used the EMR, and the amount of effort they invested in making changes to complement EMR use. We defined five types of physician EMR users: Viewers, Basic Users, Strivers, Arrivers, and System Changers. The majority of interviewees were Strivers and Arrivers, physicians who have already invested substantial time in numerous process changes that help generate EMR-related benefits. CONCLUSIONS: Incentives and comprehensive support services for facilitating complementary process changes could be important for moving physicians from one user group to another. Additional research is needed to verify this user typology and to further define the relationship between user types and EMR-generated financial and quality benefits. PMID- 15360896 TI - Implementation and evaluation of a negation tagger in a pipeline-based system for information extract from pathology reports. AB - We have developed a pipeline-based system for automated annotation of Surgical Pathology Reports with UMLS terms that builds on GATE--an open-source architecture for language engineering. The system includes a module for detecting and annotating negated concepts, which implements the NegEx algorithm--an algorithm originally described for use in discharge summaries and radiology reports. We describe the implementation of the system, and early evaluation of the Negation Tagger. Our results are encouraging. In the key Final Diagnosis section, with almost no modification of the algorithm or phrase lists, the system performs with precision of 0.84 and recall of 0.80 against a gold-standard corpus of negation annotations, created by modified Delphi technique by a panel of pathologists. Further work will focus on refining the Negation Tagger and UMLS Tagger and adding additional processing resources for annotating free-text pathology reports. PMID- 15360897 TI - Transforming clinical dictation into structured notes in an ambulatory care practice. AB - Structured clinical encounter notes offer many advantages compared to free text notes. However, cost and other issues limit their widespread use. At our institution, the electronic medical record (EMR) used by an Internal Medicine practice was designed years ago to utilize structured data. This paper describes a technique to improve the process by which structured notes are dictated and transcribed. We outsourced transcription services and then passed the files through parsing software to structure the data to the granularity required for our database. Software was built to handle all aspects of the process including file validation, parsing, and review. During our pilot project, more than 10,000 notes were parsed, reviewed and uploaded into the EMR. PMID- 15360898 TI - A careflow management system for chronic patients. AB - The management of chronic patients is a complex process, which requires the cooperation of all primary care professionals and their interaction with specialists, laboratories and personnel of different organizations. In this paper we show how a Careflow Management System (CfMS) may represent an essential component of an innovative Health Information System (HIS) able to handle the information and communication needs underlying chronic diseases management. On the basis of a general architecture designed for chronic diseases, we describe a CfMS implementation in the area of diabetes management; such a system embeds EPR and telemedicine functionalities as end-users applications as well as a module for inter-organizational communication based on contracts and on XML messages. PMID- 15360899 TI - "Knowledge management" for health: what "tools" can improve the performance of workgroups, clinicians and patients? AB - This paper is a vision paper presenting a framework that describes a set of integrated "tools" for supporting "knowledge management". These "tools" span technical, organizational and human story telling. The framework is based on the authors' involvement in many research projects, the literature and conversations sounding the issues. PMID- 15360900 TI - Generating complex clinical documents using structured entry and reporting. AB - Structured entry and reporting in medicine remains an elusive goal. Poor adoption of clinical structured entry for documentation results in part from the inherent complexity of entering patient histories, which are generally unstructured. The authors have developed a structured entry tool that has been adopted by practicing physicians for documentation of clinical encounters. To evaluate the impact of this tool on clinical documentation, the authors have performed two comparative studies investigating note complexity. Authors compared documents generated with a standard dictation/transcription model with documents generated with structured entry. Overall, documents generated with the structured entry and reporting tool contained 64% more concepts (P<0.01) than dictated documents while maintaining the same complexity. Depth and complexity of documentation with the structured entry and reporting tool varied by clinician user and by note sub section. PMID- 15360901 TI - Generic computer-based questionnaires: an extension to OpenSDE. AB - Development of computer-based questionnaires (CQs) has been an ongoing challenge since the 1960s. The added value of such CQs for data collection and the acceptance by patients have been well documented. Many questionnaire projects, however, were temporary due to dedicated software, limited funding, and lack of integration with medical information systems. Also, the use of a fixed dedicated database makes integration cumbersome, as change in content requires change to the data model. Since much of the functional requirement of CQs is not dependent on content, the challenge is to both separate functionality and database structure from content. Following these principles, we extended OpenSDE, a generic application for structured data entry, with a tool to construct and run CQs as an alternative way of data input. We propose the combination of a generic building tool and a content-independent data model as an effective strategy to tackle the above-mentioned problems in CQ development. PMID- 15360902 TI - Specifications and implementation of a new exchange format to support computerized consensus in pathology. AB - In the pathology domain, consensus sessions around multi-headed microscopes enhance reproducibility and can reduce inter- and intra-observer variability. Computerized tools and Web technology could facilitate the organization of consensus sessions and assist pathologists to agree on features that are relevant to diagnosis. In the context of the IDEM project, whose aim is to achieve a computerized platform to allow pathologists to derive consensual diagnostic during Internet-based collaborative sessions, we propose a new extension of the existing TELESLIDE format. This new extended format enables the storage and the exchange of multi-experts descriptions that will be processed by the IDEM consensus engine to produce consensual descriptions. We describe this new format and its implementation in the IDEM teleconsensus platform. PMID- 15360903 TI - Synchronized time-motion study in the emergency department using a handheld computer application. AB - Emergency departments present a challenging environment for workflow evaluation and measurement of the effect of information system interventions. This paper describes a time-motion data collection tool built as a handheld computer application. The tool allows mobility of the observer and the intuitive inter face supports rapid selection of tasks and frequent changes in tasks. It also allows documentation of overlapping tasks, a common occurrence in the emergency department. Synchronized data collection across the different providers of care in the emergency department is demonstrated, resulting in a data set from which task/event sequences can be built, documenting the ED care of individual patients. This technique should provide a very useful tool in measuring the effect of process changes in this difficult-to-assess environment, particularly changes in the information system. PMID- 15360904 TI - Predicting the impact of a new information system on workflow using a discrete event simulation. AB - Computer simulation of an information system prior to its implementation can predict time and workflow changes in a hospital department, while offering a common ground of communication across various levels in the organization. Often, the simulation can predict unexpected effects of changes to the work environment and allow experimentation with alternative scenarios at minimal cost to the department or the organization. In this paper we describe a discrete-event simulation experiment that predicted an unexpected increase in routine specimen processing time with the introduction of an information system in the HLA tissue typing lab at a major transplant center. The computer simulation enabled the reallocation of existing staff prior to the system implementation. PMID- 15360905 TI - A novel algorithm for scalable and accurate Bayesian network learning. AB - Bayesian Networks (BN) is a knowledge representation formalism that has been proven to be valuable in biomedicine for constructing decision support systems and for generating causal hypotheses from data. Given the emergence of datasets in medicine and biology with thousands of variables and that current algorithms do not scale more than a few hundred variables in practical domains, new efficient and accurate algorithms are needed to learn high quality BNs from data. We present a new algorithm called Max-Min Hill-Climbing (MMHC) that builds upon and improves the Sparse Candidate (SC) algorithm; a state-of-the-art algorithm that scales up to datasets involving hundreds of variables provided the generating networks are sparse. Compared to the SC, on a number of datasets from medicine and biology, (a) MMHC discovers BNs that are structurally closer to the data-generating BN, (b) the discovered networks are more probable given the data, (c) MMHC is computationally more efficient and scalable than SC, and (d) the generating networks are not required to be uniformly sparse nor is the user of MMHC required to guess correctly the network connectivity PMID- 15360906 TI - An application of one-class support vector machine to nosocomial infection detection. AB - Nosocomial infections (NIs)---those acquired in health care settings---are among the major causes of increased mortality among hospitalized patients. They are a significant burden for patients and health authorities alike; it is thus important to monitor and detect them through an effective surveillance system. This paper describes a retrospective analysis of a prevalence survey of NIs done in the Geneva University Hospital. Our goal is to identify patients with one or more NIs on the basis of clinical and other data collected during the survey. In this two-class classification task, the main difficulty lies in the significant imbalance between positive or infected (11%) and negative (89%) cases. To cope with class imbalance, we investigate one-class SVMs which can be trained to distinguish two classes on the basis of examples from a single class (in this case, only "normal" or non infected patients). The infected ones are then identified as "abnormal" cases or outliers that deviate significantly from the normal profile. Experimental results are encouraging: whereas standard 2-class SVMs scored a baseline sensitivity of 50.6% on this problem, the one-class approach increased sensitivity to as much as 92.6%. These results are comparable to those obtained by the authors in a previous study on asymmetrical soft margin SVMs; they suggest that one-class SVMs can provide an effective and efficient way of overcoming data imbalance in classification problems. PMID- 15360907 TI - Development and representation of a fall-injury risk assessment instrument in a clinical information system. AB - The potential for informatics solutions to address inpatient safety issues is significant; however, several challenges are associated with the development of patient safety related informatics applications. These challenges include: 1) the identification and/or development of valid and reliable instruments; 2) adequate representation of key safety concepts, constructs, and associated concepts in the clinical information system; and 3) identification of data sources for instrument pre-population. As part of a larger project aimed at identifying and addressing the information needs of clinicians while using a clinical information system, an electronic fall and injury risk assessment instrument is in development to address a hospital-based fall and injury prevention initiative. The concepts contained in the instrument are well represented by Clinical LOINC and the UMLS. Associated concepts have been identified in the existing clinical information system data dictionary for pre-population of the instrument. PMID- 15360908 TI - Using computational modeling to improve patient care unit safety and quality outcomes. AB - As part of ongoing research to investigate the impact of patient characteristics, organization characteristics and patient unit characteristics on safety and quality outcomes, we are using a computational modeling program, OrgAhead, to model patient care units' achievement of patient safety (medication errors and falls) and quality outcomes. We tuned OrgAhead using data we collected from 16 units in 5 hospitals. Subsequent validation studies demonstrated acceptable levels of correspondence between actual and virtual patient units. In this paper, we report on our initial efforts to use OrgAhead to develop testable hypotheses about the kinds of innovations that nurse managers might realistically implement on their patient care units to improve quality and safety outcomes. Our focus is on unit-level innovations that are likely to be easier for managers to implement. For all but the highest performing unit (for which we encountered a ceiling effect), we were able to generate practical strategies that improved performance of the virtual units by 6-8 percentage points. Nurse Managers have responded enthusiastically to the additional decision support for quality improvement PMID- 15360909 TI - Causal discovery using a Bayesian local causal discovery algorithm. AB - This study focused on the development and application of an efficient algorithm to induce causal relationships from observational data. The algorithm, called BLCD, is based on a causal Bayesian network framework. BLCD initially uses heuristic greedy search to derive the Markov Blanket (MB) of a node that serves as the "locality" for the identification of pair-wise causal relationships. BLCD takes as input a dataset and outputs potential causes of the form variable X causally influences variable Y. Identification of the causal factors of diseases and outcomes, can help formulate better management, prevention and control strategies for the improvement of health care. In this study we focused on investigating factors that may contribute causally to infant mortality in the United States. We used the U.S. Linked Birth/Infant Death dataset for 1991 with more than four million records and about 200 variables for each record. Our sample consisted of 41,155 re-cords randomly selected from the whole dataset. Each record had maternal, paternal and child factors and the outcome at the end of the first year--whether the infant survived or not. Using the infant birth and death dataset as input, BLCD out-put six purported causal relationships. Three out of the six relationships seem plausible. Even though we have not yet discovered a clinically novel causal link, we plan to look for novel causal pathways using the full sample. PMID- 15360910 TI - Comparison of machine learning techniques with classical statistical models in predicting health outcomes. AB - Several machine learning techniques (multilayer and single layer perceptron, logistic regression, least square linear separation and support vector machines) are applied to calculate the risk of death from two biomedical data sets, one from patient care records, and another from a population survey. Each dataset contained multiple sources of information: history of related symptoms and other illnesses, physical examination findings, laboratory tests, medications (patient records dataset), health attitudes, and disabilities in activities of daily living (survey dataset). Each technique showed very good mortality prediction in the acute patients data sample (AUC up to 0.89) and fair prediction accuracy for six year mortality (AUC from 0.70 to 0.76) in individuals from epidemiological database surveys. The results suggest that the nature of data is of primary importance rather than the learning technique. However, the consistently superior performance of the artificial neural network (multi-layer perceptron) indicates that nonlinear relationships (which cannot be discerned by linear separation techniques) can provide additional improvement in correctly predicting health outcomes. PMID- 15360911 TI - Comparison of proportional hazard model and neural network models in a real data set of intensive care unit patients. AB - There has been increased interest in using neural network model (NNM) for prognosis tasks. However, the performance of NNM has seldom been compared with that of traditional statistical models such as proportional hazard model (PHM) in real data sets. We conducted a comparative study of PHM and two types of NNM, that is, aggregate single point model (ASPM) and multiple point model (MPM), using a real data set of intensive care unit patients. The three models were developed using the 70% training subset and their predictive accuracy were assessed using the 30% testing subset according to classification accuracy, area under receiver operating curve, and concordance index. Overall, the highest predictive accuracy was found in MPM, followed by PHM and ASPM. MPM is likely to have the potential ability to provide more accurate estimation of prognosis than PHM and ASPM PMID- 15360912 TI - Virtual interactive practice: utilizing healthcare information systems to contextualize the skills associated with clinical decision making within nurse education. AB - This paper reports on a Virtual Interactive Practice (VIP) project that has the potential to revolutionise the educational delivery and learning of clinical skills complementing "real" practice. The focus is currently on nurse learning but the principles could equally be applied to multi and inter-professional learning and clinical decision-making. This project represents a new model to enhance clinical skill acquisition and clinical reasoning using a structured competency base. Integral to this is a strong partnership between education and practice utilising "real" live and recorded anonymised patient data from a critical care clinical information system (CIS) within a large district general hospital to structure scenarios fostering problem-based learning. This educational practice interface enables the synthesis of clinical data using virtual technology and sophisticated scenario-based simulation within a skills laboratory. The aim is to enhance the more ad hoc system of learning within conventional practice placements. Early findings suggest that VIP enhances practice providing a safe but challenging learning experience with the benefit of instant performance feedback to students. PMID- 15360913 TI - Mining OMIM for insight into complex diseases. AB - Understanding clinical phenotypes through their corresponding genotypes is one of the principal goals of genetic research. Though achieving this goal is relatively simple with single gene syndromes, more complex diseases often consist of varied clinical phenotypes that may be the result of interactions among multiple genetic loci. Microarray technology has brought the phenotype -genotype relationship to the molecular level, using differently behaving cancers, for example, as the basis for comparing patterns of gene expression. With this feasibility study, we attempted to use similar methods of analysis at the clinical level, in order to evaluate our hypothesis that the clustering of clinical phenotypes would provide information that would be useful in elucidating their underlying genotypes. Because of its breadth of content and detailed descriptions, we used OMIM as our source material for phenotypic and genetic information. After processing the source material, we then performed self-organizing map and hierarchical clustering analysis on representative diseases by phenotypic category. Through pre-determined queries over this analysis, we made two findings of potential clinical significance, one concerning diabetes and another concerning progressive neurologic diseases. Our methods provide a formal approach to analyzing phenotypes among diverse diseases, and may help indicate fruitful areas for further research into their underlying genetic causes. PMID- 15360914 TI - Extracting phenotypic information from the literature via natural language processing. AB - In recent years, the amount of biomedical knowledge has been increasing exponentially. Several Natural Language Processing (NLP) systems have been developed to help researchers extract, encode and organize new information automatically from textual literature or narrative reports. Some of these systems focus on extracting biological entities or molecular interactions while others retrieve and encode clinical information. To exploit gene functions in the post genome era, it is necessary to extract phenotypic information automatically from the literature as well. However, few NLP projects have focused on this. We present the development of a system called BioMedLEE that extracts a broad variety of phenotypic information from the biomedical literature. The system was developed by adapting MedLEE, an existing clinical information extraction NLP engine. A feasibility evaluation study of BioMedLEE was performed using 300 randomly chosen journal titles. Results showed that experts achieved an average precision rate of 65.4%, (95%CI: [58.0%, 72.8%]) and a recall rate of 73.0%, (95%CI: [66.2%, 80.0%]). BioMedLEE had 64.0% precision and 77.1% recall respectively, according to expert agreements. PMID- 15360915 TI - Failure analysis of MetaMap Transfer (MMTx). AB - A pilot study was conducted to evaluate the performance of the MetaMap Transfer (MMTx), a tool that extracts terms from free text and suggests matches to concepts in the Unified Medical Language System' (UMLS'). Five participants, including a content domain expert and a UMLS Expert, manually extracted and mapped terms to UMLS concepts for two disease summary documents from NLM's consumer health site, Genetic Home Reference. The resulting adjudicated annotations were used as a gold standard. Differences in auto-mated term extraction and mapping between MMTx and MetaMap were noted. A failure analysis was conducted to categorize the types of terms not correctly mapped by MMTx. The most frequent type of failure (30%) resulted from missing inferential or world knowledge. Characteristics of each category are discussed. We distinguish between classes of failures that may be easily rectified, such as alternative retrieval strategies to extract exact matches, and ones that require additional research, such as coordinating conjunctions, co-reference resolution, and word sense disambiguation PMID- 15360916 TI - The BioMediator system as a data integration tool to answer diverse biologic queries. AB - We present the BioMediator (www.biomediator.org) system and the process of executing queries on it. The system was designed as a tool for posing queries across semantically and syntactically heterogeneous data particularly in the biological arena. We use examples from researchers at the University of Washington, and the University of Missouri-Columbia, to discuss the BioMediator system architecture, query execution, modifications to the system to support the queries, and summarize our findings and our future directions. Finally, we discuss the system's flexibility and generalized approach and give examples of how the system can be extended for a variety of objectives. PMID- 15360917 TI - Enhancing access to the Bibliome: the TREC Genomics Track. AB - The growing amount of scientific discovery in genomics and related biomedical disciplines has led to a corresponding increase in the amount of on-line data and information. A new challenge for biomedical researchers has been how to access and manage this ever-increasing quantity of information. The Text Retrieval Conference (TREC) has implemented a Genomics Track to create an experimental environment for research in the use of information retrieval systems in the genomics domain. In the first year of the track, an ad hoc document retrieval task and an information extraction task were carried out by 29 research groups. Future work will focus on more complex data sources, searching tasks, and types of experiments. PMID- 15360918 TI - Microarray data mining using gene ontology. AB - DNA microarray technology allows scientists to study the expression of thousands of genes--potentially entire genomes--simultaneously. However the large number of genes, variety of statistical methods employed and the complexity of biologic systems complicate analysis of microarray results. We have developed a web based environment that simplifies the presentation of microarray results by combining microarray results processed for statistical significance with probe set annotation by Genbank, NCBI RefSeqs, GeneCards and the Gene Ontology. This allows rapid examination and classification of microarray experiments--annotated by NCIBI tools --by Statistical Significance and Gene Oncology Classes. By providing a simple, easily understood interface to large microarray data sets, this tool has been particularly useful for small research groups focused on a small number of related genes and for researchers who want to ask simple questions without the overhead of complex data management and analysis. PMID- 15360919 TI - Towards the automatic generation of biomedical sources schema. AB - Biologists and physicians need to access biological and medical data for their experimentations and researches. This information is available on the Internet and is scattered over many heterogeneous data sources. Collecting information is consequently tedious, time consuming and must be improved. To cope with this difficulty, our overall objective is to realize a mediator-based system to integrate heterogeneous biomedical data sources. This requires first an automatic generation of source schema, which is the goal of this work. For that, we describe an algorithm which is based on information extraction. It consists of the extraction of meta-information from each source to infer their schema. Our system enables users to access relevant and specific data, which are up-to-date. To solve the semantic heterogeneity of data sources, we are considering the creation of an ontology. Finally, the management of source evolution is discussed PMID- 15360920 TI - An information extraction and representation system for rapid review of the biomedical literature. AB - With the rapid expansion of scientific research, the ability to effectively find or integrate new domain knowledge in the sciences is proving increasingly difficult. The development of methods and tools for assisting researchers to effectively ex-tract problem-oriented knowledge from heterogeneous and massive information sources, and for using this knowledge in problem-solving is one of the most fundamental research directions for the information and computer sciences today. There is a need for new tools to support more precise identification of relevant research articles and provide visual clues regarding relationships among the document sets. We present the Telemakus system in which aggregated citation information and extracted research findings are displayed in a schema-based document surrogate and an interactive mapping tool provides graphical displays of research inter-relationships from documents across a domain. This system is an innovative approach to creating useful and precise document surrogates and may re-conceptualize the way we currently represent, retrieve, and assimilate research findings from the published literature. PMID- 15360921 TI - A resource to acquire and summarize pharmacogenetics knowledge in the literature. AB - To determine how genetic variations contribute the variations in drug response, we need to know the genes that are related to drugs of interest. But there are no publicly available data-bases of known gene-drug relationships, and it is time consuming to search the literature for this information. We have developed a resource to support the storage, summarization, and dissemination of key gene drug interactions of relevance to pharmacogenetics. Extracting all gene-drug relationships from the literature is a daunting task, so we distributed a tool to acquire this knowledge from the scientific community. We also developed a categorization scheme to classify gene-drug relationships according to the type of pharmacogenetic evidence that supports them. Our resource (http://www.pharmgkb.org/home/project-community.jsp) can be queried by gene or drug, and it summarizes gene-drug relationships, categories of evidence, and supporting literature. This resource is growing, containing entries for 138 genes and 215 drugs of pharmacogenetics significance, and is a core component of PharmGKB, a pharmacogenetics knowledge base (http://www.pharmgkb.org). PMID- 15360922 TI - Clustering gene expression data with temporal abstractions. AB - This paper describes a new technique for clustering short time series coming from gene expression data. The technique is based on the labelling of the time series through temporal trend abstractions and a consequent clustering of the series on the basis of their labels. Clustering is performed at three different levels of aggregation of the original time series, so that the results are organized and visualized as a three-levels hierarchical tree. Results on simulated and on yeast data are shown. The technique appears robust and efficient and the results obtained are easy to be interpreted. PMID- 15360923 TI - Structuring European biomedical informatics to support individualized healthcare: current issues and future trends. AB - Bioinformatics and Medical Informatics are disciplines that up to now have followed separate development with few contacts and synergies between them in Europe. The elucidation of the human genome has however evidenced the need and the possibilities for a strong synergy between the two. Classical epidemiological and clinical research on the one hand, and genomic research on the other, separately considered, are no longer enough for advancing in the so-called genomic medicine, and a new integrative approach is required. Biomedical Informatics is the emerging discipline that aims to put these two worlds together so that the discovery and creation of novel diagnostic and therapeutic methods is fostered. On the basis of the results of the European Commission-funded BIOINFOMED Study, an INFOBIOMED Network of Excellence has been recently constituted with the main objective of setting a durable structure for the described collaborative approach at a European level. Initially formed by fifteen renowned European organisations, the main objective of the INFOBIOMED network is therefore to enable the reinforcement of European BMI as an integrative discipline. PMID- 15360924 TI - Distilling conceptual connections from MeSH co-occurrences. AB - Our aim is to contribute to biomedical text extraction and mining research. In this paper we present exploratory research on the MeSH terms assigned to MEDLINE citations. We analyze MeSH based co-occurrences and identify the interesting ones, i.e., those that are likely to be semantically meaningful. For each selected co-occurring pair we derive a weighted vector representation that emphasizes the verb based functional aspects of the underlying semantics. Preliminary experiments exploring the potential value of these vectors gave us very good results. The larger goal of this project is to contribute to knowledge discovery research by mining the knowledge that is latent within the biomedical literature. It is also to provide a method capable of suggesting cross disciplinary connections via the pairs derived from all of MEDLINE. PMID- 15360925 TI - Methods for multi-category cancer diagnosis from gene expression data: a comprehensive evaluation to inform decision support system development. AB - Cancer diagnosis is a major clinical applications area of gene expression microarray technology. We are seeking to develop a system for cancer diagnostic model creation based on microarray data. In order to equip the system with the optimal combination of data modeling methods, we performed a comprehensive evaluation of several major classification algorithms, gene selection methods, and cross-validation designs using 11 datasets spanning 74 diagnostic categories (41 cancer types and 12 normal tissue types). The Multi-Category Support Vector Machine techniques by Crammer and Singer, Weston and Watkins, and one-versus-rest were found to be the best methods and they outperform other learning algorithms such as K-Nearest Neighbors and Neural Networks often to a remarkable degree. Gene selection techniques are shown to significantly improve classification performance. These results guided the development of a software system that fully automates cancer diagnostic model construction with quality on par with or better than previously published results derived by expert human analysts. PMID- 15360926 TI - Applying task analysis to describe and facilitate bioinformatics tasks. AB - OBJECTIVE: To document bioinformatics tasks currently per-formed by researchers in genomics and proteomics in an effort to recognize unmet informatics needs and challenges, identify system features that would enhance the performance of those tasks, and inform the development of new bioinformatics tools. DESIGN: A cross sectional study of bioinformatics tasks performed by OHSU investigators involved in genomics and proteomics research was conducted using task analysis techniques. RESULTS: Four major categories emerged from 22 bioinformatics tasks reported by 6 research laboratories. These were: 1) gene analysis, 2) protein analysis, 3) biostatistical analysis, and 4) literature searching. Analysis of the data also raised the following challenging issues: 1) lack of procedural documentation, 2) use of home-grown strategies to accomplish goals, 3) individual needs and preferences, and 4) lack of awareness of existing bioinformatics tools. CONCLUSION: Task analysis was effective at documenting bioinformatics tasks performed by researchers in the fields of genomics and proteomics, at identifying potentially desirable system features and useful bioinformatics tools, and at providing a better understanding of some of the unmet needs and challenges faced by these researchers. PMID- 15360927 TI - A log likelihood predictor for genomic classification of oral cancer using principle component analysis for feature selection. AB - DNA microarrays are powerful tools for exploring gene expression and predicting disease state. However, since the number of variables (genes) typically exceeds the number of samples (tissue specimens), many potentially spurious genes may be selected for a predictor function. Principle component analysis (PCA) can greatly reduce the high-dimensional microarray data space while retaining most of the inherent variability. We propose a methodology that uses PCA to identify a predictor vector between two mutually exclusive and collectively exhaustive classes. By projecting the training set upon this vector a distribution of projections can be computed for each class. A log-likelihood ratio is then calculated for class membership. We used this methodology to classify 48 biopsy specimens as either oral squamous cell carcinoma or normal oral mucosa using oligonucleotide microarrays. The system was trained using a set of half the samples, and correctly predicted the membership of the other half. The three most highly positively and three most highly negative predictive genes were all keratins that are known markers of squamous cell carcinoma. PMID- 15360928 TI - Content-based image retrieval for large biomedical image archives. AB - Content-Based Image Retrieval (CBIR) has been a topic of research interest for nearly a decade. Approaches to date use image features for describing content. A survey of the literature shows that progress has been limited to prototype systems that make gross assumptions and approximations. Additionally, research attention has been largely focused on stock image collections. Advances in medical imaging have led to growth in large image collections. At the Lister Hill National Center for Biomedical Communication, an R&D division of the National Library of Medicine, we are conducting research on CBIR for biomedical images. We maintain an archive of over 17,000 digitized x-rays of the cervical and lumbar spine from the second National Health and Nutrition Examination Survey (NHANES II). In addition, we are developing an archive of a large number of digitized 35 mm color slides of the uterine cervix. Our research focuses on developing techniques for hybrid text/image query-retrieval from the survey text and image data. In this paper we present the challenges in developing CBIR of biomedical images and results from our research efforts. PMID- 15360929 TI - The need for a time-machine in the distributed HER. AB - It is widely expected that the Electronic Health Record (EHR) will become an important tool for healthcare professionals when delivering care. The use of this tool will not for long be optional, it will become a professional responsibility to use this tool when appropriate. Until now the focus in the design and implementation of the EHR is on its real-time behaviour. In this paper it is argued that there is a need to be able to reproduce the EHR as it would have presented itself to a specified health care professional at a specified point of time in the past (a "time-machine"). This to be able to assess whether the behaviour of the health care professional was appropriate in view of the data he did retrieve or could have retrieved from the EHR at that point in time. Next the consequences of the implementation of such functionality are explored, these are found to be huge. It would require substantial investments to implement this functionality. So it is important that clarity is created on the need for this time-machine. The professional associations involved and the bodies responsible for the quality of care have to be involved in further discussion on this issue. IMIA might take the lead. Because implementation of the time-machine would have consequences for any information system that contributes in real-time to the EHR, it may well be that the need for this functionality will affect the current common opinion that medical patient data should remain stored in the information systems of the health care establishments where the data were generated and be requested from these systems by the virtual EHR when needed. Uploading medical patient data from the operational systems in the health care establishments to medical data repositories could reduce the number of systems affected considerably. PMID- 15360930 TI - Morpho-functional visualization of dynamic MR-mammography. AB - In view of an increasing use of breast MRI supplementing X-ray mammography, the purpose of this study was the development of a method for fast and efficient analysis of dynamic MR image series of the female breast. The image data sets were acquired with a saturation-recovery-turbo-FLASH sequence facilitating the detection of the kinetics of the contrast agent concentration in the whole breast with a high temporal and spatial resolution. In addition, a morphological 3D FLASH data set was acquired. The dynamic image data sets were analyzed by tracer kinetic modeling in order to describe the physiological processes underlying the contrast enhancement in mathematical terms and thus enable the estimation of functional tissue specific parameters, reflecting the status of microcirculation. To display morphological and functional tissue information simultaneously, a multidimensional real-time visualization system (using 3D-texture mapping) was developed, which enables a practical and intuitive human-computer interface in virtual reality. The spatially differentiated representation of the computed functional tissue parameters superimposed on the anatomical information offers several possibilities: improved discernibility of contrast enhancement, inspection of the data volume in 3D-space and localization of lesions in space and thus fast and more natural recognition of topological coherencies. In a feasibility study, it could be demonstrated that multidimensional visualization of contrast enhancement in virtual reality is practical. Especially, detection and localization of multiple breast lesions may be an important application PMID- 15360931 TI - IRMA--content-based image retrieval in medical applications. AB - The impact of content-based access to medical images is frequently reported but existing systems are designed for only a particular modality or context of diagnosis. Contrarily, our concept of image retrieval in medical applications (IRMA) aims at a general structure for semantic content analysis that is suitable for numerous applications in case-based reasoning or evidence-based medicine. Within IRMA, stepwise processing results in six layers of information modeling (raw data layer, registered data layer, feature layer, scheme layer, object layer, knowledge layer) incorporating medical expert knowledge. At the scheme layer, medical images are represented by a hierarchical structure of ellipses (blobs) describing image regions. Hence, image retrieval transforms to graph matching. The multilayer processing is implemented using a distributed system designed with only three core elements. The central database holds program sources, process-ing schemes, images, features, and blob trees; the scheduler balances distributed computing by addressing daemons running on all connected workstations; and the web server provides graphical user interfaces for data entry and retrieval.. PMID- 15360932 TI - Automatic segmentation of digital micrographs: a survey. AB - Digital micrographs and play a crucial role in today's bio-medical research. Due to progresses in experiment standardization and automation large sets of digital microscopy images, so called micrographs are recorded and stored to databases. The subsequent analysis of the large number of digital images needs the image information to be transformed into quantitative data, which can be processed by statistical methods and datamining. This article summarizes applications of optical microscopy in biomedical research and describes the individual characteristics in micrograph segmentation and classification. An overview on past works based on image processing and artificial neural networks is given and the problem of segmentation evaluation. It concludes with recommendations for future works. PMID- 15360933 TI - Edutainment tools for initial education of type-1 diabetes mellitus: initial diabetes education with fun. AB - PURPOSE: Appropriate initial education for type-1 diabetes mellitus patients is important to prevent late complications. However, type-1 diabetic children have not appreciated traditional learning methods since they rarely contain the elements of fun and interactivity. In this study, we developed, implemented and evaluated a preliminary version of edutainment tools for initial education for type-1 diabetic children. METHODS: Three games running on either personal computer (PC) and GameBoy Advance were developed. All games were designed to educate patients about relationships among food (carbohydrate), plasma glucose level, exercise, and insulin dose. A total of 58 testers evaluated degree of entertainment, usability and clinical usefulness of the games. RESULTS: Generally, testers felt all games were intuitive and fun and the usability of games was highly scored. More than 90% of testers showed an interest in the edutainment approach, and approximately 60% agreed that these games could provide attractive educational environment compared to traditional education, especially for children. CONCLUSIONS: Our edutainment systems were accepted as attractive learning tools for type-1 diabetic children who need initial education. PMID- 15360934 TI - Preparing the next generation of advanced practice nurses. AB - The Institute of Medicine's vision for health professions education highlights the importance of informatics competencies for all graduates. This paper describes an innovative course, required for all graduate students at the University of Colorado Health Sciences Center School of Nursing. The design, implementation, and evaluation of NURS6013, Human Technology Interface provides evidence to support it as one model for consideration in meeting the Institute of Medicine's vision for health professions education: The course provides learning opportunities which examine the impact of technology on society and health care, informatics and human factors, as well as the legal, ethical, and policy issues related to the use of technology. PMID- 15360935 TI - Using Elearning techniques to support problem based learning within a clinical simulation laboratory. AB - This paper details the results of the first phase of a project that used eLearning to support students' learning within a simulated environment. The locus was a purpose built Clinical Simulation Laboratory (CSL) where the School's newly adopted philosophy of Problem Based Learning (PBL) was challenged through lecturers reverting to traditional teaching methods. The solution, a student centred, problem-based approach to the acquisition of clinical skills was developed using learning objects embedded within web pages that substituted for lecturers providing instruction and demonstration. This allowed lecturers to retain their facilitator role, and encouraged students to explore, analyse and make decisions within the safety of a clinical simulation. Learning was enhanced through network communications and reflection on video performances of self and others. Evaluations were positive, students demonstrating increased satisfaction with PBL, improved performance in exams, and increased self-efficacy in the performance of nursing activities. These results indicate that an elearning approach can support PBL in delivering a student centred learning experience. PMID- 15360936 TI - Assessing readability of consumer health information: an exploratory study. AB - Researchers and practitioners frequently use readability formulas to predict the suitability of health-related texts for consumers (e.g., patient instructions, informed consent documents). However, the appropriateness of using readability formulas originally developed for students and educational texts for lay audiences and health-related texts remains to be validated. In this exploratory study, we compared two methods of assessing the readability of consumer health materials: the Cloze procedure, using actual readers, and readability formulas, using our Readability Analyzer program. A statistically significant inverse correlation (r = -0.581, p = 0.01) was found, suggesting that the Readability Analyzer may provide a reasonable "first approximation" for predicting readability of consumer health texts. We also identified several linguistic factors associated with increased reading ease as candidates for improving the performance of the Readability Analyzer. Our ultimate objective is to develop tools to support the design and evaluation of health information that is comprehensible and accessible to laypersons. PMID- 15360937 TI - Virtual Interactive Practice: a strategy to enhance learning and competence in health care students. AB - This paper reports the processes and initial outcomes of a pilot study which investigated a week long 'virtual' children's ward experience for nursing students. Providing sufficient and meaningful experiences which enable students to quickly and effectively achieve competence in diverse areas of practice is often frustrated by the realities of available clinical experiences. Our response to this challenge was to more fully exploit and evaluate technologies which can be used to provide these learning experiences. Students experienced 'real time' scenario based work involving SIM-MAN; interactive information technology scenarios, critical incidents, master classes, video conferencing, and observational skill development exercises. Evaluation methodologies included observation of student performance, competence self rating scales; analysis of videotaped performance episodes and other data generated through the learning activities and lived experience accounts of participants. Initial findings indicate (1) statistically significant improvements in student competence measured through self reports; and (2) evidence of improvement gleaned from observed accounts, video analysis and qualitative evaluative comments. The final outcomes, including work with a control group, will be available for Conference. PMID- 15360938 TI - Design of interactive health drama built on social realism. AB - There are many psychosocial aspects of chronic diseases, such as diabetes. Educational multimedia can support patients with chronic diseases and their families by communicating narratives based on social realism. The production of such socio-realistic interactive health dramas requires systematic methods, especially for the identification of significant stories. The aim of this study is to explore the use of self-documentary video in the design of an Interactive Health Drama environment to support diabetic adolescents and their families. In particular, the potential of the self-documentary video for story development in combination with qualitative interviews were explored. The proposed approach, when further developed, is intended to enable all types of chronic disease patients to work with their specific psychosocial problems in a supportive and stimulating environment adapted to their personality and preferences. PMID- 15360939 TI - The International Partnership in Health Informatics Education. AB - The International Partnership for Health Informatics Education (IPHIE) seeks to promote education through international collaboration of graduate and undergraduate training programs in Medical and Health Informatics. In 1998 an International Partnership of Health Informatics Education was established at six universities: The University of Amsterdam, the Universities of Heidelberg and Heilbronn, the University of Health Informatics and Technology Tyrol at Innsbruck, the University of Minnesota and the University of Utah. The overall goal of this cooperation was to form a network for training and educating medical informatics faculty and students in order to prepare them for leading international positions in medical information and communication technology. In this paper we describe the current state of affairs of IPhiE: the activities undertaken, our experiences, the lessons we have learned over these past five years. In addition we outline our plans for the future. PMID- 15360940 TI - One-hour lecture/demo produces mastery level back safety transfer skills self efficacy in randomized comparisons among hospital nurses. AB - More than 1/3 of back injuries among nurses result from patient transfers. So, a rigorous between-subjects quasi-experimental methodology having broad clinical applicability was developed to evaluate the effects of a one-hour didactic back safety and patient transfer training lecture/demonstration upon 42 recently hired hospital nurses. The design implemented independent random samples including independent control groups, though practical constraints required all 42 to participate in the training. Training effects were assessed for each of three back safety transfer skill self-efficacy measures and their composite total self efficacy score. Four monthly training sessions, each serving different nurses, achieved significant and substantial impacts on both composite total back safety procedure self-efficacy (where effect sizes ranged from approximately 0.8 to 1.8 standard deviations) and on each individual back safety transfer skill self efficacy measure. After training, 85% to 100% of the nurses rated their back safety procedure self-efficacy within three points of the maximum composite total of 30 on the self-efficacy scale. Results may help improve nursing services and identify training strategies to reduce healthcare worker back injuries. PMID- 15360941 TI - Academic performance and comparative effectiveness of computer- and textbook based self-instruction. AB - We intended to clarify the influence of student academic ability on the effectiveness of CAI, using data of a study to assess the effectiveness of a new type of CAI software, cyberPatient (CP), at Kochi Medical School (KMS). A total of 59 third-year students were randomly assigned to four groups: Group-1 used a textbook for self-instruction, Group-2 used CP, Group-3 used both types of learning materials, and Group-4 did not learn. Learning performance was evaluated by multiple-choice examination and OSCE. In order to clarify the influence of students' academic ability on the effectiveness of CAI, statistical analyses were conducted, assigning students as either high or medium or low performance students. High performance students from Group-1, -2 and -3 did not differ significantly in test performance after self-instruction. However, low performance students in Group-1 scored significantly lower than those in Group-2 and -3. All students in Group-2 and -3 reported that CP stimulated willingness to learn and assisted understanding. The present analysis suggested that effectiveness of CAI might be associated with the academic ability of students. PMID- 15360942 TI - International training in health informatics: a Brazilian experience. AB - Technology is transforming not only the practice of health-care but also professional training and educational models. Developing countries, such as Brazil, are increasingly suffering from a severe shortage of health informatics specialists. Training of professionals in this field is expensive, and there is a limited supply of high-quality teaching resources available. We envision that training in health informatics can be better achieved if cultural and technological barriers are anticipated and the training program is prepared accordingly. We describe our four-year experience of a Brazil/USA training program and discuss lessons learned during its implementation. Eleven onsite courses, one seminar, and two conferences were developed under this unique initiative, which made possible the collaboration among different countries and distinguished leaders in the field of medical informatics. PMID- 15360943 TI - Preparing student nurses, faculty and clinicians for 21st century informatics practice: findings from a national survey of nursing education programs in the United States. AB - Because healthcare delivery increasingly mandates data-driven decision-making, it is imperative that informatics knowledge and skills are integrated into nursing education curricula for all future nurse clinicians and educators. A national online survey of deans/directors of 266 baccalaureate and higher nursing education programs in the U.S. identified perceived informatics competencies and knowledge of under-graduate and graduate nursing students; determined the preparedness of nurse faculty to teach and use informatics tools; and elicited perceptions of informatics requirements of local practicing nurses. Frequency data and qualitative responses were analyzed. Approximately half of the programs reported requiring word processing and email skills upon entry into the nursing major. The use of standardized languages and the nurse's role in the life cycle of an information system were the least visible informatics content at all levels. Half of program faculty, rated as "novice" or "advanced beginners", are teaching information literacy skills. Findings have major implications for nurse educators, staff developers, and program administrators who are planning faculty/staff development opportunities and designing nursing education curricula that prepare nurses for professional practice. PMID- 15360944 TI - Registration and regulation of health informatics professionals in the UK National Health Service. AB - Whilst good health informatics can contribute directly to the delivery of effective patient care, bad informatics can kill. The UK is establishing a regulatory body to register health informatics specialists who can demonstrate that they meet agreed standards of professional practice: the UK Council for Health Informatics Professions (UKCHIP). UKCHIP will also manage the introduction of a Code of Conduct (based on the IMIA Code) and processes for continuous professional development, appeals, and removal from the Register. The challenge has been to design a process that provides a suitable structure for all informatics staff, including records, coding, audit, library and knowledge management, ICT, systems, information management and clinical management specialists. The contention is that this inclusive model of regulation has international applicability. PMID- 15360945 TI - Assessing informatics in Canadian schools of nursing. AB - The provision of informatics content in the basic education programs for Canadian nurses has been limited to date. In previous years, efforts to engage nurse educators in discussions regarding the significance of informatics for tomorrow's nurses had been met with limited interest and understanding. There was an obvious need to heighten the awareness and active participation of nurse leaders in the development of strategies to attend to the informatics education needs of Canadian nurses. This paper describes the findings of a national study to understand the current state of informatics integration within basic nursing curricula PMID- 15360946 TI - Developing an interaction-centered evaluation tool for distance education. AB - Web-based distance instruction is growing in popularity. As more courses and programs go online, instructional methods and technologies are changing to meet new demands. These courses and instructional methods need be evaluated to determine their quality and to aid in their development. An instrument that measures the quality of student interaction within courses could play a useful role in this evaluation. The objective of this project was to produce an interaction-centered evaluation tool for distance education. We developed items for the evaluation tool based on a review of the literature and interviews with distance education faculty. We then conducted a measurement study to determine the instrument's validity and reliability. Fifty-five students in seven medical informatics courses participated in this study. Questionnaire items were included or rejected based on their fit with underlying theoretical constructs. In conclusion, the Web-based evaluation tool developed in this project measures student perception of quality of course interactions with demonstrated validity and reliability. PMID- 15360947 TI - CAMPUS--a flexible, interactive system for web-based, problem-based learning in health care. AB - Care for patients is the best way to learn medicine and medical methods and skills. But the availability of real patients for learners is limited. Often the appropriate patient is missed or he can not be demonstrated to all students because of practical or ethical problems. A possible solution is the presentation of a clinical problem by a virtual patient using a simulative computer program. But such an approach means often a lot of work for the author of the case. We developed a flexible and realistic system, CAMPUS (www.medicase.de), which supports the case authors with appropriate vocabularies and a comfortable authoring tool. There are different kinds of case presentations to the users in accordance to the level of professionalism of the user and the scenario the program is used in. An evaluation within an internship has shown, that such pro grams are judged as useful by medical students. But further studies are necessary to examine if the program will be used for the self-study and to develop further kinds of using scenarios within problem-based curricula. PMID- 15360948 TI - Improving governance and reducing risk in electronic patient record systems: ensuring appropriate competencies for support and end-user staff. AB - As clinical health information systems, particularly Electronic Patient Records, become widely used, so it is important to address the Governance and Risk Reduction policies needed to ensure their safe and effective use. Greater emphasis needs to be placed on ensuring that all staff involved have relevant, recognized competencies. This paper describes two initiatives--one for health informatics staff, and one for end-user health professionals--ensure and assess such competencies PMID- 15360949 TI - Helping healthcare consumers understand: an "interpretive layer" for finding and making sense of medical information. AB - Healthcare consumers need to find, comprehend, and interpret health information before making informed decisions. Recent work by others and our own work suggest that mis-matches in representations of health information used by consumers and professionals occur at different levels of knowledge representation, such as terminology (i.e., form or surface structure and concept or meaning) and semantic relation-ships. A challenge for consumer health informatics research is to devise a comprehensive strategy to bridge the gap between consumer understanding and biomedical knowledge at all levels. We propose a framework to inform the design of an "interpretive layer" to "mediate" between lay (illness model) and professional (disease model) perspectives, at all levels. In our view, the goal is to assist consumers in identifying terms to describe their needs, finding and understanding relevant information, and applying that knowledge for informed healthcare decision making. PMID- 15360950 TI - Readiness for evidence-based practice: information literacy needs of nurses in the United States. AB - In this paper U.S. nurses' readiness to provide Evidence-Based Practice (EBP) as measured by their information literacy knowledge and skills is described. The Institute of Medicine directed health care providers to use EBP as a means to improve patient safety, efficiency and effectiveness of health care services. Information literacy has been identified as a nursing informatics competency for the basic nurse. As such, information literacy is an essential component in the application of EBP. The importance of developing information literacy skills is enhancement of the nurse's ability to use current best available research literature in the conduct of EBP with subsequent improvement in nursing sensitive patient outcomes. This study describes the level of nurses' information literacy knowledge and gaps in their skills for identifying, accessing, retrieving, evaluating and utilizing research evidence to provide best care for patients. The value of this study is to increase awareness among nurse administrators, educators, and clinicians of the need for information literacy education to enable evidence-based nursing practice and to guide development of supportive curricula and professional continuing education. PMID- 15360951 TI - Towards integration of computer games in interactive health education environments: understanding gameplay challenge, narrative and spectacle. AB - CONTEXT: There is an alarming progress in the health status of the young in western countries, and new methods and tools for behavioural health interventions are urgently called for. OBJECTIVE: To explore how computer game designs can be integrated in the development of Interactive Health Education Environments. DESIGN: Qualitative analyses of adolescents' experiences of playing an action adventure computer game, using data from in-depth interviews. RESULTS: A model is presented, where the gameplaying experience is connected to four components of computer games. Playing computer games was found to mainly be motivated by the challenges and competition represented in the gameplay scripts. CONCLUSIONS: Interactive health education environments can be improved by implementing challenging gameplay scripts, spectacular technical features and narratives. PMID- 15360952 TI - Experiments in cross-language medical information retrieval using a mixing translation module. AB - Given the ever-increasing scale and diversity of medical literature widely published in English on the Internet, improving the performance of information retrieval by cross-language is an urgent research objective. Cross-language medical information retrieval (CLMIR) consists of providing a query in one language and searching medical document collections in one or more different languages. Our users of CLMIR are users who are able to read biomedical texts in English, but have difficulty formulating English queries. This paper proposes a French/English CLMIR system as a mixing model for supporting the retrieval of English medical documents. Methods fall into the category of query translation approach in which we use a hybrid machine translation that combines a pattern based module with a rule-based translator and includes three steps from pre- to- post-translation. In parallel to this hybrid machine translation, we use multilingual UMLS Methasaurus as a complementary translator. The results show that using a mixing translation module outperforms machine translation-based method and thesaurus-based method used separately. PMID- 15360953 TI - Telemedicine in Russia. AB - There are quite a number of the Russian medical institutions using different telemedicine technologies. There are also several telehealth networks created around the scientific medical centers and large hospitals. They use store-and forward technology and real-time video conferencing over ISDN and IP-channels. Quality issues, management and legal issues, integration and financing issues, future development of the Russian telehealth networks are discussed. PMID- 15360954 TI - Computer-based and live interviews on problem drinking: users' attitudes. AB - Some studies of computerized interviews particularly those that deal with personally sensitive topics demonstrate that people have a preference for automated interviews versus live interviews. To explore this phenomenon, we administered four open-ended questions after participants were screened for problem drinking by both an automated and a human telephone interviewer. Both interviews administered AUDIT (Alcohol Use Disorders Identification Test) for assessing problem drinking. Individuals were recruited into the study who responded to ads in daily papers. Sixty-two percent of the participants preferred the human interviewer and only 3% among these expressed a concern about confidentiality of the interview. Among the 22% who preferred the automated interview, 32% indicated confidentiality as a reason for their preference. PMID- 15360955 TI - Towards a virtual health record for mobile home care of elderly citizens. AB - Mobile work situations within home care of the elderly require immediate and ubiquitous access to patient-oriented data. The ongoing Swedish research project "Technical support for Mobile CloseCare" focuses on the development and evaluation of work-scenario oriented ICT support for enhanced home care of elderly citizens. The aim of the project is to provide a seamless and consistent information flow between different health care providers and to give intuitive access to information services for the elderly and their relatives. For that purpose, different independent software components are connected through a mobile communication platform. Flexible access to prioritized information for different users in different work situations will be given through a virtual health record. In order to obtain both usable and clinically relevant results, a user centered system development approach is followed. Evaluation of the project results will be based on usability tests and quasi-experimental studies on how system implementation influences quality of care and job- and life satisfaction for care providers, patients and relatives. PMID- 15360956 TI - Communication and information needs and barriers: an international collaboration model. AB - This future vision paper describes the emerging issue of multicultural medical informatics teams. In an increasingly networked healthcare environment, the rise of global teams has come to the forefront, calling for an increased understanding of various cultural norms. Team members may reside in other countries and be known to others only virtually. Concomitantly, the team dynamics of physically present members may be impacted by international factors such as culture of origin. PMID- 15360957 TI - The medical appointment scheduler. AB - In order to enhance our understanding of how to best improve patient care, it was necessary to initially identify key questions that patients ask most frequently by recording actual calls made by home hemodialysis patients to a dialysis clinic over a three-month period. From an initial analysis of the recorded conversations, one of the most frequent reasons for patient calls was for scheduling concerns. Since the use of automated systems often enable a quicker response to patients' needs and alleviate time demands on the personnel that have to answer these calls, an automated scheduling system was designed and implemented to address this concern. The SCHEDULER is a mixed-initiative spoken dialogue system that allows a patient to schedule an appointment with a provider over the telephone. The system was implemented using SPEECHBUILDER, a utility developed by the Spoken Language System group at M.I.T. that helps automatically configure human language technology servers to create a new conversational system. We describe the system architecture, general considerations in the design, and a preliminary evaluation of the system. PMID- 15360958 TI - Cognitive human factors for telemedicine systems. AB - The recent integration of telephony systems with information and communication technology (ICT) enables the development of innovative tools for telemedicine. The dissemination and widespread acceptance of telephone-based care monitoring systems challenge the researcher to deal with the cognitive factors involved in the patient-physician interaction, and the way they should be to shape up the technological solutions. This paper proposes a model that describes the impact of socio-cognitive factors in the complex process of health care management. The model has been used to design and develop a telephone system for the management of hypertensive patient within the EU funded Homey project. The knowledge existed in a widely accepted guideline for the care of hypertension has been represented and augmented through the proposed cognitive model. The final product is an intelligent system able to manage an adaptive dialogue. It monitors patients' adherence and increases their involvement by promoting self-care through frequent virtual visits, which is complementary to the traditional face-to-face encounters with their primary care physicians. PMID- 15360959 TI - Approaches towards a regional, shared electronic patient record for health care facilities of different health care organizations--IT-strategy and first results. AB - Today, information processing in healthcare facilities is usually primarily directed towards the information needs of the respective institution. This stands in apparent contradiction to the fact that patients may not be solely treated in one general practice or hospital. More information processing towards patient centered, shared care would better support high quality as well as efficient health care. We developed a stepwise approach transforming trans-institutional information system architectures (TISAs) from an inefficient state caused by redundancy and media cracks towards a state which better supports patient centered, shared care. In a total of three steps we want to establish electronic communication between existing information systems of different healthcare facilities for transmission of discharge summaries and diagnostic results. In further stages we plan to expand this communication solution to a regional comprehensive and consistent electronic patient record for multiple hospitals and general practices in Tyrol, Austria. In addition, two related approaches towards supporting shared care have been compared with our approach. The aim of this paper is to report on our approach and first experiences. PMID- 15360960 TI - The impact of ICT on communication in healthcare. AB - Communication processes are pervasive in the daily practice of health professionals. Reorganizing these daily practices by introducing ICT, inevitably effects one or more communication processes. Understanding exactly what these effects are, is a major problem in designing and implementing ICT-applications. In this paper we present an analysis of these effects, based on a theory of communication processes. The concept of 'decoupling' is pivotal in our analysis. Based on the identified effects, and some preconditions that have to be met in order for these effects to take place, we derive a number of guidelines for reorganizing communication processes by means of applying ICT. The application of these guidelines will be demonstrated and discussed in the context of the reorganization of a team conference at the Rehabilitation Clinic of the Rheumatology Department of the Leiden University Medical Center (RCRD/LUMC). PMID- 15360962 TI - A tale of two hospitals: a sociotechnical appraisal of the introduction of computerized physician order entry in two Dutch hospitals. AB - We compared the implementation of computerized physician order entry (CPOE) in two Dutch hospitals, one being an academic medical center and the other a large regional non-academic hospital. Both implemented the TDS7000 system that was running on the same computer, located in the computing department of the academic medical center. The outcomes of the implementation were different. The introduction of CPOE in the university medical center failed, while it was a success in the non-academic hospital. An appraisal of the different outcomes is possible when we consider the implementation of information as a thorough social process in which the technical and the social are closely interrelated. Our findings suggest that organizational change associated with CPOE implementation should not focus on individual physician behavior but on medical work as a collaborative professional effort PMID- 15360961 TI - An information flow analysis of a distributed information system for space medical support. AB - In this study, we applied the methodology grounded in human-centered distributed cognition principles to the information flow analysis of a highly intensive, distributed and complex environment--the Biomedical Engineer (BME) console system at NASA Johnson Space Center. This system contains disparate human and artificial agents and artifacts. Users and tasks of this system were analyzed. An ethnographic study and a detailed communication pattern analysis were conducted to gain deeper insight and better understanding of the information flow patterns and the organizational memory of the current BME console system. From this study, we identified some major problems and offered recommendations to improve the efficiency and effectiveness of this system. We believe that this analysis methodology can be used in other distributed information systems, such as a healthcare environment. PMID- 15360963 TI - An intelligent case-adjustment algorithm for the automated design of population based quality auditing protocols. AB - We develop a method and algorithm for deciding the optimal approach to creating quality-auditing protocols for guideline-based clinical performance measures. An important element of the audit protocol design problem is deciding which guide line elements to audit. Specifically, the problem is how and when to aggregate individual patient case-specific guideline elements into population-based quality measures. The key statistical issue involved is the trade-off between increased reliability with more general population-based quality measures versus increased validity from individually case-adjusted but more restricted measures done at a greater audit cost. Our intelligent algorithm for auditing protocol design is based on hierarchically modeling incrementally case-adjusted quality constraints. We select quality constraints to measure using an optimization criterion based on statistical generalizability coefficients. We present results of the approach from a deployed decision support system for a hypertension guideline. PMID- 15360964 TI - A management information system model for process-oriented health care. AB - OBJECTIVE: To develop a conceptual model of a management information system for process-oriented health care organizations. METHODS: Qualitative data was collected from two case studies in process-oriented health care settings. The first study addressed the information requirements of health care managers and the second study focused on organizational activities and clinical practice. From these data, preliminary models were iteratively developed, interpreted, and further revised. SETTING: A county hospital in southern Sweden with 30 clinics and 3,200 employees. RESULTS: A conceptual model of a management information system for process-oriented health care organizations was developed in two parts: one part that describes the organizational interface of the model and the other part that describes the architecture of the model. CONCLUSION: A conceptual model has been developed for local-level integration of management information systems and organizational procedures in process-oriented health care organizations PMID- 15360965 TI - Adding insight: a qualitative cross-site study of physician order entry. AB - The research questions, strategies, and results of a six-year qualitative study of computerized physician order entry implementation (CPOE) at successful sites are reviewed over time. The iterative nature of qualitative inquiry stimulates a consecutive stream of research foci which, with each iteration, add further insight into the overarching research question. A multidisciplinary team of researchers studied CPOE implementation in four organizations using a multi method approach to address the question "what are the success factors for implementing CPOE?" Four major themes emerged after studying three sites; ten themes resulted from blending the first results with those from a fourth site; and twelve principles were generated when results of a qualitative analysis of consensus conference transcripts were combined with the field data. The study has produced detailed descriptions of factors related to CPOE success and insight into the implementation process. PMID- 15360966 TI - A usability study of CPOE's medication administration functions: impact on physician-nurse cooperation. AB - Implementation of CPOE systems in Healthcare Institutions has proven efficient in reducing medication errors but it also induces hidden side-effects on Doctor Nurse cooperation. We propose a usability engineering approach to this problem. An extensive activity analysis of the medication ordering and administration process was performed in several departments of 3 different hospitals. Two of these hospitals are still using paper-based orders, while the 3rd one is in the roll-out phase of medication functions of its CPOE system. We performed a usability assessment of this CPOE system. The usability assessment uncovered usability problems for the entry of medication administration time scheduling by the physician and revealed that the information can be ambiguous for the nurse. The comparison of cooperation models in both situation shows that users tend to adopt a distributed decision making paradigm in the paper-based situation, while the CPOE system supports a centralized decision making process. This analysis can support recommendation for the re-engineering of the Human-Computer Interface. PMID- 15360967 TI - Q.U.i.P.S.--a quality model for investigating risk exposure in e-health systems. AB - The increasing dependency being placed on electronic health information systems presents new challenges for today's health managers and systems developers. This paper uses Electronic Health Records as an example to demonstrate how there are four critical attributes for e-health system development. To produce dependable and viable IT solutions, each attribute needs to be specifically addressed and prioritized. It will be shown how these attributes possess a number of interdependencies making the analysis and prioritization tasks complex and hence, in practice, often incomplete. The proposed QUiPS model aims to provide a complete framework for building trustworthy solutions. It identifies the pertinent issues and the tools and techniques needed to determine the risk exposure with a given system. The results from one of the Case Studies that focuses on Usability is detailed, while other studies relating to Safety and Privacy are outlined. The outcomes to the proposed approach are e-health systems that the clinicians and patients alike can trust and will, therefore, be fully accepted. PMID- 15360968 TI - Integration of electronic patient record context with message context. AB - A methodology to construct specific messages with clear objectives inside clinical processes, while simultaneously including contextual information, remains a problem today. This paper addresses the issue of combining specific message context (process driven) with the context of a patient record (patient centered). In Belgium, simplified conceptual models for Electronic Patient Record (EPR) architecture and for message architecture, based on previous comprehensive and international work, have been produced, validated and mapped into an integrated message format. The resulting model described in this paper highlights the main conceptual links between both basic models: at the action level and at the Transaction level. Using XML, some parts of the model have already been implemented in various national projects. Key lessons learned may be imported at the international level. PMID- 15360969 TI - Experiences with the development, implementation and evaluation of automated decision support systems. AB - A framework for the implementation of guidelines--GASTON--was developed. Its functions range from the definition of guidelines (using a GLIF like approach) to the implementation of a DSS that can be coupled to existing information systems. This paper discusses experiences with the development of three systems in different domains. It is concluded that the toolbox corresponding to the framework could be successfully used to develop these systems. PMID- 15360970 TI - User acceptance of clinical information systems: a methodological approach to identify the key dimensions allowing a reliable evaluation framework. AB - The introduction of Computerized Information Systems (CIS) in clinical settings encountered difficulties. These difficulties highlight the lack of understanding of factors and mechanisms influencing user acceptance. The existing tools and research obtained contradictory results that point out the existence of neglected aspects, such as impacts of CIS, in computer science developments in complex settings. This paper proposes to identify key dimensions which make up user acceptance in clinical settings through the union of three methods. They define five main dimensions which require a concrete evaluation to validate the underlying proposed framework and to complete the description of the acceptance phenomenon. Identifying key dimensions opens the gate to comparative evaluation of many CIS and adds new indicators to evaluate the highlighted dimensions. A long-term aim is the development of longitudinal studies and to state priorities and guidelines for new CIS designs. PMID- 15360971 TI - Is it possible to collect medicalized data in Africa? An experience in the pediatrics service in the University Hospital in Yopougon, Abidjan, Ivory Coast. AB - The aim of this study is to demonstrate that it can be relevant to implement medicalized data in the African hospitals in order to establish morbidity statistics and economical evaluation. A retrospective survey has been carried out on 300 children hospitalized in the paediatrics service in the Yopougon University Hospital from January 1st to December 31st 1999. Data have been collected on forms similar to the ones used in France for discharge summaries. The file batching used for these 300 files is the French DRG named GHM: it shows that, once the files have been divided up in main categories(CMD), CMD 18 (infectious and parasitic diseases) amounts to nearly half of the stays (43.7%). Likewise, in the Case Mix, GHM 611 (others infectious or parasitic diseases) represents, with 38%, more than one third of the stays. The most frequent diagnosis is the B50.9, plasmodium falciparum malaria without precision, with 24.3% of the stays. The patients' stays are shorter than in France. But according to the quality of the collected data, the inadequacy of the tools used and the category of the patients taken into consideration, these results have to be interpreted with restriction. However, this survey shows that it is possible to collect medicalized data in the African hospitals. Other services should feel concerned and more appropriate classifications that would reckon with epidemiological specificities should be applied. PMID- 15360972 TI - Design of a patient-centered, multi-institutional healthcare information network using peer-to-peer communication in a highly distributed architecture. AB - OBJECTIVE: to design a community healthcare information network for all 450,000 citizen in the State of Geneva, Switzerland, connecting public and private healthcare professionals. Requirements include the decentralized storage of information at the source of its production, the creation of a virtual patient record at the time of the consultation, the control by the patient of the access rights to the information, and the interoperability with other similar networks at the national and european level. METHODS: a participative approach and real world pilot projects are used to design, test and validate key components of the network, including its technical architecture and the strategy for the management of access rights by the patients. RESULTS: a distributed architecture using peer to-peer communication of information mediators can implement the various requirements while limiting to an absolute minimum the amount of centralized information. Access control can be managed by the patient with the help of a medical information mediator, the physician of trust. PMID- 15360973 TI - Integration challenges of clinical information systems developed without a shared data dictionary. AB - Legacy systems have proven to be long-term integration challenges for Intermountain Health Care (IHC) despite commitment and attention to share clinical information across settings and among clinicians. This study measures the extent of the disparity of data elements across three independent data systems in current use. A sample of relevant data elements was selected across systems covering prenatal, labor and delivery, and newborn intensive care units (NICU). The findings revealed only 17% of these sample data elements had compatible structure across all three systems. The implications from differences in granularity, missing data, and duplicate data entry, include diminished data quality, greater risk for medical error, increased costs of integration and inefficient use of clinician time. Retrospective guidelines for managing conceptual context and granularity are given to assist in designing an integrated longitudinal patient electronic medical record. PMID- 15360974 TI - Integrating detailed clinical models into application development tools. AB - Several groups are currently working on defining detailed clinical models (also called templates or archetypes) that are refinements of abstract medical models like the HL7 (Health Level Seven) Reference Information Model. At IHC, we have created over 3,000 detailed clinical models in the last five years. These models have become an essential part of the architecture of our electronic medical record (EMR) system. As a result, we have created an increasingly sophisticated set of tools that allow the models to be searched, viewed, and ultimately incorporated into medical applications. These browsers have some commonality with terminology browsers, but are distinct in that the explicit structure of the information models must be accommodated. In this paper we report our experience in making browsers for detailed clinical models that are integrated with application authoring tools. PMID- 15360975 TI - Computer-based drug ordering: evaluation of interaction with a decision-support system. AB - Provider order entry systems (POE) often incorporate active decision-support component for drug dosing. The efficacy of automated alerts that suggest dose amounts to the clinician in real time depends in part on how well they are timed to fit into the decision process and on their representational structure. We have conducted a cognitive evaluation of an interaction with a POE system that offered active decision support for heparin dosing with the goal of characterizing its effectiveness and opportunities for error. Two researchers completed a cognitive walk-through of an ordering task based on a clinical scenario. In addition, seven clinicians were asked to enter a set of orders in an experiment using the same scenario. The analysis revealed that users without a solid conceptual knowledge of the ordering system followed patterns of inefficient interactive behavior resulting in delays and some errors. Physicians often did not take full advantage of automatic dose computation provided by a decision support component and used it largely as reference. The calculated dose was not perceptually salient in the generated alert and required users to engage in meaning interpretation of the displayed information. Better visual presentation of the alert message would likely result in faster and less cognitively demanding interaction. PMID- 15360976 TI - UMIN--current status and future perspectives. AB - UMIN, fully funded by the Japanese government, is the largest and most versatile public academic information center for biomedical sciences in the world, and is considered as indispensable information infrastructure for the Japanese medical community. As of April 1st 2004, all Japanese national universities, including UMIN, will be incorporated as independent administrative entities. After the incorporation, each national university will have authority to determine its own budget, organization, etc. by itself, and will also be able to carry out profit making business related to its activities under certain constraints. Thus we suspect that UMIN will inevitably be faced with a strong demand to undertake profit-making business at an early stage. Even in this case, we believe that UMIN should continue to provide its information services to the Japanese medical community free of charge as it has done, utilizing earnings from its profit making business that services commercial companies, because the success of UMIN has depended on the cooperation and sympathy of the community in the past, and will continue to do so. PMID- 15360977 TI - The relationship of usability to medical error: an evaluation of errors associated with usability problems in the use of a handheld application for prescribing medications. AB - This paper describes an innovative approach to the evaluation of a handheld prescription writing application. Participants (10 physicians) were asked to perform a series of tasks involving entering prescriptions into the application from a medication list. The study procedure involved the collection of data consisting of transcripts of the subjects who were asked to "think aloud" while interacting with the prescription writing program to enter medications. All user interactions with the device were video and audio recorded. Analysis of the protocols was conducted in two phases: (1) usability problems were identified from coding of the transcripts and video data (2) actual errors in entering prescription data were also identified. The results indicated that there were a variety of usability problems, with most related to issues of ease of use. In addition, other problems were identified which were related to limitations of the content of the program. In examining the relationship between usability problems and errors, it was found that certain types of usability problems were closely associated with the occurrence of specific types of errors in prescription of medications. Implications for the improvement of safety of health care information systems are discussed. PMID- 15360978 TI - The Success Factor Profile for clinical computer innovation. AB - Fifty to seventy percent of information system projects fail. Most of the failures are not the victims of flawed technology, but rather organizational and people related issues. When Vanderbilt University Medical Center began an intensive electronic health record (EHR) effort, a process was carefully designed to select the clinical areas where new tools could be developed and pilot tested. The Success Factor Profile was created to guide the selection of sites most likely to have innovation success. This paper describes both the tools and the processes used to select clinical sites for new computer tools development and pilot implementation. Early results demonstrated that the tools provided structure for the decision making process, permitting side-by-side comparison of "apples and oranges." Selecting the site most likely to succeed with computer application innovation and early implementation has broad applicability in healthcare informatics. Failure to succeed with early system users is not only costly, but also discourages users and developers alike, and may damage the reputation of the tools and systems across the institution. PMID- 15360979 TI - Managing the tensions between national standardization vs. regional localization of clinical content and templates. AB - The deployment of sophisticated software tools and electronic health records offers many new opportunities and challenges to support care delivery. One of the key opportunities is to enhance the quality of care with evidence-based medicine (EBM). One of the key challenges is to embed EBM in tools that directly facilitate the process of documentation and care delivery. Since clinicians typically have the option of using free text for most of their documentation, the tools that provide embedded EBM must be at least as efficient as free text. There are many requirements that must be met in order to effectively embed EBM within clinical content tools and enhance both the usability and the actual use of such tools and clinical content: (1) Facilitate the documentation process; (2) Facilitate the care delivery process, e.g. make order entry faster; (3) Contain recommendations that are highly relevant to the clinical context of an encounter; (4) Aid in the capture of discrete coded data. Support for local variation is often key to meeting these objectives and becomes a central factor in helping clinicians shift from unstructured free text, to the use of these tools, which support the delivery of EBM. This document describes the central tension between the objective of national standardization and delivery of EBM and the need for regional localization of clinical content. This tension must be thoughtfully managed to maximize the quality of care delivery and associated workflow practices. The key elements of legitimate local variation that must be recognized in order to achieve these goals are described in this document, and the key principles for managing the tensions between generalization and localization are identified. PMID- 15360980 TI - Seamless information systems for critical care--myth or magic? AB - A multi-site study is described that evaluates the scope for seamless Clinical Information Systems (CIS) in critical care. Observations, shadowing of clinical staff, interviews and questionnaires show a triangulation in data collection methods as well as location triangulation as the study is conducted across four sites, two each in UK and Denmark. Role Activity Diagrams (RAD) are used to capture critical care work processes. The RADs are analysed to show the 'simplified complexity' of the work processes, which are changed by the introduction of information systems for the everyday use and management of all clinical information. Further, CIS that reconcile expectations of both hospital management and clinical staff and that have the potential to adapt to their organisational environment have a greater chance of surviving in autopoietic organisations such as critical care. Despite decades of Informatics, no such system exists in its entirety; this study shows that 'ancient problems' of clinical information systems development, implementation and integration are still heavily prevalent. However, the potential for CIS that consider integrating all clinical information requirements is immense. PMID- 15360981 TI - A novel diagnostic aid (ISABEL): development and preliminary evaluation of clinical performance. AB - Clinical diagnostic aids are relatively scarce, and are seldom used in routine clinical practice, even though the burden of diagnostic error may have serious adverse consequences. This may be due to difficulties in creating, maintaining and even using such expert systems. The current article describes a novel approach to the problem, where established medical content is used as the knowledge base for a pediatric diagnostic reminder tool called ISABEL. The inference engine utilizes advanced textual pattern-recognition algorithms to extract key concepts from textual description of diagnoses, and generates a list of diagnostic suggestions in response to clinical features entered in free text. Development was an iterative process, relying on sequential evaluation of clinical performance to provide the basis for improvement. The usage of the system over the past 2 years, as well as results of preliminary clinical performance evaluation are presented. These results are encouraging. The ISABEL model may be extended to cover other domains, including adult medicine. PMID- 15360982 TI - Outcome measures in clinical information systems evaluation. AB - Systems developed through informatics methods can be evaluated at different levels, depending on the purpose of the evaluation. A key class of measures useful in relating information systems to the quality of patient care is benefit or out-come measures, which reflect differences in the health or welfare of the patient that result from the system being utilized. A framework for describing evaluation measures is presented, definitions are discussed, and the results of a literature review are presented to indicate outcome measures used in medical informatics over several decades. The conclusion suggests that more attention should be given to the standardization of terminology and to outcome measures and methods. PMID- 15360983 TI - Reasons for physician non-adherence to electronic drug alerts. AB - CONTEXT: Many adverse drug errors may be prevented through electronic order entry systems that provide decision support to physicians by screening prescriptions for dosing errors, drug-disease, drug-allergy and drug-drug interactions. The adherence to such decision aids is varied and the reasons for this variance not well understood. OBJECTIVE: To assess the feasibility and performance auto-mated drug alerts within an electronic decision support system for physician prescribing. METHODS: Drug alert data were collected from a pilot project with 30 participating general practitioners who were provided with interactive electronic prescription capabilities through a personal digital assistant (PDA). RESULTS: 66,642 electronic prescriptions resulted in a total of 1,869 drug alerts. The most common alert types were analysed, along with reasons for non-adherence to automated drug alerts. CONCLUSIONS: Non-adherence to alert information appears to be associated with additional knowledge of the clinical situation, beyond that inherent in the decision support tool, for the specific patient context. Further work is required to understand how best to provide this type of support to physicians. PMID- 15360984 TI - A new quality improvement study every day? Using QTools to build quality improvement projects around primary care electronic medical record systems. AB - Greater and more appropriate use of information technology has been proposed by the US Institute of Medicine as one way in which the quality of health care can be improved. Electronic medical record systems offer particular advantages because they can support clinicians at the point of care as well as allowing patient care to be monitored over time. Developing quality improvement tools that make use of the electronic medical record can, however, be difficult and often requires special software. This paper describes a system called QTools that provides a range of general-purpose tools to support quality improvement work in Norwegian primary care. The paper discusses how QTools has been used in two studies involving over 100,000 patient consultations and how it is supporting current work to improve the prescription of anti-hypertensive drugs. Features that have not yet been used in a quality improvement study are also discussed. PMID- 15360985 TI - Archetypes: the PropeR way. AB - The PropeR project studies the effect of Decision Support in an Electronic Health Record system (EHR) on the quality of care. One of the applications supports a multidisciplinary primary care team rehabilitating stroke patients in their home environment. This project required an EHR system that could handle information of multiple disciplines and multiple, distributed data sources. It should also be flexible enough to handle an entirely different domain with only minor modifications. The resulting EHR system is a distributed system based on international standards and reusable components. It is generic in nature since all references to the domain are stored in separate XML documents: the clinical data are defined through a modified version of the OpenEHR archetypes, described in XML and views on the data, either for review or for data entry are also described in XML documents. Currently, the system is being evaluated by a care team using laptops with a wireless internet connection. The use of standards greatly improves the quality and reusability of the resulting software system, but they do not solve the issues that surface during implementation, such as context and screen representation. PMID- 15360986 TI - An adoption study of a clinical reminder system in ambulatory care using a developmental trajectory approach. AB - In this study, we assess 41 medical residents' acceptance and adoption of a clinical reminder system for chronic disease and preventive care management in the ambulatory care environment using a novel developmental trajectory approach. This group-based, semi-parametric statistical modeling method identifies distinct groups, following distinct usage trajectories, among those who recorded use of the reminder system within an evaluation period of 10 months. We trace system use within these groups over time using computer-generated logs and user satisfaction surveys. Our preliminary analysis of this small sample of users delineates three categories of users. Feedback from these categories of users is being used to re engineer the application and adapt it better to their workflow and functionality requirements. Despite the small sample size in this particular study, we conclude that this methodology has considerable promise to provide new insights into system usability and adoption issues that may benefit clinical decision support systems as well as information systems more generally. PMID- 15360987 TI - Constructing and validating a consumer health portal rating index. AB - Web-based consumer health information portals are on the increase. With such a large number of health related sites available, design and information quality are key concerns. This article provides an evaluation framework based on the work of Slack (2001) and the HONcode guidelines. Research shows that greater transparency is needed with regard to how rating instruments are constructed and validated. The consumer health portal index developed in this research has been tested for face and content validity through a series of focus groups following Sullivan's 5 step process. PMID- 15360988 TI - Evaluation of consumer health website accessibility by users with sensory and physical disabilities. AB - Growth of the World Wide Web is beginning to create new opportunities for direct patient access to health care resources. At the same time, advances in medical care have produced demographic shifts in which an increasing number of patients have sensory and physical disabilities that may limit their ability to access these new information tools. This study uses automated and manual methods to measure the compliance of 30 popular consumer health websites with the World Wide Web Consortium (W3C) content accessibility guidelines. Among these 30 sites, 22 failed to satisfy at least one W3C Priority-1 accessibility checkpoint, making it impossible for some groups of disabled users to access information from them. All websites failed to satisfy at least one Priority-2 checkpoint, making it difficult for some groups to access information from the sites. These results suggest that accessibility of many consumer health websites to disabled users is very limited. The health informatics community must become more aware of this problem, particularly because many critical accessibility problems may be easily addressed if they are recognized PMID- 15360989 TI - Finding the answers in primary care: information seeking by rural and nonrural clinicians. AB - BACKGROUND: Information systems for rural practice may assume that rural clinicians have different information seeking, but studies have not directly compared rural and nonrural information needs using common methodology. OBJECTIVE: Compare rural and non-rural: 1) information needs; 2) information seeking; 3) effectiveness of information seeking; and 4) use of information resources. DESIGN: Observation and interviews during one half-day of office practice; telephone follow-up 2-10 days later. PARTICIPANTS & SETTING: Primary care physicians (39), nurse practitioners (42), and physician assistants (22) in ambulatory practices in rural and nonrural Oregon. MEASURES: 1) number of questions asked, 2) number of questions pursued, 3) number of questions answered), and 4) use of knowledge resources. RESULTS: Rural clinicians practiced in smaller groups, but were otherwise similar to nonrural clinicians. During half day interviews, clinicians cared for an average of 8.2 patients (95% CI 7.5 - 8.8) and asked an average of 0.83 questions per patient seen (95% CI 0.73 - 0.92). At follow up, they had pursued an average of 47% of their questions (95% CI 40 - 53%), and reported being successful in finding an answer to 77% of those they pursued (95% CI 70 - 84%). There were no statistically significant differences between rural and nonrural clinicians for any of these variables. CONCLUSIONS: Rural and nonrural clinicians had similar information needs, information seeking, knowledge resource use, and effectiveness at finding answers to their questions. Human consultants, digital resources, and library-based resources were less available, but these differences in availability had little impact on their use. PMID- 15360990 TI - A mediated health search portal for trusted medical content. AB - The Internet has become an important source of medical information for both patients and providers. However, it is increasingly difficult to properly access and evaluate the information obtained on the Internet. Search engines are frequently used to find information but suffer either from lack of specificity, or cumbersome requirements for use. Mediated search services have been suggested as a possible alternative. This paper describes one such mediated medical information search service called IMO Health Search. PMID- 15360991 TI - Adapting the human-computer interface for reading literacy and computer skill to facilitate collection of information directly from patients. AB - Clinical information collected directly from patients is critical to the practice of medicine. Past efforts to collect this information using computers have had limited utility because these efforts required users to be facile with the computerized information collecting system. In this paper we describe the design, development, and function of a computer system that uses recent technology to overcome the limitations of previous computer-based data collection tools by adapting the human-computer interface to the native language, reading literacy, and computer skills of the user. Specifically, our system uses a numerical representation of question content, multimedia, and touch screen technology to adapt the computer interface to the native language, reading literacy, and computer literacy of the user. In addition, the system supports health literacy needs throughout the data collection session and provides contextually relevant disease-specific education to users based on their responses to the questions. The system has been successfully used in an academically affiliated family medicine clinic and in an indigent adult medicine clinic. PMID- 15360992 TI - Evaluating consumer informatics: learning from health campaign research. AB - This paper suggests that some conceptual models used in health communication campaigns as well as the "uses and gratifications" approach might be successfully integrated into the evaluation of consumer informatics. These models and tools are especially pertinent when the desired outcomes of media health interventions are therapeutic changes in public knowledge, motivations, attitudes and patient behavior PMID- 15360993 TI - Strategies for supporting consumer health information seeking. AB - Despite a growing number of available Web-based health information resources, consumers continue to face a variety of barriers as they attempt to access these resources. Developing a system that appropriately responds to user queries poses several challenges. Guided by an earlier study that analyzed a large number of queries submitted to ClinicalTrials.gov, we developed a variety of techniques to assist user information seeking. We tested the efficacy of these techniques by submitting the original user queries to our new search engine to determine if these techniques would result in better system performance. Overall, the number of query failures was reduced, but the largest improvement was found in the system's query suggestion capability. For a subset of query failures, the current system was able to cut the earlier failure rate almost in half, in most cases providing a suggestion rather than directly finding records. The techniques described here provide a new approach for responding to user queries. The techniques are tolerant of certain types of errors and provide feedback to assist users in reformulating their queries. PMID- 15360994 TI - Using electronic patient records to inform strategic decision making in primary care. AB - Although absolute risk of death associated with raised blood pressure increases with age, the benefits of treatment are greater in elderly patients. Despite this, the 'rule of halves' particularly applies to this group. We conducted a randomised controlled trial to evaluate different levels of feedback designed to improve identification, treatment and control of elderly hypertensives. Fifty-two general practices were randomly allocated to either: Control (n=19), Audit only feedback (n=16) or Audit plus Strategic feedback, prioritising patients by absolute risk (n=17). Feedback was based on electronic data, annually extracted from practice computer systems. Data were collected for 265,572 patients, 30,345 aged 65-79. The proportion of known hypertensives in each group with BP recorded increased over the study period and the numbers of untreated and uncontrolled patients reduced. There was a significant difference in mean systolic pressure between the Audit plus Strategic and Audit only groups and significantly greater control in the Audit plus Strategic group. Providing patient-specific practice feedback can impact on identification and management of hypertension in the elderly and produce a significant increase in control. PMID- 15360995 TI - Computer interviewing in a primary care office: the patients are ready. AB - Computer patient interviewing has been used since 1968 and must be acceptable to a majority of patients for wide spread use to occur. Computer interviewing is still not used widely in the United States. Potential barriers have not been identified in the literature. METHODS: 150 of 164 (91.5%) eligible patients at a family medicine ambulatory practice were enrolled in a study to evaluate computer interview of cough and sore throat complaints. Subjects were given the choice to have the interview in the waiting or examination room. Telephone interviews were conducted 2-4 weeks later with 143/150 patients (95.3%). RESULTS: 102/150 (68%) of subjects chose the wait-ing room and 48/150 (32%) chose the examination room for the computer interview. 127/143 (88.8%) were willing to use the computer interview for evaluation of cough or sore throat again in the future. 116/143 (81.1%) were willing to use the computer interview for other medical complaints in the future. CONCLUSIONS: Patients are willing to use computer interview-ing and some interviews may be conducted in the waiting room. PMID- 15360996 TI - A patient-controlled journal for an electronic medical record: issues and challenges. AB - Partners Healthcare System, Boston, MA, has developed a patient Web portal that features a patient-controlled electronic "journal" to allow patients to interact with their physician's electronic medical record. Patients can view and respond to health reminders, critique electronic chart information maintained by their doctor's office, enter additional clinical information, and prepare information summaries before an office visit. Creating shared information resources to support a collaborative care model required analysis of the business, architectural, and workflow requirements of the patient-controlled clinical portal and the physician-controlled electronic medical record system. In this paper we describe the challenges in aligning the two systems and serving the different user groups. Coupling the Patient Gateway system, serving over 8700 patients of 90 physicians as of September, 2003, with the Longitudinal Medical Record system, serving over 4000 physicians, has required a clear definition of user goals and workflow, well-defined interfaces, and careful consideration of system assumptions to succeed. PMID- 15360997 TI - Putting E-government to work in healthcare environment: a multiregional project funded by the Italian Innovation & Technology Ministry. AB - In 2002, the Italian Ministry of Innovation promoted a national bid for e government projects. Specifically it allocated a budget of 120 M euro. One of the four project approved in healthcare sector was the "Information, Care ("Assistenza" in Italian) and healthcare Education by the Web" (IAEW), with a global budget of 2580 k euro, partially financed by Ministry with a quota of 830 k euro. The project involves 12 medical structures (both national excellences centers and local regional hospitals) located in two different Region of North Italy, dealing with two different healthcare regional systems (Lombardia and Emilia-Romagna), with potentially 3 millions of users. PMID- 15360998 TI - Towards responsive IT-infrastructures--assessment of a health information system. AB - Marburg University Medical Center has been introducing a comprehensive health information system since 1999, using a single-vendor application framework with an integrated generator tool for the development of clinical applications. To find out if this architecture and our participative software engineering approach can be considered a step towards a responsive infrastructure, we compared the situation after the first deployment phase (basically a holistic approach) with the situation after the system was further developed and adapted to the users' needs using the generator tool approach. We collected system statistics and conducted user satisfaction surveys in 2001/02 and 2003 using standardized measurements. The survey results showed that user involvement as well as system content were judged significantly higher after the second deployment phase. Insofar, we could demonstrate the feasibility of our approach. However, definite statements concerning the superiority of the generator tool approach to other concepts are not yet possible. We will continue our assessment, and we strongly suggest further studies in other institutions introducing comparable clinical functionality. PMID- 15360999 TI - The Hong Kong Hospital Authority's information architecture. AB - Since 1994, the Hospital Authority has been developing and deploying clinical applications at its constituent 39 hospitals and clinics. The Clinical Management System (CMS) is now used by over 4000 doctors and 20000 other clinicians to document and review care. Since 1999, the territory-wide integrated Electronic Patient Record (ePR) has given clinicians a longitudinal view of the data collected through the CMS and its adjunct systems. The ePR currently has nearly 3TB of data covering 44 million episodes for 6.4 million patients. This paper describes the Hospital Authority's Information Architecture, which allows the ePR to accept and integrate any clinical information from any internal or external system. The ePR operates in a high volume and high performance environment, yet only requires low maintenance, while still retaining the information structure and semantics required for advanced applications. PMID- 15361000 TI - National and regional health information infrastructures: making use of information technology to promote access to evidence. AB - The vision for national and regional health information infrastructures (HII) includes provision of a framework that is supportive of access and integration of health information with the goal of improving the health and safety of individuals, public health systems, and nations. Internationally, prominent examples of national and regional HIIs exist that provide a means for achievement of this goal. However, to fully realize benefits, an explicit mechanism is needed for linking national and regional HIIs with existing knowledge, automated processes and evaluation of the ability of HIIs to meet the information needs of primary recipients. Using the United States' Na-tion Health Information Infrastructure (NHII) as an example, the authors describe expansion of the conceptual framework to explicitly acknowledge the role of access to evidence at the overlap between the three dimensions of the NHII to create an "evidence based" link between interrelated components. The role of national measures in setting e-communication goals and evaluating the evolving infrastructure in meeting informational needs of users is discussed. Additionally, automated knowledge management tools such as practice guidelines are presented as a means by which access to critical information is delivered to users, in a format that is appropriate for their health literacy level and that provides adequate support for informed decision making. PMID- 15361001 TI - The RODS Open Source Project: removing a barrier to syndromic surveillance. AB - The goal of the Real-time Outbreak and Disease Surveillance (RODS) Open Source Project is to accelerate deployment of computer-based syndromic surveillance. To this end, the project has released the RODS software under the GNU General Public License and created an organizational structure to catalyze its development. This paper describes the design of the software, requested extensions, and the structure of the development effort. PMID- 15361002 TI - Visualizing the infrastructure of US healthcare using Internet GIS: a community health informatics approach for reducing health disparities. AB - Policy makers and healthcare providers often lack the resources or information to make effective healthcare decisions that impact their communities. Information must therefore be delivered to in a way that maximizes healthcare decision making. This paper examines the infrastructure of U.S. Healthcare using an Internet-based geographic information system (GIS). Internet GIS assists communities in accessing health information via the Internet, thereby leveling the playing field between urban and less affluent rural communities in the United States. A Community Health Informatics approach of incorporating "place" in identifying and reducing health dis-parities and transforming patients to active consumers of health care may ameliorate the impact of devolution and provide a foundation for effective decision support. The Internet GIS includes data visualization tools that provide the public, health care providers, and policy makers with tools to examine socio-economic and demographic factors that impact the distribution of health services PMID- 15361003 TI - Analysis of Web access logs for surveillance of influenza. AB - The purpose of this study was to determine whether the level of influenza in a population correlates with the number of times that internet users access information about influenza on health-related Web sites. We obtained Web access logs from the Healthlink Web site. Web access logs contain information about the user and the information the user accessed, and are maintained electronically by most Web sites, including Healthlink. We developed weekly counts of the number of accesses of selected influenza-related articles on the Healthlink Web site and measured their correlation with traditional influenza surveillance data from the Centers for Disease Control and Prevention (CDC) using the cross-correlation function (CCF). We defined timeliness as the time lag at which the correlation was a maximum. There was a moderately strong correlation between the frequency of influenza-related article accesses and the CDC's traditional surveillance data, but the results on timeliness were inconclusive. With improvements in methods for performing spatial analysis of the data and the continuing increase in Web searching behavior among Americans, Web article access has the potential to become a useful data source for public health early warning systems. PMID- 15361004 TI - The Brazilian health informatics and information policy: building the consensus. AB - This paper describes the construction of the Brazilian Health Information Policy. The Introduction gives an overview of the health informatics scenario in the country and the motivation for the definition of a national policy for the area. The process adopted and the strategies to reach consensus among the different players of the healthcare arena are discussed. The interface with the national health card project and the standards already established are also depicted. The current document and the strategies so far proposed are presented with their respective time table and goals. At the end, a comparison with other national initiatives is drawn. PMID- 15361005 TI - Collection and integration of clinical data for surveillance. AB - OBJECTIVE: The syndromic surveillance project at Public Health-Seattle & King County incorporates several data sources, including emergency department and primary care visit data collected and normalized through an automated mechanism. We describe significant changes made in this "second generation" of our system to improve data quality while complying with privacy and state public health reporting regulations. METHODS/RESULTS: The system uses de-identified visit and patient numbers to assure data accuracy, while shielding patient identity. Presently, we have 124,000 basic visit records (used to generate stratified denominators), and 29,000 surveillance records, from four emergency departments and a primary care clinic network. The system is capable of producing syndrome clustered data sets for analysis. DISCUSSION: We have incorporated data collection techniques such as automated querying, report parsing, and HL7 electronic data interchange. We are expanding the system to include greater population coverage, and developing an understanding how to implement data collections more rapidly at individual hospital sites, as well as how best to prepare the data for analysis. PMID- 15361006 TI - A model of developing medical terms in indigenous languages: a step towards consumer health informatics in South Africa. AB - This paper presents a methodology that has been used in the development of medical terms in the indigenous languages of South Africa. Development of medical terms in indigenous languages is the primary step towards Consumer Health Informatics in the country. This process has been driven by, among others, an emerging paradigm shift in patient care management, whereby patients and other members of the community are encouraged to participate in the process of making treatment or management choices regarding their illnesses or health conditions. In view of this change in the health environment, language and understanding seem to play a major role. Without clear communication and understanding between health providers and the consumers of health services, little can be achieved. Language comprehension is invaluable to empower patients to engage in the decision-making process regarding their health problems. PMID- 15361007 TI - Integration models in health information systems: experiences from the PlugIT project. AB - Different approaches are available for the integration of existing health information systems (HIS) in integration projects. Within the PlugIT project in Finland, we have found it necessary to design and implement integration in a collaborative, multidisciplinary and open way. In this paper, we use some generic integration models and relate them to the methods, solutions and experiences of the project. We summarize the results from nine integration teams, methods development and supporting surveys and studies, and discuss these experiences to provide some guidelines for the HIS integration projects in general. PMID- 15361008 TI - Validating national informatics policy--the importance of operational consensus to influence positive developments. AB - The National Health Service in England faces reorganisation of services on a very regular basis. In 2002 a 'Long Term Review of Health Trends', commissioned by the Chancellor of the Exchequer indicated the need for a substantially larger investment in informatics to support the delivery of better healthcare overall. Subsequent to the issue of this report, the Spending Review confirmed an investment of pound 2.3 Billion (approximately $3.7 Bn) for the period to 2005 for NHS informatics, subject to performance. This paper describes the actions taken by the national representative society (the British Computer Society Health Informatics Committee) to ensure that the views of those in the field were taken into account in facilitating the best possible outcomes from this investment. In addition, the initiative established has confirmed the ongoing priorities for involvement in health informatics, regardless of professional role, in support of healthcare. The outputs and insights gained from two years of this initiative provide useful points for thought about health informatics and health management in other countries and under different models of care to that of the NHS in England. PMID- 15361009 TI - Whole population secondary prevention of coronary heart disease in Scotland: the HEARTS database. AB - Implementing the evidence base for the management of chronic disease is as challenging as discovering which interventions are effective. The HEARTS collaboration (Heart disease Evidence-based Audit and Research in Tayside Scotland) is achieving that goal for the secondary prevention of coronary heart disease (CHD by linking national databases to manually validated hospital and family practice electronic patient records. Specific data from the system is fed back to practices in a facilitated educational process and through the NHS intranet. This paper describes some of the key features of this strategic decision support system. All residents of the Tayside region of Scotland (n=484,013 mid year estimate 2002) are covered by the system. 9,828 patients who have suffered a myocardial infarction(MI) or who have undergone angioplasty or bypass surgery are registered on the system. Improvements in clinical status and prescribing of effective therapies are 5-10% greater than elsewhere in Scotland. PMID- 15361010 TI - Conceptual framework of health indicators: the IDA model. AB - This paper presents a flexible conceptual framework for pub-lic health indicator databases. The model is a multidimensional-hierarchical representation of statistical data describing health and health influencing factors. The main characteristics of the IDA model are the strong discrimination of concepts (categories of entities enumerated in statistical systems) and dimensions (aspects that divide categories). Top level concepts of known data sources (WHO HFA database, OECD Health Data and ECHI) have been compared and a generalized structure had been created which can represent easily and consistently all the top level concepts of the known data sources. The model has been implemented in a prototype system, which demonstrates the feasibility of the approach. PMID- 15361011 TI - EVEREST: an efficient method for verification of digital signatures in real-time teleradiology. AB - The introduction of digital medical images requires a legally binding digital signature that guarantees authenticity and integrity of the image. In real-time teleradiology services, the system is expected to respond very quickly however to verify the signature a considerable amount of time is spent to compute the hash value of the image since the image size might be huge (tens of megabytes). Motivating by this fact, in this paper we propose EVEREST, an efficient methodology for verification. The key observation we have made is that in the traditional verification the processor of the verifying machine is idle (I/O blocked) while the image is downloaded. In EVEREST, to improve the real-time efficiency the receiver can perform most of the hash computation while he is receiving the image itself. One other important advantage of our scheme is the communication efficiency since getting the entire image file is no longer necessary to detect the tampering. PMID- 15361012 TI - We did the right thing: an intervention analysis approach to modeling intervened SARS propagation in Singapore. AB - In this paper, we adopt the Intervention Analysis approach to model an intervened natural process, i.e., propagation of the severe acute respiratory syndrome (SARS) in Singapore, which is affected not only by its own evolutionary history but also by the control measures taken. Using this model, the propagation trend of the epidemic and the effects of different control measures on the outcomes of this epidemic can be simulated and quantitatively analyzed. Based on the model, we have performed an evaluation and sensitivity analysis of the Singapore government's responses to this epidemic. Preliminary results have shown that the control measures taken are effective in controlling the outbreak. PMID- 15361013 TI - Towards interoperability of heterogeneous health databases: application to a tumor samples bank. AB - OBJECTIVES: to define principles and methods that allow heterogeneous database in the health sector to be interoperable. MATERIAL AND METHOD: to design a component based middleware able to provide flexible and efficient means of communication between end-users and databases, and that exploits the standard nomenclatures of the health sector. RESULTS: according to these principles, to implement a prototype of a tumor sample bank in the University hospitals of Marseille, France. DISCUSSION: to discuss the benefits that the approach brings and the progress in prototyping. PMID- 15361014 TI - A Medical Association Hospital centered network for inter-institutional treatment in a regional area, Japan. AB - Munakata Medical Association Hospital is leading the way with its liaison system for electronic medical records and sharing medical information system in Munakata area clinics. A validity experiment was carried out from December 2001 to Autumn 2003 and over 600 patient joined the examination. 90% of the patients surveyed judged it to be convenient about using the system. Medical Image report is most usefull for clinic doctor. The PHS data communication system is easy to install and provides greater flexibility for users over a wider area. Thus, it has proven to be a useful tool for information communication in regional medical care. PMID- 15361015 TI - Predicting survival causes after out of hospital cardiac arrest using data mining method. AB - BACKGROUND: The prognosis of life for patients with heart failure remains poor. By using data mining methods, the purpose of this study was to evaluate the most important criteria for predicting patient survival and to profile patients to estimate their survival chances together with the most appropriate technique for health care. METHODS: Five hundred and thirty three patients who had suffered from cardiac arrest were included in the analysis. We performed classical statistical analysis and data mining analysis using mainly Bayesian networks. RESULTS: The mean age of the 533 patients was 63 (+/- 17) and the sample was composed of 390 (73 %) men and 143 (27 %) women. Cardiac arrest was observed at home for 411 (77 %) patients, in a public place for 62 (12 %) patients and on a public highway for 60 (11 %) patients. The belief network of the variables showed that the probability of remaining alive after heart failure is directly associated to five variables: age, sex, the initial cardiac rhythm, the origin of the heart failure and specialized resuscitation techniques employed. CONCLUSIONS: Data mining methods could help clinicians to predict the survival of patients and then adapt their practices accordingly. This work could be carried out for each medical procedure or medical problem and it would become possible to build a decision tree rapidly with the data of a service or a physician. The comparison between classic analysis and data mining analysis showed us the contribution of the data mining method for sorting variables and quickly conclude on the importance or the impact of the data and variables on the criterion of the study. The main limit of the method is knowledge acquisition and the necessity to gather sufficient data to produce a relevant model. PMID- 15361016 TI - Jogging support system with portable monitoring device and health manage software. AB - At MEDINFO 2001 we reported about an ambulatory biosignal memory device. As an application of this system, we have developed a portable jogging monitoring device and studied its application for health management here. This device is a micro processor based system with some sensors necessary to monitor condition of the user during jogging, that is, a heart rate sensor, a GPS sensor and a physical activity sensor. We also have developed PC based health management software that receives jogging data after jogging and stores it as a long term jogging trend data. From these long term data it feeds back the distance completion of jogging on the virtual course. Moreover it can show the trend of jogging time, distance, average heart rate and the user can know his/her own health condition. PMID- 15361017 TI - A submission model for use in the indexing, searching, and retrieval of distributed pathology case and tissue specimens. AB - This paper describes the Shared Pathology Informatics Network (SPIN) submission model for uploading de-identified XML annotations of pathology case and specimen information to a distributed peer-to-peer network architecture. SPIN use cases, architecture, and technologies, as well as pathology information design is described. With the architecture currently in use by six member institutions, SPIN appears to be a viable, secure methodology to submit pathology information for query and specimen retrieval by investigators PMID- 15361018 TI - Privacy-preserving data releases for health report generation. AB - Regional healthcare initiatives seek to improve the quality of healthcare by collecting, analyzing, and disseminating information about chronic diseases such as diabetes. The data required to support such initiatives comes from several organizations such as insurers, physicians, hospitals, pharmacies and labs each of which gather and maintain data for the purpose of healthcare delivery. In this paper, we focus on mediator-based architectures and the privacy problems that arise in the healthcare context owing to the linkage of information about patients, physicians, and diseases enabled by the mediator. In particular, we examine privacy issues for the two separate steps of the actual data release. First, raw data is released to the (not necessarily trustworthy) mediator and second, the mediator creates and releases the health report. For both steps, we present a technical solution that permits the final report to be useful to the user while respecting the data owners' privacy. PMID- 15361019 TI - Healthcare public key infrastructure (HPKI) and non-profit organization (NPO): essentials for healthcare data exchange. AB - To share healthcare information and to promote cooperation among healthcare providers and customers (patients) under computerized network environment, a non profit organization (NPO), named as OCHIS, was established at Osaka, Japan in 2003. Since security and confidentiality issues on the Internet have been major concerns in the OCHIS, the system has been based on healthcare public key infrastructure (HPKI), and found that there remained problems to be solved technically and operationally. An experimental study was conducted to elucidate the central and the local function in terms of a registration authority and a time stamp authority by contracting with the Ministry of Economics and Trading Industries in 2003. This paper describes the experimental design with NPO and the results of the study concerning message security and HPKI. The developed system has been operated practically in Osaka urban area. PMID- 15361020 TI - Computer simulation of pathogen transmission in the medical intensive care unit: a comparison of two probabilistic methods. AB - The quantitative evaluation of pathogen transmission in the medical intensive care unit (MICU) is difficult given the small number of patients and the complexity and severity of illness. We sought to evaluate the suitability of a probabilistic computer model of our MICU, with which we could rapidly simulate infection control measures and other clinical interventions that would be impossible to perform in the real clinical setting PMID- 15361021 TI - A framework of interruptions in distributed team environments. AB - In this study, we developed a framework for the study of interruptions in distributed team environments from the perspectives of distributed cognition and activity theory. The core of this framework is the Action Coding System (ACS) that provides a language for the description, representation, categorization, and analysis of interruptions at the level of activities that are meaningful for team performance. We demonstrated the practical utility and theoretical significance of ACS in an its application to a real-world, complex, dynamic, and mission critical environment--the Biomedical Engineer (BME) console in the Mission Control Center at NASA's Johnson Space Center. We discussed the potentials of our framework for the design of interruption management systems that could potentially eliminate some interruptions by information redesign, delegate others to autonomous agents, and help human agents to better manage the rest. PMID- 15361022 TI - An inventory of evaluation studies of information technology in health care: trends in evaluation research 1982-2002. AB - During the last years the significance of evaluation studies as well as the interest in adequate methods and approaches for evaluation has grown in medical informatics. In order to put this discussion into the historical context of evaluation research, we conducted a systematic overview on trends in evaluation research of information technology in health care 1982-2002. The inventory is based on a systematic literature search in PubMed. Each of the found 1.035 papers from 1982-2002 was indexed based on a taxonomy coding type of information system, clinical domain, research strategy, evaluation methods, setting and evaluation aspects. We found interesting developments in evaluation research in the last 20 years. For example, their has been a strong shift from medical journals to medical informatics journals. With regard to methods, explanative research and quantitative methods have dominated evaluation studies in the last 20 years. From 1982 to today, the number of lab studies and technical evaluation aspects has declined, while studies focusing on process quality or outcome quality of patient care have increased. Based on our results, we are inclined to talk of a kind of maturation of evaluation studies in medical informatics research. PMID- 15361023 TI - Cognitive evaluation of an innovative psychiatric clinical knowledge enhancement system. AB - Psychiatric Clinical Knowledge Enhancement System (PSYCKES) is an innovative information system that presents patient medication history in tabular and graphical form. The system is designed to support therapeutic decision making. In this paper, we present a multifaceted cognitive evaluation of this system. The evaluation includes a cognitive walkthrough which is a task-analytic method for usability evaluation. We also conducted cognitive studies of two trainee and two attending psychiatrists using the system. One of the attending subjects is presented as a case study. An objective of this research is to characterize the way PSYCKES mediates reasoning. The study found that clinicians were able to use the system effectively to extract and coordinate information and draw appropriate inferences. The expert clinicians were better able to construct a coherent patient representation. The study also documented a few usability problems pertaining to the temporal integration of patient data. PSYCKES is a multifaceted tool that can significantly enhance therapeutic decision making. PMID- 15361024 TI - Differing faculty and housestaff acceptance of an electronic health record one year after implementation. AB - In order to determine whether differences exist between housestaff and faculty physician acceptance of an electronic health record system, we conducted a written survey of attitudes towards new electronic medical record at the University of Illinois at Chicago. We surveyed 330 faculty and housestaff physicians. User acceptance of the EHR was high for both faculty physicians and housestaff. 88.0% of the housestaff and 64.7% of the faculty preferred the EHR over a paper record. Although both housestaff and faculty acceptance of an EHR was high, housestaff showed greater approval ratings than faculty. Central to acceptance of an EHR is conservation of physician time including improving system speed, reducing time spent waiting for a computer to become available, and minimizing time spent documenting care PMID- 15361025 TI - An exploratory study to assess the computer knowledge, attitude and skill among nurses in health care setting of a selected hospital in Ludhiana, Punjab, India. AB - Explorative study conducted to assess and identify deficit areas of computer knowledge, attitudes and skills among nurses working in the hospital and to examine the relationship among these factors. 120 staff nurses were surveyed by systematic random sampling. Computer knowledge, attitudes and skills were measured by a self-structured computer knowledge questionnaire, computer attitude and skill scale respectively. Data analysis showed that the majority 75% staff nurses had good computer knowledge. 100% of nurses had positive attitudes towards computer utilization. 50.8% and 30.8% had average and fair computer skills respectively. No significant correlation was found between nurses' computer knowledge, attitude and skills. The relationships of computer knowledge, attitude and skill were analyzed among nurses with the selected variables like age, sex, designation, years of nursing service, professional qualification, area of nursing service, type of computer training received, frequency of computer usage and monthly family income. Strategies to enhance nurses' computer knowledge, attitudes and skills were proposed. PMID- 15361026 TI - Inter-observer agreement for quality measures applied to online health information. AB - Many quality criteria have been developed to rate the quality of online health information. However, few instruments have been validated for inter-observer reliability. Therefore, we assessed the degree to which two raters agree upon the presence or absence of information based on 22 popularly cited quality criteria on a sample of 21 complementary and alternative medicine websites. Our preliminary analysis showed a poor inter-rater agreement on 10 out of the 22 quality criteria. Therefore, we created operational definitions for each of the criteria, decreased the allowed choices and defined a location to look for the information. As a result 15 out of the 22 quality criteria had a kappa >0.6. We conclude that even with precise definitions some commonly used quality criteria to assess the quality of health information online cannot be reliably assessed. However, inter-rater agreement can be improved by providing precise operational definitions. PMID- 15361027 TI - Differences in the effects of filters on health information retrieval from the Internet in three languages from three countries: a comparative study. AB - We selected twenty search terms on woman's health from various sources and tested them on Google, once with strict filter on and once with filter off. Searches were specified to three countries (Mainland China, Germany, USA), in three languages (Simplified Chinese, German, US English). We found that the proportion of relevant women's health web sites that were blocked was quite high. For the Chinese language web sites originated in China, 72.6% of the blocked web sites were relevant. For the German language web sites originated in Germany, nearly half (49.4%) were relevant. For the US English web sites originated in the US, 95% were relevant. We concluded that people might unknowingly miss potentially important health information due to information filtering. PMID- 15361028 TI - Cancer-related complementary and alternative medicine online: factors affecting information retrieval. AB - Cancer patients increasingly turn to the Internet for health information. As the use of complementary and alternative medicine (CAM) is also increasing, we studied the likelihood that consumers will incidentally encounter CAM information while searching the Internet and the factors that influence CAM information retrieval. We evaluated results retrieved from ten cancer-related searches in six common search engines, and found that 16.2% of 1121 results contained CAM information. Sponsored (i.e. paid) results contained more CAM information than non-sponsored results (38% vs. 7.5%, p<0.001). Sponsored results in the Overture and Google search engines accounted for 51% and 39% of results on the first page. These search engines also retrieved the most CAM web pages. The type of cancer used as the search keyword did not influence the number of CAM-related web pages retrieved. However, the synonyms of cancer used as search keywords differed in their retrieval of CAM web pages (p<0.001). We conclude that clinicians should help consumers recognize sponsored listings and encourage search engines to clearly identify sponsored results. PMID- 15361029 TI - Evaluating the impact of information communication technologies on complex organizational systems: a multi-disciplinary, multi-method framework. AB - The health informatics research community has been undertaking work at the organization-technology intersection for some time now. However there is no one place in the literature which presents a rigorously-defined evaluation framework for use by practitioners and researchers seeking to assess the impact of information and communication technologies on organizational processes and outcomes. There are two main challenges. One is to conceptualize the design features of such an evaluation framework. The second is to specify what data will be gathered and how. This paper aims to address each of these problems. PMID- 15361030 TI - Relationships among different subjective measurements of consumer health information retrieval performance. AB - BACKGROUND: Millions of consumers perform health information retrieval (HIR) online. To better understand the consumers' perspective on HIR performance, we conducted an observation and interview study of 97 health information consumers. METHODS: Consumers were asked to perform HIR tasks and we recorded their view regarding performance using several differ-ent subjective measurements: finding the desired information, usefulness of the information found, satisfaction with the information, and intention to continue searching. Statistical analysis was applied to verify if the multiple subjective measurements were redundant. RESULT: The measurements ranged from slight agreement to no agreement among them. A number of reasons were identified for this lack of agreement. CONCLUSION: Although related, the four subjective measurements of HIR performance are distinct from each other and carried different useful information PMID- 15361031 TI - Patient opinion--EHR assessment from the users perspective. AB - Electronic Health Records (EHR) are becoming an integrated part of modern health care. The introduction of EHR in healthcare has been evaluated by many actors and from many perspectives. However, there is hardly any study exploring the opinion of the ultimate users of the system, the patients. This study aims to provide information about patient values and communication that will be useful in the design of a more patient friendly health system. We have asked patients in three different hospitals about their opinions about EHR, using semi structured individual interviews and focus groups. The patients are open to the use of EHR, but they have some concerns: they want their privacy to be respected, and the systems to be safe from the intrusion from outsiders. These results underline the general societal tendencies of citizen accepting the use of new technology, but being very keen to protect their privacy. PMID- 15361032 TI - Clinical validation of different echocardiographic motion pictures expert group-4 algorythms and compression levels for telemedicine. AB - Tele-echocardiography is not widely used because of lengthy transmission times when using standard Motion Pictures Expert Groups (MPEG)-2 lossy compression algorythms, unless expensive high bandwidth lines are used. We sought to validate the newer MPEG-4 algorythms to allow further reduction in echocardiographic motion video file size. Four cardiologists expert in echocardiography read blindly 165 randomized uncompressed and compressed 2D and color Doppler normal and pathologic motion images. One Digital Video and 3 MPEG-4 compression algorythms were tested, the latter at 3 decreasing compression quality levels (100%, 65% and 40%). Mean diagnostic and image quality scores were computed for each file and compared across the 3 compression levels using uncompressed files as controls. File dimensions decreased from a range of uncompressed 12-83 MB to MPEG-4 0.03-2.3 MB. All algorythms showed mean scores that were not significantly different from uncompressed source, except the MPEG-4 DivX algorythm at the highest selected compression (40%, p=.002). These data support the use of MPEG-4 compression to reduce echocardiographic motion image size for transmission purposes, allowing cost reduction through use of low bandwidth lines. PMID- 15361033 TI - Homecare: the need for cooperative information systems. AB - Assuming responsibility of patients at home (Homecare) is organized around a complex cooperation of partners: health care actors, relatives and helpers, all of whom intervene in the patient's home and who spark off the setting up of cooperative information systems. We present our work relating to such systems within the context of home care. Such activity is based on the collaboration of multiple mobile actors, obtaining information in a multimodal fashion, while taking the job profiles and professional grades of the users into account. Information is obtained from heterogeneous systems. The quality of management of the activity of the various health care actors and of the feedback on information handled at the time of the homecare process will determine how easy it may be to set up homecare as well as the quality of care. In this paper we outline the main stages of our work: grasping the context of homecare and studying co-operative activity from a fundamental point of view but also as applied to homecare. We describe the system proposed for accessing distributed information and for organizing the supervision of the 2 fundamental processes: (1) the LOGISTICAL process (to manage the organization), (2) and the CARE Process (to follow-up the medical or nursing status of the patient), and then we enhance the contribution of mobile technologies in this context. PMID- 15361034 TI - Achieving a "therapeutic dose" of IT. AB - Our health systems face a number of critical challenges over the next several decades, not the least of which is the aging of our baby boomers. Many have suggested that information technologies will be an essential tool, which will increase health system productivity and in so doing will help us manage our way through these challenging times. Unfortunately, little evidence exists supporting the hypothesis that IT creates productivity gains in any sector, let alone health. This paper reviews the "Productivity Paradox" and the challenges associated with measuring the effects that IT may have on health system productivity, and proposes potential directions for new work PMID- 15361035 TI - Numerical model of a nasal septal perforation. AB - This paper describes numerical simulation of airflow in a nose with a nasal septal perforation. Diseased airflow is compared to airflow in a healthy nasal model. The healthy model has been generated from CT scans from the Antrim Area Hospital ENT clinic and is close to being anatomically accurate. The nasal septal perforation has been superimposed on the healthy geometry using image manipulation software. The flow is modeled as laminar, steady state, with the flow rates corresponding to quiet breathing at rest approximately 165 ml/sec. Healthy flow patterns show that the majority of the flow travels close to nasal septum, in the region close to the middle turbinate. In the diseased case, high shear stresses concentrated at the posterior region of the perforation explain bleeding associated with nasal perforations. PMID- 15361036 TI - Institutional decision-making for medical device purchasing: evaluating patient safety. AB - Many medical devices that are currently on the market are sub-optimal for human use, thus contributing to medical errors. This places significant responsibility for device selection on purchasers. This paper describes a retrospective analysis of decision making processes for infusion pump selection in three large hospitals and focuses on patient safety. Through a series of detailed interviews and a study of relevant documentation we characterized the nature of the decision making, patterns of communication, and the roles of different participants. Findings suggest that success of the process is related to effective communication among participants with different expertise and adequate device usability assessment during the process. The paper discusses how information technology could provide support to distributed institutional decision making. PMID- 15361037 TI - A comparison of different heterogeneous proximity functions and Euclidean distance. AB - Proximity functions evaluate distances or similarities between objects. Unlike the Euclidean distance, heterogeneous proximity functions process variables differently according to their scale. The correct evaluation of nominal variables, whose values are unordered, is especially important. We compared five heterogeneous functions with the Euclidean distance to study whether functions sensitive to scale are better than a function assuming the same scale. In addition, we were interested of the relative performance of the five heterogeneous functions. The performance of the functions was measured with a nearest neighbor classifier that was applied to 12 medical data sets characterized with different scales. Unexpectedly, the performance of heterogeneous functions did not differ significantly from that of the Euclidean distance. As expected, significant differences between the Heterogeneous Value Difference Metric (HVDM) and the four value-matching-based heterogeneous functions favored HVDM. Additional research is needed to explain why heterogeneous functions did not outperform the Euclidean distance. PMID- 15361038 TI - Assessing the impact of change in the organization of a technical support system for a health information systems (HIS). AB - OBJECTIVE: Evaluate the impact of the implementation of a new model of Help Desk and technical support in HIS users of the hospital. We carried an anonymous survey on a random sample of 150 users of the new system. The administrative staff (A), doctors (D) and nurses (N) were analysed by strata. We assessed the accessibility both to phone calls and through the institutional intranet; the understanding of the problems; the time taken to fully answer requests; and the degree of satisfaction concerning the change. The results showed that 94%, gained access through the intranet and that it was very satisfactory for the medical group (D80% vs A34% vs N8.7%). The different kinds of users were satisfied with the response time, above all the administrative stratum (A42% vs D38% vs N14%). All of them commented on their satisfaction with the change, above all, the doctors (D68% vs A46% vs N22%), who before this new implementation had expressed dissatisfaction with the old system. CONCLUSIONS: operational changes in the Help Desk contributed to improve how the service was perceived by its users; nevertheless both doctors and nurses required even faster response time. PMID- 15361039 TI - A clinical care costing method. AB - The Clinical Care Costing Method (CCCM for the Home Health Care Classification (HHCC) System provides an innovative way to determine the cost of clinical nursing practice. This costing methodology consists of three major nursing indicators--Care Components, Actions and Outcomes. These three indicators require Clinical Care Pathways (CCP) to document, track, and code clinical care using the HHCC System. The clinical care costs and/or resources are derived from the time and frequencies of the Action Types for the specific nursing interventions performed by the different type of health care providers to achieve the Outcomes and resolve the Care Component that are used to classify nursing diagnoses/patient problems. This method can also be used to deter-mine the reimbursement for nursing care services retrospectively and once validated prospectively. The Clinical Pathway data provide the evidence that the nursing interventions achieve the desired outcomes. PMID- 15361040 TI - Parametric brain MR atlases: standardization for imaging informatics. AB - This paper is focused on the development of normal MR brain atlases of intrinsic MR parameters. These parameters permit quantitative comparisons across imaging studies (as opposed to raw image intensity values) and are important markers of neurological diseases. The development includes fast sequences to generate three parameters (T1: spin-lattice relaxation, T2: spin-spin relaxation, and Diffusion Tensor) covering the whole brain with isotropic and high-resolution images. The analysis of raw data to generate the parametric images is followed by registration algorithms to bring the image studies acquired on normal subjects aligned to a common frame of reference. The registration method includes both linear and non-linear algorithms. Two atlas schemes are discussed: an average atlas and a probabilistic atlas. Initial results on sequence development and registration are presented. The atlases are envisaged as an integral part of an imaging informatics infrastructure that enables image analysis across imaging studies to perform automated image data mining. PMID- 15361041 TI - Potential impact of advanced clinical information technology on healthcare in 2015. AB - Clinical information technologies now sporadically available will soon be in routine clinical use, bringing many changes to healthcare. For example, 1) The next generation Internet; 2) Real-time clinical decision support systems; 3) Off line, population-based systems; 4) Large, integrated, individual patient-level phenotypic and genotypic databases with intelligent data mining capabilities; 5) Wireless, invasive and non-invasive physiologic monitoring devices; 6) Natural Language Processing (NLP) systems; and 7) Mathematical models of complex biological systems have the potential to impact significantly the future healthcare delivery system. While new information management and communication techniques and technologies will reduce many of the inefficiencies and inaccuracies of our present systems, there will be an equal, and potentially far more dangerous, set of unintended consequences. Informatics investigators and health system administrators must focus on the study of what is working and what is not, as well as, on development and testing of the new clinical information management and communication technologies, if we are to be ready for the future. PMID- 15361042 TI - The Informatics Collaboratory: building an online community to support health care informatics students. AB - For the past decade, the School of Nursing has examined the role of computer mediated communication and virtual environments in both health care and educational systems. This recent project, the I-Collaboratory, draws upon previous work and builds on recent studies examining the role of social presence in learning communities. The I-Collaboratory creates a virtual infrastructure to support a collaborative learning environment for Master level students in a web based health care informatics program. This innovative and supportive environment allows students to learn through collaborative inter-actions and have access to a cadre of professionals associated with nursing, informatics and health care. This paper provides an introduction to the concept of a collaboratory and describes the set of principles derived from the literature that serve as a foundation. The process of selecting a web-based platform and the development of the online community based on the results of a need assessment is also detailed. The paper concludes with an outline of the online community components and the plans for evaluation. PMID- 15361043 TI - Inter-document coreference resolution of abnormal findings in radiology documents. AB - In the clinical environment, it is often necessary to track the progression of a condition or various pertinent findings over time. Establishing automatic mechanisms for tracking pertinent findings can aid in the management of a condition as well as provide feedback for treatment outcomes assessment. This work focuses on the challenge of correlating observation of pertinent findings, specifically lung masses, across documents from serial computed tomography examinations for lung cancer patients. A probabilistic model is presented to characterize the likeliness of two observed findings from different documents referring to the same entity. A greedy algorithm is also presented that utilizes the probabilistic model to establish coreference links between findings. Results from a preliminary evaluation of this methodology show a precision of 72% and a recall of 63% for the described inter-document coreference resolution task. PMID- 15361044 TI - An analysis of inpatient nursing communications needs. AB - The health care environment is communications and information intensive. Nurses especially have communications as part of their routine activities, yet little is known about specific nursing communications needs and technologies that might address these needs. This project analyzed the specific communications needs of nurses at Massachusetts General Hospital (MGH) in Boston, MA, through focus group meetings, nursing staff interviews, and direct observation of unit communications. Based on these data, an ideal requirements list for a nursing communication system was created. Data were also gathered and analyzed from units piloting cell phones as nursing communications tools. On non-cell phone units we found that the bulk of communication activity is from the front desk operations associate to the nurse through a sub-optimal overhead paging system that is often unclear or inaudible. The pilot of cellular phones has demonstrated improvements in nursing communications at MGH and there are indications that other emerging technologies will be better able to address the ideal communication needs of nurses. PMID- 15361045 TI - Gathering, structuring and describing information needs in home care: a method for requirements exploration in a "gray area". AB - The aim of our research is to develop a method for "requirements exploration"- gathering, structuring and describing requirements for information systems in previously weakly understood areas. In health care, information systems are usually developed for a single organization, or for information flow from one organization to another. In our case domain, multi-professional and multi organizational home care, this approach is not very helpful. Moreover, home care takes place in "no-man's-land"; outside the organizations' infrastructure, in the customer's home, and technology is used scarcely. In this paper, we describe how information requirements were explored in this scantly understood domain by using an activity-theoretical approach. We also explain how this approach served our purposes. The method proved useful in grasping the network of activities and the information needs. The holistic nature of the method was particularly important, since technology is not the only solution to the needs we discovered. PMID- 15361046 TI - Actions in health care organizations: an ontological analysis. AB - In order to ensure that the information and knowledge needed for the management of healthcare is appropriately shared, human behavior within health care organizations (HCOs) needs to be carefully analyzed. Hence, guidelines, protocols, and messaging standards must be combined with models of resources and processes of patient care that are based on a sound ontology of organizations. This requires a general theory of the ontology of social institutions. Among the many groups attempting to develop efficient ways of sharing information across healthcare systems and organizations is Health Level 7 (HL7). Here I address the question whether HL7 reflects a sound analysis of behavior within HCOs on the basis of a sound ontology of organizations. I then apply ontological principles designed to show how the Reference Information Model (RIM) might be modified in such a way as to support efficient communication of medical information within and between healthcare organizations. PMID- 15361047 TI - A usability study of the PubMed on Tap user interface for PDAs. AB - Healthcare providers are challenged by the availability of information at the point of care. Overcoming these challenges requires access to current, reliable, and accurate information. The purpose of the PubMed on Tap project is to discover and implement design principles to facilitate healthcare practitioners' access to medical information at the point-of-care, related to user interface design, organization and performance. The PubMed on Tap prototype was developed for personal digital assistants (PDAs) as a testbed to study these design principles. Usability testing was used to derive conclusions regarding the functionality of PubMed on Tap. Participant events (n=323) mostly occurred in navigational ability (31%) and perceived functionality (33%). A strong correlation (r=0.87) was observed between total time and the sum of negative events in locating functions and navigational categories. Researchers concluded the basic functionality of the PubMed on Tap interface was valuable to users. However, there was room for improvement in the navigational and functional characteristics, such as placing labels on icons, which affected the users' ability to use the research tool. PMID- 15361048 TI - Design requirements for ubiquitous computing environments for healthcare professionals. AB - Ubiquitous computing environments can support clinical administrative routines in new ways. The aim of such computing approaches is to enhance routine physical work, thus it is important to identify specific design requirements. We studied healthcare professionals in an emergency room and developed the computer augmented environment NOSTOS to support teamwork in that setting. NOSTOS uses digital pens and paper-based media as the primary input interface for data capture and as a means of controlling the system. NOSTOS also includes a digital desk, walk-up displays, and sensor technology that allow the system to track documents and activities in the workplace. We propose a set of requirements and discuss the value of tangible user interfaces for healthcare personnel. Our results suggest that the key requirements are flexibility in terms of system usage and seamless integration between digital and physical components. We also discuss how ubiquitous computing approaches like NOSTOS can be beneficial in the medical workplace. PMID- 15361049 TI - Development and pilot evaluation of user acceptance of advanced mass-gathering emergency medical services PDA support systems. AB - The support systems for the Emergency Medical Services (EMS) in the mass gatherings, such as the local marathon or the large international baseball games, had been underdeveloped. The purposes for this study were to develop triage-based EMS Personal Digital Assistant (PDA) support systems for the mass-gatherings and to evaluate users' perceived ease of use and usefulness of the systems in terms of Davis' Technology Acceptance Model (TAM). The systems were developed based on an established intelligent triage PDA support system and two other forms-the general EMS form from the Taipei EMT and the customer-made Mass Gathering Medical form used by a medical center. 23 nurses and 6 physicians in the medical center, who had ever served in the mass gatherings, were invited to examine the new systems and answered the TAM questionnaire. The results showed that the PDA systems included as many 450 information items inside 42 screens under 6 categories and the great potential of using triage-based PDA systems in the mass gatherings. Overall, most of the subjects agreed with that the systems were easy to use and useful for the mass gatherings, and they were willing to accept the systems. PMID- 15361050 TI - Safe introduction of in-hospital wireless LAN. AB - Insufficient research on electromagnetic interference (EMI) with medical electronic equipment by the signals of wireless LAN has been done. Therefore, electromagnetic compatibility between medical electronic equipment and wireless LAN data communications was done (IEEE802.11a, b, and g). First, to determine if medical electronic equipment is affected by EMI caused by radio waves, we irradiated radio waves to ten types of medical electronic equipment in an electromagnetic anechoic chamber. EMI were observed on three pieces of equipment. Next, to determine if the electromagnetic field emitted by the medical devices might interfere with wireless LAN communication, we measured the electric field intensity. Data analysis showed that the electromagnetic wave emitted by microwave ovens was at almost the same center frequency of the communication channel specified by the IEEE802.11b for wireless LAN. Although the number of medical electronic equipment investigated in this study was small, hospital administrators should consider electromagnetic wave testing of all medical electronic equipment to be used in areas of common wireless LAN use when planning the installation of wireless LAN. PMID- 15361051 TI - PubMed on Tap: discovering design principles for online information delivery to handheld computers. AB - Online access to biomedical information from handheld computers will be a valuable adjunct to other popular medical applications if information delivery systems are designed with handheld computers in mind. The goal of this project is to discover design principles to facilitate practitioners' access to online medical information at the point-of-care. A prototype system was developed to serve as a testbed for this research. Using the testbed, an initial evaluation has yielded several user interface design principles. Continued research is expected to discover additional user interface design principles as well as guidelines for results organization and system performance PMID- 15361052 TI - Ubiquitous computing technology for just-in-time motivation of behavior change. AB - This paper describes a vision of health care where "just-in-time" user interfaces are used to transform people from passive to active consumers of health care. Systems that use computational pattern recognition to detect points of decision, behavior, or consequences automatically can present motivational messages to encourage healthy behavior at just the right time. Further, new ubiquitous computing and mobile computing devices permit information to be conveyed to users at just the right place. In combination, computer systems that present messages at the right time and place can be developed to motivate physical activity and healthy eating. Computational sensing technologies can also be used to measure the impact of the motivational technology on behavior. PMID- 15361053 TI - Remote display performance for wireless healthcare computing. AB - Organizations are beginning to recognize that health care providers are highly mobile and optimal care requires providing access to a large and dynamic body of information wherever the provider and patient are. Remote display protocols (RDP) are one way that organizations are using to deliver healthcare applications to mobile users. While many organizations have begun to use RDPs to deliver real time access to health care information to clinicians, little formal work has been done to evaluate the performance or the effectiveness of thin-client computing with health care applications. This study examines the performance of wireless thin-client tablets with two web-based clinical applications, a text-centric, graphics-poor EMR and a graphic-rich image analysis program. The study compares the performance of two popular RDP implementations, Citrix and Microsoft Remote Desktop, with the performance of a traditional web browser in a wireless environment. For both applications, the RDPs demonstrated both higher speed and reduced bandwidth requirements than the web browser. PMID- 15361054 TI - Participatory design and infrastructures for coordination and communication in interdisciplinary mobile health teams. AB - This paper reports on a current project that explores how wireless and mobile technologies, in this case Personal Digital Assistants (PDAs) and Tablet PCs, may be useful in clinical practices. In particular the support and practices for just in-time coordination and communication among members in highly mobile health teams are focused. Their practice is related to just-in-time access to information for coordination of care and treatment related to, but different from, information traditionally stored in the patients' health records and clinical information systems (CIS). To facilitate discussion and participation among various stakeholders, and between organizational contexts and responsibilities, we developed four video scenarios. The paper describes how these scenarios can be used in an organizational development setting to facilitate analysis and design of tools mindful of the relationships between complex and interwoven infrastructures of communication on one hand and just-in time aspects of a mobile clinical practice on the other. PMID- 15361055 TI - The Clinical Document Architecture (CDA) enables electronic medical records to wireless mobile computing. AB - The Clinical Document Architecture (CDA) has proved to be a valuable and powerful standard for a structured exchange of clinical documents between heterogeneous software systems like a Hospital Information System and a Physician Office System. In this paper we want to show how the CDA can additionally be used in order to enhance the Hospital Information System's functionality: each patient related document contained in or generated from the HIS can be converted to a CDA/XML document. With the XML/XSLT-based transformation methods, those documents can be device-specifically transformed. We use this method to display HIS-content on mobile devices like Personal Digital Assistants (PDAs) by extracting the respective data fields from the HIS database, converting them to a CDA/XML document, which is transformed and sent to the mobile devices using a wireless intranet connection. Preliminary results and users' comments are promising, but further evaluation will be necessary. Our approach shows a generic model how clinical data can be dis-played on different devices independently from the underlying HIS using CDA. PMID- 15361056 TI - Life sign decision support algorithms. AB - There is a pressing need in the military for a system that interprets data from a suite of wearable physiological sensors to infer a soldier's current clinical state on the battlefield. The Warfighter Physiological Status Monitoring (WPSM) concept is envisioned by the US Army to address this need. Life sign detection is a key component. The future WPSM system will consist of a body-worn network of biosensors with a central processing control unit containing firmware for assessing the soldier's physiological status. In the present application, the system will diagnose a Dead, Alive, or Unknown (DAU) physiological state, and this information will be made available to field medics and others over separate communication channels. This paper describes the various modules in the DAU determination algorithms and how they interact with each other, as well as a simulator system built for parametric studies of the overall system. PMID- 15361057 TI - An ethnographic study of collaborative clinical trial protocol writing. AB - Clinical trial protocol documents play an important role in clinical research. However, clinical protocol writing remains a complex and relatively un-studied process. Protocols are often written by teams of people, yet little prior research has captured the problems or analyzed the collaboration support needs of protocol writers. Here we present the results of an initial ethnographic study into the clinical trial protocol writing processes at a representative cooperative clinical trial group funded by National Cancer Institute (NCI). We analyzed the collaborative nature of the writing process, identified common problems, derived information and communication support needs of collaborative clinical protocol writers, and provided recommendations to streamline the process. We believe that this paper contributes useful implications for the design of future collaborative clinical protocol writing tools. PMID- 15361058 TI - Design, implementation and management of a web-based data entry system for ClinicalTrials.gov. AB - We describe the development and deployment of a web-based authoring capability, the first implementation of which is used for data entry and management in support of the ClinicalTrials.gov web site. The system facilitates efficient collection of summary protocol information from multiple geographically-dispersed organizations. We explain the motivation for developing this capability, and cite critical design goals. We then describe system design, implementation and operation, focusing on essential aspects of each. We conclude with a summary of the extent to which we met our stated objectives. PMID- 15361059 TI - Design and development of a search interface for an information gathering tool. AB - In order to design effective and usable search interfaces it is essential to fully understand the characteristics of the users and the tasks they perform. In this paper we describe how to use human-centered approach to design a usable search interface. We first conducted user and task analyses of the application domain--Biomedical Engineers' log notes at NASA Johnson Space Center. From these analyses we identified what functions the users want, the tasks they perform, and a coding system for the vocabulary used by the users to log entries in the log notes. We then implemented a prototype of human-centered search interface by using the results of the user and task analyses and by applying other human centered principles. Finally we discussed the implications of human-centered design for general health information systems. PMID- 15361060 TI - Trial bank publishing: phase I results. AB - BACKGROUND: Randomized clinical trials (RCTs) are an important source of evidence for clinical practice, but finding and applying RCT reports to care is time consuming. Publishing RCTs directly into machine-understandable "trial banks" may allow computers to deliver RCT evidence more selectively and effectively to clinicians. METHODS: Authors of eligible RCTs published in JAMA or the Annals of Internal Medicine between January 2002 and July 2003 were invited to co-publish their trial in RCT Bank, an electronic knowledge base containing details of trial design, execution, and summary results. Trial bank staff used Bank-a-Trial, a web based trial-bank entry tool, to enter information from the manuscript into RCT Bank, obtaining additional information as necessary from the authors. RESULTS: The author participation rate rose from 38% to 76% after the first co-published trial was available as an example. Seven diverse RCTs are now co-published, with 14 in progress. CONCLUSIONS: We have demonstrated proof of concept for co publishing RCTs with leading journals into a structured knowledge base. Phase II of trial bank publishing will introduce direct author submission to RCT Bank. PMID- 15361061 TI - A collaborative clinical trial protocol writing system. AB - Increasing complexity in medicine has caused clinical trial experts with disparate backgrounds from multiple organizations to collaborate when developing clinical trial protocols. Although many protocol-authoring tools provide computerbased decision support to assist in protocol writing, few of them provide sufficient collaboration support for a group of protocol writers. The iterative group writing activities among interdisciplinary clinical trial experts call for advanced tool support. Here we present a web-based protocol writing system with integrated support for collaborative reviewing and collaborative editing. The system uses a shared database to store threaded review comments and version information for electronic protocols. It also captures rich group event information to provide cross-activity awareness and to facilitate self coordination within the collaborative writing team. We believe that our system can help streamline collaborative clinical trial protocol writing processes. PMID- 15361062 TI - Reaction of artemisinin with haemoglobin: implications for antimalarial activity. AB - Elucidation of the principal targets of the action of the antimalarial drug artemisinin is an ongoing pursuit that is important for understanding the action of this drug and for the development of more potent analogues. We have examined the chemical reaction of Hb with artemisinin. The protein-bound haem in Hb has been found to react with artemisinin much faster than is the case with free haem. It appears that the uptake of Hb and the accumulation of artemisinin into the food vacuole, together with the preferred reactivity of artemisinin with haem in Hb, may make Hb the primary target of artemisinin's antimalarial action. Both monoalkylated (HA) and dialkylated (HAA) haem derivatives of artemisinin have been isolated. These 'haemarts' bind to PfHRP II (Plasmodium falciparum histidine rich protein II), inhibiting haemozoin formation, and possess a significantly decreased ability to oxidize ascorbic acid. The accelerated formation of HAA from Hb is expected to decrease the ratio of haem to its alkylated derivatives. The haemarts that are generated from 'haemartoglobins' may bring about the death of malaria parasite by a two-pronged effect of stalling the formation of haemozoin by the competitive inhibition of haem binding to its templates and creating a more reducing environment that is not conducive to the formation of haemozoin. PMID- 15361064 TI - The disintegrin domain of ADAM9: a ligand for multiple beta1 renal integrins. AB - Renal tubular epithelial cells in all nephron segments express a distinct member of the metalloprotease-disintegrin family, ADAM9 (a disintegrin and metalloprotease 9), in a punctate basolateral distribution co-localized to the beta1 integrin chain [Mahimkar, Baricos, Visaya, Pollock and Lovett (2000) J. Am. Soc. Nephrol. 11, 595-603]. Discrete segments of the nephron express several defined beta1 integrins, suggesting that ADAM9 interacts with multiple renal integrins and thereby regulates epithelial cell-matrix interactions. Intact ADAM9 and a series of deletion constructs sequentially lacking the metalloprotease domain and the disintegrin domain were assembled as chimaeras with a C-terminal GFP (green fluorescent protein) tag. Stable expression of the ADAM9/GFP protein on the surface of HEK-293 cells (human embryonic kidney 293 cells) significantly decreased adhesion to types I and IV collagen, vitronectin and laminin, but had little effect on adhesion to fibronectin. Expression of the disintegrin/cysteine rich/GFP construct yielded a similar, but more marked pattern of decreased adhesion. Expression of the cysteine-rich/GFP construct had no effect on adhesion, indicating that the disintegrin domain was responsible for the competitive inhibition of cell-matrix binding. To define the specific renal tubular beta1 integrins interacting with the ADAM9 disintegrin domain, a recombinant GST (glutathione S-transferase)-disintegrin protein was used as a substrate in adhesion assays in the presence or absence of specific integrin blocking antibodies. Inclusion of antibodies to alpha1, alpha3, alpha6, alphav and beta1 blocked adhesion of HEK-293 cells to GST-disintegrin protein. Immobilized GST-disintegrin domain perfused with renal cortical lysates specifically recovered the alpha3, alpha6, alphav and beta1 integrin chains by Western analysis. It is concluded that ADAM9 is a polyvalent ligand, through its disintegrin domain, for multiple renal integrins of the beta1 class. PMID- 15361063 TI - Hepatitis A virus proteinase 3C binding to viral RNA: correlation with substrate binding and enzyme dimerization. AB - Proteinase 3C of hepatitis A virus (HAV) plays a key role in the viral life cycle by generating mature viral proteins from the precursor polyprotein. In addition to its proteolytic activity, 3C binds to viral RNA, and thus influences viral genome replication. In order to investigate the interplay between proteolytic activity and RNA binding at the molecular level, we subjected HAV 3C and three variants carrying mutations of the cysteine residues [C24S (Cys-24-->Ser), C172A and C24S/C172A] to proteolysis assays with peptide substrates, and to surface plasmon resonance binding studies with peptides and viral RNA. We report that the enzyme readily forms dimers via disulphide bridges involving Cys-24. Dissociation constants (K(D)) for peptides were in the millimolar range. The binding kinetics for the peptides were characterized by k(on) and k(off) values of the order of 10(2) M(-1) x s(-1) and 10(-2) to 10(-1) s(-1) respectively. In contrast, 3C binding to immobilized viral RNA, representing the structure of the 5'-terminal domain, followed fast binding kinetics with k(on) and k(off) values beyond the limits of the kinetic resolution of the technique. The affinity of viral RNA depended strongly on the dimerization status of 3C. Whereas monomeric 3C bound to the viral RNA with a K(D) in the millimolar range, dimeric 3C had a significantly increased binding affinity with K(D) values in the micromolar range. A model of the 3C dimer suggests that spatial proximity of the presumed RNA-binding motifs KFRDI is possible. 3C binding to RNA was also promoted in the presence of substrate peptides, indicating co-operativity between RNA binding and protease activity. The data imply that the dual functions of 3C are mutually dependent, and regulate protein and RNA synthesis during the viral life cycle. PMID- 15361065 TI - Functional domain organization of the potato alpha-glucan, water dikinase (GWD): evidence for separate site catalysis as revealed by limited proteolysis and deletion mutants. AB - The potato tuber (Solanum tuberosum) GWD (alpha-glucan, water dikinase) catalyses the phosphorylation of starch by a dikinase-type reaction mechanism in which the beta-phosphate of ATP is transferred to the glucosyl residue of amylopectin. GWD shows sequence similarity to bacterial pyruvate, water dikinase and PPDK (pyruvate, phosphate dikinase). In the present study, we examine the structure function relationship of GWD. Analysis of proteolytic fragments of GWD, in conjunction with peptide microsequencing and the generation of deletion mutants, indicates that GWD is comprised of five discrete domains of 37, 24, 21, 36 and 38 kDa. The catalytic histidine, which mediates the phosphoryl group transfer from ATP to starch, is located on the 36 kDa fragment, whereas the 38 kDa C-terminal fragment contains the ATP-binding site. Binding of the glucan molecule appears to be confined to regions containing the three N-terminal domains. Deletion mutants were generated to investigate the functional interdependency of the putative ATP- and glucan-binding domains. A truncated form of GWD expressing the 36 and 38 kDa C-terminal domains was found to catalyse the E+ATP-->E-P+AMP+P(i) (where P(i) stands for orthophosphate) partial reaction, but not the E-P+glucan-->E+glucan-P partial reaction. CD experiments provided evidence for large structural changes on autophosphorylation of GWD, indicating that GWD employs a swivelling-domain mechanism for enzymic phosphotransfer similar to that seen for PPDK. PMID- 15361066 TI - Human cyclo-oxygenase-1 and an alternative splice variant: contrasts in expression of mRNA, protein and catalytic activities. AB - The two COX (cyclo-oxygenase) isoenzymes COX-1 and -2 catalyse the initial step in the conversion of arachidonic acid into PG (prostaglandin) hormones. The identification of an mRNA transcript encoding a splice variant of human COX-1 was reported more than a decade ago [Diaz, Reginato and Jimenez (1992) J. Biol. Chem. 267, 10816-10822], yet catalytic activity and tissue expression of the corresponding spliced protein remained uncharacterized. The splice variant lacks amino acids 396-432, corresponding to the last 37 amino acids of exon 9 of the gene encoding COX-1. These amino acids form a loop at one side of the peroxidase active site of the protein. We expressed the full-length and spliced COX-1 cDNAs in COS-7 and Sf9 insect cells, and determined the PG-forming activity using incubations with radiolabelled arachidonic acid and HPLC analyses. When expressed in either system, abundant PG formation was observed with the full-length COX-1, whereas the spliced protein did not form any detectable product. Peroxidase activity was readily detected in microsomes prepared from COS-7 cells transfected with COX-1 but not with the splice variant. In reverse transcriptase-PCR experiments, we detected the mRNA for the alternatively spliced and full-length COX-1 in human brain, tonsil and colon tissue, yet we were unable to detect expression of the spliced protein in the same tissues using immunoprecipitation and Western-blot analyses. We conclude that, whereas the mRNA transcript for the spliced COX-1 is present in various human tissues, the corresponding protein is either not formed or subject to rapid proteolytic degradation. PMID- 15361067 TI - Regulation of actin cytoskeleton by mDab1 through N-WASP and ubiquitination of mDab1. AB - Migration of cells is critical to development of the central nervous system. Reelin, which was identified from the reeler mutant mice having a defect in the multilamellar structure of the brain, is thought to be a key signalling molecule that functions as a cue for determination of cell position. mDab1 (mouse Disabled homologue 1) functions downstream of Reelin. However, the mechanism by which mDab1 regulates cell migration during brain development is unknown. In the present paper, we show that mDab1 associates with N-WASP (neuronal Wiskott Aldrich syndrome protein) in vitro and in brains of embryonic mice. mDab1 activates N-WASP directly, and induces actin polymerization through the Arp2/3 (actin-related protein 2/3) complex. mDab1 induces formation of filopodia when it is overexpressed in COS-7 cells. This filopodium formation is dependent on N WASP, because expression of an N-WASP mutant that cannot induce Arp2/3-complex mediated actin polymerization suppressed filopodium formation. The PTB (phosphotyrosine-binding) domain of mDab1 binds to N-WASP via the NRFY (Asn-Arg Phe-Tyr) sequence close to the CRIB (Cdc42/Rac-interactive binding) motif of N WASP and activates N-WASP in vitro. When mDab1 is phosphorylated by Fyn kinase in COS-7 cells, mDab1 is ubiquitinated in a Cbl-dependent manner, and mDab1 does not induce filopodium in the presence of activated Fyn. These findings suggest that mDab1 regulates the actin cytoskeleton through N-WASP, which is negatively regulated by phosphorylation-mediated ubiquitination of mDab1. PMID- 15361068 TI - Characterization of the survival motor neuron (SMN) promoter provides evidence for complex combinatorial regulation in undifferentiated and differentiated P19 cells. AB - There exist two SMN (survival motor neuron) genes in humans, the result of a 500 kb duplication in chromosome 5q13. Deletions/mutations in the SMN1 gene are responsible for childhood spinal muscular atrophy, an autosomal recessive neurodegenerative disorder. While the SMN1 and SMN2 genes are not functionally equivalent, up-regulation of the SMN2 gene represents an important therapeutic target. Consequently, we exploited in silico, in vitro and in vivo approaches to characterize the core human and mouse promoters in undifferentiated and differentiated P19 cells. Phylogenetic comparison revealed four highly conserved regions that contained a number of cis-elements, only some of which were shown to activate/repress SMN promoter activity. Interestingly, the effect of two Sp1 cis elements varied depending on the state of P19 cells and was only observed in combination with a neighbouring Ets cis-element. Electrophoretic mobility-shift assay and in vivo DNA footprinting provided evidence for DNA-protein interactions involving Sp, NF-IL6 and Ets cis-elements, whereas transient transfection experiments revealed complex interactions involving these recognition sites. SMN promoter activity was strongly regulated by an NF-IL6 response element and this regulation was potentiated by a downstream Ets element. In vivo results suggested that the NF-IL6 response must function either via a protein-tethered transactivation mechanism or a transcription factor binding an upstream element. Our results provide strong evidence for complex combinatorial regulation and suggest that the composition or state of the basal transcription complex binding to the SMN promoter is different between undifferentiated and differentiated P19 cells. PMID- 15361069 TI - Activation of serine/threonine protein phosphatase-1 is required for ceramide induced survival of sympathetic neurons. AB - In sympathetic neurons, C6-ceramide, as well as endogenous ceramides, blocks apoptosis elicited by NGF (nerve growth factor) deprivation. The mechanism(s) involved in ceramide-induced neuronal survival are poorly understood. Few direct targets for the diverse cellular effects of ceramide have been identified. Amongst those proposed is PP-1c, the catalytic subunit of serine/threonine PP-1 (protein phosphatase-1). Here, we present the first evidence of PP-1c activation by ceramide in live cells, namely NGF-deprived sympathetic neurons. We first determined PP activity in cellular lysates from sympathetic neurons treated with exogenous ceramide and demonstrated a 2-3-fold increase in PP activity. PP activation was completely blocked by the addition of the specific type-1 PP inhibitor protein I-2 as well as by tautomycin, but unaffected by 2 nM okadaic acid, strongly indicating that the ceramide-activated phosphatase activity was PP 1c. Inhibition of PP activity by phosphatidic acid (which has been reported to be a selective inhibitor of PP-1c) and tautomycin (a PP-1 and PP-2A inhibitor), but not by 10 nM okadaic acid, abolished the anti-apoptotic effect of ceramide in NGF deprived neurons, suggesting that activation of PP-1c is required for ceramide induced neuronal survival. Ceramide was able to prevent pRb (retinoblastoma gene product) hyperphosphorylation by a mechanism dependent on PP-1c activation, suggesting that two consequences of NGF deprivation in sympathetic neurons are inhibition of PP-1c and subsequent hyperphosphorylation of pRb protein. These findings suggest a novel mechanism for ceramide-induced survival, and implicate the involvement of PPs in apoptosis induced by NGF deprivation. PMID- 15361070 TI - Human recombinant membrane-bound aminopeptidase P: production of a soluble form and characterization using novel, internally quenched fluorescent substrates. AB - APP (aminopeptidase P) has the unique ability to cleave the N-terminal amino acid residue from peptides exhibiting a proline at P(1)'. Despite its putative involvement in the processing of bioactive peptides, among them the kinins, little is known about the physiological roles of both human forms of APP. The purpose of the present study is first to engineer and characterize a secreted form of hmAPP (human membrane-bound APP). Our biochemical analysis has shown that the expressed glycosylated protein is fully functional, and exhibits enzymic parameters similar to those described previously for mAPP purified from porcine or bovine lungs or expressed from a porcine clone. This soluble form of hmAPP cross-reacts with a polyclonal antiserum raised against a 469-amino-acid hmAPP fragment produced in Escherichia coli. Secondly, we synthesized three internally quenched fluorescent peptide substrates that exhibit a similar affinity for the enzyme than its natural substrates, the kinins, and a higher affinity compared with the tripeptide Arg-Pro-Pro used until now for the quantification of APP in biological samples. These new substrates represent a helpful analytical tool for rapid and reliable screening of patients susceptible to adverse reactions associated with angiotensin-converting enzyme inhibitors or novel vasopeptidase (mixed angiotensin-converting enzyme/neprilysin) inhibitors. PMID- 15361071 TI - Heterogeneous nuclear ribonucleoprotein K represses transcription from a cytosine/thymidine-rich element in the osteocalcin promoter. AB - HnRNP K (heterogeneous nuclear ribonucleoprotein K) was biochemically purified from a screen of proteins co-purifying with binding activity to the osteocalcin promoter. We identify hnRNP K as a novel repressor of osteocalcin gene transcription. Overexpression of hnRNP K lowers the expression of osteocalcin mRNA by 5-fold. Furthermore, luciferase reporter assays demonstrate that overexpression of hnRNP K represses osteocalcin transcription from a CT (cytosine/thymidine)-rich element in the proximal promoter. Electrophoretic mobility-shift analysis reveals that recombinant hnRNP K binds to the CT-rich element, but binds ss (single-stranded), rather than ds (double-stranded) oligonucleotide probes. Accordingly, hnRNP K antibody can supershift a binding activity present in nuclear extracts using ss sense, but not antisense or ds oligonucleotides corresponding to the CT-rich -95 to -47 osteocalcin promoter. Importantly, addition of recombinant hnRNP K to ROS 17/2.8 nuclear extract disrupts formation of a DNA-protein complex on ds CT element oligonucleotides. This action is mutually exclusive with hnRNP K's ability to bind ss DNA. These results demonstrate that hnRNPK, although co-purified with a dsDNA-binding activity, does not itself bind dsDNA. Rather, hnRNP K represses osteocalcin gene transcription by inhibiting the formation of a transcriptional complex on the CT element of the osteocalcin promoter. PMID- 15361072 TI - Structural diversity and specific distribution of O-glycans in normal human mucins along the intestinal tract. AB - Purified human mucins from different parts of the intestinal tract (ileum, cecum, transverse and sigmoid colon and rectum) were isolated from two individuals with blood group ALe(b) (A-Lewis(b)). After alkaline borohydride treatment the released oligosaccharides were structurally characterized by nano-ESI Q-TOF MS/MS (electrospray ionization quadrupole time-of-flight tandem MS) without prior fractionation or derivatization. More than 100 different oligosaccharides, with up to ten monosaccharide residues, were identified using this technique. Oligosaccharides based on core 3 structures, GlcNAc(beta1-3)GalNAc (where GlcNAc is N-acetyl-D-glucosamine and GalNAc is N-acetylgalactosamine), were widely distributed in human intestinal mucins. Core 5 structures, GalNAc(alpha1 3)GalNAc, were also recovered in all fractions. Moreover, a comparison of the oligosaccharide repertoire, with respect to size, diversity and expression of glycans and terminal epitopes, showed a high level of mucin-specific glycosylation: highly fucosylated glycans, found specifically in the small intestine, were mainly based on core 4 structures, GlcNAc-(beta1-3)[GlcNAc(beta1 6)]GalNAc, whereas the sulpho-Le(X) determinant carrying core 2 glycans, Gal(beta1-3)[GlcNAc(beta1-6)]-GalNAc (where Gal is galactose), was recovered mainly in the distal colon. Blood group H and A antigenic determinants were present exclusively in the ileum and cecum, whereas blood group Sd(a)/Cad related epitopes, GalNAc(beta1-4)[NeuAc(alpha2-3)]Gal (where NeuAc is N acetylneuraminate), were found to increase along the length of the colon. Our findings suggest that mucins create an enormous repertoire of potential binding sites for micro-organisms that could explain the regio-specific colonization of bacteria in the human intestinal tract. PMID- 15361073 TI - ADP-ribosylation of integrin alpha7 modulates the binding of integrin alpha7beta1 to laminin. AB - The extracellular domain of integrin alpha7 is ADP-ribosylated by an arginine specific ecto-ADP-ribosyltransferase after adding exogenous NAD+ to intact C2C12 skeletal muscle cells. The effect of ADP-ribosylation on the structure or function of integrin alpha7beta1 has not been explored. In the present study, we show that ADP-ribosylation of integrin alpha7 takes place exclusively in differentiated myotubes and that this post-translational modification modulates the affinity of alpha7beta1 dimer for its ligand, laminin. ADP-ribosylation in the 37-kDa 'stalk' region of alpha7 that takes place at micromolar NAD+ concentrations increases the binding of the alpha7beta1 dimer to laminin. Increased in vitro binding of integrin alpha7beta1 to laminin after ADP ribosylation of the 37-kDa fragment of alpha7 requires the presence of Mn2+ and it is not observed in the presence of Mg2+. In contrast, ADP-ribosylation of the 63-kDa N-terminal region comprising the ligand-binding site of alpha7 that occurs at approx. 100 microM NAD+ inhibits the binding of integrin alpha7beta1 to laminin. Furthermore, incubation of C2C12 myotubes with NAD+ increases the expression of an epitope on integrin beta1 subunit recognized by monoclonal antibody 9EG7. We discuss our results based on the current models of integrin activation. We also hypothesize that ADP-ribosylation may represent a mechanism of regulation of integrin alpha7beta1 function in myofibres in vivo when the continuity of the membrane is compromised and NAD+ is available as a substrate for ecto-ADP-ribosylation. PMID- 15361074 TI - Ribosomal protein L7a binds RNA through two distinct RNA-binding domains. AB - The human ribosomal protein L7a is a component of the major ribosomal subunit. We previously identified three nuclear-localization-competent domains within L7a, and demonstrated that the domain defined by aa (amino acids) 52-100 is necessary, although not sufficient, to target the L7a protein to the nucleoli. We now demonstrate that L7a interacts in vitro with a presumably G-rich RNA structure, which has yet to be defined. We also demonstrate that the L7a protein contains two RNA-binding domains: one encompassing aa 52-100 (RNAB1) and the other encompassing aa 101-161 (RNAB2). RNAB1 does not contain any known nucleic-acid binding motif, and may thus represent a new class of such motifs. On the other hand, a specific region of RNAB2 is highly conserved in several other protein components of the ribonucleoprotein complex. We have investigated the topology of the L7a-RNA complex using a recombinant form of the protein domain that encompasses residues 101-161 and a 30mer poly(G) oligonucleotide. Limited proteolysis and cross-linking experiments, and mass spectral analyses of the recombinant protein domain and its complex with poly(G) revealed the RNA-binding region. PMID- 15361076 TI - Helicobacter pylori HP1034 (ylxH) is required for motility. AB - BACKGROUND: Helicobacter pylori motility is essential for the colonization and persistence in the human gastric mucosa. So far, more than 50 genes have been described to play a role in flagellar biosynthesis. H. pylori YlxH (HP1034) is annotated as an ATP-binding protein. However, H. pylori YlxH shows similarity to proteins involved in the flagellar biosynthesis of other bacterial species. Moreover, H. pylori ylxH is found adjacent to genes involved in flagellar biosynthesis in the sequenced genomes of H. pylori 26695 and J99. We therefore aimed to determine the role of YlxH in H. pylori motility. MATERIALS AND METHODS: Motility, flagellar biosynthesis and transcriptional regulation of genes encoding flagellar proteins was compared between H. pylori 11A and a knockout of ylxH in H. pylori 11A. RESULTS: The ylxH knockout in H. pylori 11A was nonmotile on soft agar plates, whereas H. pylori 11A was motile. Furthermore, the H. pylori 11A ylxH knockout lacked flagella, while H. pylori 11A possessed two to three flagella. Transcription of H. pylori flaG (HP0751), fliM (HP1031) and fliA (HP1032) was reduced in the H. pylori 11A ylxH(;) knockout, whereas transcription of flaA (HP0601) was not altered. However, Western blot analysis showed substantially reduced amounts of the major flagellin subunit FlaA in the H. pylori 11A ylxH knockout compared to H. pylori 11A. CONCLUSIONS: H. pylori YlxH is essential for the assembly of flagella and hence for the motility of H. pylori. PMID- 15361075 TI - Effect of H. pylori on the expression of TRAIL, FasL and their receptor subtypes in human gastric epithelial cells and their role in apoptosis. AB - BACKGROUND AND AIMS: In the human stomach expression of TNF-related apoptosis inducing ligand (TRAIL) and its receptors and the modulatory role of Helicobacter pylori are not well described. Therefore, we investigated the effect of H. pylori on the expression of TRAIL, FasL and their receptors (TRAIL-R1-R4, Fas) in gastric epithelial cells and examined their role in apoptosis. MATERIALS AND METHODS: mRNA and protein expression of TRAIL, FasL and their receptors were analyzed in human gastric epithelial cells using RT-PCR, Western blot, and immunohistochemistry. Gastric epithelial cells were incubated with FasL, TRAIL and/or H. pylori, and effects on expression, cell viability and epithelial apoptosis were monitored. Apoptosis was analyzed by histone ELISA, DAPI staining and immunohistochemistry. RESULTS: TRAIL, FasL and their receptor subtypes were expressed in human gastric mucosa, gastric epithelial cell primary cultures and gastric cancer cells. TRAIL, FasL and H. pylori caused a time- and concentration dependent induction of DNA fragmentation in gastric cancer cells with synergistic effects. In addition, H. pylori caused a selective up-regulation of TRAIL, TRAIL R1 and Fas mRNA and protein expression in gastric cancer cells. CONCLUSIONS: Next to FasL and Fas, TRAIL and all of its receptor subtypes are expressed in the human stomach and differentially modulated by H. pylori. TRAIL, FasL and H. pylori show complex interaction mediating apoptosis in human gastric epithelial cells. These findings might be important for the understanding of gastric epithelial cell kinetics in patients with H. pylori infection. PMID- 15361077 TI - Identification of a novel mutation affecting domain V of the 23S rRNA gene in Helicobacter pylori. AB - BACKGROUND: This study analyzes clarithromycin resistance status and 23S rRNA gene mutations in Helicobacter pylori strains from Central Italian patients. MATERIALS AND METHODS: H. pylori strains from 235 dyspeptic patients (205 with no history of clarithromycin exposure and 30 referred for failure of eradication therapy) were tested for clarithromycin resistance by screening agar method and E test. Resistant strains were analyzed for mutations of the 23S rRNA gene by PCR RFLP and sequencing. RESULTS: Primary resistance was observed in strains from 43/205 (21%) patients with no history of clarithromycin exposure and secondary resistance in 30/30 (100%) strains from previously treated patients. A single mutant strain was detected in 54/73 (74%) cases, a mixture of one or more mutant(s) plus the wild type in the remaining 19/73 (26%) cases. One 23S rRNA gene mutation (A-->T transversion at nucleotide 2144) in the peptidyltransferase region of domain V was novel. CONCLUSIONS: This study shows: (a) a high prevalence of H. pylori strains with primary or secondary clarithromycin resistance in an urban area of Central Italy; (b) colonization by both mutant and wild-type H. pylori in the same patient; (c) a novel variant of the H. pylori 23S rRNA gene. PMID- 15361078 TI - The rdxA gene plays a more major role than frxA gene mutation in high-level metronidazole resistance of Helicobacter pylori in Taiwan. AB - BACKGROUND: Metronidazole-resistant H. pylori associating with mutations of rdxA or frxA is still a debated topic. This study investigates whether rdxA and frxA mutations of H. pylori accounted for the high MIC value (>/= 64 micro g/ml) of metronidazole (Mtz). MATERIAL AND METHODS: From 126 clinical H. pylori isolates, we examined 14 Mtz-sensitive, 18 Mtz-resistant H. pylori, and eight pairs of Mtz sensitive and Mtz-resistant colonies simultaneously present within a single gastric biopsy. The paired strains from one single biopsy were proven identical by PCR-RFLP. MICs of Mtz were checked by the E-test and agar dilution method. The mutations of rdxA and frxA sequencing were matched with the Mtz-susceptible ATCC 26695 and J99. RESULTS: There were 89% (16/18) of Mtz-resistant isolates with mutation of RdxA. Half of the 14 Mtz-sensitive strains, all without mutation of RdxA, still contained truncation of FrxA. Within the paired isolates from a single biopsy, rdxA mutation (86%) was more common than frxA mutation (43%) in those isolates with high-level Mtz-resistant H. pylori. RdxA truncation was more prevalent in Mtz-resistant strains with high MICs than in those with low to moderate MICs (75% vs. 20%, p =.01, OR: 12, 95% CI: 1.8-81.7). CONCLUSION: Mutations in the rdxA gene rather than the frxA gene generally determine a high MIC level of Mtz-resistant H. pylori in Taiwan. PMID- 15361079 TI - Experimental Helicobacter felis infection in transgenic mice expressing human group IIA phospholipase A2. AB - BACKGROUND: Both various virulence factors of Helicobacter pylori and host factors influence the clinical outcome of H. pylori infection. In animal experiments with Helicobacter felis, large variations in the severity of disease have been observed between different mouse strains infected with a single isolate of H. felis. C57BL/6 J mouse strain that lacks the expression of group IIA phospholipase A2 has been shown to develop more severe gastric inflammation than other mouse strains. Thus, group IIA phospholipase A2 has been suggested to play a role in regulating inflammation in gastric mucosa. The aim of this study was to examine the possible role of group IIA phospholipase A2 in experimental Helicobacter infection. MATERIALS AND METHODS: Transgenic mice expressing human group IIA phospholipase A2 and their group IIA phospholipase A2 deficient nontransgenic C57BL/6 J littermates were infected with H. felis. The mice were killed 3, 8, and 19 weeks after inoculation of bacteria to determine the histopathological changes in gastric mucosa. RESULTS: The infected mice developed chronic inflammation in gastric mucosa. We found no differences in the colonization of bacteria between transgenic and nontransgenic mice. At 3 and 8 weeks, no difference was found in the severity of inflammation between the two groups. Nineteen weeks after the administration of bacteria the inflammation was more marked in nontransgenic than transgenic mice. Group IIA phospholipase A2 was expressed by in situ hybridization in the neck cells of the glandular stomach in transgenic mice. CONCLUSIONS: The results of the present study suggest that the endogenous expression of group IIA phospholipase A2 diminishes chronic inflammation in gastric mucosa in experimental H. felis infection in mice. PMID- 15361080 TI - Open, randomized multicenter comparative trial of rabeprazole, ofloxacin and amoxicillin therapy for Helicobacter pylori eradication: 7 vs. 14 day treatment. AB - BACKGROUND: Because of the increasing resistance to clarithromycin and metronidazole, two of the antibiotics used for the eradication of Helicobacter pylori, new therapeutic alternatives are needed. The aim of this study was to determine the efficacy of a randomized, comparative trial of 7 vs. 14-day triple treatment with rabeprazole, ofloxacin and amoxicillin for H. pylori eradication. MATERIAL AND METHODS: The present authors studied 76 dyspeptic patients infected with H. pylori diagnosed by both histology and a rapid urease test. Patients were randomized to receive rabeprazole (20 mg b.i.d.), plus ofloxacin (400 mg b.i.d.) and amoxicillin (1000 mg b.i.d.) for 7 days (group 1) vs. 14 days (group 2) and were followed by 6 weeks. Eradication was assessed 4 weeks after completing the course of study treatment by the (14)C-urea breath test. Per protocol and intention-to-treat eradication rates were determined. RESULTS: For the intention to treat analysis, the eradication rate was 62.2% for group 1 and 92.3% for group 2 (p =.004). For the per protocol analysis, eradication rate for group 1 was 63.9% and for group 2 was 97.3% (p =.001). CONCLUSIONS: Triple therapy with rabeprazole, amoxicillin and ofloxacin by 14 days was efficient for H. pylori eradication and therefore deserves further study. The same regimen prescribed for 7 days had a significantly lower and unacceptable cure rate and should not be used. PMID- 15361081 TI - Selective and effective bactericidal activity of the cobalt (II) cation against Helicobacter pylori. AB - BACKGROUND: Although the anti-Helicobacter pylori activity of bismuth is well established, the therapeutic potential of other metal ions against the organism is not known. MATERIALS AND METHODS: We measured the minimum inhibitory concentrations of a series of metal ions, including several cobalt (II) compounds against four type strains and seven clinical isolates of H. pylori using three standard broth culture media and a defined medium. Other intestinal bacteria were also investigated for specificity of action. RESULTS: Cobalt chloride had marked activity against H. pylori (minimum inhibitory concentration range was 0.03-1.0 mg/l). The effect was specific because other transition metals had no effect and other intestinal bacteria were not affected by cobalt chloride. Activity was attributable to free cobalt ions as ligands inhibited activity in proportion to their affinity for the ions. Inhibition of cobalt activity was also observed in the presence of nickel, in a dose dependent fashion. However, cobalt activity was not directed towards the nickel-dependent urease enzyme because its effect was similar in wild-type and urease negative mutant strains of H. pylori. Finally, the viability of H. pylori was reduced at the same rate with 2 mg/l cobalt as with 1 mg/l amoxicillin. CONCLUSIONS: Cobalt competes for nickel in its acquisition by H. pylori, but mediates toxicity in a nonurease dependent fashion. As cobalt MIC is similar to some antibiotics and 10 to a hundred times lower than for bismuth, cobalt may represent an effective form of therapy for H. pylori infection. PMID- 15361082 TI - Melanoidin, a food protein-derived advanced maillard reaction product, suppresses Helicobacter pylori in vitro and in vivo. AB - BACKGROUND: Extracellular urease proteins located on the surface of Helicobacter pylori are gastric mucin-targeted adhesins, which play an important role in infection and colonization to the host. In this study we have determined the inhibitory activity of a variety of melanoidins, protein-derived advanced Maillard reaction products, ubiquitously found in heat-treated foods, on urease gastric mucin adhesion. In addition, we have determined the anticolonization effect of melanoidin I, prepared by the Maillard reaction between casein and lactose, in an animal model and in human subjects infected with this bacterium. METHODS: The inhibitory activity of each compound was determined by a competitive binding assay of labeled gastric mucin to plate-immobilized urease. Melanoidin I was used in an in vivo trial using euthymic hairless mice as an infection model. Melanoidin I was consumed for 8 weeks by subjects infected with H. pylori. The [(13)C] urease breath test and H. pylori-specific antigen in the stool (HpSA) test were performed on subjects at week 0 and week 8. RESULTS: A variety of food protein-derived melanoidins strongly inhibited urease-gastric mucin adhesion in the concentration range of 10 micro g/ml to 100 micro g/ml. In particular, melanoidin I significantly (p <.05) suppressed colonization of H. pylori in mice when given for 10 weeks via the diets. Eight weeks daily intake of 3 g melanoidin I significantly (p <.05) decreased the optical density of HpSA in subjects. CONCLUSION: Foods containing protein-derived melanoidins may be an alternative to antibiotic-based therapy to prevent H. pylori that combines safety, ease of administration and efficacy. PMID- 15361083 TI - Role of noninvasive tests (C-urea breath test and stool antigen test) as additional tools in diagnosis of Helicobacter pylori infection in patients with atrophic body gastritis. AB - BACKGROUND: Detection of Helicobacter pylori infection in atrophic body gastritis (ABG) is difficult, as during progression of body atrophy, H. pylori disappears. AIM: To increase the diagnostic yield of detection of active H. pylori infection in atrophic body gastritis patients by using noninvasive tests such as (13)C-Urea Breath Test ((13)C-UBT) and H. pylori stool antigen test (HpSA) would be useful. PATIENTS: 27 consecutive patients with newly-diagnosed atrophic body gastritis (19F/7M, age 27-73 years). METHODS: Gastroscopy with biopsies (antrum n = 3, body n = 3) and histology according to updated Sydney system, H. pylori IgG serology, (13)C-UBT, and HpSA. RESULTS: All tests used in the diagnosis of H. pylori infection were in agreement in 9/27 atrophic body gastritis patients (33.3%), being all positive in four (14.8%) and all negative in five patients (18.5%). Ten of the 27 (37%) patients were Giemsa stain-positive and serology-positive (group I). Seventeen of the 27 (63%) patients were Giemsa stain-negative: 5/17 with positive serology (group II) and 12/17 with negative serology (group III). In group I, 5/10 (50%) were (13)C-UBT positive and 4/10 (40%) HpSA positive. In group II, two patients were (13)C-UBT positive, but all were HpSA negative. Also in group III, all patients were HpSA negative, but one had a positive (13)C-UBT. CONCLUSIONS: In atrophic body gastritis patients, neither (13)C-UBT nor HpSA per se add useful information regarding active H. pylori infection, but these noninvasive tests may be important in combination with histology and serology to define the H. pylori status in some atrophic body gastritis patients. PMID- 15361084 TI - Characteristics of Helicobacter pylori-induced gastritis and the effect of H. pylori eradication in patients with chronic idiopathic thrombocytopenic purpura. AB - BACKGROUND: The association between Helicobacter pylori infection and idiopathic thrombocytopenic purpura (ITP) has been reported widely. We investigated the prevalence of H. pylori infection, its virulence profile and the effectiveness of its eradication in patients with ITP. MATERIALS AND METHODS: Twenty patients with ITP, 20 with peptic ulcer (10 gastric ulcer (GU), 10 duodenal ulcer (DU)) and 20 with NUD were studied. The virulence profile of the strains was assessed by genotyping for cagA, vacA, iceA, and hpyIIIR/hrgA and by assaying for IL-8 and DNA fragmentation after incubation with AGS cells. Infected patients and two uninfected ITP patients received triple therapy and platelets were counted before and 1 month, 6 months, 1 year, and 2 years after eradication therapy. RESULTS: H. pylori infection was found in 17 ITP (85%), 20 ulcer (100%) and 13 NUD (65%) patients. Biopsies and strains were collected from five ITP, 20 ulcer and 13 NUD patients. The ITP patients had a pangastritis or corpus-predominant gastritis pattern. All H. pylori isolates, from ITP, ulcer and NUD patients, were cagA(+) and vacA s1/m1, and did not differ in levels of IL-8 induction or DNA fragmentation. Fifteen ITP (88%) and 17 ulcer (85%) patients had successful eradication of H. pylori. Ten of these 15 (67%) H. pylori-eradicated ITP patients had platelet recovery. There was no significant change in platelet count in the two ITP patients in whom eradication failed or in the two originally H. pylori uninfected ITP patients, or in the treated ulcer patients. Age at onset of ITP was the main determinant of platelet recovery: 100% of patients diagnosed after the age of 60 recovered compared with only 22% of those diagnosed before 50. CONCLUSIONS: H. pylori-infected ITP patients have a corpus-predominant pattern of gastritis but the virulence profile of their strains does not differ from that of ulcer or NUD patients. Eradication of H. pylori infection is a good therapeutic option for some patients with chronic ITP, especially for those who develop ITP in older age. PMID- 15361085 TI - Molecular epidemiology and outcome of Helicobacter pylori infection in Thailand: a cultural cross roads. AB - ABSTRACT Background. Thailand is at the cultural cross roads between East and South Asia. It has been suggested that this is also the region where the predominant Helicobacter pylori (H. pylori) genotype changes from East Asian to South Asian. Methods. We compared the molecular epidemiology and outcome of H. pylori infections among different ethnic groups in Thailand (Thai, Thai-Chinese and Chinese). H. pylori isolates were genotyped by polymerase chain reaction based on cagA, cag right end junction and vacA genotypes. Results. Ninety-eight isolates from 38 ethnic Thai, 20 ethnic Chinese and 40 Thai-Chinese were categorized into East Asian (45%), South/Central Asian (26%), Western (1%) or mixed type (29%). The East Asian genotype was the most common among Chinese (85%) and Thai-Chinese (55%) (p <.01 compared to ethnic Thai). The ethnicity of the mother among mixed Thai-Chinese marriages predicted the genotype of the child's H. pylori (e.g. when the mother was Chinese, 84% had East Asian type vs. 29% when the mother was Thai) (p <.001). Gastric cancer was common among ethnic Chinese with East Asian genotype (e.g. all Chinese with gastric cancer or peptic ulcer disease had East Asian genotype, whereas only 40% of Chinese with gastritis had this genotype). Conclusions. Immigration, intermarriage and the variety of H. pylori genotypes in Thailand suggest that Thailand is an ideal site for epidemiological studies attempting to relate H. pylori genotypes and host factors to outcome. Our data also support the hypothesis that the primary caretaker of the children is most likely the source of the infection. PMID- 15361086 TI - Detection of Helicobacter ganmani-like 16S rDNA in pediatric liver tissue. AB - BACKGROUND: To determine the presence of Helicobacter species in the liver biopsy specimens from children with various chronic liver diseases as data in adult literature suggests a possible role of these bacteria in their pathogenesis. MATERIALS AND METHODS: Paraffin sections of 61 liver biopsies of pediatric patients with miscellaneous diseases and autopsy liver tissue from 10 control subjects with no evidence of preexisting liver disease were examined for the presence of Helicobacter species by a genus-specific seminested polymerase chain reaction (PCR) assay. PCR-products of positive samples were further characterized by denaturing gradient gel electrophoresis (DGGE) and DNA-sequence analysis. Based on those results, a seminested PCR assay for H. ganmani was developed and applied to the samples. RESULTS: On analysis, 40/61 patient samples were positive in the genus-specific Helicobacter PCR and 4/10 from the control group. The nucleotide sequences of 16S rDNA fragments were 99-100% similar to mainly Helicobacter sp. 'liver' and H. ganmani. PCR-products similar to H. canis and H. bilis were also found. The 16S rDNAs of control specimens showed similarity to Helicobacter sp. 'liver'. In the H. ganmani-specific PCR analysis 19 patients, but none of the controls, were positive. CONCLUSIONS: Amplified Helicobacter 16S rDNAs were related to Helicobacter sp. 'liver' or H. ganmani in liver biopsy specimens of pediatric patients. The possible significance of Helicobacter species in pediatric liver diseases needs to be evaluated further in prospective studies. PMID- 15361088 TI - More about Helicobacter pylori, antibiotics and IBD. PMID- 15361089 TI - Association between Helicobacter pylori Infection in gastric cancer, ulcers and gastritis in Iranian patients. PMID- 15361091 TI - Pharmacologic management and treatment of gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) is one of the most common diagnoses in daily practice. Diagnosis can be made on symptom evaluation, on pH-monitoring or on endoscopic findings. In contrast to commonly held opinion there is no strong evidence that lifestyle factors are a dominant factor in the pathophysiology of GERD. The various agents currently used for treatment of GERD include mucoprotective substances, antacids, H(2)-blockers and proton pump inhibitors. This article gives an overview of the pharmacological management of GERD and focuses on the differential therapy of endoscopy-negative GERD, GERD with esophagitis and maintenance therapy. PMID- 15361092 TI - 5-Aminolaevulinic acid-induced photodynamic therapy and photodetection in Barrett's esophagus. AB - Barrett's esophagus is a precursor of adenocarcinoma of the esophagus. This cancer has the fastest growing incidence of any solid tumor in the Western world. Surveillance of Barrett's esophagus is routinely undertaken to detect early malignant transformation. However, ablative endoscopic treatments are available and these can obliterate the abnormal epithelium, allowing neosquamous regrowth. Photodynamic therapy using 5-aminolaevulinic acid (ALA) is such a technique. In this non-thermal method of ablation, ALA is metabolized to produce the photosensitizer protoprophyrin IX. This, together with light and oxygen, produces local tissue destruction. Fluorescence detection using ALA has also been used to identify areas of dysplasia and thus enhance positive biopsy yield. The use of ALA in photodynamic therapy and photodetection is reviewed. PMID- 15361093 TI - Efficacy of botulinum toxin injection before pneumatic dilatation in patients with idiopathic achalasia. AB - Graded pneumatic dilatation (PD) is an appropriate long-term therapy and botulinum toxin injection (BT) is a relatively short-term therapy in idiopathic achalasia. Their combination has not been previously scrutinized. This study aimed to evaluate the role of BT in enhancing the efficacy of PD with 30 mm balloons. Patients who underwent PD with 30 mm balloons after botulinum toxin injections and a group of age- and sex-matched controls who were treated only with PD were enrolled in the study. Symptom scores were taken before, 1 month after and then every 3 months after PD. There were no significant differences between the two groups in gender, duration or severity of symptoms. One of the 12 patients in the case group relapsed 30 months after PD but the others were in remission for an average of 25.6 months. In the control group, all the patients relapsed after a mean of 12.6 months and needed a 35-mm PD. The cumulative remission rate was significantly higher in the case group compared with the control group (P < 0.01). The mean symptom score decreased by 76% in the case group (P < 0.001) and 53% in the controls (P < 0.01) at the end of the first month. Neither age, sex, nor duration or severity of symptoms were predictive of patients' responses to treatment. It seems that BT may be a meaningful enhancing factor in long-term efficacy of PD. PD with a 30 mm balloon after a BT session may resolve the need for the future higher grade PD. PMID- 15361094 TI - Use of radioisotopic esophageal transit in the assessment of patients with symptoms of reflux and non-specific esophageal motor disorders. AB - The purposes of this study were to assess the esophageal clearance of a radioisotopic bolus in patients with symptoms of reflux and evaluate the impact of manometric abnormalities on scintigraphic esophageal transit. Esophageal clearance was assessed in a supine position and indicated by the retained radioactivity in the esophagus at 10, 20, 30 and 40 s after the ingestion of a liquid bolus labeled with 2 mCi 99 mTc-SC. The study included 214 consecutive patients with symptoms of reflux and 11 normal controls. The results were compared to the motility findings detected on manometry performed on a separate occasion. Esophageal manometry was normal in 93 patients. Nonspecific esophageal motor disorders were identified in 121 patients and were classified into: 'predominantly nonpropagated activity', 'predominantly low-amplitude peristaltic contractions' and 'miscellaneous disorders' diagnosed in 27, 47 and 47 patients, respectively. The radionuclide clearance was significantly delayed in the overall group of patients compared with that of normal controls (P < 0.001); in patients with reflux symptoms and nonspecific esophageal motor disorders compared with patients with reflux symptoms and 'normal manometry' (P < 0.01 at 20 s); and in patients with reflux symptoms and 'normal manometry' compared with the control group (P < 0.01 at 20 s). Abnormal radioisotope clearances were detected in 88% of patients with 'predominantly nonpropagated activity', in 70% of patients with 'predominantly low-amplitude peristaltic contractions' and in 57% of patients with 'miscellaneous disorders'. Radioisotopic esophageal clearance abnormalities are frequently observed in patients with reflux symptoms and are more likely to be associated to hypomotility disorders, i.e. nonpropagated motor activity or low amplitude contractions. PMID- 15361095 TI - Characteristics of patients with erosive and nonerosive GERD in high-Helicobacter pylori prevalence region. AB - It is still not known whether there are differences between erosive and nonerosive GERD. The aim of the present study is to evaluate the prevalence of Helicobacter pylori (HP) infection, and other differences between erosive and nonerosive gastroesophageal reflux disease (NERD) patients. One-hundred and four consecutive GERD patients (mean age: 41.6 +/- 12.3 years) were interviewed, endoscoped and tested for HP. Erosive GERD was defined according to the Los Angeles classification. Patients who had no erosions in the esophagus but complained of heartburn or/and acid regurgitation at least twice a week and for whom these symptoms had a negative impact on daily activities were considered to be NERD patients. Erosive GERD was identified in 53 (51%) patients (mean age: 41.0 +/- 12.7 years) and NERD in 51 (49.0%) patients (mean age: 42.2 +/- 11.9 years). HP infection was found in 32 (60.4%) erosive GERD patients, and 41 (80.4%) NERD patients, P < 0.05. Multivariate analysis revealed that there were two statistically significant prediction factors for NERD: female sex with odds ratio (OR) of 6.34 (95% CI: 2.41-16.64; P = 0.0002) and HP infection with odds ratio (OR) of 3.28 (95% CI: 1.26-8.58; P = 0.015). The presence of HP and female sex are found to be statistically significant predictors of NERD. PMID- 15361097 TI - Histological inflammatory changes after surgery at the epithelium of the distal esophagus in patients with Barrett's esophagus: a comparison of two surgical procedures. AB - There are many reports concerning the surgical treatment of patients with Barrett's esophagus, but very few focus on histological changes of inflammatory cells in squamous and columnar epithelium before and late after classic antireflux or acid suppression-duodenal diversion surgery. We evaluate the impact of these procedures in the presence of intestinal metaplasia, dysplasia and Helicobacter pylori in the columnar epithelium. Two groups of patients were studied, 37 subjected to classic antireflux and 96 to acid suppression-duodenal diversion operations. They were subjected to endoscopic and histological studies before and at 1, 3 and more than 5 years after surgery. Manometric evaluations and 24 h pH monitoring were performed before and at 1 year after surgery. The presence of inflammatory cells at both the squamous and columnar epithelium was significantly higher at the late follow up in patients subjected to classic antireflux surgery compared with patients subjected to acid suppression-duodenal diversion operations (P < 0.02 and P < 0.001, respectively). Intestinal metaplasia, present in 100% of patients before surgery, had decreased significantly at 3 years after surgery in patients subjected to acid suppression duodenal diversion operations compared with classic antireflux procedures, 75% versus 53%, respectively (P < 0.001). The presence of Helicobacter pylori did not vary before or after surgery in either group. In conclusion, acid suppression duodenal diversion operations are followed by a decreased presence of inflammatory cells in both squamous and columnar epithelium compared with classic antireflux surgery in patients with Barrett's esophagus. Intestinal metaplasia and dysplasia and inflammation findings were also less common after acid suppression-duodenal diversion operation. PMID- 15361096 TI - Psychometric and methodological characteristics of a culturally adjusted gastroesophageal reflux disease questionnaire. AB - Population-based data regarding the prevalence of gastroesophageal reflux disease (GERD) in Turkey is lacking. Therefore, a valid and reliable instrument for population-based study of GERD is important. The purpose of the study is to establish the feasibility, reproducibility, reliability and validity of the Turkish version of the GERD questionnaire. The study sample of 630 people was randomly recruited from the 20- to 65-year-old-population of Menderes, Izmir. Face to face interviews were conducted using the Turkish GERD questionnaire. The questionnaire was translated into Turkish, and then test-retest reliability of the study was carried out with 25 individuals. Internal reliability was measured using Cronbach's alpha. Additionally, a validity study was performed at the gastroenterology outpatient clinic at Ege University, Faculty of Medicine (45 people). This Turkish GERD questionnaire was easily understood and completed in a reasonable time by members of the population for whom the survey was intended. For all questions (omitting the demographic questions) kappa statistics were calculated; the median kappa for the outpatient assessment was 0.82. The sensitivity of the questionnaire concerning heartburn was strong and the specificity of the test was weak. By calculating Cronbach's alpha coefficient, we estimated the internal consistency or the reliability of each component. The Cronbach's alpha coefficients were above 0.70. This instrument can be used for large-scale population based studies in Turkey. PMID- 15361098 TI - Endoscopic ablation of Barrett's esophagus using argon plasma coagulation: a prospective study after fundoplication. AB - The aim of the prospective clinical study presented here is to test the effectiveness of a multimode approach consisting of argon plasma coagulation combined with laparoscopic fundoplication in the management of Barrett's esophagus. Argon plasma coagulation was performed in 19 patients with Barrett's esophagus who had previously undergone surgical antireflux treatment. The mean follow-up time was 17 months, ranging between 6 and 27 months. Squamous epithelium was completely restored in all patients. In 68.4% of cases two sessions were required. The most frequent complications were chest discomfort and retrosternal pain. In 11 patients the symptoms lasted 3 days and in six cases persisted for a longer period, requiring analgesic medication. Short-term dysphagia and odynophagia were observed in four patients. PMID- 15361099 TI - Phase I study of the combination of nedaplatin, adriamycin and 5-fluorouracil for treatment of advanced esophageal cancer. AB - This trial was conducted to determine the maximum-tolerated dose, principal toxicity, and recommended dose (RD) for the phase II study of the combination of nedaplatin (NED), adriamycin (ADM), and 5-fluorouracil (5-FU) in patients with advanced esophageal cancer. Patients with previously untreated esophageal cancer were eligible if they had performance status 0-1, were 75 years or younger and had adequate organ function. The dose of NED, the key anticancer platinum complex drug, was increased from 60 to 70, and 80 mg/m(2) on day 1. ADM and 5-FU were administered at fixed doses (30 mg/m(2) on day 1, and 700 mg/m(2) on days 1-5). The dose-limiting toxicities of NED were neutropenia and severe diarrhea, and its maximum-tolerated dose and RD were 70 mg/m(2) and 60 mg/m(2), respectively. There were four responders among the six patients administered the RD. The present study thus revealed combination chemotherapy with NED, ADM, and 5-FU to be active and well-tolerated and to warrant phase II study. PMID- 15361100 TI - Telomerase activity and survival of late-stage South African esophageal carcinoma patients. AB - Squamous cell carcinoma of the esophagus is a cancer with a high incidence in South Africa. We have investigated the prognostic value of telomerase activity in tumors as well as nearby normal tissue. Biopsies from 98 patients (71 men and 27 women) were analyzed using an adaptation of the TRAP assay. We found all tumor biopsies to have moderate to high telomerase activity, while one third of biopsies from normal mucosa were negative. The telomerase activity level of the tumors had no prognostic value (P = 0.95) as determined by the log rank test. A P value of 0.02 was found when the telomerase-negative and moderately positive normal biopsies were grouped together and compared to those with high activity. Our results show that telomerase activity of normal mucosa in the vicinity of the tumor can identify a population of patients with significantly worse prognosis, even in late stage patients. PMID- 15361101 TI - Constituents of areca chewing related to esophageal cancer risk in Taiwanese men. AB - Two most common types of areca chewing are noted in Taiwan: raw betel fruit with Piper betle inflorescence or folded in betel leaf. Piper betle inflorescence contains carcinogens, whereas betel leaf includes anticarcinogenic agents. One hundred and twenty-six esophageal squamous-cell-carcinoma patients and 279 healthy controls, all men, were analyzed. Areca chewers were 4.4 times (95% CI, 2.2-8.8) more likely to develop esophageal cancer than non-chewers. Sixty-five of the patients were areca chewers, of which, 61 (93.9%) chewed areca with Piper betle inflorescence, none chewed it with betel leaf and four (6.1%) chewed both. Of the 24 controls who were chewers, 10 (41.7%), three (12.5%) and 11 (45.8%) chewed areca with Piper betle inflorescence, betel leaf, and both, respectively. Multivariate analysis showed that subjects who chewed areca with Piper betle inflorescence were 24.4 times (95% CI 3.9-154.4) more likely to develop esophageal cancer than those who chewed areca with betel leaf or with both leaf and inflorescence. Our epidemiologic findings suggest parts of the same Piper plant contains carcinogenic and anticarcinogenic substances. PMID- 15361102 TI - Chemoradiation therapy is effective for the palliative treatment of malignant dysphagia. AB - Between 1993 and 2001, 106 patients with esophageal cancer were reviewed at a multidisciplinary clinic and treated with palliative intent by chemoradiation therapy. This study assesses the palliative benefit on dysphagia and documents the toxicity of this treatment. The study population comprised 72 men and 34 women with a median age of 69 years. Patients were treated with a median radiation dose of 35 Gy in 15 fractions with a concurrent single course of 5 FU based chemotherapy. Dysphagia was measured at the beginning and completion of treatment and at monthly intervals until death, using a modified DeMeester (4 point) score. Treatment was well tolerated, with only 5% of patients failing to complete therapy. The treatment-related mortality was 6%. The median survival for the study population was 7 months. The median baseline score at presentation was 2 (difficulty with soft food). Following treatment, 49% of patients were assessed as having a dysphagia score of 0 (no dysphagia). Seventy-eight per cent had an improvement of at least one grade in their dysphagia score after treatment. Only 14% of patients showed no improvement with treatment. Fifty-one per cent maintained improved swallowing until the time of last follow-up or death. This single-institution study shows that chemoradiation therapy administered for the palliation of malignant dysphagia is well tolerated and produces a sustainable normalization in swallowing for almost half of all patients. PMID- 15361103 TI - Tracheoesophageal fistula secondary to chemotherapy for malignant B-cell lymphoma of the thyroid: successful surgical treatment with jejunal interposition and mesenteric patch. AB - We report a case of tracheoesophageal fistula (TEF) secondary to chemotherapy for primary thyroid lymphoma. A 65-year-old man with a short history of a rapidly enlarging neck mass was diagnosed as having thyroid lymphoma of diffuse, large B cell type. The TEF occurred during the first course of chemotherapy including cyclophosphamide, doxorubicin, vincristine and prednisolone. After placing a feeding gastrostomy without oral intake, eight cycles of chemotherapy were completed and complete remission was achieved. Although the cervical mass disappeared, TEF and esophageal stenosis persisted. Total thyroidectomy and resection of the stenotic cervical esophagus were carried out followed by interposition of the revascularized jejunum and its mesenteric patch to cover the TEF. This seems to be the first report of a TEF caused by chemotherapy for primary thyroid B-cell lymphoma. A variety of treatments for TEF including simple closure, tracheal resection, colonic bypass and muscle flap have been reported with low success rates. Our procedure using a jejunal mesenteric patch seems to be unique and may be a new treatment strategy for TEF. PMID- 15361104 TI - Esophagectomy in a patient with AIDS. AB - As the life expectancy of AIDS patients continues to increase due to improved anti-retroviral therapy less of these patients die of HIV-related illnesses. Dysphagia is a common complaint in AIDS patients and usually results from a fungal esophagitis. While cancer of the esophagus is occasionally found in AIDS patients, we report our experience with an AIDS patient diagnosed with a squamous cell esophageal malignancy who received pre-operative radiation and chemotherapy, followed by transhiatal esophagectomy. The patient is alive 16 months post operatively. The transition of HIV/AIDS from an acute fulminant disease to a chronic condition mandates that these patients should receive full and standard therapy for their esophageal malignancies. PMID- 15361105 TI - The north-south divide in health research publishing: what can smaller journals do? PMID- 15361106 TI - Stage-specific effects of host plasma factors on the early sporogony of autologous Plasmodium falciparum isolates within Anopheles gambiae. AB - Summary Quantitatively assessing the impact of naturally occurring transmission blocking (TB) immunity on malaria parasite sporogonic development may provide a useful interpretation of the underlying mechanisms. Here, we compare the effects of plasma derived from 23 naturally infected gametocyte carriers (OWN) with plasma from donors without previous malaria exposure (AB) on the early sporogonic development of Plasmodium falciparum in Anopheles gambiae. Reduced parasite development efficiency was associated with mosquitoes taking a blood meal mixed with the gametocyte carriers' own plasma, whereas replacing autologous plasma with non-immune resulted in the highest level of parasite survival. Seven days after an infective blood meal, 39.1% of the gametocyte carriers' plasma tested showed TB activity as only a few macrogametocytes ingested along with immune plasma ended up as ookinetes but subsequent development was blocked in the presence of immune plasma. In other experiments (60.9%), the effective number of parasites declined dramatically from one developmental stage to the next, and resulted in an infection rate that was two-fold lower in OWN than in AB infection group. These findings are in agreement with those in other reports and go further by quantitatively examining at which transition stages TB immunity exerts its action. The transitions from macrogametocytes to gamete/zygote and from gamete/zygote to ookinete were identified as main targets. However, the net contribution of host plasma factors to these interstage parasite reductions was low (5-20%), suggesting that irrespective of the host plasma factors, mosquito factors might also lower the survival level of parasites during the early sporogonic development. PMID- 15361107 TI - Submicroscopic Plasmodium falciparum infections and multiplicity of infection in matched peripheral, placental and umbilical cord blood samples from Gabonese women. AB - Summary In malaria-endemic regions, pregnant women are more susceptible to malarial infections than non-pregnant women. The main objective of this study, which was conducted in the malaria hyperendemic town of Lambarene (Gabon, Central Africa), was to characterize Plasmodium falciparum infections in peripheral, placental and cord blood from women of different gravidities with submicroscopic infections. Using the P. falciparum merozoite surface protein 2 (MSP 2)* gene as a polymorphic marker in polymerase chain reactions, we analysed genetic diversity and multiplicity of infection in isolates from all three kinds of samples of 184 pregnant women at delivery. We detected infection in 44% of the women who were originally negative by microscopy. Equally important was the finding that the placenta had the highest prevalence of infection (P < 0.001). There was no correlation with gravidity status or age of the patients. The multiplicities of infection in the peripheral and placental blood samples did not differ and single infection was observed in cord blood, independently of the gravidity. The FC27/MSP 2 was the predominant allelic family. The major FC27 alleles detected in the peripheral, placental and cord blood were sequenced and found to be closely related to the published K1 form sequence. Below microscopy level, the placenta remains the most infected organ and this submicroscopic carriage of parasites may contribute to the development and maintenance of immunity to malaria during pregnancy. PMID- 15361108 TI - Artemisinin-based combination therapy reduces expenditure on malaria treatment in KwaZulu Natal, South Africa. AB - INTRODUCTION: There is growing international evidence that artemisinin-based combination therapy (ACT) is one of the few effective measures available to 'Roll Back Malaria'. However, concerns about the costs and affordability of ACT are obstacles to its widespread implementation. This paper explores some economic aspects of the implementation of artemether-lumefantrine (AL) to replace sulphadoxine-pyrimethamine (SP) in the KwaZulu Natal (KZN) province, South Africa. METHODS: Recurrent and capital costs for malaria treatment were compared at baseline and post-intervention for nine clinics and a sentinel rural district hospital. Changes in the unit costs of, and total expenditure on, malaria services were calculated and the cost effectiveness of AL relative to SP was assessed. RESULTS: The number of outpatient malaria cases and inpatient admissions both declined by 94% between 2000 and 2002. After accounting for the role of concurrent improvements in vector control, it was conservatively estimated that 36% of the decline in outpatient cases and 46% for inpatient admissions was attributable to changing the first-line drug to AL. Although AL is considerably more expensive than SP, its improved cure rate and reduced malaria transmission resulted in an estimated 201,065 US dollars cost saving in 2002 alone for the subdistrict studied. DISCUSSION: In the context of effective vector control and low efficacy of existing monotherapy, ACT can reduce total expenditure on malaria services. However, the relevance of these findings requires careful consideration in countries with currently effective treatment policies and higher intensity malaria transmission. PMID- 15361109 TI - The difference between effectiveness and efficacy of antimalarial drugs in Kenya. AB - OBJECTIVE: To demonstrate the difference between effectiveness and efficacy of antimalarial (AM) drugs in Kenya. METHODS: We undertook a series of linked surveys in four districts of Kenya between 2001 and 2002 on (i) community usage of nationally recommended first- and second-line AM drugs; (ii) commonly stocked AM products in the retail and wholesale sectors; and (iii) quality of the most commonly available first- and second-line AM products. These were combined with estimates of adherence and clinical efficacy to derive overall drug effectiveness. RESULTS: The overall modelled effectiveness for sulphadoxine pyrimethamine (SP) was estimated to be 62% compared with 85% for reported SP clinical efficacy. For amodiaquine the modelled effectiveness was 48% compared with 99% reported efficacy during the same time period. CONCLUSIONS: The quality of AM products and patient adherence to dosage regimens are important determinants of drug effectiveness, and should be measured alongside clinical efficacy. Post-registration measures to regulate drug quality and improve patient adherence would contribute significantly to AM drug performance. PMID- 15361110 TI - Efficacy of chloroquine, sulphadoxine-pyrimethamine and amodiaquine for treatment of uncomplicated Plasmodium falciparum malaria in Kajo Keji county, Sudan. AB - To provide advice on the rational use of antimalarial drugs, Medecins Sans Frontieres conducted a randomized, an open label efficacy study in Kajo Keji, an area of high transmission of malaria in southern Sudan. The efficacy of chloroquine (CQ), sulphadoxine-pyrimethamine (SP) and amodiaquine (AQ) were measured in a 28-day in vivo study, with results corrected by PCR genotyping. Of 2010 children screened, 115 children aged 6-59 months with uncomplicated Plasmodium falciparum malaria were randomized into each group to receive a supervised course of treatment. Of these, 114, 103 and 111 were analysed in the CQ, SP and AQ groups, respectively. The overall parasitological failure rates at day 28 were 93.9% [95% confidence interval (CI) 87.3-97.3] for CQ, 69.9% (95% CI 60.0-78.3) for SP, and 25.2% (95% CI 17.7-34.5) for AQ. These results provide important missing data on antimalarial drug efficacy in southern Sudan. They indicate that none of the drugs could be used in monotherapy and suggest that even in combination with artemisinin, cure rates might not be efficacious enough. We recommend a combination of artemether and lumefantrine as first-line treatment for uncomplicated P. falciparum malaria cases in Kajo Keji county. PMID- 15361111 TI - Anti-leishmanial activity of a new formulation of amphotericin B. AB - The effectiveness of albumin microspheres loaded with amphotericin B was tested in an in vivo model of visceral leishmaniasis using the golden hamster. Free and encapsulated amphotericin B was tested at the dose of 1 mg/kg given by the intracardiac route on days 25, 26 and 27 post-infection (p.i.) to treat animals previously infected with 10(7) stationary promastigotes by the intracardiac route. Encapsulated amphotericin was highly effective against infection causing a reduction of 88.8% and 87.2% in the early stage of infection (day 32 p.i.) and of 66.7% and 54% in a later stage of infection (day 135 p.i.) in liver and spleen parasite load respectively, compared with untreated animals, whereas free amphotericin was inactive. Lymphocyte proliferation was restored together with an increase in CD4(+) subsets in animals treated with encapsulated amphotericin B, but not in those treated with the non-encapsulated compound. Antibody responses did not increase after treatment with encapsulated amphotericin B with antibody levels remaining at base levels for most animals in contrast to those of untreated or treated with free amphotericin, where in most animals the antibody levels sharply increased. This new formulation could be a more economical alternative to liposomes for the treatment of visceral leishmaniasis with amphotericin B. PMID- 15361112 TI - Applicability of a monoclonal antibody-based dipstick in diagnosis of urinary schistosomiasis in the Central Region of Ghana. AB - Summary We tested a rapid visually read monoclonal antibody (MoAb) based dipstick assay for specific diagnosis of urinary schistosomiasis against microscopy and the use of haematuria and proteinuria in a schistosomiasis haematobia endemic area in the Central Region of Ghana. The study group consisted of 141 school children (83 males, 58 females) aged 8-19 years. A total of 129 of 141 (91.5%) submitted stool samples, and 7.8% had Schistosoma mansoni, 55% had hookworms and 6.2% had tapeworms. The presence of S. mansoni and intestinal parasites did not appear to influence the results of the MoAb-dipstick assay. The urinary schistosomiasis prevalence by MoAb-dipstick (78%) was higher (P < 0.05) than the estimate by microscopy (60.3%), microhaematuria (27%) and proteinuria (30.5%). The MoAb-dipstick correctly identified 98.8% of microscopically confirmed cases and missed one (1.3%). The dipstick was also positive for 26 of 56 (46.4%) egg negative individuals, thereby giving a sensitivity of 98.8% and a specificity of 53.6%. On the other hand, microhaematuria and proteinuria were 38.8% and 30.6% sensitive, and 91.1% and 69.6% specific, respectively. Microhaematuria and proteinuria were less sensitive (P < 0.05) than both microscopy and MoAb dipstick. PMID- 15361113 TI - Environmental control of schistosomiasis through community participation in a Moroccan oasis. AB - Summary Akka oasis, in the province of Tata, southern Morocco, is one of the oldest foci of urinary schistosomiasis in Morocco where transmission is still taking place. We report the results of two studies: a cross-sectional snail survey investigated the distribution of Bulinus truncatus in relation to habitat factors in the Akka traditional irrigation system. The presence of aquatic vegetation, especially Potamogeton sp. was identified as a key factor determining snail occurrence and abundance in canals, impoundments and isolated small puddles and streamlets in the Akka riverbed. In a participatory rapid appraisal, the community identified snail control as a way to reduce transmission of schistosomiasis. Without any further outside incentives, the local irrigation committee implemented repeated cleaning and vegetation removal in canals. A longitudinal study evaluated the effect of these measures on populations of B. truncatus. Snail and egg mass densities showed significant reductions after repeated vegetation clearing in the study sites. The participatory approach led to low-cost, environment-friendly schistosomiasis control measures that were effective and sustainable. PMID- 15361114 TI - Childhood malignancies in French Polynesia during the 1985-1995 period. AB - We report an estimation of the incidence of childhood cancer among natives of French Polynesia (FP) during the 1985-1995 period. Our data were acquired from the Cancer Registry of FP and through an extensive investigation of other potential sources of information. The mean population of children between 1985 and 1995 was estimated to be 63 401 inhabitants, 32 487 of whom were boys and 30 914 girls, born and residing in FP. During the 1985-1995 period, 87 incident cases of childhood cancer were recorded among inhabitants born in FP or of an unknown place of birth (n = 2). Childhood cancer incidence had attained 125 cases/million child years and was very similar among girls (126 x 10(-6)) and boys (123 x 10(-6)), this incidence being slightly lower than among other populations of similar ethnic origin: Standardized Incidence Ratio (SIR) = 0.8 (95% CI: 0.7-1.0) when compared with New Zealand Maoris and SIR = 0.8 (95% CI: 0.6-1.0) when compared with natives from Hawaii. For both sexes considered together, the most frequent cancer type was leukaemia, followed by central nervous system (CNS) malignancies, neuroblastoma, and non-Hodgkin's lymphoma (NHL). Only one case of gonadal and germ cell tumours and one case of carcinoma were reported. Childhood cancer incidence was predominant among children living in the Windward, Leeward and Marquesas Islands and the Tuamotu-Gambier archipelago, but lower in the Austral Islands. The incidence of acute non lymphocytic leukaemia (ANLL) decreased from 3.3 x 10(-5) between 1985 and 1989, an unexpectedly high incidence, to 0.8 x 10(-5) between 1990 and 1995. PMID- 15361115 TI - Soil-transmitted helminthiasis in Myanmar and approximate costs for countrywide control. AB - Previous surveys in Myanmar indicate that soil-transmitted helminths (STH) are a significant public health problem. However the majority of these surveys are small scale or hospital based. To date there has been no countrywide assessment of the prevalence and intensity of STH infection. We conducted a survey (June 2002-June 2003) to assess the magnitude of STH infection in school age children in each of the different ecological areas of the country. In each area, five schools were selected and from each one, 50 children were randomly chosen. Parasitological and nutritional data were collected from each child. We found an overall prevalence of infection of 69%: 18% had a moderate-heavy intensity of infection, 22% were anaemic. A pilot exercise to treat 25,000 school age children in the 200 schools of Nyangdone Township tested the strategy before scaling up and to generate some rough costings. A crude calculation of the expenditure to buy and deliver the drugs, including the costs incurred in the teacher training, concluded that the cost per child was approximately 0.05 USD. PMID- 15361116 TI - An outbreak of leptospirosis in Orissa, India: the importance of surveillance. AB - OBJECTIVE: To demonstrate the importance of surveillance systems in detecting emerging diseases and highlighting the strengths and weaknesses of an existing one. METHODS: The Orissa multi-disease surveillance system (OMDSS) was introduced in November 1999. Reporting units from the periphery send data to the district on a weekly basis. These reports are analysed regularly. A district task force (DTF) was available to intervene in the event of an outbreak. The OMDSS detected an increasing number of cases with fever and jaundice in June 2002. The DTF investigated this outbreak using clinical, epidemiological and laboratory tools to identify its cause. RESULTS: This outbreak, in a remote corner of India, was detected within 4 days by an existing surveillance system. Action was initiated within 24 h, but it took approximately two more weeks for the causative agent to be diagnosed. A total of 143 people were suspected to have leptospirosis between 23 June 2002 and 31 July 2002. The attack rate was 5.95% and the case fatality ratio (CFR) was 7.69%, both lower than outbreaks reported elsewhere in India. While males were infected more often than females, the CFR was higher among females and among the 6-15 year age groups. Exposure to infected water in a canal was the probable cause of the outbreak. IgM antibodies were positive in 33 of the patients and six patients tested positive for PCR and culture. Leptospirosis interrogans serovar canicola, Leptospirosis interrogans serovar pomona and Leptospirosis interrogans serovar hebdomadis were isolated. CONCLUSIONS: Leptospirosis is a new disease in this region of India. This outbreak was detected and diagnosed because of the surveillance system. The prompt response helped in containing the outbreak early enough. However, the morbidity and mortality could have been further mitigated if the delays in transmitting information had been minimized. An adequate laboratory support would have also helped considerably. We conclude stressing the importance of surveillance as a public health tool. PMID- 15361117 TI - Risk factors and clinical features associated with severe dengue infection in adults and children during the 2001 epidemic in Chonburi, Thailand. AB - OBJECTIVES: Dengue haemorrhagic fever (DHF) is an important cause of morbidity in South-east Asia and used to occur almost exclusively in young children. In recent years, there has been a progressive shift in age-distribution towards older children and adults. We investigated an outbreak in 2001 in both children and adults, in an endemic area of Thailand. METHODS: Retrospective study of 347 patients with serologically confirmed dengue infection admitted to Chonburi Hospital during an epidemic in 2001. RESULTS: A total of 128 (37%) patients had dengue fever (DF) and 219 (63%) had DHF. Patients with DHF were significantly older than patients with DF (11 years vs. 8 years). Clinical bleeding was noted in 124 individuals, both with DF (n = 24) and DHF (n = 100), and significantly more frequently in adults. Twenty-nine (13.2%) of all DHF cases were caused by primary infection. Secondary dengue infection was associated significantly with the development of DHF in children, OR (95% CI) = 3.63 (1.94-6.82), P < 0.0001, but not in adults, OR (95% CI) = 0.6 (0.02-6.04), P = 1. Unusual clinical manifestations were observed in 23 patients: three presented with encephalopathy and 20 with highly elevated liver-enzymes. In the latter group, four patients were icteric and nine had gastrointestinal bleeding. CONCLUSION: These results indicate that DHF in South-east Asia is common in both children and adults. In dengue-endemic countries, dengue should be considered as a differential diagnosis in patients with clinical gastrointestinal bleeding in association with increased liver enzymes. PMID- 15361118 TI - Effectiveness of two annual, single-dose mass drug administrations of diethylcarbamazine alone or in combination with albendazole on soil-transmitted helminthiasis in filariasis elimination programme. AB - A longitudinal community-trial on the control of soil-transmitted helminths (STHs), as part of a lymphatic filariasis elimination campaign, was taken up in two revenue blocks of southern India in the years 2001 and 2002 to assess the impact of two annual single-dose mass drug administration (MDA) of diethylcarbamazine (DEC) + albendazole (ALB) with that of DEC alone. The prevalences and intensities of STHs were studied among cross-sectional samples of school children aged 9-10 years by using the Kato-Katz technique at baseline and 11 months after each MDA. The combined drug mass treatment produced a higher reduction in the prevalence (RIP) (51-77%) and the egg reduction rate (ERR) (92 98%) compared with 12-15% RIP and 58-62% ERR of DEC alone mass treatment. The effect of two-drug therapy after two annual treatments was relatively long lasting as shown by RIP and ERR indicating that the reinfection rates were relatively lower in this approach than single-drug therapy. This study demonstrates that mass drug co-administration of DEC + ALB in Global Programme for Elimination of Lymphatic Filariasis (GPELF) targeted at the community had a synergistic and sustainable effect against soil-transmitted helminthiasis and provided considerable 'beyond filariasis' benefits. The additional advantages accrued to the community underscore the importance of scaling-up GPELF to cover the entire population at risk. PMID- 15361119 TI - Helicobacter pylori antigens in the faeces of asymptomatic children in the Buea and Limbe health districts of Cameroon: a pilot study. AB - OBJECTIVE: To determine the prevalence and identify intra-familial risk factors associated with Helicobacter pylori infection in a paediatric population. METHODS: Cross-sectional study in the Buea and Limbe health districts, South West Cameroon. Stool samples were collected from 176 randomly selected apparently healthy children from two communities with different socioeconomic status. They comprised 86 males and 90 females aged 0-10 years with a mean age of 4.29. Helicobacter pylori status was determined using an enzyme-linked immunosorbent assay, the H. pylori stool antigen (HpSA) test. The test uses polyclonal anti-H. pylori capture antibody to detect H. pylori antigens in human stool. Epidemiological data were analysed using the Fisher test and odds ratio (OR) at 95% confidence intervals (CI). RESULTS: The overall prevalence of H. pylori was 52.27% (92 of 176). Univariate analysis showed that H. pylori prevalence was significantly higher in children of the low socioeconomic class, 62.50% (55 of 88) than in those of the high socioeconomic class, 42.05% (37 of 88) (P < 0.05; OR = 2.41, 95% CI: 1.26-4.64). Helicobacter pylori prevalence increased with age from 37.50% (12 of 32) for children aged <3 years, 50.00% (53 of 106) aged 3-6 years and 71.05% (27 of 38) aged 7-10 years (P > 0.05; OR = 0.81, 95% CI: 0.34 1.91). The frequency of infection was significantly higher in males, 64.00% (55 of 86) than in females, 41.11% (37 of 90), (P < 0.05; OR = 2.67, 95% CI: 1.39 5.17). CONCLUSIONS: This study highlights the continuing importance of age, sex and socioeconomic status in the acquisition of H. pylori infection. PMID- 15361121 TI - The heat-shock protein receptors: some answers and more questions. AB - The existence of heat-shock protein (HSP) receptors on antigen-presenting cells (APCs) was hypothesized in 1994. The first such receptor, CD91 or LRP, was identified and characterized in 2000. The pace of attribution has quickened since and during the last three years alone, six putative HSP receptors have been identified. These include CD40, LOX-1, CD36, Toll-like receptor-2 (TLR-2), TLR-4 and SR-A. The literature on HSP receptors on APCs is critically examined in this review and future directions are imagined. PMID- 15361123 TI - Development of a multiplex PCR-SSP method for Killer-cell immunoglobulin-like receptor genotyping. AB - Killer-cell immunoglobulin-like receptors (KIRs) on natural killer (NK) cells recognize groups of HLA class I alleles. Recent work suggests that KIR genotype may affect the outcome of hematopoietic stem-cell transplants and that prospective KIR typing maybe of benefit in future matching of donors and recipients. A simple and informative KIR genotyping method was developed using a multiplex polymerase chain reaction-sequence-specific primer strategy. This method contains four multiplex reactions for detecting all functional KIR genes, including some 2DS4 variants that harbor a common deletion. Primer pairs were designed to provide short amplicons (108-565 bp) that can be analyzed by agarose gel electrophoreses or by automated electrophoretic systems. This method was evaluated in a blinded survey with the NK/KIR Phase II QC Panel (a total of 16 cell lines) from the 14th International Histocompatibility Workshop (IHWS), and the results are 100% concordant with the consensus genotype. Results in further KIR genotyping of 20 reference cell lines from the 10th IHWS were consistent with previously published genotypes, matching those of one study in instances where different genotypes have been previously reported. The genotypes obtained in this study may be helpful to other labs developing KIR genotyping methods in resolving typing discrepancies and in detecting common deletion variants of 2DS4. This method can save labor and reagent costs. It provides good results from partially degraded template DNA due to short amplicons in this method. It is convenient to use in both clinical and research laboratories. PMID- 15361122 TI - High level of chromosome 15 aneuploidy in head and neck squamous cell carcinoma lesions identified by FISH analysis: limited value of beta2-microglobulin LOH analysis. AB - In cancer research, loss of heterozygosity (LOH), defined by microsatellite markers, is frequently used in the identification of gene loss. Especially, genomic alterations in the human leukocyte antigen (HLA) genes and the beta2 microglobulin (beta2m) gene on chromosome 15 are of interest regarding their function in the immune system. Because LOH analysis detects any allelic imbalance and not just allelic loss, we evaluated the LOH analysis in 11 head and neck squamous cell carcinoma (HNSCC) lesions using fluorescence in situ hybridization (FISH). The 11 tumors were selected out of 53 HNSCC lesions based upon beta2m LOH analysis and beta2m expression. Centromere 1 and 15 FISH were developed to determine the chromosome 15 copy number. Sequence-based mutation analysis of beta2m was conducted on tumors without beta2m expression; no mutations in the coding sequences were found. For five HNSCC lesions with LOH and beta2m expression, centromere 15 FISH indicated gain rather than loss. In the majority of the 11 HNSCC lesions, FISH showed centromere 1 and 15 heterogeneity throughout the tumor. Moreover, FISH indicated a more complex chromosome 1 and 15 distribution than could be concluded from microsatellite LOH analysis. Our results show that microsatellite LOH analysis does not represent the beta2m gene copy number and support the results obtained from comparative genomic hybridization (CGH) studies. Conclusions on genomic alterations in tumors cannot be based on LOH data only but depend on the results of immunohistochemical staining, FISH, and CGH. PMID- 15361124 TI - TNF-alpha-308A allele, a possible severity risk factor of hemorrhagic manifestation in dengue fever patients. AB - Among the several hypothesis postulated to explain the pathogenesis of severe dengue disease, the model of immunopathogenesis is the most supported one with a likely important role played by the cascade of cytokines. This work describes single-nucleotide polymorphism of tumor necrosis factor (TNF)-alpha, interferon gamma, interleukin (IL)-6, transforming growth factor-beta1, and IL-10 in patients with dengue virus infections and analyzes their relation with clinical manifestations of the disease. Because cytokine gene polymorphisms affect cytokine production, the significant increase of the TNF-308A allele we have observed among patients with dengue fever (DF) with hemorrhagic manifestations compared to patients with DF only indicates that the former patients are genetically predisposed to express higher levels of TNF-alpha. This finding supports studies reporting a possible association between elevated levels of circulating TNF, vascular permeability, and hemorrhage in patients with dengue hemorrhagic fever. PMID- 15361125 TI - Intercellular adhesion molecule-1 polymorphisms in Korean patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is characterized by synovial proliferation and the accumulation of inflammatory cells in the affected joints. Intercellular adhesion molecule-1 (ICAM-1) is readily detected in RA synovial tissues and helps recruit inflammatory cells to the joint. ICAM-1 shows genetic polymorphisms at codons 241 (R241G) and 469 (K469E). In order to investigate the association between ICAM-1 gene polymorphisms and RA, we genotyped ICAM-1 R241G and ICAM-1 K469E polymorphisms in 143 Korean patients with RA, and in 138 healthy controls, by using the polymerase chain reaction-restriction fragment length polymorphism method. No polymorphism of R241G was found in Korean subjects. However, the frequency of the K469 allele was found to be significantly lower in RA patients than in healthy controls. Allele frequency of K469 was lower in RA patient group, compared to that in healthy controls, regardless of the shared epitope status. Distribution of K469E allele frequencies was not different whether the patient had rheumatoid factor, radiographic erosion or extra-articular complications. In conclusion, this study shows lower frequency of the ICAM-1 K469E allele in Korean patients with RA than that in healthy controls. PMID- 15361126 TI - Microgeographic variation of HLA-A, -B, and -DR haplotype frequencies in Tuscany, Italy: implications for recruitment of bone marrow donors. AB - HLA-A/B haplotype frequencies were estimated in a sample of 2355 bone marrow donors born in a subregion of Tuscany (Italy), and the HLA-A, -B, -DR haplotype frequencies were estimated in a subset of 809 individuals. This area was divided in 10 subsamples (two-locus haplotypes), or six subsamples (three-locus haplotypes), all with sample size >50, based on administrative boundaries. A considerable level of heterogeneity of haplotype frequency was present among subsamples; this heterogeneity was associated to a large variation (up to 4-fold) of the number of new donors that must be typed in order to reach 50% chance of finding an HLA-A, -B phenotype of intermediate frequency. Knowledge of the genetic structure of the population at a microgeographic level may be useful in directing the search of specific bone marrow donors. PMID- 15361127 TI - High-resolution typing of HLA-DRB1 locus in the Macedonian population. AB - The Macedonian population is of special interest for HLA anthropological study in the light of unanswered questions regarding its origin and relationship with other populations, especially the neighbouring Balkanians. Two studies have been performed to examine HLA molecular polymorphism in the Macedonian population, so far. The present study is the first to be performed in Macedonia using high resolution sequence-based method for direct HLA typing. The study included 158 unrelated healthy volunteers of Macedonian origin and nationality, having a Christian Orthodox religion. After the simultaneous amplification of exon-2 on both HLA-DRB1 alleles, DNA sequencing was used for genotype assignment. In the 158 samples analysed, all 316 alleles were typed and a total of 29 different DRB1 alleles were detected, with DRB1*1601 being the most frequent allele (14.9%), followed by DRB1*1104 (13.9%). A phylogenetic tree constructed on the basis of the high-resolution data deriving from other populations revealed the clustering of Macedonians together with other Balkan populations (Greeks, Croats, Turks and Romanians) and Sardinians, close to another "European" cluster consisting of the Italian, French, Danish, Polish and Spanish populations. The included African populations grouped on the opposite side of the tree. PMID- 15361128 TI - Cytokine gene polymorphisms in Taiwan. AB - Cytokine gene polymorphisms may affect their transcription, influence their level of production, and may be implicated in inducing susceptibility or resistance to diseases. Cytokine single-nucleotide polymorphisms (SNPs) were used to determine allelic and genotypic frequencies in the Minnan, the Hakka, and in four indigenous tribes: the Ami, the Tsou, the Atayal, and the Tao (or Yami). The following cytokine gene polymorphisms were analyzed: interleukin-1alpha (IL 1alpha) (T/C -889), IL-1beta (C/T -511, T/C +3962), IL-1R (C/T Pst-I 1970), IL 1Ralpha (T/C Mspa1-I 1100), IL-2 (T/G -330, G/T +166), IL-4 (T/G -1098, T/C -590, T/C -33), IL-4Ralpha (G/A +1902), IL-6 (G/C -174, G/A nt565), IL-10 (G/A -1082, C/T -819, C/A -592), IL-12 (C/A -1188), interferon-gamma (A/T UTR 5644), transforming growth factor-beta (C/T codon 10, G/C codon 25), and TNF-alpha (G/A 308, G/A -238). Little differences were observed between the Minnan and the Hakka. On the other hand, the Minnan and Hakka showed significant differences with the indigenous people. PMID- 15361129 TI - Identification of two new HLA alleles: B*3546* and B*5611*. How reliable are the published HLA-B intron 2 sequences? AB - Polymerase chain reaction-sequence-specific primer (PCR-SSP) typing for human leukocyte antigen (HLA)-B in a male 25-year-old Caucasian individual of Iranian origin and in a 42-year-old German Caucasian bone marrow donor revealed reaction patterns that did not agree with any known HLA specificity, thus suggesting in both cases the existence of a novel allele. Sequence-based typing (SBT) after allelic separation revealed the sequences of the new alleles HLA-B*5611 and B*3546. The sequence patterns of both new alleles might have been generated as the results of double crossing over, possibly over several generations. During the analysis of the HLA-B*3546 intron 2 sequence for possible crossing over points, a base insert, an additional G after position 700, was found. This insert was analyzed using SBT and PCR-SSP and was found to be present not only in all samples carrying B*35, but also in all HLA-B specificities tested. It appears that all known HLA-B alleles may contain a G insert at position 700 of intron 2, and that the published intron 2 sequence alignments of the HLA-B locus may contain errors at this position. PMID- 15361130 TI - Identification of new HLA-C alleles by polymerase chain reaction-sequence specific oligonucleotide typing: Cw*0314 and Cw*1511*. AB - In this article, we report two new human leukocyte antigen-C (HLA-C) alleles, HLA Cw*0314 and Cw*1511, which were identified during routine tissue typing of donors for the Australian Bone Marrow Donor Registry and Australian Cord Blood Bank. HLA Cw*0314 shows six codon changes in exon 3 compared to Cw*030401 and shares some sequence homology with Cw*07 alleles. Cw*1511 has two nucleotide changes compared with Cw*150201 in exon 2, both resulting in amino acid changes in the protein sequence. PMID- 15361131 TI - Identification of three novel HLA class I alleles: HLA-B*3928, HLA-B*400104 and HLA-B*4437. AB - Three new human leukocyte antigen (HLA) class I alleles have been identified in the Tissue Typing Laboratory in Sydney, Australia. Sequence analysis of exon 2 and exon 3 of the HLA-B gene revealed the novel polymorphism. A silent substitution of C to T at nucleotide position 369 has been identified for the HLA B*400104 allele when compared to the closest matched allele, HLA-B*400101. The HLA-B*3928 allele was identified with a nucleotide substitution of G to C at position 362 when compared to the closest matched allele, HLA-B*390101, resulting in an amino acid substitution of Arginine to Threonine. A nucleotide substitution of C to G at position 572 resulting in the amino acid change Serine to Tryptophan was identified in the new allele HLA-B*4437, when compared to the closest matched allele HLA-B*440301. Both amino acid substitutions implicate a different specificity and affinity of antigen binding for the alleles HLA-B*3928 and HLA B*4437. PMID- 15361132 TI - Identification of a novel HLA-DPB1 allele--DPB1*0102. AB - In this report, we describe the identification of a novel HLA-DPB1 allele, DPB1*0102, which was first found in a 27-year-old woman and was later detected again in another person, from the Han population of China. According to the nucleotide alignment, DPB1*0102 was the most similar to DPB1*0202, with only one nucleotide substitution from A to G at position 192 among three closely related alleles--DPB1*0202, DPB1*2401 and DPB1*0501. This substitution was a non synonymous one, which caused a change of amino acid from E to K at codon 69 of DP beta-chain in the mature protein. PMID- 15361133 TI - Identification of two novel HLA class II alleles, DQB1*0311 and DQB1*0620. AB - Two novel HLA class II alleles have been identified in routine typing of a kidney transplant patient and a cord blood unit from the Australian Cord Blood Bank in Sydney. Sequence analysis of exon 2 of the DQB1 genes revealed the novel polymorphism. A substitution of A to C at nucleotide position 136 has been identified for the DQB1*0311 allele when compared to the closest-matched allele, DQB1*030201. An identical substitution has also been identified for the DQB1*0620 allele when compared to the closest-matched allele, DQB1*0602. The substitution results in an amino acid change from methionine to leucine at position 46 implicating different specificity and affinity of antigen binding. PMID- 15361134 TI - Identification of a novel HLA-DRB1*12 allele (DRB1*1210) in the Korean population. AB - At least 11 HLA-DRB1*12 alleles have been identified to date. We report a new HLA DRB1*12 allele, DRB1*1210, that was identified in the Korean population. This new allele differs from HLA-DRB1*120101 by a single nucleotide at position 40 (G-->A) in exon 1 that falls within codon--16 (GTT-->ATT). This change results in a single valine to isoleucine amino acid alteration. PMID- 15361135 TI - HLA class-I and HLA class-II phenotypic, gene and haplotypic frequencies in Tunisians by using molecular typing data. AB - The aim of this study is to define a reliable reckoning of gene frequencies and six-locus haplotypic frequencies of HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DQA1, HLA DQB1 and HLA-DPB1 in the Tunisian population. One hundred unrelated random, healthy people originating from various parts of Tunisia were typed for the alleles of the loci mentioned above by using the molecular techniques polymerase chain reaction--hybridization with oligonucleotide probe (PCR-SSO) and sequence specific primers (SSP). The population studied appeared to be in Hardy-Weinberg equilibrium. Allelic frequency distributions were observed at each locus. The most frequent HLA-A alleles were HLA-A*02 (39%) HLA-A*0101 (25%), HLA-A*30 (21%) and HLA-A*2301 (18%). Moreover, HLA-3A*3601, HLA-1A*6601, HLA-1A*3402 and HLA 2A*8001 were found; however, no HLA-A*4301 was detected. For the HLA-B locus, the most common in descending order were HLA-B*44 (22%), HLA-B*5001 (19%), HLA-B*51 (16%) and HLA-B*18 (15%). Among the 28 alleles HLA-Cw detected, HLA-Cw*6 and HLA Cw*7 were highly predominant with the frequencies of 33 and 30%, respectively. For the HLA class-II loci, HLA-DRB1*0701, HLA-DRB1*11, HLA-DRB1*13 and HLA DRB1*03 were the most frequent DR alleles. For the HLA-DPB1, HLA-DPB1*0401, HLA DPB1*0301 and HLA-DPB1*0201 were the most frequent DP alleles. Many haplotypes were in a strong positive-linkage disequilibrium. The most frequent haplotypes for HLA-A, HLA-B, HLA-C and HLA-DRDQ were HLA-A*3301, HLA-B*1402, HLA-Cw*0802, HLA-DRB1*0102, HLA-DQA1*0101 and HLA-DQB1*0501; HLA-A*2402, HLA-B*0801, HLA Cw*0702, HLA-DRB1*0301, HLA-DQA1*0501 and HLA-DQB1*0201; HLA-A*2902, HLA B*4403.1, HLA-Cw*1601, HLA-DRB1*0701, HLA-DQA1*0201 and HLA-DQB1*0202; HLA A*3002, HLA-B*1801, HLA-Cw*0501, HLA-DRB1*0301, HLA-DQA1*0501 and HLA-DQB1*0201, with frequencies between 0.025 and 0.015. These data can be used as control data for HLA disease associations and paternity studies, but they are also important for the evaluation of the probability rate of success in determining the optimal matched donor in unrelated stem transplantation for Tunisian patients or patients of Tunisian origin. PMID- 15361136 TI - Large-scale screening of Arabidopsis circadian clock mutants by a high-throughput real-time bioluminescence monitoring system. AB - Using a high-throughput real-time bioluminescence monitoring system, we screened large numbers of Arabidopsis thaliana mutants for extensively altered circadian rhythms. We constructed reporter genes by fusing a promoter of an Arabidopsis flowering-time gene - either GIGANTEA (GI) or FLOWERING LOCUS T (FT) - to a modified firefly luciferase gene (LUC(+)), and we transferred the fusion gene (P(GI)::LUC(+) or P(FT)::LUC(+)) into the Arabidopsis genome. After mutagenesis with ethyl methanesulfonate, 50 000 M(2) seedlings carrying the P(GI)::LUC(+) and 50 000 carrying P(FT)::LUC(+) were screened their bioluminescence rhythms. We isolated six arrhythmic (AR) mutants and 29 other mutants that showed more than 3 h difference in the period length or phase of rhythms compared with the wild-type strains. The shortest period length was 16 h, the longest 27 h. Five of the six AR mutants carrying P(GI)::LUC(+) showed arrhythmia in bioluminescence rhythms in both constant light and constant dark. These five AR mutants also showed arrhythmia in leaf movement rhythms in constant light. Genetic analysis revealed that each of the five AR mutants carried a recessive mutation in a nuclear gene and the mutations belonged to three complementation groups, and at least one of which was mapped on a novel locus. Our results suggest that the three loci identified here may contain central clock or clock-related genes, at least one of which may be a novel. PMID- 15361137 TI - Isolated plant nuclei as mechanical and thermal sensors involved in calcium signalling. AB - Calcium signals in the nucleus elicit downstream effects that are distinct from those of cytosolic calcium signals. In the present work, we have evaluated the ability of plant nuclei to sense stimuli directly and to convert them into calcium changes. We show that individual mechanical stimulation of isolated nuclei elicits a single calcium transient at acidic pHs, whereas a series of stimulations leads to oscillations whose frequency reflects that of the stimuli. Conversely, at alkaline pHs, nuclei respond to temperature but not to stretch. The stretch- and the temperature-activated processes differ by their sensitivity to pharmacological drugs known to affect ion channel activities in animal cells. Our data demonstrate that isolated nuclei are able to gauge physical parameters of their environment. This might have a profound influence on the functioning of calcium-dependent processes known to control a large array of molecular events in the nucleus. PMID- 15361138 TI - Comprehensive identification of Arabidopsis thaliana MYB transcription factors interacting with R/B-like BHLH proteins. AB - In-depth analysis of protein-protein interaction specificities of the MYB protein family of Arabidopsis thaliana revealed a conserved amino acid signature ([DE]Lx(2)[RK]x(3)Lx(6)Lx(3)R) as the structural basis for interaction between MYB and R/B-like BHLH proteins. The motif has successfully been used to predict new MYB/BHLH interactions for A. thaliana proteins, it allows to discriminate between even closely related MYB proteins and it is conserved amongst higher plants. In A. thaliana, the motif is shared by fourteen R2R3 MYB proteins and six 1R MYB proteins. It is located on helices 1 and 2 of the R3 repeat and forms a characteristic surface-exposed pattern of hydrophobic and charged residues. Single-site mutation of any amino acid of the signature impairs the interaction. Two particular amino acids have been determined to account for most of the interaction stability. Functional specificity of MYB/BHLH complexes was investigated in vivo by a transient DFR promoter activation assay. Residues stabilizing the MYB/BHLH interaction were shown to be critical for promoter activation. By virtue of proved and predicted interaction specificities, this study provides a comprehensive survey of the MYB proteins that interact with R/B like BHLH proteins potentially involved in the TTG1-dependent regulatory interaction network. The results are discussed with respect to multi functionality, specificity and redundancy of MYB and BHLH protein function. PMID- 15361139 TI - Silencing of hydroperoxide lyase and allene oxide synthase reveals substrate and defense signaling crosstalk in Nicotiana attenuata. AB - The fatty acid hydroperoxide (HP) substrates required for the biosynthesis of jasmonic acid (JA) and green leaf volatiles (GLVs) are supplied by separate lipoxygenases (LOX). We silenced the expression of two genes downstream of the LOX: allene oxide synthase (AOS) and HP lyase (HPL) by antisense expression of endogenous genes (NaAOS, NaHPL) in Nicotiana attenuata, in which the biosynthesis of JA is amplified by herbivore-specific elicitors. We report that these elicitors also amplify wound-induced GLV releases, but suppress the wound-induced increase of NaHPL transcripts, suggesting that substrate flux controls GLV biosynthesis. As expected, silencing of NaHPL and NaAOS reduced GLV release and JA accumulation, respectively. Surprisingly, HPL- and AOS-silenced plants had enhanced JA and GLV responses, suggesting substrate 'crosstalk' between these two oxylipin cascades. Plants with depleted GLVs (as-hpl) were less attractive than wild type (WT) or empty vector control plants in choice-tests with native lepidopteran herbivores. In feeding trials, Manduca sexta larvae developed slower on as-hpl plants. The reduced larval consumption and performance, which was not caused by increases in defense responses in as-hpl plants, could be restored to WT levels by the addition of synthetic GLVs, demonstrating that GLVs function as feeding stimulants. Gene expression profiling by cDNA microarray analysis and characterization of several induced defenses in herbivore-elicited as-hpl and as aos plants revealed differential involvement of JA and GLVs in defense signaling. Elicitation of volatile terpenoids (an indirect defense) requires JA signaling, where as trypsin protease inhibitor elicitation (a direct defense) requires both functional JA and GLV cascades. PMID- 15361140 TI - Comprehensive EST analysis of tomato and comparative genomics of fruit ripening. AB - A large tomato expressed sequence tag (EST) dataset (152 635 total) was analyzed to gain insights into differential gene expression among diverse plant tissues representing a range of developmental programs and biological responses. These ESTs were clustered and assembled to a total of 31 012 unique gene sequences. To better understand tomato gene expression at a plant system level and to identify differentially expressed and tissue-specific genes, we developed and implemented a digital expression analysis protocol. By clustering genes according to their relative abundance in the various EST libraries, expression patterns of genes across various tissues were generated and genes with similar patterns were grouped. In addition, tissues themselves were clustered for relatedness based on relative gene expression as a means of validating the integrity of the EST data as representative of relative gene expression. Arabidopsis and grape EST collections were also characterized to facilitate cross-species comparisons where possible. Tomato fruit digital expression data was specifically compared with publicly available grape EST data to gain insight into molecular manifestation of ripening processes across diverse taxa and resulted in identification of common transcription factors not previously associated with ripening. PMID- 15361141 TI - Selective expression of a novel high-affinity transport system for acidic and neutral amino acids in the tapetum cells of Arabidopsis flowers. AB - Within the flower, microsporogenesis represents a major sink for nitrogen, but knowledge on how the imported nitrogen is transferred from the anther cell layers to developing pollen is lacking. Here, we provide information on characterization of a transporter (AtLHT2) that might play an important role in partitioning of amino acids for microspore development. Biochemical analysis in yeast showed that AtLHT2 transports proline and aspartate with high affinity. However, other neutral and acidic amino acids act as strong competitors for proline and aspartate uptake indicating that AtLHT2 generally transports uncharged and negatively charged amino acids. Comparison of the apparent K(m) values of AtLHT2 with previously characterized amino acid transporters clearly demonstrated that AtLHT2 represents a novel high-affinity system for neutral and acidic amino acids. Northern blot analysis showed strong expression of the amino acid transporter in flower buds. Cellular expression could be resolved by using RNA in situ hybridization and in situ RT-PCR methods, which localized AtLHT2 specifically to the tapetum tissue of the anthers. Developing pollen grains are symplasmically isolated from the sporophytic tissue and rely on the nutrients and other compounds secreted from the tapetum cells. Thus, the functional characterization of AtLHT2, together with our expression and localization studies, strongly suggest that in Arabidopsis flowers, AtLHT2 has a critical function in import of neutral and acidic amino acids into the tapetum cells for synthesis of compounds important for microspore structure and in transfer of organic nitrogen to the locule for pollen development. PMID- 15361142 TI - ABF2, an ABRE-binding bZIP factor, is an essential component of glucose signaling and its overexpression affects multiple stress tolerance. AB - Phytohormone abscisic acid (ABA) regulates stress-responsive gene expression during vegetative growth, which is mediated largely by cis-elements sharing the ACGTGGC consensus. Although many transcription factors are known to bind the elements in vitro, only a few have been demonstrated to have in vivo functions and their specific roles in ABA/stress responses are mostly unknown. Here, we report that ABF2, an ABF subfamily member of bZIP proteins interacting with the ABA-responsive elements, is involved in ABA/stress responses. Its overexpression altered ABA sensitivity, dehydration tolerance, and the expression levels of ABA/stress-regulated genes. Furthermore, ABF2 overexpression promoted glucose induced inhibition of seedling development, whereas its mutation impaired glucose response. The reduced sugar sensitivity was not observed with mutants of two other ABF family members, ABF3 and ABF4. Instead, these mutants displayed defects in ABA, salt, and dehydration responses, which were not observed with the abf2 mutant. Our data indicate distinct roles of ABF family members: whereas ABF3 and ABF4 play essential roles in ABA/stress responses, ABF2 is required for normal glucose response. We also show that ABF2 overexpression affects multiple stress tolerance. PMID- 15361143 TI - Citrus fruit bitter flavors: isolation and functional characterization of the gene Cm1,2RhaT encoding a 1,2 rhamnosyltransferase, a key enzyme in the biosynthesis of the bitter flavonoids of citrus. AB - Species of the genus Citrus accumulate large quantities of flavanones that affect fruit flavor and have been documented to benefit human health. Bitter species, such as grapefruit and pummelo, accumulate bitter flavanone-7-O-neohesperidosides responsible, in part, for their characteristic taste. Non-bitter species, such as mandarin and orange, accumulate only tasteless flavanone-7-O-rutinosides. The key flavor-determining step of citrus flavanone-glycoside biosynthesis is catalyzed by rhamnosyltransferases; 1,2 rhamnosyltransferases (1,2RhaT) catalyze biosynthesis of the bitter neohesperidosides, while 1,6 rhamnosyltransferases (1,6RhaT) catalyze biosynthesis of the tasteless rutinosides. We report on the isolation and functional characterization of the gene Cm1,2RhaT from pummelo which encodes a citrus 1,2RhaT. Functional analysis of Cm1,2RhaT recombinant enzyme was conducted by biotransformation of the substrates using transgenic plant cell culture. Flavanones and flavones, but not flavonols, were biotransformed into 7-O-neohesperidosides by the transgenic BY2 tobacco cells expressing recombinant Cm1,2RhaT. Immunoblot analysis established that 1,2RhaT protein was expressed only in the bitter citrus species and that 1,6RhaT enzyme, whose activity was previously documented in non-bitter species, was not cross reactive. Expression of Cm1,2RhaT at the RNA level was prominent in young fruit and leaves, but low in the corresponding mature tissue, thus correlating well with the developmental pattern of accumulation of flavanone-neohesperidosides previously established. Phylogenetic analysis of the flavonoid glycosyltransferase gene family places Cm1,2RhaT on a separate gene cluster together with the only other functionally characterized flavonoid-glucoside rhamnosyltransferase gene, suggesting a common evolutionary origin for rhamnosyltransferases specializing in glycosylation of the sugar moieties of flavonoid glucosides. PMID- 15361144 TI - Nfu2: a scaffold protein required for [4Fe-4S] and ferredoxin iron-sulphur cluster assembly in Arabidopsis chloroplasts. AB - Nfu proteins are candidates to act as scaffold protein in vivo for iron-sulphur cluster biogenesis. In this work, Nfu2 protein function in the chloroplast was investigated in vivo using T-DNA insertion lines disrupted in AtNfu2 gene. Both alleles characterized presented the same dwarf phenotype due to photosynthetic and metabolic limitations. Nfu2 cDNA expression in nfu2.1 mutant rescued this phenotype. Photosynthesis study of these mutants revealed an altered photosystem I (PSI) activity together with a decrease in PSI amount confirmed by immunodetection experiments, and leading to an over reduction of the plastoquinol pool. Decrease of plastid 4Fe-4S sulphite reductase activity correlates with PSI amount decrease and supports an alteration of 4Fe-4S cluster biogenesis in nfu2 chloroplasts. The decrease of electron flow from the PSI is combined with a decrease in ferredoxin amount in nfu2 mutants. Our results are therefore in favour of a requirement of Nfu2 protein for 4Fe-4S and 2Fe-2S ferredoxin cluster assembly, conferring to this protein an important function for plant growth and photosynthesis as demonstrated by nfu2 mutant phenotype. As glutamate synthase and Rieske Fe-S proteins are not affected in nfu2 mutants, these data indicate that different pathways are involved in Fe-S biogenesis in Arabidopsis chloroplasts. PMID- 15361145 TI - Lesions in the mRNA cap-binding gene ABA HYPERSENSITIVE 1 suppress FRIGIDA mediated delayed flowering in Arabidopsis. AB - Recessive mutations that suppress the late-flowering phenotype conferred by FRIGIDA (FRI) and FLOWERING LOCUS C (FLC) and which also result in serrated leaf morphology were identified in T-DNA and fast-neutron mutant populations. Molecular analysis showed that the mutations are caused by lesions in the gene encoding the large subunit of the nuclear mRNA cap-binding protein, ABH1 (ABA hypersensitive1). The suppression of late flowering is caused by the inability of FRI to increase FLC mRNA levels in the abh1 mutant background. The serrated leaf morphology of abh1 is similar to the serrate (se) mutant and, like abh1, se is also a suppressor of FRI-mediated late flowering although it is a weaker suppressor than abh1. Unlike se, in abh1 the rate of leaf production and the number of juvenile leaves are not altered. The abh1 lesion affects several developmental processes, perhaps because the processing of certain mRNAs in these pathways is more sensitive to loss of cap-binding activity than the majority of cellular mRNAs. PMID- 15361146 TI - AtSUC8 and AtSUC9 encode functional sucrose transporters, but the closely related AtSUC6 and AtSUC7 genes encode aberrant proteins in different Arabidopsis ecotypes. AB - Three members of the Arabidopsis sucrose transporter gene family, AtSUC6-AtSUC8 (At5g43610; At1g66570; At2g14670), share a high degree of sequence homology in their coding regions and even in their introns and in their 5'- and 3'-flanking regions. A fourth sucrose transporter gene, AtSUC9 (At5g06170), which is on the same branch of the AtSUC-phylogenetic tree, shows only slightly less sequence homology. Here we present data demonstrating that two genes from this subgroup, AtSUC6 and AtSUC7, encode aberrant proteins and seem to represent sucrose transporter pseudogenes, whereas AtSUC8 and AtSUC9 encode functional sucrose transporters. These results are based on analyses of splice patterns and polymorphic sites between these genes in different Arabidopsis ecotypes, as well as on functional analyses by cDNA expression in baker's yeast. For one of these genes, AtSUC7 (At1g66570), different, ecotype-specific splice patterns were observed in Wassilewskija (Ws), C24, Columbia wild type (Col-0) and Landsberg erecta (Ler). No incorrect splicing and no sequence polymorphism were detected in the cDNAs of AtSUC8 and AtSUC9, which encode functional sucrose transporters and are expressed in floral tissue. Finally, promoter-reporter gene plants and T-DNA insertion lines were analyzed for AtSUC8 and AtSUC9. PMID- 15361147 TI - Crosstalk between elicitor-induced cell death and cell cycle regulation in tobacco BY-2 cells. AB - The molecular links between cell cycle control and the regulation of programmed cell death are largely unknown in plants. Here we studied the relationship between the cell cycle and elicitor-induced cell death using synchronized tobacco BY-2 cells. Flow cytometry and fluorescence microscopy of nuclear DNA, and RNA gel-blot analyses of cell cycle-related genes revealed that the proteinaceous elicitor cryptogein induced cell cycle arrest at the G1 or G2 phase before the induction of cell death. Furthermore, the patterns of cell death induction and defence-related genes were different in different phases of the cell cycle. Constitutive treatment with cryptogein induced cell cycle arrest and cell death at the G1 or G2 phase. With transient treatment for 2 h, cell cycle arrest and cell death were only induced by treatment with the elicitor during the S or G1 phase. By contrast, the elicitor-induced production of reactive oxygen species was observed during all phases of the cell cycle. These results indicate that although recognition of the elicitor signal is cell cycle-independent, the induction of cell cycle arrest and cell death depends on the phase of the cell cycle. PMID- 15361148 TI - A structured mutant population for forward and reverse genetics in Barley (Hordeum vulgare L.). AB - Two large-scale ethylmethanesulfonate (EMS) mutant populations from barley (Hordeum vulgare L.) cv. Optic have been developed to promote both forward and reverse genetics in this crop. Leaf material and seed from approximately 20 000 M(2) plants were individually harvested, freeze-dried and archived. DNA was isolated from 9216 plants from the 20 and 30 mm EMS treatments and assembled into 1152 eight-plant pools. To facilitate PCR-based mutation scanning an approach has been employed that combines cleavage of heteroduplexes using the Cel nuclease (Cel I), post-cleavage intercalating dye labeling and the subsequent detection of cleaved products on a Transgenomic WAVE-HS. The populations were evaluated by screening for induced mutations in two genes of interest and the induced mutations were validated by sequence analysis. To enhance the screening process, 12-16 M(3) progeny from each of the M(2) plants were assessed for visible phenotypes and the data entered into a web accessible database (http://bioinf.scri.sari.ac.uk/distilling/distilling.html). PMID- 15361149 TI - Simultaneous determination of the main metabolites in rice leaves using capillary electrophoresis mass spectrometry and capillary electrophoresis diode array detection. AB - The study of the metabolomics of primary metabolites using conventional chemical analyses requires a high-throughput method. Chemical derivatizations are a prerequisite for gas-chromatographic separation, and a large sample quantity is needed for liquid-chromatographic separation and nuclear magnetic resonance detection systems. Recently, we have developed a capillary electrophoresis-mass spectrometry (CE-MS) technology that can simultaneously quantify a large number of primary metabolites, using only a small quantity of samples, and without any chemical derivatizations. Parallel use of a capillary electrophoresis-diode array detector (CE-DAD) system further enables almost all water-soluble intracellular metabolites to be analyzed. We demonstrate, with rice leaves, a simple and rapid method of sample preparation for CE analysis; using this method, we have successfully measured the levels of 88 main metabolites involved in glycolysis, the tricarboxylic acid cycle, the pentose phosphate pathway, photorespiration, and amino acid biosynthesis. PMID- 15361150 TI - Affinity purification of the tobacco plastid RNA polymerase and in vitro reconstitution of the holoenzyme. AB - We affinity-purified the tobacco plastid-encoded plastid RNA polymerase (PEP) complex by the alpha subunit containing a C-terminal 12 x histidine tag using heparin and Ni(2+) chromatography. The composition of the complex was determined by mass spectrometry after separating the proteins of the >900 kDa complex in blue native and SDS polyacrylamide gels. The purified PEP contained the core alpha, beta, beta', beta" subunits and five major associated proteins of unknown function, but lacked sigma factors required for promoter recognition. The holoenzyme efficiently recognized a plastid psbA promoter when it was reconstituted from the purified PEP and recombinant plastid sigma factors. Reconstitution of a plastid holoenzyme with individual sigma factors will facilitate identification of sigma factor-specific promoter elements. PMID- 15361151 TI - Guest editorial: eliminating head lice: itching to do the research. PMID- 15361152 TI - Lecturer practitioners in UK nursing and midwifery: what is the evidence? A systematic review of the research literature. AB - BACKGROUND: Lecturer practitioner roles have been widely established in the UK, and are seen as having the ability to overcome the theory-practice gap in nursing, as well as offering other benefits including functioning as a link between education and practice. AIMS AND OBJECTIVES: This article systematically reviews the research literature on UK lecturer practitioner roles in nursing and midwifery, in order to construct a picture of the themes that emerge from their national implementation. CONCLUSIONS: Only eight published research studies meeting the inclusion criteria were identified in journals, and five more included from the 'grey literature', totalling 13 suitable research reports. Of these 13 papers, six involved nurses and midwives. Key themes from the literature are outlined and discussed. There is an overwhelming preference for qualitative methodologies, although there is a strong argument for quantitative work in mixed methods studies. RELEVANCE TO CLINICAL PRACTICE: Lecturer practitioner roles can make an important contribution to nursing and midwifery education, but this is problematic. It is essential that managers clarify the purpose, responsibilities, support and review of lecturer practitioner roles if they are to be successful. PMID- 15361153 TI - Nurse prescribers' experiences of prescribing. AB - BACKGROUND: Nurse prescribing has advanced rapidly over the previous decade and is clearly on the agenda for the future. Previous research considers nurse prescribing from the patient's perspective, the medical professions' stance and the legal and ethical implications. However, there is a paucity of literature that explores the experiences of nurse prescribers' within their current role. These experiences need investigating to ensure nurse prescribing is able to advance in ways that provide benefit to nurses and thus provides the impetus for the study. AIM: To explore and review nurse prescribers' experiences of prescribing. DESIGN METHODS: A purposeful sample of seven nurse prescribers currently prescribing within a West Midlands Community Trust underwent minimally structured interviews in this qualitative study. Transcribed interviews were analyzed using thematic analysis. FINDINGS: Four themes were generated from analysis of the interviews, 'patient centred care', 'benefits of nurse prescribing', 'support and role satisfaction' and finally 'prescribing difficulties'. CONCLUSIONS: Nurse prescribers' perceive prescribing as a predominantly positive experience, frequently asserting the advantages that prescribing saves the patient and nurse time, is more convenient for the patient and increases the nurses' autonomy and role satisfaction. However, negative experiences of restrictions to practice as a result of nurse prescribers' formulary limitations and duplication of documentation were also described. RELEVANCE TO CLINICAL PRACTICE: Nurse prescribing is a rapidly evolving area of practice with the potential to advance nursing roles. This research aims to provide an insight into the experiences of current nurse prescribers that may then be disseminated and applied to future practice. PMID- 15361154 TI - Therapeutic and non-therapeutic interpersonal interactions: the patient's perspective. AB - BACKGROUND: This research arose from a concern that the recovery of patients was being hindered by a lack of understanding and emphasis on psychosocial care during hospitalization. AIMS AND OBJECTIVES: This paper describes some of the findings from a study that was undertaken to explore and describe, from the perspective of hospitalized patients in Western Australia, the perceived therapeutic effect of interpersonal interactions that were experienced during hospitalization. DESIGN: Grounded theory method. METHODS: Interviews were conducted with 40 patients who were, or who recently had been, hospitalized. Seventy-eight hours of participant observation were also performed and during that time patients and nurses were observed and informally interviewed. Relevant documentation such as nursing care plans and patient notes were also reviewed. RESULTS: Emotional comfort was identified as a therapeutic state that patients perceived as enhancing their recovery. Personal control was found to be a central feature of emotional comfort and this accounted for the way in which patients interpreted therapeutic and non-therapeutic interpersonal interactions that they encountered during hospitalization. This paper will describe the conditions that patients perceived had either promoted or inhibited their emotional comfort. These were identified as the level of security, level of knowing and level of personal value. These conditions had been affected by the interpersonal interactions that had been experienced. The characteristics of therapeutic and non-therapeutic interpersonal interactions will be described. CONCLUSIONS: The results from this study provide further insight into and understanding of the therapeutic effect of psychosocial aspects of patient care. Specifically, these results emphasize the importance that patients in hospital place on all the interpersonal interactions that they experience during hospitalization. RELEVANCE TO CLINICAL PRACTICE. The identification of the characteristics of interpersonal interactions that facilitated emotional comfort provides directions for enhancing the therapeutic potential of all interpersonal interactions experienced by patients in hospital. PMID- 15361155 TI - Stroke care in the home: the impact of social support on the general health of family caregivers. AB - BACKGROUND: Throughout the stroke rehabilitation process, community care is one of the most important elements. Learning to live with and take care of a family member with a stroke is immensely complex and demanding. Without appropriate support, family caregivers are at risk of their own general health decreasing, thus becoming a patient themselves - the second patient in the family. AIM: The aim of this study was to examine aspects of social support available to family caregivers during the first 12-week transitional period following hospital discharge. DESIGN AND METHOD: This study adopted a cross-sectional descriptive design. Regression analysis was performed for examining the types of social support received and the general health of family caregivers. Open-ended questions were used to gain a better understanding of situation-specific supports including their availability, utilization and eventual satisfaction. RESULTS: Findings confirm that home care for the stroke survivor is heavy and demanding. Health-related care tasks were the most stressful. Around 40% of the family caregivers reported somatic symptoms. Fewer persons in the social network of the family caregivers had provided support compared with the number of support people that caregivers thought available. Two inadequate types of support were tangible support and information support. Professional advice and feedback in relation to home care skills were particularly lacking. CONCLUSION: Findings of this study affirmed that stroke care needs exceed the hospital boundary. Professional support to family caregivers is essential so as to sustain home care and protect the caregiver's health. This is particular important during the first 12 weeks transitional period after hospital discharge. RELEVANCE TO CLINICAL PRACTICE. This study highlighted the implications of social support to nursing practice, especially in predischarge planning and community care. Findings of this study may provide information and data for service planning and community resources co ordination to support community stroke care. PMID- 15361156 TI - Stroke and eating difficulties: long-term experiences. AB - BACKGROUND: Previous studies have shown that eating difficulties after stroke are common and often associated with communication problems. These difficulties, however, have mainly been studied from a professional perspective. Although numerous aspects of dysfunction have been identified, little knowledge exists about the experiences of living with eating difficulties. AIM: To explore how people affected by stroke experience living with eating difficulties, during a prolonged period. DESIGN: Explorative, qualitative case study. METHODS: Repeated interviews and participant observations with three persons 1.5-2 years after their last stroke. Data were analysed using qualitative analysis. RESULTS: Eating difficulties after stroke were experienced as Striving to live a normal life, with the subthemes Abandoned to learn on one's own, Experiences of losses and Feeling dependent. The process of getting back to a life that resembled life before the stroke was experienced as long-lasting and hard work. The informants felt that they were abandoned to manage eating training on their own. The informants experienced a loss of functional eating ability and the ability to perform activities related to food and meals. Feelings of dependence were experienced in mealtime situations. CONCLUSION: Living with eating difficulties after stroke is a complex phenomenon. The informants felt abandoned because of lack of support from the nursing staff. They were left on their own to deal with the difficult process of adjusting to a new way of eating and losses regarding mealtime activities. The combination of repeated interviews and participant observations seemed to be an approach that should be tested in larger studies. RELEVANCE TO CLINICAL PRACTICE: This case study indicates a need for nurses to develop and use evidence-based guidelines for eating training during the continuum of care. Nurses need to assess patient's habits and desires related to eating, and to adjust environment according to patient preferences. PMID- 15361157 TI - Management of fluid and hydration in patients with acute subarachnoid haemorrhage - an action research project. AB - BACKGROUND: The contemporary role of the nurse in managing fluid and hydration in patients is currently ill-defined. Considering the pivotal function nurses have in the delivery of fluid therapies, and the high priority such therapies have in the successful treatment and prevention of secondary brain injury in subarachnoid haemorrhage, the clarification of this role is essential. AIMS AND OBJECTIVES: This research aims to clarify the nurse's role in fluid therapies in relation to subarachnoid haemorrhage. The objectives were to determine how nurses presently see their role in relation to fluid management in patients with subarachnoid haemorrhage, to determine the cues to guide their practice, and how this role corresponds to current patient care. A final objective was to identify how the nurse's role can be maximized to provide optimal patient care. METHODOLOGICAL DESIGN: This project takes an action research approach to examining the nurse's role in the care of patients with subarachnoid haemorrhage. A combination of focus groups, physiological data, nursing and medical documentation and a review of recent literature were used to meet the aims and objectives of the project. RESULTS: The results illustrate that, while nurses involved in the study are knowledgeable about fluid and hydration in subarachnoid haemorrhage, they have an ambiguity surrounding their role. Improvements can be made in the quality of patient care through educational sessions for staff and clarification of medical and nursing interdisciplinary roles. CONCLUSION: This action research project has gone a considerable distance towards begin clarifying this role, and has illustrated clearly that the nurse's role is pivotal to the successful implementation of such treatments. With further education and collaboration with the interdisciplinary team the nurses' role can be expanded to provide optimal, and dynamic patient-centred care. RELEVANCE TO CLINICAL PRACTICE: The results of this study highlight gaps within contemporary nursing and medical approaches to patients with subarachnoid haemorrhage, highlighting areas for improvement. It also begins to clarify the role of the nurse, with evidence of the cues they use to guide their practice. PMID- 15361158 TI - A framework for understanding clinical reasoning in community nursing. AB - BACKGROUND: Clinical reasoning is employed to develop solutions to health needs. The impact of the clinical environment on the organization of knowledge-guiding practice has received limited attention. This gap in the clinical landscape restricts the sharing of decision-making processes. AIMS: Focusing on the community, and specifically the patient's home as a context for nursing, this paper describes the creation of a collective conceptual map for a group of community nurses. There is a twofold aim of exploring the process of exposing and articulating the clinical framework and enhancing and sharing understanding of the clinical paradigm in this context. DESIGN: An interpretive research approach was utilized. Hermeneutic phenomenology guided the level of meaning accessed and constructivism was used to build an educational picture. METHODS: Multiple methods including focus groups, observation and narrative recordings were utilized to collect and analyse research data. RESULTS: All nurses may engage with the same concepts--health, need, care and partnerships--but organized into particular frames by the guiding practice philosophy and service organization. A four-stage framework for understanding clinical reasoning in the community setting is presented. This acknowledges the multi-faceted nature of health, the lived experience of health deficits, and is located in a participation and negotiated model of care. Practice examples are presented to expose the construction of need and response which often occurs in a triadic decision-making process. CONCLUSIONS: Environment of care has significant implications on need identification and response. RELEVANCE TO CLINICAL PRACTICE: Mechanisms to enhance the sharing of clinical reasoning and decision-making transparency are essential to aid inter- and intra-professional communication. Presentation of a clinical reasoning framework exposes the breath of 'signals' encountered in practice and the range of knowledge employed in understanding and responding to patient need. PMID- 15361159 TI - District nursing--sharing an understanding by being present. Experiences of encounters with people with serious chronic illness and their close relatives in their homes. AB - BACKGROUND: In the homecare setting, district nurses assume a heavy responsibility and are involved in a variety of care activities. They view themselves as having a central role in care at home that centres on the development of a relationship with those who are ill and their families. AIM: The aim of this study was to elucidate the meaning of district nurses experiences of encounters with people with serious chronic illness and their close relatives in their homes. A purposive sample of 10 district nurses (female) was interviewed using a narrative approach. METHOD: To achieve the aim, a phenomenological hermeneutic interpretation inspired by the philosophy of Ricoeur was used to interpret the interview text. RESULTS: This study proposes that district nurses' experiences of encounters with people with serious chronic illness and their close relatives in their homes can be understood as district nurses being welcomed into the ill people's privacy, to share their intimacy and their understanding of being ill. This close relationship enables them to alleviate and to console the suffering and loneliness caused by illness. This is expressed in the three themes: being in a close relationship, sharing an understanding and weaving a web of protection. CONCLUSION: It seems that by being entirely present, in a close relationship, district nurses share the experiences of illness and through interpretation of the whole persons' expressions; they share an understanding of this illness experience. In this close relationship, at the home of the ill people and their close relatives, district nurses are available to alleviate people's suffering and loneliness caused by illness. RELEVANCE TO CLINICAL PRACTICE: This study reveals the need to be entirely present in encounters between the district nurses and people with serious chronic illness and their close relatives. This relation makes it possible to establish a shared understanding of the illness experience. Being aware of the importance of this shared understanding within a relationship, will increase the health care personnel's possibility to alleviate and console those suffering of illness. This proposed interpretation could be useful for reflection of care interventions, in education and supervision of district nurses. PMID- 15361160 TI - Families in home care--a resource or a burden? District nurses' beliefs. AB - BACKGROUND: Caring for families in home care is a growing part of Swedish district nurses' professional work. District nurses' facilitative and constraining beliefs about families guide the extent to which families are acknowledged and engaged in the care. AIMS AND OBJECTIVES: The aim of the study was to explore district nurses' beliefs about families in home care. DESIGN: Explorative, descriptive. METHODS: Five district nurses participated in focus group interviews on three separate occasions. Each interview lasted approximately 90 minutes, was audio taped and transcribed verbatim. A thematic content analysis was used for analyses of the data. RESULTS: The result revealed two underpinning beliefs held by the district nurses towards families in home care: families are a resource and Families are a burden. Families could be a resource for the patient, for the other family members as well as for the district nurses themselves. Families could be a resource for the patient both practically and emotionally by e.g. being present and listening. Being open in communication with other family members and district nurses was also considered as a resource. The district nurses considered families as a burden when they were experienced as demanding in various ways, for example, when family members did not act in a way that pleased the district nurses or when family members showed their suffering. CONCLUSIONS: This study highlight some facilitating and constraining beliefs held by district nurses: families can be both a resource and a burden. RELEVANCE TO CLINICAL PRACTICE: It is important that district nurses are aware of what beliefs they hold as their beliefs guide their actions towards the families. PMID- 15361161 TI - Mental health: integrated network and family-oriented model for co-operation between mental health patients, adult mental health services and social services. AB - BACKGROUND: Co-operation between mental health care units and the social services is important in the case of people with social problems who also suffer from mental health problems. However, participation of patients and their families in the treatment process, and co-operation between them and the professionals, are also important. Communication between the professionals, patients and their family members, and the professionals is a crucial factor for co-operation. AIMS AND OBJECTIVES: The aim of this study was to elucidate the experiences and importance of co-operation for the patients. The data consisted of interviews with 22 mental health patients who were also clients of municipal social services. METHOD: The grounded theory approach was used, focusing on the informants' experiences of the integrated network and family-oriented model for co-operation. RESULTS: The findings indicate the importance of the participation of patients and their social networks in psychiatric care or the treatment process. Meetings should be characterized by open and reflexive discussions with all participants' points of view being included, so that fruitful co-operation is possible. However, some negative experiences were also reported, all of which were connected with the professionals' behaviour. CONCLUSIONS: Trust and honesty are essential elements in relations between professionals and psychiatric patients, but it cannot be assumed that they will develop naturally. It is the professionals' responsibility to adjust their behaviour so that these elements can be created in a mutual process between patients and professionals. Multidisciplinary teams are a necessity in family-oriented co-operation between psychiatry and social services, and in a satisfactory caring process. RELEVANCE TO CLINICAL PRACTICE: Nurses' work is often individually oriented and nurses are ruled by routines in their work. The mental health caring process should be seen as a shared process between the patient, his/her human environment and professionals for which nurses need skills to their interaction with patients and their social network. PMID- 15361162 TI - Chinese inpatients' subjective experiences of the helping process as viewed through examination of a nurses' focused, structured therapy group. AB - AIMS AND OBJECTIVES: This study examined Chinese inpatients' views on what aspects of a nurses' focused, structured therapy group worked to help their psychological and interpersonal problems and what traditional Chinese cultural values influenced their viewpoints. METHODS: Nine Chinese inpatients with mental illness participated in the four-session nurses' focused, structured therapy group. After they completed the last session of therapy, they were invited to participate in a structured interview and a semi-structured interview regarding their perceptions of the change mechanisms in nurses' focused, structured group therapy. The semi-structured interviews were recorded and transcribed to be further analysed according to the principal of content analysis. RESULTS: The results indicate that (i) all patients believed that a nurses' focused, structured group psychotherapy enhanced their interpersonal learning and improved the quality of their lives, (ii) traditional Chinese cultural values--those emphasizing the importance of maintaining harmonious interpersonal relationships- influenced the Chinese inpatients' expression of negative emotions in the group and their motivation on interpersonal learning. CONCLUSION: In conclusion, we found that transcultural modification for applying Western group psychotherapy in Chinese culture was needed. The modification included establishing a 'pseudo-kin' or 'own people' relationship among group members and the therapists, organizing warm-up exercises and structured activities, applying projective methods and focusing on the issues of interpersonal relationships and interpersonal problems. RELEVANCE TO CLINICAL PRACTICE: The small sample size of the present study raises questions regarding how representative the views of the sample are with respect to the majority of Chinese inpatients. Nevertheless, this preliminary study revealed a cultural aspect in nursing training that requires significant consideration in order to work effectively with Chinese patients. PMID- 15361163 TI - Concordance with community mental health appointments: service users' reasons for discontinuation. AB - BACKGROUND: Quality issues are being given renewed emphasis through clinical governance and a drive to ensure service users' views underpin health service development. AIMS: To establish service users' reasons for discontinuation of community based mental health appointments in one National Health Service Trust. METHOD: A two-phase survey of all non-completers over a year. Phase one using a structured postal questionnaire. Phase two using structured interviews with respondents to phase one by post, telephone and face to face. RESULTS: A total of 243 discharges because of non-completion were identified by local services over the 12 months of the study and followed up by initial questionnaire. This represents 8.19% of all discharges (2967) within the same period. Forty-four users were engaged and followed up within phase two of the survey. Data were subject to both quantitative and qualitative analysis. CONCLUSIONS: Analysis of responses suggests that the main reasons for non-completion are because of dissatisfaction although the reasons are varied and the interplay between variables is complex. Whilst this user group are not apparently suffering from 'severe mental illness', there is clear, expressed need for a service. RELEVANCE TO CLINICAL PRACTICE: Whoever provides such a service should be responsive to expressed need and a non-medical approach seems to be favoured. If these needs are appropriately met then users are more likely to be engaged and satisfaction is likely to be improved. Although this in itself does not necessarily mean improved clinical outcomes, users are more likely to stay in touch until an agreed discharge. Practical problems of applied health service research are discussed and recommendations are made for a review of referral systems, service delivery and organization with suggestions for further research. PMID- 15361164 TI - Graduate transition scheme for baccalaureate nursing degree students: a pilot project. PMID- 15361165 TI - Commentary on Wilde Larsson B., Larsson G. & Carlson S.R. (2004) Advanced home care: patients' opinions on quality compared with those of family members. PMID- 15361169 TI - An ethnographic study of three mental health triage programs. AB - Triage processes are commonly used to manage the interface between demand for, and supply of, health services. This dimension of service provision is particularly pertinent for mental health services in Australia, where demand outweighs services available. This paper draws on the experiences of using participant observation to explore mental health triage processes. Findings highlight the complexities of the researcher role and benefits of using an ethnographic approach to explore mental health triage patterns. Insider participant observation brings many challenges but also, in this study, enabled the researcher to uncover some roles and processes underpinning triage decisions in mental health services. PMID- 15361170 TI - Borderline pathology in children and adolescents. AB - Mental health nurses have historically been pessimistic about and often unsympathetic towards clients diagnosed with borderline personality disorder. By the time these clients reach adult mental health services their behaviours are often difficult to manage and they often suffer significant re-victimization by health services. Questions need to be raised about how best to avert the consolidation of the problems associated with the disorder. This paper explores the concept of "borderline pathology" in children and adolescents and examines the best available evidence for utilizing an early identification and intervention model for children and adolescents who exhibit this constellation of symptoms. PMID- 15361171 TI - Emotional intelligence in mental health nurses talking about practice. AB - The aim of this study was to explore mental health nurses' experiences of emotional intelligence (EI) in their nursing practice by means of qualitative interviews. The interview questions where developed from the literature on EI. This study used a hermeneutic analysis. Four main themes emerged: relationship with the patient; the substance of supervision; motivation; and responsibility. It was concluded that EI stimulates the search for a deeper understanding of a professional mental health nursing identity. Emotional learning and maturation processes are central to professional competence, that is, personal growth and development. In addition, the moral character of the mental health nurse in relation to clinical practice is of importance. The findings imply multiple types of intelligence related to nursing science as well as further research possibilities within the area of EI. PMID- 15361172 TI - The I-C-A-C framework: uncovering patients' experiences. AB - The use of metaphors to enhance nursing knowledge and intervention has not been thoroughly addressed in a Chinese context. Use of metaphors helps people transform feelings from the inexpressible to the expressible, to revise their view of their own strengths from unaware to aware, and to expand a one dimensional view to a multifaceted one. Based on the therapeutic essence of metaphors, this article proposes the I-C-A-C framework on the use of metaphors, with a case example given to illustrate how nurses can help uncover patients' illness experiences. The I-C-A-C framework has four strategies: I - Inviting the patients to create and share their metaphors; C - Creating many meanings for the metaphors; A - Analysing the metaphors; amd C - Connecting the patients' metaphors to their personal context. Limitations and implications of the framework for nursing practice are suggested. PMID- 15361173 TI - Exploring the impact of the implementation of a nursing clinical development unit program: what outcomes are evident? AB - Nursing Clinical Development Units have become popular as a framework for the facilitation of the professional development of nursing and to contribute to improved outcomes for consumers of health care services. An exploratory evaluation of the Nursing Clinical Development Unit program implemented by the Centre for Psychiatric Nursing Research and Practice was undertaken using a qualitative approach. This paper focuses on the findings relating to outcomes. A number of positive outcomes were identified including: changes within nursing practices; the development of more positive attitudes by nurses, a greater focus on research utilization and evidence-based practice; and an increase in professional development activities, including conference presentations. Despite these positive findings, the need for more structured evaluation and an increased emphasis on dissemination of findings was identified. PMID- 15361174 TI - Mental health survivors: your colleagues. AB - An increasing number of mental health survivors or people who have episodes of mental illness (EOMI) are seeking employment and training as mental health workers (Health Workforce Advisory Committee 2002). Problems for such individuals, as students and in the workforce, will be discussed with special reference to those who are training to complete the New Zealand National Certification in Mental Health Support Work. This article seeks to encourage health professionals to reassess their roles in regard to supported employment for people who have EOMI and to alert them as to the rising number of people who successfully work as trained professionals within mental health areas and have EOMI. The reader is encouraged to reflect on the recovery approach itself and the journey that people who have EOMI encounter when they choose to enter the workforce. This is especially useful to health professionals working in mental health such as consumer groups and nurses. An example of a student who has EOMI is included and the pseudonym "Joe" is applied to this student to protect his identity. PMID- 15361175 TI - Evaluating the impact of a preceptorship course on mental health nursing practice. AB - The crisis in the recruitment of nursing graduates into the mental health nursing field dictates the need for assertive and innovative strategies to promote the positive aspects of this field of nursing. A positive clinical experience has been identified as one of the most important determinants of nursing students' attitudes towards mental health nursing. The Centre for Psychiatric Nursing Research and Practice implemented the course "Preceptorship in Psychiatric Nursing" as a means to increase the skill level within and interest towards the preceptorship role in clinical practice. This paper presents an evaluation of courses conducted between 2000 and 2002. The findings suggest that participants (n = 150) were highly satisfied with the content and delivery of the course, and that it is likely to affect their attitude towards and fulfillment of the preceptorship role. PMID- 15361176 TI - An evaluation of an educational program for clinical supervision. AB - Clinical supervision for mental health nurses has become an area of priority since the implementation of the Enterprise Bargaining Agreement of August 2000 in Victoria. Clinical supervision has been identified as a strategy to improve both the job satisfaction of nurses and the quality of care provided to consumers. A review of the literature suggests that adequate education for supervisors is crucial if this strategy is to prove successful. In response to this identified need, the Centre for Psychiatric Nursing Research and Practice developed the "Clinical Supervision for Health Professionals" program. The evaluation of participants from the first 2 years of the program (n = 63) are mentioned here. The findings suggest a high level of satisfaction with the education provided and that it has been successful in improving participants' attitudes towards, and level of confidence in providing, clinical supervision. PMID- 15361177 TI - Apportioning our time and energy: oral presentation, poster, journal article or other? AB - There is a general expectation for health service employees to present their work in oral or written format to showcase clinical ideas, innovations, service developments, and quality and research initiatives. This research note outlines the types of forums where work can be presented and highlights their relative merits. It is anticipated that this discussion will be of interest to clinicians, managers and researchers when considering where best to present their work. PMID- 15361178 TI - Remaining controversies in the upfront management of advanced ovarian cancer. AB - Ovarian cancer (OC) is one of the leading causes of cancer-related death in women. In the last decades, a lot of energy and resources have been put into a number of clinical trials, with some success. Nevertheless, the prognosis of patients diagnosed with advanced disease remains extremely poor. As research moved forward, some crucial questions with regard to the optimal upfront management of patients with advanced OC (AOC) have remained unanswered. In this article, we review the rationale behind these controversial issues, and provide the levels of evidence supporting the current recommendations for AOC management. PMID- 15361179 TI - Common risk factors of breast and ovarian cancer: recent view. AB - Clinicians, epidemiologists, and public health specialists tend to examine breast and ovarian cancer separately. Although this seems fairly rational and expected, both malignancies are estrogen related and thus share many risk factors. In this review, we investigate the common familial, reproductive, anthropometric, nutritional, and lifestyle risk factors of breast and ovarian cancer. We believe that the parallel examination of the two cancer types could significantly contribute to an improved prevention of "gynecological cancer" as a whole. PMID- 15361180 TI - Histopathologic parameters of prognosis in cervical cancer--a review. AB - Apart from clinical stage and lymph node status, acknowledged to be among the most powerful predictors of outcome in cervical cancer, the determination of prognosis and thereby the need for adjuvant therapy in surgically treated patients currently relies on a variety of histopathologic factors. The role of many of these is controversial. This may be because histopathology is genuinely lacking in sensitivity for predicting tumor behavior in vivo. There is, however, wide variation in histopathologic definitions and criteria. This is probably the major reason for both the lack of reproducibility in the reporting of certain factors and in their diminished value in predicting behavior. Tumor type, grade, vascular invasion, pattern of invasion, and depth are all extremely important prognostic indicators when used individually or as a part of a scoring system. PMID- 15361181 TI - Prophylactic human papillomavirus vaccines: the beginning of the end of cervical cancer. AB - Persistent infection with one of the oncogenic human papillomavirus (HPV) types is a necessity for the development of cervical cancer. By HPV vaccination, cervical cancer could become a very rare disease. Two types of HPV vaccines can be distinguished: (i) therapeutic vaccines which induce cellular immunity targeted against epithelial cells infected with HPV and (ii) prophylactic vaccines inducing virus-neutralizing antibodies protecting against new but not against established infections. At present, several vaccines have been developed and tested in clinical trials. The vaccines are generally well tolerated and highly immunogenic. The current clinical data indicate that prophylactic vaccines are very effective against new persistent infections and the development of cervical intraepithelial lesions. The protection is type specific. However, the follow-up of the vaccination trials is still short. The effect of HPV vaccines on future cancer incidence will only be known after decades of follow-up. This article will address the status of recently terminated phase II and currently running phase III trials with prophylactic HPV vaccines. PMID- 15361182 TI - Assessing economics of treatments for gynecological cancer where clinical effectiveness meets value for money. AB - During the last 10 years, the management of gynecological cancer has been undergoing a great deal of change. This is due to a drive to reduce ineffective treatment and associated morbidity while at the same time maximizing the benefits of currently available treatment. The foundation for this approach has been high quality clinical trials which have been performed in increasing numbers. These trials can provide strong evidence that treatments are equivalent or that a new drug adds superiority to previous treatment. The access that women have to the most effective forms of treatment often depends on the availability of healthcare resources and their affordability within the healthcare system. Healthcare decision makers increasingly require not just clinical effectiveness of treatments but also cost-effectiveness to be demonstrated. While health economic methods have been applied to many forms of cancer treatment and screening, there have been very few rigorous economic studies performed in gynecological cancer. This article discusses how economic analysis can be incorporated into clinical trials and how it can provide the sort of value for money determination that payers of healthcare are now requiring. Economic analysis may add a little to the cost of trials, but in the end, it may increase access to treatment by convincing decision makers of cost-effectiveness. PMID- 15361183 TI - Long-term survival in 463 women treated with platinum analogs for advanced epithelial carcinoma of the ovary: life expectancy compared to women of an age matched normal population. AB - The objective was to assess the long-term survival (5-15 years) in 463 women, with stages IIb-IV epithelial carcinoma of the ovary and to compare their survival with that of a normal population matched for age and sex. Statistical analysis of 463 women, with stages IIb-IV epithelial cancer of the ovary, who were participants in two consecutive North Thames Ovary Group randomized trials, which took place between 1985 and 1994, was performed. The median follow-up period was 10.5 years. The women were treated with debulking surgery, where possible, and adjuvant platinum chemotherapy. One of the randomized groups in the first North Thames trial also received total abdominal radiotherapy. Survival rates at 5, 10, and 15 years were assessed. Prognostic factors for long-term survival were determined using a mathematical model to separate early effects from late effects. The ratio of observed to expected deaths compared to the normal population was calculated. Overall survival at 5 years was 21% (95% confidence intervals 17.5-25%), at 10 years was 13.5% (95% confidence intervals 10.5-17%), and at 15 years was 12% (95% confidence intervals 9-16%). The important prognostic factors for long-term survival were disease-free or minimal residual disease (a single remaining deposit <2 cm) at initial surgery with tumor grade 1 and good performance status. Compared with the normal population (1995 data), the ratio of observed to expected deaths after start of chemotherapy at 5 years was 14.1 (P < 0.001 Fisher's exact test), at 9-10 years 4.9 (P = 0.0033, Fisher's exact test), while in the 11- to 15-year period it had dropped to 2.75 (P = 0.090, Fisher's exact test), which was not significantly different. Patients with advanced cancer of the ovary, who survive 11 years or longer, have a life expectancy which is very similar to that of a normal population of women of the same age. Women with advanced ovarian cancer have an improved chance of long-term survival following treatment if they present with minimal residual disease after primary surgical debulking, grade 1 tumors, and good performance status. PMID- 15361184 TI - Number of residual nodules, better than size, defines optimal surgery in advanced epithelial ovarian cancer. AB - The objective of the present study was to improve the definitions of optimal and suboptimal surgery in ovarian cancer. A retrospective prognostic factor analysis was done in a series of 433 patients with advanced ovarian cancer (stages III and IV) treated during the cisplatine era from 1980 to 1997 with assessment of postoperative residual disease by number and ranges of size of peritoneal nodules. Mean age of patients was 56.7 years. Median follow-up of patients alive was 138 months (range: 6-266 months). Median overall survival was 23 months. Significant prognostic factors for overall survival in univariate analysis were age, the presence of ascites, FIGO stage, treatment period, histological subtype, grade, results of surgery as defined by size and number of residual nodules. In multivariate analysis, quality of surgery defined by no versus few versus numerous residual nodules showed to remain an independent prognostic factor for outcome (P = 0.003), whereas size of residual nodules did not achieve significance. In conclusion, only complete surgery (no residual nodules) confers a real survival advantage. Cytoreduction to few and small nodules is associated with favorable outcome and could be qualified as optimal. Otherwise, cytoreduction leaving numerous nodules, whatever their size, remains suboptimal. Such patients should be considered for neo-adjuvant chemotherapy. PMID- 15361185 TI - Prognostic importance of the time interval from surgery to chemotherapy in treatment of ovarian carcinoma. AB - In a large study on 1,220 patients with ovarian carcinoma in FIGO stages I-IV, the prognostic importance of the time factor for start of postoperative chemotherapy was studied together with other important factors for long-term survival. The patient series was a total geographic material of ovarian carcinoma patients treated during the years 1975-1993. All patients were followed up for 10 years or until death. The 5-year cancer-specific survival rate of the complete series was 50%. Significant and independent prognostic factors with regard to long-term cancer-specific survival were FIGO stage, histology, tumor grade, and completeness of the primary surgery. Special attention was paid to the prognostic importance of the time interval between primary surgery and the first course of chemotherapy. Patient groups with intervals shorter or longer than the median value were compared. In early-stage disease, no significant difference was noted. In advanced and bulky disease, an interval longer than the median value seemed to be beneficial compared with a shorter interval. However, after correction for other prognostic factors, the interval was not a significant factor (P = 0.647) with regard to the cancer-specific survival rate. Therefore, the time factor should not be an important argument for how to best organize the gynecologic oncology service. PMID- 15361186 TI - Whole-abdomen, single-dose consolidation radiotherapy in patients with pathologically confirmed complete remission of advanced ovarian epithelial carcinoma: a long-term survival analysis. AB - The value of consolidation therapy in advanced epithelial ovarian carcinoma patients is controversial. The aim of the present study was to assess the long term survival of patients with a pathologically confirmed complete remission who had consolidation by single-dose, whole-abdominopelvic radiotherapy. Of 96 histologically confirmed stage II-IV epithelial ovarian carcinoma patients who underwent cytoreductive surgery followed by high-dose, platin-based chemotherapy, 57 were in complete clinical remission at the end of therapy and 50 underwent a second-look laparotomy. The study group comprises 32 consecutive patients who had no pathological evidence of disease and who received 800 cGy single-dose, whole abdominal radiotherapy by an 8 MEV linear accelerator in a single fraction. The absolute 5-year survival and the actuarial 10-year survival were 78.7 and 63.3%, respectively. The survival was significantly better in patients who had < or =2 cm residual disease at the completion of the original operation. No severe postradiation complications were encountered. Mild complications were seen in three (9.4%) patients. Our data indicate a favorable long-term survival of patients with a negative second-look laparotomy who had consolidation with single dose, whole-abdominal radiotherapy. These results seem to suggest that a collaborative, prospective, randomized multiarm study is indicated to solve the controversial issue of consolidation therapy. PMID- 15361187 TI - Second-line therapy of advanced ovarian cancer with GnRH analogs. AB - Ovarian cancer is still the first cause of death among female malignancies. The standard treatment adopted in ovarian cancer is a radical surgical treatment or cytoreduction, followed by six courses of platinum-based chemotherapy; second line regimens are associated with severe side effects. GnRH analogs could represent an alternative therapeutical approach. The aim of our study was to evaluate the role of GnRH analogs in the management of platinum-resistant ovarian cancers. We enrolled 12 patients affected by advanced ovarian cancer, previously treated with six courses of platinum-paclitaxel. In second-line therapy, we used leuprolide on 1, 8, and 28 days of treatment. CA 125 levels were recorded for each patient. One case of clinical partial response was obtained (8.3%). Stable disease was diagnosed in three patients (25%). Progression was recorded in eight cases (66.7%). Progression-free survival was 6 months. The treatment was well tolerated by patients. The high tolerability and the results obtained with leuprolide versus platinum in second-line therapy might permit a better use of the analogs for advanced ovarian cancer. PMID- 15361188 TI - Third-line chemotherapy in platinum- and paclitaxel-resistant ovarian, fallopian tube, and primary peritoneal carcinoma patients. AB - Ovarian carcinoma is a malignant disease with a high rate of recurrence, necessitating repeated chemotherapy treatments. We conducted a retrospective study in patients with platinum- and paclitaxel-resistant ovarian, fallopian tubes and primary peritoneal carcinoma patients treated at M.D. Anderson Cancer Center. We evaluated the responses, progression-free intervals, and overall survival duration of 51 patients after third-line chemotherapy treatment. The overall response rate was 16% (eight cases) with 2% complete response rate (one case) and 14% partial response rate (seven cases). Stable disease was achieved in 31% (16 cases). The progression-free intervals of 24 patients who had response and stable disease was 7.4 months (range, 1.4-18.4 months). The median overall survival of all patients was 15.8 months (95% CI, 8.1-23.4 months). The median survival duration of eight responders was not significantly different from that of 43 nonresponders, 18.9 months (95% CI, 2.4-35.4 months) versus 15.8 months (95% CI, 6.4-25.2 months), respectively (P = 0.73). In conclusion, third-line chemotherapy in our study results in a modest response and prolongation of progression-free interval without obvious impact on survival. The decision to utilize third-line chemotherapy will be a balance of the limited efficacy, toxicity of the agents, and the expertise of the clinician. PMID- 15361189 TI - Prognostic significance of microvessel density and vascular endothelial growth factor expression in advanced ovarian serous carcinoma. AB - The aim of the study was to test the prognostic value of the microvessel density (MVD) within the tumor and the vascular endothelial growth factor (VEGF) expression on clinical response to chemotherapy, on brief disease-free interval, and on cause-specific survival in advanced ovarian serous carcinoma. We evaluated 83 ovarian carcinomas homogeneous for stage, type and grade histologic, surgical, and chemotherapeutic treatment. Brief disease-free interval and cause-specific survival rates (Kaplan-Meier method) were compared using the log-rank test. A multivariate analysis (Cox-proportional hazards model) was used to determine the independent effect of each variable on prognosis. Overall 60 and 120 months cause specific survival rates were 27.7% and 2.4%, respectively. The brief disease-free interval rate was 66.2%. In univariate analysis, VEGF (P = 0.0001 and P = 0.016), MVD (P < 0.0005), and the FIGO stage IIIC even more than FIGO stage IIIA (P = 0.01 and P < 0.0005, respectively) were associated with survival and brief disease-free interval, and the residual tumor was associated with survival (P = 0.021). In multivariate analysis, the factors that were independent predictors of survival were MVD (P < 0.0005), VEGF (P = 0.027), and the FIGO stage IIIC even more than FIGO stage IIIA (P = 0.013). Moreover, MVD was an independent predictor also of brief disease relapse (P = 0.001). Both MVD and VEGF were correlated with clinical response to chemotherapy (P = 0.01 and P = 0.037). Our data suggest that MVD and VEGF may have prognostic significance in advanced ovarian serous carcinoma. PMID- 15361190 TI - Antitumor activity of irofulven against human ovarian cancer cell lines, human tumor colony-forming units, and xenografts. AB - The objective of this study was to investigate the cytotoxic activity of irofulven (HMAF, MGI 114), a unique chemotherapeutic agent currently under clinical investigation, in various preclinical models of ovarian cancer. Antiproliferative effects of irofulven in ovarian cancer cell lines and ovarian tumor specimens were characterized in vitro using sulforhodamine B and human tumor colony-forming assays, respectively. Irofulven demonstrated marked activity against a panel of ovarian tumor cell lines, including IGROV1, OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, and SK-OV-3, all of which exhibit various drug resistance mechanisms. In human tumor cloning assays, irofulven inhibited colony formation in surgically derived ovarian tumors at concentrations as low as 0.001 micro g /ml and indicated superior activity in comparison with paclitaxel when tested against the same tumor specimens. The antitumor activity of irofulven compared to that of paclitaxel was also examined using the SK-OV-3 xenograft model. In mice bearing subcutaneously implanted SK-OV-3 tumors, treatment with paclitaxel failed to inhibit tumor growth; whereas mice treated with maximum tolerated doses of irofulven had a 25% partial shrinkage rate, and the remaining animals had a mean tumor growth inhibition of 82%. The potent activity of irofulven against ovarian tumors in vitro and in vivo supports the evaluation of its clinical activity in ovarian cancer. PMID- 15361191 TI - Do the pelvic lymph nodes predict the parametrial status in cervical cancer stages IB-IIA? AB - The aim of this study was to determine whether the pelvic lymph nodes would predict the parametrial status in patients with cervical cancer stages IB1-IIA submitted to radical surgery and pelvic lymphadenectomy. To this end, we evaluated the relationship between positive and negative pelvic lymph nodes and their parametria. Our final purpose was to use this information to recommend the tailoring of the parametrial resection according to the status of pelvic lymph nodes to decrease the morbidity related with radical paratrectomy. From January 1996 to December 2001, 107 consecutive patients with cervical cancer stages IB1 and IIA were primarily treated by radical hysterectomy type III with systematic pelvic lymphadenectomy in a prospective study. Parametria were studied in two sections: the first included the tissue adjacent to the cervix, and the second the distal 2/3. Lymph nodes were routinary processed. Twenty-two patients (20.6%) had positive pelvic nodes and 16 patients (14.9%) had parametrial involvement, mostly by direct extension. Eight patients with positive pelvic nodes (36.4%) had parametrial involvement, whereas among 85 patients with negative pelvic nodes only eight patients (9.4%) had parametrial involvement (P < 0.001), most in internal parametria (62.5%). The sensitivity of pelvic lymph nodes for parametrial involvement was 50% and the positive predictive value was 36.4%, whereas the specificity was 84.6%; and the negative predictive value 90.6%. In the group of negative pelvic lymph nodes, only two patients (2.3%) had parametrial involvement beyond internal parametria. The univariated and multivariated analysis of prognostic factors was always significant but without a significant independent factor for positive parametria. Pelvic lymph nodes appear as good predictors of parametrial status, especially in node-negative patients, and could be used to decrease the paratrectomy in radical surgery. PMID- 15361192 TI - The surgery of 'inguino-femoral' lymph nodes: is it adequate or excessive? AB - In the management of women with any but the earliest vulval carcinoma, the received surgical dogma is that there should be complete removal of all potentially involved lymph nodes in the groin (distal to the inguinal ligament). Traditionally, this has included stripping bare the femoral vessels after unroofing the deep fascia in the proximal thigh (Hunter's canal), although currently a number of surgeons carrying out block dissection of the groin would be less radical. The morbidity of the more extensive block dissection is important, and the need for a procedure of this magnitude has been challenged on the basis that a deep node chain does not extend more distally than the sapheno femoral junction. This study examined possible reasons for this important difference of opinion. In 20 targeted anatomical groin dissections from cadavers, no nodes were identified deep to the deep fascia distal to saphenous opening. However, in the cribriform fascia covering the saphenous opening, some nodes of the superficial group were found within fenestrations of this fascia, which might account for the historic descriptions of deep femoral nodes distal to the sapheno femoral junction. This finding also suggested that an inguinal lymphadenectomy confined to the superficial fascia may fail to include all those nodes normally regarded as being in the superficial inguinal group. Together, these findings also lend support to the contention that neither the removal of deep fascia in the femoral triangle nor its incision, with consequent stripping of the femoral vessels in the thigh, is normally necessary in a radical groin node dissection. PMID- 15361193 TI - Surgical resection of locally recurrent cervical cancer: a single institutional 70 patient series. AB - Pelvic recurrence of cervical cancer is a life-threatening situation and only local control can provide hope for remission. The aim of this study was to evaluate the role of surgery in the treatment of cervical cancer recurrence. This retrospective study analyzed a series of 70 patients who underwent resection of cervix locoregional recurrence. Thirteen patients had palliative salvage surgery for pelvic complications. Twenty-nine resections were considered as curative. Fifty recurrences required pelvic exenterations. The hospital mortality rate was 9% and the morbidity rate was 44%. Overall 5-year actuarial survival rate was 23%. Survival was significantly higher: (a) after curative resection and (b) after centropelvic recurrence resection. Local control was obtained in 48% of the cases and 13 patients are alive with a median follow-up of 75 months. In conclusion, the results of this small and heterogen series seem to justify an attempt to resection for centropelvic recurrences whenever possible. Palliative surgery should be reserved to salvage therapy and highly selected patients. PMID- 15361194 TI - Minilaparotomy for type II and III radical hysterectomy: technique, feasibility, and complications. AB - The objective was to assess the feasibility, the operative and postoperative outcome, and complications in the use of minilaparotomy for type II and III radical hysterectomy (RH) and pelvic lymphadenectomy (PLN) in early-stage cervical/endometrial cancer. A pilot study on 91 consecutive patients submitted to type II and III RH and PLN for early-stage cervical/endometrial cancer was performed between March 2002 and May 2003 in the Division of Gynecologic Oncology (UCSC, Rome). Thirty-two of 91 cases (35.2%) were eligible for minilaparotomy. The mean operative time was 156.7 min, whereas the mean intraoperative estimate of blood loss was 303.7 ml. A mean number of 32.7 pelvic lymph nodes and 6.2 common iliac nodes were removed. Ileus and removal of bladder catheter were on mean postoperative day 2.4 and 3.4, respectively. The mean number of postoperative days spent in the hospital was 3.7. Intra- and postoperative parameters were compared to laparotomy controls and literature data on laparoscopy and Pfannenstiel incision, showing substantially comparable results. Minilaparotomy is acceptable for selected patients undergoing radical abdominal hysterectomy (RAH) and PLN and does not compromise the adequacy of the procedure. It can be considered as an alternative to the classic midline vertical incision or even to the Pfannenstiel incisions and laparoscopy. PMID- 15361195 TI - Prospective phase I/II study of irradiation and concurrent chemotherapy for recurrent cervical cancer after radical hysterectomy. AB - The purpose of the present study was to evaluate the long-term toxicity and efficacy of irradiation and concurrent chemotherapy for patients with a pelvic recurrence of cervical cancer after a hysterectomy. This prospective phase I / II study was designed to administer irradiation and three cycles of concurrent chemotherapy with cisplatin and 5-FU to patients with recurrent cervical cancer confined to the pelvis. Initial therapy was a hysterectomy and none received prior pelvic irradiation. A total of 22 patients were entered into the study. Patients received irradiation and three cycles of concurrent cisplatin and 5-FU. The acute toxicity from chemotherapy and irradiation was grade 3 in 18% and grade 4 in 9%. No patient died from a treatment-related complication. Follow-up times ranged from 7.2 to 17.6 years. At last follow-up, 14 patients died of metastatic cervical cancer and eight were alive. The 10- and 15-year overall survivals were 35%. Long-term complications included leg edema, vesico-vaginal, and recto vaginal fistulae. Pelvic abscesses developed in three of the four patients with a fistula. By logistic regression, the only significant factor for survival was total irradiation dose (P = 0.04). In conclusion, long-term survival with this treatment regimen is possible but is accompanied by significant late toxicity. PMID- 15361196 TI - Conventional 4-field box radiotherapy technique for cancer cervix: potential for geographic miss without CECT scan-based planning. AB - BACKGROUND: The advantage of 4-field radiation to the pelvis is that the use of lateral portals spares a portion of the small bowel anteriorly and rectum posteriorly. The standard lateral portals defined in textbooks are not always adequate especially in advanced cancer cervix. METHODS: An analysis was done to determine adequacy of margins of standard lateral pelvic portals with CECT defined tumor volumes. The study included 40 patients of FIGO stage IIB and IIIB treated definitively for cancer cervix between 1998 and 2000. An inadequate margin was defined if the cervical growth and uterus were not encompassed by the 95% isodose. RESULTS: An inadequate posterior margin was common with bulky disease (P = 0.06) and with retroverted uterus (P = 0.08). Menopausal status, FIGO stage, associated myoma, and age were of no apparent prognostic significance. Bulk retained significant on multivariate analysis. An inadequate anterior margin was common in premenopausal (P = 0.01); anteverted uterus (P = 0.02); associated myoma (P = 0.01); and younger patients (P = 0.03). It was not influenced by bulk or stage. Menopausal status and associated myoma retained significant on multivariate analysis. CONCLUSION: Without the knowledge of precise tumor volume, the 4-field technique with standard portals is potentially risky as it may under dose the tumor through lateral portals and the standard AP/ PA portals are a safer option. PMID- 15361197 TI - Evaluation of tamoxifen in persistent or recurrent nonsquamous cell carcinoma of the cervix: a Gynecologic Oncology Group study. AB - This study was undertaken to estimate the antitumor activity of tamoxifen in patients with persistent or recurrent nonsquamous cell carcinoma of the cervix. Furthermore, the nature and degree of adverse effects from tamoxifen in this cohort of individuals was examined. Tamoxifen citrate was to be administered at a dose of 10 mg per orally twice a day until disease progression or unacceptable side effects prevented further therapy. A total of 34 patients (median age: 49 years) were registered to this trial; two were declared ineligible. Thirty-two patients were evaluable for adverse effects and 27 were evaluable for response. There were only six grades 3 and 4 adverse effects reported: leukopenia (in one patient), anemia (in two), emesis (in one), gastrointestinal distress (in one), and neuropathy (in one). The objective response rate was 11.1%, with one complete and two partial responses. In conclusion, tamoxifen appears to have minimal activity in nonsquamous cell carcinoma of the cervix. PMID- 15361198 TI - Vulvar cancer patients' quality of life: a qualitative assessment. AB - To develop a vulvar cancer-specific quality of life (QOL) subscale to accompany the Functional Assessment of Cancer-General (FACT-G) questionnaire, semistructured interviews were performed with 15 patients treated for vulvar cancer (FIGO stage 0-3). All but one patient, who received chemoradiotherapy, were treated by radical vulvectomy and six patients received a groin lymph node dissection. Patients experienced reductions in several aspects of QOL including emotional functioning, physical functioning, social functioning, sexuality, and body image. Six patients suffered from lymphedema of the legs with a mean severity of 3.5 on a 10-point scale. Four patients reported pruritus (severity rating 8.5). Seven patients expressed a need for more information about the illness and treatment. Only four patients returned to employment after treatment, and all of these patients reported work-related problems. Reductions in sexual functioning were a major concern for five patients, all younger than 65 years. Other topics were groin discomfort after removal of the lymph nodes and disturbance by odor from the vulva. Results of this study revealed vulvar cancer specific reductions in QOL for inclusion in the newly developed vulvar cancer specific subscale. PMID- 15361200 TI - Relation between deranged antioxidant system and cervical neoplasia. AB - The purpose of this study was to compare the plasma levels of the anti-oxidant system and oxidative stress between a cervical neoplasia group and a normal control group and to investigate the relationship between the levels of micronutrients of antioxidant system and various clinicopathological parameters of cervical carcinoma. A cross-sectional sample of 180 women, including 90 normal controls, was recruited from November 2000 to January 2001. Plasma concentration of malondialdehyde (MDA) was spectrophotometrically measured as the parameter of lipid peroxidation. Plasma levels of antioxidant vitamins were analyzed with the help of reverse-phase, high-pressure liquid chromatography, whereas glutathione peroxidase activity was evaluated by using coupled enzyme procedure. While the activity of glutathione peroxidase and the plasma levels of antioxidant vitamins, such as lutein, beta-carotene, lycopene, and zeaxanthin, were all significantly lower, the concentration of MDA was significantly higher in women with cervical intraepithelial neoplasia (CIN) or invasive cervical cancer, compared to those of the normal control group. The status of the plasma antioxidant system showed no significant correlation with the prognostic factors of cervical cancer. These findings suggest a potential role of oxidative stress-induced lipid peroxidation and the impairment of the anti-oxidant system in the pathogenesis of CIN and carcinoma of the uterine cervix. PMID- 15361199 TI - Effect of daidzein on cell growth, cell cycle, and telomerase activity of human cervical cancer in vitro. AB - Phytoestrogens are some plant compounds exhibiting estrogen-like activities. However, some studies have shown that they also affect the growth of some nonhormone-dependent diseases. In this study, daidzein--one of the most common phytoestrogens--was used to investigate its effects on human cervical cancer cells HeLa in vitro. First, the cell growth was measured by using 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Then, the distributions of cell cycle and apoptosis were analyzed with the help of flow cytometry. Finally, the telomerase activity was detected by using real-time quantitative reverse transcription-polymerase chain reaction. The results showed that at the concentrations from 6.25 to 100 micro mol/l, daidzein inhibited the growth of HeLa cells. Flow cytometric analysis showed that cancer cells were arrested at G(0)/G(1) or G(2)/M phase with daidzein. The inductive effects of apoptosis were more obviously observed in low-concentration groups. After HeLa cells were treated with daidzein, the expression of human telomerase catalytic subunit mRNA decreased. These meant that daidzein affected human nonhormone dependent cervical cancer cells in several ways, including cell growth, cell cycle, and telomerase activity in vitro. PMID- 15361201 TI - HPV-DNA, vascular space invasion, and their impact on the clinical outcome in early-stage cervical carcinomas. AB - The present study was designed to analyze the relationship of human papillomavirus (HPV)-DNA, microvessel density, and their impact on clinical outcome in early cervical carcinoma. HPV-DNA was evaluated in 171 cases of cervical carcinoma treated from 1965 to 1990. In 110 cases, the analyses could be performed. A polymerase chain reaction technique was used on paraffin-embedded specimens obtained before the start of therapy. HPV-DNA of any type was detected in 78% (86/110) of all evaluable tumors. HPV16 was the predominant type and was detected in 56% (62/110), HPV18 in 8% (9/110), and HPV35 in 21% (23/110). Patients with tumors containing HPV16 or HPV18 were significantly (P = 0.011) younger than patients with tumors not containing either of these two subtypes. Vascular space invasion and lymph node metastases were observed more frequently in tumors expressing HPV16 and HPV18 (P = 0.002, P = 0.047) than in tumors negative for these HPV strains. Tumors containing HPV16 and HPV18 were significantly (P = 0.012) larger and more frequently (P = 0.005) associated with higher FIGO stages. The cancer-specific survival rate was lower for patients with HPV16- and HPV18-positive tumors, but the difference was not statistically significant. The microvessel density was a non-significant prognostic factor. The overall 5-year survival rate of the complete series was 91%. It was concluded that HPV-DNA was a prognostic factor in early-stage cervical cancer and was associated with the age of the patient, vascular space invasion, lymph node metastases, tumor size, and FIGO stage. PMID- 15361202 TI - Quantification of human papillomavirus DNA in the plasma of patients with cervical cancer. AB - Plasma human papillomavirus (HPV)-DNA level was measured to evaluate the clinical usefulness of circulating DNA for cervical cancer management. DNA extracted from pretreatment plasma of 50 cervical cancer patients and from serial longitudinal plasma of 21 patients was quantified for HPV16/HPV18 by means of quantitative polymerase chain reaction. Another 15 patients with low-grade lesion (LG), 18 patients with high-grade lesion (HG), and 96 normal individuals were studied as controls. Plasma HPV16-DNA was detectable in 50% of cancer patients. The incidence and median level were statistically higher than those in LG patients and normal, but similar to HG patients. Plasma HPV18-DNA was only detected in 6% of cancer patients and 1% of normal. Same type of HPV present in plasma was also detected in its primary tumor; and the level of plasma HPV16-DNA was dependent on the viral load in primary tumor. Plasma HPV-DNA was not detected in 16 of 21 patients after treatment, and those patients had complete response to therapy. HPV-DNA persisted or reappeared in five patients after treatment (one had persistent disease and another had recurrence). Plasma HPV-DNA might be a valuable marker for monitoring therapeutic response and disease progression in cervical cancer. PMID- 15361203 TI - Estrogen receptor, progesterone receptor, and bcl-2 are markers with prognostic significance in CIN III. AB - There are no known biological markers or technologies to predict the natural history of an individual CIN III. The probability of progression is considered greater with the persistence of high-risk human papillomavirus (HPV) infection and age. p53 polymorphism has been associated with cervical carcinogenesis. Hormone-induced cervical cancer is mediated by estrogen receptor (ER) and progesterone receptor (PR). In cervical cancer, increased bcl-2 and Bax immunoreactivity is generally associated with a better prognosis. The purpose of this study was to evaluate the value of HPV 16 and HPV 18 typing and p53 codon polymorphism genotyping by polymerase chain reaction and ER, PR, bcl-2, and Bax expression by immunohistochemistry in predicting the CIN III clinical behavior of CIN III lesions. We studied the expression of these prognostic factors in the CIN III adjacent to squamous cell microinvasive carcinomas of the cervix (MIC) from 29 patients with FIGO stage IA1 cervical cancer and in 25 patients with CIN III and no documented focus of invasion. In the MIC group, only the CIN III was considered at least 2 mm away from the microinvasive complex. The ER, PR, bcl-2, and Bax immunoreactivity was scored as positive (>10% staining cells) and negative (<10% staining cells). No significant difference was observed between MIC and CIN III group concerning HPV infection and p53 polymorphism. The ER, PR, bcl-2, and Bax immunohistochemical expression was stronger and more frequent in the CIN III group. After multivariable analysis, coexpression of ER, PR, and bcl 2 was the only independent factor in defining low risk of progression for CIN III. Our study suggests that coexpression of ER, PR, and bcl-2 may be a useful tool in identifying the CIN III lesions with low risk of progression to cervical cancer. PMID- 15361204 TI - Prospective self-controlled study on prevention of hysteroscopic dissemination in endometrial carcinoma. AB - Patients diagnosed to have endometrial carcinoma without prior hysteroscopic examination were recruited from March 2000 to August 2003. Normal saline was used to distend the uterine cavity during the hysteroscopic examination to look for endocervical spread before the definitive surgical treatment. We performed laparotomy, clamped both fallopian tubes, and collected peritoneal washing before the hysteroscopic examination was performed. Peritoneal washing was collected once more after the hysteroscopic examination. Hysteroscopic assessment was performed in 103 patients. Of them, 10 patients were excluded from the study due to previous history of tubal sterilization or blockage. The final analysis was confined to 93 patients. Positive peritoneal cytology was found in 10 (10.8%) patients and this finding was significantly related to the tumor grading (P = 0.023), adnexal involvement (P = 0.003), cervical invasion (P = 0.01), and the presence of peritoneal seedlings (P = 0.001). In five of the 10 patients with positive peritoneal cytology before the hysteroscopic examination, malignant cells could also be recovered in the peritoneal washing collected after the hysteroscopic examination. For patients with negative peritoneal cytology before hysteroscopy, none exhibited positive peritoneal cytology after the procedure. Our data suggested that complete occlusion of both fallopian tubes can effectively prevent the dissemination of endometrial malignant cells into the peritoneal cavity during hysteroscopy. PMID- 15361205 TI - High-risk endometrial cancer in young indigent women. AB - OBJECTIVE: The aim of this study was to evaluate a different prevalence and clinical pattern of high-risk endometrial cancer in an indigent population of young women. METHODS: Charts of 71 consecutive patients, treated for endometrial adenocarcinoma during a 6-year period, were reviewed. The patients were divided into two groups contingent upon age--(i) those who were below 40 years and (ii) those who were over 40. Based on histological type, grade, and stage, both groups were subdivided into a low, intermediate, or high-risk cancer category. RESULTS: Of the 13 (18.3%) patients in the younger age group, five patients (38.4%) had high-risk endometrial cancer, compared to only eight patients (13.8%) in the older age group. CONCLUSION: In contradiction to previous reports, our results show that a higher proportion of young indigent women diagnosed with endometrial cancer have a high-risk cancer. Delay in diagnosis can explain only some of the discrepancies in the special clinical pattern of endometrial cancer among this population. Other possible explanations include nutritional differences, genetic susceptibility, immunological status, and high-risk behavior. More epidemiological studies are needed for complete understanding of the unfavorable outcome of endometrial cancer in these young women. PMID- 15361206 TI - The clinical value and the cost-effectiveness of follow-up in endometrial cancer patients. AB - The aim of the present article was to evaluate the cost-effectiveness of follow up in endometrial cancer patients. A literature review was performed regarding the studies that addressed routine follow-up of endometrial cancer. For each published study, the costs of the follow-up program were calculated according to Belgium standards. A mean total of 13% relapsed. Symptomatology and clinical examination detected over 83% of the recurrences. The follow-up cost in euro after 5 and 10 years ranged between 127.68 and 2,028.78 and between 207.48 and 2,353.48, respectively. Based on the available data, there is little evidence of routine follow-up improving survival rates. Multiple protocols are used in practice without an evidence base. There is an urgent need for prospective randomized studies to evaluate the value of the current so-called 'standard medical practice of follow-up.' It is to be expected that the cost of follow-up could be reduced considerably, for instance, by tailoring to low- and high-risk groups, or by abandoning routine follow-up. Symptomatic patients, however, should be evaluated immediately. A reduction in the number of visits and examinations would mean an enormous reduction in costs. This economic benefit would be warmly welcomed in the times of increased health costs and decreased budgets. PMID- 15361207 TI - The use of P53, PTEN, and C-erbB-2 to differentiate uterine serous papillary carcinoma from endometrioid endometrial carcinoma. AB - Endometrial adenocarcinoma is the most common pelvic genital malignancy in the western world. The most common subtype of endometrial cancer is endometrioid endometrial carcinoma (EEC), which has a relatively good prognosis. Uterine serous papillary carcinoma (USPC) is also a subtype of endometrial carcinoma. This is an aggressive carcinoma with the majority of patients presenting at stage 3-4 and has a worse prognosis stage for stage when compared with EEC. In addition, the treatment of USPC is more extensive than that of EEC, and therefore definitive diagnosis before surgery ensures the optimum management for the patient. This study aims to determine whether P53, C-erbB-2, and PTEN antibodies have a use in the diagnosis and distinction of these cancers. We created tissue microarrays for 35 cases of EEC and 25 cases of USPC, and then we assessed the immunohistochemical expression of P53, C-erbB-2, and PTEN. There was significantly greater expression of P53 in USPC than that in EEC. However, neither C-erbB-2 nor PTEN showed any significant difference in expression between the two carcinomas. P53 may have a role in the diagnosis of USPC, but neither C erbB-2 nor PTEN would be useful as part of a diagnostic panel. PMID- 15361208 TI - APC, beta-catenin, and E-cadherin and the development of recurrent endometrial carcinoma. AB - Endometrial carcinoma, generally, has a good prognosis. However, in some patients, the tumor appears to behave very aggressively, a course that cannot be explained with histopathological characteristics. More insight into the molecular background can be valuable to clarify these differences in tumor behavior. The three components associated with the Wnt pathway--i.e., adenomatous polyposis coli (APC), beta-catenin, and E-cadherin--were evaluated in a case-control study of 28 patients with stage-I endometrial carcinomas to determine their involvement in the development of recurrent disease. Mutation analysis of the mutation cluster region of the APC gene, determination of gene promoter methylation status of the APC-1A and E-cadherin genes, and immunohistochemical analysis of APC, E cadherin, and beta-catenin were performed using paraffin-embedded tumor tissue. Twenty-one APC gene mutations were detected in 12 of 28 (43%) patients. Only three mutations would result in a stopcodon in the APC gene. APC gene promoter methylation was assessed in 12 of 28 (43%) patients. APC immunostaining was absent in two of 24 (8.3%) patients. The occurrence of APC mutations, APC gene promoter methylation, and APC immunostaining were not predictive for recurrence. No E-cadherin expression was observed in four of 24 patients (17%). E-cadherin gene promoter methylation could not be detected in any of the patients. The absence of E-cadherin expression was predictive for distant metastases, but not for local recurrence. Nuclear localization of beta-catenin was present in nine of 24 (38%) patients and was not predictive for recurrent disease. Involvement of epigenetic and genetic aberrations in APC and beta-catenin genes seems to be of minor importance for the development of local recurrences and distant metastases. Although the number of patients is limited, E-cadherin expression appears to be predictive for the development of distant metastases in endometrial carcinoma. PMID- 15361209 TI - Molecular differences between RER+ and RER- sporadic endometrial carcinomas in a large population-based series. AB - Studies, to date, have suggested that there are distinct molecular differences between microsatellite stable (RER(-)) and unstable (RER(+)) solid tumors, such as colorectal carcinoma. We investigated a range of molecular events including mutation frequency of K-ras, microsatellite instability within the coding region of TGF-beta RII, BAX, and IGF-IIR, loss of expression of p53, hMLH1, hMSH2, hMSH6, and PTEN, and methylation of hMLH1, hMSH2, and PTEN within a large population-based series of sporadic endometrial carcinomas to establish whether there are distinct differences between replication error repair (RER(+)) and RER( ) cases. RER(+) endometrial carcinomas tended to be diploid with normal p53 expression, compared with RER(-) cases. Mutations in TGF-beta RII, IGF-IIR, and BAX were rare, but there was a strong association between mutation and RER(+) status. Methylation and loss of hMLH1 expression were significantly more common in RER(+) cases, as was methylation of PTEN. K-ras mutations were equally frequent in RER(+) and RER(-) cases. Despite the absence of distinct clinicopathological differences between RER(+) and RER(-) cases in this series of sporadic endometrial carcinomas, our results confirm that there are molecular differences between RER(+) and RER(-) cases, but the molecular events occurring in RER(+) endometrial carcinomas differ from those seen in RER(+) colorectal carcinomas. PMID- 15361210 TI - Evaluation and management of brain metastatic patients with high-risk gestational trophoblastic tumors. AB - A retrospective study to evaluate the characteristics of brain metastatic patients with gestational trophoblastic tumors (GTT) and to analyze the results of treatment has been performed. During 1996-2001, 40 patients with metastatic GTT were diagnosed at Vali-e-Asr Hospital, Tehran, Iran. Of them, nine with brain metastases, which were documented with the help of computed tomography scan, were evaluated retrospectively. Eight patients received EMA-EP regimen (etoposide, methotrexate, actinomycin, etoposide, and cisplatinum) and one received EMA-CO (etoposide, methotrexate, actinomycin, cyclophosphamide, and vincristin). All cases received whole brain irradiation therapy concurrently. The median age of the patients at diagnosis was 30 years (range: 17-53). Six of them were of early group (five with symptoms of central nervous system and one was detected during workup) and three were of late group (relapsed group). Five (56%) patients responded to treatment and four (44%) were deceased (three of them belonged to late group). It seems that multi-agent chemotherapy (EMA-EP) concurrently with whole brain irradiation results in acceptable survival rates in GTT patients with brain metastases. PMID- 15361211 TI - Doppler study of myometrium in invasive gestational trophoblastic disease. AB - OBJECTIVE: In this prospective study, we aimed to assess the prognostic and diagnostic role of color Doppler flow of myometrium in patients with invasive gestational trophoblastic disease (GTD). METHODS: Thirty-seven patients, who were enrolled in the study with invasive mole, were assessed with the help of transvaginal color Doppler ultrasound before and after chemotherapy. The place and the size of the myometrial invasions were assessed. RESULTS: Thirty patients of 37 were treated with the help of single-agent chemotherapy--methotrexate (mtx). In this group, the resistance index (RI) ratios ranged between 0.26 and 0.45 and the size of the myometrial invasion varied between 10 and 50 mm. On the other hand, six patients were treated with mtx and actinomycin D combination and one patient was treated with the help of total abdominal hysterectomy. In this group, the RI ratios ranged between 0.16 and 0.25 and the size of the myometrial invasion varied between 60 and 90 mm. Remission was achieved in all patients. CONCLUSION: Transvaginal color Doppler study can easily detect invasive GTD. When the depth and the width of the myometrial invasion increase and when there is a low diastolic/systolic ratio, the number of courses and the need for combination of chemotherapy increase. PMID- 15361212 TI - Is normal beta-hCG regression curve helpful in the diagnosis of persistent trophoblastic disease? AB - OBJECTIVE: The aim of this study was to evaluate the probable usefulness of normal beta-human chorionic gonadotropin (beta-hCG) regression curve in the diagnosis of persistent trophoblastic disease (PTD). METHODS: A log-value regression curve was developed from the means and 95% confidence limits of serial weekly serum beta-hCG titers of 43 patients with uneventful complete hydatidiform moles and 14 patients, who were previously confirmed as PTD. RESULTS: All 14 PTD patients (100%) had abnormal values, beyond normal range, within 4 weeks. beta hCG was in its upper values, compared to normal regression curve at 2.29 +/- 0.19 weeks. This was earlier than plateau or rise detection at 4.21 +/- 0.33 weeks (P < 0.001). Within 3 weeks of evacuation, 13 of 14 (92.86%) PTD patients' beta-hCG values exceeded the normal range, whereas only six of 14 (42%) showed a rise or plateau. CONCLUSION: Our finding indicates that the normal beta-hCG regression curve may be useful for quicker detection of PTD than the plateau or rise of level. PMID- 15361213 TI - The changes of gene expression profiles in hydatidiform mole and choriocarcinoma with hyperplasia of trophoblasts. AB - The purpose of this study is to investigate changes of gene expression profiles in hydatidiform moles (HM) and choriocarcinoma and to explore causes of trophoblastic hyperplasia. Using cDNA microarray, 4,096 genes were analyzed in two pairs of the tissues of HM versus normal villi and in two pairs of normal primary culture trophoblasts versus JAR cell line of choriocarcinoma. The expressions of two genes in normal villi and HM, as well as in JAR and JEG-3, were examined with the help of immunohistochemistry, immunoblot, and reverse transcriptase-polymerase chain reaction in order to confirm the findings of cDNA microarray. Twenty-four genes were upregulated and 65 genes were downregulated in all HM. Four hundred thirty-three genes were upregulated and 380 genes were downregulated in JAR. Forty-six genes were upregulated in both HM and choriocarcinoma, whereas 13 genes were downregulated. Genes associated with the inhibition of cell proliferation were significantly downregulated, whereas genes associated with cell proliferation, malignant transformation, metastasis, and drug resistance were upregulated. Thymidine kinase-1 (TK-1) and small subunit ribonucleotide reductase (RRM-2) were overexpressed in HM, JAR, and JEG-3. The expressions of TK-1 and RRM-2 in moles were positively correlated with proliferative index of trophoblasts. Our results suggest that altered expression of genes exist in HM and choriocarcinoma. Trophoblastic hyperplasia may be involved in the overexpression of DNA synthetic enzymes. PMID- 15361214 TI - Endodermal sinus tumor of the vulva: successfully treated with high-dose chemotherapy. AB - Endodermal sinus tumors (EST) of the lower female genital tract are uncommon malignancies. Most of these tumors are found in the vagina or in the cervix. Only seven cases of EST involving the vulva are reported. We report an eighth case of vulvar EST. The patient recurred and was successfully treated with high-dose chemotherapy and peripheral blood cell transplantation. Because of their location, an early diagnosis of vulvar EST should be possible. Resection of the lesion by using a modified radical vulvectomy followed by cisplatin-based chemotherapy seems the best treatment for these tumors today. Our patient was initially treated with the help of local surgery and adjuvant chemotherapy consisting of three courses of etoposide and cisplatin. Sixteen months after the first diagnosis, pleural and lung metastases were diagnosed. Second-line chemotherapy followed by high-dose chemotherapy with autologous peripheral blood stem cell support was administered after resection of the lung metastases and biopsy of the pleural metastases. There is no evidence of disease 40 months after the diagnosis of the lung and pleural metastases, and 56 months after the primary diagnosis. PMID- 15361215 TI - Invasive adenocarcinoma arising from uterine adenomyosis involving the rectosigmoid colon. AB - We describe a rare case of invasive endometrioid adenocarcinoma arising from uterine adenomyosis involving the rectosigmoid colon. At laparotomy the uterus was densely adherent to the rectosigmoid colon. The final pathologic study of surgical specimens revealed intact endometrium and endometrioid adenocarcinoma scattered diffusely throughout the posterior myometrium with direct invasion into the rectosigmoid colon. There were numerous adenomyotic foci around the carcinoma. This case emphasizes the fact that biopsy findings from a uterus with adenocarcinoma arising from adenomyosis can be false negative. Physicians should keep in mind the possible existence of malignancies arising from adenomyosis when uterine malignancies are clinically suspected but histologic evaluation fails to confirm the diagnosis. PMID- 15361216 TI - Early-stage ovarian carcinoma combined with pulmonary tuberculosis mimicking advanced ovarian cancer: a case report. AB - In patients of ovarian cancer combined with multiple pulmonary nodules, the diagnosis of metastatic ovarian cancer is always considered. However, benign pulmonary conditions can be discovered instead. An 80-year-old female presented with a rapidly growing ovarian mass, elevated serum CA-125, and multiple pulmonary varying-sized nodular lesions. The pretreatment workup of her lung lesions failed to show a malignant cell, and it also failed to show any evidence of tuberculosis or other infectious diseases. After surgery, her disease was allotted to 'stage IV' epithelial ovarian cancer and adjuvant cytotoxic chemotherapy was then used. However, her sputum culture showed positive growth of Mycobacterium tuberculosis 4 weeks later. For fear of reactivation of pulmonary tuberculosis, the anticancer cytotoxic chemotherapy was postponed and the antituberculous treatment was given instead. After 6-month course of antituberculous therapy, no active lung lesion was detectable. In conclusion, infectious or inflammatory conditions can mimic metastatic disease and therefore add to the difficulty of stage determination. We recommend that there must be positive cytologic or pathologic results of lung lesions to allot a case of ovarian cancer to stage IV. Furthermore, whenever pulmonary lesions are seen on imaging, the possibility of diagnoses other than metastatic ovarian cancer should always be considered. PMID- 15361217 TI - Vulvar Paget's disease with underlying adenocarcinoma simulating breast carcinoma: case report and review of the literature. AB - We report a case of extramammary Paget's disease with underlying adenocarcinoma simulating breast carcinoma of the vulva. An 82-year-old woman was found to have a 5 x 3-cm bulky tumor located in the left labium major, infiltrating to the clitoris, left labium minor, and left lateral tissue of the vulva. Small biopsy of the vulva showed intraepidermal proliferation of Paget cells. The patient underwent wide local excision of the vulvar tumor and dissection of left inguinal lymph nodes. Histopathological examination of the resected specimens revealed that Paget cells were distributed singly or tended to form small nests in the epidermis, and that association of these cells with the underlying carcinoma invading to the subcutis could be seen. The underlying carcinoma was composed of squamoid solid nests with central necrotic debris, mimicking 'comedocarcinoma' of the breast. In other areas, the tumor cells were present in tubular formations and solid cords reminiscent of invasive ductal carcinoma of the breast. Immunohistochemically, the Paget cells and the underlying carcinoma cells were positive for carcinoembryonic antigen, epithelial membrane antigen, estrogen receptors, and glandular keratins except for CK 20. We speculate that our case is vulvar Paget's disease presenting as a manifestation of underlying breast carcinoma of the vulva, which might have arisen from either the ectopic breast tissue or anogenital mammary-like glands. PMID- 15361218 TI - Small cell carcinoma of the endometrium: report of three cases and literature review. AB - BACKGROUND: Small cell carcinoma of the endometrium is extremely rare. Aim. We reported three cases of this rare tumor and reviewed the literature. CASES: Case 1 was a 54-year-old woman and case 3 was a 58-year-old woman. Both patients presented with vaginal bleeding. Case 2, a 53-year-old woman, had no symptoms and had a vaginal-cervical smear suspicious for malignancy. All patients underwent surgery and their tumors originated in the endometrium. In all three cases, pathological examination revealed small cell carcinoma of endometrium, and immunohistochemical reactivity for one or more neuroendocrine markers was found in all cases. Under electron microscopy in case 2 and case 3, dense core granules in the cytoplasm of tumor cells were found only in case 3. Case 3 was stage IIIA and died of her disease 12 months after surgery. Both cases 1 and 2 were stage IB and alive with no evidence of disease for 28 months and 9 years, respectively. CONCLUSION: Although the prognosis of small cell carcinoma of endometrium is poor, early detection of this disease may contribute to an improved prognosis. PMID- 15361219 TI - Mullerian adenosarcoma with sarcomatous overgrowth of the cervix presenting as cervical polyp: a case report and review of the literature. AB - An aggressive variant of adenosarcoma, mullerian adenosarcoma with sarcomatous overgrowth (MASO) in the cervix is extremely rare. This variant contains obvious, high-grade sarcoma in addition to a low-grade form. In this report, we describe a case of MASO of the uterine cervix and review the clinical and pathological features of these tumors. The patient was a 37-year-old woman with a cervical polypoid mass, which was morphologically considered as a benign endocervical polyp. Microscopically, polypoid cervical mass showed diffuse and dense malignant spindle cell proliferation around the benign endocervical glands and also an area of markedly anaplastic and pleomorphic spindle cell proliferation, so called, sarcomatous overgrowth. Total abdominal hysterectomy and bilateral salpingo oophorectomy with pelvic lymph node dissection were performed. The patient has been followed-up and neither chemotherapy nor other adjuvant therapies have been administered. At present, she has been clinically free of disease for 9 months since she received surgery. It is extremely rare that MASO of the uterine cervix is presented in premenopausal woman. Gynecologists and pathologists should be aware of the difficulties associated with a delay in the diagnosis of MASO when the tumor is present as a benign looking cervical polyp. PMID- 15361220 TI - Depressed host immunity in a case of metachronous primary uterine papillary serous carcinoma and non-Hodgkin's lymphoma. AB - Metachronous primary malignant tumors of uterine papillary serous carcinoma (UPSC) and non-Hodgkin's lymphoma (NHL) are rare. UPSC is a clinically aggressive and morphologically distinctive variant of endometrial carcinoma. We describe the clinical features of a 63-year-old patient with UPSC that was found 2 years after chemotherapy for malignant lymphoma of neck, stage IV and 5 months after radiation therapy for recurrence. This patient had undergone staging surgery and postoperative radiation for UPSC. One month after completion of radiotherapy, the patient expired due to persistence of the disease. The association between host immunity and UPSC is rarely described in the literature. Immunological profiles of this patient, with compositional changes of natural killer, B, and T cell, dramatically altered the percentage of CD4(+) T cell, CD8(+) T cell, and CD4/CD8 ratio, signifying depressed host immunity. Immunological profile of this patient stressed the issue of depressed host immunity and associated malignancies. PMID- 15361221 TI - Extraovarian nongestational choriocarcinoma in a postmenopausal woman. AB - Primary extrauterine choriocarcinoma especially in the postmenopausal period is very rare. A 69-year-old woman, complaining of back pain, weakness, and severe fatigue with gross hematuria, was found to have paraovarian pelvic mass and underwent laparotomy. At the operation, a mass located in the right adnexal region, extending to the retroperitoneum and within the normal pathway of germ cell migration, was observed. Other peritoneal surfaces, the uterus, both the ovaries, and fallopian tubes were normal. Total abdominal hysterectomy, bilateral salphingo-oophorectomy, and excision of the right adnexal mass were performed. The histopathological report showed an extrauterine, nongonadal pure choriocarcinoma. As single-agent chemotherapy with methotrexate was ineffective, the patient received multiagent chemotherapy and responded well to the treatment. After 18 months following chemotherapy, the patient was disease free. Extraovarian nongestational choriocarcinoma can be seen within the normal pathway of germ cell migration and responds to chemotherapy-like gestational choriocarcinoma. PMID- 15361222 TI - Malignant perivascular epithelioid cell tumor (PEComa) arising in the broad ligament. AB - Malignant perivascular epithelioid cell tumor (PEComa) is an extremely rare mesenchymal neoplasm mostly composed of HMB-45-positive epithelioid cells with clear-to-eosinophilic cytoplasm, a propensity for perivascular distribution and a coexpression of smooth muscle markers. The uterus seems to be one of the most prevalent sites of involvement, although only 14 cases of uterine PEComa have been described. We report the case of a 51-year-old woman with a PEComa arising in the broad ligament. She was treated with total abdominal hysterectomy, bilateral salpingo-oophorectomy, omentectomy, and pelvic radiation, and remains without evidence of disease 15 months after diagnosis. This is, to the best of our knowledge, the first report of a malignant PEComa arising in the broad ligament. To correctly diagnose PEComa, an extensive immunohistochemical panel is essential. As PEComas can behave in an aggressive manner, careful follow-up is warranted. PMID- 15361223 TI - Primary tubal choriocarcinoma. AB - Choriocarcinoma is one of the most serious forms of gestational trophoblastic tumor. It is a malignant tumor from the epithelium of the chorionic villi. The most frequent location site is the uterus. Associated with ectopic pregnancy, it is extremely rare and in general, very aggressive. In 75% of the cases, it items from distant metastasis; therefore, a histological examination of the tubes must be performed in all ectopic pregnancies. Our patient was a 33-year-old woman who was admitted to emergency room (ER) with an intense pain in the right, iliac cavity, and limited genital bleeding. During the exploration, there was abdominal pain, with doubtful signs of peritoneal irritation. The vaginal ultrasound offered an image that was compatible with an extra uterine pregnancy in the left appendages. At emergency, right salpingectomy was performed via laparotomy. The patient was treated with polychemotherapy and contraceptives for a year, with no recurrence of the disease. Control follow-up was performed using beta-human chorionic gonadotropin (HCG) testing on a weekly basis during the first month and then bi-monthly during the first year of follow-up. PMID- 15361224 TI - Comment on "Comparison of serum vascular endothelial growth levels between patients with and without ovarian malignancies.". PMID- 15361226 TI - Asbestos in drinking water does not cause mesothelioma. PMID- 15361227 TI - Surgical pelvic packing as a means of controlling massive intraoperative bleeding during pelvic posterior exenteration--a case report and review of the literature. AB - This is a report of a case of gynecological hemorrhage after a posterior pelvic exenteration in patients with vulvar cancer treated by temporary pelvic packing at the Department of Gynecology of the Medical University in Gdansk. The packing was successful and the sponges were removed after 24 h. Twenty-eight days after the operation, the patient was transferred to the Department of Radiotherapy for supplementary treatment. In patients with severe intraoperative hemorrhage, intra abdominal packing has been successful as a mode of treatment. PMID- 15361228 TI - Immunoparasitology. PMID- 15361229 TI - Role of the Toll/interleukin-1 receptor signaling pathway in host resistance and pathogenesis during infection with protozoan parasites. AB - Different studies have illustrated the activation of the innate immune system during infection with protozoan parasites. Experiments performed in vivo also support the notion that innate immunity has a crucial role in resistance as well as pathogenesis observed during protozoan infections such as malaria, leishmaniasis, toxoplasmosis, and trypanosomiasis. While major advances have been made in the assignment of bacterial molecules as Toll-like receptors (TLRs) agonists as well as defining the role of the Toll/interleukin-1 receptor (TIR) signaling pathway in host resistance to bacterial infection, this research area is now emerging in the field of protozoan parasites. In this review, we discuss the recent studies describing parasite molecules as TLR agonists and those studies indicating the essential role of the TIR-domain bearing molecule named myeloid differentiation factor 88 in host resistance to infection with protozoan parasites. Together, these studies support the hypothesis that the TIR signaling pathway is involved in the initial recognition of protozoan parasites by the immune system of the vertebrate host, early resistance to infection, development of acquired immunity, as well as pathology observed during acute infection with this class of pathogens. PMID- 15361230 TI - Turning it on and off: regulation of dendritic cell function in Toxoplasma gondii infection. AB - Because of its intrinsic virulence, Toxoplasma gondii induces a potent interleukin-12 (IL-12)-dependent cell-mediated immune response that shuts down the growth of the replicative tachyzoite stage, thus promoting host survival and successful transmission through predation. At the same time, this response must be tightly controlled to prevent lethality due to cytokine-mediated immunopathology. Evidence accumulated in recent years suggests that dendritic cells (DCs) play a major role in the initiation of IL-12-driven host resistance and that IL-12 synthesis by DCs is carefully regulated to avoid overproduction. In addition, this work has revealed a critical role for DCs in determining the highly polarized T-helper 1 (Th1)-type response triggered by the parasite. In this review, we summarize our current understanding of how DC function is initiated by Toxoplasma and how parasite-primed DCs drive Th1 effector choice. In addition, we discuss recent findings concerning the pathways responsible for endogenous regulation of DC IL-12 production during T. gondii infection. PMID- 15361231 TI - Dendritic cells, pro-inflammatory responses, and antigen presentation in a rodent malaria infection. AB - An infection of mice with Plasmodium chabaudi is characterized by a rapid and marked inflammatory response with a rapid but regulated production of interleukin 12 (IL-12), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN gamma). Recent studies have shown that dendritic cells (DCs) are activated in vivo in the spleen, are able to process and present malaria antigens during infection, and may provide a source of cytokines that contribute to polarization of the CD4 T-cell response. P. chabaudi-infected erythrocytes are phagocytosed by DCs, and peptides of malaria proteins are presented on major histocompatibility complex (MHC) class II. The complex disulfide-bonded structure of some malaria proteins can impede their processing in DCs, which may affect the magnitude of the CD4 T-cell response and influence T-helper 1 (Th1) or Th2 polarization. DCs exhibit a wide range of responses to parasite-infected erythrocytes depending on their source, their maturational state, and the Plasmodium species or strain. P. chabaudi-infected erythrocytes stimulate an increase in the expression of costimulatory molecules and MHC class II on mouse bone marrow-derived DCs, and they are able to induce the production of pro-inflammatory cytokines such as IL 12, TNF-alpha, and IL-6, thus enhancing the Th1 response of naive T cells. IFN gamma and TNF-alpha play a role in both protective immunity and the pathology of the infection, and the inflammatory disease may be regulated by IL-10 and transforming growth factor-beta. It will therefore be important to elucidate the host and parasite molecules that are involved in activation or suppression of the DCs and to understand the interplay between these opposing forces on the host response in vivo during a malaria infection. PMID- 15361232 TI - New lessons from old pathogens: what parasitic infections have taught us about the role of nuclear factor-kappaB in the regulation of immunity. AB - The nuclear factor-kappaB (NF-kappaB) family of transcription factors is activated by many infectious and inflammatory stimuli. This family regulates the expression of multiple genes, whose products include cytokines, chemokines, adhesion molecules, and antiapoptotic factors that are important components of the innate and adaptive immune response. A prominent role of NF-kappaB transcription factors in resistance to a variety of infectious diseases was revealed by studies with mice that lack individual family members. However, little is known about the basis for these effects or about the role of individual family members during a coordinated immune response to infection. Diverse parasites such as Toxoplasma gondii, Leishmania major, and Trichuris muris provide a unique opportunity to understand the role of the NF-kappaB system in the development of innate and adaptive immunity to these infections. The basis for resistance and susceptibility to these parasites is well understood, and studies using these experimental systems have provided unique insights into the role of NF-kappaB in the regulation of T-helper 1 cell (Th1) and Th2 type responses. It has become clear that NF-kappaB family members have cell lineage specific functions and that their relative importance varies with type of infection as well as route of pathogen entry. Thus, studies with models of parasitic infection have revealed that individual NF-kappaB family members perform distinct, nonoverlapping, and biologically significant functions in the regulation of immunity and inflammation. PMID- 15361233 TI - Requirements for the development of IL-4-producing T cells during intestinal nematode infections: what it takes to make a Th2 cell in vivo. AB - Components of the type 2 immune response may mediate host protection against both helminthic parasites and harmful allergic responses. A central player in this response is the T-helper 2 (Th2) effector cell, which produces interleukin (IL) 4, IL-5, IL-13, and other Th2 cytokines during the primary and memory response. Specific aspects of the parasite that trigger Th2-cell differentiation are not yet defined. Furthermore, the cell types and cell surface and secreted molecules that provide the immune milieu required for the development of Th2 effector cells and also Th2 memory cells are not well understood. They will probably vary with the particular helminth or other antigen inducing the Th2 response. We have used third stage larvae of intestinal nematode parasites as adjuvants to promote naive nonparasite antigen-specific T cells to differentiate into Th2 cells. This model system avoids possible parasite antigen-specific T-cell clones or cross-reactive memory T cells that may preferentially differentiate into Th2 effector cells during the course of infection and confound the stereotypical components of parasite-induced Th2 cell differentiation. We have found that these parasites have a potent adjuvant effect and have used our model system to begin to investigate the events that lead to the development of polarized Th2 cells in vivo. PMID- 15361234 TI - Immune-mediated regulation of chronic intestinal nematode infection. AB - Gastrointestinal nematode infection is extremely prevalent worldwide in humans and animals. Infection levels vary between individuals in infected populations and exhibit a negative binomial distribution, and some individuals appear to be predisposed to certain infection levels. Moreover, infection tends to be chronic, despite evidence for the acquisition of some degree of acquired immunity. The host is subject to constant and repeated antigenic challenge, and individuals vary in the response they make. While a considerable amount of information is emerging on the immunoregulatory mechanisms operating during acute nematode infection from a variety of laboratory model systems, relatively little work has been carried out on the immune mechanisms underlying chronic infection. This review details some of the work that has addressed this important facet of gut nematode infection, highlighting studies from model systems that give insight into the induction of nonprotective immunity, while at the same time avoiding the induction of host-damaging pathology. PMID- 15361235 TI - Helminth parasites--masters of regulation. AB - Immune regulation by parasites is a global concept that includes suppression, diversion, and conversion of the host immune response to the benefit of the pathogen. While many microparasites escape immune attack by antigenic variation or sequestration in specialized niches, helminths appear to thrive in exposed extracellular locations, such as the lymphatics, bloodstream, or gastrointestinal tract. We review here the multiple layers of immunoregulation that have now been discovered in helminth infection and discuss both the cellular and the molecular interactions involved. Key events among the host cell population are dominance of the T-helper 2 cell (Th2) phenotype and the selective loss of effector activity, against a background of regulatory T cells, alternatively activated macrophages, and Th2-inducing dendritic cells. Increasingly, there is evidence of important effects on other innate cell types, particularly mast cells and eosinophils. The sum effect of these changes to host reactivity is to create an anti-inflammatory environment, which is most favorable to parasite survival. We hypothesize therefore that parasites have evolved specific molecular strategies to induce this conducive landscape, and we review the foremost candidate immunomodulators released by helminths, including cytokine homologs, protease inhibitors, and an intriguing set of novel products implicated in immune suppression. PMID- 15361236 TI - Th2 response polarization during infection with the helminth parasite Schistosoma mansoni. AB - T-helper 2 (Th2) cell responses play a critical role in protection against helminth infections. In the case of Schistosoma mansoni, an important helminth parasite of man, data from a mouse model of human disease have shown that Th2 responses are essential to allow host survival. In this infection, parasite eggs are the primary stimulus for Th2 response development. Recent work has shown that egg molecules exert multiple levels of control over the development of host interferon-gamma-associated inflammatory responses. Soluble egg antigen inhibits the ability of dendritic cells to make interleukin-12 and induces Th2-polarized adaptive immune responses that in combination with regulatory T-cell responses effectively limit Th1 response development. In this article, we discuss the factors influencing Th2 response polarization during infection with S. mansoni. PMID- 15361237 TI - Toward an understanding of the interaction between filarial parasites and host antigen-presenting cells. AB - Lymphatic filarial infection, from an immunologic point of view, is one of the most complex parasite infections. Not only are there different clinical manifestations that reflect differing immune responses, but the parasite's multiple stages, each with distinct anatomic tropism, add a compartmental layer of complexity to an already complicated process. Moreover, these parasites have finely tuned immune evasion strategies that enable escape from the innate immune system. As different stages of the parasite interact with different types of antigen-presenting cells that, in turn, may play a significant role in shaping the subsequent adaptive immune response, the focus of this review is to provide insight into the interaction between filarial parasites and antigen-presenting cells with an eye toward understanding how they influence parasite antigen-driven T-cell responses. PMID- 15361238 TI - Interleukin-4- and interleukin-13-mediated host protection against intestinal nematode parasites. AB - Intestinal worm infections characteristically induce T-helper 2 cell (Th2) cytokine production. We reviewed studies performed with mice infected with either of two intestinal nematode parasites, Nippostrongylus brasiliensis or Trichinella spiralis, that evaluate the importance of the Th2 cytokine interleukin-4 (IL-4) and IL-13 in protection against these parasites. These studies demonstrate that while IL-4/IL-13 protect against both parasites by activating signal transducer and activator of transcription 6 (Stat6) through IL-4 receptor alpha (IL-4Ralpha) ligation, Stat6 activation protects against these parasites through different mechanisms. Stat6-dependent gene transcription promotes expulsion of N. brasiliensis solely through effects on non-bone marrow-derived cells that may include enhancement of intestinal smooth muscle contractility, changes in intestinal epithelial cell function, and increased intestinal mucus secretion. In contrast, Stat6 signaling promotes immunity to T. spiralis both through effects on bone marrow-derived cells that can be reproduced by treating mice with IL-4 or IL-13 and through effects on non-bone marrow-derived cells. The former effects appear to include T-cell-dependent induction of intestinal mastocytosis, while the latter sensitize non-bone marrow-derived cells to mast cell-produced mediators. We argue that a limited ability of the host immune system to distinguish among different nematode parasites has led to the evolution of a stereotyped Th2 response that activates a set of effector mechanisms that protects against most intestinal nematode parasites. PMID- 15361239 TI - Immunopathogenesis of schistosomiasis. AB - In schistosomiasis mansoni, the chronic egg-induced granulomatous response in the liver and intestines may eventually cause extensive tissue scarring and development of portal hypertension. Indeed, much of the morbidity and mortality associated with this disease is directly attributable to the deposition of connective tissue elements in affected tissues. Elucidating the mechanisms that regulate the severity of schistosomiasis has been a major research objective over the past several years. Research conducted with DNA microarrays as well as investigations with a variety of gene knock-out mice have been particularly helpful in achieving this goal. A notable accomplishment in the past few years was the identification of interleukin-13 (IL-13) and the IL-13 receptor complex as central regulators of disease progression in schistosomiasis. Liver fibrogenesis is severely decreased in infected IL-13-deficient mice as well as in wildtype animals treated with IL-13 antagonists. In contrast, IL-13 effector function increases dramatically in IL-13 receptor alpha2 (IL-13Ralpha2)-deficient mice. These mice develop severe hepatic fibrosis, fail to downregulate granuloma formation in the chronic phase of S. mansoni infection, and succumb to the disease at an accelerated rate; thus, identifying the 'decoy' IL-13 receptor as a critical life sustaining 'off' switch for tissue damaging egg-induced inflammation. PMID- 15361240 TI - The immunobiology of Th1 polarization in high-pathology schistosomiasis. AB - Schistosomiasis is a serious global helminthic disease, in which the main immunopathology consists of a granulomatous and fibrosing reaction against tissue trapped parasite eggs. The severity of this inflammatory process, the product of a CD4(+) T-cell-mediated immune response against parasite egg antigens, is, however, markedly uneven, both in human patients and among mouse strains in an experimental model. Severe schistosomiasis is associated with persistently elevated pro-inflammatory T-helper-1 (Th1)-type cytokines, whereas milder pathology is present when Th2 cytokines dominate. This scenario is supported by the pronounced pathology resulting from the obliteration of pathways that facilitate Th2 differentiation and by the development of more intense lesions in mouse strains that fail to downregulate the Th1 response. Genetically prone high pathology mice have a higher proportion of CD4(+) T cells in lymph nodes and granulomas, in which the Th1 phenotype is driven by interleukin-12; they also develop a dominant repertoire against peptide 234-246 of the major Sm-p40 egg antigen, utilizing a strikingly restricted T-cell receptor structure that involves Valpha11.3beta8. In turn, low-pathology mice exhibit enhanced CD4(+) T cell apoptosis, which contributes to limit pathology. The definition of distinctive immune profiles associated with polar forms of schistosomiasis opens opportunities for targeted immuno-intervention in individuals suffering from or at risk of severe disease. PMID- 15361241 TI - Interleukin-13 in the skin and interferon-gamma in the liver are key players in immune protection in human schistosomiasis. AB - Immunity against schistosomes includes anti-infection immunity, which is mainly active against invading larvae in the skin, and anti-disease immunity, which controls abnormal fibrosis in tissues invaded by schistosome eggs. Anti-infection immunity is T-helper 2 (Th2) cell-dependent and is controlled by a major genetic locus that is located near the Th2 cytokine locus on chromosome 5q31-q33. Mutations in the gene encoding interleukin (IL)-13 that decrease or increase IL 13 production account, at least in part, for that genetic control. In contrast, protection against hepatic fibrosis is dependent on interferon (IFN)-gamma and is controlled by a major genetic locus that is located on 6q23, near the gene encoding the IFN-gamma receptor beta chain. Mutations that modulate IFN-gamma gene transcription are associated with different susceptibility to disease. These data indicate that IL-13 in the skin and IFN-gamma in the liver are key players in protective immunity against schistosomes. These roles relate to the high anti fibrogenic activities of IFN-gamma and to the unique ability of IL-13 in Th2 priming in the skin and in the mobilization of eosinophils in tissues. The coexistence of strong IFN-gamma and IL-13-mediated immune responses in the same subject may involve the compartmentalization of the anti-schistosome immune response between the skin and the liver. PMID- 15361242 TI - Manipulation of mitogen-activated protein kinase/nuclear factor-kappaB-signaling cascades during intracellular Toxoplasma gondii infection. AB - The intracellular protozoan Toxoplasma gondii exerts profound effects on nuclear factor-kappaB (NF-kappaB)- and mitogen-activated protein kinase (MAPK)-signaling cascades in macrophages. During early infection, nuclear translocation of NF kappaB is blocked, and later, the cells display defects in lipopolysaccharide (LPS)-induced MAPK phosphorylation after undergoing initial activation in response to Toxoplasma itself. Infected macrophages that are subjected to triggering through Toll-like receptor 4 (TLR4) with LPS display defective production of tumor necrosis factor-alpha and IL-12 (IL-12) that likely reflects interference with NF-kappaB- and MAPK-signaling cascades. Nevertheless, T. gondii possesses molecules that themselves induce eventual proinflammatory cytokine synthesis. For interleukin-12, this occurs through both myeloid differentiation factor 88-dependent and chemokine receptor CCR5-dependent pathways. The balance between activation and interference with proinflammatory signaling is likely to reflect the need to achieve an appropriate level of immunity that allows the host and parasite to maintain a stable interaction. PMID- 15361243 TI - Does the Leishmania major paradigm of pathogenesis and protection hold for New World cutaneous leishmaniases or the visceral disease? AB - Parasitic protozoa of the genus Leishmania have provided a useful perspective for immunologists in terms of host defense mechanisms critical for the resolution of infection caused by intracellular pathogens. These organisms, which normally reside in a late endosomal, major histocompatibility complex (MHC) class II(+) compartment within host macrophages cells, require CD4(+) T-cell responses for the control of disease. The paradigm for the CD4(+) T-helper 1 (Th1)/Th2 dichotomy is largely based on the curing/non-curing responses, respectively, to Leishmania major infection. However, this genus of parasitic protozoa is evolutionarily diverse, with the cutaneous disease-causing organisms of the Old World (L. major) and New World (Leishmania mexicana/ Leishmania amazonensis) having diverged 40-80 million years ago. Further adaptations to survive within the visceral organs (for Leishmania donovani, Leishmania chagasi, and Leishmania infantum) must have been required. Consequently, significant differences in host parasite interactions have evolved. Different virulence factors have been identified for distinct Leishmania species, and there are profound differences in the immune mechanisms that mediate susceptibility/resistance to infection and in the pathology associated with disease. These variations not only point to interesting features of the host-pathogen interaction and immunobiology of this genus of parasitic protozoa, but also have important implications for immunotherapy and vaccine development. PMID- 15361244 TI - Re-examination of the immunosuppressive mechanisms mediating non-cure of Leishmania infection in mice. AB - The interleukin (IL)-4 driven, polarized T-helper 2 cell (Th2) response that controls non-healing infection with Leishmania major in BALB/c mice has long been embraced as the underlying principle with which to consider the pathogenesis of non-healing and systemic forms of leishmaniasis in humans. The inability, however, to reveal a Th2 polarity associated with non-curing clinical disease has suggested that alternative cells and cytokines are involved in susceptibility. In this review, various mouse models of non-curing infection with L. major and other Leishmania species are re-examined in the context of the suppression mediated by IL-10 and regulatory T (Treg) cells. These activities are revealed in L. major infected BALB/c IL-4 knockout (KO) and IL-4Ralpha KO mice and especially in non cure resistant mice that do not default to a Th2 pathway as a result of inherent defects in Th1 differentiation. In contrast to the extreme BALB/c susceptibility arising from an aberrant Th2 response, non-cure in resistant mice arises from an imbalance in Treg cells that are activated in the context of an ongoing Th1 response and whose primary function may be to suppress the immunopathology associated with persistent antiparasite responses in infected tissues. PMID- 15361245 TI - The immunopathology of experimental visceral leishmaniasis. AB - Experimental murine infection with the parasites that cause human visceral leishmaniasis (VL) results in the establishment of infection in the liver, spleen, and bone marrow. In most strains of mice, parasites are eventually cleared from the liver, and hepatic resistance to infection results from a coordinated host response involving a broad range of effector and regulatory pathways targeted within defined tissue structures called granulomas. In contrast, parasites persist in the spleen and bone marrow by mechanisms that are less well understood. Parasite persistence is accompanied by the failure of granuloma formation and by a variety of pathologic changes, including splenomegaly, disruption of lymphoid tissue microarchitecture, and enhanced hematopoietic activity. Here, we review the salient features of these distinct tissue responses and highlight the varied roles that cytokines of the tumor necrosis factor family play in immunity to this infection. In addition, we also discuss recent studies aimed at understanding how splenomegaly affects the survival and function of memory cells specific for heterologous antigens, an issue of considerable importance for our understanding of the disease-associated increase in secondary infections characteristic of human VL. PMID- 15361246 TI - The immunological challenge to developing a vaccine to the blood stages of malaria parasites. AB - Twenty-one years after malaria antigens were first cloned, a vaccine still appears to be a long way off. There have been periods of great excitement, and in model systems, subunit vaccine homologs can induce robust protection. However, significant challenges exist concerning antigenic variation and polymorphism, immunological non-responsiveness to individual vaccine antigens, parasite-induced apoptosis of immune effector and memory cells, and immune deviation as a result of maternal immunity and alterations of dendritic cell function. Novel approaches will be required. This review addresses some of the approaches that might present malaria antigens in a way designed to induce superior immune responses or that target novel conserved epitopes. Cell-mediated immunity, acting independently of antibody, may exert potent anti-parasite effects, and identification of multiple target antigens/epitopes could lead to the development of vaccines with profound efficacy. PMID- 15361247 TI - Does malaria suffer from lack of memory? AB - It is widely perceived that immunity to malaria is, to an extent, defective and that one component of this defective immune response is the inability to induce or maintain long-term memory responses. If true, this is likely to pose problems for development of an effective vaccine against malaria. In this article, we critically review and challenge this interpretation of the epidemiological and experimental evidence. While evasion and modulation of host immune responses clearly occurs and naturally acquired immunity is far from optimal, mechanisms to control blood-stage parasites are acquired and maintained by individuals living in endemic areas, allowing parasite density to be kept below the threshold for induction of acute disease. Furthermore, protective immunity to severe pathology is achieved relatively rapidly and is maintained in the absence of boosting by re infection. Nevertheless, there are significant challenges to overcome. The need for multiple infections to acquire immunity means that young children remain at risk of infection for far too long. Persistent or frequent exposure to antigen seems to be required to maintain anti-parasite immunity (premunition). Lastly, pre-erythrocytic and sexual stages of the life cycle are poorly immunogenic, and there is little evidence of effective pre-erythrocytic or transmission-blocking immunity at the population level. While these problems might theoretically be due to defective immunological memory, we suggest alternative explanations. Moreover, we question the extent to which these problems are malaria-specific rather than generic (i.e. result from inherent limitations of the vertebrate immune system). PMID- 15361248 TI - Effector and memory CD8+ T cells as seen in immunity to malaria. AB - Transgenic (Tg) mice carrying a T-cell receptor (TCR) specific for a CD8(+) T cell epitope expressed in pre-erythrocytic stages of Plasmodium yoelii has proven to be a valuable tool to advance our understanding of this anti-parasite T-cell response, as it occurs in vivo. The visualization of CD8(+) T cells in vivo and ex vivo greatly facilitated research aimed at characterizing basic features of this T-cell response such as the kinetics of differentiation and proliferation and the in vivo antigen presentation. Importantly, this research unveiled the existence of early self-regulatory mechanisms controlling the magnitude of the CD8(+) T-cell response and also identified CD4(+) T cells as critical elements in the development of memory populations. This review discusses our recent research using Tg mice and highlights our progress in understanding the CD8(+) T-cell mediated immunity against malaria liver stages. PMID- 15361249 TI - Generation, specificity, and function of CD8+ T cells in Trypanosoma cruzi infection. AB - CD8(+) T cells are crucial to the control of Trypanosoma cruzi infection and probably act via multiple mechanisms, the most important being the production of interferon-gamma (IFN-gamma). In the absence of CD8(+) T cells, mice quickly succumb to the infection or develop a more severe chronic disease. Reduced production of IFN-gamma by CD8(+) T cells is also associated with increased severity of chagasic disease in humans. CD8(+) T cells in chronic T. cruzi infection are maintained as effector memory cells, undergo rapid expansion, and demonstrate effector functions following re-exposure to antigen. However, the initial generation of T. cruzi-specific CD8(+) T-cell responses appears to be relatively slow to develop. In addition, the expression of the effector function of the CD8(+) T cells is compromised in some tissues, particularly in muscle. The targets of effective CD8(+) T-cell responses in T. cruzi infection are multiple and varied, and they represent some of the best vaccine candidates described to date. Further analysis of CD8(+) T cells will provide insight into the disease process in T. cruzi infection and should identify methods to assess and enhance immunity to T. cruzi infection and protection from the symptoms of Chagas' disease. PMID- 15361250 TI - The development of effector and memory T cells in cutaneous leishmaniasis: the implications for vaccine development. AB - Leishmania major infections induce the development of a CD4(+) T-helper 1 (Th1) response that not only controls the primary infection but also results in life long immunity to reinfection. How that immunity is maintained is unknown, although because of the existence of infection-induced immunity, there has been an assumption that the development of a vaccine against leishmaniasis would be relatively easy. This has turned out not to be the case. One problem has been the finding that a large part of the immunity induced by a primary infection depends upon the presence of persistent parasites. Nevertheless, there are ample situations where immunologic memory persists without the continued presence of antigen, providing the prospect that a non-live vaccine for leishmaniasis can be developed. To do so will require an understanding of the events involved in the development of an effective protective T-cell response and, more importantly, an understanding of how to maintain that response. Here, we review work from our laboratory, describing how Th1 cells develop in L. major-infected mice, the nature of the memory T cells that provide protection to reinfection, and how that information may be utilized in the development of vaccines. PMID- 15361251 TI - Potential cellular conformations of the CCN3(NOV) protein. AB - AIM: To study the cellular distribution of CCN3(NOV) and to determine if the carboxyterminus of CCN3 is hidden or masked due to high affinity interactions with other partners. CCN3 was detected using affinity purified antibodies (anti K19M-AF) as well as a Protein A purified anti-K19M antibodies (anti-K19M IgG) against a C-terminal 19-aminoacid peptide (K19M) of human CCN3 protein. The antibodies were applied in indirect immunofluorescence tests and immunoenzyme assays on glial tumor cell line, G59, and its CCN3-transfected variant G59/540 and the adrenocortical cell line, NCI-H295R. RESULTS: Anti-K19M-AF antibodies reacted against K19M peptide in ELISA and recognized two bands of 51 kDa and 30 kDa in H295R (adrenocortical carcinoma) cell culture supernatants by immunoblotting. H295R culture supernatants which contained CCN3 as shown by immunoblotting did not react with anti-CCN3 antibodies in liquid phase. Anti-CCN3 antibodies stained the surface membranes of non-permeabilized H295R and cytoplasm in permeabilized H295R cells. Similarly, anti-CCN3 stained surface membranes of G59/540, but did not react with G59 cells. Prominent cytoplasmic staining was observed in G59/540, as well as the cell footprints of G59/540 and H295R were strongly labeled. CONCLUSIONS: The K19M-AF antibody directed against the C terminal 19-aminoacid peptide of CCN3 recognized the secreted protein under denaturing conditions. However, the C-terminal motif of secreted CCN3 was not accessible to K19M-AF in liquid phase. These anti-CCN3 antibodies stained CCN3 protein which was localized to cytoplasmic stores, cell membranes and extracellular matrix. This would suggest that cytoplasmic and cell membrane bound CCN3 has an exposed C-terminus while secreted CCN3 has a sequestered C-terminus which could be due to interaction with other proteins or itself (dimerization). Thus the K19M-AF antibodies revealed at least two conformational states of the native CCN3 protein. PMID- 15361252 TI - Health-related quality of life as a predictor of pediatric healthcare costs: a two-year prospective cohort analysis. AB - BACKGROUND: The objective of this study was to test the primary hypothesis that parent proxy-report of pediatric health-related quality of life (HRQL) would prospectively predict pediatric healthcare costs over a two-year period. The exploratory hypothesis tested anticipated that a relatively small group of children would account for a disproportionately large percent of healthcare costs. METHODS: 317 children (157 girls) ages 2 to 18 years, members of a managed care health plan with prospective payment participated in a two-year prospective longitudinal study. At Time 1, parents reported child HRQL using the Pediatric Quality of Life Inventory (PedsQL 4.0) Generic Core Scales, and chronic health condition status. Costs, based on health plan utilization claims and encounters, were derived for 6, 12, and 24 months. RESULTS: In multiple linear regression equations, Time 1 parent proxy-reported HRQL prospectively accounted for significant variance in healthcare costs at 6, 12, and 24 months. Adjusted regression models that included both HRQL scores and chronic health condition status accounted for 10.1%, 14.4%, and 21.2% of the variance in healthcare costs at 6, 12, and 24 months. Parent proxy-reported HRQL and chronic health condition status together defined a 'high risk' group, constituting 8.7% of the sample and accounting for 37.4%, 59.2%, and 62% of healthcare costs at 6, 12, and 24 months. The high risk group's per member per month healthcare costs were, on average, 12 times that of other enrollees' at 24 months. CONCLUSIONS: While these findings should be further tested in a larger sample, our data suggest that parent proxy reported HRQL can be used to prospectively predict healthcare costs. When combined with chronic health condition status, parent proxy-reported HRQL can identify an at risk group of children as candidates for proactive care coordination. PMID- 15361253 TI - Fusion of green fluorescent protein to the C-terminus of granulysin alters its intracellular localization in comparison to the native molecule. AB - The engineering of green fluorescent protein (GFP) fusion constructs in order to visibly tag a protein of interest has become a commonly used cell biology technique. Although caveats to this approach are obvious, literature reports in which the chimeric molecule behaves differently than the native molecule are scant. This brief report describes one such case. Granulysin, a small lytic and antimicrobial protein produced by cytotoxic lymphocytes, traffics to the regulated secretory system and is subsequently released from cells upon proper stimulus. In an attempt to elucidate mechanisms by which it accumulates in and is released from cytolytic granules, GFP was fused to the C-terminus of granulysin and expressed in an NK cell line. A control construct expressing the native protein was similarly expressed. The data demonstrate that, while the fusion protein is expressed and secreted, its subcellular localization is altered in comparison to native granulysin. Thus, the addition of GFP to the C-terminus of granulysin obscures the signal(s) that cytotoxic lymphocytes use to sort it to the regulated secretory pathway despite its normal biosynthesis and secretion. This example is offered as a cautionary account for other researchers who contemplate using this technology. PMID- 15361254 TI - Analysis of cardiac signals using spatial filling index and time-frequency domain. AB - BACKGROUND: Analysis of heart rate variation (HRV) has become a popular noninvasive tool for assessing the activities of the autonomic nervous system (ANS). HRV analysis is based on the concept that fast fluctuations may specifically reflect changes of sympathetic and vagal activity. It shows that the structure generating the signal is not simply linear, but also involves nonlinear contributions. These signals are essentially non-stationary; may contain indicators of current disease, or even warnings about impending diseases. The indicators may be present at all times or may occur at random in the time scale. However, to study and pinpoint abnormalities in voluminous data collected over several hours is strenuous and time consuming. METHODS: This paper presents the spatial filling index and time-frequency analysis of heart rate variability signal for disease identification. Renyi's entropy is evaluated for the signal in the Wigner-Ville and Continuous Wavelet Transformation (CWT) domain. RESULTS: This Renyi's entropy gives lower 'p' value for scalogram than Wigner-Ville distribution and also, the contours of scalogram visually show the features of the diseases. And in the time-frequency analysis, the Renyi's entropy gives better result for scalogram than the Wigner-Ville distribution. CONCLUSION: Spatial filling index and Renyi's entropy has distinct regions for various diseases with an accuracy of more than 95%. PMID- 15361255 TI - Good physicians from the perspective of their patients. AB - BACKGROUND: It is not currently known what is the patient's viewpoint of a "good" physician. We set out to define patient's priorities regarding different physician's attributes in 3 domains important in medical care. METHODS: Patients hospitalized or attending clinics at a large teaching hospital selected the 4 attributes that they considered most important out of 21 listed arbitrarily in a questionnaire. The questionnaire included 7 items each in the domains of patient autonomy, professional expertise and humanism. RESULTS: Participating patients (n = 445, mean age 57.5 +/- 16 years) selected professional expertise (50%), physician's patience and attentiveness (38% and 30%, respectively), and informing the patient, representing the patient's interests, being truthful and respecting patient's preferences (25-36% each) as the most essential attributes. Patient's selections were not significantly influenced by different demographic or clinical background. Selections of attributes in the domain of patient's autonomy were significantly more frequent and this was the preferred domain for 31% and as important as another domain for 16%--significantly more than the domain of professional expertise (P = 0.008), and much more than the domain of humanism and support (P < 0.0005). CONCLUSIONS: Patients studied want their physicians to be highly professional and expert clinicians and show humaneness and support, but their first priority is for the physician to respect their autonomy. PMID- 15361256 TI - Screening for Parkinson's disease with response time batteries: a pilot study. AB - BACKGROUND: Although significant response time deficits (both reaction time and movement time) have been identified in numerous studies of patients with Parkinson's disease (PD), few attempts have been made to evaluate the use of these measures in screening for PD. METHODS: Receiver operator characteristic curves were used to identify cutoff scores for a unit-weighted composite of two choice response tasks in a sample of 40 patients and 40 healthy participants. These scores were then cross-validated in an independent sample of 20 patients and 20 healthy participants. RESULTS: The unit-weighted movement time composite demonstrated high sensitivity (90%) and specificity (90%) in the identification of PD. Movement time was also significantly correlated (r = 0.59, p < 0.025) with the motor score of the Unified Parkinson's Disease Rating Scale (UPDRS). CONCLUSIONS: Measures of chronometric speed, assessed without the use of biomechanically complex movements, have a potential role in screening for PD. Furthermore, the significant correlation between movement time and UPDRS motor score suggests that movement time may be useful in the quantification of PD severity. PMID- 15361257 TI - Feasibility of a patient decision aid regarding disclosure of personal health information: qualitative evaluation of the Health Care Information Directive. AB - BACKGROUND: Concerns regarding the privacy of health information are escalating owing both to the growing use of information technology to store and exchange data and to the increasing demand on the part of patients to control the use of their medical records. The objective of this study was to evaluate the Health Care Information Directive (HCID), a recently-developed patient decision aid that aims to delineate the level of health information an individual is willing to share. METHODS: We convened a series of four focus group meetings with several communities in a large Canadian city. A total of 28 men and women participated, representing health care consumer advocates, urban professionals, senior citizens, and immigrants who speak English as a second language. Data were analysed using qualitative methods. RESULTS: Participants lacked substantial knowledge regarding the fate and uses of personal health information. They expressed mistrust concerning how their information will be used and protected. Several suggestions were made towards customizing the use of data according to specific needs rather than broad and full access to their charts. Furthermore, despite concern regarding the implementation of a tool like the HCID, participants were hopeful that a refined instrument could contribute to the improved regulation of health information. CONCLUSION: This study indicated poor knowledge concerning the uses of personal health information, distrust concerning security provisions, and cautious support for a patient decision aid such as the HCID to improve control over health data. PMID- 15361258 TI - Survey of attitudes, materials and methods employed in endodontic treatment by general dental practitioners in North Jordan. AB - BACKGROUND: General dental practitioners provide the majority of endodontic treatment in Jordan. The aim of this study was to gather information on the methods, materials and attitudes employed in root canal treatment by dentists in North Jordan, in order to evaluate and improve the quality of current practice. METHODS: A questionnaire was posted to all registered general dental practitioners working in private practice in Irbid Governate in North Jordan (n = 181). The questionnaire included information on methods, materials and techniques used in endodontic treatment. RESULTS: Reply rate was 72% (n = 131). The results demonstrated that only five dentists used rubber dam occasionally and not routinely. The majority used cotton rolls for isolation solely or in combination with a high volume saliva ejector (n = 116). The most widely used irrigants were sodium hypochlorite and hydrogen peroxide, which were used by 32.9% (n = 43) and 33.6% (n = 44) of the respondents, respectively. Forty eight percent of the respondents (n = 61) used the cold lateral condensation technique for canal obturation, 31.3% (n = 41) used single cone, 9.9% (n = 13) used vertical condensation and 12.2% (n = 16) used paste or cement only for the obturation. The majority used zinc oxide eugenol as a sealer (72.5%). All, but one, respondents used hand instruments for canal preparation and the technique of choice was step back (52.7%). More than 50% (n = 70) of the dentists took one radiograph for determining the working length, whilst 22.9% (n = 30) did not take any radiograph at all. Most practitioners performed treatment in three visits for teeth with two or more root canals, and in two visits for teeth with a single root canal. CONCLUSIONS: This study indicates that dentists practicing in North Jordan do not comply with international quality standards and do not use recently introduced techniques. Many clinicians never take a radiograph for determining the working length and never used rubber dam or intra-canal medicaments. PMID- 15361259 TI - B-Raf specific antibody responses in melanoma patients. AB - BACKGROUND: Mutations of the BRAF gene are the most common genetic alteration in melanoma. Moreover, BRAF mutations are already present in benign nevi. Being overexpressed and mutated, B-Raf is a potential target for the immune system and as this mutation seems to be an early event, a humoral immune response against this antigen might serve as a diagnostic tool for detection of high risk patients. METHODS: 372 sera of 148 stage IV melanoma patients and 119 sera of non melanoma patients were screened for B-Raf, B-Raf V599E and C-Raf specific antibodies by an ELISA assay. Sera were screened for specific total Ig and for IgG. Serum titers were compared with a two tailed Mann-Whitney U test. Sera with titers of 1:300 or higher were termed positive and groups were compared with a two tailed Fisher's exact test. RESULTS: B-Raf specific antibodies recognizing both B-Raf and B-Raf V599E were detected in 8.9% of the sera of melanoma patients and in 2,5% of the control group. Raf specific IgG was detected in some patients at very low levels. B-Raf specific antibody responses did not correlate with clinical parameters but in some cases, B-Raf antibodies emerged during disease progression. CONCLUSION: These findings imply that B-Raf is immunogenic in melanoma patients and that it might serve as a potential target for immunotherapy. However, B-Raf specific antibodies emerge at rather late stages of melanoma progression and are present only with a low frequency indicating that spontaneous B-Raf specific antibodies are not an early marker for melanoma, but rather may serve as a therapeutic target. PMID- 15361260 TI - Frequency of myasthenic crisis in relation to thymectomy in generalized myasthenia gravis: a 17-year experience. AB - BACKGROUND: Myasthenic crisis is the most serious life-threatening event in generalized myasthenia gravis (MG) patients. The objective of this study was to assess the long-term impact of thymectomy on rate and severity of these attacks in Iranian patients. METHODS: We reviewed the clinical records from 272 myasthenic patients diagnosed and treated in our neurology clinic during 1985 to 2002. Fifty-three patients were excluded because of unconfirmed diagnosis, ocular form of MG, contraindication to surgery, concomitant diseases and loss to follow up. The Osserman classification was used to assess the initial severity of the disease. Frequency and severity of the attacks were compared between two groups with appropriate statistical tests according to the nature of variables. Multivariate logistic regression analysis was used to assess the predictors of myasthenic crisis in the group of patients without thymoma. RESULTS: 110 patients were in thymectomy group and the other 109 patients were on medical therapy. These two groups had no significant differences with respect to age at onset, gender, Osserman score in baseline and follow up period. 62 patients (28.3% of all 219 patients) had reported 89 attacks of myasthenic crisis. 20 patients of 62 (32%) were in thymectomy group and 42 (68%) were in the other group. There was significant difference between the two groups in number of patients with crisis (P = 0.001; odds ratio = 2.8 with 95% CI of 1.5 to 5.2). In addition, these attacks were more severe in group of non-thymectomized patients as the duration of ICU admission was longer and they needed more ventilatory support during their attacks. Regression model showed thymectomy and lower age at onset as two predictors of decrement in myasthenic crisis rate in non-thymomatous MG patients. CONCLUSIONS: It is suggested that frequency and severity of myasthenic attacks as important endpoints in evaluation of MG patients. Thymectomy seems to have a preventive role on rate and severity of these attacks. PMID- 15361261 TI - What is the impact of the ACE gene insertion/deletion (I/D) polymorphism on the clinical effectiveness and adverse events of ACE inhibitors?--Protocol of a systematic review. AB - BACKGROUND: The Angiotensin Converting Enzyme (ACE) insertion/deletion (I/D) polymorphism has received much attention in pharmacogenetic research because observed variations in response to ACE inhibitors might be associated with this polymorphism. Pharmacogenetic testing raises the hope to individualise ACE inhibitor therapy in order to optimise its effectiveness and to reduce adverse effects for genetically different subgroups. However, the extent of its effect modification in patients treated with ACE inhibitors remains inconclusive. Therefore our objective is to quantify the effect modification of the insertion/deletion polymorphism of the angiotensin converting enzyme gene on any surrogate and clinically relevant parameters in patients with cardiovascular diseases, diabetes, renal transplantation and/or renal failure. METHODS: Systematic Review. We will perform literature searches in six electronic databases to identify randomised controlled trials comparing the effectiveness and occurrence of adverse events of ACE inhibitor therapy against placebo or any active treatment stratified by the I/D gene polymorphism. In addition, authors of trials, experts in pharmacogenetics and pharmaceutical companies will be contacted for further published or unpublished data. Hand searching will be accomplished by reviewing the reference lists of all included studies. The methodological quality of included papers will be assessed. Data analyses will be performed in clinically and methodologically cogent subgroups. The results of the quantitative assessment will be pooled statistically where appropriate to produce an estimate of the differences in the effect of ACE inhibitors observed between the three ACE genotypes. DISCUSSION: This protocol describes a strategy to quantify the effect modification of the ACE polymorphism on ACE inhibitors in relevant clinical domains using meta-epidemiological research methods. The results may provide evidence for the usefulness of pharmacogenetic testing for individualised ACE inhibitor therapy. PMID- 15361262 TI - Plasticity of histamine H3 receptor expression and binding in the vestibular nuclei after labyrinthectomy in rat. AB - BACKGROUND: In rat, deafferentation of one labyrinth (unilateral labyrinthectomy) results in a characteristic syndrome of ocular and motor postural disorders (e.g., barrel rotation, circling behavior, and spontaneous nystagmus). Behavioral recovery (e.g., diminished symptoms), encompassing 1 week after unilateral labyrinthectomy, has been termed vestibular compensation. Evidence suggesting that the histamine H3 receptor plays a key role in vestibular compensation comes from studies indicating that betahistine, a histamine-like drug that acts as both a partial histamine H1 receptor agonist and an H3 receptor antagonist, can accelerate the process of vestibular compensation. RESULTS: Expression levels for histamine H3 receptor (total) as well as three isoforms which display variable lengths of the third intracellular loop of the receptor were analyzed using in situ hybridization on brain sections containing the rat medial vestibular nucleus after unilateral labyrinthectomy. We compared these expression levels to H3 receptor binding densities. Total H3 receptor mRNA levels (detected by oligo probe H3X) as well as mRNA levels of the three receptor isoforms studied (detected by oligo probes H3A, H3B, and H3C) showed a pattern of increase, which was bilaterally significant at 24 h post-lesion for both H3X and H3C, followed by significant bilateral decreases in medial vestibular nuclei occurring 48 h (H3X and H3B) and 1 week post-lesion (H3A, H3B, and H3C). Expression levels of H3B was an exception to the forementioned pattern with significant decreases already detected at 24 h post-lesion. Coinciding with the decreasing trends in H3 receptor mRNA levels was an observed increase in H3 receptor binding densities occurring in the ipsilateral medial vestibular nuclei 48 h post-lesion. CONCLUSION: Progressive recovery of the resting discharge of the deafferentated medial vestibular nuclei neurons results in functional restoration of the static postural and occulomotor deficits, usually occurring within a time frame of 48 hours in rats. Our data suggests that the H3 receptor may be an essential part of pre-synaptic mechanisms required for reestablishing resting activities 48 h after unilateral labyrinthectomy. PMID- 15361263 TI - Obesity among outpatients with major depressive disorder. AB - Studies focusing on the prevalence of obesity in Major Depressive Disorder (MDD), or the impact of excess body fat on the treatment of MDD are lacking. The aim of the present work is to systematically study obesity in MDD outpatients. A total of 369 MDD outpatients enrolled in an 8-wk trial of 20 mg fluoxetine had height and weight measured at baseline. We then examined: (1) the prevalence of being overweight or obese, (2) the relationship between obesity and a number of demographic and clinical variables, and, (3) the relationship between relative body weight and obesity with clinical response. We found that more than 50% of patients were overweight [body mass index (BMI) > or =2 5 kg/m2], while 20% were obese (BMI > or = 30 kg/m2). Obese patients presented with worse somatic well being scores than non-obese MDD patients, but they did not differ with respect to depression severity, anxiety, somatic complaints, hopelessness or hostility. Greater relative body weight, but not obesity, predicted non-response. In conclusion, greater relative body weight was found to place MDD outpatients at risk for fluoxetine resistance. PMID- 15361265 TI - Primary thyroid lymphoma associated with Graves' disease. AB - We report herein a case of thyroid mucosa-associated lymphoid tissue (MALT) lymphoma in a patient receiving antithyroid drug therapy for Graves' disease. A 75-year-old woman first presented with finger tremor and was diagnosed with Graves' disease on the basis of clinical and laboratory findings. Three years later, she presented with rapid and painless enlargement of the thyroid. Ultrasonography revealed a circumscribed hypoechoic area bilaterally in each lobe of the thyroid, and fine-needle aspiration biopsy showed diffuse monotonous infiltration of small- to medium-sized atypical lymphoid cells. (67)Ga scintigraphy was positive exclusively in the thyroid. After total thyroidectomy, the patient received radiation therapy for treatment of stage IE primary thyroid lymphoma. Results of histological examination, immunohistochemical analysis, and flow cytometric analysis confirmed MALT lymphoma. To our knowledge, there have been few published reports of primary thyroid lymphoma associated with Graves' disease. Our experience with this case, though rare, indicates that an enlarged thyroid in cases of Graves' disease should be examined carefully for primary thyroid lymphoma. PMID- 15361266 TI - Thyroglossal duct carcinoma in children: case presentation and review of the literature. AB - Carcinoma within the thyroglossal duct (TGDCa) is a very rare pediatric tumor. This report presents the case of a 15-year-old girl with TGDCa, reviews the previously published pediatric cases of this tumor, and provides diagnostic and therapeutic considerations in TGDCa in children. Twenty one cases of TGDCa have been reported, 12 of them in females. Mean age at presentation was approximately 13 years for females and approximately 12 years for males. The duration of a midcervical mass prior to the diagnosis of TGDCa varied from 3 weeks to 8 years. The size of the neck mass at presentation averaged 2 cm-4 cm. All masses were asymptomatic, and the diagnosis of TGDCa was incidental following surgery. All pediatric cases of TGDCa reported thus far were of the papillary type, except for 3 patients who presented with a mixed papillary-follicular carcinoma. Capsular invasion was detected in 10 (45%) patients. Local invasive disease was found in 5 (23%) patients, but all removed thyroid glands (12) were free of carcinoma. One patient had lung metastases. Thyroidectomy with subsequent radioiodine ablation was the treatment of choice in the majority of the cases. The reported prognosis for TGDCa in children was favorable, with only one reported death. PMID- 15361267 TI - Aspergillus fumigatus abscesses of the thyroid with obstruction of the esophagus. AB - Thyroid invasion by Aspergillus spp. can occur with invasive aspergillosis, although it is rarely diagnosed antemortem. We describe a case of multiple thyroid abscesses from A. fumigatus that caused esophageal obstruction in a patient with myelodysplasia. Despite aggressive antifungal treatment, the outcome was rapidly fatal. PMID- 15361268 TI - Thyroid hormone and parathyroid hormone competing to maintain calcium levels in the presence of vitamin D deficiency. AB - Weight loss and bone disease in the elderly are very often attributed to malignancy. Rarely, benign treatable conditions may be overlooked. Thyrotoxicosis, a benign treatable condition, needs to be excluded in such patients. The diagnosis may be delayed, since the symptoms are often subtle, and secondary complications including bone disease (osteoporosis) are therefore more frequent at the time of presentation. The case presented here illustrates this well, and also highlights the value of measuring vitamin D levels in such patients. The most interesting aspect of this case was the reciprocal relationship between thyroxine and parathyroid hormone observed in maintaining calcium homeostasis in this thyrotoxic patient with low vitamin D levels. PMID- 15361269 TI - Two lumps in the neck. PMID- 15361270 TI - Angiogenesis in thyroid cancers. PMID- 15361272 TI - CanMEDs roles and obstetrics and gynaecology: the time is now. PMID- 15361273 TI - Human papilloma virus testing knowledge and attitudes among women attending colposcopy clinic with ASCUS/LGSIL pap smears. AB - OBJECTIVE: To study women's knowledge regarding the role of human papilloma virus (HPV) in cervical intraepithelial neoplasia and their attitudes toward the integration of HPV testing as part of routine follow-up of atypical squamous cell of uncertain significance/low-grade squamous intraepithelial lesion (ASCUS/LGSIL) abnormalities. METHODS: Over a 12-month period, all women attending the University of Ottawa colposcopy clinic for evaluation and follow-up of ASCUS/LGSIL Pap smears were recruited. Demographic data included age, nature of the Pap smear abnormality, gravidity, parity, occupation and education level, smoking history, previous history of abnormal smears, colposcopic examination and treatment, and current method of contraception. The women were asked to rate their level of concern over their Pap smear abnormality, from 0 (not concerned) to 10 (very concerned). Women's knowledge regarding the role of HPV in cervical intraepithelial neoplasia and the rationale behind the use of HPV testing was assessed by the clinic nurse as being minimal, moderate, or good, as defined by pre-specified criteria. Upon explanation by the nurses of the results of the recent ALTS (ASCUS/LGSIL Triage Study) trial, the women were asked to state whether they preferred to continue with regular colposcopic surveillance every 6 months, or to use the results of the HPV test, if negative, to reduce the number of colposcopy examinations to one annually. Descriptive statistics and logistic regression analysis were used to identify significant demographic factors associated with the women's preference for incorporation of HPV testing in their follow-up. All P values less than.10 were considered to be statistically significant, due to the exploratory nature of the study. RESULTS: Of the 100 women who participated in the study, 42% presented with ASCUS. The mean age (+/- SD) of the women was 33.63 +/- 11.25 years (range, 18-75 years); 66% were office workers with at least a community college degree, 86% reported previous abnormal Pap smears, and 67% had previously been seen for colposcopy. Fifty-eight percent of the women rated their concern level as being 6 or more, while 15% ranked their concerns as maximal at 10. In terms of knowledge about HPV, 75% of the women had no or very minimal knowledge of the role of HPV in cervical intraepithelial neoplasia. With regard to HPV testing, 84% of the study group had either never heard of the test or had only a minimal knowledge of HPV testing. After being informed of the ALTS results, 64% of the women chose to use HPV testing to help in triaging the needs for frequent colposcopy. Logistic regression modelling showed that a college level education (odds ratio [OR], 2.27; 95% confidence interval [CI], 0.95-;5.45; P =.06) and history of previous treatment for abnormal Pap smears (OR, 3.31; CI, 0.88- 12.46; P =.07) were closely associated with the adoption of HPV testing in clinical management. CONCLUSION: There exists a significant lack of knowledge about HPV and its role in the pathogenesis of cervical intraepithelial neoplasia. Women who have received previous treatments for cervical intraepithelial neoplasia and those with college-level educations were more likely to adopt this new technology as part of their care. PMID- 15361274 TI - A comparison of endometrial cancer outcomes in Ontario. AB - OBJECTIVE: To compare endometrial cancer treatment strategies and outcomes across the province of Ontario, Canada. METHODS: A retrospective cohort study was conducted of 195 women diagnosed with endometrial cancer in Ontario between 1996 and 1998, as a sample of the population. The women's charts were randomly selected by the medical records departments at 5 tertiary care centres in Ontario. The outcomes measured included 5-year overall survival (OS) and disease free survival (DFS), use of adjuvant radiotherapy, treatment complications, and prognostic factors for survival. RESULTS: The 2 main treatment strategies were (1) total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH BSO) and (2) surgical staging (defined as TAH BSO and pelvic lymph node dissection with or without cytology, peritoneal biopsies, and omentectomy). Surgical staging rates across the province ranged from 0% to 88%. Stratified survival analysis revealed a significant difference in OS among centres (log rank P =.039). Crude survival analysis revealed no difference in 5-year OS or DFS between the 2 treatment strategies. The Cox proportional hazards model identified advanced stage of tumour as being the most predictive factor of DFS, and the woman's age at diagnosis and tumour grade as predictive of OS. DISCUSSION: There was a significant difference in 5-year OS among the 5 tertiary care centres. There was no significant difference between surgical staging and TAH BSO with respect to 5 year DFS or OS. CONCLUSION: As there were significant differences in the treatment of endometrial carcinoma and OS across the province, a population-based study of endometrial cancer treatment strategies and outcomes is required. PMID- 15361275 TI - Trends in concurrent maternal and perinatal deaths at a teaching hospital in Ghana: the facts and prevention strategies. AB - OBJECTIVES: To determine the trend of concurrent maternal and perinatal mortality at the Korle-Bu Teaching Hospital (KBTH), Ghana, and to propose measures for its prevention. METHODS: A retrospective study, from January 1995 to December 2002, of all concurrent maternal and perinatal deaths in which the woman was 28 weeks' gestation or more (or, if gestational age was not known, the baby weighed 1000 g or more) and died either undelivered or in the perinatal period (within 1 week of delivery) at the KBTH. RESULTS: Over the 8-year study period, there was a total of 93 622 deliveries at the KBTH with 108 concurrent maternal and perinatal mortalities, giving a ratio of 115.4 concurrent maternal and perinatal deaths per 100 000 deliveries. More than 80% of the mothers who died had little or no formal education. Of the 108 mothers, 22 died undelivered. The leading cause of death was a medical condition in pregnancy along with eclampsia/gestational hypertension. Of the 86 delivered mothers, the leading cause of concurrent death was a medical condition in pregnancy. Approximately two-thirds (72/108) of the perinatal deaths were stillbirths. Over the study period, there was a rising trend of the obstetric disaster of losing both mother and baby. CONCLUSIONS: There is a rising trend of concurrent maternal and perinatal mortality at the KBTH. It is suggested that a regular antenatal clinic be established with both an internist and obstetrician to jointly see and manage women with medical problems. There is a need for improved and adequate resources to improve outcomes for both mother and baby. A waiver of user fees for maternity services may be one way to improve access for needy and at-risk mothers. Concurrent maternal and perinatal death is the latest negative reproductive health index of the deteriorating socioeconomic situation in developing countries and needs to be tackled decisively. PMID- 15361276 TI - Surgical management of severe obstetric hemorrhage: experience with an obstetric hemorrhage equipment tray. AB - BACKGROUND: Hemorrhage remains a leading cause of severe maternal morbidity and death. In addition to the appropriate use of oxytocic agents for uterine atony, surgical techniques, including uterine tamponade, major vessel ligation, compression sutures, and hysterectomy, may be required. On the rare occasions they are needed, the instruments and equipment required for these surgical techniques may not be readily available. OBJECTIVE: To record our experience with an obstetric hemorrhage equipment tray for surgical management of severe obstetric hemorrhage. METHODS: A severe obstetric hemorrhage equipment tray was established and included packing (5-yard roll) and balloon device for uterine tamponade, straight (10 cm) eyed-needles and large curved eyed-needles for use with No. 1 suture, 3 Heaney vaginal retractors, 4 sponge forceps, and diagrams and instructions for the various types of compression sutures and tamponade techniques. RESULTS: Of the 4400 deliveries that occurred at 1 tertiary maternity hospital during 2002, the obstetric hemorrhage tray was used on 9 occasions: 3 Caesarean sections with placenta previa (uterine tamponade used in 2 cases, compression sutures in 1); 2 Caesarean sections with uterine atony (compression sutures); 1 Caesarean section with placenta previa accreta (major vessel ligation and compression sutures); and 3 vaginal deliveries (suturing of cervical and vaginal lacerations in 2 of the cases, uterine tamponade used in the third case). In all cases, hysterectomy was avoided. CONCLUSION: The ready availability of an obstetric hemorrhage equipment tray on the labour ward facilitates prompt surgical management of severe obstetric hemorrhage, and may reduce the need for blood transfusion and hysterectomy. PMID- 15361277 TI - Iron bioavailability in prenatal multivitamin supplements with separated and combined iron and calcium. AB - OBJECTIVE: To compare iron absorption of a prenatal multivitamin supplement containing both iron and calcium (HICA) to that of another multivitamin containing a lower iron dose and no calcium (LI). METHODS: In a crossover study, serum iron was measured in 12 healthy women administered HICA and LI separately on 2 different occasions. Blood samples were taken at 0, 1, 2, 3, 4, 6, and 8 hours after administration of each supplement. RESULTS: The values of the area under the concentration-time curve (AUC) were not significantly different between LI (79.1 +/- 36.0 micro M*h) and HICA (91.4 +/- 50.4 micro M*h) (P =.37). After standardizing the AUC for dose, the relative absorption over the 8-hour time period was significantly higher for LI (2.3 +/-.0 micro M*h/mg) than for HICA (1.5 +/- 0.8 micro M*h/mg) (P =.021). CONCLUSION: The absorption of iron from a low-iron-containing supplement was similar to that from a supplement with almost twice the amount of iron, due possibly to the exclusion of calcium in the LI product. Thus, while offering similar amounts of iron, the LI supplement may be better tolerated by women who are sensitive to iron-induced adverse effects. PMID- 15361278 TI - Perceived educational benefits of objective structured clinical examination (OSCE) development and implementation by resident learners. AB - OBJECTIVE: To assess the perceived learning outcomes from the various roles involved in the development and implementation of an objective structured clinical examination (OSCE) by residents for residents. METHODS: Final-year residents in Obstetrics and Gynaecology at the University of Toronto created OSCE stations in preparation for their certification examination. They evaluated their experience using a survey designed to assess the learning effectiveness of the roles played in an OSCE. Residents were asked to compare resident-created stations with faculty-created stations. RESULTS: The students found every aspect of OSCE development to be of educational benefit. Residents rated the candidate role as more beneficial than other roles. Residents perceived the benefits of the OSCE sessions to be greater than equivalent lengths of time spent in traditional group study sessions. CONCLUSIONS: A self-directed learning approach, based on OSCE development and implementation, shows promise as a learning aid at the senior residency level. We suggest a controlled trial designed to objectively measure outcomes of this learner-centred approach. PMID- 15361279 TI - The use of selective serotonin reuptake inhibitors in pregnancy. AB - OBJECTIVE: To provide an update of literature on the safety of using selective serotonin reuptake inhibitors (SSRIs) during pregnancy. METHODS: MEDLINE was searched for English-language papers published from 1985 to 2003 on human studies of SSRIs, using the key words "serotonin reuptake inhibitors," "citalopram," "fluoxetine," "fluvoxamine," "paroxetine," and "sertraline." RESULTS: The literature search yielded 12 338 publications. Previous studies on the safety of SSRIs in pregnancy were often based on small samples from medical centres, with heterogeneous design and outcome ascertainment methods, and had yielded inconsistent results. Consequently, the management of pregnant women with depression poses challenges to clinicians who are hesitant to prescribe anti depression drugs, including SSRIs, because of concern about potential risks to the fetuses. Failure to adequately treat maternal depression can lead to progressively worsening depression that greatly compromises maternal-fetal health and can impair bonding and childcare in the postpartum period. CONCLUSIONS: Because of the uncertainty regarding the safety of SSRI use during pregnancy, consultation with specialists experienced in treating depression may be helpful when treating pregnant women with SSRIs. Large-scale, population-based studies to comprehensively assess the safety of SSRIs in pregnancy are needed. PMID- 15361280 TI - Cystic teratoma-provoked peritonitis after pregnancy termination. AB - BACKGROUND: Cystic teratoma is an unusual cause of postabortal pelvic peritonitis. CASE: A healthy 22-year-old woman underwent outpatient first trimester vacuum pregnancy termination, followed by a 3-day course of oral doxycycline. On the thirteenth day after pregnancy termination, she presented with fever, an acute abdomen, and pelvic pain. The woman underwent re-aspiration of her uterus, which yielded scanty tissue. She then underwent pelvic ultrasound and computerized tomogram, both of which revealed a posterior pelvic mass, presumed to be an abscess. After 5 days of broad-spectrum antibiotics had failed to alleviate her fever and pain, culdocentesis was performed yielding 30 cm3 of pus, hair, and fat. The woman underwent laparotomy, which revealed a 10 x 8 x 5 cm multiloculated, infected benign cystic teratoma with extensive capsular adhesions. CONCLUSION: If postoperative infection is to develop after pregnancy termination, signs will usually appear within the first week. Onset of pelvic peritonitis more than 1 week after uneventful conclusion of a pregnancy event should raise suspicion of either non-gynaecologic causes or, as in this case, a pelvic neoplasm. PMID- 15361281 TI - The prevention of early-onset neonatal group B streptococcal disease. AB - OBJECTIVE: To review the evidence in the literature and to provide recommendations on the management of pregnant women in labour for the prevention of early-onset neonatal group B streptococcal (GBS) disease. OUTCOMES: Maternal outcomes evaluated included exposure to antibiotics in pregnancy and labour and complications related to antibiotic use. Neonatal outcomes of rates of early onset group B streptococcal infections are evaluated. EVIDENCE: A review of the literature through MEDLINE from January 1980 to December 2003, relating to neonatal group B streptococcal infection and a review of the Centers for Disease Control and Prevention recommendations. VALUES: The evidence obtained was reviewed and evaluated by the Infectious Diseases Committee of the Society of Obstetricians and Gynaecologists of Canada (SOGC) under the leadership of the principal authors, and recommendations were made according to guidelines developed by the Canadian Task Force on the Periodic Health Exam. RECOMMENDATIONS: 1. Offer all women screening for group B streptococcal disease at 35 to 37 weeks' gestation with culture done from one swab first to the vagina then to the rectal area. (II-1)2. Treat the following women intrapartum at time of labour or rupture of membranes with IV antibiotics: -all women positive by GBS culture screening done at 35 to 37 weeks (II-2) - any women with an infant previously infected with GBS (II-3) - any women with documented GBS bacteriuria (regardless of level of colony-forming units per mL) in this pregnancy (II-2) 3. Treat women at less than 37 weeks' gestation with IV antibiotics unless there has been a negative GBS vaginal/rectal swab culture within 5 weeks. (II-3) 4. Treat women with intrapartum fever with IV antibiotics (i.e., chorioamnionitis must be treated, but broader spectrum antibiotics would be advised). (II-2) 5. If a woman is GBS-positive by culture screening or by history of bacteriuria, with prelabour rupture of membranes at term, treat with GBS antibiotic prophylaxis and initiate induction of labour with IV oxytocin (II-1) 6. If GBS culture result is unknown and the woman has ruptured membranes at term for greater than 18 hours, treat with GBS antibiotic prophylaxis. (II-2) PMID- 15361282 TI - Chemokine-like factor 1, a novel cytokine, contributes to airway damage, remodeling and pulmonary fibrosis. AB - BACKGROUND: Chemokine-like factor 1 (CKLF1) was recently identified as a novel cytokine. The full-length CKLF1 cDNA contains 530 bp encoding 99 amino acid residues with a CC motif similar to that of other CC family chemokines. Recombinant CKLF1 exhibits chemotactic activity on leucocytes and stimulates proliferation of murine skeletal muscle cells. We questioned whether CKLF1 could be involved in the pathogenesis of inflammation and proliferation in the lung. Therefore we used efficient in vivo gene delivery method to investigate the biological effect of CKLF1 in the murine lung. METHODS: CKLF1-expressing plasmid, pCDI-CKLF1, was constructed and injected into the skeletal muscles followed by electroporation. Lung tissues were obtained at the end of week 1, 2, 3 and 4 respectively after injection. The pathological changes in the lungs were observed by light microscope. RESULTS: A single intramuscular injection of CKLF1 plasmid DNA into BALB/c mice caused dramatic pathological changes in the lungs of treated mice. These changes included peribronchial leukocyte infiltration, epithelial shedding, collagen deposition, proliferation of bronchial smooth muscle cells and fibrosis of the lung. CONCLUSIONS: The sustained morphological abnormalities of the bronchial and bronchiolar wall, the acute pneumonitis and interstitial pulmonary fibrosis induced by CKLF1 were similar to phenomena observed in chronic persistent asthma, acute respiratory distress syndrome and severe acute respiratory syndrome. These data suggest that CKLF1 may play an important role in the pathogenesis of these important diseases and the study also implies that gene electro-transfer in vivo could serve as a valuable approach for evaluating the function of a novel gene in animals. PMID- 15361283 TI - Use of B-type natriuretic peptide in evaluation of early percutaneous coronary intervention in patients with acute coronary syndrome. AB - BACKGROUND: Previous studies showed that blood B-type natriuretic peptide (BNP) level could predict the prognosis of acute coronary syndromes (ACS). This study investigated the evaluation value of circulating BNP for early percutaneous coronary intervention (PCI) in patients with ACS. METHODS: Nine hundred and sixty consecutive patients with ACS were enrolled. Circulating BNP level was measured when each patient arrived at the emergency room. All patients underwent PCI in 90 minutes in spite of contraindication. Cardiac events (death from any cause, heart failure, and recurrence of acute myocardial infarction or ACS) were recorded during follow-up. RESULTS: In patients with BNP > or = 80 pg/ml, mortality from all causes within 1 month and 6 months in those underwent delayed PCI (> or = 6 hours) was significantly higher than those received early PCI (< 6 hours) (9.53% vs 3.49%, P = 0.027; 13.61% vs 5.24%, P = 0.010, respectively). Similarly, the incidence rate of heart failure in delayed PCI patients was significantly higher than those received early PCI within 1 month and 6 months (12.93% vs 4.66%, P = 0.008; 14.97% vs 6.98%, P = 0.021, respectively). The recurrence rate of acute myocardial infarction or ACS, however, was not significantly different between early PCI and delayed PCI patients in group BNP > or = 80 pg/ml. In patients with BNP < 80 pg/ml, no significant difference was observed between early PCI and delayed PCI patients with any of the above cardiac events within 1 month or 6 months. CONCLUSION: While early level of circulating BNP > or = 80 pg/ml, the incidence of mortality and heart failure, but not recurrence of acute myocardial infarction, is significantly reduced in patients with ACS provided by early PCI. PMID- 15361284 TI - Crosstalk between ERK1/2 and STAT3 in the modulation of cardiomyocyte hypertrophy induced by cardiotrophin-1. AB - BACKGROUND: The Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway and the extracellular signal-regulated kinases 1/2 (ERK1/2) pathway are the two major independent signal transduction pathways. However, it has recently been found that STAT3 may be negatively regulated by ERK1/2 in gp130 dependent signaling. Cardiotrophin-1 (CT-1), a potent novel hypertrophic cytokine, depends on gp130 to induce signaling and depends on STAT3 to exert hypertrophic effect. In this study, we examined whether STAT3 activity was negatively regulated by ERK1/2 during CT-1-induced signaling in rat cardiomyocytes and, if so, whether such crosstalk interfered with the hypertrophic effect of CT-1 and, furthermore, whether the mechanism underlying the crosstalk involved phosphorylation of serine 727 (S727) in STAT3. METHODS: The activities of ERK1/2 and STAT3 were assessed by in-gel kinase assay and Western blot analysis, respectively. The role of S727 phosphorylation in the crosstalk between ERK1/2 and STAT3 was determined by a transient transfection study using a STAT3S727A mutant. Cardiomyocyte hypertrophy was evaluated by the cellular protein-to-DNA ratio and [(3)H]-leucine incorporation. RESULTS: CT-1 simultaneously activated both ERK1/2 and STAT3 in rat cardiomyocytes. Inhibition of ERK1/2 by U0126 resulted in an increase of CT-1-induced tyrosine phosphorylation of STAT3 and, consequently, the protein-to-DNA ratio and [(3)H] leucine incorporation. Transient transfection of the cells with STAT3S727A had no significant effect on CT-1-induced tyrosine phosphorylation of STAT3. CONCLUSIONS: STAT3 is activated by CT-1 in rat cardiomyocytes, but full activation is mitigated by the simultaneous activation of ERK1/2. The inhibition of ERK1/2 increases the activity of STAT3, which, in turn, enhances the hypertrophic effect of CT-1. The crosstalk between ERK1/2 and STAT3 is independent of the phosphorylation of the S727 in STAT3. Such crosstalk may contribute to the development of adequate cardiac hypertrophy. PMID- 15361285 TI - Rac1 regulates the release of Weibel-Palade Bodies in human aortic endothelial cells. AB - BACKGROUND: The release of Weibel-Palade Bodies (WPB) is a form of endothelial cell activation. But the signal transduction pathway leading to WPB release is not yet defined. We hypothesized that small G-protein rac1 and reactive oxygen species (ROS) mediate the ligand induced release of Weibel-Palade Bodies. METHODS: We tested this hypothesis by using wild-type and mutant adenoviral rac1 expression vectors, and by manipulating the production and destruction of superoxide and hydrogen peroxide in human aortic endothelial cells (HAEC). RESULTS: Thrombin (1.0 Unit, 30 min) induced the increase of WPB release by 3.7 fold in HAEC, and that H2O2 (0.1 mmol/L, 30 min) induced by 4.5-fold. These results correlated with thrombin-stimulated activation of rac-GTP binding activity by 3.5-fold, and increase of ROS production by 3.4-fold. The dominant negative adenoviral rac-N17 gene transfer dramatically inhibited the release of WPB by 64.2% (control) and 77.3% (thrombin-stimulation), and decreased ROS production by 65.5% (control) and 83.6% (thrombin-stimulation) compared with non infected cells, respectively. Anti-oxidants, catalase and N-acetyl-cysteine significantly decreased the release of WPB by 34% and 79% in control cells, and further decreased by 63.6% and 46.7% in rac-N17 transferred cells compared with non-infected cells. We also confirmed that rac1 was located upstream of ROS in the WPB release pathway. CONCLUSIONS: Small G-protein rac1 medicates ligand induced release of Weibel-Palade Bodies in human aortic endothelial cells, and the signal pathway of WPB release is a rac1-dependent ROS regulating mechanism. PMID- 15361286 TI - Relationship between COX-2 expression and clinicopathological features of colorectal cancers. AB - BACKGROUND: Cyclo-oxgenase 2 (COX-2) is involved in prostaglandin synthesis in central nervous system, and it also plays a role in human carcinogenesis. Our purpose of this study is to investigate the COX-2 expression in different development stages of colorectal cancer, and to discuss the relationship between the gene expression and clinicopathological features of the cancer. METHODS: COX 2 expression was examined by immunohistochemical staining in 76 surgical specimens of colorectal cancer (44 of advanced stage and 32 of early stage), thirty-three adenomas and 18 normal colonic mucosal tissues taken by endoscopic biopsy. Kaplan-Meier survival curves and Cox proportional hazards regression were used to evaluate the relation of COX-2 to prognosis. RESULTS: COX-2 expression, divided into 4 grades from "-" to "+++", is respectively 83.3%, 16.7%, 0% and 0% in normal colonic mucosal tissues; 12.1%, 42.4%, 36.4% and 9.1% in adenomas; 6.3%, 28.1%, 46.9% and 18.7% in early colorectal cancers (ECCs), and 6.8%, 20.5%, 18.2% and 54.5% in advanced colorectal cancers (CRCs). The differences in COX-2 expression between advanced CRCs and early colorectal cancers (ECCs) as well as between the advanced CRCs and adenomas were statistically significant (P < 0.01); but there was no significant difference between ECCs and adenomas. Kaplan-Meier survival analysis showed a significant difference in the survival curves between low high COX-2 groups (P < 0.05). Cox proportional hazards regression showed that COX-2 expression was related to poorer long-term outcome with a hazard ratio of 2.665 unadjusted for other variables (P < 0.05), and COX-2 expression was an independent risk factor of poor prognosis. CONCLUSIONS: COX-2 expression is gradually up-regulated in the development from normal epithelium to adenomas and from ECCs to advanced CRCs. Alhough the COX-2 protein can not be regarded as a tumor marker to diagnose CRCs early, COX-2 expression can be regarded as an independent risk factor of poor prognosis for postoperative patients with advanced CRCs. PMID- 15361287 TI - An intervention study to prevent gastric cancer by micro-selenium and large dose of allitridum. AB - BACKGROUND: People have more and more concerned about allitridum as studies have shown that taking more raw garlic associated with a lower risk for cancers of the alimentary system. In the present study, we tried to examine whether a large dose of allitridum and a microdose of selenium prevent gastric cancer. METHODS: A double-blind intervention study was performed on the participants aged (35 - 74) years, who had matched at least one of the following criteria: (1) a medical history of stomach disorder, (2) a family history of tumour, or (3) smoking and/or alcohol consumption. A total of 2,526 and 2,507 persons were randomly enrolled into intervention group and control group respectively from 288 natural villages of seven communities in Qixia County, Shandong Province, China. Each person of the intervention group orally took 200 mg synthetic allitridum every day and 100 microg selenium every other day for one month of each year during November 1989 to December 1991. At the same time, people in control group were given 2 placebo capsules containing corn oid with the identical appearance to that in the intervention group. RESULTS: For all subjects the large dose of allitridum was accepted and no harmful side effects were found during the study. In the first follow-up five years (1992 - 1997) after stopping the intervention, the morbidity rates of malignant tumours in the intervention group declined by 22%, in contrast to the control group, declined by 47.3%. After adjusting for age, gender, and other potential confounders, relative risks (RRs) for all tumours and gastric cancer of the whole population were 0.67 (95% CL: 0.43 - 1.03) and 0.48 (95% CL: 0.21 - 1.06), respectively, and for male group they were 0.51 (95% CL: 0.30 - 0.85) and 0.36 (95% CL: 0.14 - 0.92), respectively. No signigicantly protective effect was found for the female subgroup. CONCLUSION: The present study proves that large doses of allitridum and microdorse of selenium may effectively prevent gastric cancer, especially in men. PMID- 15361288 TI - Differential acetylcholine and choline concentrations in the cerebrospinal fluid of patients with Alzheimer's disease and vascular dementia. AB - BACKGROUND: An important aspect of Alzheimer's disease (AD) is loss or impairment of cholinergic neurons. It is controversial whether there is a similar cholinergic impairment and cerebral deficit of acetylcholine (ACh) in the case of vascular dementia (VD). The purpose of this study was to explore the levels of ACh and choline (Ch) in the cerebrospinal fluid (CSF) of patients with AD and VD, and their possible relationship with cognitive impairment. METHODS: Twenty-two AD patients, twenty-two VD patients, and twenty normal controls were recruited and scored with a Mini-Mental State Examination (MMSE). CSF concentrations of ACh and Ch were measured using high-performance liquid chromatography with an electrochemical detector (HPLC-ECD) and the results were then compared to cognitive status. RESULTS: ACh concentrations in CSF of AD patients [(10.7 +/- 5.1) nmol/L] and VD patients [(16.8 +/- 7.4) nmol/L] were both significantly lower than in controls [(34.5 +/- 9.0) nmol/L, t = 10.67, P < 0.001; t = 6.91, P < 0.001]. Both results correlated positively with MMSE scores (rs = 0.88 and rs = 0.85, respectively, P < 0.01). The CSF concentration of Ch was significantly higher in VD patients [(887.4 +/- 187.4) nmol/L] compared to AD patients [(627.6 +/- 145.1) nmol/L, t = 6.4, P < 0.001] and controls [(716.0 +/- 159.4) nmol/L, t = 4.2, P = 0.002]. CSF Ch concentration showed no difference between AD patients and normal controls, nor did it correlate with MMSE score in any of the three groups. CONCLUSIONS: The positive correlation between ACh deficit and cognitive impairment suggests that ACh is an important neurotransmitter for memory. The similar decrease in ACh concentration in AD and VD patients may imply a similar pathogenesis for the process of cognitive impairment involved in these two disorders. The elevated CSF levels of Ch in VD patients compared to AD patients may be useful diagnostically. Cholinesterase inhibitors may be helpful not only for AD patients, but also for VD patients. PMID- 15361289 TI - Changes of macrovascular endothelial ultrastructure and gene expression of endothelial nitric oxide synthase in diabetic rats. AB - BACKGROUND: The most intimidatory pathological changes in patients with DM are cardiovascular illnesses, which are the major causes of death in diabetic patients and are far more prevalent than in nondiabetics because of accelerated atherosclerosis. In this study, we tried to clarify the changes in macrovascular endothelial ultrastructure and in the gene expression of endothelial nitric oxide synthase (eNOS)mRNA in diabetic rats. METHODS: The study was conducted on 52 of 10-week old Sprague Dawley (SD) rats with body weight of (320 +/- 42) g. SD rats were divided into: experimental group treated with a single intraperitoneal injection of streptozotocin (STZ, 60 mg/kg), (male, n = 20, diabetes mellitus (DMM)); female, n = 12, diabetes mellitus female (DMF)) and control group (male, n = 10, diabetes mellitus male control (DMMC); female, n = 10, diabetes mellitus female control (DMFC)). Four weeks after treatment, half of the rats were sacrificed; the remainders were sacrificed ten weeks after treatment. One part of the abdominal aortic sample was stored under glutaraldehyde (volume fraction psiB = 2.5%). After the process of chemical fixation, chemical dehydration, drying and conductivity enhancement, all samples were observed and photographed using scanning electron microscopy (Leica-Stereoscan 260, England). The other part of the abdominal aortic sample was treated with liquid nitrogen and the expression of eNOSmRNA was assessed by semi-quantitative RT-PCR. RESULTS: The aortic lumen of both experimental groups adsorbed much more debris than that of either control group. The endothelial surfaces of diabetic rats were coarse, wrinkled and protuberant like fingers or villi. The vascular endothelial lesions of diabetic male rats were very distinct after 4 weeks, and as obvious as those at 10 weeks. The vascular endothelial lesions of diabetic female rats were not severe at 4 weeks and only became marked after 10 weeks. In both males and females, the abdominal aortic eNOSmRNA content of 4 weeks and 10 weeks diabetic rats was very significantly lower (P < 0.01) than that of controls. CONCLUSIONS: Aortic endothelial ultrastructure in DM rats is injured compared with controls. Abnormal changes of aortic endothelia in male DM rats are more obvious than those in females. Expression of abdominal aortic eNOSmRNA content of DM rats is significantly lower than that of controls. PMID- 15361290 TI - Inhibitory effect of retroviral vector containing anti-sense Smad4 gene on Ito cell line, LI90. AB - BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) exerts strong fibrogenic potential in culture-activated HSCs. Smad4 is a key intracellular mediator for the transforming growth factor-beta (TGF-beta) superfamily of growth factors. The aim of this study was to assess the effects of the antisense Smad4 gene on Ito cell line, LI90. METHODS: The recombinant retroviral vector pLXSN-Smad4 was constructed by cloning the rat antisense Smad4 cDNA into the retroviral vector pLXSN. Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad4 and pLXSN vectors into PA317 cells. Human hepatic stellate cells (HSCs) LI90 were infected with these retroviruses followed by selection with G418. The expression of Smad4 was detected by Northern and Western blots. Cell biological characteristics, including cell growth curve, 3H-TdR and 3H-proline uptake by HSCs and the production of extracellular matrix were assessed. RESULTS: mRNA and protein expressions of Smad4 in LI90 cells transfected with retrovirus containing the antisense Smad4 gene were much lower than those in LI90 cells transfected with empty vector or parental LI90 cells. Cells hypoexpressing the Smad4 gene exhibited a slower rate of growth, a lower uptake of 3H-TdR and 3H-proline (P < 0.01), and smaller production of th extracellular matrix, compared with parental LI90 cells and cells transfected with empty retrovirus. CONCLUSIONS: The antisense Smad4 gene can suppress the expression of the Smad4 gene, reduce endogenous production of Smad4 mRNA and protein, block TGF-beta1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix (ECM). Our results may provide a basis for the development of antifibrotic gene therapy. PMID- 15361291 TI - Anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways act as potent immunoregulatory cells in vitro and vivo. AB - BACKGROUND: This study was to evaluate whether anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro and prolong cardiac allograft survival after adoptive transfer. METHODS: Anergic cells were induced in vitro by the addition of anti-CD154 and anti-CD80 monoclonal antibodies (mAbs) to primary MLR (mixed lymphocyte reaction) consisting of BALB/c as responder and C3H as stimulator. Anergic cells were added to a newly formed MLR in assessing the regulatory capacity and antigen specificity of anergic cells. The ability of anergic cells to respond to antigen and/or exogenous recombinant mouse interleukin-2 (rmIL-2) was tested. For in vivo studies, anergic cells were intravenously injected into 3.0-Gy gamma-irradiated BALB/c mice immediately after heterotopic abdominal cardiac transplantation. To prolong allograft survival, recipient mice injected with anergic cells received rapamycin therapy [1 mg.day(-1).kg(-1)]. RESULTS: Anergic cells strongly suppressed the proliferation of naicaron;ve BALB/c splenocytes against the original (C3H) stimulator in a dose-dependent manner, but they failed to suppress the proliferation of naicaron;ve BALB/c splenocytes against the third-party (C57BL/6J) stimulator. The anergic state was reversed by both original (C3H) stimulator and additional exogenous IL-2. In in vivo studies, untreated irradiated BALB/c mice rejected C3H cardiac allografts with a mean survival time of (8.6 +/- 1.1) days, whereas those injected with the anergic cells rejected the allografts with a mean survival time of (11.8 +/- 1.9) days, which was slightly longer than that of the untreated mice. The protocol based on anergic cells injection plus rapamycin therapy could prolong allograft survival significantly [(29.6 +/- 4.4) days]. CONCLUSIONS: Anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro, and prolong cardiac allograft survival after adoptive transfer in the presence of rapamycin therapy. This procedure might be clinically useful for prolonging allograft survival if optimal protocols are developed. PMID- 15361292 TI - Transplantation tolerance mediated by regulatory T cells in mice. AB - BACKGROUND: With potent suppressive effect on responder T cells, CD(4)(+)CD(25)(+) regulatory T (Treg) cells have become the focus of attention only recently and they may play an important role in transplantation tolerance. However, the mechanism of action is not clear. This study was designed to assess the possibility of using CD(4)(+)CD(25)(+) Treg cells to induce transplantation tolerance and to investigate their mechanism of action. METHODS: CD(4)(+)CD(25)(+) Treg cells were isolated using magnetic cell separation techniques. Mixed lymphocyte reactions were used to assess the ability of Treg cells to suppress effector T cells. Before skin transplantation, various numbers of CD(4)(+)CD(25)(+) Treg cells, which have been induced using complex skin antigens from the donor, were injected into the host mice either intraperitoneally [0.5 x 10(5), 1 x 10(5), 2 x 10(5), 3 x 10(5), 4 x 10(5), or 5 x 10(5)] or by injection through the tail vein [5 x 10(3), 1 x 10(4), 2 x 10(4), 5 x 10(4), 1 x 10(5), 2 x 10(5)]. Skin grafts from two different donor types were used to assess whether the induced Treg cells were antigen-specific. The survival time of the allografts were observed. Single photon emission computed tomography was also used to determine the distribution of Treg cells before and after transplantation. RESULTS: Treg cells have suppressive effect on mixed lymphocyte reactions. Grafts survived longer in mice receiving CD(4)(+)CD(25)(+) Treg cell injections than in control mice. There was a significant difference between groups receiving intraperitoneal injection of either 2 x 10(5) or 3 x 10(5) CD(4)(+)CD(25)(+) Treg cells and the control group (P < 0.05, respectively). Better results were achieved when Treg cells were injected via the tail vein than when injected intraperitoneally. The transplantation tolerance induced by CD(4)(+)CD(25)(+) Treg cells was donor-specific. Analysis of the localization of Treg cells revealed that Treg cells mainly migrated from the liver to the allografts and the spleen. CONCLUSIONS: CD(4)(+)CD(25)(+)Treg cells can induce donor-specific transplantation tolerance. Cell-to-cell contact may be the primary mechanism by which Treg cells act on effector T cells. PMID- 15361293 TI - Effect of propentofylline on hypoxic-ischaemic brain damage in newborn rat. AB - BACKGROUND: Studies showed that propentofylline enhances the action of adenosine and protects hippocampal neuronal damage against transient global cerebral ischaemia. Our study was to investigate the effect of propentofylline on hypoxic ischaemic brain damage in neonatal rat. METHODS: Seven-day-old Wistar rats were subjected to unilateral common carotid artery ligation and hypoxia in oxygen 8 kPa for two hours at 37 degrees C. Propentofylline (10 mg/kg) was administered intraperitoneally one hour after hypoxia-ischaemia (treated group). Control group rats were received an equivalent volume of saline. The effects of propentofylline were assessed by observing the body mass gain, behavioural alteration and neurohistological changes. The rats were sacrificed at 72 hours after hypoxia ischaemia, and the brain sections were examined after haematoxylin and eosin staining. RESULTS: The propentofylline-treated rats had better body mass gain and better behavioural response than the paired saline-controls did. In the control group, the rats either lost body mass or had little mass gain after the insult, their average body mass gain was 97.3% at 24 h, 100.3% at 48 h, and 114.1% at 72 h of recovery. In propentofylline-treated group, there was a significant improvement of body mass gain at 24 h (100.2%, P < 0.05) and 48 h (110.3%, P < 0.01) of recovery; the percentage of rats that performed well on behavioural test was significantly higher from 48 h to 72 h of recovery (P < 0.05); the incidence of severe brain damage to the cerebral cortex and dentate gyrus was significantly reduced in propentofylline-treated rats (cortex, 93% - 70.8%, P < 0.01; dentate gyrus 95% - 66.7%, P < 0.01) as compared with control rats. CONCLUSIONS: Administration of propentofylline 1 hour after hypoxia-ischaemia significantly attenuates brain damage in both the cerebral cortex and dentate gyrus, and also improves the body mass gain as well as behavioural disturbance in 7-day-old rats. PMID- 15361295 TI - Effect of chemical treatment of silicon gel on tissue compatibility. AB - BACKGROUND: Silicon gel is unfavourable for cell attachment and growth. This study was to study if pretreating the surface of silicon gel with chemical agents affects the proliferation of epithelial cells. METHODS: Silicon gel was made and treated with either mixed acid solution (containing 232 g/dm(3) of H(2)SO(4) and 8 g/dm(3) of K(2)Cr(2)O(7)) or 300 cm(3)/dm(3) peroxide for 5, 10, and 15 minutes or 10, 15, and 20 minutes, respectively. The cultured corneal epithelial cells were seeded onto those silicon gels and kept for 13 days. Immunohistochemical investigations were then carried out for integrin (alpha 6 or beta 4) and actin. RESULTS: Growth of the epithelial cells in silicon gels treated with mixed acid solution for 10 minutes and 15 minutes was much significant than that in the untreated gels. After a 12-hour culture, a small number of corneal epithelial cells were proliferated on the surface of the silicon gels that had been treated with peroxide for 15 minutes. After a 3-day culture, those cells were further proliferated and fused together. The corneal epithelial cells did not grow well in the silicon gels treated with peroxide for 10 or 20 minutes. Immunostaining revealed the expression of actin and integrin alpha 6 or beta 4 on the silicon gels that were treated with mixed acid solution for 10 minutes or peroxide for 15 minutes. CONCLUSION: Silicon gels treated either with mixed acid solution for 10 or 15 minutes or with peroxide for 15 minutes improves cell proliferation. PMID- 15361294 TI - Dynamic changes in myocardial matrix metalloproteinase activity in mice with viral myocarditis. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are the major regulators of collagen degradation involved in the pathogenesis of several diseases of the heart. The purpose of this study was to investigate the dynamic changes in myocardial MMP activity in mice with viral myocarditis (VM), the relationship between MMP activity and both cardiac function and the quantity of myocardial collagen, and the role MMPs playing in the pathological lesions of VM. METHODS: Sixty-five six week-old male DBA/2 mice were divided into two groups. Mice in the infected group (n = 50) were inoculated intraperitoneally with 0.14 ml of Coxsackievirus B3 (CVB3, Nancy strain). Control mice (n = 15) were inoculated intraperitoneally with 0.14 ml of Eagle's medium. Eight infected mice and three control mice were sacrificed on each of days 3, 7, 10, 21 and 30 after inoculation. MMP activity was measured on an SDS-PAGE substrate gel embedded with type I gelatin (zymography). Echocardiographic studies were performed under anesthesia with 3% chloralhydrate administered intraperitoneally (0.01 ml/g - 0.015 ml/g). Cardiac systolic function indices, such as peak velocity of the aorta (Vp), flow velocity integral of the aorta (Vi), ejection fraction (EF), and fractional shortening (FS) were determined by echocardiography. Histological cross sections of the hearts were stained with hematoxylin-eosin and myocardial histopathological scores were determined under an optical microscope. The amount of myocardial collagen was measured by means of hydroxyproline quantification. RESULTS: In virus-infected mice, both MMP-2 and MMP-9 activities were significantly higher than in control mice, reaching a peak on day 10 (P < 0.01). On day 10, cardiac systolic function indices (EF, FS, Vp, and Vi) were all significantly lower compared both to other stages following viral inoculation and to the control group (P < 0.05). In the acute stage, the amount of myocardial collagen in mice with VM was not significantly different from normal control mice (P > 0.05). However, the amount of myocardial collagen in infected mice at the recovery stage (on days 21 and 30) was significantly greater than those of the control mice. MMP 2 and MMP-9 activities positively correlated with myocardial histopathological scores (r = 0.801, 0.821, P < 0.01) and negatively correlated with Vp (r = 0.649, -0.683, P < 0.01) and Vi (r = -0.711, -0.755, P < 0.01). However, Vp negatively correlated with myocardial histopathological scores (r = -0.756, P < 0.01). CONCLUSIONS: In mice with VM, the activities of myocardial MMP-2 and MMP-9 increase significantly during the acute stage, and the total quantity of myocardial collagen increases by the time of recovery. These changes are associated with myocardial interstition remodeling and cardiac dysfunction. MMP activity is an important reference marker for myocardial pathological lesions and can be used to evaluate the severity of myocardial interstitial damage and cardiac dysfunction. PMID- 15361296 TI - Detailed deletion mapping of loss of heterozygosity on 9p13-23 in laryngeal squamous cell carcinoma by microsatellite analysis. AB - BACKGROUND: This study was designed to investigate the hot spots of microsatellite loss of heterozygosity (LOH) on 9p13-23 in laryngeal squamous cell carcinoma and to find out the correlation between the incidence of microsatellite LOH and the clinicopathological parameters. METHODS: Tumor tissues were obtained from paraffin embedded sections with microdissection. Genomic DNA was extracted from tumor tissues and peripheral blood lymphocytes with the phenol-chloroform. Polymerase chain reaction (PCR) amplification and denaturing gel electrophoresis were carried out in a set of 42 squamous cell carcinoma (SCC) of larynx and corresponding peripheral blood lymphocytes using 13 highly polymorphic microsatellite markers on 9p13-23. The correlation was analyzed between microsatellite LOH at the high frequency on 9p13-23 and clinicopathological parameters in the patients with squamous cell carcinoma of larynx. RESULTS: Of the 42 laryngeal cancers, 41 (97.6%) showed LOH in at least one of the microsatellite markers tested on 9p13-23. The most frequently deleted marker was D9S162 in 17 of the 19 (89.5%) informative samples. The marker D9S171, which is located on 9p21, had LOH detected in 12 of the 15 informative cases (80.0%). LOH at the D9S1748 marker (closest to the p16 gene locus) was detected in 18 of the 36 informative cases (50.0%). Allelic deletion mapping revealed two minimal regions of LOH encompassing markers D9S161-D9S171 on 9p21 and IFNA-D9S162 on 9p22 23. Multiple LOH (> or = 4) on 9p21-23 was found more frequently in the patients under 60 years, with supraglottic SCC or cervical lymph node metastasis than those over 60 years, with glottic SCC or without cervical lymph node metastasis (P < 0.01 or 0.01, 0.05, respectively). On the contrary, there was no correlation between T stages or pathologic classification and the frequency of LOH on 9p21-23 in 42 SCC of Larynx. CONCLUSIONS: These findings imply the presence of at least two putative tumor suppressor genes on 9p13-23 in laryngeal SCC. Multiple genetic alterations are probably implicated in supraglottic SCC with cervical lymph node metastasis in younger patients. PMID- 15361298 TI - Expression and self-assembly of HCV structural proteins into virus-like particles and their immunogenicity. AB - BACKGROUND: The synthesis of virus-like particles (VLPs) provides an important tool to determine the structural requirements for viral particle assembly and virus-host interactions. Our purpose was to express simultaneously all three structural proteins of hepatitis C virus (HCV) in insect cells to investigate the proteins assembly into VLPs and the immunogenicity of these particles. METHODS: HCV gene sequences encoding the structural proteins C, E1, and E2 were amplified with PCR, and recombinant baculoviruses were constructed using recombinant DNA techniques. The expression of HCV structural proteins in insect cells was analyzed by immunofluorescence and SDS-PAGE. The interaction of expressed structural proteins was investigated by immunoprecipitation and immunoblotting. The VLPs in the insect cells were visualized by electron microscopy (EM). VLPs were then purified by sucrose gradient centrifugation and used to immunize BALB/c mice. Antibodies against HCV were tested for in mouse serum samples by an ELISA assay. RESULTS: The recombinant baculoviruses reBV/C and reBV/E1-E2 were constructed successfully. Insect cells co-infected with reBV/C and reBV/E1-E2 expressed HCV C, E1, and E2 proteins with the expected molecular weights of 20kD, 35kD, and 66kD, respectively. The results of immunoprecipitation and immunoblotting assays revealed the coimmunoprecipitation of C, E1, and E2 proteins, indicating association of the three structural proteins. Electron microscopy of insect cells co-infected with reBV/C and reBV/E1-E2 demonstrated spherical particles (40 to 60 nm in diameter) similar to the HCV virions from serum samples or hepatic tissue samples of HCV infected humans. The VLPs were partially purified. Antibodies to HCV were detectable in the serum of mice immunized with VLPs. CONCLUSION: HCV structural proteins simultaneously expressed in insect cells can interact with each other and assemble into HCV-like particles, which are shown to be immunogenic in mice. PMID- 15361297 TI - Expression of survivin mRNA in peritoneal lavage fluid from patients with gastric carcinoma. AB - BACKGROUND: Peritoneal dissemination is the most common pattern of metastasis in advanced gastric carcinoma with serosal invasion. In the present study, we reported the clinical relevance of a new diagnostic method involving RT-PCR, using survivin as the target gene, for the detection of free cancer cells in peritoneal washes. METHODS: Intraoperative peritoneal washes were obtained from 48 patients who underwent surgery for gastric cancer. RT-PCR analysis with primers specific for survivin and conventional cytological examinations were both performed. RESULTS: Survivin mRNA was not detected in any peritoneal wash samples from patients with benign disease, but was detected in 28 of 48 samples taken from patients with gastric cancer and in all metastatic nodules. Survivin expression in the peritoneal cavity significantly correlated with depth of cancer invasion, lymph node metastasis, and TNM stage. There were 92% of clinically evident peritoneal metastasis cases showed detectable survivin expression. The combination of survivin RT-PCR and cytological examination yielded positive results in 66.7% (32/48) of patients with gastric cancer, much higher than the results produced by cytological method alone. CONCLUSIONS: Survivin mRNA detected in peritoneal lavage fluid might indicate the presence of free cancer cells in the peritoneal cavity. The high sensitivity of the RT-PCR-based survivin assay suggests that survivin serves as a molecular marker for detecting peritoneal micrometastasis. Its ubiquitous expression in peritoneal cancer cells and metastatic nodules also suggests a promising future therapeutic strategy based on survivin inhibition for cases of gastric cancer involving peritoneal metastasis. PMID- 15361299 TI - Bone marrow stromal cell line co-transfected with IL-2 and IL-3 genes can accelerate restoration of T-cell immunity in allo-BMT mice. AB - BACKGROUND: After T-cell depleted allogeneic bone marrow transplantation, impaired immune reconstitution is a major cause of morbidity and mortality in the recipient. The purpose of this study was to observe the effects of the gene engineered bone marrow stromal cell line QXMSC1-IL-2 + IL-3 on the reconstitution of T-cell immunity in allo-BMT mice. METHODS: The bone marrow stromal cell line QXMSC1 was co-transfected with IL-2 and IL-3 genes using a Tet-on gene expression system. T lymphocyte subset counts per spleen were analyzed by flow cytometry. Lymphocyte proliferation response to ConA was examined to evaluate T-cell function. CDR3 spectratyping techniques were performed to evaluate TCR repertoire diversity at various time points post-transplantation. RESULTS: Gene engineered bone marrow stromal cell line QXMSC1-IL-2 + IL-3 could express IL-2 and IL-3 [1,300 ng.day(-1).10(-6) cells and 1100 ng.day(-1).10(-6) cells, respectively] under the control of doxycycline. QXMSC1-IL-2 + IL-3 in combination with allogeneic bone marrow could significantly increase the counts of CD(4)(+) and CD(8)(+) T cell, 1.72 and 1.27-fold respectively at week 3 compared with TCD-BMT group (P < 0.01); make CD(4)(+)/CD(8)(+) ratio return to normal level at week 4; enhance splenocytes mitotic response to ConA (P < 0.01), and accelerate restoration of TCR repertoire diversity in the lethally irradiated mice (P < 0.05). CONCLUSION: The gene transduced stromal cell line QXMSC1-IL-2 + IL-3 is able to accelerate T-cell immunity in allo-BMT mice. PMID- 15361300 TI - Microarray analysis of extracellular matrix genes expression in myocardium of mouse with Coxsackie virus B3 myocarditis. AB - BACKGROUND: Extracellular matrix (ECM) orchestrates cell behaviour including growth, death, apoptosis, adhesion, migration, and invasion by activating several signalling pathways. Certain components of ECM, such as integrins, may act as receptors or co-receptors of enterovirus. ECM-activated gene expressions in myocardium of viral heart disease including myocarditis and partial cardiomyopathy remain elusive. This study was to investigate the expression of ECM-activated genes in myocardium of mouse with viral myocarditis. METHODS: BALB/c mice were infected with Coxsackie virus B3 (CVB3) to establish an animal model of myocarditis. Uninfected mice were also prepared and served as controls. Specific mRNA expression pattern in myocarditic mouse heart was analysed by an in house cDNA microarray containing 8,192 genes. Overexpressed ECM genes were selected and subsequently confirmed by Northern blot analysis. RESULTS: Nine ECM genes were isolated, from the array of 8,192 genes, as overexpressed genes in hearts of myocarditic mice in comparison with controls. Subsequent Northern blot analysis confirmed that four of the nine genes were highly expressed. Expression of these four genes, Fin15, ILk, Lamr1 and ADAMTS-1, has not been reported previously to be induced by Coxsackie virus. CONCLUSION: CVB3-induced myocarditis is associated with gene expression profiles of certain ECM components. PMID- 15361301 TI - Association of E-cadherin and beta-catenin with metastasis in nasopharyngeal carcinoma. AB - BACKGROUND: This study was designed to detect methylation of E-cadherin gene promoter and gene mutation of beta-catenin in exon 3 and their expression of protein and mRNA in primary tumor and lymph node metastatic tumor of nasopharyngeal carcinoma (NPC), and investigate the mechanism of invasion and metastasis of neoplastic cells in NPC. METHODS: Fourty-two fresh biopsy samples were taken from untreated NPC patients at the Affiliated Hospital of Sun Yat-sen Medical College, Sun Yat-sen University, Guangzhou, China during the period of 1999-2002. Among them 21 were taken from primary tumors and the other 21 from lymph node metastatic tumors. The gene promoter methylation of E-cadherin was detected by methylation-specific PCR (MSP). The mutation in exon 3 of beta catenin was detected by direct sequencing analysis. RT-PCR, Western blot and immunohistochemical staining were used to detect the mRNA and protein expression patterns in both primary and metastatic tumors of NPC. RESULTS: Down-regulated expression of E-cadherin in metastatic tumor was compared with that in primary tumor. Reduced expression of E-cadherin was found to be correlated with lymph node metastatic tumor of NPC (P = 0.004); but there was no obvious correlation between primary and metastatic tumors in the expression of beta-catenin (P = 0.698). The mRNA expression level of E-cadherin in metastatic tumors decreased significantly compared with that in primary tumors. However, little change was observed in the mRNA level of beta-catenin in different tumor tissues. Only 4 samples (19.1%) displayed gene promoter methylation of E-cadherin in primary tumor and 10 samples (47.6%) showed methylated form of E-cadherin. The gene promoter methylation of E-cadherin was more common in metastatic tumor than in primary tumor of NPC (P = 0.024). Only 2 (4.76%) of the 42 samples showed mutations in exon 3 of beta-catenin at 41 (T41A, ACC-->GCC) and codon 47 (S47T, AGT-->ACT). The cytoplasmic and nuclear expression of beta-catenin in tumor was not found in any samples of NPC. CONCLUSIONS: The results suggest that the downregulation of E-cadherin results from the gene promoter aberrant methylation of E-cadherin and that the methylation of E-cadherin plays an important role in invasion and metastasis of tumor cells in NPC. However, beta-catenin mutation is an infrequent event in NPC, and beta-catenin is not a critical factor influencing the invasion and metastasis of tumor cells in NPC. PMID- 15361302 TI - Septation and shortening of outflow tract in embryonic mouse heart involve changes in cardiomyocyte phenotype and alpha-SMA positive cells in the endocardium. AB - BACKGROUND: Studies on human, rat and chicken embryos have demonstrated that during the period of outflow tract septation, retraction of the distal myocardial margin of the outflow tract from the junction with aortic sac to the level of semilunar valves leads to the shortening of the myocardial tract. However, the mechanism is not clear. So we investigated the mechanism of outflow tract shortening and remodeling and the spatio-temporal distribution pattern of alpha SMA positive cells in the outflow tract cushion during septation of the outflow tract in the embryonic mouse heart. METHODS: Serial sections of mouse embryos from embryonic day 9 (ED 9) to embryonic day 16 (ED 16) were stained with monoclonal antibodies against alpha-SCA, alpha-SMA, or desmin, while apoptosis was assessed using the terminal deoxyribonucleotidy transferase-mediated dUTP digoxigenin nick-end labeling (TUNEL) assay. RESULTS: Between ED 11 and ED 12, the cardiomyocytes in the distal portion of the outflow tract were observed losing their myocardial phenotype without going into apoptosis, suggesting that trans-differentiation of cardiomyocytes into the cell components of the free walls of the intrapericardial ascending aorta and pulmonary trunk. The accumulation of alpha-SMA positive cells in the cardiac jelly began on ED 10 and participated in the ridge fusion and septation of the outflow tract. Fusion of the distal ridges resulted in the formation of the facing walls of the intrapericardial ascending aorta and pulmonary trunk. Fusion of the proximal ridges was accompanied by the accumulation of alpha-SMA positive cells into a characteristic central whorl, in which cell apoptosis could be observed. Subsequent myocardialization resulted in the formation of the partition between the subaortic and subpulmonary vestibules. CONCLUSIONS: The shortening of the embryonic heart outflow tract in mice may result not from apoptosis, but from the trans-differentiation of cells with cardiomyocyte phenotype in the distal portion of the outflow tract into the cell components of the free walls of the intrapericardial ascending aorta and pulmonary trunk. The primary roles of alpha SMA positive cells in the septation and remodeling of the outflow tract may assure proper fusion of the outflow ridges and form the facing walls of the intrapericardial ascending aorta and pulmonary trunk. PMID- 15361303 TI - Study on the pathological basis of classification of spleen deficiency in chronic gastritis. AB - BACKGROUND: Spleen in Traditional Chinese Medicine (TCM) is not actually the spleen in the anatomic sense designated in western medicine because its functions basically belong to the physiological category of digestive system in modern medicine, and it represents a macroscopic concept of digestion, absorption and nutrition metabolism. Spleen deficiency syndrome refers to the clinical phenomena such as hypofunction of digestion, absorption and nutrition metabolism. By integrating TCM with modern medicine, this paper is intended to explore the pathological basis of classification of spleen deficiency in chronic gastritis. METHOD: By means of optical microscope, scanning electron microscope (SEM), transmission electron microscope (TEM) and histochemical staining, we conducted histopathological and subcellular ultrastructural (nuclei and mitochondrial) analysis of gastric mucosa of 188 patients of spleen deficiency, and that of 42 voluntary blood donors without clinical symptoms. RESULTS: The gastric mucosa of patients with spleen Qi deficiency (SQD) and spleen yang deficiency (SyangD) could either be affected by organic lesion (type G-occurring on the basis of chronic superficial gastritis (CSG), chronic atrophic gastritis (CAG)) or unaffected (type F-chiefly belonging to functional indigestion); spleen yin deficiency (SyinD) and spleen deficiency with Qi stagnation (SDQS) both occurred on the basis of CSG and CAG; and the degree of mucosa inflammatory cells infiltration, the degree of decrease in glands propria, and the incidence of IMIIb in CSG and CAG were more serious than those of G-SQD and G-SyangD, P < 0.05 - 0.01. CONCLUSION: Spleen deficiency syndrome is likely to occur on the basis of organic lesion of gastric mucosa (disease with symptoms of both CSG or CAG and spleen deficiency symptoms), as well as on the basis of inorganic lesion of gastric mucosa (nondisease with symptoms, which is, despite spleen deficiency symptoms, there is no CSG or CAG). Besides, the clinical phenomenon of disease without symptoms (despite CSG or CAG, there is no spleen deficiency symptoms) occurres because of such factors as genetic diathesis and compensation. The lesion degree of CSG or CAG and the incidence of IMIIb of SyinD and SDQS are more serious than those CSG and CAG of G-SQD and G-SyangD. PMID- 15361304 TI - PPAR-gamma ligands inhibit the expression of inflammatory cytokines and attenuate autoimmune myocarditis. PMID- 15361305 TI - The relationship between gap junctional remodeling and human atrial fibrillation. PMID- 15361306 TI - Characteristics of mRNA levels of hepatic key enzymes in cholesterol metabolism of genetically gallstone-susceptible mice. PMID- 15361307 TI - Multiple endocrine neoplasia type 2B caused by a single point mutation in RET proto-oncogene in a Chinese patient. PMID- 15361308 TI - Computer assisted reconstruction of three-dimensional canal model of femur and design for custom-made stem. PMID- 15361309 TI - Growth and development of term infants fed with milk with long-chain polyunsaturated fatty acid supplementation. PMID- 15361310 TI - Activity of dihydroartemisinin against Leishmania donovani both in vitro and vivo. PMID- 15361311 TI - Suboccipital bony decompression combined with removal of the dural band as treatment for Chiari I malformation. PMID- 15361312 TI - Death of a SARS case from secondary aspergillus infection. PMID- 15361313 TI - Rituximab (MabThera) for aggressive non-Hodgkin's lymphoma: systematic review and economic evaluation. AB - OBJECTIVES: To determine the clinical and cost-effectiveness of adding rituximab to the CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone) chemotherapy regime for adult patients with diffuse large B-cell lymphoma (DLBCL). DATA SOURCES: Electronic bibliographic database. REVIEW METHODS: Comparative studies were selected for review if they addressed the clinical or cost-effectiveness of adding rituximab to CHOP in people aged at least 18 years with DLBCL. The internal validity of the study was assessed through the use of the validated Jadad scoring system. Data were abstracted into standardised data extraction forms. Costs were estimated through resource use data taken from the published trial and the unpublished sponsor submission. Unit costs were taken from published sources, where available. An economic evaluation was undertaken to evaluate the cost-effectiveness of R-CHOP compared with CHOP alone for patients with DLBCL using data sources and methodology similar to the manufacturer's submission. RESULTS: In the systematic review of effectiveness, one randomised controlled trial was identified. The study was, in most respects, methodologically rigorous and well conducted and the statistical evidence favoured the addition of rituximab to CHOP. The total cost of rituximab with CHOP (R-CHOP) and CHOP alone estimated from the model developed by ScHARR was 14,456 pounds and 5773 pounds, respectively, for patients aged 60 years and over, and 15,181 pounds and 7311 pounds for patients aged less than 60 years over a 15-year time horizon. The ScHARR model estimated that the addition of rituximab to CHOP generated an additional 0.82 QALY at an extra cost of 8683 pounds compared with CHOP alone therapy over a 15-year time horizon, a cost/quality-adjusted life-year (QALY) ratio of 10,596 pounds for patients aged 60 years or more. For patients aged under 60 years, 1.05 QALY were generated at an additional cost of 7870 pounds, a cost/QALY ratio of 7533 pounds. Assuming that the societal value of a QALY was 30,000 pounds then R-CHOP is cost-effective compared with CHOP in the treatment of DLBCL. CONCLUSIONS: In the short term, the addition of rituximab to the CHOP regimen increased the likelihood of a complete-response by 20% without a significant rise in the risk of a serious adverse event in people aged 60 years or older. Over a 2-year follow-up period, the intervention reduced the risk of death, progression or relapse by 45% and reduced the risk of death by 47% in this population. There is no direct evidence for the clinical effectiveness of adding rituximab to CHOP in the treatment of DLBCL in those aged 18-59 years, although data from phase I and II trials confirm its safety and efficacy in a preclinical setting. The cost-effectiveness modelling presented here has shown that rituximab in combination with CHOP chemotherapy regimen is likely to be considered a cost effective treatment for DLBCL when compared with the current standard treatment, CHOP chemotherapy only. Analysis of quality of life (QoL) in the area of NHL is limited and only one cost-utility analysis for the treatment of CHOP in NHL was identified. Both the SCHARR and the manufacturer's models utilised QoL utility scores from an unpublished data source. Further research within this area would help to improve the robustness of QoL utility analysis within DLBCL and also NHL as a whole. Further clinical trials might also establish whether R-CHOP may replace peripheral blood stem cell transplant in high-risk patients and whether the doses of chemotherapy in the elderly may be reduced if rituximab is added to less intensive regimens. PMID- 15361314 TI - Review of guidelines for good practice in decision-analytic modelling in health technology assessment. AB - OBJECTIVES: To identify existing guidelines and develop a synthesised guideline plus accompanying checklist. In addition to provide guidance on key theoretical, methodological and practical issues and consider the implications of this research for what might be expected of future decision-analytic models. DATA SOURCES: Electronic databases. REVIEW METHODS: A systematic review of existing good practice guidelines was undertaken to identify and summarise guidelines currently available for assessing the quality of decision-analytic models that have been undertaken for health technology assessment. A synthesised good practice guidance and accompanying checklist was developed. Two specific methods areas in decision modelling were considered. The first method's topic is the identification of parameter estimates from published literature. Parameter searches were developed and piloted using a case-study model. The second topic relates to bias in parameter estimates; that is, how to adjust estimates of treatment effect from observational studies where there are risks of selection bias. A systematic literature review was conducted to identify those studies looking at quantification of bias in parameter estimates and the implication of this bias. RESULTS: Fifteen studies met the inclusion criteria and were reviewed and consolidated into a single set of brief statements of good practice. From this, a checklist was developed and applied to three independent decision analytic models. Although the checklist provided excellent guidance on some key issues for model evaluation, it was too general to pick up on the specific nuances of each model. The searches that were developed helped to identify important data for inclusion in the model. However, the quality of life searches proved to be problematic: the published search filters did not focus on those measures specific to cost-effectiveness analysis and although the strategies developed as part of this project were more successful few data were found. Of the 11 studies meeting the criteria on the effect of selection bias, five concluded that a non-randomised trial design is associated with bias and six studies found 'similar' estimates of treatment effects from observational studies or non-randomised clinical trials and randomised controlled trials (RCTs). One purpose of developing the synthesised guideline and checklist was to provide a framework for critical appraisal by the various parties involved in the health technology assessment process. First, the guideline and checklist can be used by groups that are reviewing other analysts' models and, secondly, the guideline and checklist could be used by the various analysts as they develop their models (to use it as a check on how they are developing and reporting their analyses). The Expert Advisory Group (EAG) that was convened to discuss the potential role of the guidance and checklist felt that, in general, the guidance and checklist would be a useful tool, although the checklist is not meant to be used exclusively to determine a model's quality, and so should not be used as a substitute for critical appraisal. CONCLUSIONS: The review of current guidelines showed that although authors may provide a consistent message regarding some aspects of modelling, in other areas conflicting attributes are presented in different guidelines. In general, the checklist appears to perform well, in terms of identifying those aspects of the model that should be of particular concern to the reader. The checklist cannot, however, provide answers to the appropriateness of the model structure and structural assumptions, as these may be seen as a general problem with generic checklists and do not reflect any shortcoming with the synthesised guidance and checklist developed here. The assessment of the checklist, as well as feedback from the EAG, indicated the importance of its use in conjunction with a more general checklist or guidelines on economic evaluation. Further methods research into the following areas would be valuable: the quantification of selection bias in non-controlled studies and in controlled observational studies; the level of bias in the different non-RCT study designs; a comparison of results from RCTs with those from other non-randomised studies; assessment of the strengths and weaknesses of alternative ways to adjust for bias in a decision model; and how to prioritise searching for parameter estimates. PMID- 15361315 TI - Coronary artery stents: a rapid systematic review and economic evaluation. AB - OBJECTIVES: To assess the effectiveness and cost-effectiveness of the use of coronary artery stents in patients with coronary heart disease (CHD). DATA SOURCES: Electronic databases. REVIEW METHODS: The review was conducted following accepted guidelines for conducting systematic reviews. Randomised controlled trials that include comparisons of percutaneous transluminal coronary angioplasty (PTCA) versus PTCA with stent, stent versus coronary artery bypass graft (CABG), and drug-eluting stents (DES) versus non-DES in patients with CAD in native or graft vessels and those with stable angina or acute coronary syndrome (ACS) and unstable angina were also included. Data on the following outcome measures were included in the review: combined event rate or event-free survival, death, acute myocardial infarction, target vessel revascularisation, repeat treatment (PTCA, stent or CABG) and binary restenosis. An economic model was developed based on extrapolation of trends in mortality and revascularisation from clinical trials data to a 5-year time horizon. RESULTS: The inclusion criteria were fulfilled by 50 studies comparing the use of stents with PTCA, six comparing stents with CABG and 12 comparing DES eluting stents with non-DES. No studies were identified that compared DES with PTCA or DES with CABG. Existing quality of life data suggest that revascularisation procedures reduce the patient's quality of life for a short period only. Stents were found to be more effective than PTCA in preventing adverse events and revascularisations. In multiple-vessel disease there was no evidence of a difference in mortality (at 1 year) between patients treated surgically and those receiving a stent. Patients treated surgically required fewer revascularisations. There is no evidence of a difference in mortality between patients receiving DES and those treated with bare metal stents at 1 year. A reduction in event rate at 9 and 12 months was found in patients treated with DES. This event rate is primarily made up of increased revascularisation rates in patients treated with bare metal stents. Two-year outcome data from one study indicate that this benefit of DES continues over the longer term. The economic model proved sufficient to indicate long-term trends in cost effectiveness. CABG was found initially to be more expensive than bare metal stenting in multivessel disease and may have higher immediate risks, but over time the cost differential is reduced and long-term outcomes favour CABG over stenting. A similar situation was found for DES versus CABG in multiple-vessel disease. However, DES may not generally be considered a cost-effective alternative to bare metal stenting in single-vessel disease by policy makers as substantially higher costs are involved with a very small outcome benefit. CONCLUSIONS: DES might be considered cost-effective if the additional cost (compared with ordinary stents) was substantially reduced, the outcome benefits from the use of DES were much improved, and/or its use were targeted on the subgroups of patients with the highest risks of requiring reintervention. Long term clinical studies are needed that focus on significant outcomes such as mortality. Further research should consider: the differences among plain stents; head-to-head comparisons within DES, CABG compared with DES; and the evaluation of newer non-DES against DES. Evaluation of the effects of revascularisation procedures and especially repeat revascularisation procedures on the patient's quality of life would also be useful, as would the development and testing of risk assessment tools to identify patients likely to need further revascularisations. PMID- 15361316 TI - Evaluation of abnormal uterine bleeding: comparison of three outpatient procedures within cohorts defined by age and menopausal status. AB - OBJECTIVES: To compare three outpatient methods of endometrial evaluation in terms of performance, patient acceptability and cost-effectiveness. DESIGN: Pragmatic unblinded trial randomised separately within three groups determined by risk of endometrial cancer. SETTING: The gynaecology outpatient clinic of a large city hospital in Edinburgh, Scotland. PARTICIPANTS: Women referred for investigation and management of abnormal bleeding between January 1999 and May 2001. INTERVENTIONS: Investigations were: blind biopsy alone, hysteroscopy with biopsy, ultrasound evaluation including transvaginal ultrasound, and, in the low risk group, the option of no investigation. Within this design, two devices for obtaining endometrial biopsy were compared, the Pipelle sampler and the Tao brush. MAIN OUTCOME MEASURES: Successful (informative) completion of the investigation, acceptability of the investigation method to women, women's satisfaction with clinic care in the short term and at 10 months and 2 years of follow-up, and cost-effectiveness to the end of investigation. RESULTS: Minor adverse events (e.g. shock, patient distress) did not occur for ultrasound, but occurred in 16% and 10% of women for hysteroscopy and biopsy procedures respectively. Pipelle biopsy provided an acceptable endometrial sample for 79% of moderate-risk women, but only 43% of high-risk women. The Tao brush gave similar performance in moderate-risk women (77%), but was more successful than the Pipelle sampler in postmenopausal (high-risk) women (72%). There were significantly more successful visualizations for ultrasound than for hysteroscopy in both the low-risk and the moderate-risk group, and a similar but non significant trend in the high-risk group. Ultrasound was significantly better than hysteroscopy at detecting fibroids, but hysteroscopy significantly better for polyps. At the 10-month follow-up, high-risk women who had been investigated by hysteroscopy (with biopsy) had the most positive views of their clinic experience, but this effect had largely disappeared by 24 months. In the moderate risk group, the subgroup randomised to biopsy alone gave the most negative responses about their clinic experience and health now. Women wishing they had more investigation comprised 22% of moderate-risk women and 38% of low-risk women, but only 14% of postmenopausal women. At follow-up the moderate-risk women (with menstrual bleeding problems), compared with postmenopausal women, had much worse ratings for clinic experience and health now. Resource use tended to be higher in the moderate- and low-risk women. There was minimal difference in cost effectiveness between investigation options in the high-risk group, with the option involving hysteroscopy being marginally better than ultrasound. The most cost-effective investigation in the moderate-risk group was biopsy alone and in the low-risk group ultrasound. CONCLUSIONS: Decision-making about investigation would be clarified if postmenopausal women were studied separately from premenopausal women with menstrual bleeding problems. For postmenopausal women exclusion of cancer is a main objective, so once investigation has been completed discharge follows, but in the woman with abnormal menstrual bleeding, even if serious pathology is excluded, the original presenting symptoms require management. About 60% of premenopausal women with abnormal bleeding reported that their symptoms were not 'much improved' at 10 months. Research is needed to understand this phenomenon, and to explore ways to integrate patient factors into optimising evaluation and treatment. The significance of benign pathologies in this group also requires clarification. Given the relatively small differences observed in cost-effectiveness, there is justification for allowing other issues (such as clinician preferences and women's perspectives) to influence decisions as to the investigation method. There is scope to make better use of patient factors to inform decisions as to the most efficient and acceptable method of investigation for an individual woman. Additional analyses, using data available as a result of this study, will contribute to this agenda. PMID- 15361317 TI - Intraoral neuropathy. AB - Chronic nonodontogenic toothache has been reported in the literature since the 1700s. This problem has followed a similar scenario since those first reports. The patient typically is misdiagnosed and then subjected to multiple unnecessary procedures, ultimately resulting in tooth extractions because of dentists and physicians being unaware of the existence of atypical odontalgia and other types of intraoral neuropathic pain that are treatable without sacrificing the teeth. This paper reviews the medications and procedures used to treat nonodontogenic toothache. PMID- 15361318 TI - Chronic cluster headache: new and emerging treatment options. AB - Cluster headache (CH) is a primary headache syndrome characterized by short lasting unilateral head pain attacks accompanied by ipsilateral oculofacial autonomic phenomena. Approximately 20% of CH patients have the chronic form and need continuous medical care. In the chronic form, attacks continue unabated for years, often on a daily basis, resulting in severe debilitation. It is a common experience that drug treatments are able to control or prevent the attacks in approximately 80% of chronic CH patients. In the remaining 20% of chronic cases, drugs are ineffective. Until recently, the etiology of CH was poorly understood and this hampered the development of new therapies. However, we have now gained a much improved understanding of the peripheral and central mechanisms giving rise to the pain in CH and this has inspired the development of new treatment approaches, which, although still in the initial phases of validation, appear to be very promising. Among these, the novel approach based on hypothalamic deep brain stimulation is one of the most promising. PMID- 15361319 TI - The relationship of temporomandibular disorders and fibromyalgia: implications for diagnosis and treatment. AB - Although most cases of temporomandibular disorders (TMD) are mild and self limiting, approximately 10% of patients develop severe disorders associated with chronic pain. It has been found that the widespread pain, depression, and sleep disorders associated with fibromyalgia (FM) may play a significant role in the chronicity of patients with TMD. This paper reviews the characteristics and relationship between TMD and FM and discusses how the similar mechanisms and diagnostic and treatment strategies for both disorders suggest that there is a close relationship between them. PMID- 15361320 TI - Fibromyalgia: the gastrointestinal link. AB - Patients with fibromyalgia (FM) frequently have gastrointestinal symptoms and signs. This article critically reviews the available literature and concludes the following: evidence that inflammatory bowel disease is associated with FM is contradictory, but should be looked for in patients taking concomitant steroids; patients diagnosed with celiac disease often have a history of FM or irritable bowel syndrome (IBS) that may or may not be present; reflux, nonulcer dyspepsia, and noncardiac chest pain are common in FM patients; medications used to manage pain, inflammation, and gastrointestinal complaints confound the management of FM; and IBS affects smooth muscles and the parasympathetic nervous system, while FM patients have complaints of striated muscles and dysfunction of the sympathetic nervous system. Of those patients with FM, 30% to 70% have concurrent IBS. Small intestinal bacterial overgrowth is associated with hyperalgesia and IBS-like complaints, is common in FM, and responds transiently to antimicrobial therapy. PMID- 15361321 TI - History of fibromyalgia: past to present. AB - Fibromyalgia syndrome (FMS) is now a recognized clinical entity causing chronic and disabling pain. For several centuries, muscle pains have been known as rheumatism and then as muscular rheumatism. The term fibrositis was coined by Gowers in 1904 and was not changed to fibromyalgia until 1976. Smythe laid the foundation of modern FMS in 1972 by describing widespread pain and tender points. The first sleep electroencephalogram study was performed in 1975. The first controlled clinical study with validation of known symptoms and tender points was published in 1981. This same study also proposed the first data-based criteria. The important concept that FMS and other similar conditions are interconnected was proposed in 1984. The first American College of Rheumatology criteria were published in 1990 and neurohormonal mechanisms with central sensitization were developed in the 1990s. Serotonergic/norepinephric drugs were first shown to be effective in 1986. PMID- 15361322 TI - Fibromyalgia: present to future. AB - There has been a dramatic increase in our understanding of fibromyalgia throughout the past 14 years since the publication of the 1990 American College of Rheumatology classification criteria. Before 1990, and for most of the 20th century, fibromyalgia was considered to be predominantly a muscle disorder; now the critical abnormality is described as "central sensitization." However, central sensitization has to have an initial genesis and nociceptive stimuli from painful foci in muscle are increasingly recognized as being relevant to the development of fibromyalgia. Clinicians also recognize an association between the initiation of fibromyalgia and chronic psychologic stressors and inflammatory disorders. It has been more difficult to understand how two such apparently diverse events could affect central pain physiology. However, some clues are emerging from the role of diverse stimuli in activating glial cells and the role of disordered cytokine networks. Some predictions about future developments in fibromyalgia are ventured based on the current state of knowledge. PMID- 15361323 TI - Fibromyalgia as a sympathetically maintained pain syndrome. AB - Abnormal activity of the sympathetic nervous system may be involved in the pathogenesis of chronic pain syndromes. This article reviews the animal studies of sympathetically induced pain behavior, the controversy of sympathetically maintained pain in clinical practice, and the dysautonomic nature of fibromyalgia (FM). FM has neuropathic pain features (stimuli-independent pain state accompanied by allodynia and paresthesias). The proposal of FM as a sympathetically maintained pain syndrome is based on the controlled studies showing that patients with FM display signs of relentless sympathetic hyperactivity and that the pain is submissive to sympathetic blockade and is rekindled by norepinephrine injections. Dysautonomia also may explain the multisystem features of FM. PMID- 15361325 TI - Role of anesthesiologic blockade in headache management. AB - Local anesthetic blocks have been used in a variety of ways in the management of headache. Some practitioners use blocks of the supraorbital and greater occipital nerves as a form of treatment. Although the effect of each block is limited in duration, a series of injections appears to provide sustained relief. However, no controlled studies have vindicated blocks as a tenable intervention. On the other hand, blocks can be used to diagnose cervical sources of headaches. When performed under controlled conditions, such blocks can identify cervical sources of pain in a substantial proportion of patients with headache after whiplash. Positive blocks also predict good outcome from radiofrequency neurotomy of the nerve blocked. PMID- 15361324 TI - Post-triptan era for the treatment of acute migraine. AB - There now is one realized and several attractive targets for the treatment of acute attacks of migraine that will follow and augment the use of serotonin 5 HT1B/1D receptor agonists, the triptans. Calcitonin gene-related peptide (CGRP) receptor blockade recently has been shown to be an effective acute antimigraine strategy; therefore, blockade of CGRP release by inhibition of trigeminal nerves would seem a logical approach. A number of targets are reviewed in this article including serotonin 5-HT1F and 5-HT1D receptors, adenosine A1 receptors, nociceptin, vanilloid TRPV1 receptors, and anandamide CB1 receptors. Development of one or more such compound offers the exciting prospect of new non vasoconstrictor treatments for migraine and cluster headache. PMID- 15361326 TI - Role of neuroimaging in our understanding of the pathogenesis of primary headaches. AB - This article reviews new advances in neuroimaging of primary headaches. Imaging of the brain is reaching a new stage of maturity as the basic neural systems that participate in the pathogenesis of headaches are identified. Given the rapid advances in functional neuroimaging, it is no surprise that recent studies have supported the neurovascular theory of migraine and cluster headache. It is clear that functional neuroimaging will continue to be of paramount importance and ultimately may serve as the bridge between molecular and clinical domains in the field of headache research. PMID- 15361328 TI - [Schrodinger's query]. PMID- 15361327 TI - Cortical spreading depression: its role in migraine pathogenesis and possible therapeutic intervention strategies. AB - Cortical spreading depression (CSD) is a well-characterized phenomenon in experimental animals. Recent data show that CSD actually can occur in the injured human brain and compelling evidence is accumulating to support the concept that CSD is responsible for migraine aura. The aim of this review is to highlight recent key advances regarding our understanding of CSD in animal and human studies and its relevance to the pathophysiology of migraine and its potential treatment options. PMID- 15361329 TI - [Language]. PMID- 15361330 TI - [Cell therapy in leg ischemia]. PMID- 15361331 TI - [Identification of a new leprosy susceptibility gene with positional cloning]. PMID- 15361332 TI - [Do pancreatic stem cells exist?]. PMID- 15361333 TI - [ATG genes and macroautophagy]. PMID- 15361334 TI - [Quantum dots in biology: recent progress]. PMID- 15361335 TI - [Curing mosquitoes to control malaria?]. PMID- 15361336 TI - [Pyloric stenosis and neuronal nitric oxide synthase]. PMID- 15361337 TI - [A novel antifreeze protein in fish in the North Atlantic]. PMID- 15361338 TI - [Control of maintenance of neural stem cells in mammals]. PMID- 15361339 TI - [Predicting response to Iressa: unexpected mechanism]. PMID- 15361340 TI - [Is the extreme antigenic variability of P. falciparum facilitated by the host?]. PMID- 15361341 TI - [Dog as a mammalian genetic model]. AB - Up to recently, studies on dog genetics were rather scare notwithstanding the enormous potential that the canine model can offer in the study of the genotype/phenotype relationship and the analysis of the causes of many genetic diseases, with simple or complex inheritance, that affect dogs but also the human population. This potentiality is essentially due to the natural history of dogs whose domestication from wolves dated back 15,000 years, at least. All modern dogs originated from a limited number of female wolves from Eastern Asia. By applying a combination of selections and strong inbreeding practices, humans have created over 350 breeds, each of them corresponding to a genetic isolate and altogether offering a unique panel of polymorphism never encountered in any other mammals. In this review we summarized what makes dogs an unavoidable model. Contrary to the classical models like the two yeasts, nematode, fish, fly, mouse, or rat mainly used to understand the function of genes, dog with the creation across the centuries of numerous breeds offers a unique opportunity to study the role of their alleles. We report recent data on the construction of genomic maps and on the sequencing program of the dog genome launched by the National Institute of Health (NIH). To take fully advantage of the canine model, we advocate for the systematic construction of a rich canine single nucleotide polymorphisms (SNP) ressource to perform linkage desiquilibrium studies of normal or pathological traits as well as to get insight into the genetic diversity of the canine species. PMID- 15361342 TI - [Regulation of the expression of repeated sequences and RNA interference]. AB - Living organisms have to fight against the invasion of many parasites. Among them are viruses and transposable elements that are able to integrate in the genome of their host. After integration, they can replicate and propagate. The defence mechanisms against these invaders are still largely unknown but are widely studied in plants as well as in fungi and animals, particularly Caenorhabditis elegans and Drosophila melanogaster. The compilation of recent data allows us to draw a general scheme for these mechanisms. In particular, it seems that the propagation of viruses, transposable elements and repeated sequences is controlled by mechanisms repressing the expression of these sequences at both transcriptional and post-transcriptional levels. Post-transcriptional regulation of repeated sequences and transposable elements is related to RNA interference (RNAi), an evolutionary conserved silencing process induced by the presence of double-stranded RNAs (dsRNAs). Many genes and molecular intermediates have now been identified, revealing the different steps of the mechanism underlying this process. Finally, different results suggest that these post-transcriptional silencing processes are involved in the development of organisms. DICER is a nuclease responsible for the processing of dsRNA into short RNA molecules (siRNAs) considered as the interfering agent. siRNAs interact with the transcript of the targeted gene and this interaction induces degradation of the transcript. DICER is also involved in the processing of small temporal RNAs (stRNA) involved in the timing of development. stRNAs have the same structure as siRNAs. They regulate their target genes by interacting with elements present in their 3'UTR and blocking translation. RNAi appears to be an universal regulatory mechanism that was still unknown a few years ago. It is now largely used in large scale inactivation of genes to determine their function, and some recent studies indicate that it might be used in human therapy. PMID- 15361343 TI - [RAMPs and G protein coupled receptors]. AB - RAMPs (receptor activity-modifying proteins) were discovered in 1998 as accessory proteins needed to the functionnal activity of CGRP (calcitonin gene-related peptide) receptors. Three RAMPs generated by three different genes are known in human, rat and mice. The coding sequences of such genes are described, but as yet, regulation sequences are unknown. RAMPs interact with GPCR (G protein coupled receptors) of class II. In the case of the calcitonin/CGRP peptide family, RAMPs determine the functionnal specificity of the receptor, glycosylate and translocate the receptor to the cell surface. CGRP receptors are observed in presence of the RAMP1/calcitonin receptor-like receptor (CRLR), but the association of RAMP2 or RAMP3 with CRLR generates an adrenomedullin receptor. The calcitonin receptor (CTR) is translocated alone to the cell surface, but interactions of RAMPs with CTR forms amylin receptors. If RAMPs can interact with glucagon, parathyroid hormone and VIP/PACAP (vasoactive intestinal peptide/pituitary adenylate cyclase activating polypeptide (VPACR1)) receptors, the functionnal specificity of these receptors remains unaltered. However, the complex VPACR1/RAMP2 enhances specifically the phosphoinoside signaling pathway. PMID- 15361344 TI - [Mitochondria and reproduction]. AB - Mitochondria play a primary role in cellular energetic metabolism. They possess their own DNA, which is exclusively maternally transmitted. The relatively recent idea that mitochondria may be directly involved in human reproduction is arousing increasing interest in the scientific and medical community. It has been shown that the functional status of mitochondria contributes to the quality of oocytes and spermatozoa, and plays a part in the process of fertilisation and embryo development. Moreover, new techniques, such as ooplasm transfer, compromise the uniquely maternal inheritance of mitochondrial DNA, raising important ethical questions. This review discusses recent information about mitochondria in the field of human fertility and reproduction. PMID- 15361345 TI - [For a better understanding of the mechanisms involved in vertical transmission of HIV]. AB - Maternal-infant transmission of human immunodeficiency virus-1 (HIV) is the primary cause of this retrovirus infection in neonates. The mechanisms of vertical transmission of HIV, in particular in utero transmission, remain poorly defined. Trophoblastic cells from the placenta are thought to be a target of HIV infection and/or may be utilized by the virus to be transported across the placental barrier by a process known as transcytosis. The vertical transmission of HIV (via infection or transcytosis) may be either favoured or inhibited by factors related to both the viral phenotype and the cellular environment. PMID- 15361346 TI - [The supply of breast/ovarian cancer genetic susceptibility tests in France]. AB - One example of the recent advances of scientific research on the human genome is the identification of two susceptibility genes to breast/ovarian cancer, BRCA1 and BRCA2, making possible the introduction in medical practices of genetic testing to detect patients with an increased risk of developing such cancers. In this context of diffusion, two surveys were carried out to appraise the activity profiles in 1998 and in 2001 of all the different participants in those new medical practices in France, physicians in charge of genetic counselling, medical centres where consultations take place and laboratories. Results show that over the period 1998-2001, few changes occurred, mainly the reduction of the average waiting time to get the result of a genetic test, the increase in the annual number of BRCA2 families identified to a level similar to the one of BRCA1 and the automation of the biological analyses without noting a considerable increase in the annual output of laboratories till 2001 however. This surprising moderate evolution must be connected to the existence of some particular external factors making the framework of the development of these new medical and biological practices and their future really uncertain. The diffusion of BRCA1/2 genetic testing has been carried out facing the traditional difficulties of any innovating activities, but also the uncertainties related to intellectual property rights on genes and the reimbursement of genetic counselling and biological testing. These uncertainties have certainly restrained the pace of change as many actors in this field have opted for a wait and see strategy bearing in mind the possible future constraints imposed to their future activity, especially if European patents on the BRCA1/2 genes are finally granted by the European patent office (EPO). PMID- 15361347 TI - [Kallmann De Morsier syndrome: FGF-signaling insufficiency?]. AB - Kallmann syndrome (KAL) associates hypogonadotropic hypogonadism and anosmia, i.e. a deficiency of the sense of smell. Anosmia is related to the absence or the hypoplasia of the olfactory bulbs. Hypogonadism is due to GnRH deficiency, and is likely to result from the failed embryonic migration of GnRH-synthesizing neurons. These cells normally migrate from the olfactory epithelium to the forebrain along the olfactory nerve pathway. Kallmann syndrome is genetically heterogeneous. The gene responsible for the X-chromosome linked form of the disease, KAL-1, has been identified in 1991. KAL1 encodes a ~95 kDa glycoprotein of unknown function, which is present locally in various extracellular matrices during the period of organogenesis. The recent finding that FGFR1 mutations are involved in an autosomal dominant form of Kallmann syndrome (KAL-2), combined to the analysis of mutant mouse embryos that no longer express Fgfr1 in the telencephalon, suggests that the disease results from a deficiency in FGF signaling at the earliest stage of olfactory bulb morphogenesis. We propose that the role of the KAL1 gene product, the extracellular matrix protein anosmin-1, is to enhance FGF-signaling, and suggest that the gender difference in anosmin-1 dosage (because KAL1 partially escapes X-inactivation) explains the higher prevalence of the disease in males. PMID- 15361348 TI - [Lysophosphatidic acid: a "bioactive" phospholipid]. AB - Lysophosphatidic acid (LPA) is a "bioactive" phospholipid able to generate growth factor-like activities in a wide variety of normal and malignant cell types. LPA is proposed to play an important role in normal physiological situations such as wound healing, vascular tone, vascular integrity, or reproduction. In parallel, LPA could also be involved in the etiology of some diseases such as atherosclerosis, cancer, or obesity. The bioactivity of LPA is mediated by the activation of specific G-protein coupled receptors (LPA1, LPA2, and LPA3) leading to the activation of a number of intracellular effectors. LPA is present in solution (bound to albumin) in various extracellular fluids (blood, ascites, aqueous humor), and is released in vitro by some cell types such as platelets, cancer cells, or adipocytes. LPA is a rather polar phospholipid, which cannot easily diffuse throughout plasma membrane, and its presence outside the cells requires soluble phospholipases (secreted phospholipase A2 and soluble lysophospholipase D/autotaxin), which synthesize LPA directly in the extracellular milieu, from precursors such as phosphatidic acid and lysophosphatidylcholine. In the future, LPA receptors, as well as the enzymes involved in LPA metabolism, will constitute promising pharmacological and transgenic targets to determine the physiopathological relevance of "bioactive" LPA in vivo. PMID- 15361349 TI - [Bibliography search in biology and medicine: a tutorial for Medline PubMed use]. AB - To be informed about the last publications recently published or to produce a bibliography in a given thematic field is essential for researchers in the biomedical field. If the use of Internet information searching tools such as "Google" or "Alltheweb" makes possible to discover a great part of the grey literature, bibliographic databases like Embase, Current Contents, Biosis or Medline via PubMed are essential tools to locate scientific articles. Among these bibliographic databases, Medline PubMed, thanks to its free access, is the most used. However, a correct utilization of the various functionalities proposed (thesaurus MeSH, systematization of bibliographic searches...), and consequently the quality of bibliographic researches carried out in this database, requires to master elementary knowledge which are exposed in this article. PMID- 15361350 TI - [Economic approaches to smoking]. AB - In order to assess regulator's legitimacy in discouraging people from smoking, economists model tobacco addiction. In this paper, we review the extent economic literature about tobacco consumption and tobacco addiction. Two main approaches are singled out. In the first section, we detail the main characteristics of the two main theoretical models : rational addiction on one hand, which stylises individual choosing now to give up their future liberty to choose whether or not to smoke; and health capital on the other hand, following which individuals trade off between current pleasure and life expectancy. Second section gives account of the empirical tests of these two theories, before a last section briefly discusses and concludes. We show that the health capital theory seems to pass the test better than the rational addiction model. It is important to discriminate between these two models, from a political economy point of view. Were rational addiction true, a constant and strong increase of taxes on tobacco would be the soundest policy to curb consumption. On the contrary, if smokers are trapped into addiction and can't stop without pain, then tax increase is imposing too much strain on them, for no result. An efficient policy would be to subsidise quitting programmes, or to help smokers reducing their consumption through partial prohibition (in public transports for instance). PMID- 15361351 TI - [RNA interference: towards a functional genomics in mammalian cells?]. AB - The discovery of the induction of RNA degradation by double stranded RNA in C. elegans, "RNA interference", makes it possible to envision systematic studies of gene function in mammalian cells. Indeed, in spite of the existence in mammals of the interferon response to double stranded RNA, the introduction of small interfering RNA can induce a sequence specific inhibition of gene expression either through RNA degradation or by blocking translation. Although the inhibition is transient and usually not complete, strategies have been developed to achieve long term gene silencing. The issue of target specificity is still not completely clear and will probably constitute a limitation of this approach. However, because of the unprecedented ease with which large scale screens can be performed, RNA interference has already established itself as the tool of choice to initiate functional genomics in mammalian cells. PMID- 15361352 TI - [A technical frontier is not the answer to an ethical problem]. PMID- 15361353 TI - [Proposals and recommendations of the "Conseil scientifique de l'Inserm" in the perspective of a Research guidance and programming law]. PMID- 15361354 TI - Cholesterol diet enhances daily rhythm of Pai-1 mRNA in the mouse liver. AB - Myocardial infarction frequently occurs in the morning, a phenomenon in part resulting from the downregulation of fibrinolytic activity. Plasminogen activator inhibitor-1 (PAI-1) is a key factor behind fibrinolytic activity, and its gene expression is controlled under the circadian clock gene in the mouse heart and liver. Hypercholesterolemia has been associated with impaired fibrinolysis due to enhanced PAI-1 activity, which has also been implicated in atherosclerosis. The aim of this study was to decipher whether the Pai-1 gene is still expressed daily with hypercholesterolemia. Hypercholesterolemia (1% cholesterol diet) did not significantly affect the daily expression of clock genes (Per2 and Bmal1) and clock-controlled genes (Dbp and E4bp4) in the liver (P > 0.05); however, daily expression of the Pai-1 gene and Pai-1 promoter regulating factor genes such as Nr4a1 was significantly upregulated (P < 0.01). Daily restricted feeding for 4 h during the day reset the gene expression of Per2, Pai-1, Nr4a1, and Tnf-alpha. Lesion of the suprachiasmatic nucleus, the location of the main clock system, led to loss of Per2 and Pai-1 daily expression profiles. In the present experiments, we demonstrated that hypercholesterolemia enhanced daily expression of the Pai-1, Tnf-alpha, and Nr4a1 genes in the mouse liver without affecting clock and clock controlled genes. Therefore, the risk or high frequency of acute atherothrombotic events in the morning still seems to be a factor that may be augmented under conditions of hypercholesterolemia. PMID- 15361355 TI - Short-term resistance to diet-induced obesity in A/J mice is not associated with regulation of hypothalamic neuropeptides. AB - To investigate the mechanisms underlying long-term resistance of the A/J mouse strain to diet-induced obesity, we studied, over a period of 4 wk, the expression of uncoupling proteins in brown adipose tissue and the expression of hypothalamic neuropeptides known to regulate energy homeostasis and then used microarray analysis to identify other potentially important hypothalamic peptides. Despite increased caloric intake after 2 days of high-fat feeding, body weights of A/J mice remained stable. On and after 1 wk of high-fat feeding, A/J mice adjusted their food intake to consume the same amount of calories as mice fed a low-fat diet; thus their body weight and insulin, corticosterone, free fatty acid, and glucose levels remained unchanged for 4 wk. We found no changes in hypothalamic expression of several orexigenic and/or anorexigenic neuropeptides known to play an important role in energy homeostasis for the duration of the study. Uncoupling protein-2 mRNA expression in brown adipose tissue, however, was significantly upregulated after 2 days of high-fat feeding and tended to remain elevated for the duration of the 4-wk study. Gene array analysis revealed that several genes are up- or downregulated in response to 2 days and 1 wk of high-fat feeding. Real time PCR analysis confirmed that expression of the hypothalamic IL-1 pathway (IL 1beta, IL-1 type 1 and 2 receptors, and PPM1b/PP2C-beta, a molecule that has been implicated in the inhibition of transforming growth factor-beta-activated kinase 1-mediated IL-1 action) is altered after 2 days, but not 1 wk, of high-fat feeding. The role of additional molecules discovered by microarray analysis needs to be further explored in the future. PMID- 15361356 TI - Metabolic pathways implicated in the kinetic impairment of muscle glutamine homeostasis in adult and old glucocorticoid-treated rats. AB - An impairment of muscle glutamine metabolism in response to dexamethasone (DEX) occurs with aging. To better characterize this alteration, we have investigated muscle glutamine release with regard to muscle glutamine production (net protein breakdown, de novo glutamine synthesis) in adult and old glucocorticoid-treated rats. Male Sprague-Dawley rats (3 or 24 mo old) were divided into seven groups: three groups received 1.5 mg/kg of DEX once a day by intraperitoneal injection for 3, 5, or 7 days; three groups were pair fed to the three treated groups, respectively; and one control group of healthy rats was fed ad libitum. Muscle glutamine synthetase activity increased earlier in old rats (day 3) than in adult rats (day 7), whereas an increase in muscle glutamine release occurred later in old rats (day 5) than in adult DEX-treated rats (day 3). Consequently, muscle glutamine concentration decreased later in old rats (day 5) than in adults (day 3). Finally, net muscle protein breakdown increased only in old DEX-treated rats (day 7). In conclusion, the impairment of muscle glutamine metabolism is due to a combination of an increase in glutamine production and a delayed increase in glutamine release. PMID- 15361357 TI - TGF-beta 1 downregulates CFTR expression and function in nasal polyps of non-CF patients. AB - Nasal polyposis is a chronic inflammatory disease of the upper airways. It has been suggested that ion transports and CFTR expression could be modified in epithelial cells from nasal polyps of non-cystic fibrosis patients. We compared human nasal epithelial cells from nasal polyps (NP) with control nasal mucosa (CM). The level of CFTR mRNA was studied by Northern blot analysis and protein expression was studied by immunoprecipitation both ex vivo and in vitro in primary cultures of human nasal epithelial cells at the air-liquid interface. Ion transports were evaluated by short-circuit measurements in vitro. CFTR gene and protein expressions were significantly decreased in NP native tissues and in culture on day 4, when a global defect of ion transports was observed in NP cultures, but not in CM. We evaluated the effect of transforming growth factor (TGF)-beta 1 on CFTR expression and function in NP cultures on day 14 and showed, for the first time, that TGF-beta 1 was able to significantly downregulate the level of CFTR mRNA and cAMP-dependent current in NP cultures. Finally, we showed that the effects of TGF-beta 1 on ion transports could be reversed after 48-h removal of TGF-beta1 in NP cultures. In conclusion, our data strongly suggest that chronic inflammation in nasal polyposis downregulates CFTR gene and protein expression. PMID- 15361358 TI - CXCR2 is essential for maximal neutrophil recruitment and methacholine responsiveness after ozone exposure. AB - Ozone (O(3)), a common air pollutant, induces airway inflammation and airway hyperresponsiveness. In mice, the neutrophil chemokines KC and macrophage inflammatory protein-2 (MIP-2) are expressed in the lungs following O(3) exposure. The purpose of this study was to determine whether CXCR2, the receptor for these chemokines, is essential to O(3)-induced neutrophil recruitment, injury to lungs, and increases in respiratory system responsiveness to methacholine (MCh). O(3) exposure (1 ppm for 3 h) increased the number of neutrophils in the bronchoalveolar lavage fluid (BALF) of wild-type (BALB/c) and CXCR2-deficient mice. However, CXCR2-deficient mice had significantly fewer emigrated neutrophils than did wild-type mice. The numbers of neutrophils in the blood and concentrations of BALF KC and MIP-2 did not differ between genotypes. Together, these data suggest CXCR2 is essential for maximal chemokine-directed migration of neutrophils to the air spaces. In wild-type mice, O(3) exposure increased BALF epithelial cell numbers and total protein levels, two indirect measures of lung injury. In contrast, in CXCR2-deficient mice, the number of BALF epithelial cells was not increased by O(3) exposure. Responses to inhaled MCh were measured by whole body plethysmography using enhanced pause as the outcome indicator. O(3) exposure increased responses to inhaled MCh in both wild-type and CXCR2-deficient mice 3 h after O(3) exposure. However, at 24 h after exposure, responses to inhaled MCh were elevated in wild-type but not CXCR2-deficient mice. These results indicate CXCR2 is essential for maximal neutrophil recruitment, epithelial cell sloughing, and persistent increases in MCh responsiveness after an acute O(3) exposure. PMID- 15361359 TI - Effects of dexamethasone on Muc5ac mucin production by primary airway goblet cells. AB - Mucus hypersecretion associated with airway inflammation is reduced by glucocorticoids. Two mechanisms of glucocorticoid-mediated inhibition of mucus production have been proposed, direct inhibition of mucus production by airway epithelial cells and indirectly through inhibition of proinflammatory mediators that stimulate mucus production. In this study, we examined the effect of dexamethasone (DEX) on mRNA expression and synthesis of MUC5AC by A549 human lung adenocarcinoma cells as well as Muc5ac and total high-molecular-weight (HMW) mucins by primary rat tracheal surface epithelial (RTSE) cells. Our results showed that in primary RTSE cells, DEX 1) dose dependently suppressed Muc5ac mRNA levels, but the levels of cellular Muc5ac protein and HMW mucins were unaffected; 2) did not affect constitutive or UTP-stimulated mucin secretion; 3) enhanced the translation of Muc5ac; and 4) increased the stability of intracellular Muc5ac protein by a mechanism other than the inhibition of the proteasomal degradation. In A549 cells, however, DEX suppressed both MUC5AC mRNA levels and MUC5AC protein secretion in a dose-dependent manner. We conclude that whereas DEX inhibits the levels of Muc5ac mRNA in primary RTSE cells, the levels of Muc5ac protein remain unchanged as a consequence of increases in both translation and protein stability. Interestingly, some of the effects of DEX were opposite in a cell line. PMID- 15361360 TI - Hypoxia-induced dysfunction of rat diaphragm: role of peroxynitrite. AB - Oxidants may play a role in hypoxia-induced respiratory muscle dysfunction. In the present study we hypothesized that hypoxia-induced impairment in diaphragm contractility is associated with elevated peroxynitrite generation. In addition, we hypothesized that strenuous contractility of the diaphragm increases peroxynitrite formation. In vitro force-frequency relationship, isotonic fatigability, and nitrotyrosine levels were assessed under hypoxic (Po(2) approximately 6.5 kPa) and hyperoxic (Po(2) approximately 88.2 kPa) control conditions and also in the presence of authentic peroxynitrite (60 min), ebselen (60 min), and the nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine acetate (L-NMMA) (90 min). A hypoxia-induced downward shift of the force frequency relationship was associated with elevated nitrotyrosine level in the diaphragm. During hypoxia, both ebselen and L-NMMA decreased nitrotyrosine levels but did not affect force generation. Strenuous contractions impaired force generation but did not affect nitrotyrosine levels in the diaphragm during hypoxia. But under hyperoxic conditions, fatiguing contractions were associated with elevated diaphragm nitrotyrosine levels. Under hyperoxic conditions exogenous peroxynitrite impaired force generation and increased nitrotyrosine level. These studies show that hypoxia-induced impairment in diaphragm contractility is associated with increased diaphragm protein nitration, but no causal relationship was found between diaphragm nitrotyrosine formation and in vitro force generation. PMID- 15361361 TI - Multihormonal regulation of hepatic sinusoidal Ntcp gene expression. AB - Bile acids are efficiently removed from sinusoidal blood by a number of transporters including the Na+-taurocholate-cotransporting polypeptide (Ntcp). Na+-dependent bile salt uptake, as well as Ntcp, are expressed twofold higher in male compared with female rat livers. Also, estrogen administration to male rats decreases Ntcp expression. The aims of this study were to determine the hormonal mechanism(s) responsible for this sexually dimorphic expression of Ntcp. We examined castrated and hypophysectomized rats of both sexes. Sex steroid hormones, growth hormone, thyroid, and glucocorticoids were administered, and livers were examined for changes in Ntcp messenger RNA (mRNA). Ntcp mRNA and protein content were selectively increased in males. Estradiol selectively decreased Ntcp expression in males, whereas ovariectomy increased Ntcp in females, confirming the importance of estrogens in regulating Ntcp. Hypophysectomy decreased Ntcp mRNA levels in males and prevented estrogen administration from decreasing Ntcp, indicating the importance of pituitary hormones. Although constant infusion of growth hormone to intact males reduced Ntcp, its replacement alone after hypophysectomy did not restore the sex differences. In contrast, thyroid hormone and corticosterone increased Ntcp mRNA in hypophysectomized rats. Sex differences in Ntcp mRNA levels were produced only when the female pattern of growth hormone was administered to animals also receiving thyroid and corticosterone. Thyroid and dexamethasone also increased Ntcp mRNA in isolated rat hepatocytes, whereas growth hormone decreased Ntcp. These findings demonstrate the essential role that pituitary hormones play in the sexually dimorphic control of Ntcp expression in adult rat liver and in the mediation of estrogen effects. PMID- 15361362 TI - Effect of lower esophageal sphincter tone and crural diaphragm contraction on distensibility of the gastroesophageal junction in humans. AB - Previous studies of distensibility of the gastroesophageal junction (GEJ) in humans have not tried to distinguish between the effects of muscle action and passive elastic tissue properties of the GEJ. We studied 15 healthy subjects (ages 23-67 yr, 11 men/4 women) by using a catheter with a highly complaint bag positioned manometrically at the GEJ. The bag was distended with air at a rate of 20 ml/min while intrabag pressure was recorded. Distensions were performed during normal breathing, with breath held at maximum inspiration (MI) to activate the diaphragmatic crura, and with midesophageal balloon distension (BD) to relax the lower esophageal sphincter. In 10 subjects, distensions were performed after atropine injection (12 microg/kg iv). Pressure-volume curves and incremental distensibility values were calculated and compared among the different conditions. Both MI and BD significantly altered the slopes of the pressure volume curves, whereas no effect was seen with atropine. Maximum distensibility was seen at the volume increment of 5-10 ml and was reduced with larger volumes. Distensibility measurements for the various test conditions tended to converge at the largest volume increment, suggesting that distensibility at this degree of distension was more related to the passive elastic properties of the GEJ. On the basis of these findings, we conclude that there can be significant active muscular contributions to recordings of distensibility at the GEJ, variations that must be controlled for during different study conditions. PMID- 15361363 TI - Mechanics and hemodynamics of esophageal varices during peristaltic contraction. AB - Our hypothesis states that variceal pressure and wall tension increase dramatically during esophageal peristaltic contractions. This increase in pressure and wall tension is a natural consequence of the anatomy and physiology of the esophagus and of the esophageal venous plexus. The purpose of this study was to evaluate variceal hemodynamics during peristaltic contraction. A simultaneous ultrasound probe and manometry catheter was placed in the distal esophagus in nine patients with esophageal varices. Simultaneous esophageal luminal pressure and ultrasound images of varices were recorded during peristaltic contraction. Maximum variceal cross-sectional area and esophageal luminal pressures at which the varix flattened, closed, and opened were measured. The esophageal lumen pressure equals the intravariceal pressure at variceal flattening due to force balance laws. The mean flattening pressures (40.11 +/- 16.77 mmHg) were significantly higher than the mean opening pressures (11.56 +/- 25.56 mmHg) (P < or = 0.0001). Flattening pressures >80 mmHg were generated during peristaltic contractions in 15.5% of the swallows. Variceal cross sectional area increased a mean of 41% above baseline (range 7-89%, P < 0.0001) during swallowing. The peak closing pressures in patients that experience future variceal bleeding were significantly higher than the peak closing pressures in patients that did not experience variceal bleeding (P < 0.04). Patients with a mean peak closing pressure >61 mmHg were more likely to bleed. In this study, accuracy of predicting future variceal bleeding, based on these criteria, was 100%. Variceal models were developed, and it was demonstrated that during peristaltic contraction there was a significant increase in intravariceal pressure over baseline intravariceal pressure and that the peak intravariceal pressures were directly proportional to the resistance at the gastroesophageal junction. In conclusion, esophageal peristalsis in combination with high resistance to blood flow through the gastroesophageal junction leads to distension of the esophageal varices and an increase in intravariceal pressure and wall tension. PMID- 15361364 TI - Pleiomorphic adenoma gene-like-2, a zinc finger protein, transactivates the surfactant protein-C promoter. AB - Expression of surfactant protein (SP)-C occurs principally in type II pneumocytes located in the distal lung alveolae. SP-C expression is thought to be primarily regulated by thyroid transcription factor (TTF)-1 and its associated proteins interacting with a previously defined promoter region between -197 and -158 in mice. We screened a human lung cDNA library using a modified yeast one-hybrid system and identified pleiomorphic adenoma gene-like (PLAGL)-2, a ubiquitously expressed zinc finger protein, as a transfactor of the SP-C promoter. The PLAGL2 DNA-binding site was located in the SP-C promoter proximal region close to the TTF-1 sites. This site was demonstrated to be functional by use of electrophoresis mobility shift assay, mutagenesis analysis, and transfection studies. PLAGL2 bound to DNA via its N-terminus zinc fingers and activated the SP C promoter in a TTF-1-independent manner. Both human and mouse SP-C promoters, but not the SP-B promoter, could be activated by PLAGL2 in transfected human embryonic kidney-293 (HEK293) cells as well as in murine type II (MLE12) cells. The expression of PLAGL2 in isolated human embryonic lung type II cells and its transactivation activity on the SP-C promoter suggest that PLAGL2 may modulate SP C expression during lung development. PMID- 15361365 TI - Increased levels and activity of matrix metalloproteinase-9 in obstructive sleep apnea syndrome. AB - Matrix metalloproteinases (MMPs) are involved in the pathogenesis of cardiovascular diseases. We examined serum levels of MMP-9 and its inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), activity of MMP-9, and the effect of nasal continuous positive airway pressure (nCPAP) in patients with obstructive sleep apnea syndrome (OSAS). After polysomnography, venous blood was collected at 5:00 A.M. from 44 patients with OSAS and 18 control subjects who were obese, and serum levels of MMP-9, TIMP-1, and enzymatic activity of MMP-9 were measured. In addition, the effects of 1 month of treatment with nCPAP were studied in patients with moderate to severe OSAS. Although serum levels of MMP-9 (p < 0.03) and MMP-9 activity (p < 0.01) were higher in patients with OSAS than in control subjects who were obese, TIMP-1 levels did not differ significantly. In patients with OSAS, the severity of OSAS was the primary factor influencing levels (p < 0.01) and activity (p < 0.01) of MMP-9. nCPAP significantly decreased serum levels (p < 0.01) and activity (p < 0.001) of MMP-9 but did not affect TIMP 1 levels. Therefore, OSAS may increase risks of cardiovascular morbidity, and nCPAP might be useful for decreasing these risks. PMID- 15361366 TI - Azithromycin blocks neutrophil recruitment in Pseudomonas endobronchial infection. AB - Macrolides exert their effects on the host by modulation of immune responses. In this study, we assessed the therapeutic efficacy of azithromycin in a murine model of mucoid Pseudomonas aeruginosa endobronchial infection. The clearance of Pseudomonas from the airway of mice treated with the macrolide azithromycin was not different than untreated mice challenged with Pseudomonas beads. However, the azithromycin-treated mice showed a remarkable reduction in lung cellular infiltrate in response to Pseudomonas beads, as compared with untreated mice. This effect was associated with significant decreases in lung levels of tumor necrosis factor-alpha and keratinocyte-derived chemokine in azithromycin-treated mice compared with untreated mice. Furthermore, there was a significant reduction in the response of both mouse and human neutrophils to chemokine-dependent and independent chemoattractants when studied in vitro. Inhibition of chemotaxis correlated with azithromycin-mediated inhibition of extracellular signal regulated kinase-1 and -2 activation. This study indicates that the azithromycin treatment in vivo results in significant reduction in airway-specific inflammation, which occurs in part by inhibition of neutrophil recruitment to the lung through reduction in proinflammatory cytokine expression and inhibition of neutrophil migration via the extracellular signal-regulated kinase-1 and -2 signal transduction pathway. PMID- 15361367 TI - Lower respiratory viral illnesses: improved diagnosis by molecular methods and clinical impact. AB - We assessed the frequency and the potential role of respiratory viruses on disease outcomes in hospitalized patients and lung transplant recipients who underwent a bronchoalveolar lavage (BAL) for an acute respiratory infection. BAL specimens (148) were analyzed by reverse transcription-polymerase chain reaction for the presence of 11 different viruses, as well as Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila. Respiratory viruses were identified in 34 of 117 BAL specimens (29%) obtained in patients with a suspected respiratory infection and in only 2 of 31 control subjects (7%) (p < 0.01). M. pneumoniae was identified in five additional cases. Only 30% of cases that were virus positive by molecular methods were also positive by cell culture analysis. Rhinovirus was the most frequently identified virus (56% of cases) followed by respiratory syncytial virus (27%). In lung transplant recipients, the rate of viral infections was 55% in cases with respiratory symptoms compared with only 4% in control subjects (p < 0.001). In these cases, respiratory viral infections were associated with significant lung function abnormalities. By using reverse transcription-polymerase chain reaction assays, we frequently identified respiratory viruses in BAL specimens of patients hospitalized with lower respiratory tract infections. These viruses were associated with high morbidity, particularly in lung transplant recipients. PMID- 15361368 TI - Impaired transforming growth factor-beta signaling in idiopathic pulmonary arterial hypertension. AB - Mutations in transforming growth factor-beta family receptor-II, bone morphogenetic protein receptor-2, and activin-like kinase-1 have been associated with pulmonary hypertension. In the present study, we determined that pulmonary arteries in normal lungs and in lungs of patients with emphysema and idiopathic pulmonary arterial hypertension comparably expressed transforming growth factor beta receptors I and II, Smad(1, 5, 8), Smad2, Smad3, Smad4, phosphorylated Smad(1, 5, 8), and phosphorylated Smad2 (the latter two both indicative of active in vivo signaling) in endothelial cells, as assessed by immunohistochemistry and quantitative morphometry. Medial or intimal smooth muscle cells had weak or absent expression of these molecules. In clear contrast to endothelial cell expression in pulmonary arteries and in endothelial cells lining incipient vessels within plexiform lesions of hypertensive lungs, endothelial cells present in the core of the lesions lacked expression of all examined members of the signaling molecules. These findings were made irrespective of the mutation status of bone morphogenetic protein receptor-2 in hypertensive patients. Our findings suggest that pulmonary artery endothelial cells in both normal and severely hypertensive lungs have active transforming growth factor-beta family signaling, and that loss of signaling might contribute to the abnormal growth of endothelial cells in plexiform lesions in idiopathic pulmonary arterial hypertension. PMID- 15361369 TI - Urinary bladder cancer in Wegener's granulomatosis: is it more than cyclophosphamide? PMID- 15361370 TI - Case number 31: Nodular panniculitis as expression of Munchausen's syndrome (panniculitis artefacta). PMID- 15361371 TI - EP2/EP4 signalling inhibits monocyte chemoattractant protein-1 production induced by interleukin 1beta in synovial fibroblasts. AB - BACKGROUND: Besides its proinflammatory properties, prostaglandin E(2) (PGE(2)) acts as a regulator of the expression of inducible genes. Inhibition of PGE(2) synthesis might thus result in a paradoxical deleterious effect on inflammation. OBJECTIVE: To examine the effect of PGE(2) on monocyte chemoattractant protein-1 (MCP-1) expression in cultured synovial fibroblasts (SF) stimulated with interleukin (IL)1beta. METHODS: MCP-1 expression was assessed in SF stimulated with IL1beta in the presence of PGE(2) or different NSAIDs by RT-PCR or northern blot and immunocytochemistry. Expression of cyclo-oxygenase (COX) isoforms was studied by western blot techniques. The role of PGE(2) receptors (EP) in PGE(2) action was assessed employing EP receptor subtype-specific agonists. RESULTS: PGE(2) significantly inhibited IL1beta induced MCP-1 expression in SF in a dose dependent manner. IL1beta increased COX-2 and did not alter COX-1 synthesis in SF. 11-Deoxy-PGE(1), an EP(2)/EP(4) agonist, reproduced PGE(2) action on MCP-1 expression. Butaprost, a selective EP(2) agonist, was less potent than PGE(2). Sulprostone, an EP(1)/EP(3) agonist, had no effect on IL1beta induced MCP-1 expression. Inhibition of endogenous PGE(2) synthesis by NSAIDs further enhanced MCP-1 mRNA expression in IL1beta stimulated SF, an effect prevented by addition of exogenous PGE(2). CONCLUSION: Activation of EP(2)/EP(4) receptors down regulates the expression of MCP-1 in IL1beta stimulated SF, while PGE(2) pharmacological inhibition cuts off this signalling pathway and results in a superinduction of MCP-1 expression. The data suggest that NSAIDs may intercept a natural regulatory circuit controlling the magnitude of inflammation, which questions their continuous administration in inflammatory joint diseases. PMID- 15361372 TI - Discrepancy between mRNA and protein expression of tumour suppressor maspin in synovial tissue may contribute to synovial hyperplasia in rheumatoid arthritis. AB - OBJECTIVE: To investigate the expression of maspin in RA synovial tissue and compare it with the expression in osteoarthritis (OA) and normal synovial tissue (NS). METHODS: Using specific primers for maspin, a 237 bp fragment was amplified from cDNA obtained from cultured RA, OA, and normal synovial fibroblasts (SF) by RT-PCR. Additionally, mRNA expression levels were determined quantitatively by real time PCR. mRNA expression of maspin was investigated on snap frozen and paraffin embedded synovial tissue sections by in situ hybridisation. Immunohistochemistry was used to identify the cell type expressing maspin. SDS PAGE and western blotting were performed to evaluate the protein expression in cultured SF. To confirm protein synthesis in situ, immunohistochemistry with specific anti-maspin antibodies was performed in synovial tissue sections of patients with RA. RESULTS: RT-PCR showed expression of maspin in all cDNA samples from cultured SF. Maspin mRNA was found to be decreased in RA SF twofold and 70 fold compared with OA SF and NS SF, respectively. Maspin mRNA was expressed in RA, OA, and normal synovial tissue. Importantly, maspin transcripts were also found at sites of invasion into cartilage and bone. At the protein level, maspin could be detected in RA and, less prominently, OA SF. In RA synovial tissue, maspin protein was detected in only a few synovial lining cells. CONCLUSION: Maspin is expressed intensively in RA SF at the mRNA level, but only slightly at the protein level, possibly owing to down regulation of maspin; this may contribute to the hyperplasia of synovial tissue in RA. PMID- 15361373 TI - A prospective study of pregnant patients with rheumatoid arthritis and ankylosing spondylitis using validated clinical instruments. AB - OBJECTIVE: : To analyse the disease course of patients with rheumatoid arthritis (RA) and ankylosing spondylitis (AS) during and after pregnancy by validated clinical instruments for measurement of disease activity, and assess their usefulness in pregnant patients. METHODS: Included were 10 patients with RA and 9 with AS (10 pregnancies). Clinical examination and blood/urine sampling was performed before conception, at each trimester, and weeks 6, 12, and 24 post partum. Assessment of RA was by the RA Disease Activity Index (RADAI), the 44 joint count, and the Health Assessment Questionnaire; assessment of AS by the Bath Ankylosing Spondylitis Activity Index (BASDAI), the Dougados Functional and Articular Index, and a night pain index. Common for all patients were the patient's and physician's global assessment. RESULTS: : Most patients with RA showed sustained or increased improvement of disease activity during pregnancy. Higher disease activity scores were found in the patients with AS with a frequent increase of disease activity in the second trimester and mitigation of symptoms in the third trimester. Analysis specifically for the patient's assessment of pain showed continuously higher pain scores in the patients with AS than in those with RA. Rank correlation showed good to moderate correlation between most clinical measurements and RADAI or BASDAI, respectively. Functional indices were confounded by physiological changes of late pregnancy. CONCLUSION: RA can be monitored during and after pregnancy by the swollen joint count and RADAI without interference from pregnancy related symptoms, whereas usual measures of disease activity are not always applicable in pregnant patients with AS. PMID- 15361374 TI - Decrease of anti-cyclic citrullinated peptide antibodies and rheumatoid factor following anti-TNFalpha therapy (infliximab) in rheumatoid arthritis is associated with clinical improvement. AB - OBJECTIVE: To investigate the effect of infliximab treatment on anti-cyclic citrullinated peptide antibodies (anti-CCP) and rheumatoid factor (RF) in patients with rheumatoid arthritis. METHODS: 43 patients with rheumatoid arthritis not responding to disease modifying anti-rheumatic drugs (DMARD) received intravenous infliximab at a dose of 3 mg/kg at baseline and after two and six weeks, and subsequently bimonthly, in combination with methotrexate. Serum samples were collected at baseline and at week 24. A commercial enzyme linked immunosorbent assay was used to test for anti-CCP antibodies; RF were detected using a quantitative nephelometric assay. RESULTS: At baseline, 38 of the 43 patients (88%) were positive for anti-CCP antibodies, and 41 (95%) were positive for RF. The serum titre of anti-CCP and RF decreased significantly after six months of treatment (p = 0.0001 and p<0.0001, respectively). When the patients were grouped on the basis of their clinical response to infliximab, a significant decrease in serum anti-CCP antibodies and RF was observed only in patients who had clinical improvement (ACR 20 and ACR 50). CONCLUSIONS: Anti TNFalpha treatment in rheumatoid arthritis results in a decrease in the serum titres of RF and anti-CCP antibodies in patients showing clinical improvement, suggesting that these measurements may be a useful adjunct in assessing treatment efficacy. PMID- 15361375 TI - Influence of guideline adherence on outcome in a randomised controlled trial on the efficacy of methotrexate with folate supplementation in rheumatoid arthritis. AB - OBJECTIVE: To study the influence of rheumatologists' adherence to a methotrexate guideline on efficacy and toxicity in the treatment of rheumatoid arthritis. METHODS: In a 48 week randomised controlled trial of methotrexate, comparing folates with placebo, rheumatologists were advised on methotrexate dosage using a guideline reflecting daily practice. The influence of guideline non-adherence on outcome was analysed using generalised estimating equations and survival analysis. RESULTS: In 51% of the 411 study patients the guidelines were always followed. Non-adherence resulted in lower doses of methotrexate in 25% of cases, and higher doses in 24%. The reduction in the disease activity score was significantly greater (mean -0.4; p = 0.0085) in the adherent group than in the "low dose" group; the "high dose" group did not differ from the adherent group. Dropout caused by severe adverse events did not differ between the three groups. CONCLUSIONS: There is an indication that adherence to guidelines on methotrexate dosage may benefit patients with rheumatoid arthritis by improving disease activity without increasing toxicity. For definite proof, a randomised controlled trial comparing guideline supported dosing with usual care is needed. PMID- 15361376 TI - Methotrexate related adverse effects in patients with rheumatoid arthritis are associated with the A1298C polymorphism of the MTHFR gene. AB - BACKGROUND: There is an association between C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene and methotrexate related toxicity. OBJECTIVE: To examine the relations between the recently described A1298C polymorphism of the MTHFR gene, plasma homocysteine, methotrexate toxicity, and disease activity in patients with rheumatoid arthritis. DESIGN: A cross sectional study on 93 methotrexate treated patients with rheumatoid arthritis, comprising a clinical interview and physical examination to determine disease activity and methotrexate related adverse reactions. Genotype analysis of the MTHFR gene was carried out and fasting plasma homocysteine and serum folate concentrations were measured. The data were analysed using univariate analysis. Allele and genotype distributions were compared with those of a healthy control group. RESULTS: The frequency of the 1298CC genotype (24.7%) in the rheumatoid study group was greater than expected in the general population (12.8%, p<0.001). This genotype was associated with a significantly low rate of methotrexate related side effects. The odds ratio for side effects in patients with wild type 1298AA genotype v 1298CC genotype was 5.24 (95% confidence interval, 1.38 to 20). No correlation of disease activity variables or plasma homocysteine with MTHFR A1298C and C677T polymorphisms was observed. CONCLUSIONS: 1298CC polymorphism was more common in methotrexate treated rheumatoid patients than expected in the population, and was associated with a reduction in methotrexate related adverse effects. The A1298C polymorphism of the MTHFR gene may indicate a need to adjust the dose of methotrexate given to patients with rheumatoid arthritis. PMID- 15361377 TI - Better efficacy of methotrexate given by intramuscular injection than orally in patients with rheumatoid arthritis. AB - OBJECTIVE: To compare the clinical efficacy of methotrexate and tolerance to the drug in patients with rheumatoid arthritis who were switched from intramuscular to oral administration because of a shortage of the intramuscular preparation. METHODS: 143 patients were switched from intramuscular to oral methotrexate. Of these, 47 were switched back to the intramuscular form. A multiple choice questionnaire was sent by mail to evaluate clinical and biological criteria of efficacy and tolerance. RESULTS: When methotrexate was first switched from intramuscular to oral administration, increased disease activity, exacerbation of morning pain and hand stiffness, duration of morning stiffness, increased joint pain, and increased joint swelling were observed. There was a greater frequency of gastrointestinal symptoms, but without a significant increase in liver abnormalities. When intramuscular methotrexate became available again, 47 of the 143 patients were switched back and were followed for at least three months. On average, disease manifestations were improved and side effects reduced by the switch. CONCLUSIONS: Methotrexate given intramuscularly had improved clinical efficacy with fewer side effects than given orally. Intramuscular methotrexate administration should be considered when rheumatoid arthritis remains active in spite of high dose oral methotrexate. PMID- 15361378 TI - Characteristics of patients with rheumatoid arthritis in France: a study of 1109 patients managed by hospital based rheumatologists. AB - OBJECTIVE: To describe the characteristics of rheumatoid arthritis in patients managed by hospital based rheumatologists in France. METHODS: All public and non profit private hospitals in France were invited to participate in a cross sectional study. Clinical data on the day of inclusion and health resources used for rheumatoid arthritis over the previous 12 months (treatments, medical devices, physician visits, examinations, hospital admissions, and other health professional care) were recorded. RESULTS: 1109 patients from 75 centres located throughout the country were included (846 female; mean disease duration, 10.6 years; mean age, 56.7 years). Active disease (swollen joint count > or =6, tender joint count > or =6, and two of: morning stiffness > or =45 min, C reactive protein > or =20 mg/l, erythrocyte sedimentation rate >28 mm/h) was observed in 146 patients (13.2%). Mean (SD) DAS(28) was 4.51 (1.55). Severe extra-articular manifestations were reported in 8.4%. ACR functional status was: class I, 19%; class II, 28%; class III, 31%; class IV, 22%. Comorbidity was observed in 44.9% of cases, particularly chronic pulmonary disease and coronary or peripheral vascular disease. Average AIMS2-SF dimension scores were between 4.56 and 6.18, and mean HAQ was 1.32 (0.77). Disease modifying antirheumatic drugs (DMARDs) were prescribed for 82.1% of the patients. During the previous four weeks, one DMARD was used in 62.5%, and two or more in 19.5%. Corticosteroids were prescribed in 72%. CONCLUSIONS: In a rheumatoid arthritis population managed by hospital based rheumatologists, the disease was active in 13% and severe in more than one third of cases. PMID- 15361379 TI - Antiphospholipid antibodies, systemic lupus erythematosus, and non-traumatic metatarsal fractures. AB - BACKGROUND: Stress fractures are common in military recruits and athletes and are thought to be secondary to stress and overuse. Less often they are associated with metabolic disorders such as osteoporosis, hypophosphataemia, and diabetes mellitus. OBJECTIVE: Analysis of 19 patients with systemic lupus erythematosus and antiphospholipid antibodies presenting consecutively with foot pain. All had metatarsal fractures (six bilateral) without any obvious history of trauma. RESULTS: 13 of the 19 patients had antiphospholipid syndrome. Among the whole group, 13 had normal bone mineral density, one had osteopenia, and five others had osteoporosis as defined by WHO criteria; 10 had received steroids, mostly in low dosage; 13 were receiving warfarin. There was no evidence that a metabolic bone abnormality was a unifying factor in the pathogenesis. CONCLUSIONS: Atraumatic metatarsal stress fractures may occur in SLE, particularly in association with the antiphospholipid syndrome. The pathogenesis of these fractures remains uncertain but microinfarcts in the metatarsal bones are a possible cause. PMID- 15361380 TI - Association of the oestrogen receptor alpha gene polymorphisms with disease onset in systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the genetic influence of PvuII and XbaI polymorphisms of oestrogen receptor alpha (ORalpha) in patients with systemic lupus erythematosus (SLE) in Korea. METHODS: Genomic DNA from 268 female controls and 137 female SLE patients (41 childhood onset and 96 adult onset) were analysed using PvuII and XbaI restriction fragment length polymorphism. Comparison of the frequencies of alleles and genotypes was made in control and patient groups and in childhood onset and adult onset SLE subgroups. RESULTS: Although the Pp genotype occurred more often in SLE patients than in controls (p(c) = 0.017), ORalpha genotype distributions of adult onset SLE did not differ significantly from controls. The PP, Pp, and xx genotypes occurred less often in childhood onset SLE (p(c) = 0.0045, 0.0498, and 0.0255, respectively) than in controls. Additionally, the PP genotype was less common in childhood onset than in adult onset SLE (p(c) = 0.016). SLE patients with the PP genotypes were older at disease onset than those with the other genotypes (p = 0.001). Patients with the Xx genotype had an earlier onset of SLE than those with the xx genotype (p = 0.025). The frequency of the combined ppXx genotype was greater in childhood onset SLE than in controls (p(c) = 0.0009) or adult onset SLE (p(c) = 0.027). The same trend was supported by subgroup analyses according to age at menarche and logistic multivariate analyses. CONCLUSIONS: ORalpha polymorphisms are significantly associated with the age at disease onset in Koreans with SLE. PMID- 15361381 TI - Anti-telomere antibodies in systemic lupus erythematosus (SLE): a comparison with five antinuclear antibody assays in 430 patients with SLE and other rheumatic diseases. AB - OBJECTIVE: To investigate the prevalence and diagnostic significance of antibodies against telomeric DNA in systemic lupus erythematosus (SLE) and other autoimmune rheumatic diseases, and to make comparisons with five conventional anti-DNA or anti-nuclear antibody (ANA) assays. METHODS: Antibodies to telomeres, which are highly repetitive sequences of DNA (TTAGGG/CCCTAA) at the end of eukaryotic chromosomes, were measured by an enzyme linked immunosorbent assay (ELISA) in 305 patients with SLE and 125 patients with other autoimmune rheumatic diseases (78 rheumatoid arthritis, 32 primary Sjogren's syndrome, eight mixed connective tissue disease, seven miscellaneous rheumatic diseases). Other assays used were two commercial ELISA assays for anti-dsDNA using calf thymus as antigen, Crithidialuciliae immunofluorescence, and radioimmunoassay (RIA) for anti-dsDNA and immunofluorescence using Hep-2 cells for ANA. RESULTS: The prevalence of anti-telomere in SLE was 60%, v 5% in rheumatoid arthritis and 18% in other autoimmune rheumatic diseases. Specificity of anti-telomere for SLE was 91%; positive and negative predictive values were 95% and 46%, respectively. For anti-dsDNA by two ELISA assays using calf thymus as antigen, sensitivities were 69% and 29% and specificities 66% and 96%, respectively. Other anti-dsDNA assays had low sensitivities (RIA 43%, Crithidia immunofluorescence 13%). The association of anti-telomere with a history of nephritis in patients with SLE was stronger (p = 0.005) than by any other assay (p = 0.006-0.999). The correlations between the different assays were good (p<0.001 for all comparisons). CONCLUSIONS: The new ELISA for anti-telomere antibodies using standardised human dsDNA as antigen is a sensitive and highly specific test for SLE. PMID- 15361383 TI - A randomised controlled trial of intra-articular corticosteroid injection of the carpometacarpal joint of the thumb in osteoarthritis. AB - OBJECTIVE: To investigate the efficacy of corticosteroid injections into the carpometacarpal joint of the thumb (CMCJ) in patients with osteoarthritis. DESIGN: A double blind, randomised controlled trial using 40 hospital referred patients with CMCJ osteoarthritis who received intra-articular injections of 5 mg triamcinolone hexacetonide (0.25 ml) or sterile 0.9% saline (0.25 ml). Injections were given under imaging control. MAIN OUTCOME MEASURES: The primary outcome was improvement in a pain visual analogue score (VAS) of 20% at 24 weeks. In addition patients were assessed at 4, 12, and 24 weeks for joint stiffness, joint tenderness, and physician and patient global assessments. Hand radiographs were evaluated for the degree of CMC joint space narrowing and marginal osteophytes according to the OARSI atlas. RESULTS: Baseline clinical variables were not significantly different between the two treatment groups. There was no improvement in the VAS of pain at 24 weeks. At each assessment point there was no significant difference between the steroid and placebo groups in median values for joint stiffness, joint tenderness, or patient and physician global assessments. Non-parametric analysis of each group individually revealed statistically significant improvements in patient and physician global assessments at weeks 4, 12, and 24 in the placebo group and at weeks 4 and 12 in the steroid group. CONCLUSIONS: No clinical benefit was gained from intra articular steroid injection to the CMCJ in moderate to severe osteoarthritis compared with placebo injection. PMID- 15361382 TI - Genetic mechanisms of knee osteoarthritis: a population based case-control study. AB - OBJECTIVE: To compare subjects who had at least one parent with a total knee replacement for severe primary knee osteoarthritis with age and sex matched controls who had no family history of knee osteoarthritis DESIGN: Population based case-control study of 188 matched pairs (mean age 45 years, range 26 to 60). METHODS: Articular cartilage volume and bone size were determined at the patella and at the medial tibial and lateral tibial compartments by processing images acquired using T1 weighted, fat saturated magnetic resonance imaging. Radiographic osteoarthritis (ROA) was assessed from a standing semiflexed radiograph scored for joint space narrowing and osteophytosis. Knee pain was assessed by questionnaire. Height, weight, body mass index (BMI), lower limb muscle strength, and endurance fitness were measured by standard protocols. RESULTS: Compared with the controls, index offspring had higher BMI (27.8 v 26.0 kg/m(2), p = 0.02), weaker lower limb muscles (127 v 135 kg, p = 0.006), more knee pain (47% v 22%, p<0.001), and greater medial tibial bone area (17.6 v 17.1 cm(2), p = 0.01). With the exception of BMI, these differences persisted in multivariate analysis. There was a non-significant trend to higher cartilage volume at tibial sites and increased ROA in the offspring in the total and subgroup analyses, but no difference in height and endurance fitness. CONCLUSIONS: BMI, muscle strength, knee pain, and medial tibial bone area, but not cartilage volume, appear to play a role in the genetic regulation and development of knee osteoarthritis. PMID- 15361384 TI - Is avoidant coping independent of disease status and stable over time in patients with ankylosing spondylitis? AB - OBJECTIVE: To determine whether avoidant coping in ankylosing spondylitis (AS) is independent of disease status and whether it is stable over time. METHODS: 658 patients with AS completed a postal questionnaire on health status, including pain and stiffness (BASDAI), physical function (BASFI), and coping (CORS). In CORS, "decreasing activities to cope with pain" and "pacing to cope with limitations" reflect avoidant behavioural coping. Ninety patients continued in a longitudinal study and 70 completed the CORS after four years. The adjusted contribution of age, sex, disease duration, educational level, pain (BASDAI), and physical function (BASFI) to the two avoidant coping strategies at first assessment was determined by multiple linear regression. Agreement between coping at first assessment and four years later was determined by intraclass correlation, and the correlation between change in coping and change in disease status over time by Pearson's correlation. RESULTS: At first assessment, worse physical function (BASFI) and more pain (BASDAI) were associated with "decreasing activities to cope with pain". Worse physical function, but not pain, was associated with "pacing to cope with limitations". The contribution of physical function or pain to the total explained variance in each of the coping strategies was small. Disease duration was not a determinant of avoidant coping, but greater age was associated with "pacing to cope with limitations". Change in avoidant coping strategies over time could not be explained by change in function or pain. CONCLUSIONS: In AS, avoidant coping at a particular time is largely independent of disease duration or status. Variability in avoidant coping over a limited period of four years cannot be explained by change in disease status. PMID- 15361385 TI - Attitudes to early rheumatoid arthritis: changing patterns. Results of a survey. AB - OBJECTIVE: To determine if rheumatologists have changed their views on diagnosis and treatment of early rheumatoid arthritis (RA). METHODS: Three consecutive questionnaires were sent out to international rheumatologists in 1997, 2000, and 2003. The following aspects of early RA were covered: definition; patient referral time; diagnostic means; follow up intervals; and treatment strategies. All initial participants who responded to at least one of the follow up surveys were included in the analysis. RESULTS: RA is now defined by a smaller number of affected joints (monarthritis: 9.8% respondents in 1997 v 17.4% in 2003), and shorter symptom duration (<3 months: 65.5% in 1997 v 85.8% in 2003). Early referrals (<6 weeks) increased (8.9% in 1997 v 17.4% in 2003). Serological test for diagnosis was mostly rheumatoid factor (100% in 2003), but anti-CCP was already used by 17.4% in 2003. Follow up of patients with early RA intensified (every 2 weeks: 16.1% in 1997 v 30.4% in 2003; every month: 47.8% in 2003 v 64.3% in 1997). Treatment with disease modifying antirheumatic drugs (DMARDs) mainly comprised methotrexate, sulfasalazine, and antimalarial drugs. Leflunomide was among the two favourite DMARDs of 10.9% in 2003, whereas no biological agent was so. In 2003, 46.7% respondents started treatment with DMARDs if RA was suspected (30.9% in 1997); no one waited for erosions to occur (7.3% in 1997). CONCLUSION: The data obtained in this study suggest that the concept of diagnosing and treating RA early is accepted by a large proportion of the rheumatological community. PMID- 15361386 TI - Increased prevalence of ocular glaucomatous abnormalities in systemic sclerosis. AB - BACKGROUND: Cardiovascular diseases, vasospasm, and dysimmunity have been implicated in normal tension glaucoma (NTG). OBJECTIVE: To investigate the prevalence of ocular abnormalities suggestive of glaucoma damage in systemic sclerosis (SSc). METHODS: 61 patients with SSc (mean (SD) age 56.2 (12) years, mean (SD) disease duration 9.9 (9) years; 41 with limited cutaneous disease) and 37 control subjects with osteoarthritis (mean (SD) age 55.9 (12) years) were studied. They were systematically referred to an ophthalmologist. The evaluation was based on aplanation tonometry, ophthalmoscopy with retinal photography (evaluation of cup/disc ratio (c/d)), and automated static perimetry (determination of mean defects (MD)). Statistical analyses were performed with the chi(2), Mann-Whitney, and Spearman tests. RESULTS: The mean visual acuity and intraocular pressure were similar in both groups. An excavation with a c/d >0.3 was found in 27 eyes from patients with SSc and 5 eyes from controls (p = 0.009); a c/d >0.7 was found in 4 eyes from patients with SSc and none in the controls (NS). Visual field defects (MD <-2 dB) were found in 55 eyes from patients with SSc and in 18 eyes from controls (p<0.0001). A concomitant c/d >0.3 and MD <-2 dB was found in 21 eyes from 12 patients with SSc but in none of the control eyes (p<0.0001). CONCLUSION: Ocular abnormalities suggesting glaucomatous neuropathy without ocular hypertension were dramatically more prevalent in patients with SSc. These abnormalities seem to be mild but justify long term follow up. They are consistent with the vascular pathogenic hypothesis for NTG. PMID- 15361387 TI - A disease activity score for polymyalgia rheumatica. AB - OBJECTIVE: To develop a composite score for measurement of disease activity in polymyalgia rheumatica (PMR) and assess its internal and external validity. METHODS: A PMR activity score (AS) was designed and assessed for internal and external validity in two patient cohorts: 57 international patients evaluated primarily for development of the PMR-AS at baseline, weeks 4 and 24; and for validation, 24 Austrian patients assessed at baseline, week 4, and at a mean (SD) point of week 33.6 (24.5). The PMR-AS was calculated as: CRP (mg/dl)+VAS p (0 10)+VAS ph (0-10)+(MST (min)x0.1)+EUL (3-0); Cronbach's alpha was calculated. Factor analysis by linear regression was applied, and responses calculated on the basis of the PMR response criteria and the PMR-AS applied. PMR-AS values at different times were compared by paired t tests. RESULTS: Cronbach's alpha for the composite score was 0.91 and 0.88 in the two cohorts. Factor analysis showed that each single item contributed significantly to the total score and the relative weight of each item in both cohorts was equally distributed. Mean PMR-AS at baseline was 27.54 and 28.72, respectively, at week 4, 5.99 and 8.99, and at the final visit 5.35 and 5.92 (NS). PMR-AS values at baseline and at later visits were significantly different (p<0.0001). PMR-AS values <7 indicated low disease activity, 7-17 medium disease activity, and >17 high PMR activity. In a third control cohort the PMR-AS correlated highly with patient's global assessment, patient satisfaction, and ESR (p<0.001). CONCLUSION: The PMR-AS provides an easily applicable and valid tool for monitoring disease activity, and in combination with the PMR response criteria provides a better description of response. PMID- 15361388 TI - Detection of small joint synovitis by ultrasonography: the learning curve of rheumatologists. AB - BACKGROUND: Ultrasonography allows assessment of soft tissue structures and has become a valued tool for diagnosing synovitis. OBJECTIVE: To assess the learning curve for ultrasonography in evaluating synovitis of the small joints in rheumatoid arthritis. METHODS: Metacarpophalangeal (MCP), metatarsophalangeal (MTP), and proximal interphalangeal (PIP) joints were evaluated using ultrasonography (Esaote AU 5 Epi, linear probe 10-13 MHz) by four rheumatologists, the first being experienced (senior), the others having no (fellows 1 and 2) or little (fellow 3) experience in ultrasonography. For each fellow, the learning curve was divided into blocks. In each block the fellow examined five consecutive patients with the senior; then, blinded to the senior's results, two further patients alone (seven patients examined per block). For each evaluation, the MCP, PIP, and MTP joints were individually tagged as having synovitis or not. The ultrasonography results were compared between fellow and senior for the two last patients of each block, using proportions of agreement and kappa statistics. RESULTS: 70 patients were evaluated (seven practice patients, followed by nine blocks). For fellows 1 and 2, the proportions of agreement were respectively 42% and 47% (kappa = 0 and 0) at the first evaluation, and rose progressively to 82% and 82% (kappa = 0.63 and 0.62) at the ninth evaluation. For fellow 3, initially good results were followed by decreased accuracy. CONCLUSIONS: Detecting synovitis of the MCP, PIP, and MTP joints using ultrasonography can be done accurately by rheumatologists initially not experienced in this technique. At least 70 examinations were necessary to develop competence. PMID- 15361389 TI - Interrater reproducibility of clinical tests for rotator cuff lesions. AB - BACKGROUND: Rotator cuff lesions are common in the community but reproducibility of tests for shoulder assessment has not been adequately appraised and there is no uniform approach to their use. OBJECTIVE: To study interrater reproducibility of standard tests for shoulder evaluation among a rheumatology specialist, rheumatology trainee, and research nurse. METHODS: 136 patients were reviewed over 12 months at a major teaching hospital. The three assessors examined each patient in random order and were unaware of each other's evaluation. Each shoulder was examined in a standard manner by recognised tests for specific lesions and a diagnostic algorithm was used. Between-observer agreement was determined by calculating Cohen's kappa coefficients (measuring agreement beyond that expected by chance). RESULTS: Fair to substantial agreement was obtained for the observations of tenderness, painful arc, and external rotation. Tests for supraspinatus and subscapularis also showed at least fair agreement between observers. 40/55 (73%) kappa coefficient assessments were rated at >0.2, indicating at least fair concordance between observers; 21/55 (38%) were rated at >0.4, indicating at least moderate concordance between observers. CONCLUSION: The reproducibility of certain tests, employed by observers of varying experience, in the assessment of the rotator cuff and general shoulder disease was determined. This has implications for delegation of shoulder assessment to nurse specialists, the development of a simplified evaluation schedule for general practitioners, and uniformity in epidemiological research studies. PMID- 15361390 TI - A comparison of four shoulder-specific questionnaires in primary care. AB - OBJECTIVES: To compare the validity, responsiveness to change, and user friendliness of four self completed, shoulder-specific questionnaires in primary care. METHODS: A cross sectional assessment of validity and a longitudinal assessment of responsiveness to change of four shoulder questionnaires was carried out: the Dutch Shoulder Disability Questionnaire (SDQ-NL); the United Kingdom Shoulder Disability Questionnaire (SDQ-UK); and two American instruments, the Shoulder Pain and Disability Index (SPADI) and the Shoulder Rating Questionnaire (SRQ). 180 primary care consulters with new shoulder region pain each completed two of the questionnaires, as well as EuroQoL and 10 cm visual analogue scales (VAS) for overall pain and difficulty due to the shoulder problem. Each participant was assessed by a standardised clinical schedule. Postal follow up at 6 weeks included baseline measures and self rated assessment of global change of the shoulder problem (seven point Likert scale). RESULTS: Strongest correlations were found for SDQ-UK with EuroQoL 5 score, and for SPADI and SRQ with shoulder pain and difficulty VAS. All shoulder questionnaires correlated poorly with active movement at the painful shoulder. SPADI and SRQ performed better on ROC analysis than SDQ-NL and SDQ-UK (areas under the curve of 0.87, 0.85, 0.77, and 0.77, respectively). However, SRQ scores changed significantly over time in stable subjects. CONCLUSIONS: Cross sectional comparison of the four shoulder questionnaires showed they had similar overall validity and patient acceptability. SPADI and SRQ were most responsive to change. Additionally, SPADI was the quickest to complete and scores did not change significantly in stable subjects. PMID- 15361391 TI - Predominance of Th1 cytokine in peripheral blood and pathological tissues of patients with active untreated adult onset Still's disease. AB - OBJECTIVE: To determine the type 1 T helper (Th1)/type 2 T helper (Th2) balance in the peripheral blood (PB) and pathological tissues of patients with active untreated adult onset Still's disease (AOSD). METHODS: The percentages of interferon gamma (IFNgamma)- and interleukin (IL)4-producing Th cells in the PB of 20 patients with active untreated AOSD, 20 patients with active rheumatoid arthritis (RA), and 20 healthy controls were determined by intracellular staining and flow cytometry. Serum levels of IL18 and soluble IL2 receptor were measured by enzyme linked immunosorbent assay. Levels of IFNgamma and IL4 messenger (m) RNA expression were examined by real time quantitative polymerase chain reaction in biopsy specimens of evanescent rash and synovitis from 8 patients with AOSD. RESULTS: Significantly higher IFNgamma-producing Th cells and Th1/Th2 ratio in PB were found in patients with AOSD than in healthy controls. Percentages of IFNgamma-producing Th cells and Th1/Th2 ratio in PB correlated significantly with clinical activity score and serum IL18 levels in patients with AOSD. Increased ratio of Th1/Th2 cytokine transcripts was seen in the biopsy specimens of evanescent rash and synovitis from patients with AOSD compared with normal skin controls and patients with OA. Th cell cytokine pattern in PB and cytokine mRNA expression in synovium were similar for patients with AOSD and with RA. After 3 months' treatment, clinical remission was associated with a marked decrease in the percentages of cytokine-producing Th1 cells, but not of the Th2 cells. CONCLUSION: A predominance of Th1 cytokine may precipitate the pathogenesis of AOSD. PMID- 15361392 TI - Antiphospholipid syndrome associated with infections: clinical and microbiological characteristics of 100 patients. AB - OBJECTIVE: To describe and analyse the clinical characteristics of 100 patients with antiphospholipid syndrome (APS) associated with infections. METHODS: Patients were identified by a computer assisted search (Medline) of published reports to locate all cases of APS published in English, Spanish, and French from 1983 to 2003. The bilateral Fisher exact test was used for statistics. RESULTS: 59 female and 41 male patients were identified (mean (SD) age, 32 (18) years (range 1 to 78)): 68 had primary APS, 27 had systemic lupus erythematosus, two had "lupus-like" syndrome, two had inflammatory bowel disease, and one had rheumatoid arthritis. APS presented as a catastrophic syndrome in 40% of cases. The main clinical manifestations of APS included: pulmonary involvement (39%), skin involvement (36%), and renal involvement (35%; nine with renal thrombotic microangiopathy, RTMA). The main associated infections and agents included skin infection (18%), HIV (17%), pneumonia (14%), hepatitis C (13%), and urinary tract infection (10%). Anticoagulation was used in 74%, steroids in 53%, intravenous immunoglobulins in 20%, cyclophosphamide in 12%, plasma exchange in 12%, and dialysis in 9.6%. Twenty three patients died following infections and thrombotic episodes (16 with catastrophic APS). Patients given steroids had a better prognosis (p = 0.024). The presence of RTMA and requirement for dialysis carried a worse prognosis (p = 0.001 and p = 0.035, respectively). CONCLUSIONS: Various different infections can be associated with thrombotic events in patients with APS, including the potentially lethal subset termed catastrophic APS. Aggressive treatment with anticoagulation, steroids, and appropriate antibiotic cover is necessary to improve the prognosis. PMID- 15361393 TI - Autologous stem cell transplantation for refractory juvenile idiopathic arthritis: analysis of clinical effects, mortality, and transplant related morbidity. AB - OBJECTIVE: To evaluate the safety and efficacy of autologous stem cell transplantation (ASCT) for refractory juvenile idiopathic arthritis (JIA). DESIGN: Retrospective analysis of follow up data on 34 children with JIA who were treated with ASCT in nine different European transplant centres. Rheumatological evaluation employed a modified set of core criteria. Immunological reconstitution and infectious complications were monitored at three month intervals after transplantation. RESULTS: Clinical follow up ranged from 12 to 60 months. Eighteen of the 34 patients (53%) with a follow up of 12 to 60 months achieved complete drug-free remission. Seven of these patients had previously failed treatment with anti-TNF. Six of the 34 patients (18%) showed a partial response (ranging from 30% to 70% improvement) and seven (21%) were resistant to ASCT. Infectious complications were common. There were three cases of transplant related mortality (9%) and two of disease related mortality (6%). CONCLUSIONS: ASCT in severely ill patients with JIA induces a drug-free remission of the disease and a profound increase in general wellbeing in a substantial proportion of patients, but the procedure carries a significant mortality risk. The following adjustments are proposed for future protocols: (1) elimination of total body irradiation from the conditioning regimen; (2) prophylactic administration of antiviral drugs and intravenous immunoglobulins until there is a normal CD4+ T cell count. PMID- 15361394 TI - Immune activation in the small intestine in patients with rheumatoid arthritis. AB - OBJECTIVES: To determine whether inflammation in the gut associated immune system is activated in rheumatoid arthritis (RA). The expression of chemokine receptor- (CCR4, CCR5) and cytokine- (interleukin (IL)2, IL10, interferon gamma (IFNgamma), tumour necrosis factor alpha (TNFalpha), and transforming growth factor beta (TGFbeta)) specific mRNA in intestinal biopsy samples from patients with RA was examined. METHODS: Duodenal biopsy samples from 13 patients with RA and 15 control subjects were studied. The mRNA expression of CCR4, CCR5, IL2, IL10, IFNgamma, TNFalpha, and TGFbeta in intestinal biopsy samples was demonstrated by real time quantitative reverse transcriptase-polymerase chain reaction. RESULTS: The mRNA expression of CCR4, CCR5, and IL10 in intestinal biopsy samples was increased in patients with RA in comparison with control subjects (p = 0.001, p = 0.046, p = 0.019). No difference in the expression levels of IL2, IFNgamma, TNFalpha, or TGFbeta was seen between patients with RA and controls. CONCLUSIONS: The increased intestinal mRNA expression of IL10, CCR5, and CCR4 suggests that gut associated immune cells are activated in patients with RA. PMID- 15361395 TI - Hand cortical bone mass and its associations with radiographic joint damage and fractures in 50-70 year old female patients with rheumatoid arthritis: cross sectional Oslo-Truro-Amsterdam (OSTRA) collaborative study. AB - OBJECTIVE: To investigate the relationship between hand bone mineral density (BMD) and radiographic joint damage, and between hand BMD and fractures in 50-70 year old women with longstanding RA. METHODS: Demographic, clinical data, and imaging data on hand radiographs and Genants vertebral deformity score on spine radiographs were collected from 135 women with RA of > or =5 years, recruited from three European rheumatology clinics. Metacarpal hand BMD was measured by digital hand x ray radiogrammetry (DXR), and hip and lumbar spine BMD by dual x ray absorptiometry (DXA). Multiple regression analyses were used to examine associations between hand BMD and radiographic joint damage, and hand BMD and fractures. RESULTS: Hand BMD was strongly and independently associated with radiographic hand joint damage in a linear regression model adjusted for age, centre, BMI, disease duration, RF, 18 deformed joint count, ESR, and femoral neck BMD. In a multivariate logistic regression model adjusted for relevant variables, hand BMD and femoral neck BMD, but not spine BMD, were independently associated with vertebral deformities and with non-vertebral fractures. CONCLUSION: BMD measured by DXR on conventional hand radiographs in patients with RA may potentially be used as an indicator of joint damage and of vertebral and non vertebral fracture risk. PMID- 15361396 TI - Serum cytokine levels related to multiple dimensions of fatigue in patients with primary Sjogren's syndrome. AB - OBJECTIVE: To test whether serum levels of selected cytokines relate to different dimensions of fatigue in patients with primary Sjogren's syndrome (pSS). METHODS: Sixty female patients with pSS filled out a questionnaire to assess multiple dimensions of fatigue. Scores were compared with values in a population based control group (n = 139). Levels of interleukin (IL)1beta, IL2, IL6, IL10, and tumour necrosis factor alpha were measured in serum with commercial sandwich ELISAs. The relationship between self reported dimensions of fatigue and these serum cytokine levels was determined. RESULTS: Patients with pSS had high scores at all dimensions of fatigue (p<0.001): general fatigue, physical fatigue, reduced activity, reduced motivation, and mental fatigue. Fatigue levels were not related to serum cytokine levels. The incidental finding that reduced motivation was higher in patients with detectable serum levels of IL10 (p = 0.04) disappeared after correction for multiple testing. CONCLUSION: Fatigue is prominent in patients with pSS and involves all dimensions of fatigue. The findings do not suggest a widespread effect of circulating cytokines on multiple aspects of fatigue. PMID- 15361397 TI - Relationship between disease activity and infection in patients with spondyloarthropathies. AB - OBJECTIVE: To assess the relationship between disease activity and signs and symptoms of infection in Mexican patients with spondyloarthropathies (SpA). METHODS: A cross sectional study of 95 non-selected patients with SpA (62 men; mean age 26.4 years), who were examined for signs and symptoms of infection and their association with disease activity. 52 had ankylosing spondylitis (AS), 32 undifferentiated SpA (uSpA), 6 chronic reactive arthritis (ReA), and 5 psoriatic arthritis (PsA). Categorical data were analysed by chi(2) or Fisher's tests. RESULTS: 53 (56%) patients had infections: 41 (43%) upper respiratory tract (URT), 34 (36%) enteric, and 20 (21%) genitourinary infections. More infections occurred in HLA-B27 positive patients as a whole (39 v 5; p = 0.003) and in uSpA (12 v 2; p = 0.005). In AS and uSpA, infections occurred in approximately 50%. 30/39 (77%) patients with active disease (group A) and 23/56 (41%) (group B) (p = 0.001) had infection. There were more enteric infections in group A (47%; p<0.001) and more URT infections in group B (52%; p = NS). 22/30 (73%) patients attributed disease activity to infection. CONCLUSION: Enteric, and less commonly, URT infections in Mexican patients with SpA, particularly those who were HLA-B27 positive, seem to have a role in the active phase of AS and uSpA. PMID- 15361398 TI - Low dose warfarin treatment for calcinosis in patients with systemic sclerosis. AB - OBJECTIVE: To evaluate the effect of low doses of warfarin in patients with systemic sclerosis with disseminated subcutaneous calcinosis. METHODS: Three patients with disseminated subcutaneous calcinosis were treated with low doses of warfarin for 1 year. Subcutaneous calcinotic lesions, coagulation blood parameters, and the tendency for bleeding were followed up during the year. RESULTS: Two of the patients, who had newly diagnosed, diffuse, and relatively small calcinotic lesions, responded to warfarin treatment, with complete resolution of the calcinosis. The other patient, with larger and longer standing calcinotic lesions, did not respond to warfarin treatment. None of the three patients showed a prolongation of prothrombin time or partial thromboplastin time, nor did any have an increased tendency for bleeding. CONCLUSIONS: Low dose warfarin may serve as an effective treatment for calcinosis in a selected group of patients who have small and relatively new onset calcinosis. This treatment does not prolong the coagulation of blood and there is no increased tendency for bleeding. PMID- 15361399 TI - An unusual case of "giant cell arteritis". PMID- 15361400 TI - Visceral leishmaniasis resembling systemic lupus erythematosus. PMID- 15361401 TI - Neurovascular mechanisms as a possible cause of remission of rheumatoid arthritis in hemiparetic limbs. PMID- 15361402 TI - Severe gouty arthritis refractory to anti-inflammatory drugs: treatment with anti tumour necrosis factor alpha as a new therapeutic option. PMID- 15361403 TI - Two young girls with pyogenic sacroiliitis. PMID- 15361404 TI - Use of minocycline in rheumatoid arthritis: a district general hospital experience. PMID- 15361405 TI - Toxic myopathy induced by the ingestion of loquat leaf extract. PMID- 15361406 TI - Tumour necrosis factor receptor associated periodic syndrome (TRAPS) with central nervous system involvement. PMID- 15361407 TI - Effectiveness of adenotonsillectomy in children with mild symptoms of throat infections or adenotonsillar hypertrophy: open, randomised controlled trial. AB - OBJECTIVE: To assess the effectiveness of adenotonsillectomy in children with mild symptoms of throat infections or adenotonsillar hypertrophy. DESIGN: Open, randomised controlled trial. SETTING: 21 general hospitals and three academic centres in the Netherlands. PARTICIPANTS: 300 children aged 2-8 years requiring adenotonsillectomy. INTERVENTION: Adenotonsillectomy compared with watchful waiting. MAIN OUTCOME MEASURES: Episodes of fever, throat infections, upper respiratory tract infections, and health related quality of life. RESULTS: During the median follow up period of 22 months, children in the adenotonsillectomy group had 2.97 episodes of fever per person year compared with 3.18 in the watchful waiting group (difference -0.21, 95% confidence interval -0.54 to 0.12), 0.56 throat infections per person year compared with 0.77 (-0.21, -0.36 to 0.06), and 5.47 upper respiratory tract infections per person year compared with 6.00 (-0.53, -0.97 to -0.08). No clinically relevant differences were found for health related quality of life. Adenotonsillectomy was more effective in children with a history of three to six throat infections than in those with none to two. 12 children had complications related to surgery. CONCLUSION: Adenotonsillectomy has no major clinical benefits over watchful waiting in children with mild symptoms of throat infections or adenotonsillar hypertrophy. PMID- 15361408 TI - The hydrodynamic radii of macromolecules and their effect on red blood cell aggregation. AB - The effects of nonionic polymers on human red blood cell (RBC) aggregation were investigated. The hydrodynamic radius (Rh) of individual samples of dextran, polyvinylpyrrolidone, and polyoxyethylene over a range of molecular weights (1,500-2,000,000) were calculated from their intrinsic viscosities using the Einstein viscosity relation and directly measured by quasi-elastic light scattering, and the effect of each polymer sample on RBC aggregation was studied by nephelometry and low-shear viscometry. For all three polymers, despite their different structures, samples with Rh <4 nm were found to inhibit aggregation, whereas those with Rh >4 nm enhanced aggregation. Inhibition increased with Rh and was maximal at approximately 3 nm; above 4 nm the pro-aggregant effect increased with Rh. For comparison, the Rh of 12 plasma proteins were calculated from literature values of intrinsic viscosity or diffusion coefficient. Each protein known to promote RBC aggregation had Rh >4 nm, whereas those with Rh <4 nm either inhibited or had no effect on aggregation. These results suggest that the influence of a nonionic polymer or plasma protein on RBC aggregation is simply a consequence of its size in an aqueous environment, and that the specific type of macromolecule is of minor importance. PMID- 15361409 TI - Electrostatic mechanisms underlie neomycin block of the cardiac ryanodine receptor channel (RyR2). AB - Neomycin is a large, positively charged, aminoglycoside antibiotic that has previously been shown to induce a voltage-dependent substate block in the cardiac isoform of the ryanodine receptor (RyR2). It was proposed that block involved an electrostatic interaction between neomycin and putative regions of negative charge in both the cytosolic and luminal mouths of the pore. In this study, we have attempted to screen charge by increasing potassium concentration in single channel experiments. Neomycin block is apparent at both cytosolic and luminal faces of the channel in all K+ concentrations tested and alterations in K+ concentration have no effect on the amplitudes of the neomycin-induced substates. However, the kinetics of both cytosolic and luminal block are sensitive to changes in K+ concentration. In both cases increasing the K+ concentration leads to an increase in dissociation constant (KD). Underlying these changes are marked increases in rates of dissociation (k(off)), with little change in rates of association (k(on)). The increase in k(off) is more marked at the luminal face of the channel. Changes in K+ concentration also result in alterations in the voltage dependence of block. We have interpreted these data as supporting the proposal that neomycin block of RyR2 involves electrostatic interactions with the polycation forming a poorly fitting "plug" in the mouths of the conduction pathway. These observations emphasize the usefulness of neomycin as a probe for regions of charge in both the cytosolic and luminal mouths of the RyR2 pore. PMID- 15361410 TI - CFTR: a cysteine at position 338 in TM6 senses a positive electrostatic potential in the pore. AB - We investigated the accessibility to protons and thiol-directed reagents of a cysteine substituted at position 338 in transmembrane segment 6 (TM6) of CFTR to test the hypothesis that T338 resides in the pore. Xenopus oocytes expressing T338C CFTR exhibited pH-dependent changes in gCl and I-V shape that were specific to the substituted cysteine. The apparent pKa of T338C CFTR was more acidic than that expected for a cysteine or similar simple thiols in aqueous solution. The pKa was shifted toward alkaline values when a nearby positive charge (R334) was substituted with neutral or negatively charged residues, consistent with the predicted influence of the positive charge of R334, and perhaps other residues, on the titration of a cysteine at 338. The relative rates of chemical modification of T338C CFTR by MTSET+ and MTSES- were also altered by the charge at 334. These observations support a model for CFTR that places T338 within the anion conduction path. The apparent pKa of a cysteine substituted at 338 and the relative rates of reaction of charged thiol-directed reagents provide a crude measure of a positive electrostatic potential that may be due to R334 and other residues near this position in the pore. PMID- 15361411 TI - Rearrangement of the 16S precursor subunits is essential for the formation of the active 20S proteasome. AB - Proteasome-dependent proteolysis is essential for a number of key cellular processes and requires a sophisticated biogenesis pathway to function. Here, we have arrested the assembly process in its dynamic progression at the short-lived 16S state. Structural analysis of the 16S proteasome precursor intermediates by electron microscopy, and single particle analysis reveals major conformational changes in the structure of the beta-ring in comparison with one-half of the 20S proteasome. The individual beta-subunits in the 16S precursor complex rotate with respect to their positions in the x-ray crystallographic structure of the fully assembled 20S. This rearrangement results in a movement of the catalytic residue threonine-1 from the protected location in 16S precursor complexes to a more exposed position in the 20S structure. Thereby, our findings provide a molecular explanation for the structural rearrangements necessary for the dimerization of two 16S precursor complexes and the subsequent final maturation to active 20S proteasomes. PMID- 15361412 TI - Rheological analysis and measurement of neutrophil indentation. AB - Aspects of neutrophil mechanical behavior relevant to the formation of adhesive contacts were assessed by measuring the dependence of the contact area between the cell and a spherical substrate under controlled loading. Micropipettes were used to bring neutrophils into contact with spherical beads under known forces, and the corresponding contact area was measured over time. The neutrophil was modeled as a viscous liquid drop with a constant cortical tension. Both the equilibrium state and the dynamics of the approach to equilibrium were examined. The equilibrium contact area increased monotonically with force in a manner consistent with a cell cortical tension of 16-24 pN/microm. The dynamic response matched predictions based on a model of the cell as a growing drop using published values for the effective viscosity of the cell. The contact pressure between the cell and substrate at equilibrium is predicted to depend on the curvature of the contacting substrate, but to be independent of the impingement force. The approach to equilibrium was rapid, such that the time-averaged stress for a two-second impingement was within 20% of the equilibrium value. These results have implications for the role of mechanical force in the formation of adhesive contacts. PMID- 15361413 TI - Neutrophil adhesive contact dependence on impingement force. AB - Neutrophil capture and recruitment from the circulation requires the formation of specific receptor/ligand bonds under hydrodynamic forces. In the present study we examine bond formation between beta2-integrins on neutrophils and immobilized ICAM-1 while using micropipettes to control the force of contact between the cell and substrate. Magnesium was used to induce the high affinity conformation of the integrins, and bond formation was assessed by measuring the probability of adhesion during repeated contacts. Increasing the impingement force caused an increase in the contact area and led to a proportional increase in adhesion probability (from approximately 20 to 50%) over the range of forces tested (50 350 pN). In addition, different-sized beads were used to change the force per unit area in the contact zone (contact stress). We conclude that for a given contact stress, the rate of bond formation increases linearly with contact area, but that increasing contact stress results in higher intrinsic rates of bond formation. PMID- 15361414 TI - Reversible mechanical unfolding of single ubiquitin molecules. AB - Single-molecule manipulation techniques have enabled the characterization of the unfolding and refolding process of individual protein molecules, using mechanical forces to initiate the unfolding transition. Experimental and computational results following this approach have shed new light on the mechanisms of the mechanical functions of proteins involved in several cellular processes, as well as revealed new information on the protein folding/unfolding free-energy landscapes. To investigate how protein molecules of different folds respond to a stretching force, and to elucidate the effects of solution conditions on the mechanical stability of a protein, we synthesized polymers of the protein ubiquitin and characterized the force-induced unfolding and refolding of individual ubiquitin molecules using an atomic-force-microscope-based single molecule manipulation technique. The ubiquitin molecule was highly resistant to a stretching force, and the mechanical unfolding process was reversible. A model calculation based on the hydrogen-bonding pattern in the native structure was performed to explain the origin of this high mechanical stability. Furthermore, pH effects were studied and it was found that the forces required to unfold the protein remained constant within a pH range around the neutral value, and forces decreased as the solution pH was lowered to more acidic values. PMID- 15361415 TI - Calculated coupling of transmembrane electron and proton transfer in dihemic quinol:fumarate reductase. AB - The quinol:fumarate reductase of Wolinella succinogenes binds a low- and a high potential heme b group in its transmembrane subunit C. Both hemes are part of the electron transport chain between the two catalytic sites of this redox enzyme. The oxidation-reduction midpoint potentials of the hemes are well established but their assignment in the structure has not yet been determined. By simulating redox titrations, using continuum electrostatics calculations, it was possible to achieve an unequivocal assignment of the low- and high-potential hemes to the distal and proximal positions in the structure, respectively. Prominent features governing the differences in midpoint potential between the two hemes are the higher loss of reaction field energy for the proximal heme and the stronger destabilization of the oxidized form of the proximal heme due to several buried Arg and Lys residues. According to the so-called "E-pathway hypothesis", quinol:fumarate reductase has previously been postulated to exhibit a novel coupling of transmembrane electron and proton transfer. Simulation of heme b reduction indicates that the protonation state of the conserved residue Glu C180, predicted to play a key role in this process, indeed depends on the redox state of the hemes. This result clearly supports the E-pathway hypothesis. PMID- 15361416 TI - Predictors of survival in sporadic Creutzfeldt-Jakob disease and other human transmissible spongiform encephalopathies. AB - A collaborative study of human transmissible spongiform encephalopathies has been carried out from 1993 to 2000 and includes data from 10 national registries, the majority in Western Europe. In this study, we present analyses of predictors of survival in sporadic (n = 2304), iatrogenic (n = 106) and variant Creutzfeldt Jakob disease (n = 86) and in cases associated with mutations of the prion protein gene (n = 278), including Gerstmann-Straussler-Scheinker syndrome (n = 24) and fatal familial insomnia (n = 41). Overall survival for each disease type was assessed by the Kaplan-Meier method and the multivariate analyses by the Cox proportional hazards model. In sporadic disease, longer survival was correlated with younger age at onset of illness, female gender, codon 129 heterozygosity, presence of CSF 14-3-3 protein and type 2a prion protein type. The ability to predict survival based on patient covariates is important for diagnosis and counselling, and the characterization of the survival distributions, in the absence of therapy, will be an important starting point for the assessment of potential therapeutic agents in the future. PMID- 15361417 TI - FDG-PET improves tumour detection in patients with paraneoplastic neurological syndromes. AB - To determine the usefulness of [18F]fluorodeoxyglucose (FDG) whole body FDG-PET in the diagnosis of tumours in patients with paraneoplastic neurological syndromes (PNS), we prospectively studied 20 patients with paraneoplastic antibodies in whom conventional imaging gave negative or inconclusive results for the presence of tumour. All 20 patients had neurological manifestations compatible with PNS and well-characterized paraneoplastic antibodies (12 anti-Hu, one anti-Hu and anti-CV2, one anti-CV2, four anti-Yo, one anti-Ri and one anti amphiphysin). The mean delay between the onset of neurological symptoms and FDG PET was 10 months (range 1-54). In these 20 patients, abnormal uptake was demonstrated in 18 patients, with some patients having abnormal signal in several areas. We observed abnormal uptake in the mediastinum (13 cases), lung (two cases), breast (two cases), parotid gland (one case), or the cervical, supraclavicular or axillary lymph nodes (seven cases). Following FDG-PET, the histological diagnosis of the tumour was made in 14 patients (small cell lung carcinoma in eight cases, breast adenocarcinoma in two, lung adenocarcinoma in two, axillary metastasis of ovary carcinoma in one, and malignant thymoma in one). Two other patients with abnormal FDG uptake showed radiological evidence of lung cancer, but a histological diagnosis could not be obtained. In two other patients, initial FDG-PET showed abnormal FDG uptake that was not confirmed a few months later by repeat FDG-PET. In the two patients with negative FDG-PET, peritoneal carcinomatosis was diagnosed in one and no tumour was found in the other. In our series, the sensitivity of FDG-PET for tumour detection was >83% demonstrating a clear role of this technique in the management of patients with PNS. However, in our series, the specificity of FDG uptake was only 25% due to unexplained abnormal FDG uptake in three patients and in abnormal FDG uptake due to a benign tumour in one patient. Over the study period, we saw 73 other patients with PNS and paraneoplastic antibodies. A tumour was demonstrated in 71 out of 73 by conventional techniques. Since false-positive and false-negative results are possible with FDG-PET and in most patients with PNS, the tumour is demonstrated by conventional techniques, we believe that FDG-PET should be reserved, at the moment, for patients with well-defined PNS antibodies when conventional imaging fails to identify a tumour or when lesions are difficult to biopsy. PMID- 15361418 TI - Using monoamine oxidase type B inhibitors in Parkinson's disease. PMID- 15361420 TI - Ethnic profile of the doctors in the United Kingdom. PMID- 15361421 TI - Doctors and nurses with HIV and AIDS in sub-Saharan Africa. PMID- 15361422 TI - Early life intelligence and adult health. PMID- 15361424 TI - Nationwide protocol is needed for investigating baby deaths. PMID- 15361435 TI - Doctors accused of discriminating against Down's syndrome patients. PMID- 15361437 TI - Seven doctors to face GMC over Shipman inquiry findings. PMID- 15361441 TI - Slugging it out over health care, stem cells, and abortion. PMID- 15361442 TI - Couple allowed to select an embryo to save sibling. PMID- 15361443 TI - Continuing violence in Sudan prompts calls for sanctions. PMID- 15361444 TI - Can care management enhance integration of primary and specialty care? PMID- 15361445 TI - Using computers for assessment in medicine. PMID- 15361447 TI - The journey: Parkinson's disease. PMID- 15361448 TI - Gender related disorders. PMID- 15361449 TI - Ethics of screening for asymptomatic herpes virus type 2 infection. PMID- 15361450 TI - Research bureaucracy in the United Kingdom: seeking a balance: response from the Department of Health and COREC. PMID- 15361451 TI - Research bureaucracy in the United Kingdom: good governance is needed. PMID- 15361452 TI - Research bureaucracy in the United Kingdom: ethics committees have important roles in research. PMID- 15361453 TI - Research bureaucracy in the United Kingdom: research governance is about protection, not convenience. PMID- 15361454 TI - Research bureaucracy in the United Kingdom: time has come to face research governance. PMID- 15361455 TI - Editorial independence at the BMJ:.and the author apologises unreservedly. PMID- 15361456 TI - Editorial independence at the BMJ: ... JAMA editor expresses disappointment at letter's smear... PMID- 15361457 TI - Editorial independence at the BMJ: NEJM editor says facts in letter are wrong and accusations reckless... PMID- 15361458 TI - Research bureaucracy in the United Kingdom: practical example illustrates problems of ethical review of genetics. PMID- 15361459 TI - Research bureaucracy in the United Kingdom: ask for help. PMID- 15361460 TI - Predicting bacterial cause in infectious conjunctivitis: Chlamydia needs to be taken into account. PMID- 15361461 TI - Predicting bacterial cause in infectious conjunctivitis: "history of conjunctivitis" needs clarifying. PMID- 15361462 TI - Predicting bacterial cause in infectious conjunctivitis: three more questions for "conjunctivitis" may be important. PMID- 15361463 TI - Predicting bacterial cause in infectious conjunctivitis: why say itching counts against bacterial infection in conjunctivitis? PMID- 15361464 TI - All clinical trials must be reported in detail and made publicly available. PMID- 15361465 TI - Hospital chaplaincy units show bias towards Christianity. PMID- 15361471 TI - Galectin-3/AGE-receptor 3 knockout mice show accelerated AGE-induced glomerular injury: evidence for a protective role of galectin-3 as an AGE receptor. AB - We previously showed that mice lacking galectin-3/AGE-receptor 3 develop accelerated diabetic glomerulopathy. To further investigate the role of galectin 3/AGE-receptor function in the pathogenesis of diabetic renal disease, galectin-3 knockout (KO) and coeval wild-type (WT) mice were injected for 3 months with 30 microg/day of N(epsilon)-carboxymethyllysine (CML)-modified or unmodified mouse serum albumin (MSA). Despite receiving equal doses of CML, KO had higher circulating and renal AGE levels and showed more marked renal functional and structural changes than WT mice, with significantly higher proteinuria, albuminuria, glomerular, and mesangial area and glomerular sclerosis index. Renal 4-hydroxy-2-nonenal content and NFkappaB activation were also more pronounced in KO-CML vs. WT-CML. Kidney mRNA levels of fibronectin, laminin, collagen IV, and TGF-beta were up-regulated, whereas those of matrix metalloproteinase-2 and -14 were down-regulated, again more markedly in KO-CML than WT-CML mice. Basal and CML-induced RAGE and 80K-H mRNA levels were higher in KO vs. WT mice. MSA injection did not produce any significant effect in both genotypes. The association of galectin-3 ablation with enhanced susceptibility to AGE-induced renal disease, increased AGE levels and signaling, and altered AGE-receptor pattern indicates that galectin-3 is operating in vivo as an AGE receptor to afford protection toward AGE-dependent tissue injury. PMID- 15361472 TI - Vascular endothelial growth factor-C gene therapy restores lymphatic flow across incision wounds. AB - Edema and insufficient blood perfusion are common problems in reconstructive surgery. The blood vasculature is reconstructed in microvascular flaps, whereas lymphatic vessel function is lost after surgical incision. Here, we demonstrate that vascular endothelial growth factor C (VEGF-C) gene transfer can be used to reconstruct a lymphatic vessel network severed by incision of skin flaps. We used adenoviral VEGF-C gene transfer at the edges of epigastric skin flaps in mice. Our results show that VEGF-C gene expression results in the formation of anastomoses between the lymphatic vessels of the skin flap and the surrounding lymphatic vasculature. Some spontaneous lymphangiogenesis also took place in the control mice, but the lymphatic vessels generated remained nonfunctional even 2 months postoperatively. In contrast, the VEGF-C treated mice demonstrated persistent lymphatic vessel function during the 2 month follow-up despite the transient nature of the adenoviral VEGF-C gene expression. The restoration of lymphatic function by VEGF-C in skin flaps provides new tools to promote vascular perfusion and to reduce tissue edema in skin and muscle flaps. These results have important implications for the prevention and treatment of surgically induced secondary lymphedema. PMID- 15361473 TI - Editorial I: Guidelines for routine preoperative testing. PMID- 15361474 TI - Editorial II: Who is at increased risk of pulmonary aspiration? PMID- 15361475 TI - Editorial III: Anaphylaxis and anaesthesia--all clear now? PMID- 15361476 TI - Articaine for sub-Tenon's and peribulbar anaesthesia in cataract surgery. PMID- 15361477 TI - Does bispectral analysis add anything but complexity? BIS sub-components may be superior to BIS for detection of awareness. PMID- 15361478 TI - ETT impingement during railroading over the FOB. PMID- 15361479 TI - Novel approach to management of a posterior tracheal tear complicating percutaneous tracheostomy. PMID- 15361480 TI - Ultrasound imaging of the axillary vein--anatomical basis for central venous access. PMID- 15361481 TI - Food allergy in irritable bowel syndrome: new facts and old fallacies. AB - The notion of food allergy in irritable bowel syndrome (IBS) is not new. However, recent evidence suggests significant reduction in IBS symptom severity in patients on elimination diets, provided that dietary elimination is based on foods against which the individual had raised IgG antibodies. These findings should encourage studies dissecting the mechanisms responsible for IgG production against dietary antigens and their putative role in IBS PMID- 15361482 TI - The role of Cdx genes in the mammalian gut. PMID- 15361483 TI - Is there a SERT-ain association with IBS? PMID- 15361484 TI - Diet and relapsing ulcerative colitis: take off the meat? PMID- 15361485 TI - The Munich Barrett follow up study: suspicion of Barrett's oesophagus based on either endoscopy or histology only--what is the clinical significance? AB - BACKGROUND: The incidence of distal oesophageal adenocarcinoma is rising, with chronic reflux and Barrett's oesophagus being considered risk factors. Reliable detection of Barrett's oesophagus during upper endoscopy is therefore mandatory but requires both endoscopy and histology for confirmation. Appropriate management of patients with endoscopic suspicion but negative on histology, or vice versa, or of patients with no endoscopic suspicion but with a biopsy diagnosis of intestinal metaplasia at the gastro-oesophageal junction, has not yet been studied prospectively. PATIENTS AND METHODS: In a prospective multicentre study, 929 patients (51% male, mean age 50 years) referred for upper gastrointestinal endoscopy were included; 59% had reflux symptoms. The endoscopic aspect of the Z line and any suspicion of Barrett's oesophagus were noted, and biopsies were taken in all patients from the Z line (n = 4), gastric cardia (n = 2), and body and antrum (n = 2 each). Biopsies positive for specialised intestinal metaplasia (SIM) were reviewed by a reference pathologist for a final Barrett's oesophagus diagnosis. All patients with endoscopic and/or histological suspicion of Barrett's oesophagus were invited for a follow up endoscopy; the remaining cases (no endoscopic or histological suspicion of Barrett's oesophagus) were followed clinically. RESULTS: Of 235 patients positive for Barrett's oesophagus on endoscopy and/or histology, 63% agreed to undergo repeat endoscopy (mean follow up period 30.5 months). 46% of patients with an endoscopic Barrett's oesophagus diagnosis but no histological confirmation (group A) showed the same distribution, a further 42% did not have Barrett's oesophagus, and 11% had confirmed Barrett's oesophagus on both endoscopy and biopsy on follow up. In the group with a histological Barrett's oesophagus diagnosis but negative on initial endoscopy (group B), follow up showed the same in 26% whereas 46% had no Barrett's oesophagus, and confirmed Barrett's oesophagus (endoscopy plus histology) was diagnosed in 17%. Of the study population, 16 patients had Barrett's oesophagus on initial endoscopy confirmed by histology which remained constant in 70% at follow up (group C). Of the remaining patients without an initial Barrett's oesophagus diagnosis on either endoscopy or histology (group D) and only clinical follow up (mean follow up period 38 months), one confirmed Barrett's oesophagus case was found among 100 patients re-endoscoped outside of the study protocol. However, no single case of dysplasia or cancer of the distal oesophagus was detected in any patient during the study period. CONCLUSIONS: Even in a specialised gastroenterology setting, reproducibility of presumptive endoscopic or histological diagnoses of Barrett's oesophagus at follow up were poor. Only 10-20% of cases with either endoscopic or histological suspicion of Barrett's oesophagus had established Barrett's oesophagus after 2.5 years of follow up. The risk of dysplasia in this population was very low and hence meticulous follow up may not be required. PMID- 15361486 TI - Trefoil factor 1 is required for the commitment programme of mouse oxyntic epithelial progenitors. AB - BACKGROUND: Trefoil factor 1 (TFF1/pS2) is a major secretory product of the stomach and TFF1 knockout mice constantly develop adenomas and occasional carcinomas in the pyloric antrum. AIM: To analyse the role of TFF1 in the differentiation of gastric epithelial cell lineages using oxyntic mucosae from normal and TFF1 knockout mice. METHODS: The various cell lineages were labelled using specific markers of pit, neck, parietal, and enteroendocrine cells. Patterns of TFF1, TFF2, and TFF3 expressions were defined using western blotting, immunohistochemistry, and/or immunogold electron microscopy. RESULTS: In normal mice, starting from postnatal day 1 (P1), TFF1 and TFF2 were produced by mucus secreting cells of the developing epithelium. At P7, TFF3 expression occurred in pit and parietal cells. When oxyntic glands were compartmentalised, at P21 and in older mice, TFF1 and TFF2 were expressed in pit and neck cells, respectively, and TFF3 was no longer in parietal cells but became a feature of zymogenic cells. In TFF1 deficient mice, alteration of oxyntic epithelial differentiation became obvious at P21, showing significant amplification of pit cells at the expense of parietal cells. At the molecular level, lack of TFF1 induced dramatic inhibition of TFF2 expression and more precocious TFF3 expression. CONCLUSION: In the oxyntic mucosa, all three TFFs are produced in a lineage specific manner and TFF1 is essential in maintaining the normal commitment programme of epithelial progenitors. PMID- 15361487 TI - Cdx1 induced intestinal metaplasia in the transgenic mouse stomach: comparative study with Cdx2 transgenic mice. AB - BACKGROUND AND AIMS: Gastric intestinal metaplasia, which is mainly induced by Helicobacter pylori infection, is thought to be a precancerous lesion of gastric adenocarcinoma. Intestinal metaplastic mucosa expresses intestine specific homeobox genes, Cdx1 and Cdx2, in the human gastric mucosa. We and others have reported that ectopic expression of Cdx2 in the gastric epithelium generates intestinal metaplasia in the transgenic mouse model. METHODS: To clarify the differences in the roles of Cdx1 and Cdx2 in intestinal metaplasia, we generated transgenic mice expressing Cdx1 in the gastric mucosa and compared Cdx1 induced gastric mucosal morphological changes with Cdx2 induced intestinal metaplasia. RESULTS: The gastric mucosa in Cdx1 transgenic mice was completely replaced by intestinal metaplastic mucosa, consisting of all four intestinal epithelial cell types: absorptive enterocytes, goblet, enteroendocrine, and Paneth cells. Paneth cells, which were not recognised in Cdx2 transgenic mice, were in the upper portion of the intestinal metaplastic mucosa. Pseudopyloric gland metaplasia, which was induced in Cdx2 transgenic mice, was not recognised in Cdx1 transgenic mice. Proliferating cell nuclear antigen (PCNA) positive cells were diffusely scattered in Cdx1 induced intestinal metaplastic mucosa while PCNA positive cells in Cdx2 induced intestinal metaplastic mucosa were in the base of the metaplastic mucosa. Intestinal metaplastic mucosa of Cdx1 transgenic mouse stomach was significantly thicker than that of wild-type or Cdx2 transgenic mouse stomach. CONCLUSIONS: We have confirmed that Cdx1 induced gastric intestinal metaplasia but that it differed from Cdx2 induced intestinal metaplasia in differentiation, structure, and proliferation. PMID- 15361488 TI - Early interactions of Salmonella enterica serovar typhimurium with human small intestinal epithelial explants. AB - BACKGROUND: Salmonella enterica serovar typhimurium (S typhimurium) causes invasive gastroenteritis in humans, a disease involving significant penetration of the intestinal mucosa. However, few studies have been undertaken to investigate this interaction directly using differentiated human gut tissue. AIMS: To investigate the early interactions of an enteropathogenic strain of S typhimurium with human intestinal mucosa using human intestinal in vitro organ culture (IVOC). METHODS: Wild-type and mutant derivatives of S typhimurium TML were used to compare interactions with cultured human epithelial cells, bovine ligated loops, and human intestinal IVOC. RESULTS: S typhimurium TML was shown to attach to cultured Caco-2 brush border expressing cells and cause tissue damage and fluid accumulation in a ligated bovine loop model.S typhimurium TML bound predominantly to the mucus layer of human IVOC explants during the first four hours of IVOC incubation. From four to eight hours of IVOC incubation, small but characteristic foci of attaching and invading S typhimurium TML were detected as clusters of bacteria interacting with enterocytes, although there was no evidence for large scale invasion of explant tissues. Ruffling of enterocyte membranes associated with adherent Salmonella was visualised using electron microscopy. CONCLUSIONS: Human IVOC can be used as an alternative model for monitoring the interactions between S typhimurium and human intestinal epithelium, thus potentially offering insight into the early stages of human Salmonella induced gastroenteritis. PMID- 15361489 TI - A rare cause of gastrointestinal bleeding in a patient with hyper-IgE syndrome. PMID- 15361490 TI - Cellular detection of sst2A receptors in human gastrointestinal tissue. AB - BACKGROUND AND AIMS: Many neuroendocrine gastrointestinal tumours express receptors for the regulatory peptide somatostatin. Among the five existing somatostatin receptor (sst) subtypes, sst2A is the most frequently expressed in these tumours. However, little information is available about the cellular location of sst2A in corresponding non-neoplastic epithelial tissues. METHODS: We searched for sst2A immunoreactive cells in non-neoplastic gastrointestinal tissues, and evaluated their number and immunohistochemical characteristics with neuroendocrine markers. RESULTS: The gastric antrum showed numerous sst2A cells, situated in the epithelium, corresponding to gastrin containing neuroendocrine cells, while the gastric corpus was largely devoid of sst2A cells, including enterochromaffin-like cells. The remaining foregut, namely the duodenum and proximal jejunum, also contained a large number of sst2A cells, all being neuroendocrine cells and many of them characterised as gastrin cells. Sst2A cells were also detected in the midgut, in low numbers in the epithelium of the distal jejunum and ileum, but not in the appendix vermiformis, the caecum, or the hindgut, despite the large number of neuroendocrine cells present in this area. In addition, sst2A cells were found in the whole gastrointestinal tract in the myenteric and submucosal plexus. CONCLUSIONS: While sst2A receptors on antral gastrin cells presumably mediate somatostatin inhibition of gastrin secretion, the effects of somatostatin on motility and ion transport in the lower gastrointestinal tract may be mediated by sst2A receptors in the neural plexus. These data provide a molecular basis for the physiological actions of somatostatin in human gastrointestinal tissue. PMID- 15361491 TI - Enteropathy precedes type 1 diabetes in the BB rat. AB - BACKGROUND AND AIMS: There is increasing evidence implicating intestinal immune responses to dietary proteins in the pathogenesis of type 1 autoimmune diabetes (T1D). Here we investigated the association between intestinal pathology and dietary factors in T1D by examining the mucosal architecture in the BB rat model. METHODS: BB control (BBc) and diabetes prone (BBdp) rats were fed either a diabetes retardant hydrolysed casein based diet or one of two cereal based diets that promote the development of diabetes. Intestinal architecture was assessed in the jejunum by microdissection, histology, and immunohistology, and by measuring peroxidase activity and brush border invertase levels. RESULTS: Enteropathy was present in BBdp rats soon after weaning, as assessed by increases in crypt length and in the proliferative activity of crypt epithelial cells in the jejunum, and this remained constant until 120 days of age. There was also a decrease in invertase activity, as well as increased numbers of intraepithelial lymphocytes, increased levels of mucosal peroxidase activity, and infiltration of the mucosa by CD4(+) T lymphocytes. Equivalent enteropathy was present at all times in BBdp rats and was not influenced by the nature of the diet or by thymectomy at three weeks at age, procedures which prevent the development of diabetes. CONCLUSION: Enteropathy is a consistent feature in the diabetes prone BB rat but it precedes the onset of insulitis and appears to be due to mechanisms distinct from those which cause diabetes. The beneficial effects of the diabetes retardant hydrolysed casein diet on diabetes are not due to an effect on intestinal architecture per se but mucosal damage may be necessary for the development of autoreactivity in the pancreas. PMID- 15361492 TI - A waxing and waning skin rash. PMID- 15361493 TI - Symptom patterns in functional dyspepsia and irritable bowel syndrome: relationship to disturbances in gastric emptying and response to a nutrient challenge in consulters and non-consulters. AB - BACKGROUND: Our aim was to assess the relationship between gastric motor and sensory function and symptom patterns in community subjects and patients with functional dyspepsia (FD) or irritable bowel syndrome (IBS). METHODS: We recruited 291 asymptomatic blood donors, 151 symptomatic blood donors (recurrent abdominal pain or discomfort), and 40 patients with FD or IBS. Abdominal symptoms were assessed using the bowel disease questionnaire (BDQ) and, in addition, the most bothersome symptom complex identified (dysmotility-type, ulcer-type dyspepsia, or IBS). Gastric emptying time (GET (t(1/2), min)) was measured by (13)C-octanoic breath test and a nutrient challenge performed. Twenty randomly selected asymptomatic blood donors, 48 symptomatic blood donors (30 FD, 18 IBS), and 40 patients (23 FD, 17 IBS) had additional function testing. RESULTS: GET (t(1/2)) was significantly (p<0.05) longer in blood donors with FD symptoms (99 (6) min) and FD patients (110 (12) min) compared with asymptomatic controls (76.7 (7) min), but was not significant in IBS blood donors or patients. Overall, 25 of 48 blood donors with symptoms and 18 of 40 patients had slow gastric emptying. GET was most delayed in subjects with predominantly dysmotility-type symptoms (167 (36) min v controls; p<0.01). Symptom intensities after a nutrient challenge were significantly higher in FD patients and symptomatic blood donors compared with asymptomatic controls; 14 of 48 blood donors with symptoms and 16 of 40 patients had a symptom response to the nutrient challenge exceeding the response (mean (2SD)) of healthy asymptomatic controls. CONCLUSION: Gastric emptying and the global symptom response to a standardised nutrient challenge are abnormal in population based (non-health care seeking) subjects with dyspepsia. PMID- 15361494 TI - Association between a functional polymorphism in the serotonin transporter gene and diarrhoea predominant irritable bowel syndrome in women. AB - BACKGROUND AND AIMS: Serotonin (5-hydroxtryptamine, 5-HT) is an important factor in gut function, playing key roles in intestinal peristalsis and secretion, and in sensory signalling in the brain-gut axis. Removal from its sites of action is mediated by a specific protein called the serotonin reuptake transporter (SERT or 5-HTT). Polymorphisms in the promoter region of the SERT gene have effects on transcriptional activity, resulting in altered 5-HT reuptake efficiency. It has been speculated that such functional polymorphisms may underlie disturbance in gut function in individuals suffering with disorders such as irritable bowel syndrome (IBS). The aim of this study was to assess the potential association between SERT polymorphisms and the diarrhoea predominant IBS (dIBS) phenotype. SUBJECTS: A total of 194 North American Caucasian female dIBS patients and 448 female Caucasian controls were subjected to genotyping. METHODS: Leucocyte DNA of all subjects was analysed by polymerase chain reaction based technologies for nine SERT polymorphisms, including the insertion/deletion polymorphism in the promoter (SERT-P) and the variable tandem repeat in intron 2. Statistical analysis was performed to assess association of any SERT polymorphism allele with the dIBS phenotype. RESULTS: A strong genotypic association was observed between the SERT-P deletion/deletion genotype and the dIBS phenotype (p = 3.07x10(-5); n = 194). None of the other polymorphisms analysed was significantly associated with the presence of disease. CONCLUSIONS: Significant association was observed between dIBS and the SERT-P deletion/deletion genotype, suggesting that the serotonin transporter is a potential candidate gene for dIBS in women. PMID- 15361495 TI - Food elimination based on IgG antibodies in irritable bowel syndrome: a randomised controlled trial. AB - BACKGROUND: Patients with irritable bowel syndrome (IBS) often feel they have some form of dietary intolerance and frequently try exclusion diets. Tests attempting to predict food sensitivity in IBS have been disappointing but none has utilised IgG antibodies. AIMS: To assess the therapeutic potential of dietary elimination based on the presence of IgG antibodies to food. PATIENTS: A total of 150 outpatients with IBS were randomised to receive, for three months, either a diet excluding all foods to which they had raised IgG antibodies (enzyme linked immunosorbant assay test) or a sham diet excluding the same number of foods but not those to which they had antibodies. METHODS: Primary outcome measures were change in IBS symptom severity and global rating scores. Non-colonic symptomatology, quality of life, and anxiety/depression were secondary outcomes. Intention to treat analysis was undertaken using a generalised linear model. RESULTS: After 12 weeks, the true diet resulted in a 10% greater reduction in symptom score than the sham diet (mean difference 39 (95% confidence intervals (CI) 5-72); p = 0.024) with this value increasing to 26% in fully compliant patients (difference 98 (95% CI 52-144); p<0.001). Global rating also significantly improved in the true diet group as a whole (p = 0.048, NNT = 9) and even more in compliant patients (p = 0.006, NNT = 2.5). All other outcomes showed trends favouring the true diet. Relaxing the diet led to a 24% greater deterioration in symptoms in those on the true diet (difference 52 (95% CI 18 88); p = 0.003). CONCLUSION: Food elimination based on IgG antibodies may be effective in reducing IBS symptoms and is worthy of further biomedical research. PMID- 15361496 TI - Alteration of the spinal modulation of nociceptive processing in patients with irritable bowel syndrome. AB - BACKGROUND: Visceral hypersensitivity has been evidenced in patients with irritable bowel syndrome (IBS) but its mechanisms remain poorly elucidated. We investigated the spinal transmission of nociceptive signals in IBS patients by analysing the effects of rectal distensions on electromyographic recordings of the somatic nociceptive flexion (RIII) reflex, an objective index of spinal nociceptive processes. METHODS: Fourteen IBS and 10 healthy volunteers were included in the study. Slow ramp (40 ml/min) and rapid phasic (900 ml/min, 10, 20, 30, and 40 mm Hg) rectal distensions were randomly performed while the RIII reflex evoked by electrical stimulation of the sural nerve at the ankle was continuously recorded from the ipsilateral biceps femoris. RESULTS: In healthy volunteers, significant progressive inhibition of the RIII reflex was observed during slow ramp distension (61 (13)% of control values) while biphasic effects (facilitation and inhibition) were observed during rapid distensions. In contrast, in IBS patients, the RIII reflex was significantly facilitated during slow ramp distension (139 (15)% of control values) and inhibitions induced by rapid distensions were significantly reduced. Volumes of distension and rectal compliance were similar in both groups. CONCLUSIONS: Our results provide direct evidence that a hyperexcitability of spinal nociceptive processes is present in a large subgroup of IBS patients. PMID- 15361497 TI - Cost of illness of inflammatory bowel disease in the UK: a single centre retrospective study. AB - BACKGROUND AND AIMS: The potentially high costs of care associated with inflammatory bowel disease (IBD) are recognised but we have little knowledge of the scale, profile, or determinants of these costs in the UK. This study aimed to describe costs of illness for a group of IBD patients and determine factors associated with increased healthcare costs. SETTING: A university hospital serving a target population of approximately 330 000. PATIENTS AND METHODS: A six month cohort of IBD patients receiving any form of secondary care was identified, comprising 307 cases of ulcerative (or indeterminate) colitis and 172 cases of Crohn's disease. Demographic and clinical data were abstracted from clinical records and individual resource use was itemised for all attributable costs (including extraintestinal manifestations). Item costs were derived from national and local sources. Cost data were expressed as mean six month costs per patient (with 95% confidence interval (CI)) obtained using non-parametric bootstrapping. Determinants of cost were analysed using generalised linear regression modelling. A postal survey of patients was undertaken to examine indirect costs, out of pocket expenses, and primary care visits. RESULTS: Inpatient services (medical and/or surgical) were required by 67 patients (14%) but accounted for 49% of total secondary care costs. Drug costs accounted for less than a quarter of total costs. Individual patient costs ranged from 73 to 33,254 UK pounds per six months. Mean (95% CI) six month costs per patient were 1256 UK pounds ( 988 pounds, 1721 pounds) for colitis and 1652 UK pounds (1221 pounds, 2239 pounds) for Crohn's disease. Hospitalisation, disease severity grade, and disease extent correlated positively with cost of illness but costs were independent of age or sex. Compared with quiescent cases of IBD, disease relapse was associated with a 2-3-fold increase in costs for non-hospitalised cases and a 20-fold increase in costs for hospitalised cases. Survey data suggested average six month costs were < 30 UK pounds per patient for primary care visits (both diseases) and median loss of earnings were 239 UK pounds for colitis and 299 UK pounds for Crohn's disease. CONCLUSIONS: This study represents the first detailed characterisation of the scale and determinants of costs of illness for IBD in a British hospital. Hospitalisation affected a minority of sufferers but accounted for half of the total direct costs falling on the healthcare system. PMID- 15361498 TI - Influence of dietary factors on the clinical course of ulcerative colitis: a prospective cohort study. AB - BACKGROUND AND AIMS: The causes of relapses of ulcerative colitis (UC) are unknown. Dietary factors have been implicated in the pathogenesis of UC. The aim of this study was to determine which dietary factors are associated with an increased risk of relapse of UC. METHODS: A prospective cohort study was performed with UC patients in remission, recruited from two district general hospitals, who were followed for one year to determine the effect of habitual diet on relapse. Relapse was defined using a validated disease activity index. Nutrient intake was assessed using a food frequency questionnaire and categorised into tertiles. Adjusted odds ratios for relapse were determined using multivariate logistic regression, controlling for non-dietary factors. RESULTS: A total of 191 patients were recruited and 96% completed the study. Fifty two per cent of patients relapsed. Consumption of meat (odds ratio (OR) 3.2 (95% confidence intervals (CI) 1.3-7.8)), particularly red and processed meat (OR 5.19 (95% CI 2.1-12.9)), protein (OR 3.00 (95% CI 1.25-7.19)), and alcohol (OR 2.71 (95% CI 1.1-6.67)) in the top tertile of intake increased the likelihood of relapse compared with the bottom tertile of intake. High sulphur (OR 2.76 (95% CI 1.19-6.4)) or sulphate (OR 2.6 (95% CI 1.08-6.3)) intakes were also associated with relapse and may offer an explanation for the observed increased likelihood of relapse. CONCLUSIONS: Potentially modifiable dietary factors, such as a high meat or alcoholic beverage intake, have been identified that are associated with an increased likelihood of relapse for UC patients. Further studies are needed to determine if it is the sulphur compounds within these foods that mediates the likelihood of relapse and if reducing their intake would reduce relapse frequency. PMID- 15361499 TI - Late complication of blunt abdominal trauma. PMID- 15361500 TI - CDP571, a humanised monoclonal antibody to tumour necrosis factor alpha, for moderate to severe Crohn's disease: a randomised, double blind, placebo controlled trial. AB - BACKGROUND: Targeting tumour necrosis factor alpha (TNF-alpha) has demonstrated efficacy in Crohn's disease. AIM: To evaluate CDP571, a humanised antibody to TNF alpha, for treating active Crohn's disease. PATIENTS: A total of 396 patients with moderate to severe Crohn's disease. METHODS: In a 28 week, randomised, double blind, placebo controlled trial, patients received intravenous CDP571 (10 mg/kg) or placebo every eight weeks to week 24. The primary outcome measure was clinical response (a decrease in the Crohn's disease activity index (CDAI) to > or =100 points or remission (CDAI score < or =150 points)) at week 28. A secondary outcome measure was clinical response (using the same definition) at week 2. RESULTS: Clinical response occurred at week 28 in 80/263 (30.4%) CDP571 patients and 31/132 (23.5%) placebo patients (p = 0.102). Clinical response at week 2 occurred in 90/263 (34.2%) CDP571 patients and 28/132 (21.2%) placebo patients (p = 0.011). Post hoc exploratory subgroup analysis of 159 patients with baseline C reactive protein (CRP) > or =10 mg/l demonstrated significant differences between CDP571 and placebo in clinical response rates at weeks 2 (CDP571, 50/101 (49.5%); placebo, 9/58 (15.5%); p<0.001) and 28 (CDP571, 29/101 (28.7%); placebo, 7/58 (12.1%); p = 0.018). Adverse events occurred at similar frequencies in both treatment groups. CONCLUSIONS: CDP571 is modestly effective for short but not long term treatment of unselected patients with moderate to severe Crohn's disease. The clinical relevance of this short term effect is unclear. Post hoc analysis suggests both short and long term efficacy of CDP571 in patients with elevated baseline CRP (> or =10 mg/l). CDP571 is well tolerated. PMID- 15361501 TI - Large abdominal mass in Crohn's disease. PMID- 15361502 TI - Genotype C hepatitis B virus infection is associated with an increased risk of hepatocellular carcinoma. AB - BACKGROUND: Identification of risk factors for the development of hepatocellular carcinoma (HCC) is important for HCC surveillance in chronic hepatitis B virus (HBV) infection. Our aim was to study the independent risk factors and effect of HBV genotypes on HCC development in a prospective longitudinal cohort of chronic hepatitis B patients. PATIENTS AND METHODS: Chronic hepatitis B patients recruited since 1997 were prospectively followed up for the development of HCC. HCC was diagnosed by a combination of alpha fetoprotein, imaging, and histology. Liver cirrhosis was defined as ultrasonic features of cirrhosis together with hypersplenism, ascites, varices, and/or encephalopathy. RESULTS: In total, 426 patients were followed up for 1664 person years; median 225 (range 12-295) weeks. Forty nine (11%) patients had underlying clinical liver cirrhosis. A total of 242 (57%) and 179 (42%) patients had HBV genotypes C and B, respectively. Twenty five patients developed HCC in a median follow up of 121 (range 14-236) weeks. The overall incidence of HCC was 1502 cases per 100 000 person years. On multivariate analysis, clinical liver cirrhosis and HBV genotype C infection were independently associated with HCC development, with an adjusted relative risk of 10.24 (95% confidence interval (CI) 4.39-23.89; p<0.001) and 2.84 (95% CI 1.05 7.72; p = 0.040), respectively. Patient age, sex, hepatitis B e antigen (HBeAg) status, alanine aminotransferase (ALT) levels, and basal core promoter mutations did not predict HCC development. Patients infected with HBV genotype C tended to have persistently positive HBeAg or fluctuating HBeAg status and higher ALT levels during the follow up period. CONCLUSION: Genotype C HBV infection is an independent risk factor for HCC development in addition to liver cirrhosis. PMID- 15361503 TI - Survey of hepatitis B surface variant infection in children 15 years after a nationwide vaccination programme in Taiwan. AB - BACKGROUND: It is not known whether hepatitis B virus (HBV) with mutations in the a determinant (amino acids (aa) 121-149) of the hepatitis B surface antigen (HBsAg) affect vaccination efficacy. AIM: To investigate the prevalence and clinical significance of these mutants in children, 15 years after universal vaccination in Taiwan. METHODS: Nucleotide sequences encoding the a determinant region (aa 110-160) of HBsAg were analysed in all HBV-DNA positive sera from 1357 children and 219 adolescents serosurveyed in 1999. We then compared the prevalence and changes in the mutants in these children with our previous surveys in the same area conducted in 1984 (just before vaccination), 1989, and 1994. RESULTS: The prevalence of a determinant mutants in HBV-DNA positive children was 7.8% (8/103) in 1984, which significantly increased to 19.6% (10/51) in 1989, peaked at 28.1% (9/32) in 1994, and remained at 23.1% ((3/13) (T131I, G145R, G145R)) in 1999; it was higher in those fully vaccinated compared with those not vaccinated (15/46 v 15/153; p<0.001). However, the number of mutant infected children in each survey was stable in the first 5-10 year period but decreased 10 15 years post vaccination. Increased amino acid variation in the a determinant region occurred in carrier children in the post vaccination survey. Mutated residues tended to occur more frequently in the region with greater local hydrophilicity (residues 140-149) in those vaccinated than in unvaccinated children with variant infection (12/15 v 6/15; p = 0.062). More HBsAg positive a determinant mutants emerged in children fully vaccinated with plasma derived vaccine than those given recombinant vaccine (10/2399 (0.46%) v 0/503; p = 0.122). CONCLUSION: We found that a determinant variants have an advantage in infecting immunised children but do not threaten current HBV vaccination strategies in Taiwan. PMID- 15361504 TI - Long term clinical outcome of chronic hepatitis C patients with sustained virological response to interferon monotherapy. AB - BACKGROUND: The key end point for treatment efficacy in chronic hepatitis C is absence of detectable virus at six months after treatment. However, the incidence of clinical events during long term follow up of patients with sustained virological response is still poorly documented and may differ between the Eastern and Western world. AIMS: To assess clinical end points during long term follow up of European patients with a sustained virological response to interferon monotherapy. METHODS: Meta-analysis of individual patient data from eight European protocolled follow up studies of interferon treatment for chronic hepatitis C. RESULTS: A total of 286 sustained virological responders and 50 biochemical responders (detectable virus but normal alanine aminotransferase levels) were followed up for 59 months. Fifteen sustained virological responders (5.2%) had cirrhosis before treatment and 112 (39%) had genotype 1. The late virological relapse rate after five years of follow up was 4.7% (95% confidence interval (CI) 2.0-7.4) among sustained virological responders; all late relapses occurred within four years after treatment. Among sustained virological responders, the rate of decompensation after five years of follow up was 1.0% (95% CI 0.0-2.3) and none developed hepatocellular carcinoma (HCC). Survival was comparable with the general population, matched for age and sex, the standard mortality ratio being 1.4 (95% CI 0.3-2.5). Clinical outcome of patients with cirrhosis was similar to other sustained virological responders. For biochemical responders, the rates of development of decompensation and HCC during long term follow up were 9.1% (95% CI 0.5-17.7) and 7.1% (95% CI 0-15.0), respectively. CONCLUSIONS: Five year survival of European sustained virological responders was similar to the overall population, matched for age and sex. No HCCs were detected during long term follow up. PMID- 15361505 TI - Delayed hepcidin response explains the lag period in iron absorption following a stimulus to increase erythropoiesis. AB - INTRODUCTION: The delay of several days between an erythropoietic stimulus and the subsequent increase in intestinal iron absorption is commonly believed to represent the time required for body signals to programme the immature crypt enterocytes and for these cells to migrate to the villus. Recent data however suggest that signals from the body to alter absorption are mediated by circulating hepcidin and that this peptide exerts its effect on mature villus enterocytes. METHODS: We have examined the delay in the absorptive response following stimulated erythropoiesis using phenylhydrazine induced haemolysis and correlated this with expression of hepcidin in the liver and iron transporters in the duodenum. RESULTS: There was a delay of four days following haemolysis before a significant increase in iron absorption was observed. Hepatic hepcidin expression did not decrease until day 3, reaching almost undetectable levels by days 4 and 5. This coincided with the increase in duodenal expression of divalent metal transporter 1, duodenal cytochrome b, and Ireg1. CONCLUSION: These results suggest that the delayed increase in iron absorption following stimulated erythropoiesis is attributable to a lag in the hepcidin response rather than crypt programming, and are consistent with a direct effect of the hepcidin pathway on mature villus enterocytes. PMID- 15361506 TI - Association of idiopathic hepatic sinusoidal dilatation with the immunological features of the antiphospholipid syndrome. AB - BACKGROUND: Isolated sinusoidal dilatation is an uncommon hepatic lesion and the cause is largely unknown. OBJECTIVE: To investigate whether prothrombotic disorders or perisinusoidal cell changes could be involved in pure idiopathic hepatic sinusoidal dilatation (HSD). METHODS: Evaluation for associated conditions, prothrombotic disorders, and studies of hepatic perisinusoidal cell activation in consecutive patients, seen between 1993 and 2002, with isolated sinusoidal dilatation unrelated to outflow block, sinusoidal infiltration, or hepatic granulomas. RESULTS: Among 11 patients, associated conditions were prothrombotic disorders (n = 5) and oral contraceptive use (n = 3). Prothrombotic disorders were polycythemia vera (n = 1) and anticardiolipin antibodies combined with lupus anticoagulant (n = 4). No genetic thrombophilia factor was found. Of four patients with lupus anticoagulant, three had antinuclear factors and high serum levels of anticardiolipin antibodies at repeated testing. There was no evidence of intrahepatic or extrahepatic thrombosis in any of the patients. Sinusoidal dilatation was marked in six of 11 patients (54%), including two patients with antiphospholipid antibodies. Activated perisinusoidal cells were only found around markedly dilated sinusoids. CONCLUSION: Idiopathic pure HSD is frequently associated with the immunological features of the antiphospholipid syndrome. Therefore, finding pure HSD in a liver biopsy specimen should prompt the search for antiphospholipid antibodies. PMID- 15361508 TI - Oesophageal motor functions and its disorders. PMID- 15361509 TI - Antibiotic use and the development of Crohn's disease: methodological issues. PMID- 15361507 TI - Pharmacology of serotonin: what a clinician should know. PMID- 15361510 TI - Enhanced vascular responses to noradrenaline in isolated omental arteries from patients with advanced cirrhosis: evidence against intrinsic vascular hyporeactivity in cirrhosis. PMID- 15361511 TI - Histopathology using the Vienna criteria: clinical decision making is still adequate. PMID- 15361512 TI - Population based screening for coeliac disease: patient's choice or doctor's decision. PMID- 15361513 TI - The enteric nervous system is not essential for the propulsion of gut contents in fetal mice. PMID- 15361514 TI - Risk of duodenal cancer in patients with familial adenomatous polyposis. PMID- 15361515 TI - Diaphragm electromyogram root mean square response to hypercapnia and its intersubject and day-to-day variation. AB - Diaphragm activation can be quantified by measuring the root mean square of crural EMG (RMSdi) (Beck J, Sinderby C, Lindstrom L, and Grassino A, J Appl Physiol 85: 1123-1134, 1998). To examine intersubject and day-to-day variation in the RMSdi-Pco(2) relationship, end-tidal Pco(2), minute ventilation (Ve), respiratory frequency (f(B)), and RMSdi were measured in seven healthy subjects on two occasions during steady-state ventilation at seven levels of inspired O(2) fraction (Fi(CO(2))) from 0 to 0.08 in random order. RMSdi was measured with a multielectrode esophageal catheter and controlled for signal contamination and diaphragm position. RMSdi was normalized for values obtained during quiet breathing at functional residual capacity, at Fi(CO(2)) of 0.04, and during an inspiratory capacity maneuver (RMSdi%max) as well as ECG R-wave amplitude at functional residual capacity (RMSdi/ECG(R)), f(B), and thickness of the costal diaphragm measured by ultrasound. RMSdi increased linearly with Pco(2) (mean r(2) = 0.83 +/- 0.10); at the highest Fi(CO(2)), RMSdi%max was 40.2 +/- 11.6%. Relative to the intersubject variation in the Ve-Pco(2) relationship, intersubject variations in the slopes and intercepts of the RMSdi-Pco(2) relationships were 1.7 and 1.8 times, respectively, and RMSdi%max-Pco(2) relationships 0.9 and 1.3 times, respectively, and were unrelated to f(B) and diaphragm thickness. Relative to the day-to-day variation in the Ve-Pco(2) relationship, day-to-day variation in the slopes and intercepts of the RMSdi Pco(2) relationships were 2.8 and 4.4 times, respectively, and RMSdi/ECG(R) Pco(2) relationships 1.3 and 2.2 times, respectively. It was concluded that the RMSdi-Pco(2) relationship measures chemosensitivity and is best compared between subjects via RMSdi%max and on separate occasions in the same subject via RMSdi/ECG(R). PMID- 15361516 TI - Skeletal muscle adaptation: training twice every second day vs. training once daily. AB - Low muscle glycogen content has been demonstrated to enhance transcription of a number of genes involved in training adaptation. These results made us speculate that training at a low muscle glycogen content would enhance training adaptation. We therefore performed a study in which seven healthy untrained men performed knee extensor exercise with one leg trained in a low-glycogen (Low) protocol and the other leg trained at a high-glycogen (High) protocol. Both legs were trained equally regarding workload and training amount. On day 1, both legs (Low and High) were trained for 1 h followed by 2 h of rest at a fasting state, after which one leg (Low) was trained for an additional 1 h. On day 2, only one leg (High) trained for 1 h. Days 1 and 2 were repeated for 10 wk. As an effect of training, the increase in maximal workload was identical for the two legs. However, time until exhaustion at 90% was markedly more increased in the Low leg compared with the High leg. Resting muscle glycogen and the activity of the mitochondrial enzyme 3-hydroxyacyl-CoA dehydrogenase increased with training, but only significantly so in Low, whereas citrate synthase activity increased in both Low and High. There was a more pronounced increase in citrate synthase activity when Low was compared with High. In conclusion, the present study suggests that training twice every second day may be superior to daily training. PMID- 15361517 TI - Cerebral pressure-flow relations in hypertensive elderly humans: transfer gain in different frequency domains. AB - The dynamics of the cerebral vascular response to blood pressure changes in hypertensive humans is poorly understood. Because cerebral blood flow is dependent on adequate perfusion pressure, it is important to understand the effect of hypertension on the transfer of pressure to flow in the cerebrovascular system of elderly people. Therefore, we examined the effect of spontaneous and induced blood pressure changes on beat-to-beat and within-beat cerebral blood flow in three groups of elderly people: normotensive, controlled hypertensive, and uncontrolled hypertensive subjects. Cerebral blood flow velocity (transcranial Doppler), blood pressure (Finapres), heart rate, and end-tidal CO(2) were measured during the transition from a sit to stand position. Transfer function gains relating blood pressure to cerebral blood flow velocity were assessed during steady-state sitting and standing. Cerebral blood flow regulation was preserved in all three groups by using changes in cerebrovascular resistance, transfer function gains, and the autoregulatory index as indexes of cerebral autoregulation. Hypertensive subjects demonstrated better attenuation of cerebral blood flow fluctuations in response to blood pressure changes both within the beat (i.e., lower gain at the cardiac frequency) and in the low-frequency range (autoregulatory, 0.03-0.07 Hz). Despite a better pressure autoregulatory response, hypertensive subjects demonstrated reduced reactivity to CO(2). Thus otherwise healthy hypertensive elderly subjects, whether controlled or uncontrolled with antihypertensive medication, retain the ability to maintain cerebral blood flow in the face of acute changes in perfusion pressure. Pressure regulation of cerebral blood flow is unrelated to cerebrovascular reactivity to CO(2). PMID- 15361518 TI - Effects of transdifferentiation and EGF on claudin isoform expression in alveolar epithelial cells. AB - Rat alveolar epithelial type II cells grown on polycarbonate filters form high resistance monolayers and concurrently acquire many phenotypic properties of type I cells. Treatment with EGF has previously been shown to increase transepithelial resistance across alveolar epithelial cell (AEC) monolayers. We investigated changes in claudin expression in primary cultured AEC during transdifferentiation to the type I cell-like phenotype (days 0, 1, and 8), and on day 5 in culture +/- EGF (10 ng/ml) from day 0 or day 4. Claudins 4 and 7 were increased, whereas claudins 3 and 5 were decreased, on later compared with earlier days in culture. Exposure to EGF led to increases in claudins 4 and 7 and decreases in claudins 3 and 5. Claudin 1 was only faintly detectable in freshly isolated type II cells and remained unchanged over time in culture and after exposure to EGF. These results suggest that increases in transepithelial resistance accompanying AEC transdifferentiation and/or EGF exposure are mediated, at least in part, by changes in the pattern of expression of specific claudin isoforms. PMID- 15361519 TI - Aging does not alter the mechanosensitivity of the p38, p70S6k, and JNK2 signaling pathways in skeletal muscle. AB - The capacity for skeletal muscle to recover its mass following periods of unloading (regrowth) has been reported to decline with age. Although the mechanisms responsible for the impaired regrowth are not known, it has been suggested that aged muscles have a diminished capacity to sense and subsequently respond to a given amount of mechanical stimuli (mechanosensitivity). To test this hypothesis, extensor digitorum longus muscles from young (2-3 mo) and old (26-27 mo) mice were subjected to intermittent 15% passive stretch (ex vivo) as a source of mechanical stimulation and analyzed for alterations in the phosphorylation of stress-activated protein kinase (p38), ribosomal S6 kinase (p70S6k), and the p54 jun N-terminal kinase (JNK2). The results indicated that the average magnitude of specific tension (mechanical stimuli) induced by 15% stretch was similar in muscles from young and old mice. Young and old muscles also revealed similar increases in the magnitude of mechanically induced p38, p70S6k (threonine/serine 421/424 and threonine 389), and JNK2 phosphorylation. In addition, coincubation experiments demonstrated that the release of locally acting growth factors was not sufficient for the induction of JNK2 phosphorylation, suggesting that JNK2 was activated by a mechanical rather than a mechanical/growth factor-dependent mechanism. Taken together, the results of this study demonstrate that aging does not alter the mechanosensitivity of the p38, p70S6k, and JNK2 signaling pathways in skeletal muscle. PMID- 15361520 TI - p51/p63 Controls subunit alpha3 of the major epidermis integrin anchoring the stem cells to the niche. AB - p51/p63, a member of the tumor suppressor p53 gene family, is crucial for skin development. We describe here identification of ITGA3 encoding integrin alpha(3) as a target of its trans-activating function, proposing that p51/p63 allows epidermal stem cells to express laminin receptor alpha(3)beta(1) for anchorage to the basement membrane. When activated by genotoxic stress or overexpressed ectopically in non-adherent cells, p51/p63 transduced a phenotype to attach to extracellular matrices, which was accompanied by expression of ITGA3. Motifs matching the p53-binding consensus sequence were located in a scattered form in intron 1 of human ITGA3, and served as p51/p63-responsive elements in reporter assays. In addition to the trans-activating ability of the TA isoform, we detected a positive effect of the DeltaN isoform on ITGA3. The high level alpha(3) production in human keratinocyte stem cells diminished upon elimination of p51/p63 by small interfering RNA or by Ca(2+)-induced differentiation. Furthermore, a chromatin immunoprecipitation experiment indicated a physical interaction of p51/p63 with intron 1 of ITGA3. This study provides a molecular basis for the standing hypothesis that p51/p63 is essential for epidermal mesenchymal interactions. PMID- 15361521 TI - Semihemoglobins, high oxygen affinity dimeric forms of human hemoglobin respond efficiently to allosteric effectors without forming tetramers. AB - Significant reduction in oxygen affinity resulting from interactions between heterotropic allosteric effectors and hemoglobin in not only the unligated derivative but also the fully ligated form has been reported (Tsuneshige, A., Park, S. I., and Yonetani, T. (2002) Biophys. Chem. 98, 49-63; Yonetani, T., Park, S. I., Tsuneshige, A., Imai, K., and Kanaori, K. (2002) J. Biol. Chem. 277, 34508-34520). To further investigate this effect in more detail, alpha- and beta semihemoglobins, namely, alpha(heme)beta(apo) and alpha(apo)beta(heme), respectively, were prepared and characterized with respect to the impact of allosteric effectors on both conformation and ligand binding properties. Semihemoglobins are dimers characterized by a high affinity for oxygen and lack of cooperativity. We found that, compared with stripped conditions, semihemoglobins responded to effectors (inositol hexaphosphate and L35) by decreasing the affinity for oxygen by 60- and 130-fold for alpha- and beta semihemoglobins, respectively. 1H NMR and sedimentation velocity experiments carried out with their ligated and unligated forms in the absence and presence of effectors revealed that semihemoglobins always remain as single-heme-carrying dimers. Recombination kinetics of their photolyzed CO derivatives showed that effectors did indeed interact with their ligated forms. Measurements of the Fe His stretching mode show that the semihemoglobins undergo a large ligand binding induced conformational shift and that both ligand-free and ligand derivatives respond to the presence of effectors. Contradictions to the Monod-Wyman Changeaux/Perutz allosteric model arise since 1) the modulation of ligand affinity is not achieved in semihemoglobins by the formation of a low affinity T conformation (quaternary effect) but by direct interaction with effectors, 2) effectors do interact significantly with ligated forms of high affinity semihemoglobins, and 3) modulation of the ligand affinity and the cooperativity are not necessarily linked but instead can be separated into two distinct phenomena that can be isolated. PMID- 15361522 TI - Are respiratory enzymes the primary sources of intracellular hydrogen peroxide? AB - Endogenous H2O2 is believed to be a source of chronic damage in aerobic organisms. To quantify H2O2 formation, we have generated strains of Escherichia coli that lack intracellular scavenging enzymes. The H2O2 that is formed within these mutants diffuses out into the medium, where it can be measured. We sought to test the prevailing hypothesis that this H2O2 is primarily generated by the autoxidation of redox enzymes within the respiratory chain. The rate of H2O2 production increased when oxygen levels were raised, confirming that H2O2 is formed by an adventitious chemical process. However, mutants that lacked NADH dehydrogenase II and fumarate reductase, the most oxidizable components of the respiratory chain in vitro, continued to form H2O2 at normal rates. NADH dehydrogenase II did generate substantial H2O2 when it was when overproduced or quinones were absent, forcing electrons to accumulate on the enzyme. Mutants that lacked both NADH dehydrogenases respired very slowly, as expected; however, these mutants showed no diminution of H2O2 excretion, suggesting that H2O2 is primarily formed by a source outside the respiratory chain. That source has not yet been identified. In respiring cells the rate of H2O2 production was approximately 0.5% the rate of total oxygen consumption, with only modest changes when cells used different carbon sources. PMID- 15361523 TI - Ex vivo analysis of lactate and glucose metabolism in the rat brain under different states of depressed activity. AB - Brain metabolism of glucose and lactate was analyzed by ex vivo NMR spectroscopy in rats presenting different cerebral activities induced after the administration of pentobarbital, alpha-chloralose, or morphine. The animals were infused with a solution of either [1-(13)C]glucose plus lactate or glucose plus [3-(13)C]lactate for 20 min. Brain metabolite contents and enrichments were determined from analyses of brain tissue perchloric acid extracts according to their post-mortem evolution kinetics. When amino acid enrichments were compared, both the brain metabolic activity and the contribution of blood glucose relative to that of blood lactate to brain metabolism were linked with cerebral activity. The data also indicated the production in the brain of lactate from glycolysis in a compartment other than the neurons, presumably the astrocytes, and its subsequent oxidative metabolism in neurons. Therefore, a brain electrical activity-dependent increase in the relative contribution of blood glucose to brain metabolism occurred via the increase in the metabolism of lactate generated from brain glycolysis at the expense of that of blood lactate. This result strengthens the hypothesis that brain lactate is involved in the coupling between neuronal activation and metabolism. PMID- 15361524 TI - Rodent and human mast cells produce functionally significant intracellular reactive oxygen species but not nitric oxide. AB - In immunity, reactive oxygen species (ROS) and nitric oxide (NO) are important antimicrobial agents and regulators of cell signaling and activation pathways. However, the cellular sources of ROS and NO are much debated. Particularly, there is contention over whether mast cells, key secretory cells in allergy and immunity, can generate these chemical species, and if so, whether they are of functional significance. We therefore examined directly by flow cytometry the capacity of mast cells to generate intracellular ROS and NO using the respective cell-permeable fluorescent probes dichlorodihydrofluorescein and diaminofluorescein and evaluated the effects of inhibitors of ROS and NO synthesis on cell degranulation. For each of three mast cell types (rat peritoneal mast cells, mouse bone marrow-derived mast cells, and human blood derived mast cells), degranulation stimulated by IgE/antigen was accompanied by production of intracellular ROS but not NO. Inhibition of ROS production led to reduced degranulation, indicating a facilitatory role for ROS, whereas NO synthase inhibitors were without effect. Likewise, bacterial lipopolysaccharide and interferon-gamma over a wide range of conditions failed to generate intracellular NO in mast cells, whereas these agents readily induced intracellular NO in macrophages. NO synthase protein, as assessed by Western blotting, was readily induced in macrophages but not mast cells. We conclude that rodent and human mast cells generate intracellular ROS but not NO and that intracellular ROS but not intracellular NO are functionally linked to mast cell degranulation. PMID- 15361525 TI - Positive contribution of hydration on DNA binding by E2c protein from papillomavirus. AB - Protein-nucleic acid interactions are responsible for the regulation of key biological events such as genomic transcription and recombination and viral replication. However, the recognition mechanisms involved in these processes are not completely understood. Here, we investigate the dominant forces involved in protein-protein and protein-DNA interactions for the 80-amino-acid C-terminal domain of the E2 protein (E2c) from human papillomavirus (HPV-16). The E2c protein is a homodimer that specifically binds to double-stranded DNA containing the consensus sequence ACCG-N(4)-CGGT, where N is any nucleotide. DNA binding affinity is reduced by lowering water chemical potential, accompanied by an increase in cooperativity. Wyman linkage relations between affinity and water chemical potential indicate that 11 additional water molecules are bound in the formation of the complex between E2c and DNA. Salt dissociation isotherms showed that 10 counterions are released upon association, even at low water activity, indicating that this latter variable does not change the electrostatic component of the interaction. Further analysis demonstrates a strong dependence of cooperativity of binding on the protein concentration. Altogether, these results reveal a novel binding pathway in which the consolidated complex may achieve its final form via a monomer-DNA intermediate, which favors the binding of a second monomer. This molecular mechanism reveals the contributions of multiple conformers in a tight virus genome modulation that seems to be important in the cell infection scenario. PMID- 15361526 TI - Controlled alternate partial root-zone irrigation: its physiological consequences and impact on water use efficiency. AB - Controlled alternate partial root-zone irrigation (CAPRI), also called partial root-zone drying (PRD) in other literature, is a new irrigation technique and may improve the water use efficiency of crop production without significant yield reduction. It involves part of the root system being exposed to drying soil while the remaining part is irrigated normally. The wetted and dried sides of the root system are alternated with a frequency according to soil drying rate and crop water requirement. The irrigation system is developed on the basis of two theoretical backgrounds. (i) Fully irrigated plants usually have widely opened stomata. A small narrowing of the stomatal opening may reduce water loss substantially with little effect on photosynthesis. (ii) Part of the root system in drying soil can respond to the drying by sending a root-sourced signal to the shoots where stomata may be inhibited so that water loss is reduced. In the field, however, the prediction that reduced stomatal opening may reduce water consumption may not materialize because stomatal control only constitutes part of the total transpirational resistance. The boundary resistance from the leaf surface to the outside of the canopy may be so substantial that reduction in stomatal conductance is small and may be partially compensated by the increase in leaf temperature. It is likely that densely populated field crops, such as wheat and maize, may have a different stomatal control over transpiration from that of fruit trees which are more sparsely separated. It was discussed how long the stomata can keep 'partially' closed when a prolonged and repeated 'partial' soil drying is applied and what role the rewatering-stimulated new root growth may play in sensing the repeated soil drying. The physiological and morphological alternation of plants under partial root-zone irrigation may bring more benefits to crops than improved water use efficiency where carbon redistribution among organs is crucial to the determination of the quantity and quality of the products. PMID- 15361527 TI - Agronomic options for improving rainfall-use efficiency of crops in dryland farming systems. AB - Yields of dryland (rainfed) wheat in Australia have increased steadily over the past century despite rainfall being unchanged, indicating that the rainfall-use efficiency has increased. Analyses suggest that at least half of the increase in rainfall-use efficiency can be attributed to improved agronomic management. Various methods of analysing the factors influencing dryland yields and rainfall use efficiency, such as simple rules and more complex models, are presented and the agronomic factors influencing water use, water-use efficiency, and harvest index of crops are discussed. The adoption of agronomic procedures such as minimum tillage, appropriate fertilizer use, improved weed/disease/insect control, timely planting, and a range of rotation options, in conjunction with new cultivars, has the potential to increase the yields and rainfall-use efficiency of dryland crops. It is concluded that most of the agronomic options for improving rainfall-use efficiency in rainfed agricultural systems decrease water losses by soil evaporation, runoff, throughflow, deep drainage, and competing weeds, thereby making more water available for increased water use by the crop. PMID- 15361528 TI - Perception of Bradyrhizobium japonicum Nod factor by soybean [Glycine max (L.) Merr.] root hairs under abiotic stress conditions. AB - Suboptimal growth conditions, such as low rhizosphere temperature, high salinity, and low pH can negatively affect the rhizobia-legume symbioses, resulting in poor nodulation and lower amounts of nitrogen fixed. Early stages of the Bradyrhizobium japonicum-soybean [Glycine max (L.) Merr.] symbiosis, such as excretion of genistein (the plant-to-bacteria signal) and infection initiation can be inhibited by abiotic stresses; however, the effect on early events modulated by Nod factors (bacteria-to-plant signalling), particularly root hair deformations is unknown. Thus, the objective of this study was to evaluate the perception of Nod factor by soybean root hairs under three stress conditions: low temperature, low pH, and high salinity. Three experiments were conducted using a 1:1 ratio of Nod Bj-V (C(18:1), MeFuc) and Nod Bj-V (Ac, C(16:0), MeFuc). Nod factor induced four types of root hair deformation (HAD), wiggling, bulging, curling, and branching. Under optimal experimental conditions root hair response to the three levels of Nod factor tested (10(-6), 10(-8), and 10(-10) M) was dose dependent. The highest frequency of root hair deformations was elicited by the 10(-6) M level. Root hair deformation decreased with temperature (25, 17, and 15 degrees C), low pH, and high salinity. Nod factor concentration did not interact with either low temperature or pH. However, salinity strongly inhibited HAD responses to increases in Nod factor concentration. Thus, the addition of higher levels of Nod factor is able to overcome the effects of low pH and temperature stress, but not salinity. PMID- 15361529 TI - Senescence and programmed cell death: substance or semantics? AB - The terms senescence and programmed cell death (PCD) have led to some confusion. Senescence as visibly observed in, for example, leaf yellowing and petal wilting, has often been taken to be synonymous with the programmed death of the constituent cells. PCD also obviously refers to cells, which show a programme leading to their death. Some scientists noted that leaf yellowing, if it has not gone too far, can be reversed. They suggested calling leaf yellowing, before the point of no return, 'senescence' and the process after it 'PCD'. However, this runs into several problems. It is counter to the historical definitions of senescence, both in animal and plant science, which stipulate that senescence is programmed and directly ends in death. It would also mean that only leaves and shoots show senescence, whereas several other plant parts, where reversal has not (yet) been shown, have no senescence, but only PCD. This conflicts with ordinary usage (as in root and flower senescence). Moreover, a programme can be reversible and therefore it is not counter to logic to regard the cell death programme as potentially reversible. In green leaf cells a decision to die, in a programmed way, has been taken, in principle, before the cells start to remobilize their contents (that is, before visible yellowing) and only rarely is this decision reversed. According to the arguments developed here there are no good reasons to separate a senescence phase and a subsequent PCD phase. Rather, it is asserted, senescence in cells is the same as PCD and the two are fully synchronous. PMID- 15361530 TI - Nod factor-treated Medicago truncatula roots and seeds show an increased number of nodules when inoculated with a limiting population of Sinorhizobium meliloti. AB - Medicago truncatula is a model legume plant that interacts symbiotically with Sinorhizobium meliloti, the alfalfa symbiont. This process involves a molecular dialogue between the bacterium and the plant. Legume roots exude flavonoids that induce the expression of a set of rhizobial genes, the nod genes, which are essential for nodulation and determination of the host range. In turn, nod genes control the synthesis of lipo-chito-oligosaccharides (LCOs), Nod factors, which are bacteria-to-plant signal molecules mediating recognition and nodule organogenesis. M. truncatula roots or seeds have been treated with Nod factors and hydroponically growing seedlings have been inoculated with a limiting population of S. meliloti. It has been shown that submicromolar concentrations of Nod factors increase the number of nodules per plant on M. truncatula. Compared with roots, this increase is more noticeable when seeds are treated. M. truncatula seeds are receptive to submicromolar concentrations of Nod factors, suggesting the possibility of a high affinity LCO perception system in seeds or embryos as well. PMID- 15361531 TI - Alterations in pyrimidine nucleotide metabolism as an early signal during the execution of programmed cell death in tobacco BY-2 cells. AB - Changes in pyrimidine metabolism were investigated during programmed cell death (PCD) of tobacco BY-2 cells, induced by a simultaneous increase in the endogenous levels of nitric oxide (NO) and hydrogen peroxide. The de novo synthesis of pyrimidine nucleotides was estimated by following the metabolic fate of the (14)C labelled orotic acid, whereas the rates of salvage and degradation pathways were studied by measuring the respective incorporation of (14)C-labelled uridine and uracil under different treatments. Nucleic acid metabolism was also examined using labelled thymidine as a marker. The results show that specific alterations in the balance of pyrimidine nucleotide synthesis, which include a decreased rate of salvage activity of uracil and uridine and increased salvage activity of thymidine, represent a metabolic switch that establishes proper cellular conditions for the induction of PCD. In particular, a reduction in the utilization of uracil for salvage products occurs very early during PCD, before the appearance of typical cytological features of the death programme, thus representing an early metabolic marker for PCD. These changes are strictly associated with PCD, since they do not occur if NO or hydrogen peroxide are increased individually, or if actinomycin, which inhibits the death programme, is added into the medium in the presence of NO and hydrogen peroxide. The possible roles of these fluctuations in pyrimidine metabolism on the cellular nucleotide pool are discussed in relation to the induction of cell death. PMID- 15361532 TI - Ion flux interaction with cytoplasmic streaming in branchlets of Chara australis. AB - Both parts of the actin-myosin complex involved in cytoplasmic streaming could be regulated by mineral ions. The main goal of this study was to find a relationship between cyclosis and ion transport across the cell wall and plasma membrane. The transport of K(+) and Ca(2+) along pH bands in Chara branchlet internodal cells was characterized by using the MIFE system for non-invasive microelectrode measurement of ion fluxes. Branchlets formed acidic and alkaline bands with the pH ranging from 5 to 8. Different pH patterns were observed for different sides of the branchlets. Sides with cyclosis streaming acropetally generally showed greater variation in the profiles of pH and H(+) fluxes. Although a high correlation was not found between pH bands and Ca(2+) or K(+) fluxes, there was a positive correlation between Ca(2+) and K(+) fluxes themselves for both sides of the branchlets. Application of cytochalasin D, an inhibitor of cyclosis, had no immediate effect on pH and ion fluxes, however, the time of cyclosis cessation corresponded with a dramatic change in Ca(2+) and K(+) fluxes; pH profiles and H(+) fluxes were affected within 2 h. The evidence suggests that, in Chara branchlets, pH band formation and Gd(3+)-insensitive Ca(2+) transport systems are linked to the cyclosis machinery: (i) the pH band amplitude for the acropetally streaming side was larger than that for the basipetally streaming side; (ii) cessation of cytoplasmic streaming after cytochalasin D application resulted in changed pH banding profiles and H(+), Ca(2+) and K(+) fluxes; and (iii) the application of GdCl(3) or incubation in GdCl(3) solutions did not lead to the cessation of cytoplasmic streaming, although external Ca(2+) fluxes changed. PMID- 15361533 TI - Differential accumulation of Cd in durum wheat cultivars: uptake and retranslocation as sources of variation. AB - Durum wheat (Triticum turgidum L. var. durum) accumulates Cd from the soil depending on various factors. When grown in hydroponic solution containing Cd (20 microg l(-1)), roots had higher tissue Cd concentrations than shoots or heads. Kyle (the higher grain-Cd accumulating cultivar) had lower root-Cd, and greater shoot-Cd and head-Cd concentrations than Arcola (the lower grain-Cd accumulating cultivar). These cultivar differences were greater at flowering and ripening than at tillering. Much of the root-Cd was lost between the flowering and ripening stages of development. Distribution of (106)Cd among plant parts, after a single 24 h feeding, demonstrated that root-to-shoot transfer of Cd in Arcola was similar to that of Kyle at tillering, but it had ceased at flowering in Arcola but not Kyle. None of the Cd in wheat heads at ripening originated from (106)Cd exposure in the previous 24 h, suggesting that grain-Cd is a function of total shoot accumulation. Both cultivars demonstrated basipetal translocation of Cd; Arcola at tillering translocated more Cd from shoots to roots than Kyle. The proportion of Cd(2+)/Cd(total) in the nutrient solution decreased with time, suggesting that plant activity altered the solution chemistry. The alteration probably resulted from either preferential depletion of solution Cd(2+) and/or addition of root exudates. Lower grain-Cd accumulation in Arcola possibly resulted from a combination of reduced root-to-shoot transfer of Cd at flowering, as well as enhanced shoot-to-root retranslocation of Cd, at least in younger plants. Plant-mediated changes in solution-Cd speciation did not play a role. PMID- 15361534 TI - SOR1, a gene associated with bioherbicide production in sorghum root hairs. AB - Sorghum [Sorghum bicolor (L.) Moench] roots exude a potent bioherbicide known as sorgoleone, which is produced in living root hairs and is phytotoxic to broadleaf and grass weeds at concentrations as low as 10 microM. Differential gene expression was studied in sorghum (S. bicolorxS. sudanense) cv. SX17 between roots with abundant root hairs and those without root hairs using a modified differential display approach. A differentially expressed gene, named SOR1, was cloned by using Rapid Amplification of the 5' ends of cDNA (5'-RACE). Real-time PCR analysis of multiple tissues of sorghum SX17 revealed that the SOR1 transcript level in root hairs was more than 1000 times higher than that of other tissues evaluated, including immature leaf, mature leaf, mature stem, panicle, and roots with hairs removed. Semi-quantitative RT-PCR revealed that SOR1 was expressed in the sorgoleone-producing roots of sorghum SX17, shattercane [S. bicolor (L.) Moench], and johnsongrass [S. halepense (L.) Pers.], but not in the shoots of sorghum or in the roots of sweet corn (Zea mays L.) 'Summer Flavor 64Y', in which sorgoleone production was not detected by HPLC analysis. Similarity searches indicated that SOR1 probably encodes a novel desaturase, which might be involved in the formation of a unique and specific double bonding pattern within the long hydrocarbon tail of sorgoleone. PMID- 15361535 TI - Evidence that the hexose-to-sucrose ratio does not control the switch to storage product accumulation in oilseeds: analysis of tobacco seed development and effects of overexpressing apoplastic invertase. AB - Wild-type tobacco (Nicotiana tabacum L.) seed development was characterized with respect to architecture and carbohydrate metabolism. Tobacco seeds accumulate oil and protein in the embryo, cellular endosperm and inner layer of the seed coat. They have high cell wall invertase (INV) and hexoses in early development which is typical of seeds. INV and the ratio of hexose to sucrose decline during development, switching from high hex to high suc, but not until most oil and all protein accumulation has occurred. The oil synthesis which coincides with the switch is mostly within the embryo. INV activity is greater than sucrose synthase activity throughout development, and both activities exceed the demand for carbohydrate for dry matter accumulation. To investigate the role of INV-mediated suc metabolism in oilseeds, genes for yeast INV and/or hexokinase (HK) were expressed under a seed-specific napin promoter, targeting activity to the apoplast and cytosol, respectively. Manipulating the INV pathway in an oilseed could either increase oil accumulation and sink strength, or disrupt carbohydrate metabolism, possibly through sugar-sensing, and decrease the storage function. Neither effect was found: transgenics with INV and/or HK increased 30-fold and 10 fold above wild-type levels had normal seed size and composition. This contrasted with dramatic effects on sugar contents in the INV lines. PMID- 15361536 TI - Recent developments in understanding the regulation of starch metabolism in higher plants. AB - This article reviews current knowledge of starch metabolism in higher plants, and focuses on the control and regulation of the biosynthetic and degradative pathways. The major elements comprising the synthetic and degradative pathways in plastids are discussed, and show that, despite present knowledge of the core reactions within each pathway, understanding of how these individual reactions are co-ordinated within different plastid types and under different environmental conditions, is far from complete. In particular, recently discovered aspects of the fine control of starch metabolism are discussed, which indicate that a number of key reactions are controlled by post-translational modifications of enzymes, including redox modulation and protein phosphorylation. In some cases, enzymes of the pathway may form protein complexes with specific functional significance. It is suggested that some of the newly discovered aspects of fine control of the biosynthetic pathway may well apply to many other proteins which are directly and indirectly involved in polymer synthesis and degradation. PMID- 15361537 TI - Osmotic stress in barley regulates expression of a different set of genes than salt stress does. AB - Under high salt conditions, plant growth is severely inhibited due to both osmotic and ionic stresses. In an effort to dissect genes and pathways that respond to changes in osmotic potential under salt stress, the expression patterns were compared of 460 non-redundant salt-responsive genes in barley during the initial phase under osmotic versus salt stress using cDNA microarrays with northern blot and real-time RT-PCR analyses. Out of 52 genes that were differentially expressed under osmotic stress, 11, such as the up-regulated genes for pyrroline-5-carboxylate synthetase, betaine aldehyde dehydrogenase 2, plasma membrane protein 3, and the down-regulated genes for water channel 2, heat shock protein 70, and phospholipase C, were regulated in a virtually identical manner under salt stress. These genes were involved in a wide range of metabolic and signalling pathways suggesting that, during the initial phase under salt stress, several of the cellular responses are mediated by changes in osmotic potential. PMID- 15361538 TI - Respiratory properties and malate metabolism in Percoll-purified mitochondria isolated from pineapple, Ananas comosus (L.) Merr. cv. smooth cayenne. AB - An investigation was made of the respiratory properties and the role of the mitochondria isolated from one phosphoenolpyruvate carboxykinase (PCK)-CAM plant Ananas comosus (pineapple) in malate metabolism during CAM phase III. Pineapple mitochondria showed very high malate dehydrogenase (MDH), and low malic enzyme (ME) and glutamate-oxaloacetate transaminase (GOT) activities. The mitochondria readily oxidized succinate and NADH with high rates and coupling, while they only oxidized NADPH in the presence of Ca(2+). Pineapple mitochondria oxidized malate with low rates under most assay conditions, despite increasing malate concentrations, optimizing pH, providing cofactors such as coenzyme A, thiamine pyrophosphate, and NAD(+), and supplying individually external glutamate or GOT. However, providing glutamate and GOT simultaneously strongly increased the rates of malate oxidation. The OAA easily permeated the mitochondrial membranes to import into or export out of pineapple mitochondria during malate oxidation, but the mitochondria did not consume external Asp or alpha-KG. These results suggest that OAA played a significant role in the mitochondrial malate metabolism of pineapple, in which malate was mainly oxidized by active mMDH to produce OAA which could be exported outside the mitochondria via a malate-OAA shuttle. Cytosolic GOT then consumed OAA by transamination in the presence of glutamate, leading to a large increase in respiration rates. The malate-OAA shuttle might operate as a supporting system for decarboxylation in phase III of PCK-CAM pineapple. This shuttle system may be important in pineapple to provide a source of energy and substrate OAA for cytosolic PCK activity during the day when cytosolic OAA and ATP was limited for the overall decarboxylation process. PMID- 15361539 TI - Methyl-beta-D-glucopyranoside in higher plants: accumulation and intracellular localization in Geum montanum L. leaves and in model systems studied by 13C nuclear magnetic resonance. AB - Using (13)C-NMR, methyl-beta-D-glucopyranoside (MeG) was characterized as a major compound in the leaves of the alpine herb Geum montanum L. MeG continuously accumulated during the life span of G. montanum leaves, and accounted for up to 20% of the soluble carbohydrates in aged overwintering leaves, without being reallocated during senescence. Incubating intact plant tissues, culture cells, and purified organelles with (13)C-labelled substrates showed that MeG was synthesized in the cytosol of cells, directly from glucose and methanol molecules. There was no contribution of the C-1 pathway. MeG was subsequently stored in the vacuole without being re-exported to the cytoplasm. All the dicots tested contained the enzymatic machinery permitting MeG synthesis from methanol and glucose, but the plants accumulating this compound at concentrations higher than 1 micromol g(-1) wet wt were mainly members of the Rosaceae family belonging to the Rosoideae subfamily. It is suggested that the synthesis of MeG may contribute to reduce the accumulation in the cytoplasm of methanol and its derived compounds. PMID- 15361540 TI - Differential expression of putative floral genes in Pharbitis nil shoot apices cultured on glucose compared with sucrose. AB - If, following an inductive treatment of 2 d of continuous darkness, shoot apices of Pharbitis nil are cultured 1 d later on White's medium supplemented with 2% sucrose, they cannot form carpels, but they can if they are cultured on 2% glucose. It was hypothesized that the differential effect of these sugars was because of differential expression of carpel-specific genes. Partial cDNA homologues to the Arabidopsis genes, LEAFY (PnLFY), AGAMOUS (PnAG1/2), and CRABS CLAWS (PnCRC1/2) were cloned. PnLFY was expressed in the shoot apex 1 d following the start of induction and remained higher than in non-induced apices for a further 6 d before exhibiting a major peak of expression on day 7. Peaks of expression of PnAG1 and PnAG2 spanned days 7-11, coinciding with the appearance of stamens and then carpels. The Pharbitis 'PnCRC2' showed greatest homology to Arabidopsis YABBY2 (PnYABBY). Its expression peaked on day 8 when the carpels first appeared. 'PnCRC1' showed greatest homology to Arabidopsis FILAMENTOUS (PnFIL). Its expression was approximately the same in inductive and non-inductive treatments. Apart from PnFIL these partial cDNAs could be used as markers to test the hypothesis concerning differential effects of sucrose and glucose. Cultured shoot apices from induced plants were sampled at weekly intervals. All four genes were expressed more strongly in the glucose compared with the sucrose treatment, most notably at day 17. A more intensive sampling (days 15-19) indicated that PnLFY and PnYABBY exhibited much higher expression on glucose compared with sucrose, most notably on days 15-16 and days 18-19. PMID- 15361541 TI - Root phloem-specific expression of the plasma membrane amino acid proton co transporter AAP3. AB - Amino acids are regarded as the nitrogen 'currency' of plants. Amino acids can be taken up from the soil directly or synthesized from inorganic nitrogen, and then circulated in the plant via phloem and xylem. AtAAP3, a member of the Amino Acid Permease (AAP) family, is mainly expressed in root tissue, suggesting a potential role in the uptake and distribution of amino acids. To determine the spatial expression pattern of AAP3, promoter-reporter gene fusions were introduced into Arabidopsis. Histochemical analysis of AAP3 promoter-GUS expressing plants revealed that AAP3 is preferentially expressed in root phloem. Expression was also detected in stamens, in cotyledons, and in major veins of some mature leaves. GFP-AAP3 fusions and epitope-tagged AAP3 were used to confirm the tissue specificity and to determine the subcellular localization of AtAAP3. When overexpressed in yeast or plant protoplasts, the functional GFP-AAP3 fusion was localized in subcellular organelle-like structures, nuclear membrane, and plasma membrane. Epitope-tagged AAP3 confirmed its localization to the plasma membrane and nuclear membrane of the phloem, consistent with the promoter-GUS study. In addition, epitope-tagged AAP3 protein was localized in endodermal cells in root tips. The intracellular localization suggests trafficking or cycling of the transporter, similar to many metabolite transporters in yeast or mammals, for example, yeast amino acid permease GAP1. Despite the specific expression pattern, knock-out mutants did not show altered phenotypes under various conditions including N-starvation. Microarray analyses revealed that the expression profile of genes involved in amino acid metabolism did not change drastically, indicating potential compensation by other amino acid transporters. PMID- 15361542 TI - Activation of cytotoxic lymphocytes by interferon-alpha: role of oxygen radical producing mononuclear phagocytes. AB - A significant part of the therapeutic benefit of interferon-alpha (IFN-alpha) therapy in malignant diseases and in chronic viral infections is assumed to result from activation of lymphocytes with natural killer (NK) and T cell phenotype. In tumor tissue and in chronically infected tissue, the function and viability of these lymphocytes are frequently impaired. Mononuclear phagocyte (MP)-derived reactive oxygen species (ROS) have been proposed to contribute to the lymphocyte suppression in these tissues. Here, we report that three types of human cytotoxic lymphocytes of relevance to immunoactivation by IFN-alpha, CD3epsilon+/8+/56- T cells, CD3epsilon-/56+ NK cells, and CD3epsilon+/56+ NK/T cells became anergic to IFN-alpha induction of the cell-surface activation marker CD69 after exposure to autologous MPs in vitro. In addition to their incapacity to express CD69, cytotoxic lymphocytes acquired features characteristic of apoptosis after incubation with MPs. The lymphocyte apoptosis and nonresponsiveness to IFN-alpha were prevented by two inhibitors of reduced nicotinamide adenine dinucleotide phosphate oxidase-dependent formation of ROS in MPs, histamine dihydrochloride and diphenylene ionodonium, as well as by catalase, a scavenger of ROS. We conclude that MP-derived ROS may negatively affect IFN-alpha-induced immunostimulation and propose that ROS inhibitors or scavengers may be useful to improve lymphocyte activation during treatment with IFN-alpha. PMID- 15361543 TI - Using molecular tools to dissect the role of Galphas in sensitization of AC1. AB - Short-term activation of Galpha(i/o)-coupled receptors inhibits adenylyl cyclase, whereas persistent activation of Galpha(i/o)-coupled receptors results in a compensatory sensitization of adenylyl cyclase activity after subsequent activation by Galpha(s) or forskolin. Several indirect observations have suggested the involvement of increased Galpha(s)-adenylyl cyclase interactions in the expression of sensitization; however, evidence supporting a direct role for Galpha(s) has not been well established. In the present report, we used two genetic approaches to further examine the role of Galpha(s) in heterologous sensitization of Ca(2+)-sensitive type 1 adenylyl cyclase (AC1). In the first approach, we constructed Galpha(s)-insensitive mutants of AC1 (F293L and Y973S) that retained sensitivity to Ca2+ and forskolin activation. Persistent (2 h) activation of the D2 dopamine receptor resulted in a significant augmentation of basal or Ca(2+)- and forskolin-stimulated AC1 activity; however, sensitization of Galpha(s)-insensitive mutants of AC1 was markedly reduced compared with wild-type AC1. In the second strategy, we examined the requirement of an intact receptor Galpha(s) signaling pathway for the expression of sensitization using dominant negative Galpha(s) mutants (alpha3beta5 G226A/A366S or alpha3beta5 G226A/E268A/A366S) to disrupt D1 dopamine receptor activation of recombinant AC1. D1 dopamine receptor-Galpha(s) signaling was attenuated in the presence of alpha3beta5 G226A/A366S or alpha3beta5 G226A/E268A/A366S, but D2 agonist-induced sensitization of Ca(2+)-stimulated AC1 activity was not altered. Together, the present findings directly support the hypothesis that the expression of sensitization of AC1 involves Galpha(s)-adenylyl cyclase interactions. PMID- 15361544 TI - Alterations in receptor expression or agonist concentration change the pathways gastrin-releasing peptide receptor uses to regulate extracellular signal regulated kinase. AB - G protein-coupled receptors activate extracellular signal-regulated kinases (ERKs) via different pathways in different cell types. In this study, we demonstrate that gastrin-releasing peptide receptor (GRPr) regulates ERK through multiple pathways in a single cell type depending upon receptor expression and agonist concentration. We examined stably transfected BALB/c 3T3 fibroblasts expressing GRPr constructs at different levels and treated the cells with several concentrations of bombesin (BN, a GRPr agonist) to activate a variable number of GRPr per cell. GRPr induced two waves of ERK activation and one wave of ERK inhibition. One wave of activation required an intact GRPr carboxyl-terminal domain (CTD). It peaked 6 min after addition of high BN concentration ([BN]) in cells with high GRPr expression. Another wave of activation was CTD-independent. It peaked 2 to 4 min after BN addition in cells when [BN] and/or GRPr expression were lower. The early wave of ERK activation was more sensitive than the later one to pretreatment with Bisindolylmaleimide I (GF 109203X) (a protein kinase C inhibitor) or hypertonic sucrose. Because these two waves of activation differ in time course, dose-response curve, requirement for GRPr CTD, and sensitivity to inhibitors, they result from different signaling pathways. A third pathway in these cells inhibited ERK phosphorylation 2 min after addition of high [BN] in cells with high GRPr expression. Furthermore, a GRPr-expressing human duodenal cancer cell line showed differential sensitivity to GF 109203X throughout BN induced ERK activation, indicating that GRPr may activate ERK via multiple pathways in cells expressing endogenous GRPr. PMID- 15361545 TI - Preferential Interaction between the dopamine D2 receptor and Arrestin2 in neostriatal neurons. AB - Dopamine D2 receptor interactions with arrestins and arrestin-dependent internalization have been characterized using heterologously expressed D2 receptor and arrestins. The purpose of this study was to investigate D2 receptor interaction with endogenous arrestins. Arrestin2 and arrestin3 in striatal homogenates bound to the third cytoplasmic loop of the D2 receptor, and purified arrestin2 and arrestin3 bound to the second and third loops and C terminus of the D2 receptor, in a glutathione S-transferase pull-down assay. In NS20Y neuroblastoma cells expressing an enhanced green-fluorescent protein-tagged D2 receptor (D2-EGFP), 2-h D2 agonist stimulation enhanced the colocalization of D2 EGFP with endogenous arrestin2 and arrestin3. These results suggest that the D2 receptor has the intrinsic ability to bind both nonvisual arrestins. Agonist treatment of D2-EGFP NS20Y cells induced D2 receptor internalization (36-46%) that was maximal within 20 min, but that was prevented by small interfering RNA induced depletion of arrestin2 and arrestin3. In neostriatal neurons, 2-h agonist treatment selectively increased the colocalization of the endogenous D2 receptor with arrestin2, whereas receptor colocalization with arrestin3 was reduced. Agonist stimulation caused translocation of arrestin2, but not arrestin3, to the membrane in neurons and selectively enhanced the coimmunoprecipitation of the D2 receptor and arrestin2. All three measures of receptor/arrestin interaction (colocalization, translocation, and coprecipitation) demonstrated selective agonist-induced interaction between the D2 receptor and arrestin2 in neurons. PMID- 15361546 TI - Viral macrophage inflammatory protein-II and fractalkine (CX3CL1) chimeras identify molecular determinants of affinity, efficacy, and selectivity at CX3CR1. AB - Fractalkine (FKN/CX3CL1) is a cell surface-expressed chemokine involved in many aspects of leukocyte trafficking and activation. The various structural domains of FKN play distinct roles in its ability to bind and activate its receptor, CX3CR1. A human herpesvirus 8-encoded chemokine, termed viral macrophage inflammatory protein (vMIP)-II, is structurally similar to FKN; vMIP-II is a nonselective chemokine receptor antagonist (binding multiple chemokine receptors, including CX3CR1). The goal of this study was to identify FKN determinants of selectivity for its receptor and to further refine domains important in affinity and efficacy at CX3CR1. Chimeric and insertional mutagenesis was used to generate mutants of both vMIP-II and FKN, and the expressed proteins were evaluated for chemokine receptor binding affinities and efficacy at CX3CR1. Modification of the intervening amino acids between the first two conserved cysteine residues of FKN or vMIP-II indicated a role of the X3 bulge of FKN in affinity and selectivity for CX3CR1. Substitution of the vMIP-II N terminus with that of FKN created an agonist that was just as potent and efficacious as FKN for binding and stimulating CX3CR1, whereas replacement of the FKN N terminus with the cognate domain of vMIP-II disrupted the ability of FKN to bind CX3CR1. Furthermore, the entire N terminus of FKN was necessary for the high-affinity and full agonist properties of FKN at CX3CR1. These results refine the pharmacophore for chemokine binding to and activation of CX3CR1 and demonstrate the usefulness of modified virally encoded chemokines as templates for the development of selective chemokine receptor antagonists. PMID- 15361547 TI - Suppression of pathogenicity of Porphyromonas gingivalis by newly developed gingipain inhibitors. AB - Arg-gingipain (Rgp) and Lys-gingipain (Kgp) are cysteine proteinases produced by Porphyromonas gingivalis, a major etiological bacterium of periodontal diseases. Here we show a series of small peptide analogs able to inhibit either Rgp or Kgp, which are synthesized on the basis of the cleavage site specificity of human salivary histatins by each enzyme. Among this series of compounds, carbobenzoxy Lys-Arg-CO-Lys-N-(CH2)2 (KYT-1) and carbobenzoxy-Glu(NHN(CH3)Ph)-Lys-CO-NHCH2Ph (KYT-36) were found to be the most potent inhibitors of Rgp and Kgp, respectively, with Ki values of 10(-11) to 10(-10) M order. Both inhibitors exhibited slight or no inhibition on mammalian proteinases such as trypsin and cathepsins B, L, and H. All of the virulence induced by the culture supernatant of P. gingivalis tested, including the degradation of various host proteins such as human type I collagen, immunoglobulins, fibronectin, and fibrinogen, disruption of the bactericidal activity of polymorphonuclear leukocytes, and enhancement of the vascular permeability, were strongly inhibited by a combined action of both inhibitors. The functions essential for the bacterium to grow and survive in the periodontal pocket, such as coaggregation and acquisition of amino acids, were also strongly inhibited by the combined action of both inhibitors. The disruption of the adhesion and viability of human fibroblasts and hemagglutination by the organism were strongly suppressed by a single use of KYT 1. These results thus indicate that the newly developed KYT-1 and KYT-36 both should provide a broader application in studies of this important class of enzymes and facilitate the development of new approaches to periodontal diseases. PMID- 15361548 TI - Protein kinase C activation enhances morphine-induced rapid desensitization of mu opioid receptors in mature rat locus ceruleus neurons. AB - Recent studies have shown that morphine, in contrast to other agonists at the mu opioid receptor, causes very little rapid mu-opioid receptor desensitization or internalization in adult rat mammalian neurons, raising important questions about how morphine tolerance is induced. Here we show that morphine can indeed cause marked rapid desensitization of mu-opioid receptors in mature rat locus ceruleus neurons when protein kinase C is also activated. Thus, activation of Gq-coupled M3 muscarinic receptors or application of a phorbol ester enhanced the desensitization of the mu-opioid receptor-evoked potassium current in rat locus ceruleus neurons. The enhancement of desensitization was reversible by the protein kinase C inhibitors chelerythrine and 2-[1-(3-dimethylaminopropyl)-1H indol-3-yl]-3-(1H-indol-3-yl)-maleimide (GF109203X) and resulted from an effect at the level of the mu-opioid receptor rather than the potassium channel. This is the first finding that morphine can induce rapid mu-opioid receptor desensitization in adult rat neurons, and because reduced protein kinase C activity in vivo attenuates morphine tolerance, we propose that G-protein coupled receptor cross-talk and the level of protein kinase C activity may play critical roles in the desensitization of the mu-opioid receptor and could underlie the development of morphine tolerance. PMID- 15361549 TI - Low-affinity neurotensin receptor (NTS2) signaling: internalization-dependent activation of extracellular signal-regulated kinases 1/2. AB - The role and signaling properties of the low-affinity neurotensin receptor (NTS2) are still controversial. In particular, it is unclear whether neurotensin acts as an agonist, inverse agonist, or antagonist at this site. In view of the growing evidence for a role of NTS2 in antinociception, the elucidation of the pharmacological and coupling properties of this receptor is particularly critical. In the present study, we demonstrate that in Chinese hamster ovary (CHO) cells expressing the rat NTS2 receptor, neurotensin (NT), levocabastine, neuromedin N, and the high-affinity NT receptor antagonist SR48692 [2-[[1-(-7 chloroquinolin-4-yl)-5-(2,6-dimethoxyphenyl)-1H-pyrazole-3 carbonyl]amino]adamantane-2-carboxylic acid] all bind to and activate the NTS2 receptor. This activation is followed by ligand-induced internalization of receptor-ligand complexes, as evidenced by confocal microscopy using a fluorescent NT analog. All compounds tested produced a rapid and sustained activation of extracellular signal-regulated kinases 1/2 (ERK1/2) but were without specific effect on Ca(2+) mobilization. The agonist-induced activation of ERK1/2 was completely abolished by preincubation of the cells with the endocytosis inhibitors phenylarsine oxide and monodansylcadaverine as well as overexpression of a dominant-negative mutant of dynamin 1 (DynK44A), indicating that receptor internalization was required for ERK1/2 activation. NTS2-induced activation of ERK1/2 was not species-specific, because the same agonistic effects of NT and analogs were observed in CHO cells transfected with the human NTS2 receptor. In conclusion, this study demonstrates that NTS2 is a bona fide NT receptor and that activation of this receptor by NT or NT analogs results in an internalization-dependent activation of the ERK1/2 signaling cascade. PMID- 15361550 TI - Up-regulation of cyclooxygenase-2 expression is involved in R(+)-methanandamide induced apoptotic death of human neuroglioma cells. AB - Cannabinoids have been implicated in the reduction of glioma growth. The present study investigated a possible relationship between the recently shown induction of cyclooxygenase (COX)-2 expression by the endocannabinoid analog R(+)methanandamide [R(+)-MA] and its effect on the viability of H4 human neuroglioma cells. Incubation with R(+)-MA for up to 72 h decreased the cellular viability and enhanced accumulation of cytoplasmic DNA fragments in a time dependent manner. Suppression of R(+)-MA-induced prostaglandin (PG) E2 synthesis with the selective COX-2 inhibitor celecoxib (0.01-1 microM) or inhibition of COX 2 expression by COX-2-silencing small-interfering RNA was accompanied by inhibition of R(+)-MA-mediated DNA fragmentation and cell death. In contrast, the selective COX-1 inhibitor SC-560 was inactive in this respect. Cells were also protected from apoptotic cell death by other COX-2 inhibitors (NS-398 [[N-[2 (cyclohexyloxy)-4-nitrophenyl]-methanesulfonamide]] and diclofenac) and by the ceramide synthase inhibitor fumonisin B1, which interferes with COX-2 expression by R(+)-MA. Moreover, the proapoptotic action of R(+)-MA was mimicked by the major COX-2 product PGE2. Apoptosis and cell death by R(+)-MA were not affected by antagonists of cannabinoid receptors (CB1, CB2) and vanilloid receptor 1. In further experiments, celecoxib was demonstrated to suppress apoptotic cell death elicited by anandamide, which is structurally similar to R(+)-MA. As a whole, this study defines COX-2 as a hitherto unknown target by which a cannabinoid induces apoptotic death of glioma cells. Furthermore, our data show that pharmacological concentrations of celecoxib may interfere with the proapoptotic action of R(+)-MA and anandamide, suggesting that cotreatment with COX-2 inhibitors could diminish glioma regression induced by these compounds. PMID- 15361551 TI - Enhancement of cardiac L-type Ca2+ currents in transgenic mice with cardiac specific overexpression of CYP2J2. AB - CYP2J2 is abundant in cardiomyocytes and is involved in the metabolism of arachidonic acid (AA) to epoxyeicosatrienoic acids (EETs), which affect multiple cell functions. In this study, we investigated the effect of overexpression of CYP2J2 on cardiac L-type Ca2+ currents (ICa) in adult transgenic mice. Cardiac specific overexpression of CYP2J2 was achieved using the alpha-myosin heavy chain promoter. ICa was recorded from isolated ventricular cardiomyocytes. Compared with the wild-type cardiomyocytes (n = 60), the density of ICa was significantly increased by 40 +/- 9% in the CYP2J2 transgenic cardiomyocytes (n = 71; P < 0.001). N-Methylsulfonyl-6-(2-proparglyloxyphenyl)hexanamide (MS-PPOH), a specific inhibitor of EET biosynthesis, and clotrimazole, a cytochrome P450 inhibitor, significantly reduced ICa in both wild-type and transgenic cardiomyocytes; however, MS-PPOH inhibited ICa to a greater extent in the CYP2J2 transgenic cells (n = 10) than in the wild-type cells (n = 10; P < 0.01). Addition of 11,12-EET significantly restored ICa in MS-PPOH-treated cells. Intracellular dialysis with either of two inhibitory monoclonal antibodies against CYP2J2 significantly reduced ICa in both wild-type and transgenic mice. Membrane-permeable 8-bromo-cAMP and the beta-adrenergic agonist isoproterenol significantly reversed the monoclonal antibody-induced inhibition of ICa. In addition, the total protein level of the alpha1 subunit of the Cav1.2 L-type Ca2+ channel was not altered in CYP2J2 transgenic hearts, but the phosphorylated portion was markedly increased. In conclusion, overexpression of CYP2J2 increases ICa in CYP2J2 transgenic cardiomyocytes via a mechanism that involves cAMP protein kinase A-dependent phosphorylation of the L-type Ca2+ channel. PMID- 15361552 TI - Labour is associated with increased expression of type-IIA secretory phospholipase A2 but not type-IV cytosolic phospholipase A2 in human myometrium. AB - Human labour is associated with increased prostaglandin synthesis within the uterus. The aim of this study was to examine the expression of the type-IV cytosolic phospholipase A2 (cPLA2-IV) and the type IIA secretory phospholipase A2 (sPLA2-IIA) in myometrium in association with labour onset at term and preterm deliveries. These enzymes are important for the release of the prostaglandin precursor, arachidonic acid, from phospholipid membrane stores. RT-PCR was used to determine differences in gene expression between non-labour and labour groups. Expression of sPLA2-IIA in human myometrium was significantly increased with pregnancy, and with labour, both at term and preterm. Expression of cPLA2-IV in myometrium was not significantly altered with respect to pregnancy or labour. Immunohistochemical analysis demonstrated differences in the spatial localization of cPLA2-IV and sPLA2-IIA protein in upper and lower segment myometrium. cPLA2-IV was predominantly in vascular endothelial cells, while sPLA2-IIA was observed in vascular, endothelial and smooth muscle cells. In addition, sPLA2-IIA showed a distinct nuclear or perinuclear localization in myometrial smooth muscle cells of the lower segment. We postulate that the increased expression of sPLA2-IIA rather than cPLA2-IV in the myometrium may play a role in the onset and/or maintenance of human parturition. PMID- 15361553 TI - Role of volume-stimulated osmolyte and anion channels in volume regulation by mammalian sperm. AB - The ability to maintain cellular volume is an important general physiological function. Swelling induced by hypotonic stress results in the opening of channels, through which ions exit with accompanying water loss (regulatory volume decrease, RVD). RVD has been shown to occur in mammalian sperm, primarily through the opening of quinine-sensitive potassium channels. However, as yet, direct evidence for the participation of anion channels in sperm RVD has been lacking. The chloride channel type ClC-3 is believed to be involved in RVD in other cell types. Using electronic cell sizing for cell volume measurement, the following results were obtained. (i) The anion channel blockers 5-nitro-2-(3 phenylpropylamino)-benzoic acid (NPPB), tamoxifen and 4,4'-diisothiocyanostilbene 2,2'-disulphonic acid (DIDS) increased hypotonic swelling in concentration dependent fashion, whereas verapamil (P-glycoprotein inhibitor) had little effect. The most potent, NPPB and DIDS, blocked RVD without affecting cell membrane integrity at effective concentrations. (ii) When gramicidin was included to dissipate Na+/K+ gradients, major secondary swelling was observed under hypotonic conditions. This secondary swelling could be reduced by NPPB, and suppressed completely by replacing chloride in the medium with sulphate, an ion which does not pass through chloride channels. It was deduced that the initial hypotonic swelling activated an anion channel through which chloride ions could then enter freely down a concentration gradient, owing to the lack of a counter gradient of potassium. (iii) Taurine, an osmolyte often involved in RVD, does not appear to play a role in sperm RVD because lengthy preincubation with taurine did not alter sperm RVD response. Our observations provide direct evidence that a chloride channel (possibly ClC-3) is involved in the process of volume regulation in mammalian sperm. PMID- 15361554 TI - Rapid prenatal diagnosis of common chromosome aneuploidies by QF-PCR. Assessment on 18,000 consecutive clinical samples. AB - The quantitative fluorescent PCR (QF-PCR) assay, introduced during the last few years, allows prenatal diagnoses of common chromosome aneuploidies in a few hours after sampling. We report the first assessment of QF-PCR performed on a large cohort of 18,000 consecutive clinical specimens analysed in two different Centres. All samples were analysed by QF-PCR using several selected STR markers together with amelogenin and, occasionally, SRY for fetal sexing. Results were compared with those obtained by conventional cytogenetic analysis. In 17,129 tests, normal fetuses were detected by QF-PCR. No false positives were observed. All 732 cases of trisomy 21, 18, 13, triploidies, double trisomies as well as all but one fetuses with X and Y aneuploidies were correctly diagnosed. Chromosome mosaicism could also be suspected in several samples. In some cases of in vitro culture failures, QF-PCR was the only evidence of fetal X, Y, 21, 18 and 13 chromosome complement. QF-PCR proved to be efficient and reliable in detecting major numerical chromosome disorders. The main advantages of the molecular assay are its very low cost, speed and automation enabling a single operator to perform up to 40 assays per day. QF-PCR relieves anxiety of most parents within 24 h from sampling and accelerates therapeutic interventions in the case of an abnormal result. In countries where large scale conventional cytogenetics is hampered by its high cost and lack of technical expertise, QF-PCR may be used as the only prenatal diagnostic test. PMID- 15361555 TI - Inhibin, activin, follistatin, activin receptors and beta-glycan gene expression in the villous tissue of miscarriage patients. AB - Maternal circulating levels of inhibin A are significantly lower in patients with clinical symptoms of miscarriage. The objective of this study was to quantify relative expression of inhibin alpha, inhibin/activin betaA, betaB, betaC, follistatin, activin receptors and beta-glycan genes and content of inhibin A, activin A and follistatin protein in villous tissue of first trimester miscarriages and gestation-matched normal pregnancies. Twelve women with clinical symptoms of miscarriage were matched with 12 normal pregnancies for gestational age. Total RNA was isolated from placental samples. Complementary DNA produced by reverse transcription was used in the real-time PCR to quantify the expression of the genes. The ratio between the target and rRNA 18S was calculated to provide relative gene expression. Villous tissue homogenates were used for the determination of the content of inhibin A, activin A and follistatin protein. Maternal serum was assayed for inhibin A, activin A and follistatin. All villous samples expressed inhibin alpha, inhibin/activin betaA, betaB, betaC, follistatin, activin receptors (ACTRIA, ACTRIB, ACTRIIA, ACTRIIB) and beta-glycan genes. There was no significant difference in the relative expression of these genes between the groups. Villous content of inhibin A, activin A and follistatin were also not different between the two groups. Maternal serum levels of inhibin A were significantly lower in the miscarriage group compared to the controls. The decreased maternal levels of inhibin A in miscarriage patients could be due to a decrease in placental mass prior to embryonic demise. This finding also confirms that the trophoblast is the major source of inhibin A after the luteo-placental shift in early pregnancy. PMID- 15361556 TI - 2004 Dr. Gary J. Becker Young Investigator Award: Relative thermosensitivity of cytotoxic drugs used in transcatheter arterial chemoembolization. AB - PURPOSE: Large hepatocellular carcinoma tumors are being treated increasingly with a combination of transcatheter arterial chemoembolization (TACE) and radiofrequency (RF) ablation. However, the high temperatures reached during RF ablation may reduce the cytotoxic effects of antineoplastic agents, but this has not been studied. Therefore, in the present study, the relative thermosensitivity of cytotoxic drugs commonly used in TACE was studied. MATERIALS AND METHODS: The relative cytotoxic effects of cisplatin, doxorubicin HCl, and mitomycin on the growth of human colon HT29 and lung A549 adenocarcinoma cells before and after heating each drug in solution was determined from the standpoints of different durations of exposure (15, 30, 60, 90, and 120 minutes) at a fixed temperature (120 degrees C) and exposure to different temperatures (60 degrees C, 80 degrees C, 100 degrees C, and 120 degrees C) for a fixed period of time (2 hours). After 72 hours of exposure of the cells to each drug, relative cell growth inhibition was assessed by MTT assay, and 50% inhibitory concentration (IC(50)) values were calculated for each cytotoxic agent. Finally, the heat-dependent degradation of mitomycin and doxorubicin was analyzed with use of tandem electrospray mass spectrometry. RESULTS: The relative cytotoxic activities (shown by cell growth inhibition and IC(50) values) of cisplatin, doxorubicin, and mitomycin heated to 120 degrees C for 2 hours decreased by factors of 1.35 (range, 1-1.75), 9.5 (range, 8.5-10.5), and 7.05 (range, 3.5-12), respectively. The cytotoxic activities of doxorubicin and mitomycin continued to decrease with incremental increases in temperature. Similarly, with incremental increases in the duration of exposure to heat, the cytotoxic activities of doxorubicin and mitomycin decreased. Mass spectrometric analysis of residual drug content showed that a 2 hour exposure to a temperature of 120 degrees C caused doxorubicin and mitomycin to degrade by 95% and 84%, respectively. CONCLUSIONS: The cytotoxicity of cisplatin is not affected by heat. The cytotoxicities of doxorubicin and mitomycin are reduced by high temperature and duration of exposure to heat. Although degradation of cytotoxicity starts at 60 degrees C and after 30 minutes of exposure to heat, statistically significant changes are encountered at 100 degrees C and after 90 minutes of exposure. PMID- 15361557 TI - Comparing the SMART stent with the Wallstent iliac endoprosthesis: reading between the lines. PMID- 15361558 TI - The Nitinol SMART stent vs Wallstent for suboptimal iliac artery angioplasty: CRISP-US trial results. AB - PURPOSE: The Cordis Randomized Iliac Stent Project-US (CRISP-US) trial evaluated, with an equivalence design, the performance of the shape memory alloy recoverable technology (SMART) nitinol self-expanding stent and the stainless steel Wallstent for treating iliac artery disease after suboptimal percutaneous transluminal angioplasty (PTA). MATERIALS AND METHODS: This multicenter, prospective, randomized trial comprised 203 patients with chronic limb ischemia who received either the SMART stent (n = 102) or the Wallstent (n = 101) after suboptimal PTA. The primary equivalence end point was a composite of 9-month restenosis, 30-day death, and 9-month target vessel revascularization. Functional, clinical, and hemodynamic assessments were made at hospital discharge and at 1, 6, 9, and 12 months. RESULTS: The 9-month composite end point rate was equivalent for the SMART stent and Wallstent (6.9% vs 5.9%), with low rates of restenosis (3.5% vs 2.7%), death (2.0% vs 0.0%), and revascularization (2.0% vs 4.0%) in the two groups. Primary patency at 12 months was 94.7% and 91.1% with the SMART stent and Wallstent, respectively. Functional and hemodynamic improvement was also comparable between the groups. The acute procedural success rate was higher in the SMART stent group (98.2% vs 87.5%; P =.002). The frequency of major adverse events was similar at 1 year (4.9% vs 5.9%). CONCLUSIONS: The performance of the SMART stent was equivalent to that of the Wallstent for treating iliac artery stenosis. The design characteristics of the SMART stent may contribute to greater procedural success and more accurate stent deployment. PMID- 15361559 TI - Radiation doses in interventional radiology procedures: the RAD-IR Study. Part III: Dosimetric performance of the interventional fluoroscopy units. AB - PURPOSE: To present the physics data supporting the validity of the clinical dose data from the RAD-IR study and to document the performance of dosimetry components of these systems over time. MATERIALS AND METHODS: Sites at seven academic medical centers in the United States prospectively contributed data for each of 12 fluoroscopic units. All units were compatible with International Electrotechnical Commission (IEC) standard 60601-2-43. Comprehensive evaluations and periodic consistency checks were performed to verify the performance of each unit's dosimeter. Comprehensive evaluations compared system performance against calibrated ionization chambers under nine combinations of operating conditions. Consistency checks provided more frequent dosimetry data, with use of each unit's built-in dosimetry equipment and a standard water phantom. RESULTS: During the 3 year study, data were collected for 48 comprehensive evaluations and 581 consistency checks. For the comprehensive evaluations, the mean (95% confidence interval range) ratio of system to external measurements was 1.03 (1.00-1.05) for fluoroscopy and 0.93 (0.90-0.96) for acquisition. The expected ratio was 0.93 for both. For consistency checks, the values were 1.00 (0.98-1.02) for fluoroscopy and 1.00 (0.98-1.02) for acquisition. Each system was compared across time to its own mean value. Overall uncertainty was estimated by adding the standard deviations of the comprehensive and consistency measurements in quadrature. The authors estimate that the overall error in clinical cumulative dose measurements reported in RAD-IR is 24%. CONCLUSION: Dosimetric accuracy was well within the tolerances established by IEC standard 60601-2-43. The clinical dose data reported in the RAD-IR study are valid. PMID- 15361560 TI - Intravascular ultrasound-guided direct intrahepatic portacaval shunt: midterm follow-up. AB - PURPOSE: To describe the midterm patency and clinical results of the intravascular ultrasound (US)-guided direct intrahepatic portacaval shunt (DIPS) procedure for the treatment of intractable ascites and variceal bleeding. MATERIALS AND METHODS: From February 23, 1999, to December 18, 2002, inferior vena cava (IVC)-to-portal vein (PV) shunts were created in 40 patients for ascites (n = 35) and bleeding (n = 5). Intravascular US was used to guide direct puncture from the IVC to the PV. The shunts were completed with the use of single (n = 15) or overlapping (n = 25) polytetrafluoroethylene (PTFE)-covered Palmaz stents. These stent-grafts were deployed primarily at diameters of 8 mm. The diameter of the shunt was increased in three cases to achieve a target portosystemic gradient (PSG) of 15 mm Hg or lower. All patients were followed clinically and with portography with manometry. RESULTS: All DIPSs were created successfully. Mean PSGs were reduced from 23 mm Hg before DIPS creation to 9 mm afterward. During the follow-up period of a maximum of 38 months, 22 of 40 patients died or underwent liver transplantation (mean follow-up, 9 months; median, 6 months). At the time of this report, 18 of 40 patients remain living after follow-up ranging in duration from 8 to 38 months (mean, 22 months; median, 16.5 months). During the follow-up period, there was one stent-graft occlusion and three stenoses. These four patients were successfully treated by additional stent-graft placement. In addition, two patients developed IVC stenosis cephalad to the DIPS, which required IVC stent placement. The primary patency rates by Kaplan-Meier analysis were 100% at 6 months and 75% (95% CI, 53%-97%) at 12 months. CONCLUSION: Intravascular US-guided direct IVC-to-PV shunts may be created successfully with minimal complications. Primary patency of the shunt is greater than that with conventional TIPS with a bare wire stent and appears equal to that with TIPS with a PTFE-covered stent-graft. PMID- 15361561 TI - Embolization of peripheral vascular malformations with ethylene vinyl alcohol copolymer (Onyx). AB - PURPOSE: To demonstrate the feasibility and preliminary efficacy of endovascular embolization of peripheral congenital vascular malformations (VMs) with use of a nonadhesive liquid embolic agent, Onyx. MATERIALS AND METHODS: Nine patients with a mean age of 20.8 years had local low-flow (n = 4), local high-flow (n = 3), or diffuse high-flow (n = 2) VMs located in the upper or lower extremities. In all patients, endovascular embolization was performed via the superselective catheterization of arterial feeders of VMs with use of microcatheters in a coaxial technique. A total of 15 embolization procedures were performed with Onyx, which was composed of 6%, 8%, or 20% ethylene vinyl alcohol copolymer dissolved in dimethyl sulfoxide. RESULTS: In two of four patients with local low flow VMs, the lesions were embolized completely. In the other two patients with local low-flow VMs, embolizations were incomplete. The remaining five high-flow lesions of local (n = 3) or diffuse (n = 2) types were also embolized incompletely. In all patients with local low-flow VMs and in one patient with a local high-flow VM, clinical signs and symptoms were resolved significantly. Other patients did show clinical benefit from embolization to varying degrees. CONCLUSION: In our experience in a limited number of cases, Onyx promises and provides important advantages over conventional embolic agents in the endovascular transcatheter embolization of congenital peripheral VMs. However, as with other embolic agents, it is far from perfect. PMID- 15361562 TI - Aortoiliac gadolinium-enhanced CT angiography: improved results with a 16 detector row scanner compared with a four-detector row scanner. AB - PURPOSE: To assess the level of vascular enhancement of gadolinium-enhanced aortoiliac computed tomographic (CT) angiography with a 16-detector row CT scanner and to compare it with the results of previous similar studies that used four-detector row CT units. MATERIALS AND METHODS: Eleven gadolinium-enhanced CT angiograms were obtained in 10 consecutive patients with contraindication to iodinated contrast medium with use of a 16-detector row CT scanner. In the region of interest, attenuation measurements (in HU) were obtained from the proximal abdominal aorta to the common femoral arteries during unenhanced, gadolinium enhanced, and delayed acquisitions. The results were compared to those in the 15 consecutive patients who most recently had similar examinations performed on a four-detector row CT unit. Phantom studies with diluted gadolinium were conducted to compare attenuation between CT units. RESULTS: On four-detector row CT, throughout the scan length, mean enhancement values were 53.8 HU +/- 5.3 and 15.0 HU +/- 2.6 for gadolinium-enhanced and delayed series, respectively. For the 16 detector row CT unit, they were 76.1 HU +/- 3.4 and 21.3 HU +/- 1.3, respectively. As a result of a shorter scan time and a more optimal start time, the 16-detector row CT unit provided significantly greater and more consistent enhancement throughout the scan length compared with the four-detector row CT unit (P =.0106). Similar structures had significantly greater enhancement when 120 kV was applied instead of 140 kV (P =.0495) CONCLUSION: The 16-detector row CT scanner improved gadolinium-enhanced CT angiography results compared with the four-detector row CT unit. PMID- 15361563 TI - CT-guided percutaneous lung biopsy in children. AB - PURPOSE: To describe techniques and evaluate outcomes of computed tomography (CT) guided percutaneous lung biopsy in children. MATERIALS AND METHODS: Between April 1992 and June 2003, 64 patients (32 male, 32 female) with a mean age of 10.8 years (0.6-20 years) were referred for 75 lung biopsies. Most biopsies were performed for suspected malignancy (n = 24; 32%) or to distinguish posttransplantation lymphoproliferative disorder from fungal infection in immunocompromised patients (n = 17; 23%). All children referred to the pediatric interventionalists in two children's hospitals for CT-guided biopsy of parenchymal or pleural-based lesions in the thorax were studied. Prospectively gathered procedural data were reviewed for medical history and indications for procedure, admission status, type of anesthesia, technical approach (core vs aspiration biopsy), procedural modifications, lesion size, number of passes required, and immediate complications. Medical records were retrospectively reviewed for diagnostic outcome, impact on patient management, and delayed complications. RESULTS: Procedures were performed under deep sedation whenever possible (n = 61; 81%) with use of a coaxial core biopsy technique (n = 56; 75%), a fine needle aspiration biopsy technique (n = 15; 20%), or both (n = 4; 5%). Mean lesion diameters were 2.5 cm (range, 1-10 cm) in the core biopsy group and 1.0 cm (range, 0.5-1.7 cm) in the aspiration biopsy group. Sixty-four biopsies (85%) were diagnostic. There was one major complication (1.3%), a tension pneumothorax treated with intraprocedural placement of a chest tube. CONCLUSION: Percutaneous CT-guided lung biopsy is a safe and accurate diagnostic procedure in children that obviates open surgical biopsy in most patients. PMID- 15361564 TI - Multislice CT virtual intravascular endoscopy for abdominal aortic aneurysm stent grafts. AB - PURPOSE: To investigate the optimal scanning protocols of multislice computed tomographic (CT) angiography in aortic stent graft placement observed on virtual intravascular endoscopy (VIE). MATERIALS AND METHODS: A human aorta phantom was built with a commercial aortic stent graft placed in situ. The phantom was housed in a perspex container and filled with contrast medium with CT attenuation similar to that used in the patient's abdominal scanning. CT was performed with a four-slice multislice CT scanner with section thickness of 1.3 mm, 2 mm, and 3.2 mm, pitch of 0.75, 1, and 1.5, and reconstruction interval of 50% overlap and no overlap. Stair-step artifacts were quantified by measuring the SD of signal intensity on surfaced shaded VIE images in three different locations: superior mesenteric artery (SMA), renal ostium, and aortic aneurysm. RESULTS: Image quality of VIE images was independent of section thickness and pitch values in the level of renal ostium and aortic aneurysm (P >.05), whereas it was determined by the section thickness and pitch in the level of SMA (P <.05). Aortic branch origin became irregular or distorted when section thickness increased to 3.2 mm or pitch reached 1.5. CONCLUSION: A scanning protocol of section thickness of 2 mm, pitch 1, and reconstruction interval of 1 mm is recommended in aortic stent graft placement because it allows fewer stair-step artifacts and better visualization of the aortic stent wires observed on VIE. PMID- 15361565 TI - Endovascular aortic aneurysm repair with stent-grafts: experimental models can reproduce endoleaks. AB - PURPOSE: To develop canine aneurysm models that can reproduce type II endoleaks after endovascular aneurysm repair (EVAR) with stent-grafts. MATERIALS AND METHODS: A fusiform infrarenal abdominal aortic aneurysm model (AAA) was surgically created with a jugular vein patch, while preserving collateral vessels (n = 3). To allow comparative studies within the same animal, a bilateral iliac aneurysm model was also constructed with venous patches and surgical re implantation of the sacroiliac trunk (n = 3). Stent-grafts were implanted by femoral approach at least 2 months later in both aortic and iliac models. Follow up imaging was performed by Doppler ultrasound (US) and angiography until animals were killed 3 months after EVAR. RESULTS: Angiography revealed immediate type II leaks in all cases. Leaks were still present at autopsy 3 months after EVAR in all cases, and were revealed at pre-death angiography in all but one case. At autopsy, leaks were characterized by the presence of large endothelialized channels that formed within the thrombus between the stent-graft and the aneurysmal wall. CONCLUSION: As shown in this pilot study, persistent type II leaks after EVAR can be reproduced in aortic and iliac animal models. The iliac model can be created bilaterally in the same animal, thus allowing for comparative evaluation of different therapies. These models could be used to better understand the mechanisms of endoleak, and to assess future developments aimed to improve the outcomes after EVAR. PMID- 15361566 TI - Intraarterial contrast material-enhanced magnetic resonance angiography of the aortoiliac system. AB - In a period of 4 months, 10 patients were examined prospectively with intraarterial magnetic resonance (MR) angiography after digital subtraction angiography. Intraarterial MR angiography was performed with use of a 1.5-T MR imaging system. Contrast agent (gadodiamide) was injected with a conventional angiography catheter placed in the proximal abdominal aorta. The increase in vascular signal intensity was determined and the diagnostic value of the MR angiograms was scored according to a five-point scale by four investigators. The MR angiograms were judged good to excellent, and all were scored as diagnostic. In conclusion, this study shows that intraarterial MR angiography is feasible. Intraarterial MR angiography is appropriate to support MR-guided vascular intervention. PMID- 15361567 TI - Guide wire perforation leading to fatal perirenal hemorrhage from transcortical collaterals after renal artery stent placement. AB - After successful renal artery angioplasty and stent placement, a patient in a fully anticoagulated state developed hypotension and flank pain. Review of the intraprocedural angiogram demonstrated transcortical position of the guide wire. Computed tomography of the abdomen revealed a large perinephric hematoma. Although only a single renal artery branch was accessed with the guide wire during stent placement, subsequent emergent angiography revealed extravasation from multiple capsular branches. Renal artery embolization failed to control the hemorrhage. The patient's course rapidly deteriorated and he ultimately died. The unique angiographic finding and proposed mechanism of this fatal complication are described and discussed. PMID- 15361568 TI - Endovascular treatment of congenital portal vein fistulas with the Amplatzer occlusion device. AB - Two patients with symptomatic congenital portal vein (PV) fistulas (one case of PV-right hepatic vein fistula and one case of PV-inferior vena cava fistula) causing hepatopulmonary syndrome (hepatic dysfunction, lung vascular alteration in the form of arteriovenous shunts, and hypoxemia) were successfully treated with the Amplatzer patent ductus arteriosus (PDA) occlusion device. This device was chosen in light of the anatomic characteristics of the vascular malformations and the specific properties of the prosthesis, especially those related to relocation and retrievability when not properly positioned. Embolization with an Amplatzer PDA occlusion device should be considered as a useful therapeutic alternative in the treatment of congenital portal fistulas that can obviate complex surgeries. PMID- 15361569 TI - Inferior vena cava thrombosis after transjugular intrahepatic portosystemic shunt revision with a covered stent. AB - A 42-year-old woman who had undergone multiple revisions of a bare-stent transjugular intrahepatic portosystemic shunt was treated for in-stent stenosis by insertion of a polytetrafluoroethylene (PTFE)-covered stent. Immediately after revision with the covered stent, she developed inferior vena cava (IVC) thrombosis. The potential causes and implications of this complication are discussed. PMID- 15361570 TI - Percutaneous cystic duct stent placement for the treatment of acute cholecystitis resulting from common bile duct stent placement for malignant obstruction. AB - Common bile duct (CBD) stent placement to relieve malignant biliary obstruction can occasionally cause cystic duct obstruction and acute cholecystitis. Cholecystostomy tube placement is often performed in patients with limited life expectancy but can have a significant impact on quality of life. To allow cholecystostomy tube removal, percutaneous metallic stent placement was performed across the cystic duct via the tube tract in such a patient. The procedure included traversal across the previously placed CBD stent. At 5-month follow-up, the patient remained symptom-free. In select patients who develop acute cholecystitis after CBD stent placement for malignant obstruction, percutaneous stent placement across the cystic duct may be considered a treatment option. PMID- 15361571 TI - Results of temporary placement of covered retrievable expandable nitinol stents for tuberculous bronchial strictures. AB - Covered retrievable expandable nitinol stents were successfully placed in nine patients with tuberculous bronchial strictures refractory to balloon dilation. Complications included stent migration (n = 1) and tissue hyperplasia at the proximal portion of the stent (n = 2). During the follow-up period (5-52 months), stricture recurred in three of five patients (60%) after temporary stent placement for 2 months, whereas stricture did not recur in the other four patients after temporary stent placement for 6 months. Temporary placement of covered retrievable stents appears to be an effective method for the treatment of patients with tuberculous bronchial strictures refractory to balloon dilation. PMID- 15361572 TI - SIR 2004 film panel case: Systemic artery aneurysms in Kawasaki disease. PMID- 15361573 TI - SIR 2004 film panel case: Gangrene caused by homocystinuria. PMID- 15361574 TI - SIR 2004 film panel case: Primary hyperoxaluria type I mimicking arterial vasculitis--a lethal disease. PMID- 15361575 TI - SIR 2004 film panel case: Multicentric granular cell tumor with biliary, subcutaneous, and breast involvement. PMID- 15361576 TI - Reconstruction of the basilar tip with T stent configuration for treatment of a wide-neck aneurysm. PMID- 15361577 TI - Loss of vision as a complication of nasolacrimal stent placement. PMID- 15361578 TI - Domain swapping is a consequence of minimal frustration. AB - The same energy landscape principles associated with the folding of proteins into their monomeric conformations should also describe how these proteins oligomerize into domain-swapped conformations. We tested this hypothesis by using a simplified model for the epidermal growth factor receptor pathway substrate 8 src homology 3 domain protein, both of whose monomeric and domain-swapped structures have been solved. The model, which we call the symmetrized Go-type model, incorporates only information regarding the monomeric conformation in an energy function for the dimer to predict the domain-swapped conformation. A striking preference for the correct domain-swapped structure was observed, indicating that overall monomer topology is a main determinant of the structure of domain-swapped dimers. Furthermore, we explore the free energy surface for domain swapping by using our model to characterize the mechanism of oligomerization. PMID- 15361579 TI - 129Xe on Ir(111): NMR study of xenon on a metal single crystal surface. AB - NMR experiments of (129)Xe adsorbed on an iridium single crystal surface are reported. Very high nuclear polarization (P(z) approximately 0.7) makes the experiment possible. A coverage of less then one monolayer is investigated on the Ir(111) surface with an area of 0.8 cm(2). The observed resonance line shifts are very large and highly anisotropic. We find sigma(iso) = 1,032 +/- 11 ppm and sigma(an) = 291 +/- 33 ppm, which are far above the typical range of physisorption. The highly ordered substrate leads to homogeneous conditions for the xenon atoms, as seen in the narrow linewidth of 20 ppm. Chemical shifts under physisorption conditions are not large enough to totally explain the results. Knight shift can clearly be identified as the cause of the findings. This shift shows the presence of conduction electrons of the metallic substrate at the xenon nucleus and thus the mixing of metallic and atomic states at the Fermi level. Such mixing is in accordance with recent Hartree-Fock and density functional calculations of similar van der Waals adsorption systems. Quantitative comparisons, however, fail completely. The size and ratio of sigma(an) and sigma(iso) are pure ground-state properties in a structurally simple system. They are accessible to theory and provide detailed local information that can serve as a benchmark for theory. PMID- 15361581 TI - A quenchable superhard carbon phase synthesized by cold compression of carbon nanotubes. AB - A quenchable superhard high-pressure carbon phase was synthesized by cold compression of carbon nanotubes. Carbon nanotubes were placed in a diamond anvil cell, and x-ray diffraction measurements were conducted to pressures of approximately 100 GPa. A hexagonal carbon phase was formed at approximately 75 GPa and preserved at room conditions. X-ray and transmission electron microscopy electron diffraction, as well as Raman spectroscopy at ambient conditions, explicitly indicate that this phase is a sp(3)-rich hexagonal carbon polymorph, rather than hexagonal diamond. The cell parameters were refined to a(0) = 2.496(4) A, c(0) = 4.123(8) A, and V(0) = 22.24(7) A (3). There is a significant ratio of defects in this nonhomogeneous sample that contains regions with different stacking faults. In addition to the possibly existing amorphous carbon, an average density was estimated to be 3.6 +/- 0.2 g/cm(3), which is at least compatible to that of diamond (3.52 g/cm(3)). The bulk modulus was determined to be 447 GPa at fixed K' identical with 4, slightly greater than the reported value for diamond of approximately 440-442 GPa. An indented mark, along with radial cracks on the diamond anvils, demonstrates that this hexagonal carbon is a superhard material, at least comparable in hardness to cubic diamond. PMID- 15361580 TI - Dissection of the role of MHC class II A and E genes in autoimmune susceptibility in murine lupus models with intragenic recombination. AB - Systemic lupus erythematosus (SLE) is a multigenic autoimmune disease, and the major histocompatibility complex (MHC) class II polymorphism serves as a key genetic element. In SLE-prone (NZB x NZW)F(1) mice, the MHC H-2(d/z) heterozygosity (H-2(d) of NZB and H-2(z) of NZW) has a strong impact on disease; thus, congenic H-2(d/d) homozygous F(1) mice do not develop severe disease. In this study, we used Ea-deficient intra-H-2 recombination to establish A(d/d) congenic (NZB x NZW)F(1) mice, with or without E molecule expression, and dissected the role of class II A and E molecules. Here we found that A(d/d) homozygous F(1) mice lacking E molecules developed severe SLE similar to that seen in wild-type F1 mice, including lupus nephritis, autoantibody production, and spontaneously occurring T cell activation. Additional evidence revealed that E molecules prevent the disease in a dose-dependent manner; however, the effect is greatly influenced by the haplotype of A molecules, because wild-type H-2(d/z) F(1) mice develop SLE, despite E molecule expression. Studies on the potential of dendritic cells to present a self-antigen chromatin indicated that dendritic cells from wild-type F(1) mice induced a greater response of chromatin-specific T cells than did those from A(d/d) F(1) mice, irrespective of the presence or absence of E molecules, suggesting that the self-antigen presentation is mediated by A, but not by E, molecules. Our mouse models are useful for analyzing the molecular mechanisms by which MHC class II regions regulate the process of autoimmune responses. PMID- 15361582 TI - Gene expression profiling reflects physiological processes in salt acclimation of Synechocystis sp. strain PCC 6803. AB - The kinetics of genome-wide responses of gene expression during the acclimation of cells of Synechocystis sp. PCC 6803 to salt stress were followed by DNA microarray technique and compared to changes in main physiological parameters. During the first 30 min of salt stress, about 240 genes became induced higher than 3-fold, while about 140 genes were repressed. However, most changes in gene expression were only transient and observed among genes for hypothetical proteins. At 24 h after onset of salt stress conditions, the expression of only 39 genes remained significantly enhanced. Among them, many genes that encode proteins essential for salt acclimation were detected, while only a small number of genes for hypothetical proteins remained activated. Following the expression of genes for main functions of the cyanobacterial cell, i.e. PSI, PSII, phycobilisomes, and synthesis of compatible solutes, such as ion homeostasis, distinct kinetic patterns were found. While most of the genes for basal physiological functions were transiently repressed during the 1st h after the onset of salt stress, genes for proteins specifically related to salt acclimation were activated. This gene expression pattern reflects well the changes in main physiological processes in salt-stressed cells, i.e. transient inhibition of photosynthesis and pigment synthesis as well as immediate activation of synthesis of compatible solutes. The results clearly document that following the kinetics of genome-wide expression, profiling can be used to envisage physiological changes in the cyanobacterial cell after certain changes in growth conditions. PMID- 15361583 TI - Divergent potentials for cytoplasmic inheritance within the genus Syringa. A new trait associated with speciogenesis. AB - Epifluorescence microscopic detection of organelle DNA in the mature generative cell is a rapid method for determining the potential for the mode of cytoplasmic inheritance. We used this method to examine 19 of the known 22 to 27 species in the genus Syringa. Organelle DNA was undetectable in seven species, all in the subgenus Syringa, but was detected in the 12 species examined of the subgenera Syringa and Ligustrina. Therefore, species within the genus Syringa display differences in the potential cytoplasmic inheritance. Closer examination revealed that the mature generative cells of the species in which organelle DNA was detected contained both mitochondria and plastids, but cells of the species lacking detectable organelle DNA contained only mitochondria, and the epifluorescent organelle DNA signals from the mature generative cells corresponded to plastid DNA. In addition, semiquantitative analysis was used to demonstrate that, during pollen development, the amount of mitochondrial DNA decreased greatly in the generative cells of the species examined, but the amount of plastid DNA increased remarkably in the species containing plastids in the generative cell. The results suggest that all Syringa species exhibit potential maternal mitochondrial inheritance, and a number of the species exhibit potential biparental plastid inheritance. The difference between the modes of potential plastid inheritance among the species suggests different phylogenies for the species; it also supports recent conclusions of molecular, systematic studies of the Syringa. In addition, the results provide new evidence for the mechanisms of maternal mitochondrial inheritance in angiosperms. PMID- 15361585 TI - Ethnic differences in intimate partner violence in the U.S. general population: the role of alcohol use and socioeconomic status. AB - This article reviews cross-sectional and longitudinal research on ethnic differences related to intimate partner violence (IPV) in the U.S. general population and the role of alcohol use and socioeconomic characteristics. Evidence indicates that significant ethnic differences exist in the prevalence of IPV. Although ethnic minorities report higher rates of IPV, differences in crude rates are reduced after controlling for socioeconomic circumstances and alcohol use. However, Black couples appear to be at greater risk of IPV than their White or Hispanic counterparts even after controlling for such risk factors. Overall, socioeconomic characteristics,demographic characteristics, and alcohol use appear to play important roles in the occurrence of IPV. These findings suggests that IPV continues to be a significant public health problem and represents an area of health disparity for ethnic minorities. PMID- 15361584 TI - Arabidopsis RIN4 negatively regulates disease resistance mediated by RPS2 and RPM1 downstream or independent of the NDR1 signal modulator and is not required for the virulence functions of bacterial type III effectors AvrRpt2 or AvrRpm1. AB - Bacterial pathogens deliver type III effector proteins into the plant cell during infection. On susceptible (r) hosts, type III effectors can contribute to virulence. Some trigger the action of specific disease resistance (R) gene products. The activation of R proteins can occur indirectly via modification of a host target. Thus, at least some type III effectors are recognized at site(s) where they may act as virulence factors. These data indicate that a type III effector's host target might be required for both initiation of R function in resistant plants and pathogen virulence in susceptible plants. In Arabidopsis thaliana, RPM1-interacting protein 4 (RIN4) associates with both the Resistance to Pseudomonas syringae pv maculicola 1 (RPM1) and Resistance to P. syringae 2 (RPS2) disease resistance proteins. RIN4 is posttranslationally modified after delivery of the P. syringae type III effectors AvrRpm1, AvrB, or AvrRpt2 to plant cells. Thus, RIN4 may be a target for virulence functions of these type III effectors. We demonstrate that RIN4 is not the only host target for AvrRpm1 and AvrRpt2 in susceptible plants because its elimination does not diminish their virulence functions. In fact, RIN4 negatively regulates AvrRpt2 virulence function. RIN4 also negatively regulates inappropriate activation of both RPM1 and RPS2. Inappropriate activation of RPS2 is nonspecific disease resistance 1 (NDR1) independent, in contrast with the established requirement for NDR1 during AvrRpt2-dependent RPS2 activation. Thus, RIN4 acts either cooperatively, downstream, or independently of NDR1 to negatively regulate RPS2 in the absence of pathogen. We propose that many P. syringae type III effectors have more than one target in the host cell. We suggest that a limited set of these targets, perhaps only one, are associated with R proteins. Thus, whereas any pathogen virulence factor may have multiple targets, the perturbation of only one is necessary and sufficient for R activation. PMID- 15361586 TI - The impact of culture and minority status on women's experience of domestic violence. AB - The influence of culture and ethnic background on women's experience of domestic violence has been explored in research only recently. Here the authors review research about the impact of culture and minority status in the United States on women's experience of domestic violence, considering family structure,immigration, acculturation, oppression, and community response. The authors encourage researchers and service providers to acknowledge the effects on women of sociopolitical dynamics, including racism, and to identify specific aspects of culture that are relevant to intimate partner abuse. PMID- 15361587 TI - The effects of child maltreatment on the hypothalamic-pituitary-adrenal axis. AB - Abnormal functioning of the hypothalamic-pituitary-adrenal (HPA) axis, a critical mammalian stress response system, has been associated with emotional responses such as anxiety and depression, as well as with behavioral and cognitive processes such as aggression, learning and memory deficits, and failure of response inhibition. This review examines the evidence for HPA axis dysregulation related to childhood maltreatment. It is concluded that child maltreatment may lead to disruptions in HPA axis functioning, and that factors such as age of maltreatment, parental responsiveness, subsequent exposure to stressors, type of maltreatment, and type of psychopathology or behavioral disturbance displayed may influence the degree and patterning of HPA disturbance. PMID- 15361588 TI - Intimate partner violence and sexual assault in Native American communities. AB - Previous studies indicate that Native American women experience the highest rate of violence of any ethnic or racial group in the United States. This article addresses the prevalence of intimate partner violence and sexual assault among Native Americans. We present significant substantive and methodological issues that inform research on violence in the lives of Native Americans, as well as existing interventions. Interventions discussed in this article fall within three major categories including those that are community based, those grounded in the public health and health care systems, and those grounded in federal and national organizations. We provide some examples of interventions from each of these three levels of direct service, including a brief discussion of barriers to service accessibility. We conclude with substantive and methodological recommendations for research and practice. PMID- 15361589 TI - International science meetings. PMID- 15361590 TI - Stem cell politics. California debates whether to become stem cell heavyweight. PMID- 15361591 TI - AIDS vaccines. HIV dodges one-two punch. PMID- 15361592 TI - Japan budget. Science ministry puts in for big increases. PMID- 15361593 TI - Climate change. Changes in planktonic food web hint at major disruptions in Atlantic. PMID- 15361594 TI - Scientific publishing. NIH proposes 6-month public access to papers. PMID- 15361595 TI - Nuclear proliferation. South Korea admits to laser enrichment program. PMID- 15361596 TI - European Union. The commissioner who listened. PMID- 15361597 TI - Science policy. The man behind the memos. PMID- 15361598 TI - Endangered Species Act. Can California ranchers save the tiger salamander? PMID- 15361599 TI - Brian Fagan profile. Archaeologist leaves an imprint on his field--without research. PMID- 15361600 TI - Ocean ecology. Dead zone fix not a dead issue. PMID- 15361601 TI - American Chemical Society meeting. Finding reactions in a haystack: try 'em all, see what works. PMID- 15361602 TI - American Chemical Society meeting. Enzyme deactivates heart-friendly HDL. PMID- 15361603 TI - American Chemical Society meeting. Breaking a barrier to new brain images. PMID- 15361604 TI - American Chemical Society meeting. Snapshots from the meeting. PMID- 15361605 TI - Extinction rates and butterflies. PMID- 15361606 TI - Noguchi's contributions to science. PMID- 15361607 TI - Networks by design: a revolution in ecology. PMID- 15361608 TI - Comment on "Managing soil carbon" (I). PMID- 15361609 TI - Comment on "Managing soil carbon" (II). PMID- 15361611 TI - Biotechnology. Japanese controversies over transgenic crop regulation. PMID- 15361612 TI - Structural biology. The atomic architecture of a gas channel. PMID- 15361613 TI - Physics. Visualizing the dynamics of the onset of turbulence. PMID- 15361614 TI - Chemistry. Multidimensional snapshots of chemical dynamics. PMID- 15361615 TI - Microbiology. Pathogenic bacteria prefer heme. PMID- 15361616 TI - Microbiology. Noninherited resistance to antibiotics. PMID- 15361617 TI - Asymmetric cochlear processing mimics hemispheric specialization. AB - Otoacoustic emissions or OAEs (reflections of cochlear energy produced during the processing of sound) were measured in response to two types of stimuli, rapid clicks and sustained tones, in each ear of neonates. OAEs were larger to tones when elicited in the left ear and to clicks when elicited in the right. This finding is similar to those of enhanced processing of tones in right auditory cortical areas and of rapid stimuli on the left, given strong crossed connections from ear to brain. These findings indicate that processing at the level of the ear may facilitate lateralization of auditory function in the brain. PMID- 15361619 TI - Experimental observation of nonlinear traveling waves in turbulent pipe flow. AB - Transition to turbulence in pipe flow is one of the most fundamental and longest standing problems in fluid dynamics. Stability theory suggests that the flow remains laminar for all flow rates, but in practice pipe flow becomes turbulent even at moderate speeds. This transition drastically affects the transport efficiency of mass, momentum, and heat. On the basis of the recent discovery of unstable traveling waves in computational studies of the Navier-Stokes equations and ideas from dynamical systems theory, a model for the transition process has been suggested. We report experimental observation of these traveling waves in pipe flow, confirming the proposed transition scenario and suggesting that the dynamics associated with these unstable states may indeed capture the nature of fluid turbulence. PMID- 15361618 TI - Mechanism of ammonia transport by Amt/MEP/Rh: structure of AmtB at 1.35 A. AB - The first structure of an ammonia channel from the Amt/MEP/Rh protein superfamily, determined to 1.35 angstrom resolution, shows it to be a channel that spans the membrane 11 times. Two structurally similar halves span the membrane with opposite polarity. Structures with and without ammonia or methyl ammonia show a vestibule that recruits NH4+/NH3, a binding site for NH4+, and a 20 angstrom-long hydrophobic channel that lowers the NH4+ pKa to below 6 and conducts NH3. Favorable interactions for NH3 are seen within the channel and use conserved histidines. Reconstitution of AmtB into vesicles shows that AmtB conducts uncharged NH3. PMID- 15361620 TI - Mesophase structure-mechanical and ionic transport correlations in extended amphiphilic dendrons. AB - We have studied the self-assembly of amphiphilic dendrons extended with linear polyethylene oxide (PEO) chains and their ion complexes. Keeping the dendron core and linear PEO chain compatible allows for the combination of dendritic core shell and conventional blockcopolymer characteristics for complex mesophase behavior. An unexpected sequence of crystalline lamellar, cubic micellar (Pm3n), hexagonal columnar, continuous cubic (Ia3d), and lamellar mesophases is observed. Multiple phase behavior within single compounds allows for the study of charge transport and mechanical property correlations as a function of structure. The results suggest an advanced molecular design concept for the next generation of nanostructured materials in applications involving charge transport. PMID- 15361621 TI - Hydrophobic collapse in multidomain protein folding. AB - We performed molecular dynamics simulations of the collapse of a two-domain protein, the BphC enzyme, into a globular structure to examine how water molecules mediate hydrophobic collapse of proteins. In the interdomain region, liquid water persists with a density 10 to 15% lower than in the bulk, even at small domain separations. Water depletion and hydrophobic collapse occur on a nanosecond time scale, which is two orders of magnitude slower than that found in the collapse of idealized paraffin-like plates. When the electrostatic protein water forces are turned off, a dewetting transition occurs in the interdomain region and the collapse speeds up by more than an order of magnitude. When attractive van der Waals forces are turned off as well, the dewetting in the interdomain region is more profound, and the collapse is even faster. PMID- 15361622 TI - Climate impact on plankton ecosystems in the Northeast Atlantic. AB - It is now widely accepted that global warming is occurring, yet its effects on the world's largest ecosystem, the marine pelagic realm, are largely unknown. We show that sea surface warming in the Northeast Atlantic is accompanied by increasing phytoplankton abundance in cooler regions and decreasing phytoplankton abundance in warmer regions. This impact propagates up the food web (bottom-up control) through copepod herbivores to zooplankton carnivores because of tight trophic coupling. Future warming is therefore likely to alter the spatial distribution of primary and secondary pelagic production, affecting ecosystem services and placing additional stress on already-depleted fish and mammal populations. PMID- 15361623 TI - Methanobactin, a copper-acquisition compound from methane-oxidizing bacteria. AB - Siderophores are extracellular iron-binding compounds that mediate iron transport into many cells. We present evidence of analogous molecules for copper transport from methane-oxidizing bacteria, represented here by a small fluorescent chromopeptide (C45N12O14H62Cu, 1216 daltons) produced by Methylosinus trichosporium OB3b. The crystal structure of this compound, methanobactin, was resolved to 1.15 angstroms. It is composed of a tetrapeptide, a tripeptide, and several unusual moieties, including two 4-thionyl-5-hydroxy-imidazole chromophores that coordinate the copper, a pyrrolidine that confers a bend in the overall chain, and an amino-terminal isopropylester group. The copper coordination environment includes a dual nitrogen- and sulfur-donating system derived from the thionyl imidazolate moieties. Structural elucidation of this molecule has broad implications in terms of organo-copper chemistry, biological methane oxidation, and global carbon cycling. PMID- 15361624 TI - Activation of endogenous Cdc42 visualized in living cells. AB - Signaling proteins are tightly regulated spatially and temporally to perform multiple functions. For Cdc42 and other guanosine triphosphatases, the subcellular location of activation is a critical determinant of cell behavior. However, current approaches are limited in their ability to examine the dynamics of Cdc42 activity in living cells. We report the development of a biosensor capable of visualizing the changing activation of endogenous, unlabeled Cdc42 in living cells. With the use of a dye that reports protein interactions, the biosensor revealed localized activation in the trans-Golgi apparatus, microtubule dependent Cdc42 activation at the cell periphery, and activation kinetics precisely coordinated with cell extension and retraction. PMID- 15361625 TI - Advanced cardiac morphogenesis does not require heart tube fusion. AB - The bilateral cardiac mesoderm migrates from the lateral region of the embryo to the ventral midline, where it fuses to form the primitive heart tube. It is generally accepted that migration and fusion are essential for subsequent stages of cardiac morphogenesis. We present evidence that, in Foxp4 mutant embryonic mice, each bilateral heart-forming region is capable of developing into a highly differentiated four-chambered mammalian heart in the absence of midline fusion. These data demonstrate that left-right chamber specification, cardiac looping, septation, cardiac myocyte differentiation, and endocardial cushion formation are preprogrammed in the precardiac mesoderm and do not require midline positional identity or heart tube fusion. PMID- 15361626 TI - Iron-source preference of Staphylococcus aureus infections. AB - Although bacteria use different iron compounds in vitGro, the possibility that microbes distinguish between these iron sources during infection has hitherto not been examined. We applied stable isotope labeling to detect source-specific iron by mass spectrometry and show that Staphylococcus aureus preferentially imports heme iron over transferrin iron. By combining this approach with computational genome analysis, we identified hts (heme transport system), a gene cluster that promotes preferred heme iron import by S. aureus. Heme iron scavenging by means of hts is required for staphylococcal pathogenesis in animal hosts, indicating that heme iron is the preferred iron source during the initiation of infection. PMID- 15361627 TI - Species coextinctions and the biodiversity crisis. AB - To assess the coextinction of species (the loss of a species upon the loss of another), we present a probabilistic model, scaled with empirical data. The model examines the relationship between coextinction levels (proportion of species extinct) of affiliates and their hosts across a wide range of coevolved interspecific systems: pollinating Ficus wasps and Ficus, parasites and their hosts, butterflies and their larval host plants, and ant butterflies and their host ants. Applying a nomographic method based on mean host specificity (number of host species per affiliate species), we estimate that 6300 affiliate species are "coendangered" with host species currently listed as endangered. Current extinction estimates need to be recalibrated by taking species coextinctions into account. PMID- 15361629 TI - Silicone sheeting as an alternative to elastic bandages in dressing lower extremity amputations. PMID- 15361630 TI - Empowering the ileostomy patient with a mechanical bowel obstruction. PMID- 15361631 TI - Continent diversions: the new gold standards of ileoanal reservoir and neobladder. AB - In recent decades, surgical treatment of familial adenomatous polyposis, chronic ulcerative colitis, and muscle-invasive bladder cancer has undergone a revolution. Specifically, ileoanal reservoir and neobladder have become the new "gold standard" of definitive surgical therapy for these disorders. This article discusses issues in surgical construction, indications, contraindications, perioperative care concepts, and nursing and health professional implications related to these two procedures. These interventions include screening candidates for ileoanal reservoir or neobladder to rule out Crohn's disease or metastatic cancer and educating candidates for continent diversions about the proposed procedure(s) and associated events, potential complications, postoperative exercise, sexual health and function issues, and the benefits of support group participation so they can gain a realistic understanding of ultimate functional outcomes. Questions for future research are addressed. PMID- 15361632 TI - Challenges of ostomy care and obesity. AB - In the United States, 13 to 16 million people are morbidly obese to the extent that medical intervention, simply based on obesity-related comorbidities, is needed. The challenge of ostomy care and the very obese patient lies in the skill and planning required for successful physical, emotional, and spiritual recovery. It is commonly believed that from the onset, the obese patient having surgery is at a significant disadvantage. Care of the obese patient requiring ostomy surgery includes considerable challenges - from preoperative preparation, including finding an optimal location for stoma placement, to the challenge of preventing complications during the intraoperative and postoperative phases of care. Concerns regarding pain management, immobility, skin injury, respiratory issues, embolic threats, and caregiver injury increase when treating the obese patient and must be addressed specifically. A case study approach is used as a framework to discuss the ostomy experience. PMID- 15361633 TI - Assessment and management of stomal complications: a framework for clinical decision making. AB - Assessment and management of stoma complications are often the responsibility of nurses across the continuum of care. These complications can occur at different times based on their etiology - immediately postoperatively or even several years after surgery - and often require modifications in a person's daily stoma management. This article presents a conceptual framework to help categorize types of stoma complications based on either etiology or location and offers management options to facilitate quality care. The five major categories of complications include Poor Siting, Stoma Proper, Peri-Intestinal Area, Mucocutaneous Junction, and Iatrogenic. Most of these suggested approaches to care are the recommendations of certified ostomy nurses based on their educational training, expert opinion, and successful experiences. Although these recommendations have often solved the specific problems and greatly improved the quality of life for the person with stomal complications, much research is still needed to confirm and/or improve these nursing approaches. PMID- 15361634 TI - Peristomal skin complications: prevention and management. AB - Peristomal skin complications are the most common reason ostomy patients visit an outpatient wound, ostomy, and continence nursing service. Prevention and management of peristomal skin complications are critical components of ostomy care. Identifying risk factors for the occurrence of peristomal skin complications according to types of injury and clinical features can help optimize assessment and management approaches. Treatment can further be addressed based on etiology - chemical injury (irritant contact dermatitis, pseudoverrucous lesions, and encrustations); mechanical injury (pressure/shear, stripping, mucocutaneous separation, mucosal transplantation); infection (Candidiasis, folliculitis); immunologic disorders (allergic contact dermatitis); and disease related lesions (varices, pyoderma gangrenosum, malignancy). The importance of prevention and the impact of having access to knowledgeable care providers cannot be over-emphasized. PMID- 15361636 TI - An unusual presentation of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) metastasis to the lymph nodes has been reported in about one-third of autopsies, mostly to the regional chains. Involvement of cervical lymph nodes is extremely rare. We encountered an unusual case of HCC in a patient with chronic liver disease presenting with right-sided anterior cervical lymph node metastasis skipping mediastinal and supraclavicular chains. One should be aware of that unusual site of presentation and metastasis of HCC so as not to miss the diagnosis. PMID- 15361637 TI - Acinar cell carcinoma of the pancreas. AB - Acinar cell carcinoma of the pancreas is a rare tumor for which the best chemotherapy regimen has not been clearly established. Here, we report on a female patient with an unusually long survival and a remarkable response to weekly paclitaxel. To our knowledge, this is the first time that paclitaxel has been associated with an objective response in this disease. The patient recurred after initial resection and failed multiple prior chemotherapeutic regimens. She received weekly paclitaxel at 80 mg/m2 and responded after eight doses, maintaining controlled disease for a total of 4 mo. We present her case in detail and review the available literature regarding this rare type of tumor. PMID- 15361638 TI - Colon cancer with isolated metastasis to the kidney at the time of initial diagnosis. AB - Blood-borne metastases to the kidneys from solid tumors have received little attention in the medical literature because they usually occur in a setting of advanced systemic disease, and renal involvement is a relatively minor cause of symptoms. Although the frequency of metastases to the kidney in cancer patients is 7-13% in large autopsy series, incidental discovery of a renal metastasis as the first manifestation of a primary tumor is a very rare event. The most common primary malignancy to involve the kidney is bronchogenic carcinoma,followed by breast and gastrointestinal cancers. In this article, we report a patient with left colon cancer and isolated metastasis to the right kidney at the time of initial diagnosis. Left hemicolectomy and right nephrectomy were performed. Adjuvant systemic chemotherapy consisting of 5-fluorouracil (5-FU) and folinic acid (FA) was given. 5-FU and FA were stopped after four cycles because metastases to the lung and liver occurred about 3 mo after the surgery during adjuvant chemotherapy. Capecitabine was started. The patient died 9 mo after the discovery of the isolated renal metastasis. Nephrectomy is more for diagnostic clarification in the setting of synchronous primary because it has no effect on survival and its effect on quality of life is minimal; as seen in our case, the other organ metastases rapidly occur and the survival is limited. Nephrectomy may also compromise the choice of chemotherapy agents that require renal clearance, thus a careful evaluation of renal functions is necessary if a nephrectomy is performed. In the matter of a decreased renal clearance,the doses of these drugs should be decreased or the choice should be reevaluated. PMID- 15361639 TI - Phase II clinical trial of 5-fluorouracil, trimetrexate, and leucovorin (NFL) in patients with advanced pancreatic cancer. AB - BACKGROUND: Advanced pancreatic cancer has limited treatment options. 5 fluorouracil (5-FU) is frequently used in the treatment of pancreatic cancer. Preclinical studies suggest synergism between trimetrexate (TMTX),5-FU, and leucovorin (NFL). AIM: We conducted a phase II trial to evaluate the activity and safety of NFL in pancreatic cancer. METHOD: Eligible patients (n = 21) with untreated advanced pancreatic cancer were treated with 110 mg/m2 intravenous (IV) THTX on day 1 and 200 mg/m2 IV leucovorin prior to 500 mg/m2 IV 5-FU on day 2. Oral leucovorin (15 mg every 6 h for seven doses) started intravenous 24 h later. RESULTS: Treatment was administered for 6 wk followed by a 2-wk rest period. Response was evaluated every 8 wk. All patients were evaluable for response and toxicity. Most patients (80%) had distant metastases. Forty-five cycles of chemotherapy were administered. The most common serious toxicities were Grade 3 diarrhea (23.8%) and nausea and vomiting (14.2%). The response rate was 4.1% (95% CI, 0-23%), median survival was 6.8 mo, and 1-yr survival was 19%. CONCLUSION: Treatment with NFL is well-tolerated in patients with advanced pancreatic cancer. The median survival and 1-yr survival in these patients with poor prognosis compares favorably with other treatment options. PMID- 15361640 TI - Evaluation of epstein-barr virus DNA load in gastric mucosa with chronic atrophic gastritis using a real-time quantitative PCR assay. AB - PURPOSE: It is hypothesized that Epstein-Barr virus (EBV) has already infected the noncarcinomatous gastric mucosa before carcinogenesis of EBV-associated gastric carcinoma. However, the frequency and distribution of EBV infection in the gastric mucosa of chronic atrophic gastritis (CAG) are still unclear. To clarify these points, we evaluated the EBV DNA load in gastric mucosa with CAG. METHODS: We tested samples from 35 CAG cases. Paired biopsy specimens from five sites of the stomach were obtained according to the Updated Sydney System. One of each pair of specimens was subjected to areal-time quantitative polymerase chain reaction (Q-PCR) assay to detect EBV. Q-PCR was performed using the LightCycler System (Roche, Mannheim, Germany). The other was subjected to hematoxylin and eosin (H&E) and Giemsa staining. The histological degree of CAG was graded according to the Updated Sydney System. To evaluate the surface distribution of gastric mucosal atrophic changes of CAG, we modified the endoscopic classification of Kimura and Takemoto. RESULT: EBV DNA was detected in 65.7% (23 of 35 cases) of the gastric biopsy specimens of the cases examined. EBV DNA was detected most frequently (92.3%; 12 of 13 cases) in the cases with endoscopically moderate CAG (p < 0.01). There was a significant association between EBV detection and the presence of inflammatory cell infiltration and atrophy in the stomach with endoscopically moderate CAG. CONCLUSION: EBV mainly infects the gastric mucosa of patients with moderate CAG. PMID- 15361641 TI - Basaloid squamous cell cancer arising in Barrett's esophagus. AB - Basaloid squamous cell carcinoma (BSCC) is a rare form of cancer that arises primarily in the upper aerodigestive tract. Esophageal BSCC is extremely rare, accounting for less than 2% of primary esophageal malignancies. It is histopathologically distinct from squamous cell carcinoma and has an aggressive biological behavior with poor survival outcomes. There is no known association of Barrett's esophagus with esophageal BSCC. Here, we report what we believe is the first such case of esophageal BSCC occurring in the setting of Barrett's esophagus. PMID- 15361642 TI - Biliopancreatic fistula caused by an intraductal papillary-mucinous tumor of the pancreas confirmed by biochemical analysis of mucin. AB - Intraductal papillary-mucinous tumor of the pancreas is occasionally accompanied by biliopancreatic fistula. However, it is difficult to show the inflow of mucin produced by the tumor into the common bile duct. To confirm the biliopancreatic fistula, the mucin-rich fraction was purified from the bile and stained with antimucin antibodies. Western blot analysis showed characteristic smear staining patterns for mucin molecules with three types of antimucin antibodies. Immunohistochemical analysis with the antibody showed significant signals of the cancer cells and the luminal content of the dilated pancreatic duct. These results showed that the bile contained an abundance of mucin, which was produced by the primary pancreatic tumor. In cases with intraductal papillary-mucinous tumor of the pancreas, biochemical analysis of mucin molecules in the bile can be of clinical use in consideration of pathological process of tumor progression. PMID- 15361643 TI - Cancer and leukemia group B (CALGB) 89805: phase II chemoradiation trial using gemcitabine in patients with locoregional adenocarcinoma of the pancreas. AB - PURPOSE: Determine the safety and efficacy of twice weekly gemcitabine and concurrent radiation to the upper abdomen followed by weekly gemcitabine in patients with surgically staged, locally advanced pancreatic cancer. METHODS: Patients with surgically staged, locally advanced, nonmetastatic adenocarcinoma of the pancreas were treated with intravenous gemcitabine administered twice weekly (40 mg/m2/d) for 5 wk concurrent with upper abdominal radiation (50.4 Gy in 180 cGy daily fractions over 5.5 wk). At the completion of the chemoradiation, patients without disease progression were given gemcitabine (1000 mg/m2) weekly for five cycles. Each cycle consisted of 3 wk of treatment followed by 1 wk without treatment. Disease progression and response were assessed at 6- to 8-wk intervals. RESULTS: From February through December 1999, 43 patients were entered into this phase II trial, 39 of whom were evaluable for treatment response. The median age was 59 yr (range: 39-84 yr); there were 18 males (47%) in the study. Grade III and IV hematologic toxicity occurred in 48 and 21% of patients, respectively, and was primarily leukocytopenia and neutropenia. Grade III and IV gastrointestinal toxicities occurred in 31 and 10% of patients, respectively. There was one death attributed to sepsis. The concurrent gemcitabine and radiation portion of the study was completed without treatment interruptions in 56% of patients. The overall median survival was 8.2 mo and the median survival in the 44% of patients demonstrating a sustained CA-19-9 response was 13.5 mo. Only six patients experienced local regional progression as their first site of failure. Two patients (5%) were still alive at 35 and 41 mo posttreatment. CONCLUSIONS: These results confirm the feasibility of twice weekly gemcitabine and radiation for the treatment of pancreatic cancer. Although this treatment strategy produced good local regional control, this did not result in a survival advantage. Stratifying patients by performance status and CA-19-9 response in future trials may be of value. PMID- 15361644 TI - Carcinoma of the cystic duct remnant with direct colonic invasion. AB - Carcinoma of the cystic duct remnant is extremely rare, and only five cases have been reported previously in the English literature. We present another case of carcinoma of the cystic duct remnant and review the literature. A 62-yr-old woman was diagnosed as having hilar cholangiocarcinoma and transverse colon cancer. However, at laparotomy, the center of the tumor mass was in the cystic duct remnant with contiguous colonic invasion. The pathology of the surgical specimen confirmed carcinoma of the cystic duct remnant. The diagnosis of carcinoma of the cystic duct remnant was made using Ozden's new criteria. PMID- 15361645 TI - Neoadjuvant chemoradiotherapy for adenocarcinoma of the pancreas: analysis of histopathology and outcome. AB - To examine the histopathologic effect of neoadjuvant therapy and its impact on survival in patients with carcinoma of the pancreas, we retrospectively reviewed the records of 116 patients who underwent resections for pancreatic cancer from 1987 to 2000. Median follow-up of surviving patients was 19 mo(range 4-150 mo). Preoperative chemotherapy was administered in 61 patients (53%) and consisted of 5-fluorouracil/mitomycin C in 35 patients and gemcitabine in 26 patients, given concurrently with external beam radiation (5040 cGy). All resections were performed with curative intent (98 Whipples, 11 total, 6 distal, and 1 central pancreatectomy). Histopathologic examination included an estimation of the amount of fibrosis present in the tumor specimen (expressed as the percentage of fibrosis identified relative to the amount of neoplastic cells present). The mean fibrosis level for the series was 56% (range 5% to 100%). The administration of neoadjuvant therapy resulted in greater fibrosis (73%) than no preoperative treatment (38%) (p = 0.0001). Higher mean fibrosis levels were observed in patients with negative lymph nodes (p = 0.0006) and negative margins (p = 0.05). Factors associated with improved survival(log rank test) included: negative margins (p = 0.001), negative lymph nodes (p = 0.03), and use of neoadjuvant therapy (p = 0.03). Median survival in the neoadjuvant group was 23 mo vs 16 mo without preoperative therapy (p = 0.03). In conclusion, the use of neoadjuvant therapy resulted in a greater degree of fibrosis in the specimen. Patients with negative margins and negative lymph nodes had a greater amount of fibrosis present, and these were significant predictors of improved outcome. Although retrospective,this series suggests an improvement in survival in patients treated with neoadjuvant therapy. PMID- 15361646 TI - Local recurrence of rectal cancer after total mesorectal excision without preoperative radiotherapy. AB - BACKGROUND: At this moment, it is still debatable whether all patients with mobile rectal cancer who undergo surgical removal of the tumor should be treated with preoperative radiotherapy, since it is likely that only certain patients will benefit from this strategy. In this study, patients with mobile rectal cancer were immediately operated upon and only those with positive nodes or with incomplete resection received adjuvant radiotherapy. AIMS OF THE STUDY: To investigate the local recurrence rate after the use of a selective policy of adjuvant radiotherapy and to determine risk factors for local recurrence. METHODS: In a 5-yr-period, 178 patients with rectal cancer were referred to our institute. A total of 131 patients with mobile rectal cancer were treated with curative intent, which implied a microscopically radical resection and no signs of distant metastasis at operation. A retrospective analysis was undertaken to investigate the incidence of local recurrence in this curative group and to determine risk factors for local recurrence. RESULTS: The postoperative mortality in the curative group was 5.3%. Local recurrences were observed in 6 patients (4.6%) after a median period of 25 mo (range 11-37); two of them also had distant metastases detected at the same time. The highest local recurrence rates were seen in men (5.3%), in distal rectal cancers (6.9%), and in the node-positive group (8.7%). CONCLUSION: A low local recurrence rate can be achieved after total mesorectal excision (TME) without preoperative radiotherapy. Our results suggest using preoperative radiotherapy only for those patients who are at a higher risk for local recurrence. Staging techniques for selection of these patients are at this moments till inappropriate. PMID- 15361647 TI - Sub-acute sensory neuronopathy as a preceding sign of recurrence in colon carcinoma. AB - Subacute sensory neuronopathy is a paraneoplastic syndrome, which occurs mostly in lung, breast, ovarian malignancies and lymphoma. A 75-yr-old woman who was at the twentieth month of her postoperative follow-up owing to colon adenocarcinoma admitted with subacute sensory neuronopathy. Six months later from the first, neuropathic symptoms liver metastases developed. To the best of our literature review subacute sensory neuronopathy as a preceding sign of recurrence in colon adenocarcinoma has not previously been reported. We conclude that, in the case of subacute sensory neuronopathy without an obvious underlying etiological factor, an occult malignity should always be researched in clinical practice. PMID- 15361648 TI - Carcinoma of the esophagus with mixed basaloid squamous and glandular differentiation: a distinct histological presentation. AB - Esophageal cancer is the third most common gastrointestinal cancer and ranks among the ten commonest cancers worldwide. Histologically, approx 60% of esophageal cancers are adenocarcinomas and 40% are squamous cell carcinomas (SCC). Other rare cancers of the esophagus include small-cell carcinomas, squamous cell carcinomas with sarcomatous features, adenoid cystic carcinomas, and mucoepidermoid carcinomas. Basaloid squamous cell carcinoma (BSCC) or basaloid squamous carcinoma (BSC) is a distinct clinicopathological entity, seen more frequently in elderly males. Stage at presentation is often advanced and regional adenopathy or distant metastases are not uncommon at presentation. We describe an unusual case report of esophageal BSCC with glandular differentiation. The clinical significance of glandular differentiation in this rare type of tumor is not known. PMID- 15361649 TI - Fish oil supplementation in the treatment of cachexia in pancreatic cancer patients. AB - Patients with pancreatic cancer often experience a loss of weight and appetite, known as the anorexia-cachexia syndrome, which is associated with decreased quality of life and reduced survival. Research into the biological mechanisms of cachexia has demonstrated that an array of inflammatory mediators and tumor derived factors cause appetite suppression, skeletal muscle proteolysis, and lipolysis,producing an overall hypercatabolic state that contributes to loss of fat and lean body mass. Omega-3 polyunsaturated fatty acids (n-3 PUFAs) have been shown to modulate levels of proinflammatory cytokines, hepatic acute phase proteins, eicosanoids, and tumor-derived factors in animal models of cancer and may reverse some aspects of the process of cachexia. Results of clinical trials of n-3 PUFAs in the form of fish oils have been mixed, but should encourage further investigation into dietary fish oil supplementation, including the most effective route of administration and the proper dosage to promote optimal weight maintenance and to limit side effects. Concerns about standardization and quality control should also be considered. With the current available evidence, a recommendation for the use of omega 3 polyunsaturated fatty acids in pancreatic cancer cachexia is premature. PMID- 15361650 TI - Gallbladder cystadenoma and common bile duct obstruction. AB - Cystadenomas are usually found in the extra-hepatic bile ducts in conjunction with multilocular cysts in the liver. Cystadenoma of the gallbladder itself is a rare finding, cited only once in the literature as the cause of extrinsic obstruction of the common bile duct (5). In this report, we describe the endoscopic retrograde cholangiopancreaticographic (ERCP) detection of intrinsic obstruction of the cystic duct and common bile duct by such a tumor in a 47-yr old woman. PMID- 15361651 TI - Culture of primary adherent cells and a continuously growing nonadherent cell line. AB - Cell culture is an invaluable tool for investigators in numerous fields. It facilitates analysis of biological properties and processes that are not readily accessible at the level of the intact organism. Successful maintenance of cells in culture, whether primary or immortalized, requires knowledge and practice of a few essential techniques. The purpose of this chapter is to explain the basic principles of cell culture using the maintenance of a nonadherent cell line, the P815 mouse mastocytoma cell line, and the isolation and culture of adherent primary mouse embryonic fibroblasts (MEFs) as examples. Procedures for thawing, culture, determination of cell numbers and viability, and cryopreservation are described. PMID- 15361652 TI - Detection of mycoplasma contaminations. AB - Mycoplasma contamination of cell lines is one of the major problems in cell culture technology. The specific, sensitive, and reliable detection of mycoplasma contamination is an important part of mycoplasma control and should be an established method in every cell culture laboratory. New cell lines as well as cell lines in continuous culture must be tested in regular intervals. The polymerase chain reaction (PCR) methodology offers a fast and sensitive technique to monitor all cultures in a laboratory. The technique can also be used to determine the contaminating mycoplasma species. The described assay can be performed within 3 h, including sample preparation, DNA extraction, performing the PCR reaction, and analysis of the PCR products. Special precautions necessary to avoid false-negative results resulting from inhibitors of the Taq polymerase present in the crude samples and the interpretation of the results are also described. PMID- 15361653 TI - Eradication of mycoplasma contaminations. AB - Mycoplasma contaminations have a multitude of effects on cultured cell lines that can potentially influence the results of experiments and pollute bioactive substances used in human medicine. The elimination of mycoplasma contaminations in cell cultures has become a practical alternative to discarding and re establishing important or irreplaceable cell lines. Different quinolones, tetracyclins, and macrolides shown to have strong antimycoplasma properties are employed for the decontamination. We provide detailed descriptions to assure eradication of mycoplasma, to prevent formation of resistant mycoplasma strains, and to cure heavily contaminated and damaged cells. To date, no consistent and permanent alterations that affect the eukaryotic cells during or after the treatment have been detected. PMID- 15361654 TI - Authentication of scientific human cell lines: easy-to-use DNA fingerprinting. AB - Human cell lines are an important resource for research and most often used in reverse genetic approaches or as in vitro model systems of human diseases. In this regard, it is crucial that the cells faithfully correspond to the purported objects of study. A number of recent publications have shown an unacceptable level of cell lines to be false, in part as a result of the nonavailability of a simple and easy DNA profiling technique. We have validated different single- and multiple-locus variable numbers of tandem repeats (VNTRs) enabling the establishment of a noncommercial, but good laboratory practice, method for authentication of cell lines by DNA fingerprinting. Polymerase chain reaction amplification fragment length polymorphism (AmpFLP) of six prominent and highly polymorphic minisatellite VNTR loci, requiring only a thermal cycler and an electrophoretic system, was proven as the most reliable tool. Furthermore, the generated banding pattern and the determination of gender allows for verifying the authenticity of a given human cell line by simple agarose gel electrophoresis. The combination of rapidly generated DNA profiles based on single-locus VNTR loci and information on banding patterns of cell lines of interest by official cell banks (detailed information at the website www.dsmz.de) constitute a low-cost but highly reliable and robust method, enabling every researcher using human cell lines to easily verify cell line identity. PMID- 15361655 TI - Cytogenetic analysis of cell lines. AB - Cytogenetic analysis forms an essential part of characterizing and identifying cell lines, in particular those established from tumors. In addition, karyotypic analysis can be used to distinguish individual subclones and to monitor stability. This chapter describes basic cytogenetic procedures suited to cells in continuous culture. The provision of unlimited material by cell lines encourages an heuristic approach to harvesting and hypotonic treatments to yield metaphase chromosome slide preparations of improved quality suitable for subsequent banding and fluorescence in situ hybridization (FISH) analysis. The experience of the writers with more than 500 different cell lines has shown that no single hypotonic harvesting protocol is adequate to consistently deliver satisfactory chromosome preparations. Thus, evidence-based protocols are described for hypotonic harvesting, rapid G-banding, and FISH analysis of cell cultures to allow troubleshooting and fine-tuning to suit the requirements of individual cell lines. PMID- 15361656 TI - Human and mouse hematopoietic colony-forming cell assays. AB - Hematopoietic stem cells present in small numbers in certain fetal organs during development and in adult bone marrow produce a heterogeneous pool of progenitors that can be detected in vitro using colony-forming cell (CFC) assays. Hematopoietic progenitor cells, when cultured in a semisolid methylcellulose based medium that is supplemented with suitable growth factors, proliferate and differentiate to produce clonal clusters (colonies) of maturing cells. The CFCs are then classified and enumerated in situ by light microscopy. Protocols for the detection and enumeration of myeloid multipotential progenitors and committed progenitors of the erythroid, monocyte, and granulocyte lineages in samples from human peripheral blood, bone marrow, and cord blood as well as mouse fetal liver and bone marrow are described. PMID- 15361658 TI - Isolation and culture of human macrophages. AB - Methods to isolate and culture human monocyte-derived macrophages and alveolar macrophages are described. Monocytes are obtained from buffy-coat preparations by Ficoll density gradient centrifugation, followed by adhesion-mediated purification on tissue culture or gelatin-coated plastic. The monocytes differentiate into macrophages in vitro by culturing in medium containing autologous human fibrin-depleted plasma. Alveolar macrophages can be purified from bronchoalveolar fluid samples by adhesion to tissue culture plastic. If resected lung tissue is available, alveolar macrophages can be obtained by mechanically disrupting the lung parenchyma, followed by adhesion-mediated purification. PMID- 15361657 TI - Isolation and culture of murine macrophages. AB - The two most convenient sources of primary murine macrophages are the bone marrow and the peritoneal cavity. Resident peritoneal macrophages can readily be harvested from mice and purified by adherence to tissue culture plastic. The injection of Bio-Gel polyacrylamide beads or thioglycollate broth into the peritoneal cavity produces an inflammatory response allowing the purification of large numbers of elicited macrophages. The production of an activated macrophage population can be achieved by using Bacillus-Calmette-Guerin as the inflammatory stimulus. Resident bone marrow macrophages can be isolated following enzymatic separation of cells from bone marrow plugs and enrichment on 30% fetal calf serum containing medium or Ficoll-Hypaque gradients. Bone marrow-derived macrophages can be produced by differentiating nonadherent macrophage precursors with medium containing macrophage colony-stimulating factor. PMID- 15361659 TI - Development of T-lymphocytes in mouse fetal thymus organ culture. AB - Fetal thymus organ culture (FTOC) is a unique and powerful culture system that allows intrathymic T-lymphocyte development in vitro. T-cell development in FTOC well represents fetal thymocyte development in vivo. Here, we describe the basic method for FTOC as well as several related techniques, including the reconstitution of thymus lobes with T-lymphoid progenitor cells, high-oxygen submersion culture, time-lapse visualization of thymic emigration, reaggregation culture, and retrovirus-mediated gene transfer to developing thymocytes in FTOC. PMID- 15361660 TI - In vitro generation of lymphocytes from embryonic stem cells. AB - Lymphocytes arise during ontogeny via a series of increasingly restricted intermediates. Initially, the mesoderm gives rise to hemangioblasts, which can differentiate into endothelial precursors, or hematopoietic stem cells (HSCs). HSCs can either self-renew or differentiate into lineage-restricted progenitors and, ultimately, to mature effector cells. This complex process is only beginning to be understood, and the ability to generate lymphocytes from embryonic stem (ES) cells in vitro will facilitate further study by providing a model system in which the effects of genetic and environmental manipulations of ES-cell-derived progenitors can be examined. In this protocol, we describe procedures for generating either B- and NK- or T-lymphocytes from mouse ES cells in vitro. PMID- 15361661 TI - Hematopoietic development of human embryonic stem cells in culture. AB - The isolation of embryonic stem (ES) cells from human preimplantation blastocysts creates an exciting new starting point to analyze the earliest stages of human blood development. This chapter describes two methods to promote hematopoietic differentiation of human ES cells: stromal cell coculture and embryoid body formation. Better understanding of basic human hematopoiesis through the study of human ES cells will likely have future therapeutic benefits. PMID- 15361662 TI - Generation of murine stromal cell lines: models for the microenvironment of the embryonic mouse aorta-gonads-mesonephros region. AB - We describe a method to derive cell lines and clones from cells of the murine midgestation aorta-gonads-mesonephros (AGM) microenvironment. We start from subdissected AGM regions in "explant-" or "single-cell suspension"-type cultures from embryos transgenic for tsA58, a temperature-sensitive mutant of the SV40 T antigen gene. The number of cells in such cultures initially expand, but in most cases, this expansion phase is followed by a stable or even decline in cell number. After this so-called crisis phase, cell proliferation is noticeable in more than 90% of the cultures. Stromal cell clones can be isolated from these cultures, some of which have been cultured for more than 50 population doublings. These stromal cell clones are valuable tools for the study of the regulation of hematopoietic stem and progenitor cells in the midgestation mouse embryo. PMID- 15361663 TI - Culture of human and mouse mesenchymal cells. AB - Normal human and mouse bone marrow is composed of hematopoietic and nonhematopoietic cells. The latter have also been termed stromal cells, microenvironment cells, colony-forming-unit fibroblasts (CFU-F), and mesenchymal cells. These cells were originally thought to provide an appropriate matrix for hematopoietic cell development, but recent examination of these cell populations suggests a much broader spectrum of activity, including the generation of bone, cartilage, muscle, tendon, and fat. In the future, these mesenchymal cell populations could be used for the treatment of specific diseases and to enhance the engraftment of hematopoietic cells. This chapter describes methods for the human CFU-F assay, culture and expansion of mesenchymal cells, as well as their differentiation to adipocytes. In addition, this chapter describes the mouse CFU F assay. PMID- 15361664 TI - Isolation, purification, and cultivation of murine and human keratinocytes. AB - The architecture of mammalian skin incorporates an outer layer of stratified epithelium. This enables the organism to conserve internal homeostasis and maintain protection from adverse environmental exposure. The keratinocyte is the cell primarily responsible for this structure. Isolation and in vitro cultivation of this cell type is widely used in dermatological and other investigations as opposed to using whole animals. However, this cell is very fastidious as compared to other skin cells (fibroblasts, etc.) and thus requires special procedures to obtain successful in vitro cultivation. This chapter describes the methodology required to isolate, purify, and cultivate keratinocytes to produce both monolayer and stratified cultures. The methodologies for producing cultures of keratinocytes obtained from rat skin and from human skin are described. PMID- 15361665 TI - Isolation and culture of primary human hepatocytes. AB - As our knowledge of the species differences in drug metabolism and drug-induced hepatotoxicity has expanded significantly, the need for human-relevant in vitro hepatic model systems has become more apparent than ever before. Human hepatocytes have become the "gold standard" for evaluating hepatic metabolism and toxicity of drugs and other xenobiotics in vitro. In addition, they are becoming utilized more extensively for many kinds of biomedical research, including a variety of biological, pharmacological, and toxicological studies. This chapter describes methods for the isolation of primary human hepatocytes from liver tissue obtained from an encapsulated end wedge removed from patients undergoing resection for removal of liver tumors or resected segments from whole livers obtained from multiorgan donors. The maintenance of normal cellular physiology and intercellular contacts in vitro is of particular importance for optimal phenotypic gene expression and response to drugs and other xenobiotics. As such, methods are described for culturing primary hepatocytes under various matrix compositions and geometries. Differential expression of liver-selective properties occurs over time in primary hepatocytes dependent on the culture and study conditions. Overall, improved isolation and cultivation methods have allowed for exciting advances in our understanding of the pathology, biochemistry, and cellular and molecular biology of human hepatocytes. PMID- 15361666 TI - Primary kidney proximal tubule cells. AB - Primary rabbit kidney epithelial cell cultures can be obtained that express renal proximal tubule functions. Toward these ends, renal proximal tubules are purified from the rabbit kidney by the method of Brendel and Meezan. To summarize, each kidney is perfused with iron oxide, which becomes associated with glomeruli. The renal cortex is sliced and homogenized to liberate nephron segments. Renal proximal tubules and glomeruli are purified by sieving. The glomeruli, covered with iron oxide, are removed using a magnet. After a brief collagenase treatment (to disrupt basement membrane), the tubules are plated in hormonally defined serum-free medium supplemented with 5 micro g/mL bovine insulin, 5 microg/mL human transferrin, and 5 x 10(-8) M hydrocortisone. After 5-6 d of incubation, confluent monolayers are obtained that possess multicellular domes, indicative of their capacity for transepithelial solute transport. PMID- 15361667 TI - Enzymatic dissociation and culture of normal human mammary tissue to detect progenitor activity. AB - Normal human mammary tissue is composed of a glandular epithelium embedded within a fibrous and fatty stroma. Collagenase and hyaluronidase digestion of normal reduction mammoplasty specimens followed by differential centrifugation yields a suspension of single cells and cell aggregates that contain elements of the terminal ductal lobular units and stromal components of the mammary gland. The terminal ductal lobular units (TDLU) can be further dissociated to complete viable single-cell suspensions by treatment with trypsin, dispase II, and deoxyribonuclease I. These suspensions are suitable for cell separation and analysis in culture. Such studies indicate the existence of biologically distinct subpopulations of luminal-restricted, myoepithelial-restricted, and bipotent mammary epithelial progenitors detected by their ability to generate colonies of the corresponding progeny types in serum-free cultures. This review summarizes the methodology of the techniques required to generate and characterize the colonies obtained in vitro from these progenitors, as well as the special considerations and potential pitfalls associated with performing these protocols. PMID- 15361668 TI - Generation and differentiation of neurospheres from murine embryonic day 14 central nervous system tissue. AB - Murine embryonic day 14 or E14 neural stem cells (NSCs), first isolated and characterized as a stem cell in culture, are a unique population of cells capable of self-renewal. In addition, they produce a large number of progeny capable of differentiating into the three primary phenotypes-neurons, astrocytes, and oligodendrocytes-found in the adult mammalian central nervous system (CNS). A defined serum-free medium supplemented with epidermal growth factor (EGF) is used to maintain the NSCs in an undifferentiated state in the form of clusters of cells, called neurospheres, for several culture passages. When EGF is removed and serum added to the medium, the intact or dissociated neurospheres differentiate into the three primary CNS phenotypes. This chapter outlines the simple NSC culture methodology and provides some of the more important details of the assay to achieve reproducible cultures. PMID- 15361669 TI - Isolation and culture of skeletal muscle myofibers as a means to analyze satellite cells. AB - Myofibers are the functional contractile units of skeletal muscle. Mononuclear satellite cells located between the basal lamina and the plasmalemma of the myofiber are the primary source of myogenic precursor cells in postnatal muscle. This chapter describes protocols used in our laboratory for isolation, culturing, and immunostaining of single myofibers from mouse skeletal muscle. The isolated myofibers are intact and retain their associated satellite cells underneath the basal lamina. The first protocol discusses myofiber isolation from the flexor digitorum brevis (FDB) muscle. Myofibers are cultured in dishes coated with Vitrogen collagen, and satellite cells remain associated with the myofibers undergoing proliferation and differentiation on the myofiber surface. The second protocol discusses the isolation of longer myofibers from the extensor digitorum longus (EDL). Different from the FDB myofibers, the longer EDL myofibers tend to tangle and break when cultured together; therefore, EDL myofibers are cultured individually. These myofibers are cultured in dishes coated with Matrigel. The satellite cells initially remain associated with the myofiber and later migrate away to its vicinity, resulting in extensive cell proliferation and differentiation. These protocols allow studies on the interplay between the myofiber and its associated satellite cells. PMID- 15361670 TI - Adult ventricular cardiomyocytes: isolation and culture. AB - Isolated cardiomyocytes are a prerequisite to study the biology of cardiomyocytes. Efficient isolation is difficult, as these cells adhere firmly together in the heart and do not divide. Therefore, any experiment is restricted to the amount of calcium-tolerant, rod-shaped cardiomyocytes that can be initially isolated from the heart. This chapter gives detailed instructions on how ventricular cardiomyocytes can be isolated from an intact adult heart. The method is based on the principle of calcium-free perfusion with collagenase supplementation to disrupt cell-cell contacts in the heart, isolation and purification of cardiomyocytes from other cell types, and, finally, re establishing a physiological cellular calcium concentration. The chapter also summarizes some commonly used adaptations to isolate cardiomyocytes from species different from rat. PMID- 15361671 TI - Isolation and culture of primary endothelial cells. AB - The purpose of this chapter is to describe the isolation techniques that result in pure cultures of human vascular endothelial cells from the umbilical vein and umbilical cord blood. We first describe the isolation of human umbilical vein endothelial cells (HUVECs). Additional protocols describe the isolation of umbilical-cord-blood-derived endothelial cells, the basic procedures of endothelial cell culture (including cryopreservation) and the methods used to characterize the phenotype of endothelial cells. PMID- 15361672 TI - Studying leukocyte rolling and adhesion in vitro under flow conditions. AB - Leukocyte recruitment from the vasculature occurs under conditions of hemodynamic shear stress. The parallel-plate flow chamber apparatus is an in vitro system that is widely used to study leukocyte recruitment under shear conditions. The flow chamber is a versatile tool for examining adhesive interactions, as it can be used to study a variety of adhesive substrates, ranging from monolayers of primary cells to isolated adhesion molecules, and a variety of adhesive particles, ranging from leukocytes in whole blood to antibody-coated Latex beads. We describe methods for studying leukocyte recruitment to cytokine-stimulated endothelial cells using both whole blood and isolated leukocyte suspensions. These methods enable multiple parameters to be measured, including the total number of recruited leukocytes, the percentage of leukocytes that are rolling or firmly adherent, and the percentage of leukocytes that have transmigrated. Although these methods are described for interactions between leukocytes and endothelial cells, they are broadly applicable to the study of interactions between many combinations of adhesive substrate and adhesive particles. PMID- 15361673 TI - Isolation and characterization of side population cells. AB - The protocol for isolation of side population (SP) cells was originally established for murine bone marrow hematopoietic stem cells (HSCs), but it has also been adapted for other species and tissues. This purification strategy offers a simple and reproducible strategy to obtain a highly homogeneous population of HSCs. The method is based on the differential efflux of the fluorescent DNA-binding dye Hoechst 33342 from stem cells relative to nonstem cells. The protocols outlined in this chapter describe the isolation of murine SP cells from both bone marrow and skeletal muscle using the fluorescent DNA-binding dye Hoechst 33342. In these tissues, the SP cells that are isolated are HSCs. PMID- 15361674 TI - Scalable production of embryonic stem cell-derived cells. AB - Embryonic stem (ES) cells have the ability to self-renew as well as differentiate into any cell type in the body. These traits make ES cells an attractive "raw material" for a variety of cell-based technologies. However, uncontrolled cell aggregation in ES cell differentiation culture inhibits cell proliferation and differentiation and thwarts the use of stirred suspension bioreactors. Encapsulation of ES cells in agarose microdrops prevents physical interaction between developing embryoid bodies (EBs) that, in turn, prevents EB agglomeration. This enables use of stirred suspension bioreactors that can generate large numbers of ES-derived cells under controlled conditions. PMID- 15361675 TI - Under- and overnutrition: a global perspective. PMID- 15361676 TI - Environmental influences on diseases in later life. PMID- 15361677 TI - Insulin sensitivity: normal and abnormal. PMID- 15361678 TI - Insulin resistance in catabolic diseases. PMID- 15361679 TI - Mechanisms of insulin-induced alterations in metabolism during critical illness. PMID- 15361680 TI - The metabolic syndrome as a clinical problem. PMID- 15361681 TI - The effect of exercise and diet on glucose intolerance and substrate utilization? PMID- 15361682 TI - Muscle mass and protein metabolism. PMID- 15361683 TI - Fatty acid isomers in lipid metabolism. PMID- 15361684 TI - Pathogenic role of inflammatory cytokines in obesity: from insulin resistance to diabetes mellitus. PMID- 15361685 TI - Diet modifications to control hypertension: the multifactorial approach. PMID- 15361686 TI - The role of minerals and trace elements in relation to long-term health and chronic disease. PMID- 15361687 TI - Fluid and electrolytes in the clinical setting. PMID- 15361688 TI - Water and sodium balance: a nutritional goal. PMID- 15361689 TI - Neuro-hormonal integration of metabolism: challenges and opportunities in the postgenomic era. PMID- 15361690 TI - Nutrition and genomics. PMID- 15361691 TI - Hexarelin modulates the expression of growth hormone secretagogue receptor type 1a mRNA at hypothalamic and pituitary sites. AB - Ghrelin and the synthetic growth hormone secretagogues (GHSs) activate a G protein-coupled receptor (GHS-R) originally cloned from the pituitary, but which is also expressed in the hypothalamus, in other areas of the brain and in numerous peripheral tissues. Several studies have shown that growth hormone (GH) releasing hormone (GHRH) is necessary for GHSs to exert maximal GH release in vivo. The exact mechanism of this synergism is not clear. Previous data suggest that GHSs can affect pituitary GHS-R mRNA expression; however, it is unknown whether this effect is age dependent and whether hypothalamic GHS-Rs are also affected. In this study, we tested whether (a) the synthetic GHS hexarelin regulates mRNA expression of its own receptor at the pituitary and/or hypothalamus and whether this effect is age dependent, and (b) whether short-term treatment with GHRH or, conversely, passive immunization against GHRH affects pituitary GHS-R1a mRNA expression in infant (10 days old) and young adult rats. GHS-R1a mRNA expression was measured with competitive reverse transcriptase polymerase chain reaction. Hexarelin treatment significantly increased pituitary and hypothalamic GHS-R1a mRNA levels in normal infant rats, but not in normal young adult rats. In addition, hexarelin administration also stimulated pituitary GHS-R1a mRNA in infant as well as in young adult rats passively immunized against GHRH. GHRH treatment significantly enhanced pituitary GHS-R1a mRNA expression in GHRH-deprived young adult rats, though it did not affect the basal levels of GHS R1a mRNA in normal infant and adult rats. These data further support the hypothesis that GHRH can affect GHS-R1a expression and that hexarelin upregulates the expression of its own receptor at the pituitary as well as the hypothalamus in an age-dependent fashion. PMID- 15361692 TI - Therapies to slow polycystic kidney disease. AB - Advances in the understanding of cystogenesis and availability of animal models orthologous to human autosomal dominant polycystic kidney disease (ADPKD) and recessive polycystic kidney disease (ARPKD) will likely facilitate the development of treatments for these diseases. Proteins mutated in ADPKD and ARPKD, as well as in several animal models, are localized to renal primary cilia. These are thought to have a sensory function and contribute to the regulation of the intracellular calcium ([Ca2+]i). It seems likely that the maintenance of a differentiated renal epithelial phenotype, characterized by controlled fluid secretion and cell proliferation, requires precise functional coordination of cAMP and Ras/Raf/MEK/ERK signaling by [Ca2+]i. [Ca2+]i alterations, linked to genetic defects causing polycystic kidney disease, may hinder negative feedback mechanisms that control cAMP and Ras/Raf/MEK/ERK signaling, and result in increased fluid secretion and cell proliferation. cAMP levels, Raf kinase activities and ERK phosphorylation are increased in polycystic kidneys. There is also evidence of abnormal cross-talk between cAMP and MAPK pathways, that can be reproduced in wild-type cells by altering [Ca2+]i. While cAMP inhibits Ras-Raf-1 stimulated phosphorylation of ERK in normal kidney cells, it markedly increases B Raf kinase activity and ERK phosphorylation in polycystic kidney cells. Treatment strategies should probably be aimed at increasing [Ca2+]i, inhibiting Ras/Raf/MEK/ERK signaling or lowering cAMP in the distal nephron and collecting duct. Vasopressin is the major adenylyl cyclase agonist in the collecting duct principal cells via a V2 receptor. OPC31260, a V2 receptor antagonist, lowers renal cAMP and markedly inhibits cystogenesis in four animal models of polycystic kidney disease, three of which are orthologous to human diseases (PCK rat, ARPKD; pcy mouse, adolescent nephronophthisis; Pkd2WS25/- mouse, ADPKD). The renal selectivity and safety profile of this class of drugs make it an excellent candidate for clinical trials. PMID- 15361693 TI - Hypoxia and tubulointerstitial injury: a final common pathway to end-stage renal failure. AB - Many clinical observations suggest common mediators in the progression of kidney disease leading to eventual kidney failure. Among them, accumulating evidence emphasizes the role of chronic hypoxia in the tubulointerstitium in this role. When advanced, tubulointerstitial damage is associated with the loss of peritubular capillaries, impairing blood delivery. Associated interstitial fibrosis further impairs oxygen diffusion and supply to tubular and interstitial cells. This in turn exacerbates chronic hypoxia in this compartment, resulting in a vicious cycle. Both singly or together, glomerular injury and vasoconstriction of efferent arterioles due to an imbalance in vasoactive substances decrease post glomerular peritubular capillary blood flow and contribute to chronic hypoxia in the tubulointerstitium. Anemia in kidney disease also plays a significant role in hypoxia of the kidney. Moreover, increased metabolic demand in tubular cells, as observed in glomerular hyperfiltration for example, can cause relative hypoxia. Importantly, these factors can affect the kidney before the appearance of significant pathological changes in the vasculature and predispose it to tubulointerstitial injury. Therapeutic approaches targeting chronic hypoxia in the kidney should be effective against a broad range of renal diseases. Recent studies have elucidated the mechanisms of hypoxia-induced transcription, giving hope for the development of novel therapeutic approaches against this final common pathway. PMID- 15361694 TI - Aggregated IgG bind to glomerular epithelial cells to stimulate urokinase release through an endocytosis-independent process. AB - BACKGROUND: In membranous nephropathy, the development of glomerular lesions is related to the formation of immune complexes at subepithelial sites. These deposits are associated with modifications in the fibrinolytic activity of glomerular cells leading to the appearance of fibrin degradation products in the deposits and the urine. A previous study has shown that immune complexes interact with glomerular epithelial cells (GEC) through the neonatal Fc receptor (FcRn). We therefore determined whether this binding could be responsible for a modification in the fibrinolytic activity of GEC. METHODS: Endocytosis of heat aggregated immunoglobulins (AgIgG) in cultured human GEC was studied by immunofluorescence and confocal microscopy. The release of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor 1 (PAI) by GEC or whole glomeruli was assessed by ELISA, fibrin zymography and Northern blot. RESULTS: Human GEC in culture bound AgIgG that possess characteristics similar to soluble immune complexes and internalized them by 10 min. This process was mediated by FcRn since chicken aggregated IgG (AgIgY), that do not bind FcRn, did not colocalize with AgIgG in GEC. AgIgG but not AgIgY induced a decrease of FcRn expression at the membrane and within the cells. The binding of AgIgG to GEC elicited a dose- and time-dependent increase in the release of uPA activity, as in the uPA protein and mRNA expression without modification in the release of PAI. This process was not abrogated by agents inhibiting endocytosis and/or transcytosis such as cytochalasin B, suggesting an endocytosis-independent uPA regulation. CONCLUSION: GEC response to AgIgG overload comprises at least two sequential steps: (1) a FcRn-mediated endocytosis; (2) an endocytosis-independent fibrinolytic imbalance leading to plasmin generation which could favor in vivo AgIgG clearance and matrix remodeling. PMID- 15361695 TI - Redistribution of integrins in tubular epithelial cells during diabetic glycogen nephrosis. AB - BACKGROUND/AIMS: Even though many aspects of glycogen nephrosis in diabetes have already been studied, adhesion interactions between the glycogen-accumulating clear cells and the tubular basement membranes have not been addressed. As integrins play key roles in cell-to-matrix interactions, we investigated the expression and distribution of alpha3-, alphaV-, beta1- and beta3-integrin subunits in renal tissues from streptozotocin-induced hyperglycemic rats (3 months old) and their age-matched controls as well as from streptozotocin injected normoglycemic animals. METHODS: The levels and distribution of integrins were studied by immunocytochemistry and Western blot analysis. RESULTS: Immunoblotting analysis of fractions enriched in glycogen-accumulating clear cells demonstrated enhanced expression of alpha3, alphaV and beta1 subunits while expression of beta3 did not differ from controls. The most striking cytochemical result was the redistribution of the alpha3-, alphaV-, and the beta1-integrin subunits to the apical plasma membrane of these cells. This was found by light and electron microscopy. CONCLUSION: Our results suggest that the altered expression and distribution of integrins in clear cells of diabetic animals must have defined roles in the development of the renal tubulopathy. PMID- 15361696 TI - Role of the B7RP-1/ICOS pathway in renal tubular epithelial antigen presentation to CD4+ Th1 and Th2 cells. AB - BACKGROUND: IFN-gamma-stimulated renal tubular epithelial cells (TECs) can present foreign protein antigen to T cells when stimulated with IFN-gamma. TECs normally do not express B7, the ligand for the classical T-cell costimulatory molecule CD28, but express B7RP-1, the ligand of the inducible costimulator (ICOS). METHODS: Flow cytometry was used to demonstrate expression of B7RP-1 on TECs, and expression of ICOS on A.E7 (Th1 clone) and D10.G4 (Th2 clone) cells. Using gene arrays, ELISA, and protein arrays, we examined the cytokine response of Th1 and Th2 subsets of CD4+ T cells in antigen presentation by IFN-gamma stimulated murine primary TECs. RESULTS: In the presence of a specific foreign protein antigen, MHC-II-positive TECs induced clustering and a marked production of IFN-gamma (A.E7) or IL-4 (D10.G4) by T cells. Blockade of the B7RP-1/ICOS pathway with specific monoclonal antibodies (mAbs) lead to a further increase in IFN-gamma production by A.E7 cells, and a further increase in IL-4 production by D10.G4 cells as detected by gene array and ELISA. Protein arrays demonstrated an increase in Th2 cytokine levels upon blockade of the B7RP-1/ICOS pathway. CONCLUSIONS: These data show that the B7RP-1/ICOS interaction inhibits Th1 and Th2 T-cell responses in the setting of foreign antigen presentation by renal TECs. This inhibitory pathway may therefore serve as a negative feedback mechanism to inhibit immune-mediated tubulointerstitial renal disease. PMID- 15361697 TI - Which marker is informative in characterizing the level of oxidative stress in ESRD patients? PMID- 15361698 TI - Oxidative stress in patients with end-stage renal disease prior to the start of renal replacement therapy. AB - BACKGROUND/AIM: In patients with end-stage renal disease (ESRD), cardiovascular complications are the main cause of death. Increased oxidative stress is one of the risk factors for enhanced atherosclerosis in this population. Literature data vary partially dependent on differences in methodology. The present study compares three different methods: plasma lipid peroxides, the newly developed measurement of circulating oxidized LDL (Ox-LDL) particles and the frequently used copper-induced LDL oxidation lag time. METHODS: We assessed plasma lipid peroxides, circulating Ox-LDL and in vitro copper-induced LDL oxidation lag time in 47 non-diabetic patients with ESRD, at the start of renal replacement therapy, and compared these with 41 age- and sex-matched controls. RESULTS: In ESRD, total cholesterol (4.6 +/- 1.1 vs. 5.6 +/- 0.9 mmol/l; p < 0.001), LDL cholesterol (2.8 +/- 0.8 vs. 3.5 +/- 0.7 mmol/l; p < 0.001) and HDL cholesterol (1.0 +/- 0.3 vs. 1.4 +/- 0.4 mmol/l; p < 0.001) were lower compared to controls. Plasma lipid peroxides were higher (1.1 +/- 0.5 vs. 0.8 +/- 0.5 micromol/l; p = 0.003) in ESRD. No differences were observed in plasma Ox-LDL (63.1 +/- 62.0 vs. 55.3 +/- 48.0 mg/l). However, due to the lower plasma LDL cholesterol in ESRD, LDL oxidation level was increased in ESRD (7.1 +/- 0.1 vs. 4.2 +/- 0.3%; p = 0.03). LDL lag time was slightly longer (89 +/- 11 vs. 84 +/- 11 min; p = 0.04) in ESRD. There were no significant differences regarding the amount and rate of dienes produced. CONCLUSIONS: Elevated levels of lipid peroxides and higher LDL oxidation levels support the theory that ESRD is associated with increased oxidative stress, which may explain the accelerated atherosclerosis. The measured amount of oxidative stress is not reflected by in vitro oxidizability of LDL. PMID- 15361699 TI - Pattern of renal function recovery after leptospirosis acute renal failure. AB - BACKGROUND: Although acute renal failure (ARF) is a frequent complication of severe leptospirosis, there are few studies on renal function recovery in the literature. The objective of the present study was to verify how and when renal function recovery occurs after leptospirosis ARF. PATIENTS AND METHODS: 35 patients with leptospirosis ARF (plasma creatinine [P(creat)] > or =133 micromol/l at hospital admission) were prospectively studied during hospitalization, at discharge and 3 and 6 months after discharge. Creatinine clearance, fractional excretion of sodium and potassium, proteinuria and sodium proximal reabsorption were measured under normal sodium diet. Urinary pH and the ratio urinary to plasma osmolality (U/P(osm)) were measured 18 h after food and water withdrawal. All parameters were also measured in 18 healthy volunteers. Presence of laboratorial alterations usually found in leptospirosis were evaluated in the patients. Patients were divided in 2 groups according to their maximum P(creat) value during hospitalization: group 1 ( P(creat) >442 micromol/l, n = 21), and group 2 (P(creat) < or =442 micromol/l, n = 14). RESULTS: All patients presented with jaundice, fever and myalgia. Bilirubin and creatine kinase were higher in group 1. Oliguria was observed in 11% of all patients and 49% required dialysis; all these patients were from group 1. All renal parameters were normal at the 6th month except U/P(osm) that remained lower than normal. The pattern of renal function recovery was similar in both groups except for urinary volume. CONCLUSION: After leptospirosis ARF, renal function recovery is fast and complete after 6 months, except for urinary concentration capacity. PMID- 15361700 TI - Clinical significance of serum oxidized low-density lipoprotein/beta2 glycoprotein I complexes in patients with chronic renal diseases. AB - BACKGROUND: Peroxidation of low-density lipoprotein (LDL) plays an important role in the development of dyslipidemias associated with the progression of chronic renal disorders. We recently reported [J Lipid Res 2001;42:697, 2002;43:1486, 2003;44:716] that oxidized LDL (oxLDL) interacts with an endogenous plasma protein, beta2-glycoprotein I (beta2GPI), via specific ligands. In the present study, the prevalence and clinical significance of oxLDL/beta2GPI complexes were evaluated in patients with chronic renal disorders. METHODS: Serum levels of oxLDL/beta(2)GPI complexes were measured by ELISA in patients with chronic renal disease and their association with clinical manifestations was assessed. RESULTS: The serum levels of oxLDL/beta2GPI complexes were significantly higher in patients with chronic renal failure (CRF), chronic nephritis (CN) and diabetes mellitus than those in healthy individuals. The presence of complexes in patients with CN was significantly associated with high dietary protein and sodium chloride intake, but not with lipid metabolic parameters. Malondialdehyde modified LDL was significantly associated with total cholesterol and LDL cholesterol in all patient groups, but did not correlate with renal function parameters. CONCLUSIONS: Serum oxLDL/beta2GPI complexes, generated by oxidative stress and associated with high dietary protein and salt intake, might be a novel risk factor and a diagnostic marker for the development of chronic renal diseases, especially IgA nephropathy. PMID- 15361701 TI - CD68 and MCP-1/CCR2 expression of initial biopsies reflect the outcomes of membranous nephropathy. AB - BACKGROUND: A considerable diversity of prognosis is seen with idiopathic membranous nephropathy (IMN). The initial factors affecting long-term outcome remain unclear. METHODS: We studied retrospectively 30 patients with IMN who had been followed up for at least 5 years, or until end-stage renal failure (ESRF). We analyzed the prognostic factors of ESRF in the first renal biopsies; these factors included presence of tubulointerstitial lesions and foam cells, as well as expression of CD68, monocyte chemoattractant protein-1 (MCP-1), its cognate receptor chemokine receptor 2 (CCR2) and alpha-smooth muscle actin. RESULTS: The patients who developed ESRF showed higher percentages of glomerular segmental sclerosis, interstitial MCP-1 expression, CCR2- or CD68-positive monocyte/macrophages and foam cells in the interstitium, and these proved on multivariate analysis to be independent risk factors for ESRF. Finally, ESRF was characterized by the presence of ten interstitial CD68-positive cells per visual field at 200x magnification (hazard ratio 4.096, CI 1.271-15.029, p < 0.001). CONCLUSIONS: Our results suggested that an interstitial infiltration of CD68 positive cells accompanied by MCP-1/CCR2 expression is the most significant indicator of ESRF in IMN. PMID- 15361702 TI - A simple, safe and effective citrate anticoagulation protocol for the genius dialysis system in acute renal failure. AB - BACKGROUND: The Genius dialysis system is a close loop dialysis batch system increasingly used as an intermittent hemodialysis device in intensive care units. The aim of this study was to test the safety and feasibility of a regional citrate anticoagulation protocol with respect to acid-base and electrolyte disarrangements in critically ill patients with acute renal failure. A standard heparin anticoagulation protocol served as control. METHODS AND RESULTS: In a cross-over study design, 27 acute renal failure patients were allocated to a citrate- and heparin-anticoagulated dialysis sessions (4-6 h). For citrate anticoagulation, a 4% sodium-citrate solution was infused into the arterial line of the extracorporeal circuit. A low calcium dialysate (1 mmol/l) was used for all dialysis sessions. Citrate dosing was adjusted according to the post-filter ionized calcium concentration (targeted values 0.5-0.7 mmol/l). There was no routine calcium substitution. Heparin anticoagulation was started with a heparin loading dose followed by an individual, patient-adjusted continuous heparin infusion. Electrolyte disarrangements, namely hypernatremia, hypo- and hypercalcemia did not occur in either group. Although the highest bicarbonate levels were achieved during citrate anticoagulation (p = 0.021 versus heparin) the acid base values remained equilibrated in both groups. Filter longevity was excellent and the targeted dialysis time was achieved in all but 1 patient. Citrate anticoagulation was well tolerated with respect to cardiovascular hemodynamics. CONCLUSIONS: Citrate anticoagulation can be safely and effectively performed during intermittent Genius dialysis. Calcium supplementation is not routinely required. PMID- 15361703 TI - Female age in ART: when to stop? AB - With increasing age the probability of ongoing pregnancy established by the use of assisted reproduction technology (ART) decreases. As a result the question arises whether age limits for the application of ART should be established. From a literature review and ongoing research data it appears that the costs per child born greatly increase after the age of 40 for both intrauterine insemination with mild ovarian stimulation and in vitro fertilisation treatment, while in cases of 44 and over, prognosis is flat zero. The willingness to pay for extra costs will greatly determine whether and at what age strict limits should be applied. Fortunately, predictive factors for success, like the antral follicle count, may enable the identification of women over 40 and under 44 that still have favourable prospects, thereby decreasing the necessary costs per childbirth and allowing couples into ART programs that are often denied based solely on female age. PMID- 15361704 TI - Evaluation of vitamin B6 status and RDA in young Koreans. AB - AIMS AND METHODS: The vitamin B6 status of 294 healthy young Koreans (149 men, 145 women), aged 17-25 years, was estimated to establish the Korean Recommended Dietary Allowance (RDA) for vitamin B6. The dietary vitamin B6 intake was estimated based on a three-day recall method. RESULTS: The average daily vitamin B6 intake of the subjects was 1.504 +/- 0.825 mg/day (1.567 +/- 0.968 mg/day for men and 1.439 +/- 0.644 mg/day for women). The average ratio of vitamin B6 intake to daily protein intake was 0.019 +/- 0.001 mg/g (0.015 +/- 0.001 mg/g for men and 0.023 +/- 0.001 mg/g for women). Foods from animal and plant sources provided 35.68 +/- 16.16 and 64.18 +/- 16.29% of total vitamin B6. Plasma pyridoxal 5' phosphate (PLP) concentration of the subjects was 29.26 +/- 16.84 nmol/l (30.52 +/- 20.18 nmol/l for men and 28.21 +/- 13.28 nmol/l for women). The vitamin B6 intake of the subjects whose PLP concentration was >30 nmol/l was 0.019 +/- 0.001 mg vitamin B6/g protein intake (0.017 +/- 0.001 and 0.023 +/- 0.001 mg vitamin B6/g protein intake for men and women, respectively). The intake of vitamin B6 was significantly (p < 0.01-p < 0.001) positively correlated with the intake of all other nutrients. However, no significant correlation was found between PLP concentration and all nutrients examined. CONCLUSION: Our suggested adequate Korean RDA for vitamin B6 in young Koreans is 1.6 mg/day or 0.02 mg/g of protein intake in men and 1.5 mg/day or 0.02 mg/g of protein intake in women. PMID- 15361705 TI - Expression failure of the notch signaling system is associated with the pathogenesis of testicular germ cell tumor. AB - OBJECTIVES: The expression of Notch 1 and its ligand Jagged 2, critical factors in cell type specification, in testicular germ cell tumors was examined in order to evaluate its possible relationship with their pathogenesis. METHODS: Northern blot analysis and immunohistochemical staining for Notch 1 and Jagged 2 were done in 139 samples of testicular germ cell tumors. RESULTS: Notch 1 and Jagged 2 transcripts were expressed in noncancerous testicular tissues and Notch 1 and Jagged 2 proteins were positive in the spermatids. However, the expressions of transcript and protein were negative for both Jagged 2 and Notch 1 in seminomas, they were negative for Jagged 2 but were positive for Notch 1 in embryonal carcinomas and choriocarcinomas, and they were positive for Jagged 2 and Notch 1 in teratomas. CONCLUSIONS: Our results offer, for the first time, the possibility that the activity of the Notch signaling system, one of the cell proliferation and differentiation pathways, correlates with the different histological subtypes of germ cell tumor, which may be responsible for characteristics of cancer cells such as responses to chemotherapy and/or irradiation. PMID- 15361706 TI - Predictive and prognostic role of E-cadherin protein expression in patients with advanced gastric carcinomas treated with palliative chemotherapy. AB - Loss of E-cadherin expression has been related with an adverse outcome in patients with resected gastric cancer. More recently, experimental models with cancer cell lines showed that chemosensitivity may be affected by the E-cadherin expression status. We investigated whether E-cadherin expression is correlated with the response to chemotherapy and the survival of patients with advanced gastric cancer. Consecutive patients with advanced gastric cancer who underwent palliative chemotherapy were considered eligible for study entry. Measurable disease, complete follow-up information and availability of tumor specimens for immunohistochemistry were mandatory inclusion criteria. In 70 assessable patients, 30 patients had locoregional disease and 40 patients had visceral metastases. Chemotherapy consisted of cisplatin/fluorouracil/folinic acid in 33 patients and cisplatin/fluorouracil/epirubicin/folinic acid in 37 patients. There were 13 patients with complete response, 20 with partial response, 20 with stable disease and 17 patients progressed. Thirty-eight patients had > 80% E-cadherin positive cancer cells (positive E-cadherin expression); 15 cases showed 25-70% (reduced E-cadherin expression), and in the remaining 17 cases E-cadherin expression was < 10% (negative E-cadherin expression). The response to chemotherapy was unrelated to the E-cadherin expression status. Conversely, survival in the 32 patients with reduced/negative E-cadherin expression (25 weeks) was significantly worse than that observed in the 38 patients with preserved E-cadherin expression (36 weeks) (p < 0.01). E-cadherin expression retained its independent prognostic role in the multivariate analysis. E-cadherin expression may give prognostic information in patients with advanced gastric cancer, but it does not seem to possess a predictive role in vivo. Some of the mechanisms inducing E-cadherin downregulation, like hypermethylation, may be potentially reversible, and they deserve further investigation as the target of novel therapeutic strategies. PMID- 15361707 TI - Plasma levels of cellular fibronectin in patients with localized and metastatic renal cell carcinoma. AB - Clinical markers for renal cell carcinoma (RCC) are lacking. Fibronectin is a glycoprotein that plays an important role in cellular attachment and cell spread. The aim of this study was to test the clinical suitability of cellular fibronectin (cFN) in plasma as a tumor marker for RCC and to determine a possible relationship between cFN plasma levels and stage of disease. Therefore, cFN was determined in the plasma of patients with localized (n = 40) and metastatic (n = 20) RCC using a time-resolved fluorescence immunoassay. Fifty patients with different non-malignant urological disorders were recruited as a control group. In the control group, mean cFN plasma levels amounted to 553 ng/ml. In patients with localized RCC, plasma concentrations of cFN were increased (1,295 ng/ml; p < 0.01). Patients with metastatic disease had the highest concentrations (3,842 ng/ml). Statistical analysis demonstrated a significant difference between controls, and patients with localized and metastatic RCC (p < 0.01), with a sensitivity of 80%, a specificity of 78% and a positive predictive value of 81% using a cutoff value of 540 ng/ml (receiver-operating characteristic curve analysis). These data suggest that cFN is not a realistic marker for the detection of RCC. However, elevated plasma levels in more advanced disease and an acceptable predictive value could indicate that cFN is useful as a follow-up tool in the management of RCC patients. PMID- 15361708 TI - Gene expression levels of cytokines in peritoneal washings from patients with gastric cancer. AB - Peritoneal seeding is frequently detected in patients with gastric cancer. The peritoneal cavity is a compartment in which the immunologic host-tumor interaction can occur. Here, we investigated the gene expression levels of cytokines, and compared these gene expressions with the progression of gastric cancer. Total RNA was extracted from 50 ml of peritoneal washings from 78 patients with gastric cancer and 11 noncancerous patients. The gene expression levels of transforming growth factor-beta (TGF-beta), interleukin (IL)-2, IL-6 and IL-12 were analyzed by real-time reverse transcription-polymerase chain reaction. The gene expression levels of TGF-beta and IL-12 in 16 patients with peritoneal seeding (peritoneal metastasis or free cancer cells) did not differ from those in controls (n = 11) and stage I and II (n = 43) patients. However, the relative gene expression level of IL-2 in patients with peritoneal seeding (0.9) was lower than that in controls (1.4, p = 0.066) and stage I and II patients (1.4, p = 0.036). In contrast, the relative gene expression level of IL 6 in patients with peritoneal seeding (3.3) was higher than that in control (2.4, p = 0.064) and stage I and II patients (2.6, p = 0.065). Low IL-2 gene and high IL-6 gene expressions in the peritoneal cavity may correlate with cancer development in the peritoneal cavity in patients with gastric cancer. PMID- 15361709 TI - Molecular cloning of a new gene which is differentially expressed in breast and prostate cancers. AB - OBJECTIVE: The chromosomal region 19q13 is non-randomly rearranged in many solid tumors. METHODS: Using the positional candidate gene approach, we cloned a new gene, tentatively named cancer-associated gene (CAG), which is differentially expressed in breast and prostate cancers. RESULTS: The gene is formed of 3 exons and 2 intervening introns. Its coding region is 1,047 bp in length and is predicted to encode a 348-amino-acid polypeptide. The new gene maps to chromosome 19q13.4 and is located 14 kb telomeric to the kallikrein gene locus (KLK14 gene) and 17 kb centromeric from the Siglec family of genes (Siglec-9). The gene is expressed in a wide variety of tissues including the brain, colon, kidney and pancreas. The CAG protein shows a high degree of conservation among species and phylogenetically is most closely related to its mouse ortholog. In silico analysis indicates that this gene is differentially expressed in a variety of tumors including brain, colon, ovarian and prostate cancers. CONCLUSIONS: Our preliminary experimental data show that CAG is upregulated in prostate cancer tissues compared to normal prostatic tissues. CAG also appears to be downregulated in breast cancer tissues. The physiological function of the CAG protein is currently unknown. PMID- 15361710 TI - Aberrant methylation and histone deacetylation associated with silencing of SLC5A8 in gastric cancer. AB - Aberrant methylation of a sodium co-transporter, solute carrier family 5 member 8 gene (SLC5A8), has been detected in a subset of colorectal cancers, suggesting SLC5A8 may also serve as a tumor suppressor. To further investigate the role of epigenetic inactivation of SLC5A8 expression in gastric cancer, we determined the methylation status of the SLC5A8 5' CpG island (CGI) in a panel of gastric cancer cell lines and primary gastric cancers. We detected methylation of the 5'CGI in ten of twelve gastric cancer cell lines, and five of those showed dense methylation, which correlated with the absence of SLC5A8 transcription. Aberrant methylation of SLC5A8 was also detected in 23 of 71 (30%) primary gastric cancers, indicating that epigenetic inactivation of SLC5A8 is not a cell-line specific phenomenon. SLC5A8 expression was restored in methylated cell lines by treatment with 5-aza-2'-deoxycytidine, a methyltransferase inhibitor. In addition, chromatin immunoprecipitation assays showed that acetylation of histone H3 in the 5' region of the gene correlated directly with SLC5A8 expression and inversely with DNA methylation. It thus appears that aberrant methylation of its 5'CGI and histone deacetylation play key roles in silencing SLC5A8 expression in gastric cancers. PMID- 15361711 TI - The transmembrane protease serine (TMPRSS3/TADG-12) D variant: a potential candidate for diagnosis and therapeutic intervention in ovarian cancer. AB - The purpose of this study was to examine the expression of splice variants of the TADG-12 (TMPRSS3) gene in normal ovarian epithelial tissue and ovarian carcinoma and further to associate the expression of TADG-12 variant with clinicopathologic characteristics if such an association exists. TADG-12D variant expression was examined by semiquantitative PCR in 50 ovarian tumors [41 adenocarcinomas, 3 low malignant potential (LMP) tumors, and 6 adenomas] and 7 normal ovaries. In carcinomas as well as LMP tumors and adenomas, TADG-12D variant mRNA expression was significantly elevated compared to that in normal ovary samples. TADG-12 has several splice variants, one of which we originally identified and 3 others identified by Scott et al. [Nat Genet 2001;27:59-63]. We previously examined the expression of TADG-12V variant and here we confirm the overexpression of TADG-12D variant in ovarian carcinomas. Moreover, TADG-12D variant mRNA expression level in carcinomas was significantly elevated compared to that in adenomas and TADG 12D variant mRNA expression level in advanced clinical stage diseases was significantly higher than that in early stage diseases in ovarian carcinomas. With regard to histological type, TADG-12D variant mRNA expression level in mucinous adenocarcinomas was significantly higher than those in the other tissue subtypes. These features imply that TADG-12D variant expression may play an important role in ovarian cancer development and progression, and this variant may be useful both as a molecular target for therapy and/or a diagnostic marker. PMID- 15361712 TI - Differential expression of a human kallikrein 5 (KLK5) splice variant in ovarian and prostate cancer. AB - The presence of more than one mRNA form is common among kallikrein genes. We identified an mRNA transcript of the human kallikrein gene 5 (KLK5), denoted KLK5 splice variant 1 (KLK5-SV1). This variant has a different 5'-splice site, but encodes the same protein as the classical KLK5 transcript. RT-PCR analysis of this variant transcript expression in 29 human tissues indicated highest expression in the cervix, salivary gland, kidney, mammary gland, and skin. Comparative analysis of the expression levels of KLK5-SV1, another splice variant named KLK5 splice variant 2 (KLK5-SV2), and the classical KLK5 form showed that out of all three mRNA transcripts, the classical form is predominantly expressed (found in more tissues and at higher expression levels) followed by KLK5-SV1. KLK5-SV1 is expressed at high levels in ovarian, pancreatic, breast and prostate cancer cell lines. KLK5-SV1 was also found to be expressed in 9/10 ovarian cancer tissues, but it was not found in one normal ovarian tissue tested. Hormonal regulation experiments suggest that KLK5-SV1 is regulated by steroid hormones in the BT-474 breast cancer cell line. Furthermore, this variant had significantly higher expression in normal prostate tissues compared to their matched cancer tissue counterparts. KLK5-SV1 may have clinical utility in various malignancies and should be further explored as a potential new biomarker for prostate and ovarian cancer. PMID- 15361716 TI - Walter A. Orenstein, MD: a tribute from grateful pediatric and public health communities. PMID- 15361717 TI - The Pediatric Investigators Collaborative Network on Infections in Canada study of predictors of hospitalization for respiratory syncytial virus infection for infants born at 33 through 35 completed weeks of gestation. AB - BACKGROUND: Infants born at 33 through 35 completed weeks of gestation (33-35GA) are at risk for severe respiratory syncytial virus (RSV) infection, and palivizumab prophylaxis lowers hospitalizations for RSV infection by as much as 80%. The 33-35GA cohort comprises 3-5% of annual births; thus expert panels recommend limiting prophylaxis to situations in which frequency or health care impact of RSV infection is high. This study sought to identify independent risk factors for hospitalization for RSV infection. METHODS: This was a multicenter, prospective, observational cohort study of 33-35GA infants followed through their first RSV season (2001/2002 or 2002/2003). Baseline data were collected by interview with parents and review of medical records. Respiratory tract illnesses were identified by monthly phone calls, and medical records were reviewed for emergency room visits or hospitalizations. Risk factors were determined by stepwise logistic regression. RESULTS: Of 1,860 enrolled subjects, 1,832 (98.5%) were followed for at least 1 month, and 1,760 (94.6%) completed all follow-ups. Of 140 (7.6%) subjects hospitalized for respiratory tract illnesses, 66 infants had proven RSV infection. Independent predictors for hospitalization for RSV infection were: day-care attendance (odds ratio, 12.32; 95% confidence interval, 2.56, 59.34); November through January birth (odds ratio, 4.89; 95% confidence interval, 2.57, 9.29); preschool age sibling(s) (odds ratio, 2.76; 95% confidence interval, 1.51, 5.03); birth weight <10th percentile (odds ratio, 2.19; 95% confidence interval, 1.14, 4.22); male gender (odds ratio, 1.91; 95% confidence interval, 1.10, 3.31); > or = 2 smokers in the home (odds ratio, 1.87; 95% confidence interval, 1.07, 3.26); and households with >5 people, counting the subject (odds ratio, 1.79; 95% confidence interval, 1.02, 3.16). Family history of eczema (odds ratio, 0.42; 95% confidence interval, 0.18, 0.996) was protective. CONCLUSIONS: Specific host/environmental factors can be used to identify which 33-35GA infants are at greatest risk of hospitalization for RSV infection and likely to benefit from palivizumab prophylaxis. PMID- 15361718 TI - Case-control study of the risk factors linked to respiratory syncytial virus infection requiring hospitalization in premature infants born at a gestational age of 33-35 weeks in Spain. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to identify those risk factors most likely to lead to the development of RSV-related respiratory Infection and subsequent hospital admission among premature infants born at 33-35 WGA (FLIP study) METHODS: This was a prospective case-control study. Cases (186) hospitalized for respiratory syncytial virus (RSV) illness were recruited from 50 participating Spanish hospitals during the 2002-2003 RSV season (October 2002 April 2003). Controls (371) were selected in June 2003 but born at same time as cases. RESULTS: Of these cases, 20.5% were admitted to the intensive care unit intensive care unit, and 7.6% required mechanical ventilation. None of the patients died. Conditional logistic regression analysis adjusted for medical center demonstrated that the risk of RSV-related respiratory infection requiring hospital admission in preterm infants 33-35 weeks of gestation (WGA) in Spain was most often associated with absolute chronologic age at start of RSV season < or =10 weeks [ie, born between July 15 and December 15; odds ratio (OR), 3.95; 95% confidence interval (CI), 2.65-5.90], breast-feeding for < or =2 months total (OR 3.26; 95% CI 1.96-5.42), presence of > or =1 school age siblings (OR 2.85; 95% CI 1.88-4.33), > or =4 residents and visitors at home (discounting school age siblings and the case/control him/herself) (OR 1.91; 95% CI 1.19-3.07) and a family history of wheezing (OR 1.90; 95% CI 1.19-3.01). CONCLUSIONS: In premature infants born 33-35 WGA, certain underlying risk factors significantly increase the risk of RSV-related respiratory infection and hospitalization. Premature infants 33-35 WGA with additional risk factors should be considered for RSV prophylaxis with palivizumab. PMID- 15361719 TI - The unresolved issue of risk factors for hospitalization of infants with respiratory syncytial virus infection born after 33-35 weeks gestation. PMID- 15361720 TI - Changes in frequency and pathogens causing acute otitis media in 1995-2003. AB - OBJECTIVE: This study was undertaken to determine whether a change in the frequency or distribution of the causative pathogens in persistent acute otitis media (AOM) and AOM treatment failure (AOMTF) occurred after publication of the Centers for Disease Control and Prevention AOM treatment guidelines advocating high dose amoxicillin in 1998 and the universal use of the pneumococcal conjugate vaccine in 2000. METHODS: This was a 9-year prospective study in a suburban, community-based private practice. To identify bacterial isolate(s), 551 children with AOM who had not responded after 1 or 2 empiric antimicrobial treatment courses (termed persistent AOM) and those who were failures after 48 h on treatment (AOMTF) underwent tympanocentesis. Three periods were compared: (1) 1995-1997 when all enrolled received standard dose amoxicillin (40-50 mg/kg/day divided 3 times daily) as the initial empiric treatment; (2) 1998-2000 when all received high dose amoxicillin (80-100 mg/kg/day divided twice daily); and (3) 2001-2003 when high dose amoxicillin and pneumococcal conjugate vaccinations were used. RESULTS: Persistent AOM or AOMTF for which tympanocentesis was performed occurred in 195 (16.2%) of 1,207, 204 (16.1%) of 1,278 and 152 (12.3%) of 1,232 AOM visits for 1995-1997, 1998-2000 and 2001-2003, respectively; the 24% decline in 2001-2003 in persistent AOM and AOMTF was significant (P = 0.007). Middle ear aspirates grew Streptococcus pneumoniae (48, 44 and 31%) and Haemophilus influenzae (38, 43 and 57%) for time periods 1, 2 and 3, respectively. There was a significant decline in S. pneumoniae (P = 0.017) and increase in H. influenzae (P = 0.012) isolations and of H. influenzae that were beta-lactamase-producing (P = 0.04) among middle ear fluid isolates. Also there was a trend for an increased proportion of S. pneumoniae in 2001-2003 that were penicillin-susceptible (P = 0.17). CONCLUSIONS: In our experience, persistent AOM and AOMTF has decreased in frequency since the introduction of high dose amoxicillin therapy and pneumococcal conjugate vaccination. It appears that H. influenzae has become the predominant pathogen of persistent AOM and AOMTF since universal immunization with the pneumococcal conjugate vaccine. Fewer S. pneumoniae AOM isolates are penicillin-resistant and more H. influenzae are beta-lactamase-producing. PMID- 15361721 TI - Community-wide vaccination with the heptavalent pneumococcal conjugate significantly alters the microbiology of acute otitis media. AB - BACKGROUND: Community-wide use of conjugated heptavalent pneumococcal vaccine (PCV7) in children <2 years of age could affect the microbiology of acute otitis media (AOM) in vaccinees, particularly for penicillin-nonsusceptible Streptococcus pneumoniae (PNSP). SETTING: Since Summer 2000, 94% of young children cared for by this 7-clinician, pediatric practice in rural central Kentucky received 3 or 4 doses of PCV7 in the first 18 months of life. OBJECTIVE: To determine changes in microbiology of AOM before and after community-wide routine implementation of PCV7. METHODS: Among children 7-24 months old with severe or refractory AOM, we compared 336 AOM isolates from 1992-1998 with 83 AOM isolates from 2000-2003 in children who had received 3 or 4 doses of PCV7. RESULTS: Comparing each cohort (1992-1998 versus 2000-2003), the proportion of S. pneumoniae decreased from 48% to 31% (P = 0.009; relative risk, 0.754; 95% confidence interval, 0.628-0.906), and nontypable Haemophilus influenzae increased from 41% to 56% (P = 0.01; relative risk, 1.87; 95% confidence interval, 1.15-3.04; beta-lactamase-positive, 56% versus 64%, not significant). The proportions of intermediate PNSP and resistant PNSP, respectively, were 16% and 9% versus 13% and 6% pre- and post-PCV7, respectively. Vaccine and vaccine related serotypes, respectively, comprised 70% and 8% versus 36% and 32% of S. pneumoniae strains (P = 0.003). Post-PCV7, Gram-negative bacteria and beta lactamase-producing organisms accounted for two-thirds and one-half of all AOM isolates, respectively. DISCUSSION: The overall proportion of S. pneumoniae isolates and vaccine serotypes in AOM were significantly reduced by community wide use of PCV7 vaccine in our practice. The proportion of Gram-negative bacteria became 2-fold more frequent than S. pneumoniae in AOM in PCV7-vaccinated young children where PCV7 uptake was community-wide and supply was adequate. PMID- 15361722 TI - Comparison of five-day cefdinir treatment with ten-day low dose amoxicillin/clavulanate treatment for acute otitis media. AB - BACKGROUND: Short course beta-lactam antibiotic therapy for acute otitis media (AOM) should improve patient adherence, but it has not been evaluated since the heptavalent pneumococcal conjugate vaccine became routinely used in the United States. METHODS: In a prospective, investigator-blinded, multicenter study, 425 patients, age 6 months-6 years, with a clinical diagnosis of nonrefractory AOM were randomized to receive either 5 days of cefdinir therapy (14 mg/kg divided twice daily) or 10 days of amoxicillin/clavulanate therapy (45/6.4 mg/kg divided twice daily). Clinical response was assessed at end of therapy (2-4 days postantibiotic, respectively) and week 4 (study days 25-28). RESULTS: With no difference in demographics between treatment groups, overall the mean age (+/-SD) was 2.8 +/- 1.8 years, 65% had received conjugated pneumococcal vaccination and 48% had bilateral AOM. The satisfactory clinical response rate at end of therapy was comparable for cefdinir versus amoxicillin/clavulanate (88%, 170 of 194 versus 85%, 164 of 192; 95% CI -4.9, 9.3). Although this must be interpreted with caution, cefdinir showed an apparent trend for higher efficacy than amoxicillin/clavulanate (92%, 72 of 78 versus 77%, 55 of 71; P = 0.019) in a subsample of patients 6-24 months old who had received conjugated pneumococcal vaccination. The incidence of drug-related adverse events was less for cefdinir than for amoxicillin/clavulanate (24%, 50 of 211 versus 38%, 82 of 214; P = 0.0018) CONCLUSION: For children with nonrefractory AOM, based only on clinical endpoints, 5 days of therapy with cefdinir 14 mg/kg divided twice daily was comparable overall with 10 days of therapy with low dose amoxicillin/clavulanate 45/6.4 mg/kg divided twice daily. PMID- 15361723 TI - Modifying acute otitis media: implications of immunization with pneumococcal conjugate vaccine. PMID- 15361724 TI - Impact of haemophilus influenzae type b conjugate vaccine in South Africa and Argentina. AB - INTRODUCTION: Haemophilus influenzae type b (Hib) persists as a major cause of pediatric meningitis and pneumonia in developing countries in which Hib conjugate vaccines are not used. Demonstration of decreases in severe Hib disease after countries introduce Hib conjugate vaccine will help justify the resources necessary to purchase and provide the vaccine. Because surveillance for culture confirmed Hib meningitis is not available in many countries, alternative means to measure the impact of Hib conjugate vaccine would be useful. METHODS: Laboratory records from the years before and after introduction of the Hib conjugate vaccine were reviewed at 4 hospitals, 2 in Argentina and 2 in South Africa. Potential indicators of bacterial meningitis including cerebrospinal fluid (CSF) culture, white blood cell count, appearance, protein and glucose were recorded. RESULTS: After introduction of Hib conjugate vaccine, culture-confirmed Hib meningitis declined significantly at 3 of 4 hospitals (2 in Argentina and 1 in South Africa). In the same 3 hospitals, there was a significant decline after vaccine introduction in some of the following CSF indicators of bacterial meningitis: proportion of CSF specimens with white blood cell count > or = 100 x 10(6)/L, 500 x 10(6)/L and 1,000 x 10(6)/L; glucose <40 mg/dL; protein >100 mg/dL; and turbid appearance. CONCLUSIONS: Culture-confirmed Hib meningitis declined at 3 of the 4 hospitals after Hib vaccine introduction. Surrogate indicators of bacterial meningitis also declined and might be useful measures of Hib conjugate vaccine impact at hospitals where capacity to culture Hib is not available. PMID- 15361725 TI - Transmission of cytomegalovirus from mothers to preterm infants by breast milk. AB - OBJECTIVES: To assess the risk of transmission of cytomegalovirus (CMV) by breast milk from CMV-seropositive mothers to their breast-fed preterm infants and to evaluate their outcome. PATIENTS AND METHODS: The study population comprised breast-fed preterm infants with a birth weight of <1,500 g and gestational age of <35 weeks. Venous blood samples from the mothers and infants were tested for CMV IgG and IgM antibodies on the 5th and 30th day after birth. Breast milk was obtained for CMV DNA detection by polymerase chain reaction and viral culture on the 5th day and on the 3rd, 6th and 12th week. Urine samples of the babies were collected at the same time for CMV culture. Neurodevelopmental assessment was done at 6 months of age, corrected for preterm birth. RESULTS: Thirty-eight mothers and 42 infants (including 4 sets of twins) were enrolled in the study. A mother-infant pair was excluded because of inadequate breast milk collection. Thirty-six mothers (97.3%) were CMV-seropositive. CMV DNA of breast milk was detected in 35 seropositive mothers. Six infants of 5 mothers were infected (infected group) at a mean of 77 days after birth, and 34 infants of 31 mothers were not (noninfected group). In all the mothers of the infected group, CMV virus could be cultured from the milk whey. The average maternal CMV IgG on day 5 after delivery was higher in the infected than in the noninfected group. Sepsis-like symptoms and hyperbilirubinemia were more frequently noted in the infected infants than in the noninfected, but the difference was not statistically significant. Neurodevelopmental outcome did not significantly differ between the 2 groups. CONCLUSIONS: The risk of CMV infection in breast-fed premature infants was highest when the mothers shed viable virus in their breast milk. These mothers had high CMV IgG, which may help identify those mother-infant pairs at risk. Inactivation of the virus in milk by freezing may be a way of reducing the transmission of this virus via breast milk. PMID- 15361726 TI - Measuring antibody responses to a live attenuated influenza vaccine in children. AB - BACKGROUND: Hemagglutination inhibition (HAI) assay is the standard method for evaluating inactivated influenza vaccines, but no standard assay has been established for evaluating live attenuated influenza vaccines (LAIV). LAIV containing A/Beijing/262/95(H1N1) induced low serum HAI antibody responses to the antigenic variant, A/New Caledonia/20/99(H1N1) in a serologic study but provided protection against the A/New Caledonia-like viruses in a community study. Neutralization and HAI assays were compared by measuring H1N1 cross-reactive antibody responses to the LAIV in children. METHODS: Sera were collected from 50 children 1-8 years of age before vaccination and 4-6 weeks after each dose of the LAIV. Antibody titers to the 3 vaccine viruses were measured by the HAI assay, whereas antibody titers against the H1N1 vaccine virus (A/Beijing/262/95) and 2 H1N1 antigenic variants (A/Shenzhen/227/95 and A/New Caledonia/20/99) were measured by the HAI and neutralization assays. RESULTS: Initially seronegative participants were more likely to develop HAI seroconversion responses to the 3 vaccine viruses than the baseline seropositive participants (77% versus 14% for H1N1, 100% versus 20% for H3N2, 100% versus 19% for B, P < 0.01, Fisher's exact test). For the H1N1 cross-reactive antibody responses, seroconversion rates measured by the neutralization assay were significantly higher than those measured by the HAI assay (95% versus 78%, P = 0.0485 for A/Beijing/262/95; 75% versus 24%, P < 0.0001 for A/Shenzhen/227/95; 51% versus 5%, P < 0.0001 for A/New Caledonia/20/99). CONCLUSIONS: The neutralization assay was more sensitive than the HAI assay for measuring H1N1 antibody responses after vaccination of children with the LAIV and may provide a better correlate of clinical protection provided by the LAIV. PMID- 15361728 TI - Reducing illness transmission from disinfected recreational water venues: swimming, diarrhea and the emergence of a new public health concern. AB - Recreational water-related illnesses are associated with swimming in contaminated water venues. The transmission of diarrheal illness in disinfected settings is influenced by several factors which include: chlorine resistance in waterborne pathogens; poor facility maintenance of disinfectant levels; and lack of healthy swimming habits. Health care providers can help to disseminate healthy swimming messages to their patients and help to prevent and control this emerging public health concern. PMID- 15361727 TI - Two dosages of clarithromycin for five days, amoxicillin/clavulanate for five days or penicillin V for ten days in acute group A streptococcal tonsillopharyngitis. AB - BACKGROUND: Short course antimicrobial therapy is suggested for group A streptococcal tonsillopharyngitis. METHODS: The bacteriologic and clinical efficacies of clarithromycin [30 or 15 mg/kg/day twice daily (b.i.d.)] or amoxicillin/clavulanate (43.8/6.2 mg/kg/day b.i.d.) for 5 days or penicillin V (30 mg/kg/day 3 times a day) for 10 days were compared. In a randomized, open label, parallel group, multicenter study, 626 children (2-16 years old) with tonsillopharyngitis were enrolled; 537 were evaluable for efficacy. Follow-up evaluations were performed at 4-8 and 21-28 days after therapy. RESULTS: At enrollment, 26% of the Streptococcus pyogenes isolates were clarithromycin nonsusceptible. All regimens had an apparently similar clinical efficacy. The long term S. pyogenes eradication rates were 102 of 123 (83%) with amoxicillin/clavulanate and 88 of 114 (77%) with penicillin V. In the 30- and 15 mg/kg/day clarithromycin groups, eradication occurred in 71 of 86 (83%) and 59 of 80 (74%) of the clarithromycin-susceptible isolates (P = 0.33), and in 4 of 28 (14%) and 5 of 26 (19%) of the clarithromycin-resistant isolates, respectively (clarithromycin-susceptible versus -resistant, P < 0.0001). Both clarithromycin dosages were well-tolerated. CONCLUSIONS: In group A streptococcal tonsillopharyngitis, 5 days of clarithromycin or amoxicillin/clavulanate treatment had clinical efficacy comparable with that of 10 days of penicillin V treatment; however, amoxicillin/clavulanate and penicillin V were bacteriologically more effective than clarithromycin because of its failure to eradicate the clarithromycin-resistant S. pyogenes isolates. The 5-day clarithromycin regimens are not recommended for treatment of streptococcal tonsillopharyngitis in areas where in vitro resistance of group A streptococci to clarithromycin is common. PMID- 15361729 TI - Varicella infections and varicella vaccine in the 21st century. PMID- 15361730 TI - Complications of human immunodeficiency virus therapy. PMID- 15361731 TI - Association of regulatory issues with an echovirus 18 meningitis outbreak at a children's summer camp in Alaska. AB - We document an echovirus 18 meningitis outbreak occurring at a remote overnight children's camp in Alaska. The outbreak involved 26% of 113 camp residents, was associated with building overcrowding and occurred in a camp with a contaminated drinking water source. Lack of specific children's camp regulations and failure to implement and enforce existing regulations may have contributed to the outbreak. PMID- 15361732 TI - Successful treatment of a child with posttraumatic necrotizing fasciitis caused by Apophysomyces elegans: case report and review of literature. AB - Apophysomyces elegans is an uncommon human pathogen that causes deeply invasive infections in immunocompromised patients and cutaneous infection in immunocompetent patients. We report the development of severe deep soft tissue zygomycosis caused by A. elegans in an otherwise healthy child after trauma. She was successfully treated with surgical debridements and antifungal therapy with liposomal amphotericin B. A review of the literature indicates that zygomycosis caused by A. elegans is associated with traumatic inoculation. PMID- 15361733 TI - Sleeping sickness in brothers in london. AB - Brothers 9 and 14 years of age presented in London with fever and skin lesions after a safari in East Africa. Malaria films were negative, but trypanosomes were seen in blood films and chancre fluid. Sleeping sickness should be considered in children returning from East Africa. PMID- 15361734 TI - Pandoraea bacteremia in a cystic fibrosis patient with associated systemic illness. AB - Pandoraea is a recently classified genus primarily isolated from the sputum of cystic fibrosis patients, but its pathogenic potential is unknown. We describe a case of Pandoraea bacteremia in a 16-year-old cystic fibrosis patient associated with clinical disease, suggesting that this organism should be considered a true pathogen in susceptible patients. PMID- 15361735 TI - Hemophagocytic syndrome and Burkholderia cepacia splenic microabscesses in a child with chronic granulomatous disease. AB - Hemophagocytic syndrome, splenic microabscesses and pulmonary cavitary lesions were presented in a 17-month-old boy with prolonged fever, hepatosplenomegaly and a history of tuberculous lymphadenitis. Clinical course mimicked tuberculosis. Blood cultures were negative. Ultrasound-guided, percutaneous aspiration from splenic microabscesses grew Burkholderia cepacia. He was treated successfully with trimethoprim-sulfamethoxazole. This child with chronic granulomatous disease had an unusual clinical manifestation of B. cepacia infection. PMID- 15361736 TI - Tuberculous meningitis complicated with hydrocephalus and cerebral salt wasting syndrome in a three-year-old boy. AB - Cerebral salt wasting syndrome (CSWS) is a syndrome of hyponatremia and natriuresis described in patients with intracranial diseases. We describe a 3 year-old boy with tuberculous meningitis complicated by hydrocephalus and CSWS and emphasize the different clinical presentation and management of patients with CSWS and the syndrome of inappropriate secretion of antidiuretic hormone. PMID- 15361737 TI - Enhanced culture detection of Kingella kingae, a pathogen of increasing clinical importance in pediatrics. AB - Osteoarticular infection and occult bacteremia are the 2 invasive infectious pathologies most frequently associated in childhood with Kingella kingae. We report a series of 11 patients in whom osteomyelitis predominates over septic arthritis, which is the reverse of other series, probably as a consequence of inoculation of samples during surgery on agar media, used in combination with or as an alternative to inoculation into blood culture bottles. Although K. kingae infections usually follow a benign clinical course, we noted 2 patients with mild orthopedic sequelae. PMID- 15361738 TI - Osteonecrosis in children with severe acute respiratory syndrome. AB - Osteonecrosis is a debilitating bone disease affecting adults who have recovered from severe acute respiratory syndrome in Hong Kong and China, but there are no data on its prevalence in children. We report 5 children with magnetic resonance imaging evidence of osteonecrosis. In view of the high prevalence and asymptomatic presentation of osteonecrosis, we suggest magnetic resonance imaging screening for osteonecrosis in children with severe acute respiratory syndrome. PMID- 15361739 TI - Cerebrospinal fluid beta 2-microglobulin values in perinatally acquired cytomegalovirus infection. PMID- 15361740 TI - Pulmonary mycobacterium avium intracellulare infection in an immunocompetent child. PMID- 15361741 TI - Explanation of mathematical model. PMID- 15361743 TI - Disability reduction in elderly patients with schizophrenia. AB - This article reviews a frequently overlooked subject, the topic of schizophrenia in late life. By examining the available literature on schizophrenia in this particular population, we hope to provide clinicians with a better understanding of the distinguishing characteristics, course, and optimal treatments of this disease in elderly patients. The validity of the concept of symptom "burn out" is discussed and the cognitive changes seen in schizophrenia as patients age are examined. Similarities and differences between late-onset schizophrenia and early onset schizophrenia in aging patients are compared. The similarities and differences between schizophrenia and dementia in the elderly are also discussed. Finally, treatments for the illness, including both typical and atypical antipsychotic treatments, as well as nonpharmacological intervention strategies, along with their advantages and disadvantages, are reviewed. PMID- 15361744 TI - Comorbidity of anxiety and depressive disorders: issues in conceptualization, assessment, and treatment. AB - OBJECTIVES: The purpose of this review is to provide a clinically relevant analysis of issues concerning comorbidity among anxiety and depressive disorders. The co-occurrence of social anxiety disorder (SAD) and generalized anxiety disorder (GAD) with depressive disorders is highlighted as an illustration. Data on prevalence, rates of comorbidity, order of onset, course, and functional impairment associated with these disorders, in both the general population and clinical samples, are examined. The second half of the review focuses on discussion of practical issues concerning assessment and treatment of comorbid anxiety and depressive syndromes. CONCLUSIONS: Available evidence suggests that comorbidity among SAD, GAD, and the depressive disorders is substantial and pervasive. Co-occurrence of these syndromes is typically characterized by a chronic course with clinically significant impairment in social and occupational functioning. SAD and GAD precede the onset of major depression in a majority of cases and appear to be risk factors for developing major depression. Clinicians encountering patients with primary complaints of anxiety or depression should carefully assess for the presence of comorbid symptoms and syndromes. Treatment outcome research suggests that pharmacotherapy and psychosocial therapy (cognitive-behavior therapy in particular) both represent viable first-line treatment alternatives. However, with increasing severity of depression, pharmacotherapy is indicated as a primary intervention. The authors recommend increased efforts in screening and detection, more clinical trials that include patients with comorbid syndromes and symptoms, and continued research on the integration of pharmacological and psychotherapeutic treatments. PMID- 15361745 TI - Comparability of telephone and face-to-face interviews in assessing patients with posttraumatic stress disorder. AB - INTRODUCTION: Structured clinical interviews are very important in the area of mental health research and services. Prior research comparing the reliability and validity of face-to-face and phone interviews has found high levels of agreement. This project compared the results of face-to-face and phone interviews for two widely used measures: The Clinician-Administered PTSD Scale (CAPS) for assessing posttraumatic stress disorder diagnostic status and symptom severity and the Hamilton Rating Scale for Depression (Ham-D) to determine the severity of major depressive disorder. METHOD: Subjects were 34 veterans recruited from applicants to the PTSD Assessment and Intervention program at the Cincinnati VA Medical Center. Order of interview (in-person or phone) was determined using random assignment within a counterbalanced framework. After attaining satisfactory levels of interrater reliability, four clinicians independently and blindly evaluated the subjects. RESULTS: Pearson correlation coefficients between face-to face and phone interviews revealed high consistency (CAPS r = 0.745, HAM-D r = 0.748). The level of agreement between the two methods was 82% for the CAPS and 85% for the HAM-D. Diagnostic thresholds for the CAPS and HAM-D, after adjusting for the interview order and time elapsed between interviews, did not differ between the two groups (p = 0.31 for the CAPS and p = 0.96 for the HAM-D). High levels of agreement were achieved between the two methods (kappa = 0.75 for the CAPS using a cutoff of 65 and 0.70 for the HAM-D). The high sensitivity, specificity, and predictive values support the reliability of the phone-interview method. CONCLUSION: Phone interviews are a reliable method of interviewing for use in assessing patients for posttraumatic stress disorder and major depressive disorder. PMID- 15361746 TI - Cognitive-behavioral assessment of body image disturbances. AB - The construct of body image is multidimensional in nature and multidisciplinary in scope. The purpose of this review is to help clinicians navigate the basic theoretical and practical considerations associated with body image assessment. Common pitfalls reflect methodological problems and conceptual confusion surrounding the subjectivity, heterogeneity, lability, and complexity of the body image construct. By highlighting these core concepts, we provide a framework for conducting a comprehensive cognitive-behavioral assessment to inform case formulation and treatment planning. The strength and utility of a cognitive behavioral therapy (CBT) model lies in its broad applicability and ability to identify the proximal factors that precipitate and maintain body image concerns, such as distorted patterns of thinking and maladaptive behavior (e.g., body checking, mirror gazing, avoidance, social comparison, reassurance seeking). Although the evidence base is still evolving, the basic tenets of a CBT approach appear to transcend disciplines and specialties. The article concludes by providing an example of self-monitoring, which is a cornerstone of CBT assessment and treatment, and referencing the steps of an established CBT manualized treatment program. PMID- 15361747 TI - Tianeptine: a facilitator of the reuptake of serotonin and norepinephrine as an antidepressant? PMID- 15361748 TI - Cognitive-behavioral therapy for schizophrenia. PMID- 15361749 TI - An ethical dilemma: when the family wants the withdrawal of care. AB - The ethical principles of beneficence, nonmaleficence, and respect for the dignity and autonomy of the patient provide the moral-ethical foundation for the doctor-patient relationship. However, inability to obtain informed consent in the absence of advance directives as well as conflicting family sentiments can give rise to ethical dilemmas. We present a case of a 59-year-old man who survived a near fatal suicide attempt by shooting himself in the face and whose family communicated their desire for the patient to have medical care withheld. The ethical issues that emerged from this suicide attempt, including the family's perspective on the patient's right to die, are discussed in the context of a consultation-liaison psychiatrist's recommendations to a multi-specialty treatment team. PMID- 15361750 TI - Antihypertensive therapy and new onset diabetes. PMID- 15361751 TI - Facts and fiction of poor compliance as a cause of inadequate blood pressure control: a systematic review. AB - OBJECTIVE: (i) To obtain an overview of measured compliance with antihypertensive medication; (ii) to explore sources of variation in measured compliance percentages between studies; and (iii) to investigate whether studies can establish a relationship between compliance and achieved blood pressure. STUDY SELECTION: MEDLINE, PubMed and EMBASE databases were searched for studies, published between 1985 and 2003, in which patient compliance with antihypertensive medication was measured with electronic monitors. DATA EXTRACTION: A standard form was used to extract relevant data from the included articles. Two investigators extracted all data independently. RESULTS: Thirty studies were identified. The studies included were highly varied in their methods used to quantify compliance. Taking compliance and/or correct dosing were used in 20 studies. Mean taking compliance for a once-daily regimen was 94.0 +/- 4.4% and 88.2 +/- 6.5% for a twice-daily regimen. Mean correct dosing was 85.0 +/- 8.7% for a once-daily regimen and 75.3 +/- 6.5% for a twice-daily regimen. Proportions of patients with taking compliance or correct dosing /= 2 days. Subjects wore an ambulatory BP monitor after the experiments. Repeated measure analysis of variance (ANOVA) tested if BP and heart rate differed over time and between experimental conditions. Multivariate regression tested factors related to the BP response. RESULTS: For 9 h after all experiments, average awake systolic blood pressure (SBP) increased and diastolic blood pressure (DBP) decreased compared with baseline (P < 0.001). Average awake SBP increased up to 6.9 mmHg less (P < 0.001) and DBP decreased 2.6 mmHg more (P < 0.05) after exercise versus control. For 5 h, PEH was greater after MOD; but over the course of 9 h, LITE was as effective as MOD in eliciting PEH. Baseline BP was the primary factor explaining the BP response (beta = -0.434 to -0.718, r = 0.096-0.295). CONCLUSIONS: LITE and MOD evoked PEH throughout the daytime hours. Lower intensity dynamic exercise such as walking, contributes to BP control in men with hypertension. PMID- 15361759 TI - Systemic hemodynamics in non-anesthetized L-NAME- and DOCA-salt-treated mice. AB - OBJECTIVE: Long-term cardiac output measurements in non-anesthetized mice are now possible. We used this technology to study two different hypertensive models in mice. DESIGN: We combined telemetric blood pressure and heart rate recordings with Doppler flow probe cardiac output measurements in mice during treatment with Nomega-nitro-L-arginine methyl ester (L-NAME) and deoxycorticosterone acetate (DOCA)-salt. METHOD: The mice received a flowprobe around the ascending aorta and, 10-18 days later, blood pressure telemetry. After recovery, baseline values were recorded and the mice were given L-NAME (5 mg/10 ml tap water), L-NAME followed by valsartan (50 mg/kg per day per gavage), or DOCA-salt (50 mg DOCA pellet, 0.9% saline to drink, uninephrectomy). Mean arterial pressure, heart rate, stroke volume and cardiac output were recorded daily and total peripheral resistance was calculated. RESULTS: L-NAME resulted in an abrupt increase in mean arterial pressure caused solely by an increase in total peripheral resistance. Cardiac output was decreased. Valsartan treatment decreased blood pressure and total peripheral resistance, while cardiac output was restored to normotensive values. DOCA-salt required 3 days before hypertension developed. Contrary to the volume expansion, increased cardiac output, autoregulation hypothesis, the blood pressure increase was only associated with increased total peripheral resistance, while cardiac output was not changed. CONCLUSION: Both L-NAME and DOCA-salt increased blood pressure by increasing total peripheral resistance. Comprehensive hemodynamics can be done in non-anesthetized, free-moving mice. The methods provide new perspectives for studying mouse models in the long-term. PMID- 15361760 TI - Aldosterone synthase gene (CYP11B2) C-344T polymorphism, plasma aldosterone, renin activity and blood pressure in a multi-ethnic population. AB - BACKGROUND: The aldosterone synthase gene (CYP1B2) locus is a candidate region involved in the development of hypertension. OBJECTIVE: To study the relationship between the C-344T CYP1B2 polymorphism, plasma aldosterone, renin activity and blood pressure in a multi-ethnic population. DESIGN: Population-based, cross sectional study of 1313 middle-aged men and women (456 white, 441 of African origin and 416 South Asian). Anthropometry, blood pressure, biochemistry, questionnaire data and timed urine collections were taken with standardized techniques. All were genotyped for the C-344T CYP11B2 polymorphism. RESULTS: The frequency of the C allele was significantly lower in people of African origin (0.21) than in white (0.46) and South Asian (0.43) (P < 0.001). After adjustment for age, sex and ethnicity the TT genotype was associated with 14% higher plasma aldosterone levels, 3.7 mmHg higher systolic and 2.1 mmHg higher diastolic blood pressure than CC (P for linear trend < 0.05). No significant interactions with age, sex, ethnicity, body mass index (BMI) and fractional excretion of sodium were found in the associations between genotype and both blood pressure and aldosterone levels. In a sub-sample of participants in which plasma renin activity was measured (n = 457), a significant excess of T alleles was found in those with a raised (>/= 750) aldosterone-to-renin ratio (ARR). CONCLUSION: In this multi-ethnic population, the C-344T CYP1B2 polymorphism is associated with blood pressure, plasma aldosterone levels and ARR. Although significant differences in allele frequencies were found between groups, ethnicity does not explain the results. PMID- 15361761 TI - Two medium-chain acyl-coenzyme A synthetase genes, SAH and MACS1, are associated with plasma high-density lipoprotein cholesterol levels, but they are not associated with essential hypertension. AB - BACKGROUND AND OBJECTIVES: SAH has been proposed as a candidate gene for essential hypertension (EH) because elevated expression of SAH was observed in the kidneys of spontaneously hypertensive rats. Recently, a homology search of SAH in the human genome revealed the presence of the SAH gene family, which includes SAH, MACS1, MACS2, and MACS3. SAH and MACS1 are located within a 150-kb region on human chromosome 16p13.11. SAH and MACS1 are thought to function as acyl-coenzyme A synthetases, which are involved in fatty acid metabolism. In the present study, we analyzed six single nucleotide polymorphisms (SNPs) in the SAH and MACS1 genes in a Japanese population, and examined whether these SNPs contribute to EH and multiple risk factors. METHODS AND RESULTS: We performed association studies of six SNPs in 287 EH patients and 259 normotensive subjects. Multiple logistic linear regression analysis revealed that the allele frequencies of these six SNPs in SAH and MACS1 genes were not significantly different between EH patients and normotensives. SNP in exon 8 of the A/G polymorphism of the MACS1 gene and the G/T SNP in intron 3 of the SAH gene were associated with plasma levels of plasma high-density lipoprotein cholesterol. CONCLUSIONS: SNPs in the MACS1 and SAH genes contribute to plasma levels of high-density lipoprotein cholesterol. PMID- 15361762 TI - Endothelin-B-receptor-selective antagonist inhibits endothelin-1 induced potentiation on the vasoconstriction to noradrenaline and angiotensin II. AB - OBJECTIVE: Endothelin-A-receptor-antagonists inhibit angiotensin II- and noradrenaline-induced vasoconstriction. Whether functional constrictive endothelin-B-receptors play a role in the endothelin-1-mediated potentiation of vasoconstriction to angiotensin II and noradrenaline is thus far unknown. METHODS: We studied the effects of noradrenaline and angiotensin II (10 mol/l) in the presence of exogenous endothelin-1 (10 mol/l) with and without selective endothelin-B-receptor-blockade by BQ-788 (10 mol/l) and dual receptor blockade with BQ-788 and the endothelin-A-selective antagonist BQ-123 (10 mol/l) in 14 healthy male volunteers (aged 20-28). Studies were performed in the human skin microcirculation under in vivo conditions using laser-Doppler flowmetry and double injection technique. The area under the time-response curve of all doses was calculated. RESULTS: Endothelin-1 potentiated the effects of angiotensin II and noradrenaline (-944 +/- 139 perfusion units (PU), P < 0.01; -926 +/- 117 PU, P < 0.05, respectively). In the presence of BQ-788, the potentiating effect of endothelin-1 was significantly blunted (-624 +/- 132 PU, P < 0.01; -549 +/- 136 PU, P < 0.01, respectively). In the presence of BQ-123 and BQ-788 the vasoconstriction was fully inhibited (431 +/- 108 PU, P < 0.001 and 421 +/- 86 PU, P < 0.001, respectively). CONCLUSIONS: These data suggest that functional vasoconstrictive endothelin-B receptors on vascular smooth muscle cells may contribute to the potentiating effects of high local concentrations of endothelin 1 on the vasoconstriction to noradrenaline and angiotensin II in human microcirculation. PMID- 15361763 TI - beta-Adrenoceptor-mediated, nitric-oxide-dependent vasodilatation is abnormal in early hypertension: restoration by L-arginine. AB - BACKGROUND: It is unknown whether beta-adrenoceptor-mediated vasodilatation is altered in early hypertension and whether it can be modulated by L-arginine. METHODS AND DESIGN: We measured changes in forearm blood flow by plethysmography in response to acetylcholine (9 and 37 microg/min), sodium nitroprusside (200 and 800 ng/min) and the beta-receptor agonist, isoproterenol (50 and 200 ng/min) in 12 patients with essential hypertension (group EH) and in healthy volunteers with (group PFH; n = 14) and without (group NFH; n = 14) a family history of essential hypertension, before and during concomitant infusion of L-arginine (10 micromol/min). In five individuals from each group, infusion of acetylcholine and isoproterenol was repeated during the concurrent blockade of nitric oxide synthesis by N-monomethyl-L-arginine (L-NMMA; 4 micromol/min). RESULTS: The response to acetylcholine was reduced in groups EH and PFH compared with group NFH (both P < 0.05), whereas the vasodilator effects of isoproterenol and sodium nitroprusside were similar in all three groups. Acetylcholine- and isoproterenol induced vasodilatation did not change during infusion of the nitric oxide precursor, L-arginine, in group NFH, but were significantly enhanced by L arginine in groups PFH and EH [forearm blood flow before and after isoproterenol 200 ng/min: group PFH 11.8 +/- 1.02 and 13.3 +/- 1.08 ml/min, respectively (P < 0.05); group EH: 11.3 +/- 1.57 and 14.9 +/- 1.91 ml/min, respectively (P < 0.01)]. Co-infusion of L-NMMA blunted the response to acetylcholine and isoproterenol in group NFH (P < 0.05), but did not significantly modify vasodilatation in groups PFH and EH. CONCLUSIONS: Although beta-adrenergic vasodilatation seemed to be unaltered in early hypertension, L-arginine enhanced the response to isoproterenol, whereas concomitant inhibition of nitric oxide synthase by L-NMMA had no significant effect. These findings suggest that the nitric oxide component of isoproterenol-mediated vasodilatation is impaired in early hypertension and possibly compensated by increased beta-adrenoceptor responsiveness of smooth muscle cells. In this setting, supplementation of the nitric oxide precursor, L-arginine, enhances the vasodilator response to beta adrenergic stimulation. PMID- 15361764 TI - L-S-nitrosothiols: endothelium-derived hyperpolarizing factors in porcine coronary arteries? AB - OBJECTIVE: Bradykinin-induced, endothelium-derived hyperpolarizing factor (EDHF) mediated responses depend on Ca-dependent K-channels (KCa) of small (SKCa) and intermediate (IKCa) conductance, inwardly rectifying K (KIR) channels and/or Na-K ATPase. Here we investigated in porcine coronary arteries (PCAs) whether S nitrosothiols can act as EDHF. METHODS: Preconstricted PCAs were exposed to bradykinin, the NO donor S-nitroso-N-penicillamine (SNAP), or the S-nitrosothiols L-S-nitrosocysteine (L-SNC), D-SNC and L-S-nitrosoglutathione (L-SNG), with or without KCl, the NO scavenger hydroxocobalamin, the S-nitrosothiol-depleting agent p-hydroxymercurobenzoic acid (PHMBA) and/or inhibitors of NO synthase (L NAME), guanylyl cyclase (ODQ), SKCa channels (apamin), KCa channels of large conductance (BKCa) (iberiotoxin), IKCa + BKCa channels (charybdotoxin), KIR channels (BaCl2) or Na-K-ATPase (ouabain). RESULTS: All agonists concentration dependently relaxed PCAs. L-NAME, charybdotoxin + apamin, KCl, and ouabain shifted the bradykinin concentration-response curve (CRC) approximately 10-fold to the right. BaCl2 did not exert additional effects on top of ouabain. Full blockade of bradykinin was obtained when combining L-NAME with charybdotoxin + apamin, KCl or ouabain + BaCl2. PHMBA reduced the maximum effect of bradykinin. Iberiotoxin + apamin, alone or on top of L-NAME, did not affect bradykinin, SNAP or L-SNC. ODQ and hydroxocobalamin shifted the SNAP, L-SNC, D-SNC, and L-SNG CRCs approximately 10-fold to the right, and, in combination, fully blocked SNAP induced effects. Charybdotoxin + apamin shifted the L-SNC and L-SNG CRCs, but not the D-SNC or SNAP CRCs, approximately 5-fold to the right. KCl and ouabain (but not BaCl2) shifted the SNAP, L-SNC and L-SNG CRCs 5-10 fold to the right. CONCLUSIONS: L-S-nitrosothiols activate SKCa + IKCa channels in a stereoselective manner, whereas NO activates Na-K-ATPase. Since S-nitrosothiols decompose to NO, stored L-S-nitrosothiols may mediate bradykinin-induced, EDHF-dependent relaxation. PMID- 15361765 TI - Increased pulse wave velocity is not associated with elevated augmentation index in patients with diabetes. AB - OBJECTIVE: Increased arterial stiffness is a risk factor for cardiovascular disease and is a feature associated with diabetes. Pulse wave velocity (PWV) is an accepted index of arterial stiffness and augmentation index (AI) derived from radial applanation tonometry has been advocated as a measurement of arterial stiffness. This study compares the relationship between PWV and AI in people with and without diabetes. DESIGN AND METHODS: A total of 66 people with diabetes and 66 age-matched non-diabetic controls were studied. Central aortic pressure waves were generated using applanation tonometry over the radial artery and used to calculate AI. Carotid-femoral PWV (PWVcf) was measured simultaneously. RESULTS: Relative to controls, diabetes was associated with increased pulse pressure (PP) and PWVcf (P < 0.01). In contrast, AI did not differ between groups even after adjustment for heart rate. This observation remained consistent irrespective of diabetes type, arterial site, and the presence or absence of antihypertensive therapy. Multiple regression analysis revealed diabetes to be a significant determinant of PWVcf, but not AI. CONCLUSIONS: PP and PWVcf are increased in people with diabetes, but this is not associated with increased AI. These findings conclusively demonstrate that AI is not a reliable measure of arterial stiffness in people with diabetes. PMID- 15361766 TI - Plasma adrenomedullin and carotid atherosclerosis in atherothrombotic ischemic stroke. AB - OBJECTIVE: Adrenomedullin is known to exert anti-atherosclerotic actions by inhibiting proliferation and migration of smooth muscle cells in vitro. Here we examine the relationship between the plasma concentration of adrenomedullin and ultrasonographic characteristics of carotid arteries both in ischemic stroke and in the absence of cerebrovascular disease. METHODS: We studied 61 patients with atherothrombotic ischemic stroke in the chronic phase and 50 patients without any cerebrovascular disease. Intima-media thickness and vascular lumen diameters were evaluated by carotid ultrasonography. Plasma mature-adrenomedullin was determined by radioimmunoassay. RESULTS: Plasma mature-adrenomedullin in the patients with atherothrombotic ischemic stroke in the chronic phase (2.01 +/- 0.58 fmol/ml) was significantly higher than that in the patients without any cerebrovascular disease (1.24 +/- 0.18 fmol/ml, P < 0.001). With multiple regression analysis, plasma mature-adrenomedullin was found to be predicted by: stroke status (atherothrombotic ischemic stroke versus no cerebrovascular disease), diabetes status (yes/no), left ventricular ejection fraction, internal carotid artery intima-media thickness, and common carotid artery pressure strain elastic modulus (R = 0.79; F5,105 = 85.39, P < 0.0001). CONCLUSION: Plasma mature-adrenomedullin showed significantly positive associations with carotid atherosclerosis and atherothrombotic ischemic stroke, independent of systolic blood pressure. PMID- 15361767 TI - Effects of adrenomedullin on cell proliferation in rat adventitia induced by aldosterone. AB - OBJECTIVE: Aldosterone is involved in cardiovascular diseases such as hypertension and heart failure by inducing sodium retention and vascular remodeling, which is characterized by fibroblast proliferation and migration in adventitia. It is well known that aldosterone stimulates vascular smooth muscle cells and fibroblasts to produce and secrete adrenomedullin (ADM), a multiple functional peptide with an important cytoprotective effect against cardiovascular damage. We examined the effect of aldosterone on ADM production and secretion and its mRNA expression in rat aortic adventitia to study the paracrine/autocrine interaction between endogenous ADM and aldosterone. METHODS: ADM produced and secreted from adventitia stimulated by aldosterone in the absence or presence of spironolactone, RU486 or spironolactone together with RU486 were detected by radioimmunoassay, proliferation in adventitia cells was evaluated by the level of [H]-thymine incorporation, and preproADM gene expression was measured by semi quantitative reverse transcriptase polymerase chain reaction. RESULTS: Adventitial ADM secretion and mRNA expression stimulated by aldosterone were concentration-dependent as was the inhibitive effect of ADM on aldosterone induced proliferation. The induction of aldosterone in ADM secretion was mediated by mineralocorticoid receptor. Antagonists of specific receptors of calcitonin gene-related peptide (CGRP) receptor type 1 and ADM both potentiated the proliferation effect induced by aldosterone; and thiorphan, an inhibitor of the enzyme for ADM degradation, inhibited the adventitial [H]-thymine incorporation induced by aldosterone. ADM inhibited the activity of extracellular signal related kinase (ERK) stimulated by aldosterone. CONCLUSION: Aldosterone stimulates adventitia to produce and secrete ADM, which in turn, antagonizes the aldosterone-induced proliferation in adventitia. PMID- 15361768 TI - Rho kinase and PAI-1 in Bartter's/Gitelman's syndromes: relationship to angiotensin II signaling. AB - OBJECTIVE: Angiotensin II (Ang II)-mediated activation of Rho kinase (ROK) is involved in the pathophysiology of hypertension and cardiovascular remodeling. ROK also controls plasminogen activator inhibitor-1 (PAI-1) which promotes vascular fibrosis contributing to atherogenesis. Bartter's and Gitelman's syndromes (BS/GS) are useful models to investigate abnormalities of vascular tone regulation, due to their reduced short- and long-term signaling pathways of Ang II. This study evaluated, using BS/GS as a model, ROK and PAI-1 gene and protein expression and the effect of Ang II co-incubation on ROK and PAI-1 gene and protein expression. DESIGN, METHODS AND RESULTS: We measured ROK and PAI-1 gene and protein expression [reverse transcription-polymerase chain reaction (RT-PCR) and Western blot] in mononuclear cells (PBM) from one BS and eight GS patients. The effect of Ang II on ROK and PAI-1 gene and protein expression was also evaluated and compared with 10 controls. ROK gene and protein expression was reduced in BS/GS [0.47 +/- 0.11 densitometric units (d.u.) versus 0.70 +/- 0.04 d.u., P = 0.0038 and 0.39 +/- 0.07 d.u. versus 0.55 +/- 0.07 d.u., P = 0.0026, respectively]. The basal level of PAI-1 gene and protein expression did not differ (0.40 +/- 0.03 d.u. versus 0.39 +/- 0.02 d.u. and 0.81 +/- 0.02 d.u. versus 0.83 +/- 0.02 d.u., respectively). Ang II increased ROK and PAI-1 gene and protein expression only in controls: from 0.70 +/- 0.04 to 0.90 +/- 0.06 d.u., P = 0.007 (ROK mRNA); from 0.55 +/- 0.07 to 0.86 +/- 0.07 d.u., P = 0.0005 (ROK protein); from 0.40 +/- 0.02 to 0.63 +/- 0.03 d.u., P = 0.001 (PAI-1 mRNA); and from 0.83 +/- 0.02 to 1.34 +/- 0.16 d.u., P = 0.0023 (PAI-1 protein). CONCLUSIONS: This study confirms BS/GS as a human model to investigate interrelated systems involved in the pathophysiology of hypertension and throws more light on the cellular mechanisms of BS/GS reduced Ang II short- and long term signaling pathways. PMID- 15361769 TI - Evidence against a major role for angiotensin converting enzyme-related carboxypeptidase (ACE2) in angiotensin peptide metabolism in the human coronary circulation. AB - OBJECTIVE: To investigate the role of angiotensin-converting enzyme-related carboxypeptidase (ACE2) in angiotensin peptide metabolism in the human coronary circulation. METHODS: Angiotensin I and angiotensin II, and their respective carboxypeptidase metabolites, angiotensin-(1-9) and angiotensin-(1-7), were measured in arterial and coronary sinus blood of heart failure subjects receiving angiotensin-converting enzyme (ACE) inhibitor therapy and in normal subjects not receiving ACE inhibitor therapy. In addition, angiotensin I, angiotensin II and angiotensin-(1-7) were measured in arterial and coronary sinus blood of subjects with coronary artery disease before, and at 2, 5 and 10 min after, intravenous administration of ACE inhibitor. RESULTS: In comparison with normal subjects, heart failure subjects receiving ACE inhibitor therapy had a greater than 40-fold increase in angiotensin I levels, but angiotensin-(1-9) levels were low (1-2 fmol/ml), and similar to those of normal subjects. Moreover, angiotensin-(1-7) levels increased in parallel with angiotensin I levels and the angiotensin-(1 7)/angiotensin II ratio increased by 7.5-fold in coronary sinus blood. Intravenous administration of ACE inhibitor to subjects with coronary artery disease rapidly decreased angiotensin II levels by 54-58% and increased angiotensin I levels by 2.4- to 2.8-fold, but did not alter angiotensin-(1-7) levels or net angiotensin-(1-7) production across the myocardial vascular bed. CONCLUSIONS: The failure of angiotensin-(1-9) levels to increase in response to increased angiotensin I levels indicated little role for ACE2 in angiotensin I metabolism. Additionally, the levels of angiotensin-(1-7) were more linked to those of angiotensin I than angiotensin II, consistent with its formation by endopeptidase-mediated metabolism of angiotensin I, rather than by ACE2-mediated metabolism of angiotensin II. PMID- 15361770 TI - Blockade of the renin-angiotensin system decreases adipocyte size with improvement in insulin sensitivity. AB - OBJECTIVE: Based on results of in vitro studies, it has been hypothesized that blockade of the renin-angiotensin system (RAS) promotes the recruitment and differentiation of pre-adipocytes and that increased formation of small insulin sensitive adipocytes counteracts ectopic deposition of lipids, thereby improving insulin sensitivity. We investigated the effect of RAS blockade on insulin sensitivity, adipocyte size, and intramuscular lipid content in fructose-fed rats (FFR) as a model of insulin-resistant hypertension. DESIGN AND METHODS: Six-week old male Sprague-Dawley rats were divided into two groups: those fed a standard chow (control) and those fed a fructose-rich chow for 6 weeks. FFR were treated with a vehicle or with 1 mg/kg per day of temocapril, an angiotensin-converting enzyme inhibitor, or 0.1 mg/kg per day of olmesartan, an angiotensin II type 1 receptor blocker, for the last 2 weeks. Insulin sensitivity (M value: mg/kg per min) was estimated by the euglycemic hyperinsulinemic glucose clamp method. Sizes of adipocytes derived from epididymal fat and triglyceride content in the soleus muscle were determined. RESULTS: FFR had lower M value, higher blood pressure, larger adipocyte size, higher ratio of epididymal fat pads over body weight (%fat pads), and higher intramuscular triglyceride than did the control rats. Both temocapril and olmesartan significantly improved the M value and decreased blood pressure and adipocyte size without change in %fat pads in FFR. Adipocyte size was negatively correlated with the M value. Treatment for 2 weeks decreased, but not significantly, intramuscular triglyceride. CONCLUSIONS: RAS blockade decreases adipocyte size without change in epididymal %fat pads accompanied by improvement in insulin sensitivity. PMID- 15361771 TI - Effect of fish diets and weight loss on serum leptin concentration in overweight, treated-hypertensive subjects. AB - BACKGROUND: Leptin, a circulating hormone secreted from adipocytes, is an index of adiposity and is reduced by caloric restriction and weight loss. A recent population study suggested that dietary-derived omega3 fatty acids lower leptin levels independent of body fat. OBJECTIVE: To examine whether dietary fish enhanced the effects of weight loss on serum leptin levels, in 69 overweight, treated hypertensive men and women. METHODS: Participants were randomized to a daily fish meal, a weight-reduction regimen, the two regimens combined or a control group for 16 weeks. RESULTS: A total of 63 individuals completed the study. Weight fell 5.6 +/- 0.8 kg with energy-restriction. Blood pressure (BP) reductions in the combined fish-weight loss group were twice that seen with either intervention alone. At baseline, in all groups combined, serum leptin levels correlated with serum insulin (r = 0.307, P = 0.014), but not with body weight. The greatest change in serum leptin occurred in the fish-weight loss group (control, 0.60 +/- 0.76 ng/ml; fish, 1.20 +/- 0.79 ng/ml; weight loss, 1.40 +/- 1.05 ng/ml; fish-weight loss, -5.08 +/- 1.64 ng/ml). In the fish-weight loss group, the change in serum leptin was predicted by changes in serum insulin (r = 0.488, P = 0.038), 24-h BP (systolic BP (SBP): r = 0.435, P = 0.060; diastolic BP (DBP): r = 0.563, P = 0.018) and 24-h heart rate (0.584, P = 0.028). Using general linear models, there was a significant fish x weight-loss interaction (P = 0.008) on post-intervention serum leptin after adjustment for baseline levels, independent of post-intervention insulin. CONCLUSION: A daily fish meal as part of a weight-reducing regimen was more effective than either measure alone at reducing leptin levels. Reductions in leptin may be related to the substantial fall in BP seen with the fish-weight loss program. PMID- 15361772 TI - Metabolic syndrome and target organ damage in untreated essential hypertensives. AB - BACKGROUND: The prevalence and the relationship between metabolic syndrome, and target organ damage (TOD) in essential hypertensive patients has not been fully explored to date. OBJECTIVE: To investigate the association between metabolic syndrome, as defined by the ATP III report, and cardiac and extracardiac TOD, as defined by the 2003 ESH-ESC guidelines for management of hypertension, in a large population of never-treated essential hypertensives. METHODS: A total of 447 grade 1 and 2 hypertensive patients (mean age 46 +/- 12 years) who were attending a hypertension hospital outpatient clinic for the first time underwent the following procedures: (i) physical examination and repeated clinic blood pressure measurements; (ii) routine examinations; (iii) 24-h urine collection for microalbuminuria; (iv) 24-h ambulatory blood pressure monitoring; (v) echocardiography; and (vi) carotid ultrasonography. Metabolic syndrome was defined as involving at least three of the following alterations: increased waist circumference, increased triglycerides, decreased high-density lipoprotein cholesterol, increased blood pressure, or high fasting glucose. Left ventricular hypertrophy (LVH) was defined according to two different criteria: (i) 125 g/m in men and 110 g/m in women; (ii) 51 g/h in men and 47 g/h in women. RESULTS: The 135 patients with metabolic syndrome (group I) were similar for age, sex distribution, known duration of hypertension and average 24-h, daytime and night time ambulatory blood pressure to the 312 patients without it (group II). The prevalence of altered left ventricular patterns (LVH and left ventricular concentric remodelling) was significantly higher in group I (criterion a = 30%, criterion b = 42%) than in group II (criterion a = 23%, criterion b = 30%, P < 0.05 and P < 0.01, respectively). A greater urinary albumin excretion (17 +/- 35 versus 11 +/- 23 mg/24 h, P = 0.04) was also found in group I compared to group II. There were no significant differences between the two groups in the prevalence of carotid intima-media thickening and plaques. CONCLUSIONS: These results from a representative sample of untreated middle-aged hypertensives show that: (i) the metabolic syndrome is highly prevalent in this setting and (ii) despite similar ambulatory blood pressure values, patients with metabolic syndrome have a more pronounced cardiac and extracardiac involvement than those without it. PMID- 15361773 TI - Attenuation of haemodynamic, metabolic and energy expenditure responses to isoproterenol in patients with hypertension. AB - OBJECTIVE: Overweight and heightened sympathetic activity are more common in hypertensive than normotensive subjects. beta-adrenoceptor down-regulation has been described in hypertension. We tested the hypothesis that chronic sympathetic overactivity impairs beta-adrenergic-mediated thermogenesis and thereby favours gain of weight in hypertension. PARTICIPANTS: The study included 13 hypertensive subjects aged 35.3 +/- 7.9 years and 25 normotensive subjects of control of similar age. METHODS: To measure beta-adrenergically mediated haemodynamic, metabolic and thermogenic responsiveness, increasing doses of isoproterenol diluted in 2.5 ml saline were injected as intravenous boluses (0.1, 0.25, 0.5, 1.0 and 2.0 microg/m). On a separate day, isoproterenol was infused continuously intravenously in increasing doses (10, 20 and 40 ng/kg per min), each dose for 30 min. RESULTS: The sitting heart rate and body mass were greater in hypertensives (P = 0.000, and P = 0.005, respectively). The heart rate responses to 1 and 2 microg/m isoproterenol bolus (P = 0.01 and P = 0.03, respectively) were reduced in hypertensives. The energy expenditure (P = 0.002) and oxygen consumption (P = 0.0004) increase with 40 ng/kg per min isoproterenol infusion, and glucose and phosphate responses at both 20 (P = 0.01 and P = 0.05) and 40 (P = 0.001 and P = 0.02) ng/kg per min isoproterenol infusion were attenuated in hypertensives. The baseline heart rate negatively correlated with heart rate (P = 0.015) response to isoproterenol bolus and blood pressure (P = 0.02) response to isoproterenol infusion. The urinary noradrenaline negatively correlated with heart rate response to isoproterenol bolus (P = 0.001), and with systolic blood pressure (P = 0.02) and energy expenditure responsiveness to isoproterenol infusion (P = 0.04). Furthermore, plasma noradrenaline negatively correlated with heart rate responsiveness to isoproterenol bolus (P = 0.004). CONCLUSIONS: These results show a generalized decrease of beta-adrenergic responsiveness in stage 1 hypertension and support the concept that sympathetic overactivity, via down regulation of beta-adrenoceptor-mediated thermogenic responses, may facilitate the development of obesity in hypertension. PMID- 15361774 TI - Relations between insulin sensitivity, fitness and autonomic cardiac regulation in healthy, young men. AB - OBJECTIVES: We hypothesized that insulin sensitivity and vagal cardiac control are independently related in young men after adjustment for fitness and other confounding variables. DESIGN: Male volunteers aged 21-24 years with high (borderline hypertensive; n = 20) and low-normal (normotensive; n = 21) screening blood pressure (BP) were studied cross-sectionally. METHODS: Mean R-R interval (RR) and heart rate variability (HRV) were computed from 30-min ECGs, and baroreflex sensitivity (BRS) and latency (phase shift) from 15-min beat-to-beat finger blood pressure (BP) and heart rate recordings. Insulin-adjusted glucose disposal rate (GDR/I) was measured with a 90-min hyperinsulinaemic glucose clamp and fitness by peak oxygen uptake (VO2peak) during a treadmill test. RESULTS: HRV, baroreflex function, GDR/I, and VO2peak did not differ between the groups. GDR/I correlated positively with time and frequency domain HRV, including high frequency power (HF) (r = 0.40, P = 0.01) and root-mean squared successive differences (RMSSD) (r = 0.43, P = 0.005), but not BRS or phase shift. GDR/I correlated with VO2peak (r = 0.70, P < 0.0001) and was explained (R = 0.56) by VO2peak (beta = 0.57, P < 0.0001) and RR (beta = 0.29, P = 0.03), independently of HRV and measures of obesity. Conversely, RR (beta = 0.55, P = 0.0004) and HRV, including HF (beta = 0.44, P = 0.006) and RMSSD (beta = 0.46, P = 0.004) were explained by GDR/I, independently of VO2peak. CONCLUSIONS: Insulin sensitivity and autonomic cardiac control are related independently of physical fitness in young men. PMID- 15361775 TI - The systemic haemodynamic and cerebral autoregulatory effects of bendrofluazide in the subacute post-stroke period. AB - INTRODUCTION: Little data exist on the efficacy in terms of blood pressure reduction or outcome measures for the various antihypertensive agents in patients post-stroke. In this study the effects of bendrofluazide on blood pressure levels and variability, dynamic cerebral autoregulation and cardiac baroreceptor sensitivity were assessed in the sub-acute stroke period. METHODS: A total of 36 hypertensive ischaemic stroke patients were randomized to oral bendrofluazide 2.5 mg daily or matching placebo starting 10 days post-ictus and continued for 28 days. A total of 12 hypertensive controls were similarly randomized in a double blind, crossover study, each limb being of 28 days duration. Cerebral blood flow velocity, non-invasive beat-to-beat blood pressure levels, electrocardiograms (ECGs) and transcutaneous carbon dioxide levels were measured before and at the end of each treatment period in stroke and control subjects. RESULTS: Casual blood pressure levels were not significantly reduced in stroke patients with bendrofluazide, but in the placebo group levels increased by 13 +/- 13/6 +/- 7 mmHg (P < 0.001) at study termination. In the control group, casual systolic blood pressure fell during active treatment by 12 +/- 16 mmHg (P < 0.03) compared with placebo. Dynamic cerebral autoregulation, beat-to-beat blood pressure variability and cardiac baroreceptor sensitivity did not change with bendrofluazide or placebo in either the stroke patients or controls. CONCLUSION: Bendrofluazide does not appear to be an effective hypotensive agent in the sub acute post-stroke period but may limit the blood pressure rise seen during stroke recovery. Bendrofluazide however does not adversely influence dynamic cerebral autoregulation, blood pressure variability or cardiac baroreceptor sensitivity in either strokes or control subjects. PMID- 15361776 TI - Superoxide dismutase, catalase and glutathione peroxidase in the spontaneously hypertensive rat kidney: effect of antioxidant-rich diet. AB - BACKGROUND AND OBJECTIVE: Earlier studies have shown increased production of reactive oxygen species (ROS) and upregulation of ROS-generating enzyme, nicotinamide adenine dinucleotide (phosphate) oxidase, in the kidney of spontaneously hypertensive rats (SHR). This study aimed to examine the activities and protein abundance of the main antioxidant enzymes [i.e. superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX)] in the kidney of SHR fed a regular or an antioxidant-rich diet. METHODS: Pregnant SHR and their offspring were fed either a regular diet or an antioxidant-rich diet (alpha-tocopherol, ascorbic acid, zinc and selenium) and observed for 6 months. Wistar-Kyoto (WKY) rats fed a regular or antioxidant-fortified diet served as controls. RESULTS: The untreated SHR showed severe hypertension and significant increases in plasma hydrogen peroxide and renal tissue nitrotyrosine abundance, indicating the presence of oxidative/nitrosative stress. Despite oxidative stress, Cu Zn SOD, CAT and GPX activities were unchanged in the cortex and medulla of untreated SHR. Immunodetectable Mn SOD was reduced in the medulla and elevated in the cortex, whereas, Cu Zn SOD protein was unchanged in the cortex and reduced in the medulla. By contrast, CAT protein abundance was increased in both cortex and medulla while GPX protein was elevated in the cortex and unchanged in the medulla. Comparison of protein abundance and activities of the antioxidant enzymes revealed significant discordance in the untreated SHR. Lifelong antioxidant therapy diminished the severity of hypertension, improved oxidative stress and ameliorated or reversed abnormalities of antioxidant enzyme expressions and activities. By contrast, antioxidant therapy had no effect on the measured parameters in the WKY rat controls. CONCLUSIONS: Oxidative stress in SHR was associated with a lack of coordinate upregulation of the antioxidant enzymes and discordance between their protein abundance and enzymatic activity. These findings suggest an impaired antioxidant defense system and the presence of functionally abnormal enzymes in the SHR kidney. Lifelong antioxidant therapy improved expression, activity and activity-to-mass relationship of the measured enzymes. The latter suggests oxidative and nitrosative modification of these molecules in the SHR kidney. PMID- 15361777 TI - Effects of telmisartan 80 mg and valsartan 160 mg on ambulatory blood pressure in patients with essential hypertension. PMID- 15361779 TI - The current epidemic of primary aldosteronism: causes and consequences. PMID- 15361781 TI - The current epidemic of primary aldosteronism: causes and consequences. PMID- 15361783 TI - The current epidemic of primary aldosteronism: causes and consequences. PMID- 15361784 TI - Will autoantibodies help to determine severity and progression of atherosclerosis? PMID- 15361785 TI - Mechanism of lipoprotein retention by the extracellular matrix. AB - PURPOSE OF REVIEW: Considerable evidence suggests that the subendothelial retention of atherogenic lipoproteins is a key early step in atherogenesis. In humans and experimental animals, elevated levels of plasma lipoproteins are associated with increased atherosclerosis, and lipoproteins with higher affinity for arterial proteoglycans are more atherogenic. Here we discuss the molecular mechanisms underlying lipoprotein retention in the arterial wall and how this interaction can be modulated. RECENT FINDINGS: Functional proteoglycan binding sites in lipoproteins containing apolipoprotein B have been identified and shown to have atherogenic potential in vivo. In addition to apolipoprotein B, novel bridging molecules, those that can interact with both proteoglycans and lipoproteins, have been identified that mediate the retention of atherogenic particles in the vessel wall. The interaction between lipoproteins and proteoglycans can be enhanced by the modification of lipoproteins in the circulation and in the arterial wall, by alterations in the subendothelium, and by changes in proteoglycan synthesis that result in a more atherogenic profile. The retention of atherogenic lipoproteins is a potential target for therapies to reverse atherosclerosis, and in-vitro studies have identified compounds that decrease the affinity of proteoglycans for lipoproteins. SUMMARY: Considerable progress has been made in understanding the association between lipoproteins and cardiovascular disease. This review highlights the importance of the interaction between lipoproteins and the arterial matrix. PMID- 15361786 TI - Functional role for toll-like receptors in atherosclerosis and arterial remodeling. AB - PURPOSE OF REVIEW: Activation of inflammatory cascades is causally related to the development of atherosclerotic disease. Toll-like receptors are innate immune receptors that recognize pathogen-associated molecular patterns. In this review the pathways by which toll-like receptors might play a role in the development and progression of atherosclerosis will be discussed according to recent literature. RECENT FINDINGS: Toll-like receptors are expressed in atherosclerotic tissue. Next to pathogens, endogenous toll-like receptor ligands have been linked with the development of arterial occlusive disease. In mouse models of hyperlipidemia, a potential role for the toll-like receptor pathway has been suggested in hypercholesterolemia-induced atherosclerosis. Recent in-vitro studies revealed a mechanism by which toll-like receptor ligation results in a strong inhibition of cholesterol efflux from macrophages. In addition, oxidized lipoproteins interact with toll-like receptors. Furthermore, activation of the apoptotic cascade, which is important during atherogenesis, enhances the toll like receptor pathway resulting in upregulation of proinflammatory cytokines. Human epidemiologic studies have linked TLR4 polymorphism with atherosclerosis. However, data on the association between atherosclerosis progression and TLR4 polymorphisms are conflicting. Next to plaque growth, arterial remodeling is an important determinant of luminal narrowing in atherosclerosis. Recently, a possible role for TLR4 signaling in arterial remodeling has been revealed in mouse models. SUMMARY: A clarification of the molecule [corrected] mechanisms by which the toll-like receptor signaling cascade influences atherosclerosis might [corrected] lead to novel strategies to intervene in the development of this life threatening disease. PMID- 15361787 TI - Caveolin-1 and caveolae in atherosclerosis: differential roles in fatty streak formation and neointimal hyperplasia. AB - PURPOSE OF REVIEW: Caveolae are 50-100 nm cell surface plasma membrane invaginations observed in terminally differentiated cells. They are characterized by the presence of the protein marker caveolin-1. Caveolae and caveolin-1 are present in almost every cell type that has been implicated in the development of an atheroma. These include endothelial cells, macrophages, and smooth muscle cells. Caveolae and caveolin-1 are involved in regulating several signal transduction pathways and processes that play an important role in atherosclerosis. RECENT FINDINGS: Several recent studies using genetically engineered mice (Cav-1 (-/-) null animals) have now clearly demonstrated a role for caveolin-1 and caveolae in the development of atherosclerosis. In fact, they suggest a rather complex one, either proatherogenic or antiatherogenic, depending on the cell type examined. For example, in endothelial cells, caveolin-1 and caveolae may play a proatherogenic role by promoting the transcytosis of LDL cholesterol particles from the blood to the sub-endothelial space. In contrast, in smooth muscle cells, the ability of caveolin-1 to negatively regulate cell proliferation (neointimal hyperplasia) may have an antiatherogenic effect. SUMMARY: Caveolin-1 and caveolae play an important role in several steps involved in the initiation of an atheroma. Development of new drugs that regulate caveolin 1 expression may be important in the prevention or treatment of atherosclerotic vascular disease. PMID- 15361788 TI - Acid sphingomyelinase in macrophage apoptosis. AB - PURPOSE OF REVIEW: This article presents an overview of the recent progress in understanding metabolic and functional interrelationships of biologically active sphingolipids related to the sphingomyelin signal transduction pathway in relation to the regulation of apoptosis in macrophages. RECENT FINDINGS: Ceramide generation is an essential, early step in apoptosis in numerous systems. There are several mechanisms for ceramide generation, including activation of plasma membrane, lysosomal, nuclear, and mitochondrial sphingomyelinases, and induction of de-novo synthesis of ceramide. Some of the proapoptotic actions of ceramide are to facilitate assembly of death receptor complexes in the plasma membrane, to prevent the activation of protein kinase B/Akt, and to promote the activation of caspase 3. Failure of macrophages in developing atherosclerotic plaques to undergo apoptosis is a possible contributor to plaque expansion. At low concentrations, oxidized LDL has been shown to prevent apoptosis induced by growth factor withdrawal in cultured bone marrow-derived macrophages, in part by inhibiting sphingomyelinase and preventing generation of ceramide. At high concentrations, however, oxidized LDL can induce apoptosis or necrosis of macrophages. SUMMARY: Sphingolipid signal transduction pathways play an important role in the regulation of growth and survival pathways in macrophages. These are directly relevant to the pathogenesis of a variety of chronic inflammatory disorders, including atherosclerosis. PMID- 15361789 TI - Lysosomal acid lipase and atherosclerosis. AB - PURPOSE OF REVIEW: Atherosclerosis remains the leading cause of death in the developed countries. In addition to lipid-lowering drugs - statins, dietary control, and exercise, new approaches are needed for the treatment and prevention of atherosclerosis. This review will focus on the role(s) of lysosomal acid lipase and its use as an enzyme therapy to reduce atherosclerotic lesions in a mouse model and to examine the molecular basis supporting this novel strategy and its mechanism of effect. RECENT FINDINGS: Administration of human lysosomal acid lipase via tail vein into mice with atherosclerosis eliminates early aortic and coronary ostial lesions and reduces lesional size in advanced disease. The reduction of advanced lesional area is related to decreases in foamy macrophages, collagen positive areas, and necrotic areas. Compared with sham-treated mice, the human lysosomal acid lipase-treated mice also have reduced levels of plasma cholesteryl esters, and reduced levels of hepatic cholesterol and triglycerides. SUMMARY: These studies indicate that administrated lysosomal acid lipase affects the atherogenesis by at least two mechanisms: (1) direct targeting of lesional macrophages with resultant decreases in cholesteryl esters and triglyceride in the lysosomes of macrophages in the lesions; (2) systemic effects that mediate the liver to reduce the hepatic cholesteryl ester and triglyceride release, possibly leading to reduced production of VLDL and LDL. PMID- 15361790 TI - Blocking endothelial adhesion molecules: a potential therapeutic strategy to combat atherogenesis. AB - PURPOSE OF REVIEW: This review provides a concise update of the involvement of endothelial adhesion molecules in atherogenesis, an overview of current advances in the development of adhesion molecule blocking agents, as well as an insight into the potential of these molecules in cardiovascular therapy. RECENT FINDINGS: As endothelial adhesion molecules are deemed to play an important role in the development and progression of atherosclerotic lesions, they are interesting targets for therapeutic intervention in this process. In particular, P-selectin and vascular cell adhesion molecule 1 are widely considered to hold promise in this regard. Current research efforts centre on the design of agents that directly block the interaction of the receptor with its ligand (e.g. soluble P selectin glycoprotein ligand 1, blocking antibodies, EWVD-based peptides) or that interfere with their synthesis (e.g. antisense oligonucleotides) or their regulatory control by nuclear factor kappa B or peroxisome proliferator-activated receptor gamma. Furthermore, adhesion molecules have been exploited as a target for the specific delivery of drug carriers (e.g. biodegradable particles with entrapped dexamethasone) or therapeutic compounds (e.g. dexamethasone) to the plaque. All approaches have been shown to be effective in blocking adhesion molecule function in in-vitro studies and in-vivo models for inflammation or atherosclerosis. SUMMARY: Although the field has achieved considerable progress in recent years, leading to the development of a number of interesting leads, final proof of their efficacy in cardiovascular therapy is eagerly awaited. PMID- 15361791 TI - Mechanisms of leukocyte recruitment to atherosclerotic lesions: future prospects. AB - PURPOSE OF REVIEW: Leukocyte invasion in the arterial wall is critical in the development of atherosclerotic lesions. This review describes recent advances in the understanding of leukocyte recruitment in atherogenesis and in the development of vulnerable plaque. It also discusses limitations in the current knowledge of this process and how these limitations may be addressed. RECENT FINDINGS: The adhesive function of platelets has recently been highlighted as an important recruitment mechanism in atherosclerosis. For example, targeted deficiency of P-selectin in platelets reduces atherosclerosis in mice. Platelets also increase monocyte recruitment in atherosclerosis by secreting chemokines such as platelet factor 4 (CXCL4) or RANTES (CCL5), which trigger monocyte arrest in atherosclerotic arteries. A causal role for RANTES in atherosclerosis was shown by a protective effect of the blockage of RANTES receptors in apolipoprotein E-deficient mice. A similar effect was also demonstrated for the fractalkine receptor CX3CR1. Moreover, the classic chemoattractant LTB4 plays important roles in atherosclerosis, inasmuch as the absence of the principal LTB4 receptor (BLT1) reduces early atherosclerosis in mice. Novel data have also shown that many types of cells in lesions express 5-lipoxygenase, which indicates a rich source of leukotrienes in plaque. SUMMARY: Recent data provide evidence for the involvement of several adhesive and signalling mechanisms in leukocyte recruitment in atherosclerosis. However, the specific mechanisms that are responsible for the accumulation of proatherogenic leukocytes in lesions are unclear. Detailed study of certain subclasses of leukocytes in the recruitment process will be important in future studies in this field. PMID- 15361792 TI - Control of smooth muscle cell proliferation in vascular disease. AB - PURPOSE OF REVIEW: Smooth muscle cell proliferation has previously been regarded as a central feature in vascular disease. The role of this process has recently been substantially re-evaluated, and we have reconsidered the functional importance of smooth muscle cell proliferation, the origin of proliferating smooth muscle cells in lesions, and the mechanisms whereby smooth muscle cell proliferation is controlled. In this review, we summarize recent progress in the understanding of smooth muscle cell proliferation, with a particular focus on how interactions between the extracellular matrix, smooth muscle cells, and mitogens control critical steps in this process. RECENT FINDINGS: Irrespective of the origin of smooth muscle cells in vascular lesions, fundamental interactions between the extracellular matrix and cell surface integrins are necessary in order to initiate a proliferative response in a quiescent smooth muscle cell, in a similar manner to any non-malignant cell. These interactions trigger intracellular signaling and cell cycle entry, which facilitate cell cycle progression and proliferation by mitogens. In addition, extracellular matrix interactions may also control the availability and activity of growth factors such as heparin-binding mitogens, which can be sequestered by heparan sulfate containing extracellular matrix components and regulate smooth muscle cell proliferation. SUMMARY: New insights into mechanisms whereby the extracellular matrix takes part in the control of smooth muscle cell proliferation suggest a number of putative targets for future therapies that can be applied to increase plaque stability, prevent the clinical consequences of atherosclerosis and improve outcomes after interventional procedures and organ transplantation. PMID- 15361793 TI - Mast cells in vulnerable coronary plaques: potential mechanisms linking mast cell activation to plaque erosion and rupture. AB - PURPOSE OF REVIEW: A novel link between inflammation and acute coronary syndromes is emerging, in that infiltrating inflammatory cells may convert a clinically silent coronary plaque into a dangerous and potentially lethal plaque. The majority of acute atherothrombotic events now relate to erosion or rupture of such unstable plaques. Here we summarize the molecular mechanisms by which activated mast cells may contribute to plaque erosion or rupture. RECENT FINDINGS: In-vitro experiments have revealed a multitude of paracrine effects exerted by activated mast cells. By secreting heparin proteoglycans and chymase, activated mast cells efficiently inhibit the proliferation of smooth muscle cells in vitro, and reduce their ability to produce collagen by a transforming growth factor beta-dependent and -independent mechanism. Mast cell chymase and tryptase are capable of activating matrix metalloproteinases types 1 and 3, causing degradation of the extracellular matrix component, collagen, necessary for the stability of the plaque. Activated mast cells also secrete matrix metalloproteinases types 1 and 9 themselves. Furthermore, chymase induces SMC apoptosis by degrading fibronectin, a pericellular matrix component necessary for SMC adhesion and survival, with the subsequent disruption of focal adhesions and loss of outside-in survival signaling. By secreting chymase and tumour necrosis factor alpha, activated mast cells also induce endothelial cell apoptosis. SUMMARY: Locally activated mast cells may participate in the weakening of atherosclerotic plaques by secreting heparin proteoglycans, chymase, and cytokines, which affect the growth, function and death of arterial endothelial cells and smooth muscle cells, thereby predisposing to plaque erosion or rupture. PMID- 15361794 TI - The accumulation of specific types of proteoglycans in eroded plaques: a role in coronary thrombosis in the absence of rupture. AB - PURPOSE OF REVIEW: Although fibrous cap rupture is the primary cause of coronary thrombosis, plaque erosion is responsible for 30%-40% of acute thrombotic events. The interface of the eroded surface involves a denuded endothelium allowing direct contact of the platelet/fibrin thrombus with the underlying lesion. This review discusses the putative role of extracellular matrix molecules, in particular proteoglycans/hyaluronan, in the development of acute coronary thrombosis associated with erosion. RECENT FINDINGS: The plaque/thrombus interface in erosion presents a unique surface since it consists of predominantly SMCs and proteoglycans with minimal or no inflammation. The lack of significant inflammation raises the possibility that erosion represents chronic wounding rather than true atherogenesis. The abundance of proteoglycan and hyaluronan matrix suggests their potential role in the development of thrombosis. Matrix changes may contribute to endothelial loss, the magnitude of the thrombotic event, or both. Versican facilitates platelet adhesion at low shear and cooperates with collagen to promote platelet aggregation. Further, versican may, in part, regulate water content and in turn support coagulation because water dependent functionality of anticoagulation molecules. Finally, experimental models of plaque erosion are currently being developed guided by the premise that the loss of surface endothelium together with other procoagulant factors may underlie the development of platelet-rich thrombi. SUMMARY: The loss of endothelium and exposure of a potentially procoagulant versican-hyaluronan matrix may be largely responsible for plaque erosion. The development of relevant animal models should allow further insight into the pathophysiology of coronary thrombosis in the absence of rupture. PMID- 15361795 TI - Periodontal infections and atherosclerosis: mere associations? AB - PURPOSE OF REVIEW: Several lines of evidence from the last few decades suggest that periodontitis is an important risk factor for cardiovascular diseases. In this review we discuss the recent findings on the systemic effects of periodontitis, which may contribute to the pathogenesis of atherosclerosis, with a special emphasis on lipoproteins. RECENT FINDINGS: In addition to the epidemiological studies exploring the direct or indirect relationship between clinical periodontitis and cardiovascular diseases, studies utilizing serology, animal models, cell cultures, and biochemistry of lipoproteins have been published. Local infection in the periodontal pockets triggers a systemic inflammatory response releasing inflammatory mediators and awakens a strong immune response against periodontal pathogens. Elevated systemic antibody levels especially to Porphyromonas gingivalis are associated with an increased risk for atherosclerosis. Periodontitis is also accompanied by proatherogenic changes in both low and high density lipoproteins, which lead to enhanced cholesteryl ester uptake by and reduced cholesterol efflux from macrophages. Vesicles and lipopolysaccharide isolated from P. gingivalis activate macrophages to convert into foam cells. Moreover, animal studies have demonstrated that infection by P. gingivalis enhances progression of atherosclerosis. SUMMARY: Recent studies have clarified the mechanisms by which periodontitis may contribute to the development of atherosclerosis. Serological, animal, and cell culture studies provide evidence that infection by P. gingivalis may promote atherosclerosis. The influence of periodontitis on lipoprotein metabolism has emerged as a new, important factor. Recent studies provide experimental proof that periodontitis may predispose to atherosclerosis. PMID- 15361796 TI - Estrogen in cardiovascular disease. AB - PURPOSE OF REVIEW: The controversy surrounding hormone replacement therapy has induced fear in patients and left many researchers with the impression that estrogen produces negative effects on cardiovascular function. The aim of this review is to summarize recent findings illustrating that estrogen also has positive effects even if estrogen replacement therapy is not a cure-all. RECENT FINDINGS: Studies have unveiled new aspects of estrogen action in the cardiovascular system; however, clinical trials have not demonstrated a protective effect of the most widely used modalities of hormone replacement therapy against cardiovascular disease. New information has emerged showing that estrogen has both beneficial and detrimental effects. Further mechanistic studies and use of well defined forms of estrogens and selective estrogen receptor modulators will continue to provide novel mechanistic information that will likely lead to the development of new avenues for therapeutic interventions. SUMMARY: Estrogens, like other steroid hormones, are potent actors in the cardiovascular system. Since half the population have high levels of estrogen most of their lives it is plain that estrogen has a variety of beneficial physiologic functions. Clinical studies, however, have demonstrated that a specific formulation of a combination of potent estrogens and metabolites is not a magic bullet, but induces both positive and negative impacts on different organ systems. More research into the mechanistic actions of estrogens in specific pathways in individual cell types is necessary to determine appropriate therapeutic interventions to replace the loss of positive effects of estrogens while minimizing the negative effects in postmenopausal women. PMID- 15361797 TI - Bibliography. Current world literature. Atherosclerosis: cell biology and lipoproteins. PMID- 15361798 TI - Nutrition and metabolism. PMID- 15361799 TI - Genetics and molecular biology: the ABC of cholesterol efflux and high-density lipoprotein formation. PMID- 15361800 TI - Lipid metabolism. PMID- 15361801 TI - Hyperlipidaemia and cardiovascular disease: ALLHAT, low-fat diet and apolipoprotein measurements: are there emerging issues? PMID- 15361802 TI - Atherosclerosis: cell biology and lipoproteins: C-reactive protein is a biomarker of atherosclerosis and mediator of plaque formation. PMID- 15361803 TI - Therapy and clinical trials. PMID- 15361806 TI - Dosage and administration issues of antipsychotic agents (and adjunctive medicines) in the acute stabilization of psychoses. AB - There are several options and approaches available to practicing psychiatrists for the pharmacologic management of acute psychotic episodes that present in emergency room settings. This article focuses on the first hours and days of such episodes, the goals of treatment, and comparison of the "old favorites," such as combination haloperidol and lorazepam (in liquid, tablet or injectable formulations), with the newer second-generation antipsychotic agents, such as olanzapine, ziprasidone, risperidone, quetiapine, or aripiprazole, which are available in different formulations. The speed of titration to an effective target dose for these newer agents are addressed as well. PMID- 15361807 TI - Rethinking the treatment paradigm for bipolar depression: the importance of long term management. AB - The need for long-term management of bipolar disorder is evident. Bipolar patients spend more time depressed than manic; however, few agents used for maintenance therapy of bipolar disorder have demonstrated good efficacy in delaying relapse into depression. This article provides a comprehensive review of open-label and randomized, controlled studies examining prophylactic efficacy in bipolar disorder, especially bipolar depression. Lithium, considered the gold standard for bipolar disorder maintenance therapy may be more effective in delaying manic relapse than in delaying depressive relapse. Evidence for the efficacy of divalproex and carbamazepine in delaying depressive relapse is yet to be fully elucidated. Lamotrigine has demonstrated efficacy in delaying time to depressive relapse. Unpublished studies show olanzapine's efficacy in preventing manic recurrence, while its efficacy in preventing depressive recurrence is yet to be proven. As patients with bipolar disorder are prone to experiencing depressive episodes, more attention needs to be focused on preventing depressive relapse. To date, three agents--lithium, lamotrigine, and olanzapine--have been shown to have prophylactic benefits in treating this highly recurrent disorder. PMID- 15361808 TI - Combination treatments for schizophrenia. AB - Combination treatments, especially combinations of antipsychotics, are used frequently for schizophrenia, despite a paucity of evidence regarding their safety and efficacy. Because the literature basis is weak and expert recommendations are largely lacking, providers should be vigilant in documenting improved outcomes for patients thought to benefit from combination treatments. Target symptoms that have been studied include psychosis, cognitive deficits, and negative symptoms. The strongest evidence is for augmentation of clozapine with another antipsychotic or with electroconvulsive therapy for persistent positive symptoms. Combination treatments for cognitive deficits and negative symptoms are being actively investigated, but current evidence is insufficient to recommend available agents for these components of schizophrenia. It is important that appropriate monotherapies be given adequate trials before resorting to combination therapies. PMID- 15361809 TI - Effective dosing and dose equivalency of second-generation antipsychotic medications. AB - Dosing patterns with second-generation antipsychotic medications (SGAs) are dynamic, with some SGAs surpassing current recommendations while others are declining in dose since their initial regulatory guidelines. Pertinent recent studies and available pharmacoepidemiologic reports, information, and expert consenses are reviewed herein to illuminate current thinking on the topic of dosing with SGAs. There is a need for fixed-dose studies of each SGA. Additionally, dosing should be a primary consideration when designing and subsequently interpreting comparative studies between SGAs. PMID- 15361811 TI - The neurobiology of pleasure, reward processes, addiction and their health implications. AB - Modern science begins to understand pleasure as a potential component of salutogenesis. Thereby, pleasure is described as a state or feeling of happiness and satisfaction resulting from an experience that one enjoys. We examine the neurobiological factors underlying reward processes and pleasure phenomena. Further, health implications related to pleasurable activities are analyzed. With regard to possible negative effects of pleasure, we focus on addiction and motivational toxicity. Pleasure can serve cognition, productivity and health, but simultaneously promotes addiction and other negative behaviors, i.e., motivational toxicity. It is a complex neurobiological phenomenon, relying on reward circuitry or limbic activity. These processes involve dopaminergic signaling. Moreover, endorphin and endogenous morphinergic mechanisms may play a role. Natural rewarding activities are necessary for survival and appetitive motivation, usually governing beneficial biological behaviors like eating, sex and reproduction. Social contacts can further facilitate the positive effects exerted by pleasurable experiences. However, artificial stimulants can be detrimental, since flexibility and normal control of behavior are deteriorated. Additionally, addictive drugs are capable of directly acting on reward pathways. Thus, the concrete outcome of pleasant experiences may be a question of dose. Moderate pleasurable experiences are able to enhance biological flexibility and health. Hence, pleasure can be a resistance resource or may serve salutogenesis. Natural rewards are mediated by sensory organ stimulation, thereby exhibiting a potential association with complementary medical approaches. Trust and belief can be part of a self-healing potential connected with rewarding stimuli. Further, the placebo response physiologically resembles pleasure phenomena, since both involve brain's reward circuitry stimulation and subjective feelings of well being. Pleasurable activities can stimulate personal growth and may help to induce healthy behavioral changes, including stress management. However, more research is needed to better understand the nature, neurobiology and maybe dangerous aspects of pleasure. Also, a possible involvement of endogenous morphinergic signaling has to be studied further. PMID- 15361812 TI - Direct measurement of free radicals in the brain cortex and the blood serum after nociceptive stimulation in rats. AB - The concentrations of ROS were measured in samples of the sensorimotor brain cortex and in the rat blood. We measured the following parameters: The six lines spectra, nitroxide radical, free hydroxyl radical and singleton oxygen. Their concentration was measured under physiological conditions, after the nociceptive stimulation and after the application of melatonin, both in normal and stimulated animals. In the brain cortex only the singleton oxygen decreased after the nociceptive stimulation, whereas the nitroxide radicals and six lines spectra increased. The free hydroxyl radicals did not change significantly. In the blood serum the six lines spectra and nitroxide radical increased, the concentration of the free hydroxyl radicals did not change. Melatonin increased both the hydroxyl and nitroxide radicals. There was a non-significant decrease in the six lines spectra. The estimation of ROS can be used as a tool for detecting metabolic changes and the consequences of different environmental influences, in our case the influence of nociception and melatonin. PMID- 15361813 TI - Isoproterenol-stimulated melatonin production by perifused rat pineal glands: age and time-related effects. AB - OBJECTIVES: The purpose of this study was to investigate and compare the effects of the beta-adrenergic agonist, isoproterenol, on induced increase in melatonin production in the pineal gland of young and old rats, at different circadian stages. MATERIALS AND METHODS: We report here the effects of 10(-6) M isoproterenol-stimulated melatonin production by perifused pineal glands obtained from young (55 day old) and old (21 month old) male Wistar rats acclimatised to light:dark cycles regimen of 12:12 for 3 weeks. Pineal glands were collected at different circadian stages: 3, 7, 11, 15, 19 and 23 hours after light onset (HALO), and perifused for 510 min. RESULTS: The basal levels of melatonin production in the young rats were approximately twice greater than those of the old rats. Isoproterenol stimulated melatonin production in both young and old rat pineal glands, whatever the circadian stage. The intensity of the response to 10( 6) M isoproterenol infusion was greater in young than in old rat pineal glands (P<0.001), with a trend towards an increase during the light phase, at 7 HALO, in both young and old rat pineal glands, although this trend towards increased melatonin response did not reach statistical significance (P>0.05). CONCLUSIONS: These results show that isoproterenol is able to stimulate both young and old rat pineal glands whatever the circadian stage. The magnitude of isoproterenol stimulation is greater in young than in old glands. Our results also suggest that the pineal gland response to isoproterenol is not dependent on circadian stage, at least, under our experimental conditions. PMID- 15361814 TI - Cardiovascular risk factors in middle-aged women with subclinical hypothyroidism. AB - OBJECTIVES: Overt hypothyroidism (OH) is associated with premature atherosclerosis and coronary heart disease (CHD). Recently, C-reactive protein (CRP) and total homocysteine (tHct) emerged as additional independent cardiovascular risk factors. Subclinical hypothyroidism (SH), affecting as many as 15% of middle-aged women is not known to be associated with risk for CHD. DESIGN AND MEASUREMENTS: We measured CRP and tHct levels as well as conventional cardiovascular risk markers in 44 middle-aged women with newly diagnose SH. Results were compared with those obtained in 10 patients with OH and 19 euthyroid controls. RESULTS: In SH, tHct and CRP levels were not as augmented as compared to controls. Their mean systolic and diastolic blood pressure values were increased vs. controls (p<0.04;p<0.01, respectively). Mean values of total cholesterol (TC), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), triglycerides, TC/HDL-C and LDL-C/HDL-C were not different in patients with SH compared to controls. Individual analysis revealed that the percentage of patients with SH having hypertension, hypertriglyceridemia, hypercholesterolemia, elevated TC/HDL-C and LDL-C/HDL-C ratios were higher than the percentage in controls. CRP positively correlated with BMI(r=0.29,p<0.02), and tHct positively correlated with age (r=0.24, p<0.05). CONCLUSIONS: Our findings suggest that subclinical hypothyroidism in middle-aged women is associated with hypertension and dyslipidemia. CRP and tHct do not appear to contribute to the increased risk for CHD in these patients. PMID- 15361815 TI - Immune and stress responses in C57BL/6 and C3H/HeN mouse strains following photoperiod manipulation. AB - OBJECTIVES: The purpose of this study was to determine whether two different strains of mice, that are reproductively unresponsive to photoperiod, adjust immune function in response to photoperiod. SETTING AND DESIGN: Adult male C3H/HeN (C3H) and C57BL/6 (C57) mice were each placed into either a long (LD 16:8) or short (LD 8:16) photoperiod for 10 weeks (n = 15/ group). METHODS: Blood was collected for flow cytometry and radioimmunoassay analysis of leukocyte numbers and corticosterone concentrations, respectively. In addition, all mice were sensitized to, and challenged with the antigen, 2,4-dinitro-1-fluorobenzene. Pinnae measurements were obtained for 7 days following challenge to determine the magnitude of the inflammatory response. RESULTS: Photoperiod did not affect leukocyte cell numbers in either C3H or C57 mice. C3H mice displayed higher neutrophil numbers than C57 mice (p< 0.0001), whereas C57 mice displayed higher lymphocyte numbers than C3H mice (p< 0.01). C3H mice housed in LD 16:8 had higher corticosterone concentrations than those housed in LD 8:16 (p< 0.005) and C57 mice housed in either photoperiod (p< 0.05). Photoperiod did not affect the inflammatory response, though C57 mice displayed an overall higher magnitude of response than C3H mice (p<0.05). MAIN FINDINGS: Our results suggest that photoperiod does not alter immune function in C3H and C57 mice. Strain differences in immune function, however, were observed. CONCLUSIONS: Immune function is unresponsive to photoperiod in these laboratory mice, possibly indicating; a link between immune and reproductive responsiveness to photoperiod, the decreased predictive value of this annual cue to domesticated animals, or both. PMID- 15361816 TI - Bovine pars tuberalis secretions release growth hormone from rat pars distalis of pituitary gland. AB - The pituitary pars tuberalis (PT) is characterized by PT-specific secretory cells which have raised the possibility of an endocrine function for this portion of adenohypophysis. OBJECTIVE: To investigate the effect of the secretion of bovine PT cells into culture medium on growth hormone (GH) response of dispersed pars distalis (PD) cells of rats. METHODS AND RESULTS: 48-hour culture medium of all PT cells at 1 microg of protein concentration induced the greatest GH release from PD cells. After PT cells separation on a discontinuos Percoll gradient and culturing, only the culture medium of cells from 50 and 60% Percoll strength released GH from PD cells. Therefore, cells from 50 and 60% strenght Percoll were cultured together. Only 0.2 mg protein of this culture medium was required to induce the maximal GH release from PD cells, suggesting that these cells could be responsible for producing the factor(s) which affect PD somatotrophe cells. After protein separation by 12% SDS-PAGE of this PT culture medium bands were eluted. The biological activity, measured as ng/ml of GH from PD cells, corresponded to a protein(s) of molecular weight between 45 and 66 kDal. CONCLUSIONS: The results indicate that there is an active proteic factor(s) secreted by the PT that acts upon PD cells to stimulate GH release and that PD could be an effector organ for some secretory product(s) of the PT. PMID- 15361817 TI - Maternal and fetal hypothalamo-pituitary-adrenal axis: different response depends upon the mode of parturition. AB - OBJECTIVES: The changes in corticotropin releasing hormone (CRH), adrenocorticotropin hormone (ACTH), cortisol and dehydroepiandrosterone (DHEA) in maternal and fetal plasma were estimated in two group of women undergoing labour after oxytocin induction, or elective cesarean section to correlate changes in maternal and fetal HPA axis to the different stressor conditions. MATERIALS AND METHODS: Blood was sampled from a maternal peripheral vein two days before labour, during the second stage of labour and on the second postnatal day, and also from umbilical vessels just after delivery. Hormone concentrations were measured by RIA and ELSA methods. The maternal plasma CRH concentration before and after labour was similar in both studied groups. RESULTS: Plasma CRH levels measured before labour in both groups were similar, but during labour after oxytocin infusion declined, and was in group of women with elective cesarean section rose, after the delivery there was no difference between groups. The plasma ACTH prior to the delivery was at the same level in all subjects, during the third stage of labour ACTH rose in the both groups, but was statistically higher in group with oxytocin infusion. The concentrations of cortisol before delivery was similar in both groups, then rose to the same level during the delivery and then declined to the level observed before delivery. The DHEA concentration was similar in both groups, did not change during the labour, there was no difference between its level in the umbilical vein and artery. In the group with oxytocin-induction there was no correlation between maternal CRH and ACTH or cortisol concentration, but such a correlation occurred in the group with elective cesarean section. The concentration of ACTH and DHEA in umbilical artery was similar in both groups but the concentrations of CRH and cortisol in umbilical artery in group of newborns delivered after elective cesarean section were statistically higher. CONCLUSION: The activity of the mother's HPA axis is partially inhibited rather by psychological stressors, than events connected with mode of parturition. The newborn's HPA axis responds in a specific way to mother's HPA-axis. The communication between mother's and newborn's HPA is possibly related to many other factors than placental CRH and ACTH. It is necessary to consider effects of central CRH and ACTH on the fine-tune regulation of HPA axis in the late pregnancy and parturition, not detectable due to the high levels of placental hormones. PMID- 15361818 TI - A new ovarian denervation technique and it's effect on sexual cycle, conception rates and offspring numbers in rats. AB - OBJECTIVES: A novel denervation technique of ovarian was used in rats to investigate its effects on sexual cycle, pregnancy rates and offspring numbers. SETTING AND DESIGN: A total of 34 female albino rats were included. Animals were divided into 3 groups. METHODS: In group 1, ovaries were bilaterally denervated. Animals in group 2 underwent sham operation and group 3 comprised of controls. Sexual cycles in animals were followed by vaginal irrigations. Gestations were determined with parturition of rats. RESULTS: Results indicated that in denervated rats, frequency of estrus decreased and period of estrus increased resulting in a decrease in conception rates and offspring numbers when compared to control rats. MAIN FINDINGS: Although the results were consistent with previous denervation techniques, the operational procedure described herein is simpler and requires no sophisticated equipment, suggesting this to be a method of choice in reproductive physiology studies. CONCLUSION: In this study, we denervated the ovary with a technique other than classical in which the tissues except vascular structures over ovarian suspensory ligament were excised. We found that, estrus count decreased, duration of sexual cycle increased, conception rates and the offspring numbers reduced. This is thought to be a result of blockage of ovarian neural control due to denervation. PMID- 15361819 TI - Treatment of giant paraclinoid aneurysms. Own experience. AB - OBJECTIVES: The incidence of giant aneurysms in the paraclinoid section of the internal carotid artery (ICA) is low. They manifest their occurrence by subarachnoid haemorrhage and neurological and hormonal symptoms, resulting from compression on either the nervous structures or on the pituitary. The treatment of these aneurysms is a challenge for the surgeon, due to a difficult access, high operational risk and when the size of operated malformations is large. MATERIAL AND METHODS: Between the years 1994 and 2001, twenty (20) patients with giant paraclinoid aneurysms of the carotid artery were submitted to: endovascular treatment - 7 patients and to microsurgery - 13 patients. The treatment, following the access to the aneurysm and consisting in short-time (temporary) endovascular occlusion of ICA, was applied in some cases, while trapping was the method of choice in other cases. RESULTS: Good results of the endovascular treatment were obtained in 6 patients and, in one (1) case, satisfactory result was observed. Regarding the whole study group, hormonal symptoms (amenorrhoea) occurred in one case only. Four aneurysms were totally embolized, 1 - almost totally and 2 - partially. Recanalisation was observed in one case after 2 years. In the surgical method, good and very good results were obtained in 10 patients and satisfactory results - in 3 patients. CONCLUSION: Application of either the endovascular method or of the microsurgical method - or of both these methods - used individually in case of a given aneurysm, increases the chances for an effective occlusion with a lower operative risk. PMID- 15361820 TI - The hypothalamic-pituitary-thyroid axis in subjects with subclinical thyroid diseases: the impact of the negative feedback mechanism. AB - OBJECTIVE: To evaluate the hypothalamus-pituitary-thyroid (HPT) axis in patients with subclinical thyroid dysfunction recently reported to have either symptoms or organ involvements with potential morbidity, in order to better differentiate these patients with respect to controls. PATIENTS: Sixtythree patients with subclinical hyperthyroidism (HyperT), 178 normal subjects (EuT) and 106 patients with subclinical hypothyroidism (HypoT) were enrolled; the groups presented normal thyroid hormone (FT4, FT3) levels and, respectively, reduced (HyperT), normal (EuT) and increased (HypoT) TSH levels. The negative feedback was simultaneously evaluated by multiple linear regression. RESULTS: The mean TSH, FT4 and FT3 levels were significantly different in the three groups. The negative correlation between thyroid hormones (FT4 and FT3) and TSH secretion was significant (p<0.001 in HyperT; p<0.01 in EuT; p<0.000001 in HypoT group). FT4 mostly contributed to the negative correlation with TSH. The normal ranges of TSH values was accurately defined on the basis of the regression equation in the EuT group, due to the combining influence of both thyroid hormones (FT3 and FT4). No patient of the HyperT or HypoT group fell inside the range of estimated values of the normal group. CONCLUSIONS: The HPT axis in patients with subclinical hyper- and hypo-thyroidism is significantly modified with respect to normal subjects. The status of the axis, as evaluated by the relationship between the three hormones (FT4, FT3, TSH) together considered, is characteristic of the normal or pathologic condition. A reliable method based on the regression analysis is proposed to correctly evaluate the status of the HPT axis. PMID- 15361821 TI - Brain differentiation and preferred partner characteristics in heterosexual and homosexual men and women. AB - OBJECTIVES: The current study examined and compared the preferred partner characteristics of heterosexual and homosexual men and women in relation to speculated patterns of brain differentiation underlying the preferences. Further, the study compared the preferences of butch versus femme homosexual women. METHODS: Two hundred twelve heterosexual and homosexual men and women completed questionnaires on which they rated themselves and their idealized sexual partners on various morphological and behavioral characteristics. RESULTS: Results of within-subjects multivariate analyses of variance (MANOVAs) showed that the pattern of preferred partner characteristics of heterosexual women is most consistent with the theorized brain differentiation underlying those preferences. There was varying support for the theory as it applies to the other three groups. Between-subjects MANOVAs provided evidence to support some of the predictions generated by theory on the differences in brain differentiation between heterosexual and homosexual men and women and between homosexual women categorized as butch and femme. CONCLUSION: The overall pattern of preferred partner characteristics among and between heterosexual and homosexual men and women does not support theory that holds that underlying brain differentiation between the groups is discrete and categorical. Rather, it supports theory that holds that differentiation manifests itself on a continuum leading to a variety of patterns of sexual orientation, and by extension, preferred partner characteristics. PMID- 15361822 TI - C-reactive protein as a predictor of threatening preterm delivery. AB - OBJECTIVES: Preterm deliveries represent still one of the most important problems in contemporary obstetrics. They are associated with prematurity and higher rate of perinatal mortality and morbidity. During the last few years, the role of C reactive protein (CRP) in prediction of threatening preterm delivery was emphasized. CRP is produced mainly inside the liver as a response to acute and chronic inflammatory processes. AIM: The aim of this study was to assess relations between C-reactive protein, oxytocinase, izooxytocinase and vaginal culture in prediction of preterm delivery. METHODS: This study was performed in the years 2000-2004 in the Department of Septic Obstetrics and Gynecology of Collegium Medicum of the Jagiellonian University. Some 389 patients hospitalized because of threatening preterm delivery or preterm delivery in tractu were enrolled into the trial. CONCLUSION: C-reactive protein is a useful marker of threatening preterm delivery, overtaking the results of vaginal culture. CAP1 and CAP2 are effective biochemical markers in pregnancy monitoring. PMID- 15361823 TI - Coexisting acromegaly and primary empty sella syndrome. AB - The normal functions of the pituitary gland may be suppressed when the gland is compressed onto the sella floor by arachnoid tissue extending through an impaired sella diaphragm. Interestingly, pituitary hormone hypo- and hypersecretion, including acromegaly, have been observed in patients with an 'empty sella'(1-4). This 'empty sella syndrome' has been classified into a primary form, in which no inciting factor (pituitary irradiation or surgery for a pituitary tumor) is present, and a secondary form, in which the empty sella occurs after pituitary procedures. In this report we describe a patient who presented with clinical and biochemical features of acromegaly and who had an empty sella on pituitary magnetic resonance imaging (MRI). PMID- 15361824 TI - Zhennan Xue - ad multos transannos. PMID- 15361825 TI - Human Arf tumor suppressor specifically interacts with chromatin containing the promoter of rRNA genes. AB - The tumor suppressor Arf (Alternative Reading Frame) protein (p14ARF in human and p19ARF in mouse) is mainly located in the nucleolus consistent with its subcellular localization, the protein has been shown to specifically interact with 5.8S rRNA and with B23/Nucleophosmin and to regulate ribosome biogenesis. Here, we show that the p14ARF protein interacts with chromatin and is recovered by chromatin immunoprecipitation (ChIP) in a fraction that contains a DNA sequence of the rRNA gene promoter. In addition, topoisomerase I (Topo I) that has been shown to interact with p14ARF coprecipitates with p14ARF containing chromatin. These data, in view of the function for Topo I in rRNA transcription, are consistent with a role for the p14ARF-Topo I complex in rRNA transcription and/or maturation. PMID- 15361826 TI - Cooperation of betulinic acid and TRAIL to induce apoptosis in tumor cells. AB - We previously reported that the TRAIL (tumor necrosis factor (TNF)-related apoptosis-inducing ligand)-induced death signal requires amplification by mitochondria in certain cell types, for example, in type II cells. Here, we provide for the first time evidence that the natural compound betulinic acid (BetA) cooperated with TRAIL to induce apoptosis in tumor cells. Through functional complementation, simultaneous stimulation of the death receptor pathway by TRAIL and the mitochondrial pathway by BetA resulted in complete activation of effector caspases, apoptosis and inhibition of clonogenic survival. BetA and TRAIL cooperated to trigger loss of mitochondrial membrane potential and release of cytochrome c and Smac from mitochondria. Also, combination treatment with BetA and TRAIL resulted in increased cleavage of caspase-8 and Bid indicating that activation of effector caspases may feed back in a positive amplification loop. Importantly, the combination treatment with BetA and TRAIL cooperated to induce apoptosis in different tumor cell lines and also in primary tumor cells, but not in normal human fibroblasts indicating some tumor specificity. Since most human cancers represent type II cells, triggering the mitochondrial pathway by BetA may be a novel approach to enhance the efficacy of TRAIL-based therapies, which warrants further investigation. PMID- 15361827 TI - Differentially expressed genes execute zinc-induced apoptosis in precancerous esophageal epithelium of zinc-deficient rats. AB - Zinc deficiency (ZD) in rats increases esophageal cell proliferation and the incidence of N-nitrosomethylbenzylamine-induced esophageal tumors. Conversely, zinc replenishment (ZR) rapidly induces apoptosis in esophageal epithelia and reverses cancer development. We investigated gene expression changes in ZR versus ZD esophageal epithelia to identify differentially expressed genes associated with the antitumor effect of ZR. Weanling rats were fed a ZD diet for 6 weeks to establish esophageal cell proliferation or a zinc-sufficient (ZS) diet. Then, 10 ZD rats were treated with zinc gluconate intragastrically and switched to ZS diet; the remaining 10 ZD and ZS animals were treated with saline. All animals were killed 26-28 h later. Using cDNA microarrays, real-time polymerase chain reaction amplification and RNA hybridization techniques, we identified novel differentially expressed genes, including a RNA-binding protein with two RNA recognition motifs and a zinc knuckle (ZD7), and a DNA/RNA helicase with a DEAD box (ZD10) with two splice variants, ZD10a and ZD10b. In situ hybridization detected increased mRNA expression of ZD7, ZD10a and ZD10b in ZR esophageal epithelia, which displayed markedly increased occurrence of apoptotic cells, relative to ZD epithelia. Overexpression of ZD7 in human esophageal cancer cells resulted in induction of apoptosis and activation of caspase-3 and -7, activities that were inhibited by caspase-specific inhibitors. In addition, ZD7 mRNA levels and zinc-induced apoptosis in rat squamous carcinoma cells were reduced by specific small interfering ribonucleic acids. Thus, ZR rapidly induces ZD7 and ZD10 expression, which in turn stimulates apoptosis. These results provide the beginnings of a molecular pathway for zinc-induced apoptosis under conditions that reverse esophageal tumor initiation. PMID- 15361828 TI - Induction of PDCD4 tumor suppressor gene expression by RAR agonists, antiestrogen and HER-2/neu antagonist in breast cancer cells. Evidence for a role in apoptosis. AB - The growth of human breast tumor cells is regulated through signaling involving cell surface growth factor receptors and nuclear receptors of the steroid/thyroid/retinoid receptor gene family. Retinoic acid receptors (RARs), members of the steroid/thyroid hormone receptor gene family, are ligand-dependent transcription factors, which have in vitro and in vivo growth inhibitory activity against breast cancer cells. RAR-agonists inhibit the proliferation of many human breast cancer cell lines, particularly those whose growth is stimulated by estradiol (E2) or growth factors. Additionally, RAR-agonists and synthetic retinoids such as Ferentinide have been shown to induce apoptosis in malignant breast cells but not normal breast cells. To better define the genes involved in RAR-mediated growth inhibition of breast cancer cells, we used oligonucleotide microarray analysis to create a database of genes that are potentially regulated by RAR-agonists in breast cancer cells. We found that PDCD4 (programmed cell death 4), a tumor suppressor gene presently being evaluated as a target for chemoprevention, was induced about three-fold by the RARalpha-selective agonist Am580, in T-47D breast cancer cells. RAR pan-agonists and Am580, but not retinoid X receptors (RXR)-agonists, stimulate the expression of PDCD4 in a wide variety of retinoid-inhibited breast cancer cell lines. RAR-agonists did not induce PDCD4 expression in breast cancer cell lines, which were not growth inhibited by retinoids. We also observed that antiestrogen and the HER-2/neu antagonist, Herceptin (Trastuzumab), also induced PDCD4 expression in T-47D cells, suggesting that PDCD4 may play a central role in growth inhibition in breast cancer cells. Transient overexpression of PDCD4 in T-47D (ER+, RAR+) and MDA-MB-231 (ER-, RAR-) cells resulted in apoptotic death, suggesting a role for PDCD4 in mediating apoptosis in breast cancer cells. PDCD4 protein expression has previously been reported in small ductal epithelium of normal breast. To date, there has been no report of induction of PDCD4 expression by RAR-agonists, antiestrogen or HER2/neu antagonist in breast cancer cells and its potential role in apoptosis in these cells. PMID- 15361829 TI - Budesonide exerts its chemopreventive efficacy during mouse lung tumorigenesis by modulating gene expressions. AB - Budesonide, a glucocorticoid, was proven to be a highly effective agent in preventing the development of lung tumors in A/J mice. In a lung tumor bioassay, budesonide produced 70% inhibition of tumor multiplicity and 94% reduction of total tumor load compared to benzopyrene (B[a]P) treated mice. Gene expression array analysis was performed on mouse lung tumors from this bioassay using Affymetrix U74Av2 GeneChips to determine gene expression changes associated with budesonide treatment. We found 363 genes that were changed between lung tumors induced by treatment with B[a]P and similar tumors treated with budesonide. Among them, 243 genes were overexpressed and 120 genes were underexpressed after budesonide treatment. In addition, 108 genes differentially expressed during mouse lung tumorigenesis (50 genes overexpressed and 58 genes underexpressed) were modulated back to normal levels after budesonide treatment when compared with the controls group. These genes are involved in a broad range of different pathways including control of cell cycle, signal transduction, and apoptosis and may play a role in the observed preventive effect. Our results suggest that budesonide exerts its effects of chemoprevention through growth arrest via Mad2/3 and through apoptosis via Bim/Blk and, by inference, caspase-8/9. Using the pathway visualization tool GenMapp, G protein pathway and MAPK cascade were also regulated by budesonide. Thus, we have determined, for the first time, the expression profiles of genes modulated by budesonide during murine lung tumorigenesis. Our results indicate that the chemopreventive effects of budesonide in the mouse lung tumorigenesis assay involved increase and decrease expression of a wide variety of genes in multiple signaling pathways. PMID- 15361830 TI - Activation of ATM and Chk2 kinases in relation to the amount of DNA strand breaks. AB - The diverse checkpoint responses to DNA damage may reflect differential sensitivities by molecular components of the damage-signalling network to the type and amount of lesions. Here, we determined the kinetics of activation of the checkpoint kinases ATM and Chk2 (the latter substrate of ATM) in relation to the initial yield of genomic DNA single-strand (SSBs) and double-strand breaks (DSBs). We show that doses of gamma-radiation (IR) as low as 0.25 Gy, which generate vast numbers of SSBs but only a few DSBs per cell (<8), promptly activate ATM kinase and induce the phosphorylation of the ATM substrates p53 Ser15, Nbs1-Ser343 and Chk2-Thr68. The full activation of Chk2 kinase, however, is triggered by treatments inflicting >19 DSBs per cell (e.g. 1 Gy), which cause Chk2 autophosphorylation on Thr387, Chk2-dependent accumulation of p21waf1 and checkpoint arrest in the S phase. Our results indicate that, in contrast to ATM, Chk2 activity is triggered by a greater number of DSBs, implying that, below a certain threshold level of lesions (<19 DSBs), DNA repair can occur through ATM, without enforcing Chk2-dependent checkpoints. PMID- 15361831 TI - Survivin regulates the p53 tumor suppressor gene family. AB - Survivin regulates cell division and inhibits apoptosis by blocking caspase activation. The tumor suppressor p53 inhibits cell cycle progression and induces apoptosis. Since Survivin overexpression and loss of wild-type p53 expression/function occur in most cancers, we investigated whether Survivin regulates p53. Stable overexpression of Survivin protects BaF3 cells from Adriamycin-induced apoptosis, while dominant-negative (T34A) and antisense (AS) Survivin accelerate apoptosis. In BaF3 cells and transiently transfected MCF7 breast cancer cells, elevation of total and phospho-Ser15-p53 in response to Adriamycin is blocked by Survivin and enhanced by Survivin disruption. Furthermore, in Adriamycin-treated MCF7 cells, ectopic Survivin decreased p53 mRNA and increased mRNA and protein of the p53 homologues DeltaNp63 and TAp73 and mRNA for DeltaNp73, suggesting that Survivin may differentially regulate p53 family transcription. Concomitant with decreasing p53 mRNA, Survivin decreased Mdm2 mRNA. Survivin disruption by T34A or AS Survivin resulted in reduced Mdm2 protein. The caspase inhibitor, Z-VAD-FMK, blocked the decrease in Mdm2 as well as the increase in p53 resulting from Survivin disruption, indicating that Survivin regulates Mdm2 at the post-translational level. Proteosome inhibition confirmed that reduced p53 protein observed in cells overexpressing Survivin is due to enhanced p53 degradation resulting from Survivin-mediated inhibition of Mdm2 cleavage by caspases. In summary, our results identify regulatory interactions between Survivin and p53 at the mRNA and protein levels, and suggest that the p53 homologues DeltaNp63, TAp73 and DeltaNp73 may also be regulated by Survivin. PMID- 15361832 TI - KLF2 inhibits Jurkat T leukemia cell growth via upregulation of cyclin-dependent kinase inhibitor p21WAF1/CIP1. AB - Kruppel-like factor 2 (KLF2) is a member of the KLF family of zinc-finger transcription factors and is involved in maintaining T-cell quiescence, regulating preadipocyte differentiation, endothelial cell function and lung development. We used a tetracycline-inducible system in Jurkat T leukemia cells to study the biological role of KLF2 in cellular growth and differentiation. Our results show that expression of KLF2 inhibits cell growth in autonomously proliferating Jurkat cells. Further, 3H-thymidine uptake assays indicate that KLF2 inhibits DNA synthesis in these cells. Moreover, both activation and inhibitory domains are required for KLF2 to suppress Jurkat cell proliferation. In addition, KLF2 upregulates p21WAF1/CIP1 expression. Additionally, we found that KLF2 upregulates p21WAF1/CIP1 promoter activity in Jurkat, HepG2 and SW480 cells. Our analysis shows that the potential KLF2 responsive elements are located between -124 and -60 of the p21WAF1/CIP1 promoter. The sole CACCC site, a sequence recognized by KLF2, in this region is not the element responsive to KLF2. Finally, we determined that the Sp1-3-binding site is the functional responsive element of KLF2 in the p21WAF1/CIP1 promoter, and we conclude that KLF2 directly regulates p21WAF1/CIP1 expression. PMID- 15361833 TI - The regulation of CHK2 in human cancer. AB - Exceptional progress has been made in the past two decades in mapping oncogenes and tumour suppressors, defining a function for these master switches, and identifying novel anti-cancer drug targets. The p53 tumour suppressor is a central component of a DNA-damage-inducible pathway controlled by the ataxia telangiectasia mutated (ATM) and CHK2 protein kinases that have a central role in cancer suppression. One limitation of current human cancer research is the difficulty in developing genetic models that reveal the post-translational regulation of a growth suppressor like CHK2 within the microenvironment of a human tumour. Gaining such insights is important since yeast models and human tissue culture cell lines do not necessarily predict how enzymes like CHK2 are regulated in vivo, and therefore what factors can affect CHK2 tumour suppressor function. Translational cancer research aims to link basic research methodologies and clinical biology by uncovering cancer-specific pathways not revealed by other approaches. This approach is exemplified by two studies in this edition of Oncogene: both use a set of well-characterized human cancers with the objective of identifying novel post-translational control of the tumour suppressor CHK2. The authors have revealed two unexpected epigenetic modifications of the CHK2 pathway in vivo: (1) constitutive phosphorylation of CHK2 at its ATM-activated site in the absence of exogenous DNA damage; and (2) the production of hyper spliced and inactive isoforms of CHK2. These studies highlight the need to develop model systems to understand why CHK2-activating pathways are being triggered or suppressed in different human cancers and whether the splicing machinery can be manipulated to control the activity of CHK2 for therapeutic benefit. PMID- 15361834 TI - ICBP90, an E2F-1 target, recruits HDAC1 and binds to methyl-CpG through its SRA domain. AB - ICBP90, inverted CCAAT box-binding protein of 90 kDa, has been reported as a regulator of topoisomerase IIalpha expression. We present evidence here that ICBP90 binds to methyl-CpG when at least one symmetrically methylated-CpG dinucleotides is presented as its recognition sequence. A SET and RING finger associated (SRA) domain accounts for the high binding affinity of ICBP90 for methyl-CpG dinucleotides. This protein constitutes a complex with HDAC1 also via its SRA domain, and bound to methylated promoter regions of various tumor suppressor genes, including p16INK4Aand p14ARF, in cancer cells. It has been reported that expression of ICBP90 was upregulated by E2F-1, and we confirmed that the upregulation was caused by binding of E2F-1 to the intron1 of ICBP90, which contains two E2F-1-binding motifs. Our data also revealed accumulation of ICBP90 in breast-cancer cells, where it might suppress expression of tumor suppressor genes through deacetylation of histones after recruitment of HDAC1. The data reported here suggest that ICBP90 is involved in cell proliferation by way of methylation-mediated regulation of certain genes. PMID- 15361835 TI - Identification of cell surface and secreted proteins essential for tumor cell survival using a genetic suppressor element screen. AB - Survival factors play critical roles in regulating cell growth in normal and cancer cells. We designed a genetic screen to identify survival factors which protect tumor cells from apoptosis. A retroviral expression library of random cDNA fragments was constructed from cancer cells and used to transduce the colon carcinoma cell line HCT116. Recipient cells were functionally selected for induction of caspase 3-mediated apoptosis. Analyses of over 10,000 putative genetic suppression elements (GSEs) sequences revealed cognate gene candidates that are implicated in apoptosis. We further analysed 26 genes encoding cell surface and secreted proteins that can potentially serve as targets for therapeutic antibodies. Tetracycline-inducible GSEs from several gene candidates induced apoptosis in stable HCT 116 cell lines. Similar phenotypes were caused by RNAi derived from the same genes. Our data suggest requirement for the cell surface targets IGF2R, L1CAM and SLC31A1 in tumor cell growth in vitro, and suggests that IGF2R is required for xenograft tumor growth in a mouse model. PMID- 15361836 TI - Effects of a conditionally active v-ErbB and an EGF-R inhibitor on transformation of NIH-3T3 cells and abrogation of cytokine dependency of hematopoietic cells. AB - Epidermal growth factor (EGF) and its cognate receptor (EGF-R) are often dysregulated in human neoplasia. Moreover, EGF-R-transformed cell lines have constitutive EGF-R activity, which makes elucidation of its effects difficult to determine. In the following studies, the effects of a novel conditionally activated form of EGF-R, v-ErbB:ER, on the morphological transformation of NIH 3T3 cells and the abrogation of hematopoietic cell cytokine dependence were investigated. The v-ErbB ES-4 oncogene was fused to the hormone binding domain of the estrogen receptor (ER). This construct, v-ErbB:ER, requires beta-estradiol or 4-OH tamoxifen for activation. v-ErbB:ER conditionally transformed NIH-3T3 cells and abrogated cytokine dependence of hematopoietic cells. Stimulation of v ErbB:ER activity resulted in the activation of the phosphatidylinositol 3-kinase (PI3K)/Akt and Raf/MEK/ERK kinase cascades. To determine the importance of these signal transduction pathways, the conditionally transformed hematopoietic cells were treated with EGF-R, PI3K and MEK inhibitors. The EGF-R inhibitor AG1478 effectively inhibited MEK, ERK and Akt activation, and induced apoptosis when the cells were grown in response to v-ErbB:ER. Apoptosis was observed at 100- to 1000 fold lower concentrations of AG1478 when the cells were grown in response to v ErbB:ER as opposed to IL-3. Furthermore, the parental, BCR-ABL- and Raf transformed cells were only susceptible to the apoptosis-inducing effects of AG1478 at the highest concentrations demonstrating the specificity of these inhibitors. MEK or PI3K inhibitors suppressed ERK or Akt activation, respectively, and induced apoptosis in the v-ErbB:ER-responsive cells. However, MEK and PI3K inhibitors only induced apoptosis at 1000-fold higher concentrations than the EGFR inhibitor. This novel v-ErbB:ER construct and these conditionally transformed cell lines will be useful to further elucidate ErbB-mediated signal transduction and to determine the effectiveness of various inhibitors in targeting different aspects of EGF-R-mediated signal transduction and malignant transformation. PMID- 15361837 TI - Inhibition of glycogen synthase kinase-3 represses androgen receptor activity and prostate cancer cell growth. AB - The transcriptional activity of the androgen receptor (AR) is regulated by interaction with various coregulators, one of which is beta-catenin. Interest in the role of beta-catenin in prostate cancer has been stimulated by reports showing that it is aberrantly expressed in the cytoplasm and/or nucleus in up to 38% of hormone-refractory tumours and that overexpression of beta-catenin results in activation of AR transcriptional activity. We have examined the effect of depleting endogenous beta-catenin on AR activity using Axin and RNA interference. Axin, which promotes beta-catenin degradation, inhibited AR transcriptional activity. However, this did not require the beta-catenin-binding domain of Axin. Depletion of beta-catenin using RNA interference increased, rather than decreased, AR activity, suggesting that endogenous beta-catenin is not a transcriptional coactivator for the AR. The glycogen synthase kinase-3 (GSK-3) binding domain of Axin prevented formation of a GSK-3-AR complex and was both necessary and sufficient for inhibition of AR-dependent transcription. A second GSK-3-binding protein, FRAT, also inhibited AR transcriptional activity, as did the GSK-3 inhibitors SB216763 and SB415286. Finally, inhibition of GSK-3 reduced the growth of AR-expressing prostate cancer cell lines. Our observations suggest a potential new therapeutic application for GSK-3 inhibitors in prostate cancer. PMID- 15361838 TI - Recombinant CD44-HABD is a novel and potent direct angiogenesis inhibitor enforcing endothelial cell-specific growth inhibition independently of hyaluronic acid binding. AB - CD44 is the main cellular receptor for hyaluronic acid (HA). We previously found that overexpression of CD44 inhibited tumor growth of mouse fibrosarcoma cells in mice. Here, we show that soluble recombinant CD44 HA-binding domain (CD44-HABD) acts directly onto endothelial cells by inhibiting endothelial cell proliferation in a cell-specific manner. Consequently, soluble recombinant CD44-HABD also blocked angiogenesis in vivo in chick and mouse, and thereby inhibited tumor growth of various origins at very low doses (0.25 mg/kg x day). The antiangiogenic effect of CD44 is independent of its HA-binding capacity, since mutants deficient in HA binding still maintain their antiangiogenic and antiproliferative properties. Recombinant CD44-HABD represents a novel class of angiogenesis inhibitors based on a cell-surface receptor. PMID- 15361839 TI - Mutant ras-induced proliferation of human thyroid epithelial cells requires three effector pathways. AB - Ras mutations occur as an early event in many human tumours of epithelial origin, including thyroid. Using primary human thyroid epithelial cells to model tumour initiation by Ras, we have shown previously that activation of both the MAP kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) effector pathways are necessary, but even when activated together are not sufficient, for Ras-induced proliferation. Here, we show that a third effector, RalGEF, is also activated by Ras in these cells, that this activation is necessary for Ras-induced proliferation, and furthermore that in combination with the MAPK and PI3K effectors, it is able to reproduce the proliferative effect of activated Ras. The requirement for three effector pathways indicates a more robust control of cell proliferation in this normal human epithelial cell type than has been displayed in previous similar studies using rodent and human cell lines. Our findings highlight the importance of the appropriate cellular context in models of Ras induced tumour development. PMID- 15361840 TI - Mutation of GATA3 in human breast tumors. AB - GATA3 is an essential transcription factor that was first identified as a regulator of immune cell function. In recent microarray analyses of human breast tumors, both normal breast luminal epithelium and estrogen receptor (ESR1) positive tumors showed high expression of GATA3. We sequenced genomic DNA from 111 breast tumors and three breast-tumor-derived cell lines and identified somatic mutations of GATA3 in five tumors and the MCF-7 cell line. These mutations cluster in the vicinity of the highly conserved second zinc-finger that is required for DNA binding. In addition to these five, we identified using cDNA sequencing a unique mis-splicing variant that caused a frameshift mutation. One of the somatic mutations we identified was identical to a germline GATA3 mutation reported in two kindreds with HDR syndrome/OMIM #146255, which is an autosomal dominant syndrome caused by the haplo-insufficiency of GATA3. The ectopic expression of GATA3 in human 293T cells caused the induction of 73 genes including six cytokeratins, and inhibited cell line doubling times. These data suggest that GATA3 is involved in growth control and the maintenance of the differentiated state in epithelial cells, and that GATA3 variants may contribute to tumorigenesis in ESR1-positive breast tumors. PMID- 15361841 TI - Identification of STAG1 as a key mediator of a p53-dependent apoptotic pathway. AB - A mutant version of p53 (p53-121F), in which phenylalanine replaces the 121st serine residue, can induce apoptosis more effectively than wild-type p53 (wt p53). In view of this observation, we considered that one or more apoptosis related p53-target genes might be preferentially induced by p53-121F. We carried out cDNA microarray analysis to identify such genes, using mRNAs isolated from LS174T colon-cancer cells infected by adenovirus vectors containing either p53 121F (Ad-p53-121F) or wt-p53 (Ad-p53). The STAG1 gene was one of the transcripts showing higher expression levels in cells infected with Ad-p53-121F as opposed to Ad-wtp53. The encoded product appears to contain a transmembrane domain, and binding motifs for SH3 and WW. In two other cancer cell lines, the expression of STAG1 mRNA was induced in response to various genotoxic stresses in a p53 dependent manner; moreover, enforced expression of STAG1 led to apoptosis in several additional cancer cell lines. Suppression of endogenous STAG1 using the RNA-interference method reduced the apoptotic response, whether induced by Ad-p53 121F or Ad-p53. These results suggest that STAG1, a novel transcriptional target for p53, mediates p53-dependent apoptosis, and might be a good candidate for next generation gene therapy. PMID- 15361842 TI - RARbeta2 is a candidate tumor suppressor gene in myelofibrosis with myeloid metaplasia. AB - Myelofibrosis with myeloid metaplasia (MMM) is a clonal stem-cell disorder that leads to ineffective hematopoiesis, bone marrow fibrosis, and extramedullary hematopoiesis. The molecular mechanisms underlying the development of this myeloproliferative syndrome are currently unknown. In order to identify tumor suppressor genes that may be involved in the disease process, we performed an analysis for loss of heterozygosity (LOH) in CD34+ cells from 29 patients with MMM. We observed a frequency of allelic loss on chromosomal arm 3p in 24% of cases. Detailed mapping of 3p revealed a distinct region of deletion at 3p24. Among the genes known to map within this region is the retinoic acid receptor beta (RARbeta2) gene. To determine whether RARbeta2 gene activity is diminished in this disease, we analysed its expression in CD34+ cells from 17 patients with MMM using quantitative PCR. Our results indicate that expression of RARbeta2 is significantly decreased in 100% of patient samples compared to that in CD34+ cells from 10 normal individuals. Since allelic loss at 3p24 occurs in <25% of patients, we investigated the contribution of epigenetic modifications to RARbeta2 inactivity. Using methylation-specific PCR, we found hypermethylation of RARbeta2 in 16 of 18 patients (89%), while the methylated form of the gene was absent in CD34+ cells from nine normal individuals. Our results suggest that RARbeta2 acts as a tumor suppressor gene in MMM and that epigenetic changes are the most significant determinants of RARbeta2 gene activity in these patients. PMID- 15361843 TI - Differential hypermethylation of SOCS genes in ovarian and breast carcinomas. AB - Suppressor of cytokine signaling (SOCS) proteins have emerged as critical attenuators of cytokine-mediated processes, suggesting a role in the suppression of tumorigenesis. In the ovary and mammary gland, cytokines such as prolactin and IL-6 are important regulators of growth and differentiation. We have investigated whether silencing or inactivation of SOCS genes occurs in ovarian and breast carcinomas. The SOCS1 and SOCS2 CpG islands were found to be hypermethylated in 23 and 14% of primary ovarian cancers, respectively, whereas only SOCS1 was methylated in breast cancers (9%). Methylation of these genes did not occur in normal tissues. No correlation was apparent between methylation and loss of heterozygosity, and no somatic mutations were found in a large panel of carcinomas. Aberrant methylation of these SOCS genes correlated with transcriptional silencing in ovarian and breast cancer cell lines, since expression was induced by the demethylating agent 5-azadeoxycytidine. SOCS3 was not hypermethylated in either cancer type. Consistent with this data, SOCS1 and SOCS2 but not SOCS3 suppressed the growth of ovarian and breast cancer cells. Hypermethylation and silencing of specific SOCS genes in the ovary, and to a lesser extent in breast, may augment cytokine responsiveness in these tissues, thereby contributing to oncogenesis. PMID- 15361844 TI - Mutation at p53 serine 389 does not rescue the embryonic lethality in mdm2 or mdm4 null mice. AB - Mdm2 and its homolog Mdm4 inhibit the function of the tumor suppressor p53. Targeted disruption of either mdm2 or mdm4 genes in mice results in embryonic lethality that is completely rescued by concomitant deletion of p53, suggesting that deletion of negative regulators of p53 results in a constitutively active p53. Thus, these mouse models offer a unique in vivo system to assay the functional significance of different p53 modifications. Phosphorylation of serine 389 in murine p53 occurs specifically after ultraviolet-light-induced DNA damage, and phosphorylation of this site enhances p53 activity both in vitro and in vivo. Recently, mice with a serine to alanine substitution at serine 389 (p53S389A) in the endogenous p53 locus were generated. To examine the in vivo significance of serine 389 phosphorylation during embryogenesis, we crossed these mutant mice to mice lacking mdm2 or mdm4. The p53S389A allele did not alter the embryonic lethality of mdm2 or mdm4. Additional crosses to assay the effect of one p53S389A allele with a p53 null allele also did not rescue the lethal phenotypes. In conclusion, the phenotypes due to loss of mdm2 or mdm4 were not even partially rescued by p53S389A, suggesting that p53S389A is functionally wild type during embryogenesis. PMID- 15361845 TI - A novel p21WAF1/CIP1 transcript is highly dependent on p53 for its basal expression in mouse tissues. AB - p21WAF1/CIP1 is an important transcriptional target of p53 and it plays a critical role in growth arrest after DNA damage. Here, we report the identification of a novel alternate mouse p21 transcript that is conserved in evolution. It differs from the classical p21WAF1/CIP1 transcript in the first exon, which is located at approximately 2.8 kb upstream of transcriptional start site of p21WAF1/CIP1 and is sandwiched between two p53 binding sites. This novel p21 transcript is present in most mouse tissues with highest levels of expression in the spleen. In contrast to the classical p21WAF1/CIP1 transcript, this new transcript is highly dependent on p53 for its basal expression, as evidenced by its absence in nearly all of p53-/- mouse tissues. This transcript is also absent at nonpermissive temperature in a 10-1 mouse cell line lacking endogenous p53 and harboring temperature-sensitive p53 mutant. However, this novel transcript is induced to appreciable levels in the presence of high p53 activity at the permissive temperature. Our data suggest that p53-dependent induction of p21 may be an additive effect conferred by individual increases in the alternate and classical p21 transcripts. PMID- 15361846 TI - Involvement of p53 and Fas/CD95 in murine neural progenitor cell response to ionizing irradiation. AB - We investigated the role of tumor suppressor p53 and Fas (CD95/APO-1), a member of the tumor necrosis factor receptor family, in neural progenitors response to gamma-irradiation exposure. Telencephalic cells were obtained from wild-type C57Bl/6, or p53-/- or fas-/-, 15-day-old mouse embryos. They were cultured in conditions allowing neural progenitors to form proliferating clusters (neurospheres). A 2 Gy gamma-irradiation induced a G1 cell cycle arrest and triggered apoptosis in wild-type neural progenitor cultures in correlation with an enhanced expression of p53 and of its downstream target p21(WAF1), both of them acquiring a nuclear localization. These effects did not occur in p53-/- neural progenitors demonstrating the central role played by p53 in their response to ionizing radiation. Furthermore, the monoclonal antibody Jo2 directed against Fas induced apoptosis of wild type but not of fas-/- neural progenitors, indicating the existence of a functional Fas signaling pathway in neural progenitors. Ionizing radiation induced an increase of Fas membrane expression related to a p53-dependent increase of fas mRNA expression in wild-type neural progenitors. Moreover, fas-/- neural progenitors exhibited delayed radiation induced apoptosis compared to wild-type cells. Therefore, these findings establish a role for Fas/CD95 related to p53 in the response of neural progenitors to gamma-radiation exposure. Similar mechanisms could be triggered in neural progenitors in case of different stresses during brain development or in the course of various diseases affecting the adult brain. PMID- 15361847 TI - HIV-Tat promotes cellular proliferation and inhibits NGF-induced differentiation through mechanisms involving Id1 regulation. AB - Id1 is a helix-loop-helix transcriptional factor that controls growth and survival of neuronal cells. Downregulation of Id1 expression is required to initiate differentiation and cell-cycle withdrawal in primary neuronal culture as well as in PC12 cells. The HIV-1 transactivating factor, Tat, has been suspected of causing neuronal dysfunction that often leads to the development of HIV associated dementia in AIDS patients. We found that the expression of Tat in PC12 cells promotes serum-independent growth, formation of large colonies in soft agar, and the acceleration of tumor growth in nude mice. In addition, Tat showed the ability to inhibit the nerve growth factor (NGF)-induced neuronal differentiation of PC12 cells. Our results show that the Tat-mediated signaling events, which lead to serum-independent growth and the inhibition of NGF-induced differentiation, have a common cellular target: the upregulation of Id1 expression. In the absence of NGF, expression of Id1 is required to promote serum independent proliferation of PC12/Tat cells, as the inhibition of Id1 by antisense DNA restored the serum-dependent growth of PC12/Tat cells. In the presence of NGF, Tat utilizes an additional pathway that involves phosphorylation of Stat5a, to upregulate Id1 expression and block neuronal cell differentiation. Suppression of Stat5a by use of its dominant-negative mutant reversed the transient expression of Id1 and the blockage of NGF-mediated differentiation in PC12/Tat cells. Finally, the treatment of PC12 cells with recombinant Tat also enhanced the NGF-induced Id1 expression, further pointing to Id1 as a target for Tat. Taken together, these studies suggest additional targets for Tat action in neuronal cells and provide new insights into the mechanisms involved in the dysregulation of neuronal functions. PMID- 15361848 TI - Checkpoint genes, ageing, and the development of cancer. AB - Mathematical models are used to examine the relationship between checkpoint competence, ageing, and the development of cancer. The models take into account the dynamics of healthy tissue, the dynamics of initial tumor growth, and the interactions between healthy tissue and tumor cells. Two types of behavior are found. (i) A reduction of checkpoint competence results in reduced ageing of tissue, but in faster development and progression of tumors. (ii) Reduced checkpoint competence results both in reduced ageing of tissue, and in a reduced incidence of tumors. The tumors which do become established, however, are predicted to progress at an accelerated rate. The models define the conditions under which this counter-intuitive finding is observed. One reason could be the relationship between checkpoint activity and the ability of the tissue environment to exert inhibitory effects on tumor cells. Checkpoints induce senescence in tissue cells, and this compromises their ability to suppress tumor growth. Reduced checkpoint competence leads to reduced tissue senescence, and this results in higher amounts of tumor inhibition. The theoretical work is discussed with respect to data from p53 mutant mice, which show both types of relationships suggested by the models. The models help to identify differences in the experimental protocols which could explain the seemingly contradictory observations. PMID- 15361849 TI - Dose-dependent effect of FHIT-inducible expression in Calu-1 lung cancer cell line. AB - Abnormalities in the expression of the tumour suppressor fragile histidine triad (FHIT) gene have been reported in a variety of human tumours, including lung cancer and restoration of its expression in cancer cell lines resulted in the inhibition of proliferation and apoptosis induction. Most of the studies that have assigned a proapoptotic role to the FHIT gene were performed in adenoviral FHIT-transduced cancer cells expressing high levels of the Fhit protein. The present work was the first study designed to investigate the effects of FHIT gene replacement in a human FHIT-negative non-small-cell lung cancer (NSCLC) cell line (Calu-1) by using a hormone-inducible expression system that allows tight modulation of the transgene expression. Through this approach, we demonstrated that a prolonged induction was required to accumulate the Fhit protein at levels adequate to promote a significant decrease of cell proliferation. Analysis of cell-cycle phase distribution showed an accumulation of cells in the G0/G1 phase and a concomitant decrease in the S phase. Moreover, an upregulation of p21waf1 transcript was found, which could account for the alteration of the cycling properties of the cells. The growth-inhibitory effects observed were not associated with apoptosis appearance, and although in these conditions the Fhit protein content was higher than in normal bronchial human epithelial cells (NHBE), it was still significantly lower than the level capable of inducing apoptosis in Calu-1 cells after adenoviral-mediated FHIT gene transfer. These results indicate that the tumour suppressor properties of Fhit are strictly related to its expression level and show that the Fhit protein has a dose dependent antiproliferative effect on the Fhit-negative Calu-1 lung cancer cell line. PMID- 15361850 TI - Aggrus: a diagnostic marker that distinguishes seminoma from embryonal carcinoma in testicular germ cell tumors. AB - Aggrus (also known as T1alpha/podoplanin) is a membrane sialoglycoprotein whose function in tumors is unknown. We recently determined that Aggrus possessed the ability of inducing platelet aggregation and that its expression was frequently upregulated in colorectal tumors. Thus, Aggrus expression might be associated with tumor-induced platelet aggregation and tumor metastasis. Here we show, by means of cancer profiling array and real-time PCR, that aggrus mRNA expression is frequently upregulated in testicular germ cell tumors when compared with the surrounding normal tissue. Immunohistochemical staining revealed that Aggrus protein expression was detected in 10 of 11 seminomas (90.9%), but its expression was not observed in embryonal carcinomas (0/4; 0%). Specific markers for seminomas have not been reported, and Aggrus is a potential diagnostic marker for seminomas and may be associated with malignancies of the testis. PMID- 15361851 TI - Aberrations of the Chk2 tumour suppressor in advanced urinary bladder cancer. AB - Checkpoint kinase 2 (Chk2) is a tumour suppressor and signal transducer in genome integrity checkpoints that coordinate cell-cycle progression with DNA repair or cell death in response to DNA damage. Defects of Chk2 occur in subsets of diverse sporadic malignancies and predispose to several types of hereditary carcinomas. However, the status of Chk2 in tumours of the urinary bladder remains unknown. Here, we report that among 58 advanced (grade T2-T4) human bladder carcinomas, immunohistochemical analysis revealed tumour-specific reduction or lack of Chk2 protein in 6 (10.3%) cases. Genetic analysis of the latter subset showed that a Chk2-negative carcinoma #668 harboured a truncating mutation 1100delC, in one Chk2 allele and loss of the corresponding second allele. The 1100delC mutation was also found in the germ line of this patient. Sequencing of TP53 in tumour #668 identified two missense mutations. Furthermore, the vast majority of the tumours showed 'unscheduled' activatory phosphorylation on Thr68 of Chk2 in the absence of any DNA-damaging treatment. Our results indicate that the otherwise dormant DNA damage signal transducer Chk2 is aberrantly and constitutively activated in invasive urinary bladder carcinomas, and that such likely proapoptotic checkpoint signalling can be disabled by inactivation of Chk2 and/or p53 tumour suppressors in subsets of these tumours. PMID- 15361852 TI - Epstein-Barr virus (EBV) LMP2A mediates B-lymphocyte survival through constitutive activation of the Ras/PI3K/Akt pathway. AB - Epstein-Barr virus (EBV) establishes a lifelong latent infection in host B cells and is associated with the development of a variety of malignancies. The viral LMP2A protein mediates viral latency by mimicking a constitutively activated B cell receptor (BCR). In vivo LMP2A provides developmental and survival signals to BCR-negative B cells, allowing them to survive in peripheral lymphoid organs. In this study, we have demonstrated that Ras is constitutively active in peripheral, BCR-negative B cells from LMP2A transgenic mice. Furthermore, increased expression of activated Ras correlated with elevated levels of Bcl-xL expression and a slower migrating, band-shifted form of Bcl-2. B cells from LMP2A transgenic mice were sensitive to apoptosis induction in the presence of specific inhibitors of Ras, phosphatidylinositol 3-kinase (PI3K), and Akt, indicating that LMP2A activates the Ras/PI3K/Akt pathway to mediate B-cell survival. Increased B-cell apoptosis correlated with reduced expression of Bcl-xL, suggesting that this Bcl 2 family member may be involved in apoptosis inhibition mediated by LMP2A. The ability of LMP2A to activate constitutively the Ras pathway, a common event during tumorigenesis, suggests that this viral protein plays an active role in the development of EBV-associated malignancies. PMID- 15361853 TI - Alternative splicing and mutation status of CHEK2 in stage III breast cancer. AB - The DNA damage checkpoint kinase, CHK2, promotes growth arrest or apoptosis through phosphorylating targets such as Cdc25A, Cdc25C, BRCA1, and p53. Both germline and somatic loss-of-function CHEK2 mutations occur in human tumours, the former linked to the Li-Fraumeni syndrome, and the latter found in diverse types of sporadic malignancies. Here we examined the status of CHK2 by genetic and immunohistochemical analyses in 53 breast carcinomas previously characterized for TP53 status. We identified two CHEK2 mutants, 470T>C (Ile157Thr), and a novel mutation, 1368insA leading to a premature stop codon in exon 13. The truncated protein encoded by CHEK2 carrying the 1368insA was stable yet mislocalized to the cytoplasm in tumour sections and when ectopically expressed in cultured cells. Unexpectedly, we found CHEK2 to be subject to extensive alternative splicing, with some 90 splice variants detected in our tumour series. While all cancers expressed normal-length CHEK2 mRNA together with the spliced transcripts, we demonstrate and/or predict some of these splice variants to lack CHK2 function and/or localize aberrantly. We conclude that cytoplasmic sequestration may represent a novel mechanism to disable CHK2, and propose to further explore the significance of the complex splicing patterns of this tumour suppressor gene in oncogenesis. PMID- 15361854 TI - Deacetylation of p53 after nerve growth factor treatment in PC12 cells as a post translational modification mechanism of neurotrophin-induced tumor suppressor activation. AB - The tumor suppressor protein p53 is a transcription factor that regulates the response to cellular insults such as DNA damage and growth factor withdrawal. Transcriptional activity of p53 requires post-translational modification by phosphorylation and acetylation. This study used site-specific antibodies to demonstrate that nerve growth factor (NGF) treatment of PC12 cells results in p53 deacetylation at lysine (Lys) 382. Histone deacetylase (HDAC) activity, measured by a direct fluorescent assay, was increased after NGF treatment and peaked before p53 deacetylation. Inhibition of HDAC by trichostatin blocked the deacetylation of p53 and its transcriptional activity toward a reporter gene construct. Comparison of PC12 with PC12 cells containing a temperature-sensitive, dominant-negative construct showed that p53 deacetylation required functional p53. Inhibitors of MAP kinase that block p53 transactivation and inhibitors of TrkA receptor also abolished HDAC activation, indicating that deacetylation of p53 is an NGF-dependent post-translational mechanism of p53 activation. Finally, NGF or serum withdrawal did not lead to p53 deacetylation. A model is proposed in which the acetylation status of Lys 382 of p53 discriminates between cell cycle arrest and apoptosis. PMID- 15361855 TI - Whole genome expression profiling of advance stage papillary serous ovarian cancer reveals activated pathways. AB - Ovarian cancer is the most lethal type of gynecologic cancer in the Western world. The high case fatality rate is due in part because most ovarian cancer patients present with advanced stage disease which is essentially incurable. In order to obtain a whole genome assessment of aberrant gene expression in advanced ovarian cancer, we used oligonucleotide microarrays comprising over 40,000 features to profile 37 advanced stage papillary serous primary carcinomas. We identified 1191 genes that were significantly (P < 0.001) differentially regulated between the ovarian cancer specimens and normal ovarian surface epithelium. The microarray data were validated using real time RT-PCR on 14 randomly selected differentially regulated genes. The list of differentially expressed genes includes ones that are involved in cell growth, differentiation, adhesion, apoptosis and migration. In addition, numerous genes whose function remains to be elucidated were also identified. The microarray data were imported into PathwayAssist software to identify signaling pathways involved in ovarian cancer tumorigenesis. Based on our expression results, a signaling pathway associated with tumor cell migration, spread and invasion was identified as being activated in advanced ovarian cancer. The data generated in this study represent a comprehensive list of genes aberrantly expressed in serous papillary ovarian adenocarcinoma and may be useful for the identification of potentially new and novel markers and therapeutic targets for ovarian cancer. PMID- 15361856 TI - Intron-encoded homing endonuclease I-TevI also functions as a transcriptional autorepressor. AB - Customary binding sites of intron-encoded homing endonucleases lie within cognate intronless alleles, at the so-called homing sites. Here, we describe a novel, high-affinity binding site for I-TevI endonuclease, encoded within the group I td intron of phage T4. This site is an operator that overlaps the T4 late promoter, which drives I-TevI expression from within the td intron. I-TevI binds the operator and homing sites with equal affinity, and functions as a transcriptional autorepressor. Distinct sequence and spacing requirements of the catalytic domain result in reduced cleavage activity on operator DNA. Crystallographic studies showed that the overall interactions of the DNA-binding domain with the operator and homing sites are similar, but have some different hydrogen-bonding contacts. We present a model in which the flexibility in protein-DNA interactions allows I TevI to bind variant intronless alleles to promote intron mobility while facilitating its function in autorepression, and thereby persistence in its host. PMID- 15361857 TI - eIF4G is required for the pioneer round of translation in mammalian cells. AB - Nonsense-mediated mRNA decay (NMD) in mammalian cells targets cap-binding protein 80 (CBP80)-bound mRNA during or after a pioneer round of translation. It is unknown whether eukaryotic translation initiation factor 4G (eIF4G) functions in the pioneer round. We show that baculovirus-produced CBP80 and CBP20 independently interact with eIF4GI. The interactions between eIF4G and the heterodimer CBP80/20 suggest that eIF4G has a function in the pioneer initiation complex rather than merely a presence during remodeling to the steady-state complex. First, NMD is inhibited upon eIF4G cleavage by HIV-2 or poliovirus 2A protease. Second, eIF4GI coimmunopurifies with pre-mRNA, indicating that it associates with transcripts before the pioneer round. Third, eIF4G immunopurifies with Upf NMD factors and eIF4AIII, which are constituents of the pioneer translation initiation complex. We propose a model in which eIF4G serves to connect CBP80/20 with other initiation factors during the pioneer round of translation. PMID- 15361858 TI - Control of human potassium channel inactivation by editing of a small mRNA hairpin. AB - Genomic recoding by A-->I RNA editing plays an important role in diversifying the proteins involved in electrical excitability. Here, we describe editing of an intronless potassium channel gene. A small region of human K(V)1.1 mRNA sequence directs efficient modification of one adenosine by human adenosine deaminase acting on RNA 2 (hADAR2). Mutational analysis shows that this region adopts a hairpin structure. Electrophysiological characterization reveals that the editing event (I/V) profoundly affects channel inactivation conferred by accessory beta subunits. Drosophila melanogaster Shaker channels, mimicking this editing event through mutation, exhibit a similar effect. In addition, we demonstrate that mRNAs for the paralogous D. melanogaster Shab potassium channel are edited at the same position by fly ADAR-a clear example of convergent evolution driven by adenosine deamination. These results suggest an ancient and key regulatory role for this residue in K(V) channels. PMID- 15361860 TI - Octaheme tetrathionate reductase is a respiratory enzyme with novel heme ligation. AB - We have isolated a soluble cytochrome from Shewanella oneidensis that contains eight covalently attached heme groups and determined its crystal structure. One of these hemes exhibits novel ligation of the iron atom by the epsilon-amino group of a lysine residue, despite its attachment via a typical CXXCH motif. This heme is most likely the active site for tetrathionate reduction, a reaction catalyzed efficiently by this enzyme. PMID- 15361859 TI - A unique E1-E2 interaction required for optimal conjugation of the ubiquitin-like protein NEDD8. AB - Ubiquitin-like proteins (UBLs) such as NEDD8 are transferred to their targets by distinct, parallel, multienzyme cascades that involve the sequential action of E1, E2 and E3 enzymes. How do enzymes within a particular UBL conjugation cascade interact with each other? We report here that the unique N-terminal sequence of NEDD8's E2, Ubc12, selectively recruits NEDD8's E1 to promote thioester formation between Ubc12 and NEDD8. A peptide corresponding to Ubc12's N terminus (Ubc12N26) specifically binds and inhibits NEDD8's E1, the heterodimeric APPBP1-UBA3 complex. The structure of APPBP1-UBA3- Ubc12N26 reveals conserved Ubc12 residues docking in a groove generated by loops conserved in UBA3s but not other E1s. These data explain why the Ubc12-UBA3 interaction is unique to the NEDD8 pathway. These studies define a novel mechanism for E1-E2 interaction and show how enzymes within a particular UBL conjugation cascade can be tethered together by unique protein-protein interactions emanating from their common structural scaffolds. PMID- 15361861 TI - Structure of the periplasmic chaperone Skp suggests functional similarity with cytosolic chaperones despite differing architecture. AB - The 17-kDa protein (Skp) of Escherichia coli is a homotrimeric periplasmic chaperone for newly synthesized outer-membrane proteins. Here we present its X ray structure at a resolution of 2.35 A. Three hairpin-shaped alpha-helical extensions reach out by approximately 60 A from a trimerization domain, which is composed of three intersubunit beta-sheets that wind around a central axis. The alpha-helical extensions approach each other at their distal turns, resulting in a fold that resembles a 'three-pronged grasping forceps'. The overall shape of Skp is reminiscent of the cytosolic chaperone prefoldin, although it is based on a radically different topology. The peculiar architecture, with apparent plasticity of the prongs and distinct electrostatic and hydrophobic surface properties, supports the recently proposed biochemical mechanism of this chaperone: formation of a Skp(3)-Omp complex protects the outer membrane protein from aggregation during passage through the bacterial periplasm. PMID- 15361862 TI - The nisin-lipid II complex reveals a pyrophosphate cage that provides a blueprint for novel antibiotics. AB - The emerging antibiotics-resistance problem has underlined the urgent need for novel antimicrobial agents. Lantibiotics (lanthionine-containing antibiotics) are promising candidates to alleviate this problem. Nisin, a member of this family, has a unique pore-forming activity against bacteria. It binds to lipid II, the essential precursor of cell wall synthesis. As a result, the membrane permeabilization activity of nisin is increased by three orders of magnitude. Here we report the solution structure of the complex of nisin and lipid II. The structure shows a novel lipid II-binding motif in which the pyrophosphate moiety of lipid II is primarily coordinated by the N-terminal backbone amides of nisin via intermolecular hydrogen bonds. This cage structure provides a rationale for the conservation of the lanthionine rings among several lipid II-binding lantibiotics. The structure of the pyrophosphate cage offers a template for structure-based design of novel antibiotics. PMID- 15361863 TI - The superhelical TPR-repeat domain of O-linked GlcNAc transferase exhibits structural similarities to importin alpha. AB - Addition of N-acetylglucosamine (GlcNAc) is a ubiquitous form of intracellular glycosylation catalyzed by the conserved O-linked GlcNAc transferase (OGT). OGT contains an N-terminal domain of tetratricopeptide (TPR) repeats that mediates the recognition of a broad range of target proteins. Components of the nuclear pore complex are major OGT targets, as OGT depletion by RNA interference (RNAi) results in the loss of GlcNAc modification at the nuclear envelope. To gain insight into the mechanism of target recognition, we solved the crystal structure of the homodimeric TPR domain of human OGT, which contains 11.5 TPR repeats. The repeats form an elongated superhelix. The concave surface of the superhelix is lined by absolutely conserved asparagines, in a manner reminiscent of the peptide binding site of importin alpha. Based on this structural similarity, we propose that OGT uses an analogous molecular mechanism to recognize its targets. PMID- 15361864 TI - Volumetric computed tomography (VCT): a new technology for noninvasive, high resolution monitoring of tumor angiogenesis. AB - Volumetric computed tomography (VCT) is a technology in which area detectors are used for imaging large volumes of a subject with isotropic imaging resolution. We are experimenting with a prototype VCT scanner that uses flat-panel X-ray detectors and is designed for high-resolution three-dimensional (3D) imaging. Using this technique, we have demonstrated microangiography of xeno-transplanted skin squamous cell carcinomas in nude mice. VCT shows the vessel architecture of tumors and animals with greater detail and plasticity than has previously been achieved, and is superior to contrast-enhanced magnetic resonance (MR) angiography. VCT and MR images correlate well for larger tumor vessels, which are tracked from their origin on 3D reconstructions of VCT images. When compared with histology, small tumor vessels with a diameter as small as 50 microm were clearly visualized. Furthermore, imaging small vessel networks inside the tumor tissue improved discrimination of vital and necrotic regions. Thus, VCT substantially improves imaging of vascularization in tumors and offers a promising tool for preclinical studies of tumor angiogenesis and antiangiogenic therapies. PMID- 15361866 TI - Crucial functions of the Rap1 effector molecule RAPL in lymphocyte and dendritic cell trafficking. AB - Immunosurveillance requires the coordinated regulation of chemokines and adhesion molecules to guide immune cell migration. However, the critical molecule for governing the high trafficking capability of immune cells is not clear. Here we show that the effector molecule RAPL is indispensable in the integrin-mediated adhesion and migration of lymphocytes and dendritic cells. RAPL deficiency caused defective chemokine-triggered lymphocyte adhesion and migration to secondary lymphoid organs, resulting in atrophic lymphoid follicles and deficient marginal zone B cells, concomitant with increased immature B cells in the blood. Furthermore, splenic dendritic cells were diminished and defective in adhesion. After being activated with inflammatory stimuli, skin and splenic dendritic cells failed to migrate into either the draining lymph nodes or the white pulp of the spleen. Thus, RAPL is a crucial immune cell trafficking regulator essential for immunosurveillance. PMID- 15361865 TI - Inhaled nebulized nitrite is a hypoxia-sensitive NO-dependent selective pulmonary vasodilator. AB - The blood anion nitrite contributes to hypoxic vasodilation through a heme-based, nitric oxide (NO)-generating reaction with deoxyhemoglobin and potentially other heme proteins. We hypothesized that this biochemical reaction could be harnessed for the treatment of neonatal pulmonary hypertension, an NO-deficient state characterized by pulmonary vasoconstriction, right-to-left shunt pathophysiology and systemic hypoxemia. To test this, we delivered inhaled sodium nitrite by aerosol to newborn lambs with hypoxic and normoxic pulmonary hypertension. Inhaled nitrite elicited a rapid and sustained reduction ( approximately 65%) in hypoxia-induced pulmonary hypertension, with a magnitude approaching that of the effects of 20 p.p.m. NO gas inhalation. This reduction was associated with the immediate appearance of NO in expiratory gas. Pulmonary vasodilation elicited by aerosolized nitrite was deoxyhemoglobin- and pH-dependent and was associated with increased blood levels of iron-nitrosyl-hemoglobin. Notably, from a therapeutic standpoint, short-term delivery of nitrite dissolved in saline through nebulization produced selective, sustained pulmonary vasodilation with no clinically significant increase in blood methemoglobin levels. These data support the concept that nitrite is a vasodilator acting through conversion to NO, a process coupled to hemoglobin deoxygenation and protonation, and evince a new, simple and inexpensive potential therapy for neonatal pulmonary hypertension. PMID- 15361867 TI - GA binding protein regulates interleukin 7 receptor alpha-chain gene expression in T cells. AB - The interleukin 7 receptor alpha-chain (IL-7Ralpha) is essential for T cell development in both humans and mice and for B cell development in mice. Whereas the transcription factor PU.1 regulates IL-7Ralpha expression in mouse pro-B cells via a GGAA motif, we demonstrate here that GA binding protein (GABP) bound to this site and was essential in the regulation of IL-7Ralpha expression in T cells, where PU.1 is not expressed. Moreover, IL-7Ralpha expression was diminished substantially in thymocytes but was normal on B220(+) fetal liver cells from mouse embryos with diminished expression of GABPalpha. Thus, GABP is essential for the regulation of IL-7Ralpha expression in T cells, and the differential regulation of IL-7Ralpha in distinct lymphoid lineages is achieved at least in part by differential recruitment of factors to the same GGAA motif. PMID- 15361868 TI - Interferon-alpha induction through Toll-like receptors involves a direct interaction of IRF7 with MyD88 and TRAF6. AB - Toll-like receptors (TLRs) are involved in the recognition of microbial pathogens. A subset of TLRs, TLR7, TLR8 and TLR9, induces antiviral responses by producing interferon-alpha (IFN-alpha). Production of IFN-alpha is dependent on the Toll-interleukin-1 receptor domain-containing adaptor MyD88. Here we show that MyD88 formed a complex with the transcription factor IRF7 but not with IRF3. The death domain of MyD88 interacted with an inhibitory domain of IRF7, and this interaction resulted in activation of the IFN-alpha-dependent promoters. Furthermore, the adaptor molecule TRAF6 also bound and activated IRF7. Ubiquitin ligase activity of TRAF6 was required for IRF7 activation. These results indicate that TLR-mediated IFN-alpha induction requires the formation of a complex consisting of MyD88, TRAF6 and IRF7 as well as TRAF6-dependent ubiquitination. PMID- 15361869 TI - Early B cell factor cooperates with Runx1 and mediates epigenetic changes associated with mb-1 transcription. AB - Cd79a (called mb-1 here) encodes the Ig-alpha signaling component of the B cell receptor. The early B cell-specific mb-1 promoter was hypermethylated at CpG dinucleotides in hematopoietic stem cells but became progressively unmethylated as B cell development proceeded. The transcription factor Pax5 activated endogenous mb-1 transcription in a plasmacytoma cell line, but could not when the promoter was methylated. In this context, early B cell factor (EBF), a transcription factor required for B lymphopoiesis, potentiated activation of mb-1 by Pax5. EBF and the basic helix-loop-helix transcription factor E47 each contributed to epigenetic modifications of the mb-1 promoter, including CpG demethylation and nucleosomal remodeling. EBF function was enhanced by interaction with the transcription factor Runx1. These data suggest a molecular basis for the hierarchical dependence of Pax5 function on EBF and E2A in B lymphocyte development. PMID- 15361870 TI - A specific requirement for PDGF-C in palate formation and PDGFR-alpha signaling. AB - PDGF-C is a member of the platelet-derived growth factor (PDGF) family, which signals through PDGF receptor (PDGFR) alphaalpha and alphabeta dimers. Here we show that Pdgfc(-/-) mice die in the perinatal period owing to feeding and respiratory difficulties associated with a complete cleft of the secondary palate. This phenotype was less severe than that of Pdgfra(-/-) embryos. Pdgfc(-/ ) Pdgfa(-/-) embryos developed a cleft face, subepidermal blistering, deficiency of renal cortex mesenchyme, spina bifida and skeletal and vascular defects. Complete loss of function of both ligands, therefore, phenocopied the loss of PDGFR-alpha function, suggesting that both PDGF-A and PDGF-C signal through PDGFR alpha to regulate the development of craniofacial structures, the neural tube and mesodermal organs. Our results also show that PDGF-C signaling is a new pathway in palatogenesis, different from, and independent of, those previously implicated. PMID- 15361871 TI - MicroRNA-responsive 'sensor' transgenes uncover Hox-like and other developmentally regulated patterns of vertebrate microRNA expression. AB - MicroRNAs (miRNAs) are a class of short ( approximately 22-nt) noncoding RNA molecules that downregulate expression of their mRNA targets. Since their discovery as regulators of developmental timing in Caenorhabditis elegans, hundreds of miRNAs have been identified in both animals and plants. Here, we report a technique for visualizing detailed miRNA expression patterns in mouse embryos. We elucidate the tissue-specific expression of several miRNAs during embryogenesis, including two encoded by genes embedded in homeobox (Hox) clusters, miR-10a and miR-196a. These two miRNAs are expressed in patterns that are markedly reminiscent of those of Hox genes. Furthermore, miR-196a negatively regulates Hoxb8, indicating that its restricted expression pattern probably reflects a role in the patterning function of the Hox complex. PMID- 15361872 TI - Active genes dynamically colocalize to shared sites of ongoing transcription. AB - The intranuclear position of many genes has been correlated with their activity state, suggesting that migration to functional subcompartments may influence gene expression. Indeed, nascent RNA production and RNA polymerase II seem to be localized into discrete foci or 'transcription factories'. Current estimates from cultured cells indicate that multiple genes could occupy the same factory, although this has not yet been observed. Here we show that, during transcription in vivo, distal genes colocalize to the same transcription factory at high frequencies. Active genes are dynamically organized into shared nuclear subcompartments, and movement into or out of these factories results in activation or abatement of transcription. Thus, rather than recruiting and assembling transcription complexes, active genes migrate to preassembled transcription sites. PMID- 15361873 TI - The EBV lytic switch protein, Z, preferentially binds to and activates the methylated viral genome. AB - DNA methylation promotes gene silencing, yet the Epstein-Barr virus immediate early protein, BZLF1 (Z), converts the virus from the latent to the lytic form of infection even when the viral genome is highly methylated. Here we show that methylation of CpG motifs in Z-responsive elements of the viral BRLF1 immediate early promoter enhances Z binding to, and activation of, this promoter. Demethylation of the viral genome impairs Z activation of lytic viral genes. Z is the first transcription factor that preferentially binds to, and activates, a methylated promoter. These results identify an unexpected mechanism by which Epstein-Barr virus circumvents the inhibitory effects of viral genome methylation. PMID- 15361874 TI - Fusion of NUP214 to ABL1 on amplified episomes in T-cell acute lymphoblastic leukemia. AB - In T-cell acute lymphoblastic leukemia (T-ALL), transcription factors are known to be deregulated by chromosomal translocations, but mutations in protein tyrosine kinases have only rarely been identified. Here we describe the extrachromosomal (episomal) amplification of ABL1 in 5 of 90 (5.6%) individuals with T-ALL, an aberration that is not detectable by conventional cytogenetics. Molecular analyses delineated the amplicon as a 500-kb region from chromosome band 9q34, containing the oncogenes ABL1 and NUP214 (refs. 5,6). We identified a previously undescribed mechanism for activation of tyrosine kinases in cancer: the formation of episomes resulting in a fusion between NUP214 and ABL1. We detected the NUP214-ABL1 transcript in five individuals with the ABL1 amplification, in 5 of 85 (5.8%) additional individuals with T-ALL and in 3 of 22 T-ALL cell lines. The constitutively phosphorylated tyrosine kinase NUP214-ABL1 is sensitive to the tyrosine kinase inhibitor imatinib. The recurrent cryptic NUP214-ABL1 rearrangement is associated with increased HOX expression and deletion of CDKN2A, consistent with a multistep pathogenesis of T-ALL. NUP214 ABL1 expression defines a new subgroup of individuals with T-ALL who could benefit from treatment with imatinib. PMID- 15361875 TI - Poly(ADP-ribosyl)ation regulates CTCF-dependent chromatin insulation. AB - Chromatin insulators demarcate expression domains by blocking the cis effects of enhancers or silencers in a position-dependent manner. We show that the chromatin insulator protein CTCF carries a post-translational modification: poly(ADP ribosyl)ation. Chromatin immunoprecipitation analysis showed that a poly(ADP ribosyl)ation mark, which exclusively segregates with the maternal allele of the insulator domain in the H19 imprinting control region, requires the bases that are essential for interaction with CTCF. Chromatin immunoprecipitation-on-chip analysis documented that the link between CTCF and poly(ADP-ribosyl)ation extended to more than 140 mouse CTCF target sites. An insulator trap assay showed that the insulator function of most of these CTCF target sites is sensitive to 3 aminobenzamide, an inhibitor of poly(ADP-ribose) polymerase activity. We suggest that poly(ADP-ribosyl)ation imparts chromatin insulator properties to CTCF at both imprinted and nonimprinted loci, which has implications for the regulation of expression domains and their demise in pathological lesions. PMID- 15361876 TI - Rapid and persistent modulation of actin dynamics regulates postsynaptic reorganization underlying bidirectional plasticity. AB - The synapse is a highly organized cellular specialization whose structure and composition are reorganized, both positively and negatively, depending on the strength of input signals. The mechanisms orchestrating these changes are not well understood. A plausible locus for the reorganization of synapse components and structure is actin, because it serves as both cytoskeleton and scaffold for synapses and exists in a dynamic equilibrium between F-actin and G-actin that is modulated bidirectionally by cellular signaling. Using a new FRET-based imaging technique to monitor F-actin/G-actin equilibrium, we show here that tetanic stimulation causes a rapid, persistent shift of actin equilibrium toward F-actin in the dendritic spines of rat hippocampal neurons. This enlarges the spines and increases postsynaptic binding capacity. In contrast, prolonged low-frequency stimulation shifts the equilibrium toward G-actin, resulting in a loss of postsynaptic actin and of structure. This bidirectional regulation of actin is actively involved in protein assembly and disassembly and provides a substrate for bidirectional synaptic plasticity. PMID- 15361877 TI - Experience can change the 'light-from-above' prior. AB - To interpret complex and ambiguous input, the human visual system uses prior knowledge or assumptions about the world. We show that the 'light-from-above' prior, used to extract information about shape from shading is modified in response to active experience with the scene. The resultant adaptation is not specific to the learned scene but generalizes to a different task, demonstrating that priors are constantly adapted by interactive experience with the environment. PMID- 15361879 TI - Human orbitofrontal cortex mediates extinction learning while accessing conditioned representations of value. AB - In extinction, an animal learns that a previously conditioned stimulus (CS+) no longer predicts delivery of a salient reinforcer (unconditioned stimulus, UCS). Rodent studies indicate that extinction relies on amygdala-prefrontal interactions and involves formation of memories that inhibit, without actually erasing, the original conditioning trace. Whether extinction learning in humans follows similar neurobiological principles is unknown. We used functional magnetic resonance imaging to measure human brain activity evoked during olfactory aversive conditioning and extinction learning. Neural responses in orbitofrontal cortex and amygdala were preferentially enhanced during extinction, suggesting potential cross-species preservation of learning mechanisms that oppose conditioning. Moreover, by manipulating UCS aversiveness via reinforcer inflation, we showed that a CS+ retains access to representations of UCS value in distinct regions of ventral prefrontal cortex, even as extinction proceeds. PMID- 15361878 TI - Rolling blackout, a newly identified PIP2-DAG pathway lipase required for Drosophila phototransduction. AB - The rolling blackout (rbo) gene encodes an integral plasma membrane lipase required for Drosophila phototransduction. Photoreceptors are enriched for the RBO protein, and temperature-sensitive rbo mutants show reversible elimination of phototransduction within minutes, demonstrating an acute requirement for the protein. The block is activity dependent, indicating that the action of RBO is use dependent. Conditional rbo mutants show activity-dependent depletion of diacylglycerol and concomitant accumulation of phosphatidylinositol phosphate and phosphatidylinositol 4,5-bisphosphate within minutes of induction, suggesting rapid downregulation of phospholipase C (PLC) activity. The RBO requirement identifies an essential regulatory step in G-protein-coupled, PLC-dependent inositol lipid signaling mediating activation of TRP and TRPL channels during phototransduction. PMID- 15361880 TI - Early and rapid perceptual learning. AB - Major, rapid performance improvements in perceptual training are often dismissed as 'task' or 'procedural' learning because they are fast and generalize within a task. We assessed the contributions of perceptual and procedural learning to improvement in an auditory tone frequency learning task in humans and found that perceptual learning accounted for between 76% and 98% of the rapid early performance improvement. PMID- 15361881 TI - Protein sequencing by mass analysis of polypeptide ladders after controlled protein hydrolysis. AB - The characterization of protein modifications is essential for the study of protein function using functional genomic and proteomic approaches. However, current techniques are not efficient in determining protein modifications. We report an approach for sequencing proteins and determining modifications with high speed, sensitivity and specificity. We discovered that a protein could be readily acid-hydrolyzed within 1 min by exposure to microwave irradiation to form, predominantly, two series of polypeptide ladders containing either the N- or C-terminal amino acid of the protein, respectively. Mass spectrometric analysis of the hydrolysate produced a simple mass spectrum consisting of peaks exclusively from these polypeptide ladders, allowing direct reading of amino acid sequence and modifications of the protein. As examples, we applied this technique to determine protein phosphorylation sites as well as the sequences and several previously unknown modifications of 28 small proteins isolated from Escherichia coli K12 cells. This technique can potentially be automated for large-scale protein annotation. PMID- 15361882 TI - Prediction of sequence-dependent and mutational effects on the aggregation of peptides and proteins. AB - We have developed a statistical mechanics algorithm, TANGO, to predict protein aggregation. TANGO is based on the physico-chemical principles of beta-sheet formation, extended by the assumption that the core regions of an aggregate are fully buried. Our algorithm accurately predicts the aggregation of a data set of 179 peptides compiled from the literature as well as of a new set of 71 peptides derived from human disease-related proteins, including prion protein, lysozyme and beta2-microglobulin. TANGO also correctly predicts pathogenic as well as protective mutations of the Alzheimer beta-peptide, human lysozyme and transthyretin, and discriminates between beta-sheet propensity and aggregation. Our results confirm the model of intermolecular beta-sheet formation as a widespread underlying mechanism of protein aggregation. Furthermore, the algorithm opens the door to a fully automated, sequence-based design strategy to improve the aggregation properties of proteins of scientific or industrial interest. PMID- 15361883 TI - Regulation of B-cell survival by BAFF-dependent PKCdelta-mediated nuclear signalling. AB - Approximately 65% of B cells generated in human bone marrow are potentially harmful autoreactive B cells. Most of these cells are clonally deleted in the bone marrow, while those autoreactive B cells that escape to the periphery are anergized or perish before becoming mature B cells. Escape of self-reactive B cells from tolerance permits production of pathogenic auto-antibodies; recent studies suggest that extended B lymphocyte survival is a cause of autoimmune disease in mice and humans. Here we report a mechanism for the regulation of peripheral B-cell survival by serine/threonine protein kinase Cdelta (PKCdelta): spontaneous death of resting B cells is regulated by nuclear localization of PKCdelta that contributes to phosphorylation of histone H2B at serine 14 (S14 H2B). We show that treatment of B cells with the potent B-cell survival factor BAFF ('B-cell-activating factor belonging to the TNF family') prevents nuclear accumulation of PKCdelta. Our data suggest the existence of a previously unknown BAFF-induced and PKCdelta-mediated nuclear signalling pathway which regulates B cell survival. PMID- 15361884 TI - p19ARF directly and differentially controls the functions of c-Myc independently of p53. AB - Increased expression of the oncogenic transcription factor c-Myc causes unregulated cell cycle progression. c-Myc can also cause apoptosis, but it is not known whether the activation and/or repression of c-Myc target genes mediates these diverse functions of c-Myc. Because unchecked cell cycle progression leads to hyperproliferation and tumorigenesis, it is essential for tumour suppressors, such as p53 and p19ARF (ARF), to curb cell cycle progression in response to increased c-Myc (refs 2, 3). Increased c-Myc has previously been shown to induce ARF expression, which leads to cell cycle arrest or apoptosis through the activation of p53 (ref. 4). Here we show that ARF can inhibit c-Myc by a unique and direct mechanism that is independent of p53. When c-Myc increases, ARF binds with c-Myc and dramatically blocks c-Myc's ability to activate transcription and induce hyperproliferation and transformation. In contrast, c-Myc's ability to repress transcription is unaffected by ARF and c-Myc-mediated apoptosis is enhanced. These differential effects of ARF on c-Myc function suggest that separate molecular mechanisms mediate c-Myc-induced hyperproliferation and apoptosis. This direct feedback mechanism represents a p53-independent checkpoint to prevent c-Myc-mediated tumorigenesis. PMID- 15361885 TI - Hedgehog signalling in prostate regeneration, neoplasia and metastasis. AB - Metastatic cancers adopt certain properties of normal cells in developing or regenerating organs, such as the ability to proliferate and alter tissue organization. We find here that activity of the Hedgehog (Hh) signalling pathway, which has essential roles in developmental patterning, is required for regeneration of prostate epithelium, and that continuous pathway activation transforms prostate progenitor cells and renders them tumorigenic. Elevated pathway activity furthermore distinguishes metastatic from localized prostate cancer, and pathway manipulation can modulate invasiveness and metastasis. Pathway activity is triggered in response to endogenous expression of Hh ligands, and is dependent upon the expression of Smoothened, an essential Hh response component that is not expressed in benign prostate epithelial cells. Monitoring and manipulating Hh pathway activity may thus offer significant improvements in diagnosis and treatment of prostate cancers with metastatic potential. PMID- 15361886 TI - Disparate systemic and renal blocking properties of angiotensin II antagonists during exogenous angiotensin II administration: implications for treatment. AB - Generation of angiotensin II (Ang II) contributes to the pathogenesis of cardiovascular diseases. Owing to the existence of high levels of Ang II within the kidney, blockade of the intrarenal Ang II levels may be important since long term outcome seems not only to be related to blood pressure per se. This was a prospective, randomized, double-blind, placebo-controlled study, with crossover design. We examined in 13 patients with mild to moderate hypertension the specific systemic and renal blocking properties of two different Ang II receptor blockers during a wide range of Ang II concentrations, 24 h post dose. The effects were evaluated after 4 weeks treatment with candesartan cilexetil (16 mg OD), losartan (50 mg OD) and placebo using clearance techniques. Candesartan reduced the 24 h blood pressure better than losartan (138(*)/87+/-12/8 vs 145/89+/-12/7 mmHg, (*)P<0.05 vs losartan) and placebo. Despite the lower blood pressure, candesartan attenuated the Ang II-induced response on ERPF and RVR markedly better than losartan or placebo. The GFR decreased, as expected, with placebo, but remained stable with candesartan. The present study demonstrates that in hypertensive patients candesartan and to a lesser degree losartan are effective in blocking the systemic and renal effects of Ang II during a wide range of Ang II infusion rates. Interestingly, 24 h post dose, candesartan effectively diminished the change in ERPF as well as GFR. This sustained renal effect of candesartan may be of importance, especially in pathophysiological circumstances in which (high renal levels of) Ang II contributes to cardiovascular damage. PMID- 15361887 TI - Racial/ethnic differences in prehypertension in American adults: population and relative attributable risks of abdominal obesity. AB - To estimate the risk and population attributable risk of prehypertension that is due to abdominal obesity in White, Black and Hispanic American adults. To determine how much of the relative difference in the risk of prehypertension between high-risk Blacks and Hispanics and the low-risk group Whites that is attributable to their differences in abdominal obesity. Data (n=4016) from the 1999 to 2000 US National Health and Nutrition Examination Surveys were used in this study. Abdominal obesity was defined as waist circumference >/=102 and >/=88 cm in men and women, respectively. Prehypertension was defined as not being on antihypertensive medication and having systolic blood pressure of 120-139 mmHg or diastolic blood pressure of 80-89 mmHg. Odds ratio from the logistic regression analysis was used to estimate the risk of prehypertension that was due to abdominal obesity. To estimate prehypertension risk differences between low-risk Whites and high-risk Blacks and Hispanics that was due to abdominal obesity, we estimated relative attributable risk. Statistical adjustments were made for age, blood glucose, total cholesterol, current smoking and exercise. Abdominal obesity was associated with increased odds of prehypertension in Whites, Blacks and Hispanics. In men, abdominal obesity was associated with 44, 90 and 98% increased odds of prehypertension in Whites, Blacks and Hispanics, respectively. The corresponding values in women were 112, 198 and 104%. Proportions of risk of prehypertension explained by abdominal obesity were 15.2, 22 and 25.8% in White men, Black men and Hispanic men, respectively. The corresponding values in women were 38.8, 58.6 and 32.5%. Approximately, 7% of the differences in the risk of developing prehypertension between White and Black men and between White and Hispanic men may be attributable to differences in rates of abdominal obesity. The analogous values for women were approximately 39.7 and approximately 16.5%, respectively. In conclusion, despite having lower rates of abdominal obesity than their counterparts, Black men, Hispanic men and Hispanic women had high population attributable risks, indicating that factors other than abdominal obesity may have important explanatory power for racial differences in prehypertension in these groups. However, in Black women reduction in risk of prehypertension could be possible by instituting public health measures to reduce abdominal obesity to the levels seen in White women. Intervention programmes designed to reduce overall obesity may also lead to reduction of abdominal obesity, and consequently may curb prehypertension in these population groups. Life-style modification, including diet and exercise, may have public health significance in reducing the incidence of prehypertension in these populations. PMID- 15361888 TI - Comparing office-based and ambulatory blood pressure monitoring in clinical trials. AB - Ambulatory blood pressure monitoring (ABPM) is commonly used in clinical trials. Yet, its ability to detect blood pressure (BP) change in comparison to multiple office-based measurements has received limited attention. We recorded ambulatory and five daily pairs of random zero (RZ) BPs pre- and post-intervention on 321 adult participants in the multicentre Dietary Approaches to Stop Hypertension trial. Treatment effect estimates measured by ambulatory monitoring were similar to those measured by RZ and did not differ significantly for waking vs 24-h ambulatory measurements. For systolic BP, the standard deviations of change in mean 24-h ambulatory BP (8.0 mmHg among hypertensives and 6.0 mmHg among nonhypertensives) were comparable to or lower than the corresponding standard deviations of change in RZ-BP based on five daily readings (8.9 and 5.9 mmHg). The standard deviations of change for mean waking ambulatory BP (8.7 and 6.7 mmHg) were comparable to those obtained using three to four daily RZ readings. Results for diastolic BP were qualitatively similar. Ambulatory monitoring was more efficient (ie, a smaller sample size could detect a given BP change) than three to four sets of daily RZ readings and required fewer clinic visits. The average of 33 ambulatory BP readings during the waking hours had an efficiency comparable to that from the mean of four daily pairs of RZ-BPs. Participants readily accepted the ABPM devices, and their use requires less staff training. ABPM provides a useful alternative to RZ-BP measurements in clinical trials. PMID- 15361889 TI - Genetic variations related to hypertension: a review. AB - Hypertension is a complex multifactorial disorder with genetic, environmental and demographic factors contributing to its prevalence. The genetic element contribution to blood pressure variation ranges from 30 to 50%. Therefore, identifying hypertension susceptibility genes will help understanding the pathophysiology of the disease. In addition to the potential impact of genomic information in selecting antihypertensive drug therapy, it may also help in recognizing those at risk of developing the disease, which may lead to new preventive approaches. Several strategies and methods have been used to identify hypertension susceptibility genes. Currently, genetic analysis of such data produced complex results, which makes it difficult to draw final conclusion on the use of genomic data in management of hypertension. This review attempts to summarize present known genetic variations that may be implicated in the pathogenesis of hypertension and to discuss various research strategies used to identify them. It also highlights some of the opportunities and challenges, which may be encountered in interpreting the value of these genetic variations to improve management of hypertension. PMID- 15361890 TI - Influence of dietary sodium on the renin-angiotensin-aldosterone system and prevalence of left ventricular hypertrophy by EKG criteria. AB - We investigated the interplay of dietary sodium and renin-angiotensin-aldosterone system (RAAS) activity with the prevalence of left ventricular hypertrophy (LVH) in essential hypertension. Electrocardiograms (EKG) were reviewed for the presence of LVH in 160 hypertensive patients. We then compared the rate of LVH to levels of plasma renin activity (PRA) and serum aldosterone under high and low sodium diet conditions. On high sodium diet, serum aldosterone was significantly higher (7.7+/-0.93 vs 5.7+/-0.35 ng/dl, P=0.02) in participants with LVH. With low sodium diet and upright posture, PRA was significantly lower in subjects with LVH vs those without (5.6+/-1.1 vs 7.6+/-0.56 ng/ml/h, P=0.026). Aldosterone levels on low sodium diet were not different between those with and those without LVH. PRA was then dichotomized at the lowest quartile under low sodium/upright posture conditions to define a 'low renin' group. In a multivariate logistic regression containing renin status (low renin vs normal/high renin), aldosterone on a high sodium diet, age, body mass index, gender, race, duration of hypertension, systolic and diastolic blood pressure and salt-sensitivity only low renin status on a low sodium diet (P=0.019) and serum aldosterone on a high sodium diet (P=0.04) were significant predictors of LVH. Thus, reduced modulation of renin activity in response to sodium restriction and an increased aldosterone on a high sodium diet appear to identify characteristics of hypertensive patients predisposed to abnormal cardiac remodelling. PMID- 15361891 TI - Independent association between inflammatory markers (C-reactive protein, interleukin-6, and TNF-alpha) and essential hypertension. AB - High blood pressure (HBP) has been associated with elevated C-reactive protein (CRP), a marker of chronic mild inflammation. However, the association between HBP and other inflammatory markers, particularly interleukin 6 (IL-6) and tumour necrosis alpha (TNF-alpha), has not been evaluated in well-controlled studies. We examined the cross-sectional relationship between IL-6, TNF-alpha, and CRP and HBP in a random sample of 196 healthy subjects. All markers were measured in duplicate with high-sensitivity ELISA tests. Three blood pressure (BP) measurments were averaged for the analysis, and subjects with systolic BP >or=140 and/or diastolic BP >or=90 mmHg were considered hypertensive. Log binomial regression was used to estimate multivariate-adjusted prevalence ratios (PR) of HBP. Of the subjects, 40% (79) were hypertensive (mean age: 44 years; range 30 64). After adjustment for age, sex, body mass index, family history of HBP, and the level of the other inflammatory markers, subjects in the second (PR: 3.10, P=0.003), third (PR: 2.32; P=0.031), and fourth quartiles (PR: 2.30; P=0.036) of IL-6 were more than twice as likely to be hypertensive than those in the first quartile. Corresponding PR estimates for TNF-alpha levels were 1.41 (P=0.014) for the second; 1.59 (P=0.001) for the third; and 1.61 (P=0.025) for the fourth quartile. The CRP-HBP association was not statistically significant. Our results suggest that TNF-alpha and IL-6 could be independent risk factors for HBP in apparently healthy subjects. Nevertheless, the temporal relationship between elevated inflammation markers and HBP should be ascertained in prospective cohort studies. PMID- 15361892 TI - Long-term effects of statins on arterial pressure and stiffness of hypertensives. AB - Although lowering blood pressure (BP) reduces aortic stiffness, achieving the recommended BP goal can be difficult. Recent studies have shown that short-term use of statins can reduce BP significantly. To determine the long-term effects of statins on BP and aortic stiffness, a single-blind randomized prospective study was performed on 85 hyperlipidaemic hypertensive patients whose BP was insufficiently controlled by antihypertensive therapy. Every 3 months, aortic stiffness was assessed by measuring pulse wave velocity (PWV). Patients were randomly allocated to groups treated with pravastatin, simvastatin, fluvastatin, or a nonstatin antihyperlipidaemic drug. No significant differences in patient characteristics, kinds of antihypertensive drugs, BP, ankle brachial index, PWV, or serum lipid, creatinine, or C-reactive protein levels were found between the four groups at the start of the study. During the 12-month treatment period, PWV did not change in the pravastatin group or nonstatin group, but it was transiently reduced in the simvastatin group and significantly decreased in the fluvastatin group, even though the doses of the statins used in this study were lower than the usually prescribed dose. All four antihyperlipidaemic drugs significantly decreased serum cholesterol levels without affecting BP, ankle brachial index, or serum triglyceride levels. The C-reactive protein serum levels decreased significantly in the three statin groups but not in the nonstatin group. These results suggest that long-term use of fluvastatin by hyperlipidaemic hypertensive patients is associated with a significant reduction in aortic stiffness without any effect on BP. PMID- 15361893 TI - Trends in antihypertensive and lipid-lowering therapy in subjects with type II diabetes: clinical effectiveness or clinical discretion? AB - Hypertension and lipid disorders in type II diabetes contribute to increased coronary risk, but optimal drug therapy has not been defined. We investigated primary care physicians choices of antihypertensive and lipid-lowering therapy for subjects with type II diabetes diagnosed with hypertension. Subjects were registered with 105 UK general practices in the General Practice Research Database and prescribed oral hypoglycaemic drugs for the first time between January 1993 and December 2001. We evaluated prescriptions for antihypertensive drugs in subjects with secondary diagnoses of hypertension in the first year following initiation of oral hypoglycaemic therapy. Data were analysed for 4519 diabetic subjects with diagnosed hypertension. Between 1993 and 2001, the proportion prescribed thiazide diuretics increased from 20 to 30%; angiotensin converting enzyme (ACE) inhibitors from 35 to 45% and angiotensin receptor blockers from 0 to 8%. The proportion of subjects prescribed lipid-lowering therapy increased from 8% in 1993 to 33% in 2001, with the proportion prescribed statins increasing from 1 to 30%. At different general practices, the proportion prescribed thiazide diuretics ranged from 0 to 52%, beta-blockers from 5 to 60%, ACE inhibitors from 15 to 81%, and statins from 0 to 50%. Variation between practices was not explained by adjusting for age, sex, prevalent coronary heart disease or study year. Trends in drug utilisation were consistent with the evolving evidence base but there were wide variations in drug utilisation between practices. A more consistent approach to drug selection might be associated with improved patient outcomes. PMID- 15361895 TI - Two dimensions of measurement error: classical and Berkson error in residential radon exposure assessment. AB - Measurement error in exposure assessment is unavoidable. Statistical methods to correct for such errors rely upon a valid error model, particularly regarding the classification of classical and Berkson error, the structure and the size of the error. We provide a detailed list of sources of error in residential radon exposure assessment, stressing the importance of (a) the differentiation between classical and Berkson error and (b) the clear definitions of predictors and operationally defined predictors using the example of two German case-control studies on lung cancer and residential radon exposure. We give intuitive measures of error size and present evidence on both the error size and the multiplicative structure of the error from three data sets with repeated measurements of radon concentration. We conclude that modern exposure assessment should not only aim to be as accurate and precise as possible, but should also provide a model of the remaining measurement errors with clear differentiation of classical and Berkson components. PMID- 15361896 TI - Disposable diaper to collect urine samples from young children for pyrethroid pesticide studies. AB - Disposable diapers are widely used in the US and many other areas in the world; therefore, they are ideal media for urine collection for measurement of young children's exposure to pesticides. However, disposable diapers normally contain polyacrylate polymers that make the extraction and analysis of urine very difficult. The objectives of this paper were to evaluate whether disposable diapers that contain polyacrylate granules can be extracted using salt solutions, and whether they can be used for the collection and quantitative measurements of selected urinary pyrethroid pesticide metabolites and creatinine. The storage stability of the metabolites and creatinine in a wet diaper at body temperature and at refrigeration temperature was also evaluated. Salt solutions including calcium chloride dihydrate, magnesium sulfate, ammonium acetate, and sodium chloride solutions were tested for efficiency of polymer shrinkage. Pyrethroid metabolites 3-(2,2-dichlorovinyl)-2,2-dimethyl-(1-cyclopropane) carboxylic acid (DCCA), 3-(2,2-dibromovinyl)-2,2,dimethyl-(1-cyclopropane) carboxylic acid (DBCA) and 3-phenoxybenzoic acid (3-PBA) were analyzed using LC/MS/MS and evaluated for recoveries in the urine released from the diapers. The study found calcium chloride dihydate to be satisfactory in releasing urine and metabolites from the polymers. The percent recoveries for the three tested pyrethroid metabolites were mostly in the range of 65-130. The percent recoveries for creatinine were in the range of 71-133. The detection limit for each of the three metabolites was 0.1 microg/l. The pyrethroid metabolites and creatinine were stable on the diaper for at least 72 h. We concluded from this study that calcium chloride dihydrate can successfully release urine and metabolites from polyacrylate-containing diapers, and the method is promising for studies of pyrethroid metabolites. PMID- 15361897 TI - Exposure assessment of male recruits in Slovenia to cadmium and lead due to biological monitoring. AB - AIMS: In Slovenia, there were no data available for an assessment of cadmium and lead levels in the population till 2001. The present study was conducted to evaluate the cadmium and lead burden from all sources (air, food, water) with the aim of obtaining initial information on cadmium and lead levels in blood of healthy and occupationally unexposed young males. METHODS: There were 464 participants included in the analysis of blood cadmium and lead levels, aged between 18 and 27 years, the median age being 20 years. Blood samples of maximum 2 ml were taken from the cubital vein for analysis of cadmium and lead levels. RESULTS: There were 463 blood samples analyzed for cadmium and 464 for lead. Of all tested persons, 38.2% had less than 0.5 microg/l of cadmium in their blood and 28.2% up to 1 microg/l. The median level of blood cadmium was 0.5 microg/l. The percentage of recruits with blood lead level over 100 microg was 3.1%. The median level of blood lead was 35 microg/l. The differences in blood levels of both pollutants were statistically significant with regard to the region of permanent residence (P<0.001). CONCLUSIONS: The results of the study give a good assessment of cadmium and lead burdening of young Slovene male population. The information obtained provides a foundation for further comparative research at the international level as well as for further study and risk assessment of individual population groups exposed to greater risk due to their lifestyle and eating habits. PMID- 15361898 TI - Comparing exposure metrics in the relationship between PM2.5 and birth weight in California. AB - Although studies suggest that air pollution is linked to perinatal outcomes, the geographic characterization of exposure to pollution differs between the studies. We compared neighborhood- and county-level measures of air pollution exposure, while examining the association between particulate matter less than 2.5 microm in aerodynamic diameter (PM(2.5)) and birth weight among full-term births in California in 2000. To reduce the effects of demographic variability, our analysis was limited to two populations of 8579 non-Hispanic white and 8114 Hispanic mothers who were married, between 20 and 30 years of age, completed at least a high school education, and gave birth for the first time. Measurements from the nearest monitor, and average and distance-weighted average of monitors within a 5-mile radius from each mother's residence (constituting neighborhood metrics) and the mean of monitors within each mother's county of residence were considered. PM(2.5) measurements, provided by the California Air Resources Board, were calculated to correspond to each mother's 9-month gestation period. Although metrics within the 5-mile radii and the county were highly correlated (r(2)=0.78), the county-level metric provided a stronger association between PM(2.5) and birth weight (beta=-4.04, 95% confidence interval =-6.71, -1.37) than the metric for the average of all monitors within 5-miles (beta=-1.38, 95% confidence interval =-3.36, 0.60) among non-Hispanic white mothers; similar results were observed among the Hispanic sample of mothers. Consequently, inferences from studies using different definitions of air pollution exposure may not be comparable. PMID- 15361899 TI - Evaluation of specific occupational asthma risks in a community-based study with special reference to single and multiple exposures. AB - OBJECTIVES: To compare two job exposure matrices (JEMs) for the evaluation of asthma risks related to specific occupational exposures in a community-based study. METHODS: A questionnaire on self-reported asthma, respiratory symptoms and current occupation was sent to the participants of the European Community Respiratory Health Survey in five areas in Spain. Both an asthma-specific JEM, including expert judgment steps and a general JEM, were applied to occupational codes. Risks of current asthma symptoms and wheeze in the last year associated with the obtained exposure estimates were evaluated. Correlations between specific exposures were investigated using explanatory factor analysis. RESULTS: Occupational exposures to the high-molecular-weight (MW) agents flour dust, enzymes, mites and animal-derived proteins as obtained by the asthma-specific JEM were positively associated with asthma outcomes. The effect of additional expert judgment steps was limited. High exposures to biological dust assessed by the general JEM without expert judgment was also associated with asthma. Many of the exposed individuals worked in environments with multiple exposures. CONCLUSIONS: Asthma risks associated with occupational exposures to specific high-MW agents could be identified from a population-based study using an asthma-specific JEM. Application of JEMs can be a useful tool to estimate asthma risks attributable to specific occupational exposures in the general population. However, these specific exposure risks should be interpreted in connection with the whole of concomitant exposures constituting the work environment. PMID- 15361900 TI - Comparison of spatial interpolation methods for the estimation of air quality data. AB - We recognized that many health outcomes are associated with air pollution, but in this project launched by the US EPA, the intent was to assess the role of exposure to ambient air pollutants as risk factors only for respiratory effects in children. The NHANES-III database is a valuable resource for assessing children's respiratory health and certain risk factors, but lacks monitoring data to estimate subjects' exposures to ambient air pollutants. Since the 1970s, EPA has regularly monitored levels of several ambient air pollutants across the country and these data may be used to estimate NHANES subject's exposure to ambient air pollutants. The first stage of the project eventually evolved into assessing different estimation methods before adopting the estimates to evaluate respiratory health. Specifically, this paper describes an effort using EPA's AIRS monitoring data to estimate ozone and PM10 levels at census block groups. We limited those block groups to counties visited by NHANES-III to make the project more manageable and apply four different interpolation methods to the monitoring data to derive air concentration levels. Then we examine method-specific differences in concentration levels and determine conditions under which different methods produce significantly different concentration values. We find that different interpolation methods do not produce dramatically different estimations in most parts of the US where monitor density was relatively low. However, in areas where monitor density was relatively high (i.e., California), we find substantial differences in exposure estimates across the interpolation methods. Our results offer some insights into terms of using the EPA monitoring data for the chosen spatial interpolation methods. PMID- 15361902 TI - Reduced-intensity non-T-cell depleted HLA-haploidentical stem cell transplantation for older patients based on the concept of feto-maternal tolerance. AB - With the recent progress in reduced-intensity conditioning stem cell transplantation (RIST) and taking into consideration the concept of feto-maternal immunological tolerance, we carried out non-T-cell depleted HLA haploidentical RIST from noninherited maternal antigen (NIMA) complementary siblings or offspring donors for four older patients: a patient with myeloplastic syndrome (MDS) and three patients with adult T-cell leukemia (ATL) in partial remission or with progressive disease. All patients showed early, durable engraftment, and no serious toxicities were observed apart from grade III mucositis in one case. Grade II acute GVHD occurred in two cases, which was well-controlled. In one ATL patient whose donor did not have NIMA microchimerism, tacrolimus could not be continued after engraftment due to renal dysfunction, and grade III acute GVHD (gut: stage 4) occurred on day 35. A patient with MDS was free from disease (requiring no transfusions and with a normal bone marrow) for 15 months. Two cases of ATL relapsed. Feto-maternal tolerance may lead to new RIST strategies in the haploidentical reduced-intensity situation, but further evaluation is required. PMID- 15361904 TI - Successful allogeneic haematopoietic stem cell transplantation in patients with poor-risk leukaemia and prior invasive fungal infection. PMID- 15361903 TI - Outcome of allogeneic bone marrow transplantation for children with advanced acute myeloid leukemia. AB - Allogeneic bone marrow transplantation (BMT) may offer the only chance of cure for children with acute myeloid leukemia (AML) in second complete remission (CR2) or with relapsed disease, but the outcome of these patients has not been clearly defined. We conducted a retrospective study of 58 children, median age 7.4 years (range 0.8-17.3), who received matched related or unrelated BMT at our institution for AML in CR2 (n = 12), in untreated first relapse (n = 11) or with refractory disease (n = 35), to identify risk factors associated with disease free survival (DFS). Life threatening to fatal regimen-related toxicity was observed in 22% of patients. Estimates of DFS at 5 years (95% confidence interval) for patients in CR2, with untreated first relapse and refractory disease were 58% (27-80%), 36% (11-63%) and 9% (2-21%), respectively. Non-relapse mortality estimates were 0%, 27% (0-54%) and 17% (5-30%), and relapse estimates were 42% (14-70%), 36% (8-65%) and 74% (60-89%), respectively. Advanced disease phase and cytogenetic abnormalities at the time of transplantation were each associated with decreased DFS and increased relapse in multivariable regression models. Survival for children transplanted in CR2 or untreated first relapse is higher than that previously reported, but relapse remains the major cause of treatment failure regardless of disease stage. PMID- 15361905 TI - West Nile virus infection in bone marrow transplant patients. PMID- 15361906 TI - Epstein-Barr virus-associated, CD20- polyclonal lymphoproliferative disorder after matched unrelated donor marrow transplantation. PMID- 15361907 TI - Cidofovir for treating adenoviral hemorrhagic cystitis in hematopoietic stem cell transplant recipients. AB - Adenovirus (AdV) infection is an important cause of morbidity and mortality in hematopoietic stem cell transplant (HSCT) recipients. We treated 16 patients with AdV hemorrhagic cystitis (HC) following HSCT with cidofovir (CDV; 1 mg/kg/day, three times weekly for 3 weeks). Patients included 10 males and six females with a median age of 50 years (range 10-62). Two of the 16 patients were unevaluable because of early death from nonadenoviral causes. CDV therapy cleared AdV from urine in 12 of 14 patients (86%). Of 14 patients, 10 (71%) showed clinical improvements in HC. Among 14 patients, seven (50%) had avoided renal damage, the most important CDV toxicity. One patient previously treated with foscarnet for cytomegalovirus (CMV) required hemodialysis, and CDV treatment was discontinued. In another patient, CDV treatment was discontinued because of grade 2 nephrotoxicity. Four patients became positive for CMV antigenemia while being treated with CDV, and two developed herpes simplex virus (HSV) stomatitis while being treated with CDV. CDV proved effective in treating AdV HC in transplant patients. However, CDV at 1 mg/kg/day given three times weekly failed to prevent breakthrough infection with CMV and HSV in some patients. PMID- 15361908 TI - Outcomes of unrelated cord blood transplants and allogeneic-related hematopoietic stem cell transplants in children with high-risk acute lymphocytic leukemia. AB - Acute lymphocytic leukemia (ALL) is a common indication for hematopoietic stem cell transplantation (HSCT) in children. Use of unrelated cord blood (UCB) has become increasingly popular as a stem cell source, given the rapid availability and decreased GVHD potential. Publications describing outcomes of children with leukemia who underwent UCB transplants have compared them to those having received unrelated donor marrow transplants. Results are similar. We compared our outcomes using UCB vs allogeneic-related hematopoietic stem cells in pediatric ALL patients since 1992. A total of 49 patients were analyzed. All patients were either in CR1 with high-risk features (n=21) or in CR2 (n=28) with initial remission less than 36 months. Patients received myeloablation with fractionated total body irradiation, cyclophosphamide, and etoposide and GVHD prophylaxis with cyclosporine and methotrexate. Antithymocyte globulin was added for UCB recipients to address the HLA differences. In all, 23 patients underwent allogeneic -related HSCT and 26 underwent UCB transplantation. Other than increased time to engraftment for UCB recipients, results are equivalent. The 3 year overall survival is 64% and 3-year event-free survival is 60% for both groups. Rates of GVHD and transplant-related mortality are also equivalent. UCB is a reasonable option for children with ALL who are referred for HSCT. PMID- 15361909 TI - Regimen-related toxicity following reduced-intensity stem-cell transplantation (RIST): comparison between Seattle criteria and National Cancer Center Common Toxicity Criteria (NCI-CTC) version 2.0. AB - Acute regimen-related toxicity (RRT) is minimal in reduced-intensity stem-cell transplantation (RIST). However, the Seattle RRT grading (Bearman et al), developed in the context of conventional-intensity transplantation, is frequently applied to RIST. We compared the National Cancer Institute Common Toxicity Criteria (NCI-CTC) version 2.0 with the Seattle criteria after RIST in 86 patients. RRT within 30 days of transplant graded by both sets of criteria were significantly associated with the outcome confirming the predictive value of both the systems. A total of 15 patients died of disease progression, and 12 of transplant-related mortality: RRT (n = 2), graft-versus-host disease (GVHD) (n = 7), infection (n = 1), and others (n = 2). GVHD-related deaths primarily resulted from infections after steroid treatment (n = 6) and bronchiolitis obliterans (n = 1). This study shows that NCI-CTC is appropriate in toxicity evaluation of RIST, and that its application to RIST enables a toxicity comparison between RIST and other types of cancer treatments. Since GVHD is a significant problem in RIST, modifications are required to evaluate immunological complications following RIST. PMID- 15361910 TI - The rationale behind autologous autoimmune hematopoietic stem cell transplant conditioning regimens: concerns over the use of total-body irradiation in systemic sclerosis. AB - Hematopoietic stem cell transplantation (HSCT) is becoming an increasingly recognized indication for treatment of autoimmune diseases and severe immune mediated disorders. However, multicenter registry data have demonstrated higher than anticipated early toxicity, approximately 10% for autoimmune diseases in general, and 20-27% for diffuse systemic sclerosis (scleroderma). If uncorrected, this high treatment-related mortality will hinder development of stem cell therapy for immune-mediated diseases. In order to develop safer regimens, we address some pitfalls and concepts involved in design and selection of conditioning regimens for autoimmune diseases in general, and because it is associated with the highest regimen-related toxicity, scleroderma in specific. PMID- 15361911 TI - Donor lymphocyte infusions for multiple myeloma: clinical results and novel perspectives. AB - Donor lymphocyte infusions (DLIs) provide effective therapy for patients with various hematological malignancies who have relapsed after allogeneic hematopoietic stem cell transplantation (HSCT). In patients with multiple myeloma (MM), DLIs can induce response rates of 40-52%. DLIs were employed as treatment for MM relapse or as prophylaxis for relapse in MM patients undergoing allo-HSCT. The clinically most relevant treatment-related morbidity with DLIs is the occurrence of graft-versus-host disease (GVHD). Secondly, graft failure and the immune escape of extramedullary plasmocytoma have been reported. The fact that previous clinical reports have documented graft-versus-myeloma (GVM) activity without GVHD suggests that at least two distinct immunocompetent cell populations mediating GVHD and/or GVM may exist. Further characterization of the effector cells such as T cells and/or NK cells and their targets may help to clarify the immune response that mediates the GVM effect. This review considers the results of clinical approaches with DLI for MM, with emphasis on strategies to prevent GVHD while preserving the GVM effect. Furthermore, currently investigated molecular antigenic targets for the GVM effect such as MM-specific idiotypic determinant of immunoglobulin variable regions, several PRAME epitopes and antigenic structures encoded by cancer germline-specific genes as candidates for immunotherapy trials are discussed. PMID- 15361912 TI - Evidence of graft-versus-tumour effect following allogeneic haematopoietic stem cell transplantation in renal cancer other than clear cell type. PMID- 15361913 TI - An update on graft-versus-host and graft-versus-leukemia reactions: a summary of the sixth International Symposium held in Schloss Ellmau, Germany, January 22-24, 2004. AB - The Sixth International Symposium on Graft-versus-Host and Graft-versus Leukemia Reactions was held in Schloss Ellmau (near Garmisch-Partenkirchen, Germany) between January 21 and 24, 2004. A total of 110 invited participants (scientists and clinicians working in the area of allogeneic stem cell transplantation) discussed current topics. Major topics of the 2004 meeting were: clinical results of donor lymphocyte infusions, basic biology, immunogenetics, function and clinical relevance of natural killer cells, haplo-identical stem cell transplantation, immune monitoring and immune modulation. Further highlights were: adoptive immunotherapy, vaccination and antibody-mediated strategies. As can be seen in the summaries of the individual presentations, important advances have occurred in our understanding of GVH and GVL reactions. Each session was followed by an animated discussion, which resulted in new ideas, insights and projects both for basic research and clinical transplantation. This year's symposium ('From Marrow Transplantation to Cell Therapy') was jointly organized by the Ludwigs-Maximilians-University of Munich (Sonderforschungsbereich 455), GSF (National Research Center for Environment and Health) and the EBMT Immunobiology Working Party. The organizers and authors of the conference proceedings would like to extend their gratitude to all participants for sharing their ideas, slides and manuscripts and making this event possible. PMID- 15361914 TI - Pericarditis occurring with engraftment syndrome in a thalassemic patient. PMID- 15361915 TI - Fusarium dimerum infection in a stem cell transplant recipient treated successfully with voriconazole. AB - We report the first case, to our knowledge, of a proven Fusarium dimerum soft tissue infection in a stem cell transplant recipient treated successfully with voriconazole. There is a well-documented increase in the incidence, diversity and antifungal resistance of invasive mould infections in the immunocompromised patient population. The management of these infections is changing as new, more efficacious and less toxic antifungal agents become available. We present the case of a 19-year-old female diagnosed with a proven F. dimerum soft-tissue infection of the foot and possible pulmonary infection with the same organism 10 days following a sibling allogeneic stem cell transplant for severe aplastic anaemia. The infection developed despite treatment with 3 mg/kg AmBisome for a concurrent chest infection. She was treated successfully with voriconazole. PMID- 15361916 TI - Clinical impact of graft-versus-host disease against leukemias not in remission at the time of allogeneic hematopoietic stem cell transplantation from related donors. The Japan Society for Hematopoietic Cell Transplantation Working Party. AB - Acute graft-versus-host disease (GVHD) increases post-transplant mortality and morbidity, but exerts a potent graft-versus-leukemia (GVL) effect. To clarify the impact of GVHD on outcome after transplant in aggressive diseases, patients with acute myeloid or lymphoblastic leukemia (AML, n = 366 or ALL, n = 255) in nonremission states, or chronic myelogenous leukemia (CML, n = 180) in accelerated phase (AP) or blastic crisis (BC), who received allogeneic hematopoietic stem cell transplantation (HSCT) from a related donor between 1991 and 2000, were analyzed. Significant improvement in overall and disease-free survival (DFS) was detected with grade I acute GVHD in AML (P = 0.0002 for overall survival and 0.0009 for DFS, respectively) and in CML (P = 0.0256 and 0.0366, respectively), while the trend towards improved survival was observed in ALL. Relapse rate was lower in grade I acute GVHD than in grade II in all three diseases, suggesting that treatment for grade II GVHD may compromise the GVL effect associated with GVHD. Chronic GVHD was found to suppress relapse in CML and ALL, but not in AML, although no improvement in survival was observed in any disease category. Our results suggest that treatment for grade II acute GVHD may need to be attenuated in transplant for refractory leukemias. PMID- 15361917 TI - Knowledge About Recommended Treatment and Management of Major Depressive Disorder, Panic Disorder, and Generalized Anxiety Disorder Among Family Physicians. AB - BACKGROUND: Concerns have been raised about whether primary care physicians appropriately manage mental disorders. We assessed family physicians' knowledge of appropriate management of major depressive disorder (MDD), panic disorder, and generalized anxiety disorder (GAD). METHOD: Active members of the Texas Academy of Family Physicians (N = 3553) were mailed a questionnaire in 2002 asking them to indicate which treatments they felt were effective for MDD, panic disorder, and GAD and also to indicate how they had treated their last patient with each disorder. Their treatment strategies were then compared with current guidelines. RESULTS: 574 physicians (16%) responded. The percentage of respondents scoring at or above 80% for knowledge of effective treatments was 88.3% for MDD, 16.8% for panic disorder, and 12.5% for GAD (p <.001 for MDD vs. panic disorder or GAD). Only 0.3% of MDD patients, 1.4% of panic disorder patients, and 4.0% of GAD patients were not prescribed at least 1 of the effective treatments. Referral rates to mental health providers were high for all 3 conditions. CONCLUSIONS: There were significant gaps in physician knowledge of current guidelines on treating panic disorder and GAD, but not MDD. However, most patients with one of the disorders were either referred to a mental health provider or treated with an effective modality. PMID- 15361918 TI - The Metabolic Syndrome in Patients With Severe Mental Illnesses. AB - BACKGROUND: Since the introduction of the first atypical antipsychotics in the early 1990s, this class of medication has been increasingly relied upon for the treatment of a variety of patients with psychotic and mood disorders.DATA SOURCES: The following retrospective review was derived from the MEDLINE database using the search terms metabolic syndrome, insulin resistance, obesity, diabetes, severe mental illness, schizophrenia, bipolar disorder, mood disorders, depression, unipolar depression, and prevalence from 1966 to the present. LITERATURE SYNTHESIS: Coincident with the growing usage of these agents, there have been a growing number of literature reports of changes in metabolic homeostasis among patients taking these medications. These changes have led to interest in evaluating whether there is a relationship among these mental illnesses, their psychiatric treatments, and certain physical comorbidities known collectively as the metabolic syndrome. This article reviews the existing literature around the metabolic syndrome in patients with severe mental illnesses. CONCLUSION: Patients with severe mental illnesses, particularly schizophrenia and chronic mood disorders, demonstrate a higher prevalence of metabolic syndrome or its components compared with the general population. Based upon this increased risk in these patients, baseline and periodic medical evaluations should become a standard component in ongoing clinical assessment. PMID- 15361920 TI - Recognizing and Treating the Physical Symptoms of Depression in Primary Care. AB - This Academic Highlights section of The Primary Care Companion to The Journal of Clinical Psychiatry presents the highlights from the teleconference "Recognizing the Physical Symptoms of Depression," which was held March 16, 2004. The teleconference and this ACADEMIC HIGHLIGHTS were independently developed pursuant to an unrestricted educational grant from Eli Lilly and Company. This report was prepared by Physicians Postgraduate Press, Inc. PMID- 15361919 TI - A Review of the Neuropharmacology of Bupropion, a Dual Norepinephrine and Dopamine Reuptake Inhibitor. AB - BACKGROUND: The neurochemical and biological effects of antidepressant medications have become better defined over the last decade. When the anti depressant bupropion was introduced in the United States in 1989, the specific pharmacologic basis of its clinical effects was uncertain. Research conducted over the past decade has significantly advanced the understanding of the neuropharmacology of bupropion and has demonstrated a novel mechanism of antidepressant activity. This article discusses the mechanism of action of bupropion and relates the drug's neuropharmacologic effects to its clinical efficacy and tolerability profiles. DATA SOURCES: Data were obtained via the MEDLINE database in an English-language search spanning the period 1965 to May 2002 and using the search terms bupropion, bupropion SR, and antidepressants, as well as from the manufacturer's bupropion databases. CONCLUSIONS: The preclinical and clinical data show that bupropion acts via dual inhibition of norepinephrine and dopamine reuptake and is devoid of clinically significant serotonergic effects or direct effects on postsynaptic receptors. Dual norepinephrine and dopamine reuptake inhibition is associated with a unique clinical profile. Bupropion has demonstrated efficacy comparable to that of other antidepressants. However, because bupropion is a selective norepinephrine and dopamine reuptake inhibitor with no serotonergic activity, common antidepressant-associated side effects, such as sexual dysfunction, weight gain, and sedation, are not associated with bupropion therapy. PMID- 15361921 TI - Making a Difference. PMID- 15361922 TI - A Doctor's Life Can Sometimes Be Hairy. PMID- 15361923 TI - A Case Report of Fluoxetine- and Venlafaxine-Induced Hair Loss. PMID- 15361924 TI - The Benefits of Exercise for the Clinically Depressed. AB - Millions of Americans suffer from clinical depression each year. Most depressed patients first seek treatment from their primary care providers. Generally, depressed patients treated in primary care settings receive pharmacologic therapy alone. There is evidence to suggest that the addition of cognitive-behavioral therapies, specifically exercise, can improve treatment outcomes for many patients. Exercise is a behavioral intervention that has shown great promise in alleviating symptoms of depression. The current review discusses the growing body of research examining the exercise-depression relationship that supports the efficacy of exercise as an adjunct treatment. Databases searched were Medline, PsycLit, PubMed, and SportsDiscus from the years 1996 through 2003. Terms used in the search were clinical depression, depression, exercise, and physical activity. Further, because primary care physicians deliver important mental health services to the majority of depressed patients, several specific recommendations are made regarding counseling these patients on the adoption and maintenance of exercise programs. PMID- 15361925 TI - The Association Between Spirituality and Depression in an Urban Clinic. AB - OBJECTIVE: To investigate the correlation between spiritual beliefs and depression in an urban population. METHOD: A convenience sample of adult patients of an urban primary care clinic completed a self-administered questionnaire consisting of the Zung Depression Scale and the Spiritual Involvement and Beliefs Scale (SIBS). RESULTS: Among 122 respondents, 99 (81%) reported that they consider themselves religious. Responses from the Zung Depression Scale found that 76 (62%) of the patients were depressed and 46 (38%) were not. The Pearson correlation coefficient between the Zung Depression Scale and the SIBS was -0.36 (p <.0001). Backward stepwise regression analysis revealed that SIBS score and physical health predicted the Zung Depression Scale score. Age, gender, ethnicity, religious affiliation, and income showed no significant association with depression. Analysis of individual SIBS items revealed that high spirituality scores on items in the domain of intrinsic beliefs, such as belief in a higher power (p <.01), the importance of prayer (p <.0001), and finding meaning in times of hardship (p <.05), were associated negatively with depression. Attendance of religious services had no significant association with depression. CONCLUSION: Appropriate encouragement of a patient's spiritual beliefs may be a helpful adjunct to treating depression. PMID- 15361926 TI - Murug, Waali, and Gini: Expressions of Distress in Refugees From Somalia. AB - OBJECTIVE: To study how mental illness is understood, expressed, and treated among Somali refugees and how these factors influence use of health services for mental problems. METHOD: Seventeen adult Somali refugees (9 women, 8 men) were recruited by mail or by word-of-mouth to participate in the study. The study setting was an urban community health center in Rochester, N.Y., that provides family practice patient care to local Somali refugees. A qualitative design was used that incorporated a combination of methods, chiefly semistructured interviews. Interviews focused on the ways in which sadness, depression, and anxiety are expressed and on the participants' understanding of the origins of and treatment strategies for these problems. Interview transcripts were analyzed to identify recurrent themes. RESULTS: Nearly all participants felt that mental illness was a new problem for their community that did not exist to the same extent in prewar Somalia. Themes that emerged to explain the causes of mental illness included the shock and devastation of war; dead, missing, or separated family members; and spirit possession or a curse. Three major types of mental problems were identified that were associated with specific behaviors and treatment strategies: murug (sadness or suffering), gini (craziness due to spirit possession), and waali (craziness due to severe trauma). Rather than seek help from a clinician, participants preferred to first use family support, prayer, or traditional therapies for most situations. CONCLUSION: Somali refugees have distinct ways of conceptualizing, expressing, and treating commonly understood mental problems. Participants differed in their opinions about whether they would consult a doctor to discuss feelings of sadness or craziness. Effective mental health care of refugees should address culture-specific belief systems in diagnosis and treatment. PMID- 15361927 TI - Role of Cholinesterase Inhibitors in Managing Behavioral Problems in Alzheimer's Disease. AB - Alzheimer's disease is characterized by progressive cognitive and functional decline and the emergence of behavioral disturbances. Behavioral symptoms, in particular, cause great distress to caregivers, creating an emotional and financial burden that often prompts the caregiver to place the patient in a nursing facility. The clinical deterioration in Alzheimer's disease is, in part, a result of deficits involving several neurochemical pathways. The cholinergic system, which is the most consistently and dramatically affected neurotransmitter system in Alzheimer's disease, has been strongly implicated in the emergence of neuropsychiatric symptoms. This article reviews evidence suggesting that, in addition to effects on cognition and function, the cholinesterase inhibitors benefit the behavioral symptoms of Alzheimer's disease. Pharmacologic and nonpharmacologic treatment strategies for the management of behavioral symptoms are discussed. PMID- 15361928 TI - Cognitive Therapy for Dysthymia. PMID- 15361929 TI - What Hands! PMID- 15361930 TI - Banned, but Not Forgotten: A Case of Ephedrine-Induced Psychosis. PMID- 15361931 TI - Analysis of among-site variation in substitution patterns. AB - Substitution patterns among nucleotides are often assumed to be constant in phylogenetic analyses. Although variation in the average rate of substitution among sites is commonly accounted for, variation in the relative rates of specific types of substitution is not. Here, we review details of methodologies used for detecting and analyzing differences in substitution processes among predefined groups of sites. We describe how such analyses can be performed using existing phylogenetic tools, and discuss how new phylogenetic analysis tools we have recently developed can be used to provide more detailed and sensitive analyses, including study of the evolution of mutation and substitution processes. As an example we consider the mitochondrial genome, for which two types of transition deaminations (C-->T and A-->G) are strongly affected by single-strandedness during replication, resulting in a strand asymmetric mutation process. Since time spent single-stranded varies along the mitochondrial genome, their differential mutational response results in very different substitution patterns in different regions of the genome. PMID- 15361933 TI - Genetic response to climatic change: insights from ancient DNA and phylochronology. AB - Understanding how climatic change impacts biological diversity is critical to conservation. Yet despite demonstrated effects of climatic perturbation on geographic ranges and population persistence, surprisingly little is known of the genetic response of species. Even less is known over ecologically long time scales pertinent to understanding the interplay between microevolution and environmental change. Here, we present a study of population variation by directly tracking genetic change and population size in two geographically widespread mammal species (Microtus montanus and Thomomys talpoides) during late Holocene climatic change. We use ancient DNA to compare two independent estimates of population size (ecological and genetic) and corroborate our results with gene diversity and serial coalescent simulations. Our data and analyses indicate that, with population size decreasing at times of climatic change, some species will exhibit declining gene diversity as expected from simple population genetic models, whereas others will not. While our results could be consistent with selection, independent lines of evidence implicate differences in gene flow, which depends on the life history strategy of species. PMID- 15361932 TI - Preserving genome integrity: the DdrA protein of Deinococcus radiodurans R1. AB - The bacterium Deinococcus radiodurans can withstand extraordinary levels of ionizing radiation, reflecting an equally extraordinary capacity for DNA repair. The hypothetical gene product DR0423 has been implicated in the recovery of this organism from DNA damage, indicating that this protein is a novel component of the D. radiodurans DNA repair system. DR0423 is a homologue of the eukaryotic Rad52 protein. Following exposure to ionizing radiation, DR0423 expression is induced relative to an untreated control, and strains carrying a deletion of the DR0423 gene exhibit increased sensitivity to ionizing radiation. When recovering from ionizing-radiation-induced DNA damage in the absence of nutrients, wild-type D. radiodurans reassembles its genome while the mutant lacking DR0423 function does not. In vitro, the purified DR0423 protein binds to single-stranded DNA with an apparent affinity for 3' ends, and protects those ends from nuclease degradation. We propose that DR0423 is part of a DNA end-protection system that helps to preserve genome integrity following exposure to ionizing radiation. We designate the DR0423 protein as DNA damage response A protein. PMID- 15361934 TI - Genetic analysis of pathways regulated by the von Hippel-Lindau tumor suppressor in Caenorhabditis elegans. AB - The von Hippel-Lindau (VHL) tumor suppressor functions as a ubiquitin ligase that mediates proteolytic inactivation of hydroxylated alpha subunits of hypoxia inducible factor (HIF). Although studies of VHL-defective renal carcinoma cells suggest the existence of other VHL tumor suppressor pathways, dysregulation of the HIF transcriptional cascade has extensive effects that make it difficult to distinguish whether, and to what extent, observed abnormalities in these cells represent effects on pathways that are distinct from HIF. Here, we report on a genetic analysis of HIF-dependent and -independent effects of VHL inactivation by studying gene expression patterns in Caenorhabditis elegans. We show tight conservation of the HIF-1/VHL-1/EGL-9 hydroxylase pathway. However, persisting differential gene expression in hif-1 versus hif-1; vhl-1 double mutant worms clearly distinguished HIF-1-independent effects of VHL-1 inactivation. Genomic clustering, predicted functional similarities, and a common pattern of dysregulation in both vhl-1 worms and a set of mutants (dpy-18, let-268, gon-1, mig-17, and unc-6), with different defects in extracellular matrix formation, suggest that dysregulation of these genes reflects a discrete HIF-1-independent function of VHL-1 that is connected with extracellular matrix function. PMID- 15361935 TI - Population history and natural selection shape patterns of genetic variation in 132 genes. AB - Identifying regions of the human genome that have been targets of natural selection will provide important insights into human evolutionary history and may facilitate the identification of complex disease genes. Although the signature that natural selection imparts on DNA sequence variation is difficult to disentangle from the effects of neutral processes such as population demographic history, selective and demographic forces can be distinguished by analyzing multiple loci dispersed throughout the genome. We studied the molecular evolution of 132 genes by comprehensively resequencing them in 24 African-Americans and 23 European-Americans. We developed a rigorous computational approach for taking into account multiple hypothesis tests and demographic history and found that while many apparent selective events can instead be explained by demography, there is also strong evidence for positive or balancing selection at eight genes in the European-American population, but none in the African-American population. Our results suggest that the migration of modern humans out of Africa into new environments was accompanied by genetic adaptations to emergent selective forces. In addition, a region containing four contiguous genes on Chromosome 7 showed striking evidence of a recent selective sweep in European-Americans. More generally, our results have important implications for mapping genes underlying complex human diseases. PMID- 15361937 TI - Continued colonization of the human genome by mitochondrial DNA. AB - Integration of mitochondrial DNA fragments into nuclear chromosomes (giving rise to nuclear DNA sequences of mitochondrial origin, or NUMTs) is an ongoing process that shapes nuclear genomes. In yeast this process depends on double-strand-break repair. Since NUMTs lack amplification and specific integration mechanisms, they represent the prototype of exogenous insertions in the nucleus. From sequence analysis of the genome of Homo sapiens, followed by sampling humans from different ethnic backgrounds, and chimpanzees, we have identified 27 NUMTs that are specific to humans and must have colonized human chromosomes in the last 4-6 million years. Thus, we measured the fixation rate of NUMTs in the human genome. Six such NUMTs show insertion polymorphism and provide a useful set of DNA markers for human population genetics. We also found that during recent human evolution, Chromosomes 18 and Y have been more susceptible to colonization by NUMTs. Surprisingly, 23 out of 27 human-specific NUMTs are inserted in known or predicted genes, mainly in introns. Some individuals carry a NUMT insertion in a tumor-suppressor gene and in a putative angiogenesis inhibitor. Therefore in humans, but not in yeast, NUMT integrations preferentially target coding or regulatory sequences. This is indeed the case for novel insertions associated with human diseases and those driven by environmental insults. We thus propose a mutagenic phenomenon that may be responsible for a variety of genetic diseases in humans and suggest that genetic or environmental factors that increase the frequency of chromosome breaks provide the impetus for the continued colonization of the human genome by mitochondrial DNA. PMID- 15361936 TI - A Drosophila pattern recognition receptor contains a peptidoglycan docking groove and unusual L,D-carboxypeptidase activity. AB - The Drosophila peptidoglycan recognition protein SA (PGRP-SA) is critically involved in sensing bacterial infection and activating the Toll signaling pathway, which induces the expression of specific antimicrobial peptide genes. We have determined the crystal structure of PGRP-SA to 2.2-A resolution and analyzed its peptidoglycan (PG) recognition and signaling activities. We found an extended surface groove in the structure of PGRP-SA, lined with residues that are highly diverse among different PGRPs. Mutational analysis identified it as a PG docking groove required for Toll signaling and showed that residue Ser158 is essential for both PG binding and Toll activation. Contrary to the general belief that PGRP SA has lost enzyme function and serves primarily for PG sensing, we found that it possesses an intrinsic L,D-carboxypeptidase activity for diaminopimelic acid-type tetrapeptide PG fragments but not lysine-type PG fragments, and that Ser158 and His42 may participate in the hydrolytic activity. As L,D-configured peptide bonds exist only in prokaryotes, this work reveals a rare enzymatic activity in a eukaryotic protein known for sensing bacteria and provides a possible explanation of how PGRP-SA mediates Toll activation specifically in response to lysine-type PG. PMID- 15361938 TI - Multicenter surveillance of antimicrobial resistance of Streptococcus pyogenes, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis to 14 oral antibiotics. AB - BACKGROUND AND PURPOSE: Data on the in vitro activities of orally administered cephalosporins, particularly third-generation cephalosporins, against recent pathogens responsible for community-respiratory tract infection are lacking. METHODS: A susceptibility surveillance of 267 isolates of Streptococcus pneumoniae, 205 of Streptococcus pyogenes, 204 of Haemophilus influenzae, and 147 of Moraxella catarrhalis to 14 oral antimicrobial agents using the agar dilution method was carried out from March 2002 to October 2002 in Taiwan. RESULTS: High rates of non-susceptibility to penicillin (60%), cefaclor (67%), cefuroxime (62%), cefpodoxime (64%), clarithromycin (91%), and trimethoprim-sulfamethoxazole (98%) for S. pneumoniae isolates and high rates of non-susceptibility to ampicillin (70%), clarithromycin (34%), and trimethoprim-sulfamethoxazole (63%) for H. influenzae isolates were found. The rank order of oral cephalosporin activity based on the minimum concentrations at which 90% of the isolates were inhibited (MIC90s) for S. pneumoniae was cefpodoxime > cefuroxime > cefixime > cefaclor, cephradine > cephalexin and for H. influenzae and M. catarrhalis was cefixime > cefpodoxime > cefuroxime > cefaclor > cephalexin, cephradine. Among the 75 S. pneumoniae isolates resistant to penicillin (MICs ranged 2 to 4 mg/L), 4% were intermediate to amoxicillin and > 90% were resistant to cefaclor, cefuroxime, and cefpodoxime. For S. pyogenes isolates, all were susceptible to penicillin, 21% were not susceptible to clarithromycin and 4% were not susceptible to clindamycin. Thirty four percent of H. influenzae isolates were not susceptible to clarithromycin. The MIC90 of clarithromycin against M. catarrhalis isolates was 0.5 mg/L. CONCLUSIONS: Cefpodoxime, cefixime, and cefuroxime are promising agents against these bacterial pathogens, except for penicillin-non-susceptible S. pneumoniae isolates. PMID- 15361939 TI - Drug resistance pattern of Mycobacterium tuberculosis in a university hospital in Taiwan, 1998-2002. AB - BACKGROUND AND PURPOSE: During 1998-2002, most specialized tuberculosis (TB) hospitals in Taiwan were closed; as a result, more TB patients are being managed in general hospital settings. This study investigated the prevalence, patterns and risk factors of drug-resistant Mycobacterium tuberculosis at a university hospital in the 5 years after decentralization of the TB administrative and clinical control infrastructure which occurred during the implementation of the national health insurance system in Taiwan. METHODS: A total of 1411 initial isolates of M. tuberculosis from specimens collected during January 1998 through December 2002 were tested for drug susceptibility of first-line anti-TB drugs using the agar proportional method. RESULTS: The numbers of newly diagnosed culture-positive TB patients increased from 139 in 1998 to 380 in 2002. The drug resistance pattern of M. tuberculosis among these isolates was as follows: 268 (19.0%) strains were resistant to isoniazid, 86 (6.1%) to rifampin, 221 (15.7%) to ethambutol, 141 (10.0%) to streptomycin, and 430 (30.5%) to 1 of these 4 drugs. Multidrug resistance (MDR), i.e., resistance to at least rifampin and isoniazid, was observed in 72 isolates (5.1%). Of the 1411 patients, isolates from patients with age < 65 years had a higher multiple drug resistance rate than those from patients with age > or = 65 years (57/781, 7.3% vs 15/630, 2.4%; p < 0.001). In the analysis of risk factors for MDR, patients with MDR isolates had a significantly higher incidence of previous TB history, anti-TB therapy, longer duration of symptoms, cavitary lesions in chest X-ray, and mortality. CONCLUSION: A dramatic increase in cases of TB among patients treated at this university hospital was seen after the decentralization of the TB control infrastructure in Taiwan. The prevalence of drug resistance in isolates from culture-positive TB patients was 30.5% and the prevalence of MDR was 5.1%. PMID- 15361940 TI - Efficacy of gabexate mesilate on disseminated intravascular coagulation as a complication of infection developing after abdominal surgery. AB - BACKGROUND AND PURPOSE: Gabexate mesilate (GM) is a promising anticoagulation treatment for disseminated intravascular coagulation (DIC). This study was designed to examine the effect of GM on DIC associated with the development of infection after abdominal surgery in the intensive care unit (ICU). METHODS: From January 1999 to March 2002, 50 consecutive ICU patients suffering DIC associated with the development of infection after abdominal surgery were enrolled in this study. Twenty five of the patients were randomized to receive treatment with GM by central intravenous infusion at 1 mg/kg/hour for 5 days or longer, while the remaining 25 were not treated. Blood clotting tests were performed and cytokine levels including tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were examined on days 1, 3, and 7 after admission. General blood tests and D dimer tests were conducted before and after GM administration. The gender, age, mortality, Acute Physiology and Chronic Health Evaluation II (APACHE-II) scores and severity of DIC were compared between the 2 groups. RESULTS: No significant difference was found between the 2 groups in TNF-alpha and IL-6 concentration on days 1, 3, and 7. The mortality rate was similar between the 2 groups. However, DIC and APACHE-II scores were significantly lower in the GM-treated patients than in controls. CONCLUSION: In this study, GM (1 mg/kg/hour) did not reduce the concentration of TNF-alpha and IL-6, or alter the mortality rate in patients with DIC resulting from infectious complications after surgery. Nevertheless, APACHE II scores indicated that GM reduced the DIC severity and improved the clinical condition of patients. PMID- 15361941 TI - Characteristics and trends in incidence of inflammatory bowel disease in Taiwanese children. AB - BACKGROUND AND PURPOSE: Although chronic inflammatory bowel disease (IBD) is rare in Asian children, increasing numbers of pediatric patients with chronic IBD worldwide have been noted in recent years. This study was conducted to delineate the trend in incidence and clinical patterns of childhood IBD in Taiwan. METHODS: All children admitted to National Taiwan University Hospital (NTUH) between 1979 and 2000 who met the criteria for IBD, Crohn's disease (CD), ulcerative colitis (UC), probable CD (PCD), or indeterminate colitis were included. The clinical features and outcomes were analyzed retrospectively. Incidence was calculated using cases of chronic diarrhea during the same period of time as the risk population. RESULTS: IBD was diagnosed in 17 children (9 females and 8 males, aged 2 months to 18 years) during the study period. Six (35%) of these children had UC, 9 (53%) had CD, and 2 (12%) had PCD. The cumulative incidence of CD during 1979-1995 was 0.85%, and increased to 2.6% during 1996-2000 (p < 0.001), while the incidence of UC did not change significantly between these periods (from 0.85% to 0.99%, p = 0.16). The median interval from onset to diagnosis was 7.7 months. Eighty percent of patients had moderate to severe disease activity at diagnosis. The follow-up duration ranged from less than 1 year to 20 years, with a mean of 4.3 years. Two patients were lost to follow-up. Eighty six percent of patients responded to treatment, and 80% of patients had inactive to mild disease activity when re-evaluated at the end of 2000. CONCLUSION: There has been a marked recent increase in the incidence of childhood CD in Taiwan but the rate of childhood UC has remained unchanged. Eighty percent of cases of childhood IBD responded well to treatment. PMID- 15361942 TI - Intravitreal triamcinolone injection for macular edema secondary to increased retinal vascular permeability. AB - BACKGROUND AND PURPOSE: Macular edema is an important cause of visual loss in various retinal diseases, and may be refractory to conventional treatment. We investigated the efficacy of intravitreal injection of triamcinolone acetonide for the treatment of intractable macular edema. METHODS: A prospective study was conducted in 17 patients (18 eyes) with a diagnosis of macular edema unresponsive to conventional treatment, resulting from inflammatory or retinal vascular diseases. The underlying diseases associated with the development of macular edema were: diabetic retinopathy (6 eyes), branch retinal vein occlusion (5 eyes), Irvine-Gass syndrome (3 eyes), and central retinal vein occlusion (4 eyes). Triamcinolone acetonide 4 mg was injected intravitreally. Ophthalmological examinations, fundus photography, fluorescein angiography, and optical coherence tomography were performed before treatment, 3 months and 6 months after treatment. During the follow-up period, recurrent macular edema was retreated with the same dosage and followed for another 6 months. RESULTS: Sixteen eyes received a single injection and 2 others (11%) underwent reinjection after the 3 month follow-up examination. Among the eyes that received a single injection, 10 (63%) showed visual acuity gain of 2 or more Snellen lines at the 6-month follow up. The pretreatment central macular thickness averaged 581 microm [corrected] and reduced to 215 microm [corrected] at the 6-month follow-up. In the 2 eyes that received reinjection, macular edema also showed significant reduction 6 months after retreatment. Sterile endophthalmitis was noted in 3 eyes (17%). Two eyes (11%) developed significant cataract. Three eyes (17%) developed high intraocular pressure; 1 of them (5.5%) ultimately required filtering surgery. CONCLUSIONS: Intravitreal injection of triamcinolone temporarily reduced refractory macular edema. Potential complications should be monitored after treatment. PMID- 15361943 TI - Long-term outcome of bulimia nervosa in Taiwanese. AB - BACKGROUND AND PURPOSE: Patients with bulimia nervosa (BN) often experience many associated psychiatric symptoms and behaviors other than disordered eating and body image disturbance. Follow-up studies showed a broad range of remission rates and few prognostic factors were consistently identified. This study examined the outcome of BN in Taiwanese patients using a broad scope of assessment methods. METHODS: From January 1985 to August 1999, a total of 68 consecutive patients with BN who visited the Department of Psychiatry at National Taiwan University Hospital were identified by chart review and invited to participate. Forty five patients (66.2%) agreed to participate in the study and were invited back for a follow-up assessment. The follow-up assessments used both semi-structured interview and self-rating questionnaires. RESULTS: The mean duration of follow-up was 4.7 +/- 2.9 (2 to 15) years. One patient died of gradual weight loss and malnutrition during the follow-up period. Forty four percent of the patients (n = 20) had no signs of eating disorder at follow-up, while 56% of the patients (n = 25) still met the criteria for diagnosis of an eating disorder (anorexia nervosa, BN, and eating disorders not otherwise specified). The rate of crossover from BN to anorexia nervosa was 6.6% (n = 3). The illness persisted for more than 2 years in most of the patients (84.5%). Purging subtype of BN was the only predictor of the presence of an eating disorder diagnosis at follow-up. CONCLUSIONS: This study found a high percentage of Taiwanese patients with BN failed to recover from eating disorders on long-term follow-up. The need for attention to the chronic nature of BN should be emphasized by medical professionals managing these patients. PMID- 15361944 TI - Primary Sjogren's syndrome complicated with cryoglobulinemic glomerulonephritis, myocarditis, and multi-organ involvement. AB - Glomerulonephritis in primary Sjogren's syndrome is rarely reported. Cryoglobulinemic glomerulonephritis with the presence of cryoglobulin deposition in the glomerular capillary lumen in primary Sjogren's syndrome is extremely rare. A 51-year-old woman with primary Sjogren's syndrome for > 10 years complained of fever, hypertension, and proteinuria. In addition, novel manifestations, including myocarditis with heart failure, pericardial effusion, and polyneuropathy (sensory motor neuropathy) were also noted. Cryoglobulinemia test was positive, and kidney biopsy results were consistent with cryoglobulinemic glomerulonephritis. There were no symptoms associated with systemic lupus erythematosus or other connective tissue disease. Treatment with monthly methylprednisolone and cyclophosphamide pulse therapy for 6 months resulted in resolution of proteinuria, heart failure, and neurologic symptoms. PMID- 15361945 TI - Monostotic fibrous dysplasia of the thoracic spine with malignant transformation. AB - Monostotic fibrous dysplasia involving the spine is rare. There have been only 6 previous reports of monostotic fibrous dysplasia involving the thoracic spine and none had malignant transformation. We report a case of monostotic fibrous dysplasia of the thoracic spine with malignant transformation. Findings on plain radiographs, isotope bone scan, computed tomography and magnetic resonance imaging are discussed and compared with those of the previously reported cases. PMID- 15361946 TI - A mucolipidosis III patient presenting characteristic sonographic and magnetic resonance imaging findings of claw hand deformity. AB - Mucolipidosis III (ML-III), or pseudo-Hurler polydystrophy, is an autosomal recessive Hurler-like disorder without mucopolysacchariduria. The diagnosis is challenging for rheumatologists since the musculoskeletal presentation is similar to some rheumatic diseases. We report a case of ML-III in a 16-year-old Taiwanese boy. The characteristic findings of sonography and magnetic resonance imaging (MRI) of claw hand deformity are described. A 16-year-old boy was referred to our rheumatologic clinic because of progressive claw hand deformity, multiple joint stiffness and tightness of the skin over the fingers at the age of 6 years. Sonography and MRI examination disclosed tendon sheath thickening over extensor tendons of both wrists and fingers without features of active inflammation over tendons or joints nor thickening of skin. Urinary glycosaminoglycans were normal. The diagnosis of ML-III was confirmed by the presence of elevated activities of beta-glucuronidase (2141.99 nmol/mg protein/hour), arylsulfatase A (1237.7 nmol/mg protein/hour) and alpha-fucosidase (52.95 nmol/mg protein/hour) in his plasma and decreased activity of these lysosomal enzymes in cultured skin fibroblasts. Sonography and MRI screening for claw hand deformity may offer important clues enabling early diagnosis of ML-III. PMID- 15361947 TI - Carbohydrate deficient glycoprotein syndrome type Ia. AB - Carbohydrate deficient glycoprotein syndromes (CDG) are inherited multisystem disorders characterized by the abnormal glycosylation of a number of serum glycoproteins. CDG-Ia results from deficiency of phosphomannomutase that catalyzes the conversion of mannose-6-phosphate to mannose-1-phosphate in the cytosol. We report a case of CDG-Ia in an 11-month-old girl with developmental delay, failure to thrive, inverted nipples and abnormal fat pads. The abnormal pattern of transferrin glycosylation and phosphomannomutase activity assay confirmed the diagnosis of CDG type Ia. Unfortunately, an efficient treatment is still not available for CDG type Ia patients. This is the first report of a Taiwanese patient with this syndrome. PMID- 15361948 TI - Hand-assisted laparoscopic surgery for abdominal aortic aneurysm. AB - There is a trend toward the use of minimally invasive surgery and limited incision for the surgical repair of abdominal aortic aneurysm (AAA). Conventional AAA repair is performed with a large laparotomy wound and uses either a transperitoneal or retroperitoneal approach. Due to the older age of this patient population, they often suffer more from the surgical wound, require prolonged hospital stay and have a slower recovery. We describe the use of hand-assisted laparoscopic surgery for grafting of a 5.5-cm infrarenal AAA identified by computed tomography scan. The operator inserted the left hand with a Pneumo Sleeve device in the abdomen through a 7-cm midline supraumbilical incision for laparoscopic surgery. Using the left hand and laparoscopic instruments to do the surgery provided control over the operation and sensation of touch. At the completion of the laparoscopic dissection, the first Clawford clamp was applied above the aortic bifurcation through the laparoscopic incision. The second Clawford clamp was applied below the renal arteries through the midline laparotomy incision. After cross-clamping the AAA, the aneurysm was incised, the clot removed, and lumbar bleeding points were oversewn with 3-O prolene suture. Then, the aneurysmal segment was grafted with a Dacron prosthesis using conventional suturing technique. The patient was extubated 3 hours after the operation, stayed in the intensive care unit for 1 night and was discharged 7 days later. He resumed oral intake on the postoperative day 1. There were no complications. This case illustrates that hand-assisted laparoscopic surgery for AAA can allow quick recovery of bowel function, quick progression to regular diet, short length of hospital stay, and probably a reduction in the total cost of care. PMID- 15361949 TI - Sensory neuropathy as the initial manifestation of multiple system atrophy. AB - Peripheral neuropathy is usually subtle or subclinical in patients with multiple system atrophy (MSA). A 54-year-old man who initially presented with sensory polyneuropathy developed extrapyramidal and cerebellar signs and autonomic failure typical of MSA 7 years later. Sural nerve biopsy disclosed a prominent loss of large myelinated fibers, whereas the unmyelinated fibers were totally spared. These findings imply that sensory neuropathy can be part of the multiple site degeneration in MSA, and that predominant involvement of large myelinated fibers of the sensory nerves can be a presenting feature. PMID- 15361950 TI - Difficulty in establishing sirolimus therapy as a cause of interstitial pneumonitis in renal-transplant recipients. PMID- 15361952 TI - [Longitudinal study and specific chemotherapy in children with chronic Chagas' disease, residing in a low endemicity area of Argentina]. AB - Clinical and epidemiological results of 95 treated and untreated chronic chagasic children, with an up to 24 years follow-up period are presented. This population studied in the 1/14 age bracket, residing in Santa Fe city, Argentina, was diagnosed through Chagas-specific conventional serologic reactions. Clinical examination was supplemented with electrocardiogram, chest X-rays, and blood and urine tests for evaluating hepatic function. The drugs employed were nifurtimox or benznidazole. In post treatment period xenodiagnosis was made in 33 patients. Regarding Trypanosoma cruzi transmission, the studied individuals presented multi risk antecedents: vectorial, congenital and/or blood transfusion. Among 24 untreated children 14 were controlled during 8/24 years: all this patients maintained the initial antibody concentration and clinical status. From 71 treated patients 49 were followed-up 4/24 years: 14 remained positive, 6 presented dubious results, and 29 showed final non-reactive results. 9 of this presented sometimes oscilating results. In 1/6 age bracket children, the serology turned negative after 3.5 years (median) once the treatment was finished, while patients treated in the 7/14 age bracket, the median of negativization was 8 years. 3.8% did not tolerate the drug. None of the groups changed their clinical condition. The untreated children did not change the serology. The percentage of treated children presenting negative serological results decrease according to the age when treatment was given: 75% became negative when treated at < or =4 years old and 43% when treated at > or =9 years old. PMID- 15361953 TI - [Exercise testing and 24 hours Holter monitoring in the detection of complex ventricular arrhythmias in different stages of chronic Chagas' heart disease]. AB - To detect complex ventricular arrhythmias in different stages of chronic chagasic cardiopathy, the results of exercise testing to 24 hours Holter monitoring have been compared. We evaluated a group of 71 patients, half women, aged 51+/-10.3, with no others associated diseases. These patients were separated in 4 groups according to degree of cardiac involvement. Statistical data can be found elsewhere in the study. In group IA, Holter monitoring detected 4.3% of complex ventricular arrhythmias, group IB 25%, group II 55% and group III 90%. We found no difference between Holter and exercise testing in the detection of complex ventricular arrhythmias in groups II and III (p = ns). In groups IA and IB we found 100% concordance, concerning the no detection of complex ventricular arrhythmias by exercise testing seen by two independent examiners. In group II there was a 70% concordance (kappa = 0.368, p = 0.003) and 90% in group III (kappa = 0.78, p = 0.002). Different results were found, concerning the presence of complex ventricular arrhythmias, among patients in the initial and advanced stages of chronic chagasic cardiopathy. In groups II and III we found no difference between the two methods in the detection of complex ventricular arrhythmias. It seems reasonable to recommend either Holter on exercise testing in groups IA and IB if progression of disease is noticed. PMID- 15361954 TI - [Erythema nodosum leprosum case series report: clinical profile, immunological basis and treatment implemented in health services]. AB - Erythema nodosum leprosum is an acute inflammatory event in the chronic course of leprosy. It is considered an immunological disorder and an important cause of morbidity and disability. We evaluate the clinical profile, serology and histopathology 58 erythema nodosum leprosum patients sequentially recruited, from July- December 2000, in an endemic area in Central Brazil (Goias State). Half of the reactins were considered severe and 66% of the cases had the first episode of reaction during specific treatment. The majority of patients and controls were positive to anti-PGL-I IgM. The more frequent histopathological findings in erythema nodosum leprosum were presence of intracellular acid-fast bacilli, perivascular/peradnexial mononuclear inflammatory infiltrate, and neural aggression. Ninety six percent of the patients were treated with systemic steroid in the first episode. The results point out to the association between ENL and neuritis and the rare adoption of thalidomide as a solely medication in the health services. PMID- 15361955 TI - [HIV and hepatitis B virus co-infection: prevalence and risk factors]. AB - The objective of this study was to assess the prevalence of hepatitis B virus and possible risk factors for this disease in 401 patients infected with the human immunodeficiency virus, followed at the University Hospital of the Ribeirao Preto Medical School, Sao Paulo State University. Each participant was submitted to a specific questionnaire and had a blood sample tested for the serologic markers HBsAg, total anti-HBcAg, anti-HBsAg and anti-HCV, using ELISA technique. The overall prevalence of hepatitis B markers was 40.9%, with 8.5% for HBsAg, 39.7% for total anti-HBcAg and 5.5% for anti-HBsAg. The variables that showed association with HBV were: age, higher education level, history of jaundice, time spent in prison, having a homosexual partner and positive markers for anti-HCV. Co-infection HBV/HCV was present in 20.4% of the participants. PMID- 15361956 TI - [Larvicidal activity of tannins isolated of Magonia pubescens St. Hil. (Sapindaceae) against Aedes aegypti (Diptera, Culicidae)]. AB - Phytochemical study of the larvicidal fractions which were carried out for the first time, isolated of the Magonia pubescens, monitored by the study of efficacy against the 3rd larval instar of Aedes aegypti, in the search of alternatives for the control of that mosquito and to obtain structures susceptible to chemical improving of the activity for the synthetic via of other derived. The fractions with biological activity were monitored chemically through chromatography of thin layer, using as revealing a solution acid of vanillin, analyzed by nuclear magnetic resonance of hydrogen and spectrometry of masses. The bioassays with the fractions were accomplished with five replications, controlled at the temperature of 28+/-1 degrees C, 80+/-5% of relative humidity and 12 h light. The found lethal concentrations of the fraction that it presented the largest potential larvicidal, MP-9, LC50 and LC90, were of 3.1 and 36.6 ppm, respectively. All the experiments were accompanied by a control series, containing the same number of larvae. PMID- 15361957 TI - [Evaluation of the impact on Aedes aegypti infestation in cement tanks of the municipal district of Caninde, Ceara, Brazil after using the Betta splendens fish as an alternative biological control]. AB - Caninde has a population of 71,235 inhabitants. In April 2001, the city started using larvivorous fish in cement water tanks, as a means of biological control of Aedes aegypti larvae. During house-to-house visits by health agents, instead of treating the water tank with larvicide, a Betta splendens fish was introduced into each tank. The number of houses and the number of tanks was estimated by monthly surveys. Then, the number of tanks per house was determined. Taking into account this estimated number and the number of houses visited, the infestation level for each kind of deposit was analyzed. In January 2001, 70.4% of the water tanks presented mosquitoes. Following the intervention, in January 2002 only 7.4% were positive and by December 2002 the rate had dropped to 0.2%. The efficacy of Betta splendens as a biological control agent in cement water tanks was clearly demonstrated, by achieving a 320 times reduction in the infestation level. PMID- 15361958 TI - Training-related accidents during teacher-student-assistance activities of medical students. AB - A survey was done to determine the most common hospital accidents with biologically contaminated material among students at the Medical College of the Federal University of Minas Gerais. Six hundred and ninety-four students (between fifth and twelfth semesters of the college course) answered the questionnaire individually. Three-hundred and forty-nine accidents were reported. The accident rate was found to be 33.9% in the third semester of the course, and increased over time, reaching 52.3% in the last semester. Sixty-three percent of the accidents were needlestick or sharp object injuries; 18.3% mucous membrane exposure; 16.6% were on the skin, and 1.7% were simultaneously on the skin and mucous membrane exposure. The contaminating substances were: blood (88.3%), vaginal secretion (1.7%), and others (9.1%). The parts of the body most frequently affected were: hands (67%), eyes (18.9%), mouth (1.7%), and others (6.3%). The procedures being performed when the accidents occurred were: suture (34.1%), applying anesthesia (16.6%), assisting surgery (8.9%), disposing of needles (8.6%), assisting delivery (6.3%), and others (25.9%). Forty-nine percent of those involved reported the accident to the accident control department. Of these 29.2% did not receive adequate medical assistance. Eight percent of those involved used antiretroviral drugs and of these 86% discontinued the treatment on receiving the Elisa method applied to the patient (HIV-negative); 6.4% discontinued the treatment due to its side-effects; and 16% completed the treatment. PMID- 15361959 TI - Prevalence of Epstein-Barr virus antibodies in healthy children and adolescents in Vitoria, State of Espirito Santo, Brazil. AB - The prevalence and age distribution of Epstein-Barr virus infection varies in different populations and there is little information about the epidemiology of this infection in Brazil. We studied the prevalence of EBV antibodies in a sample of 283 children and adolescents between 1 and 21 years old. The sample was taken from two neighborhoods in Vitoria (capital city of Espirito Santo, Brazil). The Sao Pedro (SP) neighborhood represented an area with lower socioeconomic status and the Praias (P) neighborhood represented an area with higher SES. Anti-VCA (Virus Capsid Antigen) antibodies were detected by ELISA and anti-EBNA (Epstein Barr Nuclear Antigen) antibodies were detected by an anti-complement immunofluorescence method, both using commercial kits. The results showed an overall prevalence rates of anti-VCA and anti-EBNA of 71% and 54% respectively. The prevalence for both anti-EBV antibodies was higher and probably the infection occurred earlier in the SP neighborhood. Among the various socioeconomic factors studied only low family income and maternal education level were significantly correlated with a higher frequency of positive serology for anti-VCA. These results demonstrate that there is a high prevalence of EBV antibodies in children and adolescents living in Vitoria, that occurs more frequently at a younger age in children from families with low socioeconomic status. In addition, the results demonstrate an intermediate age distribution pattern between those reported in developed and underdeveloped countries. PMID- 15361960 TI - [Endemic dengue: surveillance strategy challenges]. AB - Interaction between clinical infection, vector, and population serotype immunity defines dengue epidemic risk. An endemic-sporadic epidemiological situation is an acceptable control target in metropolitan regions deficient in urban upgrading, sanitation, and health agents' access to residences, besides low residual power of biological insecticides and social mobilization problems. Surveillance requires continued action by government and society (particularly when incidence decreases) and adequate proposals. To establish a sensitive and specific surveillance system for endemic periods, the authors propose the sentinel clinical component as part of emergency care (detecting serious tip-of-the iceberg cases) and reference services for acute febrile diseases, defining clinical forms and providing diagnostic confirmation. Although complex, sample serotype surveys should be conducted in strategic areas, evaluating: immunity and susceptibility of age groups to circulating serotypes; estimation of sub-clinical cases; and Surveillance and Control System reach. PMID- 15361961 TI - [Fascioliasis: report of two cases from rural areas of Rio de Janeiro]. AB - We describe two cases of fascioliasis from rural areas of Rio de Janeiro that are endemic for schistosomiasis, both of which were found during a coprological survey. The patient from Paracambi complained of dizziness. The patient from Sumidouro complained of dizziness, tiredness and cough. She was treated with praziquantel and her stool parasitological examination became negative. PMID- 15361962 TI - [Lonomia erucism in Teresopolis, Rio de Janeiro State, Brazil: report of a probable case and review]. AB - This is a case report of a 44-year-old male living in Teresopolis, RJ, Brazil, probably poisoned by contact with a Lonomia caterpillar, who presented hemolytic anemia, decreased platelet count and acute renal insufficiency. Lonomia erucism diagnosis was established by anamnesis and clinical and laboratory manifestations. Therapeutic measures consisting of hemotransfusion and hemodialysis were successful. Physiopathologic and clinical features of erucism by Lonomia are discussed. PMID- 15361963 TI - [Intestinal parasites in nursery schools of Lages, southern Brazil]. AB - Two hundred children from nursery schools in Lages, southern Brazil, were associated as to parasitic infections. The overall prevalence of helminths and protozoa was 70.5%, affecting 61.4% of male and 74.5% of female children. The most prevalent parasites were Ascaris lumbricoides (35%), Giardia lamblia (14%) e Trichuris trichiura (13%). PMID- 15361964 TI - [Erucism]. PMID- 15361965 TI - Paracoccidioidomycosis: an unusual presentation with a rapid progression. PMID- 15361966 TI - [Frederico Adolfo Simoes Barbosa (1916-2004)]. PMID- 15361967 TI - Vibrio spp. and Salmonella spp., presence and susceptibility in crabs Ucides cordatus. AB - The presence of Vibrio spp. and Salmonella spp. in crabs marketed at the Bezerra de Menezes Ave., Fortaleza, State of Ceara, Brazil, was assessed between February and May, 2003. The number of individuals sampled in each one of the fifteen weekly samplings ranged between four and eight. Seven strains of Salmonella, from four different samplings, were identified, being five of them identified as serotype S. Senftenberg and two as S. Poona. All strains of Salmonella were sensitive to the tested anti-microbial drugs, with the exception of tetracycline and nalidixic acid, for which an intermediary sensibility was found. The MPN's for Vibrio ranged between 110/g and 110,000/g. Of the forty five Vibrio strains isolated from the crab samples, only 10 were identified up to the species level: two V. alginolyticus and eight V. parahaemolyticus. Bacteria belonging to the Enterobacteriaceae and Pseudomonaceae families were also identified, namely Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pantoea agglomerans and Pseudomonas aeruginosa. The proper cooking of the animals is recommended in order to avoid problems for the consumers of this crustacean. PMID- 15361968 TI - PCR -- based diagnosis to evaluate the performance of malaria reference centers. AB - Although the Giemsa-stained thick blood smear (GTS) remains the gold standard for the diagnosis of malaria, molecular methods are more sensitive and specific to detect parasites and can be used at reference centers to evaluate the performance of microscopy. The description of the Plasmodium falciparum, P. vivax, P. malariae and P. ovale ssrRNA gene sequences allowed the development of a polymerase chain reaction (PCR) that had been used to differentiate the four species. The objective of this study was to determine Plasmodium species through PCR in 190 positive smears from patients in order to verify the quality of diagnosis at SUCEN's Malaria Laboratory. Considering only the 131 positive results in both techniques, GTS detected 4.6% of mixed and 3.1% of P. malariae infections whereas PCR identified 19.1% and 13.8%, respectively. PMID- 15361969 TI - A morphological protocol and guide-list on uterine cervix cytology associated to Papillomavirus infection. AB - The present study was designed to further assess the validity of the cytological description of morphological lesions said to be related to Papillomavirus (HPV) infections in senior women. The casuistic comprised 196 cervical smears from a group of women with no clinical or morphological evidence of neoplasia, collected simultaneously with samples submitted to detection of HPV DNA by PCR in a previous study. Three experienced cytologists studied each slide in two different conditions, with an interval of 20 months between them. The first approach was performed under routine laboratory standards, whereas the second was guided by a list of 16 well-defined parameters indicative of HPV-related cytological lesions. When suspicious cases of HPV-related alterations were grouped with positive cases, they showed on average: sensitivity of 25.5%, specificity of 84.4% and positive predictive value (PPV) of 26.8%. When suspicious cases were grouped with negative cases, sensitivity decreased, whereas specificity and PPV increased, as expected. In the second reading, which followed a "guide-list", a decrease in sensitivity was observed, contrasting with a sharp increase of positive predictive value. Among the 16 cytomorphological criteria tested, "koilocytosis", "mild koilocytosis" and "condylomatous parabasal cells" yielded the best predictive value for HPV DNA detection by PCR. In conclusion, despite the low sensitivity, cytopathologic assessment of cervico-vaginal smears leads to a highly specific diagnosis of HPV infection in menopausal women, with PPV of 91.0% when directed by a guide-list of well-defined morphologic criteria. PMID- 15361970 TI - Oligodendroglioma in a patient with AIDS: case report and review of the literature. AB - In the last years, new techniques of neuroimages and histopathological methods have been added to the management of cerebral mass lesions in patients with AIDS. Stereotactic biopsy is necessary when after 14 days of empirical treatment for Toxoplasma gondii encephalitis there is no clinical or neuroradiologic improvement. We report a woman with AIDS who developed a single focal brain lesion on the right frontal lobe. She presented a long history of headache and seizures. After two weeks of empirical treatment for toxoplasma encephalitis without response, a magnetic resonance image with spectroscopy was performed and showed a tumoral pattern with a choline peak, diminished of N-acetyl-aspartate and presence of lactate. A stereotactic biopsy was performed. Histopathological diagnosis was a diffuse oligodendroglioma type A. A microsurgical resection of the tumor was carried out and antiretroviral treatment was started. To date she is in good clinical condition, with undetectable plasma viral load and CD4 T cell count > 200 cell/uL. PMID- 15361971 TI - Chagasic meningoencephalitis: case report of a recently included AIDS-defining illness in Brazil. AB - Recently, reactivation of Chagas disease (meningoencephalitis and/or myocarditis) was included in the list of AIDS-defining illnesses in Brazil. We report a case of a 52-year-old patient with no history of previous disease who presented acute meningoencephalitis. Direct examination of blood and cerebrospinal fluid (CSF) showed Trypanosoma cruzi. CSF culture confirmed the diagnosis. Serological assays for T. cruzi and human immunodeficiency virus (HIV) were positive. Despite treatment with benznidazol and supportive measures, the patient died 24 hours after hospital admission. In endemic areas, reactivation of Chagas disease should always be considered in the differential diagnosis of meningoencephalitis among HIV-infected patients, and its presence is indicative of AIDS. PMID- 15361972 TI - Neurocryptococcosis: diagnosis by PCR method. AB - Cryptococcus neoformans detection was optimized using PCR technique with the objective of application in the clinical laboratory diagnosis. The amplification area was ITS and 5,6S which encodes the ribosomal RNA (rRNA). A total of 72 cerebrospinal fluid (CSF) samples were used, obtained from cases with and without AIDS. The patients had cryptococcal meningitis (n = 56) and meningitis caused by other agents (n = 16). The results demonstrated that PCR test had the highest sensitivity rates, superior to culture (85.7%) and to India ink test (76.8%). PCR was found to be sensitive in detecting 1 cell/mL and highly specific since it did not amplify other fungal DNA. The comparative analysis of the methods showed that PCR is more sensitive and specific and is applicable as an important laboratorial resource for neurocryptococcosis diagnosis. PMID- 15361973 TI - The perception, beliefs and practices toward genitourinary schistosomiasis by inhabitants of selected endemic areas (Edo/Delta States) in south-eastern Nigeria. AB - Well-structured questionnaire on the perception, impression and response to genitourinary bilharziasis (Genitourinary schistosomiasis) was administered and explained in local languages: 'Igbo' 'Esan' 'Ezon' Itshekiri and Bini to 33815 inhabitants of selected endemic areas in south-eastern Nigeria from January, 1999 to December, 2001. Out of this number, 3815 (11.3%) were properly filled and returned. About 42.0% of the inhabitants admitted knowledge of the disease, while 14 (0.4%) knew about the aetiologic agent. About 181 (5.0%) who responded, admitted procuring treatment, while 100 (5.0%) declined to seek treatment of any sort. The relationships between water-bodies and human activities, and infection were well discussed. Amongst those who admitted knowledge of the disease but no knowledge of its etiologic agent, declined seeking treatment of any kind, but believe the disease is a natural phenomenon in ones developmental stage and therefore of no morbidity and mortality. Laboratory analysis of urine, faeces, semen and HVS was employed to assess questionnaire responses, and in some cases, physical examination was utilized to augment laboratory analysis in confirming urinal diagnosis. Haematuria was only directly related to egg count in the early part of life. Females were significantly haematuric and excreted more ova than males (p < 0.05). Headache (43.0%) and fever (31.0%) were major clinical signs while sexual pains (22.0%) were the least. PMID- 15361974 TI - Eco-epidemiological aspects of Trypanosoma cruzi, Trypanosoma rangeli and their vector (Rhodnius pallescens) in Panama. AB - The eco-epidemiology of T. cruzi infection was investigated in the Eastern border of the Panama Canal in Central Panama. Between 1999 and 2000, 1110 triatomines were collected: 1050 triatomines (94.6%) from palm trees, 27 (2.4%) from periurban habitats and 33 (3.0%) inside houses. All specimens were identified as R. pallescens. There was no evidence of vector domiciliation. Salivary glands from 380 R. pallescens revealed a trypanosome natural infection rate of 7.6%, while rectal ampoule content from 373 triatomines was 45%. Isoenzyme profiles on isolated trypanosomes demonstrated that 85.4% (n = 88) were T. cruzi and 14.6% (n = 15) were T. rangeli. Blood meal analysis from 829 R. pallescens demonstrated a zoophilic vector behavior, with opossums as the preferential blood source. Seroprevalence in human samples from both study sites was less than 2%. Our results demonstrate that T. cruzi survives in the area in balanced association with R. pallescens, and with several different species of mammals in their natural niches. However, the area is an imminent risk of infection for its population, consequently it is important to implement a community educational program regarding disease knowledge and control measures. PMID- 15361975 TI - Studies on entomological monitoring: mosquito species frequency in riverine habitats of the Igarapava Dam, Southern Region, Brazil. AB - Diversity of mosquito species was evaluated in different habitats before and after the Igarapava reservoir flooding in the Grande River, Southern Cerrado of Brazil. We aimed at verifying changes in these mosquito populations in consequence of the lake formation. Four habitats were selected as sampling stations: peridomiciliary habitat, pasture, "veredas" and gallery forest patch. Bimonthly collections were made with the Shannon trap and human bait, including diurnal, crepuscular and nocturnal period of mosquito activity. The Shannon Index results from the potential vectors were compared using Student t-test. Aedes scapularis, Anopheles darlingi and An. albitarsis senso latu seasonal abundance were described with moving average and compared using chi2 test. There were changes in the mosquito frequency in the habitats, except for the "veredas" that was 13 km away from the catchment area. The altering in mosquito species seasonal abundance suggests breeding places expansion. Diversity indexes can be used to monitor changes in mosquito vector population in environments where abrupt disturbance can alter disease transmission cycles. PMID- 15361976 TI - Synergistic action of praziquantel and host specific immune response against Schistosoma mansoni at different phases of infection. AB - The interaction between specific immune response to Schistosoma mansoni and praziquantel (PZQ) was studied in mice. In mice harboring concomitant immunity, 6 day-old parasites treated with PZQ were more effectively removed than 24 h treated parasites despite both had a significant worm burden reduction when compared with respective treated controls. These results show that PZQ can be effective at the skin and lung stages of parasite's development mainly acting with a established specific immune response, and particularly at the lung phase. PMID- 15361977 TI - Relationship of cryptosporidiosis to abdominal pain and diarrhea in Mayan Indians. AB - Demonstration of cryptosporidiosis in Mayan Indians living around Lake Atitlan provided an opportunity to correlate infection with abdominal pain and/or diarrhea in different age groups of children. 94 subjects experiencing abdominal pain and/or diarrhea, between the ages of 2 and 13 were studied in towns around Lake Atitlan, Guatemala, over a two-year period. Cryptosporidium oocysts were found in the feces of 29% of children who presented with abdominal pain and 21% with diarrhea. Of the 60 infected subjects, 45% were experiencing abdominal pain and 33% diarrhea, 22% had abdominal pain and diarrhea. Both abdominal pain and diarrhea were significantly higher in children under 10 years of age and were most prevalent in the 6-9 year old age group but the correlation of symptoms to infection was not significantly different as the ages of the children increased. The high frequency of abdominal pain and/or diarrhea with infection in children was consistent with cryptosporidiosis, a disease considered as one of several common intestinal infections that produce these symptoms. PMID- 15361978 TI - Causes of low vision and use of optical aids in the elderly. AB - PURPOSE: To determine the causes of low vision in an elderly population attended by a university visual rehabilitation service and to check for the use of prescribed optical aids. METHOD: A cross-sectional study was carried out on patients aged 60 years or over attending for the first time a university low vision service in 2001. Ophthalmic reevaluation and interview were performed by means of a structured questionnaire in 2002. RESULTS: The sample comprised 50 subjects aged between 60 and 90 years. Severe low vision (< or =20/200) was present in 68.0% of patients. The main cause of low vision was age-related macular degeneration (44.0%). Regarding literacy, 16.0% were illiterate and 72.0% had completed fundamental schooling. Thirty-one patients (62.0%) had been prescribed optical aids; 54.8% of these patients stated that they use them. A majority (70.6%) held a favorable opinion of these aids. CONCLUSIONS: The main cause of low vision was age-related macular degeneration. Approximately half of those receiving prescriptions reported actually using the aids in their daily activities. Making best use of residual vision in the elderly population with visual impairment is a priority, given the social context, if the independence necessary for enhanced quality of life is to be achieved. PMID- 15361979 TI - Subjective versus objective stress in noncritically ill hospitalized and outpatient adult men. AB - A cross-sectional study of 120 subjects was performed with the purpose of evaluating stress hormones and emotional stress (anxiety) in outpatient and hospitalized subjects. The aims were to determine the degree of objective stress, as well as to correlate this finding with subjective findings, estimated using Beck's Anxiety Inventory. METHOD: Three populations were investigated, namely outpatient clinical cases (Group I, n = 30), hospitalized clinical individuals (Group II, n = 30), and hospitalized surgical candidates (Group III, n = 30). Controls (Group IV, n = 30) were healthy volunteers who were health-care professionals and students. To avoid hormone interactions, only men were enrolled in all groups. All hospitalized subjects were tested on admission and before therapeutic interventions. Fasting epinephrine, norepinephrine, and cortisol were measured in the morning, and Beck's Anxiety Inventory was adminstered by a trained psychologist. RESULTS: The 3 patient groups displayed higher anxiety levels than the controls. Hormone concentrations did not present remarkable changes and did not correlate with subjective stress (anxiety). CONCLUSIONS: 1) Subjective disorders (as determined with Beck's Anxiety Inventory ) were a common finding in both outpatient and hospitalized populations, without differences between the various groups; 2) Objective stress (as determined by elevated hormone levels) was more difficult to confirm--findings rarely exceeded the reference range; 3) Correlation between the two variables could not be demonstrated; 4) Further studies are necessary to define stress quantification and interpretation in patient populations, especially in relationship with nutritional diagnosis and dietetic prescription. PMID- 15361980 TI - Surgical treatment of rectal prolapse: experience and late results with 51 patients. AB - The "best" surgical technique for the management of complete rectal prolapse remains unknown. Due to its low incidence, it is very difficult to achieve a representative number of cases, and there are no large prospective randomized trials to attest to the superiority of one operation over another. PURPOSE: Analyze the results of surgical treatment of complete rectal prolapse during 1980 and 2002. METHOD: Retrospective study. RESULTS: Fifty-one patients underwent surgical treatment during this period. The mean age was 56.7 years, with 39 females. Besides the prolapse itself, 33 patients complained of mucous discharge, 31 of fecal incontinence, 14 of constipation, 17 of rectal bleeding, and 3 of urinary incontinence. Abdominal operations were performed in 36 (71%) cases. Presacral rectopexy was the most common abdominal procedure (29 cases) followed by presacral rectopexy associated with sigmoidectomy (5 cases). The most common perineal procedure was perineal rectosigmoidectomy associated with levatorplasty (12 cases). Intraoperative bleeding from the presacral space developed in 2 cases, and a rectovaginal fistula occurred in another patient after a perineal rectosigmoidectomy. There were 2 recurrences after a mean follow-up of 49 months, which were treated by reoperation. CONCLUSION: Abdominal and perineal procedures can be used to manage complete rectal prolapse with safety and good long-term results. Age, associated medical conditions, and symptoms of fecal incontinence or constipation are the main features that one should bear in mind in order to choose the best surgical approach. PMID- 15361981 TI - P53 and Rb tumor suppressor gene alterations in gastric cancer. AB - Inactivation of tumor suppressor genes has been frequently observed in gastric carcinogenesis. Our purpose was to study the involvement of p53, APC, DCC, and Rb genes in gastric carcinoma. METHOD: Loss of heterozygosity of the p53, APC, DCC and Rb genes was studied in 22 gastric cancer tissues using polymerase chain reaction; single-strand conformation polymorphism of the p53 gene exons 5-6 and exons 7-8 was studied using 35S-dATP, and p53 expression was detected using a histological immunoperoxidase method with an anti-p53 clone. RESULTS AND DISCUSSION: No loss of heterozygosity was observed in any of these tumor suppressor genes; homozygous deletion was detected in the Rb gene in 23% (3/13) of the cases of intestinal-type gastric carcinoma. Eighteen (81.8%) cases showed band mobility shifts in exons 5-6 and/or 7-8 of the p53 gene. The presence of the p53 protein was positive in gastric cancer cells in 14 cases (63.6%). Normal gastric mucosa showed negative staining for p53; thus, the immunoreactivity was likely to represent mutant forms. The correlation of band mobility shift and the immunoreactivity to anti-p53 was not significant (P =.90). There was no correlation of gene alterations with the disease severity. CONCLUSIONS: The inactivation of Rb and p53 genes is involved in gastric carcinogenesis in our environment. Loss of the Rb gene observed only in the intestinal-type gastric cancer should be further evaluated in association with Helicobacter pylori infection. The p53 gene was affected in both intestinal and diffuse histological types of gastric cancer. PMID- 15361982 TI - Pulmonary function and aerobic capacity in asymptomatic bariatric candidates with very severe morbid obesity. AB - PURPOSE: Aerobic capacity and respiratory function may be compromised in obesity, but few studies have been done in highly obese bariatric candidates. In a prospective study, these variables were documented in the preoperative period, aiming to define possible physiologic limitations in a apparently healthy and asymptomatic population. METHOD: Forty-six consecutively enrolled adults (age 39.6 +/- 8.4 years, 87.0% females, body mass index /BMI 49.6 +/- 6.3 kg/m2) were analyzed. Ventilatory variables were investigated by automated spirometry, aerobic capacity was estimated by a modified Bruce test in an ergometric treadmill, and body composition was determined by bioimpedance analysis. RESULTS: Total fat was greatly increased (46.4 +/- 4.6% of body weight) and body water reduced (47.3 +/- 4.6 % body weight), as expected for such obese group. Spirometric findings including forced vital capacity of 3.3 +/- 0.8 L and forced expiratory volume-1 second of 2.6 +/- 0.6 L were usually acceptable for age and gender, but mild restrictive pulmonary insufficiency was diagnosed in 20.9%. Aerobic capacity was more markedly diminished, as reflected by very modest maximal time (4.5 +/- 1.1 min) and distance (322 +/-142 m) along with proportionally elevated maximal oxygen consumption (23.4 +/- 9.5 mL/kg/min) achieved by these subjects during test exercise. CONCLUSIONS: 1) Cardiopulmonary evaluation was feasible and well-tolerated in this severely obese population; 2) Mean spirometric variables were not diminished in this study, but part of the population displayed mild restrictive changes; 3) Exercise tolerance was very negatively influenced by obesity, resulting in reduced endurance and excessive metabolic cost for the treadmill run; 4) More attention to fitness and aerobic capacity is recommended for seriously obese bariatric candidates; PMID- 15361983 TI - Impact of alcohol intoxication and withdrawal syndrome on social phobia and panic disorder in alcoholic inpatients. AB - PURPOSE: To investigate the impact of alcohol intoxication and withdrawal on the course of social phobia and panic disorder. METHOD: A group of 41 alcoholic inpatients undergoing detoxification therapy were interviewed using the SCID-I (DSM-IV) and questions to detect fluctuations in the course of social phobia and panic disorder as a function of the different phases in alcohol dependence (intoxication, withdrawal, and lucid interval). RESULTS: Only 1 (2.4%) patient presented panic disorder throughout life, and 9 (21.9%) had panic attacks during alcohol intoxication or during the withdrawal syndrome. Sixteen (39%) alcoholic patients showed social phobia with onset prior to drug use. However, drinking eventually became unable to alleviate social phobia symptoms or worsened such symptoms in 31.2% of social-phobic patients. While patients with social phobia reported a significant improvement in psychiatric symptoms during alcohol intoxication, patients experiencing panic attacks worsened significantly during intoxication. In the withdrawal phase, patients with social phobia tended to have more and more intense phobic symptoms. CONCLUSION: Our findings indicate that the impact of alcohol intoxication is different for social phobia as compared to panic disorder, at first decreasing the social-phobic symptoms but later aggravating them. In panic disorder, the impact of intoxication by alcohol is more harmful, at least in the short term. PMID- 15361984 TI - The effect of the presence of muscle tissue in a bone healing site. AB - PURPOSE: The recovery of a bone fracture is a process that is not yet fully understood. The literature conflicts on the results obtained by the interposition of foreign tissue inside a damaged bone. The objective of the present study was to ascertain the effect of placing muscle tissue between the stumps of a fractured bone. METHOD: The study was carried out on 10 rabbits divided into 2 groups (n = 5): Group 1--partial fracture of the humerus and interposition of muscle tissue; Group 2--complete fracture of the humerus and interposition of muscle tissue. The fractured limb of all animals was immobilized for 8 weeks. At the end of this time, the rabbits were killed and their operated humeri were carefully removed for roentgenological and histological assessment. RESULTS: All humeri of Group 1 recovered their integrity and normal aspect. However, the healing of the humeri of Group 2 was not perfect. Gross angulation of the bone diaphysis occurred in all animals, and immature trabecular bone, osteochondral tissue, and persistence of muscle tissue substituted normal bone. CONCLUSIONS: Interposed muscle does not affect partial bone fracture healing but causes instability in a complete fracture. PMID- 15361985 TI - Umbilical mass as the sole presenting symptom of pancreatic cancer: a case report. AB - Umbilical nodes are rare. The metastatic involvement of the region was first described in 1846. Sister Mary Joseph was the first observer to establish the correlation between carcinomas and umbilical nodes. The umbilical node may be the sole presenting sign of cancer and is usually associated with advanced disease and poor prognosis. A 64-year-old woman, previously healthy, presented vague abdominal discomfort and a hard umbilical nodule for 1 week, which was first diagnosed as an incarcerated umbilical hernia. She underwent a new clinical assessment and biopsy. After immunohistochemical analysis and computerized tomography, she was diagnosed with pancreatic cancer. The clinical staging showed advanced disease with distant metastasis. She received palliative chemotherapy. After 8 months, she was alive in poor clinical condition. Clinical suspicion should lead to a careful additional evaluation whenever an umbilical nodule presents with malignant signs. PMID- 15361986 TI - Finasteride-associated male infertility. AB - Finasteride is a potent and specific inhibitor of the 5alpha-reductase enzyme in men. Clinical studies have shown that finasteride 1mg/day is effective for promoting hair growth in men with male pattern hair loss. However, there is a concern about the use of finasteride, especially in young fertile patients, because of its action on testosterone metabolism. This paper describes 3 cases of young patients who had very poor seminal quality during finasteride treatment (1 mg/day), and their seminal quality greatly improved after cessation of finasteride treatment. Two of them presented with a left varicocele and the other was obese. We hypothesize that finasteride may not dramatically change the spermatogenesis process in healthy men, but in patients with conditions related to infertility, an amplification of the negative influence of finasteride could occur. Future studies should be done to clarify the extent of the effect of finasteride in patients fertility problems. PMID- 15361987 TI - Voiding dysfunction and urodynamic abnormalities in elderly patients. AB - Lower urinary tract dysfunction is a major cause of morbidity and decreased quality of life in elderly men and women. With the progressive aging of the population, it is important to understand common micturitional disorders that may occur in this population. Most urinary problems in the elderly are multifactorial in origin, demanding a comprehensive assessment of the lower urinary tract organs, functional impairments, and concurrent medical diseases. Urodynamics is a highly valuable tool in the investigation of elderly patients with lower urinary tract symptoms. Urodynamic tests are not always necessary, being indicated after excluding potentially reversible conditions outside the urinary tract that may be causing or contributing to the symptoms. Although urodynamic tests may reveal common diagnoses such as bladder outlet obstruction and stress urinary incontinence in the elderly population, findings such as detrusor overactivity and impaired detrusor contractility are common and have important prognostic and therapeutic implications. The purpose of this article is to describe common urologic problems in the elderly and review the indications for and clinical aspects of urodynamic studies in these conditions. PMID- 15361988 TI - Hepatitis B: epidemiological, immunological, and serological considerations emphasizing mutation. AB - The global prevalence of hepatitis B virus is estimated to be 350 million chronic carriers, varying widely from low (<2%, as in Western Europe, North America, New Zealand, Australia, and Japan) to high (>8% as in Africa, Southeast Asia, and China). The overall prevalence in Brazil is about 8%. There are currently 7 genotypic variations, from A to G, and also 4 main surface antigen subtypes: adw, ayw, adr, and ayr. There has been great interest in identifying the geographic distribution and prognosis associated with the various genotypes and subtypes. Although the serologic test is highly sensitive and specific, it does not detect cases of mutant hepatitis B, which is increasingly common worldwide due to resistance and vaccine escape, antiviral therapy, and immunosuppression, among other causes. Alterations in surface, polymerase, X region, core, and precore genes have been described. The main mutations occur in surface and in core/precore genes, also known as occult hepatitis, since its serologic markers of active infection (HBsAg) and viral replication (HBeAg) can be negative. Thus, mutation should be suspected when serologic tests to hepatitis B show control of immunity or replication coincident with worsened clinical status and exclusion of other causes of hepatitis. PMID- 15361989 TI - Surgical infections: a microbiological study. AB - Surgical infections are mostly polymicrobial, involving both aerobes and anaerobes. One hundred seventeen cases comprised of abscesses (n=51), secondary peritonitis (n=25), necrotizing fascitis (n=22) and wounds with devitalized tissues (n=19) were studied. The number of microorganisms isolated per lesion was highest in secondary peritonitis (2.32). The aerobe/ anaerobe ratio was 0.81 in secondary peritonitis and 1.8 in necrotizing fascitis. Most secondary peritonitis (80%), necrotizing fascitis (75%) and wounds with devitalized tissues (66.7%) were polymicrobial. Common microorganisms isolated in our study were E. coli, Staphylococcus aureus, Klebsiella spp., Pseudomonas aeruginosa, Bacteroides fragilis and Peptostreptococcus spp. The most effective antibiotics for S. aureus were clindamycin (79.1%) and cefuroxime (70.8%). For Gram-negatives (Klebsiella spp., E. coli and Proteus spp.), the most effective antibiotics were cefotaxime, ceftizoxime, amikacin and ciprofloxacin. Pseudomonas aeruginosa was maximally sensitive to amikacin (35.2%) and ciprofloxacin (35.2%). The greatest degree of multidrug resistance to all the drugs was found in P. aeruginosa (52.9%), followed by Klebsiella spp. (33.3%), Proteus spp. (33.3%), E. coli (22.2%), and S. aureus (12.5%). All the anaerobes that we isolated were 100% sensitive to metronidazole and chloramphenicol, followed by clindamycin (95% to 100%). Apart from antibiotic therapy, non-antimicrobial methods, such as hyperbaric oxygen therapy and debridement also play an important role in the treatment of surgical infections. PMID- 15361990 TI - CD81 binding regions of hepatitis C virus remain conserved after liver transplantation. AB - CD81 is a surface-associated protein expressed in the membranes of mammalian cells. It has been suggested that CD81 interacts with hepatitis C virus E2 protein, and thus might facilitate the entry of HCV into hepatocytes. The envelope-binding site appears to involve amino acids (aa) 480-493 and 544-551 within the E2 glycoprotein. Little is known about the quasispecies genetic diversity of these two regions. We studied four patients who underwent transplantation for HCV-related cirrhosis and who developed recurrent hepatitis C. We evaluated HCV quasispecies diversity in serum samples obtained at the time of transplantation and at several time points thereafter. Quasispecies diversity was assessed by cloning and sequencing of viral isolates, with computer analysis of evolution models. The genetic distance in the region that spans aa 480 to 493 was 0.019 +/- 0.004 before the transplant, and 0.039 +/- 0.014 after the transplant (p=0.324). In the aa 544 to 551 region, the pre-transplant genetic distance was 0.012 +/- 0.008 and the post-transplant distance, 0.010 +/- 0.007 (p=0.890). There was also no significant difference between the number of nonsynonymous substitutions per nonsynonymous site before and after transplantation. In conclusion, the HCV genetic sequences of putative CD81 binding regions aa 480-493 and aa 544-551 did not diversify significantly after liver transplantation. This may favor HCV re-infection of the allograft after liver transplantation. PMID- 15361991 TI - Seroprevalence of hepatitis B and C in the Western Brazilian Amazon region (Rio Branco, Acre): a pilot study carried out during a hepatitis B vaccination program. AB - In 1999, on the occasion of the application of the first vaccine dose during the state vaccination campaign against hepatitis B virus (HBV), 390 individuals from the town of Rio Branco, Acre, aged two or more years were selected for the determination of the seroprevalence of HBV and HCV. HBV markers (HBsAg, anti-HBs, and anti-HBc IgG) were determined on this occasion and anti-HBs antibodies were also assessed 30 days after the third vaccine dose. At the time of vaccination, 39% of the individuals were still susceptible to HBV, while 61% presented serologic evidence of previous HBV contact or previous vaccination. The individuals with previous HBV contact were significantly older (p<0.001) than those without HBV markers. Of the 192 individuals who returned for reexamination, 30 days after the third dose, 158 (82.3%) had received three vaccine doses, and only 60 (31.2%) belonged to the group without HBV markers. In these individuals, the seroconversion rate after the third dose was 92% (55/60). In conclusion, we found considerable HBV in this population, indicating the need for pursuing the immunization programs. We also found high rates of vaccination coverage in the Western Brazilian Amazon region. PMID- 15361992 TI - Antiretroviral therapy during pregnancy and early neonatal life: consequences for HIV-exposed, uninfected children. AB - Women have emerged as the fastest growing human immunodeficiency virus (HIV) infected population worldwide, mainly because of the increasing occurrence of heterosexual transmission. Most infected women are of reproductive age and one of the greatest concerns for both women and their physicians is that more than 1,600 infants become infected with HIV each day. Almost all infections are a result of mother-to-child transmission of HIV. With the advent of combination antiretroviral therapies, transmission rates lower than 2% have been achieved in clinical studies. Antiretroviral compounds differ from most other new pharmaceutical agents in that they have become widely prescribed in pregnancy in the absence of proof of safety. We reviewed antiretroviral agents used in pregnant women infected with human immunodeficiency virus, mother-to-child transmission, and their consequences for infants. PMID- 15361993 TI - Evaluation of the rapid diagnostic test OptiMAL for diagnosis of malaria due to Plasmodium vivax. AB - OBJECTIVE: To determine the sensitivity and specificity of the rapid diagnostic test OptiMAL for diagnosis of Plasmodium vivax malaria. MATERIAL AND METHODS: We included all the patients who sought medical attention in the San Martin Pangoa Hospital, Junin, an area endemic for vivax malaria in Peru, between October and December 1998, who had fever during the previous 72 hours and who were older than 12 months. The gold standard for diagnosis was thick blood film microscopy. We determined the parasitemia rate for each of the positive slides. We calculated sensitivity, specificity, positive predictive value and negative predictive value of the test. RESULTS: We included 72 patients; 39 of them were positive for P. vivax by microscopic examination. The sensitivity of the Optimal test was 92.3%, the specificity 100%, the positive predictive value 100% and the negative predictive value 91.6%. The accuracy of the test was 95.8%. The sensitivity of the OptiMAL test progressively decreased when parasitemia was lower than 1,000 parasites/microliter. CONCLUSIONS: the OptiMAL test has a high sensitivity and specificity for diagnosis of P. vivax malaria. However, its sensitivity decreased when parasitemia levels were lower. It is a very simple technique, which makes it a good alternative for malaria diagnosis in remote places, although its elevated cost is still a problem. PMID- 15361994 TI - Aminotransferase changes and acute hepatitis in patients with dengue fever: analysis of 1,585 cases. AB - INTRODUCTION: Type 3 dengue virus caused an extensive epidemic in the state of Rio de Janeiro in summer 2002. In some of the patients, it was found in an atypical form with increased aminotransferase levels and acute hepatitis. MATERIAL AND METHODS: An analysis was made of 1,585 serologically confirmed dengue cases at the Dengue Reference Center in Campos dos Goytacazes, Rio de Janeiro state. The grade of hepatic aggression was established according to the alterations in the aminotransferase levels: grade A -- normal levels of aminotransferase; grade B -- elevated aminotransferase, with increased levels of at least one of the enzymes; grade C -- elevated aminotransferase, with the levels of at least one of the enzymes increased to more than three times the reference values; grade D -- acute hepatitis, with aminotransferase levels increased to at least 10 times their normal values. RESULTS: Among the 1,585 serologically confirmed dengue cases, 44.5% presented alterations in the aminotransferase levels (grade B), 16.9% presented grade C liver involvement and 3.8% of the patients had progressed to acute hepatitis (grade D). The average values for the rise in aspartate aminotransferase and alanine aminotransferase were 93.3 U/L and 86.0 U/L. The greatest alterations were observed among females (p<0.001), cases of dengue hemorrhagic fever (p<0.001), and cases with sequential infections (p=0.001). CONCLUSIONS: Liver damage with elevation of aminotransferases and reactive hepatitis was a common complication of dengue virus infection in these patients. PMID- 15361995 TI - Risk factors for Toxoplasma gondii infection in women of childbearing age. AB - OBJECTIVES: Determine the risk factors involved in toxoplasmosis transmission and determine whether pregnancy is a risk factor for toxoplasmosis infection. STUDY DESIGN: Cross-sectional study carried out on 2,242 women at childbearing age. An indirect immunofluorescence reaction was used to identify immunity to Toxoplasma gondii. Previous gestations were also analyzed as a possible risk factor. The results were analyzed by chi2 and OR tests, and by variance analysis. The sample was statistically balanced according to social-economic risk factors. RESULTS: Previously pregnant women were 1.74 times more frequently infected with toxoplasmosis, regardless of environmental conditions. Pregnant women living under unfavorable environmental conditions had an approximately two times increased risk of being infected for each risk factor (contact with host animals, presence of vehicles of oocyst transmission). Previous pregnancy was the risk factor that had the strongest influence on acquiring toxoplasmosis (variance analysis and statistical balancing). DISCUSSION: The prevalence of this zoonosis is high in Goiania-GO, Brazil (65.8%). Inadequate environmental sanitation was not significantly correlated with toxoplasmosis infection, except when associated with previous pregnancy, showing that the fundamental cause for infection is not environmental. CONCLUSION: The finding that pregnancy makes women more vulnerable to this protozoan, makes it important to implement prophylactic control of at risk pregnant women. PMID- 15361996 TI - Cerebral infarction related to cryptococcal meningitis in an HIV-infected patient: case report and literature review. AB - Neurological dysfunction as the first manifestation of AIDS has been found in 10 to 20% of symptomatic human immunodeficiency virus infections. However, stroke has rarely been reported in AIDS patients. The most common causes of cerebral infarction in AIDS are central nervous system infections: toxoplasmosis, cryptococcal meningitis and tuberculosis. Potential vascular mechanisms for cerebral infarction and transient neurological deficits among AIDS patients include deposition of antigen-antibody complexes with vasculitis and infarction, and a direct toxic effect of a viral antigen or infectious agent on vascular endothelium. The role of cryptococcal meningitis in vasculopathy is still not clear. We report a case of cerebral infarction in an HIV-infected patient, with cryptococcal meningitis as the first manifestation of AIDS. PMID- 15361997 TI - Acute liver failure complicating viral hepatitis A. AB - Hepatitis A is one of the most frequent infectious liver diseases affecting children worldwide. The disease is usually mild and self-limited, and complications are very rare. Nevertheless, hepatitis A can sometimes cause acute liver failure (ALF), a severe, life-threatening condition. Herein is reported a case of a child who presented ALF during a course of hepatitis A. The need for early identification of possible ALF cases among hepatitis A patients, and for effective ways of evaluating such a possibility, are discussed. We also emphasize the importance of prevention measures, especially vaccination. PMID- 15361998 TI - Mycobacterium tuberculosis bacteremia diagnosed in an HIV-negative patient in Brazil: a rare or an under-reported event? AB - A case of Mycobacterium tuberculosis bacteremia in an HIV negative immunodepressed patient was described using the BACTEC 460 TB system. This bacterium should be investigated in the blood of immunodepressed non-HIV infected patients with prolonged fever. PMID- 15361999 TI - Actinomycosis simulating malignant large bowel obstruction. AB - We present a case of a 58 year old white male who entered the hospital with abdominal pain and developed large bowel obstruction, simulating malignant disease. Anatomopathological examination showed abdominal actinomycosis, a rare presentation of this disease. PMID- 15362001 TI - [Mental disorders in the parents of bipolar patients]. AB - OBJECTIVE: It is well known that bipolar disorder has familial transmission. Studies indicate that first-degree relatives of bipolar patients also have higher incidences of other mental disorders than the general population. The aim of this study is to determine the prevalence of mental disorders in the parents of bipolar patients. METHOD: Parents of 35 probands who were treated for their bipolar disorders and 35 age and gender matched healthy subjects' parents were assessed by means of the Structured Clinical Interview for DSM IV (SCID-I), and a questionnaire for the parents. RESULTS: Past and current mental disorders were present in 27.1% of the parents of bipolar patients but in 14.2% of the parents of normal controls; the difference between the groups is statistically significant. The most prevalent mental disorder in both groups is major depressive disorder. The parents of bipolar patients were more likely to have a family history of mental disorder (44.2%) than the control group (12.8%). Parents who had a current or past mental disorder were more likely to have a family history of mental disease. Offspring of in-bred families had more bipolar siblings. Bipolar children of parents who had a family history of mental disorder tended to have an earlier age of onset. CONCLUSION: The findings of this study indicate that there is an increased prevalence of bipolar disorders and other mental disorders in the parents of bipolar patients when compared to the parents of healthy controls. PMID- 15362002 TI - [Is there a relationship between mood disorders and affective temperaments?]. AB - OBJECTIVE: The aim of this study is to investigate whether depressive (DT), hyper thymic (HT), cyclothymic (CT), irritable and anxious temperaments as identified by Turkish version of Temperament Evaluation of Memphis, Pisa, Paris and San Diego Auto questionnaire (TEMPS-A) are characteristic to depressive or bipolar disorder (BD) and to compare underlying affective temperamental differences. METHOD: 68 patients with recurrent major depression (MD-R), 50 patients with single episode major depressive disorder (MD-S), 84 euthymic bipolar patients and 100 healthy controls were included in the study and evaluated with TEMPS-A. RESULTS: There was a gender difference between patient groups. The females were more dominant in the MD-R group. The mean age of onset of illness of bipolar patients was lower than the depressive patients and the mean duration of illness of bipolar patients was longer than those of the MD-R patients. The prevalence of the rate of any dominant affective temperament was significantly higher in the MD R group than other study groups. The prevalence rate and scores of DT were significantly higher both in MD-R and MD-S groups (25.0% and 12.0% respectively) than the BD group (2.4%) and control subjects (3.0%). The prevalence rate and scores of CT were higher among the patients groups than the controls. None of the subjects except BD patients had HT. CONCLUSION: With the limitations of the study, it is thus reasonable to speculate that affective temperament, to a degree, determine the nature and the existence of the mood disorders. PMID- 15362003 TI - [Alexithymia and anger in women with fibromyalgia syndrome]. AB - OBJECTIVE: Fibromyalgia syndrome is characterized by both somatic and psychic symptoms and it is suggested that psychic factors contribute to the clinical presentation of this syndrome. This study was planned to have a better understanding of fibromyalgia through elaborating the role of alexithymia and anger in the pathogenesis of this illness. METHOD: The study was carried out in the Physical Therapy and Rehabilitation outpatient clinic with 101 women with fibromyalgia syndrome, 30 women with rheumatoid arthritis and 59 healthy women with no current or past medical history. The subjects were evaluated by the Visual Analog Scale, Toronto Alexithymia Scale-20 items, Spielberger State-Trait Anger Inventory, Beck Depression Scale, Fibromyalgia Impact Questionnaire and a sociodemographic data form. RESULTS: All these groups were similar to each other in means of age, years of education, marital and economical status. In the fibromyalgia syndrome group, the scores of anxiety and anger-in were calculated significantly higher than other groups. The depression and alexithymia scores were found higher than healthy group. DISCUSSION: These findings suggest that fibromyalgia patients suffer more anxiety and anger toward oneself, which is anger-in, than rheumatoid arthritis patients. Though the patient groups were more alexithymic than the healthy group, alexitimia scores of the two patient groups were not different. This situation suggest that anger-in, which is suppressed and unexpressed anger style is a part of the fibromyalgia syndrome together as well as high anxiety. PMID- 15362004 TI - [Depression-anxiety and disability in the premenopausal and postmenopausal period]. AB - OBJECTIVE: The aim of the study is to compare levels of depression-anxiety and disability in premenopausal and postmenopausal women. METHOD: The study was carried out with women between 45-55 years of age, living in the service area of a Mother-Child Health and Family Planning Center. Women who did not have a menstruation at least for one year were accepted as postmenopausal. The premenopausal group was consisted of women having menstruation. Hospital Anxiety and Depression Scale (HAD) and Brief Disability Questionnaire (BDQ) were applied to the women. RESULTS: Four hundred and ninety four women were recruited to the study. There were 214 women in the premenopausal period and 280 women in the postmenopausal period. In premenopausal women the level of education was higher and smoking was more prevalent; whereas in postmenopausal women the prevalence rates of being housewife and having continuous drug therapy due to chronic physical diseases were higher. In postmenopausal women, levels of depression and disability were significantly higher. In this group, the predictive factors for having a high HAD score were disability, education having a previous psychiatric disorder having drug therapy due to chronic physical diseases, diabetes mellitus, low level, menopause before the age of 40, not having any counselling for menopause, and not having routine laboratory tests. Having a previous psychiatric disorder and not having any counselling were also predicting disability. None of the variables predicted a high HAD score psychiatric disorder and not having any counselling were also predicting disability and disability in the premenopausal women. CONCLUSION: The postmenopausal period seems to predispose women to depressive symptoms and disability compared to the premenopausal period. PMID- 15362005 TI - [Reliability and validity of Turkish form of Endicott Work Productivity Scale]. AB - OBJECTIVE: The Endicott Work Productivity Scale (EWPS) is a brief self-report questionnaire designed to enable investigators to obtain a sensitive measure of work productivity. The purpose of the study was to investigate reliability and validity of the Turkish form of EWPS. METHOD: The study was performed with two separate groups; a patient group (n=74) and a community sample (n=107). Through the adaptation process the original form was translated into Turkish, and translated back into English. Statistical tests about construct validity, the concurrent validity, the test re-test reliability and internal consistency were performed. RESULTS: The internal consistency coefficient (Cronbach's alpha) was >.90 for both groups. The correlations of the individual items with the EWPS total scores ranged between.32 and.75. EWPS and HAM-D scores of the patient group was positively correlated (r =.52 and r =.62). The CGI scores correlation was.64 and SF-36 social functioning subscale was correlated negatively (r= -.43). The correlations of the test-retest scores of EWPS were.76. There was a significant mean difference between the total scores of patient and the community groups. CONCLUSION: The Turkish form of EWPS was found to be valid and reliable. The findings concerning the reliability and the validity of the EWPS indicate that this instrument can be used in the studies that will be conducted in Turkey. PMID- 15362006 TI - [Panic-agoraphobic spectrum]. AB - Current diagnostic classification systems have provided reliable, objective and valid diagnoses of mental disorders. They present both categorical and dichotomous approaches to mental disorders. However, rigid usage of diagnostic criteria may cause high comorbidity rates. Moreover, these classification systems fail to identify subthreshold conditions, atypical signs and symptoms, and personality traits associated with the core symptoms of a given mental disorder. These subclinical manifestations are clinically meaningful in terms of disability and effects on quality of life. For these reasons, many researchers have begun to develop dimensional or spectral approaches for a number of mental disorders. Studies on panic disorder with or without agoraphobia have revealed a substantial clinical heterogeneity. The panic-agoraphobic spectrum concept was defined to overcome these difficulties. This concept includes DSM-IV and ICD-10 diagnostic criteria together with atypical and subthreshold symptoms of panic disorder, and constitutes a broader band of panic and agoraphobia symptoms. The panic agoraphobic spectrum incorporates eight domains of clinical features: 1) separation sensitivity, 2) panic-like symptoms, 3) stress sensitivity, 4) medication and substance sensitivity, 5) anxious expectation, 6) agoraphobia, 7) illness phobia and hypochondriasis, 8) reassurance orientation. In this article, we reviewed the spectrum conceptualization of panic disorder and agoraphobia, particularly focusing on the domains of the panic-agoraphobic spectrum and their assessment. PMID- 15362007 TI - [Anaesthesia in electroconvulsive therapy]. AB - Electroconvulsive therapy (ECT) is a safe and efficient procedure performed for the treatment of drug resistant depression and other psychiatric disorders. Nowadays, its administration under general anaesthesia is a worldwide process. Anaesthetic management generally involves a short acting barbiturate such as thiopental in induction followed by a muscle relaxant. Succinylcholine is the drug of choice for muscle relaxation. This approach prevents patients from suffering hazardous orthopaedic injuries due to confusion. The effectiveness of ECT depends on an adequate seizure, and so the anaesthetist should be aware of the factors that influence the duration of seizures as well as concomitant diseases and potential adverse antidepressive drug reactions. An acute haemodynamic response due to sympathetic discharge in the course of the seizure provokes abrupt cardiovascular and cerebrovascular changes such as bradycardia, tachycardia, hypertension and raised intracranial pressure. The control of responses by b-blockers and similar agents is especially important in patients with cardiac or intracerebral problems. ECT is applicable in nearly all age groups and even in pregnant subjects. The aim of this article is to review the aspects of anaesthetic management of safe and effective ECT. PMID- 15362008 TI - [Psychiatric disorders and disability among leprosy patients; a review]. AB - As a gross estimate, leprosy currently affects 11-16 million patients worldwide There are currently 3600 registered patients in Turkey. The social stigma connected to leprosy makes this disease completely different from others. Even nowadays people affected by leprosy have to leave their village or are socially isolated. The physical deformity ratio is approximately 25% in other countries whereas it is more than half in Turkey. The prevalence of mental disorders among leprosy patients is higher than that among the general population. Depression is the most common psychiatric disorder among leprosy patients. Another important finding is that the long duration of the illness and physical handicaps raise the risk of psychiatric disorders. Nevertheless, the results of two studies conducted in Turkey on this subject contradict the results of international studies. Leprosy patients experience functional disabilities that limit their lives and ability to establish relation ships with others both in social and occupational fields. The physical disability rate is high (75%) in Turkey. A review of the literature revealed several papers on the psychosocial aspects of illness but few references to the degree or pattern of psychiatric disorders among leprosy patients. The main purpose of this paper is to review psychiatric disorders and disabilities in leprosy patients and to obtain concrete results. PMID- 15362009 TI - Fall risk factors assessment tool: enhancing effectiveness in falls screening. AB - A quasi-experimental study was conducted to explore the effectiveness of fall prevention among hospital patients based on the modified fall risk factors assessment tool. We investigated the frequency of falls among hospital patients at a medical center in Taiwan. The experimental group of falls victims was selected from patients (n = 39) hospitalized in 2002 after falls. The control group of patients falls was selected by means of a retrospective incident report review which identified patients (n = 43) hospitalized one year earlier. The results showed that there was no significant difference in the incidence of falls between the two groups. Nevertheless, there were significant differences in age, indications of falls, use of sedatives, walking ability and evaluated grade of fall risk factors. In addition, the average level of satisfaction under recently modified fall risk factors evaluation guideline was 2.68 points (upper limit = 4 points) based upon investigation derived from nursing staff ' s opinions. Moreover, nursing staff from GYN/OBS and orthopedics departments acknowledged the enhanced effectiveness of these new guidelines. The screening rate for high-risk orthopedic patients was increased from 20.7 % to 41.9 %. Furthermore, the screening rate among the experimental group (74.4 % ) was also higher than that among the control group (60.5 % ) ( p <.01). In line with our effective tool to screen high-risk patients, we also added the concept of continuous quality improvement in nursing care to implement a fall prevention program to reduce unnecessary injury. This strategy may assist nursing personnel in providing immediate and individualized care as well as health education for high-risk patients. It may also cause the incidence of patient falls in hospitals to continue to decline. PMID- 15362010 TI - Exercise behavior and related factors in career women - the case of a bank in Taipei City. AB - With the trend of premature aging of physiological functions on the rise and a variety of chronic diseases continuing to spread, health promotion has become the top concern among public health experts. Regular exercise plays a pivotal role in both health promotion and disease prevention. This study aims to investigate the exercise behavior of career women and related factors. The samples were drawn from the female employees of a bank in Taipei, totaling 361 persons, all aged between 20 and 56. The result shows that only 8.6 % of the respondents exercise regularly and that among the reasons for not doing any exercise, " Don ' t have time for it " tops the list. Self-efficacy in exercise is found to be the common factor for predicting both exercise regularity and total exercise amount. Exercise intervention programs thus must be developed on the basis of female self efficacy with a " family-oriented " activity design. It is therefore suggested that employers promote exercise and encourage exercise behaviors to help enhance employee self-efficacy as well as employee health. PMID- 15362011 TI - A comparative study of expectant parents ' childbirth expectations. AB - The purpose of this study was to understand childbirth expectations and differences in childbirth expectations among expectant parents. For convenience sampling, 200 couples willing to participate in this study were chosen from two hospitals in central Taiwan. Inclusion criteria were at least 36 weeks of gestation, aged 18 and above, no prenatal complications, and willing to consent to participate in this study. Instruments used to collect data included basic demographic data and the Childbirth Expectations Questionnaire. Findings of the study revealed that (1) five factors were identified by expectant parents regarding childbirth expectations including the caregiving environment, expectation of labor pain, spousal support, control and participation, and medical and nursing support; (2) no general differences were identified in the childbirth expectations between expectant fathers and expectant mothers; and (3) expectant fathers with a higher socioeconomic status and who had received prenatal (childbirth) education had higher childbirth expectations, whereas mothers displayed no differences in demographic characteristics. The study results may help clinical healthcare providers better understand differences in expectations during labor and birth and childbirth expectations by expectant parents in order to improve the medical and nursing system and promote positive childbirth experiences and satisfaction for expectant parents. PMID- 15362012 TI - An investigation on the nursing competence of southern Taiwan nurses who have passed N3 case report accreditation. AB - The purpose of this study was to examine factors affecting the development of nursing competence in the writing of N3 (clinical nurse level III) case reports. A cross-sectional survey was conducted in 13 hospitals in southern Taiwan from July to October, 2002. The subjects included 239 nurses who had passed N3 case report accreditation. The results show that (1) nurses rated their current competence at a fairly good level ( 5.57 +/- 0.74). There was no significant difference between nurses ' subjective self-evaluation and supervisors ' objective evaluation (5.57 +/- 0.74 vs 5.55 +/- 0.89; t = 0.162, p = 0.871). (2) The nurses ' self-rating, supervisors ', and colleagues ' evaluation all indicated moderate progress in nursing competence after writing N3 case reports. The progress scores rated by nurses themselves, supervisors and colleagues were not significantly different (4.89 +/- 1.35 vs. 4.84 +/- 1.57 vs. 4.84 + 1.26; F = 0.096, p =.909). This finding reveals that writing N3 case reports has a positive effect on nursing competence. (3) The results also indicate that ' Attitude toward N3 case report ', 'Writing difficulty ', ' Preparation for writing case report ' were the three predicting factors of progress in nursing competence, which accounted for 26.2 % of the total variance. Therefore we recommend that N3 advancement education should include guidance in alleviating writing difficulty and emphasis on implementing various types of preparation for writing case reports, as effective ways to improve nursing competence. PMID- 15362013 TI - Important computer competencies for the nursing profession. AB - Nursing requires computer competencies. This study aimed at identifying those competencies required for the nursing profession in Taiwan. The Delphi technique was deployed in this study. In the Delphi questionnaires, computer competencies were sorted into seven domains: concepts of hardware, software, and networks; principles of computer applications; skills of computer usage; program design; limitations of the computer; personal and social issues; attitudes toward the computer. In three Delphi questionnaires, nursing informatics experts gave us their opinions on the importance of each computer competency for the nursing profession. The experts also designated when the competency should be cultivated. This study provides a comprehensive list for nursing professionals to check on their computer competence. The results of this study should also serve as good references for teachers and schools in designing related curriculums. PMID- 15362014 TI - The development of intelligent, triage-based, mass-gathering emergency medical service PDA support systems. AB - The support systems for the Emergency Medical Services (EMS) at mass gatherings, such as the local marathon or large international baseball games, are underdeveloped. The purposes of this study were to extend well-developed, triage based, EMS Personal Digital Assistant (PDA) support systems to cover pre-hospital emergency medical services and onsite evaluation forms for the mass gatherings, and to evaluate users ' perceived ease of use and usefulness of the systems in terms of Davis ' Technology Acceptance Model (TAM). The systems were developed based on an established intelligent triage PDA support system and two other forms the general EMS form from the Taipei EMT and the customer-made Mass Gathering Medical form used by a medical center. Twenty-three nurses and six physicians in the medical center, who had served at mass gatherings, were invited to examine the new systems and answer the TAM questionnaire. The PDA systems were composed of 450 information items within 42 screens in 6 categories. The results supported the potential for using triage-based PDA systems at mass gatherings. Overall, most of the subjects agreed that the systems were easy to use and useful for mass gatherings, and they were willing to accept the systems. PMID- 15362015 TI - The comparative effectiveness among institutionalized and non-institutionalized elderly people in Taiwan of reminiscence therapy as a psychological measure. AB - This study examined the comparative effects of reminiscence on self-esteem, self health perception, depressive symptoms, and mood status of elderly people residing in long-term care facilities and at home. A quasi-experimental design was conducted, using pre-intervention and post-intervention tests and purposive sampling. Rosenberg ' s Self-Esteem Scale (RSE), Health Perception Scale (HPS), Geriatric Depression Scale Short Form (GDS-SF), and Apparent Emotion Rating Scale (AER) were used as study instruments. Each subject was administered pre- and post experimental tests at a four month interval and all subjects underwent weekly individual reminiscence intervention. Forty-eight subjects completed the study, with 25 institutionalized elderly people and 23 non-institutionalized home-based elderly people. Independent t-tests and paired t-tests were conducted to measure the differences in variable means between and within groups. A significant difference was found between groups in mood status post-test (t = 5.96, p <.001) and significant differences were noted in self-health perception, depressive symptoms, and mood status (t = -2.56, 2.83, -3.02; p =.018,.009,.007) between the pre- and post-intervention tests in the institutionalized group. These results suggest that reminiscence therapy is especially appropriate for older people who reside in care facilities. Implementing strategies that enrich the lives of elderly people residing in long-term cares is crucial, and reminiscence offers a method for promoting healthy aging. PMID- 15362016 TI - The effects of continuing education in restraint reduction on novice nurses in intensive care units. AB - A decrease in the use of physical restraints in Intensive Care Units (ICUs) is an important indicator of quality of nursing care. This quasi-experimental study examined the effect on nurses of a session of continuing education aimed at reducing the use of restraints. At a medical center in southern Taiwan, 37 novice nurses were surveyed about their knowledge, perception, attitude and clinical practice of restraint use in 11 ICUs. Two instructors then taught a four-hour continuing education class on patient restraint standards, principles for reducing physical restraint use, and alternatives to restraining. Data were analyzed by paired t-test and the results of identical structured questionnaires which participants received before and after the lecture, showed that, afterwards, knowledge (t = -6.04, p <.01), perception (t = 4.76, p <.01), and attitude (t = 3.93, p <.01) toward restraint use had significantly improved. The continuing education improved the nurses ' knowledge and attitude toward restraint use and may therefore enhance the quality of care provided to ICU patients. PMID- 15362017 TI - Association of the DTNBP1 locus with schizophrenia in a U.S. population. AB - Linkage and association studies have recently implicated dystrobrevin-binding protein 1 (DTNBP1) in the etiology of schizophrenia. We analyzed seven previously tested DTNBP1 single-nucleotide polymorphisms (SNPs) in a cohort of 524 individuals with schizophrenia or schizoaffective disorder and 573 control subjects. The minor alleles of three SNPs (P1578, P1763, and P1765) were positively associated with the diagnosis of schizophrenia or schizoaffective disorder in the white subset of the study cohort (258 cases, 467 controls), with P1578 showing the most significant association (odds ratio 1.76, P =.0026). The same three SNPs were also associated in a smaller Hispanic subset (51 cases, 32 controls). No association was observed in the African American subset (215 cases, 74 controls). A stratified analysis of the white and Hispanic subsets showed association with the minor alleles of four SNPs (P1578, P1763, P1320, and P1765). Again, the most significant association was observed for P1578 (P =.0006). Haplotype analysis supported these findings, with a single risk haplotype significantly overrepresented in the white sample (P =.005). Our study provides further evidence for a role of the DTNBP1 gene in the genetic etiology of schizophrenia. PMID- 15362023 TI - Pharmaceutical influence on clinical science and practice: a double-edged sword. PMID- 15362024 TI - Image of the month. Gallstone ileus. PMID- 15362025 TI - Confocal laser endoscopy for diagnosing intraepithelial neoplasias and colorectal cancer in vivo. AB - BACKGROUND & AIMS: A confocal laser endoscopy system has recently been developed that may allow subsurface imaging of living cells in colonic tissue in vivo. The aim of the present study was to assess its potential for prediction of histology during screening colonoscopy for colorectal cancer. METHODS: Twenty-seven patients underwent colonoscopy with the confocal endoscope using acriflavine hydrochloride or fluorescein sodium with blue laser illumination. Furthermore, 42 patients underwent colonoscopy with this system using fluorescein sodium. Standardized locations and circumscript lesions were examined by confocal imaging before taking biopsy specimens. Confocal images were graded according to cellular and vascular changes and correlated with conventional histology in a prospective and blinded fashion. RESULTS: Acriflavine hydrochloride and fluorescein sodium both yielded high-quality images. Whereas acriflavine hydrochloride strongly labeled the superficial epithelial cells, fluorescein sodium offered deeper imaging into the lamina propria. Fluorescein sodium was thus used for the prospective component of the study in which 13,020 confocal images from 390 different locations were compared with histologic data from 1038 biopsy specimens. Subsurface analysis during confocal laser endoscopy allowed detailed analysis of cellular structures. The presence of neoplastic changes could be predicted with high accuracy (sensitivity, 97.4%; specificity, 99.4%; accuracy, 99.2%). CONCLUSIONS: Confocal laser endoscopy is a novel diagnostic tool to analyze living cells during colonoscopy, thereby enabling virtual histology of neoplastic changes with high accuracy. These newly discovered diagnostic possibilities may be of crucial importance in clinical practice and lead to an optimized rapid diagnosis of neoplastic changes during ongoing colonoscopy. PMID- 15362026 TI - The incidence of colorectal cancer following a negative screening sigmoidoscopy: implications for screening interval. AB - BACKGROUND & AIMS: Current guidelines recommend a 5-year interval for colorectal cancer (CRC) screening by sigmoidoscopy. However, the optimal screening interval is uncertain. We estimated the annual incidence of distal and proximal CRC in the first 5 years following a negative sigmoidoscopy examination to gauge the potential benefit of rescreening in <5 years. METHODS: A cohort of 72,483 participants in the Colon Cancer Prevention program of Kaiser Permanente of Northern California (KP) was defined using computerized databases. Men and women aged 50 years and older who had a negative screening flexible sigmoidoscopy examination between 1994 and 1996 and were considered not to be at high risk for developing CRC were included. Subjects were censored at the time of diagnosis (for cases), death, termination of KP membership, or subsequent colon examination. RESULTS: Thirty cases of distal and 80 cases of proximal CRC occurred. Age-adjusted incidence rates of distal CRC ranged from a low of 2.8 per 100,000 person-years in the first year of follow-up to a high of 13.0 per 100,000 in the fourth year (rate difference, 10.2; 95% confidence interval, 1.1-19.3). However, for the entire follow-up period, incidence of distal CRC remained much lower than age-adjusted rates of 70.6 in the general population (Surveillance, Epidemiology, and End Results registry). The incidence of proximal CRC was also decreased modestly over population rates of disease. CONCLUSIONS: Screening by sigmoidoscopy more frequently than every 5 years would likely lead, at best, to only modest improvements as compared with a 5-year screening interval. PMID- 15362027 TI - Postoperative maintenance of Crohn's disease remission with 6-mercaptopurine, mesalamine, or placebo: a 2-year trial. AB - BACKGROUND & AIMS: No therapy has been shown to reliably prevent the evolution of postoperative recurrence of Crohn's disease. The aim of the current trial was to compare 6-mercaptopurine (6-MP) and mesalamine with placebo for the prevention of clinical, endoscopic, and radiographic recurrence of Crohn's disease after resection and ileocolic anastomosis. METHODS: Five centers randomized 131 patients to receive 6-MP (50 mg), mesalamine (3 g), or placebo daily in a double blind, double-dummy trial. Patients had clinical assessments at 7 weeks and then every 3 months; colonoscopy at 6, 12, and 24 months; and small bowel series at 12 and 24 months. End points were clinical, endoscopic, and radiographic recurrence rates at 24 months. RESULTS: Clinical recurrence rates (intent to treat) by life table analysis at 24 months were 50% (95% confidence interval [CI], 34%-68%), 58% (95% CI, 41%-75%), and 77% (95% CI, 61%-91%) in patients receiving 6-MP, mesalamine, and placebo, respectively. Endoscopic recurrence rates were 43% (95% CI, 28%-63%), 63% (95% CI, 47%-79%), and 64% (95% CI, 46%-81%), and radiographic recurrence rates were 33% (95% CI, 19%-54%), 46% (95% CI, 29%-66%), and 49% (95% CI, 30%-72%), respectively. 6-MP was more effective than placebo ( P < 0.05) at preventing clinical and endoscopic recurrence over 2 years. Patient withdrawals resulted in 69% of the study population evaluable for the clinical recurrence end point. CONCLUSIONS: 6-MP, 50 mg daily, was more effective than placebo at preventing postoperative recurrence of Crohn's disease and should be considered as a maintenance therapy after ileocolic resection. PMID- 15362028 TI - Azathioprine and mesalamine for prevention of relapse after conservative surgery for Crohn's disease. AB - BACKGROUND & AIMS: Because the reoperation rate for Crohn's disease is high after resective surgery, use of conservative surgery has increased. Mesalamine was investigated for the prevention of postoperative relapse, with disappointing results. The role of azathioprine in the postoperative setting is unknown. We aimed to compare the efficacy and safety of azathioprine and mesalamine in the prevention of clinical and surgical relapse in patients who have undergone conservative surgery for Crohn's disease. METHODS: In a prospective, open-label, randomized study, 142 patients received azathioprine (2 mg. kg -1. day -1 ) or mesalamine (3 g/day) for 24 months. Clinical relapse was defined as the presence of symptoms with a Crohn's Disease Activity Index score >200 and surgical relapse as the presence of symptoms refractory to medical treatment or complications requiring surgery. RESULTS: After 24 months, the risk of clinical relapse was comparable in the azathioprine and mesalamine groups, both on intention-to-treat (odds ratio [OR], 2.04; 95% confidence interval [CI], 0.89-4.67) and per-protocol analyses (OR, 1.79; 95% CI, 0.80-3.97). No difference was observed with respect to surgical relapse at 24 months between the 2 groups. In a subgroup analysis, azathioprine was more effective than mesalamine in preventing clinical relapse in patients with previous intestinal resections (OR, 4.83; 95% CI, 1.47-15.8). More patients receiving azathioprine withdrew from treatment due to adverse events than those receiving mesalamine (22% vs. 8%; P = 0.04). CONCLUSIONS: While no difference was observed in the efficacy of azathioprine and mesalamine in preventing clinical and surgical relapses after conservative surgery, azathioprine is more effective in those patients who have undergone previous intestinal resection. PMID- 15362029 TI - Shiner of the Shiner mucosal biopsy tube. PMID- 15362030 TI - A circulating ligand for galectin-3 is a haptoglobin-related glycoprotein elevated in individuals with colon cancer. AB - BACKGROUND & AIMS: Galectin-3 is a beta-galactoside-binding protein implicated in tumor progression and metastasis of colorectal cancers. To determine whether circulating galectin-3 ligands are related to the presence of colon cancer, we sought to identify and quantify ligands in serum that bind to galectin-3. METHODS: Sera from patients with colon cancer, adenomas, and normal individuals were desialylated, reduced, and separated by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) and blots probed with biotinylated galectin-3. RESULTS: In colon cancer sera, the major galectin-3 ligand was a 40-kilodalton band distinct from mucin, carcinoembryonic antigen, and Mac-2 binding protein. Serum 40-kilodalton ligand was 10- to 30-fold higher in patients with colon cancer than in healthy subjects. Ligand was purified by gel filtration, affinity precipitation on galectin-3/agarose, and SDS-PAGE. When tryptic peptides were analyzed by matrix-assisted laser-desorption ionization mass spectrometry and protein database searching, the 40-kilodalton ligand was identified as haptoglobin beta subunit. In confirmation of this finding, depletion of haptoglobin by immunoprecipitation also eliminated the 40-kilodalton ligand. Colon cancer sera had only a modest increase in total haptoglobin as compared with healthy subjects, suggesting that the structure rather than the amount of haptoglobin is altered in patients with colon cancer. Immunohistochemical staining confirmed the absence of haptoglobin in normal colon and the ectopic expression of haptoglobin in colon cancers and adenomatous polyps. CONCLUSIONS: A major circulating ligand for galectin-3, which is elevated in the sera of patients with colon cancer, is a cancer-associated glycoform of haptoglobin. PMID- 15362031 TI - The impact of competing risks on the observed rate of chronic hepatitis C progression. AB - BACKGROUND & AIMS: In previous studies about the natural history of chronic hepatitis C (CHC), age at the time of infection correlated with the rate at which hepatic fibrosis progresses. The presence of a competing risk, namely higher mortality from natural causes, may contribute to this observation. A simulation experiment was conducted to measure the magnitude of the effect of competing risks on the observed rate of fibrosis progression of CHC. METHODS: A computer based probabilistic model was created in which fibrosis of CHC progressed from stage 0 to 4 (cirrhosis) in 20-year-old and 50-year-old male and female cohorts. The rate of fibrosis progression was randomly assigned to each simulated individual from a distribution common to all age- and sex-specific cohorts. The cohorts also experienced mortality from natural causes according to the 2000 census data. RESULTS: The observed median time to reach cirrhosis for the 50-year old cohorts was 20.4 +/- 0.2 years compared with 29.7 +/- 0.2 for the 20-year-old cohorts ( P < 0.01). The median time to reach cirrhosis in men was 24.2 +/- 0.6 years compared with 25.9 +/- 0.6 in women ( P = 0.01). Overall, the observed rate of progression was slowest among young women. Similarly, accelerating mortality from natural causes, simulating the impact of comorbid conditions that shorten survival, reduced the observed time to reach cirrhosis. CONCLUSIONS: Even if the underlying rate of fibrosis progression in CHC was held constant, the time to reach cirrhosis will be observed to be substantially shorter in subjects with a higher competing mortality. PMID- 15362032 TI - Natural history of chronic HBV carriers in northern Italy: morbidity and mortality after 30 years. AB - BACKGROUND & AIMS: Increased morbidity and mortality from liver disease have been reported in chronic hepatitis B surface antigen (HBsAg) carriers, but data on survival are equivocal. To assess the impact of hepatitis B virus (HBV) infection on survival and liver-related complications, we re-evaluated, after a mean follow up of 30 years, a cohort of 296 blood donors excluded from donation 30 years ago when HBsAg screening became mandatory. METHODS: Clinical and ultrasound examination and biochemical and virologic tests were performed. The cause of death was recorded and survival was compared with a control population of 157 HBV negative blood donors selected at baseline. RESULTS: Thirty-two (10.8%) cases and 14 controls (8.9%) ( P = 0.625) had died; 3 of 32 (9.3%) and 1 of 14 (7.1%) deaths were liver-related. Hepatocellular carcinoma (HCC) caused death in 2 of 296 and 1 of 157 subjects (0.6% in each group). Alcohol-induced cirrhosis occured in the remaining subject. By Cox regression analysis, survival was independently predicted by older age, abnormal gamma-glutamyl transpeptidase (GGT) levels, and presence of medical comorbidities at baseline. Unequivocal liver disease was found in 4 carriers only. No disease decompensation occurred during follow-up. Fifty-nine (32.2%) carriers cleared HBsAg (yearly incidence, 1.0%). Full-length serum HBV DNA was present in 32.2% of persistently HBsAg-positive individuals (average titer always <10 5 copies/mL). CONCLUSIONS: Over a 30-year period, chronic HBV carrier blood donors from Northern Italy did not develop clinically significant liver disease, hepatocellular cancer, or other liver-related morbidity or mortality at a higher rate than uninfected controls. The presence of medical comorbidities, older age at diagnosis, and abnormal GGT levels were independent predictors of death among chronic HBV carriers. PMID- 15362033 TI - Analysis of hepatitis C virus quasispecies transmission and evolution in patients infected through blood transfusion. AB - BACKGROUND & AIMS: Studies on hepatitis C virus (HCV) quasispecies dynamics in the natural course of infection are rare owing to difficulties in obtaining samples from the early phase of infection. METHODS: We studied 15 patients from the Transfusion-Transmitted Viruses Study who seroconverted to anti-HCV after receiving infected blood. Follow-up serum samples were collected every 2-3 weeks for 6 months, at 10 months, and at 11-16 years. Viral quasispecies in the second envelope hypervariable region 1 (E2/HVR1) and 5' untranslated region (5'UTR) were analyzed with single-strand conformation polymorphism (SSCP) and heteroduplex mobility assay (HMA). RESULTS: Seven patients cleared infection within 7-24 weeks (mean, 14.0 wk) and 3 patients eventually became anti-HCV negative. In 6 patients with resolving hepatitis the SSCP band pattern remained stable, whereas in one patient minor changes appeared before clearance. In contrast, in all 8 patients progressing to chronicity, major changes in the E2/HVR1 quasispecies developed at 8-22 weeks (mean, 13.1 wk). Shannon entropy and medium mobility shift values derived from HMA gels remained stable in patients with resolving hepatitis but changed in those who developed chronic infection. Only 2 patients showed minor changes in 5'UTR. A decrease in E2/HVR1 complexity at the time of transmission (bottleneck) was found in 5 patients altogether. CONCLUSIONS: Changes in E2/HVR1 quasispecies 8-22 weeks after infection, likely caused by mounting immune pressure, were predictive of ensuing chronic infection, whereas stability was associated with resolution. Our study also showed that composition of HCV quasispecies may be preserved during transmission from host to host. PMID- 15362034 TI - Activation of PPAR gamma and delta by conjugated linoleic acid mediates protection from experimental inflammatory bowel disease. AB - BACKGROUND & AIMS: The molecular targets for the protective actions of conjugated linoleic acid (CLA) on experimental inflammatory bowel disease (IBD) are unknown. We used a loss-of-function approach to investigate whether CLA ameliorated colitis through a peroxisome proliferator-activated receptor gamma (PPAR gamma) dependent mechanism. METHODS: The expression of PPAR gamma, delta, and their target genes in the colon of mice fed control or CLA-supplemented diets was assayed after a 7-day dextran sodium sulfate (DSS) challenge by quantitative real time polymerase chain reaction (PCR). Additionally, nuclear factor-kappa B (NF kappaB) p65 activation was quantified in the colon. To determine the involvement of PPAR gamma in the mechanism of action of CLA directly, specific deletions of PPAR gamma in the colon were performed in mice by using the Cre-lox recombination system. Colonic PPAR gamma null mice and wild-type littermates were fed either a CLA-supplemented or a control diet for 42 days and challenged with 2.5% DSS. The therapeutic efficacy of CLA also was examined by using the CD4 + CD45RB hi transfer colitis model. RESULTS: CLA induced PPAR gamma and delta, transcriptionally modulated PPAR gamma and delta-responsive gene clusters involved in lipid metabolism (uncoupling protein [UCP]1, UCP3, PPAR gamma coactivator 1alpha [PGC-1alpha], and CD36) and epithelial cell maturation (Gob-4 and Keratin 20). Additionally, CLA repressed tumor necrosis factor alpha (TNF alpha) expression and NF-kappaB activation while inducing the immunoregulatory cytokine transforming growth factor beta 1 (TGF-beta 1 ). Clinically, CLA ameliorated DSS- and CD4 + -induced colitis. Loss of the PPAR gamma gene in the colon abrogated the beneficial effects of CLA in DSS colitis. CONCLUSIONS: Our studies provide molecular evidence in vivo, suggesting that CLA ameliorates colitis through a PPAR gamma-dependent mechanism. PMID- 15362035 TI - Interleukin 10-deficient mice develop osteopenia, decreased bone formation, and mechanical fragility of long bones. AB - BACKGROUND & AIMS: Bone loss is a common complication of human inflammatory bowel disease (IBD), but its mechanisms are not understood completely. We investigated bone metabolism in interleukin-10-deficient ( IL-10-/- ) mice, an animal model with IBD features. METHODS: IL-10-/- male mice (8- and 12-weeks-old) and their age-matched wild-type counterparts (C57BL/6J) were studied. Bone mass of the femur was determined by ashing. Tibial cancellous and cortical bone mass and formation was measured by static and dynamic histomorphometry. Biomechanical strength of the femur was tested. Primary bone marrow stromal cell cultures were used to assess osteoblast generation. Serum levels of 25-OH vitamin D 3, insulin like growth factor-1 (IGF-1), parathyroid hormone, osteocalcin, and deoxy pyridinoline cross-links were measured. The presence of colitis was determined histologically, and by IL-12 and interferon-gamma (IFN-gamma) secretion from cultured colonic explants. RESULTS: Eight- and 12-week-old IL-10-/- mice developed osteopenia of both cancellous and cortical bone, evidenced by lower femoral ash weight, cancellous bone area and surface, trabecular number, and decreased cortical bone area and width. Osteopenia was associated with mechanical fragility, manifested by decreased stiffness and mechanical load at fracture, and was caused by suppressed bone formation, indicated by decreased cancellous double labeled surface, mineralizing surface, serum osteocalcin level, and mineralized nodule number in bone marrow stromal cell cultures. IL-10-/- mice with colitis had significantly less bone mass compared with IL-10-/- mice without colitis. CONCLUSIONS: IL-10-/- mice develop the hallmarks of osteoporosis, that is, reduced bone mass, increased mechanical fragility, and suppressed bone formation. The presence of colitis is an important contributor to osteoporosis in IL-10-/- mice. PMID- 15362036 TI - ClC-2 chloride secretion mediates prostaglandin-induced recovery of barrier function in ischemia-injured porcine ileum. AB - BACKGROUND & AIMS: Ischemia results in the breakdown of the intestinal barrier, predisposing patients to sepsis and multiple organ failure. Prostaglandins play a critical role in mediating recovery of barrier function in ischemia-injured intestine through a mechanism involving stimulation of Cl - secretion. In the present study, we investigated the contributory role of individual Cl - channels in the recovery of barrier function in ischemia-injured porcine ileum. METHODS: Ischemia-injured porcine ileal mucosa was mounted in Ussing chambers. Short circuit current (Isc) and transepithelial resistance (TER) were measured in response to prostaglandin E 2 (PGE 2 ) and pharmacologic inhibitors of epithelial Cl - channels. Immunoassays were used to assess the expression and localization of ion channels. RESULTS: Application of PGE 2 to ischemia-injured ileal mucosa stimulated increases in Isc, an indicator of Cl - secretion, that was followed by marked increases in TER, an indicator of barrier function recovery. In vitro studies revealed that although PGE 2 induced Cl - secretion via at least 3 distinct secretory pathways, recovery of barrier function was initiated by Cl - secretion via ClC-2 Cl - channels co-expressed with occludin and localized to tight junctions within restituting epithelium. Intravenous administration of furosemide to pigs subjected to 1 hour of ileal ischemia impaired recovery of barrier function, as evidenced by decreased TER and increased mucosal-to-serosal 3 H-mannitol flux after a 2-hour reperfusion/recovery period, confirming an important role for Cl - secretory pathways in vivo. CONCLUSIONS: ClC-2-mediated intestinal Cl - secretion restores TER in ischemia-injured intestine. These data may provide the basis for targeted pharmacologic therapy for diseases associated with impaired barrier function. PMID- 15362037 TI - Transmembrane tumor necrosis factor is a potent inducer of colitis even in the absence of its secreted form. AB - BACKGROUND & AIMS: Tumor necrosis factor (TNF) is cleaved proteolytically from a 26-kilodalton transmembrane precursor protein into secreted 17-kilodalton monomers. Transmembrane (tm) and secreted trimeric TNF are biologically active and may mediate distinct activities. We assessed the consequences of a complete inhibition of TNF processing on the course of colitis in recombination activating gene (RAG)2 -/- mice on transfer of CD4 CD45RB hi T cells. METHODS: TNF -/- mice, transgenic for a noncleavable mutant TNF gene, were used as donors of CD4 T cells, and, on a RAG2 -/- background, also as recipients. Kinetics of disease development were compared in the absence of TNF, in the absence of secreted TNF, and in the presence of secreted and tmTNF. The analysis at the end of the observation period included the histopathologic assessment of the intestine and the localization of TNF and interferon gamma (IFNgamma)-expressing cells. RESULTS: The complete prevention of TNF secretion in tmTNF transgenic RAG2 -/- mice neither prevented nor delayed disease induction by transferred transgenic for a noncleavable transmembrane mutant of mouse TNF (tmTNF tg) CD4 CD45RB hi T cells. tmTNF expression by transferred CD4 T cells, however, was not required for disease induction because severe colitis and weight loss also were observed in tmTNF RAG2 -/- recipients of TNF -/- CD4 CD45RB hi T cells. In the presence of tmTNF, the absence of secreted TNF did not affect frequency and distribution of TNF and interferon-gamma messenger RNA (mRNA)-expressing cells. CONCLUSIONS: These results indicate that specific inhibitors of TNF processing are not appropriate for modulating the pro-inflammatory and disease-inducing effects of TNF in chronic inflammatory disorders of the intestine. PMID- 15362038 TI - Lactobacillus paracasei normalizes muscle hypercontractility in a murine model of postinfective gut dysfunction. AB - BACKGROUND & AIMS: The effects of probiotics on gut dysfunction in postinfective irritable bowel syndrome are unknown. We tested whether probiotics influence persistent muscle hypercontractility in mice after recovery from infection with Trichinella spiralis and analyzed the underlying mechanisms. METHODS: Mice were gavaged with Lactobacillus paracasei, Lactobacillus johnsonii, Bifidobacterium longum, or Bifidobacterium lactis in spent culture medium from days 10 to 21 after infection. Additional mice received heat-inactivated Lactobacillus paracasei, Lactobacillus paracasei -free spent culture medium, or heat inactivated Lactobacillus paracasei -free spent culture medium. Lactobacilli enumeration, immunohistochemistry, and cytokine detection (enzyme-linked immunosorbent assay) were performed. Mice were also treated with Lactobacillus paracasei or Lactobacillus paracasei -free spent culture medium from days 18 to 28 after infection. Contractility was measured on days 21 and 28 after infection. RESULTS: Lactobacillus paracasei, but not Lactobacillus johnsonii, Bifidobacterium lactis, or Bifidobacterium longum, attenuated muscle hypercontractility. This was associated with a reduction in the Trichinella spiralis -associated T-helper 2 response and a reduction in transforming growth factor-beta1, cyclooxygenase-2, and prostaglandin E 2 levels in muscle. Attenuation of muscle hypercontractility by Lactobacillus paracasei -free spent culture medium was abolished after heat treatment. Improvement of muscle hypercontractility at day 28 after infection was also observed after the administration of Lactobacillus paracasei or Lactobacillus paracasei -free spent culture medium from day 18 after infection. CONCLUSIONS: Probiotics show strain dependent attenuation of muscle hypercontractility in an animal model of postinfective irritable bowel syndrome. This likely occurs via both a modulation of the immunologic response to infection and a direct effect of Lactobacillus paracasei or a heat-labile metabolite on postinfective muscle hypercontractility. Lactobacillus paracasei may be useful in the treatment of postinfective irritable bowel syndrome. PMID- 15362039 TI - Ursodeoxycholate/Sulindac combination treatment effectively prevents intestinal adenomas in a mouse model of polyposis. AB - BACKGROUND & AIMS: Preclinical studies in animal models, human epidemiological data, and clinical trials in patients with adenomatous polyposis have consistently indicated that sulindac and other nonsteroidal antiinflammatory drugs or cyclooxygenase inhibitors have the greatest potential efficacy among current candidates for colon tumor chemopreventive agents. However, at highly effective doses they all have some risk of toxicity, and their therapeutic profile might be improved by use at lower, more tolerable doses, in combination with a second agent acting via other mechanisms. METHODS: Sulindac was tested in combination with ursodeoxycholic acid (ursodiol), a naturally occurring 7-B epimer of the bile component chenodeoxycholic acid, for prevention of adenomas in the Min mouse model of adenomatous polyposis. RESULTS: Ursodeoxycholic acid caused a dose-dependent decrease in the number of intestinal tumors. Unlike sulindac and other nonsteroidal anti-inflammatory drugs, which are quite beneficial in the distal intestine but are somewhat less effective in the proximal small intestine (especially the clinically important periampullary duodenum), ursodeoxycholate had equal efficacy throughout the entire intestine, both proximal and distal. Combined treatment with low-dose sulindac was less toxic, with normal weight gain and fewer gastrointestinal ulcerations than high dose sulindac. Combined treatment with sulindac and ursodeoxycholate was more effective than either agent alone for the prevention of tumors throughout the entire intestine. CONCLUSIONS: These experiments provide the first evidence that ursodeoxycholic acid is effective for preventing adenomas in an animal model. Cyclooxygenase inhibition, when combined with this naturally occurring bile component, may become a promising approach for colon cancer prevention. PMID- 15362040 TI - Helicobacter pylori and H2O2 increase AP endonuclease-1/redox factor-1 expression in human gastric epithelial cells. AB - BACKGROUND & AIMS: Helicobacter pylori infection causes inflammation, accumulation of reactive oxygen species, and oxidative DNA damage in the gastric mucosa. Apurinic/apyrimidinic endonuclease-1 (APE-1)/redox factor-1 (Ref-1) repairs damaged DNA and reductively activates transcription factors, including activator protein-1. Considering that H. pylori generate reactive oxygen species and that reactive oxygen species modulate APE-1/Ref-1 in other cell types, we examined the effect of H. pylori, oxidative stress, and antioxidants on APE-1/Ref 1 expression in human gastric epithelial cells. METHODS: Human gastric epithelial cell lines or cells isolated from mucosal biopsy samples were stimulated with H. pylori, Campylobacter jejuni, and/or H 2 O 2 in the presence or absence of antioxidants. APE-1/Ref-1 expression was assayed by Western blot or reverse transcription polymerase chain reaction, and its cellular distribution was determined by using indirect conventional and confocal immunofluorescence. New protein synthesis was detected by [S 35 ]methionine labeling. APE-1/Ref-1 function was assessed by using a luciferase-linked reporter construct containing 3 activator protein 1 binding sites. RESULTS: APE-1/Ref-1 protein and messenger RNA were detected in resting gastric epithelial cells. APE-1/Ref-1 protein expression was increased after stimulation with H 2 O 2 or live cag pathogenicity island-bearing H. pylori, but not cag pathogenicity island-negative H. pylori or C. jejuni. H. pylori - or reactive oxygen species-mediated increases in APE-1/Ref 1 expression involved de novo protein synthesis that was inhibited by antioxidants. H. pylori or H 2 O 2 also induced nuclear accumulation of APE-1/Ref 1, and overexpression of APE-1/Ref-1 increased activator protein 1 binding activity. CONCLUSIONS: The data show that H. pylori or reactive oxygen species enhance APE-1/Ref-1 protein synthesis and nuclear accumulation in human gastric epithelial cells and implicate APE-1/Ref-1 in the modulation of the pathogenesis of H. pylori infection. PMID- 15362041 TI - Differing roles of protein kinase C-zeta in disruption of tight junction barrier by enteropathogenic and enterohemorrhagic Escherichia coli. AB - BACKGROUND & AIMS: Enteropathogenic Escherichia coli and enterohemorrhagic E. coli harbor highly homologous pathogenicity islands yet show key differences in their mechanisms of action. Both disrupt host intestinal epithelial tight junctions, but the effects of enteropathogenic E. coli are more profound than those of enterohemorrhagic E. coli. The basis for this is not understood. The atypical protein kinase C isoform, protein kinase C-zeta, associates with and regulates the tight junction complex. The aim of this study was to compare the role of protein kinase C-zeta in the disruption of tight junctions after infection with enteropathogenic E. coli and enterohemorrhagic E. coli. METHODS: Model intestinal epithelial monolayers infected by enteropathogenic E. coli or enterohemorrhagic E. coli were used for these studies. RESULTS: Neither bisindolylmaleimide nor Go6976, which block several protein kinase C isoforms but not protein kinase C-zeta, protected against the decrease in transepithelial electrical resistance after enteropathogenic E. coli infection. Rottlerin at concentrations that block novel and atypical isoforms, including protein kinase C zeta, significantly attenuated the decrease in transepithelial electrical resistance. The specific inhibitory peptide, myristoylated protein kinase C-zeta pseudosubstrate, also significantly decreased the enteropathogenic E. coli associated decrease in transepithelial electrical resistance and redistribution of tight junction proteins. In contrast to enteropathogenic E. coli, the level of protein kinase C-zeta enzyme activity stimulated by enterohemorrhagic E. coli was transient and minor, and protein kinase C-zeta inhibition had no effect on the decrease in transepithelial electrical resistance or the redistribution of occludin. CONCLUSIONS: The differential regulation of protein kinase C-zeta by enteropathogenic E. coli and enterohemorrhagic E. coli may in part explain the less profound effect of the latter on the barrier function of tight junctions. PMID- 15362042 TI - Immune stimulation of hepatic fibrogenesis by CD8 cells and attenuation by transgenic interleukin-10 from hepatocytes. AB - BACKGROUNDS & AIMS: Immunomodulatory cytokines, including interleukin-10 (IL-10), may mediate hepatic fibrosis. METHODS: We generated transgenic (TG) mice with hepatocyte expression of rat IL-10 (rIL-10) to assess its impact on lymphocyte subsets and activation of hepatic stellate cells following liver injury from carbon tetrachloride (CCl 4 ) or thioacetamide (TAA). RESULTS: Fibrosis was reduced in the TG animals in both models, which was not explained solely by differences in liver injury. By fluorescence-activated cell sorter (FACS), there were less CD4+ T cells in naive TG mice, and, following fibrosis induction, CD4+ T cells decreased only in wild-type (WT) mice, whereas increases in CD8+ T cells seen in WT animals were significantly attenuated in TG mice. Subtotal irradiation diminished fibrosis equally in both WT and TG groups, suggesting that rIL-10's antifibrotic effect was lymphocyte mediated. To assess the role of lymphocytes on stellate cell activation, either whole splenic lymphocytes, CD4+, or CD8+ T-cell subsets from WT animals with CCl 4 fibrosis were adoptively transferred to severe combined immunodeficiency (SCID) recipients, which led to stellate cell activation and fibrogenic stimulation as assessed by expression of transforming growth factor (TGF)-beta1 and collagen I messenger RNA (mRNA) and by immunoblot of alpha-smooth muscle actin. Moreover, serum aminotransferase levels and stellate cell activation mRNA were significantly higher among the CD8+ T-cell recipients. CONCLUSIONS: Transgenic expression of rIL-10 in liver leads to reduced fibrosis and alterations in liver lymphocyte subsets both in untreated liver and following fibrosis induction. In this model, fibrosis may be a CD8+ T cell-mediated disease that is attenuated by rIL-10. PMID- 15362043 TI - Trypsin mediates nociception via the proteinase-activated receptor 2: a potentially novel role in pancreatic pain. AB - BACKGROUND & AIMS: The pathogenesis of pain in pancreatitis remains poorly understood. We hypothesized that trypsin, a key inflammatory mediator in this condition, can also activate nociceptive neurons via the proteinase-activated receptor 2. METHODS: Double immunohistochemical staining of T8 to T12 dorsal root ganglia sections was performed with antibodies against proteinase-activated receptor 2 and vanilloid receptor 1, a marker for primary nociceptive neurons. In vivo nociceptive activity was measured by FOS immunoreactivity in thoracic spinal dorsal horn segments after intrapancreatic administration of proteinase-activated receptor 2 agonists. Pain behavior was assessed by visceromotor reflex activity in response to noxious stimulation of the pancreas with proteinase-activated receptor 2 agonists. RESULTS: Proteinase-activated receptor 2 was expressed by virtually all nociceptive neurons in thoracic dorsal root ganglia. Intraductal trypsin, in subinflammatory concentrations, activated spinal dorsal horn neurons in a dose-dependent manner, as measured by FOS expression. Both trypsin and a proteinase-activated receptor 2-specific peptide agonist induced a behavioral pain response when infused into the pancreatic duct of awake rats. Preinfusion of the pancreatic duct with proteinase-activated receptor 2-specific activating peptide desensitized the response to trypsin. CONCLUSIONS: Our findings suggest a novel proteinase-activated receptor 2-mediated role for trypsin in the pathogenesis of pancreatic pain and one that is independent of its inflammatory effect. PMID- 15362044 TI - Inducible histamine protects mice from P. acnes-primed and LPS-induced hepatitis through H2-receptor stimulation. AB - BACKGROUND & AIMS: Inducible histamine and histamine H2-receptors have been suggested to be involved in innate immune response. METHODS: We examined a functional role of inducible histamine in the protection against hepatic injury and lethality in Propionibacterium acnes -primed and lipopolysaccharide-induced hepatitis, using histidine decarboxylase knockout and H2-receptor knockout mice. RESULTS: Lipopolysaccharide challenge after Propionibacterium acnes priming increased histidine decarboxylase activity in the liver of wild-type mice, associated with a marked elevation of histamine turnover. Histidine decarboxylase like immunoreactivity was observed in CD68-positive Kupffer cells/macrophages. Treatment of wild-type mice with famotidine or ranitidine but not d chlorpheniramine augmented hepatic injury and inhibited the survival rate significantly. The same dose of Propionibacterium acnes and lipopolysaccharide induced severe hepatitis and high lethality in histidine decarboxylase knockout and H2-receptor knockout mice; the former were rescued by the subcutaneous injection of histamine. Immunohistochemical study supported the protective role of histamine against the apoptosis of hepatocytes. Histamine suppressed the expression of IL-18 and tumor necrosis factor alpha in the liver, leading to the reduced plasma levels of cytokines including IL-18, TNF-alpha, IL-12, IFN-gamma, and IL-6. CONCLUSIONS: These findings as a whole indicated that endogenously produced histamine in Kupffer cells/macrophages plays a very important role in preventing excessive innate immune response in endotoxin-induced fulminant hepatitis through the stimulation of H2-receptors. PMID- 15362045 TI - Calcium-dependent regulation of secretion in biliary epithelial cells: the role of apamin-sensitive SK channels. AB - BACKGROUND & AIMS: Increases in intracellular Ca 2+ are thought to complement cAMP in stimulating Cl - secretion in cholangiocytes, although the site(s) of action and channels involved are unknown. We have identified a Ca 2+ -activated K + channel (SK2) in biliary epithelium that is inhibited by apamin. The purpose of the present studies was to define the role of SK channels in Ca 2+ -dependent cholangiocyte secretion. METHODS: Studies were performed in human Mz-Cha-1 cells and normal rat cholangiocytes (NRC). Currents were measured by whole-cell patch clamp technique and transepithelial secretion by Ussing chamber. RESULTS: Ca 2+ dependent stimuli, including purinergic receptor stimulation, ionomycin, and increases in cell volume, each activated K + -selective currents with a linear IV relation and time-dependent inactivation. Currents were Ca 2+ dependent and were inhibited by apamin and by Ba 2+. In intact liver, immunoflourescence with an antibody to SK2 showed a prominent signal in cholangiocyte plasma membrane. To evaluate the functional significance, NRC monolayers were mounted in a Ussing chamber, and the short-circuit current ( I sc ) was measured. Exposure to ionomycin caused an increase in I sc 2-fold greater than that induced by cAMP. Both the basal and ionomycin-induced I sc were inhibited by basolateral Ba 2+, and approximately 58% of the basolateral K + current was apamin sensitive. CONCLUSIONS: These studies demonstrate that cholangiocytes exhibit robust Ca 2+ stimulated secretion significantly greater in magnitude than that stimulated by cAMP. SK2 plays an important role in mediating the increase in transepithelial secretion due to increases in intracellular Ca 2+. SK2 channels, therefore, may represent a target for pharmacologic modulation of bile flow. PMID- 15362046 TI - Role of phosphatidylinositol 3-kinase in the development of hepatocyte preconditioning. AB - BACKGROUND & AIMS: Ischemic preconditioning has been proved effective in reducing ischemia/reperfusion injury during liver surgery. However, the mechanisms involved are still poorly understood. Here, we have investigated the role of phosphatidylinositol 3-kinase (PI3K) in the signal pathway leading to hepatic preconditioning. METHODS: PI3K activation was evaluated in isolated rat hepatocytes preconditioned by 10-minute hypoxia followed by 10-minute reoxygenation. RESULTS: Hypoxic preconditioning stimulated phosphatidylinositol 3,4,5-triphosphate production and the phosphorylation of PKB/Akt, a downstream target of PI3K. Conversely, PI3K inhibition by wortmannin or LY294002 abolished hepatocyte tolerance against hypoxic damage induced by preconditioning. PI3K activation in preconditioned hepatocytes required the stimulation of adenosine A 2A receptors and was mimicked by adenosine A 2A receptors agonist CGS21680. In the cells treated with CGS21680, PI3K activation was prevented either by inhibiting adenylate cyclase and PKA with, respectively, 2,5-dideoxyadenosine and H89 or by blocking Galphai-protein and Src tyrosine kinase with, respectively, pertussis toxin and PP2. H89 also abolished the phosphorylation of adenosine A 2A receptors. However, the direct PKA activation by forskolin failed to stimulate PI3K. This suggested that PKA-phosphorylated adenosine A 2A receptors may activate PI3K by coupling it with Galphai-protein through Src. We also observed that, by impairing PI3K-mediated activation of phospholypase Cgamma (PLCgamma), wortmannin and LY294002 blocked the downstream transduction of preconditioning signals via protein kinase C (PKC) delta/ isozymes. CONCLUSIONS: PI3K is activated following hepatocyte hypoxic preconditioning by the combined stimulation of adenosine A 2A receptors, PKA, Galphai protein, and Src. By regulating PKC-/delta-dependent signals, PI3K can play a key role in the development of hepatic tolerance to hypoxia/reperfusion. PMID- 15362047 TI - High resolution analysis of cellular immune responses in resolved and persistent hepatitis C virus infection. AB - BACKGROUND & AIMS: Cellular immune responses are thought to play a key role in the resolution of primary HCV infection. Although it has been consistently shown that CD4+ T-cell responses are maintained in those with spontaneous resolution but lost in those with persistent infection, the role of CD8+ T-cell responses remains controversial. Previous studies have largely focused on limited HLA alleles and predefined CD8+ T-cell epitopes, and, thus, comprehensive studies remain to be performed. METHODS: To understand the composition of the immune response associated with spontaneous resolution, we comprehensively mapped CD8+ T cell responses in 20 HLA-diverse persons with resolved HCV infection, using HCV peptides spanning the entire genome. We analyzed the magnitude, breadth, function, and phenotype using ELISpot, class-I tetramers, intracellular cytokine staining, and cytolytic assays. We studied in parallel HCV-specific responses and viral sequence variation in persistent infection. RESULTS: Responses in individuals with resolved infection were strong and broad with robust proliferation in response to antigen. Responses in those persistently infected were rarely detected ex vivo and, when present, were narrowly directed and weak. However, they also proliferated in vitro. Dominant target epitopes differed among individuals in both cohorts, despite frequently shared HLA-alleles. CONCLUSIONS: These data indicate that persisting, strong CD8+ T-cell responses are observed in the majority of persons with resolved HCV infection and provide support for strategies to boost CD8+ T-cell responses for the prevention or treatment of HCV infection but also highlight the diversity of responses that may need to be elicited to provide protection. PMID- 15362048 TI - PTF1alpha/p48 transcription factor couples proliferation and differentiation in the exocrine pancreas [corrected]. AB - BACKGROUND & AIMS: The basic helix-loop-helix transcription factor pancreas specific transcription factor 1alpha (PTF1alpha)/p48 is critical for committing cells to a pancreatic fate and for the maintenance of the differentiated state in acinar cells. The aim was to analyze the ability of p48 to modulate cell proliferation, its relationship with cell differentiation, and the mechanisms involved therein. METHODS: Pancreatic and nonpancreatic cells were transfected with p48 cDNA, and the effects on cell proliferation were examined. The effects on cell cycle regulators were analyzed by Western blotting and RT-PCR; transient transfection assays were used to analyze promoter regulation. RESULTS: p48 Inhibited proliferation of acinar and nonacinar cells by inducing a delay in G1-S progression through the up-regulation of p21 CIP1/WAF1 and p27 KIP1 and the down regulation of cyclin D2. A 2-fold increase in p21 CIP1/WAF1 mRNA and in the activity of the p21 CIP1/WAF1 promoter was observed. The growth inhibition action of p48 was not associated with exocrine differentiation or with apoptosis. The antiproliferative effects were dependent on the COOH-terminal region of p48 and did not require the bHLH domain. Loss of p48 expression occurring during acinar to-ductal transitions, characteristic of chronic pancreatitis, was associated with an increase of cell proliferation in ductal complexes. CONCLUSIONS: The results indicate that p48 couples cell proliferation and cell differentiation in the exocrine pancreas, thus contributing to tissue homeostasis. These effects may play a role in the increased risk for pancreatic cancer associated with chronic pancreatitis. PMID- 15362049 TI - Ulcerative colitis with low-grade dysplasia. PMID- 15362050 TI - New insights into neurohormonal regulation of pancreatic secretion. AB - The existence of high- and low-affinity cholecystokinin (CCK)-A receptors on rodent pancreatic acini is well established. Until recently, CCK was believed to act directly on pancreatic acini to stimulate pancreatic secretion in both rodents and humans. However, conclusive evidence that human pancreatic acini lack functional CCK-A receptors has been presented. Despite substantial differences in rodent and human pancreatic physiology, CCK appears to act via vagal cholinergic pathways to mediate pancreatic secretion in both species. Structural and functional evidence suggests that CCK acts on vagal afferent fibers, which may explain how CCK doses that produce physiologic plasma CCK levels act via vagal cholinergic pathways to stimulate pancreatic secretion. Although most knowledge of vagal CCK-A receptors comes from research on rodents, physiologic studies suggest that this information is applicable to humans. In contrast to its effect on satiety, which is mediated by low-affinity vagal CCK-A receptors, CCK acts through high-affinity CCK-A receptors to evoke pancreatic secretion, suggesting that different affinity states of the vagal CCK receptors mediate different digestive functions. Vagal afferent pathways also transmit sensory information about the mechanical and physiochemical state of the digestive tract, mediated in part by serotonin, which, in turn, influences pancreatic secretion. A synergistic interaction between CCK and serotonin at the level of the nodose ganglia may explain the robust postprandial pancreatic secretion despite a modest postprandial increase in plasma CCK. Important physiologically, these findings not only explain discrepancies in previous in vivo vs. in vitro studies, but they revolutionize our current concept of the mechanism of CCK on pancreatic exocrine secretion. PMID- 15362051 TI - AGA future trends report: CT colonography. AB - BACKGROUND & AIMS: Computed tomographic colonography (CTC) was first described more than a decade ago. Recent advances in imaging hardware and software and results of clinical trials based on new methods for performing and interpreting images suggest that CTC may now be assessed as a method for colorectal cancer screening. METHODS: The Research Policy Committee of the American Gastroenterological Association assembled a task force to review the results of recent clinical trials and quantitative mathematical models pertaining to CTC. The goal of the task force was to assess the current knowledge about CTC and to evaluate the issues that will define its impact. RESULTS: Limitations in evaluating the current state of CTC technology include a wide variation in results of clinical trials. There are as yet insufficient data on the use of CTC in routine clinical practice. Limitations in the use of quantitative mathematical models make predictions based on such models of limited value. The cancer risk and therefore clinical importance of small colorectal polyps detected by CTC and/or nonpolypoid neoplasia not detected by CTC remains largely unknown. CONCLUSIONS: CTC is attractive as a colon imaging modality. It is therefore anticipated that CTC will have a significant impact on the practice of gastroenterology. However, the magnitude of the impact is currently unknown. Whether the ongoing implementation of CTC will increase or decrease the number of referrals for colonoscopy or shift the procedure from colorectal cancer screening to therapeutic interventions (e.g., polypectomy) is unknown at the present time. Multidisciplinary collaboration between gastroenterology and radiology to promote effective implementation and ongoing quality assurance will be important. PMID- 15362052 TI - The AGA and future trends in Gastroenterology: CT colonography. PMID- 15362053 TI - Confocal colonoscopy: more than skin deep. PMID- 15362054 TI - Screening for colorectal cancer: good, but not perfect. PMID- 15362055 TI - The efficacy of azathioprine and 6-mercaptopurine for the prevention of postoperative recurrence in patients with Crohn's disease remains uncertain. PMID- 15362056 TI - A CLA's act: feeding away inflammation. PMID- 15362057 TI - Inflammation, damage repair, immune cells, and liver fibrosis: specific or nonspecific, this is the question. PMID- 15362058 TI - Mesalamine in IBD: multiple hats? PMID- 15362060 TI - Does an aspirin a day keep pancreas cancer away? PMID- 15362061 TI - Lysine as a serotonin receptor antagonist: using the diet to modulate gut function. PMID- 15362062 TI - What a difference a datum makes. PMID- 15362064 TI - An increasing incidence of cholangiocarcinoma: why? PMID- 15362069 TI - The role of nitric oxide in gastrointestinal health and disease. PMID- 15362070 TI - Should GOV1 be treated as for esophageal varices? PMID- 15362072 TI - Discussion on prophylactic antibiotic treatment in patients with predicted severe pancreatitis: a placebo-controlled, double-blind trial. PMID- 15362074 TI - Hemorrhoids and hemorrhoidectomies. PMID- 15362075 TI - Risk assessment of endogenous formation of carcinogenic N-nitroso compounds in response to intake of NO-aspirin. PMID- 15362080 TI - Pharmionics: research on what patients do with prescription drugs. PMID- 15362081 TI - Nonadherence as a predictor of antidiabetic drug therapy intensification (augmentation). AB - PURPOSE: To determine if nonadherence with antidiabetic drug therapy is predictive of subsequent antidiabetic drug therapy intensification. METHODS: We conducted a retrospective cohort study examining retail pharmacy dispensings of sulfonylureas or metformin to 1067 patients having diabetes. Patients that did not receive a sufficient quantity of medication to cover at least 80% of days during the evaluation period were classified as nonadherent. Outcomes identified were increase in the dose of antidiabetic medication utilized, the addition of a second antidiabetic agent to the regimen or either. RESULTS: Among users of sulfonylurea monotherapy, those classified as nonadherent were 45% more likely to intensify therapy in subsequent months as compared with those classified as adherent (age-adjusted odds ratio (OR) 1.45; 95% confidence interval (CI) 1.06 2.00). This finding was largely driven by observed increases in dosage, which were more likely among patients classified as nonadherent (age-adjusted OR 1.48, 95%CI 1.07-2.05). Nonadherence was not found to be predictive of the subsequent addition of a second antidiabetic agent (OR 1.02; 95%CI 0.64-1.63). Overall findings were similar for the smaller sample of patients receiving metformin monotherapy, though observed differences did not achieve statistical significance. CONCLUSIONS: Patients who were poorly adherent to oral antidiabetic drug therapy more frequently experienced an increase in the dose of medication prescribed, as compared to patients that were classified as adherent. This finding underscores the need for prescribers to consider nonadherence as a root cause when patients fail to achieve therapeutic goals. PMID- 15362082 TI - The role of scientific evidence of risks and benefits in determining risk management policies for medications. AB - Recent changes in the regulatory environment have called attention to the need for and potential benefits of greater and more detailed evidence to inform decisions based on the risk-benefit profile of medications. Nevertheless, access to potentially beneficial therapies continues to be impeded by a lack of sufficient information that could help optimize benefits and minimize risks of treatments for patients. Over-reliance on pre-marketing clinical trials and the FDA's spontaneous reporting adverse event system to support regulatory decisions has sustained an information void. Clinical trials are the gold standard for demonstrating efficacy, but they cannot fully predict safety when drugs are used in the real world. Spontaneous reporting can identify new signals, but cannot quantify those signals or place them in appropriate clinical context. In the face of new safety signals, absence of better information on how medications are used and how they perform in the real world setting, regulators are often limited to either continuing drug marketing without significant changes or withdrawing a medication from the market. Experience shows that information collected proactively, to better understand the background risks associated with the underlying disease and to better quantify the product risks, can influence these decisions to include a wider range of options regarding a product's availability, labeling and additional risk management strategies. This article presents several case studies of medications, including those in which insufficient data were available to address important safety signals and decisions were made to withdraw products, as well as those in which epidemiologic data were available to provide reassurance of product safety and allow continued product use, even though some may be marketed with additional risk management programs. More extensive and earlier epidemiologic assessment of risks and benefits of new products will create a new standard of evidence for industry and regulators and is likely to result in more effective and balanced regulatory actions, thereby affording better care for patients. PMID- 15362083 TI - Drug use among fathers around time of conception: two register based surveys from Denmark and The Netherlands. AB - STUDY OBJECTIVE: Despite the increasing attention for the role of paternal exposures around the period of conception, there is no factual information about drug utilisation of fathers. Therefore, the aim of this study was to describe the drugs dispensed to fathers around conception, using pharmacy dispensing data of community pharmacies in Denmark and The Netherlands. DESIGN AND SETTING: Using pharmacy dispensing data from the Pharmaco-epidemiological Prescription Database of North Jutland in Denmark and the InterAction database in The Netherlands, we examined the prescriptions reimbursed in the half year before conception of 56,735 Danish fathers from 1991 to 2000, and 5859 Dutch fathers from 1995 to 2000. MAIN RESULTS: One third of all fathers had taken up prescriptions for at least one drug in the half year before conception, both in Denmark and in The Netherlands. In the majority of fathers only one type of drug was dispensed, but in both countries at least 5% of all fathers had redeemed three or more types of drugs. The main drugs purchased by fathers in Denmark and The Netherlands were antibiotics (14.3 and 6.3% of all fathers, respectively), analgesics (6.1 and 7.6%), antihistamines (2.0 and 2.0%) and anti-ulcer drugs (1.6 and 2.5%). CONCLUSION: A large proportion of fathers used drugs around the time of conception. This finding emphasises the importance of safety information on therapeutic drugs with respect to potential paternal teratogenicity. PMID- 15362084 TI - Post-marketing surveillance of buprenorphine. AB - PURPOSE: This study was undertaken to evaluate the adverse consequences of recently introduced higher strength (0.4 and 2.0 mg per tablet) buprenorphine in Indian market. Buprenorphine, a partial opiate agonist and antagonist, is an emerging alternative to methadone as an agent for long-term treatment of opiate dependence. METHODS: The current investigation was conducted through a multi centric post-marketing surveillance (PMS) study using a structured performa from patients receiving buprenorphine as routine therapy from de-addiction centres. Evaluation included subjective and objective assessments and recording of adverse events. RESULTS: Of the 5551 observations from ten centres, common subjective symptoms were generalised weakness (48.9%), sense of high (euphoria) (44.5%), muscle aches (39.5%) and relief from pain (37.2%). About 5% observations recorded systolic hypertension. Among 55 subjects where laboratory tests were conducted, 12 showed raised levels of AST ad 9 had elevated ALT. Twelve adverse events reported included seizure, epistaxis, panic attacks, constipation and dyspnoea. Significant relation was seen between duration of use and time since last dose, and total number of subjective symptoms reported. CONCLUSIONS: Majority of the adverse effects could be understood as either effects related to intoxication or withdrawal from agonists. PMID- 15362085 TI - Abstracts on drug utilization research at seven International Conferences on Pharmacoepidemiology (ICPE): 1996-2002. PMID- 15362086 TI - Use of time-dependent measures to estimate benefits of beta-blockers after myocardial infarction. AB - PURPOSE: To estimate the reduction in all-cause mortality conferred by beta blockers in a population-based cohort of elderly survivors of myocardial infarction during the year following hospital discharge. METHODS: A dynamic retrospective cohort was assembled from persons aged 66 years and over surviving myocardial infarction in Quebec between 1990 and 1993. Information on hospitalizations was linked to medication and physician claims, demographic characteristics and vital status. Subjects prescribed beta-blockers at hospital discharge had fewer comorbid medical conditions, less pre-existing cardiovascular disease and less severe infarcts. To control for these differences, analyzes were restricted to subjects receiving at least one beta-blocker and mortality was compared between periods with and without beta-blocker exposure using Cox proportional hazard models. RESULTS: Among 14,547 survivors of myocardial infarction, 41% were dispensed at least one beta-blocker. Among those subjects, the risk of dying during periods of beta-blocker use was reduced 40% (hazard ratio = 0.6; 95% CI: 0.5, 0.7). CONCLUSION: Confounding by indication threatens the validity of observational studies of intended effects of medications. For elderly survivors of myocardial infarction, the estimated benefit of beta blockers from observational studies is greater than the estimate from randomized trials. Greater benefits do not seem to be an artifact arising from systematically prescribing beta-blockers to subjects with better prognosis. Reducing confounding by indication can enhance the validity of observational studies of medications and widen research applications of administrative health databases. While the actual benefits of medications are never truly known these studies can provide a credible range that brackets the truth. PMID- 15362087 TI - Evaluation of drug prescription at the Geneva prison's outpatient service in comparison to an urban outpatient medical service. AB - PURPOSE: To analyse non-psychotropic drug prescription in a prison outpatient clinic in comparison with an urban medical outpatient service. METHODS: Comparative study during 3 weeks at Geneva: prison outpatient service and medical policlinic (MP) of the University Hospital. RESULTS: The most often prescribed non-psychotropic drugs at the Geneva prison were systemic analgesics (mostly non steroidal anti-inflammatory drugs (NSAID) and paracetamol), dermatologicals, systemic anti-infectives and drugs for the gastrointestinal system. For most types of non-psychotropic drugs, frequency of prescription as well as the prescribing patterns were similar in the prison ambulatory service and the urban MP. Dermatologicals were prescribed more frequently at the prison than at the MP. Analgetics have been prescribed mainly for osteoarticulary reasons, especially low back pain at the MP, and for traumatism and headache at the prison. CONCLUSIONS: The higher frequency of dermatological prescriptions could be due to prison environmental factors. We do not have arguments for any overprescription of analgesics or other non-psychotropic drugs. The data did not show any prescription or co-prescription of several substances that violated clinical guidelines. PMID- 15362088 TI - Antimicrobials for the treatment of urinary tract infection in pregnancy: practices in southern India. AB - PURPOSE: Although urinary tract infection (UTI) in pregnancy is a common medical problem, very little is known about the susceptibility patterns of bacteria causing UTI and the prescription practices for UTI in pregnancy in low-income countries. Studies were, therefore, carried out in southern India to document the susceptibility patterns and to compare these with prescription practices. METHODS: Data on susceptibility patterns of bacteria isolated in significant counts from urine of pregnant women suspected to have urinary infection were collected. Prescribing patterns for these infections were ascertained using a questionnaire. RESULTS: About 90% of E. coli causing UTI is still susceptible to nitrofurantoin, a relatively inexpensive and safe drug. However, less than 25% of doctors used it for treatment of cystitis. Cephalosporins were most commonly used in hospital practice for the treatment of UTI. Amoxycillin was being used widely to treat UTI in pregnancy in spite of high prevalence of resistance. There were wide variations in the duration of therapy and use of prophylaxis. CONCLUSIONS: Our findings indicate an urgent need for proper guidelines, dissemination of information to practitioners and supervision of antimicrobial usage in low income countries like India. Irrational and unnecessary drug use can be expensive and harmful. PMID- 15362089 TI - Utilization of parenteral anti-infective agents in the medical emergency unit of a tertiary care hospital: an observational study. AB - A pharmacy based prescription audit was undertaken in the medical emergency unit of a tertiary care hospital to determine the frequency of prescribing of parenteral anti-infective agents. During the study period, 885 patients were screened. The analysis was done for the number of parenteral anti-microbials in each prescription frequency of individual drug prescribe number and dose unit (DDD), frequency of age and sex, site of infection and daily cost incurred by the patient. It was found that 400 patients (45.2%) received parenteral anti infective agents. Cephalosporins, aminoglycosides and metronidazole accounted for about 70% of total antimicrobial use. More than 50% of patients had culture sensitivity reports available. The mean (standard deviation, SD) daily cost of antibiotic was USD 3.8 (7.7), median; range 2 (0.1-85.7). Two anti-microbials per prescription were indicated in most of the patients (43.88). In conclusion we have provided an overview of parenteral anti-infective use in medical emergency, which may serve as a basis for intervention and improvement in prescribing pattern of parenteral anti-microbials. PMID- 15362090 TI - Psychotropic drugs and fatal pulmonary embolism: a comment. PMID- 15362091 TI - Proceedings of the EMBO Workshop on Biological Implications of Macromolecular Crowding. June 14-18, 2003. Avila, Spain. PMID- 15362092 TI - A potential role for isothermal calorimetry in studies of the effects of thermodynamic non-ideality in enzyme-catalyzed reactions. AB - Attention is drawn to the feasibility of using isothermal calorimetry for the characterization of enzyme reactions under conditions bearing greater relevance to the crowded biological environment, where kinetic parameters are likely to differ significantly from those obtained by classical enzyme kinetic studies in dilute solution. An outline of the application of isothermal calorimetry to the determination of enzyme kinetic parameters is followed by considerations of the nature and consequences of crowding effects in enzyme catalysis. Some of those effects of thermodynamic non-ideality are then illustrated by means of experimental results from calorimetric studies of the effect of molecular crowding on the kinetics of catalysis by rabbit muscle pyruvate kinase. This review concludes with a discussion of the potential of isothermal calorimetry for the experimental determination of kinetic parameters for enzymes either in biological environments or at least in media that should provide reasonable approximations of the crowded conditions encountered in vivo. PMID- 15362093 TI - Non-ideal tracer sedimentation equilibrium: a powerful tool for the characterization of macromolecular interactions in crowded solutions. AB - Non-ideal tracer sedimentation equilibrium is a technique devised to quantify the effect of high concentrations of unrelated macromolecules on the self- or hetero associations of dilute macromolecules. Principles and experimental techniques are reviewed, and previous experimental work summarized. A new analysis of experimental data is presented that requires no a priori assumptions regarding the nature of weak repulsive interactions between solute species and the concentrated (crowding) species. PMID- 15362094 TI - Protein folding and binding in confined spaces and in crowded solutions. AB - Simple theoretical models are presented to illustrate the effects of spatial confinement and macromolecular crowding on the equilibria and rates of protein folding and binding. Confinement is expected to significantly stabilize the folded state, but for crowding only a marginal effect on protein stability is expected. In confinement the unfolded chain is restricted to a cage but in crowding the unfolded chain may explore different interstitial voids. Because confinement and crowding eliminate the more expanded conformations of the unfolded state, folding from the compact unfolded state is expected to speed up. Crowding will shift the binding equilibrium of proteins toward the bound state. The significant slowing down in protein diffusion by crowding, perhaps beneficial for chaperonin action, could result in a decrease in protein binding rates. PMID- 15362095 TI - Crowding-induced organization in cells: spontaneous alignment and sorting of filaments with physiological control points. AB - Under sufficiently crowded conditions, elongated particles spontaneously align along a common axis and separate from particles with dissimilar packing parameters. Clarifying the relevance of these entropy-driven phenomena to intact cells has required the development of theoretical approaches that tractably take into account daunting physiological complexities including the extreme crowding of the cytosol, the complex mixture of macromolecules present, the process of filament self-assembly, and the characteristic widths, flexibilities and charges of filaments formed by different proteins. This review summarizes the approaches taken, including their validation by observations of simpler systems, and the insights that have been gained into the means by which cells can modulate and capitalize upon spontaneous ordering. PMID- 15362096 TI - Crowding in extremophiles: linkage between solvation and weak protein-protein interactions, stability and dynamics, provides insight into molecular adaptation. AB - The study of the molecular adaptation of microorganisms to extreme environments (solvent, temperature, etc.) has provided tools to investigate the complex relationships between protein-solvent and protein-protein interactions, protein stability and protein dynamics, and how they are modulated by the crowded environment of the cell. We have evaluated protein-solvent and protein-protein interactions by solution experiments (analytical ultracentrifugation, small angle neutron and X-ray scattering, density) and crystallography, and protein dynamics by energy resolved neutron scattering. This review concerns work from our laboratory on (i) proteins from extreme halophilic Archaea, and (ii) psychrophile, mesophile, thermophile and hyperthermophile bacterial cells. PMID- 15362097 TI - Bacteriophage O29 protein p6: an architectural protein involved in genome organization, replication and control of transcription. AB - Protein p6 of B. subtilis bacteriophage O29 binds to DNA forming a nucleoprotein complex in which the DNA wraps a protein core forming a right-handed superhelix, therefore restraining positive supercoiling and compacting the DNA. The protein does not specifically recognize a nucleotide sequence but rather a structural feature and it binds as a dimer through the minor groove. Protein p6 is in a monomer-dimer equilibrium that shifts to higher-order structures at a concentration of about 1 mM. These structures are probably present in vivo as the intracellular concentration of p6 is estimated to be in this range, and in fact the effective concentration should be still higher due to the macromolecular crowding. The p6 oligomers show an elongated shape compatible with a helical structure reminiscent of the superhelical DNA of the nucleoprotein complex, therefore it was proposed that protein p6 forms a scaffold on which the DNA folds. Since protein p6 is very abundant in infected cells, enough to bind the entire viral progeny, it was proposed to have an architectural role organizing and compacting the viral genome. It has been demonstrated that protein p6 binds in vivo to most, if not all, the O29 genome, although with different affinity, the highest one corresponding to the genome ends. Binding to plasmidic DNA was much lower, although it increased dramatically when the negative superhelicity was decreased. Hence, protein p6 binding specificity for O29 DNA is based on supercoiling, providing that the O29 genome, although topologically constrained, has a negative superhelicity lower than that of plasmid DNA. The formation of the nucleoprotein complex has functional implications in DNA replication and the control of transcription. It activates the initiation of replication that occurs at the genome ends for which the binding affinity is highest. It represses early transcription from promoter C2, and, together with protein p4, it represses transcription from promoters A2b and A2c and activates late transcription from promoter A3; therefore, protein p6 is involved in the early to late transcription switch. PMID- 15362098 TI - Macromolecular crowding in biological systems: hydrodynamics and NMR methods. AB - Most biologically relevant environments involve highly concentrated macromolecular solutions and most biological processes involve macromolecules that diffuse and interact with other macromolecules. Macromolecular crowding is a general phenomenon that strongly affects the transport properties of macromolecules (rotational and translational diffusion) as well as the position of their equilibria. NMR methods can provide information on molecular interactions, as well as on translational and rotational diffusion. In fact, rotational diffusion, through its determinant role in NMR relaxation, places a practical limit on the systems that can be studied by NMR. While in dilute solutions of non-aggregating macromolecules this limit is set by macromolecular size, in crowded solutions excluded volume effects can have a strong effect on the observed diffusion rates. Hydrodynamic theory offers some insight into the magnitude of crowding effects on NMR observable parameters. PMID- 15362099 TI - Fluorescence anisotropy as a probe to study tracer proteins in crowded solutions. AB - Fluorescence anisotropy has been widely used to study the dynamics and interactions of biomolecules in diluted solutions. Comparable studies on single tracer macromolecules at the cellular level are now feasible because of the recent development of non-invasive fluorescence markers, like the growing family of the green fluorescence proteins (GFPs), and the advances in time-resolved fluorescence microscopy instrumentation. The interpretation of fluorescence polarization data in terms of dynamics and biological function of the macromolecular complexes in these physiological environments requires a deep understanding of the tracer rotational diffusion in such complex media. In this work we have studied the rotational diffusion of a tracer protein, apomyoglobin labeled with 1-anilino-8-naphthalene sulfonate, in crowded solutions of an unrelated protein, ribonuclease A. We have evaluated the deviation of the different tracer rotational motions from the Stokes-Einstein-Debye diffusion behavior, and its relation to the properties of the transient molecular cavities where the tracer is rotating in the fluorescence lifetime window. Finally, we have analyzed the application of fluorescence polarization methods to determine the apparent equilibrium constants of homo and hetero-associations of macromolecules in crowded conditions. PMID- 15362100 TI - Crowding effects in binary mixtures of rod-like and spherical particles. AB - Crowding effects relevant to the phase stability of binary mixtures of rod-like and spherical particles are investigated by means of Monte Carlo simulations in the isobaric NPT ensemble. The two types of particles are represented, respectively, by freely rotating hard spherocylinders of a moderate aspect ratio (L/sigma = 5) and hard spheres of the same diameter sigma. Molar fractions of spheres ranging xHS = 0.00-0.37 are considered with the aim of characterizing the crowding effects on the liquid crystal phases of the hard spherocylinder fluid induced by the spherical component as depleting agent. We find that the addition of the spherical crowder is beneficial for the stabilization of the layers of the rod-like particles characteristic of the smectic phase. On the contrary, the addition of spheres has a negative impact upon the stability of the nematic phase, where the rod-like particles tend to align collectively parallel to each other. Interestingly, the spheres tend to arrange forming rod-like clusters in the nematic phase and lamellar structures in the smectic phase, which is compensated by the entropy gained by the spherocylinder particles in each phase. The main results are in qualitative agreement with recent experimental and theoretical studies and serve to test the prediction of current equations of state for these types of binary mixtures. PMID- 15362101 TI - Effect of molecular crowding on self-association of phosphorylase kinase and its interaction with phosphorylase b and glycogen. AB - Self-association of phosphorylase kinase (PhK) and its interaction with glycogen (M=5500 kDa) and phosphorylase b (Phb) has been studied using analytical ultracentrifugation and turbidimetry under the conditions of molecular crowding arising from the presence of high concentrations of osmolytes. In accordance with the predictions of the molecular crowding theory, trimethylamine N-oxide (TMAO) and betaine greatly favor self-association of PhK induced by Mg2+ and Ca2+ and PhK interaction with glycogen. In contrast, proline suppresses these processes, probably, due to its specific interaction with PhK. All osmolytes tested prevented the complex formation between PhK and its physiological substrate, Phb. The specific interactions of PhK and Phb with glycogen, in the living cell, presumably is a factor allowing the negative effect of crowding on the recognition of Phb by PhK to be overcome. PMID- 15362102 TI - Studies of effects of macromolecular crowding and confinement on protein folding and protein stability. AB - In cells, proteins execute specific tasks in crowded environments; these environments influence their stability and dynamics. Similarly, for an enzyme molecule encapsulated in an inorganic cavity as in biosensors or biocatalysts, confinement or excluded volume plays an important role in its stability and dynamics. In this article we present results of our experimental and theoretical investigations of the confinement and macromolecular crowding effects on protein. On the experimental side we study the stability of encapsulated cytochrome c against unfolding induced by the presence of denaturants, such as urea. Results show that, as the pore size in which protein is trapped is reduced, protein shows higher stability against denaturant-induced unfolding. On the theoretical side, after reviewing our previous study of the confinement effects on the equilibrium and dynamic properties of protein using a minimalist (two-dimensional lattice, Monte Carlo, Brownian dynamics) model, we have extended the model so that the effects of macromolecular crowding on such properties can be studied. Our simulations show that both folding and unfolding times increase with the number of crowders in solution, however, the equilibrium constant is affected such that the equilibrium is shifted towards the folded state. Furthermore, our results show that, for a fixed number of crowders as the size of crowder (or excluded volume) increases, the average size of protein at equilibrium decreases. PMID- 15362103 TI - Molecular crowding reduces to a similar extent the diffusion of small solutes and macromolecules: measurement by fluorescence correlation spectroscopy. AB - Aqueous environments in living cells are crowded, with up to >50 wt% small and macromolecule-size solutes. We investigated quantitatively one important consequence of molecular crowding--reduced diffusion of biologically important solutes. Fluorescence correlation spectroscopy (FCS) was used to measure the diffusion of a series of fluorescent small solutes and macromolecules. In water, diffusion coefficients (D(o)w) were (in cm2/s x 10(-8)): rhodamine green (270), albumin (52), dextrans (75, 10 kDa; 10, 500 kDa), double-stranded DNAs (96, 20 bp; 10, 1 kb; 3.4, 4.5 kb) and polystyrene nanospheres (5.4, 20 nm diameter; 2.3, 100 nm). Aqueous-phase diffusion (Dw) in solutions crowded with Ficoll-70 (0-60 wt%) was reduced by up to 650-fold in an exponential manner: Dw = D(o)w exp ( [C]/[C]exp), where [C]exp is the concentration (in wt%) of crowding agent reducing D(o)w by 63%. FCS data for all solutes and Ficoll-70 concentrations fitted well to a model of single-component, simple (non-anomalous) diffusion. Interestingly [C]exp were nearly identical (11+/-2 wt%, SD) for diffusion of the very different types of macromolecules in Ficoll-70 solutions. However, [C]exp was dependent on the nature of the crowding agent: for example, [C]exp for diffusion of rhodamine green was 30 wt% for glycerol and 16 wt% for 500 kDa dextran. Our results indicate that molecular crowding can greatly reduce aqueous phase diffusion of biologically important macromolecules, and demonstrate a previously unrecognized insensitivity of crowding effects on the size and characteristics of the diffusing species. PMID- 15362104 TI - Applications of time-resolved resonance energy transfer measurements in studies of the molecular crowding effect. AB - The native structures of many globular proteins are only weakly stabilized and form in solution ensembles of multiple conformers. The energy differences between the conformers are assumed to be small. This is the case of flexible multidomain proteins where domain motions were observed. High concentrations of inert macrosolute, which create a crowded or confined environment, can cause shifts of the distribution of the conformers of such proteins towards the more compact structures. This effect may also promote compact structures in partially folded proteins. Time-resolved dynamic non-radiative excitation energy transfer (tr-RET) is suitable for detection of either subtle or major changes in distributions of intramolecular distances in protein molecules in solutions. Two experiments were performed which demonstrated the applicability of tr-RET for detection of the effect of macrosolutes on the conformational ensembles of flexible states of protein molecules. The distribution of distances between residues 203 and 169 in the CORE domain of E. coli adenylate kinase (AK) in the denatured state was determined in the presence of high concentrations of dextran 40. A significant shift of the mean of the distribution was observed without reduction of its width. This was interpreted as a shift to compact structure without change of the degree of disorder of the chain. In a second experiment the distribution of the distance between residues 55 and 169 in AK, which spans the cleft between the CORE and the AMPbind domains, was monitored. No clear effect of high concentrations of dextran 40 was found. These experiments show the strength of the application of tr-RET in investigation of changes in the sub-states of flexible conformations of globular protein. Networks of pairs of labeled sites can be prepared and tr-RET experiments can be performed in order to search for the segments of the protein molecules, which respond to the presence of inert macromolecules in their environment. PMID- 15362105 TI - The effect of macromolecular crowding on protein aggregation and amyloid fibril formation. AB - Macromolecular crowding is expected to have several significant effects on protein aggregation; the major effects will be those due to excluded volume and increased viscosity. In this report we summarize data demonstrating that macromolecular crowding may lead to a dramatic acceleration in the rate of protein aggregation and formation of amyloid fibrils, using the protein alpha synuclein. The aggregation of alpha-synuclein has been implicated as a critical factor in development of Parkinson's disease. Various types of polymers, from neutral polyethylene glycols and polysaccharides (Ficolls, dextrans) to inert proteins, are shown to accelerate alpha-synuclein fibrillation. The stimulation of fibrillation increases with increasing length of polymer, as well as increasing polymer concentration. At lower polymer concentrations (typically up to approximately 100 mg/ml) the major effect is ascribed to excluded volume, whereas at higher polymer concentrations evidence of opposing viscosity effects become apparent. Pesticides and metals, which are linked to increased risk of Parkinson's disease by epidemiological studies, are shown to accelerate alpha synuclein fibrillation under conditions of molecular crowding. PMID- 15362106 TI - Chaperonin function--effects of crowding and confinement. AB - Chaperonins assist in the acquisition of native protein structure in the cell by providing a shielded environment for a folding polypeptide chain, generated by the interior surface of their cylindrical structure. The folding chain is isolated from the highly crowded cytoplasm, but at the same time confined within the chaperonin folding cage. Both confinement and macromolecular crowding can affect folding kinetics and yields, the modus operandi of chaperonins and their interaction with their proteges. Recent experimental data, as well as computer simulations, provide increasing evidence that the particular physico-chemical conditions prevailing in the cellular interior have to be taken into account when trying to unravel the processes of cellular protein folding. PMID- 15362107 TI - Cell volume and the regulation of apoptotic cell death. AB - Apoptosis is a physiological mechanism allowing for the removal of abundant or potentially harmful cells. The hallmarks of apoptosis include degradation of cellular DNA, exposure of phosphatidylserine at the outer leaflet of the cell membrane and cell shrinkage. Phosphatidylserine exposure favours adhesion to macrophages with subsequent phagocytosis of the shrunken apoptotic particles. The interaction of cell volume regulatory mechanisms and apoptosis is illustrated in two different model systems, i.e. (a) lymphocyte apoptosis following stimulation of CD95 receptor and (b) erythrocyte apoptosis upon cell shrinkage. (a) Triggering of CD95 in Jurkat T lymphocytes is paralleled by activation of cell volume regulatory Cl- channels, inhibition of the Na+/H+ exchanger and osmolyte release. The latter coincides with cell shrinkage, DNA fragmentation and phosphatidylserine exposure. CD95 stimulation leads to early inhibition of the voltage gated K+ channel Kv1.3, which may contribute to the inhibition of the Ca2+ release activated Ca2+ channel I(CRAC). (b) Osmotic shock of erythrocytes activates a cell volume regulatory cation conductance allowing the entry not only of Na+ but of Ca2+ as well. Increased cytosolic Ca2+ stimulates a scramblase which disrupts the phosphatidylserine asymmetry of the cell membrane, leading to phosphatidylserine exposure. The cation conductance is further activated by oxidative stress and energy depletion and inhibited by Cl-. Shrinkage of erythrocytes stimulates in addition a sphingomyelinase with subsequent formation of ceramide which potentiates the effect of cytosolic Ca2+ on phosphatidylserine. In conclusion, cell volume-sensitive mechanisms participate in the triggering of apoptosis following receptor stimulation or cell injury. PMID- 15362108 TI - Genomic channeling in bacterial cell division. AB - The bacterial dcw cluster is a group of genes involved in cell division and peptidoglycan synthesis. Comparison of the cluster across several bacterial genomes shows that its gene content and its gene order are conserved in distant bacterial lineages and, moreover, that, being most conserved in rod-shaped bacteria, the degree of conservation relates to bacterial morphology. We propose a model in which the selective pressure to maintain the cluster arises from the need to efficiently coordinate the processes of elongation and septation in rod shaped bacteria. Gene order in the dcw cluster would be conserved as a result of mechanisms comprising: (i) a limited amount of peptidoglycan precursors required both for septation and elongation of the wall; (ii) co-translational assembly of the protein complexes involved in cell division and in the synthesis of the peptidoglycan precursors; and (iii) alternation in the cellular localization of the assembled complexes to participate either in the synthesis of the septal peptidoglycan and division, or in the synthesis of the lateral wall. The name genomic channeling is proposed for this model as it involves a genomic arrangement that could facilitate the assembly of specific protein complexes and their subsequent conveyance to specific locations in the crowded cytoplasm and the envelope. PMID- 15362109 TI - Large changes in cytoplasmic biopolymer concentration with osmolality indicate that macromolecular crowding may regulate protein-DNA interactions and growth rate in osmotically stressed Escherichia coli K-12. AB - From determination of amounts and concentrations of biopolymers and solutes in the cytoplasm of Escherichia coli, we are obtaining information needed to assess the effect of macromolecular crowding on cytoplasmic properties and processes of osmotically stressed bacteria. We observe that growth rate, and the amount of cytoplasmic water decrease and cytoplasmic concentrations of biopolymers and K+, increase with increasing osmolality, even for cells grown in the presence of osmoprotectants like glycine betaine. We observe general correlations between the amount of cytoplasmic water, growth rate and cytoplasmic K+ concentration in osmotically stressed cells grown both with and without osmoprotectants. To explain these correlations, we propose that crowding increases with increasing growth osmolality, which in turn buffers the binding of proteins to nucleic acids against changes in cytoplasmic K+ concentration and (by affecting biopolymer diffusion rates and/or assembly equilibria) is a determinant of growth rate of osmotically stressed cells. Changes in biopolymer concentration and crowding may also explain the increase of the activity coefficient of cytoplasmic water with increasing osmolality of growth in E. coli. PMID- 15362110 TI - Crowding and the polymerization of sickle hemoglobin. AB - Under physiological conditions, sickle hemoglobin, a natural mutant of human hemoglobin A with a surface hydrophobic valine in place of a negatively charged glutamic acid, polymerizes at high volume occupancy. Equilibrium solubility of sickle hemoglobin entails activity coefficients that can approach 10(3) at high concentrations. Polymerization occurs by homogeneous and heterogeneous nucleation mechanisms, which are both profoundly sensitive to crowding; homogeneous nucleation rates for example are enhanced by 10(10) when the initial concentration is augmented by 50% non-polymerizing hemoglobin. A molecular description of the reaction therefore entails substantial corrections for molecular crowding which are all very accurately described by excluded volume corrections, treating hemoglobin as a hard sphere with volume consistent with the molecular structure of the molecule, and involving no further adjustable parameters. These effects and the descriptions that rationalize this behavior are described. PMID- 15362111 TI - Neutron scattering measurements of intact cells show changes after heat shock consistent with an increase in molecular crowding. AB - Molecular crowding has been shown to be important in many cellular processes. The crowded environment in the cell results in a significant proportion of the cellular water being in contact with macromolecules such as proteins and DNA. These interfacial water molecules show a reduced dynamic motion that has been observed with isolated macromolecules using several biophysical techniques. Previously we investigated the inelastic neutron scattering properties of water closely associated with isolated biomolecules, and showed that interfacial water is strongly perturbed, as judged by its energy transfer spectrum. Here we have probed living cells using inelastic and quasielastic neutron scattering. We have found that mild heat stress ('heat shock'), which causes some proteins to become unfolded in the cell, results in changes in the inelastic neutron scattering in the librational region (45-130 meV). Heat shock also causes a narrowing of the quasielastic scattering peak. These changes can be understood in terms of an increase in the proportion of interfacial water molecules, and a net reduction in proton dynamics. PMID- 15362112 TI - Green fluorescent protein fusions to Arabidopsis fimbrin 1 for spatio-temporal imaging of F-actin dynamics in roots. AB - The visualization of green fluorescent protein (GFP) fusions with microtubule or actin filament (F-actin) binding proteins has provided new insights into the function of the cytoskeleton during plant development. For studies on actin, GFP fusions to talin have been the most generally used reporters. Although GFP-Talin has allowed in vivo F-actin imaging in a variety of plant cells, its utility in monitoring F-actin in stably transformed plants is limited particularly in developing roots where interesting actin dependent cell processes are occurring. In this study, we created a variety of GFP fusions to Arabidopsis Fimbrin 1 (AtFim1) to explore their utility for in vivo F-actin imaging in root cells and to better understand the actin binding properties of AtFim1 in living plant cells. Translational fusions of GFP to full-length AtFim1 or to some truncated variants of AtFim1 showed filamentous labeling in transient expression assays. One truncated fimbrin-GFP fusion was capable of labeling distinct filaments in stably transformed Arabidopsis roots. The filaments decorated by this construct were highly dynamic in growing root hairs and elongating root cells and were sensitive to actin disrupting drugs. Therefore, the fimbrin-GFP reporters we describe in this study provide additional tools for studying the actin cytoskeleton during root cell development. Moreover, the localization of AtFim1 GFP offers insights into the regulation of actin organization in developing roots by this class of actin cross-linking proteins. PMID- 15362113 TI - Visualization of a highly organized intranuclear network of filaments in living mammalian cells. AB - For 30 years, the mammalian cell nucleus has been hypothesized to contain a filamentous framework, the nuclear matrix or karyoskeleton, which regulates nuclear structure and function. However, such an organized network of filaments has never been observed in living cells. Here we show that human Cdc14B phosphatase in living cells tightly associates with long filaments that begin at the nucleolar periphery and extend to the nuclear envelope, frequently making close connections with nuclear pore complexes. We demonstrate that Cdc14B contains a bipartite signal that directs it to the intranuclear filaments, and we also detect a small amount of Cdc14B on interphase and mitotic centrosomes. Furthermore, we show that Cdc14B is critical for the maintenance of proper nuclear structure together with polo-like kinase Plk1. This work provides the first direct evidence for the existence of an intranuclear filamentous framework in living mammalian cells and implicates Cdc14B in the control of mammalian nuclear architecture. PMID- 15362114 TI - Destruction of maternal centrioles during fertilization of the brown alga, Scytosiphon lomentaria (Scytosiphonales, Phaeophyceae). AB - In brown algal fertilization, a pair of centrioles is derived from the male gamete, irrespective of the sexual reproduction pattern, i.e., isogamy, anisogamy, or oogamy. In this study, the manner in which the maternal centriole structure is destroyed in early zygotes of the isogamous brown alga Scytosiphon lomentaria was examined by electron microscopy. At fertilization, the zygote had two pairs of centrioles (flagellar basal bodies) derived from motile male and female gametes, and there was no morphological difference between the two pairs. The flagellar basal plate and the axonemal microtubules were still connected with the distal end of centrioles. Ultrastructural observations showed that the integrity of maternal-derived centrioles began to degenerate even in the 1-h-old zygote. At that time, the cylinder of triplet microtubules of the maternal centrioles became shorter from the distal end, and a section passing through the centrioles indicated that a part of the nine triplets of microtubules changed into doublet or singlet microtubules by degeneration of B and/or C tubules. In 2 h-old zygote, there was no trace of maternal centrioles ultrastructurally, and only the paternal centrioles remained. Further, reduction of centrin accompanying destruction of the maternal centrioles was examined in immunofluorescence microscopy. Centrin localized at the paternal and the maternal centrioles had the same fluorescence intensity in the early zygotes. At 4-6 h after fertilization, two spots indicating centrin localization showed different fluorescence intensity. Later, the weaker spot disappeared completely. These results showed that there is a difference in time between the destruction of the centriolar cylinders and the reduction of centrin molecules around them. PMID- 15362115 TI - Elusive nucleoskeleton visualized in live cells. PMID- 15362116 TI - Motility of a biflagellate sperm: waveform analysis and cyclic nucleotide activation. AB - The sperm of the freshwater clam Corbicula fluminea are unusual in that they have two flagella, both of which are capable of beating. When Corbicula sperm are removed from the gonad and placed into freshwater, most remain immotile. Video microscopy was used to assess signaling molecules capable of activating Corbicula sperm motility. Experiments using the cAMP analogs dbcAMP or 8-Br-cAMP show that elevating cAMP activates flagellar motility. Treatments with 8-Br-cGMP activated motility in similar numbers of sperm. Treatments with the selective cAMP dependent protein kinase (PKA) inhibitor H-89 block activation by 8-Br-cAMP but not by 8-Br-cGMP. Similar treatments with the cGMP-dependent protein kinase (PKG) inhibitor Rp-8-pCPT-cGMPS block activation by 8-Br-cGMP but not by 8-Br-cAMP. These results suggest that cAMP and cGMP each work through their specific kinase to activate flagellar motility. Analysis of spontaneously activated freely swimming sperm shows that the two flagella beat with different parameters. The A flagellum beats with a shorter wavelength and a higher frequency than the B flagellum. The observed differences in flagellar waveform indicate that the flagella are differentially controlled. PMID- 15362117 TI - Dynamic change in the distribution of alpha5beta1 integrin on isolated ventral membrane: effect of divalent cation species. AB - We investigated the spatial distribution of alpha5beta1 integrin in isolated ventral plasma membranes (VPMs) of human foreskin fibroblasts in order to elucidate how the interaction of integrin with cytoskeletal and extracellular matrix proteins might affect the integrin distribution. Cells were exposed to the jet of buffer to remove the apical surface and most of cellular organelles. After this treatment VPMs, which adhered to the glass surface, possessed the cellular structures such as fibronectin (FN) fibrils and actin stress fibers. The isolated VPMs thus prepared were employed without fixation to investigate the change in the integrin distribution. In isolated VPMs, alpha5beta1 integrin, labeled with Cy3-tagged anti-integrin antibody, was found to accumulate not only at the tips of stress fibers but also along FN fibrils extending from there. When divalent cations were removed with EDTA, the accumulated integrin was dispersed, and the original pattern of distribution was recovered upon restoration of divalent cations. Talin, an integrin-actin cytoskeleton linker protein, was found to accumulate only at the tips of stress fibers in isolated VPMs, but alpha5beta1 integrin did not exhibit strong accumulation there, indicating that talin played little role in integrin distribution in isolated VPMs. The amount of alpha actinin associated with stress fibers was found to drastically decrease in isolated VPMs, which was presumably related to the failure of localization of integrin at the tips of stress fibers. It was also shown that the association of stress fibers to isolated VPMs seemed to be independent of accumulation of integrin. PMID- 15362118 TI - Cellular localization of activated N-WASP using a conformation-sensitive antibody. AB - The main regulators of Arp2/3 activity appear to be N-WASP and the other members of the Scar/WAVE family of proteins. We show here that after EGF stimulation, N WASP is recruited to the nucleation zone of the dynamic leading edge compartment of carcinoma cells, with maximal recruitment of N-WASP within 1 min after EGF stimulation. The timing of N-WASP recruitment mirrors the timing of barbed-end formation at the leading edge. To determine the cellular activation of N-WASP after EGF stimulation, we made a conformation-sensitive antibody (CSA) against the CRIB domain of N-WASP that is predicted to recognize N-WASP in its open, active conformation, but not in its closed, inactive conformation. The ability of CSA to detect only active N-WASP was demonstrated by in vitro experiments using immunoprecipitation of active N-WASP from EGF-stimulated cells and Cdc42 activation of N-WASP activity. In cell staining experiments, N-WASP is maximally accessible to CSA 40 sec after EGF stimulation and this activated N-WASP is in the nucleation zone. These results indicate that active N-WASP is present at the leading edge of lamellipods, an unexpected finding given its reported involvement in filopod formation. This work establishes the feasibility of using antibodies directed against specific conformations or epitopes with changing accessibilities as a window on the status and localization of activity. PMID- 15362119 TI - Synthesis and antinociceptive activity of (5-chloro-2(3H)-benzothiazolon-3 yl)propanamide derivatives. AB - Ten new (5-chloro-2(3H)-benzothiazolon-3-yl)propanamide derivatives have been synthesized. The compounds were tested for antinociceptive activity by tail clip, tail flick, hot plate and writhing test by using aspirin and dipyrone as standards. Among these compounds, 1-[3-(5-chloro-2(3H)-benzothiazolon-3-yl) propanoyl]-4-(4-chlorophenyl)piperazine (11e) has been found to be significantly more active than the other compounds synthesized as well as the standards in all tests. PMID- 15362120 TI - Synthesis and molluscicidal activity of 5-oxo-5,6,7,8-tetrahydro-4H-chromene derivatives. AB - Furfurylidenemalononitriles and thienylidenemalononitriles were treated with 1,3 cyclohexanediones to afford 2-amino-4-hetaryl-5-oxo-5,6,7,8-tetrahydro-4H chromene-3-carbonitrile derivatives. The molluscicidal activity of these compounds was investigated. PMID- 15362121 TI - CDK1-inhibitory activity of paullones depends on electronic properties of 9 substituents. AB - Multiple linear regression analysis was employed in an effort to establish a quantitative structure-activity relationship model for the CDK1-inhibitory activity of a series of 9-substituted paullones. While the electronic properties of the 9-substituents proved to be of high relevance for CDK1 inhibition, both lipophilic and a steric parameters could not be included in a meaningful equation for the calculation of biological properties. The equation solely based on the electronic parameter was successfully used for the prediction of the CDK1 inhibitory activity of a small test set comprising novel paullones with sulfur containing 9-substituents. Among these new derivatives, 2-methoxy-9 methylsulfonylpaullone proved to be superior to the standard alsterpaullone with respect to CDK1 inhibition. PMID- 15362122 TI - Non-thiol farnesyltransferase inhibitors: utilization of the far aryl binding site by 5-cinnamoylaminobenzophenones. AB - We recently described two novel aryl binding sites of farnesyltransferase. In this study, the cinnamoyl residue was designed as an appropriate substituent for our benzophenone-based AAX-peptidomimetic compound capable of occupying the far aryl binding site. PMID- 15362123 TI - Modeling of the D2 dopamine receptor arylpiperazine binding site for 1-[2-[5-(1H benzimidazole-2-thione)]ethyl]-4-arylpiperazines. AB - Docking of several 1-[2-[5-(1H-benzimidazole-2-thione)]ethyl]-4- and 1-benzyl arylpiperazines to the D(2) dopamine receptor (DAR) was examined. The binding pocket of the D(2) DAR defined according to Teeter and DuRand [1] was extended using the Insight II software. It was found that (i) the interaction of the protonated N1 of the piperazine ring with Asp86, (ii) the hydrogen bond formation between the benzimidazole part of the ligand and Ser141, as well as Ser122, and (iii) the edge-to-face interactions of the aromatic ring or arylpiperazine part of the ligand with Phe178, Tyr216 and Trp182 of the receptor represent the mayor stabilizing forces. Besides, the hydrogen bond acceptor group in position 2 of the phenylpiperazine aromatic ring could form one more hydrogen bond with Trp182. Bulky substituents in position 4 are not tolerated, due to the unfavorable sterical interaction with Phe178. Substituents in positions 2 and 3 are sterically well tolerated. Electron-attractive groups (F, Cl, CF(3), and NO(2)) decreased, while electron donors (-OMe) and the second aromatic ring (naphthyl) increased the binding affinity, as compared to that of the parent compound 1. This can be explained by strong edge-to-face interactions of negative electrostatic surface potential (ESP) in the center of aromatic residues of the ligand with positive-ESP protons in the aromatic residues of the receptor. Thus, besides the salt bridges and hydrogen bonds, edge-to-face interactions significantly contribute to arylpiperazine ligands forming complexes with the D(2) DAR. PMID- 15362125 TI - PM3-compatible zinc parameters optimized for metalloenzyme active sites. AB - Recent studies have shown that semiempirical methods (e.g., PM3 and AM1) for zinc containing compounds are unreliable for modeling structures containing zinc ions with ligand environments similar to those observed in zinc metalloenzymes. To correct these deficiencies a reparameterization of zinc at the PM3 level was undertaken. In this effort we included frequency corrected B3LYP/6-311G* zinc metalloenzyme ligand environments along with previously utilized experimental data. Average errors for the heats of formation have been reduced from 46.9 kcal/mol (PM3) to 14.2 kcal/mol for this new parameter set, termed ZnB for "Zinc, Biological." In addition, the new parameter sets predict geometries for the Bacillus fragilis active site model and other zinc metalloenzyme mimics that are qualitatively in agreement with high-level ab initio results, something existing parameter sets failed to do. PMID- 15362126 TI - A dynamic lattice searching method for fast optimization of Lennard-Jones clusters. AB - A highly efficient unbiased global optimization method called dynamic lattice searching (DLS) was proposed. The method starts with a randomly generated local minimum, and finds better solution by a circulation of construction and searching of the dynamic lattice (DL) until the better solution approaches the best solution. The DL is constructed adaptively based on the starting local minimum by searching the possible location sites for an added atom, and the DL searching is implemented by iteratively moving the atom located at the occupied lattice site with the highest energy to the vacant lattice site with the lowest energy. Because the DL can greatly reduce the searching space and the number of the time consuming local minimization procedures, the proposed DLS method runs at a very high efficiency, especially for the clusters of larger size. The performance of the DLS is investigated in the optimization of Lennard-Jones (LJ) clusters up to 309 atoms, and the structure of the LJ(500) is also predicted. Furthermore, the idea of dynamic lattice can be easily adopted in the optimization of other molecular or atomic clusters. It may be a promising approach to be universally used for structural optimizations in the chemistry field. PMID- 15362127 TI - Solvation effects on alanine dipeptide: A MP2/cc-pVTZ//MP2/6-31G** study of (Phi, Psi) energy maps and conformers in the gas phase, ether, and water. AB - The effects of solvation on the conformations and energies of alanine dipeptide (AD) have been studied by ab initio calculations up to MP2/cc-pVTZ//MP2/6-31G**, utilizing the polarizable continuum model (PCM) to mimic solvation effects. The energy surfaces in the gas phase, ether, and water bear similar topological features carved by the steric hindrance, but the details differ significantly due to the solvent effects. The gas-phase energy map is qualitatively consistent with the Ramachandran plot showing seven energy minima. With respect to the gas-phase map, the significant changes of the aqueous map include (1) the expanded low energy regions, (2) the emergence of an energy barrier between C5-beta and alpha(R)-beta(2) regions, (3) a clearly pronounced alpha(R) minimum, a new beta conformer, and the disappearance of the gas-phase global minimum, and (4) the shift of the dominant region in LEII from the gas-phase C7(ax) region to the alpha(L) region. These changes bring the map in water to be much closer to the Ramachandran plot than the gas-phase map. The solvent effects on the geometries include the elongation of the exposed N-H and C=O bonds, the shortening of the buried HN--CO peptide bonds, and the enhanced planarity of the peptide bonds. The energy surface in ether has features similar to those both in the gas phase and in water. The free energy order computed in the gas phase and in ether is in good agreement with experimental studies that concluded that C5 and C7(eq) are the dominant species in both the gas phase and nonpolar solvents. The free energy order in water is consistent with the experimental observation that the dominant C7(eq) in the nonpolar solvent was largely replaced by P(II)-like (i.e., beta) and alpha(R) in the strong polar solvents. Based on calculations on AD + 4H(2)O and other AD-water clusters, we suggest that explicit water-AD interactions may distort C5 and beta (or alpha(R) and beta) to an intermediate conformation. Our analysis also shows that the PCM calculations at the MP2/cc-pVTZ//MP2/6-31G** level give good descriptions to the bulk solvent polarization effect. The results presented in this article should be of sufficient quality to characterize the peptide bonds in the gas phase and solvents. The energy surfaces may serve as the basis for developing of strategies enabling the inclusion of solvent polarization in the force field. PMID- 15362128 TI - Component-based integration of chemistry and optimization software. AB - Typical scientific software designs make rigid assumptions regarding programming language and data structures, frustrating software interoperability and scientific collaboration. Component-based software engineering is an emerging approach to managing the increasing complexity of scientific software. Component technology facilitates code interoperability and reuse. Through the adoption of methodology and tools developed by the Common Component Architecture Forum, we have developed a component architecture for molecular structure optimization. Using the NWChem and Massively Parallel Quantum Chemistry packages, we have produced chemistry components that provide capacity for energy and energy derivative evaluation. We have constructed geometry optimization applications by integrating the Toolkit for Advanced Optimization, Portable Extensible Toolkit for Scientific Computation, and Global Arrays packages, which provide optimization and linear algebra capabilities. We present a brief overview of the component development process and a description of abstract interfaces for chemical optimizations. The components conforming to these abstract interfaces allow the construction of applications using different chemistry and mathematics packages interchangeably. Initial numerical results for the component software demonstrate good performance, and highlight potential research enabled by this platform. PMID- 15362129 TI - Hybridized particle swarm algorithm for adaptive structure training of multilayer feed-forward neural network: QSAR studies of bioactivity of organic compounds. AB - The multilayer feed-forward ANN is an important modeling technique used in QSAR studying. The training of ANN is usually carried out only to optimize the weights of the neural network and without paying attention to the network topology. Some other strategies used to train ANN are, first, to discover an optimum structure of the network, and then to find weights for an already defined structure. These methods tend to converge to local optima, and may also lead to overfitting. In this article, a hybridized particle swarm optimization (PSO) approach was applied to the neural network structure training (HPSONN). The continuous version of PSO was used for the weight training of ANN, and the modified discrete PSO was applied to find appropriate the network architecture. The network structure and connectivity are trained simultaneously. The two versions of PSO can jointly search the global optimal ANN architecture and weights. A new objective function is formulated to determine the appropriate network architecture and optimum value of the weights. The proposed HPSONN algorithm was used to predict carcinogenic potency of aromatic amines and biological activity of a series of distamycin and distamycin-like derivatives. The results were compared to those obtained by PSO and GA training in which the network architecture was kept fixed. The comparison demonstrated that the HPSONN is a useful tool for training ANN, which converges quickly towards the optimal position, and can avoid overfitting in some extent. PMID- 15362130 TI - A new algorithm to handle finite nuclear mass effects in electronic calculations: the ISOTOPE program. AB - We report the development of a simple algorithm to modify quantum chemistry codes based on the LCAO procedure, to account for the isotope problem in electronic structure calculations. No extra computations are required compared to standard Born-Oppenheimer calculations. An upgrade of the Gamess package called ISOTOPE is presented, and its applicability is demonstrated in some examples. PMID- 15362131 TI - Ab initio and DFT studies on hydrolyses of phosphorus halides. AB - Hydrolyses of phosphorus halides, (RO)(2)POX where R = H or Me and X = F or Cl, in the gas phase and in the reaction field have been investigated theoretically with ab initio and the density functional theory (DFT). The free energy of activation in the reaction field was also estimated using the Onsager method with a correction of entropy change and basis set superposition error (BSSE). The reaction of (MeO)(2)POF proceeds through a path with bifunctional catalysis regardless of the medium, but the reaction of (MeO)(2)POCl proceeds through bifunctional and general base catalysis in the gas phase and in water, respectively. The estimated free energy barrier of 23 kcal/mol for the hydrolysis of (MeO)(2)POF is in good agreement with the experimental values of 24 kcal/mol, and relative barrier of 3 kcal/mol to the (MeO)(2)POCl is also in good agreement with the experimental values of 5 kcal/mol of diisopropyl phosphorus halides ((Pr(i)O)(2)POX, X = F and Cl). PMID- 15362132 TI - Efficient use of nonequilibrium measurement to estimate free energy differences for molecular systems. AB - A promising method for calculating free energy differences DeltaF is to generate nonequilibrium data via "fast-growth" simulations or by experiments--and then use Jarzynski's equality. However, a difficulty with using Jarzynski's equality is that DeltaF estimates converge very slowly and unreliably due to the nonlinear nature of the calculation--thus requiring large, costly data sets. The purpose of the work presented here is to determine the best estimate for DeltaF given a (finite) set of work values previously generated by simulation or experiment. Exploiting statistical properties of Jarzynski's equality, we present two fully automated analyses of nonequilibrium data from a toy model, and various simulated molecular systems. Both schemes remove at least several k(B)T of bias from DeltaF estimates, compared to direct application of Jarzynski's equality, for modest sized data sets (100 work values), in all tested systems. Results from one of the new methods suggest that good estimates of DeltaF can be obtained using 5-40-fold less data than was previously possible. Extending previous work, the new results exploit the systematic behavior of bias due to finite sample size. A key innovation is better use of the more statistically reliable information available from the raw data. PMID- 15362133 TI - The parameterization and validation of generalized born models using the pairwise descreening approximation. AB - Generalized Born Surface Area (GBSA) models for water using the Pairwise Descreening Approximation (PDA) have been parameterized by two different methods. The first method, similar to that used in previously reported parameterizations, optimizes all parameters against the experimental free energies of hydration of organic molecules. The second method optimizes the PDA parameters to compensate only for systematic errors of the PDA. The best models are compared to Poisson Boltzmann calculations and applied to the computation of potentials of mean force (PMFs) for the association of various molecules. PMFs present a more rigorous test of the ability of a solvation model to correctly reproduce the screening of intermolecular interactions by the solvent, than its accuracy at predicting free energies of hydration of small molecules. Models derived with the first method are sometimes shown to fail to compute accurate potentials of mean force because of large errors in the computation of Born radii, while no such difficulties are observed with the second method. Furthermore, accurate computation of the Born radii appears to be more important than good agreement with experimental free energies of solvation. We discuss the source of errors in the potentials of mean force and suggest means to reduce them. Our findings suggest that Generalized Born models that use the Pairwise Descreening Approximation and that are derived solely by unconstrained optimization of parameters against free energies of hydration should be applied to the modeling of intermolecular interactions with caution. PMID- 15362134 TI - The effect of basis set superposition error on the convergence of intermolecular interaction energies for deprotonated complexes. AB - The intermolecular interaction energies of the deprotonated hydrogen-bonded complexes F(-)(HF), F(-)(H(2)O), F(-)(NH(3)), Cl(-)(HF), SH(-)(HF), H(2)P(-)(HF), OH(-)(H(2)O), OH(-)(H(2)O)(2), OH(-)(NH(3)), Cl(-)(H(2)O), SH(-)(H(2)O), H(2)P( )(H(2)O), Cl(-)(NH(3)), SH(-)(NH(3)), H(2)P(-)(NH(3)), Cl(-)(HCl), Cl(-)(H(2)S), Cl(-)(PH(3)), SH(-)(H(2)S), SH(-)(PH(3)), and H(2)P(-)(PH(3)) were calculated with correlation consistent basis sets at the MP2, MP4, QCISD(T), and CCSD(T) levels. When the basis set is smaller, the counterpoise-uncorrected intermolecular interaction energies are closer to the complete basis set limit than the counterpoise-corrected intermolecular interaction energies. The counterpoise-uncorrected intermolecular interaction energies obtained at the MP2/aug-cc-pVDZ level of theory are close to the interaction energies obtained at the extrapolated complete basis set limit in most of the complexes. Also, we investigate the accuracy of the other levels. PMID- 15362135 TI - Kinetics and equilibrium of multicomponent adsorption on chiraly templated surfaces. AB - In this contribution we propose a simple model of adsorption of a binary (racemic) mixture on a chiraly templated surface. As an example, the adsorption of a liquid mixture of enantiomers on a chiral stationary phase (CSP) is considered. In particular, we study the effect of the lateral interactions in the adsorbed phase on the kinetic and equilibrium isotherms of the enantiomers. Additionally, we investigate the influence of the composition of the surface on the performance of the CSP in the presence of the lateral interactions. To that end, the adsorption of the mixture is modeled by using Monte Carlo simulations as well as by applying an analytical approach involving rate equations coupled with the Mean Field Approximation (MFA). The predictions of the theory are found to be in good agreement with the results of the simulations. PMID- 15362136 TI - Quantum-connectivity descriptors in modeling solubility of environmentally important organic compounds. AB - Quantum-connectivity indices are topographic descriptors combining quantum chemical and topological information. They are used to describe the water solubility of a noncongeneric data set of organic compounds. A QSPR model is obtained with two quantum-connectivity indices that accounts for more than 90% of the variance in the water solubility of these chemicals. This model is compared to other five QSPR models using constitutional, electrostatic, geometric, quantum chemical, and topological descriptors calculated by CODESSA. None of these models accounts for more than 85% of the variance in water solubility of the compounds in this data set. The QSPR model obtained with quantum-connectivity indices is also better than that generated from the general pool of 508 CODESSA indices. Models with up to five variables were explored and compared with the model obtained here. It is shown that quantum-connectivity indices contain more structural information than other classes of descriptors at least for describing the water solubility of these 53 chemicals. Structural interpretation of the QSPR model developed as well as the role of the quantum-connectivity indices included in it are also analyzed. PMID- 15362137 TI - Preprocessing of rotamers for protein design calculations. AB - We have developed a process that significantly reduces the number of rotamers in computational protein design calculations. This process, which we call Vegas, results in dramatic computational performance increases when used with algorithms based on the dead-end elimination (DEE) theorem. Vegas estimates the energy of each rotamer at each position by fixing each rotamer in turn and utilizing various search algorithms to optimize the remaining positions. Algorithms used for this context specific optimization can include Monte Carlo, self-consistent mean field, and the evaluation of an expression that generates a lower bound energy for the fixed rotamer. Rotamers with energies above a user-defined cutoff value are eliminated. We found that using Vegas to preprocess rotamers significantly reduced the calculation time of subsequent DEE-based algorithms while retaining the global minimum energy conformation. For a full boundary design of a 51 amino acid fragment of engrailed homeodomain, the total calculation time was reduced by 12-fold. PMID- 15362140 TI - Research for nursing practice: what do we tell practitioners about nursing interventions? PMID- 15362141 TI - Spouses of persons with dementia: their healthcare problems, utilization, and costs. AB - Spouses of persons with dementia (PWD) often experience poor health outcomes related to the experience of living with the afflicted spouse. Using the Anderson and Aday Healthcare Utilization Model, we conducted a retrospective review of an administrative database from a private healthcare insurer to compare health problems that precipitate utilization, patterns of utilization, and costs of care of spouses of PWD (n = 979) to those of spouses of persons without dementia (n = 979). Spouses of PWD were treated for more anxiety disorders (OR = 2.97; 95% CI = 1.63-5.44), falls (OR = 7.72; 95% CI = 2.73-21.84), rheumatologic diseases (OR = 2.5; 95% CI = 1.24-5.06), and diabetes with no complications (OR = 1.53; 95% CI = 1.06-2.22), but less pneumonia (OR =.55; 95%; CI =.35-.88) than comparison spouses. Spouses of PWD had a higher number of emergency room (ER) visits (p =.01). There were no differences in costs between the groups. The findings can be used to develop interventions for spouses of PWD. PMID- 15362142 TI - Helping at the bedside: spouses' preferences for helping critically ill patients. AB - Spouses of patients in intensive care units (ICU) need to be close and helpful to ill partners. According to adult attachment theory, emotional responses may be related to preferences for closeness and helpfulness, and according to control theory optimism also may influence spouses' emotional responses. Spouses' goals and helping behaviors were assessed in 88 spouses of ICU patients. Using a repeated-measures design, the relationships of closeness, helpfulness, and optimism to emotional outcomes were assessed. Preferences for closeness and helpfulness were strongly related, and together with optimism, predicted spouses' mood at some point of the illness trajectory. Spouses who were over-involved with partners' care requirements were at greater risk for emotional distress. Results suggest that closeness and helpfulness are integrated concepts, and that attachment dimensions of a relationship and optimism are useful for understanding spouses' emotional responses to critical illness. PMID- 15362143 TI - Efficacy of an in-home nursing intervention following short-stay breast cancer surgery. AB - This randomized controlled trial (n = 240) was designed to test the efficacy of a sub-acute home nursing intervention following short-stay surgery for breast cancer. Intervention participants received the in-home nursing protocol, whereas non-intervention participants received agency nursing care or no nursing care. Data, collected via questionnaire, telephone interview, and chart audit, included surgical recovery/self-care knowledge, functional status, anxiety, quality of life (QOL), and health service utilization. There were no significant group differences on postoperative functional status, anxiety, QOL, further surgeries, or complications. Intervention participants were more likely to receive instruction on surgical self-care (p 43 degrees C for TRPV1, >52 degrees C for TRPV2, > approximately 34-38 degrees C for TRPV3, > approximately 27-35 degrees C for TRPV4, < approximately 25-28 degrees C for TRPM8 and <17 degrees C for TRPA1), and are expressed in primary sensory neurons as well as other tissues. The involvement of TRPV1 in thermal nociception has been demonstrated by multiple methods, including the analysis of TRPV1-deficient mice. TRPV2, TRPM8, and TRPA1 are also very likely to be involved in thermal nociception, because their activation thresholds are within the noxious range of temperatures. PMID- 15362150 TI - Temperature perception and nociception. AB - The specificity theory of somesthesis holds that perceptions of warmth, cold, and pain are served by separate senses. Although no longer accepted in all its details, the theory's basic assumptions of anatomical and functional specificity have remained guiding principles in research on temperature perception and its relationship to pain. This article reviews the response characteristics of thermoreceptors, temperature-sensitive nociceptors, and their associated pathways in the context of old and new perceptual phenomena, most of which cannot be satisfactorily explained by the specificity theory. The evidence indicates that throughout most of the perceptual range, temperature sensitivity depends upon coactivation of, and interactions among, thermal and nociceptive pathways that are composed of both specific "labeled lines" and nonspecific, multimodal fibers. Adding to this complexity is evidence that tactile stimulation can influence the way in which thermal stimulation is perceived. It is argued that thermoreception is best defined as a functional subsystem of somesthesis that serves the very different and sometimes conflicting demands of thermoregulation, protection from thermal injury, and haptic perception. PMID- 15362151 TI - Mechanosensation and pain. AB - The ability of cells to detect and transduce mechanical stimuli impinging on them is a fundamental process that underlies normal cell growth, hearing, balance, touch, and pain. Surprisingly, little research has focused on mechanotransduction as it relates to the sensations of somatic touch and pain. In this article we will review data on the wealth of different mechanosensitive sensory neurons that innervate our main somatic sense organ the skin. The role of different types of mechanosensitive sensory neurons in pain under physiological and pathophysiological conditions (allodynia and hyperalgesia) will also be reviewed. Finally, recent work on the cellular and molecular mechanisms by which mechanoreceptive sensory neurons signal both innocuous and noxious sensation is evaluated in the context of pain. PMID- 15362152 TI - Understanding the signaling and transmission of visceral nociceptive events. AB - Visceral pain can be considered as part of the defense reactions of the body against harmful stimuli, particularly of those that impinge on the mucosal lining of hollow organs. It is a problem of considerable clinical relevance, and its neurobiological mechanisms differ from those of somatic nociceptive or neuropathic pain. Much progress had been made in recent years in the understanding of the functional properties of the visceral nociceptors that trigger pain states, their molecular mechanisms of activation and sensitization and on their central actions. Some molecular targets have been identified as key players in the activation and sensitization of visceral nociceptors, notably ASICs, TTX-resistant Na channels and the TRPV1 receptor. Some nonneural elements of visceral organs, such as the urothelium have been shown to play active roles in the transduction of visceral sensory events by mechanisms involving ATP release by the urothelial cells. Certain well-known neurotransmitters, such as the tachykinin family of neuropeptides, likely play an important role in the peripheral and central activation of visceral nociceptive afferents and in the generation of visceral hyperalgesia. This article reviews current evidence on the mechanisms of activation and sensitization of visceral nociceptive afferents and on their role in the triggering and maintenance of clinically relevant visceral pain states. PMID- 15362153 TI - Voltage-gated sodium channels and pain pathways. AB - Acute, inflammatory, and neuropathic pain can all be attenuated or abolished by local treatment with sodium channel blockers such as lidocaine. The peripheral input that drives pain perception thus depends on the presence of functional voltage-gated sodium channels. Remarkably, two voltage-gated sodium channel genes (Nav1.8 and Nav1.9) are expressed selectively in damage-sensing peripheral neurons, while a third channel (Nav1.7) is found predominantly in sensory and sympathetic neurons. An embryonic channel (Nav1.3) is also upregulated in damaged peripheral nerves and associated with increased electrical excitability in neuropathic pain states. A combination of antisense and knock-out studies support a specialized role for these sodium channels in pain pathways, and pharmacological studies with conotoxins suggest that isotype-specific antagonists should be feasible. Taken together, these data suggest that isotype-specific sodium channel blockers could be useful analgesics. PMID- 15362154 TI - Plasticity of pain signaling: role of neurotrophic factors exemplified by acid induced pain. AB - Acute noxious stimuli activate a specialized neuronal detection system that generates sensations of pain and, generally, adaptive behavioral responses. More persistent noxious stimuli notably those associated with some chronic injuries and disease states not only activate the pain-signaling system but also dramatically alter its properties so that weak stimuli produce pain. These hyperalgesic states arise from at least two distinct broad classes of mechanisms. These are peripheral and central sensitization associated with increased responsiveness of peripheral nociceptor terminals and dorsal horn neurons, respectively. Here we review the key features of these sensitized states and discuss the role of one neurotrophic factor, nerve growth factor, as a peripheral mediator of sensitization and of another factor, brain-derived neurotrophic factor, as a mediator of central sensitization. We use as a specific example the pain induced by acid stimuli. We review the neurobiology of such pain states, and discuss how acid stimuli both initiate sensitization and how the neuronal processing of acid stimuli is subject to sensitization. PMID- 15362155 TI - Posttranslational mechanisms of peripheral sensitization. AB - The sensation of pain can be dramatically altered in response to injury or disease. This sensitization can occur at the level of the primary sensory neuron, and can be mediated by multiple biochemical mechanisms, including, but not limited to, changes in gene transcription, changes in translation, stability, or subcellular localization of translated proteins, and posttranslational modifications. This review focuses on posttranslational modifications to ion channels expressed in primary sensory neurons that form the machinery driving peripheral sensitization and pain hypersensitivity. Studies published to date show strong evidence for modulation of ion channels involved in transduction and transmission of nociceptive inputs coincident with biophysical and behavioral sensitization. The roles of phosphorylation and oxidation/reduction reactions of voltage-dependent sodium, potassium, and calcium channels are discussed, as well as phosphorylation-mediated modulation of sensory transduction channels. PMID- 15362156 TI - Functional imaging of the human trigeminal system: opportunities for new insights into pain processing in health and disease. AB - Peripheral inflammation or nerve damage result in changes in nervous system function, and may be a source of chronic pain. A number of animal studies have indicated that central neural plasticity, including sensitization of neurons within the spinal cord and brain, is part of the response to nervous system insult, and can result in the appearance of altered sensation, including pain. It cannot be assumed, however, that data obtained from animal models unambiguously reflects CNS changes that occur in humans. Currently, the only noninvasive approach to determining objective changes in neural processing and responsiveness within the CNS in humans is the use of functional imaging techniques. It is now possible to use functional magnetic resonance imaging (fMRI) to measure CNS activation in the trigeminal ganglion, spinal trigeminal nucleus, the thalamus, and the somatosensory cortex in healthy volunteers, in a surrogate model of hyperalgesia, and in patients with trigeminal pain. By offering a window into the temporal and functional changes that occur in the damaged nervous system in humans, fMRI can provide both insight into the mechanisms of normal and pathological pain and, potentially, an objective method for measuring altered sensation. These advances are likely to contribute greatly to the diagnosis and treatment of clinical pain conditions affecting the trigeminal system (e.g., neuropathic pain, migraine). PMID- 15362157 TI - Antinociceptive and nociceptive actions of opioids. AB - Although the opioids are the principal treatment options for moderate to severe pain, their use is also associated with the development of tolerance, defined as the progressive need for higher doses to achieve a constant analgesic effect. The mechanisms which underlie this phenomenon remain unclear. Recent studies revealed that cholecystokinin (CCK) is upregulated in the rostral ventromedial medulla (RVM) during persistent opioid exposure. CCK is both antiopioid and pronociceptive, and activates descending pain facilitation mechanisms from the RVM enhancing nociceptive transmission at the spinal cord and promoting hyperalgesia. The neuroplastic changes elicited by opioid exposure reflect adaptive changes to promote increased pain transmission and consequent diminished antinociception (i.e., tolerance). PMID- 15362158 TI - The endogenous cannabinoid system and its role in nociceptive behavior. AB - The analgesic properties of exogenous cannabinoids have been recognized for many years and suggest a regulatory role for the endogenous cannabinoid ("endocannabinoid") system in mammalian nociceptive pathways. The endocannabinoid system includes: (1) at least two families of lipid signaling molecules, the N acyl ethanolamines (e.g., anandamide) and the monoacylglycerols (e.g., 2 arachidonoyl glycerol); (2) multiple enzymes involved in the biosynthesis and degradation of these lipids, including the integral membrane enzyme fatty acid amide hydrolase; and (3) two G-protein coupled receptors, CB1 and CB2, which are primarily localized to the nervous system and immune system, respectively. Here, we review recent genetic, behavioral, and pharmacological studies that have tested the function of the endocannabinoid system in pain sensation. Collectively, these investigations support a role for endocannabinoids in modulating behavioral responses to acute, inflammatory, and neuropathic pain stimuli. PMID- 15362159 TI - Invertebrate nociception: behaviors, neurons and molecules. AB - Genetic analysis of nociceptive behaviors in the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster has led to the discovery of conserved sensory transduction channels and signaling molecules. These are embedded in neurons and circuits that generate responses to noxious signals. This article reviews the neurons and molecular mechanisms that underlie invertebrate nociception. We begin with the neurobiology of invertebrate nociception, and then focus on molecules with conserved functions in vertebrate nociception and sensory biology. PMID- 15362160 TI - Epilepsy and the development of autism. Proceedings of a symposium. March 26-28, 2004, Sacramento, California, USA. PMID- 15362161 TI - Plasticity of nonneuronal brain tissue: roles in developmental disorders. AB - Neuronal and nonneuronal plasticity are both affected by environmental and experiential factors. Remodeling of existing neurons induced by such factors has been observed throughout the brain, and includes alterations in dendritic field dimensions, synaptogenesis, and synaptic morphology. The brain loci affected by these plastic neuronal changes are dependent on the type of experience and learning. Increased neurogenesis in the hippocampal dentate gyrus is a well documented response to environmental complexity ("enrichment") and learning. Exposure to challenging experiences and learning opportunities also alters existing glial cells (i.e., astrocytes and oligodendrocytes), and up-regulates gliogenesis, in the cerebral cortex and cerebellum. Such glial plasticity often parallels neuronal remodeling in both time and place, and this enhanced morphological synergism may be important for optimizing the functional interaction between glial cells and neurons. Aberrant structural plasticity of nonneuronal elements is a contributing factor, as is aberrant neuron plasticity, to neurological and developmental disorders such as epilepsy, autism, and mental retardation (i.e., fragile X syndrome). Some of these nonneuronal pathologies include abnormal cerebral and cerebellar white matter and myelin-related proteins in autism; abnormal myelin basic protein in fragile X syndrome (FXS); and abnormal astrocytes in autism, FXS, and epilepsy. A number of recent studies demonstrate the possibility of using environmental and experiential intervention to reduce or ameliorate some of the neuronal and nonneuronal abnormalities, as well as behavioral deficits, present in these neurological and developmental disorders. PMID- 15362162 TI - Environmental complexity and central nervous system development and function. AB - Environmental restriction or deprivation early in development can induce social, cognitive, affective, and motor abnormalities similar to those associated with autism. Conversely, rearing animals in larger, more complex environments results in enhanced brain structure and function, including increased brain weight, dendritic branching, neurogenesis, gene expression, and improved learning and memory. Moreover, in animal models of CNS insult (e.g., gene deletion), a more complex environment has attenuated or prevented the sequelae of the insult. Of relevance is the prevention of seizures and attenuation of their neuropathological sequelae as a consequence of exposure to a more complex environment. Relatively little attention, however, has been given to the issue of sensitive periods associated with such effects, the relative importance of social versus inanimate stimulation, or the unique contribution of exercise. Our studies have examined the effects of environmental complexity on the development of the restricted, repetitive behavior commonly observed in individuals with autism. In this model, a more complex environment substantially attenuates the development of the spontaneous and persistent stereotypies observed in deer mice reared in standard laboratory cages. Our findings support a sensitive period for such effects and suggest that early enrichment may have persistent neuroprotective effects after the animal is returned to a standard cage environment. Attenuation or prevention of repetitive behavior by environmental complexity was associated with increased neuronal metabolic activity, increased dendritic spine density, and elevated neurotrophin (BDNF) levels in brain regions that are part of cortical-basal ganglia circuitry. These effects were not observed in limbic areas such as the hippocampus. PMID- 15362163 TI - The effects of seizures on the connectivity and circuitry of the developing brain. AB - Recurring seizures in infants and children are often associated with cognitive deficits, but the reason for the learning difficulties is unclear. Recent studies in several animal models suggest that seizures themselves may contribute in important ways to these deficits. Other studies in animals have shown that recurring seizures result in dendritic spine loss. This change, coupled with a down-regulation in NMDA receptor subunit expression, suggests that repetitive seizures may interrupt the normal development of glutamatergic synaptic transmission. We hypothesize that homeostatic, neuroprotective processes are induced by recurring early-life seizures. These processes, by diminishing glutamatergic synaptic transmission, are aimed at preventing the continuation of seizures. However, by preventing the normal development of glutamatergic synapses, and particularly NMDA receptor-mediated synaptic transmission, such homeostatic processes also reduce synaptic plasticity and diminish the ability of neuronal circuits to learn and store memories. PMID- 15362164 TI - Effects of early seizures on later behavior and epileptogenicity. AB - Both clinical and laboratory studies demonstrate that seizures early in life can result in permanent behavioral abnormalities and enhance epileptogenicity. Understanding the critical periods of vulnerability of the developing nervous system to seizure-induced changes may provide insights into parallel or divergent processes in the development of autism. In experimental rodent models, the consequences of seizures are dependent on age, etiology, seizure duration, and frequency. Recurring seizures in immature rats result in long-term adverse effects on learning and memory. These behavioral changes are paralleled by changes in brain connectivity, changes in excitatory neurotransmitter receptor distribution, and decreased neurogenesis. These changes occur in the absence of cell loss. Although impaired cognitive function and brain changes have been well documented following early-onset seizures, the mechanisms of seizure-induced dysfunction remain unclear. PMID- 15362165 TI - Brain development in autism: early overgrowth followed by premature arrest of growth. AB - Due to the relatively late age of clinical diagnosis of autism, the early brain pathology of children with autism has remained largely unstudied. The increased use of retrospective measures such as head circumference, along with a surge of MRI studies of toddlers with autism, have opened a whole new area of research and discovery. Recent studies have now shown that abnormal brain overgrowth occurs during the first 2 years of life in children with autism. By 2-4 years of age, the most deviant overgrowth is in cerebral, cerebellar, and limbic structures that underlie higher-order cognitive, social, emotional, and language functions. Excessive growth is followed by abnormally slow or arrested growth. Deviant brain growth in autism occurs at the very time when the formation of cerebral circuitry is at its most exuberant and vulnerable stage, and it may signal disruption of this process of circuit formation. The resulting aberrant connectivity and dysfunction may lead to the development of autistic behaviors. To discover the causes, neural substrates, early-warning signs and effective treatments of autism, future research should focus on elucidating the neurobiological defects that underlie brain growth abnormalities in autism that appear during these critical first years of life. PMID- 15362166 TI - Serotonin in autism and pediatric epilepsies. AB - Serotonergic abnormalities have been reported in both autism and epilepsy. This association may provide insights into underlying mechanisms of these disorders because serotonin plays an important neurotrophic role during brain development- and there is evidence for abnormal cortical development in both autism and some forms of epilepsy. This review explores the hypothesis that an early disturbance in the serotonin system affects cortical development and the development of thalamocortical innervation, and is a potential mechanism, common to autism and pediatric epilepsies associated with cortical dysplasia. An argument is made that cortical malformation leads to abnormalities of thalamocortical connectivity, and that serotonin plays a critical role in this process. Finally, a role for altered metabolism of the serotonin precursur, tryptophan, in both epilepsy and autism is discussed. PMID- 15362167 TI - Sleep and arousal mechanisms in experimental epilepsy: epileptic components of NREM and antiepileptic components of REM sleep. AB - Neural generators related to different sleep components have different effects on seizure discharge. These sleep-related systems can provoke seizure discharge propagation during nonrapid eye movement (NREM) sleep and can suppress propagation during REM sleep. Experimental manipulations of discrete physiological components were conducted in feline epilepsy models, mostly in the systemic penicillin epilepsy model of primary generalized epilepsy and the amygdala kindling model of the localization-related seizure disorder, temporal lobe epilepsy. The sleep-wake state distribution of seizures was quantified before and after discrete lesions, systemic and localized drug administration, and/or photic stimulation, as well as in relation to microdialysis of norepinephrine. We found that (1) neural generators of synchronous EEG oscillations--including tonic background slow waves and phasic "arousal" events (sleep EEG transients such as sleep spindles and k-complexes)--combine to promote electrographic seizure propagation during NREM and drowsiness, and antigravity muscle tone permits seizure-related movement; (2) neural generators of asynchronous neuronal discharge patterns reduce electrographic seizures during alert waking and REM sleep, and skeletal motor paralysis blocks seizure-related movement during REM; (3) there are a number of similarities between amygdala kindled kittens and children with Landau-Kleffner Syndrome (LKS) that suggest a link among seizures, sleep disorders, and behavioral abnormalities/regression. PMID- 15362168 TI - Sleep disorders, epilepsy, and autism. AB - The purpose of this review article is to describe the clinical data linking autism with sleep and epilepsy and to discuss the impact of treating sleep disorders in children with autism either with or without coexisting epileptic seizures. Studies are presented to support the view that sleep is abnormal in individuals with autistic spectrum disorders. Epilepsy and sleep have reciprocal relationships, with sleep facilitating seizures and seizures adversely affecting sleep architecture. The hypothesis put forth is that identifying and treating sleep disorders, which are potentially caused by or contributed to by autism, may impact favorably on seizure control and on daytime behavior. The article concludes with some practical suggestions for the evaluation and treatment of sleep disorders in this population of children with autism. PMID- 15362169 TI - Neuroepileptic correlates of autistic symptomatology in tuberous sclerosis. AB - Tuberous sclerosis is a genetic condition that is strongly associated with the development of an autism spectrum disorder. However, there is marked variability in expression, and only a subset of children with tuberous sclerosis develop autism spectrum disorder. Clarification of the mechanisms that underlie the association and variability in expression will potentially throw light on the biological processes involved in the etiology of idiopathic forms of autism spectrum disorder. Current evidence indicates that the likelihood of a child with tuberous sclerosis developing an autism spectrum disorder is greater if the child has a mutation in the TSC2 gene, although autism can and does develop in children with TSC1 mutations. The likelihood is also greater if the child has early-onset infantile spasms that are difficult to control, especially if there is an epileptiform focus in the temporal lobes. The emerging evidence is consistent with the notion that early onset electrophysiological disturbances within the temporal lobes (and perhaps other locations) has a deleterious effect on the development and establishment of key social cognitive representations concerned with processing social information, perhaps especially from faces. However, alternative mechanisms to account for the findings cannot yet be ruled out. Future research will have to employ prospective longitudinal designs and treatment trials to clarify the processes involved. PMID- 15362170 TI - Epileptic encephalopathies and their relationship to developmental disorders: do spikes cause autism? AB - Epileptic encephalopathies are progressive clinical and electroencephalographic syndromes where deterioration is thought to be caused by frequent seizures and abundant EEG epileptiform activity. Seizures occur in approximately 10-15% of children with pervasive developmental disorders (PDD) and 8-10% have epileptiform EEG abnormalities without seizures. Thirty percent of children with PDD have regression of social behavior and language at 2-3 years of age. Some authors speculate that the regression is caused by epileptiform activity even in the absence of overt clinical seizures ("autism with epileptic regression") and suggest that elimination of the epileptiform activity, either medically or surgically, should lead to improvement in behavior. This review examines the data showing that interictal epileptiform discharges are associated with transient clinical dysfunction and discusses the implications of these observations for autistic behavioral abnormalities. The results of resective surgery, vagal nerve stimulation, and multiple subpial transaction on children with autism and epileptiform EEG abnormalities are also discussed. I conclude that there is no evidence that interictal discharges per se cause (or contribute to) the complex behavioral phenotype of autism. There is no justification to support the use of anticonvulsant medication or surgery in children with PDD without seizures; that is, there is no evidence that treatment to eliminate EEG spikes will have a therapeutic effect on the behavioral abnormalities of PDD and autism. PMID- 15362171 TI - AEDs and psychotropic drugs in children with autism and epilepsy. AB - The efficacy of antiepileptic drugs (AEDs) and psychotropic medications in children with autism is limited to the treatment of seizures or to specific behaviors such as irritability, impulsivity, hyperactivity, repetitive behaviors, or aggression. The reliability and value of the available data--to determine the efficacy of these medications in autism--are limited by lack of controlled clinical trials, the small number of subjects, the heterogeneity of the population studied, and the brief duration of most drug trials. Indeed, few controlled clinical trials using AEDs in autism, with or without seizures, have been conducted. Because some AEDs also have a positive effect on mood, the benefits that children with autism sometimes obtain from these medications may not be due to the treatment of the abnormal electrical activity or the seizures per se but to an effect on common neuronal systems responsible for both behavior and epilepsy. The relationship between epilepsy and autism, and specifically the effects that abnormal electrical activity may have on the developing brain, may provide some valuable insights into the type of studies that are needed to help us understand the pathophysiology of autism. PMID- 15362172 TI - Developmental regression in autism spectrum disorders. AB - The occurrence of developmental regression in autism is one of the more puzzling features of this disorder. Although several studies have documented the validity of parental reports of regression using home videos, accumulating data suggest that most children who demonstrate regression also demonstrated previous, subtle, developmental differences. Counter to clinical intuition, the earlier development of social, language, and attachment behaviors followed by regression does not seem to support later recovery of skills or better developmental outcomes compared to children who never had speech or typical social responsivity. In fact, this regressive group may have somewhat greater developmental impairment than the nonregressive group, though the two groups do not appear to present different behavioral phenotypes. Although autism is not the only condition in which regression occurs, it appears to be the most frequent condition. Other disorders that demonstrate an early regression with no known etiology include total blindness from birth and childhood disintegrative disorder, both of which demonstrate behavioral relations to autism. In addition, two biological conditions with known etiologies also involve regression with some behaviors resembling autism behavioral phenotype: Rett syndrome (a genetic disorder; see Glaze, this issue) and Landau-Kleffner syndrome (see McVicar and Shinnar, this issue), which involves a seizure disorder. PMID- 15362173 TI - Landau-Kleffner syndrome, electrical status epilepticus in slow wave sleep, and language regression in children. AB - The Landau-Kleffner syndrome (LKS) and electrical status epilepticus in slow wave sleep (ESES) are rare childhood-onset epileptic encephalopathies in which loss of language skills occurs in the context of an epileptiform EEG activated in sleep. Although in LKS the loss of function is limited to language, in ESES there is a wider spectrum of cognitive impairment. The two syndromes are distinct but have some overlap. The relationship between the epileptiform EEG abnormalities and the loss of cognitive function remains controversial, even in LKS which is the most widely accepted as an acquired epileptic aphasia. Language regression also occurs in younger children, frequently in the context of a more global autistic regression. Many of these children have epileptiform EEGs. The term autistic regression with epileptiform EEG has been proposed for these children. Whether these children are part of an extended LKS spectrum is very controversial, because there are differences in age of onset, clinical phenotype, and EEG findings. An understanding of the available data on clinical characteristics, EEG findings, pathology, prognosis, and treatment of these syndromes is essential for further progress in this area. PMID- 15362174 TI - Lennox-Gastaut syndrome: potential mechanisms of cognitive regression. AB - Lennox-Gastaut (L-G) syndrome is an intractable generalized epilepsy of childhood onset, associated with spike waves at a slow rate and paroxysmal fast activity. These epileptiform discharge patterns are thought to reflect excessive neocortical excitability and arise from neuronal and synaptic features peculiar to the immature central nervous system. The epileptic processes associated with L G syndrome may lead to enduring patterns of abnormal activity and connectivity. These abnormal patterns compete with normal developmental mechanisms and may result in subsequent impairment and/or regression of cognition. Recurring or prolonged seizures themselves may also damage the brain. We hypothesize that the presence of slow spike waves diverts the brain from normal developmental processes toward seizure-preventing mechanisms. Adding to this burden, antiepileptic medications, sleep disruption, and social isolation all retard cognitive development and the learning process at a crucial time of brain maturation. PMID- 15362175 TI - Rett syndrome: of girls and mice--lessons for regression in autism. AB - Rett syndrome (RTT) is a neurodevelopmental disorder occurring almost exclusively in females. Regression is a defining feature of RTT. During the regression stage, RTT girls display many autistic features, such as loss of communication and social skills, poor eye contact, and lack of interest, and initially may be given the diagnosis of autism. The discovery of the genetic cause of RTT, mutations in the MECP2 gene, a transcriptional repressor, has promoted the early diagnosis of RTT and development of mouse models. The phenotype of one mouse model includes features such as regression and abnormal behavioral and social interactions. The timing of the period of regression in RTT--during ages 1 to 2 years--parallels the period of intense synaptic development. The effects of the MECP2 mutation also increases concomitantly with peak synaptogenesis. Neuropathological findings in Rett include the selective reduction of dendritric spines in the pyramidal cells of RTT brains; this feature has also been reported in autism. Studies have observed that MECP influences the expression of brain-derived neurotrophic factor and thus may influence synaptic plasticity. Abnormalities in synapse maintenance and modulation may contribute to regression in RTT and autism. Studies of the clinical aspects of the regression period and of the mouse model may be useful in understanding the pathophysiology of RTT and other neurodevelopmental disorders such as autism. A recent study observed abnormal expression of MeCP2 in RTT and other neurodevelopmental disorders such as autism. Although the genetic background and certain clinical features differ in RTT and autism, a similar mechanism involving MeCP2 regulation and expression may contribute to regression. PMID- 15362176 TI - Alcohol and marijuana use among college students: economic complements or substitutes? AB - Previous research has shown that the recent tightening of college alcohol policies has been effective at reducing college students' drinking. Over the period in which these stricter alcohol policies have been put in place, marijuana use among college students has increased. This raises the question of whether current policies aimed at reducing alcohol consumption are inadvertently encouraging marijuana use. This paper begins to address this question by investigating the relationship between the demands for alcohol and marijuana for college students using data from the 1993, 1997 and 1999 waves of the Harvard School of Public Health's College Alcohol Study (CAS). We find that alcohol and marijuana are economic complements and that policies that increase the full price of alcohol decrease participation in marijuana use. PMID- 15362177 TI - Health insurance and treatment seeking behaviour: evidence from a low-income country. AB - This paper analyses the effect of being insured under the voluntary component of Vietnamese Health Insurance, on patterns of treatment seeking behaviour. A multinomial logit model is estimated using household survey data from three provinces in Vietnam. Decisions regarding both the type of provider sought and type of care received are analysed. Insurance status is treated as both exogenous and endogenous to account for potential selection bias. The results indicate that, overall, insured patients are more likely to use outpatient facilities, and public providers, an effect that is particularly strong at lower income levels. PMID- 15362178 TI - Money for health: the equivalent variation of cardiovascular diseases. AB - This paper introduces a new method to calculate the extent to which individuals are willing to trade money for improvements in their health status. An individual welfare function of income (WFI) is applied to calculate the equivalent income variation of health impairments. We believe that this approach avoids various drawbacks of alternative willingness-to-pay methods. The WFI is used to calculate the equivalent variation of cardiovascular diseases. It is found that for a 25 year old male the equivalent variation of a heart disease ranges from 114,000 euro to 380,000 euro depending on the welfare level. This is about 10,000 euro - 30,000 euro for an additional life year. The equivalent variation declines with age and is about the same for men and women. The estimates further vary by discount rate chosen. The estimates of the equivalent variation are generally higher than the money spent on most heart-related medical interventions per QALY. The cost-benefit analysis shows that for most interventions the value of the health benefits exceeds the costs. Heart transplants seem to be too costly and only beneficial if patients are young. PMID- 15362179 TI - A comparison of the EQ-5D and SF-6D across seven patient groups. AB - As the number of preference-based instruments grows, it becomes increasingly important to compare different preference-based measures of health in order to inform an important debate on the choice of instrument. This paper presents a comparison of two of them, the EQ-5D and the SF-6D (recently developed from the SF-36) across seven patient/population groups (chronic obstructive airways disease, osteoarthritis, irritable bowel syndrome, lower back pain, leg ulcers, post menopausal women and elderly). The mean SF-6D index value was found to exceed the EQ-5D by 0.045 and the intraclass correlation coefficient between them was 0.51. Whilst this convergence lends some support for the validity of these measures, the modest difference at the aggregate level masks more significant differences in agreement across the patient groups and over severity of illness, with the SF-6D having a smaller range and lower variance in values. There is evidence for floor effects in the SF-6D and ceiling effects in the EQ-5D. These discrepancies arise from differences in their health state classifications and the methods used to value them. Further research is required to fully understand the respective roles of the descriptive systems and the valuation methods and to examine the implications for estimates of the impact of health care interventions. PMID- 15362180 TI - The wage effects of obesity: a longitudinal study. AB - We use National Longitudinal Survey of Youth (NLSY) data to examine the effects of obesity on wages by gender. Sample means indicate that both men and women experience a persistent obesity wage penalty over the first two decades of their careers. We then control for a standard set of socioeconomic and familial variables but find that standard covariates do not explain why obese workers experience persistent wage penalties. This suggests that other variables -- including job discrimination, health-related factors and/or obese workers' behavior patterns -- may be the channels through which obesity adversely affects wages. The study closes with a discussion of the public policy implications suggested by these findings. PMID- 15362181 TI - Deriving welfare measures from discrete choice experiments: inconsistency between current methods and random utility and welfare theory. AB - Discrete choice experiments (DCEs) are being used increasingly in health economics to elicit preferences for products and programs. The results of such experiments have been used to calculate measures of welfare or more specifically, respondents' 'willingness to pay' (WTP) for products and programs and their 'marginal willingness to pay' (MWTP) for the attributes that make up such products and programs. In this note we show that the methods currently used to derive measures of welfare from DCEs in the health economics literature are not consistent with random utility theory (RUT), or with microeconomic welfare theory more generally. The inconsistency with welfare theory is an important limitation on the use of such WTP estimates in cost-benefit analyses. We describe an alternative method of deriving measures of welfare (compensating variation) from DCEs that is consistent with RUT and is derived using welfare theory. We demonstrate its use in an empirical application to derive the WTP for asthma medication and compare it to the results elicited from the method currently used in the health economics literature. PMID- 15362182 TI - Deriving welfare measures in discrete choice experiments: a comment to Lancsar and Savage (1). AB - Lancsar and Savage argue that current methods of deriving welfare estimates, using discrete choice experiments, are inconsistent with random utility and welfare theory. In this paper I show that this not the case. The general formula proposed by Small and Rosen for estimating welfare, which Lancsar and Savage claim should be used, reduces to the method used by health economists for state of the world models. The important question then becomes when are state of the world models, as opposed to multiple alternative models, appropriate? PMID- 15362183 TI - Deriving welfare measures in discrete choice experiments: a comment to Lancsar and Savage (2). PMID- 15362185 TI - Radio frequency heating: a new potential means of post-harvest pest control in nuts and dry products. AB - The multi-billion dollar US tree nut industries rely heavily on methyl bromide fumigation for postharvest insect control and are facing a major challenge with the mandated cessation by 2005 of its use for most applications. There is an urgent need to develop effective and economically viable alternative treatments to replace current phytosanitary and quarantine practices in order to maintain the competitiveness of US agriculture in domestic and international markets. With the reliable heating block system, the thermal death kinetics for fifth-instar codling moth, Indianmeal moth, and navel orangeworm were determined at a heating rate of 18 degrees C/min. A practical process protocol was developed to control the most heat resistant insect pest, fifth-instar navel orangeworm, in in-shell walnuts using a 27 MHz pilot scale radio frequency (RF) system. RF heating to 55 degrees C and holding in hot air for at least 5 min resulted in 100% mortality of the fifth-instar navel orangeworm. Rancidity, sensory qualities and shell characteristics were not affected by the treatments. If this method can be economically integrated into the handling process, it should have excellent potential as a disinfestation method for in-shell walnuts. PMID- 15362186 TI - A novel reduction of diketones with i-RMgBr catalyzed by Cp2TiCl2 and deoxygenation of sulfoxides by Cp2TiCl2/Al system. AB - Alpha-diketones and beta-diketones react with Grignard reagents in the presence of a catalytic amount of Cp2TiCl2 to yield alpha-ketols and corresponding ketones respectively. Sulfoxides can be deoxygenated by Cp2TiCl2/Al system. The possible mechanisms are also discussed. PMID- 15362187 TI - Discovery of ultrahigh-T spinel-garnet granulite with pure CO2 fluid inclusions from the Altay orogenic belt, NW China. AB - We first report discovery of the spinel-garnet-orthopyroxene granulite with pure CO2 fluid inclusions from the Fuyun region of the late Paleozoic Altay orogenic belt in Central Asia, NW China. The rock is characterized by an assemblage of garnet, orthopyroxene, spinel, cordierite, biotite, plagioclase and quartz. Symplectites of orthopyroxene and spinel, and orthopyroxene and cordierite indicate decompression under UHT conditions. Mineral chemistry shows that the orthopyroxenes have high XMg and Al2O3 contents (up to 9.23 wt%). Biotites are enriched in TiO2 and XMg and are stable under granulite facies conditions. The garnet and quartz from the rock carry monophase fluid inclusions which show peak melting temperatures of around -56.7 degrees C, indicating a pure CO2 species being presented during the ultrahigh-T metamorphism in the Altay orogenic belt. The inclusions homogenize into a liquid phase at temperatures around 15.3-23.8 degrees C translating into CO2 densities of the order of 0.86-0.88 g/cm3. Based on preliminary mineral paragenesis, reaction textures and petrogenetic grid considerations, we infer that the rock was subjected to UHT conditions. The CO2 rich fluids were trapped during exhumation along a clockwise P-T path following isothermal decompression under UHT conditions. PMID- 15362188 TI - Microbial control of diamondback moth, Plutella xylostella L. (Lepidoptera: Yponomeutidae) using bacteria (Xenorhabdus nematophila) and its metabolites from the entomopathogenic nematode Steinernema carpocapsae. AB - Cells and cell-free solutions of the culture filtrate of the bacterial symbiont, Xenorhabdus nematophila taken from the entomopathogenic nematode Steinernema carpocapsae in aqueous broth suspensions were lethal to larvae of the diamondback moth Plutella xylostella. Their application on leaves of Chinese cabbage indicated that the cells can penetrate into the insects in the absence of the nematode vector. Cell-free solutions containing metabolites were also proved as effective as bacterial cells suspension. The application of aqueous suspensions of cells of X. nematophila or solutions containing its toxic metabolites to the leaves represents a possible new strategy for controlling insect pests on foliage. PMID- 15362189 TI - Species-diversified plant cover enhances orchard ecosystem resistance to climatic stress and soil erosion in subtropical hillside. AB - Naturally occurring plants in agroecosystem evidently play an important role in ecosystem stability. Field studies on the ecological effects of native plants conserved in orchard and their resistance to adverse climatic stress, and soil erosion were conducted from 1998 to 2001 in a newly developed Changshan-huyou (Citrus changshan-huyou Y.B. Chang) orchard. The experimental area covered 150 ha in typical red soil hilly region in southeastern China. The experimental design was a randomized complete block with six combinations of twelve plant species with four replications. All species used were native in the orchard. Plots were 15 x 8 m2 and separated by 2 m buffer strips. Precipitation, soil erosion in rainstorm days and aboveground biomass of plant community when rainstorm days ended, soil temperature and moisture under various plant covers during seasonal megathermal drought period, antiscourability of soil with different root density under various simulated rainfalls were measured. Plant cover significantly decreased the daily highest and mean soil temperature and its daily variation in hot-drought season, but there was no significant difference of the alleviation among various plant covers. Plant covers significantly increased the soil moisture in seasonal megathermal drought period. Better moisture maintenance and soil erosion reduction was found when the plant species numbers in cover plant communities increased from one to eight. Higher root density in plant communities with higher species richness increased significantly the antiscourability of the soil. It was suggested that conserving plant communities with diversified native species could produce the best positive ecological effects on citrus orchard ecosystem stability. PMID- 15362190 TI - Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L. AB - Novel bacterial blight (BB) resistance gene(s) for rice was (were) introduced into a cultivated japonica rice variety Oryza sativa (cv. 8411), via somatic hybridization using the wild rice Oryza meyeriana as the donor of the resistance gene(s). Twenty-nine progenies of somatically hybridized plants were obtained. Seven somatically hybridized plants and their parents were used for AFLP (amplified fragment length polymorphism) analysis using 8 primer pairs. Results confirmed that these plants were somatic hybrids containing the characteristic bands of both parents. The morphology of the regenerated rice showed characters of both O. sativa and O. meyeriana. Two somatic hybrids showed highest BB resistance and the other 8 plants showed moderate resistance. The new germplasms with highest resistance have been used in the rice breeding program for the improvement of bacterial blight resistance. PMID- 15362191 TI - Prediction and setup of phytoplankton statistical model of Qiandaohu Lake. AB - This research considers the mathematical relationship between concentration of Chla and seven environmental factors, i.e. Lake water temperature (T), Secci depth (SD), pH, DO, CODMn, Total Nitrogen (TN), Total Phosphorus (TP). Stepwise linear regression of 1997 to 1999 monitoring data at each sampling point of Qiandaohu Lake yielded the multivariate regression models presented in this paper. The concentration of Chla as simulation for the year 2000 by the regression model was similar to the observed value. The suggested mathematical relationship could be used to predict changes in the lakewater environment at any point in time. The results showed that SD, TP and pH were the most significant factors affecting Chla concentration. PMID- 15362192 TI - Ultrastructure of spermatozoa of Bullacta exarata (philippi) and its significance on reproductive evolution and physio-ecological adaptation. AB - The morphology and ultrastructure of Bullacta exarata spermatozoa observed by light and transmission electron microscopy are presented in this paper. The spermatozoa is composed of head with a simple acrosomal complex and an elongated nucleus, and tail with a midpiece, principal piece and an end piece. The midpiece consists of a mitochondrial ring, and the principal piece is composed of axoneme and lateral fin. The structure of B. exarata spermatozoa differs significantly from that of other gastropods, especially in the lateral fin and the principal piece, which was described scarcely before. A comparison is made between B. exarata and other gastropods, and its significance on reproductive evolution and physio-ecological adaptation is preliminarily discussed. PMID- 15362193 TI - Sap flow response of Eucaylyptus (Eucalyptus urophylla) to environmental stress in South China. AB - Sap flow and environmental conditions were monitored at two Eucalyptus (Eucalyptus urophylla S.T.Blake) plantations at Hetou and Jijia, located in Leizhou, Zhanjiang, Guangdong Province. It was found that daily sap flux density (SFD) of Eucalyptus was closely related to daily atmospheric vapor pressure deficit (VPD) (R2 = 0.76, P = 0.01 at Hetou and R2 = 0.7021, P = 0.01 at Jijia) at both sites. No significant relationship existed between daily SFD and mean daily air temperature at both sites. Daily SFD varied with wind speed Y = 17585X3 + 15147X2 - 1250.7X + 2278.4 (R2 = 0.68; P = 0.01) at Hetou and Y = 101.67X3 - 1.65X2 - 376.4X + 1914.8 (R2 = 0.40, P = 0.05) at Jijia, where Y was daily SFD, X was daily wind speed. Experimental observations yielded the following data: (1) the critical lower and upper daily VPD threshold were 0 and 2 kPa, within which daily SFD varied from 540+/-70 L/(m2.d) to 4739+/-115 L/(m2.d) at Hetou site, from 397+/-26 L/(m2.d) to 3414+/-191 L/(m2.d) at Jijia site; (2) Diurnal SFDs at Hetou site were much higher under low relative humidity (< 30%) and slightly lower under high relative humidity (> 80%) compared with those at the Jijia site; (3) The upper and lower threshold of daily and diurnal RAD for the optimal water use of E. urophylla plantations were 18+/-2.7 and 2+/-1 MJ/(m2.d), 669 and 0 J/(cm2.h) during the observation period. PMID- 15362194 TI - Protein folding pathology in domestic animals. AB - Fibrillar proteins form structural elements of cells and the extracellular matrix. Pathological lesions of fibrillar microanatomical structures, or secondary fibrillar changes in globular proteins are well known. A special group concerns histologically amorphous deposits, amyloid. The major characteristics of amyloid are: apple green birefringence after Congo red staining of histological sections, and non-branching 7-10 nm thick fibrils on electron microscopy revealing a high content of cross beta pleated sheets. About 25 different types of amyloid have been characterised. In animals, AA-amyloid is the most frequent type. Other types of amyloid in animals represent: AIAPP (in cats), AApoAI, AApoAII, localised AL-amyloid, amyloid in odontogenic or mammary tumors and amyloid in the brain. In old dogs Abeta and in sheep APrPsc-amyloid can be encountered. AA-amyloidosis is a systemic disorder with a precursor in blood, acute phase serum amyloid A (SAA). In chronic inflammatory processes AA-amyloid can be deposited. A rapid crystallization of SAA to amyloid fibrils on small beta sheeted fragments, the 'amyloid enhancing factor' (AEF), is known and the AEF has been shown to penetrate the enteric barrier. Amyloid fibrils can aggregate from various precursor proteins in vitro in particular at acidic pH and when proteolytic fragments are formed. Molecular chaperones influence this process. Tissue data point to amyloid fibrillogenesis in lysosomes and near cell surfaces. A comparison can be made of the fibrillogenesis in prion diseases and in enhanced AA-amyloidosis. In the reactive form, acute phase SAA is the supply of the precursor protein, whereas in the prion diseases, cell membrane proteins form a structural source. Abeta-amyloid in brain tissue of aged dogs showing signs of dementia forms a canine counterpart of senile dementia of the Alzheimer type (ccSDAT) in man. Misfolded proteins remain potential food hazards. Developments concerning prevention of amyloidogenesis and therapy of amyloid deposits are shortly commented. PMID- 15362195 TI - TGF-beta1 treated murine dendritic cells are maturation resistant and down regulate Toll-like receptor 4 expression. AB - OBJECTIVE: To explore the effects of transforming growth factor beta1 (TGF-beta1) on dendritic cells (DC). METHODS: Murine bone marrow cells were cultured with GM CSF and TGF-beta1 to develop TGF-beta1-treated DC (TGFbeta-DC). Then they were stimulated by lipopolysaccharide (LPS). Their phenotypes were assessed by flow cytometry (FCM). The allogeneic stimulating capacity of DC was measured by mixed lymphocyte reaction (MLR) using BrdU ELISA method and IL-12p70 protein was detected by ELISA. The expression of Toll-like receptor 4 (TLR4) was analyzed by semi quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and FCM. RESULTS: Compared to immature DC (imDC) cultured by GM-CSF alone, the TGFbeta-DC express lower CD80, CD86, I-Ab and CD40. The TGFbeta-DC were resistant to maturation with LPS. Maturation resistance was evident from a failure to up regulate co-stimulatory molecules (CMs), to stimulate larger T cells proliferation and to enhance secretion of IL-12p70. We also found that TGF-beta1 could down-regulate TLR4 expression on TGFbeta-DC. CONCLUSION: TGFbeta-DC are resistant to maturation stimulus (LPS) and might have some correlation with the down-modulation of TLR4 expression. PMID- 15362196 TI - Effects of vanadate on the activities of mice glucokinase and hexokinase. AB - This study aimed at acquiring knowledge on the hypoglycemic mechanisms of sodium metavanadate (SMV) showed that the liver glucokinase and muscle hexokinase activities increased rapidly after oral SMV was given, and that the blood glucose level was correlated closely with the activities of the two enzymes but not with the insulin level; which indicated that SMV could improve the altered glucose phosphorylation in diabetic mice independently of stimulating insulin secretion. This was probably one of the mechanisms of hypoglycemic effects of SMV. PMID- 15362197 TI - Identification of embryonic chromosomal abnormality using FISH-based preimplantation genetic diagnosis. AB - OBJECTIVE: Embryonic chromosomal abnormality is one of the main reasons for in vitro fertilization (IVF) failure. This study aimed at evaluating the value of Fluorescence in-situ Hybridization (FISH)-based Preimplantation Genetic Diagnosis (PGD) in screening for embryonic chromosomal abnormality to increase the successful rate of IVF. METHOD: Ten couples, four with high risk of chromosomal abnormality and six infertile couples, underwent FISH-based PGD during IVF procedure. At day 3, one or two blastomeres were aspirated from each embryo. Biopsied blastomeres were examined using FISH analysis to screen out embryos with chromosomal abnormalities. At day 4, embryos without detectable chromosomal abnormality were transferred to the mother bodies as in regular IVF. RESULTS: Among 54 embryos screened using FISH-based PGD, 30 embryos were detected to have chromosomal abnormalities. The 24 healthy embryos were implanted, resulting in four clinical pregnancies, two of which led to successful normal birth of two healthy babies; one to ongoing pregnancy during the writing of this article; and one to ectopic pregnancy. CONCLUSION: FISH-based PGD is an effective method for detecting embryonic chromosomal abnormality, which is one of the common causes of spontaneous miscarriages and chromosomally unbalanced offsprings. PMID- 15362198 TI - Lymphotactin enhances the in-vitro immune efficacy of dendritoma formed by dendritic cells and mouse hepatocellular carcinoma cells. AB - OBJECTIVE: To investigate the in-vitro antitumor immune responses of dendritoma formed by mouse hepatocellular carcinoma (HCC) cells and lymphotactin (Lptn) gene modified dendritic cells (DCs). METHOD: DCs prepared from mouse bone marrow were genetically modified by lymphotactin adenovirus, and fused with H22 cells by polyethylene glycol (PEG). RT-PCR and ELISA were employed to identify lymphotactin expression at mRNA and protein level. Cell phenotypes and fusion efficiency was detected by FACS. The stimulatory effect of DC on T cells was detected by mixed lymphocyte reaction. The cytotoxicity activity against H22 cells was assayed by LDH method. RESULTS: Lymphotactin could be efficiently expressed by DCLptn/H22 hybridoma. DCLptn/H22 cells could induce potent T cell proliferation effect and generate strong cytotoxic T lymphocyte (CTL) reaction against allogenic H22 cells. CONCLUSION: Lymphotactin genetic modification could enhance the in vitro immune activity of the dendritoma. PMID- 15362199 TI - Study of clinical features of amyloid angiopathy hemorrhage and hypertensive intracerebral hemorrhage. AB - OBJECTIVE: The purpose of this study was to differentiate between cerebral amyloid angiopathy (CAA) and hypertension (HTN) based on hemorrhage pattern interpretation. METHODS: From June 1994 to Oct., 2000, 83 patients admitted to our service with acute intracerebral hemorrhage (ICH) were investigated retrospectively; 41 patients with histologically proven diagnosis of cerebral amyloid angiography and 42 patients with clear history of hypertension were investigated. RESULTS: Patients with a CAA-related ICH were significantly older than patients with a HTN-related ICH (74.0 years vs 66.5 years, P < 0.05). There was a significantly higher number of hematomas > or = 30 ml in CAA (85.3%) when compared with HTN (59.5%). No basal ganglional hemorrhage was seen in CAA, but in 40.5% in HTN. In CAA-related ICH, subarachnoid hemorrhage (SAH) was seen in 26 patients (63.4%) compared to only 11 patients (26.2%) in HTN-related ICH. Intraventricular hemorrhage was seen in 24.4% in CAA, and in 26.2% in HTN. Typical features of CAA-related ICH included lobar distribution affecting mainly the lobar superficial areas, lobulated appearance, rupture into the subarachnoid space, and secondary IVH from the lobar hemorrhage. More specifically, multiplicity of hemorrhage, bilaterality, and repeated episodes also strongly suggest the diagnosis of CAA. Multiple hemorrhages, defined as 2 or more separate hematomas in multiple lobes, accounted for 17.1% in CAA-related ICH. CONCLUSION: There are certain features in CAA on CT and MRI and in clinical settings. To some extent, these features may contribute to distinguishing CAA from HTN related ICH. PMID- 15362200 TI - The dimensional accuracy of preparation of femoral cavity in cementless total hip arthroplasty. AB - OBJECTIVE: To observe the accuracy of femoral preparation and the position of the cementless prosthesis in femoral cavity, and to compare the results between the computer-assisted surgical group (CASPAR) and the conventional group. METHODS: Ten femoral components were implanted either manually or by CASPAR in cadaver femurs. The specimens were cut to 3 mm thick slices. Microradiograms of every slice were sent to a computer for analysis with special software (IDL). The gaps and the medullary cavities between component and bone, the direct bone contact area of the implant surface, the gap width and the percentage of gap and bone contact area were measured in every slice. RESULTS: In the proximal implant coated with HA of the CASPAR group, the average percentage of bone contact reached 93.2% (ranging from 87.6% to 99.7%); the average gap percentage was 2.9% (ranging from 0.3% to 7.8%); the maximum gap width was 0.81 mm and the average gap width was only 0.20 mm. While in the conventional group, the average percentage of bone contact reached 60.1% (ranging from 49.2% to 70.4%); the average gap percentage was 32.8% (ranging from 25.1% to 39.9%); the maximum gap width was 2.97 mm and the average gap width was only 0.77 mm. The average gap around the implant in the CASPAR group was only 9% of that in the manual group; the maximum and average gap widths were only about 26% of those in the manual group. On the other hand, the CASPAR group showed 33% tighter bone contact than the manual group. CONCLUSION: With the use of robotics-assisted system, significant progress can be achieved for femoral preparation in total hip arthroplasty. PMID- 15362201 TI - Allograftic bone marrow-derived mesenchymal stem cells transplanted into heart infarcted model of rabbit to renovate infarcted heart. AB - OBJECTIVE: To investigate the directed transplantation of allograftic bone marrow derived mesenchymal stem cells (MSCs) in myocardial infarcted (MI) model rabbits. MATERIALS AND METHODS: Rabbits were divided into 3 groups, heart infarcted model with MSCs transplanted treatment (MSCs group, n = 12), heart infarcted model with PBS injection (control group, n = 20), sham operation with PBS injection (sham group, n = 17). MSCs labelled by BrdUrd were injected into the MI area of the MSCs group. The same volume of PBS was injected into the MI area of the control group and sham group. The mortality, LVIDd, LVIDs and LVEF of the two groups were compared 4 weeks later. Tropomyosin inhibitory component (Tn I) and BrdUrd immunohistochemistry identified the engrafted cells 4 weeks after transplantation. RESULT: The mortality of the MSCs group was 16.7% (2/12), and remarkably lower than the control group's mortality [35% (7/20) (P < 0.05)]. Among the animals that survived for 4 weeks, the LVIDd and LVIDs of the MSCs group after operation were 1.17+/-0.21 cm and 0.74+/-0.13 cm, and remarkably lower than those of the model group, which were 1.64+/-0.14 cm and 1.19+/-0.12 cm (P < 0.05); the LVEF of the MSCs group after operation was 63+/-6%, and remarkably higher than that of the model group, which was 53+/-6% (P < 0.05). Among the 10 cases of animals that survived for 4 weeks in the MSCs group, in 8 cases (80%), the transplanted cells survived in the non MI, MI region and its periphery, and even farther away; part of them differentiated into cardiomyocytes; in 7 cases (70%), the transplanted cells participated in the formation of blood vessel tissue in the MI region. CONCLUSION: Transplanted allograftic MSCs can survive and differentiate into cardiomyocytes, form the blood vessels in the MI region. MSCs transplantation could improve the heart function after MI. PMID- 15362202 TI - The diagnostic significance of the detection of cytokeratin 19 mRNA by quantitative RT-PCR in benign and malignant pleural effusions. AB - OBJECTIVE: To evaluate the diagnostic significance of detecting cytokeratin 19 (CK19) mRNA by quantitative reverse transcription polymerase chain reaction (RT PCR) in benign and malignant pleural effusions. METHODS: CK19 mRNA was examined by quantitative RT-PCR and CK19 was detected by Enzyme-linked immunoadsorbent assay (ELISA) in 32 patients with malignant pleural effusions and 35 patients with benign pleural effusions. RESULTS: On the threshold of 200 copies/microl, the positive rate of CK19 mRNA in patients with malignant pleural effusions was 62.5%. The positive rates of CK19 mRNA and CK19 in the malignant pleural effusions were significantly higher than those in the benign group (P<0.01). Furthermore, the positive rate of CK19 mRNA was higher than that of CK19 in the malignant group (P<0.05). CONCLUSION: Detection of CK19 mRNA can be a promising diagnostic marker in differential diagnosis of benign and malignant pleural effusions. PMID- 15362203 TI - Effects of pH management during deep hypothermic bypass on cerebral oxygenation: alpha-stat versus pH-stat. AB - OBJECTIVE: There is a remarkable lack of scientific evidence to support the option to use alpha-stat or pH-stat management, as to which is more beneficial to brain protection during deep hypothermic CPB. This study examined cortical blood flow (CBF), cerebral oxygenation, and brain oxygen consumption in relation to deep hypothermic CPB with alpha-stat or pH-stat management. METHODS: Twenty-two pigs were cooled with alpha-stat or pH-stat during CPB to 15 degrees C esophageal temperature. CBF and cerebral oxygenation were measured continuously with a laser flowmeter and near-infrared spectroscopy, respectively. Brain oxygen consumption was measured with standard laboratory techniques. RESULTS: During CPB cooling, CBF was significantly decreased, about 52.2%+/-6.3% (P<0.01 vs 92.6%+/-6.5% of pH stat) at 15 degrees C in alpha-stat, whereas there were no significant changes in CBF in pH-stat. While cooling down, brain oxygen extraction (OER) progressively decreased, about 9.5%+/-0.9% and 10.9%+/-1.5% at 15 degrees C in alpha-stat and pH-stat, respectively. At 31 degrees C the decreased value in pH-stat was lower than in alpha-stat (29.9%+/-2.7% vs 22.5%+/-1.9%; P<0.05). The ratio of CBF/OER were 2.0+/-0.3 in alpha-stat and pH-stat, respectively; it was kept in constant level in alpha-stat, and significantly increased by 19 degrees C to 15 degrees C in pH-stat (4.9+/-0.9 vs 2.3+/-0.4; P<0.01). In mild hypothermia, cerebral oxyhemoglobin and oxygen saturation in alpha-stat were greater than that in pH stat (102.5%+/-1.4% vs 99.1%+/-0.7%; P<0.05). In deep hypothermia, brain oxygen saturation in pH-stat was greater than that in alpha-stat (99.2%+/-1.0% vs 93.8%+/-1.0%; P<0.01), and deoxyhemoglobin in pH-stat decreased more greatly than that in alpha-stat (28.7%+/-6.8% vs 54.1%+/-4.7%; P<0.05). CONCLUSIONS: In mild hypothermic CPB, brain tissue oxygen saturation was greater in alpha-stat than in pH-stat. However, cerebral oxygenation and brain tissue oxygen saturation were better in pH-stat than in alpha-stat during profound hypothermia. PH-stat strategy provided much more oxygen to brain tissue before deep hypothermic circulatory arrest. PMID- 15362204 TI - Immunotherapy of intracranial G422 glioblastoma with dendritic cells pulsed with tumor extract or RNA. AB - OBJECTIVE: To investigate the anti-tumor efficacy of dendritic cell (DC)-based vaccines pulsed with tumor extracts or RNA in a mouse model of intracranial G422 glioblastoma. METHODS: Bone marrow-derived DCs were pulsed ex vivo with tumor extracts or RNA. Ninety female mice harboring 4-day-old intracranial G422 glioblastomas and 126 normal mice were treated with three spaced one week apart subcutaneous injections either with PBS, unpulsed DCs, G422 tumor extracts, RNA, DCs pulsed with G422 tumor extracts (DC/extract) or with RNA (DC/RNA). Seven days after the third immunization of normal mice, the spleens of 36 of them were harvested for cytotoxic T lyphocyte (CTL) assays and the others were challenged in the brain with G422 tumor cells. All the treated mice were followed for survival. Some mice brains were removed and examined pathologically when they died. RESULTS: Immunization using DC/extract or DC/RNA significantly induced G422 specific CTL responses compared with control groups (P<0.01). Vaccination with DC/extract or DC/RNA, either prior to G422 tumor challenge or in tumor-harboring mice, significantly prolonged survival compared with other control groups (P<0.01). CONCLUSION: DCs pulsed with tumor extracts or RNA derived from autologous tumors has potential antitumor effects via activation of cell-mediated immunity. Our results suggest a useful therapeutic strategy against gliomas. PMID- 15362206 TI - Duplicate publication. PMID- 15362207 TI - 25th anniversary of the Epidemiological Bulletin (EB) of the Pan American Health Organization (PAHO). PMID- 15362205 TI - Expression of matrix metalloproteinase 9 in nasopharyngeal carcinoma and association with Epstein-Barr virus infection. AB - OBJECTIVE: To evaluate the expression of matrix metalloproteinase-9 (MMP9) in nasopharyngeal carcinoma and the association between MMP9 and Epstein-Barr virus infection. METHODS: The MMP9 expression was studied by immunohistochemical analysis; and Epstein-Barr virus encoded small nuclear mRNA-1 (EBER-1) produced by in situ hybridization were examined in 41 nasopharyngeal carcinoma sections, and the relation between them, and the associations of MMP9 with clinical features were statistically analyzed. RESULTS: Positive expression rate of MMP9 was 73.17%. The expression of MMP9 showed significant positive correlation with the expression of EBER-1 (gamma=0.483, P=0.001). There was significant association of MMP9 expression with lymph nodes metastasis and clinical stage (P<0.001), non-significant association with age, gender, pathological classification and T classification. CONCLUSIONS: The highly pronounced expression of MMP9 is associated with cervical lymph nodes metastasis. Epstein Barr virus can enhance NPC metastasis by up-regulating the expression of MMP9. PMID- 15362209 TI - Avian influenza. PMID- 15362208 TI - Deaths from motor vehicle traffic accidents in selected countries of the Americas, 1985-2001. PMID- 15362210 TI - Situation of severe growth retardation in first-grade schoolchildren in countries of Central America around 2000. PMID- 15362211 TI - [Guideline for appropriate use of radioactive medicine]. PMID- 15362212 TI - On: chaotic possibilities: toward a new model of development. PMID- 15362213 TI - Changes over time in memory, processing speed and clock drawing tests help to discriminate between vascular cognitive impairment, mild cognitive impairment and Alzheimer's disease. AB - Measures of cognitive change over time may help to better discriminate between mild cognitive impairment, Alzheimer's disease and vascular cognitive impairment than single assessments. Our hypothesis was that performance in processing speed and executive function would decline with mild cognitive impairment and Alzheimer's disease. Subjects included 36 controls, 18 cases with mild cognitive impairment, eight with vascular cognitive impairment and 24 with Alzheimer's disease who were tested on a cognitive battery at two episodes with a 12-month interval. Changes in performance were determined for each group with paired means tests. Controls improved in pattern comparison speed and the CLOX, a clock drawing task to detect dysexecutive function. Those with vascular cognitive impairment declined in letter comparison speed, but improved in paragraph recall. Alzheimer's disease patients declined in CLOX and the Hopkins Verbal Learning Test. The mild cognitive impairment group showed no significant changes. Alzheimer's disease patients on treatment declined in Hopkins Verbal Learning Test, while those without treatment declined in The Placing Test and CLOX. Processing speed decline may be a marker of cerebrovascular disease, while decline in memory and executive function was more evident with Alzheimer's disease. PMID- 15362214 TI - Aid agencies facing health catastrophe in Darfur. PMID- 15362215 TI - Cases of STIs increasing in the UK, warns health agency. PMID- 15362216 TI - India reverses policy on condom advertising. PMID- 15362217 TI - China's AIDS experts worried about national AIDS programme. PMID- 15362218 TI - Researchers call for action against Anopheles gambiae mosquito. PMID- 15362219 TI - Chronic figurate skin lesions. Tinea imbricata. PMID- 15362220 TI - Elderly patients often get drugs they shouldn't, study says. PMID- 15362221 TI - Changing CPOE parameters dramatically lowers physician test ordering. PMID- 15362222 TI - Few U.S. breast surgeons do volume of operations associated with better results. PMID- 15362223 TI - Calcium and fertilization: the beginning of life. AB - The explosive increase in Ca2+ that occurs in the cytosol at fertilization is brought about by the activation of Ca2+-release channels in the intracellular stores. Inositol 1,4,5-trisphosphate (InsP3) is traditionally considered to be the messenger that initiates the increase and spreading of the activating Ca2+ wave. In line with this hypothesis, recent evidence suggests that the penetrating sperm delivers into mammalian eggs a novel isoform of phospholipase C (PLC), which promotes the formation of InsP3. By contrast, data from echinoderms studies indicate that the newly discovered second messenger nicotinic adenine dinucleotide phosphate (NAADP) promotes an initial, localized increase in Ca2+, which is then followed by the InsP3-mediated globalization of the Ca2+ wave. The mechanism by which the interacting sperm triggers the production of NAADP and subsequently that of InsP3 remains obscure. PMID- 15362224 TI - E2F target genes: unraveling the biology. AB - The E2F transcription factors are downstream effectors of the retinoblastoma protein (pRB) pathway and are required for the timely regulation of numerous genes essential for DNA replication and cell cycle progression. Several laboratories have used genome-wide approaches to discover novel target genes of E2F, leading to the identification of several hundred such genes that are involved not only in DNA replication and cell cycle progression, but also in DNA damage repair, apoptosis, differentiation and development. These new findings greatly enrich our understanding of how E2F controls transcription and cellular homeostasis. PMID- 15362225 TI - Intubation during asthma hospitalizations is associated with worse outcomes, higher costs. PMID- 15362226 TI - Capping protein: new insights into mechanism and regulation. AB - Temporal and spatial control of the actin cytoskeleton are crucial for a range of eukaryotic cellular processes. Capping protein (CP), a ubiquitous highly conserved heterodimer, tightly caps the barbed (fast-growing) end of the actin filament and is an important component in the assembly of various actin structures, including the dynamic branched filament network at the leading edge of motile cells. New research into the molecular mechanism of how CP interacts with the actin filament in vitro and the function of CP in vivo, including discoveries of novel interactions of CP with other proteins, has greatly enhanced our understanding of the role of CP in regulating the actin cytoskeleton. PMID- 15362227 TI - The talin-tail interaction places integrin activation on FERM ground. AB - Integrins are essential receptors for the development and functioning of multicellular animals because they mediate cell migration and cell adhesion, and regulate cell proliferation and apoptosis. Cellular regulation of the affinity of integrins for ligands - so-called 'integrin activation' - is a central property of these receptors. Integrin activation controls cell adhesion, migration and extracellular matrix assembly, thereby contributing to processes such as angiogenesis, tumor cell metastasis, inflammation, the immune response and hemostasis. Recent studies indicate that a crucial, final step in integrin activation is the binding of talin, a cytoskeletal protein, to the cytoplasmic domain of the integrin beta subunit. These results provide a focus for unraveling the many biochemical pathways implicated in integrin activation and suggest a general structural model for the connections between integrins and diverse cellular signal transduction pathways. PMID- 15362228 TI - 'Cap-tabolism'. AB - distinctive feature of eukaryotic mRNA and small nuclear RNA (snRNA) that are transcribed by RNA polymerase II (Pol II) is the presence of a cap structure at their 5' end. This essential modification serves as an inviting 'landing pad' for factors that are involved in various cellular processes such as pre-mRNA splicing, nucleocytoplasmic RNA export and localization, and translation initiation. Because of the important functions mediated by the mRNA cap, this structure needs to be modified and/or degraded in a tightly controlled manner. Several cellular and viral systems implicated in cap metabolism have been uncovered recently; their analyses provide interesting new information on cell structure and function. PMID- 15362229 TI - Energy transduction in transmembrane ion pumps. AB - Recent crystallographic structures of three different ion pumps provide a first view of the mechanisms by which these molecular machines transfer ions across cell membranes against an electrochemical gradient. Each of the structures reinforces the concept that several buried counter ions have central roles in substrate recruitment, substrate binding and energy transduction during ion pumping. The spatial organization of the counter ions suggests that, initially, one or more counter ions lowers the Born energy cost of binding a substrate ion in the low-dielectric interior of the membrane. Subsequently, a ligand-induced conformational change seems to close a charged access gate to prevent backflow from a subsequent, low-affinity state of the pump. A final role of the buried counter ions might be to couple the input of external energy to a small charge separation between the substrate ion and the buried counter ions, thereby decreasing the binding affinity for the substrate ion in preparation for its release on the high-energy side of the membrane. PMID- 15362230 TI - Quality care illustrates the value of EMS vs. car transport. PMID- 15362231 TI - Hydrogen cyanide: fire smoke's silent killer. PMID- 15362232 TI - Smoke inhalation & acute cyanide poisoning. Hydrogen cyanide poisoning proves increasingly common in smoke-inhalation victims. PMID- 15362233 TI - Hydroxocobalamin: treatment for smoke inhalation-associated cyanide poisoning. Meeting the needs of fire victims. PMID- 15362234 TI - Cyanide as a chemical terrorism weapon. PMID- 15362235 TI - Evaluation of renal functional reserve of contralateral kidney of two-kidney, one clip Goldblatt hypertensive rats. AB - To evaluate the presence of hyperfiltration in two-kidney, one clip (2K, 1C) Goldblatt hypertensive rats, micropuncture studies of the unclipped kidney were used to examine renal functional reserve (RFR) with glycine (G) infusion in normal (N) rats and 2K, 1C rats. Systemic hypertension in 2K, 1C rats was associated with significantly increased values of single-nephron glomerular filtration rate (SNGFR), glomerular plasma flow (QA) and glomerular capillary hydrostatic pressure (PGC) when compared with N rats. Glycine infusion produced a marked increase in SNGFR, QA and whole-kidney GFR in N rats. In contrast, no changes were observed in these parameters in 2K, 1C rats. Lack of response in 2K, 1C rats was related to the failure of renal vessels to dilate during G infusion. These results suggest that loss of renal functional reserve in systemic hypertension is dependent on an abnormal function of renal vascular resistances. PMID- 15362236 TI - [U. H. Peters: Pros and cons as to ICD-10 chapter V. Fortschr Neurol Psychiat 2003; 71(3):115-7]. PMID- 15362237 TI - Yin-yang genetics, or the HSD deCODE controversy. AB - Iceland has attracted world-wide attention because of a controversy surrounding the Icelandic Health Sector Database Act of 1998 and the genomics company deCODE Genetics. The controversy concerns limits to state power, bioethics and regulation of research, and civil liberties. Understanding it necessitates paying attention to the yin-yang-like character of deCODE; it is international or Icelandic, what is the relation between the HSD and deCODE, will the HSD engulf the Icelandic nation as a whole? PMID- 15362238 TI - Switching between science and culture in transpecies transplantation. AB - This article discusses xenotransplantation (XTP: the surgical role of nonhuman tissues, organs, and cells for human transplantation) and examines the way its scientific promoters have defended their technology against potentially damaging public representations. The authors explore the criteria used to legitimate the selection of the pig as the best species from which to "harvest" transplant tissues in the future. The authors' analysis shows that scientists and medical practitioners routinely switch between scientific and cultural repertoires. These repertoires enable such actors to exchange expert identities in scientific discourse for public identities in cultural discourse. These discourses map onto similarities and differences between animal donors and human hosts. Finally, the case is used to comment on a number of related approaches where the dynamics of medical and scientific authority are discussed. PMID- 15362239 TI - "What do you think about genetic medicine?" Facilitating sociable public discourse on developments in the new genetics. AB - An important aspect of any meaningful public discussion about developments in gene technology is the provision of opportunities for interested publics to engage in sociable public discourse with other lay people and with experts. This article reports on a series of peer group conversations conducted in late 1996 and early 1997 with sixteen community groups in Perth, Western Australia, interested in gene therapy technology. With the case of cystic fibrosis as a particular focus, and using background resource material as a stimulus for discussion, the participating groups explored a range of value issues arising from the new genetic medicine. This more discursive context enabled participants to express a number of background or life-world concerns about genetic medicine, concerns that are often obscured by the dominant biomedical and bioethical discourses. PMID- 15362240 TI - Is there a role for physicians in health and medical resource allocation? AB - Resource allocation decisions affecting health and medical care must be based on sound ethical and economic analysis. Should physicians be involved in these discussions? Some posit that physicians must be patient-advocates, and thus, cannot participate. Others argue that physicians have an integral role to play in shaping the health care system. This article proposes a role for physicians at all levels of resource allocation discussions. PMID- 15362241 TI - Medical records, 2: security and confidentiality. PMID- 15362242 TI - Professional integrity. PMID- 15362243 TI - Proposed guidelines for housestaff interaction with pharmaceutical companies. AB - The relationship between the medical profession and the pharmaceutical companies is complex, since industry depends on physicians for research and development, and for sales of its products. As a result, physician interactions with industry could undermine the patient-centered medical ethic, and jeopardize the physician patient relationship. There is evidence that physicians are influenced by their encounters with the pharmaceutical industry. Particularly troubling is the increasing interaction of pharmaceutical companies and housestaff through detailing and educational support. Several studies have documented the pervasiveness of such relationships in training programs. Although professional bodies have developed criteria to help physicians in their dealings with the pharmaceutical industry, no principles have yet been formulated to address the unique aspects of resident contact with pharmaceutical companies. Guidelines designed to answer this need are proposed in this article. PMID- 15362244 TI - Geriatric psychiatry: a subspecialty whose time has come. PMID- 15362245 TI - Canadian Outcomes Study in Dementia: study methods and patient characteristics. AB - OBJECTIVE: To describe the methods and patient characteristics of the Canadian Outcomes Study in Dementia (COSID). METHODS: COSID is a 3-year prospective study of dementia patients living in the community at the time of study registration. We assessed patients' cognition, behaviour, and functioning every 6 months, using the Modified Mini-Mental State Examination (3MS), the Neuropsychiatric Inventory (NPI), and the Functional Autonomy Measurement System (SMAF), respectively. We assessed caregivers, using the Zarit Burden Interview (ZBI). Additional information included the Global Deterioration Scale (GDS), patients' driving status, and clinical information including family history, dementia type, concomitant medications, and comorbid conditions. From the patient or caregiver, we collected details of inpatient and outpatient resources used by the patient and (or) caregiver. RESULTS: We enrolled 766 patients from 31 Canadian sites. Overall mean age was 76.8 years, and mean age of onset was 73.1 years. Of the total patients, 98% were white, 54% were women, and 84% were diagnosed with Alzheimer's disease. Mean baseline 3MS was 66.5, NPI was 9.5, and SMAF was 18.30. Of these patients, 48% reported a GDS score of 3 (that is, moderate), 16% reported a GDS score of 4 (that is, moderately severe), and the remaining 36% reported a GDS score of 1 or 2 (that is, mild or very mild). At baseline, 83% of patients received cholinesterase inhibitors, 46% received nonsteroidal antiinflammatory drugs, 39% received vitamin E, and 25% received antidepressants. Adult day care and home help were the largest cost factors in this population, with mean monthly costs of $65 and $64, respectively. We found interesting differences in the resources used among geographic regions and care settings. CONCLUSIONS: COSID is already generating valuable information about treatment patterns, outcomes, and resource use in Canadian patients with dementia. As the data mature, it will be possible to build robust models on treatment effectiveness and costs of care. PMID- 15362246 TI - Exploring the links between depression, integrity, and hope in the elderly. AB - OBJECTIVE: To explore the links between depression, integrity, and hope in the elderly. METHOD: For this pilot study, we recruited a voluntary sample of cognitively intact elderly patients receiving psychiatric care (n = 35). Recruitment sources included an inpatient geriatric psychiatry unit (n = 14), a geriatric psychiatry day program (n = 6), and an outpatient geriatric psychiatry service (n = 15). Participants completed a questionnaire designed to measure depression, acording to the Geriatric Depression Scale Short Form (GDSSF); integrity, according to the Sense of Coherence Scale-Short Form (SOC-13); and hope, according to the Hope Differential-Short (HDS) and the Hope Numerical Rating Scale (Hope-NRS). The HDS consists of 3 separate subscales: Personal Spirit, Risk, and Authentic Caring. We analyzed the data, using descriptive statistics, t tests, and Pearson correlations. RESULTS: Patients with no depression (n = 17) showed a greater sense of coherence (SOC) (P < 0.01), higher levels of hope (Hope-NRS, P < 0.05), enhanced Personal Spirit (HDS subscale, P < 0.05), and greater risk taking (HDS subscale, P < 0.01) than did patients with depression (n = 18). The 2 variables that correlated most highly with depression were SOC (r = -0.65, P < 0.01) and Risk (HDS subscale, r = -0.62, P < 0.01). CONCLUSIONS: These findings suggest that depression, integrity, and hope are highly interrelated in the elderly population and may influence mastery of the developmental tasks of aging. Further research is warranted to better understand these complex experiences in late life. PMID- 15362247 TI - Driving and dementia in Ontario: a quantitative assessment of the problem. AB - BACKGROUND: The population is becoming increasingly aged, and concomitantly, the prevalence of dementia is steadily rising. Persons aged 65 years and over are likely to continue driving for many years and often well into the dementia process. METHODS: Ontario Ministry of Transportation driving data, census data, and dementia prevalence data were combined to determine the number of persons with potential dementia who are driving, both now and in about 25 years' time. RESULTS: Actual and projected Ontario figures show that the number of senior drivers will increase markedly from just under 500,000 in 1986 to nearly 2,500,000 in 2028. Similarly, the number of drivers with dementia is also increasing. Although not all drivers with dementia are necessarily dangerous, most are estimated to continue driving well into the disease process. By combining the above-mentioned data sets, a best estimate of the number of drivers with dementia in Ontario was derived. It is estimated that this group has grown from just under 15,000 in 1986 to about 34,000 in 2000 and will number nearly 100,000 in 2028. INTERPRETATION: Increasingly, the responsibility for identifying drivers with dementia has fallen on the health care system, a role for which it was never designed nor equipped to handle. The risks associated with the dramatically increasing number of drivers with dementia demand a psychometrically sensitive and efficient screening procedure. PMID- 15362248 TI - GABAergic function in Alzheimer's disease: evidence for dysfunction and potential as a therapeutic target for the treatment of behavioural and psychological symptoms of dementia. AB - Alzheimer's disease (AD) is characterized by disruptions in multiple major neurotransmitters. While many studies have attempted to establish whether GABA is disrupted in AD patients, findings have varied. We review evidence for disruptions in GABA among patients with AD and suggest that the variable findings reflect subtypes of the disease that are possibly manifested clinically by differing behavioural symptoms. GABA, the major inhibitory neurotransmitter, has long been a target for anxiolytics, hypnotic sedatives, and anticonvulsants. We review the clinical use of GABAergic agents in treating persons with AD symptoms. While newer generation GABAergic medications are now available, they have yet to be evaluated among patients with AD. PMID- 15362249 TI - Surrogate decision making: special issues in geriatric psychiatry. AB - Surrogate decision-makers, usually family and friends, are often called on to assist seniors who are incapable of making certain decisions. The literature to date has focused primarily on decisions regarding medical treatment. Less has been written about issues faced by surrogates when making other types of decisions that often arise in the practice of geriatric psychiatry. This article outlines a case relating to personal care decision-making. The discussion focuses on areas that need to be addressed to understand and assist surrogates in making personal care decisions in the future. PMID- 15362250 TI - Defining best practices for specialty geriatric mental health outreach services: lessons for implementing mental health reform. AB - OBJECTIVE: This paper reviews fundamental areas that inform best practices for outreach services as they relate to shared care, education, and program and systems development and illustrates a framework for defining best practices. METHOD: We examined literature that addresses the needs and characteristics of older persons with mental illness, studies that investigate geriatric mental health outreach services, and relevant theoretical developments. In addition, we synthesized some of the health services field research that we conducted. RESULTS: This analysis provided a foundation for understanding the processes required to identify and implement best practices in specialty geriatric mental health outreach. Constructive insights for a comprehensive approach to the development, implementation, and evaluation of these services emerged. CONCLUSIONS: The needs of an aging population, the articulation of principal functions for specialty services within mental health reform, and the realities of individual communities converged to establish direction for service provision. The use of best practices across a range of health and social services, combined with an effective process that facilitates the exchange of knowledge between sectors, establishes an important mechanism by which shared care, education, and program and systems development can be achieved. PMID- 15362251 TI - Preventing postpartum depression part I: a review of biological interventions. AB - OBJECTIVE: This paper critically reviews the literature to determine the current state of scientific knowledge concerning the prevention of postpartum depression (PPD) from a biological perspective. METHODS: The criteria used to evaluate the interventions were derived from the standardized methodology developed by the Canadian Task Force on Preventive Health Care. Databases searched for this review include Medline, PubMed, Cinahl, PsycINFO, Embase, ProQuest, the Cochrane Library, and the World Health Organization Reproductive Health Library. Studies selected were peer-reviewed English-language articles published between January 1, 1966, and December 31, 2003. RESULTS: Seven studies that met criteria were examined. These studies focused on evaluating the preventive effect of antidepressant medication, estrogen and progesterone therapy, thyroid therapy, docosahexanoic acid, and calcium supplementation. Although some of these interventions have been examined rigorously for depression unrelated to childbirth, methodological study limitations render intervention efficacy equivocal for PPD; thus, there is limited strong evidence to guide practice or policy recommendations. CONCLUSIONS: Despite the recent upsurge of interest in this area, many questions remain unanswered, which has several implications for research. PMID- 15362252 TI - Suicidal ideation in inpatients with acute schizophrenia. AB - OBJECTIVE: Schizophrenia has been associated with a high rate of suicide. This study investigates the prevalence of suicidal ideation in a population of inpatients with acute schizophrenia, together with the clinical parameters associated with suicidal thoughts. METHOD: We assessed 93 schizophrenia patients. We matched subjects for age and sex and compared subjects with and without suicidal thoughts. We performed stepwise multiple regression analysis to assess the association between specific clinical symptoms and suicidal ideation. RESULTS: Of the patients, 20.4% reported suicidal thoughts during the last 15 days. Severity of depressive symptoms, motor retardation, guilt feelings, pathological guilt, and self-depreciation predicted the patients' suicidal ideation. CONCLUSIONS: Suicidal thoughts are frequent among inpatients with acute schizophrenia. Prevention of suicidal behaviour should include helping patients improve their self-esteem and reducing depression and guilt feelings. PMID- 15362253 TI - The RCPSC oral examination: patient perceptions and impact on participating psychiatric patients. AB - OBJECTIVE: To investigate patient perceptions and the impact of participating in the Royal College of Physicians and Surgeons of Canada (RCPSC) long case oral examination. METHOD: Immediately following the RCPSC exam, participating psychiatric patients completed a Participant Impact Questionnaire. We combined results for 2002 and 2003 from both the western and eastern sites. RESULTS: Overall, there was a 93% response rate (132/142): 98% (65/66) for the western site and 88% (67/76) for the eastern site. Of the respondents, 94% said they would participate in the RCPSC oral exam again, if required. Most (92%) either agreed or strongly agreed that the exam was well organized; 76% either disagreed or strongly disagreed that the exam was stressful; only 9% agreed that they participated to earn extra money; and 82% strongly disagreed that they felt pressured by their psychiatrist to participate. Patients' written reasons for participating included, but were not limited to, helping to train future psychiatrists. We briefly discuss aspects of the use of real patients vs standardized patients in the RCPSC oral exam. CONCLUSION: Patients who participated in the RCPSC long case oral exam generally found this experience very positive. The exam did not appear to have a negative impact on patients. PMID- 15362254 TI - Symptoms defined by parents' and teachers' ratings in attention-deficit hyperactivity disorder: changes with age. AB - OBJECTIVE: To determine whether Child Behavior Checklist/4-18 (CBCL) and Teacher Report Form (TRF) scores of children and adolescents with a first-time diagnosis of attention-deficit hyperactivity disorder (ADHD) are different and whether there is a similar difference in normal control subjects. METHOD: We analyzed the CBCL and TRF scores of 146 patients (124 boys and 22 girls, aged 6 to 18 years; mean age 11.0 years, SD 3.6). We analyzed the same scores for 274 age and sex matched control subjects recruited from a nationally representative sample. RESULTS: Subjects with ADHD had significantly higher CBCL and TRF scores than control subjects. Age was significantly correlated with scores on the CBCL and TRF subscales Social Withdrawal, Somatic Complaints, and Internalization Problems; with scores on the CBCL subscale Attention Problems; and with scores on the TRF subscale Anxiety-Depression. In the group with ADHD, age was negatively correlated with scores on the CBCL and TRF subscale Externalizing Problems and with scores on the TRF subscale Aggressive Behavior. In the control group, the only significant correlation was between age and the CBCL subscale Somatic Complaints score. CONCLUSIONS: These results indicate that underdiagnosis of ADHD in childhood may cause the emergence of greater internalization problems in adolescence. PMID- 15362255 TI - Mirtazapine-induced shopping spree. PMID- 15362256 TI - Age at onset of bipolar II disorder. PMID- 15362257 TI - Venlafaxine-associated hypomania in unipolar depression. PMID- 15362258 TI - Hypnopompic hallucinations during olanzapine treatment. PMID- 15362259 TI - Atypical neuroleptic malignant syndrome caused by clozapine and venlafaxine: early brief treatment with dantrolene. PMID- 15362260 TI - A case of de Clerambault syndrome in a male stalker with paranoid schizophrenia. PMID- 15362261 TI - Calcitonin treatment for phantom limb pain. PMID- 15362262 TI - The use of atomoxetine adjunctively in fibromyalgia syndrome. PMID- 15362263 TI - Re: Autism--its detection, causes, and treatment. PMID- 15362264 TI - [Risk factors for arterial disease]. AB - The risk factors of arterial disease (FREA) predict a future damage over the vascular system of the human body. Its detection are considered a key for the diagnostic as well as for the preventive and even curative strategies. For a long time, scientist considered those factors originated as a consecuence of large studies during the middle of the last century, with current validity up to our days. A simple classification spoke of them as traditionals. Further investigations described the so called new or emergents.factors that where joint together accordingly to their actions: coagulation factors, psicosocial, inflamatories and infectious. A recent classification, taking into account the type of impact, divided them into; causatives, predisposals and conditionals. Also, it was described a mechanism, the oxidative power, with consecuences over the endothelium, in the last part of the process. Before, another mechanism was described: the insulin resistance and the hiperinsulinism, bases for the Metabolic Syndrome, that includes a number of traditional risk factors. PMID- 15362265 TI - [Annex: Prevalence of insulin resistance and its association with type 2 diabetes and other cardiovascular risk factors]. AB - We present the results of an epidemiological research undertaken in the province of Cordoba (Argentina) with the aim of studying insulin-resistance (IR) in relation to the increased risk for diabetes and its association with risk factors of arterial disease (RFAD). Overall, there were 1413 subjects between 20 and 70 years of age, from both sexes (Dean Funes 18885 inhabitants; sample 715; Oncativo 13880 inhabitants; sample 696). We studied traditional arterial risk factors (hypertension, diabetes, dyslipidemia, obesity, sedentary life, smoking). To evaluate IR we used the homeostasis model assessment (HOMA-IR). We found a high prevalence of IR clearly related to the traditional (RFAD), thus placing these populations, representatives of national ones, among the western populations at high risk for arterial disease. PMID- 15362266 TI - [From risk factors of arterial disease to atheroma plaque detection of the high risk plaque]. AB - The atherosclerosis is a pathologic phenomena which is caracterized by the stiffnes and enlargement of the arterial walls due to the deposit of lipid carbohidrates, blood products and calcium into the subendothelial space. The risk factors of arterial diseases makes their impact in all arteries of the human body. It is clear to day that the composition of the plaque, is the major determinant for its risk of rupture and for its logic consequence: thrombosis. Following plaque rupture, lipidic core and its high content of factors, gives a powerful substrate for the activation of the coagulation cascade. It has been significant advances in the comprehension of the mechanism involved in the development of this disease and there is a large number of evidences that the composition of the plaque is a major determinant factor in the beginning and in the severity of the acute coronary syndrome. In consequence, the capacity to identify the above mentioned composition, monitoring its progression, are valuable information for the development of acute coronary syndrome. The conventional angiocoronariography fails to predict acute episodes, in such a way that it has lost the value as a gold standard for the detection of this syndrome and this failing promotes researchers to develop new methods, invasive and non invasive to recognize the higly risk atherosclerotic plaque and the sick areas for ideal and rapid interventions. PMID- 15362267 TI - [Arterial disease. Prevention]. AB - Human beings may have or may have not risk factors for arterial disease. Those having these factors may or may not suffer from acute episodes of aterothrombosis. Those who suffer it, may have one or more risk factors for arterial disease. For this long list of situations, there are strategies with some common points. The prevention is of high priority, specially the primary one aim to those without FREA, in particular the children. It has been demonstrated the early beginning of the disease, that is why education is of great value and must be extended to all groups of ages. In some circumstances, also on early ages, it is wise to investigate the clinic and laboratory for FREA, specially in those with hereditary overload. The common measures: healthy diet, no tobacco, reduction of alcohol intake, sporting, should be incorporated in our society as a general practice. In those with FREA, adults in general, it is convenient to establish the global risk in order to be able to consider the deepness of medical intervention and the corresponding patient adherence to medical council and prescription. For those who already suffered an acute episode, the secondary prevention implicate: counselling over the factors that are able to be modified, prescription, such as platelet anti adhesive drugs and those corresponding to each individual factor. The modern tendency is the use of drug to reduce cholesterol and other lipids, anti hypertension and others pharmaceutical interventions. All members of modern society should play a role on the above mentioned preventive actions. PMID- 15362268 TI - Performance of a digital PCO2/SPO2 ear sensor. AB - OBJECTIVE: For determining the adequacy of ventilation, conventional pulse oximetry should be amended by PaCO2 (= arterial carbon dioxide partial pressure). This study investigates the precision of carbon dioxide measurements of the first digital ear-clip sensor providing continuous non-invasive monitoring of PaCO2, SpO2 (= functional arterial oxygen saturation as estimated with a pulse oximeter) and pulse rate and compares it to two conventional analog oximeters. METHODS: 30 hypoxemia episodes in 6 adult volunteers were investigated in a standardized protocol. EQUIPMENT: Masimo analog finger sensor, Nellcor analog ear sensor, SenTec digital ear sensor. RESULTS: The difference between PCO2 data (= PaCO2 estimated from the measured PcCO2 based on an algorithm by Severinghaus) (PcCO2 = cutaneous carbon dioxide pressure) and the PaCO2 is clinically unimportant. Therefore, we suggest, the two methods of estimating patient's carbon dioxide status can be used interchangeably. CONCLUSIONS: Combined digital SpO2/ PcCO2 ear sensors are very promising to allow for a fast and reliable monitoring of patient's oxygenation, hyper-/hypocapnia and ventilation with one single non invasive probe. Optimal primary signal processing--amplification and digitalisation within the probe--allow for fast and reliable downstream signal processing algorithms. The resulting short SpO2 response times give the medical staff more time to take appropriate actions. PMID- 15362269 TI - An on line technique to detect cardiac output variations and cardiovascular performances during abdominal aortic surgery. AB - Cardiac output (CO) is a parameter usually requested to assess hemo dynamic status of patient and efficacy of therapy especially in critically ill patients. This paper, in line with other research activities and new market availability, intends to correlate a parameter derived from data already acquired in standard patient monitoring (systemic arterial pressure--Pas) in order to identify CO trends and, more generally, to obtain information on the efficiency of cardiovascular system of the patient in examination. Attention has been focussed on patients undergoing abdominal aneurysm aortic (AAA) surgery with combined anaesthesia (epidural and light general). Awake correlation was found between maximum value of Pas time derivative, max (dPas/dt), and CO measured with thermodilution method on 56 measurements on 10 patients. To evaluate further diagnostic capability of max (dPas/dt), we compare its trend with other parameters but no statistical significant results have been obtained. Therefore the target parameter can be profitably used in the examined patients to monitor CO trend and, in correlation with other parameters, as a sign of efficiency of the cardio circulatory system. PMID- 15362270 TI - The sounds of saturation. AB - Audible pulse tones, based on a variable-pitch frequency scale, allow the anesthesiologist to determine the patient's oxygen saturation without constant visual examination of the monitor display. The ability to reliably detect oxygen saturation levels based on audible pulse tones may be compromised when multiple pulse oximeter systems are used. The goal of this observational study was to examine the pitch frequency scales from several different pulse oximeter manufacturers. Using simulator technology, incremental oxygen saturations between 80% and 100% were created. The frequencies of various pulse tones in this range were measured with an oscilloscope. From this data, the relationship between oxygen saturation and corresponding pulse tone frequency was examined. Diagrammed results showed wide variation in the pulse frequency scales used by the pulse oximeters tested. At any oxygen saturation level between 80% and 100%, none of the monitors had the same pulse tone frequency. With such variation among systems, the ability to accurately determine oxygen saturation from a pulse tone may be hindered. In locations where different pulse oximeter systems are encountered, the potential for confusion exists. Anesthesiologists need to be aware of these differences, and should familiarize themselves with the audible frequency scale of a particular pulse oximeter model before its use. PMID- 15362271 TI - Intramyocardial oxygen monitoring in coronary artery bypass surgery. AB - OBJECTIVE: In coronary artery bypass surgery various parameters have been used to monitor patients clinical status. Direct monitoring of myocardial oxygenation can be performed by measuring intramyocardial partial oxygen tension pressure (p ti O2). This study was performed to determine the perioperative time course of this parameter in correlation to standard monitoring parameters. METHODS: Twenty-three patients underwent standard coronary artery bypass grafting (CABG). A special polarographic microprobes was inserted into the myocardium in the distribution zone of the left anterior descending artery which was one of the target vessels of myocardial revascularization. Intramyocardial p ti O2 was monitored intra- and up to 12 hours postoperatively. Values were correlated to hemodynamic, oxygenation and procedure associated parameters. RESULTS: Myocardial oxygenation during CABG is characterized by a significant decrease of p ti O2 during cross clamping and a significant increase after removal of the cross-clamp. The postoperative time course of p ti O2 shows a steady increase of p ti O2 in the first 12 postoperative hours investigated. Preoperative ejection fraction as well as cardio-pulmonary bypass time does not seem to have an influence on the postoperative p ti O2 in these patients. Various standard monitoring parameters show complex influence on intramyocardial p ti O2- CONCLUSIONS: Determination of intramyocardial partial oxygen pressure in patients undergoing bypass surgery shows characteristic changes. Changes in p ti O2 as a direct online parameter of myocardial oxygenation occur immediately after procedures that influence myocardial perfusion and therefore, may help to detect potential complications earlier than standard monitoring parameters in cardiac surgery. PMID- 15362272 TI - Evaluation of impedance based indices of cardiac contractility in dogs. AB - OBJECTIVE: Cardiac function can be monitored simply and safely by the impedance method. A number of parameters that reflect cardiac contractility can be derived from the impedance waveform. These include the systolic time ratio (STR), the index of contracticity (IC), the acceleration index (ACI) and the Heather index. This study evaluates their reliability. METHODS: In sixteen anaesthetized dogs an ultrasonic flow probe was placed on the ascending aorta using a left thoracotomy approach and catheter placed in the femoral artery to measure blood pressure. This allowed the reference measurement of cardiac contractility from blood flow (dF/dt(max)) and pressure (dP/dt(max)). Comparative 1-minute impedance based measurements were made by a RheoCardioMonitor (ACMA, Singapore), whilst contractility was increased 2 to 6 fold, over 113 (52 to 212) minutes, using dopamine and adrenaline infusions. The association between the reference and impedance measurements was determined by correlation. The correlation coefficients (r) were compared using paired t-tests. Results are presented as mean +/- SD. RESULTS: The ACI (r = 0.76 +/- 0.13) and Heather index (r = 0.74 +/- 0.14) were more closely associated with the reference measurement (dF/dt(max)) than IC (r = 0.65 +/- 0.23) (p < 0.05) and STR (r = 0.33 +/- 24) (p < 0.01). Results for ACI and the Heather index were similar. STR was unrelated to the reference method in 10 out of 16 experiments. Correlation was better when using flow probe data (dF/ dt(max)) than arterial pressure data (dP/dt(max)) (p < 0.05). CONCLUSIONS: ACI and the Heather index were the most reliable impedance derived indices of cardiac contractility. PMID- 15362273 TI - A dual closed-loop control system for mechanical ventilation. AB - OBJECTIVE: Closed-loop mechanical ventilation has the potential to provide more effective ventilatory support to patients with less complexity than conventional ventilation. The purpose of this study was to investigate the effectiveness of an automatic technique for mechanical ventilation. METHODS: Two closed-loop control systems for mechanical ventilation are combined in this study. In one of the control systems several physiological data are used to automatically adjust the frequency and tidal volume of breaths of a patient. This method, which is patented under US Patent number 4986268, uses the criterion of minimal respiratory work rate to provide the patient with a natural pattern of breathing. The inputs to the system include data representing CO2 and O2 levels of the patient as well as respiratory compliance and airway resistance. The I:E ratio is adjusted on the basis of the respiratory time constant to allow for effective emptying of the lungs in expiration and to avoid intrinsic positive end expiratory pressure (PEEP). This system is combined with another closed-loop control system for automatic adjustment of the inspired fraction of oxygen of the patient. This controller uses the feedback of arterial oxygen saturation of the patient and combines a rapid stepwise control procedure with a proportional integral-derivative (PID) control algorithm to automatically adjust the oxygen concentration in the patient's inspired gas. The dual closed-loop control system has been examined by using mechanical lung studies, computer simulations and animal experiments. RESULTS: In the mechanical lung studies, the ventilation controller adjusted the breathing frequency and tidal volume in a clinically appropriate manner in response to changes in respiratory mechanics. The results of computer simulations and animal studies under induced disturbances showed that blood gases were returned to the normal physiologic range in less than 25 s by the control system. In the animal experiments understeady-state conditions, the maximum standard deviations of arterial oxygen saturation and the end-tidal partial pressure of CO2 were +/- 1.76% and +/- 1.78 mmHg, respectively. CONCLUSION: The controller maintained the arterial blood gases within normal limits under steady-state conditions and the transient response of the system was robust under various disturbances. The results of the study have showed that the proposed dual closed-loop technique has effectively controlled mechanical ventilation under different test conditions. PMID- 15362274 TI - HICOPS: human interface computer program in space. AB - BACKGROUND: During long experimental set ups, a protocol book usually guides cosmonauts. This is not very easy to work with in microgravity conditions and is not very efficient. For the cardiovascular physiology experiment CARDIOCOG during the Belgian Soyuz Mission (Odissea, November 2002) we developed a software program that guided the cosmonauts through the experiment. The software was developed in LabVIEW, thoroughly tested by CNES and the Russian space authorities and transported to the ISS as a stand-alone application. An adapted version was used during the Spanish Cervantes Mission in October 2003. RESULTS: This program provided several advantages: (1) error procedures could be easily dealt with in using the program's incorporated error structure; (2) the experimental sequences were easy to follow for the cosmonauts; (3) the experimental duration was exactly the same for all repetitions of the experiment, since the program imposed the timing; (4) after the flight, we were able to reconstruct all sequences of the experiment using a log-file that was automatically created during the different steps of the experiment; and (5) we were able to impose exact breathing frequencies to the cosmonauts using a visual aid. CONCLUSION: Less training was necessary for the cosmonauts to learn the experiment. Reconstruction of the experiment timing was easy. Exact breathing frequencies were obtained at each repetition. The program HICOPS worked to the overall satisfaction of the cosmonauts and they preferred working with HICOPS instead of with paper flow sheets. Data for the cardiovascular experiment during both missions were obtained in a standardised way. PMID- 15362276 TI - Early evaluation and resuscitation of the pediatric trauma patient. AB - Trauma is the leading case of death for children in the United States. Effective initial resuscitation of pediatric trauma patients can reduce mortality. Guidelines have been developed to facilitate patient care in a systematic and productive manner. Advances have been made in both diagnostic and therapeutic methods. The evaluation and treatment of trauma patients will continue to engage pediatric surgeons as efforts in trauma prevention become more successful. PMID- 15362275 TI - Monitoring of arterial stiffness indices by applanation tonometry and pulse wave analysis: reproducibility at low blood pressures. AB - OBJECTIVE: Aortic pulse wave analysis (PWA) reveals valuable information related to several hemodynamic characteristics mainly in normotensive and hypertensive patients. The main indices determined by PWA are augmentation index (AI) and reflection time index (RTI), which provide an indirect estimate of arterial stiffness and pulse wave velocity. The objective of the present study was to assess the reproducibility of aortic AI and RTI obtained by an automated and commercially available system (SphygmoCor) applied in patients with low blood pressures where such data are lacking. METHODS: The study population consisted of 19 patients with cardiogenic shock due to acute myocardial infarction who underwent mechanical assistance with intraaortic balloon pump. Aortic pressure waveforms were derived from peripheral waveforms--recorded by applanation tonometry of the radial artery--by applying generalized transfer functions. On every occasion, a well-trained operator performed two measurements with 2 min interval. Multiple pairs of measurements were obtained per patient in order to study a wide range of different hemodynamic conditions. Thus, a total of 91 pairs of measurements were performed and analyzed using Bland-Altman plots. RESULTS: AI and RTI ranged within 30-184% and 10-27%, respectively. Within-observer difference was 0.10 +/- 5.82% for aortic AI and 0.14 +/- 1.2% for RTI. CONCLUSION: Pulse wave analysis and radial artery tonometry can be used to measure AI and RTI with satisfactory reproducibility even in low blood pressures. Ongoing research is required to establish PWA utility in clinical practice especially at patients with low blood pressures. PMID- 15362277 TI - Diagnosis and management of pediatric closed head injury. AB - Closed head injury in the pediatric population accounts for almost half of all new cases of traumatic brain injury. The incidence of severe brain injury appears to be less in children as compared to the adult population. Over the past several years, advances in recognition and treatment of traumatic brain injury have led to improved outcomes in both children and adults. It is imperative, therefore, for the pediatric surgeon dealing with head trauma to have an understanding of the common brain injuries in the pediatric population, their early recognition and initial management. In this article, early diagnosis and initial management of the most common forms of pediatric closed head injury are reviewed. PMID- 15362278 TI - Penetrating trauma in children. AB - Penetrating injuries account for 10% to 20% of all pediatric trauma admissions at most centers. Gunshot wounds are responsible for the overwhelming majority of penetrating traumatic injuries and have a significantly higher mortality rate than do blunt injury mechanisms. The management of penetrating injuries can be quite challenging and often requires rapid assessment and intervention. Specific management principles are guided by the anatomic location of injury, the determination of trajectory, and the suspected organs injured. Management approaches have been adopted in large part from the more robust adult experience. However, application of these strategies to similar life-threatening injuries in the pediatric population appears appropriate. PMID- 15362279 TI - The diagnosis and management of children with blunt injury of the chest. AB - Thoracic trauma remains a major source of morbidity and mortality in injured children, and is second only to brain injuries as a cause of death. The presence of a chest injury increases an injured child's mortality by 20-fold. Greater than 80% of chest injuries in children are secondary to blunt trauma. The compliant chest wall in children makes pulmonary contusions and rib fractures the most common chest injuries in children. Injuries to the great vessels, esophagus, and diaphragm are rare. Failure to promptly diagnose and treat these injuries results in increased morbidity and mortality. PMID- 15362280 TI - Blunt injury to solid abdominal organs. AB - Injuries to the solid abdominal viscera (spleen, liver, kidney, pancreas) are common in children sustaining trauma by blunt mechanisms. Success with nonoperative management of these injuries has led to recent extensions of this approach to the management of higher-grade more complicated injuries typically treated operatively. This review will discuss the current status of evaluation, management and outcome of children sustaining blunt injury to solid abdominal organs. PMID- 15362281 TI - Hollow viscous injury in the pediatric patient. AB - Injuries to the gastrointestinal tract account for 1% to 15% of intraabdominal injuries in children. Most hollow visceral injuries occur following some form of blunt trauma and motor vehicle accidents remain the most common mechanism of injury. The diagnosis of blunt intestinal injury is difficult and often delayed. Current imaging modalities are imprecise and contribute to delay. Delay is associated with morbidity and mortality in both children and adults, but the length of delay remains controversial. The purpose of this review is to examine the current diagnosis and management of hollow visceral injury in children. PMID- 15362282 TI - Pediatric orthopedic trauma: principles of management. AB - Musculoskeletal injuries in the pediatric population are unique and require a thorough evaluation by a trained specialist. Unlike adults, many of the injuries may be treated closed due to the amazing growth and remodeling potential of children. Special consideration should be taken in treating certain fracture patterns to prevent the long-term consequences of growth deformities and protect children from child abuse. It is the goal of this article to outline common orthopedic injuries in the pediatric population to facilitate proper care in the multidisciplinary evaluation and treatment of children. PMID- 15362283 TI - The national pediatric trauma registry: a legacy of commitment to control of childhood injury. AB - The National Pediatric Trauma Registry represents almost 15 years of effective collaboration among hospitals committed to improving care for the injured child. Its design of providing a "physiologic snapshot" of the injured child on presentation has supported numerous studies that have helped define the epidemiology of childhood injury and refine principles of management. Global analysis of the 103,434 records included in this database suggest that mortality is significantly higher in the very young, that vehicular injury remains a major pediatric public health challenge, and that shock is just as devastating in the child as the adult. Based on this foundation of collaborative commitment, future versions of a pediatric trauma database must harness the emerging internet technology that combines information accrual with human thought, and must extend this effort to include all the children of our world. PMID- 15362284 TI - Preventing injury and injury-related disability in children and adolescents. AB - Injury is the leading cause of death and a prevalent source of disability and excess health expenditures in children and adolescents. There are predictable patterns to injury that provide clues to prevention. Epidemiologically-based theoretical frameworks are available to guide development of injury prevention strategies, to add structure to our observations, and focus to our prevention activities. While all-cause injury mortality rates have decreased in children and adolescents over the last 20 years, large ethnic disparities persist, indicating the need for intensified efforts in high-risk communities. Strong leadership from pediatric surgeons and pediatricians operating hospital-based community injury prevention programs has produced successful reductions in child and adolescent injury rates in resource-limited and minority neighborhoods. Among the program features considered essential are: (1) an ongoing injury surveillance system; (2) well-focused, multifaceted prevention activities, including both passive and active prevention approaches; (3) education; (4) enlistment of other health professionals, local government officials, community leaders, and the public; and (5) evaluation and refinement of prevention activities. PMID- 15362285 TI - Primate models for human immunodeficiency virus infection. Evolution of receptor use during pathogenesis. AB - Animal models greatly facilitate understanding of transmission, pathogenesis and immune responses in HIV and SIV infection and provide models for studies on the effect of candidate drugs or vaccines. However, there are several aspects that one should consider when drawing conclusions from results obtained from animal models. First, the genetic relationship of primate lentiviruses cannot be disregarded because it is known that HIV-1 is more closely related to SIV of chimpanzee origin (SIVcpz) than to SIV from sooty mangabey (SIVsm) origin. Nevertheless, SIVsm and SIVmac are the ones most often used as model systems. Second, there are differences in the biological properties, like CXCR4 use and CD4-independent coreceptor use, of HIV and SIV. These differences might be relevant in virus transmission, pathogenesis and in evoking immune responses. Third, in vivo and in vitro selection may influence the results. Neutralizing antibodies may play a role in selection of variant viruses since neutralization sensitive, CD4-independent SIVsm variants seemed to be suppressed in animals that mounted a neutralizing antibody response. It is tempting to speculate that neutralizing antibodies shape the SIV/HIV infection by selecting variants with a more "closed" envelope conformation with consequences for both receptor binding and neutralization sensitivity. The SIV/monkey model, although it has important advantages, may not answer all questions asked about HIV-1 infection in human. PMID- 15362286 TI - Lead induced modulation of splenic macrophage responses on humoral and cell mediated immunity. AB - The heavy metal lead is an environmental toxic material that can induce pathophysiological changes in many organ systems. Previous studies have shown the effects of lead exposure on immune cells in different experimental animals, however, the mechanism of their influence on the immune system is unclear. We reported that in vivo lead exposure inhibits phagocytosis, nitric oxide release, induces DNA fragmentation suggesting the apoptotic death of the target cell. We have also presented evidence that inhibition of macrophage functional responses implicated alteration of humoral and cell mediated immunity. In vivo exposure to lead acetate alters the phagocytic capacity of splenic macrophages as evident from the reduction of phagocytic index of control from 19,792+/-1385.69 to 8893+/ 893 in the treated group. The amount of nitric oxide released by the control cell 2.25+/-0.125 microM is also reduced to 1.9375+/-0.0625 microM upon in vivo lead treatment. Functional integrity of the target cell is also decreased after lead exposure as obtained from the percentage of DNA fragmentation. Control group shows 33.29+/-0.11% of fragmented DNA, which is enhanced to 42.43+/-0.725% following the lead treatment. A greater percentage of DNA fragmentation upon lead treatment probably indicating that the heavy metal induces apoptosis. The humoral immune response is also altered after lead exposure as indicated by the decrease of the antibody titre in control group from 1:2048 to 1:128 in the treated group. From the DTH reaction, it was observed that the mean diameter of swollen foot pad of control mice is 0.329+/-0.15 cm and that of lead treated mice is 0.274+/-0.056 cm. It can, therefore, be suggested that lead inhibits normal functional activities of splenic leukocytes, particularly phagocytosis and also affects the functional integrity of cells by inducing DNA fragmentation. The study may demonstrate the usefulness of investigation of humoral immune system and leukocyte functions as sensitive parameters in detecting the effects of lead toxicity. PMID- 15362287 TI - A comparative study on phyllosphere nitrogen fixation by newly isolated Corynebacterium sp. & Flavobacterium sp. and their potentialities as biofertilizer. AB - A number of nitrogen fixing bacteria has been isolated from forest phyllosphere on the basis of nitrogenase activity. Among them two best isolates are selected and identified as Corynebacterium sp. AN1 & Flavobacterium sp. TK2 able to reduce 88 and 132 n mol of acetylene (10(8)cells(-1)h(-1)) respectively. They were grown in large amount and sprayed on the phyllosphere of maize plants as a substitute for nitrogenous fertilizer. Marked improvements in growth and total nitrogen content of the plant have been observed by the application of these nitrogen fixing bacteria. An average 30-37% increase in yield was obtained, which is nearer to chemical fertilizer treatment. Comparatively better effect was obtained by application of Flavobacterium sp. PMID- 15362288 TI - Properties of an immobilized lipase of Bacillus coagulans BTS-1. AB - Lipase (EC 3.1.1.3) is a tri-acylglycerol ester hydrolase, catalysing the hydrolysis of tri-, di-, and mono-acylglycerols to glycerol and fatty acids. To study the effect of adsorption of a lipase obtained from Bacillus coagulans BTS 1, its lipase was immobilized on native and activated (alkylated) matrices, i.e. silica and celite. The effect of pH, temperature, detergents, substrates, alcohols, organic solvent etc. on the stability of the immobilized enzyme was evaluated. The gluteraldahyde or formaldehyde (at 1% and 2% concentration, v/v) activated matrix was exposed to the Tris buffered lipase. The enzyme was adsorbed/entrapped more rapidly on to the activated silica than on the activated celite. The immobilized lipase showed optimal activity at 50 degrees C following one-hour incubation. The lipase was specifically more hydrolytic to the medium C length ester (p-nitro phenyl caprylate than p-nitro phenyl laurate). The immobilization/entrapment enhanced the stability of the lipase at a relatively higher temperature (50 degrees C) and also promoted enzyme activity at an acidic pH (pH 5.5). Moreover, the immobilized lipase was quite resistant to the denaturing effect of SDS. PMID- 15362289 TI - Triflupromazine: a microbicide non-antibiotic compound. AB - The antipsychotic phenothiazine triflupromazine, possessing a methyl-thio substituent at position 10 and a fluorine moiety at position 2, exhibited significant antibacterial activity against 279 strains of Gram-positive and Gram negative bacteria. The minimum inhibitory concentration (MIC) of the drug, according to the agar dilution method, was between 2 and 50 microg/ml for Staphylococcus aureus, and 5 and 100 microg/ml for shigellae and vibrios. Triflupromazine, when injected intraperitoneally into Swiss albino mice at a concentration of 30 microg/mouse (20 g), manifested a significant protection to the mice (p<0.001) when they were challenged with 50 median lethal dose (MLD) of Salmonella typhimurium NCTC 74. Moreover, there was a statistically significant reduction in the number of viable bacteria in organ homogenates and blood of mice treated with this phenothiazine compound. PMID- 15362290 TI - Beta-amylases from various fungal strains. A review. AB - An overview presentation is made on the current global status of fungal beta3 amylases, their characteristics and applications in various industries. Among the few available report on beta-amylase producing fungal strains, many showed a preference for a cultivation temperature of 28 degrees C, acidic pH and soluble starch as an inducer of enzyme synthesis. In some fungal strains, alpha-amylase and alpha-glucosidases were found to be present as major contaminating enzymes. Although the existence of a few starch digesting and raw starch adsorbing fungal strains were reported, detailed study on molecular biology of corresponding fungal genes was not available. PMID- 15362291 TI - Evolution in action: molecular evidence for recent emergence of secotioid genera Endxoptychum, Gyrophragmium and Longula from Agaricus ancestors. AB - Phylogenetic analyses of 29 Agaricus species and one representative from genera Endoptychum, Gyrophragmium and Longula each were conducted based on sequence data of the entire internal transcribed spacers and partial large subunit of ribosomal DNA. The Agaricus species formed several distinct clades both confirming and challenging previous morphological sections in several cases. Endoptychum depressum, Gyrophragmium dunalii and Longula texensis were nested among species of the genus Agaricus. This study provides evidence for independent emergence of these secotioid fungi from Agaricus ancestors, most likely from species placed in the section Arvenses as inferred from molecular data. PMID- 15362292 TI - Why it is time to review the role of private health insurance in Australia. AB - The role of private health insurance (PHI) within the Australian health-care system is urgently in need of comprehensive review. Two decades of universal health cover under Medicare have meant a change in the function of PHI, which is not reflected in policies to support PHI nor in the public debate around PHI. There is increasing evidence that the series of policy adjustments introduced to support PHI have served to undermine rather than promote the efficiency and equity of Australia's health care system. While support for PHI has been justified to 'take pressure off the public hospital system' and to facilitate choice of insurer and private provider', and the incentives have indeed increased PHI membership, this increase comes at a high cost relative to benefits achieved. The redirection of hospital admissions from the public to private hospitals is small, with a value considerably less than 25% of the cost of the policies. The Commonwealth share of the health care budget has increased and the relative contribution from private health insurance is lower in 2001-02, despite an increase in PHI membership to nearly 45% of the population, compared with the 30% coverage in 1998. The policies have largely directed subsidies to those on higher incomes who are more likely to take out PHI, and to private insurance companies, private hospitals and medical specialists. Ad hoc policy adjustments need to be replaced by a coherent policy towards PHI, one that recognises the fundamental change in its role and significance in the context of universal health coverage. PMID- 15362293 TI - Some demographic issues affecting private health insurance. AB - There will be significant changes in the demography of persons with Private Health Insurance (PHI). Two methods of projecting PHI coverage are discussed in this paper. The first assumes the only factors affecting PHI coverage are demographic change and mortality and facilitates comparisons between actual and projected PHI coverage. The second projects the percentage of the population insured in each five year age cohort, and makes allowance for changes in PHI coverage due to all factors. Demographic change will increase Registered Health Benefit Organization (RHBO) premiums by 1.7% per annum. The role of these projections in analysing the effect of future premium increases on PHI retention rates is also discussed. PMID- 15362294 TI - Impact of private health insurance incentives on obstetric outcomes in NSW hospitals. AB - The purpose of this study is to analyse the impact of changes in Australian private health insurance coverage as seen in NSW public and private hospital birth profies, and identify trends in obstetric outcomes from 1997-2001. NSW Midwives Data Collection unit record data is analysed for women who gave birth to a live singleton baby of term gestation (> or = 37 weeks) and cephalic presentation in NSW hospitals during 1997- 2001. Use of private hospitals for childbirth has increased in conjunction with increases in private health insurance coverage. Although some obstetric interventions have increased for both public and private hospitals over time, clinical factors do not explain the large differences in birth interventions and outcomes between NSW public and private hospitals. Incentives to increase private health insurance coverage appear to be having a negative impact on childbirth, in terms of higher birth intervention and operative birth rates in NSW private hospitals. PMID- 15362295 TI - Generic drugs: international trends and policy developments in Australia. AB - Public and private third-party payers in many countries encourage or mandate the use of generic drugs. This article examines the development of generics policy in Australia, against the background of a description of international trends in this area, and related experiences of reference pricing programs. The Australian generics market remains underdeveloped due to a historical legacy of small Pharmaceutical Benefits Scheme price differentials between originator brands and generics. It is argued that policy measures open to the Australian government can be conceived as clustering around two different approaches: incremental changes within the existing regulatory framework, or a shift towards a high volume/low price role of generics which would speed up the delivery of substantial cost savings, and could provide enhanced scope for the financing of new, patented drugs. PMID- 15362296 TI - Promoting the health of older Australians: program options, priorities and research. AB - Relatively little emphasis has been placed on identifying health promotion research and program priorities for the older age group. A one-day conference culminating in an interactive session was organised to engage health service professionals in a process to identify such priorities in Western Australia. Physical activity social isolation, mental health and medications were deemed issues warranting more attention by both health promotion research and health promotion intervention programs. Additional consultation with representatives of the target population is recommended to further refine the priorities. PMID- 15362297 TI - 'Closing the loop': the role of health care leaders in integrating research and practice. AB - The study reported here examines what health care leaders do to implement the routine integration of information into clinical practice decision making. Clinical leaders of a major Australian tertiary teaching hospital were interviewed. The study found that there is wide variation in the extent to which information is routinely used to inform practice. Those leaders who successfully integrated information used some common strategies. The ownership of information may be the single most powerful factor in clinical leaders' ability to encourage its use. A lack of trust, inappropriate systems and a fear of unfavourable consequences were identified as issues. The findings have implications for policy makers, hospital administrators and clinical leaders and question the effectiveness of centralised approaches to driving clinical change. PMID- 15362298 TI - Type of housing predicts rate of readmission to hospital but not length of stay in people with schizophrenia on the Gold Coast in Queensland. AB - Accommodation is considered to be important by institutions interested in mental health care both in Australia and internationally Some authorities assert that no component of a community mental health system is more important than decent affordable housing. Unfortunately there has been little research in Australia into the consequences of discharging people with a primary diagnosis of schizophrenia to different types of accommodation. This paper uses archival data to investigate the outcomes for people with schizophrenia discharged to two types of accommodation. The types of accommodation chosen are the persons own home and for-profit boarding house. These two were chosen because the literature suggests that they are respectively the most and least desirable types of accommodation. Results suggest that people with schizophrenia who were discharged to boarding houses are significantly more likely to be readmitted to the psychiatric unit of Gold Coast Hospital, although their length of stay in hospital is not significantly different. PMID- 15362299 TI - Patient satisfaction: the Australian patient perspective. AB - The literature reveals little Australian academic study of the phenomenon of patient satisfaction and identifies several problems in current research practice. A theoretical discussion about the phenomenon of patient satisfaction' is for the most part absent, the rigour in the methods applied is often dubious, a definition of patient satisfaction is not agreed and the patient experience is often not the focus of research. To address some of these issues inductive research was conducted with Australian patients to explore what they considered important for patient satisfaction to exist. A series of 52 interviews were conducted with twenty elective surgery patients in an Australian teaching hospital Patients were interviewed on admission to hospital, within one week of discharge from hospital and between six and eight weeks after discharge. Research with patients identified 16 themes that were important to make a patients hospital stay satisfactory. Qualitative data have provided a foundation to better understand what patient satisfaction' means in its everyday use. Such an approach is faithful to the concerns and priorities of the patients who are the users of health care services. PMID- 15362300 TI - Management competencies: intrinsic or acquired? What competencies are required to move into speech pathology management and beyond? AB - Speech pathology managers frequently move into careers beyond their clinical discipline. As practicing speech pathology managers and students of business leadership, we were curious about the nature of career transitions out of speech pathology management. We conducted an exploratory descriptive study investigating the perceived competencies that facilitate such career transitions and when further education is required to effectively equip one for such transition. The perceived skills related to a speech pathology background are identified along with the gaps in competence for moving into general management positions. Career management practices that facilitate this type of career transition are provided as recommendations for career planning. PMID- 15362301 TI - Data, circumstance and politics: reflections on regional mental health planning. AB - We use our experience as consultants to a regional mental health planning project in South Australia to describe three practical aspects of regional health planning. First, we systematically summarised various data on socio-demographic indicators, health status and health service use along with qualitative opinion about needs and services from consultations with over 200 stakeholders. In addition to these data, we found that attention to two other aspects of planning, circumstance and politics, were of critical importance, particularly if the plan was to be implemented and as a way of turning thinking into action. PMID- 15362302 TI - Using nursing information systems to enhance quality service across multiple service providers. AB - This study explored the key requirements identified by stakeholders for the integration of Nursing Information Systems (NIS) in three public hospitals in the southern suburbs of Adelaide. The study used a qualitative approach of semi structured interviews, focus groups, site visits to the hospitals and review of relevant literature to ascertain what participants saw as the necessary ingredients to create regional NIS. Study findings showed that the current NIS are not sustainable in terms of staffing levels, physical resources or the capacity of the two currently-used computer products to interface with newer generation products. A critical issue for adopting a regional model is the willingness of local sites to revise their current internal structures and functions. This restructuring towards a regionalised NIS would improve overall communication, education, security efficiency and sustainability. PMID- 15362303 TI - The impact of caring on caregivers' mental health: a review of the literature. AB - This paper provides a brief overview of the literature on the impact of caring on the mental health of informal caregivers in the areas of aged care, disability and mental health. Factors discussed that may impact on caregivers' mental health include the relationship between the caregiver and care recipient, the nature of the care recipient's disability and the stage of the caregiving process. Several moderators of the impact of caring on mental health are described: including socio-economic factors, social support and coping strategies. This information provides a context within which to examine the type of interventions appropriate to assist caregivers in various situations. PMID- 15362304 TI - Rethinking midwifery refresher programs as a recruitment strategy. AB - This paper presents a case study on an innovative Midwifery Refresher Program in the context of current midwifery workforce issues. The refresher program was developed specifically as a recruitment strategy to address a staffing crisis at the Mater Misericordiae Mothers' Hospital, a busy tertiary maternity hospital in Brisbane, Australia. Features of the program that contributed to its success include collaboration with an industry partner, high levels of clinical support for participants, flexibility for women with family responsibilities and low financial costs for all stakeholders. This type of recruitment strategy may be useful for health care services experiencing staff shortages. PMID- 15362305 TI - Improving human resource management in Chinese healthcare: identifying the obstacles to change. AB - Health sector reform in China has led to increasing responsibility for hospital managers in the management of staff; but constraints continue. New personnel reforms offer new opportunities but face a number of difficulties. Drawing on research in Chinese hospitals in 1997 this paper identifies two major obstacles to improved human resource management: wage policy and lack of control by local managers over staffing. PMID- 15362306 TI - A case study in mainstreaming flexible learning in health--perspectives from the bush. AB - Our mission is to inculcate an ethos of online learning and communicating, moving it from the margins to mainstream. A skilled health workforce is crucial to better practices. Face-to-face teaching still dominates learning across NSW Health, despite remarkable developments worldwide in using technology for learning. Health is slow to embrace this renaissance due partly to lack of exposure among educators and learners, and the fact of course that learning is not its core business. The three staff comprising New England Area Health Service (NEAHS) Learning Services extensively researched NEAHS staff attitudes to fexible learning (2001) and Information Technology skills (2003). Amalgamating these data, with that from the first ever across-NSW Health online course we ran (2002) determined the appropriateness of our decision to instigate an external web-based discussion facility previously not available in health, for supporting learning and for communicating within NEAHS and indeed across Area Health Services (AHS). PMID- 15362307 TI - Sport is more important to health than most realise. PMID- 15362308 TI - Testing with confidence: the use (and misuse) of confidence intervals in biomedical research. PMID- 15362309 TI - Communicating the uncertainty in research findings: confidence intervals. AB - Confidence intervals (CIs) are an effective means of quantifying the uncertainty inherent in study results. Alongside considerations of possible sources of bias in the study design, CIs describe the extent to which study results are applicable in general, beyond the participants involved in the study. We discuss CI use and interpretation for some commonly encountered study designs. We highlight a close relation between CIs and p-values, such that presentation of 95% CIs can make it redundant to state whether a corresponding p-value is less than 0.05. We encourage researchers to use CIs to present their research findings, rather than relying on p-values alone. PMID- 15362310 TI - Training volume and strength and power development. AB - The purpose of this study was to determine if three training loads equated by volume differed in terms of the temporal, kinematic and kinetic characteristics of each set. Twelve experienced weightlifters (30.2+/-10.6 years old and 75.8+/ 13.0 kg) performed three sets (6 x 30% 1RM, 3 x 60% 1RM and 2 x 90% 1RM) of ballistic squats on an instrumented supine squat machine. Repeated measures ANOVA and Tukey HSD post hoc comparisons were used to distinguish significant differences between the three training loads on a variety of temporal, kinematic and kinetic variables. Significantly (p< 0.05) greater total time under tension during the eccentric (41-53%) and concentric phases (27-31%) was observed for the 30% 1RM condition compared to the other two conditions. Similarly, the lighter loading intensity resulted in significantly greater total eccentric (9-19%) and concentric (14-24%) force output compared to the other two conditions. Greater total power output was associated with the 30% 1RM condition for both the eccentric (25-48%) and concentric (40-69%) phases. Greater total work (9-24%) was also associated with the 30% 1RM condition. The 60% 1RM condition produced significantly greater total work, force and power compared to the 90% 1RM condition. However, greater concentric impulse (29-42%) was associated with the 90% 1RM condition. It is suggested that strength and power research needs to adopt a set kinematic and kinetic analysis approach within the research designs so that a better understanding of the nature of the neuromuscular adaptations elicited by different loading parameters is achieved. PMID- 15362311 TI - Impact energy attentuation of protective football headgear against a yielding surface. AB - There is no standard test to assess the ability of soft headgear to prevent the likelihood of head injuries in the Australian football codes. Therefore the impact energy attenuation of protective football headgear was assessed using a yielding artificial headform and impact surface to characterise the impact scenario. Seven soft football headgear of varying mass, materials and thickness were dropped from a height to generate impact energy of 56 J. The headgear were tested for a number of different impact locations. Accelerometers within the headform monitored the impact mechanics. A Head Injury Criterion (HIC) of 1000 was selected as the injury threshold. Reliability data, collected one week apart, revealed that the results were highly reproducible. Although several of the headgear generated HIC values about 1000 for the various impact locations, only one of the headgear, that with the greatest thickness (15 mm) generated a HIC score below 1000 for impacts to the side and front of the headgear. There was a significant correlation between headgear thickness and HIC scores. It was concluded that all but one of the commercially available soft football headgear tested provided inadequate impact energy attenuation due to the limited amount of padding. It is postulated that the padding material of the headgear would need to be at least 15 mm thick to offer adequate protection. PMID- 15362312 TI - Physical demands of elite Rugby League referees, part two: heart rate responses and implications for training and fitness testing. AB - The purpose of this study was to assess whether any characteristic patterns of heart rate (HR) responses could be identified in National Rugby League (NRL) referees (n= 6) during matches played in the 2001 season. The data have been plotted and discussed, in order that exercise program planning practitioners may gain improved understandings of the physiological requirements for referees. Some specific training suggestions have also been made. METHODS: The HR was recorded every five seconds throughout six competition NRL matches; using a heart rate monitor with a built-in memory. RESULTS: The specific magnitudes of referees' HR mean values varied between individuals, possibly due to specific game intensities, referee fitness, and age. All referees however exhibited similar HR response patterns; characterised by frequent (13-20 per match) large transient upward and downward shifts (>20 beats.minute(-1)). Periods of elevated HR extended for between five sec and eight min at a time, and were further characterised as a typical cyclic wave of HR elevation and recovery (ranging from 99.2+/-12.4 beats.minute(-1) to 176.5+/-11.8 beats.minute(-1) [mean +/-95% CI]), with a work to rest ratio of 2:1. Steady state HR was not achieved at any time during any match. CONCLUSIONS: The findings of the present study, taken together with a recent motion analysis, indicate that Rugby League refereeing is a highly intermittent, variable intensity activity. Significant anaerobic contribution to performance appears likely. It is suggested that training and fitness assessment of athletes should reflect their specific demands; some specific recommendations have therefore been provided. PMID- 15362313 TI - Self-consciousness and trait anxiety as predictors of choking in sport. AB - The present study was designed to examine dispositional self-consciousness and trait anxiety as predictors of choking in sport. Sixty-six basketball players completed the Self-Consciousness Scale and the Sport Anxiety Scale prior to completing 20 free throws in low-pressure and high-pressure conditions. A manipulation check showed that participants experienced significantly higher levels of state anxiety in the high-pressure condition. A series of hierarchical multiple regression analyses supported the hypothesis that self-conscious athletes were more susceptible to choking under pressure. The best predictors of choking were private self-consciousness and somatic trait anxiety that together accounted for 35% of the explained variance. We discuss a number of possible explanations regarding the discrepancy between the present results and previous studies mainly relating to task characteristics, skill level of participants and manipulations of pressure. PMID- 15362314 TI - The effect of interstate travel on sleep patterns of elite Australian Rules footballers. AB - The purpose of the current study was to examine the effect of interstate air travel on the quality and quantity of sleep in elite Australian Rules football players. Ten elite male athletes, who were members of a Western Australian-based Australian Football League (AFL) team, participated in the study. Sleep pattern was assessed by measuring sleep duration (SLD), sleep efficiency (SE), number of wakings (NW) and total wake time after sleep onset (WT) using a wrist-worn actigraph. Subjective sleep quality (SQ) was assessed using a scale of sleep rating. Throughout the 2002 AFL season, measurements were obtained on the night before (N1), the night of (N2) and the night after (N3) home and away games. Baseline measurements were obtained from five consecutive non-game nights. Compared to baseline, SLD on N1 was increased when home and away (by 51 and 105 mins respectively, p<0.05), while all other measures of sleep pattern were unchanged. On N2, SLD was decreased to a similar degree whether home or away (by 68 and 64 mins respectively, p<0.05), while all other measures of sleep pattern were unchanged. By N3 all measures of sleep pattern had returned to baseline values. Relative to baseline, perception of SQ was worst on N2 of a home game. This study has shown that interstate travel by elite AFL players has no adverse effects on sleep pattern on the night before a game. PMID- 15362315 TI - Enhancing specificity in proxy-design for the assessment of bioenergetics. AB - The purpose of this study was to examine the hypothesis that improved prediction of bioenergetics may be achieved when proxies are designed to simulate closely gold standard laboratory protocols. To accomplish this, a modified 'square' variation (SST) of the classical 20m Multistage Shuttle Run Test (MST) was designed aiming to reduce the stopping, turning and side-stepping manoeuvres. Within two weeks, 50 male volunteers (age 21.5+/-1.6, BMI 24.4+/-2.2) randomly underwent three maximal oxygen uptake (VO2max) assessments using a treadmill test (TT), the SST and MST. To assess SST reproducibility, 10 randomly-selected subjects performed the test twice. Validity results showed that mean predicted VO2max from SST was not significantly different compared to TT VO2max (p>0.05). In contrast, the equivalent value from MST was significantly higher (p<0.001) than TT. Furthermore, TT VO2max correlated with SST and MST at r=0.88 (p<0.001) and r=0.61 (p<0.05), respectively. The '95% limits of agreement' analysis (LIM(AG)) for SST and MST indicated a range of error equal to -0.5+/-5.4 and 8.1+/-8.0 (ml.kg(-1).min(-1)) with a coefficient of variation of +/-6 and +/ 8.2%, respectively. Test-retest results for SST revealed no mean difference in VO2max (p>0.05) and a correlation coefficient of r=0.98 (p<0.001), while LIM(AG) demonstrated a range of error equal to -0.2+/-2.6 (ml.kg(-1).min(-1)) with a coefficient of variation of +/-5.6%. It is concluded that, compared to MST, the SST had a higher agreement with TT. The latter may well be explained by the closer simulation in bioenergetics between the two protocols (ie, the continuous nature of SST provides a closer proxy of TT). PMID- 15362316 TI - Test-retest reliability of four physical activity measures used in population surveys. AB - Accurate monitoring of prevalence and trends in population levels of physical activity (PA) is a fundamental public health need. Test-retest reliability (repeatability) was assessed in population samples for four self-report PA measures: the Active Australia survey (AA, N=356), the short International Physical Activity Questionnaire (IPAQ, N=104), the physical activity items in the Behavioral Risk Factor Surveillance System (BRFSS, N=127) and in the Australian National Health Survey (NHS, N=122). Percent agreement and Kappa statistics were used to assess reliability of classification of activity status as 'active', 'insufficiently active' or 'sedentary'. Intraclass correlations (ICCs) were used to assess agreement on minutes of activity reported for each item of each survey and for total minutes. Percent agreement scores for activity status were very good on all four instruments, ranging from 60% for the NHS to 79% for the IPAQ. Corresponding Kappa statistics ranged from 0.40 (NHS) to 0.52 (AA). For individual items, ICCs were highest for walking (0.45 to 0.78) and vigorous activity (0.22 to 0.64) and lowest for the moderate questions (0.16 to 0.44). All four measures provide acceptable levels of test-retest reliability for assessing both activity status and sedentariness, and moderate reliability for assessing total minutes of activity. PMID- 15362317 TI - Do community football players wear allocated protective equipment? Descriptive results from a randomised controlled trial. AB - Before protective equipment can be adopted as an effective sports safety intervention, its protective effects in reducing the incidence and severity of injury need to be demonstrated, Importantly, it also needs to be well accepted by the players. The Australian Football Injury Prevention Project (AFIPP) was a large scale community-based randomised controlled trial to assess the effectiveness of headgear and mouthguards in preventing head/neck/dental injuries in Australian Football. A subcomponent of this study was to assess the extent to which community football players complied with the requirement to wear protective headgear and/or mouthguards, as this equipment is not compulsory in this sport. Three hundred and one community football players from 23 teams were randomly allocated to one of three protective equipment intervention arms or one control arm. Protective equipment usage was measured by a primary data collector at each training and game session during the 2001 playing season. Mouthguard use was higher than headgear use, with the highest usage for both being measured during games rather than training. Although many players use mouthguards, particularly in games, most do not wear headgear. Given the low adoption of headgear, other strategies to prevent head injuries need further investigation. PMID- 15362318 TI - Penalty shot importance, success and game context in international water polo. AB - To establish the incidence, timing and quantitative importance of penalty shots in water polo and to test whether or not penalty shot success would vary with the context (closeness, quarter, criticality) of the game, official records from six major international tournaments (n= 296 games) were analysed. Across all tournaments, penalties (n= 206) were awarded (1-3 per game) in 51% of games with no difference in frequency between game quarters. Penalty goals (n= 165) comprised only 3.7% of all goals scored, whereas the outcome of penalties (goal/no goal) within each game affected the final outcome (win/loss/tie) of 20% of games. The success rate of penalty shots (80.1%) was not significantly different between games classed as either close or non-close, by a mathematical expression of the running average goal difference up until the time of the penalty, and by the absolute difference of the score at the time of the penalty. Nor was this success rate significantly different between game quarters (72.7, 83.0, 81.5, and 81.8%), or between games classified by their criticality to final tournament placing (80.0, 79.5, and 80.6%, from highest to lowest). Thus, during international water polo, penalties contribute only modestly to game outcome, and penalty shot success is not significantly related to the closeness, quarter, or criticality of the game being played. PMID- 15362319 TI - Sport safety policies and practices in two rural Victorian communities. AB - Australian football and netball are the predominant sports played in rural Victoria, Australia. This exploratory study is the first to report the sport safety policies and practices adopted by junior Australian football and netball clubs in small rural communities. Eleven informants from four clubs completed a semi-structured interview and survey. Whilst the clubs performed a range of injury prevention activities, they did not have formal sports safety policies. Generally, netball informants reported fewer safety practices than football informants. Crucial factors influencing safety policies and practices were the reliance on volunteers and a lack of senior players. Barriers towards the adoption of safety policies and practices appeared to be related to rural population declines, a lack of qualified people and attitudes to injury in rural areas. Future research needs to identify how widespread this lack of sport safety policies and practices is across rural Australia and to identify strategies to overcome barriers to implementing them. PMID- 15362320 TI - The relationship between fast bowling workload and injury in first-class cricketers: a pilot study. AB - This study examined the relationship between the bowling workload of first-class fast bowlers and injury with the aim of identifying a "safe" fast bowling workload threshold. Twelve male fast bowlers (mean age 25 years) from an Australian state cricket squad were observed for the 1999--2000 cricket season. Workload was quantified by examining fixture scorecards and conducting surveillance at training sessions. Injury data were obtained from Cricket Australia's Injury Surveillance System. The seasonal incidence of injury was high with seven bowlers sustaining nine injuries. Whilst injured bowlers did not tend to bowl a greater number of deliveries on the day of injury, a significant increase in deliveries per session was observed in the 8-21 days prior to the date of injury (mean= 77) as compared with the rest of the season (mean= 60, p< 0.02). Bowlers with a weekly bowling workload above the mean of 203 deliveries were at an increased risk of injury (Risk Ratio (RR)= 6.0, 95% confidence interval (CI) 1.00-35.91). Those bowlers who bowled in five or more sessions in any 7-day period were also at an increased risk of injury (RR= 4.5, 95% CI 1.02 to 20.12). A consistent relationship between high bowling workload and injury was observed. The risk of injury was much higher for those bowlers with a sessional, weekly and monthly bowling workload above the group mean, especially when this high workload was consistent and sustained. PMID- 15362321 TI - CD94 expression and natural killer cell activity after acute exercise. AB - This study examined the effects of acute exercise on natural killer (NK) cell numbers, their expression of CD94 and cytotoxic capacity in triathletes over a 10 week training period. Nine highly trained male triathletes (age 25.9+/-4.1 yrs, VO2max 5.14+/-0.33 L.min(-1)) attended the laboratory on weeks 0, 2, 5 and 10 for incremental submaximal and maximal cycle ergometry. Peripheral blood was analysed for white blood cell counts, lymphocyte phenotype and cytolytic activity (51Cr release from K562 cells). Maximum oxygen consumption increased from week 2 (5.14+/-0.33 L.min(-1)) to week 10 (5.28+/-0.32 L.min(-1)). Resting NK cell numbers and their expression of CD94 were not altered over the 10-week study period. Natural killer cells expressing CD94+ were not differentially recruited into the circulation and cytolytic activity of exercise-recruited NKs did not differ from those present at rest. There was longitudinal stability (over the 10 weeks of the study) in CD94 expression on NK cells, exercise recruitment of CD94+ NK cells and cytolytic capacity of NK cells. The distribution and functional activity of NK cells are not markedly influenced by 10 weeks of training in competitive triathletes. Natural killer cytotoxic activity after exercise reflects numbers of NK cells and not a changed activation state of these cells per se. PMID- 15362322 TI - The accuracy of MRI in predicting recovery and recurrence of acute grade one hamstring muscle strains within the same season in Australian Rules football players. AB - The purpose of this study was to use MRI to classify acute grade one hamstring muscle strains in Australian Rules footballers to determine if it was accurate in predicting the recovery time for each injury and also able to predict those that would recur within the same season. A prospective study was performed over five years at a professional Australian Football League club. Thirty-one acute grade one hamstring injuries underwent MRI examination within 24-72 hours following the injury. Each injury underwent the same rehabilitation programme. The rehabilitation interval (RI) was the time in days for the player to resume full team training. Fourteen (45%) of the injuries were normal on MRI. Seventeen (55%) were abnormal with a hyperintense T2 lesion on the axial fat suppressed views. The MRI negative group had a significantly faster RI (6.6 days) compared with the MRI positive group (20.2 days). Both the length and cross sectional area (CSA) of the MRI positive lesions were measured. The length of the lesion had a stronger correlation coefficient with the RI (0.84) than did the CSA (0.76). Six of the 17 MRI positive strains recurred with no correlation found between the lesion's length or CSA, or the RI. None of the 14 MRI negative injuries recurred. The study confirms that MRI can aid in the investigation of acute grade one hamstring muscle strains in predicting recovery time. However the size of the initial strain or the RI do not seem to be reliable indicators in predicting those strains that might recur. PMID- 15362323 TI - Severity of viral infection is promoted by hyperthermic pretreatment. AB - The aim of this study was to determine if prior heat shock and consequential heat shock protein (HSP) accumulation would inhibit viral infection. Confluent cultures of Madin-Darby bovine kidney cells were subjected to 30 mins of hyperthermia at 42 degrees C then allowed to recover at 37 micro C for six hrs. During the six hrs, HSP concentrations increased by 232% in heat-shocked cultures. Following the recovery period, six replicates of heat-shocked (HS) and six replicates of non-heat-shocked (CON) cultures were infected with Bovine Herpes virus 1. After 72 hours, severity of infection was assessed by counting formed viral plaques in cell cultures. CON cultures averaged 172.0+/-34.3 plaques per culture and HS cultures averaged 339.3+/-96.1 plaques per culture, representing a significant increase in infected cells (p=0.0044). These data provide evidence that physiological stressors related to extreme or exhaustive exercise, specifically hyperthermia, may increase risk of viral infection. PMID- 15362324 TI - The introduction of new technology. PMID- 15362325 TI - Delivery of a surgical clerkship program in a remote site: personal experiences and published evidence. PMID- 15362326 TI - Efficacy of orthotic immobilization of the unstable subaxial cervical spine of the elderly patient: investigation in a cadaver model. AB - OBJECTIVE: To assess the efficacy of soft, semirigid and hard cervical collars to immobilize the neck in a destabilized cadaver model. DESIGN: This is a laboratory experiment. SETTING: The anatomy research lab of McMaster University. PATIENTS: None. Fresh cadavers from elderly patients suffering terminal medical illness and free of cervical structural disease were studied. INTERVENTIONS: Destabilizing discoligamentous lesions of the neck were created in the cadavers. Radiographs were taken in maximum displacement in the prone, decubitus and side-bending positions, first unsupported and then with soft, semirigid and hard collars applied. Displacements in angulation and translation were measured from the radiographs. OUTCOME MEASURES: Radiographic displacement under gravity load. RESULTS: In all cases there was no effective limitation of pathological displacement, and in many cases displacement was increased after collar application. CONCLUSIONS: Cervical collars do not effectively support the unstable neck, and may be ineffective in preventing pathological displacements. PMID- 15362327 TI - Branched endovascular stent-graft for suprarenal aortic aneurysm: the future of aortic stent-grafting? AB - The use of a branched endovascular stent-graft to repair an aneurysm of the visceral aorta is described. The evolving role of branched endovascular stent grafts in the management of aortic aneurysms is discussed, and the literature reviewed. PMID- 15362328 TI - Can a new design of pneumatic compression device reduce variations in delivered therapy for the mechanical prophylaxis of thromboembolic disease after total hip arthroplasty? AB - BACKGROUND: Compression devices have been shown to prevent thromboembolic disease. However, the pressures generated may not be the same as the ones recommended by the manufacturer. The purpose of this study is to investigate a new sequential compression device with feedback to maintain optimal therapy, and to determine whether therapy is improved with this new device. PATIENTS AND METHOD: A series of 50 patients undergoing elective total hip arthroplasty at a major tertiary-care hospital with a special interest in joint replacement were enrolled prospectively. In addition to pharmacological prophylaxis for thromboembolic disease, all patients received compression from a modified device. Maximum pressures generated and the rate of pressure rise in each of the 3 compartments within the device sleeves were measured and the results compared with data from historical controls. RESULTS: We considered therapy to be ideal when in a particular compression cycle all chambers of both right and left sleeves reach within 10% of their target pressures at within 10% of their target pressure rise rates. The average patient received this ideal therapy 88% of the time that the new trial sequential compression device was operating. This represents a dramatic improvement over previous devices. CONCLUSIONS: The new device allows dramatically improved pressures within the device because of a feedback loop that allows dynamic control of each chamber's pressure. Improved consistency of delivery should make it easier to accurately assess the true benefits of mechanical prophylaxis with a sequential compression device. PMID- 15362329 TI - Improving cancer surgery in Ontario: recommendations from a strategic planning retreat. AB - INTRODUCTION: The Ministry of Health and Long-Term Care mandated a rapid and thorough change in the delivery of cancer services in Ontario to integrate ambulatory services offered by Cancer Care Ontario (CCO) with the inpatient services of affiliated hospitals. The CCO Surgical Oncology Program held a strategic planning retreat to establish the basis upon which to implement surgery specific changes. METHODS: Participants completed a pre-retreat survey. Based on survey results, the retreat was organized around 4 themes: role of the Surgical Oncology Program; knowledge transfer; funding for cancer surgery; and research priorities. These topics were discussed in small breakout groups and by the entire assembly. RESULTS: Retreat participants (n = 55) included hospital CEOs, vice-presidents of cancer services, surgeons from cancer centres and community hospitals, academic chairs of surgery, clinician researchers and managers from CCO. Responses to the pre-retreat survey (n = 38) and recommendations made by retreat participants showed strong support for the Surgical Oncology Program to take a leadership role in the development and monitoring of quality indicators, research related to cancer surgery and the creation of regional communities of practice. Funding mechanisms for cancer surgeons and hospitals performing cancer surgery were also highlighted. CONCLUSION: The Surgical Oncology Program used the results to develop a strategic plan that was approved by retreat participants and the board of the CCO. The program has embarked on a multifaceted approach to facilitate, monitor and report on the organization and delivery of cancer surgery in Ontario. PMID- 15362330 TI - Teaching cognitive skills improves learning in surgical skills courses: a blinded, prospective, randomized study. AB - OBJECTIVE: To investigate the teaching of cognitive skills within a technical skills course, we carried out a blinded, randomized prospective study. METHODS: Twenty-one junior residents (postgraduate years 1-3) from a single program at a surgical-skills training centre were randomized to 2 surgical skills courses teaching total knee arthroplasty. One course taught only technical skill and had more repetitions of the task (5 or 6). The other focused more on developing cognitive skills and had fewer task repetitions (3 or 4). All were tested with the Objective Structured Assessment of Technical Skill (OSATS) both before and after the course, as well as a pre- and postcourse error-detection exam and a postcourse exam with multiple-choice questions (MCQs) to test their cognitive skills. RESULTS: Both groups' technical skills as assessed by OSATS were equivalent, both pre- and postcourse. Taking their courses improved the technical skills of both groups (OSATS, p < 0.01) over their pre-course scores. Both groups demonstrated equivalent levels of knowledge on the MCQ exam, but the cognitive group scored better on the error-detection test (p = 0.02). CONCLUSIONS: Cognitive skills training enhances the ability to correctly execute a surgical skill. Furthermore, specific training and practice are required to develop procedural knowledge into appropriate cognitive skills. Surgeons need to be trained to judge the correctness of their actions. PMID- 15362331 TI - Slipped capital femoral epiphysis: an analysis of treatment outcome according to physeal stability. AB - INTRODUCTION: Slipped capital femoral epiphysis (SCFE) is a common hip disorder in adolescents. Treatment can be associated with serious complications, mainly avascular necrosis (AVN). The objective of this study was to evaluate treatment outcomes and complications based on physeal stability at presentation. METHODS: We reviewed retrospectively all children treated for SCFE from 1990 through 2000 at the Children's Hospital of Eastern Ontario, Ottawa. RESULTS: Of a total of 87 patients (56 male) identified, 73 had slips classed as stable and 14, unstable; 42 involved the left side, 34 the right, and 19 were bilateral. Avascular necrosis (AVN) developed in 1 patient with a stable slip (morbidity 1.4%) and in 3 with unstable slips (21.4%; p = 0.001). Manipulative anatomical reduction was done in 11 cases (78.5%), all unstable slips. All slips complicated by AVN had been classified according to the traditional system as acute-on-chronic. Complication rates differed significantly between unstable slips (36%) and stable (20%). No statistically significant association was found between AVN and slip severity, time to fixation or use of reduction. CONCLUSIONS: The treatment of stable slips exhibits a lower incidence of AVN. These results nevertheless suggest that gentle reduction of unstable slips is safe and unaccompanied by an increase in complications. PMID- 15362332 TI - Surgical images: musculoskeletal. Imaging in musculoskeletal trauma: the value of magnetic resonance imaging for traumatic pediatric hip dislocations. PMID- 15362333 TI - Surgical Education and Self-Assessment Program (SESAP). Foot ulcers in the diabetic patient. PMID- 15362334 TI - Radiology for the surgeon: soft-tissue case 55. Sacral thumbtack sinus following proctocolectomy. PMID- 15362335 TI - Hemolysis with rapid transfusion systems in the trauma setting. AB - OBJECTIVE: Rapid infusion system allows rapid infusion of resuscitation fluids at body temperature in trauma patients. Packed red blood cells are subjected to high external pneumatic pressure (up to 300 mm Hg) and rapid infusion rates through a 170-microm filter. This study was conducted to outline hemolysis that may occur in the setting of massive transfusion (> 10 units). METHODS AND MATERIALS: Measurements of various parameters were made before and after infusion of 17 units of outdated (38-82 d) packed red blood cells through a Level 1 Rapid Infuser, including lactate dehydrogenase (LDH), potassium, plasma hemoglobin, hematocrit and total hemoglobin. Hemolysis, expressed as a percentage, was calculated from these parameters. RESULTS: Hemolysis observed in this experiment ranged from near 0 to 0.05%. All the units had plasma potassium concentrations of 15 mmol/L or more. CONCLUSION: Transfusion of 17 units with the Level 1 Rapid Infuser did not cause a clinically significant amount of hemolysis. PMID- 15362336 TI - Scapholunate ligament injury: the natural history. PMID- 15362337 TI - Split-thickness skin-grafting the post-Clagett pleurostomy cavity. PMID- 15362338 TI - Surface replacement of the hip: a late revision. PMID- 15362339 TI - Resident training. PMID- 15362340 TI - Snowmobile trauma, alcohol and the law. PMID- 15362341 TI - Paleoradiology: current status and future challenges. PMID- 15362342 TI - History of paleoradiology: early published literature, 1896-1921. PMID- 15362343 TI - Did Ramesses II really have ankylosing spondylitis? A reappraisal. AB - OBJECTIVE: To challenge the diagnosis of ankylosing spondylitis in the mummy of Ramesses II that was suggested about 30 years ago and to propose a differential diagnosis for the changes that were detected in the mummy's spine and pelvis. METHODS: We read and interpreted both the published and unpublished radiographs of the mummy. RESULTS: Changes in the mummy's spine and pelvis included ossification of the anterior longitudinal ligament at the cervical spine level, ossified enthesopathy of both the right and left rectus femoris tendons as well as the right ischial tuberosity, a large osteophyte at both acetabula margins, no ankylosis of the cervical apophyseal joints and no ankylosis of the right sacroiliac joint. CONCLUSION: The radiologic evidence does not support the claim that Ramesses II had ankylosing spondylitis. Our radiologic reappraisal suggests instead the diagnosis of diffuse idiopathic skeletal hyperostosis. This new diagnosis needs to be validated by a computed tomographic scan of the mummy. PMID- 15362344 TI - Diagnostic paleoradiology of mummified tissue: interpretation and pitfalls. PMID- 15362345 TI - Paleoradiology in mummy studies: the Sulman mummy project. PMID- 15362346 TI - Multislice computed tomography of two 2000-year-old skeletons in a soil matrix from Angkor, Cambodia. PMID- 15362347 TI - Micro-computed tomography of a 500-year-old tooth: technical note. AB - OBJECTIVE: To determine whether micro-computed tomography (micro-CT) could be used to reconstruct ancient dental anatomy accurately and differentiate the enamel from the dentin, as well as to verify whether micro-CT could detect tooth disorders such as attrition or caries accurately. METHODS: Micro-CT imaging was performed, using a cone-beam micro-CT specimen scanner, on a 500-year-old human tooth found in a burial jar in the Cardomom Mountains in southwestern Cambodia. RESULTS: The occlusal surface of the tooth showed marked attrition, with the dentin extending close to the enamel layer on the crown. In addition to this, micro-CT images depicted calculus on the buccal surface and a cervical root caries lesion present on the distal surface. The sclerotic zone of the carious lesion (located deep in the destroyed dentin) and the dentin were effectively differentiated through excellent resolution and superior tissue contrast of the volume data set. Axial slices from apical to coronal show the carious lesion extending vertically along the dentin-enamel junction with an intact outer enamel surface. CONCLUSION: Micro-CT is a reproducible, nondestructive and highly accurate technique that can be successfully applied to the study of ancient teeth. PMID- 15362348 TI - Field paleoradiography of skeletal material from the early classic period of Copan, Honduras. AB - OBJECTIVE: To perform a radiographic evaluation of the skeletal remains of 2 elite individuals from the Early Classic Period of Copan, namely, K'inich Yax K'uk' Mo' (Hunal Burial 95-2), founder of the Early Classic Dynasty, and the primary female interment from the Motmot tomb (Burial 37-8). Our aims were to document pathologies and subsequent changes in long-bone density resulting from physiologic or functional adaptations, such as disuse atrophy. METHODS: Standardized anteroposterior and mediolateral radiographs were obtained using a portable veterinary x-ray unit and Polaroid film. Orthogonal radiographs were used to estimate the cross-sectional biomechanical properties of the long bones. RESULTS: In the case of Yax K'uk' Mo', it appears that there was trauma-induced disuse atrophy of the right forearm resulting in an accelerated loss of cortical bone in the right humerus relative to the left humerus. The restructuring of this individual's left shoulder girdle probably resulted in radical structural and functional changes. Although this fracture could have contributed to degenerative changes and paralysis, there was no evidence of disuse atrophy in the left upper limb. The gracile Motmot skeleton showed no signs of osteoporosis, osteopenia or disuse atrophy. CONCLUSIONS: The use of radiography in the assessment of the skeletal material described here vastly increased our ability to document the pathologic processes that affected these centuries-old individuals during their lifetimes. In addition, the use of radiographs allowed us to expand our understanding of how these processes may have affected biomechanics and, subsequently, long-bone geometric properties. PMID- 15362349 TI - The application of radiography to field studies in physical anthropology. PMID- 15362350 TI - Dental paleoradiology: applications in paleoanthropology and paleopathology. AB - OBJECTIVE: To study the emergent role of computed tomography (CT) in the evaluation of ancient human dental remains. METHODS: We conducted a paleoradiologic study of the skeletal remains of 4 individuals dating from the Pliocene to the Medieval Period. The specialized software used permitted comprehensive analyses of the bone structures of the dental remains and the performing of three-dimensional stereolithographic reconstructions. RESULTS: Paleoradiology helps demonstrate the detailed anatomy of fossil specimens, which permitted us to assign 1 of the specimens that we studied to the genus Homo, and not Australopithecus. Tooth and mandibular disease were identified in 2 specimens. CONCLUSION: CT is an essential noninvasive and nondestructive tool for the evaluation of human dental remains. PMID- 15362351 TI - Three-dimensional computer imaging of hominid fossils: a new step in human evolution studies. PMID- 15362352 TI - Three-dimensional digital imaging of the partial Australopithecus africanus endocranium MLD 37/38. AB - OBJECTIVE: MLD 37/38 is one of a handful of Australopithecus africanus specimens. The partial cranium (about 2.5 million years old) is filled with stone matrix. The frontal part of the endocranium and most of the face are missing. Based on medical imaging techniques (computed tomography), we aimed to create virtual 3 dimensional reconstructions separately for the ectocranium and endocranium, reconstruct missing parts based on intraspecific comparison, estimate the endocranial capacity and describe endocranial morphological details that were physically inaccessible before. METHODS: We mapped fossilized bone as distinct from stone matrix for every slice of the computed tomographic image, creating virtual endocasts. The missing endocranial parts were reconstructed, using thin plate spline warping, from a reference specimen (STS 5, a complete A. africanus cranium). In contrast with previous studies, a large number of landmarks (8 anatomical and 455 semi-landmarks) were used for this reproducible reconstruction. RESULTS: The endocranial capacity of the original partial cranium is 382 cm3, and the estimated volume for the reconstructed braincase is 440 cm3. On the left side of the endocast, signs of the meningeal vessel system could be detected. Transverse and sigmoid venous sinuses are present in MLD 37/38, whereas an occipital-marginal sinus system could not be found. CONCLUSIONS: The reconstructed endocranial capacity of MLD 37/38 lies within the range of other A. africanus specimens (428-515 cm3). In its dimensions, the endocast is similar to that of STS 5. The patterning of detected meningeal arterial branches resembles that of modern humans. The absence of an enlarged occipital-marginal sinus system is typical of A. africanus. PMID- 15362353 TI - The ethics of self. AB - This article attempts to elucidate the ethical meaning behind the words 'the ethics of self' and 'an ethical self', particularly in the light of Noddings' 'ethical self', in order to show the relevance of these terms to the practice of ethical caring. It examines the relationship that Noddings believes exists between one's actual self and the vision one has of one's ideal self. I attempt to draw out a meaning derived from the texts in which this concept has been captured, while at the same time juxtaposing the concept 'ethic of care' from my own research, together with the meaning given by philosophers Peta Bowden, Iris Murdoch and Simone Weil, who have written on the subject, albeit indirectly in some instances. A participant in a colleague's research on care and patient satisfaction used similar expressions, and gave the impetus for this article. PMID- 15362354 TI - A qualitative analysis of ethical problems experienced by physicians and nurses in intensive care units in Turkey. AB - In this qualitative study, we aimed to identify and compare the ethical problems perceived by physicians and nurses in intensive care units at Baskent University hospitals in Turkey. A total of 21 physicians and 22 nurses were asked to describe ethical problems that they frequently encounter in their practice. The data were analyzed using an interactive model. The core problem for both physicians and nurses was end-of-life decisions (first level). In this category, physicians were most frequently concerned with euthanasia while nurses were more concerned with do-not-resuscitate orders (second level). At the third level, we saw that almost all of the participants' responses related to negative perceptions about euthanasia. Communication and hierarchical problems were the second most reported main category. Nurses were more likely to cite problems with hierarchy than physicians. At the third level, a large percentage of nurses described communication problems with authority and hierarchical problems with physicians. In the same category, physicians were most often concerned with communication problems with patients' relatives. The ethical problems were reported at different frequencies by physicians and nurses. We asked the participants about ethical decision-making styles. The results show that nurses and physicians do not follow a systematic pattern of ethical decision making. PMID- 15362355 TI - Relational ethics and genetic counseling. AB - Genetic counseling is viewed as a therapeutic interrelationship between genetic counselors and their clients. In a previous relational ethics research project, various themes were identified as key components of relational ethics practice grounded in everyday health situations. In this article the relational ethics approach is further explored in the context of genetic counseling to enhance our understanding of how the counselor-client relationship is contextually developed and maintained. Qualitative interviews were conducted with six adult clients undergoing genetic counseling for predictive testing. Engagement, dialogue and presence were revealed as relevant to genetic counselor-client relationships. A relational ethics approach in genetic counseling challenges the concept of nondirectiveness and may enhance the outcome of counseling for both counselor and client. PMID- 15362356 TI - Patient rights and law: tobacco smoking in psychiatric wards and the Israeli Prevention of Smoking Act. AB - In August 2001, the Israeli Ministry of Health issued its Limitation of Smoking in Public Places Order, categorically forbidding smoking in hospitals. This forced the mental health system to cope with the issue of smoking inside psychiatric hospitals. The main problem was smoking by compulsorily hospitalized psychiatric patients in closed wards. An attempt by a psychiatric hospital to implement the tobacco smoking restraint instruction by banning the sale of cigarettes inside the hospital led to the development of a black market and cases of patient exploitation in return for cigarettes. This article surveys the literature dealing with smoking among psychiatric patients, the role of smoking in patients and the moral dilemmas of taking steps to prevent smoking in psychiatric hospitals. It addresses the need for public discussion on professional caregivers' dilemmas between their commitment to uphold the law and their duty to act as advocates for their patients' rights and welfare. PMID- 15362357 TI - Important but neglected ethical and cultural considerations in the fight against HIV/AIDS in Malawi. AB - Southern African countries have the highest HIV infection rates in the world. In most of the countries in the region, the rate among adults is at least 10%. The fight against HIV/ AIDS has mostly been inadequate owing to the lack of proper consideration of ethical and cultural issues. In this article, the authors discuss the ethical and cultural dilemmas concerning HIV/AIDS, with Malawi as a case in point. It is argued that increasing financial resources alone, as exemplified by the Global Fund to Fight AIDS, Tuberculosis and Malaria initiative, without proper attention to ethical issues, morals and appropriate legal obligations, are unlikely to reduce the spread of HIV in southern Africa. PMID- 15362358 TI - Nursing and midwifery malpractice in Turkey based on the Higher Health Council records. AB - Medical malpractice has attracted the attention of people and the media all over the world. In Turkey, malpractice cases are tried according to both criminal and civil law. Nurses and midwives in Turkey fulfill important duties in the distribution of health services. The aim of this study was to reveal the legal procedures followed in malpractice allegations and malpractice lawsuits in which nurses and midwives were named as defendants. We reviewed 59 nursing and midwifery lawsuits reported to the Higher Health Council between 1993 and 1998. Health professionals were held liable in 59% of the lawsuits. Midwives had the highest percentage of malpractice liability (52%), followed by physicians (29%) and nurses (19%). To reduce potential liability, nurse education should be improved, a nursing malpractice law should be enacted, and instructions for nursing procedures should be formulated. PMID- 15362359 TI - Truth-telling in clinical practice and the arguments for and against: a review of the literature. AB - In general, most, but not necessarily all, patients want truthfulness about their health. Available evidence indicates that truth-telling practices and preferences are, to an extent, a cultural artefact. It is the case that practices among nurses and doctors have moved towards more honest and truthful disclosure to their patients. It is interesting that arguments both for and against truth telling are established in terms of autonomy and physical and psychological harm. In the literature reviewed here, there is also the view that truth-telling is essential because it is an intrinsic good, while it is argued against on the grounds of the uncertainty principle. Based on this review, it is recommended that practitioners ought to ask patients and patients' families what informational requirements are preferred, and research should continue into truth telling in clinical practice, particularly to discover its very nature as a cultural artefact, and the other conditions and contexts in which truth-telling may not be preferred. PMID- 15362360 TI - A Chinese perspective on autonomy. PMID- 15362361 TI - Country profile: Afghanistan. PMID- 15362362 TI - Performance characteristics of eight estradiol immunoassays. AB - Measurement of estradiol is useful in assisted reproduction, evaluation of infertility, menopause, and male feminization. The analytic performance of 8 estradiol immunoassays was evaluated. The imprecision and accuracy of the Access, ADVIA Centaur, ARCHITECT i2000, AutoDELFIA, Elecsys 2010, IMMULITE 2000, and Vitros ECi estradiol assays (see text for proprietary information) were evaluated by using an isotope dilution-gas chromatography-mass spectrometry (ID-GC-MS) reference method. The coefficient of variation (CV) ranged from 6.9% on the Elecsys 2010 to 42.6% on the ADVIA Centaur at an estradiol concentration of 18 pg/mL (66 pmol/L), with the ARCHITECT i2000 assay in development and the Vitros ECi having a CV below 10% at this estradiol concentration. Agreement between the automated assays and ID-GC-MS was variable, with slopes ranging from 0.87 to 1.20. The Access, ARCHITECT i2000 in development, and the IMMULITE 2000 were the most accurate, with slopes of 0.99, 0.98, and 1.03, respectively. These findings indicate that the ARCHITECT i2000 estradiol assay in development had the best precision and accuracy of the assays evaluated for measurement of serum estradiol concentrations. PMID- 15362363 TI - Race and the linkage between serum prostate-specific antigen and prostate cancer: a study of American veterans. AB - Many aspects of prostate cancer differ between black men and white men, including incidence, stage, grade, sensitivities and specificities of serum prostate specific antigen (PSA) levels, and survival. In general, the level of serum PSA reflects the mass of the prostate and the amount of tumor present, but the question to consider is whether this relationship is the same for blacks as for whites. If it is the same, then the ways we use serum PSA to screen, stage, and follow up white men with cancer should work equally for black men. If it is not, then we need alternative strategies for using serum PSA levels in blacks. I used regression analysis to study how the serum PSA level depends on prostate mass and the amount of tumor in 194 American veterans, including 87 black men. I found that black men produced higher levels of serum PSA for any given amount of tumor compared with whites, and I demonstrated that this difference can significantly affect the assessment of risk for outcomes in blacks. PMID- 15362364 TI - Acute myeloid leukemia with t(6;9)(p23;q34) is associated with dysplasia and a high frequency of flt3 gene mutations. AB - We report 12 cases of t(6;9)(p23;q34)-positive acute myeloid leukemia (AML), all classified using the criteria of the World Health Organization classification. There were 10 women and 2 men with a median age of 51 years (range, 20-76 years). Dysplasia was present in all cases (9 previously untreated), and basophilia was present in 6 (50%). Immunophenotypic studies showed that the blasts were positive for CD9, CD13, CD33, CD38, CD117, and HLA-DR in all cases assessed. CD34 was positive in 11 (92%) of 12, and terminal deoxynucleotidyl transferase was positive in 7 (64%) of 11 cases. The t(6;9) was the only cytogenetic abnormality detected in 7 cases (58%), and 5 cases had additional chromosomal abnormalities. Of 8 cases assessed, 7 (88%) had flt3 gene mutations. We conclude that t(6;9) positive AML cases have distinctive morphologic features, an immunophenotype suggesting origin from an early hematopoietic progenitor cell, and a high frequency of flt3 gene mutations. PMID- 15362365 TI - Gaucher cells demonstrate a distinct macrophage phenotype and resemble alternatively activated macrophages. AB - Although the existence of anti-inflammatory alternatively activated macrophages (aamphi) has been accepted widely based on in vitro studies, their in vivo location, phenotype, and function still are debated. Gaucher disease (GD) is caused by a genetic deficiency in the lysosomal enzyme glucocerebrosidase and is characterized by accumulation of glycosphingolipids in so-called Gaucher cells (GCs). By using immunohistochemical analysis, we investigated whether this results in an aamphi phenotype. GCs are macrophage-like cells, expressing acid phosphatase, CD68, CD14, and HLA class II, but not CD11b, CD40, or dendritic cell markers. GCs show infrequent immunoreactivity for mannose receptor GCs did not express proinflammatory cytokines such as tumor necrosis factor alpha and monocyte chemoattractant protein 1, but did express the aamphi markers CD163, CCL18, and interleukin-1 receptor antagonist. Furthermore, CD36 and signal receptor protein alpha, involved in lipid uptake, also were observed on GCs. Thus, GCs represent a distinctive population of myeloid cells that resemble aamphi but differ from previously described in vitro aamphi. PMID- 15362366 TI - Gleason grading of prostate cancer: level of concordance between pathologists at the University Hospital of the West Indies. AB - Our aim was to study the level of interobserver concordance in the Gleason scores of prostate needle biopsy specimens reported at 1 institution. A retrospective review of all prostate needle biopsy specimens in which a diagnosis of adenocarcinoma was made during the year 2000 was conducted. Parameters evaluated included the Gleason score, Gleason grades identified, the percentage of Gleason grades 4 and 5, and the percentage of tumor in the biopsy specimen. Our results demonstrated a 60% overall concordance in consensus Gleason scores, which increased to 80% when considered in groups of a Gleason score of less than 7 vs 7 or more. The greatest discordance seemed to be in distinguishing Gleason score 6 from 7 and was more frequent among biopsy specimens with lower tumor volumes, particularly among those with less than 30% involvement. A small percentage of Gleason grade 4 pattern might predict disagreement as well. Strategies for improving accuracy of Gleason score 7 should be devised, and consensus diagnosis for biopsy specimens that demonstrate a low percentage of tumor volume is recommended. PMID- 15362367 TI - Concurrent evaluation of p53, beta-catenin, and alpha-fetoprotein expression in human hepatocellular carcinoma. AB - Recent models suggest that hepatocellular carcinoma (HCC) develops through several independent pathways marked by key mutations in the beta-catenin or p53 gene. An additional pathway potentially is marked by aberrant expression of a fetoprotein (AFP). To see whether these potential markers are expressed independently, we immunostained sequential sections from 55 HCCs. Of the cases, 30 (55%) were positive for 1 or more proteins: AFP, 19 cases (35%); p53, 12 cases (22%); and beta-catenin, 9 cases (16%). Seven tumors (13%) were positive for more than 1 protein, with 4 of 7 positive in the same area of tumor and 3 of 7 positive in different areas of the carcinomas. By statistical analysis, expression of the markers was independent of one another and of tumor size. Concurrent evaluation of p53, beta-catenin, and AFP protein expression showed no associations, supporting models in which these proteins might serve as markers of independent pathways in the development of HCC. PMID- 15362368 TI - Efficacy of cone biopsy of the uterine cervix during frozen section for the evaluation of cervical intraepithelial neoplasia grade 3. AB - We retrospectively selected 22 cases in which patients with a biopsy-proven diagnosis of cervical intraepithelial neoplasia grade 3 underwent cervical conization for frozen section (FS) evaluation followed by hysterectomy at the University of California Irvine Medical Center, Orange, during the August 1995 to September 9, 2001. All slides from FS and permanent section (PS) and hysterectomy specimens were reviewed. FS diagnoses were compared with those of previous biopsies, PS, and hysterectomy specimens. The PS correlated with FS in all cases but 1. Appropriate surgery was performed for all patients based on FS diagnosis. The McNemar test was used to compare the results of FS and PS, with a 2-sided P value of 1.0 and a c coefficient of 0.7755 with a 95% confidence level, indicating that the 2 groups were not significantly different. FS evaluation of cervical conization is as efficacious and accurate as evaluation of regular specimens in providing information for the appropriateness of same-day surgery. We recommend that entire tissue be submitted for FS to avoid sampling errors and to increase diagnostic accuracy. PMID- 15362369 TI - Defective mismatch-repair colorectal cancer: clinicopathologic characteristics and usefulness of immunohistochemical analysis for diagnosis. AB - The purpose of our study was to determine the usefulness of immunohistochemical analysis for the diagnosis of mismatch-repair (MMR) gene defective colorectal tumors and to describe their prevalence and clinicopathologic characteristics. We studied 172 cases. DNA was extracted from formalin-fixed, paraffin-embedded surgical samples, and microsatellite analysis was performed by polymerase chain reaction with BAT-26. The results were correlated with immunohistochemical analysis for hMLH1 and hMSH2. Microsatellite instability (MSI) was detected in 13 (7.6%) tumors, and all showed loss of protein expression of hMLH1 (11/13) or hMSH2 (2/13) (P < .000). Patients with MMR-defective tumors more frequently had poorly differentiated tumors (5/13 [38%] vs 18/159 [11.3%]; P = .02) located in the ascending colon (8/13 [62%] vs 30/159 [18.9%]; P < .0001) and a personal history of other neoplasms (4/13 [31%] vs 18/159 [11.3%]; P = .05). There were no differences in age, family history of cancer, or TNM stage. Immunohistochemical analysis seems to be a reliable method to detect most colorectal cancers with defective MMR genes. PMID- 15362370 TI - Differential expression of superoxide dismutases in lung cancer. AB - Oxidant-antioxidant balance is known to regulate growth factors and invasion of tumor cells. Manganese superoxide dismutase (MnSOD), copper zinc SOD (CuZnSOD), and extracellular SOD (ECSOD), the first-line antioxidant defenses, were studied in lung carcinomas by immunohistochemical analysis (n = 139, 56, and 37, respectively) and in 8 lung tumor specimens by Western blot analysis and SOD activity measurement. Altogether, 49% of squamous cell carcinomas and 43% of the adenocarcinomas were positive for MnSOD by immunohistochemical analysis; corresponding values for CuZnSOD were 79% and 93%, respectively. MnSOD and CuZnSOD by Western blot analysis were 27% and 22% higher, and CuZnSOD activity was 93% higher (P = .06) in carcinomas than in nonmalignant lung tissue samples. ECSOD, a mainly extracellular enzyme, showed weak positivity only in 4 of 37 carcinomas, and by Western blot analysis showed 70% lower immunoreactivity (P < .0001) than in nonmalignant lung tissue samples. It is highly likely that low expression of ECSOD might have fundamental effects on the extracellular redox state of lung tumors with potential consequences on tumor behavior. PMID- 15362371 TI - Paucicellular and asymptomatic lymphocytic colitis: expanding the clinicopathologic spectrum of lymphocytic colitis. AB - We examined clinicopathologic associations and biopsy changes that suggested classic lymphocytic colitis (C-LC) but were less well developed in intensity or distribution in 19 cases, which we termed paucicellular LC (P-LC). We also studied clinicopathologic associations and prevalence of LC in 100 asymptomatic, non-gluten-sensitive adults who underwent screening surveillance colonoscopy for previous adenoma. The control group was 38 randomly selected morphologically C-LC cases. The features of P-LC were foci of mildly increased lamina propria lymphoplasmacytic inflammation and increased surface intraepithelial lymphocytes separated by foci or tissue fragments of normal mucosa. Mean age and rates of female sex, endoscopically normal appearing colon, abdominal pain, watery stools, weight loss, connective tissue diseases, and consistent ingestion of nonsteroidal anti-inflammatory drugs (NSAIDs) were similar for P-LC and C-LC patients. Of 100 asymptomatic patients, 26 (26%) had LC and 43 (43%) used NSAIDs daily. Of these 43 patients, 14 (33%) had P-LC or C-LC. Daily NSAID ingestion was associated significantly with LC (P = .024). P-LC patients had clinicopathologic relationships similar to those of C-LC patients, suggesting they should be considered part of the morphologic spectrum of LC. LC in asymptomatic adults might be more common than previously thought and might not be associated with watery diarrhea syndrome. PMID- 15362372 TI - Expression of activated TrkA protein in melanocytic tumors: relationship to cell proliferation and clinical outcome. AB - We evaluated expression of activated nerve growth factor receptor tyrosine kinase (p-TrkA) by immunohistochemical analysis in 152 primary and 64 metastatic human melanoma biopsy specimens and 8 nevi. Membranous, cytoplasmic, and/or nuclear expression of p-TrkA was seen in 54.6% of primary melanomas and 30% of metastases. Membranous p-TrkA was detected in 21.7% of primary and 14% of metastatic melanomas and cytoplasmic immunoreactivity in 28.9% of primary tumors and in 22% of metastases. Significantly fewer metastases than primary tumors expressed nuclear p-TrkA (16% vs 39.5%; P = .006). A significantly higher percentage of nodular than superficial spreading melanomas expressed membranous (40% vs 11%; P < .0001) p-TrkA. Nevi expressed no membranous or cytoplasmic p TrkA; 63% showed nuclear reactivity. p-TrkA expression varied significantly with thickness of primary tumors (lower expression in thinner lesions: membranous, P = .004; cytoplasmic, P = .001; nuclear, P = .031). An association between ulceration and membranous (P = .054), cytoplasmic (P < .0001), and nuclear (P = .022) p-TrkA expression was found. Membranous p-TrkA significantly predicted decreased overall survival (P = .002). A significant association between membranous p-TrkA and cyclin A (P = .004) and Ki-67 (P < .0001) and between cytoplasmic p-TrkA and cyclin A (P < .0001), Ki-67 (P = .004), and cyclin D3 (P = .027) was found. p-TrkA had no effect on MAPK(ERK1/2) activation. A significant inverse association between cytoplasmic beta-catenin and cytoplasmic p-TrkA levels (P = .006) and between nuclear p-TrkA and cytoplasmic E-cadherin (P = .022) was seen. We present the first evidence of a role for TrkA activation in a subset of melanomas as a predictor of an aggressive phenotype and poor outcome. PMID- 15362373 TI - Usefulness of Cdx2 in separating mucinous bronchioloalveolar adenocarcinoma of the lung from metastatic mucinous colorectal adenocarcinoma. AB - We studied the diagnostic value of Cdx2 to distinguish mucinous bronchioloalveolar carcinoma from mucinous colorectal adenocarcinoma metastatic to the lung. We retrieved 92 via the hospital computer system, including 30 mucinous bronchioloalveolar carcinomas, 32 nonmucinous bronchioloalveolar carcinomas, and 30 mucinous colorectal adenocarcinomas metastatic to the lung. All cases were confirmed by clinical history and surgical resection with occasional immunohistochemical studies. Cases were stained with antibodies against Cdx2, thyroid transcription factor-1 (TTF-1), cytokeratin (CK) 7, and CK20. Bronchioloalveolar carcinoma, mucinous type, showed positive staining for Cdx2, TTF-1, CK7, and CK20 in 0 (0%), 5 (17%), 30 (100%), and 18 (60%) of 30 cases, respectively; nonmucinous tumors were positive in 0 (0%), 30 (94%), 32 (100%), and 0 (0%) of 32 cases, respectively. For colorectal adenocarcinoma, the positive staining for Cdx-2, TTF-1, CK7, and CK20 was 29 (97%), 0 (0%), 7 (23%), and 29 (97%) of 30 cases, respectively. Our results demonstrated Cdx2 as a sensitive and specific marker for differentiating metastatic colorectal adenocarcinoma from mucinous bronchioloalveolar adenocarcinoma. PMID- 15362374 TI - Carriage of the apolipoprotein E-epsilon4 allele and histologic outcome of recurrent hepatitis C after antiviral treatment. AB - Carriage of the epsilon4 allelic variant of the apolipoprotein E (ApoE) gene might affect the outcome of hepatitis C virus (HCV) infection. The liver transplantation setting offers the opportunity to verify the role of the donor's vs recipient's ApoE polymorphism. Twenty-four patients (16 men) with recurrent hepatitis C, all infected by HCV-1b and treated with interferon and ribavirin, were genotyped for ApoE variants. Liver biopsies were done at baseline and 12 months later After treatment, staging scores improved in 10 of 24 patients. Staging improvement was associated with recipient sex, completion of the full antiviral schedule, and recipient's epsilon4 carriage. The beneficial effect of epsilon4 carriage toward the progression of fibrosis was due entirely to the contribution given by male patients and was independent of the viral response. Recipients', but not donors', carriage of at least 1 epsilon4 allele might be associated with a better histologic outcome in recurrent HCV infection. PMID- 15362375 TI - An abnormal cervicovaginal cytology smear in uterine carcinosarcoma is an adverse prognostic sign: analysis of 25 cases. AB - Carcinosarcoma of the uterus has been poorly characterized on cervicovaginal (Pap) smears, and we examine whether they effectively screen for carcinosarcoma and whether an abnormal Pap smear result has any clinical importance. Twenty-five patients with histologically confirmed carcinosarcoma had a conventional Pap smear shortly before diagnosis. Eleven smears (44%) originally were read as abnormal (malignant or atypical), and 4 additional cases were read as abnormal on retrospective review (15/25 [60%]). All malignant elements were epithelial, and 2 cases (8%) had atypical spindle cells, but no diagnostic sarcoma. Cervical involvement was the only histologic parameter correlating with an abnormal Pap smear result (P = .04). Univariate analysis found stage III or IV disease was an adverse prognostic sign compared with stage I or II disease (mean survival, 8 vs 36 months, respectively; P = .001), and multivariate analysis indicated that an abnormal Pap smear result correlated with worse survival (P = .023). The conventional Pap smear is insensitive (60%) for detecting carcinosarcoma, but when the result is abnormal, the Pap is an important stage-independent adverse prognosticator. PMID- 15362376 TI - Papillary lesions of the breast with and without atypical ductal hyperplasia: can we accurately predict benign behavior from core needle biopsy? AB - Evaluation of papillary lesions of the breast can be difficult, and in core needle biopsy specimens, accurate diagnosis is challenging. Initial studies suggested that all papillary lesions revealed by core biopsy required surgical excision. Recent data suggest that only papillary lesions with atypical ductal hyperplasia (ADH) revealed by core biopsy need surgical excision. We evaluated our experience at the University of Washington Medical Center, Seattle, with papillary lesions with and without ADH on core biopsy to determine whether diagnostic accuracy can be achieved. In 51 core biopsy specimens, we evaluated the presence or absence of ADH: 25 were benign papillomas; 26 were papillomas with ADH. Surgical follow-up was available for 36 cases (11 papillomas and 25 papillomas with ADH). Clinical (radiologic) follow-up was available in 5 papilloma cases (average follow-up, 35.6 months). Follow-up revealed that all papillomas on core biopsy were benign. Excisional biopsy revealed ductal carcinoma in situ or invasive carcinoma in 12 (48%) of 25 papillary lesions with ADH. Benign papillomas can be adequately diagnosed with core biopsy. All papillary lesions with ADH require surgical excision owing to the high rate of associated neoplasia. PMID- 15362377 TI - Prognostic factors in survival of patients with stage Ta and T1 bladder urothelial tumors: the role of G1-S modulators (p53, p21Waf1, p27Kip1, cyclin D1, and cyclin D3), proliferation index, and clinicopathologic parameters. AB - We studied 159 cases of superficial (stage Ta or T1) bladder tumors to determine the significance on survival of a subset of regulators of transition from G1 to S phase of the cell cycle (p53, p21Waf1, p27Kip1, cyclin D1, cyclin D3) and tumor proliferation (Ki-67 [MIB-1]). Clinical findings (patient age, sex, tumor size, grade, stage [Ta or T1]) were included in the analysis. Univariate analysis revealed association of tumor size (P = .0353), grade in stage Ta tumors (P = .0074), cyclin D1 expression (P = .0182), and Ki-67 index (P = .0033) with disease-free survival and of tumor size (P = .0005), stage (P = .0494), cyclin D3 expression (P = .0105), and Ki-67 index (P = .0272) with overall survival. Cox multivariate analysis revealed cyclin D1 expression and high proliferation index (disease-free) and tumor size, cyclin D3 expression, and high proliferation index (overall survival) as independent predictors. Results suggest that alterations of the progression from the G1 to S phase of the cell cycle are common in papillary urothelial bladder tumors. High tumor proliferation, expression of cyclins D1 and D3, and tumor size at diagnosis might be relevant predictors of survival in patients with stage Ta and T1 bladder urothelial tumors. PMID- 15362378 TI - SARS viral RNA. PMID- 15362379 TI - Development of an ethanol model using social insects: IV. Influence of ethanol on the aggression of Africanized honey bees (Apis mellifera L.). AB - Experiments were designed to determine whether ethanol influenced aggression in honey bees. Two experiments are reported. In Exp. 1, harnessed honey bees were fed a 1%, 5%, 10%, or 20% ethanol solution. Two control groups received either a sucrose solution only or no pretreatment, respectively. The dependent variable was the number of sting extensions over 10 min. Analysis showed that aggression in harnessed bees was not influenced by prior ethanol consumption. Because there was some suspicion that the extension of the sting apparatus may be hindered by harnessing, and the authors wanted to use a design that increased ecological validity, Exp. 2 was conducted with free-flying bees. Sucrose or 20% ethanol solutions were placed in front of beehives, and the number of stings on a leather patch dangled in front of the hive served as the dependent variable. The experiment was terminated after 5 hr. because bees exposed to ethanol became dangerously aggressive. A unique aspect of the study was that Africanized honey bees were used. PMID- 15362380 TI - Conflicts, workplace bullying and burnout problems among municipal employees. AB - The psychosocial workplace environment among the municipal employees of a Finnish town with 57,000 inhabitants was investigated. The prevalence of both self experienced and observed cases of conflicts, burnout, and workplace harassment (bullying) was measured. Altogether, 1,961 employees (1,491 women and 309 men) whose mean age was 42.6 yr. (SD = 9.8) participated in the study. Sex, age, and workplace differences were found. Of the respondents, 27% had experienced difficult or extreme conflicts, 29% experienced burnout, 50% had seen at least one colleague experience burnout, 16% felt they had been bullied, and 37% had seen a colleague being bullied at work. Men reported experiencing more conflicts than women. The frequency of reporting burnout symptoms and bullying was associated with age. Strong correlations were found among the various problem areas. PMID- 15362381 TI - Admission to university engineering programs in Sweden: a multipurpose approach. AB - This brief article provides a description of some new ideas about admission of university engineering students in Sweden. The current system of admission is based on upper-secondary school grades and the Swedish Scholastic Assessment Test. These measures are used for admission to all higher education. For many reasons, ideas for a new admission model have been proposed. This model includes a sector-oriented admission test, which the universities are supposed to use for different purposes, such as selection, eligibility, diagnostics, and recruitment. PMID- 15362382 TI - Role of suggestion in odor-induced mood change. AB - The effects of ambient odor (lavender, neroli or placebo) and suggestions related to the effects of an odor (relaxing, stimulating or none) on mood were explored. Mood of 90 undergraduate women was assessed using physiological measures (heart rate and skin conductance) and the self-report Profile of Mood States questionnaire. Analysis indicated that physiological measures were influenced by suggestion in predictable directions. Relaxing odors yielded decreases in heart rate and skin conductance, with stimulating odors yielding the reverse effects under equivalent conditions. These data further support the notion that expectations play a significant role in mediating odor-evoked mood changes. PMID- 15362383 TI - A low-cost drinkometer circuit suitable for insects and other organisms. AB - A low-cost drinkometer circuit suitable for use with insects is described. The circuit is composed of readily available components powered by a battery pack. Schematic diagrams of the circuit and simulations are provided. The operational theory behind the development of the circuit is discussed. The usefulness of the circuit is illustrated by a simple consumption experiment in which the amount of sucrose consumed by honey bees is determined. PMID- 15362384 TI - Adolescents' peer-rated mental health, peer-acceptance, and irrational beliefs. AB - Based on the notion of positive psychology, the interrelationships among peer rated mental health, social acceptance and irrational beliefs were examined using Korean youth. We predicted that adolescents who were rated by their peers to be relatively more mentally healthy, compared to those who were rated to be less mentally healthy, would (a) receive higher social acceptance from peers and (b) report fewer irrational beliefs. 476 adolescents (252 boys and 224 girls; M age = 16.7 yr., SD=.46), all recruited from a single high school in an upper middle class area in Seoul, Korea, rated each others' mental health in five areas of mental health, i.e., concern for others, contentment with life, creativity and industry, good interpersonal relationships and ability to cope with stressful situations and social acceptance and responded to a questionnaire measuring irrational beliefs. Analysis showed that those adolescents who were rated by their peers to be more mentally healthy received higher social acceptance from those peers than those rated as less mentally healthy; however, this effect size was small. Implications of the study are discussed. PMID- 15362385 TI - Imagery factors in the Spanish version of the Verbalizer-Visualizer Questionnaire. AB - The factor structure and internal consistency reliability of the Spanish version of the Verbalizer-Visualizer Questionnaire was analyzed. 969 male and female high school students completed the questionnaire and the Gordon Test of Visual Imagery Control. Factor analysis indicated that the questionnaire comprises five factors: "interest in words," "dream vividness and frequency," "verbal fluency," "task performance difficulty," and "ways of thinking and acting." The internal consistency was very low (Cronbach alpha = .30). The correlation between scores and those on the Gordon test was statistically significant because N was large but very weak (r =.08, p<.05). The questionnaire measured five distinct factors, and this should be taken into account when using the Spanish version rather than a coherent dimension. PMID- 15362386 TI - Associations among homesickness, anger, anxiety, and depression. AB - The study investigated the association of homesickness with the related concepts of anxiety, depression, and anger. Two conceptualizations of homesickness were used, one considering homesickness as a state, characterized by severe symptoms of a depressive episode, and the other one as a self-reported tendency to experience homesickness in times of separation from the familiar environment. The latter conceptualization can rather be considered as an enduring trait and does not specifically refer to actual feelings of depression. We compared these two conceptualizations of homesickness in a random sample from the Dutch population (n=485) by assessing their uni- and multivariate associations with anger, anxiety, depression, and anxiety-sensitivity. Both conceptualizations of homesickness were associated with anxiety and depression and with the externalization of anger. No association was found between internalization of anger or control over internalized and externalized anger for either conceptualization of homesickness. The pattern of multivariate associations between homesickness and its emotional correlates was similar for both conceptualizations, although homesickness as a state appeared to have a stronger association with depression, whereas the tendency to develop homesickness showed a particularly strong association with anxiety. It is concluded that homesickness can be considered as a mixed emotion of anxiety and depression ("Cothymia"), but that depression is more characteristic of homesickness as a state, whereas anxiety is more important if homesickness is conceptualized as an enduring tendency. PMID- 15362387 TI - Convergent validity of the Templer, Collett-Lester, and Arabic Death Anxiety Scales: rejoinder. AB - The Templer Death Anxiety Scale, the Arabic Scale of Death Anxiety by Abdel Khalek, and the Collett-Lester Fear of Death Scale were administered to a convenient sample of 81 male and female Kuwaiti undergraduates enrolled in social science courses (M age=22.0 yr., SD=2.3). Pearson correlations between the total scores were significant and positive. Only one high-loaded factor was extracted and labeled General Death Anxiety, indicating good convergent and factorial validity of these scales. PMID- 15362388 TI - An exploratory study of differences in views of factors affecting sexual orientation for a sample of lesbians and gay men. AB - An exploratory study of lesbians (70) and gay men (118) from a rural state in the mid-South was conducted using a self-administered, mail-out survey. The nonrandom sample was drawn from organizational mailing lists, snowball sampling, and a convenience sample at a community event. Respondents were asked to indicate the extent to which each of the following affected sexual orientation: genetics, relationship between parents, relationship with parents, birth order, peers, growing up in a dysfunctional family, growing up in a single-parent family, negative experiences with the opposite sex, and positive experiences with the same sex. Similar to studies of heterosexual men and women, these gay men were more likely to view sexual orientation as a result of genetics than the lesbian respondents. Further, the lesbian group were more likely to view positive relationships with the same sex to have a great influence on sexual orientation. These data indicate there are sex differences in views on factors that affect sexual orientation. PMID- 15362389 TI - Cognitive and personality factors associated with assaultive and domestic offenders. AB - An evaluation of domestic assaultive (n=46), general assaultive (n= 23) and nonassaultive (19 property offenders) groups was performed to compare cognitive (Test of Nonverbal Intelligence), personality (Minnesota Multiphasic Personality Inventory), aggression (Buss-Durkee Hostility Inventory), and criminal sentiments (Carlson Psychological Survey). Analysis showed minimal significant differences amongst the groups. A significant difference was found for the Verbal Hostility subscale of the Buss-Durkee Hostility Inventory. The domestic assaultive offenders were similar to the general assaultive and nonviolent offenders, with a greater disparity for the control group. Furthermore, a significant difference was noted for the Hypochondriasis scale of the Minnesota Multiphasic Personality Inventory. The general assaultive and control (n = 15) groups had similar scores, with the greatest difference found for nonviolent offenders who were least anxious. Discriminant analysis showed group membership could be identified for 89.1% of the domestic assaultive group. PMID- 15362390 TI - Comments on Dr. Walter R Schumm's paper "What was really learned from Tasker and Golombok's (1995) study of lesbian and single parent mothers?". AB - Comments on Schumm's analysis and evaluation of the 1995 report by Tasker and Golombok focus on insights gained during research and requirements for drawing conclusions: larger samples, more sophisticated methods, appropriate comparative cohorts, and longitudinal studies. PMID- 15362391 TI - Ideal gender identity related to parent images and locus of control: Jungian and social learning perspectives. AB - In this research, we wanted to clarify how gender images are different or invariant and related to parents, attributes, and the attitude of controlling life (locus of control) in two cultural contexts, Japan and Finland. For this purpose, students' ideal gender images, consisting of ideal mother, female, father and male images, and parents' similarity to the four ideal gender images were studied in 135 Japanese and 119 Finnish university students. Major findings were (a) Japanese students' ideal gender images were more stereotypic than those of Finnish students; (b) students' ideal mother image and parents' similarity to the ideal mother image were related only to their sex, which supports Jung's theory; (c) students socially learned other ideal gender images, but these did not fit with expectation from social learning theory; (d) Japanese students' mothers are models or examples of gender images, but Finnish male students did not seem to base their ideal gender images on their parents. Implication of measures was discussed. PMID- 15362392 TI - Relations of motivational beliefs and self-regulated learning outcomes for Iranian college students. AB - The present study examined the correlations of scores for motivational beliefs and self-regulated learning outcomes among 200 college students who responded to Motivational Strategies for Learning Questionnaire. Analysis showed a positive correlation between scores for motivational beliefs and self-regulated learning outcomes but a negative one for scores on test anxiety and self-regulated learning. PMID- 15362393 TI - Does sex of client affect counselors' evaluation? AB - This study examined whether clients' positive or negative self-disclosure and sex affected counselors' evaluation of the clients. 80 counselors (30 certified, 45 master's and five doctoral trainees; six men and 74 women), after viewing a video taped counseling interview in which the client self-disclosed either largely positive or negative contents responded to the Clinical Impression Scale which contained 8 clinical characteristics of clients (assertiveness, overall coping skills, anxiety, attitude toward counseling, motivation to change, self-esteem, insight, and depression). When the client self-disclosed largely negative content about self, counselors evaluated male client more negatively, i.e., less assertive, poor coping skills, higher anxiety, more negative attitude toward counseling, less eager to change, lower self-esteem, less insight, and higher depression) than female client. However, when the client self-disclosed largely positive content, there was no statistically significant sex difference between the two groups. The implications are discussed. PMID- 15362394 TI - Death anxiety, death depression, and death obsession: a general factor of death distress is evident: a reply. AB - A sample of 75 (16 men, and 59 women) Kuwaiti college students responded to Templer's and Collett-Lester Death Anxiety Scales, Templer, et al.'s Death Depression Scale and Abdel-Khalek's Death Obsession Scale. A general high-loaded factor of death distress was extracted using the total scores. However, in using the Collett-Lester four subscales, the Fear of Death and Dying of Others loaded on a second factor. PMID- 15362395 TI - Effects of inferred social status and a beginning driver's sticker upon aggression of drivers in Japan. AB - The present study examined how inferred social status and a beginning driver's sticker influenced aggressive drivers' behavior on the road in Japan. A 2 x 2 x 2 factorial design was constructed to examine the effects of social status of an experimental car (high or low), with a beginning driver's sticker or no beginning driver's sticker, and a male or female driver. Analysis showed that horn-honking latency in the sticker condition was longer than that in the no-sticker condition in the low status car. In the high status car, the horn was sounded sooner in the condition with a beginning driver's sticker. A beginning driver's sticker promoted aggressive behavior oward a high status person, whereas it inhibited aggressive behavior toward low status individuals. Thus, response to a beginning driver's sticker appears to vary according to inferred social status. PMID- 15362396 TI - Classical conditioning of proboscis extension in harnessed Africanized honey bee queens (Apis mellifera L.). AB - Experiments are reported on learning in virgin Africanized honey bee queens (Apis mellifera L.). Queens restrained in a "Pavlovian harness" received a pairing of hexanal odor with a 1.8-M feeding of sucrose solution. Compared to explicitly unpaired controls, acquisition was rapid in reaching about 90%. Acquisition was also rapid in queens receiving an unconditioned stimulus of "bee candy" or an unconditioned stimulus administered by worker bees. During extinction the conditioned response declines. The steepest decline was observed in queens receiving an unconditioned stimulus of bee candy. These findings extend previous work on learning of Afrianized honey bee workers to a population of queen bees. PMID- 15362397 TI - Does social desirability inflate the correlation between self-esteem and anxiety? AB - Although several authors concluded that self-esteem is negatively related to anxiety, it is an open question what role social desirability plays in this relation. In a sample of 61 German students, the correlation between self-esteem and anxiety was significantly reduced from -.59 to -.51 when social desirability was partialed out. Thus, social desirability does seem to inflate the correlation between self-esteem and anxiety when both constructs are measured with self reports. Yet, this effect of social desirability appears so small that it is probably negligible for most research purposes. PMID- 15362398 TI - Whoever is influential is creative: how Chinese undergraduates choose creative people in Chinese societies. AB - This study surveyed 994 undergraduates in Guangzhou, Hong Kong, Nanchang, Nanjing and Xian about their nomination and evaluation of the most creative people in Chinese societies as well as their valuation of collectivistic and individualistic goals of creativity. Politicians and scientists/inventors were mostly nominated and were generally rated higher on social contribution than on creativity. Artists/musicians and writers/poets were rarely nominated and were mostly rated higher on creativity than on social contribution. Collectivistic goals of creativity were significantly more valued than individualistic goals of creativity. There appears to be a social validation of creativity such that meritorious salience of creativity and social influence of the creator were greatly emphasized. Merit-based attribution of creators may have a detrimental effect on perception and promotion of general creativity in Chinese societies. PMID- 15362399 TI - Daily class objectives and instructor's effectiveness as perceived by students. AB - Course and instructor evaluations completed by students (N=255) enrolled in multiple discussion sections of one introductory-level college class, led by the same instructor over four consecutive semesters, indicated a positive correlation (r = .34, p<.001) between awareness of daily class objectives and ratings of the instructor's overall teaching effectiveness. Students for whom the presentation of daily objectives was memorable or important were more likely to perceive the instructor's teaching as effective, at least as measured by students' standardized evaluations. PMID- 15362400 TI - The primacy of beauty in judging the aesthetics of objects. AB - The conceptual structure of the aesthetics of objects was investigated. To this end, associative namings for the word "aesthetics" were collected from 311 nonartist German college students in a timed verbal association task. 590 different adjectives were produced, depicting diversification of the concept. The adjective "beautiful" was given by more than 90% of the participants. The adjective "ugly" was the second most frequent naming, used by almost half of the students. All other namings were markedly less frequently produced. It is argues that the beautiful-ugly dimension represents the primary concept in the aesthetics of objects, so that performing aesthetic judgments of the beauty of objects comes naturally to individuals. In other words, the most prototypical aesthetic judgments are those of beauty. Furthermore, the majority of generated words had a positive valence as measured by an additional valence-rating study including 41 participants. This result contrasts with comparable studies of emotion terms, as such studies typically show a negativity bias. Frequency in general language use and valence of the adjectives did not account for the results. PMID- 15362401 TI - Relationships between locus of control and paranormal beliefs. AB - The present study investigated the associations between scores on paranormal beliefs, locus of control, and certain psychological processes such as affect and cognitions as measured by the Linguistic Inquiry and Word Count. Analysis yielded significant correlations between scores on Locus of Control and two subscales of Tobacyk's (1988) Revised Paranormal Beliefs Scale, New Age Philosophy and Traditional Paranormal Beliefs. A step-wise multiple regression analysis indicated that Locus of Control was significantly related to New Age Philosophy. Other correlations were found between Tobacyk's subscales, Locus of Control, and three processes measured by the Linguistic Inquiry and Word Count. PMID- 15362402 TI - Credit card use and prefrontal cortex dysfunction: a study in neuroeconomics. AB - In a study of 100 students, no associations were found between a measure of the prefrontal system dysfunction personality with credit card use and attitudes. PMID- 15362403 TI - Children's somatization symptoms: correlations with trait anxiety, anxiety sensitivity, and learning experiences. AB - The objectives of this study were to examine associations between trait anxiety, anxiety sensitivity, and learning experiences, on the one hand, and children's somatization symptoms, on the other hand. A sample of 190 nonclinical children completed the trait anxiety scale of the State-Trait Anxiety Inventory for Children, the Childhood Anxiety Sensitivity Index, and the Children's Somatization Inventory, and were interviewed about learning experiences in relation to physical symptoms. Correlational and regression analyses indicated that trait anxiety, anxiety sensitivity, and parents' anxiety-encouraging behaviours in relation to somatic symptoms were significantly positively associated with somatization. These results indicate that there might be common vulnerability factors in childhood anxiety and somatization symptoms. PMID- 15362404 TI - Postpartum depression following normal vaginal delivery among Nigerian women. AB - The study investigated postpartum depression among Nigerian women by comparing 83 mothers who had normal vaginal deliveries with 83 matched controls. Analysis of scores on the Zung Self-rating Depression Scale indicated that the mothers and the controls were not significantly different on depression during the immediate postpartum period. There were significant relationships between postpartum depression and education after birth, not having male children at after 6 wk., primiparity after birth, and puerperal complications. PMID- 15362405 TI - Effect of type of survey administrator on Consumer Assessment of Care. AB - This study evaluated the possible effects of peer, unit staff, and quality management staff survey administrators on item means of the inpatient version of the Consumer Assessment of Care, a 14-item mental health service satisfaction survey developed by the New York State Office of Mental Health and administered at South Beach Psychiatric Center. Of 511 inpatients who completed the survey, 215 were administered by unit staff, 156 by consumers, and 140 by quality management staff. Each subgroup used a mixed-mode administration method for consecutive 6-mo. periods. Reliability and factor analysis indicated the survey was psychometrically robust. Item means for the types of administrator were significantly different. Tukey HSD multiple-comparisons indicated that item means were significantly higher for unit staff administrators than peer and quality management staff administrators, which did not differ from each other. Response set, acquiescence, or socially desirable responding may elicit higher item means for unit staff administrators. PMID- 15362406 TI - Sex differences in persuadability of human and computer-synthesized speech: meta analysis of seven studies. AB - To examine sex differences in persuasiveness, we conducted a meta-analysis of seven studies from our laboratory on reactions to human versus computer synthesized speech. We tested three hypotheses: (1) people would be more persuaded by human speech than by computer-synthesized speech, (2) women would be slightly more persuaded than men, and (3) the sex difference would be more pronounced for human speech than for synthetic speech. While there was support for the first two hypotheses, there was none for the third. Also, no consistent support was found for a moderating effect of mode of presentation, audio versus video. PMID- 15362407 TI - Dating and commitment choices as a function of ethnicity among American college students in California. AB - The incidence of interracial and interethnic dating and marriage in the United States has increased. This investigation examined dating and commitment choices as a function of ethnicity and sex among groups of Euro-American, Hispanic American, Asian-American, and African-American college students. A convenience sample of college students comprising 329 heterosexual subjects (134 men, 195 women) was surveyed regarding their partner preferences for dating, visiting parents, marriage, and bearing children. It was hypothesized that subjects would consider dating partners from different ethnic groups, but when making a commitment to marriage and children would prefer members of their own group. This hypothesis was supported in half of the groups: Euro-American men, African American men, Asian-American women, and African-American women. A discussion of dating and commitment choices among ethnic and sex groups is presented and discussed. PMID- 15362408 TI - Quality control pharmacy tasks: big five personality model and accuracy of error detection. AB - The relationships between extraversion, conscientiousness, neuroticism, and performance were examined in the context of an error detection task completed under stable and shifting workload conditions. 114 participants checked two sets of 40 fictitious prescriptions for errors. Errors were inserted at rates varying from 26% to 38%. Workload was manipulated by altering the amount of time allowed to complete each set. The stable workload group had 45 min. per set, the workload up-shift group had 60 min. for the first set and 30 min. for the second set. Performance was measured using hit rates and false alarms. Analysis indicated that extraversion and conscientiousness were correlated with hit rates, but only in the stable workload condition. Results are discussed and research directions are considered. PMID- 15362409 TI - The PC anxiety scale: a cross-cultural replication. AB - A brief 6-item scale to measure anxiety about computers, developed using American students, showed good internal consistency, reliability and construct validity for British students (77 men, 65 women). PMID- 15362410 TI - Psychopathology and mental retardation: a study using the Rorschach Inkblot Test. AB - The aim of this research was to study the psychological effects of disorders such as schizophrenia and depression associated with mental retardation. The Rorschach Inkblot Test and the Wechsler Adult Intelligence Scale were administered to a group of 97 subjects (52 women and 45 men) ages 15:10 yr. to 36:6 yr. (M=21:5, SD=5:3). The subjects were divided into four subgroups according to the presence or absence of mental retardation and psychiatric diagnosis (schizophrenia versus depression). The quality of the perception in Rorschach responses and the ErlebnisTypus scores differentiated psychotic and depressed subjects well. These disorders, when associated with mental retardation, make impairment of perceptual performance worse. The interaction between Axis I mental disorders (according to DSM-IV diagnosis) and mental retardation, an Axis II disorder, is discussed. PMID- 15362411 TI - Associations of sex and type of relationship on intimacy. AB - Sex differences in self-perceived intimacy were investigated in both same-sex friendships and romantic relationships. Men and women had significantly different means on intimacy with their same-sex friends but reported similar means with romantic partners. Both men and women reported significantly higher mean scores on intimacy with romantic partners than with friends. PMID- 15362412 TI - Nonconscious idea generation. AB - The recognition of the correct solution to a problem after a period when one is not actively searching for an answer is well documented. However, previous research has focused on problems an individual has not yet resolved. We presented a scenario in which 125 participants believed that they had completed a task and so had no reason to seek further solutions. To their surprise, after a period of distraction, we resumed the testing session. This novel method was combined with accurate recording of both response content and timing. The results from the second session a remarkable similarity n initial burst to those from the first, including a ideas, allowing the inference that, even in the absence of a reason to seek solutions, a process of nonconscious idea generation might be operating. PMID- 15362413 TI - An examination of the current usefulness of the Bem Sex-Role Inventory. AB - In the present study, an ethnically diverse convenience sample (N = 182; 62% female) of working adults (56%) and college students (M age=30.9 yr., in SD = 12.8, range= 18 to 71) completed the Bem Sex-Role Inventory which is a widely used self-report measure of perceptions of gender roles. Based on their scores, individuals' sex roles can be categorized as Masculine or Feminine (sex-typed) or Androgynous. The results of this study suggest that, almost 30 years after it was first developed, the categories can still be used to categorize men and women of varying ages. PMID- 15362414 TI - Mood states of formerly sedentary younger and older women at weeks 1 and 10 of a moderate exercise program. AB - This study assessed and contrasted mood changes associated with a 10-wk moderate exercise program in 64 formerly sedentary women of different ages. Analyses of covariance, with Week 1 mood scores as the covariates, indicated that the younger exercise group (n=32; M age=33 yr.) and older exercise group (n=32; M age=63 yr.) showed significantly (p<.05) more residual improvement in scores on McNair, Lorr, and Droppleman's Profile of Mood States Tension, Depression, Vigor, and Fatigue scales than a no-exercise control group (n=32; M age=48 yr.). There were no significant differences in amount of improvement between the younger and older exercise groups. Across both exercise groups, amount of mood score changes over 10 weeks were negatively correlated with initial mood scores (rs= -.22 to -.75). Limitations and research implications were discussed. PMID- 15362415 TI - An improved method for predicting which heavy drinkers become intoxicated. AB - Not all heavy drinkers become intoxicated. We sought to improve predictability of intoxication of heavy drinkers. Based on criteria for heavy drinking in the National Household Survey on Drug Abuse (NHSDA), we identified characteristics related to heavy drinking. We then created a dichotomous heavy drinker typology variable (yes/no) and determined how well we were able to identify drinkers who became intoxicated at least twice a month. Of those who fit this heavy drinking profile, 54% drank regularly to the point of intoxication, the same percent as those who become intoxicated in the self-reported heavy drinking group. However, 77% of those who fit both the profile and were self-described heavy drinkers, drank regularly to intoxication. We concluded that a demographic typology combined with self-reported drinking improves predictability of intoxication in heavy drinkers, and is a promising direction for research. PMID- 15362416 TI - Suicide Resilience Inventory-25: development and preliminary psychometric properties. AB - This manuscript describes the development of the Suicide Resilience Inventory-25, used to assess factors that help defend against suicidal thoughts and behaviors. We used multiple sources to generate and evaluate initial items (Study 1), then conducted an iterated principal-axis factor analysis with data from a combined sample of 540 adolescents and young adults. This identified three correlated factors, named Internal Protective, Emotional Stability, and External Protective. Estimated alpha for the total inventory and scales was high (.90 to .95). In Study 1, scores on the inventory significantly differentiated between the responses of adolescents and young adults, In Study 2, the inventory scores significantly differentiated between participants who reported (a) no prior suicide thoughts or attempts (145 men and 153 women), (b) brief suicidal thoughts (55 men and 110 women), and (c) prior suicide plans or attempts (22 men and 55 women). Results of the two studies suggest the inventory is useful for assessing the construct of suicide resilience in terms of these three operationally defined dimensions. PMID- 15362417 TI - Sensation seeking in a French population of pathological gamblers: comparison with regular and nongamblers. AB - To test the hypothesis that pathological gamblers in the French general population who play games available in cafes score lower on sensation seeking than regular and nongamblers, the Zuckerman's Sensation Seeking Scale was administered to 57 diagnosed pathological gamblers, 40 regular gamblers, and a control group of 97 nongamblers. Whereas pathological gamblers did not obtain a significantly lower Sensation Seeking mean than the regular and nongamblers groups, analysis indicated that those pathological gamblers seek different and multiple forms of games to reduce boredom, but they do not necessarily seek excitement-generating activities. One possible interpretation is that the types of games investigated in this study are "passive" forms of games that required low involvement from the gamblers. PMID- 15362418 TI - New frontiers for psychology and education: robotics. AB - The paper reviews the first attempts to study the educational and psychological usefulness of robotics: (1) the social and cooperative dimensions involved in the robot-building activities; (2) the reasoning strategies implied in building and programming robots; (3) the influences of robotics on mathematical and scientific achievement; (4) the use of robotics in modification of social skills of autistic children. PMID- 15362419 TI - Teachers' instructional efficacy and teachers' efficacy toward integration of information technologies in the classroom. AB - The aim of the study was to investigate the relationship between teachers' instructional efficacy and their efficacy toward integration of technologies in the classroom. A sample of 309 French Canadian elementary school teachers volunteered and were administered a French Canadian version of the Teacher Efficacy Scale and Teachers' efficacy scale toward integration of technologies in the classroom. Analysis yielded, as expected, a positive and significant partial correlation between the two types of self-efficacy beliefs (.27 and .36). PMID- 15362420 TI - Prayer and inner speech: is there a connection? AB - The present study explored the relation between frequency of prayer, inner speech, and self-efficacy for a sample of 134 Germany adults (88 women and 46 men). It was hypothesized that prayer activity by adults is functionally related to inner speech, and both might result in higher self-efficacy, but the findings challenge the notion that inner speech is associated with frequency of prayer and self-efficacy. More importantly, significant negative correlations were found between self-efficacy and both the measures of frequency of prayer (r = -.28) and church attendance (r = -.44). Further research is needed to relate inner speech to different types of private prayer, given that this sample comprised irregular churchgoers who prayed "never" or "sometimes." PMID- 15362421 TI - Comparing alcoholic and nonalcoholic parents on the family unpredictability scale. AB - Research findings and clinical observations suggest that families with an alcoholic parent are more unpredictable. Alcoholic parents (n=25, 68% men, 68% Euro-American, M age=38.6 yr.) and community parents (n=27, 52% men, 70% Euro American, M age=38.8 yr.) completed the self-report Family Unpredictability Scale of Ross and Hill. Alcoholic parents reported significantly higher (less predictability) scores on the subscales of Nurturance, Finances, and Discipline, as well as on the Total Family Unpredictability Scale (ps<.01). This appears to be the first study on family unpredictability and parental alcoholism in which parental reports of multiple dimensions of unpredictability are used. We provide suggestions for research and clinical uses of the scale, especially pertaining to families with an alcoholic parent. PMID- 15362422 TI - Reliability of a brief credit card attitude scale. AB - In a sample of 139 community residents, the three subscales from the brief Hayhoe, et al. credit card attitude scale were replicated in a factor analysis. PMID- 15362423 TI - Tolerance for error and computational estimation ability. AB - Previous investigators have suggested that the personality variable tolerance for error is related to success in computational estimation. However, this suggestion has not been tested directly. This study examined the relationship between performance on a computational estimation test and scores on the NEO-Five Factor Inventory, a measure of the Big Five personality traits, including Openness, an index of tolerance for ambiguity. Other variables included SAT-I Verbal and Mathematics scores and self-rated mathematics ability. Participants were 65 college students. There was no significant relationship between the tolerance variable and computational estimation performance. There was a modest negative relationship between Agreeableness and estimation performance. The skepticism associated with the negative pole of the Agreeableness dimension may be important to pursue in further understanding of estimation ability. PMID- 15362424 TI - The deadliness of mass murderers. AB - The death toll for mass murders was higher when the murderer committed suicide and when the incident was elsewhere than the Americas and Europe. PMID- 15362425 TI - Relationship of credit attitude and debt to self-esteem and locus of control in college-age consumers. AB - College-age consumers are one of the groups most highly targeted by credit card marketers. While some college students use their credit cards wisely, others are unable to control their spending. The objective of this study was to investigate differences in attitude toward credit cards and the psychological factors of self esteem and locus of control among college students who possess one or more credit cards. Attitude was operationalized to include three underlying components: cognitive, affective, and behavioral. We separated credit users into subcategories based on amount of installment debt. Convenience users were defined as those consumers who paid the credit-card balance in full each month. Installment users were classified as consumers who carried a balance month-to month. Convenience users were compared to mild and heavy installment users to assess significance of differences in attitudinal and psychological factors. There were no significant differences in the psychological factors across the credit-card user groups. In addition, there was a statistically significant difference on each of the attitude components (knowledge/beliefs, affect, and behavior) across user groups; convenience users, mild installment, and heavy installment users. PMID- 15362426 TI - Wilcoxon test after Levene's transformation can have an inflated Type I error rate. AB - It is shown that the procedure of applying the Wilcoxon test after Levene's transformation can have an inflated Type I error rate when distributions are skewed. Thus, when the data may come from an asymmetric distribution, the Wilcoxon test should not be applied as a test for homogeneity of variances after Levene's transformation. PMID- 15362427 TI - Cynicism about organizational change: an attribution process perspective. AB - The underlying attribution process for cynicism about organizational change is examined with six samples from four different organizations. The samples include hourly (n=777) and salaried employees (n= 155) from a manufacturing plant, faculty (n=293) and staff (n=302) from a large university, managers from a utility company (n=97), and young managers (n=65) from various organizations who were attending an evening MBA program. This form of cynicism is defined as the combination of Pessimism (about future change efforts) and a Dispositional attribution (why past efforts to change failed). Three analyses support this definition. First, an exploratory factor analysis (from the largest sample) produced two factors, one composed of Pessimism and the Dispositional attribution items and the second of the Situational attribution items. Second, the average correlation (across several samples) between Pessimism and Dispositional attribution is much higher (.59) than the average correlation between Pessimism and Situational attribution (.17). Third, scores on two different trait-based measures of cynicism correlate highest with the Dispositional attribution component of cynicism. A practical implication is that organizational leaders may minimize cynicism by managing both employees' pessimism about organizational change and employees' attributions about it. Specific suggestions for how this might be done are offered. PMID- 15362428 TI - Honesty when lighting votive candles in church: an informal look. AB - An informal enquiry conducted over 6 yr. suggested a decline from 92% to 28% of people paying for lighting votive candles in a large Catholic church. PMID- 15362429 TI - Relationships between five factor personality variables, workplace accidents, and self-efficacy. AB - Past research has examined the relationships between personality variables and workplace accidents; however, few studies have examined these relationships using the Five Factor Model of personality and trait-state relationships have not been examined in this context. The present study was undertaken to examine the relationships between personality characteristics based on the Five Factor Model, workplace accidents, and self-efficacy. To examine these relationships, personality, workplace accident, and self-efficacy data were collected from 202 undergraduate volunteers (134 women and 68 men) at a large midwestern urban university. The mean age of participants was 20.9 yr. (SD=5.0). The results indicated that scores on Agreeableness and Conscientiousness were significantly correlated with workplace accidents as were the self-efficacy variables. Neuroticism and Agreeableness were significantly related to self-efficacy. PMID- 15362430 TI - Clearing the supermarket shopping cart: an informal look. AB - An informal enquiry of the behavior of 500 supermarket shoppers clearing carts of litter prior to entering the store showed that 69% dumped the rubbish into another cart, 26% dropped it on the sidewalk, and 5% deposited it in a trash container. PMID- 15362431 TI - Convergent validity of the Beck Depression Inventory for Youth. AB - This study investigated the convergent validity of the Beck Depression Inventory for Youth in a sample of 34 children (22 boys, 12 girls) who ranged in age from 7 to 14 years (M= 11.3, SD=2.3) by comparing scores with simultaneous ratings on the Children's Depression Inventory. Analysis indicated that the sets of scores correlated very strongly regardless of age or sex. Given that this Beck Depression Inventory has fewer items and is guided by a strong theoretical premise, this scale may be a valuable tool for use with youth by clinicians in various mental health disciplines. PMID- 15362432 TI - Re-examining the role of interscholastic sport participation in education. AB - The purpose of the present study was to examine the relationships of interscholastic sports participation with locus of control, self-concept, cognitive test scores, and school dropout in adolescents via a secondary analysis of National Educational Longitudinal Study 1988 (NELS:88). Subjects were 1,883 students who completed all measurements in 8th grade (1988), at Follow-up 1 (10th, 1990), and Follow-up 2 (12th, 1992). Scales utilized to measure locus of control, self-concept, and cognitive ability were developed by the National Center for Educational Statistics. Dropout was assessed through school records and contact with participants. At baseline and Follow-up 1, students self reporting sport participation displayed higher self-concept and locus of control; however, these differences were no longer observed at Follow-up 2. Drop-out rate for boys at Follow-up 2 was significantly lower for students reporting interscholastic sport participation at baseline or at Follow-up 1. Similarly, drop-out rate for girls at Follow-up 2 was significantly lower for students reporting interscholastic sport participation at baseline or at Follow-up 1. These findings support the presumed positive role of sport participation for adolescents in American schools. PMID- 15362433 TI - The inner voice and the Plural Self. AB - In a sample of 182 undergraduates, scores on the Plural Self Scale were associated with scores on a scale designed to measure the presence of violent inner voices. PMID- 15362434 TI - Tolerable lifetime cadmium intake calculated from the inhabitants living in the Jinzu River basin, Japan. PMID- 15362435 TI - Arsenic exposure levels during cleanup of fly ash and dermatitis in an air sampling technician. PMID- 15362436 TI - Anomalous levels of heavy metals in sediments from Guaymas Bay, Mexico. PMID- 15362437 TI - Hydrogeological and hydrogeochemical features of the Heybeli Spa, Afyon, Turkey: arsenic and the other contaminants in the thermal waters. PMID- 15362438 TI - Arsenic bioaccumulation by beetles in an arsenic-rich region. PMID- 15362439 TI - Sequestration of nickel and copper by Azotobacter chroococcum SB1. PMID- 15362440 TI - Preliminary study on the application of municipal sludge to agriculture. PMID- 15362441 TI - Evaluation of an organoclay, an organoclay-anthracite blend, clinoptilolite, and hydroxy-apatite as sorbents for heavy metal removal from water. PMID- 15362442 TI - Effect of 2,4-dichlorophenoxyacetic acid on adsorption and release of copper on montmorillonite. PMID- 15362443 TI - Xylenes in Oregon hazelnuts. PMID- 15362444 TI - Study on health effects of disassembling imported electrical equipment. PMID- 15362445 TI - Acute toxicity assessment of 20 herbicides to the green alga Scenedesmus quadricauda (Turp.) Breb. PMID- 15362446 TI - Microbial reduction of weakly crystalline iron (III) oxides and suppression of methanogenesis in paddy soil. PMID- 15362447 TI - Finding hazardous waste sites: evaluation using the capture-recapture method. PMID- 15362448 TI - Evaluation of the Shk1 activated sludge bacterial luminescence inhibition assay: narcotic chemicals. PMID- 15362449 TI - Chlorinated pesticides and polychlorinated biphenyls in the coral reef skeleton of the Egyptian Red Sea coast. PMID- 15362450 TI - Distribution of organochlorine compounds in pine needles collected at urban sites in Croatia. PMID- 15362451 TI - Distribution of polycyclic aromatic hydrocarbons in Gomti river system, India. PMID- 15362452 TI - Trophic transfer efficiency of DDT to lake trout (Salvelinus namaycush) from their prey. PMID- 15362453 TI - Detection of phthalate metabolites in human amniotic fluid. PMID- 15362454 TI - Photosynthetic protein from Chlorella vulgaris strain Bt-09 may be responsible for the coping mechanism against cadmium toxicity. PMID- 15362455 TI - UVA light-induced toxic effects of 1-hydroxypyrene on human jurkat T-cells. PMID- 15362456 TI - Effects of cimetidine and phenobarbital on methylparathion metabolism in Hyalella azteca. PMID- 15362457 TI - Toxicokinetics of pyrene in tilapias Oreochromis niloticus following an intraperitoneal administration. PMID- 15362458 TI - Effect of water soluble fractions of diesel and an oil spill dispersant (Corexit 9527) on immune responses in mussels. PMID- 15362459 TI - Evaluation of ozone phytotoxicity in the greater area of a typical Mediterranean small city (Volos) and in the nearby forest (Pelion Mt.), central Greece. PMID- 15362460 TI - Acute toxic effects of neem-based insecticides on crustaceans. PMID- 15362461 TI - Toxicity of a synergized formulation of sumithrin to Daphnia magna. PMID- 15362462 TI - Effects of ammonia on the early life stages of northern pike (Esox lucius). PMID- 15362463 TI - Epidemiology of malaria presenting at British Columbia's Children's Hospital, 1984-2001: lessons for prevention. AB - BACKGROUND: Few studies have examined the epidemiology of imported malaria in Canadian children. Identifying populations at increased risk in Canada would enable targeted malaria prevention strategies within those groups. The study objective was to describe the epidemiology of malaria diagnosed at British Columbia's Children's Hospital (BCCH) between 1984 and 2001. METHODS: This was a retrospective chart review of malaria cases identified at BCCH and confirmed through the British Columbia Centre for Disease Control. Demographic and clinical data were recorded on a standardized form. RESULTS: Malaria was diagnosed 42 times in 40 children (age 24 days to 14.8 years). Thirty cases (71.4%) occurred in 28 Canadian residents, and 12 (28.6%) occurred in immigrant or refugee children. Twenty-six children (65%) were male. Thirty-one children (77.5%) were of East Indian descent. Thirty-three exposures (78.6%) to malaria occurred in the Indian subcontinent. Plasmodium vivax was identified in 37 cases (88.1%), P. falciparum in 3 (7.1%), and the species was unknown in 2 (4.8%). Fourteen cases in the resident children (46.7%) reported pre-travel counselling. Ten resident cases (33.3%) were prescribed chemoprophylaxis, primarily chloroquine, and at least six of them (60%) were non-compliant. The duration of symptoms prior to diagnosis was < 7 days in 27 cases (64.3%), 8 to 30 days in 10 (23.8%), > 30 days in 4 (9.5%) and the duration was unknown in 1 (2.5%). Twenty-four of 36 cases (66.7%) had seen 2 to more than 4 doctors before the diagnosis of malaria was made. CONCLUSION: The majority of children in our review were of East Indian origin and were exposed to malaria in India. Most had not sought or had received inadequate pre-travel counselling and had been non-compliant with chemoprophylaxis. As malaria is a potentially lethal but preventable disease, strategies to ensure adequate pre-travel counselling for high-risk groups are required. PMID- 15362464 TI - Pediatric tuberculosis in Alberta First Nations (1991-2000): outbreaks and the protective effect of bacille Calmette-Guerin (BCG) vaccine. AB - BACKGROUND: The tuberculosis control strategy of vaccinating First Nations newborns with BCG (bacille Calmette-Guerin) is currently undergoing re-evaluation in Canada. Review of recent pediatric tuberculosis morbidity could inform this re evaluation. METHODS: Potential source cases and pediatric cases of tuberculosis from Alberta First Nations were identified over the 10 years 1991-2000. The distribution of pediatric disease was described. The effect of BCG on tuberculosis morbidity in two large outbreaks was determined. RESULTS: A total of 57 potential source cases and 41 pediatric cases of tuberculosis were reported from 17 (41.5%) and 8 (19.5%) of the 41 on-reserve First Nation Community Health Centres, respectively. Three outbreaks traceable to three source cases accounted for 34 (18, 3, and 13, respectively) of the 41 (82.9%) pediatric cases. Each outbreak was spatially and temporally separate from the other. Each outbreak strain of Mycobacterium tuberculosis had a unique DNA fingerprint. In the largest outbreaks, disease-to-infection ratios (secondary case rates) were higher in newly infected unvaccinated versus vaccinated close pediatric contacts (12/13 [92.3%] versus 7/15 [46.7%], p=0.02), but the infection rate was almost certainly falsely high in the BCG vaccinated. One unvaccinated child had a brain tuberculoma in addition to primary pulmonary tuberculosis. CONCLUSION: For most Alberta First Nations communities, the spatial and temporal distribution of disease, and the meager impact on morbidity, challenge the rationale for continued use of BCG. PMID- 15362465 TI - Wearing masks in a pediatric hospital: developing practical guidelines. AB - During the outbreak of Severe Acute Respiratory Syndrome (SARS) in the spring of 2003, strict infection control measures were implemented in Toronto and surrounding hospitals. These measures included extreme restrictions on those who would normally accompany patients to the hospital, screening for SARS, and protective attire for hospital staff, including masks, face shields, goggles, gloves and gowns. At Toronto's Hospital for Sick Children (HSC), patients could only be accompanied or visited by one person, often only in patients' rooms. For the first four weeks, patients and their designated parent had to wear masks in almost all areas of the hospital. Staff wore masks (and other appropriate protective clothing) whenever in contact with patients and in many patient care areas. Although these barriers were an important part of containing SARS, their use created significant challenges for patients and staff. This article focusses on the use of infection control masks in routine pediatric healthcare and the tools developed by HSC staff to reduce the negative psychosocial impact on children and families. PMID- 15362466 TI - What factors are associated with poor developmental attainment in young Canadian children? AB - BACKGROUND: This study was undertaken to determine the association between poor developmental attainment (PDA) and biological, home environment and socio demographic factors in a population-based sample of Canadian children. METHODS: Cross-sectional data from two cycles (1994/95 and 1996/97) of the National Longitudinal Survey of Children and Youth were used. Children aged 1-5 years were included. PDA was defined as < or = 15th percentile for motor and social developmental skills (1-3 year olds) or Peabody Picture Vocabulary Test (4-5 year olds). Multiple logistic regression was used. RESULTS: The proportion of children with PDA varies across Canada, between males and females, and by age. Among 1 year olds in Cycle I, having a low birthweight (OR=3.3; 95% CI: 2.1-5.2), being male (OR=1.6; 95% CI: 1.2-2.2) and having a mother who is an immigrant (OR=1.6; 95% CI: 1.1-2.2) increased the odds of PDA. Similar results were observed in Cycle II. Among children aged 4-5 years in Cycle II, having a mother who is an immigrant (OR=5.3; 95% CI: 4.1-6.9) and a mother with low educational attainment (OR=2.8; 95% CI: 2.1-3.9) increased the odds of PDA. Low income was a significant predictor of PDA across all age groups. INTERPRETATION: The strong and consistent associations with living in a low-income household, having a mother with low educational attainment or a mother who is an immigrant highlight the need for targeting developmental assessments and services to this population. PMID- 15362467 TI - Dares to addiction: youth definitions and perspectives on gambling. AB - BACKGROUND: Over the past decade, there has been a rapid growth of gambling in Canada and internationally. Although youth are a potentially vulnerable group, little is known about what they understand and if they are being affected by the recent increase in gambling. METHODS: This study examined how youth view gambling using an inductive qualitative research design and analysis based on grounded theory principles. Twelve focus groups were conducted comprising 103 participants (median age = 15 years) with diverse representation of Ontario youth. Focus-group questions were designed to capture youth's experiences and opinions about gambling. RESULTS: Youth participants defined a spectrum of gambling from a dare and friendly betting to legalized forms of gambling (lotteries, casinos) and addiction. Their opinions varied according to age and gambling type. For example, daring and friendly betting were identified as positive activities used by younger adolescents to relieve boredom and establish social relationships. Gambling was separate from daring because of its association with money. Many participants had minimal awareness of the potential negative impact of gambling. Information technology (Internet) was seen as an attractive medium for playing games and gambling where no money was involved. CONCLUSION: Lack of awareness of gambling among youth and its consequences underscores the need for public education. The diverse range of gambling behaviour and age-dependent access to money need careful consideration in defining youth gambling "problems" and in designing public health interventions. PMID- 15362468 TI - Socio-demographic correlates of cigarette smoking among high school students: results from the British Columbia youth survey on smoking and health. AB - OBJECTIVE: To describe the association between selected socio-demographic factors (age, gender, ethnicity, and region) and the prevalence of smoking among adolescents in two regions of British Columbia, and to report recent findings related to the prevalence of tobacco use in British Columbia. METHODS: A cross sectional school-based survey was conducted using a random sample of 3,280 students from 13 schools in two regions of British Columbia. Multivariate logistic regression analysis was used to determine the association between age, region, gender and ethnicity and smoking status. RESULTS: Ethnic group membership was strongly associated with smoking status, which changed very little after controlling for the other socio-demographic factors. Controlling for age and ethnicity, the effect of gender on smoking status was moderated by region. CONCLUSION: Reliance on general population tobacco use prevalence rates masks important ethnic and gender differences. To increase the effectiveness of tobacco control policies and programming, greater attention needs to be paid to the socio demographic correlates associated with adolescents' tobacco use. PMID- 15362469 TI - Evaluation of health care needs and patient satisfaction among hepatitis C patients treated at a hospital-based, viral hepatitis clinic. AB - OBJECTIVE: To define self-reported hepatitis C knowledge, health care needs, and patient satisfaction in a representative cohort of hepatitis C virus (HCV) infected adults treated at a university hospital-based viral hepatitis clinic in Canada. METHODS: A questionnaire package evaluating HCV knowledge, health care needs, and patient satisfaction was administered to 111 consecutive consenting HCV patients during their first and 10-month follow-up HCV clinic visits. RESULTS: At their first HCV clinic visit, 52% of patients rated their current HCV knowledge as "fair" or "poor". Patients identified HCV education, quality medical care, medication coverage, and psychological counselling as important HCV health care needs. Health care satisfaction outcome data at 10-month follow-up indicated that patients felt significantly better informed, more satisfied, and more actively involved in their HCV health care. CONCLUSION: A bio-psychosocial framework in which medical, psychological, educational, and social issues are addressed is desirable for optimal HCV health care. PMID- 15362470 TI - Smoking cessation for pregnant women: current Canadian programs and future development. PMID- 15362471 TI - Consultation on strengthening the Pan-Canadian public health system and meeting with the Minister of State (public health). PMID- 15362472 TI - [The social representation of pregnancy in adolescence. A Quebec study. ]. AB - OBJECTIVE: To gather information on the themes underlying social representations of teenage pregnancy among adolescent girls and boys and people working with them (health/social/school workers). METHOD: A qualitative approach was used. Adolescents and health/social/school workers were gathered separately in focus groups. A content analysis of their discourses was done. RESULTS: This study shows that adolescents and health/social/school workers were not homogeneous groups. Their representations on teenage pregnancy varied and were sometimes contradictory. Several points of view were expressed that gave rise to four distinct positions: negative, positive, ambivalent and dynamic. It is from these four positions that four social representations of teenage pregnancy were defined: pregnancy as a problem, pregnancy as a project, pregnancy as a tension, pregnancy as a gain of power. Within these four representations, the discourses were divided into four dimensions: emotional, introspective, psychobiological and socio-economic. DISCUSSION: The similarities and divergences between the discourses of adolescents and those of health/social/school workers reveal that the representations of teenage pregnancy are neither preconceived nor unalterable. They have functional properties of adaptation to changing realities, individual conditions, ongoing events, times of crisis and diverse social and cultural grounds. This study presents a new perspective on the topic of teenage pregnancy and questions people's attitude and relationship with teenagers with respect to this issue. PMID- 15362473 TI - Evaluation of a cervical cancer screening intervention for prison inmates. AB - BACKGROUND: Female prison inmates are underscreened and are at higher risk of cervical cancer. The impact of a nurse-led Pap screening intervention was examined, which included information sessions and Pap testing clinics. METHOD: Pap screening rates for 650 inmates at the Burnaby Correctional Centre for Women were compared both before and during the 20-week intervention period. These rates were determined by record linkage of Correction Branch inmate records and Cervical Cancer Screening Program patient records. Associations between socio demographic factors and Pap screening rates were also examined. RESULTS: A higher proportion of inmates was screened during the intervention period (26.9%) than during the preintervention period (21.0%) (although the difference was not statistically significant (p=0.06)). Very short-stay inmates were less frequently screened in the preceding two years before the intervention. Inmates with no high school education and longer lengths of incarceration were significantly more likely to receive Pap testing during the intervention period as compared to the preintervention period. CONCLUSION: The nurse-led intervention resulted in a modest improvement in the proportion of inmates receiving Pap screening. Unfortunately, the benefit of the nurse clinician did not reach, to a greater extent, inmates who had not been previously screened or who were inadequately screened. There is need for further work to target this hardest-to-reach group. PMID- 15362474 TI - Individual and regional determinants of mammography uptake. AB - BACKGROUND: Analysis of mammography utilization has traditionally been performed from an individual-level perspective. The purpose of this study was to explore the combined influence of individual- and regional-level determinants of mammography utilization. METHODS: Logistic hierarchical multilevel modelling was used to investigate the influences of region of residence and individual characteristics on mammography utilization. Socioeconomic status information about health planning regions was derived from the 1996 Canadian Census. Individual-level information was extracted from the 1996 National Population Health Survey. RESULTS: After controlling for individual-level education, regions with fewer high school graduates had lower levels of mammography utilization. A cross-level interaction between regional-level education and individual-level social involvement was found. Other individual-level variables associated with screening confirmed previous literature findings. CONCLUSION: Our findings suggest that higher levels of participation in social activities modify the detrimental influence on mammography utilization of living in a less educated region. This challenges the current focus of mammography screening research on individual-level determinants of uptake. Multilevel, synergistic strategies to possibly achieve higher levels of screening should be considered by health promotion program planners. PMID- 15362475 TI - Evaluation of a re-training program for older drivers. AB - BACKGROUND: Some older drivers may have a higher crash risk than others. Because many of these drivers have not received formal driving training, we evaluated the effectiveness of a re-training program for older drivers as it is currently being offered. Specifically, we examined if older drivers who received the training would have better scores on a driving evaluation than drivers who did not receive the training. METHODS: We used a randomized controlled trial. Participants first took part in on-road driving evaluations (possible scores ranged from 0 to 100), after which they were block randomized into training (treatment) or waiting (control) groups based on the driving evaluation results, age, and sex. Individuals in the treatment group attended driver re-training sessions, and were tested once more with the on-road test. Those in the control group completed their second driving evaluation and were then offered the re-training sessions. The second evaluations took place approximately two months after the first evaluations. RESULTS: Sixty-five participants completed two driving evaluations. Despite the overall improvement in driving scores (3.73, SD = 6.87, p = 0.001), we found no statistically significant difference between the control and treatment groups. The mean improvement for the control group was 3.46 (SD=6.72) compared to 4.02 (SD=7.11) for the treatment group (p = 0.747). Drivers' age was related to overall driving scores (r = -0.55, p = 0.001) but not changes between the first and second evaluations (r = 0.01, p = 0.955). INTERPRETATION: Although we have not demonstrated a statistically significant impact of the intervention, the overall increase suggests that an initial driving evaluation may underestimate the actual driving ability of many older drivers. Furthermore, although older drivers may have lower driving scores initially, they have the ability to improve on these scores. These findings should encourage us to explore diverse approaches to improve driving safety. PMID- 15362476 TI - What do Canadian seniors say supports their quality of life? Findings from a national participatory research study. AB - BACKGROUND: A national project investigated seniors' perceptions of the influences upon their quality of life. The seven participating cities were Montreal, Quebec City, Ottawa, Toronto, Regina, Vancouver and Whitehorse. The project focussed on policy decisions affecting the quality of life of seniors. It was a participatory study in which seniors controlled the direction and shape of the project in each city. METHODS: Focus groups and individual interviews with seniors and stakeholders. Data analysis used qualitative methods to see the world through the eyes of participants. Each project was committed to hearing the voices of seniors and their views on which issues were affecting the quality of their lives. RESULTS: Across the seven cities, seniors highlighted access to information, health care, housing, income security, safety and security, social contacts and networks, and transportation as key issues that affect the quality of life of seniors in Canada. CONCLUSIONS: The findings affirm the value of participatory activities that involve seniors working with other sectors as a productive policy-informing approach. The Seniors' Quality of Life projects demonstrate the conceptual power of the determinants of health perspective to understand seniors' quality of life issues. While seniors considered health care to be a continuing concern, they also recognized socio-economic issues as significantly affecting the quality of their lives. PMID- 15362477 TI - Beyond access: who reports that health care is not being received when needed in a publicly-funded health care system? AB - OBJECTIVES: To examine the relationship between reporting that health care was not received when needed in Canada's publicly-funded health care system and contact with the health care system, and to explore whether there is a differential impact of specific chronic conditions. METHODS: Analyses were carried out on the 2000/2001 Canadian Community Health Survey. The proportion of survey respondents who reported that they had not received health care when it was needed was determined overall, and for individuals with selected socio demographic, health status, and health care utilization characteristics, as well as stratified by various chronic conditions. Multivariate logistic regression was then employed to examine possible predictors of having reported that health care was not received when it was needed during the previous year. RESULTS: Approximately 13% of Canadians reported that health care was not received when they felt it was needed during the previous year. These individuals reported higher rates of health care utilization, worse health status, were more likely to have chronic health conditions (particularly conditions such as fibromyalgia, Crohn's disease, and chronic bronchitis for which there is no effective treatment), and were more likely to be female, younger, white, have higher education and lower income. CONCLUSION: Reporting that health care was not received when it was needed may not be related to accessibility but rather to a perceived failure of the system to meet the individual's needs. PMID- 15362478 TI - From stool to statistics: reporting of acute gastrointestinal illnesses in Canada. AB - BACKGROUND: Limitations associated with the under-reporting of enteric illnesses have long been recognized but the extent and variation of this under-reporting in Canada has not been examined. Given the public health value of surveillance data, a closer examination of under-reporting of enteric illnesses in Canada was warranted. METHODS: Paper-based surveys were administered (a) to all laboratories in Canada licensed to process stool specimens and (b) to all local public health authorities in two provinces. RESULTS: Of the laboratories surveyed, 67% (n=274) conducted on-site testing of stool specimens for enteric bacteria, 31% (n=126) for parasites and 10% (n=42) for viruses. In the year 2000, these laboratories processed 459,982 stool specimens, of which 5%, 15%, 8% and 19% were positive for enteric bacteria (excluding C. difficile), C. difficile, parasites and viruses, respectively. Variations in laboratory testing and health authority reporting protocols and policies were identified. Of the laboratory-confirmed cases of AGI reported to local public health authorities, 5% (n=846) were not reported to provincial counterparts. CONCLUSION: A significant proportion of AGI cases submitting stool specimens are not captured in Canada's passive surveillance system due to unknown etiology. A much smaller proportion of laboratory-confirmed cases reported to local public health authorities are not captured at the provincial or national level. Given that the number of laboratory-confirmed AGI cases represents such a small fraction of all community cases, strategies to compensate for under-reporting and efforts directed at harmonizing laboratory and local public health authority policies and practices would be welcomed. PMID- 15362479 TI - Validity of using surname to define Chinese ethnicity. PMID- 15362480 TI - Alterations in atrial natriuretic peptide and its receptors in streptozotocin induced diabetic rat kidneys. AB - In this study the effect of diabetes mellitus on atrial natriuretic peptide (ANP) receptors in streptozotocin- (STZ-) induced diabetic rat kidneys was studied. Moreover, plasma ANP concentration was evaluated in diabetic and control rats by using radioimmunoassay. In addition, the expression of ANP in the kidneys of control and diabetic rats was evaluated by immunohistochemistry. Body-weight loss and increased glucose levels were used as indices of diabetes mellitus in the STZ induced rats. There was a significant loss in the body weight of the diabetic rats compared to controls. The efficacy of STZ administration was confirmed by rising blood glucose levels, which were significantly higher in diabetic rats compared to controls. Plasma ANP concentration was significantly greater in the diabetic rats in comparison with controls. Moreover, our immunohistochemical results show that the expression of ANP in diabetic rats was higher than that in age-matched controls. ANP was observed in the cells lining the proximal convoluted tubules in the cortex. The distribution and levels of ANP receptors in the kidneys of diabetic rats and age-matched controls were investigated using quantitative receptor autoradiography. Our results demonstrate significant decrease in ANP receptors in the kidneys of the diabetic rats compared to controls. The significant decrease was found in the juxtaglomerular medulla, inner medulla, and the papillae. The decrease in ANP receptors observed in the diabetic kidneys could have pathological consequences resulting in renal resistance to ANP in diabetes. PMID- 15362482 TI - Fructose transport and metabolism in adipose tissue of Zucker rats: diminished GLUT5 activity during obesity and insulin resistance. AB - Fructose is a major dietary sugar, which is elevated in the serum of diabetic humans, and is associated with metabolic syndromes important in the pathogenesis of diabetic complications. The facilitative fructose transporter, GLUT5, is expressed in insulin-sensitive tissues (skeletal muscle and adipocytes) of humans and rodents, where it mediates the uptake of substantial quantities of dietary fructose, but little is known about its regulation. We found that GLUT5 abundance and activity were compromised severely during obesity and insulin resistance in Zucker rat adipocytes. Adipocytes from young obese (fa/fa), highly insulin responsive Zucker rats contained considerably more plasma membrane GLUT5 than those from their lean counterparts (1.8-fold per microgram membrane protein), and consequently exhibited higher fructose transport (fivefold) and metabolism (threefold) rates. Lactate production was the preferred route for fructose metabolism in these cells. As the rats aged and become more obese and insulin resistant, adipocyte GLUT5 surface density (12-fold) and fructose transport (10 fold) and utilisation rates (threefold) fell markedly. The GLUT5 loss was more dramatic in adipocytes from obese animals, which developed a more marked insulin resistance than lean counterparts. The decline of GLUT5 levels in adipocytes from older, obese animals was not a generalised effect, and was not observed in kidney, nor was this expression pattern shared by the alpha1 subunit of the Na+/K+ ATPase. Our findings suggest that plasma membrane GLUT5 levels and thus fructose utilisation rates in adipocytes are dependent upon cellular insulin sensitivity, inferring a possible role for GLUT5 in the elevated circulating fructose observed during diabetes, and associated pathological complications. PMID- 15362481 TI - Differential effects of vanadium, tungsten and molybdenum on inhibition of glucose formation in renal tubules and hepatocytes of control and diabetic rabbits: beneficial action of melatonin and N-acetylcysteine. AB - Effect of vanadyl acetylacetonate (VAc), tungstate and molybdate on gluconeogenesis has been studied in isolated hepatocytes and kidney-cortex tubules. In renal tubules of control and alloxan-diabetic animals, the rank order of the metal-compounds-induced (i) inhibition of glucose formation from alanine+glycerol+octanoate or aspartate+glycerol+octanoate, (ii) decrease in the mitochondrial membrane potential (delta psim), (iii) increase in the hydroxyl free radicals (HFR) generation and (iv) decline in glucose-6-phosphatase activity was the following: VAc > tungstate > molybdate. Moreover, in contrast to VAc, both tungstate and molybdate at 100 microM concentration did not practically decrease glucose production in hepatocytes isolated from diabetic rabbits, and significantly increased the rate of lactate formation in renal tubules. N acetylcysteine at 2 mM concentration partially attenuated vanadium-induced alterations in glucose formation, delta psim and the cellular glutathione redox state, whereas 0.1 mM melatonin did not abolish vanadium-induced changes in gluconeogenesis despite attenuation of vanadium effects on HFR formation and delta psim decline. However, similarly to control rabbits, following 6 days of intraperitoneal administration of both VAc (1.275 mg V/kg body weight daily) and melatonin (1 mg/kg body weight daily) to alloxan-diabetic animals, vanadium induced elevated serum creatinine and urea levels were decreased, indicating the beneficial effect of melatonin on diabetes- and vanadium-induced nephrotoxicity in rabbits. As serum glucose levels were also significantly diminished by vanadium+melatonin treatment of diabetic animals, the combination therapy of vanadium compounds and melatonin needs a careful evaluation. PMID- 15362483 TI - Beneficial effect of vitamin E on the metabolic parameters of diabetic rats. AB - The role of vitamin E in the pathogenesis of diabetes mellitus is unknown. The purpose of this study was to examine the effect of oral administration of vitamin E on some of the metabolic parameters of experimental diabetic rats. Diabetes was induced by intraperitoneal injection of streptozotocin (60 mg/kg body weight at 12 weeks of age). Vitamin E (0.2, 0.4, 0.8 mg/kg body weight) was administered orally for a period of 3 weeks to normal and diabetic Wistar rats. In some experiments, Vitamin E was given either before or after the induction of diabetes mellitus. Blood glucose level and weight were recorded for each rat in different groups on a weekly basis. Oral glucose tolerance test (OGTT) was performed on fasted normal, diabetic and vitamin E treated rats at the end of the experiment. Vitamin E significantly (p < 0.01) reduced blood glucose levels in experimental diabetes mellitus at all doses as compared to untreated rats. Vitamin E induced weight loss in normal as well as in diabetic rats. The beneficial effect of vitamin E on the hyperglycaemia of diabetic rats was dose-dependent. Moreover, vitamin E also improved OGTT in diabetic rats compared to untreated diabetics. In conclusion, vitamin E may play a role in glucose metabolism and thus be a useful adjuvant therapy in type I diabetes. PMID- 15362484 TI - Effect of maternal diabetes and ethanol interactions on embryo development in the mouse. AB - The aim of this study was to determine the possible fetal effects of interaction between maternal diabetes and acute doses of alcohol. Pregnant TO mice were made diabetic by a single injection of streptozotocin (STZ) on gestation day (GD) 2. Single dose of 0.003 or 0.03 ml/g body weight of fresh ethanol (25% v/v of absolute alcohol in normal saline) was injected into groups of diabetic and nondiabetic animals on GD 7 or 8. One group of diabetic animals had a daily dose of 6-8 IU of insulin subcutaneously. Fetuses were collected on GD 18. There was a significant increase in the incidence of implantation failure in the diabetes plus ethanol groups and insulin control group. Ethanol injection on GD 7 accentuated diabetes-related embryonic resorption and intrauterine growth retardation (IUGR). This effect was less marked in the diabetic group treated with ethanol on GD 8. Diabetes alone produced a greater incidence of IUGR than ethanol alone. Midfacial hypoplasia and minor anomalies were found more frequently in the combination treatment groups. Holoprosencephaly and thymus hypoplasia observed in diabetic groups were found to be reduced in frequency in the diabetes plus ethanol groups, suggesting an antagonistic type of ethanol diabetes interaction, stage-dependently. Since severely malformed embryos are known to be resorbed/killed in utero in mice, this reduction might reflect the magnitude of early death of severely malformed embryos. These data suggest that the interaction effects are possibly related to alterations in fundamental developmental processes of early embryos. PMID- 15362485 TI - Orally administered tryptophan and experimental type 2 diabetes. AB - There is a link between diabetes and oxidative stress. Hyperglycaemia leads to free radical generation and alterations of endogenous antioxidants. Our aim is to study the effect of orally administered L-tryptophan (TRP), the melatonin precursor, an endogenous antioxidant, on circulating levels of glycaemia, insulin and melatonin, and on the superoxide dismutase and catalase antioxidant systems in non-diabetic (ND) and type 2 diabetic (n5-STZ) male Wistar rats. At 19:30 every day for 15 days, TRP (125 mg/kg body weight) was administered orally. At 09:00 every two days the glycaemia was measured and every day the intake of food and water was recorded. At the beginning and end of treatment (at 09:00; 21:00; 02:00) plasma insulin and melatonin levels were measured, and (at 09:00) the enzymatic activities of catalase and superoxide dismutase (SOD) in erythrocytes were also measured. Glycaemia values were greater (p < 0.01) in n5-STZ rats than in ND rats, while insulin levels were lower (p < 0.05) at all times studied and these parameters were not altered by the TRP administration. Melatonin levels at 02:00 were lower in n5-STZ than in ND rats (p < 0.05). The TRP administration did not modify the circulating melatonin levels in ND rats, but raised (p < 0.01) the levels at 02:00 in the treated n5-STZ group. In ND rats after TRP administration there was a decline in catalase activity (p < 0.05), while in n5-STZ rats there was a rise (p < 0.01) at the end of treatment. However, there were no significant changes in SOD activity. There was increased food intake (g/day) in the treated n5-STZ group (p < 0.01). In conclusion, the oral administration of TRP did not modify glycaemia or insulinaemia levels, but raised melatonin levels in diabetic rats at 02:00, lowered catalase activity in ND rats but raised it in n5-STZ rats, and increased food intake in n5-STZ rats. PMID- 15362486 TI - Beneficial effects and mechanism of action of Momordica charantia juice in the treatment of streptozotocin-induced diabetes mellitus in rat. AB - This study investigated the beneficial effects and mechanism of action of the juice of Momordica charantia in streptozotocin (STZ)-induced diabetes mellitus in rats. Diabetes mellitus was associated with significant (p < 0.01) time course reductions in body weight, plasma insulin and the number of insulin positive cells per islet and significant (p < 0.01) time course elevation in blood glucose and osmolarity and systolic blood pressure compared to age-matched healthy controls. Oral intake of M. charantia juice by STZ-induced diabetic rats partially reversed all the diabetes-induced effects measured. Daily oral administration of M. charantia juice to STZ-induced diabetic rates significantly (p < 0.01) reduced the Na+- and K+-dependent absorptions of glucose by the brush border membrane vesicles of the jejunum compared to the responses obtained in STZ induced diabetic rat. Either insulin (100 MM) or the fruit juice lyophilised extract (5 microg x ml(-1)) can stimulate 14C-D-glucose uptake in L6 myotubes. These effects were completely blocked by wortmannin, an inhibitor of phosphatidylinositol 3-kinase. High concentrations (10-200 microg x ml(-1)) of M. charantia juice extract inhibited 14C-D-glucose uptake in L6 myotubes compared to the control response. The effect of M. charantia treatment was also investigated on myelinated fibre abnormalities in the tibial nerve of STZ-induced diabetic and control rats. The results show that diabetes was associated with significant (p < 0.05) reduction in the mean cross-sectional myelinated nerve fibres, axonal area, myelin area and maximal fibre area compared to end controls. Treatment of STZ induced diabetic rats with M. charantia juice normalised the structural abnormalities of peripheral nerves. The results indicate that M. charantia can exert marked beneficial effects in diabetic rats, and moreover, it can regulate glucose uptake into jejunum membrane brush border vesicles and stimulate glucose uptake into skeletal muscle cells similar to the response obtained with insulin. PMID- 15362487 TI - Comparison of raloxifene and atorvastatin effects on serum lipids composition of healthy post-menopausal women. AB - The aim of this study was to evaluate the effects of the selective oestrogen receptor modulator, raloxifene, and those of statin, atorvastatin, in reducing the cardiovascular risks associated with the post-menopausal status. A detailed study of serum lipid concentrations was performed in four groups of post menopausal women receiving either placebo, raloxifene or atorvastatin alone or their combination for the period of three months. Group A (raloxifene) showed significant decrease in total cholesterol levels (P < 0.05) and an increase in phospholipids concentration (P < 0.05), followed by a marked reduction in low density lipoprotein cholesterol (LDL-C) levels (P < 0.01) and ApoB amounts (P < 0.001). Additionally, ApoA-I concentration was significantly increased (P < 0.01). Group B (atorvastatin) presented decreased cholesterol (P < 0.05) and triglycerides levels (P < 0.01), followed by elevated high-density lipoprotein cholesterol (HDL-C) concentration (P < 0.05) and low LDL-C amounts (P < 0.001). ApoA-I was significantly increased (P < 0.001) whereas ApoB was reduced (P < 0.001). The combined treatment in Group C (raloxifene and atorvastatin) showed significant changes in the majority of serum lipids. In particular, total cholesterol was reduced (P < 0.001), as well as triglycerides (P < 0.001) levels. Phospholipids were raised (P < 0.01) whereas LDL-C was reduced (P < 0.001) as was ApoB (P < 0.001). Furthermore, ApoA-I was elevated (P < 0.001). A further attempt to evaluate each treatment group was performed and the significance of these results is discussed. PMID- 15362488 TI - Effects of age and streptozotocin-induced diabetes on biogenic amines in rat tail artery. AB - The changes in amine concentrations in different segments of the rat tail artery have been investigated at different ages and after different durations of streptozotocin-induced diabetes. There was a significant positive slope to the relationship between amine concentrations and age in proximal portion of the normal tail artery, and a significant additional increase in amine concentrations following induction of diabetes. The peak of the latter response occurred between 10 and 20 weeks following the induction of diabetes. There was also a significant dependence on the length of the post-ganglionic neurones, which may be related to the failure of axonal transport of some of the essential enzymes or transporters for these biogenic amines. This model of altered catecholamine metabolism and handling requires further investigation so that alterations in the mechanisms involved in processing of these amines in diabetic autonomic neuropathy may be elucidated. PMID- 15362489 TI - Mechanism of exocrine pancreatic insufficiency in streptozotocin-induced diabetes mellitus in rat: effect of cholecystokinin-octapeptide. AB - This study investigated the effects of cholecystokinin-octapeptide (CCK-8) on pancreatic juice flow and its contents, and on cytosolic calcium (Ca2+) and magnesium (Mg2+) levels in streptozotocin (STZ)-induced diabetic rats compared to healthy age-matched controls. Animals were rendered diabetic by a single injection of STZ (60 mg kg(-1), I.P.). Age-matched control rats obtained an equivalent volume of citrate buffer. Seven weeks later, animals were either anaesthetised (1 g kg(-1) urethane; IP) for the measurement of pancreatic juice flow or humanely killed and the pancreas isolated for the measurements of cytosolic Ca2+ and Mg2+ levels. Non-fasting blood glucose levels in control and diabetic rats were 92.40 +/- 2.42 mg dl(-1) (n = 44) and >500 mg dl(-1) (n = 27), respectively. Resting (basal) pancreatic juice flow in control and diabetic anaesthetised rats was 0.56 +/- 0.05 ul min(-1) (n = 10) and 1.28 +/- 0.16 ul min(-1) (n = 8). CCK-8 infusion resulted in a significant (p < 0.05) increase in pancreatic juice flow in control animals compared to a much larger increase in diabetic rats. In contrast, CCK-8 evoked significant (p < 0.05) increases in protein output and amylase secretion in control rats compared to much reduced responses in diabetic animals. Basal [Ca2+]i in control and diabetic fura-2 loaded acinar cells was 109.40 +/- 15.41 nM (n = 15) and 130.62 +/- 17.66 nM (n = 8), respectively. CCK-8 (10(-8)M) induced a peak response of 436.55 +/- 36.54 nM (n = 15) and 409.31 +/- 34.64 nM (n = 8) in control and diabetic cells, respectively. Basal [Mg2+]i in control and diabetic magfura-2-loaded acinar cells was 0.96 +/- 0.06 nM (n = 18) and 0.86 +/- 0.04 nM (n = 10). In the presence of CCK-8 (10(-8)) [Mg2+]i in control and diabetic cells was 0.80 +/- 0.05 nM (n = 18) and 0.60 +/- 0.02 nM (n = 10), respectively. The results indicate that diabetes-induced pancreatic insufficiency may be associated with derangements in cellular Ca2+ and Mg2+ homeostasis. PMID- 15362490 TI - Diabetes-induced changes in glucose synthesis, intracellular glutathione status and hydroxyl free radical generation in rabbit kidney-cortex tubules. AB - Diabetes-induced changes in glucose formation, intracellular and mitochondrial glutathione redox states as well as hydroxyl free radicals (HFR) generation have been investigated in rabbit kidney-cortex tubules. In contrast to renal tubules of control animals, diabetes-evoked increase in glucose formation in the presence of either aspartate+glycerol+octanoate or malate as gluconeogenic precursors (for about 50%) was accompanied by a diminished intracellular glutathione reduced form (GSH)/glutathione oxidised one (GSSG) ratio by about 30-40%, while the mitochondrial GSH/GSSG ratio was not altered. However, a relationship between the rate of gluconeogenesis and the intracellular glutathione redox state was maintained in renal tubules of both control and diabetic rabbits, as concluded from measurements in the presence of various gluconeogenic precursors. Moreover, diabetes resulted in both elevation of the glutathione reductase activity in rabbit kidney-cortex and acceleration of renal HFR generation (by about 2-fold). On the addition of melatonin, the hormone exhibiting antioxidative properties, the control values of HFR production were restored, suggesting that this compound might be beneficial during diabetes therapy. In view of the data, it seems likely that diabetes-induced increase in HFR formation in renal tubules might be responsible for a diminished intracellular glutathione redox state despite elevated glutathione reductase activity and accelerated rate of gluconeogenesis, providing glucose-6-phosphate for NADPH generation via pentose phosphate pathway. PMID- 15362491 TI - Momordica charantia fruit juice stimulates glucose and amino acid uptakes in L6 myotubes. AB - The fruit of Momordica charantia (family: Cucurbitacea) is used widely as a hypoglycaemic agent to treat diabetes mellitus (DM). The mechanism of the hypoglycaemic action of M. charantia in vitro is not fully understood. This study investigated the effect of M. charantia juice on either 3H-2-deoxyglucose or N methyl-amino-a-isobutyric acid (14C-Me-AIB) uptake in L6 rat muscle cells cultured to the myotube stage. The fresh juice was centrifuged at 5000 rpm and the supernatant lyophilised. L6 myotubes were incubated with either insulin (100 nM), different concentrations (1-10 microg ml(-1)) of the juice or its chloroform extract or wortmannin (100 nM) over a period of 1- 6 h. The results were expressed as pmol min(-1) (mg cell protein)(-1), n = 6-8 for each value. Basal 3H deoxyglucose and 14C-Me-AIB uptakes by L6 myotubes after 1 h of incubation were (means +/- S.E.M.) 32.14 +/- 1.34 and 13.48 +/- 1.86 pmol min(-1) (mg cell protein)(-1), respectively. Incubation of L6 myotubes with 100 nM insulin for 1 h resulted in significant (ANOVA, p < 0.05) increases in 3H-deoxyglucose and 14C-Me AIB uptakes. Typically, 3H-deoxyglucose and 14C-Me-AIB uptakes in the presence of insulin were 58.57 +/- 4.49 and 29.52 +/- 3.41 pmol min(-1) (mg cell protein( 1)), respectively. Incubation of L6 myotubes with three different concentrations (1, 5 and 10 microg ml(-1)) of either the lyophilised juice or its chloroform extract resulted in time-dependent increases in 3H-deoxy-D-glucose and 14C-Me-AIB uptakes, with maximal uptakes occurring at a concentration of 5 microg ml(-1). Incubation of either insulin or the juice in the presence of wortmannin (a phosphatidylinositol 3-kinase inhibitor) resulted in a marked inhibition of 3H deoxyglucose by L6 myotubes compared to the uptake obtained with either insulin or the juice alone. The results indicate that M. charantia fruit juice acts like insulin to exert its hypoglycaemic effect and moreover, it can stimulate amino acid uptake into skeletal muscle cells just like insulin. PMID- 15362492 TI - Effect of insulin on exocrine pancreatic secretion in healthy and diabetic anaesthetised rats. AB - This investigation characterised the effects of exogenous insulin on exocrine pancreatic secretion in anaesthetised healthy and diabetic rats. Animals were rendered diabetic by a single injection of streptozotocin (STZ, 60 mg kg(-1) I.P.). Age-matched controls were injected citrate buffer. Rats were tested for hyperglycaemia 4 days after STZ injection and 7-8 weeks later when they were used for the experiments. Following anaesthesia (1 g kg(-1) urethane I.P.), laparotomy was performed and the pancreatic duct cannulated for collection of pure pancreatic juice. Basal pancreatic juice flow rate in diabetic rats was significantly (p < 0.001) increased whereas protein and amylase outputs were significantly (p < 0.001) decreased compared to control rats. Insulin (1 IU, I.P.) produced in healthy rats significant increases in pancreatic flow rate, amylase secretion and protein output compared to basal (p < 0.05). Insulin action also included a reduction in blood glucose (152.7 +/- 16.9 mg dl(-1), n = 6, prior to insulin and 42.0 +/- 8.4 mg dl(-1), n = 4, 100 min later). In fact, flow rate and glycaemia showed a strong negative correlation (p < 0.01, Pearson). Pretreatment with atropine (0.2 mg kg(-1), I.V.) abolished the effects of insulin on secretory parameters despite a similar reduction in glycaemia; in this series of experiments the correlation between flow rate and blood glucose was lost. In diabetic rats, insulin (4 IU, I.P.) did not modify exocrine pancreatic secretion. There was a fall in blood glucose (467.6 +/- 14.0 mg dl(-1), n = 10, prior to insulin and 386.6 +/- 43.6 mg dl(-1), n = 7, 120 min later). Rats, however, did not become hypoglycaemic. Similar results were observed in diabetic atropinized rats. The results of this study indicate that the effects of insulin on exocrine pancreatic secretion in anaesthetised healthy rats are mediated by hypoglycaemia evoked vagal cholinergic activation. PMID- 15362493 TI - Concentrations of signal transduction proteins exercise and insulin responses in rat extensor digitorum longus and soleus muscles. AB - Differences in the concentrations of signal transduction proteins often alter cellular function and phenotype, as is evident from numerous, heterozygous knockout mouse models for signal transduction proteins. Here, we measured signal transduction proteins involved in the adaptation to exercise and insulin signalling in fast rat extensor digitorum longus (EDL; 3% type I fibres) and the slow soleus muscles (84% type I fibres). The EDL and soleus were excised from four rats, the proteins extracted and subjected to Western blots for various signal transduction proteins. Our results show major differences in signal transduction protein concentrations between EDL and soleus. The EDL to soleus concentration ratios were: Calcineurin: 1.43 +/- 0.10; ERK1: 0.38 +/- 0.18; ERK2: 0.61 +/- 0.16; p38alpha, beta: 1.36 +/- 0.15; p38gamma/ERK6: 0.95 +/- 0.11; PKB/AKT: 1.44 +/- 0.08; p70S6k: 6.86 +/- 3.58; GSK3beta: 0.69 +/- 0.03; myostatin: 1.95 +/- 0.43; NF-kappaB: 0.32 +/- 0.10 (values >1 indicate higher expression in the EDL, and values < 1 indicate higher expression in the soleus). With the exception of p38gamma/ERK6, the concentration of each signal transduction protein was uniformly higher in one muscle than in the other in all four animals. These experiments show that signal transduction protein concentrations vary between fast and slow muscles, presumably reflecting a concentration difference on a fibre level. Proteins that promote particular functions such as growth or slow phenotype are not necessarily higher in muscles with that particular trait (e.g. higher in larger fibres or slow muscle). Interindividual differences in fibre composition might explain variable responses to training and insulin. PMID- 15362494 TI - Tumour-stroma interactions in carcinogenesis: basic aspects and perspectives. AB - In contrast to the conventional notion regarding tumour development as a cell autonomous process in which the major participants were the cancer cells, increasing evidence attributes important role in the stromal components, namely fibroblasts, and view the tumour as a heterogenous mixture of different cell types. These different types of cells, being cancer cells, fibroblasts, endothelial cells, and others, interact reciprocally and play an almost equally important role in the manifestation of certain aspects of the malignant phenotype. The elucidation of the mechanistic base of such interactions, besides the contribution to understand fundamental aspects of tumour cell biology, promises important applications in diagnosis, prognosis and therapy of the disease. PMID- 15362495 TI - Effects of alpha-lipoic acid supplementation on maternal diabetes-induced growth retardation and congenital anomalies in rat fetuses. AB - The mechanism of diabetic embryopathy is not known. Excessive reactive oxygen species (ROS) produced in diabetes may be causally related to foetal anomalies. The objective of this study was to determine whether supplementation with the antioxidant lipoic acid (LA) could prevent maternal diabetes-related foetal malformations and intrauterine growth retardation (IUGR) in rats. Pregnant rats were non-treated (Group I) or made diabetic on gestation day (GD) 2 by injecting streptozotocin (Group II). Group III was injected with 20 mg kg(-1) of LA daily starting on GD 6 and continued through GD 19. Group IV was administered only Tris buffer on the corresponding days. Group V was a set of STZ-treated animals, which were supplemented with a daily dose of 20 mg kg(-1) of LA from GD 6 through GD 19. All fetuses were collected on GD 20. Lipoic acid did not affect the blood sugar levels of diabetic animals significantly but improved their body weight gain and reduced food and water consumption. Diabetic group had a high incidence of embryonic resorption, IUGR, craniofacial malformations, supernumerary ribs and skeletal hypoplasia. Lipoic acid significantly reduced these abnormalities. These data support the hypothesis that ROS are causally related to fetal maldevelopment and IUGR associated with maternal diabetes in the rat. They also highlight the possible role of antioxidants in the normal processes of embryo survival, growth and development. PMID- 15362496 TI - Effects of diabetes mellitus on salivary secretion and its composition in the human. AB - This study investigated the effects of diabetes mellitus (types I and II) on human salivary gland function compared to healthy age-matched controls. The results have shown that both type I and type II diabetic patients secrete significantly (p < 0.05) less resting and stimulated saliva compared to healthy age-matched controls (AMC). It was also found that the diabetic patients have an increased resting and stimulated salivary protein concentration compared to healthy participants. However, the secretory capacity (stimulated minus resting values) was markedly reduced compared to controls. The level of calcium (Ca2+) in the saliva of diabetic patients was significantly (p < 0.05) elevated compared to the AMC. In contrast, the levels of magnesium (Mg2+), zinc (Zn2+) and potassium (K+) in the saliva of diabetic patients were significantly (p < 0.05) reduced compared to the values obtained in AMC. These results indicate that diabetes mellitus can lead to marked dysfunction of the secretory capacity of the salivary glands. In these patients a modified fluid, organic and inorganic salivary secretion may be responsible for the increased susceptibility to oral infections and impaired wound healing described by others in the literature. PMID- 15362497 TI - Biochemical effects of Citrullus colocynthis in normal and diabetic rats. AB - Diabetes mellitus is one of the most common endocrine diseases. In UAE many traditional plants such as the Citrullus colocynthis (Handal) are used as antidiabetic remedies. The aim of this study was to examine the effect of the aqueous extract of the seed of C. colocynthis on the biochemical parameters of normal and streptozotocin (STZ)-induced diabetic rats. Diabetes mellitus was induced by a single intraperitoneal (60 mg/kg body wt1) injection of STZ. Normal and diabetic rats were fed with the plant extract daily by oral intubation for 2 weeks. Blood sample were collected at the beginning and end of the experiment for the measurement of biochemical parameters. The plasma level of alanine aminotranferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), lactic dehydrogenase (LDH) increased significantly after the onset of diabetes. Oral administration of the plant extract reduced the plasma level of AST and LDH significantly. However, the plant extract failed to reduce the increased blood level of GGT and ALP in diabetic rats. Blood urea nitrogen (BUN) increased significantly after the onset of diabetes. No significant difference was observed in the blood creatinine, K+, Na+, Ca2+ and P levels of normal and diabetic rats. The plant extract did not have any effect on BUN level, however, it caused an increase in the level of K+, Na+ in diabetic rats. In conclusion, oral administration of the aqueous extract of the C. colocynthis can ameliorate some of the toxic effects of streptozotocin. PMID- 15362498 TI - Role of calpains in diabetes mellitus-induced cataractogenesis: a mini review. AB - Premature visual impairment due to lens opacification is a debilitating characteristic of untreated diabetes. Lens opacification is primarily due to the insolubilization of crystallins, proteins essential for lens optical properties, and recent studies have suggested that a major cause of this insolubilization may be the unregulated proteolysis of crystallins by calpains. These are intracellular cysteine proteases whose activation requires the presence of calcium (Ca2+) and elevated levels of lens Ca2+ is a condition associated with both diabetic cataractogenesis and other forms of the disorder. A number of calpains have been identified in the lens, including calpain 2, calpain 10 and two isozymes of calpain 3: Lp82 and Lp85. The use of animal hereditary cataract models have suggested that calpain 2 and/or Lp82 may be the major calpains involved in murine cataractogenesis with contributions from calpain 10 and Lp85. However, calpain 2 appears to be the major calpain involved in murine diabetic cataractogenesis and the strongest candidate of the calpains for a role in human types of cataractogenesis. Here, we present an overview of recent evidence on which these observations are based with an emphasis on the ability of calpains to proteolyse lens crystallins and calpain structural features, which appear to be involved in the Ca2+-mediated activation of these enzymes. PMID- 15362499 TI - Role of calpains in diabetes mellitus: a mini review. AB - Type 2 diabetes mellitus (T2DM) is characterized by defects in haepatic glucose production, insulin action and insulin secretion, which can also lead to a variety of secondary disorders. The disease can lead to death without treatment and it has been predicted that T2DM will affect 215 million people world-wide by 2010. T2DM is a multifactorial condition whose precise genetic causes and biochemical defects have not been fully elucidated but at both levels, calpains appear to play a role. Positional cloning studies mapped T2DM susceptibility to CAPN10, the gene encoding the intracellular cysteine protease, calpain 10. Further studies have shown a number of non-coding polymorphisms in CAPN10 to be functionally associated with T2DM whilst the identification of coding polymorphisms, suggested that mutant calpain 10 proteins may also contribute to the disease. The presence of both calpain 10 and its mRNA have been demonstrated in tissues from several mammalian species whilst calpain 10 appears to be associated with pathways involved in glucose metabolism, insulin secretion and insulin action. It appears that other calpains may also participate in these pathways and here we present an overview of recent studies on calpains and their putative role in T2DM. PMID- 15362500 TI - The in vitro retardation of porcine cataractogenesis by the calpain inhibitor, SJA6017. AB - Calpain inhibitors show the potential to serve as non-surgical alternatives in treating diabetic cataract and other types of these disorders. Here, we have tested the recently developed calpain inhibitor, SJA6017, for its ability to inhibit cataractogenesis in porcine lenses. These lenses were incubated in increasing levels of extralenticular calcium (Ca2+; 5-30 mM). Atomic absorption spectroscopy was used to determine total internal lens Ca2+ and a correlation between porcine lens Ca2+ uptake and levels of lens opacification were found with a total internal lens Ca2+ level of 5.8 microM Ca2+ g(-1) wet lens weight corresponding to the onset of catarctogenesis. A total internal lens Ca2+ level of 8.0 microM Ca2+ g(-1) wet lens weight corresponded to cataract occupying approximately 70% of the lens cell volume. This degree of cataract was reduced by approximately 40%, when SJA6017 (final concentration 0.8 microM) was included in the extralenticular medium, suggesting that the Ca2+-mediated activation of calpains may be involved in the observed opacification. Supporting this suggestion atomic absorption spectroscopy showed that the effect of SJA6017 (final concentration 0.8 microM) on lens opacification was not due to the compound restricting porcine lens Ca2+ uptake. The results indicate that calpain induced cataractogenesis is dependent on extracellular Ca2+ and the calpain inhibitor SJA6017 (0.8 microM) had no significant effect on Ca2+ uptake by lens. Its inhibitory effect on lens opacification may be due to a direct action on the activity of calpain. PMID- 15362501 TI - Streptozotocin-induced type 1 diabetes mellitus alters the morphology, secretory function and acyl lipid contents in the isolated rat parotid salivary gland. AB - Diabetes mellitus (DM) is associated with numerous conditions including hypo secretion of digestive enzymes. This study investigated the morphology, secretory function (alpha-amylase release) and acyl lipid contents in the isolated parotid gland of STZ-induced diabetic and age-matched control rats in order to provide insights into diabetes-induced salivary insufficiency. The techniques employed included light microscopy, colourimetric and gas chromatography (GC) analysis, respectively. Diabetes mellitus was induced in adult male Wistar rats by a single intraperitoneal (IP) injection of streptozotocin (STZ) (60 mg per kg body weight). Control animals were injected with a similar volume of citrate buffer. The animals were tested for DM 4 days after STZ injection and 2 months later when they were humanely killed for the experiment. The morphological results showed diabetic parotid glands to be extensively infiltrated with lipid droplets of various magnitudes, whereas glands from control animals display normal structure with the absence of lipid droplets. The analysis of parotid secretory function revealed a significant (p < 0.05) dose-dependent decrease in alpha-amylase release in response to noradrenaline (NA) in STZ-treated glands when compared to age-match control parotid glands. Furthermore, the levels of acyl lipids (16:0, 16:1, 18:0 and 18:1) in diabetic parotid glands was significantly (p < 0.01) reduced compared to control glands, along with a reduced ratio of 16:1/16:0. The results indicate DM can elicit changes in the morphology, secretory function and acyl fatty acid quantity in the isolated rat parotid gland. PMID- 15362502 TI - The English national risk-reduction programme for preservation of sight in diabetes. AB - This paper discusses the new national guidelines for a systematic screening programme to detect sight-threatening diabetic retinopathy in the population of people with diabetes in England. A review of the literature examines the evidence base to support screening interventions and effective management and treatments in diabetic retinopathy. The current evidence supports the establishment of a digital retinal photography system using pupil dilation. A Policy Advisory Group has been formulated by the National Screening Committee to guide the meeting of this target in England. A conclusion is made that with increased effort and organisation, health care professionals can ensure that the screening programme is successfully implemented and rates of visual impairment and blindness caused by diabetic retinopathy can be reduced significantly. PMID- 15362503 TI - Molecular and cellular basis of the aetiology and management of diabetic cardiomyopathy: a short review. AB - Diabetes mellitus is one of the most common chronic diseases affecting millions of people worldwide. Cardiovascular complication including myocardial infarction is one of the major causes of death in diabetic patients. Diabetes mellitus induces abnormal pathological findings including cell hypertrophy, neuropathy, interstitial fibrosis, myocytolysis and apoptosis and lipid deposits in the heart. In addition, the cytoplasmic organelles of cardiomyocytes including the plasma membrane, mitochondrion and sarcoplasmic reticulum are also impaired in both type I and type II diabetes. Hyperglycaemia is a major aetiological factor in the development of diabetic cardiomyopathy in patients suffering from diabetes. Hyperglycaemia promotes the production of reactive oxygen (ROS) and nitrogen species (RNS). The release of ROS and RNS induces oxidative stress leading to abnormal gene expression, faulty signal transduction and apoptosis of cardiomyocytes. Hyperglycaemia also induces apoptosis by p53 and the activation of the cytochrome c-activated caspase-3 pathway. Stimulation of connective tissue growth factor and the formation of advanced glycation end products in extracellular matrix proteins induces collagen cross-linking and contribute to the fibrosis observed in the interstitium of the heart of diabetic subjects. In terms of signal transduction, defects in intracellular Ca2+ signalling due to alteration of expression and function of proteins that regulate intracellular Ca2+ also occur in diabetes. All of these abnormalities result in gross dysfunction of the heart. Beta-adrenoreceptor antagonists, ACE inhibitors, endothelin-receptor antagonist (Bonestan), adrenomedullin, hormones (insulin, IGF 1) and antioxidants (magniferin, metallothionein, vitamins C and E) reduce interstitial fibrosis and improve cardiac function in diabetic cardiomyopathy. PMID- 15362504 TI - Defective sarcolemmal phospholipase C signaling in diabetic cardiomyopathy. AB - Phospholipase C (PLC) activity is known to influence cardiac function. This study was undertaken to examine the status of PLC beta3 in the cardiac cell plasma membrane (sarcolemma, SL) in an experimental model of chronic diabetes. SL membrane was isolated from diabetic rat hearts at 8 weeks after a single i.v. injection of streptozotocin (65 mg/kg body weight). The total SL PLC was decreased in diabetes and was associated with a decrease in SL PLC beta3 activity, which immunofluorescence in frozen diabetic left ventricular tissue sections revealed to be due to a decrease in PLC beta3 protein abundance. In contrast, the SL abundance of Gqalpha was significantly increased during diabetes. These changes were associated with a loss of contractile function (+/- dP/dt). A 2-week insulin treatment of 6-week diabetic animals partially normalized all of these parameters. These findings suggest a defect in PLC beta3 mediated signaling processes may contribute to the cardiac dysfunction seen during diabetes. PMID- 15362505 TI - Physiological effects and biochemical properties of a serum protein that produces positive inotropic and chronotropic effects on isolated guinea pig atria. AB - Isolated left and right guinea pig atria were used as a bioassay for the detection of an endogenous cardioactive substance in bovine serum. Serum, buffer exchanged to Krebs-Henseleit solution, produced positive inotropic and chronotropic effects on the isolated guinea pig atria. The cardiotonic effects were unaffected by the combined presence of propranolol and methysergide (both 10(-6)M) and were also dissimilar in time course from other known cardiotons such as catecholamines and cardiac glycosides. Following ultrafiltration (using XM100A Amicon membranes), activity was found solely in the retentate fractions and was therefore probably due to a large molecular weight (> 100 kDa) substance or a small molecule bound to a large protein. The cardioactive factor (CF) in the whole serum was heat labile, sensitive to acidification, exposure to potassium bromide and equilibration to physiological buffers of a low ionic strength. Isolation by conventional protein purification techniques was unsuccessful due to the labile nature of the active molecule(s) when exposed to non-physiological experimental conditions. Physical and biochemical properties of the CF which may help avoid inactivation are discussed for future experiments aimed at elucidating the nature and identity of the cardiotonic principle. PMID- 15362506 TI - Effects of halothane, isoflurane, sevoflurane and desflurane on contraction of ventricular myocytes from streptozotocin-induced diabetic rats. AB - Various clinically used volatile general anaesthetics (e.g. sevoflurane, halothane, isoflurane and desflurane) have been shown to have significant negative inotropic effects on normal ventricular muscle. However, little is known about their effects in ventricular tissue from diabetic animals. Streptozotocin (STZ)-induced diabetes is known to induce changes in the amplitude and time course of shortening and one report suggests that the inotropic effects of anaesthetics are ameliorated in papillary muscles from diabetic animals. The aim of these studies was to investigate this further in electrically stimulated (1 Hz) ventricular myocytes. Cells were superfused with either normal Tyrode (NT) solution or NT containing anaesthetic (1 mM) for a period of 2 min (at 30-32 degrees C). Myocytes from STZ rats were shown to have a significantly longer time to peak shortening (p > 0.001, n = 50) and the amplitude of shortening tended to be greater but this was not significant (p = 0.13, n = 50). Halothane, isoflurane, desflurane and sevoflurane significantly (p < 0.05) reduced the magnitude of shortening of control cells by 72.5 +/- 3.2%, 46.5 +/- 9.7%, 28.9 +/ 4.3% and 22.8 +/- 5.6%, respectively (n > 11 per group) but their steady-state negative inotropic effect was found to be no different in cells from STZ-treated rats (73.0 +/- 4.8%, 40.7 +/- 4.7%, 25.0 +/- 5.2% and 19.8 +/- 5.2%, respectively, n > 10 per group). Therefore, we conclude that the inotropic effects of volatile anaesthetics were not altered by STZ treatment. PMID- 15362507 TI - Effects of pressure overload-induced hypertrophy on TTX-sensitive inward currents in guinea pig left ventricle. AB - We investigated the effects of pressure overload hypertrophy on inward sodium (I Na) and calcium currents (I Ca) in single left ventricular myocytes to determine whether changes in these current systems could account for the observed prolongation of the action potential. Hypertrophy was induced by pressure overload caused by banding of the abdominal aorta. Whole-cell patch clamp experiments were used to measure tetrodotoxin (TTX)-sensitive inward currents. The main findings were that I Ca density was unchanged whereas I Na density after stepping from -80 to -30 mV was decreased by 30% (-9.0 +/- 1.16 pA pF(-1) in control and -6.31 +/- 0.67 pA pF(-1) in hypertrophy, p < 0.05, n = 6). Steady state activation/inactivation variables of I Na, determined by using double-pulse protocols, were similar in control and hypertrophied myocytes, whereas the time course of fast inactivation of I Na was slowed (p < 0.05) in hypertrophied myocytes. In addition, action potential clamp experiments were carried out in the absence and presence of TTX under conditions where only Ca2+ was likely to enter the cell via TTX-sensitive channels. We show for the first time that a TTX sensitive inward current was present during the plateau phase of the action potential in hypertrophied but not control myocytes. The observed decrease in I Na density is likely to abbreviate rather than prolong the action potential. Delayed fast inactivation of Na+ channels was not sustained throughout the voltage pulse and may therefore merely counteract the effect of decreased I Na density so that net Na+ influx remains unaltered. Changes in the fast I Na do not therefore appear to contribute to lengthening of the action potential in this model of hypertrophy. However, the presence of a TTX-sensitive current during the plateau could potentially contribute to the prolongation of the action potential in hypertrophied cardiac muscle. PMID- 15362508 TI - Effects of acidosis on ventricular myocyte shortening and intracellular Ca2+ in streptozotocin-induced diabetic rats. AB - We have investigated the effects of acute acidosis on ventricular myocyte shortening and intracellular Ca2+ in streptozotocin (STZ)-induced diabetic rat. Shortening and intracellular Ca2+ were measured in electrically stimulated myocytes superfused with either normal Tyrode solution pH adjusted to either 7.4 (control solution) or 6.4 (acid solution). Experiments were performed at 35-36 degrees C. At 8-12 weeks after treatment, the rats that received STZ had lower body and heart weights compared to controls, and blood glucose was characteristically increased. Contractile defects in myocytes from diabetic rat were characterized by prolonged time to peak shortening. Superfusion of myocytes from control and diabetic rats with acid solution caused a significant reduction in the amplitude of shortening; however, the magnitude of the response was not altered by STZ treatment. Acid solution also caused significant and quantitatively similar reductions in the amplitude of Ca2+ transients in myocytes from control and diabetic rats. Effects of acute acidosis on amplitude of myocyte contraction and Ca2+ transient were not significantly altered by STZ treatment. Altered myofilament sensitivity to Ca2+ and altered mechanisms of sarcoplasmic reticulum Ca2+ transport might partly underlie the acidosis-evoked reduction in amplitude of shortening in myocytes from control and STZ-induced diabetic rat. PMID- 15362509 TI - Voltage-dependence of contraction in streptozotocin-induced diabetic myocytes. AB - Cardiac contractile dysfunction is frequently reported in human patients and experimental animals with type-1 diabetes mellitus. The aim of this study was to investigate the voltage-dependence of contraction in ventricular myocytes from the streptozotocin (STZ)-induced diabetic rat. STZ-induced diabetes was characterised by hyperglycaemia and hypoinsulinaemia. Other characteristics included reduced body and heart weight and raised blood osmolarity. Isolated ventricular myocytes were patched in whole cell, voltage-clamp mode after correcting for membrane capacitance and series resistance. From a holding membrane potential of -40 mV, test pulses were applied at potentials between -30 and +50 mV in 10 mV increments. L-type Ca2+ current (I Ca,L) density and contraction were measured simultaneously using a video-edge detection system. Membrane capacitance was not significantly altered between control and STZ induced diabetic myocytes. The I Ca,L density was significantly (p < 0.05) reduced throughout voltage ranges (-10 mV to +10 mV) in myocytes from STZ-treated rats compared to age-matched controls. Moreover, the amplitude of contraction was significantly reduced (p < 0.05) in myocytes from STZ-treated rats at all test potentials between -20 mV and +30 mV. However, in electrically field-stimulated (1 Hz) myocytes, the amplitude of contraction was not altered by STZ-treatment. It is suggested that in field-stimulated myocytes taken from STZ-induced diabetic hearts, prolonged action potential duration may promote increased Ca2+ influx via the sodium-calcium exchanger (NCX), which may compensate for a reduction in Ca2+ trigger through L-type-Ca2+-channels and lead to normalised contraction. PMID- 15362510 TI - Increased inhibition of SERCA2 by phospholamban in the type I diabetic heart. AB - The sarcoplasmic reticulum (SR) plays a critical role in mediating cardiac contractility and its function is abnormal in the diabetic heart. However, the mechanisms underlying SR dysfunction in the diabetic heart are not clear. Because protein phosphorylation regulates SR function, this study examined the phosphorylation state of phospholamban, a key SR protein that regulates SR calcium (Ca2+) uptake in the heart. Diabetes was induced in male Sprague-Dawley rats by an injection of streptozotocin (STZ; 65 mg kg(-1) i.v.), and the animals were humanely killed after 6 weeks and cardiac SR function was examined. Depressed cardiac performance was associated with reduced SR Ca2+-uptake activity in diabetic animals. The reduction in SR Ca2+-uptake was consistent with a significant decrease in the level of SR Ca2+-pump ATPase (SERCA2a) protein. The level of phospholamban (PLB) protein was also decreased, however, the ratio of PLB to SERCA2a was increased in the diabetic heart. Depressed SR Ca2+-uptake was also due to a reduction in the phosphorylation of PLB by the Ca2+-calmodulin dependent protein kinase (CaMK) and cAMP-dependent protein kinase (PKA). Although the activities of the SR-associated Ca2+-calmodulin-dependent protein kinase (CaMK), cAMP-dependent protein kinase (PKA) were increased in the diabetic heart, depressed phosphorylation of PLB could partly be attributed to an increase in the SR-associated protein phosphatase activities. These results suggest that there is increased inhibition of SERCA2a by PLB and this appears to be a major defect underlying SR dysfunction in the diabetic heart. PMID- 15362511 TI - Halothane alters contractility and Ca2+ transport in ventricular myocytes from streptozotocin-induced diabetic rats. AB - General anaesthetics have previously been shown to have profound effects on myocardial function. Moreover, many patients suffering from diabetes mellitus are anaesthetised during surgery. This study investigated compromised functioning of cardiac myocytes from streptozotocin (STZ)-induced diabetic rats and the additive effects of halothane on these dysfunctions. Ventricular myocytes were isolated from 8 to 12 weeks STZ-treated rats. Contraction and intracellular free calcium concentration ([Ca2+]i) were measured in electrically field-stimulated (1 Hz) fura-2-AM-loaded cells using a video-edge detection system and a fluorescence photometry system, respectively. L-type Ca2+ current was measured in whole cell, voltage-clamp mode. Halothane significantly (p < 0.01) depressed the amplitude and the time course of the Ca2+ transients in a similar manner in myocytes from control and STZ-treated rats. However, the effect of halothane on the amplitude of shortening and L-type Ca2+ current was more pronounced in myocytes from STZ treated animals compared to age-matched controls. Myofilament sensitivity to Ca2+ was significantly (p < 0.01) increased in myocytes from STZ-treated rats compared to control. However, in the presence of halothane the myofilament sensitivity to Ca2+ was significantly (p < 0.05) reduced to a greater extent in myocytes from STZ-treated rats compared to controls. In conclusion, these results show that contractility, Ca2+ transport and myofilament sensitivity were all altered in myocytes from STZ-treated rats and these processes were further altered in the presence of halothane suggesting that hearts from STZ-induced diabetic rats are sensitive to halothane. PMID- 15362512 TI - The prevalence, type and severity of cardiovascular disease in diabetic and non diabetic patients: a matched-paired retrospective analysis using coronary angiography as the diagnostic tool. AB - People with diabetes mellitus have a 2-8-fold excess in cardiovascular mortality than people without diabetes. This study compared angiographically determined cardiovascular disease in 79 patients with diabetes mellitus and an equal number of matched controls without diabetes under the age of 55 years. Seventy-nine diabetic patients coming to coronary angiography during a 12-month period were reviewed retrospectively along with 79 control patients matched for age (+/- 3 years), sex, ethnic origin and risk factors (hyperlipidemia, body mass index and smoking history). The angiographic features of a consecutive series of 62 European and 17 Asian patients and their matched-paired controls were assessed. In all study subjects had undergone elective coronary angiography and ventriculography. Angiographic findings were graded to describe severity and extent of coronary atherosclerosis. Left ventricular systolic function was assessed by ejection fraction. The diabetic group had a significantly higher arterial systolic pressure than the non-diabetic group (p < 0.008) and they were clinically obese with a body mass index of >30. Detailed analysis of the angiograms showed that prevalence and severity of coronary artery disease in diabetic patients was greater. The mean 'severity score' was 11.66 for the diabetic group against 8.49 for the non-diabetic group (p < 0.037). Multivessel disease was more common in diabetic patients than in the controls, with three vessel disease being the most common. Furthermore, 38 of 79 diabetic patients had three-vessel disease compared to 29 of 79 controls. Diabetic patients were also more likely to have more segments diseased in one vessel. Systolic function was reduced in the diabetic group, with a significantly lower (p < 0.05) mean ejection fraction. The present study supports the evidence that diabetic patients have more extensive coronary artery disease than non-diabetic patients and a poorer prognosis, and that the coronary arteries of the Asian patients were affected more adversely than those of the European group irrespective of the diabetic state. PMID- 15362513 TI - Renin-angiotensin blockade attenuates cardiac myofibrillar remodelling in chronic diabetes. AB - Previous studies have shown that the renin-angiotensin system (RAS) is activated in diabetes and this may contribute to the subcellular remodelling and heart dysfunction in this disease. Therefore, we examined the effects of RAS blockade by enalapril, an angiotensin-converting enzyme inhibitor, and losartan, an angiotensin receptor AT1 antagonist, on cardiac function, myofibrillar and myosin ATPase activity as well as myosin heavy chain (MHC) isozyme expression in diabetic hearts. Diabetes was induced in rats by a single injection of streptozotocin (65 mg/kg; i.v.) and these animals were treated with and without enalapril (10 mg/kg/day; oral) or losartan (20 mg/kg/day; oral) for 8 weeks. Enalapril or losartan prevented the depressions in left ventricular rate of pressure development, rate of pressure decay and ventricular weight seen in diabetic animals. Both drugs also attenuated the decrease in myofibrillar Ca2+ ATPase, Mg2+-ATPase and myosin ATPase activity seen in diabetic rats. The diabetes-induced increase in beta-MHC content and gene expression as well as the decrease in alpha-MHC content and mRNA levels were also prevented by enalapril and losartan. These results suggest the occurrence of myofibrillar remodelling in diabetic cardiomyopathy and provide evidence that the beneficial effects of RAS blockade in diabetes may be associated with attenuation of myofibrillar remodelling in the heart. PMID- 15362514 TI - [Male climacteric syndrome (andropause)]. AB - The concept of male climacteric syndrome or andropause was advanced 55 years ago based on the fact that some men aged over 50 would present similar clinical symptoms as menopausal women. Since then, many related concepts have been presented to describe this specific period of time in men's life, and controversies have never ceased on this concept and its implication. To represent this unique phenomenon in a very general manner, such terms as male climacteric syndrome (andropause), late-onset hypogonadism, and partial androgen deficiency in the aging male (PADAM) are extensively used in clinical practice at the present time. But strictly speaking, these terms differ significantly, each representing a specific physiological condition. Out of different objectives, researchers might use some of the concepts to design their experiments or summarize related data. But it is very important for them to clarify real intentions and particular characteristics of selected research objects, so as to choose and use a proper concept. PMID- 15362515 TI - [Immunomagnetic beads sorting and functional identification of human spermatogonial stem cells]. AB - OBJECTIVE: The very nature of spermatogonial stem cells (SSC) is still poorly understood. The objective of this study is to explore the specific markers of human SSC and search for the suitable method for their isolation and functional identification. METHODS: Adults testicular cell suspensions were sorted by immunomagnetic beads method using alpha6,beta1 integrin and Thy-1 markers. The light-scattering properties and DNA Ploidy of the resultant subpopulations were analyzed by flow cytometry. The efficacies of the sorting on human SSC were evaluated by germ cell transplantation. RESULTS: (1) The alpha6+ Thy-1+ c-kit- and beta1+ Thy-1+ c-kit- cells were relatively uniform subpopulations in size and morphology, which represented about 2%-3% and 0.5%-1% of the unsorted testis cells, respectively. The analysis of light-scattering properties showed that both of the subpopulations had low side light-scattering properties. The DNA Ploidy analysis showed significant changes of these two cell subpopulations in DNA Ploidy. The percentage of diploid cells in alpha6+ Thy-1+ c-kit- cell subpopulation significantly increased to 51.2% and synthesis phase and tetraploid cells disappeared. (2) The functional evaluation showed that the SSC in the alpha6+ Thy-1+ c-kit- cells were enriched 40 times and the SSC in the beta1+ Thy 1+ c-kit- cells 20 times that of the unsorted cells. CONCLUSION: The alpha6,beta1 integrin and Thy-1 may be used for the SSC isolation as positive markers. The immunomagnetic beads sorting using alpha6,beta1 integrin and Thy-1 markers can result in significant enrichment of human SSC. It will open up a wide prospect for the researches on the biology of human SSC and the treatment of male sterility. PMID- 15362516 TI - [Effect of injection of testosterone undecanoate and depot medroxyprogesterone acetate on the suppression of spermatogenesis]. AB - OBJECTIVE: To observe depot medroxyprogesterone acetate (DMPA) and testosterone undecanoate (TU) injected at 8-week intervals for the suppression of spermatogenesis in healthy Chinese men. METHODS: After screening, 30 healthy volunteers were enrolled and randomly assigned to 3 dosage-groups (n = 10/group): 1000 mg TU (Group A), 1000 mg TU plus 150 mg DMPA (Group B), 1000 mg TU plus 300 mg DMPA (Group C). All dosages were given as intramuscular injections at 8-week intervals. The study consisted of an 8-week control (baseline) period, a 24-week treatment period and a 24-week recovery period. RESULTS: Consistent azoospermia or severe oligozoospermia was achieved and maintained in all the volunteers during the treatment period, except 2 in the mere TU group who experienced a "rebound" in sperm concentrations. An 8-week regimen of TU plus DMPA at both tested combination dosages effectively suppressed spermatogenesis to azoospermia. All volunteers tolerated the injections; no serious adverse effects were reported. CONCLUSION: The lower combined dosage is recommended for further testing in an expanded clinical trial or contraceptive efficacy study. PMID- 15362517 TI - [Experimental research on the effect of arsenic trioxide on the growth of prostate cancer PC-3 cell lines]. AB - OBJECTIVE: To evaluate the effect of arsenic trioxide (As2O3) on the growth of human androgen-independent prostate cancer PC-3 cell lines. METHODS: The changes of the growth and shape of PC-3 cells cultured in As2O3 of different concentrations were observed under the optical microscope after using different concentrations of As2O3, and the growth inhibition curve of As2O3 was obtained by means of MTT. An analysis was made of the apoptosis of the PC-3 cells treated with different concentrations of As2O3 by flow cytometry. And the inhibiting mechanism of As2O3was investigated by means of Annexin-V-FIFC/PI double staining. RESULTS: Affected by As2O3, the PC-3 cells turned round and small and less transparent in cytoplasm, and some sheded and suspended in the culture medium. Forty-eight hours after treatment with different concentrations of As2O3 (7.8125 micromol/L, 15.625 micromol/L, 31.25 micromol/L, 62.5 micromol/L, 125 micromol/L, 250 micromol/L, 500 micromol/L), the growth inhibition rates of the PC-3 cells were (0.06+/-0.99)%, (15.01+/-1.12)%, (29.21+/-1.31)%, (34.32+/-1.14)%, (40.51+/ 1.81)%, (69.39+/-1.74)%, and (73.19+/- 2.41)% respectively; and 72 hours after treatment, the growth inhibition rates were (0.04+/-1.51)%, (16.19+/-1.04)%, (43.61+/-1.12)%, (56.66+/-1.23)%, (73.13+/-2.61)%, (5.22+/-1.74)% and (91.41+/ 2.81)%, respectively. Forty-eight hours after the administration of different concentrations of As2O3 (0.1 micromol/L, 1.0 micromol/L, 3.0 micromol/L, 5.0 micromol/L, 20.0 micromol/L, 50.0 micromol/L), the apoptosis rates were 0.87%, 5.33%, 8.94%, 9.66%, 12.56%, 45.59% and 69.09%, respectively; and 72 hours after the administration, the apoptosis rates were 0.13%, 13.49%, 4.96%, 11.10%, 20.72%, 92.92% and 93.61%, respectively. CONCLUSION: As2O3 could inhibit the growth of androgen-independent prostate cancer PC-3 cell lines. The inhibiting effect on PC-3 cell lines is correlated with the time of treatment and concentration of As2O3, and accelerates cell apoptosis. PMID- 15362518 TI - [Early diagnosis of prostate cancer using free/total prostate-specific antigen ratio with population-based screening data]. AB - OBJECTIVE: To evaluate the use of free/total prostate-specific antigen ratio (fPSA/tPSA ratio) in improving the early diagnosis of prostate cancer. METHODS: fPSA/tPSA ratio in the serum was analyzed prospectively in 187 men with tPSA ranging between 4.0 and 20.0 microg/L. All of them underwent ultrasound-guided sextant prostatic biopsies. Sensitivity, specificity, positive and negative predictive values were calculated by SPSS 10.0 software. RESULTS: Prostate cancer detection rates were 18.1% and 22.5% when tPSA was within the ranges of 4.0-10.0 g/L and 10.0-20.0 g/L respectively. fPSA/tPSA ratio was more significant than tPSA in all the men. When the cut-off value of fPSA/tPSA ratio was set at 0.25, 90.5% and 87.5% of cancers could be detected; and 26.7% and 11.3% of biopsies could be avoided within the tPSA ranges of 4.0-10.0 g/L and 10.0-20.0 g/L, respectively. CONCLUSION: The use of fPSA/tPSA ratio can improve prostate cancer detection and reduce unnecessary biopsies when tPSA is within the range of 4.0 10.0 microg/L and 10.0-20.0 microg/L. PMID- 15362519 TI - [How chronic prostatitis affects plasminogen activator system]. AB - OBJECTIVE: To elucidate how chronic prostatitis affects the expression and activity of the plasminogen activator (PA) system and relates to male infertility. METHODS: Twenty-three normal fertile males and 80 chronic prostatitis patients (40 fertile and 40 infertile) were included in this research. SDS polyacrylamide gel electrophoresis and fibrin overlay method were used to estimate the total PA, and tissue PA (tPA), urokinase type PA (uPA) in semen. RESULTS: Total PA, tPA and uPA highly expressed in normal males, but decreased in the semen of the chronic prostatitis patients of both the fertile and infertile groups. However, there was no significant difference in total PA between the fertile and infertile patients. CONCLUSION: Chronic prostatitis reduces the secretory function and PA synthesis and secretion of the prostate, but the decrease of PA alone does not cause infertility. PA may be one of the tools for estimating the function of the prostate. PMID- 15362520 TI - [Effect of topical application of a Rho-kinase inhibitor on the erectile response in rats]. AB - OBJECTIVE: To investigate the effect of Rho-kinase inhibitor applied topically on the penile erection and systemic circulation of rats. METHODS: Y-27632 was applied to the surface of the tunica albuginea or to the penile skin of rats, and the changes of CCP/MAP were observed continuously. RESULTS: Both methods of drug administration resulted in a marked increase in the erectile response both with and without stimulation of the innervation of the penile vasculature. Some doses of the drug were also found to reduce systemic blood pressure. CONCLUSION: Inhibitors of Rho-kinase may represent a new and promising method of treatment for erectile dysfunction. PMID- 15362521 TI - [Suprapubic prostatectomy by preset removable pouch suture and male sexual function]. AB - OBJECTIVE: To evaluate the effect of suprapubic prostatectomy by preset removable pouch suture on the sexual function of patients with benign prostate hyperplasia (BPH). METHODS: Retrospective study was made on 110 cases of BPH that underwent suprapubic prostatectomy by preset removable pouch suture between Jan. 2000 and Sep. 2003. The sexual function changes were evaluated before and after surgery by IIEF-5. RESULTS: Twelve cases of post-operative ED (7 mild, 5 severe) post operative were found in 58 cases with normal sexual function pre-operatively. Among the 30 cases of mild pre-operative ED, 4 severe after operation, 5 restored to normal erection (>21 scores according to IIEF-5), and 21 had no significant change, Of the 22 cases in with severe pre-operative ED, 7 had higher IIEF-5 scores (4 became mild, and 3 had >21 scores). The changes of IIEF-5 scores before and after operation were not statistically different, which could be observed in morning erection (68% pre-operatively and 75% post-operatively). Post-operative retrograde ejaculation was found in 16 case (19%). Eighty percent of the patients thought that the suprapubic prostatectomy by preset removable pouch suture had no effect on male sexual function. CONCLUSION: Suprapubic prostatectomy by preset removable pouch suture affects very little male's sexual function and is a safe method for the treatment of BPH patients. PMID- 15362522 TI - [Regulation of EGF-receptor expression by TGFalpha in human prostate androgen unresponsive cancer cells]. AB - OBJECTIVE: To elucidate the regulation of epidermal growth factor receptor (EGFR) expression by transforming growth factor (TGFalpha) and epidermal growth factor (EGF) in human prostate androgen-unresponsive cancer cells. METHODS: EGFR mRNA expression and its protein level were measured by means of RT-PCR and Western blot respectively in human prostate cancer androgen-unresponsive cell lines, ARCaP and PC3, all treated with exogenous TGFalpha. RESULTS: In the TGFalpha group, the levels of EGFR mRNA were 5.01 0.45 and 9.05 0.63 in PC3 and ARCaP respectively, significantly higher than in the control group (P < 0.05). The level of EGFR protein in PC3 treated with TGFalpha was 2.28 0.53, higher than in the control group (P < 0.05); however, the level of EGFR protein in ARCaP treated with TGFalpha was only 1.24 0.22, not different from the control (P > 0.05). CONCLUSION: TGFalpha/EGF-EGFR pathway serves as a key growth regulator in prostate cancer. TGFalpha, but not EGF, preferentially maintains an autocrine loop in human androgen-unresponsive prostate cancer. PMID- 15362523 TI - [Effect of chronic abacterial prostatitis on semen quality and efficacy of antibacterial agents]. AB - OBJECTIVE: To appraise the effect of chronic abacterial prostatitis (CAP) on semen quality and the efficacy of antibacterial agents. METHODS: The semen of 86 cases of CAP and 20 normal men were analysed together with the antisperm antibodies in their blood and semen, and the antisperm antibodies in the blood and spermatic fluid were determined. Then the 86 patients were randomized into two groups, one treated with Chinese traditional medicine, indomethacin and three kinds of antibacterial agents, the other with Chinese traditional medicine and indomethacin only. The treatment lasted 3 months, and a follow-up visit was paid every month. RESULTS: Sperm motility and normal modality of the CAP group were 59.8% and 52.8%, respectively, significantly lower than those of the normal group (P < 0.005). The positive rates of the antisperm antibodies in the blood and spermatic fluid were 46.5% and 50%, respectively, significantly higher than those of the normal group. The clinical efficacy was significantly better in the group treated with antibacterial agents than the one without (P < 0.05). CONCLUSION: CAP can affect sperm quality and decrease fertility. Antibacterial agents can obviously enhance sperm quality and the cure rate of CAP. PMID- 15362524 TI - [Applied study on small-dose growth hormone supplement for aging-related symptom complex]. AB - OBJECTIVE: To assess the response of aging-related symptom complex to small-dose supplement of growth hormone. METHODS: Thirty patients, aged 42 to 77, with aging related symptom complex and normal serum testosterone were studied. Each subject received growth hormone at the dosage of 0.04 U/kg, 3 times/week for six months. Before and after treatment, body weight, grip strength, abdomen circumference, IPSS, uroflow rate, prostate volume, serum testosterone, cholesterol, triglyceride, PSA, IGF-1, glucose and side effects were determined in every case. Self Rating Scale used by Bosphorus university was adopted. RESULTS: After the treatment, the rating scores were significantly improved, grip strength and serum level of IGF-1 increased, and abdomen circumference and serum cholesterol decreased. Whereas, IPSS, uroflow rate, prostate volume, serum testosterone, triglyceride, PSA, glucose and body weight were not significantly changed. CONCLUSION: Six months treatment with small-dose growth hormone can improve aging related symptom complex with little adverse effect. PMID- 15362525 TI - [Protection of melatonin against damage of sperm mitochondrial function induced by reactive oxygen species]. AB - OBJECTIVE: To study the damage of mitochondrial function of sperm induced by reactive oxygen species (ROS), and the protection of melatonin (MLT) against the damage. METHODS: Spermatozoa of normal physiological function selected from semen samples by Percoll gradient centrifugation technique were used as normal sperm models in the present study. Reactive oxygen species were generated by hypoxanthine xanthine oxidase system, and in the presence (or absence) of MLT (6 mmol/L), incubated with normal sperm models for 30 and 60 minutes. After incubation, the activity of succinate dehydrogenase (SDH) in mitochondria of spermatozoa was assessed by histochemical method, and spermatozoa were labeled with specific fluorescent probe of Rhodamine 123 to measure mitochondrial membrane potential (MMP) by flow cytometry. RESULTS: After normal spermatozoa were incubated with ROS, MMP of spermatozoa significantly decreased, and the activity of SDH almost decreased to zero. However, MLT had effect on reducing the damage of the mitochondrial function of sperm induced by ROS. CONCLUSION: ROS can damage the mitochondrial function of sperm by affecting MMP of spermatozoa and the activity of SDH. MLT can protect sperm mitochondria from the damage induced by ROS through its effective antioxidative potential. PMID- 15362526 TI - [Establishment of the animal model of induced high-cholesterol-atherosclerotic erectile dysfunction and the mechanisms of atherosclerotic erectile dysfunction]. AB - OBJECTIVE: To establish the animal model of atherosclerotic erectile dysfunction induced by high-cholesterol diet and explore the mechanisms of atherosclerotic erectile dysfunction. METHODS: Thirty male rabbits were divided into two groups randomly: normal diet (ND) group (n = 10) and high-cholesterol (HCH) group fed with 1.5% cholesterol diet (n = 20). Serum total cholesterol, plaque areas of the ascending aorta, the ratio of intima/media thickness and level of vascular cell adhesion molecule 1 (VCAM-1) mRNA expression of internal pudendal artery were determined after twelve weeks. RESULTS: After twelve weeks, all rabbits of the HCH group got hyperlipemia, 9 of which presented typical atherosclerosis (ATH). The rate of atherosclerosis induced by high-cholesterol diet was 52.9%. Serum total cholesterol levels of the ATH and HCH groups were higher than that of the ND group (P<0.01). The penile erection times and rate in the ATH and HCH groups decreased significantly, compared with the ND group (P<0.01). Both the ratio of intima/media thickness and the level of VCAM-1 mRNA expression of internal pudendal artery in the ATH group were higher than those in the HCH and ND groups (P<0.01). CONCLUSION: It is easy, prectuable and reliable to establish the animal model of atherosclerotic erectile dysfunction induced by high-cholesterol diet. And one of the main causes of atherosclerotic erectile dysfunction is hypercholesterolemia, which through VCAM-1 may induce the structural and functional modifications of the endothelium of the internal pudendal artery and make the corpus cavernosum ischemia. PMID- 15362527 TI - [Prostate cancer after prostatectomy for benign prostatic hyperplasia]. AB - OBJECTIVE: To study the diagnosis and treatment of prostate cancer after prostatectomy for benign prostatic hyperplasia (BPH). METHODS: Twelve cases of prostate cancer after prostatectomy for benign prostatic hyperplasia were reviewed and studied. The mean intervals between prostatectomy for BPH and the diagnosis of prostate cancer was 10 months to 14 years, 5.6 years on average. The main symptoms were dysuria, hematuria and pain. Serum prostate specific antigen (PSA) was elevated in 11 cases. Digital rectal examination (DRE) was abnormal in 8 cases. Three cases were in clinical stage B, 3 in stage C and 6 in stage D. Ten cases received combined androgen blockade therapy. Monotherapy with surgical castration was given to 2 cases. Three patients with urinary tract obstruction received additional treatment of TURP. RESULTS: During the 4 months to 8 years follow-up, 3 patients died, 6 remained stable and 2 deteriorated. CONCLUSION: Surgery for BPH could not prevent the development of prostate cancer. PSA and DRE were the main methods for the diagnosis of this prostate cancer. PMID- 15362528 TI - [Diagnosis of seminal duct system diseases by percutaneous vasography (a report of 70 cases)]. AB - OBJECTIVE: To evaluate the value of percutaneous vasography in the diagnosis of the disorders of the seminal duct system. METHODS: Percutaneous vasography was performed successfully in 63 male patients (24 cases of infertility, and 21 hemospermia, 16 chronic prostatitis, and prostate carcinoma) from November 1974 to December 2003. RESULTS: Vasography revealed abnormal ducts in 28 and disorders in 38 (60.3%), including 8 cases of sterility, 13 seminal vesiculitis, 16 chronic prostatitis 5 seminal vesical cyst, and 2 prostate carcinoma. CONCLUSION: Percutaneous vasography is an effective method to identify diseases of the seminal vesical and ductal system. PMID- 15362529 TI - [Analysis of the relative etiology of non-obstructive azoospermia]. AB - OBJECTIVE: To investigate the levels and changes of sexual hormone in serum and seminal plasma, and the epidermal growth factor (EGF), alpha-glucosidase, and fructose in the seminal plasma of non-obstructive azoospermic patients, and to assess the significance of the analysis of their chromosomes. METHODS: Thirty-six non-obstructive azoospermic males and 26 normal males were chosen. The results in reproductive endocrine and genetics obtained by radioimmunoassay, colorimetry, and chromosomes G and C banding assay were compared and analyzed to explore into the causes of infertility. RESULTS: The levels of FSH, LH, PRL and E2 in the serum of the non-obstructive azoospermic patients were higher than those of the control group, but the T level had no significant difference between the two groups. There were no obvious differences in the sexual hormones except in FSH of the seminal plasma. There were 2 cases of abnormal sex chromosomes, 2 cases of big Y chromosomes and 7 cases of small Y chromosomes in the non-obstructive azoospermic patients. No correlation was found between EGF and fructose in the seminal plasma. CONCLUSION: The level of sexual hormone in serum, alpha glucosidase in seminal plasma and abnormal chromosomes might be closely related to non-obstructive azoospermia. PMID- 15362530 TI - [Progress in Attractin gene]. AB - Attractin, the research of which has flourished in recent years, is an autosomal recessive gene. The gene is widely expressed, and involved in a number of physiological and pathological events. This paper reviews the research development of the Attractin gene, including the characterization, expression, mutation, function and molecular mechanism of this gene. Besides, further research on the gene is also suggested. PMID- 15362531 TI - [The role of the renin-angiotensin system in male reproduction]. AB - The circulating renin-angiotensin system (RAS) is well known for its role in the maintenance of blood pressure and electrolyte and fluid homeostasis. However, other local angiotensin-generating systems than the circulating RAS have been found in numerous tissues. The male reproductive system including the testis, epididymis, and prostate has several sites of intrinsic RAS activity. The local RAS in these tissues can be responsive to androgens, fat acid, drugs, and hypoxia. There has been evidence for the involvement of the RAS not only in male reproduction, but also in the development of prostate disease. Besides, the assessment of the local RAS activity may be helpful to the early diagnosis of tumor in the male reproductive system. PMID- 15362532 TI - [Antioxidant vitamin and male reproduction]. AB - Increased generation of ROS causes the lipid oxidation of the membrane of spermatozoa, but antioxidant vitamins play an important role in reproduction and help clear away ROS and protect the sperm membrane from lipid oxidation. This review focused on the effect of antioxidant vitamins on male reproduction and in the treatment of male infertility. PMID- 15362533 TI - [Effect of chuanxiongzine on the relaxation of isolated rabbit corpus cavernosum tissues]. AB - OBJECTIVE: To investigate the effects of chuanxiongzine on the relaxation of isolated rabbit corpus cavernosum tissues. METHODS: Observations were made on the relaxing effect of chuanxiongzine on phenylephrine (PE)-precontracted corpus cavernosum and cavernosal strips preincubated with L-NAME, and ODQ, using endothelium removal and experimental method of smooth muscle strips. The effect of chuanxiongzine on cAMP and cGMP levels in corpus cavernosum was measured by 125I radioimmunoassay. RESULTS: Chuanxiongzine had concentration-dependent relaxing effect on the cavernosal muscle contraction induced by PE (EC50 1.58 x 10(-4) mol/L). This relaxing effect was partially antagonized by ODQ and not blocked by L-NAME or endothelium removal. In PE-precontracted cavernosal strips, chuanxiongzine induced relaxation accompanied with an increase in cAMP and cGMP levels. CONCLUSION: Chuanxiongzine was effective in relaxing cavernosal muscle precontracted by PE in vitro. And its relaxing effect may be enhanced by increasing cGMP and cAMP levels in the corpus cavernosum. PMID- 15362534 TI - [Acute type A aortic dissection]. AB - BACKGROUND: Current surgical results for acute type A aortic dissection has been getting better, though the mortality rate still remains high, up to 10-15% in Japan. Our strategy consists of #1 make the most use of echography for better understanding of hemodynamics; #2 the extent of replacement depends on the site of initial entry; #3 deep hypothermic circulatory arrest and retrograde cerebral perfusion for brain protection. PATIENTS AND METHODS: Between January 2000 and December 2003, 45 consecutive patients were treated. Twelve of them had extended replacement of the arch, and 3 had simultaneous root replacement by means of a composite graft. RESULTS: In hospital mortality was 11% (5 patients). The cause of death was serious organ malperfusion, which had been developed preoperatively, in all patients but 1 who suffered from thoracic aortic obstruction due to newly developed type B aortic dissection. CONCLUSIONS: The determinant of surgical mortality in patients with acute type A aortic dissection is the presence of serious organ malperfusion. Rapid diagnosis and less invasive treatment for malperfusion is the key to improve the results. PMID- 15362535 TI - [Acute type B aortic dissection]. AB - Although most patients with acute type B aortic dissection are treated medically, emergency surgery is needed for patients with complications, such as rupture, uncontrollable pain, refractory hypertension, a huge pseudo-lumen or organ ischemia. Optimal surgery should be selected according to each condition. An emergent descending aortic replacement is applied mainly for ruptures or impending ruptures, while a surgical bypass, fenestration or thrombectomy is used for visceral organ or leg ischemia. In our data, 9 (23%) out of 40 patients with acute type B aortic dissection underwent emergent surgical treatment. In-hospital mortality was 11.1%. Because it is difficult to diagnose visceral ischemia, patients who might be affected with one must be treated promptly. Although catheter intervention is expected to improve the results of these high-risk patients, more investigation is needed to discover a new therapeutic strategy for acute type B aortic dissection. PMID- 15362536 TI - [Ruptured non-dissection thoracic aortic aneurysms]. AB - The number of patients with thoracic aortic aneurysm has been increasing in Japan. The annual report by the Japanese Association for Thoracic Surgery 2001 demonstrated that 4,133 patients underwent surgery for thoracic aortic aneurysm. Among them, ruptured cases consisted of 11% (460 patients) and its early mortality was 32%. At the emergency room, initial accurate diagnosis, using echocardiography and computed tomography (CT), is mandatory for the next step and catastrophic shock status should be corrected if possible, such as pericardiocentesis or transfusion. For patients with ruptured descending or thoracoabdominal aortic aneurysms, the endovascular stent-grafting should be first choice of treatment. Otherwise, patients should be sent for the operation room as quickly as possible for initialing cardiopulmonary bypass and for controlling bleeding. Special attention should be paid for not injuring the left lung after left thoracotomy. Mycotic false aneurysms, including the aorto esophageal fistula and prosthetic graft infection, remained as one of challenging entities in aortic emergency. The standard treatment for this fatal disease resection of the infected both aneurysm and esophagus, and has been reconstruction with an extra-anatomical bypass, however, recent advancement of technologies, including allograft and tissue engineering might enable to have in situ reconstruction of the aorta. PMID- 15362537 TI - [Traumatic rupture of the thoracic aorta]. AB - Nowadays, trauma victims are quickly transported from the scene to the therapeutic facility, therefore, even patients with traumatic aortic rupture (TAR) who were considered to be unlikely to survive several hours in the past are able to have more chance to undergo surgical treatment. In such patients who need emergency surgical repair of TAR regardless of the presence of multiple other organ injuries, massive contrast dye extravasation into the mediastinum is usually demonstrated on the contrast-enhanced computed tomography (CT) images. But, in most of the cases showing a pseudoaneurysm or dissection, delayed aortic repair is preferable when other organ injuries have therapeutic priority. The use of active adjunct means for distal support is definitely advantageous to prevent paraplegia. Heparin-bonded bypass by the use of low dose of heparin or intravenous argatroban is safe and avoids the risk of bleeding of other injured organs. PMID- 15362538 TI - [Acute infective endocarditis]. AB - Between January 1987 and June 2004, 65 patients with infective endocarditis underwent aortic and/or mitral surgery. Nine of these patients with active native valve endocarditis required emergency surgery. Indications for emergency operation were progressive congestive heart failure in 5 patients, systemic emboli in 2 patients and uncontrolled infection in 2 patients. The aortic valve was affected in 3 patients, the mitral valve in 4 patients and both the aortic and mitral valve in 2 patients. All of these patients were treated with radical excision of infected tissue and implantation of mechanical or stented bioprosthetic valves. There was no hospital death and no patients required reoperation for recurrent endocarditis. Although medical therapy for acute infective endocarditis is effective in most instances, some patients still require early surgical intervention. These data demonstrate excellent results in patients who underwent emergency surgery for acute infective endocarditis. PMID- 15362539 TI - [Thrombosed mechanical heart valve prostheses]. AB - Our clinical experiences of treatment for definite or probable thrombosed mechanical valve prostheses was presented and discussed concerning with surgical indication, several operative approaches to a target valve and detail surgical techniques. Preoperative diagnosis includes loss of metallic click sound, abnormal restricted disc movement and echocardiographic evidences of annular, occluder or housing thrombus. Valve re-replacement or thrombus removal was performed with re-do sternotomy or thoracotomy in urgent or elective cardiopulmonary bypass. Careful preparation of right atrium, superior and inferior vena cava and ascending aorta, and establishment of moderate hypothermic cardiopulmonary bypass were essential for re-replacement of the thrombosed valve. Among 15 patients, mitral re-replacement was performed in 10, aortic re replacement in 3, tricuspid re-replacement in 2 and thrombus removal in 1. Hospital mortality rate was 6.7%, and late recurrence of thrombosed valve was observed in a patient. Pathophysiology of thrombotic process and bioactivity on the prosthetic valves are yet unsolved. Further refinement of clinical strategy would be necessary in relation to the effect of antithrombotic and antiinflammatory treatment. PMID- 15362540 TI - [Left ventricular rupture following mitral valve replacement]. AB - Left ventricular rupture following mitral valve replacement is an unusual but highly lethal complication. Between 1982 and 2004, 254 patients underwent mitral valve replacement with coronary artery bypass grafting (CABG) [7 patients], aortic or tricuspid valve operation or both (84 patients), or without concomitant operation (163 patients). There were 11 hospital deaths (4.3%) and 4 left ventricular ruptures (1.6%). No concomitant operation had been performed in all rupture cases. Type I rupture occurred in 2 patients with complete resection of the mitral valve, and type III in the others with preservation of the posterior leaflet or basal chordae. Three of 4 were early and 1 was delayed rupture. The cause of rupture was obvious in 2 patients, over-excision of the posterior mitral annulus (type I) and impingement of the strut (type III). Intra-cardiac, extra cardiac or both combined surgical repair was attempted under the cardioplegic arrest for all patients, however, 2 of 4 patients died for postoperative low cardiac output syndrome (LOS) and intra-operative bleeding. Techniques of repair and prevention of rupture are discussed. PMID- 15362541 TI - [Acute mitral valve insufficiency caused by chordae rupture]. AB - The common causes of mitral valve chordae rupture are myxomatous valve disease, infective endocarditis. Acute mitral valve insufficiency from chordae rupture, severe clinical symptoms such as dyspnea and shock are observed. Diagnosis is made with clinical symptoms, chest X-ray and echocardiography. Echocardiography is very useful to evaluate the exact lesions. First, inotropic agents, dilators and intraaortic balloon pumping (IABP) are used to stabilize hemodynamic status. Once hemodynamic status becomes unstable in spite of such therapy, surgical correction of mitral regurgitation (MR) is considered. Mitral valve repair should be the first choice of surgical repair, because it preserves left ventricular function, avoids anti-coagulation therapy. Quadrangular resection and suture, annuloplasty and artificial chordae are reliable surgical techniques. If mitral valve repair is difficult, mitral valve replacement is another option. From 1994 to 2002, there were 12 patients who underwent emergency mitral valve surgery because of chordae rupture. There were 7 men and 5 women with mean age of 52.7+/ 14.3 years old. Nine of them received mitral valve repair. Two patients were infective endocarditis. Preoperative MR grade was 4.0+/-0.0. Postoperative MR grade was 0.2+/-0.2. Mean follow-up period was 3.8 years. There were no cardiac events and reoperations. Repair technique is quite reliable even in patients with multiple lesions, and the surgical results were almost same as single lesion patients. Repair technique should be the first choice to treat acute MR caused by chordae rupture. PMID- 15362542 TI - [Acute coronary syndrome]. AB - The first choice of interventional treatment for acute coronary syndrome is percutaneous coronary intervention (PCI), especially when the patient is in cardiogenic shock. In cases of acute myocardial infarction, emergency coronary artery bypass grafting (CABG) is indicated when left main trunk (LMT) lesion or severe triple-vessel disease is left after PCI combined with residual or recurrent ischemic angina and/or ST-T change in electrocardiogram (ECG). Similarly, in cases of unstable angina, emergency CABG is indicated when LMT or LMT equivalent (proximal left anterior descending branch and left circumflex branch) is the culprit lesion or when severe triple-vessel disease exists. Urgency of operation and existence of cardiogenic shock are major operative risk factors of CABG in patients with acute coronary syndrome. The surgical mortality of such cases is much higher than that of elective surgery for chronic angina. As employment of cardiopulmonary bypass (CPB) and achievement of cardioplegic arrest is one of the major causes of surgical mortality, off-pump CABG (OPCAB) using no CPB nor cardioplegic solution has recently become popular. OPCAB, however, is difficult to perform when the hemodynamic state is unstable. On-pump beating heart CABG is an alternative technique of choice because it not only stabilizes the hemodynamic state but also prevents myocardial ischemia/reperfusion injury. These beating heart CABG techniques will improve surgical outcome of acute coronary syndrome. PMID- 15362543 TI - [Iatrogenic myocardial infarction]. AB - In this paper, iatrogenic myocardial infarction resulting from percutaneous catheter intervention (PCI) and cardiac surgery is described. Among the patients who had experienced iatrogenic myocardial infarction in the past 4 years, 6 had undergone emergent coronary artery bypass grafting (CABG). An emergent operation for myocardial infarction was performed on 3 patients, for problems associated with PCI, and 1 patient had died due to cardiac failure. In the latter case, although the emergent CABG was performed after the failure of percutaneous coronary intervention in another hospital, no improvement in cardiac function was observed. The reason for this was that the myocardial damage became almost irreversible due to the delay in transferring the patient to our hospital. An additional CABG was performed in 2 cases, because they experienced a cardiogenic shock after cessation of the cardio-pulmonary bypass system. In these cases, cardiac function had improved immediately after an additional CABG. Some ST elevation in the electrocardiograms was observed in 8 cases in the intensive care unit after isolated CABG. One patient experienced a cardiogenic shock, and was transferred to the operating room immediately. The radial artery graft connected in sequence to circumflex coronary arteries was spastic, and an additional bypass was performed to the same coronary arteries using a saphenous vein graft. Although conservative medical therapy was administered in the remaining 7 cases, 1 case presented an occlusion of the left descending coronary artery, and an emergent CABG should have been performed. ST elevations in 2 cases were caused by coronary spasm, while in the remaining 4 cases, they were caused by pericardiotomy. Although a postoperative change in the electrocardiogram was considered important, the diagnosis was complicated. In myocardial infarction, cardiogenic shock develops easily, and an emergent CABG is needed in many cases. It is important to decide on an emergent operation promptly after an accurate diagnosis. PMID- 15362544 TI - [Ventricular septal perforation]. AB - As a mechanical complication of acute myocardial infarction, ventricular septal perforation (VSP) occurs mostly within 1 week after infarction. Although incidence of the disease is only 1-2% of transmural infarction, patients are into serious condition progressively once it happens. Even if patients were treated medically under intensive care protocol, they would almost always present congestive heart failure due to massive volume overload. Hence VSP should be treated surgically as soon as possible after its onset. We believe that infarct exclusion technique is standard procedure of surgical treatment, because it is relatively simple, makes less bleeding and gives minimal damage to right ventricle. Acute mortality has come down to 10-20% after introduction of this procedure. Cardiogenic shock before surgery, severe right heart failure and high age are important predictors of death. It is still difficult to save patients who are in severe cardiogenic shock with very low ventricular function. New treatments such as ventricular assist device, heart transplantation and regeneration therapy are waited for this high risk group of patients. PMID- 15362545 TI - [Papillary muscle rupture]. AB - Papillary muscle rupture (PMR) is a rare, but catastrophic mechanical complication of acute myocardial infarction (AMI). Patients with PMR present in acute pulmonary edema and commonly in cardiogenic shock. The absence of new heart murmur after AMI dose not exclude the diagnosis. To diagnose PMR accurately, transesophageal echocardiography is the most useful and essential, coronary angiography is also necessary for appropriate surgical treatment. Based on these results of diagnostic procedure, combined myocardial revascularization and mitral operation should be performed without delay. Although mitral valve repair has the effects of improving left ventricular function, repairing could be undertaken successfully in limited cases, such as ruptured muscle tissue is not friable. If repair is technically more challenging, mitral valve replacement should not be hesitated. Postoperative early and late survival may relate to surgical correction of coronary artery revascularization, especially in multivessel disease, concomitant coronary artery bypass grafting is benefit theoretically and should be performed. PMID- 15362546 TI - [Left ventricular free wall rupture]. AB - BACKGROUND: Left ventricular (LV) free wall rupture is an uncommon but catastrophic event after acute myocardial infarction and is associated with a high mortality. Some patients may be salvaged with emergent operation. Padro et al. described a sutureless technique for the treatment of postinfarction LV free wall rupture. We have adopted a similar sutureless technique using a fibrin glue oxycellulose fixation. METHODS: A review of 32 patients during a 20-year period who were referred to coronary care unit (CCU) with postinfarction LV free wall rupture was conducted. There were 17 men and 15 women with an average age of 68.8 years. Nine patients were treated who having one-stage operation of simultaneous 7 ventricular septal perforation and 2 coronary artery bypass grafting (CABG). Nine patients were sutured directly with a felt sandwich technique with or without cardiopulmonary bypass. Five patients were treated with pericardiectomy and pericardiocentesis using direct vision. Nine patients were sealed with sutureless method using fibrin glue-oxycellurose fixation without cardiopulmonary bypass. RESULTS: Eleven hospital deaths (34.4%, 11/32) were occurred in the all patints. However, 1 death (7.1%, 1/14) occurred in the pericardiectomy and in the sutureless sealed groups. CONCLUSIONS: A sutureless technique for an oozing type of postinfarction LV free wall rupture is effective. PMID- 15362547 TI - [Total anomalous pulmonary venous connection]. AB - The severity of symptoms in total anomalous pulmonary venous connection (TAPVC) depends primarily on the degree of pulmonary venous obstruction (PVO). With severe PVO, patients can present in extremis with severe cyanosis, respiratory distress, and acidosis within the first few hours of life. The diagnosis is usually established by two-dimensional echocardiography. Surgical repair of TAPVC has involved a number of techniques using various strategies of cardiopulmonary bypass and circulatory arrest, methods of cardioplegia, sites of venous cannulation, and cavitary exposures. Recent clinical investigations have uncovered some long-term negative effects of deep hypothermia and circulatory arrest strategies. Our current strategies for surgical management of TAPVC are 1) using moderate hypothermic cardiopulmonary bypass with bicaval venous cannulation combined with antegrade cardioplegia strategy and 2) performing the side-to-side anastomosis between the confluence of pulmonary veins and the left atrium by right side approach. The critical parts of this anastomosis are 1) making incisions of the confluence of the pulmonary veins and the left atrium to avoid the distortion of the anastomotic site and 2) performing the anastomosis without "purse string" the suture line. The prognosis of the patients with recurrent PVO is still developing; therefore great care should be taken at the initial procedure to avoid obstruction. PMID- 15362548 TI - [Pulmonary atresia with intact ventricular septum]. AB - It is important to employ a strategy for pulmonary atresia with intact ventricular septum (PAIVS) in collaboration with cardiology and cardiac surgery. We established our policy of treatment based on the right ventricular end diastolic volume. Sinusoidal communication should be carefully considered. Prognosis was poor in case of interruption of coronary arteries. Although catheter intervention is not fully established in initial treatment of PAIVS, it has a clinically significant roll in case of sufficient right ventricular end diastolic volume. PMID- 15362549 TI - [Transposition of the great arteries]. AB - This report describes changes in the treatment strategy for transposition of the great arteries (TGA) undergoing primary arterial switch operation early in the neonates in our institution. Decrease in pulmonary vascular resistance after birth results in deterioration of left ventricular (LV) performance as a systemic ventricle and right ventricular (RV) failure. Ductus arteriosus should be kept patent by alprostadil alfadex (PGE1) in order to prevent deterioration of LV performance. Contrarily, pulmonary high flow leads to the progression of RV failure. Balloon atrial septostomy (BAS) and PGE infusion may be helpful to manage severe hypoxemia, but an increase in aortic septal defect (ASD) flow may lead to deterioration of LV performance. To solve these dilemma primary arterial switch operation should be underwent as early as possible. PMID- 15362550 TI - [Hypoplastic left heart syndrome]. AB - Hypoplastic left heart syndrome (HLHS) is one of the last remaining problems in pediatric cardiac surgery, which necessitates a search for new solutions and continues to be a challenge for cardiologists and cardiac surgeons. The identification of HLHS in utero is extremely helpful for prompt, appropriate preoperative management. In addition, infants with prenatal diagnosis had an improvement operative mortality on Norwood operation compared to infants diagnosed postnatally. Outcome following Norwood procedure has improved significantly secondary to modifications in the surgical technique for protection of multiorgans, improved perioperative care, and improved anesthetic management. In particular right ventricular (RV)-pulmonary artery (PA) shunt using non-valved polytetrafluoroethylene (PTFE) graft was applied as a first stage palliation of HLHS to prevent hemodynamic instability associated with a classic Norwood procedure. In the modified Norwood (RV-PA shunt), higher diastolic blood pressures and lower Qp/Qs ratios that are associated with a more stable and efficient systemic circulation. In fact, patients who received a RV-PA shunt, rarely received ventilatory manipulations. PMID- 15362551 TI - [Coarctation of the aorta]. AB - The surgical outcome of neonatal and infantile coarctation of the aorta has been improved dramatically in recent years. Not only the improvement of perioperative management based on the better understanding of the pathophysiology but the improvement of the surgical procedure made the successful outcome. There are still, however, several controversial aspects of surgical management: How to select surgical technique? How to fix the distal or transverse aortic arch hypoplasia? How to approach the intracardiac defect? How to protect spinal cord? Postoperative recoarctation in adult age will be mostly concerned in recent years. Surgeons have to aware all these short- and long-term problems when fixing this particular cardiac lesion. Now I want to discuss this not-unusual and relatively complex heart disease in this article. PMID- 15362552 TI - [Arrhythmias]. AB - The success of the radiofrequency catheter ablation procedure for most types of supraventricular and ventricular tachycardia largely eliminated the role of surgical therapy of arrhythmias. However, there remains a subset of arrhythmia patients in whom urgent surgical treatments are required. In this review we mention recent developments of the urgent surgical treatment for arrhythmias. In cases with complete atrioventricular (AV) block and ventricular fibrillation which associated with sudden death, temporary cardiac pacing and cardiac defibrillation using direct current (DC) cardioversion must be immediately induced and followed by implantation of permanent cardiac pacemaker or implantable cardioverter defibrillator (ICD), if necessary. In addition to usual cardiac pacing therapy, novel pacing therapy has been developed recently for the patients with symptomatic heart failure. Cardiac resynchronization therapy (CRT) using biventricular pacing is an emerging therapy for improvement of cardiac function in patients with heart failure in association with intraventricular conduction delay. To prevent the sudden death in patients with heart failure, biventricular pacing combined with ICD are also implanted and its efficacy are reported. With the exception of the pacing therapy, curative surgical treatments are limited in drug-refractory atrial fibrillation and ventricular fibrillation/tachycardia after myocardial infarction requiring Dor's type operation. In any case surgical treatment must be performed promptly and suitably with lower invasive methods. PMID- 15362553 TI - [Pulmonary thromboembolism]. AB - Thromboembolism used to be a rare disease in Japan, however, more frequent occurrence and mortality was reported in recent periods. This is considered to relate with the rapid change of food intake, so called food westernization. The authors herein described the etiology and its countermeasures against acute and chronic pulmonary thromboembolism. Formation of thrombus relates with congestion, damage of venous vessels and change of blood component. Pulmonary thromboembolism during airflight, so called "economy syndrome" occurs due to the formation of thrombus at the large veins of lower extremities. Anti-coagulant treatment was applied first, and then surgical intervention was performed in acute phase. Pulmonary thromboembolectomy for chronic pulmonary thromboembolism is one of the most difficult techniques in thoracic surgery. Careful and cautious dissection between fibrous intima of the thrombus and media of the vein is needed. In conclusion, urgent and appropriate medical and surgical treatment is crucial to rescue the life of patients suffered from pulmonary thromboembolism. PMID- 15362554 TI - [Traumatic cardiac injury]. AB - Traumatic cardiac injuries are classified into sharp and blunt injuries and show various pathologic conditions. For patients' survival, each pathologic condition should be rapidly evaluated and appropriately treated. In many patients, direct suturing of the wound without using extracorporeal circulation is possible. However, in patients with intra-cardiac injury or severe free-wall injury, or patients in whom the operative field is difficult to obtain, repair under extracorporeal circulation is necessary. In our patients with sharp injury, the survival rate after surgery was 100%. However, surgical results in patients with blunt injury were markedly affected by the presence or absence of complication injuries, particularly head injury, and the survival rate after surgery was 72.7%. The survival rate may be improved by: (1) rapid and appropriate evaluation of cardiac injury in patients with multiple injuries, (2) prompt surgical treatment, and (3) comprehensive postoperative management and treatment for complication injuries. PMID- 15362555 TI - [Pneumothorax]. AB - Most of the patients with spontaneous pneumothorax are young male adults without significant pulmonary disease and the prognosis in these patients is usually very good. But in a few cases when tension pneumothorax develops, immediate diagnosis and chest tube drainage is required to avoid life-threatening emergency. Needless to say mechanical ventilation with tracheal intubation is contraindication even if the patient has severe respiratory distress or hypoxia. Re-expansion pulmonary edema is another severe complication. This occurs to a long-term (over 1 week) complete collapse lung when it is rapidly re-expanded with high negative pressure. To treat such patient it is necessary to decompress the lung slowly using lower negative pressure or water seal suction. When persistent bleeding of hemothorax is present, it is important to decide to perform open thoracotomy or video-assisted thoracoscopic surgery to avoid blood transfusion. The diagnosis of pneumothorax is simply done by chest X-ray film. When the pneumothorax is suspected, chest X-ray examination has to be done first. A scheduled operation to prevent recurrence of pneumothorax has to be performed under relative indication in consideration of both the medical factor and the social factor of the patient. PMID- 15362556 TI - [Chylothorax]. AB - Chylothorax is a rare complication after general thoracic surgery, but remains challenging for thoracic surgeons. Chylothorax can occur after any type of general thoracic procedures, including esophagectomy, pulmonary resection, and removal of mediastinal tumor. Unless being treated properly, chylothorax leads to high mortality. Therefore, prompt diagnosis and adequate therapy are crucial in the management of postoperative chylothorax. Chylothorax is usually discovered after a patient resumes taking fat-containing food. Milky fluid can be seen in the drainage system if a chest tube is still in place. When the diagnosis has been made, conservative treatment should be initiated at first, including cessation of oral intake and TPN. Evacuation of chylous fluid by closed chest tube drainage should also be done. A chest tube should be inserted in case the tube has already been taken out. If 10 to 14 days of conservative treatment fails, surgical treatment should be considered. Ligation or clipping of the thoracic duct itself or its tributaries is performed through open thoracotomy or video-assisted thoracoscopic (VATS) approach. The results of reoperation are usually satisfactory. PMID- 15362557 TI - [Traumatic lung injury]. AB - Pulmonary injuries include a wide variety of clinical conditions. Most patients with blunt chest trauma can be managed with conservative treatment. Only about 10 to 15% of patients with severe chest injuries require major thoracotomy. Management of pulmonary contusion, pulmonary laceration, pneumothorax or hemothorax by oxygen inhalation, respirator assist and chest drainage can usually result in complete recovery. However, pulmonary injuries sometimes lapse into fatal condition if they are improperly treated. Open thoracotomy is required in cases with persistent massive air leakage or massive bleeding with the use of chest drainage. It is crucial to evaluate the extent and severity of the injuries based on chest X-ray and computed tomography (CT) findings for the proper initial treatment in patients with pulmonary injuries. PMID- 15362558 TI - [Blunt tracheobronchial injury]. AB - Blunt tracheobronchial injury is rare but crucial injury. We discussed some problems about blunt tracheobronchial injury and presented our experience. To rescue patients with blunt tracheobronchial injury, surgical treatment within 24 hours from injury is recommended when general anesthesia is acceptable. Therefore, bronchoscopy should be performed immediately for patients who transferred to hospital due to blunt chest trauma. Outcome of surgery for tracheobronchial injury depends on co-existence of other organs and the degree of tracheobronchial injury itself. To improve the prognosis of patients with blunt trachobronchial injury, we need to know the accurate incidence and results of treatment in Japan. To know the fact, a large-scaled prospective registration of cases with tracheobronchial injury is necessary. At present, the Japanese association for the surgery of trauma is conducting such registration. We present a 29 year-old male who had blunt tracheobronchial injury and underwent reconstruction of carina in our hospital. Rupture of carina was recognized by bronchscopy and pneumomediastinum was shown in chest computed tomography (CT). Operation was successfully performed 22 hours later from injury and the patient discharged 28 days after from injury. PMID- 15362559 TI - [Bronchial bleeding]. AB - The acute and general management of patients with massive hemoptysis is described. Initial priorities are insuring adequate airway protection, ventilation, and cardiovascular function. Major priorities of ongoing hemoptysis to prevent gas exchange are intubation with a large bore endotracheal tube and protection of nonbleeding lung by selective intubation into nonbleeding main stem bronchus. Once the patient stabilized, bronchoscopic procedure to stop bleeding following by bronchial arterial embolization should be planned. While surgery remains only definitive therapy, it should not be used in the acute emergent setting unless it cannot be avoided. Bronchial artery and other feeding arteries in the chest wall should be shut off before surgery to prevent massive bleeding during operation. PMID- 15362560 TI - [Benign mediastinal tumors]. AB - Most of benign mediastinal tumors are asymptomatic; however, sometimes we encounter emergent cases. In this article we discuss with regard to the situation that benign mediastinal tumors with severe symptoms need to be treated emergently. Any benign mediastinal tumors can cause respiratory and cardiac failure as they grow large enough to compress surrounding structures. It is very unique that the symptoms are relieved or worsen according to the patients' position. Rupture of mature teratoma is another situation which necessitates emergent therapy. The symptoms due to rupture depends on the site, that is pericardial space, intrapulmonary, and thoracic cavity. The mechanism of rupture is suspected to autolysis, infection, and necrosis of the tumor. Anesthesia is an important issue in treating such huge mediastinal tumors, because general anesthesia using muscle relaxants can introduce acute respiratory or cardiac failure. Patient's position is again very important, for example, in the patients with orthopnea anesthesia is sometimes initiated and the patient is intubated in sitting position. Although it is rare, we must be aware of these emergent situations when we treat patients with benign mediastinal tumors. PMID- 15362561 TI - [Acute mediastinitis]. AB - Causes, diagnosis, treatment and mortality about acute mediastinitis were described. Two cases of our experience were demonstrated with chest X-ray, computed tomography (CT) and operative findings. Prevention, recognition, prompt diagnosis and early treatment of the disease were emphasized. PMID- 15362562 TI - [Traumatic injuries of the diaphragm]. AB - Traumatic injuries of the diaphragm account for only a small proportion (2.5-10%) of cases of chest trauma. However, because of the anatomical and physiological characteristics of the diaphragm, early diagnosis and treatment is necessary. The diaphragm is composed of thin, membranous muscle tissue. Since, whether a person is resting or active, the diaphragm moves in the form of a large arch in the body cavity so long as the person is alive, it differs from other parenchymatous organs, and it is difficult to detect abnormalities by diagnostic imaging by means of computed tomography (CT), etc.. Thus, the patient's general condition (especially respiratory status) upon arrival at the hospital and an analysis of fine-grained plain films of the chest and thoracoabdominal ultrasound diagnostic examinations are important. PMID- 15362563 TI - [Congenital diaphragmatic hernia]. AB - Recent progress of the treatment of congenital diaphragmatic hernia (CDH) is addressed. Early CDH onset within 24 hours of life has been a main concern for neonatologists and pediatric surgeons. Repair of CDH has changed from an emergent procedure to a delayed procedure in the last decade. Employment of new modalities including extracorporeal membrane oxygenation (ECMO), high-frequency oscillatory ventilation (HFO), inhaled NO, and surfactants is decided depending on the situation of each institution. Permissive hypercapnea/spontaneous respiration, more recent style of respiratory management aiming to avoid barotraumas, has reportedly made the outcome much better. Fetal surgery consisting of fetal tracheal occlusion and tracheal repair with ex utero intrapartum treatment (EXIT) has been applied for most serious patients with antenatal diagnosis. It is a promising approach, but recent study failed to demonstrate the improved survival or morbidity rates by tracheal occlusion of the fetuses. Other innovative approaches such as liquid ventilation, minimally invasive surgery, and lung transplantation have begun to be clinically employed. However, the impact of these approaches on the clinical results has not been determined yet. According to the report from the Japanese Society of Pediatric Surgeons, the survival rate of neonates with CDH has improved from 60% to 75% for these 10 years. PMID- 15362564 TI - [Iatrogenic perforation of the esophagus]. AB - Although advances in diagnostic methods and supportive therapy, mortality rates for esophageal perforation are still high. The most common cause of esophageal perforation is iatrogenic injury for the esophagus. In this article, causes, incidence, location of lesions, and treatment for iatrogenic perforation of the esophagus were reviewed. PMID- 15362565 TI - [Congenital esophageal atresia]. AB - Current management of congenital esophageal atresia (CEA) is described on the basis of our experience and recent literatures. Primary repair for Gross C type CEA is performed as modern standard treatment in infants without high-risk factors such as associated severe cardiac anomaly and respiratory insufficiency. Surgical strategy depends on preoperative condition of the infant therefore preoperative full assessment of the infant is very important. In general, delayed primary repair or staged repair on CEA is selected for premature infants weighing less than 1,500 g and high-risk infants. Recently, primary repair has become an effective option in premature infants without high-risk factors. In long-gap CEA, gastrostomy and/or closure of tracheoesophageal fistula is performed initially. Esophagoesophagostomy is carried out after attempts to decrease gap length. Intraoperative esophageal elongation is required in some infants. However esophageal replacement should be selected if esophageal elongation fails is impossible due to hypogenesis of lower esophagus. Thoracoscopic primary repair was recently reported as a new optional treatment. This treatment will be able to decrease the damage on the thoracic wall. However this procedure should be adopted after very careful discussion because it is difficult to accomplish without very skillful endoscopic surgical technique. PMID- 15362566 TI - Expression of HSPF1 and LIM in the lymphoblastoid cells derived from patients with bipolar disorder and schizophrenia. AB - We have previously reported the altered expressions of HSPF1 and LIM in the lymphoblastoid cell lines (LCLs) derived from Japanese patients with bipolar disorder (bipolar I disorder). The altered expression at the LCL level would be useful for developing diagnostic markers as well as a cellular model for bipolar disorder. In this study, we extended our previous study by measuring their expressions using the following samples: (1) larger number of LCLs from Japanese subjects, (2) LCLs from Caucasian subjects, and (3) LCLs from patients with bipolar II disorder or schizophrenia. We confirmed the increased expression of HSPF1 (P=0.009) and decreased expression of LIM (P=0.001) in the LCLs from patients with Japanese bipolar I disorder. These altered expressions were also observed in those from patients with Japanese bipolar II disorder (P= 0.002 for HSPF1 and P = 0.072 for LIM). We also found the altered expressions of HSPF1 in LCLs from Caucasian patients with bipolar II disorder (P=0.011) and LIM in those from patients with schizophrenia (P = 0.001). PMID- 15362567 TI - Haplotype architecture of the norepinephrine transporter gene SLC6A2 in four populations. AB - The norepinephrine transporter (NET) regulates levels of monoamine neurotransmitters integral to a variety of behaviors and autonomic functions. Two SLC6A2 polymorphisms have been used in genetic association studies, generating intriguing but nondefinitive results on traits such as hypertension and mood. One of these SLC6A2 variants is functional but rare. The other is common but not informative over the entire 48 kb SLC6A2 region and is insufficient to capture the functional diversity potentially contained within any SLC6A2 region. To elucidate SLC6A2 haplotype structure and define markers sufficient to capture haplotype diversity within detected haplotype blocks, 26 single-nucleotide polymorphisms (SNPs) were genotyped in 384 individuals evenly divided across Finnish Caucasian, US Caucasian, Plains American Indian, and African American populations. Three conserved blocks, 13.6, 12.5, and 25 kb in size and showing little evidence for historical recombination were observed in all populations. Haplotype diversity in block 1 and numbers of common haplotypes were highest in African Americans, among whom 5-6 optimal markers were sufficient to maximize diversity of each block. For other populations, 2-3 markers/block sufficed, but the optimal markers differed across populations. The SLC6A2 haplotype map and 25 marker panel (excluding the monomorphic one) is a comprehensive tool for genetic linkage studies on phenotypes related to NET function. PMID- 15362568 TI - Characterization of liver-cirrhosis nodules by analysis of gene-expression profiles and patterns of allelic loss. AB - To disclose genetic mechanisms involved in development or progression of hepatocellular carcinoma (HCC), we used a genome-wide cDNA microarray consisting of 8,448 genes to compare gene-expression profiles among 12 liver-cirrhosis nodules (LCNs) and five specimens of HCC excised from a single patient and carefully prepared by laser-capture microdissection (LCM). The expression patterns enabled us to identify 72 genes that were frequently upregulated and 57 that were downregulated specifically in the LCN specimens as compared to the HCCs. We also documented upregulation of 31 genes and downregulation of seven others in both HCC and LCN tissues. Several types of intracellular kinase, including receptor-type kinase, were upregulated in LCNs. Expression patterns of HCCs and LCNs generally represented two genetically distinct groups when subjected to a hierarchical clustering analysis, although expression profiles of two of the LCNs resembled the HCC pattern. Analysis of allelic losses at microsatellite loci revealed that LCNs showed frequent loss of heterozygosity (LOH) (33%) in chromosomal regions 6q and 22q; over half of the LCNs had lost an allele for at least one of the 28 loci examined. The presence of early genetic changes among LCNs, with additional genetic changes occurring during formation of HCCs, suggests that hepatocellular carcinogenesis follows the multistep model established for colon cancers and that some LCNs may be precancerous lesions. PMID- 15362569 TI - A clinical and genetic study in a large cohort of patients with spinocerebellar ataxia type 6. AB - In order to clarify the clinical and genetic features of SCA6, we retrospectively analyzed 140 patients. We observed an inverse correlation between the age of onset and the length of the expanded allele, and also between the age of onset and the sum of CAG repeats in the normal and the expanded alleles. The ages of onset of four homozygous patients correlated better with the sum of CAG repeats in both alleles rather than with the expanded allele calculated from heterozygous SCA6 subjects. Clinically, unsteadiness of gait was the main initial symptom, followed by vertigo and oscillopsia, and cerebellar signs were detected in nearly 100% of the patients. In contrast, extracerebellar signs were relatively mild and infrequent. The results of neuro-otological examination performed in 22 patients suggested the purely cerebellar abnormalities of ocular movements in nature. There was a close relationship between downbeat positioning nystagmus (DPN) and positioning vertigo, which became more common in the later stage. We conclude that total number of CAG repeat-units in both alleles is a good parameter for assessment of age of onset in SCA6 including homozygous patients. In addition, clinical and neuro-otological examinations suggested that SCA6 is a disease with predominantly cerebellar dysfunction. PMID- 15362570 TI - Characterization of the human nasal embryonic LHRH factor gene, NELF, and a mutation screening among 65 patients with idiopathic hypogonadotropic hypogonadism (IHH). AB - As the mouse nasal embryonic LHRH factor gene (Nelf) encodes a guidance molecule for the migration of the olfactory axon and gonadotropin-releasing hormone neurons, its human homolog, NELF, is a candidate gene for Kallmann syndrome, a disease of idiopathic hypogonadotropic hypogonadism (IHH) with anosmia or hyposmia. We report here characterization of NELF and results of mutation analysis in 65 IHH patients. Assembling EST clones, RACE, and sequencing showed that NELF mapped to 9q34.3 is composed of 16 exons and 15 introns with a 1,590-bp ORF encoding 530 amino acids. RT-PCR on a fetal brain cDNA library revealed five alternatively spliced variants. Among them, NELF-v1 has 93-94% identity at the amino acid level to mouse/rat Nelf, and four other transcripts are also highly conserved among the three species. A 3.0-kb transcript is expressed most highly in the adult and fetal brain, testis, and kidney, indicating that NELF plays a role in the function of these tissues. Mutation screening detected in a patient with IHH one novel heterozygous missense mutation (1438A>G, T480A) at the donor splice site in exon 15 of NELF. As this mutation was not found in 100 normal control individuals, T480A may be associated with IHH. Four other novel SNPs (102C > T and 1029C > T within the coding region, and two IVS14+47C > T and IVS15+41G > A) were also identified in NELF. PMID- 15362571 TI - Polymorphisms within a polymorphism: SNPs in and around a polymorphic Alu insertion in intron 44 of the human dystrophin gene. AB - A polymorphic Yb-type Alu insertion on Xp21.3 shows a genotypic gradient across worldwide populations. We used single strand conformational polymorphism (SSCP), denaturing high-pressure liquid chromatography (DHPLC), and sequencing to characterize the level of polymorphism within this region. Two novel polymorphic sites were found within the Alu insertion itself, and a further seven novel polymorphic sites in the 2-kb flanking region. Our results showed that while DHPLC was more sensitive than SSCP, the limitations of DHPLC included the lack of ability to distinguish between multiple alleles or safely identify mutations on a polymorphic background. We believe that this is the first report of polymorphic single nucleotide polymorphisms (SNPs) within a polymorphic Alu distribution and that together they promise to provide a useful marker for human population and evolutionary genetics. PMID- 15362572 TI - Dlx5, the mouse homologue of the human-imprinted DLX5 gene, is biallelically expressed in the mouse brain. AB - The mouse Dlx5 gene encodes a distal-less-related DNA-binding homeobox protein first expressed during early embryonic development in anterior regions of mouse embryo and is located on chromosome 6, which is the syntenic region to the human chromosome 7q21-q31 imprinting cluster. Recently, its human homologue, DLX5, was identified to be imprinted and maternally expressed, at least in normal human lymphoblasts and in brain tissues. In our study, we analyzed the imprinting status of mouse Dlx5 by RT-PCR, first in the F1 of a reciprocal cross between two different mouse strains, and second in heterozygous Dlx5 mutant mice. Both approaches revealed that mouse Dlx5 followed a biallelic pattern of expression in brain tissue and in testis. Our findings suggest that the Dlx5 gene escapes genomic imprinting, at least in mice of certain genetic backgrounds. PMID- 15362573 TI - A novel susceptibility locus for moyamoya disease on chromosome 8q23. AB - Moyamoya disease (MIM 252350) is characterized by stenosis or occlusion of the terminal portions of the bilateral internal carotid arteries and by abnormal vascular networks at the base of the brain. There is a high incidence of moyamoya disease in Asia, especially in Japan. Multifactorial inheritance is estimated with lambda(s)>40. Previous linkage studies have indicated that susceptibility loci for the disease are located on chromosomes 3p, 6q, and 17q. In the present study, we searched for loci linked to the disease in 12 Japanese families using 428 microsatellite markers and found significant evidence for linkage to 8q23 [maximum LOD score (MLS) of 3.6] and suggestive evidence for linkage to 12p12 (MLS=2.3). The present study revealed a novel locus for moyamoya disease. PMID- 15362574 TI - Phenotype-genotype correlation in two patients with 12q proximal deletion. AB - Proximal 12q deletion is a very rare chromosomal abnormality. Only five cases have been reported. Among the five, an Argentinian patient (Case 1) with del(12)(q11q13) and a Japanese patient (Case 2) with del(12)(q12q13.12) were analyzed because they shared several clinical features: growth and psychomotor developmental delay; strabismus; broad and short nose with anteverted nostrils; high, arched palate; large, lowset ears; widely set nipples; short fingers and clinodactyly of fifth fingers; and abnormality of the second and third toes. To clarify the correlation between the deleted genes and their phenotypes, we delimited their deleted regions by fluorescence in situ hybridization (FISH). The overlapped region in the deletions spanned 6.2 Mb where at least 15 genes were predicted to localize on the current human genome database. Among them, YAF2 and AMIGO2 were the most plausible candidates to affect growth and psychomotor retardation, respectively, in both cases. Regarding unique symptoms in each case, congenital fibrosis of the extraocular muscles found only in Case 1 may be caused by KIF21A deletion and hearing loss and cleft palate in Case 2 by COL2A1 defect. PMID- 15362575 TI - Biology and management of gastric carcinoid tumours: a review. AB - In recent times gastric carcinoid tumours have become the subject of substantial clinical interest as knowledge of their biological background and clinical importance has increased Gastric carcinoid tumours have long been considered rare lesions, amounting to less than 2% of all carcinoid tumours and less than 1% of all stomach neoplasms. New large databases suggest that the prevalence of gastric carcinoid is rising, with a true value now closer to 5% of all carcinoids. A tripartite classification system for gastric carcinoid tumours is now in common use: tumours associated with chronic atrophic gastritis; tumours associated with Zollinger-Ellison syndrome; and sporadic lesions. Gastric carcinoids associated with hypergastrinaemia are relatively benign, while sporadic lesions require aggressive surgical management. Gastric carcinoids, however, can be managed initially by endoscopic excision of accessible tumours, followed by regular endoscopic surveillance. PMID- 15362576 TI - Mesh repair of incisional hernia: comparison of laparoscopic and open repair. AB - OBJECTIVE: To compare our results of open and laparoscopic mesh repair of incisional hernias. DESIGN: Retrospective cohort study. SETTING: Teaching hospitals, The Netherlands. SUBJECTS: All patients who had had a laparoscopic (n = 25) or an open (n = 76) mesh repair of incisional hernia between January 1996 and January 2000. INTERVENTIONS: Physical examination at the time of the study. MAIN OUTCOME MEASURES: Morbidity and recurrence. RESULTS: The groups were comparable. 11 patients (14%) developed postoperative infections after open repair and 1 (4%) after laparoscopic repair (p = 0.29). Median hospital stay was 5 days (range 1-19) in the open group and 4 (range 1-11) in the laparoscopic group (p = 0.28). The 2-year cumulative incidence of recurrence was 18% after open repair (median follow-up of 17 months (range 1-46) and 15% after laparoscopic repair (median follow-up of 15 months, range 1-44). Recurrences in the laparoscopic group were all among the first 7 cases in which the mesh was fixed with staples alone. CONCLUSION: There were fewer infections and hospital stay was shorter in the laparoscopic group, but not significantly so. Recurrence rates were comparable. PMID- 15362577 TI - Preoperative routine magnetic resonance cholangiopancreatography before laparoscopic cholecystectomy: a prospective study. AB - OBJECTIVES: To assess the diagnostic value of magnetic resonance cholangiopancreatography (MRCP) in detecting common bile duct stones in the preoperative investigation of patients electively referred for gallstone disease, to find out the incidence of asymptomatic common duct stones, and to correlate clinical symptoms and history and liver function tests (LFT) with the actual occurrence of common duct stones. DESIGN: Prospective study. SETTING: General hospital, Denmark. PATIENTS: 180 consecutive non-jaundiced patients referred with symptomatic gallstones for elective cholecystectomy. INTERVENTIONS: LFT, abdominal ultrasonography, MRCP, endoscopic retrograde cholangiopancreatography (ERCP), questionnaire. MAIN OUTCOME MEASURES: Positive and negative predictive values and accuracy of MRCP, number of patients with asymptomatic stones, and correlation of symptoms with the presence of stones. RESULTS: 26/180 patients had common duct stones (14%). Only one (<1%) had an asymptomatic stone. For detection of such stones, MRCP's positive predictive value was 0.95 (95% confidence interval (CI): 0.86 to 1.00), negative predictive value 0.96 (0.93 to 0.99), and accuracy 0.85 (0.93 to 0.99). MRCP missed 5 stones 1-4 mm in size in 5 patients; 17/64 patients with raised LFTs had stones (27%). The probability of stones was highest when the patients had both raised LFTs and a dilated common (>7 mm) bile duct (82%). There were no readmissions with ductal stones in the 6-month postoperative period. CONCLUSIONS: The predicive values of MRCP were fairly good, but MRCP misses some small stones <5 mm in size. Asymptomatic stones in the common duct are not common in this population and should not be screened for. The probability of stones increases with the number of predictive factors. Patients should be questioned carefully about signs of biliary obstruction, and only be offered preoperative MRCP should they have a suspicious history, raised LFTs, or a dilated common duct. PMID- 15362578 TI - Reintervention after laparoscopic and open cholecystectomy in Sweden 1987-1995: analysis of data from a hospital discharge register. AB - OBJECTIVE: To find out the incidence of cholecystectomy and of reintervention after cholecystectomy in Sweden 1987 to 1995, and to compare mortality and reintervention after simple laparoscopic and conventional open cholecystectomy (without exploration of the common bile duct or simultaneous operation). DESIGN: Analysis of data from Swedish national registers. SETTING: Two hospitals and government department, Sweden. MAIN OUTCOME MEASURES: Mortality and reintervention during readmission within one year after cholecystectomy classified as: reoperation on bile duct, endoscopic or percutaneous reintervention, or reoperation for wound complication, bleeding, or unspecified cause. RESULTS: Incidence of cholecystectomy rose between 1987-89 and 1993-95 from 0.97 to 1.04 for men and from 1.70 to 2.05 operations/1000 inhabitants for women. Reoperation on the bile ducts declined from 1987 to 1991 but returned to previous levels thereafter. Endoscopic reinterventions increased tenfold from 1987 to 1995, whereas those for general complications and mortality did not change significantly. Among simple cholecystectomies laparoscopic surgery was associated with an increased risk of endoscopic reintervention, odds ratio 1.8 (95% CI 1.2 to 2.6), and with a lower risk for postoperative mortality, odds ratio 0.5 (95% CI 0.3 to 0.8). CONCLUSIONS: Incidence, mortality, and readmission with reintervention are important endpoints in gallbladder surgery. Significant changes in these variables were identified after the introduction of laparoscopic cholecystectomy. PMID- 15362579 TI - Symptoms and reflux competence in relation to anatomical findings at reoperation after laparoscopic total fundoplication. AB - OBJECTIVE: To investigate the mechanisms and anatomical failures after total laparoscopic fundoplication using the symptoms and findings at reoperation. DESIGN: Prospective open study. SETTING: University hospital, Sweden. PATIENTS: Twenty-one patients who were reoperated on a median of 33 (0.5-102) months after laparoscopic fundoplication. INTERVENTIONS: The patients were divided into three groups according to the mode of presentation. The first group presented with dysphagia and no gastro-oesophageal reflux (GOR) (n = 6). The second group (n = 11) had recurrent GOR and the third group (n = 4) complained of a sense of excessive fullness. RESULTS: In the dysphagia group the reason for it in 4 patients was severe fibrosis in the hiatal region including the right part of the fundoplication. One patient had correctly located fundoplication but it was too tight. In the last patient the part of the stomach used was too low down. All patients in the GOR group had a slippage and rupture of the fundoplication. Ten patients also had a recurrent hernia. In 6/11 patients the fundal mobilisation was incomplete. In the last group (excessive fullness) one patient had a postoperative leak from the fundal part, one patient a para-oesophageal hernia, and one patient an intact but herniated repair. One further patient had an intact abdominal oesophagus and crural repair, but a large portion of the stomach had herniated through the left part of the fundoplication and acted as a volvulus. CONCLUSIONS: Dysphagia was caused by hiatal fibrosis or other technical failures rather than a normal tight fundoplication. Using the wrong part of the stomach causes recurrent heartburn. The laparoscopic suturing technique must be improved. PMID- 15362580 TI - Benefit of venous resection for ductal adenocarcinoma of the pancreatic head. AB - OBJECTIVE: To find out whether there is any benefit from venous resection during pancreaticoduodenectomy for ductal pancreatic adenocarcinoma. DESIGN: Retrospective study. SETTING: University Hospital Mannheim/Heidelberg, Germany. INTERVENTIONS: 271 patients had resections for ductal adenocarcinoma of the pancreatic head between 1980 and 2001. The outcome of patients who did (n = 68) and who did not (n = 203) have simultaneous resection of major veins (portal vein and/or superior mesenteric vein) were compared. MAIN OUTCOME MEASUREMENT: 5 year survival. RESULTS: The groups differed significantly regarding stage, perineural infiltration, lymphangiosis carcinomatosa, operating time, blood loss, and blood transfusion. However, there was no difference in perioperative morbidity (27% and 22%), mortality (4% and 3%), and long-term survival (at 5 years 23% and 24%). Subgroup analysis of patients with margins free of tumour (R0 resections) showed that those patients who had venous resections in whom histological examination did not show infiltration of tumour had the most favourable outcome. CONCLUSION: There is no reason to exclude patients with suspected venous infiltration from radical pancreaticoduodenectomy including venous resection. PMID- 15362581 TI - Closure of small bowel stomas on postoperative day 10. AB - OBJECTIVE: To find out if early closure of a defunctioning small bowel stoma (day 10) was feasible and safe. DESIGN: Prospective non-randomised study. SETTING: University hospital, France. INTERVENTIONS: During a 42-month period (January 1998-June 2001), all patients with a temporary small bowel stoma were elected for early closure on postoperative day 10 in a non-randomised prospective study. The procedure was considered only if the patient was not taking steroids, was in good condition, and had not developed wound or general sepsis after the initial operation. Other patients' stomas were closed after the usually recommended delay (>8 weeks). MAIN OUTCOME MEASURES: Postoperative complications, delay to recover bowel activity, and to resume oral feeding, and duration of hospital stay. RESULTS: Thirty-six patients were included in the study: 14 patients in the early group and 22 in the delayed group. There were no postoperative deaths. Three patients developed wound abscesses, two in the early group and one in the delayed group. The median (range) duration of hospital stay was longer in the delayed group: 36 (14-84) days, than in the early group: 22 (18-29) days (p < 0.01). CONCLUSIONS: Small bowel stomas can be closed in selected healthy patients on postoperative day 10 without major complications. PMID- 15362582 TI - 25 years experience of the surgical treatment of phaeochromocytoma. AB - OBJECTIVE: To assess the outcome of the surgical treatment of patients who had adrenalectomy for phaeochromocytoma. DESIGN: Retrospective clinical study. SETTINGS: University hospital, Germany. SUBJECTS: 87 consecutive patients with phaeochromocytoma who were operated on. INTERVENTIONS: 29 flank and 58 transabdominal adrenalectomies between 1974 and 2000. RESULTS: The mean tumour diameter was 5 cm (range 2-13), and the mean weight 91 g (range 7-550). The postoperative hospital stay was 11 days. The flank incision entailed the shortest operating time (95 minutes). Two of the phaeochromocytomas were malignant. There were two wound infections but no deaths. With a correct selection of patients, a flank incision is safe. Endoscopic retroperitoneal adrenalectomies should be preferred. PMID- 15362583 TI - Injections of botulinum A toxin for the treatment of anal fissures. AB - OBJECTIVE: To find out how injections of botulinum A toxin influence the healing of anal fissures. DESIGN: Retrospective study. SETTING: Medical University of Lodz, Poland. SUBJECTS: 13 patients (6 women, 7 men), mean age 49 (range 31-78), treated with injections of botulinum A toxin 50 units on either side of the anal fissure into the internal anal sphincter from May to December 1999. MAIN OUTCOME MEASURES: Complications and relapse. RESULTS: Seven fissures had healed by one month and four by two months. Two remained unhealed but asymptomatic. There was no incontinence of flatus or faeces after three months of treatment. Resting anal pressure was significantly lower in 10 of 13 patients compared with before treatment (p < 0.05). One fissure relapsed after 4 months and this patient had a successful anal stretch. CONCLUSION: Injection of botulinum A toxin gives good results in the treatment of anal fissures. PMID- 15362584 TI - Regional differences in the use of a vascular surgical service and incidence of amputations in a well-defined geographical area. AB - OBJECTIVE: To evaluate regional differences in the use of a vascular surgical service in the treatment of critical lower limb ischaemia and incidence of amputation in the 1990s in a well-defined geographical area. DESIGN: Retrospective study. SETTING: One university and five county hospitals, Finland. SUBJECTS: All referrals to the university hospital vascular surgical unit for chronic critical lower limb ischaemia and the number of major amputations in the region. MAIN OUTCOME MEASURES: Numbers of new vascular surgical consultations and amputations in 11 municipalities. Correlation between numbers of consultations and amputations. RESULTS: Between the subregions the age-standardised incidence of new vascular surgical consultations in the 15-85 year old population varied from 52.4 to 104.7/10(5) and the incidence of amputation from 10.2 to 24.8/10(5). There was an inverse correlation between the numbers of consultations and amputations. The most significant inverse correlation was between consultations and below knee amputations in diabetic patients (r = -0.70). For above knee amputations there was no correlation (r = -0.21). CONCLUSION: An active referral policy leads to reduced amputation rates. PMID- 15362585 TI - Trauma management and education in europe: a survey of twelve geographically and socioeconomically diverse European countries. AB - OBJECTIVE: To record the current standards of management and education in trauma surgery in 12 geographically and socioeconomically diverse countries in Europe. DESIGN: Questionnaire study. SETTING: Teaching hospital, Austria. INTERVENTION: Questionnaire sent to experts on trauma in Austria, France, Germany, Italy, The Netherlands, Norway, Portugal, Romania, Spain, Sweden, Turkey, and the United Kingdom. MAIN OUTCOME MEASURE: Comparison of management of patients before, during, and after admission to hospital, and opportunities for initial and in service training. RESULTS: Management of patients and opportunities for training varied considerably from country to country, ranging from an organised trauma service throughout with specialised training to a haphazard and variable service that depended more on individual hospitals, doctors and patients. CONCLUSIONS: Standardisation of management and training would be desirable, and should be possible at least in countries that are members of the European Union. PMID- 15362586 TI - Recording of postoperative complications: quantity and quality. AB - OBJECTIVE: To evaluate a simple working procedure for recording postoperative complications. DESIGN: Prospective cohort study. SETTING: Teaching hospital, Denmark. SUBJECTS: 218 of 312 patients who were operated on from March to July 1996. INTERVENTIONS: Follow up 30 days after operation by looking up the local register of complications, review of medical records and by structured telephone interview with the patient. MAIN OUTCOME MEASURES: Morbidity. RESULTS: All the patients were recorded in the local register, showing a complication rate of 21% (n = 49). However the medical record and the telephone interview showed complication rates of 68 (31%) and 76 (33%), respectively. CONCLUSION: It is possible to improve the quantity of the recording by a simplified working procedure, while the quality is still a challenge. PMID- 15362587 TI - Pedunculated lipoma of the oesophagus in a patient with achalasia. PMID- 15362588 TI - Castleman disease in differential diagnosis of a pancreatic mass. PMID- 15362589 TI - Giant mediastinal parathyroid adenoma presenting with a hyperparathyroid crisis and leading to postoperative hungry bone syndrome. PMID- 15362590 TI - Position paper: cathartics. AB - The administration of a cathartic alone has no role in the management of the poisoned patient and is not recommended as a method of gut decontamination. Experimental data are conflicting regarding the use of cathartics in combination with activated charcoal. No clinical studies have been published to investigate the ability of a cathartic, with or without activated charcoal, to reduce the bioavailability of drugs or to improve the outcome of poisoned patients. Based on available data, the routine use of a cathartic in combination with activated charcoal is not endorsed. If a cathartic is used, it should be limited to a single dose in order to minimize adverse effects of the cathartic. A review of the literature since the preparation of the 1997 Cathartics Position Statement revealed no new evidence that would require a revision of the conclusions of the Statement. PMID- 15362591 TI - Treatment of severe pediatric ethylene glycol intoxication without hemodialysis. AB - BACKGROUND: There is limited experience treating severe ethylene glycol poisoning in children without hemodialysis. The objective of this study was to describe the clinical course and outcome of severe pediatric ethylene glycol poisoning treated without hemodialysis. METHODS: Patient records were identified retrospectively by hospital discharge diagnosis (ICD-9 code) of ethylene glycol poisoning from 1999 through 2002 at a pediatric medial center. Patients with initial serum ethylene glycol concentrations less than 50 mg/dL or those who received hemodialysis were excluded. RESULTS: Six patients with an age range of 22 months to 14 years were admitted for treatment of ethylene glycol poisoning over a four-year period. Initial serum ethylene glycol concentrations ranged from 62 to 304 mg/dL (mean 174.0 mg/dL). The lowest-measured individual serum bicarbonates ranged from 4 to 17 mEq/L. All patients were initially admitted to intensive care. One patient received ethanol only, two patients received fomepizole only, and three patients received a loading dose of ethanol and then were converted to fomepizole therapy. None of the patients received hemodialysis. Treatment was continued until the serum ethylene glycol was less than 10 mg/dL. Metabolic acidosis resolved with intravenous fluid and supplemental bicarbonate within 24h. All patients had a normal creatinine upon presentation and at discharge. The mean length of stay in intensive care was 21h and on the ward was 33.7h. One episode of hypoglycemia occurred in a 22-month-old. All patients recovered without evidence of renal insufficiency or other major complications at discharge. CONCLUSION: Six pediatric patients with severe ethylene glycol intoxication and normal renal function were successfully treated without hemodialysis. PMID- 15362592 TI - Dipyrone overdose. AB - BACKGROUND: Dipyrone is a pyrazolone derivative used as an analgesic and antipyretic. Agranulocytosis, dipyrone's most serious and potentially fatal adverse effect, has led to its withdrawal in several countries. However, agranulocytosis is subject to geographical variability, ratio with at risks ranging from 0.8-23.7. In many countries dipyrone is still widely used in adults and children and even as an over-the-counter (OTC) preparation. Information on the effects of dipyrone overdose is scanty. OBJECTIVE: To determine the demographic and clinical characteristics of dipyrone overdose. METHODS: Retrospective review of prospectively collected poison center data on acute exposure to dipyrone over a three-year period. The data were subjected to descriptive analysis. Mann-Whitney test and Chi-square analysis were performed where relevant. RESULTS: A total of 243 records met the inclusion and exclusion criteria. Median age was 17y (4m-83y), median amount 5 g (250 mg-45 g), and median time to consultation was 2 h (5 min-48 h). Toxic events (49) occurred in 39 (16%) patients; 57% of these were gastrointestinal and all were mild. Time to consultation was longer in the symptomatic patients (4 h vs. 1.5 h, respectively, p=0.001) and in children (8 h vs. 3.5 h in adults). Suicidal patients ingested significantly larger amounts (8 g vs. 3.7 g, respectively, p=0.001), as did patients with gastrointestinal symptomatology (7.5 g vs. 5 g in asymptomatics, p=0.001). No agranulocytosis was reported. DISCUSSION: Dipyrone overdose is associated with mild, mainly gastrointestinal toxicity; this was noted at a median dose of 7.5 g. Early gastrointestinal decontamination may have prevented toxicity. The suggested treatment includes gastrointestinal decontamination (if <1 h since ingestion) and supportive measures. PMID- 15362593 TI - Effect of anticholinergic drugs on the efficacy of activated charcoal. AB - BACKGROUND: Although it is a commonly held belief that the ingestion of drugs with an anticholinergic action would prolong the duration of time after drug ingestion for effective gastrointestinal decontamination, data are lacking to support this belief. The purpose of this study is to determine whether activated charcoal is more effective in the presence of concurrent anticholinergic activity. METHODS: A three-limbed randomized crossover study in 10 healthy volunteers was completed to determine the ability of a 50 g dose of activated charcoal to reduce the bioavailability of a simulated overdose of acetaminophen (12 x 325 mg tablets) in the presence and absence of a concurrently present anticholinergic drug, atropine (0.01 mg/kg I. M. administered 15 min prior to the acetaminophen ingestion). RESULTS: After the acetaminophen ingestion, median Cmax occurred at 1 h for all three exposures but was lower in the atropine-treated study arm (31+/-19 mg/L) than in the control or charcoal alone intervention arms (49+/-13 and 51+/-16 mg/L, respectively) (P<0.05). Compared to the control area under the serum concentration vs. time curve, a single dose of activated charcoal 1 h after drug ingestion reduced acetaminophen bioavailability by 20% (95% CI 4 36%) and by 47% (95% CI 35-59%) in the presence of atropine (P<0.05 atropine plus charcoal vs. charcoal alone). CONCLUSIONS: Our data support the belief that activated charcoal is more effective in the presence of anticholinergic activity. Additional study is required to determine whether in patients with anticholinergic drug overdose, activated charcoal is effective at times beyond the recommendation for overdoses of drugs without this pharmacodynamic effect. PMID- 15362594 TI - Frequency and nature of recorded childhood immunization-related errors in Greece. AB - BACKGROUND: While routine immunizations are very safe, their administration to healthy children requires minimization of immunization programmatic errors. In order to estimate the incidence and ascertain the nature of reported immunization errors in the Greek childhood population, we have undertaken a study using data from the National Poison Information Center in Greece, which also has the responsibility to address medication-induced errors. METHODS: All immunization errors concerning children and reported to the National Poison Information Center during the 2-yr period 1999-2000 were retrieved and the conditions of their occurrence were examined. The incidence of reported errors was calculated under the assumption that during each year 100,000 children are born in Greece, and during their childhood they receive a total of about 20 immunization doses of all childhood immunizations. RESULTS: There were 40 immunization errors reported, corresponding to a reported incidence of about 11 per million immunization doses. Of these errors, 20 concerned OPV, 13 DTP, 5 MMR, 1 Haemophilus influenza and 1 Hepatitis B immunizations. In 12 instances an erroneous route was used (out of which 11 concerned OPV), whereas overdose was documented in 13 instances (out of which 8 concerned OPV). The third most common error was administration of DTP instead of the recommended Td vaccine. No adverse patient outcomes were reported. CONCLUSIONS: In Greece, reported errors in immunization practice are relatively rare. Packaging modifications (about one in three errors in this study) of the OPV and DTP could further reduce their incidence. PMID- 15362595 TI - Relative toxicity of selective serotonin reuptake inhibitors (SSRIs) in overdose. AB - BACKGROUND: Selective serotonin reuptake inhibitors (SSRIs) have increasingly replaced tricyclic antidepressants (TCAs) in the treatment of depression. They appear to be safer in overdose, but there is little information on their spectrum of toxicity in overdose, or relative toxicity of each agent. OBJECTIVE: To determine the effect of SSRIs in overdose, as a group, and the relative toxicity of five different SSRIs. METHODS: A review of consecutive SSRI poisoning admissions to a single toxicology unit. Outcomes examined were length of stay [LOS], intensive care [ICU] admission rate, coma, seizures, electrocardiographic [ECG] abnormalities, and presence of serotonin syndrome [SS]. Logistic regression was used to model the outcome QTc >440 msec. RESULTS: There were 469 SSRI poisoning admissions analyzed after exclusions. The median LOS for all SSRI overdose admissions was 15.3 h (IQR: 10.5-21.3) and 30 of 469 (6.4%; 95% CI 4.3 9.0%) cases were admitted to ICU. The incidence of seizures was 1.9% and coma was 2.4%. Serotonin syndrome occurred in 14% of overdoses. Comparison of median QTc intervals of the five SSRIs was significantly different (p=0.0002); citalopram (450 IQR: 436-484) was individually different to fluoxetine (p=0.045), fluvoxamine (p=0.022), paroxetine (p=0.0002), and sertraline (p=0.001). The proportion of citalopram overdoses with a QTc >440 msec was 68%, differing significantly from sertraline (adjusted OR: 5.11 95% CI 2.32-11.27). Comparison of median QT intervals of the five SSRIs was statistically different (p=0.026); citalopram (400 IQR: 380-440) was individually different from sertraline (p=0.023). CONCLUSIONS: This study shows SSRIs are relatively safe in overdose despite serotonin syndrome being common. The exception was citalopram, which was significantly associated with QTc prolongation. We believe that cardiac monitoring should be considered in citalopram overdose, particularly with large ingestions and patients with associated cardiac disease. PMID- 15362596 TI - Lead-contaminated drinking waters in the public schools of Philadelphia. AB - Lead exposure is a preventable environmental health concern. Young children between the ages of 1 to 6 are most susceptible to its clinical effects. This article reports the results of lead level determinations in the drinking water of Philadelphia's public school buildings and remediation efforts aimed at dealing with this public health concern. METHODS: Water samples were collected from drinking sources in 292 school buildings in Philadelphia from May 2000 through January 2001. These samples were collected and sent to reference laboratories for determination of lead levels. RESULTS: A total of 42.5% (124) of schools had water lead levels not exceeding the action level of 20 ppb, of which 3.1% had nondetectable levels or levels less than 5 ppb. A total of 28.7% of buildings had water lead levels ranging from 20 to 50 ppb, 11.6% had levels between 50-100 ppb, and 17.1% had water lead levels of 100 ppb or more. CONCLUSION: A total of 57.4% of Philadelphia's public school buildings had water lead levels exceeding the Environmental Protection Agencies (EPA) action level of 20 ppb, and 28.7% of school buildings had water with mean lead levels in excess of 50 ppb. Depending on the volume of water consumed, drinking water from school buildings may be a significant source of lead exposure for children in their formative years of development. Although Philadelphia's public school buildings were evaluated, lead contaminated drinking water in schools is not only an urban concern. School buildings in suburban and rural areas may have similar water lead levels, and testing programs are desirable. PMID- 15362597 TI - Refractory hypoglycemia from ciprofloxacin and glyburide interaction. AB - Patients taking multiple medications may suffer from unpredictable and complex drug-drug interactions resulting in significant morbidity and mortality. There are few reports in the literature of hypoglycemia with concurrent administration of an oral hyperglycemic agent and a fluoroquinolone antibiotic. We present a case of a diabetic patient taking glyburide who was prescribed ciprofloxacin and developed prolonged hypoglycemia, which persisted for over 24 hours. The mechanisms by which these agents interact to produce prolonged hypoglycemia are complex and probably multifactorial. Patients stabilized on glyburide who are started on a fluoroquinolone should have their glucose levels monitored closely. PMID- 15362598 TI - Toxicity following unintentional DDT ingestion. AB - INTRODUCTION: Dichlorodiphenyltrichloroethane (DDT) ingestion is an uncommon cause of poisoning worldwide. To date, no cases of renal impairment after oral intake of DDT in humans have been reported. We describe the clinical course and management of two patients presenting after DDT ingestion, one of whom developed acute oliguric renal failure. CASE REPORT: A father and son mistook DDT powder for flour while preparing fish for a meal, and after eating they developed symptoms compatible with acute organochlorine insecticide poisoning. Both were intubated endotracheally due to recurrent convulsions and loss of consciousness followed by admission to the intensive care unit. Both cases developed severe metabolic acidosis. Acute oliguric renal failure (ARF) was diagnosed in the son in the second day, with a blood urea nitrogen level of 47 mg/dl and creatinine 6.4 mg/dl. Urinalysis disclosed abundant RBCs on the third day. Vigorous fluid resuscitation and strict monitoring helped reverse its clinical course by the tenth day. Both patients recovered within two weeks and were discharged without sequelae. CONCLUSION: Clinicians should not overlook the possibility of DDT poisoning in the differential diagnosis of acute renal failure and seizures. More strict measures should be taken to prohibit misidentification of DDT and similar products, particularly in the developing world. PMID- 15362599 TI - Acute renal failure following ingestion of manganese-containing fertilizer. AB - Fertilizers are used to promote the survival and growth of plants and crops and have a good safety record when used properly. The basic elements in fertilizer include phosphorus, nitrite, and potassium. In addition, there are additive agents that vary for different crops and which may include some metals. Acute intoxication by ingesting fertilizer includes damage to various organ systems as well as severe cardiovascular or respiratory distress. We report the case of a 64 year-old man who ingested about 700 mL of fertilizer and suffered acute renal failure, hyperkalemia, and mild methemoglobinemia. After supportive care and emergent hemodialysis for hemodynamic instability due to hyperkalemia, the renal function of the patient recovered in four days. PMID- 15362600 TI - Oral sumatriptan-induced myocardial infarction. AB - BACKGROUND: Sumatriptan has been used in the treatment of migraine and other vascular headaches since 1993 in the United States. Its side effects include chest pains in 3% to 8% of patients who have known cardiac risk factors. This is a case report of a 45-year-old woman with no history of cardiac risk factors who had a myocardial infarction after her monthly dose of oral sumatriptan. METHODS: The patient was examined in the emergency room, evaluated by electrocardiography, and serial evaluations of cardiac enzymes over the next 24 h. She was admitted to the cardiology ward. A cardiac catherization and additional laboratory studies were performed the following day. RESULTS: The catherization revealed normal heart function, but a 60% to 70% non-flowing stenosis within the first septal perforator. Laboratory indices for cardiac risk were within normal ranges. CONCLUSIONS: Patients without cardiac risk factors may experience myocardial infarction following an oral dose of sumatriptan. PMID- 15362601 TI - Aphrodisiac drug-induced hemolysis. AB - Volatile alkyl nitrites have been used during the past decades for "recreational purposes," and for intensifying sexual experience. Their use has been associated with methemoglobinemia and hemolysis. We report three patients who presented over the past year with acute hemolysis after inhalation of butyl nitrite, two of them had glucose-6-phosphate dehydrogenase (G6PD) deficiency. PMID- 15362602 TI - Skin toxicity from glyphosate-surfactant formulation. AB - Glyphosate (N-[phosphonomethyl]glycine) is a nonselective herbicide used in agriculture as a foliage spray for the control and the destruction of herbaceous plants. Adverse skin reactions due to contact with this compound have been rarely described. We report a case of a 78-year-old woman presenting with extensive chemical burns on her trunk and legs caused by accidental contact with a glyphosate-surfactant formulation. The lesions healed in four weeks without scarring. PMID- 15362603 TI - Effectiveness of delayed use of crotalidae polyvalent immune Fab (ovine) antivenom. AB - Traditionally, horse-serum-based antivenom has been used in the United States for North American crotaline snake evenomation. Crotalidae polyvalent immune Fab (ovine) was approved in 2000 for use in mild to moderate envenomations. The manufacture recommends use within 6 h of envenomation. Published postmarketing retrospective reports describe its use up to 9 h after envenomation. We describe a case of effective use of FabAV 52 h after envenomation with resultant correction of coagulopathy and mild improvement of local symptoms. PMID- 15362604 TI - Nephrotoxic effects of X-ray contrast media. AB - The annual sale of x-ray contrast media (CM) now represents 60 million doses, and contrast nephropathy (CN) has been the third-leading cause of hospital-acquired acute renal failure. In this review article, physicochemical, pharmacokinetic, and pharmacodynamic properties of CM are surveyed. The definition of CN is presented, as well as the mechanisms involved in the pathogenesis. Low osmolar monomeric CM (LOCM) are less nephrotoxic than the older ionic high osmolar CM (HOCM), but in risk patients the incidence of CN is still high after intravascular administration of LOCM. Non-ionic dimeric CM are iso-osmolar to plasma (IOCM), and they have reduced the nephrotoxicity even more than LOCM. The most important risk factors for CN are diabetes mellitus and impaired renal function. Selection of patients, hydration, and type of CM are essential for prevention and prophylaxis of CN. We do not recommend routine prophylaxis with N acetylcysteine (NAC) during CM investigations, but its use in high-risk patients should be considered. PMID- 15362605 TI - The benign clinical course following a large pediatric montelukast ingestion. PMID- 15362606 TI - Successful use of hemodialysis in acute valproic acid intoxication. PMID- 15362607 TI - [Niche characteristics of plant populations in Quercus aliena var. acuteserrata stands in Qinling Mountains]. AB - Based on the data collected from 31 plots and using Levins, Hurlbert and Pianka formulas, this paper calculated and analyzed the niche breadths and overlaps of 24 tree and 29 shrub populations in Quercus aliena var. acuteserrata stands in Mt. Qinling, Shaanxi. The results showed that the Levins and Hurlbert's niche breadths of Quercus aliena var. acuteserrata , Pinus armandi, Toxicodendron vernicifluum, Populus davidiana and Pinus tabulaeformis in arbor layer were 3.695, 1.695, 1.325, 0.840, 0.702 and 0.036, 0.299, 0.568, 1.721, 2.701 respectively, and those of Corrylus heterophylla, Lespedeza formosa and Smilax vaginata in shrub layer were 1.833, 1.466, 0.984 and 0.111, 0.300, 0.594, respectively. In general, the niche overlaps between the species with bigger niche breadth and other species were higher, and those of the species having similar environmental requirements were also higher. PMID- 15362608 TI - [Water conservation functions of several artificial forest ecosystems in semiarid region of western Liaoning Province]. AB - Five artificial forest ecosystems in semiarid region of western Liaoning Province were selected to test their water conservation capacity. The average interception rate of different artificial forest ecosystems varied from 14.58% to 37.19%, and the order was H. rhamnoides forest ecosystem > P. tabulaeformis-H. rhamnoides mixed forest ecosystem > P. simonii-H. rhamnoides mixed forest ecosystem > P. tabulaeformis forest ecosystem > P. simonii forest ecosystem. The thickness of the litter layer in different forest ecosystems varied from 1.6 to 4.1 cm, and the biomass of the litter varied from 1890.4 to 6425.2 kg x hm(-2). The order of the thickness and the biomass of the litter in different forest ecosystems was H. rhamnoides forest ecosystem > P. tabulaeformis-H. rhamnoides mixed forest ecosystem > P. simonii-H. rhamnoides mixed forest ecosystem > P. tabulaeformis forest ecosystem > P. simonii forest ecosystem. The maximum water holding capacity of the litter in different forest ecosystems varied form 5957.7 to 19332.9 kg x hm(-2), and the order was H. rhamnoides forest ecosystem > P. tabulaeformis-H. rhamnoides mixed forest ecosystem > P. simonii-H. rhamnoides mixed forest ecosystem > P. tabulaeformis forest ecosystem > P. simonii forest ecosystem. The water holding capacity of non-capillary porosity of 0-40 cm soil layer in different forest ecosystems varied from 23.70 to 37.85 mm, and the order was H. rhamnoides forest ecosystem > P. simonii-H. rhamnoides mixed forest ecosystem > P. tabulaeformis-H. rhamnoides mixed forest ecosystem > P. simonii forest ecosystem > P. tabulaeformis forest ecosystem. Among the five artificial forest ecosystems, H. rhamnoides forest ecosystem had the best water conservation capacity, and the mixed forest ecosystems had a better water conservation capacity than P. tabulaeformis and P. simonii forest ecosystem. PMID- 15362609 TI - [Dynamics and species-diversities of artificial Sonneratia apetala, Sonneratia caseolaris and Kandelia candel communities]. AB - Studies on the dynamics and species-diversities of artificial Sonneratia apetala, Sonneratia caseolaris and Kandelia candel communities showed that the arbor layer of S. apetala and S. caseolaris communities contained two distinctive sub-layers. The upper layer was composed of S. apetala or S. caseolaris, and the medium layer was composed of K. candel and Aegiceras corniculatum. The dominant population S. apetala or S. caseolaris in S. apetala and S. caseolaris communities only had old aged individuals but no regenerations appeared recently, while K. candel and A. corniculatum were actively progressive populations, which would possibly become dominant populations during the course of community development, showing that S. apetala and S. caseolaris were pioneer species for plantation, which could promote natural colonization of regional mangrove species when planted at open mudflat. K. candel was the actively progressive population in K. candel community, its natural regeneration and succession could be successful, while A. corniculatum and B. sexangula were initially progressive populations. The species composition and species-diversities of S. apetala and S. caseolaris communities were similar, both contained the main species in K. candel community, namely, K. candel, A. corniculatum and Bruguiera sexangula, showing that S. apetala and S. caseolaris could co-exist with these regional species. To introduce and plant them could help to form complicated and diversified mangrove communities. During the early stage of the development, the species-diversities of S. apetala and S. caseolaris communities were higher when their planting density was high. The species-diversities grew slightly higher when the communities became older. PMID- 15362610 TI - [Radiation characteristics in a tropical seasonal rain forest canopy gap]. AB - Based on the observations in the radiation with different wavelengths in tropical seasonal rain forest canopy gap, interior of forest, and meteorological station in different seasons, the radiation characteristics of canopy gap were discussed. The results showed that the value of different wavelength radiation had the phenomenon of "suddenly emergent" at the center and north edge of canopy gap. Moreover, each radiation value reached the maximum rapidly, and then decreased quickly around the noon. The daily gross radiation of different wavelengths within the canopy gap was bigger than that in the interior of forest, but smaller than that at meteorological station. Compared to the north edge, the daily gross radiation in the center of the gap was bigger in dry-hot season, rain season and later rain season, but smaller in fog-cool season, which was influenced by dense fog, solar angle and azimuth. The total daily gross solar radiation at the center was the maximum when the solar angle was the biggest in rain season, higher in later rain season and dry-hot season, and was the minimum in fog-cool season. The ratios of infrared and visible light radiation to total solar radiation varied with the seasons at the center and north edge of the canopy gap, which fully indicated the environmental heterogeneity within the canopy gap. Compared with meteorological station and interior of forest, the ratios of infrared radiation to total solar radiation in the gap were bigger than those of meteorological station, but smaller than those of interior of forest, which were just contrary to the ratios of visible light to total solar radiation. In addition, the seasonal variation of the ratios of visible light to total solar radiation was higher than that of the ratios of infrared radiation to total solar radiation within the gap. Generally, canopy gap could increase infrared radiation and decrease visible light radiation in tropical seasonal rainforest. PMID- 15362611 TI - [Effect of phosphorus stress on chlorophyll biosynthesis, photosynthesis and biomass partitioning pattern of Fraxinus mandchurica seedlings]. AB - Phosphorus is essential to forest trees for their growth and development, because of its important role in many metabolic processes of trees. But, most part of phosphorus in soils of Northeastern China is immobile and unavailable, and thus, many forest trees often suffer from phosphorus stress which affects the biomass accumulation of forest trees. The aim of this study is to understand how the chlorophyll biosynthesis, photosynthesis and biomass partitioning of Fraxinus mandchurica seedlings response to phosphorus stress. Sand culture experiment showed that when the seedlings were under phosphorus stress, their 5 aminolevulinic acid (ALA) biosynthesis rate and porphobilinogen (PBG) synthase activity were reduced, but no significant changes in chlorophyll a and b, total chlorophyll and carotenoid contents were found. The nitrogen content in leaves and roots and the phosphorus content in all parts of seedlings were reduced under phosphorus stress, and the reduction of N content in leaves induced the decrease of maximum quantum efficiency of PS II (Fv/Fm) efficiency of PS II photochemistry (phi(PS II)), electron transport rate (E(TR)), and maximum photosynthesis rate (Amax). The effect of phosphorus stress on chlorophyll biosynthesis didn't affect photosynthesis. Under phosphorus stress, the total biomass decreased, but more biomass was partitioned into roots. PMID- 15362612 TI - [Snow/wind damage in natural secondary forests in Liaodong mountainous regions of Liaoning Province]. AB - In early spring of 2003, the secondary forests at the Qingyuan Experimental Forests (QEF) of the Institute of Applied Ecology, Chinese Academy of Sciences suffered from the damage caused by snow and wind (snow/wind damage). This damage occurred at 800 m above sea level was the most serious one since 1949. In order to make clear its occurrence and process and to analyze its possible influences on the secondary forest system, comprehensive investigations were conducted soon after the snow/wind damage. The results showed that the occurrence of this damage was due to the special site conditions (higher sea level and steeper slopes) and air temperature, which suited the wet snow formation and the snow accumulation on trees after a large scale of precipitation. The more seriously damaged areas were those with the stands of Acer mono, Juglans mandshurica, Populus spp. and so on. The ratio of damaged trees was negatively correlated with stand density, soil depth and size class of diameter, and positively correlated with tree height. Based on these results, the likely influences of snow/wind damage, i. e., the happening of diseases and insect pests, the vegetation changing under forests, and the factors of habitat and dominant species in natural secondary forests were also discussed. It is significant that to do observation and basic research on damaged forests will provide reasonable strategies for management of natural secondary forests. PMID- 15362613 TI - [Spatial pattern of two dominant shrub populations at transitional zone between oasis and desert of Heihe River Basin]. AB - Based on the field investigation in a 500 m x 500 m plot and applying geo statistical theory and methods, this paper described the spatial pattern of the coverage, density, height and volume of two shrub populations Nitraria sphaerocarpa and Reaumuria soongorica at the transitional zone between oasis and desert of Heihe River Basin. The average coverage of the two populations was 1.56 +/- 2.34% and 0.23 +/- 0.22%, and their average density was 1.5 +/- 1.8 clump x 100 m(-2) and 2.3 +/- 2.1 individual x 100 m(-2), respectively. The height of N. sphaerocarpa was 0.18 +/- 0.15 m, and its volume 0.10 +/- 0.18 m3. N. sphaerocarpa population exhibited strip distribution pattern, while R. soongorica population showed uniform distribution pattern. The nugget (c0)/sill (c0 + c) ratio of the semivariogram of the two populations was 0.08-0.12, which indicates that the random variance of spatial heterogeneity occupied 8%-12%. In semivariogram model, the range value (A0) of the coverage and density was 14.7 and 33.9 m for N. sphaerocarpa, and 25.2 m and 24.9 m for R. soongorica, respectively. As a whole, the random spatial heterogeneity existed at the scale of 10 m, and autocorrelated spatial heterogeneity existed at the scale of 34 m. The spatial heterogeneity of the coverage and density of the two dominant populations showed both isotropy and anisotropy, and the isotropy scope of R. soongorica population was larger than that of N. sphaerocarpa. The spatial pattern of N. sphaerocarpa population was controlled by two factors. The first was precipitation and surface runoff, and the second was oasisofication and desertification. PMID- 15362614 TI - [Effect of land use change on spatial distribution pattern of soil total nitrogen in Keerqin sandy land]. AB - By using statistics and geostatistics, this paper studied the content and spatial pattern of soil total nitrogen (TN) in a glassland and an arable land of southeast Keerqin sandy land. The TN content in 0-10 cm soil layer of the arable land reclaimed from a glassland 8 years ago was not different from that of the virgin grassland, but the TN content in 10-20 cm soil layer was higher than that in the grassland (P < 0.05). The soil TN storage in the grassland was 262.36 g x hm(-2) higher than that in the arable land, and its spatial pattern was significantly different between arable land and grassland. The heterogeneity and spatial dependence of TN in each soil layer (0-10 cm and 10-20 cm) of the arable land were larger than those of the grassland, and the ranges and fractal dimensions of TN in each soil layer of the arable land were larger than those of the grassland. There existed a clear horizontal spatial structure heterogeneity and spatial distribution pattern in the arable land, while in the grassland, the horizontal spatial structure heterogeneity was very low, and the horizontal spatial distribution pattern was random and uniform. The TN content in the 0-10 cm and 10-20 cm soil layers of the arable land had no difference (P < 0.05), its spatial structure characters had some difference (P < 0.05), and its spatial distribution pattern was very similar (r = 0.683, P < 0.05). The TN content and its spatial structure characters between two soil layers of the grassland had remarkable difference (P < 0.05), but its spatial distribution pattern in the two layers was relatively similar (r = 0.395, P < 0.05). In conclusion, agricultural activity could not only influence soil TN content, but also influence its spatial structure characteristics and spatial distribution pattern. Therefore, our study might promote the further study on the effect of land use change on soil nitrogen distribution pattern and its cycling in ecosystems. PMID- 15362615 TI - [Effect of cone-picking on natural regeneration of Korean pine in Changbai Mountain Nature Reserve]. AB - The storage and distribution of Korean pine seeds on Korean pine forest floor in Changbai Mountain Nature Reserve were investigated, and the data were compared with those in 1970s when the forest was little disturbed. Based on the dependence of natural regeneration of Korean pine on seed predating animals, the effect of commercial cone-picking on natural regeneration of Korean pine was discussed. The results showed that Korean pine seeds were generally berried under litter layer, and most of them were dropped during cone-picking. The density of Korean pine seeds was 1582-2640 x hm(-2), accounted for 0.3%-0.5% of that in 1970s. Furthermore, over 67.8% of these seeds were rotten. Unlike the seeds distributed by squirrel, many (46.7%-77.1%) of them distributed singly. Animals that pick seeds from cone and thus are important for natural regeneration of Korean pine have reduced their population or even disappeared in recent years because most Korean pine cone were collected by people. Therefore, cone-picking was one of the obstacles for natural regeneration of Korean pine in Changbai Mountain Nature Reserve. PMID- 15362616 TI - [Community structure organization and its relation with environmental factors of sown grassland in South China]. AB - Contribution of environmental factors, spatial factors, biological and random factors to community structure within a man-made grassland in southern terra was partitioned using canonical correspondence analysis. Under mowing and grazing, environmental factors explained (27.4% and 25.9%) variation of community structure. Contribution of spatial factors was 32.5%, 24.2%, respectively. Contribution of biological and random factors was 48% and 56%, respectively. Gradient in community structure and species abundance is evident along environmental gradient in mowing grassland. The environment-community/species relations were more complex in grazed grassland. Patches under different grazing intensity develop toward different directions. PMID- 15362617 TI - [Amelioration of secondary bare alkali-saline patches in Songnen Plain through inserting cornstalk]. AB - Based on the field experiment on Songnen grassland, a new method was established to ameliorate the secondary bare alkali-saline patches (SAP) through inserting cornstalk. The experiment was rested on the assumption that through inserting cornstalk in the secondary bare alkali-saline patches (SAP) to retain seeds moving over its surface, the necessary seed source could be gained; and these seeds should be able to germinate and survive successfully on the cornstalk itself or in its neighborhood, where should be more fit to grow than other sites in SAP, due to the decomposition of cornstalk and its special role, so that, the aim to restore vegetation of SAP could be achieved at a pretty low cost and rapid speed. The results showed that the seed bank in soil was increased significantly, owing to the inserted cornstalk and its operating processes. The seed number in ameliorated soil was 4020.0 +/- 1773.6 seeds x m(-2), while that in the secondary bare alkali-saline patches (SAP) was only 10.0 +/- 31.6 seeds x m(-2). Although the soil chemical and physical characters in ameliorated zone were improved to some extent, the overall situation of soil was still bad for plant growth, as the pH, soluble saline ion and organic matter were concerned. Most of Chloris virgata grew around or on the cornstalk, the plants around each cornstalk being 3.9 +/- 2.2, and the total being 48.64 +/- 38.72 g x m(-2). Therefore, this method demanded a few resources, and needed simple technology and low cost, which is potentially deserved to popularize. PMID- 15362618 TI - [Consistency between neutral landscape models and real landscape]. AB - Neutral landscape models (NLM) can provide standards against real landscapes, and are used to describe landscape pattern and process in the past few decades. In this paper, the neutral landscape models RULE and SimMap were tested against a real landscape, and a set of landscape metrics were used to quantify the spatial characteristics of real and simulated patterns. Measurements of some metrics (total number of patches, total perimeter, average patch area, aggregation index, contagion and lacunarity) suggested that definite level of consistency between NLM-generated maps and real landscape did exist at landscape or class levels. But, there were also some metrics, such as corrected patch perimeter-area ratio, fractal double-logged and edge distribution evenness, which didn't show any agreement between the generated maps and real landscape. In all, each NLM had its own strength in representing real landscape, but none of them was perfect. PMID- 15362619 TI - [Effect of soil preparation and fertilization on foliage and shoot growth of Ginkgo biloba and its medicine content]. AB - The study showed that the efficiency of various treatments in improving the height growth of Ginkgo biloba was organic fertilizer plus intercropping > soil preparation by blasting plus intercropping > organic fertilizer > soil preparation by blasting > intercropping, and the height growth increased by 14.5%, 8.6%, 5.7%, 3.2% and 0, respectively. The efficiency of the treatments in improving new shoot growth was organic fertilizer plus intercropping > soil preparation by blasting plus intercropping > intercropping > organic fertilizer > soil preparation by blasting, and the new shoot growth increased by 58.1%, 36.6%, 33.1%, 30.2% and 14.0%, respectively. Soil preparation, organic fertilization and intercropping had no different effect on the number of long shoots, but their effect on the numbers of short shoots and leaves was significantly different. The most efficient treatment in improving the medicine content was organic fertilization plus intercropping. Compared with control, the content of quercetin and rutin in Ginkgo biloba leaves increased by 420% and 220%, respectively. PMID- 15362620 TI - [Nitrogen use efficiency and its relationship with nitrogen nutrition characteristics of wheat varieties]. AB - Forty wheat varieties were tested at two sites (Xuzhou and Nanjing) to evaluate genotypic and environmental variation of nitrogen use efficiency (NUE) and its relationship with nitrogen nutrition characteristics. It was shown that genotype had significant effect on NUE, but environment had less effect on NRE and NFE. All of the nitrogen nutrition traits were significantly influenced by the interaction of genotype x environment. NFE, as an integrated index reflecting the differences of grain yield and nitrogen utilization among different wheat varieties, was significantly correlated with NRE, NPP and NPS, which indicated that it should be more useful in evaluating nitrogen use efficiency. NFE could be improved by increasing N assimilation and translocation after anthesis to effectively increase the N utilization of wheat. PMID- 15362621 TI - [Soil arthropod communities in different successional tropical secondary forests in Xishuangbanna]. AB - Studies on the soil arthropod community structure and its seasonal variation in four types of secondary forest, including Macaranga denticulate forest (Md), Millettia laptobotrya forest (Ml), Musa acuminata forest (Ma) and Dendrocalamus membranaceae forest (Dm) in Xishuangbanna showed that Acari, Hymenoptera (ant) and Collembola were the dominant groups of soil arthropods on all study sites, but their proportions differed in different forests. For the common and rare groups of soil arthropods, there were some differences in four forests. We found fewer taxonomic groups (number of order), individuals and DG diversity index of soil arthropod communities in Dm secondary forest, as compared to another three forest types. There was no significant difference in number of groups and individuals and DG diversity index of soil arthropods among Md, Ml and Ma. The taxonomic diversity and population of soil arthropods were higher in top soil (0 5 cm) than in deeper layers. The research indicated that the dynamics of soil arthropods was correlated with the fluctuation of temperature and rainfall in this area. In general, the groups, populations and DG diversity index of soil arthropod communities were higher in dry season or early rainy season than in middle rainy season. There were some differences in the seasonal variation of soil arthropod communities in different type forests, which was primarily related to precipitation, temperature, litter quantity and quality, and biology of different soil arthropods. Apparently, food, habitation and disturb also have effects on the seasonal variation of soil arthropod communities in different forests. PMID- 15362622 TI - [Food nourishment utilization efficiency of different month aged sheep]. AB - Studies on the food nourishment utilization efficiency of different month aged sheep showed that under same food nourishment condition, the ingested nourishment, weight increase, and nourishment depletion per unit weight increase of sheep were increased with the ingestion of dry matter, and the protein content of 34 months old sheep was significantly higher than that of 22 months old sheep (P < 0.01), and the latter was significantly higher than that of 10 months old sheep (P < 0.01). There was a significant difference of utilization efficiency of the ingested nourishment between 34 months old and 22 months old sheep (P < 0.05) and between 10 months old and 22 months old sheep (P < 0.01). The weight increase was not significantly different (P > 0.05) between 34 months old and 22 months old sheep and between 10 months old and 22 months old sheep, but it was significantly different between 34 months old and 10 months old sheep (P < 0.05). The feed conversion rate among them was not different. All the results showed that 10-22 months old sheep had high production efficiency, and could produce high quality mutton, and hence, this section of age was the suitable age of sheep to be butchered. PMID- 15362623 TI - [Effect of fertilization on soil enzymes and microbes in Larix gemlinii plantations]. AB - One year-old second generation stand and 14 year- and 34 year-old first generation stands in larch plantations were studied at the Maoershan Mt. Experimental Forest Farm. The forestland of these three stands was treated with various fertilizations. The results showed that fertilization could promote or restrain soil enzyme activity and microbial amount with different degrees, and its effect on soil physiological activity was more apparent. The effect of the same fertilization treatments on soil enzyme activity and microbial amount was different at different development stages of the stands. For one year-old stand, the best fertilization scheme was treatment 9, its soil enzyme activities (catalase, proteinase, polyphenoloxidase, urease, and saccharase), total microbial amount, and amount of bacteria, actinomyces and fungus being increased by 413.49%, 22.10%, 20.56%, 220.00%, 49.46%, 238.88%, 247.24%, 106.70%, and 366.67%, respectively, as compared with the control. For 34 year-old stand, the best fertilization scheme was treatment 5, its soil enzyme activities (catalase, proteinase, polyphenoloxidase, urease, and saccharase), total microbial amount, and amount of bacteria and fungus was increased by 30.44%, 16.91%, 0.22%, 43.06%, 124.18%, 119.92%, 87.66%, and 17.57%, respectively in rhizosphere soil, and by 24.55%, 77.01%, 168.62%, 251.85%, 183.33%, 250.0%, 38.24% and 128.57%, respectively in non-rhizosphere soil. For 14 year-old young stand, it needed a proper amount of nitrogen fertilizer and organic mineral fertilizers, and the better fertilization schemes were treatments 2 and 9. The soil enzyme activities (catalase, proteinase, and urease) in treatment 2 was increased by 44.39%, 94.83%, and 4.62%, respectively in rhizosphere soil, and by 13.98%, 10.70% and 129.76%, respectively in non-rhizosphere soil. Total soil microbial amount and the amount of bacteria and fungus in treatment 9 was increased by 176.49%, 266.63%, and 198.04%, respectively in rhizosphere soil, and by 275.56%, 66.67% and 143.75%, respectively in non-rhizosphere soil. PMID- 15362624 TI - [Variation of microflora and enzyme activity in continuous cropping cucumber soil in solar greenhouse]. AB - Variation of microflora and enzyme activity in solar greenhouse soil continuous cropping for 1, 3, 5, 7, 9 years was studied, in addition to the relationship between soil properities and microflora and enzyme activity. The results showed that number of bacteria, actinomyce as well as total microflora showed a trend with a saddle-shaped curve, while the number of fungi appeared an liner increase. Continuous cropping soil microflora shifted from bacteria type to fungi type significantly, of which Ammoniation bacterium and Fusarium oxysporum were main physiology groups. Path analysis results showed that microelements (Mn, Cu, Fe), organic matter, available N and bulk density are main restricted factors of soil microflora and enzyme activity in solar greenhouse. PMID- 15362625 TI - [Effect of sodium phytate on alkaline phosphatase (ALP) activity in intraradical hyphae of AM fungi and development of its extraradical hyphae]. AB - In this paper, a pot experiment with three compartments was installed to study the effect of Na-phytate on the development and metabolic activity of AM fungi. Maize was selected as host plant and inoculated with arbuscular mycorrhizal fungus (Glomus intraradices), and different amounts of Na-phytate were applied into hyphal compartments. The results showed that the absorbed P in the shoots and roots of maize inoculated with AM fungi was 1-2 times higher than that of non inoculated maize. The ALP activity in intraradical hyphae of AM fungi was significantly decreased, and the total hyphal density of AM fungi increased with applied Na-phytate. Observations suggested that the ALP activity in intraradical hyphae of AM fungi and the development of its extraradical hyphae could be regulated by Na-phytate. Additionally, AM fungi could enhance the plant's P absorption and utilization from native soil P and additive Na-phytate. PMID- 15362626 TI - [Response of soil hydrolase and oxidoreductase activities to free-air carbon dioxide enrichment (FACE) under rice-wheat rotation]. AB - This paper studied the response of soil urease, phosphatase, arylsulphatase and dehydrogenase to 200 micromol x mol(-1) CO2 elevation under rice-wheat rotation. The results showed that under CO2 elevation, the urease activity in 0-10 cm soil layer was decreased at the early growth stages of wheat but increased at its booting stage; the activity increased at the early growth stages of rice but decreased at its ripening stage. Phosphatase activity was increased during the whole growth period of wheat; the activity increased at the tillering stage of rice but decreased at its later growth stages. Arylsulphatase activity was decreased at the over-wintering and booting stages of wheat but increased at its tillering and ripening stages. Dehydrogenase activity was decreased at the early growth stages of wheat and rice, but increased at their late growth stages. PMID- 15362627 TI - [Response of soil saccharidase activities to free-air carbon dioxide enrichment (FACE) under rice-wheat rotation]. AB - This paper studied the effect of 200 micromol x mol(-1) CO2 elevation on soil saccharidase activities and soil nutrient contents under rice-wheat rotation. The results showed that under both wheat and rice planting, CO2 elevation increased soil invertase activity. The elevated CO2 significantly increased soil xylanase activity at the jointing, heading and ripening stages of wheat and at the heading and ripening stages of rice, and slightly decreased soil cellulase activity. Correlation analysis showed that there was a significantly linear positive relationship between soil alkali-hydrolyzed nitrogen and soil invertase activity. PMID- 15362628 TI - [Vertical fractionation of dissolved organic matter in soil profile under water dry cultivation]. AB - Dissolved organic matter (DOM) is the most active component of organic matters, and plays an important role in the translocation and transformation of pollutants and nutrients in terrestrial ecosystem. In this paper, the vertical fractionation of DOM in a yellow brown soil under water-dry cultivation was investigated by field experiment and physico-chemical analyses. The results showed that in treatments CK, chemical fertilizer and sewage sludge, the DOM concentration decreased from 145.8, 117.7 and 114.8 mg x kg(-1) to 21.57, 23.23 and 13.78 mg x kg(-1) with soil depth, respectively. In control and fertilizer-treated soils, the hydrophilic fraction of DOM was increased respectively from 19.01% and 18.47% to 34.97% and 44.37% with soil depth, and the biodegradable rate of DOM also increased with soil depth. In sludge-treated soil, a reverse tendency was found for the concentration of hydrophilic DOM and its biodegradable rate. Hydrophilic fraction decreased from 36.96% to 17.07%, the increase of hydrophobic fraction was from 63.04% to 82.93%, and the biodegradable rate decreased from 53.19% to 30.75% with soil depth. The biodegradable rate of DOM was positively correlated to the hydrophilic fraction of DOM, and UV spectra analysis also suggested this. PMID- 15362629 TI - [Bacteria ecology in planting-culturing system]. AB - Planting-culturing system in inter-tidal zone is a new type eco-culturing model. The survey on bacteria biomass and water quality in the designed planting culturing system in inter-tidal zone showed that the mangrove planted in the system improved water quality and made water quality to II-III type, better than the IV and V type in the control pond. Designed ponds made heterotrophic bacteria, vibrio, phosphorus bacteria and enzyme-producing bacteria populations 1 2 order lower than the control pond without mongrove planting. Correlation analyses with CORREL software showed that the biomass of these bacteria was positively related with the nitrogen and phosphorus contents in water of the system, and the correlation coefficient for heterogeneous bacteria and vibrio was up to 0.9205. Heterotrophic bacteria and vibrio could be used as the water quality monitoring organisms. PMID- 15362630 TI - [Effect of temperature on Macrosiphoniella sanborni development]. AB - The development of Macrosiphoniella sanborni was studied at 15-35 degrees C and 80% RH, and the data obtained were used to determine the rate and threshold of its development and the sum of effective temperature. The optimal temperature for M. sanborni development ranged from 22 degrees C to 25 degrees C. After fitting the data to Wang Rusong's model, the development rate V(t) increased with rising temperature. The 2nd and 3rd instar nymphs were found more tolerable to extreme temperature and had higher optimal temperature than 1st and 4th instar nymphs. The temperature tolerance value delta was 6.70 for the 2nd and 3rd instars, and 4.70 and 4.50 for the 1st and 4th instars, respectively. The optimal temperature for the 2nd and 3rd instar was 21.76 degrees C, and the highest development temperature was 40.97 degrees C, while for the 1st and 4th instar, the optimal temperature was 23.76 degrees C and 22.49 degrees C, and the highest development temperature was 39.97 degrees C and 40.56 degrees C, respectively. Calculations with direct optimum estimation method showed that the threshold of development decreased with the aging of nymphs, while the effective accumulative temperature increased. For the 1st, 2nd, 3rd and 4th instar, the threshold of development was 6.93, 5.02, 4.58 and 4.46 degrees C, and the effective accumulative temperature was 26.88, 33.41, 33.63 and 48.49 degrees C x d(-1), respectively. PMID- 15362631 TI - [Spatial structure of populations Myzus persicae and its predatory natural enemy Erigonidium graminicola]. AB - In this paper, the spatial construction models of populations M. persicae and its predatory natural enemy E. graminicola during different periods were simulated by geostatistics, and their spatial relationships were analyzed. The spatial structure of M. persicae population was described by spherical model, showing an aggregated spatial arrangement. Its spatial dependence was 2.0252-4.1495 m, heterogeneity degree was 10,281.36-300,216.30, and sample variance was 12,176.81 303,433.70. The spatial structure of E . graminicola population was also simulated by spherical model, showing an aggregated spatial arrangement. Its spatial dependence was 3.7328-4.8983 m, heterogeneity degree was 1.4482-4.4134, and sample variance was 1.6941-5.8167. The results and methods could be applied to monitor the temporal and spatial dynamics of target insect pest population in tobacco field, and provide scientific basis for ecological control. PMID- 15362632 TI - [Geostatistic analysis on spatial patterns of Aphis gossypii and Erigonidium graminicola]. AB - The spatial structure and spatial correlativity of Aphis gossypii and Erigonidium graminicola populations during different periods were investigated by geostatistic methods. The results showed that the semivariogram of A. gossypii appeared as spherical type, indicating its spatial pattern was an aggregated type with a range of 7.33-12.40 cm, while that of E. graminicola was also fitted into spherical model, indicating an aggregated spatial arrangement with a range of 6.47-20.88 cm. The amount and spatial distribution of E. graminicola were closely related to those of A. gossypii, suggesting that E. graminicola was the dominant natural enemy influencing A. gossypii population. PMID- 15362633 TI - [Emergy evaluation of power plant eco-industrial park]. AB - In this paper, new emergy indices for the material circulation utilization and energy cascade utilization were presented to evaluate the emergy of power plant eco-industrial park. The common contribution of the members in power plant eco industrial park should be accounted for emergy yield ratio (PEYR) and emergy investment ratio (PEIR), namely, PEYR = sigma(i = 1)6 (Fi + Ri + Ni)/sigma(i = 1)6 Fi and PEIR = sigma(i = 1)6 Fi/sigma(i = 1)6 (Ni + Ri). Saved renewable and nonrenewable resources and purchased resources and decreased environmental load should be accounted for environmental loading ratio (PELR) and index of sustainability (PESI), namely, PELR = sigma(i = 1)6 Fi/F' and PESI = PEYR/PELR. Case analysis on the Shuozhou power plant eco-industrial park showed that new emergy indices were practical in evaluating the power plant eco-industrial park. PMID- 15362634 TI - [Concept and methodology on up-scaling issue of small watershed prototypes for reconstructing ecological economies in Loess Plateau]. AB - The general ecological situation of Loess Plateau is kept deteriorating, despite great efforts have been made and some achievements were gained after 50 years' endeavor to control soil erosion and renovate ecological landscape. Investigations on the past 20 years results from 11 experimental sites showed that the extension of prototypes and models derived from these sites, which were all based on one scale of small watershed, were unsatisfactory. The possible reasons for such a situation could be attributed to the too coarse scale of map, on which, the regional planning were set up and the locations of these experiment sites were decided, the too fine scale of the small watershed which were usually 3-10 km2 in size, the shortage of studies on various scales particularly on the meso-scale, and the lack of methods of scaling on up-scaling. Based on the analysis of restraints in management of small watershed, the specific concept and relevant proposed methods of resemblance-scaling and multi-scale complementarity were put forward and tested. PMID- 15362635 TI - [Sensitivity analysis in ecological modeling]. AB - Sensitivity analysis is used to qualitatively or quantitatively apportion the variation of model output to different source of variation. It is a very useful tool in model parameterization and calibration, and has important ecological significance by identifying the governing factors for a certain ecological process simulated. There are two schools of sensitivity analysis, local sensitivity analysis and global sensitivity analysis. The former examines the local response of the output(s) by varying input parameters one at a time, holding other parameters to a central value; and the latter examines the global response (averaged over the variation of all the parameters) of model output(s) by exploring a finite (or even an infinite) region. Since it is very easy to conduct local sensitivity analysis, it is very popular in ecological models. However, local sensitivity analysis is not computationally effective, because it can only get the sensitivity of a single parameter at a time. It can not take into consideration the effect of interaction of different parameters. Additionally, the value of other parameters will affect the sensitivity of the parameter specified. In view of this, global sensitivity analysis is increasingly preferred to local sensitivity in recent years. However, for most of the ecological modeling study published in Chinese, only local sensitivity analysis is conducted. To provide a toolbox of alternative sensitivity analysis algorithms for ecological model development in future study, we reviewed the main methods of both local sensitivity analysis and global sensitivity analysis, including one at a time method, multivariate regression, Morris' method, Sobol's method, Fourier Amplitude Sensitivity Analysis, and Extended Fourier Amplitude Sensitivity Analysis. Based on the state-of-the-art research on sensitivity analysis, the sensitivity of the interaction between two or more than two model parameters, the sensitivity of common model parameters in a set of models, and the sensitivity analysis in spatially explicit landscape model simulation were identified as the key areas and difficulties of future study on sensitivity analysis in ecological modeling. PMID- 15362636 TI - [Response of fine roots to soil nutrient spatial heterogeneity]. AB - The spatial heterogeneity is the complexity and variation of systems or their attributes, and the heterogeneity of soil nutrients is ubiquitous in all natural ecosystems. The scale of spatial heterogeneity varies considerably among different ecosystems, from tens of centimeters to hundred meters. Some of the scales can be detected by individual plant. Because the growth of individual plants can be strongly influenced by soil heterogeneity, it follows that the inter-specific competition should also be affected. During the long process of evolution, plants developed various plastic responses with their root system, including morphological, physiological and mycorrhizal plasticity, to maximize the nutrient acquisition from heterogeneous soil resources. Morphological plasticity, an adjustment in root system spatial allocation and architecture in response to spatial heterogeneous distribution of available soil resources, has been most intensively studied, and root proliferation in nutrient rich patches has been certified for many species. The species that do respond may have an increased rate of nutrient uptake, leading to a competitive advantage. Scale and precision are two important features employed in describing the size and foraging behavior of root system. It was hypothesized that scale and precision is negatively related, i. e., the species with high scale of root system tend to be a less precise forager. The outcomes of different research work have been diverse, far from reaching a consensus. Species with high scale are not necessarily less precise in fine root allocation, and vice versa. The proliferation of fine root in enriched micro-sites is species dependent, and also affected by other factors, such as patch attributes (size and nutrients concentration), nutrients, and overall soil fertility. Beside root proliferation in nutrient enriched patches, plants can also adapt themselves to the heterogeneous soil environment by altering other root characteristics such as fine root diameter, branch angle, length, and spatial architecture of root system. Physiological and mycorrhizal plasticity can add some influence on the morphological plasticity to some extent, but they are less studied. Roots located in different patches can quickly regulate their nutrient uptake kinetics within different nutrient patches, and increase overall nutrient uptake. Physiological response may, to certain extent, reduce morphological response, and is meaningful for plant growth on soils with frequently changing spatial and temporal heterogeneity. Mycorrhizal plasticity has been least studied so far. Some researches revealed that mycorrhiza, rather than fine root, proliferated in enriched patches. But, it is not the case with other studies. The proliferation of mycorrhiza within enriched patches is more profitable in term of carbon invest. The effect of fine root proliferation on nutrient uptake is complex, depending on ion mobility and whether or not neighboring plant exists. The influence of root plasticity on the growth of plants is species specific. Some species (sensitive species) gain growth benefit, while others don't. The ability of an individual plant to response to heterogeneous resources has significant effect on its competitive ability and its fate within the community, and eventually shapes the composition and structure of the community. PMID- 15362637 TI - [Formation of iron plaque on root surface and its effect on plant nutrition and ecological environment]. AB - This paper illustrated the conditions of iron plaque formation on root surface, the morphology, mineralogical composition and deposition site of the plaque, and its function as a Fe reservoir in supplying Fe nutrient and affecting the uptake of other nutrients such as phosphorus and zinc. The environmental and ecological role of the plaque in inhibiting the uptake and translocation of heavy metals such as Cu, Ni, Cd and As through absorption or co-precipitation was called external tolerance mechanism, while its competition with heavy metals for metabolically sensitive sites in plants was called internal tolerance mechanisms. These two mechanisms help plant survive in high acidic and low carbon environment. PMID- 15362638 TI - [Research advance in application of heat technique in studying stem sap flow]. AB - This paper reviewed the principles of heat technique and its application in studying stem sap flow. Heat technique combined with determinations of tree physiological items such as whole-tree hydraulic conductance, stomatal conductance, water potential, and stem water storage can make us deeply analyze the regulation mechanism of tree transpiration, and approach the effect of environmental conditions on stem sap flow and its response. In addition, this technique can be also used for a long-term measurement of the hydrological characters of zonal forest stands, which will give a powerful technical support in properly assessing the hydrological effect of forest. PMID- 15362639 TI - [Research advance in cyclic hydroxamic acids, main allelochemicals of Zea mays]. AB - The research advance in cyclic hydroxamic acids was reviewed in this paper. Cyclic hydroxamic acids are the important natural products of cereal crops. They and their respective derivatives are the constitutive compounds of a wide variety of gramineous plants and few dicot plants. They have structural diversity and different natural occurrences. Because of their phytotoxic properties, cyclic hydroxamic acids show a great variety of biological activities. They are the defensive agents against plant diseases, pests, nematodes and other plants. The distribution of cyclic hydroxamic acids in Zea mays and their variation in relation to the age were focused on in the paper. In Zea mays, there are structural diversity of cyclic hydroxamic acids and related benzoxazolinones. DIMBOA (1,4-benzoxazin-3(4H)-ones) is the most abundant derivative in Zea mays. The content of cyclic hydroxamic acids is strongly cultivar-dependent in Zea mays. Hydroxamic acids are not present in seeds. After germination, the level of DIMBOA increases, and the maximum level occurs in young seedlings a few days after germination. DIMBOA exists in all parts of plants, and its concentration is generally higher in shoots than in roots. In all stages, the young leaves of Zea mays have relatively high content of DIMBOA. The concentrations of these hydroxamic acids are highly dependent on environmental growth conditions. Under UV-light and water deficiencies, the levels of hydroxamic acids in plant increase rapidly. Cyclic hydroxamic acids exuded by Zea mays root can be quantitatively analyzed by HPLC. Supplying iron can significantly increase the exudation of DIMBOA from Zea mays root. PMID- 15362640 TI - [Soil dissolved organic matter and its effect on chemical and biological behaviors of soil heavy metals]. AB - Dissolved organic matter (DOM) is an important and active component in both terrestrial and aquatic ecosystems, while soil DOM plays an important role in the pedogenesis, mineral weathering, and translocation of pollutants in soils. It has become a hotspot in environment science, soil science and ecology. Soil DOM has important effects on the chemical and biological behaviors of heavy metals in soil-plant system, but its mechanism is not identified. The study on soil DOM in China was still in its initial stages, and most studies only reported that the activity of heavy metals in soils was affected by the DOM produced from organic fertilizer. This review is to summarize the recent literatures regarding the formation of soil DOM and its affecting factors, and the effect of soil DOM on the chemical and biological behaviors of heavy metals in soils. The main affecting mechanisms of soil DOM on behaviors of soil heavy metals were summarized as complexing with heavy metals, competing with metals for adsorption sites, and affecting soil pH value. PMID- 15362641 TI - [Research advance in characteristics of molybdenum, boron and selenium in red soils of South China and effects of their reasonable application on forage grass growth]. AB - Reasonable fertilization and fertility improvement of red soils are the important measures to develop the forage industry in hilly regions of South China. This paper summarized the existing form, chemical feature, availability and content of molybdenum, boron and selenium in red soils of South China. The lack of these three microelements in red soils was in common, and their reasonable application could significantly improve the growth of forage grass and its yield. The authors discussed the effects of molybdenum, boron and selenium application on the uptake and accumulation of available nutrients by forage grass and relevant physiological functions, described the molybdenum, boron and selenium deficiency symptoms of forage grass, their diagnostic methods and correcting measures, and put forward some suggestions for deepening the study. PMID- 15362642 TI - [Mechanism of stomatal regulation by root sourced signaling and its agricultural signficance]. AB - Under soil drought condition, root sourced signal abcisic acid (ABA) plays an important role in the long distance signaling process, and can be a measurement of soil water availability. ABA is also an effective stomatal closing agent, and acts to reduce transpiration and canopy water loss. This paper briefly introduced the physiological mechanism and theoretical model about the stomatal regulation by root sourced signaling, and indicated that the combination of this model with root water absorption model and stomatal conductance model could be more effective in depicting the response of plant to soil drying and atmospheric drought. In addition, some effective irrigation approaches, such as regulated deficit irrigation (RDI), partial root-zone drying (PRD) and controlled alternative irrigation (CAI) were profited from the mechanism of plant water use regulation by the root sourced signaling. These irrigation measures favored to reasonably distribute available soil water in root-zone. Root signaling system also played important role in regulating root growth and its development, retarding shoot growth to adjusting root shoot ratio, and optimizing assimilation allocation to favor to improve reproductive development. These processes hold substantial promise for enhancing crop water use efficiency. PMID- 15362643 TI - [Antibiotics production by bacterial agents and its role in biological control]. AB - Using bacteria to control plant diseases is one of the main strategies in plant protection, and its mechanism is commonly thought to be the production of antibiotics by bacteria. The produced antibiotics not only have structural diversity, but also have broad-spectrum activity against many pathogens. Experimental results showed that one kind of antibiotics could be produced by several bacterial strains, and one bacterial strain could also produced more than one kind of antibiotics. Recent development in molecular and bio-analytical techniques greatly promoted the research of bacterial bio-control, and the colonization survey of introducing bacterial strain to the rhizosphere and spermosphere of host plants. Besides bacterial genotypes, several biotic and abiotic factors including whether the pathogens existing or not, temperature, pH, carbon and nitrogen sources are also identified to affect the antibiotics production by bacteria. The authors illustrated some of the impeding problems in bacterial bio-control agents' application, and suggested that in the future research, more attention should be paid on developing mixed bio-control agent to avoid the anti-antibiotic activity of pathogens. PMID- 15362644 TI - [Community characteristics in Hongchiba area of Wuxi County during ecological restoration after controlled burning]. AB - This paper studied the species composition and the structure and species diversity of communities in Hongchiba area of Wuxi County after controlled burning and aerial-sowing afforestation. The results showed that after burning and afforestation, pure Pinus armandii forest was not developed, but various mixed needle board-leaved forests which comprised P. armandii and native board leaved trees grew. Various native species contributed to the communities with great species diversity. Shannon-Wiener index was 2.305-3.145, ecological dominance was 0.063-0.151, and evenness was 0.600-0.749. The natural regeneration of P. armandii population was very poor, and there was a trend that P. armandii would be gradually replaced by other native board-leaved trees. It was demonstrated that P. armandii should not be taken as a unique afforest tree, and controlled burning was not a favorite ecological measure in subtropical area. Native trees should play more important roles in the process of ecological restoration. PMID- 15362645 TI - [Life table and spectral analysis of endangered plant Taxus chinensis var. mairei population]. AB - Based on the investigation in Longxi Mountain National Nature Reserve and the theory of survival analysis, a static life table of Taxus chinensis var. mairei population was worked out, the curves of its survival rate, mortality rate and killing power were drawn, and the population dynamics was analyzed by spectral analysis. The results showed that the survival curve of the population appeared to be a type of Deevey-III, and the high mortality of seeding was one of the important reasons which caused Taxus chinensis var. mairei to be endangered. The spectral analysis of the population showed that there was a marked periodic regularity in the process of natural regeneration of Taxus chinensis var. mairei. PMID- 15362646 TI - [Child endangerment by maternal alcohol, nicotine and drug abuse]. PMID- 15362647 TI - [General nursing practice in treating neonatal jaundice]. PMID- 15362648 TI - [Pain observation and assessment in premature infants]. PMID- 15362649 TI - [Expectations of nursing care from the viewpoint of young patients]. PMID- 15362650 TI - [Psychological stress of parents of premature infants]. PMID- 15362651 TI - [Basic principals of terminal care by the physician. Revision by the Federal Physicians Office]. PMID- 15362652 TI - [Preventive examinations are freed from practice tax]. PMID- 15362653 TI - [Encouraging life--allowing dying]. PMID- 15362654 TI - [New chances for pediatric nursing care: model project in Halle by the Saale]. PMID- 15362655 TI - Impact of host genetics on HIV disease progression and treatment: new conflicts on an ancient battleground. PMID- 15362656 TI - Peripheral neuropathy in lentivirus infection: evidence of inflammation and axonal injury. AB - OBJECTIVE: As distal sensory polyneuropathy (DSP) is a major neurological complication of HIV-1 infection, we investigated the extent of peripheral nervous system disease in animals infected with the lentivirus, feline immunodeficiency virus (FIV), because it causes neurological disease and immunosuppression in cats similar to HIV-1 in humans. METHODS: After infection with a neurovirulent FIV molecular clone, neurobehavioral testing, nerve morphology, viral detection and load measurements were performed. RESULTS: Neurobehavioral studies showed delayed withdrawal in response to a noxious stimulus among FIV-infected animals compared with sham-infected controls (P < 0.05). Dorsal root ganglia and sciatic nerves from FIV-infected ammals showed activated macrophages that were increased in number and size compared with controls. In addition, TNF-alpha messenger RNA was detectable in most nerves and spinal cords from the FIV-infected group, but was infrequently detected in controls. Viral RNA copy numbers in plasma and sciatic nerves were detectable in all FIV-infected animals at high levels. Studies of sural nerves identified myelinated fiber atrophy in 12-week FIV-infected animals compared with age-matched control animals, which was accompanied by reduced myelin sheath thickness (P < 0.05). The footpads of FIV-infected animals displayed reduced intraepidermal fiber density compared with control animals (P < 0.01). CONCLUSION: FIV infection results in the rapid onset of peripheral neuropathy, defined by axonal injury and macrophage activation, together with abundant virus within the nerve, indicating that it may serve as a model of HIV related DSP. PMID- 15362657 TI - Enhanced HIV-specific immune responses in chronically HIV-infected patients receiving didanosine plus hydroxyurea. AB - BACKGROUND: The role of hydroxyurea (HU) in the treatment of HIV infection remains controversial. HU potentiates didanosine (ddI) antiviral activity and might exert immunomodulatory effects. PATIENTS AND METHODS: Immunologic parameters were examined in HIV-infected patients enrolled in a simplification trial in which ddI-HU was provided to subjects who had been on complete virus suppression under highly active antiretroviral therapy (HAART) for longer than 6 months. A total of 84 of these patients showed plasma viraemia repeatedly below 5000 HIV-RNA copies/ml, and were the main study population. A group of 22 patients who continued on HAART and another of 22 drug-naive HIV-positive individuals were taken as controls. RESULTS: At 12 months, the levels of naive and memory T-cell subsets were similar in patients on ddI-HU and under HAART, whereas immune activation tended to be lower in the former group. The frequency of HIV-specific CD8+ T cells (CTL) directed against 125 peptides derived from Gag, Pol, Env, Nef and HIV regulatory proteins was similar among patients on ddI HU and untreated controls, and significantly higher than in patients under HAART. This higher CTL response in patients on ddI-HU was seen even when considering only subjects with undetectable viral load. HIV-specific CD4+ T-cell responses were absent in almost all patients on HAART, whereas they were present in up to 19% of patients on ddI-HU. CONCLUSION: Treatment with ddI-HU provides higher levels of HIV-specific CD8+ and CD4+ T-cell responses than standard triple drug regimens. Thus, hydroxyurea might exert a beneficial immunomodulatory effect in HIV infection. PMID- 15362658 TI - Anti-HHV-8/KSHV antibodies in infected individuals inhibit infection in vitro. AB - OBJECTIVE: To detect human herpesvirus (HHV)-8/Kaposi's sarcoma-associated herpesvirus (KSHV) neutralizing antibodies (nAb). DESIGN: Antibodies capable of inhibiting HHV-8 infection were measured by infecting transformed dermal microvascular endothelial cells (tDMVEC) with HHV-8 that had been pre-incubated with serum from HHV-8-seropositive or -seronegative subjects. The level of infection was quantified 48 h later. METHODS: HHV-8 was prepared from JSC-1 primary effusion lymphoma cells; the titre of enveloped virions was determined by electron microscopy. Virus was incubated with serum samples for 60 min before inoculating tDMVEC. The level of infection was quantified by indirect immunofluorescence assay, staining for HHV-8 latency-associated nuclear antigen (LANA)-1. Inhibition of infection was determined by comparing the level of infection obtained with HHV-8-seropositive subject serum with the level obtained by incubation with seronegative serum. RESULTS: Up to 61% of cells were infected with HHV-8 in the absence of human serum; this level was not affected by pre incubating the virus with HHV-8-seronegative serum. At dilutions of 1:10 and 1:50, HHV-8-seropositive sera significantly inhibited infection compared to seronegative controls (P = 0.036 for both serum dilutions, Mann-Whitney). The endpoint of inhibition was 1:100, when the serum of one of five subjects inhibited virus infection. At 1:500 dilution, there was no difference in the level of infection after virus incubation with seropositive or seronegative sera (P = 0.578). Depletion of antibody from serum with protein A reversed the inhibitory effect, confirming it was antibody-mediated. CONCLUSIONS: This study is the first to identify HHV-8 antibodies in infected subjects that reduce in vitro infectivity of the virus. PMID- 15362659 TI - Inhibition of herpesvirus-induced HIV-1 replication by cyclopentenone prostaglandins: role of IkappaB kinase (IKK). AB - OBJECTIVES: Herpes simplex virus (HSV) infections have been associated with reactivation of HIV-1 replication and increases of HIV-1-load in plasma of co infected individuals. The present authors have previously reported that in epithelial cells HSV-1 induces the IkappaB-kinase (IKK) causing persistent activation of NF-kappaB, a critical regulator of HIV-1 replication. The present study was performed to investigate whether HSV-1-infection could induce IKK mediated NF-kappaB activation and enhance HIV-1 expression in human T cells, and to analyze the effect of the IKK-inhibitor prostaglandin A1 (PGA1) and other prostanoids on the NF-kappaB-mediated HSV-HIV interaction. DESIGN AND METHODS: Induction of IKK and NF-kappaB activity was determined in lymphoblastoid Jurkat cells and HIV-1 chronically-infected H9 and ACH-2 cells by kinase assay and electrophoretic mobility shift assay, respectively. The effect of HSV-1 and different prostanoids on HIV-1 expression and replication was determined in Jurkat cells transfected with HIV-1-LTR-driven reporter genes, and in H9 and ACH 2 cells by p24-antigen level evaluation. The role of NF-kappaB in HSV-1-induced HIV-1 expression was investigated by using the IkappaBalpha dominant-negative IkappaBalpha-AA in co-transfection experiments. RESULTS: In human T lymphoblastoid cells HSV-1 potently induces IKK activity, causing a persistent induction of NF-kappaB. HSV-1-induced IKK and NF-kappaB function results in transactivation of HIV-1-LTR-regulated genes and induction of HIV-1 replication in chronically-infected T cells. The cyclopentenone PGA1 inhibits HSV-1-induced IKK and NF-kappaB activities, blocking HIV-1-LTR-driven expression and preventing HSV-1-induced HIV-1 replication in co-infected cells. CONCLUSIONS: The results indicate that IKK is a key factor in triggering HSV-1-induced HIV-1 transcription in chronically-infected cells and identify cyclopentenone prostanoids as potent inhibitors of HSV-1-induced HIV-1 reactivation. PMID- 15362660 TI - Maternal versus paternal inheritance of HLA class I alleles among HIV-infected children: consequences for clinical disease progression. AB - OBJECTIVE: When children acquire HIV infection from their mothers (with whom they share at least 50% of their HLA alleles), they acquire virus with a history of encounter with maternal HLA-mediated immune responses. We investigated whether maternal HLA selection pressures on the virus would adversely influence clinical outcomes of HIV-infected children. METHODS: We tested whether time to AIDS diagnosis or death, among a cohort of 59 HIV-infected children in New York City followed from birth for up to 12 years, was associated with maternally- or paternally-inherited child HLA class I alleles, and with HLA similarity between mother and child. RESULTS: HIV-infected children with an HLA allele usually associated with slow disease experienced a slower progression to AIDS or death only if the allele was paternally inherited. If the allele was present in the mother, no association was observed. Children who were homozygous or who shared both alleles with their mothers at more than one HLA class I locus were more likely to progress to AIDS or death than other children (relative hazard, 3.46; 95% confidence interval, 1.24-9.71). CONCLUSION: Genetic similarity between mother and child may compromise the child's capacity to control HIV replication when the virus is acquired from the mother. HLA-mediated selective pressures on the virus in a transmitting mother-infant pair may undermine future HLA-mediated viral control in the child. PMID- 15362661 TI - Atazanavir enhances saquinavir hard-gel concentrations in a ritonavir-boosted once-daily regimen. AB - OBJECTIVE: To determine the pharmacokinetics of saquinavir hard-gel capsules/ritonavir/atazanavir co-administered once daily at 1600/100/300 mg in HIV-infected individuals. METHODS: Eighteen patients receiving saquinavir/ritonavir switched to 1600/100 mg once daily a minimum of 3 days before the study. On study day 1, levels of saquinavir and ritonavir were determined over 24 h. Atazanavir (300 mg once daily) was then added to the regimen. On day 11, a pharmacokinetic analysis was performed. Atazanavir was discontinued on day 32. Drug concentrations were measured by high-pressure liquid chromatography-tandem mass spectrometry. Geometric mean ratios (GMR) and 95% confidence intervals (CI) were used to compare saquinavir and ritonavir pharmacokinetic parameters, with and without atazanavir. A safety analysis was performed at screening, days 1, 11, 32 and follow-up. RESULTS: After the addition of atazanavir, statistically significant increases in saquinavir trough plasma concentration (Ctrough GMR, 95% CI 2.12, 1.72-3.50), maximum plasma concentration (Cmax 1.42, 1.24-1.94), area under the plasma concentration-time curve from 0-24 h (AUC0-24 1.60, 1.35-2.43) and ritonavir Cmax (1.58, 1.32-2.08), AUC0-24 (1.41, 1.22-1.74) were observed. The pharmacokinetics of atazanavir compared with those obtained in patients receiving atazanavir/ritonavir without saquinavir. Four patients developed scleral icterus and two jaundice. Total and unconjugated bilirubin increased approximately fivefold during atazanavir therapy. CONCLUSION: The addition of atazanavir to saquinavir/ritonavir increased saquinavir Ctrough, Cmax and AUC0-24 by 112, 42 and 60%. Ritonavir Cmax and AUCo-24 increased by 34 and 41%. The regimen was well tolerated, with no significant change in laboratory parameters, except for the occurrence of hyperbilirubinemia. PMID- 15362662 TI - MIV-310 reduces HIV viral load in patients failing multiple antiretroviral therapy: results from a 4-week phase II study. AB - BACKGROUND: Drug resistance is an increasing problem in the treatment of HIV infection. MIV-310 (alovudine), a nucleoside reverse transcriptase inhibitor, potently inhibits the replication of highly mutated strains of HIV in vitro. We examined the efficacy of MIV-310 in highly pretreated patients. METHODS: In a phase II pilot study, 15 patients failing a current antiretroviral therapy with at least two thymidine-associated mutations (TAM) were given MIV-310 7.5 mg once daily for 4 weeks, in addition to their ongoing treatment. The primary endpoint was the plasma viral load reduction at week 4. RESULTS: At baseline, the median viral load was 3.93 log10 copies/ml and the median CD4 cell count was 360 cells/mm3. After 4 weeks of MIV-310 administration, the median decrease in viral load was -1.13 log10. Interestingly, the median reduction was only -0.57 log10 in the four patients on stavudine, contrasting with a median reduction of -1.88 log10 in the 11 patients not receiving concomitant stavudine. The viral load fell by a median of -1.60 log10 in patients with two to three TAM (n = 7), and by 1.88 log10 in patients with four or five TAM (n = 8). The viral load rebounded in all patients after MIV-310 cessation. No mutations were found in the reverse transcriptase coding region during MIV-310 treatment. MIV-310 was well tolerated, with no serious adverse events and no treatment withdrawals. CONCLUSION: MIV-310 7.5 mg/day efficiently reduced the HIV viral load in patients failing a multiple drug regimen. Further studies with different dosages and longer administration times are urgently needed. PMID- 15362663 TI - Virological, intracellular and plasma pharmacological parameters predicting response to lopinavir/ritonavir (KALEPHAR study). AB - OBJECTIVES: To assess the impact of HIV-1 protease mutations and intracellular and plasma lopinavir minimum concentrations (Cmin) on virological success or failure on lopinavir/ritonavir-containing highly active antiretroviral therapy (HAART). DESIGN: HIV-1-infected HAART-experienced patients included in an observational study, received lopinavir/ritonavir (400/100 mg twice a day) plus two to three nucleoside reverse transcriptase inhibitors (NRTI) or one NRTI plus one non-NRTI. A viral load less than 50 copies/ml at month 6 defined virological success. METHODS: Intracellular and plasma lopinavir concentrations were determined by high-pressure liquid chromatography with mass-spectrometry detection. Reverse transcriptase and protease genes were sequenced at baseline and the time of virological failure. RESULTS: When the 38 patients started the lopinavir/ritonavir-based regimen, baseline median (25-75th percentile) values were: CD4 cell count 218 cells/microl (133-477); plasma HIV-1-RNA load 5.3 log10 copies/ml (3.8-5.1); number of lopinavir mutations four per protease gene (two to six). Univariate analysis associated virological success or failure at month 6 (21/38 patients) with the number of baseline lopinavir mutations, intracellular and plasma lopinavir Cmin, and the genotype inhibitory quotient (GIQ) at months 1 and 6. Multivariate analysis showed that the number of baseline lopinavir mutations and intracellular and plasma lopinavir Cmin were independently associated with virological success or failure. We defined the most discriminating intracellular and plasma lopinavir Cmin efficacy thresholds (8 and 4 microg/ml, respectively) and GIQ thresholds (1 and 3, respectively). CONCLUSION: The monitoring of lopinavir/rironavir-based HAART efficacy should include the number of baseline lopinavir/ritonavir mutations, intracellular and plasma lopinavir Cmin and GIQ calculation. PMID- 15362664 TI - Primary HIV infection as source of HIV transmission within steady and casual partnerships among homosexual men. AB - OBJECTIVE: To assess the contribution of primary or acute HIV infection to the transmission of HIV among homosexual men in Amsterdam and to investigate how the initiation of treatment during primary HIV infection (PHI) can affect the incidence of HIV infection. METHODS: A mathematical model describing HIV transmission among homosexual men was developed. In the model, men are involved in both steady and casual partnerships. Infectivity is higher during PHI than during chronic HIV infection. Highly active antiretroviral therapy reduces infectivity and increases the time to the development of AIDS. Its effect is enhanced if treatment is initiated during PHI. HIV incidence and the fraction of transmission attributed to PHI were calculated for different levels of treatment efficacy. RESULTS: Primary infections account for 35% of HIV transmissions from casual partners and 6% of transmissions from steady partners. Among all new infections, only 11% occurs during PHI. Therefore, the effect of treatment during PHI on the incidence of HIV is limited. However, in a community with higher risky behaviour among casual partners, the fraction of transmission attributed to PHI increases to 25%. CONCLUSION: Primary infections play a more important role in transmission from casual partners than in transmission from steady partners. Therefore, in communities in which steady partners account for the majority of new infections and the epidemic is at an advanced phase, the contribution of PHI to the transmission of HIV is rather small and the effect of early treatment on the incidence of HIV is limited. PMID- 15362665 TI - Impact of HAART-related side effects on unsafe sexual behaviours in HIV-infected injecting drug users: 7-year follow up. AB - OBJECTIVE: To investigate the impact of non-lipodystrophy HAART-related side effects on unprotected sexual behaviours among HIV-infected drug users. DESIGNAND PARTICIPANTS: HAART-treated patients who reported having occasional partners during the follow-up period after HAART initiation were selected among patients of the MANIF 2000 cohort of HIV-infected drug users. METHODS: Visits where patients reported unsafe sexual behaviours with occasional partners were compared to visits where they reported safe sexual behaviours using a logistic model based on Generalized Estimating Equations. RESULTS: One-hundred and ninety-two HAART treated patients reported occasional sexual partners at least once during follow up, accounting for a total of 464 visits. Among these 192 patients, 134 (70%) declared at least once unsafe sexual behaviours with occasional partners. During follow-up, three or more HAART-related side effects were reported in 273 of the 464 visits. When comparing visits where patients reported unsafe sexual behaviours with occasional partners (n = 249) with those where they reported safe sexual behaviours (n = 215), experiencing three or more HAART-related side effects was significantly associated with unsafe sex after adjustment for cofactors such as injecting drug status, reporting more than two sexual partners and having sex more than once a week. CONCLUSIONS: Perceived side effects play a role in determining unsafe sexual behaviours. HIV prevention interventions must consider the negative impact of HAART-related side effects on sexual risk-taking behaviours. Drug maintenance programs contribute to sexual risk reduction among drug injecting patients. PMID- 15362666 TI - CCR5 promoter human haplogroups associated with HIV-1 disease progression in Thai injection drug users. AB - BACKGROUND: An evolutionary-based analysis of the CC chemokine receptor 5 gene (CCR5) promoter region has identified nine stable human haplogroups, within which certain haplogroups appear to influence HIV-1 disease progression differentially among Caucasians and African-Americans. OBJECTIVE: To assess the influence of CCR5 haplogroups on HIV-1 disease progression in a Thai population. DESIGN: Haplogroup analysis of HIV-1-seropositive injection drug users (IDU) participating in a prospective cohort study in Bangkok. All were documented seroconverters with a median follow-up time of 3.5 years (range, 0.2-7.0). METHODS: From a cohort of 130 IDU, 106 (81.5%) were genotyped for the CCR2b-64I, CCR5-delta32 and seven CCR5 promoter alleles constituting the CCR5 haplogroups. Survival curves and adjusted Cox proportional hazards models were used to assess the effect of haplogroups on the time from HIV-1 infection until CD4 count < 200 x 10(6) cells/l. RESULTS: The most common CCR5 haplogroups were HHC (61.8%), followed by HHE (15.6%) and HHF*2 (14.6%). HHE was associated with an accelerated CD4 count decline to < 200 x 10(6) cells/l (adjusted relative hazard, 1.88; 95% confidence interval, 1.05-3.36; P = 0.02). CONCLUSIONS: This is the first evidence that the CCR5 haplogroup E speeds the decline of the CD4 cell count and may lead to accelerated disease progression among HIV-infected Thais. These new observations highlight the need for additional studies involving populations in Asia. PMID- 15362667 TI - Decreased apoptosis of bone marrow progenitor cells in HIV-1-infected patients during highly active antiretroviral therapy. AB - Impaired haematopoiesis during HIV-1 infection may be caused by the overproduction of inflammatory cytokines by immune cells at the bone marrow level inducing Fas-mediated apoptosis of stem progenitors. In this study, we evaluated the effects of highly active antiretroviral therapy on apoptosis of CD34+ stem cells derived from the bone marrow of HIV-1-infected patients, and observed decreased Fas expression on progenitor cells, in parallel with the diminution of TNF-alpha levels and the amelioration of clonogenic parameters. PMID- 15362668 TI - IL-23 and IFN-gamma deficiency in immunodeficient HIV patients who achieved a long-term increase in CD4 T-cell counts on highly active antiretroviral therapy. AB - The pathogenesis of HIV infection and the susceptibility to opportunistic infections has been associated with poor type 1 cytokine production. In severely immunodeficient HIV patients who achieved increased CD4 T-cell counts on longterm highly active antiretroviral therapy, we observed reduced expression of IL-23p19 and IFN-gamma messenger RNA. Impaired IL-23-induced IFN-gamma production by memory T cells might thus contribute to opportunistic infections in a minority of patients with substantial CD4 T-cell recovery. PMID- 15362669 TI - Characterization of genotypic determinants in HR-1 and HR-2 gp41 domains in individuals with persistent HIV viraemia under T-20. AB - We analysed the genetic changes in gp41 in a population of 30 heavily pretreated HIV-1-infected patients failing on a T-20-containing salvage therapy. This study confirms the key role of the number of drugs presumed to be still effective associated with T-20 in the magnitude of the virological response. Our results suggest that only HR-1 sequencing is necessary to characterize resistance to T 20, and that HR-2 domain sequencing is probably not required. PMID- 15362670 TI - Maintenance therapy with cotrimoxazole for toxoplasmic encephalitis in the era of highly active antiretroviral therapy. AB - To reduce the number of daily pills for improving adherence to antiretrovirals, 17 protease inhibitor-treated patients receiving toxoplasmic encephalitis (TE) standard maintenance therapy were instead given cotrimoxazole 960 mg twice daily. After a median follow-up of 31 months, one relapsed after three months, TE relapse incidence = 2.1 cases per 100 patient-years (95% confidence interval, 0.05-11.3). This strategy could be useful for patients awaiting immune reconstitution which allows the interruption of TE maintenance therapy. PMID- 15362672 TI - Is the Vpr R77Q mutation associated with long-term non-progression of HIV infection? PMID- 15362673 TI - Gynecomastia and potent antiretroviral therapy. PMID- 15362674 TI - Emergence of Sjogren's syndrome in AIDS patients during highly active antiretroviral therapy. PMID- 15362675 TI - [A study on the expression of IL-8 and IL-3 in human nasal polyp and polyposis]. AB - OBJECTIVE: To study the expression of IL-8 and IL-3 in human nasal polyp and polyposis. METHOD: Using immunohistochemical methods detect the expressions of IL 8 and IL-3 in 52 cases of middle turbinate,nasal polyp and nasal polyposis,and calculating the positive area percentages in each group. All datas were analysed with SNK test, Kruskal-Wallis test and Mann-Whitney test. RESULT: There were very significant differences between either two groups about the expressions of IL-8. There were significant differences between either two groups about the expressions of IL-3. CONCLUSION: IL-8 is not only one of the causes in nasal polyp, but also plays an important role in the recurrence of nasal polyposis. The expression of IL-8 in polyp is different from that in polyposis, it indicate that polyp and polyposis have different mechanism. IL-3 is important for the accumulation of eosinophil and is a key factor in the mechanism of polyposis. PMID- 15362671 TI - Natural polymorphism of HIV-1 subtype G protease and cleavage sites. PMID- 15362676 TI - [The regulation of PKCalpha in eosinophil infiltration and proliferation in nasal polyps]. AB - OBJECTIVE: To explore the regulation of protein kinase C (PKC) isoform--PKCalpha in eosinophil (EOS) proliferation and infiltration in nasal polyp tissues. METHOD: With the methods of in situ hybridization staining and immunohistochemistry MGG staining, to check out the relationship between PKC and bcl-2/BaxmRNA and associated protein, especially PKC isoform--PKCalpha, PKCbeta1 , PKCbeta2, and PKCgamma did not express at all. RESULT: There were PKC expression in the eosinophils of 26 cases from nasal polyps, and the expression of PKC and Bcl-2 mRNA/their protein in EOS of nasal polyps showed remarkably positive relation (r1 = 0.0875, r2 = 0.0823, P < 0.01), but in PKC isoforms, PKCalpha expression was the strongest, but PKCbeta1 and PKCbeta2 expressed thinner and PKCgamma did not express at all. CONCLUSION: The reason of eosinophil proliferation and infiltration in nasal polyps is that PKC signal transduction pathway was activated, and leaded to inhibition of eosinophil apoptosis, and eosinophil survival was delayed, and eosinophil proliferated and infiltrated, and in PKC family, PKCalpha is main. PMID- 15362677 TI - [Role of ICAM-1 in eosinophilia and prognosis of nasal polyps]. AB - OBJECTIVE: To investigate the role of adhesion molecule ICAM-1 (Intercellular adhesion molecule-1) in eosinophilia and prognosis of nasal polyps. METHOD: Expression of ICAM-1 were detected and quantified in 35 cases of nasal polyps with fluorescence-immunohistochemistry analysis and its relationships to eosinophilia and prognosis of nasal polyps were evaluated. RESULT: The numbers of eosinophils in nasal polyps and control nasal mucosas were 28.4 +/- 10.7/mm2 and 9.9 +/- 8.6/mm2, and fluorescent densities of ICAM-1 were 115.9 +/- 82.4 and 28.2 +/- 17.1. The expression of ICAM-1 was associated with the heaviness of nasal polyps. CONCLUSION: ICAM-1 may play an important role in the formation and prognosis of nasal polyps. PMID- 15362678 TI - [Effects of dexamethasone on mucin gene expression in human nasal polyps]. AB - OBJECTIVE: To clarify whether glucocorticoid influences mucin 8 gene expression in human nasal polyps. METHOD: Thirty patients were randomly divided into two groups. The first group were given dexamethasone intravenously before surgery. We make sure the second group of patients didn't receive any kind of glucocorticoid therapy during thirty days before surgery. MUC8 MRNA expression in nasal polyps of each group was detected by reverse transcription polymerase chain reaction (RT PCR). RESULT: The MUC8 MRNA expression in the first group (0.71 +/- 0.34) was lower than that of the second group (1.12 +/- -0.26). CONCLUSION: DXM supress MUC8 MRNA expression in nasal polyps and this result provide another aspect for glucocorticoid's antiinflammatory mechanism. PMID- 15362679 TI - [Expression of nitric oxide synthase and its significance in nasal polyps]. AB - OBJECTIVE: To study the expression of nitric oxide synthase (NOS) and its activity in nasal polyps and its significance. METHOD: Septectomy and immunohistochemisty were used to examine NOS activity and identify the expression of inducible NOS (iNOS) and endotheial NOS (eNOS) in human nasal specimens from patients undergoing nasal spectrumtomy (n = 20) or nasal polypectomy (n = 30). RESULT: The activity of NOS was significantly higher in nasal polyps than in nasal mucosa (4.079 +/- 0.655) U/Pro vs (1.526 +/- 0.310) U/Pro (P<0.01). iNOS reaction product was located in epithelial cells, vascular endothelial cells, nasal glands and inflammation cells of nasal polyps. iNOS reaction product almost was not be founded in nasal mucosa. Amount of. iNOS positive cells in nasal polyps was higher than in nasal mucosa (P<0.01). eNOS reaction product were located in nasal polyps and nasal mucosa. There was no quantitative difference of eNOS between in nasal polyps and nasal mucosa. Amount of iNOS positive cells was significantly higher than of eNOS positive cells in nasal polyps (P<0.01). CONCLUSION: iNOS, mainly expressed in nasal polyps, locates in epithelial cells and vascular endothelial cells and nasal glands and inflammation cells in nasal polyps, which produce NO at high concentration that plays an important role in the formation of nasal polyps. PMID- 15362680 TI - [Approach to the maxillary sinus, the peterygopalatofossa for resecting maxillary nerve under the endoscopy]. AB - OBJECTIVE: This paper presents a method for treating maxillary neuralgia, the second division of trigeminal nerve, in the weak aged. METHOD: Thirty-nine cases with forty trigeminal neuralgia were treated from 1996 to 2001. All patients were treated through the maxillary sinus, the peterygopalatofossa for resecting maxillary nerve with endoscopic surgery. RESULT: The pain of all patients was relieved after operation and no recurrence of neuralgia was occurred in more than 2-years follow-up. CONCLUSION: This method had the advantages of avoiding to operate craniotomy, no complications, performing easily and the ideal treatments remained. PMID- 15362681 TI - [Analysis of clinical manifestations in nasal NK/T-cell lymphomas]. AB - OBJECTIVE: To investigate the clinical findings, histopathological features, treatment outcome as well as prognosis of nasal NK/T-cell lymphomas and to provide experiences for the diagnosis and treatment of the tumor. METHOD: Nine cases with nasal NK/T-cell lymphomas were treated from 1990-2000. Morphological and immunohistochemical study were performed on paraffin sections. Patients underwent radiation therapy (1 case), chemotherapy (2 cases), combination of radiation and chemotherapy (6 cases), respectively. RESULT: Clinically, NK/T-cell lymphomas had a male predominance and most presented as ulceration of lower turbinates and nasal septum. Histopathologically, NK/T-cell lymphomas were characterized by angiocentricity, angioinvasion and necrosis. The immunophenotyes of the NK/T-cell lymphomas were CD45RO+, CD43+, CD3epsilon+, CD56+. All patients with NK/ T-cell lymphoma died, and the duration of the disease from onset to death ranged from 3-34 months. CONCLUSION: Nasal NK/T-cell appear to have a male predominance, to originate more frequently in lower turbinates and nasal septum. They appears to carry a high malignancy and poor prognosis. NK/T-cell lymphomas pursue a aggressive course. PMID- 15362682 TI - [Evaluation of CT and MRI in diagnosis of intranasal encephalomeningocele]. AB - OBJECTIVE: To evaluate the values of CT and MRI in diagnosing intranasal encephalomeningocele. METHOD: The CT and MRI features of 7 cases confirmed by operation and pathology were retrospectively analyzed. RESULT: CT and MRI demonstrated that all cases were encephalomeningocele and 1 case was accompanied with hydrocephalus. MRI demonstrated that the herniation in nasal cavity were glial tissue and meninges, the signals are abnormal cerebrospinal fluid signal and it was attached to intracranial subarachnoid cavity. CONCLUSION: CT and MRI could diagnose encephalomeningocele correctly. CT could clearly manifest the size and location of the defect of cranial base bone, and the location of hernia. MRI was more advantageous than CT in displaying the contents in hernia, and the relationship between hernia and cranial base. Coronal and axial imaging planes with MR T1- and T2-weighted sequences are required to demonstrate intracerebral connection. PMID- 15362683 TI - [The expression of nm23-H1 in the investing papilloma and it's cancerated tissue of nasal cavity and parasinuses]. AB - OBJECTIVE: To investigate the role of the nm23-H1 protein in the development of investing papilloma and it's cancerated tissue in nasal cavity and parasinuses. METHOD: The expression of nm23-H1 protein was measured by immunohistochemistry in twenty-five cases with investing papilloma and twenty-one cases with cancerated investing papilloma in nasal cavity and parasinuses and fourteen cases with chronic sinusitis. RESULT: The expression of nm23-H1 in the investing papilloma was higher than that in cancerated investing papilloma tissue, the positive rate was 72% and 42% respectively in the two groups, P < 0.05. The expression of nm23 H1 in the investing papilloma was higher than in the chronic sinusitis, the positive rate of the latter was 14.3%, P < 0.05. CONCLUSION: The lower expression the nm23-H1 protein in the cancerated tissue of investing papilloma was related to the infiltration and canceration. PMID- 15362684 TI - [Ciliogenesis in human nasal epithelial cells cultured at the air-liquid interface]. AB - OBJECTIVE: In the present study, our aim was to develop a method to quantitative cell differentiation in cultures of human nasal epithelium (HNE) cells. METHOD: HNE cells were cultured on collagengel-coated membranes at an air-liquid interface (ALI) in hormone-and growth factor-supplemented medium. The percentage of the culture surface covered with ciliated cells was estimated using scanning electron microscope and image analysis. RESULT: In present study, if an ALI was not established and the cells were maintained in the submerged state, ciliated cell differentiation was pool, the average ciliated surface area was 0.3% when cultures were submerged for fourteen days. When HNE cells cultures in ALI, average ciliated surface area was 8.6%. CONCLUSION: Our study demonstrated increased ciliogenesis in ALI. PMID- 15362685 TI - [Partial middle turbinatectomy and folded for nasonsociliary neuralgia by transnasal endoscopic surgery]. AB - OBJECTIVE: To explore into the feasibility and outcome of partial middle turbinetectomy and folded for nasociliary neuralgia by transnasal endoscopic surgery. METHOD: Forty patients with nasociliary neuralgia were treated by partial middle turbinatectomy and folded. RESULT: All patients have got sufficient of factory sulcus without hyposmia. The curable rate and the improve rate were 97.5% and 2.5%, respectively, with the total effective rate 100%. CONCLUSION: This procedure present an ideal effect. It is a safe, minimally invasive and efficient procedure for nasociliary neuralgia. PMID- 15362686 TI - [The expressions of Ki67 and VEGF in squamous cell carcinoma of larynx and the correlation between the two marks]. AB - OBJECTIVE: To investigate the expression of Ki67 and VEGF in squamous cell carcinoma of larynx and the correlation between Ki67 and VEGF. METHOD: The expression of Ki67 and VEGF in squamous cell carcinoma of larynx tissues of 40 cases were studied by immunohistochemical staining. RESULT: The expression of Ki67 and VEGF in squamous cell carcinoma of larynx tissues were higher than those in mucosa of laryngeal cavity (P<0.01). The expression of the two marks in squamous cell carcinoma of larynx with the metastasis of lymph nodes were higher than those without metastasis (Ki67: P<0.05, VEGF: P<0.01). Age and sex did not change the expression obviously (P>0.05). There was a positive correlation between T-stage and immunohistochemical staining (P<0.05). There was also a positive correlation in staining intensity between Ki67 and VEGF (r = 0.823, P<0.01). CONCLUSION: Ki67 and VEGF are objective marks to estimate the behaviors of squamous cell carcinoma of larynx, and they can do help to the diagnosis, the judgment of stages and the treatment of carcinoma of larynx. PMID- 15362687 TI - [The T-lymphocytes subsets expression in the human recurrent nasal polyps]. AB - OBJECTIVE: To investigate the expression of T-lymphocyte subsets (CD4+, CD8+ and CD45RO+ cells) in human recurrent nasal polyps. METHOD: Nasal polyps tissue samples and peripheral blood were obtained from 17 patients, normal human inferior turbinate mucosa and peripheral blood were obtained as well. Flow cytometry was adopted to detect the expression of surface markers of the T lymphocytes. All data were analyzed with t-test. RESULT: There were significantly large number of CD4+, and CD45RO+ cells in the tissue of recurrent nasal polyps. The expression percentage of CD3+ CD4+ cells was significantly higher than that of CD3+ CD8+ cells. Ratio of CD3+ CD4+/CD3+ CD8+ was 1.956 +/- 0.093. CONCLUSION: There were generous of T-lymphocytes expression in human recurrent nasal polyps. The ratio of the T-lymphocytes subsets was abnormal high than the usual and this indicated the immunological function were in disorder in the local area. The abnormally higher expression of CD3+ CD4+ cells in the local area may play an important role in the recurrence of human nasal polyps. PMID- 15362688 TI - [Relationship among expression of the VEGF gene and MVD with cervical lymph nodes metastasis in laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To study the relation among the expression of the vascular endothelial growth factor (VEGF) gene and microvessel Density (MVD) with cervical lymph nodes metastasis in laryngeal squamous cell carcinoma (LSCC). METHOD: Appling semi-RT PCR technique and immunohistochemistry detected the expression of VEGFmRNA and protein and MVD count in 60 cases with LSCC. RESULT: The expression of VEGF and MVD was both higher either at molecular level or at the protein level in the group with cervical lymph nodes metastasis than the those with cervical lymph nodes-negative metastasis. The overexpression of VEGF had a positive correlation with the higher MVD count and were both related to cervical lymph nodes metastasis in the 60 cases with LSCC. CONCLUSION: There was a significant correlation between the expression of VEGF and MVD, which suggested that the overexpression of VEGF was essential to microangiogenesis of LSCC, and the both were related to cervical lymph nodes metastasis and would become a biologic indexes to prognosis of LSCC. Antiangiogenic therapy with specific antibodies to VEGF leads to inhibit angiogenesis and prevent tumor from growth and metastasis, which might have a bright future in clinical application. PMID- 15362689 TI - [A morphology study on normal adult nasal appearance]. AB - OBJECTIVE: This study is to explore figure characteristic of normal adult nose. METHOD: Three hundreds (150 males and 150 females, 17-25 years old) students were analysized with human measurement of nasal appearance. RESULT: Fifteen means and seven symmetric indexes were acquired. Normal standard data were made. CONCLUSION: The measurement results can give some morphology data to nasal aesthetic plastic surgery and give some important reference to treatment and evaluation. PMID- 15362690 TI - [Expression of Bcl-2 mRNA and Bcl-2 protein in nasal mucusa of rat in allergic rhinitis model]. AB - OBJECTIVE: To study the expression of Bcl-2 (B-cell lymphoma/leukemia-2) mRNA and Bcl-2 protein in nasal mucusa of rat in allergic rhinitis (AR) model. METHOD: Twenty male or female SD rats, weighing 180-220 g, free of nasal diseases, were randomly divided into two groups. AR model was established through intraperitoneal injection of ovabumin (OVA) as well as nasal challenge with OVA. The nasal mucusa thus obtained from two groups were studied by HE, in situ hybridization histochemical staining of Bcl-2 mRNA and immunohistochemical staining of bcl-2/bax protein. RESULT: The positive expression of Bcl-2 mRNA and Bcl-2/bax protein was presented in the glandular cells,epithelium cells and eosinophils. The staining density of Bcl-2 mRNA and Bcl-2 protein in AR rats increased significantly as compared with those in normal controls. No significant differences of Bax protein expression in two groups were found. CONCLUSION: Bcl-2 mRNA and Bcl-2 protein were up-regulated in the mucusa of AR rats. PMID- 15362691 TI - [Experimental and clinical study on intraoperative monitoring of facial nerve]. AB - OBJECTIVE: The aim of the study is to determine parameters that be predictive of function of facial nerve after facial nerve decompression surgery. METHOD: Fifteen New Zealand white rabbits (30 facial nerves) and 21 patients with peripheral facial paralysis were monitored intraoperatively during facial nerve decompression surgery. These patients were followed up for at least 6 months. RESULT: Threshold of EMG is a steady parameter of assessing the function of facial nerve. The threshold between different segments of facial nerve has no difference in New Zealand white rabbits. That there exist EMG events were correlated significantly with good postoperative facial nerve function. CONCLUSION: Threshold of EMG can be used as objective and steady parameter that assess the function of facial nerve. Intraoperative monitoring events can be used for assessment of postoperative facial nerve function. PMID- 15362692 TI - 1800 MHz radiofrequency (mobile phones, different Global System for Mobile communication modulations) does not affect apoptosis and heat shock protein 70 level in peripheral blood mononuclear cells from young and old donors. AB - PURPOSE: To study if prolonged in vitro exposure to 1800MHz radiofrequency (RF) could exert an effect on human peripheral blood mononuclear cells (PBMC) from young and elderly donors by affecting apoptosis, mitochondrial membrane potential and heat shock protein (HSP) 70 levels. MATERIALS AND METHODS: Endpoints were analysed in the presence or absence of the apoptosis-inducing agent 2-deoxy-D ribose. Three different signal modulations typical of the Global System for Mobile communication (GSM) system were applied. The modulations are widely used in mobile telephony (GSM Basic, discontinuous transmission [DTX] and Talk) at specific absorption rates of 1.4 and 2.0 W kg(-1). RESULTS: In all conditions and for all endpoints tested, there was no significant difference between RF- and sham-exposed cells. CONCLUSION: 1800MHz RF could not induce apoptosis by itself or affect the apoptotic phenomenon when induced by an apoptotic agent. Moreover, RF did not modify the mitochondrial functionality and the expression of HSP 70. PMID- 15362693 TI - Persistence of altered 5-hydroxytryptamine turnover following hemibody X irradiation in the rat distal colon. AB - PURPOSE: Acute gastrointestinal responses to ionizing radiation exposure include a role for 5-hydroxytryptamine (5-HT), but it is not known whether involvement of 5-HT persists and contributes to late effects. The aim was to investigate the acute and later effects of lower hemibody irradiation on 5-HT turnover and the biological effect in the rat distal colon. MATERIALS AND METHODS: Rats were exposed to 10 Gy lower hemibody X-radiation. 5-HT and 5-hydroxyindole acetic acid tissue levels were measured in the distal colon along with the serotonin re uptake transporter and tryptophan hydroxylase mRNA. 5-HT-containing cells and crypt cell numbers were estimated in addition to 5-HT-stimulated short-circuit current responses in isolated mucosa. Studies were performed from 3 days to 3 months post-exposure. RESULTS: During the acute phase, at 3 days post irradiation, reductions in cell number, tissue resistance, serotonin re-uptake transporter expression and secretory responses to 5-HT were observed. However, at later times when secretory responses were normal, 5-HT tissue levels and enterochromaffin cell numbers were increased. CONCLUSIONS: The results provide evidence that after 10 Gy hemibody irradiation, modifications persist past the acute phase. In particular, 5-HT turnover in the distal colon is altered during a longer period. PMID- 15362694 TI - Targeting of diethylene triamine pentaacetic acid encapsulated in liposomes to rat liver: an effective strategy to prevent bone deposition and increase urine elimination of plutonium in rats. AB - PURPOSE: To modify the distribution of the chelating agent diethylene triamine pentaacetic acid (DTPA) by using a formulation approach with liposomes in order to match the in vivo distribution of plutonium (Pu) and, as a consequence, to improve actinide decorporation. MATERIALS AND METHODS: DTPA was encapsulated in conventional and stealth liposomes. Their pharmacokinetics and ability to remove Pu were evaluated in rats 2 and 16 days after a single intravenous treatment given 2 h after contamination with colloidal Pu (239Pu phytate) or with soluble Pu (238Pu citrate). RESULTS: Both formulations induced major pharmacokinetic modifications in rats, allowing an accumulation of [14C]-DTPA mainly in the liver and secondarily (for stealth liposomes) in bone and spleen. These modifications were associated with major increases in urine elimination and with a decrease in skeletal Pu deposition, depending of the nature of the Pu contaminant. After contamination by Pu phytate, conventional liposomes of DTPA (6 micromol kg(-1)) were as efficient as free DTPA (30 micromol kg(-1)) in maintaining the Pu content in the femur below 4.3% of the injected dose after 16 days, a 3.6-fold reduction compared with free DTPA (4 micromol kg(-1)) treatment or without treatment. CONCLUSIONS: A formulation approach with liposomes appears to be a powerful tool to improve the efficiency of Pu chelating agents in vivo. PMID- 15362695 TI - Induction of reproductive cell death and chromosome aberrations in radioresistant tumour cells by carbon ions. AB - PURPOSE: To study the induction of reproductive cell death and chromosome aberrations in radioresistant tumour cells exposed to carbon ions in vitro. MATERIALS AND METHODS: X-ray-resistant colon carcinoma cells (WiDr) were used. Confluent G0/G1 cells were irradiated in vitro with graded doses of 100/200/400 MeV u(-1) carbon ions and carbon ions from the middle of a 1 cm extended Bragg peak, and 200 kV X-rays for comparison. Cells were harvested in their first post irradiation division and aberrations were analysed either by the Giemsa/Hoechst 33258-staining technique or by the fluorescent in-situ hybridization technique involving whole chromosome hybridization and 4',6-diaminido-2-phenylidole (DAPI) staining. Whole chromosome probes were used for chromosomes 2, 4 and 5, and the chromosome painting patterns were classified according published protocols. Reproductive cell survival was determined by a standard clonogenic assay. RESULTS: With respect to the induction of reproductive cell death and chromosome aberrations, carbon ions of different energies were more effective than 200 kV X rays. As expected, irradiation in the extended Bragg peak was the most efficient mode. For cell killing, relative biological effectiveness increased with linear energy transfer up to 2.9. The frequencies of total dicentrics and excess acentric fragments as determined in Giemsa-stained cells were higher in cells irradiated with carbon ions than in cells with X-rays. For 100 MeV u(-1) ions, the dose dependence of apparently simple dicentrics as determined for chromosomes 2, 4 and 5 by single-colour fluorescent in-situ hybridization was linear up to 4 Gy, and linear-quadratic for excess acentric fragments and apparently simple translocations. After irradiation with D=4 Gy carbon ions with energy of 100 MeV u(-1) and from the extended Bragg peak, 12 and 54% of cells displayed complex exchanges, respectively. In contrast, after irradiation with D=4 Gy X-rays, only 1% of cells displayed complex aberrations. Hence, the number of cells with complex exchange aberrations increased strongly after irradiation with carbon ions. CONCLUSION: An increased biological efficiency of carbon ions could be confirmed in radioresistant tumour cells with respect to the induction of reproductive cell death and of unstable as well as stable chromosome aberrations. Relative biological effectiveness reached 2.9 for cell killing by carbon ions from the extended Bragg peak. The yields of apparently simple dicentrics as well as of total dicentrics, i.e. simple dicentrics plus dicentrics belonging to complex exchanges, evaluated in Giemsa-stained metaphases as observed in first post-irradiation mitoses were rather low. In contrast, apparently simple translocations displayed yields systematically higher than simple dicentrics in WiDr cells irradiated with either X-rays or 100 MeV u(-1) or Bragg peak carbon ions. Frequencies o f cells containing complex aberrations increased dramatically after carbon ion irradiation, reaching a maximum for ions from the extended Bragg peak. PMID- 15362696 TI - Distribution of breakpoints and fragment sizes in human chromosome 5 after heavy ion bombardment. AB - PURPOSE: To measure the location of heavy ion-induced residual breakpoints in human chromosome 5 and the size distribution of chromosome fragments involved in inter- or intrachromosomal exchanges. MATERIALS AND METHODS: Human peripheral blood lymphocytes were exposed to 4 Gy accelerated 56Fe (iron) ions (500 MeV per nucleon (MeV n(-1)), linear energy transfer=200 keV microm(-1)). Cells were then stimulated to grow in vitro for 48 h, and chromosomes were prematurely condensed by calyculin A. Chromosome 5 was painted using high-resolution multicolour banding. The location of the observed residual breakpoints and the size of all chromosome 5 fragments involved in structural aberrations were measured using dedicated image analysis software. RESULTS: Mapping of 283 breakpoints revealed a slight deviation from randomness, with an excess of breakpoints clustered in two small bands and an under representation of breaks at the telomeric end in the q arm. Breakpoints per unit length were similar in p- and q-arms. The distribution of chromosome fragments has a maximum for very small fragments (< 10% of the chromosome size), indicating a severe fragmentation of chromosome 5 after heavy ion bombardment. Only fragments < 40% of the chromosome size were involved in intrachromosomal exchanges (interstitial deletions or inversions), whereas fragments up to 75% of the whole chromosome 5 were found in interchromosomal exchanges. CONCLUSIONS: Residual breakpoints after exposure to high-energy iron ions were not distributed randomly along chromosome 5, although the p- and q-arms displayed similar radiosensitivity. Large fragments are either restituted or misrejoined to other chromosome ends, whereas small intrachromosomal fragments can produce either inter- or intrachromosomal exchanges. PMID- 15362697 TI - Ultraviolet exposure of thymocytes: selective inhibition of apoptosis. AB - PURPOSE: To evaluate selective effects of ultraviolet (UV) irradiation on spontaneous and induced apoptosis in freshly extracted mice thymocytes. MATERIALS AND METHODS: Cells were exposed to UV radiation with emission peaks of 365 nm (UVA) exposures of 1620-10200 J m(-2), of 312 nm (UVB) exposures of 34-1620 J m( 2) or of 254 nm (UVC) exposures of 1.5-1620 J m(-2), and incubated for 5.5 h with or without hydrocortisone, phorbol-12-myristate-13-acetate or anti-Fas antibody. Additionally, cells were irradiated with gamma-rays (5 Gy) before UVB exposure (408 J m(-2)) at different times. Apoptosis was quantified by DNA fragmentation. RESULTS: Up to an irradiation of 5000 J m(-2), UVA exposure did not show any effect on thymocyte apoptosis, while at 10200 J m(-2) irradiation, considerable DNA fragmentation was observed. In contrast, UVB and UVC irradiation clearly inhibited natural and cortisone-induced apoptosis. Moreover, UVB inhibited apoptosis triggered by phorbol-12-myristate-13-acetate and gamma-irradiation, but not by anti-Fas antibody. CONCLUSIONS: The response of mouse thymocytes in culture to UV irradiation strongly depends on the wavelength used. It is suggested that either a survival or an apoptotic pathway occurs depending on the physiological state of the cell, spectral composition of the UV light and cell type. The possible involvement of extracellular signal-regulated kinase and stress-activated protein kinase/c-Jun N-terminal kinase in the apoptotic pathway is discussed. PMID- 15362698 TI - Inhibition of the extracellular signal-regulated kinase (ERK) pathway and the induction of radioresistance in rat 3Y1 cells. AB - PURPOSE: Activation of the extracellular signal-regulated kinase (ERK) pathway generally results in stimulation of cell growth and confers a survival advantage. However, the potential involvement of this pathway in cellular radiosensitivity remains unclear. The study was designed to examine whether the ERK pathway affects intrinsic radiosensitivity in mammalian cells. MATERIALS AND METHODS: Exponentially growing rat 3Y1 cells were used. A specific inhibitor of mitogen activated ERK kinase (MEK), PD98059, was used to inhibit the ERK pathway. In addition, kinase-deficient MEK was expressed in cells to inhibit the pathway in a dominant-negative manner. Activation of ERK was visualized by Western blot using an antibody that recognizes the phosphorylated form of ERK. Radiosensitivity was evaluated by a colony-forming assay. RESULTS: 3Y1 cells treated with PD98059 exhibited a significant inhibition of cell proliferation and radiation-induced transient activation of ERK. Unexpectedly, it was found that the inhibitor enhanced clonogenic radioresistance. This effect on radioresistance was confirmed by expression of kinase-deficient MEK. Apoptotic activities following irradiation were significantly inhibited in PD98059-treated cells as determined by caspase-3 like activities. CONCLUSION: Activation of the MEK/ERK pathway increases clonogenic radiosensitivity in rat 3Y1 cells. These findings, together with a variety of other data, suggest that there might be clinical implications in targeting the MEK/ERK pathway in radiotherapy. PMID- 15362699 TI - Workshop on biological applications of X-ray microscopy and imaging. Advanced Photon Source, Argonne National Laboratory, Argonne, IL, USA, 28-29 April 2003. PMID- 15362700 TI - Publicizing scientific misconduct and its consequences. PMID- 15362701 TI - Scientific information and uncertainty: challenges for the use of science in policymaking. AB - Science can reinforce the healthy aspects of the politics of the policy process, to identify and further the public interest by discrediting policy options serving only special interests and helping to select among "science-confident" and "hedging" options. To do so, scientists must learn how to manage and communicate the degree of uncertainty in scientific understanding and prediction, lest uncertainty be manipulated to discredit science or to justify inaction. For natural resource and environmental policy, the institutional interests of government agencies, as well as private interests, pose challenges of suppression, over-simplification, or distortion of scientific information. Scientists can combat these maneuvers, but must also look inward to ensure that their own special interests do not undermine the usefulness of science. PMID- 15362702 TI - Strategies for consulting with the community: the cases of four large-scale genetic databases. AB - Large-scale genetic databases are being developed in several countries around the world. However, these databases depend on public participation and acquiescence. In the past, information campaigns have been waged and little attention has been paid to dialogue. Nowadays, it is important to include the public in the development of scientific research and to encourage a free, open and useful dialogue among those involved. This paper is a review of community consultation strategies as part of four proposed large-scale genetic databases in Iceland, Estonia, United Kingdom and Quebec. The Iceland Health Sector Database and Estonian Genome Project have followed a "communication approach" in order to address public concerns, whereas, UK Biobank and Quebec CARTaGENE have chosen a "partnership approach" to involve the public in decision-making processes. Following a comparison of community consultation strategies, the main concerns of the public are examined as well as the challenges of involving communities. Importantly, reported across all groups is the concern for confidentiality, respect of the individual, transparency, and the donor's right to access to their own result. However, even if researchers demonstrate a willingness to respect values such as fair representation, transparency and accountability, there is still a risk that the public will mistrust researchers and simply will not participate in sufficient numbers. Complications may arise when individual and community interests conflicts. The implementation of a partnership approach is definitely involving and costly; however, if used properly, this method can improve both participation and so database development. PMID- 15362703 TI - The role of databank managers as guardians of public interests. Commentary on "Strategies for consulting with the community: the cases of four large-scale databanks" (Godard et al.). PMID- 15362704 TI - Consulting the community: limits and expectations. Commentary on "Strategies for consulting with the community: the cases of four large-scale databanks" (Godard et al.). PMID- 15362705 TI - Scientific misconduct and findings against graduate and medical students. AB - Allegations of scientific misconduct against graduate students appear to have unique attributes in the detection, investigation, processes used and sanctions imposed vis-a-vis other populations against which misconduct is alleged and found. An examination of the cases closed by the Department of Health and Human Services' Office of Research Integrity and the National Science Foundation reveals that most of the allegations made against graduate and medical students are for falsification and fabrication. Further, additional processes are used in these cases, e.g., student judicial processes, more students are "set up" and more students admit misconduct. Finally, the sanctions imposed when a finding is made typically involve separation from the institution and the federal sanction ranges from none to debarment. Drawing upon the teachings and circumstances of cases involving graduate student peers is a good vehicle for illustrating the concepts and perils of misconduct to graduate students. PMID- 15362706 TI - Authorship in a small medical journal: a study of contributorship statements by corresponding authors. AB - The authorship criteria of the International Committee of Medical Journal Editors (ICMJE) are widely accepted in biomedical journals, but many studies in large and prestigious journals show that a considerable proportion of authors do not fulfill these criteria. We investigated authorship contributions in a small medical journal outside the scientific mainstream, to see if poor adherence to authorship criteria is common in biomedical journals. We analyzed statements on research contribution, as checked by the corresponding author, for individual authors of 114 research articles, representing 475 authors, submitted to the Croatian Medical Journal (CMJ) from 1999 to 2000. Only 40% of authors fulfilled the ICMJE authorship criteria. The authors listed first on the by-line were more likely to fulfill the authorship criteria than all other authors on the by-line. The percentage of authors fulfilling the ICMJE criteria of authorship decreased with the increase in the number of authors listed on the by-line. These results indicate that poor adherence to ICMJE authorship criteria is poor across biomedical journals, regardless of the size of the scientific community. Authorship and contributorship in biomedical journals, as well as editorial ethical responsibilities towards authorship criteria need critical redefinition and education of both editors and authors. PMID- 15362707 TI - Internationalizing professional codes in engineering. AB - Professional engineering societies which are based in the United States, such as the American Society of Mechanical Engineers (ASME, now ASME International) are recognizing that their codes of ethics must apply to engineers working throughout the world. An examination of the ethical code of the ASME International shows that its provisions pose many problems of application, especially in societies outside the United States. In applying the codes effectively in the international environment, two principal issues must be addressed. First, some Culture Transcending Guidelines must be identified and justified. Nine such guidelines are identified Second, some methods for applying the codes to particular situations must be identified Three such methods are specification, balancing, and finding a creative middle way. PMID- 15362708 TI - Establishing sustainable strategies in urban underground engineering. AB - Growth of urban areas, the corresponding increased demand for utility services and the possibility of new types of utility systems are overcrowding near surface underground space with urban utilities. Available subsurface space will continue to diminish to the point where utilidors (utility tunnels) may become inevitable. Establishing future sustainable strategies in urban underground engineering consists of the ability to lessen the use of traditional trenching. There is an increasing interest in utility tunnels for urban areas as a sustainable technique to avoid congestion of the subsurface. One of the principal advantages of utility tunnels is the substantially lower environmental impact compared with common trenches. Implementing these underground facilities is retarded most by the initial cost and management procedures. The habitual procedure is to meet problems as they arise in current practice. The moral imperative of sustainable strategies fails to confront the economic and political conflicts of interest. Municipal engineers should act as a key enabler in urban underground sustainable development. PMID- 15362709 TI - The morality of weapons research. AB - I ask whether weapons research is ever justified. Weapons research is identified as the business of the engineer. It is argued that the engineer has responsibility for the uses to which the tools that he designs can be put, and that responsibility extends to the use of weapons. It is maintained that there are no inherently defensive weapons, and hence there is no such thing as 'defensive' weapons research. The issue then is what responsibilities as a professional the engineer has in regard to such research. An account is given to ground the injunction not to provide the means to harm as a duty for the engineers. This account is not, however, absolutist, and as such it allows justifiable exceptions. The answer to my question is thus not that weapons research is never justified but there must be a strong assurance that the results will only be used as a just means in a just cause. PMID- 15362710 TI - A utilitarian argument against torture interrogation of terrorists. AB - Following the September 2001 terrorist attacks on the United States, much support for torture interrogation of terrorists has emerged in the public forum, largely based on the "ticking bomb" scenario. Although deontological and virtue ethics provide incisive arguments against torture, they do not speak directly to scientists and government officials responsible for national security in a utilitarian framework. Drawing from criminology, organizational theory, social psychology, the historical record, and my interviews with military professionals, I assess the potential of an official U.S. program of torture interrogation from a practical perspective. The central element of program design is a sound causal model relating input to output. I explore three principal models of how torture interrogation leads to truth: the animal instinct model, the cognitive failure model, and the data processing model. These models show why torture interrogation fails overall as a counterterrorist tactic. They also expose the processes that lead from a precision torture interrogation program to breakdowns in key institutions-health care, biomedical research, police, judiciary, and military. The breakdowns evolve from institutional dynamics that are independent of the original moral rationale. The counterargument, of course, is that in a society destroyed by terrorism there will be nothing to repair. That is why the actual causal mechanism of torture interrogation in curtailing terrorism must be elucidated by utilitarians rather than presumed PMID- 15362711 TI - An approach for harmonizing engineering and science education with humaneness. AB - The world is facing an apparently increasing dose of violence. Obviously, there cannot be a simple solution to this complex problem. But at the same time it may be appreciated that, in the interests of humanity, a solution must be pursued in every possible way by everyone. This article is concerned with what one could possibly do at the academic level. Since lack of openness of thought appears to be a fundamental contributor to this unfortunate problem, attempting to cultivate this quality at all levels can perhaps go a long way towards making our earth a better place to live in. With science and engineering education, how can one possibly blend this concern? History of science and the subject of measurement uncertainty may present the necessary scope to the educator to discuss with the students the desirability and necessity of this quality. PMID- 15362712 TI - Human genetic testing under examination by the European Union. Report on a citizens' and stakeholders' conference on "human genetic testing, what implications?". PMID- 15362713 TI - Multiplexed cytokine protein expression profiles from spreading depression in hippocampal organotypic cultures. AB - Cytokines are involved in ischemic tolerance, including that triggered by spreading depression (SD), yet their roles in neuroprotection remain incompletely defined. The latter may stem from the pleiotropic nature of these signaling molecules whose complexities for interaction might be better deciphered through simultaneous measurement of multiple targeted proteins. Accordingly, the authors used microsphere-based flow cytometric immunoassays and hippocampal organotypic cultures (HOTCs) to characterize the magnitude, time course, and diversity of cytokine (interleukin [IL] 1alpha, IL-1beta, IL-2, IL-4, IL-6, IL-10, granulocyte macrophage colony-stimulating factor [GM-CSF], interferon-gamma [IFN-gamma], and tumor necrosis factor-alpha [TNF-alpha]) response to SD. GM-CSF was not detected in HOTCs or media. However, SD triggered a significant, generalized increase in seven cytokines evident in HOTCs 6 hours later, with the remaining cytokine, IL 1beta, becoming significantly different at 1 and 3 days. Additionally, these changes extended to include surrounding media for IL-6 and TNF-alpha by 1 and 3 days. This increase was localized to microglia via immunostaining for IL-1alpha, IL-1beta, and interferon-y. IL-10, although significantly more abundant in HOTCs 6 hours after SD, was significantly less abundant in surrounding media at that time and at 1 day. Finally, the generalized early increase in tissue cytokines later settled to a pattern at 3 days of recovery centering on changes in IL 1alpha, IL-1beta, and TNF-alpha, cytokines capable of modulating ischemic injury. PMID- 15362714 TI - Simplified methods for calculating cerebral metabolic rate of oxygen based on 17O magnetic resonance spectroscopic imaging measurement during a short 17O2 inhalation. AB - It has recently been shown that 17O magnetic resonance (MR) spectroscopic imaging at ultrahigh fields provides a reliable method for measuring CMRO2 during a short period of 17O2 gas inhalation. The mathematical (or complete) model used in the 17O MR method for calculating CMRO2 requires simultaneous measurements of multiple parameters including the concentration of H2 17O produced in the brain tissue from inhaled 17O2 gas (Cb), CBF, and the input function for the H2 17O concentration in the feeding artery (Ca). Both invasive and noninvasive measurements are involved in determining all of these parameters. In this article, two simplified methods are proposed and validated for calculating CMRO2 based on 17O MR measurement(s); the first method requires the measurements of Cb and CBF, but not Ca, and the second method only requires a single noninvasive measurement of Cb. The simplified methods were used to calculate CMRO2 in anesthetized rat brain, and the results were compared with those obtained using the complete model. The results from this work show (1) the validity of the simplified methods for quantifying CMRO2, and (2) the feasibility for establishing a completely noninvasive 17O MR approach for imaging CMRO2 in vivo. PMID- 15362715 TI - Massive inborn angiogenesis in the brain scarcely raises cerebral blood flow. AB - The functional consequences of increased capillary densities in the brain resulting from vascular endothelial growth factor (VEGF165) overexpression are unknown. Therefore, the authors measured local CBF using the iodo-[14C]antipyrine technique in transgenic mice expressing brain-specifically sixfold higher VEGF165 levels and in nontransgenic littermates. To reveal possible compensatory vasoconstriction, CBF was also measured during severe hypercapnia (Paco2 > 130 mm Hg). Simultaneously, local capillary density, perfusion state, and blood-brain barrier permeability were assessed. Using the 2-[14C]deoxyglucose method, metabolic effects of VEGF over-expression could be excluded. In transgenic mice all capillaries showed normal morphology and a tight blood-brain barrier. However, 3% nonperfused capillaries in some brain structures indicate ongoing angiogenesis. Capillary density was drastically increased in transgenic mice in white matter structures (70% to 185%), the dentate gyrus (143%), and caudate nucleus (86%). In all other brain structures investigated, capillary densities were moderately increased by approximately 20%. Normocapnic CBF did not differ between transgenic and nontransgenic mice. During maximal hypercapnic vasodilation, CBF was 20% to 30% higher in transgenic mice, although only in brain structures where capillary density was increased more than twofold. These findings suggest that attenuated CBF in transgenic mice during normocapnia is only partly due to a compensatory vasoconstriction, and that microvascular networks in transgenic brains might be ineffectively constructed. PMID- 15362716 TI - Solid or gaseous circulating brain emboli: are they separable by transcranial ultrasound? AB - High-intensity transient signals (HITS) detected by transcranial Doppler (TCD) ultrasound may correspond to artifacts or to microembolic signals, the latter being either solid or gaseous emboli. The goal of this study was to assess what can be achieved with an automatic signal processing system for artifact/microembolic signals and solid/gas differentiation in different clinical situations. The authors studied 3,428 HITS in vivo in a multicenter study, i.e., 1,608 artifacts in healthy subjects, 649 solid emboli in stroke patients with a carotid stenosis, and 1,171 gaseous emboli in stroke patients with patent foramen ovale. They worked with the dual-gate TCD combined to three types of statistical classifiers: binary decision trees (BDT), artificial neural networks (ANN), and support vector machines (SVM). The sensitivity and specificity to separate artifacts from microembolic signals by BDT reached was 94% and 97%, respectively. For the discrimination between solid and gaseous emboli, the classifier achieved a sensitivity and specificity of 81% and 81% for BDT, 84% and 84% for ANN, and 86% and 86% for SVM, respectively. The current results for artifact elimination and solid/gas differentiation are already useful to extract data for future prospective clinical studies. PMID- 15362717 TI - Changes of dopamine turnover in the progression of Parkinson's disease as measured by positron emission tomography: their relation to disease-compensatory mechanisms. AB - An increase in dopamine turnover has been shown to occur early in Parkinson's disease (PD). This study investigated changes of dopamine turnover as a function of PD duration using the effective distribution volume (EDV) for dopamine, determined by positron emission tomography with 6-[18F]-fluoro-L-dopa, and compared them with changes in dopamine synthesis and storage ability, quantified with the fluorodopa uptake rate constant Ki. Six healthy subjects, 9 early PD patients (PD1), and 13 advanced PD patients (PD2) participated in the study. In the caudate, the Ki and EDV for PD1 were not significantly different from the normal values, whereas in the putamen Ki was 63% of normal and EDV was only 35%. Between PDI and PD2 the decline in EDV was higher than that for Ki (caudate 44% and putamen 46% for EDV vs. 21% and 34%, respectively, for Ki). Turnover was higher in the caudate than the putamen in controls, whereas the PD patients exhibited the reverse pattern. This comparison of changes in Ki and EDV as a function of disease progression indicates that a relatively slower decrease in dopamine synthesis and a relatively faster increase in turnover in early disease likely act as compensatory mechanisms, and that the clinical onset of PD reflects a global failure of dopaminergic compensatory mechanisms. PMID- 15362718 TI - Window of opportunity of cerebral hypothermia for postischemic white matter injury in the near-term fetal sheep. AB - Postresuscitation cerebral hypothermia is consistently neuroprotective in experimental preparations; however, its effects on white matter injury are poorly understood. Using a model of reversible cerebral ischemia in unanesthetized near term fetal sheep, we examined the effects of cerebral hypothermia (fetal extradural temperature reduced from 39.4 +/- 0.1 degrees C to between 30 and 33 degrees C), induced at different times after reperfusion and continued for 72 hours after ischemia, on injury in the parasagittal white matter 5 days after ischemia. Cooling started within 90 minutes of reperfusion was associated with a significant increase in bioactive oligodendrocytes in the intragyral white matter compared with sham cooling (41 +/- 20 vs 18 +/- 11 per field, P < 0.05), increased myelin basic protein density and reduced expression of activated caspase-3 (14 +/- 12 vs 91 +/- 51, P < 0.05). Reactive microglia were profoundly suppressed compared with sham cooling (4 +/- 6 vs 38 +/- 18 per field, P < 0.05) with no effect on numbers of astrocytes. When cooling was delayed until 5.5 hours after reperfusion there was no significant effect on loss of oligodendrocytes (24 +/- 12 per field). In conclusion, hypothermia can effectively protect white matter after ischemia, but only if initiated early after the insult. Protection was closely associated with reduced expression of both activated caspase-3 and of reactive microglia. PMID- 15362719 TI - Dynamic tracking of acute ischemic tissue fates using improved unsupervised ISODATA analysis of high-resolution quantitative perfusion and diffusion data. AB - High-resolution (200 x 200 x 1,500 microm3) imaging was performed to derive quantitative cerebral blood flow (CBF) and apparent diffusion coefficient (ADC) maps in stroke rats (permanent occlusion) every 30 minutes up to 3 hours after occlusion onset, followed by histology at 24 hours. An improved automated iterative-self-organizing-data-analysis-algorithm (ISODATA) was developed to dynamically track ischemic tissue fate on a pixel-by-pixel basis during the acute phase. ISODATA-resolved clusters were overlaid on the CBF-ADC scatterplots and image spaces. Tissue volume ADC, and CBF of each ISODATA cluster were derived. In contrast to the single-cluster normal left hemisphere (ADC = 0.74 +/- 0.02 x 10( 3) mm2/s, CBF = 1.36 +/- 0.22 mL g(-1)min(-1), mean +/- SD, n = 8), the right ischemic hemisphere exhibited three ISODATA clusters, namely: "normal" (normal ADC and CBF), "ischemic core" (low CBF and ADC), and at-risk "perfusion-diffusion mismatch" (low CBF but normal ADC). At 180 minutes, the mismatch disappeared in five rats (Group I, 180-minute "core" lesion volume = 255 +/- 62 mm3 and 24-hour infarct volume = 253 +/- 55 mm3, P > 0.05), while a substantial mismatch persisted in three rats (Group II, 180-minute CBF-abnormal volume = 198 +/- 7 mm3 and 24-hour infarct volume 148 +/- 18 mm3, P < 0.05). The CBF (0.3 +/- 0.09 mL g( 1)min(-1)) of the "persistent mismatch" (Group II, 0.3 +/- 0.09 mL g(-1)min(-1)) was above the CBF viability threshold (0.2 to 0.3 mL g(-1)min(-1)) throughout and its ADC (0.70 +/- 0.03 x 10(-3) mm2/s) did not decrease as ischemia progressed. In contrast, the CBF (0.08 +/- 0.03 mL g(-1)min(-1)) of the analogous brain region in Group I was below the CBF viability threshold, and its ADC gradually decreased from 0.63 +/- 0.05 to 0.43 +/- 0.03 x 10(-3) mm2/s (ADC viability threshold = 0.53 +/- 0.02 x 10(-3) mm2/s). The modified ISODATA analysis of the ADC and CBF tissue characteristics during the acute phase could provide a useful and unbiased means to characterize and predict tissue fates in ischemic brain injury and to monitor therapeutic intervention. PMID- 15362720 TI - Transient focal ischemia affects the cAMP second messenger system and coupled dopamine D1 and 5-HT1A receptors in the living monkey brain: a positron emission tomography study using microdialysis. AB - Using positron emission tomography (PET) and microdialysis, the present study showed that neuronal damages after transient focal ischemia was partly induced by hyperactivation of the cyclic adenosine 3',5'-monophosphate (cAMP) second messenger system through modulations of dopamine D, and serotonin 5-HT1A receptors in the living brains of cynomolgus monkeys. Occlusion of the right middle cerebral artery for 3 hours suppressed CBF in the striatum, and reperfusion induced hyperperfusion in the neocortex and striatum of the occluded side. Six hours after reperfusion, the activity of the cAMP second messenger system assayed with [11C]rolipram was significantly facilitated in the neocortex and striatum where CBF was lowered more than 40% of normal during occlusion ("ischemic" area). Seven days later, impaired dopamine D1 and 5-HT1A receptor binding, measured with [11C]SCH23390 and [carbonyl-11C]WAY-100635, respectively, was observed in the ischemic area. Microdialysis analysis revealed that the striatal dopamine level provided a transient and marked increased during occlusion and after reperfusion, whereas the cortical serotonin level transiently increased only after reperfusion, and was at an undetectable level thereafter. Administration of rolipram (0.1 and 1 mg/kg, intravenously) during occlusion facilitated reduction of dopamine D1 binding, whereas rolipram administration 6 hours after reperfusion induced a further decrease in 5-HT1A receptor binding. These results suggest that the activation of cAMP second messenger system modulated by dopamine D1 and 5-HT1A receptors could be involved in the neuronal degeneration after transient cerebral ischemic insult. PMID- 15362721 TI - Platelet-leukocyte-endothelial cell interactions after middle cerebral artery occlusion and reperfusion. AB - The adhesion of both leukocytes and platelets to microvascular endothelial cells has been implicated in the pathogenesis of ischemia/reperfusion (I/R) injury in several vascular beds. The objectives of this study were to (1) assess the platelet-leukocyte-endothelial cell interactions induced in the cerebral microvasculature by middle cerebral artery occlusion (MCAO)/reperfusion, and (2) define the molecular determinants of the prothrombogenic and inflammatory responses in this model of focal I/R. MCAO was induced for 1 hour in wild-type (WT) mice, WT mice treated with a monoclonal antibody (mAb) to either P-selectin or GPIIb/IIIa, and in P-selectin-/-(P-sel-/-) chimeras. Isolated platelets labeled with carboxyfluorescein diacetate succinimidyl ester (CFDASE) were administered intravenously and observed with intravital fluorescence microscopy. Leukocytes were observed after intravenous injection of rhodamine 6G. One hour of MCAO followed by 1 hour of reperfusion resulted in the rolling and adhesion of leukocytes in venules, and after 4 hours of reperfusion, the adhesion of both leukocytes and platelets was detected. Although both the P-selectin and GPIIb/IIIa mAbs significantly reduced the adhesion of leukocytes and platelets at 4 hours of reperfusion, the antiadhesive effects of the P-selectin mAb were much greater. The leukocyte and platelet adhesion responses were significantly attenuated in both P-sel-/- --> WT and WT --> P-sel-/- bone marrow chimeras, compared with WT --> WT chimeras. Neutropenia, induced by antineutrophil serum treatment, also reduced the recruitment of leukocytes and platelets after cerebral I/R. These findings implicate a major role for both platelet-associated and endothelial cell-associated P-selectin, as well as neutrophils in the inflammatory and prothrombogenic responses in the microcirculation after focal cerebral I/R. PMID- 15362722 TI - Signaling pathways for early brain injury after subarachnoid hemorrhage. AB - Few studies have examined the signaling pathways that contribute to early brain injury after subarachnoid hemorrhage (SAH). Using a rat SAH model, the authors explored the role of vascular endothelial growth factor (VEGF) and mitogen activation protein kinase (MAPK) in early brain injury. Male Sprague-Dawley rats (n = 172) weighing 300 to 350 g were used for the experimental SAH model, which was induced by puncturing the bifurcation of the left anterior cerebral and middle cerebral arteries. The blood-brain barrier (BBB), brain edema, intracranial pressure, and mortality were evaluated at 24 hours after SAH. The phosphorylation of VEGF and different MAPK subgroups (ERK1/2, p38, and JNK) were examined in both the cortex and the major cerebral arteries. Experimental SAH increased intracranial pressure, BBB permeability, and brain edema and produced high mortality. SAH induced phosphorylation of VEGF and MAPKs in the cerebral arteries and, to a lesser degree, in the cortex. PP1, an Src-family kinase inhibitor, reduced BBB permeability, brain edema, and mortality and decreased the phosphorylation of VEGF and MAPKs. The authors conclude that VEGF contributes to early brain injury after SAH by enhancing the activation of the MAPK pathways, and that the inhibition of these pathways might offer new treatment strategies for SAH. PMID- 15362723 TI - Celecoxib induces functional recovery after intracerebral hemorrhage with reduction of brain edema and perihematomal cell death. AB - The selective cyclooxygenase-2 (COX-2) inhibitor has been reported to have antiinflammatory, neuroprotective, and antioxidant effects in ischemia models. In this study, the authors examined whether a selective COX-2 inhibitor (celecoxib) reduces cerebral inflammation and edema after intracerebral hemorrhage (ICH), and whether functional recovery is sustained with longer treatment. ICH was induced using collagenase in adult rats. Celecoxib (10 or 20 mg/kg) was administered intraperitoneally 20 minutes, 6 hours, and 24 hours after ICH and then daily thereafter. Seventy-two hours after ICH induction, the rats were killed for histologic assessment and measurement of brain edema and prostaglandin E2. Behavioral tests were performed before and 1, 7, 14, 21, and 28 days after ICH. The brain water content of celecoxib-treated rats decreased both in lesioned and nonlesioned hemispheres in a dose-dependent manner. Compared with the ICH-only group, the number of TUNEL-positive, myeloperoxidase-positive, or OX42-positive cells was decreased in the periphery of hematoma and brain prostaglandin E2 level was reduced in the celecoxib-treated group. Celecoxib-treated rats recovered better by the behavioral tests at 7 days after ICH throughout the 28-day period, and the earlier the drug was administered, the better the functional recovery. Evidence of similar effects in an autologous blood-injected model showed that direct collagenase toxicity was not the major cause of inflammation or cell death. These data suggest that celecoxib treatment after ICH reduces prostaglandin E2 production, brain edema, inflammation, and perihematomal cell death in the perihematomal zone and induces better functional recovery. PMID- 15362724 TI - Changes in trkB-ERK1/2-CREB/Elk-1 pathways in hippocampal mossy fiber organization after traumatic brain injury. AB - Traumatic brain injury (TBI) leads to mossy fiber reorganization, which is considered to be a causative factor in the development of temporal lobe epilepsy. However, the underlying mechanism is not fully understood. Emerging evidence suggests that TrkB-ERK1/2-CREB/Elk-1 pathways are highly related to synaptic plasticity. This study used the rat fluid-percussion injury model to investigate activation of TrkB-ERK1/2-CREB/Elk-1 signaling pathways after TBI. Rats were subjected to 2.0-atm parasagittal TBI followed by 30 minutes, 4 hours, 24 hours, and 72 hours of recovery. After TBI, striking activation of TrkB-ERK1/2-CREB/Elk 1 signaling pathways in mossy fiber organization were observed with confocal microscopy and Western blot analysis. ERK1/2 was highly phosphorylated predominantly in hippocampal mossy fibers, whereas TrkB was phosphorylated both in the mossy fibers and the dentate gyrus region at 30 minutes and 4 hours of recovery after TBI. CREB was also activated at 30 minutes, peaked at 24 hours of recovery, and returned to the control level at 72 hours of recovery in dentate gyrus granule cells. Elk-1 phosphorylation was seen in CA3 neurons at 4 hours after TBI. The results suggest that the signaling pathways of TrkB-ERK1/2 CREB/Elk-1 are highly activated in mossy fiber organization, which may contribute to mossy fiber reorganization seen after TBI. PMID- 15362725 TI - Plasma ghrelin concentration correlates with the levels of serum pepsinogen I and pepsinogen I/II ratio--a possible novel and non-invasive marker for gastric atrophy. AB - BACKGROUND/AIMS: Ghrelin, a novel growth-hormone-releasing peptide, has been reported to be localized mainly in the A-like cells in the gastric fundic mucosa. With the extension of gastric inflammation caused by H. pylori infection, gastric mucosal atrophy extends from the antrum to the corpus, which is the predominant site of localization of the ghrelin-producing A-like cells. The present study was designed to investigate the correlation between the plasma ghrelin levels and the extent of gastric mucosal atrophy in patients with chronic gastritis caused by H. pylori infection. METHODOLOGY: Sixty-nine patients with dyspeptic symptoms were enrolled for the study. Of these, 41 patients were confirmed to become negative for H. pylori after therapy to eradicate the infection. The other 28 patients were diagnosed as positive for H. pylori infection. Blood samples were collected from all the patients after 12 hours of fasting, before upper gastrointestinal endoscopy was performed. The plasma levels of total and active ghrelin, as well as the serum levels of pepsinogen I (PGI) and pepsinogen II (PGII) were measured by radioimmunoassay. Based on endoscopic assessment, the atrophic changes in the gastric mucosa were classified as open-type atrophy or closed-type atrophy. RESULTS: There were no significant differences in the plasma total and active ghrelin levels between H. pylori-positive and H. pylori-eradicated (negative) patients. The serum levels of PGI correlated well with the plasma levels of total ghrelin (p<0.01, r=0.38) and active ghrelin (p<0.05, r=0.29). The ratio of serum PGI to PGII level (PG I/II ratio) also correlated well with the plasma level of total ghrelin (p<0.05. r=0.31) and active ghrelin (p<0.05, r=0.27). The plasma levels of total as well as active ghrelin were significantly decreased in patients with low PG levels as compared with those in patients with high PG levels (PGI > 70 ng/mL or PGI/II >3.0). The plasma levels of total as well as active ghrelin were also significantly decreased in patients with endoscopically diagnosed open-type atrophy as compared with those in patients with endoscopically diagnosed closed-type atrophy (p < 0.01), especially in the H. pylori-eradicated cohorts. CONCLUSIONS: The plasma levels of ghrelin, which correlated well with the serum levels of PGI as well as the PGI/II ratio, decreased with increasing extent of gastric mucosal atrophy, suggesting that it could be a potentially useful non-invasive marker for chronic atrophic gastritis. PMID- 15362726 TI - Pancreas-preserving biliary amputation with pancreatic diversion: a new surgical technique for complete resection of the intrapancreatic biliary system. AB - Pancreatoduodenectomy is not optimal for organ preservation in patients with mucosal carcinoma of the choledochus. When the lesion spreads near the papilla of Vater, pancreas-preserving biliary amputation may be indicated to achieve complete resection of the biliary system. The first successful case is reported here with technical considerations. First, the pancreatic neck was divided and a tube was inserted into the main pancreatic duct beyond the papilla. The choledochus was dissected downward with division of the posterior pancreatoduodenal vessels. The main pancreatic duct was isolated with the aid of palpation of the tube, and was then ligated and divided. Subsequent dissection was performed to the level of the duodenal mucosa, which was incised circularly. The duodenal defect was then closed. The elevated jejunum was interposed between the pancreatic stumps and bilateral pancreaticojejunostomies were created. The procedure was successfully performed in a patient with superficially spreading cholangiocarcinoma. Postoperative bile leak and pancreatic fistula were controlled with medical management. The patient is currently well without tumor recurrence 19 months after surgery. Her glucose tolerance, which was moderately impaired preoperatively, has been maintained. Pancreas-preserving biliary amputation has been developed as an organ-preserving procedure alternative to pancreatoduodenectomy. Indications, methods of pancreatic reconstruction, and long-term results require further study. PMID- 15362727 TI - Surgery for hepatocellular carcinoma with tumor thrombus extending into the right atrium: report of a successful resection without the use of cardiopulmonary bypass. AB - Hepatocellular carcinomas, of which the tumor thrombus extends into the right atrium via the inferior vena cava, may soon cause fatal complications. Only surgery can be an effective treatment. This procedure usually needs the aid of cardiopulmonary bypass. We recently experienced a successful surgery to remove thrombus combined with hepatectomy. Reporting the detailed technique, both associated diagnosis and intraoperative management are discussed herein. We were able to perform hepatectomy of tumor thrombus in the right atrium without the use of cardiopulmonary bypass or veno-venous bypass. The tumor thrombus was removed from the right atrium into the suprahepatic inferior vena cava by reducing the liver on the tail side. And after total hepatic vascular exclusion was achieved, the intracaval tumor thrombus and the right lobe of the liver were removed en bloc. The operation took 545 minutes and the total hepatic vascular exclusion period was 32 minutes. The postoperative course was uneventful. There are some key points for this procedure. Preoperative or intraoperative US is essential in judging whether tumor thrombus can be removed from the right atrium into the inferior vena cava by reducing the liver or not. Test clamping of the inferior vena cava prior to total hepatic vascular exclusion will enable us to judge whether veno-venous bypass during total hepatic vascular exclusion is needed or not. Surgery without the use of cardiopulmonary bypass is safe and can be minimally invasive when it is performed with a reliable diagnosis and technique. PMID- 15362728 TI - Early removal of bile duct stones in patients with acute biliary pancreatitis by endoscopic papillary balloon dilatation. AB - BACKGROUND/AIMS: Endoscopic papillary balloon dilatation has been accepted as a novel alternative to endoscopic sphincterotomy for the management of bile duct stones. Hence, little or no attempt was made to retrieve stones in cases with acute biliary pancreatitis by endoscopic papillary balloon dilatation. METHODOLOGY: The present study was conducted in ten patients with acute biliary pancreatitis associated with cholestasis or biliary infection. Two patients came with disseminated intravascular coagulopathy, one Child C liver cirrhosis, and another with prolonged prothrombin time of unknown etiology. After the papilla was dilated with a balloon-tipped catheter, the stones were removed with either a retrieval basket catheter, a retrieval balloon, or both. RESULTS: Clearance of the common bile duct was achieved in all ten patients without any serious complications such as pancreatitis aggravation or hemorrhage. Clinical signs and laboratory findings were strikingly improved in all patients. CONCLUSIONS: The results suggest that bile duct stones can be effectively and safely removed by means of endoscopic papillary balloon dilatation even in patients with acute pancreatitis. PMID- 15362729 TI - Evaluation of bilioenteric anastomosis using quantitative hepatobiliary scintigraphy. AB - BACKGROUND/AIMS: The patterns of quantitative hepatobiliary scintigraphy for bilioenteric anastomoses have not been objectively defined. This study was undertaken to establish the patterns of quantitative hepatobiliary scintigraphy in the patients with bilioenteric anastomoses performed for repair of postcholecystectomy benign biliary strictures. METHODOLOGY: 37 patients with bilioenteric anastomosis (Study group) and 10 postcholecystectomy healthy subjects (Controls) underwent quantitative hepatobiliary scintigraphy. Study group patients were further categorized into: Group A (n=27) - normal clinical and biochemical parameters, and Group B (n=10) - abnormal clinical and/or biochemical parameters. On scintigraphy, time of maximal activity and time of clearance of half of the activity was calculated at the liver parenchyma and hepatic hilum. Time of appearance of activity in the intestine was also recorded. RESULTS: There was no significant difference in the scintigraphic parameters between Group A and Controls except for earlier appearance of activity in the intestines (p=0.036) in Group A. In Group B there was significant increase in the time of clearance of half of the activity at the liver parenchyma and hepatic hilum compared to Controls (p=0.003 and 0.036 respectively), and at the liver parenchyma compared to Group A (p=0.002). CONCLUSIONS: Quantitative hepatobiliary scintigraphic patterns in patients with bilioenteric anastomosis were similar to those of postcholecystectomy controls. Patients with abnormal biochemical parameters had significant delay in clearance of activity. Significance of these scintigraphic patterns in this subset of patients can be determined only on long term follow-up. PMID- 15362730 TI - Prevalence of cholesterol gallstones positively correlates with per capita daily calorie intake. AB - BACKGROUND/AIMS: Gallstone composition has changed over the past decades in East Asian countries with a prominent increase in the prevalence of cholesterol gallstones. This trend is possibly due to the westernization of dietary habits. This study was undertaken to delineate the changing trends in gallstone composition in Japan since the 1920s and to assess dietary influences on gallstone composition. METHODOLOGY: Between 1971 and 1999, 1264 Japanese patients underwent cholecystectomy for gallstone disease in Niigata University Medical Hospital. Gallbladder stones retrieved from each patient were classified into four types by gross inspection of the cut surface and infrared spectroscopy: cholesterol stone, black pigment stone, brown pigment stone, and other stones. The literature was reviewed to find both changing trends in gallstone composition before 1971 and dietary changes since 1950 in Japan. RESULTS: The prevalence of cholesterol gallstones among Japanese patients undergoing cholecystectomy increased steadily until the 1970s and declined thereafter (P<0.001). Among dietary factors, only per capita daily total calorie intake strongly correlated with cholesterol gallstone prevalence (r=0.93, P=0.02). CONCLUSIONS: During the 20th century, the prevalence of cholesterol gallstones in the Japanese population increased steadily until the 1970s declining thereafter. Per capita daily total calorie intake appears to be the predominant dietary factor affecting cholesterol gallstone prevalence. PMID- 15362731 TI - Carcinoma of the ampulla of Vater: results of surgical treatment of a single center. AB - BACKGROUND/AIMS: The aim of this study was to retrospectively evaluate our experience with ampulla of Vater tumors at the Creighton University and to establish the role of curative pancreaticoduodenectomy on the long-term survival rate of patients. METHODOLOGY: Between 1975 and 1997, 21 patients (15 M, 6 F) with ampulla of Vater tumors were identified using our tumor registry database. The mean age at diagnosis was 74 years (range 45-85). Family history of other types of cancers was positive in 11 patients. The two most common presenting symptoms were painless jaundice (75%) and abdominal pain (31%). RESULTS: Thirteen patients (62%) underwent curative pancreaticoduodenectomy and eight patients (38%) underwent palliative biliary bypass procedures. Pathological staging of thirteen pancreaticoduodenectomy patients included stage I, 5 patients; stage II, 1 patient; stage III, 2 patients and stage IV, 5 patients. There were nine patients with NO disease and 4 patients with N1 disease. The Kaplan-Meier analysis revealed significant survival benefit for pancreaticoduodenectomy patients (69.9 months) compared to the palliative procedure (4.8 months). Furthermore NO patients had a far better survival (69.9 months) compared to N1 patients (5.9 months) although this difference was not significant. There were no operative mortalities. CONCLUSIONS: Radical surgery such as pancreaticoduodenectomy seems to confirm long-term survival in patients with ampulla of Vater tumors especially when no nodal or distant metastases are present at the time of surgery. An aggressive approach, therefore, in such patients is justifiable. PMID- 15362732 TI - Successful reconstruction of segmental or subsegmental bile ducts after resection of hilar bile ducts: short- and long-term results. AB - BACKGROUND/AIMS: Biliodigestive anastomoses to small bile ducts after extensive liver resection or central bile duct remain difficult. The inherent problems associated with this particular procedure have not been widely discussed. In order to demonstrate that reconstruction of segmental or subsegmental bile ducts with mucosa-to-mucosa alignment using transanastomotic stents can reduce morbidity associated with the procedure, we have reviewed our experience with small biliodigestive anastomosis after liver resection. METHODOLOGY: Thirteen patients were operated on from 1995 to 1997. Twelve had malignant diagnoses. A comprehensive standardized technique included the use of short or trans intestinal trans-anastomotic stents and single layer interrupted absorbable sutures with mucosa-to-mucosa alignment. In the patients, in-hospital morbidity and mortality with focus on bile leaks and long-term signs of cholangitis and stricture formation were investigated with a median observation period of 42 months. RESULTS: Eleven underwent extensive liver resections and two had complicated central resections including the bile duct confluence. The procedures included 40 transected bile ducts and 32 anastomoses to the jejunum. One patient died as a result of rupture of a splenic arterial aneurysm caused by a pancreatic pseudocyst. Other complications which occurred in seven patients were intra abdominal fluid collection (n=5), wound infection (n=1), minor bile leakage from the liver cut surface (n=1), and pleural effusion (n=4). Postoperatively four patients, who had local recurrence with jaundice, showed cholangitis, whereas the remaining nine patients showed no signs of cholangitis. CONCLUSIONS: Our technique for reconstructing small bile ducts with mucosa-to-mucosa alignment is considered adequate from the viewpoint of short- and long-term results. PMID- 15362733 TI - Palliation of unresectable periampullary neoplasms. "surgical" versus "non surgical" approach. AB - BACKGROUND/AIMS: A series of 84 patients with unresectable periampullary neoplasms, observed during an eight-year period (1992-1999), is reviewed for a critical analysis of modalities of treatment and clinical outcome. Two different approaches, a preference for "non-surgical" palliation and a preference for "surgical" palliation, in two consecutive periods, are compared. METHODOLOGY: In the first period (1992-1995) endoscopic retrograde cholangiopancreatography with endoprosthesis insertion was performed routinely and, after diagnostic assessment, patients were divided into two groups: those with apparently resectable neoplasms, candidates for surgery, in whom the decision to perform a surgical palliation by biliary bypass was taken intraoperatively, and those with unresectable neoplasms, in whom the endoprosthesis insertion was considered as definitive palliation. Since January 1996, a different approach was started; surgical bypass was considered the palliation of choice and only patients unfit for surgery underwent endoscopic stenting. RESULTS: Evaluation of the results showed that the first strategy was associated with a high incidence of complications and unsatisfactory long-term results, with frequent hospital readmissions, poor quality of residual life and therefore failure of palliation. On the contrary, the latter approach with preference for surgical palliation was associated with lower morbidity and mortality, with persistent relief of symptoms and better quality of life in a larger percentage of patients. CONCLUSIONS: This study provides evidence that, in patients with unresectable periampullary carcinoma, surgical palliation provides better long-term results than endoscopic palliation. The results were also consistent with the advantage of associating a gastric bypass to the biliary bypass, also in the absence of gastric outlet obstruction. PMID- 15362734 TI - Prevention of bile leak after major hepatectomy. AB - BACKGROUND/AIMS: Bile leak remains a serious complication after major hepatectomy. The usefulness of external biliary drainage to prevent intraperitoneal bile leak was studied. METHODOLOGY: Thirty-nine patients who underwent major hepatectomy from April 1997 through June 2000 were studied. The bile leak test was performed to identify and close leaks following the resection. Patients who still had leakage underwent retrograde transhepatic biliary drainage (RTBD) via a tube inserted through a choledochostomy. Patient's backgrounds, incidence of bile leak, and time until resolution of bile leak were compared between patients who did and did not have an RTBD tube placed. RESULTS: Nineteen patients had an RTBD tube (48.7%). Bile leak developed in 4 patients with the tube (21.1%), and in 4 patients without the tube (20.0%) (not significant). However, the time until resolution of bile leak was 13.3 days for patients with the tube and 51.3 days for patients without the tube (p<0.05). Two patients developed local peritonitis when the tube was removed. CONCLUSIONS: Though some patients had bile leakage even with the RTBD tube, use of the RTBD tube decreased the length of time leakage that occurred. RTBD tube drainage should be done routinely with major hepatectomy. PMID- 15362735 TI - Lymphocyte subpopulations in peripheral blood in primary sclerosing cholangitis. AB - BACKGROUND/AIMS: Primary sclerosing cholangitis (PSC) is a chronic disease of autoimmunological etiology, leading to inflammation, destruction and atrophy of the bile ducts. The aim of the study was to determine peripheral lymphocyte B, T, and NK cells in PSC. METHODOLOGY: The estimation of peripheral blood lymphocytes in 17 patients (54+/-12 years old) with PSC was carried out; the control group consisted of 27 subjects (38+/-11 years). The following T lymphocyte subpopulations were determined using duo color flow cytometry: CD3+, CD4+, CD8+, CD3++HLA DR+, B cells CD19+, and NK cells CD16+ +CD56+. RESULTS: In PSC we observed doubled increase in activated T lymphocytes of CD3+ +HLA DR+ phenotype as compared to healthy subjects (7.9% vs. 2.7%, p<0.01) and NK cells (12.6% vs. 10.3%, respectively, p<0.05). There were no significant differences in the composition of CD3+, CD4+, and CD8+. In peripheral blood we noted, in patients with PSC, a decrease in B lymphocytes (11.2% vs. 12.3%, p<0.19). CONCLUSIONS: The examinations showed that activated T (HLA DR+) lymphocytes and NK cells played an important role in development of PSC. PMID- 15362736 TI - Curative resection of a huge bile duct cancer without pancreatoduodenectomy. AB - A 71-year-old woman was admitted to our hospital with obstructive jaundice. Magnetic resonance cholangiography revealed a 9-cm spindle-shaped tumor in the common bile duct. Laboratory examination noted a high concentration of total serum bilirubin. Although the serum concentration of carcinoembryonic antigen was within normal limits, the serum concentration of carbohydrate antigen 19-9 was elevated. We diagnosed the lesion as an extrahepatic bile duct cancer and performed a laparotomy. The anterior wall of the common bile duct was incised, and the tumor was found to have a small base in only the posterior wall of the duct, and spread within the duct was from the common hepatic duct to the intrapancreatic bile duct. Bile duct resection, cholecystectomy with lymphadenectomy and hepaticojejunostomy were performed. Frozen sections of the proximal and distal surgical margins of the bile duct were free of cancer. Macroscopically, the lesion was an expansive polypoid tumor measuring 9x3cm in diameter arising from a 5-mm base. Histologic examination revealed that the tumor was a well-differentiated tubular adenocarcinoma that had infiltrated the fibromuscular layer at its base. Longitudinal spread was more extensive hepatopetally than hepatofugally. The patient is alive and well without any complaints 8 months after surgery. PMID- 15362737 TI - Endocrine tumors of the extrahepatic bile ducts. Pathological and clinical aspects, surgical management and outcome. AB - Endocrine tumors of the extrahepatic bile ducts are an extremely rare clinical entity with only very few cases being reported to date. Two additional cases of extrahepatic endocrine tumors are presented and a review of the literature is analyzed to provide a comprehensive data collection. Thirty-four cases of extrahepatic endocrine tumor, including our 2 cases, have been reported; 16 (47%) being carcinoma. The mean age of presentation was 48.2 years with a male/female ratio of 0.36. Jaundice, the most common symptom, was found in 63% of the patients. The common bile duct and the hilar region were the two most frequent anatomic locations. The mean size of resectable tumors was 2.37cm (0.2- 5.5cm). Metastases were present in 11 patients (32.35%) and local invasion was found only in 6 patients (17.6%). Curative surgery was realized in the majority of the patients. Long-term disease-free survival as long as 20 years was reported when surgery was curative. Endocrine tumors of the extrahepatic duct are exceptional and represent 0.2% of all cancers of these anatomic sites. Curative resection is the most important prognostic factor. Therefore, aggressive surgical therapy with curative intention offers the only chance for cure and has to be considered whenever possible. PMID- 15362738 TI - Hilar bile duct cancer associated with preoperatively undetectable von Meyenburg complex--report of a case. AB - A 56-year-old man was admitted to our hospital with a diagnosis of hilar bile duct cancer. Abdominal ultrasonography, computed tomography and endoscopic retrograde cholangiopancreatography showed no other concomitant disease. Biochemical data showed 0.6 mg/dL of total bilirubin, 104 IU/L of alanine aminotransferase and 469 mg/dL of alkaline phosphatase. Carbohydrate antigen 19-9 was elevated as 112.1 U/mL. Operative findings included a resectable left hilar bile duct cancer and grayish-white nodules 0.3-0.5cm in diameter on the surface of segments 6 and 4 of the liver. Although intrahepatic metastasis of the bile duct cancer was highly suspected under intraoperative US, frozen section was reported to show the small nodules containing multiple biliary hamartomas, so called von Meyenburg complex. Therefore, a left hepatic lobectomy together with resection of the extrahepatic bile duct followed by a Roux-en-Y hepaticojejunostomy was the procedure of choice. His postoperative course was uneventful. The intraoperative findings could have been misdiagnosed due to their similarity to intrahepatic metastasis and intraoperative histology is indispensable to differentiate von Meyenburg complex in this case. The possibility of a preoperative imaging diagnosis for von Meyenburg complex seems to depend on the size of the bile duct structure in each hamartoma. To the best of our knowledge, this is the fourteenth case of bile duct cancer associated with von Meyenburg complex reported in the literature. The following case is being reported because of the rarity of the disease and to stress the importance of intraoperative histology to avoid misdiagnosis as the disseminated disease, particularly when malignant neoplasia is surgically treated. PMID- 15362739 TI - Primary and recurrent retroperitoneal sarcoma: factors affecting survival and long-term outcome. AB - BACKGROUND/AIMS: To analyze treatment and survival in a series of 39 patients with primary or recurrent retroperitoneal sarcoma treated and prospectively followed at a single institution. METHODOLOGY: Between July 1994 and January 2002, 39 patients (20M, 19F; mean age: 56 years, range: 25-77) were evaluated. RESULTS: Thirty-two out of 39 patients (82%) (18 were affected by primary retroperitoneal sarcoma, and 14 by recurrent retroperitoneal sarcoma), were submitted for resection. Twenty-four out of 32 patients (75%) underwent removal of contiguous intra-abdominal organs. Peroperative mortality was nil and significant peroperative complications occurred in 6 cases only (19%). High tumor grade results were a significant variable for a worse survival in all 32 patients (100% 5 years survival for low grade vs. 0% for high grade; P=0.0004). Among 27 radically resected patients, only histology gradeand peroperative blood transfusions affected survival (100% 5-year survival for low grade vs. 24% for high grade; P=0.003); (100% 5-year survival for nontransfused patients vs. 43% for transfused patients; P=0.03). Similar effects were noted for disease-free survival. CONCLUSIONS: Histology grade and peroperative blood transfusions were the only factors which affected overall and disease-free survival. An aggressive surgical approach in both primary and recurrent retroperitoneal sarcoma is associated with long-term survival. PMID- 15362740 TI - The value of p53 protein expression in gallbladder carcinoma: analysis of 60 cases. AB - BACKGROUND/AIMS: Few studies, with small samples and diverging results, have been performed to evaluate the p53 protein expression in gallbladder carcinoma and its relationship to different clinicopathological parameters. Based on these facts, we performed a study for the purpose of assessing p53 expression in this disease and its association to prognostic factors. METHODOLOGY: Samples of 141 gallbladders, with 60 cases of carcinoma, 62 cholelithiasis and 19 without gallstones were assessed using an immunohistochemical technique for the expression of p53 protein, and analyzed for prognosis, survival and other clinicopathological parameters. RESULTS: p53 expression was positive in 58.3% of carcinomas of the gallbladder, 9.7% of the chronic cholecystitis and 10.5% of the gallbladders not associated to stones. In cases of carcinoma of the gallbladder there was no statistically significant association between the expression of this protein, the prognostic factors, histological type or grade, presence of gallstones and survival. CONCLUSIONS: The mutation of gene p53 is involved in the pathogenesis of carcinoma of the gallbladder, and the intense chronic inflammatory process associated or not with cholelithiasis, appears to be one of the factors involved in the genesis of this process. Our data do not show an association between p53 protein expression and patient's survival prognosis. PMID- 15362741 TI - Diagnostic utility of GLUT1 in the differential diagnosis of liver carcinomas. AB - BACKGROUND/AIMS: Malignant cells require high energy via glycolytic generation of ATP for cell proliferation and survival. This process is thought to be mediated by glucose transporters (GLUTs). GLUT1 appears to be expressed aberrantly in many cancers. The liver is the main organ closely related to glycogen metabolism and it is thought that GLUT expression might be altered in the tumors arising in the liver. METHODOLOGY: We studied GLUT1 expression in 22 hepatocellular carcinomas (HCC) and 16 cholangiocarcinomas (CC) and compared it to the expression of hepatocyte specific antigen (HSA) and cytokeratin 19 (CK19) by immunohistochemical studies. RESULTS: In HCC, GLUT1 was expressed in 4.5% (1/22), HSA in 77.3% (17/22) and CK19 in none (0/22). In CC, GLUT1 was expressed in 81.3% (13/16), HSA in none (0/16) and CK19 in 100% (16/16). Interestingly, GLUT1 was not expressed in normal hepatocytes and bile duct epithelium around the tumor, whereas CK19 was expressed in normal bile ducts and tumor cells. CONCLUSIONS: These results indicate that GLUT1 might be related to development of bile duct carcinoma and its expression was a more reliable marker for detection of bile duct carcinoma than CK19. Additionally, GLUT1 might potentially be useful as a means of distinguishing CC from HCC. PMID- 15362742 TI - The expression pattern of vascular endothelial growth factor C and D in human esophageal normal mucosa, dysplasia and neoplasia. AB - BACKGROUND/AIMS: Vascular endothelial growth factor C (VEGF-C) and D (VEGF-D) are considered to be potentially lymphangiogenic and can selectively induce hyperplasia of the lymphatic vasculature. In this study, we examined the expression of VEGF-C and -D in esophageal carcinoma. METHODOLOGY: With immunohistochemical staining using specific antibodies, we classified 26 esophageal carcinoma cases and 11 dysplasia cases. RESULTS: All esophageal carcinomas clearly expressed VEGF-C. In esophageal dysplasia, 9 (82%) lcases were positive for VEGF-C, and 2 (18%) were negative. In contrast, none of the normal esophageal mucosa expressed VEGF-C. Seventeen (65%) of 26 cases of esophageal carcinoma were positively stained for VEGF-D and 7 (35%) were negative. VEGF-D was also positive in 2 (18%) cases of esophageal dysplasia, but in no cases of normal tissue. VEGF-C was detected in all carcinomas and dysplastic lesions that expressed VEGF-D. CONCLUSIONS: Active production of VEGF-C and -D was observed not only in esophageal carcinomas but also in some dysplastic lesions. This finding raises the possibility that VEGF-C and -D might play positive roles in the early stage of esophageal carcinogenesis. PMID- 15362743 TI - Early colon cancers detected by FDG-pet: a report of two cases with immunohistochemical investigation. AB - Increased glucose uptake is one of the metabolic characteristics of tumor cells. 18F-fluorodeoxyglucose (FDG)-positron emission tomography (FDG-PET), a technique that is used widely to study this altered glucose metabolism in tumors, allows the detection of various types of malignancy. We present herein two cases of early colon cancers detected incidentally by FDG-PET. The technique was used as part of the screening examinations for preoperative staging, and for postoperative follow-up. In both cases, the lesions were removed by colonoscopic polypectomy, with no complications. Moreover, we confirmed the existence of altered glucose metabolism in the resected specimen by immunohistochemical staining using an antibody raised against Glut1. Immunohistochemically, Glut1 was expressed in vitro in both of the lesions, supporting the positive FDG-PET result obtained in vivo. To our knowledge, this is the first report to describe in vitro Glut1 expression and in vivo tumor detection using FDG-PET in colorectal carcinoma. PMID- 15362744 TI - Aggressive surgical resection for hepatocellular carcinoma with tumor thrombus extending to inferior vena cava and synchronous pulmonary metastasis. AB - We describe a 66-year-old man having hepatocellular carcinoma with tumor thrombus extending into the inferior vena cava and synchronous pulmonary metastasis. He was referred to Chiba University Hospital on May, 2000, complaining of emaciation. Radiological findings showed a huge hepatocellular carcinoma in the entire right lobe and tumor thrombus extended into the intrapericardial inferior vena cava. He also had a solitary pulmonary metastasis in the left pulmonary lobe (stage IVB). Right hemihepatomy was performed under total hepatic vascular exclusion without cardiopulmonary bypass, and tumor thrombus was completely removed. Thoracoscopic wedge resection of pulmonary metastasis was also performed. The patient had an uneventful postoperative course. Histopathological examination revealed that the tumor was moderately differentiated hepatocellular carcinoma The patient is still alive after 26 months with pulmonary recurrence, but without hepatic recurrence. To our knowledge, there has been no reported case of resection for both hepatocellular carcinoma invading the inferior vena cava and synchronous pulmonary metastasis. In conclusion, aggressive surgical resection for advanced hepatocellular carcinoma concomitant with pulmonary resection may bring about better prognosis in highly selected patients. PMID- 15362745 TI - Liver metastases of a minute rectal carcinoid less than 5mm in diameter: a case report. AB - We report a case of liver metastases of a minute rectal carcinoid less than 5mm in diameter, which was found during the postoperative follow-up course of a stomach cancer patient. For the early stomach cancer, laparoscope-assisted distal gastrectomy with lymph node dissection was performed on August 26, 1998. Later, abdominal CT revealed space-occupying lesions in the liver (S2). Metastatic tumors of the stomach cancer were suspected, but further examination revealed that the lesions were metastatic tumors due to a rectal carcinoid tumor. Ten months later, metastatic carcinoid tumors were found in the liver (S1, S5, S6, S7, S8). Subsegmentectomy of the liver (S7) and microwave coagulation therapy (S1, S5, S6, S8) were performed. Carcinoid tumors metastatic to the lymph nodes, liver, and other areas have been reported, but all were larger than 20mm in diameter. In this case, the primary tumor was less than 5mm in diameter, which is extremely rare. This patient was successfully treated with lateral segmentectomy, subsegmentectomy (S7), microwave coagulation therapy of the liver, and transanal extirpation. She is presently in a good condition and has had no recurrence of the carcinoid tumor from ten months after the last hepatectomy. PMID- 15362746 TI - Sensitivity of immunochemical fecal occult blood test for colorectal flat adenomas. AB - BACKGROUND/AIMS: To assess the sensitivity of an immunochemical fecal occult blood test for colorectal flat adenomas, and to determine the relation between the size of flat adenomas and the results of this test. METHODOLOGY: Eleven colorectal flat adenomas under 1cm, 18 flat adenomas 1cm or larger, 187 polypoid adenomas under 1cm, 42 polypoid adenomas 1cm or larger and 144 healthy controls were investigated. Each subject was tested with an immunochemical occult blood test on two consecutive days, and the accuracy of this test was evaluated. RESULTS: The sensitivity of this test was 18% for flat adenomas under 1cm, 67% for flat adenomas 1cm or more, 19% for polypoid adenomas under 1cm and 60% for polypoid adenomas 1cm or more. Specificity was 95%, showing a significant difference in the sensitivity between flat adenomas 1cm or more and those under 1cm (p<0.05) as well as a significant difference between flat adenomas 1cm or more and polypoid adenomas under 1cm (p<0.001), whereas no significant difference was seen in the sensitivity between flat adenomas 1cm or more and polypoid adenomas 1cm or more. CONCLUSIONS: These results indicate that the sensitivity of immunochemical occult test for flat adenomas was similar to the sensitivity for common polypoid adenomas. The size of adenomas is a more important morphological factor for determining the sensitivity of this fecal occult blood test. PMID- 15362747 TI - Progress in sigmoidectomy for cancer during the last 24 years in our department. AB - BACKGROUND/AIMS: In the treatment for colon cancer, changes may have taken place in areas such as surgical devices, concepts for basic strategies, composition of the medical team, postoperative management and adjuvant chemotherapy. No one knows the net effect of these alterations in treatment on sigmoid colon cancer patients. METHODOLOGY: A total of 147 patients were divided into 2 groups based upon the time of the surgery. Comparisons were made based upon the several parameters, including survival. In addition, to determine the effect of the degree of lymph node dissection on survival, patients were divided into 3 groups based upon the ligated portion of artery. RESULTS: Blood loss significantly decreased. Other factors were almost the same except for survival. In Dukes' A and B, 5-year survival rates were the same (90%). But in Dukes' C, the 5-year survival rate was better in the later period (90%) than in the earlier period (70%). In Dukes' C, the extent of lymph node dissection significantly influenced the survival. CONCLUSIONS: The parameters have not changed very much in these 24 years except for the 5-year survival rates in Dukes' C. The extent of lymph node dissection is probably sufficient just below the root of left colic artery. PMID- 15362748 TI - Clinical features of second other-site primary cancers among sporadic colorectal cancer patients--a hospital-based study of 3,722 cases. AB - BACKGROUND/AIMS: Although synchronous and metachronous colorectal cancers are now well known to all colorectal surgeons, second other-site primary cancers following colorectal cancer are unfamiliar to most colorectal physicians. Knowledge of the clinical characteristics of these second other-site primary cancers becomes important especially during their follow-up surveillance. METHODOLOGY: Information from a computer database was retrieved. Medical records were reviewed and all histopathological data were confirmed. RESULTS: Forty-seven (1.26%) patients developed second other-site primary cancers. Of these patients, 22 (46.8%) patients were diagnosed concordantly while 25 (53.2%) patients were discordant cases. Twelve of 25 (45%) discordant cases were found within one year postoperatively. Regarding the clinical features of these 47 patients, male patients more frequently develop discordant cases (p=0.031). A significantly higher proportion of Dukes' B patients developed second other-site primary cancer than that of patients of other stages (p=0.017). Liver, 8 (17%) cases, was the most common site followed by the prostate, 6 (13%), stomach, 6 (13%), lung, 5 (11%), head and neck cancers, 5 (11%), and kidney, 3 (6.4%). However, there was not a significantly increased risk for these sporadic colorectal patients statistically. CONCLUSIONS: We described clinical features of second other-site primary cancers and emphasized that knowledge of these is important for their postoperative surveillance. PMID- 15362749 TI - Controlled, open, randomized multicenter trial comparing the effects of treatment on quality of life, safety and efficacy of budesonide foam and betamethasone enemas in patients with active distal ulcerative colitis. AB - BACKGROUND/AIMS: There is evidence of a higher quality of life with foams as compared with enemas. The purpose of this study was to assess the effect of treatment with budesonide foam or betamethasone enema on the quality of life and the clinical outcome in patients with distal ulcerative colitis. METHODOLOGY: In an open multicenter trial, patients with active distal ulcerative colitis were randomized to receive 2 mg/50 mL budesonide foam or 5 mg/100 mL betamethasone enema. Primary outcome variable was the change in the mean Life Quality Index. Therapeutic efficacy was determined by clinical activity, endoscopical and histological indices. RESULTS: 38 patients were included in the study. The decrease of the mean Life Quality Index was more pronounced in the budesonide group. No significant difference in the efficacy of treatment was observed for both groups. Betamethasone suppressed the plasma cortisol level in the majority of the patients (87%) compared to only 22% of the patients receiving budesonide. CONCLUSIONS: The quality of life is not significantly different in patients during treatment with budesonide foam or betamethasone enema for active distal ulcerative colitis. However, while having comparable clinical efficacy budesonide foam has less effect on the plasma cortisol level thus potentially minimizing steroid side effects. PMID- 15362750 TI - Management of postoperative follow-up and surgical treatment for Krukenberg tumor from colorectal cancers. AB - BACKGROUND/AIMS: The purpose of this retrospective study was to evaluate the clinical presentation as well as surgical intervention for ovarian metastasis from colorectal cancers identified during postoperative follow-up. METHODOLOGY: Nine cases of ovarian metastasis were observed among 452 female patients with colon cancers between 1990 and 2000. Initial symptoms were pain (67%), pelvic mass (50%), vaginal bleeding (33%), and uterine bleeding (17%). On pathological evaluation, six cases (67%) were found to be moderately differentiated, and three (33%) well differentiated adenocarcinomas. RESULTS: Early diagnosis is very difficult. The growth of metastatic ovarian tumors are slow in elder patients, its growth is rapid in younger patients, and frequently diagnosed as huge ovarian tumors. In some cases, as cystic ovarian lesions they were identified in their early stage but could not be diagnosed as ovarian metastases, later curative operation could not be performed. The consulting gynecologist could not reach the correct diagnosis. Regular postoperative pelvic CT or MRI should be helpful for diagnosis. Although serum CEA levels did not increase in most cases, tumor markers CA125 and SLX were elevated in several cases, and thus may be useful for the detection of ovarian metastases. Surgical treatment consisted of bilateral and unilateral salpingo-oophorectomy or pelvic exenteration and additional hysterectomy for one patient because of association with cervical cancer. The median survival time after the primary operation was 20.8 months, ranging from 3 to 96 months. Peritoneal dissemination and bone metastases were frequently observed as recurrence after these operations. Two cases (non-synchronous solitary metastasis or direct involvement, located in the pelvis) survived more than 5 years (85 and 96 months, respectively), and both patients were treated with pelvic exenteration. CONCLUSIONS: This suggests that in general most cases with ovarian metastasis have a poor prognosis and that radical operation such as pelvic exenteration can improve survival only in cases of recurrent solitary ovarian metastasis or local extended disease, i.e. when the lesion is located only in the pelvis. PMID- 15362751 TI - Hypogastric nerve preservation does not increase local recurrence after surgical treatment of rectal carcinoma. AB - BACKGROUND/AIMS: The main aim of the study was to evaluate whether superior hypogastric plexus and hypogastric nerve can be preserved without increasing local recurrence while performing surgical treatment of rectal carcinoma. METHODOLOGY: This was a retrospective study of 129 patients with rectal carcinoma who underwent curative resection with two types of autonomic nerve-sparing operation. The superior hypogastric plexus and bilateral hypogastric nerves were resected in 61 patients and spared in 68 patients. The pelvic plexus was preserved in all the patients. Local recurrence and survival were compared between two operations. RESULTS: After three years, local recurrence cumulative rates were 13.1% after hypogastric nerve removing operation and 10.3% after hypogastric nerve preserving operation. Distant metastasis and corrected 5-year survival rates were 23.0 and 61.6%, respectively after hypogastric nerve-removing operation, while after hypogastric nerve-preserving operation those were 16.2 and 77.4%, respectively. There were no statistically significant differences in local recurrence, distant metastasis and survival between the two groups. CONCLUSIONS: Hypogastric nerve-preserving operation does not appear to carry an increased risk of local recurrence compared with hypogastric nerve-removing operation after an equivalent follow-up period. PMID- 15362752 TI - The surgical treatment of fecal incontinence with artificial anal sphincter implant. Preliminary clinical report. AB - BACKGROUND/AIMS: Fecal incontinence has a serious impact upon patients' quality of life. Several treatment methods are possible according to the pathophysiology of the disease. METHODOLOGY: Between March 1999 and February 2002 eight artificial anal sphincters (American Medical System - AMS) were implanted in seven patients affected with severe fecal incontinence; in one case the device was positioned in a patient who had previously undergone a Miles' resection. All cases were carefully selected according to appropriate diagnostic evaluation. The follow-up varies between 3 and 40 months. RESULTS: The prosthesis had to be removed in two cases; in one patient infection of the implant area occurred, while in the other case persistent perianal pain due to the presence of the device could not be tolerated by the patient. In the six patients that could be successfully treated with the artificial anal sphincter implant, it dramatically improved their quality of life. CONCLUSIONS: The success of the procedure allows the consideration that the artificial anal sphincter implant is the best treatment for severe fecal incontinence that cannot be solved with conservative therapy. PMID- 15362753 TI - Pathological changes of advanced lower-rectal cancer by preoperative radiotherapy. AB - BACKGROUND/AIMS: In the treatment of lower rectal cancer, we examined the pathological effects of preoperative radiotherapy on intramural spread of tumor and risk factors for local recurrence, including tumor deposit, budding growth of primary tumors, and micrometastasis to lymph nodes. METHODOLOGY: Ninety-four patients who underwent surgery for lower rectal cancer were selected. Forty-seven patients received preoperative radiotherapy, at a total dose of 50 Gy [Rad(+) group], whereas 47 did not [Rad(-) group]. Intramural spread was evaluated between the Rad(+) and Rad(-) groups. For analysis of tumor deposit, budding, and micrometastasis, 25 stage-matched patients were selected in both groups. RESULTS: Intramural spread was significantly less in the Rad(+) group, compared with the Rad(-) group. The Rad(+) group showed a significant decrease in tumor deposit, budding, and micrometastasis, compared with the Rad(-) group. In patients having overt lymph node metastases or not, the tumor deposit and budding were significantly higher in patients having overt metastases, compared with those not having them, in the Rad(-) group, whereas there was no significant difference in the Rad(+) group. CONCLUSIONS: Intramural spread and tumor deposit, budding, and micrometastasis were significantly decreased in the Rad(+) group, compared with the Rad(-) group. These results suggested effects of preoperative radiotherapy for sphincter preservation, as well as for decreasing the local recurrence rate. PMID- 15362754 TI - Endocrine tumors of the duodenum. A study of 55 cases relative to clinicopathological features and hormone content. AB - BACKGROUND/AIMS: Study of prognosis of duodenal endocrine tumors. METHODOLOGY: Retrospective study concerned 55 duodenal endocrine tumors discovered in biopsy or surgical specimens. Follow-up records available for 49 patients indicated that inconspicuous associated clinical manifestations were often found subsequently. Seven patients were classified as Zollinger-Ellison syndrome and seven as multiple endocrine neoplasia (6 MEN I and 1 MEN II). RESULTS: Tumors were small (mean 1.28cm) and located preferentially in the first and second part of the duodenum. Fifty-four were well-differentiated and one poorly differentiated. Immunochemistry revealed 30 G-cell tumors (54.6%), 15 D-cell (27.3%), two plurihormonal (EC cell and G cell), and one GRH-cell, whereas seven could not be classified. Fifteen patients died (five in relation to their disease). Twenty-one had metastases (liver, nodes, lung), eight of whom are still alive. CONCLUSIONS: Eighty-eight percent of duodenal endocrine tumors were gastrinomas, small plurifocal tumors and somatostatinomas preferentially located in the ampullar region and diagnosed because of hematemesis or icterus. Size is an important prognostic factor in determining whether surgery is required. The prognosis is better for D- and G-cell tumors than pancreatic endocrine tumors. Duodenal endocrine tumors in multiple endocrine neoplasia have a good prognosis, but can be associated with pancreatic plurihormonal tumors and metastases. PMID- 15362755 TI - Perforated duodenal ulcer at seven years after heart-renal transplantation: a case report. AB - We experienced a rare case of perforated duodenal ulcer that occurred at seven years after heart-kidney transplantation. This patient is reported here together with a discussion of the etiology, the selection of treatment, and perioperative management. The patient was a 46-year-old man who presented with precordial pain. In 1995, he had undergone simultaneous heart and kidney transplantation in the United States and had been on long-term immunosuppressive and corticosteroid therapy. His precordial pain started from May 24, 2002. He was examined at our hospital on May 27 and underwent emergency surgery with a diagnosis of upper gastrointestinal perforation. A 4-mm perforation was observed on the anterior wall of the duodenal bulb and panperitonitis was also present. Patch closure of the perforation was performed by pulling the omentum over the defect. Perioperative management consisted of his usual immunosuppressants together with antacid therapy. The postoperative course was good and he was discharged on hospital day 15. In this patient, the mechanism of perforation was assumed to involve sudden irritation combined with poor circulation in the duodenum and tissue ischemia, as well as a decrease of mucosal protective factors based on long-term corticosteroid therapy. Perforated duodenal ulcer is a rare problem after heart transplantation. Because the time that elapses after perforation is an important determinant of the prognosis, early diagnosis and appropriate surgical repair are essential. PMID- 15362756 TI - Responses of cytokines and coagulation-fibrinolytic states to surgical stress following esophagectomy. AB - BACKGROUND/AIMS: Surgery for esophageal cancer is one of the most invasive treatments in gastrointestinal surgery. The aim of this study was to analyze perioperative kinetics of coagulation-fibrinolytic states in comparison with cytokines in order to investigate biological responses to surgical stress in patients undergoing esophagectomy. METHODOLOGY: Serum or plasma samples of 20 patients with esophageal cancer who had undergone esophagectomy were collected during the perioperative period. Serial concentrations of cytokines (IL-6, IL-8, IL-10), PMN-E, and coagulation-fibrinolytic agents (TAT and PIC) were measured and analyzed. RESULTS: There were significant increases in concentrations of both inflammatory and anti-inflammatory cytokines during the operative and postoperative courses. TAT and PIC levels also increased significantly intra- and postoperatively. The TAT/PIC ratio was elevated simultaneously with a predominance of inflammatory cytokines over anti-inflammatory ones. CONCLUSIONS: In addition to the profile of cytokines, the TAT/PIC ratio seems to be a sensitive and useful indicator in evaluating biological responses to surgical stress. PMID- 15362757 TI - Combination treatment of transjugular retrograde obliteration and endoscopic embolization for portosystemic encephalopathy with esophageal varices. AB - The treatment of chronic portosystemic encephalopathy with esophageal varices has not yet been established. We were able to control a case of chronic portosystemic encephalopathy with esophageal varices using a combination treatment of transjugular retrograde obliteration and endoscopic embolization. A 57-year-old man came to our hospital in a confused, apathetic and tremulous state. The grade of encephalopathy was II. The plasma ammonia level was abnormally elevated to 119 microg/dL, and the ICGR15 was 59%. Endoscopic examination revealed nodular esophageal varices with cherry-red spots. There were no gastric varices. Ultrasonography and CT revealed liver cirrhosis with a splenorenal shunt. We first applied endoscopic embolization for the esophageal varices before transjugular retrograde obliteration. We injected 5% ethanolamine oleate with iopamidol retrogradely into the esophageal varices and their associated blood routes under fluoroscopy and obliterated the palisade vein, the cardiac venous plexus and left gastric vein. Transjugular retrograde obliteration was performed 14 days after endoscopic embolization. Retrograde shunt venography visualized the splenorenal shunt and communicating route to the retroperitoneal vein. There was no communicating route to the azygos vein. After obliteration of the communicating route to the retroperitoneal vein with absolute ethanol, 5% ethanolamine oleate with iopamidol was injected into the splenorenal shunt as far as the root of the posterior gastric vein. After transjugular retrograde obliteration, the encephalopathy improved to grade 0 even without the administration of lactulose and branched-chain amino acid. The plasma ammonia level and ICGR15 were reduced to 62 microg/dL and 26%. We conclude that combination treatment of transjugular retrograde obliteration and endoscopic embolization is a rational, effective and safe treatment for chronic portosystemic encephalopathy complicated with esophageal varices. PMID- 15362758 TI - Esophageal intramural spreading from an adenocarcinoma of the esophagogastric junction. AB - Although intramural spreading from gastric carcinoma to the esophageal wall is occasionally reported, longitudinal intramural lesion of the esophagus is very rare. We herein report the case of a patient found to have a carcinoma of the gastric cardia with intramural spreading to the esophagus approximately 7.0 cm in length. A 65-year-old man was admitted to our department suffering from a persistent midthoracic pain and mild dysphagia during the previous 3 months. Upper gastrointestinal studies revealed an oval submucosal tumor of the lower esophagus and a flare irregular lesion on the esophagogastric junction. An endoscopic ultrasonography showed the main tumor was in the submucosal layer and invaded beyond the muscularis propria. Histopathological examination of the resected specimen confirmed a poorly differentiated adenocarcinoma, 7.0 cm in length, which penetrated through the gastric wall, and invaded the submucosal layer of the esophagus. When only a few scattered carcinoma cells infiltrate only the mucosa or submucosa, it is difficult to diagnose the extent of esophageal invasion. In treating patients with gastric cancer with esophageal invasion, it is important to determine the safety of the proximal margin for esophageal resection. Histological examination using frozen sections obtained during surgery is essential for deciding the operative safety margin. PMID- 15362759 TI - Perioperative music therapy with a key-lighting keyboard system in elderly patients undergoing digestive tract surgery. AB - BACKGROUND/AIMS: It is important to minimize the perioperative mental dysfunction of elderly patients undergoing surgery and to avoid lowering their coping skills postoperatively. Music therapy for digestive tract surgery has yet not been explored. METHODOLOGY: We evaluated perioperative music therapy using a simple key-lighting keyboard system in 37 elderly patients who underwent digestive tract surgery (Group A) compared with 13 patients who were not applied music therapy (Group B). RESULTS: On the first day after surgery there were no general hemodynamic changes after music therapy. There were no significant changes in the Philadelphia Geriatric Center morale scale and the Yesavage depression scale between the day before surgery and 7 POD in both groups. The self-assessed visual analogue health scale and the number connection test worsened significantly from 58.9+/-14.6 and 159+/-47 to 42.3+/-14.6 and 199+/-51 (p<0.05), respectively, over this time in Group B, but it did not worsen significantly in Group A. The happiness score increased significantly from 3.9+/-1.1 to 4.6+/-1.2 (P<0.05) over this time in Group A, but it did not increase significantly in Group B. CONCLUSIONS: We conclude that the music therapy with a key-lighting keyboard system in elderly patients does not change postoperative hemodynamics and helps to maintain perioperative mental functioning. PMID- 15362760 TI - Is there a place for intraperitoneal onlay mesh repair (IPOM) of inguinal hernia among laparoscopic techniques? AB - BACKGROUND/AIMS: The Authors report their experience on laparoscopic hernioplasty using the intraperitoneal onlay mesh repair in 56 patients. METHODOLOGY: Thirty patients had a monolateral hernia, 9 of which were recurrent and 26 had a bilateral hernia, 6 of which were recurrent. Overall, a total of 90 hernias were treated. The hernia repair was performed by using "GORETEX Dual Mesh Plus biomaterial with holes" in the first 32 cases and the latest "Corduroy" type in the following 24 cases. The prostheses were fixed with titanium spiral tacks (Protack, Auto Suture, Tyco Healthcare). RESULTS: No intraoperative complications occurred and no conversion was necessary. Five minor postoperative complications (5.5%), 2 seromas and 3 transient paresthesias, were observed. Four patients (7.1%) needed analgesics after the first 24 hours. Mean hospital stay was 36 hours with a minimum of 24 and a maximum of 48. Mean resumption of normal activity was 8 days with return to work within two weeks. At an average 18 months follow-up, 3 recurrences were recorded (3.3%). CONCLUSIONS: The results of this study as well as the meta-analysis of the series presented in the literature, indicate that the intraperitoneal onlay mesh repair may be a feasible, safe and effective procedure in the treatment of recurrent and bilateral hernias or when a hernia repair is performed during other laparoscopic procedures. The intraperitoneal onlay mesh repair has in fact been shown to be faster and easier than the other more commonly performed laparoscopic hernioplasties (trans abdominal preperitoneal repair and total extra-peritoneal repair). These data may also suggest utilizing this technique in particular cases of primitive hernia such as very active young males or heavy-duty workers. However the limited series and the short follow-up ask for randomized prospective long-term studies to definitely ascertain the true incidence of recurrence and therefore the effectiveness of this attractive procedure. PMID- 15362761 TI - Systemic inflammatory response syndrome (SIRS) as a predictor of strangulated small bowel obstruction. AB - BACKGROUND/AIMS: Strangulated small bowel obstruction may often carry severe morbidity or mortality when a differential diagnosis from simple small bowel obstruction was made in the past. The aim of this study is to clarify preoperative indications that can be useful for predicting strangulation in acute small bowel obstruction, and to evaluate the efficacy of systemic inflammatory response syndrome as a predictor. METHODOLOGY: Seventy clinical cases were rated between strangulated (n=40) and simple (n=30) small bowel obstruction that were postoperatively identified and were analyzed using univariate and multivariate procedure. RESULTS: Age (p=0.0377), female gender (p=0.0121), white blood cells counts (p=0.0272), presence of abdominal tenderness (p=0.0074), presence of abdominal muscle guarding (p=0.0002), American Society of Anesthesiology score (p=0.0191), number of systemic inflammatory response syndrome factors (p=0.0001), and presence of systemic inflammatory response syndrome (p=0.0001) were evaluated for correlated predictive factors in the univariate analysis. The presence of systemic inflammatory response syndrome (odds ratio= 14.3, p=0.0151) and presence of abdominal muscle guarding (odds ratio=5.87, p=0.0158) were independent predictive factors in the multivariate analysis. CONCLUSIONS: These predictive factors were considered to be useful to predict strangulated small bowel obstruction, and to contribute to the improvement of outcomes in the management of acute small bowel obstruction. PMID- 15362762 TI - The results of caustic ingestions. AB - BACKGROUND/AIMS: The ingestion of caustic substances is a common condition, which may result in serious injuries of the upper gastrointestinal system. The purpose of this study is to assess the outcomes of patients who had a history of the ingestion of caustic substances. METHODOLOGY: Between January 1995 and May 2002, 53 patients with clinical and/or endoscopic signs of caustic agents injury admitted to the Emergency Surgery Service of University of Istanbul, Istanbul Medical School, were retrospectively evaluated. RESULTS: The group consisted of 29 females and 24 males with a mean age of 33 years (range 15 to 77). Caustic ingestion was accidental in 31 (59%) and suicidal in 22 (41%). Twenty-five (47%) patients had history of acid ingestions, 21 (40%) alkaline and 7 (12%) unknown agents. The findings of upper gastrointestinal tract endoscopy in 37 patients were as follows: i) fourteen patients had Grade I esophagitis, ii) fourteen patients had Grade II esophagitis; four with gastric mucosal burn, three with gastric and duodenal mucosal burn, five with pharyngeal mucosal burns, iii) two patients had gastric and duodenal mucosal burns, iv) four patients had gastric mucosal burn, vi) three patients had esophageal, gastric and duodenal mucosal burn. Eight patients (15%) with signs of acute abdomen underwent urgent surgery (six of these cases were acid ingestions and seven were suicidal ingestions); four of these patients with esophageal necrosis and gastric perforation, two with esophageal, gastric, duodenal and jejunal necrosis, one with local necrosis in the gastric fundus, and one had normal findings at laparotomy. The mean follow-up period was 11.6 months (range 1 to 42 months). During the follow-up one patient presented with pyloric stenosis, which required surgery. The mortality rate was 11%. CONCLUSIONS: Suicidal caustic ingestion and acid ingestion seem to cause more severe injuries. Early admission to the hospital with clinical and endoscopic evaluation and early surgery when required may reduce morbidity and mortality. PMID- 15362763 TI - Total intraoperative enteroscopy using a colonoscope for detecting the bleeding point. AB - We report a case of bleeding in the small intestine of a 59-year-old man that was successfully diagnosed and treated by total intraoperative enteroscopy. The patient was admitted to our hospital because of gastrointestinal bleeding. Gastroscopy, colonoscopy and mesenteric arteriography could not identify the bleeding point. Technetium-99m-labeled red blood cell scintigraphy suggested bleeding from the small intestine but could not reveal the precise lesion. As the patient presented melena and went into a state of shock, we performed emergency laparotomy with total intraoperative enteroscopy using a colonoscope. A small ulceration was found at the jejunum, and subsequently partial resection of the jejunum was performed. The lesion was diagnosed histopathologically as angiodysplasia. The patient recovered uneventfully and was discharged on the 24th postoperative day. Bleeding in the small intestine is rare and difficult to diagnose but it sometimes induces a severe condition. Total intraoperative enteroscopy using a colonoscope is effective for detecting the bleeding point in the small intestine and can be performed at most centers without special fiberscopes and techniques. PMID- 15362764 TI - Effect of sodium nitroprusside on ischemia-reperfusion injuries of the rat liver. AB - BACKGROUND/AIMS: The role of NO (nitric oxide) in liver ischemia-reperfusion injury remains controversial, therefore the effect of sodium nitroprusside (SNP), as NO donor on ischemia-reperfusion injuries of the rat liver was studied. METHODOLOGY: Warm ischemia (68% of total liver) of the rat liver was induced for 30 min, followed by reperfusion for 60 min. A dose of 1 or 10 microg/kg/min SNP were continuously administered during the reperfusion period. We performed the following assessments: (i) hepatic microcirculation, (ii) nitrosyl-hemoglobin (Hb NO, a product of interactions between hemoglobin and NO) levels, (iii) NO2-/NO3- (end products of NO oxidation) levels, and (iv) the serum lactate-dehydrogenase (LDH) levels. RESULTS: Hepatic microcirculation at 5 and 15 min after start of SNP infusion was significantly improved, compared with that in the control group. NO2-/NO3- and Hb-NO levels in both SNP groups were gradually elevated after reperfusion. Serum LDH levels at 60 min after start of reperfusion in both SNP groups were significantly lower than in the control group. CONCLUSIONS: In the early phase of the reperfusion period, SNP injected increased hepatic microcirculation and reduced hepatocyte damage, suggesting that the administration of SNP may help present ischemia-reperfusion injuries. PMID- 15362765 TI - The role of duplex-doppler ultrasonography in the diagnosis of renal dysfunction and hepatorenal syndrome in patients with liver cirrhosis. AB - BACKGROUND/AIMS: Kidney dysfunction commonly develops in patients with liver cirrhosis. Renal failure develops due to renal vascular constriction and can be present weeks or even months before clinical signs or increased levels of blood urea nitrogen or serum creatinine concentrations become detectible. The objective of this study was to analyze the value of renal interlobar arterial resistance index, measured by duplex-Doppler ultrasonography for detecting early impairment of renal function in patients with liver cirrhosis and its possible use in detecting a subgroup of patients with a higher risk of developing hepatorenal syndrome. METHODOLOGY: The patients were divided into three groups: patients with liver cirrhosis and normal renal function (n=31), patients with liver cirrhosis and renal failure but without hepatorenal syndrome criteria (n=9), and patients with hepatorenal syndrome (n=6). The interlobar arterial resistance index was estimated with duplex Doppler ultrasonography, and liver and renal function tests were measured in all patients. RESULTS: The average value of interlobar arterial resistance index in patients with hepatorenal syndrome (0.74+/-0.01) was statistically significantly higher than interlobar arterial resistance index values measured in liver cirrhosis patients without the signs of azotemia (0.65+/ 0.03) or in those with liver cirrhosis and kidney dysfunction, but without hepatorenal syndrome (0.67+/-0.01). In all patients with hepatorenal syndrome the value of interlobar arterial resistance index was over 0.70. In the group of patients with liver cirrhosis and kidney dysfunction, but without hepatorenal syndrome, interlobar arterial resistance index was below 0.70 in seven, whereas in the remaining two interlobar arterial resistance index was above 0.70. In those patients renal dysfunction displayed a progressive form and hepatorenal syndrome developed. CONCLUSIONS: Duplex-Doppler ultrasound of intralobar arteries is a simple, effective and non-invasive method which enables the early detection of renal hemodynamic disturbances in patients with liver cirrhosis even before renal dysfunction becomes clinically evident. It also makes possible the identification of a subgroup of patients with liver cirrhosis who are at higher risks for developing hepatorenal syndrome. PMID- 15362766 TI - Synergistic effect of cold and warm ischemia time on postoperative graft outcome in human liver transplantation. AB - BACKGROUND/AIMS: Prolonged cold ischemia time (CIT) during graft preservation and warm ischemia time (WIT) defined as a rewarming time have been reported to cause postoperative graft dysfunction after orthotopic liver transplantation (OLT). However, a synergistic effect of both CIT and WIT on patients of graft survival has not been confirmed. The aim of this study was to determine whether simultaneously prolonged CIT and WIT was associated with early graft outcome after clinical OLT. METHODOLOGY: Between May 1997 and July 1998, 186 consecutive OLT cases were divided into 4 groups as follows: group A, CIT < or =12 hrs and WIT < or =45 min; group B, CIT >12 hrs and WIT < or =45 min; group C, CIT < or =12 hrs and WIT >45 min; and group D, CIT > 12 hrs and WIT >45 min. Liver graft survival within 90 days of OLT and early postoperative graft function were analyzed. RESULTS: The graft loss rates were 5.4% in group A, 9.8% in group B, 11.1% in group C, and 42.9% in group D. The mean highest aspartate aminotransferase (AST) values after OLT in group D (3352.3+/-569.4 U/L) was significantly higher than those in groups A (1411.7+/-169.2 U/L) and B (1931.3+/ 362.6 U/L). CONCLUSIONS: The simultaneously prolonged cold and warm ischemia time significantly caused hepatic allograft injury and failure, suggesting some synergistic effects of CIT and WIT on postoperative graft function. PMID- 15362767 TI - Inhibition of activated blood platelets by interferon alpha 2b in chronic hepatitis C. AB - BACKGROUND/AIMS: Interferon alpha used in treatment of chronic hepatitis C significantly influences the blood platelets. The role of platelets in initiating and developing pathological processes in hepatic diseases is still barely known. We studied the effects of interferon alpha 2b (IFN alpha2b) on blood platelets in chronic hepatitis C. METHODOLOGY: The studies were conducted in 16 patients who underwent IFN alpha2b treatment 3 times a week at 6MU. The examination was carried out before and on the 14th day of the treatment of IFN alpha2b. Morphological parameters of blood platelets were determined by hematological methods and flow cytometry. Expression of receptors on blood platelet surfaces (CD41, CD42a, CD62P) and thrombopoietin, platelet-derived growth factor, soluble form sP-selectin, IL-6, and tumor necrosis factor alpha were also determined. RESULTS: The use of IFN alpha2b in patients with chronic hepatitis C significantly effects blood platelets morphology by causing the decrease in their number, the change in population size, and the increase in large platelet count. Interferon decreases P-selectin expression on platelets, sP-selectin and platelet derived growth factor concentration in plasma. During interferon therapy we noted increase concentration of thrombopoietin, tumor necrosis factor alpha, IL-6 in chronic hepatitis C. CONCLUSIONS: IFN alpha2b stabilizes activated platelets and probably decreases their participation in inflammatory and fibrotic processes in the liver. PMID- 15362768 TI - Hepatic resection induces a shift in the Th 1/2 balance toward Th 2 and produces hypermetabolic and hyperhemodynamic states. AB - BACKGROUND/AIMS: Recent studies have shown that the type 1/2 T-helper (Th 1/2) cell balance is shifted toward a Th 2-type immune response not only by malignancy but also by surgical stress. The present study evaluated surgical stress as a result of liver surgery in comparison with that of other major abdominal surgeries. Immune, metabolic and circulatory responses to surgery were measured for determination of surgical stress. METHODOLOGY: Eighty-five patients who underwent abdominal surgeries were divided into three groups: hepatic resection (n=17), gastric resection (n=38), colorectal resection (n= 30). Blood sampling was performed before surgery, and on postoperative days (POD) 2 and 14. The Th 1/2 helper T cell balance was determined by flow cytometric analysis of interferon-gamma and interleukin-4 expression. Energy expenditure was measured by indirect calorimetry, and hemodynamics was studied using pulse dye densitometry until POD 14. RESULTS: Following surgery Th 1/2 ratios decreased significantly. Additionally, the Th 1/2 balance in patients with hepatic resection on POD 2 was significantly lower than that of patients with other major surgeries. However, on POD 14 there were no significant differences among the three groups. Resting energy expenditure and cardiac index on postoperative days 1 and 3 in patients with hepatic resection increased significantly above levels in the other surgical groups. Conversely, blood volume in the hepatic resection patients was significantly lower than that of other patients until POD 3. Ten patients who developed postoperative complications had significantly lower Th 1/2 ratios and more hypermetabolism. CONCLUSIONS: This study reveals that hepatic resection induces a more marked shift toward a Th 2 helper T cell response and significantly more hypermetabolism than other major surgeries. A distinct pattern of Th 1/2 ratio changes during the early phase of the postoperative course in hepatic resection may be related to changes in metabolism and circulation. Therefore, determination of Th 1/2 balance may be of help in evaluating different surgical procedures, and management of energy intake and circulatory management may be cautiously determined based on the shift in Th 1/2 balance. PMID- 15362769 TI - Long-term survival after pulmonary metastatectomy of hepatocellular carcinoma; treatment outcome or natural history? AB - BACKGROUND/AIMS: To evaluate the role of pulmonary metastatectomy in hepatocellular carcinoma. METHODOLOGY: Four hepatocellular carcinoma patients underwent pulmonary metastatectomy. Patients were considered candidates for pulmonary metastatectomy when they met the following criteria; 1) intrahepatic hepatocellular carcinoma was under control or anticipated to be under control in the near future; 2) the metastatic lesions appeared to be amenable to complete resection; 3) there was no metastasis at another site. RESULTS: The median duration of follow-up was 4.1 (range: 2.4 to 6.0) years. During this period, 1 patient died of pulmonary recurrence 29 months after metastatectomy. The remaining 3 were alive at the time of writing; one for 43 months after metastatectomy without recurrence; another for 72 months with intrahepatic recurrence; and the third for 54 months with extrahepatic recurrence. CONCLUSIONS: Long-term survival was achieved after pulmonary metastatectomy in highly selected hepatocellular carcinoma patients. However, "highly selected patients" might imply that such patients belong to a special subgroup composed of cases with an intrinsically slow growing tumor biology. Therefore, it is an issue of interest as to whether the long-term survival acquired by these cases results from the pulmonary metastatectomy, or whether it is a reflection of a fate dictated by intrinsic tumor biology. PMID- 15362770 TI - The clinical significance of monitoring alkaline phosphatase level to estimate postoperative liver failure after hepatectomy. AB - BACKGROUND/AIMS: To predict the occurrence of postoperative liver failure after hepatectomy, the clinical significance of monitoring alkaline phosphatase (ALP) has been studied, and the relationship between the change of total bilirubin and the level of ALP or gamma-glutamyl transpeptidase (gamma-GTP) after hepatectomy was evaluated. METHODOLOGY: The 163 patients, who underwent hepatectomy at our institute for the past 12 years, were divided into three groups according to the postoperative events. HF consisted of 5 patients with liver failure, HB showed the postoperative high bilirubinemia (over 5mg/dL) in 13 cases and GP were 145 cases without any postoperative problems. RESULTS: 1. The postoperative highest level of total bilirubin (T-Bil) correlated with the decreasing rate of ALP, prothrombin time (PT), total cholesterol (T-CHO) or gamma-GTP and total blood loss (p<0.01). 2. The level of ALP decreased after hepatectomy significantly and the decreasing rate was serious in HB and HF (p<0.05). 3. The recovering time to preoperative level of ALP was clearly shorter in GP than in HB and HF. 4. The level of ALP and gamma-GTP at the point where the level of T-Bil increased over 5mg/dL, was useful to distinguish between HF and HB. 5. The good correlation between postoperative level of ALP and gamma-GTP was noted. The decreasing levels of ALP and gamma-GTP were found to be critical below 80% and 55% after bisegmentectomy. Furthermore, the recovered levels of ALP and gamma-GTP were important to distinguish between HF and HB. CONCLUSIONS: Monitoring the ALP level was indicated to be useful to estimate the postoperative course of bilirubin. PMID- 15362771 TI - Protective effect of methylprednisolone and of intermittent hepatic pedicle clamping during liver vascular inflow occlusion in the rat. AB - BACKGROUND/AIMS: One of the main causes of postoperative morbidity and mortality following major hepatic resection is hepatic ischemia deliberately designed to reduce intraoperative hemorrhage. This study assessed the effects of intermittent or continuous hepatic ischemia and reperfusion with or without methylprednisolone pretreatment in the rat. METHODOLOGY: One hundred and eighty rats were divided into 3 groups undergoing hepatic ischemia of 60, 90, and 120 minutes total duration. Each group of rats were subdivided to receive either a continuous Pringle maneuver, or 30 min or 15 min of intermittent liver pedicle clamping. Ten minutes before ischemia induction, 10 rats from each group were pretreated with intravenous 3 mg/100 g bw methylprednisolone. RESULTS: With continuous hepatic pedicle clamping the rat survival rates inversely correlated with the duration of ischemia (survival: 70%, 40%, and 20% with ischemia of 60, 90, and 120 min). Survival rates at 15-min and 30-min intermittent ischemia groups were significantly higher than in the continuous clamping group (p<0.05). Methylprednisolone pretreatment did not significantly increase survival but resulted as a significant reduction in liver enzyme release (AST, ALT), at 90 min (p<0.05) and at 120 min (p<0.05) in the continuously clamped groups. When ischemia lasted 120 min, methylprednisolone pretreatment was associated with higher preservation of ATP liver content (p<0.05). CONCLUSIONS: This study confirms that intermittent hepatic pedicle clamping significantly improves survival in rats undergoing hepatic vascular inflow occlusion with a decrease in transaminase release and greater maintenance of intrahepatic ATP after prolonged total ischemia when animals were pretreated with methylprednisolone. PMID- 15362772 TI - Combination transcatheter hepatic arterial chemoembolization with thymosin alpha1 on recurrence prevention of hepatocellular carcinoma. AB - BACKGROUND/AIMS: To observe recurrence prevention on hepatocellular carcinoma after hepatectomy by using TACE (transcatheter hepatic arterial chemoembolization) with thymosin alpha1 postoperatively. METHODOLOGY: From January 2000 to December 2002, 57 patients with hepatocellular carcinoma were randomly divided into three groups: Group A (n=18) received hepatectomy plus TACE and thymosin alpha1 postoperatively, group B (n=23) received hepatectomy plus TACE postoperatively and group C (n=16) received hepatectomy only. The recurrent rate, recurrent time and median survival period for the three groups were observed and measured. RESULTS: For group A, B and C, one-year recurrent rates were 83.3%, 87.0% and 87.5% (p=0.926), respectively. The recurrent time were 7.0, 5.0 and 4.0 months (p=0.039), respectively, and the median survival were 10.0, 7.0 and 8.0 months (p=0.002), respectively. CONCLUSIONS: Comprehensive therapy combining TACE plus Talpha1 postoperatively could not decrease the recurrent rate, but it might delay the recurrent time and prolong survival periods for hepatocellular carcinoma patients after hepatectomy. PMID- 15362773 TI - Intraoperative three-dimensional visualization of liver vasculature by ultrasonography. AB - BACKGROUND/AIMS: To understand the three-dimensional relationship between the liver vasculature and tumor by intraoperative sonography, some training is inevitable. Three-dimensional ultrasound has been evaluated in various fields, but not yet in intraoperative sonography. METHODOLOGY: Extracorporeal 3-D ultrasonography was performed in 32 patients. Of these, 20 underwent intraoperative 3-D ultrasonography. Using these images, we evaluated whether or not the portal vein, hepatic vein and its branches were discernible. RESULTS: Satisfactory images were obtained in all 32 patients by extracorporeal methods and in 15 by intraoperative 3-D ultrasonography. The number of visualized veins, including the right portal vein, its anterior branch, posterior branch, anterior superior branch, anterior inferior branch and right hepatic vein, was 32, 32, 23, 28, 22, and 30 by extracorporeal 3-D ultrasonography, and 15, 14, 10, 8, 5, and 12 by intraoperative 3-D ultrasonography. The relationships between the intrahepatic artery, portal vein, and hepatic vein were more clearly visualized by intraoperative 3-D sonography. CONCLUSIONS: Intraoperative 3-D ultrasonography clearly showed small branches of the liver vasculature and their 3-D relation, which may be helpful for liver surgery. However, this method requires further improvement. PMID- 15362774 TI - Serum C3 complement concentrations correlate with liver function in patients with liver cirrhosis. AB - BACKGROUND/AIMS: The aim of this study was to investigate whether C3 and C4 serum complement concentrations have prognostic relevance for patients with liver cirrhosis. METHODOLOGY: Serum complement concentrations of C3 and C4 were measured in 69 patients with liver cirrhosis and correlated with the Child-Pugh score. RESULTS: C3 concentrations were 1.06+/-0.21 g/L in patients with Child Pugh A liver cirrhosis and significantly lower in Child-Pugh B (0.78+/-0.24 g/L) and even lower Child-Pugh C (0.49+/-0.14 g/L) (p=0.006 B vs. A, p<0.001 C vs. B). Patients with consecutive hepatorenal syndrome (HRS) had the lowest C3 concentrations (0.44+/-0.05 g/L (Child-Pugh C +HRS) vs. 0.54+/-0.06g/dL (Child Pugh C -HRS); p<0.05). C4 concentrations were 0.21+/-0.08 in Child-Pugh A and significantly lower in Child-Pugh B (0.11+/-0.04) and Child-Pugh C (0.09+/-0.04) patients (p<0.001). There was a negative correlation between C3 (r = -0.81, p<0.001) and C4 (r = -0.51, p<0.05) concentrations and the Child-Pugh score. CONCLUSIONS: Serum complement concentrations of C3 and C4 correlate negatively with the Child-Pugh score in patients with liver cirrhosis. C3 concentrations are lower in those Child-Pugh C cirrhosis patients with consecutive development of HRS. PMID- 15362775 TI - Clinical significance of plasma D-dimer levels and serum VEGF levels in patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: Angiogenesis and coagulation system activation are associated with tumor growth and metastasis. Vascular endothelial growth factor (VEGF) has been reported to play a major role in tumor angiogenesis. The elevation of plasma D-dimer level indicates the activation of coagulation and fibrinolysis. The purpose of this study was to: (a) evaluate the correlation between serum VEGF and plasma D-dimer level; (b) analyze the clinical features that might affect the VEGF and D-dimer levels in patients with hepatocellular carcinoma. METHODOLOGY: Twenty patients with hepatocellular carcinoma were included prior to treatment. Serum VEGF levels were measured by enzyme-linked immunosorbent assay. Plasma D dimer levels were measured by quantitative latex microparticle enhanced turbidimetric immunoassay. RESULTS: The presence of a high plasma D-dimer level was found to be correlated with the presence of central necrosis, higher Child's grade, advanced TNM stage, and the presence of portal vein thrombosis when plasma D-dimer levels were compared between different clinicopathologic groups. Tumors larger than 2 cm in diameter had higher median serum VEGF levels than tumors less than 2cm in diameter. No correlation was found between plasma D-dimer level and serum VEGF level in hepatocellular carcinoma patients (r=0.126, p=0.598). CONCLUSIONS: No correlation was found between the plasma D-dimer level and the serum VEGF level in hepatocellular carcinoma patients. The plasma D-dimer level appeared to reflect the tumor stage and vascular invasion of hepatocellular carcinoma. Serum VEGF level in hepatocellular carcinoma patients showed a positive correlation with tumor size. PMID- 15362776 TI - Hepatic resection through an anterior approach employing a modified liver hanging maneuver in patients with a massive liver tumor severely oppressing the inferior vena cava. AB - For a large hepatic neoplasm existing in the right hepatic lobe, hepatic resection using an anterior approach is required. We have reported an operative procedure for hepatic transection using absorbable polyglycolic acid tape. In patients with suspected tumor invasion of the inferior vena cava, on the other hand, considering the range of the residual tumor while sparing the inferior vena cava as much as possible, combined resection and reconstruction of the inferior vena cava is conducted only if operative curativity is expected. We conducted hepatic transection while maintaining the blood flow of the residual liver by applying the liver hanging maneuver method of Belghiti et al. and polyglycolic acid tape in patients with giant liver tumors of the right hepatic lobe compressing the hepatic inferior vena cava. Strong angled dissecting forceps were inserted into the ventral side of the inferior vena cava from the caudal side, and the tip was induced between hepatic veins. Two strips of polyglycolic acid tape were pinched with forceps and strongly ligated on the right and left sides of the cutoff line. Subsequently, hepatic transection was conducted using electrocautery spray coagulation and CUSA without blocking the inflow blood of the residual liver, and the right hepatic lobe was extirpated. This procedure has already been performed in 5 patients suspected of inferior vena cava invasion, and the inferior vena cava was able to be preserved in all the patients. PMID- 15362777 TI - Hanging maneuver in left hepatectomy. AB - The liver hanging maneuver in right hepatectomy introduced by Belghiti in 2001, is a safe and effective method for right hepatectomy in cases of large hepatoma and living donor liver transplantation. In this article, we first introduce the hanging maneuver in extended left hepatectomy (left hepatectomy including middle hepatic vein). Extended left hepatectomy is a more difficult procedure than right hepatectomy because in the deeper part of the transection, the plane sharply turns to the left above the caudate lobe. Using the hanging maneuver, the horizontal transection plane becomes vertical, and the extended left hepatectomy can be performed easier and safer. This technique can be applied effectively in all kinds of left-sided hepatectomies. PMID- 15362778 TI - Surgical approach to liver metastasis with hepatic hilar invasion. AB - We present a surgical procedure for liver metastases with hepatic hilar invasion, which was originally developed for hilar bile duct cancer. A 64-year-old man with 8 metastatic nodules from colon cancer and hepatic hilar invasion received this technique, and lived for more than 5 years without any restriction in his life. The authors recommend this procedure for those patients in order to preserve their quality of life. PMID- 15362779 TI - Clinical experience of transjugular intrahepatic portosystemic shunt (TIPS) and its effects on systemic hemodynamics. AB - We performed TIPS (transjugular intrahepatic portosystemic shunt) in patients with intractable esophageal varices accompanied by repeated hematemesis or with refractory ascites for the purpose of portal venous decompression, and successfully obtained complete elimination of esophageal varices or a marked decrease in ascites. While TIPS caused no particular variations in mean blood pressure or heart rate, cardiac output increased markedly on the 2nd and 3rd postoperative days before declining on the 5th postoperative day. Along with this, right atrial pressure, pulmonary arterial pressure and pulmonary capillary wedge pressure also increased transiently. TIPS has the potential to become an established effectual therapy for intractable esophageal varices and refractory ascites. However, careful attention should be paid to its hemodynamic effects, including the occurrence of cardiac failure. PMID- 15362780 TI - Lethal hypercytokinemia following hepatic resection under pringle maneuver: a case report. AB - Clinical implications of acute reactant cytokines remain to be clarified in ischemia/reperfusion injury of humans. We report a lethal case of hypercytokinemia following continuous Pringle maneuver. A 36-year-old man with intrahepatic duct stones underwent left lobectomy under continuous hepatic inflow occlusion for 70 minutes. The postoperative course was stormy with rapid deterioration of liver functions, resulting in death due to multiorgan dysfunction on the 4th postoperative day. Analysis of cytokines demonstrated marked elevation of plasma acute inflammatory cytokines level (Interleukin-6 and 8) during surgery and immediate postoperative day. Our experience suggests that excessive production of inflammatory cytokines was detrimentally associated with multiorgan dysfunction including liver. The strategies against such hypercytokinemia should be considered when performing liver resection particularly under continuous Pringle maneuver. PMID- 15362781 TI - Successful application of highly purified natural interferon alpha (multiferon) in a chronic hepatitis C patient resistant to preceding treatment approaches. AB - A currently 65-year-old patient with histologically proven chronic hepatitis C and chronic hepatitis B (seroconversion in 1990) and additional compensated cirrhosis of the liver (Child A) achieved sustained complete biochemical and viral response following 5 and 14 months respectively of therapy with highly purified natural leukocyte interferon-alpha (3 x 3 MU weekly, nIFN-alpha, Multiferon). Prior to this treatment, various other therapy approaches including recombinant interferon-alpha2b (rIFN-alpha2b) or a combination of natural interferon-beta (nIFN-beta) and interferon-gamma (rIFN-gamma) had been carried out. Unfortunately, these had been unsuccessful. After a total treatment period of 76 months with nIFN-alpha and a subsequent follow-up period of 30 months, no relapse of chronic hepatitis C occurred. The patient's tolerance of the treatment was excellent and no substantial drug-related adverse events were observed. nIFN alpha, which - unlike the recombinant IFN-alpha2 preparations - is a mixture of various physiologically expressed IFN-alpha subtypes, could possibly be an alternative in the treatment of difficult-to-treat patients with chronic hepatitis C. PMID- 15362782 TI - Invasive carcinoma derived from intraductal papillary mucinous carcinoma of the pancreas. AB - BACKGROUND/AIMS: Most patients with intraductal papillary mucinous tumors of the pancreas have a favorable prognosis after surgical treatment. However, recurrent disease frequently occurs in patients with invasive carcinoma derived from intraductal papillary mucinous carcinoma. The objective of this study was to clarify the clinicopathological features of invasive carcinoma derived from intraductal papillary mucinous carcinoma. METHODOLOGY: We performed a retrospective review of the 29 patients with intraductal papillary mucinous tumor including 10 patients with invasive carcinoma who underwent pancreatic resection between June 1995 and December 2001 at the National Cancer Center Hospital East. RESULTS: Of 10 patients with invasive carcinoma derived from intraductal papillary mucinous carcinoma, 7 patients had lymph node involvement and 8 patients had retroperitoneal invasion. The overall 1-, 2-, 4-year actuarial survival rate for invasive carcinoma derived from intraductal papillary mucinous carcinoma was 39%, 26%, 13%. Recurrence occurred as liver metastasis in 3 patients, carcinomatous peritonitis in 3, local recurrence in 3, and lung metastasis in 1. All patients with adenoma, non-invasive carcinoma, and minimally invasive carcinoma are alive without recurrent disease after pancreatic resection. CONCLUSIONS: Patients with invasive carcinoma derived from intraductal papillary mucinous carcinoma had a worse prognosis. Margin-negative pancreatic resection is essential for treating this disease. PMID- 15362783 TI - Risk factors of pancreatic fistula following pancreaticoduodenectomy for periampullary cancer. AB - BACKGROUND/AIMS: Pancreatic fistula remains one of the most common complications of pancreaticoduodenectomy. This study was carried out to identify relevant risk factors for the development of pancreatic fistula and to evaluate whether prophylactic administration of octreotide decreases their incidence. METHODOLOGY: Between March 1985 and December 2001, 129 consecutive patients who underwent elective pancreaticoduodenectomy for pancreatic or ampullary carcinomas were prospectively evaluated. Pancreatic fistula was defined as drainage of at least 10 mL of amylase-rich fluid detected in abdominal drains after the 3rd postoperative day. RESULTS: The overall postoperative complication rate was 36.4% (n=47) and pancreatic fistula was found in 12.4% (n=16) of patients. No differences were found between patients receiving octreotide and the non octreotide group regarding the incidence of postoperative complications, including pancreatic fistula. Univariate and multivariate analysis of factors predicting the development of pancreatic fistula showed that only soft pancreatic texture (p=0.0002) and ligation of the pancreatic remnant (p=0.029) were significantly associated with fistula formation. CONCLUSIONS: The only surgeon dependent factor which may reduce the incidence of stump-related complications is anastomosis of the pancreatic remnant with the alimentary tract. Carried out single-institution analysis showed octreotide to be ineffective in the prevention of pancreatic fistula following pancreaticoduodenectomy. PMID- 15362784 TI - Intraductal papillary-mucinous tumors of the pancreas. AB - BACKGROUND/AIMS: To elucidate the risk of malignancy and the morphological alterations associated with malignancy. METHODOLOGY: Thirty cases of intraductal papillary-mucinous tumors and 5 papillary-mucinous carcinomas (invasive intraductal papillary-mucinous tumors) of the pancreas were clinicopathologically and histopathologically analyzed. RESULTS: The invasive carcinoma developed on the basis of severe dysplasia-carcinoma in situ changes and never from mild or moderate dysplasia changes. However, tumor cell projections of intraductal papillary-mucinous tumors encroached into the duct wall and/or the stroma introduced just beneath the epithelium and "intraductal" tumor cells sometimes came in direct contact with the "extraductal" connective tissues even in adenomas. The frankly invasive adenocarcinoma components of invasive intraductal papillary-mucinous tumors were characterized by the lack or poor formation of their own basement membrane and were usually surrounded by the extensive collagenous proliferation, desmoplastic reaction. Such stromal alterations never developed around the "extraductal" components of non-invasive intraductal papillary-mucinous tumors. CONCLUSIONS: The risk of malignancy for an individual intraductal papillary-mucinous tumor was increased with the degree of cellular and/or structural atypia. The desmoplastic reaction with poor formation of the basement membrane is the sine qua non of the "true invasion". PMID- 15362785 TI - A survival predictive model in patients undergoing radical resection of ampullary adenocarcinoma. AB - BACKGROUND/AIMS: Radical resection with either pancreaticoduodenectomy or pylorus preserving pancreaticoduodenectomy is considered to be the standard treatment for most ampullary carcinomas, but the prognostic predictive model has not yet been developed. METHODOLOGY: The pretreatment, treatment, and follow-up variables of data of 47 patients undergoing radical resection for the ampullary carcinoma were analyzed to determine the favorable prognostic variables. Employing the Kaplan Meier method, the cumulative survival rates of the ampullary carcinoma were calculated. By Cox regression model, a stepwise multivariate analysis was performed to analyze the contributing factors of the survival rate, and a predictive survival equation was obtained. RESULTS: With the results of the univariate analysis, the variables significantly associated with favorable prognosis were younger age (<63 years), TNM stage (stage I or II or III), and the degree of tumor differentiation (well or moderately differentiated). When the above three variables were examined as covariates by Cox regression in multivariate analysis, the TNM stage and the degree of tumor differentiation were independently correlated with the survival. A predictive survival equation obtained with the beta-coefficients of the above three variables was as follows: S (t) = [So (t)] P, P = exp (0.0234 x age - 1.8744 x tumor differentiation + 1.1576 x TNM stage) CONCLUSIONS: This predictive survival equation can predict the survival and the favorable outcome of patients treated with radical resection of ampullary carcinoma. PMID- 15362786 TI - Pancreatic liver metastases after curative resection combined with intraoperative radiation for pancreatic cancer. AB - BACKGROUND/AIMS: A high proportion of patients even after curative resection for pancreatic cancer suffer from hepatic metastases. The aim of this study was to identify clinicopathological predictors of liver metastases after surgery, retrospectively. METHODOLOGY: Forty-one patients underwent extended radical pancreatectomy combined with intraoperative radiotherapy, which is one of the best local control methods for ductal cell carcinoma of the pancreas. Of the 41 patients, twenty-one patients regarded as being in a cancer free state after this combined therapy were studied to analyze clinicopathological predictors of hepatic metastases. Odds ratios and their 95% confidence intervals were calculated from data using logistic regression analysis. Statistical difference was considered significant at p<0.05. RESULTS: Liver metastases after curative resection occurred in 11 patients. Preoperative biliary drainage, jaundice, elevated preoperative serum tumor-associated carbohydrate antigens levels, microscopic distal bile duct invasion, duodenal wall invasion, extrapancreatic nerve plexuses invasion were factors influencing postoperative liver metastases. CONCLUSIONS: We found clinicopathological predictors of postoperative liver metastases. Patients with these factors require consideration in careful follow up and perioperative adjuvant therapy for prevention of postoperative liver metastases. PMID- 15362787 TI - Inferior head resection of the pancreas and cyst resection for choledochal cyst with chronic calcifying pancreatitis. AB - We report inferior head resection of the pancreas and cyst resection for congenital choledochal cyst with an anomalous arrangement of pancreaticobiliary duct and chronic calcifying pancreatitis. A 42-year-old man was admitted to the National Cancer Center Hospital East complaining of back pain. Contrast-enhanced computed tomography showed marked dilatation of the bile duct and multiple pancreatic stones in the main pancreatic duct. Endoscopic retrograde cholangiopancreatography demonstrated pancreatic stones in the dilated main pancreatic duct. The patient underwent cyst excision, inferior head resection of the pancreas, hepaticojejunostomy and lateral pancreaticojejunostomy. The postoperative course was uneventful. This procedure relieved the back pain. Choledochal cyst with anomalous arrangement of the pancreaticobiliary duct is frequently associated with acute pancreatitis. Inferior head resection of the pancreas removed the common channel which could be the cause of relapsing pancreatitis. Thus, inferior head resection can play a role in the management of choledochal cyst with chronic pancreatitis. PMID- 15362788 TI - Combination treatment of partial splenic embolization, endoscopic embolization and transjugular retrograde obliteration for complicated gastroesophageal varices. AB - The treatment of complicated gastroesophageal varices with a gastrorenal shunt and portal hypertensive gastropathy has not yet been established. We were able to control a case of complicated large gastroesophageal varices with gastrorenal shunt and portal hypertensive gastropathy using a combination treatment of partial splenic embolization, endoscopic embolization and transjugular retrograde obliteration. We first applied partial splenic embolization to reduce the hyperdynamic cycle of portal hypertension and to treat thrombocytopenia. We secondarily applied endoscopic embolization for the esophageal varices. Transjugular retrograde obliteration was performed for the gastric varices 14 days after endoscopic embolization. The wedged hepatic venous pressure had not changed after transjugular retrograde obliteration. After the combination treatment, the gastroesophageal varices were successfully obliterated, but portal hypertensive gastropathy did not worsen. The platelets count, arterial ketone body ratio and ICGR15 were improved. Partial splenic embolization was useful to protect side effects of endoscopic embolization and transjugular retrograde obliteration. We conclude that the combination treatment of partial splenic embolization, endoscopic embolization and transjugular retrograde obliteration is a rational, effective and safe treatment for complicated gastroesophageal varices with gastrorenal shunt and portal hypertensive gastropathy. PMID- 15362789 TI - Iron deficiency anemia after subtotal gastrectomy for gastric cancer. AB - BACKGROUND/AIMS: Sideropenic anemia after a gastrectomy is a frequent complication. The aim of the present study was to evaluate the role of different factors, such as sex, age, atrophic chronic gastritis, Helicobacter pylori infection and iron malabsorption, in iron deficiency after surgery for gastric cancer. METHODOLOGY: Thirty-seven patients who underwent subtotal gastrectomy for carcinoma of the stomach were prospectively studied following a specific three year protocol. Iron deficiency was evaluated by hemochromocytometric analysis and serum iron-ferritin level assays. RESULTS: Of the different variables analyzed, atrophic chronic gastritis was associated with a lower mean serum iron level, in particular two years after surgery (65mg/dL vs. 103 mg/dL in subjects without gastritis, P<0.01); a correlation between Helicobacter pylori infection of the gastric stump and lower mean serum ferritin level was also found (25+/-6.3 mg/dL vs. 53+/-0.4 mg/dL, P<0.05). On the contrary, no association was observed with the other factors that were evaluated. CONCLUSIONS: Among the factors involved in iron deficiency after gastrectomy for cancer of the stomach, atrophic gastritis seems to be the most important, although Helicobacter pylori infection of the gastric stump also seems to play an important role. PMID- 15362790 TI - Effect of H. pylori on COX-2 expression in gastric remnant after distal gastrectomy. AB - BACKGROUND/AIMS: Helicobacter pylori infection is known to induce gastritis, oxidative stress, and cyclooxygenase (COX)-2 expression in the gastric mucosa. However, the effect of H. pylori infection on remnant gastritis has not been studied. We investigated whether the severity of remnant gastritis and COX-2 expression were affected by H. pylori infection after distal gastrectomy. METHODOLOGY: The study included 97 patients with gastric cancer who underwent curative distal gastrectomy with lymphadenectomy in our department between May 1999 and April 2001. All patients underwent endoscopic examination 2 weeks before and 12 weeks after surgery. The presence of H. pylori infection was determined by urease activity, hematoxylin-eosin staining, and immunochemical staining. Histologic remnant gastritis was graded based on the degree of neutrophil infiltration using the updated Sydney System. COX-2 expression was estimated immunohistochemically. RESULTS: Both the degree of neutrophil infiltration and the level of COX-2 expression were significantly higher in patients with than without H. pylori (p<0.05). There was a significant correlation between the degree of neutrophil infiltration and the degree of COX-2 expression (p<0.001). CONCLUSIONS: H. pylori eradication may become a treatment for preventing both remnant gastritis as well as remnant gastric carcinoma after distal gastrectomy. PMID- 15362791 TI - The expression of GAGE gene can predict aggressive biologic behavior of intestinal type of stomach cancer. AB - BACKGROUND/AIMS: There is still little information on the function of GAGE gene in stomach cancer except for cancer-specific gene expression recognized by autologous T lymphocytes. This study attempted to detect GAGE expression in stomach cancer to evaluate its clinical implication as a tumor marker or prognosticator as well as to find the candidate for immunotherapy. METHODOLOGY: Tumor samples from 60 patients and gastric juices from 18 patients with gastric cancer were studied by using RT-PCR with common primer. In-situ RT-PCR and southern blotting were performed to confirm the RT-PCR products. RESULTS: No expression of GAGE was observed in non-neoplastic juices and tissues. Fifteen out of 60 tumor tissues expressed GAGE (25.0%) mRNA, of which 13 cases (86.7%) were intestinal type and only 2 cases (13.3%) were diffuse type. In gastric juice, 22.2% (4/18) showed mRNA expression, all of which were intestinal type. GAGE expressions in both cancer tissues and gastric juices had a significant tendency to be higher by stage and lymph node metastasis (p<0.05). However, they did not show a significant relationship with tumor cell differentiation and vascular and perineural invasions. CONCLUSIONS: These results suggest that GAGE gene might have an important role in the development and progression of intestinal type of stomach cancer and the detection of GAGE mRNA may be eligible to the panel of molecular markers for aggressive behavior as well as being useful to a molecule for cancer-specific immunotherapy. PMID- 15362792 TI - Is bursectomy necessary for patients with gastric cancer invading the serosa? AB - BACKGROUND/AIMS: To confirm the impact of bursectomy on survival, we reviewed the clinical records of patients who underwent radical total gastrectomy with bursectomy for gastric cancer invading the serosa, with special reference to the location of tumor invasion. METHODOLOGY: From the records, patients were selected for this retrospective cohort study according to the following criteria: (a) Invasion beyond the serosal surface, (b) No metastases to liver, peritoneum, or distant organs, (c) Negative for peritoneal lavage cytology, and (d) Patients underwent curative D2 total gastrectomy with complete omental bursectomy. A total of 134 patients were eligible. These patients were divided into a group I which included patients with tumors that invaded only the posterior wall and a group II which included those with others. Survival was examined by uni- and multivariate analyses. RESULTS: Survival rates at 3 and 5 years were 67.3% and 53.0% for group I and 68.8% and 53.8% for group II. There was no significant difference in the survival between the two groups (p=0.969), even if survival was stratified by various clinicopathological factors. Multivariate analyses demonstrated that the significant independent factors for the survival were macroscopic type and lymph node metastasis. Location of the invasion was not a significant factor. CONCLUSIONS: These results suggested that bursectomy did not inhibit the spreading of tumor cells into the retro-stomach space. There might be no survival benefit of bursectomy in patients with gastric cancer. PMID- 15362793 TI - Survival of gastric cancer with concomitant liver metastases. AB - BACKGROUND/AIMS: Prognosis of gastric cancer with concomitant liver metastasis is poor. Gastrectomy and chemotherapy had been reported to be beneficial to this group of patients. Whether all the patients can benefit from that treatment modality and whether the clinical characteristics can give some information about survival have not been evaluated. METHODOLOGY: Eighty-three gastric cancer patients with concomitant liver metastases who had received treatment in the past 10 years were retrospectively studied with special reference to different extent of liver metastases. Clinical characteristics (peritoneal tumor dissemination, tumor markers, clinicopathological factors), treatment modalities (gastrectomy or chemotherapy) and survival were analyzed for their possible relationship with the extent of liver metastases. RESULTS: There were 33 patients with liver metastases limited to one lobe and 50 patients had metastases in both lobes. The clinicopathological characteristics show no difference between uni- and bilobar liver metastases except higher percentage of concomitant peritoneal dissemination in patients with bilobar disease. More unilobar involvement patients underwent gastrectomy (70% vs. 48%, p=0.018). Higher frequency of abnormal CEA level in bilobar metastases (38% vs. 60%, p=0.045). Median survival of unilobar metastasis is 7.8 months and 4.3 months for the bilobar involvement (p=0.001). Gastrectomy might prolong the survival in patients with unilobar metastasis (p=0.005), but not in patients with bilobar diseases (p=0.074). Chemotherapy could prolong the survival in patients without gastrectomy, but not the survival after gastrectomy. CONCLUSIONS: The clinicopathological characteristics and tumor markers have no role in predicting the survival among patients with liver metastases. Both gastrectomy and chemotherapy for the patients without gastrectomy could prolong survival, but chemotherapy could not prolong the survival after gastrectomy, the benefit of resection and chemotherapy are probably caused by selection bias. PMID- 15362794 TI - Evaluation of the effects of Helicobacter pylori eradication therapy on gastric antral epithelial hyperproliferation: a prospective six-month follow-up study. AB - BACKGROUND/AIMS: H. pylori-induced hyperproliferation of the gastric epithelium may have a critical role in gastric carcinogenesis. H. pylori-related hyperproliferation and reversibility of hyperproliferation after eradication therapy is still controversial. Therefore, we have evaluated the effects of H. pylori and its eradication on gastric antral epithelial proliferation. METHODOLOGY: A total of 32 H. pylori-positive and 22 H. pylori-negative subjects were enrolled into the study. Triple eradication therapy was given to the H. pylori-positive group. Upper endoscopy was repeated one month after the therapy and six months later, antral biopsy specimens were taken in each endoscopy. Biopsy specimens from H. pylori-negative subject were taken at the beginning of the study and sixth months later also. RESULTS: Proliferative index was 40.2% in H. pylori-positive state; it regressed to 27.6% after eradication and six months later the proliferative index was 30.7%. H. pylori-negative group's proliferative index was 25.5% initially and six months later it was 25.6%. The difference between the H. pylori-positive and -negative group was statistically significant (p<0.0001). The difference between H. pylori-positive group's values at the beginning of the study and one month after the eradication was significant (p<0.0001). In addition, the difference between H. pylori-positive group's initial values and those six months after eradication was also significant (p<0.0001). CONCLUSIONS: H. pylori increased the gastric epithelial proliferation and after the eradication therapy proliferative index decreased to control values. H. pylori and the related factors inducing gastric antral hyperproliferation may have an important role in H. pylori-related gastric malignancies. PMID- 15362795 TI - Laparoscopic and open approach in perforated peptic ulcer. AB - BACKGROUND/AIMS: To improve the perioperative course and quality of life of the patients with perforated peptic ulcer, we investigated the conversion risk factors in laparoscopic simple closure and omental patch repair for perforated peptic ulcer. METHODOLOGY: A total of 71 patients with a mean age of 46 years (rage, 17-81 years) was examined, and their clinical data was recorded and analyzed. RESULTS: The postoperative durations of nasogastric tube insertion, ileus, analgesic requirement, resuming diet, and hospital stay were shorter in the patients with laparoscopic simple closure than in those with open simple closure and omental patch repair. Univariate regression analysis revealed that the age, American Society of Anesthesiologist classification, presence of concomitant disease, and length of free air or fluid collection shown in abdominal computerized tomography significantly correlated with the conversion of laparoscopic simple closure and omental patch repair to open simple closure and omental patch repair. CONCLUSIONS: These conversion factors are useful to achieve appropriate surgical treatments for perforated peptic ulcer. PMID- 15362796 TI - The effect of H2-receptor antagonist and proton pump inhibitor on microbial proliferation in the stomach. AB - BACKGROUND/AIMS: Intra-gastric bacterial proliferation is frequent in patients with hypochlohydria. However, status of gastric bacterial infection in patients receiving proton pump inhibitor or H2-receptor antagonist remains controversial. The purpose of this study was to investigate the microbial condition of the stomach in patients who received H2-receptor antagonist or proton pump inhibitor. METHODOLOGY: Between November 2000 and January 2002, 102 patients were enrolled in this study. Of these, 52 did not receive any treatment (group I), 26 received H2-receptor antagonist (group II), and 24 received proton pump inhibitor (group III). Ten mL of gastric juice were aspirated for culture during endoscopic examination. The aerobic and anaerobic bacterial and fungal cultures were performed immediately. A glass pH meter measured the pH of the gastric juice. RESULTS: The intra-gastric pH was 2.91+/-2.06 (mean +/- SD), 4.12+/-2.83, and 5.11+/-2.47 for groups I, II, and III, respectively (p=0.001 between groups I and III, p>0.05 between groups I and II, and groups II and III). The positive bacterial culture rates were 66.7% (16/24) in group III, 46.2% (12/26) in group II, and 28.8% (15/52) in group I (p=0.007 between groups III and I,p>0.05 between groups I and II, and groups II and III). The positive candidal culture rates were 12.5% (3/24) in group III, 11.5% (3/26) in group II, and 17.3% (9/52) in group I (p>0.05). CONCLUSIONS: Patients who received proton pump inhibitor had more acid suppression and intra-gastric bacterial infection than those of the control group. The intra-gastric candidal infection was not related to intra-gastric pH or anti-secretory medication in this study. PMID- 15362797 TI - The use of tumor markers as predictors of prognosis in gastric cancer. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the prognostic significance of serum carcinoembryonic antigen (CEA) and carbohydrate antigen (CA19-9) levels in patients with gastric cancer. METHODOLOGY: During the period January 2000 and January 2003, 36 patients operated for primary gastric cancer, at Sisli Etfal Training and Research Hospital, Department of General Surgery, were analyzed. Serum CEA and CA19-9 levels were determined preoperatively and the correlation between the elevated levels of tumor markers and several clinicopathological features, and survival were evaluated. RESULTS: Elevated serum CEA and CA19-9 levels were determined in 10 of 35 patients (28.6%), and 9 of 31 patients (29%), respectively, and both markers were elevated in 3 of 31 patients (9.6%). Elevated levels of CEA correlated with depth of invasion (p=0.018) and pathological stage (p=0.029); elevated levels of CA19-9 correlated with lymph node metastasis (p=0.026); and elevated levels of both markers correlated well with lymph node metastasis (p=0.031). The survival of patients with normal CEA levels was significantly better than those with elevated levels (p=0.0072). CONCLUSIONS: Preoperative serum CEA and CA19-9 levels may add useful information in patients with gastric carcinoma, and CEA level is a predictor of prognosis. PMID- 15362798 TI - Duodenogastric reflux eradicates Helicobacter pylori after distal gastrectomy. AB - BACKGROUND/AIMS: Patients who undergo distal gastrectomy often develop duodenogastric reflux and preoperative H. pylori infection is eradicated spontaneously after distal gastrectomy in some patients. However, whether a causal relationship exists has not yet been studied. This report examines the correlation between H. pylori eradication and the amount of duodenogastric reflux following distal gastrectomy. METHODOLOGY: Among 72 consecutive patients who underwent curative distal gastrectomy with radical lymphadenectomy for gastric cancer, 37 patients had H. pylori infection preoperatively and were included in this study. The period of bile reflux (percent time) into the gastric remnant was measured with the Bilitec 2000 under standardized conditions on the 14th day after the surgery. Endoscopic examination was performed to determine the presence of H. pylori infection on week 12 after surgery. RESULTS: The percent time was higher in patients whose H. pylori infection had been eradicated after distal gastrectomy (58.1+/-9.2%) than in patients who had H. pylori infection after distal gastrectomy (33.8+/-5.7%). CONCLUSIONS: Duodenogastric reflux correlates with spontaneous eradication of H. pylori infection following distal gastrectomy. PMID- 15362799 TI - Helicobacter pylori infection in patients with ulcer recurrence after partial gastrectomy. AB - BACKGROUND/AIMS: Recurrent ulceration of the stomach occurs in some patients after surgery for peptic ulcer disease. The aim of this study is to evaluate the association of Helicobacter pylori infection with ulcer recurrence in patients after partial gastrectomy due to peptic ulcer disease. METHODOLOGY: A total of 186 patients after partial gastrectomy with Billroth I or Billroth II anastomosis presenting with dyspepsia or bleeding were included. An ulcer recurrence was documented by endoscopic examination. Biopsy specimens were taken from the remnant gastric bodies in all patients. H. pylori infection was diagnosed by either a positive biopsy urease test or the presence of the microorganism on histology. RESULTS: Eighty-three (44.6%) patients among the 186 patients had recurrent ulcers. H. pylori infection was found in 29 (36%) patients in the ulcer group and 42 (41%) patients in the non-ulcer group. The incidence of H. pylori infection did not differ significantly between the two groups (P>0.05). CONCLUSIONS: H. pylori infection may not play an important role in the pathogenesis of recurrent ulcer after partial gastrectomy. Other factors should be studied further. PMID- 15362800 TI - Effects of TS-1 on peritoneal dissemination of gastric cancer in nude mice. AB - BACKGROUND/AIMS: We investigated the effects of TS-1 on the survival of nude mice developing peritoneal dissemination of gastric cancer. METHODOLOGY: MKN-45 cells were injected into the peritoneal cavity of nude mice and a model of peritoneal dissemination was developed. TS-1 was administered orally every day from day 1 to day 10 or day 10 to day 19. RESULTS: Survival time of these treatment groups was significantly longer than untreated controls. In a pharmacokinetic study, TS-1 was administered on day 10 and the 5-fluorouracil levels were retained and maintained for a longer time, in the ascites and tumor than in plasma. The area under the concentration curve for 5-FU in the tumor was higher, than in plasma or ascites. CONCLUSIONS: TS-1 could be effective in treating peritoneal dissemination of gastric cancer, due to the supply of 5-fluorouracil in the tumor by systemic and intraperitoneal circulation. PMID- 15362801 TI - Geranylgeranylacetone attenuates suppression by Helicobacter pylori extract of human umbilical vein epithelial cell growth. AB - BACKGROUND/AIMS: Helicobacter pylori infection delays gastric ulcer healing. Angiogenesis is important for the healing of gastric ulcers. Therefore, the effects of H. pylori water extract and a novel antiulcer drug, geranylgeranylacetone, on the viability of human umbilical vein epithelial cells (HUVECs) were investigated. METHODOLOGY: H. pylori (ATCC43504) was prepared by sonication. The HUVEC viability after treatment with H. pylori water extract alone or in combination with geranylgeranylacetone was estimated by an MTT assay. RESULTS: H. pylori water extract significantly decreased cell viability in a concentration-dependent manner after 48 h. However, combined use of H. pylori water extract and geranylgeranylacetone significantly increased the HUVEC viability over that of H. pylori extract alone. CONCLUSIONS: A novel antiulcer drug, geranylgeranylacetone, attenuates the H. pylori-induced inhibition of angiogenesis. PMID- 15362802 TI - Postoperative evaluation of the jejunal pouch reconstruction following proximal and distal gastrectomy for cancer. AB - BACKGROUND/AIMS: To evaluate the efficacy of the jejunal pouch reconstruction following subtotal proximal and distal gastrectomy, a retrospective study examining the postoperative condition of patients who underwent different methods of reconstruction after gastrectomy for cancer was undertaken. METHODOLOGY: Various parameters indicative of postoperative function were evaluated at one year postoperatively, and two major groups were examined. The "proximal gastrectomy" group was composed of patients who underwent either 1) proximal gastrectomy with an interposed jejunal pouch (PG-pouch), 2) proximal gastrectomy with simple jejunal interposition (PG-inter), or 3) total gastrectomy with simple jejunal interposition (TG). The "distal gastrectomy" group was composed of patients who underwent either 4) distal gastrectomy with an interposed jejunal pouch (DG-pouch), 5) distal gastrectomy with simple jejunal interposition (DG inter), or 6) distal gastrectomy with Billroth 1 reconstruction (B-1). RESULTS: Volume of meal intake was better preserved and the incidence of abdominal symptoms were less frequent in the PG-pouch and DG-pouch groups. In the PG-inter, DG-inter and DG-pouch groups, none of the patients experienced heartburn or had endoscopic findings consistent with reflux esophagitis, while 2 patients (20.0%) in the PG-pouch group complained of heartburn with evidence of reflux esophagitis on endoscopy. Increase in blood acetaminophen level was milder in both the PG pouch and DG-pouch groups, signifying improved gastric emptying. CONCLUSIONS: The jejunal pouch interposition following proximal and distal gastrectomy seems to confer clinical benefit in terms of postoperative function, especially in the form of meal intake, abdominal symptoms, and gastric emptying. The side effect of an improved reservoir may be the incidence of reflux esophagitis seen in 2 patients in the PG-pouch group. PMID- 15362803 TI - Long-term survivor of gastric small cell carcinoma. AB - We describe the long-term survival of a patient following the diagnosis of primary gastric small cell carcinoma. In January 2000, a 73-year-old male was found to have advanced gastric small cell carcinoma directly invading his liver. He received combination chemotherapy with cisplatin and irinotecan as first-line chemotherapy, then cisplatin and etoposide as second-line chemotherapy. He had a complete response after four cycles of second-line chemotherapy. In March 2001, the tumor recurred in the stomach and the patient underwent a total gastrectomy. He has survived free of disease for more than 2 years after the first diagnosis. PMID- 15362804 TI - A case of isolated pancreatic metastasis of gastric cancer presenting problematic discrimination from gastropancreatic double cancer. AB - We experienced a case of isolated pancreatic metastasis caused by gastric cancer which showed high levels of the tumor markers relevant to pancreatic cancer. The patient was a 59-year-old man who had tumors in both the gastric antrum and pancreatic head. He was diagnosed as having double cancer of the stomach and pancreas on account of the high values of pancreatic cancer-associated markers, and underwent operation. This resulted in non-curable resection attributable to broad lymph node metastasis. The resected specimen lacked continuity between the pancreas tumor and gastric tumor, and the pancreatic tumor was histologically diagnosed as metastasis from the gastric cancer. Moreover, the advanced lymph vessel invasion suggested possible metastasis through the lymph ducts. The present case is a rare metastatic form because there has been no report of lymphogenous isolated pancreatic metastasis of gastric cancer. In advanced gastric cancer, the possibility of pancreatic metastasis should be always borne in mind, and pancreatic biopsy may be taken into account at times. Radical operation may be performed corresponding to the degree of each stage of the double cancer of stomach and pancreas. However, considering the low possibility of curable resection for pancreatic metastasis of gastric cancer, other treatments than surgery should also be considered. PMID- 15362805 TI - Rational basis for optimizing antibiotic dosing regimens. AB - Adjustment of the antibiotic dosage is usually done taking into account pharmacokinetic parameters. However, as the bacterial response to the antimicrobial effect varies it is important to correlate pharmacokinetics with antimicrobial susceptibility data, the latter measured by determining the Minimum Inhibitory Concentration (MIC). It is now widely accepted that the ratio between the maximum antibiotic concentration achieved in serum and the MIC of the pathogen correlates with efficacy of aminoglycosides and fluoroquinolones. The ratio between the area under the serum concentration-time curve and the MIC correlates with efficacy of aminoglycosides and fluoroquinolones but also of vancomycin, tetracyclines, azithromycin, and quinupristin/dalfopristin. Finally the time for which antibiotic concentration in serum remains above the MIC correlates with efficacy of beta-lactams, most macrolides, and clindamycin. All the above mentioned pharmacodynamic parameters should be considered for optimizing antibiotic dosage. PMID- 15362806 TI - Cell cycle--molecular mechanisms and therapeutic perspectives in oncology. AB - Regulation of cell division is an object of intensive studies for the needs of tumor biology and medicine. Cyclins and cyclin-dependent kinases are the major regulatory complexes that control the transition from one phase of the cell cycle to the following one. It has been established that the activity of certain cyclins is impaired in some tumors due to mutation of the coding gene. Verification of the specific change in a particular cyclin is discussed as a reliable diagnostic marker in clinical oncology. Mutations in genes for inhibitors of cyclin-dependent kinases correspond with definite tumor types. Impairment of control mechanisms leads to independence of cell division from signal regulators, which lies at the basis of tumorigenesis. Malignant transformation could result from overexpression of positive regulators like cyclins or loss of negative regulators like the inhibitors of the cyclin dependent kinases. These molecules are an object of the new chemo- and gene therapeutical antitumor strategies. The present review focuses on the main regulatory points in cell cycle phases and on the pharmacological agents that affect them. Novel antitumor strategies based on the influence on the molecular mechanisms regulating the cell cycle are discussed. PMID- 15362807 TI - Effect of mycophenolate mofetil (Cell Cept) in the treatment of immune glomerulopathies. AB - AIM: To study the effect of mycophenolate mofetil (Cell Cept) in treating patients with various types of chronic glomerulonephritis and other immune nephropathies. MATERIAL AND METHODS: Between 2000 and 2003 we treated 35 patients (18 women, 17 men) with Cell Cept (La Roche). In 32 patients the diagnosis was confirmed by kidney biopsy (immunofluorescence, light and electron microscopy). RESULTS: Treatment with Cell Cept was very successful in 22 of the patients in the study (62.86%). Proteinuria was significantly reduced and firmly maintained well below 0.5 g/l; serum protein levels were elevated to normal values, the edemas disappeared. In 12 patients the drug had a good effect: there was a significant reduction of proteins in the urea within 1.2 - 2.0 g/l, an increase of total protein and albumins in plasma but after three months of treatment. The therapy was with no effect only in one patient with primary amyloidosis of kidneys. CONCLUSIONS: Treatment with mycophenolate mofetil (Cell Cept) is an alternative modality for the management of immune glomerulopathies resistant to conventional and pulse pathogenetic therapeutic regimens. It can be a treatment of first choice. PMID- 15362808 TI - Placental soluble fms-like tyrosine-kinase-1 (sFlt-1) in pregnant women with preeclampsia. AB - The pathophysiology of preeclampsia remains largely unknown. A number of circulating placenta-produced factors have been implicated in causing the endothelial dysfunction and the clinical phenotype characteristic of preeclampsia. AIM: Determination of serum levels of placental soluble fms-like tyrosine-kinase-1 (sFlt-1) in pregnant women with preeclampsia. Eleven pregnant women with preeclampsia and 11 healthy women (controls) were included in the study. Determination of sFlt-1 was done with ELISA. The mean serum sFlt-1 levels of pregnant women with preeclampsia were twice as high as that of women with normal pregnancy. The highest level of sFlt-1 was found in women with severe preeclampsia. In women with mild form of preeclampsia the sFlt-1 level was close to that of the controls. sFlt-1 appears to be involved in the pathogenesis of preeclampsia and its serum levels can be used as a diagnostic marker of preeclampsia. PMID- 15362809 TI - Nasal reconstruction with vascularized forehead flap (preliminary communication). AB - AIM: The authors have studied the anatomical characteristics of the vascularization of the forehead flap used for nasal reconstruction. MATERIAL AND METHODS: For the period 1990--2000 ten cadaver dissections were performed and three patients underwent reconstructions using the forehead flap. The results obtained reveal that the blood supply of the flap is provided by the suprathrochlear, supraorbital and frontal branch of the superficial temporal vessels, which form a network of anastomoses between the frontal muscle and the skin. Three cases of successful nasal reconstruction are presented. Two of the patients had traumatic injury of the nose and one was with postoperative defect in the naso-labial area, nose and maxilla obtained after ablation surgery for neoplasm (spinocellular carcinoma). The reconstruction was done with vascularized oblique forehead flap. DISCUSSION: The results were evaluated as good. There were no complications. CONCLUSION: The results of the anatomical study of the blood supply of the forehead flap and the clinical results of nasal reconstruction are discussed in relation with the literature data. PMID- 15362810 TI - Operative treatment of postpulmonectomy empyema. AB - OBJECTIVE: To analyse retrospectively 10 patients with postpulmonectomy empyema. METHODS: Ten patients (9 men, 1 woman, aged 48 to 69 years) with postpulmonectomy empyema treated over a 12 year period (1991--2002) were analysed. Nine of the patients had pulmonectomy for lung carcinoma and one--for pulmonary abscess. All postpulmonectomy empyemas were managed in two stages--an open-window thoracostomy for curing the infective process in the empyema cavity and then closure of the bronchopleural fistulas and obliteration of the cavity. Video-assisted thoracoscopy (VATS) was successfully used by the authors for sterilization and debridement of the cavity, and finally to insert two thoracic drains for the cavity lavage. The obliteration of the empyema cavity was achieved by a vascular muscle flap transposition from the thoracic muscles. The latissimus dorsi muscle flap was used in 6 patients. The pectoralis major muscle flap was used in 4 patients. Additionally, a thoracoplasty was performed in the patients with pectoralis major muscle flap transposition. RESULTS: Eight patients (80%) were discharged from hospital without complications, and two patients died (20%). CONCLUSIONS: On the basis of the study results and the literature review the authors conclude that the applied therapy yields good results in the treatment of postpulmonectomy empyema. PMID- 15362811 TI - Optimal computed tomographic (CT) examination technique for microendoscopic sinus surgery. AB - AIM: Functional endoscopic sinus surgery is widely utilized to assess and treat inflammatory sinonasal diseases. The use of this surgical technique has resulted in a dramatic increase of the radiologic studies carried out as part of presurgical preparation. Computed tomography (CT) has become a standard for pre endoscopic and postoperative assessment of the paranasal sinuses. In our study we aimed at finding the optimal imaging technique for CT examination of the sinuses. MATERIALS AND METHODS: We examined 60 patients with inflammatory sinonasal diseases using Sytec 3000 CT scanner. The scans were performed in the coronal and axial plane. The kilovolt peak was kept constant and the milliampere second setting was reduced without image compromise. CONCLUSION: CT study is a method of choice in diagnosing inflammatory diseases of the sinuses. The technique we suggest in the study provides maximal diagnostic information at reduced cost and radiation exposure of the patient. PMID- 15362812 TI - Vertical ramus osteotomy for treatment of mandibular prognathism. AB - Following a brief literature review of treatment modalities for mandibular prognathism the authors propose a simplified method for vertical ramus osteotomy. The method was successfully used in 24 patients with mandibular prognathism (alone or in combination with open bite). Postoperative complications (suppuration of the operative wound) were found in 2 patients (8%), and postoperative recurrences--in 5 patients (20.8%). No complications, usually associated with sagittal and vertical intraoral osteotomies (hypoesthesia of the trigeminal nerve, Frey's syndrome, dissecting aneurysm of the internal carotid artery) were found to be consequential to the proposed method. PMID- 15362813 TI - Effects of kyotorphin and analogues on nociception and pentylenetetrazole seizures. AB - Kyotorphin (Kyo) is synthesized in specific brain regions where it may modulate synaptic transmission and directly excites cortical neurons, and indirectly exerts opioid actions to produce analgesia via release of met-enkephalin. Kyo is formed by specific enzyme from L-tyrosine and L-arginine in the presence of ATP and Mg2+ in the brain. Kyo and its analogues Tyr-Cav, Tyr (Cl2)-Cav exerted naloxone-reversible antinociception by paw-pressure test. Kyo exerted anticonvulsive effect on the pentylenetetrazole (PTZ) seizure model. AIM: To investigate the analgesic and the anticonvulsive effects of Kyo, Tyr-Cav and Tyr(Cl2)-Cav during acute pain and PTZ seizure model. METHODS: Changes in the nociceptive effects were examined in male Wistar rats by the tail flick (TF) and hot plate (HP) tests. Kyo, Tyr-Cav, Tyr(Cl2)-Cav were applied in rats intracerebroventricularly (i.c.v.) at a dose of 20 microg/20 microl. The anticonvulsive effects of peptides were studied on a PTZ seizure model. The peptides were applied in male mice at a dose of 20 microg/mouse (i.c.v.). RESULTS: Kyo, Tyr-Cav, Tyr(Cl2)-Cav exerted analgesic effects in both nociceptive tests used. The effects were more pronounced for L-Arg, L-Cav, Tyr-Cav and Tyr(Cl2)-Cav. In PTZ seizure model Kyo and its analogues exerted strong inhibition on seizure intensity compared with control group. CONCLUSION: Taken together, these results reveal Kyo, Tyr-Cav and Tyr(Cl2)-Cav as a behaviorally active peptide in experimental animal models. PMID- 15362814 TI - A transitory polycytaemia after parathyroidectomy for secondary hyperparathyroidism--a literature survey and a case study. AB - The present study is a literature review of a nephrological problem--secondary hyperparathyroidism, its surgical treatment and the effect of the intervention on the anemic syndrome in patients with chronic renal failure (CRF) on hemodialysis treatment. We present a case of a 29-year-old female hemodialysis patient with a transitory polycytaemia (duration of 5 months) after parathyroidectomy for secondary hyperparathyroidism. Prior to surgery she was constantly anemic in spite of epoetin treatment lasting 10 years. Ten days after the parathyroidectomy there was a crucial decline of parathormone (from 900.4 pg/ ml to 21.4 pg/ml) and ionized calcium (iCa) (from 1.23 mmol/l to 0.79 mmol/l) and the hemoglobin level reached 170 g/l with a hematocrit of 0.53. After discontinuation of epoetin treatment (58.8 U/kg/week) in the following 5 months hemoglobin level declined reaching 144 g/l. Two years after parathyroidectomy the patient is still with normal hemoglobin level (125-135 g/l) without epoetin treatment. We discuss the likely mechanisms for the early onset of polycytaemia after parathyroidectomy as well as the importance of adequate control of secondary hyperparathyroidism in the management of anemia in chronic renal insufficiency. PMID- 15362815 TI - Secondary poisoning of eagles following intentional poisoning of coyotes with anticholinesterase pesticides in western Canada. AB - Records of eagles, coyotes (Canis latrans), and red foxes (Vulpes vulpes) necropsied at the Western College of Veterinary Medicine, Saskatoon, Saskatchewan, Canada, between 1967 and 2002 were reviewed for cases suggestive of anticholinesterase poisoning. From 1993 to 2002, 54 putative poisoning incidents involving 70 bald eagles (Haliaeetus leucocephalus) and 10 golden eagles (Aquila chrysaetus) were identified. Of these, 50 incidents occurred in Saskatchewan, two were in Manitoba, and one occurred in each of Alberta and the Northwest Territories. The diagnosis was confirmed in eight instances by demonstration of pesticide in ingesta from eagles or known use of pesticide at the site together with brain cholinesterase (AChE) reduction of >50% in at least one animal. A presnmptive diagnosis of poisoning was made in 33 incidents based on brain AChE reduction of >50% in at least one animal; 13 incidents were considered suspicious because of circumstantial evidence of the death of eagles in association with other species and limited AChE reduction. Other wild species were found dead in 85% of the incidents involving eagles. Coyotes, foxes, black-billed magpies (Pica pica), and striped skunks (Mephitis mephitis) were associated with 34, six, six, and three incidents, respectively. There were eight additional incidents that did not involve eagles in which poisoning was diagnosed in coyotes. Carbofuran was identified in nine incidents. Carbamate poisoning was indicated on the basis of reactivation of brain AChE activity in two additional incidents. Brain AChE activity was not reduced from normal in eagles in four of seven incidents in which carbofuran was identified. The organophosplorous insecticide terbufos was found together with carbofuran in one incident. Brain AChE activity was measured in wild canids and in eagles in 15 incidents; in all of these incidents, brain AChE was redulced by >50% in at least one mammal, whereas this level of reduction occrred in eagles in only four incidents. Use of anticholinesterase pesticides to poison coyotes is illegal, but the practice continues and secondary poisoning of eagles is a problem of unknown proportions in western North America. PMID- 15362816 TI - Effects of heavy metals on immunocompetence of white-footed mice (Peromyscus leucopus). AB - Continued human population growth and industrialization result in increased contamination of wildlife habitats. Effects of such habitat deterioration on the well-being of natural populations are unclear. Exposure to contaminants may impair immunocompetence, thereby increasing disease susceptibility. The mammalian immune system is important in maintaining health and in its sensitivity to toxins. In our study conducted from May 1999 through May 2001, we examined assays of immnnoompetence in the white-footed mouse (Peromyscus leucopus) that inhabited reference sites and sites significantly contaminated with mixtures of heavy metals. We estimated potential exposure and uptake of heavy metals by measuring the level of each contaminant in representative soil and tissue samples. Intraindividual variation across mice, but not sex, explained a large portion of the overall variance in immune response, and spleen weight was significanltly afflicted by mouse age. We found no evidence that residence on contaminated sites had any effect on immunopathology and humoral immunity as measured in our study. We suggest that field and laboratory studies in ecotoxicology provide estimates of exposure to contaminants (i.e., tissue analyses) to establish a database suitable to clarify the dose-response relationship between contaminants and target systems. PMID- 15362817 TI - Oral disease in free-living red squirrels (Sciurus vulgaris) in the United Kingdom. AB - Ninety-one red squirrels (Sciurus vulgaris) found dead in the UK between January 1994 and August 1998 were necropsied at the Institute of Zoology (London, UK); their oral cavities were examined visually, and in eight cases, radiographically. Four red squirrels, which had evidence of oral disease when necropsied as part of a mortality survey, also were examined. A low prevalence (prevalence = 0.033, SE = 0.02, n = 91) of oral disease was found in free-living red squirrels. In only two cases was oral disease the probable cause of death. Attrition of the check teeth (three cases) and overgrowth of the incisors (four cases) were the most common lesions found. Partial anodontia was recorded in one squirrel. PMID- 15362818 TI - Muskox lungworm (Umingmakstrongylus pallikuukensis) does not establish in experimentally exposed thinhorn sheep (Ovis dalli). AB - Muskoxen (Ovibos moschlatus moschatus) on the northwestern mainland of Nunavut and Northwest Territories, Canada, are infected with the protostrongylid lungworm, Umingmaksrongylus pallikuuhkensis. The geographic range of this muskox population is expanding to the south and west, and it is anticipated that these animals will eventually become sympatric with Dall's sheep (Ovis dalli dalli) in the Mackenzie and Richardson Mountains. To address the concern of wildlife managers that U. pallikulkensis may infect and adversely affect Dalls sheep, four Dalls/Stone's (Ovis dalli stonei) hybrid lambs and one adult muskox (Ovibos moschatus wardi) were each given 100 third-stage larvae of U. pallikuukensis. All animals were intensively monitored for 9 mo postinfection (PI) using clinical examinations, fecal analyses, hematology, blood chiemistry and medical imaging. No first-stage larvae of U. pallikuinkensis were recovered from the lambs and monitoring revealed nio evidence that the parasite had established in any of these animals. First-stage larvae were found in the feces of the muskox beginning at 94 days PI, and typical parasite cysts were visible in lung radiographs at 188 days PI. This study addresses an important management and wildlife health issue associated with the potential for host-switching of pathogens and indicates that it is improbable that thinhorn sheep are suitable hosts for U. pallikuukensis. PMID- 15362819 TI - Sylvatic plague reduces genetic variability in black-tailed prairie dogs. AB - Small, isolated populations are vulnerable to loss of genetic diversity through in-breeding and genetic drift. Sylvatic plague due to infection by the bacterium Yersinia pestis caused an epizootic in the early 1990s resullting in declines and extirpations of many black-tailed prairie dog (Cynomys ludovicianus) colonies in north-central Montana, USA. Plague-induced population bottlenecks may contribute to significant reductions in genetic variability. In contrast, gene flow maintains genetic variability within colonies. We investigated the impacts of the plague epizootic and distance to nearest colony on levels of genetic variability in six prairie dog colonies sampled between June 1999 and July 2001 using 24 variable randomly amplified polymorphic DNA (RAPD) markers. Number of effective alleles per locus (n(e)) and gene diversity (h) were significantly decreased in the three colonies affected by plague that were recovering from the resulting bottlenecks compared with the three colonies that did not experience plague. Genetic variability was not significantly affected by geographic distance between colonies. The majority of variance in gene fieqnencies was found within prairie clog colonies. Conservation of genetic variability in black-tailed prairie dogs will require the preservation of both large and small colony complexes and the gene flow amonog them. PMID- 15362820 TI - Ecogeographic patterns of rabies in southern Ontario based on time series analysis. AB - We describe a method based on time series analysis that divided the rabies enzootic area of southern Ontario into 13 regions using data collected at the township level, the smallest available geographical unit for Ontario (Canada). The intent was to discover ecogeographic patterns if such existed. For the period 1957-89, the quarterly time series of fox rabies cases for each of the 423 townships in the study area was correlated with the time series of its adjacent neighbors. Townships were then linked to adjacent townships provided the pair wise correlations had significant correlation coefficients. This procedure produced 13 clusters that remained stable when additional lead/lag relationships between townships were examined. Furthermore, those clusters, which we then termed "rabies units," had different behaviors in terms of species distribution, persistence, and periodicity. Time series in adjacent units were not synchronous. We discuss how our findings influenced the rabies control program in Ontario, how they relate to recent findings about the distribution of fox rabies virus subtypes, and how they lend support for the role of metapopulation structulre in persistence of disease. PMID- 15362821 TI - Exposure time of oral rabies vaccine baits relative to baiting density and raccoon population density. AB - Oral rabies vaccination (ORV) baiting programs for control of raccoon (Procyon lotor) rabies in the USA have been conducted or are in progress in eight states east of the Mississippi River. However, data specific to the relationship between raccoon population density and the minimum density of baits necessary to significantly elevate rabies immunity are few. We used the 22-km2 US National Aeronautics and Space Administration Plum Brook Station (PBS) in Erie County, Ohio, USA, to evaluate the period of exposure for placebo vaccine baits placed at a density of 75 baits/km2 relative to raccoon population density. Our objectives were to 1) estimate raccoon population density within the fragmented forest, old field, and industrial landscape at PBS: and 2) quantify the time that placebo, Merial RABORAL V-RG vaccine baits were available to raccoons. From August through November 2002 we surveyed raccoon use of PBS along 19.3 km of paved-road transects by using a forward-looking infrared camera mounted inside a vehicle. We used Distance 3.5 software to calculate a probability of detection function by which we estimated raccoon population density from transect data. Estimated population density on PBS decreased from August (33.4 raccoons/km2) through November (13.6 raccoons/km2), yielding a monthly mean of 24.5 raccoons/km2. We also quantified exposure time for ORV baits placed by hand on five 1-km2 grids on PBS from September through October. An average 82.7% (SD = 4.6) of baits were removed within 1 wk of placement. Given raccoon population density, estimates of bait removal and sachet condition, and assuming 22.9% nontarget take, the baiting density of 75/ km2 yielded an average of 3.3 baits consumed per raccoon and the sachet perforated. PMID- 15362822 TI - Detection of TT virus among chimpanzees in the wild using a noninvasive technique. AB - Zoonotic transmission and emergence of pathogens are serious threats to endangered populations of free-ranging primate species. Recent discovery of a nonpathogenic yet highly prevalent virus in human populations, TT virus (TTV), has prompted studies into the presence of this virus among captive individuals of other species of nonhuman primates. In this study, we screened captive primate species for TTV. In addition, we provide the first data on TTV infectionin free ranging primates by noninvasive screening of three chimpanzee (Pan troglodytes sweinfurthii) commlunities. Phylogenetic relationships between virus isolates and those previously reported from hulman popullations, captive primates, and domesticated species are inferred. Our findings are discussed with respect to potential zoonotic events that may result from increased levels of human encroachlment into wild habitats. PMID- 15362823 TI - Corticosteroid treatment does not reactivate canine herpesvirus in red foxes. AB - To study canine herpesvirus (CHV) reactivation from red foxes (Vulpes vulpes), 29 foxes with varying CHV antibody and CHV carrier status were treated with methylprednisolone acetate, a glucocorticosteroid drug with prolonged immunosuppressive effect in dogs. In the first experiment, 17 foxes with unknown CHV carrier status were treated once with methylprednisolone: in the second experiment, five foxes were treated twice, 4 mo after being intravenously CHV infected; and in the third experiment, six foxes were treated five times, 11 mo after peroral CHV infection. Infectious CHV was not isolated after treatment from either naturally or experimentally CHV-infected foxes or from untreated, CHV seronegative in-contact foxes. Canine herpesvirus DNA was not detectable in mucosal secretions or white blood cells of any of the foxes, whereas all trigeminal ganglia of experimentally CHV-infected foxes were polymerase chain reaction-positive. In CHV-seropositive foxes, anti-CHV antibody titers did not change with time after treatment, and CHV-seronegative in-contact controls did not seroconvert. Hematologic parameters remained mostly unchanged. We conclude that CHV is not as easily reactivated in foxes following corticosteroid treatment as in dogs, although there was no obvious sign of immunosuppression. Canine herpesvirus was not spread from virus carriers to naive in-contact foxes, which may be among possible explanations for the reported low CHV prevalence in wild foxes. PMID- 15362824 TI - Use of recombinant antigens of Borrelia burgdorferi and Anaplasma phagocytophilum in enzyme-linked immunosorbent assays to detect antibodies in white-tailed deer. AB - Serum samples obtained from white-tailed deer (Odocoileus virginianus) in Connecticut (n=218) and South Carolina (n=20) (USA) during the period 1992-2002 were analyzed for antibodies to whole-cell or recombinant antigens (i.e., fusion proteins) of Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum, etiologic agents of Lyme borreliosis and granulocytic ehrlichiosis, respectively. In enzyme-linked immunosorbent assays (ELISAs) with whole-cell B. burgdorferi, the overall seropositivity rate for Connecticut (53%) exceeded that for South Carolina (30%). In separate tests of seven recombinant antigens of B. burgdorferi by an ELISA, seroprevalence for the VlsE antigen was highest (48%) in Connecticut followed by outer surface protein (OspF) (21%), whereas serum reactivities to the protein (p) 41-G antigen (55%) and VlsE (25%) were most frequent for South Carolina sera. In analyses for antibodies to the recombinant protein (p) 44 antigen of A. phagocytophilum, seroprevalences of 52% and 25% were recorded for Connecticut and South Carolina samples, respectively. These findings paralleled those determined by indirect fluorescent antibody staining methods with whole cells (43% and 30%). Moreover, there was good agreement (74%) in results of Western blot analyses and an ELISA when a subset of 39 sera was screened with whole-cell or recombinant p44 antigens of A. phagocytophilum. An ELISA with highly specific recombinant VlsE or p44 antigens can be used in conjunction with other antibody tests to determine whether deer living in different regions of eastern United States were exposed to B. burgdorferi or A. phagocytophilum. PMID- 15362825 TI - Evidence for chronic stress in captive but not free-ranging cheetahs (Acinonyx jubatus) based on adrenal morphology and function. AB - The cheetah (Acinonyx jubatus) is highly endangered because of loss of habitat in the wild and failure to thrive in captivity. Cheetahs in zoos reproduce poorly and have high prevalences of unusual diseases that cause morbidity and mortality. These diseases are rarely observed in free-ranging cheetahs but have been documented in cheetahs that have been captured and held in captive settings either temporarily or permanently. Because captivity may be stressful for this species and stress is suspected as contributing to poor health and reproduction, this study aimed to measure chronic stress by comparing baseline concentrations of fecal corticoid metabolites and adrenal gland morphology between captive and free-ranging cheetahs. Additionally, concentrations of estradiol and testosterone metabolites were quantified to determine whether concentrations of gonadal steroids correlated with corticoid concentration and to assure that corticosteroids in the free-ranging samples were not altered by environmental conditions. Concetntrations of fecal corticoids, estradiol, and testosterone were quantified by radioimmunoassay in 20 free-ranging and 20 captive cheetahs from samples collected between 1994 and 1999. Concentrations of baseline fecal corticoids were significantly higher (p = 0.005) in captive cheetahs (196.08 +/- 36.20 ng/g dry feces) than free-ranging cheetahs (71.40 +/- 14.35 ng/g dry feces). Testosterone concentrations were lower in captive male cheetahs (9.09 +/- 2.84 ng/g dry feces) than in free-ranging cheetahs (34.52 +/- 12.11 ng/g dry feces), which suggests suppression by elevated corticoids in the captive males. Evidence for similar sulppression of estradiol concentrations in females was not present. Adrenal corticomedullary ratios were determined on midsagittal sections of adrenal glands from 13 free-ranging and 13 captive cheetahs obtained between 1991 and 2002. The degree of vacuolation of cortical cells in the zona fasciculata was graded for each animal. Corticomedullary ratios were larger (p = 0.05) in captive cheetahs; however, there was no difference (p = 0.31) in the degree of corticocyte vacnolation between the two populations. These data proxile both mnorphologic and functional evidence suggestive of chronic stress in captive cheetahs. Further research into the role of hypercortisolemia in the pathogenesis of the reproductive abnormalities and unusual diseases of captive cheetahs is needed. PMID- 15362826 TI - Levels of fecal corticosterone in sandhill cranes during a human-led migration. AB - Fourteen captive-reared greater sandhill cranes (Grus canadensis tabida) were conditioned to follow ultralight aircraft to promote migration between Wisconsin and Florida (USA) after release. Fecal samples were collected throughout the training period in Wisconsin and during a l977-km human-led migration to Florida to determine fecal corticosterone (FC) concentrations by radioimmunnoassay. The mean (+/-SE) FC concentration during the training period was 109.5 +/- 7.5 ng/g and was representative of baseline levels recorded previously from sandhill cranes. Fecal corticosterone concentrations increased in early migration compared to concentrations I mo prior to departure (P < 0.01) but were not different from baseline concentrations at tile end of the 6-wk migration period. The variability of FC concentrations in individual samples was greater throughout the migration than the training period. Increases in FC during migration were modest and generally consistent with normal corticosterone elevations observed in migrating birds. PMID- 15362827 TI - Comparison of three fecal steroid metabolites for pregnancy detection used with single sampling in bighorn sheep (Ovis canadensis). AB - We conpared three fecal steroid metabolite assays for their usefulness in detecting pregnalcy among free-ranging Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from Bighorn Canyon National Recreation Area, Wyoming and Montana (USA) and captive bighorn ewes at ZooMontana in Billings, Montana. Fecal samples were collected from 11 free-ranging, radio-collared bighorn ewes in late January-May 2001 and from 20 free-ranging, radio-collared ewes in late March to mid-May 2002. Free-ranging ewes were monitored the following spring to determine whether or not they lambed. In addition, two captive ewes were studied at ZooMontana. With three exceptions, free-ranging bighorn ewes that produced lambs had nonspecific progesterone metabolite (iPdG) levels of >1800 ng/g feces and iPdG levels >7000 ng/gm feces when samples were collected between early March and mid-May. Samples collected earlier in the year were inconclusive. One false negative was suspected to be the result of sample collection error. Of the captive ewes, nonspecific pregnanediol-3alpha-glucuronide (PdG) and iPdG followed a predictable curve over the course of the 180-day pregnancies. We conclude that estrone conjugates are not useful in diagnosing pregnancy; however, fecal steroid analysis of PdG and iPdG can be used to accurately determine pregnancy and reproductive function in bighorn sheep. This holds great potential as a noninvasive technique for understanding the role of reproductive disease in wild bighom sheep. PMID- 15362828 TI - Immobilization of mule deer with thiafentanil (A-3080) or thiafentanil plus xylazine. AB - We evaluated thiafentanil oxalate (A-3080) for the immobilization of mule deer (Odocoileus hemionus) under laboratory and field conditions. In a crossover experiment comparing recommended (0.1 mg/kg) and 2x recommended thiafentanil doses in captive deer, both produced rapid induction and immobilization. Mean induction was shorter (P = 0.013) for the 2x group (1.9 vs. 3 min); mean reversals for both groups were rapid (recommended = 0.9 min after naltrexone injection; 2x = 1 min) and did not differ (P = 0.29). Six free-ranging mule deer were immobilized with 7 mg thiafentanil and four with 10 mg; mean induction was 2.3 min for both groups (95% confidence interval [CI]: 7 mg, 1.2-3.4; 10 mg, 1.9 2.8), and mean reversal was <1 min for both groups. Of 165 free-ranging deer darted with various combinations of thiafentanil and xylazine, we successfully immobilized 148 (90%). Mean induction ranged from 2.1 to 4.9 min for different drug combinations. Reversals were not compared because naltrexone and yohimbine doses varied, but overall mean reversal was 1.9 min (95% CI, 1.7-2.1 min) after injection of naltrexone and yohimbine intravenously (i.v.); naltrexone:thiafentanil ratios ranging from 10:1 to 43:1 provided mean recoveries ranging from 1.5 to 2.3 min. All 25 deer fitted with radio collars were alive at 30 days postcapture. On the basis of overall reliability and effectiveness, drug volumes, and ease of handling drugged animals, we recommend using a combination of 10-12 mg thiafentanil (0.15-0.2 mg/kg) and 100 mg xylazine to immobilize mule deer; immobilization can be effectively reversed with 100 mg naltrexone or more and 15 mg yohimbine or more i.v. Where feasible, we also recommend the use of transmitter darts when immobilizing mule deer with opioids in order to maximize recovery of darted deer and to ensure that missed darts are found. PMID- 15362829 TI - Hemorrhagic disease in Kansas: enzootic stability meets epizootic disease. AB - Kansas (USA) could represent a transition area between contrasting epidemiologic patterns of hemorrhagic disease (HD) in the midwestern United States. In this study, we compare the distribution of reported clinical HD with serologic data to determine whether the risk of HD in white-tailed deer (Odocoileus virginianus) is associated with geographic location corresponding to the reported distribution of two white-tailed deer subspecies. On the basis of a high prevalence of antibodies (91-100%) to multiple serotypes of epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV), with correspondingly few reports of clinical HD, it appears that a state of enzootic stability exists in central and western Kansas. This area corresponds to the reported range of O. virginianus texanus. In contrast, in the eastern third of the state, which corresponds to the reported range of O. virginianus macronurus, antibody prevalence is significantly lower (45%), EHDV serotypes appear to predominate, and HD, as confirmed by virus isolation, has been consistently reported. These results suggest an abrupt demarcation between enzootic stability in central and western Kansas to a pattern of epizootic HD within the eastern part of this state. Understanding host, vector, and environmental variables responsible for these contrasting patterns could have application to understanding the risk of HD in the midwestern United States. PMID- 15362830 TI - Seroprevalence of Toxoplasma gondii in Canadian pinnipeds. AB - Sera (n = 328) collected from phocids (1995-97) from the east coast of Canada, including harp seals (Phoca groenlandica), hooded seals (Cystophora cristata), grey seals (Halichoerus grypus), and harbor seals (Phoca vitulina), were diluted 1:25, 1:50, and 1:500 and tested by a modified agglutination test for antibodies to Toxoplasma gondii. Titers equal to or greater than 1:25 were considered evidence of exposure. Grey seal (11/122, 9%), harbor seal (3/34, 9%), and hooded seal (1/60, 2%) had titers of 1:25 and 1:50. Harp seals (n = 112) were seronegative. Probable maternal antibody transfer was observed in one harbor and one grey seal pup at 10 and 14 day of age, respectively. Transmission of T gondii in the marine environment is not understood. The discovery of T. gondii in marine mammals might indicate natural infections unknown because of lack of study or might indicate recent contamination of the marine environment from the terrestrial environment by natural or anthropogenic activities. PMID- 15362831 TI - Culture and serologic survey for Mycobacterium avium subsp. paratuberculosis infection among southeastern white-tailed deer (Odocoileus virginianus). AB - From July 1998 through October 2002, radiometric culture (ileocecal lymph node, mesenteric lymph node, and feces) and serologic testing by enzyme-linked immunosorbent assay (ELISA) were used to survey white-tailed deer (Odocoilens virgianus) from the soutlheastern United States for infection by Mycobacterium avium subsp. paratuberculosis (Mptb), the causative agent of paratuberculosis (Johne's disease). Mycobacterium avium subsp. paratuberculosis was isolated from the ileocecal lymph node of one of 313 deer (0.3%) originating from 63 populations in Alabama, Arkansas, Florida, Georgia, Kentucky, Louisiana, Maryland, Mississippi, North Carolina, South Carolina, Tennessee, and West Virginia (USA). Six deer (2%), all from different populations, had ELISA results above a 0.25 sample-to-positive cutoff value, but none of the ELISA reactors originated from the population from which the single Mptb isolation was made. These six deer were seronegative when tested by agar gel immunodiffusion (AGID). Collectively, these data indicate that white-tailed deer currently do not constitute a broad regional reservoir for Mptb; however, further study is warranted to clarify the significance, if any, of infected deer to the epizootiology of paratuberculosis on a local scale. Adaptation and validation of an ELISA or another serologic assay for use with deer and other wildlife would markedly enhance Mptb surveillanece among wild populations and would be a powerful tool for gaining information on the role of wild species in epidemiology of paratuberculosis. PMID- 15362832 TI - Persistence of pseudorabies virus in feral swine populations. AB - Serologic surveys for evidence of exposure to pseudorabies virus (PRV) in feral swine were conducted from November 2001 to April 2002 at 10 sites in the southeastern United States, where evidence of previous PRV exposure had been documented during 1979-89. Sera were tested in the field on the day of collection by latex agglutination. Maximum sample size per site was to be 30 animals, but sampling was discontinued before reaching this number when positive results were obtained. Positive results were obtained at all of the study sites, demonstrating long-term persistence of PRV in feral swine populations. Overall, 38 of 100 (38%) animals were positive for antibodies. Consistent results from latex agglutination tests conducted in the field and laboratory demonstrated that this test was useful as a rapid and reliable diagnostic tool when used in the field. PMID- 15362833 TI - Brucellosis in captive Rocky Mountain bighorn sheep (Ovis canadensis) caused by Brucella abortus biovar 4. AB - Nine (four female, five male) captive adult Rocky Mountain bighorn sheep (Ovis canadensis) contracted brucellosis caused by Brucella abortus biovar 4 as a result of natural exposure to an aborted elk (Cervus elaphus) fetus. Clinical signs of infection were orchitis and epididymitis in males and lymphadenitis and placentitis with abortion in females. Gross pathologic findings included enlargement of the testes or epididymides, or both, and yellow caseous abscesses and pyogranulomas of the same. Brucella abortus biovar 4 was cultured in all bighorn sheep from a variety of tissues, including testes/epididymides, mammary gland, and lymph nodes. All bighorn sheep tested were positive on a variety of standard Brucella serologic tests. This is the first report of brucellosis caused by B. abortus in Rocky Mountain bighorn sheep. It also provides evidence that bighorn sheep develop many of the manifestations ascribed to this disease and that infection can occur from natural exposure to an aborted fetus from another species. Wildlife managers responsible for bighorn sheep populations sympatric with Brucella-infected elk or bison (Bison bison) should be cognizant of the possibility of this disease in bighorn sheep. PMID- 15362834 TI - Survey for hemoparasites in imperial eagles (Aquila heliaca), steppe eagles (Aquila nipalensis), and white-tailed sea eagles (Haliaeetus albicilla) from Kazakhstan. AB - Prevalence of hemoparasites has been investigated in many avian species throughout Europe and North America. Basic hematologic surveys are the first step toward evaluating whether host-parasite prevalences observed in North America and Europe occur elsewhere in the world. We collected blood smears from 94 nestling imperial eagles (Aquila heliaca), five nestling steppe eagles (Aquila nipalensis), and 14 nestling white-tailed sea eagles (Haliaeetus albicilla) at Naurzum Zapovednik (Naurzum National Nature Reserve) in Kazakhstan during the summers of 1999 and 2000. In 1999, six of 29 imperial eagles were infected with Lencocytozoon toddi. Five of 65 imperial eagles and one of 14 white-tailed sea eagle were infected with L. toddi in 2000. Furthermore, in 2000, one of 65 imperial eagles was infected with Haemoproteus sp. We found no parasites in steppe eagles in either year, and no bird had multiple-species infections. These data are important because few hematologic studies of these eagle species have been conducted. PMID- 15362835 TI - Epidemiology of chronic wasting disease in captive white-tailed and mule deer. AB - The natural occurrence of chronic wasting disease (CWD) in a 1993 cohort of captive white-tailed deer (Odocoileus virginianus) afforded the opportunity to describe epidemic dynamics in this species and to compare dynamics with those seen in contemporary cohorts of captive mule deer (O. hemionus) also infected with CWD. The overall incidence of clinical CWD in white-tailed deer was 82% (nine of 11) among individuals that survived >15 mo. Affected white-tailed deer died or were killed because of terminal CWD at age 49-76 mo (x = 59.6 mo, SE = 3.9 mo). Epidemic dynamics of CWD in captive white-tailed deer were similar to dynamics in mule deer cohorts. Incidence of clinical CWD was 57% (4/7) among hand raised (HR) and 67% (4/6) among dam-raised (DR) mule deer; affected HR mule deer succumbed at 64-86 mo of age (x = 72 mo; SE = 5 mo), and affected DR mule deer died at age 31-58 mo (x = 41.3 mo; SE = 6.1 mo). Sustained horizontal transmission of CWD most plausibly explained epidemic dynamics, but the original source of exposures could not be determined. Apparent differences in mean age at CWD-caused death among these cohorts may be attributable to differences in the timing or intensity of exposure to CWD, and these factors appear to be more likely to influence epidemic dynamics than species differences. It follows that CWD epidemic dynamics in sympatric, free-ranging white-tailed and mule deer sharing habitats in western North American ranges also may be similar. PMID- 15362836 TI - Oral vaccination of captive arctic foxes with lyophilized SAG2 rabies vaccine. AB - Arctic foxes (Alopex lagopus) were immunized with lyophilized SAG2 oral rabies vaccine. The effectiveness of this vaccine was determined by serologic response and survival to challenge by rabies virus isolated from a red fox from Alaska (USA). No vaccine virus was found in saliva 1-72 hr after ingestion. At 2 wk after vaccination, all foxes had seroconverted, with rabies virus neutralizing antibody levels of 0.2-3.1 IU ml(-1). All vaccinated foxes survived to week 17 after challenge, and hippocampus, pons, and cerebellum were free of rabies virus as determined by direct immunofluorescence testing after death. One of four nonvaccinated foxes survived challenge and was free of rabies virus in neural tissue, and no rabies virus neutralizing antibody was detected in blood. Our results suggest that the lyophilized SAG2 oral rabies vaccine could be effective in arctic and subarctic regions, where freezing air and ground temperatures probably would not reduce its immunogenicity. PMID- 15362837 TI - West nile virus antibodies in bats from New Jersey and New York. AB - Eighty-three serum samples were obtained from big brown (Eptesicus fuscus), little brown (Myotis lucifugus), and northern long-eared (Myotis septentriotalis) bats (Chiroptera: Vespertilionidae), from New Jersey and New York (USA) between July and October 2002. Samples were analyzed for neutralizing antibodies to West Nile virus (WNV) and St. Louis encephalitis (SLE) virus. One little brown bat and one northern long-eared bat tested positive for WNV neutralizing antibodies. No bats had antibodies to SLE virus. This was the first large-scale investigation of WNV infection in bats in New Jersey. Additional work is needed to determine the effects of WNV on bat populations. PMID- 15362838 TI - Antibodies to canine distemper and phocine distemper viruses in polar bears from the Canadian arctic. AB - Serum samples collected from 200 polar bears (Ursus marititnus) from two populations in the Canadian arctic, the western Hudson Bay and Lancaster Sound populations, between 1989 and 1996, were tested for antibodies to canine distemper (CDV) and phocine distemper viruses (PDV) using virus neutralization. Antibodies to CDV and PDV were detected in 48 and six polar bears, respectively. All six bears that tested positive for PDV also tested positive for CDV; in only one case did the antibody titer for PDV exceed that of CDV. Differences in antibody prevalence to CDV were detected between populations and age classes but not sex or year of sampling. PMID- 15362839 TI - Immunogenicity of an inactivated oil-emulsion canine distemper vaccine in African wild dogs. AB - The immunogenicity of an inactivated oil-emulsion vaccine against canine distemper virus was evaluated in nine captive African wild dogs (Lycaon pictus). Antibody levels were determined by neutralization test in Vero cells. No significant local or systemic adverse reactions were observed in the animals. Virus neutralizing antibody levels >1:20 were detected, especially in animals that were vaccinated twice. The use of oil adjuvants is suggested as a good way to enhance the immune response to inactivated canine distemper vaccine. PMID- 15362840 TI - Sarcoptic mange in raccoons in Michigan. AB - Sarcoptic mange is a cause of pruritic skin disease in domestic dogs and a wide range of wildlife species. We describe sarcoptic mange in free-ranging raccoons (Procyon lotor). Three adult raccoons from upper Wayne County, Michigan (USA), were captured, killed, and submitted for diagnostic evaluation. The animals were intensely pruritic, and two had advanced alopecic and crusting lesions over their dorsum and hind limbs. Skin scrapings and skin biopsies revealed crusting and hyperkeratotic dermatitis with high numbers of Sarcoptes scabiei adults, larvae, nymphs, and eggs. These raccoons were not otherwise debilitated, with minimal internal parasites, good body condition, and no evidence of infectious bacterial or viral diseases. Because sarcoptic mange is highly contagious and affects many species, including humans, transiently, it is important that wildlife biologists and rehabilitators include sarcoptic mange in their differential list for raccoons exhibiting pruritus and alopecia. PMID- 15362841 TI - Avian pox in white-tailed laurel-pigeons from the Canary Islands. AB - Two diseased young white-tailed laurel-pigeons (Columba junoniae), an endemic and endangered species of the Canary Islands (Spain), were found in La Palma. They were very depressed and had severe cutaneous yellowish nodular lesions in feathered and unfeathered areas on the bodies of both birds. Necropsy and histopathologic analyses were conducted. The presence of epidermal hypertrophy and hyperplasia in cutaneous lesions, as well as several acidophilic intracytoplasmic inclusions in affected epithelial cells (Bollinger bodies), confirmed avian poxvirus infection. This is the first report of avian pox in whitetailed laurel-pigeons or in any other free-ranging bird in the Canaries, and it might indicate that other threatened birds of the Canarian Archipelago may be affected by this viral disease. PMID- 15362842 TI - Helminth fauna of Eurasian lynx (Lynx lynx) in Estonia. AB - Thirty-seven carcasses of Eurasian lynx (Lynx lynx) collected and examined in Estonia during 1999-2001 had helminths. Parasites identified and their prevalence included Diphyllobothrium latum (5%), Taenia pisiformis (100%), Taenia laticollis (41%), Taenia hydatigena (3%), Taenia taeniaeformis (3%), Toxocara cati (68%), and Trichinella spp. (22%). The only significant relationships (P < or = 0.05) between occurrence of helminths and host age and sex were a greater number of T. pisiformis and T. laticollis in older than in youger male lynx, and older males had a greater number of species of helminth than did younger lynx. Sixty-one fecal samples collected during snow tracking of nine lynx were examined; eggs of T. cati were identified in 38 samples, and Capillaria spp were found in eight samples. This is the first systematic investigation of parasites of lynx in Estonia. PMID- 15362843 TI - Prevalence of Escherichia coli O157:H7 in white-tailed deer from Louisiana. AB - Escherichia coli O157:H7 (EC O157) is an important zoonosis. White-tailed deer (Odocoileus virginianus) have been implicated in transmission of this bacterium to humans and have been suggested as reservoirs that might affect carriage in cattle populations. Our study objectives were to estimate prevalence of EC O157 in feces of hunter-harvested deer and to describe fecal shedding patterns in a captive herd sampled over 1 yr. Prevalence of EC O157 in hunter-harvested deer was 0.3% (n = 338). In August 2001, EC O157 was detected in one of 55 deer (1.8%) from the captive herd. Prevalence over the 1-yr period was 0.4% (n = 226). Escherichia coli O157:H7 was rarely isolated from hunter-harvested deer during the winter. We could not describe a seasonal shedding pattern based on one positive sample in the captive herd. These data do not support a prominent role of deer as a reservoir for EC O157 for cattle or humans. PMID- 15362844 TI - Pulmonary mycoplasmosis in farmed white-tailed deer (Odocoileus virginianus). AB - An outbreak of respiratory disease at a farmed cervid facility resulted in isolation and identification of Mycoplasma boris in four affected white-tailed deer (Odocoileus virginianus) fawns. Microscopically, pulmonary lesions similar to those associated with M. bovis infections in calves, inclulding lymphoplasmacytic peribronchiolar cuffing and caseonecrotic bronchiectasis, were present. Arcanobacterium pyogenes was recovered from lung tissue as well. This report indicates that M. bovis can be associated with respiratory disease in white-tailed deer. PMID- 15362845 TI - Identification of skunk species submitted for rabies testing in the desert southwest. AB - Skunks usually are identified by their common name (skunk) when submitted for rabies testing. In the desert southwest (Arizona, New Mexico, Texas, USA; and northern Mexico), there are five species of skunks; four of which can occur in sympatry. To better understand the ecology of skunk rabies in these areas, it is imperative that species be properly identified. We used the displacement loop (d loop) of the mitochondrial genome to identify to species 24 skunk brain samples submitted for rabies testing in New Mexico from 2001 to 2002. Most were identified as striped skunks (Mephitis mephitis), but hooded (Mephitis macroura) and hog-nosed (Conepatus leuconotus) skunks were also found. PMID- 15362846 TI - Unnatural amino acid mutagenesis: a precise tool for probing protein structure and function. AB - The first general method for the biosynthetic incorporation of unnatural amino acids into proteins was reported in 1989. The ensuing years have seen the solid development and subsequent implementation of "unnatural amino acid mutagenesis" in a number of groundbreaking studies. Over 100 different amino acids have been incorporated into dozens of soluble and transmembrane proteins, using both cell extract and cell-intact translation systems. The approach has provided insights into ligand-binding sites, conformational changes, and protein-protein interactions with a level of precision simply unparalleled by conventional mutagenesis. Here, the methodology is outlined, significant applications of the approach are summarized, and recent major improvements in the method are discussed. The future will likely see many more investigators utilizing this approach to manipulate proteins as it realizes its promise of becoming a tool with enormous potential. PMID- 15362847 TI - Association of spin-labeled lipids with beta-barrel proteins from the outer membrane of Escherichia coli. AB - The interaction of spin-labeled lipids with beta-barrel transmembrane proteins has been studied by the electron spin resonance (ESR) methods developed for alpha helical integral proteins. The outer membrane protein OmpA and the ferrichrome iron receptor FhuA from the outer membrane of Escherichia coli were reconstituted in bilayers of dimyristoylphosphatidylglycerol. The ESR spectra from phosphatidylglycerol spin labeled on the 14-C atom of the sn-2 chain contain a second component from motionally restricted lipids contacting the intramembranous surface of the beta-barrel, in addition to that from the fluid bilayer lipids. The stoichiometry of motionally restricted lipids, 11 and 32 lipids/monomer for OmpA and FhuA, respectively, is constant irrespective of the total lipid/protein ratio. It is proportional to the number of transmembrane beta-strands, eight for OmpA and 22 for FhuA, and correlates reasonably well with the intramembranous perimeter of the protein. Spin-labeled lipids with different polar headgroups display a differential selectivity of interaction with the two proteins. The more pronounced pattern of lipid selectivity for FhuA than for OmpA correlates with the preponderance of positively charged residues facing the lipids in the extensions of the beta-sheet and shorter interconnecting loops on the extracellular side of FhuA. PMID- 15362848 TI - Enhancing the activity of a beta-helical antifreeze protein by the engineered addition of coils. AB - The effectiveness of natural antifreeze proteins in inhibiting the growth of a seed ice crystal seems to vary with protein size. Here we have made use of the extreme regularity of the beta-helical antifreeze protein from the beetle Tenebrio molitor to explore systematically the relationship between antifreeze activity and the area of the ice-binding site. Each of the 12-amino acid, disulfide-bonded central coils of the beta-helix contains a Thr-Xaa-Thr ice binding motif. By adding coils to, and deleting coils from, the seven-coil parent antifreeze protein, we have made a series of constructs with 6-11 coils. Misfolded forms of these antifreezes were removed by ice affinity purification to accurately compare the specific activity of each construct. There was a 10-100 fold gain in activity upon going from six to nine coils, depending on the concentration that was compared. Activity was maximal for the nine-coil construct, which gave a freezing point depression of 6.5 C degrees at 0.7 mg/mL, but actually decreased for the 10- and 11-coil constructs. This small loss in activity might result from the accumulation of a slight mismatch between the spacing of the ice-binding threonine residues and the O atoms of the ice lattice. PMID- 15362849 TI - Thiamin biosynthesis in Bacillus subtilis: structure of the thiazole synthase/sulfur carrier protein complex. AB - Thiazole synthase is the key enzyme involved in the formation of the thiazole moiety of thiamin pyrophosphate. We have determined the structure of this enzyme in complex with ThiS, the sulfur carrier protein, at 3.15 A resolution. Thiazole synthase is a tetramer with 222 symmetry. The monomer is a (betaalpha)(8) barrel with similarities to the aldolase class 1 and flavin mononucleotide dependent oxidoreductase and phosphate binding superfamilies. The sulfur carrier protein (ThiS) is a compact protein with a fold similar to that of ubiquitin. The structure allowed us to model the substrate, deoxy-D-xylulose 5-phosphate (DXP), in the active site. This model identified Glu98 and Asp182 as new active site residues likely to be involved in the catalysis of thiazole formation. The function of these residues was probed by mutagenesis experiments, which confirmed that both residues are essential for thiazole formation and identified Asp182 as the base involved in the deprotonation at C3 of the thiazole synthase DXP imine. Comparison of the ThiS binding surface to the surface of ubiquitin identified a conserved hydrophobic patch of unknown function on ubiquitin that may be involved in complex formation between ubiquitin and one of its binding partners. PMID- 15362850 TI - Discovery and characterization of a substrate selective p38alpha inhibitor. AB - A novel inhibitor of p38 mitogen-activated protein kinase (p38), CMPD1, identified by high-throughput screening, is characterized herein. Unlike the p38 inhibitors described previously, this inhibitor is substrate selective and noncompetitive with ATP. In steady-state kinetics experiments, CMPD1 was observed to prevent the p38alpha-dependent phosphorylation (K(i)(app) = 330 nM) of the splice variant of mitogen-activated protein kinase-activated protein kinase 2 (MK2a) that contains a docking domain for p38alpha and p38beta, but it did not prevent the phosphorylation of ATF-2 (K(i)(app) > 20 microM). In addition to kinetic studies, isothermal titration calorimetry and surface plasmon resonance experiments were performed to elucidate the mechanism of inhibition. While isothermal titration calorimetry analysis indicated that CMPD1 binds to p38alpha, CMPD1 was not observed to compete with ATP for p38alpha, nor was it able to interrupt the binding of p38alpha to MK2a observed by surface plasmon resonance. Therefore, deuterium exchange mass spectrometry (DXMS) was employed to study the p38alpha.CMPD1 inhibitory complex, to provide new insight into the mechanism of substrate selective inhibition. The DXMS data obtained for the p38alpha.CMPD1 complex were compared to the data obtained for the p38alpha.MK2a complex and a p38alpha.active site binding inhibitor complex. Alterations in the DXMS behavior of both p38alpha and MK2a were observed upon complex formation, including but not limited to the interaction between the carboxy-terminal docking domain of MK2a and its binding groove on p38alpha. Alterations in the D(2)O exchange of p38alpha produced by CMPD1 suggest that the substrate selective inhibitor binds in the vicinity of the active site of p38alpha, resulting in perturbations to regions containing nucleotide binding pocket residues, docking groove residues (E160 and D161), and a Mg(2+) ion cofactor binding residue (D168). Although the exact mechanism of substrate selective inhibition by this novel inhibitor has not yet been disclosed, the results suggest that CMPD1 binding in the active site region of p38alpha induces perturbations that may result in the suboptimal positioning of substrates and cofactors in the transition state, resulting in selective inhibition of p38alpha activity. PMID- 15362851 TI - A duck delta1 crystallin double loop mutant provides insight into residues important for argininosuccinate lyase activity. AB - Delta-crystallin is directly related to argininosuccinate lyase (ASL), and catalyzes the reversible hydrolysis of argininosuccinate to arginine and fumarate. Two delta-crystallin isoforms exist in duck lenses, delta1 and delta2, which are 94% identical in amino acid sequence. Although the sequences of duck delta2-crystallin (ddeltac2) and duck delta1-crystallin (ddeltac1) are 69 and 71% identical to that of human ASL, respectively, only ddeltac2 has maintained ASL activity. Domain exchange experiments and comparisons of various delta-crystallin structures have suggested that the amino acid substitutions in the 20's (residues 22-31) and 70's (residues 74-89) loops of ddeltac1 are responsible for the loss of enzyme activity in this isoform. To test this hypothesis, a double loop mutant (DLM) of ddeltac1 was constructed in which all the residues that differ between the two isoforms in the 20's and 70's loops were mutated to those of ddeltac2. Contrary to expectations, kinetic analysis of the DLM found that it was enzymatically inactive. Furthermore, binding of argininosuccinate by the DLM, as well as the ddeltac1, could not be detected by isothermal titration calorimetry (ITC). To examine the conformation of the 20's and 70's loops in the DLM, and to understand why the DLM is unable to bind the substrate, its structure was determined to 2.5 A resolution. Comparison of this structure with both wild-type ddeltac1 and ddeltac2 structures reveals that the conformations of the 20's and 70's loops in the DLM mutant are very similar to those of ddeltac2. This suggests that the five amino acid substitutions in domain 1 which lie outside of the two loop regions and which are different in the DLM, and ddeltac2, must be important enzymatically. The structure of the DLM in complex with sulfate was also determined to 2.2 A resolution. This structure demonstrates that the conformational changes of the 280's loop and domain 3, previously observed in ddeltac1, also occur in the DLM upon sulfate binding, reinforcing the hypothesis that these events may occur in the active ddeltac2 protein during catalysis. PMID- 15362852 TI - Crystal structure of pyranose 2-oxidase from the white-rot fungus Peniophora sp. AB - Pyranose 2-oxidase catalyzes the oxidation of a number of carbohydrates using dioxygen. The enzyme forms a D(2) symmetric homotetramer and contains one covalently bound FAD per subunit. The structure of the enzyme from Peniophora sp. was determined by multiwavelength anomalous diffraction (MAD) based on 96 selenium sites per crystallographic asymmetric unit and subsequently refined to good-quality indices. According to its chain fold, the enzyme belongs to the large glutathione reductase family and, in a more narrow sense, to the glucose methanol-choline oxidoreductase (GMC) family. The tetramer contains a spacious central cavity from which the substrate enters one of the four active centers by penetrating a mobile barrier. Since this cavity can only be accessed by glucose sized molecules, the enzyme does not convert sugars that are part of a larger molecule. The geometry of the active center and a comparison with an inhibitor complex of the homologous enzyme cellobiose dehydrogenase allow the modeling of the reaction at a high confidence level. PMID- 15362853 TI - Influence of membrane components in the binding of proteins to membrane surfaces. AB - We have quantified the enhancement of membrane binding of activated and deactivated Galpha(s) and Galpha(q) subunits, Gbetagamma subunits, and phospholipase Cbeta(2) by lipid rafts and by the presence of membrane-associated protein partners. Membrane binding studies show that lipid rafts do not affect the intrinsic membrane affinity of Galpha(q)(GDP) and Galpha(s)(GDP), supporting the idea that these proteins partition evenly between the domains. Visualization of lipid rafts on monolayers by use of a probe that does not enter raft domains shows that neither activated nor deactivated Galpha(q)(GDP) subunits distribute evenly between the raft and nonraft domains, contrary to previous suggestions. Membrane binding of deactivated Galpha(q) and Galpha(s)(GDP) became weaker when Gbetagamma subunits were present, in contrast with the behavior predicted by thermodynamics. However, activated Galpha subunits and phospholipase Cbeta(2) were recruited to membrane surfaces by protein partners by predicted amounts. Our studies suggest that the anomalous behavior seen for deactivated Galpha subunits in the presence of Gbetagamma subunits may be due to conformational changes in the N-terminus and/or occlusion of a portion of its membrane interaction region by Gbetagamma. Even though membrane recruitment was clearly observed for one protein partner, the presence of a second partner of lower affinity did not further promote membrane binding. For these proteins, the formation of larger protein complexes with very high membrane affinities is unlikely. PMID- 15362854 TI - The canonical transient receptor potential 6 channel as a putative phosphatidylinositol 3,4,5-trisphosphate-sensitive calcium entry system. AB - We previously reported that phosphatidylinositol 3,4,5-trisphosphate (PIP(3)), a lipid product of phosphoinositide 3-kinase (PI3K), induced Ca(2+) influx via a noncapacitative pathway in platelets, Jurkat T cells, and RBL-2H3 mast cells. The identity of this Ca(2+) influx system, however, remains unclear. Here, we investigate a potential link between PIP(3)-sensitive Ca(2+) entry and the canonical transient receptor potential (TRPC) channels by developing stable human embryonic kidney (HEK) 293 cell lines expressing TRPC1, TRPC3, TRPC5, and TRPC6. Two lines of evidence support TRPC6 as a putative target by which PIP(3) induces Ca(2+) influx. First, Fura-2 fluorometric Ca(2+) analysis shows the ability of PIP(3) to selectively stimulate [Ca(2+)](i) increase in TRPC6-expressing cells. Second, pull-down analysis indicates specific interactions between biotin-PIP(3) and TRPC6 protein. Our data indicate that PIP(3) activates store-independent Ca(2+) entry in TRPC6 cells via a nonselective cation channel. Although the activating effect of PIP(3) on TRPC6 is reminiscent to that of 1-oleoyl-2-acetyl sn-glycerol, this activation is not attributable to the diacylglycerol substructure of PIP(3) since other phosphoinositides failed to trigger Ca(2+) responses. The PIP(3)-activated Ca(2+) entry is inhibited by known TRPC6 inhibitors such as Gd(3+) and SKF96365 and is independent of IP(3) production. Furthermore, we demonstrated that TRPC6 overexpression or antisense downregulation significantly alters the amplitude of PIP(3)- and anti-CD3 activated Ca(2+) responses in Jurkat T cells. Consequently, the link between TRPC6 and PIP(3)-mediated Ca(2+) entry provides a framework to account for an intimate relationship between PI3K and PLCgamma in initiating Ca(2+) response to agonist stimulation in T lymphocytes. PMID- 15362855 TI - Tyrosine 167: the origin of the radical species observed in the reaction of cytochrome c oxidase with hydrogen peroxide in Paracoccus denitrificans. AB - Determination of the three-dimensional structure of cytochrome c oxidase, the terminal enzyme of the respiratory chain, from Paracoccus denitrificans offers the possibility of site-directed mutagenesis studies to investigate the relationship between the structure and the catalytic function of the enzyme. The mechanism of electron-coupled proton transfer is still, however, poorly understood. The P(M) intermediate of the catalytic cycle is an oxoferryl state the generation of which requires one additional electron, which cannot be provided by the two metal centers. It is suggested that the missing electron is donated to this binuclear site by a tyrosine residue that forms a radical species, which can then be detected in both the P(M) and F(*) intermediates of the catalytic cycle. One possibility to produce P(M) and F(*) intermediates artificially in cytochrome c oxidase is the addition of hydrogen peroxide to the fully oxidized enzyme. Using electron paramagnetic resonance (EPR) spectroscopy, we assign a radical species detected in this reaction to a tyrosine residue. To address the question, which tyrosine residue is the origin of the radical species, several tyrosine variants of subunit I are investigated. These variants are characterized by their turnover rates, as well as using EPR and optical spectroscopy. From these experiments, it is concluded that the origin of the radical species appearing in P(M) and F(*) intermediates produced with hydrogen peroxide is tyrosine 167. The significance of this finding for the catalytic function of the enzyme is discussed. PMID- 15362856 TI - Identification and structural ramifications of a hinge domain in apolipoprotein A I discoidal high-density lipoproteins of different size. AB - Apolipoprotein (apo) A-I is the major protein constituent of human high-density lipoprotein (HDL) and is likely responsible for many of its anti-atherogenic properties. Since distinct HDL size subspecies may play different roles in interactions critical for these properties, a key question concerns how apoA-I can adjust its conformation in response to changes in HDL particle size. A prominent hypothesis states that apoA-I contains a flexible "hinge domain" that can associate/dissociate from the lipoprotein as its diameter fluctuates. Although flexible domains clearly exist within HDL-bound apoA-I, this hypothesis has not been directly tested by assessing the ability of such domains to modulate their contacts with the lipid surface. In this work, discoidal HDL particles of different size were reconstituted with a series of human apoA-I mutants containing a single reporter tryptophan residue within each of its 22 amino acid amphipathic helical repeats. The particles also contained nitroxide spin labels, potent quenchers of tryptophan fluorescence, attached to the phospholipid acyl chains. We then measured the relative exposure of each tryptophan probe with increasing quencher concentrations. We found that, although there were modest structural changes across much of apoA-I, only helices 5, 6, and 7 exhibited significant differences in terms of exposure to lipid between large (96 A) and small (78 A) HDL particles. From these results, we present a model for a putative hinge domain in the context of recent "belt" and "hairpin" models of apoA-I structure in discoidal HDL particles. PMID- 15362857 TI - Calorimetric study of the interaction of the C2 domains of classical protein kinase C isoenzymes with Ca2+ and phospholipids. AB - The affinities of Ca(2+) and anionic lipid vesicles from the C2 domains of classical protein kinase C subfamily (alpha, betaII, and gamma) were studied using isothermal titration calorimetry (ITC). In addition, the thermal stability of these C2 domains in the presence of different ligand concentrations was analyzed using differential scanning calorimetry (DSC). These three closely related C2 domains bind Ca(2+) in a similar way, demonstrating the presence of two sets of sites. The first set of sites binds one Ca(2+) ion exothermically with similar high affinity for the three proteins (K(d) around 1 microM), while the second set of sites binds endothermically approximately two Ca(2+) ions with lower affinity, which varies for each C2 domain: 22.2 microM for the PKCalpha-C2 domain, 17.2 microM for the PKCbetaII-C2 domain, and 4.3 microM for the PKCgamma C2 domain. In the absence of Ca(2+), the three C2 domains showed a weak interaction with vesicles containing anionic phospholipids. However, in the presence of a saturating Ca(2+) concentration, the C2 domains increased their affinities for the anionic lipid vesicles. In all cases, the C2 domains bound the vesicles exothermically and with similar affinities. A DSC thermal stability study of the C2 domains in the presence of Ca(2+) and anionic lipids provided further information about this protein-ligand interaction. The presence of increasing Ca(2+) concentrations was matched by an increase in the T(m) in all cases, which was even greater in the presence of anionic lipid vesicles. The extent of the change in T(m) differed for each C2 domain, reflecting the differing effect of the ligands bound during the protein stabilization. Denaturation of the C2 domains was irreversible both in the absence and in the presence of ligands, although the thermograms were not kinetically controlled. The dependence of the T(m) on the Ca(2+) concentration indicates that the protein stabilization observed by DSC primarily reflects the saturation by the cation of the low-affinity set of sites. PMID- 15362858 TI - NMR solution structure of ImB2, a protein conferring immunity to antimicrobial activity of the type IIa bacteriocin, carnobacteriocin B2. AB - Bacteriocins produced by lactic acid bacteria are potent antimicrobial compounds which are active against closely related bacteria. Producer strains are protected against the effects of their cognate bacteriocins by immunity proteins that are located on the same genetic locus and are coexpressed with the gene encoding the bacteriocin. Several structures are available for class IIa bacteriocins; however, to date, no structures are available for the corresponding immunity proteins. We report here the NMR solution structure of the 111-amino acid immunity protein for carnobacteriocin B2 (ImB2). ImB2 folds into a globular domain in aqueous solution which contains an antiparallel four-helix bundle. Extensive packing by hydrophobic side chains in adjacent helices forms the core of the protein. The C-terminus, containing a fifth helix and an extended strand, is held against the four-helix bundle by hydrophobic interactions with helices 3 and 4. Most of the charged and polar residues in the protein face the solvent. Helix 3 is well-defined to residue 55, and a stretch of nascent helix followed by an unstructured loop joins it to helix 4. No interaction is observed between ImB2 and either carnobacteriocin B2 (CbnB2) or its precursor. Protection from the action of CbnB2 is only observed when ImB2 is expressed within the cell. The loop between helices 3 and 4, and a hydrophobic pocket which it partially masks, may be important for interaction with membrane receptors responsible for sensitivity to class IIa bacteriocins. PMID- 15362859 TI - Identification and activity of a lower eukaryotic serine proteinase inhibitor (serpin) from Cyanea capillata: analysis of a jellyfish serpin, jellypin. AB - Delineating the phylogenetic relationships among members of a protein family can provide a high degree of insight into the evolution of domain structure and function relationships. To identify an early metazoan member of the high molecular weight serine proteinase inhibitor (serpin) superfamily, we initiated a cDNA library screen of the cnidarian, Cyanea capillata. We identified one serpin cDNA encoding for a full-length serpin, jellypin. Phylogenetic analysis using the deduced amino acid sequence showed that jellypin was most similar to the platyhelminthe Echinococcus multiocularis serpin and the clade P serpins, suggesting that this serpin evolved approximately 1000 million years ago (MYA). Modeling of jellypin showed that it contained all the functional elements of an inhibitory serpin. In vitro biochemical analysis confirmed that jellypin was an inhibitor of the S1 clan SA family of serine proteinases. Analysis of the interactions between the human serine proteinases, chymotrypsin, cathepsin G, and elastase, showed that jellypin inhibited these enzymes in the classical serpin manner, forming a SDS stable enzyme/inhibitor complex. These data suggest that the coevolution of serpin structure and inhibitory function date back to at least early metazoan evolution, approximately 1000 MYA. PMID- 15362860 TI - Cell-surface-associated tissue transglutaminase is a target of MMP-2 proteolysis. AB - MT1-MMP, a prototypic member of a membrane-type metalloproteinase subfamily, is an invasion promoting protease and an activator of MMP-2. In addition, MT1-MMP proteolysis regulates the functionality of cell-surface adhesion/signaling receptors including tissue transglutaminase (tTG). tTG is known to serve as an adhesion coreceptor for beta1/beta3 integrins and as an enzyme that catalyzes the cross-linking of proteins and the conjugation of polyamines to proteins. Here, we report that MMP-2, functioning in concert with MT1-MMP, hydrolyzes cell-surface associated tTG, thereby further promoting the effect initiated by the activator of MMP-2. tTG, in return, preferentially associates with the activation intermediate of MMP-2. This event decreases the rate of MMP-2 maturation and protects tTG against proteolysis by MMP-2. Our cell culture, in vitro experiments, and in silico modeling indicate that the catalytic domain of MMP-2 directly associates with the core enzymatic domain II of tTG (the K(d) = 380 nM). The follow-up cleavage of the domain II eliminates both the receptor and the enzymatic activity of tTG. Our data illuminate the coordinated interplay involving the MT1-MMP/MMP-2 protease tandem in the regulation of the cell receptors and explain the underlying biochemical mechanisms of the extensive tTG proteolysis that exists at the normal tissue/tumor boundary. Our findings also suggest that neoplasms, which express functionally active MT1-MMP and, therefore, activate soluble MMP-2, can contribute to the degradation of tTG expressed in neighboring host cells. The loss of adhesive and enzymatic activities of tTG at the interface between tumor and normal tissue will decrease cell-matrix interactions and inhibit matrix cross-linking, causing multiple pathological alterations in host cell adhesion and locomotion. PMID- 15362861 TI - Escherichia coli lipoyl synthase binds two distinct [4Fe-4S] clusters per polypeptide. AB - Lipoyl synthase (LS) is a member of a recently established class of metalloenzymes that use S-adenosyl-l-methionine (SAM) as the precursor to a high energy 5'-deoxyadenosyl 5'-radical (5'-dA(*)). In the LS reaction, the 5'-dA(*) is hypothesized to abstract hydrogen atoms from C-6 and C-8 of protein-bound octanoic acid with subsequent sulfur insertion, generating the lipoyl cofactor. Consistent with this premise, 2 equiv of SAM is required to synthesize 1 equiv of the lipoyl cofactor, and deuterium transfer from octanoyl-d(15) H-protein of the glycine cleavage system-one of the substrates for LS-has been reported [Cicchillo, R. M., Iwig, D. F., Jones, A. D., Nesbitt, N. M., Baleanu-Gogonea, C., Souder, M. G., Tu, L., and Booker, S. J. (2004) Biochemistry 43, 6378-6386]. However, the exact identity of the sulfur donor remains unknown. We report herein that LS from Escherichia coli can accommodate two [4Fe-4S] clusters per polypeptide and that this form of the enzyme is relevant to turnover. One cluster is ligated by the cysteine amino acids in the C-X(3)-C-X(2)-C motif that is common to all radical SAM enzymes, while the other is ligated by the cysteine amino acids residing in a C-X(4)-C-X(5)-C motif, which is conserved only in lipoyl synthases. When expressed in the presence of a plasmid that harbors an Azotobacter vinelandii isc operon, which is involved in Fe/S cluster biosynthesis, the as-isolated wild-type enzyme contained 6.9 +/- 0.5 irons and 6.4 +/- 0.9 sulfides per polypeptide and catalyzed formation of 0.60 equiv of 5' deoxyadenosine (5'-dA) and 0.27 equiv of lipoylated H-protein per polypeptide. The C68A-C73A-C79A triple variant, expressed and isolated under identical conditions, contained 3.0 +/- 0.1 irons and 3.6 +/- 0.4 sulfides per polypeptide, while the C94A-C98A-C101A triple variant contained 4.2 +/- 0.1 irons and 4.7 +/- 0.8 sulfides per polypeptide. Neither of these variant proteins catalyzed formation of 5'-dA or the lipoyl group. Mossbauer spectroscopy of the as-isolated wild-type protein and the two triple variants indicates that greater than 90% of all associated iron is in the configuration [4Fe-4S](2+). When wild-type LS was reconstituted with (57)Fe and sodium sulfide, it harbored considerably more iron (13.8 +/- 0.6) and sulfide (13.1 +/- 0.2) per polypeptide and catalyzed formation of 0.96 equiv of 5'-dA and 0.36 equiv of the lipoyl group. Mossbauer spectroscopy of this protein revealed that only approximately 67% +/- 6% of the iron is in the form of [4Fe-4S](2+) clusters, amounting to 9.2 +/- 0.4 irons and 8.8 +/- 0.1 sulfides or 2 [4Fe-4S](2+) clusters per polypeptide, with the remainder of the iron occurring as adventitiously bound species. Although the Mossbauer parameters of the clusters associated with each of the variants are similar, EPR spectra of the reduced forms of the cluster show small differences in spin concentration and g-values, consistent with each of these clusters as distinct species residing in each of the two cysteine-containing motifs. PMID- 15362862 TI - Only one of the two annotated Lactococcus lactis fabG genes encodes a functional beta-ketoacyl-acyl carrier protein reductase. AB - The small genome of the Gram-positive bacterium Lactococcus lactis ssp. lactis IL1403 contains two genes that encode proteins annotated as homologues of Escherichia coli beta-hydroxyacyl-acyl carrier protein (ACP) reductase. E. coli fabG encodes beta-ketoacyl-acyl carrier protein (ACP) reductase, the enzyme responsible for the first reductive step of the fatty acid synthetic cycle. Both of the L. lactis genes are adjacent to (and predicted to be cotranscribed with) other genes that encode proteins having homology to known fatty acid synthetic enzymes. Such relationships have often been used to strengthen annotations based on sequence alignments. Annotation in the case of beta-ketoacyl-ACP reductase is particularly problematic because the protein is a member of a vast protein family, the short-chain alcohol dehydrogenase/reductase (SDR) family. The recent isolation of an E. coli fabG mutant strain encoding a conditionally active beta ketoacyl-ACP reductase allowed physiological and biochemical testing of the putative L. lactishomologues. We report that expression of only one of the two L. lactis proteins (that annotated as FabG1) allows growth of the E. coli fabG strain under nonpermissive conditions and restores in vitro fatty acid synthetic ability to extracts of the mutant strain. Therefore, like E. coli, L. lactis has a single beta-ketoacyl-ACP reductase active with substrates of all fatty acid chain lengths. The second protein (annotated as FabG2), although inactive in fatty acid synthesis both in vivo and in vitro, was highly active in reduction of the model substrate, beta-ketobutyryl-CoA. As expected from work on the E. coli enzyme, the FabG1 beta-ketobutyryl-CoA reductase activity was inhibited by ACP (which blocks access to the active site) whereas the activity of FabG2 was unaffected by the presence of ACP. These results seem to be an example of a gene duplication event followed by divergence of one copy of the gene to encode a protein having a new function. PMID- 15362863 TI - Kinetic mechanism of histidine-tagged homocitrate synthase from Saccharomyces cerevisiae. AB - Kinetic data have been collected suggesting a preferred sequential ordered kinetic mechanism for the histidine-tagged homocitrate synthase (HCS) from Saccharomyces cerevisiae with alpha-ketoglutarate binding before AcCoA and CoA released before homocitrate. Oxaloacetate is also a substrate for HCS, but with lower affinity than alpha-ketoglutarate. In agreement with the ordered kinetic mechanism desulfo-CoA is uncompetitive and citrate is competitive vs alpha ketoglutarate. Varying AcCoA, citrate is a noncompetitive inhibitor as predicted, but CoA is noncompetitive vs AcCoA suggesting binding of CoA to E:homocitrate and E:alpha-ketoglutarate. The product CoA behaves in a manner identical to the dead end analogue desulfo-CoA, suggesting an E:alpha-ketoglutarate:CoA dead-end complex. Data further suggest an irreversible reaction overall, in agreement with the downhill nature of the reaction as a result of homocitryl-CoA hydrolysis. Fluorescence titration data generally agree with the steady state data, but show finite binding of CoA and AcCoA to free enzyme, suggesting that the mechanism may be random with a high degree of synergism of binding between the reactants. PMID- 15362864 TI - Calreticulin is a binding protein for muramyl dipeptide and peptidoglycan in RK13 cells. AB - Calreticulin (CRT) was isolated and identified as a protein in rabbit kidney RK(13) cells that binds the apoptogenic bacterial cell wall (BCW) components, muramyl dipeptide (MDP) and peptidoglycan (PG). Mannan-agarose purified RK(13) cell CRT (rCRT) selectively bound sepharose-immobilized L,D-MDP and PG, but not L,L-MDP or D,D-MDP. Purified rCRT and bovine CRT (bCRT) also bound free PG and L,D-MDP demonstrated in bioassays of RK(13) cell apoptosis. The results suggest that, in RK(13) cells, (a) CRT is a specific binding protein for both L,D-MDP and PG and (b) CRT binding L,D-MDP or PG is dependent on the stereoisomeric configuration of the dipeptide (L-alanyl-D-isoglutamine) moiety. In addition, the results also suggest that, in RK(13) cells, the binding of L,D-MDP, L,L-MDP, D,D MDP, or PG to CRT correlates with their capacities of inducing apoptosis. PMID- 15362865 TI - Bifunctional catalysis by CDP-ribitol synthase: convergent recruitment of reductase and cytidylyltransferase activities in Haemophilus influenzae and Staphylococcus aureus. AB - CDP-ribitol synthase catalyzes the formation of CDP-ribitol from ribulose 5 phosphate, NADPH, and CTP. CDP-ribitol is an activated precursor for the synthesis of virulence-associated polysaccharides in the capsule of the Gram negative pathogen Haemophilus influenzae and in the cell walls of Gram-positive pathogens including Staphylococcus aureus. We showed previously that CDP-ribitol synthase activity in H. influenzae is catalyzed by the bifunctional enzyme Bcs1 in a two-step reaction with reduction preceding cytidylyl transfer [Zolli, M., et al. (2001) Biochemistry 40, 5041-5048]. In the work reported here, we predicted a CDP-ribitol synthesis locus in S. aureus tandemly arranged as tarI, encoding an orthologue of the cytidylyltransferase domain of Bcs1, and tarJ, coding for an analogue of the reductase domain of Bcs1. We have shown the formation of a functional CDP-ribitol synthase complex between TarI and TarJ. Steady-state mechanistic studies of the CDP-ribitol synthases TarIJ and Bcs1 revealed that the analogous reductases and orthologous cytidylyltransferases undergo ordered mechanisms. The sequence of substrate binding and product release of the orthologous cytidylyltransferases differed. Steady-state analysis of the reductase and cytidylyltransferase activities of TarIJ indicated a 100-fold difference in the turnover where the primary reductase was rate limiting. Rapid mixing experiments revealed the presence of approximately 12 microM ribitol 5 phosphate at steady state, 100-fold lower than the observed K(m) for this intermediate. Analysis of the approach to steady state suggested that channeling was not occurring in the coupled enzyme complex and was an unlikely driving force in the convergent recruitment of reductase and cytidylyltransferase activities in the two CDP-ribitol synthases. PMID- 15362866 TI - Stability of RNA hairpin loops closed by AU base pairs. AB - Thermodynamic parameters are reported for hairpin formation in 1 M NaCl by RNA sequence of the type GCAXUAAUYUGC, where XY is the set of 10 possible mismatch base pairs. A nearest-neighbor analysis of the data indicates that the free energy of loop formation at 37 degrees C varies from 3.2 to 5.0 kcal/mol. These results combined with the model previously developed [Dale et al. (2000) RNA 6, 608] allow improvements in the model to predict the stability of RNA hairpin loops: DeltaG degrees (37L(n) = DeltaG degrees (37i(n)) + DeltaG degrees (37MM) - 0.8 (if first mismatch is GA or UU) - 0.8 (if first mismatch is GG and loop is closed on 5' side by a purine). Here, DeltaG degrees (37i(n) is the free energy for initiating a loop of n nucleotides, and DeltaG degrees (37MM) is the free energy for the interaction of the first mismatch with the closing base pair. Hairpins with GG first mismatches were found to vary in stability depending upon the orientation of the closing base pair (5' or 3' purine relative to the loop). The model gives good agreement when tested against four naturally occurring hairpin sequences. PMID- 15362867 TI - Domain topology of the DNA polymerase D complex from a hyperthermophilic archaeon Pyrococcus horikoshii. AB - Family D DNA polymerase (PolD) is a recently found DNA polymerase extensively existing in Euryarchaeota of Archaea. Here, we report the domain function of PolD in oligomerization and interaction with other proteins, which were characterized with the yeast two-hybrid (Y2H) and surface plasmon resonance (SPR) assays. A proliferating cell nuclear antigen, PhoPCNA, interacted with the N-terminus of the small subunit, DP1(1-200). Specific interaction between the remaining part of the small subunit, DP1(201-622), and the N-terminus of the large subunit, DP2(1 300), was detected by the Y2H assay. The SPR assay also indicated the intrasubunit interaction within the N-terminus, DP2(1-100), and the C-terminus, DP2(792-1163), of the large subunit. A synthetic 21 amino acid peptide corresponding to the sequence from cysteine cluster II, DP2(1290-1310), tightly interacted (a dissociation constant K(D) = 4.3 nM) with the N-terminus of the small subunit, DP1(1-200). Since the peptide could increase the 3'-5' exonuclease activity of DP1 [Shen et al. (2004) Nucleic Acids Res. 32, 158], the short region DP2(1290-1310) seems to play dual roles to form the PhoPolD complex and to regulate the 3'-5' exonuclease activity of DP1 through interaction with DP1(1 200). Furthermore, DP2(792-1163) containing the catalytic residues for DNA polymerization, Asp1122 and Asp1124, interacted with the intrasubunit domain, DP2(1-100), and the intersubunit domain, DP1(1-200). DP2(792-1163) probably forms the most important domain deeply involved in both the catalysis of DNA polymerization and stabilization of the PhoPolD complex through these multiple interactions. PMID- 15362868 TI - In vitro bypass of malondialdehyde-deoxyguanosine adducts: differential base selection during extension by the Klenow fragment of DNA polymerase I is the critical determinant of replication outcome. AB - The major malondialdehyde-derived adduct in DNA is 3-(2'-deoxy-beta-D-erythro pentofuranosyl)pyrimido[1,2-alpha]purin-10(3H)-one (M(1)dG). M(1)dG undergoes hydrolytic ring opening in duplex DNA to 9-(2'-deoxy-beta-D-erythro pentofuranosyl)-N(2)-(3-oxo-1-propenyl)guanine (N(2)OPdG). Template-primers were constructed containing M(1)dG or N(2)OPdG in a (CpG)(4) repeat sequence and replicated with the Klenow fragment of DNA polymerase I (Kf). Incorporation opposite the lesion and replication beyond the adduct sites by Kf was reduced compared to unadducted controls. The amount of bypass to full-length products was significantly greater with the acyclic adduct, N(2)OPdG, than with the cyclic adduct, M(1)dG. Sequence analysis indicated that the fully extended primers contained dC opposite both adducts when replication was conducted with Kf exo(+). In contrast, with Kf exo(-), primers extended past M(1)dG contained T opposite the adduct, but primers extended past N(2)OPdG contained dC opposite the adduct. Single nucleotide incorporation experiments indicated that Kf exo(-) incorporates all four nucleotides opposite M(1)dG or N(2)OPdG. Kf exo(+) removed dA, dG, and T opposite M(1)dG and N(2)OPdG but was much less active when dC was opposite the adduct. NMR studies on duplex DNA indicated that N(2)OPdG hydrogen bonds with dC in the complementary strand. The fact that base pairing can occur for the acyclic adduct may explain why N(2)OPdG is less blocking than M(1)dG. These results support in vivo findings that the ring-closed adduct, M(1)dG, is more mutagenic than the ring-opened adduct, N(2)OPdG. They also provide a detailed picture of in vitro replication in which the outcome is determined primarily by the selectivity of template-primer extension beyond rather than insertion opposite the adducts. PMID- 15362869 TI - Discrimination of cognate and noncognate substrates at the active site of class II lysyl-tRNA synthetase. AB - Within the two unrelated aminoacyl-tRNA synthetase classes, lysyl-tRNA synthetase (LysRS) is the only example known to exist in both classes. To probe the role of the amino acids responsible for L-lysine binding in the active site of the class II LysRS (LysRS2), we studied the lysS-encoded Escherichia coli protein. On the basis of the structure of L-lysine complexed with E. coli LysRS2 (lysS), residues implicated in amino acid recognition and discrimination were systematically replaced. Steady-state kinetic parameters for these variants showed reductions in the catalytic efficiency (k(cat)/K(M)) of 1-3 orders of magnitude, allowing the assignment of specific roles for key residues in the active site of LysRS2. To further investigate the role of each residue in discrimination against noncognate amino acids, steady-state kinetic parameters were determined for the nonprotein amino acid S-(2-aminoethyl)-L-cysteine, a potent inhibitor of LysRS2. While a number of variants showed reductions of several hundred-fold in efficiency of S (2-aminoethyl)-L-cysteine utilization, this was uniformly accompanied by similar reductions in the efficiency of lysine utilization. Thus, manipulation of the amino acid binding site only allowed up to a 4-fold improvement in S-(2 aminoethyl)-L-cysteine discrimination. This is in contrast to the highly effective discrimination against S-(2-aminoethyl)-L-cysteine by class I LysRS and correlates with the fundamentally different roles of conserved aromatic residues in the two LysRS active sites. This now provides a mechanistic basis for the proposal that differences in amino acid discrimination have been pivotal in the evolution of two unrelated LysRSs. PMID- 15362870 TI - Identification of factor XIIIA-reactive glutamine acceptor and lysine donor sites within fibronectin-binding protein (FnbA) from Staphylococcus aureus. AB - Staphylococcal fibronectin-binding protein (FnbA) is a surface-associated receptor responsible for the reversible binding of bacteria to human fibronectin and fibrin(ogen). Recently we have shown that FnbA serves as a substrate for coagulation factor XIIIa and undergoes covalent cross-linking to its ligands, resulting in the formation of heteropolymers (Matsuka, Y. V., Anderson, E. T., Milner-Fish, T., Ooi, P., and Baker, S. (2003) Staphylococcus aureus fibronectin binding protein serves as a substrate for coagulation factor XIIIa: Evidence for factor XIIIa-catalyzed covalent cross-linking to fibronectin and fibrin, Biochemistry 42, 14643-14652). Factor XIIIa also catalyzes the incorporation in FnbA of fluorescent probes dansylcadaverine and glutamine-containing synthetic peptide patterned on the NH(2)-terminal segment of fibronectin. In this study, the above probes were utilized for site-specific labeling and identification of reactive Gln and Lys residues targeted by factor XIIIa in rFnbA. Probe-decorated rFnbA samples were subjected to trypsin or Glu-C digestion, followed by separation of labeled peptides using reversed phase HPLC. Sequencing and mass spectral analyses of isolated probe-modified peptides have been employed for the identification of factor XIIIa-reactive Gln and Lys residues. Analysis of dansylcadaverine-labeled peptides resulted in the identification of one major, Gln103, and three minor, Gln105, Gln783, and Gln830, amine acceptor sites. The labeling procedure with dansyl-PGGQQIV probe revealed that Lys157, Lys503, Lys620, and Lys762 serve as amine donor sites. The identified reactive glutamine acceptor and lysine donor sites of FnbA may participate in transglutaminase mediated cross-linking reactions resulting in the covalent attachment of pathogenic Staphylococcus aureus to human host proteins. PMID- 15362871 TI - Biological activity and ferric ion binding of fragments of glycine-extended gastrin. AB - Nonamidated gastrins such as progastrin and glycine-extended gastrin17 (Ggly) induce cell proliferation and migration in vitro and colonic mucosal proliferation in vivo. Our earlier NMR study defined the structure of Ggly and showed that ferric ions are essential to its biological activity, with the first binding to Glu7 and the second to Glu8 and Glu9 (Pannequin, J. et al. (2002) J. Biol. Chem. 277, 48602-48609). The aims of this study were to define the minimum biologically active fragment of Ggly and to determine whether ferric ions were also required for its activity. Cell-proliferation studies with Ggly fragments containing the five glutamate residues showed that the nonapeptide LE(5)AYG, the octapeptide LE(5)AY, and the heptapeptides E(5)AY and LE(5)A were fully active and that their activity was dependent on the presence of ferric ions. The activity of the hexapeptides LE(5) and E(5)A and the pentapeptide E(5) was reduced and independent of the presence of iron. The stoichiometry of ferric ion binding to LE(5)AYG, LE(5)AY, and E(5)AY, determined by absorption spectroscopy, was 2 mol/mol. NMR spectroscopy showed that the nonapeptide LE(5)AYG and shorter fragments had no defined structure and that the iron-binding sites differed from those in Ggly. We conclude that, in contrast to amidated gastrins where the C terminal tetrapeptide is the minimum bioactive fragment, the shortest fully active fragments of Ggly are the heptapeptides LE(5)A and E(5)AY. These observations indicate that extensive proteolytic processing may not completely inactivate Ggly and that bioactive forms that are not detected by current radioimmunoassays may be present in tissues and/or plasma. PMID- 15362872 TI - Isomerase-independent chaperone function of cyclophilin ensures aggregation prevention of adenosine kinase both in vitro and under in vivo conditions. AB - Using inactive aggregates of adenosine kinase (AdK) from Leishmania donovani as the model substrate, we recently demonstrated that a cyclophilin (LdCyP) from the same source in an isomerase-independent fashion reactivated the enzyme in vitro by disaggregating its inactive oligomers [Chakraborty et al. (2002) J. Biol. Chem. 277, 47451-47460]. Besides disrupting preformed aggregates, LdCyP also prevents reaggregation of the newly formed active protein that is generated after productive refolding from its urea-denatured state. To investigate possible physiological implications of such phenomena, a unique expression system that simultaneously induces both AdK and LdCyP in naturally AdK-deficient Escherichia coli, was developed. Both in vitro and in vivo experiments revealed that oligomerization is an inherent property of this particular enzyme. In vivo protein cross-linking studies, activity determination analysis and Ado phosphorylation experiments carried out in cells coexpressing both the proteins unequivocally demonstrated that, similar to the phenomena observed in vitro, aggregates of the enzyme formed in vivo are able to interact with both LdCyP and its N-terminal truncated form (N(22-88)DEL LdCyP) in a crowded intracellular environment, resulting in aggregation prevention and reactivation of the enzyme. Our results indicate that the isomerase-independent chaperone function of LdCyP, detected in vitro, participates in vivo as well to keep aggregation-prone proteins in a monomeric state. Furthermore, analogous to yeast/bacterial two hybrid systems, development of this simple coexpression system may help in the confirmation of interaction of two proteins under simulated in vivo conditions. PMID- 15362873 TI - Efficient purification of recombinant proteins using hydrophobins as tags in surfactant-based two-phase systems. AB - In this work we describe the new concept of using fungal hydrophobins as efficient tags for purification of recombinant fusion proteins by aqueous two phase separation. Hydrophobins are a group of small surface-active proteins produced by filamentous fungi. Some characteristics of hydrophobins are that they are relatively small (approximately 100 amino acids), they contain eight disulfide-forming Cys residues in a conserved pattern, and they self-assemble on interfaces. The aqueous two-phase systems studied were based on nonionic surfactants that phase-separate at certain temperatures. We show that the use of hydrophobins as tags has many advantages such as high selectivity and good yield and is technically very simple to perform. Fusion proteins with target proteins of different molecular size were compared to the corresponding free proteins using a set of different surfactants. This gave an understanding on which factors influence the separation and what rationale should be used for optimization. This unusually strong and specific interaction between polymeric surfactants and a soluble protein shows promise for new developments in interfacing proteins and nonbiological materials for other applications as well. PMID- 15362874 TI - Substrate-dependent modulation of the mechanism of factor XIa inhibition. AB - Factor XIa is a serine protease which participates in both the extrinsic and intrinsic pathways of blood coagulation. In this work we used active site directed inhibitors to study the mechanism of factor IX activation by factor XIa. To this end, we developed a new sensitive method for the detection of factor IXa based on its affinity to antithrombin III. Using this assay, we found that the peptidic inhibitors, leupeptin and aprotinin, exhibited similar potencies in inhibiting factor IX activation and the cleavage of a tripeptidic chromogenic substrate by factor XIa. As expected, leupeptin and aprotinin were competitive with respect to the tripeptidic chromogenic substrate. However, the inhibition of factor IX activation was best described by mixed-type inhibition with the affinity of leupeptin and aprotinin to the factor XIa-factor IX complex only approximately 10-fold lower than their affinity toward factor XIa. These results, consistent with previous factor XI domain analyses, suggest that the active site of factor XIa does not contribute significantly to the affinity of factor XIa toward factor IX. The competitive component of the inhibition of factor IX activation suggests that binding of factor IX to factor XIa heavy chain affects the interactions of leupeptin and aprotinin with the active site. PMID- 15362875 TI - Heterogeneity in the biosynthesis of mucin O-glycans from Trypanosoma cruzi tulahuen strain with the expression of novel galactofuranosyl-containing oligosaccharides. AB - Sialoglycoprotein from Trypanosoma cruzi strains participates in important biological functions in which the O-linked glycans play a pivotal role, and their structural diversity may be related to the parasite's virulence pattern. To provide supporting evidence for this idea, we have determined the structure of novel linear and branched alpha-O-GlcNAc-linked oligosaccharides present on the mucins of the T. cruzi Tulahuen strain. The O-glycans were isolated as oligosaccharide alditols by reductive beta-elimination, purified, and characterized by nuclear magnetic resonance spectroscopy and methylation analysis. Two core families were synthesized by the parasite: the Galfbeta1- >4GlcNAc and Galpbeta1-->4GlcNAc. The Galfbeta1-->4GlcNAc core yields three series of O-chain structures. In the first, the Galf residue is nonsubstituted, while in the other series it is elongated by the activity of galactopyranosyl or galactofuranosyl transferases giving rise to Galp-beta-(1-->2)-Galf-beta-(1-->4) or Galf-beta-(1-->2)-Galf-beta-(1-->4) substructures not previously observed. The three series can arise by further galactopyranosylation of the GlcNAc O-6 arm. Sialylation was the only observed elaboration of the Galpbeta1-->4GlcNAc core family. Thus the determination of the structures of the O-glycans from T. cruzi Tulahuen mucins confirms the strain specificity of the glycosylation and predicts a relationship between it and parasite pathogenicity and the epidemiology of Chagas' disease. PMID- 15362876 TI - Apolipoproteins regulate the kinetics of endothelial lipase-mediated hydrolysis of phospholipids in reconstituted high-density lipoproteins. AB - Endothelial lipase (EL) is a newly identified member of the triglyceride lipase gene family that hydrolyzes high-density lipoprotein (HDL) phospholipids. This study investigates the ability of the major apolipoproteins of rHDL to regulate the kinetics of EL-mediated phospholipid hydrolysis in well-characterized, homogeneous preparations of spherical rHDL. The rHDL contained either apoA-I as the only apolipoprotein, (A-I)rHDL, apoA-II as the only apolipoprotein, (A II)rHDL, or apoA-I as well as apoA-II, (A-I/A-II)rHDL. The rHDL were comparable in terms of size and lipid composition and contained cholesteryl esters (CE) as their sole core lipid. Phospholipid hydrolysis was quantitated as the mass of nonesterified fatty acids (NEFA) released from the rHDL during incubation with EL. The V(max) of phospholipid hydrolysis for (A-I/A-II)rHDL [391.9 +/- 12.9 nmol of NEFA formed (mL of EL)(-1) h(-1)] was greater than (A-I)rHDL [152.8 +/- 4.7 nmol of NEFA formed (mL of EL)(-1) h(-1)]. The energy of activation (E(a)) for the hydrolysis reactions was calculated to be 52.1 and 34.8 kJ mol(-1) for (A I)rHDL and (A-I/A-II)rHDL, respectively. Minimal phospholipid hydrolysis was observed for the (A-II)rHDL. Kinetic analysis showed that EL has a higher affinity for the phospholipids in (A-I)rHDL [K(m)(app) = 0.10 +/- 0.01 mM] than in (A-I/A-II)rHDL [K(m)(app) = 0.27 +/- 0.03 mM]. Furthermore, (A-I)rHDL is a competitive inhibitor of the EL-mediated phospholipid hydrolysis of (A-I/A II)rHDL. These results establish that apolipoproteins are major determinants of the kinetics of EL-mediated phospholipid hydrolysis in rHDL. PMID- 15362877 TI - Unusually slow denaturation and refolding processes of pyrrolidone carboxyl peptidase from a hyperthermophile are highly cooperative: real-time NMR studies. AB - The refolding rate of heat-denatured cysteine-free pyrrolidone carboxyl peptidase (PCP-0SH) from Pyrococcus furiosus has been reported to be unusually slow under some conditions. To elucidate the structural basis of the unusually slow kinetics of the protein, the denaturation and refolding processes of the PCP-0SH were investigated using a real-time 2D (1)H-(15)N HSQC and CD experiments. At 2 M urea denaturation of the PCP-0SH in the acidic region, all of the native peaks in the 2D HSQC spectrum completely disappeared. The conformation of the PCP-0SH just after removal of 6 M GuHCl could be observed as a stable intermediate (D(1) state) in 2D HSQC and CD experiments, which is similar to a molten globule structure. The D(1) state of the PCP-0SH, which is the initial state of refolding, corresponded to the state at 2 M urea and seemed to be the denatured state in equilibrium with the native state under the physiological conditions. The refolding of PCP-0SH from the D(1) state to the native state could be observed to be highly cooperative without any intermediates between them, even if the refolding rate was quite slow. In the higher concentration of denaturants, PCP-0SH showed HSQC and CD spectra characteristic of completely unfolded proteins called the D(2) state. The unusually slow refolding rate was discussed as originating in the conformations in the transition state and/or the retardation of reorganization in an ensemble of nonrandom denatured structures in the D(1) state. PMID- 15362879 TI - Screening for enzyme inhibitors by surface plasmon resonance combined with mass spectrometry. AB - We have developed a novel strategy to identify enzyme inhibitors that interact directly with their enzyme targets. In the approach, an enzyme is immobilized on a sensor chip, and it is determined whether the immobilized enzyme is still active by incubation with model substrates and mass spectrometric analysis of the products. Putative inhibitors or mixtures containing putative inhibitors are then injected over the sensor chip for binding analysis with surface plasmon resonance. It is then tested whether the bound compounds inhibit the enzymatic activity by subsequent incubation with the model substrate and mass spectrometric analysis. If the bound compound inhibits the enzyme, the inhibitor is eluted from the enzyme and characterized by mass spectrometry. To test the strategy, it has been applied to the well-characterized interaction between trypsin and pure bovine pancreas trypsin inhibitor. Furthermore, fractions of plant extracts were screened for binding to and inhibition of carboxypeptidase B. PMID- 15362880 TI - Sparsely cross-linked "nanogel" matrixes as fluid, mechanically stabilized polymer networks for high-throughput microchannel DNA sequencing. AB - We have developed sparsely cross-linked "nanogels", subcolloidal polymer structures composed of covalently linked, linear polyacrylamide chains, as novel replaceable DNA sequencing matrixes for capillary and microchip electrophoresis. Nanogels were synthesized via inverse emulsion (water-in-oil) copolymerization of acrylamide and a low percentage (approximately 10(-4) mol %) of N,N-methylene bisacrylamide (Bis). Nanogels and nanogel networks were characterized by multiangle laser light scattering and rheometry, respectively, and tested for DNA sequencing in both capillaries and chips with four-color LIF detection. Typical nanogels have an average radius of approximately 230 nm, with approximately 75% of chains incorporating a Bis cross-linker. The properties and performance of nanogel matrixes are compared here to those of a linear polyacrylamide (LPA) network, matched for both polymer weight-average molar mass (M(w)) and the extent of interchain entanglements (c/c). At sequencing concentrations, the two matrixes have similar flow characteristics, important for capillary and microchip loading. However, because of the physical network stability provided by the internally cross-linked structure of the nanogels, substantially longer average read lengths are obtained under standard conditions with the nanogel matrix at a 98.5% accuracy of base-calling (for CE: 680 bases, an 18.7% improvement over LPA, with the best reads as long as 726 bases, compared to 568 bases for the LPA matrix). We further investigated the use of the nanogel matrixes in a high-throughput microfabricated DNA sequencing device consists of 96 separation channels densely fabricated on a 6-in. glass wafer. Again, preliminary DNA sequencing results show that the nanogel matrixes are capable of delivering significantly longer average read length, compared to an LPA matrix of comparable properties. Moreover, nanogel matrixes require 30% less polymer per unit volume than LPA. The addition of a small amount of low molar mass LPA or ultrahigh molar mass LPA to the optimized nanogel sequencing matrix further improves read length as well as the reproducibility of read length (RSD < 1.6%). This is the first report of a replaceable DNA sequencing matrix that provides better performance than LPA, in a side-by-side comparison of polymer matrixes appropriately matched for molar mass and the extent of interchain entanglements. These results could have significant implications for the improvement of microchip-based DNA sequencing technology. PMID- 15362881 TI - Microfluidic shear devices for quantitative analysis of cell adhesion. AB - We describe the design, construction, and characterization of microfluidic devices for studying cell adhesion and cell mechanics. The method offers multiple advantages over previous approaches, including a wide range of distractive forces, high-throughput performance, simplicity in experimental setup and control, and potential for integration with other microanalytic modules. By manipulating the geometry and surface chemistry of the microdevices, we are able to vary the shear force and the biochemistry during an experiment. The dynamics of cell detachment under different conditions can be captured simultaneously using time-lapse videomicroscopy. We demonstrate assessment of cell adhesion to fibronectin-coated substrates as a function of the shear stress or fibronectin concentration in microchannels. Furthermore, a combined perfusion-shear device is designed to maintain cell viability for long-term culture as well as to introduce exogenous reagents for biochemical studies of cell adhesion regulation. In agreement with established literature, we show that fibroblasts cultured in the combined device reduced their adhesion strength to the substrate in response to epidermal growth factor stimulation. PMID- 15362882 TI - Microfluidic T-form mixer utilizing switching electroosmotic flow. AB - This paper presents a microfluidic T-form mixer utilizing alternatively switching electroosmotic flow. The microfluidic device is fabricated on low-cost glass slides using a simple and reliable fabrication process. A switching DC field is used to generate an electroosmotic force which simultaneously drives and mixes the fluid samples. The proposed design eliminates the requirements for moving parts within the microfluidic device and delicate external control systems. Two operation modes, namely, a conventional switching mode and a novel pinched switching mode, are presented. Computer simulation is employed to predict the mixing performance attainable in both operation modes. The simulation results are then compared to those obtained experimentally. It is shown that a mixing performance as high as 97% can be achieved within a mixing distance of 1 mm downstream from the T-junction when a 60 V/cm driving voltage and a 2-Hz switching frequency are applied in the pinched switching operation mode. This study demonstrates how the driving voltage and switching frequency can be optimized to yield an enhanced mixing performance. The novel methods presented in this study provide a simple solution to mixing problems in the micro-total analysis-systems field. PMID- 15362883 TI - A three-dimensional flow control concept for single-cell experiments on a microchip. 1. Cell selection, cell retention, cell culture, cell balancing, and cell scanning. AB - An ideal microchip for single-cell experiments should be able to allow us to culture cells, to select any desired single cell from a group, to retain the cell for convenient cellular signal detection, and to deliver any buffer or reagent directly to the cell at any time during continual detection and observation. Most importantly, any negative impact on the live cell should be minimized. To accomplish all these functions, we developed a three-dimensional liquid flow control concept and employed special liquid flow fields to manipulate and retain a single yeast cell freely in the chip. A zero-speed point was controlled to retain the cell for three-dimensional cell balancing and cell scanning. A dispersive flow delivered reagents at a high speed to very near the cell and provided them to the cell at a low speed. No force stronger than its gravitational force was exerted on the cell, which could be balanced on different positions on an arc-sloping wall, thus minimizing any negative impact on the cell due to strong liquid flows. Specifically, we demonstrate on-chip single-cell culture, cell wall removal, and reagent delivery. Subsequently, single-cell fluorescence detection was performed, and noise filtering and background correction were applied for data processing. PMID- 15362884 TI - A three-dimensional flow control concept for single-cell experiments on a microchip. 2. Fluorescein diacetate metabolism and calcium mobilization in a single yeast cell as stimulated by glucose and pH changes. AB - Using a three-dimensional flow control concept to manipulate and retain a single yeast cell in a microchip, we were able to study the kinetics of intracellular metabolism and calcium mobilization at the single-cell level, as stimulated by glucose and pH changes. As a model study, the fluorogenic substrate fluorescein diacetate (FDA) was chosen to study how the intracellular carboxylesterase metabolize it. A single yeast cell was first cultured in the microchip. Thereafter, under a constant concentration of FDA, influx of FDA into the yeast cell occurred and FDA was hydrolyzed or metabolized. It was found that changes in both pH and glucose stimulated the FDA metabolism in a yeast cell, and the stimuli can elicit multiple responses from the cell. Since it was carried out within the microchip, the whole experiment on one single yeast cell could last for as long as 10 h. The dormant cell, budding cell, and pretreated budding cell (in low-pH buffer) of yeast resulted in different responses. Experimental data provided details on the FDA metabolism at the single-cell level and revealed strong correlations between FDA metabolism and calcium mobilization. Furthermore, efflux of the FDA metabolite fluorescein could start spontaneously if there was glucose in the medium. The experiments on a single cell were of the "human cell conservation" style because the cell responded to the reagent changes implemented by the human researcher. A mathematical model was also developed to study the influx-hydrolysis-efflux processes of the FDA metabolism using single-cell fluorescent data. These long overdue single-cell experiments are now rendered possible using the three-dimensional flow control in the microchip. PMID- 15362885 TI - A microfluidic system for large DNA molecule arrays. AB - Single molecule approaches offer the promise of large, exquisitely miniature ensembles for the generation of equally large data sets. Although microfluidic devices have previously been designed to manipulate single DNA molecules, many of the functionalities they embody are not applicable to very large DNA molecules, normally extracted from cells. Importantly, such microfluidic devices must work within an integrated system to enable high-throughput biological or biochemical analysis-a key measure of any device aimed at the chemical/biological interface and required if large data sets are to be created for subsequent analysis. The challenge here was to design an integrated microfluidic device to control the deposition or elongation of large DNA molecules (up to millimeters in length), which would serve as a general platform for biological/biochemical analysis to function within an integrated system that included massively parallel data collection and analysis. The approach we took was to use replica molding to construct silastic devices to consistently deposit oriented, elongated DNA molecules onto charged surfaces, creating massive single molecule arrays, which we analyzed for both physical and biochemical insights within an integrated environment that created large data sets. The overall efficacy of this approach was demonstrated by the restriction enzyme mapping and identification of single human genomic DNA molecules. PMID- 15362886 TI - Improving the signal sensitivity and photostability of DNA hybridizations on microarrays by using dye-doped core-shell silica nanoparticles. AB - The development of new highly sensitive and selective methods for microarray based analysis is a great challenge because, for many bioassays, the amount of genetic material available for analysis is extremely limited. Currently, imaging and detection of DNA microarrays are based primarily on the use of organic dyes. To overcome the problems of photobleaching and low signal intensities of organic dyes, we developed a new class of silica core-shell nanoparticles that encapsulated with cyanine dyes and applied the dye-doped nanoparticles as labeling in the DNA microarray-based bioanalysis. The developed nanoparticles have core-shell structure containing 15-nm Au colloidal cores with 95 dye alkanethiol (dT)20 oligomers chemisorbed on the each Au particle surface and 10 15-nm silica coatings bearing thiol functional groups. To be utilized for microarray detection, the dye-doped nanoparticles were conjugated with DNA signaling probes by using heterobifunctional cross-linker. The prepared nanoparticle conjugates are stable in both aqueous electrolytes and organic solvents. Two-color DNA microarray-based detection was demonstrated in this work by using Cy3- and Cy5-doped nanoparticles in sandwich hybridization. The use of the fluorophore-doped nanoparticles in high-throughput microarray detection reveals higher sensitivity with a detection limit of 1 pM for target DNA in sandwich hybridization and greater photostable signals than the direct use of organic fluorophore as labeling. A wide dynamic range of approximately 4 orders of magnitude was also found when the dye-doped nanoparticles were applied in microarray-based DNA bioanalysis. In addition, the use of these dye-doped nanoparticles as the labeling in hybridization also improved the differentiation of single-nucleotide polymorphisms. This work offers promising prospects for applying dye-doped nanoparticles as labeling for gene profiling based on DNA microarray technology. PMID- 15362887 TI - Vacuum electrolysis reactor technique for quantitation of 13-carbon isotope enrichment at the C1-position of formic acid and acetic acid. AB - A specialized vacuum electrolysis reactor was designed, constructed, and utilized for 13-carbon isotope analysis of formic acid-13C and acetic acid-13C, each highly enriched at the C1-position. This reusable reactor was equipped with two platinum wire electrodes, miniature stir bar, and sidearm reaction chamber. The associated technique developed for 13-carbon isotope analysis is based upon electrolytic generation of carbon dioxide into the preevacuated reactor followed by gas inlet mass spectrometry. It proved practical to degas and electrolyze 95% formic acid (without added electrolyte) due to adequate ionic conductivity. Formic acid-13C (nominally 99 at. % 13C) was measured by electrolytic CO2 generation to be 98.9 at. % 13C. To analyze various 13C-isotopic permutations of acetic acid, lithium and acid were separately added to reactor compartments, vacuum degassed, and stirred to produce an acidic solution. Thus, acetic acid-1 13C that was nominally 99 at. % 13C1 was determined by vacuum electrolysis to be 98.9 at. % 13C1. Further, acetic acid-2-13C that was isotope depleted at the C1 position (and known to be 99 at. % 13C at C2) gave 0.8 at. % 13C by mass spectrometry. PMID- 15362888 TI - Absolute configuration modulation attenuated total reflection ir spectroscopy: an in situ method for probing chiral recognition in liquid chromatography. AB - A method to selectively probe the different adsorption of enantiomers at chiral solid-liquid interfaces is applied, which combines attenuated total reflection infrared spectroscopy and modulation spectroscopy. The spectral changes on the surface are followed while the absolute configuration of the adsorbate is changed periodically. Demodulated spectra are calculated by performing a subsequent digital phase-sensitive data analysis. The method is sensitive solely to the difference of the interaction of the two enantiomers with the chiral surface, and the small spectral changes are amplified by the phase-sensitive data analysis. Its potential is demonstrated by investigating an already well-studied system in liquid chromatography, namely, the enantiomer separation of N-3,5-dinitrobenzoyl (R,S)-leucine (DNB-(R,S)-Leu) using tert-butylcarbamoyl quinine (tBuCQN) as the chiral selector immobilized on the surface of porous silica particles. The performed experiments and density functional theory calculations confirm an interaction model that was proposed earlier based on solution NMR and XRD in the solid state. It emerges that the ionic interaction is the strongest one, but the main reason for the potential for enantioseparation of the chiral stationary phase (CSP) is the distinct formation of a hydrogen bond of the (S)-enantiomer with the chiral selector. This H-bond is established between the amide N-H of DNB (S)-Leu with the carbamate C=O of the CSP. The (R)-enantiomer instead shows no specific hydrogen bonds. Only the unspecific ionic bonding between the protonated quinine part of the tBuCQN and the carboxylate of the DNB-(R)-Leu (holds also for DNB-(S)-Leu) is observed. PMID- 15362889 TI - Visualization of a pharmaceutical unit operation: wet granulation. AB - Recent developments in the field of process engineering and manufacturing sciences enable a new level of process understanding. However, extracting this understanding from increasing amounts of information is challenging. The aim of this study was to create a process vector from a model process describing all relevant information and, by that means, create a tool for combining and visualizing this information. Physical (impeller torque and temperature) and chemical (near-infrared spectroscopy) information from a small-scale high-shear granulation was used in the process vector. The vectors created were visualized by two different methods: principal component analysis (PCA) and the self organizing map (SOM). None of the individual measurement techniques were able to describe the state of the process alone, although they provided important information about the process. By combining the data and visualizing it, an overview could be achieved. The SOM approach had two advantages over the PCA: it presented the results in terms of the original variables and enabled the analysis of nonlinear responses. However, both visualization methods could be used to describe the progress of the process and to increase the level of process understanding. PMID- 15362890 TI - High-spatial resolution mass spectrometric imaging of peptide and protein distributions on a surface. AB - For the first time macromolecular ion microscope images have been recorded using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Single shot, mass-resolved images of the spatial distributions of intact peptide and protein ions over an area of 200 microm in diameter were obtained in less than 1 ms at a repetition rate of 12 Hz. The magnifying ion optics of the ion microscope allowed ion images to be obtained with a lateral resolution of 4 microm. These results prove the concept of high-resolution MALDI-MS imaging in microscope mode without the need for a tight laser focus and the accompanying sensitivity losses. The ion microscopy approach offers an improvement of several orders of magnitude in speed of acquisition compared to the conventional (microprobe) approach to MALDI-MS imaging. PMID- 15362891 TI - High-throughput comparative proteome analysis using a quantitative cysteinyl peptide enrichment technology. AB - A new quantitative cysteinyl-peptide enrichment technology (QCET) was developed to achieve higher efficiency, greater dynamic range, and higher throughput in quantitative proteomics that use stable-isotope labeling techniques combined with high-resolution liquid chromatography (LC)-mass spectrometry (MS). This approach involves (18)O labeling of tryptic peptides, high-efficiency enrichment of cysteine-containing peptides, and confident protein identification and quantification using the accurate mass and time tag strategy. Proteome profiling of naive and in vitro-differentiated human mammary epithelial cells using QCET resulted in the identification and quantification of 603 proteins in a single LC Fourier transform ion cyclotron resonance MS analysis. Advantages of this technology include the following: (1) a simple, highly efficient method for enriching cysteinyl-peptides; (2) a high-throughput strategy suitable for extensive proteome analysis; and (3) improved labeling efficiency for better quantitative measurements. This technology enhances both the functional analysis of biological systems and the detection of potential clinical biomarkers. PMID- 15362892 TI - Combining mass spectrometry and pull-down techniques for the study of receptor heteromerization. Direct epitope-epitope electrostatic interactions between adenosine A2A and dopamine D2 receptors. AB - Previous results from FRET and BRET experiments and computational analysis (docking simulations) have suggested that a portion of the third intracellular loop (I3) of the human dopamine D2 receptor (D2R) and the C-tail from the human adenosine A2A receptor (A2AR) are involved in A2AR-D2R heteromerization. The results of the present studies, using pull-down and mass spectrometry experiments, suggest that A2AR-D2R heteromerization depends on an electrostatic interaction between an Arg-rich epitope from the I3 of the D2R (217RRRRKR222) and two adjacent Asp residues (DD401-402) or a phosphorylated Ser (S374) residue in the C-tail of the A2AR. A GST-fusion protein containing the C-terminal domain of the A2AR (GST-A2ACT) was able to pull down the whole D2R solubilized from D2R tranfected HEK-293 cells. Second, a peptide corresponding to the Arg-rich I3 region of the D2R (215VLRRRRKRVN224) and bound to Sepharose was able to pull down both GST-A2ACT and the whole A2AR solubilized from A2AR-tranfected HEK-293 cells. Finally, mass spectometry and pull-down data showed that the Arg-rich D2R epitope binds to two different epitopes from the C-terminal part of the A2AR, containing the two adjacent Asp residues or the phosphorylated Ser residue (388HELKGVCPEPPGLDDPLAQDGAVGS412 and 370SAQEpSQGNT378). The present results are the first example of epitope-epitope electrostatic interaction underlying receptor heteromerization, a new, expanding area of protein-protein interactions. PMID- 15362893 TI - Liquid / liquid ion-transfer processes at the dioctylphosphoric acid (N,N didodecyl-N',N'-diethylphenylenediamine) / water (electrolyte) interface at graphite and mesoporous TiO2 substrates. AB - Biphasic electrode systems are studied for the case of the oxidation of the water insoluble liquid N,N-didodecyl-N',N'-diethylphenylenediamine (DDPD) neat and dissolved in bis(2-ethylhexyl) phosphate (HDOP) and immersed in aqueous electrolyte media. The oxidation process in the absence of HDOP is accompanied by transfer of the anion (perchlorate or phosphate) from the water into the organic phase. However, in the presence of HDOP, oxidation is accompanied by proton exchange instead. This electrochemically driven proton exchange process occurs over a wide pH range. Organic microdroplet deposits of DDPD in HDOP at basal plane pyrolytic graphite electrodes are studied by voltammetric techniques and compared in their behavior to organic microphase deposits in mesoporous TiO2 thin films. The mesoporous TiO2 thin film acts as a host for the organic liquid and provides an alternative biphasic electrode system compared to the random microdroplet/graphite system. Two types of mesoporous TiO2 thin-film electrodes, (i) a 300-400-nm film on ITO and (ii) a 300-400-nm film on ITO sputter-coated with a 20-nm porous gold layer, are investigated. PMID- 15362894 TI - Label-free biosensing by surface plasmon resonance of nanoparticles on glass: optimization of nanoparticle size. AB - The unique optical properties of noble metal nanoparticles have been used to design a label-free biosensor in a chip format. In this paper, we demonstrate that the size of gold nanoparticles significantly affects the sensitivity of the biosensor. Gold nanoparticles with diameters in the range of 12-48 nm were synthesized in solution and sensor chips were fabricated by chemisorption of these nanoparticles on amine-functionalized glass. Sensors fabricated from 39-nm diameter gold nanoparticles exhibited maximum sensitivity to the change of the bulk refractive index and the largest "analytical volume", defined as the region around the nanoparticle within which a change in refractive index causes a change in the optical properties of the immobilized nanoparticles. The detection limit for streptavidin-biotin binding of a sensor fabricated from 39-nm-diameter nanoparticles was 20-fold better than a previously reported sensor fabricated from 13-nm-diameter gold nanoparticles. We also discuss several other factors that could improve the performance of the next generation of these immobilized metal nanoparticle sensors. PMID- 15362895 TI - Electrogenerated chemiluminescence. 77. DNA hybridization detection at high amplification with [Ru(bpy)3]2+-containing microspheres. AB - An ultrasensitive DNA hybridization detection method based on electrogenerated chemiluminescence (ECL) using polystyrene microspheres/beads (PSB) as the carrier of the ECL labels, namely, tris(2,2'-bipyridyl)ruthenium(II) tetrakis(pentafluorophenyl)borate (Ru(bpy)3[B(C6F5)4]2), is reported. Probe single-stranded DNA (p-ssDNA) was attached to the surface of magnetic beads (MB) and hybridized with target-ssDNA (t-ssDNA) with immobilized PSB containing a large number of water insoluble Ru(bpy)3[B(C6F5)4]2 species (approximately 7.5 x 10(9) molecules/bead). With this approach a large amplification factor of Ru(bpy)3[B(C6F5)4]2 molecules for each t-ssDNA can be achieved, when each PSB is attached to a limited number of t-ssDNA. The p-ssDNA-MB <--> t-ssDNA PSB/Ru(bpy)3(2+) conjugates formed were magnetically separated from the reaction media and dissolved in MeCN containing tri-n-propylamine (TPrA) as an ECL coreactant. ECL was produced with a potential scan from 0 to 3.0 V versus Ag/Ag+, and the integrated ECL intensity was found to be linearly proportional to the t ssDNA concentration in a range of 1.0 fM to 10 nM under optimized conditions. ECL signals associated with two base pair mismatched ssDNA and noncomplementary ssDNA can be distinguished well from the ECL signal related to the complementary DNA hybridization. A Poisson distribution is followed when a large number of MB reacts with PSB, and the minimum number of 1.0- and 2.8-microm diameter MB required to bind and magnetically separate a single 10-microm diameter PSB from the reaction solution was estimated to be three and one, respectively. The principle described in this paper could be also applied to many other ECL analyses, such as immunoassays. PMID- 15362896 TI - In vitro selection of high-affinity DNA ligands for human IgE using capillary electrophoresis. AB - Aptamers with high affinity for IgE were selected using capillary electrophoresis to demonstrate the compatibility of this technique with SELEX. The high selectivity and efficiency of CE gave rise to a very high rate of enrichment, allowing high-affinity, high-selectivity aptamers to be obtained in only four rounds of selection. Decreasing the number of rounds shortens the selection procedure from the 4-6 weeks typical of SELEX to several days. The use of "bulk" dissociation constant measurements was introduced as a method for assessing the DNA pool after each round of selection. The average dissociation constant of the sequences in the DNA pool for IgE after four rounds of selection was 29 nM. The distribution of the dissociation constants for the sequences in the pool was very narrow with a standard deviation of only 6 nM. All of the sequences assessed exhibited high specificity for human IgE when compared with human IgG or mouse IgE. PMID- 15362897 TI - Capillary electrophoresis immunoassay chemiluminescence detection of zeptomoles of bone morphogenic protein-2 in rat vascular smooth muscle cells. AB - A capillary electrophoresis immunoassay (CEIA) method based on enhanced chemiluminescence (CL) detection was developed and applied to arteriosclerosis pathology research in the medical field. The system of enzyme-horseradish peroxidase (HRP) catalyzing the luminol/H2O2/p-iodophenol reaction was adopted in this paper. HRP was detected with the detection limit (S/N = 3) of 4.4 pM (53 zmol), which represents one of the highest sensitivities of HRP reported yet. HRP was first linked to bone morphogenic protein-2 (BMP-2) in rat vascular smooth muscle (VSM) cells with noncompetitive format and analyzed by CE-CL. HRP-Ab(2) mAb-BMP-2 complexes can be baseline separated from free HRP in 3 min. The detection limit (S/N = 3) of BMP-2 is 6.2 pM (75 zmol). This technique has been successfully applied to arteriosclerosis development mechanistic study by investigating the change of BMP-2 content in VSM cells, which were stimulated by angiotensin II for different times. The change trends of BMP-2 contents are well in accord with that of the commonly used pathology image analysis system. It proves that the CEIA-CL technique proposed could be developed into a sensitive and new method for clinical assay and pathology research. PMID- 15362898 TI - Electrophoresis in organogels. AB - A new matrix for electrophoresis, a low molecular weight organogel, is described. Dansylated amino acids and peptides were separated by planar and capillary electrophoresis in acetonitrile gels of trans-(1S,2S)-1,2 bis(dodecylamido)cyclohexane. The superior separation ability of the organogel over its corresponding buffer solution in capillary electrophoresis is illustrated. Organogels provide all the advantages associated with planar matrixes with 100% efficient recovery and transfer of the analytes to a mass spectrometer. We demonstrate that the planar gel can be liquefied and injected as is into an ESI-MS to identify the separands. PMID- 15362899 TI - Comparison of combination and first overtone spectral regions for near-infrared calibration models for glucose and other biomolecules in aqueous solutions. AB - Partial least squares calibration models are compared for the measurement of glucose, lactate, urea, ascorbate, triacetin, and alanine in aqueous solutions from single-beam spectra collected over the first overtone (6500-5500 cm(-1)) and the combination (5000-4000 cm(-1)) regions of the near-infrared spectrum. Spectra are collected under two sets of conditions with one designed for combination spectra and the other designed for first overtone spectra. As part of the optimization of conditions, an exponential function is presented that accurately characterizes the strong dependency between spectral quality and sample thickness. Sample thickness set for the first overtone and combination spectra are 7.5 and 1.5 mm, respectively. Independent calibration models are established for each solute from both combination and first overtone spectra. Direct comparison reveals superior performance by models generated from combination spectra, particularly for glucose and urea. Standard error of prediction (SEP) values are 1.12 and 0.45 mM for glucose models generated from first overtone and combination spectra, respectively. SEP values for urea are 7.33 and 0.10 mM for first overtone and combination spectra, respectively. Such high SEP values for urea with first overtone spectra correspond to an inability to quantify urea from these spectra because of a lack urea-specific molecular absorption features in this spectral region. Net analyte signal (NAS) is used to quantify the degree of selectivity provided within the first overtone and combination spectral regions. The superior selectivity of combination spectra is confirmed by comparing the length of the NAS vectors for each matrix component. PMID- 15362900 TI - DNA sequence detection using selective fluorescence quenching of tagged oligonucleotide probes by gold nanoparticles. AB - Simple, fast, economical, and sensitive detection of specific DNA sequences is crucial to pathogen detection and biomedical research. We have designed a novel fluorescent assay for DNA hybridization based on the electrostatic properties of DNA. We exploit the ability to create conditions where single-stranded DNA adsorbs on negatively charged gold nanoparticles while double-stranded DNA does not. Dye-tagged probe sequences have their fluorescence efficiently quenched when they are mixed with gold nanoparticles unless they hybridize with components of the analyte. Subfemtomole amounts of untagged target are detected in minutes using commercially available materials. Target sequences in complex mixtures of DNA and single-base mismatches in DNA sequences are easily detected. PMID- 15362901 TI - Improving pulse sequences for 3D diffusion-ordered NMR spectroscopy: 2DJ-IDOSY. AB - An improved pulse sequence for the 3D DOSY experiment 2DJ-DOSY, using diffusion encoding internal to the parent 2DJ spectroscopy sequence (2DJ-IDOSY), is presented. The diffusion-encoding pulses are used to enforce the desired coherence transfer pathway, reducing the minimum experimental time by at least a factor of 4, as compared to existing techniques, and approximately doubling the signal-to-noise ratio for small molecules. The new sequence is demonstrated on a simple mixture and on a complex sample with a high dynamic range (port wine). The principle of internal diffusion encoding can be applied with profit to a range of other 3D DOSY experiments. PMID- 15362902 TI - Complementary use of MALDI and ESI for the HPLC-MS/MS analysis of DNA-binding proteins. AB - Proteins from Escherichia coli were isolated based on their ability to bind DNA and digested in-solution with trypsin; the resulting peptides were separated using HPLC and subsequently analyzed using MALDI TOF/TOF and ESI Q-TOF instruments. Various properties of the peptides observed with the two ionization techniques were compared taking into account the differences between the mass analyzers. This empirical analysis of a data set containing hundreds of peptides and thousands of individual amino acids supports some of the currently held notions regarding the complementary nature of the two ionization processes. Specifically, ESI tends to favor the identification of hydrophobic peptides whereas MALDI tends to lead to the identification of basic and aromatic species. Findings from the present study suggest that ESI and MALDI may be complementary due to the biases of the two ionization techniques for certain classes of amino acids. From a practical standpoint, these biases indicate that, for the present at least, analyses must be performed on both types of instruments in order to gain the most information possible out of a given set of samples in a proteomics study. PMID- 15362903 TI - Quantitative on-line monitoring of cellular glucose and lactate metabolism in vitro with slow perfusion. AB - An on-line in vitro perfusion technique is described that allows the continuous quantification of cellular glucose metabolism in vitro. Using biosensor technology, we measure glucose and lactate metabolism at a minute-to-minute time resolution for periods up to several days. The application of our perfusion detection technique for in vitro monitoring is demonstrated in a wide variety of cells, including primary neuronal and astroglia cultures, yeast cells, and human lymphocytes. The method shows that variations in oxygen delivery or exposure to a noncompetitive pseudosubstrate (here 2-deoxyglucose) affects normal glucose metabolism. The innovative advantage of the present system is that, in contrast to other devices including a recently described system, metabolism per cell can be quantified. The potential of in vitro on-line monitoring is discussed for application in studying normal and abnormal metabolism, toxic and nontoxic drug effects, and human tissue biopsies. PMID- 15362904 TI - Development of ion drift-chemical ionization mass spectrometry. AB - An ion drift-chemical ionization mass spectrometry (ID-CIMS) technique has been developed to detect and quantify trace gases, including volatile organic compounds and inorganic species. The trace species are chemically ionized into positive or negative product ions with a well-controlled ion-molecule reaction time. The ID-CIMS method allows for quantification of the trace gases without the necessity of performing calibrations with authentic standards for the trace gases. Demonstrations of the ability of ID-CIMS to accurately quantify isoprene and HNO3 in a laboratory setting are presented. The results illustrate that the ID-CIMS technique facilitates detection and quantification of organic and inorganic species in laboratory kinetic investigations and field measurements. PMID- 15362905 TI - Use of 1.5-microm porous ethyl-bridged hybrid particles as a stationary-phase support for reversed-phase ultrahigh-pressure liquid chromatography. AB - A new ethyl-bridged hybrid packing material was evaluated in terms of its suitability for ultrahigh-pressure liquid chromatography (UHPLC). The 1.5-microm particles were obtained and packed into 30-microm-i.d. fused-silica capillary columns up to 50 cm in length. The particles were evaluated by isocratic reversed phase UHPLC at pressures up to 4500 bar (65,000 psi). The chromatographic performance of these particles was found to be similar to the performance of 1.0 microm nonporous silica particles. The mechanical strength of the ethyl-bridged hybrid material was evaluated by running a 15-cm-long column at pressures up to 4500 bar. No breakdown of the particles in the packed bed was observed. The sample loading capacity of the hybrid material was evaluated and compared to 1.0 microm nonporous silica material by observing analyte peak width versus amount injected. The observed improvement in loading capacity for the hybrid material versus nonporous silica was consistent with the improvement predicted by comparing the phase ratios of the two materials. PMID- 15362906 TI - Predictions of micelle-water partition coefficients and retention in micellar electrokinetic chromatography from solute structure. 2. Fragmental constant approach. AB - The fragmental constant approach (FCA) was used to calculate water-sodium dodecyl sulfate (SDS) micelle partition coefficients, K(mw), for uncharged solutes from their structure. Subsequently, the availability of K(mw) values allows prediction of retention factor, k, in micellar electrokinetic chromatography (MEKC) using the simple relationship k = K(mw)phi, where phi is the phase ratio. The FCA model describes a micelle-water partition coefficient as the sum of the partition coefficients of the constituent atomic/molecular fragments, measured by fragmental constant values, f (i), as well as correction factors to account for various "intramolecular effects" that cause deviations from the predicted partition coefficients as, log K(mw) = sum(n)(i=1)aif i+sum(m)(i=1)kiCm. The fragmental constants for a set of 41 fragments were determined using a training set of 229 aromatic solutes and 198 aliphatic compounds. The K(mw) of the aromatic compounds in the training set were determined by MEKC, while the K(mw) of the aliphatic solutes were estimated using the linear solvation energy relationship (LSER) for the SDS micelles. The fragments consisted of both aromatic fragments (i.e., directly attached to an aromatic ring) and aliphatic fragments. The FCA predictions agree nicely with the observed and LSER partition coefficient values, even for complex molecular structures such as beta-blocker drugs. The results show the great potential of the FCA for a priori prediction of retention behavior in MEKC from solute structure. PMID- 15362907 TI - Detector for liquid chromatography based on acoustic emissions from an oscillating flame. AB - The acoustic flame detector (AFD) is examined as a novel detector for liquid chromatography (LC). It is based upon the acoustic emission frequency of an oscillating hydrogen/oxygen premixed flame and produces a universal response toward organic molecules. A stable frequency near 1000 Hz, which further depends on mobile-phase composition, is achieved for flow rates in the microliter per minute range. The mass flow sensitivity of the AFD demonstrates a linear response over 3 orders of magnitude and a detection limit (S/sigma = 3) of approximately 15 ng of C/s for a series of alcohols. For cyclopentanol, this amounts to an injected mass of approximately 77 ng based on a 0.5-microL injection of a 196 ppm solution in methanol (flow rate 20 microL/min methanol; peak width 30 s). Similar sensitivity is observed using a water mobile phase. Low-frequency (1/f ) noise contributions are dominant with or without mobile phase present. The AFD demonstrates a uniform molar sensitivity toward carbon compounds independent of their optical properties or volatility. Results suggest the device might serve as a simple, inexpensive universal LC detector. PMID- 15362908 TI - Pinched flow fractionation: continuous size separation of particles utilizing a laminar flow profile in a pinched microchannel. AB - A concept of "pinched flow fractionation" for the continuous size separation and analysis of particles in microfabricated devices has been proposed and demonstrated. In this method, particles suspended in liquid were continuously introduced into a microchannel having a pinched segment and were aligned to one sidewall in the pinched segment by another liquid flow without particles. The particles were then separated perpendicularly to the flow direction according to their sizes by the spreading flow profile inside the microchannel. Polymer microbeads were successfully separated, and the effects of the flow rate and channel shapes on the separation performance were examined. Also, separated particles were collected independently by making branches at the end of the pinched segment. Since this method utilizes only the laminar flow profile inside a microchannel, complicated outer field control could be eliminated, which is usually required for other kinds of particle separation methods such as field flow fractionation. Also, this method can be applied both for particle size analysis and for preparation of monodispersed particles, since separation can be rapidly and continuously performed. PMID- 15362909 TI - Validation of the accuracy of the perturbation peak method for determination of multicomponent adsorption isotherm parameters in LC. AB - The isotherm parameters were for the first time determined for a quaternary mixture. This was done by the perturbation peak (PP) method using racemic mixtures of methyl and ethyl mandelate enantiomers. One complication with the PP method is that the traditional blank injection technique makes all perturbation peaks, except one, vanish at moderately nonlinear concentration plateaus. Therefore, we devised a new injection technique that made all four peaks on a quaternary component concentration plateau detectable, thereby making the determination of multicomponent competitive isotherm parameters possible. The measured quaternary perturbation data fitted well to the bi-Langmuir isotherm model and excellent agreement was found between experimental and simulated single component and multicomponent profiles, thus validating the method and the determined isotherm parameters. The method (i) is valuable for computer-assisted optimization of preparative chiral chromatography and (ii) opens the possibility of quantifying competitive drug-target interactions for chiral drugs directly on racemic mixtures, which today is impossible with any nonlabeled technique including surface plasmon resonance technology. PMID- 15362910 TI - Colloid-imprinted carbons as stationary phases for reversed-phase liquid chromatography. AB - A novel colloid-imprinting method is employed for the preparation of carbonaceous stationary phases for reversed-phase liquid chromatography (RPLC). This colloid imprinting method combined with oxidative stabilization treatment affords carbons with a porous shell/nonporous core structure. The particle morphology, pore size, pore shape, and Brunauer-Emmett-Teller surface area of these carbons can be finely tuned by selecting proper experimental conditions. Although their surface area and pore volume decrease noticeably after graphitization, their primary pore structure is maintained. In addition, the graphitization process eliminates the high-energy sites and substantially reduces structural heterogeneity, making colloid-imprinted carbons attractive stationary phases for reversed-phase liquid chromatography. The colloid-imprinted graphitic carbons with surface mesoporosity appeared to be attractive for chromatographic separations of alkylbenzenes under reversed-phase conditions. PMID- 15362911 TI - Retention indexes for temperature-programmed gas chromatography of polychlorinated biphenyls. AB - A noninteger retention index was defined based on a series of PCB internal standards, namely congeners 8 (2,4'-dichlorobiphenyl), 31 (2,4',5 trichlorobiphenyl), 44 (2,2',3,5'-tetrachlorobiphenyl), 101 (2,2',4,5,5' pentachlorobiphenyl), 138 (2,2',3,4,4',5'-hexachlorobiphenyl), 180 (2,2',3,4,4',5,5'-heptachlorobiphenyl), and 194 (2,2',3,3',4,4',5,5' octachlorobiphenyl). These retention index markers are common congeners present in technical mixtures and most environmental samples, and they show a linear dependence of retention time on the number of chlorine atoms, in the temperature programmed analysis. The index values are calculated with a single regression equation instead of the Van den Dool and Kratz equation. The retention indexes of all 209 PCBs on two commonly used columns (DB-XLB and DB-5), as well as on a supplementary column of DB-17 in capillary gas chromatography, were determined using this system. The reliability of the retention index is quite good, with the average 95% confidence limits for three measurements on each PCB being +/-0.1 index unit under the same chromatographic conditions and +/-0.4 index unit under different column head pressures. The effect of heating rate of the programmed runs on the retention index was also investigated. The inversion of the elution order of some congener pairs on the DB-XLB column for different temperature heating rates was observed. Our index values were compared with those of Castello and Testini. PMID- 15362912 TI - Preparation, characterization, and application of an enzyme-immobilized magnetic microreactor for flow injection analysis. AB - Enzyme-immobilized magnetic microparticles (EMMP) have been prepared for use as a microreactor in flow injection analysis (FI). The microparticles were directly injected into the FI system. Their retention occurred within the flow line by small permanent magnets located near the detector. The analytical utility of this concept was illustrated by the assay of glucose using glucose oxidase (GOx), immobilized microparticles, and amperometric detection of liberated hydrogen peroxide. The microparticles were derived from silica gel (nominal pore diameter, 15-80 nm) by impregnation with a citric acid/ethanol solution and a ferric nitrate/ethanol solution and then by calcination in a nitrogen atmosphere to produce ferrimagnetic fine particles of spinel-type iron oxide (gamma-Fe(2)O(3)) inside the pore. They were characterized by X-ray diffraction. The calibration curve of the glucose sample (2 microL injected) was linear between 2.5 x 10(-6) and 5 x 10(-4) mol/L (R = 0.9995), and the detection limit was 1.0 x 10(-6) mol/L or 0.36 ng of injected glucose (S/N = 3). The repeatability for a 5 x 10(-4) mol/L glucose solution was RSD = 1.5% (n = 6). Application to the assay of glucose in a fermentation broth is illustrated. The GOx MMP were stable and active for more than eight months when kept at 10 degrees C. PMID- 15362913 TI - Dialysis of persistent organic pollutants and polycyclic aromatic hydrocarbons from semipermeable membranes. A procedure using an accelerated solvent extraction device. AB - Accelerated solvent extraction (ASE) is one of the most recent solid-phase extraction methods and has caught on all over the world in numerous laboratories. Until now it was not known that this device is also very suitable for performing dialysis. In this study, development of a rapid dialysis procedure (RDP) was described that is based on the dialysis of persistent organic xenobiotics from triolein-containing semipermeable membrane devices (SPMDs) using ASE. All the operating parameters were optimized within the framework of usage. The RDP procedure was compared with the conventional dialytic recovery of target analytes under atmospheric pressure using spiked analytes and real field samples of SPMDs exposed to urban air. The main advantages of the RDP in comparison to the conventional dialysis are the speed, with up to 70 times faster taking only 40 min, and the considerable reduction in solvent consumption (by two-thirds) when SPMDs with standard configuration are used. Moreover, the RDP is also suitable as an analytical cleanup procedure for the same analytes from various types of lipid samples and other difficult matrixes using semipermeable membranes. PMID- 15362914 TI - Direct determination of lead isotopes (206Pb, 207Pb, 208Pb) in arctic ice samples at picogram per gram levels using inductively coupled plasma-sector field MS coupled with a high-efficiency sample introduction system. AB - Adopting strict cleanroom procedures, ice samples from the Canadian High Arctic have been analyzed for Pb concentrations and Pb isotopes (206Pb, 207Pb, 208Pb) using ICP-SMS. The detection limit for Pb (0.06 pg g(-1)) was approximately 2 orders of magnitude lower than the lowest concentration of Pb in the ice samples (range, 4.3-1660 pg g(-1); median, 45 pg g(-1)). Acidification of ice samples with high-purity HNO3 for stabilization purposes contributed only 0.004 pg of Pb g(-1), which is an insignificant source of Pb. Using a new sample introduction system consisting of a heated (140 degrees C) minicyclonic spray chamber and a Peltier cooled condenser (2 degrees C) and by replacing the conventional sample cone with a high-performance cone, signal intensities for Pb were increased by approximately 1 order of magnitude. Thus, it was possible not only to measure Pb isotope ratios directly using ICP-SMS but also to achieve reasonable precision (approximately 0.2%) at low picogram per gram concentrations of total Pb. This precision is comparable to that achievable by thermal ionization mass spectrometry at such low Pb concentrations, but the ICP-SMS requires much less sample volume (approximately 2 mL), needs no sample pretreatment, and therefore is considerably faster and less expensive than the conventional approach. Even though absolute Pb concentrations in two ice samples dating from 1974 and 1852 were very similar (9 and 6 pg g(-1)) their fundamentally different isotopic signature (206Pb/207Pb: 1.169 +/- 0.002 vs 1.147 +/- 0.003) clearly indicates different sources of Pb. The analytical procedures described here, therefore, offer great promise for fingerprinting the predominant sources of atmospheric Pb in polar snow and ice. PMID- 15362915 TI - Determination of perchlorate anion in foods by ion chromatography-tandem mass spectrometry. AB - A rapid, sensitive, and specific method was developed for determining perchlorate anion in lettuce, cantaloupe, bottled water, and milk. A test portion of chopped crop homogenate was extracted with diluted nitric acid and filtered. Milk proteins were precipitated with acetonitrile, and the supernatant, after centrifugation, was cleaned up on a graphitized carbon solid-phase extraction column. Water samples were analyzed directly. All test solutions were syringe filtered and mixed with an 18O4-labeled perchlorate internal standard before ion chromatography-tandem mass spectrometry. A strong anion exchange column eluted with 100 mM ammonium acetate in 50:50 (v/v) acetonitrile/water was interfaced via electrospray ionization to a triple stage quadrupole mass spectrometer operated in the negative ion mode. The labeled internal standard corrected for any sample matrix effects on measured signals. Four parent-to-product ion transitions, for loss of oxygen, were monitored for native and 18O4-labeled perchlorate anion, respectively: 35Cl-perchlorate, m/z 99 --> 83 and 107 --> 89; 37Cl-perchlorate, m/z 101 --> 85 and 109 --> 91. The limit of quantitation was 1.0 microg/kg in lettuce, 2.0 microg/kg in cantaloupe, 0.50 microg/L in bottled water, and 3.0 microg/L in milk. Native perchlorate was recovered from fortified test portions in the range 93-107% for lettuce, 107-114% for cantaloupe, 100-115% for bottled water, and 99-101% for milk. PMID- 15362916 TI - Discovery and neurochemical screening of peptides in brain extracellular fluid by chemical analysis of in vivo microdialysis samples. AB - Endogenous peptides from brain extracellular fluid of live rats were analyzed using capillary liquid chromatography (LC)-tandem mass spectrometry (MS2). A 4-mm long microdialysis probe perfused at 0.6 microL/min implanted into the striatum of anesthetized male rats was used to collect 3.6 microL dialysate fractions that were injected on-line into the capillary LC-MS2 system for analysis. A total of 3349 MS2 spectra were collected from 13 different animals under basal conditions and during localized depolarization evoked by infusion of a high-K+ solution through the microdialysis probe. Subtractive analysis revealed a total of 859 MS2 spectra that were observed only during depolarization. From these spectra, 29 peptide sequences (25 were peptides not previously observed) from 6 different protein precursors were identified using database searching software. Proteins identified include precursors to neuropeptides, synaptic proteins, blood proteins, and transporters. The identified peptides represent candidates for neurotransmitters, neuromodulators, and markers of synaptic activity or brain tissue damage. A screen for neuroactivity of novel proenkephalin fragments that were found was performed by infusing the peptides into the brain while monitoring amino acid neurotransmitters by microdialysis sampling combined with capillary electrophoresis. Three of the six tested peptides evoked significant increases in various neuroactive amino acids. These results demonstrate that this combination of methods can identify novel neurotransmitter candidates and screen for potential neuroactivity. PMID- 15362917 TI - Selective multiphoton ionization of coplanar polychlorobiphenyls using 266-nm laser emission by gas chromatography/mass spectrometry. AB - Polychlorinated biphenyls (PCBs), used as a heat exchange oil (Kanechlor, KC 300), can be measured by gas chromatography combined with multiphoton ionization mass spectrometry (GC/MPI-MS). Several compounds, e.g., 4,4'-dichlorobiphenyl and 3,4,4'-trichlorobiphenyl, have nearly the same retention time but can be selectively determined by MPI-MS. Coplanar PCBs are more efficiently ionized using the fourth harmonic emission of a Nd:YAG laser (266 nm), compared to noncoplanar PCBs. Thus, the approach reported herein may be useful in the selective as well as the sensitive analysis of toxic PCBs, contained in old transformers and capacitors that have been mandated by the government to be properly disposed of within 10 years in Japan. PMID- 15362918 TI - Corona discharge neutralizer for electrospray aerosols used with condensation nucleation light-scattering detection. AB - To obviate the use of radioactive materials, a simple unipolar electrical neutralizer based on a corona discharge was developed to neutralize electrospray aerosols used when coupling capillary liquid chromatography (CLC) and capillary electrophoresis (CE) to condensation nucleation light-scattering detection (CNLSD). The electrical neutralizer could be operated with either a positive or negative corona to generate ions to neutralize oppositely charged aerosols generated by electrospray and allow sensitive detection with CNLSD. The device could further be operated with organic solvents as well as aqueous buffer. The parameters affecting the corona discharge and neutralization process were studied and optimized. Compared to a CNLSD system using a radioactive neutralizer, the system with the electrical neutralizer showed higher signal and signal-to-noise ratio for test compounds, which indicated better neutralization efficiency of the charged aerosol. Highly reproducible results were obtained with the robust and durable electrical neutralizer with both CLC- and CE-electrospray-CNLSD systems. PMID- 15362919 TI - Amperometric glucose biosensor based on assisted ion transfer through gel supported microinterfaces. AB - A novel amperometric glucose sensor was developed based on the facilitated proton transfer across microinterfaces between two immiscible electrolyte solutions. The combination of a 1,3:2,4-dibenzylidene sorbitol/2-nitrophenyl octyl ether gel membrane and 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine as the ionophore allows the transfer of protons from water to the gellified organic phase; the gel membrane is supported on arrays of microholes drilled on a polyester film. The protons are generated as the result of the dissociation of gluconic acid produced during the enzymatic degradation of glucose by glucose oxidase. The characteristics of the glucose sensor were investigated using several experimental conditions, namely, the concentration of ligand and enzyme. The electrochemical response is typical of an enzymatic electrode and displays a linear behavior in the range 0.2-3 mM glucose. The effect of the experimental parameters of the voltammetric technique was also optimized with the aim of improving sensor sensitivity. PMID- 15362920 TI - Gallium nitride-based potentiometric anion sensor. AB - The gallium nitride (GaN) semiconductor is used as the sensing element for the development of a potentiometric anion sensor. The anion recognition mechanism is based on the selective interaction of anions in solution with the epitaxial Ga face polarity GaN (0001) wurtzite crystal film grown on sapphire. The native GaN crystal is used for the development of an ion blocked sensor. The potential is based on the Volta potential, generated at the semiconductor/solution interface and within the Helmholtz layer, due to specifically adsorbed anions. The selectivity of the sensor is based on the direct interaction of the anionic ligand with the outer electron-defective gallium atoms; thus, it is not dependent on the lipophilicity of the adsorbed charged species. The chemical resistivity of the GaN crystal provides sensors with excellent lifetime, signal stability, and reproducibility. PMID- 15362921 TI - Voltammetric sensor for oxidized DNA using ultrathin films of osmium and ruthenium metallopolymers. AB - Films containing [Os(bpy)2(PVP)10Cl]+ and [Ru(bpy)2(PVP)10Cl]+ metallopolymers were assembled layer by layer on pyrolytic graphite electrodes to make sensors that selectively detect oxidized DNA. These films showed reversible, independent electrochemistry for electroactive Os3+/Os2+ and Ru3+/Ru2+ centers, with formal potentials of 0.34 and 0.76 V vs SCE, respectively. The combination of ruthenium and osmium metallopolymers in the films provided a catalytic Os square wave voltammetry (SWV) peak that is mainly selective for 8-oxoguanine and the detection of other oxidized nucleobases from the Ru peak. The method is applicable to measurements on DNA in solution or DNA incorporated into films. Using the Os SWV peak, 1 oxidized nucleobase in 6000 was detected. The sensor is simple and inexpensive, and the approach may be useful for the detection of oxidized DNA as a clinical biomarker for oxidative stress. PMID- 15362922 TI - In situ surface-enhanced infrared study of hydrogen bond pairing of complementary nucleic acid bases at the electrochemical interface. AB - Surface-enhanced infrared absorption spectroscopy with a Kretschmann-type attenuated total reflection configuration has been used to study hydrogen-bonded pairing between 6-amino-8-purinethiol, a thiol-derivatized adenine, immobilized on a gold electrode surface, and thymidine, a complimentary base derivative of adenine, in 0.1 M NaClO4 aqueous solution as a function of applied potential. 6 Amino-8-purinethiol is adsorbed on a gold surface via a sulfur atom to form a S Au bond. Nearly half of the adsorbed molecules are protonated, and the long axis of the adenine moiety is tilted from the surface normal at open circuit potential. As the potential increases, the acid-base equilibrium is shifted toward the unprotonated form and the adenine moiety is reoriented toward a nearly perpendicular configuration. The hydrogen bond interaction between the adsorbed 6 amino-8-purinethiol with thymidine in solution is greatly affected by the protonation and orientation of the adenine moiety and is controllable by the applied potential. Due to steric hindrance, an adenine-thymine-type hydrogen bond pair is formed only at potentials more positive than 0.1 V (vs SCE) where the unprotonated adenine moiety is perpendicularly oriented. PMID- 15362923 TI - Chronoamperometry at micropipet electrodes for determination of diffusion coefficients and transferred charges at liquid/liquid interfaces. AB - Chronoamperometry was carried out at liquid/liquid interfaces supported at the tip of micropipet electrodes for direct determination of the diffusion coefficient of a species in the outer solution. The diffusion coefficient was used for subsequent determination of the transferred charges per species from the diffusion-limited steady-state current. A large tip resistance of the micropipets causes prolonged charging current so that the faradic current can be measured accurately only at a long-time regime (typically t > 5 ms). At the same time, the long-time current response at the interfaces surrounded by a thin glass wall of the pipets is enhanced by diffusion of the species from behind the pipet tip. Therefore, numerical simulations of the long-time chronoamperometric response were carried out using the finite element method for accurate determination of diffusion coefficients. Validity of the simulation results was confirmed by studying simple transfer of tetraethylammonium ion. The technique was applied for transfer/adsorption reactions of the natural polypeptide protamine and also for Ca2+ and Mg2+ transfers facilitated by ionophore ETH 129. With the diffusion coefficient of protamine determined to be (1.2 +/- 0.1) x 10(-6) cm(2)/s, the ionic charge transferred by each protamine molecule was obtained as +20 +/- 1, which is close to the excess positive charge of protamine. Also, the diffusion coefficient of ETH 129 was determined to demonstrate that each ionophore molecule transfers +0.67 and +1 charge per Ca2+ and Mg2+ transfer, respectively, which corresponds to formation of 1:3 and 1:2 complexes with the respective ions. PMID- 15362924 TI - Numerical investigation of transient current density distributions for multi-ion electrolytes at a rotating disk electrode. AB - A versatile model for the simulation of transient multiion transport and reaction processes is applied to investigate current density distributions over a rotating disk electrode for linear voltammetric sweep experiments. The model accounts for ion transport by convection, diffusion, and migration, in combination with Butler Volmer type electrode reactions. For several process conditions (reversible and irreversible reactions, excess or lack of supporting electrolyte), the current density distribution over the disk surface is examined and the transient current response is compared to results from the more commonly used one-dimensional axial approach. The impact of migrational effects on the nonuniform local process conditions over the disk surface is illustrated, and the resulting effect on the current peak height, width, and position is investigated. A mathematical correlation for the current peak height as a function of the reacting ion transference number is established. PMID- 15362925 TI - Solvent bar microextraction. AB - In this work, a new and simple microextraction method termed solvent bar microextraction (SBME) was developed. In this method, the organic extractant solvent (1-octanol) was confined within a short length of a hollow fiber membrane (sealed at both ends) that was placed in a stirred aqueous sample solution. Tumbling of the extraction device within the sample solution facilitated extraction. Pentachlorobenzene (PCB) and hexachlorobenzene (HCB) were used as model compounds to investigate the extraction performance. Analysis was carried out by gas chromatography/electron capture detection. This new method provided very high enrichment (approximately 110-fold for PCB and approximately 70-fold for HCB) in 10 min and good reproducibility (<4%, n = 6). Since the hollow fiber membrane was sealed, it could be used for extraction from "dirty" samples, such soil slurries. This novel microextraction method was compared with single-drop microextraction and static hollow fiber membrane microextraction in which the extractant solvent was also held within a hollow fiber but with the latter fixed to a syringe needle (i.e., there was no tumbling effect). Comparison between SBME and conventional solid-phase microextraction in a soil slurry sample was also investigated. PMID- 15362926 TI - Bonding of glass microfluidic chips at room temperatures. AB - A simple, room-temperature bonding process was developed for the fabrication of glass microfluidic chips. High-quality bonding with high yields (>95%) was achieved without the requirement of clean room facilities, programmed high temperature furnaces, pressurized water sources, adhesives, or pressurizing weights. The plates to be bonded were sequentially prewashed with acetone, detergent, high-flow-rate (10-20 m/s) tap water, and absolute ethyl alcohol and were soaked in concentrated sulfuric acid for 8-12 h. The plates were again washed in high-flow-rate tap water for 5 min and, finally, with demineralized water. The plates were bonded by bringing the cleaned surfaces into close contact under a continuous flow of demineralized water and air-dried at room temperature for more than 3 h. This bonding process features simple operation, good smoothness of the plate surface, and high bonding yield. The procedures can be readily applied in any routine laboratory. The bonding strength of glass chips thus produced, measured using a shear force testing procedure, was higher than 6 kg/cm(2). The mechanism for the strong bonding strength is presumably related to the formation of a hydrolyzed layer on the plate surfaces after soaking the substrates in acid or water for extended periods. Microfluidic chips bonded by the above procedure were tested in the CE separation of fluorescein isothiocyanate-labeled amino acids. PMID- 15362927 TI - Pharmacoeconomic implications of new therapies in sepsis. AB - Severe sepsis is a major healthcare problem, characterised by a high incidence, mortality and cost. New breakthroughs in treatment are quite diverse, including: (i) more effective regimens for generic, inexpensive broad anti-inflammatory agents (corticosteroids); (ii) a recombinant protein (drotrecogin-alfa [activated]); and (iii) a protocol-based treatment approach (early goal-directed therapy). Economic analyses of new sepsis agents should adopt the societal perspective, which requires prolonging the time horizon beyond that currently typically studied in sepsis trials, so that patient-centred outcomes can be more fully captured. Sepsis affects a very diverse group of patients, and if findings are to be generalisable, careful attention must be paid to study entry criteria and differences in effects and costs across different patient subgroups. Existing care patterns for sepsis are also quite diverse, with the consequence that the incremental effects on costs and outcomes could vary widely by practice pattern, again affecting generalisability. Furthermore, many sepsis patients receive multiple other therapies, which together with therapies under study may have varied and unintended, potentially costly or dangerous adverse effects, which could have a large influence on cost-effectiveness estimates. Finally, there are a number of large yet potentially hidden costs of sepsis, such as the long-term costs of managing patients who develop sepsis or the costs of introducing different interventions into clinical practice. Such costs must also be addressed in economic analyses. The search for new anti-sepsis strategies remains vigorous and exciting. We recommend wider incorporation of economic analyses into the study of potential new therapies, with appropriate attention to the caveats discussed above. Clinical demand to use new agents must be balanced against the economic consequences of their use. PMID- 15362928 TI - Pharmacoeconomic burden of undertreating hypertension. AB - Many studies have shown the importance of antihypertensive drug therapy as a factor in reducing the risk of cardiovascular morbidity and mortality, and in containing the cost of managing hypertension and its complications. Nevertheless, the evidence in clinical practice indicates about half of hypertensive patients do not receive pharmacological treatment and about half of treated patients do not achieve blood pressure level control. Undertreating hypertension is the leading cause of failure in drug therapy effectiveness and cost effectiveness. The pharmacoeconomic burden of undertreating hypertension can be defined as the clinical (number of cardiovascular events) and economic (costs of managing cardiovascular events) consequences that would have been avoided by adequate control of blood pressure levels. In the last few years, the increase in this burden and the restriction of budget constraints has raised the awareness of healthcare providers with regards to the need to achieve better performance and to improve disease management of hypertension. This review aims to present the current situation regarding the pharmacoeconomic burden of undertreating hypertension by identifying the key issues of this medical condition, defining and measuring the extent of undertreatment, defining and measuring costs associated with undertreatment, and discussing some fundamental aspects of disease management for hypertension. The pharmacoeconomic burden of undertreating hypertension appears to be an extremely important phenomenon for which there is currently only very limited adequate research. The present dearth of appropriate data can be largely attributed to the lack of epidemiological studies in clinical practice. Future studies are necessary for a more precise quantification of the therapeutic and economic impact of undertreating arterial hypertension in clinical practice (appropriateness studies) and for more precise selection of antihypertensive drugs on the basis of the different cost-effectiveness profiles detected in 'real world' settings (cost-effectiveness studies). PMID- 15362929 TI - Cost-benefit analysis of first-generation antihistamines in the treatment of allergic rhinitis. AB - BACKGROUND: The majority of individuals with allergic rhinitis in the US take first-generation antihistamines (FGAs). Although FGAs have been proven effective in alleviating allergic rhinitis symptoms, they have been associated with an increased risk of motor vehicle, aviation and occupational injuries and deaths, reduced productivity and impaired learning. OBJECTIVE: The objective of this analysis was to quantify the total costs and benefits of FGA use in the US from the societal perspective. METHODS: We used a decision-analytic model to quantify the annual societal costs and benefits of treatment with FGAs compared with the hypothetical alternative of no treatment for the population of individuals with allergic rhinitis and taking FGAs in the US in 2001. The benefit associated with FGA use was estimated using the willingness-to-pay framework and projected to the US population using published estimates of the prevalence of allergic rhinitis. The costs of FGA-associated sedation included lost productivity and the direct and indirect cost of unintentional injuries (including motor vehicle, occupational, public and home injuries and fatalities). The incidence of injuries and fatalities associated with FGA use was estimated using the risk of injury attributable to the sedentary effects of FGAs in the allergic rhinitis population. To evaluate uncertainty in the model assumptions, a probabilistic sensitivity analysis was conducted using Bayesian second-order Monte Carlo simulation. Costs and benefits are expressed in 2001 US dollars, using a 3% discount rate. RESULTS: Based on current utilisation, the total societal benefit (95% credible interval) associated with the use of FGAs for the treatment of allergic rhinitis was US 7.7 billion dollars (US 1.3 billion dollars to US 21 billion dollars). The societal cost of purchasing FGAs was only US 697 million dollars. However, the societal cost of FGA-associated sedation was US 11.3 billion dollars (US 2.4 billion dollars to US 50.8 billion dollars). The annual societal net benefit of FGA use for the treatment of allergic rhinitis in the US was -US4.2 billion dollars (-US 36 billion dollars to +US 0.296 billion dollars). The net benefit was negative in 97% of the 10,000 Monte Carlo simulations. CONCLUSIONS: The societal benefits of FGA use in alleviating the symptoms of allergic rhinitis are significant. However, based on the assumptions, probability distributions and parameter estimate ranges used in the current model, it is very likely that the costs associated with sedation exceed the benefits of FGA use in the US. The cost of FGA-associated sedation is comparable to estimates of the cost of all medical care expenditures on respiratory conditions in the US (US 12.1 billion dollars to US 31.3 billion dollars) [1996 values] and provides compelling evidence of the economic burden of sedation associated with FGA use. PMID- 15362930 TI - The costs and benefits of community thrombolysis for acute myocardial infarction : a decision-analytic model. AB - BACKGROUND: There is evidence that the earlier a patient reaches hospital and receives thrombolysis, the better the outcome. The GREAT (Grampian Region Early Anistreplase Trial) directly addressed the issue of early thrombolysis by evaluating, in a randomised controlled trial, the efficacy of thrombolysis in the community compared with that administered in hospital. OBJECTIVE: This paper aimed to model the cost and benefits of community compared with hospital thrombolysis from the UK NHS perspective, using efficacy data from the GREAT. METHODS: A decision-analytic approach was used to model these two alternatives. Resource use and cost estimates were estimated for a single tertiary centre. Estimates of effectiveness in life-years were obtained from the 4-year follow-up for patients recruited to the GREAT, using declining exponential approximation of life expectancy. Costs are in pounds sterling, 2000/1 values. RESULTS: Community thrombolysis had an average life expectancy of 12.48 years and hospital thrombolysis had an average life expectancy of 12.39 years. Costs were 361 pounds sterling for community thrombolysis and 300 pounds sterling for hospital thrombolysis. Community thrombolysis led to an additional 0.09 years of life expectancy gained compared with hospital thrombolysis at an additional cost of 61 pounds sterling per patient. Therefore, the incremental cost per life-year gained for the community thrombolysis service over the hospital thrombolysis service was 667 pounds sterling. Sensitivity analysis showed that estimates of cost per life year gained were most sensitive to the estimates of survival. CONCLUSION: This model suggests that, from the UK NHS perspective, implementing community thrombolysis may lead to extra survival but at extra cost over hospital thrombolysis. Although the incremental cost per life-year is modest, judgements still have to be made, however, as to whether the extra benefits estimated are worth the additional resources required. This requires consideration of the local context in which the service may be introduced. PMID- 15362931 TI - Ground- and excited-state proton transfer reaction of 3-hydroxyflavone in dimyristoylphosphatidylcholine liposome membrane. AB - 3-Hydroxyflavone (3HF), a molecule that exhibits excited-state intramolecular proton transfer, has been studied for its fluorescence characteristics in dimyristoylphosphatidylcholine (DMPC) liposome membrane. 3HF partitions to the lipid bilayer membrane with a reasonably large partition coefficient. On excitation at 417 nm, a weak emission from the ground-state anion species was observed at 483 nm, whereas excitation at absorption maxima (345 nm) gives the usual intense fluorescence of the phototautomeric emission at 530 nm. In this article, we report the observation of a ground-state proton transfer reaction of 3HF in DMPC liposome membrane. PMID- 15362932 TI - Bioluminescence imaging of the response of rat gliosarcoma to ALA-PpIX-mediated photodynamic therapy. AB - Photodynamic therapy (PDT) is a promising modality for the treatment of solid tumors that combines a photosensitizing agent and light to produce cytotoxic reactive oxygen species that lead to tumor cell death. The recent introduction of bioluminescence imaging (BLI), involving the use of the luciferase gene (luc) transferred into target tumor cells, followed by systemic administration of luciferin and detection of the emitted visible chemiluminescence photons, offers the potential for longitudinal imaging of tumor growth and therapeutic response in single animals. We demonstrate in this study the first results of the use of BLI to assess the response of an intracranial brain tumor model (9L rat gliosarcoma) to aminolevulinic acid (ALA)-mediated PDT. Complementary in vitro experiments with the luciferase-transfected 9L cells show that the decrease in the luminescent signal after PDT correlates with cell kill. In vivo imaging shows a decrease in the BLI signal from the tumor after ALA-PDT treatment, followed by tumor regrowth. Furthermore, preliminary studies using cells transfected with a hypoxia-responsive vector show an increase in bioluminescence within 4 h after Photofrin-mediated PDT, demonstrating the ability to observe stress-gene responses. These results suggest that BLI can be used to provide spatiotemporal information of intracranial brain tumor responses after PDT and may serve as a valuable response-endpoint measure. PMID- 15362934 TI - Influence of eicosapentaenoic acid, an omega-3 fatty acid, on ultraviolet-B generation of prostaglandin-E2 and proinflammatory cytokines interleukin-1 beta, tumor necrosis factor-alpha, interleukin-6 and interleukin-8 in human skin in vivo. AB - Dietary omega-3 polyunsaturated fatty acids (omega-3 PUFA) reduce sunburn, an acute inflammatory response, in humans. We assessed whether this may be mediated by reduced ultraviolet-B (UV-B) induction of proinflammatory mediators tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, IL-6, IL-8 and prostaglandin (PG)E(2) in healthy skin. In a double-blind, randomized study, 28 humans received 4 g daily of 95% ethyl esters of eicosapentaenoic acid (EPA) or oleic acid (OA) orally for 3 months. Skin biopsies and suction blister fluid were taken from unexposed and UV-B-exposed skin and examined for mediator expression immunohistochemically and quantitatively by immunoassay; plasma levels were also assayed. The subjects taking EPA, but not OA, showed a significant rise in their minimal erythemal dose (MED) (data reported elsewhere). Before supplementation, irradiation with 3x MED UV-B increased blister fluid TNF-alpha, IL-6, IL-8 and PGE(2) at 16 h (all P < 0.001). No significant change occurred in baseline or UV B-induced skin levels of cytokines after either supplement, whereas UV-B induction of PGE(2) was abolished after EPA but not OA. Immunohistochemical expression of the cytokines at baseline and after UV-B was unaltered by EPA and OA; circulating cytokine and PGE(2) levels were also unchanged. Hence, in healthy skin in vivo, there was no evidence that reduction of the sunburn response by EPA is mediated by the proinflammatory cytokines examined; abrogation of UV-B generated PGE(2) may play a role. PMID- 15362935 TI - Involvement of human small fragment nuclease in the resistance of human cells to UV-C-induced cell death. AB - Human small fragment nuclease (Sfn) is one of the cellular proteins that were reported to degrade small, single-stranded DNA and RNA. However, the biological role of Sfn in cellular response to various stressors such as UV-C (mainly 254 nm wavelength ultraviolet ray) remains unclear. We have examined whether modulation of human SFN gene expression affects cell survival capacity against UV-C-induced cell death, analyzing colony survival ability in UV-C-sensitive human RSa cells treated with short double-stranded RNA (siRNA) specific for SFN messenger RNA (mRNA). The expression levels of SFN mRNA in the siRNA-treated RSa cells decreased to about 15% compared with those in the control siRNA-treated cells. The siRNA-treated RSa cells showed lower colony survival and higher activity of caspase-3 after UV-C irradiation than the control siRNA-treated RSa cells. Furthermore, the removal capacity of cyclobutane pyrimidine dimers (CPD) in the siRNA-treated RSa cells decreased compared with the control siRNA-treated RSa cells. There was no difference in the colony survival and CPD removal capacity after UV-C irradiation between the control siRNA-treated RSa cells and mock treated RSa cells. These results suggest that SFN expression is involved in resistance of RSa cells to UV-C-induced cell death through the roles it plays in the DNA repair process. PMID- 15362936 TI - Energy transfer and fluorescence quenching in complexes of polymethine dyes with human serum albumin. AB - Electronic excitation energy transfer (EET) between molecules of polymethine dyes bound to human serum albumin (HSA) has been established and studied by absorption and fluorescence spectroscopy as well as by fluorescence decay measurements. In this system, excitation of the donor dye molecule leads to fluorescence of the acceptor dye molecule, both bound to HSA, with donor fluorescence quenching by the acceptor. The short distance between the donor and the acceptor (25-28 A) revealed from the Forster model of EET as well as some spectroscopic data show that both molecules are probably located in the same binding domain of HSA. The role of HSA is to bring donor and acceptor molecules together to a distance adequate to achieve EET as well as to increase the donor and acceptor fluorescence quantum yields. Efficient quenching of the intrinsic HSA fluorescence by some polymethine dyes (oxonols) is observed. The experimental results fit well a model for the formation of a weakly fluorescent dye-HSA complex; the quencher in this complex should be located in the immediate vicinity of the HSA fluorophore group (Trp(214)). PMID- 15362937 TI - UV absorption and photoisomerization of p-methoxycinnamate grafted silicone. AB - p-Methoxycinnamate moieties, UV-B-absorptive chromophores of the widely used UV-B filter, 2-ethylhexyl p-methoxycinnamate (OMC), were grafted onto the 7 mol% amino functionalized silicone polymer through amide linkages. Comparing with OMC, the resulting poly [3-(p-methoxycinnamido)(propyl)(methyl)-dimethyl] siloxane copolymer (CAS) showed less E to Z isomerization when exposed to UV-B light. The absorption profiles of the product showed the maximum absorption wavelength to be similar to that of OMC but with less sensitivity to the type of solvent. Poly (methylhydrosiloxane) grafted with 10 mol% p-methoxycinnamoyl moieties was prepared through hydrosilylations of 2-propenyl-p-methoxycinnamate, in which the resulting copolymer showed similar results to those of CAS. PMID- 15362939 TI - The effect of molecular environment on the photoisomerization of urocanic acid. AB - Urocanic acid, imidazole propenoic acid, is a metabolic product of histidine, which accumulates in skin and is excreted in sweat. It absorbs UV radiation at wavelengths shorter than 340 nm, and its principal photochemical reaction is a trans-cis isomerization about the propenyl double bond. This isomerization to the biologically active cis isomer is implicated in the photoinduced suppression of the immune system of skin. The kinetics of the trans --> cis photoisomerization of urocanic acid has been determined in a number of solvents, spanning a range of polarities. The initial rates of isomerization and the photostationary trans-cis compositions, in all solvents except water, correlate linearly with solvent polarity. This indicates that the isomerization proceeds through a polar intermediate that is stabilized by coulombic interactions with the molecular environment. PMID- 15362940 TI - Radical cation generation from singlet and triplet excited states of all-trans lycopene in chloroform. AB - On direct photoexcitation, subpicosecond time-resolved absorption spectroscopy revealed that the 1B(u)-type singlet excited state of all-trans-lycopene in chloroform was about seven times more efficient than all-trans-beta-carotene in generating the radical cation. The time constant of radical cation generation from the 1B(u)-type state was found to be approximately 0.14 ps, a value that was comparable for the two carotenoids. On anthracene-sensitized triplet excitation, radical cation generation was found to be much less efficient for lycopene than for beta-carotene. A slow rising phase (20-30 micros) in the bleaching of ground state absorption was common for both lycopene and beta-carotene in chloroform and was ascribed to an efficient secondary reaction with a solvent radical leading to the formation of carotenoid radical cations. The reverse ordering in the tendency of the excited states of different multiplicities for the two carotenoids to generate radical cations is discussed in relation to the two carotenoids as scavengers of free radicals. PMID- 15362942 TI - Photodecomposition of Pigment Yellow 74, a pigment used in tattoo inks. AB - Tattooing has become a popular recreational practice among younger adults over the past decade. Although some of the pigments used in tattooing have been described, very little is known concerning the toxicology, phototoxicology or photochemistry of these pigments. Seven yellow tattoo inks were obtained from commercial sources and their pigments extracted, identified and quantitatively analyzed. The monoazo compound Pigment Yellow 74 (PY74; CI 11741) was found to be the major pigment in several of the tattoo inks. Solutions of commercial PY74 in tetrahydrofuran (THF) were deoxygenated using argon gas, and the photochemical reaction products were determined after exposure to simulated solar light generated by a filtered 6.5 kW xenon arc lamp. Spectrophotometric and high pressure liquid chromatography (HPLC) analyses indicated that PY74 photodecomposed to multiple products that were isolated using a combination of silica chromatography and reversed-phase HPLC. Three of the major photodecomposition products were identified by nuclear magnetic resonance and mass spectrometry as N-(2-methoxyphenyl)-3-oxobutanamide (o-acetoacetanisidide), 2-(hydroxyimine)-N-(2-methoxyphenyl)-3-oxobutanamide and N,N''-bis(2 methoxyphenyl)urea. These results demonstrate that PY74 is not photostable in THF and that photochemical lysis occurs at several sites in PY74 including the hydrazone and amide groups. The data also suggest that the use of PY74 in tattoo inks could potentially result in the formation of photolysis products, resulting in toxicity at the tattoo site after irradiation with sunlight or more intense light sources. PMID- 15362943 TI - Photoinactivation of hepatitis A virus by synthetic porphyrins. AB - Porphyrins are photosensitizers and may be applicable in situations where viral inactivation is required, as for in vitro inactivation of nonenveloped viruses in blood components or in other aqueous media. No study has examined the efficacy of porphyrin inactivation on human pathogens such as hepatitis A virus (HAV) in plasma or other liquids. Experiments were conducted to evaluate the effect of synthetic porphyrins on HAV in porphyrin-containing human plasma and phosphate buffered saline exposed to long-wavelength (365 nm) UV light. Inactivation of bacteriophage MS2 (MS2) also was determined in some trials. Solutions containing cationic, anionic or amphiphilic porphyrins irradiated with an average light dose of 4.3 J/cm(2) for 90 min resulted in >3 log(10) (>99.9%) to >4 log(10) (>99.99%) inactivation of both HAV and MS2. Viral inactivation may have been greater than observed because the limits of detection of the assay had been reached. Under ambient lighting conditions, none of the porphyrins was mutagenic in the Ames assay and only the congener with the longest chain-length, tetrakis (N-[n hexadecyl]-4-pyridiniumyl) porphyrin, was appreciably toxic to mammalian cells. Disinfection by photoactivated synthetic porphyrins therefore can offer an effective and relatively safe approach to removal of nonenveloped viruses from aqueous media. PMID- 15362944 TI - Plant responses to current solar ultraviolet-B radiation and to supplemented solar ultraviolet-B radiation simulating ozone depletion: an experimental comparison. AB - Field experiments assessing UV-B effects on plants have been conducted using two contrasting techniques: supplementation of solar UV-B with radiation from fluorescent UV lamps and the exclusion of solar UV-B with filters. We compared these two approaches by growing lettuce and oat simultaneously under three conditions: UV-B exclusion, near-ambient UV-B (control) and UV-B supplementation (simulating a 30% ozone depletion). This permitted computation of "solar UV-B" and "supplemental UV-B" effects. Microclimate and photosynthetically active radiation were the same under the two treatments and the control. Excluding UV-B changed total UV-B radiation more than did supplementing UV-B, but the UV-B supplementation contained more "biologically effective" shortwave radiation. For oat, solar UV-B had a greater effect than supplemental UV-B on main shoot leaf area and main shoot mass, but supplemental UV-B had a greater effect on leaf and tiller number and UV-B-absorbing compounds. For lettuce, growth and stomatal density generally responded similarly to both solar UV-B and supplemented UV-B radiation, but UV-absorbing compounds responded more to supplemental UV-B, as in oat. Because of the marked spectral differences between the techniques, experiments using UV-B exclusion are most suited to assessing effects of present day UV-B radiation, whereas UV-B supplementation experiments are most appropriate for addressing the ozone depletion issue. PMID- 15362945 TI - Simultaneous production of superoxide radical and singlet oxygen by sulphonated chloroaluminum phthalocyanine incorporated in human low-density lipoproteins: implications for photodynamic therapy. AB - Sulfonated chloroaluminum phthalocyanines have been studied for their use in the photodynamic therapy (PDT) of tumors. Plasma low-density lipoproteins (LDL) are important carriers of phthalocyanines in the blood, but on exposure to visible light, phthalocyanine-loaded LDL undergo an oxidation process that propagates to erythrocytes. We attempted to identify the reactive species involved in LDL and erythrocyte oxidation by means of electron paramagnetic resonance (EPR) spectroscopy in the presence of 2,2,6,6-tetramethyl-4-piperidone (TEMP) and the spin trap 5,5'-dimethyl-1-pyrroline-N-oxide (DMPO). Irradiation of phthalocyanine loaded LDL in the presence of DMPO resulted in the formation of a four-line EPR spectrum with relative intensity of 1:2:2:1 (a(N) = a(H) = 14.8 G), characteristic of DMPO-hydroxyl radical spin adduct. This signal was sensitive to superoxide dismutase and slightly sensitive to catalase, but a mixture of the two enzymatic activities was the most efficient in promoting a decrease in the intensity of the EPR signal. In the presence of erythrocytes, an increase in the quartet intensity for a hematocrit of 1% and 4% was observed, decreasing for higher erythrocyte concentrations. The irradiation of phthalocyanine-loaded LDL in the presence of TEMP resulted in the formation of a nitroxide radical, 2,2,6,6 tetramethyl-4-piperidone-N-oxyl radical, intensity of which was sensitive to histidine, a singlet oxygen ((1)O(2)) quencher. Under both incubation conditions, with DMPO and TEMP, the formation of the respective EPR signals required the sensitizer (phthalocyanine), light and oxygen. Overall, the results are compatible with the simultaneous formation of superoxide anion and (1)O(2), implying that Type-I and Type-II mechanisms of photochemistry are simultaneously operative in phthalocyanine-loaded LDL. However, for a constant LDL/phthalocyanine ratio, the formation of oxygen free radicals shows a biphasic behavior with the concentration of LDL increasing and reaching a plateau, whereas the formation of (1)O(2) increases linearly with LDL concentration. Erythrocytes at high (physiological) concentrations induced a decrease in the intensity of both EPR signals. The physiological relevance of these findings in the framework of PDT is briefly discussed. PMID- 15362946 TI - A novel mitochondrial signaling pathway activated by visible-to-near infrared radiation. AB - The number of cells attached to glass substratum increases if HeLa cell suspension is irradiated with monochromatic visible-to-near infrared radiation before plating (the action spectrum with maxima at 619, 657, 675, 700, 740, 760, 800, 820, 840 and 860 nm). Treating of cell suspension with sodium azide (2 x 10( 5) M), sodium nitroprusside (5 x 10(-5) M), ouabain (1 x 10(-6) M) or amiloride (1.7 x 10(-5) M) before irradiation significantly modifies the spectrum of cell attachment enhancement. A light-induced mitochondrial signaling pathway can be regulated by small ligands directly binding to the catalytic center of cytochrome c oxidase (N(3), NO) as well as by chemicals specifically binding to plasma membrane enzymes (ouabain, amiloride). The comparative analysis of action spectra allows the conclusions that first, Cu(A) and Cu(B) chromophores of cytochrome c oxidase could be involved as photoacceptors and second, various signaling pathways (reaction channels) between cytochrome c oxidase and cell attachment regulation are at work. PMID- 15362947 TI - Comparative sensitivity of microvascular endothelial cells, fibroblasts and tumor cells after in vitro photodynamic therapy with meso-tetra-hydroxyphenyl-chlorin. AB - The phototoxic effect of meso-tetra-hydroxyphenyl-chlorin (mTHPC)-mediated photodynamic therapy (PDT) on human microvascular endothelial cells (hMVEC) was compared with that on human fibroblasts (BCT-27) and two human tumor cell lines (HMESO-1 and HNXOE). To examine the relationship between intrinsic phototoxicity and intracellular mTHPC content, we expressed cell survival as a function of cellular fluorescence. On the basis of total cell fluorescence, HNXOE tumor cells were the most sensitive and BCT-27 fibroblasts the most resistant, but these differences disappeared after correcting for cell volume. Endothelial cells were not intrinsically more sensitive to mTHPC-PDT than tumor cells or fibroblasts. Uptake of mTHPC in hMVEC increased linearly to at least 48 h, whereas drug uptake in the other cell lines reached a maximum by 24 h. No difference in drug uptake was seen between the cell lines during the first 24 h, but by 48 h hMVEC had a 1.8- to 2.8-fold higher uptake than other cell lines. Endothelial cells showed a rapid apoptotic response after mTHPC-mediated PDT, whereas similar protocols gave a delayed apoptotic or necrotic like response in HNXOE. We conclude that endothelial cells are not intrinsically more sensitive than other cell types to mTHPC-mediated PDT but that continued drug uptake beyond 24 h may lead to higher intracellular drug levels and increased photosensitivity under certain conditions. PMID- 15362949 TI - Estimating UV erythemal irradiance by means of neural networks. AB - In recent years, there has been a substantial increase in attempts to model the flux of ultraviolet radiation (UV). UV irradiance at surface level is a result of the combined effects of solar zenith angle, surface elevation, cloud cover, aerosol load and optical properties, surface albedo and the vertical profile of ozone. In this study, we present the development of an artificial neural network (ANN) model that can be used to estimate solar UV irradiance on the basis of optical air mass, ozone columnar content, latitude, horizontal visibility data and cloud information such as type, coverage and height. ANN are widely accepted as a technology offering an alternative way to tackle complex and ill-defined problems. They can learn from examples, are fault tolerant in the sense that they are able to handle noisy and incomplete data, are able to deal with nonlinear problems and, once trained, can perform prediction and generalization at high speed. In this study, a multilayer perceptron network (MLP) consisting of an input layer, an output layer and one hidden layer was used. Training of the neural network was done using the Bayesian regulation back propagation algorithm. The study was developed using data from three stations on the Iberian Peninsula: Madrid and Murcia during the period 2000-2001 and Zaragoza in 2001. To train and validate the MPL neural networks, independent subsets of data were extracted from the complete database at each station. The results suggest that a MLP neural network using optical air mass, ozone columnar content, latitude and total cloud coverage provides the best estimates, with mean bias deviation and root mean square deviation of -0.1% and 18.0%, 1.6% and 19.6%, 0.1% and 14.6% at Madrid, Murcia and Zaragoza, respectively. Despite the dependence of the cloud radiative effect on cloud type, the use of additional information such as cloud type or cloud elevation did not improve these results. The performance of the developed ANN has been checked regarding its ability to estimate the UV index (UVI); results indicate that in more than 95% of the cases, the difference between estimated and measured values does not exceed one unit of UVI. PMID- 15362950 TI - Lifetime of fluorescent pyrene butyric acid probe in single living cells for measurement of oxygen fluctuation. AB - We study the fluorescence lifetime of the well-known 1-pyrene butyric acid (PBA) to assess oxygen concentrations in living cells. The behavior of the probe is first studied in water, ethanol, protein solution and liposome suspension. The Stern-Volmer plot of these solutions is linear, and the bimolecular reaction rate constant agrees with previous observations. In single living cells, the PBA lifetime decreases with oxygen concentration (185 to 55 ns). The probe lifetime differences between living cells and liposome suspension, especially under nitrogen atmosphere, suggest a supplemental pathway for the deactivation of the probe. We simplify further the complex living cells system by stopping the cell functions and studying freshly fixed cells. In this case, we obtained an increase of PBA lifetime under nitrogen atmosphere (215 ns). PMID- 15362951 TI - Photochemical behavior of a new long-chain UV absorber derived from 4-tert-butyl 4'-methoxydibenzoylmethane. AB - A new UV filter, the 1-(4-tert-butylphenyl)-2-decanyl-3-(4'-methoxyphenyl) propane-1,3-dione, called C10-DBM, was prepared by grafting a 10-carbon aliphatic chain to the alpha-carbonyl position of 4-tert-butyl-4'-methoxydibenzoylmethane (BM-DBM), a well-known and often used UV filter. The UV-A absorption efficiency of organic solutions containing the new filter was tested and compared with identical solutions containing BM-DBM with or without irradiation (xenon lamp). The originality of this new filter is that its UV-A absorbance appeared during irradiation of the molecule. Although the molar absorption coefficient of C10-DBM in the UV-A domain was lower than that of BM-DBM, the solutions absorption exhibited a much more photostable behavior under irradiation. In this study, we first demonstrated that C10-DBM was a precursor of BM-DBM (enol isomer) by means of high-performance liquid chromatography followed by mass spectrometry. Indeed, we showed that the UV-A absorption of C10-DBM solutions appearing during the irradiation of the molecule was due to a Norrish-II reaction (beta-cleavage), which induced the release of the BM-DBM enol form and 1-decene. Then, we established a kinetic model for the photochemistry of C10-DBM and fitted the variation of UV absorption spectra to confirm the proposed mechanism. PMID- 15362952 TI - Photodynamic studies and photoinactivation of Escherichia coli using meso substituted cationic porphyrin derivatives with asymmetric charge distribution. AB - The photodynamic activities of novel asymmetrically meso-substituted cationic porphyrins, 5,10-di(4-methylphenyl)-15,20-di(4-trimethylammoniumphenyl)porphyrin iodide 1 and 5-(4-trifluorophenyl)-10,15,20-tris(4 trimethylammoniumphenyl)porphyrin iodide 2 and its metal complex with Pd(II) 3, have been investigated in both homogeneous medium bearing photooxidizable substrates and in vitro on a typical gram-negative bacterium Escherichia coli. The amphiphilic character of porphyrin 2 was increased by the presence of a high lipophilic trifluoromethyl group and its photophysical properties changed by forming a complex with Pd(II). Absorption and fluorescence spectroscopic studies were compared in different media. Fluorescence quantum yields (phi(F)) of 0.16 for 1 in tetrahydrofuran and 0.08 for 2 in N, N-dimethylformamide (DMF) were calculated, whereas no significant emission was detected for Pd(II) porphyrin 3. The singlet molecular oxygen, O(2)((1)Delta(g)), production was evaluated using 9,10-dimethylanthracene in DMF yielding relative values of 1, 0.55 and 0.47 for porphyrins 3, 2 and 1, respectively. A faster decomposition of l-tryptophan was obtained using Pd(II) porphyrin 3 as sensitizer with respect to the free-base porphyrins 1 and 2. In biological medium, the behavior of cationic porphyrins 1-3 were compared with that of 5-(4-carboxyphenyl)-10,15,20-tris(4 methylphenyl)porphyrin 4, which was used as a noncationic sensitizer. These porphyrins are rapidly bound to E. coli cells in 5 min and the amount of cell bound sensitizer is not appreciably changed incubating the cultures for longer times. The recovered porphyrin 2 after one washing step reaches a value of approximately 2.9 nmol/10(6) cells and this amount remains high even after three washes, indicating that this sensitizer is tightly bound to cells. Photosensitized inactivation of E. coli was analyzed using cells without and with one washing step. In both cases, a higher photoinactivation of cells was found for tricationic porphyrin 2 and 3, causing a approximately 5.5 log (99.999%) decrease of cell survival, when treated with 10 microM of sensitizer. Under these conditions, a lower effect was found for porphyrin 1 (approximately 4 log) whereas sensitizer 4 did not produce appreciable photodamage. The results were also confirmed by growth delay experiments. These studies show that the amphiphilic tricationic porphyrin 2 and 3 bearing a trifluoromethyl group can be a promising model for phototherapeutic agents with potential applications in inactivation of bacteria by photodynamic therapy. PMID- 15362953 TI - Shedding some light on tattoos? PMID- 15362954 TI - Arginine-482 is not essential for transport of antibiotics, primary bile acids and unconjugated sterols by the human breast cancer resistance protein (ABCG2). AB - The human BCRP (breast cancer resistance protein, also known as ABCG2) is an ABC (ATP-binding cassette) transporter that extrudes various anticancer drugs from cells, causing multidrug resistance. To study the molecular determinants of drug specificity of BCRP in more detail, we have expressed wild-type BCRP (BCRP-R) and the drug-selected cancer cell line-associated R482G (Arg482-->Gly) mutant BCRP (BCRP-G) in Lactococcus lactis. Drug resistance and the rate of drug efflux in BCRP-expressing cells were proportional to the expression level of the protein and affected by the R482G mutation, pointing to a direct role of BCRP in drug transport in L. lactis. In agreement with observations in mammalian cells, the BCRP-R-mediated transport of the cationic substrates rhodamine 123 and tetramethylrosamine was significantly decreased compared with the activity of BCRP-G. In addition, BCRP-R showed an enhanced interaction with the anionic anticancer drug methotrexate when compared with BCRP-G, suggesting that structure/substrate specificity relationships in BCRP, as observed in eukaryotic expression systems, are maintained in prokaryotic L. lactis. Interestingly, BCRP R exhibited a previously unestablished ability to transport antibiotics, unconjugated sterols and primary bile acids in L. lactis, for which the R482G mutation was not critical. Since Arg482 is predicted to be present in the intracellular domain of BCRP, close to transmembrane segment 3, our results point to a role of this residue in electrostatic interactions with charged substrates including rhodamine 123 and methotrexate. Since unconjugated sterols are neutral molecules and bile acids and many antibiotics are engaged in protonation/deprotonation equilibria at physiological pH, our observations may point either to a lack of interaction between Arg482 and neutral or neutralized moieties in these substrates during transport or to the interaction of these substrates with regions in BCRP not including Arg482. PMID- 15362955 TI - Real-time imaging of myosin II regulatory light-chain phosphorylation using a new protein biosensor. AB - Phosphorylation of the RMLC (regulatory myosin light chain) regulates the activity of myosin II, which is critically involved in the motility of both muscle and non-muscle cells. There are both Ca2+-dependent and -independent pathways for RMLC phosphorylation in smooth-muscle cells, and the latter pathway is often involved in an abnormal contractility in pathological states such as asthma and hypertension. Therefore pharmacological interventions of RMLC phosphorylation may have a therapeutic value. In the present study, we developed a new genetically encoded biosensor, termed CRCit (ECFP-RMLC-Citrine, where ECFP is enhanced cyan fluorescent protein), that detects RMLC phosphorylation using fluorescence resonance energy transfer between two variants of the green fluorescent protein fused to both the N- and C-termini of RMLC. When expressed in primary cultured vascular smooth-muscle cells, CRCit detected the Ca2+-dependent RMLC phosphorylation with a high spatiotemporal resolution. Furthermore, we could specifically assay the agonist-induced Ca2+-independent phosphorylation of RMLC when Ca2+ signalling in cells expressing CRCit was suppressed. Thus CRCit may also be used for the high throughput screening of compounds that inhibit abnormal smooth-muscle contraction. PMID- 15362956 TI - Transcriptional regulation of the human DNA methyltransferase 3A and 3B genes by Sp3 and Sp1 zinc finger proteins. AB - The DNMT3A (DNA methyltransferase 3A) and DNMT3B genes encode putative de novo methyltransferases and show complex transcriptional regulation in the presence of three and two different promoters respectively. All promoters of DNMT3A and DNMT3B lack typical TATA sequences adjacent to their transcription start sites and contain several Sp1-binding sites. The importance of these Sp1-binding sites was demonstrated by using a GC-rich DNA-binding protein inhibitor, mithramycin A, i.e. on the basis of decrease in the promoter activities and mRNA expression levels of DNMT3A and DNMT3B. Overexpression of Sp1 and Sp3 up-regulated the promoter activities of these two genes. The physical binding of Sp1 and Sp3 to DNMT3A and DNMT3B promoters was confirmed by a gel shift assay. Interestingly, Sp3 overexpression in HEK-293T cells (human embryonic kidney 293T cells) resulted in 3.3- and 4.0-fold increase in DNMT3A and DNMT3B mRNA expression levels respectively by quantitative reverse transcriptase-PCR, whereas Sp1 overexpression did not. Furthermore, an antisense oligonucleotide to Sp3 significantly decreased the mRNA levels of DNMT3A and DNMT3B. These results indicate the functional importance of Sp proteins, particularly Sp3, in the regulation of DNMT3A and DNMT3B gene expression. PMID- 15362957 TI - Primary hyperparathyroidism: diagnosis and management in the older individual. AB - As more and more cases of primary hyperparathyroidism are being detected by screening for serum calcium concentration, the majority of patients are older individuals who are asymptomatic or have symptoms which are difficult to ascribe to hyperparathyroidism. Long-term follow-up has provided evidence that most asymptomatic patients who do not undergo parathyroidectomy will not develop symptomatic complications. Some asymptomatic patients, however, have progression of disease over time. These observations and the lack of reliable predictors of the rate of progression in most patients reinforce the need for careful monitoring in elderly individuals who do not undergo surgery. Biannual measurements of serum calcium concentrations and annual measurements of urinary calcium excretion and bone mineral density should be performed in all patients who are managed conservatively. In elderly patients with symptomatic or complicated primary hyperparathyroidism, parathyroidectomy results in biochemical cure and increased bone density, both at the lumbar spine and the femoral neck, and should be considered. Criteria for surgery include significant hypercalcemia (>1 mg/dl above the upper limit of normal), marked hypercalciuria (>400 mg per day), low bone density, unexplained renal insufficiency and an episode of acute primary hyperparathyroidism. Consideration of parathyroidectomy should also be given to elderly patients with primary hyperparathyroidism who are vitamin D deficient. Radionuclide scanning has become the initial non-invasive study of choice when parathyroid gland localization is necessary before parathyroidectomy; this is generally for fragile patients and reoperative cases. In a subset of older individuals, surgery may not be an option because of coexisting medical problems even though surgical indications are present. PMID- 15362958 TI - Would wider screening for primary aldosteronism give any health benefits? AB - Fifty years ago, Jerome Conn described 'a new clinical syndrome which is designated temporarily as primary aldosteronism' in a young patient with hypertension, severe hypokalaemia and a benign adrenocortical tumour, for whom an adrenalectomy cured both the hypertension and hypokalaemia. His report identified the condition known as aldosterone-producing adenoma, a form of curable hypertension. According to Conn, the prevalence of primary aldosteronism in the hypertensive population referred to his department was 20%, but this estimate was subject to referral bias. Primary aldosteronism has long been considered rare, with an estimated prevalence of 0.5-2% among unselected hypertensive patients. During the past 10 years, however, the apparent prevalence of the condition increased dramatically, up to 30% in some series. Overall, the prevalence of primary aldosteronism in series dealing with at least 100 screened hypertensive patients averaged 6%, with one patient in two harbouring an aldosterone-producing adenoma. This increase in prevalence reflects the fact that hypokalaemic and normokalaemic patients are now screened for primary aldosteronism, with the aldosterone to renin ratio used as a screening tool. The current 'epidemic' of primary aldosteronism raises several questions and concerns. PMID- 15362959 TI - Effects of octreotide on sleep apnoea and tongue volume (magnetic resonance imaging) in patients with acromegaly. AB - OBJECTIVES: Sleep apnoea has been consistently reported to occur in acromegaly. Both obstructive apnoeas, in which apnoeas are due to intermittent obstruction of the upper airways, as well as central apnoeas are known to occur. Because the relationship between disease activity and severity of sleep apnoea is currently unclear, we have performed a prospective study to address this issue. DESIGN AND METHODS: In 14 newly diagnosed patients with active acromegaly (eight females and six males; mean age 57+/-4 years; IGF-I 583+/-48 microg/l; GH 13.5+/-7.0 microg/l (means+/-s.e.m.)), tongue volume and signal intensity of the tongue were examined by magnetic resonance imaging and sleep apnoea was characterised by polysomnography before and after 6 months of treatment with octreotide acetate (Sandostatin LAR 10-30 mg every 4 weeks i.m.). RESULTS: The initial tongue volume was significantly higher in patients with acromegaly (151+/-9 ml; females 133+/ 10 ml; males 172+/-10 ml) in comparison with the body mass index (BMI)- and age matched healthy control group (97+/-5 ml, P<0.001; females 75+/-1 ml, P<0.001; males 120+/-3 ml, P<0.003). After treatment with octreotide, IGF-I was normalised within the age-adjusted normal range in 50% of the patients. In these patients, tongue volume significantly decreased (120+/-14 ml, P<0.05) in comparison with the persistent uncontrolled group of acromegalics (137+/-10 ml, P=not significant). Overall, tongue volume (128+/-8 ml, P<0.05) and the signal intensity ratio of the tongue decreased significantly after treatment with octreotide acetate (120+/-3 vs 105+/-3, P=0.003). The BMI-adjusted tongue volume correlated with IGF-I levels (r=0.60, P<0.002) and the disease duration (r=0.71, P=0.006). At baseline, 50% had obstructive sleep apnoea with a mean respiratory disturbance index (RDI) of >20/h (range 5.1-91.5) and no patient had central sleep apnoea. After 6 months of octreotide treatment, there was a 28+/-10% decrease in RDI. However, RDI did not correlate with IGF-I or GH levels, but correlated positively with BMI (r=0.58, P=0.001) and age (r=0.46, P=0.02). CONCLUSIONS: Obstructive sleep apnoea but not central sleep apnoea frequently occurs in patients with active acromegaly. Successful treatment with octreotide can decrease tongue volume, which may have benefits for coexisting sleep disordered breathing. PMID- 15362960 TI - Efficacy and tolerability of lanreotide Autogel therapy in acromegalic patients previously treated with octreotide LAR. AB - OBJECTIVE: This open label, multicentre study was designed to evaluate the efficacy and tolerability of lanreotide Autogel (L-Autogel) in acromegalic patients over a 24-week period. The outcome of treatment with this new, long acting, aqueous formulation of lanreotide was also compared with the patients' previous treatment with octreotide long acting repeatable (LAR). DESIGN AND METHODS: Twenty-five acromegalic patients (13 males, mean age 51+/-12 years) were switched from octreotide LAR (20-40 mg/4 weeks for at least 6 months) to L Autogel, given deep subcutaneously at a fixed dose of 90 mg/4 weeks. After 12 weeks, the dose of L-Autogel was titrated according to patients' mean GH and IGF I levels at week 8. It was increased to 120 mg/4 weeks if GH>2.5 microg/l or if IGF-I was above the age-adjusted normal range. It was reduced to 60 mg/4 weeks if mean GH<1 microg/l and IGF-I was within the normal range. If the values did not fall within these ranges, the dose remained unchanged at 90 mg. RESULTS: After 24 weeks of treatment with L-Autogel (final doses 60 mg in 3 patients, 90 mg in 4 patients and 120 mg in 18 patients), mean serum GH (2.9+/-2.4 microg/l) and IGF-I concentrations (332+/-193 microg/l) remained statistically unchanged when compared with baseline values under octreotide LAR (GH 2.4+/-1.8 microg/l and IGF I 337+/-201 microg/l, non significant (NS)). There was a significant improvement of the acromegalic symptom score over the study period, from 4.8+/-3.4 to 2.8+/ 2.5 (P<0.001) and a small but significant reduction in the residual pituitary tumour volume (P<0.05). Local side-effects were observed less frequently and no technical problems were encountered with the L-Autogel injections, as opposed to treatment with octreotide LAR (60 difficult injections/150 (P<0.001)). CONCLUSIONS: L-Autogel appears to be as effective as octreotide LAR in lowering GH and IGF-I concentrations in acromegalic patients. This treatment was also well tolerated by the patients, giving fewer local side-effects and technical problems with injections. These advantages may improve the long-term acceptability of medical treatment in acromegaly. PMID- 15362961 TI - Atypical depression in growth hormone deficient adults, and the beneficial effects of growth hormone treatment on depression and quality of life. AB - OBJECTIVE: Some growth hormone deficient adults (GHDAs) have an impaired quality of life, which may improve with growth hormone (GH) treatment. The objective of our study was to make an in-depth psychiatric evaluation of patients with adult onset (AO) and childhood-onset (CO) GH deficiency (GHD), and to assess the time course of changes in their quality of life and symptoms of depression in response to GH treatment. DESIGN: The study design was a 4-month, double-blind, cross over, placebo-controlled trial of GH therapy. METHODS: We used a detailed psychiatric interview to characterise 25 patients with proven GHD at baseline. They were reassessed at monthly intervals during treatment with GH or placebo, using the Nottingham Health Profile and two well-recognised depression rating scales. RESULTS: 11/18 (61%) of the patients with AO-GHD, but 0/7 of the patients with CO-GHD, were found to have atypical depression at baseline. There were significant improvements in the depression rating scale scores after 2 months of GH therapy, with significant improvements in emotional reaction and social isolation scores from 1 month, and in energy levels and sleep disturbance from 2 and 3 months respectively. CONCLUSIONS: The results of our study confirm that a large proportion of GHDAs have unequivocal psychiatric morbidity, and suggest that a response to treatment can be seen after a short trial of GH therapy. We hypothesise that this rapid improvement of symptoms of atypical depression represents a direct central effect of GH therapy. PMID- 15362962 TI - Mutations in the NSD1 gene in patients with Sotos syndrome associate with endocrine and paracrine alterations in the IGF system. AB - OBJECTIVE: To investigate the effect of nuclear receptor Su-var, 3-9, enhancer of zeste, trithorax (SET) domain-containing protein 1 (NSD1) gene alteration in patients with Sotos syndrome on plasma IGFs and IGF-binding proteins (IGFBPs), as well as on the IGF/IGFBP system activity at the tissue level. DESIGN: Twenty-nine patients suspected of Sotos syndrome were divided into two groups: patients with heterozygous deletions or mutations in the NSD1 gene (NSD1(+/-)) (n=11) and subjects without (NSD1(+/+)) (n=18). Plasma samples (n=29) and skin fibroblasts (n=23) were obtained. The results of both groups were compared and related to reference values. METHODS: IGF-I, IGF-II, IGFBP-2, IGFBP-3, IGFBP-4 and IGFBP-6 levels were determined by RIAs. The mitogenic response of fibroblasts to IGFs was investigated by [methyl-(3)H]thymidine incorporation. IGFBP-3 levels in the culture media were measured by RIA. IGFBP-3 mRNA expression was determined by real time RT-PCR. RESULTS: NSD1(+/-) patients showed significantly altered levels of IGF-I (mean-1.2 SDS), IGF-II (-1.2), IGFBP-3 (-1.7), IGFBP-4 (-0.4), IGFBP-2 (+0.8) and IGFBP-6 (+1.5). The NSD1(+/+) patients did not differ from the reference, with the exception of the mean IGFBP-3 level (-1.3). Basal proliferation and mitogenic response to IGFs was diminished in NSD1(+/-) fibroblasts compared with NSD1(+/+) (basal, P=0.02; IGF-I, P<0.001; IGF-II, P=0.02). Compared with control fibroblasts, only the mitogenic response was diminished (basal, P=0.07; IGF-I, P=0.04; IGF-II, P=0.04). A trend of higher IGFBP-3 secretion after IGF-I stimulation (P=0.09) and 3.5-5 times higher mRNA expression of IGFBP-3 in basal conditions was found in NSD1(+/-) fibroblasts in comparison to controls. CONCLUSIONS: NSD1(+/-) patients show endocrine and paracrine changes in the IGF system. These changes may contribute to the abnormal growth pattern. PMID- 15362963 TI - Adult GH deficiency in Japanese patients: effects of GH treatment in a randomised, placebo-controlled trial. AB - OBJECTIVE: To evaluate the influence of factors intrinsic to the Japanese population on consequences of growth hormone deficiency (GHD) and effects of GH treatment in adult Japanese hypopituitary patients with GHD. STUDY DESIGN: A 24 week, randomised, placebo-controlled, double-blind study in 64 patients in Japan, with GHD onset during adulthood (AO; n=27) or childhood (CO; n=37). Body composition measured by dual-energy X-ray absorptiometry (DXA) was evaluated centrally. Serum IGF-I, IGF binding protein-3 (IGFBP-3) and lipid levels were determined centrally. RESULTS: In contrast to Caucasian patients, there were no significant differences before treatment between AO and CO patients for body mass index (BMI), lean body mass (LBM) and fat mass (FM). Baseline BMI was >/=25 kg/m(2) for 32.8% of patients. For all patients combined, a significant increase in LBM and decrease in FM (P<0.001 for each) was seen with GH treatment. Serum IGF-I and IGFBP-3 were significantly lower at baseline in CO compared with AO patients, similar to Caucasian patients, and were increased in both onsets following GH treatment. Serum total and low-density lipoprotein (LDL)-cholesterol concentrations did not differ between AO and CO patients at baseline and were elevated compared with normal ranges. GH-induced decreases were significant compared with placebo for both total (P=0.036) and LDL-cholesterol (P=0.040). Glycosylated haemoglobin was increased with GH compared with placebo treatment (P=0.016) but remained within the upper limit of normal for all patients at endpoint. CONCLUSIONS: Adult Japanese hypopituitary patients with GHD demonstrated obesity relative to healthy Japanese subjects but the clinical presentation differed from that of Caucasian patients. GH treatment improved body composition and serum cholesterol profiles of adult Japanese hypopituitary patients with GHD. PMID- 15362964 TI - Insulin sensitivity in Turner's syndrome: influence of GH treatment. AB - OBJECTIVE: Excessive GH secretion may lead to secondary diabetes mellitus, while prolonged GH treatment may accelerate the onset of type 2 diabetes mellitus in predisposed individuals. Turner's syndrome (TS) patients are a population at risk since they have reduced glucose tolerance (GT) spontaneously and because they are usually treated with high doses of GH. DESIGN AND METHODS: The aim of the study was to evaluate insulin sensitivity (IS) and glucose tolerance (GT) in a group of TS patients treated with GH for a period of 6 years. Forty-seven TS girls were included in the study. GH was administered at a mean weekly dosage of 0.35 mg/kg, injected subcutaneously over 6-7 days. GT was assessed according to the criteria of the Expert Committee on the Diagnosis and Classification of Diabetes Mellitus. IS was evaluated with the quantitative insulin sensitivity check index (QUICK-I). RESULTS: No significant increase of impaired GT was observed in the patients during the follow-up period, while a reduced IS was detected. IS in TS patients was already lower than in prepubertal controls (P<0.001) before starting treatment and further decreased during the first year of therapy (P<0.05), and then remained stable over the following years. No correlation was found between QUICK-I, body mass index, years of treatment, onset and duration of puberty. One patient became diabetic during the course of treatment. CONCLUSIONS: GH treatment in TS girls does not significantly increase the prevalence of impaired GT or type 2 diabetes mellitus, while it does, however, decrease IS. PMID- 15362965 TI - Interest of the androgen deficiency in aging males (ADAM) questionnaire for the identification of hypogonadism in elderly community-dwelling male volunteers. AB - OBJECTIVE: To date, serum free testosterone measurement is considered to be the gold standard for the diagnosis of hypogonadism in elderly males but it is not available to all subjects suspected of a decrease in testicular function. Therefore, we evaluated whether the Androgen Deficiency in Aging Males (ADAM) questionnaire, in its original or in a modified 'quantitative' version (qADAM), could be used as a surrogate to biochemical determinations for the identification of hypogonadism in elderly males. METHODS: 5028 men, aged 50-70 years, spontaneously consulting for the assessment of their gonadal function were studied. ADAM and qADAM, allocating a value of 1 point for any positive answer to each of the 10 questions of the ADAM test, were assessed for their ability to discriminate between males with free testosterone levels below or above 70 ng/l. RESULTS: The sensitivity and specificity of the ADAM score were 81% and 21.6% respectively. The use of ADAM resulted in an appropriate classification of our population in normal or hypogonadal subjects in 44.5% of the cases. The area under the receiver operating characteristics (ROC) curve for the qADAM (0.529) revealed a highly marginal interest of this quantitative approach compared with the original scoring system. CONCLUSIONS: The ADAM test has a high sensitivity to identify aging males with low free testosterone levels. However, due to its lack of specificity, this test cannot be used as a surrogate to serum free testosterone testing for the identification of androgen deficiency in elderly, community-dwelling males. PMID- 15362966 TI - Plasma adiponectin and serum advanced glycated end-products increase and plasma lipid concentrations decrease with increasing duration of type 2 diabetes. AB - OBJECTIVE: To prospectively follow the concentrations of plasma adiponectin (p adiponectin) and serum advanced glycation end-products (s-AGE) in relation to plasma lipids and retinopathy over 3 years in type 2 diabetic patients. DESIGN AND METHODS: P-adiponectin, s-AGE, plasma lipids and diabetic retinopathy were prospectively evaluated in 61 type 2 diabetic patients at baseline and at follow up 3 years later. RESULTS: Mean p-adiponectin (from 8.84+/-5.14 to 11.05+/-6.16 microg/ml; P=0.006) and s-AGE (from 637+/-242 to 781+/-173 ng/ml; P<0.0001) concentrations had increased at follow up. In addition, HbA1c (7.7+/-1.7 to 7.4+/ 1.4%; P=0.0045) and fasting C-peptide (1.00+/-0.38 to 0.81+/-0.35 nM; P=0.019) had decreased and all lipid variables had significantly improved at follow up. P adiponectin correlated inversely with fasting C-peptide (r(s)=-0.273; P=0.045) and low-density lipoprotein (LDL)/high-density lipoprotein (HDL) ratio (r(s)= 0.362; P=0.011), and directly with plasma HDL cholesterol (r(s)=0.381; P=0.005) at follow up. Analysis of variance with adiponectin and s-AGE as dependent variables and fasting C-peptide, plasma HDL and plasma LDL cholesterol as covariates demonstrated that the increase in s-AGE was independent (P=0.001) and the increase in p-adiponectin dependent on covariate changes (P=0.862). There was a slight correlation between s-AGE at baseline versus the degree of retinopathy at follow up (r(s)=0.281; P=0.0499). CONCLUSION: Both p-adiponectin and s-AGE increased during the 3 years. The increase in p-adiponectin was explained by improvements in insulin sensitivity and dyslipidaemia, whereas the increase in s AGE was independent of changes in metabolic covariates. s-AGE increase when the duration of type 2 diabetes increases. PMID- 15362967 TI - PAX8 and peroxisome proliferator-activated receptor gamma 1 gene expression status in benign and malignant thyroid tissues. AB - OBJECTIVE: Genetic alterations involving the thyroid transcription factor PAX8 and the peroxisome proliferator-activated receptor gamma 1 (PPARgamma1) genes have been described in thyroid neoplasms. We investigated in a series of thyroid samples, including 14 normal, 13 hyperfunctioning tissues, 26 follicular adenomas, 21 follicular and 41 papillary carcinomas, both the frequency of the PAX8-PPARgamma1 rearrangement and the expression of the PAX8 and PPARgamma transcripts. METHODS: Using RT-PCR followed by sequencing PCR products, PAX8 PPARgamma1 translocation was not detected in benign tissues nor in papillary carcinomas and was detected in 4 (19%) of 21 follicular carcinomas and in one (4%) of 26 follicular adenomas. RESULTS: Specific real-time quantitative RT-PCR (Q RT-PCR) methods detected high levels of PPARgamma transcripts in follicular carcinomas presenting the rearrangement. Interestingly, the level of PPARgamma transcripts was significantly decreased in papillary carcinomas in comparison with those found in benign adenomas and follicular carcinomas. Finally, PAX8 gene expression was decreased in both papillary and follicular thyroid carcinomas, and in these tumors to the same extent in the presence or absence of the rearrangement. These alterations in both PPARgamma and PAX8 gene expression may explain the poorly differentiated histotype of follicular carcinomas harboring the translocation. Immunohistochemistry showed that nuclear PPARgamma staining was weak in normal tissues, adenomas, papillary carcinomas and in some follicular carcinomas, and strong in the follicular carcinomas positive for the PAX8 PPARgamma1 translocation, but also in some follicular tumors in which no translocation could be evidenced. CONCLUSION: These observations confirm that the PAX8-PPARgamma1 translocation characterizes a subset of thyroid follicular carcinomas but is not a specific marker of carcinoma and that its frequency is lower than that initially reported. Finally, immunohistochemistry is not a reliable method for the specific detection of the translocation, that can be specifically evidenced by Q RT-PCR. PMID- 15362968 TI - Neonatal stimulation of the thyroid gland with iodine or suppression during adolescence with triiodothyronine changes the prevalence of autoimmune thyroiditis in BB rats. AB - OBJECTIVE: Changes in the functional state of beta cells by neonatal stimulation or adolescent suppression have reduced the incidence of type 1 diabetes mellitus in animal models. The aim of this study was to evaluate the effect of manipulation of the activity of the thyroid gland by neonatal stimulation or by adolescent suppression on the prevalence of spontaneous autoimmune thyroiditis (AIT) in rats. METHODS: Bio-Breeding/Worcester (BB) rats were treated neonatally with sodium iodine (NaI) or thyroid stimulating hormone (TSH), or during adolescence by triiodothyronine (T(3)), and the lymphocytic infiltration in the thyroid gland was evaluated. RESULTS: Neonatal treatment with NaI decreased the prevalence of AIT to 32+/-9% compared with 66+/-5% in the controls (P<0.002), mainly caused by a reduction among the female rats (13+/-9% vs 52+/-8%, P<0.006). TSH had no effect. Post neonatal suppression of the thyroid gland by T(3) had a biphasic response. Early in adolescence the overall prevalence was 14+/-7% compared with 66+/-5% in the controls (P<10(-5)); for female rats AIT was prevented (0+/-0%) compared with 52+/-8% in the controls (P<0.0003) and in male rats the values were 29+/-13% compared with 80+/-6% in the controls (P<0.001). Treatment with T(3) later in adolescence increased the overall prevalence to 81+/ 7% compared with 66+/-5% in the controls (not significant). For female rats the prevalence increased to 78+/-9% compared with 52+/-8% in the controls (P=0.04). The degree of thyroiditis among the affected animals was similar in all groups. CONCLUSION: Neonatal stimulation of the thyroid gland by iodine or early adolescent suppression by T(3) reduced the prevalence of AIT whereas T(3) given later increased the prevalence of thyroiditis in rats. Thyroid activity at various ages seems to be of importance for the development of autoimmune thyroiditis. PMID- 15362969 TI - RGS 2 expression is regulated by TSH and inhibits TSH receptor signaling. AB - OBJECTIVE: A new family of guanosine triphosphatase-activating proteins known as regulators of G protein signaling (RGS) has been found to regulate the desensitization of several G protein-coupled ligand-induced processes. The expression of nine RGS mRNAs was found in human thyroid tissue (RGS 2, 3, 5, 6, 9, 10, 12, 14 and 16). At present, little is known as to whether any of the RGS proteins play a role in TSH signaling. DESIGN AND METHODS: To explore the involvement of RGS proteins in the regulation of TSH receptor (TSHR) signal transduction, mRNA expression levels of the RGS proteins were analyzed after TSH stimulation of human thyroid primary cultures by real-time RT-PCR. Furthermore, the effects of RGS 2 expression on TSHR signaling (cAMP-, inositol-3-phosphate accumulation, TSHR cell surface expression) were studied in COS-7 cells. RESULTS: Only RGS 2 mRNA was found to be regulated by TSH in thyroid primary cultures. Co expression of RGS 2 and TSHR in COS-7 cells reduced the TSHR signaling via inositol-3-phosphate but not via cAMP after stimulation with TSH. CONCLUSION: TSH dependent RGS 2 mRNA expression and the suppression of TSH-G(q)alpha signaling by the overexpression of RGS 2 imply that RGS 2 is involved in TSHR-induced G(q) signal transduction. PMID- 15362970 TI - Immunohistochemical detection of GHRH and its receptor splice variant 1 in primary human breast cancers. AB - OBJECTIVE: GHRH is secreted by the hypothalamus and, upon binding to specific GHRH receptors in the pituitary, stimulates growth hormone (GH) production and release from the pituitary. In addition to this neuroendocrine action, accumulated evidence implies additional roles for GHRH in carcinogenesis in non pituitary tissues. In vitro and in vivo studies have shown that splice variant 1 (SV1) of the GHRH receptor, which is widely expressed in non-pituitary tissues and cancers, can mediate the proliferative effects of GHRH. The aim of the present study was to investigate the operation of an autocrine stimulatory loop between GHRH and SV1 in primary breast tumors. DESIGN: Fifty-three primary breast tumors were evaluated for GHRH and SV1 expression. METHODS: Expression of GHRH and SV1 was assessed by immunohistochemistry using anti-GHRH SV95 and anti-SV1 2317/5 polyclonal antibodies. RESULTS: About 40% of the specimens tested express GHRH and/or SV1 (approx. 25% each), while in 35% of these positive specimens co expression of these antigens was detected (P<0.01). Furthermore, a correlation of GHRH, but not SV1, expression was detected in lobular compared with ductal carcinomas. CONCLUSIONS: These results constitute the first demonstration for the expression of GHRH and SV1 in primary breast cancers, and provide evidence for the operation of an autocrine stimulatory loop between GHRH and SV1 in primary cancers. Our findings indicate that GHRH analogs could have diagnostic and therapeutic applications for the management of breast cancer. PMID- 15362971 TI - Transactivation via the human glucocorticoid and mineralocorticoid receptor by therapeutically used steroids in CV-1 cells: a comparison of their glucocorticoid and mineralocorticoid properties. AB - BACKGROUND: Glucocorticoids (GCs) are commonly used for long-term medication in immunosuppressive and anti-inflammatory therapy. However, the data describing gluco- and mineralo-corticoid (MC) properties of widely applied synthetic GCs are often based on diverse clinical observations and on a variety of in vitro tests under various conditions, which makes a quantitative comparison questionable. METHOD: We compared MC and GC properties of different steroids, often used in clinical practice, in the same in vitro test system (luciferase transactivation assay in CV-1 cells transfected with either hMR or hGRalpha expression vectors) complemented by a system to test the steroid binding affinities at the hMR (protein expression in T7-coupled rabbit reticulocyte lysate). RESULTS AND CONCLUSIONS: While the potency of a GC is increased by an 11-hydroxy group, both its potency and its selectivity are increased by the Delta1-dehydro-configuration and a hydrophobic residue in position 16 (16-methylene, 16alpha-methyl or 16beta methyl group). Almost ideal GCs in terms of missing MC effects, as defined by our in vitro assay, are therefore prednylidene, budesonide, beclomethasone and betamethasone.The MC potency of a steroid is increased by a 9alpha- or a 6alpha fluoro substituent. A hydrophilic substituent in position 16 (like 16 hydroxylation in triamcinolone) decreases both MC and GC properties. As no substituent that leads to an isolated reduction of GC activity could be characterized in our experiments, 9alpha-fluorocortisol, the most frequently used steroid for MC substitution, seems to be the best choice of available steroids for this purpose. PMID- 15362972 TI - Gastric inhibitory polypeptide is the major insulinotropic factor in K(ATP) null mice. AB - OBJECTIVE: ATP-sensitive K(+) (K(ATP)) channels in pancreatic beta-cells are crucial in the regulation of glucose-induced insulin secretion. Recently, K(ATP) channel-deficient mice were generated by genetic disruption of Kir6.2, the pore forming component of K(ATP) channels, but the mice still showed a significant insulin response after oral glucose loading in vivo. Gastric inhibitory polypeptide (GIP) is a physiological incretin that stimulates insulin release upon ingestion of nutrients. To determine if GIP is the insulinotropic factor in insulin secretion in K(ATP) channel-deficient mice, we generated double-knockout Kir6.2 and GIP receptor null mice and compared them with Kir6.2 knockout mice. METHODS: Double-knockout mice were generated by intercrossing Kir6.2-knockout mice with GIP receptor-knockout mice. An oral glucose tolerance test, insulin tolerance test and batch incubation study of pancreatic islets were performed on double-knockout mice and Kir6.2-knockout mice. RESULTS: Fasting glucose and insulin levels were similar in both groups. After oral glucose loading, blood glucose levels of double-knockout mice became elevated compared with Kir6.2 knockout mice, especially at 15 min (345+/-10 mg/dl vs 294+/-20 mg/dl, P<0.05) and 30 min (453+/-20 mg/dl vs 381+/-26 mg/dl, P<0.05). The insulin response was almost completely lost in double-knockout mice, although insulin secretion from isolated islets was stimulated by another incretin, glucagon-like peptide-1 in the double-knockout mice. Double-knockout mice and Kir6.2-knockout mice were similarly insulin sensitive as assessed by the insulin tolerance test. CONCLUSION: GIP is the major insulinotropic factor in the secretion of insulin in response to glucose load in K(ATP) channel-deficient mice. PMID- 15362973 TI - Measurement of coronary vasomotor function: getting to the heart of the matter in cardiovascular research. AB - Measurement of endothelial function in patients has emerged as a useful tool for cardiovascular research. Although no gold standard for the measurement of endothelial function exists, the measurement of flow-mediated dilation in the brachial artery, assessed with Doppler ultrasonography, is the most studied method. However, the assumption that endothelial dysfunction detected in brachial arteries is a manifestation of systemic endothelial dysfunction including the coronary circulation may not be entirely valid. Brachial and myocardial circulations differ in terms of the microvascular architecture, the pattern of blood flow and vascular resistance (e.g. shunt vessels occur in the hand but not in the myocardium), their metabolic regulation, type of receptors that contribute to humoral regulation and the pathways that are activated to induce hyperaemia. In this context, measuring coronary vasomotor function may be more useful than brachial artery measures to predict and assess potential myocardial damage related to limited vascular responsiveness. This review aims to provide an overview of the basic concept of coronary flow reserve and its different modalities of measurement, as well as its utility in cardiovascular research. PMID- 15362974 TI - A novel UBA and UBX domain protein that binds polyubiquitin and VCP and is a substrate for SAPKs. AB - A widely expressed protein containing UBA (ubiquitin-associated) and UBX (ubiquitin-like) domains was identified as a substrate of SAPKs (stress-activated protein kinases). Termed SAKS1 (SAPK substrate-1), it was phosphorylated efficiently at Ser200 in vitro by SAPK3/p38gamma, SAPK4/p38delta and JNK (c-Jun N terminal kinase), but weakly by SAPK2a/p38alpha, SAPK2b/p38beta2 or ERK (extracellular-signal-regulated kinase) 2. Ser200, situated immediately N terminal to the UBX domain, became phosphorylated in HEK-293 (human embryonic kidney) cells in response to stressors. Phosphorylation was not prevented by SB 203580 (an inhibitor of SAPK2a/p38alpha and SAPK2b/p38beta2) and/or PD 184352 (which inhibits the activation of ERK1 and ERK2), and was similar in fibroblasts lacking both SAPK3/p38gamma and SAPK4/p38delta or JNK1 and JNK2. SAKS1 bound ubiquitin tetramers and VCP (valosin-containing protein) in vitro via the UBA and UBX domains respectively. The amount of VCP in cell extracts that bound to immobilized GST (glutathione S-transferase)-SAKS1 was enhanced by elevating the level of polyubiquitinated proteins, while SAKS1 and VCP in extracts were coimmunoprecipitated with an antibody raised against S5a, a component of the 19 S proteasomal subunit that binds polyubiquitinated proteins. PNGase (peptide N glycanase) formed a 1:1 complex with VCP and, for this reason, also bound to immobilized GST-SAKS1. We suggest that SAKS1 may be an adaptor that directs VCP to polyubiquitinated proteins, and PNGase to misfolded glycoproteins, facilitating their destruction by the proteasome. PMID- 15362975 TI - Activation of an alpha2A-adrenoceptor-Galphao1 fusion protein dynamically regulates the palmitoylation status of the G protein but not of the receptor. AB - Post-translational thio-acylation of a fusion protein between the alpha2A adrenoceptor and the alpha subunit of the G protein G(o1) is both dynamic and regulated by agonist binding. Incorporation of [3H]palmitate into the fusion protein was reduced substantially in the presence of the agonist adrenaline. This was dependent on the concentration of adrenaline and correlated with occupancy of the ligand binding site. Both the receptor and G-protein elements of the fusion construct incorporated [3H]palmitate but this occurred more rapidly for the G protein element and regulation of acylation by the agonist occurred only for the G protein. The kinetics of de-palmitoylation of the alpha2A-adrenoceptor Galpha(o1) fusion were accelerated markedly by agonist. Again, this reflected modulation of the G protein but not of the receptor. Agonist-induced regulation of the kinetics of thio-acylation of the G protein was abolished, however, in a mutant unable to bind guanosine 5'-[gamma-[35S]thio]triphosphate ([35S]GTP[S]) in response to adrenaline. Despite the dynamic nature of the post-translational acylation and its regulation by agonist, the ability of adrenaline to activate the G protein, monitored by stimulation of the binding of [35S]GTP[S] to such fusion constructs, was unaffected by the palmitoylation potential of either the receptor or G-protein element. PMID- 15362976 TI - Secondary structure, conformational stability and glycosylation of a recombinant Candida rugosa lipase studied by Fourier-transform infrared spectroscopy. AB - The secondary structure of lipase 1 from Candida rugosa, a model system for large monomeric enzymes, has been studied by FTIR (Fourier-transform infrared) spectroscopy in water and 2H2O. The secondary structure content, determined by the analysis of the amide I band absorption through second derivative and curve fitting procedures, is in agreement with that estimated by X-ray data and predicts, in addition, the existence of two classes of alpha-helices. We have also investigated the enzyme stability and aggregation at high temperature by following the protein unfolding. The thermal stability determined by FTIR is in excellent agreement with the temperature dependence of the lipase activity. Furthermore, new insights on the glycosylation of the recombinant protein produced in Pichia pastoris and on its heterogeneity related to different fermentation batches were obtained by the analysis of the IR absorption in the 1200-900 cm(-1) carbohydrate region. A drastic reduction of the intensity of this band was found after enzymic deglycosylation of the protein. To confirm that the FTIR absorption in the 1200-900 cm(-1) region depends on the carbohydrate content and glycoform distribution, we performed an MS analysis of the protein sugar moieties. Glycosidic structures of the high mannose type were found, with mannoses ranging from 8 to 25 residues. PMID- 15362977 TI - The high concentration of Arg213-->Gly extracellular superoxide dismutase (EC SOD) in plasma is caused by a reduction of both heparin and collagen affinities. AB - The C-terminal region of EC-SOD (extracellular superoxide dismutase) mediates the binding to both heparin/heparan sulphate and type I collagen. A mutation (Arg213- >Gly; R213G) within this extracellular matrix-binding region has recently been implicated in the development of heart disease. This relatively common mutation affects the heparin affinity, and the concentration of EC-SOD in the plasma of R213G homozygous individuals is increased 10- to 30-fold. In the present study we confirm, using R213G EC-SOD purified from a homozygous individual, that the heparin affinity is reduced. Significantly, the collagen affinity of the R213G EC SOD variant was similarly affected and both the heparin and collagen affinities were reduced by 12-fold. Structural analysis of synthetic extracellular matrix binding regions suggests that the mutation alters the secondary structure. We conclude that the increased concentration of EC-SOD in the plasma of R213G carriers is caused by a reduction in both heparin and collagen affinities. PMID- 15362978 TI - Interactions of human replication protein A with single-stranded DNA adducts. AB - Human RPA (replication protein A), a single-stranded DNA-binding protein, is required for many cellular pathways including DNA repair, recombination and replication. However, the role of RPA in nucleotide excision repair remains elusive. In the present study, we have systematically examined the binding of RPA to a battery of well-defined ssDNA (single-stranded DNA) substrates using fluorescence spectroscopy. These substrates contain adducts of (6-4) photoproducts, N-acetyl-2-aminofluorene-, 1-aminopyrene-, BPDE (benzo[a]pyrene diol epoxide)- and fluorescein that are different in many aspects such as molecular structure and size, DNA disruption mode (e.g. base stacking or non stacking), as well as chemical properties. Our results showed that RPA has a lower binding affinity for damaged ssDNA than for non-damaged ssDNA and that the affinity of RPA for damaged ssDNA depends on the type of adduct. Interestingly, the bulkier lesions have a greater effect. With a fluorescent base-stacking bulky adduct, (+)-cis-anti-BPDE-dG, we demonstrated that, on binding of RPA, the fluorescence of BPDE-ssDNA was significantly enhanced by up to 8-9-fold. This indicated that the stacking between the BPDE adduct and its neighbouring ssDNA bases had been disrupted and there was a lack of substantial direct contacts between the protein residues and the lesion itself. For RPA interaction with short damaged ssDNA, we propose that, on RPA binding, the modified base of ssDNA is looped out from the surface of the protein, permitting proper contacts of RPA with the remaining unmodified bases. PMID- 15362979 TI - Processive action of cellobiohydrolase Cel7A from Trichoderma reesei is revealed as 'burst' kinetics on fluorescent polymeric model substrates. AB - Reaction conditions for the reducing-end-specific derivatization of cellulose substrates with the fluorogenic compound, anthranilic acid, have been established. Hydrolysis of fluorescence-labelled celluloses by cellobiohydrolase Cel7A from Trichoderma reesei was consistent with the active-site titration kinetics (burst kinetics), which allowed the quantification of the processivity of the enzyme. The processivity values of 88+/-10, 42+/-10 and 34+/-2.0 cellobiose units were found for Cel7A acting on labelled bacterial cellulose, bacterial microcrystalline cellulose and endoglucanase-pretreated bacterial cellulose respectively. The anthranilic acid derivatization also provides an alternative means for estimating the average degree of polymerization of cellulose and, furthermore, allows the quantitative monitoring of the production of reducing end groups on solid cellulose on hydrolysis by cellulases. Hydrolysis of bacterial cellulose by cellulases from T. reesei revealed that, by contrast with endoglucanase Cel5A, neither cellobiohydrolases Cel7A nor Cel6A produced detectable amounts of new reducing end groups on residual cellulose. PMID- 15362980 TI - C-terminal splicing of NTPDase2 provides distinctive catalytic properties, cellular distribution and enzyme regulation. AB - The present study provides functional characterization of alternative splicing of the NTPDase2 (ecto-nucleoside triphosphate diphosphohydrolase-2) involved in the regulation of extracellular nucleotide concentrations in a range of organ systems. A novel NTPDase2beta isoform produced by alternative splicing of the rat NTPDase2 gene provides an extended intracellular C-terminus and distinguishes itself from NTPDase2alpha isoform in gaining several intracellular protein kinase CK2 (casein kinase 2) phosphorylation sites and losing the intracellular protein kinase C motif. The plasmids containing NTPDase2alpha or NTPDase2beta cDNA were used to stably transfect Chinese-hamster ovary-S cells. Imaging studies showed that NTPDase2alpha was predominantly membrane-bound, whereas NTPDase2beta had combined cell surface and intracellular localization. alpha and beta isoforms showed variations in divalent cation dependence and substrate specificity for nucleoside-5'-triphosphates and nucleoside-5'-diphosphates. NTPDase2beta exhibited reduced ATPase activity and no apparent ADPase activity. NTPDase2 isoforms demonstrated similar sensitivity to inhibitors such as suramin and pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid, and differential regulation by protein kinases. NTPDase2beta was up-regulated by intracellular protein kinase CK2 phosphorylation, whereas NTPDase2alpha activity was down regulated by protein kinase C phosphorylation. The results demonstrate that alternative coding of the intracellular C-terminal domain contributes distinctive phenotypic variation with respect to extracellular nucleotide specificity, hydrolysis kinetics, protein kinase-dependent intracellular regulation and protein trafficking. These findings advance the molecular physiology of this enzyme system by characterizing the contribution of the C-terminal domain to many of the enzyme's signature properties. PMID- 15362981 TI - Cosmetic results of a ventrally based advancement flap for closure of total ear canal ablations in 6 cats: 2002-2003. AB - OBJECTIVE: To report the outcome of a modified total ear canal ablation (TECA) technique to maintain normal ear carriage in cats. STUDY DESIGN: Case series. ANIMALS: Six cats with external ear canal disease. METHODS: Medical records were reviewed to identify cats in which a modified TECA using a single pedicle advancement flap was used to maintain normal ear carriage. Signalment, histopathologic diagnosis, complications, and outcome were retrieved. Owners were contacted to document long-term outcome. RESULTS: Modified TECA procedures (7) were performed in 6 cats. Normal ear carriage was present immediately after surgery and was preserved at follow-up interview. Owners were satisfied with cosmetic appearance. Histopathologic diagnosis of excised ear masses included ceruminous gland adenocarcinoma (2 cats), ceruminous gland adenoma (2 cats, 3 ears), and inflammatory polyp (2 cats). CONCLUSIONS: Modification of the TECA skin incision preserved normal ear carriage in all cats. Owners were satisfied with overall outcome and with cosmetic result. Rate and nature of complications were similar to previously published data. CLINICAL RELEVANCE: A simple modification of the TECA skin incision to create a single pedicle advancement flap maintains normal ear carriage in cats. PMID- 15362982 TI - Evaluation of primary critical ischemia time for the deep circumflex iliac cutaneous flap in cats. AB - OBJECTIVE: To evaluate the primary critical ischemia time for the deep circumflex iliac (DCI) cutaneous flap in cats. STUDY DESIGN: In vivo model. ANIMALS: Thirteen young adult female cats. METHODS: An island skin flap was created on the right side of each cat based on the angiosome of the ventral branches of the DCI vessels. The cats were randomly assigned to a flap ischemia time ranging from 1 to 3 hours in 10-minute intervals. Microvascular clamps were used to occlude the artery and vein for the designated time. Flaps were sutured into position after the ischemic period. On day 3, fluorescein dye was administered and the flaps were evaluated under ultraviolet light to assess percent area of perfusion. On days 7 and 14, the percent area of survival was determined for each flap based on cutaneous morphometry. RESULTS: All flaps had 100 percent area of survival throughout the study. On day 3, all flaps fluoresced uniformly compared with the surrounding skin. On days 7 and 14, all flaps were uniformly viable as confirmed by skin color, consistency, bleeding, and hair re-growth. CONCLUSION: The DCI cutaneous flap in cats can withstand up to 3 hours of ischemia with predictable survival. CLINICAL RELEVANCE: In a clinical setting, high success rates can be expected with microvascular transfer of the DCI cutaneous flap in cats when the ischemia time is <3 hours and precise surgical technique is used. PMID- 15362983 TI - Intraoperative extracorporeal irradiation for limb sparing in 13 dogs. AB - OBJECTIVE: To evaluate extracorporeal intraoperative radiation therapy (IORT) as a treatment method for limb and joint sparing in dogs with appendicular sarcomas in sites other than the distal aspect of the radius. STUDY DESIGN: Retrospective study. ANIMALS: Thirteen client-owned dogs. METHODS: The bone tumor database and medical records (1998-2002) were reviewed for dogs with primary appendicular bone tumors treated with IORT limb-sparing surgery and adjuvant chemotherapy. The segment of bone containing the tumor was isolated from adjacent soft tissue and an osteotomy performed distant to the tumor. The bone segment was exteriorized, irradiated (70 Gy single fraction), and then stabilized with internal fixation. Adjuvant chemotherapy was administered. Lameness was graded and local and distant tumor control was determined. Associations between intra- and postoperative variables with complications and Kaplan-Meier survival analysis for median disease-free interval and survival time were calculated. RESULTS: Limb function was good or excellent in 10 dogs (77%). Postoperative complications (9 dogs, 69%) included deep infection, fracture of the irradiated bone, and implant failure. Surgical failure was more likely if a single implant was used to stabilize the osteotomized bone and if deep infection developed postoperatively. In 3 dogs, tumors recurred locally within bone in the radiation field. The disease-free and overall success rates of extracorporeal IORT for limb and joint preservation were 46% and 54%, respectively. CONCLUSIONS: Extracorporeal IORT provides a novel alternative to traditional techniques for preservation of joint and limb function in dogs with primary appendicular sarcomas. A minimum of 2 implants and intramedullary bone cement should be used to stabilize the osteotomized bone to minimize postoperative complications. Extracorporeal IORT should be used with caution in dogs with tumors of the distal tibia because of a high complication rate. Dogs with tumors in areas of good soft-tissue coverage, such as the humerus and femur, may be good candidates for limb and joint-sparing surgery using extracorporeal IORT. CLINICAL RELEVANCE: Extracorporeal IORT is a surgical technique that can be used for limb and joint salvage in dogs with primary appendicular sarcomas in sites usually not amenable to traditional limb-sparing techniques. PMID- 15362984 TI - Intercalary bone grafts for joint and limb preservation in 17 dogs with high grade malignant tumors of the diaphysis. AB - OBJECTIVE: To evaluate postoperative complications, limb function, and tumor control after intercalary resection and reconstruction for preservation of limb and joint function in dogs with high-grade malignant tumors of diaphyseal bone. STUDY DESIGN: Retrospective study. ANIMALS: Seventeen client-owned dogs. METHODS: The bone tumor database and medical records were reviewed (1986-2002) for dogs with diaphyseal tumors treated with intercalary resection and reconstruction with either an allograft or irradiated autograft. Clinical presentation, diagnostic findings, surgical management, and outcome were determined from medical records and telephone interviews with veterinarians and owners. Statistical analyses included chi2 to test associations between intra- and postoperative variables with complications, and Kaplan-Meier survival analysis for disease-free interval, metastasis-free interval (MFI), and median survival time. RESULTS: Intercalary limb-sparing surgery was performed in 17 dogs with diaphyseal tumors: osteosarcoma (OSA) (15), histiocytic sarcoma (1), and solitary metastasis from a pulmonary adenocarcinoma (1). One dog was excluded from further analysis when the spared limb was amputated 4 days postoperatively because of incomplete tumor resection. In 16 dogs, limb function was good to excellent. Complications occurred in 5 dogs (31.3%) and included superficial infection in 2 dogs (12.5%) and implant failure in 4 dogs (25%). All implant failures occurred in the ulna and there was a significant association between implant failure and non-cemented allografts (P=.042). Non-union of 1 or both osteotomies was diagnosed in 10 dogs (83.3%) and, despite lack of clinical signs in all cases, was significantly associated with the use of intracavitary locally released cisplatin (P=.046) and cemented intercalary grafts (P=.046). Local tumor recurrence was diagnosed in 1 dog (6.3%) and metastatic disease in 12 dogs (75.0%), including 10 dogs with OSA. The median MFI was 137 days. The local disease-free and overall limb-salvage rate was 94% and 100%, respectively. Overall median survival time was 393 days and the median survival time for dogs with OSA was 449 days. CONCLUSION: Intercalary limb sparing surgery results in better postoperative limb function with fewer and less severe complications than historical reports of dogs treated with non-intercalary limb-sparing surgery. CLINICAL RELEVANCE: In dogs with diaphyseal tumors, intercalary limb-sparing surgery preserves normal joint function and results in good to excellent limb use with few complications and good local tumor control. PMID- 15362985 TI - Radiographic and clinical changes of the tibial tuberosity after tibial plateau leveling osteotomy. AB - OBJECTIVE: To investigate radiographic changes of the tibial tuberosity after tibial plateau leveling osteotomy (TPLO) surgery and identify clinical findings and risk factors associated with such changes. STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Hundred and eighty-six client-owned dogs (219 stifles) that had TPLO surgery. METHODS: Patient data retrieved included radiographic changes of the tibial tuberosity during re-examination, age, body weight, whether unilateral or single-session bilateral surgery had been performed, location of the anti-rotational pin, approximate tibial tuberosity area, and approximate average tibial tuberosity width. RESULTS: Fracture with resulting caudal displacement of the proximal tibial tuberosity (1.4%; 3 of 219) occurred less frequently than non-displaced tibial tuberosity fractures (7.3%; 16 of 219). Age, weight, average tibial tuberosity width, location of the anti-rotational pin, and single session bilateral surgery were identified as risks factors for non displaced fracture. Weight divided by the square of the average tibial tuberosity width may be a stronger risk factor than either weight or average tibial tuberosity width alone. CONCLUSIONS: Dogs undergoing single session bilateral TPLO surgery are at greater risk for developing non-displaced tibial tuberosity fractures. The non-displaced tibial tuberosity fracture does not appear to adversely affect outcome or lead to tibial tuberosity avulsion. Significant risk factors for fracture of the proximal tibial tuberosity with caudal displacement were not identified. CLINICAL RELEVANCE: Factors including age, weight, tibial tuberosity thickness, and conditions that may enhance strain on the tibial tuberosity, such as single-session bilateral procedures, may increase risk of fracture. PMID- 15362986 TI - Comparison of 4 blood storage methods in a protocol for equine pre-operative autologous donation. AB - OBJECTIVE: To compare viability of equine whole blood stored by 4 different methods, and to establish optimal storage protocols for an equine autologous blood donation program. STUDY DESIGN: In vitro study of stored equine whole blood. Animals- Six healthy adult horses. METHODS: Blood from each horse was collected into 4 different containers: glass bottles containing acid-citrate dextrose solution (ACD), plastic bags containing ACD, citrate-phosphate-dextrose (CPD), and CPD with supplemental adenine (CPDA-1). Blood was stored for 5 weeks and sampled at 2-day intervals. Standard hematologic and biochemical variables were evaluated, and adenosine-5-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) concentrations were measured and normalized to total hemoglobin content. RESULTS: Plasma hemoglobin, % hemolysis, lactate, potassium, ammonia, and lactate dehydrogenase (LDH) increased, whereas glucose concentration and pH decreased in all stored blood over 5 weeks. There was a temporal increase in hemolysis with all storage methods, but the increase was greatest in glass bottles. Lactate and ammonia were highest in CPD and CPDA-1 samples, indicating more active red blood cell (RBC) metabolism. 2,3-DPG concentrations decreased during storage, but were optimally preserved with CPDA-1. ATP concentrations were significantly higher for blood stored in CPDA-1, and were lowest in glass bottles. CONCLUSIONS: Hematologic and biochemical values measured for blood stored in CPDA-1 are suggestive of improved RBC viability compared with other storage methods. With the exception of ATP, results from stored equine blood were similar to those reported for other species. CLINICAL RELEVANCE: Commercial CPDA 1 bags appear to be the optimal storage method for equine whole blood. PMID- 15362987 TI - Hand-assisted laparoscopic ovariohysterectomy in the mare. AB - OBJECTIVE: To develop a minimally invasive, hand-assisted laparoscopic ovariohysterectomy (HALS-OHE) technique in the mare and to evaluate safety and any associated complications. STUDY DESIGN: Experimental study. ANIMALS: Eight, 11-24-year-old mares with anatomically normal urogenital tracts. METHODS: The surgical technique was developed in 2 non-survival mares and subsequently evaluated in 6 survival procedures. Food was withheld for 48 hours, then mares were anesthetized and positioned in dorsal recumbency for laparoscopic surgery. A hand access device (Omniport) was placed followed by 4 laparoscopic portals. Transection of the ovarian pedicles and broad ligaments was achieved using a combination of a laparoscopic stapling instrument (Endo GIA II), an ultrasonically activated instrument (Harmonic Scalpel), and endoscopic clips (Endo Clip II ML). The genital tract was exteriorized through the laparotomy, and the uterus transected and sutured in a conventional pattern. Horses were evaluated through postoperative day 14 when a necropsy was performed. RESULTS: Four mares recuperated well after surgery, 1 mare was euthanatized because of bilateral femur fracture during anesthetic recovery, and another developed severe pleuropneumonia. At necropsy all but 1 abdominal incision was healing routinely. One mare had abscessed along the celiotomy incision and developed visceral adhesions. Uncomplicated healing of transected mesovarial, mesometrial, and uterine remnants was observed. CONCLUSIONS: Ovariohysterectomy in horses can be accomplished using HALS technique. CLINICAL RELEVANCE: HALS-OHE technique represents a minimally invasive and technically feasible alternative for conventional OHE. Careful patient selection and preparation may reduce the complications observed. The HALS technique may be useful in other laparoscopic surgical procedures. PMID- 15362988 TI - An animal model for interface tissue formation in cemented hip replacements. AB - OBJECTIVE: To create a model in sheep for investigation of early changes related to the formation of an interface membrane in hip prosthesis. STUDY DESIGN: Experimental study. ANIMALS: Twenty-four female adult Swiss Alpine sheep. METHODS: Sheep were divided into 2 groups of 12 for unilateral cemented total hip arthroplasty. In Group I, the prosthesis was fixed with retrograde cement gun injection to achieve a complete cement mantle, whereas in Group II a primary cement mantle defect was produced. Groups I and II were further divided into 2 sub-groups with study end points of 2 and 8.5 months after surgery. Radiographs were evaluated postoperatively and at euthanasia for migration of the femoral component and bone resorption. Histologic sections were evaluated semiquantitatively for changes in cell types and numbers, and bone reactions; and quantitatively for size of interface membrane and new bone formation. RESULTS: Radiographically, there tended to be an increase in bone resorption and periosteal bone formation throughout the femoral shaft in Group II compared with Group I, but this was only statistically significant at the region of the femoral neck (R5) at both time periods (P<.05). Semiquantitative histologic evaluation revealed significant increases (P<.05) in cellularity, numbers of fibroblasts, giant cells, macrophages, and mononuclear cells, in Group II primarily at 2 months after surgery. This was also true for interface membrane formation and bone remodeling. Quantitative data showed an increased in the size of the interface membrane and area of bone formation at 8.5 months in Group II. CONCLUSIONS: The cement defect model offered controlled and repeatable production of an interface membrane. The results suggest that a primary cement mantle defect could be a possible trigger for implant instability, eliciting a cascade of biomechanical and molecular events in bone tissue leading to aseptic loosening. CLINICAL RELEVANCE: The results show the effect of defects in the cement mantle in promoting interface membrane formation. Long-term and biochemical studies are required to evaluate the relevance of this interface membrane formation. PMID- 15362989 TI - Transurethral resection in the management of urethral and prostatic neoplasia in 6 dogs. AB - OBJECTIVE: To assess cystoscopic transurethral resection (TUR) for the palliative management of dogs with neoplastic infiltration of the urethra. STUDY DESIGN: Prospective clinical trial. ANIMALS: Six client-owned dogs. METHODS: Cystoscopic examination and electrosurgical TUR were performed in dogs with urination difficulties caused by prostatic or urethral neoplasia. TUR was performed in a retrograde manner in female dogs and antegrade in male dogs via exploratory celiotomy and ventral cystotomy. Cystoscopic examination was used to determine the extent of neoplastic involvement of the urethra. TUR involved piecemeal removal of neoplastic tissue from the urethral lumen using an electrocautery cutting loop. Hemorrhage was controlled with a cystoscopic cauterized roller ball. In 2 male dogs, intraoperative radiation therapy (IORT) was used to treat both prostatic neoplasia and the sublumbar lymph node bed. Surgical technique, complications, adjuvant treatment, and outcome were recorded. RESULTS: TUR was performed in 3 male dogs with prostatic carcinoma and 2 female dogs with urethral transitional cell carcinoma (TCC). In 1 female dog, TUR was attempted but not successful because of cystoscope diameter. Iatrogenic urethral perforation occurred during TUR in 3 dogs. In 2 dogs, prolonged exposure to lavage fluid resulted in clinical and biochemical abnormalities consistent with TUR syndrome. Dysuria resolved in 5 dogs within 10 days of TUR. Treatment-related complications included urinary tract infection and tumor seeding. Local tumor progression and metastasis occurred in all dogs. CONCLUSIONS: TUR (in combination with chemotherapy+/-IORT) resulted in rapid palliation of urination difficulties in male dogs with prostatic carcinoma. In female dogs with urethral TCC, however, electrosurgical TUR cannot be recommended because of a high intra- and postoperative complication rate with no improvement in postoperative management compared with historical reports of tube cystostomy. CLINICAL RELEVANCE: TUR is a novel alternative for the palliation of male dogs with prostatic carcinoma. In female dogs with urethral TCC, electrosurgical TUR does not provide any advantages compared with tube cystostomy. PMID- 15362990 TI - A vaginal mucosal pedicle flap technique for repair of rectovaginal fistula in mares. AB - OBJECTIVE: To describe the use of a vaginal mucosal pedicle flap for repair of rectovaginal fistulae (RVF) in mares. STUDY DESIGN: Clinical case report. ANIMALS: Three mares with RVF. METHODS: After debriding the RVF, a dorsally based U-shaped mucosal and submucosal pedicle flap (2-4 mm thick) was dissected from the vaginal wall, rotated approximately 90 degrees angle to close the ventral aspect of the fistula and sutured in position with 10-13 single sutures that did not penetrate the rectal mucosa. This positioned the vaginal mucosa so that it was continuous with the rectal mucosa. RESULTS: Two RVF healed by 1st intention whereas for the 3rd RVF, 2 additional single sutures were required to repair partial dehiscence after 10 days. At 6 weeks and 6 months all RVF were healed. One mare subsequently foaled without complications. CONCLUSIONS: RVF can be closed by use of a rotation flap of vaginal tissue. CLINICAL RELEVANCE: Tension free closure of RVF can be achieved by rotating a pedicle flap of vaginal mucosal tissue without undue wound tension, and should be considered as an alternative to conventional appositional techniques. PMID- 15362991 TI - A comparison of knot security of commonly used hand-tied laparoscopic slipknots. AB - OBJECTIVE: To compare the peak force required for failure of hand-tied laparoscopic slipknot ligatures. STUDY DESIGN: In vitro mechanical evaluation of suture material/knot combinations. SAMPLE POPULATION: Four hand-tied laparoscopic slipknots were studied. Three suture materials and 2 suture sizes were evaluated. Twenty samples of each group were tested, for a total of 400 samples. METHODS: The 4S modified Roeder, modified Roeder, and Weston and Brooks laparoscopic slipknots using 0 polyglyconate, 0 polyglycolic acid, 1 polyglyconate, and 1 polydioxanone (PDS) were tested. A vertical distracting force (5 mm/s) was applied until failure. Mean peak force to failure was obtained for each suture material, size, and knot combination. RESULTS: Mean (+/-SEM) peak force to failure of hand-tied ligatures ranged from 156.8+/-9.1 N to 4.8+/-.4 N. The 4S modified Roeder had the highest mean peak force to failure. Ligatures tied with polyglyconate and size 1 suture had the highest mean peak force to failure. CONCLUSIONS: The 4S-modified Roeder was significantly and consistently stronger than the other slipknots. Polyglyconate was superior to other suture materials. The 1 polyglyconate had the highest mean peak force to failure followed by 0 polyglyconate that exceeded the 1PDS. CLINICAL RELEVANCE: The 4S-modified Roeder slipknot using 1 polyglyconate has the greatest breaking strength of sutures tested and should be considered when performing laparoscopic ligatures. PMID- 15362992 TI - Evaluation of arthrocentesis site bacterial flora before and after 4 methods of preparation in horses with and without evidence of skin contamination. AB - OBJECTIVE: To evaluate the effectiveness of four methods of povidone-iodine preparation on skin bacterial flora of arthrocentesis sites, in horses, with and without evidence of skin contamination. STUDY DESIGN: Prospective randomized study. ANIMALS: Twenty-four adult horses. METHODS: Horses were assigned to either the clean or contaminated group based on housing environment and visual evidence of contamination. Using a moist sterile swab, microbial culture samples were obtained from the skin over the distal interphalangeal joints immediately before and after preparation. Each site was aseptically prepared with 1 of 4 povidone iodine techniques: 10-minutes scrub, 5-minutes scrub, three 30-second scrubs, or commercial one-step iodophor surgical solution. Colony forming units (CFUs) were determined for each sample, 24 hours after inoculation, on blood agar plates. RESULTS: Mean (+/-SD) pre-scrub CFUs/mL was significantly higher in the contaminated group (9588.33+/-1223.65) compared with the clean group (4489.00+/ 3842.03) (P<.01). After preparation of the arthrocentesis sites, there were no significant differences in post-scrub CFUs/mL among the 10 minutes (mean clean, 46.00+/-64.36; mean contaminated, 28.67+/-18.04), 5 minutes (mean clean, 84.17+/ 109.80; mean contaminated, 40.33+/-44.52), three 30 seconds povidone-iodine scrubs (mean clean, 95.50+/-172.29; mean contaminated, 46.67+/-56.94), or application of a commercial one-step iodophor surgical solution (mean clean, 102.17+/-161.78; mean contaminated 117.67+/-143.78); or between the clean (81.96+/-131.69) and contaminated groups (58.33+/-85.90) (P<.01). CONCLUSIONS: Preparation of the distal interphalangeal joint arthrocentesis site with each of these techniques significantly reduces the bacterial flora to a similar level for arthrocentesis in horses with and without evidence of skin contamination. Clinical Relevance- Aseptic preparation of the skin over the distal interphalangeal joint may be accomplished with any of these techniques. PMID- 15362993 TI - An in vitro study to determine the effectiveness of a patellar ligament/fascia lata graft and new tibial suture anchor points for extracapsular stabilization of the cranial cruciate ligament-deficient stifle in the dog. AB - OBJECTIVE: (1) To determine whether an extracapsular patellar ligament/fascia lata graft would provide stability in the cranial cruciate ligament (CrCL) deficient stifle comparable with that of the intact stifle. (2) To determine if different tibial anchor points would enhance stability of the CrCLdeficient stifle when compared with the standard fabellar-tibial suture (FTS) placement. STUDY DESIGN: Experimental. ANIMALS: Twenty-eight canine cadaver hind limbs. METHODS: Stifles were mounted in a jig and tested between loads of -65 and 80 N. After testing the intact CrCL, 4 stabilization techniques were tested after CrCL transection: lateral graft technique (LGT) and 3 FTS with different tibial anchor points. RESULTS: There were no significant differences in displacement between the LGT and standard FTS, between the LGT and the intact CrCL, or between the FTS and the intact CrCL, in either the Securos or the Screw-washer experiments. Stiffness of the intact CrCL was significantly greater than that of any stabilization technique and the cut CrCL. The standard FTS showed the least displacement of all suture stabilization techniques. Differences in stiffness were not significant between the suture stabilization techniques. CONCLUSIONS: Securely anchored, the LGT results in a reduction in drawer motion similar to that of the intact CrCL and the standard FTS. Altering the tibial anchor point for the FTS does not improve stiffness or enhance stabilization of the CrCL deficient stifle. CLINICAL RELEVANCE: The LGT could be used for the treatment of CrCL ruptures in the dog. A clinical study is recommended. PMID- 15362994 TI - A prospective study of postoperative surgical site infections in dogs and cats. AB - OBJECTIVE: To assess postoperative surgical site infection (SSI) rate and to identify associated predictive factors. STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs and cats that had surgery (1010 interventions) during 58 weeks from April 1999 to June 2000. METHODS: Data sheets were completed by clinicians. Patients were controlled for clinical evidence of SSI at suture removal. Two definitions of SSI ("infection" and "infection/inflammation") were developed specifically for this study and used for statistical analysis. Logistic regression models were built in order to identify significant predictive factors for SSI. RESULTS: Wounds with "infection/inflammation" occurred in 5.8% and "infected" wounds in 3% of patients. The outcome "infection" was associated with 3 major risk factors (duration of surgery, increasing number of persons in the operating room, dirty surgical site) and 1 protective factor (antimicrobial prophylaxis). The outcome "infection/inflammation" was associated with 6 significant factors (duration of anesthesia, duration of postoperative intensive care unit stay, wound drainage, increasing patient weight, dirty surgical site, and antimicrobial prophylaxis). CONCLUSIONS: SSI frequency in companion animals is comparable with the frequency observed in human surgical patients. Several significant predictive factors for SSI in small animals surgery were identified. CLINICAL RELEVANCE: Baseline information for SSI surveillance in our hospital and for comparison with other studies was defined. The factors identified may help to predict infections in surgical patients and to take adequate preventive measures for patients at risk. PMID- 15362995 TI - Effects of ovariohysterectomy on canine blood neutrophil respiratory burst: a chemiluminescence study. AB - OBJECTIVE: To examine blood neutrophil counts and luminol-enhanced chemiluminescence (CL) responses in dogs undergoing ovariohysterectomy (OH), premedicated with 2 different drugs. STUDY DESIGN: Randomized clinical study. ANIMALS: Forty-two healthy client-owned bitches. METHODS: Dogs had OH under isoflurane anesthesia with either acepromazine or medetomidine, both in combination with butorphanol, administered as preanesthetic medication. Blood samples were collected when the dog was admitted, at the end of surgery, and the next day (approximately 20 hours after surgery). Blood neutrophils were counted automatically, and neutrophil oxidative activity was assessed by measuring blood CL responses (induced by opsonized zymosan and enhanced by luminol) at 37 degrees C for 40 minutes. RESULTS: Number of circulating neutrophils was significantly increased the day after surgery reflected by enhanced blood CL responses. Neutrophil CL, however, was not significantly altered. No significant differences were detected for perioperative Polymorphonuclear neutrophil (PMN) characteristics between the 2 preanesthetic regimens. CONCLUSIONS: In conclusion, despite clearly increasing the number of circulating neutrophils, OH did not significantly affect neutrophil respiratory burst, as measured by whole-blood CL responses. CLINICAL RELEVANCE: Surgical operation of moderate intensity (e.g., OH) did not significantly alter one of the important immune functions, neutrophil oxidative activity. Further studies are warranted to confirm the significance of this finding, and to assess the value of following this variable in different animal patient populations. PMID- 15362996 TI - Occult subchondral osseous cyst-like lesions of the equine tarsocrural joint. AB - OBJECTIVE: To report the clinical signs, diagnosis, management, and outcome of horses with occult sub-chondral osseous cyst-like lesions of the tarsocrural joint. DESIGN: Retrospective study. ANIMALS: Twelve horses with subchondral osseous cyst-like lesions (SOCLs) in the tarsocrural joint. METHODS: Information about history, examination findings, diagnostic techniques, and surgical procedures as obtained from medical records. Outcome was determined by patient re evaluation and telephone communication with the referring veterinarian, owner, or trainer. RESULTS: Horses were aged from 3 to 29 years. Lameness (2 weeks to 1 year) varied from moderate to severe. Synovial fluid analyses (9 horses) yielded changes consistent with suppurative inflammation and positive bacterial culture was obtained in 4 horses. Eight of 9 horses that had intra-articular analgesia had a dramatic reduction in lameness. No abnormalities were detected on tarsal radiographs in 10 horses. Scintigraphy identified foci of increased radiopharmaceutical uptake in the distal tibia or talus of all horses, and the lesion was further characterized by computed tomography in 7 horses. SOCLs were identified in the medial malleolus (5), intertrochlear groove of the talus (4), lateral malleolus (2), and distal intermediate ridge of the tibia (1). One horse was euthanatized, 6 horses had surgical debridement, and 5 horses were managed conservatively. Four horses treated surgically (67%) returned to soundness. Conservatively treated horses continued to exhibit lameness but 2 were sufficiently sound for light pleasure riding. CONCLUSIONS: Occult lesions of the tarsus not visible on radiographs can be detected by computed tomography and scintigraphy and may be a source of lameness. CLINICAL RELEVANCE: SOCL, possibly of septic origin, should be a differential diagnosis for persistent lameness localized to the tarsocrural joint without radiographic abnormalities. Surgical debridement of the lesions may offer the best prognosis for a return to athletic soundness. PMID- 15362999 TI - Mesothelial cell proliferation and apoptosis. AB - Mesothelial cells line the pleural and peritoneal surfaces, where under normal conditions they proliferate and undergo cell death at a slow rate, thereby maintaining a constant number of cells. These tightly regulated processes are disrupted in malignancy. By developing a better understanding of the mechanisms that regulate cell proliferation and apoptosis in mesothelial and mesothelioma cells, we may be able to develop more effective therapeutic agents that target specific steps in these pathways to induce apoptosis more efficiently. This paper reviews our current knowledge of the signaling pathways involved in the regulation of mesothelial cell proliferation and apoptosis. The latest advancements in identifying proteins that play key roles in the resistance to apoptosis are highlighted. PMID- 15363000 TI - Radiology in pleural disease: state of the art. AB - Diseases of the pleura and pleural space are common and present a significant contribution to the workload of the chest radiologist. The radiology department plays a crucial role in the imaging and management of pleural disease. This review aims to describe and illustrate the appearances of common pleural pathologies on various radiological modalities including plain film, ultrasound, CT, magnetic resonance imaging and positron emission tomography. The review will also address the state-of-the-art techniques used to image pleural disease and discuss image-guided intervention in the management of pleural disease. PMID- 15363001 TI - Molecular epidemiology of tuberculosis and recent developments in understanding the epidemiology of tuberculosis. AB - The development of molecular typing techniques during the last decade has led to remarkable improvements in epidemiological studies of tuberculosis (TB) and other infectious disease. In this paper, the burden of TB disease, current issues about disease control in developed countries including Australia and the contribution of molecular typing to improved understanding of TB epidemiology are reviewed. PMID- 15363002 TI - Relationship between arterial blood gas values, pulmonary function tests and treadmill exercise testing parameters in patients with COPD. AB - OBJECTIVE: There have been controversial reports regarding the relationship between exercise tolerance and resting pulmonary function in patients with COPD. The aim of this study was to examine the relationship between resting pulmonary function tests (rPFT) and cardiopulmonary exercise testing parameters (CETP) and their value in estimating exercise tolerance of patients. METHODOLOGY: In total, 45 patients with COPD (nine females, 36 males; mean age 61.2 +/- 11.2) and 21 healthy subjects (four females, 17 males; mean age 60.3 +/- 9.7) as a control group were studied. COPD patients (group I) were divided into three subgroups according to their FEV(1) (mild/group II: FEV(1) 60-79% of predicted; moderate/group III: FEV(1) 40-59%; severe/group IV: FEV(1) < 40%). In controls FEV(1) was >/= 80%. RESULTS: There were significant correlations between FEV(1) and CETP in group III (maximal O(2) consumption (mVO(2)), r= 0.35, P < 0.005; total treadmill time (TTT), r= 0.31, P < 0.01; total metabolic equivalent values (TMET), r= 0.29, P < 0.01)) and in group IV (mVO(2), r= 0.49, P < 0.001; TTT, r= 0.45, P < 0.005; TMET, r= 0.31, P < 0.01; peak heart rate (pHR), r= 0.29, P < 0.02; frequency of ventricular extrasystole (fVES), r=-0.27, P < 0.05). Additionally, in group IV there were significant correlations between PaO(2) and CETP (mVO(2), r= 0.41, P < 0.02; TTT, r= 0.38, P < 0.03; TMET, r= 0.31, P < 0.05; pHR, r= 0.29, P < 0.05; fVES, r=-0.28, P < 0.05). CONCLUSION: There are significant correlations of resting FEV(1)% predicted and PaO(2) values with CETP in patients with moderate and severe COPD and these parameters may also have a role as indicators of exercise tolerance in these COPD patients. PMID- 15363003 TI - Determinants of cardiorespiratory fitness in patients with chronic obstructive pulmonary disease, focusing on activities parallel to daily living. AB - OBJECTIVE: The purpose of this study was to examine whether activities parallel to daily living (APDL) constitute a determinant index of cardiorespiratory fitness in patients with chronic obstructive pulmonary disease (COPD). METHODOLOGY: Four functional physical fitness parameters were investigated in 38 male patients (mean age 69.8 +/- 6.7 years) with moderate to severe COPD. The parameters measured were muscular strength (grip strength), muscular endurance strength (arm curl, keeping a half-squat position) and regulation (walking around two cones). In addition, cardiorespiratory fitness was measured using a 6-min walking distance test (6MWD) and peak oxygen uptake (VO(2peak)) was measured during bicycle ergometer testing. Maximal inspiratory pressure (MIP) and maximal expiratory pressure were measured so as to quantify respiratory muscle strength. RESULTS: There were significant correlations (P < 0.05) between the VO(2peak), muscular endurance, pulmonary function and respiratory muscle strength. There were also significant correlations of VO(2peak) to muscular endurance, muscular strength, muscular endurance strength and regulation. In stepwise multiple regression analysis, per cent predicted forced expiratory volume in 1 s and MIP appeared to be significant determinants of VO(2peak), showing a total variance of 56% (P < 0.05). For the 6MWD, the significant determinants were forced vital capacity, MIP and performance in the half-squat test, showing a variance of 59% (P < 0.05). CONCLUSION: The results suggest that cardiorespiratory fitness is strongly affected by MIP, pulmonary function and muscular endurance strength, as APDL depend on lower-limb use. PMID- 15363004 TI - Clinical phenotypes of COPD: results of a Japanese epidemiological survey. AB - OBJECTIVE: The Global Initiative for Obstructive Lung Disease characterizes COPD as airflow limitation caused by parenchymal destruction and/or small airway disease. This report characterizes the clinical features of these two phenotypes of COPD in Japan. METHODOLOGY: COPD was diagnosed by spirometric airflow limitation (FEV(1)/FVC < 70%), and all subjects underwent chest CT scanning. Patients with diffuse low attenuation areas (LAA) on CT scan were categorized as the emphysema-dominant phenotype; those with little LAA were categorized as the airway disease-dominant phenotype. The two groups were compared to identify significant clinical or demographic differences. RESULTS: Of the 1438 patients analysed, 1294 (90%) were classified as having an emphysema-dominant phenotype and 144 (10%) as having an airway disease-dominant phenotype. The airway disease dominant phenotype was: more common than the emphysema-dominant phenotype in women (15%vs. 7%, P < 0.01) and in non-smokers (6%vs. 2%, P < 0.05); was more commonly complicated by asthmatic features (35%vs. 21%, P < 0.01); and had higher IgE and eosinophil levels (P < 0.05) and less lung function impairment. CONCLUSION: This analysis is the first to clinically define two phenotypes of COPD in a Japanese epidemiological survey. There appear to be striking differences as well as overlap between these two groups. Further research is warranted to determine the significance of COPD phenotypes. PMID- 15363005 TI - Improved activities of daily living, psychological state and health-related quality of life for 12 months following lung volume reduction surgery in patients with severe emphysema. AB - OBJECTIVE: The aim of the present study was to evaluate the effect of lung volume reduction surgery (LVRS), with an emphasis on improvement in activities of daily living (ADL), psychological state and health-related quality of life (HRQL), for 12 months in patients with severe emphysema. METHODOLOGY: Eighteen male patients (mean age +/- SD: 65.2 +/- 6.4 years) who underwent LVRS following pulmonary rehabilitation (LVRS group) and 12 patients (67.0 +/- 8.1 years) who were medicated and underwent pulmonary rehabilitation (non-LVRS group) were studied. LVRS was performed by video-assisted thoracoscopic surgery. Serial measurements of lung function, 6-min walking distance (6MWD), ADL, HRQL and psychological state scores were performed before surgery and 3, 6, and 12 months after surgery (in the LVRS group), or on the day of discharge (in the non-LVRS group). RESULTS: As well as an improvement in FEV(1) and 6MWD, ADL scores were significantly improved in the items of 'face washing and teeth brushing', and 'indoor walking' at 3 months after LVRS (P < 0.05). At 12 months, an improvement was still found in 'indoor walking' and 'bathing'. HRQL scores were ameliorated at 3-12 months after LVRS. In the non-LVRS group, ADL and HRQL scores failed to improve in any items during the 12 months of observation. Psychological state scores were maintained in the LVRS group and were better than those in the non-LVRS group at 12 months (P < 0.05). CONCLUSIONS: Lung volume reduction surgery improves not only lung function and exercise performance but also ADL, HRQL and psychological state for at least 12 months. PMID- 15363006 TI - Short- and long-term hospital and community exercise programmes for patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: Pulmonary rehabilitation in patients with COPD has been shown to be beneficial but the optimal setting is not known. In the present study, the efficacy of a short-term community-based exercise programme was compared with a standard hospital outpatient programme. Additionally, the usefulness of community or home programmes in maintaining improvements in the longer term was studied. METHODOLOGY: Forty-three patients with moderate to severe COPD were randomized to one of the following three groups: a 3-month hospital programme then a 9 month home programme (Hospital/Home); a 3-month hospital programme then a 9-month community programme (Hospital/Community); or a 12-month community programme (Community/Community). The initial 3-month programme was analysed by comparing the Hospital group (Hospital/Home plus Hospital/Community) with the Community group (Community/Community). Six-minute walking distance (6MWD), quality of life (Guyatt chronic respiratory disease questionnaire, CRQ) and lung function were measured at 0, 3, 6 and 12 months and results were analysed using the Wilcoxon rank sum test. RESULTS: At 3 months, there was a significant improvement from baseline in 6MWD in the Hospital group (81.3 +/- 18.3 m, P < 0.05, anova) but not the Community group (14.4 +/- 28.5 m, not significant). The difference between the groups was not significant (P = 0.058). At 3 months, there was a significant improvement in quality of life in the Hospital group (CRQ +16.3 +/- 3.1, P < 0.01, anova) and in the Community group (CRQ +10.2 +/- 4.9, P < 0.05, anova) but the difference between the groups was not significant. Following the initial 3 month programme, the dropout rate was high overall (73% by 12 months), and therefore data from the maintenance programme could not be analysed. CONCLUSIONS: A 3-month community-based exercise programme for patients with COPD did not improve 6MWD. The long-term retention rates in the programmes were poor. PMID- 15363007 TI - Does addition of inhaled steroid to combined bronchodilator therapy affect health status in patients with COPD? AB - OBJECTIVE: Withdrawal of corticosteroid is associated with a deterioration of health status in COPD. In this study the aim was to determine whether high dose inhaled corticosteroid improves quality of life in patients with COPD. METHODOLOGY: In total, 38 male patients with moderate COPD were included in the study. Baseline quality of life scores were determined using a Turkish version of the St George's Respiratory Questionnaire (SGRQ). Patients were randomly divided into two groups. Group 1 consisted of 20 patients who received existing bronchodilator therapy plus inhaled corticosteroid (800 micro g budesonide) for 12 weeks, while 18 patients in group 2 received bronchodilator and placebo. The SGRQ was repeated after the treatment period. RESULTS: All patients were male and mean age was 67 +/- 8.2 years. Symptom, activity, impact, and total scores were assessed and a difference of four units with treatment was considered to be clinically significant. Total score and activity score were decreased by six units and eight units, respectively, in the placebo group while symptom and impact scores did not change significantly. Total scores and the three component scores improved significantly in the corticosteroid group compared to the placebo group (Deltatotal score: -22 in corticosteroid group, -6 in placebo group, P < 0.01). CONCLUSION: Inhaled corticosteroid improved quality of life scores in patients with COPD, without significant improvement in airflow obstruction parameters. Since improvement of health status is one of the important aims in COPD treatment, use of inhaled corticosteroids should be considered from this perspective. PMID- 15363008 TI - Trial of nurse-run asthma clinics based in general practice versus usual medical care. AB - OBJECTIVE: The aim of this study was to assess the ability of nurse-run asthma clinics based in general practice compared with usual medical care to produce at least a moderate improvement in the quality of life of adults with asthma. METHODOLOGY: A randomized controlled trial involving 80 asthma clinic and 90 usual medical care asthma participants, aged 18 years and older was conducted in 11 general practices in Adelaide. The main outcome measure was the St George's respiratory questionnaire (SGRQ), from which quality-of-life scores were used to assess therapeutic benefit. Lung function measurements and health services utilization data were also collected. RESULTS: One hundred and fifty-three participants (90%) were reviewed at follow up after 6-9 months. There was little difference between groups in baseline measures or for the 6-month follow-up outcomes, including the mean difference in total SGRQ scores (-0.5, 95% confidence interval (CI) -4.0, 2.9) and the mean difference in percentage predicted FEV(1) (2.3%, 95% CI -0.7, 5.3 pre-bronchodilator; 0.4%, 95% CI -5.1, 5.9 post-bronchodilator). Trends in health services utilization were noted. CONCLUSIONS: Nurse-run asthma clinics based in general practice and usual medical care were similar in their effects on quality of life and lung function in adults. These findings cannot be generalized to hospital outpatients and other clinics that manage more severe asthmatic patients. PMID- 15363009 TI - Association between stress and asthma symptoms: a population-based study. AB - OBJECTIVE: Although it is believed that stress contributes to asthma, there are no data from studies in the general population to support this belief. To determine whether stress influences asthma in the general population, a database from a nationwide survey to evaluate the relationship between stress and asthma symptoms was used in the present study. METHODOLOGY: A database generated by the National Health and Nutrition Examination Survey I (in South Korea, 1998) was used for this cross-sectional study. Stratified random samples of 9263 subjects, aged 20 years or older, were selected from the entire population of the country; 95% of these subjects responded. Among the responders, 5048 subjects aged 20-44 years were included in the study. Subjects were interviewed using a questionnaire. Multivariate analysis was performed using multiple logistic regression. RESULTS: The prevalence of wheeze and waking in the night because of cough or shortness of breath in the preceding 12 months was 12.9 and 13.5%, respectively. Multivariate analysis showed that the odds ratios for asthma symptoms increased according to the severity of stress reported by the subjects. When stress levels were classified into four categories of little, some, much and very much stress, the odds ratios for wheeze in the preceding 12 months were 1.7 (95% confidence interval (CI) 1.1, 2.5), 2.6 (95% CI 1.7, 4.0) and 3.6 (95% CI 2.1, 5.9) for subjects reporting some, much and very much stress, respectively, relative to those reporting little stress. Using the same stress-level classifications, the odds ratios for waking in the night because of cough or shortness of breath in the preceding 12 months were 1.5 (95% CI 1.0, 2.3), 2.4 (95% CI 1.6, 3.6) and 4.0 (95% CI 2.5, 6.4), respectively. CONCLUSION: This cross sectional study shows an association between stress and asthma symptoms in the general population of South Korea. PMID- 15363010 TI - Age of onset and the characteristics of asthma. AB - OBJECTIVE: The aim of this study was to evaluate the characteristics of asthma in patients with different ages of disease onset. METHODOLOGY: Five hundred and four asthmatic patients (303 males and 201 females) were divided into five groups based on age of onset (group I, age of onset < 15 years old; group II, 15-24 years old; group III, 25-44 years old; group IV, 45-64 years old; and group V, > 64 years old). The relationship between age of onset and the characteristics of asthma, including lung function, reversibility, atopic status and the incidence of coexistent allergic rhinitis was assessed. Multivariate analysis, adjusted for the duration of disease, gender, educational level, smoking status, and degree of previous inhaled corticosteroid treatment, was performed. RESULTS: Thirty per cent of the patients developed asthma before they were 14 years old (group I), and the percentages of those developing asthma at other ages of onset were: group II, 12%; group III, 29%; group IV, 21% and group V, 8%. In all, 57.6% of the asthmatic patients had coexistent allergic rhinitis; younger patients tended to have this comorbidity. The pulmonary function of older onset asthma patients was worse than that of early onset patients. The multivariate analysis showed that there was a positive correlation between the reduction of pulmonary function and duration of the disease (P < 0.001), but there was no relationship between pulmonary function and gender (P = 0.502), educational level (P = 0.734), smoking pack-years (P = 0.902), or degree of use of inhaled corticosteroid treatment (P = 0.586). CONCLUSION: Asthma is a heterogeneous disease with a wide variety of presentations. This study provides information about the disease characteristics and their relationship with age of onset. Further study is necessary to determine why these differences exist. PMID- 15363011 TI - Survey of asthma control in Thailand. AB - OBJECTIVE: The aim of the study was to determine whether asthma management in Thailand is succeeding in achieving the levels of control, specified in national and international asthma guidelines. METHODOLOGY: Adults with asthma in Bangkok, Chiang Mai, Songkhla, and Khon Kaen were interviewed, and we have reported on their asthma severity, morbidity, control, perception of asthma, and healthcare use. RESULTS: A total of 466 asthma sufferers were interviewed. The burden of asthma was high, with 14.8% of respondents being hospitalized for their asthma in the past year. One-quarter of those surveyed had lost workdays as a result of their asthma, and most patients felt that their lifestyle was limited. The majority of respondents had intermittent asthma (62.9%), 10.5% had mild persistent asthma, 17.6% had moderate persistent asthma, and 9.0% had severe persistent asthma; increasing severity was significantly associated with increased emergency healthcare use (P < 0.00001). Asthma sufferers greatly underestimated the severity of their condition. Only 36.0% used reliever medication, and use of inhaled corticosteroids was low at 6.7%. Understanding of the inflammatory basis of asthma was poor. Few patients underwent lung function tests or took peak flow meter readings. CONCLUSIONS: The burden of asthma is high in Thailand, and guidelines are not being followed. Encouraging greater use of inhaled corticosteroids will be an important step towards improving asthma control. PMID- 15363012 TI - Adult community-acquired pneumonia in Malaysia: prediction of mortality from severity assessment on admission. AB - OBJECTIVES: Prediction of mortality in patients with community-acquired pneumonia (CAP) can be assessed using clinical severity scores on admission to hospital. The clinical benefit of such tools is untested in Asian countries. The aim of this study was to determine the early adverse prognostic factors in patients hospitalized with CAP in Malaysia and to assess the usefulness of the British Thoracic Society (BTS) severity criteria. METHODOLOGY: A prospective study was undertaken of all adult patients admitted between August 2002 and March 2003 in an urban-based university teaching hospital. RESULTS: One hundred and eight patients (mean +/- SD age 55 +/- 20 years; 58% men) were eligible for the study. Thirteen patients (12%) died in hospital and 95 (88%) survived to hospital discharge. Older age, the presence of chronic illness, severity of comorbidity, reduced oxygen saturation and higher blood urea were associated with mortality during admission. Multivariate logistic regression of these variables identified reduced oxygen saturation as the only independently associated variable. BTS criteria fared poorly in predicting mortality compared with clinical assessment by attending clinicians (36-fold increased risk of death by 'clinical assessment' vs two-threefold by 'BTS criteria'). CONCLUSIONS: In hospitalized patients with CAP, certain factors are adversely associated with mortality during admission. Severity criteria validated in specific countries might not be universally applicable. PMID- 15363013 TI - Normal spirometric reference values for Omani children and adolescents. AB - OBJECTIVES: Normal lung function has been shown to be population specific. The aim of this study was to derive normal reference spirometric values for Omani children and adolescents. METHODOLOGY: Forced vital capacity (FVC), forced expiratory volume in 1 s (FEV(1)), peak expiratory flow and forced mid-expiratory flow were measured in 837 healthy Omani school children aged 6-19 years. Multiple linear regression analysis was performed for each spirometric parameter against age, height and weight for boys and girls separately. RESULTS: All measured spirometric parameters increased with age and height and were significantly higher in boys than girls. Height explained the maximum variance for all parameters. After accounting for height in the prediction equations, the contribution of age and weight was minimal. The expiratory ratio (FEV(1)%FVC) was independent of age and height and its mean values (+/- standard deviation) were slightly higher in girls (91.1 +/- 6.1%) than boys (86.5 +/- 7.1%; P < 0.001). The predicted normal values of the subjects using the derived equations were between 5 and 10% lower than the respective values for subjects in Caucasian sample groups. CONCLUSION: The developed prediction equations can be used in clinical practice in Oman and can be considered for use in neighbouring Arab countries. PMID- 15363014 TI - Computed tomography-guided transthoracic fine-needle aspiration in diagnosis of lung cancer: a comparison of single-pass needle and multiple-pass coaxial needle systems and the value of immediate cytological assessment. AB - OBJECTIVES: The aim of the present study was to compare single-pass needle and multiple-pass coaxial needle systems and to evaluate the value of immediate cytological assessment during the procedure in the diagnosis of lung cancer with CT-guided transthoracic fine-needle aspiration. METHODOLOGY: One hundred and forty-three consecutive patients who underwent CT-guided transthoracic fine needle aspiration were divided into three groups. In the first group (group A, 48 patients), a single-pass needle was used for aspiration, but immediate cytological examination was not performed. In the second group (group B, 48 patients), a single-pass needle was used for aspiration and a pathologist immediately assessed the adequacy of the sample obtained. In the third group (group C, 47 patients), a multiple-pass coaxial needle was used for aspiration and a pathologist immediately assessed the adequacy of the sample obtained. RESULTS: The mean number of fine-needle aspirations was 1.25 in group A, 1.39 in group B and 1.34 in group C (group A vs group B, P = 0.08). The diagnostic accuracy was 83.3, 97.9 and 100.0%, respectively (group A vs group B, P = 0.03; group B vs group C, P > 0.05). Although immediate cytological assessment resulted in adequate samples being obtained from all patients in groups B and C, adequate samples were obtained in 41 of 48 patients (85.4%) in group A (P = 0.004). There was no statistically significant difference among the groups with respect to the rate of pneumothorax. CONCLUSIONS: A single-pass needle technique in transthoracic fine-needle aspiration is preferred because there is no significant difference between single-pass needle and multiple-pass coaxial needle systems with respect to the diagnostic accuracy and the complication rate and, in addition, the single-pass needle has a lower cost. The results of the present study suggest that immediate cytological assessment during the procedure reduces an inadequate sampling rate, thus increasing the diagnostic accuracy of the procedure. PMID- 15363015 TI - Molecular polymerase chain reaction diagnosis of pulmonary mucormycosis caused by Cunninghamella bertholletiae. AB - OBJECTIVE: Mucormycosis is an uncommon but frequently fatal infection caused by strains of mucorales in immunocompromised hosts. In this study, we report a case of pulmonary mucormycosis associated with acute lymphocytic leukaemia caused by Cunninghamella bertholletiae, a rare pathogen. METHODOLOGY: Retrospective analysis of the stored serum, sputum, and necropsy lung tissue samples from this patient, enabled subspecies identification by means of panfungal polymerase chain reaction (PCR), direct DNA sequencing of the PCR products, and homology search with nucleotide basic local alignment search tool. RESULTS: The development of a reliable diagnostic blood test for angio-invasive fungal infections such as mucormycosis is desirable, because the sensitivity of culture for these fungi is extremely low. CONCLUSION: Panfungal PCR on serial serum samples might be useful for the diagnosis of pulmonary mucormycosis. PMID- 15363016 TI - Uncontrolled trial of sparfloxacin in retreatment of pulmonary tuberculosis. AB - OBJECTIVE: The aim of the present report was to study the role of sparfloxacin in retreatment of pulmonary tuberculosis. METHODOLOGY: Sixteen patients with sputum smear-positive pulmonary tuberculosis were given sparfloxacin (200 mg daily), along with isoniazid and para-aminosalicylic acid plus additional drugs where indicated. They were followed up and assessed by sputum-smear microscopy and chest X-ray regularly for the entire duration of follow up, which ranged from 15 to 18 months. RESULTS: Twelve patients completed treatment and achieved a satisfactory response in the form of persistent sputum negativity and radiologically quiescent disease. Two patients defaulted after 6 months and one patient discontinued medications because of severe gastrointestinal intolerance. One patient died before completion of the treatment as a result of the sequelae of the disease. CONCLUSIONS: Sparfloxacin can be used as a companion drug along with other drugs in retreatment of pulmonary tuberculosis. The advantages of this drug are a high safety profile, low cost of treatment and easy availability, which are important determinants, especially in developing countries. PMID- 15363017 TI - Variation in barometric pressure in Melbourne does not significantly affect the BTPS correction factor. AB - The conventional BTPS (body temperature and pressure, saturated with water vapour) correction factor varies with ambient barometric pressure (P(B)) and many lung function laboratories measure P(B) daily. The aim was to investigate whether a fixed value for P(B) could replace daily measurements. P(B) was measured daily over a 12-month period. The highest and lowest values in Melbourne in the last century were also recorded from data published by the Bureau of Meteorology. Using these P(B) values, the BTPS factor was determined for a range of spirometer temperatures and compared to the BTPS factors obtained using a fixed ambient pressure of 101.3 kPa. The mean (SD) P(B) measured over the 12-month period was 102.2 kPa (0.64) with a range of 99.9-103.6 kPa. The level of disagreement between the BTPS factor calculated using a P(B) of 101.3 kPa instead of the measured value was greater at lower temperatures. Over the extremes of P(B) during the last century (98.0-104.3 kPa) the use of a standard pressure (101.3 kPa) produced an error in the BTPS factor of or = 20 mL/h per mmHg) and 12% of the facilities used both high and low flux membranes. The average number of the reuse treatments was 15-fold (range 10-22) and the average of the maximum number of reuse treatments was 20-fold (range 10-50). CONCLUSION: Although dialyser reuse treatments are not a common practice in Korea, the reuse programs are steadily increasing. Strict quality control and further regulations regarding the reuse program should be promptly enacted to provide a better quality of haemodialysis treatment for patients in Korea. PMID- 15363053 TI - Oxidative stress from rapid versus slow intravenous iron replacement in haemodialysis patients. AB - METHODS AND RESULTS: Oxidative stress was examined in 19 erythropoietin-treated haemodialysis patients who were receiving 100 mg of iron sucrose every 2 weeks by two intravenous methods, rapid injection and slow infusion. There were no significant differences in incidence of iron oversaturation state between the two methods. Regarding oxidative stress markers, the values of plasma and red blood cell thiobarbituric acid reactive substances (TBARS) expressed in terms of malonyldialdehyde (MDA) equivalents following the two methods did not increase, and the values of area under the curve (AUC) of both markers were not different between both regimens. Also, there were no significant differences in the values of plasma and AUC of anti-oxidant markers including total anti-oxidant status, reduced thiols, and vitamin E among both periods treated with two intravenous iron methods. CONCLUSION: As such, both intravenous iron methods could be safely used without enhancing oxidative stress in haemodialysis patients. The rapid injection method would be the preferred method of intravenous iron administration because it is more convenient while still retaining the safety profile. PMID- 15363054 TI - What is the practical conversion dose when changing from epoetin alfa to darbepoetin outside of clinical trials? AB - BACKGROUND: The recommended conversion dose for changing from epoetin alfa to darbepoetin alfa is 200 units to 1 microg. However, this may result in the over treatment of uraemic anaemia. METHODS: All in-centre haemodialysis patients (n = 60) were converted from an existing subcutaneous epoetin alfa regimen to weekly intravenous darbepoetin alfa. The protocol for anaemia management included a target haemoglobin (Hb) concentration of 120-130 g/L and a ferritin of 300-600 microg/L. Patient treatments were converted from subcutaneous epoetin alfa to weekly, intravenous darbepoetin alfa at month 0, at a conversion dose of 200 units epoetin alfa to 1 microg darbepoetin. RESULTS: Both Hb and ferritin concentrations remained within the target range, but darbepoetin dosages fell from 50.8 to 42.3 microg/week by month 3 (P = 0.02). The initial conversion factor of 210 units/microg rose to 275 units/microg (P = 0.01) at month 4. No difference in conversion dosage could be determined between patients who were epoetin sensitive (<200 units/kg per week) or resistant (>200 units/kg per week, P = NS). Patients were then switched to fortnightly darbepoetin alfa dosing treatments; the existing weekly dose being doubled and Hb levels fell from 125 to 110 g/L (P < 0.0001), despite an increase in the mean dose from 44.9 to 47.5 microg/week (P = 0.02). CONCLUSION: The original dosage reduction after the switch from epoetin alfa to weekly intravenous darbepoetin alfa may offset the increased relative cost of the latter. When administered weekly and intravenously, darbepoetin alfa maintains Hb at a more favourable conversion rate than is currently recommended. PMID- 15363055 TI - A method for the isolation of glomerular and tubulointerstitial endothelial cells and a comparison of characteristics with the human umbilical vein endothelial cell model. AB - BACKGROUND: Abnormalities in the structure and function of glomerular endothelial cells play a pivotal role in the development of progressive renal disease. The vascular abnormalities observed in the renal tubulointerstitium, however, correlate more strongly with progressive renal failure. Therefore, the successful isolation and culture of human renal microvascular endothelial cells from both the glomerulus and tubulointerstitium are paramount in studying renal disease models. METHODS AND RESULTS: This study describes a simple and reproducible method for the isolation of human tubulointerstitial and glomerular endothelial cells by using immunomagnetic separation with anti-platelet endothelial-cell adhesion (anti-PECAM-1) Dyna beads, followed by manual weeding of mesangial and fibroblast contamination. No significant changes in morphological or immunohistochemical characteristics were observed up to passage two of culture. The in vitro characteristics of the endothelial cells were compared to the renal cortical endothelial cells in vivo and the standard human umbilical vein endothelial cell model (HUVECs). Similar to HUVECs, both populations of renal microvascular endothelial cells had a classical cobblestone appearance, stained positively for von Willebrand Factor and PECAM-1 and negatively for antifibroblast surface antigen and anticytokeratin. Differences in the expression of von Willebrand Factor, Wiebel Palade bodies and Flk-1 staining were observed between glomerular and tubulointerstitial endothelial cells. These immunohistochemical characteristics suggested that tubulointerstital endothelial cells were more closely aligned to HUVECS than to the glomerular endothelial cells. This observation indicated that HUVECs may be a suitable model for determining the tubulointerstitial endothelial response to systemic injury. CONCLUSION: In conclusion, a unique and novel method for the differential isolation of both glomerular and tubulointerstitial endothelial cells has been developed. Significantly, characterization of these populations suggests a role for HUVECS in the study of renal tubulointerstitial disease. PMID- 15363056 TI - Interpreting incidence trends for treated end-stage renal disease: implications for evaluating disease control in Australia. AB - BACKGROUND: Five sources of change modify trends in incidence of treated end stage renal disease (ESRD): (i) demography; (ii) disease control, comprising prevention and treatment of progressive kidney disease; (iii) competing risks, which encompass dying from untreated uraemia or non-renal comorbidity; (iv) lead time bias; and (v) classification bias. Thus, rising crude incidence of treated ESRD may conceal effective disease control when there has been demographic change, lessening competing risks, or the introduction of bias. METHODS: Age specific incidences of treated ESRD in Australia were calculated from Australia and New Zealand Dialysis and Transplant (ANZDATA) Registry data by indigenous/non indigenous status (all causes) and by primary renal disease (non-indigenous only) for two successive decades, 1982-1991 and 1992-2001. RESULTS: We postulate that less competing risks explained much of the increase in treated ESRD in the elderly and Indigenous Australians. The increase in glomerulonephritic ESRD in non-indigenous Australians could be ascribed mainly to immigration from non European countries. There was no significant change in incidence of treated ESRD in Indigenous or non-indigenous persons aged less than 25 years, in non indigenous persons aged 25-64 years for ESRD caused by hereditary polycystic disease or hypertension, or in type 1 diabetics aged over 55 years. End-stage renal disease from analgesic nephropathy had declined. The increase in treated ESRD caused by type 2 diabetic nephropathy appeared to be multifactorial. Lead time/length bias and less competing risks may have concealed a small favourable trend in other primary renal diseases. CONCLUSION: Whether recent disease control measures have had an impact on incidence of treated ESRD is not yet certain, but seems more likely than implied by previous reports. PMID- 15363057 TI - Incidence of end-stage renal disease in overseas-born, compared with Australian born, non-indigenous Australians. AB - BACKGROUND: Barriers to immigration from non-European sources were relaxed in the 1970s. As a result, more Australians are now of Middle Eastern, Asian or Pacific Islander origin, rather than British or European. Currently, overseas-born persons comprise one-third of non-indigenous Australians with end-stage renal disease (ESRD). METHODS: Using data recorded by the Australia and New Zealand Dialysis and Transplant (ANZDATA) Registry, age-standardized incidence rates were calculated for ESRD due to all causes and to certain primary renal diseases for all non-indigenous Australians who were aged over 15 years when first treated for ESRD between 1993 and 2001. Truncated age-standardized incidence rates were calculated for ESRD due to glomerulonephritis by type. RESULTS: Immigrants from the British Isles and 'rest of Europe' had less, and those from the Pacific Island nations, East/South-East Asia, Indian subcontinent, Middle East and Southern Europe more ESRD from all causes than the Australian-born. Two diseases accounted for most of the excess: Type 2 diabetic nephropathy and glomerulonephritis (the latter not significant for the Indian-born). There was a small excess (not always significant) of hypertensive/arteriopathic renal disease in Asian- and Middle Eastern-born persons. The East/South-East Asian-born had the highest rates of ESRD due to mesangial immunoglobulin A (IgA) disease and lupus nephritis, and the Middle Eastern-born the highest rates from focal sclerosing glomerulonephritis. CONCLUSION: For Australians born in the Pacific Island nations, Asia, the Middle East or Southern Europe, excess prevalence of, and/or susceptibility to, diseases that cause ESRD has more than offset any 'healthy migrant' effect. PMID- 15363058 TI - Cochrane systematic reviews of treatments for peripheral nerve disorders. PMID- 15363059 TI - A case of oligosymptomatic botulism. The role of single-fiber EMG in early diagnosis. PMID- 15363060 TI - Sensory CIDP presenting as cryptogenic sensory polyneuropathy. AB - The objective of this study was to report that patients with chronic inflammatory demyelinating polyneuropathy (CIDP) can present with a clinical picture of cryptogenic sensory neuropathy. Patients with distal sensory neuropathy and electrodiagnostic studies that are minimally abnormal or consistent with an axonal pathology are usually diagnosed as having cryptogenic sensory neuropathy if no cause for neuropathy can be found. Some of these patients, however, may have sensory CIDP. We reviewed the records of eight patients with CIDP, diagnosed by sural nerve biopsy, who presented with sensory neuropathy and electrodiagnostic studies that were minimally abnormal or revealed changes consistent with axonal neuropathy. All patients reported distal numbness and paresthesias and, on examination, had predominantly large fiber distal sensory loss and normal muscle strength. In most patients, deep tendon reflexes were reduced or absent. Sural nerve biopsies in all patients were consistent with chronic myelinopathy, with quantitative teased fiber analysis revealing segmental remyelination in 13-40% of the fibers. The four patients who received IVIg therapy had improved sensation and gait. Of the remaining four patients, one is being followed, one had spontaneous remission, one was lost to follow-up, and one, with contraindications to therapy, reported disease progression. Sensory CIDP may present as cryptogenic sensory polyneuropathy with normal or axonal electrophysiologic features. Sural nerve biopsy should be considered in patients with progressive, predominantly large fiber sensory neuropathy of otherwise unknown etiology, as they may have sensory CIDP that responds to therapy. PMID- 15363061 TI - Motor nerve damage is associated with anti-ganglioside antibodies in diabetes. AB - Few reports exist on the association between the humoral immune response to glycolipids and neuropathic findings in diabetes. To address this issue, we assayed serum anti-GM1, GD1b, GD1a, and sulfatides IgG and IgM in a group of 85 non-selected diabetic patients, and correlated these antibodies to clinical and electrophysiological findings. Diabetic patients had higher mean titers of anti GM1 (IgM), GD1b, GD1a, and sulfatide (IgG) antibodies when compared to healthy controls. Patients with a positive titer of anti-ganglioside antibodies had significant alterations of motor conduction parameters. The statistical significance increased when considering those patients with both anti-ganglioside reactivity and a high value for glycosylated hemoglobin. Production of antibodies to ganglioside may follow the exposure of hidden motor nerve epitopes in damaged motor nerves and contribute to the neuropathy. PMID- 15363062 TI - The spatiotemporal characterization of endplate reoccupation, with special reference to the superposition patterns of the presynaptic elements and the postsynaptic receptor regions during muscle reinnervation. AB - In this study, an immunohistochemical investigation was carried out to define spatiotemporal characteristics of superposition patterns of the presynaptic elements and the postsynaptic acetylcholine receptor (AChR) sites during the period of endplate regeneration after sciatic nerve crush. The extent of close correspondence of terminal Schwann cell (TSC)-, or axon terminal-, apposing AChR sites was quantitated with three-dimensional images of neuromuscular junctions (NMJs) taken under confocal laser-scanning microscopy. After 3-weeks post-crush (wpc), reoccupation of regenerating TSCs and later arriving axon terminals proceeded within the scope of previously denervated AChR plaques. During this period, the areas of presynaptic elements and the areas of postsynaptic elements were highly correlated. TSCs rapidly reoccupied a greater part of the postsynaptic receptors. In contrast, there was a slower increase of the contact areas of AChR sites overlapped by the axon terminals. Reoccupation by the presynaptic elements at 20 wpc was almost completed in a majority of NMJs, but some anomalous changes still continued to occur in a small proportion of the NMJs (20-30%). Our results suggest that: (a) with gradual increase of the contact areas between presynaptic and postsynaptic elements, imperfect reinnervation and regeneration, due to spatial mismatching or unbalanced growth between presynaptic and postsynaptic elements, result in sporadic remodeling; (b) the difference in superposition patterns between TSCs and axon terminals depends on the ability of making alignment to the endplate gutters in regenerating NMJs; and (c) a complex set of anatomical relationships among the three endplate components affects the process of endplate reoccupation synthetically. PMID- 15363063 TI - Macrophage appearance in the epineurium and endoneurium of dorsal root ganglion exposed to nucleus pulposus. AB - Recently, it was observed that local application of nucleus pulposus may induce a characteristic reaction at the surface of the dorsal root ganglion (DRG). This change was inflammatory in nature and occurred as early as 1 day after the application of nucleus pulposus. Herniation of the nucleus pulposus was surgically induced in the L4-5 disc in rats. The L4 DRGs were resected 3, 24, and 72 h after surgery and sectioned. The sections were processed for immunohistochemistry using antisera to the macrophage marker ED1 and observed using light microscopy. The appearance of macrophages was confirmed 3, 24, and 72 h after the surgery. Macrophages were mainly distributed in the epineurial space of the DRG 3 h after disc incision and also in the endoneurial tissue 24 and 72 h after disc incision. The immunoreactivity was significantly stronger at 24 and 72 h than at 3 h in the parts of the DRG without apparent changes in the disc incision series (p < 0.01). Within the epineurium adjacent to application of nucleus pulposus, the number was significantly higher at 3 h than at 24 and 72 h (p < 0.05). We conclude that experimental disc herniation with leakage of nucleus pulposus results in macrophage recruitment to the epineurium of the DRG 3 h after disc incision and to the endoneurium 24 and 72 h after disc incision. PMID- 15363064 TI - Neuropathy and cognitive impairment following vaccination with the OspA protein of Borrelia burgdorferi. AB - Neurological syndromes that follow vaccination or infection are often attributed to autoimmune mechanisms. We report six patients who developed neuropathy or cognitive impairment, within several days to 2 months, following vaccination with the OspA antigen of Borrelia burgdorferi. Two of the patients developed cognitive impairment, one chronic inflammatory demyelinating polyneuropathy (CIDP), one multifocal motor neuropathy, one both cognitive impairment and CIDP, and one cognitive impairment and sensory axonal neuropathy. The patients with cognitive impairment had T2 hyperintense white matter lesions on magnetic resonance imaging. The similarity between the neurological sequelae observed in the OspA vaccinated patients and those with chronic Lyme disease suggests a possible role for immune mechanisms in some of the manifestations of chronic Lyme disease that are resistant to antibiotic treatment. PMID- 15363065 TI - Carpal tunnel syndrome in elderly patients: results of surgical decompression. AB - This study prospectively compared preoperative and postoperative (6 months after surgical release) clinical and electrophysiological evaluation and self administered Boston questionnaire (BQ) findings of a series of elderly carpal tunnel syndrome (CTS) patients with those of young and adult patients. Three hundred twenty three consecutive hands (282 patients) underwent surgery between 1997 and 2000. Patients were divided into four age groups: group 1 (20-40 years), 49 hands; group 2 (41-54 years), 96 hands; group 3 (55-69 years), 106 hands; and group 4 (70-90 years), 72 hands. Sex ratio, education, duration and bilaterality of CTS, history of wrist and hand trauma, connective tissue and thyroid diseases, diabetes, polyneuropathy, renal failure, and other nerve entrapment syndromes were recorded. The elderly group (group 4) only had more cases of diabetes than groups 1 and 2 (young and adult patients). Before surgery, elderly patients showed more severe clinical objective and electrophysiological findings than young and adult patients. Preoperative subjective findings (BQ scores) were similar in all four groups. Although their absolute postoperative BQ scores and clinical objective and electrophysiological stages improved, elderly patients showed less improvement in all findings than the 20-54 age groups, presumably due to greater preoperative damage and less repair capacity of the compressed nerve. This, however, is not a contraindication for surgical release in elderly patients. PMID- 15363066 TI - Nerve conduction abnormalities in the trembler-j mouse: a model for Charcot-Marie Tooth disease type 1A? AB - The trembler-j mouse is a spontaneously occurring, demyelinating mutant secondary to a point mutation involving a leucine for proline substitution in the first transmembrane domain of the peripheral-myelin protein-22 (PMP-22) gene. It is considered to be a model for Charcot-Marie-Tooth disease type 1A (CMT1A), largely based upon pathologic observations. However, functional studies demonstrating homology with CMT1A patients have not been documented. Sciatic nerve conduction was performed on 30 and 72-day-old wildtype and trembler-j mice in a blinded fashion. The findings in the mutants in both age groups were consistent with profound demyelination. Trembler-j mice appear to have a greater degree of motor nerve conduction slowing relative to human studies involving patients with PMP-22 gene duplication. Functionally, the trembler-j is a good murine model for CMT1A associated with an identical point mutation but may represent a more severe disease phenotype than CMT1A secondary to PMP-22 gene duplication. PMID- 15363067 TI - Role of insulin-like growth factor-I in the treatment of painful small fiber predominant neuropathy. AB - Idiopathic, painful, small fiber predominant peripheral neuropathy is resistant to symptomatic treatment. Previous treatments have not been directed toward repairing the underlying deficit. Growth factors hold promise as agents to encourage axonal regrowth. In vitro, insulin-like growth factor-I (IGF-I) has been shown to prevent neuronal apoptosis, to increase axonal growth, and to support myelination. Using a double-blind, placebo-controlled design, 40 patients were randomized to treatment with recombinant human IGF-I (0.05 mg/kg twice daily by subcutaneous injection) or placebo for 6 months. There were no significant adverse events and minor adverse events occurred equally in both groups. The primary outcome measure was change in score on an analog pain scale. Secondary endpoints included quantitative sensory testing, quantitative autonomic testing, neuropathy impairment score, nerve conduction studies, and neuropathy symptom and change score. There was no significant difference in the primary endpoint between the two groups. Analysis of secondary endpoints and a global impression of improvement by patients and physicians did not show consistent differences between the groups. IGF-I was safe, but did not improve symptoms in this 6-month trial. PMID- 15363068 TI - A very large Schwannoma originating from the median nerve in carpal tunnel. AB - Schwannomas are common benign nerve tumors occurring in the peripheral nerves. A very large schwannoma of more than 5 years duration, originating from the median nerve in the carpal tunnel in a 38-year-old woman, is reported. There was a painful mass, 60 mm in length and 42 mm in diameter, on the palm without signs of sensory disturbance or atrophy on the thenar muscles. Surgical removal was performed under high-power magnification by separating the sensory and motor fascicles from the tumor. Histological examination resulted in a Schwannoma. At 4 year follow-up, the patient was asymptomatic with excellent relief of symptoms. The tumor did not recur. Although cases have been reported in the literature, this is one of the largest ever described without any neurologic deficit. PMID- 15363069 TI - Influence of gender on arrhythmia characteristics and outcome in the Multicenter UnSustained Tachycardia Trial. AB - INTRODUCTION: Previous studies have demonstrated gender differences in risk of sudden death in patients with ischemic heart disease. The Multicenter UnSustained Tachycardia Trial (MUSTT) evaluated the ability of therapy guided by electrophysiologic (EP) testing to reduce mortality in patients with coronary disease, ejection fraction < or =40%, and spontaneous nonsustained ventricular tachycardia. METHODS AND RESULTS: We analyzed the influence of gender on results of EP testing and outcome of patients enrolled in MUSTT. Women made up 14% of the overall MUSTT population and were less likely than men to have inducible sustained randomizable ventricular arrhythmias (24% vs 36%, P < 0.001). Baseline characteristics differed between men and women. In randomized patients, women were older, more likely to have had an infarction within 6 months, more likely to have a history of heart failure, and more likely to have recent angina prior to enrollment than men (P < 0.05). In the EP-guided therapy group, there was no difference in implantable cardioverter defibrillator implantation rate in men and women (45% vs 53%, P = 0.38). There also were no significant gender influences on risk of arrhythmic death or cardiac arrest (2-year event rate 9% in women and 12% in men, adjusted hazard ratio 0.88) or overall mortality (2-year event rate 32% in women vs 21% in men, adjusted hazard ratio 1.51). CONCLUSION: The outcome and benefit of EP-guided therapy in this trial did not appear to be influenced by gender. However, due to the small numbers of women in the trial, small differences in outcome may not be apparent. Plans for future primary prevention trials should include careful risk stratification of women who less often have inducible sustained ventricular arrhythmias and better left ventricular function despite more frequent heart failure. PMID- 15363070 TI - Prevention of sudden cardiac death: when is a woman just like a man? PMID- 15363071 TI - Differences in tachyarrhythmia detection and implantable cardioverter defibrillator therapy by primary or secondary prevention indication in cardiac resynchronization therapy patients. AB - INTRODUCTION: Although numerous trials have shown benefit of implantable cardioverter defibrillators (ICDs) for either primary or secondary prevention, no trial has prospectively enrolled patients from both indications and analyzed ICD utilization between groups. METHODS AND RESULTS: We performed a retrospective review of MIRACLE ICD, a randomized, prospective double-blind trial of cardiac resynchronization therapy (CRT) in the ICD population. Both secondary prevention (N = 563) and primary prevention patients (N = 415) were enrolled. Subgroup analysis for frequency of ventricular tachycardia (VT) and ventricular fibrillation (VF) episodes and detection accuracy revealed that primary prevention patients had a significantly lower frequency of appropriate episodes (0.09 vs 0.43 episodes/month) at significantly faster cycle lengths (303 +/- 54 ms vs 366 +/- 71 ms, P < 0.0001). These episodes were more likely to be classified as VF by the device and thus receive shock therapy (42% by device classification vs 19% in secondary prevention, P < 0.0001). The absolute rate of inappropriate detections in the primary prevention group per month of follow-up was lower but constituted a much higher proportion of all episodes (30% vs 14%, P < 0.0001). Most inappropriate detections in the secondary prevention group were due to rapidly conducted atrial fibrillation; most in the primary prevention patients were due to sinus tachycardia. CONCLUSION: Patients receiving an ICD for CRT therapy with primary prevention indications have a different clinical arrhythmia course than patients with a history of spontaneous VT/VF. This has implications for the optimal programming of ICDs. Longer-term, prospective evaluation of these differences is warranted and should be investigated in the broader ICD patient population. PMID- 15363072 TI - Myocardial bundles with slow conduction properties are present on the left interventricular septal surface of normal human hearts. AB - INTRODUCTION: The aim of this study was to examine the electrophysiologic characteristics of the normal left interventricular septum (LIVS). METHODS AND RESULTS: We explored the LIVS in search of endocardial potentials following ventricular electrograms during sinus rhythm in 28 patients without structural heart disease. In all patients, low-frequency (mean amplitude: 0.81 +/- 0.33 mV) systolic potentials (LA1) were detected in a basal-to-apical activation sequence (earliest to latest QRS-LA1 interval: 93.3 +/- 10.8 ms to 127.1 +/- 16.6 ms). Programmed stimulation demonstrated decremental conduction properties, and the effective refractory period of the tissue between the ventricular electrogram and LA1 at baseline (376.7 +/- 48.8 ms) was significantly prolonged by disopyramide (421.3 +/- 54.1 ms, P < 0.05 vs baseline). In 21 patients (75%), diastolic potentials (LA2) morphologically similar to LA1 (mean amplitude: 0.52 +/- 0.17 mV) were detected in an apical-to-basal activation sequence (earliest to latest QRS-LA2 interval: 368.9 +/- 32.4 ms to 440.7 +/- 45.8 ms). Para-Hisian pacing with capture of the His bundle showed shorter S-LA1 and S-LA2 intervals compared to the beats without His-bundle capture but with ventricular capture (156.3 +/- 11.2 ms vs 183.2 +/- 12.3 ms, and 385.7 +/- 21.6 ms vs 397.4 +/- 23.4 ms, respectively, P < 0.0001). At the same rate, the LA1-LA2 interval was significantly shorter during right ventricular apical than atrial overdrive pacing (220.4 +/- 23.1 ms vs 261.4 +/- 30.7 ms, P < 0.0001). CONCLUSION: Unique myocardial bundles with slow conduction properties and various electrical connections with the specialized conduction system may exist in the LIVS of normal human hearts. PMID- 15363073 TI - Mysterious potentials in the normal human ventricles: where did they come from? PMID- 15363074 TI - Surface atrial frequency analysis in patients with atrial fibrillation: a tool for evaluating the effects of intervention. AB - INTRODUCTION: The aims of this study were to evaluate (1) principal component analysis as a technique for extracting the atrial signal waveform from the standard 12-lead ECG and (2) its ability to distinguish changes in atrial fibrillation (AF) frequency parameters over time and in response to pharmacologic manipulation using drugs with different effects on atrial electrophysiology. METHODS AND RESULTS: Twenty patients with persistent AF were studied. Continuous 12-lead Holter ECGs were recorded for 60 minutes, first, in the drug-free state. Mean and variability of atrial waveform frequency were measured using an automated computer technique. This extracted the atrial signal by principal component analysis and identified the main frequency component using Fourier analysis. Patients were then allotted sequentially to receive 1 of 4 drugs intravenously (amiodarone, flecainide, sotalol, or metoprolol), and changes induced in mean and variability of atrial waveform frequency measured. Mean and variability of atrial waveform frequency did not differ within patients between the two 30-minute sections of the drug-free state. As hypothesized, significant changes in mean and variability of atrial waveform frequency were detected after manipulation with amiodarone (mean: 5.77 vs 4.86 Hz; variability: 0.55 vs 0.31 Hz), flecainide (mean: 5.33 vs 4.72 Hz; variability: 0.71 vs 0.31 Hz), and sotalol (mean: 5.94 vs 4.90 Hz; variability: 0.73 vs 0.40 Hz) but not with metoprolol (mean: 5.41 vs 5.17 Hz; variability: 0.81 vs 0.82 Hz). CONCLUSION: A technique for continuously analyzing atrial frequency characteristics of AF from the surface ECG has been developed and validated. PMID- 15363075 TI - Clustering of RR intervals predicts effective electrical cardioversion for atrial fibrillation. AB - INTRODUCTION: Atrial fibrillation (AF) is characterized by an irregularly irregular ("random") heart beat. However, controversy exists whether the ventricular rhythm in AF is truly random. We investigated randomness by constructing three-dimensional RR interval plots (3D plots), allowing identification of "clustering" of RR intervals. It was hypothesized that electrical cardioversion (ECV) would be more effective in AF patients with clustering, because clustering might reflect a higher degree of organization of atrial fibrillatory activity. METHODS AND RESULTS: The study group consisted of 66 patients (44 men and 22 women; mean age 68 +/- 11 years), who were referred for ECV because of persistent AF. Twenty-four-hour Holter recordings were used to construct 3D plots by plotting each RR interval (x axis) against the previous RR interval (y axis) and the number of occurrences of each of these x,y combinations (z axis). A clustering index was calculated as the percentage of beats within the peaks in the 3D plot. Based on the 3D plots, clustering of RR intervals was present in 31 (47%) of the 66 patients. ECV was effective in restoring sinus rhythm in 29 (94%) of these 31 patients, whereas sinus rhythm was restored in only 25 (71%) of the remaining 35 patients without clustering (P = 0.020). The clustering index ranged from <2% in the 12 patients with failed ECV to >8% in the 32 patients with sinus rhythm at the end of the study (4 weeks after the ECV); the clustering index in the 22 patients with a relapse of AF after effective ECV was intermediate (P = 0.034 and P = 0.042, respectively). CONCLUSION: This study indicates that ECV is more effective in restoring sinus rhythm in AF patients with clustering compared to patients in whom no clustering is apparent on 3D plots. In addition, the degree of clustering appears to be predictive of the overall outcome of ECV; the higher the degree of clustering, the higher the likelihood of sinus rhythm at follow-up. PMID- 15363076 TI - Automatic recognition of abnormal respiratory events during sleep by a pacemaker transthoracic impedance sensor. AB - INTRODUCTION: A number of pacemakers use transthoracic impedance to derive minute ventilation as a sensor for rate adaptation. Transthoracic impedance is also able to track fluctuations in tidal volume occurring in sleep apnea/hypopnea syndromes (SAS). We evaluated the feasibility of a transthoracic impedance-derived pacemaker algorithm for monitoring sleep respiratory disturbances. METHODS AND RESULTS: Forty-two patients who presented with a conventional indication for DDD pacing or cardiac resynchronization underwent conventional polysomnography 1 month after implantation of a Talent trade mark 3 pacemaker (ELA Medical). The respiratory disturbance index (RDI) stored in the pacemaker memory was compared to the apnea/hypopnea index (AHI) derived from polysomnography. The ability of the pacemaker to identify severe SAS patients (AHI > or = 30) was assessed. A minimal systematic error was observed from a Bland and Altman plot (bias = 0.9 events/hour). The ability of the pacemaker RDI to identify severe SAS patients was determined by analysis of the receiver operator characteristic. A cutoff RDI value of 30.6/hour of recording was found to yield 75% sensitivity, 94% specificity, 75% positive predictive value, and 94% negative predictive value. CONCLUSION: The RDI monitoring function appears to be of value in screening pacemaker patients for SAS. Its performance is comparable to existing simple screening techniques. The ability to permanently monitor respiration, in combination with other diagnostic capabilities (such as Holter functions), presents a unique opportunity to monitor the association between arrhythmias and disturbances of breathing during sleep. PMID- 15363077 TI - "Dormant" pulmonary vein conduction revealed by adenosine after ostial radiofrequency catheter ablation. AB - INTRODUCTION: The endogenous nucleoside adenosine is an important intermediate in cellular metabolism, a regulator of function in many organ systems, and a pharmacologic agent with potent electrophysiologic effects. We studied the effects of adenosine on the activation of the pulmonary veins (PVs) after successful ostial isolation in patients with atrial fibrillation (AF). METHODS AND RESULTS: Twenty-nine patients (21 male; mean age 54 +/- 10 years) with refractory highly symptomatic persistent (n = 9) or paroxysmal (n = 20) AF were included in the study. PV isolation was performed using radiofrequency catheter ablation guided by a multipolar basket catheter (Constellation, Boston Scientific). After successful PV isolation, we studied the effects of intravenous adenosine (12-18 mg) on activation of the upper PVs. A total of 83 PVs were successfully isolated. After adenosine, PV activity was recorded in 10 (34%) of 29 left upper PVs studied and in 3 (13%) of 24 right upper PVs, coupled to atrial activity for 20 +/- 7 seconds (adenosine positive). In 8 (62%) of 13 cases, PV potentials were recorded in the distal electrodes of the basket catheter only. Dissociated PV rhythms (N = 8) present after PV isolation disappeared after adenosine for 18 +/- 7 seconds, even if reconduction was missing. In 14 patients (48%), a second EP study was performed for recurrence of AF. Adenosine-positive PVs had a nonsignificant higher rate of recovery of conduction than adenosine negative veins (71% vs 35%, P = 0.095). CONCLUSION: Adenosine induces transient conduction in 25% of PVs following successful isolation. Further studies are necessary to determine the physiologic or pathophysiologic role of adenosine induced reconduction in human hearts or other organ systems. PMID- 15363078 TI - The puzzle of pulmonary vein electrophysiology: the role of adenosine. PMID- 15363079 TI - Efficacy of repeat pulmonary vein isolation procedures in patients with recurrent atrial fibrillation. AB - INTRODUCTION: Pulmonary vein (PV) isolation is effective in the treatment of most patients with atrial fibrillation (AF). Some advocate the addition of linear ablation techniques to improve efficacy; however, previous studies suggest recurrent PV conduction is responsible for AF recurrence. The aim of this study was to determine the effectiveness of repeat PV isolation in patients with recurrent AF after an initial ablation procedure and to determine if any patient characteristics predict failure of repeat PV isolation procedures. METHODS AND RESULTS: Seventy-four patients with two or more AF ablation procedures using selective PV isolation were included. PV isolation was guided with multielectrode ring catheter recordings, electroanatomic mapping, and intracardiac electrocardiography. Radiofrequency energy was delivered using a 4-mm-tip catheter (maximum 40 W, 52 degrees C); cooled-tip ablation was performed in 10 patients. Linear ablation was not performed. Antiarrhythmic drugs were continued for at least 6 weeks after ablation; AF episodes during this period were censored. Reconnection of one or more segments of previously ablated PVs was observed in 97% of patients; reconnected PVs served as the trigger for AF in 77%. Repeat PV isolation resulted in AF control (cure or 90% reduction in AF episodes) in 64 patients (86%) over a follow-up period of 9.1 +/- 6.7 months. "High-risk" characteristics such as left atrial enlargement, persistent AF, or mitral regurgitation did not predict failure of repeat PV isolation procedures. CONCLUSION: Recurrent AF following selective PV isolation is overwhelmingly associated with PV electrical reconnection. Repeat PV isolation without linear ablation provides effective treatment for recurrent AF in patients in whom an initial PV isolation procedure failed, independent of clinical characteristics. PMID- 15363080 TI - Repeat ablation for atrial fibrillation: just a second go-around? PMID- 15363081 TI - Comparison of efficacy of reverse remodeling and clinical improvement for relatively narrow and wide QRS complexes after cardiac resynchronization therapy for heart failure. AB - INTRODUCTION: Cardiac resynchronization therapy (CRT) has been shown to reverse left ventricular (LV) remodeling and improve symptoms in heart failure patients with wide QRS complexes; however, its role in patients with mildly prolonged QRS complexes is unclear. This study investigated if CRT benefited patients with mildly prolonged QRS complexes >120 to 150 ms and explored if the severity of systolic asynchrony determined such a response. METHODS AND RESULTS: Fifty-eight patients (age 66 +/- 11 years, 66% male) who had undergone CRT were studied prospectively. Of these patients, 27 had QRS duration between 120 and 150 ms (group A), and 31 had QRS duration >150 ms (group B). Tissue Doppler echocardiography and clinical assessment were performed at baseline and 3 months after CRT. Both groups had significant reduction of LV volume and increased ejection fraction, +dP/dt, and sphericity index (all P < 0.05). These improvements were greater in group B and were explained by the higher prevalence of systolic intraventricular asynchrony. Significant reverse remodeling (reduction of LV end-systolic volume >15%) was evident in 46% of group A patients and 68% of group B patients. Improvement in clinical endpoints was observed in both groups (all P < 0.01), although the changes in metabolic equivalent and New York Heart Association functional class were greater in group B. In both groups, systolic asynchrony index (TS-SD) was the most important predictor of reverse remodeling (r =-0.78, P < 0.001) and was the only independent predictor in the multivariate model (beta=-1.80, confidence interval =-2.18 to -1.42, P < 0.001); QRS duration was not. A predefined TS-SD value >32.6 ms had a sensitivity of 94% and specificity of 83% to predict reverse remodeling. Improvement of intraventricular asynchrony after CRT was evident only in responders (P = 0.01). CONCLUSION: Improvement of LV remodeling and clinical status is evident after CRT in heart failure patients with QRS duration >120 to 150 ms. These responders are closely predicted by the severity of prepacing intraventricular asynchrony but not QRS duration. PMID- 15363082 TI - Comparison of conventional and biventricular antitachycardia pacing in a geometrically realistic model of the rabbit ventricle. AB - INTRODUCTION: ICDs often are programmed with antitachycardia pacing (ATP) as the first response to ventricular tachycardia (VT). Many ICDs have an additional lead available for ventricular pacing. We hypothesized that using the additional lead for ATP would improve therapy by advancing the orthodromic wavefront, thereby reducing the size of the excitable gap and inducing block of all reentrant activity. METHODS AND RESULTS: Monomorphic VT was initiated in a thin-walled model of rabbit ventricular myocardium that included an apical infarct and anatomically realistic dimensions. ATP with up to eight pulses was delivered at 90% of VT cycle length to one (conventional) or two (biventricular) stimulation areas. Stimulation areas were adjusted from 0.017 cm2 to 0.169 cm2 to modulate interactions between the antidromic and VT wavefronts, and between the orthodromic wavefront and the VT's refractory region. During conventional ATP, we found that larger stimulation areas terminated the VT in three pulses. Continued pacing after termination caused VT reinitiation in the reversed direction in some instances. With smaller stimulation areas, conventional ATP simply reset the circuit. During biventricular ATP, larger stimulation areas terminated VT in one pulse. There were no instances of reinitiation with reversal. However, with smaller stimulation areas, prolongation of refractoriness near the additional stimulation area facilitated induction of functional reentry with pathways modified by continued pacing. CONCLUSION: Our modeling suggests that biventricular ATP is superior to conventional ATP under conditions where the additional ventricular lead effectively advances the orthodromic wavefront. Failure to achieve this advancement poses a risk of VT acceleration. PMID- 15363083 TI - Localizing and quantifying ablation lesions in the left ventricle by myocardial contrast echocardiography. AB - INTRODUCTION: The inability to determine the extent and intramural depth of ablation lesions can hamper the success of catheter ablation. The study tested the feasibility of differentiating radiofrequency ablation lesions from normal myocardium and quantifying their dimensions by myocardial contrast echocardiography (MCE). METHODS AND RESULTS: In 11 normal dogs, we created 14 focal and 4 linear lesions at different left ventricular sites. MCE was performed both before and after ablation by using an intracardiac echocardiography catheter (9 MHz) and infusing contrast microbubbles through the left coronary artery. We initially used two-dimensional MCE to image focal lesions and subsequently three dimensional MCE to image linear lesions. An independent observer examined the lesion pathology. We found that intracardiac echocardiography alone could not delineate lesion dimensions. However, after ablation, MCE localized the lesions as well-defined, low-contrast areas within the normally opacified myocardium. Lesion dimensions by MCE immediately after ablation and 30 minutes later were similar. In 12 focal lesions, the average maximum depth (5.55 +/- 1.38 mm) and average maximum diameter (10.38 +/- 2.09 mm) by MCE were in excellent agreement with the pathologic depth (5.20 +/- 1.45 mm) and diameter (10.61 +/- 1.67 mm). Two focal lesions could not be detected by MCE and later were found to be superficial. Three-dimensional MCE correctly reconstructed the extent and shape of linear lesions compared to pathology (length: 18.7 +/- 5.7 vs 18.5 +/- 5.6 mm; maximum longitudinal cross-sectional area: 81.2 +/- 9.6 vs 76.0 +/- 10.3 mm2). CONCLUSION: MCE accurately localized and quantified radiofrequency ablation lesions in the normal left ventricle. This new application of MCE may advance ablation for managing ventricular arrhythmias that involve intramural or epicardial regions by providing instantaneous anatomic feedback on the effects of ablation during catheterization. PMID- 15363084 TI - Implementation of novel imaging strategies in complex ablation procedures: will "real-time histology" be destiny? PMID- 15363085 TI - Electroanatomic mapping of the right ventricle in a patient with a giant epsilon wave, ventricular tachycardia, and cardiac sarcoidosis. AB - A 47-year-old man presented with sustained monomorphic ventricular tachycardia of right ventricular origin. Surface ECG recorded during sinus rhythm showed a bizarre "double QRS" pattern. Biventricular cardiomyopathy was found with predominant right ventricular involvement, due to cardiac sarcoidosis. Electroanatomic mapping was used to characterize the right ventricular substrate abnormalities and to decipher the specific activation abnormalities responsible for the ECG findings. PMID- 15363086 TI - Implantable cardioverter-defibrillator malfunction due to mechanical failure of the header connection. AB - Four cases of ICD system malfunction occurring within the Medtronic Marquis family over a limited time frame are reported. These cases shared a common element of defective connections between the header post and the right ventricular pace/sense terminal pin that required intervention and reconnection. Inappropriate shocks occurred in two cases. The circumstances of these cases provide important information about the potential difficulties with these devices and highlight the deficiencies of the current system in tracking and analyzing device-related problems. PMID- 15363087 TI - Orthodromic pacemaker-mediated tachycardia in a biventricular system without an atrial electrode. AB - Pacemaker-mediated tachycardia is a well-known complication of dual-chamber devices. In this report, we describe for the first time a case of orthodromic pacemaker-mediated tachycardia in a patient in whom a biventricular system without an atrial electrode had been implanted. Retrograde atrial activation was directly produced by the dislodged coronary vein electrode in the AV groove, resulting in simultaneous capture of the left atrium and left ventricle. During tachycardia, AV nodal conduction was via the anterograde pathway of the circuit and limited the ventricular response. Subsequently, right ventricular activation was sensed by the right ventricular electrode that triggered biventricular pacing and left atrial capture, perpetuating the tachycardia. Because the left atrial threshold was higher than the left ventricular threshold, the problem could be resolved easily by lowering the output of the coronary vein electrode. PMID- 15363088 TI - Variation of HA intervals in atrioventricular nodal reentrant tachycardia with atrioventricular block: what is the mechanism? PMID- 15363089 TI - Mechanism of transition from wide to narrow orthodromic atrioventricular reciprocating tachycardia. PMID- 15363090 TI - Role of imaging techniques in preparation for catheter ablation of atrial fibrillation. PMID- 15363091 TI - Three-dimensional and endocardial imaging of the coronary sinus for cardiac resynchronization therapy. PMID- 15363092 TI - Ventricular wall motion using three-dimensional imaging. PMID- 15363093 TI - 10th Biennial International Gynecologic Cancer Society Meeting (IGCS). Edinburgh, Scotland. October 3-7, 2004. Abstracts. PMID- 15363094 TI - Cytotoxic effects of gemcitabine-loaded liposomes in human anaplastic thyroid carcinoma cells. AB - BACKGROUND: Identification of effective systemic antineoplastic drugs against anaplastic thyroid carcinomas has particularly important implications. In fact, the efficacy of the chemotherapeutic agents presently used in these tumours, is strongly limited by their low therapeutic index. METHODS: In this study gemcitabine was entrapped within a pegylated liposomal delivery system to improve the drug antitumoral activity, thus exploiting the possibility to reduce doses to be administered in cancer therapy. The cytotoxic effects of free or liposome entrapped gemcitabine was evaluated against a human thyroid tumour cell line. ARO cells, derived from a thyroid anaplastic carcinoma, were exposed to different concentrations of the drug. Liposomes formulations were made up of 1,2 dipalmitoyl-sn-glycero-3-phosphocholine/cholesterol/1,2-distearoyl-sn-glycero-3 phosphoethanolamine-MPEG (8:3:1 molar ratio). Cell viability was assessed by both trypan bleu dye exclusion assay and fluorimetric analysis of cell DNA content. RESULTS: A cytotoxic effect of free gemcitabine was present only after 72 h incubation (ARO cell mortality increased of approximately 4 fold over control at 1 microM, 7 fold at 100 microM). When gemcitabine was encapsulated in liposomes, a significant effect was observed by using lower concentrations of the drug (increased cell mortality of 2.4 fold vs. control at 0.3 microM) and earlier exposure time (24 h). CONCLUSION: These findings show that, in vitro against human thyroid cancer cells, the gemcitabine incorporation within liposomes enhances the drug cytotoxic effect with respect to free gemcitabine, thus suggesting a more effective drug uptake inside the cells. This may allow the use of new formulations with lower dosages (side effect free) for the treatment of anaplastic human thyroid tumours. PMID- 15363095 TI - Tissue detection of natural killer cells in colorectal adenocarcinoma. AB - BACKGROUND: Natural killer (NK) cells represent a first line of defence against a developing cancer; however, their exact role in colorectal cancer remains undetermined. The aim of the present study was to evaluate the expression of CD16 and CD57 [immunohistochemical markers of natural NK cells] in colorectal adenocarcinoma. METHODS: Presence of NK cells was investigated in 82 colorectal adenocarcinomas. Immunohistochemical analysis was performed, using 2 monoclonal antibodies (anti-Fc Gamma Receptor II, CD16 and an equivalent to Leu-7, specific for CD-57). The number of immunopositive cells (%) was evaluated by image analysis. The cases were characterized according to: patient gender and age, tumor location, size, grade, bowel wall invasion, lymph node metastases and Dukes' stage. RESULTS: NK cells were detected in 79/82 cases at the primary tumor site, 27/33 metastatic lymph nodes and 3/4 hepatic metastases; they were detected in levels similar to those reported in the literature, but their presence was not correlated to the clinical or pathological characteristics of the series, except for a negative association with the patients' age (p = 0.031). CONCLUSIONS: Our data do not support an association of NK cell tissue presence with clinical or pathological variables of colorectal adenocarcinoma, except for a negative association with the patients' age; this might possibly be attributed to decreased adhesion molecule expression in older ages. PMID- 15363096 TI - An integrated 4249 marker FISH/RH map of the canine genome. AB - BACKGROUND: The 156 breeds of dog recognized by the American Kennel Club offer a unique opportunity to map genes important in genetic variation. Each breed features a defining constellation of morphological and behavioral traits, often generated by deliberate crossing of closely related individuals, leading to a high rate of genetic disease in many breeds. Understanding the genetic basis of both phenotypic variation and disease susceptibility in the dog provides new ways in which to dissect the genetics of human health and biology. RESULTS: To facilitate both genetic mapping and cloning efforts, we have constructed an integrated canine genome map that is both dense and accurate. The resulting resource encompasses 4249 markers, and was constructed using the RHDF5000-2 whole genome radiation hybrid panel. The radiation hybrid (RH) map features a density of one marker every 900 Kb and contains 1760 bacterial artificial chromosome clones (BACs) localized to 1423 unique positions, 851 of which have also been mapped by fluorescence in situ hybridization (FISH). The two data sets show excellent concordance. Excluding the Y chromosome, the map features an RH/FISH mapped BAC every 3.5 Mb and an RH mapped BAC-end, on average, every 2 Mb. For 2233 markers, the orthologous human genes have been established, allowing the identification of 79 conserved segments (CS) between the dog and human genomes, dramatically extending the length of most previously described CS. CONCLUSIONS: These results provide a necessary resource for the canine genome mapping community to undertake positional cloning experiments and provide new insights into the comparative canine-human genome maps. PMID- 15363097 TI - Synchronously diagnosed pre-sacral neurofibroma and cutaneous spitzoid melanoma: a fortuitous association? AB - BACKGROUND: At a U.S prevalence of 1 in 3000, Neurofibromatosis type-1 (NF-1) is a relatively common disorder. Amongst a variety of others, occurrence of 2 or more neurofibromas in the same patient represents one of the major diagnostic criteria for this disorder. Rarely, ocular, cutaneous or anorectal malignant melanomas may be identified in patients with NF-1, This rare association has caused controversy as to whether patients with NF-1 have an inherently higher risk for melanomas or whether the associations can be explained by chance alone. CASE PRESENTATION: The purpose of this report is to highlight the unusual confluence of rare clinicopathologic features in a patient without NF-1. The patient was diagnosed with an 8.5 cm pre-sacral neurofibroma and was shortly thereafter diagnosed with a cutaneous malignant melanoma showing spitzoid features. Pre-sacral neurofibromas are rare in patients without NF-1; likewise, malignant spitzoid melanoma, a controversial histopathological entity, is distinctly uncommon. CONCLUSIONS: The synchronous diagnoses of these neural crest derived tumor entities in a patient without neurofibromatosis lends credence to the view that when these two lesions occur in patients with NF-1, the association is coincidental. PMID- 15363098 TI - Anogenital distance in human male and female newborns: a descriptive, cross sectional study. AB - BACKGROUND: In animal studies of the effects of hormonally active agents, measurement of anogenital distance (AGD) is now routine, and serves as a bioassay of fetal androgen action. Although measurement of AGD in humans has been discussed in the literature, to our knowledge it has been measured formally in only two descriptive studies of females. Because AGD has been an easy-to-measure, sensitive outcome in animals studies, we developed and implemented an anthropometric protocol for measurement of AGD in human males as well as females. METHODS: We first evaluated the reliability of the AGD measures in 20 subjects. Then measurements were taken on an additional 87 newborns (42 females, 45 males). All subjects were from Morelos, Mexico. RESULTS: The reliability (Pearson r) of the AGD measure was, for females 0.50, and for males, 0.64. The between-subject variation in AGD, however, was much greater than the variation due to measurement error. The AGD measure was about two-fold greater in males (mean, 22 mm) than in females (mean, 11 mm), and there was little overlap in the distributions for males and females. CONCLUSION: The sexual dimorphism of AGD in humans comprises prima facie evidence that this outcome may respond to in utero exposure to hormonally active agents. PMID- 15363099 TI - Food and nutrient intake in relation to mental wellbeing. AB - BACKGROUND: We studied food consumption and nutrient intake in subjects with depressed mood, anxiety and insomnia as indices of compromised mental wellbeing. METHODS: The study population consisted of 29,133 male smokers aged 50 to 69 years who entered the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study in 1985-1988. This was a placebo-controlled trial to test whether supplementation with alpha-tocopherol or beta-carotene prevents lung cancer. At baseline 27,111 men completed a diet history questionnaire from which food and alcohol consumption and nutrient intake were calculated. The questionnaire on background and medical history included three symptoms on mental wellbeing, anxiety, depression and insomnia experienced in the past four months. RESULTS: Energy intake was higher in men who reported anxiety or depressed mood, and those reporting any such symptoms consumed more alcohol. Subjects reporting anxiety or depressed mood had higher intake of omega-3 fatty acids and omega-6 fatty acids. CONCLUSIONS: Our findings conflict with the previous reports of beneficial effects of omega-3 fatty acids on mood. PMID- 15363100 TI - Validity and reproducibility of an interviewer-administered food frequency questionnaire for healthy French-Canadian men and women. AB - OBJECTIVE: To evaluate the validity (study 1) and the reproducibility (study 2) of an interviewer-administered food frequency questionnaire (FFQ). METHOD: The FFQ was designed at Laval University and contains 91 items and 33 subquestions. STUDY 1: The FFQ was compared against a 3-day food record (2 week-days and 1 weekend-day), at week 0, 6 and 12 of a nutritional intervention. STUDY 2: In order to evaluate the reproducibility of the FFQ, 2 registered dietitians administered the FFQ 4-weeks apart among subjects who were not part of the nutritional intervention. RESULTS: STUDY 1: Mean values for intake of most nutrients assessed by the FFQ and by the 3-day food record were not statistically different. Energy-adjusted correlation coefficients for major macronutrients ranged from 0.36 for proteins to 0.60 for carbohydrates (p < or = 0.01). Agreement analysis revealed that on average, 35% of the subjects were classified in the same quartile when nutrients were assessed by either the 3-day food record or the FFQ. STUDY 2: Significant associations were observed between dietary measurements derived from the two FFQs administered 4 weeks apart. Correlation coefficients for the reproducibility of macronutrients ranged from 0.66 for carbohydrates to 0.83 for lipids after energy adjustment. On average, 46% of the subjects were classified in the same quartile when nutrient intakes were assessed by either FFQ. CONCLUSION: These data indicated that the FFQ developed has a good validity and is reproducible. PMID- 15363101 TI - Nutritional care of Danish medical inpatients: effect on dietary intake and the occupational groups' perspectives of intervention. AB - BACKGROUND: Many patients do not eat and drink sufficiently during hospitalisation. The clinical consequences of this under nutrition include lassitude, an increased risk of complications and prolonged convalescence. The aim of the study was 1) to introduce intervention targeting nutritional care for medical inpatients, 2) to investigate the effect of this intervention, and 3) to investigate the occupational groups' attitudes towards nutritional intervention and nutritional care in general. METHODS: The design was to determinate the extent to which the protein and energy requirements of medical inpatients were met before and after intervention. Dietary protein and energy intakes were assessed by 72-hour weighed food records. A total number of 108 medical patients at four bed sections and occupational groups in the two intervention bed sections, Aarhus University Hospital, Denmark participated. The intervention included introduction and implementation of nursing procedures targeting nutritional care during a five-month investigation period using standard food produced at the hospital. The effect of intervention for independent groups of patients were tested by one-way analysis of variance. After the intervention occupational groups were interviewed in focus groups. RESULTS: Before the intervention hospital food on average met 72% of the patients' protein requirement and 85% of their energy requirement. After intervention hospital food satisfied 85% of the protein and 103% of the energy requirements of 14 patients in one intervention section and 56% of the protein and 76% of the energy requirement of 17 patients in the other intervention section. Hospital food satisfied 61% of the protein and 75% of the energy requirement in a total of 29 controls. From the occupational groups' point of view lack of time, lack of access to food, and lack of knowledge of nutritional care for patients were identified as barriers to better integration of nutritional care into the overall care provision. CONCLUSION: There was ample room for improving the extent to which standard hospital food satisfies patients' protein and energy requirements, but implementation of procedures addressing nutritional care were difficult, especially at bed sections with a large staff turnover. PMID- 15363102 TI - Additional collection devices used in conjunction with the SurePath Liquid-Based Pap Test broom device do not enhance diagnostic utility. AB - BACKGROUND: We have previously shown that use of an EC brush device in combination with the Rovers Cervex-Brush (SurePath broom) offered no significant improvement in EC recovery. Here we determine if use of additional collection devices enhance the diagnostic utility of the SurePath Pap for gynecologic cytology. METHODS: After informed consent, 37 women ages 18-56 receiving their routine cervical examinations were randomized into four experimental groups. Each group was first sampled with the SurePath broom then immediately re-sampled with an additional collection device or devices. Group 1: Rover endocervix brush (n = 8). Group 2: Medscand CytoBrush Plus GT (n = 7). Group 3: Rover spatula + endocervix brush (n = 11). Group 4: Medscand spatula + CytoBrush Plus GT (n = 11). RESULTS: Examination of SurePath broom-collected cytology yielded the following abnormal diagnoses: atypia (n = 2), LSIL (n = 5) and HSIL (n = 3). Comparison of these diagnoses to those obtained from paired samples using the additional collection devices showed that use of a second and or third device yielded no additional abnormal diagnoses. Importantly, use of additional devices did not improve upon the abnormal cell recovery of the SurePath broom and in 4/10 cases under-predicted or did not detect the SurePath broom-collected lesion as confirmed by cervical biopsy. Finally, in 36/37 cases, the SurePath broom successfully recovered ECs. Use of additional devices, in Group 3, augmented EC recovery to 37/37. CONCLUSIONS: Use of additional collection devices in conjunction with the SurePath broom did not enhance diagnostic utility of the SurePath Pap. A potential but not significant improvement in EC recovery might be seen with the use of three devices. PMID- 15363103 TI - Radiation enteropathy and leucocyte-endothelial cell reactions in a refined small bowel model. AB - BACKGROUND: Leucocyte recruitment and inflammation are key features of high dose radiation-induced tissue injury. The inflammatory response in the gut may be more pronounced following radiotherapy due to its high bacterial load in comparison to the response in other organs. We designed a model to enable us to study the effects of radiation on leucocyte-endothelium interactions and on intestinal microflora in the murine ileum. This model enables us to study specifically the local effects of radiation therapy. METHOD: A midline laparotomy was performed in male C57/Bl6 mice and a five-centimetre segment of ileum is irradiated using the chamber. Leucocyte responses (rolling and adhesion) were then analysed in ileal venules 2 - 48 hours after high dose irradiation, made possible by an inverted approach using intravital fluorescence microscopy. Furthermore, intestinal microflora, myeloperoxidase (MPO) and cell histology were analysed. RESULTS: The highest and most reproducible increase in leucocyte rolling was exhibited 2 hours after high dose irradiation whereas leucocyte adhesion was greatest after 16 hours. Radiation reduced the intestinal microflora count compared to sham animals with a significant decrease in the aerobic count after 2 hours of radiation. Further, the total aerobic counts, Enterobacteriaceae and Lactobacillus decreased significantly after 16 hours. In the radiation groups, the bacterial count showed a progressive increase from 2 to 24 hours after radiation. CONCLUSION: This study presents a refinement of a previous method of examining mechanisms of radiation enteropathy, and a new approach at investigating radiation induced leucocyte responses in the ileal microcirculation. Radiation induced maximum leucocyte rolling at 2 hours and adhesion peaked at 16 hours. It also reduces the microflora count, which then starts to increase steadily afterwards. This model may be instrumental in developing strategies against pathological recruitment of leucocytes and changes in intestinal microflora in the small bowel after radiotherapy. PMID- 15363104 TI - Scoliosis treatment using a combination of manipulative and rehabilitative therapy: a retrospective case series. AB - BACKGROUND: The combination of spinal manipulation and various physiotherapeutic procedures used to correct the curvatures associated with scoliosis have been largely unsuccessful. Typically, the goals of these procedures are often to relax, strengthen, or stretch musculotendinous and/or ligamentous structures. In this study, we investigate the possible benefits of combining spinal manipulation, positional traction, and neuromuscular reeducation in the treatment of idiopathic scoliosis. METHODS: A total of 22 patient files were selected to participate in the protocol. Of these, 19 met the study criterion required for analysis of treatment benefits. Anteroposterior radiographs were taken of each subject prior to treatment intervention and 4-6 weeks following the intervention. A Cobb angle was drawn and analyzed on each radiograph, so pre and post comparisons could be made. RESULTS: After 4-6 weeks of treatment, the treatment group averaged a 17 degrees reduction in their Cobb angle measurements. None of the patients' Cobb angles increased. A total of 3 subjects were dismissed from the study for noncompliance relating to home care instructions, leaving 19 subjects to be evaluated post-intervention. CONCLUSIONS: The combined use of spinal manipulation and postural therapy appeared to significantly reduce the severity of the Cobb angle in all 19 subjects. These results warrant further testing of this protocol. PMID- 15363105 TI - Elevated midbrain serotonin transporter availability in mixed mania: a case report. AB - BACKGROUND: Results obtained from brain imaging studies indicate that serotonin transporter (SERT) and dopamine transporter (DAT) densities are altered in major depression. However, no such studies have been published on current mania or hypomania. CASE PRESENTATION: In this single photon emission computed tomography (SPECT) study with [123I]nor-beta-CIT we present a case with simultaneous symptoms of major depression and hypomania. She had an elevated serotonin transporter availability (SERT) in the midbrain and elevated dopamine transporter availability (DAT) in the striatum, which normalised in a one-year follow-up period during which she received eight months of psychodynamic psychotherapy. CONCLUSIONS: To our knowledge, this is the first report on SERT and DAT associated with mania. In our case the availability of both SERT in the midbrain and DAT in the striatum were elevated at baseline and declined during psychotherapy, while the SERT and DAT of the depressed controls increased during psychotherapy. Symptoms of hypomania in the case were alleviated during psychotherapy. Clinical recovery was also reflected in the Hamilton Depression Rating Scale (HDRS) scores. PMID- 15363106 TI - High-dose ibuprofen therapy associated with esophageal ulceration after pneumonectomy in a patient with cystic fibrosis: a case report. AB - BACKGROUND: Lung disease in patients with cystic fibrosis is thought to develop as a result of airway inflammation, infection, and obstruction. Pulmonary therapies for cystic fibrosis that reduce airway inflammation include corticosteroids, rhDNase, antibiotics, and high-dose ibuprofen. Despite evidence that high-dose ibuprofen slows the progression of lung disease in patients with cystic fibrosis, many clinicians have chosen not to use this therapy because of concerns regarding potential side effects, especially gastrointestinal bleeding. However, studies have shown a low incidence of gastrointestinal ulceration and bleeding in patients with cystic fibrosis who have been treated with high-dose ibuprofen. CASE PRESENTATION: The described case illustrates a life-threatening upper gastrointestinal bleed that may have resulted from high-dose ibuprofen therapy in a patient with CF who had undergone a pneumonectomy. Mediastinal shift post-pneumonectomy distorted the patient's esophageal anatomy and may have caused decreased esophageal motility, which led to prolonged contact of the ibuprofen with the esophagus. The concentrated effect of the ibuprofen, as well as its systemic effects, probably contributed to the occurrence of the bleed in this patient. CONCLUSIONS: This report demonstrates that gastrointestinal tract anatomical abnormalities or dysmotility may be contraindications for therapy with high-dose ibuprofen in patients with cystic fibrosis. PMID- 15363107 TI - Evaluation of the prophylactic use of mitomycin-C to inhibit haze formation after photorefractive keratectomy in high myopia: a prospective clinical study. AB - BACKGROUND: To study the effect of prophylactic application of mitomycin-C on haze formation in photorefractive keratectomy (PRK) for high myopia. METHODS: Fifty-four eyes of 28 myopic patients were enrolled in this prospective study. All eyes were operated by PRK followed by 0.02% mitomycin-C application for two minutes and washed with 20 ml normal saline afterwards. All eyes were examined thoroughly on the first 7 days and one month after surgery; 48 eyes (88.9%) at 3 and 6 months postoperatively. Hanna grading (in the scale of 0 to 4+) was used for assessment of corneal haze. RESULTS: The mean spherical equivalent refraction (SE) was -7.08 diopters (D) +/- 1.11 (SD) preoperatively. Six months after surgery, 37 eyes (77.1%) achieved an uncorrected visual acuity (UCVA) of 20/20 or better, all eyes had a UCVA of 20/40 or better and 45 (93.7%) eyes had an SE within +/- 1.00D. One month postoperatively, 2 eyes (3.7%) had grade 0.5+ of haze, while at 3 and 6 months after surgery no visited eye had haze at all. All eyes had a best corrected visual acuity (BCVA) of 20/40 or better and there were no lost lines in BCVA by 6 months after surgery. In spatial frequencies of 6 and 12 cycles per degree contrast sensitivity had decreased immediately after PRK and it had increased 1.5 lines by the 6th postoperative month compared to the preoperative data. CONCLUSIONS: The results show the efficacy of mitomycin-C in preventing corneal haze after treatment of high myopia with PRK. This method- PRK + mitomycin-C - can be considered an alternative treatment for myopic patients whose corneal thicknesses are inadequate for laser in situ keratomileusis (LASIK). However, the results should be confirmed in longer follow-ups. PMID- 15363108 TI - Transformation and scattering activities of the receptor tyrosine kinase RON/Stk in rodent fibroblasts and lack of regulation by the jaagsiekte sheep retrovirus receptor, Hyal2. AB - BACKGROUND: The envelope (Env) protein of jaagsiekte sheep retrovirus (JSRV) can transform cells in culture and is likely to be the main factor responsible for lung cancer induction by JSRV in animals. A recent report indicates that the epithelial-cell transforming activity of JSRV Env depends on activation of the cell-surface receptor tyrosine kinase Mst1r (called RON for the human and Stk for the rodent orthologs). In the immortalized line of human epithelial cells used (BEAS-2B cells), the virus receptor Hyal2 was found to bind to and suppress the activity of RON. When Env was expressed it bound to Hyal2 causing its degradation, release of RON activity from Hyal2 suppression, and activation of pathways resulting in cell transformation. METHODS: Due to difficulty with reproducibility of the transformation assay in BEAS-2B cells, we have used more tractable rodent fibroblast models to further study Hyal2 modulation of RON/Stk transforming activity and potential effects of Hyal2 on RON/Stk activation by its natural ligand, macrophage stimulating protein (MSP). RESULTS: We did not detect transformation of NIH 3T3 cells by plasmids expressing RON or Stk, but did detect transformation of 208F rat fibroblasts by these plasmids at a very low rate. We were able to isolate 208F cell clones that expressed RON or Stk and that showed changes in morphology indicative of transformation. The parental 208F cells did not respond to MSP but 208F cells expressing RON or Stk showed obvious increases in scattering/transformation in response to MSP. Human Hyal2 had no effect on the basal or MSP-induced phenotypes of RON-expressing 208F cells, and human, mouse or rat Hyal2 had no effect on the basal or MSP-induced phenotypes of Stk-expressing 208F cells. CONCLUSIONS: We have shown that RON or Stk expression in 208F rat fibroblasts results in a transformed phenotype that is enhanced by addition of the natural ligand for these proteins, MSP. Hyal2 does not directly modulate the basal or MSP-induced RON/Stk activity, although it is possible that adaptor proteins might mediate such signaling in other cell types. PMID- 15363109 TI - Identification of the 15FRFG domain in HIV-1 Gag p6 essential for Vpr packaging into the virion. AB - The auxiliary regulatory protein Vpr of HIV-1 is packaged in the virion through interaction with the Gag C-terminal p6 domain. Virion packaging of Vpr is critical for Vpr to exert functions in the HIV-1 life cycle. Previous studies suggest that Vpr interacts with a (Lxx)4 domain in p6 for virion packaging. In the present study, mutational analysis of HIV-1 Gag p6 domain was performed in the context of the HIV-1 genome to examine the effect on virion packaging of Vpr. Surprisingly, Ala substitutions for Leu44 and Phe45 in the (Lxx)4 domain or deletion of the whole (Lxx)4 domain (amino acid #35-52 of the Gag p6 domain) did not affect Vpr virion packaging. Vpr virion packaging was normal when amino acid #1-23 of the Gag p6 domain was preserved. Most importantly, Ala substitutions for Phe15, Arg16 and Phe17 in the context of amino acid #1-23 of the Gag p6 domain abolished Vpr virion packaging. Single Ala substitutions for Phe15 and Phe17 also abolished Vpr virion packaging, whereas Ala substitution for Arg16 had no effect. Our studies have revealed a novel signal sequence for Vpr packaging into the HIV 1 virion. The 15FRFG domain in p6 resembles the FxFG repeat sequences commonly found in proteins of the nuclear pore complex. These results have provided novel insights into the process of virion packaging of Vpr and suggest for the first time that Vpr may recognize the FxFG domain for both virion packaging and association with nuclear pores. PMID- 15363110 TI - Ambiguous allele combinations in HLA Class I and Class II sequence-based typing: when precise nucleotide sequencing leads to imprecise allele identification. AB - Sequence-based typing (SBT) is one of the most comprehensive methods utilized for HLA typing. However, one of the inherent problems with this typing method is the interpretation of ambiguous allele combinations which occur when two or more different allele combinations produce identical sequences. The purpose of this study is to investigate the probability of this occurrence. We performed HLA-A,-B SBT for Exons 2 and 3 on 676 donors. Samples were analyzed with a capillary sequencer. The racial distribution of the donors was as follows: 615-Caucasian, 13-Asian, 23-African American, 17-Hispanic and 8-Unknown. 672 donors were analyzed for HLA-A locus ambiguities and 666 donors were analyzed for HLA-B locus ambiguities. At the HLA-A locus a total of 548 total ambiguous allele combinations were identified (548/1344 = 41%). Most (278/548 = 51%) of these ambiguities were due to the fact that Exon 4 analysis was not performed. At the HLA-B locus 322 total ambiguous allele combinations were found (322/1332 = 24%). The HLA-B*07/08/15/27/35/44 antigens, common in Caucasians, produced a large portion of the ambiguities (279/322 = 87%). A large portion of HLA-A and B ambiguous allele combinations can be addressed by utilizing a group-specific primary amplification approach to produce an unambiguous homozygous sequence. Therefore, although the prevalence of ambiguous allele combinations is high, if the resolution of these ambiguities is clinically warranted, methods exist to compensate for this problem. PMID- 15363111 TI - Skipping the co-expression problem: the new 2A "CHYSEL" technology. AB - The rapid progress in the field of genomics is increasing our knowledge of multi gene diseases. However, any realistic hope of gene therapy treatment for those diseases needs first to address the problem of co-ordinately co-expressing several transgenes. Currently, the use of internal ribosomal entry sites (IRESs) is the strategy chosen by many researchers to ensure co-expression. The large sizes of the IRESs (~0.5 kb), and the difficulties of ensuring a well-balanced co expression, have prompted several researchers to imitate a co-expression strategy used by many viruses: to express several proteins as a polyprotein. A small peptide of 18 amino acids (2A) from the foot-and-mouth disease virus (FMDV) is being used to avoid the need of proteinases to process the polyprotein. FMDV 2A is introduced as a linker between two proteins to allow autonomous intra ribosomal self-processing of polyproteins. Recent reports have shown that this sequence is compatible with different sub-cellular targeting signals and can be used to co-express up to four proteins from a single retroviral vector. This short peptide provides a tool to allow the co-expression of multiple proteins from a single vector, a useful technology for those working with heteromultimeric proteins, biochemical pathways or combined/synergistic phenomena. PMID- 15363112 TI - Prevention of non-communicable diseases in Pakistan: an integrated partnership based model. AB - Development and implementation of non-communicable disease (NCD) prevention polices in the developing countries is a multidimensional challenge. This article highlights the evolution of a strategic approach in Pakistan. The model is evidence-based and encompasses a concerted and integrated approach to NCDs. It has been modelled to impact a set of indicators through the combination of a range of actions capitalizing on the strengths of a public-private partnership. The paper highlights the merits and limitations of this approach. The experience outlines a number of clear imperatives for fostering an enabling environment for integrated NCD prevention public health models, which involve roles played by a range of stakeholders. It also highlights the value that such partnership arrangements bring in facilitating the mission and mandates of ministries of health, international agencies with global health mandates, and the non-profit private sector. The experience is of relevance to developing countries that have NCD programs running and those that need to develop them. It provides an empirical basis for enhancing the performance of the health system by fostering partnerships within integrated evidence-based models and permits an analysis of health systems models built on shared responsibility for the purpose of providing sustainable health outcomes. PMID- 15363113 TI - Just what does responsible board governance mean today? PMID- 15363114 TI - Fruit and vegetable consumption: benefits and barriers. AB - Few people on Prince Edward Island meet the goal of consuming five or more servings of vegetables and fruit a day. The main objective of this qualitative study was to explore the perceptions of the nutritional benefits and barriers to vegetable and fruit intake among adult women in Prince Edward Island. Participants were 40 women aged 20-49, with or without children at home, who were or were not currently meeting the objective of eating five or more fruit and vegetable servings a day. In-home, one-on-one interviews were used for data collection. Thematic analysis was conducted on the transcribed interviews. Data were examined for trustworthiness in the context of credibility, transferability, and dependability. Most participants identified one or more benefits of eating fruit and vegetables; however, comments tended to be non-specific. The main barriers that participants identified were effort, lack of knowledge, sociopsychological and socioenvironmental factors, and availability. Internal influences, life events, and food rules were identified as encouraging women to include vegetables and fruit in their diets. Given the challenges of effecting meaningful dietary change, dietitians must look for broader dietary behavioural interventions that are sensitive to women's perceptions of benefits and barriers to fruit and vegetable intake. PMID- 15363115 TI - What do Quebec children eat during their first six months? AB - Study objectives were to describe the introduction of foods and beverages and the use of vitamin and/or mineral supplements among infants aged six months, and to identify factors associated with following Quebec recommendations on introducing foods and beverages. Study subjects were primiparous women interviewed by phone when their infants were approximately six months old. Data were gathered on foods and beverages offered to infants, the age at which these were introduced, and supplements. The use of each food, beverage, and supplement was evaluated according to recommendations. Mean age at which foods were introduced and proportions of women who followed recommendations were calculated. Multivariate logistic regression models were used to identify the factors associated with following recommendations. Results indicated that among the 1,937 interviewed mothers, 212 (10.9%) exclusively breast-fed their infants before age four months, 1,073 (55.4%) breast-fed their infants at birth but gave them formula or foods before age four months, and 652 (33.7%) formula-fed their infants at birth. Factors associated with adherence to recommendations were breast-feeding exclusively, being older than 30 years, an annual family income of at least 60,000 dollars, and being a non-smoker. Few mothers followed all the recommendations. PMID- 15363116 TI - Nutrition services in Canadian neonatal follow-up programs. AB - The benefits of nutrition assessment and support of the high-risk infant are well established. The premature infant remains vulnerable for poor growth and developmental disabilities, thus requiring consistent monitoring, intervention, and follow-up care. The purpose of this study was to determine the registered dietitian's role in neonatal/perinatal follow-up programs. A survey was sent to the 26 follow-up programs in Canada. The questionnaire response rate was 81%. Registered dietitians were involved in 67% of these programs. Of these dietitians, 43% were assigned to neonatal/perinatal follow-up programs while 57% were involved only by consult. The average time that assigned registered dietitians devoted to programs was 0.35 full-time equivalents. Over 80% of the dietitians did ongoing development, evaluation, and modification of nutrition care plans; 71% screened new patients for nutritional risk, and 100% instructed patient families and developed teaching materials. The study findings will assist program planners who wish to establish a dietitian position in a neonatal/ perinatal follow-up program. For registered dietitians already working in such programs, the results may provide some guidance on role definition and expansion. PMID- 15363117 TI - Energy requirements of children with cerebral palsy. AB - Energy requirements of children and adolescents with cerebral palsy appear to be disease-specific and different from the current recommendations for healthy children, varying depending upon functional capacity, degree of mobility, severity of disease, and level of altered metabolism. Feeding problems are prevalent in many of these children, and can result in inadequate energy intake. Wasting of voluntary muscles, a common symptom of cerebral palsy, contributes to reduced resting energy needs; nevertheless, the location of the central nervous system lesion may also influence energy requirements. To guarantee individualized, accurate, and optimal energy recommendations for this population, resting energy expenditure should preferentially be measured by indirect calorimetry. Equations and formulae to predict healthy people's resting energy expenditure are available, but tend to overestimate these children's energy needs. Future studies should address the role of the central nervous system in regulating energy metabolism in this population. When adequately nourished, children and adolescents with cerebral palsy appear more tranquil and require decreased feeding time, which gives caregivers time to develop the child's functional independence and character. Understanding energy requirements of this population will provide caregivers and health professionals with guidelines for providing optimal nutritional status. PMID- 15363118 TI - A new approach to control of anemia in "at risk" infants and children around the world. 2004 Ryley-Jeffs memorial lecture. AB - An estimated 60-80% of the world's population is affected by iron deficiency. It is the most common preventable nutritional deficiency in the world, despite global goals for its reduction. Young children are the highest risk group, particularly during their rapid period of growth. Anemia in infants and young children is known to have a negative impact on motor and socioemotional development and cognitive function. A new intervention, microencapsulated ferrous fumarate Sprinkles, has been developed to treat iron deficiency. The development process took the intervention from the idea stage to the research stage to the implementation stage. Multiple micronutrient Sprinkles are a breakthrough in meeting the global challenge to reduce childhood anemia due to iron and other micronutrient deficiencies. The goal of the Sprinkles project is to reduce the global burden of childhood anemia by promoting healthy weaning practices, by advocating home-fortification strategies to increase the micronutrient content of weaning foods, and by making Sprinkles widely available, particularly in developing countries. PMID- 15363119 TI - [Influence of different conditions on two-dimensional gel electrophoresis of proteins from human acute promyelocytic leukemia cell line NB4]. AB - The aim of this study was to explore the effect of different conditions on two dimensional gel electrophoresis of proteins from human acute promyelocytic leukemia cell line NB4. The 24 cm pH 3-10 linear immobilized pH gradient (IPG) strips were chosen, the isoelectric focusing was carried out by using IPGphor. Then, the second-dimensional SDS-PAGE was performed. After silver staining, the gel was analyzed by ImageMaster 2D Elite. The results showed that low ion intensity sample washing buffer improved the performance of isoelectric focusing. The lysis buffer containing 7 mol/L urea and 40 mmol/L DTT could solubilize the most proteins from NB4 cell line. The rehydration solution containing thiourea and urea increased the low molecular weight protein points to be resolved in the area of basic end. The reasonable sample load and Volt/hour of NB4 were about 100 micro g and 63 200 V/h for the 24 cm pH 3-10 IPG strips. It is concluded that the proteins from NB4 and similar cell line are complicated and affected by many factors, so that, it is very important to select the right methods for sample preparation and the conditions of two-dimensional gel electrophoresis. PMID- 15363120 TI - Coexistence of tetrasomy 8 and trisomy 8 in acute promyelocytic leukemia (AML-M3) with t(15;17)(q22;q12). AB - This study was purposed to characterize the first case of acute promyelocitic leukemia (AML-M(3a)) with t(15;17), trisomy 8 and tetrasomy 8, and explore its characteristics of morphology, cytogenetics, molecular biology, immunology and clinical features. Morphological changes of peripheral blood and bone marrow smears were observed under microscope. Chromosome specimen was prepared by 24 h short-term culture of bone marrow cell, RHG-banding technique was used for karyotypic analysis. PML-RARa fusion gene transcript was detected by nested reverse transcription-polymerase chain reaction (nested RT-PCR). Interphase fluorescence in situ hybridization (FISH) using chromosome 8 centromere specific probe were carried out to detect abnormal numbers of chromosome 8. Immunophenotypic analysis was performed by flow cytometry. The results showed that peripheral blood smear revealed 65% promyelocyte, and bone marrow aspirate was hypercellular with 72.4% promyelocyte, moderately basophilic cytoplasm with numerous azurophilic granules. Karyotype analysis demonstrated 48, XY, +8, +8, t(15;17)(q22;q12) [16]/47, XY, +8, t(15;17)(q22;q12) [3]/46, XY, t(15;17)(q22;q12) [1]. RT-PCR assay revealed PML-RARa fusion gene transcript (+). FISH showed that the percentages of cells exhibiting 1, 2, 3, 4, 5, 6 green fluorescence signals were 0.5, 7, 19, 55, 18 and 0.5, respectively. This confirmed the presence of tetrasomy 8 and trisomy 8 and also revealed a low percentage of a pentasomy 8 clone. Immunophenotypes of the blasts displayed that CD13 (96.2%), CD33 (55.9%), CYMPO (93.5%) were positive. All the lymphoid markers tested were negative. The patient survival time was just 10 days. It is concluded that tetrasomy 8 is secondary cytogenetic event after t(15;17) in this case. It may be a consequence of clonal evolution of trisomy 8. t(15;17) AML-M(3) with tetrasomy 8 heralds a poor prognosis. PMID- 15363121 TI - [Clinical studies on expressions of Fas and mdr-1 in acute leukemia and their correlations]. AB - To explore the Fas and mdr-1 expression and their correlation in multidrug resistance (MDR), Fas and mdr-1 expressions of bone marrow from 59 patients with newly diagnosed AL before therapy and after complete remission were detected by direct immunofluorescence with flow cytometry and semi-quantitative RT-PCR, respectively. The results showed that in newly diagnosed AL patients, Fas expression in AML was higher than in ALL (P < 0.05), mdr-1 expression in AML and ALL had no difference (P > 0.05), the expressions of Fas and mdr-1 had correlation (r = -0.282, P < 0.05) in AL; the results of simple-variable and multivariable COX survival factor model analysis suggested that Fas and mdr-1 expressions were prognostic factors for the effect of therapy and survival. Log rank test, comparing the groups of Fas(+) with Fas(-), mdr-1(+) with mdr-1(-), demonstrated that the CR rates and median remission time of every two groups had significant difference. It is concluded that in AL, Fas and mdr-1 expressions have high correlation with the effect of treatment, Fas expression probably is one of the favorable prognostic factors, mdr-1 is an unfavorable prognostic and less effective factor. PMID- 15363122 TI - [Establishment and characterization of leukemic cell line U937 stably expressing exogenous RbAp46]. AB - To establish leukemic cell lines stably transfected by RbAp46 gene, electroporation was performed after optimizing the transfection condition for suspended cells. Under conditions of low voltage and high capacitance, RbAp46 was transfected into U937 by electroporation. Individual clones selected with G418 for 3 weeks were isolated. The integration and the protein levels of the exogenous RbAp46 in transfectants were determined by PCR and Western blot analysis, respectively. The subclone expressing high level of RbAp46 was then established. Viability of transfected cells was assayed by trypan blue exclusion. Cell number was counted daily to determine the growth rate. The results showed that growth rate of U937 cell lines expressing exogenous RbAp46 was about 50% lower than that in control. It is concluded that leukemic cell lines stably expressing exogenous RbAp46 were established and overexpression of RbAp46 inhibits the growth of U937 leukemic cells. PMID- 15363123 TI - Mutation analysis of SHIP gene in acute leukemia. AB - The SH2 domain containing inositol 5'-phosphatase (SHIP) was initially described as a 145 kD protein phosphorylated on tyrosines upon growth factor and cytokine stimulation. SHIP is predominately expressed in hematopoietic cells, and is a crucial negative regulator in the development of hematopoietic cells. To evaluate the role of the SHIP gene in human leukemogenesis, expression and mutation of SHIP gene in bone marrow and/or peripheral blood from 32 patients with acute myeloid leukemia (AML), 9 patients with acute lymphoblastic leukemia (ALL), as well as human hematopoietic cell lines were analyzed by reverse transcription polymerase chain reaction (RT-PCR), single strand conformational polymorphism (SSCP) and sequencing. The RT-PCR showed that all samples expressed SHIP gene. Mutations of SHIP gene were detected in 7 out of 32 AML patients (22%) and one out of 9 ALL patients (12%). Interestingly, two missense mutations that had been observed in one AML patient at diagnosis disappeared after complete remission (CR). In addition, Akt phosphorylation was prolonged and increased following IL-3 stimulation in this patient sample. In conclusion, data of this study demonstrate the mutation of the SHIP gene in acute leukemia for the first time and suggest a possible role of the mutation of this gene in the development of acute leukemia. SHIP serves as a tumor suppressor by negatively regulating the PI3K/Akt signaling pathway in hematopoietic cells. PMID- 15363124 TI - [Effect of S-2-(3-aminopropylamino) ethyl phosphorothioic acid on apoptosis and proliferation inhibition of HL-60 cell line]. AB - To study the effects of S-2-(3-aminopropylamino) ethyl phosphorothioic acid (WR 2721, amifostine) on proliferation inhibition and apoptosis of HL-60 human leukemia cell line, the cell apoptosis rate of HL-60 was determined by annexin V/PI double staining method. Cell proliferation and chemotherapy sensitivity were analyzed with XTT assay, and the changes of cell cycle were observed through flow cytometry. The results showed that WR-2721 could significantly inhibit HL-60 cell proliferation. After treatment (30 min, 37 degrees C) with WR-2721, the sensitivity of HL-60 cells to VP16 was enhanced, and the IC(50) descended from 52.5 micro g/ml to 40.5 microg/ml. After 72 hours treatment of HL-60 cells with WR-2721, the early apoptotic cells (annexin V-FITC positive/PI negative) were increased from (5.5 +/- 1.9)% to (48.5 +/- 8.4)% (P < 0.001), late apoptotic cells (annexin V-FITC positive/PI positive) were increased from (1.2 +/- 0.5)% to (39.0 +/- 4.0)% (P < 0.001), and HL-60 cells were arrested in G(2)-M phase. In conclusion, WR-2721 treatment can enhance HL-60 cell chemotherapy sensitivity to VP16, inhibit proliferation, induce apoptosis and accumulation of cells in G(2)-M phase. PMID- 15363125 TI - [Construction and analysis of subtractive cDNA library associated with multidrug resistance of acute leukemia]. AB - The study was aimed to construct subtractive cDNA library associated with multidrug resistance (MDR) of acute leukemia for screening genes related to MDR in leukemia. The improved PCR-based subtractive hybridization was performed to clone differential genes between HL-60/VCR and HL-60 cell line. The mRNA of HL 60/VCR and HL-60 cell line were isolated. Then the mRNA of HL-60/VCR group was reversely transcribed into cDNA by Cap-Finder method, and the mRNA of HL-60 was reversely transcribed into cDNA by ordinary method to be marked by biotin for the hybridization next with HL-60/VCR cDNA. After hybridizing, filtrating through the sephacryl S-400 column, absorbing by the magnetic beads, and amplifying by PCR method, the fragments were cloned by T-A method and the cDNA library was constructed. Then the quality of cDNA library was identified by dot-blotting hybridization method. The results showed that after constriction, the library demonstrated its good quality. There was a high proportion of large fragments in this library. From small amount of samples a large amount of candidate fragments could be screened rapidly at once by dot-blotting hybridization. It is concluded that a differentially-expressed subtractive cDNA library in MDR of leukemia with high quality and larger fragments can be efficiently constructed by improving subtractive hybridization and selective PCR method. PMID- 15363126 TI - [Effects of anti- CXCR4 monoclonal antibody on adhesiveness of human acute myelocytic leukemia cell line HL-60 and expression of Bcl-2, Fas proteins]. AB - To study the importance of chemokine SDF-1 in surviving of acute myelocytic leukemia cells HL-60, the adhesion ability of HL-60 and expression of Bcl-2, Fas protein when SDF-1 activity was blocked by anti-CXCR4 monoclonal antibody (12G5) were compared with those detected before MAb incubation, in this experiment, HL 60 cell were cultured and co-cultured with normal marrow stromal cell. The adhesion rate was detected while the expression of Bcl-2 and Fas proteins were assayed by immunohistochemical technique when SDF-1 activity was inhibited. The results showed that cell adhesion rate of HL-60 decreased while the expression Bcl-2 decreased and Fas increased. It is concluded that inhibition of SDF-1 activity increases cell apoptosis and thus reduces life-span of leukemia cell at certain level. PMID- 15363127 TI - [CXCR4 expression in B-lineage acute lymphocyte leukemia and its significance]. AB - The study was aimed to explore the chemokine receptor CXCR4 expression on the B lineage acute lymphocyte leukemia (B-ALL) cells of various differentiation stages and its relationship with myeloid antigen expression. Flow cytometry was used to detect the CXCR4 expression by means of double-fluorescence labeling with CD19/SCC gating. The results demonstrated that 92.9% B-ALL patients were positively expressed CXCR4. The CD10, CD34 antigens were differently expressed in differentiation stages of B-ALL. The immunotypes of (1) CD10(-)/CD34(+), (2) CD10(+)/CD34(+), (3) CD10(+)/CD34(-), (4) CD10(-)/CD34(-) presented at various differential stages from premature to mature. The positive rate of CXCR4 were (27.60 +/- 15.25)%, (30.95 +/- 15.50)%, (55.62 +/- 18.37)% and (77.25 +/- 10.86)% from (1) to (4) respectively. The median fluorescence intensity (MFI) of CXCR4 expression were 46.69 +/- 15.06, 47.43 +/- 12.39, 79.28 +/- 24.71 and 132.92 +/- 88.09. CXCR4 expressions were not significantly different between the premature stages of CD10(-)/CD34(+) and CD10(+)/CD34(+) subtypes, but both were lower than the CXCR4 expression in CD10(+)/CD34(-) and CD10(-)/CD34(-) subtypes. The highest incidence of CXCR4 expression was found in CD10(-)/CD34(-) B-ALL. The average level of CXCR4 expression on B-ALL cell with positive myeloid antigen CD13 or/and CD33 (my(+)B-ALL) was (12.56 +/- 3.88)% of positive rate and 39.82 +/- 11.58 of MFI, both of which were less than the positive rate (37.57 +/- 11.59)% and the MFI (50.72 +/- 13.34) on B-ALL cells with negative myeloid antigen expression (mye(-)B-ALL). In conclusion, the CXCR4 expression is associated with differentiation level of B-ALL cells and down-regulated when co-expressed with myeloid antigens. PMID- 15363128 TI - [Change of Bcl-2, Bax proteins and mitochondrial membrane protein on nitric oxide induced apoptosis in HL-60 cells]. AB - To investigate the possible mechanisms of nitric oxide (NO)-induced apoptosis in leukemia cell line HL-60, HL-60 cells in vitro were incubated with sodium nitroprusside (SNP), the in situ cell apoptosis quantitatively was assayed by TdT mediated dUTP nick end labeling (TUNEL), the cell cycle DNA and proteins expression of Bcl-2, Bax, mitochondrial membrane protein (APO2.7) were analyzed by flow cytometry. The results showed that SNP induced HL-60 cell apoptosis in a dosage- and time-dependent manner. After exposure to SNP at the concentration of 1.0 mmol/L for 48 hours, the percentage of apoptosis HL-60 was (42.2 +/- 3.5)% for subG1 and (52.5 +/- 7.6)% for TUNEL respectively, and they are significantly higher than those in control and potassium ferricyanide (PFC) groups as same concentration. During the apoptosis process, it showed a decrease of Bcl-2 protein and an increase of Bax protein and mitochondrial membrane protein in HL 60 cell, proteins of Bcl-2, Bax and mitochondrial membrane were expressed in a dosage- and time-dependent manner too. In conclusion, during the process of SNP induced apoptosis in HL-60 cell, the expression of mitochondrial membrane protein was increased, Bcl-2 and Bax proteins may be important regulators. PMID- 15363129 TI - [Identification of acute leukemia-specific genes from leukemia recipient/sibling donor pairs by distinguishing study with oligonucleotide microarrays]. AB - To explore differentially expressed genes in leukemia gene expression profile and identify main related genes in acute leukemia, gene expression profiles were analyzed in bone marrow/leucopheresis peripheral blood stem cells samples from 9 acute leukemia patients and their sibling donors with the use of oligonucleotide microarrays. 163 reported leukemia-related genes were involved in the study. The oligonucleotide primers were designed, synthesized and spotted on the chemical material-coated-glass plates in array. The total RNAs were isolated from nine patients' bone marrow or leucopheresis peripheral blood cells and from nine their sibling donors peripheral blood stem cells treated by G-CSF, then collected by CS 3000 cell selection machine, and were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP. The mixed probes were then hybridized to the oligonucleotide microarray. The results showed that in four patient/donor pairs with B-ALL, 5 up-regulated (RIZ, STK-1, T-cell leukemia/lymphoma 1A, Cbp/p300, Op18) and 1 down-regulated genes (hematopoietic proteoglycan core protein) were identified; In five patient/donor pairs with AML-M(4) and AML-M(5), 6 up regulated (STAT5B, ligand p62 for the Lck SH2, CST3, LTC4S, myeloid leukemia factor 2 and epb72) and 1 down-regulated genes (CCR5) were identified. In conclusion, on the basis of distinguishing study of specific genetic related recipient/sibling donor pairs, screening leukemia-related genes with oligonucleotide microarrays, a set of 13 up-regulated or down-regulated genes among 163 leukemia-related genes has been identified. The result has further confirmed that above genes play critical role in the molecular mechanism of acute leukemia. PMID- 15363130 TI - [Study on karyotype of 306 cases of myelodysplastic syndrome]. AB - The purpose of this study was to explore the significance of abnormal karyotype in diagnosis and prognosis estimation of myelodysplastic syndrome (MDS). Chromosome analysis were performed in 306 cases of MDS using the short-term culture of bone marrow cell and G-banding technique, and in partial cases FISH technique was used for this analysis. 93 out of 306 cases were followed up. The results showed that 144 cases (47.1%) had clonal chromosome aberrations. The most common chromosomal aberrations included +8, translocation, complex or high complex karyotype, -7/7q-, 20q-/-20, trisomy 1 or partial trisomy 1, +11/+11q-, 9/9q-, +9/9q+, -Y, dup(1q), +21. The rate of abnormal karyotype in refractory anemia with erythroblasts (RAEB) and refractory anemia with erythroblasts transformation (RAEBT) were much higher than in refractory anemia (RA) and refractory anemia with sideroblasts (RAS) (P < 0.05). The rate of abnormal karyotype among those cases with mutagen contact history were higher than those in cases without mutagen contact history. The patients with abnormal karyotype had a mean survival time much shorter than patients with normal karyotype (P < 0.005) and had a higher risk transforming into acute leukemia (P < 0.05). The worst outcome was observed in those patients with a complex or high complex karyotype, -7/7q- and trisomy 11. In conclusion, MDS is highly heterogeneous disorders and karyotype analysis is helpful for its diagnosis, treatment selection and prognosis estimation. PMID- 15363131 TI - [Cytogenetic and clinical analysis of -7/7q- abnormalities in acute leukemia and myelodysplastic syndrome]. AB - The objective was to study the incidence and prognosis significance of -7/7q- abnormalities in acute leukemia and myelodysplastic syndrome. Conventional cytogenetic analysis of R-band was used to test -7/7q- chromosome abnormalities in 410 patients with acute leukemia (AL), in 71 cases of myelodysplastic syndrome (MDS) and in 36 cases of chronic myelogenous leukemia in accelerated phase (CML AP). The results showed that the incidences of -7/7q- abnormalities in AL, MDS and CML-AP patients were 4.88%, 9.86% and 8.33% respectively. The -7/7q- abnormalities could be found in acute myeloblastic leukemia (AML) and acute lymphocytic leukemia (ALL), incidences of which were 4.70% and 6.25% (P > 0.05) respectively. 9 cases had -7 or 7q- as the sole chromosome abnormalities, 22 cases showed other additional chromosome abnormalities: -X, -5, +8, t(3; 3), t(11;16) and t(2;11). Monosomy -7 and 7q- abnormality clone was found in one patient with MDS-RAEB, and the number of cells with -7 abnormality was greater than that of 7q- abnormality cells. Four patients acquired CR among 7 patients with ALL after chemotherapy, but 2 out of 13 patients with AML achieved CR while 6 out of 7 patients with MDS transformed into AL. No patients with CML-AP achieved CR. In conclusion, -7/7q- is a frequent aberration in hematologic malignancies as well as AML and ALL. The monosomy -7 and 7q-abnormalities were detected in the same patient. The patients with -7/7q- abnormalities show poor prognosis. PMID- 15363132 TI - [Expression of WT1 gene in patients with myelodysplastic syndrome and acute leukemia]. AB - To study the expression and significance of WT1 gene in patients with myelodysplastic syndrome (MDS) and acute leukemia (AL), RT-PCR was applied to monitor WT1 gene expression in 22 patients with MDS and in 69 patients with AL. The results showed that the positive rate of WT1 mRNA in MDS-RA and MDS-RAS was lower than that in MDS-RAEB and MDS-RAEB-t (10% versus 91.7%, P < 0.01). WT1 mRNA could be expressed in all subtype of AL, It was detected in 69% of newly diagnosed and relapsed patients, and in 12.5% patients CR. There was no difference at the relative expression level between newly diagnosed AL patients and relapsed patients, while the relative level of WT1 in MDS-RAEB and MDS-RAEB-t was lower than that in newly diagnosed AL. The CR rate in AML patients with positive expression was lower than that in patients with negative expression (41% versus 78%, P /= 1 had lower CR rate (18%) than those with relative level < 1 (55%). It is concluded that the expression of WT1 gene in patients with MDS-RAEB and RAEB-t was higher than that in patients with RA and RAS. The detection of WT1 gene may be useful for assessing disease progress of patients with MDS. The expression of WT1 gene and its expression level have associated with the prognosis of newly diagnosed patients with AL, that WT1 gene may be an independent prognostic factor in AML. PMID- 15363133 TI - [Effect of 5-aza-2'-deoxycytidine on cell of high-risk patients with myelodysplastic syndrome in vitro]. AB - To investigate the effect of 5-aza-2'-deoxycytidine (5-Aza-CdR) on cell of high risk patients with myelodysplastic syndrome (MDS) in vitro, the growth inhibition of MUTZ-1 cell induced by 5-Aza-CdR was detected by MTT method; apoptosis was detected by morphological observation and translocation of phosphatidylserine (PS) was examined by flow cytometry assay; the expressions of P15INK4B, DNA methyltransferases (DNMT)(1), DNMT(3A) and DNMT(3B) gene on mRNA level were detected by RT-PCR; methylation of p15INK4B gene in MUTZ-1 cells was detected by PCR using methylation specific primer (MSP). The results showed that 5-Aza-CdR inhibited the growth of MUTZ-1 cells. The IC(50) values of 24, 48 and 72 hours were 6.75, 2.82 and 5.45 mmol/L respectively. Characteristic changes of apoptosis emerged in MUTZ-1 cells after being exposed to 5-Aza-CdR in the different concentration from 0.8 mmol/L to 3.2 mmol/L, and the positive cells of annexin V on the membrane of MUTZ-1 cells were analyzed by flow cytometry. 5-Aza-CdR could activate the p15INK4B gene expression in MUTZ-1 cells by demethylation of the p15INK4B gene in a dose-dependent manner after the cells were treated for 48 hours. Furthermore, 5-Aza-CdR could significantly down-regulate the expressions of DNA methyltransferase genes DNMT(3A) at mRNA level in a dose dependent manner. However, it had no effects on DNMT(1) gene and DNMT(3B) gene. It is concluded that 5-Aza-CdR can inhibit the growth of MUTZ-1 cells and induce the apoptosis of these cells within the range of concentration from 0.8 mmol/L to 3.2 mmol/L, which may be one of the mechanisms of antitumor effects of 5-Aza-CdR. The drug can activate the expression of p15INK4B gene in MUTZ-1 cells by demethylation of the p15INK4B gene through inhibiting the expression of DNMT(3A) gene. It may be the mechanism of 5-Aza-CdR in the treatments of MDS. PMID- 15363134 TI - [Multiplex PCR for detecting genotypes of deletional alpha-thalassemia]. AB - To investigate the clinical application of multiplex PCR in detecting genotypes of deletional alpha-thalassemia in South China and observe the distribution frequency of alpha-globin gene deletion, 145 patients with silent carrier, alpha thalassemia trait or HbH were identified by M-PCR and 1.2% agarose gel electrophoresis. There are 1.3, 1.6, 1.8 and 2.0 kb bands which indicate --(SEA), -alpha(4.2), alphaalpha and -alpha(3.7), respectively. The results showed that among 145 patients, 100 patients with --(SEA)/alphaalpha (68.9%), 15 with alpha(3.7)/alphaalpha (10.3%), 8 with -alpha(4.2)/alphaalpha (5.52%), 2 with alpha(3.7)/-alpha(4.2) (1.38%), 1 with -alpha(3.7)/-alpha(3.7) (0.69%), 1 with alpha(4.2)/-alpha(4.2) (0.69%), 14 with --(SEA)/-alpha(3.7) (9.65%), 2 with - (SEA)/-alpha(4.2) (1.38%) were found. Two patients prenatal diagnosed were confirmed with Bart's hydrops fetuses. In conclusion, M-PCR analysis is a simple, rapid and accurate method for detection of alpha-thalassemia gene deletion. This technique is helpful in screening, carrier identification and prenatal diagnosis of deletional alpha-thalassemia. PMID- 15363135 TI - [Effects of normal human bone marrow fibroblastoid stromal cell line on the proliferation of multiple myeloma cell line RPMI8226]. AB - To investigate the effects of normal human bone marrow fibroblastoid stromal cell lines HFCL on the proliferation of multiple myeloma cell lines RPMI8226, the co culture system of RPMI8226 with HFCL was established, the adhesion ratio was determined by MTT colorimetric assay, growth curves were determined by trypan blue exclusion, the mitotic index (MI) of RPMI8226 cell was observed by Wright Giemsa staining. Flow cytometry and Western blot were used to study the changes of cell cycle and expression of proliferating cell nuclear antigen (PCNA), respectively. The results showed that in co-culture the adhesion ratio of RPMI8226 after 1 hour was 29.4%; the proliferation of RPMI8226 cell line in direct contact with HFCL cell line was inhibited as compared with RPMI8226 in suspension. No obvious changes were observed in RPMI8226 cell separated by transwell inserts. The percentage of G(1) phase cells of RPMI8226 cell line in direct contact with HFCL was higher than that of RPMI8226 in suspension, and the percentage of S phase cells was lower. Moreover, the MI of RPMI8226 cell line in suspension was higher than that of RPMI8226 cell line in direct contact with HFCL cell. The expression of PCNA in RPMI8226 cell line in suspension was higher than that of RPMI8226 cell in direct contact with HFCL cell. It is concluded that the normal human bone marrow fibroblastoid stromal cell HFCL inhibits the proliferation and progression of cell cycle of multiple myeloma cell line, RPMI8226. PMID- 15363136 TI - [Study on bone marrow megakaryocytes in children patients with idiopathic thrombocytopenic purpura]. AB - To observe the morphological characteristics and hematopoietic function of bone marrow megakaryocyte (MK) in children patients with idiopathic thrombocytopenic purpura (ITP), and to preliminary analyse the cause and mechanism of thrombocytopenia. CD41 McAb immunohistochemical technique was used to detect micromegakaryocyte in bone marrow smear. Plasma clot culture and CD41 McAb immunohistochemical technique were used for the MK-colony forming assay. The results showed that there was no statistical difference of the positive rate of micromegakaryocyte between groups of ITP and control, but type I lymphocyte-like micromegakaryocyte was infrequent. The number of micromegakaryocyte and the formation rates of CFU-MK and BFU-MK in ITP group were significantly higher than those in control group. The normal MK releasing platelet could be easily found in the culture system. The MK colony formation rate was decreased in a patient with chronic ITP. In conclusion, the increment of type II, III, IV micromegakaryocytes is one of pathologic phenomenon of ITP. These small megakaryocytes can develop and mature to normal megakaryocytes in the condition of ex vivo culture. The developmental abnormity of MK is a possible reason for thrombocytopenia among partial patients with ITP, especially the chronic cases. PMID- 15363137 TI - [Detection of minimal residual disease of chronic myeloid leukemia patients after allogeneic stem cell transplantation by combination of STR-PCR with RT-PCR]. AB - In order to evaluate relapse predication ability of STR-PCR combining with qualitative RT-PCR for the bar/abl transcripts to the patient with chronic myeloid leukemia (CML) fulfilled allogeneic stem cell transplantation (allo HSCT), 24 patients with CML after allo-HSCT were dynamically investigated for MRD, quantitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr/abl transcripts was detected by nested RT PCR. The results showed that persistent full donor chimerism (DC >/= 95%) was associated with an absence of MRD. All patients with stable MC (90% 0.05). It is concluded that this piezo-electric immunosensor detection has many advantages, such as convenient operation, real time operation and control, qualitative detection, and less cost. This method can be used for acute leukemia immunology classification. PMID- 15363143 TI - [Genotyping of ABO loci in para-Bombay type individuals]. AB - To study the molecular genetic basis of ABO alleles in para-Bombay type individuals, samples from five para-Bombay type individuals identified by serologic tests including absorption-elution tests, saliva neutralizing or inhibitor substances tests, were genotyped by using PCR-SSP based ABO genotyping. Exon 6 and exon 7 at the ABO locus for all 5 samples were sequenced. The results showed that the ABO genotypes of five para-Bombay samples were A102B1, A102B1, A102O1, A102B1, B1O1 respectively, the direct DNA sequencing results were in accordance with the results genotyped by PCR-SSP method, No novel nucleotide mutation was found at the exon 6 and exon 7 of ABO gene. In conclusion, the ABO genotyping assay by PCR-SSP provide a simple, rapid and accurate method for determining the ABO type of para-Bombay cases. PMID- 15363144 TI - [Clinical characteristics in 18 cases of sever pneumonia after allogeneic hematopoietic stem cell transplantation for malignant hematological disease]. AB - To investigate the clinical features and outcome of severe pneumonia after allogeneic hematopoietic stem cell transplantation for malignant hematological disease, the clinical features of severe pneumonia, including symptoms, signs, blood-gas analysis and thoracic X-ray change were observed, the clinical therapeutical outcome is also evaluated. The results showed that after treatment with antibiotics or antibiotics + antiviral drugs or antibiotics + antiviral drugs + glucocorticosteroids, as well as mechanical ventilation, pulmonary changes in 17 patients with sever pneumonia completely disappeared, 1 case died. In conclusion, early diagnosis and proper treatment for complicated pneumonia after allogeneic hematopoietic stem cell transplantation can decrease the mortality. PMID- 15363145 TI - [Effects of preparation and quality control of cryopreserved platelets]. AB - The purpose of this study was to establish a set of techniques for cryopreservation of platelets with dimethylsulphoxide (DMSO) to insure high quality of cryopreserved platelet. The methods were as following: (1) DMSO was filtered in stead of being sterilized before infusion into the bag with platelets. (2) The whole blood was centrifuged immediately after blood collection and the attached tube was tied on the top of the bucket. (3) The related centrifugal force was 480 x g, the accelerating and braking grades of the centrifuge for acceleration and deceleration were 9 and 4 respectively. (4) The flow rate of platelet rich plasma (PRP) could not be too high, 80 - 100 ml PRP should be harvested at 1 minute or so. The infusion rate of DMSO into the PRP was 1 ml/min. After the infusion of DMSO, the PRP bag must be put into the -80 degrees C ultra low freezer at once to make the product to be freezed as soon as possible. The cryopreserved platelet should be thawed in the cycling warm water at the temperature of 38 - 40 degrees C. (5) After thawing of platelet, the platelet, red blood cell and white blood cell were counted, and the bacteria culturing, tests for HBsAg, anti-HCV, anti-HIV, TP and ALT were carried out. The results showed that altogether 14 800 units of cryopreserved platelets were prepared including 80 units collected with blood cell separator, of which quality control was accomplished in 300 units. The manually collected platelet mean count >/= 2.4 x 10(10)/unit, while the apheresis platelet count >/= 2.5 x 10(11)/unit. The yield was over 70%. The contaminated red and white blood cells were papillary fibroblast-cultured = adipose fibroblast-seeded > papillary fibroblast-seeded > acellular carrier. After 21 days, only the acellular carriers were not vascularized and the papillary fibroblast-seeded constructs were not completely vascularized. Complete wound reepithelialization (92 +/- 12%) was observed only in the group treated with adipose cultured constructs. Wound contraction was not observed. Staining for HLA-ABC and alpha-smooth muscle actin showed that human fibroblasts had survived and that adipose fibroblasts continued to express the actin isoform. These results showed not only stimulation of skin repair when fibroblasts were present in the carrier, but also significant positive effects of the deposited extracellular matrix (ECM) in the carrier. In addition, the adipose fibroblast-seeded construct, and especially the adipose fibroblast-cultured construct, significantly stimulated angiogenesis and reepithelialization when compared with their corresponding papillary fibroblast constructs. Apparently, tissue source or fibroblast phenotype and the presence of ECM play a crucial role in the stimulation of (impaired) healing and engineering of dermal equivalents. PMID- 15363164 TI - Development of cell-seeded patellar tendon allografts for anterior cruciate ligament reconstruction. AB - Patellar tendon (PT) allografts for anterior cruciate ligament (ACL) reconstruction are potentially immunogenic and incorporate slowly compared with autografts. Our tissue-engineering approach to improve allograft efficacy is to (1) remove intrinsic cells from the graft to reduce antigenicity and then (2) seed the graft with extrinsic cells to improve ligamentization. To remove cells, tendons were soaked in 1% extraction solutions of tri(n-butyl)phosphate (TBP) or sodium dodecyl sulfate (SDS) for various time periods (24-72 h) and rinsed exhaustively. After treatment, we measured tendon cellularity, crimp structure, and mechanical properties. Treatment with either SDS or TBP removed approximately 70-90% of the intrinsic PT cells. Mechanical properties of treated PTs were similar to those of controls, despite changes in appearance. TBP- and SDS-treated PTs were then seeded with fibroblasts and cultured for up to 2 weeks in vitro. Fibroblast proliferation was retarded on SDS-treated PTs; in contrast, TBP treated PTs supported cell proliferation similar to that of untreated controls. Extrinsic fibroblasts were successfully cultured on the TBP-treated PTs in vitro, creating viable tissue-engineered grafts potentially useful for ACL reconstruction. These modified allografts have the potential to be developed into mechanically functional delivery vehicles for cells, gene therapy vectors, or other biological agents. PMID- 15363165 TI - Halofuginone inhibits serum-stimulated pericardial tissue retraction in vitro. AB - We developed an in vitro model of tissue contraction in which living pericardium, in response to serum, contracted and the cells in situ expressed proliferating cell nuclear antigen (PCNA) and synthesized collagen. Here we evaluated the effects of halofuginone on these serum-stimulated pericardial tissue responses. Parietal pericardium was incubated with media containing increasing doses of halofuginone and evaluated for tissue contraction, evident by tissue curling. Proliferation was measured by MTS metabolism and PCNA expression. Furthermore, collagen synthesis was compared between samples incubated with halofuginone, cytochalasin B, cytochalasin D, aphidicolin, or cytosine arabinoside (AraC), using Masson's trichrome and the monoclonal antibody to sheep type I procollagen, SP1. D8. Halofuginone inhibited tissue contraction, cellular proliferation, and collagen synthesis in a dose-dependent manner. In contrast, cytochalasin B, cytochalasin D, aphidicolin, and AraC, shown previously to prevent cellular proliferation, did not prevent type I collagen synthesis. Halofuginone has been implicated as an agent in the prevention of wound-healing fibrosis. This study suggests that halofuginone may have an added benefit in the inhibition of pericardial tissue contraction, which appeared to be related to the synthesis of type I procollagen. PMID- 15363166 TI - Composite articular cartilage engineered on a chondrocyte-seeded aliphatic polyurethane sponge. AB - To circumvent the reconstructive disadvantages inherent in resorbable polyglycolic acid (PGA)/polylactic acid (PLA) used in cartilage engineering, a nonresorbable, and nonreactive polyurethane sponge (Tecoflex sponge, TS) was studied as both a cell delivery device and as an internal support scaffolding. The in vitro viability and proliferation of porcine articular chondrocytes (PACs) in TS, and the in vivo generation of new articular cartilage and long-term resorption, were examined. The initial cell attachment rate was 40%, and cell density increased more than 5-fold after 12 days of culture in vitro. PAC-loaded TS blocks were implanted into nude mice, became opalescent, and resembled native cartilage at weeks 12 and 24 postimplantation. The mass and volume of newly formed cartilage were not significantly different at week 24 from samples harvested at week 6 or week 12. Safranin O-fast green staining revealed that the specimens from cell-loaded TS groups at week 12 and week 24 consisted of mature cartilage. Collagen typing revealed that type II collagen was present in all groups of tissue-engineered cartilage. In conclusion, the implantation of PAC-TS resulted in composite tissue-engineered articular cartilage with TS as an internal support. Long-term observation (24 weeks) of mass and volume showed no evidence of resorption. PMID- 15363167 TI - Allogeneic bone marrow-derived mesenchymal stromal cells promote the regeneration of injured skeletal muscle without differentiation into myofibers. AB - Half-stratum laceration was performed on the tibialis anterior muscle of Sprague Dawley (SD) rats as a skeletal muscle injury model. Bone marrow-derived mesenchymal stromal cells (BMMSCs), which were derived from enhanced green fluorescent protein (GFP) transgenic SD rats, were transplanted into the injured site. Tensile strength produced by nerve stimulation was measured for functional evaluation before sacrifice. Specimens of the tibialis anterior muscles were stained with hematoxylin and eosin, and immunohistochemically stained for histological evaluation. Our results showed that transplanted BMMSCs promoted maturation of myofibers histologically and made the injured muscle acquire almost normal muscle power functionally by 1 month after transplantation. However, the results of immunohistochemical staining could not prove that transplanted BMMSCs differentiated into or fused to skeletal myofibers, although it showed that transplanted BMMSCs seemed to differentiate into muscle precursor cells. Therefore, our results indicated that BMMSCs contributed to the regeneration of skeletal muscle by mechanisms other than fusion to myofibers after differentiation. PMID- 15363168 TI - Time course of primary liver cell reorganization in three-dimensional high density bioreactors for extracorporeal liver support: an immunohistochemical and ultrastructural study. AB - To enable extracorporeal liver support based on the use of primary liver cells, culture models supporting the maintenance of cell integrity and function in vitro are required. In this study the cell organization and ultrastructure of primary porcine hepatocytes cocultured with nonparenchymal cells in three-dimensional high-density bioreactors were analyzed after 10, 20, and 30 days of culture by immunohistochemistry and transmission electron microscopy. Biochemical data showed that metabolic activity of the cells in the system was relatively stable over at least 20 days. Immunohistochemical studies were performed in comparison with donor organ biopsies. They showed that hepatocytes and nonparenchymal cells reaggregated in bioreactors, forming structures partly resembling natural liver parenchyma. Bile duct-like structures characterized by cytokeratin 7 (CK-7) immunoreactivity (IR) were regularly detected. Nonparenchymal cells (vimentin IR) formed sinusoidal-like structures within parenchymal cell aggregates. Proliferative activity (Ki-67 IR) increased over time. The detection of collagen I and laminin indicated the production of extracellular matrix components within bioreactors. The results showed that primary liver cell reorganization and long term maintenance of their differentiated state were achieved within the bioreactors The findings on cell proliferation indicated that the culture model is also of interest for further in vitro studies on cell regeneration and tissue formation. PMID- 15363169 TI - Immobilization of cell-adhesive peptides to temperature-responsive surfaces facilitates both serum-free cell adhesion and noninvasive cell harvest. AB - We have developed temperature-responsive cell culture surfaces to harvest intact cell sheets for tissue-engineering applications. Both cost and safety issues (e.g., prions, bovine spongiform encephalopathy) are compelling reasons to avoid use of animal-derived materials, including serum, in such culture. In the present study, synthetic cell-adhesive peptides are immobilized onto temperature responsive polymer-grafted surfaces, and cell adhesion and detachment under serum free conditions were examined. The temperature-responsive polymer poly(N isopropylacrylamide) (PI-PAAm) was functionalized by copolymerization with a reactive comonomer having both a carboxyl group and an isopropylacrylamide group. These copolymers were covalently grafted onto tissue culture-grade polystyrene dishes. Synthetic cell-adhesive peptides were then immobilized onto these surfaces via carboxyl groups. Bovine aortic endothelial cells both adhered and spread on these surfaces even under serum-free conditions at 37 degrees C, similar to those in 10% serum-supplemented culture. Spread cells promptly detached from the surfaces on lowering culture temperatures below the lower critical solution temperature of the polymer, 32 degrees C. These surfaces would be useful for serumfree culture for tissue-engineering applications. PMID- 15363170 TI - Mesengenic progenitor cells derived from human placenta. AB - Progenitor cells with differentiation capacity along multiple mesengenic lineages are attractive tools for numerous purposes in regenerative medicine. Such mesengenic progenitor cells have been isolated from adult mammalian bone marrow, and we here report placental tissue as an alternative source for these cells. By means of dissection/proteinase digestion techniques, high numbers of viable mononuclear cells were harvested from human placenta at term, and a mesenchymal cell population with characteristic expression of CD9, CD29, and CD73 was obtained in culture. The in vitro growth behavior of such placenta-derived mesengenic cells was similar to that of human bone marrow mesengenic progenitor cells. After in vitro propagation for more than three passages the cells were exclusively of maternal origin. Differentiation experiments showed differentiation potential along osteogenic, chondrogenic, adipogenic, and myogenic lineages. In conclusion, we propose human term placenta as an easily accessible, ample source of multipotent mesengenic progenitor cells. PMID- 15363171 TI - Biomimetic collagen scaffolds for human bone cell growth and differentiation. AB - Type I collagen provides a structural framework for connective tissues and plays a central role in the temporal cascade of events leading to the formation of new bone from progenitors. The aim of this study was to examine the ability of the cell-binding domain of type I collagen (P-15 peptide) to promote human bone marrow stromal cell adhesion, proliferation, and differentiation on three dimensional scaffolds. Human bone marrow stromal cells were selected, expanded, and cultured on particulate microporous ABM ("pure" hydroxyapatite) phase adsorbed with or without P-15 under basal or osteogenic conditions. Immobilized P 15 increased alkaline phosphatase activity and bone morphogenetic protein 2 (BMP 2) gene expression after 1 and 5 days as determined by real-time polymerase chain reaction. P-15 promoted human bone marrow stromal cell attachment, spreading, and alignment on ABM as well as alkaline phosphatase-specific activity in basal and osteogenic cultures. The presence of mineralized bone matrix, extensive cell ingrowth, and cellular bridging between three-dimensional matrices adsorbed with P-15 was confirmed by confocal microscopy, scanning electron microscopy, and alizarin red staining. Negligible cell growth was observed on ABM alone. In vivo diffusion chamber studies using MF1-nu/nu mice showed bone matrix formation and organized collagen formation after 6 weeks. These studies indicate the potential of P-15 to generate appropriate biomimetic microenvironments for osteoblasts and demonstrate the potential for the exploitation of extracellular matrix cues for osteogenesis and, ultimately, bone regeneration. PMID- 15363172 TI - Electrospun nanofiber fabrication as synthetic extracellular matrix and its potential for vascular tissue engineering. AB - Substantial effort is being invested by the bioengineering community to develop biodegradable polymer scaffolds suitable for tissue-engineering applications. An ideal scaffold should mimic the structural and purposeful profile of materials found in the natural extracellular matrix (ECM) architecture. To accomplish this goal, poly (L-lactide-co-epsilon-caprolactone) [P(LLA-CL)] (75:25) copolymer with a novel architecture produced by an electrospinning process has been developed for tissue-engineering applications. The diameter of this electrospun P(LLA-CL) fiber ranges from 400 to 800 nm, which mimicks the nanoscale dimension of native ECM. The mechanical properties of this structure are comparable to those of human coronary artery. To evaluate the feasibility of using this nanofibrous scaffold as a synthetic extracellular matrix for culturing human smooth muscle cells and endothelial cells, these two types of cells were seeded on the scaffold for 7 days. The data from scanning electron microscopy, immunohistochemical examination, laser scanning confocal microscopy, and a cell proliferation assay suggested that this electrospun nanofibrous scaffold is capable of supporting cell attachment and proliferation. Smooth muscle cells and endothelial cells seeded on this scaffold tend to maintain their phenotypic shape. They were also found to integrate with the nanofibers to form a three-dimensional cellular network. These results indicate a favorable interaction between this synthetic nanofibrous scaffold with the two types of cells and suggest its potential application in tissue engineering a blood vessel substitute. PMID- 15363173 TI - Human mesenchymal progenitor cell-based tissue engineering of a single-unit osteochondral construct. AB - A desirable strategy for articular cartilage repair is to surgically replace the damaged area with an in vitro-engineered osteochondral plug. We report here the development of a novel osteochondral construct using human trabecular bone derived mesenchymal progenitor cells and a biodegradable poly-D,L-lactic acid scaffold. The cartilage layer was fabricated by press-coating a chondrifying high density cell pellet onto the scaffold, which was then loaded with cells previously initiated to undergo osteogenesis. The composite was then cultured in a cocktail medium formulated to maintain both chondrogenesis and osteogenesis. Macroscopically, the construct consisted of a cartilage-like layer adherent to, and overlying, a dense bone-like component. RT-PCR, immunohistochemistry, and histology revealed hyaline-like cartilage and bone with an interface resembling the native osteochondral junction. All parameters, including mechanical properties, improved with increased culture time. The single-cell source nature of the construct, which minimizes handling while maximizing biocompatibility, suggests applicability for articular cartilage repair. PMID- 15363174 TI - Survival of Apligraf in acute human wounds. AB - Apligraf consists of bovine collagen dermis seeded with allogeneic male fibroblasts and keratinocytes. It is been shown to promote healing, but the length of persistence and pathological features have not been characterized previously in acute wounds. Forty-eight deep dermal wounds were created and Apligraf, a split-skin graft (SSG), or a dressing was applied. Biopsies of wounds were taken for immunohistochemical analysis and polymerase chain reaction was performed to detect the Y chromosome from Apligraf cells in 14 female wounds. Male allogeneic DNA was detected in wounds for the first 4 weeks. All subsequent time points were negative apart from one biopsy at 6 weeks. The wounds took 4-9 weeks to heal, with the Apligraf exhibiting no features of engraftment. This was in contrast to the rapid healing seen in the SSG control group. Histology revealed a more intense cellular infiltrate, but less vascularization below Apligraf compared with controls. Evidence of an epidermal-mesenchymal interaction was observed. This is the first article to elucidate the survival of Apligraf allogeneic cells in acute wounds in immunocompetent human subjects for up to 6 weeks and demonstrates that in the management of acute surgical wounds, Apligraf has a role only as a temporary biological dressing. PMID- 15363175 TI - Controlled respiratory gas delivery to embryonic renal epithelial explants in perfusion culture. AB - During generation of artificial tissues high levels of oxygen are usually available whereas after implantation into a recipient's body the implant is not vascularized immediately, which leads to low oxygen partial pressures within the implanted tissue. Under these conditions cells will experience an oxygen shortage, contrasting with the abundance of oxygen during culture. It is uncertain whether tissues can be trained to tolerate such an acute hypoxic situation so that nonphysiological stress reactions and tissue necrosis can be avoided. To investigate the effects of varying oxygen levels on embryonic renal tissue in vitro we have been developing a model system combining continuous medium renewal with the ability to control levels of oxygen and carbon dioxide by gas equilibration through gas-permeable tubing. Renal embryonic tissue from neonatal rabbit was cultured in serum-free Iscove's modified Dulbecco's medium at 45, 90, 115, and 160 mmHg oxygen partial pressure for 14 days under continuous medium exchange in such a setup. After a 14-day culture period tissue sections were analyzed by cell biological methods and compared with fresh tissue histology. Surprisingly, embryonic renal explants survive and maintain good morphology for 14 days under all O(2) conditions tested. Expression of cytokeratin 19 within the established epithelium remains unchanged, indicating a structurally intact tissue. However, Na/K-ATPase is clearly downregulated under low O(2) conditions, whereas COX-2 expression increases drastically. An antiparallel effect of decreased O(2) concentrations on glycoprotein expression can be demonstrated with the lectin Dolichos biflorus agglutinin. Scanning electron microscopy reveals oxygen-dependent changes in cellular surface differentiation of developed collecting duct epithelium. PMID- 15363176 TI - Characterization of the cellular origin of a tissue-engineered human phalanx model by in situ hybridization. AB - Tissue-engineered models of human phalanges have previously been fabricated from a combination of bovine periosteum, cartilage, tendon, and biodegradable polyglycolic acid and poly-L-lactic acid scaffolds. Resulting constructs implanted in athymic mice for more than 40 weeks developed new bone, cartilage, and tendon and became vascularized, but cell types comprising the constructs were unidentified. The origin of cells in middle phalanx models implanted for 20 weeks in nude mice has been studied by in situ hybridization analyzing species-specific gene expression. Oligonucleotide probes homologous to species-specific gene sequences of bovine type II and X collagen, aggrecan, bone sialoprotein, biglycan, and osteopontin, and mouse decorin were labeled with (35)S and hybridized to respective serial sections of bovine tissue, mouse tissue, and phalanx constructs. In situ hybridization showed positive message and tissue specific localization for all bovine-specific probes examined within cartilaginous and midshaft portions of constructs and negative message for the mouse-specific decorin probe. These data show that osteoblasts and chondrocytes comprising constructs are derived exclusively from their original bovine sources over 20 weeks of implantation. Defining the cellular origin of the models lends insight into their biological, chemical, and physical nature and their growth and development. Maintenance of their initial genotype is crucial for future application of the models in augmenting impaired human phalanges and related tissues. PMID- 15363177 TI - Expression of alpha-smooth muscle actin by and contraction of cells derived from synovium. AB - Cells derived from synovium have drawn interest as donor cells for articular cartilage tissue engineering because they have been implicated in certain cartilage repair processes in vivo and the chondrogenic potential of the cells has been demonstrated in vitro. Studies have demonstrated that several other types of musculoskeletal connective tissue cells--including chondrocytes, fibrochondrocytes, ligament fibroblasts and osteoblasts, and mesenchymal stem cells can express the gene for the contractile actin isoform, alpha-smooth muscle actin (SMA), and can contract analogs of extracellular matrix in vitro. Although the physiological roles of SMA-enabled contraction of these cells have yet to be established, cell-mediated contraction of scaffolds employed for tissue engineering can alter the pore diameter of the matrix and distort its overall shape, and thus needs to be addressed. Toward this goal, the objective of this study was to investigate the expression of SMA by synovial cells and to evaluate their contraction of collagen-glycosaminoglycan (GAG) scaffolds. Synovial membranes obtained from the knees (stifle joints) of six adult dogs were evaluated for the presence of SMA by immunohistochemistry. Cells isolated from the synovial tissue were expanded through seven passages in monolayer culture, with samples from each passage allocated for Western blot analysis of SMA. Cells from passage 4 were seeded into porous type I collagen-GAG matrices and cultured for 4 weeks. Synovial cell-mediated contraction of the scaffolds was determined by measuring the diameters of the cell-seeded scaffolds and nonseeded controls every other day. Synovium-derived cells cultured as micropellets or in collagen GAG matrices were incubated in chondrogenic medium with and without fetal bovine serum and evaluated for chondrogenesis by type II collagen immunohistochemistry. Immunohistochemistry revealed the presence of SMA in some cells (less than 10% of the cells) in the intimal layer of synovium from four of the five animals analyzed. Western blot analysis demonstrated a regular increase in the amount of SMA in the synovium-derived cells with passage number. Synovial cell-mediated contraction of the collagen-GAG scaffolds reached a value of 43% of the original diameter after 4 weeks, comparable to that found with other musculoskeletal cell types. Incubation of micropellet cultures of synovium-derived cells with chondrogenic medium revealed trace amounts of type II collagen production by immunohistochemistry. The findings of this study indicate that control of SMA enabled contraction may be important when employing synovial cells for cartilage repair procedures, and warrant further investigation into the physiological role of SMA expression in synovial cells. PMID- 15363178 TI - Novel pulse duplicating bioreactor system for tissue-engineered vascular construct. AB - Cell culture in a biomimetic environment is known to improve the mechanical endurance of tissue-engineered cardiovascular components. Our goal was to generate a bioreactor that can reproduce a wide range of pulsatile flows with a completely physiological pressure profile. The morphology and biochemical properties of tissue-engineered products were also studied to test the usefulness of this novel bioreactor. The combination of an outflow valve, compliance chamber, and resistant clamps together with a balloon pumping system was able to successfully reproduce both physiological systolic and diastolic pressures. The compliance chamber was especially effective in transforming the original peaky pressure waveform into a physiological pressure profile. The tissues, cultured under a physiological pressure waveform with pulsatile flow, presented widely distributed cells in close contact with each other. They also showed significantly higher cell numbers, total protein content, and proteoglycan glycosaminoglycan content than cultured tissues under a peaky pressure wave or under static conditions. This new bioreactor system is suitable for evaluating a favorable environment for tissue-engineered cardiovascular components. PMID- 15363179 TI - In vivo tissue engineering of bone using poly(3-hydroxybutyric acid-co-3 hydroxyvaleric acid) and collagen scaffolds. AB - Porous poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) and calcium phosphate-loaded collagen (CaP-Gelfix) foams were seeded with rat bone marrow stromal cells and implanted into defects created in rat femurs to study in vivo bone formation and to test their suitability for use in bone tissue engineering. At 3 and 6 weeks, new bone formation was evaluated by macroscopy, radiography, dual-energy X-ray absorptiometry (DEXA), and quantitative computerized tomography (QCT). Atomic contents of the implants were further assessed by QCT. Some initial inflammation that significantly decreased with time was observed in the CaP Gelfix group. PHBV inflammation was minimal at all stages. Fibrous tissue formation in the CaP-Gelfix group was more than in the PHBV group. Both cell loaded and cell-free PHBV matrices elicited minimal fibrous tissue formation during the 6-week implantation duration. Macroscopic and radiological studies demonstrated better healing with PHBV matrices than with CaP-Gelfix in 3 weeks. Histologically, fibrous connective tissue establishment and inflammation scores were significantly higher in the CaP-Gelfix group when compared with the PHBV group at both time intervals. At 6 weeks, however, the extent of healing was almost the same with both implants. DEXA and QCT results indicated that there was an increase in bone mineral density in both PHBV and CaP-Gelfix implants at the end of 6 weeks. This study suggests that even though PHBV and CaP-Gelfix have different bulk and surface chemistries they both are promising cell carriers that may be suitable for use in bone tissue engineering. PMID- 15363180 TI - Regeneration in the ears of immunodeficient mice: identification and lineage analysis of mesenchymal stem cells. AB - Wound healing in the ears of Athymic Nude-nu mice resembles regeneration. Histological analysis of the ear-punched tissues revealed the initial formation of a blastema-like structure followed by dermal, vascular, cartilage, and muscle regrowth exclusively in Athymic Nude-nu mice but not in wild-type controls (C57BL/6J). A subset of stem cells referred to here as ear mesenchymal stem cells (EMSCs) has been isolated from the external ears of regenerative (Athymic Nude nu) and nonregenerative strains of mice. Morphological, histochemical, and molecular analysis after the induction of EMSC differentiation revealed multiple mesenchymal cell lineages (adipocytes, chondrocytes, and osteocytes) in all murine strains independent of their ability for regeneration. Thus, the absence of regeneration in wounded ears of C57BL/6J wild-type mice is not related to the absence of mesenchymal stem cell differentiation in tissue culture. Because nude mice lack T lymphocytes, it appears that in this model the absence of T lymphocytes in the wounded ears provides a microenvironment conducive to regeneration of mesenchymal tissues. These findings provide a new model to study the influence of the immune system on tissue regeneration. PMID- 15363181 TI - Spatial composition of prostate cancer spheroids in mixed and static cultures. AB - Aggregation of neoplastic cells produces multicellular spheroids resembling micrometastases. The objective of this study was to investigate the effects of mixing culture medium on the spatial composition of spheroids prepared from well (LNCaP) and poorly (DU 145) differentiated human prostate cancer cells. Spheroids were cultured in a mixed suspension within a high-aspect rotating wall vessel and static liquid-overlay plate. Results from this study demonstrate that mixed cultures consistently manifested differences in morphology and composition between DU 145 and LNCaP spheroids. For example, 40 +/- 12% of DU 145 cells were Ki-67 positive 100 microm from the surface within mixed spheroids versus 0% for LNCaP cells; there was no significant difference in this spatial profile for static cultures. The results suggest that poorly differentiated spheroids may be more likely to experience a change in composition from mixing culture medium than well-differentiated spheroids, due to low tissue density. Immunostaining for P glycoprotein is representative of this trend; average staining intensity increased 50% for DU 145 spheroids on mixing but was unchanged for LNCaP spheroids. The effects of mixing on spheroid composition were attributed to faster interstitial mass transport. Applications include drug development and delivery, as well as basic research on drug action and resistance. PMID- 15363182 TI - Tissue engineering of articular cartilage under the influence of collagen I/III membranes and low oxygen tension. AB - The objective of this study was to study the matrix production and phenotype stability of articular chondrocytes cultured on collagen I/III membranes (CM) under the influence of low oxygen tension (Po(2)). Primary bovine and osteoarthritic human chondrocytes were cultured for 2 weeks under 5-21% Po(2) on CM, in alginate, or as monolayers. Dedifferentiated cells were produced by 2-week monolayer culture under 21% Po(2). Collagen (Coll) type II and I expression was demonstrated immunohistochemically, by Western blotting (Coll II), and by semiquantitative RT-PCR; proteoglycan synthesis was demonstrated histochemically (toluidine blue); and biosynthetic activity was indicated by radiolabel incorporation ([(3)H]proline and [(35)S]sulfate). Bovine chondrocytes on CM showed an increase in Coll II expression and proteoglycan synthesis under low Po(2) conditions, whereas Coll I decreased. This oxygen-dependent phenotype stabilizing effect was even more pronounced in alginate cultures. Biosynthesis of bovine and human chondrocytes was also increased by low Po(2), except for proline incorporation, which decreased in bovine CM cultures (low-oxygen effects were significantly higher in alginate than in CM cultures). Dedifferentiated chondrocytes reexpressed Coll II protein when cultured under low Po(2) on CM or in alginate only, but not under high Po(2) or in monolayer culture. We conclude that CM and, even more, alginate foster phenotype stability and cartilage specific matrix production of bovine chondrocytes, especially when cultured under in vivo-like oxygen conditions. PMID- 15363183 TI - Effects of the degradation rate of collagen matrices on articular chondrocyte proliferation and biosynthesis in vitro. AB - The objective of this study was to evaluate effects of the degradation rate of type II collagen scaffolds on the proliferation and biosynthetic activity of adult canine chondrocytes in vitro. The lower number of cells in more rapidly degrading scaffolds appeared to be related to the loss of scaffold material with dissolution. After 14 days in culture, protein and proteoglycan synthesis rates per cell for rapidly degrading scaffolds were comparable to rates for nondegraded matrices. This result suggests that decoupling of the degradation and formation phases of tissue remodeling may occur under certain circumstances. PMID- 15363186 TI - [Prognosis and risk factors for recurrence of small liver cancer after a single session of percutaneous radiofrequency ablation]. AB - BACKGROUND & OBJECTIVE: As a new technique of local therapy for liver cancer, radiofrequency ablation (RFA) was widely used these years in China. This study was to evaluate the treatment efficacy of RFA on primary liver cancer (PLC), identify the risk factors of recurrence, and determine the indications of RFA for PLC. METHODS: Records of 94 primary small liver cancer patients underwent a single session of percutaneous RFA in Liver Cancer Institute/Zhongshan Hospital from Jan. 2001 to Dec. 2003 were reviewed retrospectively. Data analyses were performed using SPSS for windows Ver. 11.5 software. RESULTS: With a median follow-up of 16 months affer RFA treatment, the cumulative survival rate of 94 patients was 85.5% at 1 year, and 75.6% at 2 years. The cumulative recurrence free survival rate was 31.3% at 1 year, and 10.4% at 2 years. The total recurrence rate was 66.0% (62/94). Log-rank test revealed that tumor with diameter of >3 cm (P< 0.05), proximity to intrahepatic vessels (P < 0.01), and subcapsular location (P< 0.05) were related to tumor recurrence,while gender, Child-pugh class, alpha fetoprotein (AFP) concentration, and combination therapy with percutaneous ethanol injection were not related to tumor recurrence. Cox regression analysis indicated that tumor proximity to intrahepatic vessels (95%CI, 2.102-7.899; P=0.000), and subcapsular location (95%CI, 1.672-6.289; P=0.001)were associated independently with recurrence after RFA treatment. Severe complications occurred in 2 cases (2.1%), including 1 case of bile duct hemorrhage, 1 case of sub-diaphragm effusion. No RFA related death and other severe complications occurred. CONCLUSIONS: RFA appears to be a safe treatment for liver cancer. Patients with tumor diameter of 0.05). (2) The positive rate of P63 in EC was 81.6%, significantly higher than those in EH and BPE (P < 0.005). P63 expression was not associated with histological grade, surgical pathologic stage, and depth of myometrial invasion in EC (P >0.05). (3) The positive rate of C-erbB-2 in EC was 23.2%, there was no significant difference compared with those in EH or BPE (P >0.05).C-erbB-2 expression was associated with surgical pathologic stage, and depth of myometrial invasion in EC (P< 0.001,P< 0.005),but was not associated with histological grade (P >0.05).(4) There was significantly positive correlation between P53 and P63 (r =0.443,P < 0.01)or C-erbB-2 (r =0.490,P < 0.005). CONCLUSIONS: Both p53 and p63 are involved in carcinogenesis of EC; p63 may act as an oncogene in tumorigenesis of EC. The expression of P53 and C-erbB2 are related to the progression of malignant EC; P53 and C-erbB-2 co-expression may predict poor prognosis. PMID- 15363195 TI - [Human pituitary tumor transforming gene 1 expression in ovarian carcinoma and its clinical significance]. AB - BACKGROUND & OBJECTIVE: Human pituitary tumor transforming gene 1 (hPTTG1) is a newly identified oncogene. We performed a large-scale screening survey to identify related genes expressed in advanced and poorly differentiated serous ovarian carcinoma using cDNA microarray analysis, and found that hPTTG1 is one of highly up-regulated genes in ovarian carcinoma. This study was to examine hPTTG1 mRNA and protein expression in various epithelial ovarian carcinoma,analyze the relationship of its expression level with FIGO stage and histological grade, and explore the functional role of hPTTG1 in ovarian carcinoma pathogenesis. METHODS: HPTTG1 mRNA expression in 27 cases of epithelial human ovarian carcinoma, and 4 cases of normal ovary was assessed with non-competitive reverse transcription polymerase chain reaction (RT-PCR), and hPTTG1 protein expression in these 27 cases of human ovarian carcinoma, and 18 cases of normal ovary was examined by immunohistochemistry. RESULTS: HPTTG1 mRNA expression level in normal ovary was low, while in ovarian carcinoma was significantly higher than that in normal counterparts (P < 0.01). Fold increase of ovarian cancer tissues to normal ovary tissues is 1.1-4.8 (median 2.4).HPTTG1 mRNA high expression level was related to poor tumor differentiation (r = 0.686, P < 0.05), but the correlation with FIGO stage was not detected. HPTTG1 protein was expressed in all cases of ovarian carcinoma, but not in normal ovaries. CONCLUSIONS: Increased hPTTG1 expression may be an important factor involved in early tumorogenesis, and may be associated with poor tumor differentiation. HPTTG1 could be a potential molecular marker of epithelial ovarian carcinoma. PMID- 15363196 TI - [Effects of hypoxia on expression of HIF-1alpha,P53, and cyclin D1 in human lung adenocarcinoma cell line A549]. AB - BACKGROUND & OBJECTIVE: Many studies showed that hypoxia inducible factor-1 alpha (HIF-1alpha)was an essential component for hypoxia- induced cell cycle arrest, but the definite mechanism and the degree of HIF-1alpha affecting cell cycle arrest were unknown yet. This study was to explore the probable mechanism of hypoxia-induced tumor cell cycle arrest. METHODS: Human lung adenocarcinoma cell line A549 were divided into 3 groups: 12-h hypoxia group, 24-h hypoxia group, and control group. The hypoxia groups were exposed to hypoxic conditions (37degrees C, 5% CO2, and 2.0% O2) for 12 h, and 24 h, respectively, while control group was exposed to normal oxygen conditions (37 degrees C, 5% CO2, and 21% O2) for 24 h. Flow cytometry was used to measure the distribution of cell cycles and the expression of cyclin D1. The expression of HIF-1alpha, and p53 was detected using immunohistochemistry. RESULTS: (1) The ratio of G(0)/G(1) in 12-h hypoxia group was (70.20+/-3.33)%, and in 24-h hypoxia group was (82.85+/-1.75)%, significantly higher than that in control group [(50.36+/-4.09) %] (F=202.34, P< 0.01).(2) There was significant difference in cyclin D1 expression among 12-h hypoxia group [(80.22+/-1.55)%], 24-h hypoxia group [(73.65+/-2.10)%], and control group [(90.35+/-2.68)%] (F=100.45, P< 0.01). (3) HIF-1alpha expression in 12-hypoxia group, 24-hypoxia group, and control group was 0.16 +/- 0.02, 0.26 +/- 0.05, and 0.01 +/- 0.00, respectively, with significant difference among the 3 groups (F=105.28, P< 0.01). (4) In hypoxia groups, cyclin D1 expression was negatively correlated with G(0)/G(1) arrest (r=-0.91, P< 0.01), HIF-1alpha expression was positively correlated with p53 expression (r=0.84, P< 0.01), and negatively correlated with cyclin D1 expression (r=-0.90, P< 0.01), and p53 expression was negatively correlated with cyclin D1 expression (r=-0.78,P< 0.01). CONCLUSIONS: Hypoxia can cause G(0)/G(1) cell cycle arrest in human lung adenocarcinoma cell line A549. HIF-1alpha-p53-cyclin D1 pathway might play an important role in the hypoxia-induced G(0)/G(1) cell cycle arrest of human lung adenocarcinoma cell line A549. PMID- 15363197 TI - [Inhibitory effects of high mobility group box 1 antisense nucleotide on invasion of human pancreatic cancer cell line PCNA-1]. AB - BACKGROUND & OBJECTIVE: High mobility group box 1 (HMGB1) is abundantly expressed in most of immature, and malignant cells, and is closely related to cell proliferation, invasion, and migration. We have previously proved HMGB1 over expressed in pancreatic cancer tissues. This study was to evaluate the in vitro effects of HMGB1 antisense nucleotide on human pancreatic cancer cells invasion, and explore the underlying mechanism. METHODS: An eukaryotic expression vector containing antisense-HMGB1 was transfected into human pancreatic cancer cell line PCNA-1. The expression of HMGB1, matrix metalloproteinase-2 (MMP-2),and MMP-9 mRNA before and after transfection was detected by reverse transcriptase polymerase chain reaction(RT-PCR); HMGB1 protein expression was examined by Western blot. The activities of MMP-2, and MMP-9 were analyzed by gelatin zymography. The in vitro invasive ability of PCNA-1 cells was determined by Boyden chambers method. RESULTS: Antisense-HMGB1 eukaryotic expression vector was successfully constructed. HMGB1 mRNA expression in PCNA-1 cells was effectively inhibited by antisense-HMGB1. HMGB1 protein expression was also suppressed upon transfecting (P< 0.05). Antisense-HMGB1 transfected cells showed lower MMP-2 and MMP-9 mRNA expression (P< 0.01). MMP-2 and MMP-9 activities were also inhibited by antisens-HMGB1 (P< 0.01). Cells migrating through the Boyden chamber membrane was significantly reduced as compared with the control (P< 0.01). CONCLUSIONS: HMGB1 plays a crucial role in the invasion of pancreatic cancer, and blocking HMGB1 may be a potential strategy in preventing the migration of pancreatic cancer. PMID- 15363198 TI - [Quantitative detection of telomerase activity and its association with clinicopathological characteristics in breast cancer]. AB - BACKGROUND & OBJECTIVE: Telomerase is a ribonucleoprotein enzyme, which plays an important role in cell immortalization and carcinogenesis. Recent studies on the association of telomerase activity with prognostic factors of breast cancer were controversial due to different methods. This study was to establish a feasible assay of quantitative detection of telomerase activity based on silver staining, and investigate possible association between telomerase activity and clinicopathological prognostic factors in primary breast cancer. METHODS: Highly sensitive silver-staining telomeric repeat amplification protocol assay (SS-TRAP) was used to quantify telomerase activity in 52 frozen human breast cancer samples and their adjacent breast tissues, 32 benign lesions, and 14 normal mammary gland lesions. The association between telomerase activity and clinicopathological data was analyzed. RESULTS: Telomerase activity was detected in 47 of the 52 breast cancer samples (90.38%), and in 10 of the 32 benign lesions, whereas no activity was detected in 37 of 52 adjacent nonmalignant breast tissues, and all 14 normal mammary gland tissues. The telomerase activity levels were 36.91+/-15.35, 8.27+/ 4.37, 14.10+/-5.28, 0 (unit: TPG) in breast cancer, adjacent tissue of cancer, benign lesion, and normal tissue, respectively. The difference of telomerase activity was significant between breast cancer and the other 3 groups by using ANVOA (all P< 0.01). A strong correlation was found between telomerase activity and pathological category, and differentiation degree by logistic regression analysis, i.e. with ongoing tumor progression, telomerase activity appeared to increase in primary breast cancer (P=0.003, and P=0.004). No correlation was seen between telomerase activity and disease course, age, and menopause status of patients (all P >0.05). Telomerase activity level of invasive non-special cancer was higher than that of invasive special cancer(P< 0.05). Telomerase activity level of moderately/poorly differentiated carcer was higher than that of highly differentiated cancer (P< 0.05), while no obvious difference was found between moderately and poorly differentiated cancer (P >0.05). CONCLUSIONS: The activation of telomerase might occur early in breast cancer,and plays a critical role in carcinogenesis and tumor development. Telomerase may serve as a specific marker of early diagnosis and prognosis in mammary gland neoplasm. PMID- 15363199 TI - [Effects of transfected SOCS3 gene on proliferation of human lung adenocarcinoma cell line A549]. AB - BACKGROUND & OBJECTIVE: Previous studies showed that signal transducers and activators of transcription 3(STAT3) could be constantly activated in many kinds of tumor cells; constantly activated STAT3 could make cell malignant transformation, and enhance the malignant proliferation of tumor cells. The suppressors of cytokine signaling (SOCS) proteins, a recently identified family, could suppress the tyrosine-phosphorylated process of STAT protein to negatively regulate STAT-mediated cytokine signaling, which is closely related to cell survival, proliferation, and apoptosis. Among the SOCS protein family, SOCS3 is the most potent inhibitors of cytokine-induced signaling. This experiment was to investigate the effect of SOCS3 on proliferation of human lung adenocarcinoma cell line A549. METHODS: A549 cells were co-transfected with pEFSOCS3 and pSV2neo by liposome, expression of SOCS3 mRNA and protein was determined by reverse transcriptase-polymerase chain reaction (RT-PCR), and immunocytochemistry. Tyrosine-phosphorylated STAT3 level was measured by Western blot analysis. MTT assay and (3)-TdR incorporation were used to measure the cell proliferation in pEFSOCS3-transfected group, and control group (non-transfected, and pEF transfected A549). RESULTS: Compared with control group, tyrosine-phosphorylated STAT3 level suppressed greatly in pEFSOCS3-transfected A549 cells (P< 0.01). The inhibitory rate of cell growth in pEFSOCS3-transfected group was 40.58%, and in control group was 32.23%. (3)-TdR incorporation (48 056+/-1 331) in pEFSOCS3 transfected group was significantly lower than that of control group (81,481+/ 2,584, and 75,590+/-2,678, P< 0.01). CONCLUSION: SOCS3 protein might inhibit proliferation of A549 cells by lowering the tyrosine-phosphorylated level of STAT3 protein. PMID- 15363200 TI - [Relationship between the expression of O6-methylguanine-DNA methyltransferase in glioma and the survival time of patients]. AB - BACKGROUND & OBJECTIVE: Previous studies showed that the DNA-repair enzyme O(6) methylguanine-DNA methyltransferase (MGMT) is one of drug resistant factors that affect chemosensibility of glioma. This study was to analyze the relationship between the expression of MGMT in glioma and the survival time of patients,and supply references to make molecular classification for glioma based on drug resistant mechanism. METHODS: MGMT expression in 311 glioma specimens was examined by tissue array technology and immunohistochemistry method, all patients had been followed up for 5 years, and the materials were analyzed statistically. RESULTS: The positive expression of MGMT was 126 in 311 gliomas (40.51%), among them, 61 in 121 astrocytomas (50.41%), 18 in 70 oligodendrogliomas (25.71%), 18 in 64 oligoastrocytomas (28.13%), 29 in 56 glioblastomas (51.79%); 68 in 186 grade I-II gliomas (36.56%), and 58 in 125 grade III-IV gliomas (46.40%). The difference of MGMT expression between grade I-II and grade III-IV gliomas was significant (P< 0.001). According to Kaplan-Meier's survival curves and log-rank test, patients with MGMT expression showed a shorter survival time than those with no MGMT expression (P< 0.05). CONCLUSION: MGMT expression in glioma correlates with histopathological type and tumor grade. Patients with MGMT expression show a shorter survival time than those with no MGMT expression. PMID- 15363201 TI - [Analysis of computed tomography-based distribution of metastatic cervical nodes in 218 cases of nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: The application of intensity- modulated radiation therapy (IMRT) for nasopharyngeal carcinoma (NPC) requires a precise delineation of the nodal area and nodal clinical target volume (CTV) on computed tomography (CT) images,and the prerequisite is to find out the rules of CT-based distribution of metastatic lymph nodes of NPC. This study was designed to analyze the rules of CT based distribution of nodal involvements of NPC according to the guidelines of nodal levels proposed by Radiation Therapy Oncology Group (RTOG). METHODS: From Jul. 2003 to Nov.2003, 259 newly diagnosed NPC patients received radiotherapy at Fudan University Affiliated Cancer Hospital. All patients had transversal contrast enhanced CT scan from base of skull to clavicle before treatment. Diagnostic radiologists and radiation oncologists together assessed the nodal distribution in each RTOG nodal level. Chi-square test was used to analyze the correlation between T stage and nodal metastasis rate. The neck was further divided into 3 regions by the verge of hyoid bone and the inferior border of cricoid cartilage to assess leap metastasis of nodes. RESULTS: A total of 218 patients (84.2%)had nodal involvement. The distribution was as follow: 0 in level Ia, 6 (2.8%) in level Ib, 115 (52.8%) in level IIa,192 (88.1%) in level IIb, 78 (35.8%) in level III, 20 (9.2%) in level IV, 65 (29.9%) in level V, 0 in level VI,157 (72.0%)in retropharynx, and 2 (0.9%) at preauricular area. Leap metastases were found in only 5 patients (2.3%). No significant correlation was found between T stage and nodal involvement. CONCLUSION: NPC has a high probability of nodal metastases, nodes in level IIa,IIb, and retropharynx are most likely to be involved. Nodes metastasized mostly from the upper to the lower level, and from the proximal to the distal part, with a very low leap metastasis rate. The relationship between T stage and nodal involvement has no statistical significance. PMID- 15363202 TI - [Clinical significance of quantitative analysis of serum VEGF, CD44s, and MMP-3 protein in nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: Increasing evidences showed that circulating tumor metastasis related factors could be detected, and it is related to tumor stage, and prognosis. This study was to investigate the correlation of serum vascular endothelial growth factor (sVEGF), CD44 standard (CD44s), and matrix metalloproteinase-3 (MMP-3) level to the clinical outcomes in nasopharyngeal carcinoma (NPC)patients. METHODS: Forty-six NPC patients without metastasis, 20 NPC patients with local recurrence and/or distant metastasis,and 28 healthy controls entered this study. Serum VEGF, CD44s, and MMP-3 protein were quantitatively analyzed by enzyme-link immunosorbent assay (ELISA). RESULTS: The mean level of sVEGF in metastatic NPC was (791.7+/-560.5) ng/L, which was significantly higher than that in NPC patients without metastasis [(429.0+/ 249.7) ng/L], and healthy controls [(424.6+/-197.1) ng/L], whereas there was no significant difference between NPC patients without metastasis and healthy controls. In primary NPC patients, sVEGF level in NPC of stage T4 were significantly higher than those in NPC of stage T1-T3. Moreover, 2-year disease free survival rate in patients with sVEGF level of more than 600 ng/L was significantly lower than that in patients with sVEGF Level of less than 600 ng/L (37.5% vs 83.9%). The serum level of MMP-3 in NPC patients with metastasis [(28.8+/-15.5)microg/L] was significantly higher than that in patients without metastasis [(19.8+/-11.6) microg/L], and healthy controls [(16.2+/-11.1) microg/L], but there was no significant difference between NPC patients without metastasis and healthy controls. Though the serum level of CD44s in NPC patients (including patients with or without metastasis) was higher than that in healthy controls, there was no significant difference between NPC patients with and without metastasis. No correlation was observed between serum MMP-3, CD44s level and clinical outcomes of NPC. CONCLUSION: Our results indicate that when the sVEGF level are >or= 800 ng/L and/or serum MMP-3 level >or= 30 microg/L in NPC patients, local recurrence or distant metastasis would occur. In primary NPC patients without metastasis, sVEGF level is correlate with primary tumor progression, moreover, sVEGF level of >or= 600 ng/L before treatment predicate lower 2-year disease-free survival rate. PMID- 15363203 TI - [Correlation of the changes of T lymphocyte phenotype to tumor stage and operative pattern of gastric cancer]. AB - BACKGROUND & OBJECTIVE: Immunity, especially cellular immunity, of patients with tumor is related to tumorigenesis. The correlation of changes of T Lymphocyte phenotype to tumor stage and operative pattern of gastric cancer is unclear. This study was to evaluate the perioperative immune state in patients with gastric cancer (GC) of different stage and accepted different operative pattern. METHODS: Six kinds of T lymphocyte phenotype in 33 GC patients were measured by flow cytometry before and after operation, and compared with that of benign disease patients. RESULTS: With progress of cancer stage, CD3, CD4, CD4/CD8, CD16, and CD69 gradually decreased, while CD8 gradually increased (P< 0.05). There was no significant difference of activated T cell CD3+/HLA-DR+ among patients of stage I,II,III. After radical resection, CD8 decreased, while CD3, CD4, CD4/CD8, CD16, CD69, and CD3+/HLA-DR+ increased significantly (P< 0.01). CD3, and CD4 unchanged after palliative operation. CD16, and CD4/CD8 further decreased in patients with unresectable tumor (P< 0.05). CONCLUSIONS: The preoperative immune state of GC patients is negatively related to cancer stage. Tumor removal may improve the cellular immunity of patients. PMID- 15363204 TI - [Clinical significance of detecting lymph node micrometastasis of colorectal cancer by reverse transcriptase-polymerase chain reaction (RT-PCR)]. AB - BACKGROUND & OBJECTIVE: It remains controversial whether lymph node micrometastasis has impact on staging and prognosis of colorectal cancer. This study was to compare the sensitivity of reverse transcriptase- polymerase chain reaction (RT-PCR) in detecting lymph node micrometastasis of colorectal cancer with pathological morphology and immunohistochemistry, and assess the impact of lymph node micrometastasis on clinical staging and prognosis of colorectal cancer. METHODS: Lymph nodes from 56 cases of colorectal cancer radical resection specimens were studied by RT-PCR to detect the expression of cytokeratin 20 (CK20) mRNA, and compared with routine pathology detection using hematoxylin and eosine (HE) staining, and immunohistochemistry using monoclonal antibody specifically against CK20. The patients had been followed up for 5 years. RESULTS: A total of 432 lymph nodes in 56 patients were analysed by pathological morphology, immunohistochemistry, and RT-PCR, the detected positive lymph node numbers were 247 (57.2%), 269 (62.3%), and 316 (73.1%), respectively. The difference in metastatic lymph node numbers was significant between pathological morphology and RT-PCR method (P< 0.05). Five-year disease- free survival rates of PN0,PN1, and PN2 stages detected by RT-PCR method were 100%, 61.9%, and 55.6%, respectively, significantly higher than those obtained by pathological morphology method, which were 80.0%, 60.0%, and 50.0%, respectively (P< 0.05). CONCLUSIONS: Detecting lymph node micrometastasis of colorectal cancer with RT-PCR method is more sensitive than pathological morphology. RT-PCR method could define the TNM stage and make accurate prognosis for patients with colorectal cancer. PMID- 15363205 TI - [Diagnosis and treatment of renal cell carcinoma with vena cava tumor thrombi]. AB - BACKGROUND & OBJECTIVE: Renal cell carcinoma (RCC) might involve the renal vein, and form tumor thrombus extending into the vena cava or the right atrium. Treating RCC with vena cava involvement is difficult in clinical practice. Radical nephrectomy with complete tumor thrombus removal could result in good outcomes for RCC patients with vena cava thrombi. This paper was to report our experiences on treating RCC with vena cava thrombi. METHODS: From May 1995 to Oct. 2003, radical nephrectomy plus vena cava thrombus removal was performed in 14 RCC patients. Clinical records, including preoperative diagnosis, operation pattern, and prognosis, of these 14 patients were analyzed retrospectively. RESULTS: Vena cava thrombi were detected in 9 patients, and missed in 5 patients by ultrasonography. CT scan revealed vena cava thrombi in 12 of 14 patients. MRI has been performed in 8 patients, and clearly demonstrated extent of the thrombus. Twelve cases of tumor thrombi within infrahepatic vena cava, 1 within intrahepatic subphrenic vena cava, and 1 within supraphrenic vena cava. The patients have been followed up for 6-70 months after surgery. Thirteen patients survived with disease-free, and 1 patient (stage IIIc) died of cancer 23 months after surgery. CONCLUSION: Vena cava thrombi in patients with RCC could be detected on CT scan and ultrasonography. MRI is more accurate than CT,and ultrasonography in delineating extent of the thrombus. Radical nephrectomy plus vena cava thrombus removal could achieve long-term survival for patients with localized RCC and vena cava thrombi. PMID- 15363206 TI - [Meta analysis of the relationship between tumorigenesis of oral squamous cell carcinoma and human papillomavirus infection]. AB - BACKGROUND & OBJECTIVE: Previous researches showed that human papillomavirus (HPV) infection was closely related to tumorigenesis of oral squamous cell carcinoma (OSCC). But the results of these studies have great differences because of different research methods. This study was to evaluate synthetically the relationship between OSCC of Chinese and HPV infection by meta analysis. METHODS: From Jan. 1990 to Apr. 2003, 44 references reported about the relationship between tumorigenesis of OSCC of Chinese and HPV infection were collected from Chinese Biomedical Literature Analysis and Retrieval System for Compact Disc (CBMdisc). There were 10 references accorded with research criteria which was case-control study, and detected by polymerase chain reaction (PCR). Fisher and meta analysis were used to quantitatively and qualitatively analyze these references synthetically. RESULTS: The synthetic qualitative analysis by Fisher showed there was significant association between the tumorigenesis of OSCC and HPV infection (P< 0.005). The synthetic quantitative analysis by meta analysis revealed that the combined odds ratioes (ORc) for HPV, and HPV16 infection of OSCC were 8.89 (3.62-21.80), and 6.81 (2.18-21.32) times that of normal oral mucosa. The mean positive rates of HPV, and HPV16 detection in OSCC were 49.02% (36.48%-61.57%),and 45.74% (31.94%-59.54%) higher than that of normal oral mucosa. CONCLUSION: Human papillomavirus infection, especially HPV16, will increase the risk of tumorigenesis of OSCC. PMID- 15363207 TI - [Poorly differentiated thyroid follicular carcinoma - a clinical analysis of 30 cases]. AB - BACKGROUND & OBJECTIVE: Poorly differentiated thyroid follicular carcinoma is rare, and there were few reports on it. This study was designed to determine clinical characteristics, treatment, and prognosis of poorly differentiated thyroid follicular carcinoma. METHODS: Thirty patients with poorly differentiated thyroid follicular carcinoma were investigated retrospectively for their general clinical manifestation, process of treatment,and prognosis. Duration of survival from operation to various endpoints, including local recurrence, distant metastasis, and death from cancer or any causes, was estimated by Kaplan-Meier method. Factors that might be related to prognosis, including age, gender, tumor size, extrathyroidal extension, treatment, and operation status, were analyzed by log-rank test. The difference of data between 2 groups was estimated by t test. RESULTS: The 3-, 5-, 10-year overall survival rates of 30 patients with poorly differentiated thyroid follicular carcinoma were 32.2%, 25.1%, and 12.5%, respectively. The cumulative recurrence rate, regional lymph node metastasis rate, and distant metastasis rate were 37.6%, 57.6%, and 82.5% at 3 years, and 48.0%, 68.2%, and 86.9% at 5 years, respectively. The local recurrence rate decreased significantly (Chi(2)=6.59,P=0.01), and the survival rate increased significantly (Chi(2)=12.40,P=0.00) for patients who had complete removal of the primary lesion and neck metastasis. The recurrence rate of patients with venous tumorous emboli was remarkably higher than that of other patients,(Chi(2)=4.62,P=0.03). The survival time of patients with distant metastases, who had undergone 131I therapy, was significantly longer than that of other patients, who had not undergone 131I therapy (Chi(2)=12.25,P=0.00). CONCLUSION: Patients with poorly differentiated thyroid follicular carcinoma had high distant metastasis rate, and poor prognosis. Distant metastasis is an important factor that affects the survival rate. Radical resection is important for control primary tumor, and improve survival rate. 131I therapy is an effective treatment and could prolong the survival time of patients with distant metastases. PMID- 15363208 TI - [Multivariate analysis of prognostic factors in endometrial carcinoma]. AB - BACKGROUND & OBJECTIVE: A number of prognostic factors have been evaluated in endometrial carcinoma. Among them, only a few factors have been recognized as major adverse factors affecting prognosis independently. This study was to explore the independent prognostic factors in endometrial carcinoma. METHODS: A total of 265 patients who accepted primary surgical treatment in our hospital from Jan. 1990 to Dec. 2000 were studied retrospectively. Prognostic factors were analyzed using univariate model and multivariate regression model. RESULTS: The 5 year recurrence-free survival rate of all patients was 83.3%, and 5-year overall survival rate was 84.3%. The univariate model revealed that clinical stage, pathological stage, pathologic grade, pathologic subtype, depth of muscularis invasion, cervical invasion, lymph node metastasis, peritoneal cytology, lymph vascular invasion, and adnexa metastasis significantly associated with 5-year recurrence-free survival, and 5-year overall survival (P< 0.05), while age, and complications have no significant association with prognosis (P >0.05). However,on multivariate regression analysis, only pathohistological stage, pathologic grade, depth of muscularis invasion, and cervical invasion significantly correlated with 5-year recurrence-free survival, and 5-year overall survival (P< 0.05); clinical stage correlated with 5-year recurrence-free survival significantly (P< 0.001), but not with 5-year overall survival (P=0.074). Hematogenous dissemination rate of patients with muscularis invasion of >50% was significantly higher than that with muscularis invasion of 0.05) and grade of the tumor (chi2 = 0.45, 0.04, P > 0.05); but it correlated with the clinical stage (chi2 = 26.6, 20.1; P < 0.01) and survival time of the patients (chi2 = 21.5, 22.2; P < 0.01). CONCLUSIONS: The results suggest that there be overexpression of HPSE and VEGF-C protein in lung cancer tissues, and which perhaps participate in regulation of tumorigenesis, progression in lung cancer. The expressions of HPSE and VEGF-C protein are used as an useful marker of the biological behavior of lung cancer and as an independent prognosis factor for the patients with lung cancer. PMID- 15363234 TI - [Effect of T suppressor cells on the maintenance phase of tolerance to cardiac allografts in the rats]. AB - OBJECTIVE: To study the role of T suppressor cells in immune tolerance to cardiac allografts in the rats. METHODS: Male DA rat hearts were transplanted to male Lewis rats using Ono's model and randomly divided into five groups: group 1: untreated, group 2: portal venous injection of 3 x 10(8) DA splenocytes to Lewis rat, group 3: intraperitoneal injection of cyclophosphamide (80 mg/kg) to Lewis rat, group 4: portal venous injection of 3 x 10(8) DA splenocytes combined with intraperitoneal injection of cyclophosphamide (80 mg/kg) to Lewis rat, 15 days later heart transplantation was performed. Group 5: intravenous injection 3 (108 splenocytes of group 4 to normal recipient, and then heart transplantation was performed. Mean survival time (MST), histological changes, mixed lymphocyte reaction (MLR) were measured after operation. RESULTS: The survival time of heart allografts in the group 4 [MST: (71.5 +/- 29.1) d, t = -14.063, -13.915, -13.777; P < 0.01] was significantly longer than in the groups of 1 [MST: (7.3 +/- 1.0) d], 2 [MST: (7.8 +/- 0.8) d], 3 [MST: (8.2 +/- 1.1) d ]. Only a few inflammatory cells infiltrated in cardiac allografts in the groups of 4 and 5. MLR in the groups of 4 and 5 were significantly decreased compared with those of normal control (t = 29.902, 23.047; P < 0.01). CONCLUSIONS: Portal venous injection of donor splenocytes combined with intraperitoneal injection of cyclophosphamide could induce immune tolerance to cardiac allografts. The immune tolerance could be transferred through splenocytes. T suppressor cells play an important role in the immune tolerance. PMID- 15363235 TI - [Relationship between tunnel widening and different rehabilitation procedures after ACL reconstruction with quadrupled hamstring tendons]. AB - OBJECTIVE: To find out the relationship between different rehabilitation procedures and the tibial tunnel widening after ACL reconstruction. METHODS: Sixty-five patients undergone ACL reconstructions by using quadrupled semitendinosus and gracilis tendons were divided into two groups. Group A had 33 patients, 19 male, 14 female, average age (31.2 +/- 12.4) years, only ACL reconstruction was performed with Germanic Professor Paessler's technique, and aggressive rehabilitation procedure was used for functional recovery. Group B had 32 patients, 20 male, 12 female, average age (30.3 +/- 10.3) years. Besides ACL reconstruction, every patient in Group B accepted meniscus repair or cartilage repair with microfracture technique, then postoperative conservative rehabilitation procedure was used. Six months after operation, standard posterior anterior radiographic plates were taken for each patient, CorelDRAW 8.0 software was used to digitize all X-ray plates and measure the upper, middle and lower parts of the tibial tunnel. Magnification of X-ray plates was taken out after measurement. Statistic analysis was done by t-test. RESULTS: Six months after ACL reconstruction, on both the posterior-anterior and lateral X-ray plates the tibial tunnel widening of the upper, middle and lower parts in Group B with aggressive rehabilitation procedure was significantly serious than in Group A with conservative rehabilitation. CONCLUSIONS: Rehabilitation procedure after ACL reconstruction is one of the reasons for tunnel widening. It perhaps influences the functional recovery and long-term clinical result of the operated knee by affecting the tunnel widening. PMID- 15363236 TI - [A prospective study of the anterior cruciate ligament reconstruction: allograft versus autograft]. AB - OBJECTIVE: To compare the clinical outcomes and differences of arthroscopic anterior cruciate ligament (ACL) reconstruction by using allografts and autografts. METHODS: Fifty-three patients with grade III ACL injuries were divided randomly into 2 groups. Twenty-five cases of group A with autografts of patellar bone-tendon-bone (B-PT-B;n = 15) and 4 strands of semitendinosus/gracilis (n = 10); 28 patients of group B with allografts of B-PT B (n = 18), 4 strands of semitendinosus/2 strands of gracilis tendons (n = 6) and double tibialis posterior (n = 4) as well as Achilles bone-tendon (n = 2). All procedures were performed endoscopically by some surgeons. All patients were followed up for 12 to 31 months, average 19 months. RESULTS: There were statistically significant differences revealed in physical examination. IKDC and Lysholm-Tegner Score as well as KT-2000 testing was used preoperation and postoperation for both group, but there were no significant differences between the two groups except that group B had less time in operation and longer time in postoperative fever comparing with group A. (2-sample t test, P < 0.05). The KT 2000 side to side difference less than 3 mm were 88% and 86% and more than 5 mm were 4% and 7.1% respectively. The infection rates were 0% and 3.5% for the two groups. CONCLUSION: The clinical outcomes of the allografts and autografts in ACL reconstruction is almost the same. Allografts are a reasonably alternative choice for ACL reconstruction in patients over middle ages and for multiple ligament surgery as well as revision procedures. PMID- 15363237 TI - [Ilizarov technique for correcting flexion deformity of the knee of arthrogryposis multiplex congenita]. AB - OBJECTIVE: To study the methods and effects of Ilizarov distraction technique in treating the flexion deformity of the knee of arthrogryposis multiplex congenita. METHODS: Between August 1998 and February 2003, 6 patients (10 knees) with the arthrogryposis multiplex congenita were treated, 4 patients in double knees, 5 males, 1 female, mean age 8 years and 2 months, ranged from 3 years and 7 months to 13 years. The preoperative flexion degree was averagely 51 degrees. The patients accompanied 13 other parts malformation of limbs. The modified Ilizarov distraction apparatus of the knee was used. While installing the apparatus in the operation, the knees should be kept in the of location of maximum extension, the center of joint hinges on the apparatus should be placed towards the rotatory center of the knee, two groups of 2 mm K-wires were passed through the femur and tibia around the knee, who were fixed on the proximal and distal rings. Distraction was started after the surgical procedure 5 days via rotating the threaded rods at the posterior of the knee, at an average of 2 to 3 mm per day, at the first week, after 2 weeks with the rate modified to 2 mm per day, up to the knee extended to 0 degrees. The accompanied deformities of the hip and/or the foot might be corrected at the same time or next time. The average duration of the distraction was 37 days (23-48 days). During the correction all limbs might undergo weight. After 2 weeks at the end of distraction the fixator was removed and the patients could walk by a long-leg brace. RESULTS: Ten knees with the flexion contracture were sufficiently corrected without severe complications. Nine knees of all were followed up at an average time of 1 years and 3 mouths, no recurrence of the deformity was seen in all patients, their function of walk was significantly improved. CONCLUSIONS: Ilizarov technique is a simply, safe and effective method for managing the flexion deformity of the knee of the arthrogryposis multiplex congentia. The procedure is conformable to the biological theories and microsurgical principles. PMID- 15363238 TI - [Restoration of femoral offset in total hip arthroplasty]. AB - OBJECTIVE: To explore the factors effecting restoring femoral offset and the relation between femoral offset and hip abductor strength during total hip arthroplasty (THA). METHODS: Ninety-nine THA for 81 patients were performed from March 1998 to January 2002. And follow-up was finished. There were 53 women and 28 men and the average age was 57 years (29 to 80). The right hip had been replaced in 28 cases, the left in 35 cases and the bilateral in 18 cases. The mean duration of clinical and roentgenographic follow-up was 36.8 months (range, 19 approximately 66 months). A posterolateral approach was used in all THA. The femoral offset and the abductor lever arm were measured from each radiograph. The measurement of the hip abduction strength was made for some THA by the Cybex machine. Statistical data analysis was performed by SPSS10.0 software. RESULTS: Femoral offset correlated positively with the length of the abductor lever arm (r = 0.613; P < 0.001). Simple regression analysis showed that femoral offset was significantly and positively related to the length of femoral neck and neck-shaft angle (r = 0.451, P = 0.001; r = 0.567, P < 0.001). There was a highly significant and positive correlation between femoral offset (and consequently abductor lever arm) and hip abductor strength (r = 0.500, P = 0.009; r = 0.477, P = 0.014). CONCLUSIONS: It is very important to template both sides of hip preoperatively for restoring femoral offset in THA. Femoral component with more anatomical neck-shaft angle will be used with the increase in the femoral neck length. PMID- 15363239 TI - [Acetabular reconstruction of total hip replacement in the treatment of dysplastic hip]. AB - OBJECTIVE: To summarize the surgical experience of acetabular reconstruction of total hip replacement in the treatment of dysplastic hip. METHODS: From April 1998 to April 2002, 96 adult cases with osteoarthritis secondary to dysplastic hip were adopted in the study. Among them, there were 89 females and 7 males, age from 27 approximately 68 years. According to Hartofilakidis classification system, there were dysplasia in 73 hips, low dislocation in 18 hips and high dislocation in 21 hips. All patients received total hip replacement, 16 cases with bilateral replacement and 80 cases with unilateral replacement. Kocher LangenBeck approach was used during operation and all the acetabular cups were reconstructed at the original anatomic location. The fixation methods were as follows: cement cup for 16 hips, cementless cup for 96 hips and bone graft in 11 hips. The reconstructive methods were as follows: regular replacement for 83 hips; installing a smaller cup after deeper reaming the acetabulum for 27 hips; installing a smaller cup after autofemoral grafting on the superior lip for 2 hips. RESULTS: The incision healed primarily and no infection or nerve injury occurred. Follow-up for 1 approximately 5 years (average for 3.5 years) in 85 patients, the Harris score increased from 33.9 preoperatively to 89.3 postoperatively, and 95 hips had good or excellent clinical results. Radiographically, the positions of the prostheses were normal, the average abduction angle of the cup was 44 degrees, the average superolateral bone coverage of the hips was 96.6%, no radiolucent line was observed at the acetabular side. All the bone grafts fused with the host bone successfully. CONCLUSIONS: Total hip replacement is an effective operation in the treatment of osteoarthritis secondary to dysplastic hip. The four specific factors that should be considered in acetabular reconstruction are: (1) the surgical approach, (2) the reconstructive position of the acetabulum, (3) the reconstructive method of the acetabulum, (4) the leg-length discrepancies. It's a challenging to the orthopaedic surgeon. Careful preoperative preparation is recommended. PMID- 15363240 TI - [Total hip arthroplasty with uncemented cup and femoral head autografts for coxarthrosis due to dysplasia]. AB - OBJECTIVE: To evaluate the outcomes of total hip arthroplasty (THA) for coxarthrosis due to dysplasia with acetabular reconstruction of an uncemented cup in conjunction with a femoral head autograft. METHODS: A retrospective study was made on 21 hips in 20 patients (18 female and 2 male; average age, 50 years) with developmental hip dysplasia treated by THA with use of an uncemented cup. The acetabular cup was placed at the level of the true acetabulum; all patients required autogenous femoral head grafts due to acetabular deficiency. The average coverage of the acetabular cup by the femoral head autograft was 31% (range, 10% to 45%). Eight hips had less than 25% cup coverage and 13 between 25% and 50%. The average follow-up period was 4.7 years (range, 1-8 years). All patients were evaluated with the use of a modified Harris hip score. Radiographic evaluations were made by preoperative and follow-up. RESULTS: All autografts were seen to be united to host-bone. No collapse of the autograft and no hip had the evidence of loosening of component seen in all patients. According to the modified Harris hip score, the average hip score increased from 46 at preoperation to 89 at the final review. Preoperative leg-length discrepancy was greater than 2 cm seen in all except 1 patient with bilateral hip dysplasia. After surgery, only 2 of 20 patients still had a leg-length discrepancy greater than 1 cm. Three hips showed minor resorption in the lateral portion of the graft which was not supporting the cup. Three hips developed grade 1 Brooker heterotopic ossification and one had grade 2. CONCLUSIONS: THA with an uncemented cup in conjunction with a femoral head autograft for coxarthrosis due to dysplasia could obtain favorable results. This method could provide reliable acetabular fixation and appeared to restore acetabular bone stock in patients with developmental hip dysplasia when the coverage of the cementless cup by the graft does not exceed 50%. PMID- 15363241 TI - [The effect of intraoperative warming on patient core temperature]. AB - OBJECTIVE: To investigate the influence of using fluid warming and forced-air warming system on patient core temperature, blood loss, blood transfusion, extubation time, and postoperative shivering. METHODS: Forty ASA (American Society of Anesthesiologists' Physical Status) I-II patients, aged 21-69 years, scheduled for elective abdominal surgery under general anesthesia, were enrolled in the study. The patients were premedicated with intramuscular dolantin 50 mg and atropine 0.5 mg. Anesthesia was induced with midazolam 1 mg, fentanyl 50-100 microg and propofol 1.5-2.0 mg/kg. Tracheal intubation was facilitated with vecuronium 1mg and succinylcholine 1.5-2.0 mg/kg. The patients were mechanically ventilated and anesthesia was maintained with isoflurane 1.5-2.0%, 50% N2O in oxygen and intermittent iv boluses of fentanyl (total dose 5-6 microg/kg). Vecuronium was used for muscle relaxation during maintenance of anesthesia. The patients were randomly divided into 2 groups: control group (n = 20) and warming group (n = 20). In both groups, the patients were covered with surgery blanket. In the warming group, patients were additionally warmed with fluid warming device and forced-air warming system during the operation. The core temperature was recorded every 20 minutes during the operation, as well as the blood loss, blood transfusion, extubation time and postoperative shivering. RESULTS: The core temperature at the end of the surgery was (36.4 +/- 0.4) degrees C in the warming group and (35.3 +/- 0.5) degrees C in the control group. The difference was statistically significant (t = 7.547, P < 0.001). There was no significant difference of blood loss and blood transfusion between two groups. The extubation time was significantly shorter in the warming group [(18 +/- 6) vs (26 +/- 10) min, t = -3.364, P = 0.002]. 6 patients shivered postoperatively in the control group and none in the warming group (chi2 = 7.059, P = 0.008). CONCLUSION: Fluid warming system and forced-air warming system can effectively maintain normothermia during the surgery and then help to reduce the extubation time and postoperative shivering. PMID- 15363242 TI - [Effects of sevoflurane on membrane permeability of alveolar capillaries in rats with acute lung injury caused by endotoxin]. AB - OBJECTIVE: To observe the effect of sevoflurane on membrane permeability of alveolar capillaries in rats with acute lung injury and the ratio of inflammatory cells in bronchoalveolar lavage fluid in rats with acute endotoxin lung injury. METHODS: 48 Wistar rats were selected and divided into group C, L, S1L and S2L after injection evans blue 50 mg/kg in random with 12 rats in each group. Group C was taken as control group, 1.2 ml normal saline was injected into the rats via femoral vein and then the rats were mechanically ventilated for 4 hours; The rats in group L were also mechanically ventilated for 4 hours after injection of endotoxin 5 mg/kg via the same vein. For the rats in group S1L and S2L, 1.0 or 1.5 minimal alveolar concentration (MAC) sevoflurane was inhaled with mechanical ventilation after injection of endotoxin 5 mg/kg. Evans blue was not injected into 6 rats of each group in order that the 6 rats could be used for pathological examination and alveoli lavage, lung pathomorphological score of the lung, lung wet/dry weight ratio, the content of lung water, lung permeability index, content of evans blue, total amount and ratio of inflammatory cells in bronchoalveolar lavage fluid (BALF) were all determined. RESULTS: Sevoflurane of 1.0 MAC and 1.5 MAC made lung wet/dry weight ratio and content of lung water change insignificantly; lung permeability index, content of evans blue and pathomorphological score in group S1L decreased from 4.86 +/- 0.82, 112.21 +/- 11.44 ng/mg, 9.17 +/- 0.90 to 3.98 +/- 0.50, 92.85 +/- 11.80 ng/mg, 7.50 +/- 0.96; group S2L decreased to 3.91 +/- 0.34, 96.33 +/- 8.79 ng/mg, 7.67 +/- 0.75. Sevoflurane of 1.0 MAC and 1.5 MAC did not have a significantly effect on total amount and ratio of inflammatory cells in BALF. CONCLUSION: Membrane permeability of alveolar capillaries after acute endotoxin lung injury decreased by inhalation of sevoflurane of 1.0 MAC and 1.5 MAC and pathological injury of lung tissue relieved. PMID- 15363247 TI - [Rectal stromal tumors: a clinicopathological study of 16 cases]. AB - OBJECTIVE: To investigate the clinical diagnosis, treatment and immunohistochemical characteristics of rectal stromal tumors. METHODS: Immunohistochemical expression of CD117 was retrieved in 20 cases of mesenchymal tumors of the rectum. And we analyzed the immunohistochemical characteristics, clinical data of rectal stromal tumors, and the results retrospectively. RESULTS: Sixteen cases of rectal stromal tumors, 3 cases of leiomyosarcomas and 1 case of schwannoma were diagnosed. Histologically, 2 cases of GISTs were classified as benign, 1 as borderline and 13 as malignant. All of rectal stromal tumors (100%) were strongly positive for CD117, and 14 cases (88%) positive for CD34. The demographic profile of rectal stromal tumors showed a male predominance with average age of 60 years old. The main symptoms were urinary retention, constipation and abdominal pain. 14 cases were positive in digital rectal examination. The recurrence rate of local and radical resection in malignant stromal tumors was 4/4 and 3/6. 1, 3, 5-year survival rates were 89%, 64%, 48%, respectively. After operation mean survival time of was 47 months. CONCLUSIONS: The specific GIST constituted the majority of mesenchymal tumors in rectum. It usually showed malignant biological behavior. Invasion and recurrence were common. Earlier diagnosis and radical resection had better prognosis. Periodically following up can help to detect the recurrence timely. PMID- 15363248 TI - [Effecting observation of 3-dimensional conformal radiotherapy combined with chemotherapy for rectal cancer of postoperative local recurrence]. AB - OBJECTIVE: To evaluate the effects of 3-dimensional conformal radiation therapy combined with chemotherapy for rectal cancer of postoperative local recurrence. METHODS: Fifty-six rectal cancer patients of postoperative local recurrence were treated with 3-dimensional conformal radiotherapy (66 approximately 68 Gy in 2 Gy daily fraction) combined with chemotherapy (5-Fu 0.75, Days 1 - 5 and Days 29 - 33; Cisplatin 40 mg, Days 1 - 3 and Days 29 - 31). Acute and late radioreactions and survival rates were observed. Kaplan-Meier method was used for survival analysis. RESULTS: The 1, 2, 3 years local control rate was 87.5%, 66.1%, 38.2%, respectively. The 1, 2, 3 years disease-free survival and overall were 82.7%, 56.6%, 30.1% and 88.3%, 66.8%, 44.2%, respectively. The median survival was 25.3 months. Acute small enteritis were the most common but acceptable radioreactions Grade 1 approximately 2. No late radioreactions occurred. CONCLUSIONS: 3 dimensional conformal radiotherapy combined with chemotherapy can be considered as an effective and feasible approach to cancer patients of postoperative local recurrence. PMID- 15363249 TI - [Discussion for clinical treatment in early stage of colorectal carcinoma with the invasion limited in the layer of muscularis mucosae]. AB - OBJECTIVE: To emphasis the importance of recognizing and treating the early colorectal carcinoma with the invasion limited in the layer of muscularis mucosae, and discuss several questions in using the WHO's new diagnostic standards of the prestage of tumors of digestive system. METHODS: A retrospective review was made of 30 patients with invasive carcinoma limited in the layer of muscularis mucosae. All of the patients were treated between July 1986 and July 1999 in Shanghai Cancer Hospital. RESULTS: Among the 30 cases, there were 19 men and 11 women from 20 to 80 years dd (mean, 57.0). The tumor diameter was ranged from 0.8 cm to 8.0 cm (mean, 2.9 cm). 12 patients were given polypectomy while 18 patients were given radical resection. Among the patients given polypectomy, one got recurrence 3 years later, one had the metastasis of lungs 50 months later and had been dead already. Among the patients given radical resection, one had metastasis of lymph nodes peripheral to the tumor, one also had the metastasis of lungs 68 months later but is still alive now, and one had metastasis of lymph nodes in the right upper part of neck 15 months later and then was lost to follow up. CONCLUSIONS: The patients with early colorectal carcinoma with the invasion limited in the layer of muscularis mucosae could have metastasis of lymph nodes, get local recurrence after polypectomy and even have blood metastasis. Therefore great attention must be paid. Clinicians should be careful to chosen polypectomy. Meanwhile, in order to provide more information of the correct treatment, it will be better if the pathologists could give the original diagnosis at the same time when they use the WHO's new diagnostic standards of the prestage of tumors of digestive system in diagnosing the early invasive carcinoma limited in the layer of muscularis mucosae, for the concept of high-grade intraepithelial neoplasia will be used instead of it, and whether or not it is a really benign tumor, much more clinical research must be done later. PMID- 15363250 TI - [The effective analysis on clearance of pararectal lymph nodes for carcinoma of rectum]. AB - OBJECTIVE: To consider the relationship to survival rate and quality of life with pararectal lymphadenectomy for lower carcinoma of rectum. METHODS: The radical operation was performed on 780 cases of progressive cancer located at peritoneal reflection or below it, Among them, 352 cases only cleared in abdominal cavity, 428 cases coupled with extra-peritoneal histopathological type. RESULTS: Urinary function injured, the group cleared in abdominal cavity was 12 cases, accounted for 3.6%; the group coupled with extra-peritoneal clearance was 225 cases, for 52.5% (P < 0.01). Sexual function damaged (only for male), the abdominal cavity group was 23 cases, for 12.6% (23/185); the coupled group was 127 cases, for 53.4% (127/238), (P < 0.01). Local relapse rate, the abdominal cavity group was 15.8% (56/352); the coupled group was 8.6% (41/428), (P < 0.05). 5-year survival rate, the abdominal cavity group was 52.2%; the coupled group was 58.5% (P < 0.05). CONCLUSION: By contrast, although abdominal cavity coupled with extraperitoneal lymphadenectomy acted to cut down local relapse and to elevate 5 year survival rate, the postoperative quality of life appeared to be seriously affected. PMID- 15363251 TI - [Radical resection with total mesorectal excision under the laparoscope in the treatment of rectal cancer]. AB - OBJECTIVE: To evaluate the effect of radical resection with total mesorectal excision under the laparoscope in the treatment of rectal cancer. METHODS: Under the laparoscope and with the ultrasonic scalpel, total mesorectal excision in 38 patients with rectal cancer was performed. In operation, alone the parietal layer of pelvic fascia and inside the autonomic nerve trunk, the mesorectum was excised. All patients were followed up for 2 years. RESULTS: All cases were underwent successfully and no complication was observed. Postoperative flatus and feces were passed at 32 hours and postoperational hospital stay were 7.5 days. No trocar port implantation and no death case was observed. Two local recurrence developed in 2 years follow-up. CONCLUSIONS: Radical resection with TME is a new progress of less damageable operation, which has advantages of low postoperative local recurrence rate and help for pelvic autonomic nerve. PMID- 15363252 TI - [Germline E-cadherin gene mutation screening in familial gastric cancer kindreds]. AB - OBJECTIVE: To evaluate the frequency and nature of E-cadherin gene (CDH1) germline mutations in familial gastric cancer kindreds of china. METHODS: Five familial gastric cancer kindreds of Chinese origin were screened for germline CDH1 mutations, all of them meeting the clinical criteria for hereditary diffuse gastric cancer (HDGC), by PCR-DHPLC and direct sequencing. RESULTS: A new truncating mutation in exon 10 in B family was identified, producing a stop codon at position 503 (Q503X), resulting in a truncated protein. The proband of this family had metachronous development of lobular breast and diffuse type gastric carcinoma. No protein expression was detected in the lobular breast carcinoma, indicating complete inactivation of the gene. CONCLUSIONS: A Chinese gastric cancer family with CDH1 germline truncating mutation is described for the first time, and our findings suggest that lobular breast carcinoma might be part of the tumor spectrum of HDGC. PMID- 15363253 TI - [Comparing the effect of adjuvant chemotherapy by portal vein infusion with intraluminal chemotherapy for colorectal cancer]. AB - OBJECTIVE: To compare the effect of 5-fluorouracil (5-FU) portal vein infusion (PVI) for 7 days after radical resection, with intraluminal chemotherapy during operation for eliminating liver metastasis and elevating long-term prognosis in colorectal cancer. METHODS: 162 colorectal cancer patients with radical resection were divided into portal vein chemotherapy group (group A, 82 cases) and intraluminal chemotherapy group (group B, 80 cases) randomly. In group A, 5 fluorouracil were infused with 1g per day constantly for 7 days after operation through portal vein catheters, which placed into greater omental vein and fixed on the abdominal wall. In group B, intraluminal chemotherapy was given and 5 fluorouracil 0.5 g was injected into the greater omental vein during operation. RESULTS: The short-term complications and long-term effect in the two groups were compared by statistical software SPSS 8.0. Group A had more operative complications, and no statistical differences was found in hospital time and survival rate of the two groups. The 5-year survival rate is 76.7% (group A: 74.3%, group B: 79.2%), and the liver metastasis rate is 19.8%. There is no significant difference between the two group-survival curves. Multiple variable analysis suggested that Dukes' stage was the prognosis factor (P < 0.05). CONCLUSIONS: The present study demonstrated that the two chemotherapy methods play an important role in preventing liver metastasis and improving the survival rate, and the intraluminal chemotherapy would be easier and simpler. The result should be further improved by using combined chemotherapy. PMID- 15363254 TI - [The effect of pre-operative chemotherapy on calpain in cells of gastrointestinal malignant tumors]. AB - OBJECTIVE: Primary to search the rule of pre-operative chemotherapy and suitable duration for it by investigating the changes of calpain content and activity after 5'-deoxy-5-fluorouridine (5'-DFUR) per oral administered pre-operatively in different time. Further to investigate the mechanism of chemotherapy. METHODS: Seventy-three patients with gastrointestinal malignant tumors were divided into 4 groups by the time of 5'-DFUR (600 approximately 1200 mg/d) by oral administration before operation, group A, 3 days, 27 cases; group B, 1 week, 22 cases; group C, 2 weeks, 15 cases; group D, 2 months, 9 cases. And group E, control group, had 24 patients with gastrointestinal malignant tumors at the same term. The patients above all had not received the other chemotherapy and radiotherapy. Western blot and immunoelectron microscopy were employed to detect the expressing levels and activities of calpain in tumor tissues of different groups. RESULTS: Electronic microscopic examination showed gold granula mainly on the membranes of mitochondria of tumor cells to groups after chemotherapy. And the tumor cells of group A were mildly damaged. Besides that, serious injury for tumor cells of group B could be seen, and the phenomena were common in group C. But the damages to tumor cells of group D were mainly about mildness. The results of immunoelectron microscopy revealed that the contents of calpain increased following the time of chemotherapy prolonging, and peaked in group C. Still more, there was no significant difference for the results between group C and group D. The changes of calpain activities observed by western blot had the same tendency as the results from immunoelectron microscopy (r = 0.86, P < 0.0001). CONCLUSIONS: 5'-DFUR via oral administered pre-operation could have anti-cancer effect through calpain. And the effect might be strongest in 2 weeks also after chemotherapy. PMID- 15363255 TI - [Analysis of rejection after simultaneous pancreas-kidney transplantation]. AB - OBJECTIVE: To explore methods of preventing and reversing rejection after simultaneous pancreas-kidney transplantation (SPK). METHODS: Seventeen patients performed SPK operation from Sep, 1999 to Sep, 2003 were reviewed retrospectively. Immunosuppression was achieved by triple regimen consisting of cyclosporine, mycophenolate mofetil (MMF)/azathioprine and steroid. 2 patients were treated with Dalizumab, the other three patients used OKT3 as immune induction. RESULTS: 1 patient experienced the accelerated rejection, the pancreas and kidney grafts were resected because of failure of conservative therapy. 8 patients experienced renal acute rejection, 2 cases suffered from pancreas acute rejection at the same time. All these patients received daily high dose pulse steroid for 3 days. OKT3 was administered in 2 patients with steroid resistance rejection. All the grafts were successfully rescued. CONCLUSIONS: Reasonable application of immunosuppression after SPK operation and adoption of systemic measures which can reduce sensitivity of high risk receptor before SPK operation are the effective methods of preventing and treating rejection. PMID- 15363256 TI - [The changes of Oddi sphincter motility after canine pancreas transplantation with bladder drainage]. AB - OBJECTIVE: To study the changes of canine Oddi sphincter (SO) function after pancreas transplantation with bladder drainage and the effect on the graft function. METHODS: Normal canine SO, transplant canine SO and canine SO in vitro manometry were performed by triple lumen catheter. At the same time, pancreas endocrine and exocrine function after transplantation were determined. After transplantation, anti-reflux function of graft SO was also measured. RESULTS: Endocrine and exocrine function of all the transplanted dogs showed that pancreas graft function was good. Basal pressure of SO in control group was (18.5 +/- 2.8) mm Hg (1 mm Hg = 0.133 kPa). The contraction frequency was (9.7 +/- 1.5) per min, the contraction amplitude was (47.1 +/- 5.5) mm Hg, the motility index was (236 +/- 56). After transplantation, basal pressure increased to (27.8 +/- 2.8) mm Hg, frequency increased to (13.1 +/- 1.9) per min, amplitude decreased significantly to (8.3 +/- 1.8) mm Hg. There was no significant difference of motility index. Basal pressure of SO in vitro increased significantly to (37.2 +/- 5.1) mm Hg. Phasic contraction was not absent. After transplantation, the pressure in the bile duct residual did not increase in accordance with the increase of bladder pressure. CONCLUSIONS: After pancreas transplantation with bladder drainage, Basal pressure and frequency of canine SO could increase while amplitude could decrease, which provide the anti-reflux function of graft SO and may serve as an obstacle to pancreatic juice flow. PMID- 15363257 TI - [Improvement of massive human islet isolation techniques and the evaluation of isolated human islets]. AB - OBJECTIVE: To obtain massive human pancreatic islets with modified techniques and evaluation of the islets for the clinical allo-transplantation to treat type I and II diabetes. METHODS: 28 consecutive adult human pancreata were isolated with modified automated techniques. Islets were purified using continuous density gradient. The islet yield was counted with international standard known as islet equivalent (IEQ). The function of the isolated islets was evaluated by measuring DNA/insulin ratio, static glucose stimulating test in vitro and transplanting the islets into diabetic nude mice in vivo followed by abdominal glucose tolerance test and C peptide measurement. RESULTS: The yield of 28 consecutive human pancreata isolations ranged from 5 000 to 1 030 000 IEQs/pancreas with the average of 291 635 IEQs/pancreas. The first 13 isolations yielded 49 123 IEQs/pancreas, 846 IEQs/g and, purity 87% in average. The remained 15 isolations after the modifications yielded 501 813 IEQs/pancreas, 7 003 IEQs/g and purity 89% in average. The results of in vitro SGS showed good response to the different glucose concentration. 34 diabetic nude mice were transplanted under the renal capsule with the freshly isolated islets. 29 out of 34 diabetic mice obtained normoglycemia within 12 hours and the glucose tolerance tests were near normal. Serum C peptide level of transplanted mice is close to that of the control group. CONCLUSIONS: Massive human islets can be isolated with the modified techniques. Quality assessment of these islets both in vitro and in vivo has indicated that these high quality human islets could be used for the clinical allogeneic islet transplantation. PMID- 15363258 TI - [The protective role of gadolinium chloride in lung injury associated with acute necrotizing pancreatitis]. AB - OBJECTIVE: To discuss the role of gadolinium chloride (GdCl(3)) in lung injury associated with acute necrotizing pancreatitis (ANP). METHODS: Experimental animals were randomized into five groups (n = 18 for each group): normal control group, ANP group, GdCl(3) pretreatment group, ANP GdCl(3) pretreatment group, ANP GdCl(3) treatment group. Rat ANP model was induced by intraductal administration of 3% sodium taurocholate. Alveolar macrophages (AM) were obtained by bronchoalveolar lavage. The blood gas assay, the ratio of wet/dry tissue, protein content of bronchoalveolar lavage fluids (BALF), the myeloperoxidase (MPO) of lung tissue and generation of TNFalpha and NO by AM were evaluated. The apoptosis of AM was checked by agarose gel electrophoresis analysis, transmission electric microscopy observation and cytometry propidium iodide single stained method. The lung tissue was examined by histology. RESULTS: The parameter of GdCl(3) pretreatment group compared with normal control group had no statistical significance (P > 0.05). The indicators of ANP GdCl(3) pretreatment group and ANP GdCl(3) treatment group were elevated compared with the normal control group and had statistical significance (P < 0.05). But compared to the ANP group, they were all decreased and also had the statistical significance (P < 0.05). The 180 - 200 bp ladder pattern unique to apoptosis in agarose gel electrophoresis and the apoptotic typical morphologic feature in AM by transmission electric microscopy and typical subdiploid peak in DNA content figure could be observed in ANP GdCl(3) pretreatment group and ANP GdCl(3) treatment group, while the other three groups could not. CONCLUSIONS: Lung injury associated with ANP could be ameliorated by application of GdCl(3) through inducing apoptosis of AM of ANP. PMID- 15363259 TI - [Portal venous and enteric drainage in simultaneous pancreas kidney transplantation]. AB - OBJECTIVE: To summarize the initial experience of simultaneous pancreas kidney transplantation (SPK) with portal venous and enteric drainage. METHODS: Between Jane 2001 and Jane 2003, SPK were performed in 5 patients. Systemic venous enteric drainage (SED) was used in the first 2 patients and portal venous-enteric drainage (PED) in the last 3 cases. All patient were immunosuppressed with quadruple therapy, which included anti-CD25 mAb (Zenapax/Simulect) induction therapy, FK506, mycophenolate mofetil (MMF), and prednisone baseline therapy. The complications were analyzed. RESULTS: Serum glucose and renal function of the 5 cases were normal and no further insulin was needed within 7 days post-operation. No technique complications such as duodenal fistula and thrombosis were observed, One episode of acute rejection of kidney allograft occurred in one patient with SED, and resolved with a bolus corticosteroids. One case with SED and one with PED were died of sepsis and FK506 toxicity 4 weeks after transplantation. The death occurred with functioning pancreas graft. No latter complications were observed in the 3 survived patients with excellent graft functions. CONCLUSIONS: Both methods of SED and PED can be performed successfully and with no latter complications. But with its potential physiologic and immunologic advantages, PED might be a standard procedure for SPK. PMID- 15363260 TI - [Clinical research on radiofrequency ablation for hypersplenism]. AB - OBJECTIVE: To study the security and curative effect of radiofrequency ablation (RFA) for hypersplenism due to portal hypertension in the near future. METHODS: 12 patients of the disease were treated with RFA. The blood count showed that WBC is (2.0 +/- 0.7) x 10(9)/L, PLT (25 +/- 10) x 10(9)/L, RBC (3.08 +/- 0.56) x 10(9)/L. In manners of systemic anaesthesia, through skin or in the operation or under the celioscopes the RFA needles are inserted the upper and middle parts of spleen in the instructing of US. Average time of RFA is (36.4 +/- 5.4) s. RESULTS: After the RFA, 1 case develop hematoma under the envelope of liver, and 1 case developed skin burn; 6 patients appeared middle or little fluid in the left thorax, 2 of them were drawn through puncture. The volume of spleen and damaged part of spleen from RFA were determined by enhanced CT in the days of 10 after RFA, the percentage of damaged part of spleen from RFA is 49.1% (23.3% - 88.9%). In the days of 14 after RFA, RBC is (5.5 +/- 0.9) x 10(9)/L, PLT is (124 +/- 21.36) x 10(9)/L; In the days of 33 after RFA, RBC is (5.4 +/- 1.1) x 10(9)/L, PLT is (205 +/- 34) x 10(9)/L. The levels of WBC and PLT in the blood after RFA are significantly higher than those of levels of WBC and PLT before RFA (P < 0.01). CONCLUSIONS: Radiofrequency ablation for Hypersplenism due to portal hypertension of hepatitis hepatocirrhosis is safety and credibility and has excellent curative effect in the near future. PMID- 15363270 TI - [Effects of escharectomy during shock stage on tissue high mobility group box-1 expression and balance of pro-/anti-inflammatory response in rats after severe thermal injury]. AB - OBJECTIVE: To investigate the effects of escharectomy during shock stage on tissue high mobility group box-1 protein (HMGB1) expression and balance of pro /anti-inflammatory cytokines, and to elucidate the potential mechanism underlying beneficial effect of early escharectomy after severe burns. METHODS: Wistar rats inflicted by 30% full-thickness thermal injury were randomly divided into thermal injury group, 24 h escharectomy group and 72 h escharectomy group, in which escharectomy were performed at 24 and 72 h postburn, respectively. Gene expression of HMGB1, interleukin-10 (IL-10), and tumor necrosis factor-alpha (TNF alpha) in liver and lungs was detected with reverse-transcription PCR, and protein levels of IL-10 and TNF-alpha in liver and lung tissues were measured by ELISA. The plasma AST and ALT contents, and pulmonary myeloperoxidase (MPO) activity were also assayed. RESULTS: The mRNA expression of HMGB1 and TNF-alpha in liver and lungs was up-regulated on postburn day 2, with IL-10 over-expression on postburn day 8. In the 24 h escharectomy group, HMGB1 and TNF-alpha mRNA expression in liver and lungs was down-regulated on postburn day 4, and IL-10 expression returned to normal range on postburn day 8, while the down-regulation of HMGB1, TNF-alpha and IL-10 were not noted in the 72 h escharectomy group. There were two peaks in liver TNF-alpha protein levels appearing on postburn days 2 and 8, respectively, with an unexpected marked decrease on day 4 in thermal injury controls, yet liver TNF-alpha levels maintained in normal range in animals of 24 h and 72 h escharectomy groups. The ratios of TNF-alpha to IL-10 protein levels in liver tissue were significantly increased on postburn days 2 and 4 (P = 0.0001 and 0.002, respectively), while escharectomy during shock stage markedly reduced hepatic TNF-alpha to IL-10 ratios (P = 0.0008 and 0.040, respectively). No significant changes in TNF-alpha protein levels in lung tissue were observed. Additionally, plasma AST as well as ALT contents, and pulmonary MPO activity were markedly decreased on postburn days 4 and 8 in the 24 h escharectomy group compared to the 72 h escharectomy group or thermal injury controls (P < 0.05). CONCLUSIONS: Escharectomy during burn shock stage could inhibit the over expression of both early and late inflammatory mediators, and maintain the balance of pro-/anti-inflammatory response, thereby improving multiple organ functions in rats following severe burns. PMID- 15363271 TI - [A study of the abnormalities of human epiderm in keloids and hypertrophic scars]. AB - OBJECTIVE: To investigate the abnormalities of human epiderm in keloids and hypertrophic scars. METHODS: Biopsies from ten untreated keloids (duration of disease 3 - 30 years) and ten hypertrophic scars (duration of disease 6 - 10 months) and five normal adult skin specimens. Total RNA was isolated from normal adult skin. A cDNA fragment (base 5941 - 6481 bp) of the full-length human Tenascin-C cDNA was synthesized by polymerase chain reaction and subcloned in pGEM-T-easy. Dioxigen-labeled anti-sense and sense probes were synthesized by using a Sp6/T7 in vitro RNA synthesis kit in the present of Dig-UTP. In situ hybridization was performed on 4% paraformaldehyde-fixed and wax-embedded sections of keloids and hypertrophic scars. NBT-NCIP was used in color detection. Immunohistochemical procedure. The sections were incubated with antibodies (anti Tenascin-C, anti-CK-16 and anti-Ki-67). Ultrasensitive Streptavidin Peroxidase staining was performed following established procedures. RESULTS: The study show that the proliferation of epidermal keratinocytes in keloids and hypertrophic scars is very clear. The expressions of Tenascin-C mRNA in keloids epidermal keratinocytes markedly increased in contrast with epidermal keratinocytes of hypertrophic scars and adult skin. The CK-16 and Ki-67 stainings significantly enhanced in the epidermal keratinocytes of keloids and hypertrophic scars. CONCLUSIONS: The different expressions of Tenascin-C, CK-16 and Ki-67 among normal adult skin, keloids and hypertrophic scars show the abnormalities of epidermal keratinocytes proliferation and differentiation in keloids and hypertrophic scars. PMID- 15363272 TI - [Treatment of paraneoplastic pemphigus with Castleman's disease]. AB - OBJECTIVE: To discuss the clinical findings and treatment of paraneoplastic pemphigus (PNP) with Castleman's disease. METHODS: To investigate the clinical, histopathologic and CT findings of 8 cases paraneoplastic pemphigus with Castleman's disease. RESULTS: All of 8 patients were diagnosed PNP first and were found Castleman's tumor incidently during routine examination. All 8 cases showed severe erosion or ulcer of the oral mucosa with various skin lesions. Histopathologically, there were intraepidermal acantholytic vesicle, basal cell liquefaction, necrotic keratinocytes in the epidermis and lymphocyte infiltration in the upper dermis. CT scan appeared solitary mass in these patients. Some of them were attacked by bronchiolitis obliterans. All 8 patients were failed by use of predisone. Obvious relief of PNP and pulmonary lesion occurred after tumor was rescted. CONCLUSIONS: Paraneoplastic pemphigus with Castleman's disease is a rare disease. The key step is to find and resect the tumor in abdomen. CT scan should be used to detect the tumor in patients with PNP, especially, when predisone was failed in treatment. PMID- 15363273 TI - [Study on distribution of interstitial cells of Cajal in the sigmoid colon of patients with slow transit constipation]. AB - OBJECTIVE: Slow transit constipation (STC) is a colonic motor disorder whose etiology remains unclear. Recent studies have demonstrated a crucial role for interstitial cells of Cajal (ICC) in regulation of intestinal motility. The aim of this study was to examine the distribution of ICC within the normal sigmoid colon and STC patients. METHODS: Twelve patients with STC and eight age-matched controls were studied. ICC were identified with a monoclonal antibody to c-kit by an indirect immunofluorescence method. Immunostained tissues were examined with a laser scanning confocal microscope and the area occupied by ICC was calculated with image analysis software. RESULTS: ICC were located in the external muscle layers including longitudinal muscle (LM), myenteric plexus (MP), circular muscle (CM) and submucosal border (SMB). Two types of Kit-positive ICC were observed: bipolar cells characterized by one or two long processes, and multipolar cells with long stellate processes extending in various directions. A higher percentage of ICC was present in the MP regions and CM layers compared with the SMB and LM layers. Tissues from STC patients showed considerably decreased in number of ICC located in the four regions (ICC-LM, ICC-MP, ICC-CM, ICC-SMP), especially for ICC SMP, almost completely disappeared. CONCLUSION: Decreased c-kit + ICC in number may play an important role in the pathophysiology of STC. It remains to be determined whether loss of ICC is primary or secondary to another lesion. PMID- 15363274 TI - [The anatomic study of chyle leakage due to operation on abdominal region]. AB - OBJECTIVE: To provide morphological basis for chyle leakage due to operation on upper abdomen or retroperitoneum region. METHODS: The original part of thoracic duct, cisterna chyle, intestinal trunk, left and right lumbar trunks were examined in 32 adult cadavers. RESULTS: (1) The occurrence rate of cisterna chili was 22% (7 cases), among which 4 cases were oval, 3 cases were triangle. The cisterna chyle was (24 +/- 6) mm in length; the width of middle part was (4.1 +/- 0.9) mm. It was located to the right of midline at the level between the twelfth thoracic vertebral body and the second lumbar vertebral body anteriorly. (2) The original part of thoracic duct was (2.8 +/- 0.7) mm in diameter. The confluence form of thoracic duct included: left lumbar trunk and intestinal trunk united to form the common trunk first, right lumbar trunk then joined the common trunk (9 cases, 36%); right lumbar trunk and intestinal trunk united to form the common trunk first, left lumbar trunk then joined the common trunk (8 cases, 32%); left and right lumbar trunk united to form the common trunk first, intestinal trunk then joined the common trunk (4 cases, 16%); left, right lumbar trunk and intestinal trunk joined together (3 cases, 12%). (3) The intestinal trunk was (36 +/- 15) mm in length. It ascended on the left of descending aorta, superior to the left renal artery, crossed the second lumbar vertebra anteriorly, and joined left or right lumbar trunk to form common trunk, which extended to the cisterna chili or thoracic duct to the right of lumbar vertebra. (4) The lengths of left and right lumbar trunks were (107 +/- 24) mm and (111 +/- 18) mm, the external diameters of origins were (1.7 +/- 0.4) mm and (1.9 +/- 0.4) mm, and the external diameters of terminations were (2.2 +/- 0.6) mm and (2.2 +/- 0.5) mm, respectively. CONCLUSION: The larger lymph tubes should be protected emphatically in the relevant region when dissecting the root of celiac and superior mesenteric artery and the termination of inferior mesenteric vein during abdominal operation. PMID- 15363276 TI - [The prognosis of the total proctocolectomy and ileal-pouch anal anastomosis]. AB - OBJECTIVE: To evaluate the prognosis of the total proctocolectomy and ileal-pouch anal anastomosis (IPAA) for ulcerative colitis (UC) and familial adenomatous polyposis (FAP). METHODS: Sixty-one patients with ulcer colitis or familial adenomatous polyposis were performed total proctocolectomy and ileal pouches-anal anastomosis during 1985 to 2002. There are S type pouch 25 cases, S-J type pouch 13 cases, J type pouch 17 cases and W type pouch 6 cases. The complication and function after the IPAA were also discussed. RESULTS: No patient died after operation. The total morbidity is 16% (10/61), the morbidity of group UC (6/25) is higher than FAP's (4/34). The W type pouch's morbidity is higher than other three types', the operation with stapled technique is associated with fewer complication than hand-sewn IPAA (2/20 vs 8/41), however, there is also no significant difference between them. The number of stools per 24 hours is 4.2, the percent of the normal continence of daytime and nighttime is 84% (43/51) and 75% (38/51) respectively. There's only about 6% (3/51) patient with fecal incontinence. The most patients are satisfied with IPAA. CONCLUSION: The proctocolectomy ileal pouch-anal anastomosis for FAP and UC has few complication with accepted frequency and preserve a good anal function, it is an ideal alternative approach. PMID- 15363277 TI - [Clinical significance of expression of VEGF and bFGF in thyroid carcinoma]. AB - OBJECTIVE: To study the clinical significance of expression of VEGF and bFGF in thyroid carcinoma. METHODS: Immunohistochemical SP technic was used to investigate the expression of VEGF and bFGF in 90 cases of papillary carcinoma, 14 cases of undifferentiated carcinoma, 10 cases of normal thyroid tissue. RESULTS: The positive rate of VEGF and bFGF in thyroid carcinoma was 63.5% and 59.6% respectively, Which was significantly higher than that in normal thyroid tissue (P < 0.01); The positive rate of VEGF and bFGF in undifferentiated carcinoma was 92.9% and 85.7% respectively, which was significantly higher than that in papillary carcinoma (P < 0.05); The positive rate of expression of VEGF and bFGF was correlated with lymph gland metastasis (P < 0.05); A positive correlation between these two expresses existed in thyroid carcinoma (gamma = 0.596, P < 0.05). CONCLUSION: VEGF and bFGF can be regarded as a parameter to evaluate the biological behavior and prognosis of thyroid carcinoma. PMID- 15363278 TI - [Whether VI region lymph nodes belong to primary site of the thyroid carcinoma or lateral cervical lymph node metastases]. AB - OBJECTIVE: To discuss the evaluation of elective neck dissection (END) for the cN(0) patients with papillary thyroid carcinoma (PTC). METHODS: By analyzing the recurrent and metastatic region (thyroid, group VI lymph nodes, lateral neck region, beyond neck) of 139 PTC patients treated secondly in our hospital, group VI lymph nodal metastasis is divided into recurrence of primary site and distinguished from lateral neck lymph nodes. The clinical value of END for cN(0) PTC patients is also retrospectively analyzed. RESULTS: Thyroidal recurrence accounts for 83% (73/88). Level VI metastasis accounts for 76% (67/88), 17 patients have received END and account for 65% (17/26). Metastasis to lateral neck lymph nodes account for 17% (17/98), among these patients, 5 patients have received END and account for 19% (5/26). CONCLUSIONS: For the patients with cN(0) PTC, ipsilateral thyroid lobectomy plus level VI dissection is recommended and lateral END (level II-V) is not supported. For the patients with cN(0) but UB N(+) and/or CT N(+), lateral END (level II-V) is recommended. The necessity of CT examination in the diagnosis and treatment of thyroid cancer should be emphasized. PMID- 15363279 TI - [The effect of endothelin receptor in androgen-independent prostate cancer]. AB - OBJECTIVE: To study the expression of ET receptor and the apoptosis after intervened with ET receptor antagonist in androgen-independent prostate cancer. METHODS: PC3, an androgen-independent prostate cancer cell line, was used. The expression of ETA and ETB receptor in PC3 was measured through RT-PCR. After intervened with selective ETA and ETB receptor antagonist, the apoptosis in PC3 was studied through flow cytometry and electron microscope. RESULTS: Clear signal was obtained in PC3 for ETA receptor mRNA transcript, while the signal for ETB receptor mRNA transcript was very weak. The expression of ETA receptor mRNA was obviously reduced and the apoptosis of PC3 cell was observed after intervened with selective ETA receptor antagonist. There was no change after intervened with selective ETB receptor antagonist. CONCLUSION: ET-1 exerts its effects through the ETA receptor subtype and ETB receptor is silenced in PC3. The expression of ETA was reduced and the apoptosis was observed in PC3 when ETA receptor was blocked. It was dose-dependent. PMID- 15363280 TI - [Change of nerve growth factor mRNA in human detrusor in bladder outlet obstruction with benign prostatic hyperplasia and their implication]. AB - OBJECTIVE: To investigate the change of nerve growth factor (NGF) mRNA in human detrusor in bladder outlet obstruction (BOO) with benign prostatic hyperplasia (BPH) and their implication. METHODS: Eight cases of bladder cancer and 40 patients with BPH were included in this study. All patients were divided into three groups, a control group, an obstructive detrusor stability group and an obstructive detrusor instability group. NGF mRNA in detrusor from all patients was measured using a RT-PCR. RESULTS: The RT-PCR study indicated that NGF mRNA was expressed in detrusor of three groups of patients. There were significant differences among the three groups (P < 0.01). The expression of NGF mRNA in the obstructive instability group was higher than that in the obstructive stability group and in the control group. The NGF mRNA level in the obstructive stability group was higher than that in the control group. CONCLUSION: The expression of NGF mRNA in detrusor was elevated in BOO with BPH. The elevated expression of NGF mRNA might be correlated with detrusor instability (DI) due to BOO. PMID- 15363281 TI - [Effects of ulinastatin on renal ultrastructure after ischemia-reperfusion in rats]. AB - OBJECTIVE: To investigate the effect of ulinastatin on renal function and ultrastructure changes after renal ischemia-reperfusion in rats. METHODS: Acute ischemic renal injury model was established (45 min of bilateral renal ischemia and reperfusion for 24 h). Thirty Male SD rats were randomly divided into 3 groups: sham operation group (control group or group C, without renal ischemia), renal ischemia-reperfusion group (ischemia-reperfusion group or group I, without ulinastatin), renal ischemia-reperfusion and ulinastatin intravenous injection group (ulinastatin group or group U). BUN level, serum creatinine values and renal ultrastructure were measured. RESULTS: Serum creatinine (167 +/- 39) micromol/L and BUN concentration (21 +/- 7) mmol/L in group I were significantly higher than those in group U: serum creatinine (116 +/- 13) micromol/L and BUN concentration (14.1 +/- 2.6) mmol/L (P < 0.05). The renal ultrastructure was greatly injured in group I, meanwhile, it was obviously ameliorated in group U. CONCLUSION: Ulinastatin greatly improved renal function and provides remarkable protection on renal ultrastructure after ischemia-reperfusion of kidney in rats. PMID- 15363291 TI - [Microsurgical resection of the recurrent craniopharyngiomas]. AB - OBJECTIVE: To study the effective method of microsurgical resection of the recurrent craniopharyngiomas. METHODS: Microsurgical resection underwent in 40 cases with the recurrent craniopharyngiomas that accounted for 24 men and 16 women with mean age 35.1 year old. Visual deterioration was mainly complained in 25 patents, headache of symptoms in 9 patients, defect of visual field in 2 patients, amenorrhea in 2 patients, hyposexuality in one patient and diabetes insipidus in the one. Average history was 2.9 years. The superior sellar tumors on MR imaging grew in 19 cases, the superior-inferior sellar mass in 9 cases, intra-sellar in 5 cases and the tumors into the third ventricle in 7 cases. The huge calcification tumors were found in 5 cases, cystic tumors in 21 cases, and solid tumors 2 cases. Hydrocephalus presented in 12 cases. Evolution of tumor residuum was revealed in 31 cases after initial surgery and recurrent tumor after completing total removal of the tumor in 9 cases. The pterional approach was employed in 33 cases. The longitudinal fissure to the corpus callus approach in 2 cases was available for resection of the third ventricular tumor through the fornix column and septum pellucidum spaces. RESULTS: In 33 cases with the pterional approaches, total removal of the tumors were completed in 22 cases, subtotal removal of tumors in 9 cases, and partial removal in 2 cases. In 5 cases with subfrontal approach, 4 patients the total removal of tumors obtained in 4 cases and one subtotal removal of tumor in one. Of 2 cases with the longitudinal fissure to the corpus callus approaches, one case was achieved with the total removal of tumor and the other with subtotal removal of tumor. The pituitary stalk was preserved in 8 cases, the pituitary stalk was severed in 9 cases and the pituitary stalk was not identified in 23 cases when surgery. 17 patients experienced diabetes insipidus and 12 patients had the hypothalamic hypofunction after surgery. One death occurred from the hypothalamic hypofunction 35 day after surgery. By follow-up from 3 months to 3 years, 22 patients returned normal life, 11 patients can carry out their self-life, and 6 patients need care. CONCLUSIONS: The desirable removal of recurrent craniopharyngioma could be completed in the majority of patients although the reoperation of the tumors was performed very difficulty owing to the tumor adhesive to the surrounding hypothalamic structures. PMID- 15363292 TI - [Jugular foramen schwannomas: a review of 17 cases]. AB - OBJECTIVE: To analysis the clinical presentation, radiological findings, surgical techniques and outcomes of jugular foramen (JF) schwannomas. METHODS: We reviewed our 10-year experience in the surgical treatment of 17 patients suffered from JF schwannomas in Hua Shan Hospital, Shanghai. A total of 8 males and 9 females with a mean age of 42 years underwent surgical procedures. A relative long period of 53-month symptomatic history was shown before surgery. The main clinical presentation are vertigo and hearing difficulty in 10 cases, atrophy of unilateral muscles of tongue in 9 cases, involvement of lower cranial nerve in 8 cases. The classification of tumors was type A (at cerebellopontine angle with minimal enlargement of the JF) in five cases, type B (JF with intracranial extension) in 3 cases, type C (extracranial tumors with JF extension) in 2 cases and type D (dumbbell-shaped with both intra-and extracranial components) in 7 cases. RESULTS: Far lateral approaches were used in 10 cases, retrosigmoid suboccipital approaches were used in 5 cases. Submandibular approaches were selected in other 2 cases. Gross total removal was achieved in 12 cases, and subtotal removal in 5 cases. Follow-up revealed marked improvement from preoperative symptoms in 9 cases and no additional deficits in 3 cases. 5 cases suffered from additional neurological deficits. There were two cases of temporary hoarseness and gradually improved within follow-up. Two patients suffered from swallowing problems as a new deficit. One patient had facial palsy. CONCLUSIONS: JF schwannomas can be surgically treated with relative good outcomes. Surgical approaches should be tailored according to the tumor extension. PMID- 15363293 TI - [The diagnosis and microsurgical treatment of solid cerebellar hemangioreticuloma]. AB - OBJECTIVE: To study the clinical features, neuroimaging presentations, diagnosis and differential diagnosis, and operative warning events of the solid cerebellar hemangioblastoma. METHODS: The clinical data of 48 patients with solid cerebellar haemangioreticuloma were retrospectively analysed. Preoperative examinations included CT and MRI in all patients, DSA in 20, preoperative embolization of feeding arteries of tumor in 16, a preoperative ventriculoperitoneal shunt was placed in 5 cases. The lesion distribution of 48 patients was as follows: 14 lesions were situated in the left hemisphere of cerebellum with an extent into the cerebellopontine angle in 2 cases, 12 in right hemisphere of cerebellum, 8 in superior vermis, 7 in inferior vermis, 6 in left subtentorium, 1 in left cerebellar tonsil. RESULTS: The tumors of 48 patients were totally removed. One patient occurred normal perfusion pressure breakthrough during surgery, and gastrointestinal hemorrhage on the second day after operation. Other patients all were intra- and postoperatively uneventful. Except for autoinfusion, no patients transfused heterogenous blood. There were no operative mortality and serious complications in this series. CONCLUSIONS: The solid cerebellar hemangioblastoma was benign neoplasm. Abiding by properly operative techniques, the optimal results can be obtained. PMID- 15363294 TI - [Surgical treatment of gliomas involving the supplementary motor area in the superior frontal gyrus]. AB - OBJECTIVE: To explore surgical treatment of gliomas involving the supplementary motor area (SMA) in the superior frontal gyrus. METHODS: Clinical data and follow up outcome of 16 patients with low graded astrocytomas involving the supplementary motor area were analyzed. RESULTS: SMA syndrome was developed in 6 patients in whom the posterior tumor resection line was at a distance of more than 1 cm from the precentral sulcus and resolved after 12 months. Hemiplegia occurred however in 8 patients in whom the resection line was less than 1 cm to precentral sulcus and only resolved in 3 patients during follow period 12 months. CONCLUSIONS: When the resection is performed at a distance of less than 1 cm from the precentral sulcus, surgery for gliomas of involving the supplementary motor area in the superior frontal gyrus may be result in permanent morbidity. PMID- 15363295 TI - [Diagnosis of intracranial malignant melanoma]. AB - OBJECTIVE: To analyse the diagnosis points of intracranial malignant melanoma. METHODS: There were ten patients with intracranial malignant melanoma (but without clear extracranial primary lesion), whose epidemiology, clinical symptom, physical sign and imaging data were analysed. RESULTS: Most of patients were young men, The Clinical symptom or physical sign are similar to brain tumor or cerebral hemorrhage, but the course was shot, two cases of among having skin melanotic nevus. As for most of cases, CT were high density, MRI T1WI showed high signal and T2WI low signal, which often had hemorrhage. Histological examination showed all cases were malignant melanoma. Preoperative misdiagnosis was up to eight cases (80%). CONCLUSIONS: For preoperative accurate diagnosis, this disease's epidemiology, clinical symptom, extracranial physical sign and imaging data points could be synthetical analysed. PMID- 15363296 TI - [Experimental research of myogenic motor evoked potentials to transcranial magnetic stimulation for spinal cord monitoring]. AB - OBJECTIVE: To evaluate the validity of intraoperative magnetic MEP (motor evoked potentials) monitoring in a spinal-cord-menaced surgery. METHODS: 32 rabbits were employed in weight-drop spinal cord contusion model. After anesthetized with a combination of Ketamine and Droperidol the spinal cords were surgically exposed with the dura intact, and the contusion injuries were delivered except the rabbits in control group. The MEPs were recorded and the relationship between the variation of the MEPs and the residual locomotor capacity after spinal cord injury was analyzed. RESULTS: The 6 rabbits in mild-spinal-cord-injury group experienced transient attenuation of their TMS-MEPs, and the locomotor capacity remained intact (scores of 5) in almost all rabbits (5 of 6) when assessed 24 hours later; In the moderate-spinal-cord-injury group the 8 rabbits lost their TMS-MEP immediately after the weight-drop contusion, but they regained them partly in 1 hour one after another and scored 4 or 5 in the assessment of muscle power next day except for one score of 2; 8 rabbits had their spinal cords impaired severely in the contusion procedure and lost their TMS-MEP too but without recovery, their locomotor capacity outcomes were very poor, 5 of them had no response to transcranial magnetic stimulation next day, and in the other 3 rabbits we only found some polyphase waves with variant latency and lower amplitude which did not resemble common compound muscle action potential (CMAPs) evoked by TMS. CONCLUSIONS: Myogenic TMS-MEPs was very sensitive to the spinal cord injury and should be a valid technique for intraoperative monitoring, and a slight change of them, even if a transient lose, should be unnecessarily related to a severe movement disorder. The warning threshold for a given patient should depend on the malady itself. PMID- 15363297 TI - [Expression and significance of estrogen receptor isoforms in the development of human breast carcinoma]. AB - OBJECTIVE: To investigate the estrogen receptors (ER)alpha and ERbeta expression and their relationship with clinicopathological parameters in human breast carcinoma. METHODS: Samples were obtained from 30 breast carcinoma, reverse transcriptase polymerase chain reaction was used to measure the expression of ERalpha and ERbeta mRNA. RESULTS: ERalpha mRNA level was up-regulated in breast carcinoma tissue compared with adjacent normal tissue (t = 7.399, P < 0.01) while down-regulated in ERbeta. The relative ratio of ERalpha and ERbeta was decreased in normal tissue vs. carcinoma (t = 6.385, P < 0.01), in patients with lymph node metastasis vs. those without lymph node metastasis (t = 2.602, P < 0.05), in late stage carcinoma vs. early stage (t = 3.754, P < 0.05). CONCLUSION: ERalpha and ERbeta play divergent role in the development of human breast carcinoma. PMID- 15363298 TI - [Reversal of multidrug resistance of MCF-7/ADR in nude mice by grape seed polyphenol]. AB - OBJECTIVE: To study the reversing effect of Grape seed polyphenol (GSP) on multidrug resistance of MCF-7/ADR cell in vivo. METHODS: The transplantable breast carcinoma cell line MCF-7/ADR model was established in BALB/C-nu/nu mice by subcutaneous implantation. Flow cytometry (FCM) was used to investigate the changes of Pgp expression and apoptosis rate after different drug treatment. RESULTS: GSP has some effect on inhibition of tumor growth (the rate of inhibition was 18.35%), and combined with adriamycin can significantly inhibit tumor growth in nude mice, 20 mg/kg GSP can effectively reverse the resistance of MCF-7/ADR cells to ADR in vivo, and the rate of inhibition was 54.64%. FCM results showed that the expression of Pgp was significantly decreased (32.03 +/- 2.09) After administration of GSP and ADR, there was distinct difference between it and control (55.13 +/- 2.12). The mean rate of apoptosis was 15.12% +/- 1.04%, and was significantly increased compared with control (9.07% +/- 0.43%), P < 0.05. CONCLUSION: GSP can effectively reversed the resistance of MCF-7/ADR cells in nude mice, the mechanism may be correlated with inhibition of Pgp expression and apoptosis. PMID- 15363299 TI - [Clinical application of mastoscopic axillary sentinel lymph node biopsy]. AB - OBJECTIVE: To evaluate the technique and significance of mastoscopic axillary sentinel lymph node biopsy. METHODS: Sixty-two patients with breast cancer use methylene blue to test axillary sentinel lymph node. Sentinel lymph node was moved with endoscopy, and endoscopic axillary lymph nod dissection was performed. Pathological examination of sentinel lymph node and axillary lymph node was made with HE. To evaluate detection rate and false negative rate in sentinel lymph node. RESULTS: Among the 62 patients, 61 were confirmed by endoscopic axillary sentinel lymph nod biopsy. Detection rate was 98.4%. Thirty-five cases were no metastasis, 27 cases were metastasis, false negative rate was 0. CONCLUSIONS: Mastoscopic axillary sentinel lymph node biopsy has a high detection rate, good efficiency of cosmetic and lower complications. It has higher sensitivity than traditional axillary lymph nod dissection and provide accurate lymph node stages. PMID- 15363300 TI - [Optimal effects of liposome on antisense oligodeoxynucleotides uptake by mouse breast cancer TM40D cells and the effects of antisense oligodeoxynucleotides targeting c-erbB-2 on cell proliferation and apoptosis]. AB - OBJECTIVE: To study the optimal effects of liposome on antisense oligodeoxynucleotides targeting c-erbB-2 uptake by mouse breast cancer TM40D cells and effects of AS-ODNs on cell proliferation and apoptosis. METHODS: Flow cytometric analysis and fluorescence microscope were used to study the transfection ratio of AS-ODNs in mouse breast cancer TM40D cells with/without liposome in 1, 2, 4, 6 hour and the distribution of AS-ODNs in cells. the effects of ODNs on cell proliferation and activation of apoptosis were examined by MTT assay and flow cytometry. RESULTS: Liposome could promote AS-ODNs uptake by mouse breast cancer TM40D cells, flow cytometric analysis revealed that the time of the optimal effects was 4 hours and the percentage of FAM-positive cells reached 72.23%; the ratio maintained at a relatively high level at 6 h. Also liposome facilitated the entrance of AS-ODNs into nucleus. AS-ODNs restrained the proliferation of mouse breast cancer TM40D cells, and restraint rate was 50.24%. Meanwhile AS-ODNs promoted cell apoptosis. Flow cytometry analysis revealed that antisense ODNs increased cell apoptosis by 38.50%, compared with cultured cells group 9.13% and liposome group 9.29%. CONCLUSIONS: Liposome could facilitate AS ODNs to enter cells and their nucleus. AS-ODNs of C-erbB-2 restrained the proliferation of mouse breast cancer TM40D cells and promoted apoptosis. PMID- 15363301 TI - [Application of sentinel lymph node biopsy by subareolar injection of blue dye in the treatment of breast carcinoma]. AB - OBJECTIVE: To probe into the feasibility and accuracy of sentinel lymph node biopsy by subareolar injection of blue dye in the treatment of early-stage breast carcinoma. METHODS: Retrospective analysis of 25 cases of early-stage breast cancer who were carried out subareolar injection of blue dye method for sentinel lymph node biopsy. RESULTS: Of 25 cases, 24 were checked out sentinel lymph nodes. Check-out rate was 96.0%. Totally 62 sentinel lymph nodes were checked out, each patient had 2.5 nodes averagely. There were 5 cases with lymph node metastases in HE stain, and 7 cases with positive nodes in IHC stain. Sensitivity in this group was 87.5%; Accuracy 96.0%; False negative rate 12.5%; False positive rate 0. CONCLUSION: Application of sentinel lymph node biopsy by subareolar injection of blue dye in the treatment of early-stage breast carcinoma can also acquire high check-out rate and high accuracy. PMID- 15363302 TI - [Clinical analysis for 70 sinus of valsalva aneurysm]. AB - OBJECTIVE: To review retrospectively the experience of surgical repair of sinus of valsalva aneurysm (SVA) in 70 patients. METHODS: Between September, 1988 and October, 2003, Seventy patients with SVA underwent surgical repair by the aid of general anesthesia and cardiopulmonary bypass, comprised 1.4% (70/4960) of all open-heart operation. Forty-five were male and 25 female. Age ranged from 3 to 69 years old [mean (29 +/- 15) years]. The aneurysms ruptured into the right ventricle in 46 patients, right atrium in 23 and left ventricle in 1 respectively. The aneurysms originated from right and noncoronary sinus in 61 patients (87%) and 9 patients (13%) respectively. The most common associated cardiovascular lesions were ventricular septal defect (VSD, n = 34) and aortic valve incompetence (n = 21). Repairs were achieved through an incision in right atriotomy, right ventriculotomy or aortotomy only or both aortotomy and right atriotomy (or right ventriculotomy). The defects in the sinus of valsalva was repaired with either direct sutures (n = 43) or a patch (n = 27). The aortic valve was replaced in 6 patients. RESULTS: There were no deaths in early time after repair. Postoperative hospital stay was 8 approximately 33 days [mean +/- standard deviation, (14.3 +/- 6.4) days] before 1997 and 6 approximately 15 days [mean +/- standard deviation, (9.1 +/- 2.6) days] after 1997 respectively. Complications included infection (n = 4), hemorrhage (n = 4), pneumothorax (n = 1), arrhythmia (n = 4) and residual shunt (n = 1) of VSD. Fifty-three (76%) patients (2 months approximately 13 years) were followed-up [mean +/- standard deviation, (6.6 +/- 3.8) years]. All patients survived except that one died of rupture of dissecting aortic aneurysm 7 years after operation. CONCLUSIONS: The ruptured sinus of valsalva aneurysm and unruptured sinus of valsalva aneurysm with ventricle septal defect or(and) aortic valve regurgitation should be repaired surgically as soon as the diagnosis was confirmed. Long-term results are associated with preoperative aortic valve regurgitation. PMID- 15363303 TI - [Total arch replacement combined with stented elephant trunk implantation for Stanford type A aortic dissection]. AB - OBJECTIVE: To improve the long term outcomes of the surgery for Stanford type A aortic dissection, we performed ascending aorta and total aortic arch replacement combined with transaortic stented graft implantation into the descending aorta for acute and chronic type A aortic dissection. METHODS: From April 2003 to March 2004, 40 consecutive patients with acute or chronic Stanford type A aortic dissection underwent this procedure. Right axillary artery cannulation was routinely used for cardiopulmonary bypass and selected cerebral perfusion. The stented elephant trunk was implanted through the aortic arch under hypothermic circulatory arrest. The stented elephant trunk was a 10 cm long self expandable graft. Enhanced electric beam computed tomography (EBCT) was performed in each patient before discharge, three month after surgery, and once each year after discharge to evaluate the postoperative time course of the residual false lumen. RESULTS: Cardiopulmonary bypass time was (166 +/- 38) min, average cross clamp time was (107 +/- 28) min, and average selective cerebral perfusion and lower body arrest time was (30 +/- 15) min. The in-hospital mortality was 5% (2/40). One patient died of multi-organ failure postoperatively and another died of cerebral infarction 2 month after surgery. One suffered from spinal cord injury perioperatively. There was no late death during follow up. CONCLUSION: Ascending aorta and total aortic arch replacement combined with transaortic stented graft implantation into the descending aorta is an effective way in closing the residual false lumen of the descending aorta and might contribute to the better long term outcomes of type A aortic dissection. Our half mortality of 2 patients suffering acute renal failure suggests that this group may be candidates for medical or delayed surgical intervention. Conversely, our 5% mortality rate for those renal intact patients warrant aggressive and expeditious surgical treatment. PMID- 15363304 TI - [The function of the level of serum carcinoembryonic antigen on early recurrence of non-small cell lung cancer]. AB - OBJECTIVE: To explore clinical and pathological factors correlating with early recurrence of resected non small cell lung cancer (NSCLC), and to further understand the function of serum carcinoembryonic antigen (CEA) on NSCLC. METHODS: 93 patients of NSCLC were selected. All of them received resection and were followed up for more than one year. The first time of recurrence was recorded. Logistic univariate and multivariable analysis were used to find the factors that affect the early recurrence of NSLSC, including age, sex, serum CEA level, tumor size, tumor location, tumor differentiation, histological type and clinical staging, and the ability of factors predicting the recurrence were compared by receiver operating characteristic (ROC) curve. RESULTS: Of all the clinical and pathological factors that are correlated with early recurrence of NSCLC, the serum carcinoembryonic antigen (CEA) value, clinical staging, and tumor difference are of statistical significance. The preoperative serum CEA value is the most valuable factor to predict early recurrence of NSCLC (ROC area: 0.843, 95% CI: 0.723 approximately 0.963, P = 0.000). When preoperative serum CEA value > 10 micro g/L, patients of NSCLC will have an early recurrence rate of 88%; and when preoperative serum CEA value 10 micro g/L, even if the lesion is of early stage and well differenced, the general situation of patients should be carefully examined for the prompt and accurate treatment to them and close follow up is needed to treat these patients. PMID- 15363311 TI - [Diagnosis of invasive micropapillary carcinoma]. PMID- 15363313 TI - [Usual hyperplasia, atypical hyperplasia and carcinoma-in-situ of breast: a morphologic study]. AB - OBJECTIVE: To study the morphologic classification of mammary ductal hyperplasia, and its criteria and the significance in distinguishing atypical hyperplasia from carcinoma-in-situ. METHODS: The clinicopathologic features of 300 cases of hyperplasia of breast were reviewed. Whole-organ H&E sections were also available in 86 cases of breast carcinoma. The occurrence of atypical hyperplasia in adjacent breast tissue was assessed. RESULTS: Fibroadenomatoid changes were typically observed in the 21-30 age groups and atypical hyperplasia occurred more frequently in 40-60 age groups. Amongst the hyperplastic cases, cystic diseases of the breast were noted in only 6%. In contrast, fibroadenomatoid changes were more common (25.4%). Atypical ductal hyperplasia occurred in adjacent breast tissue of 65.1% of the carcinoma cases. The incidence was higher (74.9%) if the main lesion was ductal carcinoma-in-situ. CONCLUSIONS: There is a close association between atypical hyperplasia and breast carcinoma. It is prudent to distinguish between usual and atypical hyperplasia. Morphologic differentiation between atypical ductal hyperplasia and ductal carcinoma-in-situ may sometimes be difficult. PMID- 15363312 TI - [Relationship between expression of cell adhesion molecules and metastatic potential in invasive micropapillary carcinoma of breast]. AB - OBJECTIVE: To investigate the expression of cell adhesion molecules and Their significance in invasive micropapillary carcinoma (IMPC) of the breast. METHODS: Immunohistochemical study for E-cadherin was performed on 64 cases of IMPC and 57 cases of invasive ductal carcinoma (IDC). RESULTS: E-cadherin was mainly expressed on the cell membrane of tumor cells. The expression of E-cadherin in IMPC (85.9%, 55/64) was significantly higher than that in IDC (43.9%, 25/57). E cadherin expressed in the intercellular contact surface of IMPC cells. In contrast, it was weakly positive/not expressed on the outer membranous surface of the tumor clusters in IMPC. The rate of lymph node metastasis in IMPC (85.9%, 55/64) was significantly higher than that in IDC (52.6%, 30/57), the rate of alpha-catenin and beta-catenin coexpression in IMPC (45.1%, 26/51) with lymph node metastasis and E-cadherin normal expression was also significantly higher than that in IDC (15.4%, 2/13). CONCLUSION: Weak cell adhesion molecule expression on the outer surface of IMPC cell clusters, in contrast to strong cohesion in intercellular contact surface, may help to explain the high metastatic potential of this type of breast cancer. PMID- 15363314 TI - [Role of cytokeratin expression in differential diagnosis of intraductal proliferative lesions of breast]. AB - OBJECTIVE: To evaluate the expression of cytokeratins in intraductal proliferative lesions of breast, including usual ductal hyperplasia (UDH), atypical ductal hyperplasia (ADH), ductal carcinoma-in-situ (DCIS) and its role in differential diagnosis. METHODS: Ninety two cases of paraffin-embedded lesional breast tissue, 30 cases of frozen samples, cell cultures of hyperplastic ductal cells and 2 invasive ductal carcinoma cell lines (T47D and MCF-7) were used for this study. Immunohistochemistry was performed using EnVision method for 34betaE12, CK8 and CK14. RESULTS: The percentage of 34betaE12-positivity in paraffin-embedded samples of UDH, ADH, DCIS and invasive ductal carcinoma (IDC) was found to be 95.2%, 33.3%, 19.2% and 12.5% respectively. In frozen tissues, all UDH cases and 55% of IDC cases expressed 34betaE12. The primary UDH cell cultures and T47D cell line were also 34betaE12-positive, whereas MCF7 cell line showed negative staining. The expression rate of CK8 and CK14 in UDH was also different from that in ADH and DCIS. CONCLUSIONS: 34betaE12 can be useful in differential diagnosis of intraductal proliferative lesions of the breast. However application of this cytokeratin stain in intraoperative frozen sections is relatively limited. The expression patterns of CK8 and CK14 are also helpful in the differential diagnosis of similar lesions. PMID- 15363315 TI - [Expression of PH20 in primary and metastatic breast cancer and its pathological significance]. AB - OBJECTIVE: To evaluate the expression pattern of PH20 in primary and metastatic breast cancer and its relationship to tumor metastatic potential. METHODS: Anti PH20 antibody was synthesized by injection of conjugated human PH20 peptides into rabbits. Immunohistochemical study was performed on 53 cases of human breast cancer. Western blot was used to detect PH20 expression in 5 cases of breast cancer with available fresh tissue. Two oligonucleotide probes were prepared for in-situ hybridization using breast tissue microarray. RESULTS: Normal breast tissue did not express PH20 (0/3), while 58.4% (31/53) of breast cancer cases did. The highest expression rate was found in metastatic foci in regional lymph nodes (83.3%), followed by primary breast cancer tissue in cases with lymph node secondaries (70.8%). The breast cancer cases with no any metastasis had an expression rate of 48.2%. The immunohistochemical staining results were further confirmed by Western blotting. In-situ hybridization showed PH20 RNA in 75% of the breast cancer tissue (21/28). Two of the 17 cases of normal breast tissue showed weak expression in some ductolobular units. CONCLUSIONS: The expression of PH20 has a positive correlation with metastatic potential in breast cancer. It is possible that PH20 may play an important role in the invasive growth and metastasis of breast cancer cells, via mechanisms such as digestion of surrounding stromal tissue and release of FGF-2. PMID- 15363316 TI - [Evaluation of p63 expression in lung cancer by use of complementary DNA and tissue microarray]. AB - OBJECTIVE: To study p63 expression at mRNA transcript and protein levels in human lung cancers, including squamous cell lung carcinoma (SCC), adenocarcinoma, large cell lung carcinoma (LCLC) and small cell lung carcinoma (SCLC), or the corresponding metastatic foci. The relationship of p63 expression and alterations in p63 locus at chromosomal 3q27-29 was also determined. METHODS: p63 gene expression in 72 cases of SCC, adenocarcinoma, LCLC and SCLC was analyzed by cDNA microarray technology. Tissue microarray of specimens from 150 cases of primary lung cancer was prepared for immunohistochemical study for p63 protein. Possible chromosomal alterations at the p63 locus in 70 cases of primary lung cancer were studied by comparative genomic hybridization (CGH) technology. RESULTS: p63 mRNA transcript expression was significantly increased by more than 10-fold in SCC, as compared with that in other histologic subtypes including adenocarcinoma, LCLC and SCLC. p63 mRNA expression in metastatic foci was also remarkably higher than that in their primary tumors (P < 0.001). Immunostaining showed that p63 protein expression was observed in 94.64% of SCC, whereas only one lung adenocarcinoma (1.79%) was positive. Immunopositivity was also demonstrated in 2 of the 4 LCLC cases studied. None of the SCLC cases was positive. There was a statistically significant difference in p63 expression between pT1 and pT2 tumors (P < 0.05). The CGH results showed that overrepresentation of p63 locus at chromosomal 3q27 29 was a typical finding in SCC. p63 immunopositivity also correlated significantly with pronounced gains of p63 locus at chromosomal 3q27-q29 (P < 0.000 1), suggesting that strong expression of p63 in lung SCC was associated with increased gene amplification. CONCLUSION: p63 may play a role in oncogenesis of human lung squamous cell carcinoma and development of metastasis. PMID- 15363317 TI - [Detection of EWS-FLI1 fusion transcript in Ewing's sarcoma/peripheral primitive neuroectodermal tumors by one-step RT-PCR using paraffin-embedded tissues]. AB - OBJECTIVE: To investigate the expression of EWS-FLI1/ERG fusion transcript resulting from t(11;12)(q24;12) in paraffin-embedded tissues and its diagnostic implication for Ewing's sarcoma/peripheral primitive neuroectodermal tumors (ES/pPNET). METHODS: One-step reverse transcriptase-polymerase chain reaction (RT PCR) was employed to detect a characteristic EWS-FLI1/ERG fusion transcript in 25 cases of ES/pPNET and 15 cases of other small round cell tumors (including 8 cases of rhabdomyosarcoma, 4 cases of synovial sarcoma, 2 cases of neuroblastoma and 1 case of lymphoma) using formalin-fixed and paraffin-embedded tissues. RESULTS: EWS-FLI1/ERG fusion transcript was detected in 20 of the 25 ES/pPNET cases (80%). The 15 non-ES/pPNET control cases were negative for EWS-FLI1/ERG fusion transcript. CONCLUSIONS: Detection of EWS-FLI1/ERG fusion transcript is a reliable index for molecular diagnosis of ES/pPNET. One-step RT-PCR is a practical method for such analysis in routine paraffin-embedded tumor tissues. PMID- 15363318 TI - [Study on MYCN gene amplification and CD44 expression in neuroblastoma]. AB - OBJECTIVE: To assess the relationship between MYCN amplification in neuroblastoma (NB), tumor stage and prognosis; and to evaluate the usefulness of CD44 in predicting prognosis of NB. METHODS: Differential polymerase chain reaction (D PCR) with serial dilution assay was used to quantify the MYCN gene copy number in 33 paraffin-embedded tissue samples of NB. All the samples were also studied by immunohistochemistry for CD44. The results were correlated with various prognostic factors of NB, including patient age, tumor stage, pathologic type and MYCN gene amplification. RESULTS: MYCN amplification was identified in 10 of the 33 samples studied (30.3%), which all were in high clinical stage (stage III or IV) and occurred in patients older than 1 year of age. MYCN amplification also significantly correlated with poor clinical outcome (P < 0.01). CD44 was positive in 21 cases and often occurred in patients below 1 year of age, in low clinical stage, with favorable histology and without MYCN gene amplification. The two-year survival rate of CD44-positive group (57.1%) was higher than that of CD44 negative group (8.3%, P < 0.01). Stronger CD44 expression was also associated with better prognosis (P < 0.01). CONCLUSIONS: MYCN gene amplification is significantly associated with advanced tumor stage and poor prognosis in patients with NB. CD44 expression is a reliable marker for better prognosis and is complementary to MYCN amplification assay. D-PCR with serial dilution assay is also suitable for clinical use in quantifying MYCN copy number in NB. PMID- 15363319 TI - [Sclerosing epithelioid fibrosarcoma: a clinicopathologic study of eight cases]. AB - OBJECTIVE: To study the clinicopathologic features, immunophenotype and differential diagnosis of sclerosing epithelioid fibrosarcoma (SEF). METHODS: Eight cases of SEF were investigated by light microscopy and immunohistochemistry. RESULTS: There were five males and three females. Clinically, most patients presented as a slowly growing mass. Six tumors were located in the extremities or limb girdles, and two in the trunk. Grossly, most lesions were relatively well-circumscribed with a nodular or lobulated appearance. They ranged from 2.0 to 10.5 cm in size (mean 6 cm). On sectioning, they had a gray-whitish cut surface and were firm in consistency. Microscopically, the tumors were composed of uniformly round or polygonal epithelioid cells with clear to eosinophilic cytoplasm. The tumor cells were arranged predominantly in single strands or cords and embedded in a heavily hyalinized matrix. In some areas, nests, sheets, acini or alveolar structures were also noted. Nuclei atypia and brisk mitotic activity was not evident. The mitotic count measured less than 1 per 10 high power fields. However in two cases, focal areas exhibited increased cellularity, nuclei atypia and higher mitotic activity, resembling conventional fibrosarcoma. Immunohistochemically, the tumor cells showed diffuse and strong positivity for vimentin and focal or weak positivity for EMA. There was no expression for AE1/AE3, S-100 protein, HMB45, alpha-SMA, MSA, desmin, CD34, bcl-2, CD30 and LCA. Follow-up information in six patients revealed local recurrence in 3 cases and lung metastasis in 1 case. CONCLUSIONS: SEF is a rare variant of fibrosarcoma. Despite the relatively bland appearance and low mitotic activity, the tumor is capable of local recurrence and distant metastasis. Thus, it should be considered and treated as a low to intermediate grade sarcoma. SEF needs to be differentiated from a variety of benign or malignant tumors exhibiting epithelioid features and sclerotic stromal response. PMID- 15363320 TI - [Application of immunohistochemistry-laser microdissection-PCR technique in detecting p53 gene mutation in paraffin sections of gastric cancer]. AB - OBJECTIVE: To evaluate the application of immunohistochemistry-laser microdissection-polymerase chain reaction (IHC-LMD-PCR) technique in detecting p53 gene mutations using paraffin sections of advanced gastric cancers. METHODS: The expression of p53 protein in 41 paraffin-embedded advanced gastric cancer samples was assessed by immunohistochemistry. The p53-positive or negative carcinoma cells and normal foveolar cells far from the main tumor were then isolated by LMD. The DNA was extracted and p53 gene at exon 5 - 8 amplified by PCR. The products were then analyzed by single-strand conformation polymorphism (SSCP) and automated sequencing. RESULTS: p53 gene was successfully amplified in all the 41 specimens. Overexpression of p53 protein was noted in 11 cases. Mutations of p53 gene were found in 15 cases. Eight of the 11 p53-overexpressed cases had p53 gene mutations. On the other hand, p53 gene mutations were found in only 7 of the 30 p53 protein-negative cases. The presence of p53 gene mutations significantly correlated with p53 protein overexpression (P = 0.004). CONCLUSIONS: IHC-LMD-PCR technique can be successfully applied in paraffin sections of gastric cancers for the detection of p53 gene mutations. The results correlate well with overexpression of p53 protein. PMID- 15363321 TI - [Loss of heterozygosity in esophageal squamous cell carcinoma and its precursor lesion]. AB - OBJECTIVE: To detect the loss of heterozygosity (LOH) in esophageal squamous cell carcinoma and adjacent high-grade squamous dysplasia, and to evaluate possible tumor suppressor genes in the development and progression of invasive malignancy. METHODS: LOH was detected in normal esophageal mucosa, high grade squamous dysplasia and esophageal squamous cell carcinoma using microdissection and polymerase chain reaction technology. The changes of LOH at seven microsatellite markers and the relationship between LOH rate and clinicopathologic parameters were analyzed. RESULTS: In high grade squamous dysplasia, LOH was detected at D13S802 (40%), D13S267 (32%), D13S221 (31%), D9S942 (30%), D17S520 (24%) and D9S171 (33%). However, D17S1798 LOH was not detected. In invasive squamous cell carcinoma, LOH was detected as follows: D13S267 (71%), D13S802 (58%), D17S520 (55%), D13S221 (45%), D9S942 (43%), D9S171 (33%) and D17S1798 (11%). The frequency of LOH in the seven microsatellite markers, the pathologic grade, clinical stage and occurrence of lymph node metastasis did not show any statistically significant correlation (P > 0.05). CONCLUSIONS: The progression from normal squamous epithelium to high grade squamous dysplasia and subsequently to invasive squamous cell carcinoma of the esophagus was associated with accumulation of genetic errors. Possible tumor suppressor genes related to the development of esophageal squamous cell carcinoma may exist near D13S802 (13q12.12). Possible tumor suppressor genes near D13S267 (13q13.1), D17S1798 (17p13.3) and D17S520 (17p13.1) may be related to the progression of esophageal squamous cell carcinoma. PMID- 15363322 TI - [Expression and significance of DARPP-32 in gastric carcinoma]. AB - OBJECTIVE: To investigate the significance of DARPP-32 protein expression in gastric carcinoma tissue and cell lines. METHODS: The expression of DARPP-32 protein in normal gastric mucosa and gastric carcinoma tissue was evaluated by immunohistochemical staining using streptavidin-biotin complex technique. The expression in gastric carcinoma tissue and cell lines was evaluated by Western blotting. RESULTS: The expression rate of DARPP-32 protein in gastric adenocarcinoma tissue (92.7%) was significantly higher than that in normal gastric mucosa (52.6%, P < 0.05). There was no significant association between DARPP-32 protein expression and degree of tumor differentiation, local invasion and distant metastasis. As compared with adjacent non-carcinomatous gastric mucosa, both DARPP-32 and its truncated isoform t-DARPP were overexpressed in gastric adenocarcinoma tissue (t = 2.45, P = 0.015); and t-DARPP overexpression was more frequently seen. Expression of DARPP-32 and t-DARPP could also be detected in human gastric cancer cell lines. The expression of DARPP-32 protein was obviously reduced in SGC7901 drug-resistant cell strains. CONCLUSIONS: DARPP 32 is overexpressed in gastric carcinoma. It may play an important role in gastric carcinogenesis. The underlying signal pathways in neoplastic gastric epithelium may also be related to the multi-drug resistance property of gastric cancer cells. PMID- 15363323 TI - [Pathological changes of lungs in patients with severity acute respiratory syndrome]. AB - OBJECTIVE: To evaluate the progression in morphologic changes of lungs in SARS patients. METHODS: Four cases of SARS with lung tissue samples available (including one for ultrastructural examination) were enrolled into the study. Histochemical study for VG, Masson, reticulin, orcein, PAS, sirius red stains and immunohistochemical study for vimentin, desmin, smooth muscle actin, HHF-35, CD34, F8, collagen types I and III were also performed. RESULTS: According to the morphologic changes, lung lesions in SARS were subcategorized into 3 phases: acute exudative inflammation, fibrous proliferation and the final fibrotic stage. Two cases belonged to the acute exudative phase, in which the course was less than 20 days. The principal lesions consisted of acute alveolar exudative inflammation, hyperplasia of alveolar epithelium, necrosis, alveolar hyaline membrane formation, alveolar desquamation and focal fibroplasia. The acute exudative protein was PAS-positive. There was an increase in reticulin fiber formation. The reactive fibroblasts were highlighted by desmin and vimentin. One case belonged to the fibroproliferative stage, in which the course was around 25 days. Major lesions included proliferative interstitial pneumonia with early pulmonary fibrosis. There was also evidence of organizing pneumonia, with an increase in reticulin fiber formation, which had a glomeruloid appearance on special stain. The mesenchymal cells showed either myofibroblastic (which expressed desmin, HHF-35, smooth muscle actin and vimentin) or fibroblastic (which expressed vimentin only) differentiation. Fibroelastosis and fibroplasia was also noted. The remaining case belonged to the fibrotic stage, in which the course was around 75 days. The main features included diffuse fibrosis and honeycomb change, which were highlighted by sirius red stain. Immunohistochemistry showed mainly types I and IV collagen fibers. In all lesions, there was also an increase of number of CD68-positive macrophages. CONCLUSIONS: The morphologic progression in lungs of SARS patients is characterized by the development of increased fibrosis. The primitive mesenchymal cells, hyperplastic alveolar epithelial cells and macrophages play an important role in the pathogenesis. PMID- 15363324 TI - [Overexpression of HER2/neu downregulates wild p53 protein expression via PI3K and Ras/Raf/MEK/ERK pathways in human breast cancer cells]. AB - OBJECTIVE: To evaluate the effect of HER2/neu overexpression on wild p53 protein expression and to delineate the related signal pathways. METHODS: Lipofectin method was used to transfer HER2/neu into human breast tumor cell line MCF7. Overexpression of HER2/neu was then determined by Western blot. Western blot was also used to detect the quantity of p53, Akt, p-Akt, p-Raf, p-MEK, p-ERK protein. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to detect p53 mRNA expression. PI3K pathway inhibitor LY294002 and MEK inhibitor U0126 were used to block the two pathways. The subsequent effect on p53 protein expression was then determined. RESULTS: HER2/neu-overexpressed MCF7 clone (MCF7-neu3) was successfully established, in which the amount of HER2/neu protein was 13 times more than that in parental MCF7 cells. The amount of p53 protein in MCF7-neu3 was 40% of that in parental MCF7 cells (P < 0.01), while there was no difference on p53 mRNA level. There were 2.5, 2.0, 1.6 and 1.6 fold increase in the amount of p-Akt, p-Raf, p-MEK, p-ERK protein respectively in MCF7 neu3 to that in parental MCF7 cells (P < 0.01). When treated with LY294002 or U0126 for 24 hours, the amount of wild p53 protein in MCF7-neu3 cells was 1.7 or 1.5 times higher than those in DMSO treated cells. There were 4.7 or 5.3 times increase in the p53 protein when MCF7-neu3 cells were treated with LY294002 or U0126 for 48 hours (P < 0.01). Similar results were not seen in MDA-MB-453 cells which contained mutant p53. CONCLUSIONS: HER2/neu overexpression can activate PI3K and Ras/Raf/MEK/ERK pathways, resulting in reduction of wild p53 protein expression. This may be the molecular mechanism responsible for the poor prognosis and therapeutic non-responsiveness in HER2/neu-overexpressed breast cancer patients. PMID- 15363325 TI - [The role of Egr-1 and NF-kappaB in the pathogenesis of silicosis: an in-vitro study]. AB - OBJECTIVE: To study the correlation between the expression of Egr-1 and NF-kappaB and the up-regulation of TNF-alpha and TGF-beta1 in macrophages after stimulation by silica in-vitro. METHODS: Macrophages were treated with antibodies against Egr 1 and NF-kappaB and antisense oligonucleotides. The level of TNF-alpha protein in the cell supernatant was then measured using enzyme-linked immunoadsorbent assay (ELISA). The expression of TGF-beta1 protein was detected by immunocytochemistry. The expression of TNF-alpha and TGF-beta1 mRNAs was also monitored by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: Compared with silica stimulated macrophages untreated with antibodies, the cells treated with 10 micro g/ml of Egr-1 or NF-kappaB antibodies were associated with reduced expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05). Compared with silica stimulated untransfected group, the antisense group was associated with obvious reduction in the expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05). CONCLUSION: The expression of TNF-alpha and TGF-beta1 mRNAs and proteins are associated with activation of Egr-1 and NF-kappaB in macrophages, after stimulation by silica. It is possible that the corresponding antibodies and antisense oligonucleotides may become a potential therapeutic tool in the management of silicosis in the future. PMID- 15363330 TI - [Advances of RET oncogene activation in papillary thyroid carcinoma]. PMID- 15363342 TI - [Analysis on incidence for cervical cancer in Qidong during 1972 - 2000]. AB - OBJECTIVE: To investigate the trends of incidence for cervical cancer in the years of 1973 - 2000, in Qidong, a city in south China, with a population of 1.16 M. METHODS: Crude incidence rate of cervical cancer was calculated by using incident case data from Qidong Cancer Registry, which was founded in 1972, and age-standardized rates were computed according to China age structure of 1964 (CASR), and world age structure of 1960 (WASR). The epidemiological features and secular change trends of the cancer were described. RESULTS: Six hundred and thirty-one new cases of cervical cancer were reported in Qidong during the years of 1973 to 2000, ranking the eighth and accounting for 3.25% among all sites of cancers in women. The crude annual incidence rate during the period was 3.96 per 100,000 on average, China age-standardized rate (CASR) was 2.67 per 100,000, and world age-standardized rate (WASR), 3.66 per 100,000. The incidence of cervical cancer was 2.97 per 100,000 for the last 14 years, and 5.01 per 100,000 for the first 14 years, showing a decreasing trend (P < 0.01). Age-specific rates showed that the incidents among those aged over 55 in the recent 14 years were markedly lower than that in the first 14 year period (P < 0.01). CONCLUSIONS: Qidong is a relatively low incidence area of cervical cancer, with a decreasing trend among aged women, according to population-based cancer registration data. This incidence data on cervical cancer could be used as a base for strategic treatment and prevention of the disease. PMID- 15363343 TI - [Study on combined transvaginal hydrolaparoscopy and hysteroscopy in patients with infertility]. AB - OBJECTIVE: To evaluate the advantages of combined transvaginal hydrolaparoscopy (THL) and hysteroscopy in patients with infertility. METHODS: Twenty-three primary infertile patients (primary group) and 23 secondary infertile patients (secondary group) were received THL (using a 3-mm scope with a 0 degrees lens) and hysteroscopy. The fallopian tube patency, pelvic pathology, operative time, discharge time, healing time of vaginal wound, success rate of insertion into the pouch of Douglas and intra- and postoperative complications were observed. RESULTS: Primary group had more tube patency cases (P < 0.05) and fewer pelvic adhesion cases (P < 0.05) when compared with secondary group. Only 15.2% patients needed conventional laparoscopy postoperatively. The time of THL, overall time of operation, discharge time, and healing time of vaginal wound were respectively (10.5 +/- 1.7) min, (32.2 +/- 4.6) min, (87.3 +/- 12.5) min, and (4.5 +/- 0.5) days. No complications occurred. CONCLUSIONS: THL is accurate, minimally invasive, safe, economical, and does not require hospitalization. It could serve as a first-line procedure in early stages of infertility investigation when combined with hysteroscopy. PMID- 15363344 TI - [Study on the time selection of pregnancy and delivery in women with systemic lupus erythematosus]. AB - OBJECTIVE: To analyze the appropriate time selection of pregnancy and delivery in women with systemic lupus erythematosus (SLE). METHODS: Twenty-nine pregnancies in women with SLE in our hospital from 1998 to 2003 were retrospectively analyzed regarding the selection of appropriate time of pregnancy and delivery. RESULTS: All patients did not take any cytotoxic medicine for at least 6 months before pregnancy. Twenty-three conceptions occurred when SLE was inactive for at least 1 year. Two conceptions occurred when SLE was active without doctors' agreement. SLE was diagnosed during pregnancy in the remaining 4 cases. The condition of all patients fluctuated and the gestational time at delivery ranged from 30 to 38 weeks after we modified the doses of glucocorticoid (prednisone). Among totally 29 living neonates, eight were premature neonates, three were FGR and one had serious congenital heart disease. Two neonates died of complications in early stage of neonatal period. None of the 29 neonates from all patients had neonatal lupus. CONCLUSION: Pregnancy safety will be improved obviously if the condition of SLE is controlled and the patients are given reasonable doses of glucocorticoid and intensive monitoring. If pharmacotherapy does not work well and the condition threatens the safety of mother and fetus, or the fetus has matured, termination of pregnancy should be done on time, which reduces maternal complications and improves the perinatal mortality rate. The gestational time should be 34 to 38 weeks. PMID- 15363346 TI - [Clinical analysis of anomalies of female reproductive system associated with urinary system]. AB - OBJECTIVE: To determine the association of anomalies of female reproductive system with that of urinary system. METHODS: Two hundred and eighty-nine cases, including 59 with congenital vaginal inclined septum syndrome, 135 with septate uterus, 4 with unicornuate uterus, 13 with rudimentary horn of uterus, 58 with congenital absence of the uterus and vagina, 14 with imperforated hymen, and 6 with atresia of vagina, treated during the past 4 years, were analysed retrospectively. RESULTS: Anomalies of urinary system were more frequently found in patients with anisomerous anomalies of reproductive system (85.53%, 65/76 cases). In 59 cases of congenital vaginal inclined septum syndrome, 43 had ipsilateral renal agenesis, 2 had one single pelvic kidney, 1 had bilateral horseshoe kidneys, 1 had left-side horseshoe kidney, 1 had one pair of renal pelvis and renal calices with ureters incorporated at middle and inferior segments on both sides. Ipsilateral renal agenesis was also seen in patients with rudimentary horn of uterus (14/14 cases) and unicornuate uterus (4/4 cases). However, there was no urinary system malformation seen in septate uterus patients (0/135 cases), imperforated hymen (0/16) and atresia of vagina (0/4 cases). Only 1 case of congenital absence of the uterus and vagina associated with ipsilateral renal agenesis. CONCLUSION: Anomalies of reproductive ducts associated with renal malformations, especially anisomerous anomalies of reproductive system are not unusual. Early recognition and diagnosis is very important to prevention and treatment of such diseases. PMID- 15363348 TI - [Study of morphological changes in levator ani muscle of patients with stress urinary incontinence or pelvic organ prolapse]. AB - OBJECTIVE: To investigate the morphological characteristics of levator ani muscle in patients with stress urinary incontinence (SUI) or pelvic organ prolapse (POP) and to explore whether the alterations could contribute to pathogenesis of the diseases. METHODS: Biopsy specimens of levator ani muscle were obtained from 15 patients with SUI, 19 patients with POP and 3 asymptomatic controls with rectal cancer during operation. The structure of levator ani muscle was examined with routine histological techniques: HE staining, modified Gomori trichrome staining, NSE staining, ACP staining, and adenosine triphosphatase (ATPase) staining. RESULTS: There was no significant difference in age, parity, menopausal time, disease severity and leak point pressure between SUI patients with or without muscle fibers (P > 0.05). The muscular fiber density of levator ani muscles in SUI and POP groups was decreased, fibers were arranged in disorder and separated by large quantities of dense connective tissues with infiltrating inflammatory cells. The muscle fiber fascicles showed obvious grouped denervative atrophy, fiber type grouping and angular in shape. And also there was myopathic degeneration such as centrally located nuclei, peripheral phagocytosis and vacuolated necrosis. CONCLUSIONS: There are both neurogenic and myopathic alterations in levator ani muscle's structure in patients with SUI and POP. The presence of both acute and chronic abnormalities indicates that the weakness of pelvic floor is a consequence of prolonged denervation. PMID- 15363349 TI - [Outcome of follicular aspiration of selection phase in patients with polycystic ovary syndrome undergoing controlled ovarian hyperstimulation]. AB - OBJECTIVE: To assess the effectiveness of follicular aspiration in selection phase of follicle growth of the patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization-embryo transfer (IVF-ET). METHODS: Thirteen infertile patients with PCOS who previously failed IVF-ET associated with ovarian hyperstimulation syndrome (OHSS), served as control group, were asked to undergo new IVF cycles (follicular aspiration group) with ultrasound-guided transvaginal follicular aspirations in the selection phase (diameter 9 - 10 mm) of follicle growth, with a total of 13 cycles both. No more than 10 follicles were selected. Clinical and endocrinological parameters were monitored. RESULTS: In follicular aspiration group, the average number of follicle >or= 12 mm in diameter and serum estradiol level were 16 +/- 4 and (9899 +/- 1431) pmol/L respectively, which were significantly lower than 29 +/- 8 (P < 0.001) and (15,544 +/- 1767) pmol/L (P < 0.001) in control group. No moderate-severe OHSS occurred in follicular aspiration group, while 9 out of 13 (69%) occurred in control group. Oocyte retrieval rate, maturation rate, fertilization rate and cleavage rate in follicular aspiration group were greatly higher than those in control group. CONCLUSIONS: Follicular aspiration in selection phase of follicle growth associates with reduced dominant follicle numbers and estradiol level on the day of hCG injection and moderate-severe OHSS incidence in patients with PCOS undergoing IVF-ET. PMID- 15363350 TI - [Cryopreservation of mouse morula and blastocysts by vitrification]. AB - OBJECTIVE: To compare the survival and developmental potential of mouse morula, early blastocysts and blastocysts cryopreserved by vitrification. METHODS: One hundred and forty-two mouse morula, 135 early blastocysts and 148 blastocysts were cryopreserved by 6 mol/L ethylene glycol and 1 mol/L sucrose vitrification solutions. The survival rates and blastocysts hatching rates after thawing were observed. RESULTS: The survival rates of morula, early blastocysts and blastocysts groups were 88.0%, 73.3%, and 60.1% respectively. The blastocyst hatching rates were 73.9%, 61.5%, and 49.3% respectively. Both the survival rates and blastocyst hatching rates in morula group were higher than those in early blastocysts group (P < 0.05), and they were higher in early blastocysts group than in blastocysts group (P < 0.05). CONCLUSIONS: In these three embryo stages, the survival and developmental potential of mouse morula cryopreserved by vitrification are better than those of early blastocysts and blastocysts. Morula is likely the most feasible stage for embryo vitrification. PMID- 15363351 TI - [Relationship of cyclooxygenase 2 expression and chemotherapy response and prognosis in human ovarian carcinoma]. AB - OBJECTIVE: To investigate the relationship between expression of cyclooxygenase 2 (COX-2) and chemotherapy response and prognosis of patients with ovarian carcinoma. METHODS: The expression of COX-2 was detected by immunohistochemistry in ovarian carcinoma tissues. Correlations between COX-2 and clinicopathological markers, therapeutic effect and prognosis of the patients with ovarian carcinoma were analyzed by chi(2) test, Kplan-Meire and Cox model. RESULTS: COX-2 positive rate of 70 ovarian carcinomas was 60%. Expression of COX-2 was correlated with lymph nodes metastasis only (P = 0.001). There was no correlation between COX-2 and pathohistological types, clinical stage, pathohistological grade, age at diagnosis (P = 0.397, 0.094, 0.275, 0.394). In patients whose serum CA(125) level did not drop to normal after two courses of chemotherapy, the COX-2 positive rate (76%) was significantly higher than in those whose serum CA(125) level decreased obviously (51%, P = 0.042). The positive rate was also higher in patients with recurrent diseases (67%) than in those without recurrence (46%, P = 0.062). In univariate survival analysis, the expression of COX-2 was associated with a significantly reduced median survival time (P = 0.029). In multivariate survival analysis, COX-2 was an independent prognostic factor for poor survival (P = 0.036). CONCLUSION: COX-2 expression is correlated with poor chemotherapy response and prognostic outcome. It is an independent prognostic factor of ovarian carcinoma. PMID- 15363352 TI - [Influence of nuclear factor-kappaB decoy transfection on nuclear factor-kappaB activity and vascular endothelial growth factor/urokinase-type plasminogen activator/intercellular cell adhesion molecule-1 level of SKOV3 cells]. AB - OBJECTIVE: To investigate the effect of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotides (ODN) on NF-kappaB activation and expressions of vascular endothelial growth factor (VEGF), urokinase-type plasminogen activator (uPA), intercellular cell adhesion molecule-1 (ICAM-1) of SKOV3 cell. METHODS: After transfected with NF-kappaB decoy ODN, SKOV3 cells were stimulated by IL-1beta and cultured for 6, 12, 24, and 48 h, respectively. NF-kappaB DNA binding activity was measured by gel mobility shift assay and mRNA levels of ICAM-1, uPA, VEGF were analyzed by RT-PCR and VEGF level in supernatant was detected by enzyme linked immunosorbent assay (ELISA). RESULTS: (1) SKOV3 cells expressed constitutive NF-kappaB DNA binding activity and stimulation of IL-1beta resulted in a significant increase of the binding activity. NF-kappaB decoy ODN transfection significantly decreased constitutive level and elevated level of NF kappaB DNA binding activity by IL-1beta at 6, 12, 24, and 48 h in vitro (P < 0.05). (2) The constitutive and activated mRNA levels of uPA, VEGF, ICAM-1 and VEGF levels in the supernatant were significantly inhibited by NF-kappaB decoy ODN transfection especially at 6 h after transfection in vitro. CONCLUSIONS: Both NF-kappaB DNA binding activity and expressions of uPA, VEGF, and ICAM-1 at 48 h were upregulated by IL-1beta, and inhibited by transfection with NF-kappaB decoy ODN in SKOV3 cell line. NF-kappaB decoy ODN transfection shows promise as a novel molecular approach for ovarian cancer. PMID- 15363353 TI - [Expression of pituitary tumor-transforming gene in endometrial carcinoma]. AB - OBJECTIVE: To study the expression of pituitary tumor-transforming gene (PTTG) and its relationship with the expression of basic fibroblast growth factor (bFGF) protein and microvessel density (MVD) in endometrial carcinoma. METHODS: Expressions of PTTG mRNA and protein were assessed by semi-quantitive RT-PCR and immunohistochemistry methods respectively in 50 cases of endometrial carcinomas, 15 cases of hyperplasia endometria and 12 cases of normal endometrial tissue. Expressions of bFGF protein were detected by immunohistochemistry. Microvessels were highlighted by staining endothelial cells with CD(34) antigen, and MVDs were counted. RESULTS: The expression rate and average quantity of PTTG mRNA were detected in a significantly greater proportion endometrial carcinomas (96%, 0.84 +/- 0.08) than in hyperplasia endometria (60%, 0.78 +/- 0.06) and normal endometrial tissue (33%, 0.48 +/- 0.12, P < 0.01, respectively). The expression of PTTG protein in endometrial carcinomas (70%) was significantly higher than in hyperplasia endometria (40%) and normal endometrial tissue (17%, P < 0.01). The expression of PTTG was related to surgical-pathological stage, myometrial infiltration depth, lymphatic metastasis and pathological subtype (P < 0.05, respectively), but was irrelevant to patients' age and pathological grade (P > 0.05, respectively). The average quantity of PTTG mRNA and expression rate of PTTG protein in tissues with bFGF protein coexpression (0.86 +/- 0.07, 87%) were higher than in those without bFGF protein coexpression (0.80 +/- 0.06, 42%, P < 0.01, respectively). The MVD in tissues with PTTG protein expression (62 +/- 18) was higher than in those without PTTG protein expression (51 +/- 12, P < 0.05). CONCLUSIONS: PTTG may play an important role in carcinogenesis and development of endometrial carcinoma. PTTG induces an angiogenesis through bFGF which is a key determinant step in tumor progression and metastatic spread. PMID- 15363354 TI - [Effects of nitric oxide synthase inhibitors on cerebral mitochondrial structure and function in fetal rats with intrauterine distress]. AB - OBJECTIVE: To evaluate the effects of different types of nitric oxide synthase (NOS) inhibitors on cerebral mitochondrial structure and function in fetal rats with intrauterine distress. METHODS: Rats were divided into control group, acute ischemia group, treatment group 1 injection of [N omega-nitro-L-arginine (L-NNA) 4 mg/kg into pregnant rats' abdomen before ischemia], reperfusion group, treatment group 2 [injection of L-NNA 4 mg/kg into pregnant rats' abdomen before ischemia followed by injection of aminoguanidine (AG) 500 mg/kg before operation]. Changes of mitochondrial structure were observed by transmission electron microscopy and expression of neuronal nitric oxide synthase (nNOS) mRNA(A) through RT-PCR. Inducible nitric oxide synthase (iNOS) activity and mitochondrial Na(+)K(+)-ATPase and cytochrome oxidase activity were measured. RESULTS: (1) The A of NOS (5 min, 15 min) in acute ischemia group was higher than that of treatment group 1 (P < 0.05). There was no difference between the A of nNOS (30 min) in two groups (P > 0.05). But the A of nNOS in two groups was higher than that in control group (P < 0.05). (2) iNOS activities in reperfusion group were all higher than those in treatment group 2. Both of those in two groups were higher than that in control group (P < 0.05). (3) Mitochondrial Rsv (5 min, 15 min) in acute ischemia group was smaller than those of treatment group 1 (P < 0.05). There was no difference between mitochondrial Rsv (30 min) in two groups (P > 0.05). Mitochondrial Rsv in reperfusion group was all smaller than those in treatment group 2. And mitochondrial Rsv in all the groups was smaller than that in control group (P < 0.05). (4) Na(+)K(+)-ATPase activity in treatment group 2 was higher than those in reperfusion group (P < 0.05). Na(+)K(+)-ATPase activity in two groups was lower than that in control group (P < 0.05). CONCLUSIONS: nNOS and iNOS play a role in cerebral mitochondrial structure and function damage in fetal rats with intrauterine distress. L-NNA has some limited treatment effect on mitochondrial damage when intrauterine distress induces acute fetal hypoxic-ischemic brain damage. AG plays an obvious treatment role in mitochondrial damage during reperfusion following ischemia. PMID- 15363355 TI - [Torsion of fallopian tube and ovary in children--report of 4 cases]. PMID- 15363356 TI - [Comparative study of ultrasonography and pathology of intrauterine septa]. PMID- 15363357 TI - [Sarcoma transformation of endometriosis of abdominal wall --a case report with literature review]. PMID- 15363358 TI - [Successful pregnancy outcome following early-gestational uterine surgery--a case report]. PMID- 15363359 TI - [Expression of leptin and tumor necrosis factor-alpha in placenta in pregnancy induced hypertension syndrome]. PMID- 15363360 TI - [Relation Between microsomal epoxide hydrolase polymorphism and susceptibility to ovarian epithelial cancer]. PMID- 15363361 TI - [Methotrexate-induced apoptosis of JAR cells of choriocarcinoma and expression of its related proteins]. PMID- 15363362 TI - [Effects of transferring good quality embryo and reducing numbers of embryo on the outcome of in vitro fertilization-embryo transfer]. PMID- 15363363 TI - [Surgical treatment of vaginal prolapses]. PMID- 15363364 TI - [Progress on the epidemiological study and treatment of female urinary incontinence]. PMID- 15363365 TI - [Progress in the study on correlation of expression of survivin gene and gynecologic tumors]. PMID- 15363366 TI - [Etiology of brachial plexus injury in neonate]. PMID- 15363367 TI - [Acardiac twin--a case report]. PMID- 15363368 TI - [Specific dermatoses of pregnancy]. PMID- 15363369 TI - [12th International annual meeting on Gynecologic Endoscopy]. PMID- 15363371 TI - A meta-analysis of outcome rating scales in foot and ankle surgery: is there a valid, reliable, and responsive system? AB - BACKGROUND: Rating scales that are valid, reliable, and responsive communicate the severity of a functional problem, facilitate the accurate study of treatment modalities, and provide a common language for those involved in research. The purpose of this study was to determine which outcome rating scales are currently used in the foot and ankle literature and to identify rating scales with proven reliability, validity, and responsiveness. METHOD: A meta-analysis of the foot and ankle literature from 1990 to 2001 was done. All referenced rating scales were reviewed to determine if any had proven to be reliable, valid, or responsive. RESULTS: Forty-nine different rating scales were identified. The most frequently referenced scales were the subscales of the American Orthopaedic Foot and Ankle Society (AOFAS). No rating scale was identified that demonstrated reliability, validity, and responsiveness in patients with a variety of foot and ankle conditions. CONCLUSIONS: The development of a reliable, valid, and responsive rating scale would have value not only in assessing patient outcomes but also in reporting the results of clinical studies in foot and ankle surgery. PMID- 15363372 TI - The Lapidus procedure: a prospective cohort outcome study. AB - BACKGROUND: Multiple reports are found in the literature about the surgical treatment of hallux valgus deformities. Few, however, are prospective outcome studies. The purpose of this study was to prospectively look at the functional outcome of patients with moderate and severe hallux valgus deformities (intermetatarsal angle of more than 14 degrees and a hallux valgus angle of more than 30 degrees) after the Lapidus procedure. METHODS: Indication for surgery and inclusion into the study was failure of nonoperative management for metatarsus primus varus and painful hallux valgus deformities. Data was collected using the AOFAS Hallux Metatarsophalangeal Interphalangeal Scale, Visual Analog Pain Scale, clinical examination, weightbearing radiographs, and a patient satisfaction questionnaire. Data was collected preoperatively, 6 weeks after surgery, 6 months after surgery, and then yearly. One hundred and five feet in 91 patients were followed for an average of 3.7 years (range, 18 months to 6.2 years). Sixty-two patients were female and 29 were male. The average age at surgery was 41 years (range, 20 years to 71 years). RESULTS: The AOFAS Hallux Metatarsophalangeal Interphalangeal Scale scores increased from 52 preoperatively to 87 points at most recent follow-up (p <.001). The average intermetatarsal angle preoperatively was 18 degrees and 8.2 degrees at most recent follow-up. Between the 1-year and 3.7-year follow-up visits, only.3-degree increase of the intermetatarsal angle was noted. The average hallux valgus angle preoperatively was 37 degrees and 16 degrees at most recent follow-up. Less than 1-degree increase in the hallux valgus angle was noted between the 1-year to 3.7-year follow-up visits. Complications included tarsometatarsal nonunion in seven patients. Five patients lost correction and all seven patients had a revision procedure done for pain. Removal of hardware was necessary in eight patients, minor wound problems occurred in two patients, superficial neuroma in two patients, and transfer metatarsalgia in four. CONCLUSIONS: The Lapidus procedure obviously is not the answer to all bunion deformities, but with proper technique and attention to detail it is an excellent alternative treatment for moderate to severe metatarsus primus varus and hallux valgus deformities. PMID- 15363373 TI - Clinical results with the Ludloff osteotomy for correction of adult hallux valgus. AB - BACKGROUND: Biomechanically, the Ludloff osteotomy fixed with lag screw compression has been shown to be more rigid than proximal crescentic and other proximal first metatarsal osteotomies for correction of symptomatic hallux valgus with a moderate to severe increase in the first intermetatarsal angle. The Ludloff osteotomy may, therefore, have a lower incidence of dorsal malunion and transfer metatarsalgia than other proximal first metatarsal osteotomies, such as the crescentic or chevron. METHODS: We reviewed the results of 82 consecutive cases of moderate to severe hallux valgus deformities corrected with the Ludloff oblique metaphyseal-diaphyseal osteotomy of the first metatarsal combined with a distal soft-tissue procedure and medial eminence resection. RESULTS: Follow-up was possible in 70 cases (85%) at an average of 30 months (range, 18 to 42 months). Preoperatively, the mean hallux valgus and first intermetatarsal angles were 31 degrees and 16 degrees, respectively. Postoperatively, these values improved to an average of 11 degrees and 7 degrees. In the sagittal plane, the first metatarsal was plantarflexed by an average of 1 mm, and there were no symptomatic transfer lesions of the second metatarsal. The mean AOFAS hindfoot score improved from 54 to 91 points. Complications included prominent hardware requiring removal (5), hallux varus (4), delayed union (3), superficial infection (3), and neuralgia (3). CONCLUSIONS: The use of the Ludloff oblique first metatarsal osteotomy resulted in excellent correction of the first intermetatarsal angle in patients with moderate to severe hallux valgus. With the plane of the osteotomy and rigidity of fixation, immediate ambulation was possible with minimal risk of dorsiflexion malunion of the first metatarsal. PMID- 15363374 TI - Hallux valgus and first ray mobility: a cadaveric study. AB - BACKGROUND: Several studies have demonstrated that patients with hallux valgus (HV) deformities have increased first ray sagittal mobility. However, the change in mobility that occurs after surgical correction of HV deformities has not been extensively evaluated. This study was done to determine if surgical realignment of the first ray in cadaver specimens with a proximal crescentic osteotomy and distal soft tissue reconstruction (DSTR) would reduce the first ray sagittal motion as measured with an external-type micrometer (the Klaue device). METHODS: Twelve fresh-frozen below-knee cadaver specimens with an HV deformity (HV angle > 15 degrees, 1-2 IM angle > 9 degrees) were used for the study. Standardized simulated weightbearing radiographs were obtained before and after the surgical correction of the deformity. The first ray sagittal motion was measured with an external micrometer (Klaue device) before correction of the HV deformity and after the procedure. All specimens had correction of the hallux valgus deformity with a DSTR and proximal crescentic osteotomy. Internal fixation was applied to secure the osteotomy site. RESULTS: The HV angle was corrected from a mean of 28.6 degrees to a mean of 11.0 degrees. The 1-2 IM angle was corrected from a mean of 12.9 degrees to a mean of 6.8 degrees. The average preoperative first ray sagittal motion was 11.0 mm (range, 8.5 mm to 13.5 mm). After the surgical repair, the mean sagittal first ray motion was significantly decreased (p <.0005) to a mean of 5.2 mm (range, 3.5 mm to 7.5 mm). CONCLUSION: After correction of HV deformities with a DSTR and a proximal crescentic osteotomy, first ray mobility in cadaver specimens was significantly reduced. The stabilization of first ray mobility that occurred immediately after surgical correction despite leaving the capsule of the first metatarsocuneiform (MC) joint undisturbed suggests that extrinsic anatomic features may play a role in first ray mobility. Additionally, stability of the first ray may be restored with a bunion procedure that does not sacrifice the first MC joint. PMID- 15363375 TI - Surgical versus accommodative treatment for Charcot arthropathy of the midfoot. AB - BACKGROUND: The treatment of Charcot foot arthropathy is one of the most controversial issues facing orthopaedic foot and ankle surgeons. Although current orthopaedic textbooks are in almost universal agreement that treatment should be nonoperative, accommodating the deformity with orthotic methods, most peer reviewed clinical studies recommend early surgical correction of the deformity. In a university health system orthopaedic foot and ankle clinic with a special interest in diabetic foot disorders, a moderate approach evolved for management of this difficult patient population. METHODS: Patients with Charcot arthropathy and plantigrade feet were treated with accommodative orthotic methods. Those with nonplantigrade feet were treated with surgical correction of the deformity, followed by long-term management with commercial therapeutic footwear. The desired outcome for both groups was long-term management with standard, commercially available, therapeutic depth-inlay shoes and custom-fabricated accommodative foot orthoses. During a 6-year period, 198 patients (201 feet) were treated for diabetes-associated Charcot foot arthropathy. The location of the deformity was in the midfoot in 147 feet, in the ankle in 50, and in the forefoot in four. RESULTS: At a minimum 1-year follow-up, 87 of the 147 feet with midfoot disease (59.2%) achieved the desired endpoint without surgical intervention. Sixty (40.8%) required surgery. Corrective osteotomy with or without arthrodesis was attempted in 42, while debridement or simple exostectomy was attempted in 18 feet. Three patients had initial amputation (one partial foot amputation, one Syme ankle disarticulation, and one transtibial amputation), and five had amputation (two Syme ankle disarticulations and three transtibial amputations) after attempted salvage failed. CONCLUSION: Using a simple treatment protocol with the desired endpoint being long-term management with commercially available, therapeutic footwear and custom foot orthoses, more than half of patients with Charcot arthropathy at the midfoot level can be successfully managed without surgery. PMID- 15363376 TI - Dorsal mobility and first ray stiffness in patients with diabetes mellitus. AB - BACKGROUND: Limited joint mobility in diabetic patients has been identified as a risk factor in the development of plantar ulcers. We examined dorsal mobility and passive first ray stiffness in patients with diabetes and investigated the relationship between first ray mobility and ankle joint dorsiflexion. METHODS: Forty individuals were studied: 20 with diabetes (mean estimated duration of 16 +/- 10 years) and 20 matched controls. Dorsal first ray mobility was measured using a mechanical device. Force-vs-dorsal mobility displacement values were collected at 10 N increments to a load limit of 55 N. Ankle joint dorsiflexion motion was measured with a goniometer. The "prayer sign," a clinical indicator of limited joint mobility, was evaluated in each patient. Subjects were separated into the two groups for data analysis. RESULTS: Patients in the diabetic group had more stiffness and less dorsal first ray mobility than the control group (p <.05). In particular, patients with a positive prayer sign had significant first ray stiffness (p <.05). Patients with diabetes also had less ankle dorsiflexion (p <.05). CONCLUSION: Patients with diabetes have more stiffness and less first ray mobility and less ankle dorsiflexion than those without diabetes. The presence of a positive prayer sign correlates with stiffness and loss of first ray mobility. Soft-tissue stiffness may contribute to the development of foot ulcers in diabetic patients with neuropathy. PMID- 15363377 TI - Safety of total contact casting in high-risk patients with neuropathic foot ulcers. AB - BACKGROUND: Total contact casting (TCC) is effective in offloading the plantar aspect of the foot in patients with diabetes and neuropathic ulcers. These patients are considered at high risk for skin-related complications during TCC because of sensory neuropathy. The purpose of this prospective study was to determine the frequency of complications during treatment of neuropathic ulcers with TCC. METHODS: Thirteen patients with 18 neuropathic ulcers were treated with TCC. The same orthopaedic surgeon applied a consecutive series of 82 total contact casts. The initial cast was changed in 3 to 4 days, while subsequent casts were changed weekly. RESULTS: Fourteen complications occurred during the 82 castings (17%). None of the complications required alteration in the treatment protocol. Thirteen of the 14 complications involved skin irritation and the other complication was from a cast that became too tight. Fifteen of the 18 neuropathic ulcers healed with TCC. CONCLUSIONS: TCC can be used safely in high-risk patients with neuropathic problems, but minor complications should be anticipated. Major complications that interfere with the treatment of the plantar ulcer can be minimized with careful technique, close follow-up, and thorough patient education. PMID- 15363378 TI - Use of a Torque-Range-of-Motion device for objective differentiation of diabetic from normal feet in adults. AB - BACKGROUND: The ability of the foot and ankle complex to act as an energy absorber is reflected in its viscoelastic properties. The Torque-Range-of-Motion (TROM) device was designed to provide an effective objective assessment of foot and ankle passive mechanical function. The hypothesis of this study was that mechanical parameters derived from passive TROM curves of otherwise normal feet of adults with diabetes would be significantly different from those of adults without diabetes. METHODS: The TROM device is a single-degree-of-freedom hinge transducer system that is manually rotated through plantarflexion and dorsiflexion. The device was rotated manually with the muscles relaxed during a 50-second data acquisition period. A strain gauge provided the torque signal and a precision single-turn potentiometer provided plantarflexion-dorsiflexion angle to a two-channel portable data acquisition system. With the TROM device connected to a computer, input for instantaneous torque and range of motion was acquired and displayed as angle (degrees) versus torque (Newton-meters) on an output screen. The period provided sufficient data to average 16 to 20 cycles of motion. The study included 41 feet in adults without diabetes and 42 age-matched feet in adults with diabetes but no known foot problems. RESULTS: For a probability level of.0001 there were significant differences in hysteresis area (normal: 91.1 +/- 46.9 Nm-deg and diabetic: 161.7 +/- 65.7 Nm-deg) and both dorsiflexion (normal: 0.4 +/- 0.1 Nm/deg and diabetic: 0.9 +/- 0.3 Nm/deg) and plantarflexion stiffness (normal: 0.3 +/- 0.1 Nm/deg and diabetic: 0.7 +/- 0.3 Nm/deg). CONCLUSIONS: The feet of adults with diabetes absorb more energy during cyclic motion (thus must dissipate more energy per cycle) and are stiffer in the terminal regions (where muscle-tendon-ligament properties prevail) than are the feet of adults without diabetes. These results suggest that this passive TROM method may be a sensitive, objective measurement of the viscoelastic properties of the foot and ankle, which may be an early indicator of diabetic patients who are at risk for the development of foot problems. PMID- 15363379 TI - Plantarflexion opening wedge medial cuneiform osteotomy for correction of fixed forefoot varus associated with flatfoot deformity. AB - BACKGROUND: Flatfoot presents as a wide spectrum of foot deformities that include varying degrees of hindfoot valgus, forefoot abduction, and forefoot varus. Medial displacement calcaneal osteotomy, lateral column lengthening, and subtalar fusion can correct heel valgus, but may not adequately correct the fixed forefoot varus component. The purpose of this study was to determine the effectiveness of plantarflexion opening wedge medial cuneiform (Cotton) osteotomy in the correction of forefoot varus. METHODS: Sixteen feet (15 patients) had plantarflexion opening wedge medial cuneiform osteotomies to correct forefoot varus associated with flatfoot deformities from several etiologies, including congenital flatfoot (six feet, average age 37 years), tarsal coalition (five feet, average age 15 years), overcorrected clubfoot deformity (two feet, ages 17 years and 18 years), skewfoot (one foot, age 15 years), chronic posterior tibial tendon insufficiency (one foot, 41 years), and rheumatoid arthritis (one foot, age 56 years). RESULTS: Standing radiographs showed an average improvement in the anterior-posterior talo-first metatarsal angle of 7 degrees (9 degrees preoperative, 2 degrees postoperative). The talonavicular coverage angle improved an average of 15 degrees (20 degrees preoperative, 5 degrees postoperative). The lateral talo-first metatarsal angle improved an average of 14 degrees (-13 degrees preoperative, 1 degree postoperative). Correcting for radiographic magnification, the distance from the mid-medial cuneiform to the floor on the lateral radiograph averaged 40 mm preoperatively and 47 mm postoperatively (average improvement 7 mm). All patients at followup described mild to no pain with ambulation. There were no nonunions or malunions. CONCLUSIONS: Opening wedge medial cuneiform osteotomy is an important adjunctive procedure to correct the forefoot varus component of a flatfoot deformity. Advantages of this technique in comparison to first tarsometatarsal arthrodesis include predictable union, preservation of first ray mobility, and the ability to easily vary the amount of correction. Because of the variety of hindfoot procedures done in these patients, the degree of hindfoot correction contributed by the cuneiform osteotomy alone could not be determined. We have had excellent results without major complications using this technique. PMID- 15363380 TI - The arch index: a measure of flat or fat feet? AB - BACKGROUND: Studies using footprint-based estimates of arch height have indicated that obesity results in a lowered medial longitudinal arch in children. However, the potentially confounding effect of body composition on indirect measures of arch height, such as the arch index, has not been investigated. METHODS: This study assessed the body composition of 12 male and 12 female adults (mean age: 39.9 +/- 8.1 years, height: 1.724 +/- 0.101 m; weight: 95.1 +/- 13.7 kg, and BMI: 31.9 +/- 3.0 kg/m(2)) using bioelectrical impedance analysis to produce a two component model of fat mass (FM) and fat-free mass (FFM). The dynamic arch index also was determined from electronic footprints captured during gait using a capacitive pressure distribution platform with a resolution of 4 sensors/cm(2). RESULTS: While significant correlations were noted between FFM and the area of both the hindfoot (r =.75, p <.05) and forefoot (r =.72, p <. 05), the midfoot area was correlated only with FM (r =.54, p <.05). Similarly, the arch index was significantly correlated with the FM percentage (r =.67, p <.05). CONCLUSIONS: The findings of this pilot study suggest that body composition influences arch index values in overweight and obese subjects. Consequently, body composition may be a confounding factor in interpreting footprint based estimates of arch height and, as such, these estimates would best be used with supplementary measures of body composition. PMID- 15363381 TI - Gender-specific differences of the foot during the first year of walking. AB - BACKGROUND: It is is well established that gender-specific differences in foot shape exist in adults. However, no information is available whether these differences are present already in children's feet and at what stage in the development these changes become evident. METHODS: Plantar pressure patterns and footprints of 42 healthy infants (20 male, 22 female) were evaluated over the course of one year and compared regarding gender-specific differences. The age at initial evaluation of the children was 16.1 months, and they were evaluated every 3 months. Anthropometric data, foot shape, and foot loading parameters for the whole foot, as well as for selected regions of interest, were evaluated. RESULTS: Significant differences in the foot shape and dynamic foot loading parameters were noted between boys and girls already at this young age. Boys demonstrated a broader midfoot, indicating a lower arch. Girls showed a more pronounced dynamic loading, especially in the heel and forefoot regions. CONCLUSIONS: The differences seen in this study should be taken into account by the shoe industry responsible for the design of children's shoes to support healthy development of the growing foot. Foot disorders are not only caused by hereditary factors but may be due to disturbances during foot growth and development. Since today's society is dependent on wearing shoes for foot protection, shoes should be designed not only according to the fashion demands of the customers but also to the biomechanical needs. Therefore, it is important to ensure that shoes fit the shape of the feet, which is different for boys and girls already when walking begins. PMID- 15363382 TI - The role of the interosseous talocalcaneal ligament in subtalar joint stability. AB - BACKGROUND: Injury of the interosseous talocalcaneal ligament (ITCL) has been recognized as a cause of subtalar instability, though lack of an accepted clinical test has limited the ability of clinicians to reliably make the diagnosis. Clinical effects of ITCL failure remain unclear because of insufficient understanding of the role of the ligament. METHODS: Load displacement characteristics of the subtalar joint were studied in six cadaver specimens using an axial distraction test and a transverse multi-direction drawer test. In all tests, cyclic loading (+/-60 N) was applied, and load-displacement responses were collected before and after sectioning of the ITCL. Two parameters were used to analyze the data: neutral-zone laxity as a measure of joint play, and flexibility as a measure of resistance to applied force. RESULTS: In the axial distraction test, sectioning increased both neutral-zone laxity and flexibility (p =.01 and.02, respectively). In the transverse test, sectioning caused increase of both neutral-zone laxity and flexibility (p <.001, for each). Neutral-zone laxity increased most greatly along an axis defined roughly by the posterior aspect of the fibula and the central region of the medial malleolus. Flexibility increased most in the medial direction (p <.05, for each). CONCLUSIONS: Results confirmed the role of the ITCL in maintaining apposition of the subtalar joint, as well as suggested its role in stabilizing the subtalar joint against drawer forces applied to the calcaneus from lateral to medial. The dominant direction of increased neutral-zone laxity described above suggests the optimal direction for detecting subtalar instability involved with ITCL injury. CLINICAL RELEVANCE: ITCL failure may result in subtalar instability and should be examined with a drawer force along the preferential axis roughly from the posterior aspect of the fibula to the central region of the medial malleolus. Further clinical evaluation is required to determine whether ITCL failure is reliably detectable. PMID- 15363383 TI - Special report: highlights of the First Annual Scientific Meeting of the Canadian Orthopaedic Foot and Ankle Society, Winnipeg, Manitoba, October 4, 2003. PMID- 15363384 TI - Recurrent dislocation of the posterior tibial tendon secondary to detachment of a retinacular-periosteal sleeve: a case report. PMID- 15363385 TI - Re: Hallux rigidus: demographics, etiology, and radiographic assessment. Coughlin, MJ; Shurnas, PS, Foot Ankle Int. 24(10): 731-734, 2003. PMID- 15363386 TI - Out-foxing fate; molecular switches create neuronal diversity in the retina. AB - Progenitor cells in the mammalian retina generate at least 55 distinct kinds of neurons. Two reports in this issue of Neuron reveal two transcription factors (Foxn4 and Bhlhb4) that contribute to the development of this remarkable cellular diversity. PMID- 15363387 TI - Steering growth cones with a CaMKII/calcineurin switch. AB - Calcium can regulate and induce both attractive and repulsive turnings by growth cones. In this issue of Neuron, Wen et al. report that differential activations of CaMKII and calcineurin (CaN) act as the read out for distinct patterns of intracellular calcium signals and a switch between attraction and repulsion. PMID- 15363388 TI - Endocannabinoids: losing inhibition to increase learning capacity? AB - Recent work has implicated endocannabinoids in various forms of synaptic plasticity. In this issue of Neuron, Chevaleyre and Castillo describe a new mechanism whereby a CB1 receptor-mediated LTD of inhibitory synaptic transmission facilitates the subsequent induction of LTP in a narrow band of synapses surrounding a region of potentiated synapses. PMID- 15363389 TI - Cortical motion deafness. AB - The extent to which the auditory system, like the visual system, processes spatial stimulus characteristics such as location and motion in separate specialized neuronal modules or in one homogeneously distributed network is unresolved. Here we present a patient with a selective deficit for the perception and discrimination of auditory motion following resection of the right anterior temporal lobe and the right posterior superior temporal gyrus (STG). Analysis of stimulus identity and location within the auditory scene remained intact. In addition, intracranial auditory evoked potentials, recorded preoperatively, revealed motion-specific responses selectively over the resected right posterior STG, and electrical cortical stimulation of this region was experienced by the patient as incoming moving sounds. Collectively, these data present a patient with cortical motion deafness, providing evidence that cortical processing of auditory motion is performed in a specialized module within the posterior STG. PMID- 15363390 TI - The transcription factor Bhlhb4 is required for rod bipolar cell maturation. AB - Retinal bipolar cells are essential to the transmission of light information. Although bipolar cell dysfunction can result in blindness, little is known about the factors required for bipolar cell development and functional maturation. The basic helix-loop-helix (bHLH) transcription factor Bhlhb4 was found to be expressed in rod bipolar cells (RB). Electroretinograms (ERGs) in the adult Bhlhb4 knockout (Bhlhb4(-/-)) showed that the loss of Bhlhb4 resulted in disrupted rod signaling and profound retinal dysfunction resembling human congenital stationary night blindness (CSNB), characterized by the loss of the scotopic ERG b-wave. A depletion of inner nuclear layer (INL) cells in the adult Bhlhb4 knockout has been ascribed to the abolishment of the RB cell population during postnatal development. Other retinal cell populations including photoreceptors were unaltered. The timing of RB cell depletion in the Bhlhb4(-/-) mouse suggests that Bhlhb4 is essential for RB cell maturation. PMID- 15363392 TI - Development of morphological diversity of dendrites in Drosophila by the BTB-zinc finger protein abrupt. AB - Morphological diversity of dendrites contributes to specialized functions of individual neurons. In the present study, we examined the molecular basis that generates distinct morphological classes of Drosophila dendritic arborization (da) neurons. da neurons are classified into classes I to IV in order of increasing territory size and/or branching complexity. We found that Abrupt (Ab), a BTB-zinc finger protein, is expressed selectively in class I cells. Misexpression of ab in neurons of other classes directed them to take the appearance of cells with smaller and/or less elaborated arbors. Loss of ab functions in class I neurons resulted in malformation of their typical comb-like arbor patterns and generation of supernumerary branch terminals. Together with the results of monitoring dendritic dynamics of ab-misexpressing cells or ab mutant ones, all of the data suggested that Ab endows characteristics of dendritic morphogenesis of the class I neurons. PMID- 15363391 TI - Foxn4 controls the genesis of amacrine and horizontal cells by retinal progenitors. AB - During vertebrate retinogenesis, seven classes of cells are specified from multipotent progenitors. To date, the mechanisms underlying multipotent cell fate determination by retinal progenitors remain poorly understood. Here, we show that the Foxn4 winged helix/forkhead transcription factor is expressed in a subset of mitotic progenitors during mouse retinogenesis. Targeted disruption of Foxn4 largely eliminates amacrine neurons and completely abolishes horizontal cells, while overexpression of Foxn4 strongly promotes an amacrine cell fate. These results indicate that Foxn4 is both necessary and sufficient for commitment to the amacrine cell fate and is nonredundantly required for the genesis of horizontal cells. Furthermore, we provide evidence that Foxn4 controls the formation of amacrine and horizontal cells by activating the expression of the retinogenic factors Math3, NeuroD1, and Prox1. Our data suggest a model in which Foxn4 cooperates with other key retinogenic factors to mediate the multipotent differentiation of retinal progenitors. PMID- 15363393 TI - BTB/POZ-zinc finger protein abrupt suppresses dendritic branching in a neuronal subtype-specific and dosage-dependent manner. AB - How dendrites of different neuronal subtypes exhibit distinct branching patterns during development remains largely unknown. Here we report the mapping and identification of loss-of-function mutations in the abrupt (ab) gene that increased the number of dendritic branches of multiple dendritic (MD) sensory neurons in Drosophila embryos. Ab encodes an evolutionarily conserved transcription factor that contains a BTB/POZ domain and C2H2 zinc finger motifs. We show that ab has a cell-autonomous function in postmitotic neurons to limit dendritic branching. Ab and the homeodomain protein Cut are expressed in distinct but complementary subsets of MD neurons, and Ab functions in a transcriptional program that does not require Cut. Deleting one copy of ab or overexpressing ab had opposite effects on the formation of higher-order dendritic branches, suggesting that the Ab level in a specific neuron directly regulates dendritic complexity. These results demonstrate that dendritic branching can be suppressed by neuronal subtype-specific transcription factors in a cell-autonomous and dosage-dependent manner. PMID- 15363394 TI - A CaMKII/calcineurin switch controls the direction of Ca(2+)-dependent growth cone guidance. AB - Axon pathfinding depends on attractive and repulsive turning of growth cones to extracellular cues. Localized cytosolic Ca2+ signals are known to mediate the bidirectional responses, but downstream mechanisms remain elusive. Here, we report that calcium-calmodulin-dependent protein kinase II (CaMKII) and calcineurin (CaN) phosphatase provide a switch-like mechanism to control the direction of Ca(2+)-dependent growth cone turning. A relatively large local Ca2+ elevation preferentially activates CaMKII to induce attraction, while a modest local Ca2+ signal predominantly acts through CaN and phosphatase-1 (PP1) to produce repulsion. The resting level of intracellular Ca2+ concentrations also affects CaMKII/CaN operation: a normal baseline allows distinct turning responses to different local Ca2+ signals, while a low baseline favors CaN-PP1 activation for repulsion. Moreover, the cAMP pathway negatively regulates CaN-PP1 signaling to inhibit repulsion. Finally, CaMKII/CaN-PP1 also mediates netrin-1 guidance. Together, these findings establish a complex Ca2+ mechanism that targets the balance of CaMKII/CaN-PP1 activation to control distinct growth cone responses. PMID- 15363395 TI - Protein kinase CK2 is coassembled with small conductance Ca(2+)-activated K+ channels and regulates channel gating. AB - Small conductance Ca(2+)-activated K+ channels (SK channels) couple the membrane potential to fluctuations in intracellular Ca2+ concentration in many types of cells. SK channels are gated by Ca2+ ions via calmodulin that is constitutively bound to the intracellular C terminus of the channels and serves as the Ca2+ sensor. Here we show that, in addition, the cytoplasmic N and C termini of the channel protein form a polyprotein complex with the catalytic and regulatory subunits of protein kinase CK2 and protein phosphatase 2A. Within this complex, CK2 phosphorylates calmodulin at threonine 80, reducing by 5-fold the apparent Ca2+ sensitivity and accelerating channel deactivation. The results show that native SK channels are polyprotein complexes and demonstrate that the balance between kinase and phosphatase activities within the protein complex shapes the hyperpolarizing response mediated by SK channels. PMID- 15363396 TI - The super-cooling agent icilin reveals a mechanism of coincidence detection by a temperature-sensitive TRP channel. AB - TRPM8, a member of the transient receptor potential family of ion channels, depolarizes somatosensory neurons in response to cold. TRPM8 is also activated by the cooling agents menthol and icilin. When exposed to menthol or cold, TRPM8 behaves like many ligand-gated channels, exhibiting rapid activation followed by moderate Ca(2+)-dependent adaptation. In contrast, icilin activates TRPM8 with extremely variable latency followed by extensive desensitization, provided that calcium is present. Here, we show that, to achieve full efficacy, icilin requires simultaneous elevation of cytosolic Ca2+, either via permeation through TRPM8 channels or by release from intracellular stores. Thus, two stimuli must be paired to elicit full channel activation, illustrating the potential for coincidence detection by TRP channels. Determinants of icilin sensitivity map to a region of TRPM8 that corresponds to the capsaicin binding site on the noxious heat receptor TRPV1, suggesting a conserved molecular logic for gating of these thermosensitive channels by chemical agonists. PMID- 15363397 TI - Endocannabinoid-mediated metaplasticity in the hippocampus. AB - Repetitive activation of glutamatergic fibers that normally induces long-term potentiation (LTP) at excitatory synapses in the hippocampus also triggers long term depression at inhibitory synapses (I-LTD) via retrograde endocannabinoid signaling. Little is known, however, about the physiological significance of I LTD. Here, we show that synaptic-driven release of endocannabinoids is a highly localized and efficient process that strongly depresses cannabinoid-sensitive inhibitory inputs within the dendritic compartment of CA1 pyramidal cells. By removing synaptic inhibition in a restricted area of the dendritic tree, endocannabinoids selectively "primed" nearby excitatory synapses, thereby facilitating subsequent induction of LTP. This induction of local metaplasticity is a novel mechanism by which endocannabinoids can contribute to the storage of information in the brain. PMID- 15363398 TI - Oscillatory entrainment of striatal neurons in freely moving rats. AB - Oscillations and synchrony in basal ganglia circuits may play a key role in the organization of voluntary actions and habits. We recorded single units and local field potentials from multiple striatal and cortical locations simultaneously, over a range of behavioral states. We observed opposite gradients of oscillatory entrainment, with dorsal/lateral striatal neurons entrained to high-voltage spindle oscillations ("spike wave discharges") and ventral/medial striatal neurons entrained to the hippocampal theta rhythm. While the majority of units were likely medium-spiny projection neurons, a second neuronal population showed characteristic features of fast-spiking GABAergic interneurons, including tonic activity, brief waveforms, and high-frequency bursts. These fired at an earlier spindle phase than the main neuronal population, and their density within striatum corresponded closely to the intensity of spindle oscillations. The orchestration of oscillatory activity by networks of striatal interneurons may be an important mechanism in the pathophysiology of neurological disorders such as Parkinson's disease. PMID- 15363399 TI - Extinction learning in humans: role of the amygdala and vmPFC. AB - Understanding how fears are acquired is an important step in translating basic research to the treatment of fear-related disorders. However, understanding how learned fears are diminished may be even more valuable. We explored the neural mechanisms of fear extinction in humans. Studies of extinction in nonhuman animals have focused on two interconnected brain regions: the amygdala and the ventral medial prefrontal cortex (vmPFC). Consistent with animal models suggesting that the amygdala is important for both the acquisition and extinction of conditioned fear, amygdala activation was correlated across subjects with the conditioned response in both acquisition and early extinction. Activation in the vmPFC (subgenual anterior cingulate) was primarily linked to the expression of fear learning during a delayed test of extinction, as might have been expected from studies demonstrating this region is critical for the retention of extinction. These results provide evidence that the mechanisms of extinction learning may be preserved across species. PMID- 15363400 TI - Competing global representations fail to initiate binocular rivalry. AB - A longstanding debate in binocular rivalry literature is whether the perceptual competition in rivalry occurs at an early or late stage of visual processing. Central to this debate is the determination of the source of the competition. Overwhelming evidence exists that local interocular differences can lead to binocular rivalry, but it is not yet clear whether interocular conflicts at the global level are sufficient to generate binocular rivalry. The current study adopted a novel stimulus that enabled the introduction of dramatic global differences between the two eyes with compatible local elements. Results show that global differences between the two eyes' images do not result in rivalry if local elements are compatible. The implication of these findings is that the registration of competing interocular information, necessary to generate binocular rivalry, is performed at an early stage of visual processing prior to global analysis of the image. PMID- 15363401 TI - Clathrin: now you see me, now you don't! AB - Endocytic clathrin-coated vesicles are short-lived transport intermediates that ferry cargo macromolecules rapidly into the cell interior. Recent work from the Kirchhausen laboratory indicates that the lifetime of a coated vesicle is extremely short, and assembly of nascent coats aborts abruptly unless reinforced by additional regulatory inputs, most likely cargo capture. PMID- 15363402 TI - Surfing the actomyosin wave: polarization of the C. elegans zygote. AB - Many cell types rely on asymmetrically localized PAR proteins to become polarized. New evidence reveals that cortical flows powered by actomyosin contractions can mobilize PAR complexes to create distinct cortical domains. PMID- 15363403 TI - Lipid phase perturbations and the unfolded protein response. AB - Recent studies of the consequences of ganglioside accumulation in lysosomal storage disease and free cholesterol accumulation in cell membranes in atherosclerosis suggest an unexpected link between perturbation of the endoplasmic reticulum membrane's lipid phase, induction of the unfolded protein response, and cell death. PMID- 15363404 TI - ReFUSing to grow up. AB - At germination, plants must coordinate the exit from dormancy and the start of vegetative growth. In this issue of Developmental Cell, Gazzarrini et al. show that FUSCA3 plays a critical role in regulating hormone levels to synchronize the transition from embryonic to vegetative growth in Arabidopsis. PMID- 15363405 TI - Gene-regulatory interactions in neural crest evolution and development. AB - In this review, we outline the gene-regulatory interactions driving neural crest development and compare these to a hypothetical network operating in the embryonic ectoderm of the cephalochordate amphioxus. While the early stages of ectodermal patterning appear conserved between amphioxus and vertebrates, later activation of neural crest-specific factors at the neural plate border appears to be a vertebrate novelty. This difference may reflect co-option of genetic pathways which conferred novel properties upon the evolving vertebrate neural plate border, potentiating the evolution of definitive neural crest. PMID- 15363406 TI - Segmentation. AB - The three major taxa with metameric segmentation (annelids, arthropods, and chordates) appear to use three very different molecular strategies to generate segments. However, unexpected similarities are starting to emerge from characterization of pair-rule patterning and segmental border formation. Moreover, the existence of an ancestral segmentation clock based on Notch signaling has become likely. An old concept of comparative anatomy, the enterocoele theory, is compatible with a single origin of segmentation mechanisms and could therefore provide a conceptual framework for assessing these molecular similarities. PMID- 15363407 TI - The unicellular ancestry of animal development. AB - The transition to multicellularity that launched the evolution of animals from protozoa marks one of the most pivotal, and poorly understood, events in life's history. Advances in phylogenetics and comparative genomics, and particularly the study of choanoflagellates, are yielding new insights into the biology of the unicellular progenitors of animals. Signaling and adhesion gene families critical for animal development (including receptor tyrosine kinases and cadherins) evolved in protozoa before the origin of animals. Innovations in transcriptional regulation and expansions of certain gene families may have allowed the integration of cell behavior during the earliest experiments with multicellularity. The protozoan perspective on animal origins promises to provide a valuable window into the distant past and into the cellular bases of animal development. PMID- 15363408 TI - Development and ecology in the time of systems biology. PMID- 15363409 TI - Foxh1 is essential for development of the anterior heart field. AB - The anterior heart field (AHF) mediates formation of the outflow tract (OFT) and right ventricle (RV) during looping morphogenesis of the heart. Foxh1 is a forkhead DNA binding transcription factor in the TGFbeta-Smad pathway. Here we demonstrate that Foxh1-/- mutant mouse embryos form a primitive heart tube, but fail to form OFT and RV and display loss of outer curvature markers of the future working myocardium, similar to the phenotype of Mef2c-/- mutant hearts. Further, we show that Mef2c is a direct target of Foxh1, which physically and functionally interacts with Nkx2-5 to mediate strong Smad-dependent activation of a TGFbeta response element in the Mef2c gene. This element directs transgene expression to the presumptive AHF, as well as the RV and OFT, a pattern that closely parallels endogenous Mef2c expression in the heart. Thus, Foxh1 and Nkx2-5 functionally interact and are essential for development of the AHF and its derivatives, the RV and OFT, in response to TGFbeta-like signals. PMID- 15363410 TI - Tsukushi functions as an organizer inducer by inhibition of BMP activity in cooperation with chordin. AB - During chick gastrulation, inhibition of BMP signaling is required for primitive streak formation and induction of Hensen's node. We have identified a unique secreted protein, Tsukushi (TSK), which belongs to the Small Leucine-Rich Proteoglycan (SLRP) family and is expressed in the primitive streak and Hensen's node. Grafts of cells expressing TSK in combination with the middle primitive streak induce an ectopic Hensen's node, while electroporation of TSK siRNA inhibits induction of the node. In Xenopus embryos, TSK can block BMP function and induce a secondary dorsal axis, while it can dorsalize ventral mesoderm and induce neural tissue in embryonic explants. Biochemical analysis shows that TSK binds directly to both BMP and chordin and forms a ternary complex with them. These observations indicate that TSK is an essential dorsalizing factor involved in the induction of Hensen's node. PMID- 15363411 TI - A role of VAMP8/endobrevin in regulated exocytosis of pancreatic acinar cells. AB - Despite our general understanding that members of the SNARE superfamily participate in diverse intracellular docking/fusion events, the physiological role of the majority of SNAREs in the intact organism remains elusive. In this study, through targeted gene knockout in mice, we establish that VAMP8/endobrevin is a major player in regulated exocytosis of the exocrine pancreas. VAMP8 is enriched on the membrane of zymogen granules and exists in a complex with syntaxin 4 and SNAP-23. VAMP8-/- mice developed normally but showed severe defects in the pancreas. VAMP8 null acinar cells contained three times more zymogen granules than control acinar cells. Furthermore, secretagogue-stimulated secretion was abolished in pancreatic fragments derived from VAMP8-/- mice. In addition, VAMP8-/- mice were partially resistant to supramaximal caerulein induced pancreatitis. These results suggest a major physiological role of VAMP8 in regulated exocytosis of pancreatic acinar cells by serving as a v-SNARE of zymogen granules. PMID- 15363412 TI - The transcription factor FUSCA3 controls developmental timing in Arabidopsis through the hormones gibberellin and abscisic acid. AB - Although plants continually produce different organs throughout their life cycle, little is known about the factors that regulate the timing of a given developmental program. Here we report that the restricted expression of FUS3 to the epidermis is sufficient to control foliar organ identity in Arabidopsis by regulating the synthesis of two hormones, abscisic acid and gibberellin. These hormones in turn regulate the rates of cell cycling during organ formation to determine whether an embryonic or adult leaf will emerge. We also show that FUS3 expression is influenced by the patterning hormone, auxin, and therefore acts as a nexus of hormone action during embryogenesis. The identification of lipophillic hormones downstream of a heterochronic regulator in Arabidopsis has parallels to mechanisms of developmental timing in animals and suggests a common logic for temporal control of developmental programs between these two kingdoms. PMID- 15363413 TI - Invasive cell behavior during Drosophila imaginal disc eversion is mediated by the JNK signaling cascade. AB - Drosophila imaginal discs are monolayered epithelial invaginations that grow during larval stages and evert at metamorphosis to assemble the adult exoskeleton. They consist of columnar cells, forming the imaginal epithelium, as well as squamous cells, which constitute the peripodial epithelium and stalk (PS). Here, we uncover a new morphogenetic/cellular mechanism for disc eversion. We show that imaginal discs evert by apposing their peripodial side to the larval epidermis and through the invasion of the larval epidermis by PS cells, which undergo a pseudo-epithelial-mesenchymal transition (PEMT). As a consequence, the PS/larval bilayer is perforated and the imaginal epithelia protrude, a process reminiscent of other developmental events, such as epithelial perforation in chordates. When eversion is completed, PS cells localize to the leading front, heading disc expansion. We found that the JNK pathway is necessary for PS/larval cells apposition, the PEMT, and the motile activity of leading front cells. PMID- 15363414 TI - Directional cell migration establishes the axes of planar polarity in the posterior lateral-line organ of the zebrafish. AB - The proper orientation of mechanosensory hair cells along the lateral-line organ of a fish or amphibian is essential for the animal's ability to sense directional water movements. Within the sensory epithelium, hair cells are polarized in a stereotyped manner, but the mechanisms that control their alignment relative to the body axes are unknown. We have found, however, that neuromasts can be oriented either parallel or perpendicular to the anteroposterior body axis. By characterizing the strauss mutant zebrafish line and by tracking labeled cells, we have demonstrated that neuromasts of these two orientations originate from, respectively, the first and second primordia. Furthermore, altering the migratory pathway of a primordium reorients a neuromast's axis of planar polarity. We propose that the global orientation of hair cells relative to the body axes is established through an interaction between directional movement by primordial cells and the timing of neuromast maturation. PMID- 15363415 TI - Cortical flows powered by asymmetrical contraction transport PAR proteins to establish and maintain anterior-posterior polarity in the early C. elegans embryo. AB - The C. elegans PAR proteins PAR-3, PAR-6, and PKC-3 are asymmetrically localized and have essential roles in cell polarity. We show that the one-cell C. elegans embryo contains a dynamic and contractile actomyosin network that appears to be destabilized near the point of sperm entry. This asymmetry initiates a flow of cortical nonmuscle myosin (NMY-2) and F-actin toward the opposite, future anterior, pole. PAR-3, PAR-6, and PKC-3, as well as non-PAR proteins that associate with the cytoskeleton, appear to be transported to the anterior by this cortical flow. In turn, PAR-3, PAR-6, and PKC-3 modulate cortical actomyosin dynamics and promote cortical flow. PAR-2, which localizes to the posterior cortex, inhibits NMY-2 from accumulating at the posterior cortex during flow, thus maintaining asymmetry by preventing inappropriate, posterior-directed flows. Similar actomyosin flows accompany the establishment of PAR asymmetries that form after the one-cell stage, suggesting that actomyosin-mediated cortical flows have a general role in PAR asymmetry. PMID- 15363416 TI - Mesenchymal-epithelial transition during somitic segmentation is regulated by differential roles of Cdc42 and Rac1. AB - Mesenchymal-epithelial transitions (MET) are crucial for vertebrate organogenesis. The roles of Rho family GTPases in such processes during actual development remain largely unknown. By electroporating genes into chick presomitic mesenchymal cells, we demonstrate that Cdc42 and Rac1 play important and different roles in the MET that generates the vertebrate somites. Presomitic mesenchymal cells, which normally contribute to both the epithelial and mesenchymal populations of the somite, were hyperepithelialized when Cdc42 signaling was blocked. Conversely, cells taking up genes that elevate Cdc42 levels remained mesenchymal. Thus, Cdc42 activity levels appear critical for the binary decision that defines the epithelial and mesenchymal somitic compartments. Proper levels of Rac1 are necessary for somitic epithelialization, since cells with activated or inhibited Rac1 failed to undergo correct epithelialization. Furthermore, Rac1 appears to be required for Paraxis to act as an epithelialization-promoting transcription factor during somitogenesis. PMID- 15363417 TI - Hmx2 and Hmx3 homeobox genes direct development of the murine inner ear and hypothalamus and can be functionally replaced by Drosophila Hmx. AB - The Hmx homeobox gene family appears to play a conserved role in CNS development in all animal species examined, and in higher vertebrates has an additional role in sensory organ development. Here, we show that murine Hmx2 and Hmx3 have both overlapping and distinct functions in the development of the inner ear's vestibular system, whereas their functions in the hypothalamic/pituitary axis of the CNS appear to be interchangeable. As in analogous knockin studies of Otx and En function, Drosophila Hmx can rescue conserved functions in the murine CNS. However, in contrast to Otx and En, Drosophila Hmx also rescues significant vertebrate-specific functions outside the CNS. Our work suggests that the evolution of the vertebrate inner ear may have involved (1) the redeployment of ancient Hmx activities to regulate the cell proliferation of structural components and (2) the acquisition of additional, vertebrate-specific Hmx activities to regulate the sensory epithelia. PMID- 15363418 TI - Accountability of the accused: facing public perceptions about financial conflicts of interest in spine surgery. PMID- 15363419 TI - The effect of spinal steroid injections for degenerative disc disease. AB - BACKGROUND: No conclusive evidence exists to determine that spinal steroid injections give lasting improvement in patients with predominantly axial low back pain resulting from lumbar degenerative disc disease (DDD). PURPOSE: The objectives of the study were to determine the effect of epidural steroid injections (ESIs) and intradiscal steroid injections (ISIs) in patients who exhibit DDD symptoms for more than 1 year and to determine whether patients with inflammatory end-plate changes are a unique subgroup of DDD patients in terms of treatment response. STUDY DESIGN: Pain and function in patients with DDD were prospectively assessed by an outcomes questionnaire before and after various spinal injections. Further correlation was made with end-plate inflammatory (Modic Type 1) changes identified on magnetic resonance imaging (MRI). PATIENT SAMPLE: ESI was performed in 232 patients who were referred for treatment of DDD, and discography with or without intradiscal steroid was performed in 171 patients who were possible spinal arthrodesis candidates. OUTCOME MEASURES: Pain and function were determined by a self-administered outcomes questionnaire that consisted of a visual analog pain scale, pain drawing, Oswestry Disability Index, use of pain medication and opinion of treatment success. METHODS: ESI was performed in 93 patients with DDD and inflammatory end-plate changes and in 139 patients without inflammatory end-plate changes. Patients with inflammatory end plate changes (n=78) or without inflammatory end-plate changes (n=93), all of whom were considered fusion candidates, underwent discography with or without intradiscal steroid in a randomized fashion. Pain and function were prospectively determined by a self-administered outcomes survey (VAS pain, Oswestry Disability index [ODI], pain diagram [PD] and opinion of success) before and after the patients' injection for a 2-year follow-up period. MRI and discography results were correlated with patient outcomes scores. RESULTS: ESI was effective in improving pain and function, as assessed by outcomes scores at short-term follow up. However, at 2 years, less than one-third had not had additional invasive treatment. Patients with inflammatory end-plate changes had greater improvement in ODI and PD scores in the first 6 months than did those patients without the end-plate changes. Intradiscal steroid injections into discs with concordant pain at the time of discography led to significant improvement in patients with inflammatory end-plate changes in all outcomes scales, but only minimal temporary improvement in patients without the end-plate changes. Disc pressure manometry at the time of discography found that discs with adjacent inflammatory end-plate changes reproduced symptoms at pressures significantly lower than those in other types of discs. CONCLUSIONS: Spinal steroid injections, both ESI and ISI, are beneficial for a small number of patients with advanced DDD and chronic low back pain. For those patients in whom a beneficial effect is found, spinal steroid injection is a low-risk and rapid treatment option. Spinal steroid injections are more effective in patients with MRI findings of discogenic inflammation, specifically adjacent inflammatory end-plate changes. PMID- 15363420 TI - Analysis of patient variables affecting neurologic outcome after traumatic cervical facet dislocation. AB - BACKGROUND CONTEXT: Traumatic cervical facet dislocation accounts for a disproportionate rate of neurologic disability. The relative importance of patient and management variables, including the timing of spinal reduction, in ultimate neurologic outcome has not been well defined. PURPOSE: To analyze data from a cohort of patients sustaining traumatic cervical facet dislocation to determine the relative importance of several patient and management variables in neurologic recovery after injury. STUDY DESIGN/SETTING: A retrospective study was conducted at a major referral center for spinal-cord-injured patients. PATIENT SAMPLE: Forty-five patients sustaining traumatic cervical facet dislocation. OUTCOME MEASURES: Using improvement in American Spinal Injury Association (ASIA) motor score as the primary outcome measure, patient data were used to construct a statistical model allowing the analysis of several clinically relevant variables. METHODS: The records of patients sustaining a traumatic cervical facet dislocation over a 5-year period were reviewed. Clinical data were collected for all patients with adequate follow-up. The data were used to construct a statistical model designed to analyze the contribution of the variables age, gender, time to reduction of the spine and initial motor score to neurologic improvement (the outcome measure). In addition, the effect of variable interaction was studied. RESULTS: Most patients demonstrated neurologic improvement over the course of follow-up after cervical facet dislocation. For this data set, the variables age and initial motor score were significantly associated with neurologic improvement. However, time to reduction of the spine did not demonstrate a significant independent relationship to neurologic outcome. No significant interaction was found between patient age or gender and the time to reduction with regard to predicting neurologic recovery. CONCLUSION: The present study uses a statistical model to determine the relative importance of clinically relevant variables for a population of patients after traumatic cervical facet dislocation. This model confirms the clinical impression that younger patients with lesser degrees of neurologic injury tend to achieve the best neurologic recovery after a traumatic facet dislocation. Although a strong benefit from earlier spinal column reduction did not emerge from the present data set, additional study is needed to define those patients who would benefit from immediate reduction of the spinal column. PMID- 15363421 TI - Pedicle screw fixation strength: pullout versus insertional torque. AB - BACKGROUND CONTEXT: Researchers studying early pedicle screw designs have suggested that pullout strength and insertional torque are correlated. For surgeons using pedicle screws, insertional torque is widely believed to be a good predictor of pullout strength and initial stability of the screw and construct. How appropriate is this assumption when applied to new screw and thread designs? PURPOSE: This study investigated the correlation between insertional torque and pullout strength of three different pedicle screw designs, with different insertional torque characteristics. We hypothesized that a significant increase in insertional torque would indicate a commensurate increase in pullout strength. STUDY DESIGN: Biomechanical analysis of instrumented vertebral specimens. METHODS: Calf lumbar vertebra were prepared and instrumented with one of three pedicle screws. Pilot hole preparation was standardized and coaxial orientation was confirmed by direct inspection. Screws did not penetrate the pedicle cortex or abut or penetrate the anterior vertebral cortex. Any specimen with pedicle wall breach was discarded. The pedicles were instrumented with one of three screws: 1) 7.5 x 40 mm conical, asymmetric progressive thread (Xia; Stryker Spine, Allendale, NJ), 2) 7.5 x 40 mm conical with traditional V-shaped thread (Osteonics, Stryker Spine, Allendale, NJ)) or 3) 6.5 x 40 mm cylindrical with V thread (Osteonics, cylindrical). Paired testing allowed individual screws to be directly compared with a contralateral "control." Insertional torque and peak torque values were recorded for each rotation up to full insertion. Pullout testing was conducted at a rate of 1 mm/minute. Load-displacement data were recorded at 20 Hz. Stiffness was considered the slope of the most linear part of the curve before the yield point. RESULTS: Peak loads for 7.5 conical Xia screws measured 1,783+/-589.1 N compared with 1,943.0+/-625.8 N for 7.5 conical Osteonics screws and 1,641.0+/-356.7 N for 6.5 cylindrical Osteonics screws. The peak insertional torque values were 6.7+/-1.9 Nm (158% greater than control), 4.5+/-1.1 Nm (73% greater than control) and 2.6+/-0.7 Nm, respectively. Insertional torques for Xia screws were significantly greater than conical (p=.001) and cylindrical Osteonics screws (p<.0001), and insertional torques for Osteonics conical screws were significantly greater than those of cylindrical screws (p<.0001). Although pullout loads for the conical Osteonics screws were consistently higher than either the Xia or Osteonics cylindrical screws, the differences were not significant (p>.05). There was no significant correlation between pullout strength and insertional torque (p>.05). CONCLUSIONS: This unexpected result is best explained by the progressively narrowing flutes of the Xia screw, which compact the trabeculae into a smaller volume as the screw nears full insertion. The trapezoidal threads also increase contact with the cortical surface area and compress trabeculae toward the cortex, thus creating greater friction and higher torque values. This increase in torque did not translate into a commensurate increase in pullout strength, where trabeculae fail in shear. PMID- 15363422 TI - Open versus endoscopic lumbar pedicle screw fixation and posterolateral fusion in a sheep model: a feasibility study. AB - BACKGROUND CONTEXT: Conventional open procedures for lumbar pedicle screw instrumentation are often associated with significant morbidity resulting in longer postoperative recovery and hence have led to the advent of less invasive techniques of spinal instrumentation and fusion. PURPOSE: This study compares lumbar pedicle screw fixation and posterolateral fusion using endoscopic and open techniques in a sheep model. STUDY DESIGN: Endoscopic and open techniques of lumbar pedicle screw instrumentation and posterolateral fusion are compared in a sheep model (12 sheep in each group). The safety and fusion outcome are assessed by postoperative computed tomography (CT) scans performed at 6 months. METHODS: Twelve sheep underwent bilateral L4-L5 pedicle screw fixation and posterolateral fusion by means of endoscopic instrumentation and another 12 sheep by means of conventional open technique. For posterolateral fusions, an autologous iliac crest bone or bone graft substitute (Healos/MP52; Orquest, Mountain View, CA) was used on either the animal's left or right side. The animals were euthanized 6 months after surgery and the lumbar spines were harvested. Pedicle dimensions and screw insertion angles were measured and the two techniques were compared using CT and image analysis software. Pedicle screw misplacement was detected by means of direct inspection and CT scan, and fusions were evaluated by CT scans. RESULTS: The endoscopic group's screw insertion angle was more convergent, corresponded more closely to the human pedicle angle and was less variable than in the open technique. Three of the endoscopic group's 48 screws were misplaced medially, whereas in the open technique, 5 screws were misplaced laterally. Posterolateral bone fusion masses were palpable 6 months after surgery for both the autologous bone grafts and graft substitutes. There were no complications. The ratio of screw to pedicle diameter was approximately 0.6, greater than in human studies. CONCLUSIONS: Despite differences between pedicle dimensions and screw angles, most screws could be inserted into the sheep pedicle using endoscopic or open techniques. Bone fusion was successful in both groups. Thus, minimally invasive, endoscopic transpedicular screw-plate fixation and posterolateral fusion is feasible, and as effective as the conventional open technique. PMID- 15363423 TI - Posterior lumbar interbody fusion using recombinant human bone morphogenetic protein type 2 with cylindrical interbody cages. AB - BACKGROUND CONTEXT: In a large series of human patients undergoing open anterior lumbar interbody fusion with a tapered titanium fusion cage, recombinant human bone morphogenetic protein type 2 (rhBMP-2) on an absorbable collagen sponge carrier has been shown to decrease operative time and blood loss, to promote osteoinduction and fusion and to be a safe and effective substitute for iliac crest harvesting. PURPOSE: The purpose of the study was to determine the clinical and radiographic outcomes in patients treated for single-level degenerative lumbar disc disease with a posterior interbody fusion, using stand-alone cylindrical threaded titanium fusion cages with either autogenous bone graft or rhBMP-2 and an absorbable collagen sponge carrier. STUDY DESIGN/SETTING: A prospective, randomized, nonblinded, 2-year pilot study at 14 investigational sites. PATIENT SAMPLE: Between March 1999 and December 1999, 67 patients with symptomatic, single-level degenerative lumbar disc disease of at least 6 months' duration underwent a single-level posterior lumbar interbody fusion using two paired cylindrical threaded titanium fusion devices. Patients were randomly assigned to one of two groups: one (n=34 patients) received rhBMP-2 on a collagen sponge carrier; the other (n=33 patients) autogenous iliac crest bone graft. OUTCOME MEASURES: Clinical outcomes were measured using low back and leg pain numerical rating scales, the Short Form (SF)-36, Oswestry Low Back Pain Disability Questionnaire and work status. Plain radiographs and computed tomographic scans were used to evaluate fusion at 6, 12 and 24 months after surgery. METHODS: In this prospective nonblinded study, 67 patients were randomly assigned to one of two groups who underwent interbody fusion using two cylindrical threaded fusion cages: the investigational group (34 patients), who received rhBMP-2 on an absorbable collagen sponge, and a control group (33 patients), who received autogenous iliac crest bone graft. Clinical data were collected and analyzed by a commercial entity. RESULTS: The mean operative time and blood loss for the investigational rhBMP-2 group was 2.6 hours and 322.8 ml, respectively. For the autograft control group, these values were 3.0 hours and 372.7 ml. The differences were not significant. Although not statistically different, at 24 months, the investigational group's fusion rate of 92.3% was higher than the control's at 77.8%. At all postoperative intervals, the mean Oswestry, back and leg pain scores and physical components of the SF-36 improved in both treatment groups compared with preoperative scores, but no significant differences were found between groups. A statistically significant difference in the change in back pain was found at 24 months for the investigational group. In the control group, two adverse events related to harvesting of the iliac crest graft occurred in two patients (6.1%). CONCLUSIONS: This small multicenter, randomized, nonblinded trial showed few statistically significant differences between the study groups. Both groups showed comparable improvements on outcome scores. Overall results show that the use of rhBMP-2 can eliminate the need for harvesting iliac crest graft and may be an equivalent replacement for autograft for use in successful posterior lumbar interbody fusions. Further studies of the use of rhBMP-2 in posterior lumbar interbody fusion cage procedures are needed. PMID- 15363426 TI - The endoscopically assisted simultaneous posteroanterior reconstruction of the thoracolumbar spine in prone position. AB - BACKGROUND CONTEXT: The reconstruction of the anterior column of the thoracolumbar spine has become more common in the last few years, due largely to the unfavorable results of exclusively posterior surgical treatment, which has been associated with a lack of about 10 degrees of kyphosis correction after removal of the instrumentation. The minimally invasive anterior techniques have reduced the morbidity of the anterior approach significantly. PURPOSE: A minimally invasive technique for anterior stabilization of the spine may reduce the morbidity of the open approach. Irrespective of an anterior open or an endoscopic approach, the posteroanterior instrumentation of thoracolumbar fractures requires time-consuming intraoperative maneuvers to change the patient position from prone to lateral. We describe here a standardized anterior endoscopically assisted approach for the segments T4 to L4. This approach allows the patient to remain in prone position. A 4- to 5-cm incision combined with a retractor system is used. STUDY DESIGN/SETTING: In a prospective study, all patients of our clinic who underwent surgery of the thoracolumbar spine between July 1999 and May 2001 were registered. Study criteria were duration of surgery, duration of anesthesia, intra- and postoperative complications. PATIENT SAMPLE: Between July 1999 and May 2001, 42 patients (25 male, 17 female, average age of 41.9 years), who presented with 55 injured spinal levels and underwent surgery of the thoracolumbar spine in prone position, were included. OUTCOME MEASURES: Duration of surgery (posterior/anterior/total), duration of anesthesia, method of instrumentation, intra- and postoperative complications, postoperative hospital stay and radiographs were evaluated. METHODS: Surgery was performed in prone position. A thoracic approach was used for instrumentation of T9 to L2. A retroperitoneal approach was used for stabilization of L1 to L5. Both procedures were endoscopically assisted with a new retractor system (Synframe; Synthes GmbH, Umkirch, Germany). In this manner, only an incision 4 to 5 cm long and a stab incision for the endoscope were required. The whole procedure was performed in prone position without a change of position during surgery. RESULTS: A total of 42 patients underwent surgery following this technique: 14 isolated anterior procedures (median duration of surgery, 181 minutes); 13 simultaneous one-stage procedures (median duration of surgery: 210 minutes) and 15 combined two-stage procedures (median duration of surgery: 90 minutes posterior, 120 minutes anterior, 240 minutes posterior+anterior). In the simultaneous posteroanterior procedures, the anterior instrumentation was performed 20 times using one rod, twice using two rods and in six patients simply by bone grafting. No intraoperative complications were observed. In the postoperative course, one case of pneumothorax, one case of hemothorax and one case of transient intercostal neuralgia occurred. CONCLUSION: The approach to the anterior spine in prone position is feasible by using a self-holding retractor system for the region between T4 and L4. The duration of anesthesia for the one-stage simultaneous procedure was reduced by about 40 minutes, because changing the position of the patient is no longer necessary. The minimal incision, in combination with the retractor system, significantly reduces cost by allowing the use of less expensive instruments and implants. The advantages of the open and the endoscopic techniques are combined, while their disadvantages are minimized. The main advantage of the prone position is the opportunity to access the anterior and posterior spine simultaneously, which is especially helpful in reduction maneuvers. PMID- 15363428 TI - The related outcome and complication rate in primary lumbar microscopic disc surgery depending on the surgeon's experience: comparative studies. AB - BACKGROUND CONTEXT: Studies concerning intraoperative complications and their influence on the clinical outcome of microscopic disc surgery are quite rare. Complication rates vary between 1.5% and 15.8%. A correlation between the surgeon's experience and the complication rate may be expected. PURPOSE: To determine the influence of the surgeon's experience on the intraoperative complication rate in lumbar microscopic disc surgery. STUDY DESIGN: Three studies are included: 1) retrospective analysis of intraoperative complications in microscopic disc surgery (N=1,872); 2) prospective follow-up study of microscopic disc surgery (N=583); 3) prospective evaluation of complication rates in microscopic disc surgery (N=90). PATIENT SAMPLE: Patient data sets from 1,872 lumbar microscopic disc surgeries performed between January 7, 1981, and June 31, 2000, were examined in a retrospective study. A total of 463 patients, operated on between 1991 and 1996, were followed up by a questionnaire. Finally, a prospective controlled trial (N=90) was performed. OUTCOME MEASURES: Such complications as incidental durotomy, wrong level exposure, or bleeding were analyzed based on the patient data sets by a blinded external evaluator. The rates of lower back pain and ischiatic pain were measured on a visual analogue scale at follow-up in Study 2 and Study 3. To measure the outcome of surgery in daily life activities and functional capacity, the Tegner activity level was calculated. In addition, a questionnaire with the Hannover score was used. The patient's social and economic status was also recorded. METHODS: A total of 1,872 lumbar microscopic disc surgeries, performed between January 7, 1981 and June 31, 2000, were examined in a retrospective study. Intra- and perioperative complications were evaluated and related to the surgeons' level of experience. Patients in the first group (XL) were operated on by the most experienced surgeon (more than 500 microscopic discectomies before the beginning of the study). The L group surgeons performed between 50 and 100 microscopic disc surgeries before the study. This group included a total number of seven surgeons during the 1981-2000 time frame. None of this group reached the experience level of 500 surgeries during the course of the study. A total of 463 patients, operated on between 1991 and 1996, were followed up. Finally, a prospective controlled trial (N=90) was performed. Injuries of the dura, nerve root, ventral structures and wrong level exposure, which had been detected and corrected during surgery, were analyzed. In the second and third study, the outcome was correlated to surgery and complications during surgery. RESULTS: The rate of intraoperative complications showed a statistically significant difference between the groups. The comparison of both groups (n=1,872) with regard to the rate of intraoperative complications showed a statistically significant difference between 2.2% in the XL group and 10.7% in the L group (p< or =.001). Regarding work-related and socioeconomic factors, no significant difference in the outcome was seen. CONCLUSIONS: Microscopic disc surgery requires a course of instruction and a considerable number of surgeries under supervision by experienced surgeons. To shorten the learning curve, a number of standardized surgery steps to clearly identify anatomical landmarks are helpful. During training, these landmarks can be checked by an experienced surgeon to minimize the rate of intraoperative complications. Initial postoperative ischiatic pain was correlated to an incidental durotomy with p<.001. For long-term results after disc surgery, however, socioeconomic and work-related factors are of greater importance in spinal disc surgery than the incidence of intraoperative complications. PMID- 15363429 TI - Procedural skills in spinal manipulation: do prerequisites matter? AB - BACKGROUND CONTEXT: Spinal manipulation has undergone a resurgence of interest. Developing evidence suggests a relationship between safety, skill and clinical outcome. Training programs are variable and range from extensive formalized curricula to weekend seminars and individual demonstrations. Systematic study of a relationship between prerequisites and skill development has not been conducted. PURPOSE: This project evaluated programmatic differences in prerequisites of students during their training for spinal manipulation with respect to quantitative biomechanical evidence of procedural control and skill of performance of a novel task. STUDY DESIGN/SETTING: The research used an experimental design comparing two cohorts involved in separate training programs at different institutions that had distinguishing characteristics in methods of prerequisites to manipulation training. METHODS: A common manipulation procedure (L4 mamillary push [L4MP]) was chosen as a standard test maneuver. Performance of the procedure on initial effort by two cohorts of students (n=38 vs n=39) entering into training for lumbar spine procedures was measured. Comparisons were made based on quantitative biomechanical parameters to assess control and skill. Results were compared with a cohort of experts as a reference standard. RESULTS: Significant differences were observed between the performance measures of the two cohorts. The more skilled performance group was more similar to the expert reference standard than was the lesser skilled group. CONCLUSIONS: The duration, extent and content of prerequisites for learning the dynamic and complex manual skills for spinal manipulation can significantly influence the level of skill attainment even early in the course of training. PMID- 15363430 TI - Posterolateral transforaminal selective endoscopic discectomy and thermal annuloplasty for chronic lumbar discogenic pain: a minimal access visualized intradiscal surgical procedure. AB - BACKGROUND CONTEXT: Chronic lumbar discogenic pain (CLDP) impairs the patient's physical abilities to function within the normal physiologic loading ranges of activities of daily living. The pathogenesis of CLDP is multifactorial and not well understood. Conservative therapeutic regimens often fail to achieve sufficient pain relief. Surgical options vary greatly in surgical invasiveness as well as outcome. Definitive surgical treatment is often 360-degree fusion. The morbidity associated with this approach is significant, considering that only 65% to 80% of patients obtain satisfactory clinical results. This has spawned interest in minimally invasive surgical options, such as intradiscal electrothermal therapy (IDET; ORATEC Interventions, Inc., Menlo Park, CA), but results are conflicting. PURPOSE: The authors describe their surgical technique of minimal access posterolateral transforaminal selective endoscopic discectomy (SED) and bipolar radiofrequency thermal annuloplasty to treat CLDP. The procedure's rationale is based on the hypothesis that annular defects are the focal points of chronic exposure between neural sensory receptors in the defect and the inflammatogenic nucleus pulposus. In contrast to other percutaneous procedures, this technique allows direct visualization and targeting of the disc nucleus and annular fissures. Our 2-year clinical result is reported. STUDY DESIGN/SETTING: This is a retrospective review of consecutive surgical cases performed by one surgeon (ATY). The procedures were carried out from January 1997 to December 1999. Each patient has a minimum postoperative follow-up of 2 years. PATIENT SAMPLE: A total of 113 patients met the generally accepted clinical criteria for chronic lumbar discogenic pain and were selected for the procedure. OUTCOME MEASURES: Two outcome measures were used for clinical assessment: a surgeon-based modified MacNab method and a patient-based questionnaire. A mandatory poor result was given to any patient who had repeat spine surgery at the same level or has indicated dissatisfaction with the surgical result on the questionnaire response. METHOD: After meeting CLDP selection criteria, provocation contrast/indigo carmine dye discography was performed. This test was used to confirm the suspected discs as pain generators. The subject surgery then followed. Only cases with one and two levels of confirmed painful discs were entered into the study. The nonoperating author (PMT) analyzed the data. RESULTS: Using the surgeon assessment method, 17 patients (15%) had excellent results, 32 patients (28.3%) had good results, 34 patients (30.1%) had fair results and 30 patients (26.5%) had poor results. Of the 30 patients in the poor result group, 12 reported either no improvement or worsening, and refused further surgical treatment. Of the remaining 18 patients in the poor group, 8 had spinal fusion, 3 had laminectomy and 7 had repeat spinal endoscopic surgery. The patient-based questionnaire yielded similar percentages in each category. However, only 73.5% of the 113 patients returned the survey questionnaire. There were no aborted procedures, unexpected hemorrhage, device-related complications, neurologic deficits, perioperative deaths or late instability. CONCLUSIONS: Posterolateral transforaminal SED and radiofrequency thermal annuloplasty were used to interrupt the purported annular defect pain sensitization process, thought to be necessary in the genesis of chronic lumbar discogenic pain. Lack of clinical benefit from the subject procedure did not degrade any subsequent surgical or nonsurgical treatment options. The experience gained from this study warrants further investigation into the cellular and molecular processes that provided back pain relief in these patients. PMID- 15363431 TI - Dose-response for chiropractic care of chronic low back pain. AB - BACKGROUND CONTEXT: There have been no trials of optimal chiropractic care in terms of number of office visits for spinal manipulation and other therapeutic modalities. PURPOSE: To conduct a pilot study to make preliminary identification of the effects of number of chiropractic treatment visits for manipulation with and without physical modalities (PM) on chronic low back pain and disability. STUDY DESIGN/SETTING: Randomized controlled trial with a balanced 4x2 factorial design. Conducted in the faculty practice of a chiropractic college outpatient clinic. PATIENT SAMPLE: Seventy-two patients with chronic, nonspecific low back pain of mechanical origin. MAIN OUTCOME MEASURES: Von Korff pain and disability (100-point) scales. METHODS: Patients were randomly allocated to visits (1, 2, 3 or 4 visits/week for 3 weeks) and to treatment regimen (spinal manipulation only or spinal manipulation with PM). All patients received high-velocity low amplitude spinal manipulation. Half received one or two of the following PM at each visit: soft tissue therapy, hot packs, electrotherapy or ultrasound. RESULTS: Pain intensity: At 4 weeks, there was a substantial linear effect of visits favoring a larger number of visits: 5.7 points per 3 visits (SE=2.3, p=.014). There was no effect of treatment regimen. At 12 weeks, the data suggested the potential for a similar effect of visits on patients receiving both manipulation and PM. Functional disability: At 4 weeks, a visits effect was noted (p=.018); the slope for group means was approximately 5 points per 3 visits. There were no group differences at 12 weeks. CONCLUSIONS: There was a positive, clinically important effect of the number of chiropractic treatments for chronic low back pain on pain intensity and disability at 4 weeks. Relief was substantial for patients receiving care 3 to 4 times per week for 3 weeks. PMID- 15363432 TI - Spinal injury considerations in the competitive diver: a case report and review of the literature. AB - BACKGROUND CONTEXT: Despite significant literature associated with spinal injuries and recreational diving, few articles exist regarding competitive diving injuries, with no reports pertaining specifically to spinal injuries. As a result, a case report of a collegiate diver with C5-C6 ligamentous instability requiring operative stabilization is currently presented in addition to a review of the literature. PURPOSE: Present a case report of cervical C5-C6 ligamentous instability in a collegiate diver. STUDY DESIGN: Case report and literature review. METHODS: Not applicable. RESULTS: Not applicable. CONCLUSIONS: Diving injuries pertaining to competitive diving do occur but to a lesser extent than would be expected given the large forces the spine experiences. Training, experience and appropriate technique greatly minimize potential spinal hazards. Persistent complaints of neck pain after a competitive diving injury mandate aggressive evaluation and further workup. PMID- 15363433 TI - Posterolateral odontoidectomy for irreducible atlantoaxial dislocation: a technical case report. AB - BACKGROUND CONTEXT: For the treatment of irreducible atlantoaxial dislocation (AAD), anterior decompression has been commonly used by removal of the odontoid process and fixation of the vertebral body by means of the transoral approach. Although the transoral approach has been well established, there are some disadvantages. By contrast, there have been few technical case reports of the posterolateral approach, especially for irreducible AAD. PURPOSE: To describe a technical case report of the posterolateral approach for irreducible AAD, demonstrating the specific advantages. STUDY DESIGN/SETTING: Technical case report. PATIENT SAMPLE: Case report. OUTCOME MEASURES: Report of postoperative symptoms. METHODS/DESCRIPTION: A 74-year-old woman presented with sudden left hemiparesis because of irreducible AAD. She underwent the odontoidectomy and reduction by means of the posterolateral approach. Posterior fixation with instrumentation and an iliac bone graft was performed 16 days later. The results of radiological examinations were satisfactory. The posterolateral approach takes advantage of the particular features of C1-C2 dislocation of irreducible AAD. CONCLUSIONS: We suggest the possibility that the posterolateral approach is feasible and effective in most cases with irreducible AAD. PMID- 15363434 TI - Acute spinal cord compression caused by vertebral hemangioma. AB - BACKGROUND CONTEXT: The reported incidence of vertebral hemangioma within the spinal column is common. Most often these benign vascular tumors are incidental radiographic findings and do not cause neurological sequelae. Rarely, vertebral hemangiomas will cause compressive neurological symptoms, such as radiculopathy, myelopathy and paralysis. In these cases the clinical presentation is usually the subacute or delayed onset of progressive neurological symptoms. This report demonstrates a symptomatic vertebral hemangioma presenting with rapid onset neurologic sequelae. PURPOSE: To discuss diagnostic and management issues presented by symptomatic vertebral hemangioma. STUDY DESIGN: Case report and review of literature. PATIENT SAMPLE: Sixty-one-year-old white woman with low back pain and rapidly progressive myelopathic symptoms. METHODS: A case of vertebral hemangioma with neurological sequelae is presented followed by a discussion of the literature concerning diagnostic and therapeutic options in the management of this pathologic entity. RESULTS: The results of our review reveal that the incidence of vertebral hemangioma causing compressive neurological symptoms is rare despite the overall prevalence of vertebral hemangioma. Vertebral hemangioma may present with rapid onset myelopathic symptoms and may mimic those symptoms caused by a malignancy. Radiographic imaging modalities are extremely useful and display characteristic findings in the diagnostic evaluation of these tumors. Angiographic embolization of feeding vessels has been effective in minimizing operative blood loss, and surgical decompression and stabilization is frequently indicated. Postsurgical radiotherapy has also been demonstrated to serve as a limited adjunct to surgery by reducing tumor recurrence in the event of less than complete tumor resection. CONCLUSIONS: Because of the rapid presentation of myelopathic symptoms in this case, preoperative angiographic embolization was not performed, and the patient underwent emergent decompressive surgery. In this case emergent operative decompression and stabilization was effective in reversing the patient's myelopathic symptoms, while maintaining long term stability of the spinal column. Postoperative radiation was not administered because of the extent of tumor resection. Surgical intervention has produced long term cure of this patient's myelopathy and T10 vertebral hemangioma. PMID- 15363435 TI - Predicting outcomes of transforaminal injections for sciatica. PMID- 15363437 TI - The changing landscape of antiparasitic drug discovery for veterinary medicine. AB - Changes in economic imperatives in the pharmaceutical industry have led to a wave of consolidation, which has had the unintended side effect of shrinking the resource devoted to antiparasitic drug discovery in animal health companies. Scientific changes have altered the way in which drugs could be discovered in the future. New science and business models will need to be implemented to address the demand for innovative antiparasitic drugs in veterinary medicine. Novel drugs are needed to combat drug resistance and for currently non-addressed problems. At the center of the future for this field, however, lies the need for more support into the basic research on the biology of parasites. PMID- 15363438 TI - Veterinary anthelmintics: old and new. AB - Between 1960 and 1980, extraordinary success was achieved in anthelmintic development for animals. In these 20 years, drugs with diverse structure, novel activity and enviable safety were produced for a global livestock industry leading to the productivity gains needed to support a human population that grew by 1.5 billion during the same period. The following 20 years have been spent refining existing molecules with niche activity (parasite and host specificity), improving delivery systems and worrying about the inexorable spread of drug resistance. The challenge for the next 20 years will be to use the technologies available to design and produce new drugs and biological controls. Then, to use the lessons of the past to ensure that the new drugs enjoy a longer useful lifespan and contribute to an animal health industry (livestock and companion) which enriches the lives of a global population. Old and new veterinary anthelmintics comprise a very large field, which could not be comprehensively reviewed in a short article. The present mini-review focuses on major chemical discoveries, formulation developments, administration strategies and new products. PMID- 15363439 TI - Genomic filtering: an approach to discovering novel antiparasitics. AB - Genomic filtering is a rapid approach to identifying and prioritizing molecular targets for drug discovery. For infectious disease applications, comparative genomics filters allow the selection of pathogen-specific gene products, whereas functional genomics filters, such as RNA interference (RNAi), allow the selection of gene products essential for pathogen survival. The approach is especially applicable to antiparasitic drug discovery where the phylogenetic distance between parasite and host make the likelihood of drug cross-toxicity due to conservation of molecular targets greater than for more distantly related pathogens such as prokaryotes. This article discusses some of the inherent challenges of applying genomics to the early steps of drug discovery and describes one successful comparative and functional genomics filtering strategy that has been implemented to prioritize molecular targets and identify chemical leads for nematode control. PMID- 15363440 TI - Drug resistance in veterinary helminths. AB - At present, there is no effective alternative to chemical control of parasitic helminths where livestock are grazed intensively. Resistance to anthelmintics has become a major problem in veterinary medicine, and threatens both agricultural income and animal welfare. The molecular and biochemical basis of this resistance is not well understood. The lack of reliable biological and molecular tests means that we are not able to follow the emergence and spread of resistance alleles and clinical resistance as well as we need. This review summarizes some of the recent findings on resistance mechanisms, puts forward some recommendations for limiting its impact and suggests some priorities for research in this area. PMID- 15363441 TI - Drug resistance in nematodes of veterinary importance: a status report. AB - Reports of drug resistance have been made in every livestock host and to every anthelmintic class. In some regions of world, the extremely high prevalence of multi-drug resistance (MDR) in nematodes of sheep and goats threatens the viability of small-ruminant industries. Resistance in nematodes of horses and cattle has not yet reached the levels seen in small ruminants, but evidence suggests that the problems of resistance, including MDR worms, are also increasing in these hosts. There is an urgent need to develop both novel non chemical approaches for parasite control and molecular assays capable of detecting resistant worms. PMID- 15363442 TI - Neuropeptide signalling: a repository of targets for novel endectocides? AB - The only available parasiticides with a spectrum of action that includes a broad range of helminth and arthropod parasites are the macrocyclic lactones. Designated endectocides, these drugs have action against both endoparasitic nematodes and ectoparasitic arthropods. Unfortunately, the discovery of such drugs is exceedingly rare and there is no evidence that novel endectocidal agents will be identified and developed in the short to medium term. However, the discovery of neuropeptides with motor-modulatory activities in both arthropods and helminths, coupled with recent progress in the characterization of invertebrate neuropeptide receptors, has the potential to propel neuropeptide signalling to the forefront of efforts to develop a novel endectocide. PMID- 15363443 TI - Veterinary parasitic vaccines: pitfalls and future directions. AB - Most available antiparasitic drugs are safe, cheap and highly effective against a broad spectrum of parasites. However, the alarming increase in the number of parasite species that are resistant to these drugs, the issue of residues in the food chain and the lack of new drugs stimulate development of alternative control methods in which vaccines would have a central role. Parasite vaccines are still rare, but there are encouraging signs that their number will increase in the next decade. The modern paradigm is that an understanding of parasite genes will lead to the identification of useful antigens, which can then be produced in recombinant systems developed as a result of the huge investment in biotechnology. However, we should also continue to devote efforts to basic research on the host-parasite interface. PMID- 15363444 TI - Nematode control in 'green' ruminant production systems. AB - Collectively, nematode parasites of domestic ruminants continue to pose the greatest disease problem in grazing livestock systems worldwide, despite the powerful and extensive chemotherapeutic arsenal available for their control. The widespread development of anthelmintic resistance, particularly in nematode parasites of small ruminants, and the trend towards non-chemical (ecological, organic, green) farming of livestock has provided an impetus for the research and development of alternative parasite control methods. This article provides a brief overview of the non-chemotherapeutic options for parasite control and how they might play a role either in organic farming or in other low-input farming systems. PMID- 15363445 TI - Involvement of inducible nitric oxide synthase in renal failure after mild hemorrhage. AB - The role of iNOS in rats after mild hemorrhaging was examined in this study. A mild hemorrhage (17% of total blood) induced a decrease of systemic blood pressure and heart rate, transiently followed by gradual recovery. The hemorrhage caused expression of renal iNOS mRNA and an increase in systemic NO products at 1 h after bleeding. Serum creatinine and serum urea nitrogen (UN) increased progressively up to 5 h after bleeding. Light microscopic findings showed that some inflammatory monocytes, mainly consisting of neutrophil, often existed in the glomerular capillaries, eosinophilic changes were observed in the cytoplasm at the proximal tubules, and urinary casts existed in the uriniferous space at 5 h after bleeding. The selective iNOS inhibitor, S-methylisothiourea (MTU), suppressed hemorrhagic expression of renal iNOS mRNA and systemic NO products, suppressed the increases of serum creatinine and UN, and improved renal histological aggravations induced by hemorrhaging. We speculated that MTU caused the negative circuit to suppress the renal failure through a decrease of NO generation. These results in the present study showed that iNOS expression induced by mild hemorrhaging at the early phase did participate in the development of renal failure. PMID- 15363446 TI - A new method for ABO genotyping using a multiplex single-base primer extension reaction and its application to forensic casework samples. AB - We developed a new method for ABO genotyping using a multiplex single-base primer extension reaction. The method allows for the simultaneous detection of six SNP sites in the ABO gene (nt 261, 297, 681, 703, 802, and 803) and the determination of ABO genotypes from their combinations. It enabled ABO genotyping of all samples of peripheral blood DNA extracted from 103 Japanese individuals, and had a highly satisfactory detection sensitivity being capable of genotyping 0.1 ng of genomic DNA. Using this method, we were able to determine ABO genotypes of minute stain samples, heated bloodstains, aged bloodstains and mixed samples. Experiments with samples from 26 animal species and bacterial samples to test the species-specificity of the method showed that genotyping was possible in the chimpanzee and gorilla, but their genotypes were extremely rare in humans. In addition, we applied this method to casework samples, and successfully determined ABO genotypes of bones, teeth, muscles, organs, nails, and semen-contaminated vaginal fluid in which ABO grouping by conventional serological techniques was not possible. This new method enables the sensitive, simultaneous detection of six SNP sites in the ABO gene by two specific reactions, i.e. PCR and a primer extension reaction. Therefore, it holds promise as an effective method of ABO genotyping particularly for forensic samples. PMID- 15363447 TI - Postmortem changes in tissue concentrations of triazolam and diazepam in rats. AB - The aim of this study was to investigate the postmortem changes of concentrations of triazolam and diazepam in the rat model and to confirm the factor causing the phenomenon. We administered triazolam or diazepam orally to rats and then sacrificed them 1 h after administration. Abdominal tissues including stomach content and small intestine, thigh muscle and heart blood were collected at 0, 12, 24 h after death, and postmortem changes of the two drug concentrations were compared. Drug concentrations in the samples were analyzed by a selected ion monitoring of gas chromatography/mass spectrometry. Gastrointestinal postmortem concentrations of triazolam and diazepam decreased. On the other hand, the drug concentrations in the liver and kidney increased markedly, and those in lung and heart blood increased slightly during postmortem periods by diffusion from the gastrointestinal tract. The patterns of both drug concentration changes were similar. However, the extent of the triazolam increase tended to be larger than that of diazepam. This difference may be accounted for by lower triazolam concentrations before death. The postmortem drug concentrations of thigh muscle did not change when administered with triazolam and diazepam. For both triazolam and diazepam, a significant correlation was observed in the drug concentration between the small intestine and the kidney at 24 h after death, indicating that major cause of the change in the drug concentration in the kidney was diffusion from the small intestine up to 24 h after death. The results in this study indicate that diazepam as well as triazolam diffuses from the gastrointestinal tract into the surrounding tissues after death and suggest that the diffusion is influenced by their pharmaceutical properties before death. PMID- 15363448 TI - Development of a screening method for the most commonly abused anticholinergic drugs in Jordan; trihexyphenidyl, procyclidine and biperiden. AB - A sensitive and rapid method for the simultaneous determination of three commonly abused anticholinergic drugs in Jordan; trihexyphenidyl, procyclidine, and biperiden in plasma and urine has been developed using solid phase extraction and GC-MS. Linearity was established from therapeutic to fatal concentrations of the three drugs; 5-300 ng/ml in plasma, with correlation coefficient r(2) > or = 0.9978 and 10-800 ng/ml in urine r(2) > or = 0.9993. Recoveries were in the range of 86-92% and intra-day and inter-day relative standard deviations (n = 6) were in the range of 6.6-10.3% for the three drugs at three different concentrations in plasma and urine. The base peak m/z 98 for trihexyphenidyl, m/z 84 for procyclidine, and m/z 98 and 218 for biperiden, and m/z 339 for papaverine (internal standard) were monitored at selective ion monitoring; their retention times were 8.10, 8.67 and 8.92 min, respectively, and 14.79 min for the internal standard with analysis time of 16.75 min. The limit of detection of 0.5 ng/ml was attained for trihexyphenidyl and procyclidine, while for biperiden 2.0 and 1.0 ng/ml in spiked plasma and urine, respectively. This method has been applied to forensic and authentic samples taken from abuser and patients using these drugs. The method will offer the clinicians and the legal authority the right diagnosis regarding the anticholinergic involved in any case of abuse with less than 1 h per sample (plasma or urine) from the time of receiving. PMID- 15363449 TI - A novel 56-bp variable tandem repeat polymorphism in the human deoxyribonuclease I gene and its population data. AB - This study confirms the presence of a novel variable number of tandem repeats polymorphism, designated as HumDN1, in intron 4 of the human deoxyribonuclease I (DNase I) gene. Genotyping was performed without difficulty by PCR-amplification and separation by agarose gel electrophoresis in 423 Japanese, originating from four geographically diverse areas in Japan, and 89 Germans. The HumDN1 allele variability was due to different numbers of 56-bp repeat sequences, and five different alleles were distinguished with apparent size between 364 and 588 bp. Although there was a general uniformity for the polymorphism in the Japanese population, significant differences in genotype distribution were found between the Japanese and German populations. Furthermore, linkage disequilibrium between the HumDN1 and DNase I protein polymorphisms was revealed. PMID- 15363450 TI - A simple method for domestic animal identification in Argentina using PCR-RFLP analysis of cytochrome b gene. AB - We developed a simple, quick assay in order to discriminate forensic samples among human, and common domestic and livestock species of the Pampean region, Argentina. A mitochondrial cytochrome b fragment amplified with universal primers was separately digested with three restriction enzymes (AluI, HaeIII, and HinfI) and the resulting fragments were resolved through electrophoresis in polyacrylamide gels. This PCR-RFLP method allowed us to identify the target species and worked on degraded samples. The assay was successfully applied in livestock robbery cases in Argentine, and may be useful when attempting a first assessment as to the specific status of a forensic evidence. PMID- 15363451 TI - An adult autopsy case of acute encephalopathy associated with influenza A virus. AB - On a cold winter morning, a 35-year-old male was unexpectedly found dead and therefore autopsied. Macro- and microscopically, the lungs were demonstrated bronchopneumonia. On the surface of brain, small blood vessels were slightly congested. Microscopically, brain edema was also observed, and proliferation of lymphocytes was observed around the capillary vessels of the hippocampus. These findings suggested a viral infection of the cerebrum. We conducted an immunohistochemical study with antibody against influenza virus. Influenza A virus antigen was detected in both the lungs and brain. Therefore, findings were compatible with influenza A encephalopathy. Even when serological inspection is not performed, it is useful to inspect localization of the virus antigen immunohistochemically. We considered that it is necessary to perform pathological examination for influenza encephalopathy in sudden death cases when influenza is epidemic. PMID- 15363452 TI - Acute subdural hematoma due to arteriovenous malformation primarily in dura mater: a case report. AB - We report a 42-year-old female with alcohol addiction who suddenly died of subdural hematoma (SDH) caused by dural arteriovenous malformation (AVM). In autopsy, there was seen a massive SDH with a total weight of 181 g that covered an entire part of the left cerebral hemisphere, although either serious external injuries of the head or any visible internal injuries of the brain were observed. SDH subsequently resulted in the tonsillar, transtentorial and subfalcial herniations with a right-sided shift of the left-lateral and third ventricles, and the left thalamus as well. Histopathological examination on the serial sections cut from the falx cerebri revealed abnormal distribution of arteries and veins with various sizes, which were comprehensively highlighted by immunohistochemical stainings with alpha-SMA and CD31. Although a very point of bleeding was not identified even by careful histological observation, we concluded that dural AVM could be critical for acute SDH in the present case. The value of ethanol concentration examined in the samples from SDH supported that the lesion could be not chronic, but acute. PMID- 15363453 TI - Postmortem painting of a cadaver. PMID- 15363454 TI - Application of the independent molecule model to the calculation of free energy and rigid-body motions of water heptamers and octamers. AB - Stabilization of clusters of water heptamer and octamer in the gas phase is studied with the independent molecule model. As indicators of stabilization, the H-bonding strength, the H-bonding lifetime and the free energy are calculated. For both the heptamers and octamers, clusters with one oxygen-ring and some branching H-bonded waters are more stable than clusters with multi-oxygen-rings. Both the heptamer and octamer prefer a shape intermediate between planar and polyhedral; the branched waters are highly fluctuating and consequently contribute to increasing the entropy of the cluster. As the temperature increases from 0 to 300 K, the entropy contributes to the differentiation in free energy of the cluster geometries. PMID- 15363455 TI - Modeling of p38 mitogen-activated protein kinase inhibitors using the Catalyst HypoGen and k-nearest neighbor QSAR methods. AB - We have employed in parallel the Catalyst HypoGen pharmacophore modeling approach and the variable selection k-nearest neighbor quantitative structure-activity relationship (kNN QSAR) method to model a diverse data set of p38 mitogen activated protein (MAP) kinase inhibitors. The HypoGen pharmacophore model, developed from a novel automated training set selection protocol, identified chemical functional features that were characteristic of the active compounds and differentiated the active from the inactive inhibitors. The kNN QSAR modeling employed topological descriptors and afforded predictive QSAR models with consistently high values of both leave-one-out cross-validated R2 for the training set and predictive R2 for the test set. The results of both modeling approaches were sensitive to the selection of the training and test sets used for model development and validation. The resulting Catalyst pharmacophore and kNN QSAR models can be used concurrently for rapid virtual screening of chemical databases to identify novel p38 MAP kinase inhibitors. PMID- 15363456 TI - Molecular basis of the constitutive activity and STI571 resistance of Asp816Val mutant KIT receptor tyrosine kinase. AB - The receptor tyrosine kinase, KIT, displays activating mutations in the kinase domain, which are associated with various cancers. We have used homology modelling based on the crystal structures of the insulin receptor kinase in active and inactive conformations to predict the corresponding structures of the KIT kinase domain. We have prepared four KIT models, one each for the active and inactive conformations of the wild-type and of the Asp816Val mutant proteins. We have also placed ATP into the active conformations and the inhibitor, STI571, into the inactive conformations. All models have been fully energy minimised. The molecular modelling studies described here explain (i) why Asp816Val KIT is constitutively active, (ii) why the nature of the substituting amino acid at residue 816 is relatively unimportant, and (iii) why the Asp816Val substitution confers resistance to the KIT-inhibitory drug STI571. The models will be valuable for predicting other kinase inhibitory drugs that may be active on wild-type and mutant forms of KIT. During the course of this work, a crystal structure of the active conformation of the KIT kinase domain has been published. Our model of the active conformation of the Asp816Val mutant is strikingly similar to this crystal structure, whereas our model of the active conformation of the wild-type kinase domain of KIT differs from the crystal structure in some respects. The reasons for this apparent discrepancy are discussed. PMID- 15363457 TI - Analysis of the activating mutations within the activation loop of leukemia targets Flt-3 and c-Kit based on protein homology modeling. AB - Molecular modeling provides a mechanistic hypothesis at the molecular level for the constitutive activation recently observed and reported for tyrosine protein kinases Flt-3 and c-Kit. Three-dimensional homology models for the active and inactive forms of these two kinases were made. Comparison of these models at the molecular level reveals that mutations of specific residues located in the activation loop (D835X and 836-deletion in Flt-3; D816V in c-Kit) as well as a 6 base pair (6-bp) insertion at residue 840 in Flt-3 operate in a similar way. Each mutation tends to weaken the forces that maintain the activation-loop folded inwards. None of the mutations are found to particularly stabilize the active state directly. The reason why the equilibrium is shifted towards the gate-open conformation of the protein is because, at least in these models, the mutations are found to critically destabilize the inactive conformational state of the kinase. PMID- 15363458 TI - Comparative estimation of vibrational entropy changes in proteins through normal modes analysis. AB - We compare the vibrational entropy changes of proteins calculated using a full and a number of approximate normal modes analysis methods. The vibrational entropy differences for three conformational changes and three protein binding interactions were computed. In general, the approximate methods yield good estimates of the vibrational entropy change in a fraction of the time required by full normal modes analysis. The absolute entropies are either overestimated or greatly underestimated, but the difference is sufficiently accurate for some methods. This indicates that some of the approximate methods can give reasonable estimates of the associated vibrational entropy changes, making them suitable for inclusion in free energy calculations. PMID- 15363459 TI - Computational analysis of tyrosine phosphatase inhibitor selectivity for the virulence factors YopH and SptP. AB - Bacterial pathogens such as Yersinia and Salmonella represent an important medical concern, causing human diseases ranging from gastrointestinal disease to the plague. The development of novel treatments of these bacterial infections has gained high priority recently due to the emergence of antibiotic resistance in these pathogens and the threat of the use of microbial agents as biological weapons. YopH of Yersinia and SptP of Salmonella are virulence factors that belong to the family of protein tyrosine phosphatases (PTPs). A great challenge remains in the design of selective PTPs inhibitors due to their highly conserved active site. In this paper, we present a comparative docking study to probe the selective inhibition of YopH and SptP with PTP1B in order to better understand their binding interactions with the bacterial tyrosine phosphates. Characterized binding sites in PTP1B were compared with YopH and SptP. Molecular dynamics simulations were used to incorporate ligand-induced conformational changes in the binding sites. These results, together with those binding modes and binding affinities distinguished in individual PTPs, provide insight into the structure based design of inhibitors for YopH and SptP. PMID- 15363460 TI - In silico activation of Src tyrosine kinase reveals the molecular basis for intramolecular autophosphorylation. AB - Structural data suggest that important hinge-bending motions of the two lobes that shape the catalytic domain of Src tyrosine kinase, together with reorganization of an alpha helix (helix C), are needed for the activation loop to adopt the catalytically competent conformation. The phosphorylation of a Tyr residue (Tyr-416) in this loop also seems to be essential for enzyme activation. However, no information is available about the dynamics of this activation process. By comparing the inactive and active forms of the catalytic domains of Src and Lck, another member of the Src family, we first identified a short stretch that can act as a hinge for the interlobe motion. The opening of the lobes was then simulated using a targeted molecular dynamics approach. The results obtained suggested that pulling the two lobes apart is not enough to induce the required conformational change in the activation loop. Rather unexpectedly, however, swinging of the lobes situated Tyr-416 in a suitable position for intramolecular autophosphorylation, and further simulation of Tyr 416-phosphorylated Src in the presence of ADP did then result in a conformational change that placed the activation loop in a position similar to that found in the active open conformation of Lck. Taken together, our results establish a physical link between intramolecular autophosphorylation and loop activation. PMID- 15363461 TI - Alignment of three-dimensional molecules using an image recognition algorithm. AB - This paper describes a novel approach, based on image recognition in two dimensions, for the atom-based alignment of two rigid molecules in three dimensions. The atoms are characterised by their partial charges and their positions relative to the remaining atoms in the molecule. Based on this information, a cost of matching a pair of atoms, one from each molecule, is assigned to all possible pairs. A preliminary set of intermolecular atom equivalences that minimises the total atom matching cost is then determined using an algorithm for solving the linear assignment problem. Several geometric heuristics are described that aim to reduce the number of atom equivalences that are inconsistent with the 3D structures. Those that remain are used to calculate an alignment transformation that achieves an optimal superposition of atoms that have a similar local geometry and partial charge. This alignment is then refined by calculating a new set of equivalences consisting of atom pairs that are approximately overlaid, irrespective of partial charge. A range of examples is provided to demonstrate the efficiency and effectiveness of the method. PMID- 15363462 TI - Variations in "rescuability" of immunoglobulin molecules from different forms of human lymphoma: implications for anti-idiotype vaccine development. AB - Idiotypic (Id) vaccination has shown promising results in patients with follicular lymphoma (FL). However, it still remains unclear whether the same approach might be suitable for the treatment of other B-cell malignancies. For this reason, we recently performed an interim analysis of patients proposed to receive this treatment at our center. The feasibility of employing idiotype vaccines was evaluated for five different B-cell malignancies in their first relapse, both in terms of induction and fusion, as well as overall treatment. Our data suggest that, unlike follicular lymphoma (87%), this approach is not feasible to treat other B-cell malignancies (0-20%) such as mantle cell, small lymphocytic, diffuse large cell and Burkitt's lymphoma (P < 0.01). The main difficulties encountered were technical problems related to the survival of idiotype-producing hybridomas (83%) and the early loss of idiotype production by growing hybridomas (17%). However, it remains possible that an idiotype vaccine might still be produced through molecular means for most, if not all cases of relapsing B-cell malignancies. PMID- 15363463 TI - Protein tyrosine phosphatases and breast cancer. AB - Protein tyrosine phosphatases (PTPs) consist of a large family of related enzymes, including the group of classical PTPs with its two main subgroups, the transmembrane receptor-type (RPTPs) and the intracellular or non-transmembrane PTPs. Published data on the expression and function of a panel of these enzymes in normal and cancerous breast tissues are discussed in this review. While most studies, albeit on different enzymes, have tended to agree on the evidence for an increased PTP expression in breast cancer, any connection between PTP expression and the enzymes' role in cancer development and progression remains largely open to interpretation. Concomitant increases of protein tyrosine kinase (PTK) and PTP activities in many cancers further indicate that a complex dysregulation in the balance of tyrosine phosphorylation could be responsible for major alterations in various cellular processes controlling tissue homeostasis. In particular, any relationship between the expression of PTPs and their specific diverse roles in the regulation of cell growth and apoptosis in breast cancer needs to be addressed in major fundamental, preclinical and clinical studies. PMID- 15363464 TI - FDG-PET in the clinical management of Hodgkin lymphoma. AB - Positron emission tomography (PET) is a molecular functional imaging technique that provides qualitative and quantitative information about the localization and activity of pathophysiological processes. The most commonly used tracer for oncological purposes is 2-[18F]fluoro-2-deoxy-d-glucose (FDG). FDG-PET has within recent years become the most important nuclear medicine imaging modality in the management of lymphoma. This review summarizes the data published so far concerning the value of FDG-PET in staging, treatment monitoring, therapy planning, and follow-up of Hodgkin lymphoma (HL). FDG-PET detects more disease sites and involved organs than conventional staging procedures including computerized tomography (CT) and has a large influence on staging. FDG-PET during and after therapy appears to provide considerable prognostic information. However, the impact on patient outcome is not clear since no controlled trials are conducted and follow-up periods are generally short. The value of dual modality PET/CT and its potential role in the radiotherapy planning is discussed. PMID- 15363465 TI - The role of chemotherapy in locally advanced, metastatic and recurrent cervical cancer. AB - Cervical cancer is among the major health problems world-wide although advances in screening programs. Surgery and radiotherapy are the treatment modalities of choice for early and locally advanced cervical cancer. However, the role of chemotherapy in this setting has been better investigated in the latest years. To improve loco-regional control in locally advanced disease, authors have tested both neo-adjuvant chemotherapy and concurrent chemoradiotherapy. From 1999 NCI clinical announcement, concurrent cisplatin-based chemoradiation is considered the treatment of choice for cervical cancer patients requiring radiation therapy. Neo-adjuvant chemotherapy is reaching encouraging results in IB bulky-IIA cervical cancer, but further investigation are ongoing in locally advanced cervical setting. The optimal treatment for patients with metastatic or recurrent cervical cancer is still undefined and chemotherapy is used with palliation intent. Cisplatin remains the most active cytotoxic agents, although combinations of cisplatin with paclitaxel, topotecan, vinorelbine, have shown encouraging results in phase II and in early phase III studies. This paper reviews the role of chemotherapy in the management of patients with locally advanced, metastatic and recurrent cervical cancer. Studies discussed in this paper were selected trough a search in the med-line database performed in October 2003. PMID- 15363466 TI - Predictive factors for response to chemotherapy in colorectal cancer patients. AB - Colorectal cancer represents a major health problem in the western world. A lot of drugs have been employed in treatment of this disease, but only few data are available about predictive factors for response to anticancer treatments in colorectal cancer. Aim of this paper is to review the main data about this investigation field. Using a Medline database search (1966-2003) we reviewed all the relevant papers that investigate clinical and molecular predictors for response to the main drugs used in the treatment of colorectal cancer patients, both in adjuvant and in advanced setting. Moreover we comprehensively reviewed all the data published in abstract form during the most significant international meetings. Our review put in evidence the most important predictive factors for response in colorectal cancer patients treated with anticancer chemotherapy both in adjuvant and in advanced setting. The predictive factors are clustered on the basis of the different anticancer drugs. The results of this review provide the rationale basis for personalizing anticancer treatment in colorectal cancer patients by molecular and clinical features, aiming to improve response rate and reduce toxicities. PMID- 15363467 TI - Neoadjuvant treatment for locally advanced rectal carcinoma. AB - Rectal cancer is one of the most common neoplasms of Western Countries. Overall mortality at 5 years is about 40%. This cancer is commonly diagnosed at a precocious stage, but because of local relapse and/or metastatic disease, only half of radically resected patients can be considered disease free. The value of adding radiotherapy to surgery in the treatment of patients with resectable rectal cancer has been assessed in trials using either preoperative or postoperative irradiation. Preoperative irradiation is more "dose-effective" than postoperative radiotherapy; that is, a higher dose is needed postoperatively to reduce rates of local recurrence to the same extent as preoperative radiation. Nevertheless, preoperative treatment has not been routinely recommended, mainly because it has not been shown to improve overall survival and because in some trials it has been associated with increased postoperative mortality. This paper critically reviews clinical trials of chemoradiotherapy on whether an optimal combination exists for locally advanced rectal cancer. Even if in the latest years, recent advances in surgery have improved the local control of disease, the next steps in rectal cancer care should aim at the improvement of local cure rates and the enhancement of systemic control. New approaches to CT treatment are necessary. Patient enrollment into rigorous and well-conducted clinical trials will generate new information regarding investigational therapies and it will offer improved therapies for patients with this disease. PMID- 15363468 TI - The role of uracil-tegafur (UFT) in elderly patients with colorectal cancer. AB - 5-Fluorouracil (5-FU) administered in several schedules since its introduction in 1957 continues to be an integral part of standard first-line therapy for colorectal cancer. Continuous intravenous (i.v.) infusion appears to yield improved response rate and overall survival, with fewer adverse effects compared with i.v. bolus dosing. However, these protracted infusions require portable infusion pumps and central venous lines, which are associated with complications (i.e. increased risk of infection and clotting and/or dislodgement of the catheter, increased risk of venous thrombosis). Colorectal carcinoma is the second cause of death for tumour after lung cancer. About 70% of cases occur over 65 years and 50% or more affects people over 70. In clinical research age was a common exclusion criteria and little information is available about the efficacy, safety and toxicity of chemotherapy in elderly patients because few studies focused on the treatment of cancer in that part of population. The goal of this article is to review the literature concerning the treatment of elderly patients with UFT, an orally administered dihydropyrimidine dehydrogenase (DPD) inhibitory fluoropyrimidine. PMID- 15363470 TI - Neurotrophic factors and the pathophysiology of schizophrenic psychoses. AB - The aim of this review is to summarize the present state of findings on altered neurotrophic factor levels in schizophrenic psychoses, on variations in genes coding for neurotrophic factors, and on the effect of antipsychotic drugs on the expression level of neurotrophic factors. This is a conceptual paper that aims to establish the link between the neuromaldevelopment theory of schizophrenia and neurotrophic factors. An extensive literature review has been done using the Pub Med database, a service of the National Library of Medicine, which includes over 14 million citations for biomedical articles back to the 1950s. The majority of studies discussed in this review support the notion of alterations of neurotrophic factors at the protein and gene level, respectively, and support the hypothesis that these alterations could, at least partially, explain some of the morphological, cytoarchitectural and neurobiochemical abnormalities found in the brain of schizophrenic patients. However, the results are not always conclusive and the clinical significance of these alterations is not fully understood. It is, thus, important to further neurotrophic factor research in order to better understand the etiopathogenesis of schizophrenic psychoses and, thus, potentially develop new treatment strategies urgently needed for patients suffering from these devastating disorders. PMID- 15363471 TI - Psychosis, migration and social environment: an age--and--gender controlled study. AB - PURPOSE: Adverse social experiences are frequently invoked to explain the higher rate of psychosis among migrant groups. The aim of the present study was to establish the socio-environmental factors distinguishing migrant psychotic patients from autochthonous patients. SUBJECTS AND METHOD: We conducted a cross sectional survey involving 341 migrant psychotic patients matched for age and gender with 341 autochthonous psychotic patients. RESULTS: Migrant patients lived more often with their parental family, were less often enrolled with a referral psychiatrist, presented a lower rate of employment, a lower percentage of alcohol misuse and of suicide attempts. DISCUSSION: Our findings add to the growing body of results showing that more attention needs to be focused on socio-environmental variables in psychosis research. However, several limitations have to be taken into account, particularly with regard to selection biases and age of onset of the psychotic illness. CONCLUSION: Our results are compatible with the hypothesis that unemployment is a contributing factor in the risk for psychosis among migrant groups. Migrants' families are an important keystone in the mental health care process of their sick relatives. Our service models need to be adapted with the aim to make the treatment easier for migrant patients. PMID- 15363472 TI - Analogical reasoning in schizophrenic delusions. AB - PURPOSE: Reasoning ability has often been argued to be impaired in people with schizophrenic delusions, although evidence for this is far from convincing. This experiment examined the analogical reasoning abilities of several groups of patients, including non-deluded and deluded schizophrenics, to test the hypothesis that performance by the deluded schizophrenic group would be impaired. SUBJECTS/MATERIALS: Eleven deluded schizophrenics, 10 depressed subjects, seven non-deluded schizophrenics and 16 matched non-psychiatric controls, who were matched on a number of key variables, were asked to solve an analogical reasoning task. RESULTS: Performance by the deluded schizophrenic group was certainly impaired when compared with the depressed and non-psychiatric control groups though less convincingly so when compared with the non-deluded schizophrenic group. The impairment shown by the deluded schizophrenic group seemed to occur at the initial stage of the reasoning task. DISCUSSION: The particular type of impairment shown by the deluded subjects was assessed in relation to other cognitive problems already researched and the implications of these problems on reasoning tasks and theories of delusions was discussed. PMID- 15363473 TI - A possible association between the CCK-AR gene and persistent auditory hallucinations in schizophrenia. AB - Recent studies have suggested that DNA variations in the CCK-AR gene might predispose individuals to schizophrenia and particularly to auditory hallucinations (AH). The aim of this study is to assess the association between AH, using a specific scale for AH in schizophrenia (PSYRATS), and the CCK-AR polymorphism at 779 in a Spanish sample. A total of 105 DSM-IV schizophrenic patients with AH and 93 unrelated controls were studied. Twenty-two patients were considered as persistent auditory hallucinators, which showed similar clinical and demographic characteristic than patients with episodic AH, but with the exception of the PSYRATS values. The persistent AH group showed an excess of the A1 allele when was compared with episodic or control groups. Our data support the possible role of the CCK-AR gene in the development of persistent AH in schizophrenic patients. PMID- 15363474 TI - The CLDN5 locus may be involved in the vulnerability to schizophrenia. AB - The present study was designed to detect three single nucleotide polymorphisms (SNPs) located on 22q11 that was thought as being of particularly importance for genetic research into schizophrenia. We recruited a total of 176 Chinese family trios of Han descent, consisting of mothers, fathers and affected offspring with schizophrenia for the genetic analysis. The transmission disequilibrium test (TDT) showed that of three SNPs, rs10314 in the 3'-untranslated region of the CLDN5 locus was associated with schizophrenia (chi(2) = 4.75, P = 0.029). The other two SNPs, rs1548359 present in the CDC45L locus centromeric of rs10314 and rs739371 in the 5'-flanking region of the CLDN5 locus, did not show such an association. The global chi-square (chi(2)) test showed that the 3-SNP haplotype system was not associated with schizophrenia although the 1-df test for individual haplotypes showed that the rs1548359(C)-rs10314(G)-rs739371(C) haplotype was excessively non-transmitted (chi(2) = 5.32, P = 0.02). Because the claudin proteins are a major component for barrier-forming tight junctions that could play a crucial role in response to changing natural, physiological and pathological conditions, the CLDN5 association with schizophrenia may be an important clue leading to look into a meeting point of genetic and environmental factors. PMID- 15363475 TI - Safety and tolerability of olanzapine compared with other antipsychotics in the treatment of elderly patients with schizophrenia: a naturalistic study. AB - OBJECTIVE: To evaluate the safety and tolerability of olanzapine in the treatment of elderly patients with schizophrenia. METHODS: A total of 135 outpatients with schizophrenia > or =60 years of age were treated with olanzapine (n = 105) or another antipsychotic (n = 30) and followed up for 6 months. Safety measures included the recording of spontaneous adverse events and extrapyramidal symptoms (EPS). Clinical status and effectiveness of the medications were measured using the Clinical Global Impressions-Severity of Illness and the Global Assessment of Function (GAF) scales. Quality of life was assessed by means of the Spanish version of the EuroQol. The Awad scale was applied to evaluate patients' subjective attitude towards medication. RESULTS: The incidence of overall adverse events and EPS was non-significantly lower in patients treated with olanzapine than in patients treated with other antipsychotics. The use of anticholinergic drugs was significantly lower (P = 0.04) in patients treated with olanzapine. Both groups of patients experienced similar improvements in Clinical Global Impressions-Severity and GAF scores. Non-significantly greater improvement in the acceptance of medication occurred at endpoint in olanzapine-treated patients than in control patients as measured by the Awad scale. The improvement in the EuroQol quality of life scale achieved at the end of study did not differ between both treatment groups. CONCLUSIONS: Results from this naturalistic study showed that olanzapine was as safe and effective as other antipsychotic drugs in the treatment of elderly patients with schizophrenia. PMID- 15363476 TI - Adjunctive use of reboxetine in schizophrenia. AB - BACKGROUND: Schizophrenia is frequently complicated by depressive or negative symptoms that respond only moderately to treatment with antipsychotic drugs. Reboxetine is a novel antidepressant, which inhibits the reuptake of norepinephrine. We sought to study the efficacy and tolerability of the adjunctive use of reboxetine in a cohort of schizophrenic patients with prominent depressive or negative symptoms. METHODS: Sixteen schizophrenic inpatients were recruited for this study. All subjects received 4-8 mg of reboxetine/day while the antipsychotic medication (typical antipsychotics = 4; atypical antipsychotics = 12) was continued. All subjects underwent a standardized assessment including PANSS, CGI, HAMD, and CDSS before and after treatment with reboxetine (mean 26 +/ 17 d). RESULTS: All subjects tolerated treatment with reboxetine. Adverse effects were mild and did not require discontinuation of reboxetine. All clinical scores (PANSS 93.1 vs. 63.1; CGI 5.4 vs. 4.1; HAMD 20.4 vs. 8.1; CDSS 12.5 vs. 4.6) improved significantly under adjunctive treatment with reboxetine (all P < 0.01). CONCLUSION: The adjunctive use of reboxetine in schizophrenic patients was safe and well-tolerated. Our results suggest that the adjunctive use of reboxetine may be an effective treatment for depressive and negative symptoms in schizophrenia. PMID- 15363477 TI - Visual-spatial processing and dimensions of schizophrenia: a preliminary study on 62 schizophrenic subjects. AB - The aim of the study was to explore the relationships between the performance in figure-ground segregation and the clinical dimensions in schizophrenia. Sixty-two schizophrenic subjects filled out the embedded figures test (EFT) and the schizophrenic symptomatology was rated using the brief psychiatric rating scale (BPRS). The BPRS items were grouped into four categories: positive, negative, general and disorganization. The results showed in male schizophrenics negative and significant correlations between EFT scores and negative or disorganization subscales. Taken into account that in schizotypic subjects EFT scores was negatively correlated with negative dimension of schizotypy we suggested that those relationship could constitute a trait of the disease. PMID- 15363478 TI - No association between polymorphisms in three genes of cytochrome p450 family and paranoid schizophrenia in northern Chinese Han population. AB - We genotyped six SNPs in the genes of p450 family among paranoid schizophrenics and normal controls. All subjects are unrelated Han Chinese. Three showed polymorphic, and no significant differences in allele or genotype frequencies were detected between patients and controls. Thus we obtained no evidence for the involvement of the polymorphisms in paranoid schizophrenia in the population investigated. PMID- 15363479 TI - An association study between polymorphisms in three genes of 14-3-3 (tyrosine 3 monooxygenase/tryptophan 5-monooxygenase activation protein) family and paranoid schizophrenia in northern Chinese population. AB - We performed an association study between three SNPs in the genes of 14-3-3 family and paranoid schizophrenia. SNP rs983583 G/A in the YWHAZ gene showed significant association with paranoid schizophrenia. Our study indicated that the YWHAZ gene was a potential susceptibility gene for paranoid schizophrenia in the population studied. PMID- 15363480 TI - Combined therapy with low-potency neuroleptic levomepromazine as an adjunct to haloperidol for agitated patients with acute exacerbation of schizophrenia. PMID- 15363481 TI - Repetitive transcranial magnetic stimulation does not potentiate antidepressant treatment. AB - In a double blind controlled study, rTMS results in a similar antidepressant effect to sham in combination with paroxetine. Both groups had the same delay in scale's scores improvement. rTMS seems not to be efficient as an add-on treatment to pharmacological medication in non-resistant major depression. PMID- 15363482 TI - Normalization of impaired cognitive functions failed to improve clinical symptomatology in a schizophrenic patient. PMID- 15363483 TI - Treatment of a rare psychopathological phenomenon: tactile hallucinations and the delusional other. PMID- 15363484 TI - Blepharospasm associated with olanzapine: a case report. PMID- 15363485 TI - Tardive dyskinesia following brief exposure to risperidone--a case study. PMID- 15363486 TI - New insights in developmental origins of different GnRH (gonadotrophin-releasing hormone) systems in perciform fish: an immunohistochemical study in the European sea bass (Dicentrarchus labrax). AB - The knowledge of the roles and origins of different gonadotrophin-releasing hormone (GnRH) systems could greatly contribute to improve the understanding of mechanisms involved in the physiological control of early development, puberty and spawning. Thus, in this study, we have analyzed the distribution of the cells expressing salmon GnRH, seabream GnRH and chicken GnRH-II forms in the brain and pituitary of developing sea bass using specific antibodies to their corresponding GnRH-associated peptides. The first prepro-chicken GnRH-II-immunoreactive cells arose in the germinal zone of the third ventricle at 4 days after hatching, increasing their number from days 10 to 30, in which they adopted their adult position. The prepro-chicken GnRH-II-immunoreactive fibers became conspicuous in the first week and from day 26 they reached almost all brain areas, especially the hindbrain, being never detected in the pituitary. First prepro-salmon GnRH immunoreactive cells were detected in the olfactory placode at day 7 after hatching and reached the olfactory bulbs at day 10. Migrating prepro-salmon GnRH cells arrived at the ventral telencephalon at day 15, and became apparent in the preoptic area from day 45. The prepro-salmon GnRH innervation was more evident in the forebrain and increased notably between 10 and 30 days, at which fibers already extended from the olfactory bulbs to the medulla. A few prepro-salmon GnRH-immunoreactive fibers were observed in the pituitary from day 30. The prepro seabream GnRH-immunoreactive cells were first detected at day 26 in the rostral olfactory bulbs. On day 30, prepro-seabream GnRH-immunoreactive cells were also present in the ventral telencephalon, reaching the preoptic area and the hypothalamus at 45 and 60 days, respectively. The prepro-seabream GnRH innervation appeared restricted to the ventral forebrain, increasing notably during the sixth week, when fibers also reached the pituitary. A significant prepro-seabream GnRH innervation was not detected in the pituitary until day 60. PMID- 15363487 TI - Subregional analysis of GABA(A) receptor subunit mRNAs in the hippocampus of older persons with and without cognitive impairment. AB - We employed in situ hybridization and quantitative densitometry techniques to examine hippocampal mRNA expression of GABA(A) receptor subunits alpha1 and alpha5 in human subjects with progressing cognitive impairment. Included in this study were 17 participants of the Religious Order Study (ROS), who were categorized into three groups based upon degree of cognitive impairment: no cognitive impairment (n = 6); moderate cognitive impairment (n = 5); and probable Alzheimer's disease (AD) (n = 6). While the levels of each specific subunit mRNA were relatively homogeneously distributed throughout the five hippocampal subregions analyzed (CA1-4, and the granule cell layer of the dentate gyrus), mRNA expression of the alpha1 receptor subunit was found to be 20% reduced in the moderate cognitive impairment group as compared to the no cognitive impairment group. In addition, alpha1 mRNA expression was 25% reduced in the probable Alzheimer's disease group compared to the group with no cognitive impairment. Similarly, alpha5 subunit mRNA was reduced 32% between no cognitive impairment and moderate cognitive impairment groups, and 35% between no cognitive impairment and probable Alzheimer's disease groups. No significant reductions were found between moderate cognitive impairment and probable Alzheimer's disease groups for either subunit. Collectively, our data provide evidence for modest reductions in GABA(A) receptor subunit mRNAs, and suggest these changes occur very early in the progression of Alzheimer's disease cognitive impairment. PMID- 15363488 TI - Abundance and location of DARPP-32 in striato-tegmental circuits of domestic chicks. AB - The striatum is reciprocally connected to the brainstem dopaminergic nuclei and receives a strong dopaminergic input. In the present study the spatial relation between the dopaminergic and dopaminoceptive structures of the avian medial striatum (formerly: lobus parolfactorius) was observed by confocal laser scanning microscope in the domestic chick (Gallus domesticus). We also analysed the connections in the area ventralis tegmentalis and the substantia nigra. To label the dopaminergic structures, anti-tyrosine hydroxylase was used and DARPP-32 (dopamine and cAMP regulated phosphoprotein) was a marker of dopaminoceptive elements. The tyrosine hydroxylase positive fibres formed baskets of juxtapositions around the DARPP-32 containing cells of the medial striatum. However, such baskets were also observed to juxtapose DARPP-32 immunonegative cells. In the tegmentum, DARPP-32 was observed in axons descending from the telencephalon via the ansa lenticularis. These varicose fibers innervated the ventral tegmental area and substantia nigra and were often juxtaposed to dopaminergic neurons and dendrites. Approximately 40% of the striatal projection neurons targeting the ventral tegmentum, and 60% of striatal projection neurons targeting the nigra were immunoreactive to DARPP-32, as revealed by retrograde pathway tracing with Fast Blue. Endogenous dopamine may exert a retrograde synaptic effect on the afferent striato-tegmental fibers, apart from the reported extrasynaptic action. The abundance of juxtapositions observed in the avian brainstem and medial striatum corroborates the possibility of reciprocal striato tegmental circuits, relevant to the reinforcement of behaviour. PMID- 15363489 TI - Extrabulbar olfactory system and nervus terminalis FMRFamide immunoreactive components in Xenopus laevis ontogenesis. AB - The extrabulbar olfactory system (EBOS) is a collection of nerve fibers which originate from primary olfactory receptor-like neurons and penetrate into the brain bypassing the olfactory bulbs. Our description is based upon the application of two neuronal tracers (biocytin, carbocyanine DiI) in the olfactory sac, at the cut end of the olfactory nerve and in the telencephalon of the developing clawed frog. The extrabulbar olfactory system was observed already at stage 45, which is the first developmental stage compatible with our techniques; at this stage, the extrabulbar olfactory system fibers terminated diffusely in the preoptic area. A little later in development, i.e. at stage 50, the extrabulbar olfactory system was maximally developed, extending as far caudally as the rhombencephalon. In the metamorphosing specimens, the extrabulbar olfactory system appeared reduced in extension; caudally, the fiber terminals did not extend beyond the diencephalon. While a substantial overlapping of biocytin/FMRFamide immunoreactivity was observed along the olfactory pathways as well as in the telencephalon, FMRFamide immunoreactivity was never observed to be colocalized in the same cellular or fiber components visualized by tracer molecules. The question whether the extrabulbar olfactory system and the nervus terminalis (NT) are separate anatomical entities or represent an integrated system is discussed. PMID- 15363491 TI - ER chaperone functions during normal and stress conditions. AB - Nearly all resident proteins of the organelles along the secretory pathway, as well as proteins that are expressed at the cell surface or secreted from the cell, are first co-translationally translocated into the lumen of the endoplasmic reticulum (ER) as unfolded polypeptide chains. Immediately after entering the ER, they are often modified with N-linked glycans, are folded into the appropriate secondary and tertiary structures, which are stabilized by disulfide bonds, and finally in many cases are assembled into multimeric complexes. These processes are aided and monitored by ER chaperones and folding enzymes. When cells experience conditions that alter the ER environment, protein folding can be dramatically affected and can lead to the accumulation of unfolded proteins in this organelle. This in turn activates a signaling response, which is shared among all eukaryotic organisms, termed the unfolded protein response (UPR). The hallmark of this response is the coordinate transcriptional up-regulation of ER chaperones and folding enzymes. A major role for the increased levels of chaperones and folding enzymes during conditions of ER stress is to provide the same functions they carry out during normal physiological conditions. This includes preventing unfolded and incompletely folded proteins from aggregating and promoting the proper folding and assembly of proteins in the ER. During conditions of ER stress, many proteins are unable to fold properly and the requirements for chaperones are therefore increased. However, more recently it has become clear that some ER chaperones are also involved in signaling the ER stress response, targeting misfolded proteins for degradation and perhaps even shutting down the UPR when the stress subsides. In addition, during some normal physiological conditions, like plasma cell differentiation where there is an increased demand in the secretory capacity of B cells, the levels of various ER chaperones are also up-regulated via at least part of the UPR pathway. In order to discuss these various functions of ER chaperones, we will begin with the roles of ER chaperones and folding enzymes during normal physiological conditions and then discuss their roles during ER stress. PMID- 15363492 TI - Induction of neuronal death by ER stress in Alzheimer's disease. AB - Recent studies have suggested that neuronal death in Alzheimer's disease (AD) or ischemia could arise from dysfunction of the endoplasmic reticulum (ER). Inhibition of protein glycosylation, perturbation of calcium homeostasis, and reduction of disulfide bonds provoke accumulation of unfolded protein in the ER, and are called 'ER stress'. Normal cells respond to ER stress by increasing transcription of genes encoding ER-resident chaperones such as GRP78/BiP, to facilitate protein folding or by suppressing the mRNA translation to synthesize proteins. These systems are termed the unfolded protein response (UPR). Familial Alzheimer's disease-linked presenilin-1 (PS1) mutation downregulates the unfolded protein response and leads to vulnerability to ER stress. The mechanisms by which mutant PS1 affects the ER stress response are attributed to the inhibited activation of ER stress transducers such as IRE1, PERK and ATF6. On the other hand, in sporadic Alzheimer's disease (sAD), we found the aberrant splicing isoform (PS2V), generated by exon 5 skipping of the Presenilin-2 (PS2) gene transcript, responsible for induction of high mobility group A1a protein (HMGA1a). The PS2V also downregulates the signaling pathway of the UPR, in a similar fashion to that reported for mutants of PS1 linked to familial AD. It was clarified what molecules related to cell death are activated in the case of AD and we discovered that caspase-4 plays a key role in ER stress-induced apoptosis. Caspase-4 also seems to act upstream of the beta-amyloid-induced ER stress pathway, suggesting that activation of caspase-4 might mediate neuronal cell death in AD. PMID- 15363493 TI - The unfolded protein response--a stress signaling pathway of the endoplasmic reticulum. AB - The endoplasmic reticulum (ER) is a factory for folding and maturation of newly synthesized transmembrane and secretory proteins. The ER provides stringent quality control systems to ensure that only correctly folded proteins exit the ER and unfolded or misfolded proteins are retained and ultimately degraded. A number of biochemical and physiological stimuli can change ER homeostasis, impose stress to the ER, and subsequently lead to accumulation of unfolded or misfolded proteins in the ER lumen. The ER has evolved stress response signaling pathways collectively called the unfolded protein response (UPR) to cope with the accumulation of unfolded or misfolded proteins. This review summarizes our understanding of the UPR signaling developed in the recent years. PMID- 15363494 TI - Survival and apoptosis signals in ER stress: the role of protein kinases. AB - The endoplasmic reticulum (ER) is the organelle in which newly synthesized secretory and transmembrane proteins form their proper tertiary structure by post translational modification, folding, and oligomerization. However, many of these proteins are unfolded or misfolded by extracellular or intracellular stimuli. The accumulation of misfolded proteins constitutes a risk for living cells. Eukaryotic cells possess at least three different mechanisms to adapt to ER stress and thereby survive: (1) translational attenuation to limit further accumulation of misfolded proteins; (2) transcriptional activation of genes encoding ER-resident chaperones; and (3) the ER-associated degradation (ERAD) pathway to restore the folding capacity. If the cells are exposed to prolonged or strong ER stress, the cells are destroyed by apoptosis. Recent evidence indicates that ER stress signaling pathways are mediated in part by several protein kinases and play an important role in the pathogenesis of neurodegenerative disorders. The main purpose of this review is to summarize current knowledge about the protein kinases involved in ER stress, and their involvement in the pathogenesis of neurodegenerative disorders. PMID- 15363495 TI - Caspases involved in ER stress-mediated cell death. AB - Caspases are cysteine proteases involved in apoptotic pathways. Excess endoplasmic reticulum (ER) stress, induced by the accumulation of unfolded or malfolded proteins, activates various apoptotic pathways. Crosstalk between the mitochondria and ER plays an essential role in ER stress-mediated cell death. The cytochrome c-dependent apoptotic pathway is activated by ER stress. On the other hand, caspase-12, which is located at the ER, is also activated by excess ER stress and results in cell death in the absence of the cytochrome c-dependent pathway. The predominant apoptotic pathway may differ among cell type and differentiation stage. PMID- 15363496 TI - A review of the development of Respimat Soft Mist Inhaler. AB - Respimat Soft Mist Inhaler (SMI) is a new generation inhaler from Boehringer Ingelheim developed for use with respiratory drugs. The device functions by forcing a metered dose of drug solution through a unique and precisely engineered nozzle (the uniblock), producing two fine jets of liquid that converge at a pre set angle. The collision of these two jets generates the soft mist. The soft mist contains a high fine particle fraction of approximately 65 to 80%. This is higher than aerosol clouds from conventional portable inhaler devices, such as pressurised metered dose inhalers (pMDIs) and dry powder inhalers (DPIs). In addition, the relatively long generation time of the aerosol cloud (approximately 1.5s) facilitates co-ordination of inhalation and actuation--a major problem with pMDIs. These features, together with the slow velocity of the soft mist, result in larger amounts of the drug reaching the lungs and less being deposited in the oropharynx compared with either pMDIs or DPIs. Generation of the soft mist from Respimat SMI is purely mechanical, so propellants are not necessary. The innovative design of Respimat SMI, using water-based drug formulations, ensures patients receive consistent and reliable doses of the drug with each actuation. The device was initially tested in scintigraphic lung deposition studies and produced encouraging results when compared with the chlorofluorocarbon-based pMDI (CFC-MDI). Subsequent clinical studies have confirmed that Respimat SMI is effective and safe in delivering bronchodilators to patients with asthma or chronic obstructive pulmonary disease. PMID- 15363497 TI - New protamine quantification method in microtiter plates using o phthaldialdehyde/N-acetyl-L-cysteine reagent. AB - Protamine is a well-known excipient in pharmaceutics. It represents a peptide consisting of exclusive aliphatic amino acids, hence it cannot be quantified by UV-spectroscopy (lambdamax 280 nm). A new and sensitive quantification method based on the derivatisation of protamine with ortho-phthaldialdehyde (OPA) in the presents of 2-mercaptoethanol (ME) or N-acetyl-L-cysteine (NAC) in basic aqueous solution using 96-well microtiter plates are introduced in this report. The resulting isoindol derivatives reveal a fluorescence excitation (maximum lambdaex 345 nm) and emission (maximum lambdaem 450 nm) spectra. Derivatives of OPA/NAC reagent were found to be useful for protamine quantification in pharmaceutical nanoparticle preparation containing DNA. A sufficient stability of the isoindol derivatives was shown. It was possible to determine protamine free base, protamine sulphate and protamine chloride with limits of detection less than 1.1 microg/ml. PMID- 15363498 TI - Experimental designed optimisation and stability evaluation of dry suspensions with artemisinin derivatives for paediatric use. AB - There is a great need for oral anti-malaria preparations especially for small children, which are easy to administer and keep their stability under tropical conditions. The purpose of this work was therefore to develop a dry suspension, containing one of the artemisinin derivatives, namely artesunate, artemether and dihydroartemisinin using fast wetting suspending agents, i.e. xanthan gum and Avicel CL611. For the optimisation of these two variables, namely the suspending agent's content, a Doehlert design was applied. Via preliminary tests on sedimentation behaviour, the limits of both products were determined, respectively 0.1-0.4% (w/v) and 1.0-2.5% (w/v). As responses, sedimentation as a function of time, viscosity and price of the suspension, were evaluated. The stability tests of the reconstituted suspensions showed bad results for artesunate, even when the pH was adapted. In contrast, dihydroartemisinin showed only 10% degradation within 10 days and artemether was stable at least 21 days. Practically the last one was able to foresee a chemically and physically stable suspension at least during the administration period (5 to 7 days) and was therefore selected for further optimisation concerning taste and appearance. Based on the results of selection tests for the colourant, sweetener and taste masking agent, the following composition was proposed for a suitable dry powder with artemether (AM) as active compound to prepare 100 ml reconstituted suspension: AM 300 mg, Avicel CL611 2 g, xanthan gum 200 mg, crystalline saccharose 35 g, citric acid monohydrate 150 mg, Nipagine 80 mg, Nipasol 20 mg, sodium saccharinate 250 mg, tutti-frutti 250 mg and Sunset yellow 10 mg. PMID- 15363499 TI - Implantable technology for long-term delivery of nalmefene for treatment of alcoholism. AB - Pharmacotherapy treatment for alcoholism is limited by poor compliance, adverse effects, and fluctuating drug levels after bolus administration. A long-term delivery system would improve upon these limitations. The current study describes the characterization of a sustained release implant containing nalmefene, an opioid antagonist, for treatment of alcoholism. Nalmefene was blended with ethylene vinyl acetate (EVA), extruded into 2.8 mm x 27 mm rods, and coated with EVA to optimize release. In vitro release was determined by HPLC, and in vivo release characteristics after subcutaneous implantation into rats were determined by LC-MS/MS analyses. Extrusion produced rods containing 80.09 +/- 6.0 mg nalmefene. In vitro release was high from the uncoated rods, and they were depleted of drug fairly quickly; however EVA coatings maintained release over longer periods. The 25 wt.% coated rods provided in vitro release of 0.36 mg/day/rod, and in vivo release of 0.29 mg/day/rod over 6 months, and showed dose dependent nalmefene plasma concentrations (one rod: 3.33 +/- 0.56 ng/ml, three rods: 10.19 +/- 2.31 ng/ml). After explantation, nalmefene plasma concentrations were undetectable by 6 h. A sustained release nalmefene rod provides 6 months of drug with no adverse effects. PMID- 15363500 TI - In vitro transdermal delivery of the major catechins and caffeine from extract of Camellia sinensis. AB - The aim of this study was to investigate the feasibility of the transdermal delivery of catechins and caffeine from green tea extract. Drug-in-adhesive patches containing 1.35, 1.03, 0.68, and 0.32 mg cm(-2) green tea extract were formulated and the dissolution of (-)-epigallocatechin gallate (EGCg), (-) epigallocatechin (EGC) and (-)-epicatechin (EC) from these was determined. Transdermal delivery was determined across full thickness pig ear skin from saturated solutions of green tea extract in pH 5.5 citrate-phosphate buffer, polyethylene glycol 400 and a 50:50 mixture of the citrate phosphate buffer and polyethylene glycol in addition to patches containing 1.35 mg cm(-2) green tea extract. Dissolution experiments indicated first order release which was dose dependent in respect of the loading level, although the amounts permeated were not always proportional to the amounts in the formulation. The highest percentage permeation of EGCg was found to be from the patch formulation. EGCg, EGC and EC were all successfully delivered transdermally from saturated solutions and adhesive patches containing green tea extract in this study. There was some evidence for the dermal metabolism of EGCg, but after 24 h 0.1% permeated from the patches containing 1.35 mg cm(-2) green tea extract. This was equivalent to the percentage absorbed after intragastric administration of green tea extract in rats. In addition, the concentration of EGCg in the Franz cell receptor chamber after 24 h permeation from the 0.9 cm diameter patch containing 1.35 mg cm(-2) was within the range of Cmax plasma levels achieved after oral dosing of 2.2-4.2 gm(-2) green tea extract. Caffeine was also delivered at concentrations above those previously reported. PMID- 15363501 TI - Kinetics of liposome-encapsulated hemoglobin after 25% hypovolemic exchange transfusion. AB - Liposome-encapsulated hemoglobin (LEH) is being developed as an oxygen therapeutic. In this work, we evaluated a neutral formulation of PEGylated LEH for its circulation and distribution properties in rodent models of 25% hypovolemic exchange transfusion. About 25% of blood in rats and rabbits was exchanged with LEH that had been previously labeled with 99mTc radionuclide. The distribution of 99mTc-LEH was followed by gamma camera imaging and intermittent blood sampling during 48 h, and counting the tissue-associated radioactivity after necropsy at 48 h. On the basis of circulation kinetics, the half-life of 99mTc-LEH in blood was 30 and 39.8 h in rats and rabbits, respectively. Apart from blood, major organs of accumulation of LEH after 48 h included liver (rats, 10.3% and rabbits, 5.4% of injected dose) and spleen (rats, 2.4% and rabbits, 0.8% of injected dose). The results demonstrate that LEH circulates for a prolonged time after administration and that the animals tolerate at least 25% of blood exchange without any distress. Subsequent to the enhanced uptake in the RES, the rats clear LEH from the circulation faster than the rabbits. PMID- 15363502 TI - Precision and detection limit of quality test for amorphous drug in powder X-ray diffractometry. AB - This report puts forward a method of powder X-ray diffractometry to estimate the precision and detection limit of the crystalline component in an amorphous drug. Cefditoren pivoxil (CP) was employed as a model drug. The major error source of the measurement at low crystal contents is shown to be the random noise in a diffraction pattern (halo pattern) of the amorphous material. For the analysis of the noise, the obstructive halo pattern should be eliminated from the observed pattern. The subtraction of the observed halo pattern from another one derived from the same material, extracts the random noise alone, although the noise is amplified by square root 2 times. The noise in the powder X-ray diffractometry was identified as the white noise. On the basis of the stochastic properties of the extracted noise and signal parameters (peak area) of CP, the relative standard deviations (R.S.D.) of the area measurements of the crystalline diffraction peaks were estimated over a wide range of crystal contents without repeated experiments. The detection limit was determined such that the crystal content at detection limit produced 30% R.S.D. of the measurements. The R.S.D. and detection limit obtained from FUMI theory were in good agreement with the results from the repeated measurements. PMID- 15363503 TI - Ultrasonically controlled release and targeted delivery of diclofenac sodium via gelatin magnetic microspheres. AB - In the present work, an attempt was made to target diclofenac sodium to its site of action through magnetic gelatin microspheres. The gelatin magnetic microspheres loaded with 8.9% w/w of diclofenac sodium and 28.7% w/w of magnetite were formulated by emulsification/cross-linking with glutaraldehyde. The formulated microspheres were characterized by particle size distribution, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), X ray diffraction and in vitro release studies. The in vivo distribution and targetability of gelatin magnetic microspheres after i.v. administration were studied in rabbits. The formulated microspheres were below 5 microm and spherical in nature as evidenced by the SEM photographs. DSC and X-ray diffraction studies revealed the absence of drug-polymer interaction. Encapsulated diclofenac sodium was released slowly more than 18 days. Application of sonication, as external stimuli to enhance drug release, during release study, has slightly increased the release rate. The formulated microspheres were injected intravenously after keeping a suitable magnet near the target area. The quantity of drug available at the target and non-target area was determined by HPLC. About 5.5% of injected dose localized near the target organ. Majority of injected dose was recovered from lungs, spleen and liver indicating localization of microspheres in these organs. Further studies are required to improve the targeting efficiency of gelatin microspheres by modifying surface properties to overcome phagocytosis and by selecting suitable particle size to avoid the entrapment of microspheres in non-target organs. PMID- 15363504 TI - Deviation from linearity of drug solubility in ethanol/water mixtures. AB - A new empirical function that describes the deviation from linearity of solubility of a drug in an ethanol/water matrix is applied to the experimental data for 51 compounds. The proposed model is a more accurate predictor of the co solvent solubility profile than a general third order polynomial with the same number of parameters. Both the root mean square error and average absolute error for the proposed model are significantly lower than those of existing models. The model also accurately predicts the fraction of co-solvent that gives maximum solubility (fmax). PMID- 15363505 TI - Effect of plasticization on heparin release from biodegradable matrices. AB - Heparin-loaded polymer films of poly-L-lactide (PLLA) and poly-L-lactide-co glycolide (PLLGA) as well as poly-DL-lactide-co-glycolide (PLGA) were produced. A plasticizer, PEG, was added to the polymers. It was found that the release profile in general consisted of a burst effect, a diffusion-controlled phase and a degradation-controlled phase. The plasticizer accelerated the onset of degradation in all cases, but its effect on the release profile differed significantly depending on the polymer. The plasticizer depressed the burst effect for PLLA, and accelerated the kinetics of the diffusion-controlled phase. For the PLLGA 80/20, however, the plasticizer had no significant effect on the release profile or kinetics. We explain these observations in terms of hydrophilicity and crystallinity effects. PMID- 15363506 TI - Characterization of perivascular poly(lactic-co-glycolic acid) films containing paclitaxel. AB - The objectives of this study were to investigate the use of poly(lactic-co glycolic acid) (PLGA) for the formulation of paclitaxel loaded films and to characterize these films for potential application as perivascular "wraps" to prevent restenosis. Films were manufactured from PLGA blended with either methoxypolyethylene glycol (MePEG) or a diblock copolymer composed of poly(D,L lactic acid)-block-methoxypolyethylene glycol, PDLLA-MePEG (diblock) by solvent evaporation on teflon discs. Elasticity was determined by gravimetric stress/strain analysis. Thermal analysis was determined using differential scanning calorimetry (DSC). Changes in film composition and degradation in aqueous media were determined using gel permeation chromatography (GPC). Paclitaxel release from films was measured by incubation of the films in phosphate buffered saline (PBS) with drug analysis by HPLC methods. The addition of MePEG or diblock to PLGA caused a concentration dependent increase in the elasticity of films, due to plasticizing effects. DSC analysis showed that MePEG and diblock caused a concentration dependent decrease in the glass transition temperature (Tg) of PLGA indicating miscibility of the polymers. When placed in aqueous media, more than 75% of MePEG dissolved out of the PLGA films within 2 days, whereas diblock partitioned slowly and in a controlled manner out of the films. Paclitaxel release from PLGA/MePEG films was very slow with less than 5% of the encapsulated drug being released over 2 weeks. The addition of 30% diblock to paclitaxel loaded PLGA films caused a substantial increase (five- to eight fold) in the release rate of paclitaxel. PLGA films containing 30% diblock and either 1% or 5% paclitaxel were partially or completely degraded following perivascular implantation in rats. PMID- 15363507 TI - ATR-FTIR characterization of transport properties of benzoic acid ion-pairs in silicone membranes. AB - A novel technique based on Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) spectroscopy was used to study the transport of benzoic acid ion-pairs/salts in silicone membranes. The benzoic acid ion-pairs were prepared using various counter-ions with different degrees of lipophilicity, e.g. triethylamine (TA), diethylamine (DE), tert-butylamine (t-BA), 2-amino-2-methyl propanol (AMP), and 2-amino-2-methyl-propanediol (AMPD). Silicone membrane, treated or untreated with propylene glycol (PG), was placed on the surface of a ZnSe crystal and the transport solution was applied to the upper surface of the membrane. A mathematical model, based on Fick's second law describing the build up of permeant concentration at the membrane/crystal interface with time was applied to determine diffusion coefficients. Absorption due to the acid (1700 cm( 1)) or benzoate anion (1555 cm(-1)) was observed at different regions without the interference from PG or silicone membrane. Benzoate anion, a charged species, was observed to permeate the membrane. The permeation of benzoate anion from sodium benzoate and polar ion-pairs of AMP and AMPD was very low in contrast to their high-saturated concentrations in PG as compared to the t-BA ion-pair. This indicated that benzoate anion preferentially permeates the membrane as an ion pair rather than a single anion; otherwise its permeation should correspond to its concentration in PG instead of the lipophilicity of the ion-pairs. Additionally, the diffusion coefficient values of benzoic acid and benzoate anions through the treated and untreated membranes were not statistically different. PMID- 15363508 TI - Grouping solvents by statistical analysis of solvent property parameters: implication to polymorph screening. AB - The success rate of discovering new polymorphs by crystallization from solution may be increased if solvents with diverse properties are used during initial polymorph screening. In this study, eight solvent parameters, including hydrogen bond acceptor propensity, hydrogen bond donor propensity, polarity/dipolarity, dipole moment, dielectric constant, viscosity, surface tension and cohesive energy density (equal to square of solubility parameter), of 96 solvents were collected. Using the cluster statistical analysis of the parameters, these 96 solvents were separated into 15 solvent groups. Such solvent groups may provide guidelines for the judicious choice of solvents with diverse properties for polymorph screening. PMID- 15363509 TI - (S)-Nitroxycarnitine nitrate production from (S)-carnitine by using acetic anhydride/nitric acid/acetic acid mixtures: safety assessment. AB - The present work aims at assessing both thermodynamic and kinetic parameters of the esterification process of the (S)-carnitine, using calorimetric techniques. The use of the system acetic anhydride/nitric acid/acetic acid as esterifying agent and the explosive behaviour of nitric esters lead to safety considerations that have been investigated by hypothesizing some common process deviations. In particular, it has been investigated in adiabatic conditions both the batch addition of acetic anhydride and the effect of an initial temperature higher than those required by the process. PMID- 15363510 TI - Synthesis, characterization and thermal studies on furazan- and tetrazine-based high energy materials. AB - This paper reports the synthesis of high energy materials (HEMs) viz. 3,3' diamino-4,4'-azoxyfurazan (DAAF), 3,3s'-azobis(6-amino-1,2,4,5-tetrazine) [DAAT] and 1,4-dihydrazino tetrazine (DHTz). The products obtained were characterized by IR, 1H NMR, 13C NMR and mass spectra. Thermolysis of these compounds carried out by applying TG-DTA and DSC techniques indicated that the thermal stability of DAAF and DAAT was in the temperature range of 230-250 degrees C, whereas that of DHTz was up to approximately 140 degrees C. TG-FTIR of gaseous products of these compounds suggests the evolution of NH2CN/NH3 and HCN as major decomposition products. The impact and friction sensitivity data revealed that DAAF is insensitive to mechanical stimuli whereas DAAT and DHTz are vulnerable to impact stimuli. The cyclic voltammetric studies brought out that, DAAF, DAAT and DHTz are electroactive compounds and thereby can be detected at even low concentration at pH 7 and 13. The theoretical predictions of explosive power of DAAF, DAAT and DHTz alone and their combinations with well-known insensitive high explosives using Becker-Kistiakowsky Wilson (BKW) code as well as that of propellants based on them by NASA-CEC-71 suggest their potential in specific systems. PMID- 15363511 TI - Synthesis, characterization, thermolysis and performance evaluation of mercuric-5 nitrotetrazole (MNT). AB - Mercuric-5-nitrotetrazole (MNT) was synthesized on using a reported method. The product having bulk density of 1.5 g/cm3, was obtained during this work using mercuric nitrate doped with additives such as cephol/dextrin in the process. Synthesized MNT was characterized by metal content analysis, IR and ESCA. The DTA profile indicated the thermal stability of MNT up to 200 degrees C. It revealed its higher thermally sensitive [thermal sensitive figure (S) approximately 0.8] in comparison to that of service lead azide (SLA) [S approximately 0.4]. Percussion sensitivity data also showed higher sensitivity of MNT. However, it was found less friction sensitive than SLA. The chemical stability of MNT in a carbon dioxide environment was evaluated in comparison to SLA by determining mercury (gravimetrically) and lead azide (volumetrically) contents respectively. Results obtained indicated that no discernable changes occurred in MNT, even after storage for 90 days while in case of SLA, drastic change in lead azide content was observed. IR spectra of MNT sample stored in a closed aluminum dish for 5-10 years could be superimposed on that of the freshly prepared MNT sample. The performance of MNT filled detonator no. 27 assessed in terms of extent of damage on a witness plate was found equivalent to that of the standard ASA (azide, styphynate and aluminium) composition filled detonator. PMID- 15363512 TI - 1,3,3-Trinitroazetidine (TNAZ), a melt-cast explosive: synthesis, characterization and thermal behaviour. AB - 1,3,3-Trinitroazetidine (TNAZ) has been prepared at a laboratory scale in HEMRL. The structure of the compound has been confirmed by IR, NMR, mass, elemental analysis and by X-ray crystallography. HPLC technique has been employed to confirm the purity of TNAZ (>99%). The compound is further characterized by thermal techniques and is found to undergo limited decomposition at its melting point. Small scale sensitivity tests have also been carried out and the results show that TNAZ is significantly more sensitive to mechanical stimuli than TNT. PMID- 15363513 TI - Parametric analyses of diffusion of activated sources in disposal forms. AB - The primary objective of a low-level radioactive waste disposal facility is to isolate low-level radioactive waste from the public and the environment until the radionuclides in the waste have decayed to levels at which the hazard is negligible. Two fundamental concerns must be addressed when attempting to isolate low-level waste in a disposal facility on land. The first concern is isolating the waste from water, or hydrologic isolation. The second is preventing movement of the radionuclides out of the disposal facility, or radionuclide migration. Particularly, we have investigated here the latter scenario. Empirical curves describing the progression of the altered zone are obtained and they are used to define the correspondent altered thickness in the numerical calculations for a cement sample. Subsequent numerical analyses of diffusion of activated sources in waste disposals are considered for cemented containers. The adopted DAMAGE code takes into account a mass conservation equation and the linear momentum balance equation for the multiphase material. The mathematical model is based on the theory developed by Bazant for concretes and geomaterials; the fluid phases are considered as a unique mixture interacting with a solid phase. Short- and long term diffusion analyses are performed for different characteristics of the grout and the results are presented in terms of radionuclides concentration. Indications on the minimum grout thicknesses able to resist to radioactive fluxes up to 1000 years are given. PMID- 15363514 TI - Studies on lead-free initiators: synthesis, characterization and performance evaluation of transition metal complexes of carbohydrazide. AB - Cobalt, nickel and zinc tris(carbohydrazide) perchlorates (CoCP, NiCP and ZnCP) as well as copper bis(carbohydrazide) perchlorate (CuCP) of desired morphology and bulk density (0.85-0.95 g/cm3) have been synthesized during this work. The synthesis was carried out by addition of the aqueous solution of cobalt/nickel/copper/zinc perchlorates to the aqueous solution of carbohydrazide. The products were characterized by the metal content analysis and FTIR. The IR spectra and ESCA brought out the coordination of both the end amino groups of carbohydrazide with the central metal ion. Differential thermal analysis (DTA) curve indicated that CoCP, NiCP and ZnCP are thermally stable in the temperature range of 220-285 degrees C, unlike CuCP (120 degrees C). The activation energy determined by TG measurements was found to be 140-180 kJ/mol for CoCP, NiCP and ZnCP. Sensitivity data revealed their sensitivity to friction stimuli (1 kg). Impact sensitivity test results corresponded to h50% of 50-60 cm with the exception of CuCP (h50%, 11 cm). In order to assess the performance as detonants, the selected compounds were detonated on a lead witness plate of 3 mm thickness using fuse wire as well as evaluated in conjunction with tetryl in detonator No. 27 tube. The results obtained in terms of extent of damage to witness plate were on par with the standard detonator No. 27 containing azide, styphynate and aluminium metal (ASA) composition. PMID- 15363515 TI - Thermochemical properties and non-isothermal decomposition reaction kinetics of 3,4-dinitrofurazanfuroxan (DNTF). AB - The constant-volume combustion energy, DeltacU (DNTF, s, 298.15K) and kinetic behavior of the exothermic decomposition reaction of the title compound (DNTF) are determined by a precise rotating bomb calorimeter and DSC, respectively. Its standard enthalpy of combustion, DeltacHmtheta (DNTF, s, 298.15K), standard enthalpy of formation, DeltacHmtheta (DNTF, s, 298.15K) and kinetic parameters of the major exothermic decomposition reaction in a temperature-programmed mode [the apparent activation energy (Ea) and pre-exponential factor (A)] are calculated. The values of DeltacU (DNTF, s, 298.15K), DeltacHmtheta (DNTF, s, 298.15K), and DeltacHmtheta (DNTF, s, 298.15K) of DNTF are -9733.96 +/- 8.59 Jg(-1), -3018.29 +/- 2.68 kJ mol(-1), and 657.23 +/- 2.70 kJ mol(-1), respectively. The kinetic model function in integral form and the value of E(a) and A of the major exothermic decomposition reaction of DNTF are 1-(1-alpha)1/3, 177.03 kJ mol(-1) and 10(13.68)s(-1), respectively. The critical temperature of thermal explosion of DNTF is 240.6 degrees C. PMID- 15363516 TI - Removal and recovery of mercury(II) from hazardous wastes using 1-(2 thiazolylazo)-2-naphthol functionalized activated carbon as solid phase extractant. AB - As a part of removal of toxic heavy metals from hazardous wastes, solid phase extraction (SPE) of mercury(II) at trace and ultra trace levels was studied using 1-(2-thiazolylazo)-2-naphthol (TAN) functionalized activated carbon (AC). The SPE material removes traces of mercury(II) quantitatively in the pH range 6.0 +/- 0.2. Other parameters that influence quantitative recovery of mercury(II), viz. percent concentration of TAN in AC, amount of TAN-AC, preconcentration time and volume of aqueous phase were varied and optimized. The possible means of removal of Hg(II) from other metal ions that are likely to be present in the wastes of the chloroalkali industry is discussed. The potential of TAN-functionalized AC SPE material for decontaminating mercury from the brine sludge and cell house effluent of a chloralkali plant has been evaluated. PMID- 15363517 TI - Dye removal from wastewater using activated carbon developed from sawdust: adsorption equilibrium and kinetics. AB - Mahogany sawdust was used to develop an effective carbon adsorbent. This adsorbent was employed for the removal of direct dyes from spent textile dyeing wastewater. The experimental data were analysed by the Langmuir and Freundlich models of adsorption. Equilibrium data fitted well with the Langmuir model. The rates of adsorption were found to conform to the pseudo-second-order kinetics with good correlation. The equilibrium adsorption capacity of the sawdust carbon was determined with the Langmuir equation as well as the pseudo-second-order rate equation and found to be >300 mg dye per gram of the adsorbent. The most ideal pH for adsorption of direct dyes onto sawdust carbon was found to be 3 and below. The results indicate that the Mahogany sawdust carbon could be employed as a low cost alternative to commercial activated carbon in the removal of dyes from wastewater. PMID- 15363518 TI - Photocatalytic oxidation of VX simulant 2-(butylamino)ethanethiol. AB - Photocatalytic oxidation of 2-(butylamino)ethanethiol (BAET) was undertaken in aqueous suspension of TiO2 Hombikat UV 100 and Degussa P25 under different initial reaction conditions in order to determine the best parameters for the fastest mineralization of the substrate. BAET is considered to be a simulant for the VX chemical warfare agent. The application of ultrasound had only a small positive effect on the BAET photocatalytic degradation. The highest mineralization rate of 0.433 mg/(l min) was found in unbuffered TiO2 Degussa P25 suspension with initial pH value of about 9.4, TiO2 concentration 500 mg/l and the initial BAET concentration 1000 mg/l. Decreasing of the initial solution pH to 6.1 or below stops the mineralization of BAET while increasing of pH to about 11 drastically changed the degradation profile. At this initial pH, the first 100 min of reaction led to only oxidation of sulfur moiety and organic intermediates accumulated in the solution. Thereafter, mineralization of the products started. The main detected volatile product was butyl aldehyde and the main polar one was 2-(butylamino) acetic acid. In the case of TiO2 Hombikat UV 100, conversion of TOC at initial pH 11 exceeded that at initial pH 9.1. For Degussa P25, the starting pH 9.4 was the best for TOC conversion. The results can be used for treatment of water from pollutants with aliphatic nitrogen and sulfur atoms. PMID- 15363519 TI - Adsorption of Pb(II) from aqueous solution by Azadirachta indica (Neem) leaf powder. AB - An adsorbent was developed from the mature leaves of the Neem (Azadirachta indica) tree for removing Pb(II) from water. Adsorption was carried out in a batch process with several different concentrations of Pb(II) by varying amount of adsorbent, pH, agitation time and temperature. The uptake of the metal was very fast initially, but gradually slowed down indicating penetration into the interior of the adsorbent particles. Both first-order and second-order kinetics were tested and it was found that the latter gave a better explanation. The experimental data closely followed both Langmuir and Freundlich isotherms. The adsorbent had a considerably high Langmuir monolayer capacity of 300 mg/g. A small amount of the adsorbent (1.2 g/L) could remove as much as 93% of Pb(II) in 300 min from a solution of concentration 100mg/L at 300 K. The adsorption continuously increased in the pH range of 2.0-7.0, beyond which the adsorption could not be carried out due to the precipitation of the metal. The adsorption was exothermic at ambient temperature and the computation of the parameters, DeltaH, DeltaS and DeltaG, indicated the interactions to be thermodynamically favourable. PMID- 15363520 TI - Effects of nickel(II) addition on the activity of activated sludge microorganisms and activated sludge process. AB - The effects of Ni(II) in a synthetic wastewater on the activity of activated sludge microorganisms and sequencing batch reactor (SBR) treatment process were investigated. Two parallel lab-scale SBR systems were operated. One was used as a control unit, while the other received Ni(II) concentrations equal to 5 and 10 mg/l. The SBR systems were operated with FILL, REACT, SETTLE, DRAW and IDLE modes in the time ratio of 0.5:3.5:1.0:0.75:0.25 for a cycle time of 6 h. The addition of Ni(II) into SBR system caused drastically dropped in TOC removal rate (k) and specific oxygen uptake rate (SOUR) by activated sludge microorganisms due to the inhibitory effects of Ni(II) on the bioactivity of microorganisms. The addition of 5 mg/l Ni(II) caused a slight reduction in TOC removal efficiency, whereas 10 mg/l Ni(II) addition significantly affected the SBR performance in terms of suspended solids and TOC removal efficiency. Termination of Ni(II) addition led to almost full recovery of the bioactivity in microorganisms as shown in the increase of specific oxygen uptake rate (SOUR) and SBR treatment performance. PMID- 15363521 TI - Electrochemical treatment of industrial wastewater. AB - This paper presents the results of the treatment of phenolic compounds containing wastewater generated from phenol-formaldehyde resin manufacturing, oil refinery and bulk drug manufacturing industries by electrochemical method. Experiments were conducted at a fixed current density of 5.4 A/dm2 using Ti/TiO2-RuO2-IrO2 electrode and an undivided reactor. During the various stages of electrolysis, parameters such as COD and TOC concentrations were determined in order to know the feasibility of electrochemical treatment. Adsorbable organic halogens (AOX) were detected at high concentrations during the electrolytic treatment of the effluents. However, it was observed that increasing the electrolysis time bring down the AOX concentration to lower levels. Energy consumption and current efficiency during the electrolysis were calculated and presented. The present study proves the effectiveness of electrochemical treatment for highly concentrated bio-refractory organic pollutants present in the industrial wastewater. PMID- 15363522 TI - Influence of the microporosity and surface chemistry of polymeric resins on adsorptive properties toward phenol. AB - In this work, the effects of the microporosity and chemical surface of polymeric adsorbents on adsorptive properties of phenol were investigated. Textural parameters of four kinds of polymeric resins, namely AB-8, D4006, NKA-II and D16 resin, were separately measured by ASAP 2010. The surface chemistry of these polymeric resins was determined by means of inverse gas chromatography (IGC) and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS). Static equilibrium adsorption experiments were carried out to obtain the isotherms of phenol on the polymeric resins. It was shown that NKA-II and AB-8 resin possessed relatively high BET surface areas and micropore volumes, while D4006 and D16 resin possessed comparatively low BET surface areas and micropore volumes. The results of IGC experiments revealed that NKA-II resin had extraordinary high specific component of the free energy of adsorption both for polar acetone and benzene probe, and thus extraordinary strong surface polarity compared to the other polymeric resins. It was also found that the isotherm of phenol on NKA-II was much higher than that on the other polymeric resins due to its strongest surface polarity and largest micropore volume among four kinds of resins. These experimental observations indicated that adsorption of phenol on the polymeric resins depended greatly on their microporosity and surface chemistry. The well developed microporosity and the strong surface polarity would improve the adsorption of phenol on the polymeric resins. PMID- 15363523 TI - Biodegradation of 2,4-dinitrotoluene using poultry litter leachate. AB - Various micro-organisms are known to degrade 2,4-dinitrotoluene (DNT) through pathways involving intermediates. Addition of nutrients stimulates the biodegradation process. Poultry litter has a consortia of micro-organisms along with many nutrients. The objective was to study the DNT biodegradation using poultry litter in an aqueous medium. Complete biodegradation of 10-50 ppm 2,4-DNT solutions was observed after two days interaction with poultry litter leachate without the formation of any intermediates. No degradation was observed using autoclaved litter leachate. PMID- 15363524 TI - Heavy metal stabilization in municipal solid waste incineration flyash using heavy metal chelating agents. AB - Heavy metal chemical stabilization with synthesized heavy metal chelating agent was assessed for flyash from municipal solid waste incinerator. Flyash can contain heavy metals (e.g. Pb, Cd) which can leach. A new kind of heavy metal chelating agent showed more attractive competition than inorganic chemicals in stabilizing flyash. The synthesizing method of this kind of heavy metal chelating agent was explained in this paper, and the technology process and treatment efficiency of the chelating agent in treating flyash were experimentally studied, which was compared with the results of inorganic chemical agents such as sodium sulfide and lime. The heavy metals in flyash were stabilized more effectively by using heavy metal chelating agents than by using sodium sulfide and lime, furthermore, the stabilized products using the chelating agents can meet the landfill disposal controlling standards for heavy metal waste. pH-dependent leaching experiment showed the stabilized flyash by treatment with heavy metal chelating agent could keep long-term stabilization within a broad range of pH value. Thus, the risk of secondary pollution for the stabilized products was reduced dramatically when the environment condition changes during its disposal period. PMID- 15363525 TI - Biological pre-treatment of wastewater containing sulfate using anaerobic immobilized cells. AB - Biological reduction of sulfate to sulfide using sulfate reducing bacteria (SRB) was investigated. A respirometer was used to study the sulfide toxicity in the systems fed glucose, the results showed that sulfide would start to inhibit methanogens when the dissolved sulfide and total sulfide concentrations were 276.4 and 304.6 mg/L, respectively. When chemostats were used to study the Monod kinetic coefficients, Y, kd, Ks, and k were 0.36 mg VSS (volatile suspended solids) using SRB/mg SO4-S, 0.05/day, 147.30 mg SO4-S/L, and 6.50 mg SO4-S/mg VSS using SRB-d, respectively. Using pure cultural techniques, SRB were found to be 29.45% of the VSS in the chemostats. Sulfate removal using an upflow anaerobic filter packed with immobilized cells was also investigated. Under sulfate loading rates of 0.2 and 0.4 g SO4-S/L day, and a hydraulic retention time (HRT) of 2 days, a sulfate removal efficiency greater than 93% could be achieved. When the filter was operated under COD (chemical oxygen demand)/S from 10/1 to 5/1 and HRTs of 2, 1 and 0.5 days, sulfate removal efficiency was between 98.1 and 70.9%. It is believed that protection by the immobilized cell structure caused the microbial cells in the filter to tolerate higher dissolved sulfide (447.8 mg/L) and total sulfide (940.3 mg/L) levels, allowing a much higher biomass concentration (13.2-13.5 g VSS/L) to be reached. PMID- 15363526 TI - Stabilization of high mercury contaminated brine purification sludge. AB - The highly leachable mercury contaminants of brine purification sludge (BPS) generated from the Hg-cell electrolysis process in chlorine production can be stabilized in the treatment procedure employing ferric-lignin derivatives (FLD) (Ligmet binder) and Portland cement (PC). The stabilization effectiveness has been examined by time-based multiple toxicity characteristic leaching procedure (TCLP) tests and sequential TCLP tests. In a period of 50 days, the multiple TCLP tests showed a variation of less than 90 microg l(-1) for the leachable mercury level, and the sequential TCLP tests for the same sample displayed a declining TCLP mercury level. Based on this study, the stabilization of approximately 2000 t of brine purification sludge has been successfully processed with the ferric lignin derivatives treatment. PMID- 15363527 TI - Evaluation on thermal explosion induced by slightly exothermic interface reaction. AB - An asphalt-salt mixture (ASM), which once caused a fire and explosion in a reprocessing plant, was prepared by imitating the real bituminization process of waste on a lab scale to evaluate its actual thermal hazards. Heat flux reaction calorimeters were used to measure the release of heat for the simulated ASM at a constant heating rate and at a constant temperature, respectively. Experimental results show that the reaction in the ASM below about 250 degrees C is a slightly exothermic interface reaction between the asphalt and the salt particles contained in the asphalt, and that the heat release rate increases sharply above about 250 degrees C due to melting of the salt particles. The reaction rates were formulated on the basis of an assumed reaction model, and the kinetic parameters were determined. Using the model with the kinetic parameters, temperature changes with time and drum-radius axes for the ASM-filled drum were numerically simulated assuming a one-dimensional infinite cylinder system, where the drum was being cooled at an ambient temperature of 50 degrees C. The minimum filling temperature, at which the runaway reaction (MFTRR) can occur for the simulated ASM in the drum is about 194 degrees C. Furthermore, a very good linear correlation exists between this MFTRR and the initial radius of salt particles formed in the bituminization product. The critical filling temperature to the runaway reaction is about 162 degrees C for the asphalt-salt mixture, containing zero-size salt particles, filled in the same drum at an ambient temperature of 50 degrees C. Thus, the runaway reaction will never occur in the drum filled with the asphalt-salt mixture under the conditions of the filling temperature below 162 degrees C and a constant ambient temperature of 50 degrees C. As a consequence, the ASM explosion occurred in the reprocessing plant likely was due to a slightly exothermically reaction and self heating. PMID- 15363528 TI - Removal of trichloroethylene in reduced soil columns. AB - A continuous soil column experiment was conducted to investigate reductive dechlorination of trichloroethylene (TCE) in soil system reductively manipulated by three types of reductants (Fe(II), dithionite, and Fe(II) + dithionite (combined treatment of Fe(II) and dithionite)). The soil column reduced by Fe(II) + dithionite has the greatest bed volumes (51.8) treated to breakthrough indicating that the combined treatment of Fe(II) and dithionite is more effective for the reductive dechlorination of TCE in the reduced soil column than the separate treatment of Fe(II) or dithionite. The measured bed volumes to breakthrough in control and treated soil columns were similar to the estimated bed volumes based on the result of batch kinetic experiment, differing by a factor of 0.96-1.02. The relative concentration of bromide (non-reactive tracer) reached the approximate value of 1 between 0.87 and 1.03 bed volume. C2 hydrocarbons (acetylene, ethylene, and ethane) were observed as transformation products in the effluents of soil columns treated by the reductants. However, no chlorinated intermediates were observed at the concentrations above detection limits throughout the experiment. Chloride was observed in the effluents of soil columns reduced by dithionite and Fe(II) + dithionite. PMID- 15363529 TI - Microbiological degradation of a spent offset-printing developer. AB - To decontaminate spent offset-printing developer Polychrome 4003, several microorganisms were separated from the soil that has been used for developer dumping for 3 years. Two organism cultures were isolated and identified to genus Geomyces pannorum and Bacteria spp. These organisms, as well as commercial Septic Gobbler (SG) bacteria, were used to decontaminate the developer. Reduction of both the chemical oxygen demand (COD) and the amount of total identified organic compounds reached 30% after 40 day treatment of waste suspension by G. pannorum and Bacteria spp. A substantially higher degree of COD reduction by approximately 80% and the total amount of identified organic compounds by approximately 90% was achieved when SG bacteria have been applied for the same period. According to a rapid electrophysiological test with macrophytic algae Nitellopsis obtusa, the toxicity of spent offset-printing developer Polychrome 4003 was classified as extremely toxic (>100 toxic units, T.U.), and it remained at the same level after treatment with G. pannorum and Bacteria spp. More effective biodegradation with SG bacteria diminished toxicity substantially. PMID- 15363530 TI - The removal of phenolic compounds from aqueous solutions by organophilic bentonite. AB - The adsorption of p-chlorophenol (p-CP) and p-nitrophenol (p-NP) on organophilic bentonite (dodecylammonium bentonite, DDAB) was studied as a function of solution concentration and temperature. The observed adsorption rates were found to be equal to the first-order kinetics. The rate constants were calculated for temperatures ranging between 25.0-35.0 degrees C at constant concentration. The adsorption energies, E and adsorption capacity, (qm), for phenolic compounds adsorbed to organophilic bentonite were estimated by using the Dubinin Radushkevic equation. Thermodynamic parameters from the adsorption isotherms of p CP and p-NP on organophilic bentonite were determined. These isotherms were modeled according to Freundlich and Dubinin-Radushkevic adsorption isotherms and followed the V-shaped isotherm category with two steps. The amount of adsorption was found to be dependent on the relative energies of adsorbent-adsorbate, adsorbate-solvent and adsorbate-adsorbate interaction. PMID- 15363531 TI - Adsorption of acid dye from water onto pristine and acid-activated clays in fixed beds. AB - The adsorption of an acid dye from water onto pristine and HCl-activated montmorillonites in fixed beds was investigated. Experiments were carried out as a function of liquid flow rate, initial dye concentration, and bed height. The adsorption capacity of acid dye onto pristine clay could be largely improved when the clay was activated by HCl. A mass transfer model that involves only two parameters, tau (50% breakthrough time) and kappa (adsorption rate constant), was proposed. This model could satisfactorily describe the measured breakthrough curves of acid dye in fixed beds (standard deviation <6%). It was shown that the value of tau decreased with increasing liquid flow rate. The effect of the type of clay (pristine, acid-activated) on the values of kappa, tau, and adsorption capacity was discussed, and the application potential of acid-activated clay for adsorption removal of acid dye from water was also demonstrated. PMID- 15363532 TI - Decolorization of azo dye acid black 1 by the UV/H2O2 process and optimization of operating parameters. AB - An advanced oxidation process, UV/H2O2, was applied for decolorization of a di azo dye (acid black 1). The effects of operating parameters such as hydrogen peroxide dosage, UV dosage and initial dye concentration, on decolorization have been evaluated. The acid black 1 solution was completely decolorized under optimal hydrogen peroxide dosage of 21.24 mmol/l and UV dosage of 1400 W/l in less than 1.2 min. The decolorization rate followed pseudo-first order kinetics with respect to the dye concentration. The rate increased linearly with volumetric UV dosage and nonlinearly with increasing initial hydrogen peroxide concentration. It has been found that the degradation rate increased until an optimum of hydrogen peroxide dosage, beyond which the reagent exerted an inhibitory effect. For real case application, an operation parameter plot of rate constant was developed. To evaluate the electric power and hydrogen peroxide consumption by UV/H2O2 reactor, 90% color removal was set as criteria to find the balance between both factors. PMID- 15363533 TI - The leaching behavior of incinerator bottom ash as affected by accelerated ageing. AB - Different artificial ageing treatments were applied to fresh incinerator bottom ash with the aim of promoting/accelerating the natural reactions occurring over time on the mineral phases in the material. The weathering treatments included accelerated carbonation, treatment with air and treatment with nitrogen gas. Both fresh and treated bottom ashes were characterized for their mineralogical composition, acid neutralization capacity and leaching behavior of metals. The results were compared to those obtained from similar characterization of bottom ash samples weathered under atmospheric conditions for 4 years. The findings from the experimental campaign indicated that both the release of metals and the type of solubility-controlling solid phases varied as a function of the duration and type of ageing treatment. The most appreciable influence on mineralogy, acid neutralization behavior and metal leaching was observed for the 48 h carbonation treatment. It was also observed that 4-day aeration of bottom ash was capable of producing similar buffering behavior to that displayed by naturally aged bottom ash. PMID- 15363534 TI - Cr(VI) reduction in aqueous solutions by siderite. AB - Hexavalent chromium is a common and toxic pollutant in soils and wastewaters. Reduction of the mobile Cr(VI) to less mobile and less toxic Cr(III) is a solution for decontamination of industrial effluents. In this study, the reduction of hexavalent chromium in aqueous solutions by siderite was investigated. The influences of amount of acid, contact time, siderite dosage, initial Cr(VI) concentration, temperature and particle size of siderite have been tested in batch runs. The process was found to be acid, temperature and concentration dependent. The amount of acid is the most effective parameter affecting the Cr(VI) reduction since carbonaceous gangue minerals consume acid by side reactions. The highest Cr(VI) reduction efficiency (100%) occurred in the 50 mg/l Cr(VI) solution containing two times acid with respect to stoichiometric amount of Cr(VI) and at the conditions of siderite dosage 20 g/l, contact time 120 min and temperature 25 degrees C. Reduction efficiency increased with increase in temperature and decrease in particle size. The reduction capacity of siderite was found to be 17 mg-Cr(VI)/g. PMID- 15363535 TI - Removal of nickel(II) ions from aqueous solution using crab shell particles in a packed bed up-flow column. AB - This paper investigates the ability of crab shell to remove nickel(II) ions from aqueous solution in a packed bed up-flow column with an internal diameter of 2 cm. The experiments were performed with different bed heights (15-25 cm) and using different flow rates (5-20 ml/min) in order to obtain experimental breakthrough curves. The bed depth service time (BDST) model was used to analyze the experimental data and the model parameters were evaluated. The column regeneration studies were carried out for seven sorption-desorption cycles. The elutant used for the regeneration of the sorbent was 0.01 M EDTA (disodium) solution at pH 9.8 adjusted using NH4OH. Due to continuous usage of crab shell, a performance loss was observed as the breakthrough curves become more flattened also indicated by the broadened mass transfer zone. The breakthrough time decreased uniformly from 28.1 to 9.5 h as the cycles progressed from one to seven, whereas nickel uptake remained approximately constant throughout the seven cycles. The life-factors for crab shell in terms of critical bed length and breakthrough time were found to be 1.1 cm/cycle and 0.17 per cycle, respectively. The elution efficiency was greater than 99.1% in all the seven cycles. The pH profiles during both sorption and desorption process were also reported. In sorption cycles, there was a sudden raise in pH in the early part of the process and then the pH decreased as the time progressed. In desorption cycles, pH decreased in initial stages and followed by gradual increase in pH, which eventually reached the pH of the inlet elutant. PMID- 15363536 TI - Recovery of cyanide in gold leach waste solution by volatilization and absorption. AB - In this study, the effects of pH, time and temperature in regeneration of cyanide in the leaching waste solution of gold production from disseminated gold ore by cyanidation process were investigated and the optimum conditions, consumptions and cyanide recovery values were determined. The sample of waste solution containing 156 mg/l free CN- and 358 mg/l total CN-, that was obtained from Gumushane-Mastra/Turkey disseminated gold ores by cyanidation and carbon-in-pulp (CIP) process under laboratory conditions was used in the experiments. Acidification with H2SO4, volatilization of hydrogen cyanide (HCN) with air stripping and absorption of HCN in a basic solution stages were applied and under optimum conditions, 100% of free cyanide and 48% of complex cyanide and consequently 70% of the total cyanide in the liquid phase of gold leach effluent are recovered. PMID- 15363537 TI - Application of solid-phase extraction to determination of polycyclic aromatic hydrocarbons in sewage sludge extracts. AB - The study presents the efficiency of sewage sludge sample clean-up with the application of SPE columns with various types of adsorbents. Six columns were tested: C8-octyl, C18 PolarPlus, C18-octadecyl, silicagel (SG), phenyl, cyano. The highest efficiency of recovery was observed for C18-octadecyl. Then, using C18, the method was optimised by changing the following parameters: eluent type and volume, column drying and effect of washing of cartridge. PMID- 15363538 TI - Two possible pathways for the release of arsenic during pyrolysis of chromated copper arsenate (CCA)-treated wood. AB - The behavior of arsenic during pyrolysis at temperatures between 135 and 500 degrees C has been widely studied. We examined the release of arsenic during the pyrolysis of chromated copper arsenate (CCA)-treated wood, and here we propose two models for the volatilization of arsenic: (a) a reacted arsenic compound, chromium arsenate, in CCA-treated wood was initially decomposed to arsenic pentoxide, which then appeared to be released at around 400-500 degrees C as a form of arsenic trioxide; and (b) an unreacted arsenic compound, arsenic pentoxide, in CCA-treated wood was released at much lower temperatures than reacted arsenic compound, as a form of arsenic trioxide. In order to restrain the release of arsenic during pyrolysis, it is possible to pyrolyze CCA-treated wood carefully at temperatures around 300-350 degrees C only if the wood contains little unreacted arsenic compound. Under these conditions, almost all toxic elements remain in the pyrolysis residue, and a weight reduction of the wood of approximately 50% can be obtained. On the other hand, the release of arsenic can be achieved easily using an unreacted arsenic compound under the same pyrolysis conditions. However, in the present study it was difficult to determine the accurate release point of arsenic in both cases due to the level of experimental uncertainty. PMID- 15363539 TI - Genetic progression of metastatic melanoma. AB - Melanoma progression is well defined in its clinical, histopathological and biological aspects, but the molecular mechanism involved and the genetic markers associated to metastatic dissemination are only beginning to be defined. The recent development of high-throughput technologies aimed at global molecular profiling of cancer is switching on the spotlight at previously unknown candidate genes involved in melanoma, such as WNT5A and BRAF. In fact, several tumor suppressors and oncogenes have been shown to be involved in melanoma pathogenesis, including CDKN2A, PTEN, TP53, RAS and MYC, though they have not been related to melanoma subtypes or validated as prognostic markers. Here, we have reviewed the published data relative to the major genes involved in melanoma pathogenesis, which may represent important markers for the identification of genetic profiles of melanoma subtypes. PMID- 15363540 TI - Cancer chemopreventive activity of 3beta-methoxyserrat-14-en-21beta-ol and several serratane analogs on two-stage mouse skin carcinogenesis. AB - 3beta-Methoxyserrat-14-en-21beta-ol (1) and 3alpha-methoxyserrat-14-en-21beta-ol (2) are the most abundant triterpenoids from two Picea plants, Picea jezoensis (Sieb. et Zucc.) Carr. var. jezoensis and P. jezoensis (Sieb. et Zucc.) Carr. hondoensis (Mayr) Rehder, and the total yield of 1 and 2 reach over 1/3 of the chloroform extract of the above two plants. This study deals with the potential of anti-tumor promoting activity of 1 and results of the assay of 22 synthetic serratane-type triterpenoids (6)-(27) derived from 1, 2, 21-episerratenediol (3), diepiserratenediol (4) and 13alpha,14alpha-epoxy-3beta-methoxyserratan-21beta-ol (5) to discuss the structure-activity relationship. As a preliminary evaluation of their potential to inhibit tumor promotion, the inhibitory effects on Epstein Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) were used. All compounds except for 12 and 19 showed potent inhibitory effects on EBV-EA induction (100% inhibition at 1000 mol ratio/TPA), their effects being stronger than that of a positive control oleanolic acid. Compounds 1, 13, 14, 18, 20 and 26 were selected to examine the effect on the in vivo two-stage mouse skin carcinogenesis test induced by 7,12 dimethylbenz[a]anthracene (DMBA) as an initiator and TPA as a promoter. The most abundant triterpenoid 1 and the synthetic compounds 13 and 14 were found to exhibit the excellent anti-tumor promoting activity in the in vivo carcinogenesis test, and compounds 18, 20 and 26 also showed strong inhibitory effects. PMID- 15363541 TI - Relation of epidermal growth factor receptor, phosphorylated-Akt, and hypoxia inducible factor-1alpha in non-small cell lung cancers. AB - The role of the epidermal growth factor receptor (EGFR)/Posphorylated-Akt (P-Akt) signaling axis in regulating hypoxia-inducible factor-1alpha (HIF-1alpha) expression in vivo is not well understood and is of potential clinical importance since the extent of hypoxia in the tumor environment is thought to be an important determination of resistance to chemotherapy and radiotherapy. We performed the immunohistochemical studies in 80 patients with non-small cell lung cancers to evaluate EGFR, P-Akt, and HIF-1alpha expression. Significant correlation between P-Akt and HIF-1alpha (P=0.0006) and marginal correlation between EGFR and P-Akt (P=0.066) were found. P-Akt was shown to be a poor prognostic factor as to patients with lymph node involvement (P=0.031). PMID- 15363542 TI - Cancer chemopreventive activity of phenylpropanoids and phytoquinoids from Illicium plants. AB - In our joint project involving search of anti-tumor promoters from natural plant sources, six phenylpropanoids and seven phytoquinoids isolated from three Illicium plants (Illiciaceae) were tested for their inhibitory activities against Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O tetradecanoylphorbol-13-acetate in Raji cells. All tested compounds showed inhibitory activity against the EBV-EA activation even at 1 x 10mol ratio, and the inhibitory activity of their compounds was found to be more than that of beta carotene. Two phenylpropanoids having prenyl group, 4-allyl-2-methoxy-6-(3-methyl 2-butenyl)phenol (3) and 4-allyl-2,6-dimethoxy-3-(3-methyl-2-butenyl)phenol (4), showed more potent activities as anti-tumor promoters (IC50 224 and 217 mol ratio/TPA, respectively). The presence of a prenyl moiety in the phenylpropanoids plays an important role in anti-tumor promoting activity as xanthone, coumarin and flavonoid previously reported. This investigation indicated that prenylated phenylpropanoids might be valuable as potential cancer chemopreventive agents. PMID- 15363543 TI - Erythropoietin modulates the anticancer activity of chemotherapeutic drugs in a murine lung cancer model. AB - In this study, we assessed the ability of erythropoietin (EPO) to synergize with various chemotherapeutic agents and suppress the growth and metastasis of solid tumors. Animals were inoculated with Lewis lung carcinoma (LLC) cells and treated with EPO alone, the designated chemotherapeutic drug (cisplatin, mitomycin C or cyclophoshamide) alone, or EPO and the drug. Tumor volume was monitored daily. Thirteen days following cell injection, tumor mass was determined. In addition, the number of the metastatic foci in the lungs was determined. Cisplatin alone was capable of inducing a 7-fold decrease in final tumor volume compared to tumor bearing animals injected with saline. However, when EPO was combined with cisplatin, the animals experienced an 11-fold reduction in final tumor volume compared to saline-injected animals (P<0.001). A 2.5-fold reduction in tumor mass was observed in animals treated with cisplatin, compared to the saline-injected groups. Furthermore, injections of EPO and cisplatin induced a 4-fold reduction in tumor mass (P<0.001). Blood analysis indicated that a significant increase of more than 30% in WBC was found in animals injected concurrently with cisplatin and EPO, as compared to saline-injected mice (P<0.03). When EPO and mitomycin C were injected together, tumor mass was further reduced by 14% compared to that seen in mice treated with mitomycin C alone. However, this difference was not statistically significant. We conclude from this study that EPO can synergize with chemotherapeutic agents to further suppress the growth of tumors. The level of synergism is drug related. PMID- 15363544 TI - An in vitro short time-high dose drug exposure assay for predicting 5FU resistance of colorectal cancer. AB - The goal of this study was to develop a simple and rapid in vitro drug resistance assay to ascertain the effectiveness of 5-fluorouracil (5-FU) for the individual therapy of colorectal cancer. Colorectal cancer cells were isolated from tumor specimens and, after 4h exposure to high doses of 5-FU cell viability was measured with an ATP assay. The average IC50 concentration for 5-FU was calculated as 4000 microg/ml from 35 patients' tumors. The tumor cells were defined as extreme drug resistance with a survival rate 1 standard deviation (SD) over IC50, low drug resistance (LDR) with a survival rate 1 SD below IC50, and intermediate drug resistance (IDR) with survival rate between these two. The drug resistant assay for 102 patients' cancer cells showed that the proportion of patients with LDR to 5-fluorouracil was 19%. The in vitro drug resistance of the cancer cells was not correlated with cancer stages or by patient sex or age. However, most mucinous and poor differentiated cancer cells showed extreme or IDR. The in vitro ATP assay values for 25 Duke's D patients receiving postoperative 5-FU chemotherapy were comparable with clinical postchemotherapy responses. The sensitivity and specificity of the assay were 100 and 95%, respectively. This short time-high dose drug exposure assay may serve as an aid to improve 5-FU treatment for individual chemotherapy. PMID- 15363545 TI - Catechol-O-methyltransferase low activity genotype (COMTLL) is associated with low levels of COMT protein in human hepatocytes. AB - A single nucleotide polymorphism in the COMT (catechol-O-methyltransferase) gene that alters the amino acid sequence at codon 108 of S-COMT from val to met (val108met polymorphism) has been associated with a number of diseases and neuropsychiatric disorders. Several studies have shown that the met108 allele (COMTL) is associated with three to four-fold lower levels of COMT activity, compared to the val108 allele (COMTH), in extracts of human erythrocytes, liver and kidney tissue. We hypothesized that the differences in COMT activity observed in these studies were due to differing levels of COMT protein in cells and tissues with varying COMT genotypes. In order to address this, we obtained hepatocytes from 31 Caucasian female donors and determined their COMT genotype, COMT activity and COMT protein levels. We found that both cytosolic COMT activity and cytosolic COMT protein levels are lower in hepatocytes from COMTLL individuals, and that COMT activity levels correlate with COMT protein levels. Therefore, lower COMT activity seen in tissues and cells with the COMTLL genotype is likely due to lower COMT protein levels compared with tissues and cells from COMTHH individuals. PMID- 15363546 TI - CYP1A1 and GSTM1 polymorphisms in relation to lung cancer risk in Chinese women. AB - We examined CYP1A1 (I462V) and GSTM1 null polymorphisms in 200 female cases and 144 female controls selected from a population-based case-control study of lung cancer conducted in northeast China, where the rates of lung cancer among Chinese women are especially high. The CYP1A1 codon 462 point mutation in exon 7 (I462V) causes an Ile-Val substitution near the heme binding site. This mutation correlates with inducibility of aryl hydrocarbon hydrolase (AHH) activity, which activates polycyclic aromatic hydrocarbons (PAHs) in tobacco smoke and in indoor air pollution from coal-burning stoves, a risk factor for lung cancer in this study population. We found that the CYP1A1 I462V genotype (combined ile/val and val/val) was significantly associated with lung cancer risk. The odds ratio (OR) was 2.5 (95% confidence interval [CI], 1.55-4.03) after adjustment for significant risk factors such as age, ever smoking status, family history of cancer, and eye irritation when cooking. The association was more pronounced among non-smokers (OR=3.67; 95% CI, 1.85-7.28) than among smokers (OR=1.74, 95% CI, 0.85-3.54). In contrast, we did not find a significant association with the GSTM1 null genotype. In summary, our case-control study of lung cancer among women in northeast China revealed an elevated risk associated with the CYP1A1 I462V genotype, but no interaction with smoking or indoor air pollution was found. PMID- 15363547 TI - Suppression of invasion in human U87 glioma cells by adenovirus-mediated co transfer of TIMP-2 and PTEN gene. AB - TIMPs and PTEN are known to be inhibitors of the invasive activities of malignant glioma. But there has been no literature reported concerning the effect of combined gene transfer of these two genes on invasiveness of glioma. This study was designed to evaluate the effect of adenovirus-mediated in vitro gene transfer of tissue inhibitor of metalloproteinases-2 (TIMP-2) and phosphatase and tensin homology deleted on chromosome ten (PTEN) on invasion of human U87 glioma cells. The mRNA and protein expressions of TIMP-2 and PTEN in U87 cells infected with AdTIMP-2 and AdPTEN were determined by RT-PCR and Western blot, respectively. The relative activity of MMP-2 and MMP-9 were measured by Gelatin zymogram and invasion of U87 in vitro were detected using Boyden chamber. The number of invasion cell of U87, U87 infected with Ad-gal, AdPTEN, AdTIMP-2 and AdPTEN/TIMP 2 was 55.63+/-13.27, 48.27+/-14.75, 35.27+/-10.94, 27.37+/-12.81, and 19.17+/ 5.45, respectively. In vitro invasiveness of glioma cells was significantly inhibited by infection with AdTIMP-2 and/or AdPTEN, which was not consistent with the change of MMPs activity. And in the combinated group, the inhibition effect was more remarkable than in single group. Our studies suggest that adenovirus mediated combined TIMP-2 and PTEN gene therapy is possibly useful for anti invasion therapy of malignant glioma. PMID- 15363548 TI - Growth inhibitory effects of trastuzumab and chemotherapeutic drugs in gastric cancer cell lines. AB - The various treatments for advanced gastric cancer have limitations and induce only marginal survival benefit. HER-2/neu protein is overexpressed in several types of human cancers and its amplification is associated with poor prognosis. Recombinant humanized anti-HER-2/neu antibody (trastuzumab) not only inhibits the proliferation of HER-2/neu overexpressing tumor cells but also augments the cytotoxicity of concomitant chemotherapeutic agents in metastatic breast cancer. In this study, we evaluated the growth inhibitory effects of trastuzumab in gastric cancer cells. HER-2/neu protein was evaluated by immunohistochemical analysis in seven gastric cancer cell lines. MTT assay was performed to evaluate the growth inhibitory effects of trastuzumab and three chemotherapeutic agents, doxorubicin, cisplatin and paclitaxel, both alone and in combinations. The changes of cell cycle after trastuzumab treatment were analyzed by flow cytometry. Four of the cell lines, YCC-2 with strong positivity of HER-2/neu expression, NCI-N87 with moderate positivity, YCC-3 with weak positivity, and SK BR-3 as a positive control, were selected. After in vitro MTT assay for 1-day and 5 consecutive days' treatment of trastuzumab at various concentrations, growth inhibition was not observed in any cancer cell lines. However, there was variable dose-dependent sensitivity to doxorubicin, cisplatin and paclitaxel. YCC-2 and SK BR-3 cancer cells were more sensitive to three chemotherapeutic drugs, constantly (P<0.05). The combination of 5 consecutive days' treatment of trastuzumab with 1 day doxorubicin treatment showed significant growth inhibition only in YCC-2 and NCI-N87 gastric cancer cells. After 1-day trastuzumab treatment, the S-phase fraction was decreased by 52 and 70% in YCC-2 and SK-BR-3, respectively. In conclusion, the expressions of HER-2/neu protein in gastric cancer cells are variable, and concomitant treatments of trastuzumab with doxorubicin increase cytotoxicity. This suggests that trastuzumab-based biologic therapy with chemotherapeutic agents can be applied in gastric cancer treatment. PMID- 15363549 TI - Reduced expression of connexin 31.1 in larynx cancer is not caused by GJB5 mutations. AB - Lack of regular cell-cell interaction is one major cause for neoplastic growth and metastasis. In head and neck squamous cell carcinomas a 10-fold down regulation of connexin31.1 (GJB5) as well as mutations in the TGF-beta-receptor II were reported. We performed mutation screenings in GJB5 and the TGF-beta receptor-II poly(10)adenine hot spot employing larynx cancer samples of 10 patients. Variable length of the TGF-beta-receptor-II adenine homopolymer in controls and tumours indicate a high slippage error rate of the DNA polymerases rendering mutational analyses inconsistent. Lack of GJB5 mutations in the entire tumour collection suggests that this gene is not primarily involved in laryngeal tumorigenesis. PMID- 15363550 TI - CXCR4-mediated adhesion and MMP-9 secretion in head and neck squamous cell carcinoma. AB - The chemokine CXCL12 (SDF-1) and its receptor, CXCR4, have been implicated in organ-specific metastases of several malignancies. Head and neck squamous cell carcinoma (HNSCC) predominantly metastasizes to lymph nodes, and recent evidence has shown that CXCL12 stimulates HNSCC migration. We explored the potential role of CXCR4 in mediating other metastatic processes in HNSCC cells. CXCR4 mRNA and cell-surface expression was assessed in HNSCC cell lines. CXCR4 mRNA expression was detected in five HNSCC cell lines. Cell-surface CXCR4 was also detected in each of the HNSCC cell lines and in resected HNSCC tissues. CXCL12 induced rapid intracellular calcium mobilization in a metastatic HNSCC cell line (HN), as well as rapid phosphorylation of ERK-1/2. HNSCC cell adhesion to fibronectin and collagen was increased by CXCL12 treatment, while the addition of an inhibitor of ERK-1/2 signaling, PD98059, reduced the effects of CXCL12. CXCL12 also increased the active matrix metalloproteinase (MMP)-9 secreted. Thus, HNSCC cells express functional CXCR4 receptors that induce rapid intracellular signaling upon binding to CXCL12. Such binding leads to increased HNSCC cell adhesion and MMP secretion, suggesting that CXCR4 may be a novel regulator of HNSCC metastatic processes. PMID- 15363551 TI - Autocrine erythropoietin signaling inhibits hypoxia-induced apoptosis in human breast carcinoma cells. AB - Disordered perfusion and the resulting hypoxia are important features conferring tumor heterogeneity, which may contribute to relapse. Hypoxic tumor cells have been associated with resistance both to radiation and to cytotoxic drugs. Hypoxia may also serve as a selection pressure in tumors by promoting apoptosis of some cells and expanding variants with decreased apoptotic potential, and thus play a role in the development of a more aggressive phenotype. Erythropoietin (Epo), induced by hypoxia, controls erythropoiesis and plays a role in protection of neurons from hypoxic damage. We have recently demonstrated hypoxia-stimulated expression of Epo and Epo receptor (EpoR) in human breast and cervix cancers, suggesting a role for autocrine Epo signaling in the hypoxic adaptations of carcinomas. In the current study we provide evidence that increased autocrine Epo signaling induced by moderate levels of hypoxia inhibits hypoxia-induced apoptosis and promotes survival in MCF-7 human breast cancer cells. The anti apoptotic effect of Epo correlates with upregulation of bcl-2 and bcl-XL, suggesting a mechanism similar to those described in hematopoietic cells. The resulting decreased apoptotic potential of hypoxic tumor cells may contribute to increased aggressiveness and therapy resistance of breast cancers. PMID- 15363552 TI - What are the oncological risks of minimal access surgery for the treatment of urinary tract cancer? AB - OBJECTIVES: A review of the oncological safety of minimal access surgery for the treatment of urinary tract cancers. The particular areas reviewed were port-site metastases, local tumour recurrence and long-term survival. METHODS: Review of the literature using Medline. RESULTS: There is a low rate of port-site metastases following laparoscopic surgery for urological malignancies, these are usually related to the stage and grade of the tumour. So far follow-up data shows that laparoscopic surgery for urological malignancy does not result in higher levels of local recurrence or shorter survival than open surgery. Percutaneous (PCN) and ureteroscopic (URS) resection of TCC of the upper urinary tract are acceptable forms of treatment for grade 1 and 2 TCCs even in patients with normal contralateral kidneys. However, for grade 3 TCC nephroureterectomy should be utilised because of increased risk of local recurrence (URS) and track seeding (PCN). CONCLUSIONS: Provided the principles of cancer surgery, combined with proper case selection are followed, minimal access surgery for urological cancer is safe and is rapidly emerging as the standard of care for many upper tract tumours. PMID- 15363553 TI - Myelomeningocele and female sexuality: an issue? AB - During the last 40 years more and more myelomeningocele (MMC) patients have survived to adulthood and since the 1980s there is growing interest in sexual functioning of these patients. However, most of what is known about the impact of MMC on sexual functioning pertains to males. We have reviewed the literature pertaining to female sexuality and MMC with regard to sexual development, activity, dysfunction, sexual knowledge and education, pregnancy and delivery, as well as future trends. PMID- 15363554 TI - Nephroptosis and nephropexy--hung up on the past? AB - The label of 'an ineffective treatment for an imaginary disease' has largely been successfully stuck to the condition of nephroptosis and its surgical management, nephropexy. As a result, younger urologists, especially in the US and UK, have little knowledge of the condition nor why such accusations were made. In this review we explore the historical background to this statement, including descriptions of some of the more exotic surgical procedures employed in the past and visit the diagnosis again in the setting of the modern era, armed with more sophisticated diagnostic tools and less invasive and thus morbid surgical techniques. We aim to demonstrate that with appropriate diagnostic criteria, the diagnosis of symptomatic nephroptosis can be reliably made and that laparoscopic techniques provide an excellent approach in the successful surgical management of this condition. PMID- 15363555 TI - Handling and pathology reporting of circumcision and penectomy specimens. PMID- 15363556 TI - Port site metastasis after laparoscopic nephro-ureterectomy for transitional cell carcinoma. PMID- 15363557 TI - Tumor seeding following percutaneous antegrade treatment of transitional cell carcinoma in the renal pelvis. PMID- 15363558 TI - Case selection for radical prostatectomy in the UK. AB - INTRODUCTION: Radical prostatectomy is an increasingly popular treatment option for clinically localised prostate cancer, yet PSA outcome figures are rare in the UK. This makes it difficult to establish appropriate criteria for case selection. We conducted an audit of PSA recurrence of 5 large centres in the south of England and investigated the use of pre-operative PSA to improve case selection and outcome. METHOD: 854 patients notes were audited for pre-operative staging parameters and follow-up data obtained. Patients with neoadjuvant and adjuvant treatment as well as patients with incomplete data and follow-up were excluded. RESULT: Median follow-up was 52 months for the remaining 663 patients. Median PSA was 10 ng/ml. A large improvement of PSA recurrence free survival rates was observed from 1988 to 1998 as a result of change in case selection and stage migration. Overall Kaplan-Meier PSA recurrence free survival probability at 1, 3, 5 and 8 years was 0.83, 0.69, 0.60 and 0.48, respectively. Five-year PSA recurrence free survival probability for PSA ranges <4 ng/ml, 4.1-10 ng/ml, 10.1 20 ng/ml and >20 ng/ml was 0.82, 0.73, 0.59 and 0.20, respectively (Wilcoxon, p < 0.0001). A simulation of biochemical recurrence free survival for patient cohorts with stepwise reduced inclusion PSAs suggests an improved outcome for patients with a pre-operative inclusion PSA of <12 ng/ml. Further reduction of the inclusion PSA does not improve outcome. CONCLUSION: Intermediate PSA recurrence free survival has improved over time in England. PSA recurrence free survival estimates are less optimistic compared to frequently quoted American figures. A reduced pre-operative PSA cut-off for case selection may be used to improve outcome. PMID- 15363559 TI - Can Power Doppler enhanced transrectal ultrasound guided biopsy improve prostate cancer detection on first and repeat prostate biopsy? AB - OBJECTIVE: To determine the utility of Power Doppler enhanced transrectal ultrasound (PD-TRUS) and its guided prostate biopsies in men with prostate specific antigen (PSA) levels between 2.5 and 10 ng/ml and to evaluate its impact on prostate cancer (PCa) detection in men undergoing first and repeat biopsies. METHODS: A total of 136 consecutive referred men with serum total PSA (Abbott Laboratories, Abbott Park, IL, USA) levels between 2.5 and 10 ng/ml (mean age 64 +/- 9 years, range 45-82) and a normal digital rectal examination were included. 101 underwent a first biopsy whereas 35 had repeat biopsy. Gray-scale transrectal ultrasound (TRUS), and PD-TRUS (B&K Medical, Denmark) were performed in lithotomy position before and during the biopsy procedure. Vascularity accumulation and perfusion characteristics were recorded and graded as normal or abnormal in the peripheral zone of the prostate. A Vienna-nomogram based biopsy regime was performed in all patients on first biopsy and a special biopsy regime on repeat biopsy plus additional biopsies from abnormal sites on PD-TRUS. RESULTS: Overall PCa detection rate was 34.7% and 25.7% and abnormal accumulation on PD-TRUS was identified in 42.3% and 48.6% on first and repeat biopsy, respectively. The PCa detection rate, on first and repeat biopsy in patients with and without PD-TRUS accumulation were 67.4% versus 10.3% (p < 0.001) and 47.05% versus 5.6% (p = 0.0049), respectively. PD-TRUS directed biopsies were positive in 5.7% and 11.1% on first and repeat biopsy whereas PCa detection using the routine prostate biopsy regime was 94.3% and 88.9% on first and repeat biopsy. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of PD-TRUS signal alone for PCa detection on first biopsy was 82.8%, 78.8%, 87.9% and 89.7%, respectively, and 88.8%, 68.0%, 47.0% and 94.4% on repeat biopsy, respectively. In comparison, the results PD-TRUS guided biopsies were 53.8%, 59.1%, 16.7%, and 89.5%, on first biopsy, respectively, and 20.0%, 13.3%, 23.5%, 11.1% on repeat biopsy, respectively. CONCLUSION: Negative PD-TRUS signal is able to exclude most of the patients without PCa in the PSA range of 2.5-10 ng/ml. As an additional tool at TRUS biopsy PD-TRUS has a high negative predictive value and may help to reduce the number of unnecessary biopsies. PMID- 15363560 TI - Lymphadenectomy with cystectomy: is it necessary and what is its extent? PMID- 15363561 TI - Tumor grade-dependent alterations in the protein kinase C isoform pattern in urinary bladder carcinomas. AB - OBJECTIVES: Members of the protein kinase C (PKC) isoenzyme family play central role in the tumorigenesis of several tissues. In this study our goal was to determine the possible alterations in the protein kinase C (PKC) isoform pattern in relation with the different tumor grade in human urinary bladder carcinomas. METHODS: Western blot analysis, followed by quantitative densitometry, was performed to define the expression of PKC isoforms in the epithelial tissue of human urinary bladder carcinomas with various tumor grades and in control samples. RESULTS: The human urinary bladder epithelium expressed five PKC isoforms (PKC alpha, beta, delta, zeta), the levels of which differentially altered as a function of tumor grade. Namely, whereas the expressions of PKC beta and delta decreased with increasing grade of the carcinomas, the levels of PKC alpha, and zeta showed opposite patterns of changes. CONCLUSIONS: These grade dependent alterations in the PKC isoform pattern strongly argue for the central yet antagonistic roles of certain members of the PKC system in malignant transformation of human urinary bladder epithelium. PMID- 15363562 TI - Immunocyt and the HA-HAase urine tests for the detection of bladder cancer: a side-by-side comparison. AB - OBJECTIVES: The reliable detection of bladder cancer from urine specimen remains an unsolved problem. Especially superficial bladder cancer can be missed with urine tests. We assessed the sensitivity and specificity of the commercial Immunocyt test in a side-by-side comparison with the HA-HAase urine test and cytology. The Immunocyt test measures the immunocytological expression of sulfated mucin-glycoproteins and glycosylated forms of the carcinoembryonic antigen in urine. With the HA-HAase urine test the level of hyaluronic acid (HA) and its degrading enzyme hyaluronidase (HAase) are measured in an ELISA-like test. METHODS: A total of 94 consecutive patients were studied and among these 30 patients had bladder cancer and 64 were controls. Among bladder cancer patients, there were 14 pTa, 9 pT1, 5 pT2 and 2 carcinoma in situ (CIS) transitional cell carcinoma of the bladder, respectively. The controls consisted of 55 patients with a history of bladder cancer but no evidence of tumor at the follow-up cystoscopy and 9 benign prostatic hyperplasia (BPH) patients. The 30 transitional cell cancer specimens had 4 (13%) grade 1 tumors, 15 (50%) grade 2 tumors and 11 (37%) grade 3 tumors. Sensitivity and specificity as well as the positive and negative predictive values of each test were evaluated. RESULTS: The sensitivity of the HA-HAase urine test (83.3%; 25/30) was significantly higher than the Immunocyt at 63.3% (19/30) (p = 0.038, McNemar test) and cytology (73%; p < 0.05). The specificity of the HA-HAase test (78.1%; 50/64), Immunocyt (75%; 48/64) and cytology (79.7%; 51/64) were comparable. The prevalence of bladder cancer in our study was 31%. The positive predictive value (PPV) of the HA-HAase test (64.1%) was significantly higher than the Immunocyt test (54.3%). The negative predictive value (NPV) of the HA-HAase test (90.9%) was also higher than the Immunocyt test (81.3%). The PPV and NPV values for cytology were 62.9% and 86.4%, respectively. False negative patients in the HA-HAase urine test were 5 pTa tumors (2 G1, 2 G2 and 1 G3). False negative patients in the Immunocyt test were 7 pTa tumors (1 G1 and 6 G2), 3 pT1 (2 G2, 1 G3) and 1 pT2 G3, respectively. CONCLUSIONS: The sensitivity of the HA-HAase urine test is significantly higher than that of the Immunocyt test to detect bladder cancer. Specificity, as well as the PPV and NPV of the HA-HAase test were higher than that of the Immunocyt test. With a prevalence of 31% bladder cancer patients in all hematuria patients studied, a typical distribution of patients in a urological clinic is presented. Longer follow up of the study patients will give more information on the value of these tests in the detection of bladder cancer. PMID- 15363563 TI - Prognostic value of the involvement of the urinary collecting system in renal cell carcinoma. AB - OBJECTIVES: The prognostic role of the invasion of the urinary collecting system (UCS) by renal cell carcinoma (RCC) has not attracted a notable amount of attention. The aim of this study was to investigate incidence and prognostic value of UCS involvement in RCC. MATERIAL AND METHODS: All pathological reports of radical nephrectomies performed in two centres of urology from November 1983 to December 1999 were reviewed in order to evaluate the invasion of the UCS (calices, renal pelvis, ureter). Patients were divided into two groups according to presence (Group 1) or absence (Group 2) of UCS invasion. The stage was determined according to the TNM 6th edition. Overall and cause-specific survival rates were evaluated. Univariate and multivariate analyses were performed. RESULTS: The evaluable specimens were 671 from the 735 examined; in 64 cases it was not possible to ascertain or to exclude UCS involvement. Invasion of the UCS was found in 59 cases (8.8%). Median follow-up was 59.0 months (range 0-216). Tumours invading the UCS were usually symptomatic, with high nuclear grade and predominantly high stage. At univariate analysis the 5 year overall and cause specific survival rates of tumours invading the UCS were significantly lower when compared to those without UCS invasion (42.8% versus 60.8% and 45.5% versus 64.7%, respectively). When groups were stratified, according to the pT category, the 5-year cause-specific survival rate was only significantly different for the pT2 category (33.3% versus 76.9%). At the multivariate analysis TNM staging, symptoms at diagnosis and tumour grade were the only independent prognostic factors. CONCLUSION: The invasion of the UCS by RCC is unusual, particularly in small tumours. UCS involvement does not represent an independent prognostic factor. However, in organ-confined tumours (i.e. pT2) UCS involvement has an influence on the prognosis and should be taken into account when planning adjuvant treatments and follow-up. PMID- 15363564 TI - Long term results of excision of small renal cancer surrounded by a minimal layer of grossly normal parenchyma: review of 94 cases. AB - OBJECTIVE: To assess safety and effectiveness of excision of small renal cancer. METHODS: We reviewed the records of 94 patients, who underwent, from 1992 to 2001, excision of renal tumor leaving around it a thin layer of grossly normal parenchyma and adjacent perinephric fat. This technique has been called enucleoresection, as it is not a simple enucleation but neither a conventional partial nephrectomy. Patients were followed up mean routine blood examination, ultrasound and computed tomography scan every 6 months for 2 years and annually thereafter. RESULTS: Median age was 63 years (35-76). Median tumour size was 2.1cm (1.1-4.5). Clamping of renal pedicle was performed in 54 cases. Major complications included 4 cases of haemorrhage; only 1 patient required surgical exploration. Pathological stage was pT1a in 87, pT1b in 4 and pT3a in 3 patients. Surgical margins were always negative. Median follow-up was 59 months (range 10 128). Eight patients died without evidence of tumour recurrence. One pT3a patient developed distant metastases and died 2 years after surgery. Five years survival rate was 95.7% (90/94 patients), cancer specific survival rate 98.9% (93/94) and disease free survival rate 98.9% (93/94). CONCLUSIONS: Enucleoresection of small renal tumors surrounded by a minimal layer of grossly normal renal parenchyma reproduces the results of partial and radical nephrectomy with minimal morbidity. PMID- 15363565 TI - What is the relationship between male sexual function and lower urinary tract symptoms (LUTS)? AB - INTRODUCTION: There is considerable current interest in the relationship between LUTS and sexual function in men, and in particular whether there is a causative relationship between them. This observational study, performed in a single centre, seeks to explore this area further. METHODS: 1,420 patients attending a prostate assessment clinic were assessed by International Prostate Symptoms Score (IPSS), BPH impact index (BPH-II), Urinary flow rate (Q(max)), post-void residual volume (PVR) and the O'Leary sexual questionnaire (which assesses erectile, ejaculatory and sexual drive). The results were analyzed using univariate and multivariate techniques. RESULTS: Following exclusion of those with incomplete data, those who were not sexually active, those with PSA >20 ng/ml and those who refused to fill in the questionnaire, there were 696 evaluable patients. Using univariate techniques, age, IPSS, Quality of life (QOL), BPH-II, Q(max) and PVR all correlated with differing domains of sexual function including erectile and ejaculatory function. Multiple regression analysis demonstrated that age was overwhelmingly the most important predictor of sexual function, but that BPH-II and QOL were also predictive for some domains, while the IPSS score was not. CONCLUSION: This study confirms that a significant proportion of men with LUTS have sexual dysfunction. The relationship is complex and differs according to the type of sexual dysfunction. PMID- 15363566 TI - Efficacy and safety of long-term treatment with the dual 5 alpha-reductase inhibitor dutasteride in men with symptomatic benign prostatic hyperplasia. AB - OBJECTIVES: Dutasteride, a dual inhibitor of Type 1 and Type 2 5 alpha-reductase, has been shown to improve disease measures in patients with symptomatic benign prostatic hyperplasia (BPH) in three randomised, placebo-controlled, large-scale, 2-year Phase III clinical studies. This paper reports the pooled results of a 2 year open-label extension of the three randomised studies assessing the long-term efficacy and safety of dutasteride. METHODS: Patients randomised to dutasteride or placebo in the double-blind portion of the Phase III studies were eligible for a 2-year open-label extension, where all patients received dutasteride 0.5mg daily (dutasteride/dutasteride [D/D] group and placebo/dutasteride [P/D group]). RESULTS: Significant improvements in AUA-SI score and Q(max) were observed from Month 24 to 48 in both study groups. At Month 48, patients in the D/D group had significantly greater improvements in AUA-SI score and Q(max), and significantly greater reductions in prostate volume, than those in the P/D group. Acute urinary retention and BPH-related surgery occurred in a small percentage of patients during the open-label phase. No new safety issues were noted with long-term therapy. Onset of new drug-related adverse events were reported most frequently at the start of therapy and declined over time in patients receiving dutasteride. CONCLUSIONS: Long-term treatment with dutasteride results in continuing improvements in urinary symptoms and flow rate, and further reductions in TPV, in men with symptomatic BPH. The reduction in risk of AUR and BPH-related surgery, seen in the double-blind phase, was durable over 4-year treatment. Dutasteride was also well tolerated in long-term use. PMID- 15363567 TI - Evaluation of the transurethral ethanol ablation of the prostate (TEAP) for symptomatic benign prostatic hyperplasia (BPH): a European multi-center evaluation. AB - OBJECTIVE: We prospectively conducted a European multi-center study to assess the safety and efficacy of injecting dehydrated ethanol using a specialized injection system for the treatment of BPH. METHODS: Patients with symptomatic BPH were enrolled and evaluated to undergo transurethral ethanol ablation of the prostate for their BPH condition. Procedures were performed using the ProstaJect device. Treatment dosages were based on prostate volume, prostatic urethral length and median lobe involvement. Follow-up evaluations were done at four days and one, three, six and 12 months. RESULTS: One-hundred fifteen symptomatic patients underwent the transurethral ethanol ablation procedure and ninety-four patients have been followed and evaluated for the entire 12-month post-treatment period. The average prostate volume was 45.9 g, and average ethanol injected was 14 ml. Post-operatively, 98% of patients voided spontaneously four days following treatment. Significant reduction in reported lower urinary tract symptoms was evidenced at the one-month follow-up visit and maintained through 12 months follow-up, with International Prostate Symptom (IPSS) and Quality of Life (QoL) scores decreased by more than 50%. Peak flow rates (Q(max)) improved by 35% by the three-month evaluation and these results were sustained through to 12-months follow-up. The average prostate volume reduction was 16%. Adverse events included discomfort or irritative voiding symptoms in 26% of patients, hematuria in 16%, with retrograde ejaculation, and erectile dysfunction reported in less than 3% of patients. The majority of these events required no intervention. Two patients experienced serious adverse events (bladder necrosis) and underwent open surgery that included a urinary diversion and a ureteral implantataion. During the one year follow- up, 7% of patients required a trans-urethral resection of prostate (TURP). CONCLUSIONS: This preliminary multi-center data, representing the largest reported cohort to date, suggests that TEAP may be considered an effective minimally invasive treatment option for lower urinary tract symptoms secondary to BPH. Analyses of safety lead to a procedure modification for needle placement more distal from the bladder neck. Objective reduction in symptoms was not correlated in prostate volume reduction suggesting a non-purely mechanical effect. PMID- 15363568 TI - Efficacy of sildenafil citrate in treatment of erectile dysfunction: effect of type 2 diabetes. AB - PURPOSE: To assess efficacy of sildenafil citrate in treatment of erectile dysfunction: effect of type 2 diabetes. MATERIALS AND METHODS: A total of 466 male patients with erectile dysfunction (ED) were enrolled in this study. Of them 382 were diabetic and 84 were non-diabetic. Patients were screened for ED using the erectile function domain of the International Index for Erectile Function (IIEF). Patients underwent routine laboratory investigations, in addition to total testosterone and prolactin assessment. To assess the effect of diabetes on efficacy of sildenafil, we compared the pre and post sildenafil responses to erectile function domain, Q3, Q4. Overall satisfaction and global efficacy question (GEQ) were also assessed. RESULTS: Mean age +/- S.D. was 53 +/- 8.4 and 49.7 +/- 10.6 years for patients with and without diabetes respectively. There were significant associations between increased severity of ED and longer duration, poor metabolic control and presence of more than one diabetes-related complication (p < 0.05 for each). Differences were significant between pre and post sildenafil administration regarding erectile function domain, Q3, Q4 (p < 0.05 for each). In the non-diabetic patients the GEQ and the overall satisfaction were significantly higher than in diabetics (p < 0.05 for each). Global efficacy question was significantly low in patients with fair and poor metabolic control, longer duration of diabetes, and patients with diabetic complications (p < 0.05 for each). CONCLUSIONS: Sildenafil is an effective treatment for diabetic patients with ED. Although the efficacy of sildenafil was negatively affected by factors as poor control and longer duration of diabetes and presence of more than one diabetes-related complication, however, the global efficacy and the overall patients' satisfaction were high. PMID- 15363570 TI - Training and general and financial conditions of European residents in urology: an international survey. AB - OBJECTIVE: To assess the training and the general and financial conditions of European residents in urology. METHODS: Between February and April 2004 a questionnaire was sent by e-mail to 351 residents in urology in 30 different European countries. The questionnaire was divided into four different sections concerning general and financial conditions, clinical and surgical practice, research activities and participation in scientific meetings. RESULTS: One hundred and one trainees from 30 European countries completed the questionnaire and returned it, a response rate of 28.8%. The results of greatest interest were that 30% of the trainees feel there are too many residents in their departments, 34% have a compulsory annual assessment and 83.2% have a final qualifying examination. The majority of trainees say their experience in minor surgery is at least fair, while 33% and 36% have poor or non-existent experience in major surgery and endourology respectively. Furthermore, half of the respondents do not have the support of a tutor in their clinical practice. The European Association of Urology (EAU) guidelines are frequently used and their implementation in clinical practice is advocated by 96% of the respondents. The vast majority of the respondents are planning to become Fellows of the European Board of Urology (FEBU) and members of the EAU. CONCLUSION: This survey shows that, even if the general characteristics of different training programmes seem to be similar, the products of these systems present various discrepancies in terms of surgical and clinical practice. PMID- 15363569 TI - On-demand treatment of premature ejaculation with clomipramine and paroxetine: a randomized, double-blind fixed-dose study with stopwatch assessment. AB - OBJECTIVE: To investigate the degree of ejaculation delay induced by on-demand treatment with 20 mg paroxetine and 25 mg clomipramine and to assess the type and severity of non-sexual side-effects of treatment at the day of and the day after treatment with these drugs. METHOD: A randomized, double-blind, fixed-dose, on demand study in 30 men with lifelong premature ejaculation was performed. During a 1-month baseline period and a 4-week drug treatment period patients assessed the intravaginal ejaculation latency time (IELT) at home with a stopwatch. Only men with an IELT <1 min were randomly assigned to drug treatment. Patients assessed the drug coitus interval time (DCIT) and used the UKU side effect scale questionnaire at baseline, the day of and the day after intercourse. RESULTS: On demand treatment with 25 mg clomipramine, with a mean DCIT of 5.14 h, led to a 4.05 (95%CI: 3.26-5.02) fold-increase of the IELT. On-demand treatment with 20 mg paroxetine, with a mean DCIT of 5.39 h, led to a 1.41 (95%CI: 1.22-1.63) fold increase of the IELT. Both drugs had a high incidence of non-sexual side effects at the coitus day and the next day. At the day of coitus paroxetine led to significant sleepiness and yawning compared to clomipramine. At the day after coitus clomipramine induced significant nausea compared to paroxetine. CONCLUSION: On-demand treatment with 25 mg clomipramine led to a clinical relevant ejaculation delay. In contrast, 20 mg paroxetine had no clinical relevant ejaculation delay in men with lifelong premature ejaculation with an IELT of less than 1 minute. Both drugs exert mostly mild yet annoying non-sexual side effects both at the coitus day and the next day. PMID- 15363571 TI - Management of acute epididymitis: are European guidelines being followed? AB - OBJECTIVE: Acute epididymitis is increasing in men aged 35 years or under due to sexually-transmitted Chlamydia trachomatis. This study examines whether Urological surgeons are following European guidelines for the management of acute epididymitis in these patients. METHODS: A postal questionnaire survey was conducted of specialists in Urology in two regions of the UK. RESULTS: Of 79 completed replies, 41 (52%) take a detailed sexual history but only 34 (43%) refer patients to a Genitourinary medicine clinic. Quinolones are the most commonly prescribed first-line antibiotic by 56 (71%) respondents, principally ciprofloxacin. CONCLUSIONS: The current management of acute epididymitis in young men must be improved. Ciprofloxacin is not the optimal antimicrobial for the treatment of urogenital chlamydial infection. We recommend that all such patients be referred to local Genitourinary medicine services for contact-tracing and treatment of their sexual partners. PMID- 15363572 TI - Unstented tubularized incised plate urethroplasty combined with foreskin reconstruction for distal hypospadias. AB - BACKGROUND AND AIM: Urethral stent has recently been proven to be unnecessary for normal healing in an animal model of tubularized incised plate (TIP) urethroplasty. We report our experience with unstented TIP repair combined with foreskin reconstruction for distal hypospadias in children. PATIENTS AND METHODS: We retrospectively reviewed the records of 162 children consecutively treated by TIP urethroplasty for a distal or mid-shaft hypospadias without urethral stent over a 6 years period. The mean age +/- SEM at surgery was 15.7 +/- 1.2 months. A foreskin reconstruction was performed with the hypospadias repair in 136 boys (84%). One hundred thirty one children (81%) underwent this surgery as an outpatient procedure. RESULTS: With a mean follow-up of 12.4 +/- 1.0 months, urethrocutaneous fistula was observed in 9 children (5.6%), and meatal stenosis in 4 (2.5%). Postoperative urinary retention requiring suprapubic catheter insertion was observed in 4 cases (2.5%) without later complications. Cutaneous dehiscence of the reconstructed foreskin occurred in 6 children (4.4%) and phimosis in 13 (9.5%). CONCLUSIONS: Absence of urethral stent after TIP urethroplasty for distal hypospadias repair does not seem to increase postoperative complication rate. Foreskin reconstruction in distal hypospadias surgery has an acceptable complication rate. PMID- 15363573 TI - Bladder cell culture on small intestinal submucosa as bioscaffold: experimental study on engineered urothelial grafts. AB - OBJECTIVES: To investigate the feasibility to perform primary urothelial cell culture using porcine small intestinal submucosa as a delivery scaffold both in vitro and after in vivo implantation in a rabbit model. MATERIALS AND METHODS: Bladder mucosa samples were aseptically obtained from a group of eight male rabbits. The mucosa was cut into fragments and placed on small intestinal submucosa matrices for selective urothelial cell culture. After complete in vitro epithelization the matrices were shaped into tubes and placed in the subcutaneous tissue and subdartos of donor rabbits. The pattern of cell growth and delivery was evaluated on retrieved grafts using histology and immunostaining at the end of the in vitro phase; then 5, 10 and 20 days after implantation. RESULTS: Histological and immunohistochemical analysis of the in vitro primary culture showed the acellular matrices covered with a thin uninterrupted monolayer of urothelial cells. The implants examined on the day 5 maintained the epithelial configuration of the cultured grafts in all samples retrieved. On the day 10 the urothelium showed increased thickness taking on a bilayer configuration. On day 20, all grafts presented the transitional cells arranged in a double layer closely resembling the natural urothelium. The immunostaining pattern displayed the maintaining of urothelial cell phenotype. No differences in epithelium growth and delivery were noted between the two sites of implantation. Five days after implantation, the histological analysis of small intestinal submucosa showed a medium degree tissue reaction with the presence of acute inflammatory cells. Angiogenesis was demonstrated by the development of several new vessels inside the matrix. After twenty days, small intestinal submucosa was gradually replaced with host tissue. CONCLUSION: The small intestinal submucosa proved to function as a means of delivering of autologous urothelial cells cultured in vitro. After ectopic in vivo implantation the bioscaffold maintained viability and growth of the surrounding cells until its degradation. PMID- 15363574 TI - Re: Winstanley AM, Mikuz G, Debruyne F, Schulman CC, Parkinson MC. Handling and reporting of biopsy and surgical specimens of testicular cancer. Eur Urol 2004;45:564-73. PMID- 15363576 TI - Re: Lang H, Lindner V, Letourneux H, Martin M, Saussine C, Jacqmin D. Prognostic value of microscopic venous invasion in renal cell carcinoma: long-term follow up. Eur Urol 2004;46:331-5. PMID- 15363577 TI - Mitochondrially-mediated toxicity of bile acids. AB - In the healthy hepatocyte, uptake of bile acids across the basolateral membrane and export via the canalicular export pump, are tightly coupled. Impairment of bile formation or excretion results in cholestasis, characterized by accumulation of bile acids in systemic blood and within the hepatocyte. When the concentration of bile acids exceeds the binding capacity of the binding protein located in the cytosol of the hepatocyte, bile acids induce apoptosis and necrosis, by damage to mitochondria. Mitochondria play a central role on the toxicity of bile acids. In this article, we review the published literature regarding bile acid effects on cell function, especially at the mitochondrial level. In patients with cholestatic liver disease, the extent of hepatocyte damage caused by intracellular accumulation of bile acids appears to be delayed by ingesting a hydrophilic bile acid. However, its effects on disease progression are not completely clarified. Therefore, identification of the mechanisms of cell injury will be of clinical utility, helping in the development of new therapeutic strategies. The goal of this review is to include a fresh consideration of all possible targets and integrating pathways that are involved in cholestasis, as well as in the benefits of bile acid therapy. PMID- 15363578 TI - Lymphocyte DNA damage in rats exposed to pyrethroids: effect of supplementation with Vitamins E and C. AB - Pesticides have been considered potential chemical mutagens. In fact, some studies show that various agrochemical ingredients possess mutagenic properties inducing mutations, chromosomal alterations or DNA damage. Experimental evidence shows a marked correlation between mutagenicity and carcinogenicity and indicates that short-term mutagenicity tests are useful for predicting carcinogenicity. The present study on rat exposed to two pyrethroids, cypermethrin and permethrin, showed different lymphocyte DNA damage depending on the type of pyrethroid, the dose, and the period of treatment. Data obtained from comet assay showed that oral treatment with 150 mg/kg body weight/day of permethrin (corresponding to 1/10 of LD50) for 60 days, induced a significant increase in all comet parameters. No lymphocyte DNA damage was measured after treatment with 25 mg/kg body weight/day of cypermethrin (corresponding to 1/10 of LD50) for the same period. A higher dose of permethrin (300 mg/kg body weight/day), for a shorter period (22 days), did not induce lymphocyte DNA damage, while supplementation with 200 mg/kg of Vitamins E and C protected erythrocytes against plasma membrane lipids peroxidation. Moreover, treatment with Vitamins E and C maintained the activity of glutathione peroxidase, which was reduced in the presence of permethrin, and reduced the osmotic fragility, which had increased following permethrin treatment. PMID- 15363579 TI - Toxicity classification and evaluation of four pharmaceuticals classes: antibiotics, antineoplastics, cardiovascular, and sex hormones. AB - Four different classes of environmental concern are quantitatively and qualitatively assessed for environmental hazards; antibiotics (n = 226), antineoplastics (n = 81), cardiovascular (n = 272), and sex hormones (n = 92). These along with an ECOSAR scan of all pharmaceuticals (n = 2848) were then classified according to the OECD aquatic toxicity classification system. The predicted species susceptibility is: daphnid > fish > algae, and the predicted rank order of relative toxicity: sex hormones > cardiovascular = antibiotics > antineoplastics (Table 1). Generally, a relatively large proportion (1/3) of all pharmaceuticals are potentially very toxic to aquatic organisms (Table 2). The qualitative risk assessment ranking relative to probability and potential severity for human and environmental health effects is: antibiotics > sex hormones > cardiovascular > antineoplastics. (Q)SARs and pharmacodynamic information should be used to prioritize and steer experimental risk assessments of pharmaceuticals, and potentially, also be used in new drug discovery optimizing efficacy and in minimising environmental hazards of new products. Nuclear receptors are relatively well conserved in evolution. Currently, antibacterial resistance represents the most significant human health hazard, and potentially the largest non-target organism hazard is sex hormones acting as endocrine modulators in wildlife. Data for the individual compounds are accessible via. PMID- 15363580 TI - PPARalpha activators down-regulate CYP2C7, a retinoic acid and testosterone hydroxylase. AB - Peroxisome proliferators (PP) are a large class of structurally diverse chemicals that mediate their effects in the liver mainly through the peroxisome proliferator-activated receptor alpha (PPARalpha). Exposure to PP results in down regulation of CYP2C family members under control of growth hormone and sex steroids including CYP2C11 and CYP2C12. We hypothesized that PP exposure would also lead to similar changes in CYP2C7, a retinoic acid and testosterone hydroxylase. CYP2C7 gene expression was dramatically down-regulated in the livers of rats treated for 13 weeks by WY-14,643 (WY; 500 ppm) or gemfibrozil (GEM; 8000 ppm). In the same tissues, exposure to WY and GEM and to a lesser extent di-n butyl phthalate (20,000 ppm) led to decreases in CYP2C7 protein levels in both male and female rats. An examination of the time and dose dependence of CYP2C7 protein changes after PP exposure revealed that CYP2C7 was more sensitive to compound exposure compared to other CYP2C family members. Protein expression was decreased after 1, 5 and 13 weeks of PP treatment. CYP2C7 protein expression was completely abolished at 5 ppm WY, the lowest dose tested. GEM and DBP exhibited dose-dependent decreases in CYP2C7 protein expression, becoming significant at 1000 ppm or 5000 ppm and above, respectively. These results show that PP exposure leads to changes in CYP2C7 mRNA and protein levels. Thus, in addition to known effects on steroid metabolism, exposure to PP may alter retinoic acid metabolism. PMID- 15363581 TI - Genotoxic effects on human spermatozoa among pesticide factory workers exposed to fenvalerate. AB - Fenvalerate, a synthetic pyrethroid insecticide, is widely produced and used worldwide. To explore fenvalerate-induced genotoxic effects, particularly numerical chromosome aberration (CA), we firstly examined conventional semen parameters, the progression and motion parameters of the spermatozoa among 12 fenvalerate-exposed workers and 30 donors of the internal and external control groups. Then numerical CA of chromosome X, Y and 18 were investigated by multicolor fluorescence in situ hybridization (FISH). The results showed the significant differences in the percentage of sperm abnormality between fenvalerate-exposed group and the external control group (P = 0.024). In aneuploid parameters, the frequency (mean +/- S.D.) of sex chromosome disomy was 0.742 +/- 0.131% in fenvalerate-exposed group, which was significantly higher than those in the internal (0.563 +/- 0.135%) and external control group (0.386 +/- 0.140%) (P < 0.01), and the frequency of chromosome 18 disomy in fenvalerate exposed group (0.326 +/- 0.069%) was significantly higher than those in the internal and external control groups (0.195 +/- 0.094% and 0.124 +/- 0.068%), respectively (P < 0.01). We also found the nullisomies of sex chromosomes and chromosome 18 were significantly higher than those in the external control group and two control groups, respectively (P < 0.01). The frequencies of aneuploidy and numerical CA we detected also showed significant differences between exposed group and control groups (P < 0.05 and/or P < 0.01). Moreover, we found the positive correlation not only between nullisomic frequencies of these chromosomes and numerical CA rate (r > 0.70, P < 0.01) but also between disomic frequency of sex chromosomes, aneuploidy rate and sperm abnormality in all donors (r = 0.530 and r = 0.536, P < 0.01). Our findings suggest that fenvalerate or its metabolites induced morphologic abnormality and genotoxic defects of spermatozoa among fenvalerate-exposed workers by causing numerical CA in spermatogenesis as a special and potential genotoxic agent. PMID- 15363582 TI - Alterations of FSH-stimulated progesterone production and calcium homeostasis in primarily cultured human luteinizing-granulosa cells induced by fenvalerate. AB - Fenvalerate, a synthetic pyrethroid, is widely used in agriculture and other domestic applications in China. Recently, Fenvalerate has been suspected to be one of the endocrine-disrupting chemicals (EDC). In this study, we investigated the effects of fenvalerate on follicle-stimulating hormone (FSH)-stimulated progesterone (P4) production by human ovarian luteinizing-granulosa cells (hGLCs). After 24 h incubation, fenvalerate inhibited FSH-stimulated P4 production. At the same time, FSH-stimulated cAMP also decreased. Due to calcium and Ca2+ -calmodulin (CaM) system involving gonadotropin-stimulated steroidogenesis by granulosa cells, we then evaluated the effects of fenvalerate on trifluoperazine (TFP)- and verapamil-driven FSH-stimulated P4 production. The results showed that calcium or calmodulin might play a role in fenvalerate induced alterations in FSH-stimulated P4 biosynthesis. Then, the effects of fenvalerate on calcium homeostasis in hGLCs were studied. The result showed that 5 microM fenvalerate induced a slow increase in [Ca2+]i in hGLCs by using a fluorescent Ca2+ indicator fluo-3/AM. The changes in total concentration of CaM in hGLCs induced by fenvalerate were evaluated by a method of immunofluorescence. There is a significant increase in all treated groups. In summary, fenvalerate could inhibit FSH-stimulated P4 production. Also, fenvalerate interferes with calcium homeostasis in hGLCs. The effects of fenvalerate on FSH-stimulated ovarian steroidogenesis may be mediated partly through calcium signal. PMID- 15363583 TI - H2S cytotoxicity mechanism involves reactive oxygen species formation and mitochondrial depolarisation. AB - A number of scavengers of reactive oxygen species (ROS) were found to be protective against cell death induced by hydrogen sulfide (H2S) in isolated hepatocytes. The H2O2 scavengers alpha-ketoglutarate and pyruvate, which also act as energy substrate metabolites, were more protective against H2S toxicity than lactate which is only an energy substrate metabolite. All of these results suggest that H2S toxicity is dependent on ROS production. We measured ROS formation directly in hepatocytes using the fluorogenic dichlorofluorescin method. H2S-induced ROS formation was dose dependent and pyruvate inhibited this ROS production. Non-toxic concentrations of H2S enhanced the cytotoxicity of H2O2 generated by glucose/glucose oxidase, which was inhibited by CYP450 inibitors. Furthermore, hepatocyte ROS formation induced by H2S was decreased by CYP450 inhibitors cimetidine and benzylimidazole. These results suggest that CYP450 dependant metabolism of H2S is responsible for inducing ROS production. H2S induced cytotoxicity was preceded by mitochondrial depolarization as measured by rhodamine 123 fluorescence. Mitochondrial depolarization induced by H2S was prevented by zinc, methionine and pyruvate all of which decreased H2S-induced cell death. Treatment of H2S poisoning may benefit from interventions aimed at minimizing ROS-induced damage and reducing mitochondrial damage. PMID- 15363584 TI - The effects of zinc deficiency and supplementation on lipid peroxidation in bone tissue of ovariectomized rats. AB - This study aims at investigating how zinc deficiency and zinc application affect lipid peroxidation in bone tissue of ovariectomized rats. The study included 40 adult female rats of Sprague-Dawley species. Group 1 (n = 10): control group were fed with normal diet. Group 2 (n = 10): the group fed with normal diet after being ovariectomized. Group 3 (n = 10): the group fed with zinc-deficient diet for 6 weeks after ovariectomy. Group 4 (n = 10): the group which was given intraperitoneal zinc (3 mg/kg day zinc) in addition to normal diet for 6 weeks after ovariectomy. Malondialdehyde (MDA) and glutathione (GSH) levels were determined in erythrocyte, plasma and bone tissue. Group 3 had the highest plasma MDA levels compared to Groups 1, 2 and 4 (P < 0.05). These values were higher in Group 2 than in Groups 1 and 4 (P < 0.05). Bone and plasma MDA levels in Groups 1 and 4 were not different. Bone and erythrocyte GSH levels in Group 4 were higher than those in all other groups (P < 0.05). The lowest levels of bone and erythrocyte GSH levels were observed in Group 3 (P < 0.05). These values were higher in Group 2 when compared to those in Groups 1 and 3 (P < 0.05). This study demonstrate that zinc deficiency increased bone tissue damage in ovariectomized rats and that zinc supplementation prevented this damage. PMID- 15363586 TI - Influence of a combination of two tetrachlorobiphenyl congeners (PCB 47; PCB 77) on thyroid status, choline acetyltransferase (ChAT) activity, and short- and long term memory in 30-day-old Sprague-Dawley rats. AB - The important role of thyroid hormones in growth and development, maintenance of body temperature, digestion, cardiac function, and normal brain development can be disrupted by environmental contaminants like polychlorinated biphenyls (PCB). Polychlorinated biphenyls are environmental contaminants that are widespread, persistent, lipophilic, and bioaccumulate through food webs, concentrating in adipose tissue. Placental and lactational PCB exposure of offspring causes metabolic and endocrine disruptions including hypothyroxinemia, spatial learning and memory deficits, neurochemical and neurobehavioral alterations, and reproductive problems. Previous studies in our lab using the individual congeners PCB 47 (2,2',4,4'-tetrachlorobiphenyl, ortho-substituted) and PCB 77 (3,3',4,4' tetrachlorobiphenyl, non-ortho-substituted) have demonstrated alterations in thyroid hormone levels, alterations in brain choline acetyltransferase (ChAT) activity, and spatial learning deficits. In the present study, pregnant Sprague Dawley rats were fed a diet with or without a mixture of PCB 47/77 at 1.25 ppm, 12.5 ppm or 25.0 ppm (w/w). Rat pups were swum in the Morris water maze four times a day on days 21-29 in order for the animals to learn the position of a submerged fixed platform. A probe test was run on day 24 (30 min after last swim) for short-term memory, and on day 29 (24 h after the last swim) for long-term memory after removal of the platform. Time spent in the quadrant previously containing the platform was recorded. Rats were decapitated on day 30, serum collected and frozen at -20 degrees. ChAT activity was measured radiometrically in basal forebrain and hippocampus. All PCB-treated animals experienced a depression in both triiodothyronine (T3) and thyroxine (T4). The present study found that all doses of PCB depressed ChAT activity in hippocampus with no significant alteration in the basal forebrain. In PCB-treated animals, short-term memory showed a trend toward improvement and long-term memory toward depression, but these trends were not significant. The consequences likely stem from endocrine disruption, especially with regard to the thyroid. PMID- 15363585 TI - Role of the peroxisome proliferator-activated receptor alpha (PPARalpha) in responses to trichloroethylene and metabolites, trichloroacetate and dichloroacetate in mouse liver. AB - Trichloroethylene (TCE) is an industrial solvent and a widespread environmental contaminant. Induction of liver cancer in mice by TCE is thought to be mediated by two carcinogenic metabolites, dichloroacetate (DCA) and trichloroacetate (TCA). TCE is considered to be a relatively weak peroxisome proliferator (PP), a group of rodent hepatocarcinogens that cause adaptive responses in liver through the PP-activated receptor alpha (PPARalpha). The objectives of this study were to determine whether effects of TCE, TCA and DCA in the liver associated with carcinogenesis are mediated by PPARalpha. Male wild-type and PPARalpha-null mice were given TCE by gavage for 3 days or 3 weeks; TCA or DCA were given in the drinking water for 1 week. Increases in relative liver and kidney weights by TCE were dependent on PPARalpha whereas liver weight increases by DCA were PPARalpha independent. Dose-dependent increases in hepatocyte proliferation observed in wild-type mice after TCE exposure as determined by BrdU-labeling of hepatocytes were PPARalpha-dependent. Transcript profiling using macroarrays containing approximately 1200 genes showed that 93% (40 out of 43) of all expression changes observed in wild-type mice upon TCE exposure were dependent on PPARalpha and included known targets of PP (Cyp4a12, epidermal growth factor receptor) and additional genes involved in cell growth. Increases in enzymes that catalyze beta and omega-oxidation of fatty acids were dependent on PPARalpha after exposure to TCE, TCA or DCA. TCE altered a unique set of genes in the livers of PPARalpha null mice compared to wild-type mice including those that respond to different forms of stress. These data support the hypothesis that PPARalpha plays a dominant role in mediating the effects associated with hepatocarcinogenesis upon TCE exposure. PMID- 15363587 TI - The shark bile salt 5 beta-scymnol abates acetaminophen toxicity, but not covalent binding. AB - Acetaminophen (APAP) toxicity involves both arylative and oxidative mechanisms. The shark bile salt, 5 beta-scymnol (5beta-S), has been demonstrated to act as an antioxidant and free radical scavenger in vitro. To determine if 5beta-S protects against either APAP-induced hepatic or renal toxicity, 3-4-month-old male Swiss Laca mice were given APAP (500 mg/kg), and 5beta-S (100 mg/kg) was given at 0 and 2 h after APAP. Plasma SDH at 12 h after APAP alone was 1630 U/l and BUN was 19 mg/dl versus 20 U/l and 10 mg/dl, respectively, in controls. Either simultaneous or 2 h delayed treatment with 5beta-S significantly decreased the APAP-induced SDH increase while only the simultaneous pretreatment prevented the BUN elevation. 5beta-S alone did not increase liver glutathione content. Western analysis of APAP covalent binding using anti-APAP antibodies indicated the 5beta S did not alter protein arylation either qualitatively or quantitatively. These results suggest that 5beta-S treatment did not impair APAP activation and are consistent with 5beta-S protection that likely results from its antioxidant activity. PMID- 15363588 TI - Dehydrocrotonin and its derivative, dimethylamide-crotonin induce apoptosis with lipid peroxidation and activation of caspases-2, -6 and -9 in human leukemic cells HL60. AB - A variety of stimuli can induce cells to undergo apoptosis, with one of the most reproducible inducers being mild oxidative stress following exposure to anticancer agents. Apoptosis involves events mediated by cysteine proteases (caspases) that are classified as initiators (-8, -9 and -12) or executors (-2, 3, -6 and -7). In this study, we examined the mechanisms of apoptosis induced by dehydrocrotonin (DHC), a diterpene lactone isolated from the Amazonian plant Croton cajucara, and its synthetic derivative, dimethylamide-crotonin (DCR), in human HL60 promyelocytic leukemia cells. Flow cytometric analysis of HL60 cells after treatment for 72 h showed that DCR- and DHC-induced apoptosis, with maximum cell death at a concentration of 250 microM for both compounds. DCR and DHC were effective in triggering the activation of caspases-2, -6 and -9. The level of reduced glutathione (GSH) decreased, whereas there was an increase in thiobarbituric acid-reactive substance (TBARS) production and in mitochondrial swelling. These effects on mitochondrial swelling, GSH content and lipid peroxidation were abolished by cyclosporine A, an inhibitor of the membrane permeability transition. The cytotoxicity of DHC and DCR was prevented by a high concentration of GSH (15 mM) in the culture medium. These results indicate that DCR and DHC produced apoptosis partly by oxidative stress-induced lipid peroxidation, which triggered the caspase cascade, that lead to apoptotic cell death in HL60 cells. Based on the pattern of caspase activation, on the increase in mitochondrial swelling and on the inhibitory action of cyclosporine A, we conclude that DCR and DHC triggered apoptosis in HL60 cells probably through cytochrome c release and apoptosome formation. PMID- 15363589 TI - The subchronic toxicity of hydroxysafflor yellow A of 90 days repeatedly intraperitoneal injections in rats. AB - Hydroxysafflor yellow A (HSYA) is the main segment of the safflower yellow pigments, and its second clinical study has been approved by the China SFDA for the treatment of brain blood vessel disease. Present studies were conducted to demonstrate the subchronic toxicity of HSYA. Sprague-Dawley (SD) rats were tested by daily intraperitoneal (IP) injection of HSYA at the doses of 180, 60, 20 mg/kg for 90 days. HSYA at high dose of 180 mg/kg and at middle dose of 60 mg/kg (90 day daily injection) induced a prolonged blood coagulation time without influencing the normal blood coagulation process. No animal died from hemorrhaging. The prolonged blood coagulation time was recovered to a normal level on the 28th day after withdrawing the drug. Kidney injury, including round tubular figures and a breaking-off of the tubular epithelium in histological slices under the microscope scenic, was only observed in the rats given by HSYA at 180 mg/kg. HSYA at dose of 180 mg/kg also increases the liver index without an obvious pathological change in liver histological analysis. There was no other organ injury found in this study. In conclusion, 90 days of HSYA treatment at a dose of 180 mg/kg, not at a dose of 60 mg/kg, induces a slight nephrotoxicity. PMID- 15363590 TI - Long-term cadmium exposure accelerates age-related mitochondrial changes in renal epithelial cells. AB - Long-term cadmium exposure leads to mitochondrial dysfunction in the proximal tubular epithelial cells. Mitochondrial DNA deletion may contribute to the pathogenesis of cadmium-induced nephropathy. The aim of our study is to clarify the accumulation of mitochondrial DNA deletion and mitochondrial dysfunction in the renal cortex of rats injected three times/week with 1 ml of 1 mM CdCl2 or saline for 80 weeks. After 40-week cadmium injection, mitochondrial number diminished, and cadmium in the renal cortex reached a saturation level. At this time interval, nearly 30% of cadmium in the whole cell fraction was found in the mitochondria. Cytochrome c oxidase (COX) activity in the proximal tubular epithelial cells decreased after 40-week exposure of cadmium. Oxidized phosphatidylcholine (oxPC) started to accumulate in the cytochrome c-positive mitochondria in some tubular epithelial cells after 80-week exposure. After 40 weeks, accumulation of the 4834-bp deletion in mitochondrial DNA was evident in both control and cadmium-treated groups. However, the amount of accumulated mitochondrial DNA deletion tended to increase after 40-week exposure, and was significantly greater after 80 weeks of exposure, compared to the control. Our results indicate that long-term cadmium exposure in rats accelerates accumulation of 4834-bp mitochondrial DNA deletions and impairment of mitochondrial function associated with accumulation of oxidized product. PMID- 15363591 TI - A trivalent dimethylarsenic compound, dimethylarsine iodide, induces cellular transformation, aneuploidy, centrosome abnormality and multipolar spindle formation in Syrian hamster embryo cells. AB - The abilities of dimethylarsine iodide (DMI), a model compound of trivalent dimethylarsenicals, to induce cellular transformation, aneuploidy, centrosome abnormality, and multipolar spindle formations were investigated using the Syrian hamster embryo (SHE) cell model. Cellular growth was decreased in a concentration dependent manner by treatment with DMI at concentrations over 0.1 microM. Treatment with DMI at concentrations from 0.1 to 1.0 microM induced morphological transformation in SHE cells. The transforming activity of DMI, determined by the frequency of morphologically transformed colonies, was approximately 30 times higher than that induced by treatment with the same concentration of sodium arsenite. Flow cytometry suggested an increase in the aneuploid population caused by DMI, as shown by the appearance of hypo-2N, hypo-4N and hypo-8N. DMI also caused abnormal staining of gamma-tubulin, indicating loss of centrosome integrity and a resultant induction of multipolar spindles in mitotic cells. Mitotic cells with centrosomes that coalesced partly at the cell periphery, not the cell center, were detected as early changes that resulted in multipolar spindles. These findings indicate that DMI has transforming activity in SHE cells. Moreover, the results suggest the importance of centrosome abnormalities as a causal change of DMI-induced aneuploidy. PMID- 15363592 TI - Lack of in vitro protection by a common sunscreen ingredient on UVA-induced cytotoxicity in keratinocytes. AB - As an extension of our previous investigations on sunscreen ingredients, the present work was aimed at assessing the possible protective effects of a common UVA-absorbing agent, Parsol 1789 (4-tert-butyl-4'-methoxydibenzoylmethane) in contact with human keratinocytes under UVA illumination. Cell viability was evaluated by determining lactate dehydrogenase (LDH) release, uptake of propidium iodide and fluorescein diacetate, total protein content and percentage of cell detachment. Apoptosis was detected by recognition of translocated phosphatidylserine using annexin V-FITC uptake. Oxidative stress was evaluated through the carboxy-H2DCFDA assay while the total oxyradical scavenging capacity (TOSC) assay was used for determining the total antioxidant capacity level in these cells. Lipid peroxidation was also assessed by checking hydroperoxide (HP) levels. The results obtained show that UVA exposure induces significant cell mortality, decrease in protein concentration, release of LDH, increase in apoptosis, oxidative stress and lipid peroxidation with a concomitant reduction in the response of the antioxidant cellular defense system. The presence of 10 microM Parsol 1789 did not minimize these UVA-induced effects, on the contrary, for some parameters measured such as lipid hydroperoxides, there was a significant enhancement. Furthermore, the presence of glutathione (GSH) alone decreased the level of ROS and lipid hydroperoxides, but in combination with Parsol 1789, this protective effect was reduced. The overall results indicate that the compound does not protect these cells from UVA exposure under our experimental conditions confirming previous findings on the lack of photoprotective efficiency of this sunscreen in contact with biologically relevant molecules. However, the biological role and significance of these results to the consequences of sunscreen use in humans are not known, hence extrapolation from laboratory experiments must be done with caution. PMID- 15363593 TI - Contributory role of 5-lipoxygenase and its association with angiogenesis in the promotion of inflammation-associated colonic tumorigenesis by cigarette smoking. AB - Our previous study shows that cigarette smoking can promote inflammation associated adenoma formation in the mouse colon, but the underlying mechanism remains unknown. Several studies suggest that there is a link between 5 lipoxygenase (5-LOX) and carcinogenesis in humans and animals. In the present study, we aims to investigate whether the promoting action of cigarette smoke on inflammation-associated colon cancer formation is associated with 5-LOX activation in mice. Results showed that exposure to the mainstream smoke of unfiltered cigarettes enhanced the 5-LOX protein expression in the inflammation associated colonic adenomas. It was accompanied with an up-regulation of matrix metalloproteinase-2 (MMP-2) and vascular endothelial growth factor (VEGF). Both are the key angiogenic factors for tumorigenesis. 5-LOX inhibitors decreased the incidence of colonic adenoma formation and reduced angiogenesis, MMP-2 activity and VEGF protein expression in the colons of these animals. Taken together, these results strongly suggest that cigarette smoke can induce 5-LOX expression which plays an important role in activation of MMP-2 and VEGF to induce angiogenic process and promotion of inflammation-associated adenoma formation in mice. PMID- 15363594 TI - Strain differences of cadmium-induced hepatotoxicity in Wistar-Imamichi and Fischer 344 rats: involvement of cadmium accumulation. AB - We previously reported that Wistar-Imamichi (WI) rats have a strong resistance to cadmium (Cd)-induced lethality compared to other strains such as Fischer 344 (Fischer) rats. The present study was designed to establish biochemical and histological differences in Cd toxicity in WI and Fischer rats, and to clarify the mechanistic basis of these strain differences. A single Cd (4.5 mg/kg, s.c.) treatment caused a significant increase in serum alanine aminotransferase activity, indicative of hepatotoxicity, in Fischer rats, but did not in WI rats. This difference in hepatotoxic response to Cd was supported by pathological analysis. After treatment with Cd at doses of 3.0, 3.5 and 4.5 mg/kg, the hepatic and renal accumulation of Cd was significantly lower in the WI rats than in the Fischer rats, indicating a kinetic mechanism for the observed strain differences in Cd toxicity. Thus, the remarkable resistance to Cd-induced hepatotoxicity in WI rats is associated, at least in part, with a lower tissue accumulation of the metal. Hepatic and renal zinc (Zn) contents after administration were similarly lower in WI than in Fischer rats. When Zn was administered in combination with Cd to Fischer rats, it decreased Cd contents in the liver and kidney, and exhibited a significant protective effect against the toxicity of Cd. We propose the possibility that Zn transporter plays an important role in the strain difference of Cd toxicity in WI and Fischer rats. PMID- 15363595 TI - Iron-induced interleukin-6 gene expression: possible mediation through the extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways. AB - Increased iron store in the body may increase the risk of many diseases such as cancer and inflammation. However, the precise pathogenic mechanism of iron has not yet been elucidated. In the present study, the early biological responses of cells to iron treatment were investigated in AP-1 luciferase reporter stably transfected mouse epidermal JB6 cells and primary rat hepatocytes. It was shown that water-soluble iron compounds, such as FeSO4 and Fe2(SO4)3, were more active in inducing AP-1 in JB6 cells than water-insoluble iron compounds, such as Fe2O3 and FeS. Iron stimulated mitogen-activated protein kinase (MAPK) family members of extracellular signal-regulated kinases (ERKs) and p38 MAPK but not c-jun NH2 terminal kinases (JNKs), both in JB6 cells and in primary rat hepatocytes, as determined by the phosphorylation assay. Interestingly, the increase in AP-1 luciferase activity by iron was inhibited by the pretreatment of the cells with PD98059, a specific MEK1 inhibitor, and SB202190, a p38 kinase inhibitor. Levels of interleukin-6 (IL-6), a pro-inflammatory cytokine, were increased in JB6 cells by iron in a dose-dependent manner. The increase in IL-6 and its mRNA by iron was also eliminated by the pretreatment of the cells with PD98059 and SB202190. Since the IL-6 promoter contains an AP-1 binding site, our studies indicate that the iron-induced IL-6 gene expression may be mediated through ERKs and p38 MAPK pathways, possibly one of the important mechanisms for the pathogenesis of iron overload. PMID- 15363596 TI - Effects of the phytoestrogen coumestrol on RANK-ligand-induced differentiation of osteoclasts. AB - Phytoestrogens, which have structural similarity to 17beta-estradiol, have been reported to act as agonists/antagonists of estrogen in animals and humans. Estrogen is known to have an important role in maintaining bone mass, because the concentration of serum estrogen decreases after menopause and the estrogen deficiency causes bone loss. In this study, we investigated the effects of coumestrol and other phytoestrogens on osteoclast differentiation using estrogen receptor alpha-transfected RAW264.7 (RAW264.7-ERalpha) cells. When the cells were cultured with the receptor activator of nuclear factor kappa B-ligand (RANKL), both formation of tartrate-resistant acid phosphatase (TRAP) positive multinucleated cells and TRAP activity were increased compared with control cells that were cultured in the absence of RANKL. Coumestrol decreased RANKL-induced formation of TRAP-positive multinucleated cells and TRAP activity dose dependently. RANKL-stimulated RAW264.7-ERalpha cells formed resorption pits on calcium phosphate films and the pit formation was inhibited by coumestrol in a dose-dependent manner. RT-PCR analyses revealed that coumestrol (10 microM) decreased mRNA levels of calcitonin receptor (CTR) and matrix metalloproteinase-9 (MMP9) in RANKL-treated cells. In addition, pretreatment of coumestrol decreased RANKL-induced phosphorylation of extracellular signal-regulated kinases/p44/42 (ERK1/2). These results suggest that coumestrol has an inhibitory effect on the differentiation of osteoclasts, at least partially via ERK1/2 pathway. PMID- 15363597 TI - Diverse developmental toxicity of di-n-butyl phthalate in both sexes of rat offspring after maternal exposure during the period from late gestation through lactation. AB - To evaluate developmental toxicity of di-n-butyl phthalate (DBP) with exposure during the period from late gestation to following lactation, maternal rats were given DBP at dietary concentrations of 0, 20, 200, 2000 and 10,000 ppm from gestational day 15 to postnatal day (PND) 21. At 10,000 ppm, male offspring showed a decreased neonatal anogenital distance and retention of nipples (PND 14), while females showed a slight non-significant delay in the onset of puberty. At PND 21, reduction of testicular spermatocyte development was evident from 20 ppm, as well as mammary gland changes at low incidence in both sexes. At this time point, population changes of pituitary hormone-immunoreactive cells were observed at 10,000 ppm with a similar pattern of increase in the percentages of luteinizing hormone (LH)-positive and decrease in follicle-stimulating hormone (FSH) and prolactin producing cells in both sexes, effects also being evident on FSH from 200 ppm and LH from 2000 ppm in females. During postnatal week (PNW) 8 11, marginal increase of the number of cases with extended diestrus was found at 10,000 ppm. At adult stage necropsy, testicular lesions appeared to be very faint in most cases, but degeneration and atrophy of mammary gland alveoli were observed in males from 20 ppm. Although without clear monotonic dose-dependence, relative pituitary weights were increased with the intermediate doses in males at PNW 11. In females, relative pituitary weights were decreased after 10,000 ppm at PNW 11, and from 200 ppm at PNW 20. The proportion of FSH-positive cells in the pituitaries at PNW 11 was increased in both sexes at 10,000 ppm. Thus, developmental exposure to DBP affected female sexual development involving pituitary function, while in males testicular toxicity was mostly reversible but mammary gland toxicity was persistent at a dose level as low as 20 ppm. PMID- 15363598 TI - Augmented fumonisin B1 toxicity in co-cultures: evidence for crosstalk between macrophages and non-parenchymatous liver epithelial cells involving proinflammatory cytokines. AB - Fumonisin B1, a common mycotoxin produced by Fusarium verticillioides found in corn, causes several fatal animal diseases. Liver and kidney are target organs of fumonisin B1 in laboratory animals, but primary rodent hepatocytes and liver slices were resistant to fumonisin B1-induced cytotoxic effects. We have shown that fumonisin B1 induces expression of tumor necrosis factor (TNF)alpha, interferon (IFN)gamma, and interleukine (IL) 12, in mouse liver. In various models of acute liver injury, a positive amplification loop involving TNFalpha, IFNgamma, and IL-12 has been implied that involves Kupffer cells (macrophages), hepatic lymphocytes and non-parenchymatous liver epithelial cells (NPECs). In the current study, cellular interactions in fumonisin B1-induced toxicity were investigated, using co-cultures of murine macrophages (J774A.1) and NPECs (NMuLi). Treatment of the co-cultures with fumonisin B1-produced cytotoxicity, whereas either J774A.1 or NMuLi cultures alone showed no response to the mycotoxin. Accumulation of sphinganine occurred to the similar extent in individual cultures as well as co-cultures. Expression of TNFalpha and IL-12 was increased in co-cultures but not in individual cultures. Transfer of conditioned medium from fumonisin B1-treated J774A.1 cells to NMuLi cultures produced an increase in IFNgamma expression in NMuLi cells. Results indicated that macrophages and liver epithelial cells interact in response to fumonisin B1 and potentiate the cytokines expression, which may have implications in making hepatocytes responsive to cytotoxicity of fumonisin B1. PMID- 15363599 TI - Avant-propos. PMID- 15363601 TI - Neuropsychopharmacology and the genetics of schizophrenia: a history of the diagnosis of schizophrenia. AB - Development of the diagnostic concept of schizophrenia (dementia praecox) is traced from the fourth edition of [Kraepelin, E., 1893. Ein Kurzes Lehrbuch der Psychiatrie. 4 Aufl. Barth, Lepzig] textbook to the DSM-IV [American Psychiatric Association, 1994. Diagnostic and Statistical Manual of Mental Disorders. Fourth ed. American Psychiatric Association, Washington, 273-316]. The differences between the criteria used by Bleuler [Bleuler, E., 1911. Dementia Praecox oder Gruppe der Schizophrenien. Deuticke, Leipzig] and Schneider [Fortschr. Neurol. Psychiatr. 25 (1957) 487] in the diagnosis of schizophrenia are discussed. The nosologic contributions of Kleist [Klin. Wochenschr. 2 (1923) 962] and Leonhard [Leonhard, K., 1957. Aufteilung der endogenen Psychosen. Akademie, Berlin]--which split schizophrenia into two major classes of disease with several forms and subforms--are outlined. Epidemiological findings--which show wide variations in the prevalence of schizophrenia in the general population and in the admission rate of schizophrenics to psychiatric clinics--are presented. Findings in genetic studies are reviewed with special reference to family, twin, and adoption studies which have raised the possibility that heredity plays an important role in the etiology of schizophrenia; mathematical analyses are examined which have ruled out monogenic transmission, as well as molecular genetic investigations indicating that the schizophrenic population is genetically heterogeneous. The relevance of findings with endophenotypes to the genetics of schizophrenia is questioned. The history of pharmacotherapy, neuropharmacology, and psychopharmacology of schizophrenia is outlined. Attention is focused on findings which indicate that the schizophrenic population is pharmacologically heterogenous. It is emphasized that neuropsychopharmacology, through its unique capability of linking the effects of psychotropic drugs to brain structures- encoded by genes which have been identified--offers a pioneering methodology for bridging the gap between the genes and psychiatric nosology. It is pointed out that for the detection of subpopulations within schizophrenia, clinical investigations with antipsychotic drugs have to proceed beyond the demonstration of therapeutic efficacy to the identification of treatment-responsive form(s) of illness. Early findings by Fish [L'Encephale 53 (1964) 245] are presented which indicate that affect-laden paraphrenia, one of the three forms of unsystematic schizophrenia in Leonhard [Leonhard, K., 1957. Aufteilung der endogenen Psychosen. Akademie, Berlin] classification, is the treatment-responsive subpopulation within schizophrenia for typical antipsychotic drugs. It is suggested that if the findings of Fish [L'Encephale 53 (1964) 245] could be verified, affect-laden paraphrenia would qualify for molecular genetic research. Another possible subpopulation that might qualify for genetic research is systematic hebephrenia, one of the three forms of systematic hebephrenia. The paper concludes that resolving the heterogeneity of the schizophrenic population would open up a new perspective for genetic research and for the pharmacotherapy of the different illnesses covered up for a century by the diagnostic label of schizophrenia. PMID- 15363602 TI - Platelets provide human tissue to unravel pathogenic mechanisms of Alzheimer disease. AB - Alzheimer disease (AD) is a progressive neurodegenerative disorder characterised by a progressive cognitive and memory decline. From a neuropathological point of view, Alzheimer disease is defined by the presence of characteristic lesions, i.e. mature senile plaques, neurofibrillary tangles and amyloid angiopathy. In particular, accumulation of the amyloid beta-peptide in the brain parenchyma and vasculature is an invariant event in the pathogenesis of both sporadic and familial Alzheimer cases. Amyloid beta-peptide originates from a larger precursor, the Amyloid Precursor Protein (APP) ubiquitously expressed. Among the different peripheral cells expressing APP forms, platelets are particularly interesting since they show concentrations of its isoforms equivalent to those found in brain. Moreover, a number of laboratories independently described alterations in APP metabolism/concentration in platelets of Alzheimer patients when compared to control subjects matched for demographic characteristics. These observations defined the frame of our work aimed to investigate if a correlation between levels of platelet APP forms and Alzheimer disease could be detected. We have reported that patients affected by Alzheimer disease show a differential level of platelet APP forms. This observation has several implications: APP processing abnormalities, believed to be a very early change in Alzheimer disease in neuronal compartment, does occur in extraneuronal tissues, such as platelets, thus suggesting that Alzheimer disease is a systemic disorder; further, our data strongly indicate that a differential level of platelet APP forms can be considered a potential peripheral marker of Alzheimer disease allowing for discrimination between Alzheimer and other types of dementia with good sensitivity and specificity. PMID- 15363603 TI - Role of oxidative stress in neurodegeneration: recent developments in assay methods for oxidative stress and nutraceutical antioxidants. AB - Reactive oxygen species (ROS) are produced in the course of normal metabolism and they serve important physiological functions. However, because of their high reactivity, accumulation of ROS beyond the immediate needs of the cell may affect cellular structure and functional integrity, by bringing about oxidative degradation of critical molecules, such as the DNA, proteins, and lipids. Although cells possess an intricate network of defense mechanisms to neutralize excess ROS and reduce oxidative stress, some tissues, especially the brain, are much more vulnerable to oxidative stress because of their elevated consumption of oxygen and the consequent generation of large amounts of ROS. For the same reason, the mitochondrial DNA (mtDNA) of brain cells is highly susceptible to structural alterations resulting in mitochondrial dysfunction. Several lines of evidence strongly suggest that these effects of ROS may be etiologically related to a number of neurodegenerative disorders. Nutraceutical antioxidants are dietary supplements that can exert positive pharmacological effects on specific human diseases by neutralizing the negative effects of ROS. The present communication concentrates on a review of recent concepts and methodological developments, some of them based on the results of work from our own laboratory, on the following aspects: (1) the complex interactions and complementary interrelationships between oxidative stress, mitochondrial dysfunction, and various forms of neural degeneration; (2) fractionation and isolation of substances with antioxidant properties from plant materials, which are extensively used in the human diet and, therefore, can be expected to be less toxic in any pharmacological intervention; (3) recent developments in methodologies that can be used for the assay of oxidative stress and determination of biological activities of exogenous and endogenous antioxidants; and (4) presentation of simple procedures based on polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of the resulting amplicon for investigations of structural alterations in mtDNA. PMID- 15363604 TI - N-methyl-D-aspartate antagonists as drug models of schizophrenia: a surprising link to tobacco smoking. AB - The pros and cons of N-methyl-D-aspartate (NMDA) antagonists as drug models of schizophrenia are discussed in relation to the neuropathology of this complex mental spectrum of diseases. The role of acetylcholine, dopamine, gamma aminobutyric acid, glutamic acid, and serotonin emphasizes that multiple neurotransmitter system abnormalities are involved, even though current drug therapy involves primarily dopamine (D(2))/serotonin (5 HT(2)) antagonists. Only some of the fundamental aspects of schizophrenia are replicated by NMDA receptor antagonists of glutamic acid. Subchronic NMDA antagonism in an animal model results in decreased levels of dopamine in prefrontal cortex and increased D(1) receptor binding. The results of PET studies of schizophrenic patients imply decreased dopamine levels in their prefrontal cortex. Tobacco-smoking schizophrenic patients transiently normalize prepulse inhibition. Nicotine appears to be one factor that may help explain some of these phenomena. PMID- 15363605 TI - Ephrin signaling in axon guidance. AB - The multiple functions of a neuron depend on the proper assembly of axonal connections during the development of the nervous systems. This assembly involves the motile behavior of growth cones at the ends of elongating axons. The growth cones express receptors that bind to specific guidance molecules in the local environment. In turn, this initiates the attractive and repulsive forces required to give the appropriate direction to the elongating axon. The process implicates a tightly regulated remodeling of the actin cytoskeleton in response to the activation of the Rho GTPases, Cdc42, Rac and RhoA. In this article, we will review how the ephrin-Eph receptor system regulates the activity of the Rho GTPases, to modulate the mechanics of growth cone activity and then axon guidance. PMID- 15363606 TI - Role of presynaptic alpha2-adrenoceptors in antidepressant action: recent findings from microdialysis studies. AB - The therapeutic effect of an antidepressant drug takes at least 2 to 3 weeks to develop and a significant proportion of patients have no or only partial benefit regardless of the class of antidepressant used. Research into the neurobiological basis of antidepressant action has suggested new strategies to improve the antidepressant effect. Recent microdialysis studies show that hypofunction of the presynaptic autoreceptors enhances the increase of extracellular serotonin (5-HT) induced by selective serotonin reuptake inhibitors (SSRIs) so it has been suggested that the antidepressant effect may be speeded up by blockade of the autoreceptors. The similarity between the synaptic mechanisms controlling serotonergic and noradrenergic transmission has stimulated preclinical research into the role of presynaptic alpha(2)-adrenoceptors in the effect of noradrenaline (NA) reuptake inhibitors (NRIs) on NA availability at central synapses. The microdialysis studies reviewed here indicate that NRIs including desipramine, reboxetine and atomoxetine, the mixed 5-HT/NA reuptake inhibitors sibutramine, duloxetine, venlafaxine or the NA/DA reuptake inhibitor amineptine, increased extracellular NA in various regions of the rat brain. The effect was enhanced by chronic treatment and even more by the co-administration of alpha(2) adrenoceptor antagonists. The results support the theory that desensitization of the alpha(2)-adrenoceptor contributes to enhancing the effect of NRIs seen after chronic administration and may account for the slow onset of the antidepressant effect. Finally, they suggest that co-administration of an alpha(2)-adrenoceptor antagonist may improve the therapeutic effect of NRI. PMID- 15363607 TI - Noradrenaline storage function of species-specific protein bodies, markers of monoamine neurons in human locus coeruleus demonstrated by dopamine-beta hydroxylase immunogold localization. AB - Our histochemical and ultrastructural studies have identified, in human catecholamine locus coeruleus (LC) neurons, abundant and large spherical protein bodies (PB), containing histone-like, arginine-rich proteins, which originate as dense bodies in mitochondria. This species-specific phenotype in the neurons of man is highly intriguing. In the electron microscope PB are disrupted in LC neurons in depressed individuals, where noradrenaline is known to be reduced. This coincidence of ultrastructure and neurochemistry raises the question whether these bodies could qualify as noradrenaline-storing organelles in the human LC. Our rationale was to examine, in known model tissues that contain catecholamines- sympathetic ganglia and tumors of the autonomic nervous system--if vesicles show the same fine structure and histochemistry as the PB of the human LC. Hence, we selected biopsy tissues of five ganglioneuromas and postmortem tissues of LC from 25 control subjects. Since dopamine-beta-hydroxylase (DBH) is a hallmark of noradrenaline identity and present in dense core vesicles, the investigation of DBH localization with the immunogold method constituted the experiment of choice for this study. Histochemical determinations of arginine with Carmoisine L, and of lipids with Rhodamine B complemented the study of similarities between the PB of the human LC and ganglioneuromas. Our results showed, with the colloidal gold method, that DBH immunogold labeling was localized in the core and in the double membranes of the PB, and also in the adjacent mitochondria. These results indicate that protein bodies (a) are unequivocal storage vesicles of noradrenaline, and (b) derive from regular mitochondria and represent a new phenotype in man, which is probably an evolutionary adaptation of amine-storing organelles. PMID- 15363608 TI - Adverse effects of cannabis on health: an update of the literature since 1996. AB - Recent research has clarified a number of important questions concerning adverse effects of cannabis on health. A causal role of acute cannabis intoxication in motor vehicle and other accidents has now been shown by the presence of measurable levels of Delta(9)-tetrahydrocannabinol (THC) in the blood of injured drivers in the absence of alcohol or other drugs, by surveys of driving under the influence of cannabis, and by significantly higher accident culpability risk of drivers using cannabis. Chronic inflammatory and precancerous changes in the airways have been demonstrated in cannabis smokers, and the most recent case control study shows an increased risk of airways cancer that is proportional to the amount of cannabis use. Several different studies indicate that the epidemiological link between cannabis use and schizophrenia probably represents a causal role of cannabis in precipitating the onset or relapse of schizophrenia. A weaker but significant link between cannabis and depression has been found in various cohort studies, but the nature of the link is not yet clear. A large body of evidence now demonstrates that cannabis dependence, both behavioral and physical, does occur in about 7-10% of regular users, and that early onset of use, and especially of weekly or daily use, is a strong predictor of future dependence. Cognitive impairments of various types are readily demonstrable during acute cannabis intoxication, but there is no suitable evidence yet available to permit a decision as to whether long-lasting or permanent functional losses can result from chronic heavy use in adults. However, a small but growing body of evidence indicates subtle but apparently permanent effects on memory, information processing, and executive functions, in the offspring of women who used cannabis during pregnancy. In total, the evidence indicates that regular heavy use of cannabis carries significant risks for the individual user and for the health care system. PMID- 15363609 TI - Molecular genetics of affective disorders. AB - Evidence for familial aggregation in Affective Disorders (AD) has been provided in classical studies. Linkage and association genetic studies have been proposed to detect genetic factors implicated in AD. However, findings from molecular genetic studies remain inconclusive. Nevertheless, current research is focusing on the phenotypes, both sub- and endophenotypes. In addition, recent advances in technology, such as microarrays, provide new tools in psychiatric genetics. These different approaches offer a new optimism era in the search of genetic factors in AD. PMID- 15363610 TI - Pilot study on the use of psychotropic medication in persons with mental retardation. AB - OBJECTIVE: To explore the status of the psychotropic medication in persons with mental retardation 6 years after their deinstitutionalization. METHOD: The authors compared the profiles of psychotropic medication of 60 persons with mental retardation deinstitutionalized (D-sample) from our local mental hospital 6 years ago, to their actual pharmacological profile. Later, we compared our results to the ones of persons with mental retardation on an in-patient (In sample) unit where existed a concerted team effort to lower the medication when deemed appropriate. RESULTS: Six years post-deinstitutionalization, the authors observed that the same percentage of patients remained on antipsychotic and benzodiazepine medication in our D-sample, and a threefold increase of those using antidepressive medication. However, we observe a general decrease of all types of psychotropic medication in the In-sample with complete withdrawal in a substantial number of patients. CONCLUSIONS: The authors take these pilot data to mean that a concerted team effort at lowering medication in persons with mental retardation makes a significant difference. The possibility of other factors (age, sex, severity of mental retardation, psychiatric diagnosis) explaining the difference in the results is discussed. PMID- 15363611 TI - Social and medical origins of neurochemistry. AB - Neurochemistry, the biochemistry of brain and nerve, had its identifiable scientific origins in France just before the French Revolution. Its main development remained in the hands of Parisian chemists over the next four decades. Thereafter, interest in the subject spread slowly through the German speaking world. The most significant advances were made by a Hessian, practicing medicine and chemistry as an emigre in London in the second half of the 19th century. Some chemists had an intrinsic interest in exploring the brain by the techniques which they had applied to various organs of the body. However, there were others for whom social, medical, even philosophical, factors were the spur in their choice of study of the chemistry of the nervous system. This paper describes the early growth of neurochemistry as I have come to understand it during the course of historical research during the past 10 years. PMID- 15363612 TI - Can stress cause depression? AB - The central issue raised in this paper is: can stress cause depression? Phrased more precisely: can stress cause brain disturbances thought to underlie (certain forms of) depression or particular components of the depressive syndrome. Focussing on 5-hydroxytryptamine (5-HT) and the stress hormones, this question was answered in the affirmative, based on the following two considerations: changes in the 5-HT and stress hormone systems produced by sustained stress mimic to a substantial extent the disturbances in these systems that may be observed in depression. Substantial evidence indicates that the 5-HT and stress hormone disturbances in depression are of pathophysiological significance and not merely a consequence of the depressed state or a product of stress generated by the depressed state. Furthermore, the question was raised whether a depression type could be identified particularly stress-inducible. This question, too, was answered in the affirmative. The depression type in question was named anxiety/aggression-driven depression and characterized on three levels: psychopathologically, biologically and psychologically. Preferential treatment of this depression type was discussed. In studying stress-inducible depression, biological depression research should shift focus from depression per se to the neurobiological sequelae of stress. Treatment of stress-inducible depressions and particularly its prevention should be geared towards reduction of stress and stress sensitiveness, utilising both biological and psychological means. PMID- 15363613 TI - Neurobiological differences on populational level. AB - At present, 13% of world population are suffering from neuropsychiatric disorders. Prevention and treatment of these pathological conditions are very important. New research strategies are needed for the creation of effective methods for controlling of neuropsychiatric disorders. International collaborative research projects within WHO's activities in psychopharmacology and biological psychiatry with the participation of 12 countries representing different world regions have been organized and successfully finalized. The author discusses some of these in the Festschrift for Dr. Corneille Radouco Thomas. PMID- 15363614 TI - Proposal for a noradrenaline hypothesis of schizophrenia. AB - In this article, we have reevaluated the role of noradrenergic dysfunction in the pathogenesis of schizophrenia in the light of today's neuroscience and clinical data. Neurophysiological, psychophysiological, psychopharmacological, and biochemical findings that have accumulated in last decades indicate that certain noradrenergic dysfunctions play important roles in the pathogenesis of the disorder. Moreover, these findings provide us with consistent evidence for the existence of two syndromes generated by either overactivity or underactivity of the central noradrenaline (NA) system. The former appears to correspond to the type I syndrome (positive symptoms) and the latter to the type II syndrome (negative symptoms). We conclude that the involvement of brain NA in cerebral metabolism and blood flow as well as the amine's role in brain development and neuronal differentiation may provide the mechanisms underlying the disease process in schizophrenia. Development of chemical agents acting specifically on the brain noradrenergic mechanisms may be a promising approach to novel treatments of the disorder. PMID- 15363615 TI - [How useful is the videocapsule endoscopy in 2004?]. PMID- 15363616 TI - [Anti-Ku antibodies. Study of prevalence and of clinical meaning]. AB - PURPOSE: Auto-immunity against Ku nuclear antigens is rare and clinical meaning remains badly estimated. Our study is for purposes: to appreciate the prevalence of antibodies anti-Ku within the framework of the search for antinuclear antibodies and to clarify clinical and biological relations associated to this auto-immunity. METHODS: A retrospective study of a series of 10,000 searches for antinuclear antibodies studies the prevalence of the auto-immunity anti-Ku and a retrospective analysis of the data found at the patients bearers of an anti-Ku identifies clinical and biological signs associated with this antibody. RESULTS: Prevalence anti-Ku is low (1/3493 case of antinuclear antibodies) and association is possible with in a myositic process through variable auto-immune contexts (overlap syndrome) of relative good preview. CONCLUSION: Auto-immunity anti-Ku is so characterized with its weak prevalence, a possible observation during different auto-immune diseases with an obvious frequency of the overlap syndrome often concerning a process myositic. Finally a weak evolutivite seems to characterize the auto-immune diseases of the patients with anti-Ku antibodies. PMID- 15363617 TI - [Haemochromatosis screening in 120 patients complaining with persistant fatigue]. AB - AIM: Chronic fatigue is the more frequent symptom identified in the course of hereditary haemochromatosis. A screening for this disorder was carried out in 120 primary care patients consulting for unexplained chronic fatigue. SUBJECTS AND METHODS: Transferrin saturation and serum ferritin were determined in all patients. If transferrin saturation was >or= 45% and serum ferritin >or= 300 microg/l, HFE1 genotyping for mutations C282Y and H63D was completed. RESULTS: One hundred and twenty patients were recruited, 19-86 years old, including 62 males and 58 females. 45 patients (38%) presented with serum ferritin >or= 300 microg/l. Thirty two patients (27%) presented with transferrin saturation >or= 45%. Twenty two patients (18%) presented with these two pathological values. Four C282Y/H63D compound heterozygous, one H63D/H63D homozygous, and eight simplex heterozygous (6 H63D and 2 C282Y) genotypes were found. Patients with serum ferritin >or= 300 microg/l were predominantly male (89%), older (57 year) and plethoric (BMI: 26.4) corresponding mainly to dysmetabolic hyperferritinemia. CONCLUSION: None of these 120 patients consulting for unexplained chronic fatigue was found with hereditary haemochromatosis. Therefore observed prevalence is 0, with upper limit of 95% confidence interval at 2.5%. But the high prevalence (38%) of serum ferritin >or= 300 microg/l must be emphasized, corresponding usually to dysmetabolic hyperferritinemia. PMID- 15363618 TI - [Cationic anti-microbial peptides: from innate immunity study to drug development]. AB - OBJECTIVE: Host defense, cationic anti-microbial peptides are recognized as an important component of innate immune response in most multicellular organisms. NEW FEATURES: They are cationic amphipathic peptides, comprising 20-50 amino acids. Several hundreds of peptides have been characterized. They have broad spectrum of activity against bacterial, fungal and viral pathogens. The mode of action is best known for cecropins and magainins, which act upon the cytoplasmic membrane of microorganisms, causing its disruption by a detergent like activity and pore formation. Several of these peptides or analogs (from magainin, protegrin, indolicidin and histatin) are in advanced clinical development, especially for localized infections. PROJECTS AND PERSPECTIVES: Several other molecules (rBPI, heliomicin and thanatine) are currently under development for various systemic infections. They may represent important drugs of the further anti-infectious therapeutic arsenal. PMID- 15363619 TI - [Glucocorticoids and their receptor: mechanisms of action and clinical implications]. AB - BACKGROUND: Glucocorticoids are used as anti-inflammatory, immuno-modulatory, anti-proliferative and cytotoxic drugs, but they also trigger important side effects. These hormones bind to glucocorticoid receptor alpha (GRalpha), an intracellular protein, which acts essentially in the nucleus. MAIN POINTS: GRalpha is a ligand-activated transcription factor that positively or negatively regulates gene expression by distinct mechanisms. Stimulation of gene transcription occurs after direct binding of the receptor to specific responsive DNA elements. Gene activation by glucocorticoids is mainly responsible for certain adverse effects. In contrast, the therapeutic effects of glucocorticoids are predominantly mediated through repression of genes encoding inflammatory mediators. Inhibitory protein-protein interaction between the hormone-activated receptor and the transcription factors NF-kappaB and AP-1 was found to be the underlying mechanism. However, inhibition of other transcription factors may account for deleterious effects of glucocorticoids, such as adrenal suppression and osteoporosis. GRalpha also mediates rapid non-genomic effects of glucocorticoids. Side-effects are reduced by using topical glucocorticoids which have a low systemic bioavailability. Moreover, it is important to determine the lowest effective maintenance dose of systemic and topical glucocorticoids to further decrease the risk of adverse effects. This is particularly justified because inhibition of AP-1 and NF-kappaB activities, that is the anti inflammatory effect, occurs at much lower hormone concentrations than transactivation. PERSPECTIVES: Clinical use of glucocorticoids is limited by occurrence of severe adverse effects. Therefore, the current aim is to design GRalpha ligands that retain only the anti-inflammatory activities of GC. PMID- 15363620 TI - [Pathogenic mechanisms of autoimmune diseases]. AB - BACKGROUND: Auto-immune diseases are a fascinating but poorly understood group of diseases. MAJOR POINTS: In this review, we will consider genetic susceptibility to auto-immune diseases, initiation of autoreactivity and changes in pathologic processes. Recent physiopathological hypotheses are detailed with clinical examples. PERSPECTIVES: A better knowledge of the physiopathological mechanisms would allow the development of more efficacious treatments. PMID- 15363621 TI - [Violacein: a molecule of biological interest originating from the soil-borne bacterium Chromobacterium violaceum]. AB - INTRODUCTION: Soil micro-organisms have evolved functions that allow them to withstand the strong competition for survival that characterizes most of their habitats. The production of antibiotic or antifungal compounds is one of these mechanisms. The relevant molecules often exhibit valuable therapeutic properties. EXEGESIS: Chromobacterium violaceum is a soil-borne bacterium producing a characteristic antibiotic termed violacein. It is part of a series of compounds released by C. violaceum to oppose competitors and predators in the soil and water environments. Violacein, and one of these compounds, i.e. structure FR901228, exhibit antiparasitic and antitumoral activities of potential medical interest. Genes involved in the synthesis of these compounds are available, the genome sequence of C. violaceum (strain ATCC 12472) being published. CONCLUSIONS: The above example, involving Chromobacterium, is not an exception: soil constitutes a reservoir of molecules, enzymatic activities and micro-organisms of biological interest, the study of which will undoubtedly lead to developments in fields as diverse as agronomy or animal and human therapeutics. PMID- 15363622 TI - [Icteric Fort Bragg fever: a case report with nosological discussion]. AB - INTRODUCTION: Ictero-hemorrhagic leptospirosis is an endemic disease in France. Weil's disease, a form of leptospirosis, is well known. Fort Bragg fever is characterized by a constant pretibial papular lesion. First described in the USA, this non icteric form of leptospirosis is usually benign. We report the first French case of a mixed form of leptospirosis. EXEGESIS: A 52-year-old man living in South East France suffered from fever and myalgias associated with a pretibial papular lesion. A severe icterus appeared and permitted a diagnosis of leptospirosis. CONCLUSION: Our case recalls the clinical presentation of Fort Bragg fever, which is recognized through its inflammatory pretibial lesion associated or not with icterus. PMID- 15363623 TI - [Acquired C1 inhibitor deficiency associated with lymphoproliferative disorders: four cases]. AB - INTRODUCTION: Acquired C1 inhibitor deficiency is sometimes associated with lymphoproliferative disorders. EXEGESIS: We report four cases of acquired C1 inhibitor deficiency in association with lymphoproliferative disorders. Three of them were asymptomatic; one was associated with abdominal pain. Four women (median age, 66 years) presented either two non-Hodgkin lymphoma or two chronic lymphocytic leukaemia. C1 inhibitor deficiency was detected fortuitous (n = 1) or during investigation of arthralgia (n = 2), or Gougerot-Sjogren syndrome (n = 1). The deficit was acquired in all cases type I. Auto-immune disorders were associated with: Gougerot-Sjogren syndrome (n = 1), cryoglobulinemia (n = 2), IgM lambda monoclonal gammopathy (n = 1), Coombs positive test (n = 2), IgG anti cardiolipine antibodies (n = 1). C1 inhibitor deficiency was not modified after lymphoproliferative disorders treatment (radiotherapy, splenic ablation) in two cases but patients were not in complete remission. C1 inhibitor raised normal level in one case, after five chemotherapy regimens, but decreased complement level and C4 split persist. CONCLUSION: Acquired C1 inhibitor deficiency associated with lymphoproliferative disorders is sometimes asymptomatic. Diagnosis could be delay in spite of clinical manifestations. Deficit correction is not constant after lymphoproliferative disorders treatment. PMID- 15363624 TI - [Gastrointestinal bleeding due to a gastric stromal tumor]. PMID- 15363625 TI - [Amyloidosis in adult onset Still's disease]. PMID- 15363626 TI - [Rheumatoid arthritis and Kikuchi's lymphadenitis: a case-report and review of the literature]. PMID- 15363627 TI - ["Actinobacillus and Haemophilus parainfluenzae infective endocarditis: two case reports"]. PMID- 15363628 TI - [Fibrous solitary tumour of neck]. PMID- 15363629 TI - Elucidation of the ryanodine-sensitive Ca2+ release mechanism of rat pancreatic acinar cells: modulation by cyclic ADP-ribose and FK506. AB - The effects of cyclic ADP-ribose (cADPR) and the immunosuppressant drug FK506 on microsomal Ca2+ release through a ryanodine-sensitive mechanism were investigated in rat pancreatic acinar cells. After a steady state of 45Ca2+ uptake into the microsomal vesicles, ryanodine or caffeine was added. Preincubation of the vesicles with cADPR (0.5 microM) shifted the dose-response curve of ryanodine- or caffeine-induced 45Ca2+ release from the vesicles to the left. Preincubation with cADPR shifted the dose-response curve of the FK506-induced 45Ca2+ release upward. Preincubation with FK506 (3 microM) shifted the dose-response curve of the ryanodine- or caffeine-induced 45Ca2+ release to the left by the same extent as that in the case of cADPR. FK506 shifted the dose-response curve of the cADPR induced 45Ca2+ release upward. The presence of both cADPR and FK506 enhanced the ryanodine (30 microM)- or caffeine (10 mM)-induced 45Ca2+ release by the same extent as that in the case of cADPR alone or FK506 alone. These results indicate that cADPR and FK506 modulate the ryanodine-sensitive Ca2+ release mechanism of rat pancreatic acinar cells by increasing the ryanodine or caffeine sensitivity to the mechanism. In addition, there is a possibility that the mechanisms of modulation by cADPR and FK506 are the same. PMID- 15363630 TI - Gene expression profiling after treatment with the histone deacetylase inhibitor trichostatin A reveals altered expression of both pro- and anti-apoptotic genes in pancreatic adenocarcinoma cells. AB - The histone deacetylase inhibitor trichostatin A (TSA) has been previously shown to block cellular growth in G2 and induce apoptosis in human pancreatic cancer cell lines. In order to better understand this phenomenon, we have analyzed the gene expression profiles in PaCa44 cells after treatment with TSA using microarrays containing 22,283 probesets. TSA was found to cause both the induction and repression of a large number of genes, although the number whose expression was up-regulated was greater than the number of genes that were down regulated. When a threshold value of 3 was used as a cutoff level, a total of 306 (3.4%) of the detectable genes had altered expression. When categorized according to cellular function, the differentially expressed genes were found to be involved in a wide variety of cellular processes, including cell proliferation, signaling, regulation of transcription, and apoptosis. Moreover, Sp1/Sp3 transcription factor binding sites were significantly more abundant among TSA induced genes. One prominent feature was the increased ratio between the levels of expression of pro-apoptotic (BIM) and anti-apoptotic (Bcl-XL and Bcl-W) genes. This result was confirmed in eight additional pancreatic cancer cell lines after treatment with TSA, suggesting that this event may be a strong determinant for the induction of apoptosis by TSA. PMID- 15363631 TI - Calmodulin binds to and inhibits the activity of phosphoglycerate kinase. AB - Phosphoglycerate kinase (PGK) functions as a cytoplasmic ATP-generating glycolytic enzyme, a nuclear mediator in DNA replication and repair, a stimulator of Sendai virus transcription and an extracellular disulfide reductase in angiogenesis. Probing of a developmental expression library from Dictyostelium discoideum with radiolabelled calmodulin led to the isolation of a cDNA encoding a putative calmodulin-binding protein (DdPGK) with 68% sequence similarity to human PGK. Dictyostelium, rabbit and yeast PGKs bound to calmodulin-agarose in a calcium-dependent manner while DdPGK constructs lacking the calmodulin-binding domain (209KPFLAILGGAKVSDKIKLIE228) failed to bind. The calmodulin-binding domain shows 80% identity between diverse organisms and is situated beside the hinge and within the ATP binding domain adjacent to nine mutations associated with PGK deficiency. Calmodulin addition inhibits yeast PGK activity in vitro while the calmodulin antagonist W-7 abrogates this inhibition. Together, these data suggest that PGK activity may be negatively regulated by calcium and calmodulin signalling in eukaryotic cells. PMID- 15363632 TI - Combined effect of testosterone and apocynin on nitric oxide and superoxide production in PMA-differentiated THP-1 cells. AB - Human inducible nitric oxide synthase (iNOS) is most readily observed in macrophages from patients with inflammatory diseases like atherosclerosis. The aim of the present study was to find out the combined effect of male sex hormone; testosterone and apocynin (NADPH oxidase inhibitor) on cytokine-induced iNOS production. THP-1 cells were differentiated into macrophages by phorbol myristate acetate (PMA). Expression of iNOS was induced by the addition of cytokine mixture? Testosterone was added at different concentrations (10(-6)-10(-12) M) with apocynin (1 mM). Testosterone (10(-8), 10(-10) M) inhibited NOx production in cytokine-added THP-1 cells which was further confirmed by quantikine assay of iNOS protein and RT-PCR analysis. Testosterone treatment decreased 40% of superoxide anion production. Testosterone showed inhibition of NADPH oxidase, especially expression of p67phox and p47phox (cytosol subunits). Addition of testosterone with apocynin further decreased the expression of p67phox and p47phox subunits of NADPH oxidase. The findings of the present study suggest that, testosterone; the male androgen plays an important role in the prevention of atherogenesis. Even though apocynin does not have any role on NO production, addition of apocynin together with testosterone is effective in suppressing iNOS activity. PMID- 15363633 TI - Induction of tenascin-C by cyclic tensile strain versus growth factors: distinct contributions by Rho/ROCK and MAPK signaling pathways. AB - Expression of the extracellular matrix (ECM) protein tenascin-C is induced in fibroblasts by growth factors as well as by tensile strain. Mechanical stress can act on gene regulation directly, or indirectly via the paracrine release of soluble factors by the stimulated cells. To distinguish between these possibilities for tenascin-C, we asked whether cyclic tensile strain and soluble factors, respectively, induced its mRNA via related or separate mechanisms. When cyclic strain was applied to chick embryo fibroblasts cultured on silicone membranes, tenascin-C mRNA and protein levels were increased twofold within 6 h compared to the resting control. Medium conditioned by strained cells did not stimulate tenascin-C mRNA in resting cells. Tenascin-C mRNA in resting cells was increased by serum; however, cyclic strain still caused an additional induction. Likewise, the effect of TGF-beta1 or PDGF-BB was additive to that of cyclic strain, whereas IL-4 or H2O2 (a reactive oxygen species, ROS) did not change tenascin-C mRNA levels. Antagonists for distinct mitogen-activated protein kinases (MAPK) inhibited tenascin-C induction by TGF-beta1 and PDGF-BB, but not by cyclic strain. Conversely, a specific inhibitor of Rho-dependent kinase strongly attenuated the response of tenascin-C mRNA to cyclic strain, but had limited effect on induction by growth factors. The data suggest that regulation of tenascin-C in fibroblasts by cyclic strain occurs independently from soluble mediators and MAPK pathways; however, it requires Rho/ROCK signaling. PMID- 15363634 TI - Early insulin signaling cascade in a model of oxidative skeletal muscle: mouse Sol8 cell line. AB - Cell models provide important tools to investigate the mechanisms modulating the insulin-signaling cascade. Insulin interaction and subsequent signaling of cells is complex and regulated at multiple levels: receptor abundance, binding dynamics, phosphorylation/dephosphorylation of tyrosine and serine/threonine residues, and subsequent interactions of key intracellular messengers. We report early insulin signaling events in the mouse Sol8 myogenic cell line. Sol8 cells responded to insulin by increasing total IRS-1, p85 PI3-kinase and tyrosine phosphorylated IRS-1 (pY-IRS-1) at 10 min (P<0.05), but not at 1 min of insulin stimulation. The dose-response relationships at 10-min insulin (10 to 300 nM) stimulation showed that IRS-1 and pY-IRS-1 responded to 100 and 300 nM insulin, and the p85 PI3-kinase response peaked at 30 nM insulin. PI3-kinase appeared to be present in high abundance and, in response to insulin, recruitment to the insulin receptor tyrosine kinase (IR) of IRS-1 and PI3-kinase was observed. The increase in IRS-1 detected in IR immunoprecipitates was twofold, while the corresponding increase in PI3-kinase was threefold, suggesting direct recruitment of PI3-kinase to the IR. PI3-kinase detected in IRS-1 immunoprecipitates in response to insulin increased 1.7-fold. An ultimate target of this pathway, GLUT4 recruitment to the PM, was delayed (30 min), the increase in GLUT4 being of similar magnitude (1.6-fold) to the early signaling events. Saturation binding analysis indicated that IR in the plasma membrane was not down-regulated in response to insulin. The present study suggests that early signaling events in the insulin cascade are invoked in Sol8 myogenic cells and that this cell line provides a useful model to study insulin signaling. PMID- 15363636 TI - Carnitine: a nutritional, biosynthetic, and functional perspective. AB - Carnitine status in humans is reported to vary according to body composition, gender, and diet. Plasma carnitine concentration positively correlates with the dietary intake of carnitine. The content of carnitine in foodstuff is based on old and inadequate methodology. Nevertheless, dietary carnitine is important. The molecular biology of the enzymes of carnitine biosynthesis has recently been accomplished. Carnitine biosynthesis requires pathways in different tissues and is an efficient system. Overall biosynthesis is determined by the availability of trimethyllysine from tissue proteins. Carnitine deficiency resulting from a defect in biosynthesis has yet to be reported. The role of carnitine in long chain fatty acid oxidation is well defined. Recent evidence supports a role for the voltage-dependent anion channel in the transport of acyl-CoAs through the mitochondrial outer membrane. The mitochondrial outer membrane carnitine palmitoyltransferase-I in liver can be phosphorylated and when phosphorylated the sensitivity to malonyl-CoA is greatly decreased. This may explain the change in sensitivity of liver carnitine palmitoyltransferase-I observed during fasting and diabetes. Recently reported data clarify the role of carnitine and the carnitine transport system in the interplay between peroxisomes and mitochondrial fatty acid oxidation. Lastly, the buffering of the acyl-CoA/CoA coupled by carnitine reflects intracellular metabolism. This mass action effect underlies the use of carnitine as a therapeutic agent. In summary, these new observations help to further our understanding of the molecular aspects of carnitine in medicine. PMID- 15363637 TI - Carnitine acyltransferases and their influence on CoA pools in health and disease. AB - Cells contain limited and sequestered pools of Coenzyme A (CoA) that are essential for activating carboxylate metabolites. Some acyl-CoA esters have high metabolic and signalling impact, so control of CoA ester concentrations is important. This and transfer of the activated acyl moieties between cell compartments without wasting energy on futile cycles of hydrolysis and resynthesis is achieved through the carnitine system. The location, properties of and deficiencies in the carnitine acyltransferases are described in relation to their influence on the CoA pools in the cell and, hence, on metabolism. The protection of free CoA pools in disease states is achieved by excretion of acyl carnitine so that carnitine supplementation is required where unwanted acyl groups build up, such as in some inherited disorders of fatty acid oxidation. Acetyl-carnitine improves cognition in the brain and propionyl-carnitine improves cardiac performance in heart disease and diabetes. The therapeutic effects of carnitine and its esters are discussed in relation to the integrative influence of the carnitine system across CoA pools. Recent evidence for sequestered pools of activated acetate for synthesis of malonyl-CoA, for the synthesis of polyunsaturated fatty acids and for the inhibition of carnitine palmitoyltransferase 1 to regulate fatty acid oxidation is reviewed. PMID- 15363638 TI - Carnitine palmitoyltransferases 1 and 2: biochemical, molecular and medical aspects. AB - Carnitine palmitoyltransferase (CPT) deficiencies are common disorders of mitochondrial fatty acid oxidation. The CPT system is made up of two separate proteins located in the outer (CPT1) and inner (CPT2) mitochondrial membranes. While CPT2 is an ubiquitous protein, three tissue-specific CPT1 isoforms--the so called "liver" (CPT1-A), "muscle" (CPT1B) and <> (CPT1-C) CPT1s--have been shown to exist. Amino acid and cDNA nucleotide sequences have been identified for all of these proteins. CPT1-A deficiency presents as recurrent attacks of fasting hypoketotic hypoglycemia. Twenty four CPT1A mutations have been reported to date. CPT1-B and -C deficiencies have not been hitherto identified. CPT2 deficiency has several clinical presentations. The "benign" adult form (more than 200 families reported) is characterized by episodes of rhabdomyolysis triggered by prolonged exercise. The prevalent S113L mutation is found in about 50% of mutant alleles. The infantile-type CPT2 presents as severe attacks of hypoketotic hypoglycemia, occasionally associated with cardiac damage commonly responsible for sudden death before 1 year of age. In addition to these symptoms, features of brain and kidney dysorganogenesis are frequently seen in the neonatal-onset CPT2 deficiency, almost always lethal during the first month of life. Around 40 CPT2 mutations (private missense or truncating mutations) have hitherto been detected. Treatment is based upon avoidance of fasting and/or exercise, a low fat diet enriched with medium chain triglycerides and carnitine. Prenatal diagnosis may be offered for pregnancies at a 1/4 risk of infantile/severe-type CPT2 deficiency. PMID- 15363639 TI - Carnitine-acylcarnitine translocase deficiency, clinical, biochemical and genetic aspects. AB - The carnitine-acylcarnitine translocase (CACT) is one of the components of the carnitine cycle. The carnitine cycle is necessary to shuttle long-chain fatty acids from the cytosol into the intramitochondrial space where mitochondrial beta oxidation of fatty acids takes place. The oxidation of fatty acids yields acetyl coenzyme A (CoA) units, which may either be degraded to CO(2) and H(2)O in the citric acid cycle to produce ATP or converted into ketone bodies which occurs in liver and kidneys. Metabolic consequences of a defective CACT are hypoketotic hypoglycaemia under fasting conditions, hyperammonemia, elevated creatine kinase and transaminases, dicarboxylic aciduria, very low free carnitine and an abnormal acylcarnitine profile with marked elevation of the long-chain acylcarnitines. Clinical signs and symptoms in CACT deficient patients, are a combination of energy depletion and endogenous toxicity. The predominantly affected organs are brain, heart and skeletal muscle, and liver, leading to neurological abnormalities, cardiomyopathy and arrythmias, skeletal muscle damage and liver dysfunction. Most patients become symptomatic in the neonatal period with a rapidly progressive deterioration and a high mortality rate. However, presentations at a later age with a milder phenotype have also been reported. The therapeutic approach is the same as in other long-chain fatty acid disorders and includes intravenous glucose (+/- insulin) administration to maximally inhibit lipolysis and subsequent fatty acid oxidation during the acute deterioration, along with other measures such as ammonia detoxification, depending on the clinical features. Long-term strategy consists of avoidance of fasting with frequent meals and a special diet with restriction of long-chain fatty acids. Due to the extremely low free carnitine concentrations, carnitine supplementation is often needed. Acylcarnitine profiling in plasma is the assay of choice for the diagnosis at a metabolite level. However, since the acylcarnitine profile observed in CACT-deficient patients is identical to that in CPT2-deficient patients, definitive identification of CACT-deficiency in a certain patient requires determination of the activity of CACT. Subsequently, mutational analysis of the CACT gene can be performed. So far, 9 different mutations have been identified in the CACT gene. PMID- 15363640 TI - Role of carnitine esters in brain neuropathology. AB - L-Carnitine (L-C) is a naturally occurring quaternary ammonium compound endogenous in all mammalian species and is a vital cofactor for the mitochondrial oxidation of fatty acids. Fatty acids are utilized as an energy substrate in all tissues, and although glucose is the main energetic substrate in adult brain, fatty acids have also been shown to be utilized by brain as an energy substrate. L-C also participates in the control of the mitochondrial acyl-CoA/CoA ratio, peroxisomal oxidation of fatty acids, and the production of ketone bodies. Due to their intrinsic interaction with the bioenergetic processes, they play an important role in diseases associated with metabolic compromise, especially mitochondrial-related disorders. A deficiency of carnitine is known to have major deleterious effects on the CNS. Several syndromes of secondary carnitine deficiency have been described that may result from defects in intermediary metabolism and alterations principally involving mitochondrial oxidative pathways. Mitochondrial superoxide formation resulting from disturbed electron transfer within the respiratory chain may affect the activities of respiratory chain complexes I, II, III, IV, and V and underlie some CNS pathologies. This mitochondrial dysfunction may be ameliorated by L-C and its esters. In addition to its metabolic role, L-C and its esters such as acetyl-L-carnitine (ALC) poses unique neuroprotective, neuromodulatory, and neurotrophic properties which may play an important role in counteracting various disease processes. Neural dysfunction and metabolic imbalances underlie many diseases, and the inclusion of metabolic modifiers may provide an alternative and early intervention approach, which may limit further developmental damage, cognitive loss, and improve long term therapeutic outcomes. The neurophysiological and neuroprotective actions of L-C and ALC on cellular processes in the central and peripheral nervous system show such effects. Indeed, many studies have shown improvement in processes, such as memory and learning, and are discussed in this review. PMID- 15363641 TI - Carnitine: transport and physiological functions in the brain. AB - Carnitine (4-N-trimethylammonium-3-hydroxybutyric acid), a compound necessary for a transfer of fatty acids for their oxidation within the cell, accumulates in brain although beta-oxidation of fatty acids is very low in neurons. Carnitine accumulates to lower extent in the brain than in peripheral tissues and the mechanism of its transport through the blood-brain barrier is discussed, with the involvement of two transporters, OCTN2 and B(0,+) being presented. A limitation by the blood-brain barrier of carnitine supply for the brain and the mechanism of its transport to neural cells by a protein belonging to neurotransmitters' transporters superfamily is further discussed. Due to the beneficial effects of administration of acetylcarnitine in case of patients with dementia, the role of this acylcarnitine is presented in the context of neuronal cell metabolism and the role of acetylcarnitine in the synthesis of acetylcholine. The roles of long chain acyl derivatives of carnitine, in particular palmitoylcarnitine, responsible for interaction with the membranes, lipids acylation and specific interactions with proteins have been summarized. Stimulation of protein palmitoylation and a possibility of changing the acylation status of G proteins is described, as well as interaction of palmitoylcarnitine with protein kinase C. Diminished interaction of the isoform delta of this kinase with GAP-43 (B-50, neuromodulin), whose expression increases upon accumulation of either carnitine or palmitoylcarnitine points to a possible regulation of differentiation by these compounds and their role in neuroregeneration. PMID- 15363642 TI - Long-term effect of three different strategies for mass diethylcarbamazine administration in bancroftian filariasis: follow-up at 10 years after treatment. AB - The long-term effect of three different strategies for mass diethylcarbamazine (DEC) administration in bancroftian filariasis was assessed 10 years after start of treatment in three endemic communities in Tanzania. The strategies were the standard 12 day treatment (strategy I); a semi-annual single-dose treatment (strategy II); and a monthly low-dose treatment (strategy III). Treatment was given only during the first year. Following reductions immediately after treatment, overall community microfilaraemia levels were approaching pre treatment levels in all three communities, 10 years later. In individuals who were microfilaria-positive and treated at baseline, the treatment had a long-term effect on microfilarial intensities, with geometric mean intensities being only 11%, 13% and 2% of pre-treatment levels 10 years later for strategies I, II and III, respectively. This suppressive effect was most pronounced for strategy III, which also cleared microfilaraemia and circulating filarial antigenaemia in a larger proportion of treated individuals than the other strategies. Most of the follow-up individuals who developed microfilaraemia between 2 and 10 years after start of treatment had also been microfilaraemic before treatment, suggesting that reappearance of microfilaraemia may be due to surviving female worms and/or that previously microfilaraemic individuals have a higher chance of reinfection than previously amicrofilaraemic individuals. PMID- 15363643 TI - Case management of a multidrug-resistant Shigella dysenteriae serotype 1 outbreak in a crisis context in Sierra Leone, 1999-2000. AB - From December 1999 to the end of February 2000, 4218 cases of dysentery were reported in Kenema district, southeastern Sierra Leone, by a Medecins Sans Frontieres team operating in this region. Shigella dysenteriae serotype 1 was isolated from the early cases. The overall attack rate was 7.5% but higher among children under 5 years (11.2%) compared to the rest of the population (6.8%) (RR = 1.6; 95% CI 1.5-1.8). The case fatality ratio was 3.1%, and higher for children under 5 years (6.1% vs. 2.1%) (RR = 2.9; 95% CI 2.1-4.1). A case management strategy based on stratification of affected cases was chosen in this resource poor setting. Patients considered at higher risk of death were treated with a 5 day ciprofloxacin regimen in isolation centres. Five hundred and eighty-three cases were treated with a case fatality ratio of 0.9%. Patients who did not have signs of severity when seen by health workers were given hygiene advice and oral rehydration salts. This strategy was effective in this complex emergency. PMID- 15363644 TI - Field evaluation of the efficacy and persistence of insect repellents DEET, IR3535, and KBR 3023 against Anopheles gambiae complex and other Afrotropical vector mosquitoes. AB - Synthetic insect repellents, IR3535 and KBR 3023 (also known as picaridin, or by the trade name Bayrepel, were tested in Burkina Faso against mosquito vectors of disease to compare their relative efficacy and persistence profiles to those of the 'gold standard' DEET. Collection of >49000 mosquitoes (approximately 95% belonging to the Anopheles gambiae complex) showed that after an exposure of 10h, KBR 3023 produced the highest protection against anophelines, followed by DEET, then IR3535. The response of aedines was more variable. By fitting a logistic plane model we estimated 95% effective dosages (ED95) for An. gambiae s.l., as well as a decay constant characterizing the exponential loss of repellent from the skin, with time. The ED95 values for DEET, IR3535, and KBR 3023 were 94.3, 212.4, and 81.8 microg/cm2 respectively. The decay constants were estimated at 0.241, -0.240, and -0.170 h(-1) respectively. The corresponding estimates of half life were 2.9, 2.9, and 4.1h. Immunoenzymatic detection of the circumsporozoite protein (CSP) of Plasmodium falciparum in 842 An. gambiae s.l. showed that CSP positive mosquitoes were equally frequent in treated and control subjects, indicating that the repellents could produce a reduction in the number of malaria infectious bites. PMID- 15363645 TI - Prevalence and intensity of intestinal parasitic infections in relation to nutritional status in Mexican schoolchildren. AB - Undernutrition and intestinal parasitic infections affect childhood development and morbidity in many developing countries. Undernutrition may increase susceptibility to parasitic infections which in turn impair the nutritional status of the host. The relationship between intestinal parasitic infections and nutritional status in 400 Mexican schoolchildren was investigated. More than half of the children in the study showed intestinal parasites and polyparasitism. The prevalence of helminth infections was significantly higher in Oaxaca than in Sinaloa (P < 0.05). Z scores for weight-for-age (WA) and height-for-age (HA) were much lower in children of Oaxaca than in Sinaloa (P < 0.001). A significantly higher Z score for weight-for-height (WH), WA, and HA were found in non-infected versus infected children (P < 0.05). Higher prevalences of intestinal infections were found in children with lower HA and WA than in normally nourished children (P < 0.05). Higher intensities of Ascaris lumbricoides and Trichuris trichiura were found in the schoolchildren of Sinaloa than in Oaxaca (P < 0.01). Negative and significant associations were found between Hymenolepis nana and T. trichiura infection (eggs per gram) and nutritional status. Intestinal parasitic infections may be regarded as main risk factors associated with poor nutritional status in Mexican schoolchildren. PMID- 15363646 TI - Cytokine responses and progression to active tuberculosis in HIV-1-infected Ugandans: a prospective study. AB - Identifying correlates of immunity or susceptibility to disease promotes understanding of pathogenesis and development of diagnostic tools, treatments, and vaccines. There is evidence that type 1 cytokine responses are associated with protection against tuberculosis, and suppression of type 1, or switching to type 2 responses, with susceptibility, but this has not been studied prospectively. We studied a cohort of 631 HIV-1-infected Ugandan adults. At enrollment we performed whole blood cultures for type 1 (interferon [IFN]-gamma, interleukin [IL]-2) and type 2/immunosuppressive (IL-5, IL-10) responses to mycobacterial antigens (purified protein derivative [PPD] and culture filtrate proteins [CFP]). The incidence of tuberculosis was not associated with IFN-gamma responses, but was higher among participants with IL-2 responses (adjusted rate ratios [RR]: PPD 3.48; CFP 3.99; P < 0.001). For tuberculin skin test-positive participants, high incidence was also associated with an IL-10 response to PPD (adjusted RR 6.24, P = 0.03); for those with a BCG scar, high incidence was associated with positive IL-5 responses (adjusted RRs: PPD 3.64, P = 0.006; CFP 3.44, P = 0.04). The association with IL-2 production may reflect a response to tuberculous infection or to activating disease; the associations with IL-10 and IL-5 are in keeping with the expected role of immunosuppressive or type 2 cytokines. PMID- 15363647 TI - Impact of improved vaccination programme and intensive care facilities on incidence and outcome of tetanus in southern Vietnam, 1993-2002. AB - Unvaccinated individuals throughout the world are vulnerable to tetanus, but there are few data regarding the impact of focused vaccination programmes and modern intensive care facilities on the disease, particularly in the developing world. The Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam admitted 2422 patients with tetanus aged > or =1 year between April 1993 and December 2002, during which time vaccine coverage and treatment facilities improved. The proportion of children < or =10 years old admitted with tetanus fell from 11.1 to 5.6% over the 10 year period (P = 0.002). The proportion of women aged 20-40 years fell from 10.1 to 1.2% (P < 0.001). Mortality rates fell from a maximum of 27.81% in 1994 to 10.04% in 2002 (P < 0.001). Thus, a marked reduction in tetanus incidence has occurred in age groups specifically targeted by the national vaccination programme. However, tetanus continues to be a major cause of morbidity and mortality in individuals outside the target population. Improved intensive care facilities, such as mechanical ventilation and low-cost infection control procedures are associated with a significant reduction in mortality. PMID- 15363648 TI - The changing pattern of bloodstream infections associated with the rise in HIV prevalence in northeastern Thailand. AB - A survey of bloodstream infections was conducted in the large regional hospital in Ubon Ratchatani, northeastern Thailand between 1989 and 1998, during the onset of the HIV epidemic. The incidence of Staphylococcus aureus, Escherichia coli, Klebsiella/Enterobacter and Pseudomonas aeruginosa bacteraemias remained constant whereas infections caused by Burkholderia pseudomallei, non-typhoid Salmonellae, Cryptococcus neoformans, Penicillum marneffei and to a lesser extent Streptococcus pneumoniae all rose. Burkholderia pseudomallei infections were unrelated to HIV, whereas the other infections were associated directly with HIV. Group D non-typhoid Salmonellae bloodstream infections (mainly Salmonella enteritidis) rose coincident with the increase in HIV seroprevalence, and preceded the increase in the other HIV-associated infections. Other non-typhoid Salmonella bacteraemias increased two years after the rise in group D infections, and invasive yeast infections increased four years later, coincident with the increase in AIDS. Increasing Group D non-typhoid Salmonella bloodstream infections are an early warning signal of an impending rise in AIDS. PMID- 15363649 TI - Analysis of the force-sharing problem using an optimization model. AB - In biomechanics, one frequently used approach for finding a unique set of muscle forces in the 'force-sharing problem' is to formulate and solve a non-linear optimization problem of the form: min phi(f)= summation operator (f(i)/omega(i))(alpha) subject to Af = b and f > or = 0. Solutions to this problem have typically been obtained numerically for complex models, or analytically for specific musculoskeletal geometries. Here, we present simple geometrical methods for analyzing the solution to this family of optimization problems for a general n-degrees-of-freedom musculoskeletal system. For example, it is shown that the moment-arm vectors of active (f(i) > 0) and passive (f(i) = 0) muscles are separated by a hyperplane through the origin of the moment-arm vector space. For the special case of a system with two degrees-of-freedom, solutions can be readily represented in graphical form. This allows for powerful interpretations of force-sharing calculated using optimization. PMID- 15363650 TI - Stochastic optimal therapy for enhanced immune response. AB - Therapeutic enhancement of humoral immune response to microbial attack is addressed as the stochastic optimal control of a dynamic system. Without therapy, the modeled immune response depends upon the initial concentration of pathogens in a simulated attack. Immune response can be augmented by agents that kill the pathogen directly, that stimulate the production of plasma cells or antibodies, or that enhance organ health. Using a generic mathematical model of immune response to the infection (i.e., of the dynamic state of the system), previous papers demonstrated optimal (open-loop) and neighboring-optimal (closed-loop) control solutions that defeat the pathogen and preserve organ health, given initial conditions that otherwise would be lethal [Optimal Contr. Appl. Methods 23 (2002) 91, Bioinformatics 18 (2002) 1227]. Therapies based on separate and combined application of the agents were derived by minimizing a quadratic cost function that weighted both system response and drug usage, providing implicit control over harmful side effects. Here, we focus on the effects that corrupted or incomplete measurements of the dynamic state may have on neighboring-optimal feedback control. Imperfect measurements degrade the precision of feedback adjustments to therapy; however, optimal state estimation allows the feedback strategy to be implemented with incomplete measurements and minimizes the expected effects of measurement error. Complete observability of the perturbed state for this four state example is provided by measurement of four of the six possible pairs of two variables, either set of three variables, or all four variables. The inclusion of state estimation extends the applicability of optimal control theory for developing new therapeutic protocols to enhance immune response. PMID- 15363651 TI - On the development and application of a continuous-discrete recursive prediction error algorithm. AB - Recursive state and parameter reconstruction is a well-established field in control theory. In the current paper we derive a continuous-discrete version of recursive prediction error algorithm and apply the filter in an environmental and biological setting as a possible alternative to the well-known extended Kalman filter. The framework from which the derivation is started is the so-called 'innovations-format' of the (continuous time) system model, including (discrete time) measurements. After the algorithm has been motivated and derived, it is subsequently applied to hypothetical and 'real-life' case studies including reconstruction of biokinetic parameters and parameters characterizing the dynamics of a river in the United Kingdom. Advantages and characteristics of the method are discussed. PMID- 15363652 TI - Tumour eradication by antiangiogenic therapy: analysis and extensions of the model by Hahnfeldt et al. (1999). AB - The model proposed by Hahnfeldt et al. (1999) describes the growth of a tumour assuming that tumour growth is strictly controlled by the evolution of the vascular network that supplies oxygen and nutrients to tumour cells. Consequently, it provides a framework to represent the effects of antiangiogenic therapies. In this paper, some possible modifications of that model are proposed, and conditions that guarantee the eradication of the tumour under a regimen of periodic antiangiogenic therapy are derived. The model variants considered assume the potential doubling time of the vasculature to be constant, and subdivide the endothelial cell pool, which is involved in angiogenesis, in resting and proliferating cells allowing for a more detailed description of drug effects. PMID- 15363653 TI - An approach to the residence time distribution for stochastic multi-compartment models. AB - Stochastic compartmental models are widely used in modeling processes such as drug kinetics in biological systems. This paper considers the distribution of the residence times for stochastic multi-compartment models, especially systems with non-exponential lifetime distributions. The paper first derives the moment generating function of the bivariate residence time distribution for the two compartment model with general lifetimes and approximates the density of the residence time using the saddlepoint approximation. Then, it extends the distributional approach to the residence time for multi-compartment semi-Markov models combining the cofactor rule for a single destination and the analytic approach to the two-compartment model. This approach provides a complete specification of the residence time distribution based on the moment generating function and thus facilitates an easier calculation of high-order moments than the approach using the coefficient matrix. Applications to drug kinetics demonstrate the simplicity and usefulness of this approach. PMID- 15363654 TI - Mathematical analysis of competition between sensory ganglion cells for neurotrophic factor in the skin. AB - A model is presented of competition between sensory axons for trophic molecules (e.g. a neurotrophin such as NGF), produced in a region of skin small enough to permit their free diffusion throughout it; e.g., a touch dome, or a vibrissal follicle hair sinus. The variables specified are the number of high affinity trophic factor receptors per axon terminal and the concentration of trophic factor in the extracellular space. Previous models of this class predicted the loss of all the axons innervating the region except the one requiring least trophic factor for its maintenance, even with high rates of trophic factor production. In the present model, we have imposed upper limits to axonal growth, thereby introducing new equilibria, and we show by a global analysis using LaSalle's theorem, and also by local analysis, that several axons can then coexist if the rate of production of trophic molecules is sufficiently high. PMID- 15363655 TI - Lipid rafts in neuregulin signaling at synapses. AB - Neuregulins are a family of EGF domain-containing factors that play an important role in development. In the nervous system, they promote glial differentiation, induce neurotransmitter receptor expression, and regulate synaptic plasticity. Recent studies indicate that ErbB protein tyrosine kinases, neuregulin receptors, translocate to lipid raft microdomains in the plasma membrane in response to neuregulin. Localization of ErbB proteins in lipid rafts appeared to be necessary for neuregulin signaling and regulation of synaptic plasticity. We will review recent studies of lipid rafts and neuregulin function and discuss possible roles of lipid rafts in compartmentalized neuregulin signaling and translocation of ErbB proteins to synapses. PMID- 15363656 TI - Antidiabetic effect of Pycnogenol French maritime pine bark extract in patients with diabetes type II. AB - A double-blind, placebo-controlled, randomized, multi-center study was performed with 77 diabetes type II patients to investigate anti-diabetic effects of the French maritime pine bark extract, Pynogenol. Supplementation with 100 mg Pycnogenol for 12 weeks, during which a standard anti-diabetic treatment was continued, significantly lowered plasma glucose levels as compared to placebo. HbA1(c) was also lowered; however, the difference as compared to placebo was statistically significant only for the first month. In the Pycnogenol-group endothelin-1 was significantly decreased, while 6-ketoprostaglandin F(1a) in plasma was elevated compared to placebo. Nitric oxide levels in plasma increased during treatment in both groups, but, differences did not reach statistical significance. Pycnogenol was well-tolerated with ECG, electrolytes, creatinine and blood urea nitrogen remaining unchanged in both groups. Mild and transient unwanted effects were reported for both groups without significant differences. Supplementation of Pycnogenol to conventional diabetes treatment lowers glucose levels and improves endothelial function. PMID- 15363657 TI - Capsazepine elevates intracellular Ca2+ in human osteosarcoma cells, questioning its selectivity as a vanilloid receptor antagonist. AB - Capsazepine is thought to be a selective antagonist of vanilloid type 1 receptors; however, its other in vitro effect on different cell types is unclear. In human MG63 osteosarcoma cells, the effect of capsazepine on intracellular Ca(2+) concentrations ([Ca(2+)](i)) and cytotoxicity was explored by using fura-2 and tetrazolium, respectively. Capsazepine caused a rapid rise in [Ca(2+)](i) in a concentration-dependent manner with an EC(50) value of 100 microM. Capsazepine induced [Ca(2+)](i) rise was partly reduced by removal of extracellular Ca(2+), suggesting that the capsazepine-induced [Ca(2+)](i) rise was composed of extracellular Ca(2+) influx and intracellular Ca(2+). In Ca(2+)-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca(2+)-ATPase, caused a monophasic [Ca(2+)](i) rise, after which the increasing effect of capsazepine on [Ca(2+)](i) was inhibited by 75%. Conversely, pretreatment with capsazepine to deplete intracellular Ca(2+) stores totally prevented thapsigargin from releasing more Ca(2+). U73122, an inhibitor of phospholipase C, abolished histamine (an inositol 1,4,5-trisphosphate-dependent Ca(2+) mobilizer)-induced, but not capsazepine-induced, [Ca(2+)](i) rise. Overnight treatment with 1-100 microM capsazepine inhibited cell proliferation in a concentration-dependent manner. These findings suggest that in human MG63 osteosarcoma cells, capsazepine increases [Ca(2+)](i) by stimulating extracellular Ca(2+) influx and also by causing intracellular Ca(2+) release from the endoplasmic reticulum via a phospholiase C-independent manner. Capsazepine may be mildly cytotoxic. PMID- 15363658 TI - Preemptive effects of intrathecal cyclooxygenase inhibitor or nitric oxide synthase inhibitor on thermal hypersensitivity following peripheral nerve injury. AB - The present study provides an important implication for the management of chronic neuropathic pain, focusing on prostaglandin (PG) and nitric oxide (NO) in the spinal cord. To determine if spinally administered cyclooxygenase (COX) inhibitor or nitric oxide synthase (NOS) inhibitor had preemptive analgesia on thermal hypersensitivity induced by chronic constrictive nerve injury, Sprague-Dawley rats were chronically implanted with an intrathecal (i.t.) catheter. The left sciatic nerve was loosely ligated with 2-mm polyethylene tubing to produce painful mononeuropathy. Animals received tenoxicam (7.5, 15 or 30 micromol/10 microl, i.t.), NS-398 (15 or 30 micromol), or L-NAME (30, 150 or 300 micromol) immediately before the nerve injury, followed by daily injection extending into the four postoperative days. The hindpaw was immersed into a hot (42 degrees C, 44 degrees C and 46 degrees C) or cold (10 degrees C) water bath. The paw immersion test revealed significant thermal hyperalgesia and allodynia 5 day after nerve injury in vehicle control animals. Tenoxicam (7.5, 15 or 30 micromol) or L-NAME (30, 150 or 300 micromol) dose-dependently attenuated hyperalgesia and allodynia. Equimolar dose of NS-398 (15 or 30 micromol) also diminished these nociceptive behaviors. Higher dose of either drug primarily produced longer duration of inhibition. The inhibitory effect of tenoxicam (30 micromol) on hyperalgesia was more effective than that of an equimolar dose of NS-398 or L NAME. These results demonstrated that intrathecally administered COX inhibitor or NOS inhibitor provides preemptive analgesia on thermal hypersensitivity following chronic constrictive nerve injury in rats. PMID- 15363659 TI - Oxidative stress in autism: increased lipid peroxidation and reduced serum levels of ceruloplasmin and transferrin--the antioxidant proteins. AB - Autism is a neurological disorder of childhood with poorly understood etiology and pathology. We compared lipid peroxidation status in the plasma of children with autism, and their developmentally normal non-autistic siblings by quantifying the levels of malonyldialdehyde, an end product of fatty acid oxidation. Lipid peroxidation was found to be elevated in autism indicating that oxidative stress is increased in this disease. Levels of major antioxidant proteins namely, transferrin (iron-binding protein) and ceruloplasmin (copper binding protein) in the serum, were significantly reduced in autistic children as compared to their developmentally normal non-autistic siblings. A striking correlation was observed between reduced levels of these proteins and loss of previously acquired language skills in children with autism. These results indicate altered regulation of transferrin and ceruloplasmin in autistic children who lose acquired language skills. It is suggested that such changes may lead to abnormal iron and copper metabolism in autism, and that increased oxidative stress may have pathological role in autism. PMID- 15363660 TI - Effects of cimetidine-like drugs on recombinant GABAA receptors. AB - Even though conventional systemic doses of cimetidine and other histamine H(2) antagonists display minimal brain penetration, central nervous system (CNS) effects (including seizures and analgesia) have been reported after administration of these drugs in animals and man. To test the hypothesis that cimetidine-like drugs produce these CNS effects via inhibition of GABA(A) receptors, the actions of these drugs were studied on seven different, precisely defined rat recombinant GABA(A) receptors using whole-cell patch clamp recordings. The H(2) antagonists famotidine and tiotidine produced competitive and reversible inhibition of GABA-evoked currents in HEK293 cells transfected with various GABA(A) receptor subunits (IC(50) values were between 10-50 microM). In contrast, the H(2) antagonist ranitidine and the cimetidine congener improgan had very weak (if any) effects (IC(50) > 50 microM). Since the concentrations of cimetidine-like drugs required to inhibit GABA(A) receptors in vitro (greater than 50 microM) are considerably higher than those found during analgesia and/or seizures (1-2 microM), the present results suggest that cimetidine-like drugs do not appear to produce seizures or analgesia by directly inhibiting GABA(A) receptors. PMID- 15363661 TI - Flavin-containing monooxygenase activity can be inhibited by nitric oxide mediated S-nitrosylation. AB - Nitric oxide (NO) modifies the functions of a variety of proteins containing cysteine thiols or transition-metal centers, particularly by S-nitrosylation. In inflamed liver, NO is overproduced and hepatic drug-metabolizing enzymes, the flavin-containing monooxygenases (FMOs) and cytochrome P450s (CYPs), are suppressed. However, the NO-related mechanisms underlying the loss of these activities are not well understood, particularly for FMOs. In this study, we suggest that FMO3, the major FMO in human liver, is modified post-translationally by NO. This hypothesis is based on the imbalance observed between the decrease in FMO3 expression (40.7% of controls) and FMO3-specific ranitidine N-oxidation activity (15.1%), and on the partial or complete reversibility of FMO inhibition by sulfhydryl-reducing regents such as DTT (effective on both S-S and S-NO adducts) and ascorbate (effective on S-NO only). Furthermore, NO donors (SNP, SNAP, and Sin-1), including the pure NO donor DEA/NO, directly suppressed in vitro FMO activity (N- or S-oxidation of ranitidine, trimethylamine, and thiobenzamide) in human liver microsomal proteins and recombinant human FMO3. These activities were restored completely after treatment with DTT or ascorbate. These results suggest that NO-mediated S-nitrosylation is involved in the rigorous inhibition of FMO activity in vitro and in vivo, resulting in the suppression of FMO-based drug metabolism or detoxification. PMID- 15363662 TI - Nitric oxide-dependent synthesis of vascular endothelial growth factor is impaired by high glucose. AB - Synthesis of vascular endothelial growth factor (VEGF), the major angiogenic molecule, is induced by nitric oxide (NO) in various cell types, including vascular smooth muscle cells (VSMC). Therefore, compounds which inhibit NO generation can also influence VEGF synthesis. Here we investigated the effect of increased glucose concentration (25 mM vs. 5.5 mM) on cytokine-induced VEGF synthesis in rat VSMC. The cells growing in the medium containing 5.5 mM glucose and exposed to IL-1-beta, TNF-alpha and IFN-gamma induced expression of an inducible isoform of nitric oxide synthase (NOS II). This is followed by generation of NO and the concomitant expression of VEGF gene and release of VEGF protein. In contrast, 25 mM glucose impaired induction of NOS II expression and thus NO synthesis was lower than in 5.5 mM glucose. Consequently, the VEGF promoter activation was attenuated, resulting in decreased mRNA synthesis and lower production of VEGF protein. The results indicate that abnormally high concentrations of glucose can impair generation of NO and the NO-dependent VEGF synthesis. This may play a role in the development and progression of vascular dysfunctions in cardiovascular diseases. PMID- 15363663 TI - Intermittent hypoxia protects the rat heart against ischemia/reperfusion injury by activating protein kinase C. AB - The aim of this study was to investigate whether and how protein kinase C (PKC) was involved in the protection afforded by intermittent hypoxia (IH) and the subcellular distribution of different PKC isozymes in rat left ventricle. Post ischemic recovery of left ventricular developed pressure and +/-dP/dtmax in IH hearts were higher than those of normoxic hearts. Chelerythrine (CHE, 5 microM), a PKC antagonist, significantly inhibited the protective effects of IH, but had no influence on normoxic hearts. CHE significantly reduced the effect of IH on the time to maximal contracture (Tmc), but had no significant effect on the amplitude of maximal contracture (Amc) in IH group. In isolated normoxic cardiomyocytes, [Ca(2+)](i), measured as arbitrary units of fluorescence ratio (340 nm/380 nm) of fura-2, gradually increased during 20 min simulated ischemia and kept at high level during 30 min reperfusion. However, [Ca(2+)](i) kept at normal level during simulated ischemia and reperfusion in isolated IH cardiomyocytes. In normoxic myocytes, [Na(+)](i), indicated as actual concentration undergone calibration, gradually increased during 20 min simulated ischemia and quickly declined to almost the same level as that of pre-ischemia during 30 min simulated reperfusion. However, in IH myocytes, [Na(+)](i) increased to a level lower than the corresponding of normoxic myocytes during simulated ischemia and gradually reduced to the similar level as that of normoxic myocytes after simulated reperfusion. 5 microM CHE greatly increased the levels of [Ca(2+)](i) and [Na(+)](i) during ischemia and reperfusion in normoxic and IH myocytes. In addition, we demonstrated that IH up-regulated the baseline protein expression of particulate fraction of PKC-alpha, epsilon, delta isozymes. There is no significant difference of protein expression of PKC-alpha, epsilon, delta isozymes in cytosolic fraction between IH and normoxic group. The above results suggested that PKC contributed to the cardioprotection afforded by IH against ischemia/reperfusion (I/R) injury; the basal up-regulation of the particulate fraction of PKC-alpha, epsilon, delta isozymes in IH rat hearts and the contribution of PKC to the elimination of calcium and sodium overload might underlie the mechanisms of cardioprotection by IH. PMID- 15363664 TI - Doxazosin treatment causes differential alterations of alpha 1-adrenoceptor subtypes in the rat kidney, heart and aorta. AB - We have previously demonstrated that long-term administration of doxazosin, an alpha(1)-adrenoceptor (alpha(1)-AR) antagonist, causes an up-regulation in the expression of alpha(1)-AR subtype mRNAs in the rat genitourinary tract and suggested that these changes may affect long-term effectiveness of alpha(1)-AR antagonists when used to treat the lower urinary tract symptoms of benign prostatic hyperplasia. As chronic administration of alpha(1)-AR antagonists may cause similar alterations in other tissues in which alpha(1)-ARs play a physiologic role, we examined the effects of long-term administration of doxazosin on the expression of alpha(1)-AR subtype mRNAs in several rat tissues. Rats were treated with doxazosin (4 mg/kg/day subcutaneously, supplemented with 4 mg/kg/day orally) for 12 weeks. The cDNA was prepared by reverse transcription of RNA extracted from the rat kidney, heart and aorta. alpha(1A), alpha(1B) and alpha(1D)-AR mRNA expression levels were quantified by real-time reverse transcription polymerase chain reaction. The rank order of expression levels of the alpha(1)-AR mRNAs in rat tissues were: alpha(1A)-AR, kidney > heart > aorta; alpha(1B)-AR, heart > kidney > aorta; alpha(1D)-AR, aorta > kidney = heart. Chronic administration of doxazosin caused an up-regulation in the mRNA level of alpha(1A), alpha(1B) and alpha(1D)-ARs in the rat kidney, heart and aorta, respectively. Our data demonstrate that doxazosin treatment causes differential alterations in the expression of alpha(1)-AR subtype mRNAs in different rat tissues. These findings may provide insight into the long-term effects of alpha(1)-AR antagonists in the treatment of diseases involving tissues whose function is regulated by alpha(1)-ARs. PMID- 15363665 TI - Arthur T. Winfree (1942-2002). PMID- 15363666 TI - Art Winfree, artist of science. PMID- 15363667 TI - Art Winfree and the bidomain model of cardiac tissue. AB - This paper reviews Art Winfree's contributions to the bidomain model of cardiac tissue. Specifically, he first predicted quatrefoil reentry, he showed that an S1 refractory gradient is not required for an S2 stimulus to induce reentry, and his work on spiral wave meandering led to studies on how the path of the tip of a spiral wave is influenced by tissue anisotropy. PMID- 15363668 TI - Clocks and patterns in myxobacteria: a remembrance of Art Winfree. AB - At the beginning of their aggregation phase waves of cell density sweep across the surface of myxobacteria colonies. These waves are unlike any other in biology. Waves can be linear, concentric or spiral and when they collide, instead of annihilating one another they appear to pass through each other unchanged. Moreover, the wavelength determines the spacing and pattern of fruiting bodies that will rise up presaging sporulation. The explanation for these waves was suggested by the work of Art Winfree on cellular clocks, and confirmed by a mathematical model that explains all of the observed wave behavior. The story of how this model evolved illustrates the roles of chance and scientific networking in the search for the explanation of a new phenomenon. PMID- 15363669 TI - The topology of defibrillation. AB - We describe how Art Winfree's ideas about phase singularities can be used to understand the response of cardiac tissue with a random preexisting pattern of reentrant waves (fibrillation) to a large brief current stimulus. This discussion is organized around spatial dimension, beginning with a discussion of reentry on a periodic ring, followed by reentry in a two-dimensional planar domain (spiral waves), and ending with consideration of three-dimensional reentrant patterns (scroll waves). In all cases, we show how reentrant activity is changed by the application of a shock, describing conditions under which defibrillation is successful or not. Using topological arguments we draw the general conclusion that with a generic placement of stimulating electrodes, large-scale virtual electrodes do not give an adequate explanation for the mechanism of defibrillation. PMID- 15363670 TI - Molecular mechanisms and global dynamics of fibrillation: an integrative approach to the underlying basis of vortex-like reentry. AB - Art Winfree's scientific legacy has been particularly important to our laboratory whose major goal is to understand the mechanisms of ventricular fibrillation (VF). Here, we take an integrative approach to review recent studies on the manner in which nonlinear electrical waves organize to result in VF. We describe the contribution of specific potassium channel proteins and of the myocardial fiber structure to such organization. The discussion centers on data derived from a model of stable VF in the Langendorff-perfused guinea pig heart that demonstrates distinct patterns of organization in the left (LV) and right (RV) ventricles. Analysis of optical mapping data reveals that VF excitation frequencies are distributed throughout the ventricles in clearly demarcated domains. The highest frequency domains are found on the anterior wall of the LV at a location where sustained reentrant activity is present. The optical data suggest that a high frequency rotor that remains stationary in the LV is the mechanism that sustains VF in this model. Computer simulations predict that the inward rectifying potassium current (IK1) is an essential determinant of rotor stability and frequency, and patch-clamp results strongly suggest that the outward component of IK1 of cells in the LV is significantly larger than in the RV. Additional computer simulations and analytical procedures predict that the filaments of the reentrant activity (scroll waves) adopt a non-random configuration depending on fiber organization within the ventricular wall. Using the minimal principle we have concluded that filaments align with the trajectory of least resistance (i.e. the geodesic) between their endpoints. Overall, the data discussed have opened new and potentially exciting avenues of research on the possible role played by inward rectifier channels in the mechanism of VF, as well as the organization of its reentrant sources in three-dimensional cardiac muscle. Such an integrative approach may lead us toward an understanding of the molecular and structural basis of VF and hopefully to new preventative approaches. PMID- 15363671 TI - A physical approach to remove anatomical reentries: a bidomain study. AB - Controlling cardiac chaos is often achieved by applying a large damaging electric shock-defibrillation. It removes all waves, without differentiating reentries and normal waves, anatomical and functional reentries. Anatomical reentries can be removed by anti-tachycardia pacing (ATP) as well. But ATP requires the knowledge of the position of the reentry, and an access to it with an invasive stimulating electrode. We show that the physics of electric field distribution between cardiac cells permits one to deliver an electric pulse exactly to the core of an anatomical reentry, without knowing its position and even to locations where access with a stimulating electrode is not possible. The energy needed is two orders of magnitude less than defibrillation energy. The results are insensitive to both a detailed ionic model and to the geometry of the fibers. PMID- 15363672 TI - Apparent discontinuities in the phase-resetting response of cardiac pacemakers. AB - Injection of a brief stimulus pulse resets the spontaneous periodic activity of a sinoatrial node cell: a stimulus delivered early in the cycle generally delays the time of occurrence of the next action potential, while the same stimulus delivered later causes an advance. We investigate resetting in two models, one with a slow upstroke velocity and the other with a fast upstroke velocity, representing central and peripheral nodal cells, respectively. We first formulate each of these models as a classic Hodgkin-Huxley type of model and then as a model representing a population of single channels. In the Hodgkin-Huxley-type model of the slow-upstroke cell the transition from delay to advance is steep but continuous. In the corresponding single-channel model, due to the channel noise then present, repeated resetting runs at a fixed stimulus timing within the transitional range of coupling intervals lead to responses that span a range of advances and delays. In contrast, in the fast-upstroke model the transition from advance to delay is very abrupt in both classes of model, as it is in experiments on some cardiac preparations ("all-or-none" depolarization). We reduce the fast upstroke model from the original seven-dimensional system to a three-dimensional system. The abrupt transition occurs in this reduced model when a stimulus transports the state point to one side or the other of the stable manifold of the trajectory corresponding to the eigendirection associated with the smaller of two positive eigenvalues. This stable manifold is close to the slow manifold, and so canard trajectories are seen. Our results demonstrate that the resetting response is fundamentally continuous, but extremely delicate, and thus suggest one way in which one can account for experimental discontinuities in the resetting response of a nonlinear oscillator. PMID- 15363673 TI - Starting, stopping, and resetting biological oscillators: in search of optimum perturbations. AB - Biological oscillators are commonly subjected to a single brief stimulus to perturb the ongoing rhythm. After stimulation, the oscillator can recover although its phase may be advanced or delayed relative to the original cycle. A single transient perturbation can also stop the ongoing rhythm. Arthur Winfree systematically classified these responses to brief perturbations, as determined by the strength of the stimulus, and the phase within the cycle at which the stimulus was given. A natural question arises from Winfree's work. Are certain stimulus shapes better than others at achieving a desired effect? The present study explores this question using: (1) analysis of phase space geometry, (2) calculus of variations, and (3) analysis of responses to noisy perturbations. Methods 1 and 2 can yield exact solutions, but have limited applicability in biology because they require a detailed mathematical description of the oscillator. Method 3 is applicable to any oscillator without mathematical prerequisites. We validate this method by finding optimum stimuli that start and stop repetitive firing in a neural pacemaker model, and the optimum light stimulus for resetting the circadian rhythm in a model of the human circadian clock. We propose that the elucidation of optimum stimulus shapes may be useful for studying many periodic phenomena in biology and medicine. Optimum stimuli can be used to induce a desired behavior without producing undesirable side effects. PMID- 15363674 TI - Model based conjectures on mammalian clock controversies. AB - We explore some predictions of a previously developed detailed model of molecular timekeeping in mammals (Forger and Peskin, PNAS, 100:14806) in areas where our understanding of clock mechanisms are incomplete. It is conjectured that: (1) the clock's 24-h period depends on mRNA stability. (2) REV-ERBalpha suppresses and/or entrains rhythms in peripheral tissues by regulating CRY1 transcription. (3) CLK:BMAL1 oscillations are suppressed in the suprachiasmatic nuclei to enhance oscillations in other proteins. (4) PER2 is ineffective at causing phase advances because it is not induced by light during the late night. (5) The clock is a limit cycle oscillator that shows characteristics of the evening and morning oscillator model. PMID- 15363675 TI - Modeling the mammalian circadian clock: sensitivity analysis and multiplicity of oscillatory mechanisms. AB - We extend the study of a computational model recently proposed for the mammalian circadian clock (Proc. Natl Acad. Sci. USA 100 (2003) 7051). The model, based on the intertwined positive and negative regulatory loops involving the Per, Cry, Bmal1, and Clock genes, can give rise to sustained circadian oscillations in conditions of continuous darkness. These limit cycle oscillations correspond to circadian rhythms autonomously generated by suprachiasmatic nuclei and by some peripheral tissues. By using different sets of parameter values producing circadian oscillations, we compare the effect of the various parameters and show that both the occurrence and the period of the oscillations are generally most sensitive to parameters related to synthesis or degradation of Bmal1 mRNA and BMAL1 protein. The mechanism of circadian oscillations relies on the formation of an inactive complex between PER and CRY and the activators CLOCK and BMAL1 that enhance Per and Cry expression. Bifurcation diagrams and computer simulations nevertheless indicate the possible existence of a second source of oscillatory behavior. Thus, sustained oscillations might arise from the sole negative autoregulation of Bmal1 expression. This second oscillatory mechanism may not be functional in physiological conditions, and its period need not necessarily be circadian. When incorporating the light-induced expression of the Per gene, the model accounts for entrainment of the oscillations by light-dark (LD) cycles. Long-term suppression of circadian oscillations by a single light pulse can occur in the model when a stable steady state coexists with a stable limit cycle. The phase of the oscillations upon entrainment in LD critically depends on the parameters that govern the level of CRY protein. Small changes in the parameters governing CRY levels can shift the peak in Per mRNA from the L to the D phase, or can prevent entrainment. The results are discussed in relation to physiological disorders of the sleep-wake cycle linked to perturbations of the human circadian clock, such as the familial advanced sleep phase syndrome or the non-24h sleep wake syndrome. PMID- 15363676 TI - A model for restriction point control of the mammalian cell cycle. AB - Inhibition of protein synthesis by cycloheximide blocks subsequent division of a mammalian cell, but only if the cell is exposed to the drug before the "restriction point" (i.e. within the first several hours after birth). If exposed to cycloheximide after the restriction point, a cell proceeds with DNA synthesis, mitosis and cell division and halts in the next cell cycle. If cycloheximide is later removed from the culture medium, treated cells will return to the division cycle, showing a complex pattern of division times post-treatment, as first measured by Zetterberg and colleagues. We simulate these physiological responses of mammalian cells to transient inhibition of growth, using a set of nonlinear differential equations based on a realistic model of the molecular events underlying progression through the cell cycle. The model relies on our earlier work on the regulation of cyclin-dependent protein kinases during the cell division cycle of yeast. The yeast model is supplemented with equations describing the effects of retinoblastoma protein on cell growth and the synthesis of cyclins A and E, and with a primitive representation of the signaling pathway that controls synthesis of cyclin D. PMID- 15363677 TI - A proposal for using the ensemble approach to understand genetic regulatory networks. AB - Understanding the genetic regulatory network comprising genes, RNA, proteins and the network connections and dynamical control rules among them, is a major task of contemporary systems biology. I focus here on the use of the ensemble approach to find one or more well-defined ensembles of model networks whose statistical features match those of real cells and organisms. Such ensembles should help explain and predict features of real cells and organisms. More precisely, an ensemble of model networks is defined by constraints on the "wiring diagram" of regulatory interactions, and the "rules" governing the dynamical behavior of regulated components of the network. The ensemble consists of all networks consistent with those constraints. Here I discuss ensembles of random Boolean networks, scale free Boolean networks, "medusa" Boolean networks, continuous variable networks, and others. For each ensemble, M statistical features, such as the size distribution of avalanches in gene activity changes unleashed by transiently altering the activity of a single gene, the distribution in distances between gene activities on different cell types, and others, are measured. This creates an M-dimensional space, where each ensemble corresponds to a cluster of points or distributions. Using current and future experimental techniques, such as gene arrays, these M properties are to be measured for real cells and organisms, again yielding a cluster of points or distributions in the M dimensional space. The procedure then finds ensembles close to those of real cells and organisms, and hill climbs to attempt to match the observed M features. Thus obtains one or more ensembles that should predict and explain many features of the regulatory networks in cells and organisms. PMID- 15363678 TI - Netscan: a procedure for generating reaction networks by size. AB - In this paper, we describe an algorithm which can be used to generate the collection of networks, in order of increasing size, that are compatible with a list of chemical reactions and that satisfy a constraint. Our algorithm has been encoded and the software, called Netscan, can be freely downloaded from ftp://ftp.stat.ubc.ca/pub/riffraff/Netscanfiles, along with a manual, for general scientific use. Our algorithm may require pre-processing to ensure that the quantities it acts on are physically relevant and because it outputs sets of reactions, which we call canonical networks, that must be elaborated into full networks. PMID- 15363679 TI - Guidelines on the number of embryos transferred. AB - Based on data from the most current published CDC/ASRM/SART yearly report, guidelines for the number of embryos to be transferred in IVF cycles have been further refined. PMID- 15363680 TI - Guidelines for advertising by ART programs. AB - To protect the patient from getting misleading information, these guidelines have been revised to reflect more stringent requirements for advertising and marketing. PMID- 15363681 TI - Revised minimum standards for practices offering assisted reproductive technologies. PMID- 15363682 TI - 2002 guidelines for gamete and embryo donation: a practice committee report: guidelines and minimum standards. PMID- 15363683 TI - Guidelines for sperm donation. PMID- 15363684 TI - Guidelines for oocyte donation. PMID- 15363685 TI - Guidelines for cryopreserved embryo donation. PMID- 15363686 TI - Psychological assessment of gamete donors and recipients. PMID- 15363687 TI - Psychological guidelines for embryo donation. PMID- 15363688 TI - Appendix A: minimal genetic screening for gamete donors. PMID- 15363689 TI - Guidelines for the provision of infertility services. PMID- 15363690 TI - Hormonal contraception: recent advances and controversies. AB - This document will outline new delivery systems and contraceptive formulations, summarize recent advances in emergency contraception, and review the effects of hormonal contraception on cancer risks, cardiovascular disease, and bone. PMID- 15363691 TI - Current evaluation of amenorrhea. AB - Amenorrhea is the absence or abnormal cessation of the menses. Primary and secondary amenorrhea describe the occurrence of amenorrhea before and after menarche, respectively. PMID- 15363692 TI - Endometriosis and infertility. AB - Women with endometriosis typically present with pelvic pain, infertility or an adnexal mass. Surgery for persistent adnexal masses may be indicated to remove an endometrioma or other pelvic pathology. Surgical or medical therapy is efficacious for pelvic pain due to endometriosis, but treatment of endometriosis in the female partner of an infertile couple raises a number of complex clinical questions that do not have simple answers. PMID- 15363693 TI - Treatment of androgen deficiency in the aging male. AB - Although guidelines for androgen replacement therapy for older men have been developed by several groups, the data to support treatment are based on the results of only a small number of clinical trials. It has not been definitively established that the decline in testosterone seen in most aging men results in an androgen deficient state with health-related outcomes that can be improved by androgen therapy. PMID- 15363694 TI - Correct coding for laboratory procedures during assisted reproductive technology cycles. AB - New Current Procedural Terminology (CPT) codes for 2004 have been adopted for utilization with assisted reproductive technologies. It is important for professionals in the field of assisted reproductive technology (ART) to become familiar with them prior to their implementation. This document replaces the July 2002 American Society for Reproductive Medicine (ASRM) Practice Guideline Correct Coding for Laboratory Procedures During Assisted Reproductive Technology. PMID- 15363695 TI - Interpretation of clinical trial results. AB - This educational bulletin provides background and tips on how to recognize quality trials and then focuses on evaluating the validity, importance, and relevance of clinical trial results. PMID- 15363696 TI - Smoking and infertility. AB - Approximately 30% of reproductive age women and 35% of reproductive age men in the United States smoke cigarettes. Substantial harmful effects of cigarette smoke on fecundity and reproduction have become apparent but are not generally appreciated. PMID- 15363697 TI - Society for assisted reproductive technology position statement on donor suitability of recipients of smallpox vaccine (vaccinia virus). AB - Although there is presently no definitive evidence linking vaccinia virus transmission through reproductive cells, SART/ASRM accordingly recommends that assisted reproductive technology (ART) practitioners consider deferring donors who have recently received smallpox vaccine or contracted symptomatic vaccinia virus infection through close contact with a vaccine recipient (until after the vaccine or infectious scab has spontaneously separated). Good donor practice further suggests that donors who are not in good health, including those with recent complications from smallpox vaccine, should be similarly deferred. PMID- 15363698 TI - Estrogen and progestogen therapy in postmenopausal women. PMID- 15363699 TI - Ovarian hyperstimulation syndrome. PMID- 15363700 TI - Round spermatid nucleus injection (ROSNI). PMID- 15363701 TI - Use of clomiphene citrate in women. PMID- 15363702 TI - Information on commonly asked questions about genetic evaluation and counseling for infertile couples. PMID- 15363703 TI - Aging and infertility in women. PMID- 15363704 TI - The menopausal transition. PMID- 15363705 TI - Myomas and reproductive function. PMID- 15363706 TI - Salpingectomy for hydrosalpinx prior to in vitro fertilization. PMID- 15363707 TI - Preimplantation genetic diagnosis. PMID- 15363708 TI - Report on optimal evaluation of the infertile male. PMID- 15363709 TI - Report on evaluation of the azoospermic male. PMID- 15363710 TI - Report on management of obstructive azoospermia. PMID- 15363711 TI - Report on varicocele and infertility. PMID- 15363712 TI - Early diagnosis and management of ectopic pregnancy. PMID- 15363713 TI - Blastocyst production and transfer in clinical assisted reproduction. PMID- 15363714 TI - Does intracytoplasmic sperm injection (ICSI) carry inherent genetic risks? PMID- 15363715 TI - Multiple pregnancy associated with infertility therapy. PMID- 15363716 TI - Repetitive oocyte donation. PMID- 15363717 TI - Effectiveness and treatment for unexplained infertility. PMID- 15363718 TI - The role of assisted hatching in in vitro fertilization: A review of the literature. PMID- 15363719 TI - Status of environment and dietary estrogens--are they significant estrogens? PMID- 15363720 TI - Optimal evaluation of the infertile female. PMID- 15363721 TI - The evaluation and treatment of androgen excess. PMID- 15363722 TI - Use of insulin sensitizing agents in the treatment of polycystic ovary syndrome. PMID- 15363723 TI - Antiphospholipid antibodies do not affect IVF success. PMID- 15363724 TI - New techniques for sperm acquisition in obstructive azoospermia. PMID- 15363725 TI - Vasectomy reversal. PMID- 15363726 TI - Intravenous immunoglobulin (IVIG) and recurrent spontaneous pregnancy loss. PMID- 15363727 TI - Position statement on nurses performing limited ultrasound in a gyn/infertility setting. PMID- 15363728 TI - Elements to be considered in obtaining informed consent for ART. PMID- 15363729 TI - Electroejaculation (EEJ). PMID- 15363730 TI - Intracytoplasmic sperm injection (ICSI). PMID- 15363731 TI - Definition of "infertility". PMID- 15363732 TI - Definition of "experimental". PMID- 15363733 TI - Child-rearing ability and the provision of fertility services. AB - Fertility programs may withhold services when there are reasonable grounds for thinking that patients will not provide adequate child-rearing to offspring but are not obligated to do so. PMID- 15363734 TI - Informing offspring of their conception by gamete donation. AB - The Ethics Committee supports disclosure from parents to offspring about the use of donor gametes in their conception. The merits of further disclosure are discussed, and suggestions are made for policies for programs and sperm banks. PMID- 15363735 TI - Family members as gamete donors and surrogates. PMID- 15363736 TI - Donating spare embryos for embryonic stem-cell research. PMID- 15363737 TI - Human immunodeficiency virus and infertility treatment. PMID- 15363738 TI - Preconception gender selection for nonmedical reasons. PMID- 15363739 TI - Human somatic cell nuclear transfer (cloning). PMID- 15363740 TI - Financial incentives in recruitment of oocyte donors. PMID- 15363741 TI - Sex selection and preimplantation genetic diagnosis. PMID- 15363742 TI - Shared-risk or refund programs in assisted reproduction. PMID- 15363743 TI - Informed consent and the use of gametes and embryos for research. PMID- 15363744 TI - Disposition of abandoned embryos. PMID- 15363745 TI - Oocyte donation to postmenopausal women. PMID- 15363746 TI - Embryo splitting for infertility treatment. PMID- 15363747 TI - Use of fetal oocytes in assisted reproduction. PMID- 15363748 TI - Posthumous reproduction. PMID- 15363749 TI - Drug deaths in police custody: is dual diagnosis a significant factor? AB - This study examined cases for possible mental health issues based on a study of 43 deaths in custody that had been supervised by the PCA between 1998 and 2002 involving the use of drugs. In 18 of the 43 cases, there was evidence of one of three groups of mental health symptoms--in five cases, there was evidence of psychosis, in five of previous self-harm or suicidal attempts, and in a further eight, there were indications of anxiety or depression. This constitutes a total of 42% of the cases studied. Those with mental health factors were more likely to have swallowed the drugs used, were more likely to have used prescription drugs and were more likely to have been believed to be faking their symptoms by the officers involved in these cases. While it is recognised that mental health problems are widespread in the criminal justice system, the diversity of conditions and the marked overlap with the use of alcohol and illicit drugs has not been sufficiently recognised, either in the training of officers or in the procedures for intervention in the custody suite. PMID- 15363750 TI - Use of ApoB' hyper variable region in studying mixed chimerism and maternal contamination in North Indian populations. AB - ApoB3' hyper variable region is one of the highly polymorphic genetic marker and reveals a high degree of allelic variation in different populations therefore; it can be a useful marker for different clinical tests in which individual differences at DNA level form the basis of detection. In the present study we compared Apo B3 HVR with other 28 STR markers at allele frequency level, heterozygosity, polymorphism information content (PIC) and power of exclusion. Our results indicated a high degree of heterozygosity, PIC and power of exclusion for Apo B3 HVR. These criteria lead us to investigate this marker for different purposes like detection of maternal contamination in chorionic villus samples and chimerism studies after the engraftment of bone marrow in bone marrow transplantation patients. The utility of this marker has been discussed in comparison of other markers. PMID- 15363751 TI - A preliminary study identifying risk factors in drop-out from a prison therapeutic community. AB - Three different groups of therapy leavers and current residents (total n = 80) from Dovegate Prison's Therapeutic Community (TC) were compared on various demographic, criminogenic and psychometric attributes. Statistically significant findings distinguished the leaver groups from current residents and differentiated between type of leavers. Those leavers judged as unsuitable for therapy were characterised by high scores of Psychopathy, Neuroticism, Criminality, Addiction, Venturesomeness, and Avoidant and Schizoid Personality Disorders. Those leavers who had elected to leave the TC were more likely to have a sexual index offence, significant scores on the Multiphasic Sex Inventory, and Paranoid and Depressive Personality Disorders. Logistic Regression analysis confirmed that particular personality disorders were predictive of therapy leaver group membership. Paranoid Personality Disorder was identified as a predictor of all types of drop-out. These findings provide a psychological profile of the different types of residents who have left a prison therapeutic community and can be used to support and inform selection and assessment practices. PMID- 15363752 TI - The security of patient identifiable information in doctors' homes. AB - Ethically and legally doctors bear a responsibility to ensure the security of patient identifiable information in their possession. Many doctors, especially those in forensic medicine, hold paper or computerised medical records at home. This survey was conducted to assess the level of security for these records and awareness of the issues. Fifty-six forensic physicians (30 male, 26 female) answered a questionnaire. KEY FINDINGS: Eighty-nine percent used a computer to write patient notes and reports, but only 26 of these were on the Data Protection Register, and only 24 password-protected their files. Few doctors took steps to protect data on old computers they had stopped using. Of those responding, 88% held paper records at home but only of these had lockable filing cabinets. Burglar alarms were fitted in 77% of homes, yet 36% of homes had been burgled. No participants had written instructions for disposal of records and reports after their death. Older participants were more likely to have been burgled, yet less likely to have antiviral software than their younger counterparts. Participants expressed the need for information, education and training in data security. PMID- 15363753 TI - Ninhydrin-dyed latent fingerprints as a DNA source in a murder case. AB - In this case report, we describe the possibility of using ninhydrin-dyed fingerprints as a DNA source for STR typing. Preliminary tests prove that ninhydrin-dyed material still can be useful for STR typing. The case material consisted of seven ninhydrin-labeled latent fingerprints found at a murder crime site, which could not be typed in a classical manner. We were able to swap DNA from the ninhydrin-treated areas and successfully use it for STR typing. PMID- 15363754 TI - Fatal systemic allergic reaction following acrylic cranioplasty: a case report. AB - South Africa has a high death rate as a result of motor vehicle collisions. The motor vehicle accident fund is struggling with compensation claims made by the next of kin of the deceased. This report presents an unusual case of death as a result of systemic allergic reaction to acrylic material used to cover the frontal bone defect in a head injury patient. In independent forensic pathologist hired by the independent complaint directorate concluded that the cause of death was not related to the motor vehicle accident, and therefore the family cannot be compensated. The history, post mortem findings, management and issue of compensation claim are discussed. PMID- 15363755 TI - A syndrome of bilateral hemorrhage of the thalamus and myocarditis with fatal course. AB - We report the case of a 41-year-old patient with bilateral hemorrhage of the thalamus, leading to death. Post-mortem examination showed acute myocarditis. Neuropathological study showed perivascular infiltrates in affected thalamic regions. Laboratory investigation failed to find any causal agent. We hypothesize an infective agent, affecting the heart and thalamus, as the cause of this syndrome. Diaschisis due to the strategic anatomical position of the thalamus may have been responsible for coma state and death. PMID- 15363756 TI - Sudden unexpected death in an adult patient with endocardial cushion defect. AB - A 50-year-old male died following a road traffic accident. Postmortem examination revealed that the injuries caused by the accident did not seem to have caused his death. A large ostium primum defect was found, with bridging leaflets attached to the interventricular septum. In this article, we present a case of sudden death with partial endocardial cushion defect and discuss the mechanism of death. We believe that this is the first report of sudden death caused by endocardial cushion defect. PMID- 15363757 TI - Variants of homicide: a review. AB - Killing of an individual is the highest level of aggression found in all cultures. Modes of killing of the victims vary with the history and the cultural background of a country. In 13th century, pardonable homicide was distinguished from homicide committed with preplanning. Later homicide which was committed with preplanning was classified as 'premeditated' or 'intentional' or 'first degree' murder in different parts of the world. When sufficient proof of preplanning or malice aforethought was not present the term 'culpable homicide not amounting to murder' is used in India, 'manslaughter' in England and 'unintentional' homicide in Iceland. Sometime homicide may be the end result of several different chains of events of which mental abnormality is one. To convict a person in case of homicide the types of homicide must be established. This review explores some means of classifying homicides. PMID- 15363758 TI - The history of forensic podiatry. PMID- 15363759 TI - Forensic web watch--forensic nursing. AB - At times the boundaries between medicine and other allied health care professions becomes blurred such that roles historically undertaken by doctors are today undertaken by others often, in fact, more suited to the role. At the time of conception and implementation this exchange of duties may not be accepted by those trapped within the traditions of the hierarchy of medicine which often leads to conflict between the two groups who initially have to work side by side. A good example of the exchange of duties and the differences of forensic practice throughout the world is that of the role of the nurse in death and forensic investigation. This web watch will draw attention to sites dedicated to forensic nursing and highlight the roles which have been taken on by nurses, predominately within the United States, which have traditionally been viewed as the arena of the doctor. PMID- 15363760 TI - Prevalence of infectious diseases in feral cats in Northern Florida. AB - Objectives of this study were to determine prevalence of infection in feral cats in Northern Florida with a select group of infectious organisms and to determine risk factors for infection. Blood samples or sera from 553 cats were tested with a panel of antibody, antigen or PCR assays. Male cats were at higher risk for FIV, Mycoplasma haemofelis, and M. haemominutum. Infection with either FeLV or FIV was associated with increased risk for coinfection with the other retrovirus, M. haemofelis, or M. haemominutum. Bartonella henselae had the highest prevalence and was the only organism that did not have any associated risk for coinfection with other organisms. Feral cats in this study had similar or lower prevalence rates of infections than those published for pet cats in the United States. Thus, feral cats assessed in this study appear to be of no greater risk to human beings or other cats than pet cats. PMID- 15363761 TI - Anesthetic and physiologic effects of tiletamine, zolazepam, ketamine, and xylazine combination (TKX) in feral cats undergoing surgical sterilization. AB - Tiletamine (12.5 mg), zolazepam (12.5 mg), ketamine (20 mg), and xylazine (5 mg) (TKX; 0.25 ml, IM) combination was evaluated as an anesthetic in 22 male and 67 female adult feral cats undergoing sterilization at high-volume sterilization clinics. Cats were not intubated and breathed room air. Oxygen saturation (SpO(2)), mean blood pressure (MBP), heart rate (HR), respiration rate (RR), and core body temperature were recorded. Yohimbine (0.25 ml, 0.5 mg, IV) was administered at the completion of surgery. TKX produced rapid onset of lateral recumbency (4+/-1 min) and surgical anesthesia of sufficient duration to complete surgical procedures in 92% of cats. SpO(2) measured via a lingual pulse oximeter probe averaged 92+/-3% in male cats and 90+/-4% in females. SpO(2) fell below 90% at least once in most cats. MBP measured by oscillometry averaged 136+/-30 mm Hg in males and 113+/-29 mm Hg in females. MBP increased at the onset of surgical stimulation suggesting incomplete anti-nociceptive properties. HR averaged 156+/ 19 bpm, and RR averaged 18+/-8 bpm. Neither parameter varied between males and females or over time. Body temperature decreased significantly over time, declining to 38.0+/-0.8 degrees C at the time of reversal in males and 36.6+/-0.8 degrees C at the time of reversal in females. Time from anesthetic reversal to sternal recumbency was prolonged (72+/-42 min). Seven cats (8%) required an additional dose of TKX to maintain an adequate plane of anesthesia at the onset of surgery, and this was associated with significantly longer recovery times (108+/-24 min). PMID- 15363762 TI - Feline high-rise syndrome: 119 cases (1998-2001). AB - High-rise syndrome was diagnosed in 119 cats over a 4-year period. 59.6% of cats were younger than one year, and the average height of the fall was four stories. High-rise syndrome was more frequent during the warmer period of the year. 96.5% of the presented cats, survived after the fall. 46.2% of cats had fractured limbs; 38.5% of fractures were of the forelimb, 61.5% of the hindlimb. The tibia was fractured most often (36.4%), followed by the femur (23.6%). 78.6% of femoral fractures were distal. The mean age of patients with femoral fractures was 9.1 months, and with tibial fractures 29.2 months. Thoracic trauma was diagnosed in 33.6% of cats. Pneumothorax was diagnosed in 20% of cats, and pulmonary contusions in 13.4%. Falls from the seventh or higher stories, are associated with more severe injuries and with a higher incidence of thoracic trauma. PMID- 15363763 TI - Pain management in cats--past, present and future. Part 1. The cat is unique. AB - Cats are popular pets but until recently their perioperative and traumatic pain was seriously underestimated and under treated. There are several causes of this under treatment. First, it may be difficult to detect pain in cats, because they do not demonstrate overt pain-associated behaviour. Secondly, there are relatively few analgesic drugs with market authorization for cats. Thirdly, cats have an unfortunate reputation for toxicity from analgesic drugs, particularly opioid-induced mania and classical non steroidal anti inflammatory drug toxicity. Fourthly, cats are deficient in some metabolic pathways used to metabolise analgesic drugs in other species; this may lead to genuine toxicity or to lack of effect. Recently, understanding of feline behaviour and physiology has improved, leading to better clinical management of this enigmatic species. Behavioural methods are proving to be the best means of assessing pain, and knowledge of unique feline physiology has enabled rational treatment protocols to be developed specifically for cats. PMID- 15363764 TI - Pain management in cats--past, present and future. Part 2. Treatment of pain- clinical pharmacology. AB - Opioids have an unjustified reputation for causing mania in cats, but with refinements in dosing they are now used successfully in this species. The mu opioid agonists are generally considered the best analgesics. Morphine (0.1-0.3 mg/kg) is effective in a clinical setting. Methadone (up to 0.5 mg/kg) has a similar profile to morphine. Pethidine (Demerol, meperidine; 2-5 mg/kg) is a useful analgesic with a faster onset but shorter duration of action than morphine. Oxymorphone and hydromorphone (0.05-0.1 mg/kg) are widely used in the USA. These opioids are more potent (up to 10 times), and longer acting than morphine in cats. Butorphanol (0.1-0.4 mg/kg) is a mu-opioid antagonist that produces its analgesic actions through kappa agonist activity. It rapidly reaches a ceiling effect, is short acting and is a weaker analgesic than pure mu opioids. Buprenorphine (0.01-0.02 mg/kg), a partial mu-agonist, is the most popular opioid used in small animal practice in the UK, other parts of Europe, Australia and South Africa. In clinical studies it has produced better analgesia than several other opioids and appears to be highly suitable for perioperative pain management in cats. NSAIDs are also used in cats for pain management, although cats metabolise these differently from other species. With appropriate dosing, carprofen (1-4 mg/kg) and meloxicam (0.3 mg/kg) have proved highly effective with few side effects. The use of ketoprofen (2 mg/kg), tolfenamic acid (4 mg/kg) and vedaprofen (0.5 mg/kg) has been reported in cats. Other less traditional analgesics such as ketamine, medetomidine and local anaesthetics are also used for clinical pain management. The transmucosal, transdermal and epidural routes offer novel methods for administration of analgesic drugs and have considerable potential for improving techniques in feline pain management. PMID- 15363765 TI - Exhaled breath condensate (EBC) collection in cats--description of a non-invasive technique to investigate airway disease. AB - Exhaled breath condensate has been collected in other species and used as a non invasive method of evaluating airway disease by measurement of various markers in the fluid, including hydrogen peroxide, nitric oxide, leukotrienes and prostaglandins. We describe a novel technique for the collection of exhaled breath condensate from cats, which enabled collection of fluid and measurement of its hydrogen peroxide concentration. Further studies will be needed to establish the value of this technique in the investigation of feline respiratory disease. PMID- 15363766 TI - What is your diagnosis? PMID- 15363767 TI - Scheduling and preparation for endoscopic procedures: a biased perspective from a private practice. AB - Our ability to function successfully as efficient physicians depends substantially on our employees who surround us in our practices. It is our office associates who initially come into contact with our patients, generally spend more time with them than we do, and clearly play the major role in the scheduling of, and preparation for, endoscopic examinations. In this article we discuss ways in which we can improve scheduling and preparation for endoscopic procedures. PMID- 15363768 TI - Intraprocedural steps to enhance efficiency. AB - This section focuses on intraprocedural steps that can improve endoscopic efficiency. Contributors to procedural efficiency and inefficiency are considered. The limitations to enhancing efficiency are presented. Strategies for incorporating contributors to efficiency including techniques, technologies, accessories, and attitudes are presented. PMID- 15363769 TI - Efficiency issues in sedation and monitoring. AB - Sedation and monitoring are key elements of the endoscopy process. There continues to be intense study of better methods for sedation and monitoring to improve the endoscopic "experience" for both patient and physicians alike. Our current practices will likely change in the future with technologic advances (monitoring) and expansion of our pharmacologic armamentarium (sedation). PMID- 15363770 TI - Endoscopy unit form and function. AB - During the last 15 years, the demand for gastrointestinal endoscopic procedures has grown exponentially, and concurrently the endoscopic procedure unit has grown in structure and function. Within these units, much has already been done to maximize procedural efficiency and productivity while optimizing quality of care and patient satisfaction. Still, health care providers need to adapt and adjust continuously to the changing environment and seek novel ways for improving unit efficiency. This article addresses some concepts for efficiency in endoscopy unit structure and function. PMID- 15363771 TI - Lessons learned from the operating room about procedure efficiency. AB - The driving force for increasing efficiency in most endoscopy suites is to accommodate the flood of patients requiring screening colonoscopy. It is likely that in the next 5 to 10 years, another, less invasive screening test, will replace this demand. This article reviews some of the lessons learned from surgeon's and anesthesiologist's efforts to improve operating room (OR) efficiency and suggests ways to apply this information to running the endoscopy suite. PMID- 15363772 TI - Efficiency in endoscopy with improvements in instrumentation. AB - This article explores ways in which developments in endoscopic instrumentation might be used to improve efficiency. While this holds great promise, it is important to first define efficiency and make sure that the goals for improving efficiency through technology will actually benefit patients and physicians. PMID- 15363773 TI - Improving efficiency in endoscopy with robotic technology. AB - The adoption of robotic systems will alter endoscopy suite environments in terms of staff roles, work processes, and the physical layout and design of the workspace. PMID- 15363774 TI - Self-propelling endoscopes. AB - Self-propelling endoscopes offer exciting possibilities for improving access to colon cancer screening, safety of colonoscopy, and efficiency in endoscopy. From an operational perspective, efficiency in endoscopy may be increased or decreased by the introduction of a self-propelling endoscope, depending on the instrument's technical performance characteristics and capabilities, its safety profile, ease of use, the physician time required to review the endoscopic findings, and requirements for sedation, if any. In addition, patient acceptance of such new technology will be a driving force determining its potential for success in the competition for a niche in the diagnostic armamentarium of colon cancer screening. PMID- 15363775 TI - Smart endoscopes. AB - This article explores some of the innovations that already have been incorporated into endoscopes as well as imagines the kinds of advances that could transform endoscopes from passive tools into more active participants. It will be up to the planners and engineers and ultimately the marketplace to determine what is not only achievable but also useful. PMID- 15363776 TI - Endoscope reprocessing and repair costs. AB - Endoscope reprocessing and repair are important factors in regard to patient safety, optimal endoscope function, and costs. Endoscope reprocessing requires high level disinfection. There are three steps in reprocessing; mechanical cleaning, disinfection, and that is followed by rinsing and drying. The most critical step is mechanical cleaning which results in a 10 (4) reduction in microbial contamination or bioburden. Improper cleaning can overwhelm high-level disinfection regardless of subsequent steps and is the most susceptible to error. Endoscopy costs are dependent on costs of instruments, cleaning costs, and repair costs. The average repair costs at our endoscopy unit remained stable for more than 4 years suggesting a durable life of use. Consideration of endoscope accessories use should consider potential damage to endoscope use, cost to process re-useable items, sterility, and consideration of recent legislation on reprocessing of single-use devices. PMID- 15363777 TI - The impact of tissue sampling on endoscopy efficiency. AB - Much of the time spent in gastrointestinal endoscopy is devoted to tissue sampling. For this reason, efforts to facilitate tissue diagnosis can greatly influence the overall efficiency of endoscopy. This overview reviews the steps involved in tissue sampling and highlights potential ways to achieve increased efficiency in tissue sampling. Both potential improvements in process and innovations in technology are considered. PMID- 15363778 TI - Possible features of current electronic endoscopic information systems: what to look for. AB - The endoscopic electronic medical record (EEMR) has the potential to increase greatly the efficiency of both the endoscopist and the entire endoscopy unit. Potential efficiencies depend on the specific situation of the endoscopist and the type of endoscopy unit. This article describes a number of features available in current EEMRs as well as some emerging features that are important for future databases. Additionally, some aspects of equipment and staff needed to implement a successful EEMR are discussed. PMID- 15363779 TI - Electronic endoscopic information systems: what is out there? AB - This review summarizes some of the endoscopy electronic medical records (EEMRs) that are presently available. The objective is simply to familiarize the reader with some of the important systems and key features. It is not meant to be exhaustive, as a complete review of EEMRs would involve much more than a simple article; this document simply provides an introduction from which the groundwork can be laid. PMID- 15363780 TI - Toward more user-friendly electronic endoscopy information systems: role of accessories. AB - The needs placed on today's physician to improve efficiency in the endoscopy unit are nowhere more apparent than in transcribing endoscopy reports. In spite of the constant progress made in currently available electronic endoscopy information systems, the systems lack efficiency and ease of use. Both of these problems could be addressed through the linkage of electronic endoscopy information systems to currently available accessory systems. W review how the linkage of tablet personal computers, voice recognition systems, and web connectivity software, which facilitates communication with electronic-based medical records, can be helpful. The final answer, however, will have to await clinical trials. PMID- 15363783 TI - Partial biomimetic reconstitution of avian eggshell formation. AB - The avian eggshell is a biocomposite ceramic consisting of minute amounts of organic matrix and a crystalline calcium carbonate (calcite) filler. It is formed by a well regulated spatio-temporal assembling process, where extracellular matrix proteins, especially the sulfated glycosaminoglycan anionic sites of specific proteoglycans, have been involved in nucleation and growth of the inorganic crystalline phase. Together with such extracellular matrix molecules, the activity of carbonic anhydrase, is crucial for the normal eggshell formation. Here, we studied the effect of dermatan sulfate and carbonic anhydrase on the in vitro calcification of non-mineralized eggshell membrane-mammillae substrate at different pH and incubation times. Crystal morphology was analyzed by scanning electron microscopy. Crystal nucleation and growth was delayed at lower pH. Dermatan sulfate modified crystal morphology producing aggregates of large calcite crystals exhibiting a columnar morphology, contributing to the eggshell texture development. Carbonic anhydrase increased the velocity of crystal growth and eventually contributed to the fusion of the crystal aggregates to each other. Although, the effect of other macromolecules could not be ruled out, the combinatory effect of proteoglycans and carbonic anhydrase seems to be important for the control of eggshell formation. PMID- 15363784 TI - Use of X-ray computed microtomography for non-invasive determination of wood anatomical characteristics. AB - Quantitative analysis of wood anatomical characteristics is usually performed using classical microtomy yielding optical micrographs of stained thin sections. It is time-consuming to obtain high quality cross-sections from microtomy, and sections can be damaged. This approach, therefore, is often impractical for those who need quick acquisition of quantitative data on vessel characteristics in wood. This paper reports results of a novel approach using X-ray computed microtomography (microCT) for non-invasive determination of wood anatomy. As a case study, stem wood samples of a 2-year-old beech (Fagus sylvatica L.) and a 3 year-old oak (Quercus robur L.) tree were investigated with this technique, beech being a diffuse-porous and oak a ring-porous tree species. MicroCT allowed non invasive mapping of 2-D transverse cross-sections of both wood samples with micrometer resolution. Self-developed software 'microCTanalysis' was used for image processing of the 2-D cross-sections in order to automatically determine the inner vessel diameters, the transverse cross-sectional surface area of the vessels, the vessel density and the porosity with computer assistance. Performance of this new software was compared with manual analysis of the same micrographs. The automatically obtained results showed no significant statistical differences compared to the manual measurements. Visual inspection of the microCT slices revealed very good correspondence with the optical micrographs. Statistical analysis confirmed this observation in a more quantitative way, and it was, therefore, argued that anatomical analysis of optical micrographs can be readily substituted by automated use of microCT, and this without loss of accuracy. Furthermore, as an additional application of microCT, the 3-D renderings of the internal microstructure of the xylem vessels for both the beech and the oak sample could be reconstructed, clearly showing the complex nature of vessel networks. It can be concluded that the use of microCT in wood science offers an interesting potential for all those who need quantitative data of wood anatomical characteristics in either the 2-D or the 3-D space. PMID- 15363785 TI - Conformational epitope mapping of OmpC, a major cell surface antigen from Salmonella typhi. AB - The outer membrane protein OmpC, a trimer made of 16 stranded beta-barrel monomers, is a major cell surface antigen from the human pathogen Salmonella typhi. The relative stability of the epitopes recognising a Salmonella specific MAb (referred as MPN5) and an Enterobacteria specific MAb (referred as P7D8) and the role of the trimeric organisation has been probed using gel electrophoresis and monoclonal antibodies. The assembly of the trimer and the stability of the beta-barrel are found to be important for epitope presentation. The Salmonella specific conformational epitope is found to be more stable than the Enterobacteria specific one. The important residues of the Salmonella specific (Asp 25 of loop 1, Asp 340 of loop 8, Lys 334 of loop 8, and Tyr 210 of loop 5) and the Enterobacteria specific (Asp 25 of loop 1, Tyr 210 of loop 5, and Lys 152 of loop 4) conformational epitope have been identified using monoclonal antibodies, chemical modification, and solid phase binding methods. PMID- 15363786 TI - Structure of DNA sequence d-TGATCA by two-dimensional nuclear magnetic resonance spectroscopy and restrained molecular dynamics. AB - The 5' d-TpG 3' element is a part of DNA sequences involved in regulation of gene expression and is also a site for intercalation of several anticancer drugs. Solution conformation of DNA duplex d-TGATCA containing this element has been investigated by two-dimensional NMR spectroscopy. Using a total of 12 torsional angles and 121 distance constraints, structural refinement has been carried out by restrained molecular dynamics (rMDs) in vacuum up to 100 ps. The structure is characterized by a large positive roll at TpG/CpA base pair step and large negative propeller twist for AT and TA base pairs. The backbone torsional angle, gamma(O5'-C5'-C4'-C3'), of T1 residue adopts a trans-conformation which is corroborated by short intra nucleotide T1H6-T1H5' (3.7A) distance in nuclear overhauser effect spectroscopy (NOESY) spectra while the backbone torsional angle, beta(P-O5'-C5'-C4'), exists in trans as well as gauche state for T1 and C5 residues. There is evidence of significant flexibility of the sugar-phosphate backbone with rapid inter-conversion between two different conformers at TpG/CpA base pair step. The base sequence dependent variations and local structural heterogeneity have important implications in specific recognition of DNA by ligands. PMID- 15363787 TI - Sequence context and modified hydrophobic moment plots help identify 'horizontal' surface helices in transmembrane protein structure prediction. AB - Transmembrane proteins make up at least one-fifth of the genome of most organisms and are critical components of key pathways for cell survival and interactions with the environment. The function of helices found at the membrane surface in transmembrane proteins has not been greatly explored, but it is likely that they play an ancillary role to membrane spanning helices and are analogous to the surface active helices of peripheral membrane proteins, being involved in: lipid association, membrane perturbation, transmembrane signal transduction and regulation, and transmembrane helical bundle formation. Due to the difficulties in obtaining high-resolution structural data for this class of proteins, structure-from-sequence predictive methods continue to be developed as a means to obtain structural models for these largely intractable systems. A simple but effective variant of the hydrophobic moment analysis of amino acid sequences is described here as part of a protocol for distinguishing helical sequences that are parallel to or 'horizontal' at the membrane bilayer/aqueous phase interface from helices that are membrane-embedded or located in extra-membranous domains. This protocol when tested on transmembrane spanning protein amino acid sequences not used in its development, was found to be 84-91% accurate when the results were compared to the partition locations in the corresponding structures determined by X-ray crystallography, and 72% accurate in determining which helices lie horizontal or near horizontal at the lipid interface. PMID- 15363788 TI - The influence of DNA stiffness upon nucleosome formation. AB - The rotational and translational positioning of nucleosomes on DNA is dependent to a significant extent on the physicochemical properties of the double helix. We have investigated the influence of the axial flexibility of the molecule on the affinity for the histone octamer by substituting selected DNA sequences with either inosine for guanosine or diaminopurine for adenine. These substitutions, respectively, remove or add a purine 2-amino group exposed in the minor groove and, respectively, decrease and increase the apparent persistence length. We observe that for all sequences tested inosine substitution, with one exception, increases the affinity for histone binding. Conversely diaminopurine substitution decreases the affinity. In the sole example where replacement of guanosine with inosine decreases the persistence length as well as the affinity for histones, the substitution concomitantly removes an intrinsic curvature of the DNA molecule. We show that, to a first approximation, the binding energy of DNA to histones at 1M NaCl is directly proportional to the persistence length. The data also indicate that a high local flexibility of DNA can favour strong rotational positioning. PMID- 15363789 TI - Crystal structure of a novel antifungal protein distinct with five disulfide bridges from Eucommia ulmoides Oliver at an atomic resolution. AB - EAFP2 is a novel antifungal protein isolated from the bark of the tree Eucommia ulmoides Oliver. It consists of 41 residues and is characterized with a five disulfide motif and the inhibitory effects on the growth of both cell wall chitin containing and chitin-free fungi. The crystal structure of EAFP2 at an atomic resolution of 0.84 A has been determined by using Shake-and-Bake direct methods with the program SnB. The phases obtained were of sufficient quality to permit the initial model built automatically and the structural refinement carried out using anisotropic displacement parameters resulted in a final crystallographic R factor of 6.8%. In the resulting structural model, all non-hydrogen protein atoms including an unusual pyroglutamyl acid residue at the N-terminal can fit to the articulated electron densities with one centre and more than 65% of the hydrogen atoms in the protein can be observed as individual peaks in the difference map. The general fold of EAFP2 is composed of a 3(10) helix (Cys3-Arg6), an alpha helix (Ala27-Cys31) and a three-stranded antiparallel beta-sheet (Cys16-Ser18, Cys23-Ser25, and Cys35-Cys37) and cross-linked by five disulfide bridges. The tertiary structure of EAFP2 can be divided into two structural sectors, A and B. Sector A composed of residues 11-30 adopts a conformation similar to the chitin binding domain in the hevein-like proteins and features a hydrophobic surface embraced a chitin-binding site (Tyr20, 22, 29, and Ser18). The distinct disulfide bridge Cys7-Cys37 connects the N-terminal ten residues with the C-terminal segment 35-41 to form the sector B, which features a cationic surface distributing all four positively charged residues, Arg6, 9, 36, and 40. Based on these structural features, the possible structural basis of the functional properties of EAFP2 is discussed. PMID- 15363790 TI - Crystal structure of Bacillus subtilis YckF: structural and functional evolution. AB - The crystal structure of the YckF protein from Bacillus subtilis was determined with MAD phasing and refined at 1.95A resolution. YckF forms a tight tetramer both in crystals and in solution. Conservation of such oligomerization in other phosphate sugar isomerases indicates that the crystallographically observed tetramer is physiologically relevant. The structure of YckF was compared to with its ortholog from Methanococcus jannaschii, MJ1247. Both of these proteins have phosphate hexulose isomerase activity, although neither of the organisms can utilize methane or methanol as source of energy and/or carbon. Extensive sequence and structural similarities with MJ1247 and with the isomerase domain of glucosamine-6-phosphate synthase from Escherichia coli allowed us to group residues contributing to substrate binding or catalysis. Few notable differences among these structures suggest possible cooperativity of the four active sites of the tetramer. Phylogenetic relationships between obligatory and facultative methylotrophs along with B. subtilis and E. coli provide clues about the possible evolution of genes as they loose their physiological importance. PMID- 15363791 TI - Mapping the anatomy of a Plasmodium falciparum MSP-1 epitope using pseudopeptide induced mono- and polyclonal antibodies and CD and NMR conformation analysis. AB - Antigen structure modulation represents an approach towards designing subunit malaria vaccines. A specific epitope's alpha carbon stereochemistry, as well as its backbone topochemistry, was assessed for obtaining novel malarial immunogens. A variety of MSP-1(38-61) Plasmodium falciparum epitope pseudopeptides derived were synthesised, based on solid-phase pseudopeptide chemistry strategies; these included all-L, all-D, partially-D substituted, all-Psi-[NH-CO]-Retro, all-Psi [NH-CO]-Retro-inverso, and Psi-[CH2NH] reduced amide surrogates. We demonstrate that specific recombinant MSP-1(34-469) fragment binding to red blood cells (RBCs) is specifically inhibited by non-modified MSP-1(42-61), as well as by its V52-L53, M51-V52 reduced amide surrogates and partial-D substitutions in K48 and E49. In vivo tests revealed that reduced amide pseudopeptide-immunised Aotus monkeys induced neutralising antibodies specifically recognising the MSP-1 N terminus region. These findings support the role of molecular conformation in malaria vaccine development. PMID- 15363792 TI - The localization of occluded matrix proteins in calcareous spicules of sea urchin larvae. AB - The sea urchin embryo forms calcareous endoskeletal spicules composed of calcite and an occluded protein matrix. Though the latter is approximately 0.1% of of the mass, the composite has substantially altered material properties, e.g., conchoidal fracture planes and increased hardness. Experiments were conducted to examine the localization of matrix proteins occluded in the mineral by use of immunocytochemistry coupled with scanning electron microscopy (SEM). The isolated, unfixed spicules were etched under relatively gentle conditions and exposed to affinity purified antibodies made against two different matrix proteins, as well as an antibody to the entire constellation of matrix proteins. Immunogold tagged secondary antibody was used to observe antibody localization in the back scatter mode of SEM. All proteins examined were very widely distributed throughout the calcite, supporting a model of the structure in which a multiprotein assemblage is woven with fine texture around microcrystalline domains of calcite. Gentle etching revealed a laminar arrangement of calcite solubility, consistent with a stepwise deposition of matrix and mineral to increase girth of the spicule. PMID- 15363793 TI - Cryo-electron microscopy of vitreous sections of native biological cells and tissues. AB - Cryo-electron microscopy of vitreous sections (CEMOVIS) is, in principle, the ultimate method of specimen preparation. It consists in ultra-rapid cooling of a sizable sample of biological material that is cut into thin sections. These are subsequently observed at low temperature in their fully hydrated vitreous state. Here, we show that CEMOVIS reveals the native state of cells and tissues with unprecedented quality and resolution. What is seen differs considerably from what conventional electron microscopy has shown previously and it is seen with more details. Our findings are demonstrated with images of cyanobacteria and skin. PMID- 15363795 TI - A prehistory of cell adhesion. AB - Cell adhesion is a basic property of animal cells, but is also present in many other eukaryotes. Did cell adhesion systems arise independently in different eukaryotic groups, or do they share common origins? Recent results show that cell adhesion proteins related to cadherin, IgG-like CAM and C-type lectin are present both in sponges, the most distant animal branch, and in eukaryote groups outside the metazoan lineage, indicating that these forms of adhesion arose prior to animal evolution. Furthermore, proteins containing features of animal adhesion systems, such as Fas-1 and thrombospondin domains, are distributed throughout the eukaryotes and function in cell adhesion. PMID- 15363796 TI - SED1 function during mammalian sperm-egg adhesion. AB - A prerequisite for successful fertilization is the species-specific binding of sperm to the extracellular coat of the egg. Gamete binding triggers the release of sperm hydrolytic enzymes that digest a path through the egg coat, thus bringing sperm into proximity with the egg plasma membrane where gamete fusion occurs. Although some components of the sperm membrane and the egg coat that participate in sperm-egg interactions have been identified, results from targeted deletions and gene substitutions indicate that other, as yet unidentified, gamete receptors must contribute to sperm-egg binding. Recent studies implicate the bi motif protein, SED1, as being required for successful sperm-egg adhesion in mouse. SED1 contains Notch-like EGF repeats as well as discoidin/F5/8 complement domains--motifs that mediate a variety of cell-cell and cell-matrix interactions. SED1's ability to promote gamete adhesion resides within its two discoidin/F5/8C domains, which are able to dock to substrates as diverse as phospholipid membranes and extracellular matrices. SED1 is also expressed in a wide range of tissues and epithelia, where it may function similarly as an adhesive protein facilitating cell-cell and/or cell-matrix interactions. PMID- 15363797 TI - Skin tight: cell adhesion in the epidermis of Caenorhabditis elegans. AB - The powerful genetics, genomics and microscopy tools available for C. elegans make it well suited to studying how epithelial cells adhere to one another and the extracellular matrix, and how the integrated, simultaneous activities of multiple cell adhesion complexes function to shape an organism. Recent studies using forward and reverse genetics have shed light on how phylogenetically conserved cell adhesion complexes, such as the cadherin/catenin complex, claudins, the Discs large complex and hemidesmosome-like attachment structures, regulate epithelial cell adhesion, providing new insights into conserved cell adhesion mechanisms in higher eukaryotes. PMID- 15363798 TI - A junctional problem of apical proportions: epithelial tube-size control by septate junctions in the Drosophila tracheal system. AB - The size of epithelial tubes is critical for the function of organs such as the lung, kidney and vascular system. However, the molecular mechanisms regulating tube size are largely unknown. Recent work in the Drosophila tracheal system reveals that septate junctions play a previously unsuspected role in tube-size control. Surprisingly, this tube-size function is distinct from the established diffusion barrier function of septate junctions, and involves regulation of cell shape rather than cell number. Possible tube-size functions of septate junctions include patterning of the apical extracellular matrix and regulation of conserved cell polarity genes such as Scribble and Discs Large. PMID- 15363799 TI - Plasmodesmata form and function. AB - Intercellular transport via plasmodesmata controls cell fate decisions in plants, and is of fundamental importance in viral movement, disease resistance, and the spread of RNAi signals. Although plasmodesmata appear to be unique to plant cells, they may have structural and functional similarities to the newly discovered tunneling nanotubes that connect animal cells. Recently, proteins that localize to plasmodesmata have been identified, and a microtubule-associated protein was found to negatively regulate the trafficking of viral movement proteins. Other advances have delivered new insights into the function and molecular nature of plasmodesmata and have shown that protein trafficking through plasmodesmata is developmentally regulated, opening up the possibility that the genetic control of plasmodesmal function will soon be understood. PMID- 15363800 TI - Connexins: functions without junctions. AB - Emerging studies indicate that connexins have activities completely unrelated to gap junctions and, conversely, that non-connexin proteins can form gap junction channels. PMID- 15363801 TI - Biology and pathology of nectins and nectin-like molecules. AB - Immunoglobulin-like nectins contribute to the formation of a variety of cell-cell junctions, acting cooperatively with, or independently of, cadherins. In addition, nectins heterophilically trans-interact with nectin-like molecules (Necls), which are involved in cell adhesion, migration, and proliferation, and assist or modify their functions. On the other hand, nectins and Necls serve as viral receptors and are associated with human diseases (including cancer) when mutated or upregulated. PMID- 15363802 TI - Protecting your tail: regulation of cadherin degradation by p120-catenin. AB - Work in various model systems has yielded conflicting views of how p120-catenin participates in adherens junction assembly and regulation. A series of recent studies indicate that a core function of p120-catenin in mammalian cells is to regulate cadherin turnover by modulating the entry of cadherins into degradative endocytic pathways. By this mechanism, cellular levels of p120-catenin perform a 'rheostat' or 'set point' function that controls steady-state cadherin levels. These studies parallel a growing interest in the regulation of cadherin levels at the cell surface by membrane trafficking pathways. Collectively, the findings suggest exciting new roles for p120-catenin at the interface between cadherins and membrane trafficking machinery, and imply novel mechanisms by which p120 catenin may regulate cell adhesion and migration in the context of development and cancer. PMID- 15363803 TI - Adenomatous polyposis coli proteins and cell adhesion. AB - Adenomatous polyposis coli (APC) is an important tumour suppressor in the mammalian intestinal epithelium. It binds to beta-catenin and its role as a tumour suppressor depends predominantly on its ability to downregulate soluble beta-catenin, a key effector of the Wnt signalling pathway. However, epithelial cells have a distinct subcellular pool of beta-catenin, or Drosophila Armadillo, which functions as a structural component of adherens junctions. Notably, APC proteins can be associated with these adherens junctions, and recent evidence points to a role for APC in cellular adhesion. Thus, APC--like beta catenin/Armadillo--may have a dual role in Wnt signal transduction and in cellular adhesion, which could be relevant to its activity as a tumour suppressor. PMID- 15363804 TI - Desmosomes and disease: pemphigus and bullous impetigo. AB - Desmosomal cadherins are the pathophysiologic targets of autoimmune or toxin mediated disruption in the human diseases pemphigus and bullous impetigo (including its generalized form, called staphylococcal scalded skin syndrome). Experiments exploiting the production of both pathogenic and nonpathogenic antidesmoglein antibodies in pemphigus patients' sera have afforded data that make an invaluable contribution towards identifying the functional domains of the desmogleins involved in intercellular adhesion. Conformational epitopes of antidesmoglein autoantibodies in pemphigus patients' sera and the specific cleavage site of desmoglein 1 by exfoliative toxin have been identified, implicating the N-terminal extracellular domains of the desmogleins as critical regions for controlling intercellular adhesion. Furthermore, the development of active autoimmune mouse models for pemphigus allows in vivo characterization of the disease and its pathogenesis. These studies offer new insight into the potential mechanisms of acantholysis in pemphigus and staphylococcal-associated blistering disease, with implications for the role of desmogleins in desmosomal structure and function. PMID- 15363805 TI - Regulation of integrin function through conformational complexity: not simply a knee-jerk reaction? AB - Such diverse biological processes as the maintenance of tissue architecture and the regulation of cell migration are controlled through dynamic changes in integrin receptor conformation. Early analyses of the mechanisms of shape change by integrins led to the definition of three inter-convertible conformational states: inactive, primed and ligand-occupied. Recent advances reviewed in this article have now shown that the integrin molecule contains a number of flexible joints and connections, leading to a broad spectrum of possible conformational states. This conformational complexity is likely to permit fine-tuning of integrin function through regulation of ligand-binding affinity and intracellular signalling. PMID- 15363806 TI - Roles of alphav integrins in vascular biology and pulmonary pathology. AB - The five integrins that contain the alphav subunit are widely expressed and their expression is tightly regulated. However, most tissues in mice lacking the alphav subunit, and thus deficient in all five integrins, develop normally, suggesting that nearly all of the critical steps in development and cellular differentiation can occur in the absence of these integrins. Studies over the past few years have identified highly specialized roles for specific alphav integrins in preventing inappropriate vascular growth and in control of vascular permeability. Two members of this family, alphavbeta6 and alphavbeta8, play novel roles in activating latent complexes of the growth factor TGFbeta (transforming growth factor beta). Studies in mice lacking the beta6 subunit have identified unexpected roles for alphavbeta6-mediated TGFbeta activation in models of pulmonary and renal fibrosis, acute lung injury and pulmonary emphysema. PMID- 15363807 TI - Regulation of matrix biology by matrix metalloproteinases. AB - Matrix metalloproteinases (MMPs) are endopeptidases that contribute to growth, development and wound healing as well as to pathologies such as arthritis and cancer. Until recently, it has been thought that MMPs participate in these processes simply by degrading extracellular matrix (ECM) molecules. However, it is now clear that MMP activity is much more directed and causes the release of cryptic information from the ECM. By precisely cleaving large insoluble ECM components and ECM-associated molecules, MMPs liberate bioactive fragments and growth factors and change ECM architecture, all of which influence cellular behavior. Thus, MMPs have become a focal point for understanding matrix biology. PMID- 15363808 TI - Integrin-linked kinase: integrin's mysterious partner. AB - Integrin-mediated cell adhesion regulates a vast number of biological processes including migration, survival and proliferation of cells. It is therefore not surprising that defects in integrin function are often rate-limiting for development and profoundly affect the progression of several diseases. The functions of integrins are mediated through the recruitment of cytoplasmic plaque proteins. One of these is integrin-linked kinase, which connects integrins to the actin cytoskeleton and transduces signals through integrins to the extracellular matrix and from integrins to various subcellular compartments. PMID- 15363809 TI - Assembly and tissue functions of early embryonic laminins and netrins. AB - Vertebrate laminins and netrins share N-terminal domain structure, but appear to be only distantly related. Both families can be divided into different subfamilies on the basis of structural considerations. Recent observations suggest that specific laminin and netrin members have developmental functions that are highly conserved across species. Vertebrate laminin-1 (alpha1beta1gamma1) and laminin-10 (alpha5beta1gamma1), like the two Caenorhabditis elegans laminins, are embryonically expressed and are essential for basement membrane assembly. Basement membrane assembly is a cooperative process in which laminins polymerize through their LN domains and anchor to the cell surface through their G domains; this leads to cell signaling through integrins and dystroglycan (and possibly other receptors) recruited to the adherent laminin. Netrins may associate with this network through heterotypic LN domain interactions. Vertebrate netrin-1, like invertebrate UNC-6/netrins, is well known as an extracellular guidance cue that directs axon migration towards or away from the ventral midline. It also regulates cell adhesions and migrations, probably as a basement membrane component. Although sharing structural features, these two vertebrate protein families are quite distinct, having both retained members that mediate the ancestral developmental functions. PMID- 15363810 TI - Eph/ephrin signaling in morphogenesis, neural development and plasticity. AB - Ephrins are cell-surface-tethered ligands for Eph receptors, the largest family of receptor tyrosine kinases. During development, the Eph/ephrin cell communication system appears to influence cell behavior such as attraction/repulsion, adhesion/de-adhesion and migration, thereby influencing cell fate, morphogenesis and organogenesis. During adulthood, the Eph/ephrin system continues to play roles in tissue plasticity, for example in shaping dendritic spines during neuronal plasticity. Mechanistically, Eph-ephrin repulsive behavior appears to require ligand-receptor internalization and signaling to Rho GTPases. PMID- 15363811 TI - Roles of fascin in cell adhesion and motility. AB - Many cell interactions depend on the assembly of cell protrusions; these include cell attachment and migration in the extracellular matrix, cell-cell communication, and the ability of cells to sense their local environment. Cell protrusions are extensions of the plasma membrane that are supported internally by actin-based structures that impart mechanical stiffness. Fascin is a small, globular actin-bundling protein that has emerging roles in diverse forms of cell protrusions and in cytoplasmic actin bundles. The fascin-actin interaction is under complex regulation from the extracellular matrix, peptide factors and other actin-binding proteins. Recent developments advance our understanding of the multifaceted regulation of fascin and the roles of fascin-containing structures in cell adhesion, motility and invasion in the life of vertebrate organisms. PMID- 15363812 TI - Amlodipine increases nitric oxide production in exhaled air during exercise in patients with essential hypertension. AB - BACKGROUND: Endothelial production of nitric oxide (NO) is attenuated in patients with essential hypertension. We investigated whether treatment with amlodipine increased exhaled NO output (VNO) at rest and during exercise in patients with essential hypertension. METHODS: We studied the effect of amlodipine in seven untreated hypertensive patients. Cardiopulmonary exercise testing and NO measurement of exhaled air were performed on these patients before and after 2 months of amlodipine treatment. RESULTS: Amlodipine decreased blood pressure (BP) both at rest and during exercise (at rest: 147.1 +/- 6.4 [SEM]/89.9 +/- 4.4 v 133.6 +/- 5.4/82.7 +/- 3.9 mm Hg, P <.05; at peak exercise: 224.9 +/- 8.0/113.1 +/- 5.3 v 207.0 +/- 6.0/100.7 +/- 5.0 mm Hg, P <.05) without affecting heart rate (at rest: 67.6 +/- 3.9 v 70.4 +/- 4.5 beats/min, P =.33; peak exercise: 146.4 +/- 7.4 v 144.0 +/- 7.2 beats/min, P =.49). Amlodipine did not affect minute ventilation (VE) at rest or during exercise. It did not alter anaerobic threshold, peak oxygen uptake (peak VO(2)), or peak workload. However, after amlodipine treatment, VNO was significantly greater both at rest (130.8 +/- 19.4 v 180.4 +/- 24.8 nL/min, P <.05) and at peak exercise (380.0 +/- 47.5 v 582.6 +/- 74.3 nL/min, P <.05). CONCLUSIONS: Amlodipine increased NO production, at least in the pulmonary circulation, in patients with essential hypertension. In addition to its antihypertensive effect, the enhancement of NO production by amlodipine in the vasculature of other organs may contribute to its beneficial effects on the cardiovascular system. PMID- 15363813 TI - Antihypertensive efficacy of night-time graded-release diltiazem versus morning amlodipine in African Americans. AB - BACKGROUND: The effect of a once daily night-time (10 pm) graded-release diltiazem (GRD) on early morning blood pressure (BP), heart rate (HR), and rate pressure product (RPP) were compared with the effect of morning (8 am) amlodipine in 262 African American individuals with hypertension. METHODS: The multicenter, randomized, double-blind, parallel-group, dose-to-effect trial evaluated changes from baseline in BP, HR, and RPP (HR x systolic BP) by ambulatory BP monitoring during the first 4 h after awakening (diastolic BP = primary), between 6 am and 12 noon, and over a 24-h period. Patients were randomized to night-time GRD 360 mg (n = 132) or morning amlodipine 5 mg (n = 130) for 6 weeks, and were titrated to GRD 540 mg or amlodipine 10 mg after 6 weeks if clinic systolic BP/diastolic BP (SBP/DBP) was > or = 130/85 mm Hg. RESULTS: Compared with amlodipine, GRD showed significantly greater DBP reductions of 3.5 mm Hg (P < .0049) and 3.2 mm Hg (P < .0019) during the first 4 h after awakening and between 6 am and 12 noon respectively, as well as comparable reduction for the 24-h mean DBP. The SBP reductions during the morning periods were comparable, but the reduction in the 24-h mean SBP was 3.4 mm Hg greater (P < .0022) for amlodipine. Mean reductions in HR and RPP were significantly greater (P < or = .0008) for GRD during all intervals; amlodipine increased whereas diltiazem reduced HR with mean differences of 6.7 to 9.3 beats/min. Both treatments were well tolerated. CONCLUSIONS: Night-time GRD was more effective than morning amlodipine in reducing early morning DBP, HR, and RPP, as well as 24-h HR and RPP in African American individuals with hypertension. Amlodipine was more effective in reducing SBP over the 24-h period. PMID- 15363814 TI - Amlodipine attenuates oxidative stress-induced hypertension. AB - BACKGROUND: Dihydropyridine Ca2+-blockers, frequently used as antihypertensive and antianginal agents, have been found to exert potent antioxidant and cytoprotective activities against free radical-mediated vascular injury. METHODS: In the current study we examined the effect of amlodipine (AMLOD) on oxidative stress-induced hypertension in Sprague-Dawley rats administered buthionine sulfoximine (BSO), a glutathione (GSH) synthase inhibitor, in the drinking water. The control animals received drug-free water. Blood pressure (BP) was measured by tail-cuff plethysmography. Plasma levels of total 8-isoprostane, thromboxane A2, prostacyclin, nitric oxide, and aortic cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were determined by enzyme immunoassay. Plasma, kidney, and heart GSH were analyzed by high-performance liquid chromatography. RESULTS: Administration of BSO significantly increased BP, isoprostane, and thromboxane A2, whereas GSH, PGI2, and cAMP were reduced. When given alone, AMLOD alone reduced BP and the plasma levels of isoprostane and thromboxane A2, and elevated prostacyclin, nitric oxide, cGMP, and cAMP. When administered with BSO, AMLOD reversed the BSO-induced elevation of BP, isoprostane, and thromboxane A2 as well as the reduction in prostacyclin, cAMP, and cardiac GSH levels. CONCLUSIONS: The antihypertensive effect of amlodipine involves a reduction in oxidative stress, which appears to be mediated in part by the prostanoid endothelium-derived factors and nitric oxide. PMID- 15363815 TI - Beneficial and deleterious effects of rosiglitazone on hypertension development in spontaneously hypertensive rats. AB - The antihypertensive effect of the peroxisome proliferator-activated receptor (PPAR)gamma agonist rosiglitazone has been reported in patients with diabetes or obesity. The correlation of PPARgamma expression with blood pressure and the therapeutic application of rosiglitazone in spontaneously hypertensive rats (SHR) were investigated in the present study. Systolic blood pressure of 21-week SHR was significantly higher than that of age-matched Wistar-Kyoto rats (WKY) (225 +/ 5 v 144 +/- 2 mm Hg, P <.05). Basal expression levels of PPARgamma proteins in vascular tissues of 21-week SHR were significantly lower than that of age-matched 21-week WKY (P <.05). This reduced expression of PPARgamma was not detected between 5- and 13- week SHR and age-matched WKY. Cardiac PPARgamma expression was also not different among different age groups between SHR and WKY. Chronic treatment with rosiglitazone, but not PPARalpha agonist Wy14643, significantly retarded hypertension development and reversed abnormally faster heart rate in young SHR. An unfavorable effect of rosiglitazone treatment was the increased heart-to-body weight ratio accompanied by left ventricular hypertrophy. In conclusion, vascular PPARgamma protein expression in adult SHR (21 weeks) is significantly decreased in comparison with the age-matched WKY. Chronic rosiglitazone treatment retards hypertension development, but the associated prohypertrophy effect calls for a cautious use of this thiazolidinedindione in the treatment of insulin resistance syndrome associated with hypertension. PMID- 15363816 TI - Effect of eprosartan on cytoplasmic free calcium mobilization, platelet activation, and microparticle formation in hypertension. AB - BACKGROUND: Hypertensive patients show greater platelet activation than do normotensive individuals. Platelet activation is characterized by increased phosphatidylserine (PS) exposure in the external hemilayer of the membrane, a larger number of platelet microparticles (PMP), and changes in intraplatelet-free calcium kinetics. This study evaluated whether eprosartan can protect against undesirable platelet activation. METHODS: A total of 30 hypertensive patients (systolic blood pressure [SBP] 140 to 189 mm Hg; diastolic blood pressure [DBP] 90 to 109 mm Hg) without renal, liver, or cardiac organic lesions and with a mean age of 47.6 +/- 9.4 years and mean body mass index (BMI) of 27.9 +/- 3.9 kg/m2 received eprosartan (600 mg/day). They were compared with 31 normotensive individuals with a mean age of 43.3 +/- 6.7 years and a mean BMI of 26.8 +/- 3.9 kg/m2. Blood pressure measurements and platelet function changes were assessed at baseline (control and hypertensive patients) and after 1 and 2 months of eprosartan monotherapy (hypertensive patients only). RESULTS: Significant baseline to endpoint (month 2) changes in SBP and DBP were noted in the eprosartan group (SBP: baseline 152.2 +/- 16.8 mm Hg, endpoint 142.2 +/- 16.9 mm Hg, P <.01; DBP: baseline 93.5 +/- 9.9 mm Hg, endpoint 85.8 +/- 11.9 mm Hg, P <.001). Native circulating activated platelets increased in both groups after shear stress or Ca2+ ionophore activation, and were reduced by eprosartan (after shear exposure from 104% at month 1 to 76% after 2 months of therapy). Eprosartan therapy normalized the number of microparticles after blood shear exposure (P <.01) and after exposure to Ca2+ ionophore activation (P <.05) and significantly reduced the trend for platelets to be more readily activated in hypertensive compared with normotensive subjects (baseline to endpoint change P <.001; increase/shear versus baseline P <.001). Eprosartan partially normalizes cytoplasmic-free calcium mobilization in platelets. CONCLUSIONS: Eprosartan significantly reduces blood pressure and normalizes undesirable changes in platelet function. PMID- 15363817 TI - Matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in hypertension and their relationship to cardiovascular risk and treatment: a substudy of the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT). AB - BACKGROUND: Hypertension results in structural changes to the cardiac and vascular extracellular matrix (ECM). Matrix metalloproteinases (MMP) and their inhibitors (TIMP) may play a central role in the modulation of this matrix. We hypothesized that both MMP-9 and TIMP-1 would be abnormal in hypertension, reflecting alterations in ECM turnover, and that their circulating levels should be linked to cardiovascular (CHD) and stroke (CVA) risk scores using the Framingham equation. Second, we hypothesized that treatment would result in changes in ECM indices. METHODS: Plasma MMP-9 and TIMP-1 were measured before and after treatment (median 3 years) from 96 patients with uncontrolled hypertension participating in the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT). Pretreatment values were compared to circulating MMP-9 and TIMP-1 levels in 45 age- and sex-matched healthy controls. RESULTS: Circulating pretreatment MMP-9 and TIMP-1 levels were significantly higher in patients with hypertension than in the normotensive controls (P =.0041 and P =.0166, respectively). Plasma MMP-9 levels decreased, and TIMP-1 levels increased after treatment (P =.035 and P =.005, respectively). Levels of MMP-9 correlated with CHD risk (r = 0.317, P =.007) and HDL cholesterol (r = -0.237, P =.022), but not CVA risk. There were no significant correlations between TIMP-1 and CVA or CHD scores. CONCLUSIONS: Increased circulating MMP-9 and TIMP-1 at baseline in patients with hypertension could reflect an increased deposition and retention of type I collagen at the expense of other components of ECM within the cardiac and vascular ECM. After cardiovascular risk management, MMP-9 levels decreased and TIMP-1 levels increased. Elevated levels of MMP-9 also appeared to be associated with higher Framingham cardiovascular risk scores. Our observations suggest a possible role for these surrogate markers of tissue ECM composition and the prognosis of cardiovascular events in hypertension. PMID- 15363818 TI - Tissue inhibitor of metalloproteinase-1 and matrix metalloproteinase-9 levels in patients with hypertension Relationship to tissue Doppler indices of diastolic relaxation. AB - BACKGROUND: Hypertension, hypertensive heart disease, and left ventricular (LV) hypertrophy are integral to symptomatic diastolic heart failure. Tissue inhibitor of metalloproteinase-1 (TIMP-1) is linked to extracellular matrix fibrosis and is elevated in hypertension. We hypothesized a link between circulating TIMP-1, matrix metalloproteinase-9 (MMP-9), and resting echocardiographic LV filling parameters using tissue Doppler parameters of diastolic (dys)function. METHODS: Circulating MMP-9 and TIMP-1 levels were measured in citrated plasma by ELISA in 74 patients with hypertension (58 men, mean age 58 +/- 11 years) and 34 controls (23 men, mean age 53 +/- 13 years). All had confirmed normal short axis systolic contractility, with no significant wall motion abnormalities; the LV mass and standard resting tissue Doppler echocardiographic indices of diastolic function were also recorded. RESULTS: Both MMP-9 and TIMP-1 levels were higher in the hypertensive group (P =.0039 and P =.0054, respectively). When compared to controls, hypertensive patients had a greater LV mass (P =.0054), and differences in many of the parameters reflecting diastolic dysfunction (controls versus hypertensives: E: 0.71 +/- 0.15 v 0.81 +/- 0.15 m/sec, P =.004; A: 0.66 +/- 0.12 v 0.81 +/- 0.16 m/sec, P <.0001; e': 0.12 (0.09-0.14) v 0.09 (0.07-0.10) m/sec, P =.0017; e'/a': 1.20 (1.00-1.80) v 0.88 (0.71-1.05), P <.0001; E/e': 6.54 (4.75 7.14) v 8.89 (7.55-10.75), P <.0001, respectively). Within the hypertensive cohort, only TIMP-1 levels correlated with LV mass (r = 0.271, P =.024), LV mass index (r = 0.323, P =.007), and tissue Doppler parameters of diastolic dysfunction, including e' (r = -0.338, P =.005), a' (r = -0.350, P =.005), and E/e' (r = 0.334, P =.005). CONCLUSIONS: TIMP-1 is thought to increase tissue concentrations of collagen type I by preventing its breakdown by MMPs. Our findings therefore add weight to a hypothesis suggesting that TIMP-1 may be a key mediator of LV diastolic dysfunction through definition of ventricular matrix composition. PMID- 15363819 TI - Downregulation of vascular matrix metalloproteinase inducer and activator proteins in hypertensive patients. AB - BACKGROUND: Peripheral vasculature undergoes extensive vascular remodeling in the hypertensive state. Regulation of extracellular matrix turnover by the matrix metalloproteinase (MMP) system is an important step in the vascular remodeling process. However, the expression pattern of the vascular MMP system in human hypertension remained unknown. METHODS AND RESULTS: Internal mammary artery specimens were obtained from normotensive (n = 13) and hypertensive (n = 19) patients undergoing coronary artery bypass grafting surgery. Zymographic analysis indicated a threefold decrease in total gelatinolytic activity of MMP-2 and MMP-9 in hypertension. MMP-1 activity was also decreased by fourfold without a significant change in protein levels. Tissue levels of extracellular matrix inducer protein (EMMPRIN), MMP activator protein (MT1-MMP), MMP-1, MMP-2, and MMP 9, as well as tissue inhibitors of MMPs (TIMP-1 and TIMP-2) were assessed by immunoblotting and yielded a significant decrease in MMP-9, EMMPRIN, and MT1-MMP levels in hypertension. In addition, measurement of plasma markers of collagen synthesis (procollagen type I amino-terminal propeptide [PINP]) and collagen degradation (carboxy-terminal telopeptide of collagen type I [ICTP]) indicated no difference in PINP levels but suppressed degradation of collagen in hypertension. Evaluation of profibrotic growth factors demonstrated higher levels of fibroblast growth factor (FGF)-2 in tissue preparations from hypertensive patients but no difference in transforming growth factor-beta1 levels. CONCLUSIONS: These findings demonstrate that not only MMP-1 and MMP-9, but MMP inducer and activator proteins are also downregulated in the hypertensive state. Augmented FGF-2 levels may contribute to parallel decreases in MMP activity and MMP induction system resulting in enhanced collagen deposition in hypertension. PMID- 15363820 TI - Erythropoietin and obstructive sleep apnea. AB - OBJECTIVE: We tested the hypothesis that repetitive severe hypoxemia resulting from obstructive sleep apnea would increase serum erythropoietin, and that this increase would be attenuated by effective treatment of obstructive sleep apnea. METHODS: We studied healthy untreated patients with obstructive sleep apnea (18 severe and 10 very mild) before and after acute treatment with continuous positive airway pressure, and 12 healthy control subjects free of obstructive sleep apnea. RESULTS: Baseline erythropoietin levels before sleep were similar in the obstructive sleep apnea and control groups. However, erythropoietin levels increased (by 20%, P =.037) in patients with severe obstructive sleep apnea after 3.5 hours untreated (lowest O2, 77% +/- 3%), and decreased after 4 hours of continuous positive airway pressure treatment (P =.001). Erythropoietin responses in patients with severe obstructive sleep apnea were different (F = 4.0, P =.03) from controls, in whom erythropoietin levels remained stable throughout the night (P =.94). Erythropoietin responses were similar in very mild obstructive sleep apnea and controls (P =.58). CONCLUSIONS: Our results indicate that untreated severe obstructive sleep apnea results in increased erythropoietin, which decreases after continuous positive airway pressure treatment. Increased erythropoietin may be a potential reversible mechanism to explain the association between obstructive sleep apnea and cardiovascular disease. PMID- 15363821 TI - Relationship between daytime sleepiness and blood pressure in healthy older adults. AB - BACKGROUND: Some sleep disorders have been linked to hypertension, but few studies have examined the relationship between daytime sleepiness and blood pressure (BP). This study attempted to determine whether scores on a short questionnaire assessing daytime sleepiness (Epworth Sleepiness Scale [ESS]) were associated with BP and could be used to predict hypertension after 5 years in healthy older adults who had not previously been diagnosed with hypertension. METHODS: A group of 157 healthy men and women 55 to 80 years of age completed an extensive medical examination, a series of psychosocial tests, and two 24-h ambulatory BP sessions. After 5 years the procedures were repeated in 133 (85%) of the subjects. Psychosocial variables and BP were compared in subjects scoring high (score of > or = 10) and low (< 10) on the ESS. RESULTS: Compared to individuals with low ESS sores, those scoring high had increased casual and sleep BP as well as higher systolic BP levels and diastolic BP variability during waking hours, and reported higher levels of anger, depression, anxiety, and intensity of psychological symptoms as well as lower defensiveness. Individuals with high ESS scores were more likely to be diagnosed with hypertension 5 years later. Groups with high and low ESS scores did not differ significantly on any other variables. CONCLUSIONS: The ESS, a simple measure of daytime sleepiness, identified individuals at risk for hypertension. Future studies should investigate the possibility that diagnosis and treatment of daytime sleepiness could aid in BP reduction and ultimately in decreased morbidity and mortality from cardiovascular disorders. PMID- 15363822 TI - Rationale and design of the avoiding cardiovascular events through combination therapy in patients living with systolic hypertension (ACCOMPLISH) trial: the first randomized controlled trial to compare the clinical outcome effects of first-line combination therapies in hypertension. AB - Reducing blood pressure (BP) to target levels is a major priority in preventing clinical events in hypertension. Typically this requires more than one drug, and recent guidelines on hypertension management therefore recommend starting with combination treatment in many patients. Diuretics have often been part of such therapy, usually paired with angiotensin converting enzyme (ACE) inhibitors or similar agents; but calcium channel blockers are also highly efficacious in reducing BP when combined with ACE inhibitors. In addition, these drug classes, separately and in combination, appear to have vasculoprotective properties. Because the primary goal of treating hypertension is to enhance survival and reduce cardiovascular outcomes, the Rationale and Design of Avoiding Cardiovascular events through COMbination therapy in Patients LIving with Systolic Hypertension (ACCOMPLISH) trial is designed as the first blinded and randomized study to prospectively compare the effects on these endpoints of two antihypertensive combinations, benazapril/hydrochlorothiazide (force titrated to 40/12.5 mg) and amlodipine besylate/benazapril (force titrated to 5/40 mg). The doses can be further titrated to 40/25 mg or 10/40 mg, and other classes of drugs can be added as needed for optimal BP control. The primary study endpoint is a composite of cardiovascular mortality and morbidity. The study will be performed in hypertensive patients (systolic BP > or = 160 mm Hg or currently on antihypertensive therapy) with risk factors for cardiovascular events (prior events, target organ damage, kidney disease, or diabetes). A total of 6300 subjects will be randomized to each group with the expectation that a total of 1642 primary endpoints will occur during a 5-year period, providing 90% power to detect the 15% relative reduction in events (alpha = 0.05) hypothesized to favor the amlodipine besylate/benazapril group. The ACCOMPLISH study will be performed in the United States and Europe. The first patient was randomizedduring 2003, and the trial should conclude in 2008. PMID- 15363823 TI - Plasma asymmetric dimethylarginine and coronary and peripheral endothelial dysfunction in hypertensive patients. AB - BACKGROUND: The attenuation of coronary flow reserve (CFR) and endothelium mediated vasodilation of the brachial artery (EMV-BA) have been frequently reported in hypertensive patients. The present study investigated the link between CFR and EMV-BA in hypertensive patients. We hypothesized that changes in serum asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, and concomitant insulin resistance may be underlying factors connecting the two pathologic alterations. METHODS: A total of 75 patients (30 men and 45 women, 61.5 +/- 10.1 years of age) with essential hypertension and without coronary artery disease and diabetes mellitus were included in the study. Measurements of CFR were made using adenosine-triphosphate stress transthoracic Doppler echocardiography, and forearm EMV-BA was measured by venous occlusion strain gauge plethysmography. A plasma ADMA assay and a 75-g oral glucose tolerance test were also performed. RESULTS: Average CFR and EMV-BA values were 2.54 +/- 0.63 and 86.0 +/- 54.7%, respectively. A significant correlation was found between CFR and EMV-BA (r = 0.493, P <.001). Both CFR and EMV-BA were also significantly correlated with age and plasma ADMA concentration, but were not correlated with insulin resistance, plasma insulin, or left ventricular mass. Multiple regression analysis revealed that ADMA was the only statistically independent parameter associated with CFR and EMV-BA. CONCLUSIONS: The similar deterioration in endothelial function in coronary and peripheral vascular territories may be mainly due to increased plasma ADMA concentration. Plasma ADMA appears to play a major role in endothelial dysfunction in hypertensive patients, independent of insulin resistance or left ventricular hypertrophy. PMID- 15363824 TI - Factors related to the impact of antihypertensive treatment in antioxidant activities and oxidative stress by-products in human hypertension. AB - The objective was to study factors related to the changes induced by antihypertensive treatment on oxidative status, antioxidant activities, and reactive oxygen species by-products in whole blood and mononuclear peripheral cells. Eighty-nine hypertensive patients (mean age 46 years, 46 men, average 24-h blood pressure 139/88 mm Hg, body mass index 29) were included. After 3 months of nonrandomized allocation to antihypertensive treatment (20 nonpharmacologic, 36 beta-blockers, 33 angiotensin receptor blocker), oxidized/reduced glutathione ratio and malondialdehyde were significantly reduced, and the activity of superoxide dismutase, catalase, and glutathione peroxidase was significantly increased in both whole blood and peripheral mononuclear cells. The content of damaged base 8-oxo-2'-deoxyguanosine in nuclear and mitochondrial DNA in hypertensive subjects was also significantly reduced during the antihypertensive treatment. In a group of 42 subjects, the oxidative stress was further reduced and the antioxidant enzyme activities further increased after 12 months of antihypertensive treatment. The changes were independent of the kind of antihypertensive treatment. In conclusion, antihypertensive treatment improved the increased oxidative stress and the decreased antioxidant mechanisms. It is independent of the type of treatment and the beneficial effect of treatment increases over time. PMID- 15363825 TI - Calcium channel blockade to prevent stroke in hypertension: a meta-analysis of 13 studies with 103,793 subjects. AB - BACKGROUND: The possibility that specific antihypertensive treatments may prevent the occurrence of stroke more effectively than other treatments remains unproved. We undertook a meta-analysis to assess whether calcium channel blockers (CCBs) are associated with a lesser risk of stroke as compared with other antihypertensive drugs. METHODS: Through Medline we identified 13 major studies conducted in hypertensive subjects for a total of 103,793 subjects. Overall, there were 4040 incident cases of stroke, 1789 among 43,053 subjects randomized to CCBs and 2251 among 60,740 subjects randomized to different antihypertensive drugs. RESULTS: Considering all 13 trials, a pooled reduction in the risk of stroke was observed among subjects allocated to CCBs (odds ratio 0.90, 95% confidence interval [95% CI] 0.84-0.96; P =.002). The risk of stroke was significantly lower among subjects allocated to dihydropyridine CCBs than among those randomized to alternative drugs (odds ratio 0.90, 95% CI 0.84-0.97; P =.006), whereas the effect of non-dihydropyridine CCBs did not achieve significance (odds ratio 0.92, 95% CI 0.81-1.04). In a meta-regression analysis of these trials, the protection from stroke conferred by CCBs appeared unrelated to the degree of systolic blood pressure reduction. CONCLUSIONS: These findings suggest that CCBs decrease the risk of stroke more effectively than other treatments in patients with essential hypertension and that such an effect might not be completely explained by a better antihypertensive response. Calcium channel blockers should be considered in hypertensive subjects at increased risk of stroke. PMID- 15363826 TI - Effect of amlodipine-atorvastatin combination on fibrinolysis in hypertensive hypercholesterolemic patients with insulin resistance. AB - BACKGROUND: The aim of this study was to evaluate the effect of the amlodipine atorvastatin combination on plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) activity in hypercholesterolemic, hypertensive patients with insulin resistance. METHODS: The study population included 45 patients, aged 41 to 70 years, with mild to moderate essential hypertension (diastolic blood pressure [BP] > or = 95 and < or = 105 mm Hg), hypercholesterolemia (total cholesterol > 200 and < 350 mg/dL), and insulin resistance (HOMA index > 2.5) After a 4-week wash-out period, they were randomized to amlodipine (5 mg) or atorvastatin (20 mg) or their combination at the same oral dosage for 12 weeks in three cross-over periods each separated by a 4-week placebo period (3 by 3 latin square design). At the end of the placebo wash-out and of each treatment period, office BP, total cholesterol, PAI-1, and t PA activity were evaluated. RESULTS: The amlodipine-atorvastatin combination, in addition to the expected hypocholesterolemic effect, produced: 1) a greater decrease in PAI-1 activity (-10.2 U/mL, P <.01 v placebo) and an even greater increase in t-PA activity (+0.26 U/mL, P <.01 v placebo) than amlodipine (-0.5 U/mL for PAI-1, P = not significant; +0.17 U/mL for t-PA, P <.01 v placebo) and atorvastatin alone (respectively, -9.9 U/mL, P <.01 v placebo and +0.08 U/mL, P <.05 v placebo); and 2) a greater systolic BP/diastolic BP mean reduction (-22/17 mm Hg, P <.005 v placebo) than amlodipine (-18/14 mm Hg, P <.01 v placebo) and atorvastatin alone (-2.8/3.8 mm Hg, P <.05 v placebo only for diastolic BP). CONCLUSIONS: The positive effect on fibrinolytic balance and BP control observed suggests that in hypertensive, hypercholesterolemic patients with impaired fibrinolysis, the combination of amlodipine and atorvastatin could be the treatment of choice. PMID- 15363827 TI - Association of inappropriate left ventricular mass with systolic and diastolic dysfunction: the HyperGEN study. AB - BACKGROUND: Left ventricular mass (LVM) exceeding values appropriate for individual workload is associated with concentric LV geometry and systolic dysfunction at both the midwall and chamber levels, even independently of clearcut LV hypertrophy, and is a marker of adverse prognosis. No information is available on whether inappropriate LVM is associated with alterations of LV diastolic properties in unselected population-based studies. METHODS: We examined 1513 hypertensive participants from the HyperGEN population who were without prevalent cardiovascular disease. The LVM prediction from stroke work (systolic blood pressure x Doppler stroke volume), sex, and height (in meters2.7) was derived in 210 nonobese, normal individuals. Observed/predicted LVM >133% defined inappropriate LVM. RESULTS: Participants with inappropriate LVM (229 subjects [15%]) exhibited higher LV dimensions and relative wall thickness, lower ejection fraction, midwall shortening and cardiac output, and prolonged isovolumic relaxation time (IVRT; P < or = .001). Other diastolic abnormalities were not associated with inappropriate LVM. After accounting for demographic and hemodynamic confounders, IVRT was directly related to excess LVM, whereas deceleration time of E velocity was negatively related to excess LVM. Nonechocardiographic correlates of prolonged relaxation included aging, African American ethnicity, male gender, and diabetes. CONCLUSIONS: The LVM exceeding the compensatory needs for workload is associated with delayed LV relaxation as well as mild midwall and chamber systolic dysfunction, independently of demographic, clinical, and hemodynamic confounders. Inappropriately high LVM also correlates with features of increased myocardial stiffness, coexisting with evidence of prolonged LV relaxation. PMID- 15363828 TI - Genome scan for hypertension in nonobese African Americans: the National Heart, Lung, and Blood Institute Family Blood Pressure Program. AB - BACKGROUND: Obesity is an important risk factor for hypertension, but epidemiologic studies provide evidence for the development of hypertension independent of obesity. In addition, the search for hypertension susceptibility genes should prove more informative when applied to a homogeneous subset of patients, such as those that are not obese. For this reason, we sought to identify genomic regions influencing susceptibility to hypertension in a nonobese sample of hypertensive African American families. METHODS: A genome-wide linkage scan was performed in a sample of 275 African American hypertensive families containing two or more nonobese (body mass index, < or = 30 kg/m2) individuals recruited by Networks of the Family Blood Pressure Program (FBPP). RESULTS: The best evidence for linkage of hypertension among the FBPP African American families was found on chromosome 2 (log of the odds [LOD]= 3.59 at 230 cM). All other chromosomes contained LOD scores less than 2. The African American sibships from the GENOA Network appear to largely contribute to the evidence for linkage on chromosome 2 (LOD = 4.07 at 233 cM). CONCLUSIONS: Significant evidence for linkage to hypertension in nonobese African American families was identified on chromosome 2q. These results suggest the presence of genes influencing susceptibility to adiposity-independent hypertension. PMID- 15363829 TI - Genome-wide linkage analyses for age at diagnosis of hypertension and early-onset hypertension in the HyperGEN study. AB - A genome-wide scan was performed to identify chromosomal regions related to age at diagnosis of hypertension and to early-onset hypertension in white and African American families from the Hypertension Genetic Epidemiology Network (HyperGEN). Age at diagnosis of hypertension was reported by participants at recruitment and standardized residuals adjusted for sex, study center, and body mass index were created. Participants were classified as having early-onset hypertension if their reported age at diagnosis was before 45 years in whites or before 35 years in African Americans. Variance component linkage analysis was performed for age at diagnosis and an affected sibpair linkage analysis was performed for early-onset hypertension, both implemented in GENEHUNTER. In whites, the heritability of diagnosis age was estimated to be 35% and the maximum LOD score was found on chromosome 1 at 123 cM (LOD = 1.48). The maximum LOD score for early-onset hypertension was located on chromosome 18 at 69 cM (LOD = 1.21). In African Americans, the heritability of age of diagnosis was estimated to be 42% and the maximum LOD scores were found on chromosome 4 at 120 cM (LOD = 2.44) and on chromosome 15 at 60 cM (LOD = 2.31). The maximum LOD for early-onset hypertension in African Americans was also on chromosome 4 at 153 cM (LOD = 2.05) and overlies the mineralocorticoid receptor. Although these results report modest LOD scores, several of these loci have been previously reported to be linked to hypertension and blood pressure, lending further support that genes related to the risk of hypertension may be at these loci. PMID- 15363830 TI - Low-density lipoprotein particle size and coronary atherosclerosis in subjects belonging to hypertensive sibships. AB - BACKGROUND: Dyslipidemia in hypertensive sibships may be characterized by atherogenic small low-density lipoprotein (LDL) particles. Whether LDL particle size is associated with the extent of coronary atherosclerosis in hypertensive sibships is unknown. METHODS: Subjects (n = 792, mean age 62 years, 60% women) were ascertained through sibships containing at least two individuals with essential hypertension diagnosed before age 60 years. The LDL particle size was measured by polyacrylamide gel electrophoresis. Coronary artery calcium (CAC) was measured noninvasively by electron beam computed tomography, and CAC score was calculated using the method of Agatston et al. Sex-specific multiple regression models were used to assess independent predictors of LDL particle size and the association of LDL particle size with CAC. RESULTS: In all, 76% of women and 77% of men were hypertensive. In each sex, independent predictors of smaller LDL particle size were total cholesterol, triglycerides, and lower HDL cholesterol. In women, greater age was an additional predictor of smaller LDL particle size. After adjustment for age and statin use, LDL particle size was significantly associated with the amount of CAC in women but not in men. After further adjustment for HDL cholesterol, triglycerides, diabetes, smoking, and hypertension, LDL particle size was not independently associated with CAC in either sex. CONCLUSIONS: After adjustment for age and statin use, LDL particle size was found to be significantly related to CAC quantity in women but not in men belonging to hypertensive sibships. In women, LDL particle size may mediate some of the atherogenic effects of low-HDL cholesterol-high-triglyceride dyslipidemia, but does not appear to be independently associated with the extent of coronary atherosclerosis in either sex. PMID- 15363831 TI - Reactive oxygen species in hypertension; An update. AB - Hypertension is associated with an elevation of reactive oxygen species (ROS) and frequently also with an impairment of endogenous antioxidant mechanisms. Experimental manipulation of the redox state in vivo shows that ROS can be a cause of hypertension. During the development of the disease, ROS are generated by endogenous sources, notably the NADPH oxidase enzyme family and uncoupled nitric oxide synthase, due to a mutual reinforcement between ROS and humoral factors. The ROS affect multiple tissues, either directly or through nitric oxide depletion. In the vasculature, they induce contraction and endothelial dysfunction. In blood vessels and myocardium, they cause hypertrophic remodeling. In the kidneys, ROS promote salt reabsorption, decrease glomerular filtration, and lead to tissue damage. Finally, they also increase efferent sympathetic activity from the central nervous system. Progress in our understanding of the mechanisms of ROS formation and their plethora of pathophysiologic effects is expected to lead from simple antioxidant therapy to specific antihypertensive treatments. PMID- 15363832 TI - Relationship between white blood cell count and incident hypertension. PMID- 15363834 TI - Three-dimensional first trimester fetal volumetry: comparison with crown rump length. AB - BACKGROUND: Ultrasonographic volumetry measurements of human fetus have become possible using three-dimensional ultrasound systems. OBJECTIVE: To evaluate the weekly increase of fetal volume during the first trimester of normal pregnancies compared to the crown rump length and creating a first trimester fetal volume nomogram. METHODS: Crown rump length and three-dimensional ultrasonographic volumetry measurements performed on 72 first trimester fetuses using virtual organ computer aided analysis (VOCAL). RESULTS: A significant direct correlation (r = 0.939) was found between the calculated fetal volumes and crown rump lengths. A 6-12 weeks gestation fetal volume nomogram was proposed. CONCLUSION: Fetal volume database in the first trimester may serve as a reference table for diagnosis of early pregnancy failure. PMID- 15363835 TI - Growth patterns of human ovarian volume during intrauterine and postnatal organogenesis. AB - BACKGROUND: Ovarian volumes during pre-pubertal, reproductive or menopausal life in a healthy female have been studied by in vivo and anatomical methods. In contrast, conclusive studies on normal human ovarian volume during intrauterine and postnatal organogenesis, when the folliculogenesis occurs, do not exist. AIM: To assess the growth patterns of ovarian volume during human ovarian definitive histogenesis using morphometric methods. SUBJECTS: Twenty-five left ovaries removed from fetuses, newborns and children with karyotype 46,XX, ranging from 15 to 74 weeks of development. OUTCOME MEASURES: The ovaries were completely cut obtaining serial sections. Ovarian volume was calculated both with the ellipsoid method, using its length, maximum and minimum diameters, and the Cavalieri quantitative morphometric method. The age-related pattern of ovarian volume was studied using linear regression analyses. RESULTS: Ovarian volume calculated with the Cavalieri method increased from 16 mm3 at the 15th week of development to over 300 mm3 at the end of the process, with a 20-fold increase compared to the initial volume. Ovarian volume calculated with the ellipsoid method ranged between 25 to over 380 mm3, with a 15-fold increase. On average, ellipsoidal volume over-estimated the true Cavalieri volume (mean difference -14%, S.D. 29, p=0.014, Student's t-test for paired samples). Linear regression curves were developed to predict ovarian volume at any age. Overall, the approximation was better for Cavalieri volume (approximately, 72% of variance explained by the linear regression) than for ellipsoid volume (59% of explained variance). CONCLUSIONS: During the intrauterine and postnatal organogenesis, the normal growth rate of human ovary was almost linear. The calculation of ovarian volume using Cavalieri method does not depend on assumptions about the organ shape and it appears to be more appropriate. PMID- 15363836 TI - Movement and motor development in ELBW infants at 1 year is related to cognitive and motor abilities at 4 years. AB - A relationship between motor ability and cognitive performance has been previously reported. This study aimed to investigate the association between movement and cognitive performance at 1 and 4 years corrected age of children born less than 1000 g, and whether developmental testing of movement at 1 year is predictive of cognitive performance at 4 years. Motor development was assessed at both ages using the neurosensory motor developmental assessment (NSMDA) and motor development was classified as normal, or minimal, mild, moderate-severe dysfunction. Cognitive performance was assessed on the Griffith Mental Developmental Scale at 1 year and McCarthy Scales of Children's Abilities at 4 years. Subjects included 198 children of birthweight less than 1000 g. Of these 132 children returned for follow-up at the corrected ages of both 1 and 4 years. The 66 children not included had a slight increase in gestational age, while the mothers were younger and had a lower level of education. A significant association was found between NSMDA group classification at 1 year and cognitive performance at both 1 and 4 years (p<0.001; p<0.0001) and between the subscales of each test (1 year, p<0.0001; 4 years, p<0.001). Group classification of motor development at 1 year was predictive of cognitive performance at 4 years (p<0.0001) and this was independent of biological and social factors and presence of cerebral palsy (CP). The findings support a close link between motor and cognitive development in children born <1000 g and emphasise the advantage of detailed assessment of movement at 1 year. PMID- 15363837 TI - Influence of maternal tobacco smoking during pregnancy on uterine, umbilical and fetal cerebral artery blood flows. AB - BACKGROUND: Cigarette smoking during pregnancy is a major cause of adverse perinatal outcome but effects on critical fetal and maternal circulations remain unclear. AIMS: To determine (1) the influence of habitual maternal cigarette smoking on blood flow velocities in uterine, umbilical and fetal middle cerebral arteries, and (2) the time-course of changes in these flows after smoking a cigarette. SUBJECTS AND METHODS: In 74 women who smoked and 69 non-smoking controls we measured Doppler blood flow velocity waveforms in the uterine, umbilical and fetal middle cerebral (MCA) arteries; smokers were divided into light (<10 cigarettes/day, n=28) and heavy smokers (>10 cigarettes/day, n=45). RESULTS: In the uterine artery there was no significant difference between the systolic/diastolic (S/D) ratio in smoking and non-smoking women; however, in smokers, a diastolic notch was more frequently observed in the uterine artery waveform than in controls (p<0.05), suggestive of a greater resistance in the uterine vasculature. In the umbilical artery, the S/D ratio was significantly greater in smokers than in non-smokers. In the fetal MCA, the S/D ratio was higher in heavy smokers than in light smokers (p<0.05) indicative of greater cerebrovascular resistance. There were no significant correlations between the interval between smoking the last cigarette and making the Doppler measurements and the S/D ratios in any of the vessels. CONCLUSION: Maternal cigarette smoking is associated with evidence of chronically increased resistances in the uterine, umbilical and fetal middle cerebral arteries. PMID- 15363838 TI - Sleeping and waking state development in preterm infants. AB - BACKGROUND: Most studies of sleep-wake states of preterm infants have been cross sectional. Thus, the extent to which sleep-wake development occurs within individuals and how environmental factors affect the development of sleeping and waking is unclear. AIMS: This study examined the development of sleeping and waking during the preterm and early post-term periods and the effects of infant health and environmental characteristics. DESIGN: Longitudinal, descriptive design. PARTICIPANTS: 134 preterm infants at high risk for developmental problems because of birthweights under 1500 g or mechanical ventilation. OUTCOME MEASURES: Weekly 2-h behavioral observations were conducted from the time infants were no longer critically ill until 43 weeks post-conceptional age or discharge. A single follow-up observation was conducted 1-3 months later. RESULTS: Active sleep, large body movements and the percent of no REM during active sleep decreased with age, and quiet waking, active waking, quiet sleep and regularity of respiration in active sleep and quiet sleep increased. The state of sleep-wake transition increased until 40 weeks and then decreased after 43 weeks CA. Negative facial expressions showed a quadratic decrease over age. Active waking, active sleep, negative facial expressions and quiet sleep regularity showed a change of development after term. Infant characteristics, illness severity and medical treatments, the handling due to performing an EEG and hospital had only minor effects. CONCLUSIONS: Significant development of sleeping and waking occurs over the preterm period. Additional research is needed to determine how the change from the hospital to the home environment affects on these developmental trajectories. PMID- 15363839 TI - Follow up of children after antenatal treatment for alloimmune thrombocytopenia. AB - OBJECTIVE: To evaluate the long-term follow-up of surviving offspring after antenatal treatment for fetal or neonatal alloimmune thrombocytopenia (FNAIT). PATIENTS: Fifty children at risk of FNAIT were antenatally treated with maternal intravenous immunoglobulins (IVIG) (n=11), IVIG with intrauterine platelet transfusions (IUPT) (n=26) or IUPT alone (n=9). In four cases (n=4), only fetal blood sampling (FBS) was performed. One child died in the neonatal period and one was lost to follow up. METHODS: The remaining 48 children, aged 1.3-11.6 years (median 5.1 years), were given both general and neurological examinations and assessed on their development and susceptibility for infections or atopic constitution. In addition, immunoglobulin levels were measured in 17 infants, aged 5 years and older. RESULTS: Intracranial hemorrhage (ICH) was not observed. The general health and neurodevelopmental outcome in the children was comparable to a normal Dutch population. Children not exposed to maternal IVIG treatment had significantly more infections and hearing problems than children exposed to IVIG treatment or the normal population. Immunoglobulin G, A and M levels were within the normal range, independent of treatment and severity of FNAIT. A high IgE level was more frequently seen in children exposed to IVIG, but did not result in clinical consequences such as allergy or atopy. CONCLUSIONS: Antenatal treatment of children for FNAIT did not affect general health or neurodevelopmental outcome. In particular, exposure to IVIG in utero showed no adverse effect on the clinical outcome of these children. PMID- 15363840 TI - Analysis of a conserved duplicated tRNA gene in the mitochondrial genome of blowflies. AB - The mtDNA control region (CR) and flanking genes of the blowflies Chrysomya albiceps, Chrysomya megacephala and Chrysomya chloropyga (Calliphoridae) were characterized. The most unusual feature found was the presence of duplicated tRNA sequences corresponding to trnI and a portion of trnQ. The partially duplicated trnQ was very likely a pseudogene since most of the sequence of the typical insect trnQ gene was missing. In contrast, the trnI gene had a conserved primary sequence following the duplication event and may represent a functional copy. These results demonstrate the plasticity of the mtDNA molecule in Chrysomya, especially for tRNA genes and the adjacent control region sequences. PMID- 15363841 TI - The mitochondrial genome of the blowfly Chrysomya chloropyga (Diptera: Calliphoridae). AB - In view of the medical, sanitary and forensic importance of Chrysomya species, a knowledge of their nucleotide sequences would be useful for the molecular characterization of this genus, and would help in designing primers and in improving the molecular identification of Calliphoridae species. In this work, the mitochondrial genome of the blowfly Chrysomya chloropyga (Diptera: Calliphoridae) was completely sequenced. The entire mitochondrial DNA (mtDNA) molecule was 15,837 bp long and was sequenced using the shotgun approach. The overall nucleotide composition was heavily biased towards As and Ts, which accounted for 76.7% of the whole genome. The cox1 gene had a serine as the start codon, while incomplete termination codons mediated by tRNA signals were found for cox2, nd4 and nd5. The C. chloropyga genes were in the same order and orientation as the mitochondrial genome of other dipteran species, except for the occurrence of a 123 bp region that included a complete duplication of tRNA(Ile) and a partial duplication of tRNA(Gln) genes. C. chloropyga is the first species of Diptera with 23 tRNA genes instead of the usual 22 already described. A phylogenetic analysis showed a split of Brachycera into Calyptratae and Acalyptratae subdivisions. The complete sequence of C. chloropyga mtDNA described here will be a useful source of sequence information for general molecular and evolutionary studies in Diptera. PMID- 15363842 TI - Alternative polyadenylation and splicing of mRNAs transcribed from the human Sin1 gene. AB - Limited but significant sequence similarity has been observed between an uncharacterized human protein, SIN1, and the S. pombe SIN1, Dictyostelium RIP3 and S. cerevisiae AVO1 proteins. The human Sin1 gene has been automatically predicted (MAPKAP1; GenBank accession number ); however, this sequence appears to be incomplete. In this study, we have cloned and characterized the full-length human Sin1 mRNA and identified a highly conserved domain that defines the family of SIN1 orthologues, members of which are widely distributed in the fungal and metazoan kingdoms. We demonstrate that Sin1 transcripts can use alternative polyadenylation signals and describe a number of Sin1 splice variants that potentially encode functionally different isoforms. PMID- 15363843 TI - ASF/SF2-like maize pre-mRNA splicing factors affect splice site utilization and their transcripts are alternatively spliced. AB - Three ASF/SF2-like alternative splicing genes from maize were identified, cloned, and analyzed. Each of these genes (zmSRp30, zmSRp31, and zmSRp32) contains two RNA binding domains, a signature sequence SWQDLKD, and a characteristic serine/ariginine-rich domain. There is a strong structural similarity to the human ASF/SF2 splicing factor and to the Arabidopsis atSRp34/p30 proteins. Similar to ASF/SF2-like genes in other organisms, the maize pre-mRNA messages are alternatively spliced. They are differentially expressed in maize tissues with relatively uniform levels of zmSRp30 and zmSRp31 messages being observed throughout the plant, while zmSRp32 messages preferentially accumulated in the meristematic regions. Overexpression of zmSRp32 in maize cells leads to the enhanced selection of weak 5' intron splice sites during the processing of pre mRNA molecules. Overexpression of the zmSRp31 or zmSRp32 gene affects regulation of wheat dwarf virus rep gene pre-mRNA splicing, presumably by interacting with the weak 5' splice site, CCGU. Our results suggest that the described genes are functional homologues of the human ASF/SF2 alternative splicing factor and they indicate a diversity of the ASF/SF2-like alternative splicing factors in monocot plant cells. PMID- 15363844 TI - Comparative analysis of ATRX, a chromatin remodeling protein. AB - The ATRX protein, associated with X-linked alpha-thalassaemia, mental retardation and developmental abnormalities including genital dysgenesis, has been proposed to function as a global transcriptional regulator within a multi-protein complex. However, an understanding of the composition and mechanics of this machinery has remained elusive. We applied inter-specific comparative analysis to identify conserved elements which may be involved in regulating the conformation of chromatin. As part of this study, we cloned and sequenced the entire translatable coding region (7.4 kb) of the ATRX gene from a model marsupial (tammar wallaby, Macropus eugenii). We identify an ATRX ancestral core, conserved between plants, fish and mammals, comprising the cysteine-rich and SWI2/SNF2 helicase-like regions and protein interaction domains. Our data are consistent with the model of the cysteine-rich region as a DNA-binding zinc finger adjacent to a protein binding (plant homeodomain-like) domain. Alignment of vertebrate ATRX sequences highlights other conserved elements, including a negatively charged mammalian sequence which we propose to be involved in binding of positively charged histone tails. PMID- 15363845 TI - Molecular evolution of epididymal lipocalin genes localized on mouse chromosome 2. AB - We previously identified two murine secretory proteins, mE-RABP(Lcn5) and mEP17(Lcn8), belonging to the lipocalin family and specifically expressed in the epididymis. The genes are contiguous and localized on mouse chromosome 2. We now show that five other related lipocalin genes, Lcn9, Lcn10, Lcn11, Lcn12, and Lcn13, that evolved by in situ tandem duplication are present on the same locus. Lcn9, Lcn10, Lcn12, and Lcn13 genes, like Lcn5 and Lcn8 genes, are specifically expressed in the mouse epididymis. However, each gene has a distinct spatial expression within the epididymis and different regulation. Analysis of the human genome sequence shows the presence of genes encoding lipocalins with genomic organization, chromosomal arrangement, and orientation similar to that of the corresponding murine genes, indicating that the epididymal cluster is evolutionary conserved. A phylogenetic analysis of the new epididymal proteins reveals their spread position in the lipocalin protein family tree. This suggests the preservation of the regulatory sequences, while protein sequences have greatly diverged, reflecting functional diversity and possibly multifunctionality. In terms of the cluster ancestry, epididymal expression possibly appeared in a PGDS-like lipocalin in amniotes, and the duplications generating the cluster occurred at least in the common ancestor of rodents and primates. The presence and conservation of a cluster of five genes encoding epididymal lipocalins, differently regulated and regionalized in the epididymis, strongly suggests that these proteins may play an important role for male fertility. PMID- 15363846 TI - The chicken telomerase reverse transcriptase (chTERT): molecular and cytogenetic characterization with a comparative analysis. AB - Telomerase activity is essential for maintaining the termini of linear chromosomes. Telomerase consists of both a RNA and a specialized reverse transcriptase. Our objective for this study was to determine the molecular and cytogenetic features of the chicken telomerase reverse transcriptase (chTERT) gene and protein. The TERT mRNA from gastrula stage embryos was found to be 4497 bp in length, translating into a protein of 1346 amino acids (aa). The chTERT protein shares 45% aa identity with human TERT (hTERT). A distinctive feature of chTERT, as compared to human and other vertebrate TERTs, is the larger size of the protein due mainly to a considerably longer N-terminal flexible linker region (144 aa longer than in human). Chicken TERT was mapped to chromosome 2q21 near an interstitial telomere site. Several transcription factor binding motifs in the 5' flanking/promoter region of chTERT were similar to those found associated with hTERT (E-box, Ik1, MAZ, Sp1 sites), whereas several c-Myb sites were found associated with chTERT only and c-Ets-2 and WT1 were associated with hTERT only. Results presented here should promote structure-function studies of chTERT, as well as contribute to the comparative analysis of TERT regulation and function in vertebrates utilizing the telomere clock mechanism to different degrees. PMID- 15363847 TI - Polymorphism in the bovine BOLA-DRB3 upstream regulatory regions detected through PCR-SSCP and DNA sequencing. AB - In the present work, we describe through polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing the polymorphism within the URR-BoLA-DRB3 in 15 cattle breeds. In total, seven PCR-SSCP defined alleles were detected. The alignment of studied sequences showed six polymorphic sites (four transitions, one transversion and one deletion) in the interconsensus regions of the BoLA-DRB3 upstream regulatory region (URR), while the consensus boxes were invariant. Five out of six detected polymorphic sites were of one nucleotide substitution in the interconsensus regions. It is expected that these mutations do not affect significantly the level of expression. In contrast, the deletion observed in the sequence between CCAAT and TATA boxes could have some effect on affinity interactions between the promoter region and the transcription factors. The URR-BoLA-DRB3 DNA analyzed sequences showed moderate level of nucleotide diversity, high level of identity among them and were grouped in the same clade in the phylogenetic tree. In addition, the phylogenetic tree, the similarity analysis and the sequence structure confirmed that the fragment analyzed in this study corresponds to the URR-BoLA-DRB3. The functional role of the observed polymorphic sites among the regulatory motifs in bovine needs to be analyzed and confirmed by means of gene expression assays. PMID- 15363848 TI - Secreted subtilisin gene family in Trichophyton rubrum. AB - Secreted proteases constitute potential virulence factors of dermatophytes. A total of seven genes encoding putative serine proteases of the subtilisin family (SUB) were isolated in Trichophyton rubrum. Based on sequence data and intron exon structure, a phylogenetic analysis of subtilisins from T. rubrum and other fungi revealed a presumed ancestral lineage comprising T. rubrum SUB2 and Aspergillus SUBs. All other SUBs (SUB1, SUB3-7) are dermatophyte-specific and have apparently emerged more recently, through successive gene duplication events. We showed that two subtilisins, Sub3 and Sub4, were detected in culture supernatants of T. rubrum grown in a medium containing soy protein as a sole nitrogen source. Both recombinant enzymes produced in Pichia pastoris are highly active on keratin azure suggesting that these proteases play an important role in invasion of keratinised tissues by the fungus. The set of deduced amino acid sequences of T. rubrum SUB ORFs allowed the identification of orthologous Subs secreted by other dermatophyte species using proteolysis and mass spectrometry. PMID- 15363849 TI - High levels of RNA editing in a vascular plant chloroplast genome: analysis of transcripts from the fern Adiantum capillus-veneris. AB - We sequenced transcripts from all putative genes for proteins, rRNAs, and a selection of gene-encoding tRNAs in the chloroplast genome of the fern Adiantum capillus-veneris. We detected 350 RNA editing sites when the cDNA sequence was compared to that of the genomic DNA. Of these sites, 10% were U-to-C edits and 90% were C-to-U edits. RNA editing created 19 new start codons, three new stop codons, and "repaired" 26 internal stop codons. Of the 332 editing sites that altered a codon, 26% were in the first codon position, 68% in the second, and 6% in the third. We also detected 21 silent edits, as well as 19 edits that were in untranslated regions, including introns and the anticodon of tRNA(Leu). The latter edit provided a tRNA that is not otherwise encoded in this genome and accounts for a heavily used leucine codon. The level of RNA editing in this fern is more than ten times that of any other vascular plant examined across an entire chloroplast genome. A previous study found even higher levels of editing in a hornwort (942 sites). This suggests that the relatively low levels of editing in seed plants (less than 0.05%) may not be typical for land plants, and that RNA editing may play a major role in chloroplast genome processing. Additionally, we found that 53 editing sites in A. capillus-veneris are homologous to editing sites in the hornwort, and some other land plants. This implies that a major component of RNA editing sites have been conserved for hundreds of millions of years. PMID- 15363850 TI - Identification of allelic variants of the bovine immune regulatory molecule CEACAM1 implies a pathogen-driven evolution. AB - Carcinoembryonic antigen-related cell adhesion molecule (CEACAM1), the primordial member of the carcinoembryonic antigen (CEA) family, functions as a MHC independent natural killer (NK) cell inhibitory receptor, regulates T and B cell proliferation, and induces dendritic cell (DC) maturation. Despite these fundamental functions, CEACAM1 and most of the CEA family members differ significantly in primates and rodents. A number of diverse murine and human pathogens use CEACAM1 as a cellular receptor, indicating that the observed species-specific differences are the result of divergent molecular pathogen/host coevolution. To gain deeper insight into its evolution and function, we cloned CEACAM1 cDNA from cattle as a representative of a third mammalian order. Bovine CEACAM1 differs considerably from rodent and primate CEACAM1 due to deletion of the B domain exon which was most likely caused by insertion of LINE/SINE sequences and reveals alternative splicing within the transmembrane exon. However, the characteristic long and short isoforms exist which contain or lack the typical immunoreceptor tyrosine-based inhibitory motifs (ITIM) in their cytoplasmic tails, respectively. Bovine peripheral blood lymphocytes (PBL) express only ITIM-containing CEACAM1 isoforms, and upregulate their expression upon stimulation, suggesting an inhibitory function in these cells. As found in rodents, two clearly distinct CEACAM1 alleles exist in cattle. In the a allele, a unique deletion of three amino acids is found in the N domain, which is important for pathogen binding in mice and humans. This is consistent with the notion that CEACAM1 serves or has served as a pathogen receptor in cattle. PMID- 15363851 TI - Carboxylic acids permeases in yeast: two genes in Kluyveromyces lactis. AB - Two new genes KlJEN1 and KlJEN2 were identified in Kluyveromyces lactis. The deduced structure of their products is typical of membrane-bound carriers and displays high similarity to Jen1p, the monocarboxylate permease of Saccharomyces cerevisiae. Both KlJEN1 and KlJEN2 are under the control of glucose repression mediated by FOG1 and FOG2, corresponding to S. cerevisiae GAL83 and SNF1 respectively, and KlCAT8, proteins involved in glucose signalling cascade in K. lactis. KlJEN1, but not KlJEN2, is induced by lactate. KlJEN2 in contrast is expressed at high level in ethanol and succinate. The physiological characterization of null mutants showed that KlJEN1 is the functional homologue of ScJEN1, whereas KlJEN2 encodes a dicarboxylic acids transporter. In fact, KlJen1p [transporter classification (TC) number: 2.A.1.12.2.] is required for lactate uptake and therefore for growth on lactate. KlJen2p is required for succinate transport, as demonstrated by succinate uptake experiments and by inability of Kljen2 mutant to grow on succinate. This carrier appears to transport also malate and fumarate because the Kljen2 mutant cannot grow on these substrates and the succinate uptake is competed by these carboxylic acids. We conclude that KlJEN2 is the first yeast gene shown to encode a dicarboxylic acids permease. PMID- 15363852 TI - Natural antisense transcripts of HIF-1alpha are conserved in rodents. AB - A natural antisense transcript (aHIF), which sequence is strictly complementary to the 3' untranslated region (3'UTR) of HIF-1alpha mRNA, has been identified in human and shown to be overexpressed in renal carcinomas. We searched for aHIF in different rodent tissues. Two candidate expressed sequence tag (EST) were identified in silico and their PCR products (1.1 and 1.0 kb) were cloned and sequenced in mouse and rat, respectively. These transcripts were rigorously complementary to the 3'UTR of rodent HIF-1alpha mRNA and were broadly expressed in all mouse and rat tissues we tested. The conservation of aHIF in rodents underlined its potential importance in cell regulations. Therefore the responses of aHIF and HIF-1alpha transcripts were investigated in various types of hypoxic conditions. In freshly isolated rat renal tubules, aHIF RNA level was increased by acute hypoxia and low in normal supply of oxygen. In a rat strain raised in chronic hypobaric altitude hypoxia, aHIF transcript was greatly induced in the oxidative-type soleus and heart muscles of 3 month-old animals. By contrast, in the glycolytic-type extensor digitorum longus muscle aHIF transcript amount was lowered by hypoxia whereas HIF-1alpha transcript was highly expressed. In brain, where oxidative glycolysis takes place, HIF-1alpha mRNA and its antisense transcript levels were high and not significantly changed by altitude. Tumour cell lines cultured for 6 h in conditions mimicking hypoxia expressed lower amounts of HIF-1alpha mRNA. In two rat cell lines, aHIF transcript levels were greatly augmented after a 6-h incubation in these conditions, whereas in a mouse cell line, aHIF level was significantly reduced. PMID- 15363853 TI - Emergence of Talanin protein associated with human uric acid nephrolithiasis in the Hominidae lineage. AB - Recently, we identified a susceptibility locus for human uric acid nephrolithiasis (UAN) on 10q21-q22 and demonstrated that a novel gene (ZNF365) included in this region produces through alternative splicing several transcripts coding for four protein isoforms. Mutation analysis showed that one of them (Talanin) is associated with UAN. We examined the evolutionary conservation of ZNF365 gene through a comparative genomic approach. Searching for mouse homologs of ZNF365 transcripts, we identified a highly conserved mouse ortholog of ZNF365A transcript, expressed specifically in brain. We did not found a mouse homolog for ZNF365D transcript encoding the Talanin protein, even if we were able to identify the corresponding genomic region in mouse and rat not yet organized in canonical gene structure suggesting that ZNF365D was originated after the branching of hominoid from rodent lineage. In mouse and in most mammals, a functional uricase degrades the uric acid to allantoin, but uricase activity was lost during the Miocene epoch in hominoids. Searching for the presence of Talanin in Primates, we found a canonical intron-exon structure with several stop codons preventing protein production in Old World and New World monkeys. In humans, we observe expression and we have evidence that ZNF365D transcript produces a functional protein. It seems therefore that ZNF365D transcript emerged during primate evolution from a noncoding genomic sequence that evolved in a standard gene structure and assumed its role in parallel with the disappearance of uricase, probably against a disadvantageous excessive hyperuricemia. PMID- 15363854 TI - Truncation and mutagenesis analysis of the human X-arrestin gene promoter. AB - X-arrestin (arrestin-3) is an arrestin present specifically in the outer segments of red-, green-, and blue-cone photoreceptors. The X-arrestin gene is on Xcen q22, and consists of 17 exons with a promoter containing a TATA box and elements important for photoreceptor expression, including three CRX and one PCE-1-like element. In order to delineate the promoter structure necessary for the pan-cone specific expression of X-arrestin, the expression of the gene in retinoblastoma cell lines was investigated, and a structure-function analysis of the promoter was conducted in the appropriate cellular substrate. Expression of X-arrestin was detected at a low level in the Y79 retinoblastoma cell line but not in the WERI retinoblastoma cell line. Truncation and expression analysis of the X-arrestin promoter in Y79 showed maximal activity in the proximal 378-bp region containing the CRX and PCE-1-like elements upstream of the TATA and CAAT boxes and a negative regulator in the distal 1-2-kbp region. Mutagenesis of the three CRX and PCE-1-like elements and expression analysis demonstrated complete elimination of the promoter activity. Mutagenesis of the TATA box and PCE-1-like element individually resulted in similar decrease in promoter activity, but the decrease in the promoter activity was greater when the CRX elements were mutagenized with a 5' to 3' spatial gradient in the negative effect, suggesting a cooperative effect of the three CRX elements. The regulation of expression from this promoter may involve the binding of a multi-protein enhanceosome complex at the CRX triplet and the PCE-1-like element, resulting in the recruitment and activation of the RNA polymerase II complex at the downstream TATA box. PMID- 15363855 TI - The origin and evolution of eucaryal HIS7 genes: from metabolon to bifunctional proteins? AB - The fifth step of histidine biosynthesis is catalysed by an imidazole glycerol phosphate (IGP) synthase. In Archaea and Bacteria, the active form of IGP synthase is a stable 1:1 dimeric complex constituted by a glutamine amidotransferase (GAT) and a cyclase, the products of hisH and hisF. In Eucarya, the two activities are associated with a single bifunctional polypeptide encoded by HIS7. In this work, we report a comparative analysis of the amino acid sequence of all the available HisH, HisF and HIS7 proteins, which allowed depicting a likely evolutionary pathway leading to the present-day bifunctional HIS7 genes. According to the model that we propose, the bifunctional HIS7 gene is the outcome of a gene fusion event between two independent ancestral cistrons encoding an amidotransferase and a cyclase, respectively. The phylogenetic distribution of the eucaryal HIS7 genes and the analysis of all the available prokaryotic counterparts (hisH and hisF) revealed the absence of such fusions in prokaryotes, suggesting that the fusion event very likely occurred in an early stage of eucaryal evolution and was fixed in the nucleated cells. The biological significance of this gene fusion is also discussed. PMID- 15363856 TI - Structure and expression of the gene encoding a Broad-Complex homolog in the silkworm, Bombyx mori. AB - The steroid hormone ecdysone (20-hydroxyexdysone) initiates metamorphosis and also larval ecdysis in many insects by activating a cascade of genes that includes primary response genes (early genes), most of which encode transcriptional regulators, and secondary response genes (late genes) regulated by the early genes. One of the early genes, Broad-Complex (BR-C), a key regulator of the ecdysone cascade, shares a common amino-terminal BTB domain which is fused by alternative splicing to one of four pairs of C(2)H(2) zinc finger domains (Z1, Z2, Z3, and Z4). cDNAs for BR-C (BmBR-C) were isolated from the silkworm Bombyx mori. These genes showed 90.3% and 98.2% amino acid identity with the Drosophila BR-C and Manduca BR-C in the N-terminal BTB domain; 96.0%, 90.7%, and 85.2% identity with the three zinc finger domains of the Drosophila Z1, Z2, and Z4 isoforms; and 96.3% and 98.1% identity with the two zinc finger domains of the Manduca Z2 and Z4 isoforms, respectively. Partial genomic sequencing (from the 3' region of the core sequence to the 3' region of the Z3 class zinc finger-coding sequence) of the BmBR-C gene showed that four exons coding the zinc finger domains are arranged the same as the BR-C gene in Drosophila. The amino acid sequence predicted from the genomic sequence corresponding to the BmBR-C Z3 class zinc finger domain is 100% identical to the Z3 isoforms of Drosophila and Manduca. We examined expression patterns of the BmBR-C isoforms during late larval to pupal development in the epidermis, fatbody and silk gland. During the metamorphic transformation, the epidermis and silk gland are completely histolyzed; however, the fat body survives into the adult phase. Expression patterns of BmBR-C during development differed extensively between the histolyzed group and the survival group. The BmBR-C expression patterns in silk glands also differed between the anterior and other areas (the middle and posterior silk glands). PMID- 15363857 TI - Genomic organizations of two small subunit ADP-glucose pyrophosphorylase genes from sweetpotato. AB - The genomic features of the small subunit ADP-glucose pyrophosphorylase (AGPase) isoforms are different in barley and maize. The two isoforms found in barley originated from one single gene through alternative splicing, while two independent genes encode the two isoforms in maize. To ascertain the genomic organizations of two small subunit AGPase isoforms in sweetpotato (ibAGP1 and ibAGP2), we isolated genomic DNAs containing the entire coding regions of two genes. Complete genomic structures of ibAGP1 and ibAGP2 were ascertained by the sequencing of these genomic regions. The transcribed regions of ibAGP1 and ibAGP2, comprising nine exons and eight introns, were distributed over 3.9 and 4.0 kb, respectively. The eight introns differed in length, from 76 to 946 bp in ibAGP1, and from 76 to 811 bp in ibAGP2, while the locations of introns in ibAGP1 and ibAGP2 were identical. There was 46-58% sequence identity between the intron sequences of the two genes. Intron sequence analyses suggested that either duplication in each intron, or gene conversion between introns of two isoforms, might cause major intron size differences between the two genes. Altogether, these results indicate that two small subunit AGPase isoforms in sweetpotato are encoded by two independent genes, in a fashion similar to that of maize small subunit AGPase genes. PMID- 15363858 TI - The molecular phylogeny of oysters based on a satellite DNA related to transposons. AB - We have analysed a centromeric satellite DNA family that is conserved in several commercial and non-commercial oyster species (Ostrea edulis, O. stentina, Crassostrea angulata, C. gigas, C. gasar, C. ariakensis, C. virginica and C. sikamea). This satellite DNA family is composed of AT-rich repeat sequences of 166+/-2 bp and presents a 9-bp motif similar to the mammalian CENP-B box. The homology of oyster HindIII satellite DNA with satellite DNAs from other bivalves and its relation to a part of a mobile element suggest the existence of an ancient transposable element as a generating unit of satellite DNA in bivalve molluscs. Taking advantage of its degree of conservation in oyster species, we have used this element as a taxonomic marker. This marker clearly supports a high degree of differentiation between O. edulis and O. stentina, and, conversely, upholds the contention that C. gigas and C. angulata are the same species. Finally, we have used HindIII satellite DNA as a phylogenetic marker between these species, revealing two clades, one formed by Asiatic species (C. angulata, C. gigas and C. ariakensis) and another by the European, American and African species (O. edulis, C. virginica and C. gasar, respectively). PMID- 15363859 TI - A new pair of CR1-like LINE and tRNA-derived SINE elements in Podarcis sicula genome. AB - We have identified and characterized a new pair of LINE and SINE elements, called Lucy-1 CR1-like LINE and P.s.1/SINE, respectively, in Podarcis sicula genome. The 3'-tail region in the 3' untranslated region (UTR) of Lucy-1 element is almost identical to the of P.s.1/SINE element. This identity suggests that the P.s.1/SINE element, during evolution, has gained the 3'-end sequence of the Lucy 1 element and has exclusively recruited the enzymatic machinery of its partner CR1 LINE for retroposition. Moreover, the complex molecular organization around Lucy-1 insertion site is discussed and we found that Lucy-1 insertion is associated with the calcium binding transporter gene. Our results confirm that the retrotransposons can be an additional source of genomic diversification and the evolution of the retrotransposable elements can be a vector force shaping genomes by reassorting DNA domains thus forming a new DNA arrangement. PMID- 15363860 TI - RANKL/RANK/OPG: new therapeutic targets in bone tumours and associated osteolysis. AB - The emergence of the molecular triad osteoprotegerin (OPG)/Receptor Activator of NF-kB (RANK)/RANK Ligand (RANKL) has helped elucidate a key signalling pathway between stromal cells and osteoclasts. The interaction between RANK and RANKL plays a critical role in promoting osteoclast differentiation and activation leading to bone resorption. OPG is a soluble decoy receptor for RANKL that blocks osteoclast formation by inhibiting RANKL binding to RANK. The OPG/RANK/RANKL system has been shown to be abnormally regulated in several malignant osteolytic pathologies such as multiple myeloma [MM, where enhanced RANKL expression (directly by tumour cells or indirectly by stromal bone cells or T-lymphocytes)] plays an important role in associated bone destruction. By contrast, production of its endogenous counteracting decoy receptor OPG is either inhibited or too low to compensate for the increase in RANKL production. Therefore, targeting the OPG/RANK/RANKL axis may offer a novel therapeutic approach to malignant osteolytic pathologies. In animal models, OPG or soluble RANK was shown both to control hypercalcaemia of malignancy and the establishment and progression of osteolytic metastases caused by various malignant tumours. To this day, only one phase I study has been performed using a recombinant OPG construct that suppressed bone resorption in patients with multiple myeloma or breast carcinoma with radiologically confirmed bone lesions. RANK-Fc also exhibits promising therapeutic effects, as revealed in animal models of prostate cancer and multiple myeloma. If the animal results translate to similar clinical benefits in humans, using RANK-Fc or OPG may yield novel and potent strategies for treating patients with established or imminent malignant bone diseases and where standard therapeutic regimens have failed. PMID- 15363861 TI - Intracellular signaling mechanisms in photodynamic therapy. AB - In photodynamic therapy (PDT) a sensitizer, light and oxygen are used to induce death of tumor cells and in the treatment of certain noncancerous conditions. Cell death in PDT may occur by apoptosis or by necrosis, depending on the sensitizer, on the PDT dose and on the cell genotype. Some sensitizers that have been used in PDT are accumulated in the mitochondria, and this may explain their efficiency in inducing apoptotic cell death, both in vitro and in vivo. In this review we will focus on the events that characterize apoptotic death in PDT and on the intracellular signaling events that are set in motion in photosensitized cells. Activation of phospholipases, changes in ceramide metabolism, a rise in the cytosolic free Ca2+ concentration, stimulation of nitric oxide synthase (NOS), changes in protein phosphorylation and alterations in the activity of transcription factors and on gene expression have all been observed in PDT treated cells. Although many of these metabolic reactions contribute to the demise process, some of them may antagonize cell death. Understanding the signaling mechanisms in PDT may provide means to modulate the PDT effects at the molecular level and potentiate its antitumor effectiveness. PMID- 15363862 TI - DNA methylation in prostate cancer. AB - Prostate cancer is the most common malignancy and the second leading cause of cancer death among men in the United States. There are three well-established risk factors for prostate cancer: age, race and family history. The molecular bases for these risk factors are unclear; however, they may be influenced by epigenetic events. Epigenetic events covalently modify chromatin and alter gene expression. Methylation of cytosine residues within CpG islands on gene promoters is a primary epigenetic event that acts to suppress gene expression. In tumorigenesis, the normal functioning of the epigenetic-regulatory system is disrupted leading to inappropriate CpG island hypermethylation and aberrant expression of a battery of genes involved in critical cellular processes. Cancer dependent epigenetic regulation of genes involved in DNA damage repair, hormone response, cell cycle control and tumor-cell adhesion/metastasis can contribute significantly to tumor initiation, progression and metastasis and, thereby, increase prostate cancer susceptibility and risk. In this review, we will discuss current research on genes that are hypermethylated in human prostate cancer. We will also discuss the potential involvement of DNA methylation in age-related, race-related and hereditary prostate cancer, and the potential use of hypermethylated genes as biomarkers to detect prostate cancer and assess its risk. PMID- 15363863 TI - Depression in tension-type headache sufferers: bystander or villain? PMID- 15363864 TI - Do we need a communal coping model of pain catastrophizing? An alternative explanation. PMID- 15363865 TI - Depression increases onset of tension-type headache following laboratory stress. AB - The aim of this study is to examine the influence of depression on headache onset following laboratory stress and on psychophysiological variables associated with tension-type headaches (TTHs). Diagnostic interviews identified three groups: headache prone and depressed (HP/D, N = 13); headache prone not depressed (HP/ND, N = 22); and healthy controls (HC, N = 13). Ss completed a laboratory stress task. Blind evaluations of pericranial muscle tenderness (PMT) and pressure pain thresholds (PPT) were obtained immediately before, immediately after and 24 h post-task. Ss also recorded headaches (HA) before, during, immediately post-task and for 24 h post-task. HP/D Ss were more likely than HP/ND Ss or HC Ss to report a headache during and immediately following the stress task (P < 0.05). HP/D Ss exhibited higher PMT than HP/ND Ss or HC Ss before and following the stress task (P < 0.05). HP/D Ss exhibited lower finger PPT at all assessments and lower temporalis PPT at two of three assessments than HC Ss (P < 0.05) but did not significantly differ from HP/ND Ss at most assessments. Depression increased vulnerability to TTH following laboratory stress and was associated with elevated PMT. In individuals with frequent headaches, depression may aggravate existing central sensitization increasing vulnerability to TTHs. PMID- 15363866 TI - Analgesia following surgery in children with and without cognitive impairment. AB - Both children and adults with cognitive impairment (CI) have historically been excluded from research examining pain. This is unfortunate since patients with CI may be at higher risk for experiencing pain or having their pain undertreated due to the difficulty of pain assessment and communication. There are now several published reports about the general pain experience of both adult and pediatric patients with cognitive impairment. The purpose of this study was to compare the amount and type of pain medication administered in children with and without CI after surgery to ascertain if there were any differences in analgesic administration patterns between these two groups. One hundred and fifty-two children with borderline to profound CI and 138 non impaired (NI) children were recruited to participate. Analgesic administration data include type and amount of opioid, type of non-opioid medication, and prescribed discharge medications. Results of this study show that children with CI undergoing surgery received less opioid in the perioperative period than children without CI. However, children with CI received comparable amounts and types of analgesics in the postoperative period as children without CI. PMID- 15363867 TI - Self-efficacy, fear avoidance, and pain intensity as predictors of disability in subacute and chronic musculoskeletal pain patients in primary health care. AB - This study examined the relations between disability, as measured by the Pain Disability Index (PDI) and self-efficacy, fear avoidance variables (kinesiophobia and catastrophizing), and pain intensity, using a prospective design. Two primary health care samples (n(1)=210; n(2)=161) of patients with subacute, chronic or recurring musculoskeletal pain completed sets of questionnaires at the beginning of a physiotherapy treatment period. Multiple hierarchial regression analyses showed that self-efficacy explained a considerably larger proportion of the variance in disability scores than the fear avoidance variables in the first sample. This finding was replicated in the second sample. Pain intensity explained a small, but significant proportion of the variance in disability scores in one sample only. Gender, age, and pain duration were not related to disability. These findings suggest that self-efficacy beliefs are more important determinants of disability than fear avoidance beliefs in primary health care patients with musculoskeletal pain. The findings also suggest that pain-related beliefs, such as self-efficacy and fear avoidance, in turn, are more important determinants of disability than pain intensity and pain duration in these patients. PMID- 15363868 TI - The anti-hyperalgesic effects of carbamazepine and oxcarbazepine are attenuated by treatment with adenosine receptor antagonists. AB - The antinociceptive effects of carbamazepine and oxcarbazepine, and the influence of caffeine, were examined in a paw pressure test in rats. Carbamazepine (10-40 mg/kg; intraperitoneal, i.p.) and oxcarbazepine (40-160 mg/kg; i.p.) caused a significant dose-dependent reduction of the paw inflammatory hyperalgesia induced by concanavalin A (Con A), intraplantarly (i.p1.). A comparable pattern of antinociceptive effect of carbamazepine and oxcarbazepine was observed; the only difference is their potency, in that carbamazepine was about three times more potent than oxcarbazepine. Caffeine (5-20mg/kg; i.p.), a non-selective adenosine receptor antagonist, significantly depressed the antinociceptive effects of carbamazepine and oxcarbazepine, in a dose- and time-dependent manner. Also, a significant depression of the antinociceptive effects of carbamazepine and oxcarbazepine was observed by pretreatment with 1,3-dipropyl-8-cyclopentylxantine (DPCPX, 0.4 and 0.8 mg/kg; i.p.), an adenosine A(1) receptor antagonist. These findings indicate that, in a paw inflammatory hyperalgesia in rats, the antinociceptive effects of both drugs are, at least partially, mediated by adenosine A(1) receptors. In conclusion, the present study suggests the potential clinical importance of carbamazepine and oxcarbazepine in the treatment of inflammatory pain. In addition, caffeine consumption could possibly depress the analgesic effects of both anticonvulsive drugs. PMID- 15363869 TI - The effect of triamcinolone hexacetonide on the spontaneous and mechanically induced ectopic discharge following lingual nerve injury in the ferret. AB - Investigations into the aetiology of nerve injury-induced dysaesthesia have revealed the development of spontaneous and mechanically-induced activity from damaged axons. Pharmacological manipulation of this activity could provide a method of treatment for this intractable condition. This study has investigated the effect of a corticosteroid applied to the injury site, as these agents are known to reduce inflammation and scarring. In 24 anaesthetised adult ferrets the left lingual nerve was sectioned and the animals allowed to recover. In eight of these animals the nerve was re-exposed under anaesthesia after 1 month and 100 microl of corticosteroid (triamcinolone hexacetonide, 20 mg/ml) was injected into and around the injury site. In eight others, 100 microl of the steroid carrier was injected, and the eight remaining animals were used as controls. In terminal experiments under general anaesthesia, 3 months after the initial injury, electrophysiological recordings were made from axons in fine filaments dissected from the nerve central to both the injury site and junction with the chorda tympani nerve. Spontaneous activity (SA) was found in approximately 13% of units in control animals, 12% following the application of steroid, and 14% in the carrier group. Mechanically-induced activity at the injury site was found in approximately 13% of units in controls, significantly fewer after the application of steroid 4% (P<0.001) and 12% in the carrier group. These data suggest that local application of the corticosteroid triamcinolone hexacetonide could reduce the level of mechanically-induced, but not spontaneous, dysaesthesia following lingual nerve injury. PMID- 15363870 TI - Can one predict the likely specific orofacial pain syndrome from a self-completed questionnaire? AB - To estimate the prevalence of orofacial pain (OFP) by specific diagnostic subgroups in the general population. Cross-sectional population study. General medical practice in South East Cheshire, UK. Participants of baseline investigation who completed the full postal questionnaire (1510, adjusted study participation rate 81%). Clinical examination was attended by 126 (43%) of all the participants who reported OFP in the questionnaire. These individuals were classified as musculoligamentous/soft tissue type, dentoalveolar or neurological/vascular. OFP duration, location, descriptors and statements on OFP were predictors of classification group. The estimated prevalence in the general population of musculoligamentous/soft tissue type of OFP was 7%, dentoalveolar 7% and neurological/vascular 6%. This study has derived a statistical model to classify participants with OFP into three broad groups (musculoligamentous/soft tissue, dentoalveolar and neurological/vascular) based on questionnaire information about OFP (OFP chronicity, location and verbal descriptors of pain). It is potentially useful in large population studies of OFP, where a clinical examination is not possible, however, further validation of its performance in large populations are necessary. PMID- 15363871 TI - Peripheral and central p38 MAPK mediates capsaicin-induced hyperalgesia. AB - The stress-activated mitogen-activated protein kinase (MAPK) p38 is emerging as an important mediator of pain. The present study examined the possible involvement of peripheral and spinal p38 MAPK in capsaicin-induced thermal hyperalgesia. Topical capsaicin produced phosphorylation of p38 MAPK in the skin from the affected hindpaw as well as the corresponding lumbar spinal cord in a time dependent manner. Topical capsaicin produced robust C-fiber mediated thermal hyperalgesia that was inhibited by systemic, local peripheral, or central intrathecal pre-treatment with the p38 MAPK inhibitor, SD-282. Intraperitoneal SD 282 (10-60 mg/kg) significantly and dose-dependently attenuated capsaicin-induced C-fiber mediated thermal hyperalgesia. Similarly, 0.1-5mg/kg subcutaneous SD-282 in the hindpaw dose-dependently attenuated capsaicin-induced thermal hyperalgesia. Intrathecal administration of 1microg SD-282 was also anti hyperalgesic in this model. Functionally, SD-282 decreased capsaicin-induced release of calcitonin gene related peptide in an in vitro skin release assay, consistent with a role for p38 MAPK in peripheral nerve function. These results suggest that p38 MAPK plays a role in the development of hyperalgesic states, exerting effects both centrally in the spinal cord and peripherally in sensory C fibers. PMID- 15363872 TI - Efficacy and safety of valdecoxib for treatment of osteoarthritis and rheumatoid arthritis: systematic review of randomised controlled trials. AB - Our objective was to determine the efficacy and safety of valdecoxib (a cyclo oxygenase 2 inhibitor) in the treatment of arthritis. Randomised, controlled trials comparing 10 or 20mg valdecoxib with placebo or non-steroidal anti inflammatory drugs (NSAIDs) in patients with active osteoarthritis or rheumatoid arthritis. The manufacturer provided clinical trial reports. Data were combined through meta-analysis. Main outcomes were patient global rating of arthritis, arthritis pain, Western Ontario and McMaster Universities indices for osteoarthritis, American College of Rheumatology indices for rheumatoid arthritis, discontinuation, endoscopic ulcers, clinically significant upper gastrointestinal or renal events. Nine trials (five in osteoarthritis, four in rheumatoid arthritis) were included with 5726 patients. Overall, valdecoxib 10 and 20mg were superior to placebo and equivalent in efficacy to maximum daily doses of NSAIDs. Significantly fewer discontinuations because of gastrointestinal adverse events (4% versus 8%), or endoscopic ulcers of 3mm or more (5% versus 13%) occurred with valdecoxib compared with NSAIDs. Clinically significant upper gastrointestinal events occurred in 2/2733 (0.1%) with valdecoxib compared with 8/1846 (0.4%) with NSAIDs. Rates of clinically significant renal events were the same (2-3%) for valdecoxib and NSAIDs. At an appropriate dose valdecoxib was as effective as NSAIDs in osteoarthritis and rheumatoid arthritis. There were fewer gastrointestinal adverse event withdrawals and endoscopically detected ulcers. Convincing evidence of reduced major gastrointestinal adverse events could not be addressed by the trials. PMID- 15363873 TI - Dietary fat and protein interact in suppressing neuropathic pain-related disorders following a partial sciatic ligation injury in rats. AB - Chronic neuropathic sensory disorders (CNSD) of rats receiving a partial sciatic nerve ligation injury (the PSL model) are suppressed by dietary soy protein. Although previously shown to modify nociceptive behavior in acute pain models, dietary fat has never been tested for its putative analgesic properties in chronic pain states. Here we tested the role of dietary fat, protein and fat/protein interactions in the development of tactile allodynia and heat hyperalgesia in PSL-injured rats. Male Wistar rats were fed nine different diets, comprising of three proteins (soy, casein and albumin) and three fats (corn, soy and canola) for a week preceding PSL injury and for 2 weeks thereafter. Rats' responses to tactile and noxious heat stimuli were tested before surgery and 3, 7 and 14 days afterwards. Tactile and heat sensory abnormalities following PSL injury were significantly different among the nine dietary groups. Consumption of corn and soy fats suppressed the levels of tactile and heat allodynia and hyperalgesia, whereas consumption of soy and casein proteins was associated with lower levels of heat hyperalgesia but not tactile allodynia. A significant fat/protein interaction was found for the heat but not tactile stimuli. We conclude that dietary fat is a significant independent predictor of levels of neuropathic sensory disorders in rats and that this effect is accentuated by dietary protein. The mechanisms by which fat suppresses neuropathic disorders have yet to be determined. PMID- 15363874 TI - Oral ibandronate improves bone pain and preserves quality of life in patients with skeletal metastases due to breast cancer. AB - The objective of this study is to assess the effect of oral ibandronate on bone pain and quality of life in women with metastatic bone disease from breast cancer. In two double-blind, placebo-controlled studies, 564 patients were randomised to receive oral ibandronate, 50mg once daily, or placebo for up to 96 weeks. Throughout the studies, we assessed bone pain (on a 5-point scale from 0=none to 4=intolerable), analgesic use (7-point scale) and quality of life (European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire-C30 [EORTC QLQ-C30], 100-point scale). Oral ibandronate significantly reduced and maintained bone-pain scores below baseline throughout the 96-week study period (at endpoint, -0.1 vs +0.2, P=0.001 vs placebo). Analgesic use increased in both groups; however, the increase was significantly less in the ibandronate group (0.60 vs 0.85, P=0.019). Although quality of life deteriorated during the study, the decrease in quality of life was significantly lower with ibandronate therapy (-8.3 vs -26.8, P=0.032). Drug-related adverse events were generally minor and as expected with oral bisphosphonates. Oral ibandronate had beneficial effects on bone pain and quality of life and was well tolerated. These results suggest that this treatment is of considerable clinical value as a co-analgesic to patients with painful bone metastases. PMID- 15363875 TI - Clinical tolerability of perioperative tenoxicam in 1001 patients--a prospective, controlled, double-blind, multi-centre study. AB - We investigated adverse events (AEs) associated with perioperative tenoxicam in a double-blind, prospective, randomised study. Patients undergoing surgery, screened for contraindications to non-steroidal anti-inflammatory drug, received tenoxicam (n=750) on 2843 days or placebo (n=251) on 988 days, in courses of 1-12 days. There was no increase in the overall incidence of side effects with tenoxicam (33 vs 38% with placebo: P=0.15), or in major side effects (3.9 vs 2.0% with placebo: P=0.11). Of major side effects possibly or probably related to tenoxicam (2.1 vs 1.2% with placebo: P=0.26), all but one involved post-operative surgical site bleeding. However, in the subgroup of patients undergoing otorhinolaryngology surgery, surgical site bleeding occurred in 18 of 171 (10.5%) patients on tenoxicam and one of 57 (1.8%) on placebo (P=0.026); of these, nine in the tenoxicam group and 0 in the placebo were classified as major (P=0.07). One patient on tenoxicam experienced endoscopically proven duodenal ulceration with malaena. In conclusion, perioperative tenoxicam is well tolerated in comparison with placebo and the incidence of drug-related major AEs (other than post-operative bleeding) is no greater than 1 in 150 in low risk patients, but in patients undergoing otorhinolaryngological surgery there may be an increased risk of post-operative bleeding. PMID- 15363876 TI - Alteration of the second branch of the trigeminal nerve activity following inferior alveolar nerve transection in rats. AB - After transection of the inferior alveolar nerve (IAN), the whisker pad area, which is innervated by the infraorbital nerve (ION) that was not injured, showed hypersensitivity to mechanical stimulation. Two days after IAN transection, threshold intensity for escape behavior to mechanical stimulation of the ipsilateral whisker pad area was less than 4.0 g, indicating mechanical allodynia. A total of 68 single fiber discharges were recorded from ION fibers at 3 days after IAN transection. The responses of C- and A-fibers were classified according to their conduction velocity. The C-fiber activities were not affected by IAN transection, whereas A-fiber activities were significantly enhanced by IAN transection as indicated by an increase in background activity and mechanically evoked response. Since the A-fiber responses were significantly affected by IAN transection, patch clamp recording was performed from middle to large diameter retrogradely labeled and acutely dissociated trigeminal ganglion (TRG) neurons. The I(K) (sustained) and I(A) (transient) currents were significantly smaller and hyperpolarization-activated current (I(h)) was significantly larger in TRG neurons of rats with IAN transection as compared to those of naive rats. Furthermore, current injection into TRG neurons induced high frequency spike discharges in rats with IAN transection. These data suggest that changes in K(+) current and I(h) observed in the uninjured TRG neurons reflect an increase in excitability of TRG neurons innervated by the ION after IAN transection, resulting in the development of mechano-allodynia in the area adjacent to the injured IAN innervated region. PMID- 15363877 TI - Catastrophizing as a mediator of sex differences in pain: differential effects for daily pain versus laboratory-induced pain. AB - Sex differences in the experience of pain have been widely reported, with females generally reporting more frequent clinical pain and demonstrating greater pain sensitivity. However, the mechanisms underpinning such differences, while subject to intense speculation, are not well-characterized. Catastrophizing is a cognitive and affective process that relates strongly to enhanced reports of pain and that varies as a function of sex. It is thus a prime candidate to explain sex differences; indeed, several prior studies offer evidence that controlling for catastrophizing eliminates the gap between men and women in reported pain. We recruited 198 healthy young adults (115 female) who took part in laboratory studies of pain responses, including thermal pain, cold pain, and ischemic pain, and who also completed questionnaires assessing catastrophizing, mood, and day-to day painful symptoms (e.g. headache, backache). Women reported greater levels of catastrophizing, more recent painful symptoms, and demonstrated lower pain thresholds and tolerances for noxious heat and cold relative to men. Mediational analyses suggested that after controlling for negative mood, catastrophizing mediated the sex difference in recent daily pain but did not mediate the much larger sex differences in pain threshold and tolerance. These findings highlight the role of catastrophizing in shaping pain responses, as well as illuminating potentially important differences between experimental pain assessment and the clinical experience of pain. PMID- 15363878 TI - Spatial summation of heat pain within and across dermatomes in fibromyalgia patients and pain-free subjects. AB - The mechanisms of spatial summation of pain (SSP) include pain coding dependent on impulse frequency and the number of recruited central neurons. However, SSP may also be influenced by pain inhibitory mechanisms, such as diffuse noxious inhibitory controls. Abnormal interactions between pain inhibitory mechanisms and SSP may be relevant for chronic pain conditions such as fibromyalgia (FM) and may help explain why widespread pain is characteristic for this chronic pain syndrome. The present study was designed to determine the difference of thermal SSP in the upper extremities between FM and normal control (NC) subjects, particularly within and across dermatomes of the hand. Fourteen NC and 19 FM subjects were enrolled in this study. SSP testing sessions involved immersion of each individual fingertip as well as stepwise immersion of the fingers, hands, and forearms in a hot water bath (46 degrees Celsius) for 5s and 20s. In addition, immersion of several fingertips across dermatome C(7)-C(8) was compared to progressive immersion of the index finger (dermatome C(7)). These experiments demonstrated significant spatial summation of heat-induced pain in both FM and NC subjects. SSP was most extensive within the fingers, and became negligible as the stimulus area increased above the hand. Furthermore, SSP was more pronounced within one dermatome such as that of the index finger than across several dermatomes of the hand. These results were similar for both FM and NC subjects. Thus, mechanisms of SSP, including possible inhibitory factors that limit this relevant pain mechanism, appear to be similar for both FM and NC subjects. PMID- 15363879 TI - Development of tactile allodynia and thermal hyperalgesia by intrathecally administered platelet-activating factor in mice. AB - Platelet-activating factor (PAF) is a potent inflammatory lipid mediator in peripheral tissues. However, its role in mediation of nociception in central nervous system is unknown. In the present study, whether PAF plays some role in pain transduction in the spinal cord was studied in mice. Intrathecal injection of PAF induced tactile pain, tactile allodynia at as low as 10 fg to 1 pg with a peak response at 100 fg, while lyso-PAF was without effect in the range of doses. Tactile allodynia induced by PAF was blocked by a PAF receptor antagonists, TCV 309, WEB 2086 and BN 50739. The expression of PAF receptor mRNA by RT-PCR was observed in DRG and spinal cord in mice. ATP P2X receptor antagonists, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid and 2',3'-O-(2,4,6 trinitrophenyl)adenosine 5-triphosphate, NMDA receptor antagonist, MK 801 and nitric oxide synthetase inhibitor, 7-nitroindazole blocked the PAF-induced tactile allodynia. PAF-induced tactile allodynia and thermal hyperalgesia disappeared in neonatally capsaicin-treated adult mice, while tactile allodynia but not thermal hyperalgesia induced by intrathecally injected alpha,beta methylene ATP, a P2X receptor agonist, was capsaicin-insensitive. The present study demonstrated that PAF is a potent inducer of tactile allodynia and thermal hyperalgesia at the level of the spinal cord. PAF-evoked tactile allodynia is suggested to be mediated by ATP and the following NMDA and NO cascade through capsaicin-sensitive fiber, different from exogenously injected alpha,beta methylene ATP which is insensitive to capsaicin treatment. PMID- 15363880 TI - Reduced heat sensitivity and epidermal nerve fiber immunostaining following single applications of a high-concentration capsaicin patch. AB - Capsaicin-containing plant extracts have been used as topical treatments for a variety of pain syndromes for many centuries. Current products containing capsaicin in low concentrations (usually 0.025-0.075% w/w) have shown efficacy against a variety of pain conditions in clinical studies. However, in order to produce significant analgesic effects, these formulations require frequent re dosing, often as much as three to five times daily for several weeks. Previous functional and immunohistochemical studies following prolonged exposures to low concentration capsaicin cream suggested that the duration and onset of analgesic efficacy correlate with a reduction of cutaneous nociceptive sensory nerve fiber responsiveness and immunostaining. The purpose of the present study was to determine whether a single topical application of a high-concentration capsaicin containing (8%w/w) patch for 120 min or less would induce similar effects on cutaneous nociceptive nerve fibers. Seven days following patch application, changes in heat and cold perception thresholds were determined by quantitative sensory testing and punch biopsies were collected to assess epidermal nerve fiber (ENF) immunostaining density at the application site using PGP 9.5 as a marker. The results show a significant reduction of heat, but not cold, sensitivity and reduction of ENF immunostaining with high-capsaicin concentration patch applications for 60 or 120 min, compared to placebo patch applications. Application sites exposed to low-capsaicin concentration (0.04%w/w) patches for 120 min or high-concentration patches for 30 min were not significantly different from placebo with respect to either thermal threshold detection or ENF immunostaining. The ability of a single 60 min high-concentration patch application to mimic effects produced by prolonged exposure to low-concentration capsaicin creams suggests a new approach to the management of chronic pain syndromes. PMID- 15363881 TI - Controlled trial of Internet-based treatment with telephone support for chronic back pain. AB - The purpose of this study was to investigate the effects of an Internet-based cognitive-behavioral intervention with telephone support for chronic back pain. Participants who met the criteria for chronic back pain (N=56) were randomly assigned to either an Internet-based cognitive behavioral self-help treatment or to a waiting-list control condition. The study period lasted 8 weeks and consisted of 1 week of self-monitoring prior to the intervention, 6 weeks of intervention, and 1 week of post-intervention assessment. Treatment consisted of education, cognitive skill acquisition, behavioral rehearsal, generalization and maintenance. The dropout rate was 9% (N=5). Results showed statistically significant improvements in catastrophizing, control over pain and ability to decrease pain. Some improvement was found in both the control group and the treatment group. A follow-up of 3 months after treatment termination was completed in 92% (N=47) of the participants who completed the treatment intervention. Follow-up results showed that some improvement was maintained. Findings indicate that Internet-based self-help with telephone support, based on established psychological treatment methods, holds promise as an effective approach for treating disability in association with pain. PMID- 15363882 TI - Abnormal contralateral pain responses from an intradermal injection of phenylephrine in a subset of patients with complex regional pain syndrome (CRPS). AB - We have examined the effect of an intradermal injection of phenylephrine (1mg/0.1 ml), an alpha-1-adrenoceptor agonist in normal subjects, and patients with sympathetically-independent (SIP) and sympathetically-maintained pain (SMP). Normal subjects and SIP patients experienced only brief stinging pain, while subsets of both sympathectomized and non-sympathectomized SMP patients (6/9 and 4/8, respectively) experienced an additional abnormal pain response accompanied by mechano-allodynia around the injection site. Both the normal and abnormal pain response after intradermal phenylephrine are similar to those observed with intradermal norepinephrine. In contrast to previous reports in the literature, we found that three sympathectomized SMP patients (who, however, had failed to experience pain relief after surgical sympathectomy despite very good relief after sympathetic blocks) also experienced abnormal pain and mechano-allodynia when phenylephrine was injected to a limb contralateral to the symptomatic sympathectomized extremity. Abnormal pain response evoked by norepinephrine or phenylephrine injection in the ipsilateral symptomatic limb of SMP patients may be due to injury-evoked nociceptor responsiveness to catecholamines. However, such a response in contralateral asymptomatic limbs suggests an additional factor that more likely than not is of central origin and may or may not be related to sympathectomy and its success or failure to treat pain. PMID- 15363883 TI - Topical clonidine antinociception. AB - Clonidine, an alpha-2 adrenergic agonist, is an extremely potent antinociceptive agent. However, the therapeutic utility of systemic clonidine for the treatment of pain is limited by centrally mediated side effects including sedation, hypotension and rebound hypertension. Given that alpha-2 adrenoceptors are expressed on the peripheral and central terminals of nociceptive fibers, we administered clonidine topically in order to avoid central effects. Here, we demonstrate that topical administration of clonidine to mice (via tail immersion) elicited antinociception in the radiant heat tail-flick test. The magnitude of antinociception was dependent upon the duration of exposure to the clonidine solution. Further, the antinociceptive activity of clonidine was limited to the portion of the tail exposed to drug solution suggesting that the actions of clonidine were locally mediated. Systemic pretreatment with the alpha-2 receptor antagonist, yohimbine, blocked the antinociceptive activity of topical clonidine. Concentrations of clonidine administered locally that were antinociceptive did not impair motor coordination as measured by the rota-rod test. However, doses of clonidine administered systemically that produced antinociception significantly impaired motor coordination. Repeated daily topical administration of clonidine resulted in antinociceptive tolerance. Tolerance to the antinociceptive actions of clonidine was not blocked by topical administration of the NMDA antagonist, ketamine. In conclusion, topical administration of clonidine elicits antinociception by blocking the emerging pain signals at peripheral terminals via alpha-2 adrenoceptors without producing the undesirable central side effects observed following the systemic administration. The ineffectiveness of topical ketamine to block topical clonidine antinociceptive tolerance suggests that peripheral NMDA receptors do not mediate local clonidine antinociceptive tolerance. PMID- 15363884 TI - The anticipation of pain modulates spatial attention: evidence for pain specificity in high-pain catastrophizers. AB - Recent studies have suggested that the anticipation of pain may modulate spatial attention. However, it is possible that this modulation reflects a general effect of anticipating somatosensory stimulation, without being pain-specific. In the present study, we therefore compared the effect of the anticipation of somatosensory stimulation on spatial attention between two groups, using conditioned signals in a spatial cueing paradigm. In the pain group, signals predicted painful electrocutaneous stimulation, whereas in the control group, signals predicted non-painful vibrotactile stimulation. Tests between both groups showed that attentional engagement was equally facilitated by the anticipation of somatosensory stimulation in both groups. Interestingly, disengagement of attention was more retarded by the anticipation of pain than by the anticipation of non-painful vibrotactile stimulation in participants high in catastrophic thinking about pain. Theoretical and clinical implications of these findings are discussed. PMID- 15363885 TI - Differential distribution of KChIPs mRNAs in adult mouse brain. AB - The K(+) channel interacting proteins (KChIPs1-4) interact with and modulate activity and trafficking of Kv4 potassium channels. We report here the distribution of KChIPs in adult mouse brain. KChIP1 was expressed in a subpopulation of neurons widely distributed in brain and enriched in Purkinje cells of the cerebellum and in the reticular thalamic and medial habenular nuclei. KChIP2 transcripts were highly expressed in layer IV of the cerebral cortex and in striatum and hippocampus, but expressed at low levels in cerebellum. KChIP3 transcripts were detected primarily in the layer V and deep layer VI of the cerebral cortex, the hippocampus, and the entire cerebellum. KChIP4 was highly expressed by neurons in layers II-IV of cortex and in hippocampus, thalamus and the Purkinje cell layer of the cerebellum. Collectively, different KChIPs appear to be expressed by selected neuronal populations in different brain regions where expression of Kv4.2 and Kv4.3 have been reported. These findings support the likelihood of functional interactions between KChIPs and Kv4 K(+) channels in brain. PMID- 15363886 TI - Identification of native rat cerebellar granule cell currents due to background K channel KCNK5 (TASK-2). AB - The TWIK-related, Acid Sensing K (TASK-2; KCNK5) potassium channel is a member of the tandem pore (2P) family of potassium channels and mediates an alkaline pH activated, acid pH-inhibited, outward-rectified potassium conductance. In previous work, we demonstrated TASK-2 protein expression in newborn rat cerebellar granule neurons (CGNs). In this study, we demonstrate TASK-2 functional expression in CGNs as a component of the pH-sensitive, volatile anesthetic-potentiated, standing-outward potassium conductance (I(K,SO)). Using excised, inside-out patch-clamp technique, we studied CGNs grown in primary culture. We identified four distinct, noninactivating single channel potassium conductances, Types 1-4. Types 1-3 have previously been attributed to TASK-1 (KCNK3), TASK-3 (KCNK9) and TASK-1/TASK-3 heteromers, and TREK-2 (KCNK10) 2P potassium channel function, respectively; however, the Type 4 conductance is currently unassigned. Previous studies demonstrated that Type 4 single channel activity is potentiated by extracellular, alkaline pH and cytoplasmic arachidonic acid (10-20 microM) and inhibited by cytoplasmic tetraethylammonium (TEA; 1 mM). We determined that heterologously expressed TASK-2 channels have single channel gating, conductance properties and pH sensitivity identical to the Type 4 conductance. Additionally, we found that TASK-2 single channel activity, like the Type 4 conductance is potentiated by cytoplasmic arachidonic acid (20 microM) and inhibited by cytoplasmic TEA (1 mM). We conclude that TASK-2 mediates the Type 4 single channel conductance in CGNs as a component of I(K,SO). PMID- 15363887 TI - Increased GAD67 mRNA levels are correlated with in vivo GABA synthesis in the MPTP-treated catecholamine-depleted goldfish brain. AB - The role of catecholamine neuronal input on GABAergic activity in the hypothalamus, telencephalon, optic tectum, and cerebellum was investigated in early recrudescent female goldfish (Carassius auratus). A new quantitative technique was developed and validated, permitting concomitant quantification and correlational analysis of glutamic acid decarboxylase 65 (GAD65), GAD67, and GAD3 mRNA levels and in vivo GABA synthesis. Catecholamine depletion was achieved by the administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; 50 microg/g body weight) and dopamine (DA) depletion verified by HPLC. Endogenous GABA levels were increased by intraperitoneal administration of gamma-vinyl GABA (GVG; 300 microg/g body weight), an inhibitor of the GABA catabolic enzyme GABA transaminase. Treatment with MPTP resulted in a greater than twofold increase in GABA synthesis rate in the optic tectum and telencephalon. The increase in GABA synthesis rate was highly correlated with an increase in GAD67, but not GAD65 or GAD3 mRNA levels. These results suggest that catecholaminergic input exerts inhibitory effects on GABA synthesis rates through the modulation of GAD67 in the optic tectum and telencephalon. Together with previously published observations in rodents and primates, it is suggested that catecholaminergic control of GABA synthesis must have evolved more than 200 million years ago, before the emergence of the teleost fishes. PMID- 15363888 TI - Mouse homolog of KIAA0143 protein: hearing deficit induces specific changes of expression in auditory brainstem neurons. AB - Hearing deficit induced by mechanical cochlear damage, intense noise or ototoxic drugs produces a variety of structural and functional changes in the inner ear and the auditory brainstem. In the present study, we identified a novel gene that has activity dependent plasticity in the superior olivary complex by using suppression subtractive hybridization. We cloned a gene that encodes mouse homolog of KIAA0143 protein, one derived from a series of unidentified human genes. This gene termed mKIAA0143 shows differential expression of mRNA in the lateral superior olive between mice with hearing deficit and those with normal hearing ability. The mRNA thus obtained encodes a unique membrane-bound protein that consists of 819 amino acids. The gene locus was mapped using genomic DNA databases to the mouse chromosome 15D1. Green fluorescent protein-tagged mKIAA0143 was expressed in COS-1 cells. It was amply seen in the cellular plasma membrane. PMID- 15363889 TI - LE-PAS, a novel Arnt-dependent HLH-PAS protein, is expressed in limbic tissues and transactivates the CNS midline enhancer element. AB - A growing family of helix-loop-helix PAS (HLH-PAS) transcription factors has emerged recently. These proteins form heterodimers and are involved in adaptation to environmental or physiologic stresses as well as in embryonic development. We describe the cloning and characterization of a novel HLH-PAS protein termed LE PAS (limbic-enriched PAS) in mouse. LE-PAS consists of 802 amino acids and has a predicted molecular mass of 87.4 kDa. In adult mice, it is expressed exclusively in brain. Northern blot and in situ hybridization analysis indicate that LE-PAS is expressed in the limbic system and olfactory bulb. By in vitro overexpression in COS-7, cells we found that LE-PAS is a nuclear protein. Reporter gene analysis indicated that LE-PAS transactivates the CNS midline enhancer (CME) motif but not the xenobiotic response element in an Arnt-dependent fashion and without prior activation of LE-PAS protein. Our observation suggests that LE-PAS shares a similar mode of function with HLH-PAS proteins such as single minded or trachealess indicating that LE-PAS also has constitutive or developmental functions which may be critical for regulating the transcriptional control of limbic patterning and function. PMID- 15363890 TI - Increases in melanin-concentrating hormone and MCH receptor levels in the hypothalamus of dietary-obese rats. AB - Melanin-concentrating hormone (MCH) is a hypothalamic neuropeptide that stimulates feeding and increases body weight in rodents. We studied the role of the system in energy homeostasis and its regulation by the satiety signals, leptin and insulin. We used real-time PCR to measure the hypothalamic expression of MCH and its receptor (MCHR1) in two contrasting models of altered nutritional status, namely, obesity induced by 8 weeks' voluntary overeating and food restriction for 10 days. Diet-fed rats were stratified according to final total fat-pad mass into a 'high fat gain' group (HG) and 'low fat gain' group (LG). MCH mRNA levels were increased by 31% (p>0.05) and 49% (p<0.05) in the LG and HG, respectively, compared with controls. MCHR1 mRNA levels rose by 118% in the LG (p<0.01) and 85% in the HG (p<0.01). There were significant positive correlations (p<0.05) between plasma leptin concentration and both MCH and MCHR1 mRNA levels, and between plasma insulin and MCHR1 expression. A positive correlation was also observed between MCH and MCHR1 mRNA levels (p<0.05). Food-restricted rats showed no significant alterations in the levels of either MCH mRNA or MCHR1 mRNA. In a second experiment, we measured MCH peptide levels in five discrete hypothalamic areas of dietary-obese rats. MCH concentrations were significantly increased in the arcuate nuclei of the HG (p<0.05) and the paraventricular nuclei of both the LG (p<0.05) and HG (p<0.05), compared with their lean counterparts. These results suggest that the MCH system becomes more active in dietary obesity and could be involved in enhancing appetite for palatable food. The possibility that MCH and MCHR1 expression are positively regulated by leptin and insulin, which normally inhibit feeding, is a putative explanation for how appetite for palatable food is able to override mechanisms that prevent the development of obesity. PMID- 15363891 TI - Glutamate mediates cell death and increases the Bax to Bcl-2 ratio in a differentiated neuronal cell line. AB - Excessive stimulation of the NMDA receptor by glutamate induces cell death and has been implicated in the development of several neurodegenerative diseases. While apoptosis plays a role in glutamate-mediated toxicity, the mechanisms underlying this process have yet to be completely determined. Recent evidence has shown that exposure to excitatory amino acids regulates the expression of the antiapoptotic protein, Bcl-2, and the proapoptotic protein, Bax, in neurons. Since it has been suggested that the ratio of Bax to Bcl-2 is an important determinant of neuronal survival, the reciprocal regulation of these Bcl-2 family proteins may play a role in the neurotoxicity mediated by glutamate. Here, we have used a differentiable neuronal cell line, N1E-115, to investigate the molecular properties of glutamate-induced cell death. Annexin V staining was used to determine apoptotic cell death between 0 and 5 days differentiation with DMSO/low serum. Immunoblot analysis was used to determine whether the expression of Bcl-2 or Bax was modulated during the differentiation process. Bcl-2 protein levels were increased during maturation while Bax expression remained unchanged. Maximum Bcl-2 expression was observed following 5 days of differentiation. Examination of Bcl-2 and Bax following glutamate treatment revealed that the expression of these proteins was inversely regulated. Exposure to glutamate (0.001-10 mM) for 20+/-2 h resulted in a dose-dependent decrease in cell survival (as measured by MTT analysis) that was maximal at 10 mM. These results further support the role of apoptosis in glutamate-mediated cell death. Furthermore, a significant decrease in Bcl-2 levels was observed at 1 mM and 10 mM glutamate (32.1%+/-4.8 and 33.7+/-12.8%, respectively) while a significant upregulation of Bax expression (88.2+/-17.9%) was observed at 10 mM glutamate. Interestingly, Bcl 2 and Bax levels in cells treated with glutamate from 12-24 h were not significantly different from those of control. Taken together, these findings provide additional evidence for the reciprocal regulation of Bcl-2 and Bax expression by glutamate and suggest that neuronal excitotoxicity may, in part, result from the inverse regulation of these proteins. PMID- 15363892 TI - Functional analysis of glutamate transporters in excitatory synaptic transmission of GLAST1 and GLAST1/EAAC1 deficient mice. AB - The high affinity, Na(+)-dependent, electrogenic glial L-glutamate transporters GLAST1 and GLT1, and two neuronal EAAC1 and EAAT4, regulate the neurotransmitter concentration in excitatory synapses of the central nervous system. We dissected the function of the individual transporters in the monogenic null allelic mouse lines, glast1(-/-) and eaac1(-/-), and the derived double mutant glast(-/-)eaac1( /-). Unexpectedly, the biochemical analysis and the behavioral phenotypes of these null allelic mouse lines were inconspicuous. Inhibition studies of the Na(+)-dependent glutamate transport by plasma membrane vesicles and by isolated astrocytes of wt and glast1(-/-) mouse brains indicated the pivotal compensatory role of GLT1 in the absence particularly of GLAST1 and GLAST1 and EAAC1 mutant mice. In electrophysiological studies, the decay rate of excitatory postsynaptic currents (EPSCs) of Purkinje cells (PC) after selective activation of parallel and climbing fibers proved to be similar in wt and eaac1(-/-), but was significantly prolonged in glast1(-/-) PCs. Bath application of the glutamate uptake blocker SYM2081 prolonged EPSC decay profiles in both wt and double mutant glast1(-/-)eaac1(-/-) PCs by 286% and 229%, respectively, indicating a prominent role of compensatory glutamate transport in shaping glast1(-/-)eaac1(-/-) EPSCs. PMID- 15363893 TI - Endophilin A3 forms filamentous structures that colocalise with microtubules but not with actin filaments. AB - Endophilin A3 is a member of the endophilin family of proteins, thought to play a role in the formation of clathrin-coated vesicles from the plasma membrane in the process of clathrin-mediated endocytosis. We investigated the localisation of both endogenous and overexpressed endophilin A3 within mammalian cells. Endophilin A3 demonstrated a complex cellular distribution with bright punctate structures and filamentous strands superimposed on a diffuse cytoplasmic background. The endophilin A3 structures did not colocalise with mitochondria, endoplasmic reticulum or lysosomes. Direct immunolocalisation and cytoskeletal perturbation studies showed that the filamentous structures were more likely to be colocalised with microtubules than actin filaments. We therefore propose that endophilin A3 has a role in transport along or as part of the structure of microtubules, in addition to its suggested role in endocytosis. PMID- 15363894 TI - Proteomic identification of brainstem cytosolic proteins in a neuropathic pain model. AB - Neuropathic pain involves co-regulation of many genes and their translational products in both peripheral and central nervous system. We used proteomics approaches to investigate expressional changes in cytosolic protein levels in rat brainstem tissues following ligation of lumbar 5 and 6 (L5, L6) spinal nerves, which generates a model of peripheral neuropathic pain (NP). Proteins from brainstem tissue homogenates of NP and SHAM animals were fractionated by two dimensional (2-DE) gel electrophoresis to produce a high-resolution map of the brainstem soluble proteins. Proteins showing altered expression levels between NP and SHAM were selected. Isolated proteins were in-gel trypsin-digested and the resulting peptides were analyzed by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry. Using the mass spectrometric data, we were able to identify 17 proteins of interest through searches of the Swiss Prot and NCBi nonredundant protein sequence database. Several of the identified proteins, including fatty acid binding protein-brain (FABP-B), major histocompatibility complex (MHC) class 1, T-cell receptor (TCR) alpha chain, and interleukin-1 (IL-1), showed significantly higher levels in the NP rat brainstem. Proteomic analysis has identified several proteins with differential expression levels in NP as compared to SHAM. However, the function of the proteins identified is postulated; therefore, further experiments are required to determine the true role of each protein in NP. PMID- 15363895 TI - The endocytotic pathway is required for increased A beta 42 secretion during apoptosis. AB - Secretion and progressive cerebral accumulation of beta-amyloid peptides (A beta), which derive by endoproteolytic ('amyloidogenic') processing of beta amyloid precursor protein (APP), are felt to represent collectively an early and necessary event in the pathogenesis of Alzheimer's disease. APP amyloidogenic processing can occur via secretory or endocytotic pathways, but the relative contribution of these pathways to A beta secretion remains to be established. The effect of apoptosis on amyloidogenic processing and A beta secretion similarly is incompletely understood. We tested the hypothesis that APP processing by the endocytotic pathway represents a stress-related neural cell response, by comparing A beta secretion after induction of apoptosis in PC12 cells transfected either for endocytosis-competent or -deficient APP. Newly prepared adenoviral vectors encompassing targeted mutagenesis of the cytoplasmic tail YENP tetrapeptide sequence, which serves as the principal APP internalization signal, were used to express endocytosis-deficient holoprotein. We report that the endocytotic pathway is required for the generation and secretion of A beta 42, and that secretion of this neurotoxic peptide increases significantly during apoptosis. We demonstrate additionally that more A beta 40 apparently is generated in secretory compartments during apoptosis when APP processing by the endocytotic pathway is impaired. PMID- 15363896 TI - Genome duplication led to highly selective expansion of the Arabidopsis thaliana proteome. AB - Multiple ancient genome duplications in Arabidopsis thaliana provide unique opportunities to assess factors that influence the fates of duplicated genes. We have found that genes retained in duplicate following one round of genome duplication are significantly more likely to be retained in duplicate again after a subsequent genome duplication. Genes retained in duplicate form a functionally biased set and include a significant over-representation of genes involved in the regulation of transcription. PMID- 15363897 TI - Retrotransposition of Alu elements: how many sources? AB - It is generally thought that only a few Alu elements are capable of retrotransposition and that these 'master' sources produce inactive copies. Here, we use a network phylogenetic approach to demonstrate that recently integrated human-specific Alu subfamilies typically contain 10-20% of secondary source elements that contributed 20-40% of all subfamily members. This multiplicity of source elements provides new insight into the remarkably successful amplification strategy of the Alu family. PMID- 15363898 TI - Protein domains enriched in mammalian tissue-specific or widely expressed genes. AB - In multicellular organisms some genes are expressed in essentially all tissues, whereas others are expressed predominantly in only one or a few tissues. In this study, we investigate the relationship between the tissue-specificity of gene expression and the type of protein encoded. We find that many protein domains are found to be enriched in either tissue-specific or widely expressed genes. Domains enriched in tissue-specific genes tend to be metazoan-specific; these same domains are also enriched in genes that are not essential for cell viability. These findings identify families of proteins that are probably used in the development or terminal differentiation of many different tissue types. PMID- 15363899 TI - Negative selection pressure against premature protein truncation is reduced by alternative splicing and diploidy. AB - The importance of alternative splicing in many genomes has raised interesting questions about its role in evolution. We analyzed 13 384 full-length transcript isoforms from human and 2227 isoforms from mouse to identify sequences containing premature termination codons (PTCs) that are likely targets of mRNA nonsense mediated decay. We found that alternatively spliced isoforms have a much higher frequency of PTCs (11.1%) compared with the major transcript form of each gene (3.7%). On the X chromosome, which is generally expressed as a single copy, the overall PTC rate was much lower (3.5%, versus 8.9% on diploid autosomes), and the effect of alternative splicing was enhanced. Thus, diploidy and alternative splicing each increased tolerance for PTC by about threefold, as approximately additive effects. These data suggest that nonsense mediated decay might itself reduce negative selection pressure during evolution, via rapid degradation of aberrant transcripts that might yield dominant negative phenotypes. PMID- 15363900 TI - Conserved regulatory motifs in bacteria: riboswitches and beyond. AB - We present a computational approach that identifies regulatory elements conserved across phylogenetically distant organisms. Intergenic regulatory regions were clustered by orthology of the adjacent genes, and an iterative process was applied to search for significant motifs, enabling new elements of the putative regulon to be added in each cycle. With this approach, we identified highly conserved riboswitches and the Gram positive T-box. Interestingly, we identified many other regulatory systems that appear to depend on conserved RNA structures. PMID- 15363901 TI - Is avian sex determination unique?: clues from a warbler and from chickens. PMID- 15363902 TI - Subfunction partitioning, the teleost radiation and the annotation of the human genome. AB - Half of all vertebrate species are teleost fish. What accounts for this explosion of biodiversity? Recent evidence and advances in evolutionary theory suggest that genomic features could have played a significant role in the teleost radiation. This review examines evidence for an ancient whole-genome duplication (tetraploidization) event that probably occurred just before the teleost radiation. The partitioning of ancestral subfunctions between gene copies arising from this duplication could have contributed to the genetic isolation of populations, to lineage-specific diversification of developmental programs, and ultimately to phenotypic variation among teleost fish. Beyond its importance for understanding mechanisms that generate biodiversity, the partitioning of subfunctions between teleost co-orthologs of human genes can facilitate the identification of tissue-specific conserved noncoding regions and can simplify the analysis of ancestral gene functions obscured by pleiotropy or haploinsufficiency. Applying these principles on a genomic scale can accelerate the functional annotation of the human genome and understanding of the roles of human genes in health and disease. PMID- 15363903 TI - Bringing the role of mRNA decay in the control of gene expression into focus. AB - The process of mRNA decay is integral to the post-transcriptional control of gene expression. The enzymes of the pathway have been identified, and now several laboratories have found that the mRNA decay machinery is localized to discrete cytoplasmic foci whose existence had not been suspected previously. In addition, we can now see that mRNA turnover is a means to coordinate gene expression, first through integration with control of transcription, export and translation of mRNAs, and second through enabling mRNAs involved in similar processes to decay at similar rates. These and other aspects of the field are discussed. PMID- 15363904 TI - Patterns in evolution: veins of the Drosophila wing. AB - The development of the Drosophila wing is a classical model for studying the genetic control of tissue size, shape and patterning. A detailed picture of how positional information is interpreted by cells in the imaginal disc and translated into the adult wing vein pattern has recently emerged. It highlights the central role of dose-dependent activation of distinct cell transcription programs in response to the Hedgehog (Hh) and Decapentaplegic (Dpp) morphogens, as well as an early role of Notch signalling, in connecting the positioning of vein primordia and vein differentiation proper. The biochemical basis of the cross-talk that operates between these different signalling pathways is less well understood. New strategies made possible by the genome sequencing of several insect models should provide an important complement to the knowledge obtained from >60 years of genetic studies. PMID- 15363905 TI - Pathogenic RNA repeats: an expanding role in genetic disease. AB - Fragile X mental retardation and Friedreich's ataxia were among the first pathogenic trinucleotide repeat disorders to be described in which noncoding repeat expansions interfere with gene expression and cause a loss of protein production. Invoking a similar loss-of-function hypothesis for the CTG expansion causing myotonic dystrophy type 1 (DM1) located in the 3' noncoding portion of a kinase gene was more difficult because DM is a dominantly inherited multisystemic disorder in which the second copy of the gene is unaffected. However, the discovery that a transcribed but untranslated CCTG expansion causes myotonic dystrophy type 2 (DM2), along with other discoveries on DM1 and DM2 pathogenesis, indicate that the CTG and CCTG expansions are pathogenic at the RNA level. This review will detail recent developments on the molecular mechanisms of RNA pathogenesis in DM, and the growing number of expansion disorders that might involve similar pathogenic RNA mechanisms. PMID- 15363906 TI - Coupling the mitochondrial transcription machinery to human disease. AB - Mitochondria are the central processing units for cellular energy metabolism and, in addition to carrying out oxidative phosphorylation, regulate important processes such as apoptosis and calcium homeostasis. Because mitochondria possess a genome that is central to their multiple functions, an understanding of the mechanism of mitochondrial gene expression is required to decipher the many ways mitochondrial dysfunction contributes to human disease. Towards this end, two human transcription factors that are related to rRNA methyltransferases have recently been characterized, providing new insight into the mechanism of mitochondrial transcription and a novel link to maternally inherited deafness. Furthermore, studies in the Saccharomyces cerevisiae model system have revealed a functional coupling of transcription and translation at the inner mitochondrial membrane, where assembly of the oxidative phosphorylation system commences. Defects in an analogous coupling mechanism in humans might underlie the cytochrome oxidase deficiency that causes a form of Leigh Syndrome. PMID- 15363907 TI - Maximizing aesthetics in lateral-tension abdominoplasty and body lifts. AB - The high-lateral-tension abdominoplasty addresses the practical and theoretic faults of standard abdominoplasty design. Key features include limited direct undermining, increased lateral skin resection with highest-tension wound closure along lateral limbs, two-layer superficial fascial system repair, and significant truncal liposuction when needed. The high-lateral-tension design limits the unfavorable features of standard abdominoplasty and produces balanced natural aesthetic contours. The high-lateral-tension abdominoplasty is the foundation for treatment of more generalized relaxation problems in the circumferential trunk and thighs. For more significant thigh laxity and buttock ptosis, the lateral tension abdominoplasty is combined with the transverse thigh/buttock lift to produce the lower body lift #2. PMID- 15363908 TI - The overlap of lipoplasty and abdominoplasty: indication, classification, and treatment. AB - The aesthetic and functional treatment of the abdominal region has always been one of the main concerns of plastic surgeons. Some patients belong in a "gray area" where there is no clear indication of which technique should be applied. Individual variations in anatomy should be the first consideration in determining the correct procedure. The Type I abdomen can be treated by lipoplasty alone. Types II, III, and IV are considered to be in the "gray area," because we can use different approaches and techniques based not only on this classification system but also on patients' preferences. Types V and VI can be treated by dermolipectomy with or without lipoplasty. PMID- 15363909 TI - The male abdominoplasty. AB - Men have a larger body surface area, distinguishing physical characteristics, and unique aesthetic concerns that present a different therapeutic challenge from women. The primary area of disappointment in males is the inability to alter the intra-abdominal submuscular fat compartment where fat redistributes with advancing age, resulting in enlarged abdominal girth. In general, men who have not experienced large weight fluctuations present with skin of good quality and tone and can benefit from liposuction surgery; those who have poor skin tone, with or without rectus muscle diastasis, may be candidates for a full abdominoplasty or a pannilectomy with liposuction. These procedures represent the overwhelming majority of abdominal contour operations performed in males. PMID- 15363910 TI - "Marriage" abdominoplasty: body contouring with limited scars combining mini abdominoplasty and liposuction. AB - The "marriage" or union of aggressive truncal liposuction with modified abdominoplasty techniques has been applied by the authors to treat patients with abdominal deformities marked by lower abdominal skin excess, abdominal muscle laxity, and excess adipose tissue on the abdominal wall and in adjacent contours. In the appropriately selected patient this form of "mini-abdominoplasty" has resulted in excellent contour improvement and a more rapid return to life activities than is seen with "full" abdominoplasty. In the authors' experience, it is applicable to the majority of patients (over 50%) presenting for abdominal contour improvement. PMID- 15363911 TI - Progressive tension sutures in abdominoplasty. AB - The authors describe a simple, safe, and effective way to reduce complications related to abdominoplasty. By using progressive tension sutures, a suture technique to advance and anchor the abdominal flap, they have routinely combined abdominal liposuction and abdominoplasty, with no resulting seroma formation or distal flap necrosis. No drains are used in this technique, and early ambulation in an upright posture is encouraged. PMID- 15363912 TI - The state of the art in bariatric surgery for weight loss in the morbidly obese patient. AB - Bariatric surgery is a safe and effective method for achieving durable weight loss for patients with morbid obesity. Gastric restrictive procedures include vertical banded gastroplasty and gastric banding. Malabsorptive procedures include long-limb gastric bypass, biliopancreatic diversion, and biliopancreatic diversion with duodenal switch. The gastric bypass has features of both restriction and malabsorption. The laparoscopic approach to bariatric surgery has substantially improved postoperative recovery. Careful patient selection and preoperative work-up are extremely important. A number of medical comorbidities are improved after surgically-induced weight loss. PMID- 15363913 TI - Perioperative management of the post-gastric-bypass patient presenting for body contour surgery. AB - Bariatric surgery has evolved as an effective and relatively safe treatment for morbid obesity. With nearly every region of the body as a potential operative site and an unprecedented number of surgical procedures available, we must give attention to thoughtful perioperative management. Bariatric surgery is a life changing event for the morbidly obese patient, and the body contouring that follows weight loss often has an equally profound effect. Plastic surgeons must strive to maintain the highest level of safety in this pursuit. The authors address issues surrounding preoperative evaluation and measures to minimize the risk of complications. PMID- 15363914 TI - Truncal body contouring surgery in the massive weight loss patient. AB - Body contouring of massive weight loss patients is the latest frontier in plastic surgery. It is important to become familiar with the presentation of these patients and with the delineation and effective treatment of their deformities. Because they usually present with circumferential excess, circumferential belt lipectomy treats their truncal contour as a unit. This article details the preoperative work-up, intraoperative technique, and postoperative course for the belt-lipectomy patient. Results and how to predict them are discussed. Complications are enumerated and suggestions offered for avoiding them. PMID- 15363916 TI - Pelvic fracture patterns and their corresponding angiographic sources of hemorrhage. AB - The association between pelvic arterial injuries and associated fracture patterns has been investigated directly in a post mortem study and indirectly in an earlier clinical study evaluating an overall management approach to pelvic fractures. One previous report has correlated the angiographic findings of a group of patients with pelvic ring disruptions. The authors' study was designed to further define the anatomic sites of hemorrhage associated with specific pelvic ring injury patterns and evaluate patient survival in each of these groups. PMID- 15363917 TI - The role of pelvic angiography in evaluation and management of pelvic trauma. AB - Controversy exists about the indications and optimal timing of angiography in hemodynamically unstable patients with severe pelvic fractures. Recommendations from published studies are limited by small numbers of patients. In this article the recommended indications, timing, and drawbacks to angiography are reviewed. PMID- 15363918 TI - Instability of the posterior pelvic ring associated with disruption of the pubic symphysis. AB - The patient with instability of the posterior pelvic ring and disruption of the pubic symphysis is a challenging problem for orthopedic surgeons. These represent high-energy injuries and management of these patients requires a multidisciplinary team approach. The orthopedic surgeon's understanding of the injury patterns, their associated injuries, and effective treatment strategies can reduce morbidity and mortality for patients. PMID- 15363919 TI - Soft tissue injuries associated with pelvic fractures. AB - Significant soft tissue injuries often occur as part of high-energy injuries to the pelvis. These soft tissue injuries must be recognized and considered when implementing a treatment plan if complications are to be minimized. Vigilance in diagnosing open fractures must be maintained. Patients with these injuries must be managed aggressively, because they are at high risk for complications and death. Closed pelvic and acetabular fractures also may include soft tissue injury that requires special consideration. Careful evaluation and management of the soft tissues aids in determining appropriate techniques for reduction and fixation of the associated fractures. PMID- 15363920 TI - Trochanteric osteotomy for acetabular fractures and proximal femur fractures. AB - Anatomic reconstruction of the articular surface is a primary goal of internal fixation of fractures of the hip joint. The quality of the reduction correlates with long-term outcomes. The traditional Kocher-Langenbeck and the ilioinguinal approaches, however, rely on extra-articular assessment of the quality of the reduction. Ganz et al described a technique of trochanteric osteotomy combined with a Kocher-Langenbeck approach that allows direct visualization of the joint without the risk for avascular necrosis of the femoral head. This article reviews the indications of this approach in the treatment of fractures around the hip joint. PMID- 15363921 TI - Femoral head fractures: diagnosis, management, and complications. AB - Femoral head fractures occur with a reported incidence of 6%-16% after hip dislocation. Even the largest series have evaluated only a few dozen patients, often collecting experience from multiple centers. This article provides the clinician with current information to guide the diagnosis and treatment of this uncommon condition. PMID- 15363922 TI - Subtrochanteric femur fractures. AB - Subtrochanteric femur fractures have demanded special consideration in orthopaedic traumatology, given the high rate of complications associated with their management. The intense concentration of compression, tensile, and torsional stresses and decreased vascularity of the region has challenged orthopaedists with problems of malunion, delayed union, and nonunion resulting from loss of fixation, implant failure, and iatrogenic devascularization of the operative exposure. Only recently has a better understanding of fracture biology, reduction techniques, and biomechanically improved implants allowed for subtrochanteric fractures to be addressed with consistent success. PMID- 15363923 TI - A prospective, modernized treatment protocol for periprosthetic femur fractures. AB - This article reports a prospective series of periprosthetic femur fractures in 33 patients treated with a modernized fracture treatment protocol. Some form of operative treatment was selected prospectively based on the categorization of the fracture by the Duncan-Vancouver and Beals-Tower Oregon classification systems. Fractures in which the prosthesis-bone interface was stable were treated with open reduction and internal fixation of the fracture around the stable implant. Unstable prosthesis-bone interfaces required removal of the primary prosthesis and revision to an uncemented long-stem prosthesis after stabilization of the femoral fracture with plates or allograft struts. Complications were minimal over an average follow-up period of 28.3 months; restoration of function was predictable. PMID- 15363924 TI - Injuries to the hip joint in frontal motor-vehicle crashes: biomechanical and real-world perspectives. AB - Hip fractures and dislocations in frontal crashes are of substantial concern to clinicians and automotive safety engineers because of the frequency at which hip injuries occur and the associated potential for long-term disability. Impacts to the flexed knees of unembalmed cadavers under loading conditions similar to those that occur in frontal crashes of newer-model vehicles indicate that the hip is the weakest part of the knee-thigh-hip complex and that hip injury tolerance is reduced by hip flexion and adduction from a typical driving posture. These results are being used to develop new knee-thigh-hip injury assessment criteria for use in anthropomorphic test devices (crash test dummies). PMID- 15363926 TI - Simulation study on effects of signaling network structure on the developmental increase in complexity. AB - The developmental increase in structural complexity in multicellular lifeforms depends on local, often non-periodic differences in gene expression. These, in turn, depend on a network of gene-gene interactions coded within the organismal genome. To see what architectural features of a network (size, connectivity, etc.) affect the likelihood of patterns with multiple cell types (i.e. patterns where cells express > or = 3 different combinations of genes), developmental pattern formation was simulated in virtual blastoderm embryos with small artificial genomes. Several basic properties of these genomic signaling networks, such as the number of genes, the distributions of positive (inductive) and negative (repressive) interactions, and the strengths of gene-gene interactions were tested. The results show that the frequencies of complex and/or stable patterns depended not only on the existence of negative interactions, but also on the distribution of regulatory interactions: for example, coregulation of signals and their intracellular effectors increased the likelihood of pattern formation compared to differential regulation of signaling pathway components. Interestingly, neither quantitative differences in strengths of signaling interactions nor multiple response thresholds to different levels of signal concentration (as in morphogen gradients) were essential for formation of multiple, spatially unique "cell types". However, those combinations of architectural features that greatly increased the likelihood for pattern complexity tended to decrease the likelihoods for pattern stability and developmental robustness. Nevertheless, elements of complex patterns (e.g. genes, cell type order within the pattern) could differ in their developmental robustness, which may be important for the evolution of complexity. The results show that depending on the network structure, the same set of genes can produce patterns of different complexity, robustness and stability. Because of this, the evolution of metazoan complexity with a combinatorial code of gene regulation may have depended at least as much on selection for favorable distribution of connections between existing developmental regulatory genes as on the simple increase in numbers of regulatory genes. PMID- 15363927 TI - A two variable delay model for the circadian rhythm of Neurospora crassa. AB - A two variable model with delay in both the variables, is proposed for the circadian oscillations of protein concentrations in the fungal species Neurospora crassa. The dynamical variables chosen are the concentrations of FRQ and WC-1 proteins. Our model is a two variable simplification of the detailed model of Smolen et al. (J. Neurosci. 21 (2001) 6644) modeling circadian oscillations with interlocking positive and negative feedback loops, containing 23 variables. In our model, as in the case of Smolen's model, a sustained limit cycle oscillation takes place in both FRQ and WC-1 protein in continuous darkness, and WC-1 is anti phase to FRQ protein, as observed in experiments. The model accounts for various characteristic features of circadian rhythms such as entrainment to light dark cycles, phase response curves and robustness to parameter variation and molecular fluctuations. Simulations are carried out to study the effect of periodic forcing of circadian oscillations by light-dark cycles. The periodic forcing resulted in a rich bifurcation diagram that includes quasiperiodicity and chaotic oscillations, depending on the magnitude of the periodic changes in the light controlled parameter. When positive feedback is eliminated, our model reduces to the generic one dimensional delay model of Lema et al. (J. Theor. Biol. 204 (2000) 565), delay model of the circadian pace maker with FRQ protein as the dynamical variable which represses its own production. This one-dimensional model also exhibits all characteristic features of circadian oscillations and gives rise to circadian oscillations which are reasonably robust to parameter variations and molecular noise. PMID- 15363928 TI - Control of the pathogenic conformational change of the prion protein by an attractor of low order. AB - The molecular vibrations of the infectious part of the prion protein has been studied by the methods of nonlinear dynamics using NMR-data of the protein in solution to test a new hypothesis for the nature of the pathogenic isomerization. The result was that the conformational change accompanying the conversion of the physiological form (PrP(c)) to the pathological (PrP(Sc)) one displays the characteristic properties of an attractor of the dimension 2.7+/-0.2, whereas the dimensions of the C-terminal, potentially infectious half of the physiological and the pathological forms are 5.3+/-0.3 and 3.9+/-0.3, resp. A plot of the average RMS of the vibrations per atom of the amino acids along the peptide chain suggests a pivotal role of E(167) and D(196). The vibrations of these residues are strongly dampened by the pathogenic conformational change suggesting that the neutralization to the hydrophobic form facilitates the isomerisation. This observation lends credence to the hypothesis that the pathological conformational change is released by the charge neutralization of the abundant, basic side chains lining the cleft in PrP(c) by a pK-shift caused by a transient ion flux from an action potential or by an RNA ligand. The attractor ensures that only the two conformers PrP(c) and PrP(Sc)-monomer, which are switching the polyadenylation of mRNA for synaptic proteins on and off, prevail. Some xenogenic PrP(Sc) seem to have a tendency to polymerize irreversibly, which is likely to inhibit the translation, thus killing neurons. The autocorrelation function is dampened (long correlation length for the 3 cases), the Poincare plot seems to show the cross-section of a dense attractor for PrP(Sc) and a loose one for PrP(c), the Lyapunov exponent is positive and the power spectrum is broad. The Hurst plots of PrP(Sc) and PcP(c) show monofractality. The attractor hypothesis offers a supplement, or an alternative, to the current, inconclusive ideas of the nature of the PrP isomerization. The identification of the control factors may permit the reversal of the fatal conformational change. The dynamic parameters were checked by the interpoint distance method of Judd and by an analysis of the Fourier-transformed data. PMID- 15363929 TI - A mechanistic model for eukaryotic gradient sensing: spontaneous and induced phosphoinositide polarization. AB - The crawling movement of cells in response to a chemoattractant gradient is a complex process requiring coordination of various subcellular activities. Although a complete description of the mechanisms underlying cell movement remains elusive, the very first step of gradient sensing, enabling the cell to perceive the imposed gradient, is becoming more transparent. The increased understanding of this step has been driven by the discovery that within 5-10 s of applying a weak chemoattractant gradient, membrane phosphoinositides such as PIP(3) localize at the front end of the cell. It is currently believed that the gradient sensing mechanism is precisely the mechanism leading to this localization. We have formulated a reaction-diffusion model based on the phosphoinositide cycle which predicts various responses of motile cells in addition to the phosphoinositide polarization induced by chemoattractant gradients. The responses include: (a) Polarized sensitivity wherein a polarized cell responds to a change in the direction of the gradient by turning its existing front. (b) Spontaneous polarization wherein cells polarize in a random direction even if the surrounding chemoattractant concentration is uniform. (c) Unique localization which refers to the formation of a unique polarity even in the face of multiple chemoattractant sources. The above responses preclude the hypothesis that the cell merely amplifies the external signal. Our model indicates that the cell must be viewed as a system that nonlinearly processes chemoattractant inputs. We show in particular that these seemingly complex dynamics can be explained very simply in terms of the instabilities and wavefront dynamics that are characteristic of the activator-inhibitor class of models. PMID- 15363930 TI - Mathematical modeling of granulocyte reconstitution after high-dose chemotherapy with stem cell support: effect of post-transplant G-CSF treatment. AB - Cancer patients treated with high-dose chemotherapy and autotransplanted with peripheral blood progenitor cells most often reconstitute neutrophils (> 0.5 x 10(9)c/l) 8-16 days after the initiation of treatment. By means of a mathematical model of human granulopoiesis, the present work assesses the effect of administering granulocyte colony stimulating factor (G-CSF) post-transplant to reduce engraftment time, and also assesses the effect of delaying initiation of G CSF treatment relative to a general schedule. Hematopoietic progenitor cells from 21 breast cancer patients were mobilized by chemotherapy followed by G-CSF injections. Purified CD34+ cells taken from the mobilized blood were infused 3 days after termination of chemotherapy. Patients were given subcutaneous injections of G-CSF post-transplant (5 microg/kg every 12 h). Neutrophil counts calculated from a mathematical model were compared with data from individual patients. These results were also compared with data and modeling results from a group of 19 lymphoma patients given no post-transplant G-CSF therapy. The observed engraftment times were associated with the number of CFU-GM cells in the reinfused blood graft and the administration of post-transplant G-CSF. The latter finding was most predominant in patients given < 5.0 x 10(5) CFU-GM/kg bw. These tendencies were well captured by the model. Interestingly, the model showed no major differences in time to engraft neutrophils if the initiation of G-CSF was postponed for up to 5 days after transplantation. Our findings indicate that the present mathematical model of neutrophil recovery following high-dose therapy correlates with clinical observations and can potentially be used to predict time to neutrophil recovery. PMID- 15363931 TI - Cooperativity of the oxidization of cysteines in globular proteins. AB - Based on the 639 non-homologous proteins with 2910 cysteine-containing segments of well-resolved three-dimensional structures, a novel approach has been proposed to predict the disulfide-bonding state of cysteines in proteins by constructing a two-stage classifier combining a first global linear discriminator based on their amino acid composition and a second local support vector machine classifier. The overall prediction accuracy of this hybrid classifier for the disulfide-bonding state of cysteines in proteins has scored 84.1% and 80.1%, when measured on cysteine and protein basis using the rigorous jack-knife procedure, respectively. It shows that whether cysteines should form disulfide bonds depends not only on the global structural features of proteins but also on the local sequence environment of proteins. The result demonstrates the applicability of this novel method and provides comparable prediction performance compared with existing methods for the prediction of the oxidation states of cysteines in proteins. PMID- 15363932 TI - Effects of neighbourhood size and connectivity on the spatial Continuous Prisoner's Dilemma. AB - The Prisoner's Dilemma, a two-person game in which the players can either cooperate or defect, is a common paradigm for studying the evolution of cooperation. In real situations cooperation is almost never all or nothing. This observation is the motivation for the Continuous Prisoner's Dilemma, in which individuals exhibit variable degrees of cooperation. It is known that in the presence of spatial structure, when individuals "play against" (i.e. interact with) their neighbours, and "compare to" ("learn from") them, cooperative investments can evolve to considerable levels. Here, we examine the effect of increasing the neighbourhood size: we find that the mean-field limit of no cooperation is reached for a critical neighbourhood size of about five neighbours on each side in a Moore neighbourhood, which does not depend on the size of the spatial lattice. We also find the related result that in a network of players, the critical average degree (number of neighbours) of nodes for which defection is the final state does not depend on network size, but only on the network topology. This critical average degree is considerably (about 10 times) higher for clustered (social) networks, than for distributed random networks. This result strengthens the argument that clustering is the mechanism which makes the development and maintenance of the cooperation possible. In the lattice topology, it is observed that when the neighbourhood sizes for "interacting" and "learning" differ by more than 0.5, cooperation is not sustainable, even for neighbourhood sizes that are below the mean-field limit of defection. We also study the evolution of neighbourhood sizes, as well as investment level. Here, we observe that the series of the interaction and learning neighbourhoods converge, and a final cooperative state with considerable levels of average investment is achieved. PMID- 15363933 TI - How should we define goodness?--reputation dynamics in indirect reciprocity. AB - Theory of indirect reciprocity is important in explaining cooperation between humans. Since a partner of a social interaction often changes, an individual should assess his partner by using social information such as reputation and make decisions whether to help him or not. To those who have 'good' social reputation does a player give aid as reciprocation, whereas he has to refuse to help those who have 'bad' reputation. Otherwise benefits of altruism is easily exploited by them. Little has been known, however, about the definition of 'goodness' in reputation. What kind of actions are and should be regarded as good and what kind of actions bad? And what sort of goodness enables sustaining exchange of altruism? We herein challenge this question with an evolutionary perspective. We generalize social reputation as 'Honor-score' (H-score) and examine the conditions under which individuals in a group stably maintain cooperative relationships based on indirect reciprocity. We examine the condition for evolutionarily stable strategies (ESSs) over 4096 possible cases exhaustively. Mathematical analysis reveals that only eight cases called 'leading eight' are crucial to the evolution of indirect reciprocity. Each in the leading eight shares two common characteristics: (i) cooperation with good persons is regarded as good while defection against them is regarded as bad, and (ii) defection against bad persons should be regarded as a good behavior because it works as sanction. Our results give one solution to the definition of goodness from an evolutionary viewpoint. In addition, we believe that the formalism of reputation dynamics gives general insights into the way social information is generated, handled, and transmitted in animal societies. PMID- 15363934 TI - Modeling the invasion of recessive Bt-resistant insects: an impact on transgenic plants. AB - There is a growing public concern on ecological and evolutionary consequence of the use of genetically modified organisms. We study the impact of Bt-resistant pests on genetically modified Bt crops. We develop and analyse a conceptual reaction-diffusion model of the Bt crop-Bt-susceptible insects-Bt-resistant insects to simulate the invasion of Bt-resistant insects. We show by means of computer simulations that there is a key parameter, which we define as the growth number that characterizes the insects' fitness. We also show that the Bt resistant insect invasion can lead to inhomogeneity in plant and insect spatial distributions. The plant biomass is found to be essentially dependent on the duration of the Bt-resistant insect reproduction period. There are two types of this dependence. One of them exhibits, respectively, higher plant biomass in comparison with another. The ambiguity in the response of the Bt crop-Bt susceptible insects system to the invasion of Bt-resistant insects can lead to serious complications in attempts to regulate the dynamics of the system. PMID- 15363935 TI - Mate choice when males are in patches: optimal strategies and good rules of thumb. AB - In standard mate-choice models, females encounter males sequentially and decide whether to inspect the quality of another male or to accept a male already inspected. What changes when males are clumped in patches and there is a significant cost to travel between patches? We use stochastic dynamic programming to derive optimum strategies under various assumptions. With zero costs to returning to a male in the current patch, the optimal strategy accepts males above a quality threshold which is constant whenever one or more males in the patch remain uninspected; this threshold drops when inspecting the last male in the patch, so returns may occur only then and are never to a male in a previously inspected patch. With non-zero within-patch return costs, such a two-threshold rule still performs extremely well, but a more gradual decline in acceptance threshold is optimal. Inability to return at all need not decrease performance by much. The acceptance threshold should also decline if it gets harder to discover the last males in a patch. Optimal strategies become more complex when mean male quality varies systematically between patches or years, and females estimate this in a Bayesian manner through inspecting male qualities. It can then be optimal to switch patch before inspecting all males on a patch, or, exceptionally, to return to an earlier patch. We compare performance of various rules of thumb in these environments and in ones without a patch structure. A two-threshold rule performs excellently, as do various simplifications of it. The best-of-N rule outperforms threshold rules only in non-patchy environments with between-year quality variation. The cutoff rule performs poorly. PMID- 15363936 TI - Resistance reversal action of ketoconazole against mefloquine resistance of Plasmodium yoelii nigeriensis. AB - Ketoconazole at 200 mg/kg dose has been found to exert marginal antimalarial action against multidrug resistant (MDR) Plasmodium yoelii nigeriensis (P. yoelii nigeriensis) in Swiss mice with 25% protection (2/8 mice) while at lower Ketoconazole dose i.e., 75-100 mg/kg, 14.28% mice were protected. Mefloquine (MFQ) (at 8 and 16 mg/kg) exerted suppressive action against MDR P. yoelii nigeriensis resulting in 25 and 14.28% protection of mice respectively. Combined treatment with Ketoconazole and mefloquine resulted in protection of 5/6 mice (83.33%) at MFQ 4 mg/kg + Ketoconazole 100 mg/kg dose, 7/8 (87.5%) mice at MFQ 8 mg/kg + Ketoconazole 20 mg/kg dose and 5/7 (71.42%) mice at MFQ 16 mg/kg + Ketoconazole 25 mg/kg dose and 5/6 (83.33%) mice at MFQ 16 mg/kg + Ketoconazole 100 mg/kg dose. Ketoconazole has been found to enhance the protective effect of mefloquine against MFQ resistant P. yoelii nigeriensis resulting in 66-88% protection of the mice treated with the appropriate combinations. The combination also increased suppression of parasitaemia at different times. The Ketoconazole combination with MFQ significantly increased the mean survival time of the treated mice compared to individual drugs alone. The study shows that Ketoconazole when administered with MFQ exerts bio-enhancing action against mefloquine resistance of MDR P. yoelii nigeriensis. PMID- 15363937 TI - Plasmodium yoelii: activity of azithromycin in combination with pyrimethamine or sulfadoxine against blood and sporozoite induced infections in Swiss mice. AB - The efficacy of pyrimethamine or sulfadoxine administered in combination with azithromycin was examined in a rodent malaria model. Outbred Swiss mice infected with blood stage parasites were treated from day 0 to day 3 and efficacy of different regimens was monitored in terms of the curative response and the delay time to reach 2% parasitaemia (2% DT). Administration of azithromycin alone at 60 mg/kg/day produced curative response while lower doses showed marginally delayed 2% DT. A marked potentiation in activities of pyrimethamine (100-fold) or sulfadoxine (10-fold) was observed when administered at non-curative doses of 0.1 mg/kg/day in combination with azithromycin (30 mg/kg/day) against blood stage parasites. A combination of 10 mg/kg/day azithromycin with 0.3 mg/kg/day sulfadoxine was also curative. Likewise in the causal prophylactic test, a combination regimen comprising 1/16th and 1/3rd the individual curative doses of pyrimethamine and azithromycin, respectively, prevented the development of patent infection after Plasmodium yoelii sporozoite challenge. Our results suggest that a combination of azithromycin with the second line treatment regimen of fansidar may enhance the therapeutic efficacy of the latter and also provide better prophylaxis against Plasmodium falciparum malaria. PMID- 15363938 TI - Trichomonas vaginalis: characterization of a 39-kDa cysteine proteinase found in patient vaginal secretions. AB - Trichomonosis, a chronic sexually transmitted disease, remains a public health problem affecting yearly over 170 million people worldwide. This disease is caused by Trichomonas vaginalis, a protozoan flagellate rich in cysteine proteinases (CPs). Although CPs are involved in trichomonal cytopathogenicity, only few of them have been defined as virulence factors. In this study, we characterize a T. vaginalis 39-kDa proteinase (CP39) found in vaginal secretions from patients with trichomonosis. The CP39 proteinase bound to HeLa epithelial cells, vaginal epithelial cells (VECs), and human prostatic cancer cells (DU 145). CP39 did not bind to a human colon cancer (CaCo) cell line, suggesting tissue-specific binding. CP39 was found in six fresh trichomonad isolates tested. In two-dimensional gels, CP39 appeared as a single spot with a pI 4.5. CP39 is inhibited by E-64, stable at 50 degrees C, and active in a wide pH range (3.6 9.0), with an optimum pH at 7.0. In addition, CP39 degraded collagens I, III, IV, and V, human fibronectin, human hemoglobin, and human immunoglobulins A and G. Indirect immunofluorescence detected CP39 on the parasite surface with specific polyclonal antibody to purified CP39. Finally, CP39 was found to be immunogenic, as evidenced by detection on immunoblots with serum of patients with trichomonosis, but not control individuals. These data suggest that CP39 may play a role during trichomonal infection. PMID- 15363939 TI - Characterization of the effect of retinol on Plasmodium falciparum in vitro. AB - A preliminary study from our laboratory found retinol (vitamin A alcohol) to have in vitro activity against Plasmodium falciparum at concentrations close to those in normal human serum (1-3 microM). To characterize the antimalarial potential of retinol in more detail, the 3D7 and K1 laboratory strains of P. falciparum were maintained in continuous culture and [3H]hypoxanthine incorporation and microscopy were used to assess the effect of retinol against asexual stages of the parasite life-cycle. Losses of retinol and retinol-associated hemolysis were also quantified in the in vitro culture system. There were retinol losses of >50% but no hemolysis was observed with added retinol concentrations up to 100 microM. All stages of parasite development showed comparable sensitivity to retinol including merozoite invasion (range of mean IC50 values 10.1-21.4 microM after adjustment for losses). Retinol pre-treatment of uninfected RBC did not inhibit merozoite invasion. Retinol treatment was associated with increased vacuolization within the parasite food vacuole and evidence of parasite membrane rupture. These appearances were similar to those seen with quinoline and artemisinin compounds. Although these data do not support a role for acute retinol supplementation in the treatment of falciparum malaria, they add to knowledge regarding potential antimalarial therapies and justify assessment of more potent synthetic retinoids and their metabolites. PMID- 15363940 TI - Onchocerca volvulus: expression and immunolocalization of a nematode cathepsin D like lysosomal aspartic protease. AB - The N-terminal region of the cathepsin D-like aspartic protease from the human filarial parasite Onchocerca volvulus was expressed as His-tag fusion protein. Light and electron microscopic immunohistology using antibodies against the recombinant protein showed labeling of lysosomes in the hypodermis and epithelia of the intestine and the reproductive organs of Onchocerca. While developing oocytes were negative, mature oocytes and early morulae showed strong labeling. In older embryos and mature microfilariae, stained lysosomes were only found in a few cells. Cell death in degenerating microfilariae of patients untreated and treated with microfilaricidal drugs was associated with strong expression of aspartic protease. IgG1, IgG4, and IgE antibodies reactive with the recombinant protein were demonstrated in sera from onchocerciasis patients indicating exposure and recognition of the enzyme by the host's defence system. The aspartic protease of O. volvulus appears to function in intestinal digestion and tissue degradation of the filaria. PMID- 15363941 TI - Plasmodium falciparum-infected erythrocytes bind to the RGD motif of fibronectin via the band 3-related adhesin. AB - Previously it was shown that Plasmodium falciparum-infected erythrocytes bound to thrombospondin by the interaction of the peptidic sequence, HPLQKTY, of the band 3 protein of infected erythrocytes, and the RGD motif of thrombospondin. Here, we show that falciparum-parasitized erythrocytes bind to immobilized fibronectin by the RGD sequence of fibronectin. Involvement of the HPLQKTY region of band 3 in binding was demonstrated by inhibition of adhesion of parasitized erythrocytes to fibronectin by an HPLQKTY-containing peptide and the binding of the HPLQKTY peptide to the RGD sequence of immobilized fibronectin. Since fibronectin occurs on endothelial cells and platelets, this interaction may contribute to the binding of falciparum-infected erythrocytes to such host cells. PMID- 15363942 TI - Echinococcus multilocularis: identification and molecular characterization of a Ral-like small GTP-binding protein. AB - In mammals, Ral (Ras-like) GTPases have been implicated in the regulation of several cellular key processes such as oncogenic transformation, endocytosis, and actin-cytoskeleton dynamics. Here we provide, for the first time, molecular data on a Ral homologue from a parasitic helminth. We have cloned and characterized the complete cDNA molecule and the chromosomal locus encoding a novel GTP binding protein, EmRal, of the human parasite Echinococcus multilocularis. The encoded protein contained all highly conserved amino acid residues of the protein family at corresponding positions and shared significant sequence homologies with human RalA (53% identity) and RalB (54%). Upon heterologous expression of EmRal in Escherichia coli, the recombinant protein was able to bind GTP, thus indicating functionality of the Echinococcus factor. Using an in vitro prenylation assay, the purified protein was shown to be geranylgernylated, but not farnesylated, in both rabbit reticulocyte and Echinococcus cell extracts. The EmRal mRNA was found to be processed via trans-splicing and, using RT-PCR and virtual Northern blot experiments, expression of the factor could be demonstrated for the larval stages metacestode and protoscolex during an infection of the intermediate host. The data presented herein provide a solid basis for further investigations on Ras-Ral signaling mechanisms in Echinococcus. PMID- 15363943 TI - Leishmania (Leishmania) amazonensis: purification and characterization of a promastigote serine protease. AB - Pathogenic protozoan proteases play crucial roles in the host-parasite interaction, and its characterization contributes to the understanding of protozoan disease mechanisms. A Leishmania amazonensis promastigote protease was purified 36-fold, using aprotinin-agarose affinity chromatography and gel filtration high performance liquid chromatography, yielding a total recovery of 49%. The molecular mass of active enzyme obtained from native gel filtration HPLC and SDS-PAGE under conditions of reduction and non-reduction was 68 kDa, suggesting that the enzyme may exist as a monomer. The protease isoelectric point (pI) was around 4.45 and, as demonstrated by deglycosylation assay, it did not have any carbohydrate content. The optimal pH and temperature of the enzyme were 8.0 and 28 degrees C, respectively, determined using alpha-N-rho-tosyl-L-arginyl methyl ester (L-TAME) as substrate. Assays of thermal stability indicated that 50% of the enzymatic activity was preserved after 4 min of pre-treatment at 42 degrees C and after 24 h of pre-treatment at 37 degrees C, both in the absence of substrate. Hemoglobin, bovine serum albumin (BSA), ovalbumin, and both gelatin and peptide substrates containing arginine in ester bound were hydrolyzed by 68 kDa protease. The insulin beta-chain was also hydrolyzed by the protease, and four peptidic bonds (L11-V12, E13-A14, L15-Y16, and Y16-L17) were susceptible to the 68-kDa protease action. Inhibition studies suggested that the enzyme belonged to a serine protease class inhibited by calcium ions and activated by manganese ions. These findings demonstrate that the L. amazonensis 68-kDa serine protease differs from those of other protozoan parasites. PMID- 15363944 TI - Plasmodium falciparum: adherence of the parasite-infected erythrocytes to chondroitin sulfate proteoglycans bearing structurally distinct chondroitin sulfate chains. AB - Infection with Plasmodium falciparum during pregnancy leads to the selective adherence of infected red blood cells (IRBCs) in the placenta causing placental malaria. The IRBC adherence is mediated through the chondroitin 4-sulfate (C4S) chains of unusually low-sulfated chondroitin sulfate proteoglycans (CSPGs) in the placenta. To study the structural interactions involved in C4S-IRBC adherence, various investigators have used CSPGs from different sources. Since the structural characteristics of the polysaccharide chains in CSPGs from various sources differ substantially, the CSPGs are likely to differentially bind IRBCs. In this study, the CSPG purified from bovine trachea, a CSPG form of human recombinant thrombomodulin (TM-CSPG), two CSPG fractions from bovine cornea, and the CSPGs of human placenta, the natural receptor, were studied in parallel for their IRBC binding characteristics. The TM-CSPG and corneal CSPG fractions could bind IRBCs at significantly higher density compared to the placental CSPGs. However, the avidity of IRBC binding by TM-CSPG was considerably low compared to placental CSPGs. The corneal CSPGs have substantially higher binding strengths. The bovine tracheal CSPG bound IRBCs at much lower density and exhibited significantly lower avidity than the placental CSPGs. These data demonstrated that the bovine tracheal CSPG and TM-CSPG are not ideal for studying the fine structural interactions involved in the IRBC adherence to the placental C4S, whereas the bovine corneal CSPGs are better alternatives to the placental CSPGs for determining these interactions. PMID- 15363945 TI - The modelling of mononuclear phagocyte-connective tissue adhesion in vitro: application to disclose a specific inhibitory effect of Leishmania infection. AB - In this work, we have developed an adhesion assay to study interactions between mononuclear phagocytes and connective tissue in vitro and show its potential use to study diseases caused by intracellular microorganisms. The assay reproduces most of the characteristics of macrophage adhesion to connective tissue in vivo, such as: preferential adhesion to inflamed connective tissue, divalent cation and integrin dependence, and up-regulation upon cell activation. The phagocyte adhesion to connective tissue was inhibited by infection with Leishmania (58+/ 22%, p < 0.05) and was not affected by infection with Mycobacterium or by endocytosis of latex beads. Manganese partially reverted the loss in adherence produced by Leishmania infection, indicating that the mechanisms regulating the function of integrins are affected by cell infection with Leishmania. This assay might be a useful tool for the study of the mechanisms by which mononuclear phagocytes play a role in the immune-inflammatory response and in the development of lesions. PMID- 15363946 TI - Receptor imaging in the thorax with PET. AB - This review focuses on positron emission tomography (PET)-imaging of receptors in the sympathetic and the parasympathetic systems of heart and lung and highlights the human applications of PET. For the alpha-adrenoceptor, only [11C]GB67 (N2-[6 [(4-amino-6,7-dimethoxy-2-quinazolinyl)(methyl)amino]hexyl]-N2-[11C]methyl-2 furamide hydrochloride) has been developed. Its potential for application in patients needs to be assessed. For both the beta-adrenergic and the muscarinic systems, potent PET radioligands have been prepared and evaluated in patients. It has been possible to measure receptor densities quantitatively in human heart [[11C]MQNB: [11C]methylquinuclidinyl benzilate, [11C]CGP12177: S-(3'-t-butylamino 2'-hydroxypropoxy)-benzimidazol-2-[11C]one and [11C]CGP12388: (S)-4-(3-(2' [11C]isopropylamino)-2-hydroxypropoxy)-2H-benzimidazol-2-one] and qualitatively in lung [[11C]VC002: N-[11C]-methyl-piperidin-4-yl-2-cyclohexyl-2-hydroxy-2 phenylacetate and [11C]CGP12177]. Besides these subtype nonselective radioligands, the development of compounds that are selective for one subtype are ongoing and have not found successful application in humans yet. PMID- 15363947 TI - Neuroendocrinology of gastric H+ and duodenal HCO3- secretion: the role of brain gut axis. AB - Gastric H+ and duodenal HCO3- secretions are precisely regulated by neuro hormonal mechanisms at central and peripheral levels to match the rate of these secretions with the type of stimulation of sensory receptors in the head area (sight, smell, taste, etc.) and in the gastro-intestinal system. Two-way communication pathways operate between the brain and the gut, each comprising afferent fibers signaling sensory information from the gut to the brain and efferent fibers transmitting signals in opposite direction. Short intramural and long extramural reflexes are triggered as well as various gut hormones are released by feeding that "cooperate" with the "brain-gut axis" in the alteration of exocrine and endocrine gastro-duodenal secretion, motility and blood circulation. The malfunction of gastric or duodenal secretory mechanisms may lead to disturbances of gastric H+-pepsin or duodenal mucus-HCO3- secretion and to gastro-duodenal disorders and diseases. This review presents recent advances in pathophysiological mechanisms underlying gastro-duodenal secretory disorders. PMID- 15363948 TI - Propofol scavenges reactive oxygen species and inhibits the protein nitration induced by activated polymorphonuclear neutrophils. AB - Activated polymorphonuclear neutrophils may damage tissues through the release of biochemical mediators. Among them, peroxynitrite is responsible for hydroxylation reactions and nitration of proteins, or is metabolised into nitrate. We investigated the effect of propofol on the production of reactive oxygen species, the nitration of proteins and the formation of nitrate by activated human polymorphonuclear neutrophils. Propofol dose-dependently inhibited chemiluminescence, nitration of proteins and nitrate production in a concentration range from 10(-3) to 10(-6) mM. A significant correlation was observed between the logarithm of propofol concentration and the intensity of chemiluminescence (r2=0.90), the nitration of proteins (r2=0.67) and the production of nitrate (r2=0.79). Those results are consistent with the scavenging effect of propofol on peroxynitrite and could confer a protective property to propofol in pathological situations involving polymorphonuclear neutrophils activation. PMID- 15363949 TI - The respective N-hydroxypyrazole analogues of the classical glutamate receptor ligands ibotenic acid and (RS)-2-amino-2-(3-hydroxy-5-methyl-4-isoxazolyl)acetic acid. AB - We have determined the pharmacological activity of N-hydroxypyrazole analogues (3a and 4a) of the classical glutamate receptor ligands ibotenic acid and (RS)-2 amino-2-(3-hydroxy-5-methyl-4-isoxazolyl)acetic acid (AMAA), as well as substituted derivatives of these two compounds. The pharmacological profile of 3a is closer to that of thioibotenic acid rather than ibotenic acid, while 4a is a selective N-methyl-D-aspartic acid (NMDA) receptor agonist. Ring substitution of 3a and 4a leads to NMDA receptor antagonists. Whereas efficacy of 3a derivatives at mglu2 receptor decreases from agonism via partial agonism to antagonism with increasing substituent size, substitution abolishes affinity for mglu1 and mglu4 receptors. Ligand- and receptor-based modelling approaches assist in explaining these pharmacological trends among the metabotropic receptors and suggest a mechanism of partial agonism at mglu2 receptor similar to that proposed for the GluR2 glutamate receptor. PMID- 15363950 TI - Human organic cation transporter 3 mediates the transport of antiarrhythmic drugs. AB - Antiarrhythmic drugs have been considered to be transported by the organic cation transport system. The purpose of this study was to elucidate the molecular mechanism underlying the transport of antiarrhythmic drugs using cells from the second segment of the proximal tubule (S2) cells of mice expressing human-organic cation transporter 3 (S2 human-OCT3). The antiarrhythmic drugs tested were cibenzoline, disopyramide, lidocaine, mexiletine, phenytoin, pilsicanide, procainamide and quinidine. Human-OCT3 mediated a time- and dose-dependent uptake of quinidine and lidocaine, with Km values of 216 and 139 microM, respectively. Human-OCT3 also mediated the uptake of disopyramide and procainamide but not that of phenytoin. All antiarrhythmic drugs tested inhibited histamine uptake mediated by human-OCT3 in a dose-dependent manner. The IC50 values of antiarrhythmic drugs for human-OCT3 ranged between 0.75 and 656 microM. Kinetic analysis revealed that disopyramide, lidocaine, procainamide and quinidine inhibited histamine uptake mediated by human-OCT3 in a competitive manner. In conclusion, these results suggest that human-OCT3 mediates the transport of antiarrhythmic drugs, which may be the mechanism underlying the distribution and the elimination of these drugs. PMID- 15363951 TI - Exploring the pharmacology of the leukotriene B4 receptor BLT1, without the confounding effects of BLT2. AB - Most previous studies of leukotriene B4 (LTB4) pharmacology using primary leukocyte cultures and myeloid cell lines do not differentiate between leukotriene BLT1 and BLT2 receptor activation because both receptors are often expressed by these cells. Here we show that in HeLa cells expressing BLT1 but not BLT2 receptors, BLT1 receptor activation resulted in IP3 mediated calcium release from intracellular stores initially, followed by calcium influx through cell membrane channels. BLT1 calcium signalling was sensitive to the activity of protein kinase C (PKC), protein kinase A (PKA) and protein-tyrosine kinases (PTKs), as well as changes in membrane cholesterol levels and treatments that are known to disrupt normal membrane physiology and/or lipid rafts. Inhibition of MAP kinases, Rho-associated kinases, or phosphoinositol-3-kinases (PI3K) had no effect on BLT1 receptor induced calcium signalling, and the receptor was insensitive to the redox state of the extracellular compartment. PMID- 15363952 TI - In vitro characterization of AR-A000002, a novel 5-hydroxytryptamine(1B) autoreceptor antagonist. AB - The in vitro pharmacological properties of AR-A000002 ((R)-N-[5-methyl-8-(4 methylpiperazin-1-yl)-1,2,3,4-tetrahydro-2-naphthyl]-4-morpholinobenzamide), a novel 5-hydroxytryptamine (5-HT)(1B) receptor antagonist, were studied. AR A000002 bound with high affinity to guinea pig cortex and recombinant guinea pig 5-HT(1B) receptors (Ki=0.24 and 0.47 nM) and with 10-fold lower affinity to 5 HT(1D) receptors. The compound displayed weak or no affinity for 63 other binding sites tested. In [35S]GTPgammaS assays AR-A000002 showed 50% efficacy and inhibited 5-HT stimulation with 66% and a pA2 value of 8.9. In slices of guinea pig cortex, AR-A000002 enhanced the outflow of [3H]5-HT upon electrical stimulation. The compound blocked sumatriptan-evoked contraction of rabbit saphenous veins without inducing any contraction itself. Thus, in these two systems AR-A000002 behaved as a 5-HT(1B) receptor antagonist. It is concluded that AR-A000002 is a selective high affinity 5HT(1B) receptor ligand that shows partial agonist activity in recombinant systems. In native tissues AR-A000002 behaves as a 5-HT(1B) receptor antagonist. PMID- 15363953 TI - Pharmacological characterization and radioligand binding properties of a high affinity, nonpeptide, bradykinin B1 receptor antagonist. AB - Compound A (N-[2-[4-(4,5-dihydro-1H-imidazol-2-yl)phenyl]ethyl]-2-[(2R)-1-(2 napthylsulfonyl)-3-oxo-1,2,3,4-tetrahydroquinoxalin-2-yl]acetamide) is a member of a new class of aryl sulfonamide dihydroquinoxalinone bradykinin B1 receptor antagonists that should be useful pharmacological tools. Here we report on some of the pharmacological properties of compound A as well as the characterization of [35S]compound A as the first nonpeptide bradykinin B1 receptor radioligand. Compound A inhibited tritiated peptide ligand binding to the cloned human, rabbit, dog, and rat bradykinin B1 receptors expressed in CHO cells with Ki values of 0.016, 0.050, 0.56, and 29 nM, respectively. It was inactive at 10 microM in binding assays with the cloned human bradykinin B2 receptor. In functional antagonist assays with the cloned bradykinin B1 receptors, compound A inhibited agonist-induced signaling with activities consistent with the competition binding results, but had no antagonist activity at the bradykinin B2 receptor. Compound A was also found to be a potent antagonist in a rabbit aorta tissue bath preparation and to effectively block des-Arg9 bradykinin depressor responses in lipopolysaccharide-treated rabbit following intravenous administration. The binding of [35S]compound A was evaluated with the cloned bradykinin B1 receptors. In assays with human, rabbit, and dog receptors, [35S]compound A labeled a single site with Kd values of 0.012, 0.064, and 0.37 nM, respectively, and with binding site densities equivalent to those obtained using the conventional tritiated peptide ligands. Binding assays with the cloned rat bradykinin B1 receptor were not successful, presumably due to the low affinity of the ligand for this species receptor. There was no specific binding of the ligand detected in CHO cells expressing the human bradykinin B2 receptor. In assays with the cloned human bradykinin B1 receptor, the pharmacologies of the binding of [35S]compound A and [3H][Leu9]des-Arg10-kallidin were the same. The high signal-to-noise ratio obtained with [35S]compound A will allow this ligand to be a very useful tool for future investigations of the bradykinin B1 receptor. PMID- 15363954 TI - Ferulic acid inhibits vascular smooth muscle cell proliferation induced by angiotensin II. AB - The aim of this study was to determine the effects of ferulic acid on the proliferation and molecular mechanism in cultured vascular smooth muscle cell (VSMC) induced by angiotensin II. It was shown that ferulic acid significantly inhibited angiotensin II-induced VSMC proliferation in a dose-dependent manner. Western blotting analyses suggest that the antiproliferative effect of ferulic acid was involved in the mitogen-activated protein kinases (MAPKs) pathway. While no effect on p38, ferulic acid markedly inactivated the extracellular signal regulated kinases (ERK1/2) and c-Jun N-terminal kinases (JNK), indicating that the inhibition of ferulic acid on VSMC proliferation was associated with ERK1/2 and JNK rather than p38 pathway. On the expression of cell cycle regulatory proteins, ferulic acid elevated the protein content of p21(waf1/cip1), decreased expression of cyclin D1 and inhibited phosphorylation of retinoblastoma protein, suggesting that ferulic acid inhibited VSMC proliferation by regulating the cell progression from G1 to S phase. The inactivation of MAPKs and modulation of cell cycle proteins of ferulic acid may be of importance in preventing cardiovascular disease. PMID- 15363955 TI - Allosteric modulation and constitutive activity of fusion proteins between the adenosine A1 receptor and different 351Cys-mutated Gi alpha-subunits. AB - We studied fusion proteins between the human adenosine A1 receptor and different 351Cys-mutated G(i1) alpha-subunits (A1-Gialpha) with respect to two important concepts in receptor pharmacology, i.e. allosteric modulation and constitutive activity/inverse agonism. The aim of our study was twofold. We first analysed whether such fusion products are still subject to allosteric modulation, and, secondly, we investigated the potential utility of the fusion proteins to study constitutive receptor activity. We determined the pharmacological profile of nine different A1-Gialpha fusion proteins in radioligand binding studies. In addition, we performed [35S]GTPgammaS binding experiments to study receptor and G protein activation of selected A1-Gialpha fusion proteins. Compared to unfused adenosine A1 receptors, the affinity of N6-cyclopentyladenosine (CPA) at wild-type A1 Gialpha fusion proteins (351Cys) increased more than eightfold, while the affinity of 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) did not change significantly. Furthermore, we showed that the allosteric enhancer of agonist binding, PD81,723 (2-amino-4,5-dimethyl-3-thienyl-[3-(trifluoromethyl) phenyl]methanone), elicited similar effects on ligand binding; i.e. CPA binding to the A1-Gialpha fusion proteins was enhanced, whereas the affinity of DPCPX was hardly affected. Moreover, sodium ions were unable to decrease agonist binding to the majority of the A1-Gialpha fusion proteins, presumably because they exhibit their effect through uncoupling of the R-G complex. From [35S]GTPgammaS binding experiments, we learned that all the A1-Gialpha fusion proteins tested had a higher basal receptor activity than the unfused adenosine A1 receptor, thereby providing improved conditions to observe inverse agonism. Moreover, the maximal CPA-induced stimulation of basal [35S]GTPgammaS binding was increased for the five A1-Gialpha fusion proteins tested, whereas the inhibition induced by 8 cyclopentyltheophylline (CPT) was more pronounced at 351Cys, 351Ile, and 351Val A1-Gialpha fusion proteins. Thus, the maximal receptor (de)activation depended on the amino acid at position 351 of the Gi alpha-subunit. In conclusion, A1-Gialpha fusion proteins, especially with 351Cys and 351Ile, can be used as research tools to investigate inverse agonism, due to their increased readout window in [35S]GTPgammaS binding experiments. PMID- 15363956 TI - Brain phospholipase C-diacylglycerol lipase pathway is involved in vasopressin induced release of noradrenaline and adrenaline from adrenal medulla in rats. AB - Recently, we reported that intracerebroventricularly (i.c.v.) administered arginine-vasopressin evokes the release of noradrenaline and adrenaline from adrenal medulla by brain thromboxane A2-mediated mechanisms in rats. These results suggest the involvement of brain arachidonic acid in the vasopressin induced activation of the central adrenomedullary outflow. Arachidonic acid is released mainly by two pathways: phospholipase A2 (PLA2)-dependent pathway; phospholipase C (PLC)- and diacylglycerol lipase-dependent pathway. In the present study, therefore, we attempted to identify which pathway is involved in the vasopressin-induced release of both catecholamines from adrenal medulla using urethane-anesthetized rats. Vasopressin (0.2 nmol/animal, i.c.v.)-induced elevation of plasma noradrenaline and adrenaline was dose-dependently reduced by neomycin [0.28 and 0.55 micromol (250 and 500 microg)/animal, i.c.v.] and 1-[6 [[(17beta)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U-73122) [5 and 10 nmol (2.3 and 4.6 microg)/animal, i.c.v.] (inhibitors of PLC), and also by 1,6-bis(cyclohexyloximinocarbonylamino)hexane (RHC-80267) [1.3 and 2.6 micromol (500 and 1000 microg)/animal, i.c.v.] (an inhibitor of diacylglycerol lipase). On the other hand, mepacrine [1.1 and 2.2 micromol (500 and 1000 microg)/animal, i.c.v.] (an inhibitor of PLA2) was largely ineffective on the vasopressin-induced elevation of plasma catecholamines. These results suggest that vasopressin evokes the release of noradrenaline and adrenaline from adrenal medulla by the brain PLC- and diacylglycerol lipase-dependent mechanisms in rats. PMID- 15363957 TI - Characterization of the complex morphinan derivative BU72 as a high efficacy, long-lasting mu-opioid receptor agonist. AB - The development of buprenorphine as a treatment for opiate abuse and dependence has drawn attention to opioid ligands that have agonist actions followed by long lasting antagonist actions. In a search for alternatives to buprenorphine, we discovered a bridged pyrrolidinomorphinan (BU72). In vitro, BU72 displayed high affinity and efficacy for mu-opioid receptors, but was also a partial delta opioid receptor agonist and a full kappa-opioid receptor agonist. BU72 was a highly potent and long-lasting antinociceptive agent against both thermal and chemical nociception in the mouse and against thermal nociception in the monkey. These effects were prevented by mu-, but not kappa- or delta-, opioid receptor antagonists. Once the agonist effects of BU72 had subsided, the compound acted to attenuate the antinociceptive action of morphine. BU72 is too efficacious for human use but manipulation to reduce efficacy could provide a lead to the development of a treatment for opioid dependence. PMID- 15363958 TI - Convulsant and anticonvulsant effects of bupropion in mice. AB - This study demonstrated that bupropion hydrochloride, an effective antidepressant and a commonly used smoking cessation aid, dose-dependently caused clonic convulsions in mice, with the CD50 (convulsive dose50, i.e., the dose producing convulsions in 50% of mice) at 119.7 mg kg(-1). An evaluation for anticonvulsant effects showed that bupropion in the doses of 15-30 mg kg(-1) protected against convulsions induced by maximal electroshock with the ED50 (effective dose50, i.e., the dose protected 50% of mice against convulsions) being 19.4 mg kg(-1). Bupropion had no effect on pentylenetetrazole- and kainic acid-induced convulsions. It is possible that the anticonvulsant activity of bupropion may be exploited for use in the treatment of epilepsy but it requires further investigations. PMID- 15363959 TI - Antagonism at metabotropic glutamate 5 receptors inhibits nicotine- and cocaine taking behaviours and prevents nicotine-triggered relapse to nicotine-seeking. AB - Previous studies in metabotropic glutamate 5 receptor (mGlu5 receptor) deficient mice have indicated the importance of this receptor in the self-administration of cocaine and locomotor sensitisation to this stimulant. Both ionotropic and metabotropic receptors have been implicated in drug-seeking and drug-taking behaviours, but the specific role of each subtype of metabotropic glutamate receptors (mGlu receptors) is still unknown. In the present series of experiments we further investigated the role of mGlu5 receptors on nicotine, cocaine- and food-taking behaviour. We also investigated the effects of the mGlu5 receptor antagonist MPEP (2-methyl-6-(phenylethynyl)pyridine) on the acute locomotor activating effects of nicotine, the expression of sensitisation to its repeated, intermittent administration, and nicotine-triggered relapse to nicotine-seeking behaviour. The results indicate that MPEP treatment reduced nicotine-induced drug seeking behaviour in a model of nicotine-triggered relapse to nicotine seeking. Furthermore, MPEP decreased both nicotine and cocaine self-administration without affecting food self-administration under similar schedules of reinforcement. Finally, MPEP reduced both the acute locomotor stimulant effects of nicotine as well as the expression of behavioural sensitisation to its repeated administration. Although the intravenous administration of MPEP at 1 and 10 mg/kg transiently reduced spontaneous locomotor activity during the first 25 min post administration, we also demonstrated that performance on the accelerating rotarod was not affected when MPEP was given 5 and 30 min prior to the test. Altogether, the present findings strengthen the hypothesis that selective antagonism at mGlu5 receptors may be a new potential pharmacotherapeutic approach for the treatment of drug dependence and addiction. PMID- 15363960 TI - Antidepressant-like activity of corticotropin-releasing factor type-1 receptor antagonists in mice. AB - The development of selective corticotropin-releasing factor type-1 (CRF1) receptor antagonists represents a potential novel treatment for depression. These studies evaluated CRF1 receptor antagonists for antidepressant-like activity in mice. Subchronic dosing of both R 121919 (3-[6-(dimethylamino)-4-methyl-pyrid-3 yl]-2,5-dimethyl-N,N-dipropyl-pyrazolo[2,3-a]pyrimidin-7-amine) and DMP 696 (4 (1,3-dimethoxyprop-2-ylamino)-2,7-dimethyl-8-(2,4-dichlorophenyl)-pyrazolo[1,5-a] 1,3,5-triazine) significantly decreased immobility time in the tail suspension test (at 30 and at 3 and 10 mg/kg, i.p., respectively). These antidepressant-like effects were observed at doses that did not impair general locomotor activity. Neither antalarmin (N-butyl-N-ethyl-[2,5,6-trimethyl-7-(2,4,6)trimethylphenyl)-7H pyrrolo[2,3-d]pyrimidin-4-yl]amine) nor DMP 904 (4-(3-pentylamino)-2,7-dimethyl-8 (2-methyl-4-methoxyphenyl)-pyrazolo-[1,5-a]-pyrimidine) had an effect indicative of antidepressant-like activity. These results suggest that the tail suspension assay may have utility to identify CRF1 receptor antagonists with antidepressant like activity. Moreover, the results lend support to the theory that some nonpeptidic CRF1 receptor antagonists may possess antidepressant-like activity and therefore represent a promising novel pharmacotherapeutic strategy in the treatment of depression. PMID- 15363961 TI - Adenosine A1 receptors modulate the anxiolytic-like effect of ethanol in the elevated plus-maze in mice. AB - The anxiolytic property of ethanol is generally accepted to be an important motivational factor for its consumption and the development of alcohol dependence. Recent studies suggest that adenosine receptors mediate important actions of ethanol, such as motor incoordination and hypnotic effects. In addition, several lines of evidence support the involvement of adenosine in anxiety. The aim of the present study was to evaluate the role of adenosine receptors in the anxiolytic-like effect of ethanol in mice. The effects of acute administration of the adenosine receptor antagonists caffeine (nonselective), 8 cyclopentyl-1,3-dipropylxanthine (DPCPX, adenosine A1 receptor antagonist) and 4 (2-[7-amino-2-[2-furyl][1,2,4]triazolo-[2,3-a][1,3,5]triazin-5-yl amino]ethyl)phenol (ZM241385, adenosine A(2A) receptor antagonist), together with the adenosine A1 receptor agonist 2-chloro-N6-cyclopentyladenosine (CCPA), and their interaction with ethanol in the elevated plus-maze test in mice were studied. The highest doses of caffeine (30.0 mg/kg, i.p.) and DPCPX (6.0 mg/kg, i.p.) produced an anxiogenic-like effect, while CCPA administration (0.25 mg/kg, i.p.) showed an anxiolytic-like activity. The prior administration of "non anxiogenic" doses of caffeine (10.0 mg/kg, i.p.) and DPCPX (3.0 mg/kg, i.p.), but not ZM241385 (1.0 mg/kg, i.p.), significantly reduced the anxiolytic-like effect of ethanol (1.2 g/kg, i.p.). Moreover, anxiolytic-like response was observed by the co-administration of "non-anxiolytic" doses of CCPA (0.125 mg/kg) and ethanol (0.6 g/kg). These results reinforce the involvement of adenosine in anxiety and suggest that the activation of adenosine A1 receptors, but not adenosine A(2A) receptors, mediate the anxiolytic-like effect induced by ethanol in mice. PMID- 15363962 TI - NMDA receptors and associated signaling pathways: a role in knee joint blood flow regulation. AB - Blood flow changes in response to N-methyl-D-aspartate (NMDA) receptor activation were assessed using a laser Doppler flowmeter. Treatment of the joint with NMDA (1 mM; 0.1 ml) resulted in a significant increase in blood flow while the control phosphate buffer (PB) injection (0.1 M; pH 7.4) had no effect. Blocking NMDA receptors with the antagonist MK 801 (0.1 mM) prevented the increase in blood flow observed following NMDA injection, suggesting specificity of action. The NMDA-evoked vasodilation has been shown to be mediated through activation of several intracellular signaling transduction molecules, namely nitric oxide, release of calcitonin gene-related peptide (CGRP) and CAM kinase II. Blocking actions of these molecules with L-NAME (10 mg/ml), CGRP(8-37) (0.01 mM) and KN-93 (1 microM), respectively, prevented the increase in blood flow induced by NMDA in the present study. These results provide new evidence implicating NMDA receptors in knee joint inflammatory responses. PMID- 15363963 TI - Intravenous atenolol and esmolol maintain the protective effect of ischemic preconditioning in vivo. AB - Catecholamines bind to alpha- and beta-adrenoreceptors and are capable of preconditioning ischemic myocardium. Our purpose was to investigate the effect of acute either short or prolonged i.v. administration of beta-adrenoreceptor antagonists on ischemic preconditioning in vivo. Fifty-five anesthetized rabbits were divided into 10 groups (n=5-7 per group) and were subjected to 30-min regional ischemia of the heart after ligation of a prominent left coronary artery and 3-h reperfusion after releasing the snare. Ischemic preconditioning was obtained by three cycles of 5-min ischemia separated by 10-min reperfusion. beta Adrenoreceptor blockade was obtained by the long acting beta-adrenoreceptor antagonist atenolol or by the short acting esmolol, which were given as a short 5 min infusion or as a prolonged 45-min infusion, starting respectively 20 min before and ending 15 min before the beginning of sustained ischemia, or starting 45 min before and ending immediately before the beginning of sustained ischemia. Atenolol was given at a rate of 0.2 mg min(-1) during 5 min or at a rate of 0.088 mg min(-1) as a 45-min infusion. Esmolol was given as an initial dose of 500 microg kg(-1) within 1 min, followed by a 4-min infusion at a rate of 50 microg kg(-1) min(-1) or as an initial dose of 3.4 mg within 1 min, followed by a 44-min infusion at a rate of 0.15 mg min(-1). Blood pressure and heart rate were continuously monitored. The infarcted and risk areas were delineated with the aid of tetrazolium chloride staining and fluorescent Zn-Cd particles. Infarct size was expressed in percent of the area at risk. All the animals without preconditioning developed an infarct size ranging between 36.3+/-2.4% and 49.6+/ 7.6% (P=NS) and all the preconditioning groups developed an infarct size ranging between 14.9+/-1.2% and 21.0+/-2.2% (P=NS). All the preconditioning groups, independently of the use of beta-adrenoreceptor antagonists, had a smaller infarct size than the control group, which developed an infarct size of 47.3+/ 2.5% (P<0.01). Intravenous atenolol and esmolol, independent of timing and mode of administration, does not seem to interfere with protection afforded by ischemic preconditioning in vivo. PMID- 15363964 TI - Mechanisms of hydroxyl radical-induced contraction of rat aorta. AB - The present study was designed to investigate the effects of hydroxyl radicals (*OH), generated via the Fe2+-mediated Fenton reaction, on isolated rat aortic rings with and without endothelium. In the absence of any vasoactive agent, generation of *OH alone elicited an endothelium-independent contraction in rat aortic rings in a concentration-dependent manner. Hydroxyl radical-induced contractions of denuded rat aortic rings appeared, however, to be slightly stronger than those on intact rat aortic rings. The contractile responses to *OH were neither reversible nor reproducible in the same ring; even small concentrations of *OH radicals resulted in tachyphylaxis. Removal of extracellular calcium ions (Ca2+) or buffering intracellular Ca2+ with 10 microM acetyl methyl ester of bis(o-aminophenoxy) ethane-N,N,N',N',-tetraacetic acid (BAPTA-AM) significantly attenuated the contractile actions of *OH radicals. The presence of 1 microM staurosporine, 1 microM bisindolylmaleimide I, 1 microM Go6976 [inhibitor of protein kinase C (PKC)], 2 microM PD-980592 (inhibitor of ERK), 10 microM genistein, and 1 microM wortmannin significantly inhibited the contractions induced by *OH. Proadifen (10 microM), on the other hand, significantly potentiated the hydroxyl radical-induced contractions. Exposure of primary cultured aortic smooth muscle cells to *OH produced significant, rapid rises of intracellular free Ca2+ ([Ca2+]i). Several, specific antagonists of possible endogenously formed vasoconstrictors did not inhibit or attenuate either hydroxyl radical-induced contractions or the elevation of [Ca2+]i. Our new results suggest that hydroxyl radical-triggered contractions on rat aortic rings are Ca2+-dependent. Several intracellular signal transduction systems seem to play some role in hydroxyl radical-induced vasoconstriction of rat aortic rings. PMID- 15363965 TI - Cardioprotective effect of MCC-135 is associated with inhibition of Ca2+ overload in ischemic/reperfused hearts. AB - Calcium (Ca2+) overload is an important pathophysiological factor in myocardial ischemic/reperfusion injury. We investigated the effects of a cardioprotective drug, MCC-135, 5-methyl-2-(1-piperazinyl) benzenesulfonic acid monohydrate, on (1) cardiac contractile dysfunction and Ca2+ overload induced by ischemia and reperfusion, and (2) the Na+/Ca2+ exchanger in Langendorff-perfused rat hearts. Low-flow 45-min ischemia and 30-min reperfusion decreased developed tension and increased ventricular Ca2+ content, effects which were ameliorated by MCC-135 and amiloride given after reperfusion. Combination of intracellular Na+ overload induced by monensin (Na+ ionophore; 5 microM) and zero-flow 15-min ischemia followed by 30-min reperfusion resulted in a decrease in developed tension and in the intracellular Na+-dependent increase in ventricular Ca2+ content. MCC-135 and the highest dose of amiloride given after reperfusion reduced the increase in ventricular Ca2+ content, whereas developed tension was increased only with MCC 135. These results suggest that the cardioprotective effect of MCC-135 in ischemia/reperfusion is associated with suppression of Ca2+ overload and is attributable to inhibition of intracellular Na+-dependent Ca2+ influx via the Na+/Ca2+ exchanger. PMID- 15363966 TI - Vasoconstrictor effects of 8-iso-prostaglandin E2 and 8-iso-prostaglandin F(2alpha) on human umbilical vein. AB - The present study was undertaken to determine whether 8-iso-prostaglandin E2 and 8-iso-prostaglandin F(2alpha) posses contractile action on human umbilical vein and to evaluate the possible involvement of prostanoid TP receptors in this effect. Human umbilical vein rings were mounted in organ baths and concentration response curves to 8-iso-prostaglandin E2 or 8-iso-prostaglandin F(2alpha) were constructed. Both isoprostanes evoked concentration-dependent contraction. 8-iso prostaglandin E2 (pEC50=6.90+/-0.03) was significantly more potent than 8-iso prostaglandin F(2alpha) (pEC50=6.10+/-0.04). However, both isoprostanes were equieffective. The prostanoid TP receptor antagonists, ICI-192,605 (4-(Z)-6-(2-o Chlorophenyl-4-o-hydroxyphenyl-1,3-dioxan-cis-5-yl)hexenoic acid) and SQ-29548 (7 [3-[[2-[(phenylamino)carbonyl]hydrazino]methyl]-7-oxabicyclo[2.2.1]hept-2-yl] [1S(1alpha,2alpha(Z),3alpha,4alpha)]-5-Heptenoic acid) produced a competitive rightward shift of 8-iso-prostaglandin E2 concentration-response curves with pKB values of 8.91+/-0.04 and 8.07+/-0.07, respectively. When ICI-192,605 (1 nM) and SQ-29548 (10 nM) were evaluated against 8-iso-prostaglandin F(2alpha) they produced a parallel rightward displacement of 8-iso-prostaglandin F(2alpha) concentration-response curves without affecting the maximum responses giving pA2 values of 9.02+/-0.12 and 8.26+/-0.13, respectively. In conclusion, the present study describes for the first time the vasoconstrictor action of 8-iso prostaglandin E2 and 8-iso-prostaglandin F(2alpha) in human umbilical vein. Furthermore, the affinity values obtained with ICI-192,605 and SQ-29548 provide strong pharmacological evidence of prostanoid TP receptors involvement in this effect. PMID- 15363967 TI - Delayed exercise-induced protection against arrhythmias in dogs--effect of celecoxib. AB - A 20-min period of treadmill exercise in dogs, sufficient to increase heart rate by 90-100 beats min(-1), markedly decreases the severity of the life-threatening ventricular arrhythmias that result when, following anaesthesia 24 h later, the left coronary artery is occluded. This protection was unaffected by the administration of the selective cyclooxygenase-2 inhibitor celecoxib (two times 3 mg kg(-1) given intravenously). These results show that prostacyclin, derived from cyclooxygenase-2, plays no role in this pronounced delayed cardioprotection. PMID- 15363968 TI - Ischemic preconditioning of remote organs attenuates gastric ischemia-reperfusion injury through involvement of prostaglandins and sensory nerves. AB - Limitation of the stomach damage by its earlier brief ischemia and reperfusion before prolonged ischemia is defined as gastric ischemic preconditioning but whether such brief ischemia of remote organs like heart or liver can also attenuate the gastric damage caused by longer and severe ischemia-reperfusion remains unknown. The cardiac, hepatic and gastric preconditioning were induced by brief ischemia (occlusion of coronary, hepatic and celiac arteries twice for 5 min) applied 30 min before 3 h of ischemia/reperfusion. Standard 3 h ischemia reperfusion of the stomach produced numerous gastric lesions, decreased gastric blood flow and mucosal prostaglandin E2 generation and increased expression and plasma release of interleukin-1beta and tumor necrosis factor-alpha (TNF-alpha). These effects were significantly attenuated by brief cardiac, hepatic and gastric preconditioning which upregulated cyclooxygenase-2 mRNA but not cyclooxygenase-1 mRNA. The protective effects of brief gastric, cardiac and hepatic preconditioning were attenuated by selective cyclooxygenase-1 and cyclooxygenase 2 inhibitors and capsaicin denervation. We conclude that brief ischemia of remote preconditioning such as heart or liver protects gastric mucosa against severe ischemia-reperfusion-induced gastric lesions as effectively as local preconditioning of the stomach itself via the mechanism involving prostaglandin derived from cyclooxygenase-1 and cyclooxygenase-2 and the activation of sensory nerves releasing calcitonin gene-related peptide (CGRP) combined with the suppression of interleukin-1beta and TNF-alpha expression and release. PMID- 15363969 TI - Current strategies in cancer gene therapy. AB - Cancer gene therapy is the most studied application of gene therapy. Many genetic alterations are involved in the transformation of a normal cell into a neoplastic one. The two main gene groups involved in cancer development are oncogenes and tumor suppressor genes. While the latter eliminates cancerous cells via apoptosis, the former enhances cell proliferation. Therefore, apoptotic genes and anti-oncogenes are widely used in cancer gene therapy. In addition to oncogenes and tumor suppressor genes, chemotherapy and gene therapy can be combined through suicide gene strategy. A suicide gene encodes for a non-mammalian enzyme; this enzyme is used to convert a non-toxic prodrug into its active cytotoxic metabolite within the cancerous cells. Tumor suppressor genes, anti-oncogenes and suicide genes target cancer cells on the molecular level. On the other hand, cancer is immunogenic in nature; therefore, it can also be targeted on the immunological level. Boosting the immune response against cancerous cells is usually achieved via genes encoding for cytokines. Interleukin-12 gene, for example, is one of the most studied cytokine genes for cancer gene therapy applications. DNA vaccines are also used after conventional treatments to eliminate remnant malignant cells. All these therapeutic strategies and other strategies namely anti-angiogenesis and drug resistant genes are briefly reviewed and highlighted in this article. PMID- 15363970 TI - Antiangiogenic versus cytotoxic therapeutic approaches to human pancreas cancer: an experimental study with a vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor and gemcitabine. AB - Pancreatic adenocarcinoma is a leading cause of cancer death in the United States and represents a challenging chemotherapeutic problem. The pharmacological control of angiogenesis might represent a novel approach to the management of pancreas cancer, since the pathological development of vascular supply is a critical step for tumor growth and may affect its prognosis. In order to test this hypothesis, SU5416 ([3-(3,5-dimethyl-1H-pyrrol-2-ylmethylene)-1,3-dihydro indol-2-one]) a selective inhibitor of the vascular endothelial growth factor receptor-2 tyrosine kinase, and gemcitabine (2', 2'-difluorodeoxycytidine) were tested on endothelial (HUVEC) and pancreatic tumor cells (MIA PaCa-2) in vitro and in vivo alone and in simultaneous association. SU5416 inhibited HUVEC cells stimulated to proliferate by vascular endothelial growth factor but not MIA PaCa 2 cells; the drug concentration that decreased cell growth by 50% (IC50) was 0.14 microM. Furthermore, SU5416 reduced the development of microvessels from placental explants (IC50, 0.23 microM). Gemcitabine inhibited the growth of both HUVEC and MIA PaCa-2 cells with an IC50 of 0.08 and 0.1 microM, respectively. A synergistic effect (combination index <1 and dose reduction index >1) on anti proliferative and pro-apoptotic activity was calculated with the simultaneous combination of the two drugs on endothelial cells. A marked in vivo antitumor effect on MIA PaCa-2 xenografts was observed with SU5416 at a protracted schedules, as well as with gemcitabine; furthermore, the combination between the two drugs resulted in an almost complete suppression of tumor growth and relapse. In conclusion, the present results provide the evidence of an effective anti endothelial/antitumor activity of protracted administration of SU5416 on human pancreas cancer xenografts, which is comparable with the one obtained by gemcitabine; moreover, the synergistic combination between these drugs on endothelial cells and the promising association in pancreatic cancer xenografts could be used in future studies and translated into the clinical setting. PMID- 15363971 TI - Inhibition of UVA irradiation-modulated signaling pathways by rutaecarpine, a quinazolinocarboline alkaloid, in human keratinocytes. AB - Matrix metalloproteinases (MMPs), a key component in photoaging of the skin due to exposure to ultraviolet A, appear to be increased by ultraviolet A irradiation associated generation of reactive oxygen species. In this study, we investigated the effects of synthetic rutaecarpine, which is also found in Evodia rutaecarpa, on the ultraviolet A-induced changes in the expression of gelatinases: matrix metalloproteinase (MMP)-2 and MMP-9 using HaCaT human keratinocytes as a model cellular system. Ultraviolet A irradiation of HaCaT cells increased the gelatinolytic activities of MMP-2 and MMP-9, which was significantly suppressed by the pretreatment with rutaecarpine. In addition, rutaecarpine significantly suppressed the ultraviolet A-induced enhanced expression of MMP-2 and MMP-9 proteins and mRNAs. Rutaecarpine also inhibited the H2O2-induced increase in the expression of MMP-2 and MMP-9. Furthermore, rutaecarpine decreased the ultraviolet A-induced increased generation of reactive oxygen species. Taken together, these results suggest that rutaecarpine inhibited ultraviolet A-induced reactive oxygen species generation, resulting in the enhanced expression of MMP-2 and MMP-9 in human skin cells. These results further suggest that ruetaecarpine may be useful in the prevention of ultraviolet A-induced photoaging. PMID- 15363972 TI - Ghrelin and des-acyl ghrelin both inhibit isoproterenol-induced lipolysis in rat adipocytes via a non-type 1a growth hormone secretagogue receptor. AB - Besides possessing a strong growth hormone (GH)-releasing activity, the gastrointestinal octanoylated peptide ghrelin has been reported to antagonize lipolysis in rat adipocytes. It is not yet clear whether this inhibitory activity on lipolysis is also shared by the major circulating isoform, des-acyl ghrelin, that does not activate the ghrelin receptor, namely the type 1a GH secretagogue receptor (GHS-R1a) and lacks the endocrine effects of the acylated form. Here we show that des-acyl ghrelin, like ghrelin and some synthetic GHS (hexarelin and MK0677) and carboxy-terminally ghrelin fragments such as ghrelin-(1-5) and ghrelin-(1-10), all significantly reduced, over concentrations ranging from 1 to 1000 nM, the stimulation of glycerol release caused in rat epididymal adipocytes by the nonselective beta-adrenoceptor agonist isoproterenol in vitro. The order of potency on stimulated-lipolysis was: des-acyl ghrelin=ghrelin>MK0677=hexarelin>ghrelin-(1-5)=ghrelin-(1-10). This ranking was consistent with the binding experiments performed on membranes of epididymal adipose tissue or isolated adipocytes that did not express mRNA for GHS-R1a. A common high-affinity binding site was recognized in these cells by both acylated and des-acylated ghrelin and also by hexarelin, MK0677, ghrelin-(1-5) and ghrelin (1-10). In conclusion, these findings provide the first evidence that des-acyl ghrelin, as well as ghrelin, short ghrelin fragments and synthetic GHS, may act directly as antilipolytic factors on the adipose tissue through binding to a specific receptor which is distinct from GHS-R1a. PMID- 15363973 TI - Sensitization of vanilloid receptor 1 induced by bradykinin via the activation of second messenger signaling cascades in rat primary afferent neurons. AB - Vanilloid receptor 1 was recently reported to play an important role in hyperalgesia, but the mechanisms by which this receptor is activated by endogenous inflammatory mediators, such as bradykinin and nerve growth factor, are not yet fully understood. Here, we investigated whether bradykinin, which is a pain-producing inflammatory mediator, sensitizes vanilloid receptor 1 by inducing the activation of cyclooxygenases, phospholipase C and phospholipase A2 in rat dorsal root ganglion cells. We demonstrated this using 45Ca2+ uptake and inositol phosphates accumulation assays, bradykinin activates phospholipase C and cyclooxygenase-1 through the bradykinin B2 receptor. The bradykinin B2 receptor then sensitizes vanilloid receptor 1 activity by facilitating non-selective Ca2+ channel activity, increasing the intracellular Ca2+ concentration from the extracellular pool. These methods would be useful for screening new drugs for activity at vanilloid receptor 1. These data suggest that endogenous substances produced by several enzymes may be capable of producing a synergistic response involving the vanilloid receptor 1. PMID- 15363974 TI - Andrographolide acts through inhibition of ERK1/2 and Akt phosphorylation to suppress chemotactic migration. AB - We now evaluated the anti-inflammatory mechanisms of andrographolide on complement 5a (C5a)-induced macrophage recruitment in vitro. Andrographolide concentration dependently inhibited cell migration toward C5a with an IC50 of 5.6+/-0.7 microM. With relatively specific kinase inhibitors (PD98059, SB203580, SP600125, wortmannin and LY294002, respectively) the results showed that extracellular signal-regulated kinase1/2 (ERK1/2), p38 mitogen-activated protein kinase (p38 MAPK) and phosphatidylinositol-3-kinase (PI3K) were necessary for C5a induced migration, whereas c-Jun N-terminal kinase (JNK) was nonessential. Andrographolide significantly attenuated C5a-stimulated phosphorylation of ERK1/2, and of its upstream activator, MAP kinase-ERK kinase (MEK1/2). C5a activated ERK1/2 phosphorylation was 86+/-9% inhibited by 30 microM andrographolide. Under the same conditions, however, andrographolide failed to affect C5a-stimulated p38 MAPK and JNK phosphorylation. Andrographolide also strongly abolished C5a-stimulated Akt phosphorylation, a downstream target protein for PI3K. These results indicate that inhibition of cell migration by interfering with ERK1/2 and PI3K/Akt signal pathways may contribute to the anti inflammatory activity of andrographolide. PMID- 15363975 TI - Kappa-opioid receptor stimulation inhibits growth of neonatal rat ventricular myocytes. AB - The effects of trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl]-benzeneacetamide methanesulfonate salt (U50,488H), a selective kappa opioid receptor agonist, on growth in neonatal ventricular myocytes were determined. In 15% serum culture medium, U50,488H at 0.1-1 microM significantly reduced the protein content, [3H]leucine uptake and cell size of the myocytes. The effect of U50,488H on protein content was abolished in the presence of 1 microM nor-binaltorphimine (nor-BNI), a selective kappa-opioid receptor antagonist. In a 0.4% serum medium, U50,488H at 0.1-1 microM had no effect on myocyte growth. Interestingly, 1 microM U50,488H abolished the stimulatory effects of 1 microM norepinephrine on protein content, [3H]leucine uptake and cell size of the myocytes in the low serum medium. The effect of U50,488H was abolished by 1 microM nor-BNI. With the exception of cell size, the effects of norepinephrine were completely abolished by blockade of both alpha- and beta adrenoceptors, but only partially blocked by blockade of either adrenoceptors. These results provide first evidence that kappa-opioid receptor stimulation inhibits growth of the neonatal ventricular myocyte as a result of direct action as well as by inhibiting sympathetic stimulation of the heart. The stimulatory effects of sympathetic activity on growth occurs via both alpha- and beta adrenoceptors. PMID- 15363976 TI - Regulation of serotonin 5-HT2C receptors by chronic ligand exposure. AB - The effect of ligand pretreatment on human 5-hydroxytryptamine2C (5-HT2C) receptors was examined in CHO cells expressing high (CHO-1C7; 67+/-3 pmol/mg) or low (CHO-1C19; 72+/-10 fmol/mg) levels of the receptor. Seventy-two hours pretreatment of CHO-1C7 cells with various ligands did not affect receptor expression. Pretreatment with inverse agonists enhanced 5-HT-mediated inositol phosphate accumulation with no change in constitutive receptor activity. The enhanced agonist responsiveness was inversely correlated with the intrinsic activity of the pretreatment ligand. Seventy-two hours of pretreatment with the weak agonist, 5-methoxygramine, caused an elevation in constitutive activity but no alteration in 5-HT-mediated signaling. In CHO-1C19 cells, 24 but not 72 h of pretreatment with the inverse agonist mianserin enhanced 5-HT-mediated signaling, with no effect on basal signaling; pretreatment with 5-methoxygramine had no significant effect. These findings highlight differences in the pattern of chronic regulation of 5HT2C receptor signaling between high and low receptor expression levels in a common cellular background. PMID- 15363977 TI - Apoptotic activity of betulinic acid derivatives on murine melanoma B16 cell line. AB - The mitochondrion plays a crucial role in the process of apoptosis and has thus become one of the targets for the search for potential chemotherapeutic agents. Betulinic acid [3beta-hydroxy-lup-20(19)lupaen-28-carbonic acid], a lupane-type triterpene which is abundant in many plant species, has been shown to exert a direct effect on the mitochondria and subsequent apoptosis in melanoma cells. Chemical synthesis and modification of betulinic acid are being explored to develop more potent derivatives. We present here the apoptotic activity of several natural derivatives of betulinic acid which were isolated from the roots of a Chinese medicinal herb, Pulsatilla chinensis (Bge) Regel [Ye, W., Ji, N.N., Zhao, S.X., Liu, J.H., Ye, T., McKervey, M.A., Stevenson, P., 1996. Triterpenoids from Pulsatilla chinensis. Phytochemistry 42, 799-802]. Of the five compounds tested, 3-oxo-23-hydroxybetulinic acid was the most cytotoxic on murine melanoma B16 cells (IC50=22.5 microg/ml), followed by 23-hydroxybetulinic acid and betulinic acid (IC50=32 and 76 microg/ml, respectively), with lupeol and betulin exhibiting the weakest cytotoxicity (IC50> or =100 microg/ml). Exposure of B16 cells to betulinic acid, 23-hydroxybetulinic acid and 3-oxo-23-hydroxybetulinic acid caused a rapid increase in reactive oxidative species production and a concomitant dissipation of mitochondrial membrane potential in a dose- and time dependent manner, which resulted in cell apoptosis, as demonstrated by fluorescence microscopy, gel electrophoresis and flow-cytometric analysis. Cell cycle analysis further demonstrated that both 3-oxo-23-hydroxybetulinic acid and 23-hydroxybetulinic acid dramatically increased DNA fragmentation at the expense of G1 cells at doses as low as 12.5 and 25 microg/ml, respectively, thereby showing their potent apoptotic properties. Our results showed that hydroxylation at the C3 position of betulinic acid is likely to enhance the apoptotic activity of betulinic acid derivatives (23-hydroxybetulinic acid and 3-oxo-23 hydroxybetulinic acid) on murine melanoma B16 cells. PMID- 15363978 TI - Inhibition by tranilast of nifedipine-induced proliferation of cultured human gingival fibroblasts. AB - The appropriate method of etiologic therapy for gingival overgrowth is yet unknown. In this study drug-induced proliferation of Gin-1 cells, a normal human gingival fibroblast cell line, was examined by using the reagent water-soluble tetrazolium-1. Tranilast (100 microM) inhibited the nifedipine (10 microM) induced proliferation of gingival fibroblasts. The level of basic fibroblast growth factor (bFGF) was determined by using an enzyme-linked immunosorbent assay kit. Tranilast inhibited the release of bFGF from the cells. In conclusion, tranilast depresses the nifedipine-induced proliferation of gingival fibroblasts by inhibiting the release of bFGF. Administration of tranilast may thus be clinically effective for the treatment of gingival overgrowth. PMID- 15363979 TI - Urantide mimics urotensin-II induced calcium release in cells expressing recombinant UT receptors. AB - Urotensin-II is the natural ligand of the UT receptor. This novel system is involved in the regulation of cardiovascular functions. Recently, a urotensin-II analog ([Pen5,DTrp7,Orn8]urotensin-II(4-11)) named urantide, has been proposed as a selective and potent UT receptor antagonist. In order to pharmacologically characterize this new compound, urantide was tested on the native UT receptors of the rat aorta and on the human recombinant receptors expressed in CHO cells (CHO(hUT)). Indeed, urantide behaves as a competitive, potent (pA2 8.24), and pure antagonist in the rat aorta bioassay, while as an agonist (pEC50 8.11) in a calcium mobilization assay performed in CHO(hUT) cells. Urantide should be considered a low efficacy partial agonist. PMID- 15363980 TI - Opioid agonist and antagonist treatment differentially regulates immunoreactive mu-opioid receptors and dynamin-2 in vivo. AB - Opioid agonists and antagonists can regulate the density of mu-opioid receptors in whole animal and in cell culture. High intrinsic efficacy agonists (e.g., etorphine), but not lower intrinsic efficacy agonists (e.g., morphine), produce mu-opioid receptor down-regulation and can alter the abundance of mu-opioid receptor mRNA. Conversely, opioid antagonists substantially increase the density of mu-opioid receptors without changing its mRNA. Mu-opioid receptor up regulation has been associated with decreases in the trafficking protein dynamin 2, whereas mu-opioid receptor down-regulation produces an increase in dynamin-2 abundance. To probe the differences between opioid agonist and antagonist-induced mu-opioid receptor regulation, the current study determined changes in mu-opioid receptor density using a combined radioligand binding ([3H] DAMGO) and quantitative Western blotting approach in mouse spinal cord. Furthermore, the differences between intermittent and continuous dosing protocols were evaluated. Continuous (7-8 days) s.c. infusions of naloxone (5 mg/kg/day) or naltrexone (15 mg s.c. implant pellet) increased mu-opioid receptor density in radioligand binding assays (approximately +80%) in mouse spinal cord and down-regulated dynamin-2 abundance (approximately -30%), but had no effect on the abundance of immunoreactive mu-opioid receptor. Continuous (7 days) s.c. infusion of etorphine (200 microg/kg/day) decreased immunoreactive mu-opioid receptor (approximately 35%) and [3H] DAMGO binding (approximately -30%), and concurrently increased dynamin-2 abundance (approximately +40%). Continuous (7 days) morphine infusion (40 mg/kg/day plus 25 mg s.c. implant pellet) had no effect on any outcome measure. Delivery of the same daily dose of etorphine or naloxone using intermittent (every 24 h for 7 days) s.c. administration had no effect on immunoreactive mu-opioid receptor, [3H] DAMGO binding or dynamin-2 abundance. These data indicate that mu-opioid receptor density, determined in radioligand binding assays, and immunoreactive dynamin-2 abundance are regulated by continuous, but not intermittent, opioid ligand treatment. Furthermore, the differential regulation of mu-opioid receptor abundance by agonists and antagonists in immunoblotting assays contrasts with changes in [3H] DAMGO binding. Taken together, these results suggest that etorphine-induced down regulation may depend upon mu-opioid receptor degradation and changes in dynamin 2-mediated receptor trafficking. Conversely, antagonist-induced up-regulation does not require an increase in mu-opioid receptor synthesis and may entail conversion of receptors to an appropriate conformation to bind ligand, as well as changes in receptor trafficking. PMID- 15363981 TI - Dopamine D4 receptors inhibit depolarization-induced [3H]GABA release in the rat subthalamic nucleus. AB - We explored the role of dopamine D4 receptors on [3H]GABA release in the subthalamic nucleus. [3H]GABA release was evoked by high K+ in slices of the nucleus. The selective dopamine D4 receptor agonist PD168,077 (N-[[4-(2 cyanophenyl)-1-piperazynil]methyl]-3-methyl-benzamide) inhibited GABA release with greater potency (EC50=3.2 nM) than quinpirole (EC50=200 nM). SKF 21297 (6 chloro-7,8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrobromide), a dopamine D1-like receptor agonist, had no effect. L-745,870 (3-[[4-(4 chlorophenyl)piperazin-1-yl]methyl]-1-1H-pyrollo[2,3-b] pyridine), a selective dopamine D4 receptor antagonist, reverted the quinpirole inhibition with greater potency (IC50=8.7 nM) than that of the dopamine D2/D3 receptor antagonist sulpiride and raclopride (IC50=4804 and 788 nM, respectively). Both methylphenidate and methamphetamine, dopamine reuptake blockers, inhibited by 30% high K(+)-evoked GABA release; the inhibition was blocked by L-745,870. These results show that dopamine D4 receptors modulate GABA release in the subthalamic nucleus. The results would explain how agents that increase interstitial dopamine like methylphenidate and amphethamine might control locomotor hyperactivity seen in disorders of dopamine D4 receptors. PMID- 15363982 TI - Analgesic effect of TT-232, a heptapeptide somatostatin analogue, in acute pain models of the rat and the mouse and in streptozotocin-induced diabetic mechanical allodynia. AB - Somatostatin released from capsaicin-sensitive sensory nerves exerts systemic anti-inflammatory and antinociceptive actions. TT-232 is a stable, peripherally acting heptapeptide (D-Phe-Cys-Tyr-D-Trp-Lys-Cys-Thr-NH2) somatostatin analogue with highest binding affinity for somatostatin sst4 receptors. It has been shown to inhibit acute and chronic inflammatory responses and sensory neuropeptide release from capsaicin-sensitive nociceptors. In the present study the antinociceptive effects of TT-232 were analysed using both acute and chronic models of nociception. Formalin-induced pain behaviour, noxious heat threshold and streptozotocin-induced diabetic neuropathic mechanical allodynia were examined in rats and phenylquinone-evoked abdominal constrictions were tested in mice. TT-232 (80 microg/kg i.p.) inhibited both early (0-5 min) and late phases (25-45 min) of formalin-induced nociception as revealed by determination of the composite pain score. The minimum effective dose to elevate the noxious heat threshold and diminish the heat threshold drop (heat allodynia) evoked by resiniferatoxin (0.05 nmol intraplantarly) was 20 and 10 microg/kg i.p., respectively, as measured by an increasing-temperature hot plate. TT-232 (10-200 microg/kg s.c.) significantly inhibited phenylquinone-evoked writhing movements in mice, but within this dose range no clear dose-response correlation was found. Five weeks after streptozotocin administration (50 mg/kg i.v.) the diabetes induced decrease in the mechanonociceptive threshold was inhibited by 10-100 microg/kg i.p. TT-232. These findings show that TT-232 potently inhibits acute chemical somatic/visceral and thermal nociception and diminishes chronic mechanical allodynia associated with diabetic neuropathy, thereby it could open new perspectives in the treatment of various pain syndromes. PMID- 15363983 TI - Effects of neurosteroids on ischemia-reperfusion injury in the rat retina: role of sigma1 recognition sites. AB - The effects of neurosteroids, 17beta-estradiol and dehydroepiandrosterone-sulfate (DHEA-S), were investigated on retinal degeneration using a rat model of ischemia reperfusion injury. The animals were anaesthetized and retinal ischemia was induced by elevating the intraocular pressure to 120 mm Hg for 45 min. Neurosteroids were injected intraperitoneally before ischemia and immediately after reperfusion. Retinal biochemical changes such as increase of lactate content and decrease of glucose and ATP were significantly inhibited by neurosteroids compared to the control ischemic group. The effects of 17beta estradiol and DHEA-S were antagonized by pre-treatment with the sigma1 site antagonist. These findings suggest that 17beta-estradiol and dehydroepiandrosterone-sulfate may affect the metabolic state of surviving neurons and glial cells after ischemic injury and that they act, at least in part, through involvement of sigma1 recognition sites. PMID- 15363984 TI - Neuroprotective and neurotoxic effects of cyclosporine A on transient focal ischemia in mdr1a knockout mice. AB - The proper dose of cyclosporine A as a neuroprotective agent was investigated using the middle cerebral artery occlusion model of mdr1a knockout mice. After a 30-min occlusion period, reperfusion was performed and the vehicle or cyclosporine A (1 mg/kg or 10 mg/kg x 2) was intraperitoneally administered to each animal model group. Forty eight hours after reperfusion, infarction volume in the 1 mg/kg cyclosporine A group was significantly less than that seen in the vehicle group, although, in the high dose cyclosporine A group, infarction volumes were significantly higher than those seen in the vehicle group. These results demonstrate that cyclosporine A shows not only anti-ischemic effects, but also neurotoxic effects depending on the dosage penetrating into the brain. PMID- 15363985 TI - Suppressing effect of the cannabinoid CB1 receptor antagonist, SR 141716, on alcohol's motivational properties in alcohol-preferring rats. AB - Administration of the cannabinoid CB(1) receptor antagonist, SR 141716 [N piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-3-pyrazole carboxamide], has been reported to reduce alcohol intake and alcohol self administration in different models of excessive alcohol consumption, including the selectively bred Sardinian alcohol-preferring (sP) rats. The present study investigated whether SR 141716 was also capable of decreasing, in this rat line, alcohol's motivational properties. Extinction responding for alcohol, defined as the maximal number of lever responses reached in the absence of alcohol in rats trained to lever-press for alcohol, was used as index of alcohol's motivational properties. Rats were initially trained to lever-press for oral alcohol (15%, v/v) under a fixed ratio (FR) schedule of FR4. Once self-administration behavior was established, extinction sessions were conducted. SR 141716 (0, 0.3, 1 and 3 mg/kg; i.p.) was acutely administered before extinction sessions. In order to assess the specificity of SR 141716 action on extinction responding for alcohol, a separate group of sP rats was trained to lever-press for a 3% (w/v) sucrose solution under an FR4 schedule. SR 141716 administration produced a dose dependent, virtually complete suppression of extinction responding for alcohol. In contrast, extinction responding for sucrose was not significantly altered by treatment with SR 141716. Further to the consummatory aspects, these results also extend the suppressing effect of SR 141716 to the appetitive aspects of alcohol drinking behavior in sP rats. The results also implicate the cannabinoid CB1 receptor in the neural substrate mediating alcohol's motivational properties in this rat line. PMID- 15363986 TI - Within-session repeated acquisition behavior in rats as a potential model of executive function. AB - Higher levels of cognition, such as executive functions, are known to be disrupted in psychiatric disorders such as schizophrenia. As a potential model of executive function, rats were trained in a three-lever operant conditioning chamber to respond on two of the three levers in one of six possible correct sequences. When the rat completed a two-response sequence correctly for 10 consecutive trials, the correct sequence was randomly changed to another two response sequence without signaling the rat. Rats readily acquired the behavioral baseline and completed all six response-sequences within a 60-min session. Phencyclidine, MK-801 ((5S,10R)-(+)-5-Methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine), apomorphine, scopolamine and triazolam all produced dose-related decreases in the total number of sequences completed. Phencyclidine and MK-801 markedly increased all errors while scopolamine produced modest increases; triazolam increased only total and intrarule errors, while apomorphine had no significant effect on errors. The present results suggest that within-session repeated acquisition of response sequences has the potential to be a useful model for studying executive function in rats. PMID- 15363987 TI - Effects of rolipram, a phosphodiesterase 4 inhibitor, in combination with imipramine on depressive behavior, CRE-binding activity and BDNF level in learned helplessness rats. AB - The brain cAMP regulating system and its downstream elements play a pivotal role in the therapeutic effects of antidepressants. We previously reported the increase in activities of phosphodiesterase 4, a major phosphodiesterase isozyme hydrolyzing cAMP, in the frontal cortex and hippocampus of learned helplessness rats, an animal model for depression. The present study was undertaken to examine the combination of effects of rolipram, a phosphodiesterase 4 inhibitor, with imipramine, a typical tricyclic antidepressant, on depressive behavior in learned helplessness rats. Concurrently, cAMP-response element (CRE)-binding activity and brain-derived neurotrophic factor (BDNF) levels related to the therapeutic effects of antidepressants were determined. Repeated administration of imipramine (1.25-10 mg/kg, i.p.) or rolipram (1.25 mg/kg, i.p.) reduced the number of escape failures in learned helplessness rats. Imipramine could not completely ameliorate the escape behavior to a level similar to that of non-stressed rats even at 10 mg/kg. However, repeated coadministration of rolipram with imipramine (1.25 and 2.5 mg/kg, respectively) almost completely eliminated the escape failures in learned helplessness rats. The reduction of CRE-binding activities and BDNF levels in the frontal cortex or hippocampus in learned helplessness rats were ameliorated by treatment with imipramine or rolipram alone. CRE-binding activities and/or BDNF levels of the frontal cortex and hippocampus were significantly increased by treatment with a combination of rolipram and imipramine compared to those in imipramine-treated rats. These results indicated that coadministration of phosphodiesterase type 4 inhibitors with antidepressants may be more effective for depression therapy and suggest that elevation of the cAMP signal transduction pathway is involved in the antidepressive effects. PMID- 15363988 TI - Opioid and monoamine systems mediate the discriminative stimulus of tramadol in rats. AB - We analyzed the ability of the mu opioid peptide receptor ligands morphine and naloxone and several antidepressant drugs that are serotonin (fluoxetine), noradrenaline (reboxetine), mixed serotonin and noradrenaline (milnacipram and venlafaxine), dopamine (nomifensine) reuptake inhibitors, as well as roxindole (a nonselective drug) to substitute for, or alter, tramadol discrimination. Male Wistar rats were trained to discriminate tramadol (20 mg/kg) from saline in a two choice water-reinforced paradigm. Out of the drugs studied, only morphine substituted for tramadol. In combination experiments, naloxone (0.1-1 mg/kg) attenuated the stimulus effects of tramadol (20 mg/kg) and the substitution evoked by morphine (2 mg/kg). Milnacipram (10 mg/kg) or reboxetine (10 mg/kg) enhanced the effects of tramadol (2.5-10 mg/kg); the other antidepressant drugs used failed to modulate tramadol discrimination. Our results indicate that tramadol can be used as a stimulus cue in rats, and that mu opioid peptide mechanisms are involved in its effects, while noradrenergic uptake inhibitors can enhance tramadol discrimination. PMID- 15363989 TI - Anxiolytic-like effects of PHCCC, an allosteric modulator of mGlu4 receptors, in rats. AB - We examined the potential anxiolytic-like activity of (-)-N-phenyl-7 (hydroxyimino) cyclopropa[b]chromen-1a-carboxamide (PHCCC), an allosteric modulator of metabotropic glutamate4 receptors (mGlu4), after administration into the basolateral amygdala, using the conflict drinking Vogel test in rats as a model. The results indicate that PHCCC, but not 7 (hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOEt), the selective antagonist of group mGlu1 receptors, showed significant, dose-dependent anticonflict effects without affecting the threshold current or water intake. The results indicate that positive allosteric modulation of mGlu4 receptors may be a useful therapeutic approach to anxiety. PMID- 15363990 TI - Behavioral sensitization to the discriminative stimulus effects of methamphetamine in rats. AB - Sensitization to the discriminative stimulus effects of psychostimulants is not fully understood. Therefore, the present study was designed to investigate the development of sensitization to the discriminative stimulus of methamphetamine in rats. A dose-response curve for methamphetamine and a generalization test for cocaine were recorded in rats trained to discriminate between 1.0 mg/kg methamphetamine and saline. A significant leftward shift of the dose-response curve for methamphetamine and of the dose-generalization curve for cocaine was observed following repeated administration of methamphetamine (2.0 mg/kg) instead of saline. These findings suggest that repeated administration of methamphetamine can produce behavioral sensitization to the discriminative stimulus effects of methamphetamine in rats. PMID- 15363991 TI - Lys-[Leu8,des-Arg9]-bradykinin blocks lipopolysaccharide-induced SHR aorta hyperpolarization by inhibition of Ca(++)- and ATP-dependent K+ channels. AB - The mediators involved in the hyperpolarizing effects of lipopolysaccharide and of the bradykinin B1 receptor agonist des-Arg9-bradykinin on the rat aorta were investigated by comparing the responses of aortic rings of spontaneously hypertensive and normotensive Wistar rats. Endothelized rings from hypertensive rats were hyperpolarized by des-Arg9-bradykinin and lipopolysaccharide, whereas de-endothelized rings responded to lipopolysaccharide but not to des-Arg9 bradykinin. In endothelized preparations, the responses to des-Arg9-bradykinin were inhibited by Nomega-nitro-L-arginine and iberiotoxin. De-endothelized ring responses to lipopolysaccharide were inhibited by iberiotoxin, glibenclamide and B1 antagonist Lys-[Leu8,des-Arg9]-bradykinin. This antagonist also inhibited hyperpolarization by des-Arg9-bradykinin and by the a2-adrenoceptor agonist, brimonidine. Our results indicate that Ca(2+)-sensitive K+ channels are the final mediators of the responses to des-Arg9-bradykinin, whereas both Ca(2+)- and ATP sensitive K+ channels mediate the responses to lipopolysaccharide. The inhibitory effects of Lys-[Leu8,des-Arg9]-bradykinin is due to a direct action on Ca(2+)- and ATP-sensitive potassium channels. PMID- 15363992 TI - Superiority of YM598 over atrasentan as a selective endothelin ETA receptor antagonist. AB - The binding affinities of (E)-N-[6-methoxy-5-(2-methoxyphenoxy)[2,2'-bipyrimidin] 4-yl]-2-phenylethenesulfonamide monopotassium salt (YM598) for native human endothelin ETA and ETB receptors expressed in human coronary artery smooth muscle cells (CASMC) and a human melanoma cell line, SK-Mel-28, respectively, were examined, and the results compared with those for the endothelin receptor antagonists atrasentan and bosentan. The in vivo endothelin ETA receptor inhibitory activities of YM598 and atrasentan were also compared through the suppression of the big endothelin-1-induced pressor response in pithed rats. Ki values of YM598, atrasentan, and bosentan for native human endothelin ETA receptors were 0.772, 0.0551, and 4.75 nM, while those for native human endothelin ETB receptors were 143, 4.80, and 40.9 nM, respectively. The calculated selectivity ratios of YM598, atrasentan, and bosentan for endothelin ETA versus ETB receptors were 185, 87 and 8.6, respectively. In pithed rats, YM598 and atrasentan inhibited the big endothelin-1 (1 nmol/kg)-induced pressor response in a dose-dependent manner on both intravenous and oral administration. The inhibitory effect of YM598 was less potent than that of atrasentan when these agents were intravenously administered, but closely similar on oral administration. These results suggest that YM598 has high selectivity for native human ETA against ETB receptors, and that YM598 is superior to atrasentan as an ETA receptor antagonist with regard to pharmacological bioavailability in rats. PMID- 15363993 TI - Restoration of middle cerebral artery thrombosis by novel glycoprotein IIb/IIIa antagonist FK419 in guinea pig. AB - We compared the antithrombotic efficacy of FK419 [(S)-2-acetylamino-3-[(R)-[1-[3 (piperidin-4-yl)propionyl]piperidin-3-ylcarbonyl]amino] propionic acid trihydrate], a novel nonpeptide glycoprotein IIb/IIIa antagonist, with recombinant tissue plasminogen activator (rt-PA) and other antithrombotic agents (aspirin, ozagrel, argatroban and heparin). FK419 not only inhibited ADP- and collagen-induced guinea pig platelet aggregation, but also induced disaggregation for ADP-induced aggregated platelets in vitro. In the photochemically induced middle cerebral artery thrombosis model in guinea pigs, FK419 dose-dependently shortened the time to first reperfusion and the total middle cerebral artery occlusion time and reduced ischemic brain damage and ameliorated neurological deficits measured 24 h after middle cerebral artery occlusion. Rt-PA similarly improved the middle cerebral artery patency, brain damage and neurological deficits. Neither aspirin, ozagrel, argatroban nor heparin restored the middle cerebral artery blood flow and improved the brain damage or neurological deficits. These results demonstrated that novel glycoprotein IIb/IIIa antagonist FK419 could disperse thrombus and ameliorated ischemic brain damage, suggesting that FK419 would be an attractive intervention for stroke patients. PMID- 15363994 TI - Atorvastatin enhances sildenafil-induced vasodilation through nitric oxide mediated mechanisms. AB - Statins have cholesterol-independent effects including an increased vascular nitric oxide (NO) activity and are commonly used by patients with cardiovascular disease. Such patients frequently have erectile dysfunction, which may be treated with sildenafil, a selective inhibitor of phosphodiesterase type 5. Since statins and sildenafil can activate the NO-cGMP pathway, we investigated whether pre treatment with atorvastatin (0, 5 and 30 mg/kg/day) for 2 weeks affects sildenafil (1 pM-100 mM)-induced relaxation of aortic rings isolated from Wistar rats. We also examined the hemodynamic consequences of this interaction in Wistar rats. Plasma nitrite/nitrate (NOx) concentrations were determined using an ozone based chemiluminescence assay. While pre-treatment with atorvastatin increased the potency of sildenafil-induced vasorelaxation (P<0.01), no differences were observed in the maximum sildenafil-induced relaxation. Pre-incubation of aortic rings with NG-nitro-L-arginine methyl ester (L-NAME) reversed atorvastatin induced increase in the potency of sildenafil relaxation. In addition, pre treatment with atorvastatin enhanced plasma NOx concentrations and sildenafil induced hypotension and tachycardia (all P<0.05). These results suggest that atorvastatin increases the vascular sensitivity to sildenafil through NO-mediated mechanisms. PMID- 15363995 TI - Synergistic interaction of endogenous platelet-activating factor and vasopressin in generating angina in rats. AB - We examined the involvement of endogenous vasopressin and platelet-activating factor (PAF) in the pathogenesis of two types of experimental angina in urethane anaesthetised male Wistar rats. In the first model, epinephrine (10 microg kg( 1)) was injected into the tail vein, followed at the development of the maximum blood pressure response, i.e., 30 s later, by phentolamine (15 mg kg(-1)). In the second model, the vasopressin V1 receptor agonist ornithine-vasopressin (ornipressin; 0.5 IU kg(-1), i.v.) was administered. The heart rate, mean arterial blood pressure and surface electrocardiogram (ECG, standard lead II) were registered simultaneously. As a measure of myocardial ischaemia, at 1 min after phentolamine or ornipressin administration, we found significant ST-segment depression, lasting for more than 10 or 5 min, respectively. Pretreatment (15 min, s.c.) with the vasopressin V1 receptor antagonist Mca1,Tyr(Me)2AVP (the Manning peptide; 0.02-0.2 microg kg(-1)) or the platelet-activating factor receptor antagonist ginkgolide B (BN 52021; 0.25-2.5 mg kg(-1)) alone caused a dose-dependent reduction of the ST-segment depression. Concurrent administration of the two antagonists in their threshold doses (0.02 microg kg(-1) and 0.25 mg kg(-1)) also attenuated the ST-segment depression in both models. Neither antagonist affected the blood pressure or heart rate changes throughout the studies. Our results suggest that endogenous vasopressin and platelet-activating factor interact synergistically in provoking myocardial ischaemia in vivo in experimental angina in the rat. PMID- 15363996 TI - Differences in the vascular selectivity and tolerance between the NO donor/beta blocker PF9404C and nitroglycerin. AB - The vascular selectivity of PF9404C ((2'S),(2S)-3-isopropylamine,1-[4-(2,3 dinitroxy)propoxymethyl]-phenoxy-2'-propanol), a new beta-blocker with nitric oxide (NO)-donor and vasodilator properties, was studied in different rabbit arteries and veins. Phenylephrine (10(-6) M) or 35 mM K+ were used to pre contract the arteries and veins prior to study the relaxant effects of PF9404C and nitroglycerin. The potency of both drugs to depress the phenylephrine-induced contraction was greater than that shown in the blockade of the K(+)-evoked contraction in most of the vessels studied, with the exception of the central ear artery. PF9404C exhibited about three-fold higher potency than nitroglycerin to relax the majority of the vessels studied, especially when they were contracted with K+, and showed a certain selectivity of action for the renal artery. PF9404C produced autotolerance but this effect was about 20-fold less pronounced than that observed with nitroglycerin. Cross-tolerance in those preparations pre exposed to PF9404C that were relaxed later on with nitroglycerin was much greater than autotolerance. The tolerance for nitroglycerin was practically abolished in the presence of N-acetylcysteine. However, this was not the case for PF9404C. These results indicate that, although sharing the property of being NO donors, PF9404C and nitroglycerin show a different profile in causing vasodilation; furthermore, the tolerance to this effect is lesser for PF9404C and seems to be mediated by a mechanism different to that of nitrates. This makes PF9404C a nice pharmacological tool to further develop novel NO-donor compounds with a lesser degree of vascular tolerance than those now available. PMID- 15363997 TI - Upregulation of Rho-kinase (ROCK-2) expression and enhanced contraction to endothelin-1 in the mesenteric artery from lipopolysaccharide-treated rats. AB - Effects of bacterial lipopolysaccharide (Escherichia coli serotype, 055:B5, 20 mg kg(-1), i.p., for 6 h) and a Rho-kinase inhibitor, (+)-(R)-trans-4-(1-aminoethyl) N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride monohydrate, Y-27632 (10(-9) 10(-5) M) were investigated on the contractile responses of the rat mesenteric artery to phenylephrine (10(-9)-3 x 10(-5) M), angiotensin-2 (10(-10)-10(-6) M) and endothelin-1 (10(-10)-10(-7) M). Moreover, alteration in the level of Rho kinase (ROCK-2) expression was examined in the superior mesenteric artery obtained from saline- and lipopolysaccharide-treated rats by Western blotting. Endotoxemic rat mesenteric rings exhibited no different contractions to phenylephrine and angiotensin-2 but augmented contractile activity to endothelin 1. In the mesenteric artery obtained from the endotoxemic rats, acetylcholine induced vasorelaxation did not differ; pD2 value for acetylcholine was 7.85+/ 0.12 in the endotoxemic rings; however, it was 7.81+/-0.15 in the control rings (P>0.05). Y-27632 induced relaxation, which was the same in the control arteries as in endotoxemic ones when contracting agent was phenylephrine. However, when endothelin-1 was used to precontract the rings, Y-27632 produced enhanced relaxation in endotoxemic vessels. pD2 values for Y-27632 were, respectively, 7.69+/-0.12 and 8.20+/-0.10 in control and endotoxemic rings precontracted by endothelin-1 (10(-8) M) (P<0.01). Moreover, Y-27632 (10(-5) M) suppressed the contraction induced by angiotensin-2 (10(-10)-10(-6) M). Western blot analysis revealed that Rho-kinase was upregulated significantly in the mesenteric artery obtained from the rats treated with LPS for 6 h. In addition, serum NO2-/NO3- level, which was detected by Griess method, was 10.0+/-1.4 microM in endotoxemic rats; however, it was 6.6+/-0.5 microM in control (P<0.05). Taken together, these results show that the expression of the contractile protein Rho-kinase could be upregulated in endotoxemic mesenteric artery and this upregulation may be coincided with an enhanced contraction to endothelin-1 but not phenylephrine and angiotensin-2. PMID- 15363998 TI - Pharmacological characterisation of the thermogenic effect of bupropion. AB - The pharmacological mechanism of bupropion's thermogenic effect has been investigated in female Wistar rats by measuring oxygen consumption at thermoneutrality (29 degrees C). Bupropion (30 mg/kg) rapidly increased oxygen consumption (VO2) with a maximum effect at 30 min, and VO2 remained elevated throughout the 4-h experimental period. The nonselective 5-hydroxytryptamine (5 HT or serotonin) receptor antagonist, metergoline (1 mg/kg), and the alpha1 adrenoceptor antagonist, prazosin (1 mg/kg), had no effect on the VO2 response to bupropion, whereas the alpha2-adrenoceptor antagonist, RS79948 [(8aR, 12aS, 13aS) 5,8,8a,9,10,11,12,12a,13,13a-decahydro-3-methoxy-12-(ethylsulphonyl)-6H isoquino[2,1-g][1,6]-naphthyridine hydrochloride] (1 mg/kg), potentiated the response. The VO2 response to bupropion during the first 60 min was significantly inhibited by a high dose of the nonselective beta-adrenoceptor antagonist, propranolol (20 mg/kg), but it had no effect at a low dose (1 mg/kg). Pretreatment with the dopamine D2/D1 receptor antagonist, (+)butaclamol (200 microg/kg), caused a partial, but significant, inhibition (P<0.01) of the VO2 response to bupropion during the first 60 min, and this antagonist abolished the effect of bupropion between 90 and 240 min. Pretreatment with a combination of a high dose of propranolol (20 mg/kg) and (+)butaclamol (200 microg/kg) prevented any increase in VO2 induced by bupropion. It is concluded that the beta3 adrenoceptor subtype, as well as dopamine D2/D1 receptors, is responsible for the increase in oxygen consumption induced by bupropion. We have previously demonstrated that bupropion did not significantly reduce food intake in rats. Hence, in this species, its weight-reducing action predominantly results from thermogenesis mediated via activation of beta3-adrenergic and dopamine D2/D1 receptors. Because bupropion has also been reported not to alter food intake in the clinic, thermogenesis may also contribute to its antiobesity effect in man. PMID- 15363999 TI - Interaction between adenosine and allergen or compound 48/80 on lung parenchymal strips from actively sensitized Brown Norway rats. AB - We have investigated the effect of mast cell activation induced by immunological and non-immunological stimuli on the sensitivity to adenosine of parenchymal strips prepared from lungs removed from Brown Norway (BN) rats actively sensitized to ovalbumin. Strips responded to ovalbumin with a biphasic contractile response. Responses to adenosine were markedly increased 30 min after ovalbumin. The first phase of the response to ovalbumin was abolished by the 5 hydroxytryptamine (5-HT)2 receptor antagonist, methysergide and unaffected by the cysteinyl leukotriene receptor antagonist, iralukast. The second phase was abolished by iralukast and unaffected by methysergide. The response to adenosine was markedly reduced by methysergide but not significantly altered by iralukast. Compound 48/80 (condensation product of N-methyl-p-methoxyphenylethylamine with formaldehyde) induced methysergide-sensitive contractions of the parenchymal strip and potentiated adenosine; the augmented response to adenosine was blocked by methysergide. Thus, activation of mast cells in the lung by either immunological or non-immunological stimuli results in augmentation of the mast cell-mediated contractile response to adenosine. PMID- 15364000 TI - In vivo and in vitro study of the influence of the anticholinesterase drug galantamine on motor and evacuative functions of rat gastrointestinal tract. AB - Galantamine is efficacious for vascular dementia and Alzheimer's disease. Its application leads to some negative gastrointestinal side effects. The present study observes galantamine-induced influence on gastrointestinal motility of rats and its effects on isolated gastrointestinal smooth muscles. The gastrointestinal tract was studied by X-ray contrast examination. Functional disturbances were observed: hypertonia, increased stomach and ileal peristalsis activity, accelerated intestinal passage. In vitro, the drug caused tonic contractions in smooth muscle preparations and increased the gastric and ileal phasic amplitude. The jejunal smooth muscle strips demonstrated an opposite tendency. The reactions were a result of the interaction of galantamine-accumulated endogenic acetycholine with M- and N-acetylcholine receptors. The tonic effects were influenced in varying degree by atropine and ipratropium, whereas the phasic by atropine, ipratropium, hexametonium and methysergide. In conclusion, the in vitro effects registered satisfactorily explain in vivo examined galantamine-induced changes in the gastrointestinal tract of the treated rats and can be considered as main cause for development of such changes. PMID- 15364001 TI - Mechanisms underlying postjunctional synergism between responses of the vas deferens to noradrenaline and ATP. AB - Mechanisms of postjunctional synergism between adenosine 5'-triphosphate (ATP) and noradrenaline were studied in isolated guinea pig vas deferens. Whereas prior exposure to ATP had no significant effect on noradrenaline-mediated contractions, noradrenaline concentration-dependently enhanced ATP-induced contractions. Similarly to noradrenaline, histamine, which also acts via phospholipase-coupled receptors, induced contractions of the vas deferens and enhanced subsequent responses to ATP. Although phorbol-12, 13-dibutyrate (PDBu), a stimulant of protein kinase C (PKC), failed to induce contractions, it significantly potentiated ATP-induced contractions. The PKC inhibitor, Calphostin C, prevented this effect and the noradrenaline-mediated enhancement of ATP-induced contractions. The phosphatase inhibitor cantharidin induced a time- and concentration-dependent tonic contraction and markedly increased subsequent contractions to ATP. It is suggested that noradrenaline potentiates the contractile response of the vas deferens to ATP via a PKC-mediated mechanism. This may involve the inhibition of myosin light chain phosphatase (MLCP) and subsequent calcium sensitisation. PMID- 15364002 TI - Opioid profiles of Cys2-containing enkephalin analogues. AB - To elucidate the structural features determining delta-opioid receptor properties of enkephalin analogues containing Cys(O2NH2) in position 2, a series of Cys2 containing derivatives were synthesized and tested for their effectiveness in depressing electrically evoked contractions of the mouse vas deferens (predominantly enkephalin-selective delta-opioid receptors) and the guinea-pig ileum (mu- and kappa-opioid receptors). The peptidase resistance of the compounds was also tested. The ratio IC50 in the guinea-pig ileum/IC50 in the mouse vas deferens, indicating selectivity for delta-opioid receptors, was high for Cys(O2NH2)2-containing analogues and especially for [Cys(O2NH2)2, Leu5]enkephalin, which was about seven times more selective than delta-opioid receptor selective ligand cyclic [D-Pen2, D-Pen5]enkephalin (DPDPE). The dissociation constant (KA) and relative efficacy (e(rel)) of the compounds in the mouse-isolated vas deferens were determined using explicit formulae derived by fitting of the data points with two-parametric hyperbolic function. The obtained values for KA and e(rel) suggest that: (i) incorporation of Cys(O2NH2)2 in the molecule of [Leu5]enkephalin highly increases the efficacy and does not change significantly the affinity of the respective analogues to delta-opioid receptors; [Cys(O2NH2)2, Leu5]enkephalin has higher affinity than DPDPE, but is less resistant to enzyme degradation; the effect of this modification on the efficacy is decreased when methionine is in position 5; (ii) D-configuration of Cys(O2NH2)2-containing analogues increases their peptidase resistance, but reduces efficacy and affinity of the peptides towards delta-opioid receptors; (iii) the substitution of Cys(O2NH2) with Hcy(O2NH2) reduces the efficacy, affinity and potency of the respective analogues and maintains their sensitivity to endogenous peptidases; (iv) the substitution of the sulfonamide group with benzyl group in the molecule of Cys in position 2 decreases their efficacy and affinity toward delta-opioid receptors, but attaches resistance to enzyme degradation. The results obtained in this study allow: (i) to involve the receptor affinity and agonist efficacy as drug-design consideration for delta opioid receptor properties of newly synthesized compounds and (ii) to characterize some of the structural features, which set the pattern for their opioid profiles. PMID- 15364003 TI - Effect of pioglitazone on endotoxin-induced decreases in hepatic drug metabolizing enzyme activity and expression of CYP3A2 and CYP2C11. AB - It has been reported that peroxisome proliferator-activated receptor-gamma (PPAR gamma) ligands ameliorate the expression of inducible nitric oxide synthase (iNOS) by endotoxin. In the present study, we investigated the effect of pioglitazone, a potent PPAR-gamma ligand, on the endotoxin-induced reduction of hepatic drug-metabolizing enzyme activity and on the down-regulation of the expression of hepatic cytochrome P450 (CYP) 3A2 and CYP2C11 proteins in rats. Endotoxin (1 mg/kg) significantly decreased hepatic drug-metabolizing enzyme activity in vivo, as represented by the systemic clearance of antipyrine and protein levels of CYP3A2 and CYP2C11 24 h after intraperitoneal injection. Pretreatment with pioglitazone (10 mg/kg, 4 times at 10-min intervals) significantly protected the endotoxin-induced decreases in the systemic clearance of antipyrine and protein levels of CYP3A2, but not CYP2C11, with no biochemical and histopathological changes in the liver. Pioglitazone alone had no effect on the systemic clearance of antipyrine and protein levels of CYP3A2 or CYP2C11. Pioglitazone significantly protected endotoxin-induced overexpression of iNOS in the liver, but not the overproduction of nitric oxide (NO) in plasma. It is unlikely that the protective effect of pioglitazone against endotoxin-induced decreases in the hepatic drug-metabolizing enzyme activity and protein levels of CYP3A2 in the liver is due to the inhibition of the overproduction of NO. PMID- 15364004 TI - Blood flow-dependent changes in intrarenal nitric oxide levels during anesthesia with halothane or sevoflurane. AB - We previously demonstrated that intrarenal nitric oxide (NO) levels and renal blood flow are reduced during halothane anesthesia. Studies were performed to determine if volatile anesthetics-induced reductions in renal NO levels are associated with blood flow changes. Halothane and sevoflurane at 0.8 and 2.4 Mac were administered by inhalation to dogs, and cGMP and NOx concentrations in the renal interstitial fluid were measured by a microdialysis method. Neither halothane nor sevoflurane at 0.8 Mac altered renal blood flow and renal interstitial cyclic guanosine monophosphate (cGMP) and NOx levels, but both anesthetics significantly decreased these values at 2.4 Mac. Using an adjustable aortic clamp, renal perfusion pressure was reduced in 2 steps without halothane and sevoflurane anesthesia. Renal blood flow as well as cGMP and NOx concentrations in the renal interstitial fluid were unchanged within the autoregulatory range, but significantly decreased below the autoregulatory range. Changes in cGMP and NOx concentrations in the renal interstitial fluid were highly correlated with renal blood flow changes during halothane or sevoflurane anesthesia, and during stepwise reductions in renal perfusion pressure. The results suggested that halothane- and sevoflurane-induced decreases in intrarenal NO levels result from reductions in blood flow. PMID- 15364005 TI - Inhibition of cyclooxygenase-2, but not cyclooxygenase-1, reduces prostaglandin E2 secretion from diabetic rat retinas. AB - Up-regulation of cyclooxygenase-2 occurs in retinal cells during the early onset of diabetic retinopathy. Under these conditions, prostaglandin production is elevated, which in turn leads to an increased expression of vascular endothelial growth factor (VEGF)--a growth factor implicated in vascular leakage and neovascularization. In this ex vivo study, we tested whether cyclooxygenase-1 or cyclooxygenase-2 is responsible for diabetes-induced secretion of prostaglandin E2 from isolated rat retinas. Celecoxib, a selective cyclooxygenase-2 inhibitor, significantly inhibited prostaglandin E2 secretion, whereas SC560 [5-(4 chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethylpyrazole], a selective cyclooxygenase-1 inhibitor, had no inhibitory effect. These results suggests that the enzymatic activity of cyclooxygenase-2, but not cyclooxygenase-1, results in prostaglandin E2 secretion under diabetic conditions. PMID- 15364006 TI - Inhibition of interleukin-1beta reduces mouse lung inflammation induced by exposure to cigarette smoke. AB - We examined nuclear factor kappaB activation, release of inflammatory mediators and cellular infiltration in acute cigarette smoke inflammation models. One hour after exposure to one puff of cigarette smoke, alveolar macrophages from bronchoalveolar lavage (BAL) fluid of C57BL/6J mice showed an increased activity of nuclear factor kappaB-DNA binding but similar numbers as compared to that of BAL fluid from mice exposed to ambient air. Exposure to 1 cigarette/day for 1, 4 or 7 days led to an increase in interleukin-1beta and monocyte chemoattractant protein-1 levels and to a progressive influx of nuclear factor kappaB-activated alveolar macrophages into the BAL fluid and lung tissue. Exposure to 2 cigarettes/day for 7 days led to a significant increase in interleukin-1beta levels accompanied by a massive alveolar macrophage influx into the BAL fluid. Tumor necrosis factor-alpha levels and subsequent neutrophil influx were only detected after exposure to 4 or 8 cigarettes/day for 7 days. Treatment of mice with an antibody anti-interleukin-1beta during cigarette smoke exposure for 7 days significantly reduced both interleukin-1beta levels and alveolar macrophage influx. These data show that a single exposure to cigarette smoke rapidly activates alveolar macrophages, inducing the production of interleukin-1beta, which may play an important role in triggering chronic cigarette smoke-mediated lung inflammation. PMID- 15364007 TI - Involvement of enhanced neurokinin NK3 receptor expression in the severe asthma guinea pig model. AB - In this study, we investigated the involvement of neurokinin NK3 receptors in a severe asthma model prepared by administering ovalbumin via inhalation three times to systemically sensitized guinea pigs. [3H]senktide, a neurokinin NK3 receptor ligand, showed significant specific binding to the lungs from the model animals, but not to those from negative control animals. The airway responsiveness to intravenous neurokinin B, a neurokinin NK3 receptor agonist, was increased in the model, indicating an increase in functional NK3 receptors. Furthermore, SB 223956 ((-)-3-methoxy-2-phenyl-N-[(1S)-phenylpropyl]quinoline-4 carboxamide), a selective neurokinin NK3 receptor antagonist, significantly inhibited the ovalbumin-induced airway hyperresponsiveness to inhaled methacholine, but it did not show significant effects on the ovalbumin-induced airway narrowing and eosinophil accumulation. These results suggest that the expressed neurokinin NK3 receptors in the severe asthma model are involved in the development of airway hyperresponsiveness. PMID- 15364008 TI - 15d-prostaglandin J2 reduces multiple organ failure caused by wall-fragment of Gram-positive and Gram-negative bacteria. AB - Septic shock is still the major cause of death in surgical intensive care units. Both gram-positive (G+) and gram-negative (G-) bacteria have been isolated in the blood of a large portion of septic patients, and these polymicrobial infections often have a higher mortality than infections due to a single organism. Cell wall fragments from G+ and G- bacteria synergise to cause shock and multiple organ dysfunction in vivo (G+/G- shock). Male Wistar rats were anaesthetised and received a coadministration of wall fragments from G+ and G- bacteria, Staphilococcus aureus (S. aureus) peptidoglycan [0.3 mg/kg, intravenously (i.v.)] and Escherichia coli (E. coli) lipopolysaccharide (1 mg/kg, i.v.) or vehicle (saline, 1 ml/kg, i.v.). G+/G- shock for 6 h resulted in an increase in serum levels of creatinine (indicator of renal dysfunction), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (gamma-GT), bilirubin (markers for hepatic injury and dysfunction) and creatine kinase (CK, an indicator of neuromuscular, skeletal muscle or cardiac injury). Pretreatment of rats with the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist 15d-prostaglandin J2 (0.3 mg/kg, i.v., 30 min prior to G+/G-) reduced the multiple organ injury/dysfunction caused by coadministration of peptidoglycan+lipopolysaccharide. The selective PPAR-gamma antagonist GW9662 (2 Chloro-5-nitrobenzanilide) (1 mg/kg, i.v., given 45 min prior to G+/G-) abolished the protective effects of 15d-prostaglandin J2. 15d- prostaglandin J2 did not affect the biphasic fall in blood pressure or the increase in heart rate caused by administration of peptidoglycan+lipopolysaccharide. The mechanism(s) of the protective effect of this cyclopentenone prostaglandin are-at least in part-PPAR gamma dependent, as the protection afforded by 15d-prostaglandin J2 was reduced by the PPAR-gamma antagonist GW9662. We propose that 15d-prostaglandin J2 or other ligands for PPAR-gamma may be useful in the therapy of the organ injury associated with septic shock. PMID- 15364009 TI - Effects of benidipine, a dihydropyridine-Ca2+ channel blocker, on expression of cytokine-induced adhesion molecules and chemoattractants in human aortic endothelial cells. AB - Benidipine hydrochloride (benidipine) is a dihydropyridine-Ca2+ channel blocker with antioxidant properties. We examined the effects of benidipine on cytokine induced expression of adhesion molecules and chemokines, which play important roles in the adhesion of monocytes to endothelium. Pretreatment of human aortic endothelial cells (HAECs) with benidipine (0.3-10 micromol/l) for 24 h significantly suppressed cytokine-induced vascular cell adhesion molecule-1 (VCAM 1) and intracellular cell adhesion molecule-1 (ICAM-1) mRNA and protein expression, resulting in reduced adhesion of THP-1 monocytes. Benidipine also suppressed induction of monocyte chemoattractant protein (MCP)-1 and interleukin 8. Benidipine inhibited redox-sensitive transcriptional nuclear factor-kappaB (NF kappaB) pathway, as determined by Western blotting of inhibitory kappaB (IkappaB) phosphorylation and luciferase reporter assay. Results of analysis using optical isomers of benidipine and antioxidants suggested that these inhibitory effects were dependent on pharmacological effects other than Ca2+ antagonism such as antioxidant effects. Benidipine may thus have anti-inflammatory properties and benefits for in the treatment of atherosclerosis. PMID- 15364010 TI - Protective role of NF-kappaB1 (p50) in experimental pneumococcal meningitis. AB - Nuclear factor-kappaB (NF-kappaB) is a critical regulator of many genes involved in the pathogenesis of bacterial meningitis. Recently, activation of NF-kappaB was shown to be a key event in the inflammatory host response and the development of intracranial complications during experimental pneumococcal meningitis. Since the p50 subunit of NF-kappaB lacks a transactivation domain and can therefore act as a transcriptional repressor, we investigated whether NF-kappaB1 (p50) exerts anti-inflammatory effects in pneumococcal meningitis. p50-deficient mice had higher cerebellar pneumococcal titers (10.06+/-0.47 vs. 8.51+/-1.06 log colony forming units [cfu]/cerebellum), cerebrospinal fluid (CSF) leukocyte counts (11,475+/-2340 vs. 8444+/-1405 cells/microl) and brain concentrations of interleukin-1beta (125.9+/-50.3 vs. 58.5+/-52.2 pg/mg protein) than their wild type littermates. With ceftriaxone therapy, none of the wild-type mice but 43% of the p50-deficient animals died. In conclusion, lack of NF-kappaB1 (p50) was associated with impaired bacterial clearing, enhanced inflammatory host response and increased mortality during pneumococcal meningitis. PMID- 15364011 TI - Milrinone efficiently potentiates insulin secretion induced by orally but not intravenously administered glucose in C57BL6J mice. AB - To study the effect of phosphodiesterase (PDE) 3 inhibition on plasma insulin and glucose levels, the selective PDE 3 inhibitor milrinone (0.25, 1.0, and 2.5 mg/kg) was given orally to anesthetized CL57Bl/6J mice 10 min before a gastric glucose gavage (150 mg/mouse). It was found that milrinone augmented the glucose mediated increase in plasma insulin at 1.0 and 2.5 mg/kg without, however, any improvement in glucose elimination. In contrast, when given 10 min before intravenous glucose (1 g/kg), milrinone (1 mg/kg) did not affect the insulin response to glucose. The increase in glucagon-like peptide-1 (GLP-1) levels after gastric glucose was not altered by milrinone. However, the PDE3 inhibitor augmented the insulin response to intravenous GLP-1 (2.8 nmol/kg). We therefore conclude that PDE3 inhibition by milrinone augments insulin secretion in vivo in mice after oral but not after intravenous glucose, which may be explained by enhanced response to the cAMP-dependent insulinotropic action of endogenously released GLP-1. PMID- 15364012 TI - Carbamazepine enhances brain production of kynurenic acid in vitro. AB - Disturbed formation of kynurenic acid, an endogenous antagonist of glutamate ionotropic receptors, might contribute to the pathogenesis of seizures. Here, the effect of anticonvulsant drug, carbamazepine on the production of kynurenic acid was studied. Carbamazepine (0.5-3 mM) enhanced kynurenic acid synthesis in rat cortical slices and also increased the activity of kynurenine aminotransferase (KAT) I at 0.1-3.0 mM concentration. Thus, anticonvulsant drugs, such as carbamazepine, might act partially via stimulation of kynurenic acid production. PMID- 15364013 TI - Influence of hippocampectomy on habituation, exploratory behavior, and spatial memory in rats. AB - Two frequently cited functions of the hippocampus are mediation of spatial memories and habituation. The present investigation employed head-shake response (HSR) as the habituated behavior in intact and bilaterally hippocampectomized rats. This HSR appears to be minimally influenced by spatial cues. These rats were further tested on two behavioral paradigms that make use of spatial cues, namely open field object exploration, and the Morris water maze. The results indicate that hippocampectomized rats revealed habituation of the HSR, but not to objects within the open field. In agreement with previous reports, hippocampectomized rats were severely impaired both in acquiring and recalling the location of the submerged platform in the Morris water maze task. In a separate experiment independent groups of rats were trained on one of these three paradigms, and tissues were collected from hippocampal, prefrontal, and piriform cortices for the measurement of matrix metalloproteinases (MMPs) as markers of neural plasticity. There were significant MMP-9 elevations in the prefrontal and piriform cortices of rats tested using the object exploration task, in the prefrontal and hippocampal cortices of rats that solved the Morris water maze task, but minimal MMP changes in any tissues taken from HSR habituated rats. These results question the hypothesis that habituation is solely mediated by the hippocampus in favor of a process that utilizes different brain structures and degrees of neural plasticity dependent upon task requirements. PMID- 15364014 TI - Microglial responses in the avascular quail retina following transection of the optic nerve. AB - This study was undertaken to investigate microglial responses in the avascular central nervous system using the quail retina that is known to be devoid of blood vessels. Following intraorbital optic nerve transection (ONT), the quail retina was examined immunohistochemically at various times up to 6 months. A few days after transection, microglia in the inner retinal layers revealed features of activation. Activated cells displayed an amoeboid shape and enhanced QH1 immunoreactivity. The numbers of these amoeboid cells were rapidly increased, first in the inner plexiform layer (IPL), and then in the ganglion cell/nerve fiber layer (GCL/NFL) of the retina where retrograde degenerating ganglion cell processes and perikarya were located. By 6 months after transection, microglia regained their resting morphology, and their cell counts returned to control levels. At early time points of microglial activation, numerous QH1+ amoeboid cells were observed along the vitreal surface of the pecten and retinal region adjacent to the insertion of the pecten, where some amoeboid cells were attached underneath the internal limiting membrane, and appeared to squeeze through the optic nerve fiber bundles. A considerable number of these amoeboid cells in the GCL/NFL and the IPL were labeled with PCNA, suggesting that active exogenous migration (from the pecten) and in situ proliferation of precursor cells contribute to the increase in microglial population of the degenerating retina. On the other hand, TUNEL-positive microglia appeared in the GCL/NFL at later time points indicate that the decrease of microglial numbers is in part due to apoptosis in these layers. Although some aspects of microglial activation in the avascular retina appear unique, their consequences were similar to those described in vascular retinae of mammals, a finding indicates that blood vessels are not a prerequisite for microglial activation, and microglial precursors could migrate long distance to reach the lesioned site, which is not accessible via blood vessels. Our data provide the first analysis of microglial activation in the avascular central nervous system (CNS), and suggest that the quail retina is a useful model for studies of microglial behavior in CNS. PMID- 15364015 TI - Aging alters norepinephrine release in the medial preoptic area in response to steroid priming in ovariectomized rats. AB - Changes in luteinizing hormone (LH) secretion that are observed in aging animals have been attributed to a reduction in hypothalamic norepinephrine (NE). The reason for the reduction in NE levels with aging is unclear. We hypothesized that the responsiveness of noradrenergic neurons to ovarian steroids is altered during aging. To test this, regularly cycling female Sprague-Dawley rats (young: 4-5 months old and middle age: 8-11 months old) were implanted with a push-pull cannula in the medial preoptic area (MPA) and ovariectomized bilaterally. On the 8th day after ovariectomy, they were injected with estrogen (30 microg/100 microl corn oil, s.c.) at 1000 h and on the 9th day they were implanted with a jugular catheter. On the 10th day they were injected with progesterone (2 mg/100 microl corn oil, s.c.) at 1000 h and subjected to push-pull perfusion. Perfusate samples from the MPA were collected at the rate of 10 microl/min every 30 min from 1300 to 1800 h and blood samples (0.3 ml) were collected hourly. The perfusate samples were analyzed for NE and dopamine (DA) concentrations using high performance liquid chromatography with electrochemical detection and serum LH levels were determined by RIA. In young animals, NE release (mean+/-S.E., pg/min) was 4.0+/ 1.1 pg/min at 1300 h and increased significantly (p<0.05) to 10.4+/-4.3 pg/min at 1500 h and remained elevated until 1600 h and then declined to 6.8+/-2.5 at 1730 h. In contrast, the increase in NE release occurred briefly in middle-aged animals and was delayed by an hour. LH patterns in both age groups followed the pattern in NE release. There was no change in the release of DA in both young and middle-aged animals. It is concluded that the altered responsiveness of noradrenergic neurons to steroid priming in middle-aged rats probably plays a critical role in the alterations seen in LH secretion in older animals. PMID- 15364016 TI - Upregulation of ceramide and its regulating mechanism in a rat model of chronic cerebral ischemia. AB - Ceramide is a key mediator of apoptosis, and is involved in the cellular stress response. We examined the alterations in the ceramide levels and their synthetic/degradative pathway in a rat model of chronic cerebral ischemia, in which ischemic white matter (WM) lesions occur in association with oligodendroglial cell apoptosis. Chronic cerebral ischemia was induced by clipping both common carotid arteries in male Wistar rats. After predetermined periods of 1, 3, 7 and 14 days, the animals were subjected to immunohistochemical and biochemical investigations for ceramide in the region containing the frontal cortex and corpus callosum (region 1), and the region containing the internal capsule and globus pallidus (region 2). After 14 days, the myelin was degraded in the corpus callosum, internal capsule and the optic tract in Kluver-Barrera staining. There was a significant increase in the ceramide level and the activity of its synthetic enzyme, acidic sphingomyelinase (SMase), whereas its degrading enzyme, glucosylceramide synthase (GCS), was downregulated in both regions 1 and 2 as compared to the sham-operated rats. Simultaneously, ceramide immunoreactive glia increased in number in the corpus callosum and the internal capsule after 3, 7 and 14 days. Double labeling for ceramide with glial fibrillary acidic protein but not with leukocyte common antigen indicated the astroglial nature of these glia. These findings indicate that chronic cerebral ischemia induces an increased ceramide level in astroglia as a result of downregulation of GCS and an upregulation of ASMase activity. PMID- 15364017 TI - A phenotype for the alpha 7 nicotinic acetylcholine receptor null mutant. AB - The alpha7 nicotinic acetylcholine receptor (nAChR) is heavily expressed in the mammalian brain. On a molecular level, the alpha7 nAChR may have a diversity of functions, but it is not known if these molecular events translate into phenotypes. The null mutant mouse is viable and generally normal. Here, we report a phenotype for the alpha7 nAChR null mutant mouse. The alpha7 nAChR is obligatory for the synchronization of an important biological rhythm, the female estrous cycle. The female null mutant mouse has asynchronous estrous cycles and a reduced number of surviving pups. Female null mutants also demonstrate a reliable diversity in phenotype, suggesting an interaction between environment and gene expression. Real-time RT-PCR measurements of the alpha7 mRNA expression in reproductive tissues of wild-type mice suggest that the ovulatory dysfunction in null mutants is probably central in origin. PMID- 15364018 TI - Bacterial endotoxin induces STAT3 activation in the mouse brain. AB - In the present study, we investigated regulatory mechanisms of bacterial endotoxin-induced STAT3 activation in the brain. Intraperitoneal injection of lipopolysaccharide (LPS) dose-dependently (0.5-5000 microg/kg) induced STAT3 phosphorylation in the hypothalamus. LPS-induced STAT3 phosphorylation was peaked at 2-4 h and declined there after. Moreover, intracerebroventricular injection of LPS induced STAT3 phosphorylation in the cortex and the hippocampus, indicating that central as well as peripheral LPS can act in the brain to induce STAT3 activation. Glucocorticoids are known to play a physiological role in the feedback inhibition of immune/inflammatory responses in the endocrine system. Interestingly, we observed no effect of dexamethasone on LPS-induced STAT3 phosphorylation in the hypothalamus. These findings point to the important role of STAT3 in the neuroimmune interaction of inflammation in the brain. PMID- 15364019 TI - Clozapine increases both acetylcholine and dopamine release in rat ventral hippocampus: role of 5-HT1A receptor agonism. AB - Atypical antipsychotic drugs (APDs) such as clozapine, but not the typical APD haloperidol, improve some aspects of cognition in schizophrenia. This advantage has been attributed, in part, to the ability of the atypical APDs to markedly increase acetylcholine (ACh) and dopamine (DA) release in rat medial prefrontal cortex (mPFC), while producing a minimal effect in the nucleus accumbens (NAC) or striatum. The atypical APD-induced preferential release of DA, but not ACh, in the mPFC is partially inhibited by the selective 5-HT(1A) antagonist WAY100635. However, little is known about these effects of atypical APDs in the ventral hippocampus (vHIP), another possible site of action of atypical APDs with regard to cognitive enhancement. The present study demonstrates that clozapine (10 mg/kg) comparably increases both ACh and DA release in the vHIP and mPFC. The increases in DA, but not ACh, release in both regions were partially attenuated by WAY100635 (0.2 mg/kg), which had no effect by itself on the release of either neurotransmitter in either region. Tetrodotoxin (TTX; 1 microM), a Na(+) channel blocker, in the perfusion medium, eliminated the clozapine (10 mg/kg)-induced ACh and DA release in the vHIP, indicating their neuronal origin. Haloperidol produced a slight increase in ACh release in the vHIP at 1 mg/kg, and DA release in the mPFC at 0.1 mg/kg. In conclusion, clozapine increases ACh and DA release in the vHIP and mPFC, whereas haloperidol has minimal effects on the release of these two neurotransmitters in either region. These differences may contribute, at least in part, to the superior ability of clozapine, compared to haloperidol, to improve cognition in schizophrenia. 5-HT(1A) agonism is important to the ability of clozapine and perhaps other atypical APDs to increase DA, but not ACh, release in the vHIP, as well as the mPFC. The role of hippocampus in the cognitive effects of atypical APDs warrants more intensive study. PMID- 15364020 TI - Multiple spatiotemporal patterns of dendritic Ca2+ signals in goldfish retinal amacrine cells. AB - Although it has been reported that dendritic neurotransmitter releases from amacrine cells are regulated by the intracellular Ca(2+) concentration ([Ca(2+)](i)), their spatiotemporal patterns are not well explained. Fast Ca(2+) imagings of amacrine cells in the horizontal slice preparation of goldfish retinas under whole-cell patch-clamp recordings were undertaken to better investigate the spatiotemporal patterns of dendritic [Ca(2+)](i). We found that amacrine cell dendrites showed inhomogeneous [Ca(2+)](i) increases in both Na(+) spiking cells and cells without Na(+) spikes. The spatiotemporal properties of inhomogeneous [Ca(2+)](i) increases were classified into three patterns: local, regional and global. Local [Ca(2+)](i) increases were observed in very discrete regions and appeared as discontinuous patches, presumably evoked by local excitatory postsynaptic potentials. Regional [Ca(2+)](i) increases were observed in either a single or a small number of dendrites, presumably reflecting the result of dendritic action potentials. Global [Ca(2+)](i) increases were observed in the entire dendrites of a cell and were mediated by Na(+) action potentials or multiple Na(+) action potentials riding on slow depolarization. Ca(2+)-mediated potentials also evoked global [Ca(2+)](i) increase in cells without Na(+) spikes. These spatiotemporal dynamics of dendritic Ca(2+) signals may reflect multiple modes of synaptic integration on the dendrites of amacrine cells. PMID- 15364021 TI - Noradrenaline-induced cation currents in isolated rat paratracheal ganglion neurons. AB - The actions of noradrenaline (NA) on the neurons acutely isolated from paratracheal ganglia of rats and the ionic mechanisms involved were studied with nystatin-perforated patch recording configuration. Under current-clamp conditions, application of 10 microM NA produced membrane depolarization followed by repetitive action potentials. NA evoked an inward cationic current under voltage-clamp conditions at a holding potential of -60 mV. Transient tail inward ('hump') current was also induced by washout of NA. The NA-induced current was reduced by extracellular Ca(2+) and Mg(2+), with half-maximal concentrations of 0.7 and 2.6 mM for Ca(2+) and Mg(2+), respectively. Phenylephrine, an alpha(1) adrenoceptor agonist, mimicked the NA-induced current, but the 'hump' current did not occur upon washout of phenylephrine. The NA-induced current was inhibited by prazosin and WB-4101, alpha(1)-adrenoceptor antagonists. In contrast, in the presence of yohimbine, an alpha(2)-adrenoceptor antagonist, the NA-induced current was potentiated and the washout of NA failed to evoke the 'hump' current. The pretreatment of paratracheal neurons with pertussis toxin also potentiated the NA-induced current. The NA-induced inward current was inhibited by pretreatment with U73122, a phospholipase C inhibitor, and xestospongin-C, a membrane-permeable IP(3) receptor antagonist. On the other hand, thapsigargin, BAPTA-AM and calmidazolium had no effect on the NA-induced current, suggesting that release of Ca(2+) from intracellular Ca(2+) stores via IP(3) receptors is not involved in the NA action. The cationic channels activated by NA play an important physiological role in neuronal membrane depolarization in rat paratracheal ganglia. PMID- 15364022 TI - Glial responses associated with dopaminergic striatal reinnervation following lesions of the rat substantia nigra. AB - Lesioning of dopaminergic substantia nigra pars compacta (SNpc) neurons leads to depletion of dopamine (DA) and dopaminergic axons in the dorsal striatum, followed by subsequent compensatory sprouting of dopaminergic fibers and striatal reinnervation. In this study, the response of striatal glia (microglia and astroglia) was compared with the degeneration and regeneration of dopaminergic axons following SNpc lesions. Following partial SNpc lesions, density of dopamine transporter (DAT) immunoreactive (-ir) terminals in the dorsal striatum returned to normal within 16 weeks of injury, suggesting that dopaminergic reinnervation of the striatum was complete. In conjunction, the glial responses in the dorsal striatum consisted of two peaks. The first peak in glial density occurred immediately after lesioning, peaking at 7 days, implying that it was likely to be associated with removal of debris from degenerating terminals. The second glial response commenced 8 weeks after lesioning and peaked some time after 16 weeks. The time of onset of the second peak suggests that it may be associated with the establishment of synapses rather than with axonal guidance. PMID- 15364023 TI - Delayed neuronal death and damage of GDNF family receptors in CA1 following focal cerebral ischemia. AB - Delayed neuronal death (DND) of pyramidal neurons in the CA1 and CA3 regions of the hippocampus has been extensively studied following global brain ischemia, whereas only little is known about DND in this highly vulnerable brain region after focal brain ischemia. In the present study, the distribution and time course of hippocampal neuronal apoptosis were studied following transient middle cerebral artery occlusion (MCAO) in rats 1, 3, 7, 14, and 30 days after the insult. In 60% of the animals, more than 90% of CA1 pyramidal neurons showed strong nick-end labeling (TUNEL) staining at day 3 with fragmentation and marginalization of the nuclei in approximately 40% of these cells. The number of TUNEL-positive cells decreased within the next days, but 30 days after MCAO, some apoptotic neurons were still present. Analysis of the expression of the glial cell line-derived neurotrophic factor (GDNF) and its receptors GFRalpha1, GFRalpha2, and GFRalpha3 using triple immunofluorescence and confocal laser scanning microscopy revealed that in all animals showing marked hippocampal DND, the neuronal staining for GFRalpha1, GFRalpha3, and GDNF decreased prior to the onset of TUNEL staining in CA1. After 7 days, some apoptotic neurons still expressed GFRalpha3, whereas only few showed GFRalpha1 immunoreactivity, indicating that GFRalpha1 may be beneficial for the survival of hippocampal neurons. The data suggest that reduced expression of GDNF and impairment of GFRalpha1/3 may contribute to hippocampal DND after focal brain ischemia. PMID- 15364024 TI - Essentiality of central GABAergic neuroactive steroid allopregnanolone for anticonvulsant action of fluoxetine against pentylenetetrazole-induced seizures in mice. AB - Fluoxetine, a selective serotonin reuptake inhibitor, is known to increase the cortical content of allopregnanolone (ALLO) without altering the level of other neurosteroids. In contrast to the proconvulsant effect of many antidepressants, fluoxetine exhibits anticonvulsant effects. The present study was undertaken to examine the role of ALLO in the anticonvulsant action of fluoxetine against pentylenetetrazole (PTZ)-induced seizures in mice. Prior administration of GABA(A) receptor agonist muscimol or neurosteroid ALLO or progesterone, a precursor of ALLO or neurosteroidogenic drugs like FGIN 1-27, an agonist at the mitochondrial diazepam binding inhibitor receptor (MDR) or metyrapone, an 11beta hydroxylase inhibitor, significantly potentiated the anticonvulsant effect of fluoxetine. In contrast, the effect of fluoxetine was counteracted by inhibition of the neurosteroid biosynthesis using drugs like 5alpha-reductase inhibitor, finasteride; 3beta-hydroxysteroid dehydrogenase inhibitor, trilostane; 3alpha hydroxysteroid dehydrogenase inhibitor, indomethacin; MDR antagonist, PK 11195; or the GABA(A) receptor antagonist, bicuculline. Further, bilateral adrenalectomy had no significant effect on the anticonvulsant action of fluoxetine, suggesting negligible contribution from peripheral steroidogenesis. The anticonvulsant effect of fluoxetine was partially abolished in 5,7-DHT treated mice, indicating that the effect may also, in part, be dependent on serotonergic transmission. Thus, our data indicate that increased synthesis of ALLO in CNS is a major factor that ultimately leads to anticonvulsant effects of fluoxetine against PTZ-induced seizures. PMID- 15364025 TI - Amitriptyline administration transforms tumor necrosis factor-alpha regulation of norepinephrine release in the brain. AB - The present study demonstrates that the mixed action antidepressant drug amitriptyline enhances norepinephrine (NE) release by transforming the nature of the response of neurons to both tumor necrosis factor-alpha (TNF) as well as to an alpha(2)-adrenergic agonist in an area of the central nervous system (CNS) rich in adrenergic neurons. Administration of the antidepressant drug amitriptyline for 1 day or 14 days to rats significantly increases TNF bioactivity in total homogenates of the locus coeruleus (LC) and the hippocampus as assessed by the WEHI-13VAR bioassay. Superfusion and electrical field stimulation of rat hippocampal brain slices were used to study the regulation of NE release. Exposure to TNF, as well as activation of the alpha(2)-adrenergic autoreceptor inhibits stimulation-evoked norepinephrine (NE) release from adrenergic neurons of the CNS from naive rats. Superfusion of hippocampal slices isolated from rats chronically (14 days) administered amitriptyline demonstrates that TNF inhibition of NE release is transformed, such that TNF facilitates NE release, dependent upon alpha(2)-adrenergic activation. Furthermore, chronic administration of amitriptyline increases stimulation-evoked NE release and decreases alpha(2)-adrenergic autoreceptor inhibition of NE release, an effect not observed with acute drug administration. These data support the hypothesis that chronic antidepressant drug administration, through regulation of TNF expression, transforms alpha(2)-adrenergic receptors such that they function to facilitate NE release, suggesting a mechanism of action of antidepressant drugs. PMID- 15364026 TI - Participation of histaminergic H1 and noradrenergic alpha 1 receptors in orexin A induced wakefulness in rats. AB - The participation of histaminergic H(1) and noradrenergic alpha(1) receptors in orexin A-induced wakefulness was studied by examining the sleep-wakefulness cycle in rats. Intracerebroventricular infusion of orexin A (1 nmol) caused an increase in the wakefulness state, while non-rapid eye movement sleep (NREM sleep) and rapid eye movement sleep (REM sleep) states were decreased. Prazosin (150 nmol) showed no significant antagonistic effect on the orexin A-induced increase in the wakefulness state and decrease in NREM and REM sleep. On the contrary, pyrilamine (150 nmol) was effective in antagonizing orexin A-induced increase in wakefulness and decrease in NREM sleep. When prazosin (150 nmol) and pyrilamine (150 nmol) were simultaneously perfused into the lateral ventricle, an almost complete antagonistic effect was observed with the increase in the wakefulness state and decrease in NREM sleep. Orexin A (1 nmol) caused a significant decrease in the histamine contents of the cortex, hippocampus and hypothalamus, whereas noradrenaline contents were decreased only in the hypothalamus. From these results, we concluded that the arousal effect induced by orexin A occurs through histaminergic H(1) and noradrenergic alpha(1) receptors, although participation of the H(1) receptor was more important than the alpha(1) receptor. PMID- 15364027 TI - Intranigral antagonism of neurokinin 1 and 3 receptors reduces intrastriatal dopamine D1 receptor-stimulated locomotion in the rat. AB - Stimulation of striatal dopamine (DA) D1 receptors increases the activity of the direct striatonigral pathway resulting in movement. While GABA has long been considered the primary effector of this pathway, co-released tachykinin peptides and their respective nigral tachykinin receptors are also in position to influence movement. Therefore, the present studies determined to what extent nigral tachykinin receptor subtypes contribute to striatal D1-mediated locomotion. Adult male Sprague-Dawley rats bearing chronic cannulae in the dorsal striatum and/or substantia nigra (SN) were tested for locomotor responses to various drug infusions. Unilateral intranigral infusions of the neurokinin-1 (NK1) antagonist LY306740 (0 and 50 nmol) but not the neurokinin-3 (NK3) antagonist SR142801 (0 and 50 nmol) led to ipsilateral rotations. Bilateral intrastriatal infusions of the full D1 agonist SKF 82958 (0, 1.2 and 12.0 nmol) dose-dependently increased locomotion. Prior bilateral intranigral infusions of LY306740 or SR142801 (0, 5.0 and 50 nmol) dose-dependently attenuated locomotor activity induced by intrastriatal SKF 82958 (12.0 nmol). These findings indicate that NK1, but not NK3, receptors within the SN may be tonically stimulated. However, activation of both nigral NK1 and NK3 receptors appears to be required for increased locomotion in response to striatal D1 receptor stimulation. PMID- 15364028 TI - Lysergic acid diethylamide and [-]-2,5-dimethoxy-4-methylamphetamine increase extracellular glutamate in rat prefrontal cortex. AB - The ability of hallucinogens to increase extracellular glutamate in the prefrontal cortex (PFC) was assessed by in vivo microdialysis. The hallucinogen lysergic acid diethylamide (LSD; 0.1 mg/kg, i.p.) caused a time-dependent increase in PFC glutamate that was blocked by the 5-HT(2A) antagonist M100907 (0.05 mg/kg, i.p.). Similarly, the 5-HT(2A/C) agonist [-]-2,5-dimethoxy-4 methylamphetamine (DOM; 0.6 mg/kg, i.p.), which is a phenethylamine hallucinogen, increased glutamate to 206% above saline-treated controls. When LSD (10 microM) was directly applied to the PFC by reverse dialysis, a rapid increase in PFC glutamate levels was observed. Glutamate levels in the PFC remained elevated after the drug infusion was discontinued. These data provide direct evidence in vivo for the hypothesis that an enhanced release of glutamate is a common mechanism in the action of hallucinogens. PMID- 15364029 TI - The brain-to-blood efflux transport of taurine and changes in the blood-brain barrier transport system by tumor necrosis factor-alpha. AB - The purpose of this study was to examine whether the efflux transport system for taurine from brain to blood is present at the blood-brain barrier (BBB) by using the brain efflux index (BEI) method and to determine whether the taurine transport system is regulated after central nervous system cell damage by tumor necrosis factor-alpha (TNF-alpha) in vivo. [(3)H]Taurine was microinjected into the parietal cortex area 2 of the rat brain, and was eliminated from the brain with an efflux transport rate of 1.22 x 10(-2) min(-1), and the process is saturable with a K(m) of 39.1 microM. This process was significantly inhibited by taurine transporter (TAUT) inhibitors, such as unlabeled taurine, beta-alanine, betaine, nipecotic acid and gamma-aminobutyric acid (GABA). In addition, the effect of tumor necrosis factor-alpha on [(3)H]taurine transport was investigated. [(3)H]Taurine uptake was increased and its efflux was reduced by pretreatment with tumor necrosis factor-alpha. Also, [(3)H]taurine efflux was reduced by tumor necrosis factor-alpha in a time- and dose-dependent manner. In conclusion, there is the efflux pump for taurine at the blood-brain barrier to reduce taurine concentration in the brain interstitial fluid, and this process was carrier mediated. In addition, the transport system for taurine through the blood-brain barrier was found to be regulated by tumor necrosis factor-alpha in vivo. PMID- 15364030 TI - Neuropeptide Y attenuates NMDA-induced phase shifts in the SCN of NPY Y1 receptor knockout mice in vitro. AB - Neuropeptide Y (NPY) blocks the effect of light on the mammalian circadian clock during the subjective night. The present study explores the role of the NPY Y1 receptor in this interaction. The effect of NPY when co-applied with NMDA, a glutamate agonist that can mimic the effect of light, was examined in NPY Y1-/- mice (background strain 129SVXBalb/c) using electrophysiology. Cells in the suprachiasmatic nucleus (SCN), the master circadian pacemaker, show a circadian rhythm in spontaneous firing rate that can be recorded in vitro. The results demonstrated that NPY attenuated the phase shifts to NMDA in both the Y1-/- mice and control mice, indicating that the Y1 receptor does not mediate the NPY blockade of photic-like phase shifts. The peak in frequency in the untreated control brain slices from Y1-/- mice was advanced by approximately 1 h as compared to the Y1+/+ mice. The Y1 receptor may contribute to a functional model of circadian rhythms, but apparently is not essential for the effects of NPY on photic phase shifts. PMID- 15364031 TI - The calcium influx pathway in rat olfactory ensheathing cells shows TRPC channel pharmacology. AB - Pharmacological characterisation of the calcium influx pathway in olfactory ensheathing cells (OECs) was performed using Indo-1 calcium microfluorometry. Our previous work has implicated this pathway in olfactory ensheathing cell support for regeneration of axons from adult CNS neurons. In high extracellular calcium (20 mM), cumulative concentration inhibition curves were generated for Lu(3+), Gd(3+) and econazole, giving IC(50)s of 0.09, 1.51 and 1.13 microM, respectively, and slope values that were not significantly different from unity. Combining these results with those obtained previously, an order of inhibitor potency was found to be Lu(3+)>La(3+)=econazole=Gd(3+)>1-[2-(4-methoxyphenyl)-2-[3-(4 methoxyphenyl)propoxy]ethyl-1H-imidazole hydrochloride (SKF96365)>Cd(2+). This profile most closely fits some members of the TRPC family of non-voltage gated calcium influx channels and may indicate that a TRP-mediated calcium influx plays a role in glial-neuronal interaction and axonal regeneration. PMID- 15364032 TI - Overexpression of the human beta-amyloid precursor protein downregulates cortistatin mRNA in PDAPP mice. AB - We measured preprocortistatin mRNA expression in young and aged transgenic (Tg) mice overexpressing the human beta-amyloid precursor protein (hbetaAPP) under the platelet-derived growth factor-beta promoter. Our findings suggest that the significant increase in hippocampal cortistatin mRNA expression during normal aging is significantly attenuated in Tg mice at an age known to exhibit beta amyloid protein (Abeta) deposition. These deficits in cortistatin expression may play a role in the deficits in hippocampal-dependent spatial learning and sleep/wake states previously demonstrated in aged Tg mice. PMID- 15364033 TI - Electrophysiological evidences for the contribution of NMDA receptors to the inhibition of clonidine on the RVLM presympathetic neurons. AB - The main objective of this study is to test the hypothesis that N-methyl-D aspartate (NMDA) receptors within the rostral ventrolateral medulla (RVLM) are involved in the inhibition of clonidine on the RVLM presympathetic neurons. Totally, 22 presympathetic neurons were recorded in anesthetized and paralyzed rats. The majority of these neurons (n=16 of 22) were significantly inhibited by iontophoretic (30 nA) clonidine, the other 6 neurons were insensitive to clonidine. In seven clonidine-sensitive neurons, iontophoretic clonidine (30 nA) antagonized the neuronal excitation of iontophoretic NMDA receptor agonist NMDA (20 nA). In remaining nine clonidine-sensitive neurons, iontophoretic NMDA receptor antagonist MK801 (60 nA) significantly attenuated the neuronal inhibition of iontophoretic (30 nA) clonidine. In conclusion, these results suggest that NMDA receptors contribute to the inhibition of clonidine on the RVLM presympathetic neurons. PMID- 15364034 TI - Flanking gene and genetic background problems in genetically manipulated mice. AB - Mice carrying engineered genetic modifications have become an indispensable tool in the study of gene functioning. The interpretation of results obtained with targeted mutants is not completely straightforward, however, because of genetic complications due to linkage and epistasis. Effects of closely linked genes flanking the targeted locus might sometimes be responsible for phenotypic changes ascribed to the null mutation. The effects of the latter might also be modified by the general genetic background. This review presents some examples and discusses some simple strategies to deal with these complications. PMID- 15364035 TI - Changes in cerebrospinal fluid neurochemistry during pregnancy. AB - BACKGROUND: Little is known about changes in brain function that may occur during pregnancy. Studies in rodents and sheep suggest that several brain neurotransmitter and neurohormonal systems known to modulate anxiety may be altered during pregnancy. METHODS: Cerebrospinal fluid (CSF) and plasma samples were obtained from 21 women (during weeks 38-39 of pregnancy) who were undergoing elective cesarean section and from 22 healthy nonpregnant women. RESULTS: The CSF levels of g-aminobutyric acid (GABA) and 3-methoxy-4-hydroxyphenylglycolwere reduced in pregnant women. There were no changes in CSF glutamate, 5 hydroxyindoleactic acid, and homovanillic acid. There was a large increase in CSF prolactin in pregnant women and also a trend toward an elevation in CSF oxytocin. Levels of prolactin, but not oxytocin, in CSF and plasma were correlated in pregnant women. CONCLUSIONS: These results suggest that pregnancy alters regulation of brain GABA, norepinephrine, and prolactin, which may play a role in changes in vulnerability to anxiety and depression during pregnancy and postpartum. Prolactin circulating in the bloodstream seems to be the major source of CSF prolactin during pregnancy. PMID- 15364036 TI - Smoking and respiratory irregularity in panic disorder. AB - BACKGROUND: The biological mechanisms underlying the link between smoking and panic attacks are unknown. Smoking might increase the risk of panic by impairing respiratory system function. METHODS: We evaluated the effect of smoking on respiratory irregularity in patients with panic disorder (PD) and healthy comparison subjects and the role of the respiratory disorders in this effect. We applied the Approximate Entropy index (ApEn), a nonlinear measure of irregularity, to study breath-by-breath baseline respiratory patterns in our sample. RESULTS: Both smoker and nonsmoker patients had more irregular respiratory patterns than healthy subjects. Smoker patients showed higher ApEn indices of baseline respiratory rate and tidal volume than nonsmoker patients (R = 5.4, df = 2,55, p < .01), whereas smoking in healthy subjects did not influence the regularity of respiratory patterns. Respiratory disorders did not account for the influence of smoking on respiratory irregularity. Smokers had more severe panic attacks than nonsmokers. CONCLUSIONS: Smoking may impair vulnerable respiratory function and act as disruptive factor on intrinsic baseline respiratory instability in patients with PD, possibly influencing the onset or maintenance of the disorder. PMID- 15364037 TI - Cross-sectional study of abnormal amygdala development in adolescents and young adults with bipolar disorder. AB - BACKGROUND: In vivo imaging studies in adult bipolar patients have suggested enlargement of the amygdala. It is not known whether this abnormality is already present early in the illness course or whether it develops later in life. We conducted a morphometric MRI study to examine the size of specific temporal lobe structures in adolescents and young adults with bipolar disorder and healthy control subjects, as well as their relationship with age, to examine possible neurodevelopmental abnormalities. METHODS: Subjects included 16 DSM-IV bipolar patients (16 +/- 3 years) and 21 healthy controls (mean age +/- SD = 17 +/- 4 years). Measures of amygdala, hippocampus, temporal gray matter, temporal lobe, and intracranial volumes (ICV) were obtained. RESULTS: There was a trend to smaller left amygdala volumes in patients (mean volumes +/- SD = 1.58 +/- .42 mL) versus control subjects (1.83 +/- .4 mL; F = 3.87, df = 1,32, p = .06). Bipolar patients did not show significant differences in right or left hippocampus, temporal lobe gray matter, temporal lobe, or right amygdala volumes (analysis of covariance, age, gender, and ICV as covariates, p > .05) compared with healthy control subjects. Furthermore, there was a direct correlation between left amygdala volumes and age (r =. 50, p = .047) in patients, whereas in healthy controls there was an inverse correlation (r = -.48, p = .03). CONCLUSIONS: The direct correlation between left amygdala volumes and age in bipolar patients, not present in healthy control subjects, may reflect abnormal developmental mechanisms in bipolar disorder. PMID- 15364038 TI - The -1438A/G polymorphism in the 5-hydroxytryptamine type 2A receptor gene affects promoter activity. AB - BACKGROUND: The -1438A/G single nucleotide polymorphism (SNP) lies just upstream of two alternative promoters for the 5-hydroxytryptamine type 2A (5-HT2A) receptor gene (HTR2A) and is in strong linkage disequilibrium with the 102T/C SNP. Both SNPs are associated with numerous psychiatric disorders and related phenotypes. A possible functional affect of the -1438A/G SNP might underlie associations of both linked SNPs with these neuropsychiatric disorders. A prior investigation into affects of this SNP on promoter function, lacking the more downstream promoter, found no significant difference with a reporter gene assay. METHODS: To investigate possible functional effects of -1438A/G on either promoter, two different reporter gene assays were used in three cell lines. RESULTS: Promoter activity was consistently detected that, in the presence of the SV40 enhancer, was significantly greater in the presence of the A allele relative to the G allele but only in cell lines that express endogenous HTR2A, suggesting that transcriptional factor(s) and the presence of both promoters might be necessary to elicit this effect. CONCLUSIONS: These findings show that the 1438A/G SNP has the potential to modulate HTR2A promoter activity and might be the functional variant responsible for the associations of both SNPs with many neuropsychiatric phenotypes. PMID- 15364039 TI - Meta-analysis of magnetic resonance imaging brain morphometry studies in bipolar disorder. AB - BACKGROUND: Several studies assessing volumetric measurements of regional brain structure in bipolar disorder have been published in recent years, but their results have been inconsistent. Our aim was to complete a meta-analysis of regional morphometry in bipolar disorder as assessed using magnetic resonance imaging (MRI). METHODS: We conducted a systematic literature search of MRI studies of bipolar disorder and identified studies which reported volume measurements in a selected number of regions. Twenty-six studies comprising volumetric measurements on up to 404 independent patients with bipolar disorder were included. A meta-analysis was carried out comparing the volumes of regions in bipolar disorder to comparison subjects using a random effects model. RESULTS: Patients with bipolar disorder had enlargement of the right lateral ventricle, but no other regional volumetric deviations which reached significance. Strong heterogeneity existed for several regions, including the third ventricle, left subgenual prefrontal cortex, bilateral amygdala and thalamus. CONCLUSIONS: Regional volume of most structures we studied is preserved in bipolar disorder as a whole, which was significantly associated only with right-sided ventricular enlargement. However the extensive heterogeneity detected indicates the need for further studies to establish if consistent regional brain volume deviation exists in bipolar disorder or in specific clinical subsets of the illness. PMID- 15364040 TI - Effect of vagus nerve stimulation on cerebrospinal fluid monoamine metabolites, norepinephrine, and gamma-aminobutyric acid concentrations in depressed patients. AB - BACKGROUND: Vagus nerve stimulation (VNS) has shown promising antidepressant effects in treatment-resistant depression, but the mechanisms of action are not known. Cerebrospinal fluid (CSF) studies in epilepsy patients show that VNS alters concentrations of monamines and gamma-aminobutyric acid (GABA), neurotransmitter systems possibly involved in the pathogenesis of depression. METHODS: Twenty-one adults with treatment-resistant, recurrent, or chronic major depression underwent standardized lumbar puncture for collection of 12 mL CSF on three separate but identical procedure days during participation in the VNS D-02 clinical trial. All subjects remained on stable regimens of mood medications. Collections were made at baseline (2 weeks after surgical implantation but before device activation), week 12 (end of the acute-phase study), and week 24. Cerebrospinal fluid concentrations of norepinephrine (NE), 5-hydroxyindoleacetic acid (5-HIAA), homovanillic acid (HVA), and 3-methoxy-4-hydroxyphenylglycol (MHPG) were determined with high-performance liquid chromatography. Concentrations of GABA were assayed with mass spectrometry. RESULTS: Comparison of sham versus active VNS revealed a significant (mean 21%) VNS-associated increase in CSF HVA. Mean CSF concentrations of NE, 5-HIAA, MHPG, and GABA did not change significantly. Higher baseline HVA/5-HIAA ratio predicted worse clinical outcome. CONCLUSIONS: Although several of the CSF neurochemical effects we observed in this VNS study were similar to those described in the literature for antidepressants and electroconvulsive therapy, the results do not suggest a putative antidepressant mechanism of action for VNS. PMID- 15364041 TI - Abnormalities in the dopamine system in schizophrenia may lie in altered levels of dopamine receptor-interacting proteins. AB - BACKGROUND: Dopamine receptor-interacting proteins constitute a part of the dopamine system that is involved in regulation of dopamine receptor-associated intracellular signaling. Previously, we demonstrated that two such proteins, the D1 receptor-interacting protein calcyon and the D2 receptor-interacting protein neuronal calcium sensor-1 (NCS-1), were elevated in the prefrontal cortex of schizophrenia cases from the Stanley Foundation Neuropathology Consortium. METHODS: The aim of this study was to confirm and expand these findings. We employed Western blot and real-time reverse transcriptase polymerase chain reaction analyses to compare prefrontal (area 46) and occipital (area 17) cortical levels of calcyon and NCS-1 proteins and mRNAs between schizophrenia (n = 37) and control (n = 30) cohorts from the Brain Collection of the Mount Sinai Medical School/Bronx Veterans Administration Medical Center. RESULTS: The schizophrenia cohort showed significant up-regulation of calcyon protein and message levels in both prefrontal and occipital cortical regions, both of which also displayed schizophrenia-associated up-regulation of NCS-1 message. Protein levels of NCS-1 were elevated only in the prefrontal cortex. All increases in protein levels were correlated with those of corresponding messages. Furthermore, schizophrenia-associated alterations in the levels of calcyon and NCS-1 messages were correlated. CONCLUSIONS: Up-regulation of calcyon and NCS-1 in the second schizophrenia cohort strengthens the proposition that abnormalities of the dopamine system in this disease may lie in altered levels of dopamine receptor interacting proteins. Also, up-regulation of both calcyon and NCS-1 in the cortex of schizophrenia patients can be attributed largely to an enhanced transcription or reduced degradation of their messages. Finally, our findings suggest that elevations in the expressions of calcyon and NCS-1 in schizophrenia may have the same underlying cause. PMID- 15364042 TI - Placebo-controlled trial of lamotrigine added to conventional and atypical antipsychotics in schizophrenia. AB - BACKGROUND: Lamotrigine, a novel anticonvulsant drug having modulatory effects on glutamatergic neurotransmission, improves mood and cognition parameters in bipolar disorder. Recent studies suggest that when added to clozapine, lamotrigine treatment may result in significant positive symptoms reductions in schizophrenia. Similar effects were not observed in an open trial in which lamotrigine was used as adjuvant to nonclozapine antipsychotics. METHODS: Thirty eight treatment-resistant schizophrenia inpatients receiving conventional and atypical antipsychotics enrolled in a 10-week, double-blind, placebo-controlled study, in which they were randomized in a 2:1 ratio to receive adjuvant treatment with lamotrigine, gradually titrated to a 400 mg/day dose, or placebo. Of these, 31 completed the trial. Measures of clinical efficacy and side effects were determined every other week. Serum levels of amino acids were assessed at the beginning and end of the study. RESULTS: In primary last observation carried forward analysis, no statistically significant between-group differences were observed; however, the completers' analyses revealed that lamotrigine treatment resulted in significant (p < or = .05) reductions in positive and general psychopathology symptoms, as measured by the Positive and Negative Syndrome Scale. No significant differences in lamotrigine effects were noted between conventional versus atypical antipsychotics. Lamotrigine treatment was well tolerated, and glutamate serum levels remained stable throughout the study. CONCLUSIONS: These preliminary findings 1) support the hypothesis that lamotrigine adjuvant treatment may improve positive symptoms and general psychopathology in schizophrenia, 2) suggest that beneficial effects may be achieved when lamotrigine is added to both conventional and atypical antipsychotics, and 3) warrant additional, larger scale trials. PMID- 15364043 TI - The relationship between volumetric brain changes and cognitive function: a family study on schizophrenia. AB - BACKGROUND: We examined the cerebral correlates of intelligence, memory, and executive processing in 56 patients with schizophrenia or schizoaffective disorder and 90 of their nonpsychotic relatives to establish whether the pattern of structure--function relationships in these two groups was different from that in 55 control subjects. METHODS: Magnetic resonance imaging data were acquired, and volumetric measurements were made for whole brain, prefrontal region, lateral ventricles, third ventricle, temporal lobes, hippocampi, and cerebellum. RESULTS: In the total sample, full intelligence quotient (IQ) and verbal IQ correlated with the volume of the whole brain and right hippocampus; the latter was also associated with performance IQ. Left hippocampal size was associated with verbal IQ and, in control subjects and nonpsychotic relatives only, with estimated full IQ. Delayed verbal memory was linked to cerebellar and inversely to left hippocampal volume. Discrepancies in the relationship pattern emerged in patients with schizophrenia between left hippocampus and measures of IQ and verbal memory. CONCLUSIONS: The latter data indicate a loss of a normal structure--function relationship in schizophrenia and might reflect a functional compensation occurring secondary to early neurodevelopmental impairment. PMID- 15364044 TI - A controlled study of brain structure in monozygotic twins concordant and discordant for schizophrenia. AB - BACKGROUND: We examined monozygotic twins concordant and discordant for schizophrenia to clarify the role of genetic and environmental factors in determining brain abnormalities. METHODS: Magnetic resonance imaging brain scans were obtained from 14 monozygotic twin pairs concordant and 10 monozygotic pairs discordant for schizophrenia, as well as 17 pairs of monozygotic control twins. Twenty-two discordant sibling-pairs and 56 pairs of unrelated control subjects were included to assess the extent of genetic control over these structures. RESULTS: Within-pair similarities for whole brain volume increased as pair members were more closely related genetically (monozygotic twins > siblings > unrelated control subjects). Schizophrenic twins, whether from concordant or discordant pairs, had smaller whole brain volumes than control twins. The probands of discordant pairs showed more abnormalities in hippocampal, third and lateral ventricular volumes than concordant twins. CONCLUSIONS: Whole brain volume is under high genetic control and smaller whole brain volume is a reflection of the genetic liability to develop schizophrenia. The variation in hippocampal and ventricular volumes within discordant monozygotic pairs indicates a role for environmental factors in determining these volume abnormalities in schizophrenia. Such factors may also underlie the more extensive morphometric deviations in patients from monozygotic discordant twins than in their counterparts from concordant twins. PMID- 15364046 TI - Ocular hypertension: defining risks and clinical options. PMID- 15364045 TI - Genetic and expression analyses of FZD3 in schizophrenia. AB - BACKGROUND: Wnt signaling plays important roles in neurodevelopmental processes. Frizzled is a receptor of Wnt protein, and the Frizzled 3 (FZD3) gene was recently reported to be associated with schizophrenia. Our study attempted to confirm associations between FZD3 and schizophrenia in Japanese family and case control samples. METHODS: Genetic associations were evaluated using family-based transmission tests (212 families, 643 subjects) and case--control analysis (540 schizophrenia patients, 540 control sample). Six single nucleotide polymorphisms (SNPs) on the FZD3 locus were genotyped, and levels of FZD3 mRNA expression in postmortem brains were examined. RESULTS: Neither family- nor population-based studies supported associations between FZD3 and schizophrenia. FZD3 expression was unaltered in schizophrenic brains. CONCLUSIONS: Although two prior studies have reported associations using limited numbers of SNPs on FZD3, our intensive study failed to support any major contribution of FZD3 to schizophrenia susceptibility. PMID- 15364047 TI - Applying an evidence-based approach to the management of patients with ocular hypertension: evaluating and synthesizing published evidence. AB - PURPOSE: The use of evidence-based medicine in managing patients with ocular hypertension has been constrained by the limited availability of high-quality data from controlled clinical trials and by limited formal training of clinicians in evaluating published evidence. This article will provide strategies and techniques to evaluate critically the quality of published research, synthesize the findings, and integrate published evidence in the care of patients with ocular hypertension. DESIGN: Review of the published literature and consensus of a panel of experts. METHODS: The published literature on evidence-based medicine was reviewed. Strategies and techniques in evaluating the quality of evidence and how to integrate evidence into the practice setting using the following three major questions were discussed by a panel of experts: Are the results of the study valid? What are the results? Do the study outcomes apply to individual patient care? RESULTS: Approaches to assessing study validity and the importance of considering the size of the treatment effect and precision of the effect size were detailed. The importance of using the "number-needed-to-treat" as a tool for translating results of clinical trials to individual patient care and the "number needed-to-harm" as a method of comparing the potential benefit and harm of a given therapy was acknowledged. CONCLUSION: As the volume of high-quality data increases, the information provided herein may help ophthalmologists apply evidence-based medicine techniques as they seek to optimize the care of individual patients with ocular hypertension. PMID- 15364048 TI - Glaucoma risk factor assessment and prevention: lessons from coronary heart disease. AB - PURPOSE: To determine whether multivariable risk factor assessment can be as successful in developing disease prevention strategies in patients with ocular hypertension as it has been in patients at risk for coronary heart disease (CHD). DESIGN: From a literature review of the evolution of the global risk assessment model for CHD, parallels are drawn to the evolution of a global risk assessment methodology for glaucoma. METHODS: This article summarizes the literature on global risk assessment and prevention of CHD in clinical practice, discusses the development of glaucoma risk assessment based on available trial and population data, and reviews the potential for prevention strategies founded on the cardiovascular disease model. RESULTS: To improve risk assessment in glaucoma and develop disease management strategies for patients with ocular hypertension, it is first necessary to identify and quantify levels of risk associated with factors that predict disease progression. In addition, the incidence of glaucoma and the average person's life expectancy based on his or her age are needed. Finally, it is necessary to quantify how long it takes to develop a visual defect that affects quality of life once a person develops glaucoma. CONCLUSION: The systematic application of epidemiologic data to CHD risk factor models provides insights into how global risk assessment can be incorporated into treatment recommendations for managing individuals with glaucoma. It is hoped that ophthalmologists can use the results of future clinical trials and long-term studies to develop disease prevention strategies in patients with ocular hypertension based on multivariable risk assessment. PMID- 15364049 TI - An evidence-based assessment of risk factors for the progression of ocular hypertension and glaucoma. AB - PURPOSE: To critically review the existing literature concerning risk factors for progression of ocular hypertension and glaucoma to assist in assigning levels of risk for individual patients. DESIGN: Evidence-based review. METHODS: A panel of physicians specializing in treatment of glaucoma patients was convened to critically analyze published population-based studies of ocular hypertension and glaucoma progression. The strength of evidence in support of reported risk factors was weighed. RESULTS: Many putative risk factors for progression of ocular hypertension or glaucoma have been reported in the literature. The risk factors most strongly supported by evidence are higher intraocular pressure (IOP), greater cup-to-disk ratio, thinner central corneal measurement, and older age. Black race does not appear to be an independent risk factor, although black individuals tend to have thinner corneas, greater cup-to-disk ratios, and higher IOP, which increase their risk. The limited number of studies in which other suspected risk factors are reported prevents drawing firm conclusions about their importance at this time. CONCLUSIONS: Only a subset of patients with ocular hypertension will eventually develop glaucoma. Decisions regarding the implementation and extent of therapy for ocular hypertension can be difficult and require an understanding of the relative importance of risk factors for progression. This review discusses the strength of evidence supporting reported risk factors and may be useful in assessing the risk for progression of individual patients. PMID- 15364050 TI - Striking a balance: inter-kingdom cell-to-cell signaling, friendship or war? AB - There is a growing body of evidence suggesting that bacterial signaling molecules are involved in cross-kingdom communication with eukaryotic cells. Bacterial signaling molecules can enter eukaryotic cells and appear to modulate immune responses to bacterial pathogens. Recently, an enticing study demonstrated that mammalian airway epithelial cells have the ability to inactivate these bacterial signaling molecules, suggesting that this activity might have a role in the host innate response against bacterial infections. PMID- 15364051 TI - More sympathy for autoimmunity with neuropeptide Y? AB - Substantial evidence indicates a dysfunctional communication between the sympathetic nervous system and the immune system in Th1-mediated autoimmune diseases, such as rheumatoid arthritis and multiple sclerosis. In this Opinion, we propose that the sympathetic regulation of immunity is not only mediated by catecholamines but also involves neuropeptide Y (NPY), an additional postganglionic SNS transmitter that is shown to modulate various immunological functions in vitro and in vivo. Based on recent experimental findings, we believe that a more precise understanding of the role of NPY in the regulation of autoimmune Th1 cells will provide novel insights into the neuroimmunological basis of autoimmunity. PMID- 15364052 TI - Smad7 in TGF-beta-mediated negative regulation of gut inflammation. AB - Mice with targeted disruptions of the transforming growth factor-beta1 (TGF beta1) gene or TGF-beta1 intracellular signalling pathways develop intestinal inflammation. Conversely, TGF-beta1-producing regulatory T cells protect against experimental colitis. Paradoxically, however, TGF-beta1 production is high in the gut of patients with chronic inflammatory intestinal disease, and yet inflammation proceeds unchecked. Here we discuss the functional role of Smad7, an intracellular inhibitor of TGF-beta1 signalling, in the control of gut inflammation by TGF-beta1. In particular, we delineate a scenario in which the high expression of Smad7 in inflammatory cells renders them unresponsive to TGF beta1 and propose that control of Smad7, not TGF-beta1 production, is a key determinant in understanding how TGF-beta1 negatively regulates gut inflammation. PMID- 15364053 TI - Tolerance in mixed chimerism - a role for regulatory cells? AB - The establishment of mixed hematopoietic chimerism induces life-long donor specific organ graft tolerance while obviating the need for chronic immunosuppression. Recent advances have dramatically reduced the conditioning toxicity required to achieve mixed chimerism. We argue that the achievement of high levels of donor chimerism ensures life-long deletion of donor-reactive T cells, precluding and obviating the need for regulatory mechanisms in the maintenance of tolerance. However, in situations where high levels of donor chimerism cannot be established or sustained, control of immune responsiveness can be achieved through additional mechanisms, including regulatory T cells. PMID- 15364054 TI - Can we really learn from model pathogens? AB - Cytolytic T (Tc) lymphocytes are the first order response of the adaptive immune system in the recovery from primary viral infections. These effector cells execute their function either by direct cytotoxicity through the Fas or perforin pathway and/or by the release of cytokines that either directly or indirectly exert antiviral activity. Mice respond to infection by closely related viruses with a vigorous Tc response, which is characterized by extensive crossreactivity on target cells infected with these viruses. However, the action of these cells can be beneficial, detrimental or neutral. From our current state of knowledge, no generalizations as to protective or detrimental effects of cytolytic effector functions in recovery from virus infections can be made. Thus, virus-host immune interactions have to be assessed individually and cannot be generalized. PMID- 15364055 TI - Signal regulators in FcR-mediated activation of leukocytes? AB - Leukocyte membrane receptors for the Fc portion of Igs (FcRs) link antigen recognition by antibodies to effector functions involved in immune phenomena, from pathogen elimination to autoimmunity. Moreover, they also signal for the synthesis and secretion of cytokines and chemokines, thus having a role in immune homeostasis. Even though the structural and functional similarities between FcRs and the clonotypic antigen receptors of lymphocytes (the T-cell receptor and B cell receptor) are well established, participation of regulatory membrane molecules in leukocyte activation by FcRs has rarely been considered. Here, we summarize evidence demonstrating that FcR-mediated signaling could be modulated by other membrane molecules (signal regulators), and propose that comprehension of this phenomenon is essential for understanding the functions of FcRs, knowledge of which could then be used for therapeutic interventions. PMID- 15364056 TI - Interactions between the innate immune and blood coagulation systems. AB - Blood coagulation and inflammation are universal responses to infection and there is crosstalk between inflammation and coagulation that can either amplify or dampen the responses. Loss of appropriate interactions between these systems probably contributes to morbidity and mortality in infectious diseases. For instance, inflammatory cytokines and leukocyte elastase can downregulate natural anticoagulant proteins that help to maintain endothelial-cell integrity, control clotting, inhibit vasoactive peptides and dampen leukocyte infiltration into the vessel wall. This Review will summarize our current understanding of the mechanisms involved in the crosstalk between these two important systems. PMID- 15364057 TI - Molecular interactions regulate BCR signal inhibition by CD22 and CD72. AB - The inhibitory coreceptors CD22 and CD72 downmodulate B-cell receptor (BCR) signaling and function as a molecular switch, determining whether antigen stimulated B cells undergo apoptosis or proliferation. These coreceptors carry an intrinsic property for associating with the BCR, and this association is crucial for the initiation of signal inhibition through phosphorylation of these coreceptors by BCR-associated kinases. Recent findings have demonstrated that signal inhibition by these coreceptors is regulated by ligands for the coreceptors and by molecules binding to the coreceptors or the BCR. Moreover, signal inhibition by CD22 depends on the BCR isotype. These findings suggest a dynamic regulation of these coreceptors through molecular interactions on the B cell surface. PMID- 15364058 TI - C1q and tumor necrosis factor superfamily: modularity and versatility. AB - C1q is the target recognition protein of the classical complement pathway and a major connecting link between innate and acquired immunity. As a charge pattern recognition molecule of innate immunity, C1q can engage a broad range of ligands via its globular (gC1q) domain and modulate immune cells, probably via its collagen region. The gC1q signature domain, also found in many non-complement proteins, has a compact jelly-roll beta-sandwich fold similar to that of the multifunctional tumor necrosis factor (TNF) ligand family. The members of this newly designated 'C1q and TNF superfamily' are involved in processes as diverse as host defense, inflammation, apoptosis, autoimmunity, cell differentiation, organogenesis, hibernation and insulin-resistant obesity. This review is an attempt to draw structural and functional parallels between the members of the C1q and TNF superfamily. PMID- 15364059 TI - Breakthrough antibody therapy for lethal cancer. PMID- 15364060 TI - Creatine as a neuroprotector. PMID- 15364061 TI - Gene therapy for muscular disease. PMID- 15364062 TI - Stephen Neidle on cancer therapy and G-quadruplex inhibitors. Interview by Joanna De Souza. AB - Stephen Neidle was educated at Imperial College, London, where he graduated in chemistry and then proceeded to do a PhD in crystallography. After a period as an ICI Fellow, he joined the Biophysics Department at King's College, which ignited his interest in nucleic acid structural studies. He was appointed as one of the first Cancer Research Campaign Career Development Awardees, becoming a Life Fellow on moving to the Institute of Cancer Research. He was appointed to the Chair of Biophysics at the Institute of Cancer Research in 1990, and moved to the new Chair of Chemical Biology at the School of Pharmacy in the University of London in 2002, where he also directs the Cancer Research UK Biomolecular Structure Group. He is currently Chairman of the Chemical Biology Forum of the Royal Society of Chemistry, which is involved in developing the interface between chemistry and the life sciences. He will shortly assume the Directorship of the newly-established Centre for Cancer Medicines at the School. Stephen Neidle has received several awards for his work on drug-nucleic acid recognition and drug design, including the 2000 prize of the Biological and Medicinal Chemistry Sector of the Royal Society of Chemistry, and its 2002 Interdisciplinary Award. He was the 2004 Paul Ehrlich Lecturer of the French Societe de Chimie Therapeutique, and was recently awarded the 2004 Aventis Prize in Medicinal Chemistry. PMID- 15364063 TI - Therapeutic control of free radicals. PMID- 15364064 TI - Drug discovery in the HIF pathway: where do we go now? PMID- 15364065 TI - Opening the bottlenecks. PMID- 15364066 TI - Target discovery in metabolic disease. AB - The prevalence of metabolic diseases is taking on epidemic proportions and poses a serious threat to human health. Current treatment options have proven insufficient to cope with obesity and diabetes because they rarely restore normal metabolism and thus leave patients exposed to life-threatening complications. Successful management of these diseases depends on novel, improved therapeutic strategies targeting early intervention in disease progression. Discovery of novel metabolic disease targets has been hampered by the complexity of contributing environmental and genetic factors, as well as the need for potent but safe treatments suitable for chronic diseases. Genomic approaches are excellent tools to manage genetic complexity and have been applied successfully to identify candidate target genes that will lead to the development of novel therapies for metabolic diseases. PMID- 15364067 TI - Bioinformatics and cancer target discovery. AB - The convergence of genomic technologies and the development of drugs designed against specific molecular targets provides many opportunities for using bioinformatics to bridge the gap between biological knowledge and clinical therapy. Identifying genes that have properties similar to known targets is conceptually straightforward. Additionally, genes can be linked to cancer via recurrent genomic or genetic abnormalities. Finally, by integrating large and disparate datasets, gene-level distinctions can be made between the different biological states that the data represents. These bioinformatics approaches and their associated methodologies, which can be applied across a range of technologies, facilitate the rapid identification of new target leads for further experimental validation. PMID- 15364068 TI - Biomaterial challenges and approaches to stem cell use in bone reconstructive surgery. AB - As life expectancy increases, so does the need to treat large bone defects. New biomaterials combined with osteogenic cells are now being developed as an alternative to autogenous bone grafts. The goal is to make the stem cells adhere to the scaffold, and then grow to differentiate into functional osteogenic cells and organize into healthy bone as the scaffold degrades. Decisive improvements have been made in the fields of stem cell biology, 3-D scaffold fabrication and tissue engineering, but the ideal bone substitute that fulfils all functional and safety requirements has yet to be developed. PMID- 15364073 TI - Optimum doses of Mg and P salts for precipitating ammonia into struvite crystals in aerobic composting. AB - It was previously reported that struvite crystals could be formed in the aerobic composting reaction provided that Mg and P salts are added [Bioresource Technology 79 (2001) 129]. The formation of struvite crystals significantly reduced gaseous loss of ammonia and resulted in substantial increase in the ammonia content in the compost, attaining 1.5%. In this context, the present study was conducted to determine the optimal doses of Mg and P salts for struvite crystallization. It was found that cumulative ammonia production was about 33-36% of the initial total nitrogen in the aerobic composting reaction, irrespective of the amounts of Mg and P salts added. The theoretical doses for complete conversion of ammonia into struvite crystals seemed to be about 33-36% of the initial nitrogen. The addition of Mg and P salts at this level, however, caused adverse effects on the degradation of organic materials. Therefore, it was concluded that the optimal doses of Mg and P salts should be about 20% of the initial nitrogen in the compost mixture not to cause any harmful effects on the composting reaction. PMID- 15364074 TI - Biological nutrient removal by a sequencing batch reactor (SBR) using an internal organic carbon source in digested piggery wastewater. AB - Experiments in a lab-scale SBR were conducted to demonstrate the feasibility of using an internal carbon source (non-digested pig manure) for biological nitrogen and phosphorus removal in digested piggery wastewater. The internal C-source used for denitrification had similar effects to acetate. 99.8% of nitrogen and 97.8% of phosphate were removed in the SBR, from an initial content in the feed of 900 mg/l ammonia and 90 mg/l phosphate. PMID- 15364075 TI - Environmental effects and desorption characteristics on heavy metal removal using carboxylated alginic acid. AB - Effects of ionic strength and organic materials on copper ion uptake capacity using carboxylated alginic acid which showed very high metal ion uptake capacity were investigated. The ionic strength only had a slight effect on the decrease of copper ion uptake capacity regardless of NaCl concentration. And, the effect of organic materials such as NTA (nitrilotriaceticacid) and sodium hypophosphite on the copper ion uptake capacity was negligible. When the lead ion adsorbed on carboxylated alginic acid was desorbed by NTA, which showed high desorption efficiency, the best optimum concentration of NTA was about 0.01 M. Also desorption efficiency decreased, however, concentration factor increased as S/L ratio which is defined as the ratio of adding amount of adsorbed and volume of desorbing agent increased. In sequential adsorption and desorption cycles, the lead uptake capacity on carboxylated alginic acid was relatively maintained through cycles 1-5. PMID- 15364076 TI - The effect of xylanase on lignocellulosic components during the bleaching of wood pulps. AB - HPLC, SEM and XRD techniques have been proposed as methods for ascertaining the changes occurring in polysaccharides (cellulose and xylans) and fibres during the xylanase bleaching processes. TCF and ECF bleached pulps with and without enzyme pretreatment were analysed. The ratio of carbohydrates present in the pulp, observation of changes occurring in the surface of the fibres and the crystallinity and accessibility of the bleached fibres were determinated. These characteristics have been related with pulp properties. Xylan content decreased when pulp was bleached. Xylanase treatment substantially reduced the xylose content present in pulp, measured by HPLC after the hydrolysis method of the sample. Morphological changes in the fibres occurred when the enzymatic treatment was applied. Bleaching increased the crystallinity of the pulp and enzyme pretreatment also affected the crystallinity of cellulose fibres PMID- 15364077 TI - Effects of synthetic polymer on the filamentous bacteria in activated sludge. AB - Filamentous bulking is one of the solid-liquid separation problems always seen in activated sludge process. The addition of synthetic polymer is always one of the popular ways for the treatment plant operator to immediately solve the poor sludge settling problem. Therefore, it may be interesting to understand the effects of synthetic polymer on the filamentous bacteria in activated sludge. In this study, synthetic polymer was applied to a lab-scale wastewater treatment system with the filamentous bulking problem. The population structure of filamentous bacteria and sludge characteristics were investigated under different conditions. When synthetic polymer was added into the system, it was found that poor sludge settleability caused by filamentous bulking was temporarily solved and filamentous branches growing outside the flocs were damaged or inhibited. However, filamentous growth was still observed inside the flocs. After the addition of polymer was halted, filamentous branches extended out of the flocs immediately. Very serious filamentous bulking occurred and sludge settleability became much worse than that occurring before the addition of polymer. And, it took several weeks for the system to return to normal operation. PMID- 15364078 TI - Removal of volatile fatty acids with immobilized Rhodococcus sp. B261. AB - The removal of aqueous volatile fatty acids (VFA) in wastewater and spoiled waste foods by immobilized Rhodococcus sp. B261 was investigated. The n-valeric acid (0.5%) was completely removed within 25 h under the following conditions; solution pH, 8.0; air flow rate, 0.2 l/min; superficial velocity, 0.96 h(-1); temperature, 37 degrees C. Under the optimized conditions, the acetic (8525 ppm), propionic (7310 ppm) and n-butyric (4360 ppm) except n-valeric (2572 ppm) acids from the wastewater were completely removed by immobilized Rhodococcus sp. B261 in 24 h. The acetic (7810 ppm), propionic (8942 ppm) and butyric (5730 ppm) acids from the solution of spoiled waste-foods were effectively removed by immobilized Rhodococcus sp. B261 from 48 h within 60 h but n-valeric acid (3625 ppm) took 72 h. PMID- 15364079 TI - Processing cotton gin trash to enhance in vitro dry matter digestibility in reduced time. AB - Cotton gin trash (CGT) in the raw form is poorly digested by ruminants due to lignocellulosic complexes. These structures must be broken down before adequate digestion can occur. This may be performed by physical and/or chemical means. Two methods for improving digestibility are particle size reduction and/or treatment with sodium hydroxide (NaOH). To evaluate the effectiveness of each method, three experiments were performed in which different CGT types were tested. Each type represented trash from a particular cleaning stage in the cotton ginning process. First, each type was ground with a knife-type grinding mill using screen sizes 0.5, 1.0, and 2.0 mm. For the second experiment, particle size was held constant at 2 mm, and all CGT types were treated with 4% and 6% NaOH (w/w) at room temperature. An agitation cycle of 5 min on and 10 min off was used, with the total mixing time being 4 h. Lastly, particle size and NaOH concentration were held constant, and treatments were performed at room temperature, 40 degrees C, and 50 degrees C. The total mixing times were 2 and 3 h for 50 and 40 degrees C, respectively. For all experiments two subsamples of each treatment were tested for in vitro dry matter digestibility (IVDMD). From grinding alone, digestibility increased as particle size decreased. Grinding to 0.5 mm resulted in an average IVDMD of 47.8% while grinding to 2.0 mm resulted in an average IVDMD of only 33.8%. Digestibility also improved with a greater NaOH concentration. An average in vitro digestibility of 70.5% was achieved with 6% NaOH (w/w) treatment, essentially doubling that of the raw CGT. Increasing the reaction temperature did not result in increased digestibility because the mixture dried out, with a consequent reduction in chemical distribution and uniformity in heat transfer. There are still chemical residues in the CGT, and elimination/reduction of these is an issue that needs to be addressed in further research. PMID- 15364080 TI - Biotreatment of wastewater using aquatic invertebrates, Daphnia magna and Paramecium caudatum. AB - A number of major changes have occurred over the past few years, which give cause for a re-examination of conventional wastewater treatment methods. Among these are growing problems of worldwide energy and food shortages and nutrients not removed by conventional secondary processes causing algal blooms and other problems in the receiving waters. The global increase in wastewater calls for innovative low cost technology approaches to its recycling. Biotreatment systems, utilizing living organisms are receiving growing attention since they are ecologically sound, cheap and applicable in areas without land constraints. Filter feeders (both invertebrates and vertebrates) are promising in this area since they can remove suspended organic matter and bacteria, even in the size range of microns. In the present study biological treatment of municipal wastewater using two invertebrates--Paramecium caudatum, a protozoan and Daphnia magna, a cladoceran was investigated. Analysis at pre-experimental and post experimental stages revealed the potential of these species in abatement of water pollution. D. magna was more efficient than P. caudatum in laboratory-scale studies. PMID- 15364081 TI - Removal of chlorophenols from aquatic systems using the dried and dead fungus Pleurotus sajor caju. AB - In this study, the potential use of the fungus Pleurotus sajor caju to remove phenols (i.e., phenol, o-chlorophenol, p-chlorophenol and 2,4,6-trichlorophenol) from aqueous solutions was evaluated. Biosorption of phenol or chlorophenols reached equilibrium in 4 h. The maximum adsorptions of phenol and chlorophenols onto the Pleurotus sajor caju were 0.95 mmol/g for phenol, 1.24 mmol/g for o chlorophenol, 1.47 mmol/g for p-chlorophenol and 1.89 mmol/g for 2,4,6 trichlorophenol. The affinity order was as follows: 2,4,6-trichlorophenol> p chlorophenol> o-chlorophenol>phenol. Phenol and chlorophenols bindings onto Pleurotus sajor caju were clearly pH dependent. The adsorption of phenol and chlorophenols increased with increasing pH. Desorption was achieved using methanol solution (30%, v/v). Pleurotus sajor caju biomass is suitable for reuse for more than five cycles without noticeable loss of adsorption capacity. PMID- 15364082 TI - Joint effect of nitrogen sources and B vitamin supplementation of date juice on lactic acid production by Lactobacillus casei subsp. rhamnosus. AB - The use of date juice as a substrate for lactic acid production was investigated. Various nitrogen sources were compared with yeast extract for efficient lactic acid production by Lactobacillus casei subsp. rhamnosus. Among different nitrogen sources added to date juice (yeast extract, ammonium sulfate, tryptic soy, urea, peptone, and casein hydrolysate), yeast extract was the most efficient. The effect of yeast extract could have been due to its B vitamin content. The addition of five B vitamins at less than 25 mg/l to date juice with any nitrogen source enhanced lactic acid production to some extent, except for date juice with yeast extract or urea or peptone. The most significant increase was obtained with ammonium sulfate. Half of the yeast extract content (10 g/l) in a supplemented date juice could be replaced by a mixture of B vitamins at less than 25 mg/l, and ammonium sulfate at 2.6 g/l with no significant decrease in lactic acid production. PMID- 15364083 TI - Bio-degradation of olive oil husks in composting aerated piles. AB - In this study, the composting performance of two olive oil husk piles was compared using two different aeration processes: aeration by mechanical turning or by forced air-injection. The results showed that after the maturation phase both piles had a similar chemical composition and the same level of organic matter degradation (around 55%). However, the time necessary to reach the thermophilic phase was shorter for the turned pile (2 months in comparison to 3 months for the air-injected pile) and the humification degree achieved was slightly higher (94% versus 83%). Both piles showed the presence of an active microbial community, with an increase by 2-3 orders of magnitude in the number of cultured microbial colonies during the composting process. No significant difference in quantitative or qualitative terms was found in the microbial populations of both piles, nor was a clear succession between a mesophilic and a thermophilic population observed. In terms of industrial application, the mechanical turning process seems to be better since it is a lower energy and time consuming process. PMID- 15364084 TI - Alkaline pulping with additives of date palm rachis and leaves from Sudan. AB - Soda-anthraquinone (soda-AQ), alkaline sulphite-anthraquinone (AS-AQ) and alkaline sulphite-anthraquinone-methanol (ASAM) pulping of date palm rachis and leaves from Sudan was carried under different conditions, and pulps with variable yields and mechanical properties were obtained. The date palm rachis gave best yields and mechanical properties with the AS-AQ or the ASAM process, while the leaves were best pulped with the soda method with low yield, but very good strength properties. Blending with 10% and 30% kenaf bark pulp was beneficial, especially for the AS-AQ pulps. Totally chlorine free (TCF) bleached rachis pulps were obtained of high brightness and strength properties suitable for use in writing and printing papers. PMID- 15364085 TI - Anaerobic treatment of wastewater with high suspended solids from a bulk drug industry using fixed film reactor (AFFR). AB - Studies were carried out on the treatment of wastewater from a bulk drug industry using an anaerobic fixed film reactor (AFFR) designed and fabricated in the laboratory. The chemical oxygen demand (COD) and total dissolved solids (TDS) of the wastewater were found to be very high with low biochemical oxygen demand (BOD) to COD ratio and high total suspended solid (TSS) concentration. Acclimatization of seed consortia and startup of the reactor was carried out by directly using the wastewater, which resulted in reducing the period of startup to 30 days. The reactor was studied at different organic loading rates (OLR) and it was found that the optimum OLR was 10 kg COD/m(3)/day. The wastewater under investigation, which had a considerable quantity of SS, was treated anaerobically without any pretreatment. COD and BOD of the reactor outlet wastewater were monitored and at steady state and optimum OLR 60-70% of COD and 80-90% of BOD were removed. The reactor was subjected to organic shock loads at two different OLR and the reaction could withstand the shocks and performance could be restored to normalcy at that OLR. The results obtained indicated that AFFR could be used efficiently for the treatment of wastewater from a bulk drug industry having high COD, TDS and TSS. PMID- 15364086 TI - Evidence of chemical reactions between di- and poly-glycidyl ether resins and tannins isolated from Pinus radiata D. Don bark. AB - This work evaluates the feasibility of reacting tannins isolated from Pinus radiata D. Don bark with epoxide resins of the diglycidyl and polyglycidyl ether type. To this end, gel times of aqueous tannin dispersions (40% w/w) with every one of nine selected resins (5% w/w), at previously established pH values (initial equal to 3.3, 4, 7 and 10), have been determined. Products of these reactions were analyzed by FT-IR spectroscopy, and the results were compared with those obtained from tannin-p-formaldehyde and (+)-catechin-p-formaldehyde systems, at the same pH values. Their mechanical properties were evaluated, by dynamic mechanical analysis, at two pH values (3.3 and 10). In general, it was concluded that tannin-epoxide resin systems behave similarly to tannin paraformaldehyde systems, especially at basic pH values. PMID- 15364087 TI - Biosorption of cadmium and lead ions by ethanol treated waste baker's yeast biomass. AB - The biosorption of cadmium and lead ions from artificial aqueous solutions using waste baker's yeast biomass was investigated. The yeast cells were treated with caustic, ethanol and heat for increasing their biosorption capacity and the highest metal uptake values (15.63 and 17.49 mg g(-1) for Cd(2+) and Pb(2+), respectively) were obtained by ethanol treated yeast cells. The effect of initial metal concentration and pH on biosorption by ethanol treated yeast was studied. The Langmuir model and Freundlich equation were applied to the experimental data and the Langmuir model was found to be in better correlation with the experimental data. The maximum metal uptake values (qmax, mg g(-1)) were found as 31.75 and 60.24 for Cd(2+) and Pb(2+), respectively. Competitive biosorption experiments were performed with Cd(2+) and Pb(2+) together with Cu(2+) and the competitive biosorption capacities of the yeast biomass for all metal ions were found to be lower than in non-competitive conditions. PMID- 15364088 TI - Vermicomposting of cattle and goat manures by Eisenia foetida and their growth and reproduction performance. AB - Vermicomposting is commonly adopted for the treatment of livestock organic wastes. In the present study, two types of livestock manure were used for culturing of the earthworm, Eisenia foetida. Each treatment group consisted of six replicates and worm vermicasts were examined after 5 weeks. The concentrations of total C, P and K in goat manure vermicasts were higher than those in cattle manure vermicasts. Cattle vermicasts had a higher N content than goat vermicasts but the C:N ratio of fresh manure was higher than that of vermicasts for both materials. Earthworm biomass and reproductive performance, in terms of number of worms after 5 weeks of experiment, were higher in cattle manure than in goat manure. The cocoon production per worm in cattle manure was higher than in goat manure. However, the hatchability of cocoons was not affected by manure treatments. In conclusion, cattle manure provided a more nutritious and friendly environment to the earthworms than goat manure. PMID- 15364089 TI - Advanced start-up of anaerobic attached film expanded bed reactor by pre-aeration of biofilm carrier. AB - The start-up and performance of the anaerobic attached film expanded bed (AAFEB) reactor with pre-aeration of carrier were investigated. The carriers of the reactors had been aerated for 10 days before they were put into the AAFEB reactors. The results indicated that the reactors advance the start-up by 15 days, and maintain higher efficiency when they were subjected to organic and hydraulic loading shock, but during steady-state operation, the reactors did not show better performance than the control reactors without pre-aeration of carrier. The thicker biofilm and higher biomass concentration of the reactors with pre-aeration were observed during the start-up period, but the difference between two types of reactors tapered with the time course, and at the steady state operation, the difference between two types of reactors on these two parameters was not obvious. Maximum specific methane or acids production rates, dehydrogenase activity and coenzyme F(420) content were continuously higher than those of the control reactors. After running 30 days, filamentous bacteria dominated in the reactors with pre-aeration, whereas the cocci were predominant species in the control reactors. It was suggested that the action of the biofilm is strongly dependent on the biofilm thickness or the biomass concentration in normal circumstances, but under adverse circumstances, such as organic or hydraulic loading shock, the characteristics and activity of the anaerobic granular sludge play key roles on the reactor performance. These results clearly indicated that pre-aeration of carrier favor to enhance the start-up and performance of AAFEB reactor. PMID- 15364090 TI - Extraction of oil from Jatropha curcas L. seed kernels by combination of ultrasonication and aqueous enzymatic oil extraction. AB - Use of ultrasonication as a pretreatment before aqueous oil extraction and aqueous enzymatic oil extraction was found to be useful in the case of extraction of oil from the seeds of Jatropha curcas L. The use of ultrasonication for 10 min at pH 9.0 followed by aqueous oil extraction gave a yield of 67%. However, the maximum yield of 74% was obtained by ultrasonication for 5 min followed by aqueous enzymatic oil extraction using an alkaline protease at pH 9.0. Use of ultrasonication also resulted in reducing the process time from 18 to 6 h. PMID- 15364091 TI - Reuse of waste materials as growing media for ornamental plants. AB - The use of different waste materials: pine bark, coconut fibre and sewage sludge as substrates in the production of ornamental plants was studied, with an special interest on the suitability of coconut fibre as growing substrate for conifer plants. The plant species tested were Pinus pinea, Cupressus arizonica and C. sempervirens and the substrate mixtures were: (1) pine bark, (2) pine bark with 15% of sewage sludge compost, (3) pine bark with 30% of sewage sludge compost, (4) coconut fibre, (5) coconut fibre with 15% of sewage sludge compost and (6) coconut fibre with 30% of sewage sludge compost. Substrates were physically and chemically well characterized, and 75-cm plants were grown on them for one year. Plant and substrate status were periodically tested along the experiment. As biosolid recycling is the main objective of the present work, the mixtures with 30% of composted sewage sludge will be the most convenient substrate to use. For C. sempervirens and C. arizonica, a mixture between pine bark or coconut fibre and 30% of biosolid compost in volume gave the best results, but the lower cost of the pine bark than the coconut fibre substrate indicated the use of the PB+30% CSS. For P. pinea the research of new combinations between waste products is recommended to attain better results. PMID- 15364092 TI - Data mining techniques for cancer detection using serum proteomic profiling. AB - OBJECTIVE: Pathological changes in an organ or tissue may be reflected in proteomic patterns in serum. It is possible that unique serum proteomic patterns could be used to discriminate cancer samples from non-cancer ones. Due to the complexity of proteomic profiling, a higher order analysis such as data mining is needed to uncover the differences in complex proteomic patterns. The objectives of this paper are (1) to briefly review the application of data mining techniques in proteomics for cancer detection/diagnosis; (2) to explore a novel analytic method with different feature selection methods; (3) to compare the results obtained on different datasets and that reported by Petricoin et al. in terms of detection performance and selected proteomic patterns. METHODS AND MATERIAL: Three serum SELDI MS data sets were used in this research to identify serum proteomic patterns that distinguish the serum of ovarian cancer cases from non cancer controls. A support vector machine-based method is applied in this study, in which statistical testing and genetic algorithm-based methods are used for feature selection respectively. Leave-one-out cross validation with receiver operating characteristic (ROC) curve is used for evaluation and comparison of cancer detection performance. RESULTS AND CONCLUSIONS: The results showed that (1) data mining techniques can be successfully applied to ovarian cancer detection with a reasonably high performance; (2) the classification using features selected by the genetic algorithm consistently outperformed those selected by statistical testing in terms of accuracy and robustness; (3) the discriminatory features (proteomic patterns) can be very different from one selection method to another. In other words, the pattern selection and its classification efficiency are highly classifier dependent. Therefore, when using data mining techniques, the discrimination of cancer from normal does not depend solely upon the identity and origination of cancer-related proteins. PMID- 15364093 TI - Finding optimal decision scores by evolutionary strategies. AB - OBJECTIVE: Severeness of illness is often rated by physicians at admission time. For this purpose, medical scores have been developed as 'objective' rating methods. When considering their classification performance, it is not assumed that such an expert-driven score is an optimal one. Our aim is to design an optimized data-driven score. In particular, we compare classical scores with a new data-driven score for abdominal septic shock patients. METHODS AND MATERIAL: Medical scores are used as ratings for different aspects of a patient's health status. The medical score indicates either a more critical or a healthier condition. For example, physicians rate organ conditions for different organs. We consider four different scores, SOFA, APACHE II, SAPS II, and MODS. Beyond the use of such classical scores, we propose an evolutionary strategy, that is suitable for score design, to find optimized data-driven scores. A database of 282 patients is used to optimize a new score for abdominal septic shock patients. Classification performance is compared by a ROC analysis. RESULTS: We give a general instruction for building optimized scores, i.e. we define individuals and operators for the evolutionary score design task. We apply this instruction to abdominal septic shock patient data. When compared to the SOFA score, it has similar classification performance, but it is more performant than APACHE II, SAPS II, and MODS. It can be used as a daily bedside score. CONCLUSIONS: We argue that evolutionary strategies should be used for optimizing purposes in the medical score design process. Using abdominal septic shock patient data, we show that evolutionary score design is a feasible and performant method that can complement or replace expert knowledge, provided that qualitative data is available. PMID- 15364094 TI - Measuring performance in health care: case-mix adjustment by boosted decision trees. AB - OBJECTIVE: The purpose of this paper is to investigate the suitability of boosted decision trees for the case-mix adjustment involved in comparing the performance of various health care entities. METHODS: First, we present logistic regression, decision trees, and boosted decision trees in a unified framework. Second, we study in detail their application for two common performance indicators, the mortality rate in intensive care and the rate of potentially avoidable hospital readmissions. RESULTS: For both examples the technique of boosting decision trees outperformed standard prognostic models, in particular linear logistic regression models, with regard to predictive power. On the other hand, boosting decision trees was computationally demanding and the resulting models were rather complex and needed additional tools for interpretation. CONCLUSION: Boosting decision trees represents a powerful tool for case-mix adjustment in health care performance measurement. Depending on the specific priorities set in each context, the gain in predictive power might compensate for the inconvenience in the use of boosted decision trees. PMID- 15364095 TI - Genetic design of feature spaces for pattern classifiers. AB - Functional piecewise approximation seeks data representation that is compact, highly simplified and meaningful. This study presents a genetic algorithm (GA) based approach for computing a piecewise polynomial representation of functions, with the focus being on piecewise linear approximation in an application of biomedical spectral data. The area of piecewise linear approximation has been researched in the past four decades approximately, and the method presented here is compared with another well-known approach. The expansion of this method to piecewise polynomial representation is shown to be straightforward. Finally, the application of this method as a feature extraction method for classification of a dataset of feature vectors, specifically biomedical spectra, is demonstrated. PMID- 15364096 TI - Detecting acute myocardial infarction in the 12-lead ECG using Hermite expansions and neural networks. AB - We use artificial neural networks (ANNs) to detect signs of acute myocardial infarction (AMI) in ECGs. The 12-lead ECG is decomposed into Hermite basis functions, and the resulting coefficients are used as inputs to the ANNs. Furthermore, we present a case-based method that qualitatively explains the operation of the ANNs, by showing regions of each ECG critical for ANN response. Key ingredients in this method are: (i) a cost function used to find local ECG perturbations leading to the largest possible change in ANN output and (ii) a minimization scheme for this cost function using mean field annealing. Our approach was tested on 2238 ECGs recorded at an emergency department. The obtained ROC areas for ANNs trained with the Hermite representation and standard ECG measurements were 83.4 and 84.3% (P=0.4), respectively. We believe that the proposed method has potential as a decision support system that can provide good advice for diagnosis, as well as providing the physician with insight into the reason underlying the advice. PMID- 15364097 TI - Medical privacy protection based on granular computing. AB - Based on granular computing methodology, we propose two criteria to quantitatively measure privacy invasion. The total cost criterion measures the effort needed for a data recipient to find private information. The average benefit criterion measures the benefit a data recipient obtains when he received the released data. These two criteria remedy the inadequacy of the deterministic privacy formulation proposed in Proceedings of Asia Pacific Medical Informatics Conference, 2000; Int J Med Inform 2003;71:17-23. Granular computing methodology provides a unified framework for these quantitative measurements and previous bin size and logical approaches. These two new criteria are implemented in a prototype system Cellsecu 2.0. Preliminary system performance evaluation is conducted and reviewed. PMID- 15364098 TI - Virulence regulation in Staphylococcus aureus: the need for in vivo analysis of virulence factor regulation. AB - Staphylococcus aureus is a pathogenic microorganism that is responsible for a wide variety of clinical infections. These infections can be relatively mild, but serious, life-threatening infections may result from the expression of staphylococcal virulence factors that are coordinated by virulence regulators. Much work has been done to characterize the actions of staphylococcal virulence regulators in broth culture. Recently, several laboratories showed that transcriptional analyses of virulence regulators in in vivo animal models or in human infection did not correlate with transcriptional analyses accomplished in vitro. In describing the differences between in vitro and in vivo transcription of staphylococcal virulence regulators, we hope to encourage investigators to study virulence regulators using infection models whenever possible. PMID- 15364099 TI - The solubilization and biological activities of Aspergillus beta-(1 --> 3)-D glucan. AB - We have recently demonstrated that the cell wall beta-glucan of Candida albicans could be solubilized by sodium hypochlorite, followed by dimethylsulfoxide extraction (NaClO-DMSO method). In this study, applying this method to Aspergillus spp., we prepared mycelial cell wall beta-glucan and examined its physical properties and immunotoxicological activity. The acetone-dried mycelia of Aspergillus spp. were oxidized by the NaClO-DMSO method. An analysis of (13)C NMR spectra revealed the preparations to be composed of alpha-(1 --> 3) and beta (1 --> 3)-D-glucan. Also, the proportion of alpha-(1 --> 3) and beta-(1 --> 3)-D glucan varied. Furthermore, a solubilized Aspergillus beta-glucan (ASBG) was prepared from OX-Asp by urea-autoclave treatment. ASBG showed limulus activity similar to Candida solubilized beta-glucan (CSBG), and there was little difference in the activity of ASBG between various Aspergillus spp. ASBG affected the production of IL-8 by human peripheral blood mononuclear cells (PBMC). ASBG should be useful for analyzing the clinical role of beta-glucan. PMID- 15364100 TI - Lethality in LPS-induced endotoxemia in C3H/HeCr mice is associated with prevalence of proinflammatory cytokines: lack of protective action of lactoferrin. AB - C3H/HeCr mice are more susceptible to infection compared with other strains. Lactoferrin (LF), a protein involved in innate defense, was shown to protect mice against lethal endotoxemia. In this investigation we attempt to explain the cause of increased susceptibility of C3H/HeCr mice to LPS and lack of protective LF action in these mice. We found that C3H/HeCr mice produced up to 5-fold more serum TNFalpha and 66% higher IFNgamma levels in response to i.v. LPS injection than the control, CBA strain. 24 h pretreatment of C3H/HeCr mice with LF did not cause inhibition of the LPS-induced TNFalpha serum levels, whereas in CBA mice LF significantly decreased TNFalpha level. IL-6 serum levels, in turn, were lowered in C3H/HeCr mice but elevated in CBA mice. That differential regulation of cytokine production by LF in C3H/HeCr mice paralleled a decreased survival after lethal LPS injection - 10% vs. 60% in control, PBS treated mice. In addition, determination of colony forming units (CFU) in livers and spleens after administration of 10(8) Escherichia coli revealed that pretreatment of CBA mice with LF caused a marked reduction of CFU in these organs, whereas in C3H/HeCr mice the changes were insignificant. These results indicate that the altered TNFalpha/IL-6 ratio in C3H/HeCr mice, as compared to control CBA mice, as well as the increased IFNgamma level, may be responsible for the increased susceptibility to endotoxemia in that substrain. We also suggest that an association exists between the LF protective effect against endotoxic sequelae and the insult induced systemic immune response. PMID- 15364101 TI - Patterns of cytokine induction by gram-positive and gram-negative probiotic bacteria. AB - Bacteria used in commercial probiotic preparations are most commonly gram positive lactic acid-producing species, although there are also some probiotic products which utilise gram-negative coliform bacteria. Characterising how the innate immune system responds to these bacteria in vitro may give an indication as to the likely immunomodulatory events that can be triggered following probiotic administration in vivo. Here, an established gram-positive probiotic (Lactobacillus casei Shirota) was compared against a novel gram-negative probiotic strain (Escherichia coli Nissle 1917) for its ability to induce cytokine production in a cell type representative of the innate immune system; in addition, responses were contrasted against those induced by an enteropathogenic coliform, E. coli 2282. We investigated the ability of these three bacterial strains to modulate production of interleukins-10, -12 and -18; tumour necrosis factor-alpha; interferon-alpha; and transforming growth factor-beta, via a series of in vitro culture experiments involving the murine monocyte/macrophage cell line J774A.1. All bacteria induced marked secretion of IL-12 and TNFalpha by cells, while only coliforms induced production of IL-10; there was minimal or no induction of IL-18 or TGFbeta. Activation of cells with recombinant gamma interferon promoted increased production of IL-12, but decreased production of IL 10, in response to the co-culture of coliform bacteria, indicating differential cytokine induction depending on the activation status of the target cell. In general, live bacteria stimulated higher levels of IL-10, IL-12 and TNFalpha secretion than heat-killed preparations, while only live coliforms induced IFNalpha. These findings are discussed in relation to the likely immunomodulatory effects of gram-positive and gram-negative bacteria on the innate immune system in vivo, with particular emphasis on the marked similarity in cytokine response patterns observed between probiotic versus pathogenic coliform bacteria. PMID- 15364102 TI - Effects of human lactoferrin on the cytoplasmic membrane of Candida albicans cells related with its candidacidal activity. AB - Human lactoferrin is an innate host defence protein with antimicrobial activity that exerts a candidacidal effect in a cation concentration-dependent manner. We investigated the ability of this cationic protein (with an isoelectric point of 8.7) to permeabilize the cytoplasmic membrane of Candida albicans cells. Despite minor K(+)-release in lactoferrin-treated C. albicans cells, the killing effect was not related to an extensive membrane permeabilization, as indicated by: (a) the non-release of macromolecular cytosolic constituents; (b) the non permeabilization for extracellular propidium iodide nor for intracellular accumulated calcein; and (c) the inability to disrupt the phospholipid bilayer of 8-aminonaphthalene-1,3,6, trisulfonic acid/p-xylene-bis-pyridiniumbromide-loaded liposomes. These results suggest that lactoferrin exerts its candidacidal effect through a mechanism different from membrane permeabilization described for other cationic peptides. PMID- 15364103 TI - The human immunodeficiency virus (HIV) protease inhibitor indinavir directly affects the opportunistic fungal pathogen Cryptococcus neoformans. AB - Highly active antiretroviral therapy (HAART), that includes human immunodeficiency virus (HIV) protease inhibitors (PIs), has been remarkably efficacious including against some opportunistic infections. In this report we investigated the effect(s) of the PI indinavir on protease activity by Cryptococcus neoformans, an opportunistic fungal pathogen responsible for recurrent meningoencephalitis in AIDS patients. Indinavir was also tested for potential effects on other parameters, such as fungal viability, growth ability and susceptibility to immune effector cells. It was found that indinavir impaired cryptococcal protease activity in a time- and dose-dependent fashion. The phenomenon was similarly detectable in ATCC/laboratory strains and clinical isolates. C. neoformans growth rate was also significantly reduced upon exposure to indinavir, while fungal viability was not affected and mitochondrial toxicity not detected. Furthermore, as assessed by an in vitro infection model, indinavir significantly and consistently augmented C. neoformans susceptibility to microglial cell-mediated phagocytosis and killing. Overall, by providing the first evidence that indinavir directly affects C. neoformans, these data add new in vitro insights on the wide-spectrum efficacy of PIs, further arguing for the clinical relevance of HAART against opportunistic infections in AIDS. PMID- 15364104 TI - Bacillus alcalophilus peptidoglycan induces IFN-alpha-mediated inhibition of vaccinia virus replication. AB - Bacterial products such as cell walls (CW) and peptidoglycan (PGN) are known to activate macrophages and NK cells during microbial infections. In this report, we demonstrated that whole CW and PGN of four Gram-positive bacteria are capable of enhancing the anti-poxviral activity of murine macrophage RAW 264.7 cells. Among the major Bacillus alcalophilus CW components, PGN contributes the most to antiviral activity and induces remarkably higher levels of IFN-alpha. Anti-IFN alpha/beta antibody, but not anti-IFN-gamma, anti-IFN-gamma receptor, or anti-IL 12, reversed the PGN-induced inhibition of vaccinia virus replication and reduced nitric oxide (NO) production. Our data thus suggest that PGN induce antiviral activity through IFN-alpha and to a lesser extent, through NO production. PMID- 15364105 TI - Epi p 1, an allergenic glycoprotein of Epicoccum purpurascens is a serine protease. AB - Epicoccum purpurascens (EP) is a ubiquitous saprophytic mould, the inhalant spores and mycelia of which are responsible for respiratory allergic disorders in 5-7% of population worldwide. The diagnosis/therapy of these disorders caused by fungi involves the use of standardized and purified fungal extracts. A 33.5 kDa glycoprotein, Epi p 1 released histamine from whole blood cells of EP allergic patients at a concentration of 50-ng protein. The high specific IgE values detected in EP hypersensitive sera indicated that Epi p 1 is capable of mediating type I hypersensitive reaction in predisposed individuals. It also showed protease activity by virtue of its dose dependent cleavage of serine protease specific synthetic substrate, N-benzoyl arginine ethyl ester hydrochloride (BAEE). The serine protease nature of Epi p 1 was confirmed by its N-terminal sequence (ADG/FIVAVELD/STY) homology to a subtilisin like serine protease. The protease activity of Epi p 1 may be responsible for making its way into the system of pre-disposed individuals through epithelial cell detachment and the histamine releasing ability by cross-linking of IgE antibodies on cell surface is the cause of its allergenic nature. PMID- 15364106 TI - Geographical distribution of antimicrobial resistance among Escherichia coli causing acute uncomplicated pyelonephritis in the United States. AB - The susceptibility to 12 antimicrobial agents of 165 Escherichia coli isolates from women with acute uncomplicated pyelonephritis of mild to moderate severity was analyzed by geographic region in the US. Ampicillin, trimethoprim, and trimethoprim/sulfamethoxazole resistance exhibited a descending prevalence gradient from west to east. Composite antimicrobial resistance phenotypes also exhibited significant regional differences, with a greater prevalence of most combined resistance profiles seen in the Pacific region of the US, but with significant north-south variation for combined ampicillin/sulfisoxazole resistance. These findings suggest geographical segregation of resistant clones and/or resistance elements among uropathogenic E. coli within the US, which is relevant both to clinical practice and to understanding the basis for the current epidemic of antimicrobial resistance in E. coli. PMID- 15364107 TI - Apoptosis of phagocytic cells induced by Candida albicans and production of IL 10. AB - Macrophages co-incubated with Candida albicans strain CR1 in vitro showed early signs of apoptosis, but evolved to necrosis after 2 h. In this study, we investigated whether strain CR1 caused apoptosis or necrosis of macrophages after its inoculation into mice peritoneal cavity, and whether this correlated with the secretion of IL-10. Peritoneal macrophages from mice that received an inoculum of C. albicans CR1 showed signs of apoptosis and necrosis from 30 min to 2 h afterwards, whereas heat-killed C. albicans did not cause those effects. IL-10 production was low during the first 6 h post-infection, when macrophages predominated in the peritoneal exudate, whereas its higher production after 24 h correlated with an increase of neutrophils in the exudate. Treatment of CR1 with pepstatin (an inhibitor of proteinases) prevented the process of apoptosis and significantly reduced IL-10 production, suggesting that the increased production of IL-10 was caused by processes occurring during the initial phase of infection, such as apoptosis, necrosis and uptake of death cells. PMID- 15364108 TI - Bacterial products increase expression of the human cathelicidin hCAP-18/LL-37 in cultured human sinus epithelial cells. AB - The respiratory epithelium plays a major role in the primary defense of the airways against infection. It has been demonstrated that bacterial products are involved in the induction of inflammatory reactions of the upper airways. Little is known about the effects of bacterial products on expression of the antimicrobial peptide hCAP-18/LL-37, the only human cathelicidin identified so far. The aim of this study was to investigate the effects of bacterial products from both gram-positive and gram-negative bacteria on the expression of hCAP 18/LL-37 by sinus epithelial cells using an air-exposed tissue culture model. Lipopolysaccharide and lipoteichoic acid both increased hCAP-18/LL-37 expression in cultured sinus epithelium as assessed by immunohistochemistry, where maximal stimulation occurred at 100 ng ml(-1) lipopolysaccharide or 10 microg ml(-1) lipoteichoic acid. The stimulatory effect of lipopolysaccharide and lipoteichoic acid was not restricted to expression of hCAP-18/LL-37, since also mucin expression and IL-8 release from cultured sinus epithelium cells were increased by lipopolysaccharide and lipoteichoic acid. This suggests that bacterial products may stimulate innate immunity in the upper airways. PMID- 15364109 TI - Cell mediated immune response elicited in mice after immunization with the P64k meningococcal protein: epitope mapping. AB - The P64k protein of Neisseria meningitidis has been reported as an immunological carrier for weak immunogens. This investigation was aimed at characterizing the T cell response produced in primed mice and at identifying T helper cell epitopes within this molecule. BALB/c mice subcutaneously immunized with the recombinant antigen provided inguinal lymph node cells (LNC) that proliferated in the presence of P64k in a dose-dependent manner. Proliferating cells secreted IL-4 while the concentration of IL-12 remained unaltered in the culture supernatant. By testing a panel of 59 overlapping synthetic peptides spanning the entire sequence of the antigen a T-cell determinant was localized. Prime-boost and lymphoproliferation experiments, conducted with highly purified synthetic peptides, confirmed that the segment including amino acids 470-485 comprises a T cell epitope within the P64k molecule. PMID- 15364110 TI - Vaccination with Leishmania soluble antigen and immunostimulatory oligodeoxynucleotides induces specific immunity and protection against Leishmania donovani infection. AB - In this report, we investigated the effect of ODN containing immunostimulatory CG motifs as adjuvant with soluble antigen (SA) from Leishmania donovani. BALB/c mice were vaccinated with the soluble antigen with or without CpG-ODN as adjuvant and then challenged with L. donovani metacyclic promastigotes. CpG-ODN alone resulted in partial protection against challenge with L. donovani. Immunization of mice with SA and CpG-ODN showed enhanced reduction in parasite load ( approximately 60%) when compared to SA ( approximately 40%) immunized mice. Immunization with SA by itself resulted in a mixed Th1/Th2 response whereas co administration of SA with CpG-ODN resulted in a strong Th1 promoting isotype as they together promoted production of immunoglobulin G2a. Leishmania-specific Th1 cytokine response was induced by co-administering CpG-ODN and SA as they together promoted production of IFN-gamma and IL-12. In the present study, we demonstrate that immunostimulatory phosphorothioate-modified ODN are promising immune enhancers for vaccination against visceral leishmaniaisis. PMID- 15364111 TI - Detection of bacterial DNA in blood samples from febrile patients: underestimated infection or emerging contamination? AB - We applied real-time broad-range polymerase chain reaction (PCR) to detect bacteraemia in blood from febrile patients. Interpretation of amplification results in relation to clinical data and blood culture outcome was complex, although the reproducibility of the PCR results was good. Sequencing analysis of the PCR products revealed the presence of Burkholderia species DNA while no Burkholderia species grew in culture. The source of this contamination was shown to be the commercial DNA isolation kit used in the automated MagNA Pure Isolation Robot. A high degree of suspicion is required when uncommon or unexpected pathogens are diagnosed by molecular methods as clinical consequences can be serious. PMID- 15364112 TI - Polymicrobial infections in children with diarrhoea in a rural area of Jordan. AB - Polymicrobial infections associated with diarrhoea are common in developing countries. Stool specimens were collected from 220 patient children and 100 controls. Potential pathogenic agents isolated from 143 (65%) children were identified by molecular and standard microbiological methods. Co-infections with two or more agents were detected in 50 (35%) cases. Escherichia coli, Shigella dysenteriae, Giardia and Entamoeba histolytica were found to be predominant. The etiologic agents could not be determined in 77 (35%) cases. The most significant risk factors were the age, the education level of the mother and the use of non chlorinated water. The high infection rate of diarrhoeal diseases is a strong indication that these pathogens circulate easily through the population. PMID- 15364113 TI - Retinoic acid treated HL60 cells express CEACAM1 (CD66a) and phagocytose Neisseria gonorrhoeae. AB - Neisseria gonorrhoeae (gonococci, GC) are phagocytosed by neutrophils through the interaction between opacity proteins (Opa) and the CEA (CD66) family of antigens. In order to study this interaction, we used the human myeloid leukemia HL60 cell line, which differentiates into granulocyte-like cells upon treatment with dimethylsulfoxide (DMSO) or retinoic acid (RA). We found that RA-, but not DMSO- or untreated-HL60 cells, can phagocytose OpaI-expressing gonococci as well as Escherichia coli. The interaction of OpaI E. coli with RA-treated HL60 cells was inhibited by antibodies against CEACAM1. Phagocytosis of OpaI E. coli was found to be a result of the expression of CEACAM1 in RA-treated HL60 cells. Our results indicate that the level of expression of CEACAM1 in HL60 cells can be regulated by treatment with RA in a differentiation-dependent manner, and that this is important for phagocytosis of OpaI-expressing gonococci or E. coli. PMID- 15364114 TI - Characterization of the human immune response to the UK anthrax vaccine. AB - The anthrax bipartite lethal toxin (protective antigen (PA) and lethal factor (LF))-specific antibody responses of humans receiving the UK licensed anthrax vaccine were determined. The PA-specific IgG response peaked two weeks post immunization and fell back to pre-boost levels by week 12. The heterogeneity of the host population modulated the extent of the PA-specific antibody response. Significantly lower levels of LF-specific antibodies were also detected. Vaccinated individuals recognized the same PA epitope as the protective mouse lethal toxin neutralizing monoclonal 2D3 suggesting that this may also be a target for human protection. PMID- 15364115 TI - Effects of acute diabetes on rat cutaneous wound healing. AB - INTRODUCTION:: Diabetes mellitus is a chronic hyperglycaemic disorder. Complicated metabolic mechanisms and increased incidence of infections are clinical hallmarks, mostly associated with its chronicity. There is little information about the early pathological processes in diabetes. The objective of our study was to evaluate the healing process during early phases of experimental diabetes on rat skin. METHODS:: Alloxan induced diabetic rats were used. Non injected animals were used as control. Punch byopsies on dorsal skin had histopathological evaluation of the healing areas made on days 1, 3 and 7 post surgery. RESULTS:: The results showed that: (1) in diabetics, the inflammation, the initial healing phase, has a slow beginning and tends to last longer; and (2) diabetic animals showed lower density of neutrophils in healing areas up to 3 days after surgery, and in addition, after day 3, when the neutrophils should leave the healing area, and be replaced by macrophages, compared to controls, diabetic animals showed higher numbers of neutrophils. PRINCIPAL CONCLUSION:: Although diabetes is a chronic progressive disease, acute diabetes can be associated to subclinical alterations, and responsible for deficiencies in defense cells and in repair tissue failures. PMID- 15364116 TI - Vascular endothelial growth factor - basic science and its clinical implications. AB - Vascular endothelial growth factor (VEGF) is the most important signaling molecule involved in the regulation of the formation of new vessels. Results of recent studies have provided new insights into the molecular mechanisms of the VEGF signaling pathways. VEGF local or systemic application represents a new approach in the therapy of ischemic diseases, especially of the coronary artery disease. Inhibition of the VEGF action on various levels is, on the other hand, assumed to be a promising therapeutic concept against cancer. Moreover, VEGF has been recently shown to be associated with some other physiological and pathophysiological processes. In this article we summarize the latest results of VEGF related studies and present the concluding theoretical resource for further research on the role of VEGF in understanding of pathophysiology of diseases and in therapeutic interventions in clinical biomedicine. PMID- 15364117 TI - Direct nitric oxide release from nipradilol in human coronary arterial smooth muscle cells observed with fluorescent NO probe and NO-electrode. AB - BACKGROUND:: Nipradilol (3,4-dihydro-8-[2-hydroxy-3-isopropyl-amino]propoxy-3 nitroxy-2-H-1-benzopyran), a potent non-selective beta-adrenoceptor antagonist, has been shown to increase NO production. The mechanisms are up-regulation of nitric oxide synthase (NOS) and direct release of NO from nipradilol. The process of direct NO release from nipradilol requires a reductase, such as glutathione S transferase (GST) in some cells but non-enzymatic NO release was reported in pig coronary arteries. Direct NO release from nipradilol in human coronary arteries has not been examined yet, though this information is of importance. PURPOSE:: To demonstrate direct NO release from nipradilol in human coronary arterial smooth muscle cells (HCASMC) by using a fluorescent NO probe (DAF-2) and an NO electrode. METHODS AND RESULTS:: HCASMC were loaded with DAF-2 and images of fluorescence (515nm) were obtained under excitation at 488nm through an intensified CCD with an inverted phase-contrast microscope. Concomitantly, NO was measured using an NO-electrode (0.2mm o.d.; 501, Inter Medical Co. Ltd., Nagoya, Japan) after addition of various concentrations of nipradilol (1, 5 or 10microM) with or without ethacrynic acid (GST inhibitor). The cells showed no fluorescence at baseline, but intense fluorescence appeared at 30min after addition of 10microM nipradilol. The intensities of fluorescence at 30min in the control, nipradilol and nipradilol with ethacrynic acid groups were 98 +/- 6, 163 +/- 10 and 128 +/- 6% of the baseline level, respectively. Ethacrynic acid itself did not affect the fluorescence. Continuous measurements of NO by the electrode showed the NO generation peaked at about 30min, remained at the same level till about 45min and then gradually declined. Nipradilol did not produce NO at all in the absence of cells. The dose-dependency study of NO release from nipradilol showed 45 +/- 12, 72 +/- 24 and 157 +/- 23nM, respectively, at 1, 5 and 10microM nipradilol. All experiments were performed under conditions where endogenous formation of NO was inhibited by an NOS inhibitor (10(-4)M N(G)-monomethyl-l arginine (l-NMMA)). CONCLUSION:: Nipradilol can release NO in the presence of human coronary arterial smooth muscle cells and the denitration reaction catalyzed by a reductase such as glutathione S-transferase contributes substantially to NO release from nipradilol. PMID- 15364118 TI - Delta-sleep inducing peptide and neuronal activity after glutamate microiontophoresis: the role of NMDA-receptors. AB - The influence of microiontophoretic application of delta-sleep inducing peptide (DSIP) on a neuronal activity of sensorimotor brain cortex, dorsal hippocampus, ventral anterior thalamic nucleus and lateral hypothalamus was studied under the effects of glutamate and MK-801, a N-methyl-d-aspartate non-competitive antagonist, on male Wistar rats. DSIP microiontophoresis predominantly activated neurons of various brain regions, in particular, dorsal hippocampus and ventral anterior thalamic nucleus. A preliminary DSIP microiontophoresis prevented the augmentation of a neuronal activity in the studied structures under glutamate microiontophoresis. After MK-801 microiontophoresis the number of neurons activated by the DSIP significantly decreased. It is suggested that DSIP effects on the neuronal activity in the sensorimotor brain cortex, dorsal hippocampus, ventral anterior thalamic nucleus and lateral hypothalamus might be mediated by the NMDA-receptors. PMID- 15364119 TI - Spiral artery associated restricted growth (SPAARG): a computer model of pathophysiology resulting from low intervillous pressure having fetal programming implications. AB - Failure of adequate trophoblastic conversion of maternal spiral arteries is associated with intrauterine growth restriction (IUGR). In addition to poor oxygen delivery, raised spiral artery resistance reduces placental intervillous pressure. An iterative type computer model was formed by linking an existing model of the fetus and a new nine cotyledon placental model. Simulation of compression cuffing of the spiral arteries to progressively restrict uteroplacental flow was performed, while observing various fetal and placental variables. Water moved to the fetus in the cotyledonary core villi, and to the mother in the outer villous layers. While the fetus could match villous capillary pressure to changes in intervillous pressure, net transplacental water movement was minimal, but when spiral artery resistance was increased sufficiently to cause mean intervillous pressure to fall below that which the fetus could match, a net flow to the mother appeared. That continued until the resulting fetal blood hemoconcentration produced a sufficient increase in colloid osmotic pressure to restrict further loss. All cells within the fetal-placental unit are then required to operate in an abnormal ionic environment, which may significantly affect systems such as the renin-angiotensin set-point, with implications for post-natal homeostasis such as control of adult blood pressure. Furthermore, in vivo, cells of the feto-placental unit respond to the increased intravascular osmotic pressure by production of intracellular osmolytes in order to match intracellular and vascular/interstitial osmotic pressures. This may explain the observed effects on postnatal water balance in growth restricted infants and could also provide a possible mechanism for the association of the systemic maternal complications associated with impaired placentation and reduced intervillus flow. PMID- 15364120 TI - Calcification in coronary artery disease can be reversed by EDTA-tetracycline long-term chemotherapy. AB - Atherosclerosis is a complex process with multiple mechanisms and factors contributing to its initiation and progression. Detection and quantification of coronary artery calcium (CAC) scores with electron beam tomography has been shown to correlate with obstructive and nonobstructive coronary artery disease (CAD). Pathogen-triggered calcification could play a role in CAD. Recent reports suggest that infectious blood nanobacteria (NB) emerge to be such a trigger. So far, minimal or no reversal of atherosclerosis has been claimed by therapies with iv ethylenediaminetetraacetic acid disodium salt (EDTA), antibiotics, or other regimens, and therapies for atherosclerosis remain non-curative. We have now combined EDTA with antibiotic tetracycline (comET), an in vitro proven nanobacteriocidal treatment, and tested comET therapy in patients with documented CAD. Three hypotheses were probed: (1) Are NB present in patients with CAD?; (2) Does treatment with comET affect blood NB antigen and serology?; (3) Does a comET decrease CAC scores? One hundred patients with stable CAD and positive CAC scores were enrolled into a 4 month study of comET therapy. ComET therapy is composed of (1) Nutraceutical Powder (Vitamin C, Vitamin B6, Niacin, Folic Acid, Selenium, EDTA, l-Arginine, l-Lysine, l-Ornithine, Bromelain, Trypsin, CoQ10, Grapeseed Extract, Hawthorn Berry, Papain) 5cm(3) taken orally every evening; (2) Tetracycline HCl 500mg taken orally every evening; (3) EDTA 1500mg taken in a rectal suppository base every evening. CAC scoring was repeated at 4 months and serum samples were analyzed for NB antigen and serology at baseline, 2 and 4 months. Complete blood count, metabolic panel, liver function, C-reactive protein (hs-CRP) and lipids were analyzed at baseline and 4 months. Seventy-seven patients completed the study and all patients were positive for NB serology, antigen or both. Responders (n = 44; 57%) had significant decreases in total CAC scores (P = 0.001), the average decrease being 14%. Non-responders (n = 33; 44%) had no change or had increases in CAC scores. Angina was decreased or ablated in 16 of 19 patients (84%). Lipid profiles improved to non-atherogenic direction significantly (P = 0.001), a remarkable finding in a patient group where 86% were on continuous statin medication already before the trial. No adverse physiologic effects were seen in renal, hepatic, or hematopoetic systems. In conclusion, CAC scores decreased during ComET therapy trial in most CAD patients inferring regression of calcified coronary artery plaque volume. The patients tolerated the therapy well and their angina and lipid profiles improved. Further treatment trials for long term therapy with matched controls are warranted. PMID- 15364121 TI - Method improvement: released cecal ligation-septic shock model. AB - A new released cecal ligation (RCL) technique of septic shock induction in rats is presented. It is relatively not complicated, fast, not expensive, and it mimics the human pathology. Presented procedure seems to be a useful alternative for the cecal ligation alone or cecal ligation and puncture to induce experimental septic shock in small laboratory animals. PMID- 15364122 TI - Administration of high dose intravitreal melatonin degenerates retinal cells in guinea pigs. AB - The purpose of this study was to investigate the influence of intravitreally injected different doses of melatonin on retinal morphology. The right eyes of 35 male albino guinea pigs were used. The animals were classified randomly into five groups in equal numbers. First group was used as control and received intravitreal injection of placebo. Groups 2, 3, 4 and 5 received intravitreally injections of melatonin at 50, 100, 150 and 200microg/body weight (BW) each, respectively. The animals were sacrificed 15 days after the injections. The eyes were enucleated and processed for light microscopic evaluation. Intravitreal injection of melatonin at doses ranging from 50 to 150microg did not induce morphological changes, although a higher thickness of the inner plexiform layer (IPL) was found in Group 5 compared to other groups (p < 0.05). The mean retinal ganglion cell (RGC) counts were found to be lower in Group 5 compared to other groups (p < 0.05). Our findings indicate that intravitreal injection of melatonin at doses ranging from 50 to 150microg/BW does not induce morphological changes. The dose of 200microg/BW produced significant damage including retinal ganglion cell loss and formation of retinal edema. PMID- 15364123 TI - Atherogenesis and vascular calcification in mice expressing the human LPA gene. AB - Background: Lp(a) lipoprotein (Lp(a)) contains polymorphic glycoprotein, apolipoprotein(a) (apo(a)) and low density lipoprotein (LDL). The extensive homology between apo(a) and plasminogen is believed to contribute to the pathogenicity of apo(a), but the precise mechanisms by which Lp(a) participates in atherogenesis is still unknown. We used LPA-yeast artificial chromosome (LPA YAC) transgenic mice with or without the human APOB (hAPOB) gene to study pathogenicity of apo(a)/Lp(a) and illucidate its role in regulation of serum lipid levels. Methods: Middle-aged (1-year-old) mice were fed a control (AIN-76), a high-cholesterol (HC) or a high-cholesterol/high-fat (HCHF) diet for 7 weeks. For the study of serum total apo(a) and lipid levels, mice were sampled prior to the experiment, at 2 weeks and at 7 weeks when the animals were sacrificed. Hearts with ascending aorta were fixed in formalin, embedded in gelatine and prepared for sections on a cryostat. Livers were washed in ice cold saline and submerged in RNAlater trade mark buffer and stored at -70 degrees C until mRNA analysis. Results: Wild type mice fed the control diet did not develop aortic lesions. Presence of the LPA gene was sufficient to induce development of aortic lesions, but neither coexpression of the hAPOB gene nor feeding the HC diet or the HCHF diet augmented the development of aortic lesions in LPA-YAC transgenic mice. On the control diet transgenic females had larger aortic lesion size than transgenic males. Furthermore, aortic lesions in transgenic females were associated with calcification more often than in transgenic males. Serum total cholesterol levels were higher both in wild type and LPA-YAC transgenic males than in females mainly because of higher serum high-density lipoprotein cholesterol levels. HC and HCHF feeding had more pronounced effect on total cholesterol levels in LPA-YAC/hAPOB transgenic mice than in either wild type or LPA-YAC transgenic mice, due to increased low density lipoprotein cholesterol levels. Furthermore, these diets reduced serum total apo(a) levels in both transgenic mouse lines. Conclusion: Expression of the human LPA gene in mice is sufficient to trigger development of aortic lesions. Similar frequency of calcified lesions in LPA-YAC transgenic mice with or without hAPOB gene may suggest that apo(a) is the part of the Lp(a) molecule that causes aortic calcification. The basis for reduced serum total apo(a) level in response to cholesterol feeding is not clear, but interplay between LPA and factors involved in cholesterol or bile acid homeostasis is worth of future studies. PMID- 15364124 TI - Direct inhibition by a statin of TNFalpha-induced leukocyte recruitment in rat pial venules - in vivo confocal microscopic study. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) have been shown to block leukocyte-endothelial interaction independently of their cholesterol-lowering properties. The effects of statins are generally attributed to a decrease in mevalonate caused by inhibition of HMG-CoA reductase, which results in an increase of nitric oxide (NO). However, a recent in vitro study demonstrated a novel effect which depended on the lipophilicity of statin and appeared to be unrelated to HMG-CoA reductase inhibition. The purpose of this study is to investigate whether the proposed mechanism actually operates in vivo. We examined the effects of simvastatin (lipophilic) and pravastatin (hydrophilic) on leukocyte behavior in a tumor necrosis factor alpha (TNFalpha)-induced leukocyte recruitment model. Leukocyte adhesion and rolling were examined in pial venules of rat brain by using confocal laser scanning microscopy after labeling leukocytes with rhodamine 6G. Experiments were conducted 4h after TNFalpha injection (0.5microg) in six groups: control, TNFalpha alone, TNFalpha + vehicle of simvastatin, TNFalpha + simvastatin (20mg/kg, 2ml/kg), TNFalpha + vehicle of pravastatin, and TNFalpha + pravastatin (40mg/kg, 2ml/kg). Statins and vehicles were injected subcutaneously for 3 days. TNFalpha caused a marked increase in rolling and adhered leukocytes. The number of adhered leukocytes in the simvastatin group was significantly less than in the vehicle group (276 +/- 38 cells/mm(2) versus 1155 +/- 89 cells/mm(2), P < 0.01), whereas pravastatin had little effect. Both simvastatin and pravastatin showed a tendency to decrease the number of rolling leukocytes, but there were no significant differences among TNFalpha-treated groups. Up-regulation of endothelial nitric oxide synthase (eNOS) mRNA or increased expression of P-selectin or intercellular adhesion molecule-1 (ICAM-1) was not observed, and therefore cannot account for the simvastatin-induced reduction of adhered leukocytes. Markedly different effect on leukocyte adhesion between simvastatin and pravastatin under comparable level of HMG-CoA reductase inhibitor was demonstrated in in vivo as was shown in in vitro study. PMID- 15364125 TI - Garlic induced alteration in liver mineral concentrations in corn oil and olive oil fed rats. AB - This study was undertaken to compare the effects of 2% garlic (G) on liver mineral concentrations in rats fed with corn oil (C) or olive oil (O). Male Sprague-Dawley rats (n = 4 per group) were fed AIN76A semi-purified diets containing either 5% corn oil or 5% olive oil replacing corn oil with or without 2% garlic for 21 days. The analysis of minerals calcium (Ca), phosphorous (P), potassium (K), magnesium (Mg), manganese (Mn), sodium (Na), Sulfur (S), copper (Cu), iron (Fe) and zinc (Zn) of liver were conducted by inductively coupled plasma (ICP) spectroscopic method. In the C fed rats addition of 2% garlic produced significant increase in liver Ca, P, Mg, S, Mn, Cu, and Zn. There was no significant change in Fe, Na and K. In contrast, in olive oil fed rats, 2% garlic diets did not affect Ca, Mg, S, Mn, Na and K concentrations. However, there was significant decrease in liver Cu and Zn concentrations. Also, in O fed control rats liver, Ca, P, Mg, S, Fe, Mn, Cu, and Zn concentrations were significantly increased as compared to C fed control rats. In conclusion, this study describes the interactions between dietary oils and garlic on liver mineral concentrations in rats. PMID- 15364126 TI - Abstracts of the 4th International Conference on Early Psychosis. Vancouver, Canada, September 28-October 1, 2004. PMID- 15364127 TI - DNA repair gene XPD polymorphism and lung cancer risk: a meta-analysis. AB - Interindividual variation in lung cancer susceptibility may be modulated in part through genetic polymorphisms in the DNA repair genes, especially the genes involved in the nucleotide excision repair (NER) pathway. The xeroderma pigmentosum complementary group D (XPD) is one of the NER genes, and two of the XPD polymorphisms 751A --> C and 312G --> A have been extensively studied in the association with lung cancer, although published studies have been inconclusive. To clarify the impact of XPD polymorphisms on lung cancer risk, we performed a meta-analysis of the published data from nine (10 comparisons) individual case control studies of 3725 lung cancer cases and 4152 controls. The results showed that individuals with the XPD 751CC genotype had a 21% (odds ratio (OR)= 1.21, 95% confidence interval (CI) = 1.02-1.43) increased risk of lung cancer compared with individuals with the 751AA genotype without any between-study heterogeneity (P = 0.26). There was also a significant association in the recessive model of 751 C allele by comparing the CC with AC + AA genotypes (OR = 1.19, 95% CI = 1.02 1.40). The results also showed a significantly increased risk of lung cancer associated with the 312AA homozygous genotype compared with the GG genotype and the 312 A allele in the recessive model (compared with GA + AA genotypes) (OR = 1.27, 95% CI = 1.04-1.56 and OR = 1.32, 95% CI = 1.09-1.59, respectively). These results support the hypothesis that both the XPD 751 C and 312 A are risk alleles and individuals with the XPD 751 CC and 312 AA genotypes are at higher risk of developing lung cancer. Large multi-center studies with precise design, and stratified/adjusted analyses of the gene-gene (haplotypes) and gene-environment interactions are needed. PMID- 15364128 TI - Human small cell lung cancer cells express functional VEGF receptors, VEGFR-2 and VEGFR-3. AB - Studies have suggested that the vascular endothelial growth factors (VEGFs)/VEGF receptors (VEGF-Rs) system plays an important role in tumour growth and metastasis. We conducted the present study to clarify whether small cell lung cancer (SCLC) cells express functional VEGF-Rs and VEGFs, and their biological significance in the SCLC progression. We examined expression of VEGF and VEGF-C, and their receptors, VEGFR-2 and VEGFR-3, in five SCLC cell lines, NCI-H82, H209, H510, H526 and H660, by Western blotting. We evaluated whether hypoxic conditions up-regulate these protein expressions. We also examined whether VEGF addition and VEGF-D addition cause phosphorylation of the mitogen-activated protein kinase (MAPK) as well as VEGFR-2 and VEGFR-3. Further, we investigated whether VEGF addition and VEGF-D addition induced the proliferation and migration of the SCLC cells. VEGF, VEGF-C, VEGFR-2 and VEGFR-3 were detectable by Western blotting in all five SCLC cell lines,. The VEGF-Rs and VEGFs expression levels were increased by an incubation under hypoxic conditions in NCI-H82. VEGF addition and VEGF-D addition caused phosphorylation of MAPK as well as the VEGF-Rs themselves, and induced proliferation and migration of the SCLC cells. These results suggested potential of VEGF signal-pathway inhibitors as anti-cancer agents in SCLC treatment disturbing growth and migration of the cancer cells. PMID- 15364129 TI - Expression of LUN gene that encodes a novel RING finger protein is correlated with development and progression of non-small cell lung cancer. AB - LUN is a novel RING finger protein that is highly expressed in the lung and might be a transcriptional regulator of E-cadherin [J. Biol. Chem. 276 (2001) 14004]. It might be possible that LUN plays important roles in the development and progression of lung cancer through regulating expression of E-cadherin, but no clinical study on LUN expression has been reported. In the present study, we quantitatively examined gene expression of the LUN in surgical specimens resected from non-small cell lung cancer (NSCLC) patients. In normal lung tissues, the LUN gene expression was down-regulated in smokers (the mean LUN/GAPDH ratios, 0.222 for non-smokers and 0.144 for smokers; P = 0.030). In addition, the mean LUN/GAPDH ratio in lung cancer tissues was significantly lower than that in normal lung tissues (0.072 versus 0.162; P < 0.001). In addition, the LUN gene expression was slightly down-regulated along with progression of primary tumors, and strongly down-regulated along with nodal metastases (the mean LUN/GAPDH ratios, 0.091 for pN0, 0.073 for pN1, and 0.034 for pN2 diseases; P = 0.001). These results suggested that LUN might play important roles in inhibition of nodal metastases as well as in suppression of smoking-related oncogenesis in NSCLC. PMID- 15364130 TI - Prognostic significance of gelsolin expression level and variability in non-small cell lung cancer. AB - BACKGROUND: Gelsolin is an actin-binding protein that mediates cellular motility and maintains the integrity of cytoskeletal structure. Diminished expression of gelsolin has been observed in human cancer cell lines and tumors. Studies of the prognostic effect of gelsolin expression (GE) in non-small cell lung cancer (NSCLC) are rare and results are inconsistent to date. The present study used immunohistochemistry to evaluate the prognostic effect of gelsolin expression in 155 patients with resectable NSCLC. METHODS: Detection of gelsolin in tumor cells was performed by immunohistochemistry, and two approaches to classification were used to describe expression: expression level (negative, reduced or high) and expression uniformity (uniform or variable). Expression level was determined by a weighted index of intensity of staining (i.e., overall tendency) in the specimen. Expression uniformity was based on the presence or absence of variability in immunostaining within the tumor section. Chi-square test, student t-test, Cox proportional hazards regression and Kaplan-Meier survival analysis were used in data analyses. RESULTS: After controlling for covariates, high level gelsolin expression was significantly associated with poor survival compared with negative gelsolin expression in NSCLC, and this adverse prognostic effect was specific to patients with stage II tumors and for patients with squamous cell carcinomas. Similarly, variable gelsolin expression was significantly associated with poor survival compared with uniform gelsolin expression and this adverse prognostic effect was also specific to patients with stage II tumors and for patients with squamous cell carcinomas. CONCLUSION: High level gelsolin expression and variable gelsolin expression are adverse prognostic factors for NSCLC in this study, which might manifest the high motility and heterogeneity of tumor cells, two distinguishing characteristics for tumors with potentially enhanced invasive and dissemination capabilities. PMID- 15364131 TI - Usefulness of ultrathin bronchoscopy in diagnosis of lung cancer. AB - Although ultrathin bronchoscopes are suggested to have comparable abilities to conventional bronchoscopes in diagnosing peripheral lung lesions, how to introduce ultrathin bronchoscopes into bronchoscopic examination is still to be determined. In our first study, 35 patients with peripheral lung lesions underwent ultrathin followed by conventional bronchoscopy to compare their diagnostic abilities. The diagnostic rate was 54.3% in conventional bronchoscopy alone, 60.0% in ultrathin bronchoscopy alone, and 62.8% in the combination of the two. In the next study, we introduced a rapid cytology test of the material obtained in conventional bronchoscopy. When malignant cells were not detected in the material by the rapid cytology, ultrathin bronchoscopy was immediately conducted. Thirty-two patients with negative rapid cytology were enrolled in this study. Ultrathin bronchoscopy resulted in diagnostic materials in 59.3% of these cases. Ultrathin bronchoscopes showed better access to the lesions than a brush or a curette introduced through conventional bronchoscopes. We conclude that ultrathin bronchoscopes have a comparable ability to conventional ones in diagnosing peripheral lung cancer even when used alone, and become a good complement to conventional ones by introducing the rapid cytology test. PMID- 15364132 TI - In vivo identification of sentinel lymph nodes for clinical stage I non-small cell lung cancer for abbreviation of mediastinal lymph node dissection. AB - BACKGROUND: We previously reported that an identification of sentinel lymph node (SN) with a techenetium-99m (99mTc) tin colloid by ex vivo counting, i.e. the radio-activity of dissected lymph nodes, was a reliable method of establishing the first site of nodal metastasis in non-small cell lung cancer [J. Thorac. Cardiovasc. Surg. 124(2002)486]. However, for SN navigation surgery, SN should be identified before lymph node dissection (in vivo) but not after that (ex vivo). In order to reduce mediastinal lymph node dissection for clinical stage I non small cell lung cancer (NSCLC) by SN navigation surgery, the SN identifications for hilar lymph nodes by ex vivo counting, and for mediastinal lymph nodes by in vivo, were evaluated. METHODS: Intra-operative SN identification using 99mTc tin colloid was conducted on 104 patients with clinical stage I NSCLC who had had major lung resections with mediastinal lymph node dissections. The hilar SNs were identified by ex vivo counting (after lung resection) and the mediastinal SNs were identified by in vivo counting (before lymph node dissection). To evaluate the accuracy of mediastinal SN identification by in vivo counting, it was compared with the data by ex vivo counting. RESULTS: SNs were identified in 84 patients (81%). SNs were identified at the hilum by ex vivo counting in 78 patients (93%) and at the mediastinum by in vivo counting in 40 patients (48%). While 15 patients had lymph node metastases, i.e. N1 in six and N2 in nine, the SNs could be found to have metastases during operation in 13 of the 15 patients (87%). The in vivo counting of the mediastinum missed out the mediastinal SNs identified by ex vivo counting in four of the 84 patients (5%). CONCLUSION: If the hilar SNs identified by ex vivo counting and the mediastinal SNs identified by in vivo counting had no metastases, then mediastinal lymph node dissection could be abbreviated for patients with clinical stage I NSCLC. PMID- 15364133 TI - Increasing social and psychological information conveyed to General Practitioners after bad news consultations by use of separate letters from the Lung Cancer Nurse Specialist. AB - Multidisciplinary teams (MDT) now review all cases of lung cancer. These teams include a Lung Cancer Nurse Specialist (LCNS). These Nurses help support the patient and should facilitate communication and liaise with other services. The LCNS is present when the diagnosis is given to the patient but also usually spends time afterwards with the patient and their family. We postulated that a separate letter from the LCNS to the General Practitioner (GP) after the consultations would convey extra information to the GP. In 58 new lung cancer patients reviewed in the clinic, the LCNS and Physician independently wrote separate letters after the consultation in which the diagnosis of lung cancer was given. The GPs were asked by questionnaire about the usefulness of the letter from the LCNS. This letter was considered by the GP to provide extra information in: (i) 69% concerning the patients reaction to the diagnosis; (ii) 85% concerning who attended the clinic with the patient; (iii) 85% about what referrals were made to community services; (iv) 86% about who the patient was living with; (v) 81% about who the patients carers were; (vi) 81% information on the patients condition; (vii) 70% concerning the information given to patients about benefits. Ninety-seven percent of the GPs found the LCNS letter useful or very useful and 92% of the GPs thought that the information in the letter would be useful or very useful when they next saw the patient. Separate and independent letters from the LCNS after "bad news" consultation in lung cancer provides added useful information for GPs. Ninety-one percent of the GPs wanted the letters from the LCNS to continue. PMID- 15364134 TI - An overview of 48 elderly-specific clinical trials of systemic chemotherapy for advanced non-small cell lung cancer. AB - PURPOSE: The aim of the present study was to identify elderly-specific clinical trials for advanced non-small cell lung cancer (NSCLC) and to clarify the study design and patient characteristics entered of each of these trials. METHODS: We used the MEDLINE database to select prospective clinical trials evaluating the efficacy of chemotherapy in elderly patients with advanced NSCLC. RESULTS: Our literature search yielded 48 prospective clinical trials between 1990 and 2003, involving a total of 2648 elderly patients with advanced NSCLC. The median number of patients treated per trial was 36. In 23 (48%) of the 48 trials, only the abstract was available. In 44 trials (92%), elderly patients were defined using their calendar age, and the age of 70 years was the most frequently used lower limit for inclusion. Vinorelbine was the most widely studied chemotherapy agent in elderly patients. CONCLUSIONS: Our review revealed that (i) the definition of "elderly" varied from trial to trial, and elderly patients were simply defined using calendar age in the clinical trials; (ii) the quality of elderly-specific trials were generally poor, mainly because of their small sample size. PMID- 15364135 TI - Vascular endothelial growth factor, p53, Rb, Bcl-2 expression and response to chemotherapy in advanced non-small cell lung cancer. AB - Vascular endothelial growth factor (VEGF) increases microvascular permeability and stimulates endothelial cell growth. p53 Overexpression has been associated with resistance to cisplatin-based chemotherapy in patients (pts) with NSCLC. The aim of this study was to evaluate the predictive role of VEGF for chemotherapy response, its relationship with p53, Rb, Bcl-2 and hemoglobin levels and its impact on overall survival in pts with advanced NSCLC. Bronchial or fine-needle biopsy specimens from 85 pts with NSCLC obtained before chemotherapy were analyzed using an immunohistochemical method for VEGF, p53, Rb and Bcl-2. There were 73 males and 12 females with a median age of 62.6 years. The majority of pts (48%) had squamous cell histology. Ten pts had stage IIIA, 25 stage IIIB and 50 stage IV. Thirty six (43%) pts had positive immunostaining for VEGF, 37 (44%) had positive p53, 53 (62%) had negative Rb and 4 (5%) had positive Bcl-2. VEGF was negatively correlated with Rb (r(s) = 0.26; P = 0.015), positively with Bcl-2 (r(s) = 0.22; P = 0.42), whereas no statistically significant correlation with p53, age, stage and histological type was found. In a logistic regression model, adjusting for treatment, VEGF expression was not associated with chemotherapy response (odds ratio (OR) = 1.01; P = 0.085 ), unlike p53 positivity and Rb negativity ( OR = 4.0, P = 0.005; OR = 2.6, P = 0.016, respectively). A statistically significant higher VEGF expression was detected in the subgroups defined, using as cut-off value Hb median level (13.3g/dl) (chi-square = 5.00; ; one d.f.; P = 0.025). At a median follow-up time of 8.4 years, 2-year survival was 21%. After adjustment for stage and chemotherapy treatment, VEGF expression was not associated with a better overall survival (OR = 1.06; P = 0.80), unlike Bcl-2 positivity showed a statistically significant effect (OR = 0.28; p = 0.02). Our results suggest that VEGF is weakly correlated with regulators of apoptosis and has not been shown to be an independent predictive factor for resistance to cisplatin-based chemotherapy and prognostic for survival. PMID- 15364136 TI - Concurrent versus sequential chemoradiotherapy with cisplatin and vinorelbine in locally advanced non-small cell lung cancer: a randomized study. AB - PURPOSE: The superiority of chemoradiotherapy (CRT) over radiation alone in locally advanced non-small cell lung cancer (NSCLC) has been proven, but the relative merits of a concurrent schedule versus their sequential administration are less clear. This study compared the safety and efficacy of concurrent and sequential CRT, with chemotherapy (CT) consisting of a cisplatin and vinorelbine regimen, in patients with locally advanced NSCLC. PATIENTS AND METHODS: One hundred and two previously untreated patients (aged 42-75 years) with locally advanced, stage IIIA (n = 15) or stage IIIB (n = 87) NSCLC were entered into the study. The CT schedule consisted of up to four cycles of cisplatin 80 mg/m(2) on day 1, and vinorelbine 25 mg/m(2) at the first and fourth cycles (12.5 mg/m(2) during the 2nd/3rd cycles) on days 1, 8, 15 of a 28-day cycle. Radiotherapy (RT) was prescribed at a dose of 60 Gy/30 fractions, given as five fractions per week for 6 weeks. In the concurrent arm (arm A), RT was started on day 4 of cycle 2; whilst in the sequential arm (arm B), RT started within 2 weeks after completion of CT. Fifty-two patients were randomized to concurrent treatment and 50 to the sequential schedule. RESULTS: Overall survival was significantly longer in arm A (median survival 16.6 months) versus arm B (median survival 12.9 months) (P = 0.023 by means of log-rank test; hazard ratio HR = 0.61, 95% CI of HR (0.39 0.93)), and time to progression (TTP) was also significantly longer in arm A (median time to progression 11.9 months) versus arm B (median time to progression 8.5 months) (P = 0.024 by means of log-rank test; HR = 0.62, 95% CI of HR (0.38 0.93)). Ninety-eight patients were evaluable for response and 101 for toxicity. The overall response rate was significantly higher in arm A, 80% (with 21% complete response (CR)) compared with 47% (with 17% CR) in arm B (P = 0.001 by means of chi(2)-test). WHO grade 3 or 4 toxicity was more frequent in arm A than in arm B, with a significantly greater incidence of leucopenia (53% versus 19%, P = 0.009 by means of chi(2) test) and nausea/vomiting (39% versus 15%, P = 0.044 by means of chi(2) test). There were no treatment related deaths. CONCLUSION: In this study population, concurrent CRT demonstrated significant benefit in terms of response rate, overall survival and time to progression over sequential CRT. The concurrent CRT schedule was associated with higher toxicity; however, the adverse event profile was acceptable in both arms. PMID- 15364137 TI - Sequential chemotherapy with paclitaxel plus cisplatin, followed by vinorelbine, followed by gemcitabine in advanced non-small cell lung cancer: an Alpe-Adria Thoracic Oncology Multidisciplinary group study (ATOM 001). AB - Aim of this study was to determine the activity and toxicity of a sequential chemotherapy regimen in advanced non-small cell lung cancer (NSCLC). Fifty-one previously untreated stage IIIB/IV NSCLC patients were enrolled to receive two cycles of cisplatin plus paclitaxel (80/175 mg/m(2) every 21 days), followed by two cycles of vinorelbine (30 mg/m(2) on days 1 and 8 every 21 days), followed by two cycles of gemcitabine (1000 mg/m(2) on days 1, 8, and 15 every 28 days). Forty-one patients (82%) completed the planned six cycles. Grade 3-4 neutropenia was the major toxicity (41% of patients) and it was mainly associated with vinorelbine administration. Response rate after cisplatin plus paclitaxel was 18%; this percentage increased to 41% after vinorelbine, and it reached 43% upon completion of the entire six cycle treatment program. Median survival time was 14.4 months, 1-year survival rate was 53%, and 2-year survival rate was 18%. Median time to disease progression was 6.8 months. This sequential chemotherapy regimen is feasible and active in patients with advanced NSCLC. This pilot experience provides the basis for an ongoing randomized phase III trial comparing our sequential regimen versus cisplatin plus gemcitabine. PMID- 15364138 TI - A randomised phase III study of palliative radiation with concomitant carboplatin for brain metastases from non-small cell carcinoma of the lung. AB - PURPOSE: To determine if the addition of carboplatin chemotherapy to whole brain irradiation improves response and survival in patients with brain metastases from non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Forty-two patients with brain metastases from NSCLC and performance status ECOG 0-2 were randomised to receive either whole brain radiotherapy (WBRT) alone (20Gy in five fractions) or the same radiotherapy plus concomitant carboplatin (70 mg/m(2) intravenously for 5 days). RESULTS: The median survival was 4.4 months in the radiotherapy alone (RT) arm and 3.7 months in the combined treatment (RTC) arm (P = 0.64). The objective response rates of 10% on the RT arm and 29% on the RTC arm were not significantly different (P = 0.24). The trial was closed early because of poor accrual. CONCLUSIONS: Although no firm conclusions can be made regarding the efficacy of the combined treatment, this prospective study highlights the poor objective response rates and relatively poor symptom control despite standard treatment of brain metastases from NSCLC. PMID- 15364139 TI - Docetaxel and Gemcitabine administered on days 1 and 8 for metastatic non-small cell lung carcinoma (NSCLC): a phase II multicenter trial. AB - Docetaxel and Gemcitabine are active agents in non-small cell lung carcinoma (NSCLC). They have different mechanism of action, minimal overlapping toxicity, and are easily administered on an outpatient basis. This phase II study evaluated Docetaxel administered with Gemcitabine on days 1 and 8 in a 3-week cycle, to determine its efficacy, while attempting to lower the regimen's toxicity, especially myelosuppression which can occur when Docetaxel is administered at full dose on day 1 only. Forty-three chemonaive patients, 40 evaluable, were entered in this trial between May 2001 and March 2002. Thirty-seven patients had stage IV and three patients had stage III B NSCLC, median age 58 (ages 32-78), median performance status (PS) 1 (range 0-2). They were treated with Docetaxel 36mg/m(2) and Gemcitabine 1000mg/m(2) intravenously on days 1 and 8 in a 3-week cycle. No growth factors were administered. Of 40 evaluable patients, 4 achieved partial response (10%), 25 stable disease (62.5%) and 11 progressive disease (27.5%). Median time-to-disease progression was 15 weeks. Median survival was 7.75 months. One year survival was 32.5% (13 patients). Hematologic toxicity was minimal, non-hematologic toxicity was easily treatable. Docetaxel, when given with Gemcitabine on days 1 and 8 every 3 weeks, is less myelotoxic, yet still an effective treatment for metastatic NSCLC. PMID- 15364140 TI - A new induction schedule of epoetin alfa 40.000 IU in anemic patients with advanced lung cancer. AB - BACKGROUND: Non-small cell lung cancer (NSCLC) treatment with new drugs in combination with platinum salts induce anemia G1/2 and G3/4 WHO in about 35 and 10-20% of patients, respectively, with a chemotherapy (CT) dose intensity decrease in 20% of cases. Epoetin alfa, administered at standard dosages has been shown to significantly increase hemoglobin (Hb) levels, decrease transfusion requirements, and improve quality-of-life parameters in patients undergoing chemotherapy. OBJECTIVE: This open-label, non-randomized study was conducted to evaluate the efficacy and safety of an induction dose of epoetin alfa 40.000 IU in lung cancer patients with moderate or severe anemia who were receiving CT. PATIENTS AND METHODS: Twenty-four patients (8 SCLC and 16 NSCLC) were enrolled in the study to receive single subcutaneous (s.c.) injections of epoetin alfa 40.000 IU on days 1, 4, 7, 10, and 13, followed by standard treatment (10.000 IU t.i.w.) for the further 2 weeks. Nine patients had been previously treated with epoetin alfa 10.000 IU t.i.w. Twenty-two patients were receiving first-line CT and two patients were receiving docetaxel as second-line CT. RESULTS: After 15 days of treatment, in 21 evaluable patients, Hb was 10.5 +/- 1.3 g/dL (mean +/- S.D.), with a mean increase from baseline of 2.0 g/dL (95%CI: 1.3-2.7). Hb increase was > or =2g/dL in 11 patients, 1-1.9 g/dL in 5 patients, and <1g/dL in 5 patients. After 30 days of treatment, Hb was 11.5 +/- 0.8 g/dL (mean +/- S.D.), with a mean increase from baseline of 2.9 g/dL (95%CI: 2.4-3.4) in 20 evaluable patients. No adverse events possibly related to epoetin alfa treatment were observed. CONCLUSION: An induction therapy with epoetin alfa 40.000 IU for 2 weeks followed by standard treatment allows an Hb increase of 2.9 g/dL even in advanced lung cancer patients with a moderate/severe anemia, without RBC transfusion requirements. A randomized study of the proposed induction dose of epoetin alfa 40.000 IU is actually ongoing. PMID- 15364141 TI - The prognostic value of residual mediastinal involvement following induction therapy and surgery for locally advanced non-small cell lung cancer. PMID- 15364143 TI - Dexamethasone and gastrin-releasing peptide receptors in human lung cells. PMID- 15364144 TI - Can the value choices in DALYs influence global priority-setting? AB - The "disability adjusted life years" (DALYs) are increasingly used as a tool for decision-making and for describing the distribution of the Global Burden of Disease. The "DALY" combines information about mortality and morbidity, with several value choices such as disability weighting, age-weighting and discounting. These value choices imply that life years are assigned different value, depending on the age and the health state they are in. How robust is the distribution of DALYs to changes in these value choices, and are the choices transparent at the point of use? We calculated the burden attributed to "developmental disability due to malnutrition" and "major depression" with alternative value choices in a simple sensitivity analysis. In particular, we explored the relation between disability weight, health state description and incidence rate. The formulae and information needed was found in the World Health Organisation (WHO) publications using DALYs, and in a survey among international health workers. We found that alternative age-weights, disability weights and discount rate led to an inversion in the ranking of the burden of the two conditions. The DALY loss attributed to "developmental disability due to malnutrition" increased from 14 to 90%, while that of "major depression" sunk from 86 to 10%. The value choices currently used, tend at underestimating the disease burden attributed to young populations and to communicable diseases and this goes against the renewed efforts of the WHO of targeting diseases that are typical of poor populations. While the value choices may be changed, lack of transparency is a more profound problem. At the point of use, the number of DALYs attributed to a condition cannot be fully disaggregated. Hence, one cannot know which part of a DALY loss reflects the age group affected, the prevalence, the mortality rates, the disability weight assigned to it, or to how the condition has been defined. A more transparent and useful approach, we believe, would be to present the years lost due to a disease, and the years lived with a disease separately, without disability weights, age-weights and discounting. This would keep the best of the DALY approach and come closer to the aim of disentangling science from advocacy. PMID- 15364145 TI - Female genital mutilation, asylum seekers and refugees: the need for an integrated European Union agenda. AB - Asylum seekers and refugees (ASRs) are a heterogeneous population with distinct physical and psychological needs. ASRs with additional health needs are girls and women who have undergone, or are at risk of undergoing, female genital mutilation (FGM). Across the European Union (EU), variation exists in Member States' anti FGM and asylum legislation, the rigour of existing research programmes, and the operational coherence of the multiple agencies combating the practice. ASRs' needs are, consequently, not being addressed satisfactorily. This paper proposes an integrated future agenda, applicable in all EU countries, capable of meeting these girls' and women's needs. PMID- 15364146 TI - Ability to pay for maternal health services: what will it take to meet who standards? AB - High maternal morbidity and mortality in many developing countries are highly associated with poor access to and quality of health care. Here we review the economic feasibility of the WHO's mother-baby package as a means of reducing maternal and neonatal mortality and morbidity in Tanzania. This paper examines the costs of maternal health care in Tanzania, and how much can we expect households to contribute to these expenses, if the MBP were implemented. Using data from the Tanzanian 1993 Living Standard Measurement Survey (LSMS), we analyze responses from 757 women of reproductive age who have had a birth in the 12 months preceding the survey. We estimate current spending on maternal health care by different socio-economic groups and its share in relation to total household expenditures. Using logistic regression analyses, we examine the effect of the prices paid for maternal health care on the likelihood of using antenatal and safe delivery services, controlling for relevant socio-economic and demographic factors. Results show that if the MBP recovered 100% of its costs, most of the households would have to allocate more than half of their annual consumption on maternal health care. Poor socio-economic groups would experience the greatest increase in service utilization if MBP care were subsidized. In the face of scarce resources, subsidies should be targeted according to socio economic group, in order to attain equitable and sustainable maternal health services. PMID- 15364147 TI - Pricing health services for purchasers--a review of methods and experiences. AB - This article reviews methodologies and international experience related to costing and pricing health services for health care purchasers. The main factors affecting price-setting methods are: (1) provider payment systems; (2) information available on actual costs, service volumes and outcomes; and (3) characteristics of providers and purchasers. These factors are strongly interrelated. Provider payment systems determine the unit of services to be priced. In order to minimize incentives for under- or over-utilization, the prices that purchasers pay for health care services should be related to the actual unit costs of services, but accurately calculating real unit costs is intensive in terms of resources and information. Pertinent provider characteristics influencing price-setting include provider autonomy, provider negotiating power, and the degree of competition. The article presents a series of examples that run through each of these three sets of factors. The examples are from Denmark, the UK, and Thailand (for capitation); Australia, Hungary, and the United States (for case-based payment); and Germany, Korea, and Taiwan (for fee-for-service payment mechanisms). From these experiences, the article concludes with appropriate lessons for low- and middle-income countries, where the principal constraint on the development of provider payments systems is the limited availability of information on costs, volumes, and patient characteristics. PMID- 15364148 TI - Application of a single sampling plan for auditing medical-claim payments made by Taiwan National Health Insurance. AB - National Health Insurance (NHI) has been implemented in Taiwan for nearly a decade. Owing to growing demand for medical-care, effective sampling auditing is a key factor in obtaining reasonably priced medical-care services. This investigation proposes a conceptual framework "medical-claim payment auditing (MCPA) procedure" based on the "Military Standard 105E (MIL-STD-105E) single sampling plan" for establishing an objective criterion for making medical-claim payments fairly. The MCPA procedure contributes to the following: (1) meeting international standards of sampling technology; (2) reducing the sampling ratio and consequently auditing costs; and (3) encouraging healthcare providers to honestly apply their medical-claim payments thus avoid wasting medical resources. PMID- 15364149 TI - Utilization of maternal care in rural HeBei Province, the People's Republic of China: individual and structural characteristics. AB - PURPOSE: To study the effect of individual's socio-economic characteristics and the structure of the health services in the village on utilization of maternal care in rural HeBei, the People's Republic of China (PRC). DATA: Data were collected from 4273 women who gave birth to one child at least, living in a stratified sample of 288 villages in HeBei Province. FINDINGS: 54.8% of the women had at least one pre-natal care visit, 27.5% gave birth in a health care facility, and 18.1% had post-natal check-up. Utilization was inversely related to age and parity and positively to education. Occupation was related to use of pre- and post-natal services, but not to home birth. Per-capita income and living arrangement are not related to utilization. MCH worker in the village promote pre and post-natal care, but also home delivery. Village doctors promote pre-natal care and hospital delivery but do not promote post-natal check-up. Women tend to avoid the maternal services in the township health centers but some are ready to travel to city hospitals for delivery and post-natal care. CONCLUSIONS: Health education programs regarding the importance of all three maternal care services are clearly needed. These programs should address not only women of child bearing age but also care providers, MCH workers in particular. Township health center should reach-out and motivate women to use their accessible services. PMID- 15364150 TI - Willingness to pay for avoiding coronary restenosis and repeat revascularization: results from a contingent valuation study. AB - BACKGROUND: Despite technological improvements, percutaneous coronary intervention (PCI) remains limited by restenosis requiring further revascularization procedures during the ensuing year. New technologies aiming to reduce restenosis are expensive and may increase net healthcare costs. Economic evaluations of such therapies have been performed, but have been hindered by the need to assess the disutility of short-term health care events and repeat coronary revascularization as well as the lack of benchmark standards for intermediate health outcomes. The contingent valuation approach may offer particular advantages when evaluating treatments that improve short-term health outcomes. OBJECTIVE: To examine patients' willingness to pay (WTP) for treatments that may reduce the risk of restenosis and repeat revascularization after PCI. METHODS: We used a contingent valuation approach to evaluate WTP among participants in two large clinical trials evaluating new PCI devices. The baseline scenario described a 30% probability of repeat revascularization following the initial procedure. Patients were asked to indicate, using a close ended (referendum) question, their out of pocket WTP for an improved treatment that would reduce this risk. Three different prices (500 dollars, 1000 dollars, and 1500 dollars) and three levels of absolute risk reduction (10, 20, and 30%) were randomly varied creating nine sub-samples of patients. Patients' responses were analyzed using both parametric and non-parametric methods. RESULTS: 1642 patients completed the WTP question. The WTP medians for the 10 and 20% risk reductions were 273 dollars and 366 dollars, respectively; the median WTP for the 30% risk reduction was significantly higher at 1162 dollars (P<0.001). Higher household income (OR=1.57, P<0.001) was independently associated with a higher WTP. CONCLUSIONS: Although short-lived, avoidance of coronary restenosis may have considerable value to patients undergoing percutaneous coronary interventions. These findings may have important implications for emerging technologies such as drug-eluting stents. PMID- 15364151 TI - Willingness to pay for public health care: a comparison of two approaches. AB - When applying willingness-to-pay (WTP) in economic evaluations, there have been strong theoretical arguments for the use of ex ante insurance-based questions, which can be framed either as insurance premiums or taxation contributions. This paper suggests theoretical reasons why respondents may value a programme differently in these two different ex ante approaches, and inquires empirically into the potential existence of such differences. A split-sample interview study was undertaken in Denmark. The proportion of respondents willing to pay is higher in the community version, and the respondents use different reasons for being and not being willing to pay. PMID- 15364152 TI - The role of research in a technical assistance agency: the case of the 'German Agency for Technical Co-operation'. AB - Technical assistance agencies have a sustainable impact on the health systems of the countries they are operating in. As well as policy-makers at the national level, technical assistance agencies see themselves confronted that their interventions should be based on evidence, usually meaning the results of research. This study has the aim to analyse role of research in the implementation of technical assistance. We sent a questionnaire to all health project managers of the 'German Agency for Technical Co-operation' and performed a qualitative case study in one of the health projects. Forty-seven of 80 (58.8%) of the questionnaires were completed and sent back. The managers considered publications of International Organisations (IOs), scientific articles and local research as most important for their work. The case study showed application problems in the daily work. Research use not only depends on the relevance of the data but also on analytical skills, linguistic barriers and technical access to research by the potential users. The role of knowledge and information management has to be clearly defined in an organisation of technical assistance. The specific needs at the different levels have to be analysed so that skills and resources can be allocated adequately. PMID- 15364153 TI - Restaurant owner perceptions of the effects of a smoking ban. AB - In this paper business owner expectations regarding revenue changes due to a general smoking ban are analyzed using a survey mailed out to all restaurants, bars, cafes and nightclubs in Gothenburg, Sweden. A reasonable interpretation of the survey responses is that as many as 74% of the establishments do not expect to be severely hurt by a general smoking ban applying to the entire restaurant sector. The econometric results show that the owners are more likely to expect a decrease in revenues the larger their share of smoking customers. Moreover, establishments that are smoke-free are less likely to expect negative economic effects compared to those that currently allow smoking. Also, establishments with a non-smoking section are less likely to expect negative economic effects from a general smoking ban. No strong effect of the type of establishment (cafe, bar, or restaurant) on expected changes in revenues is detected, although establishments with late night hours are more likely to expect financial losses. PMID- 15364154 TI - The significance of the Cancun (non)outcome for the health sector. PMID- 15364155 TI - The effect of insulin-dependent diabetes mellitus on basal and distention-induced acid and pepsin secretion in rat. AB - BACKGROUND: Diabetes mellitus is one of the most common endocrine diseases and affects most body organs. It affects gastric acid secretion, but this effect has not been fully understood. As the effects of diabetes on gastric pepsin secretion has not been proved yet, in this experimental study basal and distension stimulated acid and pepsin secretions of diabetic and non-diabetic rats have been compared. MATERIAL AND METHODS: Female N-Mari rats weighing 200-250 g were used. Diabetic state was induced by intraperitoneal injection of 75 mg/kg streptozotocin. Animals were anaesthetized by the interaperitoneal injection of 60 mg/kg thiopental sodium. Then tracheostomy and laparotomy were done and gastric secretions were collected by a cannula entered via duodenum. Gastric distention induce by 1.5 ml normal saline per each 100g of body weight in stomach. Acid and pepsin were measured by titration and Anson's method, respectively. RESULTS: Basal gastric secretions were similar in diabetic and non diabetic animals. Distention-stimulated acid secretions in diabetic and non diabetic rats were 3.24 +/- 0.16 and 8.05 +/- 0.21 micromol/15 min, respectively, which were significantly different (P = 0.00001). Distention-induced pepsin secretion in diabetic and non diabetic rats were 3.16 +/- 0.13 and 5.24 +/- 0.16 microg/15min, respectively, which were significantly different (P = 0.00001). CONCLUSION: In this study the stomach of diabetic animals showed less reaction to distention, which may be due to the reduction of acid and pepsin secretary cells, reduction of the function of the cells, gastric atrophy or gastric vagus neuropathy. These probabilities need to be examined. PMID- 15364156 TI - Rapid adaptive down regulation of oxidative burst induced by high dextrose in human umbilical vein endothelial cells. AB - To determine whether hyperglycemia-induced increase in oxidative burst undergoes adaptive changes, the time course of superoxide (SO) production in human umbilical vein endothelial cells treated with 13.75 mM (D250) or 27.5mM dextrose (D500) was measured using the hydroethidine (HE) fluorescence method. The rate of SO production (mean +/- S.D., in arbitrary units) in cells treated with D500 during the first hour (0.758 +/- 0.367) or with D250 (0.618 +/- 0.126) was significantly higher than the rate observed in control cells treated with 100mg/dl dextrose, (D100; 0.474 +/- 0.125) (P < 0.001). However, the rate of SO production during the second, third, fourth, and fifth hour was not significantly different from that measured in control cells. The fluorescence at baseline for control cells was 3.4 +/- 2.3 and for cells treated with D500 for 1, 2, 3, 4, and 5h was 3.4 +/- 1.9, 15.2 +/- 2.5, 21.6 +/- 2.3, 27.4 +/- 3.4, and 31.8 +/- 4.3 respectively (P < 0.001). The increased baseline fluorescence suggests that the antioxidant pool may be depleted within the first hour of exposure to high concentrations of dextrose. The latter possibility is supported by the observation that treatment of cells with varying concentrations of ascorbate (15, 150, and 1500 microM) or alpha-tocopherol (10,100 and 1000 microM) prevents D500 induced increase in SO production. It is concluded that increased oxidative load in sustained chronic hyperglycemia is probably the result of depletion of antioxidant pool rather than secondary to sustained increase in SO production. PMID- 15364157 TI - Insulin secretory characteristics of monkey pancreatic islets: a simple method of islet isolation and the effect of various density gradients on separation. AB - We describe a simple stationary digestion method of islet isolation and separation by various density gradients from monkey pancreas (Macaca radiata radiata). Effective method, different types and concentrations of collagenase were standardized. Sigma type XI collagenase yielded >1000 islets/gram pancreas at the concentration of 4 mg/ml and 3 ml Hank's/gram pancreas. Slow digestion with less concentration of collagenase was suitable for monkey islet isolation. Discontinuous density gradients of bovine serum albumin (BSA) and dextran were compared with standard Ficoll for separation of islets. Islet yield (1038 +/- 81), insulin secretory response (stimulation index, S.I.11) and histological examination revealed dextran gradients were more appropriate for monkey islets when compared to BSA and Ficoll. Insulin secretory characteristics of monkey islets were studied by exposing them to low and high concentrations glucose (S.I.11.5), arginine (S.I.4.2), leucine (S.I.2.3) and tolbutamide (S.I.1.7). The results indicated that the magnitude of glucose induced insulin secretion of monkey islet is about half as that of rat and mouse islets. However, it is higher than that of porcine and bovine islets. In conclusion, the knowledge of insulin secretory ability of Indian bonnet monkey islets together with the techniques of isolation and separation are useful tool for diabetic research especially islet transplantation. PMID- 15364158 TI - Postprandial versus preprandial dosing of biphasic insulin aspart in elderly type 2 diabetes patients. AB - Preprandial dosing (within 5 min before meal) and postprandial dosing (15-20 min after meal onset) of NovoLog Mix 70/30 (BIAsp 30, a biphasic formulation of insulin aspart, 30% soluble and 70% protamine-crystallized) were compared in elderly (> or =65 years) type 2 diabetes patients in this open-label, 12-week, crossover study. Ninety-three patients were treated with b.i.d. preprandial injections of BIAsp 30 during a 2-week run-in period and subsequently randomized to a 4-week treatment with either pre- or postprandial b.i.d. BIAsp 30, followed by crossover to the other regimen for 4 weeks. Mean plasma glucose values during a 4-h mealtest at the end of each treatment were similar for pre- and postprandial BIAsp 30 (153 +/- 58 mg/dl and 161 +/- 59 mg/dl, respectively, difference not significant). However, the mean blood glucose increment from self measured blood glucose values was slightly but significantly greater after postprandial injection than after preprandial injection (treatment difference: 16.3mg/dl; 95% CI: [0.5, 29.3]). Fifty-six percent of patients reported a hypoglycemic episode; postprandial injection did not increase the incidence of hypoglycemia as compared to preprandial injection (113 episodes versus 125 episodes, respectively). For some elderly type 2 diabetes patients, postprandial injection of BIAsp 30 may be an acceptable alternative to standard preprandial injection. PMID- 15364159 TI - Acute-phase proteins and microalbuminuria among patients with type 2 diabetes. AB - OBJECTIVE: to determine the relationship between acute-phase proteins and microalbuminuria in type 2 diabetic patients without clinical evidence of macrovascular disease. RESEARCH DESIGN AND METHODS: We studied cross-sectionally 64 non-smoking outpatients with type 2 diabetes mellitus without clinical evidence of cardiovascular disease attended at Brazilian University General Hospital aged 59.5 +/- 8.1 years and with a known duration of diabetes of 11.5 +/ 8 years. Urinary albumin excretion rate (AER) was determined in timed overnight urine samples. Serum alpha(1)-acid glycoprotein (AGP) and plasma fibrinogen were determined by immunoturbidimetry assay and serum C-reactive protein (CRP) was measured by a high-sensitive immunonephelometry assay. RESULTS: A higher levels of AGP (P = 0.0000), CRP (P= 0.003) and fibrinogen ( P = 0.0001) were found in microalbuminuric (n = 26) than in normoalbuminuric patients ( n = 38). All the acute-phase proteins were correlated with each other and with AER, respectively (r = 0.67 for AGP; 0.35 for fibrinogen, and 0.41 for CRP, P < 0.01 for all). Stepwise multiple regression analysis showed that AGP was independently associated with AER along with systolic blood pressure (r2 = 0.49, P = 0.000). Logistic regression analysis showed that AGP was independently related to microalbuminuria with an odds ratio (95% CI) of 1.16 ((1.08-1.24), P = 0.000). CONCLUSIONS: According to our results acute-phase proteins a known markers of chronic inflammation were associated with microalbuminuria independently of clinical cardiovascular disease. The influence of AGP on AER and microalbuminuria needs to be confirmed in prospective studies. Intervention studies are necessary to assess whether anti-inflammatory treatment would have a beneficial effect on this chronic complication of diabetes. PMID- 15364160 TI - Association of serum apolipoprotein C III levels and apolipoprotein C III gene Sst I polymorphism with carotid intima-media thickness in Chinese type 2 diabetic patients. AB - Apolipoprotein C III (apo C III) plays a central role in regulating plasma metabolism of triglyceride-rich lipoprotein (TRL). The G3238C allele (Sst I) in the 3'-untranslated region has been found to be associated with raised apo C III levels and hypertriglyceridemia (HTG). Some studies suggest that apo C III and the S(2) allele of apo C III gene are independent risk factors for atherosclerotic diseases. To study the potential association between these factors we analyzed the clinical data and their correlations with serum apo C III levels, apo C III gene Sst I polymorphism, and carotid intima-media thickness (IMT) in 78 unrelated Chinese patients with type 2 diabetes. Apo C III gene Sst I polymorphism was examined using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Carotid IMT was measured by color doppler ultrasound examination. Serum apo C III levels were found to be positively associated with plasma TG ( r = 0.527, P < 0.001), TC (r = 0.424, P < 0.001), LDL C (r = 0.308, P < 0.01) concentrations, and carotid IMT (r =0.359, P < 0.01 ). Multivariate analysis (backward) showed that diastolic blood pressure, apo C III, and fasting insulin levels were independent risk factors of carotid IMT. However, the results did not show the association between S(2) allele and carotid IMT in our diabetic patients. Thus, our study suggested that apo C III is an independent risk factor for atherosclerotic diseases in Chinese type 2 diabetes. PMID- 15364161 TI - Improved glycemic control without an increase in severe hypoglycemic episodes in intensively treated patients with type 1 diabetes receiving morning, evening, or split dose insulin glargine. AB - OBJECTIVE: To see if insulin glargine improves glycemic control in a clinical setting. RESEARCH DESIGN AND METHODS: A questionnaire and electronic database were used to assess glycemic parameters for 292 type 1 diabetic subjects taking > or =4 injections per day and receiving glargine as their only long-acting basal insulin for at least 6 months. Sixty-three subjects were taking glargine in the morning, 125 were taking glargine in the evening, and 104 were splitting the glargine dose between the morning and evening. RESULTS: The mean (+/-S.D.) age and duration of diabetes were 32 +/- 10 years and 15.9 +/- 10.3 years, respectively. The mean (+/-S.E.M.) durations of treatment with glargine were 13.1 +/- 0.6 months, 12.2 +.- 0.4 months, and 14.3 +/- 0.5 months for the morning, evening, and split treatment groups, respectively (P < 0.01). The A1C values improved significantly from baseline for the evening and the split dosage groups or when all groups were combined. The mean basal insulin dose was significantly reduced at the end of the study in all the three groups from baseline with no change in the short-acting insulin dose. The number of severe hypoglycemic episodes decreased from 379 in the year prior to glargine treatment to 167 in the post-glargine year. The weight gain was significantly higher in the group that took the split glargine dose (P < 0.01). CONCLUSIONS: Similar or improved glycemic control was achieved by administering glargine in the morning, evening, or using a split dose without any further increase in severe hypoglycemic episodes. Splitting the glargine dose did not offer any advantages in glycemic control parameters. PMID- 15364162 TI - Relationship between brachial-ankle pulse wave velocity and cardiovascular risk factors of the metabolic syndrome. AB - In this study, we investigated the association between brachial-ankle pulse wave velocity (baPWV), a novel non-invasive means of measuring atherosclerosis, and the cardiovascular risk factors of the metabolic syndrome. The study group comprised of 368 Koreans without history of diabetes or hypertension. Anthropometric parameters, blood pressure, fasting blood glucose (FBG), lipid profiles, ankle-brachial pressure index (ABI), and baPWV were measured in each subject. ABI and baPWV levels were higher in men than in women. ABI and baPWV levels were not correlated with each other. In women, baPWV was closely associated with the cardiovascular risk factors of the metabolic syndrome. Women with the metabolic syndrome showed higher baPWV levels compared to those without the metabolic syndrome (1540 +/- 281 versus 1312 +/- 223, P < 0.001). Multiple regression analysis showed that baPWV was significantly associated with systolic blood pressure, age, sex, waist circumference, and FBG (adjusted R2 = 0.548). The present study shows that baPWV is significantly associated with the features of the metabolic syndrome in Korean women. PMID- 15364163 TI - Five missense mutations in glucagon-like peptide 1 receptor gene in Japanese population. AB - To address the possibility that the partial disruption of Glucagon-like peptide-1 (GLP-1) signaling could cause diabetes, we tried to detect the mutation in GLP-1 receptor (GLP-1R) gene in the population with type 2 diabetes. Genomic DNA was extracted from 36 unrelated Japanese type 2 diabetic subjects and directly sequenced for the GLP-1R gene. For the detected polymorphisms, 791 patients with type 2 diabetes and 318 controls were screened by polymerase chain reaction restricted fragment length polymorphism and association study was carried out. Five missense and four silent variants were detected in the GLP-1R gene. There were no significant differences in the frequencies of Pro7Leu, Arg44His and Leu260Pro polymorphism between the diabetic and control groups. And also there were no significant differences in body mass index (BMI), onset age and fasting IRI among the wild type, heterozygote and homozygote of these variants in diabetic patients. Thr149Met mutation was detected in one case among 791 type 2 diabetes patients, but not in control subjects. The patient with this mutation exhibited impairment of both insulin secretion, insulin sensitivity and glucose effectiveness, which may be partially explained by Thr149Met mutation in GLP-1R, though family linkage analysis and function analysis remain to be examined. PMID- 15364164 TI - The relationship of health beliefs and complication prevention behaviors of Chinese individuals with Type 2 Diabetes Mellitus. AB - This study aimed to identify the relationship of health beliefs and complication prevention behaviors among Chinese individuals with Type 2 Diabetes Mellitus in Malaysia. A correlation study using the Health Belief Model (HBM) framework was undertaken with 128 Chinese subjects with Type 2 Diabetes of both gender, mean age 60.5 +/- 8.42 years from one urban hospital and four rural health centers. Research tool was a 60-item questionnaire with responses recorded on 5-point Likert scale. Data was analyzed with descriptive statistics, Spearmen correlation and Mann-Whitney U-test. The majority of the subjects had less than 6 years of education. Seventy-two percent of them were aware of diabetes complications and its risk factors. However, few subjects practiced complication preventive measures because of lack of perceived seriousness of diabetes and lack of perceived susceptibility to diabetes complications. There were significant correlations between complication prevention behaviors and perceived severity (P < 0.05), perceived susceptibility (P < 0.05 ) and perceived barrier (P < 0.05 ); subjects' education level and perceived severity (P < 0.05), perceived susceptibility (P < 0.05) and complication prevention behavior (P < 0.05). There was no significant correlation between health beliefs and settings; genders; disease duration and treatment mode. In conclusion, poor complication preventive behavior among the subjects was associated with lack of perceived seriousness of diabetes and lack of perceived susceptibility to diabetes complications. PMID- 15364165 TI - Confirmation of a susceptibility locus for diabetic nephropathy on chromosome 3q23-q24 by association study in Russian type 1 diabetic patients. AB - Family-based studies and segregation analyses suggest that inherited factors play a significant role in susceptibility to diabetic nephropathy (DN). Moczulski et al. [Diabetes 47 (1998) 1164-1169] found a susceptibility locus for DN in type 1 diabetes covering a 20cM region on chromosome 3q, with a peak of linkage close to the angiotensin II type 1 receptor (AT1) gene. We examined eight polymorphic markers (D3S1512, D3S1550, D3S1557, D3S1744, D3S2326, D3S3599, D3S3694, and a (CA)(n) dinucleotide repeat polymorphism in the 3' flanking region of the AT1 gene) spanning about 6.2 megabases (Mb) in the region of maximal linkage with DN on chromosome 3q23-q24. The markers were used to genotype a total of 381 Russian type 1 diabetic subjects, 195 of whom had DN and 186 had no clinical nephropathy. Four of the markers tested, D3S1512, D3S1550, D3S2326, and D3S3599, showed an association with DN in type 1 diabetes mellitus. These markers are located within a 1.0Mb interval that starts about 4.4Mb centromeric to the AT1 gene. Thus, our results suggest the existence of the DN susceptibility locus previously described by Moczulski et al. on chromosome 3q. PMID- 15364166 TI - Antiproteinuric effect of candesartan cilexetil in Japanese subjects with type 2 diabetes and nephropathy. AB - The effect of the angiotensin II receptor blocker, candesartan cilexetil, on proteinuria was examined in a prospective, multicenter, randomized, double-blind study in Japanese subjects with type 2 diabetes. This study enrolled diabetic subjects with confirmed proteinuria into four groups for 12 weeks of treatment with placebo or candesartan cilexetil 2, 4, or 8 mg. The contribution of the angiotensin converting enzyme (ACE) gene polymorphism to the effect of candesartan cilexetil was also examined. In 127 subjects, candesartan cilexetil showed a dose-related reduction in proteinuria after 12 weeks of treatment (F = 9.45, P = 0.0013), with a 18.1% reduction in the 4-mg group, and a 5.8% reduction in the 8-mg group, in contrast to a 32.2% increase in the placebo group, and a 0.8% increase in the 2-mg group. These results indicate that candesartan cilexetil is useful in reducing proteinuria in diabetic subjects when compared with placebo. In addition, candesartan cilexetil seems to be effective in subjects with both the II and DD genotypes of the ACE gene. PMID- 15364167 TI - A comparative study of atorvastatin and simvastatin as monotherapy for mixed hyperlipidaemia in Type 2 diabetic patients. PMID- 15364168 TI - China must prioritise health opportunities for all. PMID- 15364169 TI - Choose advice wisely for health. PMID- 15364170 TI - Clinical trial registration: a statement from the International Committee of Medical Journal Editors. PMID- 15364171 TI - How can cross-country research on health risks strengthen interventions? Lessons from INTERHEART. PMID- 15364172 TI - Influencing birth outcomes in Nepal. PMID- 15364173 TI - Can radiation therapy for breast cancer be avoided? PMID- 15364174 TI - Can the genotoxicity of chemotherapy be predicted? PMID- 15364175 TI - The Global Fund's principal recipients... or neglected partners. PMID- 15364176 TI - Standards of care in the antiretroviral rollout world. PMID- 15364177 TI - VALUE: analysis of results. PMID- 15364178 TI - VALUE: analysis of results. PMID- 15364179 TI - VALUE: analysis of results. PMID- 15364180 TI - VALUE: analysis of results. PMID- 15364181 TI - VALUE: analysis of results. PMID- 15364182 TI - VALUE: analysis of results. PMID- 15364184 TI - History of thymoma and yellow fever vaccination. PMID- 15364185 TI - Effect of potentially modifiable risk factors associated with myocardial infarction in 52 countries (the INTERHEART study): case-control study. AB - BACKGROUND: Although more than 80% of the global burden of cardiovascular disease occurs in low-income and middle-income countries, knowledge of the importance of risk factors is largely derived from developed countries. Therefore, the effect of such factors on risk of coronary heart disease in most regions of the world is unknown. METHODS: We established a standardised case-control study of acute myocardial infarction in 52 countries, representing every inhabited continent. 15152 cases and 14820 controls were enrolled. The relation of smoking, history of hypertension or diabetes, waist/hip ratio, dietary patterns, physical activity, consumption of alcohol, blood apolipoproteins (Apo), and psychosocial factors to myocardial infarction are reported here. Odds ratios and their 99% CIs for the association of risk factors to myocardial infarction and their population attributable risks (PAR) were calculated. FINDINGS: Smoking (odds ratio 2.87 for current vs never, PAR 35.7% for current and former vs never), raised ApoB/ApoA1 ratio (3.25 for top vs lowest quintile, PAR 49.2% for top four quintiles vs lowest quintile), history of hypertension (1.91, PAR 17.9%), diabetes (2.37, PAR 9.9%), abdominal obesity (1.12 for top vs lowest tertile and 1.62 for middle vs lowest tertile, PAR 20.1% for top two tertiles vs lowest tertile), psychosocial factors (2.67, PAR 32.5%), daily consumption of fruits and vegetables (0.70, PAR 13.7% for lack of daily consumption), regular alcohol consumption (0.91, PAR 6.7%), and regular physical activity (0.86, PAR 12.2%), were all significantly related to acute myocardial infarction (p<0.0001 for all risk factors and p=0.03 for alcohol). These associations were noted in men and women, old and young, and in all regions of the world. Collectively, these nine risk factors accounted for 90% of the PAR in men and 94% in women. INTERPRETATION: Abnormal lipids, smoking, hypertension, diabetes, abdominal obesity, psychosocial factors, consumption of fruits, vegetables, and alcohol, and regular physical activity account for most of the risk of myocardial infarction worldwide in both sexes and at all ages in all regions. This finding suggests that approaches to prevention can be based on similar principles worldwide and have the potential to prevent most premature cases of myocardial infarction. PMID- 15364186 TI - Association of psychosocial risk factors with risk of acute myocardial infarction in 11119 cases and 13648 controls from 52 countries (the INTERHEART study): case control study. AB - BACKGROUND: Psychosocial factors have been reported to be independently associated with coronary heart disease. However, previous studies have been in mainly North American or European populations. The aim of the present analysis was to investigate the relation of psychosocial factors to risk of myocardial infarction in 24767 people from 52 countries. METHODS: We used a case-control design with 11119 patients with a first myocardial infarction and 13648 age matched (up to 5 years older or younger) and sex-matched controls from 262 centres in Asia, Europe, the Middle East, Africa, Australia, and North and South America. Data for demographic factors, education, income, and cardiovascular risk factors were obtained by standardised approaches. Psychosocial stress was assessed by four simple questions about stress at work and at home, financial stress, and major life events in the past year. Additional questions assessed locus of control and presence of depression. FINDINGS: People with myocardial infarction (cases) reported higher prevalence of all four stress factors (p<0.0001). Of those cases still working, 23.0% (n=1249) experienced several periods of work stress compared with 17.9% (1324) of controls, and 10.0% (540) experienced permanent work stress during the previous year versus 5.0% (372) of controls. Odds ratios were 1.38 (99% CI 1.19-1.61) for several periods of work stress and 2.14 (1.73-2.64) for permanent stress at work, adjusted for age, sex, geographic region, and smoking. 11.6% (1288) of cases had several periods of stress at home compared with 8.6% (1179) of controls (odds ratio 1.52 [99% CI 1.34-1.72]), and 3.5% (384) of cases reported permanent stress at home versus 1.9% (253) of controls (2.12 [1.68-2.65]). General stress (work, home, or both) was associated with an odds ratio of 1.45 (99% CI 1.30-1.61) for several periods and 2.17 (1.84-2.55) for permanent stress. Severe financial stress was more typical in cases than controls (14.6% [1622] vs 12.2% [1659]; odds ratio 1.33 [99% CI 1.19-1.48]). Stressful life events in the past year were also more frequent in cases than controls (16.1% [1790] vs 13.0% [1771]; 1.48 [1.33-1.64]), as was depression (24.0% [2673] vs 17.6% [2404]; odds ratio 1.55 [1.42-1.69]). These differences were consistent across regions, in different ethnic groups, and in men and women. INTERPRETATION: Presence of psychosocial stressors is associated with increased risk of acute myocardial infarction, suggesting that approaches aimed at modifying these factors should be developed. PMID- 15364187 TI - MMR vaccination and pervasive developmental disorders: a case-control study. AB - BACKGROUND: Concern that measles-mumps-rubella (MMR) vaccination might cause autism has led to a fall in vaccine coverage. We investigated whether MMR vaccination is associated with an increased risk of autism or other pervasive developmental disorders. METHODS: We did a matched case-control study using the UK General Practice Research Database. Cases were people born in 1973 or later who had first recorded diagnosis of pervasive developmental disorder while registered with a contributing general practice between 1987 and 2001. Controls were matched on age, sex, and general practice. FINDINGS: 1294 cases and 4469 controls were included. 1010 cases (78.1%) had MMR vaccination recorded before diagnosis, compared with 3671 controls (82.1%) before the age at which their matched case was diagnosed. After adjustment for age at joining the database, the odds ratio for association between MMR and pervasive developmental disorder was 0.86 (95% CI 0.68-1.09). Findings were similar when restricted to children with a diagnosis of autism, to those vaccinated with MMR before the third birthday, or to the period before media coverage of the hypothesis linking MMR with autism. INTERPRETATION: Our findings suggest that MMR vaccination is not associated with an increased risk of pervasive developmental disorders. PMID- 15364188 TI - Effect of a participatory intervention with women's groups on birth outcomes in Nepal: cluster-randomised controlled trial. AB - BACKGROUND: Neonatal deaths in developing countries make the largest contribution to global mortality in children younger than 5 years. 90% of deliveries in the poorest quintile of households happen at home. We postulated that a community based participatory intervention could significantly reduce neonatal mortality rates. METHODS: We pair-matched 42 geopolitical clusters in Makwanpur district, Nepal, selected 12 pairs randomly, and randomly assigned one of each pair to intervention or control. In each intervention cluster (average population 7000), a female facilitator convened nine women's group meetings every month. The facilitator supported groups through an action-learning cycle in which they identified local perinatal problems and formulated strategies to address them. We monitored birth outcomes in a cohort of 28?931 women, of whom 8% joined the groups. The primary outcome was neonatal mortality rate. Other outcomes included stillbirths and maternal deaths, uptake of antenatal and delivery services, home care practices, infant morbidity, and health-care seeking. Analysis was by intention to treat. The study is registered as an International Standard Randomised Controlled Trial, number ISRCTN31137309. FINDINGS: From 2001 to 2003, the neonatal mortality rate was 26.2 per 1000 (76 deaths per 2899 livebirths) in intervention clusters compared with 36.9 per 1000 (119 deaths per 3226 livebirths) in controls (adjusted odds ratio 0.70 [95% CI 0.53-0.94]). Stillbirth rates were similar in both groups. The maternal mortality ratio was 69 per 100000 (two deaths per 2899 livebirths) in intervention clusters compared with 341 per 100000 (11 deaths per 3226 livebirths) in control clusters (0.22 [0.05-0.90]). Women in intervention clusters were more likely to have antenatal care, institutional delivery, trained birth attendance, and hygienic care than were controls. INTERPRETATION: Birth outcomes in a poor rural population improved greatly through a low cost, potentially sustainable and scalable, participatory intervention with women's groups. PMID- 15364189 TI - Finger pricking. PMID- 15364190 TI - Antenatal anthelmintic treatment, birthweight, and infant survival in rural Nepal. AB - Anthelmintic treatment, which is recommended during pregnancy in areas where there is a high rate of anaemia, needs further investigation. We examined prospectively the association between anthelmintic treatment and maternal anaemia, birthweight, and infant mortality in a study of prenatal supplements, in which women received albendazole twice during pregnancy. Women given albendazole in the second trimester of pregnancy had a lower rate of severe anaemia during the third trimester. Birthweight of infants of women who had received two doses of albendazole rose by 59 g (95% CI 19-98), and infant mortality at 6 months fell by 41% (RR 0.59; 95% CI 0.43-0.82). Antenatal anthelmintics could be effective in reducing maternal anaemia and improving birthweight and infant survival in hookworm-endemic regions. PMID- 15364191 TI - Being wrong in the right direction? PMID- 15364192 TI - Prospects for new drugs for chronic obstructive pulmonary disease. AB - No currently available treatments have been shown to slow the progression of chronic obstructive pulmonary disease (COPD) or suppress the inflammation in small airways and lung parenchyma. However, several new treatments are in clinical development; some target the inflammatory process and others are directed against structural cells. A group of specific therapies are directed against the influx of inflammatory cells into the airways and lung parenchyma that occurs in COPD; these include agents directed against adhesion molecules and chemokines, as well as therapies to oppose tumour necrosis factor alpha and increase interleukin 10. Broad-range anti-inflammatory drugs are now in phase III development for COPD; they include inhibitors of phosphodiesterase 4. Other drugs that inhibit cell signalling include inhibitors of p38 mitogen-activated protein kinase, nuclear factor kappaB, and phosphoinositide-3-kinase gamma. More specific approaches are to give antioxidants, inhibitors of inducible nitric oxide synthase, and antagonists of leukotriene B4 receptor. Inhibitors of epidermal growth-factor-receptor kinase and calcium-activated chloride channels have the potential to prevent overproduction of mucus. Therapy to inhibit fibrosis is being developed against transforming growth factor beta1 and protease-activated receptor 2. There is also a search for inhibitors of serine proteinases and matrix metalloproteinases to prevent lung destruction and the development of emphysema, as well as drugs such as retinoids that might even reverse this process. Effective delivery of drugs to the sites of disease in the peripheral lung is an important consideration, and there is a need for validated biomarkers and monitoring techniques in early clinical studies with new therapies for COPD. PMID- 15364193 TI - Informed choices for attaining the Millennium Development Goals: towards an international cooperative agenda for health-systems research. AB - Health systems constraints are impeding the implementation of major global initiatives for health and the attainment of the Millennium Development Goals (MDGs). Research could contribute to overcoming these barriers. An independent task force has been convened by WHO to suggest areas where international collaborative research could help to generate the knowledge necessary to improve health systems. Suggested topics encompass financial and human resources, organisation and delivery of health services, governance, stewardship, knowledge management, and global influences. These topics should be viewed as tentative suggestions that form a basis for further discussion. This article is part of a wide-ranging consultation and comment is invited. The potential agenda will be presented at the Ministerial Summit on Health Research in November, 2004, and revised in the light of responses. Subsequently, we hope that resources will be committed to generate the evidence needed to build the equitable, effective, and efficient health systems needed to achieve the MDGs PMID- 15364194 TI - The state of primary-care research. PMID- 15364195 TI - Low dose synthetic prostaglandin E2: always safe for pregnancy termination? PMID- 15364196 TI - Cell and molecular biology of spindle poles and NuMA. AB - Mitotic and meiotic cells contain a bipolar spindle apparatus of microtubules and associated proteins. To arrange microtubules into focused spindle poles, different mechanisms are used by various organisms. Principally, two major pathways have been characterized: nucleation and anchorage of microtubules at preexisting centers such as centrosomes or spindle pole bodies, or microtubule growth off the surface of chromosomes, followed by sorting and focusing into spindle poles. These two mechanisms can even be found in cells of the same organism: whereas most somatic animal cells utilize the centrosome as an organizing center for spindle microtubules, female meiotic cells build an acentriolar spindle apparatus. Most interestingly, the molecular components that drive acentriolar spindle pole formation are also present in cells containing centrosomes. They include microtubule-dependent motor proteins and a variety of structural proteins that regulate microtubule orientation, anchoring, and stability. The first of these spindle pole proteins, NuMA, had already been identified more than 20 years ago. In addition, several new proteins have been characterized more recently. This review discusses their role during spindle formation and their regulation in the cell cycle. PMID- 15364197 TI - Organelle nuclei in higher plants: structure, composition, function, and evolution. AB - Plant cells have two distinct types of energy-converting organelles: plastids and mitochondria. These organelles have their own DNAs and are regarded as descendants of endosymbiotic prokaryotes. The organelle DNAs associate with various proteins to form compact DNA-protein complexes, which are referred to as organelle nuclei or nucleoids. Various functions of organelle genomes, such as DNA replication and transcription, are performed within these compact structures. Fluorescence microscopy using the DNA-specific fluorochrome 4',6-diamidino-2 phenylindole has played a pivotal role in establishing the concept of "organelle nuclei." This fluorochrome has also facilitated the isolation of morphologically intact organelle nuclei, which is indispensable for understanding their structure and composition. Moreover, development of an in vitro transcription?DNA synthesis system using isolated organelle nuclei has provided us with a means of measuring and analyzing the function of organelle nuclei. In addition to these morphological and biochemical approaches, genomics has also had a great impact on our ability to investigate the components of organelle nuclei. These analyses have revealed that organelle nuclei are not a vestige of the bacterial counterpart, but rather are a complex system established through extensive interaction between organelle and cell nuclear genomes during evolution. Extensive diversion or exchange during evolution is predicted to have occurred for several important structural proteins, such as major DNA-compacting proteins, and functional proteins, such as RNA and DNA polymerases, resulting in complex mechanisms to control the function of organelle genomes. Thus, organelle nuclei represent the most dynamic front of interaction between the three genomes (cell nuclear, plastid, and mitochondrial) constituting eukaryotic plant cells. PMID- 15364198 TI - The centrosome in higher organisms: structure, composition, and duplication. AB - The centrosome found in higher organisms is an organelle with a complex and dynamic architecture and composition. This organelle not only functions as a microtubule-organizing center, but also is integrated with or impacts a number of cellular processes. Defects associated with this organelle have been linked to a variety of human diseases including several forms of cancer. Here we review the emerging picture of how the structure, composition, duplication, and function of the centrosome found in higher organisms are interrelated. PMID- 15364199 TI - Functions of myc:max in the control of cell proliferation and tumorigenesis. AB - Deregulation and elevated expression of members of the Myc family of bHLHZip transcription factors are observed in a high percentage of tumors. This close association with human cancers has led to a tremendous effort to define their biological and biochemical activities. Although Myc family proteins have the capacity to elicit a wide range of cell behaviors, their principal function appears to be to drive cells into the cell cycle and to keep them there. However, forced expression of Myc profoundly sensitizes normal cells to apoptosis. Therefore, tumor formation caused by deregulated Myc expression requires cooperating events that disrupt pathways that mediate apoptosis. Myc-dependent tumor formation may also be impeded by a set of related bHLHZip proteins with the demonstrated potential to act as Myc antagonists in cell culture experiments. In this review, we examine the complex activities of Myc family proteins and how their actions might be regulated in the context of a network of bHLHZip proteins. PMID- 15364200 TI - Electrophysiological approaches to the study of protein translocation in mitochondria. AB - Electrophysiological techniques have been integral to our understanding of protein translocation across various membranes, and, in particular, the mitochondrial inner and outer membranes. Descriptions of various methodologies (for example, patch clamp, planar bilayers, and tip dip, and their past and potential contributions) are detailed within. The activity of protein import channels of native mitochondrial inner and outer membranes can be studied by directly patch clamping mitochondria and mitoplasts (mitochondria stripped of their outer membrane by French pressing) from various genetically manipulated strains of yeast and mammalian tissue cultured cells. The channel activities of TOM, TIM23, and TIM22 complexes are compared with those reconstituted in proteoliposomes and with those of the recombinant proteins Tom40p, Tim23p, and Tim22p, which play major roles in protein translocation. Studies of the mechanism(s) and the role of channels in protein translocation in mitochondria are prototypes, as the same principles are likely followed in all biological membranes including the endoplasmic reticulum and chloroplasts. The ability to apply electrophysiological techniques to these channels is now allowing investigations into the role of mitochondria in diverse fields such as neurotransmitter release, long-term potentiation, and apoptosis. PMID- 15364201 TI - Use of fecal glucocorticoid metabolite measures in conservation biology research: considerations for application and interpretation. AB - Fecal glucocorticoid metabolite analyses are increasingly being used by a variety of scientists (e.g., conservation biologists, animal scientists) to examine glucocorticoid (i.e., stress hormone) secretion in domestic and wild vertebrates. Adrenocortical activity (i.e., stress response) is of interest to conservation biologists because stress can alter animal behavior, reduce resistance to disease, and affect population performance. The noninvasiveness of fecal-based assessments is attractive, particularly when studying endangered species, because samples can often be obtained without disturbing the animal. Despite such advantages, many confounding factors inhibit the utility of this technique in addressing conservation problems. In particular, interpretation of fecal glucocorticoid metabolite (FGM) measures may be confounded by the length of time animals are held in captivity, normal seasonal and daily rhythms, body condition, sample storage and treatment techniques, diet of the animal, assay selection, animal status (i.e., social ranking, reproductive status), sample age and condition, and sample mass. Further complicating interpretation and utility of these measures is the apparent species-specific response to these factors. The purpose of this paper is to discuss the factors that confound interpretation of FGM measures, summarize research that addresses these issues, and offer an agenda for future research and interpretation. We urge conservation biologists to carefully consider confounding factors and the relationship between FGM secretion and population performance and biological costs when investigating effects of environmental and human-induced disturbances on wildlife. The crisis nature of many decisions in conservation biology often requires decisions from limited data; however, confirmatory results should not be posited when data are incomplete or confounding factors are not understood. Building reliable databases, and research with surrogate species when possible, will aid future efforts and enhance the utility of FGM assays. PMID- 15364202 TI - Molecular cloning of bullfrog prolactin receptor cDNA: changes in prolactin receptor mRNA level during metamorphosis. AB - In amphibian larvae, prolactin (PRL) is known to possess growth-promoting and anti-metamorphic activities. For further understanding of the role of PRL in larvae, bullfrog PRL receptor (bfPRLR) cDNA was obtained from the tail fin of premetamorphic tadpoles by use of the reverse transcription-polymerase chain reaction (RT-PCR) coupled with 5' - and 3' -rapid amplification of cDNA ends (RACE). The predicted bfPRLR was composed of 617 amino acids, contained a single transmembrane domain, and showed 33-57% sequence homologies with known sequences of vertebrate PRLRs. When bfPRLR was transiently expressed, specific binding of 125I-labeled bullfrog PRL (bfPRL) was observed. By Northern blot analysis, a 3-kb transcript was detected in the tail fin. By RT-PCR bfPRLR mRNA expression was detected mainly in the brain, kidney, skin, and tail throughout prometamorphic and middle climactic periods. The results of an RNase protection assay revealed that the bfPRLR mRNA level in the tail fin increased around the onset of climax (stage XX) and was maintained at a relatively high value at least until mid climax (stage XXII). It also revealed that bfPRLR mRNA level in the kidney of larvae gradually rose as metamorphosis progressed. The results support the view that PRL in larval period acts not only on the larval organs but also on the organs that are necessary for the adult life to maintain or develop their structures and functions. PMID- 15364203 TI - Female pheromones stimulate release of luteinizing hormone and testosterone without altering GnRH mRNA in adult male Syrian hamsters (Mesocricetus auratus). AB - In many species chemosensory stimuli function as important signals that influence reproductive status. Neurons synthesizing the peptide gonadotropin-releasing hormone (GnRH) are critical mediators of reproductive function via their regulation of the hypothalamic-pituitary-gonadal (HPG) axis, and they are thought to be responsive to chemosensory information. In the present study, we sought to elucidate the effects of female chemosensory stimuli on the HPG axis in sexually naive adult male Syrian hamsters. In Experiment 1, serial blood samples were collected from catheterized male hamsters following exposure to female pheromones in order to characterize the luteinizing hormone (LH) response to this chemosensory stimulus. In Experiment 2, brains and terminal blood samples were collected from animals 0, 60, and 120 min following pheromone exposure. GnRH mRNA was measured in brain tissue sections using in situ hybridization, and plasma concentrations of LH and testosterone were measured using radioimmunoassay. Data from Experiment 1 indicated that female pheromones elicited a rapid rise in plasma LH that peaked at 15 min and returned to baseline 45 min after exposure. In Experiment 2, testosterone was elevated in terminal blood samples obtained 60 min, but not 120 min, after exposure to pheromones. LH levels were unaffected at both of these time points. The chemosensory-induced increases in LH and testosterone release were not accompanied by subsequent changes in GnRH mRNA over the time course studied. These data suggest that while activation of the male HPG axis by female pheromones involves release of GnRH, it does not involve increases in GnRH mRNA 1-2 h after pheromonal stimulation as a mechanism for replenishment of released peptide. PMID- 15364204 TI - Molecular cloning of bullfrog corticotropin-releasing factor (CRF): effect of homologous CRF on the release of TSH from pituitary cells in vitro. AB - Corticotropin-releasing factor (CRF) plays multiple roles in vertebrate species. In non-mammalian vertebrates, CRF has been considered to be the major thyrotropin (TSH)-releasing factor. This notion, however, was derived from experimental data on CRF of mammalian origin. Moreover, in the case of amphibians it has never been directly proved that CRF stimulates the release of TSH from the pituitary. The presently described experiment was conducted to provide direct evidence that homologous CRF enhances the release of TSH from the bullfrog (Rana catesbeiana) pituitary. First, cloning of cDNA encoding bullfrog CRF (fCRF) was accomplished. The cDNA encoding fCRF precursor was isolated from a cDNA library of the bullfrog hypothalamus. The amino acid sequence of fCRF predicted from the amplified cDNA sequence showed 83 and 95% identities with the sequences of ovine and human CRFs, respectively. An antiserum against the fCRF synthesized on the basis of the amino acid sequence was raised and used for immunohistochemical staining of the hypothalamus-hypophyseal region of the bullfrog brain. It stained some of the cell bodies situated mainly in the preoptic area, the nucleus infundibularis dorsalis and nucleus hypothalamicus ventralis and the axons that terminate in the median eminence and neural lobe. The synthetic fCRF was tested for its TSH releasing activity toward anterior pituitary cells of adult bullfrogs in an in vitro system. As a result, the fCRF caused the release of TSH from the dispersed pituitary cells into the culture medium concentration-dependently, as measured by a specific radioimmunoassay for bullfrog TSH. The potency of the fCRF was almost equivalent to that of ovine CRF. Human urocortin III (hUCN III), a CRF receptor type 2 (CRF-R2) specific agonist enhanced the release of TSH from the pituitary cells in culture, suggesting the involvement of CRF-R2 in the CRF-induced TSH release in the bullfrogs. Culture of pituitary cells in the presence of the hypothalamic extract (HE) and alpha-helical CRF(9-41), a CRF-R antagonist, revealed that the antagonist suppressed the TSH-releasing activity of the HE by approximately 50%, suggesting that endogenous CRF contributes as a TSH-releasing factor. PMID- 15364205 TI - Effects of 17beta-estradiol, nonylphenol, and bisphenol-A on developing Xenopus laevis embryos. AB - Many chemicals released into the environment have the capacity to disrupt the normal development of aquatic animals. We investigated the influence of nonylphenol (NP), bisphenol-A (BPA), and 17beta-estradiol (E2) on developing Xenopus laevis embryos, as a model animal in the aquatic environment. Embryos were exposed to eight different concentrations of NP, BPA or E2 between 3 and 96 h post-fertilization (p.f.). Short body length, microcephaly, flexure, edema, and abnormal gut coiling were induced by 20 microM NP, BPA or 10 microM E2 by 96 h p.f. To clarify sensitive stages to these compounds, embryos were exposed to chemicals for 45 or 48 h starting at different developmental stages and experiments were terminated 96 h p.f. BPA and NP induced abnormalities in developing X. laevis, though the sensitive stages of embryos to these chemicals are different, BPA affecting earlier stages and NP affecting at later stages. To analyze the functional mechanisms of BPA and NP in induction of morphological changes, we adapted a DNA array technology and identified 6 X. laevis genes, XIRG, alpha skeletal tropomyosin, cyclin G1, HGF, troponin C2, and ribosomal protein L9. These findings may provide important clues to elucidate common mechanisms underlying teratogenic effects of these chemicals. PMID- 15364206 TI - Effects of follicle-stimulating hormone and androgen on proliferation of cultured testicular germ cells of embryonic chickens. AB - A germ-Sertoli cell coculture model was established to study effects of follicle stimulating hormone (FSH) and testosterone (T) on testicular germ cell proliferation of the embryonic chickens. Germ and somatic cells were dispersed from 18-day-old embryonic testes and cultured in 96-well plates. Germ cells were characterized by expression of stem cell factor receptor c-kit. Germ cell proliferation was assessed by an increase in cell number and expression of proliferating cell nuclear antigen (PCNA). Results showed that the germ and Sertoli cells kept alive in serum-free McCoy's 5A medium supplemented with insulin, transferrin, and selenite (ITS medium). Germ cells adhered to the free surface of Sertoli cells that spread the filopodia and formed a monolayer in ITS medium. In the serum-containing medium, Sertoli cells displayed an increment with a flat squamous form and only a few very large germ cell masses were found in the free surface of Sertoli cells. Many germ cells showed apoptosis in the McCoy's 5A medium without ITS or serum. Only germ cells showed positive staining for c-kit in the coculture. Ovine FSH (0.25-1.0 IU/ml) significantly increased the number of germ cells, and PCNA-labeling index (P < 0.05). FSH also induced stronger c kit expression compared with the control. In the FSH-treated groups, germ cells were manifested distinct knob-like form. Similar stimulating effect was found in the germ cell number by T treatments (10(-7)-10(-6)M). Furthermore, FSH (0.5 IU/ml) combined with T significantly promoted higher testicular germ cell proliferation (P < 0.05) compared with either FSH or T alone, which indicated that interaction of FSH and T might be additive. The above results showed that the serum-free germ-Sertoli cell coculture model allowed evaluating hormonal regulation of testicular germ cell proliferation. FSH and T promoted testicular germ cell proliferation probably through indirect effects on Sertoli cells. PMID- 15364207 TI - Sequence and expression of cytochrome P450 aromatase and FTZ-F1 genes in the protandrous black porgy (Acanthopagrus schlegeli). AB - In this study, a cDNA encoding cytochrome P450 aromatase (P450arom) was cloned from black porgy Acanthopagrus schlegeli ovary. The deduced amino acid sequence had high homology with ovarian P450arom of other teleost fish. Moreover, we partially cloned two FTZ-F1 homologues (asff1a and asff1b) from black porgy. Comparative sequence analysis grouped asff1a and asff1b in NR5A2 and NR5A4 clades, respectively. Among the various tissues tested, P450arom mRNA was highly expressed in the ovary and weakly in the brain and testis, asff1a was expressed in brain, liver, intestine, kidney, testis, and ovary, asff1b was expressed in brain, kidney, testis, and ovary. The transcript levels of P450arom, asff1a, and asff1b were measured in the ovary and testis of 1+ -year-old, 2+ -year-old, and 5+ -year-old black porgy. The transcript level of P450arom in the ovary of 2+ year-old fish was significantly higher than those of 1+ -year-old and 5+ -year old fish. The results suggest that P450arom gene may be involved in the mechanism of natural sex change of protandrous black porgy. No change in ovarian expression of asff1a or asff1b was observed among different ages. These results suggest that up-regulation of the transcript levels of P450arom during the course of natural sex change of black porgy was not regulated via FTZ-F1. PMID- 15364208 TI - The role of sex ratio on spawning performance and on the free and conjugated sex steroids released into the water by common dentex (Dentex dentex) broodstock. AB - This paper reports the results of an experiment to determine whether sex ratio has any effect on the spawning efficiency of common dentex, Dentex dentex, broodstock. The paper also reports preliminary data on diurnal changes in water concentrations of steroids (measured on two separate occasions) in the same groups of fish. These latter observations were made to determine whether, using a non-invasive procedure, it was possible to relate hormonal changes in the fish to the "success of spawning." Mature fish were placed in broodstock tanks at three different sex ratios (Group A, 4 female:1 male; Group B, 1 female:1 male; and Group C, 1 female:4 male) and regularly monitored for egg production. At 18 days after the start of spawning, water samples were removed from all three tanks at 130 min intervals over a period of 24 h. At the end of spawning, water samples were removed at 4 h intervals. Free and conjugated steroids were extracted from the water and measured by radioimmunoassay. The highest number of eggs released, percentage of viable eggs, number of spawnings, spawning index, and relative fecundity were observed in Group B (1 female:1 male). The sex steroids testosterone (T), 11-ketotestosterone (11-KT), 17,20beta-P, and 17,20beta, 21 trihydroxy-pregn-4-en-3-one could be readily detected in water from all tanks. Concentrations of the conjugated forms (sulphate and glucuronide) were higher than those of the free form--with the exception of sulphated T and 11-KT. The concentrations of glucuronidated 17,20beta-P, sulphated 17,20beta-P, and free T appeared to be associated with the number of females in the tank and glucuronidated 11-KT concentrations with the number of males. In the first set of water samples (taken during the spawning period) there was also evidence for a cycle in steroid release with an acrophase during the afternoon and early evening, a few hours before the actual act of spawning. We were not able to show any association between steroid concentrations and success of spawning. However, since the data were derived from a "snapshot" of just a single day in what was a prolonged spawning period, this was perhaps not surprising. The results from the present study will help us to design experiments that are more able to answer this question. PMID- 15364209 TI - Species and fetal gender effects on the endocrinology of pregnancy in elephants. AB - Quantitative and temporal progestin profiles vary during gestation in the elephant, sometimes making it difficult to determine if a pregnancy is progressing normally. The aim of the present study was to determine if circulating progestin variability was related to species or fetal gender effects. A similar comparison also was conducted for secretory profiles of prolactin, relaxin, and cortisol. Overall mean progestin concentrations during gestation in Asian (n = 19) and African (n = 8) elephants were similar; however, the temporal profiles differed (P < 0.001). Concentrations were higher in African elephants during the first half of pregnancy, but then declined to levels below those observed in Asian elephants (P < 0.05). There also was a fetal gender effect in Asian, but not African elephants. Progestin concentrations were higher in Asian cows carrying male calves (n = 9) as compared to those carrying females (n = 10) (P < 0.001). Overall prolactin concentrations were higher in Asian than in African elephants between 8 and 15 months of gestation ( P< 0.001). There were no species differences in the secretory patterns of relaxin. Cortisol was relatively stable until the end of gestation when significant surges were observed, mainly between 8 and 11 days before parturition, and again on the day of birth. In sum, a comparison of progestin patterns between Asian and African elephants identified notable differences related to species and fetal gender. A role for cortisol in the initiation of parturition also was inferred from these data. From a practical standpoint, understanding the factors affecting gestational hormone characteristics and recognizing what the species differences are will help ensure that data used in diagnosing and monitoring elephant pregnancies are properly interpreted. PMID- 15364210 TI - Cloning and developmental expression of the cytochrome P450 aromatase gene (CYP19) in the European eel (Anguilla anguilla). AB - To characterize the involvement of the aromatase gene during the process of sex determination in the European eel (Anguilla anguilla), the expression of its gonadal form was determined during various developmental stages. The cloned cDNA from the European eel gonad (EeCYP19) contains an open reading frame of 1539 bp, encoding a deduced protein of 513 residues. The predicted amino acid sequence shows 97% identity with that of the Japanese eel, and 59-69% of identity with those of the CYP19 gonadal and brain forms of other teleost fish. Two potential initiation sites (ATG) were found downstream of the first ATG codon. A fluorescent-based method of real-time PCR was developed to quantify EeCYP19 expression. The expression levels of EeCYP19 in the gonads of adult males were approximately 12- and 30-fold lower than the levels in adult females and juvenile eels previously treated with E2, respectively. Expression of aromatase was found only in a single specimen in the control group. In contrast, no difference was found among sexes in the aromatase expression in the brain. Treatment with aromatase inhibitor (AI) of juvenile eel resulted in the total loss of aromatase expression in the gonads and brains. The results of this work revealed that AI treatment not only reduces the synthesis of estradiol, but reduces the expression levels of EeCYP19 as well. No evidence for the presence of a distinct extra gonadal (brain) form of aromatase in the European eel could be provided. PMID- 15364211 TI - Sequence analysis of cDNA encoding follicle-stimulating hormone and luteinizing hormone beta-subunits in the Mastomys (Praomys coucha). AB - To examine the molecular basis of efficient superovulation in the Mastomys (Praomys coucha), the cDNA sequences of the follicle-stimulating hormone (FSH) and luteinizing hormone (LH) beta-subunits were determined and compared with those of other mammals. FSHbeta and LHbeta cDNAs were 1606 and 513 bp long, respectively, from the 5'-ends to the putative polyA sites. The deduced sequences of the FSHbeta and LHbeta precursor proteins were 130 and 141 amino acids in length, respectively. The amino acid sequences of both mastomys hormone subunits showed overall similarity to those of other rodents. In particular, the N terminus of the FSHbeta precursor protein is of the MM-type, like those of mice and rats, which suggests that the MM-type is characteristic of the subfamily Murinae. As we reported earlier for the Mongolian gerbil, the use of hCG for superovulation of mastomys might need to be re-evaluated, due to the low homology between hCG and rodent LH sequences. PMID- 15364212 TI - The association of prior cytomegalovirus infection with neovascular age-related macular degeneration. AB - PURPOSE: To determine if prior exposure to pathogens associated with vascular disease, cytomegalovirus, Chlamydia pneumoniae, and Helicobacter pylori correlates with neovascular age-related macular degeneration (AMD). DESIGN: An experimental study. METHODS: SETTING: Institutional. Bascom Palmer Eye Institute, October 2001 to December 2002. PATIENT POPULATION: 150 patients (47 neovascular amd, 36 dry amd, and 67 non-amd controls) were included in the study. exclusion criteria included hiv infection, malignancy, recent acute illness requiring hospitalization within 6 months, or immunosuppressive illness. PROCEDURE: Serum samples were obtained for analysis of cytomegalovirus, chlamydia pneumoniae, and helicobacter pylori igg antibody titers by elisa. MAIN OUTCOME MEASURE: Comparison of the distribution of igg titers between patients with wet amd, dry amd, and controls. RESULTS: The average cytomegalovirus IgG titer was higher in patients with wet AMD versus controls (p = 0.02, Student t-test, two tailed) and patients with dry AMD (p = 0.06). Twenty-six (55%) of 47 subjects with wet AMD had high cytomegalovirus IgG titers compared with 14 (39%) of 36 patients with dry AMD (odds ratio [OR] = 2.23, 95% confidence interval [CI] = 0.77 to 6.44) and 23 (34%) of 67 control patients (OR = 2.49, 95% CI = 0.98 to 6.33). There was no major difference in the distribution of titers for Chlamydia pneumoniae IgG and Helicobacter pylori IgG in wet and dry AMD patients. Five of 47 patients with wet AMD (11%) had high antibody titers to all three pathogens, compared with only 1 of 36 patients with dry AMD (3%) (OR = 4.17, 95% CI = 0.46 to 37.36). CONCLUSIONS: There was a significant association of high cytomegalovirus IgG titer with neovascular AMD compared with dry AMD and control patients. Chronic infection with cytomegalovirus may be a novel risk factor for the progression from dry to neovascular AMD. PMID- 15364213 TI - Chemoreduction for retinoblastoma. Analysis of tumor control and risks for recurrence in 457 tumors. AB - PURPOSE: To evaluate retinoblastoma control following chemoreduction. DESIGN: Interventional case series. METHODS: Prospective. SETTING: Single center trial. PATIENT POPULATION: 457 retinoblastomas in 193 eyes of 125 patients. Nonrandomized, noncomparative study. INTERVENTION: All patients received intravenous vincristine, etoposide, and carboplatin,. The tumors were managed with chemoreduction alone (group W) or chemoreduction combined with thermotherapy (group X), cryotherapy (group Y), or both thermotherapy and cryotherapy (group Z). MAIN OUTCOME MEASURE: Tumor recurrence in each treatment group. RESULTS: Of 457 retinoblastomas, 63 (14%) were in group W, 256 (56%) in group X, 127 (28%) in group Y, and 11 (2%) in group Z. The tumor was located in the macula in 33 (52%) of group W, 109 (43%) of group X, 3 (2%) of group Y, and 9 (1%) of group Z. The mean tumor thickness at initial examination was 7 mm for group W, 4 mm for group X, 2 mm for group Y, and 3 mm for group Z. Using Kaplan-Meier estimates, recurrence of the individual retinoblastoma at 7 years was found in 45% of group W and 18% for combined groups X, Y, and Z. Risk factors predictive of tumor recurrence by multivariate analysis included macular tumor location for all groups and additionally female gender for group W and increasing tumor thickness for groups X, Y, and Z. CONCLUSIONS: Chemoreduction alone or combined with cryotherapy or thermotherapy is effective for treatment of retinoblastoma, but tumor recurrence rate is highest for those located in the macula and those with greater thickness. PMID- 15364214 TI - Clinical characteristics and outcomes of cytomegalovirus retinitis in persons without human immunodeficiency virus infection. AB - PURPOSE: To describe the characteristics and outcomes of patients with cytomegalovirus (CMV) retinitis in the absence of human immunodeficiency virus (HIV) infection. DESIGN: Retrospective cohort study. METHODS: Consecutive patients with CMV retinitis in the absence of HIV infection were identified at a university hospital. Demographic and clinical characteristics were noted at the time of CMV retinitis. Outcomes were determined retrospectively. Main outcome measures were rates of second eye involvement, vision loss, rhegmatogenous retinal detachment (RD), immune recovery uveitis, progression of retinitis, and mortality. RESULTS: The clinical characteristics of CMV retinitis in 18 patients (30 eyes) without HIV infection diagnosed between January 1, 1984, and April 13, 2003, were similar to those of patients with HIV infection. The incidences of visual loss to the levels of 20/50 or worse and of 20/200 or worse were 17% per eye-year and 14% per eye-year, respectively. The observed incidence of RD was 3.7% per eye-year, and the mortality rate was 23% per person-year. Following reduction of immunosuppression, 10 patients (56%) who discontinued anti-CMV therapy remained free of retinitis progression. The incidence of immune recovery uveitis was 13% per person-year. CONCLUSIONS: In our series, CMV retinitis in patients without HIV infection had a clinical course similar to that in patients with AIDS treated with highly active antiretroviral therapy (HAART), except the incidence of RD was lower for patients without AIDS. A substantial number of patients no longer required long-term anti-CMV therapy after adjustment of immunomodulatory therapy. PMID- 15364215 TI - Visual field attention is reduced by concomitant hands-free conversation on a cellular telephone. AB - PURPOSE: To quantify the central attention-diverting effect of hands-free cellular phone conversation on visual field awareness. DESIGN: Experimental study. METHODS: Twenty male and 21 female healthy participants performed a pretest and baseline Esterman visual field examinations with the Humphrey Systems Visual Field Analyzer II. During the consequent third examination, each participant engaged in a hands-free conversation using a cellular phone. The conversation was the same for all participants. Visual field performance parameters were compared between the second (baseline) examination, and the third (test) examination for each eye. RESULTS: During phone conversation, missed points increased from mean 1.0 +/- 1.5 to 2.6 +/- 3.4 (P < or =.001) in the right eye and from 1.1 +/- 1.53 to 3.0 +/- 3.4 (P <.001) in the left eye. Fixation loss increased from mean 7.8% to 27.4% (P <.0001) and from 7.2% to 34.8% (P <.0001) for the right and left eyes, respectively. Test duration increased by a mean of 0.28 seconds (15%) per stimulus (P <.0001). Approximately half of missed points were inside the central 30 degrees. There was no significant difference in the performance of male and female participants. CONCLUSION: We describe a new model for the quantification of the attention-diverting effect of cellular-phone conversation on the visual field. In the current study, cellular hands-free conversation caused some subjects to miss significantly more points, react slower to each stimulus, and perform with reduced precision. Legislative restrictions on concomitant cellular-phone conversation and driving may need to be based on individual performance rather than a general ban on cellular phone usage. PMID- 15364216 TI - Optical coherence tomography and confocal scanning laser tomography for assessment of macular edema. AB - PURPOSE: To compare optical coherence tomography (OCT) and confocal scanning laser tomography (cSLT) for quantitative retinal thickness mapping of the macula and their ability to detect macular edema. DESIGN: Prospective, comparative, clinical observational study. METHODS: The study population of 138 eyes (97 patients) was divided into a study group consisting of 45 (32.6%) eyes with macular edema and a control group consisting of 93 (67.4%) eyes without macular edema. All patients underwent OCT and cSLT of the macula. Retinal thickness measurements obtained by OCT were compared with signal width and edema index, determined by cSLT. RESULTS: The OCT measurements and cSLT edema index were significantly (P <.001) correlated with each other. Correlation coefficients decreased (P <.001) with increasing diameter of the measurement circle. In the macular edema group, correlation coefficients were significantly (P <.001) higher than in the control group. To separate the study and control groups, receiver operator characteristic curves covered a larger area for OCT measurements than for cSLT measurements. Retinal thickness measurements and edema index correlate with visual acuity (correlation coefficient r = -.653 for OCT, r = -.608 for cSLT; P <.001). CONCLUSIONS: Macular edema can be quantitatively mapped by OCT and cSLT. The retinal thickness and edema index measurements correlate with visual acuity. The fast and standard examination modes of OCT give similar measurements. Both OCT and cSLT can differentiate between eyes with and without macular edema, with OCT showing a higher predictive value. PMID- 15364217 TI - Development of an instrument to assess vision-related quality of life in young children. AB - PURPOSE: Quality of life (QOL) instruments are increasingly utilized in ophthalmological research. Measuring vision-related QOL in young children is complicated by constantly evolving abilities related to normal growth and development. Our aim was to develop vision-related QOL instruments for children in different age ranges <==7 years, and to provide initial validation of these instruments. DESIGN: Cross-sectional study. METHODS: SETTING: Multicenter. PATIENTS: 773 pediatric patients (age <==7 years) with a wide range of ophthalmological diagnoses. PROCEDURE: Questionnaire. A 61-item prototype instrument with a wide variety of items was applied to 403 consecutive patients. The usefulness of items was evaluated as a function of age in order to derive two age-group specific instruments and to find the age limit dividing the age groups. Thus, age-specific versions of a Children's Visual Function Questionnaire (CVFQ) were defined for ages <3 years and >/=3 years, and applied to a convenience sample of patients. OUTCOME: Subscale scores. Factor analysis helped identify underlying dimensions of the data, and corresponding subscales were defined. Validation was provided by examining the internal consistency reliability and by exploring the associations between scores and clinical characteristics. RESULTS: Subscales for General health, General vision, Competence, Personality, Family impact, and Treatment were defined, with internal consistency reliabilities ranging from 0.60 to 0.86. The association between subscales scores and age was weak, whereas strong correlations were found with the level of visual impairment and type of visual diagnosis. CONCLUSION: The CVFQ assesses the impact of visual impairment on children and their families, and is expected to become a useful tool for the pediatric vision research community. PMID- 15364218 TI - Uveitis in Behcet disease: an analysis of 880 patients. AB - PURPOSE: We aimed to describe the demographic and clinical features, ocular manifestations, complications, visual prognosis, and treatment in a large population of Turkish patients with Behcet uveitis. We also aimed to compare visual prognosis between male and female sex and between patients who presented before and after 1990. DESIGN: Observational case series. METHODS: A retrospective study of 880 consecutive patients (1,567 eyes) with Behcet uveitis seen at the Uveitis Service, Department of Ophthalmology, Istanbul Faculty of Medicine, Istanbul University, from 1980 to 1998. All patients met the classification criteria of the International Study Group for Behcet's Disease. Information on the patient's sex, age at onset of uveitis, ocular features, ocular complications, visual acuity, and systemic treatment was collected. RESULTS: Five hundred ninety-nine patients (68%) were male and 281 (32%) were female. The mean age at onset of uveitis was 28.5 years in male and 30 years in female patients. Ocular involvement was bilateral in 78.1% and unilateral in 21.9% of the patients. Panuveitis was the most common form in both sexes. Fundus lesions as well as sight-threatening complications were more common in males. At the beginning of the follow-up, potential visual acuity was 0.1 or less in 30.9% of eyes in males and 24.2% of eyes in females. Kaplan-Meier survival analysis estimated the risks of losing useful vision (>0.1) at 5 and 10 years for males and females as 21% vs 10% and 30% vs 17%, respectively. Male patients who presented in the 1990s had a significantly lower risk of losing vision compared with male patients who presented in the 1980s. CONCLUSION: Behcet uveitis starts frequently around the end of the third decade and has a male predominance. The disease is more severe and the risk of losing useful vision is higher in males than in females. However, this risk has been significantly reduced in the 1990s. PMID- 15364219 TI - Ocular surface changes with applanation contact lens and coupling fluid use after argon laser photocoagulation in noninsulin-dependent diabetes mellitus. AB - PURPOSE: To describe the effect of coupling solutions used during laser photocoagulation on the ocular surface of patients with noninsulin-dependent diabetes mellitus (NIDDM). DESIGN: A prospective case-controlled study. METHODS: Ninety-two eyes of 46 NIDDM patients with clinically significant macular edema, poor metabolic control of diabetes, and peripheral neuropathy and 100 eyes of 50 normal control subjects were studied. The patients' eyes were assigned to argon green focal/grid laser photocoagulation using an applanation contact lens and one of the coupling fluids; 2% methocel, Thilo-Tears Gel, 1.4% sodium hyaluronate, or 0.9% simple saline. The control subjects received time-matched three-mirror contact lens fundus examinations. All subjects underwent corneal sensitivity measurements, Schirmer test, tear film breakup time, and corneal fluorescein staining before as well as 3 and 8 days after the laser procedures and contact lens examinations. Patients with corneal problems persisting after 8 days were followed longer. RESULTS: Diabetic eyes assigned to 2% methocel and 1.4% sodium hyaluronate had significantly lower mean corneal sensitivities and break-up time values as well as significantly higher mean fluorescein staining scores at all examination points after laser photocoagulation. All diabetic eyes with aqueous deficiency assigned to 2% methocel and 1.4% sodium hyaluronate developed delayed corneal epithelial healing. CONCLUSION: The use of viscous coupling solutions during applanation contact lens-aided laser procedures may be detrimental for the corneal epithelium in poorly controlled NIDDM patients with peripheral neuropathy and coexisting aqueous deficiency. PMID- 15364220 TI - Comparison of the nocturnal effects of once-daily timolol and latanoprost on intraocular pressure. AB - PURPOSE: To compare the nocturnal effects of once-daily timolol and latanoprost on intraocular pressure (IOP) in patients with ocular hypertension or early glaucomatous changes. DESIGN: Prospective, open-label, experimental study with crossover design. METHODS: Eighteen patients with ocular hypertension or early glaucomatous changes (aged 41 to 79 years) each received topical treatments with timolol (0.5% Timoptic-XE), latanoprost (0.005% Xalatan), and no IOP-lowering medication, for at least 4 weeks. Timolol was given once in the morning upon awakening and latanoprost once in the evening at bedtime. At the end of each treatment period, the patient was housed in a sleep laboratory for 24 hours and IOP was measured every 2 hours using a pneumatonometer. Measurements were taken sitting and supine during the 16-hour diurnal/wake period and only supine during the 8-hour nocturnal/sleep period. Mean diurnal and nocturnal IOP levels were compared among the treatments with timolol, latanoprost, and no medication. RESULTS: In the diurnal period, the mean IOP under the timolol or the latanoprost treatment was significantly less than the mean IOP under no medication in both the sitting and the supine positions. There was no statistical difference between the timolol and latanoprost treatments. In the nocturnal period, supine IOP with timolol treatment was not different from the supine IOP with no medication but was significantly higher than supine IOP with the latanoprost treatment. CONCLUSION: Although both once-daily timolol and latanoprost were effective in lowering IOP during the diurnal period, only latanoprost reduced IOP during the nocturnal period. PMID- 15364221 TI - Inflammatory orbital pseudotumor with extension beyond the orbit. AB - PURPOSE: To summarize the clinical profile and response to treatment of 4 biopsy proven cases of inflammatory orbital pseudotumor extending beyond the orbit. DESIGN: Retrospective observational case series. METHODS: Four patients' charts were retrospectively reviewed. There were three men with extraorbital extension (two intracranial; one maxillary antrum) and one woman with intracranial disease that extended into the orbit. The men were 40, 41, and 60 years old; the woman was 73 years old. RESULTS: Two men with orbital myositis and mild discomfort, initially treated with corticosteroids, had asymptomatic intracranial disease 9 and 12 months after initial presentation. A third man had extension into the maxillary antrum after initial symptoms of painless diplopia. The woman had intraorbital disease with minimal discomfort (dacryoadenitis and myositis) 5 years after presenting with intracranial disease in the Meckel cave that subsequently became bilateral. Histopathologic examination in all cases showed nonspecific inflammation without evidence of vasculitis or granulomas. CONCLUSIONS: Unlike typical cases of nonspecific orbital inflammation, two of these four cases did not have pain as a prominent feature. Neuroimaging was essential in diagnosing asymptomatic extraorbital disease. Surgery has a prominent role in confirming this diagnosis, primarily by helping to rule out other diseases, such as those with granulomatous inflammation or vasculitis. Additional therapy was usually not required postoperatively. PMID- 15364222 TI - Chemokines and gelatinases in the aqueous humor of patients with active uveitis. AB - PURPOSE: To investigate the involvement of the chemokines CXCL10/IP-10, CXCL11/I TAC, CXCL8/1L-8, CXCL6/GCP-2, CCL3/MIP-1alpha, and CCL18/PARC, and gelatinases A and B in uveitis. DESIGN: Prospective, experimental, case-control study. METHODS: Aqueous humor samples from 30 patients with active uveitis, and 14 control patients and paired serum samples were assayed for chemokines with specific enzyme-linked immunosorbent assays (ELISAs) and for gelatinase levels by quantitative zymography. RESULTS: In control AH, none of the chemokines was detected. Gelatinase A was detected in all samples, and gelatinase B was detected in only one sample. In patients with uveitis, IP-10 was detected in all AH samples, whereas I-TAC, IL-8, GCP-2, MIP-1alpha, and PARC were detected in three, 16, six, two, and 12 samples, respectively. IP-10 levels were significantly higher in AH samples than those of serum (P =.006). Gelatinase A was detected in 29 AH samples and gelatinase B was detected in 26 samples. Gelatinase A levels were significantly higher in AH samples from patients than those of controls (P <.0001). In 11 AH samples, gelatinase B was detected in complex with lipocalin (NGAL). Disease activity correlated significantly with the levels of IP-10 (r =.627; P <.0001), gelatinase A (r =.508; P =.002), gelatinase B (r =.685; P <.0001), and NGAL-gelatinase B complex (r =.595; P <.0001). CONCLUSIONS: These data suggest a pathogenic role of the T lymphocyte chemoattractant IP-10 and gelatinases in the recruitment and activity of T cells into the eye in patients with uveitis and in the pathogenesis of uveitis. PMID- 15364223 TI - Ophthalmic imaging by spectral optical coherence tomography. AB - PURPOSE: To demonstrate two novel ophthalmic imaging techniques based on fast Spectral Optical Coherence Tomography (SOCT). DESIGN: Prospective observation case report. METHODS: SETTING: Research laboratory. STUDY POPULATION: A normal human subject. MAIN OUTCOME MEASURES: Correlation of an optical coherence retinal tomogram with known retinal anatomy, reduction of eye exposure to the probing light beam during Optical Coherence Tomography (OCT) examination. RESULTS: High resolution (3- x 20-microm) cross-sectional 2-dimensional images of the human retina in vivo have been obtained with only 0.1-second total illumination time (for 1024 A-scans), and approximately 200- microW eye exposure. Details of the optic disk in vivo have been visualized at a rate of eight frames per second, which is sufficient to provide real time analysis. The 3-dimensional images of the optic nerve correspond to the images obtained from a fundus camera. CONCLUSIONS: Because of its short acquisition time and high sensitivity, SOCT uses a 100 times lower exposure/A-scan to obtain images of comparable quality to those obtained by the commercial instrument based on traditional time-domain OCT. Spectral Optical Coherence Tomography therefore provides a much wider safety margin than the traditional method and allows relaxation of the current safety precautions forbidding more than 10 minutes/day scanning of the same location of a retina. As a result of the high speed offered by the new technique, the 3 dimensional tomograms, which allow a surgeon access to a comprehensive and detailed view of relevant pathologies, are obtained in a much shorter, clinically more reasonable time. PMID- 15364224 TI - Late porous polyethylene implant exposure after motility coupling post placement. AB - PURPOSE: To report the probable association of motility coupling post placement and late porous polyethylene implant exposure. DESIGN: Retrospective, observational case series. METHODS: This was a retrospective analysis of 27 patients who had primary porous polyethylene orbital implantation from February 1999 to November 2000. Data on demographics, previous surgery, ocular diagnosis, type of surgery, size of the implant, and motility coupling post placement were collected. Complications of porous polyethylene implants and implant exposure were documented. RESULTS: Among the 27 patients, 18 eyes (66.7%) received motility coupling post insertion after primary porous polyethylene implantation. Implant exposure occurred in six (33.3%) of the 18 eyes with motility coupling post insertion. None of the eyes without insertion had implant exposure. The mean interval between porous polyethylene implantation and motility coupling post placement for the implant exposure group (6 of 18) was 6.5 +/- 0.4 months, which was not statistically significant compared with 7.2 +/- 0.6 months in the nonexposure group (12 of 18). For these 6 cases of implant exposure, the mean interval between implantation and implant exposure was 24.2 +/- 11.8 months, and the mean interval between pegging and exposure was 17.6 +/- 11.7 months. Among these 6 patients, 4 underwent removal of exposed porous polyethylene implants and reimplantation of hydroxyapatite implant or dermis fat reconstruction. CONCLUSIONS: We found a trend (P =.07) of increasing risk of porous polyethylene implant exposure with motility coupling post placement. Although the pegging group did not show a statistically significant higher rate of exposure compared with the nonpegging group, we believe that more care was needed when performing motility coupling post placement. In addition, longer postoperative follow-up is needed after insertion of a motility coupling post. PMID- 15364225 TI - Porous polyethylene orbital implant in the pediatric population. AB - PURPOSE: To determine the incidence of complications of primary insertion of porous polyethylene orbital implant in the pediatric population. DESIGN: Interventional case series. METHODS: Prospective nonrandomized case series of 36 eyes of 36 patients under age 15 years who underwent primary placement of an anteriorly wrapped spherical porous polyethylene orbital implant from March 1998 to August 2002, with at least 17 months of follow-up. RESULTS: The mean age at the time of surgery was 4.6 years. The histopathologic diagnoses after enucleation included intraocular tumor in 22 patients, phthisis bulbi in eight patients, microphthalmos in three patients, Coats disease in two patients, and ruptured traumatic globe in one patient. Twelve patients (33%) had prior ocular surgery. At the time of enucleation, all patients underwent primary placement of spherical porous polyethylene orbital implant anteriorly wrapped with homologous sclera in 30 patients (83%) and autologous sclera in six patients (17%). The spherical implant size was 16 mm in one patient (3%), 18 mm in 10 patients ( 28%), and 20 mm in 25 patients (69%). The prosthesis was fitted after a mean interval of 5 weeks. After a mean follow-up of 44 months (range, 17 to 68 months), there was one case of pyogenic granuloma (3%) and one case of implant exposure (3%). There were no cases of implant extrusion, superior sulcus syndrome, orbital cellulitis, or significant inflammatory response. No porous polyethylene orbital implant was drilled for peg placement. CONCLUSIONS: Anteriorly wrapped primary porous polyethylene orbital implant in the pediatric population appears to be well tolerated with few complications. PMID- 15364226 TI - Donor site morbidity in free tarsal grafts. AB - PURPOSE: To determine if harvesting free tarsal grafts from the upper eyelids is associated with significant morbidity. DESIGN: Prospective case series. METHODS: Consecutive patients managed with free tarsal grafts by three of the authors (D.S., D.G., R.G.) over a 4-year period. RESULTS: Ninety-one eyelids in 75 patients were operated on using the free tarsal graft for various eyelid pathologies. Mean follow-up period was 14.83 +/- 8.45 months (range, 4 to 42 months). Contour peaking in donor upper eyelid appeared in two eyelids (2.2%), lash ptosis in four eyelids (4.4%) of three patients, and donor site infection in one eyelid (1.1%). Four patients (5.3%) complained of a transient ocular discomfort. There were no cases of postoperative changes on the eyelid height, entropion, or keratopathy in the donor site eye. Most of the donor site complications were mild or transient and required no further surgery. In three patients who underwent subsequent surgery to treat preexisting ptosis or retraction of the donor eyelid, the results were less predictable and further procedures were required. CONCLUSION: Free tarsal grafts harvested from the upper eyelids, with preservation of at least 4 mm of the tarsal plate inferiorly, do not lead to significant donor site morbidity. However, subsequent procedures to alter the position of the donor eyelid may be less predictable. PMID- 15364227 TI - Prognostic factors influencing visual outcome of photodynamic therapy for subfoveal choroidal neovascularization in pathologic myopia. AB - PURPOSE: To examine the influence of age, lesion size, degree of myopia, and baseline visual acuity on the visual outcome of patients with pathologic myopia and choroidal neovascularization (CNV) who received photodynamic therapy (PDT) with verteporfin. DESIGN: Retrospective, noncomparative consecutive case series. METHODS: Forty-three eyes of 41 patients were treated in a two-year time span; 36 eyes of 36 patients who had received PDT for CNV due to pathologic myopia were examined for the above-mentioned factors 24 months after first treatment. All patients had been treated according to the Verteporfin in Photodynamic Therapy (VIP) study criteria. Patients were examined in two- to three-month intervals with Snellen visual acuity, biomicroscopy, and fluorescein angiography. RESULTS: Baseline visual acuity and age were both prognostic factors for visual outcome (P =.0097, P =.0055). Lesion size (greatest linear dimension) at baseline, refractive error, or the number of treatments had no influence on the outcome. CONCLUSION: Age and baseline visual acuity have an effect on visual outcome in patients receiving PDT due to CNV secondary to pathologic myopia. Younger patients and patients with higher baseline visual acuity had a better treatment outcome. PMID- 15364228 TI - Autosomal dominant congenital nystagmus is not linked to 6p12, 7p11, and 15q11 in a German family. AB - PURPOSE: Congenital nystagmus (CN) is an eye-movement disorder that usually starts within the first months of life. Autosomal dominant, autosomal recessive, and X-chromosomal pedigree patterns are observed. Causative genes are yet unknown. Several loci were implicated to contain disease-relevant genes for autosomal dominant CN (AD CN). AD CN cosegregated with a balanced translocation of 7;15 in a family. In a large black pedigree linkage was demonstrated to 6p12. DESIGN: In this study, we describe a large German family with AD congenital nystagmus. Linkage of AD in this family was tested with previously implicated loci. METHODS: Affected family members and unaffected members underwent genetic analysis. Key family members underwent ophthalmologic testing and oculography. RESULTS: No linkage of AD CN to the implicated loci on 6p12, and 7p11, and 15q11 was found in this study. CONCLUSION: In the presented pedigree genes on 15q11, and on the assumption of full penetrance, 6p12 and 7p11 are not involved in the development of AD congenital nystagmus. PMID- 15364229 TI - A randomized, double-masked, placebo-controlled, multicenter comparison of loteprednol etabonate ophthalmic suspension, 0.5%, and placebo for treatment of keratoconjunctivitis sicca in patients with delayed tear clearance. AB - PURPOSE: To evaluate loteprednol etabonate ophthalmic 0.5% suspension, versus placebo for treatment of the inflammatory component of keratoconjunctivitis sicca in patients with delayed tear clearance. DESIGN: Randomized, double-masked, placebo-controlled clinical trial. METHODS: Sixty-four patients with keratoconjunctivitis sicca and delayed tear clearance were randomly assigned to receive either loteprednol or vehicle 4 times a day for 4 weeks. Patients were evaluated at weeks 2 and 4 of treatment and 2 weeks after treatment was discontinued. Symptoms were scored using a visual analog scale (VAS) of 1 to 100. Corneal fluorescein staining was scored 0 to 4 in five areas. Conjunctival injection was graded 0 to 3 in the inferior bulbar, nasal bulbar, and inferior tarsal areas. Lid margin injection was graded 0 to 3. Safety was assessed by funduscopy, lens examination, biomicroscopy, visual acuity, and Goldmann tonometry, and by monitoring adverse events and changes in symptoms. RESULTS: In subsets of patients with at least moderate clinical inflammation, there was a significant difference between the loteprednol-treated group and vehicle-treated group after 2 weeks of therapy. The differences did not reach statistical significance at 4 weeks, although the loteprednol-treated patients retained their improvement compared with the vehicle-treated group. Safety evaluations showed both treatments to be well tolerated and similar in the frequency and type of adverse event reported. CONCLUSION: The use of topical loteprednol etabonate 0.5% 4 times a day may be beneficial in patients who have keratoconjunctivitis sicca with at least a moderate inflammatory component. PMID- 15364230 TI - Risk assessment in the management of patients with ocular hypertension. AB - PURPOSE: To develop a model for estimating the global risk of disease progression in patients with ocular hypertension and to calculate the "number-needed-to treat" (NNT) to prevent progression to blindness as an aid to practitioners in clinical decision making. DESIGN: Development of a mathematical model for estimating risk of glaucoma progression. METHODS: Population-based studies of patients with ocular hypertension and glaucoma were reviewed by a panel of glaucoma specialists. Measures of disease progression risks derived from three long-term studies and assumptions based on the available data were used to estimate the risk of progression from ocular hypertension to glaucoma and glaucoma to unilateral blindness for untreated and treated patients over a 15 year period. Using these estimates, the NNT (1/absolute risk reduction on treatment) to prevent unilateral blindness in one patient with ocular hypertension was calculated. RESULTS: In untreated patients, the estimated risk of progression from ocular hypertension to unilateral blindness was 1.5% to 10.5% and in treated patients, the estimated risk of progression was 0.3% to 2.4% over 15 years. From these estimates, between 12 and 83 patients with ocular hypertension will require treatment to prevent one patient from progressing to unilateral blindness over a 15-year period. CONCLUSION: Global risk assessment that incorporates all available data plays a vital role in managing patients with ocular hypertension. A more precise understanding of long-term vision loss should be factored into decisions pertaining to the initiation of glaucoma therapy. Undoubtedly, these estimates will evolve and change with the availability of new population-based epidemiologic information and improvements in multivariable model testing. PMID- 15364231 TI - Infectious agents and ARMD: a connection? PMID- 15364232 TI - What is the best treatment for retinoblastoma? PMID- 15364233 TI - Herpes simplex virus keratitis in children. AB - PURPOSE: To describe the spectrum and recurrence of herpes simplex virus (HSV) keratitis in children and adolescents. DESIGN: Retrospective cohort study. METHODS: Twenty-three patients younger than age 16 years were diagnosed with HSV keratitis at one institution. RESULTS: All children presented with dendritic or punctate epithelial keratitis, and stromal keratitis occurred concurrently with epithelial keratitis in 14 patients (61%). Six patients (26%) had bilateral HSV keratitis. Eleven patients (48%) developed recurrent HSV keratitis at a median of 15 months after the first documented episode. Amblyopia occurred in three children. CONCLUSION: Children with herpetic keratitis may have bilateral ocular involvement and are at risk for recurrent keratitis and amblyopia. PMID- 15364234 TI - Immunopathology of progressive subretinal fibrosis: a variant of sympathetic ophthalmia. AB - PURPOSE: We report the immunopathology of progressive subretinal fibrosis, a variant of sympathetic ophthalmia. DESIGN: Brief case report. METHOD: Review of medical record and immunopathology of a chorioretinal biopsy of a 40-year-old man who presented with left total retinal detachment (RD) following multiple vitrectomies in the right eye for RD. Small peripherally retinal holes, snow banks, and inflammatory nodules along the ora serrata were observed during left vitrectomy. Both eyes deteriorated rapidly to blindness with progressive subretinal fibrosis and inflammation over 3 months, and the patient was unresponsive to systemic prednisolone and azathioprine. RESULT: Immunopathology of the chorioretinal biopsy of the right eye revealed aggregates of CD20+ B cells, surrounded by CD3+ T cells. CD68+ macrophages were scattered throughout. CONCLUSION: The rapid clinical course described in this case may be related to the unique immunopathology of rapidly forming peudogerminal centers in the choroids and retina. PMID- 15364235 TI - Intraobserver variability in optical coherence tomography. AB - PURPOSE: To assess the intraobserver variability of optical coherence tomography (OCT) in patients with clinically stable maculas. DESIGN: Retrospective, observational case series using a diagnostic instrument. METHODS: Retrospective study. setting. Private retina practice. patient population. Twenty-two eyes of 19 patients, each studied twice within 4 months. observation procedure. Optical coherence tomograph measurements of the macula obtained by two photographers. main outcome measures. Ordinary least products (OLP) and Bland-Altman analysis of OCT data. RESULTS: No fixed or proportional bias was detected in foveal zone thickness and total macular volume between measurements of either of the OCT operators. The coefficient of repeatability for foveal zone thickness was 37.0 microm for photographer 1 and 34.8 microm for photographer 2. The coefficient of repeatability for total macular volume was 0.29 mm(3) for photographer 1 and 0.10 mm(3) for photographer 2. CONCLUSION: Foveal zone thickness and total macular volume measurements show low intraobserver variability when analyzed by OLP and Bland-Altman techniques. Bounds are given for foveal zone thickness and total macular volume for which longitudinal OCT measurements by the same operator can be considered to reflect natural history or intervention effects rather than intraobserver variability. PMID- 15364236 TI - Optical coherence tomography of bilateral posterior microphthalmos with papillomacular fold and novel features of retinoschisis and dialysis. AB - PURPOSE: To report a case of retinoschisis and dialysis associated with bilateral posterior microphthalmos and papillomacular fold. DESIGN: Observational case series. METHODS: Complete ophthalmologic examination of three of five siblings presenting with bilateral posterior microphthalmos and papillomacular fold. Optical coherence tomography (OCT) data are presented to confirm the abnormal anatomy. RESULTS: All subjects have bilateral elevated horizontal papillomacular retinal fold with cystoid macular edema and shallow subretinal fluid. Optical coherence tomography was consistent with our examinations. One subject, a 13-year old Hispanic, initially presented with retinoschisis in the superotemporal quadrant of the left retina that developed 9 years later into a retinal dialysis without subretinal fluid. The right eye of this same patient developed retinoschisis in the far superotemporal retinal periphery during 9 years of observation. CONCLUSION: Retinoschisis and dialysis may occur in patients with posterior microphthalmos with papillomacular fold. Optical coherence tomography may be helpful in assessing these patients. PMID- 15364237 TI - Prevalence of factor XIII Val34Leu polymorphism in patients affected by spontaneous subconjunctival hemorrhage. AB - PURPOSE: To verify the prevalence of Val34Leu polymorphism in factor XIII A-chain gene (FXIII Val34Leu) in patients with spontaneous subconjunctival hemorrhage (SCH). DESIGN: Nonrandomized case-control study. METHODS: One hundred seven white patients suffering from one or more episodes of idiopathic SCH and 107 healthy subjects were matched for age and gender, and genotyped for FXIII Val34Leu. Anamnestic, ophthalmologic, cardiovascular, and serologic examinations were performed. RESULTS: Frequency of FXIII mutated allele (Leu34) was significantly higher in SCH patients than in controls. Computing together heterozygotes (Val/Leu) and homozygotes (Leu/Leu), genotype distribution was statistically different. In a conditional logistic regression model, the comparison of the three separated genotypes, performed among 25 patients with recurrent idiopathic SCHs and controls, gave significant differences for both Val/Leu and Leu/Leu variables. CONCLUSION: Both homozygosity and heterozygosity for FXIII Val34Leu predispose to idiopathic SCH, emphasizing the role of Leu34 allele as inherited risk factor for spontaneous, especially recurrent, SCHs. PMID- 15364238 TI - Acquired, isolated third nerve palsies in infants with cerebrovascular malformations. AB - PURPOSE: To report two infants with acquired, isolated third nerve palsies attributable to intracranial cerebrovascular malformations. DESIGN: Observational case report. METHODS: Two patients are described. Each was examined in a university-based pediatric ophthalmology and neuro-ophthalmology practice. RESULTS: An 8-month-old child presented with a pupil-involving partial left third nerve palsy because of a partially thrombosed fusiform aneurysm of the left internal carotid artery. A 3-month-old infant developed a right third nerve palsy from a giant arteriovenous fistula arising from an M2 branch of the right middle cerebral artery. CONCLUSION: Patients younger than 8 months and 3 months with acquired, isolated third nerve palsies resulting from intracranial cerebrovascular malformations could not be found in a MEDLINE search. In conclusion, at even this young age, acquired, isolated third nerve palsies may be the initial manifestation of an intracranial aneurysm or fistula. Magnetic resonance imaging (MRI) and MRI-angiography were adequate for detecting these processes. PMID- 15364239 TI - Oxygen permeability of amniotic membrane and actual tear oxygen tension beneath amniotic membrane patch. AB - PURPOSE: To investigate the oxygen permeability (Dk) and oxygen transmissibility (Dk/t) of amniotic membrane (AM), and tear oxygen tension beneath AM in rabbits. DESIGN: Experimental study. METHOD: The water content of AM was measured to calculate Dk and Dk/t, and compared with those of a therapeutic soft contact lens (TSCL). The tear oxygen tension beneath AM and TSCL was also measured in four male albino rabbits. RESULTS: The average water content of AM was significantly higher than that of TSCL (96.8 +/- 0.8% (mean +/- SD) vs 78%, P =.0006), giving rise to a significantly higher Dk and Dk/t in AM as compared with TSCL (142.8 +/- 4.7 vs 65.3, respectively, P =.0012, and 92.9 +/- 3.0 vs 29.7, respectively, P =.0008). The average tear oxygen tension under AM was also higher than TSCL (94.9 +/- 2.9 vs 59.1 +/- 4.9 mm Hg, P =.0006). CONCLUSIONS: These findings support the rationale of using AM as a superior bandage in treating persistent corneal epithelial defects or ulcers. PMID- 15364240 TI - Vitrectomy for a persisting macular fold in a case of resolved hypotony maculopathy. AB - PURPOSE: To describe a patient with resolved hypotony maculopathy with a persistent retinal fold (despite normalization of intraocular pressure [IOP]) who underwent successful surgical intervention by vitrectomy, internal limiting membrane peel, and gas tamponade. DESIGN: Interventional case report. METHODS: A 55-year-old man with a hypotony-induced macular retinal fold that did not improve following normalization of IOP underwent vitrectomy, internal limiting membrane peeling, and gas injection. Optical coherence tomography scans were performed both before and after surgery. RESULTS: Best-corrected visual acuity (BCVA) improved from 6/60 preoperatively to 6/9, with improvement in distortion. On repeat optical coherence tomography examination, the macular retinal fold had resolved. CONCLUSION: Vitrectomy, internal limiting membrane peeling and gas tamponade may be useful for cases of resolved hypotony maculopathy complicated by a persistent macular fold after normalization of IOP. PMID- 15364241 TI - Pseudohypopyon after intravitreal triamcinolone acetonide injection for cystoid macular edema. AB - PURPOSE: To report pseudohypopyon after intravitreal triamcinolone acetonide injection for cystoid macular edema. DESIGN: Retrospective, noncomparative, consecutive case series. METHODS: Records were reviewed of all patients who developed pseudohypopyon after intravitreal triamcinolone acetonide injection at Bascom Palmer Eye Institute between January 1, 2002 and February 1, 2004. RESULTS: A total of 828 intravitreal triamcinolone acetonide injections were administered to 686 patients during the study period. A pseudohypopyon (fine white crystalline opacities in the inferior anterior chamber angle) and suspended white crystalline opacities in the aqueous humor developed after 7 of the 828 injections (0.8%); all pseudohypopyons occurred within 3 days of injection and resolved completely within 2 weeks. None of the 686 patients developed clinically suspected infectious endophthalmitis. CONCLUSIONS: A transient pseudohypopyon may occur in the early postinjection period after intravitreal triamcinolone acetonide injection. Unless progressive intraocular inflammation occurs, close observational management is indicated. PMID- 15364242 TI - Trypan blue induces apoptosis in human retinal pigment epithelial cells. AB - PURPOSE: To examine whether trypan blue dye induces apoptosis in human retinal pigment epithelium cells. DESIGN: Laboratory investigation. METHODS: Pure cultures of human retinal pigment epithelium cells were isolated. The cells were incubated with different concentrations of trypan blue (0.5%, 0.10%, and 0.05%) for either 5 or 30 minutes. The rate of retinal pigment epithelium cell apoptosis was assessed with Annexin V-PE staining and flow cytometry. RESULTS: Trypan blue induced a statistically significant amount of apoptosis in retinal pigment epithelium cells at all the concentrations (0.5%, 0.10%, and 0.05%) (P <.05). The increase in incubation time (from 5 to 30 minutes) led to an increase in the number of apoptotic retinal pigment epithelium cells. CONCLUSION: The incubation of retinal pigment epithelium cells with trypan blue increased the number of apoptotic retinal pigment epithelium cells in vitro. Our results suggest that decisions regarding the intravitreal application of trypan blue dye need to be made with caution. PMID- 15364243 TI - Eyelid bleeding and atypical amyloidosis. AB - PURPOSE: To describe a case of atypical systemic amyloidosis in a patient who presented with recurrent subcutaneous bleeding of the eyelids and auricles. DESIGN: Observational case report. METHODS: A 52-year-old man with chronic cardiac failure, hypertension, weight loss, and impotence presented with recurrent bleeding of the eyelids and auricles, and was investigated for suspected amyloidosis. RESULTS: Biopsy specimens taken from the bone marrow, salivary gland, abdominal subcutaneous fat, and gastric mucosa, and eyelids were all negative for Congo red. Results of a conjunctival biopsy, which were returned after the patient's death, and cardiac muscle autopsy were positive for Congo red staining, indicating amyloidosis. CONCLUSION: The findings in this case show that the presentation of amyloidosis may be atypical. Ophthalmologists should consider a diagnosis of amyloidosis in patients with periorbital bleeding, even if biopsies are negative for Congo red. A conjunctival biopsy may be useful in establishing a diagnosis of primary systemic amyloidosis. PMID- 15364244 TI - Embolic cilioretinal artery occlusion due to carotid artery dissection. AB - PURPOSE: To report a case of embolic cilioretinal artery occlusion caused by carotid artery dissection. DESIGN: Interventional case report. METHODS: A 38-year old woman presented with acute visual loss in her right eye. Funduscopy showed a cilioretinal artery occlusion, which was confirmed by a fluorescein angiography. An embolus was found in the distal segment of the vessel. RESULTS: Color Doppler images of right internal carotid artery (ICA) disclosed a pseudolumen, suggesting a diagnosis of carotid dissection. Retrobulbar color Doppler image showed relative low flow velocity in the ophthalmic artery without flow reversal. Magnetic resonance angiography and cerebral angiogram showed total occlusion of the right ICA. Follow-up visual field examination revealed an inferior central defect fed by the cilioretinal artery. CONCLUSION: The pathogenesis of retinal artery occlusion caused by carotid dissection may be embolic or hemodynamic. In our case, a permanent visual defect was related to embolic occlusion of the cilioretinal artery. PMID- 15364245 TI - Primary Nocardia brasiliensis of the eyelid. AB - PURPOSE: To report a rare case of lymphocutaneous Nocardia brasiliensis originating in the eyelid. DESIGN: Observational case report. METHODS: The clinical presentation, workup, and treatment of a case of lymphocutaneous Nocardia brasiliensis originating in the eyelid are presented. RESULTS: The patient presented with a preseptal cellulitis from an abrasion of the eyelid that progressed to submandibular lymph node suppuration. Culture was performed, and a diagnosis of lymphocutaneous Nocardia brasiliensis was made. CONCLUSION: Nocardia brasiliensis may cause a lymphocutaneous infection of the face and must be considered in the differential diagnosis of preseptal cellulitis. PMID- 15364246 TI - Congenital eyelid imbrication syndrome. AB - PURPOSE: To describe congenital eyelid imbrication syndrome and its possible pathophysiology. DESIGN: Clinical observational case report. METHODS: A full-term newborn was examined after a vaginal delivery and uneventful pregnancy. RESULTS: The upper eyelids were overlapping the lower eyelids when the eyes were closed or when the newborn was asleep. The upper eyelids resumed normal position gradually over a week without causing any symptoms or residual sequelae. CONCLUSIONS: Eyelid imbrication syndrome is a rare cause of congenital eyelid malposition. It may be caused by inborn laxity of the upper medial and lateral canthal tendons that are tightened during the postnatal period, causing resolution of this condition. PMID- 15364247 TI - A simple device for teaching direct ophthalmoscopy to primary care practitioners. AB - PURPOSE: Ophthalmoscopy, a valuable skill for primary care practitioners, can be challenging to learn. A simple and inexpensive device for teaching direct ophthalmoscopy to primary care practitioners is described. DESIGN: Device description. METHODS: Cylindrical plastic canisters were altered to have an artificial pupil at one end and a replaceable fundus photograph at the other end to simulate the mechanics of performing direct ophthalmoscopy on a real eye. These were tested for ease of use by primary care students. RESULTS: The devices to aid in teaching ophthalmoscopy proved to be simple and inexpensive to construct. They allowed students to practice direct ophthalmoscopy technique and identification of funduscopic abnormalities. CONCLUSION: This simple device for teaching direct ophthalmoscopy to primary care practitioners is inexpensive to create and is a valuable aid for teaching direct ophthalmoscopy to primary care practitioners. PMID- 15364248 TI - Increased ocular pulse amplitude revealing aortic regurgitation. AB - PURPOSE: To report a case in which observation of increased ocular pulse amplitude (OPA) estimates in both eyes lead to a diagnosis of, and was presumably due to, aortic regurgitation. DESIGN: Case report. METHODS: An incidental finding of high OPA estimates in both eyes lead to a finding of widened arterial pulse pressure and aortic regurgitation. OPA estimates were taken after aortic valve surgery when arterial pulse pressure was normal. RESULTS: OPA estimates were 9 mm Hg in both eyes when arterial pulse pressure was high and 3 mm Hg in each eye when normal. CONCLUSION: Widened arterial pulse pressure may lead to increased OPA. PMID- 15364249 TI - A presumed missense mutation of RPGR causes abnormal RNA splicing with exon skipping. AB - PURPOSE: A patient with retinitis pigmentosa demonstrated a novel RPGR mutation (213G>A, last base of exon 2) predicted to cause a missense change (G52R) in the final protein. This study was performed to determine whether this mutation altered the effectiveness of the adjacent splice site. DESIGN: Observational case report. METHODS: Total RNA was extracted from leukocytes of the proband and his carrier mother. Reverse transcription-polymerase chain reaction (RT-PCR) was performed by using the primers flanking exon 2 of RPGR transcript, followed by gel purification and direct sequencing. RESULTS: Sequencing revealed skipping of exon 2 in the mutated transcript, leading to in-frame deletion of 42 amino acids affecting the critical RCC1-like domain. CONCLUSIONS: The last base of exons is conserved as "G" in 80% of splicing consensus sequences, yet when changed, can completely disrupt constitutive splicing as in this patient. Our data confirm that the evaluation of the effects of some DNA sequence alterations at the RNA level might have important implications for appropriate genotype-phenotype correlations. PMID- 15364250 TI - Deep anterior lamellar keratoplasty in a patient with descemetocele following gonococcal keratitis. AB - PURPOSE: To describe the presentation and subsequent management of a case of severe gonococcal keratitis in a young man. DESIGN: Case report. METHODS: A young man presented with severe gonococcal keratitis. Topical and systemic antibiotics were given with no improvement, and progressive corneal melting was noted. Deep lamellar keratoplasty was performed. The clarity and state of corneal graft and the postoperative visual acuity were noted. RESULTS: At 6 months posttreatment, the graft was clear, and no recurrence of infection was noticed. Corrected visual acuity was 20/25. CONCLUSION: Deep lamellar keratoplasty should be considered as a therapeutic option in patients with severe gonococcal keratitis that does not respond to antibiotic treatment. PMID- 15364251 TI - Thygeson's superficial punctate keratitis recurrence after laser in situ keratomileusis. AB - PURPOSE: To report a case of recurrent Thygeson's superficial punctate keratitis (SPK) in a patient submitted to photorefractive keratectomy (PRK) in one eye and laser in situ keratomileusis (LASIK) in the fellow eye. DESIGN: Observational case report. METHODS: A 32-year-old woman was diagnosed with Thygeson's SPK 5 years before undergoing PRK on the right eye and LASIK on the left eye to correct -3.00 + 1.50 x 20 and -3.75 sph, respectively. RESULTS: The patient presented with recurrence of Thygeson's SPK in the left eye 10 months after the LASIK procedure and no recurrence on the right eye, which was treated with PRK. CONCLUSIONS: This case report suggests that PRK may be a better option than LASIK in patients with ocular history of Thygeson's SPK. However, no strong conclusion can be established until further studies confirm the hypothesis of a lower recurrence rate of Thygeson's SPK following PRK compared with LASIK. PMID- 15364252 TI - Langerhans cell histiocytosis presenting as a limbal nodule in an adult patient. AB - PURPOSE: To report a case of Langerhans cell histiocytosis presenting as a limbal nodule. DESIGN: Observational case report. METHOD: A 26-year-old man presented with a superior limbal mass. On clinical examination, an infiltrative/neoplastic lesion was suspected, and a differential diagnosis of limbal amyloidosis, limbal fibrous histiocytoma, lymphoma, and juvenile xanthogranuloma was made. There was no other associated ocular or systemic abnormality. An excision biopsy was performed. RESULTS: The histopathology and immunohistochemical staining examination established the diagnosis of limbal Langerhans cell histiocytosis. CONCLUSION: To the best of our knowledge, this is an unusual presentation of Langerhans cell histiocytosis, and although rare, Langerhans cell histiocytosis should be included in the differential diagnosis of a limbal mass. PMID- 15364253 TI - Flattening of the posterior sclera: hypotony or elevated intracranial pressure? PMID- 15364255 TI - Comparison of the amblyopia treatment study HOTV and electronic-early treatment of diabetic retinopathy study visual acuity protocols in children aged 5 to 12 years. PMID- 15364257 TI - Photodynamic therapy for age-related macular degeneration. PMID- 15364259 TI - The monocular vertical prism dissociation test. PMID- 15364261 TI - Standardisation and controls, why can't we overcome the hurdles? PMID- 15364262 TI - Antiviral treatment and prophylaxis of influenza in primary care: German recommendations. AB - Antiviral drugs are a valuable supplementation to vaccines for the control and prevention of influenza. In Germany, for treating influenza amantadine, oseltamivir and zanamivir are approved. Amantadine and oseltamivir are also licensed for prophylactic use. On behalf of the Paul-Ehrlich-Society of Germany and the German Association for the Control of Virus Diseases, as two independent scientific societies, the first consensus Conference on the Antiviral Treatment and Prophylaxis of Influenza was held in June 2002. Based on the available data of clinical studies an expert group developed the following recommendations for the appropriate clinical use of the antiviral drugs: (1) since oseltamivir (orally administered) and zanamivir (administered by inhalation) have apparently similar clinical efficacy both drugs can be used alternatively for treatment. (2) Amantadine is not an alternative to the neuraminidase (NA) inhibitors because it is not effective against influenza B viruses, it frequently selects resistant virus mutants and it can cause adverse events. (3) When influenza is prevalent in the community patients with the clinical diagnosis of influenza should be treated with neuraminidase inhibitors if the symptoms are lasting not longer than 48 h. (4) Immunocompetent patients with a non-febrile illness and patients with a symptom history of more than 2 days should not be treated with antiviral drugs. (5) Although there are no data from clinical trials immunocompromised patients should also be treated when influenza has been diagnosed. (6) The prophylactic use of antiviral drugs can be recommended for persons with close contact to acutely ill persons and no recent vaccination against influenza. (7) The use of anti-influenza drugs have to be considered for prophylaxis in pandemics. A precondition for the adequate use of anti-influenza drugs in the primary medical care is the timely information on the local influenza situation delivered by surveillance systems. PMID- 15364263 TI - Application of quality systems to virology testing. PMID- 15364264 TI - Influenza A pneumonia presenting as progressive focal infiltrates in a stem cell transplant recipient. AB - BACKGROUND: Stem cell transplant recipients are susceptible to pulmonary infections, including influenza A. Typically, isolated influenza pneumonia has a diffuse, interstitial infiltrate pattern. OBJECTIVES: To describe the unusual clinical and radiographic course of influenza A pneumonia in a stem cell transplant recipient. STUDY DESIGN: Case report in which microbiologic assays, bronchoscopic and pathologic specimens are obtained. RESULTS: We describe a patient with influenza A pneumonia 8 months following a peripheral blood stem cell transplant who presented with minimal respiratory symptoms and rapidly progressing, focal pulmonary infiltrates. The large size and appearance of the masses have not been reported before in a patient with isolated influenza. CONCLUSION: This case highlights the differences of presentation and importance of early diagnosis and treatment of immunocompromised patients infected with influenza. PMID- 15364265 TI - Transcriptional analysis of human peripheral blood mononuclear cells after influenza immunization. AB - Influenza A virus is a major cause of morbidity and mortality worldwide. There is a large knowledge base on the immune response to influenza. However, few studies have focused on global gene expression in immune cells after antigenic challenge. A better understanding of the host immune response is required for the development of more efficient means of prevention and treatment of influenza. In this study, global gene expression in peripheral blood mononuclear cells (PBMCs) after influenza immunization was analyzed. The differential gene expression in antigen-stimulated and non-stimulated PBMCs was determined by cDNA microarrays. To determine whether a specific gene profile was present during a proliferative memory cell response to influenza antigens, gene expression in response to PHA was compared with antigen-stimulated PBMCs. PHA induced the upregulation of 201 genes while influenza virus antigen upregulated more than triple that is 630 genes out of 1700 genes analyzed. Both influenza antigen and PHA commonly upregulated 138 genes. Interferon (IFN)-related genes were induced by influenza but not by PHA. The interferon-gamma induced protein precursor 10 (IP-10) was upregulated 27-fold while the interferon-induced 54 kDa protein exhibited a 13 fold increase. The following gene families were also selectively upregulated by influenza antigens: complement ligands and receptors, T cell activation genes, growth factors, genes related to antigen processing and inflammatory responses. With PHA, the genes TNF-R, CTSG, CD3 delta, C8B, CRF1 and CCR2 had higher expression compared with the viral antigen stimulation. Neutrophil defensins alpha-1 and two C-C chemokines, proteins MIP-1-beta and MIP-4, were among the genes upregulated by both PHA and influenza antigens. The results suggest that interferon-induced genes are one of the main transcriptional targets during the immune response to influenza virus. PMID- 15364266 TI - Comparison of Binax NOW and Directigen for rapid detection of influenza A and B. AB - Directigen Flu A + B and Binax NOW Flu A and Flu B tests detected 33 (55.9%) and 31 (52.5%) of 59 influenza-positive samples, respectively. In children under 2 years of age, sensitivity increased to 75% for both tests. Three samples tested falsely-positive for influenza B using Binax NOW. PMID- 15364267 TI - Development of a PCR-and hybridization-based assay (PCR Adenovirus Consensus) for the detection and the species identification of adenoviruses in respiratory specimens. AB - BACKGROUND: Antigen detection assays and viral isolation techniques are routinely used to detect adenoviruses (Ad) associated with respiratory infections, and the value of the polymerase chain reaction (PCR) has recently been assessed. OBJECTIVES: This paper describes a PCR-hybridization-immunoenzymatic assay (PCR Adenovirus consensus) used to detect Ad and identify Ad species in respiratory specimens. RESULTS: On seven representative serotypes Ad 12, Ad 3, Ad 7, Ad 11, Ad 1, Ad 8, Ad 4, the mean genome equivalents per ml and the mean 50% infectious doses per ml were 10(6.3)and 10(4), respectively. Using 362 nasal aspirates from children, Ad were detected by immunofluorescence (IF) and culture in 97 cases (27%), by the PCR-Ad hexon method in 107 cases (29.5%) and by the PCR Adenovirus Consensus method in 113 cases (31.2%); 13 samples were found positive by both PCR and negative by the IF and culture methods; five samples were only positive according to the PCR Adenovirus Consensus) method. The sensitivity, specificity, predictive positive value and predictive negative value of the PCR Adenovirus Consensus method were 97.9%, 93.2%, 84%, 99.1%, respectively. The method identified the species (sp) from 91 positive amplicons: 1 Ad sp A, 44 Ad sp B, 42 Ad sp C, 3 Ad sp E, and 1 Ad sp F; 85 isolates were identified by IF or the neutralisation in culture, and 86 by a PCR-RE digestion method. The PCR Adenovirus Consensus detected six positive samples that were negative according to the IF and culture methods, and it identified the precise species of nine IF positive and culture-negative nasal aspirates. CONCLUSION: The PCR Adenovirus Consensus technique is more efficient than the classical IF or culture techniques for the detection of Ad in respiratory samples. An internal control is included to validate the screening results, and specific probes are used to identify the Ad species. PMID- 15364268 TI - Evaluation of quantitative and type-specific real-time RT-PCR assays for detection of respiratory syncytial virus in respiratory specimens from children. AB - BACKGROUND: Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract morbidity in young children and immunosuppressed patients. OBJECTIVES: To rapidly and accurately quantify and subtype RSV in respiratory samples, we developed and evaluated two real-time RT-PCR assays. STUDY DESIGN: A quantitative assay was designed using primers for a consensus region of the matrix protein gene and a subtype-specific assay for RSV-A and RSV-B detection was designed using primers for the polymerase gene. Quantitative RSV RT-PCR results of pediatric nasal wash samples submitted to the University of Washington Virology Laboratory from December 2002, through May 2003, were compared to those of an indirect fluorescent antibody RSV antigen detection assay (FA). RESULTS: Specificity of the RT-PCR assay was high, with no amplification of eleven common respiratory viruses and eight herpes viruses. Among 751 samples, RSV was detected in 267 (35.6%) by FA and in 286 (38.1%) by RT-PCR. Median RSV copy number in nasal wash samples that were positive by both FA and RT-PCR was 2.5 x 10(7) copies/mL versus a median of 3.0 x 10(4) copies/mL for samples positive by RT-PCR only (P < 0.001). The detection and quantity of RSV in respiratory specimens was associated with younger age, but not with gender or hospitalization. Among positive samples from this Seattle cohort, 52% were subtype A and 48% were subtype B. Both subtypes were detected with similar viral loads among all patient groups (stratified by age, gender, and hospitalization), and throughout the specimen collection period. CONCLUSIONS: These real-time RT-PCR assays provide a rapid, specific, and highly sensitive alternative for detecting, quantifying, and subtyping RSV in clinical specimens. PMID- 15364269 TI - Comparison of two new rapid antigen detection assays for respiratory syncytial virus with another assay and shell vial culture. AB - BACKGROUND: Respiratory syncytial virus (RSV) is the major viral pathogen causing lower respiratory tract infection in young children. Early detection allows cohorting of infected inpatients to prevent nosocomial transmission and consideration of treatment. To achieve rapid reporting, antigen detection has been widely used. Since late 2002, the FDA approved two new RSV antigen detection tests, the Now RSV test (Binax) and the Directigen EZ RSV (Becton Dickinson), both of which promised reduced hands-on processing. OBJECTIVES: Evaluate relative performance of the two new rapid antigen detection assays. STUDY DESIGN: To compare the performance of these newer tests with a previously existing antigen detection test (Becton Dickinson) and shell vial viral culture with clinical specimens. RESULTS AND CONCLUSIONS: A total of 89 fresh respiratory specimens from 84 pediatric patients were tested. We also performed reverse transcription polymerase chain reaction (RT-PCR) on all culture negative but antigen positive samples as well as 26 specimens that were negative on both shell vial culture and antigen testings. Overall, the three antigen assays performed similarly well. The hands-on processing for the two newer tests was significantly reduced, making them more convenient for rapid screening. PMID- 15364270 TI - Influenza circulating strains in Argentina exhibit differential induction of cytotoxicity and caspase-3 in vitro. AB - BACKGROUND: Human influenza infections are a significant cause of morbidity worldwide. Though damage to the respiratory epithelium and has been related to apoptosis, which occurs subsequent to influenza virus infection, little information is available regarding cell cytotoxicity of human strains. OBJECTIVE: To study cytotoxicity performed in vitro by various circulating strains in Argentina. The study sample consisted of three vaccine strains (H1N1, H3N2, and B) administered during 1999-2000 in South America and three strains isolated from clinical samples, one, NAC (H1N1) obtained from an adult inpatient with human pneumonia; and the other two (T) and (T2) (H3N2) with influenza syndrome. Viral antigen was detected by an immunofluorescence test, conducted prior to viral isolation in MDCK cells. Strains were subtyped by the hemmaglutination inhibition test. Cytotoxic properties were determined by lactate dehydrogenase reaction (LDH), crystal violet staining and Hoechst staining. Caspase-3 activity, morphological changes of apoptosis, and viral yields were measured in MDCK infected cells. RESULTS AND CONCLUSIONS: Cells infected by each of the strains exhibited apoptosis morphology by Hoechst staining and caspase-3 activity was high for both H1N1 strains. Further, high levels of LDH activity were detected for NAC and H3N2 strains tested, indicating the possible role of different viral proteins or functions on cell cytotoxicity. The NAC strain, isolated from human pneumonia and antigenically related to A/New Caledonia /20/99 (H1N1), was the highest cytotoxic strain and an excellent inducer of caspase-3 activity. In turn, no parameter was related to different viral yields. We conclude that human strains studied in this paper may be useful tools in the characterization of molecular determinants involved in viral cytopathogenicity. PMID- 15364271 TI - High risk HPV load estimated by Hybrid Capture II correlates with HPV16 load measured by real-time PCR in cervical smears of HPV16-infected women. AB - BACKGROUND: High risk human papillomavirus (HR-HPV) load determined by quantitative methods has already been considered as highly predictive of future development of high grade cervical lesions. Some studies also demonstrated that Hybrid Capture II (HCII) results can be considered as a reflection of HPV DNA load, while others did not. HCI assay, well suited for routine HR-HPV screening, is not especially dedicated for quantitative use. However, we have recently shown that women with high viral loads assessed by HCII were at increased risk of cervical precancer. OBJECTIVES: The aim of the study was to determine if the values given by the HCII assay can be considered as quantitative. STUDY DESIGN: We used a real-time PCR allowing precise quantification of both HPV16 genome and albumin gene to normalize the measuring HPV16 load in cervical cells and to compare the data with those obtained by HCIIin a series of 40 HR-HPV positive samples. RESULTS: Reproducibility of the HPV16 real-time PCR, assessed from nine independent experiments of serial dilutions of SiHa cell DNA, was reflected in coefficients of variation for standard curves of crossing point (Cp) values below 5%. The HPV16 loads with a broad individual variability were significantly related to the cumulative load estimated by HCII and did not depend on the cellularity of samples. CONCLUSIONS: We assume that the HCII values can be used as a quantitative measure of HR-HPV DNA, so long as cervical specimens are collected using standardized protocols. PMID- 15364272 TI - Detection of hepatitis C virus by a user-developed reverse transcriptase-PCR and use of amplification products for subsequent genotyping. AB - BACKGROUND: Detection, quantitation and genotyping of hepatitis C virus (HCV) are important in selecting appropriate therapy. Current commercially available HCV genotyping kits, including sequencing-based TRUGENE HCV 5'NC and hybridization based INNO-LiPA HCV II assays, rely on amplification products (amplicons) generated by HCV reverse-transcriptase (RT)-PCR methods such as the Roche AMPLICOR HCV test. METHODS: We developed a one-step RT-PCR assay to amplify and detect HCV RNA, and the resulting amplicons were used for HCV genotyping (TRUGENE). A total of 142 clinical samples were used to compare results from the RT-PCR/TRUGENE assay and those generated by the COBAS AMPLICOR and INNO-LiPA tests. RESULTS: Eighty-seven of 108 plasma specimens which were positive by AMPLICOR were also positive by the user-developed RT-PCR, giving a sensitivity of 100.0%. The RT-PCR detected 2 of 21 AMPLICOR-negative specimens and none of 34 HCV-EIA-negative serum specimens, giving a specificity of 96.4%. The 87 amplicons from the RT-PCR yielded HCV genotypes. HCV genotype results from both TRUGENE and INNO-LiPA were all in agreement. The TRUGENE and INNO-LiPA assays identified 69 (79.3%) and 47 (54.0%) specimens, respectively at the subtype level. HCV subtype information agreed by both assays in 34 of 36 (99.4%) specimens. One specimen with HCV genotypes 2 and 4 by INNO-LiPA was classified as a single genotype 2 by TRUGENE. CONCLUSION: Our data showed that the user-developed RT-PCR has comparable sensitivity and specificity for the detection of HCV in clinical specimens. The amplicons generated by the RT-PCR can be used for HCV genotyping by the sequencing-based TRUGENE assay. PMID- 15364273 TI - Japanese encephalitis virus (JEV) is an important cause of encephalitis among children in Cuddalore district, Tamil Nadu, India. AB - BACKGROUND: Japanese encephalitis (JE) is endemic in Cuddalore district, Tamil Nadu (TN), Southern India. The reports of JE cases from the local hospitals did not reflect the actual disease burden. It is likely that these cases were attending the nearby referral hospitals, for want of better treatment facilities. OBJECTIVES: Between July 2002 and February 2003, a pilot study was undertaken to examine whether JE was a component of paediatric acute encephalitis syndrome (AES) reported to two major referral hospitals adjacent to Cuddalore, and to map the distribution of the JE cases. STUDY DESIGN: A total of 58 hospitalized children [0-15 years] with AES were investigated. Other than the routine laboratory investigations, either CSF or sera or both [depending on the availability] collected from these children were analyzed at Center for Research in Medical Entomology, Madurai (TN) for JEV-antigen, antibody detection, virus isolation and virus genome detection by indirect immunofluorescence, MAC enzyme linked immunosorbent assay (ELISA), insect bioassay and by reverse transcriptase polymerase chain reaction (RT-PCR), respectively. RESULTS: JE was established in 17 (29%) of 58 AES cases; half of the AES cases [31/58, 53%] and 59% [10/17] of JE cases were confined to JE-endemic areas in Cuddalore district. The JE confirmation scored by different assays varied according to the clinical phase of the illness. The attack rate was high among the children aged 3-8 years. The monthly distribution of acute encephalitic syndrome cases followed the distribution of JE cases [coinciding with the rainy season in this region] suggesting encephalitis of JE origin. CONCLUSION: In JE-endemic areas, the actual JE burden can be estimated by the collection of JE case reports from the local hospitals and from the referral hospitals. Building of diagnostic facilities in hospitals for JE is necessary to achieve this goal. PMID- 15364274 TI - Acute infectious mononucleosis and coincidental measles virus infection. AB - BACKGROUND: Both Epstein-Barr and measles viruses (MV) cause immune suppression, and the association of the two viruses is evaluated as life threatening. The cell immune impairment caused by simultaneous Epstein-Barr and measles viral infections was responsible for the complicated course of the disease in all described previously reports and for unfavorable outcomes in most of the cases. Timely diagnosis of coincidental viral infections could be a useful predictor for the clinical course and complications. Diagnosis must be based on an accurate assessment of clinical, hematologic, serologic manifestations and supported by appropriate laboratory methods. Recognizing the infectious etiology of concomitant infections is important for both clinicians and epidemiologists. OBJECTIVE: To describe a case report of a 20-year-old woman previously vaccinated against measles infected with acute mononucleosis and coincidental measles virus infection. STUDY DESIGN: The clinical, routine laboratory, as well as serological and virologic findings of this patient were scrutinized. Special emphasis was placed on the use of RT-PCR/PCR for confirming the involvement of both measles virus and Epstein-Barr virus (EBV) in this patient's illness. RESULTS: Infectious mononucleosis was not suspected at admission to the hospital. The final diagnosis of a concomitant measles virus infection and acute infectious mononucleosis was facilitated using viral serology to detect virus-specific IgG and IgM antibodies and by RT-PCR for the detection of measles virus RNA and EBV DNA from peripheral blood monocyte cells (PBMC). CONCLUSION: The present report highlights the difficulty of diagnosing two coincidental virus infections on clinical grounds. Serological and molecular laboratory methods, specifically the PCR (RT-PCR) analysis, are found to be useful for confirming the concomitant viral infections and proper identification of the infecting pathogens. PMID- 15364275 TI - The effect of some tannins on trout erythrocytes exposed to oxidative stress. AB - In order to gain more knowledge on the role of tannins as antioxidants, their ability to protect (Salmo irideus) erythrocytes against oxidative stress was investigated. Antioxidant activity of different tannins (tannic, gallic and ellagic acid) was evaluated by chemiluminescence (CL) techniques using lucigenin and luminol as chemiluminogenic probes for the superoxide radical generated by the xanthine/xanthine oxidase system and hydrogen peroxide, respectively. The superoxide-scavenging activity of these tannins was shown for all the compounds; however, it is not clear if this is due to their ability of scavenging the superoxide radical or to their inhibitory activity on xanthine oxidase. Tannic and ellagic acid showed a marked effect on the reduction of H2O2-luminol chemiluminescence. The influence of these tannins on the rate of hemolysis in stressed trout erythrocytes was investigated and the results indicate that tannic acid accelerates the hemolytic event while gallic and ellagic acid have no significant effect. The possible protective action of these compounds against oxidative DNA damage was assessed using the comet assay, a rapid and sensitive single-cell gel electrophoresis technique, used to detect primary DNA damage in individual cells. The results here reported show that tannins under study are capable at low concentrations of protecting DNA breakage, while at high concentrations they can be genotoxic. PMID- 15364276 TI - Disturbed microtubule function and induction of micronuclei by chelate complexes of mercury(II). AB - Interactions of mercury(II) with the microtubule network of cells may lead to genotoxicity. Complexation of mercury(II) with EDTA is currently being discussed for its employment in detoxification processes of polluted sites. This prompted us to re-evaluate the effects of such complexing agents on certain aspects of mercury toxicity, by examining the influences of mercury(II) complexes on tubulin assembly and kinesin-driven motility of microtubules. The genotoxic effects were studied using the micronucleus assay in V79 Chinese hamster fibroblasts. Mercury(II) complexes with EDTA and related chelators interfered dose-dependently with tubulin assembly and microtubule motility in vitro. The no-effect concentration for assembly inhibition was 1 microM of complexed Hg(II), and for inhibition of motility it was 0.05 microM, respectively. These findings are supported on the genotoxicity level by the results of the micronucleus assay, with micronuclei being induced dose-dependently starting at concentrations of about 0.05 microM of complexed Hg(II). Generally, the no-effect-concentrations for complexed mercury(II) found in the cell-free systems and in cellular assays (including the micronucleus test) were identical with or similar to results for mercury tested in the absence of chelators. This indicates that mercury(II) has a much higher affinity to sulfhydryls of cytoskeletal proteins than to this type of complexing agents. Therefore, the suitability of EDTA and related compounds for remediation of environmental mercury contamination or for other detoxification purposes involving mercury has to be questioned. PMID- 15364277 TI - Genotoxicity of diphenyl diselenide in bacteria and yeast. AB - Diphenyl diselenide (DPDS) is an electrophilic reagent used in the synthesis of a variety of pharmacologically active organic selenium compounds. This may increase the risk of human exposure to the chemical at the workplace. We have determined its mutagenic potential in the Salmonella/microsome assay and used the yeast Saccharomyces cerevisiae to assay for putative genotoxicity, recombinogenicity and to determine whether DNA damage produced by DPDS is repairable. Only in exponentially growing cultures was DPDS able to induce frameshift mutations in S. typhimurium and haploid yeast and to increase crossing over and gene conversion frequencies in diploid strains of S. cerevisiae. Thus, DPDS presents a behavior similar to that of an intercalating agent. Mutants defective in excision resynthesis repair (rad3, rad1), in error-prone repair (rad6) and in recombinational repair (rad52) showed higher than WT-sensitivity to DPDS. It appears that this compound is capable of inducing single and/or double strand breaks in DNA. An epistatic interaction was shown between rad3-e5 and rad52-1 mutant alleles, indicating that excision-resynthesis and strand-break repair may possess common steps in the repair of DNA damage induced by DPDS. DPDS was able to enhance the mutagenesis induced by oxidative mutagens in bacteria. N acetylcysteine, a glutathione biosynthesis precursor, prevented mutagenesis induced by DPDS in yeast. We have shown that DPDS is a weak mutagen which probably generates DNA strand breaks through both its intercalating action and pro-oxidant effect. PMID- 15364278 TI - Hematite (Fe2O3) acts by oxydative stress and potentiates benzo[a]pyrene genotoxicity. AB - Since epidemiological studies have implicated the co-exposition of iron oxides and polycyclic aromatic hydrocarbons as potential etiological factors involved in the excess of mortality from lung cancer in miners, experimental studies have been performed to investigate the role of iron on benzo[a]pyrene (B[a]P)-induced lung pathogenesis. We demonstrated previously that in vivo damage was higher when B[a]P was coated onto hematite than when B[a]P was administered alone. In order to determine the role of (i) different cell types and (ii) adsorption of hematite in this potentiation, in vitro studies were developed. The Comet assay was first used to measure DNA damage in four isolated cell types from Sprague-Dawley rats at 1, 2, 4, 8 and 24h after in vitro treatment with hematite (Fe2O3) or B[a]P or B[a]P coated onto hematite. For the two treatments with B[a]P, no damage was observed in alveolar macrophages, but significant increases in damage were seen in lymphocytes, hepatocytes and lung cells (where the effects of B[a]P coated onto hematite were stronger than those of B[a]P alone). In a second part of the study, the Comet assay was conducted with lung cells to measure the in vitro effect of (i) the coating and (ii) the role of the physical properties of Fe2O3. A statistically significant increase in damage was observed for the coating of B[a]P onto Fe2O3 compared (i) with their simple addition and (ii) with the coating of B[a]P onto graphite used as an inert compound. This study showed that (i) Fe2O3/B[a]P acts essentially in lung cells, (ii) the coating is a primordial step and (iii) the physical properties of Fe2O3 play a very minor role, which suggests another mechanism of action to explain the higher toxicity. Hence, our data may contribute to explain the excess of mortality in epidemiological studies and overall why exposures to B[a]P coated onto Fe2O3 resulted in higher toxicity in rodents compared to exposure to B[a]P alone. PMID- 15364279 TI - Sensitivity to DNA-damage induction and chromosomal alterations in mucosa cells from patients with and without cancer of the oropharynx detected by a combination of Comet assay and fluorescence in situ hybridization. AB - In addition to exogenous risk factors, the development of head and neck cancer is based on genetic alterations and individual sensitivity to mutagens. The DNA damaging effect of xenobiotics and the location of chromosomal changes warrant further investigation. The aim of this study was to evaluate variance in structural genetic changes in human epithelia as target cells for head and neck carcinogenesis. The combination of the single-cell gel electrophoresis (Comet) assay with the fluorescence in situ hybridization (FISH) technique is presented to examine differences in sensitivity to DNA-damage induction and in alterations of chromosomes 1, 3, 5 and 8 in patients with and without squamous cell carcinoma of the oropharynx. Macroscopically healthy biopsies from the mucosa, taken at a distance from the tumor of 10 patients with oropharyngeal carcinoma and from 10 patients without tumor were harvested during surgery. Cells were isolated by enzymatic digestion and incubated with benzo[a]pyrene-diolepoxide (BPDE), causing DNA-adduct formation by covalent binding of BPDE with DNA bases. The cells were subsequently analyzed by means of the Comet assay to separate DNA fragments and to visualize the DNA-damage. A hybridization mixture with whole-chromosome paints for Chr1, Chr3, Chr5 and Chr8 was added. After fluorescent staining, the entire DNA and the DNA of chromosomes 1, 3, 5 and 8 were evaluated by digital analysis. BPDE caused significant DNA damage in oropharyngeal mucosa cells of patients with and patients without carcinoma. No differences in the amount of DNA damage could be observed between patients suffering from sqamous cell carcinoma and patients without malignancy. Evaluation of chromosomal alterations, however, revealed significantly higher damage levels in chromosomes 3, 5 and 8 compared with chromosome 1 in tumor patients. In contrast, for patients without oropharyngeal carcinoma no differences in chromosomal alterations could be observed. The Comet assay could be combined with FISH to examine the sensitivity to DNA-damage induction and chromosomal alterations in human epithelial cells exposed to a genotoxic agent. Chromosomal breakage is increased for chromosomes 3, 5 and 8 as compared with chromosome 1, indicating a higher sensitivity of these chromosomes in epithelial cells of tumor patients. Using Comet/FISH on human epithelia, selected genetic alterations can be detected, which supports description of endogenous risk factors in carcinogenesis of the upper aerodigestive tract. PMID- 15364280 TI - Induction of apoptosis and G2/M arrest by N-methyl-N'-nitro-N-nitrosoguanidine in human prostate carcinoma cells. AB - We have investigated the effects of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), a well known DNA alkylating agent, on the growth and cell cycle progression in human prostate carcinoma PC-3 and DU145 cells, which are lacking both p53 alleles and having mutated p53, respectively. It was found that MNNG could inhibit the cell growth in a dose-dependent manner, which was associated with dendrite-like morphological change and induction of apoptotic cell death. Flow cytometry showed that MNNG could cause an arrest at the G2/M phase of the cell cycle, which is closely correlated to inhibition of cyclin-dependent kinase (Cdk) 2 and Cdc2 kinase activities. Furthermore, this compound induced Cdk inhibitor p21WAF1/CIP1 expression at both the transcription and protein levels in a p53-independent manner. MNNG also activated the reporter construct of a p21 promoter. Present results indicate that the up-regulation of p21 by MNNG is likely responsible for the inhibition of Cdks kinase activity rather than the down-regulation of cyclins and Cdks expression. PMID- 15364281 TI - Genotoxicity evaluation of individuals working with photocopying machines. AB - Photocopying machines are a common sight in the cities of India. There is ample evidence showing that the components of toners individually or in the form of a complex mixture are genotoxic. Toxic components of the photocopiers are from their emissions, toners and extremely low frequency electromagnetic fields (ELF EMFs). In the present study micronucleus test (MNT) on buccal epithelial cells, cytokinesis block micronucleus (CBMN) assay and chromosomal aberration analysis on peripheral blood mononuclear cells was performed on 98 workers occupationally involved in photocopying and 90 age and sex matched controls. The results showed a significant increase in the frequency of MN in buccal epithelial cells and peripheral blood lymphocytes, as well as chromosomal aberrations in the exposed as compared to the control subjects. PMID- 15364282 TI - Mutagen X and chlorinated tap water are recombinagenic in yeast. AB - This study determines the effects of a water disinfection by-product, 3-chloro-4 (dichloromethyl)-5-hydroxy-2(5H)-furanone (also known as mutagen X or MX) and chlorinated tap water on genomic instability in the yeast Saccharomyces cerevisiae. Tap water samples collected from Cherepovets (Russia) and Boston (MA, USA), were extracted using XAD absorption and ethyl acetate elution. MX and these water extracts were then tested for their ability to induce intrachromosomal recombination (deletions or DEL events), interchromosomal recombination (ICR) and aneuploidy (ANEU) using the yeast DEL assay. MX strongly induced DEL, ICR and ANEU events with a positive dose response and no threshold. Cherepovets tap water induced DEL and ICR events while evidence of ANEU induction was weak. The DEL induction potencies were stronger at higher concentrations. The estimated contribution of MX to DEL induction varied from over 50% at low concentrations (which is comparable to a typical contribution of MX to Ames mutagenicity of tap water) to between 2 and 10% at highest concentrations. For Boston tap water, there was only weak evidence of DEL induction and no evidence of ICR and ANEU induction. This is consistent with the results of other studies, which reported much higher concentrations of MX and stronger Ames mutagenicity in Cherepovets tap water than in Boston tap water. PMID- 15364283 TI - Purification and characterization of alpha-amylase from the infective juveniles of the nematode Heterorhabditis bacteriophora. AB - Glycogen content and alpha-amylase activity were estimated in the infective juveniles (IJs) of Heterorhabditis bacteriophora at different times of storage. The glycogen content declined from 5.8 to 2.5 ng/IJ during storage for 40 days at 27 degrees C. The change in glycogen content coincided with the change of alpha amylase activity during storage. alpha-Amylase was purified from IJs at zero time of storage by ion exchange chromatography and gel filtration. Ion exchange chromatography resolved alpha-amylase into three isoenzymes. The major isoenzyme alpha-amylase I had the highest specific activity and was purified to homogeneity. A molecular mass of 46-47 kDa was estimated for both the native and denatured enzyme, suggesting that the enzyme is monomeric. The Km values were 6.5 and 9.6 mg/ml using starch and glycogen as substrates, respectively. alpha Amylase I showed optimum activity at pH 7.0 and had an optimum temperature of 40 degrees C. The enzyme was unstable at temperatures above 40 degrees C. The enzyme activity was severely inhibited by EDTA, p-CMB and iodoacetic acid, but potentiated by CaCl2 and NaCl. These results are discussed and compared with previously reported alpha-amylases in the insect hosts of the parasite. PMID- 15364284 TI - Purification and characterization of lysozyme from plasma of the eastern oyster (Crassostrea virginica). AB - Lysozyme was purified from the plasma of eastern oysters (Crassostrea virginica) using a combination of ion exchange and gel filtration chromatographies. The molecular mass of purified lysozyme was estimated at 18.4 kDa by SDS-PAGE, and its isoelectric point was greater than 10. Mass spectrometric analysis of the purified enzyme revealed a high-sequence homology with i-type lysozymes. No similarity was found however between the N-terminal sequence of oyster plasma lysozyme and N-terminal sequences of other i-type lysozymes, suggesting that the N-terminal sequences of the i-type lysozymes may vary to a greater extent between species than reported in earlier studies. The optimal ionic strength, pH, cation concentrations, sea salt concentrations, and temperature for activity of the purified lysozyme were determined, as well as its temperature and pH stability. Purified oyster plasma lysozyme inhibited the growth of Gram-positive bacteria (e.g., Lactococcus garvieae, Enterococcus sp.) and Gram-negative bacteria (e.g., Escherichia coli, Vibrio vulnificus). This is a first report of a lysozyme purified from an oyster species and from the plasma of a bivalve mollusc. PMID- 15364285 TI - Phospholipase A2 in salivary glands isolated from tobacco hornworms, Manduca sexta. AB - We describe a phospholipase A2 (PLA2) associated with the salivary glands of tobacco hornworms, Manduca sexta. This enzyme is able to hydrolyze arachidonic acid from the sn-2 position of PLs. Addition of the calcium chelator, EGTA, or calcium, to the Tris reaction buffer impaired the PLA2 activity, from which we infer the enzyme requires very low concentrations of calcium or perhaps other ions for optimal activity. PLA2 activity was sensitive to protein concentration (highest activity at 25 microg protein per microl), reaction time (optimal at 30 min), buffer pH (optimal at pH 8-10), and reaction temperature (optimal range 18 38 degrees C). The salivary gland PLA2 was sensitive to the site-specific inhibitor, oleyloxyethylphosphorylcholine and stable to freezing at -80 degrees C, but not -20 degrees C. The biological significance of this enzyme may relate to hydrolysis of fatty acid moieties from dietary PLs for absorption by midgut epithelia. This salivary gland enzyme may also be responsible for killing food borne bacteria. PMID- 15364286 TI - Molecular cloning and characterization of the translationally controlled tumor protein gene in Bombyx mori. AB - Translationally controlled tumor protein (Tctp/p23) is known to be synthesized preferentially in cells during the early growth phase of tumors, but is also expressed in normal cells. To elucidate its molecular basis of the expression and physiological significance, a cDNA encoding for the Bombyx mori Tctp (BmTctp) was deduced by editing the partial cDNA sequences registered in a Bombyx EST database. RT-PCR analyses indicated that the BmTCTP mRNA was transcribed in all larval organs examined and was present constantly during the cell cycle of BmN4 cells. A genomic clone of 4255 nucloetide residues produced by inverse PCR contained the 5'-flanking region, two introns and three exons of the BmTCTP gene. Sequence analysis of the 5'-flanking region indicated that a putative promoter region contains several canonical transcription elements such as GATA box, CCAAT motif, MEF2, E4BP4.01 and AP-1, but lacks a TATA box element. Luciferase reporter assay of the deletion constructs of the 5'-flanking region revealed that the -676 to +66 region enhanced the promoter activity the most markedly. In addition to this, there were at least two enhancer-like elements and several repressor elements. PMID- 15364287 TI - The evolution of carotenoid coloration in estrildid finches: a biochemical analysis. AB - The estrildid finches (Aves: Passeriformes: Estrildidae) of Africa, Asia, and Australia have been the focus of several recent tests of sexual selection theory. Many estrildids display bright red, orange, or yellow colors in the beak or plumage, which typically are generated by the presence of carotenoid pigments. In this study, we used high-performance liquid chromatography to investigate the carotenoid content of feathers and other colorful tissues in seven species of estrildids. Star finches (Neochmia ruficauda) and diamond firetails (Stagonopleura guttata) circulated two main dietary carotenoids (lutein and zeaxanthin) through the blood and liver and used both to acquire a yellow plumage color. However, five other estrildids (common waxbill, Estrilda astrild; black rumped waxbill, Estrilda troglodytes; zebra waxbill, Amandava subflava; red avadavat, Amandava amandava; and zebra finch, Taeniopygia guttata) circulated these same dietary carotenoids along with two metabolites (dehydrolutein and anhydrolutein) through the blood and/or liver and used all four as yellow plumage colorants. We subsequently tracked the distribution of these pigments using a published phylogeny of estrildid finches to determine the evolutionary pattern of carotenoid metabolism in these birds. We found that finches from the most ancient tribe of estrildids (Estrildini) possessed the ability to metabolize dietary carotenoids. Although carotenoids from the most ancestral extant estrildid species have yet to be analyzed, we hypothesize (based on their relationships with other songbirds known to have such metabolic capabilities) that these finches inherited from their ancestors the capability to metabolize carotenoids. Interestingly, later in estrildid evolution, certain taxa lost the ability to metabolize dietary carotenoids (e.g., in the Poephilini), suggesting that the occurrence of carotenoid metabolism can be labile and is likely shaped by the relative costs and benefits of color signaling across different species. PMID- 15364288 TI - Fatty acyl CoA synthetase from Antarctic notothenioid fishes may influence substrate specificity of fat oxidation. AB - Antarctic notothenioid fishes possess large lipid stores that are important fuels for aerobic metabolism. Oxidative muscle tissues of these animals oxidize long chain mono-unsaturated fatty acids more readily than saturated fatty acids. The mechanistic basis(es) for the substrate specificity of their fatty acid-oxidizing pathway is unknown. We examined the substrate specificity of fatty acyl coenzyme A synthetase (FACS) to determine whether the enzyme contributes to targeting unsaturated fatty acids for preferential transport into mitochondria as fuels for beta-oxidation. Maximal activities of FACS were measured in isolated mitochondria from Notothenia coriiceps and Chaenocephalus aceratus oxidative skeletal muscles in the presence of fatty acids differing in chain lengths and degrees of unsaturation. With the exception of C(22:6), maximal activities were greater with unsaturated substrates than with C(16:0), a saturated fatty acid. Monoenoic fatty acids did not produce the highest activities. Predicted amino acid sequences of FACS from Antarctic C. aceratus, Gobionotothen gibberifrons, and N. coriiceps and sub-Antarctic Notothenia angustata and Eleginops maclovinus were determined to identify amino acid candidates that may be important for determining the substrate specificity of FACS. Substitutions cysteine548 and polar threonine552 within the putative fatty acid binding pocket may contribute to preference for unsaturated fatty acyl substrates compared to saturated fatty acids. PMID- 15364289 TI - Isolation and molecular characterization of cathepsin L-like cysteine protease cDNAs from western flower thrips (Frankliniella occidentalis). AB - Cysteine proteases are predominant in thrips guts (TGs) and, therefore, a suitable target for selecting effective protease inhibitors against western flower thrips (Frankliniella occidentalis). We report the isolation of four full length cysteine protease cDNA clones from thrips in a two-step PCR approach with degenerate oligonucleotides designed on conserved cathepsin L domains. At the deduced amino acid level, the clones possessed all functional and structural characteristics of cathepsin L, and showed high mutual identity and strong similarity with cathepsin L-like cysteine proteases from other insects and arthropods. Southern analysis indicated that a family of four closely related and 10-12 less-related genes encode the cathepsin L-like cysteine proteases in the thrips genome. Partial sequencing of genomic DNA demonstrated the presence of three introns in the coding DNA. PMID- 15364290 TI - A novel enzymatic reaction for converting DNA to CO-DNA. AB - We have found previously that DNA from both the chick cerebrum and cardiac muscle has a modified structure. We named this novel DNA, CO-DNA. CO-DNA is a form of DNA in which a carbonyl group is attached to C-1 of the 2-deoxyribose and to the nitrogenous base. Therefore, 3-deoxyglucosone is the sugar constituent for CO DNA. We found previously that the modification of the sugar moiety in DNA occurs around embryonic day 12 in the chick embryo. In this study, we isolated enzymes for the conversion of DNA to CO-DNA from chick cerebra. In our reaction system, uniformly labeled 14C-glucose was used as substrate. During incubation, the radioactivity was incorporated into DNA. From the analysis of 14C-labeled deoxynucleoside, the radioactive sugar was confirmed to be 3-deoxyglucosone. We propose a series of reactions involved in the conversion of DNA to CO-DNA: (1) DNA-enzyme complex is formed during preincubation, (2) 14C-glucose is transformed to 14C-3-deoxyhexonic acid, (3) 14C-3-deoxyhexonic acid is subsequently transformed into the sugar-phosphate, which is a mixture of phosphorylated 14C-3 deoxyhexonic acid and phosphorylated 14C-3-deoxyglucosone, (4) 2-deoxyribose in DNA is replaced with 14C-3-deoxyglucosone through its intermediate phosphorylated form, and (5) DNA is finally converted to CO-DNA. PMID- 15364291 TI - In vitro effects of mammalian leptin, neuropeptide-Y, beta-endorphin and galanin on transcript levels of thyrotropin beta and common alpha subunit mRNAs in the pituitary of bighead carp (aristichthys nobilis). AB - Thyrotropin (thyroid stimulating hormone, TSH) is a member of the pituitary glycoprotein hormones, consisting of two dissimilar subunits, alpha and beta. The two subunits are produced by different genes and are regulated independently. We have previously cloned a TSHbeta cDNA from bighead carp pituitary and investigated its gene regulation. We report here the direct effects of mammalian TSH-releasing hormone (TRH), leptin, neuropeptide-Y (NPY), beta-endorphin and galanin on mRNA levels of both TSHbeta and alpha-subunits in the pituitary of bighead carp in vitro. The dispersed pituitary cells of bighead carp were incubated at 25 degrees C for 6 h with different doses of these factors. The relative mRNA levels of TSHbeta and alpha-subunits were estimated by traditional polymerase chain reaction (PCR) analysis and fluorescence real-time PCR analysis. The results revealed that mammalian TRH, leptin and beta-endorphin produced dose dependent stimulatory effects on mRNA levels of both TSHbeta and alpha-subunits while thyroxine (T4) and mammalian galanin suppressed mRNA levels of both TSHbeta and alpha-subunits. NPY suppressed TSHbeta mRNA level, but stimulated alpha subunit mRNA level. This study has demonstrated that mammalian TRH, leptin, NPY, beta-endorphin and galanin were active in modulating the steady-state mRNA levels of TSHbeta and alpha-subunits of bighead carp pituitary in vitro. The results suggest that endogenous TRH, leptin, NPY, beta-endorphin and galanin may modulate transcript levels of TSHbeta and alpha-subunits in pituitary of bighead carp. PMID- 15364292 TI - In vivo DNA double-strand breaks enhance gene targeting in cultured silkworm cells. AB - Alteration of genomic information through homologous recombination (HR) is a powerful tool for reverse genetics in bacteria, yeast, and mice. The low frequency of HR is, however, a major obstacle to achieve efficient gene targeting. In this study, we have developed an assay system for investigating the frequency of gene targeting in cultured silkworm cells using a firefly luciferase gene as a reporter. The introduction of a DNA double-strand break (DSB) either in the chromosomal target locus or in the targeting construct drastically increased the frequency of gene targeting. Interestingly, the inhibition of poly(ADP ribose) polymerase (PARP), a protein known to play an important role in overall suppression of the HR pathway, stimulated the targeting efficiency, whereas the overexpression of two silkworm RecA homologs, BmRad51 and BmDmc1, had no effect. The presently devised assay system may serve as a useful tool to improve the gene targeting efficiency in the silkworm (Bombyx mori). PMID- 15364293 TI - cDNA cloning, expression, and enzymatic activity of a cellulase from the mulberry longicorn beetle, Apriona germari. AB - A novel cellulase [beta-1,4-endoglucanase (EGase), EC 3.2.1.4] cDNA belonging to glycoside hydrolase family (GHF) 45 was cloned from the mulberry longicorn beetle, Apriona germari. The cDNA encoding EGase of A. germari (Ag-EGase) is 711 bp long with an open reading frame of 237 amino acid residues. The Ag-EGase was closely related to another beetle, Phaedon cochleariae, cellulase and one symbiotic protist cellulase in the hindgut of the termite Reticulitermes speratus, those belonging to GHF 45. The catalytic sites of GHF 45 are conserved in Ag-EGase. Southern blot analysis of genomic DNA suggested the presence of Ag EGase gene as a single copy and Northern blot analysis confirmed midgut-specific expression at transcriptional level. Similarly, the Ag-EGase enzyme assay exhibited high activity only in midgut tissue, suggesting that the midgut is the prime site where large quantities of EGase are synthesized for degrading the absorbed cellulose from the diet. The cDNA encoding Ag-EGase was expressed as a 29-kDa polypeptide in baculovirus-infected insect Sf9 cells and the culture supernatants of the recombinant baculovirus-infected cells showed EGase enzyme activity of 15.25 U/ml of medium containing 0.5 x 10(6) cells at 5 days post infection (p.i.). The enzyme activity of the purified recombinant Ag-EGase expressed in baculovirus-infected insect cells was approximately 992 U per mg of recombinant Ag-EGase. The purified recombinant Ag-EGase showed the highest enzymatic activity at 50 degrees C and pH 6.0, and was stable at 55 degrees C at least for 10 min. PMID- 15364294 TI - A novel disintegrin, jerdonatin, inhibits platelet aggregation and sperm-egg binding. AB - A novel disintegrin, jerdonatin, was purified to homogeneity from Trimeresurus jerdonii venom by gel filtration and reversed-phase high-pressure liquid chromatography. We isolated the cDNA encoding jerdonatin from the snake venom gland. Jerdonatin cDNA precursor encoded pre-peptide, metalloprotease and disintegrin domain. Jerdonatin is composed of 72 amino acid residues including 12 cysteines and the tripeptide sequence Arg-Gly-Asp (RGD), a well-known characteristic of the disintegrin family. Molecular mass of jerdonatin was determined to be 8011 Da by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Jerdonatin inhibited ADP- and collagen induced human platelet aggregation with IC50 of 123 and 135 nM, respectively. We also investigated the effect of jerdonatin on the binding of B6D2F1 hybrid mice spermatozoa to mice zona-free eggs and their subsequent fusion. Jerdonatin significantly inhibited sperm-egg binding in a concentration-dependent manner, but had no effect on the fusion of sperm-egg. These results indicate that integrins on the egg play a role in mammalian fertilization. PMID- 15364295 TI - Estrogen synthesis in relation to gonadal development of Japanese scallop, Patinopecten yessoensis: gonadal profile and immunolocalization of P450 aromatase and estrogen. AB - Aromatase activities and estrogen contents in the gonad of Japanese scallop, Patinopecten yessoensis, were determined during gonadal development and estrogenic cells in the testis were identified immunohistochemically. Ovaries and testes developed rapidly during January and February to reach the mature stage in March and the spawning stage in April. Increases in aromatase activities of the ovary and testis preceded the onset of the ovarian and testicular development. Aromatase activities reached the highest level at the growing stage in February and the mature stage in March, and showed a striking decrease at the spawning stage in April. Contents of ovarian and testicular estradiol-17beta changed similarly to the profile of aromatase activities in the ovary and testis, although estrone showed no change. Immunoreactivities against P450 aromatase and estradiol-17beta were detected in the cells along the inside of the acinar wall of the testis, whereas in the previous reports, the cells are distributed along the outside of the acinar wall in the ovary. This study thus suggests that estrogen is synthesized in the estrogenic cells of the ovary and testis through aromatization by P450 aromatase and that testicular estrogen may play a physiological role in spermatogenesis. PMID- 15364297 TI - Proceedings of the 8th International Symposium on Clinical Evaluation of Drug Efficacy in Urinary Tract Infection and the symposium "Urinary Tract Infections" organised by the Commission for Urinary Tract Infection of the International Society of Chemotherapy. June 7-9, 2003. Durban, South Africa. PMID- 15364296 TI - Carotenoids of two freshwater amphipod species (Gammarus pulex and G. roeseli) and their common acanthocephalan parasite Polymorphus minutus. AB - Carotenoid compositions of two freshwater Gammarus species (Crustacea: Amphipoda) and of their common acanthocephalan parasite Polymorphus minutus were characterized. The effect of carotenoid uptake by the parasite was addressed by comparing the carotenoid content of uninfected and infected female hosts. Using high-pressure liquid chromatography (HPLC), co-chromatography of reference pigments and electron ionization mass spectrometry of collected HPLC fractions (EI-MS), several xanthophylls and non-polar compounds were identified. Seven kinds of carotenoids, mainly xanthophylls, were identified in gammarids. Astaxanthin was predominant, amounting to 40 wt.% of total carotenoid in both uninfected G. pulex and G. roeseli. By contrast, we found only non-polar compounds with a predominance of esterified forms of astaxanthin in P. minutus larvae. No significant effect of infection on carotenoid content was evidenced in G. pulex and G. roeseli females. Our study highlights the use of a Matrix Solid Phase Dispersion as an efficient extraction method of both xanthophylls and non polar pigments in small samples, including lipid-rich ones as P. minutus parasite. We discuss on the presumptive pathway leading to the formation of free astaxanthin in gammarids via hydroxy compounds, and on the accumulation of esters of astaxanthin in parasites. PMID- 15364298 TI - Management of vaginal discharge syndrome: how effective is our strategy? AB - Although syndromic management of sexually transmitted diseases in highly endemic areas provides a short-term benefit to the individuals treated, it has no impact on decreasing prevalence rates. The numerous factors that contribute to this are discussed. Rapid reinfection from a large pool of infected symptomatic and asymptomatic individuals as well as the non-specific nature of presenting symptoms in women with vaginal discharge syndrome are major causes. PMID- 15364299 TI - Aetiology of male urethritis in patients recruited from a population with a high HIV prevalence. AB - The aetiology of urethritis, the significance of potential pathogens and the relation of urethritis to HIV infection were determined in 335 men (cases) with and 100 men (controls) without urethral symptoms. Urethral swab specimens were tested for different organisms by PCR or by culture for Neisseria gonorrhoeae. The prevalence of N. gonorrhoeae and Chlamydia trachomatis was 52 and 16%, respectively. The potential pathogens: Mycoplasma genitalium, Ureaplasma urealyticum, Trichomonas vaginalis and herpes simplex virus (HSV), were present in 5, 36, 6 and 6% of the cases respectively. M. genitalium was the only potential pathogen associated with microscopic urethritis. After excluding gonococcal infections, U. urealyticum was more frequent in symptomatic patients, while the prevalence of T. vaginalis was similar among cases and controls. These results strongly suggest an a etiological role for M. genitalium in male urethritis, a possible role for U. urealyticum, but not for T. vaginalis. The control group, with 97% genital ulcer disease patients, was not suitable for the investigation of the role of HSV. The sero-prevalence of HIV was 45%. Current infections were not associated with HIV. However, a history of previous urethral discharge was associated with HIV in a multivariate analysis and supported the hypothesis that non-ulcerative sexually transmitted diseases facilitate HIV transmission. PMID- 15364300 TI - Antimicrobial resistance of Neisseria gonorrhoeae in Japan, 1993-2002: continuous increasing of ciprofloxacin-resistant isolates. AB - Susceptibility testing was conducted on 1357 isolates of Neisseria gonorrhoeae isolated from 1993 through 2002 in Japan to assess the antimicrobial resistance. Selected isolates were characterised by auxotype and analysis was done for mutations within the quinolone resistance-determining region (QRDR) in the gyrA and parC genes, which confer fluoroquinolone resistance to the organism. Isolates with ciprofloxacin resistance increased significantly from 6.6% (1993-1994) to 73.5% (2002). The proportion of plasmid-mediated penicillin-resistant isolates (PPNG) decreased significantly from 7.9% (1993-1994) to 0.9% (2002). The percentage of chromosomal-mediated resistance to penicillin decreased from 27.4% in 2000 to 12.0% in 2001 but increased to 28.9% in 2002. The proportion of isolates with any type of resistance to tetracycline decreased from 24.7% in 2000 to 13.9% in 2001 and then increased to 22.3% in 2002. The proportion of prototrophic isolates significantly decreased from 84.4% in 1992-1993 to 7.7% in 2001, while that of the proline-requiring isolates significantly increased from 4.4% in 1992-1993 and 80.8% in 1998. The proline-requiring isolates were less susceptible to ciprofloxacin than the prototrophic or arginine-requiring isolates. Of 87 isolates resistant to ciprofloxacin, 2 (2.3%) contained five amino acid substitutions within the GyrA and ParC proteins, 76 (87.4%) contained three or four amino acid substitutions and 9 (10.3%) contained one or two amino acid substitutions. PMID- 15364301 TI - Mycoplasma genitalium urethritis in men. AB - Mycoplasma genitalium was first isolated from two men with non-gonococcal urethritis (NGU) and thereafter shown to produce urethritis in subhuman primates, inoculated intraurethrally. This mycoplasma has been detected significantly more often in patients with acute NGU, particularly in patients with non-chlamydial NGU, than in subjects without urethritis. The prevalence of M. genitalium positive non-chlamydial NGU ranges from 18 to 46% of all non-chlamydial NGU cases. In addition, the persistence of M. genitalium in the urethra after antimicrobial chemotherapy is associated with persistence or recurrence of NGU. The various results reported to date tend to support the proposition that M. genitalium is a pathogen of NGU. M. genitalium is highly susceptible to tetracyclines, macrolides, and some new fluoroquinolones, but the clinical data on the chemotherapy in M. genitalium-positive NGU is extremely limited. Because of the possible association between the post-treatment presence of M. genitalium in the urethra and persistent or recurrent NGU, the eradication of this mycoplasma from the urethra is essential in the management of patients with M. genitalium-positive NGU. Further studies are required to establish the optimal chemotherapy for M. genitalium-positive NGU. PMID- 15364302 TI - Bacterial resistance to antimicrobials in urinary isolates. AB - Escherichia coli accounted for about 80% of organisms in uncomplicated urinary tract infections (UTIs), followed by Staphylococcus spp. especially Staphylococcus saprophyticus, and Proteus mirabilis. Against E. coli isolates from patients with uncomplicated UTI, faropenem was the most effective. Up to 1999, fluoroquinolone-resistant isolates were not observed in patients with uncomplicated UTI, but in 2001 fluoroquinolone-resistant E. coli isolates emerged and accounted for about 8%. Various types of organisms were isolated in patients with complicated UTI. Enterococcus faecalis, E. coli, and Pseudomonas aeruginosa were the three most frequent organisms isolated. These three organisms accounted for 44.6%. Amongst oral agents, faropenem showed the lowest rate of resistance against E. coli followed by cephems. The rates of highly fluoroquinolone resistant and cefpodoxime-resistant E. coli isolates increased rapidly from 1998 to 2001. Fluoroquinolone-resistant P. aeruginosa isolates accounted for about 40% in 2001. Against this species, amikacin was the most effective antimicrobials among all agents tested. About 17% of Pseudomonas were resistant to carbapenem. Eight milligram per litre of ampicillin inhibited all E. faecalis isolates; about 60% of Enterococcus faecium were resistant to ampicillin. The rates of levofloxacin-resistant isolates of E. faecalis and E. faecium were 38 and 97% respectively. UTIs caused by vancomycin resistant enterococci (VRE) are rare in Japan. PMID- 15364303 TI - Urinary tract infections in African infants. AB - Urinary tract infection (UTI) causes significant illness in the first 2 years of life. The diagnosis in most developing countries is often overlooked due to the tedious nature of obtaining urine from these young ones who would not void voluntarily. Misdiagnosis very often lead to avoidable ill health and long-term renal damage. There is a need to diagnose UTI in febrile infants to alert clinicians. A prospective study of febrile infants aged up to 12 months on admission was undertaken in a 6-months period. Urine specimen was obtained by supra pubic aspiration and investigated. Out of 150 urine samples screened for UTI, 45 (30%) had positive bacterial growth. Escherichia coli (32%) and Proteus sp. (22%) formed more than 50% of the total isolates. The Gram positive bacteria isolated was Staphylococcus aureus representing 11%. All isolates were susceptible to cefuroxime and resistant to ampicillin. Susceptibility to amoxicillin/clavulanic acid was 77.8% and to nitrofurantoin was 67%. Only 11.1% of isolates were susceptible to cotrimoxazole. These results indicate that UTI is a common cause of pyrexia in infants requiring admission. These infants when clinically diagnosed as having UTI are treated with ampicillin and cotrimoxazole. They do not have the benefit of further investigation for renal abnormalities and subsequent antibiotic prophylactics to prevent recurrent infection, which could result in renal scarring and its associated problems later. PMID- 15364304 TI - Newer carbapenems for urinary tract infections. AB - Four carbapenems have been available clinically in Japan. These are imipenem/cilastatin (IMIP/CS) and panipenem/ betamipron (PANI/BP) of the older compounds and newer carbapenems such as biapem (BIAP) and meropenem (MERO). The latter compounds are relatively stable to dehydropeptidase-1 (DHP-1) and have been reported to have higher antimicrobial activities compared to the earlier carbapenems. The antimicrobial activity of these four carbapenems against fresh urinary isolates showed high activities against Enterobacteriacae such as Serratia marcescens, Enterobacter cloacae, Citrobacter freundii and Escherichia coli containing the class C-beta-lactamase- and extended spectrum beta-lactamase (ESBL)-producing strains compared to piperacillin (PIPC) and ceftazidime (CTAZ). Against Pseudomonas aeruginosa, the carbapenems, with the exception of panipenem showed strong antimicrobial activities compared to PIPC and CTAZ. High activities were also seen against Enterococcus faecalis, Staphylococcus aureus and Staphylococcus epidermidis, but methicillin-resistant strains were not affected. The first generation carbapenems showed better activity against E. faecalis than newer carbapenems. All four carbapenems were similar in clinical effectiveness in double blind trials for complicated urinary tract infections (UTIs). However, PANI/BP is less effective in UTIs caused by P. aeruginosa than IMIP/CS. MERO showed better eradication rate of P. aeruginosa than IMIP/CS. Retrospective analysis of treated cases using carbapenems showed a rapid defervescence in the treatment of febrile complicated UTIs, which were mainly caused by mixed infection of Gram-negative and Gram-positive bacteria, especially those involving P. aeruginosa and E. faecalis. PMID- 15364305 TI - New drugs for Gram-positive uropathogens. AB - Complicated urinary tract infections (UTIs) are frequent nosocomial infections. The bacterial spectrum encompasses Gram-negative but also Gram-positive pathogens in up to 30-40%. The existing treatment for Gram-positive pathogens is not always optimal. Antimicrobials for the treatment of Gram-positive uropathogens comprise older agents, such as aminopenicillins with or without beta-lactamase inhibitors and vancomycin, as well as newer fluoroquinolones, such as levofloxacin or gatifloxacin. However, resistant bacteria such as vancomycin-resistant enterococci (VRE) or methicillin-resistant Staphylococcus aureus (MRSA) (except vancomycin-resistant) are generally also not susceptible to the fluoroquinolones. Therefore new agents need to be assessed in the treatment of UTI. Daptomycin and linezolid are new antimicrobial agents with good efficacy against Gram-positive uropathogens as shown by their minimal inhibitory concentrations. In a phase II study the urinary bactericidal activity of linezolid versus ciprofloxacin in volunteers showed comparable activity of both drugs against fluoroquinolone susceptible Gram-positive uropathogens, whereas linezolid was also as active against fluoroquinolone resistant ones. The pharmacokinetics and the mode of action of these two antibiotics are discussed together with some clinical data in the context of therapeutic use in patients with complicated UTIs. PMID- 15364306 TI - Catheter-associated urinary tract infections: diagnosis and prophylaxis. AB - Catheter-associated urinary tract infections (CAUTI) are the commonest nosocomial infections worldwide. While they are often asymptomatic and frequently cost less than nosocomial surgical site infections or nosocomial pneumonia, they are major reservoirs of antimicrobial resistant pathogens. Numerous strategies have been devised in an attempt to reduce the incidence of CAUTI but few have proven effective. Novel technologies such as the potential use of antiseptic or antimicrobial coatings on catheters hold promise for possibly reducing these infections in the fight against antimicrobial resistance. PMID- 15364307 TI - NIDDK-sponsored chronic prostatitis collaborative research network (CPCRN) 5-year data and treatment guidelines for bacterial prostatitis. AB - Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a disabling condition that is poorly understood. The National Institutes of Diabetes and Digestive and Kidney Diseases-sponsored Collaborative Research Network has developed a symptom index, formed a cohort study, a case control study, a full scale randomised clinical trial, a resource utilisation study and clinical trials, as well as basic research studies, in an effort to better understand and manage patients with this condition. Cohort, case control and resource utilisation studies have confirmed the substantial impact of CP/CPPS. Clinical trials in basic research suggest that anti-inflammatory therapy and alpha-blocker therapy may be effective. The minority of patients with acute or chronic bacterial prostatitis continues to respond favourably to oral fluoroquinolone therapy. PMID- 15364308 TI - The role of unusual pathogens in prostatitis syndrome. AB - A total of 1442 patients with symptoms of chronic prostatitis were examined over a 4-year period at the Outpatient Department for Urogenital Infections, University Hospital for Infectious Diseases "Dr. Fran Mihaljevic", Zagreb, Croatia. An infectious aetiology was determined in 1070 (74.2%) patients. In 561 of 1070 (52.4%) patients the inflammatory finding (>10 WBC/hpf) was found in expressed prostatic secretions (EPS) or voided bladder urine (VB(3)). Normal, <10 WBCs/hpf was found in 362 of 536 (67.5%) patients with symptoms of chronic prostatitis in whom Chlamydia trachomatis was detected in EPS or VB(3), in 51 of 151 (33.8%) patients with isolated Trichomonas vaginalis and in 40 of 72 (55.6%) patients with isolated Ureaplasma urealyticum. Escherichia coli was the causative pathogen in 95, Enterococcus in 68, Proteus mirabilis in 37, Klebsiella pneumoniae in 16, Streptococcus agalactiae in 19, and Pseudomonas aeruginosa in 3 patients with chronic prostatitis. Other patients had a mixed infection. In patients with chronic bacterial prostatitis (CBP) caused by E. coli, P. mirabilis, K. pneumoniae, E. or S. agalactiae, an inflammatory finding was regularly found in EPS or VB(3). PMID- 15364309 TI - Infectious response to E. coli: molecular and genetic pathways. AB - Urinary tract infections are most commonly caused by type 1-piliated Escherichia coli (UPEC) and result in urothelial apoptosis, local cytokine release and neutrophil infiltration. A human urothelial cell line was incubated with various E. coli isolates and was then characterized by flow cytometry. UPEC induced rapid urothelial apoptosis that was dependent upon interactions mediated by type 1 pili. Laboratory isolates expressing type 1 pili-induced approximately 50% less apoptosis. UPEC blocked activity of a NF-kappaB-dependent reporter in response to inflammatory stimuli by stabilizing IkappaBalpha and UPEC rapidly altered cellular signalling pathways. Finally, blocking NF-kappaB activity increased the level of the laboratory strain-induced apoptosis to the level of apoptosis induced by UPEC. These results suggest that UPEC blocks NF-kappaB and enhances type 1 pili-induced apoptosis as a component of the uropathogenic programme. PMID- 15364310 TI - Antimicrobial susceptibility of uropathogens causing acute uncomplicated cystitis in female outpatients in South Korea: a multicentre study in 2002. AB - The objective of this study was to obtain data on the susceptibility of pathogens responsible for acute uncomplicated cystitis in South Korea to currently used antimicrobial agents. A total of 239 isolates were obtained from female outpatients with acute uncomplicated cystitis. The most prevalent causative organism was Escherichia coli (79.9%), followed by coagulase-negative staphylococci (CNS) (4.2%) and Enterobacteriaceae (8.2%). The mean rates of susceptibility to commonly prescribed drugs are ampicillin 35.5%, cefazolin 89.1%, ciprofloxacin 85.7%, and trimethoprim/sulphamethoxazole (TMP/SMX) 62.1%. Regionally, resistance to TMP/SMX among E. coli now exceeds 30% in all regions. The high prevalence of resistance among uropathogens requires on going surveillance to ensure the provision of effective empirical therapy for acute uncomplicated cystitis. PMID- 15364311 TI - A comparative multicentre study on the incidence of catheter-associated urinary tract infection between nitrofurazone-coated and silicone catheters. AB - The efficacy of nitrofurazone-coated urinary catheter in inhibitory activity of catheter-associated urinary tract infection (CAUTI) was evaluated. The incidence rate and onset of CAUTI after catheterisation of standard silicone urinary catheters and nitrofurazone-coated catheters was compared. There was no statistical significance between the two groups in the incidence rate of CAUTI. However, in patients who had indwelling urinary catheters for 5-7 days, the incidence rate of CAUTI was significantly lower in the experimental group. Logistic regression analysis showed that the two variables, including age and period of insertion, affected the incidence rate of CAUTI significantly. Nitrofurazone-coated catheters can be useful for inhibition of CAUTI in patients who have indwelling urinary catheter for 5-7 days and in old-age patients. PMID- 15364312 TI - Correlation between blood group phenotype and virulence properties of Escherichia coli in patients with chronic urinary tract infection. AB - A predisposition to urinary tract infection (UTI) is associated with the expression of P1 as well as the presence of ABO blood group antigen on the boundary layer and with the secretor state. Infectious microorganisms interfere with specific molecules on epithelial cells, these are antigens of the P and ABO blood group system. The blood group phenotype was examined in 53 women (age 42 +/ 12 years) with chronic non-obstructive UTI. The diagnosis was established on the basis of clinical history as well as clinical laboratory and radiological findings. The ABO phenotype and the P1 antigen were analysed by anti-A and anti-B as well as anti-P1 serum. The Lewis phenotyping was performed by incubating erythrocytes with anti-Le(a) and anti-Le(b) serum. In all patients, the blood group status were investigated. The proportion of persons with B-phenotype was 23% (the incidence of this feature in the German population is 14.5%). P1 antigen was found in 76% patients. In comparison with P1 antigen-negative individuals, P1 antigen-positive persons have a longer disease history and suffer more frequently from symptomatic events as well as destructive renal changes. The Le (a)-antigen was detected in 82% and the Le (b)-phenotype was observed in 18% of patients. The blood group phenotypes (ABO, Lewis and P1 antigens) represent an interesting natural aspect of local defence system against the invasive efforts of uropathogens. Antigen structures on uroepithelial cells for example, the glycolipids of the P antigen, serve as receptors for adhesion of microorganisms. PMID- 15364313 TI - Cardiac arrhythmias: the quest for a cure: a historical perspective. AB - During the last 40 years, much progress has been made in our understanding and management of cardiac arrhythmias. A major step in the late 1960s was to combine programmed electrical stimulation of the heart with intracardiac activation recording. This allowed: 1) localization of the site of the block in the atrioventricular conduction system in patients with bradycardia; and 2) identification of the site of origin and the mechanism of supraventricular and ventricular tachycardia. Combining information from intracardiac studies with findings on the 12-lead electrocardiogram (ECG) resulted in much better localization of conduction abnormalities and arrhythmias using the ECG. This new knowledge led to the development of new therapies, such as bradycardia and antitachycardia pacing, and surgery for supraventricular and ventricular tachycardia. A very important development in the treatment of life-threatening arrhythmias was the implantable defibrillator. Growing concern about failure to protect patients at risk for dying suddenly with antiarrhythmic drugs led to a rapid increase in their number. Cure by catheter ablation became possible for patients with different types of arrhythmias. Genetic analysis allowed the identification of different monogenic arrhythmic diseases. Several challenges remain: the epidemic of atrial fibrillation, arrhythmias in heart failure, and sudden death out-of-hospital. One-fifth of all deaths are sudden and unexpected. The important issue is how we are going to prevent these unnecessary deaths from occurring. PMID- 15364314 TI - Cardiovascular magnetic resonance imaging: current and emerging applications. AB - Magnetic resonance (MR) imaging is gaining importance in cardiology as the newest, most complex, and rapidly emerging noninvasive test of choice for patients with a multitude of cardiovascular problems. It has long been recognized to provide an accurate and reliable means of assessing the function and anatomy of the heart and great vessels, but its emerging role as one of the dominant imaging modalities in other aspects of cardiology such as perfusion imaging, atherosclerosis imaging, and coronary artery imaging cannot be understated. As MR technology evolves, newer therapeutic applications are also being developed, including specific MR-compatible catheters for electrophysiology studies/ablation as well as interventional cardiology related procedures, which may alter the way we practice cardiology in the future. Also, MR is entering an important phase in its evolution, with an anticipated exponential growth in its current applications and through the development of newer molecular imaging applications. It is anticipated that such developments will be coupled to the utilization of molecular markers to index biologic processes to allow for their in vivo visualization. This combination of biochemical markers and imaging methodology will also usher in an era of molecular imaging during which much progress in the diagnosis and treatment of cardiovascular disease is anticipated. PMID- 15364315 TI - International medical graduates and the cardiology workforce. AB - Recent publications have expressed the view that there is a shortage of cardiologists and it is growing worse. Both an increasing demand and a diminishing supply of cardiologists have been projected. An increase in the number of international medical graduates (IMGs) who enter cardiology practice has been proposed as a remedy for a projected shortage. The IMGs have to overcome challenges including clinical practice, language proficiency, and cultural differences before they are incorporated into the fabric of U.S. cardiology. With hard work, perseverance, excellence,compassionate care and support and mentoring, many have contributed to scientific and clinical cardiology in the U.S. Whether in the absence of a present crisis the projected shortage of cardiologists necessitates change in U.S. immigration policy is an open question. PMID- 15364316 TI - Stroke prevention with the angiotensin II type 1-receptor blocker candesartan in elderly patients with isolated systolic hypertension: the Study on Cognition and Prognosis in the Elderly (SCOPE). AB - OBJECTIVES: The aim of this study was to test the hypothesis that the angiotensin II type 1 receptor blocker (ARB) candesartan can reduce the risk of stroke in elderly patients with isolated systolic hypertension (ISH). BACKGROUND: Isolated systolic hypertension is the predominant form of hypertension in the elderly, and stroke is the most common cardiovascular (CV) complication. METHODS: In the Study on Cognition and Prognosis in the Elderly (SCOPE), 4,964 patients age 70 to 89 years were randomly assigned to double-blind candesartan or placebo with open label antihypertensive therapy (mostly thiazide diuretics) added as needed to control blood pressure. Of the 4,964 patients, 1,518 had ISH (systolic blood pressure >160 mm Hg and diastolic blood pressure <90 mm Hg). The present study is a predefined subgroup analysis of outcome results in the ISH patients. RESULTS: Of the ISH patients, 754 were randomized to the candesartan group and 764 to the control group. Over the study period, blood pressure was reduced by 22/6 mm Hg in the candesartan group and by 20/5 mm Hg in the control group (difference between treatments 2/1 mm Hg; p = 0.101 and 0.064). A total of 20 fatal/non-fatal strokes occurred in the candesartan group (7.2/1,000 patient-years) and 35 in the control group (12.5/1,000 patient-years); relative risk (RR) was 0.58 (95% confidence interval 0.33 to 1.00), that is, a RR reduction of 42% (p = 0.050 unadjusted, p = 0.049 adjusted for baseline risk). There were no marked or statistically significant differences between the treatment groups in other CV end points or all-cause mortality. CONCLUSIONS: In elderly patients with ISH, antihypertensive treatment based on the ARB candesartan resulted in a significant 42% RR reduction in stroke in comparison with other antihypertensive treatment, despite little difference in blood pressure reduction. PMID- 15364317 TI - Effects of a novel immune modulation therapy in patients with advanced chronic heart failure: results of a randomized, controlled, phase II trial. AB - OBJECTIVE: We sought to determine whether a novel, non-pharmacological form of immune modulation therapy (IMT), shown experimentally to reduce inflammatory and increase anti-inflammatory cytokines, improved outcomes in patients with advanced heart failure (HF). BACKGROUND: Immune activation contributes to the progression of HF, but treatments directed against inflammation have been largely unsuccessful. METHODS: Seventy-five HF patients (New York Heart Association [NYHA] functional class III to IV) were randomized to receive either IMT (n = 38) or placebo (n = 37) in a double-blind trial for six months, with continuation of standard HF therapy. Patients were evaluated using the 6-min walk test, changes in NYHA functional class, cardiac function, and quality of life assessments, as well as occurrence of death and hospitalization. RESULTS: There was no between group difference in 6-min walk test, but 15 IMT patients (compared with 9 placebo) improved NYHA functional classification by at least one class (p = 0.140). The Kaplan-Meier survival analysis showed that IMT significantly reduced the risk of death (p = 0.022) and hospitalization (p = 0.008). Analysis of a clinical composite score demonstrated a significant between-group difference (p = 0.006). There was no difference in left ventricular ejection fraction, but there was a trend toward improved quality of life (p = 0.110). CONCLUSIONS: These preliminary findings are consistent with the hypothesis that immune activation is important in the pathogenesis of HF and establish the basis for a phase III trial to define the benefit of IMT in chronic HF. PMID- 15364318 TI - Achieved platelet aggregation inhibition after different antiplatelet regimens during percutaneous coronary intervention for ST-segment elevation myocardial infarction. AB - OBJECTIVES: To evaluate the extent of platelet aggregation inhibition in patients with ST-segment elevation myocardial infarction (STEMI) undergoing percutaneous coronary intervention (PCI), treated with different antiplatelet agents and dosages. BACKGROUND: The extent of platelet aggregation inhibition is an independent predictor of major cardiac events after elective PCI. In STEMI patients undergoing PCI, routine dose of antiplatelet agents may be associated with less effective platelet aggregation inhibition. METHODS: Patients were treated with clopidogrel before angiography and randomized to abciximab, tirofiban, high-dose tirofiban, or no glycoprotein (GP) IIb/IIIa inhibitor; GP IIb/IIIa inhibitor bolus, followed by maintenance infusion, was administered after angiography, but before PCI. Platelet aggregation inhibition was assessed before angiography, immediately after PCI, and 1 and 6 h afterwards. RESULTS: The total study population consisted of 112 patients. Platelet aggregation inhibition was variable for individuals and suboptimal for all agents, particularly in the periprocedural period. Only with high-dose tirofiban, mean periprocedural platelet aggregation inhibition exceeded 80%. Angiographic parameters after PCI were not different between the groups. No relationship was found between the level of platelet aggregation and parameters of PCI success (Thrombolysis In Myocardial Infarction frame count and myocardial blush grade), after combining the data from all four groups studied. CONCLUSIONS: Platelet aggregation inhibition in STEMI patients undergoing PCI, treated with antiplatelet agents, is variable and suboptimal for all agents and dosages studied. Only with high-dose tirofiban, mean periprocedural platelet aggregation inhibition exceeded 80%. However, no relationship of platelet aggregation inhibition and angiographic outcome was found in this patient cohort. PMID- 15364319 TI - Lack of clopidogrel pretreatment effect on the relative efficacy of bivalirudin with provisional glycoprotein IIb/IIIa blockade compared to heparin with routine glycoprotein IIb/IIIa blockade: a REPLACE-2 substudy. AB - OBJECTIVES: The purpose of this study was to assess if clopidogrel pretreatment affects the relative efficacy of bivalirudin versus heparin with glycoprotein (GP) IIb/IIIa blockade for percutaneous coronary interventions (PCI). BACKGROUND: Although thienopyridine pretreatment may improve clinical outcomes with PCI, it is unknown if bivalirudin's efficacy compared with heparin is dependent upon such pretreatment. METHODS: The Randomized Evaluation in Percutaneous coronary intervention Linking Angiomax to reduced Clinical Events (REPLACE-2) trial was a double-blind, triple-dummy, randomized-controlled trial comparing heparin plus routine GP IIb/IIIa blockade (heparin group) with bivalirudin plus provisional GP IIb/IIIa blockade (bivalirudin group) during PCI. The primary end point was a composite of death, myocardial infarction (MI), urgent revascularization at 30 days, and major in-hospital bleeding. The secondary end point was a 30-day composite of death, MI, and urgent revascularization. Clopidogrel pretreatment was encouraged (300 mg loading, 75 mg/day). RESULTS: Of 6,010 patients enrolled, 5,893 received clopidogrel, with 85.8% in the bivalirudin and 84.6% in the heparin group receiving clopidogrel pretreatment. Bivalirudin (provisional GP IIb/IIIa blockade 7.2%) was noninferior to the heparin group for both primary and secondary end points. Clopidogrel pretreatment did not affect the relative efficacy of bivalirudin versus heparin with GP IIb/IIIa blockade, irrespective of pretreatment duration. Pretreatment was associated with significantly lower primary end point with bivalirudin (8.7% pretreatment vs. 12.9% no pretreatment, p = 0.007), and nonsignificantly with heparin (9.7% vs. 11.7%, respectively, p = 0.20). Multivariable models showed a trend toward lower primary and secondary end points with clopidogrel pretreatment. CONCLUSIONS: Clopidogrel pretreatment at the doses and time administered in this trial did not influence the relative efficacy of bivalirudin versus heparin plus GP IIb/IIIa blockade for PCI. However, pretreatment was associated with a trend towards lower clinical events after PCI. PMID- 15364320 TI - Vascular complications associated with arteriotomy closure devices in patients undergoing percutaneous coronary procedures: a meta-analysis. AB - OBJECTIVES: This study was designed to assess the safety of arteriotomy closure devices (ACDs) versus mechanical compression by meta-analysis in patients undergoing percutaneous transfemoral coronary procedures. BACKGROUND: Although ACDs are widely applied for hemostasis after percutaneous endovascular procedures, their safety is controversial. METHODS: Randomized, case-control, and cohort studies comparing access-related complications using ACDs versus mechanical compression were analyzed. The primary end point was the cumulative incidence of vascular complications, including pseudoaneurysm, arteriovenous fistula, retroperitoneal hematoma, femoral artery thrombosis, surgical vascular repair, access site infection, and blood transfusion. RESULTS: A total of 30 studies involving 37,066 patients were identified. No difference in complication incidence between Angio-Seal and mechanical compression was revealed in the diagnostic (Dx) setting (odds ratio [OR] 1.08, 95% confidence interval [CI] 0.11 to 10.0) or percutaneous coronary interventions (PCI) (OR 0.86, 95% CI 0.65 to 1.12). Meta-analysis of randomized trials only showed a trend toward less complications using Angio-Seal in a PCI setting (OR 0.46, 95% CI 0.20 to 1.04; p = 0.062). No differences were observed regarding Perclose in either Dx (OR 1.51, 95% CI 0.24 to 9.47) or PCI (OR 1.21, 95% CI 0.94 to 1.54) setting. An increased risk in complication rates using VasoSeal in the PCI setting (OR 2.25, 95% CI 1.07 to 4.71) was found. The overall analysis favored mechanical compression over ACD (OR 1.34, 95% CI 1.01 to 1.79). CONCLUSIONS: In the setting of Dx angiography, the risk of access-site-related complications was similar for ACD compared with mechanical compression. In the setting of PCI, the rate of complications appeared higher with VasoSeal. PMID- 15364321 TI - Differential mortality risk of postprocedural creatine kinase-MB elevation following successful versus unsuccessful stent procedures. AB - OBJECTIVES: This study was designed to evaluate the effect of periprocedural myocardial infarction (MI) on mortality according to success of the stent procedure. BACKGROUND: The mortality effect of periprocedural MI relative to successful versus unsuccessful procedures has not been examined. METHODS: All cause mortality during the first year was evaluated prospectively among 5,850 patients from coronary stent clinical trials. Myocardial infarction was classified according to creatine kinase-MB level as type 1 (>1 but <3 times normal), type 2 (>or=3 but 8 times normal or Q wave MI). Procedures were classified as successful unless there was a final diameter stenosis >50%; final Thrombolysis In Myocardial Infarction flow grade <3; final National Heart, Lung, and Blood Institute dissection grade >or=D; repeat revascularization within 24 h; or stent thrombosis within 24 h. RESULTS: Myocardial infarction was more frequent after unsuccessful procedures (69.6% vs. 20.4%, p < 0.001). Mortality during the first year was higher in patients with MI (2.8% vs. 1.7%, p = 0.01), but the effect was significant only for type 3 MI (4.7% vs. 1.7%, p = 0.008). Moreover, the mortality difference for any MI was confined to patients with unsuccessful procedures (13.1% vs. 0%, p = 0.03), with no significant effect among patients with otherwise successful procedures (2.1% vs. 1.7%, p > 0.20). The independent predictors of mortality were unsuccessful procedure (p < 0.001), diabetes mellitus (p = 0.001), history of prior MI (p = 0.003), multivessel disease (p = 0.006), and advancing age (p < 0.001), but not periprocedural MI. CONCLUSIONS: The association of periprocedural MI with increased mortality during the first year following stent placement was confined to patients with unsuccessful procedures. PMID- 15364322 TI - Prognostic utility of comparative methods for assessment of ST-segment resolution after primary angioplasty for acute myocardial infarction: the Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications (CADILLAC) trial. AB - OBJECTIVE: This study was done to assess and compare the prognostic significance of multiple methods for measuring ST-segment elevation resolution (STR) following primary percutaneous coronary intervention (PCI). BACKGROUND: Resolution of ST segment elevation (STE) is a powerful predictor of both infarct-related artery patency and mortality in acute myocardial infarction (AMI). Recent thrombolytic studies have suggested that simple measures of STR may be as powerful as more complex algorithms. The optimal method of assessing STR following primary PCI has not been studied. METHODS: We analyzed 700 patients with technically adequate baseline and post-PCI electrocardiograms from the Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications (CADILLAC) trial. Five methods were used to assess STR: 1) summed %STR across multiple leads (SigmaSTR); 2) %STR in the single lead with maximum baseline STE (MaxSTR); 3) absolute maximum STE before the procedure; 4) absolute maximum STE after intervention (MaxSTPost); and 5) a categorical variable based upon MaxSTPost (High Risk). RESULTS: At 30 days, SigmaSTR, MaxSTR, and MaxSTPost all correlated strongly with mortality (p = 0.004, p = 0.005, and p < 0.0001, respectively) and the combined end point of mortality or reinfarction (p = 0.001, p = 0.001, and p < 0.0001). At one year, SigmaSTR and MaxSTPost correlated with mortality (p = 0.04, p = 0.0001), reinfarction (p = 0.02, p = 0.0015), and the combined end point (p = 0.02, p < 0.0001). By multivariate analysis, only the simpler measures of MaxSTPost and High Risk categorization independently predicted all outcomes at both time points. CONCLUSIONS: The STR following primary PCI in AMI correlates strongly with mortality and reinfarction, independent of target vessel patency. The simple measure of the maximal residual degree of STE after primary PCI is a strong independent predictor of both survival and freedom from reinfarction at 30 days and 1 year. PMID- 15364323 TI - Noninvasive visualization of coronary artery bypass grafts using 16-detector row computed tomography. AB - OBJECTIVES: The aim of this study was to evaluate the diagnostic accuracy of a 16 detector row computed tomography (CT) scanner for the assessment of coronary artery bypass grafts. BACKGROUND: A new generation of multislice spiral CT scanners, equipped with more and thinner detector rows, allows for reliable noninvasive detection of obstructive coronary artery disease. METHODS: The study included 51 consecutive patients. Three patients had to be excluded from the study due to arrhythmias or fast heart rates despite beta-blockade. A total of 48 patients with 131 coronary artery bypass grafts (internal mammary artery, n = 40; venous grafts, n = 91) were examined by computed tomography angiography (CTA) and by invasive coronary angiography (ICA) using a 16-detector row CT scanner. For cardiac protocols, only the 12 inner detector rings are applied. All CT examinations were performed with retrospective electrocardiogram gating at a mean heart rate of 64 +/- 5 beats/min; 120 ml of Xenetix 300 (Guerbert GmbH, Sulzbach, Germany) were continuously injected. The bypass graft patency and the presence of stenoses as well as the proximal and distal anastomoses were evaluated by two experienced readers. RESULTS: All bypass grafts and 74% of the distal bypass anastomoses could be visualized by CTA; 21 bypass graft occlusions and 1 significant stenosis were detected by CTA and confirmed by ICA. Five false positive and one false negative finding resulted in a sensitivity of 96%, a specificity of 95%, a positive predictive value of 81%, and a negative predictive value of 99%. CONCLUSIONS: Sixteen-detector row CT scanner technology allows for the reliable visualization of coronary bypass grafts. Dysfunctional bypass grafts can be detected with high diagnostic accuracy. This technology can be used as a noninvasive test for patients with suspected graft dysfunction. PMID- 15364324 TI - Noninvasive detection of coronary lesions using 16-detector multislice spiral computed tomography technology: initial clinical results. AB - OBJECTIVES: The aim of our study was to evaluate the feasibility of detecting coronary artery lesions using a new computed tomography (CT) scanner with 16 detectors and faster gantry rotation. BACKGROUND: Computed tomography angiography of the coronaries permits assessment of the coronaries; however, image quality is still impaired by motion artifacts and calcifications. METHODS: Sixty patients scheduled for conventional coronary angiography (CCA) were additionally studied by multislice spiral computed tomography (MSCT). Calcium scores and a contrast enhanced visualization of the coronaries were performed and analyzed regarding evaluability, presence of coronary artery lesions, and correct clinical diagnosis. RESULTS: Calcium scoring was successful in all patients; 58 of 60 patients had a diagnostic contrast-enhanced scan. Mean calcium score was 506 +/- 743 Agatston score equivalent (ASE); 13 of 58 (22%) patients had an ASE >or=1,000, 46 of 58 (78%) patients <1,000. In 763 coronary segments, CCA detected a total of 75 lesions >or=50%. The MSCT correctly assessed 54 of these. Twenty one lesions were missed or incorrectly underestimated. Sensitivity was 72%, specificity 97%. When restricting analysis to patients with an ASE <1,000, 40 significant lesions >or=50% were seen on CCA, and MSCT correctly detected 39 lesions (sensitivity 98%, specificity 98%). Regardless of any threshold, the correct clinical diagnosis could be obtained in 58 of 60 (97%) of all patients. CONCLUSIONS: In individuals with low-to-moderate amounts of coronary artery calcium, 16-detector CT coronary angiography has high sensitivity and specificity for the diagnosis of significant coronary artery stenosis. PMID- 15364325 TI - Noninvasive multi-slice computed tomography coronary angiography: an emerging clinical modality. PMID- 15364326 TI - Should coronary artery bypass graft surgery patients with mild or moderate aortic stenosis undergo concomitant aortic valve replacement? A decision analysis approach to the surgical dilemma. AB - OBJECTIVES: This study utilizes Markov decision analysis to assess the relative benefits of prophylactic aortic valve replacement (AVR) at the time of coronary artery bypass graft surgery (CABG). Multiple sensitivity analyses were also performed to determine the variables that most profoundly affect outcome. BACKGROUND: The decision to perform CABG or concomitant CABG and AVR (CABG/AVR) in asymptomatic patients who need CABG surgery but have mild to moderate aortic stenosis (AS) is not clear-cut. METHODS: We performed Markov decision analysis comparing long-term, quality-adjusted life outcomes of patients with mild to moderate AS undergoing CABG versus CABG/AVR. Age-specific morbidity and mortality risks with CABG, CABG/AVR, and AVR after a prior CABG were based on the Society of Thoracic Surgeons national database (n = 1,344,100). Probabilities of progression to symptomatic AS, valve-related morbidity, and age-adjusted mortality rates were obtained from available published reports. RESULTS: For average AS progression, the decision to replace the aortic valve at the time of elective CABG should be based on patient age and severity of AS measured by echocardiography. For patients under age 70 years, an AVR for mild AS is preferred if the peak valve gradient is >25 to 30 mm Hg. For older patients, the threshold increases by 1 to 2 mm Hg/year, so that an 85-year-old patient undergoing CABG should have AVR only if the gradient exceeds 50 mm Hg. The AS progression rate also influences outcomes. With slow progression (<3 mm Hg/year), CABG is favored for all patients with AS gradients <50 mm Hg; with rapid progression (>10 mm Hg/year), CABG/AVR is favored except for patients >80 years old with a valve gradient <25 mm Hg. CONCLUSIONS: This study provides a decision aid for treating patients with mild to moderate AS requiring CABG surgery. Predictors of AS progression in individual patients need to be better defined. PMID- 15364327 TI - Clinical prediction rule for atrial fibrillation after coronary artery bypass grafting. AB - OBJECTIVES: This study was designed to devise and validate a practical prediction rule for atrial fibrillation/atrial flutter (AF) after coronary artery bypass grafting (CABG) using easily available clinical and standard electrocardiographic (ECG) criteria. BACKGROUND: Reported prediction rules for postoperative AF have suffered from inconsistent results and controversy surrounding the added predictive value of a prolonged P-wave duration. METHODS: In 1,851 consecutive patients undergoing CABG with cardiopulmonary bypass, preoperative clinical characteristics and standard 12-lead ECG data were examined. Patients were continuously monitored for the occurrence of sustained postoperative AF while hospitalized. Multiple logistic regression was used to determine significant predictors of AF and to develop a prediction rule that was evaluated through jackknifing. RESULTS: Atrial fibrillation occurred in 508 of 1,553 patients (33%). Multivariate analysis showed that greater age (odds ratio [OR] 1.1 per year [95% confidence intervals (CI) 1.0 to 1.1], p < 0.0001), prior history of AF (OR 3.7 [95% CI 2.3 to 6.0], p < 0.0001), P-wave duration >110 ms (OR 1.3 [95% CI 1.1 to 1.7], p = 0.02), and postoperative low cardiac output (OR 3.0 [95% CI 1.7 to 5.2], p = 0.0001) were independently associated with AF risk. Using the prediction rule we defined three risk categories for AF: <60 points, 61 of 446 (14%); 60 to 79 points, 330 of 908 (36%); and >or=80 points, 117 of 199 (59%). The area under the receiver-operator characteristic curve for the model was 0.69. CONCLUSIONS: These data show that post-CABG AF can be predicted with moderate accuracy using easily available patient characteristics and may prove useful in prognostic and risk stratification of patients after CABG. The presence of intraatrial conduction delay on ECG contributed least to the prediction model. PMID- 15364328 TI - Reduced incidence of hypertension after heterotopic cardiac transplantation compared with orthotopic cardiac transplantation: evidence that excision of the native heart contributes to post-transplant hypertension. AB - OBJECTIVES: This study was designed to test the hypothesis that heterotopic heart transplant (HHT) patients have lower blood pressure than orthotopic cardiac transplant (OCT) patients because their native heart is involved in blood pressure homeostasis. BACKGROUND: Hypertension occurs more frequently after OCT than after liver or lung transplantation, suggesting that transplantation of the heart itself contributes to post-transplant hypertension. METHODS: Blood pressure and related measurements in 233 OCT and 38 HHT patients were studied retrospectively post-transplant. RESULTS: Systolic blood pressure (SBP) was persistently lower among HHT patients (means 121 vs. 137, 126 vs. 137, 125 vs. 139, and 128 vs. 143 mm Hg at month 3 and years 1, 3, and 5 respectively, p < 0.005). Left ventricular and aortic systolic pressures were also lower (130 vs. 143 mm Hg, p = 0.01 and 129 vs. 142 mm Hg, p = 0.01). Multivariable analysis with age, gender, body mass index, creatinine, steroids, cyclosporine, use of antihypertensive medication, donor left ventricular ejection fraction, donor weight, and type of transplant as covariables showed HHT to be independently associated with a lower SBP at each time point (beta-coefficients -16.2, -12.1, 13.3, and -14.2 mm Hg, p < 0.01). The adjusted hazard ratio for the development of systolic hypertension among HHT compared with OCT patients was 0.59 (95% confidence interval 0.39 to 0.91, p = 0.017). CONCLUSIONS: Heterotopic heart transplant patients had lower SBP than OCT patients, consistent with the hypothesis that the native heart continues to contribute to blood pressure homeostasis. PMID- 15364329 TI - Increased incidence of life-threatening ventricular arrhythmias in implantable defibrillator patients after the World Trade Center attack. AB - OBJECTIVES: This study was designed to evaluate whether the destruction of the World Trade Center (WTC) on September 11, 2001 (9/11), led to an increased frequency of ventricular arrhythmias among patients fitted with an implantable cardioverter-defibrillator (ICD). BACKGROUND: The WTC attack induced psychological distress. Because ICDs store all serious arrhythmias for months, the attack provided a unique opportunity to compare pre- and post-9/11 frequencies of potentially lethal arrhythmias among ICD patients. METHODS: Two hundred consecutive ICD patients who presented for regularly scheduled follow-up to six affiliated clinics were recruited into this observational study. The electrograms stored in the ICDs for the three months before 9/11 and 13 months thereafter were scrutinized in a blinded manner (relative to date) for all ventricular tachyarrhythmias (tachycardia or fibrillation) triggering ICD therapy. RESULTS: The frequency of tachyarrhythmias increased significantly for the 30 days post-9/11 (p = 0.004) relative to all other months between May 2001 and October 2002. In the 30 days post-9/11, 16 patients (8%) demonstrated tachyarrhythmias, compared with only seven (3.5%) in the preceding 30 days, representing a 2.3-fold increase in risk (95% confidence interval 1.1 to 4.9; p = 0.03). The first arrhythmic event did not occur for three days following 9/11, with events accumulating in a progressive non-clustered pattern. CONCLUSIONS: Ventricular arrhythmias increased by more than twofold among ICD patients following the WTC attack. The delay in onset and the non-clustered pattern of these events differ sharply from effects following other disasters, suggesting that subacute stress may have served to promote this arrhythmogenesis. PMID- 15364330 TI - The World Trade Center attack: increased frequency of defibrillator shocks for ventricular arrhythmias in patients living remotely from New York City. AB - OBJECTIVES: The goal of this study was to determine whether the World Trade Center (WTC) attack on September 11, 2001, had an effect on the occurrence of ventricular arrhythmias among patients with implantable cardioverter defibrillators (ICDs) living in Florida. BACKGROUND: Increased frequencies of ICD therapies for ventricular arrhythmias were reported among patients in New York City after the attack on the WTC. Whether this effect also occurred in patients living geographically distant from New York is unknown. METHODS: This was an observational study consisting of 132 ICD patients who presented to the University of Florida and the Veterans Affairs Medical Center in Gainesville, Florida, for routine ICD follow-up around the time of the WTC attack. The occurrence of ventricular tachyarrhythmias triggering ICD therapy in the 30 days before and after September 11 constituted the primary end point. RESULTS: In the 30 days following the WTC attack, a total of 14 patients (11%) had ventricular tachyarrhythmias, compared with 5 (3.8%) in the preceding 30 days (p = 0.0389, 95% confidence interval [CI] 0.4 to 13.3). This represents a 2.8-fold risk increase. Patients with ventricular arrhythmias both before and after September 11 demonstrated a rate increase of 2.38 events per patient (p = 0.0231, 95% CI 1.03 to 13.97). CONCLUSIONS: The frequency of ventricular arrhythmias requiring ICD treatment increased by 68% among patients in Florida around the WTC attack. These findings suggest that stress-related arrhythmogenesis due to the WTC tragedy was not restricted to the geographic location of the attack. A major national tragedy may cause widespread increased risk of potentially life threatening ventricular arrhythmias. PMID- 15364331 TI - Current burden of sudden cardiac death: multiple source surveillance versus retrospective death certificate-based review in a large U.S. community. AB - OBJECTIVES: We sought to determine the annual incidence of sudden cardiac death (SCD) in the general population using a prospective approach. To assess the validity of retrospective surveillance, a simultaneous comparison was made with a death certificate-based method of determining SCD incidence. BACKGROUND: Accurate surveillance and characterization of SCD in the general population is likely to significantly facilitate current and future community-based preventive and therapeutic interventions. METHODS: We performed a prospective evaluation of SCD among all residents of Multnomah County, Oregon (population 660,486) using multiple sources of surveillance. A comprehensive analysis of circumstances of death, medical records, and available autopsy data was performed. Comparisons were made with a retrospective, death certificate-based determination of SCD incidence using International Classification of Diseases-Version 10 codes and location of death. RESULTS: Between February 1, 2002, and January 31, 2003, 353 residents suffered SCD (incidence 53 of 100,000; median age 69 years, 57% male) accounting for 5.6% of overall mortality. Of these, 75 cases (21%) were identified using sources other than first responders. Resuscitation was attempted in 237 cases (67%) and successful (survival to hospital discharge) in 28 (8%). The retrospective death certificate-based review yielded 1,007 cases (incidence 153 of 100,000; median age 81 years, 51% male), and the positive predictive value of this methodology was 19%. CONCLUSIONS: Sudden cardiac death accounts for 5.6% of annual mortality, and prospective evaluation in the general population appears to be feasible. The use of multiple sources of ascertainment and information significantly enhances phenotyping of SCD cases. Retrospective death certificate based surveillance results in significant overestimation of SCD incidence. PMID- 15364332 TI - Risk of proarrhythmic events in the Atrial Fibrillation Follow-up Investigation of Rhythm Management (AFFIRM) study: a multivariate analysis. AB - OBJECTIVES: This study examined the risk of proarrhythmic events in patients receiving antiarrhythmic drugs for treatment of atrial fibrillation (AF) according to present-day safety guidelines. BACKGROUND: Advances in understanding the proarrhythmic risk of antiarrhythmic drugs has led to development of safety guidelines for these agents. Such guidelines were used in the Atrial Fibrillation Follow-up Investigation of Rhythm Management (AFFIRM) study. METHODS: This study was an analysis of the risk of arrhythmic events (arrhythmic death, resuscitated cardiac arrest, sustained ventricular tachycardia (VT), and torsade de pointes VT) in the antiarrhythmic drug arm of the AFFIRM study. Each time an antiarrhythmic drug was begun, it was counted as an exposure to that drug and the risk of an arrhythmic event was calculated. RESULTS: A total of 2,033 patients received 3,030 exposures to antiarrhythmic drugs. Ninety-six arrhythmic events occurred by six years. Patients with a left ventricular ejection fraction <40% had more arrhythmic events. Twelve documented cases of torsade de pointes VT were noted. The incidence of torsade de pointes was 0.6% at five years (95% confidence interval 0.32 to 1.07). CONCLUSIONS: The overall risk of adverse arrhythmic events upon exposure to antiarrhythmic drugs in the AFFIRM study was reasonably low. Strict criteria for the safe use of antiarrhythmic drugs were successful in minimizing proarrhythmic events. PMID- 15364333 TI - An intronic mutation causes long QT syndrome. AB - OBJECTIVES: The purpose of this research was to determine whether an intronic variant (T1945+6C) in KCNH2 is a disease-causing mutation, and if expanded phenotyping criteria produce improved identification of long QT syndrome (LQTS) patients. BACKGROUND: Long QT syndrome is usually caused by mutations in conserved coding regions or invariant splice sites, yet no mutation is found in 30% to 50% of families. In one such family, we identified an intronic variant in KCNH2. Long QT syndrome diagnosis is hindered by reduced penetrance, as the long QT phenotype is absent on baseline electrocardiogram (ECG) in about 30%. METHODS: Fifty-two family members were phenotyped by baseline QTc, QTc maximum on serial ECGs (Ser QTc-max), and on exercise ECGs (Ex QTc-max) and by T-wave patterns. Linkage analysis tested association of the intronic change with phenotype. The consequences of T1945+6C on splicing was studied using a minigene system and on function by heterologous expression. RESULTS: Expanded phenotype/pedigree criteria identified 23 affected and 29 unaffected. Affected versus unaffected had baseline QTc 484 +/- 48 ms versus 422 +/- 20 ms, Ser QTc-max 508 +/- 48 ms versus 448 +/- 10 ms, Ex QTc-max 513 +/- 54 ms versus 444 +/- 11 ms, and LQT2 T waves in 87% versus 0%. Linkage analysis demonstrated a logarithm of odds score of 10.22. Splicing assay showed T1945+6C caused downstream intron retention. Complementary deoxyribonucleic acid with retained intron 7 failed to produce functional channels. CONCLUSIONS: T1945+6C is a disease-causing mutation. It alters KCNH2 splicing and cosegregates with the LQT2 phenotype. Expanded ECG criteria plus pedigree analysis provided accurate clinical diagnosis of all carriers including those with reduced penetrance. Intronic mutations may be responsible for LQTS in some families with otherwise negative mutation screening. PMID- 15364334 TI - Serum levels of interleukin-10 on admission as a prognostic predictor of human fulminant myocarditis. AB - OBJECTIVES: We assessed the significance of serum cytokine levels in patients with fulminant myocarditis. BACKGROUND: Although many investigations have demonstrated the crucial role of cytokines in the development of myocarditis, it remains uncertain whether serum levels of cytokines enable one to predict the prognosis of human myocarditis, especially concerning cardiogenic shock (CS) requiring a mechanical cardiopulmonary support system (MCSS). METHODS: We studied 22 consecutive patients with fulminant myocarditis and compared them with 15 patients with acute myocardial infarction (AMI) requiring MCSS. The patients with myocarditis were classified into three groups: eight patients with CS requiring MCSS on admission (group 1); six patients who unexpectedly lapsed into CS requiring MCSS more than two days after catecholamine had been initiated (group 2); and eight patients without MCSS (group 3). Furthermore, 14 patients with myocarditis requiring MCSS were divided into a fatal group (n = 5) and a survival group (n = 9). Biochemical markers, including serum cytokine levels and hemodynamic variables on admission, were analyzed. RESULTS: Serum levels of interleukin (IL)-10 and tumor necrosis factor-alpha, but not other cytokines, were significantly higher in myocarditis than in AMI. Only serum levels of IL-10 were significantly higher in group 1 and 2 than in group 3 (49.1 +/- 37.5/20.7 +/ 17.6 pg/ml vs. 2.4 +/- 1.1 pg/ml; p = 0.0008/0.0012). Serum IL-10 levels were also significantly higher in the fatal group than in the survival group with myocarditis (74.0 +/- 27.0 pg/ml vs. 16.4 +/- 8.8 pg/ml; p = 0.003). CONCLUSIONS: Serum IL-10 levels on admission enabled one to predict subsequent CS requiring MCSS and mortality of fulminant myocarditis patients. PMID- 15364335 TI - Interleukin-10: biomarker or pathologic cytokine in fulminant myocarditis? PMID- 15364336 TI - Furosemide and the progression of left ventricular dysfunction in experimental heart failure. AB - OBJECTIVES: We tested the hypothesis that furosemide accelerates the progression of left ventricular systolic dysfunction in a tachycardia-induced porcine model of heart failure. BACKGROUND: Furosemide activates the renin-angiotensin aldosterone system in patients with congestive heart failure (CHF). Such activation may contribute to CHF progression, but prospective data are lacking. METHODS: Thirty-two Yorkshire pigs were randomized to furosemide (1 mg/kg intramuscularly daily, mean 16.1 +/- 0.9 mg) or placebo. Thereafter, a pacing model of heart failure was utilized to produce systolic dysfunction in both sets of animals (fractional shortening <0.16 by echocardiogram). The goal was to determine if furosemide would accelerate the progression of left ventricular dysfunction in the "treated" group. After sacrifice, sodium-calcium exchanger currents and their responsiveness to isoproterenol were measured during voltage clamp. All investigators were blinded to treatment assignment. RESULTS: Furosemide shortened the time to left ventricular dysfunction (35.1 +/- 5.1 days in placebo versus 21.4 +/- 3.2 days for furosemide animals; p = 0.038, log-rank test). By day 14, aldosterone levels were significantly higher in furosemide animals (43.0 +/- 11.8 ng/dl vs. 17.6 +/- 4.5 ng/dl; p < 0.05). Serum sodium was reduced (133.0 +/- 0.9 mmol/l furosemide vs. 135.7 +/- 0.8 mmol/l placebo; p < 0.05), but no difference in norepinephrine, potassium, magnesium, creatinine, or urea nitrogen was present. Basal sodium-calcium exchanger currents were significantly increased and isoproterenol responsiveness depressed by furosemide. CONCLUSIONS: Tachycardic pigs given furosemide had significant acceleration of both contractile and metabolic features of CHF, including left ventricular systolic dysfunction, elevated serum aldosterone levels, and altered calcium handling in a controlled experimental model of heart failure. PMID- 15364337 TI - Furosemide in the long-term management of heart failure: the good, the bad, and the uncertain. PMID- 15364338 TI - Rapid reversal of endothelial dysfunction in hypercholesterolemic apolipoprotein E-null mice by recombinant apolipoprotein A-I(Milano)-phospholipid complex. AB - OBJECTIVES: In this study, we examined whether a reconstituted high-density lipoprotein (HDL) utilizing recombinant apolipoprotein A-I(Milano) (apo A I(M))/phospholipid complex (PC) could restore normal endothelial function in hypercholesterolemic apolipoprotein (apo) E-null mice. BACKGROUND: We have previously shown antiatherosclerotic and vasculoprotective effects of recombinant apo A-I(M). METHODS: A perfused vessel preparation was used to examine vascular responses in control wild-type, untreated, and treated apo E-null mice. Aortic tissue cholesterol content and platelet aggregation were also measured. RESULTS: Endothelium-dependent vasodilator responses to acetycholine were significantly inhibited in untreated apo E-null mice compared with control wild-type mice (p < 0.001). Treatment of the mice for five weeks with once every-other-day intravenous bolus injections of apo A-I(M)/PC restored endothelium-dependent dilation in a dose-dependent manner (p < 0.01 at 80 mg/kg dose). The improvement in endothelial function was associated with a reduction in aortic cholesterol content and reduced platelet aggregability and occurred despite severe and persistent hypercholesterolemia. Neither treatment with free protein nor phospholipid carrier alone produced any significant effects. We performed additional experiments in vitro in isolated rabbit carotid arteries to compare the effects on lysophosphatidylcholine (LPC)-induced endothelial dysfunction. Treatment with apo A-I(M)/PC prevented impairment of endothelium-dependent vasodilator responses to acetylcholine to a greater degree than either wild-type apo A-I or plasma-derived HDL. CONCLUSIONS: Our results indicate a rapid improvement in endothelial dysfunction with recombinant apo A-I(M)/PC that is associated with mobilization of tissue cholesterol. Taken together with previously established antiatherosclerotic and antithrombotic effects, these findings suggest significant vasculoprotective effects with apo A-I(M)/PC therapy. PMID- 15364339 TI - Physical inactivity causes endothelial dysfunction in healthy young mice. AB - OBJECTIVES: We sought to determine if physical inactivity affects endothelial function in young healthy individuals. BACKGROUND: Recent studies have linked exercise training to increased bioavailability of vascular nitric oxide (NO) and to improved endothelial function in patients with cardiovascular disorders. The effects of physical inactivity on normal vascular endothelial function are not known. METHODS: Healthy young male C57Bl/6 mice living in groups of five in large cages, where they were running, climbing, and fighting during their active cycle, were randomly assigned to stay there or to live alone in small cages where they were predominantly resting. After five and nine weeks citrate synthase activity (a measure of mitochondrial respiratory chain activity), heart weight/body weight ratio, vascular reactivity, and protein expression of endothelial nitric oxide synthase (eNOS) were assessed. RESULTS: Singularized mice showed a reduction of citrate synthase activity (p < 0.05), of endothelium-dependent vasorelaxation (to 65 +/- 5% of control levels; p < 0.001), and of eNOS protein expression (to 53 +/ 8% of control levels; p < 0.01). In striking contrast, vascular responses to potassium chloride, phenylephrine, and the NO-donor racemic S-nitroso-N acetylpenicillamine were unchanged. The alterations of vascular eNOS-activity were completely reversible when singularized mice underwent exercise. In mice living in groups, exercise showed only a small effect on aortic eNOS expression. CONCLUSIONS: In young healthy individuals physical inactivity induces endothelial dysfunction, which is completely reversible by a short period of moderate exercise training. We suggest that physical inactivity, the so-called sedentary lifestyle, increases cardiovascular risk in young healthy individuals by inducing endothelial dysfunction. PMID- 15364340 TI - Primary results of the Rapid Emergency Department Heart Failure Outpatient Trial (REDHOT). A multicenter study of B-type natriuretic peptide levels, emergency department decision making, and outcomes in patients presenting with shortness of breath. AB - OBJECTIVES: The purpose of this study was to examine the relationships among B type natriuretic peptide (BNP) levels within the diagnostic range, perceived congestive heart failure (CHF) severity, clinical decision making, and outcomes of the CHF patients presenting to emergency department (ED). BACKGROUND: Since BNP correlates with the presence of CHF, disease severity, and prognosis, we hypothesized that BNP levels in the diagnostic range offer value independent of physician decision making with regard to critical outcomes in emergency medicine. METHODS: The Rapid Emergency Department Heart failure Outpatient Trial (REDHOT) study was a 10-center trial in which patients seen in the ED with shortness of breath were consented to have BNP levels drawn on arrival. Entrance criteria included a BNP level >100 pg/ml. Physicians were blinded to the actual BNP level and subsequent BNP measurements. Patients were followed up for 90 days after discharge. RESULTS: Of the 464 patients, 90% were hospitalized. Two-thirds of patients were perceived to be New York Heart Association (NYHA) functional class III or IV. The BNP levels did not differ significantly between patients who were discharged home from the ED and those admitted (976 vs. 766, p = 0.6). Using logistic regression analysis, an ED doctor's intention to admit or discharge a patient had no influence on 90-day outcomes, while the BNP level was a strong predictor of 90-day outcome. Of admitted patients, 11% had BNP levels <200 pg/ml (66% of which were perceived NYHA functional class III or IV). The 90-day combined event rate (CHF visits or admissions and mortality) in the group of patients admitted with BNP <200 pg/ml and >200 pg/ml was 9% and 29%, respectively (p = 0.006). CONCLUSIONS: In patients presenting to the ED with heart failure, there is a disconnect between the perceived severity of CHF by ED physicians and severity as determined by BNP levels. The BNP levels can predict future outcomes and thus may aid physicians in making triage decisions about whether to admit or discharge patients. Emerging clinical data will help further refine biomarker guided outpatient therapeutic and monitoring strategies involving BNP. PMID- 15364341 TI - Heart failure in the emergency department: is B-type natriuretic peptide a better prognostic indicator than clinical assessment? PMID- 15364342 TI - Patient safety: a(nother) call for action. PMID- 15364343 TI - More on the history of arrhythmia in long QT syndrome. PMID- 15364344 TI - Impaired vascular reactivity in patients with erectile dysfunction. PMID- 15364346 TI - Exercise release of cardiac natriuretic peptides is markedly enhanced when patients with coronary artery disease are treated medically by beta-blockers. PMID- 15364348 TI - Article retraction: Biotransformation of drugs by microbial cultures for predicting drug metabolism. PMID- 15364349 TI - ORC-associated replication factors as biomarkers for cancer. AB - Early detection and treatment of cancer are of central importance to improving patient prognoses. Traditional biomarkers of cell proliferation, such as Ki-67 and PCNA, have had a mixed clinical track record, proving to be good indicators of certain types of cancers but of limited use for many others. Recently, human counterparts of replication factors originally identified in budding yeast have shown great promise as new cancer biomarkers. Each of these factors has been shown to interact with the origin recognition complex (ORC) in yeast, and each has an essential role in the initiation of DNA replication. Studies with minichromosome maintenance (MCM) family proteins show that their levels are upregulated in tumor cells and are much better indicators of a wide variety of cancers than traditional biomarkers. Similarly encouraging results have been obtained in preliminary studies examining Cdc6 protein and Cdc7 kinase transcript levels in normal and cancerous cells. PMID- 15364350 TI - Immobilized viable microbial cells: from the process to the proteome em leader or the cart before the horse. AB - Biotechnological processes based on immobilized viable cells have developed rapidly over the last 30 years. For a long time, basic studies of the physiological behaviour of immobilized cells (IC) have remained in the shadow of the applications. Natural IC structures, i.e. biofilms, are being increasingly investigated at the cellular level owing to their definite importance for human health and in various areas of industrial and environmental relevance. This review illustrates this paradoxical development of research on ICs, starting from the initial rationale for IC emergence and main application fields of the technology--with particular emphasis on those that exploit the extraordinary resistance of ICs to antimicrobial compounds--to recent advances in the proteomic approach of IC physiology. PMID- 15364351 TI - Tackling the case report. AB - OBJECTIVE: Resuscitative measures are required to ensure the survival of the case report in medical writing. The aim of this study was to assess a series of case reports for quality improvement. METHODS: All articles pertaining to a single patient published in Auris Nasus Larynx and The Journal of Laryngology and Otology from January to December 2000 inclusive were examined. Data recorded included authorship numbers and origin of the report, pertinence of the accompanying abstract, length of the article, claims of a first and the number of references included in the bibliography. RESULTS: Twelve different countries produced 129 papers in the two journals. Author numbers ranged from 2 to 7 (mean 4.56) in Auris Nasus Larynx, and were 1 to 11 (mean 2.63) in The Journal of Laryngology and Otology. Overall, there were 45 articles of two pages or less. An appropriate summary existed in 79.6% of articles published in Auris Nasus Larynx and in 33.9% of abstracts located in The Journal of Laryngology and Otology. A unique occurrence was reported in 14 and 27.82%, respectively. The number of citations ranged from 3 to 28 (mean 12.4) in Auris Nasus Larynx and 2 to 28 (mean 10.2) in The Journal of Laryngology and Otology. CONCLUSION: Case reports require a more applicable summary with a limit on authors, pages and references. PMID- 15364353 TI - Evaluation of missing fundamental phenomenon in the human auditory cortex. AB - OBJECTIVE: Harmonic complex tones consisting of four or more continuous harmonics of a certain stem frequency are perceived as the pitch of the fundamental frequency tone, it is referred to as the missing fundamental phenomenon (MFP). It is considered that the MFP is produced in the central auditory system, not in the periphery. However, it remains unclear where and how complex sounds is integrated. Using 306ch magnetoencephalography (MEG), we investigated when and where the MFP was integrated in the auditory cortex. METHOD: We examined six subjects who were selected by MEG in 12 healthy right-handed adult volunteers with normal auditory sensation. Ears were randomly stimulated with five different complex tones consist of fundamental frequency tone and harmonic complex tones. The location and direction of equivalent current dipoles (ECD) were evaluated at P50 and N100 in the right temporal lobe by MEG. Dispersion of the source of ECD was respectively evaluated on their brain MRI. RESULTS: Stimulation of ears with harmonic complex tones and the stem frequency tone revealed the localization of P50 and N100 ECD in the transverse temporal gyrus and their peripheral superior temporal gyrus. Although the sources of P50 ECD for harmonic complex tones and the fundamental tone were varied around the transverse temporal gyrus and superior temporal gyrus, the sources of N100 ECD were almost identical at the transverse temporal gyrus, demonstrating the MFP. This phenomenon were similarly observed, even when dichotic listening were stimulated. CONCLUSION: These findings suggest that the MFP occurs in the transverse temporal gyrus and the superior temporal gyrus, which are the primary auditory cortex, between P50 and N100. PMID- 15364354 TI - Vestibular-like facial nerve schwannoma. AB - OBJECTIVE: To evaluate the prevalence as well as the management of primary facial nerve schwannoma (PFNS) presenting as vestibular schwannoma. METHODS: A retrospective review was carried out on 146 patients referred to and operated upon in our institution over a 5-year period for tumors of the internal auditory canal extending or not into the cerebello-pontine angle (TICA). A review of the literature included an extensive MEDLINE search of the literature since 1966 and careful inspection of the bibliography of each article. Articles pertaining to facial nerve schwannomas were reviewed to identify cases of vestibular-like facial (V-LF) nerve schwannoma. RESULTS: Two cases of tumors were finally diagnosed as PFNS. Clinical and radiological evaluations were typical of a vestibular schwannoma. Facial function was normal in each case. Unsuspected facial nerve schwannomas were found during surgery. Tumor removal was not performed. Postoperatively House and Brackmann (HB) facial function was normal in one case and grade II in the other case. These two cases represented a 1.4% prevalence, which is within the limits reported in the literature. Among V-LF nerve schwannomas reported in the literature, the postoperative functional facial outcome usually worsened when the tumor was resected. CONCLUSION: The prevalence of V-LF nerve schwannoma has been reported in the two last decades owing to the development of modern imaging. The conservative management of early staged tumors presenting as vestibular schwannoma is recommended in order to counsel the patient regarding the possibility of V-LF nerve schwannoma and to prolong normal facial function. PMID- 15364355 TI - Eye movements evoked by selective saccular nerve stimulation in cats. AB - OBJECTIVE: Because of technical obstacles in controlling current spread to adjacent peripheral nerve, eye movements evoked by activation of the otolith organs have not been investigated in detail compared to eye movements evoked by activation of the canal organs. We attempted to solve this problem by applying more sensitive methods using fine needle and strictly controlling stimulus current intensity compare with filed potential for selective stimulation. METHODS: Eye movements evoked by selective, unilateral saccular (SAC) nerve stimulation were investigated using both electrooculography (EOG) and video recording in decerebrated cats in the presence or absence of anesthesia. Electrical stimulation was applied to the SAC nerve through implanted acupuncture needles. RESULTS: In the absence of anesthesia and with stimulus intensities less than (3.1 +/- 2.7) x N(1)T, we found supraduction in both eyes or in either the ipsilateral or contralateral eye of different cats. We observed downward eye movements using a stronger stimulus intensity ((6.2 +/- 2.9)) x N(1)T). The threshold for downward eye movements was significantly greater than that for upward eye movements (P < 0.05). In anesthetized cats, only downward eye movements were observed when stimulus intensities less than 10 x N(1)T ((7.8 +/- 2.3) x N(1)T) were used. CONCLUSION: These results confirm the known sacculo ocular anatomical connections, which are involved predominantly in vertical eye movements. Because the sacculo-ocular connections are relatively weak, the normal supraduction evoked by SAC activation can be easily modified by factors such as level of anesthesia and the method of stimulation. PMID- 15364356 TI - Efficacy of misoprostol in the treatment of tinnitus in patients with diabetes and/or hypertension. AB - OBJECTIVE: To determine the efficacy of the prostaglandin E1 analogue misoprostol in the treatment of tinnitus in diabetic and/or hypertensive patients. DESIGN: Double-blind, randomized, placebo-controlled trial. SETTINGS: Tertiary care referral center. METHODS: The subjects were 42 patients with hypertension and/or diabetes mellitus who had chronic tinnitus and had experienced tinnitus symptoms for a minimum of 6 months. Twenty-eight patients were randomly assigned to Group I (misoprostol treatment), and 14 patients to the Group II (placebo treatment). Misoprostol therapy was started at 200 microg per day, and was increased 200 microg every 7 days until a dose of 800 microg per day was reached. The same numbers of placebo tablets were given to the control group using the same schedule. Both groups were treated for 1 month. The changes in objective and subjective tinnitus findings from baseline to 1 month were assessed, and the group results were compared. The chi(2)-test, student's t-test and paired-samples t-test were used to analyze the study. RESULTS: At the completion of treatment, objective assessment showed that tinnitus loudness decreased in 13 (46%) of the 28 patients in the experimental group, whereas this was observed in only two (14%) of the 14 subjects in the placebo group. Subjective tinnitus scoring revealed improvement rates of 29 and 14% for the misoprostol and placebo groups, respectively. When t-test relating to difference between rates were performed, the difference between improvement rate for tinnitus loudness of the experimental group and control group was found to be statistically significant (P = 0.05), but difference between improvement rate based on subjective tinnitus scoring was insignificant (P = 0.22). CONCLUSION: Misoprostol is an effective and safe treatment for chronic tinnitus in hypertensive and/or diabetic patients. Our results are encouraging, but further studies of larger series are needed. PMID- 15364357 TI - Malignant melanoma of nasal cavity: report of 16 Japanese patients. AB - BACKGROUND: Primary malignant melanoma of the nasal cavity is a rare disease that has a poor prognosis. There are significant racial differences in the incidence of melanomas in the nasal cavity. METHODS: Sixteen Japanese patients treated at the Hospital of Tokyo University were reviewed retrospectively. RESULTS: Fourteen patients were treated with surgery. Two patients received treatment with radiotherapy alone. Seven patients had microscopically negative surgical margins and seven patients had positive surgical margins. The 2- and 5-year actuarial survival rates for all patients were 63.6 and 31.8%, respectively. No statistically significant difference could be shown in the overall survival rate between patients with positive surgical margins and those with negative surgical margins. CONCLUSIONS: Clinical features of nasal melanoma in Japanese are not different from nasal melanoma in Caucasians. The negative surgical margins were not predictive of a better prognosis. PMID- 15364358 TI - Antitumor and antiangiogenic effects of interleukin 12 gene therapy in murine head and neck carcinoma model. AB - Interleukin-12 (IL-12) plays a critical role in producing an immune response, as indicated in many ways, e.g., induction of interferon-gamma (IFN-gamma), and augmentation of the cytotoxic activity of resting activated T cells and natural killer (NK) cells. In this study, we examined whether intratumoral injection of a recombinant retrovirus vector expressing IL-12s induce antitumor and antiangiogenic effects in a murine model using a murine head and neck squamous cell carcinoma (NR-S1). In vitro the levels of vascular endothelial growth factor (VEGF) mRNA and protein expression were decreased in IL-12 gene transfected NR-S1 cell. in vivo direct IL-12 gene therapy resulted in significantly remarkable inhibition of tumor growth compared to the control group. The tumor regression by direct IL-12 gene therapy was also associated with decreased vessel density, and apoptosis and increased infiltration of CD8(+) T cells and CD56(+) NK cells in the tumor increased. Also, the number of IFN-gamma expressed cells of spleen cells was increased in the treatment group compared with the control group. These results suggested that direct IL-12 gene therapy appears to be effective in reducing tumor growth by triggering both antiangiogenic effects and an immunological enhancing mechanism through induction of IFN-gamma. PMID- 15364359 TI - Longitudinal analysis of progression of dysphagia in amyotrophic lateral sclerosis. AB - OBJECTIVE: To investigate the progression of dysphagia in amyotrophic lateral sclerosis (ALS) using videofluoroscopy (VF), according to two different scales: the duration following bulbar symptom onset, and the ALS severity scale (ALSSS). METHODS: Seventy-two VF examinations, on 50 ALS patients, were performed at various time points, and the change in VF findings in the oral and pharyngeal phases was analyzed over time following bulbar symptom onset. The change according to advancement of the ALS severity scale was also assessed. RESULTS: Delayed bolus transport from the oral cavity to the pharynx, and bolus stasis at the pyriform sinus (PS), were seen in about half of the patients with no bulbar complaints. In contrast, upper esophageal sphincter (UES) opening was relatively well maintained in the late stage of dysphagia. The other parameters, such as bolus holding in the oral cavity, constriction of the pharynx, and elevation of the larynx, became worse over time following bulbar symptom onset. These parameters on VF examination also became worse as the ALSSS advanced; however, even patients in the group with normal eating habits showed disturbed bolus transport from the mouth to the pharynx, weak constriction of the pharynx, and bolus stasis at the PS. Generally, dysfunction had progressed dramatically, in most of the parameters, in the patients that required modification of food; however, UES opening and elevation of the larynx were relatively well maintained in advanced-ALSSS patients. CONCLUSION: In this study, we clearly show the progression pattern of dysphagia in ALS, and we also emphasize that VF is a useful tool to follow swallowing function in ALS patients. From our results, it would be useful to perform VF by 6 months following bulbar symptom onset, because VF can contribute to rehabilitation or appropriate diet modification; and later, around 1 year following bulbar symptom onset, VF should again be performed, since ALS patients show wide-ranging nutrition intake patterns, and they need assessment to introduce intervention, such as tube feeding, according to their varying status of dysphagia, around at this time point. PMID- 15364360 TI - Observation of tongue papillae by video microscopy and contact endoscopy to investigate their correlation with taste function. AB - OBJECTIVES: Contact endoscopy is a technique used to obtain detailed images of living epithelium in the fields of gynecology, rhinology and laryngology. Video microscopy is useful for observation of the surface of tongue papillae. In the present study, we attempted to apply the contact endoscopy technique together with video microscopy to observe tongue papillae, and to study the correlation between the condition of fungiform papillae and taste function. METHODS: Ten subjects (3 men, 7 women) were divided into two groups based on the results of a taste examination by electrogustometry. We compared the shape and blood vessels of the papillae between normal taste and taste disorder groups. RESULTS: In the normal taste group, round shaped papillae and clear blood vessels were observed with both microscopy and contact endoscopy. In the taste disorder group, flat and irregular papillae were observed with microscopy. Blood vessel flow of the papillae was observed to be poor with contact endoscopy. CONCLUSION: These findings suggested that the images of microscopy and contact endoscopy were related to taste function, and both techniques were useful for evaluating taste function. PMID- 15364361 TI - Identification of differentially expressed genes in salivary gland tumors with cDNA microarray. AB - OBJECTIVE: The final goal of this study is to develop a pre-operative fine needle aspiration biopsy (FNA) diagnostic system based on gene expression profiles. As the first step to that end, the present study was performed to determine whether the cDNA microarray system is applicable for histological evaluation of parotid gland tumors. METHODS: We investigated molecular characteristics on the basis of gene expression patterns of the two most common types of salivary gland tumors (pleomorphic adenomas and Warthin tumors) and normal salivary gland tissues, using the cDNA microarray system. RESULTS: Pleomorphic adenomas and Warthin tumors can be classified by cDNA microarray. In pleomorphic adenomas, 11 independent genes were found to be up-regulated and 2 genes were down-regulated. In Warthin tumors, five independent genes were found to be up-regulated, and six genes were down-regulated. In hierarchical clustering analysis, cases were further grouped into two clusters according to the histological type. Furthermore, cDNA microarray enabled pleomorphic adenomas to be subclassified into three clusters according to the histological subtypes. CONCLUSIONS: This study suggested that cDNA microarray may be useful and applicable for the pre operative diagnosis (such as FNA) of the salivary gland tumor. PMID- 15364362 TI - Use of free flaps for reconstruction in head and neck surgery: a retrospective study of 182 cases. AB - OBJECTIVE: In order to clarify the factors affecting the survival of flaps, we performed a retrospective study on free flaps (187) used for reconstruction in head and neck surgery in 182 patients. METHODS: Free jejunum flaps were used on 68 occasions, rectus abdominus myocutaneous flaps on 67, radial forearm flaps on 49, scapular osteocutaneous flaps on 2 and latissimus dorsi myocutaneous flap on ones occasion, during the period from May 1996 to April 2003. Post-operative circulatory complications at the recipient site were analyzed mainly in relation to a history of previous surgery, radiotherapy and chemotherapy. RESULTS: Circulatory crisis was observed in seven cases and circulation was restored in two of them after emergency exploration, whereas the other five flaps failed to survive. The overall failure rate of free flaps was 2.7%. Wound infection at the recipient site was frequent in patients with a history of previous local cervical surgery. Flap failure was significantly more frequent in patients with a history of previous surgery and infection, except for 2 patients in whose case the microsurgical technique was inadequate. Flap failure was not observed in those patients whose irradiation field could be analyzed, or in the patients who received chemotherapy. Although the incidence of circulatory crisis as a consequence of inadequate microsurgical technique was observed in the early period of this retrospective study, their frequency has diminished in the last 2 years. CONCLUSION: The survival of flaps was affected by a history of previous surgery following wound infection, although a history of irradiation and chemotherapy would have no effects on flap failure. The appropriate choice of vessels and surgical skill are crucial for the success of free flaps from our experience. Emergency exploration was also crucial for the survival of the flap in the case of circulatory crisis. PMID- 15364363 TI - Intractable epistaxis related to cavernous carotid artery pseudoaneurysm: treatment of a case with covered stent. AB - Although epistaxis is commonly seen in otorhinolaryngologic practice, it may rarely become a severe and life-threatening problem. Pseudoaneurysm of internal carotid artery (PICA) is an uncommon, but potentially fatal cause of epistaxis. Optimal management demands rapid recognition and treatment to give the best functional outcome. High suspicion and carotid arteriography is essential for diagnosis. In this article, a 28-year-old male case with post-traumatic cavernous PICA localized to sphenoid sinus and presenting with intractable epistaxis was reported. The patient was managed by endovascular covered stent application successfully. In differential diagnosis of patients with intractable epistaxis and isolated sphenoid sinus lesions, PICA should be considered. Early treatment with covered stent of aneurysm can be a life saving therapeutic approach. PMID- 15364364 TI - Rosai-Dorfman disease revealed in the upper airway: a case report and review of the literature. AB - Rosai-Dorfman disease (RDD) is a systemic disease and its etiology is not well understood. It is a very rare but established disease. We report a case of RDD diagnosed in the upper airway that became a life-threatening condition because of the stricture of the subglottic space. A 49-year-old female complained of nasal obstruction, hoarseness and lacrimation. After evaluation following by fiberscopic examination and CT scanning of the head and neck, sinusitis and laryngeal masses were detected. Endoscopic sinus surgery and endolaryngeal microsurgery were performed simultaneously. The masses in the nasal cavity and larynx showed the same histological findings. Proliferative large histiocytes with voluminous clear cytoplasm and rounded nuclei, and lymphophagocytosis known as emperipolesis are the characteristic features of RDD. In this case, extranodal manifestations in the subglottic space gradually emerged postoperatively. Tracheotomy was performed to ensure an airway and prevent suffocation. An extensive treatment is not required in most patients with RDD, however, some patients who have extranodal manifestations involving the airway may detoriorate into a life-threatening condition. PMID- 15364365 TI - Well-circumscribed intramuscular lipoma of the sternocleidomastoid muscle. AB - Intramuscular lipomas are unusual benign mesenchymal tumours, which infiltrate the skeletal muscle, and are exceedingly rare in the head and neck region. They commonly infiltrate the skeletal muscle fibres from which they arise and are rarely well circumscribed. We present the only documented case of well circumscribed intramuscular lipoma arising from the sternocleidomastoid muscle. Although the recurrence is commoner in the infiltrative variety the surgeon should be aware that differentiation between infiltrative and well circumscribed is based on histological diagnosis, and hence wide excision in all cases of intramuscular lipomas is essential. PMID- 15364366 TI - Thyroid anaplastic carcinoma transformed from papillary carcinoma in extrathyroid area. AB - We report a 75-year-old male with anaplastic carcinoma in an extrathyroid area. Thyroid remained unchanged. The patient is alive without incident of tumor recurrence at 3.5 years after total resection and at 5 years after initial symptom. The tumor developed between the sternocleidomastoid muscle and common carotid artery, and was completely separated from the thyroid. The tumor location was consistent with a branchial cyst. The tumor consisted of two parts; an upper solid tumor and a deep cystic tumor. The former showed anaplastic carcinoma with osteoclast-like giant cells. The latter was consistent with thyroid papillary carcinoma. The center was intermingled with these two carcinomas. Anaplastic carcinoma cells were positive for vimentin and papillary carcinoma cells were positive for keratin, thyroglobulin, and thyroid transcription factor-1. These results remain insufficient to find any conclusions concerning the tumor nature; either ectopic thyroid carcinoma arising from a branchial cyst or occult thyroid carcinoma metastasis. This is rare case in which thyroid anaplastic carcinoma transformed from papillary carcinoma in an extrathyroid area. PMID- 15364367 TI - Warthin's tumor of the nasopharynx: a case report. AB - Warthin's tumor in the nasopharynx is extremely rare. A 76-year-old man was referred to our hospital for treatment of a nasopharyngeal tumor. Fiberscopic examination, CT and MRI findings showed an oval mass in the right side of the posterior nasopharynx. The tumor was resected surgically using the transpalatal approach. Pathological examination indicated Warthin's tumor. There have not been any signs of recurrence postoperatively. PMID- 15364368 TI - Normocalcemic hyperparathyroidism presented with mandibular brown tumor: report of a case. AB - Brown tumor is a rare clinical entity complicating hyperparathyroidism. It may occur in the head and neck, with the mandible being the most frequent site. Hyperparathyroidism is usually associated with hypercalcemia. We report a case of madibular Brown tumor secondary to primary hyperparathyroidism. In this case in spite of hyperparathyroidism and the bony lesion the serum calcium level was within normal range. The case managed by surgical excision of the mandibular tumor with an en-bloc hemithyroidectomy with inclusion of the diseased parathyroid gland. This case demonstrates that in osteolytic bony lesions a hyperparathyroid complication can be expected even with normal serum calcium level. The presence of normocalcemia in primary hyperparathyroidism should prompt the physician to look for vitamin D deficiency. PMID- 15364369 TI - Extravascular papillary endothelial hyperplasia arising from parapharyngeal space. AB - A 66-year-old man visited our clinic suffering from swelling of right pharyngeal space. He was not aware of other symptoms. The contrast CT showed patchy enhancement with calcification. The enhanced MRI revealed a non-homogeneous signal pattern and patchy central enhancement. During surgery, egg-shaped hard elastic tumor was observed in parapharyngeal space without any sign indicating an invasion into surrounding nerves or feeding vessels. Then the tumor was removed without significant hemorrhage or damage to the cranial nerve. Histological examination of the tumor revealed a nodular lesion with blood clot and hematoma encapsulated with fibrous tissue having vascular spaces of irregular shape and size. From those findings, the tumor was diagnosed as papillary endothelial hyperplasia (PEH). However, since the location of the tumor differed from that of PEH commonly observed in the intravascular space, the tumor was classified as extravascular PEH. PMID- 15364370 TI - A case of Kuttner tumor of the submandibular gland. AB - Kuttner tumor (known as chronic sclerosing sialadenitis) is a totally benign inflammatory lesion and to date there has been no report of malignancy. However, because of its clinical features, the clinical diagnosis is often that of a salivary gland neoplasm. We report a case of Kuttner tumor and discuss its differential diagnosis, especially the pathological diagnosis. Kuttner tumor may be mainly due to T lymphocyte immune reaction and of itself is a benign lesion, but it may provide a state in which a malignant lesion arises. PMID- 15364371 TI - Two cases of pulmonary embolism after head and neck surgery. AB - Pulmonary embolism (PE) is a prominent cause of morbidity and mortality in surgical patients. Here, we report two cases of PE following head and neck surgery: (1) in case 1, the patient underwent endoscopic sinus surgery (ESS), and (2) in case 2, the patient was suspected PE after resection of the parotid gland and radical neck dissection. Prophylactic compression devices were used during the operation in both cases. In case 1, PE was diagnosed 3 days following surgery after the patient complained of dyspnea while walking. PE was successfully treated by intravenous administration of heparin. In case 2, PE was strongly suspected 13 days following surgery after the patient experienced syncope and chest pain and exhibited abnormal findings on a cardiac echogram. This patient died 18 h after the onset of cardiac symptoms. Despite prophylactic measures taken during surgery, the prognosis was poor for this patient due to numerous high-risk factors (e.g., advanced age, obesity, prolonged immobilization). Although the incidence of postoperative PE in our department during the last 7 years is very low (0.04%), the frequency of postoperative PE in Japan has steadily increased in recent years. PE still is one of the most significant complications leading to morbidity and mortality following surgery. Given the gaining prominence of PE, we conclude that otolaryngologists including those in Japan should be reminded of high-risk factors associated with PE and be made aware of prophylactic treatments newly aimed at reducing the frequency of PE. PMID- 15364372 TI - Intramuscular myxoma of scalene muscle: a case report. AB - Intramuscular myxoma is a benign mesenchymal tumor that commonly occurs in the larger skeletal muscles, and is rare in the head and neck. A case of intramuscular myxoma in the scalene muscle is described. This is the first report of this tumor arising in that muscle. It is difficult to diagnose pre-operatively from clinical features, CT scan and ultrasonography of the neck because of the lack of characteristic manifestation or radiographic features. Intramuscular myxoma has an infiltrative tendency, so there are several reports of recurrence because of incomplete excision or enucleation. In our case no recurrence was observed for 4 years, since the tumor was removed with the muscle adherent to it. PMID- 15364373 TI - Postauricular subcutaneous leiomyosarcoma. AB - Superficial leiomyosarcomas are rare soft tissue tumors, most frequently involving the lower extremity. Leiomyosarcomas which could be misdiagnosis or delayed are very rare in the head and neck region. In this report, a 46-year-old man with postauricular leiomyosarcoma was presented. The tumor was totally excised with 3 cm margins. Immunohistochemically, the specimen showed reactivity for smooth muscle actin, vimentin, and desmin. No complications developed in the postoperative period and the patient was free of recurrence at the 26 months follow-up. Immunohistopathological examination is essential for an accurate histological diagnosis. The ENT surgeon should pay attention to superficial leiomyosarcoma with its specific behavior and treatment while evaluating the lesions in the head and neck region. PMID- 15364377 TI - On the prehistory of the German Society of Legal Medicine. AB - The paper gives a review on the foundation of local associations of legal medicine and their journals at the beginning and in the mid of the 19th century in Germany before the German Society of Legal Medicine was founded. PMID- 15364378 TI - The history of the German Society of Forensic Medicine. PMID- 15364379 TI - Portraits of some founders of the German Society of Legal Medicine. AB - In the preceding paper by W. Krauland the foundation of the German Society of Legal Medicine, which took place at the 76th meeting of the German Naturalists and Physicians in Breslau, 1904, was described in detail. Carl Ipsen, Julius Kratter, Adolf Lesser, Georg Puppe, Emil Ungar and Fritz Strassmann were chosen as the preliminary board of the Society. They were respected scientists in their field, working at different universities in Germany and Austria. Life and scientific work of these founders of the German Society of Legal Medicine shall be described in short portraits. PMID- 15364380 TI - The history of forensic medicine in times of the Weimar republic and national socialism--an approach. AB - German Forensic Medicine established the tasks and aims of forensic medicine during the period of the "Weimar Republic" (1919-1933). Since 1925, forensic medicine is part of the medical examination regulation. In times of the 3rd Reich (1933-1945), most of forensic pathologists were influenced by and involved in National Socialism. The special subject of forensic medicine was endangered by the absence of political neutral forensic pathologists. PMID- 15364381 TI - Persecution of Jewish forensic pathologists. AB - Not even forensic pathologists were spared by the anti-Jewish laws of the Third Reich. Fritz Strassmann and Paul Fraenckel were among more than 140 faculty of the Berlin Department of Medicine persecuted by the national socialist dictatorship. It was because of their Jewish background that Georg Strassmann was expelled from university in Breslau, and Leone Lattes was forced to leave in Pavia. Miklos Nyiszli was deported from Oradea to Auschwitz and forced to perform forensic autopsies. Stefan Jellinek in Vienna, Ludwik Hirszfeld in Warsaw and Friedrich Schiff in Berlin were other medical professionals whose achievements had enriched legal medicine before they became victims of anti-Jewish persecution. PMID- 15364382 TI - Forensic medicine in the German Democratic Republic. AB - Forensic medicine services were resumed in the territory of the Soviet occupation zone in early summer 1945. One coherent legal system was established, following the foundation of the German Democratic Republic (GDR), 7 October 1949. Forensic medicine was subject of teaching and examinations from the very outset. Sub specialisation (fellowship) was introduced in 1955. The Gesellschaft fur Gerichtliche Medizin der DDR (GDR Society of Forensic Medicine) was established in 1967 and began to organise with high continuity a wide-ranging variety of scientific events. Almost all administrative districts had their own institute of forensic medicine by 1990. PMID- 15364383 TI - Legal medicine in the Federal Republic of Germany and after reunification. PMID- 15364384 TI - Austrian forensic medicine. PMID- 15364385 TI - Legal medicine in Switzerland. PMID- 15364386 TI - Research in forensic neurotraumatology. AB - Over the past 100 years forensic research in neurotraumatology was focusing on the genesis, e.g. biomechanis, and the origin of epidural, subdural, subarachnoidal, intracerebral and brain stem haemorrhage, particularly under aspects to enable the differential diagnosis of bleeding due to non-traumatic diseases. Moreover the estimation of the age of brain injuries has important criminological implications (survival time following traumatic forces to the head, alibi etc.). Beside these main fields of research, aspects of expertise in special areas such as head trauma due to child abuse, capability to act despite severe brain injuries and research on cervical trauma are reviewed. PMID- 15364387 TI - Estimation of the time since death in the early post-mortem period. PMID- 15364388 TI - Blood alcohol/congeners of alcoholic beverages. PMID- 15364389 TI - History of forensic serology and molecular genetics in the sphere of activity of the German Society for Forensic Medicine. AB - In the field of forensic serology, essential developmental impulses have come from the sphere of activity of the German Society for Forensic Medicine. Among these are the orientating enzyme-reactive and specific tests for blood using crystallization tests, the determination of the species-specificity of the donor of the stain and the beginning of the individualization of a stain to its donor. Pioneering work has also been done in the development of blood group serology of the conventional markers. DNA analysis originated in the Anglo-Saxon region. The German Society for Forensic Medicine also contributed to its further progress and essentially influenced it e.g. by the discovery of microsatellite markers, including STRs, by the validation of numerous test methods, by optimization of these methods and by preliminary work for the use of mass spectrometry in DNA analysis. PMID- 15364390 TI - Traumatology of the traffic accident--dead people for the safety in traffic. AB - Since 1970, traumatomechanics has been a focal point in research at the Institute for Legal Medicine and Traffic Medicine in Heidelberg. Here, the main topics are the understanding of the interrelation between mechanical strain and the resulting degree of injury; at the forefront of all interest is the determination of the mechanical resilience in humans, their organs and tissues. Important are not only the means, but likewise the individual strain tolerance and the causes for its variability. Their understanding leads to scientifically justifiable expert's reports. In safety research, these data are of major importance for the validation of crash-dummies and for the improvement of safety protection systems. Before this background, national and international institutions have supported numerous projects. With the help of 2 examples, the Thorax-Trauma-Index (TTI) and the synergy between safety belt and airbag, the relevance of these data for international regulatory provisions and the progress in safety practice are illustrated. Some traumatomechanical insights can only be gathered from human corpses. Legal prerequisites and ethical problems of experiments with corpses are discussed. PMID- 15364391 TI - Gunshot injuries as a topic of medicolegal research in the German-speaking countries from the beginning of the 20th century up to the present time. AB - The 100th anniversary of the foundation of the German Society of Legal Medicine is a good opportunity to review its contributions to forensic wound ballistics. The present article gives an overview of the scientific development in this field with special emphasis on work pioneering new developments and findings valid up to the present day, for example the presence of carboxyhemoglobin in the vicinity of the entrance wound as a sign of a contact or close-range shot [A. Paltauf, Wien. Klin. Wochenschr. 3 (1890) 984-991, 1015-1017]; the correct interpretation of the muzzle imprint [A. Werkgartner, Beitr. Gerichtl. Med. 6 (1924) 148-161] and the retrograde ballooning of the bullet entrance region in contact shots [F. Hausbrandt, Dtsch. Z. ges. Gerichtl. Med. 38 (1944) 45-76; H. Elbel, Med. Welt 20 (1958) 343-345]; wound patterns from captive-bolt livestock stunners [H. Czursiedel, Dtsch. Z. ges. Gerichtl. Med. 28 (1937) 132-133]; singeing of synthetic fiber textiles in close-range shots with nitro powder ammunition [S. Berg, Arch. Kriminol. 124 (1959) 5-8,17-22]; the wound ballistic processes on penetration of the bullet and the origin of the abrasion collar [K. Sellier, Beitr. Gerichtl. Med. 25 (1969) 265-270]. More recently medicolegal research in the German-speaking countries covered the following subjects: studies of the dynamic bullet-target interactions in experimental gunshots to simulants and composite body models; use of modern imaging techniques (CT, MRI) in the pre autopsy diagnosis of lethal gunshot injuries; injuries from blank guns; mechanisms of incapacitation by gunshot injuries; development of improved methods for the evidence of gunshot residues on the firing hand; backspatter from close range shots; medicolegal contributions in the discrimination of accidental, homicidal and suicidal gunshot injuries. PMID- 15364392 TI - Asphyxia-related deaths. PMID- 15364393 TI - 100 Years of toxicology in the German Society of Legal Medicine. PMID- 15364394 TI - Vitality and time course of wounds. AB - The term "wound" describes the morphologic-functional disruption of the continuity of a tissue structure. A wound can be inflicted during life--when the cardiovascular and respiratory system is still intact--or after death, i.e. after cardiac and respiratory arrest. Traumatization during life triggers vital reactions that do not occur in postmortem wounds. Three types of vital reactions in wound healing can be distinguished: Reactions of the scavenger type, which are almost exclusively mediated by blood cells. Reactions by complex signal transduction pathways, which involves cascade-like release of chemokines, cytokines and adhesion molecules and may influence type 1 and type 3 reactions. Reactions of the scarring type, which involve the final repair of the damaged tissue and are carried out primarily by cells residing at the wound edges, i.e. partly concerning mesenchymal cells and partly tissue-specific cells dependent on the involved organ system. The three different types of reaction follow roughly parallel temporal courses that include cascade-like interactions among themselves. Whereas demonstration of a vital reaction suffices to differentiate an intravital wound from a postmortem wound, the vital reactions themselves follow strictly temporal courses. The regular time-dependent occurrence of each phenomenon allows--in limits--a reliable temporal classification of wound healing. A review will be given especially demonstrating the actual German scientific research in vitality and in skin wound timing as well as in timing of mechanical injury of the brain. PMID- 15364395 TI - Forensic radiology in German-speaking area. PMID- 15364396 TI - Forensic age diagnostics of living people undergoing criminal proceedings. AB - In the German-speaking area, recent years have seen a rapid growth of the need for forensic age estimations. Such need arises, for example, if no verified information on the age of a person suspected of a criminal offence is available; the issue at question in terms of criminal law is whether the person concerned has reached the age of criminal responsibility and whether general criminal law in force for adults is to be applied. The relevant age tresholds in criminal proceedings are 14, 18 and 21 years of age. According to recommendations of the Study Group on Forensic Age Diagnostics, a forensic age estimate should consist of a physical examination, an X-ray of the hand, and a dental examination which records dentition status and evaluates an orthopantomogram. In addition, a radiological or CT examination of the clavicles is recommended to establish whether a person has attained 21 years of age. The present article addresses the influence of ethnic origin on the examined developmental systems. PMID- 15364397 TI - Sudden infant death--focus subject of medico-legal research. AB - The compilation of research activities concerning SID in the field of legal medicine could only choose those out of a great abundance of high-quality examinations which represent applied research. By this they have contributed either to the clarifying of the aetiology which has not been cleared up or to the prophylaxis. For this looking through in a widest sense the basic approach was to consider SID being the result of an intrinsic and/or extrinsic disturbance of the modulation of respiratory regulation of the infant. This namely means a metabolic disturbance of respiratory neurons. One must not share this opinion even if it is given a certain plausibility by newer physiological examinations. But this also means that some examinations did not receive the acknowledgement which they would have received if there had been an immunological approach. The compilation was completely done without a presentation of the primary crisis intervention and the long-term care as being a fundamental medico-legal approach. Altogether may be summarized that legal medicine has completely fulfilled its duty to take care of the problem SID and that the examinations did not remain without success. PMID- 15364398 TI - The contribution of forensic medicine to forensic anthropology in German-speaking countries. PMID- 15364399 TI - The history of forensic entomology in German-speaking countries. AB - Megnin's book "La fauna des cadaveres" published in 1894 in France is generally accepted as a mile-stone in forensic entomology. It is hardly known that at the same time this topic was likewise explored in the German-speaking countries. Even PMI estimation based on developmental data of blowflies was performed. After a more descriptive period in the first half of the 20th century the complexity and variability of insects' biological behavior were detected and formally investigated. Improved technical facilities, enhanced comprehension of scientific studies and multidisciplinary cooperation, enabled rapid progress in forensic entomology during the last decades. With the European Association for Forensic Entomology founded in 2002 the frame work for a high standard of competency at an international level was constituted. PMID- 15364400 TI - Malpractice--role of the forensic pathologist in Germany. AB - Medicolegal malpractice assessments have been an important part of the work of forensic pathologists. Not only botchers are concerned but also physicians of all clinical subjects, especially malpractice charges claiming a malpractice leading to death. Single long articles and book chapters have been published about malpractice assessments since the end of the 17th century by forensic pathologists. After World War II systematic studies of malpractice charges first were published in the 1960s. Meanwhile the attention focuses more and more on the role of forensic pathologists to help in prevention of malpractice. PMID- 15364401 TI - Clinical forensic medicine and its main fields of activity from the foundation of the German Society of Legal Medicine until today. AB - The fields mainly covered by clinical forensic medicine are subject to time related changes which are described on the basis of the German literature of the 20th century. Some fields of forensic sexual medicine (diagnosis of virginity, proof of criminal abortion, potentia coeundi, potentia generandi, potentia concipiendi) have become less important in the daily work of medicolegal institutes, whereas victims of rape and sexual abuse continue to form a major part of the forensic examination material in the German-speaking countries. The evaluation of suspected physical child abuse has grown in importance since the 60s, and it is essentially the merit of Elisabeth Trube-Becker that this problem is now dealt with also in scientific medicine. More recently, medicolegal experts are increasingly confronted with further groups of persons: victims of domestic violence, abused/neglected seniors, refugees from countries where torture is used. A new special field, which established itself only in the 90s, is the estimation of age with regard to the criminal responsibility of suspects who have no identity papers or pretend to have none. A phenomenon frequently observed in the last two decades is the non-accidental self-infliction of injuries. Whereas in the first half of the 20th century the motive for self-mutilation was typically to evade military service, this category of injury was later mainly seen in the context of insurance fraud; more recently most forensically relevant self-inflicted injuries refer to simulated offenses (fictitious sexual offenses and robbery, attacks allegedly having a political background). One of the traditional fields of clinical forensic medicine continues to be the evaluation of victims and suspects following bodily harm and attempted homicides. In the field of civil law medicolegal experts are particularly often concerned with controversial consequences of traffic accidents (e.g. alleged whiplash injuries after rear-end collisions at low velocities). PMID- 15364402 TI - The centenary of the German Association of Legal Medicine--from its foundation in 1904 to 21st century Legal Medicine. PMID- 15364403 TI - Textbooks on legal medicine in the German-speaking countries. AB - In this survey, the textbooks on legal medicine which were published in the German-speaking countries from the 17th century up to now are briefly reviewed. The textbooks with the most impact on the further development of legal medicines were those published by Johann Ludwig Casper in Berlin and Eduard Ritter von Hofmann in Vienna in the 19th century. The main textbooks of the 20th century are addressed as well, also the different development in both German countries. PMID- 15364404 TI - Influence of maternal pineal gland on the developmental pattern of neurokinin A (NKA) and substance P (SP) in male-rat-offspring: relationship to the season of the year. AB - The present study examines the influence of maternal pineal gland on the frontal cortex, striatal and testicular concentrations of the tachykinins, neurokinin A (NKA) and substance P (SP). Control, pinealectomized (PIN-X) and PIN-X plus melatonin-treated (PIN-X + MEL) mother rats were prepared. Male offspring rats were studied at 21, 31 and 60 days of age, during the four seasons of the year. In control-offspring tachykinin concentrations in frontal cortex were found at their highest levels in 21-day-old rats with a moderate decrease up to 60 days of age. This developmental pattern was season-dependent, observed only during summer and fall. Maternal PIN-X or PIN-X + MEL resulted in alterations in the offspring, showing during spring and summer significantly higher concentrations (P < 0.01) and during fall significantly lower concentrations of tachykinins in the frontal cortex (P < 0.05, P < 0.01) as compared to control-offspring. The tachykinin concentration in the striatum of control-offspring showed no major modifications throughout the ages studied in the four seasons of the year. With very few exceptions, PIN-X- and PIN-X + MEL did not alter tachykinin concentrations in striatum. Testicular SP concentrations showed a decrease from 21 to 60 days of age. PIN-X or PIN-X + MEL only caused minor and inconsistent modifications in testicular SP levels. In conclusion, our data clearly indicate for the first time that the maternal pineal gland participates in the regulation of the postnatal tachykinin development in some areas of the central nervous system. This effect was more evident in the frontal cortex than in the striatum and testes. PMID- 15364405 TI - Expression of ether a go-go potassium channels in human gliomas. AB - Ether a go-go (EAG) K(+) channels have been shown to be involved in tumor generation and malignant growth. Gliomas have not been investigated thus far. Using RT-PCR we investigated healthy human brain and human gliomas of different subtypes and malignancy grades for the expression of human EAG1 and eag-related gene (ERG) 1 channels. mRNA of both channels was detected in all tissues. Expression was strong in normal brain, moderate in high-grade and high in low grade gliomas. Our findings suggest a differential expression of hEAG1 and hERG1 in gliomas depending on the malignancy grade and nature of the tumor cells. However, the hypothesis that EAG channels are related to the oncogenic process itself is only partly supported by this study. PMID- 15364406 TI - Long-term effects of neonatal seizures on subsequent N-methyl-D-aspartate receptor-1 and gamma-aminobutyric acid receptor A-alpha 1 receptor expression in hippocampus of the Wistar rat. AB - To evaluate the pathophysiological mechanism of subsequent reduced seizure threshold following neonatal seizures, single or recurrent prolonged seizures were induced to neonatal rats by the inhalant flurothyl. The expression of N methyl-d-aspartate receptor 1 (NR1) and gamma-aminobutyric acid receptor A-alpha 1 (GABA-A-alpha 1) immunoreactivity in hippocampus were examined by Western blotting analysis at the day 7 (P7) and day 75 (P75) after the last seizure. Whereas there were no significant changes in single seizure group and recurrent seizure group of P75, NR1 expression enhanced significantly in P7 rats of recurrent seizure group. Meanwhile, polypeptide levels of GABA-A-alpha 1 receptor subunit decreased significantly in both single and recurrent seizure-treated P7 and P75 rats. Our results suggest that recurrent or single prolonged seizures during the neonatal period may have long-term effects on the balance between excitatory NMDA system and inhibitory GABA system in hippocampus of rats. PMID- 15364407 TI - Alternative therapy of Alzheimer's disease via supplementation with choline acetyltransferase. AB - Much evidence indicates that the memory and cognitive deficits of patients with Alzheimer's disease are closely associated with dysfunction of central cholinergic system. The degree of reduction of choline acetyltransferase activity in cerebral cholinergic neurons is significantly correlated with the severity of dementia or cognitive impairments observed in Alzheimer's disease. Therefore, Alzheimer's disease may be slowed by supplementation of exogenous choline acetyltransferase. Here we show that choline acetyltransferase mediated by TAT protein transduction domain passes through the blood-brain barrier and enters the neurons in mice, increasing choline acetyltransferase and neurotransmitter acetylcholine contents. The recombination TAT-choline acetyltransferase fusion protein injected intravenously improves the memory and cognitive dysfunction in Alzheimer's disease model mice induced by amyloid-beta peptide. Our results imply a novel and potentially effective way for Alzheimer's disease therapy. PMID- 15364408 TI - Inhibition of neuropathic pain by a potent disintegrin--triflavin. AB - Injury to peripheral nerves may result in severe and intractable neuropathic pain. Many efforts have been focused on the elucidation of the mechanisms of neuropathic pain. It was found here that integrin plays an important role in the induction of neuropathic pain and treatment of disintegrin is able to attenuate neuropathic pain. The rats were induced hyperalgesia by tightly ligating the L5 spinal nerve and cut just distal to the ligature on one side. Mechanical and thermal stimuli were applied in the middle dermatome of the hind paw. Epidural administration of triflavin (TFV), an arginine-glycine-aspartic acid (RGD) containing disintegrin, inhibited hyperalgesia induced by either mechanical or thermal stimulation. Immunohistochemistry showed that the sprouting of sympathetic nerves into DRG by neuropathic surgery was markedly inhibited by TFV. Beta 1 integrin mRNA of L5 DRG increased immediately 1 day after tight ligation and cut of L5 spinal nerve. However, beta 1 integrin mRNA in uninjured L4 DRG increased later on Day 3 after surgery. On the other hand, alpha-CGRP precursor mRNA decreased in ipsilateral L5 DRG but increased in L4 DRG after neuropathic surgery. Immunohistochemistry shows that beta 3 integrins of L5 as well as L4 increased in response to neuropathic surgery and administration of triflavin antagonized the increasing action. These results suggest that there is interaction between injured and uninjured neurons and the induction of neuropathic pain is related to neuronal sprouting. Disintegrin is able to inhibit neuronal sprouting and the induction of hyperalgesia induced by peripheral nerve injury and may thus be a new category of drugs to be developed for the treatment of neuropathic pain. PMID- 15364409 TI - DRD4 exon 3 variants are not associated with symptomatology of major psychoses in a German population. AB - We previously reported an association of DRD4 exon 3 long alleles with delusional symptomatology, independently from psychiatric diagnoses [Am. J. Med. Genet. 105 (2001) 283; Psychiatry Res. 80 (1998) 129]. The aim of this investigation was to replicate these results in an independent sample from Germany. We studied 394 subjects, affected by bipolar disorder (n = 32), schizoaffective disorder (n = 45), and schizophrenia (n = 317). All affected subjects were evaluated using the Operational Criteria for Psychotic Illness (OPCRIT) checklist. DRD4 variants were not associated with symptomatology of major psychosis. Our present results, obtained in an independent German sample, did not confirm the association between DRD4 variants and delusional symptomatology. However it should be considered that the original sample included a much higher rate of mood disorders and this could partially explain the discrepancy. PMID- 15364410 TI - Increased presence of matrix metalloproteinases 2 and 9 in short- and long-term experimental herpes simplex virus encephalitis. AB - Herpes simplex virus encephalitis (HSVE) causes elevated morbidity and mortality despite antiviral treatment. Virus-independent mechanisms may perpetuate brain damage. Matrix metalloproteinases (MMPs) target extracellular matrix components. This study describes the protein and mRNA expression of MMP2 and MMP9 in experimental HSVE in the short and long term. Ten SJL-NBOM mice were infected with neurovirulent HSV-1 and compared with nine controls. The presence of MMP2 and MMP9 in brain tissue was analyzed with sodium-dodecyl-sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gelatin zymography and mRNA expression of MMP2 and MMP9 with quantitative real-time PCR at days 7, 21 and 180 post-inoculation. Infected animals had a significantly elevated gelatinolytic activity of MMP2 at all time points, and of MMP9 at days 21 and 180. Increased presence of MMP2 and MMP9 in chronic HSVE may contribute to ongoing damage. Inhibition of MMP2 and MMP9 might be a suitable target for therapeutic intervention. PMID- 15364411 TI - Aromatase, the enzyme responsible for estrogen biosynthesis, is expressed by human and rat glioblastomas. AB - The biosynthesis of estradiol and related estrogens is catalyzed by the enzyme aromatase. Among other tissues, aromatase is expressed in the brain, where it is involved in the regulation of neuroendocrine events and reproduction. Under physiological conditions, the expression of aromatase in the mammalian brain is restricted to neurons. However, recent studies have shown that reactive astrocytes express aromatase after brain injury. This opens the possibility for the expression of the enzyme in other altered forms of glial cell, such as gliomas. In the present study, the expression of aromatase has been assessed, by RT-PCR and immunocytochemistry, in the rat glioblastoma C6 and in two human glioblastoma cell lines T98G and U373MG. The three cell lines expressed aromatase mRNA and showed a cytoplasmic pattern of aromatase immunoreactivity. In addition, the three cell lines express estrogen receptor alpha, suggesting that estradiol formed by aromatase may act as an autocrine or paracrine factor for glioblastoma cells. By analogy to the implication of aromatase into the growth of other forms of estrogen-sensitive tumors, such as some breast cancers, it is conceivable that the expression of aromatase may play a role in the growth of glioblastomas. PMID- 15364412 TI - Plasmin deficiency does not alter endogenous murine amyloid beta levels in mice. AB - Deposition of amyloid beta (A beta) into extracellular plaques is a pathologic characteristic of Alzheimer's disease. Plasmin, neprilysin, endothelin-converting enzyme and insulin-degrading enzyme (IDE) have each been implicated in A beta degradation; data supporting the role of the latter three enzymes have included increased levels of endogenous murine A beta in mice genetically deficient for the respective enzyme. In this study, we sought to determine if plasminogen deficiency increases endogenous A beta. We report that plasminogen deficiency did not result in an A beta increase in the brain or in the plasma of adult mice. Hence, although plasmin is potentially important in the degradation of A beta aggregates, we interpret these data as suggesting that plasmin does not regulate steady-state A beta levels in non-pathologic conditions. PMID- 15364413 TI - Association study of three polymorphisms of kinesin light-chain 1 gene with Alzheimer's disease. AB - The transport of amyloid precursor protein is mediated through its interaction with kinesin light-chain 1 (KNS2). We hypothesized that kinesin light-chain dysfunction might be involved in the pathogenesis of Alzheimer's disease (AD). To assess the physiological relevance of an allelic variation in the KNS2 gene, the association analysis of three single nucleotide polymorphisms (SNPs) in the 5'UTR or in intronic sequences of KNS2 gene were performed in 100 AD brain patients and in 103 controls. For one of these polymorphisms (G58836C in intron 13), the association between AD and the C allele was found to be significant (odds ratio = 1.73, 95% CI: 1.12-2.67, P = 0.012). No synergistic effects were found between the APOE epsilon 4 allele and KNS2 gene polymorphisms. PMID- 15364414 TI - Acute restraint stress reduces protein kinase C gamma in the hippocampus of C57BL/6 but not DBA/2 mice. AB - Protein kinase C gamma (PKC gamma) is highly expressed in the rodent hippocampus and has been implicated in long-term alterations in synaptic efficacy. Acute stress has been shown to negatively affect hippocampal synaptic plasticity, and the present study examined the effect of acute stress on PKC gamma expression/subcellular distribution by quantitative western blotting in two inbred mouse strains (C57BL/6J versus DBA/2J) with established differences in hippocampal plasticity. It was found that both DBA/2J and C57BL/6J strains exhibited similar basal, stress-induced elevations, and recovery of serum corticosterone levels. Acute stress produced a significant reduction in both membrane and cytosolic PKC gamma expression in the hippocampus of C57BL/6J mice compared to no-stress controls, but did not alter either membrane or cytosolic PKC gamma expression in the hippocampus of DBA/2J mice compared to no-stress controls. These data provide direct evidence that PKC gamma is differentially regulated in the hippocampus of C57BL/6J and DBA/2J mice by acute stress. The role of stress-induced regulation of hippocampal PKC gamma expression in hippocampal synaptic plasticity is discussed. PMID- 15364415 TI - Noise-induced changes of neuronal spontaneous activity in mice inferior colliculus brain slices. AB - The inferior colliculus (IC) in vivo is reportedly subject to a noise-induced decrease of GABA-related inhibitory synaptic transmission accompanied by an amplitude increase of auditory evoked responses, a widening of tuning curves and a higher neuronal discharge rate at suprathreshold levels. However, other in vivo experiments which demonstrated constant neuronal auditory thresholds or unchanged spontaneous activity in the IC after noise exposure did not confirm those findings. Perhaps this can be the result of complex noise-induced interactions between different central auditory structures. It was, therefore, the aim of the present study to investigate the effects of noise exposure on the spontaneous electrical activity of single neurons in a slice preparation of the isolated mouse IC. Normal hearing mice were exposed to noise (10 kHz center frequency at 115 dB SPL for 3 h) at the age of 21 days under anesthesia (Ketamin/Rompun 10:1). After one week, auditory brainstem response (ABR) recordings and extracellular single-unit recordings from spontaneously active neurons within the IC slice were performed in noise-exposed and in normal hearing control mice. Noise-exposed animals showed a significant ABR threshold shift in the whole tested frequency range and a significant lower neuronal spontaneous activity in all investigated isofrequency laminae compared to controls. In both groups, the firing rate of 80% of IC neurons (approximately) increased significantly during the application of the GABA(A) receptor antagonist Bicucullin (10 microM). The present findings demonstrate a noise-related modulation of spontaneous activity in the IC, which possibly contribute to the generation of noise-induced tinnitus and hearing loss. PMID- 15364416 TI - Altered levels of plasma 24S- and 27-hydroxycholesterol in demented patients. AB - Alterations in brain cholesterol metabolism and reduced 24S-hydroxycholesterol plasma levels have been described in Alzheimer's disease (AD) and vascular dementia (VD). We hypothesize that changes in peripheral cholesterol metabolism, such as alterations in the plasma levels of 27-hydroxycholesterol, might also be involved. Plasma levels of 24S-hydroxycholesterol and 27-hydroxycholesterol in patients suffering from dementing disorders such as AD, VD, and mild cognitive impairment (MCI) were compared to those in age- and cholesterol matched non demented and depressed subjects. Cholesterol corrected concentrations of plasma 24S-hydroxycholesterol and 27-hydroxycholesterol were significantly reduced in patients with dementing disorders compared to non-demented subjects and depressed patients. A strong positive correlation between plasma 24S-hydroxycholesterol and 27-hydroxycholesterol levels was observed. The ratios of plasma 24S hydroxycholesterol to 27-hydroxycholesterol were higher in patients with dementing disorders compared to non-demented subjects. Our results support the observation, that cholesterol metabolism is altered in dementing disorders, indicated by different plasma concentrations of brain specific and peripherally produced oxysterols. PMID- 15364417 TI - Increased Alix (apoptosis-linked gene-2 interacting protein X) immunoreactivity in the degenerating striatum of rats chronically treated by 3-nitropropionic acid. AB - Chronic intoxication by 3-nitropropionic acid in the Lewis rat reproduces many features reminiscent of Huntington's disease including behavioural alterations and cortico-striatal degeneration. In particular, in this model, striatal degeneration is accompanied by calpain activation as found in the human disease. The present study was undertaken to determine whether the expression of Alix (apoptosis linked gene-2 interacting protein), a widespread protein involved in neuronal death, would be modified in the striatum and cortex of 3NP-treated rats. The results clearly show that Alix immunoreactivity is increased in the neuronal cell bodies of the lateral striatum, where degeneration is massive. The medial striatum and the cortex that lack neurodegeneration remain only weakly labelled. This is further evidence suggesting an involvement of Alix in the events driving neuronal death. PMID- 15364418 TI - Genetics, prefrontal cortex, and cognitive control: a twin study of event-related brain potentials in a response inhibition task. AB - Inhibition of prepotent responses plays a key role in cognitive executive control of goal-directed behavior. Response inhibition in Go/No-Go tasks elicits a distinct midfrontal ERP component, the N2, localized in previous studies to the anterior cingulate cortex. We assessed heritability of the No-Go N2 and the succeeding positive P3 component in 194 young female twins (52 monozygotic and 45 dizygotic pairs) who completed a cued version of the Continuous Performance Test. Genetic model-fitting analysis showed that about 60% of the variance in N2 and P3 amplitudes can be attributed to genetic factors, suggesting that frontal No-Go N2 and P3 components may index genetically transmitted individual differences in brain activity related to conflict processing and can potentially serve as endophenotypes for genetic studies of disorders characterized by neurobehavioral disinhibition. PMID- 15364419 TI - A novel presenilin 1 mutation (Y154N) in a patient with early onset Alzheimer's disease with spastic paraparesis. AB - Early onset familial Alzheimer's disease with spastic paraparesis (FAD-SP) has been associated with mutations of the presenilin 1 gene (PSEN1). We report a pedigree of FAD-SP due to a novel missense mutation of PSEN1 (Y154N). The symptoms of the proband were characterized by presenile dementia in her 40s, preceded by spastic paraparesis in her 30s, whereas the mother of the proband presented with spastic paraparesis in her 40s, followed by symptoms of dementia in her mid 60s. The mutation was found only in the proband, and not in a normal family member, normal Japanese control subjects, patients with sporadic Alzheimer's disease or patients with familial spastic paraparesis without dementia. Thus, Y154N is a novel PSEN1 mutation responsible for FAD-SP of Japanese origin. PMID- 15364420 TI - The KPNB3 locus is associated with schizophrenia. AB - Seven single nucleotide polymorphisms (SNPs) present on 13q32 were detected among 124 British family trios consisting of fathers, mothers and affected offspring with schizophrenia. The transmission disequilibrium test (TDT) demonstrated that of these 7 SNPs, rs626716, a T to C base change at the KPNB3 locus, was the only SNP associated with schizophrenia (chi(2) = 7.71, P = 0.005) although the global P-value given by a permutation test was 0.04 for 100 permutations. Of 248 parents, 20 were heterozygous. These heterozygous parents had transmitted 4 T alleles and 16 C-alleles to their affected offspring. To further validate the rs626716 association, we analysed a haplotype system composed of 3 SNPs at the KPNB3 locus. The result showed that the KPNB3 haplotypes were also associated with schizophrenia (chi(2) = 10.18, d.f. = 2, P = 0.006). Because the KPNB3 finding has been replicated in a Chinese population, it could be hypothesized that the KPNB3 locus may contain a disease-causing variant for schizophrenia. PMID- 15364421 TI - Ketamine potentiates the effect of electroacupuncture on mechanical allodynia in a rat model of neuropathic pain. AB - Mu-opioid agonists and N-methyl-d-aspartate (NMDA) receptor antagonists have been shown to attenuate mechanical allodynia in neuropathic pain models. We have previously reported that 2Hz electroacupuncture (EA) produced analgesia via releasing endogenous opioid peptides (i.e. beta-endorphin and endomorphin) and the activated micro-opioid receptors. The present study aimed to examine whether ketamine, an NMDA receptor antagonist, can enhance the anti-allodynic effects induced by 2Hz EA in a rat model of neuropathic pain following spinal nerve ligation (SNL). The results are as follows: (1) EA itself or i.p. injection of ketamine reduced mechanical allodynia (i.e. increase in withdrawal threshold). (2) Although injection of ketamine at a low dose (1.0mg/kg) alone did not influence mechanical withdrawal threshold, combination of ketamine at this dose with EA produced more potent anti-allodynic effect than that induced by EA alone. (3) The anti-allodynic effect of EA combined with ketamine could be reversed by i.p. injection of naloxone (2.0 mg/kg). These results suggested that ketamine potentiate the anti-allodynic of EA in rats with spinal nerve ligation, and endogenous opioid system is likely to be involved in this process. PMID- 15364422 TI - Magnetic resonance imaging of differential gray versus white matter injury following a mild or moderate hypoxic-ischemic insult in neonatal rats. AB - Selective white matter injury in the pre-mature infants suggests it has a greater susceptibility to hypoxia-ischemia. To investigate whether white matter injury would predominate following a mild hypoxic-ischemic insult, 7-day-old rats underwent either mild or moderate hypoxia-ischemia and magnetic resonance imaging 24 h later. Mild and moderate hypoxia-ischemia were produced by unilateral carotid artery occlusion plus exposure to hypoxia for either 45-50 or 90 min at ambient temperatures of 34.5 or 35.5 degrees C, respectively. Following mild hypoxia-ischemia, there was a significant increase in T(1) and T(2) within periventricular white matter (e.g. corpus callosum) in the hemisphere ipsilateral to the occlusion compared to that contralaterally and less of an increase within gray matter (e.g. cortex and striatum). This corresponded to relatively selective white matter injury detected histologically. Following a moderate hypoxia ischemia, both gray and white matter was severely injured with marked increases in T(1) and T(2) occurring in both white and gray matter regions ipsilateral to the hypoxia-ischemia. We conclude that a mild insult, consisting of a short duration of hypoxia-ischemia at a slightly lower body temperature than a moderate hypoxic-ischemic insult, produces enhanced injury in white matter and a relative sparing of gray matter. PMID- 15364423 TI - A transcranial magnetic stimulation study of the ipsilateral silent period in lower limb muscles. AB - The lower limb ipsilateral silent periods (ISP) were determined with transcranial magnetic stimulation in 30 normal subjects using a round coil. The mean duration and transcallosal conduction time were comparable to values obtained from upper limb recordings. No age-related correlation was found for either parameter, corroborating previous imaging and pathological studies of interhemispheric pathways. Our results highlight the feasibilty of eliciting ISPs in the lower limbs, relevant for future studies of interhemispheric interaction in clinical and research settings. PMID- 15364424 TI - Gabapentin relieves mechanical, warm and cold allodynia in a rat model of peripheral neuropathy. AB - Although recent studies demonstrated the relieving effect of gabapentin on neuropathic pain, the effect has not been sufficiently examined. In the present study, we investigated the effect of gabapentin on mechanical, warm and cold allodynia in a rat model of peripheral neuropathy. Under enflurane anesthesia, animals were subjected to the partial injury of the nerves innervating the tail. Behavioral tests for mechanical, cold and warm allodynia on the tail were performed by von Frey hair (2.0 g) stimulation, 4 and 40 degrees C water immersion, respectively. Intraperitoneal injection of gabapentin (30, 100, 300 mg/kg) significantly alleviated mechanical, warm and cold allodynia in a dose dependent manner. Our results suggest that gabapentin is an effective agent against mechanical, warm and cold allodynia in a rat model of peripheral neuropathy. PMID- 15364425 TI - Exposure to alcohol on E9 raises poststress corticosterone in mature but not old mice. AB - Prenatal exposure to alcohol alters postnatal function of the hypothalamic pituitary-adrenal axis. Hyperresponsiveness to stress, or increased secretion of corticosterone, is a commonly studied effect in offspring of rats exposed to alcohol during a substantial period of gestation. No studies have reported on stress hormone secretion following alcohol exposure on a single day during embryonic development even though exposure at this time may damage the hypothalamus and pituitary. To explore the effect of an acute exposure, we used the offspring of C57BL/6J mice exposed to alcohol or saline on embryonic day (E) 9 (2.9 g/kg administered twice, 4h apart). At 7.5 or 22 months of age these mice were subjected to a 12-h restraint stress, or merely kept in the same environment without restraint. After the 12-h period, a blood sample was obtained from the retro-orbital plexus, and analyzed for the amount of corticosterone. The 7.5 month old group of alcohol-treated offspring were indeed hyperresponsive to restraint stress, but the 22-month old mice were not. Whether the normal appearing corticosterone response of the old alcohol-exposed mice indicated adaptation to restraint, an aging-associated ceiling effect in corticosterone secretion, or an expression of pathology, cannot be decided on the basis of present data. PMID- 15364426 TI - Jerimiah Abalaka and HIV in Nigeria. PMID- 15364427 TI - Attempts to cure and prevent HIV/AIDS in central Nigeria between 1997 and 2002: opening a way to a vaccine-based solution to the problem? AB - Like other sub-Saharan countries Nigeria is ravaged by HIV/AIDS. The author is a general surgeon with some training in Immunology and practises in central Nigeria. He conceived and developed a safe therapeutic HIV vaccine prepared from the blood of HIV-infected persons and applied the remedy to over 3500 such patients with their informed and written consent. He also developed a safe HIV preventive vaccine from the same source and applied it to himself and over 300 willing HIV antibody negative persons. This work reports his efforts and the findings so far even though some of the patients involved are still undergoing the therapy. The therapy has not affected, to any significant statistical level, either the overall weight gain in all the patients or the CD4+ count in those patients that were able to pay for the tests. The therapy also resulted in a maintained seroreversion to negative (normal) in some of the patients who, prior to therapy, had established seropositivity for HIV antibodies (20 patients), hepatitis C virus (HCV) antibodies (16 patients) and hepatitis B surface antigen (HBsAg) (50 patients). Neither the therapeutic nor the preventive vaccine is associated with the production of anti-HIV antibodies. PMID- 15364428 TI - Need for re-appraisal of acute flaccid paralysis (AFP) case classification. PMID- 15364429 TI - Pre-clinical evaluation of the malaria vaccine candidate P. falciparum MSP1(42) formulated with novel adjuvants or with alum. AB - We compared the safety and immunogenicity of the recombinant Plasmodium falciparum MSP1(42) antigen formulated with four novel adjuvant systems (AS01B, AS02A, AS05 and AS08) to alum in rhesus monkeys. All five formulations of MSP1(42) were safe and immunogenic. Whereas, all MSP1(42) formulations tested generated high stimulation indices for lymphocyte proliferation (ranging from 27 to 50), the AS02A and AS01B formulations induced the highest levels of specific anti-MSP1(42) antibody. ELISPOT assays showed that the AS02A and AS01B vaccine formulations-induced different cytokine response profiles. Using the ratio of IFN gamma/IL-5 secreting cells as the metric, the AS01B formulation induced a strong Th1 response, whereas the AS02A formulation induced a balanced Th1/Th2 response. The IFN-gamma response generated by AS02A and AS01B formulations persisted at least 24 weeks after final vaccination. The notable difference in Th1/Th2 polarization induced by the AS02A and AS01B formulations warrants comparative clinical testing. PMID- 15364430 TI - Protection of pigs against Taenia solium cysticercosis using recombinant antigen or in combination with DNA vaccine. AB - In the present study, we investigated the duration of protection afforded to pigs immunized in two different prime-boost regimens: one is homologus priming and boosting with a protein vaccine, and the other is priming with a DNA vaccine and boosting with the protein vaccine. Groups of pigs that received the same vaccination regimen were then challenged with Taenia solium eggs at 6, 12 or 20 weeks post-immunization (wpi), respectively. The results showed that all vaccinated pigs challenged at 6 or 12 wpi showed significant (P < 0.05) reduction in the development of cysts. When challenged at 20 wpi, pigs primed with the DNA vaccine (pcDNA3-cC1) followed by two boosters of the protein vaccine (GST-cC1) showed significant (P < 0.05) protection against the challenge of T. solium eggs, whereas pigs receiving three injections of the protein vaccine showed no significant protection compared to non-vaccinated controls (P > 0.05). Antibody isotype assays showed that DNA prime-protein boost regimen induced a predominantly IgG2 response, compared to an IgG1 biased response for the protein prime-protein boost regimen. In addition, peripheral blood mononuclear cells (PBMC) obtained from the DNA prime-protein boost group proliferated strongly in response to GST-cC1 protein, and this responsiveness persisted until 20 wpi. Taken together, our data suggest that the use of a prime-boost strategy combining DNA and protein vaccines may be better than protein alone for the longevity of protection against the challenge of T. solium eggs. PMID- 15364431 TI - Bacillus Calmette-Guerin shares with virulent Mycobacterium tuberculosis the capacity to subvert monocyte differentiation into dendritic cell: implication for its efficacy as a vaccine preventing tuberculosis. AB - The only available vaccine against tuberculosis (TB) is Bacillus Calmette-Guerin (BCG) whose efficacy in preventing pulmonary tuberculosis is however controversial. Here, we show that BCG infection of monocytes causes their differentiation into mature dendritic cells (DCs) lacking CD1 molecules expression, coupled with suboptimal up-regulation of HLA class II, CD80 and CD40 molecules and a marked unresponsiveness to lipopolysaccharide stimulation. In addition, alloreactive naive T lymphocytes primed by these subverted DCs did not undergo defined functional polarization, as witnessed by their inability to produce IFN-gamma. Since efficient antigen presentation and IFN-gamma production by mycobacterial-specific T lymphocytes are required for protection against Mycobacterium tuberculosis, our data might provide additional explanation for the low efficacy of BCG vaccination. PMID- 15364432 TI - Optimization of microcarrier cell culture process for the inactivated enterovirus type 71 vaccine development. AB - Enterovirus 71 (EV71) is an enterovirus that could lead to severe neurological disorders and fatalities. The inactivated vaccine is an appropriate EV71 vaccine format for meeting current needs. Large-scale preparation of the inactivated EV71vaccine depends on a scalable cell culture system for industrial mass production. In this paper, Vero cells were found to produce higher titers of EV71 than did MRC-5 and WI-38 cells. High-density microcarrier Vero cell cultures were established using 5g/L Cytodex 1 microcarriers and found to promote the release of EV71s from infected Vero cells. For the large-scale production of the inactivated vaccine antigen, the extracellular virus titers produced in the 2L bioreactor were found to be 10 times lower than the spinner flask culture but improved by 30-folds using glucose/glutamine feedings during infection. A serum free Vero cell microcarrier culture was also established in the bioreactor, yielding a high-titer of 5.8 x 10(7) TCID50/mL for EV71 production. The immunogenicity of the inactivated virions produced in serum-free culture elicited a slightly higher level of neutralizing antibody response in immunized mice. These results constitute valuable information on the development of a large-scale microcarrier cell culture process for producing inactivated EV71 vaccine. PMID- 15364433 TI - Vaccination with a plasmid DNA cocktail encoding the nucleosomal histones of Leishmania confers protection against murine cutaneous leishmaniosis. AB - Leishmania histones are relevant immunogens for the host immune system during both Leishmania infection and disease. In the present paper we have evaluated the prophylactic value of the four Leishmania infantum histones forming the nucleosomal core in the murine model of cutaneous leishmaniasis. In a first stage, the immune response elicited by the intramuscular injection of a mixture of four plasmid DNAs, encoding the L. infantum histones H2A, H2B, H3 and H4, was determined in BALB/c mice. It was found that the immunized animals developed a specific Th1 immune response, which was associated with an antigen-specific production of interferon (IFN-gamma) and a limited humoral response against histones (dominated by antibodies of the IgG2a isotype). According to the pure Th1-type immune response elicited by the DNA vaccination with Leishmania histones, vaccinated mice showed a solid immunity that efficiently controlled the Leishmania major infection. The protection in mice vaccinated with histone-DNAs was associated with a low humoral response against leishmanial antigens, an enhanced IFN-gamma production and little, if any, IL-4 production. The relative contribution of both CD8(+) and CD4(+) T cells to the IFN-gamma production, and the IL-12 dependence were also evaluated. All these data indicated that DNA vaccination with Leishmania histones genes results in a specific Th1-like response during L. major infection, and that both CD4(+) and CD8(+) T cells contribute to the resistance of vaccinated mice to cutaneous leishmaniasis. PMID- 15364434 TI - Long-term effects of vaccination on attentional performance. AB - To investigate the possible influence of stimulation of the immune system on cognitive tasks, healthy volunteers were vaccinated against hepatitis B and tested over a 6 month-period in a simple reaction times and the Stroop task. In general, the "Stroop effect" demonstrates that both the name and meaning of a word are automatically processed even when voluntary attention is trying hard not to process them. Unlike placebo group, vaccinated subjects showed a persistent lack of the classical Stroop effect. These findings may be explained by a constraint satisfaction model of the Stroop task, assuming a selective weakening of the connection matrix, and suggest that immune-cognitive effects may occur, besides the well known immune-cognitive influences like those elicited by emotional stress. PMID- 15364435 TI - Adjuvant effect of Panax notoginseng saponins on the immune responses to ovalbumin in mice. AB - In this study, the haemolytic activities of Panax notoginseng saponins (PNS) and its adjuvant potentials on the cellular and humoral immune responses of ICR mice against ovalbumin (OVA) were evaluated. We determined the haemolytic activity of PNS using 0.5% rabbit red blood cell. PNS showed a slight haemolytic effect, with its haemolytic percents being 11.59 and 3.60% at the concentrations of 500 and 250 microg/ml, respectively. Furthermore, the adjuvant potential of PNS at three dose levels on the cellular and humoral immune responses of ICR mice against ovalbumin were investigated. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing aluminum hydroxide gel (Alum) (200 microg), Quil A (10 and 50 microg) or PNS (50, 100 or 200 microg) on days 1 and 15. Two weeks later (day 28), concanavalin A (Con A)-, pokeweed (PWM)- and OVA-stimulated splenocyte proliferation and OVA-specific antibodies in serum were measured. PNS significantly enhanced the Con A-, PWM-, and OVA-induced splenocyte proliferation in the OVA-immunized mice at a dose of 100 microg (P < 0.05 or P < 0.025). OVA-specific IgG, IgG1 and IgG2b antibody levels in serum were significantly enhanced by PNS compared with OVA control group (P < 0.025). Moreover, enhancing effect of PNS on the OVA-specific IgG2b antibody responses to OVA in mice were more significant than that of Quil A (P < 0.025). In conclusion, the results suggest that PNS could be safely used as adjuvant with low or non-haemolytic effect. PMID- 15364436 TI - Typing of pneumococcal surface protein A (PspA) in Streptococcus pneumoniae isolated during epidemiological surveillance in Brazil: towards novel pneumococcal protein vaccines. AB - Pneumococcal protein vaccine based on pneumococcal surface protein A (PspA) is in development with the potential to offer broad range of protection against different strains. We have investigated the frequency of PspA family 1 (Fam1) and family 2 (Fam2) proteins among Streptococcus pneumoniae recovered from ongoing surveillance in Brazil. Fam1 and Fam2 were expressed in comparable rates among 366 isolates, with the potential coverage of 94.3%. PspA families were not associated to age group or source of isolates. However, considering the significant tendency of increasing prevalence of Fam2 associated to widespread dissemination of the genetically-related resistant strains, the monitoring of the PspA families derived from population-based data may be necessary in the context of vaccine development. PMID- 15364437 TI - Hepatitis B vaccination in prison with a 3-week schedule is more efficient than the standard 6-month schedule. AB - A randomized study of injecting drug users in a Danish prison comparing vaccination at 0, 1 and 3 weeks with the 0, 1 and 6 months schedule (20microg Engerix B i.m.) was conducted. Due to a low participation rate, a second nonrandomized study was conducted in Estonia where all prisoners were vaccinated with the short schedule. In the randomized study, the compliance with three doses was 63% (12/19) in the 3-week schedule compared to 20% (3/15) in the 6-month schedule (P = 0.017). In the nonrandomized study, the compliance was 81% (457/566) and the seroprotection rate at month 7 was 67% (97/145), resulting in protection of 54% of the population at risk. This was significantly higher than the estimated 34-42% protection that would have been achieved with the 6-month schedule. PMID- 15364438 TI - Continuous antigenic stimulation system (CASS) as a new immunization strategy. AB - Protection against diseases is mediated by a sustained immune response. Here, we describe a new immunization strategy. Mice implanted with encapsulated C2C12 myoblasts secreting human factor IX (hFIX) elicited a strong humoral response against the transgene, as compared to mice immunized with complete Freund's adjuvant (FA). Mice also had increasing IgG2a antibody titer, indicating a switch to a Th1 profile immune response. Mice developed strong hFIX-specific cytotoxic T lymphocytes (CTL) that was detectable 213 days after implantation, demonstrating the sustained immunity elicited by encapsulated cells. Here, we propose continuous antigenic stimulation system (CASS) as a novel immunization strategy with potential application in the design of novel vaccines. PMID- 15364439 TI - Safety concerns regarding combination vaccines: the experience in Japan. AB - This study explored the safety concerns associated with combination vaccines in Japan. The impact of Japan's decision in 1975 to withdraw the combined diptheria, tetanus, and pertussis (DTP) vaccine, and then in 1993, the combined measles, mumps, and rubella (MMR) vaccine and provide only the single antigen vaccines was analyzed. Interviews with both governmental and non-governmental agency officials in Japan demonstrated that withdrawal of the DTP and MMR vaccines had significant impact on the rates of immunization and disease despite the availability of monovalent vaccines. PMID- 15364440 TI - Comparative immunogenicity analysis of modified vaccinia Ankara vectors expressing native or modified forms of hepatitis C virus E1 and E2 glycoproteins. AB - We have evaluated in C57/Bl6 and HLA-A2.1 transgenic mice the immunogenicity of three MVA vectors expressing either native HCV E1E2 polyprotein, truncated and secreted E1 (E'1(311)) and E2 (E'2(661)) proteins, or a chimeric E1E2 heterodimer presented at the plasma membrane. Immunization induced mainly a Th1 response in HLA-A2.1 transgenic mice while a Th2-type response was detected in C57/Bl6 mice. Comparison of the three vectors shows an increase in the humoral response when antigens are secreted or membrane bound, and slightly in the cellular response when antigens are exposed on the cell surface. PMID- 15364441 TI - Intranasal immunisation with Toxoplasma gondii tachyzoite antigen encapsulated into PLG microspheres induces humoral and cell-mediated immunity in sheep. AB - Proteins from a crude extract of Toxoplasma gondii tachyzoites were encapsulated into poly(D,L-lactide-co-glycolide) (PLG) micro- and nano-particles with a mean encapsulation efficiency of 80%. An intranasal immunisation and infection experiment using 24 sheep was conducted to compare the immune responses elicited by intranasal administration of soluble and particulate T. gondii antigen (with and without cholera toxin). Sheep immunised with particulate toxoplasma antigen produced enhanced levels of both local and systemic antigen-specific IgA antibody, and showed increased cellular immune responses with a corresponding increase in IFNgamma production. After challenge with toxoplasma oocysts larger quantities of both nasal and systemic IgG were measured more rapidly in all animals immunised with toxoplasma antigen than animals infected with oocysts, suggesting a secondary-type IgG response. A slight modification of the febrile response to toxoplasma infection could be observed in animals immunised with particulate toxoplasma antigen and cholera toxin, although none of the immunised animals were protected against the challenge infection. These studies show that intra-nasal delivery has the potential to be an effective route for mucosal immunisation in sheep. PMID- 15364442 TI - Efficacy of DNA vaccines expressing the type F botulinum toxin Hc fragment using different promoters. AB - DNA vaccines which expressed the Hc fragment of the Clostridium botulinum type F neurotoxin (BoNT/F Hc) fused to a signal peptide downstream of four different eukaryotic promoters were prepared. Subsequently, the immunogenicity of the DNA vaccines and protection afforded in mice against challenge with 10(4) MLD of type F botulinum toxin was evaluated. The DNA vaccine containing the human ubiquitin gene (UbC) promoter induced the highest BoNT/F Hc-specific antibody concentration following two intramuscular immunisations and afforded 90% protection against challenge. The results from this study indicate that the selection of promoter used in DNA vaccination studies may be of importance in designing optimised vaccines. PMID- 15364443 TI - Epidemiology of severe varicella-zoster virus infection in Spain. AB - Data of hospitalizations for varicella and herpes zoster in Spain during the 1999 2000 period were obtained from the national surveillance system for hospital data. A total of 3083 hospitalizations for varicella and 6324 for herpes zoster were identified, representing an annual incidence of 4.1 and 8.4 per 100,000 persons per year, respectively. Almost half of patients hospitalized for varicella were children under 5 years of age. In contrast, 78% of hospitalizations for zoster occurred in adults >50 years of age. Hospitalizations for varicella and herpes zoster resulted annually in 11,141 and 40,090 days of hospitalization and a cost of 3.2 and 7.0 million, respectively. PMID- 15364444 TI - Incidence of Haemophilus influenzae type b and other invasive diseases in South Korean children. AB - To determine incidence of invasive Haemophilus influenzae type b (Hib) disease in a defined population of Jeonbuk Province, Korea, children <5 years were evaluated in prospective, population-based surveillance of invasive bacterial diseases using standardized methods for patient referral, clinical evaluation and laboratory testing (optimized culture, latex agglutination, polymerase chain reaction). Vaccine utilization was assessed with vaccination histories of patients in surveillance, monthly data on Hib vaccine distribution and a coverage survey of clinic patients in study population. From September 1999 to December 2001, 2176 children were evaluated for possible meningitis, 1541 had no cerebrospinal fluid (CSF) findings of meningitis, 605 had CSF abnormalities (suspected bacterial meningitis) but no pathogen identified; six patients had probable Hib meningitis and eight had confirmed Hib meningitis. The annual suspected bacterial meningitis incidence was 258.4/100,000 <5 years and the probable/confirmed Hib meningitis incidence was 6.0/100,000 <5 years. Pneumococcal meningitis incidence was 2.1/100,000 <5 years and Group B streptococcal meningitis incidence was 0.17/1000 live births. A total of 69,589 Hib vaccine doses were distributed during the study. Hib vaccine coverage was negligible initially but increased to 16% (complete Hib immunization) and 27% (partial immunization) in final months of study. Suspected bacterial meningitis incidence was high but proven invasive Hib meningitis incidence was low. Hib was leading cause of bacterial meningitis yet bacterial pathogens were identified in only 4% of abnormal CSF. These findings may reflect truly low incidence, presumptive antibiotic treatment, partial Hib immunization, or incomplete clinical evaluations. Given the apparent Hib meningitis burden in Jeonbuk Province, additional studies to describe other invasive Hib syndromes, Hib associated mortality and disability, and economic impact of Hib disease will be useful to guide public health decisions regarding routine Hib vaccine introduction. PMID- 15364445 TI - The evaluation of immune responses that occur after HBV infection and HBV vaccination in hemodialysis patients. AB - The hepatitis B virus (HBV) vaccination never lose its importance; however, we did not get immune response with vaccination in some cases at the hemodialysis (HD) units. In this research, our aim was to evaluate antibody responses that constitute with natural ways and vaccination against HBV infection in chronic HD patients. In this retrospective cohort study (between 1999 February and 2001 December), 34 patients, 19 males and 15 females that were at the HD program, were enrolled. Patients were divided into two groups: group A, vaccination group (n = 15); and group B, anti-HBc IgG (+) cases (n = 19). We injected 40 microg of recombinant HBV vaccine into the deltoid muscle at 0, 1, 2, and 6 months. The anti-HBs levels, over 10 mIU/mL, were accepted for enough immunization. In group A, after vaccination 12 of 15 (80%) patients had seroconversion but later, in five of 12 (33.3%) patients, HBs-antibody became negative and in seven of 12 (46.7%) patients the positivity of HBs-antibody continued. However, in group B, 16 of 19 (66.7%) immune patients who faced HBV infection, it was established that antibody response continued (P > 0.05). The patients in group A who had anti-HCV negativity and serum albumin levels >/=3.5g/dL (7/0; P < 0.05) had too much antibody response against the vaccine. The difference of our research was that patients under 49 years old, patients who did not respond to HBV vaccine, were proportionally higher as compared to those from other researches. In conclusion, it was realized that the immune response of the HBV vaccine was low in the HD patients and it was affected by several factors such as gender, anti-HCV positivity and nutritional status. But, in spite of getting all universal precautions, still to face with new cases emphasizes the importance of the vaccination. PMID- 15364446 TI - Association of antigens to ISCOMATRIX adjuvant using metal chelation leads to improved CTL responses. AB - The association of antigen with ISCOMATRIX trade mark adjuvant has been shown to be important for the optimal induction of cytotoxic T lymphocyte (CTL) responses. Here, we describe a simple broadly applicable method for associating recombinant proteins with hexa-histidine tags to ISCOMATRIX trade mark adjuvant utilising metal-affinity chelating interactions. The metal chelation binding step can be performed in a wide range of buffers, including commonly used denaturants such as urea, which makes it an ideal strategy for formulating proteins which are otherwise insoluble. Following association of protein with the chelating ISCOMATRIX trade mark adjuvant, the denaturant can be removed. Further, we show enhanced CTL responses with a protein-associated chelating ISCOMATRIX trade mark vaccine compared to a non-associated ISCOMATRIX trade mark vaccine. PMID- 15364447 TI - Reducing methane emissions in sheep by immunization against rumen methanogens. AB - This work was conducted to determine if methane emissions from sheep immunized with an anti-methanogen vaccine were significantly lower than methane emissions from non-immunized sheep, to test the effectiveness of two different vaccine formulations (VF) on methane abatement, and to compare methane emissions measured using a closed-circuit respiration chamber and the sulphur-hexafluoride (SF6) tracer technique. Thirty mature wether sheep were randomly allocated to three treatment groups (n = 10). One group received an immunization of adjuvant only on days 0 and 153 (control), a second group received an immunization with a 3 methanogen mix on days 0 and 153 (VF3 + 3), and a third group received an immunization of a 7-methanogen mix on day 0 followed by a 3-methanogen mix on day 153 (VF7 + 3). Four weeks post-secondary immunization, there was a significant 7.7% reduction in methane production per kg dry matter intake in the VF7 + 3 group compared to the controls (P = 0.051). However, methane emissions from sheep immunized with VF7 + 3 were not significantly different when compared to the sheep in the control group (P = 0.883). The average IgG and IgA antibody titres in both plasma and saliva of the VF3 + 3 immunized sheep were four to nine times higher than those immunized with VF7 + 3 (P< 0.001) at both 3 and 6 weeks post secondary immunization. Data also revealed that SF6 methane estimates were consistently higher than the respiration chamber estimates and that there was no significant correlation between the SF6 methane estimates and the respiration chamber methane estimates (R2 = 0.11). PMID- 15364448 TI - Enhancement of vaccine-specific cellular immunity in infants by passively acquired maternal antibody. AB - The known protective effects of passively acquired maternal antibody on the resistance of newborns to infections have prompted widespread interest in maternal vaccination. However, a range of animal model and human studies indicate potential inhibitory effects of maternal antibody on vaccine-specific humoral responses in infants. In the present study we have examined the relationship between maternally acquired TT-specific IgG present before DTaP vaccination and subsequent TT-specific T-cell memory responses at 12 and 18 months, in a cohort of 118 infants. We demonstrate a strong positive association between TT-specific cellular immunity as evidenced by increased IL-4, IL-5 and IL-13 responses, and maternal TT-specific IgG. PMID- 15364449 TI - Enhancement of vaccinia vaccine potency by linkage of tumor antigen gene to gene encoding calreticulin. AB - Vaccinia vaccines have become important vectors for antigen-specific immunotherapy. Calreticulin has been shown to enhance MHC class I presentation of linked peptide/protein and may be useful for antigen-specific cancer treatment. An innovative vaccine administering antigen linked to calreticulin via a vaccinia vector may generate a potent antigen-specific antitumor response. We tested the efficacy of linking calreticulin (CRT) to model antigen human papilloma virus type 16 (HPV-16) E7 in the context of a vaccinia vaccine (Vac-CRT/E7). Intraperitoneal vaccination of C57BL/6 mice with Vac-CRT/E7 led to a dramatic increase in E7-specific IFN-gamma-secreting CD8+ T cells and a potent antitumor effect against E7-expressing tumors compared to immunization with Vac-E7 or Vac CRT. When compared to other chimeric vaccinia vaccines employing various intracellular targeting strategies previously developed in our lab, Vac-CRT/E7 elicited the highest number of E7-specific CD8+ T cells. Thus, vaccination with vaccinia expressing CRT linked to a tumor antigen may represent an advantageous strategy for cancer immunotherapy. PMID- 15364450 TI - Critical prevalence of antibodies minimizing vaccination costs for hepatitis A, hepatitis B, varicella, measles and tetanus in adults and adolescents in Catalonia, Spain. AB - The prevalence of seropositive individuals that makes costs of vaccinating all individuals equal to that for screening and vaccination of susceptible individuals is defined as the critical prevalence of antibodies (p*). Screening and vaccination is more efficient when the prevalence of seropositive individuals (p) in the population is higher than p*. In this study, the formula to obtain p* was derived from the cost-effectiveness equations, showing that it depends on screening and vaccination costs, programme compliance, screening test performance, vaccine efficacy and disease costs. The formula was used to determine the least costly vaccination strategy for hepatitis A and B, varicella, measles and tetanus in adults and adolescents in Catalonia. The least costly vaccination strategy was vaccination without screening (since p was lower than p*) for hepatitis B, measles and tetanus in adults and adolescents (5-14 years) and for hepatitis A in individuals aged 5-24 years, and screening and vaccination (since p was higher than p*) for varicella in adults and adolescents and for hepatitis A in adults aged >24 years. Vaccination strategies based on the critical prevalence of antibodies could maximize the immunity level in the community from available resources. PMID- 15364451 TI - Multiplex opsonophagocytosis assay (MOPA): a useful tool for the monitoring of the 7-valent pneumococcal conjugate vaccine. AB - Pneumococcal conjugate vaccination is highly efficacious against invasive diseases in young children. Since host protection is mainly mediated by opsonin dependent phagocytosis, the in vitro measurement of opsonophagocytic activity of the anti-capsular antibodies is assumed to be a reliable correlate of protection to monitor vaccine efficacy. Unfortunately, the methods used so far are all tedious to perform and material-consuming. Therefore, we modified the multi specificity opsonophagocytosis killing assay (MSOPKA) into a high-throughput method, which simultaneously measures the opsonophagocytosis against the seven serotypes covered by the current conjugate vaccine in a single assay. In the so called multiplex opsonophagocytosis assay (MOPA), a mixture containing equal numbers of colony forming units (CFUs) of chloramphenicol-resistant serotype 4, spectinomycin-resistant serotype 6B, streptomycin-resistant serotype 9V, erythromycin-resistant serotype 14, rifampicin-resistant serotype 18C, tetracycline-resistant serotype 19F, and trimethoprim-resistant serotype 23F pneumococci was used as a target mixture and incubated with serial dilutions of test serum. After opsonophagocytosis by differentiated HL-60 cells in the presence of complement, the samples were spotted onto different blood agar plates containing the seven selective antibiotics, respectively. Opsonophagocytosis was calculated as the highest serum dilution resulting in 90% or more reduction in CFUs. The data obtained by this assay correlated well with the data obtained by the MSOPKA. In conclusion, the MOPA simultaneously measures opsonophagocytosis capacity of serum against the capsular serotypes included in the 7-valent pneumococcal conjugate vaccine in a high-throughput fashion, requiring low volumes of patient sera. PMID- 15364452 TI - Intranasal immunisation of mice with liposomes containing recombinant meningococcal OpaB and OpaJ proteins. AB - The opacity (Opa) proteins of Neisseria meningitidis are outer membrane proteins involved in adhesion and invasion of host epithelial cells and are therefore expected to play an important role in colonisation of the nasopharynx. The majority of meningococcal Opa proteins bind to members of the CEACAM receptor family, such as CEA. Blocking of the Opa-CEACAM interaction by mucosal anti-Opa antibodies could thus constitute an important protective mechanism for novel meningococcal vaccines. In this study we analysed the specific anti-Opa antibody responses after intranasal immunisation of mice with liposomes containing purified and native OpaB (recognising the CEA receptor) and OpaJ (no affinity for CEA) proteins. These antigens were combined with or without one of three different adjuvants, i.e. purified meningococcal LPS, monophosphoryl lipid A (MPL) or the B-subunit of Escherichia coli heat-labile enterotoxin (EtxB). After intranasal immunisation with any of these formulations, anti-Opa IgA antibodies were found in nasal lavages and in some cases anti-Opa IgA and IgG antibodies were also found in lung lavages. With OpaJ but not OpaB, significant bactericidal serum titres were obtained. Of the different adjuvants used, meningococcal LPS gave the strongest overall immune response. Non-adjuvated liposomal Opa formulations were poorly immunogenic. No differences were found between the immune response in transgenic mice expressing the CEA-receptor and non-transgenic mice, showing that the CEA-Opa interaction does not influence the antibody response. PMID- 15364453 TI - Vaccination of C57/BL6 mice with Dobrava hantavirus nucleocapsid protein in Freund's adjuvant induced partial protection against challenge. AB - Dobrava hantavirus (DOBV) causes a severe form of hemorrhagic fever with renal syndrome (HFRS) for which there is no therapy or vaccine available. We compared the immunogenicity and protective efficacy of recombinant DOBV nucleocapsid protein (rDOBV N) given with Alum or Freund's as adjuvant, or PBS, in C57/BL6 mice. All mice given Alum or Freund's seroconverted as did 6/8 mice given rDOBV N with PBS. Reciprocal geometric mean total IgG-titers were 5380, 18,100, and 800, respectively, while the mean IgG1/IgG2a ratios were 17.5, 9.25, and 12, respectively. Furthermore, ELIspot assays showed higher levels of IL-4 producing peripheral blood mononuclear cells (PBMCs) in the group given Alum as compared to the other groups. Interestingly, only mice receiving rDOBV N with Freund's adjuvant were protected from challenge (75% protected), indicating that the strong Th2-type of immune response induced by Alum against rDOBV N did not induce protection in mice. PMID- 15364454 TI - Recombinant vesicular stomatitis (Indiana) virus expressing New Jersey and Indiana glycoproteins induces neutralizing antibodies to each serotype in swine, a natural host. AB - Vesicular stomatitis virus (VSV) is the most common cause of vesicular disease outbreaks in livestock throughout the Western Hemisphere. Two major serotypes, Indiana and New Jersey, cause epidemic disease in pigs, cattle, and horses. We generated recombinant viruses derived from the Indiana serotype genome that were engineered to contain and express: (1) a single copy of the glycoprotein gene from the Indiana serotype (VSIV-GI); (2) a single copy of the glycoprotein gene from the New Jersey serotype (VSIV-GNJ); or (3) two copies of the glycoprotein gene, one from each of the two major VSV serotypes (VSIV-GNJGI) [Martinez I, Rodriguez LL, Jimenez C, Pauszek SJ, Wertz GW. Vesicular stomatitis virus glycoprotein is a determinant of pathogenesis in swine, a natural host. J Virol 2003;77(14):8039-47]. These recombinant viruses and a highly pathogenic New Jersey field isolate (VSNJV), from which the GNJ gene was derived, were inoculated into swine, a natural host, and the induction of neutralizing antibodies to both serotypes was analyzed. The neutralizing antibody response induced by VSIV-GI, VSIV-GNJ and VSNJV was serotype-specific, according to the glycoprotein expressed. VSIV-GNJGI expressed both glycoproteins stably through multiple rounds of replication in swine and induced neutralizing antibodies against both VSV serotypes, with a dominance of the Indiana serotype in the serological response. Pigs immunized with VSIV-GI or VSIV-GNJ were protected against homologous high dose virus challenge. Pigs inoculated with VSIV-GNJGI were protected against challenge with VSIV-GI but three of four animals developed lesions after challenge with the highly pathogenic New Jersey field isolate. PMID- 15364455 TI - A therapeutic HIV vaccine: how good is good enough? AB - The goal of a therapeutic HIV vaccine is to attenuate HIV disease progression in those already infected. Our objective was to establish comparative efficacy and cost-effectiveness thresholds at which a therapeutic vaccine would make a valuable contribution to HIV care. Using an HIV computer simulation model, we compared therapeutic vaccination with HIV standard of care without vaccination. Input data were obtained from the literature. Base case and sensitivity analyses related to vaccine magnitude, penetrance, durability, and cost. In the base case (0.5 log magnitude, 25% penetrance, 3-year durability, and US$ 4000 per series), vaccination increased quality-adjusted life expectancy (QALE) by 0.50 months compared to no vaccination (cost-effectiveness ratio US$ 89,900 per quality adjusted life year (QALY)). Increasing vaccine penetrance to 50% increased the projected QALE benefit to 0.91 months (cost-effectiveness ratio US$ 45,500/QALY). Even modestly effective therapeutic HIV vaccines may produce small but meaningful increases in life expectancy and compare favorably to alternative uses of scarce HIV care resources. PMID- 15364456 TI - Vaccination protects B cell-deficient mice against an oral challenge with mildly virulent Toxoplasma gondii. AB - Prior studies have demonstrated that B cells are important components of protection in vaccinated mice challenged intraperitoneally with a highly virulent type I strain of Toxoplasma gondii parasites. However, it is not known whether B cells are required for vaccinated mice to successfully resist a more physiologically relevant challenge infection with a mildly virulent type II strain of T. gondii. To investigate that question, we vaccinated B cell-deficient C57BL/6 (microMT) mice with an attenuated strain of T. gondii and challenged them with a potentially lethal oral dose of type II T. gondii cysts. Vaccinated microMT mice survived the challenge as well as vaccinated B6 controls, controlled parasites equally well in critical tissues, produced equivalent levels of mRNA for several type 1 cytokines, and exhibited comparably mild histopathology. Thus, a vaccine can protect against infection with a mildly virulent type II strain of T. gondii in the absence of a B cell-dependent immune response. PMID- 15364457 TI - Efficacy of immune sera from human immunoglobulin transgenic mice immunized with a peptide mimotope of Cryptococcus neoformans glucuronoxylomannan. AB - The efficacy of antibody mediated immunity against Cryptococcus neoformans has not been established experimentally for human antibodies. Our group has previously shown that immunization with a conjugate consisting of a peptide mimotope of the C. neoformans capsular polysaccharide glucuronoxylomannan (GXM), P13, and diphtheria toxoid (P13-DT) prolonged survival of transgenic mice with human immunoglobulin loci, XenoMouse mice, which were challenged with a lethal dose of C. neoformans. In the study reported herein, we determined the efficacy of human antibodies in the sera of immunized XenoMouse mice against C. neoformans in passive transfer experiments in naive BALB/c mice. Survival studies were performed with sera from XenoMouse mice expressing human IgG2/kappa (G2/k mice) or IgG4/kappa (G4/k mice) that had been immunized with P13-tetanus toxoid (TT)/Alhydrogel with or without CpG, and G2/k mice that had been immunized with P13-DT/Alhydrogel/CpG or Alhydrogel/CpG, obtained on day 7 (early sera) and days 30 or 35-59 (late sera) after primary immunization. Compared to mice receiving sera from G2/k-PBS-treated mice, the survival of naive mice was prolonged by both early and late sera from G2/k-P13-DT/Alhydrogel/CpG-immunized mice, but only late sera from G2/k-P13-TT/Alhydrogel/CpG-immunized mice. Late, but not early sera from G2/k-Alhydrogel/CpG-immunized mice also prolonged survival. For all sera, prolongation of survival was associated with GXM-specific serum IgM. Sera from G2/k mice that received P13-TT without CpG, and all groups of G4/k mice had low to undetectable levels of antibody to GXM and were not protective. Our findings suggest that GXM-specific human IgM may be a functional mediator of protection against C. neoformans. PMID- 15364458 TI - Cloning of the varicella-zoster virus genome as an infectious bacterial artificial chromosome in Escherichia coli. AB - The complete genome of the varicella-zoster virus (VZV) Oka strain has been cloned as a bacterial artificial chromosome (BAC). Following electroporation into Escherichia coli (E. coli) strain DH10B, the VZV BAC was stably propagated over multiple generations of its host. Human embryonic lung (HEL) cells transfected with VZV BAC DNA recovered from DH10B showed cytopathic effect (CPE), and virus spread to neighbouring cells was observed. BAC vector sequences are flanked by loxP sites and, coinfection of the reconstituted virus, with a recombinant adenovirus expressing Cre recombinase removed the bacterial sequences. The resulting recombinant rV02 grew as well as the parental virus in HEL cells. The recombinant VZV will promote VZV research and increase use of the viral genome as an investigative tool. PMID- 15364459 TI - Differences in the immune response to long term Abeta vaccination in C57BL/6 and B6D2F1 mice. AB - The cerebral accumulation of beta-amyloid (Abeta) is a pathological hallmark of Alzheimer's disease (AD). Abeta vaccination or anti-Abeta specific antibodies may be a possible therapeutic option for AD. Previously, we demonstrated variation in the humoral response between B6D2F1 and C57BL/6 during short term (14 weeks) Abeta immunization. In the present study, we determined the humoral and cellular immune responses in these same mouse strains to a longer period of Abeta vaccination and further refined the major B cell epitope to Ass1-7. B6D2F1 mice generated a greater humoral and Th1 immune response versus C57BL/6 mice. Immunization with 25 microg Abeta produced a greater T cell response in B6D2F1 mice compared to 50 or 100 microg Abeta but resulted in comparable humoral immunity. Thus, Abeta vaccination is affected by the genetic background and amount of Abeta peptide used as immunogen. These data may help explain some differences observed in Abeta immunization studies in mice of various genetic backgrounds and aid in the design of Abeta vaccines. PMID- 15364460 TI - Surveillance of congenital rubella syndrome in Japan, 1978-2002: effect of revision of the immunization law. AB - Infection of rubella virus at the early stages of pregnancy in women who are not immune to rubella often induces congenital anomalies in infants, namely congenital rubella syndrome (CRS). This paper is the first comprehensive report of CRS cases in Japan, compiled from a questionnaire to major hospitals, reports to journals and academic meetings, and cases for virus/virus genome verification submitted to the National Institute of Infectious Diseases. CRS incidence in Japan was determined to be 0.2-8.1 cases/100,000 live births per year in epidemic years and 0.1-0.7 in non-epidemic years, respectively. In the last 4 years, the number of CRS cases remarkably decreased to one-three cases per year. This decrease is thought to be because the immunization law was revised in 1994 for changing the focus of rubella immunization from junior high school girls to infants of both sexes. PMID- 15364461 TI - Prevention of viral hepatitis in Italy. AB - The overall situation on viral hepatitis prevention and control in Italy was reviewed and evaluated at a Viral Hepatitis Prevention Board (VHPB) meeting in Catania, Sicily, on 7-8 November 2002. Several specific conclusions, drawn from the presentations and discussions, were considered to constitute an example of how to handle these issues in other European and industrialized countries. PMID- 15364462 TI - Double-blind randomized efficacy field trial of alum precipitated autoclaved Leishmania major vaccine mixed with BCG against canine visceral leishmaniasis in Meshkin-Shahr district, I.R. Iran. AB - This study was designed to evaluate the efficacy of a single dose of aluminum hydroxide (alum) precipitated Leishmania major (Alum-ALM) vaccine plus BCG against canine visceral leishmaniasis. Three hundred and forty-seven healthy dogs with no anti-Leishmania antibodies were double-blind randomly injected intradermally with either 0.1ml of Alum-ALM (200 microg protein) mixed with BCG (182 dogs) or injected with 0.1ml of normal saline (165 dogs). The results of 16 months follow-up showed that the vaccine was safe and well-tolerated. Strong seroconversion using DAT and ELISA techniques at 16 months post-vaccination was considered as an indication of Leishmania infection. The incidence rate was 3.7% (6/162) in vaccinated group and 12.0% (17/141) in control group using DAT technique. The efficacy of the vaccine was calculated to be 69.3%. PMID- 15364463 TI - Evidence supporting the inclusion of strains from each of the two co-circulating lineages of H3N8 equine influenza virus in vaccines. AB - Two lineages of antigenically distinct equine influenza A H3N8 subtype viruses, American and European, co-circulate. Experiments were conducted in ponies to investigate the protection induced by vaccines containing virus from one lineage against challenge infection with homologous or heterologous virus. Regression analysis showed that vaccinated ponies with average pre-challenge single radial haemolysis (SRH) antibody levels (i.e. 45-190mm2) had a higher probability of becoming infected if they were vaccinated with virus heterologous to the challenge strain than if they were vaccinated with homologous virus. Field studies in Thoroughbred racehorses also showed that SRH antibody levels of >/= 150mm2 induced by vaccines containing a European lineage strain are protective against infection with a virus from the same lineage, but that the same or higher antibody levels may not be protective against an American lineage virus. In conclusion, vaccines should contain virus strains representative of both H3N8 subtype lineages to maximise protection against infection. PMID- 15364464 TI - Waning immunity and subclinical measles infections in England. AB - Following the introduction of vaccination against measles, levels of clinical infection have dropped markedly. As we move further into the vaccine era, increasingly many individuals owe their measles immunity to vaccination and have had few (if any) exposures to wild virus. A number of recent reports suggest that vaccinated individuals with low levels of immunity may be at risk of subclinical measles infection. We explore the interplay between levels of infection and immunity over time using a mathematical model that simulates infection, waning and boosting of immunity. We focus particularly on the situation in England, where vaccination has been in place since 1966. Simulations of our model demonstrate a rise in the levels of subclinical measles infection over time, and a corresponding rise in clinical measles infections if vaccination levels are too low. We compare the impact of intervention strategies, and find that the rise in cases is most effectively reduced by 'catch-up' vaccination of children. In recent years, vaccination levels in England have dropped from above 90% in the 1990s to 84% in 2001/2002. We discuss the impact of declining vaccination levels on clinical and subclinical infections. PMID- 15364465 TI - Evidence that use of an inactivated equine herpesvirus vaccine induces serum cytotoxicity affecting the equine arteritis virus neutralisation test. AB - Several laboratories worldwide have recently experienced problems related to serum cytotoxicity with the equine arteritis virus (EAV) neutralisation test (VN) when using Office International des Epizooties (OIE) reference laboratory prescribed rabbit kidney (RK-13) indicator cells. Cytotoxicity can be mistaken for viral cytopathic effect and has led to increasing difficulties in test interpretation, consequently causing disruption to both equine breeding and disease surveillance. Results from experimental and field-derived data suggest that this serum cytotoxicity is associated with use of a tissue-culture-derived equine herpesvirus vaccine, probably manifested through a vaccine-induced anti cellular antibody response directed against RK-13 cells. Two alternative EAV VN methods were shown to significantly reduce the effects of cytotoxicity (from 73 to <5% prevalence) among vaccinated horses but did not completely eliminate the problem. Use of ELISA-based tests, which are not affected by serum cytotoxicity but which are not currently recognised as international standards, should be evaluated as a useful backup in screening equine sera for EAV VN antibodies. PMID- 15364466 TI - Attenuated Salmonella typhimurium htrA mutants cause fatal infections in mice deficient in NADPH oxidase and destroy NADPH oxidase-deficient macrophage monolayers. AB - Salmonella live vaccine strains harbouring mutations in htrA, a stress protein gene, display increased susceptibility to oxidative stress in vitro. This is believed to be connected to their reduced virulence, perhaps due to impaired survival inside phagocytes, although this has never been formally proven. We report that the in vitro phenotype of increased susceptibility to oxidative stress of Salmonella typhimurium htrA mutants newly prepared by transduction is rapidly lost on subculture, with the mutants becoming as resistant as the parent for reasons that remain unclear. However, despite this change, htrA mutants are still attenuated in normal mice. In contrast, they were found to be lethal for gene targeted gp91phox-/- mice deficient in NADPH oxidase, as was a S. typhimurium SPI-2 mutant known to be virulent in gp9lphox-/- mice. Infection with htrA mutants caused little damage to primary bone marrow macrophage cultures from normal mice; conversely, they caused extensive damage to macrophages from gp9lphox-/- mice, with more than 60% reduction in cell numbers 2.5h after being infected. The parental wild type strain similarly caused extensive damage to macrophages from both normal and gp9lphox-/- mice, whereas an aroA live vaccine strain had no effect on either normal or gp9lphox-/- macrophages. Taken collectively, the present results suggest that htrA is somehow involved in resistance to oxidative stress in vivo, with the avirulence of htrA mutants in mice being due to mechanisms which involve NADPH oxidase and suppression of bacterial growth within macrophages. PMID- 15364467 TI - Prevalence of Clostridium species and behaviour of Clostridium botulinum in gnocchi, a REPFED of italian origin. AB - Sales and consumption of refrigerated processed foods of extended durability (REPFEDs) have increased many-fold in Europe over the last 10 years. The safety and quality of these convenient ready-to-eat foods relies on a combination of mild heat treatment and refrigerated storage, sometimes in combination with other hurdles such as mild preservative factors. The major hazard to the microbiological safety of these foods is Clostridium botulinum. This paper reports on the prevalence and behaviour of proteolytic C. botulinum and non proteolytic C. botulinum in gnocchi, a potato-based REPFED of Italian origin. Attempts to isolate proteolytic C. botulinum and non-proteolytic C. botulinum from gnocchi and its ingredients were unsuccessful. Based on assessment of the adequacy of the methods used, it was estimated that for proteolytic C. botulinum there was < 25 spores/kg of gnocchi and < 70 spores/kg of ingredients. The total anaerobic microbial load of gnocchi and its ingredients was low, with an estimated 1 MPN/g in processed gnocchi. Most of the anaerobic flora was facultatively anaerobic. A few obligately anaerobic bacteria were isolated from gnocchi and its ingredients and belonged to different Clostridium species. The protection factor, number of decimal reductions in the probability of toxigenesis from a single spore, was determined for eight different gnocchi formulations by challenge test studies. For all gnocchi stored at 8 degrees C (as recommended by the manufacturer) or 12 degrees C (mild temperature abuse), growth and toxin production were not detected in 75 days. The protection factor was >4.2 for proteolytic C. botulinum, and >6.2 for non-proteolytic C. botulinum. When inoculated packs were stored at 20 degrees C (severe temperature abuse), toxin production in 75 days was prevented by the inclusion of 0.09% (w/w) sorbic acid (protection factors as above), however in the absence of sorbic acid the packs became toxic before the end of the intended shelf-life and the protection factors were lower. Providing sorbic acid (0.09% w/w) is included in the gnocchi, the safety margin would seem to be very large with respect to the foodborne botulism hazard. PMID- 15364468 TI - A selective differential medium for Enterobacter sakazakii, a preliminary study. AB - Enterobacter sakazakii can cause fatal invasive infection of neonates associated with the presence of this organism in powdered infant milk formula. A new chromogenic medium (Druggan-Forsythe-Iversen agar, DFI) is described for the selective detection of this emergent pathogen. The medium is based on the alpha glucosidase reaction which is detected using 5-bromo-4-chloro-3-indolyl-alpha,D glucopyranoside (XalphaGlc). Ent. sakazakii hydrolyses this substrate to an indigo pigment, producing blue-green colonies on this medium. DFI was compared with the current method of detection on violet red bile glucose agar (VRBGA) followed by pigment production on tryptone soy agar (TSA) after 48-72 h at 25 degrees C and subsequent biochemical profile determination using Biomerieux API20E. Ninety-five clinical and food strains of Ent. sakazakii were detected on the DFI chromogenic medium 2 days sooner than the alternative method. The characteristics of 148 strains representing 17 genera of non-Ent. sakazakii Enterobacteriaceae were compared using the two methods. Only 16/18 Escherichia vulneris strains, 2/3 strains of Pantoea spp. and 1/8 Citrobacter koseri strains gave false positive results on DFI agar. Eight alpha-glucosidase positive strains were identified as Pantoea using their API20E biochemical profile, but had higher percentage identification as Ent. sakazakii using ID32E. Therefore the DFI medium enables the detection of Ent. sakazakii within mixed cultures of Enterobacteriaceae, whereas the organism could be missed when using VRBGA since the latter is a general Enterobacteriaceae selective medium. In addition, the common use of API20E to check yellow pigmented colonies on TSA may lead to false negative results and consequently the acceptance of a batch of infant formula milk (IFM) that contains Ent. sakazakii. PMID- 15364469 TI - Evaluation of meat born lactic acid bacteria as protective cultures for the biopreservation of cooked meat products. AB - In this study, 91 strains, originating from meat products, were subjected to a step-by-step screening and characterisation to search for potential protective cultures to be used in the cooked cured meat industry. Strains were first tested on their homofermentative and psychrotrophic character and salt tolerance. Secondly, the antibacterial capacities towards Listeria monocytogenes, Leuconostoc mesenteroides, Leuconostoc carnosum and Brochotrix thermosphacta were determined in an agar spot test. In total, 38% of the tested strains were inhibitory towards all indicator strains. However, 91%, 88% and 74% of the strains could inhibit, respectively, L. monocytogenes, B. thermosphacta and Leuc. mesenteroides. Finally, 12 strains, with the highest antibacterial capacities, were evaluated on their competitive nature by comparing their growth rate, acidifying character and lactic acid production at 7 degrees C under anaerobic conditions in a liquid broth. All 12 strains, except for a bacteriocin producing Lactobacillus plantarum strain and the lactocin S producing Lactobacillus sakei 148, combined a fast growth rate with a deep and rapid acidification caused by the production of high levels of lactic acid. The 12 selected strains were then further investigated for their growth capacity on a model cooked ham product to establish whether the presence of these cultures on the ham did not negatively influence the sensory properties of the ham. All strains grew in 6 days at 7 degrees C from a level of 10(5)-10(6) to 10(7)-10(8) cfu/g and again the bacteriocin producing L. plantarum strain was the slowest growing strain. As the glucose level of the model cooked ham product was low (0.09+/-0.03%), growth of the putative protective cultures resulted in glucose depletion and a limited lactic acid production and accompanying pH decrease. Cooked ham inoculated with isolates 13E, 10A, 14A (all three identified as L. sakei subsp. carnosus by SDS PAGE) and with strains L. sakei 148 (LS5) and L. sakei subsp. carnosus SAGA 777 (LS8) were not rejected by the sensory panel at the 34th day of the vacuum packaged storage at 7 degrees C. Therefore, these strains could have potential for the use as protective culture in cooked meat products. PMID- 15364470 TI - Fast induction of nisin resistance in Streptococcus thermophilus INIA 463 during growth in milk. AB - Streptococcus thermophilus INIA 463 became nisin-resistant after exposure in skim milk to subminimal inhibitory concentrations of nisin (1-3 IU/ml) for less than 2 h. Addition of 20 IU/ml caused a 4 log unit decrease in S. thermophilus counts of a culture not exposed previously to nisin, whereas no decrease was observed in the culture exposed to nisin for 2 h. Transfer of immunity genes as responsible for nisin resistance was discarded. The presence of extracellular or intracellular specific nisin-degrading enzymes was not detected in the nisin resistant variant of S. thermophilus INIA 463. Nisin resistance was caused by the induction of a resistance mechanism. Transmission electron microscopy (TEM) revealed that the nisin-resistant variant of S. thermophilus INIA 463 had a thickened cell wall compared to the wild strain. Resistance to nisin was lost after one transfer (4 h growth) in nisin-free skim milk. PMID- 15364471 TI - Species identification by genotyping and determination of antibiotic resistance in Campylobacter jejuni and Campylobacter coli from humans and chickens in Sweden. AB - Campylobacter is today the most common cause of human bacterial enteritis in Sweden, as well as in most other industrialized countries. Common sources of infection are undercooked chicken meat, unpasteurized milk and contaminated drinking water. One aim with our present study was to identify the species Campylobacter jejuni and Campylobacter coli strains from humans and chickens using a polymerase chain reaction/restriction enzyme analysis (PCR/REA) method, as well as traditional hippurate hydrolysis test. Another aim was to investigate the antibiotic resistance pattern of the human domestic C. jejuni/C. coli isolates from infected patients and isolates from healthy Swedish chicken, as well as isolates from humans infected abroad. If discrimination between C. jejuni and C. coli was based on testing for hippurate hydrolysis, 95% of the human domestic strains and 88% of the chicken strains were identified as C. jejuni. Based on genotyping by PCR/REA, 100% of the human domestic strains and 98% of the chicken strains were attributed to C. jejuni. The E-test and disc diffusion methods were used for phenotypic antibiotic resistance studies. The two methods gave similar results. Most Swedish C. jejuni/C. coli isolates both from humans and chickens were sensitive to doxycycline and erythromycin, which are antibiotics used to treat human infection. Only 7% of the human domestic strains and 2% of the chicken strains were resistant to the quinolones tested. As a comparison, more than 94% of strains isolated from travelers to Asia and southern Europe showed antibiotic resistance to one or more drugs. PMID- 15364472 TI - Growth of Listeria monocytogenes as influenced by viscosity and water activity. AB - The effects of osmotic stress on Listeria monocytogenes growth parameters was examined in relation to the viscosity of the growth media. In low-viscosity systems, growth of L. monocytogenes in glucose-supplemented media was comparable to growth in sucrose-supplemented media. The relative lag time (RLT: the lag time divided by the generation time) responses were found to increase in the more restrictive water activity conditions. In high-viscosity systems containing polyvinylpyrrolidone (PVP), growth rate was reduced, whereas lag time showed no discernible modification. Osmotic stress in medium- and high-viscosity media supplemented with glucose resulted in approximately exponential increasing of the RLT values. Thus, the biological effects of osmotic stress on L. monocytogenes could be affected by the physical properties of the system, such as viscosity and diffusivity. PMID- 15364473 TI - Shiga toxin-producing Escherichia coli in healthy young beef steers from Argentina: prevalence and virulence properties. AB - Between July 1999 and December 2000, the prevalence of Shiga toxin-producing Escherichia coli (STEC) was established in 200 Argentine healthy young beef steers (14-16 months old) grown under local production systems with a feed grain period of 3-4 months, and the STEC strains isolated were examined in regard to their phenotypic and genotypic characteristics. Stool samples (n = 70) and rectal swabs (n = 130) were taken at the slaughterhouse level. By polymerase chain reaction (PCR), Shiga toxin (stx) gene sequences were detected in 69% of the samples. Eighty-six STEC strains were isolated from 39% of the animals. Serogroups identified, in order of frequency, were: O8 (16 strains), O113 (14), O103 (5), O91 (4), O171 (3), O174 (3), O25 (2), O112 (2), O145 (2), O2, O11, O104, O121, O128, O143, O146, O157. The most frequent serotype isolated was O8:H19 (12.9%). A total of 17 serotypes, including E. coli O157:H7 found in one animal (0.5%), have been previously associated with hemolytic uremic syndrome (HUS), bloody and non-bloody diarrhea in different countries, including Argentina. The prevalent genotype isolated was stx2 (51 of 86, 59.3%). Subtyping of stx2 variants showed the prevalence of stx2vh-b (25.6%) and stx2vh-a types (24.4%), and revealed the presence of an atypical stx2-v. Only 7.0% of STEC strains carried eae, and 33.7% harbored EHEC-hlyA gene. The full virulent genotype (stx/eae/EHEC-hlyA) was found to be present in 4 of the 86 (4.7%) STEC strains isolated. This research indicates that young steers from the main beef producing area of Argentina are an important reservoir of STEC strains; however, its importance as agents of human diseases in our country has still to be established. PMID- 15364474 TI - Tannase activity by lactic acid bacteria isolated from grape must and wine. AB - We examined a range of oenological lactic acid bacteria species and reference strains for their potential to degrade tannins. Bacterial tannase activity was checked by a spectrophotometric and a visual reading method. None of the strains belonging to the oenological species of the genus Lactobacillus, Leuconostoc, Oenococcus or Pediococcus were tannase producers, with the exception of Lactobacillus plantarum. All the L. plantarum strains analyzed were positive for tannase activity and their identities were reconfirmed by L. plantarum PCR specific assay or by sequencing the 16S rDNA. Tannase activity could be considered an important criterion for the selection of malolactic starter cultures since it might confer advantages in the winemaking process by reducing astringency and haze in wine. PMID- 15364475 TI - The sequence heterogenicities among 16S rRNA genes of Salmonella serovars and the effects on the specificity of the primers designed. AB - Previously, we have reported a 16S rDNA targeted polymerase chain reaction (PCR) method for the specific detection of Salmonella serovars [J. Appl. Bacteriol. 80 (1996) 659]. The target sites of its primers, i.e. 16SFI and 16SIII, according to the data in GenBank, were found mismatched to the corresponding sequences of some Salmonella serovars, such as those of S. Houten, S. Chingola, S. Bareilly, and S. Weltevreden. Accordingly, a PCR method using a nonspecific primer MINf combined with a primer modified from our 16SFI primer, i.e. the primer MINr, was developed and displayed better detection specificity [Int. J. Food Microbiol. 80 (2003) 67]. In this study, we show the sequence heterogenicity at the primer 16SFI targeting sites for some Salmonella serovars. Thus, the sequence used for designing of PCR primers might be just one of the several possible sequences. Such a situation may lead to the misjudgment on evaluation of the specificity of the primers if this was only based on the data in GenBank. Strains of the above described Salmonella serovars with target sequences from GenBank mismatched to the primer 16SF1 were reidentified and their PCR results were confirmed. Meanwhile, their 16SFI/16SIII primer annealing sites were sequenced and the sequences obtained were found completely and highly homologous to those of 16SFI and complementary to those of 16SIII primer, respectively. PMID- 15364476 TI - Posterior neocortical (visual cortex) lesions in the rat impair matching-to-place navigation in a swimming pool: a reevaluation of cortical contributions to spatial behavior using a new assessment of spatial versus non-spatial behavior. AB - In the face of contradictory findings on the role of visual cortex contributions to spatial behavior, the present study evaluated the ability of rats with primary visual cortex (Area 17) lesions to learn spatial problems in a swimming pool. Because the solution to any spatial learning problem consists of acquiring at least two primary elements of a task, task procedures and spatial learning, the study, in addition to assessing spatial ability on a place task, used two training/testing methods to identify the nature of the spatial impairment associated with visual cortex lesions. Non-spatial training consisted of learning to find a platform in the dark and spatial training consisted of a series of matching-to-place problems. The results confirmed that although rats with visual cortex lesions were impaired on place learning, the deficit was partially ameliorated by non-spatial training given following the lesion, and completely ameliorated by non-spatial training given before the lesion. Nevertheless, all visual cortex groups failed to show a quadrant preference on a probe trial and displayed a profound impairment in matching-to-place learning. This definitive demonstration that appropriate testing methods can reveal a failure in spatial behavior following visual cortex lesions is consistent with the idea that primary visual cortex is required in spatial navigation. PMID- 15364477 TI - Beta-amyloid (1-42)-induced learning and memory deficits in mice: involvement of oxidative burdens in the hippocampus and cerebral cortex. AB - We have demonstrated that oxidative stress is involved, at least in part, in beta amyloid protein (Abeta)-induced neurotoxicity in vivo [Eur. J. Neurosci. 1999;11:83-90; Neuroscience 2003;119:399-419]. However, mechanistic links between oxidative stress and memory loss in response to Abeta remain elusive. In the present study, we examined whether oxidative stress contributes to the memory deficits induced by intracerebroventricular injection of Abeta (1-42) in mice. Abeta (1-42)-induced memory impairments were observed, as measured by the water maze and passive avoidance tests, although these impairments were not found in Abeta (40-1)-treated mice. Treatment with antioxidant alpha-tocopherol significantly prevented memory impairment induced by Abeta (1-42). Increased activities of the cytosolic Cu,Zn-superoxide dismutase (Cu,Zn-SOD) and mitochondrial Mn-superoxide dismutase (Mn-SOD) were observed in the hippocampus and cerebral cortex of Abeta (1-42)-treated animals, as compared with Abeta (40 1)-treated mice. The induction of Cu,Zn-SOD was more pronounced than that of Mn SOD after Abeta (1-42) insult. However, the concomitant induction of glutathione peroxidase (GPX) in response to significant increases in SOD activity was not seen in animals treated with Abeta (1-42). Furthermore, glutathione reductase (GRX) activity was only increased at 2h after Abeta (1-42) injection. Production of malondialdehyde (lipid peroxidation) and protein carbonyl (protein oxidation) remained elevated at 10 days post-Abeta (1-42), but the antioxidant alpha tocopherol significantly prevented these oxidative stresses. Therefore, our results suggest that the oxidative stress contributes to the Abeta (1-42)-induced learning and memory deficits in mice. PMID- 15364478 TI - Regular voluntary exercise reduces anxiety-related behaviour and impulsiveness in mice. AB - We embarked on a study to delineate the behavioural changes in mice after 4 weeks of voluntary exercise. As an initial behavioural characterization, we exposed the control and exercising mice to a modified hole board and an open field test. As compared to control mice, exercising animals showed clear signs of increased behavioural inhibition (e.g. a longer latency to enter unprotected areas), suggesting increased anxiety in these animals. In addition, the exercising mice were reluctant to spend time in the open field's centre during the beginning of the 30-min open field test, but compensated for this at later times. Paradoxically, the exercising animals showed more rearings on the board of the modified hole board, indicating decreased anxiety. Thus, the behavioural inhibition seen in exercising mice is likely to represent decreased stress responsiveness at the behavioural level which can also be interpreted as reduced impulsiveness. To clarify whether voluntary exercise evolves in more or less anxiety-related behaviour, we exposed animals to the elevated plus-maze and the dark-light box, two selective tests for unconditioned anxiety. Clearly, compared to the control animals, exercising mice spent significantly more time on the open arm of the plus-maze and spent double the amount of time in the light compartment of the dark-light box. Taken together, we conclude that long-term voluntary exercise appears to result in decreased anxiety-related behaviour and impulsiveness. Thus, our observations fit into the concept that regular exercise strengthens endogenous stress coping mechanisms, thereby protecting the organism against the deleterious effects of stress. PMID- 15364479 TI - Protean behavior under barn-owl attack: voles alternate between freezing and fleeing and spiny mice flee in alternating patterns. AB - When attacking a spiny mouse in an experimental arena, a barn owl launched a few attacks from distant perches, made repetitive short-distance swoops in each attack and remained in the vicinity of the prey while chasing it. The spiny mouse fled in response, and typically oriented to face the owl whenever it stopped. When attacking a vole, the barn owl performed a greater number of attacks from distant perches, and left the vicinity of the prey after a few short-distance chases or capture attempts. Voles responded to these attacks in unspecific combinations of freezing and fleeing, and did not turn to face the owl when they stopped. Four conclusions are drawn from these encounters. First, two strategies characterized these predator-prey interactions; in one, both predator and prey continuously maintained awareness of each other's location; whereas in the other they continuously attempted to avoid the attention of the other. Second, responses of spiny mice and voles were a manifestation of protean behavior, with spiny mice fleeing in an alternating pattern and voles alternating between running and freezing. Third, locomotor response to owl attack comprised behavior that is an augmentation of normal behavior, with voles clinging to the walls and spiny mice running with frequent and irregular changes in direction. Fourth, the different defensive responses accord with the motor capacities and habitat of each rodent species. All in all, these results demonstrate the dynamic and multidimensional nature of predator-prey interactions. PMID- 15364480 TI - Prepulse inhibition of the acoustic startle reflex in pigs and its disruption by d-amphetamine. AB - Prepulse inhibition (PPI) of the startle reflex is an operational measure of sensorimotor gating. The dopamine receptor agonist-mediated disruption of PPI in rats is widely used as a model of the sensorimotor gating deficiencies demonstrated in schizophrenia patients. As a possible tool for validation of a pig model of psychosis, we wished to verify the existence of PPI in landrace pigs and investigate the potential disruption of PPI by d-amphetamine (AMPH) in these animals. PPI of the acoustic startle reflex and its potential disruption by AMPH were investigated using three doses 0.5-1.5mg/kg with a paradigm including two levels of prepulses (82 and 88dB) and a prepulse (PP) interval of 60 and 120ms. We found an average PPI of the startle reflex of 25.6% and both of the investigated PP intensities and PP intervals were equally effective in this PP inhibitive paradigm. AMPH significantly disrupted PPI and, in spite of only the 0.5mg/kg dose proved statistically significant, the results indicate this to be dose-related. We have demonstrated the phenomenon of PPI of the startle reflex in landrace pigs and its disruption by d-amphetamine. Studies of sensorimotor gating defects could be a valuable additional tool in assessing pig models of neuropsychiatric disorders. PMID- 15364481 TI - Have no fear, erythropoietin is here: erythropoietin protects fear conditioning performances after functional inactivation of the amygdala. AB - This study investigated the capacity of erythropoietin (EPO) to protect fear conditioning performances against functional inactivation of the amygdala. We infused an excitotoxic dose of glutamate in the lateral nucleus of the amygdala (LA) of adult rats in order to block the output projections to brainstem areas controlling the expression of conditioned fear responses. Subsequently, animals with excitotoxic lesions in the LA displayed altered short and long-term fear conditioned responses, but the integrity of their general emotional reactivity was preserved, as indicated by their open-field behavior. EPO infused immediately after glutamate succeeded to protect the conditioned fear performances of rats. This effect was reliably represented on both short, and long-term memory tests of conditioned fear. This and other studies have supported the potent neuroprotective activity of EPO, discriminable both morphologically, and behaviorally. PMID- 15364482 TI - Inertial properties of the arm are accurately predicted during motor imagery. AB - In the present study, using the mental chronometry paradigm, we examined the hypothesis that during motor imagery the brain uses a forward internal model of arm inertial properties to predict the motion of the arm in different dynamic states. Seven subjects performed overt and covert arm movements with one (motion around the shoulder joint) and two (motion around both the shoulder and elbow joints) degrees of freedom in the horizontal plane. Arm movements were executed under two loading conditions: without and with an added mass (4kg) attached to the subject's right wrist. Additionally, movements were performed in two different directions, condition which implies changes in the arm inertia due to the inertial anisotropy of the arm. Our analysis was focused on the timing features of overt and covert movements measured by means of an electronic stopwatch. Durations of right-direction arm movements (low inertial resistance) were smaller compared to durations of left-direction arm movements (high inertial resistance). Additionally, loading the arm with an added mass of 4kg significantly changed the dynamics of motion: movements were indeed more prolonged under loaded conditions. In both cases, the duration of simulated movements mirrored that of overtly executed movements. Therefore, neither the inertial anisotropy of the arm nor the addition of an external mass affected the timing correspondence between overt and covert movement execution. These findings suggest that the brain internally represents the inertial properties of the arm and makes use of it both for sensorimotor control and for the generation of motor images. PMID- 15364483 TI - Alterations in brain antioxidant status, protein oxidation and lipid peroxidation in response to different stress models. AB - The aim of this study was to investigate the effects of different stress models on copper, zinc-superoxide dismutase (Cu,Zn-SOD), catalase (CAT) and selenium dependent glutathione peroxidase (Se-GSH-Px) activities, and reduced glutathione (GSH), protein carbonyl (PC) and lipid peroxidation marker (conjugated diene (CD) and thiobarbituric acid-reactive substances (TBARS)) levels in brain of rats, and to determine the most effective stress model according to each parameter. Rats were divided into four groups as following: control group (C), immobilization stress group (IS), cold stress group (CS) and immobilization-cold stress group (ICS). All stress models increased brain Cu,Zn-SOD and CAT activities, PC, CD and TBARS levels, plasma corticosterone levels and decreased brain GSH concentrations. Se-GSH-Px activity was increased in CS and ICS groups. When all stress models were taken into consideration, the highest increases in Cu,Zn-SOD and Se-GSH-Px activities were found in CS group. The lowest GSH level was seen in IS group. The highest increases in PC and TBARS levels were found in ICS group. The highest increase of CD concentration was seen in IS and ICS groups. Our results suggest that different stress models have different degrees of influences on enzymatic and non-enzymatic antioxidant defense systems, protein oxidation and lipid peroxidation in the brain. PMID- 15364484 TI - Characterization of motor skill and instrumental learning time scales in a skilled reaching task in rat. AB - Successful motor skill learning requires repetitive training interrupted by rest periods. In humans, improvement occurs within and between training sessions reflecting fast and slow components of motor learning [Karni A, Meyer G, Rey Hipolito C, Jezzard P, Adams MM, Turner R, et al. The acquisition of skilled motor performance: fast and slow experience-driven changes in primary motor cortex. Proc Natl Acad Sci USA 1998;95:861-8]. Here, these components are characterized in male and female rats using a model of skilled forelimb reaching and are compared to time scales of instrumental learning. Twenty female and 14 male adult Long-Evans rats were pre-trained to operate a motorized door (via a sensor in the opposite cage wall) to access a food pellet by tongue. Latencies between pellet removal and door opening were recorded as measures of instrumental learning. After criterion performance was achieved, skilled forelimb reaching was requested by increasing the pellet-window distance to 1.5cm. Reaching success was recorded per trial. Mean latencies decreased exponentially over sessions and no improvement within-session was found. Skill learning over eight training sessions followed an exponential course in females and a sigmoid course in males. Females acquired the skill significantly faster than males starting at higher baseline levels (P < 0.001) but reaching similar plateaus. Within-session improvement was found during the sessions 1-3 in females and 1-4 in males. Performance at the end of session 1 was not carried over to session 2. Learning curves of individual animals were highly variable. These findings confirm in rat that motor skill learning has fast and slow components. No within-session improvement is seen in instrumental learning. PMID- 15364485 TI - Developmentally stable sex-dependent modulation of turning asymmetry by neonatal novelty exposure. AB - In rats, early life stimulation can enhance learning and memory and induce parallel changes in brain asymmetry. Despite persistent interest in human brain asymmetry, relatively little is known in animal models about developmental stability of early-experience effects on asymmetry and how early-experience may affect males and females differently in asymmetry measures across developmental stages. We exposed male and female neonatal rats to a novel cage for 3min per day during the first 3 weeks of life and measured spontaneous turning behavior at juvenility (7 weeks of age) and adulthood (7 months of age). We found that (1) the effects of such neonatal novelty exposure on turning bias are developmentally stable, and (2) neonatal novelty exposure differentially modulates turning bias in males and females. We briefly discuss implications of these findings in terms of the role of brain asymmetry in modulating cognitive and emotional development. PMID- 15364486 TI - Chronic ultra-mild stress improves locomotor performance of B6D2F1 mice in a motor risk situation. AB - Chronic low grade stress predispose to psychopathological disorders. We consistently showed that chronic ultra-mild stress (CUMS) applied to B6D2F1 female mice induced behavioral disinhibition in several conflict exploration models. Insufficient reactivity to conflicts may be maladaptive and lead to inappropriate appreciation of potential risks and impaired ability to cope with those. Therefore, the purpose of the study was to assess the effect of a CUMS procedure on the behavior of mice in a motor risk situation based on multisensory conflict. Following three weeks exposure to various mild stressors, stressed and control B6D2F1 mice were tested on the rotating beam to determine how CUMS exposure affected balance control, posture stability and locomotor performance in response to a sensory-motor challenge. Detailed behavioral analysis included several parameters, both postural (height of the trunk, tail angle, number of imbalances, falls and head movements) and kinetic (mean velocity on the beam, distance covered with large and small movements, plus time spent in no-motion episodes). Comparisons between control and stressed mice showed that CUMS exposure increased mean velocity and improved locomotor performance in the learning task. In addition, sensitivity to sensory conflict seemed to be reduced in stressed mice, which displayed fewer behavioral adjustments to the increasing difficulty of the test compared to control mice. The results are discussed in terms of the possible influence of disturbances in behavioral and attentional inhibitory processes following CUMS exposure. Whether longer periods of CUMS exposure would shift the performance on the RTB from improvement to deterioration remain to be established. PMID- 15364487 TI - Effects of the 5-HT7 receptor antagonists SB-269970 and DR 4004 in autoshaping Pavlovian/instrumental learning task. AB - There is an important debate regarding the functional role of the 5-HT(1A) and 5 HT(7) receptor in memory systems. Hence, the objective of this paper is to investigate the function of serotonin (5-hydroxytryptamine, 5-HT) in memory consolidation, utilising an autoshaping Pavlovian/instrumental learning test. Specific antagonists at 5-HT(1A) (WAY 100635) and 5-HT(7) (SB-269970 or DR 4004) receptors administered i.p. or s.c.) after training, significantly decreased the improvement of performance produced by the 5-HT(1A/7) agonist 8-OH-DPAT to levels lower than controls'. These same antagonists attenuated the decreased level of performance produced by mCPP, although they decrease the performance levels after p-chloroamphetamine (PCA) lesion of the 5-HT system, which has no effect on its own on the conditioned response. Moreover, SB-269970 or DR 4004 reversed amnesia induced by scopolamine and dizocilpine. These data confirm a role for 5-HT(1A) and 5-HT(7) receptors in memory formation and support the hypothesis that serotonergic, cholinergic, and glutamatergic systems interact in cognitively impaired animals. These findings support a potential role for both 5-HT(1A) and 5 HT(7) receptors in the pathophysiology and/or treatment of schizophrenia, cognitive deficits and the mechanism of action of atypical antipsychotic drugs. PMID- 15364488 TI - A behavioural characterisation of the FVB/N mouse strain. AB - The use of transgenic models in scientific research has made an enormous contribution to our understanding of the causes and symptoms of many diseases, including neurodegenerative conditions such as Alzheimer's Disease (AD) and Parkinson's Disease (PD). In the creation of transgenic models of neurodegenerative disease, effects of the background strain of the animal on the resulting genotype must be taken into consideration. This is particularly true for behavioural studies in which the background strain of the mouse may mask the phenotype of the genetic manipulation. Here, the behaviour of two mouse strains used in transgenic models, FVB/N and C57BL6/J, were compared. Studies of circadian wheel activity, cognition and aggression revealed considerable phenotypic differences between strains. These data also indicate that the FVB/N strain is not appropriate as a background strain in the behavioural assessment of transgenic mouse models. PMID- 15364489 TI - Susceptibility to subchronic unpredictable stress is related to individual reactivity to threat stimuli in mice. AB - As in many complex behavioral responses, inter-individual variability can be observed in the responses to a chronic mild stress. While some subjects exhibit more resilient behaviours, others appear more susceptible to stress. This study hypothesizes that this variability relies on the individual appraisal of the stressful event. To study this assumption, mice were first subjected to a conditioned task occurring in a circular arena. In this task, a mild air-puff (i.e. stressor) in a given quadrant of the arena was coupled with the presence or the absence of a light in the same quadrant. Half of mice were then submitted to a 15-day subchronic stress consisting in various environmental and social mild stressors randomly applied two or three times a day. At the end of this procedure, the occurrence of depressive-like behaviours in stressed mice was assessed using measures of the stress regime (i.e. physical state, choice test, grooming test). The physical state assessed the physical appearance of mice. The grooming test consisted in measuring the time spent in grooming after mice were sprayed upon with a viscous solution. The choice test consisted in measuring the time spent in an uncomfortable place (i.e. whose floor was covered with damp sawdust) versus a more comfortable one (i.e. with dry sawdust) to evaluate the reactivity to a negative stimulus previously encountered during the subchronic stress. Multiple regression analyses revealed a relationship between attention toward salient stressful stimuli in the conditioned task and susceptibility to the subchronic stress procedure. These results are discussed regarding their relevance for the understanding of aetiologies of depressive illnesses. PMID- 15364490 TI - Medial preoptic area delta-opioid receptors inhibit lordosis. AB - Endogenous opioid peptides that activate the delta-opioid receptor (DOR) are thought to facilitate female receptive behavior. This facilitation of lordosis has been demonstrated by intracerebroventricular infusions and injection of DOR active ligands into the ventromedial hypothalamic nucleus, an area with robust DOR binding. However, DOR binding is distributed throughout the hypothalamus, and the role of DOR in other areas of the hypothalamus has not been examined. In the current study, we demonstrated DOR immunoreactivity in the medial preoptic area (MPO), in particular medial preoptic nucleus (MPN) of the preoptic area. DOR immunoreactive processes were sparsely distributed in the medial and lateral parts of the MPN. Larger DOR immunoreactive fibers were localized in the ventrolateral aspect of the lateral MPN. The MPN is involved in the modulation of female sexual receptivity and the distribution of DOR in this area suggested to us that DOR may regulate lordosis. Ovariectomized rats with unilateral cannulae aimed at the MPN were given 5microg 17beta-estradiol benzoate (EB), once every 4 days and tested for lordosis. [D-Pen(2), D-Pen(5)]-enkephalin (DPDPE), a DOR agonist, microinfused into the MPO, 52-54h after EB-priming, inhibited lordosis when compared with the aCSF (vehicle) control (P <== 0.05). The inhibitory effects of DPDPE were reversed by microinjection of naltrindole, a DOR antagonist (P <== 0.05). Interestingly, the DOR inhibition of lordosis is similar to the micro-opioid receptor inhibition of lordosis in the MPN. These results indicate that DOR in the MPO, particularly in the MPNm, plays an important role in the regulation of lordosis. PMID- 15364491 TI - Vocal production in different social contexts relates to variation in immediate early gene immunoreactivity within and outside of the song control system. AB - In songbirds, a major function of song during the breeding season is mate attraction, and song in this context can be highly sexually motivated. Vocal learning, perception, and production are regulated by the song control system, but there is no evidence that this system participates in the motivation to sing. Instead, brain regions involved in sexual motivation and arousal, including the medial preoptic nucleus (POM), bed nucleus of the stria terminalis (BST), nucleus taeniae (Tn), and area ventralis of Tsai (AVT) might regulate the motivation to sing, at least in a sexual context. The role of these nuclei and song control nuclei (area X and HVC) in vocal production within a breeding context, and other courtship behaviors, was investigated using immunocytochemistry for protein products of immediate early genes (IEGs), ZENK and c-fos (Fos), in flocks of male house sparrows (Passer domesticus) presented with females. Compared to vocalizations from other perches, vocal behavior from a nest box is more likely directed toward females, and sexually motivated. The numbers of ZENK and Fos labeled cells within rostral, but not caudal POM related positively only to vocalizations produced from a nest box. In contrast, the number of ZENK-labeled cells within area X related negatively to vocalizations from a nest box. Additionally, numbers of IEG-labeled cells within rPOM, Tn and AVT related positively to mount attempts. The results support the hypothesis that the POM interacts with the song control system to regulate sexually motivated vocal expression, and are consistent with work indicating that (a) rostral and caudal POM play distinct roles in sexual behavior, and (b) involvement of area X in song is context specific. PMID- 15364492 TI - Food restriction and leptin impact brain reward circuitry in lean and obese Zucker rats. AB - The rewarding effect produced by electrically stimulating certain sites in the lateral hypothalamus (LH) can be potentiated by food restriction and body weight loss in lean rats. Central leptin and insulin administration can suppress the rewarding impact of the stimulation. To determine whether there are additional peripheral signals that mediate the effect of weight loss on brain reward circuitry, we assessed changes in LH-self-stimulation following food restriction in the obese Zucker rat which develops resistance to circulating leptin and insulin. In addition, we examined the impact of acute food deprivation and leptin administration on LH self-stimulation in lean and obese Zucker rats. The number of brain stimulation rewards earned was measured over a range of LH stimulation frequencies that drove reward rates from zero to asymptotic levels. Restriction reduced frequency thresholds in a subset of lean and obese rats, whereas BSR was unaltered by acute food deprivation. Despite impairment in leptin signaling, intraventricular leptin (4 microg) increased thresholds in most lean and obese rats in which the rewarding effect was sensitive to restriction. These results show that brain reward circuitry in the obese Zucker rat is sensitive to weight loss and leptin. PMID- 15364493 TI - Behavioral audiometry: protocols for measuring hearing thresholds in babies aged 4-18 months. AB - OBJECTIVE: This paper provides the first report in English of original behavioral audiometry protocols for measuring hearing thresholds in very young children, including the multiply handicapped. METHODS: Based on reactions to one or two well-calibrated acoustic stimulations delivered in the sound field, the protocol first involves the use of a vibrator to measure hearing levels by bone conduction. This measurement technique, which is not affected by middle ear infections, is the key diagnostic step. Moreover, in profoundly hearing loss children, it triggers reactions through vibratory stimulation and sets the scene for the conditioning of responses. Next, hearing levels are assessed by air conduction with the aid of headphones, in order to measure hearing levels in each ear as early as possible. A unique set-up is used to facilitate the emergence of reliable "surprise reactions", which may be interpreted by a sole examiner. Classical visual reinforcement is replaced by a highly interactive, dynamic and playful exchange between child and examiner, which gives meaning to the perception of stimuli and heralds the learning of hearing. RESULTS: The results concern 105 babies suffering from bilateral sensorineural hearing loss and aged 4 18 months at the first behavioral test. Group 1 comprised 91 babies with no other handicap, in whom full bilateral air conduction was obtained in 82.4% before 12 months and in 98.9% before 18 months. In this group, air conduction in each ear was obtained in 47.0% before 12 months and in 70.3% before 18 months. In Group 2, which included 14 multiply handicapped babies, full bilateral air conduction was obtained in 37.5% before 12 months and in 78.6% before 18 months. Air conduction in both ears was obtained in 28.6% before 18 months. CONCLUSION: The protocols described make it possible, in a minimum number of sessions, to measure hearing thresholds early over the whole range of hearing frequencies, even in multiply handicapped babies and those suffering from developmental retardation. PMID- 15364494 TI - Sensorineural hearing loss in Jewish children born in Jerusalem. AB - OBJECTIVE: Many factors, hereditary and environmental, may cause deafness. The aim of the present study was to analyze data on the etiology of bilateral sensorineural hearing impairment in children born in the Jerusalem area during 1978-1991, and to compare the results to those of a previous survey (1968-1977) in the same area. METHODS: The study included 150 Jewish children (139 families) with hearing loss, born during 1978-1991. Information was obtained on prenatal, perinatal and postnatal events, history of hearing loss in the family, the parents' communities and consanguinity. Children with a sensorineural hearing impairment of 56 dB HL or greater in the better ear, within the frequency range of 0.5-4 kHz were included in the study. The hearing loss was classified as moderate-severe (56-70 dB HL), severe (71-90 dB HL) and profound (91 dB HL or more) in the better ear. Mutations in the coding sequence of the connexin 26 (C x 26) and the connexin 30 genes were examined in some of the families. RESULTS: The hearing impairment was hereditary in 66 (44%) of the children, environmental in 31 (21%) and four children (3%) had multiple malformations. The cause was unknown in 49 (33%) children. Sixty-two families were of European origin (Ashkenazim) and 62 of Afro-Asian origin (Sephardim). Consanguinity was in 7% of the families. Mutations in connexin 26 and the deletion in connexin 30 were diagnosed in 9/18 families tested. The incidence of hearing loss decreased from 1.28 per thousand during 1968-1977 to 1.06 per thousand during 1978-1991. The rate of environmental causes decreased over the years together with an increase in the rate of unknown causes. The rate of hearing loss among Sephardim decreased significantly and increased among Ashkenazim. CONCLUSIONS: The rate of hearing impairment in Israel is as that found in other countries, as was the distribution of the causes of deafness. The decrease in the rates of hearing impairment among the Sephardim may be due to a continuing decrease in consanguineous marriages among Sephardim. It is expected that the group of unknown causes will become smaller in future with the availability of more molecular genetic tests. PMID- 15364495 TI - Myxomatous lesions of the maxilla in children: a case series and review of management. AB - OBJECTIVE: A paucity of data exists in the literature regarding the pediatric bony myxoma. Controversy exists regarding both its histopathologic origins and surgical management. The purpose of this study was to report on our experience in managing these patients, including diagnostic work-up, histopathologic findings and an evaluation of surgical treatment outcomes. METHODS: A retrospective review was undertaken of all consecutive cases of pediatric bony myxoma treated between 1988 and 2001. Tumors were analyzed in terms of clinical data, imaging studies, histopathology and surgical treatment. A selective review of the literature regarding management of the bony myxoma is also presented. RESULTS: Five lesions were identified, all of which occurred in the maxillary bone. Histopathologically our cases did not show a strong association with odontogenic epithelium. All cases were treated by surgical resection with conservative margins. No recurrences were documented over a mean follow-up period of 8.5 years. CONCLUSION: The bony myxoma is a rare lesion in the pediatric population, and initial clinical or radiological misdiagnoses are not uncommon. The presupposed odontogenic origin of these tumors may need reassessment. Conservative resection with narrow margins is a safe and effective therapy for myxomas of the pediatric maxilla. PMID- 15364496 TI - Bilateral cochlear implants: a way to optimize auditory perception abilities in deaf children? AB - OBJECTIVES: The Wurzburg bilateral cochlear implant (CI) program was started with the aim to improve the patients' communicative abilities in complex listening situations. In this study, the auditory skills of children using bilateral cochlear implants were evaluated. STUDY DESIGN AND SETTINGS: Qualitative data based on free observations in the rehabilitation setup were collected in 39 bilaterally implanted children. A speech discrimination in noise test was performed in 18 of these children; lists of bisyllabic words were presented in noise at a signal to noise ratio (SNR) of +15 dB. RESULTS: Qualitative and quantitative data show clearly that bilateral CI improves the children's communicative behaviour, especially in complex listening situations. Children examined with the speech in noise test scored significantly better under the bilateral condition compared to the unilateral condition. Integration of the second implanted side and use of binaural information was observed to be easier and faster in children with a short time lag between both implants. CONCLUSIONS: To be able to obtain optimal benefit from bilateral cochlear implants, an intensive rehabilitation program is necessary. The important aspects of such a program are creating realistic expectations in older children before implantation; performing the first processor fitting of the second side with the first side switched on; and separate intensive training with the new system in order to balance out the hearing competence of the second CI with that of the first. PMID- 15364497 TI - Effect of methylphenidate on auditory event related potential in boys with attention deficit hyperactivity disorder. AB - OBJECTIVE: Event related brain potentials (ERPs) is a non-invasive technique giving knowledge about neural activity associated with sensory and cognitive information processing. The aims of the present study were to investigate amplitude and latency of P100, N200, and P300 in parietal and frontal areas in children with attention deficit hyperactivity disorder (ADHD), and in healthy children, and to determine the effect of methylphenidate (MPH) on these ERPs indices in ADHD group. METHODS: ERP indices, latencies of parietal P3 (PP3L), P1 (PP1L), N2 (PN2L), and frontal P1 (FP1L), N2 (FN2L), P3 (FP3L), and amplitudes of parietal P3 (PP3A), P1 (PP1A), N2 (PN2A), and frontal P1 (FP1A), N2 (FN2A), and P3 (FP3A), using an auditory oddball paradigm were recorded before and under MPH treatment in boys with ADHD, and in 23 healthy children. RESULTS: Before MPH treatment, PP3L was significantly longer and PP3A, PN2A, FN2A, and FP3A smaller in children with ADHD compared to healthy children (all P values < .05). No significant difference was found in PP1L, PP1A, PN2L, FP1L, FP1A, FN2L, and FP3L between ADHD and control group (all P values > .05). MPH treatment resulted in a significant decrease in PP3L, PN2L, and FP3L, and increase in PP3A, PP1A, and FP3A (all P values < .05). There was no significant difference in PP1L, PN2A, FP1L, FP1A, FN2L, and FN2A between before MPH and under MPH treatment in ADHD subjects (all P values > .05). Under MPH treatment, PP3L, PP3A, PP1L, PP1A, PN2L, FP1L, FP1A, FN2L, FP3L, and FP3A were not significantly different between children with ADHD and healthy controls (all P values > .05). However, PN2A and FN2A were significantly smaller in ADHD subjects compared to controls (both P values < .05). CONCLUSION: This study provides indirect evidence that ADHD subjects are associated with abnormalities in signal detection (inattention) and discrimination, and information processing. In addition, present study has shown that except FN2A and PN2A, MPH normalizes ERP indices, which suggested that MPH may be effective on impaired information processing in ADHD, but not on the receiving information. PMID- 15364498 TI - Thyroid surgery in children and adolescents. AB - Authors present 114 children and adolescents of the total 3865 patients that underwent surgery on the Department of Otorhinolaryngology and Head and Neck Surgery, 1st Faculty of Medicine, Charles University, Prague, Department of Otorhinolaryngology of IPVZ, Prague because of a thyroid disease in years 1991 2000. The male/female ratio in the group was 1:6.5, 16 (13.9%) males and 99 (86.1%) females. For non-malignant disease, the male/female ratio was 1:6.5 and for malignant disease, it was 1:5.4. Thyroid surgery was indicated for non malignant disease in 82 (71.9%) cases--11 males and 71 females. Most frequently it was indicated because of Graves-Basedow and Hashimoto toxicosis--in 39 children (34.2%), in 5 children (4.4%) for toxic adenoma. Because of the polynodular goiter surgery was indicated in 13 cases (11.4%), in 22 cases (19.3%) for mononodular goiter. Three girls (2.6%) were recommended for surgery because of Hashimoto thyreoiditis and for mechanic syndrome in diffuse goiter one girl (0.9%). Malignant tumor was found in 32 children (28.1%)--5 males and 27 females. Most frequent histological type was papillary cancer--in 25 cases (21.9%). Follicular cancer was diagnosed in four cases (3.5%) and medullar cancer in three cases (2.6%). TNM classification in children and adults is summarized in Table 1. Surgery was carried out in two children with distant metastases (lung localization). In both children the pre-disease stage lasted more than 12 months. The first manifestation of the disease was a finding of enlarged and palpable nodes on the neck. The authors summarize indications for surgery of thyroid diseases in children and adolescents. A treatment of the thyroid gland is a teamwork. Indication for surgery is carried out by a pediatric endocrinologist in cooperation with a surgeon specialized in thyroid surgery. The possible identification of the recurrent laryngeal nerve by a surgery microscope is an unquestionable advance. Children and adults should be centralized into health care centers capable not only of surgery, but also of endocrinology care. In children with a malignant disease there is a need for cooperation with a pediatric oncologist and a following oncological treatment on a nuclear medicine department. The authors summarize the information about today's thyroid surgery care possibilities for children and adolescents. They point out the differences in indications and the decision about extent of surgery in children and adolescents. PMID- 15364499 TI - Intrinsic and extrinsic forces determine the distortion of the split cricoid ring. AB - OBJECTIVE: An anterior cricoid split (ACS) causes an immediate distortion of the cricoid cartilage resulting in an anterior gap due to retraction of the cut ends. The objective of this animal study is to investigate: (1) to what extent the distortion after ACS is influenced by non-cartilaginous structures like tunica elastica, membranes, ligaments and muscles, which are connected to the cricoid; (2) how distortion is changing with further development; (3) in what way the distortion is affected by scoring of the internal surface of the cricoid; and (4) whether an immediate or late injury-induced distortion is related to age. METHODS: Surgical interventions were performed in 20 young (8 weeks of age, 1300 1600 g) and 5 adult (28 weeks of age, 3500-4000 g) New Zealand White rabbits. The immediate effects were measured, and then the animals were followed for 20 weeks to study the long-term effects of the various procedures. RESULTS: (1) The gap, immediately following an ACS, increased after additional transection of the cricothyroid ligament and the cricotracheal membrane, and even more when the cricovocal membrane was elevated from the inner surface of the cricoid arch. (2) The degree of distortion after various interventions in young animals appeared to increase substantially during further growth. (3) When the above-mentioned successive surgical steps were combined with scoring of the internal surface of the cricoid arch, a marked malformation of the split cricoid did develop with warping of the cut ends in lateral direction and a latero-cephalic rotation, the latter due to the action of the cricothyroid muscles. (4) The immediate distortion appeared to be similar in young and adult animals. During a follow-up of 20 weeks, a progressive distortion of the split cricoid ring was observed in the young growing rabbits. In adult animals, no significant progression of the distortion was found. CONCLUSIONS: The immediate and long-term distortion of the split cricoid is determined by the release of intrinsic forces of the cartilage, and extrinsic forces from non-cartilaginous structures like ligaments, muscles, membranes and tunica elastica. PMID- 15364500 TI - Cochlear implant positioning in children: a survey of patient satisfaction. AB - INTRODUCTION: The past decade has been witness to significant diminution in incision size and placement for pediatric cochlear implantation at our institution. Furthermore, the orientation of the long axis of the implant relative to the skull base plane has generally evolved from a retro-verted to ante-verted direction to accommodate the ear level receiver stimulator and the new minimal access technique. OBJECTIVE: To characterize the cosmetic, functional and clinical implications of this evolution in cochlear implant positioning by means of a patient satisfaction survey. METHODS: A survey of patient satisfaction was administered to 100 cochlear implant recipients or their parents. The survey elicits the patient's level of satisfaction with respect to implant position and functional limitations. In addition, the angle formed by the long axis of the implant and the skull base plane (implant angle) was determined from lateral skull radiographs to detect the temporal evolution in implant positioning. RESULTS: Analysis of the radiographs revealed an implant angle of 65.4-125 degrees in patients implanted prior to 1997, an implant angle of 56.3-87.5 degrees in patients implanted from 1997 to 2001, and an implant angle of 35.1 78.1 degrees after 2002. In spite of this evolution in positioning, the patient satisfaction survey did not reveal any change in incidence of cosmetic, functional or clinical problems. DISCUSSION: The significant changes in incision and implant positioning represent a technical evolution in pediatric cochlear implant surgery. The fact that patients have been consistently satisfied over time suggests that the cosmetic, functional and clinical implications are minimal. PMID- 15364501 TI - Aural irrigation using the OtoClear Safe Irrigation System in children. AB - OBJECTIVE: To evaluate the safety and efficacy of the OtoClear Safe Irrigation System for removing cerumen from the external auditory canal in children. METHODS: Eligible subjects were 6 months-17 years of age with cerumen obstructing > or = 50% of the tympanic membrane (TM) from view (by otoscopy). Pneumatic otoscopy, tympanometry, and audiometry were performed followed by cleansing of the affected ear canal(s) with the OtoClear Safe Irrigation System and warm tap water. Otoscopy was performed after each wash of the canal. A curette or small alligator forceps was used to remove remaining cerumen if necessary. Tympanometry and audiometry were repeated after all procedures were completed. Telephone contact was made 1 week later regarding symptoms of acute otitis externa or any other problems. RESULTS: Eighteen children (28 ears) ages 1-10/12 to 11-2/12 years were entered. Four had previously had tympanostomy tubes. At entry, there was no visible TM in 19 ears, 5-10% visible TM in 5 ears, 20% in 1 ear, and 30 40% of the TM in 3 ears. The number of washes needed per ear was: 1 wash--16 ears, 2 washes--8 ears, 3 washes--1 ear, 4 washes--2 ears; washing was stopped in 1 ear because of pain. After irrigation, a curette or forceps was used in 6 ears. Following the procedures, > or = 95% of the TM was visible in 24 ears, and > or = 80% was visible in all ears. Six ears (4 children) with flat tympanograms at entry became normal after irrigation. On audiometry, a conductive loss in 2 ears at entry resolved after irrigation. The mean change in pure tone average (PTA) was -2.9 dB. Three subjects were noted to have hearing losses >5 dB at some frequencies which on review by audiologists were deemed non-significant. No perforations of the TM occurred. There were no reports of otitis externa or any other adverse events occurring after leaving the clinic. CONCLUSION: We found the OtoClear Safe Irrigation System to be safe and effective in our small sample of children. It was well tolerated in most and provided a non-traumatic method for the removal of obstructing cerumen. PMID- 15364502 TI - Myringotomy and tympanostomy tube placement in children with sickle cell disease. AB - OBJECTIVE: To assess the event rate of myringotomy and tube placement (M&T) in the pediatric patient population with sickle cell disease (SCD). METHODS: Four hundred and forty-nine children with confirmed SCD have been followed over a period of 11.5 years at two hospital-based pediatric hematology and otolaryngology offices, and three tertiary care hospitals. Children with SCD who had undergone M&T were identified via computer search of International Classification of Diseases codes by the medical records departments of the three hospitals, and from two databases of the hematology offices. The inpatient and outpatient medical records of all children identified were reviewed. RESULTS: For the 449 patients, mean duration of SCD follow-up was 6.13 +/- 3.36 years. Of these, eight patients (four boys, four girls, mean age 9 +/- 3.5 years; four patients had hemoglobin SC disease, and four patients had sickle cell anemia type SS) underwent M&T. Two children met criteria for severe SCD. The event rate for M&T insertion was 0.29/100 person-years, 95% CI (0.15, 0.58). CONCLUSIONS: The event rate for M&T in children with SCD, compared to a historical control group, is lower than that of the general population. Type and severity of SCD were not predictive of the need for tube insertion. Children with sickle cell disease do not have an increased rate of M&T insertion. PMID- 15364503 TI - Causes of severe to profound deafness in Omani paediatric population. AB - The study includes a retrospective analysis of causes of deafness in 1400 Omani children who were detected to be suffering from severe to profound levels of hearing impairment. A standard form was used to collect various details from the otolaryngologists all over the country. The time period included is from 1986 to 2000. The rate of consanguineous marriage in the parents of the affected children was 70%, and 67.8% of these children had a sibling suffering from hearing impairment. There has been a rapid evolution of the antenatal, perinatal and neonatal care delivery in Oman during last couple of decades. The Omani paediatric population has been well covered (>90%) under the vaccination protocols including rubella. It has helped in reducing the share of infectious diseases towards the hearing impairment. The study is an attempt to identify various causes of the identifiable causes of severe to profound hearing impairment in Omani paediatric population which is the first attempt of this kind in this country. Relevant literature has been reviewed. PMID- 15364504 TI - Diagnostic accuracy and the observation option in acute otitis media: the Capital Region Otitis Project. AB - OBJECTIVE: Studies of acute otitis media indicate that deferring antibiotics in selected patients is a valid option. This study sought to determine the effects of a multifaceted educational intervention on disease management and treatment attitudes in practitioners caring for acute otitis media (AOM). METHODS: An expert committee composed of clinical and public health practitioners was convened which reviewed current evidence-based literature on diagnosis and treatment of AOM. A survey instrument to measure attitudes was fashioned and administered to 150 invited practitioners before and after a daylong educational intervention. The intervention was composed of two diagnostic otoscopy workshops; a lecture on an antibiotic-centered approach to treatment, followed by a lecture on the Dutch "observation without antibiotic option" (OWAO), which in 1998 involved withholding antibiotic prescription for selected patients over the age of 2 for up to 72 h; and a concluding "Miller Interactive Process"--small group sessions on five key clinical management questions. RESULTS: Significant short term changes occurred in practitioners' attitudes in importance of ear pulling as a diagnostic symptom (P = 0.034) and clinical findings in the tympanic membrane for the diagnosis of AOM (P = 0.006). Significant changes towards increased comfort of practitioners' management attitudes were found in managing selected patients without antibiotics (P = 0.057), using shortened courses of antibiotics (P = 0.001), and using high dose amoxicillin (80 mg/kg/day) as a second line drug (P = 0.001). CONCLUSIONS: Practitioners' attitudes about managing AOM can be modified in the short-term with an educational intervention combining didactic, hands-on, and interactive learning that incorporates evidence-based medicine. PMID- 15364505 TI - Inspiratory muscle strength training with behavioral therapy in a case of a rower with presumed exercise-induced paradoxical vocal-fold dysfunction. AB - Paradoxical vocal fold dysfunction (PVFD) with high effort exercise can result in disruptions to ventilation, dyspnea, inspiratory stridor, elevated heart rate, and syncope. This single subject study experimentally tested an inspiratory muscle strength training (IMST) program with behavioral therapy on a 15-year-old male crew member. Outcome variables were maximum inspiratory pressure (MIP), and dyspnea ratings. Following 5 weeks of IMST, MIP increased by 93% from baseline function while dyspnea ratings substantially decreased. Outcome included successful competition with his high-school crew team, a task he was previously unable to complete. Discussion focuses on IMST combined with traditional approaches of voice therapy for treating PVFD. PMID- 15364506 TI - Alveolar soft part sarcoma: report of a case occurring in the sinonasal region. AB - A primary alveolar soft part sarcoma arising in the nasal-paranasal sinuses region is quite rare. In this report, a 14-year-old boy with typical features of alveolar soft part sarcoma of the nasal and paranasal sinuses is presented. Treatment was surgical excision of the mass combined with radiotherapy. There has been no recurrence or metastasis as of 2 years after treatment. PMID- 15364507 TI - Solid aneurysmal bone cyst of the mandible. AB - Aneurysmal bone cyst is a rare, rapidly expanding, locally destructive, and often misdiagnosed lesion. It accounts for about 1-2% of primary biopsied bone tumours. About 60-70 cases have been reported in the jaws; particularly the molar regions. Eighty percent of patients are under 20 years of age. Aneurysmal bone cyst exists as a primary or secondary lesion. It may be conventional (95%) or solid (5%). The solid variant is more difficult to recognize. The practical importance of aneurysmal bone cyst lies in the fact that it must be differentiated from malignant tumours: mainly with giant cell tumours and teleangiectatic osteosarcoma. PMID- 15364508 TI - Auricular malignant melanoma of early childhood: the role of sentinel node biopsy. AB - Pediatric melanoma is a rare but lethal disease. These tumors tend to present at more advanced stages when compared to adult cases. Additionally, the inability to obtain accurate diagnosis often further delays the onset of treatment. Here, we present two cases of auricular melanoma in 3-year-old patients that were biopsied by the shave method prior to referral to an otolaryngologist. With the staging of the disease in question, sentinel lymph node biopsy were completed. Case 1 demonstrated positive sentinel nodes and underwent subsequent modified neck dissection. The second patient with negative sentinel nodes required no further surgery. Both patients remain disease free currently, at 16 and 22 months after the sentinel node biopsy. By discussing our experience and reviewing the current literature, we hope to shed some light in the diagnosis, staging and treatment of pediatric melanoma of the head and neck. PMID- 15364509 TI - Inflammatory bowel disease and sclerosing cholangitis with ulcerative lesions on skin and soft palate. AB - Described is a 15-year-old boy who presented with an ulcerative destructive soft palate lesion, skin lesions, and prolonged history of weakness and abdominal pain associated with abnormal liver functions. A midline destructive lesion was considered. However biopsies revealed chronic inflammation. After thorough investigation including colon, liver, skin and uvular biopsies a diagnosis of Inflammatory Bowel Disease in association of Autoimmune Sclerosing Cholangitis was made. Treatment with steroids improved his symptoms, normalized liver functions and enhanced healing of the skin and palatal lesions. This case demonstrates the involvement of the palate in a rare systemic disease and to our knowledge is the first description of Sclerosing Cholangitis presenting as an ulcerative lesion of the soft palate. PMID- 15364510 TI - Recruitment of benthic organisms onto a planned artificial reef: shifts in community structure one decade post-deployment. AB - Most artificial reef (AR) studies have examined the early colonization stages of benthic communities, while only a few have monitored the development of AR communities beyond the initial successional phases and evaluated the time scale needed for such development. In addition, despite the proliferation of AR studies, comparative studies between artificial and natural reefs (NRs) are scarce. We present here the monitoring results of initial (1-2 year) and progressed (10 year) stages of the developing benthic communities of a purpose planned AR submerged at Eilat, Israel (Red Sea), and compare them to its adjacent NR. Visual surveys of macro-invertebrates were conducted on the initial stages and coral communities were characterized at the progressed stage, using belt transects. The results demonstrate a distinct shift in species composition of the AR communities along the monitoring periods: from a soft coral dominated community, comprised mainly of Dendronephthya hemprichi, in initial developmental stages of up to two years post-deployment, to a community dominated by the sponge Crella cyatophora at year 10. Distinct differences in coral species count, living cover and diversity were found between the AR and its neighboring NR. We estimate the time frame required to develop a progressed diverse AR community to be well over a decade, even in tropical ecosystems. The factors shaping the species composition of purpose-designed ARs in a coral reef environment, including structural design, spatial orientation, depth and age, are discussed. PMID- 15364511 TI - Partitioning of polycyclic aromatic hydrocarbons between water and particles compared to bioaccumulation in mussels: a harbour case. AB - Water and particles from three sites located in Halifax Harbour, near and further away from combined municipal sewage effluents (CMSE) were analysed for parental and alkylated polycyclic aromatic hydrocarbons (par and alkPAH). Bioavailability of PAH was compared for inter-tidal mussels collected at the same sites in April and November. The PAH fingerprint determined over 9 sampling times covering a period of 19 months differed more between phases (water and particles) and seasons, than between sites. In the spring, more alkPAH associated with diesel and gasoline were detected in the soluble phase, along with more bioaccumulation of alkPAH in inter-tidal mussels. A broader number of parPAH were detected in mussels collected in the fall. The mean sum of dissolved alkPAH concentrations was higher in water at the site closer to raw CMSE than at the other two sites and particles of that site. However, lowest bioconcentration factors (BCF) were determined in mussels of this more contaminated site. Similar biota-particle accumulation factors (BPAF) were determined for parPAH in mussels from the three sites, all lower than the BCF of alkPAH. The study indicates that sewage treatment plants will reduce the amount of parPAH especially larger than fluoranthene and pyrene from being deposited in the harbour; that alk naphthalenes and fluorenes present in water will continue to be discharged; that the disturbance of sediments can make particle-bound PAH available to mussels. PMID- 15364512 TI - Metal concentrations in sediments and clams in four Moroccan estuaries. AB - Metals (Cd, Cu, Ni, Zn, Mn and Fe) were analyzed seasonally over three years in sediments and in tissues of the clam Scrobicularia plana in four Moroccan Atlantic estuaries: Loukkos, Sebou, Bou Regreg and Oum er Rbia. Of these metals, Cd was at the lowest concentrations in sediment. Concentrations of Cu, Zn, and to a lesser extent Ni, in sediments suggest greater contamination in Sebou and Bou Regreg than in the other estuaries. The fluctuations of Mn and Fe concentrations in the fine surface sediments reflect their continental origin and show seasonal variations that indicate soil run-off following rain events. Concentrations of Cu, Zn, and especially Ni in clam tissues in these estuaries were generally higher than in some other common bioindicator bivalve species. The seasonal variations in S. plana's tissue metal concentrations are linked to patterns of reproductive activity for all metals except Cd and possibly Zn, whose tissue concentrations may be regulated. Mn and Fe concentrations in S. plana were positively correlated to sediment levels of these metals. PMID- 15364513 TI - Comparison of polycyclic aromatic hydrocarbon distributions and sedimentary organic matter characteristics in contaminated, coastal sediments from Pensacola Bay, Florida. AB - In this study, we examined the distribution of polycyclic aromatic hydrocarbons (PAHs) in a contaminated coastal area and the characteristics of the natural organic matter in tandem. We present a detailed study of PAH concentration, distribution, and organic matter characteristics of three core samples from Pensacola Bay, Florida. Solid-state 13C Nuclear Magnetic Resonance (NMR), pyrolysis gas chromatography coupled with mass spectrometry (GC-MS), and tetramethyl ammonium hydroxide (TMAH) thermochemolysis GC-MS were applied to obtain structural details about the sedimentary organic matter. Elemental compositions (carbon and nitrogen) and estimates of black carbon contents are also reported. These coastal sediments were found to contain more PAHs in the upper 15 cm layers than in the bottom 15-25 cm samples. The samples that contained the most PAHs also contained the least amount of aromatic carbon and contained a significant amount of paraffinic carbon. Lignin-derived pyrolysis and TMAH thermochemolysis products were abundant and generally higher in all of the samples in comparison to those reported for modern coastal sediments, indicating a large flux of terrestrial carbon. The black carbon contents were found to range from 4.3% to 6.8%, which are significantly lower than other reports of black carbon in sediments, which represent as much as 65% of the total organic carbon content. The low black carbon content suggests that this type of refractory carbon may not be as responsible for regulating PAH distribution as indicated by other researchers. PMID- 15364514 TI - PCDD/F and PCB in harbour seals (Phoca vitulina) from British Columbia: response to exposure to pulp mill effluents. AB - Blubber of harbour seals (Phoca vitulina) sampled from the Strait of Georgia, BC, in 1991 and 1992 contained higher concentrations of polychlorinated dibenzo-p dioxins and furans (PCDD/F) than did samples from Quatsino Sound on western Vancouver Island. The higher proportions of 1,2,3,6,7,8-hexachlorodibenzodioxin and 2,3,7,8-tetrachlorodibenzofuran in the Strait of Georgia samples probably reflect contamination arising from the discharge of effluents from bleached kraft mills. Higher concentrations of non-ortho- and mono-ortho substituted polychlorinated biphenyls were also found in the Strait of Georgia samples, probably as a result of general industrial activity in the region. Principal components analysis showed that residues were distributed differently between males and females. Within males, concentrations of PCDD, PCDF and PCBs appeared to increase with age. Toxic equivalents (TEQ) to 2,3,7,8-tetrachlorodibenzodioxin in the Quatsino Sd. samples were close to the working guidelines for edible fish used by Health Canada; TEQ calculated for the Strait of Georgia samples exceeded these guidelines by up to a factor of five. PMID- 15364515 TI - Biofilm quantification on stone surfaces: comparison of various methods. AB - In the present study, the efficacy of three different methods of estimating biofilm biomass on stone (amount of chlorophyll a, fluorescein diacetate hydrolysis and changes in colour) is discussed. Stone samples were sprayed with solutions of cyanobacteria and the actual microbial biomass--determined by cell culture--was compared with the biomass estimated using the different methods. Statistically significant differences between actual biomass and that estimated from the fluorescein diacetate hydrolysis method invalidate its use. However, although the results indicated the amount of chlorophyll to be a good estimator of biofilm biomass, problems in the total extraction of chlorophyll from the stone cannot be overlooked. Determination of changes in colour was found to be a good method of quantifying biofilms growing on stone. PMID- 15364516 TI - Influence of Typha latifolia and fertilization on metal mobility in two different Pb-Zn mine tailings types. AB - Storing metal-rich mine waste (tailings) under submerged and reduced conditions can prevent the release of metals to the water column, but introduction of wetland plants on these sediments may alter the reducing environment through root oxygen diffusion or organic matter accumulation. Fertilization of these wetlands can enhance plant growth, but also may either strengthen reducing conditions via microbial stimulation, or increase the redox potential (Eh) through increased root radial oxygen loss. This long-term study (2.25 years) investigated the porewater As, Fe, and Zn concentrations of waterlogged Pb-Zn tailings from two Irish mines, Silvermines and Tara mines, with addition of Typha latifolia, fertilizer, or both treatments combined. In both tailings types, the fertilized plants showed significantly increased total biomass production, but the plants grew greater biomass in Tara tailings relative to Silvermines tailings even without fertilization. In Tara mines tailings, the addition of plants increased Eh and mobilized Zn; the addition of fertilizer enhanced reducing conditions and increased porewater concentrations of As and soluble sulfides; and the combination of treatments on these tailings resulted in complex interactions. In Silvermines tailings, there were negligible effects of the treatments. For effective sequestration of metals in these tailings, Silvermines would require only water cover, but Tara mines tailings would require either both treatments or neither because each treatment individually would increase solubility of As or Zn. These results show also the necessity of evaluating treatment effects specific to individual tailings, that long-term studies (years) are crucial for tailings equilibration and valid experimental conclusions, and that passive accumulation of organic matter may take decades. PMID- 15364517 TI - Evaluation of management strategies for reducing nitrogen loadings to four US estuaries. AB - In this study we used the Watershed Assessment Tool for Evaluating Reduction Strategies for Nitrogen (WATERSN) model to evaluate a variety of management strategies for reducing nitrogen (N) loads to four US east coast estuaries: Casco Bay, Long Island Sound, Chesapeake Bay and Pamlico Sound. These management strategies encompass reductions in atmospheric emissions and deposition of N from sources including, fossil fuel burning utility emissions and mobile NO(x) emissions, N treatment in wastewater and controls on agricultural N inputs. We find that in primarily urban watersheds biological removal of N in wastewater treatment produces the greatest reduction in N loading (32-57% reductions), while in less urban watersheds, reductions in agricultural loading are more effective (5-56% reductions) in decreasing N loads to coastal ecosystems. Because anthropogenic N inputs are derived from a variety of sources, we also examined an integrated scenario targeting all major N sources; this resulted in 35-58% reductions in N loading. Nitrogen pollution originates from multiple sources and is transported through several media (air, soil, water); a major challenge of the development of N management strategies will be the control of multiple sources to effectively reduce N loads to estuaries. PMID- 15364518 TI - Treatment of pulp and paper mill wastewater--a review. AB - Pulp and paper mills generate varieties of pollutants depending upon the type of the pulping process. This paper is the state of the art review of treatability of the pulp and paper mill wastewater and performance of available treatment processes. A comparison of all treatment processes is presented. Combinations of anaerobic and aerobic treatment processes are found to be efficient in the removal of soluble biodegradable organic pollutants. Color can be removed effectively by fungal treatment, coagulation, chemical oxidation, and ozonation. Chlorinated phenolic compounds and adsorable organic halides (AOX) can be efficiently reduced by adsorption, ozonation and membrane filtration techniques. PMID- 15364519 TI - Variations in the source, metal content and bioreactivity of technogenic aerosols: a case study from Port Talbot, Wales, UK. AB - Atmospheric aerosol samples were collected during different prevailing wind directions from a site located close to a busy motorway, a major steelworks, and the town of Port Talbot (Wales, UK). A high-volume collector was used (1100 l/min), enabling relatively large amounts of particulate matter (PM(10-2.5) and PM(2.5)) samples to be obtained on a polyurethane foam [PUF, H(2)N-C(O)O CH(2)CH(3)] substrate over periods of 2-7 days. Four samples were chosen to exemplify different particle mixtures: SE- and NE-derived samples for particles moving along and across the motorway, a NW-derived sample from the town, and a mixed SW/SE-derived sample containing a mixture of particles from both steelworks and motorway. The latter sample showed the highest average collection rate (0.9 mg/h, 13 microg/m(3)) and included a prominent pollution episode when rainy winds were blowing from the direction of the steelworks. Both NW and SE samples were collected under dry conditions and show the same collection rate (0.7 mg/h, 10 microg/m(3)), whereas the NE sample was collected during wetter weather and shows the lowest rate (0.3 mg/h, 5 microg/m(3)). Scanning electron microscopy (SEM) and energy-dispersive X-ray microanalysis system (EDX) analyses show all samples are dominated by elemental and organic carbon compounds (EOCC) and nitrates, with lesser amounts of sulphates, felsic silicates, chlorides and metals. ICP-MS analyses show the SW/SE sample to be richest in metals, especially Fe, Zn, Ni, and Mn, these being attributed to an origin from the steelworks. The SE sample, blown along the motorway corridor, shows enhanced levels of Pb, V, Ti, As, and Ce, these metals being interpreted as defining a traffic-related chemical fingerprint. The NW sample shows a very low metal content. DNA plasmid assay data on the samples show TM(50) values varying from 66 to 175 microg/ml for the adjusted whole sample and 89 to 203 microg/ml for the soluble fraction. The SW/SE mixed metalliferous sample is the most bioreactive (both whole and soluble) and the soluble fraction of the metal-depleted NW sample is the least bioreactive. The metal content of the aerosol samples, especially soluble metals such as Zn, is suggested to be the primary component responsible for oxidative damage of the DNA, and therefore most implicated in any health effects arising from the inhalation of these particulate cocktails. PMID- 15364520 TI - The potential impact of soil ingestion on human mineral nutrition. AB - Geophagia, the intentional and repeated ingestion of soil material, is a complex eating behaviour with incomprehensible aetiology. It is generally assumed that geophagia may help supplement mineral nutrients and thus should not be dissuaded, particularly in subsistence communities. This is largely based on the assumption that a large proportion of mineral nutrients in geophagic materials is potentially available for absorption in the body. We tested this hypothesis on five soils collected from geophagia-prevalent communities by using an in vitro soil ingestion simulation test that is broadly similar to the gastrointestinal (GI) tract. The results show that, despite the soils being rich in mineral nutrients, soil ingestion, inadvertent or through geophagia can potentially reduce the absorption of already bioavailable nutrients, particularly micronutrients such as Fe, Cu and Zn. These in vitro findings, while disagreeing with the commonly held view of geophagia as a source of nutrient supplementation, are consistent with micronutrient deficiency problems observed in clinical nutrition studies conducted amongst geophagic populations. The work also showed that, in some cases, the ingested soils may become a source of Ca, Mg and Mn, although it is not clear why other similar soils should not release any of these mineral nutrients. PMID- 15364521 TI - Fish availability in supermarkets and fish markets in New Jersey. AB - There is considerable interest in fish consumption, contaminant loads in edible fish, and the risk from consuming fish. Both the benefits and the risks from eating fish are publicized. Most of this attention has focused on recreational anglers and self-caught fish, although the vast majority of fish that people eat are purchased from commercial sources: fish markets and supermarkets. We examined the availability of fish in supermarkets and specialty fish markets in New Jersey, including three regions of the state in communities with high and low per capita incomes (upscale vs. downscale neighborhoods). We were particularly interested in examining whether consumers could determine what type of fish they were buying and whether it was farm-raised or wild. Flounder and salmon were the most commonly available fish, followed by bluefish and tilapia. There were few significant differences in the availability of fish as a function of region. Fish were equally available in fish markets and supermarkets, although snappers were more available in fish markets. The most common fish (found in over 60% of stores) were equally available in upscale and downscale neighborhood stores. However, there were some significant differences in less common fish; butterfish, croaker, monkfish, porgy, and whiting were more available in downscale markets, and halibut, sole, and swordfish were more available in upscale markets. Information available to consumers on labels varied markedly: (1) most labels were generic but some indicated species (e.g., Spanish vs. Boston mackerel, Chilean vs. Black sea bass, mako vs. black-tip shark, rainbow vs. steelhead trout); (2) in many cases, labels indicated whether catfish or salmon were farmed or wild, but usually that information was lacking; (3) sometimes, the labels indicated the location where fish were caught (salmon); and (4) sometimes, there was information on both species and type (e.g., farm/wild for salmon). In most cases, labels gave only a fish name and price. Consumers would be able to make more informed choices if the provenance of fish was clearly stated. State agencies might improve information available to consumers by providing distributors and markets with guidelines about the types of information necessary for consumers to make informed decisions about the fish they eat. When asked, counter staff often could not answer where fish originated from. Finally, there should be partnerships between government agencies responsible for public health, risk assessors, and consumers to ascertain the types of information consumers want and to provide the best available information to consumers. PMID- 15364522 TI - Is the digestive gland of Mytilus galloprovincialis a tissue of choice for estimating cadmium exposure by means of metallothioneins? AB - A study performed over 12 months with caged mussels Mytilus galloprovincialis in the coastal marine zone, which is under urban pressure, reveals a temporal variation of digestive gland mass, which causes "biological dilution" of cytosolic metallothionein (MT) and trace metal (Cd, Cu, Zn, Fe, Mn) concentrations. The dilution effect was corrected by expressing the cytosolic MT and metal concentrations as the tissue content. Consequently, the changes of the average digestive gland mass coincide with the changes of MT and trace metal contents. From February to June, MT contents are nearly twice and trace metal contents nearly three times higher than those of the other months. The period of increased average digestive gland mass, of MT and trace metal contents probably overlaps with the sexual maturation of mussels (gametogenesis) and enhanced food availability. Since natural factors contribute more to the MT content than the sublethal levels of Cd, the digestive gland of M. galloprovincialis is not considered as a tissue of choice for estimating Cd exposure by means of MTs. PMID- 15364523 TI - Heavy metal bioaccumulation and macroinvertebrate community changes in a Mediterranean stream affected by acid mine drainage and an accidental spill (Guadiamar River, SW Spain). AB - In the 1998 Aznalcollar mining accident, 5 million cubic meters of toxic waste were spilled into the Guadiamar River (SW Spain). This paper describes the likely effects of metal pollution on the macroinvertebrate community in the first 15 km of the river adjacent to the mine, 2 years after the spill. The contents of Zn, Cu, Pb, As, Cd, Tl, and Sb in water, sediment, and in the caddisfly Hydropsyche were analysed, together with some physico-chemical parameters. From the results of physico-chemical and community parameters, the studied area was divided into three sections: the first section contained upstream control stations, not affected by current mining activities, and with a diverse macroinvertebrate community structure (mean number of families = 19); the second section, close to the mine, was severely polluted, with more than 50 mg l(-1) of Zn or 1.6 mg l(-1) of Cu in water, a pH of nearly 4, and a very diminished macroinvertebrate community (three families); in the last section, 6-15 km downstream from the mine, the water quality improved, metal concentrations decreased, pH reached neutrality and the macroinvertebrate community comprised eight families. We also determined metal concentrations in the larvae of Hydropsyche and found increases of 3- to 35-fold (for Cu and Cd, respectively) compared to control sites. As far as we are aware, these values are the highest concentrations so far detected in this caddisfly anywhere in the world. Different patterns of metal and pH tolerance were identified in macroinvertebrates. Most of the species were intolerant to present pollution levels and were only present in control sites. Others were tolerant to metal pollution but not to low pH and were present in the main river downstream of the mine, while a very few species managed to live in the most polluted section, with very low pH and high metal concentrations. PMID- 15364524 TI - Comet and micronucleus assays in zebra mussel cells for genotoxicity assessment of surface drinking water treated with three different disinfectants. AB - The aim of this research was to study the influence of classic (sodium hypochlorite and chlorine dioxide) and alternative (peracetic acid [PAA]) disinfectants on the formation of mutagens in surface waters used for human consumption. For this proposal, in vivo genotoxicity tests (Comet and micronucleus assay) were performed in an experimental pilot plant set up near Lake Trasimeno (Central Italy). The effects were detected in different tissues (haemocytes for the Comet assay and gills for the micronucleus test [MN]) of Dreissena polymorpha exposed in experimental basins supplied with lake water with/without the different disinfectants. Specimen collection was performed before disinfectant input for both tests and after the start of disinfection (3 h and 20 days for the Comet assay and 10 and 20 days for micronucleus test, respectively) to assess short- and long- term exposure effects during three sampling campaigns (October 2000, February 2001, and June 2001). Seasonal differences in baseline levels of DNA migration and micronucleus frequency were observed. Raw water quality modulation on disinfection by-product formation was shown. The results of the micronucleus and Comet assays on zebra mussel cells after in situ exposure to water disinfected with the two chlorinated compounds clearly indicate DNA/by-product interaction. PAA did not induce either clastogenic/aneugenic effects or DNA damage on this bioindicator. PMID- 15364525 TI - Pharmaceuticals and personal care products (PPCPs) and endocrine disrupting chemicals (EDCs) in stormwater canals and Bayou St. John in New Orleans, Louisiana, USA. AB - Samples were collected from two stormwater canals and a recreational urban waterway known as Bayou St. John in New Orleans, Louisiana, USA and analyzed for a range of pharmaceuticals and personal care products (PPCPs) and endocrine disrupting chemicals (EDCs). Concentrations of 7 PPCPs and EDCs were measured by a method that provides for simultaneous extraction and quantification of the following compounds: clofibric acid, naproxen, ibuprofen, fluoxetine, clorophene, triclosan, bisphenol A. The method also was used as an indicator of the occurrence of estrogenic compounds by targeting estrone and 17beta-estradiol. The two canals (Orleans and London) are used to drain a portion of the city's stormwater directly into the Mississippi River or Lake Pontchartrain. Bayou St. John is located between the two canals and supplied with water from Lake Pontchartrain. Results from the 6-month sampling period indicated the following concentration ranges for the two stormwater canals: naproxen (ND - 145 ng/l), ibuprofen (ND - 674 ng/l), triclosan (ND - 29 ng/l) and bisphenol A (1.9-158 ng/l). Concentrations of these target analytes increased with cumulative rainfall. For bayou waters, only naproxen (2.1-4.8 ng/l) and bisphenol A (0.9-44 ng/l) were detected. Estrone was detected but determined non-quantifiable for multiple sampling events at the 3 sites. None of the other target analytes (clofibric acid, fluoxetine, clorophene, and 17beta-estradiol) were detected above their method detection levels. Results of this study demonstrate the occurrence of PPCPs and EDCs in New Orleans stormwater canals and Bayou St. John. Results also demonstrate the use of this analytical technique as an indicator of non-point source sewage contamination in New Orleans stormwater canals. PMID- 15364526 TI - Contaminant levels in sediments and asteroids (Asterias rubens L., Echinodermata) from the Belgian coast and Scheldt estuary: polychlorinated biphenyls and heavy metals. AB - The Southern Bight of the North Sea is particularly exposed to anthropogenic contamination, due to heavy urbanisation and industrialisation of its catchment area. The present work focuses on polychlorinated biphenyl (PCB) and metal contamination of the marine environment along and off the Belgian coast. Its objectives were to compare the concentrations of seven PCB congeners and four heavy metals in the sediments (a repository for anthropogenic contaminants) and in the asteroid Asterias rubens (a recognized bioindicator species). Nineteen sampling stations were considered between the mouth of the Scheldt Estuary and the southern limit of the Belgian coast (asteroids were found in 10 out of the 19 stations). PCB and metal concentrations measured in sediments and asteroids were in the range of values reported in previous studies. Stations under direct influence of the Scheldt were the most impacted by the considered contaminants. Metal concentrations varied according to the grain-size fraction considered. In asteroids, PCBs and metals were found to be selectively distributed among body compartments, and pyloric caeca were found to most efficiently discriminate between sampling stations contamination levels. PCB and metal analysis of sediments provided a physicochemical evaluation of the contamination, whereas analysis of asteroids introduced a biological dimension to the approach by taking into account bioavailability of the contaminants. PMID- 15364527 TI - Investigating the environmental transport of human pharmaceuticals to streams in the United Kingdom. AB - The occurrence of 12 selected pharmaceutical compounds and pharmaceutical compound metabolites in sewage treatment works (STW) effluents and surface waters was investigated. The substances selected for the monitoring programme were identified by a risk ranking procedure to identify those substances with the greatest potential to pose a risk to the aquatic environment. STW final effluent and surface water samples were collected from Corby, Great Billing, East Hyde, Harpenden and Ryemeads STWs. Ten of the 12 pharmaceutical compounds were detected in the STW effluent samples: propranolol (100%, median = 76 ng/l), diclofenac (86%, median = 424 ng/l), ibuprofen (84%, median = 3086 ng/l), mefenamic acid (81%, median = 133 ng/l), dextropropoxyphene (74%, median = 195 ng/l), trimethoprim (65%, 70 ng/l), erythromycin (44%, < 10 ng/l), acetyl sulfamethoxazole (33%, median =< 50 ng/l), sulfamethoxazole (9%, median =< 50 ng/l), tamoxifen (4%, median =< 10 ng/l). In the corresponding receiving streams, fewer compounds and lower concentrations were found: propranolol (87%, median = 29 ng/l), ibuprofen (69%, median = 826 ng/l), mefenamic acid (60%, median = 62 ng/l), dextropropoxyphene (53%, median = 58 ng/l), diclofenac (47%, median =< 20 ng/l), erythromycin (38%, median =< 10 ng/l), trimethoprim (38%, median =< 10 ng/l), acetyl sulfamethoxazole (38%, median =< 50 ng/l). Four human pharmaceutical compounds were detected in samples upstream of the STWs sampled: ibuprofen (57%, median = 181 ng/l), trimethoprim (36%, median < 10 ng/l), erythromycin (17%, median =< 10 ng/l), propranolol (14%, median =< 10 ng/l), suggesting that longer range stream transport of some compounds is possible. The particular STW that was sampled and the month that it was sampled significantly influenced the measured concentrations of several, but not all, substances. There was no significant relationship between usage data and the overall frequency with which different substances were detected. There was however, some evidence to suggest that usage data are positively associated with concentrations of pharmaceuticals in effluent and, particularly, with concentrations measured in surface waters below STWs. These results suggest that most sewage treatment works in England and Wales are likely to be routinely discharging small quantities of pharmaceuticals into UK rivers. None of the pharmaceuticals were found at concentrations that were high enough to cause acute toxic impacts to aquatic organisms. However, insufficient data were available to be able to comment on whether the concentrations measured have the potential to result in more subtle long-term effects on aquatic organisms (e.g. effects on growth, ability to reproduce). PMID- 15364528 TI - Monitoring of selected estrogenic hormones and industrial chemicals in groundwaters and surface waters in Austria. AB - In order to address the Austrian situation concerning endocrine disrupting substances (EDS), a consortium called Austrian Research Cooperation on Endocrine Modulators (ARCEM) was established in 1999. Among several other issues that were investigated, ARCEM monitored more than 400 ground- and surface water samples for selected estrogenic hormones and industrial chemicals. Appropriate analytical methods were established using GC-HRMS for the detection of hormones and LC/MSMS for the detection of industrial chemicals. Since analytical results were forwarded for toxicological assessments within the programme, quantification limits below 0.1 ng/l (ethinyl estradiol) and 10 ng/l (industrial chemicals) were required depending on the NOEL of the individual compound. Results indicate that both hormones and industrial chemicals occur in the selected ground- and surface water sites in detectable concentrations. Three river sites were identified as being charged with noticeable concentrations of EDS. These results were verified by biological indications within the part of the ARCEM programme dealing with bio indications. PMID- 15364529 TI - Evaluation of methane emissions from Taiwanese paddies. AB - The main greenhouse gases are carbon dioxide, methane and nitrous oxide. Methane is the most important because the warming effect of methane is 21 times greater than that of carbon dioxide. Methane emitted from rice paddy fields is a major source of atmospheric methane. In this work, a methane emission model (MEM), which integrates climate change, plant growth and degradation of soil organic matter, was applied to estimate the emission of methane from rice paddy fields in Taiwan. The estimated results indicate that much methane is emitted during the effective tillering and booting stages in the first crop season and during the transplanting stage in the second crop season in a year. Sensitivity analysis reveals that the temperature is the most important parameter that governs the methane emission rate. The order of the strengths of the effects of the other parameters is soil pH, soil water depth (SWD) and soil organic matter content. The masses of methane emitted from rice paddy fields of Taiwan in the first and second crop seasons are 28,507 and 350,231 tons, respectively. The amount of methane emitted during the second crop season is 12.5 times higher than that emitted in the first crop season. With a 12% reduction in planted area during the second crop season, methane emission could be reduced by 21%. In addition, removal of rice straw left from the first crop season and increasing the depth of flooding to 25 cm are also strategies that could help reduce annual emission by up to 18%. PMID- 15364530 TI - Evaluation of the genotoxic, mutagenic and oxidant stress potentials of municipal solid waste incinerator bottom ash leachates. AB - Triplicate aqueous leachates of a municipal solid waste incineration bottom ash (MSWIBA) were produced according to a European standardised method. Leachates analysis showed relatively low concentrations (less than 1 mg.l(-1)) for four metals (iron, cadmium, lead and copper). No mutagenic activity was revealed after performing the Salmonella/microsome assay with and without microsomal activation. With the Vicia root tip micronucleus assay, a significant increase in micronucleated cells was observed between 3.4% and 100% leachate concentration. Significant and elevated antioxidant stress enzyme activities, e.g., superoxide dismutase (SOD), catalase (CAT), peroxidase (PER) and glutathione reductase (GR), were detected in Vicia root tissues even at the lowest tested leachate concentration (i.e., 0.3%), whereas this was not always the case in leaf tissues, which showed tissue specificity for the tested enzymes. At the lowest concentration (i.e., 0.3%), a higher increase was observed (respectively 197% and 45% compared to the control) for root glutathione reductase and peroxidase activities over those of other enzymes (superoxide dismutase and catalase). Our results suggest that MSWIBA aqueous leachates need to be formally tested with genotoxic sensitive tests before recycling and support the hypothesis that plant genotoxicity is related to the cellular production of reactive oxygen species (ROS). PMID- 15364531 TI - Characterization of dissolved organic matter from source to sea using fluorescence and absorbance spectroscopy. AB - Dissolved organic matter fluorescence, absorbance and dissolved organic carbon were measured from source to sea in the River Tyne catchment, of approximately 2935 km(2) and encompassing areas of contrasting land use. The catchment has three major tributaries: the North Tyne which has good water quality, high dissolved organic carbon concentrations and visible water colour from the high proportion of peat in its upper catchment; the South Tyne which has good water quality with typical riverine dissolved organic carbon concentrations and drains from limestone uplands; and the Derwent, a more urbanized catchment which is increasingly impacted by treated sewage effluent discharges towards its mouth. Thirty sample sites, 23 along the three main tributaries and seven within the estuary, were sampled on six occasions over the period 2002-2003. High absorbance at 340 nm and dissolved organic carbon concentration identify N Tyne waters due to the peaty headwaters, but no downstream trends in these parameters are observed in any of the tributaries, in contrast to the estuary where a rapid decrease is observed in both. Fluorescence in contrast demonstrated downstream trends in both intensity and wavelength, especially in the Derwent as it is increasingly impacted by anthropogenic dissolved organic matter. Elevated protein like fluorescence intensity also fingerprints sewage effluent within the estuary. The absorbance coefficient at 340 nm was found to have the strongest correlation to dissolved organic carbon concentration, greater than all fluorescence intensity parameters measured. However, fluorescence analysis permits the source of the dissolved organic matter to be determined, and therefore has implications for understanding its fate in estuaries and the ocean. PMID- 15364532 TI - Assessment of acid neutralizing capacity and potential mobilisation of trace metals from land-disposed dredged sediments. AB - In the past decades, large amounts of contaminated sediments have been dredged and spread out along rivers. The understanding of the fate and the behaviour of contaminants in dredged sediments is essential to deal with the management of contaminated sediments. Heavy metal leaching behaviour in dredged sediments of different ages since disposal, varying from 10 to 70 years, was assessed by pH(stat) leaching test and potentiometric titrations. Based on the pH(stat) titration results, two operationally defined pools of heavy metals, i.e., a labile and a slowly labile pool of heavy metals, were distinguished and quantified by mathematical description of leaching behaviour. The mathematical description of leaching behaviour during pH(stat) and potentiometric titrations allowed deduction of four groups of elements with a different type of leaching behaviour. Considering heavy metals, Zn, Cd and Ni displayed a leaching behaviour similar to acid neutralizing capacity (ANC), whereas the release of Cu, Pb and Cr was slower than ANC. However, no significant differences with regard to heavy metal leaching behaviour were found between sediments with different time since disposal on land. The elevated acid neutralizing capacity of the dredged sediments analysed in this study strongly decrease the risk of soil acidification with associated heavy metal leaching. The prediction of soil acidification over an extended period of time, together with the potential release of trace metals, was considered an important advantage of pH(stat) leaching tests compared to conventional leaching procedures. PMID- 15364533 TI - Potential biogeochemical and ecological development of a flooded tailings impoundment at the Kristineberg Zn-Cu mine, northern Sweden. AB - The potential short-term (10(2) years) biogeochemical and ecological effects of diverting stream water (pH 4.9-6.7) into a limed, flooded tailings impoundment (pH 8-12) were studied by combining geochemical and biological data. In the long-term perspective, the successional development of lakes was used as a natural analogue. Based on the vertical distribution of temperature and total dissolved solids (TDS < 0.22 microm), the impoundment can be characterised as a continuous/discontinuous cold polymictic lake, with holomictic summer circulation. A re-inoculation study indicated that the growth of autotrophic, aerobic bacteria (presumably Acidithiobacillus ferrooxidans) presently is inhibited by the high pH in the impoundment. In a short-term perspective, termination of liming and diversion of stream water into the impoundment will result in a complex interplay between physical, biogeochemical and ecological effects. A reduced vertical mixing of the approximately 2-m-deep water column, dissolution of calcite and gypsum (compounds of a sludge formed in the impoundment) and an enhanced microbiological activity are major expected effects. The dissolution of calcite may act as a pH buffer and result in metal remobilisation from the sludge. Excluding autochthonous organic matter produced in the impoundment, streamwater input of suspended matter and formation of settling flocculants are expected to result in a sediment accumulation rate of approximately 1.5 mg cm(-2) year(-1) (1.6-3.3 cm/10(2) years). Settling allochthonous organic C (0.15-0.30 mg C cm(-2) year(-1)) may serve as an oxygen barrier and as a reservoir of organic compounds capable of driving redox reactions. In a long-term perspective, a hydroseral development into a wetland/peatland can be expected, with a bog lake, poor fen or flat bog as final stage. This development presupposes a decreasing pH when liming is terminated and stream water is diverted into the impoundment. It also assumes that the impoundment will be similar to an acidified lake, and that the succession is driven by Sphagnum colonizing the impoundment. If the hydrological conditions/water level is affected (e.g., by climatic changes or a dam failure), a terrestrialization culminating in coniferous forest on peat soil may occur. PMID- 15364534 TI - Seasonal water quality variations in a river affected by acid mine drainage: the Odiel River (South West Spain). AB - This paper intends to analyse seasonal variations of the quality of the water of the Odiel River. This river, together with the Tinto River, drains the Iberian Pyrite Belt (IPB), a region containing an abundance of massive sulphide deposits. Because of mining activity dating back to prehistoric times, these two rivers are heavily contaminated. The Odiel and Tinto Rivers drain into a shared estuary known as the Ria of Huelva. This work studies dissolved contaminant data in water of the Odiel River collected by various organisations, between October 1980 and October 2002, close to the rivers entry into the estuary. Flow data for this location were also obtained. The most abundant metals in the water, in order of abundance, are zinc (Zn), iron (Fe), manganese (Mn) and copper (Cu). Arsenic (As), cadmium (Cd) and lead (Pb) are also present but in much lower quantities. The quality of the river water is linked to precipitation; the maximum sulphate, Fe, Zn, Mn, Cd and Pb concentrations occur during the autumn rains, which dissolve the Fe hydroxysulphates that were precipitated during the summer months. In winter, the intense rains cause an increase in the river flow, producing a dilution of the contaminants and a slight increase in the pH. During spring and summer, the sulphate and metal concentration (except Fe) recover and once again increase. The Fe concentration pattern displays a low value during summer due to increased precipitation of ferric oxyhydroxides. The arsenic concentration displays a different evolution, with maximum values in winter, and minimum in spring and summer as they are strongly adsorbed and/or coprecipitated by the ferric oxyhydroxides. Mn and sulphates are the most conservative species in the water. Relative to sulphate, Mn, Zn and Cd, copper displays greater values in winter and lower ones in summer, probably due to its coprecipitation with hydroxysulphates during the spring and summer months. Cd and Zn also appear to be affected by the same process, although to a lower degree than Cu, experiencing a slight reduction in summer with respect to Mn and sulphates. PMID- 15364535 TI - Effect of season of application on the adhesion, retention and recontamination of herbage by potentially toxic metals and sewage sludge. AB - Grassland in the United Kingdom can accept sewage sludge when, as a result of cropping restrictions, arable land cannot. However, one of the uncertainties associated with the application of sewage sludge to grassland is the potential exposure of the grazing ruminant to sewage sludge particulates adhered to the grass or deposited on the soil surface. The implications of elevated levels of sludge particulates and associated potentially toxic metals (PTMs) are the potential accumulation of PTMs into edible tissues of the grazing animal. Two experiments were performed to investigate the effect of season of application on the adhesion, retention and recontamination of herbage by sewage sludge and PTMs. In each experiment, three treatments were installed, an untreated control, application of sludge in the autumn and application of sludge in the spring. Sewage sludge particulates and PTMs were eluted sequentially 21 days after the application of sludge using water and detergent and the residual (tissue) concentrations were monitored. Total concentrations in each fraction and adhesion indices were calculated. Soil samples taken from the sward treatments after each application showed no elevated concentrations of PTMs or concentrations that approached the maximum concentrations allowed for soils amended with sewage sludge. The amounts of sludge adhered to herbage ranged from 114 to 360 g sludge DM/kg herbage DM. There was evidence of preferential retention of PTMs to herbage. However, during the 21-day no grazing period there was a general decline in concentration of all PTMs. Copper was retained on the herbage for greater periods of time compared to Zn, CD, Pb and Fe (P < 0.01). The proportion of Pb recovered from the water phase increased during the 21-day no grazing period in comparison to Cu, Zn and Fe (P < 0.001). It is concluded that the concentrations of PTMs adhered and integral to herbage as a result of surface application of sewage sludge are likely to fall to acceptable levels in well-managed permanent grassland. PMID- 15364536 TI - Liver and kidney concentrations of vanadium in oiled seabirds after the Erika wreck. AB - Vanadium was measured by atomic absorption spectrophotometry in the liver and kidney of several oiled seabirds that were stranded on the French Atlantic coasts after the Erika wreck and died in wildlife care Centers. Estimated averages were 30 to 77 ng g(-1) wet weight (ww) in the liver and 52 to 72 ng g(-1) wet weight in the kidney. These concentrations were not higher in oiled birds than in dead birds found later, without visible traces of petroleum on beaches not affected by the Erika pollution. Vanadium hepatic and renal concentrations do not seem to be appropriate biomarkers for recent exposure to fuel in seabirds. PMID- 15364537 TI - The effects of gender, age, ethnicity, and liver cirrhosis on cytochrome P450 enzyme activity in human liver microsomes and inducibility in cultured human hepatocytes. AB - We have measured cytochrome P450 (CYP) activity in nearly 150 samples of human liver microsomes and 64 samples of cryopreserved human hepatocytes, and we have performed induction studies in over 90 preparations of cultured human hepatocytes. We have analyzed these data to examine whether the expression of CYP enzyme activity in liver microsomes and isolated hepatocytes or the inducibility of CYP enzymes in cultured hepatocytes is influenced by the gender, age, or ethnicity of the donor (the latter being limited to Caucasians, African Americans, and Hispanics due to a paucity of livers from Asian donors). In human liver microsomes, there were no statistically significant differences (P > 0.05) in CYP activity as a function of age, gender, or ethnicity with one exception. 7 Ethoxyresorufin O-dealkylase (CYP1A2) activity was greater in males than females, which is consistent with clinical observation. Liver microsomal testosterone 6beta-hydroxylase (CYP3A4) activity was slightly greater in females than males, but the difference was not significant. However, in cryopreserved human hepatocytes, the gender difference in CYP3A4 activity (females = twice males) did reach statistical significance, which supports the clinical observation that females metabolize certain CYP3A4 substrates faster than do males. Compared with those from Caucasians and African Americans, liver microsomes from Hispanics had about twice the average activity of CYP2A6, CYP2B6, and CYP2C8 and half the activity of CYP1A2, although this apparent ethnic difference may be a consequence of the relatively low number of Hispanic donors. Primary cultures of hepatocytes were treated with beta-naphthoflavone, an inducer of CYP1A2, phenobarbital or rifampin, both of which induce CYP2B6, CYP2C9, CYP2C19, and CYP3A4, albeit it to different extents. Induction of these CYP enzymes in freshly cultured hepatocytes did not appear to be influenced by the gender or age of the donor. Furthermore, CYP3A4 induction in hepatocytes isolated from cirrhotic liver was comparable to that in normal hepatocytes, which supports the "healthy hepatocyte, sick environment" hypothesis of liver cirrhosis. This review summarizes these findings and discusses their implications for the use of human liver microsomes and hepatocytes for in vitro studies of drug metabolism and enzyme induction, which play a key role in drug development. PMID- 15364538 TI - Expression of cytochromes P450 1A1 and 1B1 in human lung from smokers, non smokers, and ex-smokers. AB - Cytochromes P450 1A1 and 1B1 are known to bioactivate procarcinogens such as polycyclic aromatic hydrocarbons (PAHs) found in cigarette smoke and are inducible via an Ah receptor-mediated mechanism. The aim of this study was to examine the levels of expression of CYP1A1 and CYP1B1 in samples of lung from smokers (n = 18), non-smokers (n = 7), and ex-smokers (n = 7). Using immunoglobulin preparations of highly specific polyclonal antibodies and immunoblot analysis of microsomes from lung tissues, we determined the specific content for CYP1A1 and CYP1B1. For CYP1A1, we found median expression levels of 15.5 pmol/mg microsomal protein in smokers, 6.0 pmol/mg microsomal protein in non smokers, and 19.0 pmol/mg microsomal protein in ex-smokers. The difference in median expression levels of smokers and ex-smokers compared to non-smokers was statistically significant. For CYP1B1, we found median expression levels of 1.8 pmol/mg microsomal protein in smokers, 1.0 pmol/mg microsomal protein in non smokers, and 4.4 pmol/mg microsomal protein in ex-smokers. The difference in median expression levels between ex-smokers and non-smokers was statistically significant. These results suggest that levels of expression of CYP1A1 and CYP1B1 protein in lung tissues from smokers and ex-smokers are quantitatively greater than in non-smokers. By immunohistochemical analysis, we demonstrated the expression of CYP1A1 and CYP1B1 in normal human alveolar type I and II cells, ciliated columnar epithelial cells lining bronchoalveolar airways, and alveolar macrophages. These results confirm that CYP1A1 is expressed in normal human lung, appears to be induced in smokers, and show interindividual variation; the similar characteristics of CYP1B1 are demonstrated. PMID- 15364539 TI - Disruption of cell-cell contact maximally but transiently activates AhR-mediated transcription in 10T1/2 fibroblasts. AB - The aryl hydrocarbon receptor (AhR) is activated by 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD), but activation without an exogenous ligand also occurs when normal cell-cell contact is prevented. Suspension of several C3H10T1/2 fibroblast clonal sub-lines that contain an integrated AhR-responsive reporter produced a time course and level of reporter activation and CYP1B1 induction that paralleled TCDD stimulation in confluent monolayer culture. Suspension activation was, however, more transient. Loss of cell-cell contact at low density also activated these reporters independent of cell cycle changes to levels comparable to TCDD stimulation of confluent cells. Loss of cell-cell contact may, therefore, activate AhR. Suspension and TCDD activations exhibited comparable nuclear translocation of AhR and then AhR/ARNT complex formation. Each AhR activation process was equally attenuated by inhibition of, respectively, HSP90 ATPase, the 26S proteosome, and by depletion of intracellular Ca2+. By contrast, the AhR antagonist alpha-naphthoflavone (alphaNF) blocked ligand-stimulated AhR activity, but not activation through loss of cell-cell contact. Suspension-induced reporter activation was selectively enhanced by LiCl, which prevented GSK-3beta effects on the simultaneously released beta-catenin. The effects of suspension and LiCl on reporters were reversed by Ro-31-8220, which did not affect beta-catenin, TCDD activation processes, or AhR turnover. Neither LiCl nor Ro-31-8220 altered suspension-induced AhR/ARNT complex formation. Loss of cell-cell contact permits nuclear translocation and AhR activation that is largely replicated after TCDD binding, but with activity differences due to contact-sensitive factors functioning after AhR/ARNT complex formation. PMID- 15364540 TI - Two distinct classes of CCAAT box elements that bind nuclear factor-Y/alpha actinin-4: potential role in human CYP1A1 regulation. AB - A negative regulatory element (NRE1; position -794 to -774) was previously identified that mediates the downregulation of CYP1A1, including partial suppression of Ah receptor-dependent induction. The CCAAT-box binding protein, nuclear factor-Y (NF-Y), is a component of one of two protein complexes that specifically and competitively bind the CYP1A1 NRE1 in vitro with nearly equal affinity. The second complex involves an unidentified protein(s) called the negative regulatory factor (NRF). Competitive electrophoretic mobility shift assays (EMSA) revealed two distinct classes of NF-Y-binding CCAAT-box elements distinguished by their ability or inability to also bind NRF. To further explore the identity of NRE1-binding proteins, a purification scheme was developed culminating in NRE1-dependent DNA affinity chromatography and sequence analysis. An approximate 106-kDa protein was purified and shown to be alpha-actinin-4 (ACTN4), one of two ubiquitously expressed non-muscle actinins. Electrophoretic mobility shift assays combined with Western blot analysis and co immunoprecipitation experiments suggested that ACTN4 is associated with the NF-Y complex, but not NRF. Attempts to demonstrate a role for NF-Y/ACTN4 in regulating CYP1A1 expression were unsuccessful, likely due to an inability to significantly change nuclear ACTN4 levels with phosphatidylinositol 3'-kinase agonists and antagonists. However, given ACTN4's known functions and the suspected functions of actin and actin-related proteins in chromatin remodeling and other nuclear events, ACTN4 may assist NF-Y in recruiting chromatin-remodeling complexes or may direct NF-Y/ACTN4-targeted genes to the nuclear matrix and active transcriptional complexes. PMID- 15364541 TI - PXR (NR1I2): splice variants in human tissues, including brain, and identification of neurosteroids and nicotine as PXR activators. AB - To gain insight on the expression of pregnane X receptor (PXR), we analyzed PXR.1 and PXR alternatively spliced transcripts in a panel of 36 human tissues. PXR.1 was expressed in many more tissues than previously determined, including human bone marrow and select regions of the human brain. In each of these tissues, we observed alternative splicing of various exons of PXR that generated multiple distinct PXR isoforms. The most abundant PXR alternative mRNA transcripts lacked 111 nucleotides, deleting 37 amino acids from the PXR LBD (PXR.2), or lacked 123 nt, deleting 41 amino acids from the PXR LBD (PXR.3). CYP3A4, a gene transcriptionally regulated by PXR, showed incomplete overlap with PXR in its tissue distribution. Quantitation of PXR mRNAs in human liver demonstrated that PXR.2 and PXR.3 represented 6.7% and 0.32% of total PXR mRNA transcripts. Brain expression of PXR prompted analysis of whether some brain acting chemicals were PXR ligands. The neurosteroids allopregnanolone and pregnanolone activated PXR and induced transcription of a CYP3A4-luciferase reporter. Nicotine, the psychoactive and addictive chemical in cigarettes, and a known inducer of brain CYP2B6, was an efficacious activator of PXR and inducer of CYP3A4 transcription. Because nicotine activation of PXR will enhance metabolism of nicotine to the non psychoactive cotinine, these results provide one molecular mechanism for the development of tolerance to nicotine. Moreover, the identification of PXR in many human tissues, such as brain, and activation by tissue specific ligands (such as neurosteroids) suggests additional biological roles for this receptor in these tissues. PMID- 15364542 TI - Environmental phthalate monoesters activate pregnane X receptor-mediated transcription. AB - Phthalate esters, widely used as plasticizers in the manufacture of products made of polyvinyl chloride, induce reproductive and developmental toxicities in rodents. The mechanism that underlies these effects of phthalate exposure, including the potential role of members of the nuclear receptor superfamily, is not known. The present study investigates the effects of phthalates on the pregnane X receptor (PXR), which mediates the induction of enzymes involved in steroid metabolism and xenobiotic detoxification. The ability of phthalate monoesters to activate PXR-mediated transcription was assayed in a HepG2 cell reporter assay following transfection with mouse PXR (mPXR), human PXR (hPXR), or the hPXR allelic variants V140M, D163G, and A370T. Mono-2-ethylhexyl phthalate (MEHP) increased the transcriptional activity of both mPXR and hPXR (5- and 15 fold, respectively) with EC50 values of 7-8 microM. mPXR and hPXR were also activated by monobenzyl phthalate (MBzP, up to 5- to 6-fold) but were unresponsive to monomethyl phthalate and mono-n-butyl phthalate (M(n)BP) at the highest concentrations tested (300 microM). hPXR-V140M and hPXR-A370T exhibited patterns of phthalate responses similar to the wild-type receptor. By contrast, hPXR-D163G was unresponsive to all phthalate monoesters tested. Further studies revealed that hPXR-D163G did respond to rifampicin, but required approximately 40 fold higher concentrations than wild-type receptor, suggesting that the ligand binding domain D163G variant has impaired ligand-binding activity. The responsiveness of PXR to activation by phthalate monoesters demonstrated here suggests that these ubiquitous environmental chemicals may, in part, exhibit their endocrine disruptor activities by altering PXR-regulated steroid hormone metabolism with potential adverse health effects in exposed individuals. PMID- 15364543 TI - Simultaneous, bidirectional inhibitory crosstalk between PPAR and STAT5b. AB - The transcription factors peroxisome proliferator-activated receptor (PPAR) and signal transducer and activator of transcription 5 (STAT5) activate genes involved in fatty acid metabolism (PPARalpha) and adipogenesis (PPARgamma) and mediate hormonal responses important for body growth, liver gene expression, and mammary gland development (STAT5a and STAT5b). These seemingly disparate pathways are subject to mutually inhibitory crosstalk, with growth hormone (GH)-activated STAT5 able to inhibit PPAR-regulated gene transcription by approximately 80%, and conversely, ligand-activated PPAR able to inhibit STAT5-regulated transcription to a similar degree. Given the co-expression of PPAR and STAT5 in multiple tissues, we investigated whether one of the factors dominates the inhibitory crosstalk. A PPAR-responsive Renilla luciferase reporter was constructed and used to monitor PPAR transcriptional activity in COS-1 cells co-transfected with a STAT5 firefly luciferase reporter. In cells co-stimulated with GH and a PPAR agonist, STAT5b inhibited expression of the PPAR-regulated Renilla luciferase reporter, whereas PPARalpha and PPARgamma inhibited transcription of the STAT5b regulated firefly luciferase reporter. The extent of the inhibitory crosstalk was dependent on the relative levels of expression of each transcription factor and on the relative concentrations of GH and PPAR agonist. Dose-response studies revealed that STAT5b was inhibited at an approximately 7-fold lower concentration of the PPARgamma ligand troglitazone than was required for activation of PPARgamma, indicating that only a portion of cellular PPARgamma is needed for STAT5b inhibition. Similarly, mono-(2-ethylhexyl)phthalate (MEHP), a reproductive toxicant and primary metabolite of the environmental chemical di-(2 ethylhexyl)phthalate (DEHP), inhibited STAT5b transcriptional activity with an EC50 value of 1.1 microM, corresponding to an approximately 10-fold lower concentration than required for activation of PPARgamma-dependent transcription. We conclude that the cross-inhibition between PPAR and STAT5 proceeds in a simultaneous, bidirectional manner. Exposure to phthalates and other environmental chemical activators of PPARs may thus lead to alteration of hormone induced, STAT5-regulated gene expression in tissues such as liver, fat and breast, where both transcription factors are expressed. Conversely, STAT5 activating hormones and cytokines may modulate the responsiveness of PPARs to their foreign chemical ligands. PMID- 15364544 TI - Regulation of the cytochrome P450 2A genes. AB - Cytochrome P450 monooxygenases of the CYP2A subfamily play important roles in xenobiotic disposition in the liver and in metabolic activation in extrahepatic tissues. Many of the CYP2A transcripts and enzymes are inducible by xenobiotic compounds, and the expression of at least some of the CYP2A genes is influenced by physiological status, such as circadian rhythm, and pathological conditions, such as inflammation, microbial infection, and tumorigenesis. Variability in the expression of the CYP2A genes, which differs by species, animal strain, gender, and organ, may alter the risks of chemical toxicity for numerous compounds that are CYP2A substrates. The mechanistic bases of these variabilities are generally not well understood. However, recent studies have yielded interesting findings in several areas, such as the role of nuclear factor 1 in the tissue-selective expression of CYP2A genes in the olfactory mucosa (OM); the roles of constitutive androstane receptor, pregnane X receptor (PXR), and possibly, peroxisome proliferator-activated receptors in transcriptional regulation of the Cyp2a5 gene; and the involvement of heterogeneous nuclear ribonucleoprotein A1 in pyrazole-induced stabilization of CYP2A5 mRNA. The aims of this minireview are to summarize current knowledge of the regulation of the CYP2A genes in rodents and humans, and to stimulate further mechanistic studies that will ultimately improve our ability to determine, and to understand, these variabilities in humans. PMID- 15364545 TI - Expression and characterization of human cytochrome P450 4F11: Putative role in the metabolism of therapeutic drugs and eicosanoids. AB - We previously reported the cDNA cloning of a new CYP4F isoform, CYP4F11. In the present study, we have expressed CYP4F11 in Saccharomyces cerevisiae and examined its catalytic properties towards endogenous eicosanoids as well as some clinically relevant drugs. CYP4F3A, also known as a leukotriene B4 omega hydroxylase, was expressed in parallel for comparative purposes. Our results show that CYP4F11 has a very different substrate profile than CYP4F3A. CYP4F3A metabolized leukotriene B4, lipoxins A4 and B4, and hydroxyeicosatetraenoic acids (HETEs) much more efficiently than CYP4F11. On the other hand, CYP4F11 was a better catalyst than CYP4F3A for many drugs such as erythromycin, benzphetamine, ethylmorphine, chlorpromazine, and imipramine. Erythromycin was the most efficient substrate for CYP4F11, with a Km of 125 microM and Vmax of 830 pmol min(-1) nmol(-1) P450. Structural homology modeling of the two proteins revealed some interesting differences in the substrate access channel including substrate recognition site 2 (SRS2). The model of CYP4F11 presents a more open access channel that may explain the ability to metabolize large molecules like erythromycin. Also, some wide variations in residue size, charge, and hydrophobicity in the FG loop region may contribute to differences in substrate specificity and activity between CYP4F3A and CYP4F11. PMID- 15364546 TI - Structural basis for the role in protein folding of conserved proline-rich regions in cytochromes P450. AB - The sequence PPGPXPXPXXGN is highly conserved in some cytochromes P450 across species from humans to plants. Within species, however, this specific sequence is not conserved although a proline-rich sequence is present. In CYP2C proteins, mutagenesis of the prolines and glycine in the PPGPXPXP part of the sequence results in less efficient assembly of native P450 molecules, but those molecules that are formed properly have specific activities similar to wild type suggesting that this region plays a role in folding of the protein. Further, the pattern of requirements for prolines was consistent with a left-handed polyproline II (PPII) helix structure in the PPGP segment. The recent determinations of the structures of CYP2C proteins permit a reinterpretation of these earlier experimental studies. In CYP2C5, the PPGPXPXP part of the sequence is in a left-handed polyproline II-like sequence with a 90 degrees bend at the glycine residue. The PXPXXGN part forms a hairpin structure with the remaining sequence protruding at a right angle. The structure forms tertiary interactions with protein segments centered on Tyr-61, Phe-219, and Tyr-376. van der Waals contacts of the rings of prolines with those of the equally highly conserved tyrosine residues may be particularly important. The proper positioning of the N-terminal segment containing Tyr-61 and the C-terminal segment containing Tyr-376 by interactions with the proline-rich region may be important for proper folding because these residues are in loops extending from four strands of a beta-sheet structure. A schematic model of the sequential folding interactions is presented, and although speculative, it is proposed that constraints required for the folding of a hydrophobic knob that will be inserted into the membrane contribute to the high conservation of the PPGPXPXPXXGN sequence in a subset of cytochromes P450. PMID- 15364547 TI - Mechanisms that regulate production of reactive oxygen species by cytochrome P450. AB - Mammalian cytochromes P450 (P450) are a family of heme-thiolate enzymes involved in the oxidative metabolism of a variety of endogenous and exogenous lipophilic compounds. Poor coupling of the P450 catalytic cycle results in continuous production of reactive oxygen species (ROS), which affects signaling pathways and other cellular functions. P450 generation of ROS is tightly controlled by regulation of gene transcription as well as by modulation of interactions between protein constituents of the monooxygenase that affects its activity, coupling, and stability. Malfunction of these mechanisms may result in a burst of ROS production, which can cause lipid peroxidation and oxidative stress. In turn, oxidative stress downregulates P450 levels by a variety of feedback mechanisms. This review provides an overview of recent advances in our understanding of these feedback mechanisms that serve to limit P450 production of ROS. Some of the more likely physiological and cellular effects of P450 generation of ROS are also discussed. PMID- 15364548 TI - Proteasome-dependent degradation of cytochromes P450 2E1 and 2B1 expressed in tetracycline-regulated HeLa cells. AB - The degradation of ethanol-inducible cytochrome P450 2E1 (CYP2E1) and phenobarbital-inducible cytochrome P450 2B1 (CYP2B1) expressed in tetracycline (Tc)-inducible HeLa cell lines was characterized. A steady-state pulse-chase analysis was used to determine a half-life of 3.8 h for CYP2E1 while the half life of CYP2B1 was 2.3-fold greater in the same cell line. In contrast, NADPH cytochrome P450 reductase which is constitutively expressed in Tc-HeLa cells had a half-life of about 30 h. Lactacystin and other selective proteasome inhibitors including N-benzyloxycarbonyl-leucyl-leucyl-leucinal (MG132) and N benzyloxycarbonyl-L-leucyl-L-leucyl-L-norvalinal (MG115) significantly inhibited both CYP2E1 and CYP2B1 degradation. The turnover of CYP2E1 was slightly inhibited by calpain inhibitors while CYP2B1 turnover was not altered. Inhibitors of lysosomal proteolysis had no effect on the degradation of either protein. Treatment of cells with brefeldin A did not alter the degradation of either P450 which suggested the degradation occurred in the endoplasmic reticulum (ER). Even in the presence of proteasome inhibitors high molecular weight ubiquitin conjugates were not observed. Mutagenesis of two putative ubiquitination sites (Lys 317 and 324) did not alter the degradation of CYP2E1. The role of ubiquitination in the degradation of CYP2E1 was also examined in a Chinese hamster mutant cell line E36ts20 that contains a thermolabile ubiquitin activating enzyme (E1). The turnover of CYP2E1 was not significantly different at the nonpermissive temperature in the ts20 when compared to the control E36 cells. Furthermore, the addition of the hsp90 inhibitors geldanamycin, herbimycin, and radicicol had no effect on the turnover of CYP2E1, differentiating the degradation of CYP2E1 from other substrates for proteasome-dependent degradation. PMID- 15364549 TI - Regulation of NAD(P)H:quininone oxidoreductase by glucocorticoids. AB - Previous studies in neonatal and adolescent rats as well as adrenalectomized rats have demonstrated that glucocorticoids regulate the expression of the rat NAD(P)H:quinone oxidoreductase gene (QOR). We used primary cultures of rat adult hepatocytes to document that added glucorticoids repress both the basal and 1,2 benzanthracene-induced expression of QOR mRNA by 65-70%. QOR enzyme activity and protein were concomitantly suppressed as well. The monotonic concentration response for repression of QOR gene products up to 100 microM DEX concentration demonstrated that the glucocorticoid receptor (GR) was most likely involved in this process. The lack of effect at higher concentration rules out a role for the Pregnane X receptor in this regulation by DEX. In addition, the anti-glucorticoid RU38486 blocked this negative regulation and the protein synthesis inhibitor cycloheximide had no effect on this repression process. Similar results of GR dependence were observed using a luciferase reporter construct containing the 5' flanking region of the human QOR gene using HepG2 cells. Collectively, these results demonstrate that GR must directly participate in the negative regulation of QOR gene expression by dexamethasone and other glucocorticoids in vivo. PMID- 15364550 TI - Regulation of UDP glucuronosyltransferases in the gastrointestinal tract. AB - The UDP glucuronosyltransferases (UGT) of the gastrointestinal (GI) tract have a crucial role in protection against the toxic effects of lipophilic chemicals in the environment. UGTs such as UGT1A7, UGT1A8, and UGT1A10 are exclusively expressed in gastrointestinal tissues, each with a unique tissue distribution pattern that is subject to interindividual variation. The factors regulating this tissue-specific expression and that contribute to variability are beginning to be elucidated. Studies on the UGT1A7, 1A8, 1A9, and 1A10 gene promoters in Caco-2 cells, an in vitro model of enterocytes of the gastrointestinal tract, have identified the caudal homeodomain transcription factor, Cdx2, as an important regulator of the UGT1A8 and 1A10 gene proximal promoters. This transcription factor is found exclusively in the small intestine and colon: it is absent in the gastric epithelium and the esophagus. Cdx2 regulates the UGT1A8 and 1A10 promoters in cooperation with hepatocyte nuclear factor 1alpha (HNF1alpha). It is noteworthy that UGT1A7 is not expressed in gastrointestinal tissue distal to the gastric mucosa and does not contain a Cdx2 binding site in its proximal promoter. Transcription factors, including Sp1, which differentially bind to the initiator regions of the UGT1A8, 1A9, and 1A10 promoters, also contribute to the differences in expression of these UGTs in Caco-2 cells. The identification of important regulatory regions of UGT genes expressed in the gastrointestinal tract, and the transcription factors that bind to these regions, will aid in the elucidation of factors that contribute to interindividual differences in gastrointestinal UGT expression. In turn, this will lead to further understanding of interindividual variation in the capacity of the GI tract to metabolize lipophilic chemicals and to act as a barrier to dietary toxins and orally administered drugs. PMID- 15364551 TI - Advances in vascular imaging. AB - The improvement of vascular imaging has allowed the acquisition of vascular images with higher resolution while minimizing the risks and discomfort to patients. As imaging developments continue to progress, establishment of valid clinical-based evidence, before the application of each innovation, will assure maintenance of the current trend. Also, as the vascular surgeon adopts a more comprehensive approach in the care of vascular patients, a high-quality endovascular suite will provide an environment for integration of both traditional open and evolving endovascular procedures. PMID- 15364552 TI - Advances in stent technology and drug-eluting stents. AB - From an initially simple concept, vascular stents have evolved into highly specialized instruments, and are poised to develop into sophisticated drug delivery systems. Although primary research has mostly targeted the coronary circulation, newer generation stents, deliverable almost anywhere, retarding neointimal hyperplasia and thrombosis, and promoting endothelial overgrowth, will offer the next approximation of true vascular healing. This, in turn,promises to promote less invasive, safer, and finally durable endovascular solutions to current surgical problems. PMID- 15364553 TI - Thrombolysis and mechanical thrombectomy for arterial disease. AB - Thrombolytic therapy has been around for close to 30 years now,but its exact role in the treatment of acute and chronic arterial occlusive disease continues to be debated. Studies have produced varying and contradictory results. We are still not sure if thrombolysis has any true advantages over surgical thromboembolectomy,or which lytic agent is the best. Nonetheless, the technique still plays an important role in the treatment of arterial occlusions. PMID- 15364554 TI - The use of closure devices. AB - The use of arterial closure devices offers significant benefits over manual compression in achieving groin hemostasis following catheter-based procedures. Several currently available devices provide rapid puncture site closure with complication rates similar to that of manual compression. Closure devices allow for early times to ambulation and hospital discharge, and have a high degree of patient satisfaction. Their use may be of particular benefit inpatients that are anticoagulated. We believe that their use should be strongly considered in all patients following femoral artery catheterization. PMID- 15364555 TI - Carotid artery stenting. AB - Efficacy for carotid artery stenting (CAS) has not been confirmed by randomized clinical trial methodology in conventional risk patients. Although carotid endarterectomy (CEA) is considered the preferred method for carotid revascularization in the management of patients with symptomatic and asymptomatic extracranial carotid occlusive disease, comparisons between CAS and CEA are now underway. In North America, the CREST (Carotid Revascularization Endarterectomy versus Stent Trial) protocol is now completing its lead-in or credentialing phase as randomization of cases is initiated. In Europe, the CAVATAS (Carotid and Vertebral Artery Transluminal Angioplasty Study) and Stent Protected Angioplasty versus Carotid Endarterectomy trials are recruiting symptomatic patients for randomization between CEA and CAS. It is anticipated that these trials will publish definitive results within the next 1 to 3 years, and help guide the referral of patients for CAS and CEA in the future. PMID- 15364556 TI - Current status of thoracic aortic endograft repair. AB - The advent and success of endovascular repair of abdominal aneurysms had led to the development of catheter-based techniques to treat thoracic aortic pathology. Such diseases, including thoracic aortic aneurysms, acute and chronic type B dissections,penetrating aortic ulcers, and traumatic aortic transection, challenge surgeons to perform complex operative repairs in high-risk patients. The minimally invasive nature of thoracic endografting may provide an attractive alternative therapy especially in patients deemed unfit for thoracotomy. A worldwide review of thoracic endografting demonstrates encouraging short- and midterm outcomes with significant reductions in morbidity and early mortality.Long-term surveillance will be crucial to discover complications unique to thoracic endovascular interventions and to determine which patients are appropriate candidates for stent-graft therapy. PMID- 15364557 TI - Suprarenal stents and other advances in endovascular aneurysm repair. AB - The history of endovascular aneurysm repair has already passed through its phases of "endoexuberance" and "endoscepticism" and there is now a balanced and broad understanding of the technology,its limits and advantages. Current endovascular technique and stent-graft design is the refinement of the accumulated endovascular experience til now. It is important to make note of these technological features incorporated in current stent-grafts and the clinical experience that precipitated their introduction as the technology progresses and new applications are proposed. PMID- 15364558 TI - What is the significance of endoleaks and endotension. AB - Endovascular repair has been used over a decade as a treatment of abdominal aortic aneurysm, and has become a widely accepted treatment method with a low rate of perioperative complications. Endoleak, perigraft blood flow outside endograft but within aneurysmsac, has been intensively studied during the last 10 years of endovascular aneurysm repair (EVR). The natural history of aneurysms with endoleak and the true clinical significance of various types of endoleaks remains unclear. Type I/III endoleak has been found to be associated with aneurysm rupture, while the risk of rupture of aneurysms with type II endoleak and endotension appears very small. In endotension, the aneurysm sac remains pressurized, even if there is no evidence of an endoleak. Currently,it is accepted that type I/III endoleaks should be corrected, preferably by endovascular means, due to the risk of rupture. If endovascular repair is not possible, then open conversion should be considered. The risk of conversion should be weighed against the risk of aneurysm rupture. Treatment of type II endoleaks and endotension is more controversial. In those with aneurysm enlargement,secondary interventions are often performed. PMID- 15364559 TI - Current endovascular management of atherosclerotic renal artery stenosis. AB - The recent advances in stent technology and renal endovascular management have provided a technically reproducible method of percutaneously treating atherosclerotic renal artery stenosis (RAS). In many centers, this has resulted in endovascular management being the primary therapy for atherosclerotic RAS. Although still controversial, it appears that endovascular management of RAS by primay stent deployment provides better blood pressure control than that afforded by best medical management. The impact on renal function is less than that found for hypertension, but there is evidence to suggest that the use of protection devices and primary stenting may enhance renal function outcomes. Whether the ultimate benefit of enhanced survival follows remains an important question and should be the subject of future prospective studies. PMID- 15364560 TI - Subintimal angioplasty for infrainguinal occlusive disease. AB - Subintimal angioplasty is a relatively new endovascular approach for the treatment of patients with femoropoliteal or tibial artery occlusion. The procedure involves intentionally creating a false channel or dissection around an arterial occlusion, then reentering the true lumen distally. The procedure has a relatively low incidence of complications and a high rate of technical success.Short-term patency has been fair in some reports and poor in others. Long term results are not available. Aspirin and possibly anticoagulation or antiplatelet agents may improve technical success and short-term patency. Further studies with longer follow-up are required before it can be recommended as a potentially viable alternative to arterial bypass. PMID- 15364561 TI - Endovascular management of iliofemoral venous occlusive disease. AB - New developments in the management of both acute and chronic iliac vein occlusive disease offer exciting options for the treatment of this often debilitating condition. Percutaneous clot removal using thrombolysis, mechanical thrombectomy, or a combination of the two is fast becoming the treatment of choice for patients presenting with acute iliofemoral deep vein thrombosis. Recanalization of chronic iliac vein occlusions with balloon angioplasty and stenting relieves symptoms of extremity swelling and pain in the majority of treated patients. Existing data provide convincing proof of the efficacy of endovascular recanalization procedures, and upcoming prospective, controlled trials will further clarify the role of these techniques in the therapeutic armamentarium. PMID- 15364562 TI - New approaches for the treatment of varicose veins. AB - New, minimally invasive techniques for the treatment of varicose veins including radiofrequency ablation (RFA), endovenous laser therapy (EVLT), and transilluminated power phlebectomy (TIPP) represent effective and possibly superior alternatives to traditional saphenous vein stripping and stab avulsion of varicose veins. Further experience with these procedures will help to determine which ones will become the method of choice for treating this complex disease process. Some of these new techniques may not prove to be effective in the hands of all treating specialists. However,it is very likely that some of these techniques, such as foam sclerotherapy and RFA, will replace the procedures that we currently use today. PMID- 15364563 TI - Endovascular management of vascular disease: current technology and applications. PMID- 15364564 TI - SAM domains can utilize similar surfaces for the formation of polymers and closed oligomers. AB - The mitogen-activated protein kinase (MAPK) Byr2 and its activator Ste4 are involved in the mating pheromone response pathway of Schizosaccharomyces pombe and interact via their SAM domains. SAM domains can self-associate to form higher order structures, including dimers, polymers and closed oligomers. Ste4-SAM is adjacent to a trimeric leucine zipper domain and we have shown previously that the two domains together (Ste4-LZ-SAM) bind to a monomeric Byr2-SAM with high affinity (Kd approximately 20 nM), forming a 3:1 complex. Here, we map the surfaces of Byr2-SAM and Ste4-SAM that is involved the interaction. A set of 38 mutants of Byr2-SAM and 33 mutants of Ste4-SAM were prepared, covering most of the protein surfaces. These mutants were purified and screened for binding, yielding a map of residues that are required for binding and a complementary map of residues that are not required. We find that the interface maps to regions of the SAM domains that are known to be important for the formation of SAM polymers. These results indicate that SAM domains can create a variety of oligomeric architectures utilizing common binding surfaces. PMID- 15364565 TI - Reduced contact order and RNA folding rates. AB - We investigated the relationship between RNA structure and folding rates accounting for hierarchical structural formation. Folding rates of two-state folding proteins correlate well with relative contact order, a quantitative measure of the number and sequence distance between tertiary contacts. These proteins do not form stable structures prior to the rate-limiting step. In contrast, most secondary structures are stably formed prior to the rate-limiting step in RNA folding. Accordingly, we introduce "reduced contact order", a metric that reflects only the number of residues available to participate in the conformational search after the formation of secondary structure. Plotting the folding rates and the reduced contact order from ten different RNAs suggests that RNA folding can be divided into two classes. To examine this division, folding rates of circularly permutated isomers are compared for two RNAs, one from each class. Folding rates vary by tenfold for circularly permuted Bacillus subtilis RNase P RNA isomers, whereas folding rates vary by only 1.2-fold for circularly permuted catalytic domains. This difference is likely related to the dissimilar natures of their rate-limiting steps. PMID- 15364566 TI - Down-regulation of early sea urchin histone H2A gene relies on cis regulative sequences located in the 5' and 3' regions and including the enhancer blocker sns. AB - The tandem repeated sea urchin alpha-histone genes are developmentally regulated by gene-specific promoter elements. Coordinate transcription of the five genes begins after meiotic maturation of the oocyte, continues through cleavage, and reaches its maximum at morula stage, after which these genes are shut off and maintained in a silenced state for the life cycle of the animal. Although cis regulative sequences affecting the timing and the level of expression of these genes have been characterized, much less is known about the mechanism of their repression. Here we report the results of a functional analysis that allowed the identification of the sequence elements needed for the silencing of the alpha-H2A gene at gastrula stage. We found that important negative regulative sequences are located in the 462 bp sns 5 fragment located in the 3' region. Remarkably, sns 5 contains the sns enhancer blocking element and the most 3' H2A codons. In addition, we made the striking observation that inhibition of the anti-enhancer activity of sns, by titration of the binding proteins in microinjected embryos, also affected the capability of sns 5 to down-regulate transgene expression at gastrula stage. A further sequence element essential for repression of the H2A gene was identified upstream of the enhancer, in the 5' region, and contains four GAGA repeats. Altogether these findings suggest that down-regulation of the alpha H2A gene occurs by the functional interaction of the 5' and 3' cis sequence elements. These results demonstrate the involvement of a genomic insulator in the silencing of gene expression. PMID- 15364567 TI - Statistical analysis of the spatial distribution of operons in the transcriptional regulation network of Escherichia coli. AB - We have performed a statistical analysis of the spatial distribution of operons along the DNA in the transcriptional regulation network of Escherichia coli. The analysis reveals that pairs of operons that regulate each other and those that are co-regulated tend to lie much closer to one another than would be expected for a random network. Moreover, these pairs of operons tend to be transcribed in diverging directions. This spatial arrangement of operons allows the upstream regulatory domains to overlap and interfere with each other and our analysis also demonstrates the statistical significance of this motif of overlapping operons. Overlapping operons afford additional regulatory control, such as the correlated or anticorrelated expression of operons. We show by a mean-field analysis of a feed-forward loop that overlapping operons can drastically enhance the performance of gene regulatory networks. Our results suggest that regulatory control can provide a selective pressure that drives operons together in the course of evolution. PMID- 15364568 TI - Defined structural changes occur in a nucleosome upon Amt1 transcription factor binding. AB - Here, we study the binding of the transcription factor Amt1 to its recognition site near the dyad of a highly positioned nucleosome. We find that the DNA binding domain of Amt1 binds to nucleosomes with only threefold reduced affinity compared to free DNA. We show by fluorescence resonance energy transfer that factor binding at the nucleosomal dyad is accompanied by the partial dissociation of the DNA ends from the histone octamer surface; however, no dissociation or subtle rearrangements of histone subunits is observed. A poly(dA.dT) DNA sequence element adjacent to the transcription factor binding site appears to facilitate factor binding, but is not essential. The methods that we describe here characterize for the first time the subtle structural changes that occur upon transcription factor binding to nucleosomes, and demonstrate the ability of the nucleosome to structurally adapt in response to outside influences. PMID- 15364569 TI - The Co-chaperone Sba1 connects the ATPase reaction of Hsp90 to the progression of the chaperone cycle. AB - The molecular chaperone Hsp90 mediates the ATP-dependent activation of a large number of proteins involved in signal transduction. During this process, Hsp90 was found to associate transiently with several accessory factors, such as p23/Sba1, Hop/Sti1, and prolyl isomerases. It has been shown that ATP hydrolysis triggers conformational changes within Hsp90, which in turn are thought to mediate conformational changes in the substrate proteins, thereby causing their activation. The specific role of the partner proteins in this process is unknown. Using proteins from Saccharomyces cerevisiae, we characterized the interaction of Hsp90 with its partner protein p23/Sba1. Our results show that the nucleotide dependent N-terminal dimerization of Hsp90 is necessary for the binding of Sba1 to Hsp90 with an affinity in the nanomolar range. Two Sba1 molecules were found to bind per Hsp90 dimer. Sba1 binding to Hsp90 resulted in a decreased ATPase activity, presumably by trapping the hydrolysis state of Hsp90ATP. Ternary complexes of Hsp90Sba1 could be formed with the prolyl isomerase Cpr6, but not with Sti1. Based on these findings, we propose a model that correlates the ordered assembly of the Hsp90 co-chaperones with distinct steps of the ATP hydrolysis reaction during the chaperone cycle. PMID- 15364570 TI - Positive selection of synonymous mutations in vesicular stomatitis virus. AB - Prevailing evolutionary forces are typically deduced from the pattern of differences in synonymous and non-synonymous mutations, under the assumption of neutrality in the absence of amino acid change. We determined the complete sequence of ten vesicular stomatitis virus populations evolving under positive selection. A significant number of the mutations occurred independently in two or more strains, a process known as parallel evolution, and a substantial fraction of the parallel mutations were silent. Parallel evolution was also identified in non-coding regions. These results indicate that silent mutations can significantly contribute to adaptation in RNA viruses, and relative frequencies of synonymous and non-synonymous substitutions may not be useful to resolve their evolutionary history. PMID- 15364571 TI - Molecular basis of fitness loss and fitness recovery in vesicular stomatitis virus. AB - Viral populations subjected to repeated genetic bottleneck accumulate deleterious mutations in a process known as Muller's ratchet. Asexual viruses, such as vesicular stomatitis virus (VSV) can recover from Muller's ratchet by replication with large effective population sizes. However, mutants with a history of bottleneck transmissions often show decreased adaptability when compared to non bottlenecked populations. We have generated a collection of bottlenecked mutants and allowed them to recover by large population passages. We have characterized fitness changes and the complete genomes of these strains. Mutations accumulated during the operation of Muller's ratchet led to the identification of two potential mutational hot spots in the VSV genome. As in other viral systems, transitions were more common than transversions. Both back mutation and compensatory mutations contributed to recovery, although a significant level of fitness increase was observed in nine of the 13 bottlenecked strains with no obvious changes in the consensus sequence. Additional replication of three strains resulted in the fixation of single point mutations. Only two mutations previously found in non-bottlenecked, high-fitness populations that had been adapting to the same environment were identified in the recovered strains. PMID- 15364572 TI - Monovalent ion-mediated folding of the Tetrahymena thermophila ribozyme. AB - The time-course of monovalent cation-induced folding of the L-21 Sca1 Tetrahymena thermophila ribozyme and a selected mutant was quantitatively followed using synchrotron X-ray (.OH) footprinting. Initiating folding by increasing the concentration of either Na+ or K+ to 1.5M from an initial condition of approximately 0.008 M Na+ at 42 degrees C resulted in the complete formation of tertiary contacts within the P5abc subdomain and between the peripheral helices within the dead time of our measurements (k>50 s(-1)). These results contrast with folding rates of 2-0.2 s(-1) previously observed for formation of these contacts in 10mM Mg2+ from the same initial condition. Thus, the initial formation of native tertiary contacts is inhibited by divalent but not monovalent cations. The native contacts within the catalytic core form without a detectable burst phase at rates of 0.4-1.0 s(-1) in a manner reminiscent of the Mg2+ dependent folding behavior, although tenfold faster. The tertiary interactions stabilizing the catalytic core interaction with P4-P6 and P2.1, as well as one of the protections internal for the P4-P6 domain, display progress curves with appreciable burst amplitudes and a phase comparable in rate to that of the catalytic core. That the slow folding of the ribozyme's core is a consequence of the alt-P3 secondary structure is shown by the 100% burst phase amplitudes that are observed for folding of the U273A mutant ribozyme within which the native secondary structure (P3) is strengthened. Thus, formation of a misfolded intermediate(s) resulting from the alt-P3 secondary structure is independent of ion valency while the rate at which the respective intermediates are resolved is sensitive to ion valency. The overall portrait painted by these results is that ion valency differentially affects steps in the folding process and that folding in monovalent ion alone for the U273A mutant Tetrahymena ribozyme is fast and direct. PMID- 15364573 TI - Intermediate DNA repair activity associated with the 322delG allele of the fanconi anemia complementation group C gene. AB - Fanconi anemia (FA) is an autosomal recessive disorder associated with pancytopenia and cancer susceptibility. The disorder is heterogeneous, with at least nine complementation groups having been identified. Several recent studies have suggested that defective plasmid DNA end-joining is a consistent feature of FA cells. It was therefore surprising to discover a strain of fibroblasts from an FA patient that possessed wild-type plasmid DNA end-joining activity. Unlike other FA strains, these fibroblasts have wild-type levels of homologous DNA recombination activity and are relatively insensitive to restriction endonuclease induced death. Interestingly, while end-joining in a number of FA fibroblast strains belonging to complementation groups A, C, and D2 was approximately 70% precise, end-joining in this latter strain of fibroblasts was more than 95% imprecise. Analysis revealed that these latter cells harbored an allele of the FA C gene, referred to as 322delG, that encodes an amino-terminal truncated protein. The relative rarity of this allele precluded the analysis of other FA fibroblast strains; however, studies revealed that overexpression of this allele in normal cells recapitulated the DNA end-joining phenotype seen in the 322delG FA fibroblast strain. These results indicate that DNA end-joining in fibroblasts expressing the 322delG allele of the FA-C gene in fibroblasts is highly imprecise; however, the DNA repair efficiency of these cells is more normal than that commonly associated with FA fibroblasts. This conclusion is intriguing, since a number of reports have suggested that patients harboring this allele exhibit a milder clinical course than do individuals with other alleles of the FA C gene. PMID- 15364574 TI - Dual functions, clamp opening and primer-template recognition, define a key clamp loader subunit. AB - Clamp loader proteins catalyze assembly of circular sliding clamps on DNA to enable processive DNA replication. During the reaction, the clamp loader binds primer-template DNA and positions it in the center of a clamp to form a topological link between the two. Clamp loaders are multi-protein complexes, such as the five protein Escherichia coli, Saccharomyces cerevisiae, and human clamp loaders, and the two protein Pyrococcus furiosus and Methanobacterium thermoautotrophicum clamp loaders, and thus far the site(s) responsible for binding and selecting primer-template DNA as the target for clamp assembly remain unknown. To address this issue, we analyzed the interaction between the E.coli gamma complex clamp loader and DNA using UV-induced protein-DNA cross-linking and mass spectrometry. The results show that the delta subunit in the gamma complex makes close contact with the primer-template junction. Tryptophan 279 in the delta C-terminal domain lies near the 3'-OH primer end and may play a key role in primer-template recognition. Previous studies have shown that delta also binds and opens the beta clamp (hydrophobic residues in the N-terminal domain of delta contact beta. The clamp-binding and DNA-binding sites on delta appear positioned for facile entry of primer-template into the center of the clamp and exit of the template strand from the complex. A similar analysis of the S.cerevisiae RFC complex suggests that the dual functionality observed for delta in the gamma complex may be true also for clamp loaders from other organisms. PMID- 15364575 TI - Crystal structures of Escherichia coli RecA in a compressed helical filament. AB - The X-ray crystal structure of uncomplexed Escherichia coli RecA protein has been determined in three new crystal forms at resolutions of 1.9 A, 2.0 A, and 2.6 A. The RecA protein used for this study contains the extra residues Gly-Ser-His-Met at the N terminus, but retains normal ssDNA-dependent ATPase and coprotease activities. In all three crystals, RecA is packed in a right-handed helical filament with a pitch of approximately 74 A. These RecA filaments are compressed relative to the original crystal structure of RecA, which has a helical pitch of 82.7 A. In the structures of the compressed RecA filament, the monomer-monomer interface and the core domain are essentially the same as in the RecA structure with the 83 A pitch. The change in helical pitch is accommodated by a small movement of the N-terminal domain, which is reoriented to preserve the contacts it makes at the monomer-monomer interface. The new crystal structures show significant variation in the orientation and conformation of the C-terminal domain, as well as in the inter-filament packing interactions. In crystal form 2, a calcium ion is bound closely to a beta-hairpin of the C-terminal domain and to Asp38 of a neighboring filament, and residues 329-331 of the C-terminal tail become ordered to contact a neighboring filament. In crystal forms 3 and 4, a sulfate ion or a phosphate anion is bound to the same site on RecA as the beta phosphate group of ADP, causing an opening of the P-loop. Altogether, the structures show the conformational variability of RecA protein in the crystalline state, providing insight into many aspects of RecA function. PMID- 15364576 TI - Distinguishing structural and functional restraints in evolution in order to identify interaction sites. AB - Structural genomics projects are producing many three-dimensional structures of proteins that have been identified only from their gene sequences. It is therefore important to develop computational methods that will predict sites involved in productive intermolecular interactions that might give clues about functions. Techniques based on evolutionary conservation of amino acids have the advantage over physiochemical methods in that they are more general. However, the majority of techniques neither use all available structural and sequence information, nor are able to distinguish between evolutionary restraints that arise from the need to maintain structure and those that arise from function. Three methods to identify evolutionary restraints on protein sequence and structure are described here. The first identifies those residues that have a higher degree of conservation than expected: this is achieved by comparing for each amino acid position the sequence conservation observed in the homologous family of proteins with the degree of conservation predicted on the basis of amino acid type and local environment. The second uses information theory to identify those positions where environment-specific substitution tables make poor predictions of the overall amino acid substitution pattern. The third method identifies those residues that have highly conserved positions when three dimensional structures of proteins in a homologous family are superposed. The scores derived from these methods are mapped onto the protein three-dimensional structures and contoured, allowing identification clusters of residues with strong evolutionary restraints that are sites of interaction in proteins involved in a variety of functions. Our method differs from other published techniques by making use of structural information to identify restraints that arise from the structure of the protein and differentiating these restraints from others that derive from intermolecular interactions that mediate functions in the whole organism. PMID- 15364577 TI - Crystal complex structures reveal how substrate is bound in the -4 to the +2 binding sites of Humicola grisea Cel12A. AB - As part of an ongoing enzyme discovery program to investigate the properties and catalytic mechanism of glycoside hydrolase family 12 (GH 12) endoglucanases, a GH family that contains several cellulases that are of interest in industrial applications, we have solved four new crystal structures of wild-type Humicola grisea Cel12A in complexes formed by soaking with cellobiose, cellotetraose, cellopentaose, and a thio-linked cellotetraose derivative (G2SG2). These complex structures allow mapping of the non-covalent interactions between the enzyme and the glucosyl chain bound in subsites -4 to +2 of the enzyme, and shed light on the mechanism and function of GH 12 cellulases. The unhydrolysed cellopentaose and the G2SG2 cello-oligomers span the active site of the catalytically active H.grisea Cel12A enzyme, with the pyranoside bound in subsite -1 displaying a S31 skew boat conformation. After soaking in cellotetraose, the cello-oligomer that is found bound in site -4 to -1 contains a beta-1,3-linkage between the two cellobiose units in the oligomer, which is believed to have been formed by a transglycosylation reaction that has occurred during the ligand soak of the protein crystals. The close fit of this ligand and the binding sites occupied suggest a novel mixed beta-glucanase activity for this enzyme. PMID- 15364578 TI - The structure of Rigidoporus lignosus Laccase containing a full complement of copper ions, reveals an asymmetrical arrangement for the T3 copper pair. AB - Laccase is a multicopper blue oxidase that couples the four-electron reduction of oxygen with the oxidation of a broad range of organic substrates, including phenols and arylamines. The enzyme is the object of intense biotechnological research, due to its employment in bioremediation of soils and water as well as in other biotechnological applications. We report here the cDNA and protein sequences, the post-translational modifications, the crystallization and X-ray structure determination of a laccase from the white-rot fungus Rigidoporus lignosus. The amino acid residues sequence deduced from cDNA clearly identified a pre-sequence of 21 residues representing the signal for extra-cellular localization. Mass spectrometry analysis performed on the salvage enzyme, confirmed the deduced sequence and precisely mapped two glycosylation sites at Asn337 and Asn435, determining the nature of the bound glycosidic moieties. The crystal structure was determined at 1.7A resolution from perfectly hemihedrally twinned crystals, by molecular replacement technique. While the overall structure closely resembled those reported for other fungal laccases, the analysis of the T2/T3 trinuclear cluster revealed an unprecedented coordination sphere for the T3 copper pair. No bridging oxygen ligand was present between the two T3 copper ions, which were no longer symmetrically coordinated. The observed structure could represent an intermediate along the process of four-electron reduction of oxygen to water taking place at the trinuclear copper cluster. PMID- 15364579 TI - The structure of MBL-associated serine protease-2 reveals that identical substrate specificities of C1s and MASP-2 are realized through different sets of enzyme-substrate interactions. AB - A family of serine proteases mediates the proteolytic cascades of several defense mechanisms in vertebrates, such as the complement system, blood coagulation and fibrinolysis. These proteases usually form large complexes with other glycoproteins. Their common features are their modular structures and restricted substrate specificities. The lectin pathway of complement, where mannose-binding lectin (MBL) recognizes the carbohydrate structures on pathogens, is activated by mannose-binding lectin-associated serine protease-2 (MASP-2). We present the 2.25A resolution structure of the catalytic fragment of MASP-2 encompassing the second complement control protein module (CCP2) and the serine protease (SP) domain. The CCP2 module stabilizes the structure of the SP domain as demonstrated by differential scanning calorimetry measurements. The asymmetric unit contains two molecules with different CCP-SP domain orientations, reflecting increased modular flexibility at the CCP2/SP joint. This flexibility may partly explain the ability of the MASP-2 dimer to perform all of its functions alone, whereas the same functions are mediated by the much larger C1r2-C1s2 tetramer in the C1 complex of the classical pathway. The main scaffold of the MASP-2 SP domain is chymotrypsin-like. Eight surface loops determine the S1 and other subsite specificities. Surprisingly, some surface loops of MASP-2, e.g. loop 1 and loop 2, which form the S1 pocket are similar to those of trypsin, and show significant differences if compared with those of C1s, indicating that the nearly identical substrate specificities of C1s and MASP-2 are realized through different sets of enzyme-substrate interactions. PMID- 15364580 TI - Inter-domain motions of the N-domain of the KdpFABC complex, a P-type ATPase, are not driven by ATP-induced conformational changes. AB - P-type ATPases are involved in the active transport of ions across biological membranes. The KdpFABC complex (P-type ATPase) of Escherichia coli is a high affinity K+ uptake system that operates only when the cell experiences osmotic stress or K+ limitation. Here, we present the solution structure of the nucleotide binding domain of KdpB (backbone RMSD 0.17 A) and a model of the AMP PNP binding mode based on intermolecular distance restraints. The calculated AMP PNP binding mode shows the purine ring of the nucleotide to be "clipped" into the binding pocket via a pi-pi-interaction to F377 on one side and a cation-pi interaction to K395 on the other. This binding mechanism seems to be conserved in all P-type ATPases, except the heavy metal transporting ATPases (type IB). Thus, we conclude that the Kdp-ATPase (currently type IA) is misgrouped and has more similarities to type III ATPases. The KdpB N-domain is the smallest and simplest known for a P-type ATPase, and represents a minimal example of this functional unit. No evidence of significant conformational changes was observed within the N domain upon nucleotide binding, thus ruling out a role for ATP-induced conformational changes in the reaction cycle. PMID- 15364581 TI - Cofilin induced conformational changes in F-actin expose subdomain 2 to proteolysis. AB - Cofilin/ADF affects strongly the structure of actin filaments and especially the intermolecular contacts of the DNase I binding loop (D-loop) in subdomain 2. In G actin, the D-loop is cleaved by subtilisin between Met47 and Gly48, while in F actin this cleavage is inhibited. Here, we report that yeast cofilin, which is resistant to both subtilisin and trypsin, accelerates greatly the rate of subtilisin cleavage of this loop in F-actin at pH 6.8 and at pH 8.0. Similarly, cofilin accelerates strongly the tryptic cleavage in F-actin of loop 60-69 in subdomain 2, at Arg62 and Lys68. The acceleration of the loops' proteolysis cannot be attributed to an increased treadmilling of F-actin for the following reasons: (i) the rate of subtilisin cleavage is independent of pH between pH 6.8 and 8.0, unlike F-actin depolymerization, which is pH-dependent; (ii) at high concentrations of protease the cleavage rate of F-actin in the presence of cofilin is faster than the rate of monomer dissociation from the pointed end of TRC-labeled F-actin, which limits the rate of treadmilling; and (iii) cofilin also accelerates the rate of subtilisin cleavage of F-actin in which the treadmilling is blocked by interprotomer cross-linking of the D-loop to the C terminus on an adjacent protomer. This suggests a substantial flexibility of the D-loop in the cross-linked F-actin. The increased cleavage rates of the D-loop and loop 60-69 reveal extensive exposure of subdomain 2 in F-actin to proteolytic enzymes by cofilin. PMID- 15364582 TI - The structural basis of macrolide-ribosome binding assessed using mutagenesis of 23S rRNA positions 2058 and 2059. AB - Macrolides are a diverse group of antibiotics that inhibit bacterial growth by binding within the peptide tunnel of the 50S ribosomal subunit. There is good agreement about the architecture of the macrolide site from different crystallography studies of bacterial and archaeal 50S subunits. These structures show plainly that 23S rRNA nucleotides A2058 and A2059 are located accessibly on the surface of the tunnel wall where they act as key contact sites for macrolide binding. However, the molecular details of how macrolides fit into this site remain a matter of contention. Here, we have generated an isogenic set of single and dual substitutions at A2058 and A2059 in Mycobacterium smegmatis to investigate the effects of the rRNA mutations on macrolide binding. Resistances conferred to a comprehensive array of 11 macrolide compounds are used to assess models of macrolide binding predicted from the crystal structures. The data indicate that all macrolides and their derivatives bind at the same site in the tunnel with their C5 amino sugar in a similar orientation. Our data are compatible with the lactone rings of 14-membered and 16-membered macrolides adopting different conformations, enabling the latter compounds to avoid a steric clash with 2058G. This difference, together with interactions conveyed via substituents that are specific to certain ketolide and macrolide sub-classes, influences the binding to the large ribosomal subunit. Our genetic data show no support for a derivatized-macrolide binding site that has been proposed to be located further down the tunnel. PMID- 15364583 TI - The crystal structure of a complex of Campylobacter jejuni dUTPase with substrate analogue sheds light on the mechanism and suggests the "basic module" for dimeric d(C/U)TPases. AB - The crystal structure of the dUTPase from the important gastric pathogen Campylobacter jejuni has been solved at 1.65 A spacing. This essential bacterial enzyme is the second representative of the new family of dimeric dUTPases to be structurally characterised. Members of this family have a novel all-alpha fold and are unrelated to the all-beta dUTPases of the majority of organisms including eukaryotes such as humans, bacteria such as Escherichia coli, archaea like Methanococcus jannaschii and animal viruses. Therefore, dimeric dUTPases can be considered as candidate drug targets. The X-ray structure of the C.jejuni dUTPase in complex with the non-hydrolysable substrate analogue dUpNHp allows us to define the positions of three catalytically significant phosphate-binding magnesium ions and provides a starting point for a detailed understanding of the mechanism of dUTP/dUDP hydrolysis by dimeric dUTPases. Indeed, a water molecule present in the structure is ideally situated to act as the attacking nucleophile during hydrolysis. A comparison of the dUTPases from C.jejuni and Trypanosoma cruzi reveals a common fold with certain distinct features, both in the rigid and mobile domains as defined in the T.cruzi structure. Homologues of the C.jejuni dUTPase have been identified in several other bacteria and bacteriophages, including the dCTPase of phage T4. Sequence comparisons of these proteins define a new superfamily of d(C/U)TPases that includes three distinct enzyme families: (1) dUTPases in trypanosomatides, C.jejuni and several other Gram-negative bacteria, (2) predicted dUTPases in various Gram-positive bacteria and their phages, and (3) dCTP/dUTPases in enterobacterial T4-like phages. All these enzymes share a basic module that consists of two alpha-helices from the rigid domain, two helices from the mobile domain and connecting loops. These results in concert with a number of conserved residues responsible for interdomain cross talk provide valuable insight towards rational drug design. PMID- 15364584 TI - Gated electron transfers and electron pathways in azurin: a NMR dynamic study at multiple fields and temperatures. AB - Dynamic properties of electron transfer pathways in a small blue copper cupredoxin are explored using an extensive 15N NMR relaxation study of reduced Pseudomonas aeruginosa azurin at four magnetic fields (500-900 MHz) and at two temperatures chosen well below the melting point of the protein. Following a careful model-free analysis, several protein regions with different dynamic regimes are identified. Nanosecond time-scale mobility characterizes various residues of the hydrophobic surface patch believed to mark the natural entry point for electrons, notably the surface-exposed copper-ligand His117. These findings are consistent with a gated electron transfer process according to the "dynamic docking" model. Residues 47-49 along intramolecular pathways of electrons show rigidity that is remarkably conserved when increasing the temperature. Three different conformational exchange processes were observed in the millisecond range, one near the only disulfide bridge in the molecule and two near the copper ion. The latter two processes are consistent with previous data such as crystal structures at various pH values and NMR relaxation dispersion experiments; they may indicate an additional gated electron transfer mechanism at slower time-scales. PMID- 15364585 TI - Structural insights into the neuroprotective-acting carbonyl reductase Sniffer of Drosophila melanogaster. AB - In vivo studies with the fruit-fly Drosophila melanogaster have shown that the Sniffer protein prevents age-dependent and oxidative stress-induced neurodegenerative processes. Sniffer is a NADPH-dependent carbonyl reductase belonging to the enzyme family of short-chain dehydrogenases/reductases (SDRs). The crystal structure of the homodimeric Sniffer protein from Drosophila melanogaster in complex with NADP+ has been determined by multiple-wavelength anomalous dispersion and refined to a resolution of 1.75 A. The observed fold represents a typical dinucleotide-binding domain as detected for other SDRs. With respect to the cofactor-binding site and the region referred to as substrate binding loop, the Sniffer protein shows a striking similarity to the porcine carbonyl reductase (PTCR). This loop, in both Sniffer and PTCR, is substantially shortened compared to other SDRs. In most enzymes of the SDR family this loop adopts a well-defined conformation only after substrate binding and remains disordered in the absence of any bound ligands or even if only the dinucleotide cofactor is bound. In the structure of the Sniffer protein, however, the conformation of this loop is well defined, although no substrate is present. Molecular modeling studies provide an idea of how binding of substrate molecules to Sniffer could possibly occur. PMID- 15364586 TI - The disintegrin echistatin stabilizes integrin alphaIIbbeta3's open conformation and promotes its oligomerization. AB - We have employed echistatin, a 5.4 kDa snake venom disintegrin, as a model protein to investigate the paradox that small ligand-mimetics can bind to the resting alphaIIbbeta3 integrin while adhesive macromolecules cannot. We characterized the interactions between purified human alphaIIbbeta3 and two recombinant echistatin variants: rEch (1-49) M28L, chosen for its selectivity toward beta3-integrins, and rEch (1-40) M28L, a carboxy-terminal truncation mutant. While both contain an RGD integrin targeting sequence, only rEch (1-49) M28L was an effective inhibitor of alphaIIbbeta3 function. Electron microscopy of rotary shadowed specimens yielded a variety of alphaIIbbeta3 conformers ranging from compact, spherical particles (maximum dimension 22 nm) to the classical "head with two tails" forms (32 nm). The population of larger particles (42-56 nm) increased from 17% to 28% in the presence of rEch (1-49) M28L, indicative of ligand-induced oligomerization. Sedimentation velocity measurements demonstrated that both full length and truncated echistatin perturbed alphaIIbbeta3's solution structure, yielding slower-sedimenting open conformers. Dynamic light scattering showed that rEch (1-49) M28L protected alphaIIbbeta3 from thermal aggregation, raising its transition mid-point from 46 degrees C to 69 degrees C; a smaller shift resulted with rEch (1-40) M28L. Sedimentation equilibrium demonstrated that both echistatin ligands induced substantial alphaIIbbeta3 dimerization. van't Hoff analysis revealed a pattern of entropy/enthalpy compensation similar to tirofiban, a small RGD ligand-mimetic that binds tightly to alphaIIbbeta3, but yields smaller conformational perturbations than echistatin. We propose that echistatin may serve as a paradigm for understanding multidomain adhesive macromolecules because its ability to modulate alphaIIbbeta3's structure resides on an RGD loop, while full disintegrin activity requires an auxiliary site that includes the carboxy-terminal nine amino acid residues. PMID- 15364587 TI - Molecular structure of the rod domain of dictyostelium filamin. AB - Dictyostelium discoideum filamin (ddFLN) is a two-chain F-actin crosslinking protein with an N-terminal actin-binding domain and a rod domain constructed from six tandem repeats of a 100 residue motif that has an immunoglobulin (Ig) fold. We report the 2.8 A resolution crystal structure of a homodimer of rod repeats 4, 5 and 6. The two chains are arranged in an antiparallel fashion and form an elongated element, which is shortened, however, compared to a fully extended, linear configuration because the long axis of each Ig domain is arranged at an angle to the long axis of the rod. Same arrangement of repeats should also be present in the rod domain of human FLNa, much longer than Dictyostelium FLN, which forms an extended structure able to crosslink F-actin chains over distances of more than 1000 A. PMID- 15364588 TI - The efficiency of mispaired ligations by lambda integrase is extremely sensitive to context. AB - The integrase protein (Int) of phage lambda is a well-studied representative of the tyrosine recombinase family, whose defining features are two sequential pairs of DNA cleavage/ligation reactions that proceed via a 3' phosphotyrosine covalent intermediate to first form and then resolve a Holliday junction recombination intermediate. We devised an assay that takes advantage of DNA hairpin formation at one Int target site to trap Int cleavages at a different target site, and thereby reveal iterative cycles of cleavage and ligation that would otherwise be undetected. Using this assay and others to compare wild-type Int and a mutant (R169D) defective in forming proper dimer/tetramer interfaces, we found that the efficiency of "bottom-strand" DNA cleavage by wild-type Int, but not R169D, is very sensitive to the base-pair at the "top-strand" cleavage site, seven base pairs away. We show that this is related to the finding that hairpin formation involving ligation of a mispaired base is much faster for R169D than for wild type Int, but only in the context of a multimeric complex. During resolution of Holliday junction recombination intermediates, wild-type Int, but not R169D, is very sensitive to homology at the sites of ligation. A long-sought insight from these results is that during Holliday junction resolution the tetrameric Int complex remains intact until after ligation of the product helices has been completed. This contrasts with models in which the second pair of DNA cleavages is a trigger for dissolution of the recombination complex. PMID- 15364589 TI - Structural delineation of the calcineurin-NFAT interaction and its parallels to PP1 targeting interactions. AB - Calcineurin is a phosphoprotein phosphatase that channels intracellular Ca signals into multiple biological pathways. Calcineurin is known to interact directly with its substrate nuclear factor of activated T cells (NFAT or NFATc), with other substrates, and with several targeting and scaffold proteins including AKAP79 and Cabin1/cain. The calcineurin-NFAT interaction depends on recognition of a PxIxIT sequence motif present in NFAT-family proteins and in certain other calcineurin-interacting proteins. Here, we define the structural basis for the interaction of calcineurin with NFAT and with other proteins possessing the PxIxIT motif. The calcineurin-PxIxIT contact has a direct parallel in the contact of protein phosphatase 1 with its regulatory proteins, suggesting that the evolution of these related phosphatases involved local remodelling of an ancestral docking site. PMID- 15364590 TI - Mercury in canned tuna: white versus light and temporal variation. AB - There are abundant data and advisories for mercury levels in wild fish, but far fewer for commercial fish that compose a large majority of the fish most people eat. Until recently, relatively little attention has been devoted to examining mercury in canned tuna, despite its great importance in human diets. There is substantial media coverage of the benefits and risk from fish consumption, but few peer-reviewed data on canned tuna, the most commonly consumed fish in the United States. In this paper, we examine the levels of total mercury in canned tuna obtained from a New Jersey grocery store from 1998 to 2003, looking for temporal consistency within this data set and particularly for comparison with the Food and Drug Administration's 1991 study. We analyzed 168 cans individually for total mercury. All values are reported as parts per million (= microg/g) on a wet weight basis. In a subset of samples analyzed for total and inorganic mercury, the inorganic mercury was below detection levels; hence at least 89% of the mercury can be considered methylmercury. We found that white-style tuna had significantly more total mercury (mean 0.407 ppm) than light-style tuna (mean 0.118 ppm), presumably reflecting that "white" tuna is albacore, a species relatively larger than the skipjack tuna, which is commonly available as "light" or "chunk light." The maximum mercury in a can was 0.997 ppm, but 25% of white tuna samples exceeded 0.5 ppm. Data suggest a slight increase in levels since 1991, and mercury levels were significantly higher in 2001 than in other years. The mean level of mercury in white tuna (mean 0.407 ppm) was significantly higher than the mean value of 0.17 ppm currently used by the U.S. Food and Drug Administration (FDA) in its risk assessment and public information. There were no significant differences in mercury levels in tuna packed in oil compared to water. Draining contents had no effect on mercury levels, and the fluid, both oil and water, contained little mercury. These data indicate that people who eat canned tuna frequently can choose light tuna and reduce their mercury intake. Canned mackerel had much lower levels of mercury than tuna. Since cans of white tuna frequently exceed the FDA's original action level of 0.5 ppm, it would be prudent to continue some systematic monitoring of the nation's canned fish supply, particularly as the targets of commercial fisheries inevitably change as certain stocks become depleted. PMID- 15364591 TI - Amoebae and other protozoa in material samples from moisture-damaged buildings. AB - Mold growth in buildings has been shown to be associated with adverse health effects. The fungal and bacterial growth on moistened building materials has been studied, but little attention has been paid to the other organisms spawning in the damaged materials. We examined moist building materials for protozoa, concentrating on amoebae. Material samples (n = 124) from moisture-damaged buildings were analyzed for amoebae, fungi, and bacteria. Amoebae were detected in 22% of the samples, and they were found to favor cooccurrence with bacteria and the fungi Acremonium spp., Aspergillus versicolor, Chaetomium spp., and Trichoderma spp. In addition, 11 seriously damaged samples were screened for other protozoa. Ciliates and flagellates were found in almost every sample analyzed. Amoebae are known to host pathogenic bacteria, such as chlamydiae, legionellae, and mycobacteria and they may have a role in the complex of exposure that contributes to the health effects associated with moisture damage in buildings. PMID- 15364592 TI - Hair mercury levels versus freshwater fish consumption in household members of Swedish angling societies. AB - Hair mercury levels were determined in 143 individuals from households of members in angling societies in an area of Sweden with many lakes that have freshwater fish with relatively high mercury levels. Thus, the individuals had a potentially high intake of methyl mercury. The mean mercury concentration of pike and perch was approximately 0.7 microg/g. One-third of the subjects consumed these freshwater fish at least once a week. As could be expected, there was a clear increase in hair Hg with reported freshwater fish consumption (P < 0.001). The median mercury level in hair was 0.9 microg Hg/g for the whole group, and for those who reported consumption of freshwater fish at least once a week it was 1.8 microg/g. The highest hair mercury level was 18.5 microg/g, in a man who consumed pike and perch several times per week. Men had higher hair Hg than women, also when stratified for fish consumption. This was verified in 32 couples, of which the man and woman consumed the same fish and reported the same consumption. The median hair mercury level in these 32 couples was 1.3 microg/g for men and 0.8 microg/g for women (P = 0.002). About half of the subjects had hair mercury exceeding 1 microg/g, corresponding to the reference dose (RfD) of 0.1 microg of mercury per kilogram body weight set by the US Environmental Protection Agency. Although the RfD applies to all populations, the most at-risk group at these levels is pregnant women. There were only 2 women (of 12) of fertile age with hair mercury above 1 microg/g. In Sweden pregnant women are advised not to eat perch and pike at all during pregnancy. Since fish is rich in many important nutrients, it is unsatisfactory that fish consumption must be restricted, and thus there is a need to reduce mercury levels in fish. PMID- 15364593 TI - Relationship between blood lead concentration and dietary intakes of infants from 3 to 12 months of age. AB - Data from a study of mother-infant pairs of low socioeconomic status living in Albany County, NY, were analyzed to determine the influence of diet and nutrition on the blood lead level of infants during the first year of life. Children's diets were assessed at 3-month intervals using a 24-h diet recall as reported by the primary caregiver. The potential impact of dietary consumption of protein, iron, zinc, calcium, vitamin D, and fat, as well as serum vitamin D and ferritin on blood lead levels at 6 and 12 months of age was examined with multivariable statistical analyses, controlling for other influences on lead levels. Neonates' blood lead levels were low at birth (geometric mean=1.6 microg/dL), and none were elevated (> or = 10 microg/dL). By 12 months, the mean blood lead for this sample was 5.1 microg/dL, and 18% of the sample had an elevated blood lead level. We observed significant inverse relationships between infants' 6-month lead level and their intake of zinc, iron, and calcium. At 12 months, low iron intake continued to be associated with higher lead levels, although zinc and calcium did not. Protein had a paradoxical effect, being associated with lower lead at 6 months, but higher lead at 12 months. Serum vitamin D and ferritin were not associated with lead levels, nor was vitamin supplement use. The results reported here emphasize the value of key minerals in the diet to reduce lead absorption during early infancy. PMID- 15364594 TI - Transition of cord blood lead level, 1985-2002, in the Taipei area and its determinants after the cease of leaded gasoline use. AB - Lead has long been of concern for its toxicity, impairment of neurobehavioral and cognitive development, and electrophysiological deficits in children, even at levels less than 10 microg/dL. The present study was conducted to elucidate the extent of cord blood lead level decline in the Taipei area from 1985 to 2002 and to explore the factors affecting the cord blood lead level after the cease of leaded gasoline use. In the current study period of 2001-2002, 184 of 1310 newborns delivered in the Taipei Municipal Women and Children Hospital between September 2001 and August 2002 were eligible and randomly selected to participate in this study. Neither of their parents had an occupational lead exposure history. At each delivery, a sample of 5-10 mL umbilical cord blood was collected for lead determination by graphite furnace atomic absorption spectrometry. The cord blood lead level of the newborns in the current study period averaged 2.35 +/- 1.12 microg/dL. Together with the cord blood lead averages of 7.48 +/- 2.25 and 3.28 +/- 1.52 microg/dL obtained from two previous surveys conducted in 1985 1987 and 1990-1992, respectively, the cord blood lead level was significantly decreased (P < 0.005). It is estimated that such a reduction in cord blood lead from 7.48 to 2.35 microg/dL for each year's cohort of 260,000 newborns in Taiwan might benefit the economics, ranging from US$8.9 billion to US$12.1 billion by improving the worker productivity. For the time period from 1985 to 2002, there were consistent transition patterns among the yearly fluctuations of air lead level, leaded gasoline consumption, lead content in gasoline, estimated lead amount emitted from the consumed leaded gasoline, and average cord blood lead levels of the three respective study periods. Additionally, every 0.1-g/L reduction in lead content in gasoline might lead to a lowering of cord blood lead level by 1.78 microg/dL. Furthermore, at low level of around 2 microg/dL, a multiple regression analysis demonstrated that economic status was the most influential factor for cord blood lead variation (P = 0.0061) while the maternal working month during her pregnancy was retained in the model with borderline effect (P = 0.0625). After accounting for the effect of leaded gasoline on the cord blood lead level, future study to differentiate the primary contributors for the low-level cord blood lead variation around 1-2 microg/dL is warranted. PMID- 15364595 TI - Multipathway organophosphorus pesticide exposures of preschool children living in agricultural and nonagricultural communities. AB - Environmental measurements of organophosphorus (OP) pesticides were conducted in the homes of 13 children, who lived either in the Seattle metropolitan area or in the agricultural region of Washington State, to ascertain exposure through multiple pathways. Each home was sampled for two 24-h periods during two seasons, summer and fall. Samples included 24-h indoor air, drinking water, soil, house dust, and hand and toy wipes and 24-h duplicate diets. At least one OP pesticide was found in each of the matrices sampled except for drinking water. Half of the indoor air samples contained detectable levels of chlorpyrifos or diazinon. Detectable levels of diazinon and azinphosmethyl in house dust were found in most of the agricultural homes, whereas only diazinon was found in the Seattle homes in the summer. Quantifiable chlorpyrifos and azinphosmethyl were found on either agricultural children's hands or their toys. These findings suggest different exposure pathways for children living in agricultural and nonagricultural regions. PMID- 15364596 TI - Lung oxidative response after acute coal dust exposure. AB - Coal dust exposure can induce an acute alveolar and interstitial inflammation that can lead to chronic pulmonary diseases. The objective of this study was to describe the acute and later effects of acute coal dust exposure in lung parenchyma and the involvement of reactive oxygen species in coal dust effects. Forty-eight male Wistar rats (200-250 mg) were separated into four groups: 48 h, 7 days, 30 days, and 60 days after coal dust instillation. Gross mineral coal dust (3 mg/0.5 mL saline) was administered directly in the lungs of the treatment group by intratracheal instillation. Control animals received only saline solution (0.5 mL). Lipid peroxidation was determined by the quantity of thiobarbituric acid-reactive species (TBARS), oxidative damage to protein was obtained by the determination of carbonyl groups, the total radical-trapping antioxidant parameter (TRAP) was estimated by luminol chemoluminescence emission, catalase activity was measured by the rate of decrease in hydrogen peroxide, and superoxide dismutase activity was assayed by the inhibition of adrenaline autooxidation. Histological evaluation of coal dust-treated rats demonstrated an inflammatory infiltration after 48 h of the exposure. Initially, this was a cellular infiltration suggestive of lymphocyte infiltration with lymphoid hyperplasia that remained until 7 days after induction. This initial response was followed by a chronic inflammatory infiltration characterized by aggregates of macrophages 30 days after induction. This inflammatory response tended to resolve 60 days after induction, being similar to that of control animals. During both the acute and chronic phases of lung inflammation we observed a decrease in the TRAP in the lung of coal dust-exposed animals compared to that in control animals. We also observed an activation of superoxide dismutase 60 days after coal dust exposition. TBARS were increased 60 days after coal dust exposure and protein carbonyl groups increased at all times after coal dust exposure (48 h, 7 days, 30 days, and 60 days). These data suggested a biphasic inflammatory response and the involvement of oxidative damage in coal dust-induced lung damage. PMID- 15364597 TI - Oxidation damage of sulfur dioxide on testicles of mice. AB - The effects of sulfur dioxide (SO2) on levels of thiobarbituric acid reactive substances (TBARS), levels of reduced glutathione (GSH), and the activities of Cu,Zn-superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were investigated in testicles of Kunming albino male mice. SO2 at different concentrations (22, 56, and 112 mg/m3) was administered to animals of SO2 groups in different exposure chambers for 6 h/day for 7 days, while control groups were exposed to filtered air under the same conditions. Our results show that SO2 caused lipid peroxidation and changes in antioxidative status in testicles of mice. Exposure to SO2 at all concentrations tested significantly increased TBARS levels in testicles of mice. SO2 at all concentrations tested tended to decrease activities of SOD and GPx enzymes and levels of GSH relative to control animals, but only the decreases in SOD and GPx activities caused by SO2 exposures of higher concentrations were statistically significant. SO2 at all concentrations tested tended to increase activities of CAT relative to control animals, but the increases of CAT activities caused by SO2 exposures of low concentrations (22 and 56 mg/m3) were statistically significant. These results lead to the conclusion that SO2 exposure can cause oxidative damage to testicles of male mice, and SO2 is a toxin to the reproductive system of mammals, not only to the respiratory system. Further work is required to understand the toxicological role of SO2 in reproduction organs or even sperm from humans and animals. PMID- 15364598 TI - Antiproliferative effect of butyltin in MCF-7 cells. AB - Humans are exposed to tributyltin compounds primarily through the intake of marine food. Previous reports on toxic effects to humans are limited to a few in vitro studies giving conflicting results regarding their effects on the aromatase enzyme and androgen receptor (AR) responses. The present study evaluates the estrogenic potential of three butyltin compounds (mono-, di-, and tributyltin) in an in vitro system based on the E-Screen assay. None of the butyltin compounds tested was estrogenic in the concentration range assayed (0.01-1000 nM). However, both dibutyltin dichloride (DBT) (500 nM) and tributyltin chloride (TBT) (10 nM) inhibited 17beta-estradiol-induced cell proliferation. DBT (500 nM) and TBT (10 nM) also significantly reduced testosterone-induced cell proliferation, and the inhibition by TBT was rescued by increasing the concentration of testosterone. The present study did not confirm the inhibition of aromatase as the mechanism for an endocrine effect of butyltin compounds; moreover, the inhibition of cell proliferation by DBT and TBT occurred at concentrations at which no cytotoxicity was observed. The exact mechanism by which TBT and DBT inhibit cell proliferation remains unexplained, but it might be essentially independent of the estrogen receptor. Therefore, these compounds may not be termed classical endocrine disruptors, but rather as compounds that cause a functional anti-estrogenic response. PMID- 15364599 TI - Marginal nutritional status of zinc, iron, and calcium increases cadmium retention in the duodenum and other organs of rats fed rice-based diets. AB - Dietary minerals Zn, Fe, and Ca are antagonistic to Cd absorption. We showed earlier that rats fed a rice-based diet with a marginal content of these nutrients absorbed more Cd than rats fed adequate Zn-Fe-Ca (Environ. Sci. Technol., 36 (2002) 2684-2692). The present experiment was designed to determine the effects of marginal dietary Zn, Fe, and Ca on the uptake and turnover of Cd in the gastrointestinal tract. Two groups of weanling female rats (six per treatment) were fed a diet containing 40% cooked, dried rice containing 0.6 mg Cd/kg. The diet of one group contained adequate Zn (35 mg/kg), Fe (30 mg/kg), and Ca (5000 mg/kg), while that of the other group contained marginal Zn (6 mg/kg), Fe (9 mg/kg), and Ca (2500 mg/kg). Rats were fed for 5 weeks and then orally dosed with 1g of diet containing rice extrinsically labeled with 109Cd. From 0.25 to 64 days after dosing, 109Cd and total Cd concentrations were determined in intestinal segments. Shortly after dosing, 109Cd, as a percentage of the dose, was about 4 times higher in the duodenum of marginally fed rats than in that of control rats (10% vs 40%, respectively). Sixty-four days after dosing, 109Cd was 10 times higher in marginally fed rats than in controls; however, of the amount at day 1, <0.1% remained at day 64. After 5 weeks, the concentration of elemental Cd in the duodenum of the marginally fed rats was 8 times higher than that of control rats (24 microg/g dry wt. vs 2.9 microg/g dry wt., respectively). Cd concentrations in liver and kidney were 5 times higher in the marginally fed rats than those in controls (liver, 0.81 microg/g dry wt. vs 0.14 microg/g dry wt.; kidney, 4.7 microg/g dry wt. vs 0.92 microg/g dry wt., respectively). These data suggest that marginal intakes of Zn, Fe, and Ca cause the accumulation of Cd in the duodenum, which results in a greater rate of Cd absorption and a greater accumulation in the internal organs. Results are discussed in relation to mineral nutrient status and risk assessment of Cd in natural food sources. PMID- 15364600 TI - Toxicity of azodrin on the morphology and acetylcholinesterase activity of the earthworm Eisenia foetida. AB - The acute toxicity of azodrin (monocrotophos, an organophosphorus insecticide) was determined on a soil organism, Eisenia foetida. The median lethal concentrations (LC50) were derived from a 48-h paper contact test and from artificial soil tests. The LC50 of azodrin in the paper contact test was 0.46 +/- 0.1 microg cm(-2) (23 +/- 6 mg L(-1)) and those in the 7- and 14-day artificial soil tests were 171 +/- 21 and 132 +/- 20 mg kg(-1), respectively. The neurotoxic potentiality of azodrin was assessed by using a marker enzyme, acetylcholinesterase (AChE; EC 3.1.1.7) in both in vitro and in vivo experiments. The progressive signs of morphological destruction are correlated with percentage inhibition of AChE in the in vivo experiments. The kinetics of AChE activity in the presence and absence of azodrin indicated that the toxicant is competitive in nature. This study demonstrated that azodrin causes concentration-dependent changes in the morphology and AChE activity of the earthworm E. foetida. PMID- 15364601 TI - Interactions between particulate air pollution and temperature in air pollution mortality time series studies. AB - In many community time series studies on the effect of particulate air pollution on mortality, particulate air pollution is modeled additively. In this study, we investigated the interaction between daily particulate air pollution and daily mean temperature in Cook County, Illinois and Allegheny County, Pennsylvania, using data for the period 1987-1994. This was done through the use of joint particulate air pollution-temperature response surfaces and by stratifying the effect of particulate air pollution on mortality by temperature. Evidence that the effect of particulate air pollution on mortality may depend on temperature is found. However, the results were sensitive to the number of degrees of freedom used in the confounder adjustments, the particulate air pollution exposure measure, and how the effects of temperature on mortality are modeled. The results were less sensitive to the estimation method used--generalized linear models and natural cubic splines or generalized additive models and smoothing splines. The results of this study suggest that in community particulate air pollution mortality time series studies the possibility of an interaction between daily particulate air pollution and daily mean temperature should be considered. PMID- 15364602 TI - Impact of cadmium exposure on male sex hormones: a population-based study in China. AB - The objective of this study was to investigate the possible effects of environmental cadmium (Cd) exposure on the levels of serum sex hormones in a Chinese population group. A total of 263 male volunteers were included. Blood samples were collected for the determination of serum testosterone (T), measured by radioimmunoassay, and follicle stimulating hormone (FSH) and luteinizing hormone (LH), both measured by enzyme immunoassays. Urinary and blood Cd were analyzed by atomic absorption spectroscopy (AAS). We found a dose-response relationship between urinary Cd excretion and the prevalence of abnormally high serum T levels, but, through multiple regression analysis, we could not trace exposure to Cd as a significant determinant of serum T levels. Exposure to Cd also failed to influence the levels of FSH and LH in serum. In contrast, we found that age, body mass index (BMI), and smoking habits are significant determinants of FSH and LH and of T and LH, respectively. We conclude that oral Cd exposure is not a critical determinant of hormone homeostasis in males, but lifestyle and some biological factors, such as age and BMI, are important. The relationship found between urinary Cd and high T levels may be of importance for male reproductive morbidity and should be investigated further. PMID- 15364603 TI - Trihalomethanes and associated potential cancer risks in the water supply in Ankara, Turkey. AB - The occurrence of trihalomethanes (THMs) in the water supply in the Ankara, Turkey was investigated. Total THMs and total organic carbon measurements were carried seasonally in the samples collected form 22 different districts along with the samples taken from the Ivedik Water Treatment Plant serving 90% of the city. The average summer nonpurgeable organic carbon (NPOC) concentration in the raw water was 4.2 mg/L, and the NPOC removal achieved in the treatment plant was 31%. The concentration of total THMs ranged from 25 to 74 microg/L, from 28 to 73 microg/L, and from 25 to 110 microg/L in winter, spring, and summer, respectively. In all of the samples chloroform existed at the highest concentrations, while bromoform was almost absent. The total THM concentrations were highest in summer for all districts. However, none of the concentrations detected exceeded the USEPA's Stage I limit of 80 microg/L and the EU's limit of 100 microg/L. However, the total THM level in 64% of the districts exceeded the USEPA's Stage II limit of 40 microg/L. The risk estimations carried out indicated that each year 1 of the 5 million Ankara residents could get cancer from the daily intake of water, mainly because of exposure to chloroform through oral ingestion. PMID- 15364604 TI - Osteoporosis and renal dysfunction in a general population exposed to cadmium in China. AB - Osteoporosis is a common metabolic disease characterized by low bone mass and microarchitectural deterioration of bone tissue. Many factors are involved in the occurrence of osteoporosis. Cadmium can cause both osteomalacia and osteoporosis and these effects have long been investigated through various epidemiological or experimental studies. The present study examines a possible relationship between cadmium nephropathy and its effects on the skeleton in populations living in a polluted area in southeast China. Monophoton absorptiometry was used to measure bone mineral density in the population and the Z score (the number of SD from the difference between the measured bone density of the individual and the group mean value for sex- and age-matched controls) was introduced to define osteoporosis (Z score < -2). Osteoporosis caused by cadmium exposure was demonstrated in this study on a general population environmentally exposed to cadmium in China. It was found that there were significant differences in the prevalence of osteoporosis among the different urinary cadmium groups (chi2 = 18.84, P = 0.0008). The linear trend test gave chi2 = 16.281, P = 0.00005. There was a dose-response relationship between cadmium exposure (urinary cadmium) and prevalence of osteoporosis. Of 31 subjects with osteoporosis, 23 subjects were suffering from renal dysfunction. The prevalence of renal dysfunction (74.19%) was significantly higher than that in those without osteoporosis (chi2 = 16.53, P < 0.001). Stratum analysis was performed to further assess the relationship between bone damage and renal impairment caused by cadmium. There was a significant difference between those with and without tubular damage (chi2 = 19.92, P = 0.000) but not in those with and without glomerular damage (chi2 = 0.08, P = 0.114). This showed that glomerular dysfunction plays a smaller role than tubular dysfunction in the causation of bone damage. It was found that the prevalence of osteoporosis increases with increasing values of parameters of tubular damage. Osteoporosis caused by cadmium is thus related to kidney dysfunction and especially to tubular damage and its severity but not to glomerular damage. The present study has thus demonstrated the combined adverse effects (osteoporosis and renal dysfunction) caused by environmental exposure to cadmium for the first time in Asia outside the endemic area in Japan. PMID- 15364605 TI - Molecular basis of AV block and cardiac malformations. PMID- 15364606 TI - Mutant cardiac ryanodine receptors and ventricular arrhythmias: is 'gain-of function' obligatory? PMID- 15364607 TI - Cytokines and late preconditioning. PMID- 15364608 TI - Heart failure with preserved ejection fraction. Diastolic dysfunction, subtle systolic dysfunction, systolic-ventricular and arterial stiffening, or misdiagnosis? PMID- 15364609 TI - Genes, stem cells and biological pacemakers. AB - The advent of gene therapy and cell therapy has led to reconsideration of standard therapies for cardiac disease. One such area of reconsideration is that of the cardiac pacemaker, which has been the mainstay of treatment for high degree heart block and sinoatrial node dysfunction. Over the past five years, gene therapy has been used to explore the overexpression of beta(2)-adrenergic receptors, the down-regulation of inward rectifier current, and the overexpression of pacemaker current as potential sources of biological pacemakers. Cell therapy approaches have explored the "forcing" of embryonic stem cells to evolve along cardiac (and specifically pacemaker) cell lines and the use of adult mesenchymal stem cells as platforms for delivery of specific gene therapies. This review considers the strengths and weaknesses of each of the approaches used to date and attempts to look to the future of biological alternatives to electronic pacemakers. PMID- 15364610 TI - Matricellular proteins in the heart: possible role during stress and remodeling. AB - Matricellular proteins are extracellular matrix proteins that modulate cell matrix interactions and cell function, and do not seem to have a direct structural role. The family includes tenascin-C (TN-C), tenascin-X (TN-X), osteonectin, osteopontin, thrombospondin-1 (TSP1) and thrombospondin-2 (TSP2). Expression of matricellular proteins is high during embryogenesis, but almost absent during normal postnatal life. Interestingly, it re-appears in response to injury. Left ventricular remodeling is a complicated process that occurs in the stressed heart, and is still not completely understood. Several members of the matricellular protein family, like tenascin-C, osteopontin, and osteonectin are up-regulated after cardiac injury. Therefore, this group of proteins may have crucial functions in the heart coping with stress. This review will focus on the expression, regulation and function of these matricellular proteins, and will discuss the crucial functions that these proteins might exert during remodeling of the stressed heart. PMID- 15364611 TI - Pharmacogenomics of heart failure -- focus on drug disposition and action. AB - Heart failure represents the composite endpoint of various cardiovascular disorders. Advanced pharmacotherapy resulted in significant improvement of overall survival, however with highly variable outcome, possibly due to genetic modification of drug disposition and action. This review highlights the role of genetic polymorphisms in systems responsible for disposition of drugs, used in heart failure patients (e.g. the polymorphic drug metabolizing enzymes such as cytochrome P450 enzymes, as well as polymorphic ATP-membrane transporters like P glycoprotein (P-gp)). In addition, genetic variants in physiological systems, being target of drug action, particularly beta-adrenergic receptors, the renin angiotensin-aldosterone system (RAAS)- and endothelin system, and the endothelial nitrogene monoxide (NO) synthase are reviewed. The current situation in pharmacogenomics of heart failure with respect to drug disposition and action is characterized by multiple studies investigating single components of a complex system. Therefore, overall conclusions regarding treatment and/or outcome of heart failure patients based on individual genetic traits require large prospective trials allowing for simultaneous assessment of multiple genetic variants in different systems. Using advanced screening technologies, such trials can be carried out in the near future. PMID- 15364612 TI - Biochemical analyses of eight NKX2.5 homeodomain missense mutations causing atrioventricular block and cardiac anomalies. AB - OBJECTIVE: There has been considerable interest in understanding determinants of the diverse cardiac phenotypes associated with heterozygous NKX2.5 mutations. We hypothesized that analysis of functional properties of NKX2.5 mutant proteins would result in the ability to classify mutations according to function in a scheme that would help to clarify genotype-phenotype correlations. We analyzed missense mutations in the conserved homeodomain. METHODS: We studied in vitro biochemical characteristics, including nuclear localization, DNA binding, transcriptional activation and protein-protein interaction with transcriptional partners (GATA4, TBX5 and NKX2.5 itself), of eight homeodomain missense mutations. Associated phenotypes include atrioventricular (AV) block (98% penetrance), atrial septal defect (83% penetrance), and additional varied heart malformations. RESULTS: Mutations were present at varied homeodomain locations in the putative nuclear localizing signal (1), helix 2 (1), a turn between helix 2 and 3 (1) and helix 3 (5); a spectrum of biochemical phenotypes was observed. All mutants localized to the nuclei but some exhibited anomalous nuclear distribution. While all mutants exhibited markedly decreased DNA binding and reduced transcriptional activation, interaction with transcriptional partners was varied. CONCLUSION: Each mutant protein had a unique spectrum of observed properties, but our data show that while dominant negative properties could be demonstrated in vitro, the best correlation with clinical phenotypes resulted from the markedly reduced DNA binding shared by all eight homeodomain mutations. This suggests that the principle determinant of the two most common phenotypes associated with homeodomain missense mutations is the total dose of NKX2.5 capable of binding to DNA. PMID- 15364613 TI - Functional heterogeneity of ryanodine receptor mutations associated with sudden cardiac death. AB - OBJECTIVES: Point mutations in the cardiac ryanodine receptor (RyR2) mediate abnormal intracellular Ca(2+) release and are associated with stress-induced ventricular tachycardia (VT), leading to sudden cardiac death (SCD). Although the precise molecular basis of RyR2 dysfunction in SCD remains controversial, there is consensus that the mutations characterised to date all exhibit gain-of function Ca(2+) release properties following cell stimulation. We investigated the functional impact of a distinct set of SCD-linked RyR2 mutations (L(433)P, N(2386)I, R(176)Q/T(2504)M) on intracellular Ca(2+) handling. METHODS: We expressed full-length recombinant human wild-type (WT) and SCD-linked RyR2 mutations in human embryonic kidney (HEK) cells, and profiled the spatial and amplitude characteristics of caffeine-evoked Ca(2+) release through homo tetrameric channels in living cells using rapid confocal laser scanning microscopy. RESULTS: Analysis of the precise mode of Ca(2+) release in HEK cells expressing RyR2 mutants demonstrated profound differences when compared with WT channels. The SCD-linked RyR2 mutations characterised in this study exhibited heterogeneous Ca(2+) release profiles, including the novel observation that one of the mutants, (L(433)P), exhibited a marked reduction in sensitivity to channel activation. However, all SCD-linked RyR2 mutations characterised in this study resulted in an increased duration of elevated cytoplasmic Ca(2+) levels following channel activation. CONCLUSIONS: Our live cell-based data demonstrates functional heterogeneity of Ca(2+) release through SCD-linked RyR2 mutants, suggesting that the mechanistic basis of RyR2 dysfunction in SCD may be more complex than previously anticipated. These findings may have profound consequences for the therapeutic modulation of RyR2 in stress-induced VT and SCD. PMID- 15364615 TI - Mechanisms of heart failure with well preserved ejection fraction in dogs following limited coronary microembolization. AB - OBJECTIVE: It has been suggested that in some settings, heart failure (HF) may occur with normal ejection fraction (EF) as a consequence of undetected systolic dysfunction. However, others have argued that this can only occur in the presence of diastolic dysfunction. We therefore sought to determine the contribution of diastolic dysfunction in an animal model of HF with normal EF. METHODS AND RESULTS: Limited myocardial injury was induced in 21 dogs chronically instrumented to measure hemodynamics and LV properties by daily coronary microembolization ( approximately 115 microm beads) until LV end diastolic pressure (LVEDP) was > or =16 mm Hg. Nine dogs developed HF within 16+/-6 days (LVEDP 12+/-2 vs. 21+/-2 mm Hg, p<0.001) with no significant change in dP/dt(max) (2999+/-97 vs. 2846+/-189 mm Hg/s), mean arterial pressure (103+/-4 vs. 100+/-4 mm Hg), EF (57+/-5% vs. 53+/-4%) or E(es) (end-systolic elastance, 3.1+/-0.9 vs. 2.9+/-0.8 mm Hg/ml) but with an approximately 10 ml increase in V(o) (14+/-12 vs. 25+/-16 ml; p<0.01). The EDPVR and time constant of relaxation (tau, 25+/-3 vs. 28+/-3 ms) did not change significantly. These animals were hemodynamically stable out to 3 1/2 months. Neurohormonal activation occurred (elevations of NE, AngII, BNP) and there was intravascular volume expansion by approximately 16% (p<0.05). CONCLUSIONS: A small amount of myocardial injury can lead to neurohormonal activation with intravascular volume expansion and elevation of LVEDP in the absence of reductions in dP/dt(max) or EF and without diastolic dysfunction. Thus, HF with preserved EF does not a priori equate with diastolic heart failure. PMID- 15364614 TI - IL-6 plays an obligatory role in late preconditioning via JAK-STAT signaling and upregulation of iNOS and COX-2. AB - OBJECTIVE: We sought to determine whether interleukin (IL)-6 modulates myocardial infarction or the late phase of preconditioning (PC). METHODS: Wild-type and IL 6(-/-) mice underwent a 30-min coronary occlusion followed by 24 h of reperfusion with or without six cycles of coronary occlusion/reperfusion 24 h earlier. Myocardial IL-6 protein expression, activation of Janus kinase (JAK) 1 and JAK2, and signal transducers and activators of transcription (STAT) 1 and STAT3 after ischemic PC protocol were examined. The expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 was determined 24 h after the PC ischemia. RESULTS: In preconditioned wild-type mice, infarct size was reduced from 60.5+/-2.6% of the risk region to 33.5+/-3.6%, indicating a late PC effect. In nonpreconditioned IL-6(-/-) mice, infarct size was similar to that observed in wild-type mice (59.9+/-3.8%), indicating that the deletion of IL-6 has no effect on infarct size. However, in preconditioned IL-6(-/-) mice, infarct size was not reduced (65.1+/-3.1%), indicating that the infarct-sparing effect was completely abrogated. Ischemic PC increased the expression of IL-6 in the cytoplasm of cardiomyocytes in the ischemic/reperfused zone. In IL-6(-/-) mice, the ischemic PC-induced activation of JAK1 and JAK2 and STAT1 and STAT3 was significantly reduced, and the increase in iNOS and COX-2 protein expression 24 h after the PC ischemia was markedly attenuated. CONCLUSION: IL-6 does not modulate myocardial infarct size in naive myocardium. However, following a PC stimulus, IL-6 is obligatorily required for the activation of the JAK-STAT pathway, the ensuing upregulation of iNOS and COX-2 (co-mediators of late PC), and the development of a cardioprotective phenotype. PMID- 15364616 TI - Postnatal development of atrial repolarization in the mouse. AB - OBJECTIVES AND METHODS: This study examines postnatal development of action potential duration (APD) and voltage-dependent K(+) currents in mouse atrial myocytes and compares the expression levels of corresponding K(+) channels between adult and neonatal mouse atrial tissues. APD and K(+) currents were compared between atrial myocytes isolated from postnatal Day-1, Day-7, Day-20, and adult mice. RESULTS: All K(+) currents examined underwent significant up regulation during postnatal life in mouse atrium, resulting in a dramatic shortening of the APD. The ultrarapid delayed rectifier (I(Kur)) was absent in the developing mouse heart and only contributed to repolarization in the adult mouse atrium, whereas the density of the other K(+) currents increased earlier during the developmental period. Indeed, the major changes in the expression of the inward rectifier current (I(K1)) occurred within the first week of life, the density of the Ca(2+)-independent transient outward K(+) current (I(to)) gradually increased while the development of the steady-state outward K(+) current (I(ss)) was completed within the first 3 weeks of life. Results of RNase protection assay and Western blot analysis confirmed that the postnatal development of the mouse atrial K(+) currents correlates with an increase in expression levels of underlying K(+) channel isoforms. CONCLUSION: These findings indicate that in mouse atrium, each K(+) current exhibits a specific postnatal development, suggesting that regulatory factors taking place during development are major determinants of the functional role of K(+) channels in cardiac repolarization. The mouse atrium is, therefore, a very interesting model to gain information on the mechanisms regulating K(+) channel activity. PMID- 15364617 TI - Regulation of glucose transporter expression in cardiac myocytes: p38 MAPK is a strong inducer of GLUT4. AB - OBJECTIVE: In vivo differentiation of cardiac myocytes is associated with downregulation of the glucose transporter isoform GLUT1 and upregulation of the isoform GLUT4. Adult rat cardiomyocytes in primary culture undergo spontaneous dedifferentiation, followed by spreading and partial redifferentiation, which can be influenced by growth factors. We used this model to study the signaling mechanisms modifying the expression of GLUT4 in cardiac myocytes. RESULTS: Adult rat cardiomyocytes in primary culture exhibited spontaneous upregulation of GLUT1 and downregulation of GLUT4, suggesting resumption of a fetal program of GLUT gene expression. Treatment with IGF-1 and, to a minor extent, FGF-2 resulted in restored expression of GLUT4 protein and mRNA. Activation of p38 MAPK mediated the increased expression of GLUT4 in response to IGF-1. Transient transfection experiments in neonatal cardiac myocytes confirmed that p38 MAPK could activate the glut4 promoter. Electrophoretic mobility shift assay in adult rat cardiomyocytes and transient transfection experiments in neonatal cardiac myocytes indicated that MEF2 was the main transcription factor transducing the effect of p38 MAPK activation on the glut4 promoter. CONCLUSION: Spontaneous dedifferentiation of adult rat cardiomyocytes in vitro is associated with downregulation of GLUT4, which can be reversed by treatment with IGF-1. The effect of IGF-1 is mediated by the p38 MAPK/MEF2 axis, which is a strong inducer of GLUT4 expression. PMID- 15364618 TI - Ischemic preconditioning attenuates calpain-mediated degradation of structural proteins through a protein kinase A-dependent mechanism. AB - OBJECTIVES: It has been shown that sarcolemmal rupture can occur during reenergization in cardiomyocytes in which previous ischemia has induced sarcolemmal fragility by calpain-dependent hydrolysis of structural proteins. We tested the hypothesis that attenuated calpain activation contributes to the protection against reperfusion-induced cell death afforded by ischemic preconditioning (IPC), and investigated the involvement of protein kinase A (PKA) in this effect. METHODS: Calpain activity and degradation of different structural proteins were studied along with the extent of necrosis in isolated rat hearts submitted to 60 min of ischemia and 30 min of reperfusion with or without previous IPC (two cycles of 5 min ischemia-5 min reperfusion), and the ability of different treatments to mimic or blunt the effects of IPC were analyzed. RESULTS: IPC accelerated ATP depletion and rigor onset during ischemia but reduced LDH release during reperfusion by 69% (P<0.001). At the end off reperfusion, calpain activity was reduced by 66% (P<0.001) in IPC, and calpain-dependent degradation of sarcolemmal proteins was attenuated. Addition of the calpain inhibitor MDL 28170 mimicked the effects of IPC on protein degradation and reduced LDH release by 48% (P<0.001). The effects of IPC on calpain, alpha-fodrin, and LDH release were blunted by the application of the PKA inhibitor H89 or alprenolol during IPC, while transient stimulation of PKA with CPT-cAMP or isoproterenol before ischemia attenuated calpain activation, alpha-fodrin degradation, and markedly reduced LDH release (P<0.001). In hearts exposed to Na(+)-free perfusion, IPC attenuated calpain activation by 67% (P<0.001) and reduced by 56% (P<0.001) LDH release associated to massive edema occurring during Na(+) readmission without modifying its magnitude. CONCLUSION: These results are consistent with PKA dependent attenuation of calpain-mediated degradation of structural proteins being an end-effector mechanism of the protection afforded by IPC. PMID- 15364619 TI - Reciprocal regulation of angiopoietin-1 and angiopoietin-2 following myocardial infarction in the rat. AB - OBJECTIVE: This study sought to characterize changes in the angiopoietin system in a rat model of myocardial infarction (MI). BACKGROUND: Angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) bind to the endothelial-specific receptor tyrosine kinase, TIE-2. Ang-2 has been suggested to be an antagonist of TIE-2, possibly acting to release endothelial cells from the tonic stabilizing influence of Ang 1. However, on prolonged exposure, Ang-2 has been shown to acquire agonistic activity at TIE-2, raising the possibility that this isoform may play a direct role in neovascularization. METHODS: Sprague-Dawley rats were subjected to left coronary ligation and myocardial tissues were harvested from the infarct and peri infarct regions, or from non-infarcted myocardium. Changes in gene expression were determined by RT-PCR and confirmed by Northern analysis. Changes in protein expression were confirmed by Western analysis and immunocytochemistry, and TIE-2 activity was determined by immunoprecipitation with anti-TIE-2 and antiphosphotyrosine immunoblotting. RESULTS: At 24 h, Ang-1 mRNA and protein expression within the infarct and peri-infarct regions were decreased compared to non-infarcted myocardium, whereas Ang-2 mRNA levels were markedly increased and TIE-2 expression was unchanged. Immunohistochemical staining revealed Ang-1 and TIE-2 immunoreactivity localized to vascular endothelium. In the infarct territory, Ang-2 immunostaining was localized primarily to invading leukocytes at 24 h. At 1 week, Ang-1 expression was partially restored, whereas Ang-2 expression remained elevated. At the time of peak elevation in Ang-2, Tie2 phosphorylation was found to be markedly increased, consistent with receptor activation. CONCLUSIONS: Thus, myocardial ischemia induced by left coronary artery ligation resulted in a sustained increase in Ang-2 expression and a reciprocal decrease in Ang-1, consistent with a predominant role for Ang-2 in the angiogenic response to MI. PMID- 15364620 TI - Extraneuronal enzymatic degradation of myocardial interstitial norepinephrine in the ischemic region. AB - OBJECTIVE: Catechol O-methyltransferase (COMT) is believed to exert degradative action at high norepinephrine (NE) levels. Although COMT exists in cardiac tissues, the contribution of cardiac COMT activity to regional NE kinetics, particularly in ischemia-induced NE accumulation, remains unclear. We investigated the role of cardiac COMT in NE kinetics in the ischemic region. METHODS: We implanted a microdialysis probe into the left ventricular myocardium of anesthetized rabbits and induced myocardial ischemia by 60-min coronary artery occlusion. We monitored myocardial interstitial levels of NE and its metabolites in the presence and absence of a COMT inhibitor. We intraperitoneally administered entacapone (10 mg/kg) 120 min before control sampling. RESULTS: In control, entacapone increased interstitial dihydroxyphenylglycol (DHPG, intraneuronal NE metabolite by monoamine oxidase (MAO)) levels and decreased interstitial normetanephrine (NMN, extraneuronal NE metabolite by COMT) and 3 methoxy-4-hydroxyphenylglycol (MHPG, extraneuronal DHPG metabolite by COMT) levels, but did not change interstitial NE levels. Coronary occlusion increased NE levels to 165+/-48 nM at 45-60 min of occlusion. This increase was accompanied by increases in DHPG and NMN levels (11.3+/-1.1 and 9.3+/-1.3 nM at 45-60 min of occlusion). Entacapone augmented the ischemia-induced NE and DHPG responses (333+/-51 and 22.9+/-2.4 nM at 45-60 min of occlusion). In contrast, the ischemia induced NMN response was suppressed by entacapone (2.0+/-0.4 nM at 45-60 min of occlusion). Reperfusion decreased interstitial NE levels and increased interstitial DHPG and NMN levels. Entacapone suppressed changes in NE and NMN levels, but augmented the increase in dialysate DHPG. CONCLUSION: Myocardial ischemia evoked increases in myocardial interstitial NE and NMN levels. COMT inhibition augmented the increase in NE (substrate of COMT) levels and suppressed the increase in NMN (metabolite by COMT) levels. In the ischemic heart, COMT contributes to the removal of accumulated NE in the myocardium. PMID- 15364621 TI - T-cadherin upregulation correlates with cell-cycle progression and promotes proliferation of vascular cells. AB - OBJECTIVE: In vascular tissue, T-cadherin (T-cad) levels correlate with the progression of atherosclerosis, restenosis and tumour neovascularization. This study investigates whether T-cad influences proliferation of vascular cells. METHODS AND RESULTS: Cultures of human umbilical vein endothelial cells (HUVEC) and rat and human aortic smooth muscle cells (rSMC, hSMC) were used. T-cad was overexpressed in HUVEC and hSMC using an adenoviral expression system. In cultures released from G(1)/G(0) synchrony parallel immunoblot analysis of T-cad and cell cycle phase specific markers (p27(Kip1), cyclin D1, E2F1, PCNA, cyclin B) showed increased T-cad protein levels subsequent to entry into early S-phase with sustained elevation through S-and M-phases. T-cad was increased in G(2)/M phase (colchicine) synchronized cultures. In FACS-sorted cell populations, expression of T-cad in S-and G(2)/M-phase was higher than G(1)/G(0)-phase. Compared with empty-and LacZ-vector infected controls, HUVEC and hSMC overexpressing T-cad exhibited increased proliferation as assessed in enumeration and DNA synthesis assays. Additionally, following release from G(1)/G(0) synchrony, HUVEC and hSMC overexpressing T-cad enter S-phase more rapidly. Flow cytometry after BrdU/propidium labelling confirmed increased cell cycle progression in T-cad overexpressing cells. CONCLUSION: In vascular cells, T-cad is dynamically regulated during the cell cycle and its expression functions in the promotion of proliferation. T-cad may facilitate progression of proliferative vascular disorders such as atherosclerosis, restenosis and tumour angiogenesis. PMID- 15364622 TI - 7-Ketocholesterol induces reversible cytochrome c release in smooth muscle cells in absence of mitochondrial swelling. AB - OBJECTIVE: 7-Ketocholesterol, a major oxysterol in oxidized low-density lipoproteins in advanced atherosclerotic plaques, induces vascular smooth muscle cell (SMC) death. We investigated whether cytochrome c release participated in SMC death induced by 7-ketocholesterol and whether the processes were reversible. METHODS: SMC cultures derived from the rabbit aorta were exposed to 25 microM 7 ketocholesterol. Cytochrome c and Bax were studied by means of immunofluorescence and immunoblotting, apoptosis by the TUNEL technique and mitochondrial structure by transmission electron microscopy. RESULTS: 7-Ketocholesterol induced rapid upregulation of the proapoptotic protein Bax and its translocation from cytosol into the mitochondria (4 h). This was followed by mitochondrial cytochrome c release (65% at 8 h) into the cytosol, which was almost complete at 16 h. The mitochondria became spherical and ultracondensed, without showing signs of lysis. They clustered around the nucleus and were wrapped by wide cisternae of the rough endoplasmic reticulum. Cytochrome c release was not blocked by the pan-caspase inhibitor zVAD-fmk, in contrast to DNA fragmentation and SMC loss. Interestingly, upon removal of 7-ketocholesterol after 16 h and re-exposure to serum for 24 h, the mitochondrial cytochrome c content, their transmembrane potential and TUNEL labelling normalised and SMC loss decreased. However, none of these cell death markers was rescued when the SMCs had been exposed to the oxysterol for 24 h. CONCLUSION: The results indicate that cytochrome c release during oxysterol induced SMC apoptosis is not caspase-dependent and occurs as a result of a reversible mitochondrial conformational change rather than swelling and rupture of the outer membrane. The reversibility of these events suggests that the apoptotic cascade could be arrested before a point of no return. PMID- 15364623 TI - Inflammatory signaling pathway containing TRAF6 contributes to neointimal formation via diverse mechanisms. AB - OBJECTIVE: The purpose of this study was to investigate the contribution of inflammatory signaling containing tumor necrosis factor receptor-associated factor 6 (TRAF6) to neointimal formation in a balloon injury model of rabbit carotid artery. METHODS: Male Japanese white rabbits fed a normal diet were used. We transferred the dominant negative (DN) form of TRAF6 to a rabbit carotid artery that was subjected to balloon injury by in vivo electroporation method, and then evaluated its effect on intimal lesion formation after balloon injury. RESULTS: An expression plasmid vector containing the TRAF6 DN sequence was successfully transferred to arterial wall cells, and its inhibitory effect on inflammatory signaling was confirmed by the marked suppression of nuclear factor kappaB (NFkappaB) activity after injury. Morphometric analyses revealed significant inhibition of intimal lesion formation at 7 days after injury. Cell replication and accumulation of macrophages in the media were significantly decreased, and apoptosis was enhanced on day 2. Cell migration to the intima was suppressed on day 4. Extracellular signal-regulated kinase1/2 (ERK1/2) activity at 2 h after injury was also down-regulated. Interestingly, intimal cell replication was significantly blocked when TRAF6 DN was transfected at 7 days after injury. CONCLUSION: TRAF6 plays important roles in cell replication and migration, besides promotion of inflammatory cell infiltration and suppression of apoptosis. PMID- 15364624 TI - Tyrphostin AGL-2043 eluting stent reduces neointima formation in porcine coronary arteries. AB - OBJECTIVE: Tyrphostin AGL-2043 is a potent tricyclic quinoxaline inhibitor of PDGF beta-receptor tyrosine kinase (PTK), Kit, and Flt3. We have shown previously that selective inhibition of PDGF beta-receptor PTK by tyrphostins markedly reduces SMC proliferation and migration in vitro, reduces neointima formation in balloon-injured porcine femoral arteries, and reduces neointimal stenosis in stented porcine coronary arteries when administered intramurally within biodegradable nanoparticles. The present study was designed to determine the effect of AGL-2043 delivered from a stent-based, biodegradable polymeric coating on neointima formation in the porcine coronary artery model. METHODS AND RESULTS: Stents coated with biodegradable, polylactic/glycolic acid (PLGA) polymer, with (n=13) or without (n=11) 180 mcg AGL-2043 were implanted into the proximal LAD of 24 Sinclair mini-pigs (34+/-4 kg) to achieve a 1.1:1 stent/artery diameter ratio. The delivery of drug from stent to tissue was confirmed by high-performance liquid chromatography. After 28 days, histomorphometric analysis showed that in stent stenosis in animals treated with AGL-2043 was reduced by 50% (51+/-21% versus 26+/-10%, p=0.001), the absolute neointimal area was reduced by 44% (2.38+/-1.04 versus 1.31+/-0.43 mm(2), p=0.004), and the absolute luminal area was increased by 57% (2.19+/-1.09 versus 3.39+/-0.59 mm(2), p=0.003). There were no significant differences between control and AGL-2043 in injury score (1.24+/ 0.11 vs. 1.15+/-0.12, p=0.07) or inflammation score (1.19+/-0.35 vs. 1.07+/-0.33, p=0.41). Moreover, the difference in % in-stent stenosis between control and treated animals remained highly significant even after normalizing the % stenosis to the degree of injury (p=0.0008) or to the inflammation score (p=0.001). Mortality for this study was zero. Tissue concentration in segments 1 cm proximal and distal to the stents, were negligible or zero at 1 h, 24 h, and 4 weeks after stent implantation. CONCLUSION: Stent-based delivery of tyrphostin AGL-2043 from a biodegradable polymeric coating reduces in-stent neointimal hyperplasia in porcine coronary arteries by 50% after 28 days and preserves lumen area. Long term studies should be the next step in testing applicability to the human interventional setting. PMID- 15364625 TI - Inflammation-induced endothelial dysfunction involves reduced nitric oxide bioavailability and increased oxidant stress. AB - OBJECTIVES: Our aim was to investigate mechanisms of inflammation-induced endothelial dysfunction in humans. METHODS: Endothelial function in twenty-one healthy human volunteers was measured using forearm venous plethysmography before and 8 h after administration of typhoid vaccination to generate an inflammatory response. Basal and stimulated endothelial nitric oxide (NO) bioavailability was assessed by measurement of the responses to intra-arterial N(G)-monomethyl-l arginine (l-NMMA) and bradykinin, respectively. The effects of supplementation with l-arginine or ascorbic acid were assessed to probe the effects of substrate deficiency and oxidative stress, respectively. Systemic effects were determined by measuring cytokine response, total anti-oxidant status (TAOS) and urinary protein excretion. RESULTS: Vaccination induced a cytokine response, a fall in total anti-oxidant status and increased urinary albumin excretion (UAE). There was a reduction in the response to bradykinin (BK, P<0.005) and l-NMMA (P<0.0001) with no effect on the response to glyceryl trinitrate (GTN) and norepinephrine (NE). Following vaccination blood flow response to BK (but not GTN) was partially returned to pre-vaccine levels by infusion of ascorbic acid (P=0.01). Supplementation with l-arginine had no effect. CONCLUSION: Inflammation causes widespread endothelial dysfunction, reduces vascular NO bioavailability and increases oxidative stress. These actions are partially reversible with local anti-oxidants. These findings suggest a role for reactive oxygen species in inflammation-induced endothelial dysfunction. PMID- 15364626 TI - Long-term effects of repeated pregnancies (multiparity) on blood pressure regulation. AB - OBJECTIVE: Pregnancy is associated with profound alterations in the cardiovascular system, the long-term effects of which are unknown. Human epidemiological studies suggest that multiparity (multiple pregnancies) increases the risk of cardiovascular disease. The mechanisms underlying these findings remain to be elucidated. The objective of this study was to determine the long term effects of parity on cardiovascular regulation. METHODS: Pressor responses to phenylephrine (PE) and acute stress were compared in conscious age-matched repeatedly breed (RB) and virgin rats. Vascular compliance and reactivity of isolated resistance-sized mesenteric arteries were studies using pressure and wire myograph. RESULTS: We found that both exogenous PE and acute stress elicited greater pressor responses in RB than in aged-matched virgins. Pressure and wire myography also revealed that small mesenteric arteries from RB rats were less compliant than those from virgins (RB: 0.24+/-0.04 microm mm Hg(-1), n=6 vs. virgins: 0.63+/-0.06 microm mm Hg(-1), n=6; p< or =0.05) and were more sensitive to PE (EC(50) RB: 1.58+/-0.08 x 10(-6) M, n=10 vs. virgins: 2.05+/-0.09 x 10(-6) M, n=14; p< or =0.05). Removal of the endothelium abolished the difference in sensitivity. More specifically, the augmented vascular response of RB was both nitric oxide (NO) and cyclooxygenase dependent. By contrast, there was no difference in methacholine-induced vasodilation of phenylephrine-preconstricted vessels. CONCLUSION: Our results suggest that repeated pregnancies induce long term alterations in cardiovascular regulation due to changes in vascular compliance and endothelium-dependent vasoconstriction. We propose that such changes might influence the risk for cardiovascular disease in multiparous women. PMID- 15364627 TI - The federal ban on ephedrine dietary supplements:an important event for pain practitioners and patients. PMID- 15364628 TI - Cancer pain intensity measurements in outpatients: preferences and comparison of pain scales among patients, caregivers, physicians and nurses in southern India. AB - Pain is frequently encountered in outpatient oncology practice and its management often is inadequate. Effective analgesia often could be provided for these patients through simple practices like pain intensity monitoring, documentation and treatment accordingly. A survey was carried out among cancer patients attending outpatient pain clinic to evaluate scale preferences, comparison of different scales as well as proxy reporting by caregiver, health professionals, for pain management. The aim of the study was to determine preferences in Indian population and to compare our study subjects' preferences to those previously reported in other cultures. A total of 99 patients participated in the study. The majority preferred a visual analogue scale (VAS). The visual analogue scale and verbal descriptive scale (VDS) were found to be equally reliable pain rating tools. Proxy reports were found to equally representative of patient's pain. VAS was the preferred pain assessment tool. In the absence of VAS scale, VDS also could be used. Proxy reports could be relied upon for management of pain. PMID- 15364629 TI - Topical lidocaine patch therapy for myofascial pain. AB - An open label study of topical lidocaine 5% patches was conducted for myofascial pain management based on the hypothesis that electrical dysfunction is a component of myofascial pain and therefore sodium channel blockade may be useful in managing myofascial pain. The efficacy of topical lidocaine patch therapy for myofascial pain impact of the therapy on associated quality of life were investigated in the one-month trial. Principal outcome measures were Brief Pain Inventory- Short Form for pain intensity and quality of life score changes. Twenty-seven patients with moderate-severe myofascial pain were enrolled. Eighteen had low back pain. Two patients reported complete pain relief and 3 reported a lot of relief. Mean improvements for average pain intensity (7, 14, and 28 days), general activity (7 and 28 days), mood and sleep (7, 14, and 28 days), walking (14 and 28 days), and ability to work, relationships, and enjoyment of life (28 days) were significant (P < 0.05). These results suggest lidocaine patches may be useful in the management of myofascial pain. PMID- 15364630 TI - Bupreorphine:a new pharmacotherapy for opioid addictions treatment. AB - The federal Drug Abuse Treatment Act 2000 (DATA) opened a window of opportunity for patients with the disease of addiction by providing increased access to options for treatment. Previously, only methadone maintenance, approved for use only through specially regulated clinics, was available to treat opioid addiction. DATA allows any physician choosing to take a short specialty training course and become certified to prescribe buprenorphine. Buprenorphine and buprenorphine/ naloxone (Subutex, Suboxone) can be prescribed by certified physicians in a traditional office setting to treat patients with opioid dependence. Clinical studies indicate buprenorphine maintenance is as effective as methadone maintenance in retaining patients in substance abuse treatment and reducing illicit opioid use. Sublingual buprenorphine is more effective than clonidine or clonidine/naltrexone in short-term opioid detoxification treatment. Buprenorphine provides an additional tool to treat opioid addiction and improve the quality of lives of these patients. More physicians are needed to treat patients with addiction. DATA facilitates this by removing existing barriers and increasing access to treatment. PMID- 15364631 TI - Myths about controlling pain. AB - Myths and misinformation about pain and its management often lead to poor therapy for patients in pain. Nine common myths about pain and its management are described and refuted. PMID- 15364632 TI - American society of health-system pharmacists' pain management network. AB - The American Society of Health-System Pharmacists' (ASHP) Pain Management Network is one of several informal networking groups of pharmacist practitioners who specialize in a clinical practice area. Pharmacy practitioners are showing an increased interest in the management of pain because of expanding clinical roles, the Joint Commission on Accreditation of Health Care Organizations' development of pain management standards, and involvement with performance improvement and quality initiatives. The Pain Management Network is one of the American Society of Health-system Pharmacists (ASHP) Section of Clinical Specialists and Scientists specialty practice networks. The Network serves as an informal venue for practitioners interested in pain management to gather and share information. Pain Management Network Assemblies are held at each ASHP national meeting. Practitioners communicate between meetings via the Section's e-mail list server. Pharmacists who participate in the Pain Management Network have a variety of roles in both acute and chronic pain management. Their roles may be consultative in nature or they may be involved in the actual medication management as part of a collaborative drug therapy management agreement with a physician. Pharmacists are involved with acute pain, cancer-related pain, and chronic nonmalignant pain management. Current issues involve education of pharmacy students and practicing pharmacists as well as the use of meperidine as an analgesic agent. Compounding of non-sterile drugs for intraspinal use is another current issue. Access to evidence-based pain management resources may be difficult to locate and access but is essential to good pain management. The ASHP Pain Management Network has proven to be a valuable way for participants to gather and exchange new information and share in their clinical practice experiences. PMID- 15364633 TI - Thinking the unthinkable:the clinician as perpetrator of elder abuse in patients in pain. AB - Some advocates for improved pain management have devised and had initial success utilizing a controversial new weapon-the elder abuse claim. In California, two recent cases have been brought under the state's elder abuse statutes against physicians and health care institutions. The first lead to a much publicized jury verdict against a physician, and the second was recently settled as to all defendants, with related disciplinary actions against a treating physician and a skilled nursing facility by their respective regulatory agencies. This commentary reviews the phenomenon of litigating cases of undertreated pain, analyzes the most recent cases, and considers the implications of invoking elder abuse statutes in such cases. PMID- 15364634 TI - Opioid insights:opioid-induced hyperalgesia and opioid rotation. AB - Opioid analgesics are an irreplaceable component of pharmacotherapy of numerous pain-producing conditions. Clinicians and patients must contend with the imperfect nature of this class of drugs, trying to balance benefits and burdens on a continual basis. New literature related to evidence-based selection of opioids and the neurobiological phenomenon of opioid induced hyperalgesia are reviewed. A matrix describing critical elements in the selection of opioid analgesics, both for initial therapy and for opioid rotation, is presented. PMID- 15364635 TI - Methods to measure patient compliance with medication regimens. AB - Successful pharmacotherapy requires patient compliance with the therapeutic regimen. The rate of non-compliance has been reported to be as low as 15% in some studies and the costs of non-compliance have been estimated to be between $396 and 792 million. Compliance, adherence, and persistence are defined. Subjective and objective methods used to measure compliance are reviewed. Subjective methods include patient self-reporting and physician estimates; these methods are generally inexpensive but have been shown to be unreliable. Objective methods include pill counting, electronic monitoring devices, and data reviews; these methods vary in cost and reliability. PMID- 15364636 TI - Evidence-based pain management and palliative care in issue one for 2004 of The Cochrane Library. AB - The Cochrane Library of Systematic Reviews is published quarterly. Issue one for 2004 of the library was published in February 2004. This issue contains 3,329 reviews and protocols of which 1,921 are fully published reviews. The trials database now stands at over 400,000 records with an additional 4,427 one-page summaries of non- Cochrane reviews in the NHS database of reviews of effectiveness (DARE). This version of the Library contains the results of an extensive search for RCTs on EMBASE. The latest library contains 84 new reviews, seven are considered relevant to practitioners in pain and palliative care. References are published in the same format as the citation for Cochrane reviews. PMID- 15364637 TI - Dietary supplements containing ephedrine alkaloids adulterated because they present an unreasonable risk; final rule. AB - The Food and Drug Administration issued a final regulation declaring dietary supplements containing ephedrine alkaloids adulterated under the Federal Food, Drug, and Cosmetic Act (the act) because they present an unreasonable risk of illness or injury under the conditions of use recommended or suggested in labeling, or if no conditions of use are suggested or recommended in labeling, under ordinary conditions of use. The agency took this action based upon the wellknown pharmacology of ephedrine alkaloids, the peer-reviewed scientific literature on the effects of ephedrine alkaloids, and the adverse events reported to have occurred in individuals following consumption of dietary supplements containing ephedrine alkaloids. The final Rule was published in the Federal Register, Volume 69, Number 28, Pages 6787-6854 on February 11, 2004. This rule that became effective on April 12, 2004 is now included in the Code of Federal Regulations (21CFR Part 119). This report includes the Table of Contents of the full Federal Register publication and selected sections and tables from that publication. PMID- 15364638 TI - OxyContin abuse and diversion and efforts to address the problem: highlights of a government report. AB - Representative Frank R. Wolf, Chairman of the U. S. House of Representatives Subcommittee on Commerce, Justice, State, and the Judiciary, Committee on Appropriations; and Representative James C. Greenwood, Chairman of the Subcommittee on Oversight and Investigations, Committee on Energy and Commerce, requested that the U.S. Government Accounting Office (GAO) investigate OxyContin Abuse and Diversion. The GAO is the audit, evaluation and investigative arm of Congress. On December 23, 2003, the GAO submitted a 57 page report to the Representatives describing the outcomes of the investigation. The full report includes appendices that describe the scope and methodology of the investigation, summaries of FDA changes to the original approved OxyContin label, databases used to monitor abuse and diversion of OxyContin and oxycodone, and comments from the FDA and DEA. This special report is an abstract of the GAO report and includes the conclusions and recommendations for action form the full report. PMID- 15364640 TI - Antioxidant activity of the aqueous extracts of spicy food additives--evaluation and comparison with ascorbic acid in in-vitro systems. AB - The antioxidant activity of the aqueous extracts of five umbelliferous fruits- caraway (Carum carvi), coriander (Coriandrum sativum), cumin (Cuminum cyminum), dill (Anethum graveolens) and fennel (Foeniculum vulgare)--were investigated in comparison with the known antioxidant ascorbic acid in in vitro studies. The amount of aqueous extract of these five umbelliferous fruits and ascorbic acid needed for 50% scavenging of superoxide radicals was found to be 105 microg (caraway), 370 microg (coriander), 220 microg (cumin), 190 microg (dill), 205 microg (fennel) and 260 microg (ascorbic acid). The amount needed for 50% inhibition of lipid peroxide was 2100 microg (caraway), 4500 microg (coriander), 4300 microg (cumin), 3100 microg (dill), 4600 microg (fennel) and 5000 microg (ascorbic acid). The quantity needed for 50% inhibition of hydroxyl radicals was 1150 microg (caraway), 1250 microg (coriander), 470 microg (cumin), 575 microg (dill), 700 microg (fennel) and 4500 microg (ascorbic acid). The daily use of the above fruits in various forms is very common in India and the present study revealed strong antioxidant activity of their extracts that was superior to known antioxidant ascorbic acid and indicate their intake may be beneficial as food additives. PMID- 15364641 TI - Inhibition of COX isoforms by nutraceuticals. AB - Humans have two isoforms of Prostaglandin H Synthase or cyclooxygenase: COX-1 and COX-2. COX-1 is cytoprotective. COX-2 inhibitors reduce inflammation without the risk of ulceration and kidney damage. The ideal nutraceutical would inhibit COX-2 synthesis while preserving COX-1 synthesis. The hypothesis for this research was that COX inhibitors would fall primarily into three categories: COX-2 specific inhibition, non-specific inhibition (COX-1 and COX-2), and minimal inhibition. The human Cayman COX inhibitor screening assay was used to determine the inhibitory concentration 50 (IC50) of COX-1/ COX-2 activity of each nutraceutical. The assay was run, in duplicate, with three concentrations of a suspected inhibitor, a standard curve of eight concentrations, a non-specific binding sample, and a maximum binding sample. The inhibition and concentration of each sample was then put on a multiple regression best-fit line and the IC50 determined. For comparison, ibuprofen, rofecoxib, naproxen, and indomethacin were used. Positive results were seen for ipriflavone, resveratrol, MSV-60, amentoflavone, ruscus extract and notoginseng. Glucosamine, nexrutine, and berberine did not inhibit either isoform. PMID- 15364642 TI - An open study to assess the safety and efficacy of Aesculus hippocastanum tablets (Aesculaforce 50mg) in the treatment of chronic venous insufficiency. AB - An open study was carried out to assess, primarily, the safety and tolerability of Aesculus hippocastanum in the treatment of CVI. Patients underwent 8 consecutive weeks of treatment and were asked to take one 50 mg Aesculus hippocastanum tablet, twice daily. In total, 91 adverse events were reported, of which only 4 were rated as probably related to the study drug. Patients judged the tolerability of the study medication in the majority of the cases at visits 2 and 3 (90 and 95%, respectively) to be "good" or "fairly good." Only 2 patients rated tolerability as poor at visit 3. For each of the symptoms investigated the difference in the median value between baseline and visit 3 was found to be statistically significant and both the ankle and lower leg circumference decreased. The PPG measurements were rejected after analysis since validation measurements carried out after the trial showed that the PPG technique had an internal error of around 30%. Nevertheless, the majority of patients rated efficacy to be "very good" or "good," with only 10 patients reporting no effect by the end of the study. The results of this study indicate that Aesculaforce 50 mg tablets are a safe, well-tolerated and efficacious treatment for Widmer stage I and II CVI. PMID- 15364643 TI - Correlation of cytokine and mobility in mice with arthritis and during therapy with Swertia chirayita. AB - We attempt to derive a correlation between pro-inflammatory cytokines and mobility in arthritic mice and after treatment with Swertia chirayita plant extract. Tumor necrosis factor-a (TNF-a), interleukin-1alpha (IL-1alpha), interleukin-6 (IL-6) and interleukin-10 (IL-10) were measured by solid phase sandwich ELISA and the different parameters of mobility [distance traveled (DT), resting time (RT), ambulatory time (AT) and stereotypic time (ST)] was monitored and recorded by an Auto track system. Joint circumference of the right hind limb was monitored using a Vernier caliper. Our preliminary findings revealed that DT and RT correlated well with IL-1alpha, TNF-alpha and IL-10 levels in the joint homogenates of arthritic mice and after therapy with Swertia chirayita extracts. AT and ST showed poor correlation with the cytokines. Thus we demonstrate here the anti-inflammatory property of Swertia chirayita and its usage in the prevention of arthritis in the broader aspect in the management of associated cytokines. PMID- 15364644 TI - Herbal medicine course within pharmacy curriculum. AB - Due to the growing use of herbs and dietary supplements by consumers, pharmacists now need a basic knowledge of this topic for their professional practices. To meet this need, an elective course on herbs and dietary supplements is being taught at the Massachusetts College of Pharmacy and Health Science. The objective of the course is to teach students basic phytopharmacology, counseling skills on herbal medicine, evidence- based resources, manufacturing practices, and dietary regulations. A review of over 50 herbs touches on clinical use, efficacy, safety, and drug interactions. This paper reviews the course curriculum, resources and exercises used throughout the course. PMID- 15364646 TI - Lavender (Lavandula angustifolia Miller). AB - An evidence-based systematic review including scientific evidence, expert opinion, folkloric precedent, history, pharmacology, kinetics/dynamics, interactions, adverse effects, toxicology, and dosing. PMID- 15364647 TI - Angel: post-implementation evaluation at the Indiana University School of Medicine. AB - With continued adoption of course management systems, librarians are assuming new roles and challenges with regard to providing information resources and library services in a digital environment. For approximately five years, the Indiana University School of Medicine has employed Angel as its online teaching environment, and the Ruth Lilly Medical Library has been actively involved in the adoption of Angel. As the implementation phase was completed, a post implementation evaluation was due. The authors conducted an evaluation of the Angel system to evaluate utilization level, content types, and library resources/services integration. This article provides the findings of the post implementation Angel Courses Evaluation. PMID- 15364648 TI - Cancelled requests:a study of interlibrary loan requests for items held in a medical library collection. AB - This paper presents the results of a study to document and analyze a sample of interlibrary loan requests received from library patrons of an academic medical library for items that are actually held in the collection of the library. This category is referred to as cancelled requests. The purpose of the study was to attempt to measure the degree to which library patrons are not making the best possible use of the collection and understand why this was taking place. Suggestions are presented for steps to lower the cancellation rate. PMID- 15364649 TI - Retrieval comparison of EndNote to search MEDLINE (Ovid and PubMed) versus searching them directly. AB - Using EndNote version 7.0, the authors tested the search capabilities of the EndNote search engine for retrieving citations from MEDLINE for importation into EndNote, a citation management software package. Ovid MEDLINE and PubMed were selected for the comparison. Several searches were performed on Ovid MEDLINE and PubMed using EndNote as the search engine, and the same searches were run on both Ovid and PubMed directly. Findings indicate that it is preferable to search MEDLINE directly rather than using EndNote. The publishers of EndNote do warn its users about the limitations of their product as a search engine when searching external databases. In this article, the limitations of EndNote as a search engine for searching MEDLINE were explored as related to MeSH, non-MeSH, citation verification, and author searching. PMID- 15364650 TI - Making sense of central nervous system infections at the reference desk. AB - This paper discusses concepts and terminology of some aspects of infections of the central nervous system as it relates to medical reference work. Details of anatomic, biochemical, and pathologic processes are not discussed. Specific terminology involved in this area will be reviewed in order to help ensure a good approach to developing prudent strategies for database searching of the medical literature. MeSH thesauri terms are discussed and text word synonyms are presented that provide tools for thorough searching techniques. Commonly used medical jargon for this area is also explained. Examples of specific search strategies are illustrated. PMID- 15364651 TI - PIER- evidence-base medicine from ACP. AB - PIER has been available directly to physicians from the American College of Physicians (ACP) for some time, but it has only recently become available to libraries via the STAT!Ref suite of electronic resources. PIER is designed specifically for the practicing internist, and it offers a uniform search structure that is meant to be searched the way physicians work through their medical cases, going from diagnosis to treatment. All of the information content in PIER is evidence-based and consists of clinical practice guidelines and recommendations. PMID- 15364652 TI - Infectious diseases on the World Wide Web. AB - Infectious diseases are a critically important topic in medicine. Although several of the most dangerous infectious diseases have been controlled or eliminated by modern medicine, many others continue to plague mankind. Old pathogens reappear, sometimes in drug-resistant forms, and new diseases such as SARS have emerged. The World Wide Web provides a plethora of information on infectious diseases; unfortunately, not all of it is reliable. This article lists the best Web sites on infectious diseases to help health professionals and consumers find current, accurate information on the topic. PMID- 15364653 TI - The business of negotiating for hospital librarians. AB - Although many hospital librarians may find it difficult, negotiating with vendors has become a basic skill of library acquisitions. This article reports the results of a non-scientific questionnaire administered to hospital librarians and vendors attending a chapter meeting of the Medical Library Association in 2003. The answers revealed that vendors regard libraries as businesses, and while admitting that the role is often uncomfortable for them, librarians acknowledged that negotiating skills have become an important aspect of their jobs. Questions to help guide librarians through the negotiation process are provided in the Appendix. PMID- 15364654 TI - Building a bioinformatics community of practice through library education programs. AB - This paper addresses the following questions:What makes the community of practice concept an intriguing framework for developing library services for bioinformatics? What is the campus context and setting? What has been the Health Sciences Library's role in bioinformatics at the University of North Carolina (UNC) Chapel Hill? What are the Health Sciences Library's goals? What services are currently offered? How will these services be evaluated and developed? How can libraries demonstrate their value? Providing library services for an emerging community such as bioinformatics and computational biology presents special challenges for libraries including understanding needs, defining and communicating the library's role, building relationships within the community, preparing staff, and securing funding. Like many academic health sciences libraries, the University of North Carolina (UNC) at Chapel Hill Health Sciences Library is addressing these challenges in the context of its overall mission and goals. PMID- 15364657 TI - Intergenerational service-learning across levels and disciplines:"one size (does not) fit all". AB - Service-learning is increasingly becoming an avenue used in a variety of disciplines to provide students with real-life experience regarding aging and older adults. Developing and implementing meaningful service-learning experiences, however, requires consideration both of the discipline-specific content to be addressed, as well as the varied backgrounds (educational and personal) and levels (freshman to graduate) of the students who will be participating. Intergenerational service-learning is not a one-size-fits-all endeavor. Drawing on the experiences of intergenerational service-learning programs at three institutions (representing three disciplines-Gerontology, Nursing, and Occupational Therapy), this paper focuses on the discipline-specific challenges of building a quality intergenerational service-learning program, as well as the associated challenges of developing appropriate intergenerational service-learning experiences for a range of student levels with varied backgrounds. PMID- 15364658 TI - Service learning at dementia care programs: an orientation and training program. AB - The present paper describes a project that addresses the unique challenge service learners face at dementia care programs. The project was conducted in conjunction with two courses on aging that offer students a service-learning (S-L) option at a university adult day service (ADS) program that accepts service-learners from these courses. The intervention consisted of two elements: (a) an orientation and follow- up sessions in which students learned about the causes of dementia, the behavioral characteristics exhibited by persons with dementia, and the best methods of interacting effectively with cognitively impaired persons; and (b) a structured scrapbook project for service-learners to exercise their new knowledge and skills. The authors designed the project to increase students' comfort in the dementia care setting and to facilitate students' knowledge of the elderly. The following sections describe the project and its procedures, benefits, and challenges. Recommendations for continuing the project and for adapting the project to other dementia sites are provided. PMID- 15364659 TI - Service-learners at dementia care programs:an intervention for improving contact, comfort, and attitudes. AB - Service-Learning can be a rewarding and challenging experience for students. One of the rewards for students can be the connection between their course work and real life experience. However, students interacting with populations with which they have limited prior experience face unique challenges. We developed a training program designed to facilitate comfort with older adults who have dementia for service-learners in a gerontology course who were serving at an Adult Day Services (ADS) program. Students in the course completed pre- and post surveys assessing level of contact with older adults and attitudes on aging. Service-learners serving at the ADS comprised the treatment group (n = 5) while those serving at other S-L sites made up the control group (n = 11). Independent and paired sample t-tests indicated that the training program contributed to intra-individual and group differences in comfort working with older adults and with adults who are cognitively impaired. Differences in attitudes about older adults were also identified. Implications for adult development courses involving S-L at dementia care programs are discussed. PMID- 15364660 TI - Undergraduate knowledge of aging: a comparative study of biopsychosocial content. AB - This study assesses undergraduate knowledge of aging, distinguishing between types of deficits (ignorance vs. misinformation) and content areas as delineated by a biopsychosocial framework. Knowledge is examined as an outcome of taking an aging elective, while accounting for course rating and knowledge retention. A diverse body of UCLA undergraduate students who took an aging course (n = 349) and a stratified random comparison group (n = 430) took Palmore's Facts on Aging Quiz and scored 14 and 12 items correct, respectively. Undergraduates (n = 779) showed a good knowledge of aging in the psychological- cognitive domain, but consistent with previous findings, they exhibited deficits primarily in the social-demographic realm. Taking an aging course was significantly related to better overall knowledge of aging (correct) (p .78). The 12-month quit rates, validated by either saliva cotinine or spousal proxy, were 22% in the bupropion group and 28% in the placebo group (P =.31). Based on biochemical validation, 19% of the bupropion group vs 24% of the placebo group had quit smoking by 1 year (P =.36). CONCLUSIONS: In this randomized blinded trial of mostly veteran participants, the addition of a brief 7-week bupropion trial to treatment with nicotine replacement therapy and counseling did not significantly increase smoking cessation rates. PMID- 15364676 TI - Pulmonary thromboembolism in American Indians and Alaskan Natives. AB - BACKGROUND: The rate of diagnosis of deep venous thrombosis and/or pulmonary embolism (collectively, venous thromboembolism: VTE) among patients discharged from Indian Health Service hospital care from 1980 through 1996 was considerably lower than rates reported in African Americans or whites. Expansion of the national census in 1990 to include American Indians and Alaskan Natives permits a more in-depth examination of this issue. METHODS: Combined data from the National Hospital Discharge Survey (nonfederal hospitals) and the Indian Health Service (federal hospitals) from 1996 through 2001 were used to evaluate the rate of diagnosis of VTE in American Indians and Alaskan Natives. RESULTS: The diagnosis of VTE in American Indians and Alaskan Natives, based on combined data from the National Hospital Discharge Survey and the Indian Health Service was 71 per 100,000 per year compared with 155 per 100,000 per year in African Americans (P<.001) and 131 per 100,000 per year in whites (P<.001). The rate ratio comparing the rate of diagnosis of VTE in American Indians and Alaskan Natives with African Americans was 0.46 (95% confidence interval, 0.45-0.47) and comparing American Indians and Alaskan Natives with whites it was 0.54 (95% confidence interval, 0.53-0.55). CONCLUSIONS: The observed relatively low incidence of VTE in American Indians and Alaskan Natives would seem to be due to as yet undetermined genetic factors. The possibility that American Indians and Alaskan Natives have different lifestyles that affect the rate of diagnosis of VTE cannot be excluded. PMID- 15364677 TI - Assessment of the usefulness of sputum culture for diagnosis of community acquired pneumonia using the PORT predictive scoring system. AB - BACKGROUND: The usefulness of sputum culture in guiding microbiological diagnosis of community-acquired pneumonia is controversial. We evaluate and assess it using the Patients Outcome Research Team (PORT) predictive scoring system. METHODS: A cohort of 1669 patients with community-acquired pneumonia was studied. Before administering antibiotic therapy, sputum was collected and its quality evaluated. Samples were gram stained and those of good quality were assessed for a predominant morphotype (PM). Sputum cultures were processed according to standard protocols. RESULTS: A sputum sample was obtained from 983 (59%) of the 1669 patients and 532 (54%) of the samples were of good quality. There was a PM in 240 (45%) of the latter samples (ie, for 14.4% of the 1669 patients) and there was no PM in 292 (55%). Culture yielded a microorganism in 207 (86%) of the 240 samples with PM and 57 (19.5%) of the 292 samples without PM (P<.05). Rates of sputum obtained, good-quality sputum specimens, PM identification, and positive culture were not significantly different among the PORT-score groups of patients (P>.05). The sensitivity and specificity of the gram-positive diplococci identification in the sputum culture of Streptococcus pneumoniae were 60% and 97.6%, and the positive and negative predictive values were 91% and 85.3%, respectively. CONCLUSIONS: Good-quality sputum with PM could be obtained in only 14.4% of all patients. A PORT-score group in which sputum could be of greater usefulness in identifying the causative organism could not be identified. The presence of gram positive diplococci in gram-stained sputum culture was highly specific for S pneumoniae. PMID- 15364678 TI - Neurological diagnosis is more than a state of mind: diagnostic clarity and impaired consciousness. PMID- 15364679 TI - Functional imaging of severely brain-injured patients: progress, challenges, and limitations. PMID- 15364680 TI - Monotherapy in epilepsy: role of the newer antiepileptic drugs. AB - BACKGROUND: Monotherapy is the goal for pharmacological treatment of epilepsy. Well-controlled trials have established the efficacy of some of the newer antiepileptic drugs (AEDs) as monotherapy. OBJECTIVE: To review clinical data and expert opinions pertinent to the evaluation of most of the newer AEDs as monotherapy for epilepsy. DATA SOURCES: The MEDLINE database was searched for clinical trials using newer AEDs. Reference sections of review articles were manually searched to identify relevant studies not retrieved in MEDLINE. STUDY SELECTION: The resulting list of references was manually reviewed to identify monotherapy studies. RESULTS: Lamotrigine and oxcarbazepine demonstrated efficacy in randomized active-control trials in patients with newly diagnosed epilepsy and in substitution trials in patients refractory to conventional AEDs. CONCLUSION: Lamotrigine and oxcarbazepine are as effective as conventional AEDs at controlling partial seizures and are better tolerated. PMID- 15364681 TI - Migraine is associated with magnetic resonance imaging white matter abnormalities: a meta-analysis. AB - BACKGROUND: There is controversy as to whether migraine is associated with white matter abnormalities (WMAs) on magnetic resonance images. These abnormalities may be important as a risk factor for future stroke. Further, it is controversial whether any increased risk of WMAs is attributable to comorbidities such as vascular disease. METHODS: A meta-analysis of published case-control studies was undertaken to address the relationship between migraine and magnetic resonance imaging WMAs. Seven studies were identified. Data from studies reporting the incidence of magnetic resonance imaging WMAs in those with migraine and appropriate control populations were used to calculate odds ratios for WMAs in migraine for each study. A stratified meta-analysis was performed using studies that did and did not exclude subjects with disease comorbidities. RESULTS: The summary odds ratio shows that those with migraine are at increased risk for WMAs (odds ratio, 3.9 [95% confidence interval, 2.26-6.72]). The risk does not differ between studies that included subjects with comorbidities and those that did not. CONCLUSION: This meta-analysis demonstrates that subjects with migraine are at higher risk of having WMAs on magnetic resonance images than those without migraine. This increased risk is present even in younger individuals who do not have co-occurring cerebrovascular disease risk factors. Prospective studies are needed to determine whether the increased risk of stroke in migraine is mediated or foreshadowed by the presence of WMAs. PMID- 15364682 TI - Homocysteine and the brain in midadult life: evidence for an increased risk of leukoaraiosis in men. AB - BACKGROUND: High serum homocysteine (HCY) levels have been associated with thromboembolic cerebrovascular disease, but their relationship to microvascular disease is uncertain. Homocysteine also has a direct neurotoxic effect and has been linked to brain atrophy and an increased risk of Alzheimer disease. OBJECTIVE: To examine the relationship of HCY levels to brain and cognitive measures in a healthy community sample. DESIGN: Cross-sectional study. SETTING: Individuals residing in Canberra and Queanbeyan, Australia, who were participating in the longitudinal PATH Through Life Project. PARTICIPANTS: Individuals aged 60 to 64 years selected randomly from the community, 196 men and 189 women. MAIN OUTCOME MEASURES: Regression coefficients with HCY level as the putative determinant and various magnetic resonance imaging measures (brain atrophy index, ventricle-brain ratios, volume of periventricular and deep white matter hyperintensities) and cognitive measures (information processing speed, verbal memory, fine motor speed) as dependent measures. RESULTS: Homocysteine levels did not have a significant relationship with brain atrophy index or ventricle-brain ratios. High HCY levels were related to increased deep white matter hyperintensities but not periventricular white matter hyperintensities, after correcting for levels of folate, vitamin B(12), creatinine, and thyrotropin; hypertension; smoking; and diabetes, the relationship being significant only in men. Homocysteine levels were related to impairment in verbal memory and fine motor speed but not after the previously mentioned correction. CONCLUSIONS: Total HCY level is independently related to leukoaraiosis in middle aged men, and this may be functionally relevant in the form of mild cognitive impairment. The remediation of hyperhomocysteinemia should begin early in life if its deleterious effects on the brain are to be prevented. PMID- 15364683 TI - Voxel-based morphometry reveals gray matter network atrophy in refractory medial temporal lobe epilepsy. AB - BACKGROUND: Conventional volumetric studies have shown that brain structures functionally and anatomically related to the hippocampus are smaller in patients with drug-refractory medial temporal lobe epilepsy (MTLE). OBJECTIVES: To determine the extent of gray matter atrophy in the brains of patients with MTLE and to examine the pattern of atrophy. DESIGN: We performed a voxel-based morphometric study of 43 consecutive patients with unilateral drug-refractory MTLE (21 patients with right-sided MTLE and 22 patients with left-sided MTLE) whose magnetic resonance images showed signs of unilateral hippocampal atrophy. The data from the patients with MTLE were compared with the data from 49 healthy control subjects to identify differences between groups in gray matter concentration (GMC). SETTING: Academic hospital's epilepsy clinic. RESULTS: We observed that patients with left- and right-sided MTLE exhibited GMC reduction in the hippocampus ipsilateral to the seizure origin. In addition, we found GMC reduction in the ipsilateral parahippocampal and isocortical temporal regions. Patients with MTLE also showed GMC reduction in subcortical nuclei such as the thalamus and caudate, in the cerebellum, in the midbrain, and in parieto occipital regions. CONCLUSIONS: Patients with MTLE exhibit a reduction in GMC in regions outside the temporal lobe, specifically in areas that are connected to the hippocampus and parahippocampal region, suggesting an anatomical route for atrophy. PMID- 15364684 TI - Neurologic impairment 10 years after optic neuritis. AB - BACKGROUND: Participants enrolled in the Optic Neuritis Treatment Trial have been observed for more than a decade to assess the relationship between optic neuritis and the development of clinically definite multiple sclerosis. OBJECTIVE: To assess neurologic disability 10 to 12 years after an initial episode of optic neuritis. DESIGN: Longitudinal follow-up of a clinical trial. SETTING: Fourteen Optic Neuritis Treatment Trial clinical centers performed standardized neurologic examinations, including an assessment of neurologic disability. PARTICIPANTS: One hundred twenty-seven patients who had developed clinically definite multiple sclerosis. MAIN OUTCOME MEASURES: Functional Systems Scale and Expanded Disability Status Scale. RESULTS: The disability of most patients was mild, with 65% of patients having an Expanded Disability Status Scale score lower than 3.0. The degree of disability appeared to be unrelated to whether the baseline magnetic resonance imaging scan was lesion-free or showed lesions (P =.51). Among patients with baseline lesions, the degree of disability was unrelated to the number of lesions that were present on the scan (P =.14). Two patients died owing to severe multiple sclerosis, one of whom had no lesions revealed on the baseline scan. CONCLUSION: Most patients who develop clinically definite multiple sclerosis following an initial episode of optic neuritis will have a relatively benign course for at least 10 years. PMID- 15364685 TI - Long-term treatment of restless legs syndrome with dopamine agonists. AB - BACKGROUND: Controlled clinical trials robustly demonstrate the short-term efficacy of dopamine agonists (DA) for restless legs syndrome (RLS), but little is known about the long-term efficacy and long-term adverse events. Augmentation an increase in the duration, intensity, and anatomy of RLS symptoms-is commonly associated with dopaminergic treatments; however, risk factors for this troubling scenario have not been formally evaluated. OBJECTIVES: To evaluate the long-term efficacy and tolerability of DA for RLS and to evaluate factors that could predict the occurrence of augmentation. METHODS: We queried all subjects seen from 1996 to 2003 and followed up those initiated on any DA by the Baylor College of Medicine Movement Disorders Clinic, Houston, Tex. Patients with Parkinson disease, uremia, or medications that could affect RLS were excluded. Demographics, efficacy, dosing, adverse events, and augmentation were tracked across time. Statistical modeling was used to evaluate for factors that could predict augmentation. RESULTS: After eliminating all patients with RLS who had factors that could affect DA dosing or the accuracy of data, we observed 83 subjects with at least 6 months' use of DA (mean +/- SD, 39.2 +/- 20.9 months). Efficacy was maintained across time but at the expense of moderate but significant increases in doses (P<.01). Adverse events were frequent but usually mild and seldom resulted in discontinuation. Augmentation was frequent (48% of subjects) but usually modest, and it was predicted by a positive family history for RLS and especially the lack of any neuropathy on electromyographic or nerve conduction velocity tests. CONCLUSIONS: Dopamine agonists continued to effectively treat RLS without long-term adverse events but often required adjustments across time. The higher rate of augmentation in familial and nonneuropathic RLS should be considered when initiating therapy. PMID- 15364686 TI - The natural history of recurrent optic neuritis. AB - BACKGROUND: Optic neuritis (ON) may occur in isolation or may herald multiple sclerosis (MS) or neuromyelitis optica (NMO). Occasionally, ON may recur many times without intervening evidence of dissemination in space. OBJECTIVE: To define the clinical course and prognosis of patients with recurrent ON. DESIGN: Retrospective medical record review and telephone follow-up survey. SETTING: Clinic-based practice in a large tertiary referral institution. MAIN OUTCOME MEASURES: Survival analysis of conversion to MS and NMO and final visual impairment. We studied the association of clinical and demographic factors, the presence of brain lesions on magnetic resonance images, and the use of corticosteroid treatment at the time of the first ON occurrence with conversion to MS and NMO. RESULTS: We identified 1274 patients with ON between 1994 and 2000 and selected 72 (5.7%) with recurrent ON without intervening symptoms of a disseminated demyelinating condition for further analysis. The 5-year conversion rate to NMO was 12.5% and to MS, 14.4%. Among 5 patients with 2 or more lesions consistent with MS on brain magnetic resonance images, 2 (40.0%) converted to MS and none to NMO, while among 11 patients without such lesions, none converted to MS and 2 (18.2%) converted to NMO (P =.16). Conversion to MS occurred in 7 (19.4%) of 36 individuals treated for their first ON episode with corticosteroids vs 4 (44.4%) of 9 untreated individuals (P =.19). There was no difference in the conversion rate to MS between those treated with intravenous steroids (4 [16.7%] of 24) vs oral steroids (3 [25.0%] of 12) (P =.33). Conversion to NMO occurred earlier than conversion to MS (2.3 +/- 1.6 vs 5.3 +/- 4.3 years, respectively; P =.01). Women tended to convert to NMO more frequently than men (female-male ratio for NMO converters, 7:1; MS converters, 2:1; nonconverters, 2:1; P =.56), as did those with a higher annual frequency of ON episodes (NMO converters, 2.0 +/- 1.3; MS converters, 1.0 +/- 1.0; nonconverters, 0.6 +/- 0.5; P =.04). The number of ON events in the first 2 years following the first ON episode was higher in the NMO group (NMO converters, 2.4 +/- 0.9; MS converters, 1.9 +/- 1.1; nonconverters, 1.7 +/- 0.7; P =.04). The final visual impairment was greatest in the NMO group (P =.02). CONCLUSIONS: Patients with rapid succession of severe ON events are more likely to develop a generalized demyelinating disease. Patients with NMO had a worse visual outcome. PMID- 15364687 TI - European study on intravenous immunoglobulin in multiple sclerosis: results of magnetization transfer magnetic resonance imaging analysis. AB - BACKGROUND: Magnetization transfer magnetic resonance imaging (MT MRI) can provide in vivo markers reflecting the severity of multiple sclerosis-related brain damage occurring within and outside T2-visible lesions. OBJECTIVE: To investigate the effect of intravenous immunoglobulin (IVIG) treatment on the accumulation of brain damage in patients with secondary progressive multiple sclerosis (SPMS), measured using MT MRI.Design, Patients, and Intervention Seventy patients with SPMS participating in the European, multicenter, randomized, double-blind, placebo-controlled trial of IVIG in SPMS underwent brain T2-weighted and MT MRI at baseline and after 12 and 24 months. The MT MRI scans were post-processed and analyzed to obtain MT ratio values from T2-visible lesions and MT ratio histograms from the normal-appearing brain tissue (NABT). RESULTS: At baseline, a significant difference was found for NABT MT ratio histogram peak height (P =.003) between treated patients and patients receiving placebo. No significant differences between treated patients and those receiving placebo were found for any of the considered MT MRI-derived metrics in terms of treatment x time interaction. Nevertheless, over the 24-month period, the placebo patients experienced a 6.75% reduction of the NABT MT ratio histogram peak height, whereas treated patients experienced only a 0.92% reduction of the NABT MT ratio histogram peak height. CONCLUSIONS: This study did not show any statistically significant effect of IVIG on MT MRI quantities. Nevertheless, the markedly different percentage change of the NABT MT ratio histogram peak height over time between patients receiving placebo and treated patients suggests a possible role of IVIG treatment in preventing the loss of "truly" normal brain tissue in SPMS patients. PMID- 15364688 TI - Botulinum toxin type A for treating voice tremor. AB - BACKGROUND: Voice tremor, like spasmodic dysphonia and other tremor disorders, may respond to botulinum toxin type A injections. OBJECTIVE: To evaluate the safety and efficacy of botulinum toxin type A injections as treatment for voice tremor. DESIGN: A randomized study of 3 doses of botulinum toxin type A with 6 weeks of follow-up. SETTING: A single-site tertiary care center. PARTICIPANTS AND METHODS: Thirteen subjects (11 women, 2 men; mean age, 73 years) with voice tremor and no spasmodic dysphonia or head, mouth, jaw, or facial tremor were entered into this study. Patients received 1.25 U (n = 5), 2.5 U (n = 5), or 3.75 U (n = 3) of botulinum toxin type A in each vocal cord. All patients were evaluated at baseline and postinjection at weeks 2, 4, and 6. MAIN OUTCOME MEASURES: The primary outcome measure was the patient tremor rating scale, with secondary measures including patient-rated functional disability, response rating scale, independent randomized tremor ratings, and acoustical measures. RESULTS: All patients at all dose levels noted an effect from the injection. The mean time to onset of effect was 2.3 days (range, 1-7 days). For all patients combined, mean tremor severity scale scores (rated by patients on a 5-point scale) improved 1.4 points at week 2, 1.6 points at week 4, and 1.7 points at week 6. Measures of functional disability, measures of the effect of injection, independent ratings of videotaped speech, and acoustic measures of tremor also showed improvement. The main adverse effects at all doses were breathiness and dysphagia. CONCLUSION: Voice tremor improves following injections of botulinum toxin type A. PMID- 15364689 TI - Multiple myeloma invasion of the central nervous system. AB - BACKGROUND: Although neurologic manifestations often complicate the course of patients with multiple myeloma (MM), direct central nervous system invasion is rare. OBJECTIVE: To describe the neurologic symptoms and signs, imaging, cerebrospinal fluid findings, and the clinical course of patients with central nervous system myeloma invasion, all of whom had leptomeningeal myelomatosis. DESIGN AND PARTICIPANTS: Review of 23 patients with MM and leptomeningeal myelomatosis proven by malignant plasma cells in their cerebrospinal fluid. SETTING: Tertiary-care university medical center. RESULTS: Twenty-one patients had advanced-stage MM. Leptomeningeal myelomatosis was diagnosed up to 29 months (median, 13 months) after diagnosis of MM. Symptoms precipitating neurologic evaluation included manifestations of diffuse cerebral dysfunction, cranial nerve palsies, and spinal radiculopathies. Cerebrospinal fluid was abnormal in all patients, usually exhibiting pleocytosis and elevated protein content, plus positive cytologic findings. Specific magnetic resonance imaging findings suggestive of central nervous system invasion were found in 70% of the patients. These included leptomeningeal contrast enhancement and evidence of meningeal based lesions sometimes masquerading as intraparenchymal lesions. Despite aggressive systemic and local treatment, the outcome was poor, reflecting the aggressiveness of the underlying MM. CONCLUSION: Leptomeningeal myelomatosis, although rare, should be considered in patients with MM and symptoms suggestive of widespread nervous system involvement. PMID- 15364690 TI - APOE and APOC1 promoter polymorphisms and the risk of Alzheimer disease in African American and Caribbean Hispanic individuals. AB - BACKGROUND: The APOE epsilon4 allele is a genetic risk factor for Alzheimer disease (AD), though the strength of the association varies by ethnic group. Polymorphisms in regulatory sequences of APOE have also been related to AD, but the effects are inconsistent across studies. METHODS: We examined the association between AD and variants in 3 APOE promoters and in the promoter of the adjacent APOC1 gene in African American and Caribbean Hispanic individuals. Polymorphisms tested were the -491A/T, -427T/C, and -219G/T (Th1/E47cs) in the APOE promoter and the HpaI variant in the APOC1 promoter. Using standard research criteria for AD, overall odds ratios were computed and repeated stratified by presence or absence of APOE epsilon4. RESULTS: The APOC1 HpaI+ variant was associated with AD in Caribbean Hispanic individuals, but strong linkage disequilibrium with the APOE epsilon4 allele indicated that this was not an independent effect. No promoter variant in APOE or APOC1 was associated with AD before or after adjusting for age, education, sex, and multiple comparisons. Estimated haplotypes including -219G/T, APOE, and APOC1 differed significantly in Caribbean Hispanic patients and controls but not in African American participants. This effect was primarily owing to the -219G/T-APOE haplotype, but we did not detect significant allele-specific differences in promoter activity comparing reporter constructs containing the APOE -219G and -219 T alleles. CONCLUSION: These findings exclude a strong or independent influence of APOE or APOC1 promoter polymorphisms on the variation in APOE-related risk of AD in African American and Caribbean Hispanic individuals. PMID- 15364691 TI - Vestibular imbalance associated with a lesion in the nucleus prepositus hypoglossi area. AB - BACKGROUND: The nucleus prepositus hypoglossi (NPH) is known to be a neural integrator of horizontal eye movements. Although the role of the human NPH is not well known, it may also function in postural balance, in view of its anatomic connections with the vestibular nuclei and vestibulocerebellum and of lesion studies in experimental animals. OBJECTIVE: To show that the human NPH contributes to vestibular function in addition to eye movement control. DESIGN: Case series. SETTING: University hospital. Patients Six patients with small and discrete brainstem infarctions that predominantly involved the NPH region.Main Outcome Measure Findings on magnetic resonance images. RESULTS: The NPH was affected at the lower pontine level in 2 patients and at the upper medullary level in 4. In addition to gaze-evoked nystagmus, all patients had vertigo, vomiting, and postural ataxia, suggesting vestibular dysfunction. The patients typically fell contralaterally or bilaterally to the lesion side. CONCLUSION: The NPH serves a vestibular function in addition to its oculomotor control function. PMID- 15364692 TI - Pupil-sparing, painless compression of the oculomotor nerve by expanding basilar artery aneurysm: a case of ocular pseudomyasthenia. AB - BACKGROUND: Oculomotor nerve paresis may have relatively benign but also life threatening causes. Distinguishing between these is of great clinical importance. OBJECTIVE: To reveal a potential pitfall of the clinical evaluation of oculomotor nerve paresis. PATIENT: Single case observation. RESULTS: A 56-year-old man had fluctuating diplopia and fatigable ptosis, promptly relieved by intravenous edrophonium, leading to the diagnosis of ocular myasthenia gravis. His pupillary function was intact. A few days after the initial diagnosis, he suffered a subarachnoid hemorrhage secondary to the rupture of a basilar artery aneurysm. His ocular symptoms were related to aneurysmal oculomotor nerve compression. CONCLUSION: Patients with oculomotor nerve dysfunction need more detailed evaluation because the underlying cause cannot be safely determined on a clinical basis. PMID- 15364693 TI - Isolated relative afferent pupillary defect secondary to contralateral midbrain compression. AB - BACKGROUND: Relative afferent pupillary defects are typically related to ipsilateral lesions within the anterior visual pathways. OBJECTIVE: To describe a patient who had a workup for headache and was found to have an isolated left relative afferent pupillary defect without any other neurological findings. DESIGN: We review the neuroanatomy of the pupillary light reflex pathway and emphasize the nasotemporal bias of decussating fiber projections, which accounts for the relative afferent pupillary defect contralateral to the described lesion. Result Magnetic resonance imaging of the brain revealed a pineal tumor compressing the right rostral midbrain. CONCLUSION: While rare, a relative afferent pupillary defect can occasionally occur secondary to lesions in the postchiasmal pathways. In these circumstances, the pupillary defect will be observed to be contralateral to the side of the lesion. PMID- 15364694 TI - Prominent cerebellar symptoms with unusual magnetic resonance imaging findings in acquired hepatocerebral degeneration. AB - BACKGROUND: Cerebellar lesions revealed by abnormal signals on magnetic resonance images are extremely rare in acquired hepatocerebral degeneration (AHCD). OBJECTIVE: To report a case of AHCD with prominent cerebellar findings both clinically and radiologically. DESIGN AND SETTING: Case report and tertiary-care hospital. PATIENT: A 46-year-old man complained of progressive speech difficulties of 5 months' duration. Two years earlier, he had been diagnosed as having cirrhosis of the liver caused by alcoholism and hepatitis B virus infection. RESULTS: The patient had progressive ataxic dysarthria and limb and gait ataxia as manifestations of AHCD. Magnetic resonance imaging of the brain revealed distinctive symmetrical T2 high-signal intensities in the bilateral cerebellar hemispheres and brachium pontis, which were consistent with his neurologic deficits. Simultaneously, high T1 signals in the bilateral pallidum and ventral midbrain were noted, which are typical manifestations of AHCD. Follow up magnetic resonance imaging 3 months later showed the same cerebellar signs and abnormal signals. CONCLUSIONS: The cerebellar cortex and middle cerebellar peduncle are considered highly vulnerable structures to metabolic insults in liver disease. Findings from our patient suggest that dominant cerebellar deficits with compatible T2 high-signal lesions are another type of clinical manifestation in AHCD. PMID- 15364695 TI - Isolated gait ataxia due to cerebellar vermis infarct. PMID- 15364696 TI - Magnetic resonance imaging reveals selective vulnerability of the cerebellum and basal ganglia in malignant hyperthermia. PMID- 15364697 TI - Digital rendition of visual migraines. PMID- 15364698 TI - Tay-Sachs disease. PMID- 15364699 TI - Disease course of Charcot-Marie-Tooth disease type 2 and comorbidity. PMID- 15364700 TI - Allan-Herndon-Dudley syndrome: should the locus for this hereditary spastic paraplegia be designated SPG 22? PMID- 15364701 TI - Action myoclonus-renal failure syndrome: characterization of a unique cerebro renal disorder. AB - Action myoclonus-renal failure syndrome (AMRF) is a distinctive form of progressive myoclonus epilepsy associated with renal dysfunction. The syndrome was not recognized prior to the advent of dialysis and renal transplantation because of its rapidly fatal course if renal failure is untreated. The first and only description of AMRF was in four French Canadian patients in three families (Andermann et al., 1986). We now describe 15 individuals with AMRF from five countries, including a follow-up of the four French Canadian patients, allowing a more complete characterization of this disease. Our 15 patients with AMRF belong to nine different families. Segregation analyses were compatible with autosomal recessive inheritance. In addition, our findings show that AMRF can present with either renal or neurological features. Tremor (onset 17-26 years, mean 19.8 years, median 19 years) and progressively disabling action myoclonus (onset 14-29 years, mean 21.7 years, median 21 years), with infrequent generalized seizures (onset 20-28 years, mean 22.7 years, median 22 years) and cerebellar features are characteristic. Proteinuria, detected between ages 9 and 30 years in all cases, progressed to renal failure in 12 out of 15 patients within 0-8 years after proteinuria detection. Brain autopsy in two patients revealed extraneuronal pigment accumulation. Renal biopsies showed collapsing glomerulopathy, a severe variant of focal glomerulosclerosis. This study extends the AMRF phenotype, and demonstrates a more extensive ethnic and geographic distribution of a syndrome originally believed to be confined to individuals of French Canadian ancestry. The independent progression of neurological and renal disorders in AMRF suggests a unitary molecular lesion with pleiotropic effects. Our results demonstrate that the renal lesion in AMRF is a recessive form of collapsing glomerulopathy. Genes identified for focal segmental glomerulosclerosis and involved with the function of the glomerular basement membrane and related proteins are thus good candidates. Treatment can improve quality of life and extend the lifespan of these patients. Dialysis and renal transplantation are effective for the renal but not the neurological features, which continue to progress even in the presence of normalized renal function; the latter can be managed with anti myoclonic and anti-epileptic drugs. PMID- 15364702 TI - Long-term prognosis and causes of death in CADASIL: a retrospective study in 411 patients. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a hereditary angiopathy caused by mutations in the NOTCH3 gene. The clinical course is highly variable. Little is known about the long-term prognosis and the causes of death in CADASIL patients. Likewise, the impact of gender and NOTCH3 genotype on disease progression remains largely unexplored. We identified 411 subjects (196 men, 215 women) with a definite diagnosis of CADASIL. Age at onset for stroke, immobilization and death as well as the causes of death and clinical status at onset of the cause of death were determined systematically. Weibull regression models were used to calculate times to event, with gender and NOTCH3 genotype as covariates. At the time of the study, 73 patients had died. The median age at onset for stroke was 50.7 years [95% confidence interval (CI) = 48.2-53.1 years] in men and 52.5 years (95% CI = 50.0-54.9 years) in women (P = n.s.). The median ages at onset for inability to walk without assistance [men 58.9 years (95% CI = 56.6-61.3 years); women 62.1 years (59.7-64.4 years)], bedriddenness [men 62.1 years (59.6-64.7 years), women 66.5 years (63.9-69.1 years); and death [men 64.6 years (61.7-67.6 years); women 70.7 years (67.6-73.9 years)] were significantly lower in men than in women (all P < or = 0.01). The median survival time of men was significantly shorter than expected from German life tables (64.6 versus 69.3 years, P = 0.01). In contrast, the median survival time of women was not significantly reduced (70.7 versus 72.2 years). The C117F mutation was associated with a lower age at death and the C174Y mutation with a lower age at onset for stroke, immobilization and death (adjusted P values <0.05). At onset of the cause of death, 78% of the subjects were completely dependent. Sixty-three per cent were confined to bed. Pneumonia was the most frequent cause of death (38%), followed by sudden unexpected death (26%) and asphyxia (12%). We conclude that male sex is a risk factor for early immobilization and death in CADASIL. Our findings suggest possible genotype phenotype correlations with regard to disease progression. The data presented may serve as source material for counselling CADASIL patients and for designing future interventional trials. PMID- 15364703 TI - Interventions for basal cell carcinoma of the skin: systematic review. AB - OBJECTIVES: To assess the effects of treatments for basal cell carcinoma. METHODS: Systematic review of randomised controlled trials. MAIN OUTCOME MEASURE: Recurrence of basal cell carcinoma at three years or beyond, assessed clinically. STUDIES REVIEWED: Randomised controlled trials of interventions for histologically confirmed basal cell carcinoma (published and unpublished material; no language restrictions). RESULTS: 25 studies were identified, covering seven therapeutic categories. Only one study of surgical excision versus radiotherapy contained primary outcome data, which showed significantly more persistent tumours and recurrences in the radiotherapy group compared with surgery (odds ratio 0.09, 95% confidence interval 0.01 to 0.67). One study compared cryotherapy with surgery, with inconclusive results at one year. In a comparison of radiotherapy with cryotherapy, significantly more recurrences occurred at one year in the cryotherapy group. Preliminary studies suggest a short term success rate of 87-88% for imiquimod cream in the treatment of superficial basal cell carcinoma, although this cream has not been compared with surgery. No consistent evidence was found for the other treatment modalities. CONCLUSIONS: Little good quality research has been done on the treatments used for the most common cancer in humans. Most trials have included only people with basal cell carcinoma occurring at low risk sites. Only one trial measured recurrence at four years; recurrence rates at one year should be interpreted with caution. Surgery and radiotherapy seem to be the most effective treatments; surgery showed the lowest failure rates. Other treatments might have some use but need to be compared with surgery. PMID- 15364704 TI - Relationship between tear volume and tear meniscus curvature. AB - OBJECTIVE: To investigate the relationship between tear volume and tear meniscus curvature by means of the video meniscometer. METHODS: Eleven eyes of 11 patients with severe dry eye were studied (all female; mean +/- SD age, 66.2 +/- 7.7 years; 7 left eyes and 4 right eyes), each of whose puncta had been therapeutically occluded. Four instillations of balanced salt solution were given at intervals of 1 minute in each studied eye, with the concentration increasing in 5-microL steps from 5 to 20 microL. Before and after the instillation of balanced salt solution, tear meniscus changes were recorded by video meniscometer and radius of the meniscus was calculated from the printed images by means of the concave mirror formula. RESULTS: The mean radius of the meniscus increased linearly with increased drop volume (r(2) = 0.65, P<.001), with mean +/- SD radius values of 0.24 +/- 0.08 mm at baseline and 0.48 +/- 0.13, 0.62 +/- 0.13, 0.84 +/- 0.26, and 1.00 +/- 0.32 mm after separate instillations of 5, 10, 15, and 20 microL of balanced salt solution, respectively. For each subject, a significantly different slope defining the relationship between instilled volume and meniscus radius was seen (0.016-0.063 mm/ microL; mean +/- SD, 0.039 +/- 0.015 mm/ microL), which was thought to depend on the difference in capacity of the fluid reservoir over the ocular surface. CONCLUSIONS: This study confirmed that the volume of instilled eyedrops is linearly related to the resulting radius of the tear meniscus, suggesting that this radius is a useful measure in monitoring the tear volume. This is likely to have implications both for dry eye diagnosis and for confirming the efficacy of punctal occlusion in this condition. PMID- 15364705 TI - Clinical significance of central corneal thickness in the management of glaucoma. AB - OBJECTIVE: To evaluate the effect of central corneal thickness determination on the clinical management of patients with glaucoma and glaucoma suspect. METHODS: A cross-sectional retrospective study was performed on 188 consecutive patients. Mean ultrasound pachymetry measurements of central corneal thickness and corresponding Goldmann applanation tonometry measurements were obtained. Intraocular pressures (IOPs) were corrected using linear and mathematical (Orssengo-Pye) algorithms. Measurement-significant outcomes were defined as an IOP adjustment of 1.5 mm Hg or greater and outcomes-significant results as an IOP adjustment of 3.0 mm Hg or greater. Changes in therapy such as the use of eyedrops and addition or cancellation of laser therapy or surgery were then noted for those individuals with measurement- or outcomes-significant changes. RESULTS: Using the linear correction scale, 105 (55.9%) of 188 patients had at least a measurement-significant adjustment in their IOP measurements: 67 (35.6%) had adjustments between 1.5 and 3.0 mm Hg, while 38 (20.2%) had an outcomes significant IOP adjustment (> or =3.0 mm Hg). Among the 188 patients, 16 (8.5%) had a change in eyedrop therapy, 4 (2.1%) had a change regarding laser therapy, and 6 (3.2%) had a change in the decision regarding glaucoma surgery. Using the exponential correction (Orssengo-Pye) scale, similar percentages were obtained. CONCLUSION: Pachymetry-measured central corneal thickness has a significant effect on the clinical management of patients with glaucoma and glaucoma suspect. PMID- 15364706 TI - A method for comparing electrophysiological, psychophysical, and structural measures of glaucomatous damage. AB - OBJECTIVE: To develop a method for comparing multifocal visual-evoked potential (mfVEP) responses and behaviorally determined visual fields with structural measures of the optic nerve head. METHODS: Humphrey 24-2 visual fields and mfVEPs were obtained from each eye of 20 patients with open-angle glaucoma. Monocular and interocular analyses were performed to identify locations with abnormal mfVEP responses. Optic discs were assessed with a confocal scanning laser ophthalmoscope (Heidelberg Retina Tomograph II). The image of the optic nerve head was divided into 6 sectors. The rim and disc area measurements for each sector were compared with those in a normal database using Moorfields regression analysis. The optic nerve head measurements for the 6 sectors were related to the Humphrey visual field locations and the 60 sectors of the mfVEP display. RESULTS: Of 240 sectors tested (40 eyes x 6 sectors), 18.8% on Humphrey visual field, 22.1% on mfVEP, and 10.8% on confocal scanning laser ophthalmoscopic testing were significantly different from those of control subjects. There were no significant deficits in 165 sectors. There was agreement for 86.7% of the sectors when the Humphrey visual field and mfVEP results were compared. The confocal scanning laser ophthalmoscopic results were in agreement for 84.6% of these sectors. CONCLUSIONS: The method used allows for a comparison among measures of visual function and a structural measure of the optic nerve head. In general, the results of the functional and structural measures showed agreement; however, there were clear examples of disagreements that merit further study. PMID- 15364707 TI - Dynamic contour tonometry: a comparative study on human cadaver eyes. AB - OBJECTIVE: To compare intraocular pressure measurements obtained by recently introduced dynamic contour tonometry (DCT), Goldmann applanation tonometry (GAT), pneumatonometry (PTG), and intracameral manometry in human cadaver eyes. METHODS: Sixteen freshly enucleated human cadaver eyes were deepithelialized and dehydrated with dextran. A tube was placed in the anterior chamber and connected to a transducer and to a bottle system filled with balanced salt solution. The pressure in the eye was then altered between 5 mm Hg and 58 mm Hg. Intraocular pressure measurements were obtained with DCT, GAT, and PTG at each manometric pressure reading. RESULTS: On average, DCT values measured 0.58 +/- 0.70 mm Hg higher than real intracameral pressure. The GAT and PGT showed consistently lower values, -4.01 +/- 1.76 mm Hg and -5.09 +/- 2.61 mm Hg, respectively. At all bottle heights, DCT values were significantly closer to the reference pressure than GAT and PTG (P<.001). CONCLUSIONS: Measurement with DCT provides IOP values significantly closer to true manometric levels than either GAT or PTG. Further studies are warranted to determine its reliability in patients and the effect of corneal thickness. PMID- 15364708 TI - Clinical trial of docosahexaenoic acid in patients with retinitis pigmentosa receiving vitamin A treatment. AB - OBJECTIVE: To determine whether a therapeutic dose of docosahexaenoic acid (DHA), an omega-3 fatty acid, will slow the course of retinal degeneration in adult patients with retinitis pigmentosa who are also receiving vitamin A. DESIGN: Randomized, controlled, double-masked trial of 221 patients, aged 18 to 55 years, evaluated over a 4-year interval. Patients were given either 1200 mg/d of docosahexaenoic acid or control capsules. All were given 15 000 IU/d of vitamin A (given as retinyl palmitate). Randomization considered genetic type and baseline dietary omega-3 fatty acid intake. MAIN OUTCOME MEASURES: The primary outcome measure was the total point score for the 30-2 program of the Humphrey field analyzer; secondary outcome measures were the total point score for the 30-2 and 30/60-1 programs combined, 30-Hz electroretinogram amplitude, and Early Treatment Diabetic Rentinopathy Study visual acuity. RESULTS: No significant differences in decline in ocular function were found between the docosahexaenoic acid plus vitamin A (DHA + A) group and control plus vitamin A (control + A) group over a 4 year interval among all 221 randomized patients or among the 208 patients who completed all 4 follow-up visits. The mean annual rate of loss of sensitivity for the Humphrey Field Analyzer 30-2 program was 37 dB for the DHA + A group and 38 dB for the control + A group (P =.88). For the Humphrey Field Analyzer 30-2 and 30/60-1 programs combined, the mean annual rates of loss of field sensitivity were 57 dB for the DHA + A group and 60 dB (P =.73) for control + A group. No toxic adverse effects were observed. No significant differences by treatment group assignment were observed within genetic types or within the category of baseline omega-3 fatty acid intake. CONCLUSION: In patients assigned to receive 15 000 IU/d of vitamin A, this randomized trial showed that 1200 mg/d of docosahexaenoic acid supplementation over a 4-year interval did not, on average, slow the course of disease in patients with retinitis pigmentosa. PMID- 15364709 TI - Further evaluation of docosahexaenoic acid in patients with retinitis pigmentosa receiving vitamin A treatment: subgroup analyses. AB - OBJECTIVE: To determine whether docosahexaenoic acid will slow the course of retinal degeneration in subgroups of patients with retinitis pigmentosa who are receiving vitamin A. DESIGN: A cohort of 208 patients with retinitis pigmentosa, aged 18 to 55 years, were randomly assigned to 1200 mg of docosahexaenoic acid plus 15 000 IU/d of vitamin A given as retinyl palmitate (DHA + A group) or control fatty acid plus 15 000 IU/d of vitamin A (control + A group) and followed up over 4 years. Seventy percent of the patients in each group were taking vitamin A, 15 000 IU/d, prior to entry. We compared rates of decline in ocular function in the DHA + A vs control + A groups among the subgroups defined by use or nonuse of vitamin A prior to entry. We also determined whether decline in ocular function was related to red blood cell phosphatidylethanolamine docosahexaenoic acid level, dietary omega-3 fatty acid intake, or duration of vitamin A use. Main outcome measures were Humphrey Field Analyzer visual field sensitivity, 30-Hz electroretinogram amplitude, and visual acuity. RESULTS: Among patients not taking vitamin A prior to entry, those in the DHA + A group had a slower decline in field sensitivity and electroretinogram amplitude than those in the control + A group over the first 2 years (P =.01 and P =.03, respectively); these differences were not observed in years 3 and 4 of follow-up or among patients taking vitamin A prior to entry. In the entire cohort, red blood cell phosphatidylethanolamine docosahexaenoic acid level was inversely related to rate of decline in total field sensitivity over 4 years (test for trend, P =.05). This was particularly evident over the first 2 years among those not on vitamin A prior to entry (test for trend, P =.003). In the entire control + A group, dietary omega-3 fatty acid intake was inversely related to loss of total field sensitivity over 4 years (intake, <0.20 vs > or =0.20 g/d; P =.02). The duration of vitamin A supplementation prior to entry was inversely related to rate of decline in electroretinogram amplitude (P =.008). CONCLUSIONS: For patients with retinitis pigmentosa beginning vitamin A therapy, addition of docosahexaenoic acid, 1200 mg/d, slowed the course of disease for 2 years. Among patients on vitamin A for at least 2 years, a diet rich in omega-3 fatty acids (> or =0.20 g/d) slowed the decline in visual field sensitivity. PMID- 15364710 TI - Outcome following initial external beam radiotherapy in patients with Reese Ellsworth group Vb retinoblastoma. AB - OBJECTIVE: To describe patient and ocular outcomes following initial treatment with external beam radiotherapy (EBT) in eyes with Reese-Ellsworth group Vb retinoblastoma. METHODS: Retrospective case series (from January 1, 1979, to February 28, 2002). The Kaplan-Meier method was used to analyze survival (ocular and patient) and incidence (second cancer) data. RESULTS: Two hundred forty-three patients with 1 or more Reese-Ellsworth group Vb eyes were identified. Of 284 group Vb eyes, 63 (22.2%) initially received EBT, vs 172 (60.6%) that were initially enucleated. Of the 63 radiated group Vb eyes, 31 (49.2%) had no further tumor growth, 26 (41.3%) developed a recurrence, and 8 (12.7%) developed a new tumor. Of the 63 radiated group Vb eyes, 33 (52.4%) developed ocular complications. The ocular survival rate of radiated group Vb eyes was 81.4% at 1 year and 53.4% at 10 years. Twenty-eight radiated group Vb eyes survived to the last follow-up with visual acuity information. Thirteen patients developed second cancers, 11 in the field of radiation. The probability of developing a second cancer following initial EBT for group Vb disease in patients with bilateral disease was 29.7% by 10 years after diagnosis. Survival from second cancers in patients with bilateral disease initially receiving EBT for group Vb disease was 93.6% at 5 years and 52.6% at 18(1/4) years. No patient with unilateral disease developed a second cancer. Deaths due to metastatic retinoblastoma were uncommon. CONCLUSIONS: To our knowledge, this is the first study focusing exclusively on group Vb eyes treated initially with EBT, most of which were salvaged with vision. Outcome data provided herein are clinically relevant when choosing treatment options for advanced intraocular retinoblastoma. PMID- 15364711 TI - Visual fields in retinoblastoma survivors. AB - OBJECTIVE: To describe the visual field defects in retinoblastoma survivors and relate those defects to characteristics such as tumor size, tumor location, and treatment modality. METHODS: Thirty-one patients treated for retinoblastoma were included in this study. Humphrey visual fields were determined in 33 eyes. RESULTS: Twenty-seven patients (29 eyes, 68 tumors) had sufficient diagnosis and treatment data available for further analysis. Twenty-six of the 27 patients had both absolute and relative visual field defects. Four types of visual field defects were observed and correlated with location of the tumor and therapy to the individual tumors: (1) no residual defect, (2) absolute scotoma, (3) arcuate and sector scotoma, and (4) "pseudo"-visual field defects caused by relative enophthalmos resulting from radiation. CONCLUSIONS: Patients with retinoblastoma demonstrate a variety of long-term visual field defects after treatment for their intraocular disease. Characteristics that determine the size and type of defects are tumor size, tumor location, and treatment method. PMID- 15364712 TI - Penetrating keratoplasty performed by residents. AB - OBJECTIVE: To report the results of penetrating keratoplasty performed by residents. METHOD: A retrospective medical record review of all patients undergoing penetrating keratoplasty performed by residents at our institution from April 1998 to April 2002. RESULTS: Forty penetrating keratoplasty procedures were performed by 8 residents. The most common indication was keratoconus (17 eyes [43%]), followed by corneal scarring (14 eyes [35%]). Mean preoperative best corrected visual acuity was 20/250. No intraoperative complications were reported. Mean follow-up time was 15 months. Postoperatively, mean best-corrected visual acuity was 20/40, mean postoperative astigmatism was 3.4 +/- 2.1 diopters, and graft survival was 92.5%. Postoperative complications included elevated intraocular pressure, wound dehiscence, and endophthalmitis. MAIN OUTCOME MEASURES: Best-corrected visual acuity, postoperative astigmatism, graft survival, and intraoperative and postoperative complications. CONCLUSION: Residents can be introduced to penetrating keratoplasty and achieve surgical success with intraoperative and postoperative complication rates similar to those previously published. PMID- 15364713 TI - The effect of nerve growth factor on corneal sensitivity after laser in situ keratomileusis. AB - OBJECTIVE: To determine if topically administered nerve growth factor (NGF) plays a role in accelerating the recovery of corneal sensitivity after laser in situ keratomileusis (LASIK). METHODS: A prospective double-masked study comparing the effect of topical NGF with balanced salt solution on corneal sensitivity after LASIK in rabbits. Preoperative and postoperative corneal sensitivities were assessed using an esthesiometer (Cochet-Bonnet esthesiometer). RESULTS: Eyes that were treated with topical NGF demonstrated an earlier and faster recovery of corneal sensitivity after LASIK (P =.007). A statistically significant difference in corneal sensitivity was found between the topical NGF and control group postoperatively at 2 (P =.01), 3 (P =.03), and 4 (P =.03) weeks. CONCLUSION: Topically administered NGF may play a significant role in accelerating corneal reinnervation after LASIK. Clinical Relevance Our results showed that topical NGF had beneficial effects in the early recovery of corneal sensitivity after LASIK. Nerve growth factor can be a new therapeutic approach for dry eye syndrome after LASIK. PMID- 15364714 TI - Additivity of bimatoprost or travoprost to latanoprost in glaucomatous monkey eyes. AB - OBJECTIVE: To compare the ocular hypotensive effect of the commercially available preparations of bimatoprost or travoprost added to latanoprost in monkey eyes with laser-induced unilateral glaucoma. METHODS: Four monkeys with unilateral laser-induced glaucoma were used in each treatment group and received drops in the glaucomatous eye only. Intraocular pressure (IOP) was measured hourly for 6 hours, beginning at 9:30 am on day 1 (untreated baseline), days 6 and 7 (single agent therapy), and days 13 and 14 (2-drug combination therapy). On days 2 through 7, 1 drop of the scheduled single agent was given immediately after the 9:30 am IOP measurement, and on days 8 through 14, the second scheduled drug was given 5 minutes after the first. The following 5 different dosing protocols were studied: latanoprost with bimatoprost added, bimatoprost with latanoprost added, latanoprost with travoprost added, travoprost with latanoprost added, and latanoprost with a second dose of latanoprost added. RESULTS: There were no statistically significant (P =.95) differences among the mean baseline IOPs in any of the 5 treatment groups. When applied as single agents, latanoprost, bimatoprost, and travoprost all produced significant (P<.05) and equivalent (P =.98) reductions in IOP. The mean +/-SEM maximum reduction (P<.05) from baseline IOP was 7.0 +/- 0.4 mm Hg (20% reduction) with travoprost alone, 6.5 +/- 1.6 mm Hg (18%) with bimatoprost alone, and 7.5 +/- 1.0 mm Hg (22%) with latanoprost alone. The mean +/-SEM maximum additive reductions in IOP were 3.0 +/- 0.6 mm Hg (P<.05) for travoprost added to latanoprost; 2.0 +/- 0.4 mm Hg (P<.05) for latanoprost added to travoprost; 4.8 +/- 1.3 mm Hg (P<.05) for bimatoprost added to latanoprost; 4.3 +/- 0.6 mm Hg (P<.05) for latanoprost added to bimatoprost; and 0.3 +/- 0.5 mm Hg (P>.60) for latanoprost added to itself. The combination of bimatoprost and latanoprost produced a greater (P<.05) lowering of IOP at trough and peak than the combination of travoprost and latanoprost. CONCLUSIONS: Latanoprost, bimatoprost, and travoprost used as monotherapy produced significant and equivalent reductions in IOP in glaucomatous monkey eyes. The IOP effects of the commercial concentrations of bimatoprost or travoprost were additive to that of latanoprost, with bimatoprost showing a greater additive response than travoprost. Clinical Relevance Because treatment with multiple medications is common among patients with glaucoma, determining which glaucoma medications produce an additive ocular hypotensive response when used in combination has practical implications for clinicians. PMID- 15364715 TI - Hypoxia-inducible factor 1alpha in the glaucomatous retina and optic nerve head. AB - OBJECTIVE: To examine tissue hypoxia in the retina and optic nerve head of glaucomatous eyes by the assessment of a transcription factor, hypoxia-inducible factor 1alpha (HIF-1alpha), which is tightly regulated by the cellular oxygen concentration. METHODS: Using immunohistochemical analysis, the cellular localization of HIF-1alpha was studied in the retina and optic nerve head of 28 human donor eyes with glaucoma compared with 20 control eyes from healthy donors matched for several characteristics. The relationship between the retinal regions that exhibited immunostaining for HIF-1alpha and functional damage was examined using visual field data. RESULTS: There was an increase in the immunostaining for HIF-1alpha in the retina and optic nerve head of glaucomatous donor eyes compared with the control eyes. In addition, the retinal location of the increased immunostaining for HIF-1alpha in some of the glaucomatous eyes was closely concordant with the location of visual field defects recorded in these eyes. CONCLUSIONS: Because the regions of HIF-1alpha induction represent the areas of decreased oxygen delivery and hypoxic stress, information obtained from this study provides direct evidence that tissue hypoxia is present in the retina and optic nerve head of glaucomatous eyes, and hypoxic signaling is a likely component of the pathogenic mechanisms of glaucomatous neurodegeneration. Clinical Relevance These findings support the presence of tissue hypoxia in the retina and optic nerve head of glaucomatous patients. PMID- 15364716 TI - Effectiveness of vitamin D analogues in treating large tumors and during prolonged use in murine retinoblastoma models. AB - OBJECTIVE: To investigate the effectiveness of the vitamin D analogues 1,25 (OH)(2)-16-ene-23-yne vitamin D(3) (16,23-D(3)) and 1alpha-hydroxyvitamin D(2) (1alpha-OH-D(2)) in inhibiting retinoblastoma growth in large tumors in a xenograft model and with prolonged use in a transgenic model. METHODS: For the large-tumor study, the xenograft athymic mouse/human retinoblastoma cell (Y-79) model was used. Subcutaneous tumors were allowed to grow to an average volume of 1600 mm(3). Systemic treatment with 1 of the vitamin D analogues or with vehicle (control groups) was carried out for 5 weeks. For the long-term study, transgenic beta-luteinizing hormone-large T antigen (LHbeta-Tag) mice were systemically treated with 1 of the 2 compounds or vehicle (control groups) for up to 15 weeks. Tumor size and signs of toxicity were assessed. RESULTS: In the large-tumor study, tumor volume ratios for the 1alpha-OH-D(2) and 16,23-D(3) groups were significantly lower than those for controls (P<.002). No significant differences in tumor volume were seen between the 1alpha-OH-D(2) and 16,23-D(3) groups (P =.15). In the long-term study, the 1alpha-OH-D(2) group showed significantly smaller tumor size compared with its control (P<.001). No significant difference was seen between the 16,23-D(3) group and its control. Some toxic effects related to hypercalcemia were seen in both studies. CONCLUSIONS: In athymic mice in the large-tumor study, both 1alpha-OH-D(2) and 16,23-D(3) were effective in inhibiting tumor growth compared with controls. In the long-term study, 1alpha-OH D(2) inhibited tumor growth but 16,23-D(3) did not. Effective doses of both compounds caused hypercalcemia and a significant increase in mortality. Clinical Relevance Use of 1alpha-OH-D(2) inhibited tumor growth in large tumors and with long-term treatment compared with controls. Because of hypercalcemia-related toxic effects seen in the present experiments, in clinical trials, serum calcium levels should be carefully monitored. This analogue may require use with drugs that lower serum calcium levels or use of relatively lower doses or skipped doses. The ideal alternative solution would be to identify vitamin D analogues that retain the antineoplastic action without the calcemic activity. PMID- 15364718 TI - Blinking Sam: the ocular afflictions of Dr. Samuel Johnson. AB - The poor health of Samuel Johnson (1709-1784) has fascinated the public for more than 200 years. The illnesses of few famous men, with the possible exception of Napoleon, have attracted more speculation. Johnson was an outstanding 18th century literary figure, an essayist, novelist, and poet, and is particularly famous as the creator of the first important dictionary of the English language. His writings and those of his physicians and friends, particularly his biographer, James Boswell, provide an intimate account of a cultural icon. PMID- 15364717 TI - Effectiveness of 1alpha-hydroxyvitamin D2 in inhibiting tumor growth in a murine transgenic pigmented ocular tumor model. AB - OBJECTIVE: To study the effectiveness of the vitamin D analogue 1alpha hydroxyvitamin D(2) (1alpha-OH-D(2)) in inhibiting ocular tumor growth in transgenic "Tyr-Tag" mice that developed pigmented ocular tumors produced with the simian virus 40 T and t antigens under the control of the mouse tyrosinase gene. These animals develop pigmented intraocular tumors primarily from the retinal pigment epithelium that closely resemble the histologic features and growth pattern of human choroidal melanoma. METHODS: A total of 73 Tyr-Tag transgenic mice between 6 and 7 weeks old were randomly assigned by sex and litter to 3 treatment groups to receive 0.05 microg/d, 0.1 microg/d, or 0.2 microg/d of 1alpha-OH-D(2); a control group received vehicle (coconut oil). The drug was administered by oral gavage 5 times a week for 5 weeks. The animals were then euthanized and their eyes were enucleated and processed histologically. Three serial sections from each eye were examined microscopically and the mean tumor area measured using Optimus software version 6.5 (Media Cybernetics LP, Silver Spring, Md). Toxic adverse effects were assessed on the basis of mortality, weight loss, and serum calcium levels. RESULTS: The mean tumor size in the 0.1- microg/d and 0.2- microg/d dose groups was smaller than in the controls (P<.001). No significant difference was seen between the 0.05- microg/d dose group and the control group (P =.64). Survival for the 0.1- microg/d and 0.2- microg/d dose groups was lower than for the controls (95% in the controls vs 85.7% and 73.7%, respectively; P<.01). CONCLUSION: In the Tyr-Tag transgenic mouse, 1alpha-OH-D(2) inhibits pigmented ocular tumor growth at moderate drug levels with relatively low mortality. Clinical Relevance Vitamin D analogues merit further preclinical study in the treatment of ocular melanoma. PMID- 15364719 TI - Should physicians prioritize their activities to address patients' wants or patients' needs? PMID- 15364720 TI - The role of tabloids in ophthalmic education: pro. PMID- 15364721 TI - The role of tabloids in ophthalmic education: con. PMID- 15364723 TI - Clinicopathologic correlation of successfully treated choroidal neovascularization lying within the notch of a large serous retinal pigment epithelial detachment. PMID- 15364724 TI - Allergic fungal sinusitis with unilateral eye involvement. PMID- 15364725 TI - Sequestration and late activation of lenticular Candida abscess in premature infants. PMID- 15364726 TI - Vitreous opacities and retinal vascular abnormalities in Gaucher disease. PMID- 15364727 TI - Choroidal neovascularization after central retinal vein occlusion in a 47-year old man. PMID- 15364728 TI - Neoplastic corneal pannus in sebaceous cell carcinoma. PMID- 15364729 TI - Gaze-evoked amaurosis associated with an intracranial aneurysm. PMID- 15364730 TI - The ocular complications of smallpox and smallpox immunization. PMID- 15364732 TI - Vitreous and aqueous penetration of orally administered gatifloxacin in humans. PMID- 15364734 TI - How (why) does oxygen work in advanced COPD? PMID- 15364735 TI - Is a charge a cost if nobody pays it? PMID- 15364736 TI - Core roll preparations and the pathologist as consultant. PMID- 15364737 TI - Complications of CT scan-guided lung biopsy: lesion size and depth matter. PMID- 15364738 TI - Which exercise test should be used for patients with symptomatic COPD? PMID- 15364739 TI - Treatment of severe acute respiratory syndrome. PMID- 15364740 TI - The evolving paradigm of hyperglycemia and critical illness. PMID- 15364741 TI - Role of interleukin-8 in the pathogenesis and treatment of COPD. PMID- 15364742 TI - Cardiac sympathetic nerve activity can detect congestive heart failure sensitively in patients with hypertrophic cardiomyopathy. AB - STUDY OBJECTIVES: Cardiac sympathetic nerve dysfunction is related to poor clinical outcome in patients with several different heart diseases. However, it is not clear whether cardiac sympathetic nerve activity is useful for predicting the onset of congestive heart failure (CHF) in patients with hypertrophic cardiomyopathy (HCM). The aim of this study was to examine the prognostic value of performing (123)I-labeled metaiodobenzylguanidine (MIBG) scintigraphy in patients with HCM in comparison with other conventional prognostic variables. METHODS: (123)I-labeled MIBG images were obtained from 84 HCM patients without prior CHF. After measurement of cardiac function, the patients were followed up for 9 to 86 months in our hospital. RESULTS: According to the cutoff values for the heart/mediastinum ratio (H/M) on delayed images of control subjects (ie, mean - 1 and 2 SDs), the patients were subdivided into the following three groups: group A (H/M, > 2.11; 34 patients); group B (H/M, < 1.86 to /= 75 years (OR, 5.68; 95% CI, 2.87 to 11.23; p < 0.0001) emerged as predictors of elevated levels of d-dimer, and only NVAF (OR, 10.30; 95% CI, 5.67 to 18.72; p < 0.0001) emerged as a predictor of elevated levels of beta-TG. CONCLUSIONS: NVAF patients whose conditions were complicated with risk factors for embolism could be in the prothrombotic state. Advanced age is a strong predictor of the prothrombotic state in NVAF patients. PMID- 15364744 TI - Hand-carried ultrasound improves the bedside cardiovascular examination. AB - OBJECTIVES: We assessed the clinical utility of hand-carried cardiac ultrasound (HCU) devices to assist physicians in the diagnosis of cardiovascular disease. MATERIALS AND METHODS: We reviewed 42 articles published from 1978 to 2004. RESULTS: The capability and simplicity of the HCU device assist physicians in the diagnosis of cardiovascular disease at the initial patients contact. HCU is particularly useful in the setting of emergency or critical care, community screening, or in remote areas with limited access to health care. CONCLUSION: The inherent limitations of the physical examination as well as the reduced focus and training in physical diagnosis of current and recent medical school graduates has set the stage for the HCU device to modify traditional medical practices by complementing the physical examination with real-time cardiovascular imaging. PMID- 15364745 TI - Intracoronary verapamil rapidly terminates reperfusion tachyarrhythmias in acute myocardial infarction. AB - BACKGROUND: The restoration of coronary flow after transient ischemia immediately induces life-threatening ventricular tachyarrhythmias. Although most of these arrhythmias disappear spontaneously, some of them induce serious hemodynamic changes. This retrospective study investigates the efficacy of therapy with intracoronary verapamil to terminate reperfusion-induced ventricular tachyarrhythmias in patients with acute myocardial infarction (AMI). METHODS AND RESULTS: Between February 1992 and February 2003, 390 patients with a diagnosis of AMI were enrolled into the study. All patients received mechanical revascularization therapy within 6 h of onset of symptoms, and 109 patients experienced reperfusion-induced tachyarrhythmias. A subset of these patients was treated with intracoronary verapamil (0.25 to 1.0 mg) to terminate the reperfusion-induced tachyarrhythmia. They were evaluated for immediate termination of the tachyarrhythmias, hemodynamic changes, resumption rates, and major complications. Thirty-one patients (28%) were treated with intracoronary verapamil for the immediate termination of reperfusion-induced ventricular tachyarrhythmias. These tachyarrhythmias included 6 premature ventricular contractions, 19 accelerated idioventricular rhythms, 3 ventricular tachycardias, 2 ventricular fibrillations (VFs), and 1 torsades de pointes. Intracoronary verapamil was effective in rapidly terminating all reperfusion-induced arrhythmias except for VFs. The side effects of treatment included temporary hypotension (two patients) and bradycardia (one patient), although all patients recovered spontaneously. No major complications were induced by the intracoronary use of verapamil, and no resumptions of arrhythmias were documented. CONCLUSION: Intracoronary administration of verapamil can safely terminate reperfusion induced ventricular tachyarrhythmias in a rapid manner. However, this effect was not seen for reperfusion-induced VF. PMID- 15364746 TI - Mitral valve repair in patients with low left ventricular ejection fractions: early and late results. AB - STUDY OBJECTIVES: This retrospective study was performed to examine the outcome of mitral valve repair (ie, mitral valvuloplasty [MVP]) in relation to preoperative low left ventricular ejection fraction (LVEF). DESIGN AND SETTINGS: From our series of 338 consecutive patients who underwent MVP between 1983 and 2001, we compared the course of 302 patients with preoperative LVEF of > 35% (group I) to that of 36 patients with LVEF of 500,000 patients undergo CABG each year. Numerous studies to date have suggested that postoperative atrial fibrillation occurs in 30 to 50% of patients, leading to significant morbidity, including hypotension, heart failure, thromboembolic complications, prolonged hospital stay, and increased hospital costs. The objective of this study was to assess the use of IV amiodarone in combination with oral amiodarone to reduce the incidence of postoperative atrial fibrillation. METHOD: From January 1999 to October 1999, 51 patients scheduled for CABG were randomly selected for participation in the amiodarone administration trial. IV amiodarone, 0.73 mg/min, was administered on call to the operating room for 48 h, followed by oral amiodarone, 400 mg q12h, for the next 3 days. The amiodarone group was case-control matched to the incidence of postoperative atrial fibrillation in 92 patients undergoing CABG using conventional medical therapy during the same period. The primary end point of this study was the incidence of postoperative atrial fibrillation, length of hospital stay, and hospital costs, compared to the control group undergoing CABG during the same time. RESULTS: Atrial fibrillation occurred in 3 of 51 patients (5.88%) in the amiodarone group, compared to 24 of 92 patients (26.08%) in the control group. Length of hospital stay in the amiodarone group was less than in the control group (5.3 days vs 6.7 days), with a trend toward decrease in hospital costs. CONCLUSION: The administration of IV amiodarone in conjunction with oral amiodarone for a total dose of 4,500 mg over 5 days appears to be a hemodynamically well-tolerated, safe, and effective treatment in decreasing the incidence of postoperative atrial fibrillation, shortening length of stay, and a trend toward lowering hospital costs, even in patients with significantly reduced left ventricular function (< 30%). A large multicenter study using IV and oral amiodarone should be pursued prior to deciding whether its use should become standard therapy in all patients undergoing CABG in order to decrease the incidence of postoperative atrial fibrillation. PMID- 15364748 TI - Long-term survival after videothoracoscopic lobectomy for stage I lung cancer. AB - STUDY OBJECTIVES: The aim of this study was to evaluate our personal experience and survival curves after video-assisted thoracic surgery (VATS) lobectomies for stage I lung cancer, in comparison with the results reported in existing literature. DESIGN: Retrospective analysis of our experience and an overview of literature. SETTING: Department of Surgery, San Giuseppe Hospital, University of Milan. PATIENTS: From October 1991 to December 2001, of 257 patients with clinical stage I lung cancer, 193 patients underwent VATS lobectomy, 7 patients were declared inoperable at thoracoscopic exploration, and 57 patients were converted (18 patients for oncologic reasons and 39 patients for technical reasons). RESULTS: We observed no intraoperative mortality and no recurrence. Survival data were analyzed with the Kaplan-Meier method along with a log-rank test for statistical significance. The global survival rates at 3 years and 5 years were 77.7% and 63.64%, respectively. T1N0 patients had a better survival curve at 3 years and 5 years (83.50% and 70.21%, respectively) compared to T2N0 patients (71.13% and 56.12%). Patients < 70 years of age had better 3-year and 5 year survival rates (82.37% and 73.32%, respectively) than those > 70 years of age (57.49% and 37.09%). This difference was statistically significant (p < 0.01). CONCLUSIONS: Our results after the VATS approach match the "best" results reported in literature following conventional surgery. Minimally invasive surgery seems to imply reduced tissue damage and decreased impairment of immunologic function. VATS lobectomy permits radical oncology as accurately as lobectomy by thoracotomy, provided that long-established standards are respected. In spite of technical difficulties, it could therefore become a valid alternative in treating stage I lung cancer. PMID- 15364749 TI - Surgical resection for lung cancer in the octogenarian. AB - BACKGROUND: As the US population ages, clinicians are increasingly confronted with octogenarians with resectable non-small cell lung cancer. Earlier reports documented substantial risk for surgical resection in this age group. METHODS: We reviewed our surgical experience in octogenarians who underwent curative resection from 1990 to 2003. RESULTS: Sixty-one patients underwent resection: 46 lobectomies, 6 segmentectomies, 5 wedge resections, and 4 pneumonectomies. There was one perioperative death (1.6%). The overall complication rate was 38% with a major complication rate of 13%. The average postoperative length of stay was 7 days. Overall 5-year survival was 38%, and 82% for stage IA patients. Patients with more advanced disease had a significantly worse survival. CONCLUSIONS: Appropriately selected octogenarians with early stage disease should be offered anatomic surgical resection for cure. These patients can anticipate a long-term survival, and should not be denied an operation on the basis of age alone. PMID- 15364750 TI - Usefulness of core roll preparations in immediate assessment of neoplastic lung lesions: comparison to conventional CT scan-guided lung fine-needle aspiration cytology. AB - INTRODUCTION: CT scan-guided fine-needle aspiration (FNA) of lung nodules is a well-established diagnostic technique. In our institution, both FNA and tissue core biopsy (using a 20-gauge needle) under CT scan guidance are routinely performed during the same procedure, and a preliminary diagnosis (an immediate assessment) is rendered. We compared core roll preparations (CRPs) with aspirate smears in the immediate assessment of pulmonary lesions and also assessed whether CRP resulted in the alteration of the histopathology of the core biopsy. STUDY DESIGN: Twenty-five cases of neoplastic pulmonary lesions diagnosed in CT scan guided lung FNA specimens, with core biopsies performed sequentially at the same visit for each patient, were evaluated. CRPs were made by lightly rolling the tissue core on a glass slide, followed by air-drying. Only stained slides (Diff Quik; Mercedes Medical; Sarasota, FL) were reviewed, and were scored for cellularity and morphology. RESULTS: CRPs in seven cases (28%) scored more diagnostic points than FNA smears, and were found to be better for cellularity and morphology compared to the corresponding FNA smears. The FNA smears scored more than CRPs in 10 cases (40%), while in 8 cases (32%) both CRPs and FNA smears scored equal diagnostic points. Using both CRP and FNA smears in the immediate assessment of lung biopsy specimens, we could assign a specific malignant histologic cell type in 23 of 25 cases (92%). In comparison, if the FNA smears were evaluated alone, we could assign a specific malignant histologic cell type in only 16 of 25 cases (64%). The CRP did not alter the histopathology of the core biopsy specimens in any of the above cases. CONCLUSION: The CRP complements the CT scan-guided lung FNA procedure in the immediate assessment of neoplastic lung lesions without altering the histopathology of core biopsy specimens. PMID- 15364751 TI - Percutaneous needle biopsy for small lung nodules beneath the rib under CT scan fluoroscopic guidance with gantry tilt. AB - OBJECTIVES: The present study was performed to evaluate the efficacy and safety of gantry tilting for the performance of lung biopsy of peripheral small lesions located beneath the rib. DESIGN: Interventional. MATERIALS AND METHODS: Our study was based on 22 of 237 lesions for which percutaneous needle biopsies of the lung were performed under CT scan-fluoroscopic guidance at our institution between January 2000 and August 2002. For these 22 lesions, a biopsy was performed with gantry tilt because a rib blocked the biopsy route even after trials to change the relationship between the target and the rib. The characteristics of each lesion, the success rate for obtaining an adequate specimen, and the ability to determine whether the lesion was malignant or benign were investigated, specific cell types were characterized, and the complications that were encountered were identified. RESULTS: In all 22 lesions, adequate specimens for cytopathologic evaluation were obtained using fine-needle aspiration biopsy, tissue core biopsies, or both. In 21 lesions, whether the lesion was malignant or benign was precisely diagnosed, and in 19 lesions the specific cell type was determined. No serious complications occurred. CONCLUSION: Percutaneous needle biopsy under CT scan-fluoroscopic guidance with gantry tilt is a useful and safe technique for the biopsy of small lung nodules located beneath the rib. PMID- 15364752 TI - Risk factors of pneumothorax and bleeding: multivariate analysis of 660 CT-guided coaxial cutting needle lung biopsies. AB - BACKGROUND: The results of studies identifying the risk factors for pneumothorax and bleeding in CT-guided coaxial lung needle biopsies were inconsistent and some were even contradictory. All reported series were small with patient populations averaging about 200. STUDY OBJECTIVES: To determine the risk factors for pneumothorax and bleeding after CT-guided coaxial cutting needle biopsy of lung lesions. DESIGN: Retrospective analysis. METHODS: We reviewed 660 biopsy procedures. The risk factors for pneumothorax and bleeding were determined by multivariate analysis of variables related to patient demographics, lung lesions, biopsy procedures, and the individual radiologist. RESULTS: The main complications were pneumothorax (23%; 155 of 660 procedures), chest tube insertion (1%; 9 of 660 procedures), and hemoptysis (4%; 26 of 660 procedures), with no patient mortality. The highest pneumothorax rate correlated with a lesion size of /= 2.1 cm, and the absence of pleural effusion. CONCLUSIONS: The risk factors for highest pneumothorax rate are lesion size /= 2.1 cm, and lung lesions not associated with a pleural effusion. PMID- 15364753 TI - Does whole-body 2-[18F]-fluoro-2-deoxy-D-glucose positron emission tomography have an advantage over thoracic positron emission tomography for staging patients with lung cancer? AB - BACKGROUND: Whole-body (WB) positron emission tomography (PET) with 2-[(18)F] fluoro-2-deoxy-D-glucose (FDG) is more accurate than other imaging studies for detecting lung cancer and extrathoracic metastatic disease. Thoracic PET (from the skull base through the kidneys) may be equally as useful as WB PET (skull base to mid-thigh). With the recent introduction of hybrid CT-PET systems, use of thoracic PET would minimize radiation dose. METHODS: A retrospective review of a series of WB PET scans performed in our department was performed to identify patients evaluated for a solitary pulmonary nodule or newly diagnosed lung cancer who had distant extracranial and extrathoracic metastases detected by PET. All patients with true extrathoracic metastases were documented by ancillary radiologic and clinical data. Patients were staged according to the American Joint Committee on Cancer TNM system based on findings within the confines of a thoracic PET and WB PET. Comparison was made between staging based on thoracic and WB PET to determine if there was a significant difference. RESULTS: Of 1,026 studies, distant extracranial metastases were described in 35 patients with lung cancer. Findings were determined to be false-positive in nine patients. Of the 26 patients with true metastases on WB PET, 25 patients had metastatic lesions within the confines of thoracic PET. Relative to WB PET, the sensitivity of thoracic PET is 96.2% (95% confidence interval, 1 to 99.3%) for detection of distant metastases. Only one patient had an isolated metastasis that was detected only by WB PET. This patient would have been staged IIIB by thoracic PET as opposed to stage IV by WB PET. CONCLUSIONS: Thoracic PET, when compared to WB PET, is 96.2% sensitive for detecting extrathoracic metastases in patients with newly diagnosed non-small cell lung cancer. PMID- 15364754 TI - The effect of tumor size on curability of stage I non-small cell lung cancers. AB - OBJECTIVE: The objective of this study was to determine the relationship between tumor size and curability of stage I non-small cell lung cancer. METHODS: From the Surveillance, Epidemiology, and End Results registry 2003, we identified all primary non-small cell lung cancer cases that were diagnosed prior to autopsy. Among these cases, we narrowed the focus to those diagnosed in 1988 or later, and to 7,620 patients who had undergone curative surgical resection. Kaplan-Meier survival curves were obtained for these stage I malignancies for five tumor size categories (ie, 5 to 15 mm, 16 to 25 mm, 26 to 35 mm, 36 to 45 mm, and > 45 mm). The 12-year Kaplan-Meier estimator of survival was used as a measure of lung cancer cure rate. RESULTS: Among 7,620 stage I cancers, cure rates decreased with increasing tumor size. The 12-year survival rates for patients with tumors 5 to 15 mm in diameter was 69% (95% confidence interval [CI], 64 to 74%), 63% for those with tumors 16 to 25 mm in diameter (95% CI, 60 to 67%), 58% for those with tumors 26 to 35 mm in diameter (95% CI, 54 to 61%), 53% for those with tumors 36 to 45 mm in diameter (95% CI, 48 to 57%), and 43% for those with tumors > 45 mm in diameter (95% CI, 39 to 48%). Cure rates were statistically significantly different for all tumor size categories (p < 0.05) except for the groups with tumors 26 to 35 mm and 36 to 45 mm in diameter (p = 0.10). CONCLUSIONS: Smaller tumor size at diagnosis is associated with improved curability within stage I non small cell lung cancers. These results suggest that further subclassification by size within stage I may be important. PMID- 15364755 TI - Physiologic responses to incremental and self-paced exercise in COPD: a comparison of three tests. AB - OBJECTIVES: To investigate cardiorespiratory and dyspnea responses to incremental and self-paced exercise tests in patients with COPD. DESIGN: A prospective within subject design was used. PATIENTS: Twenty stable subjects (15 men), with a mean (+/- SD) age of 64.0 +/- 7.5 years and moderate-to-severe COPD (ie, mean FEV(1), 0.8 +/- 0.3 L and 28.9 +/- 7.9% predicted) were studied. METHODS: Each subject completed a 6-min walk test (6MWT), an incremental shuttle walking test (ISWT), and a cycle ergometer test (CET), within a 2-week period. The tests were performed at least 24 h apart. Standardized encouragement was utilized in each test with the aim of maximizing performance. Heart rate (HR) and dyspnea were measured each minute throughout the tests, and pulse oximetric saturation (Spo(2)) was measured before and immediately after each test. The distances walked in the 6MWT and ISWT were compared to peak oxygen uptake (Vo(2)) values from the CET. RESULTS: HR increased linearly with increasing workload during the CET and ISWT, but increased alinearly with a disproportionate increase early in the 6MWT. In contrast, dyspnea scores increased linearly during the 6MWT, but increased alinearly with a disproportionate increase late during the CET and ISWT. Peak HR and dyspnea were not significantly different between tests. Spo(2) was significantly lower at the end of both walking tests compared to that at the end of the CET (p < 0.001). The distance walked in both the ISWT and 6MWT were related to peak Vo(2) values on the CET (for both tests, r = 0.73; p < 0.001). CONCLUSIONS: The patterns of response in HR and dyspnea seen during the 6MWT suggest that patients with COPD titrate exertion against dyspnea to achieve a peak tolerable intensity. This strategy is not possible in an externally paced ISWT or CET. However, it is a limited strategy, with performance converging at higher workloads. Similar peak exercise responses were achieved in the 6MWT, ISWT, and CET. Greater oxygen desaturation was observed during the field walking tests, suggesting that both the ISWT and 6MWT are more sensitive than the CET in detecting exercise-induced hypoxemia and in assessing ambulatory oxygen therapy needs. PMID- 15364756 TI - Effects of mechanical insufflation-exsufflation on respiratory parameters for patients with chronic airway secretion encumbrance. AB - STUDY OBJECTIVES: To analyze the physiologic effects and tolerance of mechanical insufflation-exsufflation (MI-E) for patients with chronic ventilatory failure of various etiologies. DESIGN: Prospective clinical trial. SETTING: Rehabilitation unit of a university hospital. PATIENTS OR PARTICIPANTS: Thirteen patients with amyotrophic lateral sclerosis (ALS), 9 patients with severe COPD, and 7 patients with other neuromuscular disorders (oNMDs) with chronic airway secretion encumbrance and decreases in oxyhemoglobin saturation (Spo(2)). INTERVENTIONS: Pressures of MI-E of 15 cm H(2)O, 30 cm H(2)O, and 40 cm H(2)O were cycled to each patient, with 3 s for insufflation and 4 s for exsufflation. One application was six cycles at each pressure for a total of three applications. MEASUREMENTS AND RESULTS: We continuously evaluated respiratory inductance plethysmography (RIP) and Spo(2) during every application. Peak cough flow (PCF) and dyspnea (Borg Scale) were also measured before the first and after the last application. The technique was well tolerated in all patient groups. Median Spo(2) improved significantly (p < 0.005) in all patient groups. Median PCF improved significantly (p < 0.005) in the ALS and oNMD groups from 170 to 200 L/min and from 180 to 220 L/min, respectively, and dyspnea improved significantly in the patients with oNMDs and patients with COPD from 3 to 1 and from 2 to 0.75, respectively. Breathing pattern characteristics (RIP) did not deteriorate after MI-E in any patient groups. Inspiratory flow limitation significantly decreased at the highest MI-E pressures for the ALS group. CONCLUSIONS: Our results confirm good tolerance and physiologic improvement in patients with restrictive disease and in patients with obstructive disease, suggesting that MI-E may be a potential complement to noninvasive ventilation for a wide variety of patient groups. PMID- 15364758 TI - Risk factors and natural history of habitual snoring. AB - STUDY OBJECTIVE: It has been suggested that habitual snoring (HS) has adverse health outcomes in children. We aimed to identify risk factors for HS and determine its natural history in primary school children. DESIGN: Cross sectional, population-based cohort study. SETTING: Twenty-seven primary schools located within the city limits of Hannover, Germany. PARTICIPANTS: Third-grade primary school children. MEASUREMENTS AND RESULTS: Snoring frequency and potential risk factors were investigated using parental questionnaires. Unadjusted and adjusted odds ratios (ORs) for HS and their 95% confidence intervals (CIs) were calculated. One year after the initial contact, snoring status was re-evaluated in habitual snorers. In total, 1,760 children were contacted, and 1,144 parents and their children (49% were girls) agreed to participate and returned a completed questionnaire. A body mass index >/= 90th percentile (OR, 3.5; 95% CI, 1.8 to 7.1), low maternal education (OR, 2.3; 95% CI, 1.1 to 4.7), regular daytime mouth breathing (OR, 7.4; 95% CI, 3.5 to 15.6), and a higher frequency of sore throats (OR, 17.6; 95% CI, 6.4 to 48.8) were independent risk factors for HS. Parental smoking and frequent infections were significantly but not independently associated with HS. The association of low maternal education and HS was higher in boys (OR, 4.4; 95% CI, 1.5 to 13.6; vs OR, 1.2; 95% CI, 0.4 to 3.6), while that of sore throats and HS was higher in girls (OR, 52.7; 95% CI, 6.0 to 460.2; vs OR, 13.3; 95% CI, 3.0 to 58.5). At follow-up, 39 of 80 eligible habitual snorers (48.8%) still snored regularly. Children who continued to snore differed significantly in maternal education, household smoking, snoring loudness, and prior ear, nose, throat surgery from those who had ceased to snore habitually. CONCLUSIONS: Socioeconomic status, obesity, signs of nasal obstruction, and pharyngeal problems were independent risk factors for HS in these primary school children. The expression of HS varied considerably over time. PMID- 15364759 TI - Baroreflex sensitivity in nonapneic snorers and control subjects before and after nasal continuous positive airway pressure. AB - HYPOTHESIS: We hypothesized that baroreflex sensitivity is decreased during wakefulness and non-rapid eye movement sleep in normotensive, nonapneic snorers who are otherwise healthy. Moreover, we hypothesized that nocturnal alterations in baroreflex sensitivity are abolished during the application of nasal continuous positive airway pressure (nCPAP). DESIGN: The sequencing technique was used to measure baroreflex sensitivity in 16 normotensive nonapneic snorers and 16 control subjects matched for age, height, weight, gender, and race. Subsequently, baroreflex sensitivity was measured in 12 of 16 snorers and 14 of 16 control subjects during the application of nCPAP. RESULTS: Mean (+/- SE) baroreflex sensitivity was reduced during sleep in the nonapneic snoring group (wakefulness, 20.99 +/- 1.46 ms/mm Hg; sleep, 15.85 +/- 1.49 ms/mm Hg), but not in the control group (wakefulness, 21.82 +/- 2.48 ms/mm Hg; sleep, 23.54 +/- 2.18 ms/mm Hg). This reduction was abolished by the application of nCPAP in the snoring group (before nCPAP therapy, 16.30 +/- 2.17 ms/mm Hg; during nCPAP therapy, 20.63 +/- 2.40 ms/mm Hg). The application of nCPAP did not alter baroreflex sensitivity in the control group (before nCPAP therapy, 23.54 +/- 2.18 ms/mm Hg; during nCPAP therapy, 22.56 +/- 1.73 ms/mm Hg). BP was not significantly different between the snoring and control groups either before or during nCPAP application. CONCLUSIONS: Our findings suggest that nocturnal alterations in baroreflex sensitivity may exist in nonapneic snoring subjects prior to alterations in other cardiovascular variables. PMID- 15364760 TI - Transition from epoprostenol and treprostinil to the oral endothelin receptor antagonist bosentan in patients with pulmonary hypertension. AB - STUDY OBJECTIVES: Prior to the availability of the oral endothelin antagonist bosentan, most patients with pulmonary arterial hypertension (PAH) were treated with continuously infused prostacyclins. Many patients receiving prostacyclins would have received bosentan if it had been available at the time of their diagnosis. Noninvasive criteria (symptoms, World Health Organization [WHO] functional class, 6-min walk test [6MWT] distances, and echocardiograms) are used to govern up-titration of prostacyclins and to assess response to bosentan. The purposes of this study were to see if some patients might be able to transition safely from prostacyclin to bosentan, and whether noninvasive criteria could be used to monitor this transition. METHODS: From January 2002 to July 2003, 23 stable patients with PAH attempted a transition from prostacyclin to bosentan over an 8-week period. 6MWT results, WHO class, and echocardiograms were recorded prior to transition and 1 month after successful transition. The transition was stopped and prostacyclin was resumed or up-titrated if any symptoms of PAH worsened. RESULTS: Of 23 candidates (19 female and 4 male; age range, 17 to 73 years), 15 patients were transitioned to bosentan. Of these patients, four patients experienced worsening symptoms (range, 7 weeks to 12 months after cessation of prostacyclin) and resumed treatment with prostacyclin. Of the remaining 11 patients, 2 patients had liver function abnormalities 3 months and 10 months after transition to bosentan, respectively; 9 patients remained on bosentan 3 to 16 months after prostacyclin cessation. Patients failing transition and resuming prostacyclin returned to their pretransition functional baseline. CONCLUSION: Nine of 23 carefully selected, stable patients with PAH receiving long-term prostacyclin were successfully transitioned to oral bosentan using noninvasive monitoring. No long-term adverse events were associated with failed transition attempts. Further studies need to be carried out to determine which patients are more likely to undergo the transition successfully. PMID- 15364761 TI - Reproducibility of cardiopulmonary exercise measurements in patients with pulmonary arterial hypertension. AB - BACKGROUND AND OBJECTIVES: As part of a recent study, cardiopulmonary exercise tests (CPETs) were used to evaluate and follow up patients with pulmonary arterial hypertension (PAH). These patients were more impaired than those in other published series evaluating CPET reproducibility. We used these patient tests to assess patient performance variability and evaluate reading variability. To achieve this end, six independent evaluators graded key CPET measurements in patients with PAH who underwent duplicate CPETs within 3 days of each other. SETTING AND PATIENTS: Over a 15-month period at two tertiary-care teaching hospitals, 42 patients with PAH underwent repeated, paired CPETs using cycle ergometry. INTERVENTIONS AND MEASUREMENTS: Each patient underwent one to six pairs of cycle ergometry tests to maximal tolerance. Each pair of tests was separated by 3 months, with each test in the pair separated by 1 to 3 days. Specific guidelines were given to the independent evaluators for the key measurements assessed from each CPET study: peak O(2) uptake (Vo(2)), peak heart rate, peak O(2) pulse, anaerobic threshold (AT), and end-tidal Po(2), end-tidal Pco(2), and the ventilatory equivalent for CO(2) at the AT (Ve/Vco(2)@AT). RESULTS: There were no fatalities or complications occurring among the 242 tests performed on 42 patients. The mean peak Vo(2) was 722 mL/min or 41% of predicted; 34 patients were Weber class C or D. Using the specific guidelines to measure the variability of measurements made by the six independent evaluators, the coefficients of variation were < 2.2% for peak Vo(2), peak heart rate, peak O(2) pulse, end-tidal values at the AT, and Ve/Vco(2)@AT, while for the AT, it was 8.5%. There were no significant differences in these measurements between the first and second tests of any pair or between the earlier and later sets of pairs. CONCLUSIONS: Using specific guidelines, key CPET measurements can be safely, reliably, and reproducibly assessed even in patients with severe exercise intolerance. PMID- 15364762 TI - Polymorphisms of renin-angiotensin system genes with high-altitude pulmonary edema in Japanese subjects. AB - STUDY OBJECTIVES: The renin-angiotensin system (RAS), including angiotensin converting enzyme (ACE) and angiotensin II type 1 receptor (AT(1)R), plays an important role in the pathogenesis of pulmonary hypertension, which is suggested to be critical in the development of high-altitude pulmonary edema (HAPE). Investigating the associations of the polymorphisms in the genes of RAS with HAPE is to elucidate the genetic background underlying this disease. DESIGN: A cross sectional, case-control study. SETTING: Shinshu University Hospital, Matsumoto, Japan. PARTICIPANTS: Forty-nine HAPE-susceptible (HAPE-s) subjects with a history of HAPE, and 55 healthy climbers with HAPE resistance (HAPE-r). INTERVENTIONS: Twenty-one of 49 HAPE-s subjects underwent right cardiac catheterization. MEASUREMENTS AND RESULTS: The insertion/deletion polymorphism in the ACE gene (ACE-I/D) was investigated by polymerase chain reaction (PCR). There was no significant difference of the distribution of the ACE-I/D polymorphism between the HAPE-s and HAPE-r groups. The A(1166)C and G(1517)T single-nucleotide polymorphisms (SNPs) in AT(1)R gene were investigated by the PCR following digested by corresponding restricted endonuclease enzymes. The distribution of the G(1517)T SNP was significantly different between the two groups (p = 0.012). The pulmonary hemodynamics of the 21 HAPE-s subjects were retrospectively examined. The pulmonary artery pressure (PAP), pulmonary vascular resistance (PVR), and PVR index (PVRI) were all significantly increased on hospital admission. Moreover, the PVR and PVRI were significantly higher in the HAPE-s subjects with D positivity than in the HAPE-s subjects with I positivity (PVR, p = 0.015; PVRI, p = 0.028), while the PAP did not show any significant difference between the two subgroups. CONCLUSIONS: The ACE-I/D polymorphism is not associated with HAPE susceptibility in Japanese subjects. The AT(1)R gene polymorphisms may likely associate with HAPE susceptibility. The D allele of the ACE-I/D polymorphism probably contributes to the hyperresponsive PVR and PVRI to acute hypoxia. PMID- 15364763 TI - Respiratory capacity course in patients with infantile spinal muscular atrophy. AB - STUDY OBJECTIVES: To describe the clinical and respiratory course in infantile spinal muscular atrophy (SMA) type I, type II, and type III, and to evaluate the respiratory needs for these patients, using noninvasive or tracheostomy ventilation. DESIGN: Retrospective cohort study. METHODS: We report 33 patients with SMA true type I (onset before age 3 months), 35 patients with SMA intermediate type I (onset between 3 months and 6 months), 100 patients with SMA type II (onset between 6 months and 18 months), 12 patients with SMA type III (onset after age 18 months). We report the clinical symptoms, respiratory course, and respiratory management: respiratory physiotherapy, periodic hyperinsufflation, nasal nocturnal ventilation (NNV), and tracheostomy. Also, we measured the FVC over several years during childhood and adolescence. RESULTS: In patients with SMA true type I, 82% of patients died, one third of whom underwent tracheostomy. In patients with SMA intermediate type I, 43% needed NNV, 57% underwent tracheostomy, and 26% died. In patients with SMA type II, 38% needed NNV, 15% underwent tracheostomy, and 4% died. In patients with SMA type III, respiratory impairment was moderate and began during the second decade of life. CONCLUSION: This data shows the progressively worsening course of restrictive respiratory insufficiency in patients with SMA, and the importance of early respiratory management to limit pulmonary complications and improve the quality of life for these patients. PMID- 15364764 TI - Inspiratory muscle weakness in diastolic dysfunction. AB - OBJECTIVES: To test the hypothesis that patients with well-documented diastolic dysfunction (DD) in the setting of normal systolic function will have inspiratory muscle weakness when compared to normal control subjects, and will experience dyspnea and tachypnea during exercise. BACKGROUND: Respiratory muscle weakness has been described in patients with (systolic) congestive heart failure; however, whether or not patients with DD may present with the findings of congestive heart failure is not known. METHODS: We selected for study 14 patients with DD previously referred for cardiopulmonary evaluation whose diagnosis had been confirmed by data obtained at cardiac catheterization. Seven control subjects matched for age, sex, and weight were recruited from the hospital community. Subjects performed both basic pulmonary function tests and tests of muscle strength: handgrip strength (Hgr), and maximal subatmospheric static inspiratory muscle pressure (Pimax). Subjects then performed a graded exercise test on a bicycle ergometer. Minute ventilation, oxygen consumption, carbon dioxide production, and heart rate were monitored continuously. Echocardiography was performed three times: before exercise, at a selected submaximal exercise level (20% of a predicted maximal workload), and at maximal exercise. Subjects rated their degree of dyspnea using the Borg scale at the same three time intervals. RESULTS: Pimax was - 102 +/- 17 cm H(2)O in control subjects, and - 77 +/- 19 cm H(2)O in patients with DD (p = 0.013) [mean +/- SD]. Hgr was similar between the groups. At the selected submaximal exercise level, patients with DD rated dyspnea to be 2.6 +/- 2.2 Borg scale units (control subjects, 0.5 +/- 0.8 Borg scale units). Hey plots described a rapid, shallow breathing pattern in patients with DD during exercise. Patients with DD and control subjects achieved similar maximal work loads. CONCLUSION: Patients with DD have diminished Pimax, adopt a rapid, shallow breathing pattern during exercise, and experience dyspnea at low work loads when compared to matched control subjects. PMID- 15364757 TI - Measurement of health-related quality of life in the national emphysema treatment trial. AB - PURPOSES: To evaluate two generic and two disease-specific measures of health related quality of life (QOL) using prerandomization data from the National Emphysema Treatment Trial (NETT). METHOD: The analyses used data collected from the 1,218 subjects who were randomized in the NETT. Patients completed evaluations before and after completion of the prerandomization phase of the NETT pulmonary rehabilitation program. Using data obtained prior to participation in the rehabilitation program, QOL measures were evaluated against physiologic and functional criteria using correlational analysis. The physiologic criteria included estimates of emphysema severity based on FEV(1) and measures of Pao(2) obtained with the subject at rest and breathing room air. Functional measures included the 6-min walk distance (6MWD), maximum work, and hospitalizations in the prior 3 months. RESULTS: Correlation coefficients between QOL measures ranged from -0.31 to 0.70. In comparison to normative samples, scores on general QOL measures were low, suggesting that the NETT participants were quite ill. All QOL measures were modestly but significantly correlated with FEV(1), maximum work, and 6MWD. Patients who had stayed overnight in a hospital in the prior 3 months reported lower QOL on average than those who had not been hospitalized. There were significant improvements for all QOL measures following the rehabilitation program, and improvements in QOL were correlated with improvements in 6MWD. COMMENT: The disease-specific and general QOL measures used in the NETT were correlated. Analyses suggested that these measures improved significantly following the rehabilitation phase of the NETT. PMID- 15364765 TI - Effectiveness of noninvasive positive pressure ventilation in the treatment of acute respiratory failure in severe acute respiratory syndrome. AB - OBJECTIVES: To study the effectiveness of noninvasive positive pressure ventilation (NIPPV) in the treatment of acute respiratory failure (ARF) in severe acute respiratory syndrome (SARS), and the associated infection risk. METHODS: All patients with the diagnosis of probable SARS admitted to a regional hospital in Hong Kong from March 9 to April 28, 2003, and who had SARS-related respiratory distress complications were recruited for NIPPV usage. The health status of all health-care workers working in the NIPPV wards was closely monitored, and consent was obtained to check serum for coronavirus serology. Patient outcomes and the risk of SARS transmission to health-care workers were assessed. RESULTS: NIPPV was applied to 20 patients (11 male patients) with ARF secondary to SARS. Mean age was 51.4 years, and mean acute physiology and chronic health evaluation II score was 5.35. Coronavirus serology was positive in 95% (19 of 20 patients). NIPPV was started 9.6 days (mean) from symptom onset, and mean duration of NIPPV usage was 84.3 h. Endotracheal intubation was avoided in 14 patients (70%), in whom the length of ICU stay was shorter (3.1 days vs 21.3 days, p < 0.001) and the chest radiography score within 24 h of NIPPV was lower (15.1 vs 22.5, p = 0.005) compared to intubated patients. Intubation avoidance was predicted by a marked reduction in respiratory rate (9.2 breaths/min) and supplemental oxygen requirement (3.1 L/min) within 24 h of NIPPV. Complications were few and reversible. There were no infections among the 105 health-care workers caring for the patients receiving NIPPV. CONCLUSIONS: NIPPV was effective in the treatment of ARF in the patients with SARS studied, and its use was safe for health-care workers. PMID- 15364766 TI - Assessment of physiologic variables and subjective comfort under different levels of pressure support ventilation. AB - STUDY OBJECTIVES: To evaluate the effects of 12 ventilator settings (pressure support ventilation [PSV] plus positive end-expiratory pressure [PEEP], 30 + 0 cm H(2)O; 25 + 5 cm H(2)O; 25 + 0 cm H(2)O; 20 + 5 cm H(2)O; 20 + 0 cm H(2)O; 15 + 5 cm H(2)O; 15 + 0 cm H(2)O; 10 + 5 cm H(2)O; 10 + 0 cm H(2)O; 5 + 5 cm H(2)O; 5 + 0 cm H(2)O; and 0 + 5 cm H(2)O) on physiologic variables; the percentage of ineffective efforts; patient comfort; and whether the diagnosis of COPD may influence results. DESIGN: Prospective, randomized, physiologic study. SETTING: Three weaning centers. PATIENTS: Thirty-six consecutive patients (20 patients with COPD). INTERVENTION: Patients were randomly submitted to the 12 settings. MEASUREMENTS AND RESULTS: Breathing pattern, respiratory drive (p0.1), arterial oxygen saturation (Sato(2)), heart rate, percentage of ineffective efforts per minute, patient comfort measured by means of a visual analogue scale (VAS), and BORG scale were recorded under each setting. Under different levels of assistance, breathing pattern, Sato(2), and p0.1 significantly and linearly changed (p < 0.0001) while VAS and BORG scale presented a significant (p = 0.027) U-shaped trend; high or low assistance caused the most discomfort. Under high levels of assistance, a higher (analysis of variance, p = 0.023) frequency of ineffective effort percentage was observed in the subgroup of 26 patients who presented this phenomenon. Breathing pattern significantly (p = 0.013) changed when compared to PSV alone (PSV plus zero end-expiratory pressure [ZEEP]) at the same total inspiratory pressure assistance (PSV plus PEEP). A huge variability among patients in breathing pattern and comfort was found under the setting rated as the most comfortable by patients. The diagnosis of COPD did not influence the overall results. CONCLUSIONS: The following conclusions are made: (1) physiologic variables followed a linear trend, while comfort followed a U-shaped trend under different levels of PSV (irrespective of COPD diagnosis); (2) high assistance caused an increase in ineffective efforts; (3) only the breathing pattern significantly changed when total assistance was given as PSV plus PEEP when compared to PSV alone (PSV plus ZEEP); and (4) the extreme levels of PSV are not associated with the best comfort. PMID- 15364767 TI - Experimental human endotoxemia is associated with depression of load-independent contractility indices: prevention by the lipid a analogue E5531. AB - OBJECTIVE: To evaluate the efficacy of a novel lipopolysaccharide (LPS) antagonist, E5531, in blocking LPS-induced cardiac responses including myocardial depression (as assessed by relatively load-independent echocardiographic indices of contractility) in a human model of experimental endotoxemia. DESIGN: Randomized, prospective, placebo-controlled, double-blind trial. SETTING: ICU procedure room. PARTICIPANTS: Thirty-two healthy, male volunteers. INTERVENTIONS: Administration of LPS (4 ng/kg) and either a placebo or one of four sequential doses of E5531 (100 microg, 250 microg, 500 microg, or 1,000 microg) followed by volumetric echocardiography before and during 4-L saline solution infusion (3 L over 3 h, followed by 1 L over 2 h). RESULTS: In addition to the generation of a hyperdynamic circulation throughout the study period, administration of LPS resulted in a biphasic contractility response. Ejection fraction (EF), rate corrected mean velocity of circumferential fiber shortening (Vcfc), peak systolic BP (SBP)/end-systolic volume index (ESVI) ratio, and end-systolic pressure (Pes)/ESVI ratio increased at the 3-h post-LPS assessment, compared to a control group of subjects receiving only similar amounts of saline solution (minimum p < 0.001). End-systolic myocardial wall stress (sigmaes)/ESVI ratio, one of the most load independent of the contractility indices, was unchanged. At 5 h after endotoxin, EF, Vcfc, SBP/ESVI, Pes/ESVI, and sigmaes/ESVI were all decreased (minimum p < 0.01), indicating myocardial depression. When present, early (3 h after LPS), apparent enhancement of myocardial contractility and later (5 h after LPS) myocardial depression were substantially blunted by administration of E5531 (minimum p < 0.025), typically in a concentration-dependent manner. CONCLUSIONS: Endotoxin generates significant myocardial depression when measured using highly load-independent indices of cardiac contractility. E5531 is a potent inhibitor of the early hyperdynamic cardiovascular and later myocardial depression response seen in experimental human endotoxemia. PMID- 15364768 TI - Evaluation of a modified percutaneous tracheostomy technique without bronchoscopic guidance. AB - BACKGROUND: Most complications of percutaneous tracheostomy are caused by failure to cannulate the trachea and injury to surrounding structures. Traditionally, the procedure has been performed under bronchoscopic assistance, which may interfere with the patient's ventilation and is cumbersome. A modification was described in which the subcutaneous tissue is bluntly dissected with a hemostat down to the pretracheal fascia. The procedure is then performed with the guidance of the operator's finger, making the routine use of a bronchoscope no longer necessary. METHODS: The modified technique was adopted and prospectively evaluated in an observational clinical study over a 30-month period, in patients requiring elective tracheostomy. Two commercially available kits were used. Patients' records were kept in files, and they were evaluated with regard to operative technique, complications, failure rate, and loss of airway. RESULTS: During the study period, 61 procedures were attempted. All were performed at the patients' bedside. In three patients (4.9%), the percutaneous procedure was deferred due to anatomic problems: cervical venous engorgement in one patient, and difficulty in dissection in another patient. In the third patient, the trachea could be felt, but the tube provided with the kit was not long enough. One patient had persistent wound bleeding, requiring revision in the operating room. No other procedure-related complications were reported. In three patients, early tube dislodgement occurred, but whether this was related to the percutaneous procedure is debatable. Bronchoscopy was not used. CONCLUSIONS: The modified percutaneous technique, with limited surgical dissection, without routine bronchoscopy, is simple and safe when performed by physicians with surgical training. It is relatively easy to learn, saves costs and operating room burden, and carries low morbidity rates. PMID- 15364769 TI - Gastric vs small-bowel feeding in critically ill children receiving mechanical ventilation: a randomized controlled trial. AB - STUDY OBJECTIVES: To determine the effect of feeding tube position (gastric vs small bowel) on adequacy of nutrient delivery and feeding complications, including microaspiration, in critically ill children. DESIGN: Randomized controlled trial. SETTING: Pediatric ICU in a university teaching hospital. PATIENTS: Seventy-four critically ill patients < 18 years of age receiving mechanical ventilation were randomized to receive gastric or small-bowel feeding. INTERVENTIONS: All feeding tubes were inserted at the bedside. Color, pH, and bilirubin concentration of the feeding tube aspirates were used to guide placement. Final tube position was confirmed radiographically. Continuous feedings were advanced to achieve a caloric goal based on age and body weight. Tracheal secretions were collected daily and tested for gastric pepsin by immunoassay. MEASUREMENTS AND RESULTS: Thirty-two patients were randomized to the gastric group, and 42 patients were randomized to the small-bowel group. Twelve patients exited the study because a small-bowel tube could not be placed at the bedside, leaving 30 patients in the small-bowel group. Gastric and small-bowel groups were similar at baseline in age, sex, percentage of ideal body weight, serum prealbumin concentration, and pediatric risk of mortality score. The percentage of daily caloric goal achieved was less in the gastric group compared to the small-bowel group (30 +/- 23% vs 47 +/- 22%, p < 0.01). No difference was found in the proportion of tracheal aspirates positive for pepsin between the gastric and small-bowel groups (50 of 146 aspirates vs 50 of 172 aspirates, respectively; p = 0.3). No differences were found in the frequency of feeding tube displacement, abdominal distension, vomiting, or diarrhea between groups. CONCLUSIONS: Small-bowel feeds allow a greater amount of nutrition to be successfully delivered to critically ill children. Small-bowel feeds do not prevent aspiration of gastric contents. PMID- 15364770 TI - Relationship of baseline glucose homeostasis to hyperglycemia during medical critical illness. AB - STUDY OBJECTIVE: To elucidate the relationship of baseline glucose control and acute stimuli with hyperglycemia during medical critical illness. DESIGN: Prospective cohort study. SETTING: Medical ICU (MICU) of a university affiliated hospital. PATIENTS: Convenience sample of 100 medical patients meeting criteria for severity of illness and anticipated length of stay and not admitted to the hospital for diabetic ketoacidosis or a hyperglycemic hyperosmolar state. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients were categorized as having normal, abnormal, or unevaluable baseline glucose control based on history and glycosylated hemoglobin (HbA1c). Data collection included blood glucose measurements within 120 h of MICU admission, and dosing of norepinephrine, corticosteroids, propofol, and carbohydrates. Average blood glucose and times over glycemic thresholds were calculated using linear interpolation. Hyperglycemia (glucose > 110 mg/dL) was pervasive in all groups. Among the 51 patients with normal baseline glucose control, HbA1c was correlated with hyperglycemic time (p < 0.01, R(2) = 0.15). Multiple regression found HbA1c, age, corticosteroid dose, and carbohydrate administration independently associated with hyperglycemic time (p < 0.05 for each, total R(2) = 0.49) in these patients, while body mass index, APACHE (acute physiology and chronic health evaluation) II, norepinephrine dose, propofol dose, gender, and sepsis were not associated with time > 110 mg/dL. Among normal subjects, HbA1c was independently predictive of peak and average glucose, and the fraction of time glucose was > 150 mg/dL and > 200 mg/dL (p < 0.05 for each). Patients with abnormal baseline glucose control had significantly more hyperglycemia than patients with normal baseline control. CONCLUSIONS: Even in patients without evidence of abnormal glucose homeostasis at baseline, hyperglycemia is common during critical illness. Time exposure to hyperglycemia is correlated with acute stressors and baseline glucose regulation, as characterized by HbA1c. Patients with low HbA1c levels are less disposed to hyperglycemia during severe illness than patients with higher, but still normal, HbA1c. PMID- 15364771 TI - Relation of epidermal growth factor receptor expression to mucus hypersecretion in diffuse panbronchiolitis. AB - STUDY OBJECT: Diffuse panbronchiolitis (DPB) is a hypersecretory airway disease, and the mechanism of mucus hypersecretion in DPB is poorly understood. Moreover, mucin synthesis in the airways has been reported to be regulated by neutrophilic inflammation-induced epidermal growth factor receptor (EGFR) expression, and the degranulation of goblet cells is known to be mediated by neutrophilic elastase. In this study, we examined the relationship between EGFR expression in the bronchiolar epithelium with neutrophilic inflammation and mucus hypersecretion in the tissues of DPB patients. DESIGN: The tissue specimens of 13 DPB patients and 6 healthy control subjects were examined by alcian blue/periodic acid-Schiff (AB/PAS) staining for mucous glycoconjugates, and by immunohistochemical staining for MUC5AC, EGFR, tumor necrosis factor-alpha, and CD16 on neutrophils. RESULTS: Neutrophilic inflammation was significantly higher in the tissue of DPB patients than in that of control subjects (p = 0.002). In the bronchiolar epithelium, goblet cell metaplasia, by AB/PAS staining and mucin MUC5AC expression, was significantly higher than that in control subjects (p = 0.001 and p = 0.002, respectively). In addition, the morphometric quantification of intraluminal mucus secretion showed that the areas of the bronchiolar lumen occupied by mucus secretion were significantly increased in the tissue of DPB patients (p = 0.001), suggesting goblet cell degranulation. EGFR expression was observed in the bronchiolar epithelium of DPB patients, but not in that of control subjects. CONCLUSIONS: In DPB, we suggest that mucus hypersecretion due to goblet cell metaplasia is closely associated with neutrophilic inflammation and the expression of EGFR. The study also shows that intraluminal secretion due to the degranulation of goblet cells degranulation is related to neutrophilic inflammation. PMID- 15364772 TI - Miniature intracardiac assist device provides more effective cardiac unloading and circulatory support during severe left heart failure than intraaortic balloon pumping. AB - BACKGROUND: Hemodynamic assistance with a miniature intracardiac pump may fill the treatment gap between use of an intraaortic balloon pump (IABP) and the current, more invasive ventricular assist devices. The objective of this study was to compare the hemodynamic efficacy of a miniature intracardiac pump device with that of IABP. METHODS AND RESULTS: Reversible acute mitral regurgitation (AMR) was induced in eight calves by stenting the mitral valve using a vena cava filter. Full and partial AMR assist were compared with maximum IABP support in each animal. In full-support mode, both assist systems increased cardiac output (miniature intracardiac pump, 13% [p < 0.05]; IABP, 3% [p < 0.05]), mean aortic pressure (miniature intracardiac pump, 13% [p < 0.05]; IABP, 8% [p < 0.05]), carotid artery flow (miniature intracardiac pump, 29% [p < 0.05]; IABP, 5% [difference not significant]), and coronary blood flow (miniature intracardiac pump, 25% [difference not significant]; IABP, 34% [p < 0.05]). Again in full support mode, both systems reduced left atrial pressure (miniature intracardiac pump, 2.4 mm Hg [p < 0.05]; IABP, 0.7 mm Hg [p < 0.05]), peak left ventricular (LV) pressure (miniature intracardiac pump, 13% [p < 0.05]; IABP, 5% [p < 0.05]), and external LV work (miniature intracardiac pump, 29% [p < 0.05]; IABP, 3% [p < 0.05]). Only full miniature intracardiac pump support reduced both end-diastolic LV volume (7%; p < 0.05) and end-systolic LV volume (10%; p < 0.05). IABP mainly improved coronary perfusion, while the miniature intracardiac pump proved more capable of genuinely unloading the LV. CONCLUSIONS: We conclude that during severe acute LV failure, the miniature intracardiac pump is capable of more effective cardiac unloading and circulatory support than IABP. PMID- 15364773 TI - Peripheral muscle strength training in COPD: a systematic review. AB - BACKGROUND: Skeletal muscle weakness, and the associated impact on exercise tolerance, provides a strong theoretical rationale for strength training intervention for people with COPD. AIM: The purpose of this review was to examine systematically the current evidence for peripheral muscle strength training in people with COPD. METHOD: Empirical trials and reviews relating to strength training in COPD were obtained by searching electronic databases and citation tracking, using the keywords of COPD with strength/resistance training and skeletal muscle. Two reviewers completed data extraction and quality assessment independently, using the PEDro (physiotherapy evidence database) scale and a checklist for review articles. Effect sizes and 95% confidence intervals were determined for empiric trials and metaanalysis used where appropriate. RESULTS: The search strategy yielded 13 articles (9 empirical trials and 4 reviews). Strength training was found to have strong evidence for improving upper body and leg strength. However, no strong evidence for strength training was found for other outcome measures. CONCLUSIONS: Further research is required to investigate the effects of strength training on functional activities, such as balance, upper limb function, self care, and participation in daily life. PMID- 15364774 TI - Long-term complications of congenital esophageal atresia and/or tracheoesophageal fistula. AB - Congenital esophageal atresia (EA) and/or tracheoesophageal fistula (TEF) are common congenital anomalies. Respiratory and GI complications occur frequently, and may persist lifelong. Late complications of EA/TEF include tracheomalacia, a recurrence of the TEF, esophageal stricture, and gastroesophageal reflux. These complications may lead to a brassy or honking-type cough, dysphagia, recurrent pneumonia, obstructive and restrictive ventilatory defects, and airway hyperreactivity. Aspiration should be excluded in children and adults with a history of EA/TEF who present with respiratory symptoms and/or recurrent lower respiratory infections, to prevent chronic pulmonary disease. PMID- 15364775 TI - Efficacy and safety of a monoclonal antibody recognizing interleukin-8 in COPD: a pilot study. AB - STUDY OBJECTIVE: To investigate the efficacy and safety of a fully human monoclonal antibody recognizing the chemokine interleukin (IL)-8 in patients with COPD. DESIGN: Randomized, double-blind, parallel-group, placebo-controlled trial. SETTING: Eighteen clinics/hospitals in the United States. PATIENTS: One hundred nine patients with stable COPD. INTERVENTIONS: Three IV infusions of either monoclonal antibody recognizing IL-8 (800-mg loading dose; 400-mg subsequent doses) or active buffer solution administered monthly over a 3-month period. MEASUREMENTS AND RESULTS: The differences in the transition dyspnea index (TDI) total score, the primary outcome measure, between fully human monoclonal IgG(2) antibody directed against IL-8 and placebo were 0.8, 1.0, 0.8, and 0.3 at week 2 (p = 0.046) and months 1 to 3, respectively. At all time points, the proportion of patients achieving >/= 1 point improvement in the TDI was greater for the monoclonal antibody group compared with the placebo group: 28% vs 11% at week 2 (p = 0.028). There were no significant differences observed for lung function, health status, 6-min walking distance, and adverse events between groups. CONCLUSIONS: The results of this phase 2 study suggest that neutralization of IL 8 with monoclonal antibody therapy may improve dyspnea in patients with COPD. These results support the further investigation of monoclonal antibody therapy targeting IL-8 for the treatment of this disease. PMID- 15364776 TI - Prehospital therapy with the platelet glycoprotein IIb/IIIa inhibitor eptifibatide in patients with suspected acute coronary syndromes: the Bochum feasibility study. AB - STUDY OBJECTIVES: To assess the practical application and safety of prehospital antithrombotic therapy with the glycoprotein (GP) IIb/IIIa inhibitor eptifibatide for patients with suspected acute coronary syndrome (ACS) or myocardial infarction (MI). DESIGN: Open-labeled pilot study. Patients with typical chest pain who were seen within 6 h of the onset of symptoms were enrolled in the mobile emergency ambulance. Patients were stratified by even/uneven days to receive standard treatment or standard treatment plus an IV bolus of eptifibatide (180 microg/kg body weight) followed by a continuous eptifibatide infusion (2 microg/kg/min). The main outcome measurement was a combination of prehospital or in-hospital death, reinfarction, revascularization of target vessels, and major bleeding complications. RESULTS: A total of 356 patients (age range, 29 to 75 years; women, 24.7%) were included in the analysis. On admission to the hospital, the diagnosis of ACS or MI was confirmed in approximately 60% of patients, and alternative diagnoses were made in 40% of patients. The rates of complications, including fatal and nonfatal complications occurring during transportation and during subsequent hospitalization, were similar in both study groups. The primary end point occurred in 11.8% of patients in the control group, and in 9.6% of those in the eptifibatide group (difference not significant). CONCLUSION: The prehospital administration of the GP IIb/IIIa inhibitor eptifibatide is feasible and safe in patients with clinically suspected ACS and MI. The benefit of this treatment has yet to be established in a large-scale multicenter study. PMID- 15364777 TI - Cardiopulmonary exercise testing and prognosis in chronic heart failure: a prognosticating algorithm for the individual patient. AB - The identification of individuals who are at high risk of chronic heart failure (HF) is a medical art of growing concern. Cardiopulmonary exercise stress testing (CPX) has become an important clinical tool to predict outcome. The value of peak oxygen consumption rests in the fact that it integrates elements of cardiac adaptations, and skeletal muscle, pulmonary, and endothelial dysfunctions more than other traditional prognostic indicators of chronic HF. Recently, exercise related ventilatory abnormalities have gained attention, stimulating scientific debate and an innovative perspective. This review, through a critical examination of previous experiences, will focus on the prognostic application of CPX, defining a proficient outline of treatment for the individual patient. PMID- 15364778 TI - Use of the Dumon Y-stent in the management of malignant disease involving the carina: a retrospective review of 86 patients. AB - OBJECTIVE: To study the efficacy of symptom relief and the safety of the Dumon Y stent for central airway obstruction in malignant main carinal involvement and in malignant tracheoesophageal fistulas. DESIGN: A retrospective review. SETTING: Thoracic endoscopy referral center. PATIENTS AND METHODS: The medical records of 86 patients who had undergone the placement of a Dumon Y-stent at our institution were reviewed and analyzed. RESULTS: Ninety Dumon Y-stents were placed in 86 patients to relieve symptoms of dyspnea, cough, and/or hemoptysis. Four of these patients required removal of that stent and replacement with a longer Dumon Y stent due to tumor progression. There were only two instances of procedure related adverse effects, which included coughing following stent placement lasting 1 week in one patient and stent migration in another patient. In the case of stent migration, the stent required immediate removal, and the patient experienced no adverse consequences. The remaining patients tolerated the stent well, and all experienced subjective symptomatic relief. There were no stent related deaths. The average duration of a stent after placement was 133 days. The median time of survival following stent insertion was 181 days. Forty-five percent of all patients died at 3 months, and 72% died at 6 months. CONCLUSION: The Dumon Y-stent proved to be useful and was well-tolerated in the management of malignant disease involving the main carina. It also provided successful palliation in patients with malignant tracheoesophageal fistulas. PMID- 15364779 TI - Endobronchial ultrasonography using a guide sheath increases the ability to diagnose peripheral pulmonary lesions endoscopically. AB - STUDY OBJECTIVE: To assess the ability of endobronchial ultrasonography (EBUS) using a guide sheath (EBUS-GS) to diagnose peripheral pulmonary lesions. METHOD: We devised a technique for EBUS-GS covering a miniature probe, and 150 lesions were evaluated in a prospective open study. In this procedure, the probe covered by a guide sheath is introduced into the lesion via the working channel of a bronchoscope. The probe is withdrawn, while the guide sheath is left in situ. A brush or biopsy forceps is introduced through the guide sheath into the lesion. RESULTS: One hundred sixteen of 150 EBUS-GS procedures (77%) were diagnostic. Cases in which the probe was located within the lesion had a significantly higher diagnostic yield (105 of 121 cases, 87%) than when the probe was located adjacent to it (8 of 19 cases, 42%) [p < 0.0001, chi(2)]. The diagnostic yield from EBUS GS in lesions 10 to 15 to 20 to 20 years, onset > 30 years ago, and time since last exposure > 7 years. More AB-positive patients reported respiratory symptoms (70% vs 26%, p < 0.05). High-resolution CT scans of AB-positive patients revealed a higher prevalence of parenchymal disease (70% vs 26%, p < 0.05). AB positive subjects had reduced pulmonary function compared to AB-negative subjects: FVC (86% vs 97% predicted), FEV(1) (77% vs 92% predicted, p < 0.05), and diffusion capacity of the lung for carbon monoxide (76% vs 104% predicted, p < 0.01). CONCLUSION: In individuals with a history of asbestos exposure, the presence of AB in BAL cells is associated with higher prevalence of parenchymal abnormalities, respiratory symptoms, and reduced pulmonary function. PMID- 15364781 TI - What does it mean when a patient says, "my asthma medication is not working?". PMID- 15364782 TI - A patient with new-onset seizure and mediastinal adenopathy. PMID- 15364783 TI - Bilateral pleural effusions and right pneumothorax in a 25-year-old man. PMID- 15364784 TI - Eosinophilic pneumonia as an initial manifestation of rheumatoid arthritis. AB - Rheumatoid arthritis is a systemic disease that may have pulmonary manifestations. We describe a case of eosinophilic pneumonia as the primary presentation of rheumatoid arthritis. While several cases of acute and chronic eosinophilic pneumonia have been reported in patients with preexisting rheumatoid arthritis, this is the first case reported in which the eosinophilic lung disease was the initial manifestation of systemic rheumatoid arthritis. PMID- 15364785 TI - Pregnancy and sarcoidosis: an insight into the pathogenesis of hypercalciuria. AB - Hypercalciuria with or without hypercalcemia is a well-known complication of sarcoidosis, the pathogenesis of which is not fully understood. Pregnancy is associated with physiologic alterations in calcium metabolism. These changes can further alter the derangement of calcium metabolism that occurs in sarcoidosis, if the two conditions coexist. We had the opportunity to study prospectively the changes in serum and urine calcium along with all the hormonal changes that occur during pregnancy in a young woman with sarcoidosis, who had hypercalciuria at presentation. We believe that an increased level of calcitriol is central to the calcium abnormalities in our patient. In her case, the increased calcitriol is derived from sarcoid granulomas and renal sources enhanced by the effect of estradiol and prolactin on the conversion of 25(OH)D to 1,25(OH)(2) D. She acquired hypoparathyroidism, with normal serum calcium, which probably was due to the direct suppression of parathyroid hormone (PTH) secretion by calcitriol. Finally, hypercalciuria is the result of the combined effect of hyperabsorption of calcium from the gut (the result of increased calcitriol levels leading to increased filtration of calcium) and decreased tubular reabsorption of calcium, as a result of undetectable PTH. PMID- 15364786 TI - Changing patterns in asbestos-induced lung disease. PMID- 15364787 TI - Failure to prove asbestos exposure produces obstructive lung disease. PMID- 15364788 TI - What is a MOSAIC study? PMID- 15364789 TI - The importance of bronchoscope reprocessing guidelines: raising the standard of care. PMID- 15364790 TI - Help for interpretation of cardiopulmonary exercise testing. PMID- 15364791 TI - Open-lung biopsy for ARDS patients. PMID- 15364792 TI - Racial differences in allergen sensitivity. PMID- 15364793 TI - Conventional vs endobronchial ultrasound-guided transbronchial needle aspiration of the mediastinum. PMID- 15364794 TI - Measuring the work of exercise. PMID- 15364795 TI - 25-year study of lung fibrosis following carmustine therapy for brain tumor in childhood. PMID- 15364796 TI - Effect of pravastatin on cardiovascular events in people with chronic kidney disease. AB - BACKGROUND: Limited data describe the cardiovascular benefit of HMG-CoA reductase inhibitors (statins) in people with moderate chronic kidney disease (CKD). The objective of this analysis was to determine whether pravastatin reduced the incidence of cardiovascular events in people with or at high risk for coronary disease and with concomitant moderate CKD. METHODS AND RESULTS: We analyzed data from the Pravastatin Pooling Project (PPP), a subject-level database combining results from 3 randomized trials of pravastatin (40 mg daily) versus placebo. Of 19 700 subjects, 4491 (22.8%) had moderate CKD, defined by an estimated glomerular filtration rate of 30 to 59.99 mL/min per 1.73 m2 body surface area. The primary outcome was time to myocardial infarction, coronary death, or percutaneous/surgical coronary revascularization. Moderate CKD was independently associated with an increased risk of the primary outcome (adjusted HR 1.26, 95% CI 1.07 to 1.49) compared with those with normal renal function. Among the 4491 subjects with moderate CKD, pravastatin significantly reduced the incidence of the primary outcome (HR 0.77, 95% CI 0.68 to 0.86), similar to the effect of pravastatin on the primary outcome in subjects with normal kidney function (HR 0.78, 95% CI 0.65 to 0.94). Pravastatin also appeared to reduce the total mortality rate in those with moderate CKD (adjusted HR 0.86, 95% CI 0.74 to 1.00, P=0.045). CONCLUSIONS: Pravastatin reduces cardiovascular event rates in people with or at risk for coronary disease and concomitant moderate CKD, many of whom have serum creatinine levels within the normal range. Given the high risk associated with CKD, the absolute benefit that resulted from use of pravastatin was greater than in those with normal renal function. PMID- 15364797 TI - Endothelial dysfunction and damage in congestive heart failure: relation of flow mediated dilation to circulating endothelial cells, plasma indexes of endothelial damage, and brain natriuretic peptide. AB - BACKGROUND: Congestive heart failure (CHF) is associated with endothelial perturbation (as defined by flow-mediated endothelial-dependent vasodilation [FMD, an index of endothelial dysfunction], circulating endothelial cells [CECs, an index of endothelial damage], or plasma indexes of endothelial damage/dysfunction [eg, von Willebrand factor (vWf) and soluble thrombomodulin (sTM)]) and raised plasma levels of brain natriuretic peptide (BNP, a peptide hormone associated with left ventricular systolic dysfunction and prognosis). However, the relations between these parameters are unclear. METHODS AND RESULTS: To test the hypothesis that there is a relation between endothelial perturbation (defined by FMD, CECs, vWf, and sTM) and BNP in CHF, we studied these indexes in 30 patients with CHF who were compared with 20 age-matched control subjects. FMD, CECs, plasma vWf, and BNP levels (but not sTM) were all abnormal in patients with CHF. There were significant inverse correlations between FMD and vWf (P=0.001), CECs (P=0.002) and BNP (P=0.006) as well as a positive correlation between CECs and vWf (P=0.032). In multivariate analysis, BNP (P<0.001) and FMD (P<0.001) were both independently associated with CHF. CONCLUSIONS: Ample evidence of endothelial cell damage/dysfunction in CHF cannot be fully explained by the variance in plasma BNP per se. Therefore, the routes by which these indexes influence the pathophysiology of CHF as well as predict adverse outcomes may be independent. PMID- 15364798 TI - Possible association of heart failure status with synthetic balance between aldosterone and dehydroepiandrosterone in human heart. AB - BACKGROUND: Aldosterone is produced not only in the adrenal gland but also in the extra-adrenal tissues, including failing human heart. This study examined the production of dehydroepiandrosterone (DHEA) in human heart and elucidated the possible physiological significance. Method and Results- Using left ventricular tissues obtained at autopsy, reverse transcription-polymerase chain reaction followed by Southern blot analysis revealed the gene expressions of CYP17. By measuring plasma aldosterone and DHEA levels at the coronary sinuses and aortic roots during cardiac catheterization, we found that DHEA but not aldosterone was secreted from control subjects (P<0.0001 and P=0.74, respectively), whereas aldosterone but not DHEA was secreted from patients with heart failure (P=0.0017 and P=0.67, respectively). To examine the significance of DHEA, we measured myocyte cell sizes and the gene expression of B-type natriuretic peptide (BNP), using a neonatal rat cardiocyte culture system. We found that DHEA (10(-8) mol/L) significantly inhibited the increase in myocyte cell sizes and BNP mRNA levels upregulated by endothelin-1 (P=0.031 and P<0.0001, respectively). CONCLUSIONS: CYP17 gene expression and production of DHEA were demonstrated in human control heart. Also, we found that cardiac production of DHEA was suppressed in failing heart. We postulated that DHEA and/or its metabolites exert a cardioprotective action through antihypertrophic effects. PMID- 15364799 TI - Suppression of acute and chronic rejection by hepatocyte growth factor in a murine model of cardiac transplantation: induction of tolerance and prevention of cardiac allograft vasculopathy. AB - BACKGROUND: Although treatment with immunosuppressive agents has contributed to overcoming acute rejection and improving the midterm survival of transplanted hearts, cardiac allograft vasculopathy (CAV) has remained the main cause of primary graft failure. Recent approaches have shown that hepatocyte growth factor (HGF) exhibits cardiotrophic functions. We therefore addressed whether HGF would regulate acute and chronic rejection in cardiac transplantation. METHODS AND RESULTS: We used a murine heterotopic cardiac transplantation model between fully incompatible strains and administered 500 microg x kg(-1) x d(-1) HGF during the initial 14 days after transplantation. The HGF-treated allografts showed significantly prolonged survival (42.3+/-4.1 days, P<0.001) compared with the controls (11.1+/-0.6 days), with tolerance induction in 47.4%. Histopathologically, the number of infiltrating cells was significantly decreased and myocardial necrosis was less prominent with a reduction of apoptosis in the allografts by HGF treatment during acute rejection. In the long-term surviving allografts, HGF significantly inhibited the development of CAV and interstitial fibrosis. With respect to intragraft cytokine mRNA expression, HGF treatment reduced the early expression of interferon-gamma and enhanced the expression of transforming growth factor-beta1 during the acute phase and of interleukin-10 continuously through the acute phase to the chronic phase. CONCLUSIONS: Our findings demonstrate that HGF can prolong the survival of allografts by its cardioprotective and immunomodulative potencies. Thus, HGF administration may constitute a new therapeutic approach to preventing cardiac graft failure that has not been overcome by conventional immunosuppressive agents. PMID- 15364800 TI - Different quantitative apoptotic traits in coronary atherosclerotic plaques from patients with stable angina pectoris and acute coronary syndromes. AB - BACKGROUND: Apoptosis in human atherosclerotic coronary plaques possibly causes plaque destabilization by contributing to the weakening and breaking down of the fibrous cap. We tested the hypothesis that apoptosis is quantitatively increased in unstable atherosclerotic plaques. METHODS AND RESULTS: We analyzed the expression of apoptotic genes such as BAX, CASP1, FAS, FAS L, FOS, MDM2, NFkB2, P53, PCNA, TERT, and XRCC1 in coronary plaques collected with directional coronary atherectomy from 15 patients with stable angina and 15 with acute coronary syndromes without ST elevation (ACS). Total RNA was extracted and cDNA was amplified with a specific set of primers and TaqMan probes. Apoptosis was also revealed by DNA laddering. To clarify the source of mRNAs, we performed in situ reverse transcriptase-polymerase chain reaction coupled with immunocytochemistry and found a substantial overlap between the mRNAs of the above genes and vascular smooth muscle cells. Gene expression analysis showed that the proapoptotic genes (ie, BAX, CASP1, FAS, FAS L, FOS, NFkB2, P53, PCNA) were significantly more expressed (P<0.001) in ACS plaques, whereas the antiapoptotic genes (ie, MDM2, TERT, XRCC1) were more transcribed (P<0.001) in stable angina plaques. Total gDNA gel electrophoresis identified a laddering pattern in the ACS plaques as evidence of end-point apoptosis. Western blotting substantially confirmed the above data. CONCLUSIONS: Our findings support the idea that ACS plaques are committed to apoptosis through an established meshwork of gene activation and inactivation, whereas stable angina plaques retain active cell homeostasis and repair mechanisms. PMID- 15364801 TI - Effect of aging and physical activity on left ventricular compliance. AB - BACKGROUND: Left ventricular compliance appears to decrease with aging, which may contribute to the high incidence of heart failure in the elderly. However, whether this change is an inevitable consequence of senescence or rather secondary to reduced physical activity is unknown. METHODS AND RESULTS: Twelve healthy sedentary seniors (69.8+/-3 years old; 6 women, 6 men) and 12 Masters athletes (67.8+/-3 years old; 6 women, 6 men) underwent pulmonary artery catheterization to define Starling and left ventricular pressure-volume curves. Data were compared with those obtained in 14 young but sedentary control subjects (28.9+/-5 years old; 7 women, 7 men). Pulmonary capillary wedge pressures and left ventricular end-diastolic volumes by use of echocardiography were measured at baseline, during decreased cardiac filling by use of lower-body negative pressure (-15 and -30 mm Hg), and after saline infusion (15 and 30 mL/kg). Stroke volume for any given filling pressure was greater in Masters athletes compared with the age-matched sedentary subjects, whereas contractility, as assessed by preload recruitable stroke work, was similar. There was substantially decreased left ventricular compliance in healthy but sedentary seniors compared with the young control subjects, which resulted in higher cardiac pressures for a given filling volume and higher myocardial wall stress for a given strain. The pressure volume curve for the Masters athletes was indistinguishable from that of the young, sedentary control subjects. CONCLUSIONS: A sedentary lifestyle during healthy aging is associated with decreased left ventricular compliance, leading to diminished diastolic performance. Prolonged, sustained endurance training preserves ventricular compliance with aging and may help to prevent heart failure in the elderly. PMID- 15364802 TI - Does the functional efficacy of skeletal myoblast transplantation extend to nonischemic cardiomyopathy? AB - BACKGROUND: The benefits of skeletal myoblast (SM) transplantation on infarcted myocardium have been investigated extensively; however, little is known about its effects in nonischemic cardiomyopathy models. To address this issue, we tested SM transplantation in CHF147 Syrian hamsters, a strain characterized by a delta sarcoglycan deficiency that phenotypically features the human setting of primary dilated cardiomyopathy. METHODS AND RESULTS: Cell culture techniques were used to prepare approximately 5x10(6) muscle cells from autologous tibialis anterior muscle, of which 50% were SMs (desmin staining). The cells were injected in 6 sites across the left ventricular wall (n=14). Control animals (n=12) received equivalent volumes of culture medium. Left ventricular systolic function was assessed in a blinded fashion from 2D echocardiographic left ventricular fractional area change, before transplantation, and 4 weeks later. Explanted hearts were processed for the detection of myotubes and quantification of fibrosis. Baseline functional data did not differ between the 2 groups. Four weeks after transplantation, 6 of the 10 surviving grafted hamsters were improved compared with 0 of the 8 survivors of the control group. This translated into a 6% decrease in fractional area change in controls compared with a 24% increase in cell-transplanted hamsters (P=0.001). Engrafted myotubes were consistently detected in all SM transplanted hearts by immunohistochemistry, whereas fibrosis was not worsened by cell injections. CONCLUSIONS: These data suggest that the functional benefits of SM transplantation might extend to nonischemic cardiomyopathy. PMID- 15364803 TI - Single-arm study of bridging therapy with low-molecular-weight heparin for patients at risk of arterial embolism who require temporary interruption of warfarin. AB - BACKGROUND: When warfarin is interrupted for surgery, low-molecular-weight heparin is often used as bridging therapy. However, this practice has never been evaluated in a large prospective study. This study was designed to assess the efficacy and safety of bridging therapy with low-molecular-weight heparin initiated out of hospital. METHODS AND RESULTS: This was a prospective, multicenter, single-arm cohort study of patients at high risk of arterial embolism (prosthetic valves and atrial fibrillation with a major risk factor). Warfarin was held for 5 days preoperatively. Low-molecular-weight heparin was given 3 days preoperatively and at least 4 days postoperatively. Patients were followed up for 3 months for thromboembolism and bleeding. Eleven Canadian tertiary care academic centers participated; 224 patients were enrolled. Eight patients (3.6%; 95% CI, 1.8 to 6.9) had an episode of thromboembolism, of which 2 (0.9%; 95% CI, 0.2 to 3.2) were judged to be due to cardioembolism. Of these 8 episodes of thromboembolism, 6 occurred in patients who had warfarin deferred or withdrawn because of bleeding. There were 15 episodes of major bleeding (6.7%; 95% CI, 4.1 to 10.8): 8 occurred intraoperatively or early postoperatively before low-molecular-weight heparin was restarted, 5 occurred in the first postoperative week after low-molecular-weight heparin was restarted, and 2 occurred well after low-molecular-weight heparin was stopped. There were no deaths. CONCLUSIONS: Bridging therapy with subcutaneous low-molecular-weight heparin is feasible; however, the optimal approach for the management of patients who require temporary interruption of warfarin to have invasive procedures is uncertain. PMID- 15364804 TI - Cardiac specific increase in aldosterone production induces coronary dysfunction in aldosterone synthase-transgenic mice. AB - BACKGROUND: Elevated circulating aldosterone level is associated with impaired cardiovascular function. Although the mechanisms are not fully understood, aldosterone antagonists decrease total and cardiovascular mortality in heart failure and myocardial infarction. Aldosterone induces cardiac fibrosis in experimental models, and it is synthesized locally in rat heart. These observations suggest pathological effects of aldosterone in heart that remain unclear. METHODS AND RESULTS: Transgenic mice (TG) that overexpress the terminal enzyme of aldosterone biosynthesis, aldosterone synthase (AS), in heart have been raised by gene targeting with the alpha-myosin heavy chain promoter. AS mRNA increased 100-fold and aldosterone concentration 1.7-fold in hearts of male TG mice relative to wild-type. No structural or myocardial alterations were evidenced, because ventricle/body weight, AT1 and AT2 receptor binding, and collagen content were unchanged in TG. No alteration in cardiac function was evidenced by echocardiography, isolated perfused heart, or whole-cell patch clamp experiments. In contrast, coronary function was impaired, because basal coronary flow was decreased in isolated perfused heart (-55% of baseline values), and vasodilatation to acetylcholine, bradykinin, and sodium nitroprusside was decreased by 75%, 60%, and 75%, respectively, in TG mice compared with wild-type, showing that the defect was not related to NO production. CONCLUSIONS: Increased cardiac aldosterone production in male mice induces a major coronary endothelium independent dysfunction with no detectable alterations in cardiac structure and function. However, coronary dysfunction may be harmful for coronary adaptation to increased flow demand. PMID- 15364805 TI - Red wine polyphenolic compounds strongly inhibit pro-matrix metalloproteinase-2 expression and its activation in response to thrombin via direct inhibition of membrane type 1-matrix metalloproteinase in vascular smooth muscle cells. AB - BACKGROUND: Regular consumption of moderate amounts of red wine is associated with a reduced risk of coronary disease. Matrix metalloproteinases (MMPs) that participate in extracellular matrix degradation have been involved in atherosclerotic plaque growth and instability. The present study examined whether red wine polyphenolic compounds (RWPCs) inhibit activation of MMP-2, a major gelatinase, in vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: Expression of pro-MMP-2 was assessed by Western and Northern blot analyses; MMP-2 activity was assessed by zymography and cell invasion by a modified Boyden's chamber assay. High levels of pro-MMP-2 and low levels of MMP-2 activity were found in conditioned medium from unstimulated VSMCs. Thrombin induced cell associated pro-MMP-2 protein expression and MMP-2 activity in conditioned medium of VSMCs. The stimulatory effect of thrombin on MMP-2 activation was prevented by RWPCs in a concentration-dependent and reversible manner. Thrombin markedly increased cell-associated membrane type 1 (MT1)-MMP activity, the physiological activator of pro-MMP-2, and this response was not affected by RWPCs. However, addition of RWPCs directly to MT1-MMP abolished its metalloproteinase activity in a reversible manner. Finally, matrix invasion of VSMCs was stimulated by thrombin, and this response was prevented by RWPCs as efficiently as a broad spectrum MMP inhibitor. CONCLUSIONS: The present findings demonstrate that RWPCs effectively inhibit thrombin-induced matrix invasion of VSMCs, most likely by preventing the expression and activation of MMP-2 via direct inhibition of MT1 MMP activity. The inhibitory effect of RWPCs on the activation of pro-MMP-2 and matrix degradation might contribute to their beneficial effects on the cardiovascular system. PMID- 15364806 TI - Probucol protects against smooth muscle cell proliferation by upregulating heme oxygenase-1. AB - BACKGROUND: Evidence suggests that induction of heme oxygenase-1 (HO-1) inhibits proliferation of vascular smooth muscle cells and intimal thickening after arterial injury, and therapeutic molecules induce HO-1. Probucol is the only oral drug that inhibits restenosis in humans and intimal thickening in animals, although its underlying mechanism remains unclear. METHODS AND RESULTS: Aortas were harvested from New Zealand White rabbits fed normal or 0.75% (wt/wt) probucol-fortified chow, with or without endothelial denudation of the abdominal aorta on day 21, and analyzed for heme oxygenase and apoptosis. Uninjured aortas were harvested on day 21 and balloon-injured aortas on days 22 and 25. Probucol significantly increased mRNA of HO-1 assessed by real-time PCR and HO activity in aortas at all time points. Probucol also enhanced apoptosis of medial cells in the injured aorta, as evidenced by the TUNEL assay. Furthermore, probucol (100 micromol/L) increased HO-1 mRNA and HO activity when added to rabbit aortic smooth muscle cells (RASMCs) cultured in serum-free medium for 24 hours. Induction of HO-1 mRNA was inhibited by actinomycin D and was associated with inhibition of RASMC proliferation. This probucol-induced increase in HO-1 mRNA and inhibition of RASMC proliferation was prevented by the HO inhibitor Sn(IV) protoporphyrin or transfection with small interference RNA (siRNA) to knockdown HO-1, but not by inactive Cu(II) protoporphyrin or scrambled siRNA. CONCLUSIONS: Probucol induces HO-1, and this contributes to the inhibition of vascular SMC proliferation. This novel finding may explain how probucol inhibits restenosis and highlights HO-1 as a target for therapeutic intervention against occlusive vascular disease. PMID- 15364807 TI - Circulating endothelial cell count as a diagnostic marker for non-ST-elevation acute coronary syndromes. AB - BACKGROUND: Shedding of endothelial cells from damaged endothelium into the blood occurs in a variety of vascular disorders. The purpose of this study was to evaluate the utility of circulating endothelial cell (CEC) count as a diagnostic marker of non-ST-elevation acute coronary syndromes (ACSs). METHODS AND RESULTS: CEC counts were determined immediately (H0), 4 hours (H4), and 8 hours (H8) after admission in 60 patients with documented non-ST-elevation ACS and 40 control patients with no evidence of coronary artery disease. A total of 32 patients in the ACS group had elevated CEC counts (>3 cells/mL) in relation to early admission and single-episode chest pain. Patients from the control group had normal CEC counts. The interval between the chest pain episode and elevation was significantly shorter for CEC than troponin I. No correlation was found between the 2 markers. Interestingly, a subgroup of ACS patients with initially normal troponin I levels had high CEC counts, thus allowing early diagnosis in 30% more cases. At H0, the mean area under the receiver operating characteristic curve was significantly higher with the CEC count than with the troponin I level. At H4 and H8, the combined use of CEC and troponin was significantly better as a marker of ACS than CEC alone or troponin I alone. CONCLUSIONS: This study demonstrates that CEC count can be used as an early, specific, independent diagnostic marker for non-ST-elevation ACS. A combined strategy using CEC count and troponin I level could provide an effective diagnostic tool. PMID- 15364808 TI - Pharmacological stabilization of mast cells abrogates late thrombotic events induced by diesel exhaust particles in hamsters. AB - BACKGROUND: Particulate air pollution is associated with cardiovascular diseases and myocardial infarction (MI). METHODS AND RESULTS: We investigated the relationship between airway inflammation and thrombosis 24 hours after intratracheal (IT) instillation of diesel exhaust particles (DEP; 50 microg/hamster). Mild thrombosis was induced in the femoral vein by endothelial injury, and the consequences of airway inflammation on thrombogenicity were studied via online video microscopy. Lung inflammation and histamine analysis in bronchoalveolar lavage (BAL) and plasma were performed after pretreatment with dexamethasone (DEX) or sodium cromoglycate (SC). DEP induced airway inflammation and histamine release in BAL and in plasma, and increased thrombosis, without elevating plasma von Willebrand factor (vWF) levels. The IT instillation of 400 nm positively charged polystyrene particles (500 microg/hamster), serving as particles that do not penetrate into the circulation, equally produced airway inflammation, histamine release, and enhanced thrombosis. Histamine in plasma resulted from basophil activation. Intraperitoneal (IP) pretreatment with DEX (5 mg/kg) abolished the DEP-induced histamine increase in BAL and plasma and abrogated airway inflammation and thrombogenicity. The IT pretreatment with DEX (0.5 mg/kg) showed a partial but parallel inhibition of all of these parameters. Pretreatment with SC (40 mg/kg, IP) strongly inhibited airway inflammation, thrombogenicity, and histamine release. CONCLUSIONS: Our results are compatible with the triggering of mast cell degranulation and histamine release by DEP. Histamine plays an initial central role in airway inflammation, further release of histamine by circulating basophils, and peripheral thrombotic events. Antiinflammatory pretreatment can abrogate the peripheral thrombogenicity by preventing histamine release from mast cells. PMID- 15364809 TI - Akt/protein kinase B and endothelial nitric oxide synthase mediate muscular neovascularization induced by tissue kallikrein gene transfer. AB - BACKGROUND: Angiogenesis gene therapy with human tissue kallikrein (hTK) has shown promise for ischemic disease. The present study was undertaken to (1) assess an optimal gene transfer modality, (2) clarify hTK angiogenic pathways, and (3) discount possible side effects. METHODS AND RESULTS: The hTK gene was transferred to murine adductors by increasing doses of an adenovirus (Ad.hTK). Heterologous protein production was evaluated by ELISA and immunohistochemistry. Structural and functional characteristics of hTK-induced neovascularization were assessed. Muscular endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF)-A mRNA and protein content were evaluated by real-time polymerase chain reaction and Western blotting. The ability of hTK to phosphorylate-activate Akt/protein kinase B (Akt-B) and VEGF receptor 2 (VEGF-R2) was also determined. Implication of the aforementioned mechanisms in Ad.hTK induced neovascularization was challenged by blocking Akt-B with a dominant negative Akt construct; NOS with N(G)-nitro-L-arginine methyl ester; and VEGF-A with neutralizing antibody, VEGF-R2 antagonist, or Ad carrying soluble VEGF-R1 gene. We found that 10(7) PFU Ad.hTK led to peak increases in capillary and arteriole density. Newly developed arterioles persisted for up to 8 weeks. Ad.hTK did not change microvascular permeability. Ad.hTK upregulated eNOS mRNA and protein and activated Akt-B through Ser-473 phosphorylation. Inhibitory studies documented that these biochemical events were instrumental to Ad.hTK-induced neovascularization. In contrast, Ad.hTK neither affected VEGF-A and VEGF-R2 levels nor increased VEGF-R2 phosphorylation. Consistently, Ad.hTK-induced neovascularization was not disturbed by any of the different approaches used to block VEGF-A. CONCLUSIONS: Our findings provide new information on the pathway involved in hTK-induced neoangiogenesis and represent an advancement toward clinical applications with Ad.hTK. PMID- 15364810 TI - Newly diagnosed and previously known diabetes mellitus and 1-year outcomes of acute myocardial infarction: the VALsartan In Acute myocardial iNfarcTion (VALIANT) trial. AB - BACKGROUND: A prior diagnosis of diabetes mellitus is associated with adverse outcomes after acute myocardial infarction (MI), but the risk associated with a new diagnosis of diabetes in this setting has not been well defined. METHODS AND RESULTS: We assessed the risk of death and major cardiovascular events associated with previously known and newly diagnosed diabetes by studying 14,703 patients with acute MI enrolled in the VALsartan In Acute myocardial iNfarcTion (VALIANT) trial. Patients were grouped by diabetic status: previously known diabetes (insulin use or diagnosis of diabetes before MI, n=3400, 23%); newly diagnosed diabetes (use of diabetic therapy or diabetes diagnosed at randomization [median 4.9 d after infarction], but no known diabetes at presentation, n=580, 4%); or no diabetes (n=10,719). Patients with newly diagnosed diabetes were younger and had fewer comorbid conditions than did patients with previously known diabetes. At 1 year after enrollment, patients with previously known and newly diagnosed diabetes had similarly increased adjusted risks of mortality (hazard ratio [HR] 1.43; 95% confidence interval [CI], 1.29 to 1.59 and HR, 1.50; 95% CI, 1.21 to 1.85, respectively) and cardiovascular events (HR, 1.37; 95% CI, 1.27 to 1.48 and HR, 1.34; 95% CI, 1.14 to 1.56). CONCLUSIONS: Diabetes mellitus, whether newly diagnosed or previously known, is associated with poorer long-term outcomes after MI in high-risk patients. The poor prognosis of patients with newly diagnosed diabetes, despite having baseline characteristics similar to those of patients without diabetes, supports the idea that metabolic abnormalities contribute to their adverse outcomes. PMID- 15364811 TI - Sensitivity and specificity of prenatal features of physiological shunts to predict neonatal clinical status in transposition of the great arteries. AB - BACKGROUND: Although prenatal diagnosis of transposition of the great arteries (TGA) reduces neonatal mortality, the preoperative course can be complicated in infants with a restrictive foramen ovale (FO) or a ductus arteriosus (DA) constriction. We sought to determine the specificity and sensitivity of prenatal features of physiological shunts in predicting postnatal clinical status in prenatally diagnosed TGA in babies delivered in a tertiary care center providing all facilities for neonatal urgent care. METHODS AND RESULTS: The outcomes of 130 fetuses with TGA were reviewed over a period of 5.5 years. Restriction of the FO and/or constriction of the DA could be analyzed in 119/130 fetuses at 36+/-2.7 weeks of gestation. Twenty-four out of 119 had at least 1 abnormal shunt (23 FO, 5 DA, and 4 both). Thirteen of 130 neonates had profound hypoxemia (PaO2<25 mm Hg) and metabolic acidosis (pH <7.15) in the first 30 minutes and required immediate balloon atrioseptostomy. Two who had abnormal FO and DA died despite aggressive resuscitation. The specificity and sensitivity of the fetal echo in predicting neonatal emergency were 84% and 54%, respectively. The specificity and sensitivity of a combination of restrictive FO and DA constriction were 100% and 31%, respectively. CONCLUSIONS: Restriction of the FO and/or of the DA has a high specificity to predict the need for emergency neonatal care in fetuses with TGA, but the sensitivity is too low to detect all high-risk fetuses. Exceptional procedures should be considered for fetuses that have a combination of restrictive FO and DA constriction. PMID- 15364812 TI - Circulating mononuclear cells in the obese are in a proinflammatory state. AB - BACKGROUND: In view of the increase in plasma concentrations of proinflammatory mediators tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and C reactive protein (CRP) in obesity, we investigated whether peripheral blood mononuclear cells (MNC) from obese subjects are in a proinflammatory state. METHODS AND RESULTS: MNC were prepared from fasting blood samples of obese (n=16; body mass index [BMI]=37.7+/-5.0 kg/m2) and normal-weight control (n=16; BMI=23.8+/-1.9 kg/m2) subjects. Nuclear factor kappaB (NF-kappaB) binding to DNA in nuclear extracts was elevated (P<0.05) and the inhibitor of NFkappaB-beta (IkappaB-beta) was significantly lower (P<0.001) in the obese group. Reverse transcription-polymerase chain reaction revealed elevated levels of migration inhibitor factor (MIF), IL-6, TNF-alpha, and matrix metalloproteinase-9 (MMP-9) mRNA expression in the obese subjects (P<0.05). Plasma concentrations of MIF, IL 6, TNF-alpha, MMP-9, and CRP were also significantly higher. Plasma glucose, insulin, and free fatty acids (FFAs) were measured, and homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Plasma FFA concentration related significantly to BMI, IL-6, and TNF-alpha mRNA expression and plasma CRP levels but not to HOMA-IR. On the other hand, the inflammatory mediators were significantly related to BMI and HOMA-IR. CONCLUSIONS: These data show (1) for the first time that MNC in obesity are in a proinflammatory state with an increase in intranuclear NF-kappaB binding, a decrease in IkappaB-beta, and an increase in the transcription of proinflammatory genes regulated by NF kappaB; (2) that plasma FFAs are a modulator of inflammation; and (3) that insulin resistance is a function of inflammatory mediators. PMID- 15364813 TI - Quantitative 3-dimensional echocardiography for accurate and rapid cardiac phenotype characterization in mice. AB - BACKGROUND: Insufficient techniques exist for rapid and reliable phenotype characterization of genetically manipulated mouse models of cardiac dysfunction. We developed a new, robust, 3-dimensional echocardiography (3D-echo) technique and hypothesized that this 3D-echo technique is as accurate as magnetic resonance imaging (MRI) and histology for assessment of left ventricular (LV) volume, ejection fraction, mass, and infarct size in normal and chronically infarcted mice. METHODS AND RESULTS: Using a high-frequency, 7/15-MHz, linear-array ultrasound transducer, we acquired ECG and respiratory-gated, 500-microm consecutive short-axis slices of the murine heart within 4 minutes. The short axis movies were reassembled off-line in a 3D matrix by using the measured platform locations to position each slice in 3D. Epicardial and endocardial heart contours were manually traced, and a B-spline surface was fitted to the delineated image curves to reconstruct the heart volumes. Excellent correlations were obtained between 3D-echo and MRI for LV end-systolic volumes (r=0.99, P<0.0001), LV end-diastolic volumes (r=0.99, P<0.0001), ejection fraction (r=0.99, P<0.0001), LV mass (r=0.94, P<0.0019), and infarct size (r=0.98, P<0.0001). Also, excellent correlations were found between the 3D-echo-derived LV mass and necropsy LV mass in normal mice (r=0.99, P<0.0001), as well as for 3D echo-derived infarct size and histologically determined infarct size (r=0.99, P<0.0001) in mice with chronic heart failure. Bland-Altman analysis showed excellent limits of agreement between techniques for all measured parameters. CONCLUSIONS: This new, fast, and highly reproducible 3D-echo technique should be of widespread applicability for high-throughput murine cardiac phenotyping studies. PMID- 15364814 TI - Relationship between cardiovascular risk as predicted by established risk scores versus plaque progression as measured by serial intravascular ultrasound in left main coronary arteries. AB - BACKGROUND: Intravascular ultrasound (IVUS) is increasingly used as an end point in studies aimed at reducing progression or inducing regression of coronary artery disease. However, data linking serial changes by IVUS with clinical outcomes are scarce. METHODS AND RESULTS: In the absence of a validated risk score for secondary prevention, we compared 3 established risk scores for primary prevention--PROCAM, SCORE, and Framingham--with plaque progression and lumen reduction as assessed with serial IVUS (follow-up, 18+/-9 months) in atherosclerotic left main coronary arteries of 56 patients with established atherosclerosis. For all 3 algorithms, patients at highest estimated risk of events showed greater plaque progression than patients at lowest risk (P<0.05 to <0.01). There were positive linear relationships between the risk of clinical events and plaque progression (r=0.41 to 0.60; P<0.002 to <0.0001). This translated into a greater decrease in lumen dimensions with increasing risk (P<0.05, PROCAM and SCORE). Risk prediction using the PROCAM algorithm showed the strongest relation with serial IVUS. During follow-up, 18 patients suffered from adverse cardiovascular events; these patients had an annual plaque progression that was significantly greater than other patients (25.2+/-19.4% versus 5.9+/ 15.6%, P<0.001). CONCLUSIONS: There was a positive linear relationship between the estimated risk of clinical events derived from all 3 established risk-score algorithms and the extent of plaque progression measured by serial IVUS. This translated into stenosis progression (reduction in lumen dimensions) with increasing clinical risk. PMID- 15364815 TI - "Early" class III drugs for the treatment of atrial fibrillation: efficacy and atrial selectivity of AVE0118 in remodeled atria of the goat. AB - BACKGROUND: Currently available antiarrhythmic drugs are only moderately effective against atrial fibrillation (AF) and may cause ventricular proarrhythmia. AVE0118 is a blocker of atrium-specific early K+ currents (I(Kur)/I(to)). METHODS AND RESULTS: Effects of intravenous AVE0118 and dofetilide on atrial effective refractory period (AERP) and inducibility of AF were measured before and after 48-hours of AF-induced electrical remodeling in the goat. During persistent AF (53+/-19 days), the cardioversion efficacy and effects on atrial wavelength of AVE0118, dofetilide, and ibutilide were evaluated. QT durations were measured during atrial pacing and persistent AF. After 48 hours of AF, the effect of dofetilide on AERP was reduced, and induction of AF was not prevented. In contrast, the class III action of AVE0118 was enhanced, and AF inducibility decreased from 100% to 32% (P<0.001). At 1, 3, and 10 mg x kg(-1) x h(-1), AVE0118 terminated persistent AF in 1 of 8, 3 of 8, and 5 of 8 goats, respectively. Dofetilide and ibutilide terminated AF in 1 of 5 and 2 of 7 goats. AVE0118 0.5, 1.5, and 5 mg/kg prolonged the AERP during AF and increased the fibrillation wavelength from 6.7+/-0.6 to 8.5+/-0.5, 9.7+/-0.5, and 11.2+/-0.9 cm (P<0.01). Whereas dofetilide and ibutilide prolonged QT duration, AVE0118 had no appreciable effect. CONCLUSIONS: AVE0118 markedly prolongs the AERP during AF without affecting QT duration. Cardioversion of AF was due to an approximately 2-fold increase in fibrillation wavelength. Atrium-selective class III drugs like AVE0118 may be a promising new option for safe and effective cardioversion of AF. PMID- 15364816 TI - Sensing tissue ischemia: another new function for capsaicin receptors? AB - BACKGROUND: Chest pain is a hallmark of myocardial ischemia, but its underlying signaling mechanisms remain poorly understood. The capsaicin receptor, vanilloid receptor-1 (VR1), is an important cation channel present on primary nociceptive neurons. We have shown that the VR1 is expressed on sensory nerve endings of the heart. In the present study, we determined the role of VR1s in activation of cardiac spinal afferent nerves caused by myocardial ischemia. METHODS AND RESULTS: Single-unit activity of cardiac afferents was recorded from the sympathetic chain of anesthetized ferrets. Cardiac afferents responded to 5 minutes of regional myocardial ischemia and topical application of 10 microg/mL bradykinin in a reproducible manner. Topical application of a specific VR1 antagonist, iodoresiniferatoxin (50 micromol/L), to the receptive field of afferents produced a large attenuation of the firing activity of cardiac afferents caused by myocardial ischemia. Iodoresiniferatoxin also significantly reduced the afferent response to bradykinin applied to the receptive field. Furthermore, treatment with a VR1 channel blocker, ruthenium red (200 micromol/L), had a similar inhibitory effect on the afferent responses to myocardial ischemia and bradykinin. CONCLUSIONS: This study provides the first functional evidence that ischemic stimulation of cardiac spinal afferent nerves is mediated through VR1s. The VR1 on the cardiac sensory nerve may function as a molecular sensor to detect tissue ischemia and activate cardiac nociceptors. PMID- 15364817 TI - Cholesteryl ester transfer protein and coronary artery disease: an observation with therapeutic implications. PMID- 15364818 TI - Targeting the chemokines in myocardial inflammation. PMID- 15364819 TI - Renin-angiotensin system and angiotensin receptor blockers in the metabolic syndrome. PMID- 15364820 TI - Cardiology patient page. Measurement of cholesterol: a patient perspective. PMID- 15364821 TI - Images in cardiovascular medicine. Ventricular tachycardia late after cardiac transplantation. PMID- 15364822 TI - Images in cardiovascular medicine. Multislice computed tomography in complex pulmonary atresia after stent implantation. PMID- 15364823 TI - Optimal implantation strategies using drug-eluting stents for in-stent restenosis: do we know the answer? PMID- 15364824 TI - Does the mitochondrial permeability transition have a role in preconditioning? PMID- 15364825 TI - Mortality risk and defibrillator benefit after myocardial infarction. PMID- 15364826 TI - Do meta-analyses of association studies of endothelial nitric oxide synthase variants and ischemic heart disease provide conclusive answers? PMID- 15364827 TI - Rhythm control and increased risk of noncardiovascular death in the Atrial Fibrillation Follow-up Investigation of Rhythm Management trial. PMID- 15364828 TI - Late coronary thrombosis secondary to a sirolimus-eluting stent. PMID- 15364829 TI - On-pump versus off-pump coronary artery bypass surgery in a matched sample of women: a comparison of outcomes. AB - BACKGROUND: Women have consistently higher mortality and morbidity than men after coronary artery bypass grafting (CABG). Whether elimination of cardiopulmonary bypass and performance of coronary artery bypass grafting off-pump (OPCAB) have a beneficial effect specifically in women has not been defined. METHODS AND RESULTS: From January 1998 through March 2002, 21 902 consecutive female patients at 82 hospitals underwent isolated CABG, as reported in an administrative database. Propensity score computer matching was performed based on 13 variables representing patient characteristics and preoperative risk factors to correct for and minimize selection bias. A total of 7376 (3688 pairs) women undergoing CABG surgery were able to be successfully matched. In a propensity score computer matched cohort, multivariate logistic regression (odds ratio) revealed that women undergoing on-pump surgery had a 73.3% higher mortality (P=0.002) and a 47.2% higher risk of bleeding complications (P=0.019). CONCLUSIONS: In a retrospective analysis of women undergoing CABG, computer-matched to minimize selection bias, off-pump surgery led to decreased mortality and morbidity including bleeding complications. PMID- 15364830 TI - Why is off-pump coronary surgery uncommon in Canada? Results of a population based survey of Canadian heart surgeons. AB - BACKGROUND: Off-pump coronary artery bypass (OPCAB) is proposed to improve clinical outcomes and decrease resource use. However, off-pump surgery is not widely used in Canada. The purpose of this study was to determine the current use of OPCAB in Canada and determine why surgeons have not adopted this technique. METHODS AND RESULTS: The study was a population-based survey of all adult Canadian cardiac surgeons in practice >1 year. Eligible division heads and surgeons were contacted by mail. Of 19 806 isolated coronary bypass surgeries performed by respondents in Canada last year, 3164 (16.0%) were performed off pump. More than 50% of Canadian surgeons performed OPCAB in <5% of coronary cases, and only 17% of surgeons performed OPCAB in >25% of coronary cases. Only 4 responding centers performed OPCAB in >25% of cases. Respondents were divided into those who performed <5% of cases off-pump (nonadopters), 5% to 25% off-pump (intermediate users), or >25% off-pump (enthusiasts). Mean number of distal anastomoses in off-pump cases were 1.7+/-0.6, 1.6+/-0.6, and 3.3+/-0.5 for nonadopters, intermediate users, and enthusiasts, respectively (P=0.001). Eleven percent of nonadopters, 55% of intermediate users, and 81% of enthusiasts believed OPCAB improved clinical outcomes (P<0.0001). Only 23% of all respondents felt OPCAB use would increase in the next 5 years. CONCLUSIONS: Concerns regarding incomplete revascularization and lack of proven clinical benefit have limited OPCAB to being performed routinely by only a small number of surgeons in Canada. PMID- 15364831 TI - Long-term outcome of isolated coronary artery bypass surgery in patients with severe left ventricular dysfunction. AB - BACKGROUND: Coronary artery bypass grafting (CABG) is indicated in patients with coronary artery disease and impaired ventricular function. However, earlier studies have suggested that prognosis of patients with severe left ventricular dysfunction is extremely poor. We used the APPROACH registry to derive contemporary estimates of prognosis associated with CABG for this high-risk patient population. METHODS AND RESULTS: The study group consisted of 7841 patients who had isolated CABG in the province of Alberta, Canada between 1996 and 2001. Patients with markedly reduced left ventricular function (ejection fraction [EF] <30%, Lo EF, n =430) were compared with those with moderate reduction in ventricular function (EF 30% to 50%, Med EF, n =2581) and those with normal left ventricular function (EF >50%, normal [Nl] EF, n=4830). The operative mortality was higher in the patient group with Lo EF (4.6%) compared with Med EF and Nl EF groups (3.4% and 1.9%, respectively, P<0.001). At 5 years, survival was 77.7% for Lo EF patients compared with 85.5% and 91.2% for Med EF and Nl EF patients, respectively (P<0.001). After controlling for other independent variables, the adjusted hazard ratio for death was 1.98 (95% CI, 1.49 to 2.62) for Lo EF relative to Nl EF. The mortality rate at 1 year was significantly lower for Lo EF patients who underwent CABG than it was for nonrevascularized Lo EF patients (risk-adjusted odds ratio, 0.36; 95% CI, 0.24 to 0.55). CONCLUSIONS: In the modern era of cardiac surgery, CABG can be performed in Lo EF cases with an acceptable perioperative mortality risk. Our estimate of 5-year survival in this high-risk group is better than previously reported in the literature from earlier periods. PMID- 15364832 TI - Extensive left ventricular remodeling does not allow viable myocardium to improve in left ventricular ejection fraction after revascularization and is associated with worse long-term prognosis. AB - BACKGROUND: Extensive left ventricular (LV) remodeling may not allow functional recovery after revascularization, despite the presence of viable myocardium. METHODS AND RESULTS: Seventy-nine consecutive patients with ischemic cardiomyopathy (left ventricle ejection fraction [LVEF] 29+/-7%) underwent surgical revascularization. Before revascularization, viability was assessed by metabolic imaging with F18-fluorodeoxyglucose and SPECT. LV volumes and LVEF were assessed by resting echocardiography. LVEF was re-assessed by echocardiography 8 to 12 months after revascularization. Three-year clinical follow-up (events: cardiac death, infarction, and hospitalization for heart failure) was also obtained. Forty-nine patients had substantial viability; 5 died before re assessment of LVEF. Of the remaining 44 patients, 24 improved > or =5% in LVEF after revascularization, whereas 20 did not improve in LVEF. LV end-systolic volume was the only parameter that was significantly different between the groups (109+/-46 mL for the improvers versus 141+/-31 mL for the nonimprovers; P<0.05). The change in LVEF after revascularization was linearly related to the baseline LV end-systolic volume, with a higher LV end-systolic volume associated with a low likelihood of improvement in LVEF after revascularization. During the 3-year follow-up, the highest event-rate (67%) was observed in patients without viable myocardium with a large LV size, whereas the lowest event rate (5%) was observed in patients with viable myocardium and a small LV size. Intermediate event rates were observed in patients with viable myocardium and a large LV size (38%), and in patients without viable myocardium and a small LV size (24%). CONCLUSIONS: Extensive LV remodeling prohibits improvement in LVEF after revascularization and affects long-term prognosis negatively, despite the presence of viability. PMID- 15364833 TI - Five-year angiographic patency of radial artery bypass grafts. AB - BACKGROUND: Little information exists regarding mid-term and long-term patency of radial artery grafts. METHODS AND RESULTS: We performed restudy coronary angiography at 5.2+/-0.4 years after surgery on 50 asymptomatic patients who had undergone coronary artery bypass graft surgery, using at least 1 radial artery graft, to determine both graft patency and presence of narrowing. We examined preoperative clinical or angiographic variables that might predict graft occlusion. Radial artery graft patency was 89%, with 91% of grafts free of narrowing. Preoperative New York Heart Association anginal class < or =2, target vessel proximal stenosis < or =70%, and small target vessel supply territory were predictive of graft occlusion. CONCLUSIONS: At 5 years after surgery, radial artery grafts have disease-free patency rates that are similar to other graft types. PMID- 15364834 TI - Single versus multiple internal mammary artery grafting for coronary artery bypass: 15-year follow-up of a clinical practice trial. AB - BACKGROUND: The long-term clinical advantages of using routine multiple internal mammary artery (IMA) grafts for coronary artery bypass (CAB) are not clear. This study was designed to test the hypothesis that multiple IMA grafts would provide better 15-year outcomes when compared with single IMA and vein grafts. METHODS AND RESULTS: Between 1984 and 1987, 1067 consecutive patients undergoing isolated CAB were referred to 1 surgeon practicing primarily single and another surgeon maximizing multiple IMA grafts (clinical practice trial). A 207-patient subset with multiple IMAs underwent postoperative graft angiography at 1 to 32 weeks to define initial IMA patency. Patients were followed-up yearly, and the groups were analyzed as (I) surgical strategy (surgeon operating) (single=413 versus multiple=654), (II) ultimate operation performed (single=418 versus multiple=449), or (III) single versus multiple coronary systems revascularized with IMAs (single=490 versus multiple=377). Advantages of this study design were that an entire referral population was examined, multiple IMAs were applied to the entire spectrum of baseline patient risk, 15-year follow-up provided a complete prognostic picture, and the subgroups were potentially comparable at baseline. In all 3 analyses, single and multiple groups were statistically similar with respect to baseline, operative, and immediate postoperative variables. Early IMA patency was 98.5% (333/338 grafts patent), validating the quality of IMA procedures. Unadjusted and adjusted 15-year outcome analyses for I, II, and III for death, myocardial infarction, percutaneous coronary intervention, redo coronary bypass, and the composite of all events identified multiple versus single as a significant predictor of outcome for the composite end point in adjusted analysis III (hazard ratio=0.808; 95% CI, 0.689 to 0.948; P=0.009), because of a 5% to 10% absolute reduction in each of the outcome variables at 15 years. Moreover, >50% reduction in reoperation rate was observed at 15 years in every analysis. CONCLUSIONS: At 15-year follow-up, multiple IMA grafting was associated with a 19.2% adjusted risk reduction in death and cardiac events, caused by decreases in all adverse end points and fewer reoperations. These data indicate that the clinical advantages of maximizing IMA conduits are significant. Based on this information, it is suggested that multiple IMA grafting to 2 coronary systems should be applied liberally to patients with noncardiac risk profiles predictive of long-term survival. PMID- 15364835 TI - Long-term patency of internal mammary artery bypass grafts: relationship with preoperative severity of the native coronary artery stenosis. AB - BACKGROUND: Internal mammary artery conduits (IMA) have an excellent long-term patency rate. Nevertheless, graft closure does occur and significantly limits future revascularization options. We sought to investigate the relationship between the long-term patency of IMA with clinical and angiographic parameters. Particularly, the preoperative degree of stenosis of the relevant bypassed coronary vessel was assessed to analyze the importance of chronic competitive flow on the arterial graft closure rate. METHODS AND RESULTS: Consecutive patients in whom occlusion of at least 1 IMA had been documented at angiography (OCC group) were compared with a group of patients with patent IMA grafts (PAT group). The degree of stenosis in the native coronary artery on which the IMA was placed was analyzed by off-line quantitative coronary angiography. Multivariate stepwise logistic regression was used to identify independent clinical and angiographic predictors of occlusion. The OCC group comprised 96 patients (67+/ 10 years) with 103 native bypassed arteries analyzed. The PAT group comprised 127 patients (69+/-8 years) with 170 native bypassed arteries analyzed. Both groups were similar except for gender (42% versus 32% female; P=0.04), height (166+/-8 versus 169+/-8 cm; P=0.006), minimum lumen diameter (0.76+/-0.7 versus 0.51+/ 0.5; P=0.001), and diameter stenosis of the native artery (73+/-25% versus 84+/ 16%; P<0.0001) in OCC versus PAT, respectively. In the multivariate analysis, only percent diameter stenosis was an independent and statistically significant predictor for graft patency. Among IMA placed on coronary arteries with a diameter of stenosis <50% (n=28), the occlusion rate was very high (79%). CONCLUSIONS: The degree of stenosis in the native vessel is a major predictor of internal mammary artery bypass graft patency. The association between nonsignificant stenosis of the native artery and high occlusion rate of the arterial bypass conduit raises concerns about the use of IMA in the treatment of native vessels with only mild or moderate stenosis. PMID- 15364836 TI - Effect of diabetes and associated conditions on long-term survival after coronary artery bypass graft surgery. AB - BACKGROUND: The effects of diabetes on short-term results of coronary artery bypass graft (CABG) surgery are known, but less is known about the long-term effects of diabetes and diabetic-related sequelae for patients undergoing this surgery. We studied the 10-year survival of nondiabetic and diabetic patients undergoing CABG surgery. METHODS AND RESULTS: A prospective regional cohort study was conducted of 36,641 consecutive isolated CABG patients in northern New England from 1992 through 2001. Patient records were linked to the National Death Index to assess mortality. There were 154,140 person-years of follow-up and 5779 deaths. Kaplan-Meier techniques were used. Survival was stratified into three categories: no diabetes, diabetes without peripheral vascular disease and renal failure, and diabetes with peripheral vascular disease and/or renal failure. The overall annual incidence rate of death was 3.7 deaths per 100 person-years. Annual incidence rates for nondiabetic subjects and diabetic subjects were similar: 3.1 deaths per 100 person-years and 4.4 deaths per 100 person-years, respectively. The annual incidence rate for diabetic subjects with renal failure, peripheral vascular disease, or both was 9.4 deaths per 100 person-years. The log rank test showed that the survival curves were significantly different (P<0.001). CONCLUSIONS: Patients that have diabetes without the sequelae of renal failure and/or peripheral vascular disease have long-term survival similar to but slightly less than patients without diabetes who undergo CABG surgery. Survival of CABG surgery patients with diabetes is greatly affected by associated comorbidities of peripheral vascular disease and renal failure. This knowledge may help guide the patient as well as the cardiologist and cardiac surgeon in making appropriate decisions in these critically ill patients. PMID- 15364837 TI - Statins are associated with a reduced incidence of perioperative mortality after coronary artery bypass graft surgery. AB - BACKGROUND: Statin therapy in nonsurgical patient populations is associated with a significant reduction in adverse cardiovascular events, including death, myocardial infarction (MI), and stroke. Recently, statin therapy was shown to be associated with a reduced incidence of postoperative mortality in patients undergoing major noncardiac vascular surgery. We investigated the influence of preoperative statin therapy on adverse outcomes after primary coronary artery bypass graft (CABG) surgery. METHODS AND RESULTS: A retrospective cohort study of patients undergoing primary CABG surgery with cardiopulmonary bypass (CPB) (n=1663) between January 1, 2000 and December 31, 2001 at the Texas Heart Institute was performed. Patients were classified into 2 groups: patients receiving preoperative statin therapy (n=943) and patients not receiving preoperative antihyperlipidemic therapy (n=720). To determine if preoperative statin therapy was independently associated with a reduction in the risk of adverse postoperative outcomes, multivariate stepwise logistic regression was performed controlling for patient demographics, medical history, and preoperative medications. Multivariate logistic regression analysis demonstrated that preoperative statin therapy was independently associated with a significant reduction ( approximately 50%) in the risk of 30-day all-cause mortality (3.75% versus 1.80%; P<0.05). The adjusted odds ratio for early mortality in patients receiving preoperative statin therapy compared with patients not receiving antihyperlipidemic agents was 0.53 (95% CI, 0.28 to 0.99). Statin therapy was not independently associated with a reduced risk of postoperative MI, cardiac arrhythmias, stroke, or renal dysfunction. In an attempt to further control for selection bias related to the choice of therapy, multivariate analysis of a propensity-matched cohort of 1362 patients revealed that preoperative statin therapy was independently associated with a significant reduction in the composite endpoint of 30-day all-cause mortality and stroke (7.1% versus 4.6%; P<0.05). CONCLUSIONS: Preoperative statin therapy may reduce the risk of early mortality after primary CABG surgery with CPB. PMID- 15364838 TI - Using the risk of restenosis as a guide to triaging patients between surgical and percutaneous coronary revascularization. AB - BACKGROUND: Coronary artery bypass graft (CABG) and percutaneous coronary revascularization (PCI) are strategies for treating coronary disease. Because the principal limitation of PCI is restenosis, CABG might be favored for those at high risk for restenosis. Using a clinical risk score for predicting restenosis, we examined whether patients with higher risks for restenosis were preferentially referred for CABG. METHODS AND RESULTS: A procedural registry of 2320 revascularization patients from whom data on procedure type, demographics, comorbid conditions, health status, vessel anatomy, and outcomes were taken was analyzed. Patients were classified and scored into 3 categories of restenosis risk ranging from 11% to 44%, as defined by 8 preprocedural characteristics. The objective of this study was to describe referral patterns between PCI and CABG in each category of risk. 2060 patients underwent nonemergent revascularization. 1404 of the patients underwent PCI and 656 were treated with CABG. Among the patients at low and intermediate risk for restenosis, twice as many were referred to PCI. Among those at the highest risk, 3-times as many were referred to PCI, resulting in a significant trend for those with the higher risks of restenosis to be preferentially referred to PCI (P=0.015). Similar results were seen when the analysis was restricted to only those with multivessel disease. CONCLUSIONS: Patients at higher risk for restenosis were being preferentially treated with PCI as opposed to CABG. These results may have implications for reevaluating current patterns of triaging patients between PCI and CABG, and for the use of drug eluting stents within PCI patients. PMID- 15364839 TI - Magnetic Vascular Port in minimally invasive direct coronary artery bypass grafting. AB - BACKGROUND: Minimally invasive direct coronary artery bypass grafting (MIDCAB) is a well-established operative procedure. However, it is technically demanding and is therefore somewhat underused. We evaluated the clinical and angiographic outcome of patients undergoing a MIDCAB procedure with the Ventrica Magnetic Vascular Port (MVP) system. METHODS AND RESULTS: A Ventrica MVP system was used in 10 of 11 selected MIDCAB patients. The system consists of 6 magnetic clips, with 3 clips forming a set. One magnetic clip set is positioned at the arteriotomy of the target artery and of the bypass graft using a preloaded delivery system. These ports then form an anastomosis by magnetic coupling. The mean age of the 10 patients (6 male) was 60.3+/-11.0 years. Three patients had an angiogram at the time of discharge and 8 returned for a 6-month angiogram. The total procedure time was 128.2+/-12.2 minutes. The mean anastomotic time was 199 seconds. The mean ischemic time during the anastomosis was 146+/-146 seconds. There were no in-hospital complications and no device-related adverse events. All 3 predischarge and all 8 6-month angiograms showed patent anastomoses. CONCLUSIONS: The magnetic vascular port facilitates the MIDCAB procedure significantly and reduces the ischemic time during the anastomosis. This minimally invasive procedure has the potential to be an alternative to percutaneous transluminal coronary angioplasty and stenting in proximal left anterior descending (LAD) stenosis. It may expand the acceptance of hybrid procedures in which a left internal mammary artery (LIMA)-to-LAD graft optimally supplies the anterior wall and the septum while the circumflex and right coronary artery may be treated interventionally. PMID- 15364840 TI - National survey of coronary artery bypass grafting for coronary stenosis caused by Kawasaki disease in Japan. AB - BACKGROUND: We surveyed the national experience of coronary artery bypass grafting (CABG) for coronary sequelae of Kawasaki disease (kDa) in 2002. METHODS AND RESULTS: A questionnaire was returned from 323/552 (59%) institutions. Two hundred forty-four patients (188 male 56 female) since 1975 were identified. The mean number of grafts was 1.8 and the age at operation ranged from 1 to 44 years (median 11 years). The interval from the onset of kDa to operation ranged from 1 month to 42 years (median 8 years), whereas the follow-up period was from 7 days to 25 years (median 5 years). Previous myocardial infarction was found in 70 patients (28%). When the age at operation was older than 12 years, patency rates for internal thoracic artery grafts (ITA) at 1, 5, and 15 years were 95%, 91%, and 91%, respectively (n=156). When the age at operation was younger than 12 years, the corresponding values were significantly lower, with 1, 5, and 15 years being 93%, 73%, and 65%, respectively (n=146) (P<0.05). Reoperation was performed in 14 patients (6%). Death occurred in 15 patients (6%). Nine of the 14 late deaths were sudden. Six of 8 patients with a left ventricular ejection fraction (LVEF) <40% died. CONCLUSIONS: The results of ITA in those aged 12 years or older were favorable. LVEF influenced prognosis. PMID- 15364841 TI - Aortic valve repair using a differentiated surgical strategy. AB - BACKGROUND: Reconstruction of the aortic valve for aortic regurgitation (AR) remains challenging, in part because of not only cusp or root pathology but also a combination of both can be responsible for this valve dysfunction. We have systematically tailored the repair to the individual pathology of cusps and root. METHODS: Between October 1995 and August 2003, aortic valve repair was performed in 282 of 493 patients undergoing surgery for AR and concomitant disease. Root dilatation was corrected by subcommissural plication (n=59), supracommissural aortic replacement (n=27), root remodeling (n=175), or valve reimplantation within a graft (n=24). Cusp prolapse was corrected by plication of the free margin (n=157) or triangular resection (n =36), cusp defects were closed with a pericardial patch (n=16). Additional procedures were arch replacement (n=114), coronary artery bypass graft (n=60) or mitral repair (n=24). All patients were followed-up (follow-up 99.6% complete), and cumulative follow-up was 8425 patient months (mean, 33+/-27 months).Results- Eleven patients died in hospital (3.9%). Nine patients underwent reoperation for recurrent AR (3.3%). Actuarial freedom from AR grade > or =II at 5 years was 81% for isolated valve repair, 84% for isolated root replacement, and 94% for combination of both; actuarial freedom from reoperation at 5 years was 93%, 95%, and 98%, respectively. No thromboembolic events occurred, and there was 1 episode of endocarditis 4.5 years postoperatively. CONCLUSIONS: Aortic valve repair is feasible even for complex mechanisms of AR with a systematic and individually tailored approach. Operative mortality is low and mid-term durability is encouraging. The incidence of valve related morbidity is low compared with valve replacement. PMID- 15364842 TI - Prospectively randomized evaluation of stented xenograft hemodynamic function in the aortic position. AB - BACKGROUND: Standard stented aortic xenograft valves have not yet been compared regarding their hemodynamic function using a stratified intraoperative randomization protocol. METHODS AND RESULTS: 100 patients were prospectively included after intraoperative metric sizing of the decalcified aortic annulus. They received Mosaic (M) or Perimount (P) aortic valve replacement. Patient age was 73+/-5 years, 51 were female, and New York Heart Association (NYHA) functional class was 2.8+/-0.5. The 21-mm annulus group consisted of 5 (M)/7 (P) patients, the 23-mm annulus group of 20 (M)/20 (P), the 25-mm annulus group of 18 (M)/19 (P), and the 27-mm annulus group of 4 (M)/7 (P) patients, respectively. Hemodynamic function was evaluated using transthoracic echocardiography before discharge and at follow-up (438+/-352 days). Surgery was uncomplicated in all patients. Labeled valve sizes were 0.93 (M) and 1.05 (P) mm smaller than the annulus diameters (p=NS). In-hospital mortality was 5%, all nonvalve-related. Transvalvular blood flow velocities and transvalvular pressure gradients were significantly lower in the 25 P versus the 25 M group at baseline and in the 23 P and 25 P groups at follow-up. There was a significant regression of left ventricular mass index in all patients at follow-up. However, left ventricular mass regression was more pronounced after P aortic valve replacement. CONCLUSIONS: Labeled sizes of prosthetic heart valves implanted are smaller than the true aortic annulus. Both standard aortic xenografts compared in this prospectively randomized trial provide a sufficient hemodynamic and functional outcome. PMID- 15364843 TI - Effects of paracommissural septal-lateral annular cinching on acute ischemic mitral regurgitation. AB - BACKGROUND: Previous experimental studies demonstrated that central septal lateral (SL) annular cinching (SLAC) abolishes acute ischemic mitral regurgitation (IMR), but whether localized cinching near the anterior (ACOM) or posterior (PCOM) commissure is equally effective is unknown. METHODS: Six adult sheep underwent implantation of 9 radiopaque markers on the left ventricle, 8 around the mitral annulus (MA) and 1 on each papillary muscle (PM) tip. Transannular SL sutures were placed at the valve center (CENT) and near ACOM and PCOM and externalized. Acute IMR was induced by proximal circumflex coronary snare occlusion. Biplane videofluoroscopy and transesophageal echocardiography were performed before and continuously during 3 episodes of myocardial ischemia including 20 seconds of SLAC at each different location. End-systolic MA SL dimension at each suture location and distances between the anterior and posterior PM tips and mid-septal annulus ("saddle horn") were calculated from the 3-dimensional (3D) marker coordinates. RESULTS: SLAC interventions in all 3 locations reduced the degree of IMR, but cinching at the center, SLAC(CENT), had a significantly greater effect on reducing the magnitude of IMR than SLAC(PCOM) or SLAC(ACOM) (mean grade of IMR reduction=1.0+/-0.5, 1.8+/-0.5, and 0.9+/-0.2 for SLAC(ACOM), SLAC(CENT), and SLAC(PCOM), respectively; P=0.044). Although ACOM and PCOM cinching reduced SL(CENT) somewhat, only SLAC(CENT) simultaneously reduced both SL(ACOM) and SL(PCOM) and also repositioned both PM tips closer to the annular saddle horn. CONCLUSIONS: SLAC in all 3 positions reduced acute IMR, but central SLAC cinching was most effective, reduced all mitral annular SL dimensions, and relocated both PM tips closer to the mid-septal annulus. Central SLAC is most capable of correcting the annular and subvalvular perturbations accompanying acute left ventricular ischemia that lead to IMR. PMID- 15364844 TI - Mechanism of recurrent ischemic mitral regurgitation after annuloplasty: continued LV remodeling as a moving target. AB - BACKGROUND: Patients who undergo ring annuloplasty for ischemic mitral regurgitation (MR) often have persistent or recurrent MR. This may relate to persistent leaflet tethering from left ventricle (LV) dilatation that is not relieved by ring annuloplasty. Therefore, the purpose of this study was to test the hypothesis that recurrent MR in patients after ring annuloplasty relates to continued LV remodeling. METHODS AND RESULTS: Serial echoes were reviewed in 30 patients (aged 72+/-11 years) who showed recurrent MR late (47+/-27 months) versus early (3.8+/-5.8 months) after ring annuloplasty for ischemic MR during coronary artery bypass grafting without interval infarction. Patients with intrinsic mitral valve disease were excluded. Echocardiographic measures of MR (vena contracta and jet area/left atrial area) and LV remodeling (LV dimensions, volumes, and sphericity) were assessed at each stage. The degree of MR increased from mild to moderate, on average, from early to late postoperative stages, without significant change in LV ejection fraction. Changes in MR paralleled increases in LV volumes and sphericity index at end-systole and end-diastole. The only independent predictor of late postoperative MR was LV sphericity index at end-systole. CONCLUSIONS: Recurrent MR late after ring annuloplasty is associated with continued LV remodeling, emphasizing its dynamic relation to the LV. PMID- 15364845 TI - Cutting second-order chords does not prevent acute ischemic mitral regurgitation. AB - BACKGROUND: Cutting anterior mitral leaflet second-order chordae has been proposed for repair in ischemic mitral regurgitation (IMR). We examined the efficacy of such chordal cutting in preventing acute IMR. METHODS AND RESULTS: Six sheep underwent radiopaque marker placement (left ventricle, mitral annulus, papillary muscles [PMs], and leaflets). The largest second-order chord from each PM was encircled with exteriorized wire snares. Three-dimensional marker coordinates were obtained with biplane videofluoroscopy before and during acute ischemia (80 seconds of mid-circumflex occlusion). Color Doppler transesophageal echocardiography was used to grade MR on a 0 to 4+ scale. Data were acquired immediately before and after dividing second-order chordae. Slope of the end diastolic volume-stroke work relationship (PRSW) was calculated to assess systolic function. Chordal cutting increased anterior leaflet inflection angle (155+/-12 versus 162+/-9 degrees; P=0.03), resulting in a flatter leaflet, but did not increase effective leaflet length (1.97+/-0.24 versus 2.08+/-0.23 cm; P=0.15); PRSW decreased (63+/-15 versus 56+/-12 mm Hg; P=0.008). Both before and after chordal cutting, ischemia caused: Septal-lateral annular dilation (P=0.005), posterior PM displacement away from the mid-septal annulus (P=0.06), increased leaflet tenting area (P=0.001), and increased leaflet tenting volume (P=0.002). Before chordal cutting, MR increased significantly during ischemia (0.5+/-0.3 versus 1.7+/-0.4; P<0.001), and IMR increased similarly even after the second-order chords were cut (0.7+/-0.4 versus 1.9+/-0.9; P<0.001). CONCLUSIONS: Cutting second-order chordae resulted in LV systolic dysfunction and neither prevented nor decreased the severity of acute IMR, septal-lateral annular dilation, leaflet tenting area, or leaflet tenting volume. PMID- 15364846 TI - Undersized mitral annuloplasty alters left ventricular shape during acute ischemic mitral regurgitation. AB - BACKGROUND: Underlying left ventricular (LV) dysfunction contributes to poor survival after operation to correct ischemic mitral regurgitation (IMR). Many surgeons do not appreciate that a key component of the Bolling undersized mitral ring annuloplasty concept is to decrease LV wall stress by altering LV shape, but precise 3-dimensional (3-D) geometric data do not exist substantiating this effect. We tested the hypothesis that annular reduction decreases regional circumferential LV radius of curvature (ROC) in a model of acute IMR. METHODS: Eight adult sheep underwent insertion of an adjustable Paneth-type annuloplasty suture and radiopaque markers on the LV and mitral annulus. The animals were studied with biplane videofluoroscopy during baseline conditions, then before and after tightening the annuloplasty suture during proximal left circumflex occlusion. End-systolic circumferential regional LV ROC and mitral annular area were computed. RESULTS: Acute IMR was eliminated (MR grade 2.1+/-0.4 to 0.4+/ 0.4, mean+/-SD, P<0.05) by tightening the Paneth annuloplasty suture. Paneth suture tightening during circumflex occlusion also decreased end-systolic regional circumferential radii of curvature at the basal (anterior, 3.40+/-0.16 to 3.34+/-0.14 cm; posterior, 3.31+/-0.23 to 3.24+/-0.26 cm; P<0.05) and equatorial levels (anterior, 2.99+/-0.21 to 2.89+/-0.29 cm; posterior, 2.86+/ 0.38 to 2.81+/-0.41 cm; P<0.05). CONCLUSIONS: Acute proximal circumflex occlusion caused IMR and increased end-systolic LV radii of curvature in this experimental preparation. Annular reduction sufficient to abolish IMR also decreased end systolic anterior and posterior LV ROC, which would be expected to reduce LV wall stress and oxygen consumption in these regions, both potentially beneficial effects. The long-term effects of undersized annuloplasty on LV remodeling and function, however, will require further study in chronic animal preparations or patients with chronic IMR. PMID- 15364847 TI - Restrictive annuloplasty and coronary revascularization in ischemic mitral regurgitation results in reverse left ventricular remodeling. AB - BACKGROUND: Data on combined coronary artery bypass grafting (CABG) and restrictive annuloplasty in patients with ischemic cardiomyopathy are scarce, and the effect on reverse left ventricular (LV) remodeling is unknown. METHODS AND RESULTS: 51 patients with ischemic LV dysfunction (LV ejection fraction 31+/-8%) and severe mitral regurgitation (grade 3 to 4+) underwent CABG and restrictive annuloplasty with stringent downsizing of the mitral annulus (by 2 sizes, Physio ring, mean size 28+/-2). Serial transthoracic echocardiographic studies were performed (before surgery and within 3 months and 1.5 years after surgery) to assess mitral regurgitation, transmitral gradient, leaflet coaptation, and left atrial and LV reverse remodeling. Clinical follow-up (New York Heart Association [NYHA] class, survival, events) was assessed at 2-year follow-up. Early operative mortality was 5.6%; at 2-year follow-up, all patients were free of endocarditis and thromboembolism, and 1 needed re-operation for recurrent mitral regurgitation; 2-year survival was 84%. NYHA class improved from 3.4+/-0.8 to 1.3+/-0.4 (P<0.01), with all patients in class I/II. Intraoperative transesophageal echo showed minimal (grade 1+) mitral regurgitation in 8 patients and none in 43, without stenosis. Leaflet coaptation was 0.8+/-0.2 cm. These values remained unchanged; all patients had no or minimal (grade 1+) mitral regurgitation at 2-year follow-up. LV end-systolic and end-diastolic dimensions decreased from 51+/-10 to 43+/-12 mm (P<0.001) and from 64+/-8 to 58+/-11 mm (P<0.001). Left atrial dimension decreased from 53+/-8 to 47+/-7 mm (P<0.001). CONCLUSIONS: Excellent results of combined restrictive annuloplasty and CABG were obtained. Residual mitral regurgitation was absent/minimal at 2-year follow-up, associated with a significant reduction in left atrial dimension and LV reverse remodeling. PMID- 15364848 TI - Alterations in left ventricular torsion and diastolic recoil after myocardial infarction with and without chronic ischemic mitral regurgitation. AB - BACKGROUND: Chronic ischemic mitral regurgitation (CIMR) is associated with heart failure that continues unabated whether the valve is repaired, replaced, or ignored. Altered left ventricular (LV) torsion dynamics, with deleterious effects on transmural gradients of oxygen consumption and diastolic filling, may play a role in the cycle of the failing myocardium. We hypothesized that LV dilatation and perturbations in torsion would be greater in animals in which CIMR developed after inferior myocardial infarction (MI) than in those that it did not. METHODS: 8+/-2 days after marker placement in sheep, 3-dimensional fluoroscopic marker data (baseline) were obtained before creating inferior MI by snare occlusion. After 7+/-1 weeks, the animals were restudied (chronic). Inferior MI resulted in CIMR in 11 animals but not in 9 (non-CIMR). End-diastolic septal-lateral and anterior-posterior LV diameters, maximal torsional deformation (phi(max), rotation of the LV apex with respect to the base), and torsional recoil in early diastole (phi(5%), first 5% of filling) for each LV free wall region (anterior, lateral, posterior) were measured. RESULTS: Both CIMR and non-CIMR animals demonstrated derangement of LV torsion after inferior MI. In contrast to non CIMR, CIMR animals exhibited greater LV dilation and significant reductions in posterior maximal torsion (6.1+/-4.3 degrees to 3.9+/-1.9 degrees * versus 4.4+/ 2.5 degrees to 2.8+/-2.0 degrees; mean+/-SD, baseline to chronic, *P<0.05) and anterior torsional recoil (-1.4+/-1.1 degrees to -0.2+/-1.0 degrees versus -1.2+/ 1.0 degrees to -1.3+/-1.6 degrees ). CONCLUSIONS: MI associated with CIMR resulted in greater perturbations in torsion and recoil than inferior MI without CIMR. These perturbations may be linked to more LV dilation in CIMR, which possibly reduced the effectiveness of fiber shortening on torsion generation. Altered torsion and recoil may contribute to the "ventricular disease" component of CIMR, with increased gradients of myocardial oxygen consumption and impaired diastolic filling. These abnormalities in regional torsion and recoil may, in part, underlie the "ventricular disease" of CIMR, which may persist despite restoration of mitral competence. PMID- 15364849 TI - Importance of mitral valve second-order chordae for left ventricular geometry, wall thickening mechanics, and global systolic function. AB - BACKGROUND: Mitral valvular-ventricular continuity is important for left ventricular (LV) systolic function, but the specific contributions of the anterior leaflet second-order "strut" chordae are unknown. METHODS AND RESULTS: Eight sheep had radiopaque markers implanted to silhouette the LV, annulus, and papillary muscles (PMs); 3 transmural bead columns were inserted into the mid lateral wall between the PMs. The strut chordae were encircled with exteriorized wire snares. Three-dimensional marker images and hemodynamic data were acquired before and after chordal cutting. Preload recruitable stroke work (PRSW) and end systolic elastance (E(es)) were calculated to assess global LV systolic function (n=7). Transmural strains were measured from bead displacements (n=4). Chordal cutting caused global LV dysfunction: E(es) (1.48+/-1.12 versus 0.98+/-1.30 mm Hg/mL, P=0.04) and PRSW (69+/-16 versus 60+/-15 mm Hg, P=0.03) decreased. Although heart rate and time from ED to ES were unchanged, time of mid-ejection was delayed (125+/-18 versus 136+/-19 ms, P=0.01). Globally, the LV apex and posterior PM tip were displaced away from the fibrous annulus and LV base-apex length increased at end-diastole and end-systole (all +1 mm, P<0.05). Locally, subendocardial end-diastolic strains occurred: Longitudinal strain (E22) 0.030+/ 0.013 and radial thickening (E33) 0.081+/-0.041 (both P<0.05 versus zero). Subendocardial systolic shear strains were also perturbed: Circumferential longitudinal "micro-torsion" (E12) (0.099+/-0.035 versus 0.075+/-0.025) and circumferential radial shear (E13) (0.084+/-0.023 versus 0.039+/-0.008, both P<0.05). CONCLUSIONS: Cutting second-order chords altered LV geometry, remodeled the myocardium between the PMs, perturbed local systolic strain patterns affecting micro-torsion and wall-thickening, and caused global systolic dysfunction, demonstrating the importance of these chordae for LV structure and function. PMID- 15364850 TI - Extending the boundaries of the primary arterial switch operation in patients with transposition of the great arteries and intact ventricular septum. AB - BACKGROUND: We have previously suggested that the primary arterial switch operation is a feasible strategy for patients with transposition of the great arteries and intact ventricular septum (TGA-IVS) up to age 2 months. This study reports our current results with this approach and examines whether this policy could be extended beyond age 2 months. METHODS AND RESULTS: 380 patients who underwent arterial switch for TGA-IVS were reviewed. 275 patients were younger than 3 weeks at the time of surgery (early switch group); 105 patients were 3 weeks or older (range, 21 to 185 days) (late switch group). There was no difference in outcome in terms of in-hospital mortality (5.5% versus 3.8%) or need for mechanical circulatory support (3.6% versus 5.7%) between early and late switch groups. However, duration of postoperative ventilation (4.9 versus 7.1 days, P=0.012) and length of postoperative stay (12.5 versus 18.9 days, P<0.001) were significantly prolonged in the late switch group. Primary left ventricular failure resulting in death occurred in 2 patients in the late switch group, with no deaths in 9 patients aged 2 to 6 months. CONCLUSIONS: This experience confirms that in TGA-IVS, the left ventricle maintains the potential for systemic work well beyond the first month of life. Consequently, neonates at high risk or late referrals can benefit from delayed arterial switch, even beyond age 2 months. However, the need for mechanical support in some of the older patients may limit the widespread adoption of such a strategy. PMID- 15364851 TI - Long-term predictors of aortic root dilation and aortic regurgitation after arterial switch operation. AB - BACKGROUND: Neo-aortic root dilation (ARD) and neo-aortic regurgitation (AR) may be progressive after arterial switch operation (ASO) for d-loop transposition of the great arteries (dTGA). We sought to identify predictors of ARD and AR after ASO. METHODS AND RESULTS: 335 patients were identified who underwent ASO for dTGA with intact ventricular septum or ventricular septal defect (VSD), including double-outlet right ventricle (DORV), before 2001 with at least 1 postoperative echocardiogram at our institution, at least 1 year after ASO, and no previous atrial switch procedure (median follow-up of 5.0 years). Probability of freedom from ARD was 97%, 92%, 82%, and 51%, from at least moderate AR was 98%, 97%, 96%, and 93%, and from neo-aortic valve or root surgery was 100%, 100%, 99%, and 95%, at 1, 2, 5, and 10 years, respectively. For patients in whom ARD developed, progressive dilation was not observed during late follow-up. By Kaplan-Meier method, independent predictors of ARD, with neo-aortic root z-score of > or =3.0, were previous pulmonary artery band (PAB) (P=0.002, hazard ratio [HR]=2.4) and later time period when ASO was performed (P<0.002, HR=19.0). Risk factor for at least moderate AR was age > or =1 year at ASO (P=0.002, HR=5.8), which was closely related to VSD repair at ASO (P<0.001) and previous PAB. CONCLUSIONS: Significant ARD and AR continue to develop over time after ASO, but ARD does not tend to be progressive during late follow-up. Previous PAB was a significant risk factor for ARD. Older age at time of ASO, presence of VSD, and previous PAB were risk factors for AR. PMID- 15364852 TI - Prevention of early sudden circulatory collapse after the Norwood operation. AB - BACKGROUND: After modifications in our perioperative management protocol, we have observed a decrease in sudden circulatory collapse after the Norwood operation. The current study examines early outcomes after the Norwood operation in our unit in an attempt to identify variables that may have altered the risk of unexpected circulatory collapse. METHODS AND RESULTS: We studied 105 consecutive neonates who underwent a Norwood operation in our institution. Our treatment protocol has changed in the past 3 years to include the use of alpha-blockade with phenoxybenzamine (POB) for systemic afterload reduction and selective cerebral perfusion. Forty-eight infants had selective cerebral perfusion. Forty-two infants received POB. Sixty patients had hypoplastic left heart syndrome. There was no difference in age, diagnosis, number of neonates with weight <2.5 kg, aortic size diameter <2 mm, highest preoperative lactate level, and shunt size indexed to body weight among patients with or without use of POB. Twenty-five infants had circulatory collapse during the first 72 hours. Twelve of them could be explained by technical issues. Thirteen others who appeared clinically stable had early sudden circulatory collapse without an apparent cause. Sixteen out of 25 neonates died. Of those with technical problems, 8 out of 12 died. Based on the hazard function, 3 incremental risk factors for early circulatory collapse were technical issue at operation (P<0.001), longer cross-clamp time (P<0.007), and no use of POB (P<0.002). For a technically successful operation, freedom from circulatory collapse at 72 hours is 95% with the use of POB versus 69% without (P<0.002). Diagnosis, aortic size, atrioventricular valve function, birth weight, age at operation, and total circulatory arrest time and were not predictive of early sudden circulatory collapse. CONCLUSIONS: Recent changes in our treatment protocol have resulted in a decrease incidence of sudden circulatory collapse after the Norwood operation. Optimal surgical technique is the most important predictor of early survival. The use of aggressive afterload reduction with POB reduced the risk of early sudden arrest. PMID- 15364853 TI - Hypoplasia of the small pulmonary arteries in hypoplastic left heart syndrome with restrictive atrial septal defect. AB - BACKGROUND: Restrictive atrial septal defect (ASD) (including intact atrial septum [IAS]) has been reported to be a risk factor that negatively impacts survival in hypoplastic left heart syndrome (HLHS). Although lymphangiectasia and "arterialization" of the veins of the lung in HLHS with restrictive ASD have been reported, they cannot fully explain the high mortality. We have introduced a new method of evaluating the development of the pulmonary vasculature in histological sections and used it to assess patients' lungs. We tested the hypothesis that the small pulmonary arteries (SPA), which are pulmonary arteries in a histological section whose radii are approximately 25 microm to 250 microm, in HLHS with restrictive ASD are hypoplastic, but that the alveoli are not, to elucidate the mechanism underlying the poor outcome of these patients. METHODS AND RESULTS: Fourteen HLHS patients between 1 day and 40 days of age were studied. In 8 cases, the ASD was restrictive [R(+) group], and in the other 6 cases it was not [R(-) group]. Specimens from 12 autopsies of cases with no congenital heart or pulmonary disease were examined as a control group (C group). As a novel histological parameter, we assessed the size of SPA in relation to the size of accompanying bronchioles to identify SPA underdevelopment. To evaluate the development of alveoli and interstitial tissue, radial alveolar counts (RAC), which reflect alveolar maturity and complexity, were also performed. Statistical comparisons between groups were made by analysis of covariance with age as a covariant factor. When the radius of the accompanying bronchiole was 100 microm, the radius of the SPA was 34.0+/-10.8 microm in the R(+) group, and significantly lower than the 46.6+/-8.5 microm in R(-) group (P=0.0022) and 70.5+/-8.4 microm in the C group (P<0.0001). The RAC was in 3.5+/-0.9 in the R(+) group, 3.4+/-0.6 in the R(-) group, and 3.7+/-0.9 in the C group (no significant differences between groups). CONCLUSIONS: The SPA in HLHS with restrictive ASD were underdeveloped compared with the SPA in HLHS with nonrestrictive ASD and the controls, but their alveoli were not hypoplastic. Based on these results, it is speculated that SPA hypoplasia may be responsible for the poor outcome of HLHS with restrictive ASD. PMID- 15364854 TI - Brain versus lung: hierarchy of feedback loops in single-ventricle patients with superior cavopulmonary connection. AB - BACKGROUND: CO2 vasodilates and O2 vasoconstricts the cerebral vascular bed; the opposite is true in the lungs. When the brain and lungs are connected exclusively in series, which feedback loop predominates is unknown. The circulation of the superior cavopulmonary connection (SCPC) provides a unique physiology to answer this question. METHODS AND RESULTS: To determine cerebral and pulmonary blood flow and to establish the hierarchy of cerebral and pulmonary feedback mechanisms, 12 intubated, ventilated, single-ventricle patients in SCPC physiology (age 2.2+/-0.5 years) underwent magnetic resonance imaging velocity mapping of their jugular veins and aorta in room air, hypercarbia, and 100% O2. Flows in these vessels and arterial blood gases were measured. With 22+/-6 torr CO2 (Pco2) increased from 40 to 63 mm Hg, P<0.01), flow to the brain and lungs increased (1.5 to 2.7 L/min per m2, P=0.0003), Po2 improved (48 to 60 mm Hg, P=0.0004), and cardiac index increased (4.3 to 5.4 L/min per m2, P=0.0003). The increased cardiac index accounted for the increased cerebral and pulmonary blood flow (R=0.73, P=0.02) and cerebral O2 transport increased by 80% (P=0.0005) while preserving body O2 delivery. Hyperoxia did not change cerebral and pulmonary blood flow; Po2 increased 94% (P=0.01). CONCLUSIONS: The cerebral CO2 feedback loop predominates over the pulmonary one when they directly compete with each other. CO2 has a major impact on flow distribution whereas O2 has little impact. Increased CO2 improves cerebral oxygenation in SCPC patients. This may provide a clue in determining neurological sequelae in SC physiology and may influence timing of Fontan completion. PMID- 15364855 TI - Pulmonary regurgitation is an important determinant of right ventricular contractile dysfunction in patients with surgically repaired tetralogy of Fallot. AB - BACKGROUND: Evaluation of right ventricular (RV) function in patients with pulmonary regurgitation (PR) after tetralogy repair remains challenging because of abnormal RV loading conditions. METHODS AND RESULTS: We examined 124 patients, aged 21+/-11.4 years, who had tetralogy repair at 3.7+/-3.5 years. By Doppler echocardiography, 33 patients had mild, 22 moderate, and 69 severe PR; 55 had significant tricuspid regurgitation (TR). Myocardial velocities, myocardial acceleration during isovolumic contraction (IVA), strain, and strain rate were measured at RV and LV base. Tricuspid valve annulus was measured in a 4-chamber view. QRS, QT, and JT intervals and their dispersions were measured from 12-lead electrocardiogram. IVA in the RV was lower in all patients compared with controls (0.8+/-0.4 versus 1.8+/-0.5, P<0.0001) and correlated with the severity of PR (r= 0.43, P<0.0001), whereas myocardial velocities, and strain/strain rate did not. LV IVA correlated with PR (r=-0.32, P<0.001) and with RV IVA (r=0.28, P<0.003). Patients with severe PR had a higher incidence of TR (r=0.69, P<0.0001) and lower RV IVA (1.0+/-0.4 versus 0.6+/-0.3, P<0.0001), a larger tricuspid valve ring diameter (P<0.0001), and prolonged electrical depolarization (P<0.001). Age at surgery or examination did not correlate with PR and with RV function assessed by IVA. In the RV, IVA correlated inversely with QRS duration (P<0.01). CONCLUSIONS: Although load-dependent myocardial velocities and strain are not influenced by the severity of PR and presence of significant TR, IVA demonstrates reduced contractile function in relation to the degree of PR and may be an early, sensitive index for selecting patients for valve replacement. PMID- 15364856 TI - Early clinical results of the telemetric adjustable pulmonary artery banding FloWatch-PAB. AB - BACKGROUND: Adjustment of pulmonary artery banding (PAB) may be a challenging procedure in complex congenital heart defects. Whatever the technique used, subsequent re-operations are frequently needed to control the pulmonary blood flow or pressures. OBJECTIVE: To report the efficacy of a new telemetric adjustable PAB (FloWatch-PAB) operated with the help of an external control unit that transmits to the implant energy and commands to further narrow or release the pulmonary artery using radiofrequency waves. METHODS AND RESULTS: In a multicenter, prospective, nonrandomized, single-arm clinical investigation, 13 children (median age, 4.5 months; range, 6 days to 11 years; median weight, 4.2 kg; range, 3.1 to 27 kg) underwent implantation of the FloWatch-PAB through median sternotomy in 8 and left thoracotomy in 5. The diagnosis was multiple ventricular septal (VSD) defects with complex anatomy in 3, single ventricle without pulmonary stenosis in 2, VSD with elevated pulmonary vascular resistance (PVR) in 2, atrio-ventricular canal (AVC) with elevated pulmonary vascular resistance in 1, AVC with diminutive right ventricle in 1, complex transposition of the great arteries in 3, and pulmonary atresia with complex pulmonary arteries anatomy in 1. All patients had normosystemic systolic pulmonary artery pressure. Additional procedures were performed in 7: atrial septectomy in 2, double aortic arch division in 1, patent ductus arteriosus ligation in 2, and coarctation repair in 2. There were no early or late deaths or device-related complications in a mean follow-up of 24 weeks (range, 18 to 42 weeks). A mean of 5.8 telemetric regulations per patient using the FloWatch-PAB were required to adjust the tightening of the PAB to the clinical needs (narrowing 74%, releasing 26%). At last follow-up, systolic pulmonary artery pressure was within normal range in all patients but 1. Systemic oxygen saturation demonstrated optimal regulation of the pulmonary blood flow in all according to each specific defect. Four patients were successfully corrected (VSD closure, AVSD repair, and 2 arterial switches with VSD closure). The device was easily removed and the pulmonary artery re-expanded spontaneously. CONCLUSIONS: This new device is safe and allows optimal adjustment of PAB in complex heart defects. In children requiring PAB, the use of this technology can obviate the need for early re-operations and appears to be a valuable option in the panel of surgical alternatives for selected infants. PMID- 15364857 TI - Atrial tachyarrhythmias after the maze procedure: incidence and prognosis. AB - BACKGROUND: The Maze procedure restores normal sinus rhythm in the majority of patients. However, atrial tachyarrhythmias (ATA) are a common early complication after the operation. The purpose of this study was to define the incidence and natural history of ATA after the Maze procedure. METHODS: Complete medical records from 200 patients who underwent the Maze procedures (I, II, and III) from 1987 to 2002 were examined for all episodes of early postoperative ATA that occurred during the first 30 days after the procedure. Two electrophysiologists independently reviewed all postoperative 12-lead electrocardiograms. RESULTS: ATA occurred in 86 patients (43%) after the Maze procedure. Of the patients with ATA, 59% had atrial fibrillation (AF), 14% had atrial flutter (AFL), and 27% had both AF and AFL. Of the patients with AF or AFL, 20% and 5%, respectively, also had episodes of atrial tachycardia and supraventricular tachyarrhythmia. The peak incidence of early postoperative ATA was on postoperative day 8. The average duration of ATA was 5.7+/-5.0 days. Late recurrence of AF (>1 year postoperatively) occurred in 7.0% of patients who had early postoperative ATA and 8.8% of patients without early postoperative ATA (P=0.8). CONCLUSIONS: ATA occurred in 43% of patients after the Maze procedure. The tachyarrhythmias occurred primarily within 8 days after surgery and resolved within 3 weeks in almost all patients. There was no relationship between the incidence of early postoperative ATA and the late recurrence of AF. PMID- 15364858 TI - Safety and efficacy of surgical ventricular restoration in unstable patients with recent anterior myocardial infarction. AB - BACKGROUND: The effects and efficacy of surgical ventricular restoration (SVR) in ischemic cardiomiopathy caused by chronic anterior myocardial infarction (MI) are well established. Normally, SVR is delayed at least 3 months after MI to allow the healing of infarcted tissue. Some patients have instability <30 days after anterior MI, with increased risk for morbidity and mortality.Objectives- This study tests the safety and efficacy of SVR in the setting of subacute complicated anterior MI, in terms of early and late outcome. METHODS AND RESULTS: 74 patients (62+/-10 years) were submitted to SVR at < or =30 days after anterior MI for clinical instability and were retrospectively selected from a series of 430 patients undergoing SVR at our center, between 1998 and 2001. The surgical indications included: angina (60%); New York Heart Association class 4 (62%); clinical signs of heart failure (18%); life-threatening arrhythmias (12%); and cardiogenic shock in 4% (or 3) patients. Follow-up is available for 93% of patients. All patients had coronary artery bypass grafting (CABG) (3.1+/-1.2) with internal mammary artery (IMA) utilization. An endoventricular patch was used in 17 patients (23%); direct ventriculotomy closure was used in the remaining patients. Operative mortality was 5.4% (4/74). Hemodynamic parameters improved significantly in patients with dilated hearts and reduced ejection fraction. Mitral regurgitation that resulted was significantly reduced. Survival at 3 years was 87% in the overall population and 85% in patients 70 years or older. CONCLUSIONS: This study reports the largest series of patients with complicated, recent anterior MI treated with SVR. The results show that SVR is feasible, has acceptable in-hospital mortality, and has good early and late outcome. Further experience is needed to establish whether SVR, which excludes the infarcted region, can prevent the long-term adverse remodeling of LV dilated hearts after anterior infarction. PMID- 15364859 TI - Atrial natriuretic peptide helps prevent late remodeling after left ventricular aneurysm repair. AB - BACKGROUND: Left ventricular aneurysm repair (LVR) reduces LV wall stress and improves LV function. However, as we reported previously, the initial improvement of LVR was short-term because of LV remodeling but could be maintained longer with postoperative use of an angiotensin-converting enzyme (ACE) inhibitor. Atrial natriuretic peptide (ANP) has been used to treat patients with heart failure by natriuretic and vasodilatory actions. Recent reports have suggested that ANP inhibits the rennin-angiotensin system. In this study, the effects of ANP after LVR were evaluated. METHODS AND RESULTS: Rats that had an LV aneurysm 4 weeks after left anterior descending artery ligation underwent LVR by plicating the LV aneurysm and were randomized into 2 groups: LVR+A group was intravenously administrated with 10 microg/h of carperitide, recombinant alpha-hANP, by osmotic pump for 4 weeks, and the LVR group was given normal saline. Echocardiography revealed better LV remodeling and function in LVR+A group than in LVR group. Four weeks after LVR, left ventricular end diastolic pressure (LVEDP) and Tau were significantly lower in LVR+A group (LVEDP: 10+/-4 in LVR+A group versus 18+/-6 mm Hg in LVR group, Tau: 13+/-2 versus 17+/-2ms). End-systolic elastance (Ees) was higher in LVR+A group (Ees: 0.34+/-0.2 versus 0.19+/-0.11 mm Hg/microL). The levels of myocardial ACE activity in LVR+A group was significantly lower than in LVR group. The mRNA expressions of brain natriuretic peptide and transforming growth factor beta1 inducing fibrosis significantly decreased in LV myocardium in LVR+A group. Histologically, myocardial fibrosis was significantly reduced in LVR+A group. CONCLUSIONS: Intravenous administration of ANP had beneficial effects on LV remodeling, function, and fibrosis after LVR. ANP could be a useful intravenous infusion drug for postoperative management after LV repair surgery. PMID- 15364860 TI - Inhibition of matrix metalloproteinase activity by TIMP-1 gene transfer effectively treats ischemic cardiomyopathy. AB - BACKGROUND: Enhanced activity of matrix metalloproteinases (MMPs) has been associated with extracellular matrix degradation and ischemic heart failure in animal models and human patients. This study evaluated the effects of MMP inhibition by gene transfer of TIMP-1 in a rat model of ischemic cardiomyopathy. METHODS AND RESULTS: Rats underwent ligation of the left anterior descending coronary artery with direct intramyocardial injection of replication-deficient adenovirus encoding TIMP-1 (n=8) or null virus as control vector (n=8), and animals were analyzed after 6 weeks. Both systolic and diastolic cardiac function was significantly preserved in the TIMP-1 group compared with control animals (maximum left ventricular [LV] pressure: TIMP-1 70+/-10 versus control 56+/-12 mmHg, P<0.05; maximum dP/dt 2697+/-842 versus 1622+/-527 mmHg/sec, P<0.01; minimum dP/dt -2900+/-917 versus -1195+/-593, P<0.001). Ventricular geometry was significantly preserved in the TIMP-1 group (LV diameter 13.0+/-0.7 versus control 14.4+/-0.4 mm, P<0.001; border-zone wall thickness 1.59+/-0.11 versus control 1.28+/-0.19 mm, P<0.05), and this was associated with a reduction in myocardial fibrosis (2.36+/-0.87 versus control 3.89+/-1.79 microg hydroxyproline/mg tissue, P<0.05). MMP activity was reduced in the TIMP-1 animals (1.5+/-0.9 versus control 43.1+/-14.9 ng of MMP-1 activity, P<0.05). CONCLUSIONS: TIMP-1 gene transfer inhibits MMP activity and preserves cardiac function and geometry in ischemic cardiomyopathy. The reduction in myocardial fibrosis may be primarily responsible for the improved diastolic function in treated animals. TIMP-1 overexpression is a promising therapeutic target for continued investigation. PMID- 15364861 TI - Apelin has in vivo inotropic effects on normal and failing hearts. AB - BACKGROUND: Apelin has been shown ex vivo to be a potent cardiac inotrope. This study was undertaken to evaluate the in vivo effects of apelin on cardiac function in native and ischemic cardiomyopathic rat hearts using a novel combination of a perivascular flow probe and a conductance catheter. METHODS AND RESULTS: Native rats (n =32) and rats in heart failure 6 weeks after left anterior descending coronary artery ligation (n =22) underwent median sternotomy with placement of a perivascular flow probe around the ascending aorta and a pressure volume conductance catheter into the left ventricle. Compared with sham operated rats, the ligated rats had significantly decreased baseline Pmax and max dP/dt. Continuous infusion of apelin at a rate of 0.01 microg/min for 20 minutes significantly increased Pmax and max dP/dt compared with infusion of vehicle alone in both native and failing hearts. Apelin infusion increased cardiac contractility, indicated by a significant increase in stroke volume (SV) without a change in left ventricular end diastolic volume (102+/-16% change from initial SV versus 26+/-20% for native animals, and 110+/-30% versus 26+/-11% for ligated animals), as well as an increase in preload recruitable stroke work (180+/-24 mm Hg versus 107+/-9 mm Hg for native animals). CONCLUSIONS: The present study is the first to show that apelin has positive inotropic effects in vivo in both normal rat hearts and rat hearts in failure after myocardial infarction. Apelin may have use as an acute inotropic agent in patients with ischemic heart failure. PMID- 15364862 TI - Suppression of graft coronary artery disease by a brief treatment with a selective epsilonPKC activator and a deltaPKC inhibitor in murine cardiac allografts. AB - BACKGROUND: Inhibiting delta protein kinase C (deltaPKC) during reperfusion and activating epsilon PKC (epsilonPKC) before ischemia each limits cardiac ischemic injury. Here, we examined whether limiting ischemia-reperfusion injury inhibits graft coronary artery disease (GCAD) and improves murine cardiac allografting. METHODS AND RESULTS: Hearts of FVB mice (H-2q) were transplanted into C57BL/6 mice (H-2b). epsilonPKC activator (psiepsilonRACK) was injected intraperitoneally (20 nmol) into donor mice 20 minutes before procurement. Hearts were then perfused with psiepsilonRACK (1.5 nmol) through the inferior vena cava (IVC) and subsequently submerged in psiepsilonRACK (0.5 micromol/L) for 20 minutes at 4 degrees C. Before reperfusion, the peritoneal cavity of recipients was irrigated with deltaPKC inhibitor (deltaV1-1, 300 nmol); control animals were treated with normal saline. The total ischemic time to the organ was 50 minutes. Two hours after transplantation, production of inflammatory cytokines and adhesion molecules, cardiomyocyte apoptosis, and caspase-3 and caspase-9 (but not caspase 8) activities were significantly reduced in the PKC regulator-treated group. Fas ligand levels (but not Fas) were also significantly reduced in this group. Importantly, GCAD indices, production of inflammatory cytokines, and adhesion molecules were significantly decreased and cardiac allograft function was significantly better as measured up to 30 days after transplantation. CONCLUSIONS: An epsilonPKC activator and a deltaPKC inhibitor together reduced GCAD. Clinically, these PKC isozyme regulators may be useful for organ preservation and prevention of ischemia-reperfusion injury and graft coronary artery disease in cardiac transplantation. PMID- 15364863 TI - Overexpression of human copper/zinc superoxide dismutase (SOD1) suppresses ischemia-reperfusion injury and subsequent development of graft coronary artery disease in murine cardiac grafts. AB - BACKGROUND: Ischemia-reperfusion injury is an important risk factor for graft coronary artery disease (GCAD). We hypothesized that overexpression of SOD1 in donor hearts would suppress ischemia-reperfusion injury and thereby reduce GCAD. METHODS AND RESULTS: In one series, donor hearts of C57BL/6 (H-2b) transgenic mice overexpressing human SOD1 or C57BL/6 wild-type mice were heterotopically transplanted into C57BL/6 recipients and procured after 4 hours of reperfusion (n=6 each). Superoxide, TNF-alpha, and MCP-1/CCL2 production were significantly reduced in the SOD1 transgenic donor heart recipients, and graft injury determined by serum CPK-MB levels was significantly decreased. Cardiomyocyte apoptosis and caspase-3 and caspase-9 activities were significantly decreased in these recipients; caspase-8 activity was unchanged. Fas ligand but not Fas expression was also reduced. In a second series, transgenic and wild-type hearts were transplanted into C-H-2bm12KhEg (H-2bm12) recipients, and then procured on day 56 (n=7 each). Cardiac graft beating was significantly better in the SOD1 transgenic donor heart recipients on days 28, 42, and 56 (but not day 14). Significant reduction in luminal narrowing, the intima/media ratio, and the percentage of diseased vessels was seen in the SOD1 transgenic donor heart recipients, and MCP-1/CCL2, ICAM-1, and VCAM-1 production were significantly reduced. CONCLUSIONS: Overexpression of SOD1 attenuates both apoptosis and the inflammatory response during ischemia-reperfusion injury and therefore mitigates against the subsequent development of GCAD. PMID- 15364864 TI - Cytomegalovirus infection in heart transplant recipients is associated with impaired endothelial function. AB - BACKGROUND: Cardiac allograft vasculopathy (CAV) is initiated by allograft endothelial injury. We hypothesized that a major mechanism by which cytomegalovirus (CMV) could contribute to CAV is by dysregulation of the endothelial vasomotor response. METHODS: Coronary endothelial vasomotor function was determined in 183 consecutive patients (24+/-33 months after transplantation), and was correlated with recipient and donor CMV serological status before transplantation and with documented CMV infection episodes (CMVpp65Ag+). Serial endothelial function measurements were performed in a subgroup of 53 transplant recipients (1 month and 12 months after transplantation). The composite endpoint of cardiovascular related events and death during a follow-up of 66+/-41 months was analyzed based on the CMV serological status before transplantation. RESULTS: The medium event-free time for CMV-negative recipients of CMV-positive hearts was 8.1 years compared with 13.3 years for the other groups (P<0.05). Distal epicardial but not microvascular endothelial function was significantly impaired in CMV seronegative recipients of seropositive donor hearts (n=48) compared with all other groups (P<0.01 versus seronegative recipient/seronegative donor; P<0.05 versus seropositive recipient/seronegative donor; P<0.05 versus seropositive recipient/seropositive donor). Distal epicardial endothelial dysfunction was more pronounced in heart transplant recipients with a history of documented CMV infection compared with patients without any documented CMV infection (P<0.01). In a longitudinal subgroup analysis, distal epicardial and microcirculatory endothelial vasomotor response deteriorated significantly in recipients with documented CMV infection (P<0.05 versus baseline) but not in patients without previous CMV infection. CONCLUSIONS: Documented CMV infection episodes in heart transplant recipients are associated with impaired coronary endothelial function. CMV-negative recipients of CMV-positive donor hearts have an impaired distal epicardial endothelial function and an increased incidence of cardiovascular-related events and death during follow-up. CMV infection may contribute to allograft failure by accelerating coronary endothelial dysfunction. PMID- 15364865 TI - Improved exercise capacity and ischemia 6 and 12 months after transendocardial injection of autologous bone marrow mononuclear cells for ischemic cardiomyopathy. AB - BACKGROUND: We recently reported the safety and feasibility of autologous bone marrow mononuclear cell (ABMMNC) injection into areas of ischemic myocardium in patients with end-stage ischemic cardiomyopathy. The present study evaluated the safety and efficacy of this therapy at 6- and 12-month follow-up. METHODS AND RESULTS: Twenty patients with 6- and 12-month follow-up (11 treated subjects; 9 controls) were enrolled in this prospective, nonrandomized, open-label study. Complete clinical and laboratory evaluations as well as exercise stress (ramp treadmill), 2-dimensional Doppler echocardiography, single-photon emission computed tomography (SPECT) perfusion scanning, and 24-hour Holter monitoring were performed at baseline and follow-up. Transendocardial delivery of ABMMNCs was performed with the aid of electromechanical mapping to identify viable myocardium. Each patient received 15 ABMMNC injections of 0.2 mL each. At 6 and 12 months, total reversible defect, as measured by SPECT perfusion scanning, was significantly reduced in the treatment group as compared with the control group. At 12 months, exercise capacity was significantly improved in the treatment group. This improvement correlated well with monocyte, B-cell, hematopoietic progenitor cell, and early hemapoietic progenitor cell phenotypes. CONCLUSIONS: The 6- and 12-month follow-up data in this study suggest that transendocardial injection of ABMMNCs in patients with end-stage ischemic heart disease may produce a durable therapeutic effect and improve myocardial perfusion and exercise capacity. PMID- 15364866 TI - Role of interleukin-1beta in acute inflammation and graft death after cell transplantation to the heart. AB - BACKGROUND: Poor survival of grafted cells is a major factor hindering the therapeutic effect of cell transplantation; however, the causes of cell death remain unclear. We hypothesized that interleukin-1beta (IL-1beta) might play a role in the acute inflammatory response and graft death after cell transplantation and that inhibition of IL-1beta might improve graft survival. METHODS AND RESULTS: 14C-labeled male skeletal muscle precursor cells were implanted into female mouse hearts by direct intramuscular injection. The amount of 14C-label provides an estimate of the surviving cell number, whereas the amount of male-specific Smcy gene measured by polymerase chain reaction indicates the total (surviving+proliferated) number of donor-derived cells. At 10 minutes after implantation, 44.8+/-2.4% of the grafted cells survived and this steadily decreased to 14.6+/-1.1% by 24 hours, and to 7.9+/-0.6% by 72 hours (n=6 in each point). Proliferation of the surviving cells, which began after 24 hours, resulted in an increase in the total cell number from 15.5+/-0.8% at 24 hours to 24.4+/-1.6% at 72 hours. Acute inflammation was prominent at 24 hours and was reduced by 72 hours, in parallel with IL-1beta expression. Administration of anti IL-1beta antibody improved graft survival at both 24 (25.6+/-1.6%) and 72 hours (14.8+/-1.1%) and resulted in a 2-fold increase in the total cell number at 72 hours (45.8+/-2.4%). The effects of IL-1beta inhibition corresponded with a reduced inflammatory response. CONCLUSIONS: IL-1beta is involved in acute inflammation and graft death after direct intramyocardial cell transplantation. Targeted inhibition of IL-1beta may be a useful strategy to improve graft survival. PMID- 15364867 TI - Targeted cell delivery into infarcted rat hearts by retrograde intracoronary infusion: distribution, dynamics, and influence on cardiac function. AB - BACKGROUND: Intracoronary infusion for cell transplantation has potential advantages in disseminating cells globally into the myocardium with less injury over direct intramuscular injection. Arterial route, however, has a risk of coronary embolism and a limitation in cell delivery into ischemic or infarcted areas. We assessed the efficiency of retrograde intracoronary cell implantation into infarcted hearts using a novel rat model. METHODS AND RESULTS: After left coronary artery ligation in rat, a catheter was inserted into the left cardiac vein, which drains the left ventricular free wall. Through this, 1x10(6) skeletal muscle precursor cells expressing nuclear beta-galactosidase were infused retrogradely into the vein. In situ staining demonstrated that beta-galactosidase expressing donor cells had disseminated throughout the left ventricular free wall, including both infarcted and surrounding border areas, at 10 minutes after infusion. At 28 days, in contrast, positively stained multinuclear myotubes were found in border zones, whereas no positive cells were seen in infarcted areas. Measurement of beta-galactosidase enzyme activity estimated that 29.8+/-6.9% of total infused cells were retained within the myocardium at 10 minutes and that this number decreased to 23.7+/-8.1% at 3 days but rapidly increased thereafter, reaching a plateau at 90.2+/-17.1% by 14 days. Echocardiography and Langendorff perfusion demonstrated that cell implantation improved cardiac function and dimensions by 28 days, compared with both sham-treated and phosphate-buffered saline-infused infarcted hearts, and this was associated with decreased collagen deposition. CONCLUSIONS: Retrograde intracoronary cell transplantation could provide an effective cell delivery into infarcted hearts and could be a useful strategy for treating myocardial infarction. PMID- 15364868 TI - Selective antegrade cerebral perfusion attenuates brain metabolic deficit in aortic arch surgery: a prospective randomized trial. AB - BACKGROUND: Aortic arch surgery has a high incidence of brain injury. This may in part be caused by a cerebral metabolic deficit observed after hypothermic circulatory arrest (HCA). We hypothesized that selective antegrade cerebral perfusion (SACP) would attenuate this phenomenon. METHODS AND RESULTS: In a prospective randomized trial, 42 adult patients were allocated to either HCA (22) or SACP. HCA occurred at a nasopharyngeal temperature of 15 degrees C and SACP at a corporeal temperature of 25 degrees C with cerebral perfusion at 15 degrees C. Paired arterial and jugular venous samples were taken before and after arrest. Continuous transcranial Doppler monitoring of middle cerebral artery velocity (MCAV) was performed. Neuropsychometric testing was performed preoperatively and at 6 and 12 weeks postoperatively. There were 3 hospital deaths (7.1%), 2 strokes (4.8%), and 6 episodes of transient neurological deficit (14.3%). From before to after arrest, jugular bulb pO2 changed by -21.67 mm Hg (26.4) in the HCA group versus +2.27 mm Hg (18.8) in the SACP group (P=0.007). Oxygen extraction changed by +1.7 mL/dL (1.3) in the HCA group versus -1 mL/dL (2.4) in the SACP group (P<0.001). MCAV increased by 6.25 cm/s (9.1) in the HCA group and 19.2 cm/s (10.1) in the SACP group (P=0.001). Incidence of neuropsychometric deficit at 6 weeks was 6/12 (50%) in HCA patients and 8/10 (80%) in SACP patients (P=0.2), and at 12 weeks was 6/16 (38%) in HCA patients and 4/11 (36%) in SACP patients (P=1). CONCLUSIONS: SACP attenuates the metabolic changes seen after HCA. Further studies are required to assess optimal perfusion conditions and clinical outcome. PMID- 15364869 TI - Differences in clinical presentation, management, and outcomes of acute type a aortic dissection in patients with and without previous cardiac surgery. AB - BACKGROUND: There are less data on the clinical and diagnostic imaging characteristics, management, and outcomes of patients with previous cardiac surgery (PCS) presenting with acute type A aortic dissection (AAD). METHODS AND RESULTS: In 617 patients with AAD, we evaluated the differences in the clinical characteristics, management, and in-hospital outcomes of the cohorts with and without PCS. A history of PCS was present in 100 of 617 patients. Patients with PCS were more likely to be males (P=0.02), older (P=0.014), and to have a history of previous aortic dissection (P<0.001) or aneurysms (P<0.001). In contrast, PCS patients were less likely to have presenting chest pain (P<0.001). Cardiac tamponade was less common in PCS patients (P=0.007). Fewer AAD patients with PCS underwent surgical repair (P=0.001). Hospital mortality was not adversely influenced by PCS (odds ratio [OR], 1.46; 95% confidence interval [CI], 0.81 to 2.63), but a trend for increased death was seen in patients with previous aortic valve replacement (AVR) (OR, 2.31; 95% CI, 0.98 to 5.43). Age 70 years or older, previous AVR, shock, and renal failure identified PCS patients at risk for death. CONCLUSIONS: Our study highlights differences in clinical characteristics, management, and outcomes of AAD patients with PCS. Importantly, PCS, with the exception of previous AVR, does not adversely influence early outcomes of AAD patients, including those undergoing surgical repair. However, because of otherwise dismal outcomes with medical management of AAD, our data indicate that a history of PCS (even that of previous AVR) should not preclude physicians from recommending surgical correction of type A aortic dissection in appropriate patients. PMID- 15364870 TI - Evolving strategies for treatment of acute aortic dissection type A. AB - OBJECTIVE: To assess the outcome of 3 different surgical approaches for treatment of acute aortic dissection type A (AADA). METHODS AND RESULTS: Between October 1990 and October 2003, we operated on 295 patients (pts) for AADA. Follow-up was complete for 257 pts (87%). Supracommissural replacement (SCR) of the ascending aorta was applied to 145 pts, 64 pts received a composite replacement (comp), and 48 pts were treated with the aorta valve-sparing (AVS) reimplantation technique. Pts in SCR were older compared with AVS and comp (P=0.002), gender (overall 65% male, P=0.143) and presence of Marfan syndrome (overall 5%, P=0.109) were comparable. Cannulation of the aorta was performed more often in AVS (58%) than in comp (19%) or SCR (22%; P<0.001). Mean operation time, extracorporeal circulation time, and aortic cross-clamp time differ significantly between groups (P<0.001, respectively). Stay in the intensive care unit (P=0.12) and time of hospitalization (P=0.32) were comparable. Overall perioperative mortality was 24% and did not show significant differences between groups (AVS 10.4% versus comp 28% versus SCR 26%; P=0.053). Incidence of neurological complications was similar between groups (P=0.95). Mean time of follow-up was shorter for AVS (19+/-20 months) compared with comp (48+/-48 months) and SCR (46+/-45 months). Survival at 5 years was comparable with 89% for AVS, 85% for comp, and 80% for SCR (P=0.61). Two patients from AVS (4.1%) required reoperation for failure of the reconstructed valve. Pts in comp required less aortic reoperations than pts in SCR (comp 6.3% versus SCR 22%; P=0.005). CONCLUSIONS: In acute aortic dissection type A, the reimplantation technique leads to results comparable to established techniques. Complete removal of diseased tissue, low incidence of reoperation, and lack of anticoagulation may favor this approach in selected patients. PMID- 15364871 TI - Improvement of quality of life after surgery on the thoracic aorta: effect of antegrade cerebral perfusion and short duration of deep hypothermic circulatory arrest. AB - BACKGROUND: We have recently demonstrated that the use of deep hypothermic circulatory arrest (DHCA) during surgery for acute type A aortic dissections or thoracic aortic aneurysms adversely affect mid-term quality of life (QoL). The aim of this study is to assess the impact of DHCA duration and the potential effects of antegrade cerebral perfusion (ACP) on mid-term QoL. METHODS AND RESULTS: Between January 1994 and December 2002, 363 patients underwent surgery of the thoracic aorta with the use of DHCA at our institution. One hundred seventy-six (48.5%) presented with acute type A dissections and 187 (51.5%) presented with aortic aneurysms. ACP was used in 41 (11.3%) cases. All in hospital data were assessed and a follow-up was performed in all survivors after 2.4+/-1.2 years. QoL was analyzed with the Short-Form 36 Health Survey Questionnaire (SF-36). In-hospital mortality was 8.6%. In comparison with patients having undergone DHCA <20 minutes, averaged QoL score was significantly decreased in patients with DHCA between 20 and 34 minutes (95.6+/-12.8 versus 81.9+/-15.7; P<0.01) and >35 minutes (61.8+/-18.3; P<0.01). Averaged QoL score was significantly better with the use of ACP, independently of the duration of DHCA. CONCLUSIONS: DHCA duration >20 minutes, and especially >35 minutes, adversely affects mid-term QoL in patients undergoing surgery of the thoracic aorta. The use of ACP, however, improved averaged QoL score at each time period and allows DHCA to be extended up to 30 minutes, without impairment in mid-term QoL. PMID- 15364872 TI - Growth rate of aortic diameter in patients with type B aortic dissection during the chronic phase. AB - BACKGROUND: The purpose of this study was to evaluate the growth rate of type B double-barrel aortic dissection with computed tomography (CT) and the factors influencing its enlargement. METHODS AND RESULTS: Sixty-two patients were entered into this study, and regular follow-up CT studies (mean; 49.1 months) were performed. The affected aortas and iliac arteries were divided into 5 segments (aortic arch, descending thoracic, suprarenal abdominal, infrarenal abdominal aorta, and iliac artery). Fifty-two of 62 patients (83.9%) had 1 or more segments increased in size during follow-up period. In a total of 177 segments, the presence or absence of blood flow in the false lumen and aortic diameter were evaluated on CT during the follow-up period. The factors (gender, diabetes mellitus, atherosclerotic disease, smoking, entry site in arch, initial diameter, chronic obstructive pulmonary disease, blood pressure, and age) influencing increase in the diameter and growth rate were also evaluated. Of 177 segments, 132 segments (74.6%) increased in size during the follow-up period. The presence of blood flow in the false lumen was the only significant risk factor for increase in the diameter in the univariate and multivariate analysis. The group with blood flow in the false lumen had a significantly higher mean growth rate (3.3 mm/year) than the group without blood flow (-1.4 mm/year) (P<0.0001). The growth rate of aortic dissections in thoracic aorta and abdominal aorta were 4.1 and 1.2 mm/year, respectively. There was a significant difference in the growth rate between the 2 groups (P=0.0003). CONCLUSIONS: In type B aortic dissection, the affected aortas have shown a high incidence of enlargement during the follow up period, and more careful follow-up study is needed for aortic dissections in the thoracic aorta. The presence of blood flow in the false lumen is the most important risk factor for aortic enlargement. PMID- 15364873 TI - Endovascular treatment of thoracic aortic disease: four years of experience. AB - BACKGROUND: The aim of this retrospective study is to investigate efficacy and middle-term results of the stent graft treatment for diseases of descending thoracic aorta. METHODS AND RESULTS: From March 1999 to October 2003, 132 patients (113 male and 19 female, mean age 62+/-14 years) were enrolled. They were divided into 4 groups: aneurysms (43, group A), post-traumatic lesions (24, group B), and complicated type B dissections (43, group C). Twenty-two further patients, with chronic type B dissection and not suitable for endovascular or surgical or hybrid techniques because of multiple entry tears without difference between the true and false lumen and poor clinical conditions, were obliged to receive medical management only (group D). All patients underwent computed tomography (CT) scan and angiography as preoperative assessment. An optimal deployment with exclusion of the aneurysm and/or closure of the entry tear in dissection was achieved in 96.4% (106/110) of the patients that were discharged in good conditions within 6 days. No spinal cord injuries were observed. The follow-up (average 20.82+/-10.01 months, range 1 to 55 months), performed with serial chest CT scans, was 100% complete. No stent graft-related complications were detected, although only in 1 case, an asymptomatic rupture of the Excluder connecting bar was found with a perforation of the fabric and an intra-aortic exposition of the bar itself. In 2 patients with chronic dissection an asymptomatic type II endoleak was detected. A total of 4 hospital deaths resulted in an overall operative mortality of 3.9%. Seven patients (6.3%) died during the follow-up 5 of them for other diseases (4.5%). However, a 40.9% mortality was observed within the obliged medical treatment group. CONCLUSIONS: Endovascular treatment of thoracic aortic diseases, even in the acute phase, may represent a valid option with a low mortality rate. Moreover, the efficacy is proved in the middle-term whereas the long-term follow-up is still pending. PMID- 15364874 TI - Effects of deletion of the tissue inhibitor of matrix metalloproteinases-1 gene on the progression of murine thoracic aortic aneurysms. AB - OBJECTIVE: The cause of thoracic aortic aneurysms (TAAs) is poorly understood. Previous work has suggested an association between development of aortic aneurysms and matrix metalloproteinase (MMP) activity. We hypothesized that removal of the primary endogenous aortic MMP inhibitor (TIMP) through TIMP-1 gene deletion will increase TAA progression. METHODS AND RESULTS: The descending thoracic aortas of wild-type 129 SvE and TIMP-1 gene knockout (TIMP-1-/-) mice were exposed to 0.5 mol/L CaCl2 for 15 minutes, with terminal studies performed at 4 or 8 weeks. TAA lumen diameter was measured using confocal microscopy and normalized to the ascending aorta. In addition, sections were studied with in situ zymography and immunohistochemistry staining for MMP-9. Both wild-type [TAA/ascending ratio (mean+/-SEM): control, 0.85+/-0.02 (n=14); 4 weeks, 1.00+/ 0.03 (n=13); 8 weeks, 1.05+/-0.10 (n=9)] and TIMP-1-/- [control, 0.98+/-0.04 (n=11); 4 weeks, 1.10+/-0.03 (n =21); 8 weeks, 1.22+/-0.09 (n=10)] groups developed aneurysms at 4 and 8 weeks compared with their respective controls (P<0.05). TIMP-1-/- animals developed larger aneurysms than the corresponding wild-type group (P<0.05). Aneurysms in the TIMP-1-/- group were larger at 8 weeks than at 4 weeks (P<0.05), which was not seen in the wild-type aneurysm groups. Both groups showed presence of MMP-9 in 4 and 8 weeks, most prominently in the adventitia and outer media. In situ zymographic activity was increased in the 8 week TIMP-1-/- group compared with wild-type. CONCLUSIONS: Deletion of the TIMP-1 gene results in increased and continued progression of aneurysm formation compared with wild-type mice in a unique TAA model caused at least in part by an alteration in the balance between gelatinase activity and its endogenous inhibition. Therapeutic strategies aimed at modifying MMP activity may reduce or prevent the progression of TAAs. PMID- 15364875 TI - Soluble human complement receptor 1 limits ischemic damage in cardiac surgery patients at high risk requiring cardiopulmonary bypass. AB - BACKGROUND: This study was undertaken to determine whether soluble human complement receptor type 1 (TP10), a potent inhibitor of complement activation, would reduce morbidity and mortality in high-risk patients undergoing cardiac surgery on cardiopulmonary bypass (CPB). METHODS: This was a randomized multicenter, prospective, placebo-controlled, double-blind study in which 564 high-risk patients undergoing cardiac surgery on CPB received an intravenous bolus of TP10 (1, 3, 5, 10 mg/kg) or placebo immediately before CPB. The primary endpoint was the composite events of death, myocardial infarction (MI), prolonged (> or =24 hours) intra-aortic balloon pump support (IABP), and prolonged intubation. RESULTS: TP10 significantly inhibited complement activity after 10 to 15 minutes of CPB and this inhibition persisted for 3 days postoperatively. However, there was no difference in the primary endpoint between the 2 groups (33.7% placebo versus 31.4% TP10; P=0.31). The primary composite endpoint was, however, reduced in all male TP10 patients by 30% (P=0.025). TP10 reduced the incidence of death or MI in males by 36% (P=0.026), the incidence of death or MI in CABG males by 43% (P=0.043) and the need for prolonged IABP support in male CABG and valve patients by 100% (P=0.019). There was, however, no improvement seen in female TP10 patients. There were no significant differences in adverse events between the groups. CONCLUSIONS: TP10 effectively inhibits complement activation during CPB; however, this was not associated with an improvement in the primary endpoint of the study. Nevertheless, TP10 did significantly decrease the incidence of mortality and MI in male patients. PMID- 15364877 TI - Modulation of visceral function by selective stimulation of the left vagus nerve in dogs. AB - The superficial regions of the left vagus nerves of a dog were selectively stimulated with 39-electrode spiral cuffs having 13 circumferential groups of three electrodes (GTE) to modulate the function of the innervated internal organs and glands. Under general anaesthesia, the cuffs were chronically implanted around the nerve in the neck in two adult Beagle dogs and remained viable for 16 months. The regions were stimulated with biphasic, rectangular current pulses (2 mA, 200 micros, 20 Hz) delivered to the group of GTE lying close to the region innervating the specific internal organs or glands. The results showed that specific electrode configurations had actions on the heart (GTE 9), lungs (GTE 4) and pressure in the urinary bladder (GTE 1). It was also shown that GTE no. 10 significantly modified the endocrine function of the pancreas. The results of this study clearly demonstrate that internal organs and glands can be selectively stimulated via the selective stimulation of innervating superficial regions of the autonomous peripheral nerve. PMID- 15364876 TI - Differences in gene expression profiles of diabetic and nondiabetic patients undergoing cardiopulmonary bypass and cardioplegic arrest. AB - BACKGROUND: Diabetes mellitus is an independent risk factor for early postoperative mortality and complications after coronary artery bypass grafting (CABG). We sought to compare the cardiac gene expression responses to cardiopulmonary bypass (CPB) and cardioplegic arrest (C) in patients with and without diabetes. METHODS AND RESULTS: Twenty atrial myocardium samples were harvested from 5 type II insulin-dependent diabetic and 5 matched nondiabetic patients undergoing CABG, before and after CPB/C. Oligonucleotide microarray analyses of 12625 genes were performed on the 10 sample pairs using matched pre CPB tissues as controls. Array results were validated with Northern blotting and immunoblotting. Compared with pre-CPB/C, post-CPB/C myocardial tissues revealed 851 upregulated and 480 downregulated genes with a threshold P< or =0.025 (signal to-noise ratio, 4.04) in the diabetic group, compared with 480 upregulated and 626 downregulated genes (signal-to-noise ratio, 3.04) in the nondiabetic group (P<0.001). There were 18 genes that were upregulated >4-fold in diabetic and nondiabetic patients (including inflammatory/transcription activators FOS, CYR 61, and IL-6, apoptotic gene NR4A1, stress gene DUSP1, and glucose-transporter gene SLC2A3). However, 28 genes showed such marked upregulation in the diabetic group exclusively (including inflammatory/transcription activators MYC, IL8, IL 1beta, growth factor vascular endothelial growth factor, amphiregulin, and glucose metabolism-involved gene insulin receptor substrate 1), and 27 genes in the nondiabetic group only, including glycogen-binding subunit PPP1R3C. CONCLUSIONS: Gene expression profile after CPB/C is quantitatively and qualitatively different in patients with diabetes. These results have important implications for the design of tailored myocardial protection and operative strategies for diabetic patients undergoing CPB/C. PMID- 15364878 TI - Do natriuretic peptides modify arterial baroreflexes in sheep? AB - While atrial and B-type natriuretic peptides (ANP and BNP) have been shown to enhance reflex responses attributed to cardiac vagal afferents, their effects on arterial baroreceptor reflex function remain controversial. The actions of C-type natriuretic peptide (CNP) in this regard are unknown. To clarify their actions on arterial baroreflexes, we tested whether i.v. infusions of ANP, BNP or CNP at 10 pmol kg(-1) min(-1) modified the steady-state mean arterial blood pressure-heart rate (MAP-HR) relationship in conscious sheep. At this dose, all three natriuretic peptides are known to enhance the cardiac chemoreflex response to phenylbiguanide (Bezold-Jarisch reflex). Sigmoid MAP-HR relationships were constructed from the steady-state responses to alternating injections of vasopressor (phenylephrine, 1-15 microg kg(-1)) and vasodepressor agents (nitroprusside, 1-15 microg kg(-1)) in the absence and presence of infused ANP, BNP or CNP (tested in random order at least 1 week apart). No parameter of the steady-state baroreflex relationship was significantly altered by infusion of any of the three natriuretic peptides. We conclude that in conscious sheep, normal arterial baroreceptor-HR reflex function prevails in the presence of moderate doses of ANP, BNP or CNP. PMID- 15364879 TI - Excretion of electrolytes in Brown Norway and Fischer 344 rats: effects of adrenalectomy and of mineralocorticoid and glucocorticoid receptor ligands. AB - Our previous studies showed that adrenalectomy (ADX) has surprisingly no effect on body weight and fluid intake in the Brown Norway rat strain, suggesting that mineralocorticoid receptor (MR)-mediated effects are present even in absence of corticosteroids in this strain. Moreover, glucocorticoid receptor (GR)-mediated mechanisms are more effective in Brown Norway than in Fischer 344 rats. Such functional differences in corticosteroid receptor pathways between Brown Norway and Fischer 344 rats led us to compare the effect of ADX and MR/GR-mediated actions on sodium and potassium excretion between these two rat strains. To this end, we first measured the effect of an acute high dose of aldosterone on the urinary Na+/K+ concentration ratio in intact and ADX Brown Norway and Fischer 344 rats. Second, to discriminate mineralocorticoid from glucocorticoid actions, we treated chronically ADX rats with increasing doses of aldosterone or RU28362, a pure GR agonist, in the drinking fluid. As sodium homeostasis involves salt appetite regulation, behaviour under mineralocorticoid control, we also measured saline preference in Brown Norway and Fischer 344 rats. Our data illustrate: (1) the very limited effect of ADX on body weight, food and fluid intake, diuresis, natriuresis, kaliuresis and salt appetite in Brown Norway rats, supporting the presence of MR signalling pathways in the absence of adrenal steroids in these rats; (2) the insensitivity of MR to aldosterone in intact Brown Norway rats, and the reduced sensitivity of MR to aldosterone in ADX Brown Norway rats compared with ADX Fischer 344 rats; and (3) the greater sensitivity of GR-related mechanisms to RU28362 in Brown Norway than in Fischer 344 rats in terms of body weight gain and electrolyte excretion. Considering that both MRs and GRs regulate hypothalamic-pituitary-adrenal axis processes, such functional differences in corticosteroid receptors could be at the origin, at least partly, of the strain differences in corticotropic activity/reactivity to stress previously described. PMID- 15364880 TI - Muscle afferent contributions to the cardiovascular response to isometric exercise. AB - The cardiovascular response to isometric exercise is governed by both central and peripheral mechanisms. Both metabolic and mechanical stresses on the exercising skeletal muscle produce cardiovascular change, yet it is often overlooked that the afferent signal arising from the muscle can be modified by factors other than exercise intensity. This review discusses research revealing that muscle fibre type, muscle mass and training status are important factors in modifying this peripheral feedback from the active muscles. Studies in both animals and humans have shown that the pressor response resulting from exercise of muscle with a faster contractile character and isomyosin content is greater than that from a muscle of slower contractile character. Athletic groups participating in training programmes that place a high anaerobic load on skeletal muscle groups show attenuated muscle afferent feedback. Similarly, longitudinal studies have shown that specific local muscle training also blunts the pressor response to isometric exercise. Thus it appears that training may decrease the metabolic stimulation of muscle afferents and in some instances chronic exposure to the products of anaerobic metabolism may blunt the sensitivity of the muscle metaboreflex. There may be surprising parallels between the local muscle conditions induced in athletes training for longer sprint events (e.g. 400 m) and by the low-flow conditions in, for example, the muscles of chronic heart failure patients. Whether their similar attenuations in muscle afferent feedback during exercise are due to decreased metabolite accumulation or to a desensitization of the muscle afferents is not yet known. PMID- 15364881 TI - Hypo-osmotic potentiation of acetylcholine-stimulated ciliary beat frequency through ATP release in rat tracheal ciliary cells. AB - The ciliary beat frequency (CBF) of rat tracheal ciliary cells in a slice preparation was measured using video-enhanced contrast (VEC) microscopy. Acetylcholine (ACh) increased CBF mediated via intracellular Ca2+ concentration ([Ca2+]i) in a dose-dependent manner. An adequate hypo-osmotic stress (-40 mosM) potentiated ACh-stimulated CBF increase in tracheal ciliary cells and shifted the ACh dose-response curve to the left (lower concentration side). This potentiation was independent of hypo-osmotic stresses applied ranging from -20 mosM to -90 mosM. A hypo-osmotic stress induces ATP release in many cell types. The present study demonstrated that suramin (an inhibitor of purinergic receptors) and apyrase (an ATPase/ADPase) eliminate the hypo-osmotic potentiation of ACh stimulated CBF increase and that ATP increased [Ca2+]i and CBF, as well as potentiating ACh-stimulated rises in [Ca2+]i and CBF increase. Moreover, the apical surface of tracheal ciliary cells were stained immunopositive for the P2X4 purinergic receptor. A hypo-osmotic stress (-40 mosM) transiently increased [Ca2+]i and potentiated the ACh-stimulated [Ca2+]i increase. The hypo-osmotic potentiation of ACh-stimulated CBF increase was not detected under Ca2+-free conditions. These observations suggest that a hypo-osmotic stress stimulates ATP release from the trachea. The released ATP may induce further increases in [Ca2+]i and CBF in ACh-stimulated tracheal ciliary cells, which may be mediated by purinergic receptors, such as P2X4. PMID- 15364882 TI - Uncoupling of rhythmic hypoglossal from phrenic activity in the rat. AB - During eupnoea, rhythmic motor activities of the hypoglossal, vagal and phrenic nerves are linked temporally. The inspiratory discharges of the hypoglossal and vagus motor neurones commence before the onset of the phrenic burst. The vagus nerve also discharges in expiration. Upon exposure to hypocapnia or hypothermia, the hypoglossal discharge became uncoupled from that of the phrenic nerve. This uncoupling was evidenced by variable times of onset of hypoglossal discharge before or after the onset of phrenic discharge, extra bursts of hypoglossal activity in neural expiration, or complete absence of any hypoglossal discharge during a respiratory cycle. No such changes were found for vagal discharge, which remained linked to the phrenic bursts. Intracellular recordings in the hypoglossal nucleus revealed that all changes in hypoglossal discharge were due to neuronal depolarization. These results add support to the conclusion that the brainstem control of respiratory-modulated hypoglossal activity differs from control of phrenic and vagal activity. These findings have implications for any studies in which activity of the hypoglossal nerve is used as the sole index of neural inspiration. Indeed, our results establish that hypoglossal discharge alone is an equivocal index of the pattern of overall ventilatory activity and that this is accentuated by hypercapnia and hypothermia. PMID- 15364883 TI - Survivin is an independent prognostic marker for risk stratification of breast cancer patients. AB - BACKGROUND: Results in previous qualitative studies of the association of the apoptosis inhibitor survivin with prognosis of breast cancer patients have been contradictory. METHODS: Survivin mRNA was measured by quantitative TaqMan reverse transcription-PCR in 275 breast cancer tissues from patients with operable tumors and was correlated with established clinicopathologic factors, relapse-free survival [(RFS); 102 events], and overall survival [(OS); 81 events]. RESULTS: High survivin mRNA concentrations were found mainly in tissues from younger patients and in high-grade cancer tissues. High survivin concentrations were most strongly associated with estrogen receptor- or progesterone receptor-negative tumors. In univariate Cox regression analysis for RFS, survivin concentrations were significantly associated with poor prognosis with a hazard ratio (HR) of 1.99 (95% confidence interval, 1.31-3.02; P = 0.001) for every 10-fold increase in expression. For OS, a significant contribution of survivin to poor prognosis was found with a HR of 2.76 (1.67-4.55; P <0.001). Multivariate analyses were performed including established clinicopathologic factors. For RFS, age (P = 0.027), nodal category (P <0.001), and survivin [HR = 1.78 (1.18-2.68); P = 0.006] contributed significantly to the model. For OS, only nodal category (P <0.001) and survivin [HR = 3.05 (1.83-5.10); P <0.001] were significant. CONCLUSION: Survivin demonstrates a strong, independent, association with poor prognosis. Survivin might be used as a new marker to stratify breast cancer patients for more optimal treatment modalities, or it could be a promising new target for therapy. PMID- 15364885 TI - Evaluation of quality-control criteria for microarray gene expression analysis. AB - BACKGROUND: Development of quality-control criteria to ensure reproducibility of microarray results for potential clinical application is still in its infancy. METHODS: In the present studies we developed quality-control criteria and evaluated their effect in microarray data analysis using total RNA from cell lines, frozen tumors, and a commercially available reference RNA. Quality-control criteria such as A(260)/A(280) ratios, percentage of rRNA, and median size of cDNA and cRNA synthesis products were evaluated for robustness in microarray analysis. Furthermore, precision studies using a reference material were performed on the Affymetrix HG-U133A high-density oligonucleotide microarrays. The same reference RNA sample was examined in 16 different chips run on 2 different days in the four different modules of the Affymetrix fluidics workstation. Fresh and frozen fragmented cRNAs were also compared. An ANOVA model was fit to identify the main sources of variation. RESULTS: Good-quality samples showed >30% rRNA in the electropherograms and cDNA and cRNA synthesis products with median sizes of 2.0 and 3.0 kb, respectively. Precision studies showed that the main source of variation was the day-to-day variability, minimally affecting hybridization exogenous control genes. Altogether, the results showed that the Affymetrix Genechip system is highly reproducible when RNA that meet the quality control criteria are used (overall P >0.01). CONCLUSIONS: These results confirm the need to establish defined quality-control criteria for sample quality to distinguish between analytical and biological variability. PMID- 15364884 TI - Protein chip array profiling analysis in patients with severe acute respiratory syndrome identified serum amyloid a protein as a biomarker potentially useful in monitoring the extent of pneumonia. AB - BACKGROUND: A new strain of coronavirus (CoV) has caused an outbreak of severe acute respiratory syndrome (SARS), with 8098 individuals being infected and 774 deaths worldwide. We carried out protein chip array profiling analysis in an attempt to identify biomarkers that might be useful in monitoring the clinical course of SARS patients. METHODS: We performed surface-enhanced laser desorption ionization time-of-flight mass spectrometry on 89 sera collected from 28 SARS patients, 72 sera from 51 control patients with various viral or bacterial infections, and 10 sera from apparently healthy individuals. RESULTS: Nine significantly increased and three significantly decreased serum biomarkers were discovered in the SARS patients compared with the controls. Among these biomarkers, one (11,695 Da) was identified to be serum amyloid A (SAA) protein by peptide mapping and tandem mass spectrometric analysis. When we monitored the SAA concentrations longitudinally in 45 sera from four SARS patients, we found a good correlation of SAA concentration with the extent of pneumonia as assessed by a serial chest x-ray opacity score. Increased SAA occurred in three of four patients at the time of extensive pneumonia as indicated by high x-ray scores. Over the course of gradual recovery in two patients, as assessed clinically and radiologically, SAA concentrations gradually decreased. In the third patient, the concentrations were initially increased, but were further increased with superimposed multiple bacterial infections. SAA was not markedly increased in the fourth patient, who had low x-ray scores and whose clinical course was relatively mild. CONCLUSIONS: Protein chip array profiling analysis could be potentially useful in monitoring the severity of disease in SARS patients. PMID- 15364886 TI - High-sensitivity detection of the A3243G mutation of mitochondrial DNA by a combination of allele-specific PCR and peptide nucleic acid-directed PCR clamping. AB - BACKGROUND: The A3243G mutation of mitochondrial DNA (mtDNA) is involved in many common diseases, including diabetes mellitus and mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS). For detection of this mutation, allele-specific PCR is highly sensitive but requires strict control of PCR conditions; it thus is not adequate for a routine clinical test. We aimed to develop a routinely available PCR method for quantitative detection of low-level heteroplasmy of the A3243G mutation. METHODS: Quantitative allele-specific PCR for the A3243G mutation was performed in the presence of peptide nucleic acid (PNA), in which PNA is complementary to the wild-type mtDNA, with one primer having a 3' end matched to nucleotide position 3243 of the mutant. RESULTS: With our method, amplification of wild-type mtDNA was suppressed 7000-fold compared with amplification of the mutant mtDNA under a broad range of conditions: DNA, 5 100 ng; annealing temperature, 61-66 degrees C; and PNA, 1.5-3.5 micromol/L. Hence, 0.1% heteroplasmy of the A3243G mutation can be reliably quantified by this method. Blood samples form 40 healthy volunteers showed <0.06% heteroplasmy, suggesting that 0.1% is diagnostically significant. CONCLUSIONS: PNA maintains the specificity of allele-specific PCR over a wide range of conditions, which is important for routine clinical testing. PMID- 15364887 TI - Analytical validation of the tag-it high-throughput microsphere-based universal array genotyping platform: application to the multiplex detection of a panel of thrombophilia-associated single-nucleotide polymorphisms. AB - BACKGROUND: We have developed a novel, microsphere-based universal array platform referred to as the Tag-It platform. This platform is suitable for high-throughput clinical genotyping applications and was used for multiplex analysis of a panel of thrombophilia-associated single-nucleotide polymorphisms (SNPs). METHODS: Genomic DNA from 132 patients was amplified by multiplex PCR using 6 primer sets, followed by multiplex allele-specific primer extension using 12 universally tagged genotyping primers. The products were then sorted on the Tag-It array and detected by use of the Luminex xMAP system. Genotypes were also determined by sequencing. RESULTS: Empirical validation of the universal array showed that the highest nonspecific signal was 3.7% of the specific signal. Patient genotypes showed 100% concordance with direct DNA sequencing data for 736 SNP determinations. CONCLUSIONS: The Tag-It microsphere-based universal array platform is a highly accurate, multiplexed, high-throughput SNP-detection platform. PMID- 15364888 TI - Multi-biomarker risk stratification of N-terminal pro-B-type natriuretic peptide, high-sensitivity C-reactive protein, and cardiac troponin T and I in end-stage renal disease for all-cause death. AB - BACKGROUND: In patients with end-stage renal disease (ESRD), the ability of single and multiple biomarker monitoring to predict adverse outcomes has not been well established. This study determined the prognostic value of multiple biomarkers for all-cause death over 2 years in 399 ESRD patients. METHODS: The risk of all-cause death was determined by use of multiple biomarkers based on concentrations for a reference population (normal) and cutoffs based on tertile distributions in the ESRD group. Biomarkers studied included N-terminal pro-B type natriuretic peptide (NT-proBNP), high-sensitivity C-reactive protein (hsCRP; Dade Behring and Roche assays), and cardiac troponin T (cTnT; Roche) and I (cTnI; Dade Behring and Beckman Coulter assays). Relative risks of death were estimated and survival curves computed. RESULTS: A total of 101 deaths occurred during 594 patient-years of follow-up. Increased NT-proBNP concentrations were not predictive of death on the basis of the normal cutoffs. However, tertile analysis of NT-proBNP was significantly predictive of death and had a ROC area under the curve equivalent to or better than any of the other biomarkers. Biomarkers independently predictive of survival were hsCRP (P <0.001, either assay), cTnT (P <0.05), and cTnI (Dade, P <0.05). Two-year mortality rates were 6% (n = 45) with normal hsCRP, cTnI, and cTnT concentrations; 19% (n = 173) with increased hsCRP or cTnT and normal cTnI; 44% (n = 160) with both hsCRP and cTnT increased and normal cTnI; 61% (n = 21) with increased cTnI (Dade) or 47% (n = 74) with increased cTnI (Beckman) regardless of hsCRP or cTnT concentrations. Defined by the normal cutoffs, increased concentrations of biomarkers were present in various proportions of the 399 patients with ESRD: NT-proBNP, 99%; hsCRP, 46% (both Roche and Dade assays); cTnT, 85%; cTnI, 19% (Beckman assay) and 5% (Dade assay). CONCLUSIONS: Although mechanisms likely vary for causation, increased plasma hsCRP, cTnT, and cTnI above the cutoffs for our reference (normal) population were all independently predictive of subsequent death in ESRD patients. Tertile analysis for NT-proBNP also demonstrated prognostic value. PMID- 15364889 TI - Reverse transcription-multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Vibrio cholerae O1, and Salmonella Typhi. AB - BACKGROUND: Escherichia coli O157:H7, Vibrio cholerae O1, and Salmonella Typhi are pathogenic bacteria that can be found in contaminated water supplies throughout the world. No currently available assays can simultaneously detect and identify all three pathogens. Our aim was to develop a rapid and reliable technique for simultaneous detection of these pathogens. METHODS: Four unique genes were chosen as the targets of detection. Forward and reverse primers were designed to specifically amplify different sizes of these target genes: a 239-bp region of the E. coli O157 lipopolysaccharide (LPS) gene (rfbE); a 179-bp region of the H7 flagellin gene (fliC); a 419-bp region of the V. cholerae O1 LPS gene (rfbE); and a 329-bp region of Salmonella Typhi LPS gene (tyv). To ensure the detection of only viable replicating bacteria, RNA was extracted for analysis. After reverse transcription, cDNAs were simultaneously amplified in a single tube by multiplex PCR. The multiplex PCR products were analyzed by gel electrophoresis. To characterize the assay we analyzed, in a blinded fashion, seven unknown RNA samples containing various combinations of total RNA from these bacteria as well as clinical isolates. RESULTS: All seven unknown RNA samples were correctly identified. The assay was able to detect and identify as few as 30 cells of E. coli O157:H7 and Salmonella Typhi in clinical isolates, and the presence of other bacteria did not interfere with the analysis. CONCLUSION: An assay combining reverse transcription with single-tube multiplex PCR was successfully developed and validated for simultaneous detection of viable E. coli O157:H7, V. cholerae O1, and Salmonella Typhi. PMID- 15364890 TI - Platelet-activating factor acetylhydrolase is not associated with carotid intima media thickness in hypercholesterolemic Sicilian individuals. AB - BACKGROUND: Atherosclerosis is a complex, chronic disease that usually arises from the converging action of several pathogenic processes, including hypertension, hyperlipidemia, obesity, and the accumulation of oxidized LDL. Platelet-activating factor acetylhydrolase (PAF-AH) is a LDL- and HDL-bound enzyme that hydrolyzes and inactivates PAF and prevents LDL-cholesterol oxidation, thus delaying the onset of atherosclerotic disease. METHODS: We evaluated the relationship between variants of the PAF-AH gene polymorphisms Arg92His, Ile198Thr, and Ala379Val and the presence of carotid atherosclerosis in 190 hypercholesterolemic Sicilian individuals. Carotid artery intima-media wall thickness (IMT) was measured as an indicator of early atherosclerotic disease. The participants were classified according to having normal (< or =1 mm) or abnormal (> or =1 mm) IMT and were also investigated for physical characteristics and biochemical indices, including PAF-AH activity. RESULTS: PAF-AH activity and LDL concentrations were significantly correlated in hypercholesterolemic patients, but plasma PAF-AH activity and HDL were not significantly correlated in either IMT group. No significant differences were detected among the PAF-AH gene polymorphisms in both groups after correction for age, sex, body mass index, plasma glucose and lipid concentrations, PAF-AH activity, blood pressure, and smoking habits. The analysis of PAF-AH genotype distribution showed no significant differences in percentage of 92, 198, and 379 genotypes in both IMT groups. CONCLUSION: Our data provided no evidence that PAF-AH polymorphisms influence PAF-AH activity and atherosclerosis in hypercholesterolemic Sicilian patients. PMID- 15364891 TI - Genetic predisposition of the interleukin-6 response to inflammation: implications for a variety of major diseases? AB - BACKGROUND: A single-nucleotide polymorphism (SNP) in the promoter region of the interleukin-6 (IL-6) gene at position -174 (G>C) has been reported to be associated with a variety of major diseases, such as Alzheimer disease, atherosclerosis, and cardiovascular disease, cancer, non-insulin-dependent diabetes mellitus, osteoporosis, sepsis, and systemic-onset juvenile chronic arthritis. However, authors of previous in vitro and in vivo studies have reported conflicting results regarding the functionality of this polymorphism. We therefore aimed to clarify the role of the -174 SNP for the induction of IL-6 in vivo. METHODS: We vaccinated 20 and 18 healthy individuals homozygous for the 174 C and G alleles, respectively, with 1 mL of Salmonella typhii vaccine. IL 1beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha) were measured in the blood at baseline and up to 24 h after vaccination. RESULTS: Individuals with the G genotype had significantly higher plasma IL-6 values at 6, 8, and 10 h after vaccination than did individuals with the C genotype (P <0.005). There were no differences between the two genotypes regarding serum concentrations of IL-1beta and TNF-alpha before or after vaccination. CONCLUSIONS: The -174 G>C SNP in the promoter region of the IL-6 gene is functional in vivo with an increased inflammatory response associated with the G allele. Considering the central role of IL-6 in a variety of major diseases, the present finding might be of major relevance. PMID- 15364892 TI - Immunoquantification of alpha-galactosidase: evaluation for the diagnosis of Fabry disease. AB - BACKGROUND: Fabry disease is an X-linked inborn error of glycosphingolipid catabolism resulting from a deficiency of the lysosomal exoglycohydrolase, alpha galactosidase. Enzyme replacement therapy is currently available for Fabry disease, but early diagnosis before the onset of irreversible pathology will be mandatory for successful treatment. Presymptomatic detection would be possible through the use of a newborn-screening program. We report on the use of sensitive assays for the measurement of alpha-galactosidase protein and activity and for the protein saposin C, which are diagnostic markers for Fabry disease. METHODS: Two sensitive immunoassays for the measurement of alpha-galactosidase activity and protein were used to determine the concentrations of alpha-galactosidase in dried filter-paper blood spots and plasma samples from control patients and patients with a lysosomal storage disorder (LSD). RESULTS: Fabry hemizygous individuals were clearly identified from control populations by decreases in both alpha-galactosidase activity and protein. Fabry heterozygotes generally fell between the hemizygotes and controls. Including the measurement of saposin C enabled differentiation between Fabry heterozygotes and controls. In blood spots, all Fabry individuals could be distinguished from control blood spots as well as from 16 other LSD patients. CONCLUSIONS: The determination of alpha-galactosidase activity or protein in dried filter-paper blood spots could be used for the diagnosis of Fabry patients. With further validation, these assays could be used for the identification of Fabry patients in newborn-screening programs and may also be suitable for screening high-risk populations. PMID- 15364893 TI - Non-immunostimulatory nonviral vectors. AB - The vectors for gene delivery are usually classified as viral and nonviral vectors. While the viral vectors are very efficient in transducing cells, safety concerns regarding their use in humans make nonviral vectors an attractive alternative. Among the nonviral vectors, the lipoplexes (complexes of cationic liposome/pDNA) are the most studied and represent the most promising approaches for human clinical trials. However, an inflammatory response is invariably associated with administration of the lipoplexes, which must be avoided in the clinical application. Here, we have successfully developed a nonimmunostimulatory vector for gene therapy. The vector possesses dual functions of: 1) efficiently delivering a gene to target cells and 2) codelivering DNA and inflammatory suppressors into the immune cells where the released suppressor can inhibit cytokine production. The inflammatory suppressors successfully delivered by the vector included glucocorticoids, a nonsteroidal anti-inflammatory drug (NSAID), an NF-kappaB inhibitor, and a natural compound from an herbal medicine. Intravenous injection of the vector dramatically suppressed the cytokine production induced by CpG motif pDNA, including TNF-alpha, IL-12 and IFN-gamma. This new gene vector has a great potential in clinical gene therapy. Another potential use of the vector is codelivery of an enhancer candidate, acting at the transcriptional and translational levels to improve the efficiency of gene transfer by the nonviral vector. Moreover, the unique feature of this vector is that it can be used as an easy and powerful tool for in vivo screening of anti inflammatory drugs. PMID- 15364894 TI - Increased arginase II and decreased NO synthesis in endothelial cells of patients with pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH), a fatal disease of unknown etiology characterized by impaired regulation of pulmonary hemodynamics and vascular growth, is associated with low levels of pulmonary nitric oxide (NO). Based upon its critical role in mediating vasodilation and cell growth, decrease of NO has been implicated in the pathogenesis of PAH. We evaluated mechanisms for low NO and pulmonary hypertension, including NO synthases (NOS) and factors regulating NOS activity, i.e. the substrate arginine, arginase expression and activity, and endogenous inhibitors of NOS in patients with PAH and healthy controls. PAH lungs had normal NOS I-III expression, but substrate arginine levels were inversely related to pulmonary artery pressures. Activity of arginase, an enzyme that regulates NO biosynthesis through effects on arginine, was higher in PAH serum than in controls, with high-level arginase expression localized by immunostaining to pulmonary endothelial cells. Further, pulmonary artery endothelial cells derived from PAH lung had higher arginase II expression and produced lower NO than control cells in vitro. Thus, substrate availability affects NOS activity and vasodilation, implicating arginase II and alterations in arginine metabolic pathways in the pathophysiology of PAH. PMID- 15364895 TI - Ilf3 and NF90 associate with the axonal targeting element of Tau mRNA. AB - In neurons, the selective translocation of Tau mRNA toward axons is due to the presence of a nucleotide sequence located in its 3' untranslated region and serving as axonal targeting element. Using this RNA sequence as a probe by a Northwestern approach, we have detected several proteins that interact with the targeting RNA element and could potentially be involved in Tau mRNA translocation, translation halting, and/or stabilization. Among them, two proteins were identified as the interleukin enhancer binding factor 3 (Ilf3) and NF90, two isoforms derived from a single gene product through alternative splicing. Each protein comprises two double-stranded RNA binding motifs that can interact with the predicted stem-loop secondary structure of the axonal targeting element. Specific antibodies raised against common or specific peptide sequences showed that both Ilf3 and NF90 are polymorphic proteins that are detected in neuronal nuclei and cell bodies, as well as in the proximal neuritic segments. This observation favors the idea that Ilf3 and NF90 are part of a protein complex that escorts Tau mRNA toward the axon. PMID- 15364896 TI - Amino-truncated amyloid beta-peptide (Abeta5-40/42) produced from caspase-cleaved amyloid precursor protein is deposited in Alzheimer's disease brain. AB - Caspase activation and apoptosis are implicated in Alzheimer's disease (AD). In view of the finding that the amyloid precursor protein (APP) undergoes caspase mediated cleavage in the cytoplasmic region, we analyzed amyloid beta-peptide (Abeta) production in human neuronal and nonneuronal cells expressing wild-type APP and the caspase-cleaved form of APP (APPDeltaC). Biochemical analyses, including immunoprecipitation/mass spectrometry, revealed that APPDeltaC expressing cells secrete increased levels of amino-terminally truncated Abeta5 40/42 and reduced levels of Abeta1-40/42, compared with wild-type APP-expressing cells. We propose that Abeta5-40/42 is derived from alternative beta-cleavage of APP by alpha-secretase-like protease(s), based on data from treatment of cells with inhibitors of BACE and alpha-secretase. Apoptosis induction resulted in this alternative cleavage of APP in wild-type APP-expressing cells. Moreover, immunohistochemical staining of the AD brain with an end-specific antibody to Abeta5-40/42 revealed peptide deposits in vascular lesions with amyloid angiopathy. The data collectively suggest that caspase cleavage of APP leads to increased production and deposition of Abeta5-40/42 in the AD brain, and highlight the significance of amino-truncated Abeta in the pathogenesis of AD. PMID- 15364897 TI - Endothelial NO synthase genotype and risk of preeclampsia: a multicenter case control study. AB - Polymorphisms in the endothelial NO synthase (eNOS) gene have been evaluated as risk factors for preeclampsia. However, data from small studies are conflicting. We assessed whether eNOS genotypes alter the risk of preeclampsia in a population in which the incidence of this disorder is high. A total of 844 young pregnant women (322 preeclamptic and 522 controls) were recruited from 5 cities. Genotyping for the Glu298Asp, intron-4 and -786T-->C polymorphisms in the eNOS gene was conducted. Multivariate odds ratios (ORs) were obtained to estimate the association of individual polymorphisms and haplotypes with preeclampsia risk. No increase in the risk of preeclampsia for the intron-4 or -786T-->C polymorphisms was observed under any model of inheritance. In contrast, in women homozygous for the Asp298 allele, the adjusted OR for preeclampsia was 4.60 (95% confidence interval [CI], 1.73 to 12.22) compared with carriers of the Glu298 allele. After a multivariate analysis, carriage of the "Asp298-786C-4b" haplotype was also associated with increased risk of preeclampsia (OR, 2.11 [95% CI, 1.33 to 3.34]) compared with carriers of the "Glu298-786T-4b" haplotype. The eNOS Glu298Asp polymorphism and the Asp298-786C-4b haplotype are risk factors for preeclampsia. PMID- 15364898 TI - Epistatic interaction between beta2-adrenergic receptor and neuropeptide Y genes influences LDL-cholesterol in hypertension. AB - Beta2-adrenergic receptor gene and neuropeptide Y gene may potentially influence lipid metabolism and overall energy balance. Therefore, we examined associations of these genes with lipid fractions and obesity-related phenotypes in hypertensive subjects. A total of 638 white individuals from 212 Polish families with clustering of essential hypertension were phenotyped for cardiovascular risk determinants. Each subject was genotyped for functional polymorphisms of beta2 adrenergic receptor gene (Arg16Gly and Gln27Glu) and neuropeptide Y (Leu7Pro). Of 3 common haplotypes of beta2-adrenergic receptor gene, Arg16Gln27 was overtransmitted to offspring with elevated levels of total cholesterol (Z=2.2; P=0.026) and LDL-cholesterol (Z=3.2; P=0.002). Individually, Leu7Pro was not associated with any of the metabolic phenotypes in family-based tests or case control analyses. However, in the presence of Arg allele of Arg16Gly and Gln allele of Gln27Glu, homozygosity for Leu variant of the Leu7Pro polymorphism was associated with 2.1-increased odds ratio (confidence interval, 1.10 to 3.81; P=0.024) of elevated LDL in hypertensive subjects, independent of age, gender, body mass index, adjusted blood pressures, antihypertensive therapy, and use of nonselective beta-blockers and diuretics. Consistently, there was a significant multilocus association among variants of Arg16Gly, Gln27Glu, and Leu7Pro in hypertensive probands with elevated LDL (cases; P=0.028) but not in hypertensive subjects with normal LDL (controls). This study revealed an association of LDL cholesterol with beta2-adrenergic receptor gene haplotype and provided evidence for epistatic interaction between beta2-adrenergic receptor gene and neuropeptide Y gene in determination of LDL-cholesterol in patients with essential hypertension. PMID- 15364899 TI - Importance of achieving lower blood pressure in hypertensive patients with diabetes. PMID- 15364900 TI - Pattern of sequence variation across 213 environmental response genes. AB - To promote the clinical and epidemiological studies that improve our understanding of human genetic susceptibility to environmental exposure, the Environmental Genome Project (EGP) has scanned 213 environmental response genes involved in DNA repair, cell cycle regulation, apoptosis, and metabolism for single nucleotide polymorphisms (SNPs). Many of these genes have been implicated by loss-of-function mutations associated with severe diseases attributable to decreased protection of genomic integrity. Therefore, the hypothesis for these studies is that individuals with functionally significant polymorphisms within these genes may be particularly susceptible to genotoxic environmental agents. On average, 20.4 kb of baseline genomic sequence or 86% of each gene, including a substantial amount of introns, all exons, and 1.3 kb upstream and downstream, were scanned for variations in the 90 samples of the Polymorphism Discovery Resource panel. The average nucleotide diversity across the 4.2 MB of these 213 genes is 6.7 x 10(-4), or one SNP every 1500 bp, when two random chromosomes are compared. The average candidate environmental response gene contains 26 PHASE inferred haplotypes, 34 common SNPs, 6.2 coding SNPs (cSNPs), and 2.5 nonsynonymous cSNPs. SIFT and Polyphen analysis of 541 nonsynonymous cSNPs identified 57 potentially deleterious SNPs. An additional eight polymorphisms predict altered protein translation. Because these genes represent 1% of all known human genes, extrapolation from these data predicts the total genomic set of cSNPs, nonsynonymous cSNPs, and potentially deleterious nonsynonymous cSNPs. The implications for the use of these data in direct and indirect association studies of environmentally induced diseases are discussed. PMID- 15364901 TI - Identification of mammalian microRNA host genes and transcription units. AB - To derive a global perspective on the transcription of microRNAs (miRNAs) in mammals, we annotated the genomic position and context of this class of noncoding RNAs (ncRNAs) in the human and mouse genomes. Of the 232 known mammalian miRNAs, we found that 161 overlap with 123 defined transcription units (TUs). We identified miRNAs within introns of 90 protein-coding genes with a broad spectrum of molecular functions, and in both introns and exons of 66 mRNA-like noncoding RNAs (mlncRNAs). In addition, novel families of miRNAs based on host gene identity were identified. The transcription patterns of all miRNA host genes were curated from a variety of sources illustrating spatial, temporal, and physiological regulation of miRNA expression. These findings strongly suggest that miRNAs are transcribed in parallel with their host transcripts, and that the two different transcription classes of miRNAs ('exonic' and 'intronic') identified here may require slightly different mechanisms of biogenesis. PMID- 15364902 TI - A novel class of mammalian-specific tailless retropseudogenes. AB - In addition to their central function in protein biosynthesis, tRNAs also play a pervasive role in genome evolution and architecture because of their extensive ability to serve as templates for retroposition. Close to half of the human genome consists of discernible transposable elements, a vast majority of which are derived from RNA via reverse transcription and genomic integration. Apart from the presence of direct repeats (DRs) that flank the integrated sequence of retroposons, genomic integrations are usually marked by an oligo(A) tail. Here, we describe a novel class of retroposons that lack A-tails and are therefore termed tailless retropseudogenes. Analysis of approximately 2500 tRNA-related young tailless retropseudogene sequences revealed that they comprise processed and unprocessed (pre-)tRNAs, 3'-truncated in their loop regions, or truncated tRNA-derived SINE RNAs. Surprisingly, their mostly nonrandom integration is dependent on the priming of reverse transcription at sites determined by their 3' terminal 2-18 nucleotides and completely independent from oligoadenylation of the template RNA. Thus, tailless retropseudogenes point to a novel, variant mechanism for the biogenesis of retrosequences. PMID- 15364903 TI - Human, mouse, and rat genome large-scale rearrangements: stability versus speciation. AB - Using paired-end sequences from bacterial artificial chromosomes, we have constructed high-resolution synteny and rearrangement breakpoint maps among human, mouse, and rat genomes. Among the >300 syntenic blocks identified are segments of over 40 Mb without any detected interspecies rearrangements, as well as regions with frequently broken synteny and extensive rearrangements. As closely related species, mouse and rat share the majority of the breakpoints and often have the same types of rearrangements when compared with the human genome. However, the breakpoints not shared between them indicate that mouse rearrangements are more often interchromosomal, whereas intrachromosomal rearrangements are more prominent in rat. Centromeres may have played a significant role in reorganizing a number of chromosomes in all three species. The comparison of the three species indicates that genome rearrangements follow a path that accommodates a delicate balance between maintaining a basic structure underlying all mammalian species and permitting variations that are necessary for speciation. PMID- 15364905 TI - Acute effects of parainfluenza virus on epithelial electrolyte transport. AB - Parainfluenza viruses are important causes of respiratory disease in both children and adults. In particular, they are the major cause of the serious childhood illness croup (laryngotracheobronchitis). The infections produced by parainfluenza viruses are associated with the accumulation of ions and fluid in the respiratory tract. It is not known, however, whether this accumulation is because of a direct effect of the viruses on ion and fluid transport by the respiratory epithelium. Here we show that a model parainfluenza virus (the Sendai virus), in concentrations observed during respiratory infections, activates Cl- secretion and inhibits Na+ absorption across the tracheal epithelium. It does so by binding to a neuraminidase-insensitive glycolipid, possibly asialo-GM1, triggering the release of ATP, which then acts in an autocrine fashion on apical P2Y receptors to produce the observed changes in ion transport. These findings indicate that fluid accumulation in the respiratory tract associated with parainfluenza virus infection is attributable, at least in part, to direct effects of the virus on ion transport by the respiratory epithelium. PMID- 15364904 TI - Organization and evolution of a gene-rich region of the mouse genome: a 12.7-Mb region deleted in the Del(13)Svea36H mouse. AB - Del(13)Svea36H (Del36H) is a deletion of approximately 20% of mouse chromosome 13 showing conserved synteny with human chromosome 6p22.1-6p22.3/6p25. The human region is lost in some deletion syndromes and is the site of several disease loci. Heterozygous Del36H mice show numerous phenotypes and may model aspects of human genetic disease. We describe 12.7 Mb of finished, annotated sequence from Del36H. Del36H has a higher gene density than the draft mouse genome, reflecting high local densities of three gene families (vomeronasal receptors, serpins, and prolactins) which are greatly expanded relative to human. Transposable elements are concentrated near these gene families. We therefore suggest that their neighborhoods are gene factories, regions of frequent recombination in which gene duplication is more frequent. The gene families show different proportions of pseudogenes, likely reflecting different strengths of purifying selection and/or gene conversion. They are also associated with relatively low simple sequence concentrations, which vary across the region with a periodicity of approximately 5 Mb. Del36H contains numerous evolutionarily conserved regions (ECRs). Many lie in noncoding regions, are detectable in species as distant as Ciona intestinalis, and therefore are candidate regulatory sequences. This analysis will facilitate functional genomic analysis of Del36H and provides insights into mouse genome evolution. PMID- 15364906 TI - Solution structure of a ubiquitin-like domain from tubulin-binding cofactor B. AB - Proper folding and assembly of tubulin alphabeta-heterodimers involves a stepwise progression mediated by a group of protein cofactors A through E. Upon release of the tubulin monomers from the chaperonin CCT, they are acted upon by each cofactor in the folding pathway through a unique combination of protein interaction domains. Three-dimensional structures have previously been reported for cofactor A and the C-terminal CAP-Gly domain of cofactor B (CoB). Here we report the NMR structure of the N-terminal domain of Caenorhabditis elegans CoB and show that it closely resembles ubiquitin as was recently postulated on the basis of bioinformatic analysis (Grynberg, M., Jaroszewski, L., and Godzik, A. (2003) BMC Bioinformatics 4, 46). CoB binds partially folded alpha-tubulin monomers, and a putative tubulin-binding motif within the N-terminal domain is identified from sequence and structure comparisons. Based on modeling of the homologous cofactor E ubiquitin-like domain, we hypothesize that cofactors B and E may associate via their beta-grasp domains in a manner analogous to the PB1 and caspase-activated deoxyribonuclease superfamily of protein interaction domains. PMID- 15364907 TI - Crystal structure of human kynurenine aminotransferase I. AB - The kynurenine pathway has long been regarded as a valuable target for the treatment of several neurological disorders accompanied by unbalanced levels of metabolites along the catabolic cascade, kynurenic acid among them. The irreversible transamination of kynurenine is the sole source of kynurenic acid, and it is catalyzed by different isoforms of the 5'-pyridoxal phosphate-dependent kynurenine aminotransferase (KAT). The KAT-I isozyme has also been reported to possess beta-lyase activity toward several sulfur- and selenium-conjugated molecules, leading to the proposal of a role of the enzyme in carcinogenesis associated with environmental pollutants. We solved the structure of human KAT-I in its 5'-pyridoxal phosphate and pyridoxamine phosphate forms and in complex with the competing substrate l-Phe. The enzyme active site revealed a striking crown of aromatic residues decorating the ligand binding pocket, which we propose as a major molecular determinant for substrate recognition. Ligand-induced conformational changes affecting Tyr(101) and the Trp(18)-bearing alpha-helix H1 appear to play a central role in catalysis. Our data reveal a key structural role of Glu(27), providing a molecular basis for the reported loss of enzymatic activity displayed by the equivalent Glu --> Gly mutation in KAT-I of spontaneously hypertensive rats. PMID- 15364908 TI - Cytoplasmic sequestration of HDAC7 from mitochondrial and nuclear compartments upon initiation of apoptosis. AB - Control of global histone acetylation status is largely governed by the opposing enzymatic activities of histone acetyltransferases and deacetylases (HDACs). HDACs were originally identified as modulators of nuclear histone acetylation status and have been linked to chromosomal condensation and subsequent gene repression. Accumulating evidence highlights HDAC modification of non-histone targets. Mitochondria were first characterized as intracellular organelles responsible for energy production through the coupling of oxidative phosphorylation to respiration. More recently, mitochondria have been implicated in programmed cell death whereby release of pro-apoptotic inner membrane space factors facilitates apoptotic progression. Here we describe the novel discovery that the nuclear encoded Class II human histone deacetylase HDAC7 localizes to the mitochondrial inner membrane space of prostate epithelial cells and exhibits cytoplasmic relocalization in response to initiation of the apoptotic cascade. These results highlight a previously unrecognized link between HDACs, mitochondria, and programmed cell death. PMID- 15364909 TI - The coxsackievirus and adenovirus receptor interacts with the multi-PDZ domain protein-1 (MUPP-1) within the tight junction. AB - The coxsackievirus and adenovirus receptor (CAR) is a component of the epithelial cell tight junction. In a yeast two-hybrid screen we identified the multi-PDZ domain protein MUPP1 as an interaction partner for the CAR cytoplasmic domain. CAR and MUPP1 were found to colocalize at the tight junction, to coprecipitate from epithelial cells, and to interact in vitro. The interaction was found to specifically involve the PDZ-binding motif within the CAR C terminus and MUPP1 PDZ domain 13. In transfected cells, CAR recruited MUPP1 to cell-cell contacts. The inhibition of CAR expression with small interfering RNA inhibited MUPP1 localization to the tight junction. The results indicated that CAR interacts with MUPP1 and is involved in MUPP1 recruitment to the tight junction. PMID- 15364910 TI - Phylogenetically conserved binding of specific K homology domain proteins to the 3'-untranslated region of the vertebrate middle neurofilament mRNA. AB - As axons mature, neurofilament-M (NF-M) expression rises, contributing to maturation of the axonal cytoskeleton and an expansion in axon caliber. This increase is partly due to a rise in NF-M mRNA stability. Such post transcriptional regulation is often mediated through the binding of specific proteins to the 3'-untranslated region (3'-UTR) of mRNAs. Vertebrate NF-M 3'-UTRs are remarkably well conserved, prompting us to test whether similar proteins bind the 3'-UTRs of different vertebrate NF-Ms. Identification of such proteins could lead to insights into the regulation of NF-M expression during development and in response to trauma or disease. Ultraviolet cross-linking analysis of proteins isolated from adult frog (Xenopus laevis), mouse, and rat brains revealed three ribonucleoprotein complexes (97, 70, and 47 kDa) that were present in all species and bound specifically to NF-M 3'-UTRs. Affinity purification of NF-M 3'-UTR binding proteins from rat brain followed by mass spectrometry and immunoprecipitation assays identified heterogeneous nuclear ribonucleoprotein (hnRNP) K and hnRNP E1 as the proteins forming the 70- and 47-kDa complexes, respectively. These RNA-binding proteins of the KH domain family recognize CU rich motifs identical to ones present in NF-M 3'-UTRs. Ultraviolet cross-linking assays performed on Xenopus embryos at different stages of neural development demonstrated that whereas hnRNP K binding occurred at all stages, hnRNP E binding occurred only at the most mature stages of axon development. Since hnRNP E is known to stabilize mRNAs, these results raise the hypothesis that these proteins may contribute to the increases in cytoplasmic levels of NF-M mRNA that accompany axonal maturation. PMID- 15364911 TI - Functional consequences of alpha-synuclein tyrosine nitration: diminished binding to lipid vesicles and increased fibril formation. AB - Previous studies have shown the presence of nitrated alpha-synuclein (alpha-syn) in human Lewy bodies and other alpha-syn inclusions. Herein, the effects of tyrosine nitration on alpha-syn fibril formation, lipid binding, chaperone-like function, and proteolytic degradation were systematically examined by employing chromatographically isolated nitrated monomeric, dimeric, and oligomeric alpha syn. Nitrated alpha-syn monomers and dimers but not oligomers accelerated the rate of fibril formation of unmodified alpha-syn when present at low concentrations. Immunoelectron microscopy revealed that nitrated monomers and dimers are incorporated into the fibrils. However, the purified nitrated alpha syn monomer by itself was unable to form fibrils. Nitration of the tyrosine residue at position 39 was largely responsible for decreased binding of nitrated monomeric alpha-syn to synthetic vesicles, which correlated with an impairment of the nitrated protein to adopt alpha-helical conformation in the presence of liposomes. The chaperone-like activity of alpha-syn was not inhibited by nitration or oxidation. Furthermore, the 20 S proteasome and calpain I degraded nitrated monomeric alpha-syn, although at a slower rate compared with control alpha-syn. Collectively, these data suggest that post-translational modification of alpha-syn by nitration can promote the formation of intracytoplasmic inclusions that constitute the hallmark of Parkinson disease and other synucleinopathies. PMID- 15364912 TI - Cytochrome bc(1) regulates the mitochondrial permeability transition by two distinct pathways. AB - The mitochondrial permeability transition (MPT) pore is a calcium-sensitive channel in the mitochondrial inner membrane that plays a crucial role in cell death. Here we show that cytochrome bc(1) regulates the MPT in isolated rat liver mitochondria and in CEM and HL60 cells by two independent pathways. Glutathione depletion activated the MPT via increased production of reactive oxygen species (ROS) generated by cytochrome bc(1). The ROS producing mechanism in cytochrome bc(1) involves movement of the "Rieske" iron-sulfur protein subunit of the enzyme complex, because inhibition of cytochrome bc(1) by pharmacologically blocking iron-sulfur protein movement completely abolished ROS production, MPT activation, and cell death. The classical inhibitor of the MPT, cyclosporine A, had no protective effect against MPT activation. In contrast, the calcium-activated, cyclosporine A-regulated MPT in rat liver mitochondria was also blocked with inhibitors of cytochrome bc(1). These results indicate that electron flux through cytochrome bc(1) regulates two distinct pathways to the MPT, one unregulated and involving mitochondrial ROS and the other regulated and activated by calcium. PMID- 15364913 TI - Functional organization of the sortilin Vps10p domain. AB - A Vps10p domain makes up the entire luminal part of Sortilin, and this type of domain is the hallmark of a new family of neuronal receptors that target a variety of ligands, including neurotrophins and neuropeptides. We have shown that two structural features of the Vps10p domain, the N-terminal propeptide and the C terminal segment of ten conserved cysteines (10CC), are key elements in the function of Sortilin. The propeptide has two functions. (i) It binds the mature part of Sortilin and prevents ligands in the biosynthetic pathway from binding to the uncleaved proreceptor, and (ii) it facilitates receptor transport in early Golgi compartments by a mechanism that does not depend on its ability to prevent ligand binding. In contrast, other Vps10p domain receptors, such as SorLA and SorCS3, do not need their propeptide for normal and swift processing. The 10CC segment constitutes an exchangeable module containing five conserved disulfide bridges, and using module-shuffling and truncations, we have shown that the 10CC segment is a major ligand-binding region in Sortilin. PMID- 15364914 TI - Multidrug resistance protein 4 (ABCC4)-mediated ATP hydrolysis: effect of transport substrates and characterization of the post-hydrolysis transition state. AB - Multidrug resistance protein 4 (MRP4/ABCC4), transports cyclic nucleoside monophosphates, nucleoside analog drugs, chemotherapeutic agents, and prostaglandins. In this study we characterize ATP hydrolysis by human MRP4 expressed in insect cells. MRP4 hydrolyzes ATP (Km, 0.62 mm), which is inhibited by orthovanadate and beryllium fluoride. However, unlike ATPase activity of P glycoprotein, which is equally sensitive to both inhibitors, MRP4-ATPase is more sensitive to beryllium fluoride than to orthovanadate. 8-Azido[alpha-32P]ATP binds to MRP4 (concentration for half-maximal binding approximately 3 microm) and is displaced by ATP or by its non-hydrolyzable analog AMPPNP (concentrations for half-maximal inhibition of 13.3 and 308 microm). MRP4 substrates, the prostaglandins E1 and E2, stimulate ATP hydrolysis 2- to 3-fold but do not affect the Km for ATP. Several other substrates, azidothymidine, 9-(2 phosphonylmethoxyethyl)adenine, and methotrexate do not stimulate ATP hydrolysis but inhibit prostaglandin E2-stimulated ATP hydrolysis. Although both post hydrolysis transition states MRP4.8-azido[alpha-32P]ADP.Vi and MRP4.8-azido[alpha 32P]ADP.beryllium fluoride can be generated, nucleotide trapping is approximately 4-fold higher with beryllium fluoride. The divalent cations Mg2+ and Mn2+ support comparable levels of nucleotide binding, hydrolysis, and trapping. However, Co2+ increases 8-azido[alpha-32P]ATP binding and beryllium fluoride-induced 8 azido[alpha-32P]ADP trapping but does not support steady-state ATP hydrolysis. ADP inhibits basal and prostaglandin E2-stimulated ATP hydrolysis (concentrations for half-maximal inhibition 0.19 and 0.25 mm, respectively) and beryllium fluoride-induced 8-azido[alpha-32P]ADP trapping, whereas Pi has no effect up to 20 mm. In aggregate, our results demonstrate that MRP4 exhibits substrate stimulated ATP hydrolysis, and we propose a kinetic scheme suggesting that ADP release from the post-hydrolysis transition state may be the rate-limiting step during the catalytic cycle. PMID- 15364915 TI - Protein kinase C betaII regulates Akt phosphorylation on Ser-473 in a cell type- and stimulus-specific fashion. AB - Akt (= protein kinase B), a subfamily of the AGC serine/threonine kinases, plays critical roles in survival, proliferation, glucose metabolism, and other cellular functions. Akt activation requires the recruitment of the enzyme to the plasma membrane by interacting with membrane-bound lipid products of phosphatidylinositol 3-kinase. Membrane-bound Akt is then phosphorylated at two sites for its full activation; Thr-308 in the activation loop of the kinase domain is phosphorylated by 3-phosphoinositide-dependent kinase-1 (PDK1) and Ser 473 in the C-terminal hydrophobic motif by a putative kinase PDK2. The identity of PDK2 has been elusive. Here we present evidence that conventional isoforms of protein kinase C (PKC), particularly PKCbetaII, can regulate Akt activity by directly phosphorylating Ser-473 in vitro and in IgE/antigen-stimulated mast cells. By contrast, PKCbeta is not required for Ser-473 phosphorylation in mast cells stimulated with stem cell factor or interleukin-3, in serum-stimulated fibroblasts, or in antigen receptor-stimulated T or B lymphocytes. Therefore, PKCbetaII appears to work as a cell type- and stimulus-specific PDK2. PMID- 15364916 TI - Melanocortin-independent effects of leptin on hepatic glucose fluxes. AB - Leptin and insulin share some hypothalamic signaling molecules, but their central administration induces different effects on hepatic glucose fluxes. Acute insulin infusion in the third cerebral ventricle inhibits endogenous glucose production (GP), whereas acute leptin infusion stimulates gluconeogenesis but does not alter GP because of a compensatory decrease in glycogenolysis. Because melanocortin agonists also stimulate hepatic gluconeogenesis, here we examined whether central melanocortin blockade modifies the acute effects of leptin on GP, on gluconeogenesis, on glycogenolysis, and/or on the hepatic expression of the gluconeogenic enzymes glucose-6-phosphatase (Glc-6-Pase) and phosphoenolpyruvate carboxykinase (PEPCK). Systemic or central administration of leptin alone did not alter GP, despite increasing both the rate of gluconeogenesis and the expression of Glc-6-Pase and PEPCK. When activation of the central melanocortin pathway was prevented, the effects of leptin on gluconeogenesis, Glc-6-Pase, and PEPCK were abolished, and a marked suppression of glycogenolysis resulted in decreased GP. We conclude that leptin regulates hepatic glucose fluxes through a melanocortin dependent pathway leading to stimulation of gluconeogenesis and a melanocortin independent pathway causing inhibition of GP and glycogenolysis. PMID- 15364917 TI - 4-cyanopyridine, a versatile spectroscopic probe for cytochrome P450 BM3. AB - The nitrogenous pi -acceptor ligand 4-cyanopyridine (4CNPy) exhibits reversible ligation to ferrous heme in the flavocytochrome P450 BM3 (Kd=1.8 microm for wild type P450 BM3) via its pyridine ring nitrogen. The reduced P450-4CNPy adduct displays unusual spectral properties that provide a useful spectroscopic handle to probe particular aspects of this P450. 4CNPy is competitively displaced upon substrate binding, allowing a convenient route to the determination of substrate dissociation constants for ferrous P450 highlighting an increase in P450 substrate affinity on heme reduction. For wild type P450 BM3, Kd(red)(laurate)=82.4 microm (cf. Kd(ox)=364 microm). In addition, an unusual spectral feature in the red region of the absorption spectrum of the reduced P450 4CNPy adduct is observed that can be assigned as a metal-to-ligand charge transfer (MLCT). It was discovered that the energy of this MLCT varies linearly with respect to the P450 heme reduction potential. By studying the energy of this MLCT for a series of BM3 active site mutants with differing reduction potential (Em), the relationship EMLCT + (3.53 x = Em 17,005 cm)(-1) was derived. The use of this ligand thus provides a quick and accurate method for predicting the heme reduction potentials of a series of P450 BM3 mutations using visible spectroscopy, without the requirement for redox potentiometry. PMID- 15364918 TI - Lateral diffusion of inositol 1,4,5-trisphosphate receptor type 1 is regulated by actin filaments and 4.1N in neuronal dendrites. AB - Inositol 1,4,5-trisphosphate receptor type1 (IP3R1) plays an important role in neuronal functions; however, the lateral diffusion of IP3R1 on the endoplasmic reticulum membrane and its regulation in the living neurons remain unknown. We expressed green fluorescent protein-tagged IP3R1 in cultured rat hippocampal neurons and observed the lateral diffusion by the fluorescence recovery after photobleaching technique. IP3R1 showed lateral diffusion with an effective diffusion constant of approximately 0.3 microm2/s. Depletion of actin filaments increased the diffusion constant of IP3R1, suggesting that the diffusion of IP3R1 is regulated negatively through actin filaments. We also found that protein 4.1N, which binds to IP3R1 and contains an actin-spectrin-binding region, was responsible for this actin regulation of the IP3R1 diffusion constant. Overexpression of dominant-negative 4.1N and blockade of 4.1N binding to IP3R1 increased the IP3R1 diffusion constant. The diffusion of IP3R type 3 (IP3R3), one of the isoforms of IP3Rs lacking the binding ability to 4.1N, was not dependent on actin filaments but became dependent on actin filaments after the addition of a 4.1N-binding sequence. These data suggest that 4.1N serves as a linker protein between IP3R1 and actin filaments. This actin filament-dependent regulation of IP3R1 diffusion may be important for the spatiotemporal regulation of intracellular Ca2+ signaling. PMID- 15364919 TI - Protein kinase C Theta inhibits insulin signaling by phosphorylating IRS1 at Ser(1101). AB - Obesity and stress inhibit insulin action by activating protein kinases that enhance serine phosphorylation of IRS1 and have been thus associated to insulin resistance and the development of type II diabetes. The protein kinase C (PKC) is activated by free-fatty acids, and its activity is higher in muscle from obese diabetic patients. However, a molecular link between PKC and insulin resistance has not been defined yet. Here we show that PKC phosphorylates IRS1 at serine 1101 blocking IRS1 tyrosine phosphorylation and downstream activation of the Akt pathway. Mutation of Ser(1101) to alanine makes IRS1 insensitive to the effect of PKC and restores insulin signaling in culture cells. These results provide a novel mechanism linking the activation of PKC to the inhibition of insulin signaling. PMID- 15364920 TI - CD22 is a functional ligand for SH2 domain-containing protein-tyrosine phosphatase-1 in primary T cells. AB - The intracellular Src homology 2 (SH2) domain-containing protein-tyrosine phosphatase (SHP-1) has been characterized as a negative regulator of T cell function, contributing to the definition of T cell receptor signaling thresholds in developing and peripheral mouse T lymphocytes. The activation of SHP-1 is achieved through the engagement of its tandem SH2 domains by tyrosine phosphorylated proteins; however, the identity of the activating ligand(s) for SHP-1, within mouse primary T cells, is presently unresolved. The identification of SHP-1 ligand(s) in primary T cells would provide crucial insight into the molecular mechanisms by which SHP-1 contributes to in vivo thresholds for T cell activation. Here we present a combination of biochemical and yeast genetic analyses indicating CD22 to be a T cell ligand for the SHP-1 SH2 domains. Based on these observations we have confirmed that CD22 is indeed expressed on mouse primary T cells and capable of associating with SHP-1. Significantly, CD22 deficient T cells demonstrate enhanced proliferation in response to anti-CD3 or allogeneic stimulation. Furthermore, the co-engagement of CD3 and CD22 results in a raising of TCR signaling thresholds hence demonstrating a previously unsuspected functional role for CD22 in primary T cells. PMID- 15364921 TI - Aromatic amino acid transporter AAT-9 of Caenorhabditis elegans localizes to neurons and muscle cells. AB - The Caenorhabditis elegans genome encodes nine homologues of mammalian glycoprotein-associated amino acid transporters. Two of these C. elegans proteins (AAT-1 and AAT-3) have been shown to function as catalytic subunits (light chains) of heteromeric amino acid transporters. These proteins need to associate with a glycoprotein heavy chain subunit (ATG-2) to reach the cell surface in a manner similar to that of their mammalian homologues. AAT-1 and AAT-3 contain a cysteine residue in the second putative extracellular loop through which a disulfide bridge can form with a heavy chain. In contrast, six C. elegans members of this family (AAT-4 to AAT-9) lack such a cysteine residue. We show here that one of these transporter proteins, AAT-9, reaches the cell surface in Xenopus oocytes without an exogenous heavy chain and that it functions as an exchanger of aromatic amino acids. Two-electrode voltage clamp experiments demonstrate that AAT-9 displays a substrate-activated conductance. Immunofluorescence shows that it is expressed close to the pharyngeal bulbs within C. elegans neurons. The selective expression of an aat-9 promoter-green fluorescent protein construct in several neurons of this region and in wall muscle cells around the mouth supports and extends these localization data. Taken together, the results show that AAT-9 is expressed in excitable cells of the nematode head and pharynx in which it may provide a pathway for aromatic amino acid transport. PMID- 15364922 TI - Inhibition of geranylgeranylation mediates the effects of 3-hydroxy-3 methylglutaryl (HMG)-CoA reductase inhibitors on microglia. AB - Inflammatory responses involving microglia, the resident macrophages of the brain, are thought to contribute importantly to the progression of Alzheimer's disease (AD) and possibly other neurodegenerative disorders. The present study tested whether the mevalonate-isoprenoid biosynthesis pathway, which affects inflammation in many types of tissues, tonically regulates microglial activation. This question takes on added significance given the potential use of statins, drugs that block the rate-limiting step (3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase)) in mevalonate and cholesterol synthesis, in AD treatment. Both mevastatin and simvastatin caused a concentration- and time dependent activation of microglia in cultured rat hippocampal slices. This response consisted of a transformation of the cells from a typical resting configuration to an amoeboid, macrophage-like morphology, increased expression of a macrophage antigen, and up-regulation of the cytokine tumor necrosis factor alpha. Evidence for proliferation was also obtained. Statin-induced microglial changes were blocked by mevalonate but not by cholesterol, indicating that they were probably due to suppression of isoprenoid synthesis. In accord with this, the statin effects were absent in slices co-incubated with geranylgeranyl pyrophosphate, a mevalonate product that provides for the prenylation of Rho GTPases. Finally, PD98089, a compound that blocks activation of extracellularly regulated kinases1/2, suppressed statin-induced up-regulation of tumor necrosis factor-alpha but had little effect on microglial transformation. These results suggest that 1) the mevalonate-isoprenoid pathway is involved in regulating microglial morphology and in controlling expression of certain cytokines and 2) statins have the potential for enhancing a component of AD with uncertain relationships to other features of the disease. PMID- 15364923 TI - Optimal lysophosphatidic acid-induced DNA synthesis and cell migration but not survival require intact autophosphorylation sites of the epidermal growth factor receptor. AB - Lysophosphatidic acid (LPA)-elicited transphosphorylation of receptor tyrosine kinases has been implicated in mediating extracellular signal-regulated kinase (ERK) 1/2 activation, which is necessary for LPA-induced cell proliferation, migration, and survival. B82L cells lack epidermal growth factor receptor (EGFR) but express LPA(1-3), platelet-derived growth factor (PDGF), ErbB2, and insulin like growth factor receptor transcripts, yet LPA caused no detectable transphosphorylation of these receptor tyrosine kinases. LPA equally protected B82L cells, or transfectants expressing EGFR, the kinase dead EGFR(K721A), EGFR(Y5F) receptor mutant, which lacks five autophosphorylation sites, or EGFR(Y845F), which lacks the Src phosphorylation site from tumor necrosis factor alpha-induced apoptosis. In contrast, LPA-elicited DNA synthesis and migration were augmented in cells expressing EGFR, EGFR(K721A), or EGFR(Y845F), but not EGFR(Y5F), although the PDGF responses were indistinguishable. LPA-induced transphosphorylation of the EGFR, ErbB2, or PDGF receptor was not required for its antiapoptotic effect. EGFR with or without intrinsic kinase activity or without the Src-phosphorylation site augmented, but was not required for, LPA elicited cell proliferation or migration. In B82L cells, augmentation of these two LPA responses required intact autophosphorylation sites because among the four EGFR mutants, only cells expressing the EGFR(Y5F) mutant showed no enhancement. In EGFR(Y5F)-expressing cells, LPA failed to elicit tyrosine phosphorylation of Src homologous and collagen protein (SHC) and caused only a modest increase in ERK1/2 phosphorylation similar to that in wild-type B82L cells. The present data pinpoint the lack of importance of the intrinsic kinase activity in contrast to the importance of autophosphorylation sites of the EGFR for SHC phosphorylation in the enhancement of select ERK1/2-dependent LPA responses. PMID- 15364924 TI - Pseudophosphorylation and glycation of tau protein enhance but do not trigger fibrillization in vitro. AB - Alzheimer's disease is defined in part by the intraneuronal aggregation of tau protein into filamentous lesions. The pathway is accompanied by posttranslational modifications including phosphorylation and glycation, each of which has been shown to promote tau fibrillization in vitro when present at high stoichiometry. To clarify the site-specific impact of posttranslational modification on tau fibrillization, the ability of recombinant full-length four repeat tau protein (htau40) and 11 pseudophosphorylation mutants to fibrillize in the presence of anionic inducer was assayed in vitro using transmission electron microscopy and laser light scattering assays. Tau glycated with d-glucose was examined as well. Both glycated tau and pseudophosphorylation mutants S199E, T212E, S214E, double mutant T212E/S214E, and triple mutant S199E/S202E/T205E yielded increased filament mass at equilibrium relative to wild-type tau. Increases in filament mass correlated strongly with decreases in critical concentration, indicating that both pseudophosphorylation and glycation promoted fibrillization by shifting equilibrium toward the fibrillized state. Analysis of reaction time courses further revealed that increases in filament mass were not associated with reduced lag times, indicating that these posttranslational modifications did not promote filament nucleation. The results suggest that site-specific posttranslational modifications can stabilize filaments once they nucleate, and thereby support their accumulation at low intracellular tau concentrations. PMID- 15364925 TI - Antigen stability controls antigen presentation. AB - We investigated whether protein stability controls antigen presentation using a four disulfide-containing snake toxin and three derivatives carrying one or two mutations (L1A, L1A/H4Y, and H4Y). These mutations were anticipated to increase (H4Y) or decrease (L1A) the antigen non-covalent stabilizing interactions, H4Y being naturally and frequently observed in neurotoxins. The chemically synthesized derivatives shared similar three-dimensional structure, biological activity, and T epitope pattern. However, they displayed differential thermal unfolding capacities, ranging from 65 to 98 degrees C. Using these differentially stable derivatives, we demonstrated that antigen stability controls antigen proteolysis, antigen processing in antigen-presenting cells, T cell stimulation, and kinetics of expression of T cell determinants. Therefore, non-covalent interactions that control the unfolding capacity of an antigen are key parameters in the efficacy of antigen presentation. By affecting the stabilizing interaction network of proteins, some natural mutations may modulate the subsequent T-cell stimulation and might help microorganisms to escape the immune response. PMID- 15364926 TI - Interactions between extracellular signal-regulated protein kinase 1, 14-3 3epsilon, and heat shock factor 1 during stress. AB - Cytoprotection during the heat shock response is a complex phenomenon involving multiple inducible mechanisms. We have examined the interaction of two key molecular components in the response, heat shock transcription factor 1 (HSF1) and extracellular signal regulated protein kinase (ERK). Whereas both HSF1 and ERK are required to protect cells against apoptosis, ERK activation is paradoxically antagonistic to trans-activation of hsp promoters by HSF1 and HSP accumulation during heat shock. We have found that the two pathways interact directly and that heat shock causes the physical association of ERK1 with HSF1, an interaction that promotes the kinase activity of ERK in heat-shocked cells. ERK activation results in the recruitment of the phosphoserine binding protein 14 3-3epsilon in a manner dependent on previous HSF1 phosphorylation by ERK. The effects of 14-3-3epsilon binding on HSF1 were complex, however, depending on extracellular conditions, in that HSF1-14-3-3 binding at 37 degrees C led to the cytoplasmic sequestration and repression of HSF1, whereas heat shock overrode these effects and caused quantitative nuclear localization of HSF1. Although the effects of 14-3-3epsilon binding to HSF1 were overridden acutely by stress, during recovery from heat shock, 14-3-3epsilon association again led to enhanced cytoplasmic localization of HSF1, implicating a role for ERK/14-3-3epsilon in HSF1 deactivation in recovering cells. Association of HSF1 with ERK and 14-3 3epsilon during heat shock may thus modulate the amplitude of the response and lead to efficient termination of HSP expression on resumption of growth conditions. PMID- 15364927 TI - Negative regulation of p53 functions by Daxx and the involvement of MDM2. AB - In normal cells p53 activity is tightly controlled and MDM2 is a known negative regulator. Here we show that via its acidic domain, Daxx binds to the COOH terminal domain of p53, whose positive charges are critical for this interaction, as Lys to Arg mutations preserved, but Lys to Ala or Ser to Glu mutations abolished Daxx-p53 interaction. These results thus implicate acetylation and phosphorylation of p53 in regulating its binding to Daxx. Interestingly, whereas Daxx did not bind to p53 in cells as assessed by immunoprecipitation, MDM2 expression restored p53-Daxx interaction, and this correlated with deacetylation of p53. In p53/MDM2-null mouse embryonic fibroblasts (DKO MEF), Daxx repressed p53 target promoters whose p53-binding elements were required for the repression. Coexpression of Daxx and MDM2 led to further repression. p53 expression in DKO MEF induced apoptosis and Daxx expression relieved this effect. Similarly, in HCT116 cells, Daxx conferred striking resistance to 5-fluorouracil-induced apoptosis. As p53 is required for 5-fluorouracil-induced cell death, our data show that Daxx can suppress cell death induced by p53 overexpression and p53 dependent stress response. Collectively, our data reveal Daxx as a novel negative regulator of p53. Importantly, posttranslational modifications of p53 inhibit Daxx-p53 interaction, thereby relieving negative regulation of p53 by Daxx. PMID- 15364928 TI - Rafts can trigger contact-mediated secretion of bacterial effectors via a lipid based mechanism. AB - Infection by the Gram-negative bacterial pathogen Shigella flexneri depends on its ability to invade host cells. Bacterial engulfment requires a functional type III secretion system (TTSS) allowing the translocation into host cells of bacterial effectors that activate cell-signaling cascades. We demonstrated previously that specialized lipid membrane domains enriched in cholesterol and sphingolipids (rafts) are involved during early steps of invasion, namely in binding and host cell entry. In this study, we addressed the issue of contact mediated secretion by the TTSS. We show that contact-mediated and TTSS-induced hemolysis depend on the presence of cholesterol on the host cell surface. We found that purified detergent resistant membranes were able to activate TTSS. Finally, we found that artificial liposomes, devoid of proteins, were able to activate the TTSS but only when their composition mimicked that of lipid rafts. Altogether, these data indicate that specific lipid packing can trigger contact mediated secretion by S. flexneri. PMID- 15364929 TI - Identification, cloning, expression, and purification of three novel human calcium-independent phospholipase A2 family members possessing triacylglycerol lipase and acylglycerol transacylase activities. AB - Genetic knockout of hormone-sensitive lipase in mice has implicated the presence of other intracellular triacylglycerol (TAG) lipases mediating TAG hydrolysis in adipocytes. Despite intense interest in these TAG lipases, their molecular identities thus far are largely unknown. Sequence data base searches for proteins containing calcium-independent phospholipase A2 (iPLA2) dual signature nucleotide ((G/A)XGXXG) and lipase (GXSXG) consensus sequence motifs identified a novel subfamily of three putative iPLA2/lipase family members designated iPLA2epsilon, iPLA2zeta, and iPLA2eta (previously named adiponutrin, TTS-2.2, and GS2, respectively) of previously unknown catalytic function. Herein we describe the cloning, heterologous expression, and affinity purification of the three human isoforms of this iPLA2 subfamily in Sf9 cells, and we demonstrate that each possesses abundant TAG lipase activity. Moreover, iPLA2epsilon, iPLA2zeta, and iPLA2eta also possess acylglycerol transacylase activity utilizing mono-olein as an acyl donor which, in the presence of mono-olein or diolein acceptors, results in the synthesis of diolein and triolein, respectively. (E)-6-(Bromomethylene)-3 (1-naphthalenyl)-2H-tetrahydropyran-2-one, a mechanism-based suicide substrate inhibitor of all known iPLA2s, inhibits the triglyceride lipase activity of each of the three isoforms similarly (IC50=0.1-0.5 microm). Quantitative PCR revealed dramatically increased expression of iPLA2epsilon and iPLA2zeta transcripts during the hormone-induced differentiation of 3T3-L1 cells into adipocytes and identified the presence of all three iPLA2 isoforms in human SW872 liposarcoma cells. Collectively, these results identify three novel TAG lipases/acylglycerol transacylases that likely participate in TAG hydrolysis and the acyl-CoA independent transacylation of acylglycerols, thereby facilitating energy mobilization and storage in adipocytes. PMID- 15364930 TI - Alternative splicing of Staufen2 creates the nuclear export signal for CRM1 (Exportin 1). AB - Mammalian Staufen2 (Stau2), a brain-specific double-stranded RNA-binding protein, is involved in the localization of mRNA in neurons. To gain insights into the function of Stau2, the subcellular localization of Stau2 isoforms fused to the green fluorescence protein was examined. Fluorescence microscopic analysis showed that Stau2 functions as a nucleocytoplasmic shuttle protein. The nuclear export of the 62-kDa isoform of Stau2 (Stau2(62)) is mediated by the double-stranded RNA binding domain 3 (RBD3) because a mutation to RBD3 led to nuclear accumulation. On the other hand, the shorter isoform of Stau2, Stau2(59), is exported from the nucleus by two distinct pathways, one of which is RBD3-mediated and the other of which is CRM1 (exportin 1)-dependent. The nuclear export signal recognized by CRM1 was found to be located in the N-terminal region of Stau2(59). These results suggest that Stau2 may carry a variety of RNAs out of the nucleus, using the two export pathways. The present study addresses the issue of why plural Stau2 isoforms are expressed in neurons. PMID- 15364931 TI - A C-terminal Myb extension domain defines a novel family of double-strand telomeric DNA-binding proteins in Arabidopsis. AB - Little is known about the protein composition of plant telomeres. We queried the Arabidopsis thaliana genome data base in search of genes with similarity to the human telomere proteins hTRF1 and hTRF2. hTRF1/hTRF2 are distinguished by the presence of a single Myb-like domain in their C terminus that is required for telomeric DNA binding in vitro. Twelve Arabidopsis genes fitting this criterion, dubbed TRF-like (TRFL), fell into two distinct gene families. Notably, TRFL family 1 possessed a highly conserved region C-terminal to the Myb domain called Myb-extension (Myb-ext) that is absent in TRFL family 2 and hTRF1/hTRF2. Immunoprecipitation experiments revealed that recombinant proteins from TRFL family 1, but not those from family 2, formed homodimers and heterodimers in vitro. DNA binding studies with isolated C-terminal fragments from TRFL family 1 proteins, but not family 2, showed specific binding to double-stranded plant telomeric DNA in vitro. Removal of the Myb-ext domain from TRFL1, a family 1 member, abolished DNA binding. However, when the Myb-ext domain was introduced into the corresponding region in TRFL3, a family 2 member, telomeric DNA binding was observed. Thus, Myb-ext is required for binding plant telomeric DNA and defines a novel class of proteins in Arabidopsis. PMID- 15364932 TI - Critical role for protein phosphatase 2A heterotrimers in mammalian cell survival. AB - The predominant forms of protein phosphatase 2A (PP2A), one of the major Ser/Thr phosphatases, are dimers of catalytic (C) and scaffolding (A) subunits and trimers with an additional variable regulatory subunit. In mammals, catalytic and scaffolding subunits are encoded by two genes each (alpha/beta), whereas three gene families (B, B', and B'') with a total of 12 genes contribute PP2A regulatory subunits. We generated stable PC12 cell lines in which the major scaffolding Aalpha subunit can be knocked down by inducible RNA interference (RNAi) to study its role in cell viability. Aalpha RNAi decreased total PP2A activity as well as protein levels of C, B, and B' but not B'' subunits. Inhibitor experiments indicate that monomeric C and B subunits are degraded by the proteosome. Knock-down of Aalpha triggered cell death by redundant apoptotic and non-apoptotic mechanisms because the inhibition of RNAi-associated caspase activation failed to stall cell death. PP2A holoenzymes positively regulate survival kinase signaling, because RNAi reduced basal and epidermal growth factor stimulated Akt phosphorylation. RNAi-resistant Aalpha cDNAs rescued RNAi-induced loss of the C subunit, and Aalpha point mutants prevented regulatory subunit degradation as predicted from each mutant's binding specificity. In transient, stable, and stable-inducible rescue experiments, both wild-type Abeta and Aalpha mutants capable of binding to at least one family of regulatory subunits were able to delay Aalpha RNAi-induced death of PC12 cells. However, only the expression of wild-type Aalpha restored viability completely. Thus, heterotrimeric PP2A holoenzymes containing the Aalpha subunit and members of all three regulatory subunit families are necessary for mammalian cell viability. PMID- 15364933 TI - The Nox family of NAD(P)H oxidases: host defense and beyond. PMID- 15364934 TI - Protein phosphatase 4 is a positive regulator of hematopoietic progenitor kinase 1. AB - Hematopoietic progenitor kinase 1 (HPK1) is a hematopoietic specific mammalian Ste20-like protein kinase and has been implicated in many cellular signaling pathways including T cell receptor (TCR) signaling. However, little is known about the in vivo regulation of HPK1. We present evidence that HPK1 is positively regulated by protein phosphatase 4 (PP4; also called PPX and PPP4), a serine/threonine phosphatase. We found that PP4 interacted with HPK1 and that the proline-rich region of HPK1 was necessary and sufficient for this interaction. We also found that PP4 had phosphatase activity toward HPK1 in vivo and that co transfection of PP4 with HPK1 resulted in specific kinase activation of HPK1. Moreover, we found that the PP4-induced HPK1 kinase activation was accompanied by an increase in protein expression of HPK1. Pulse-chase analysis showed that PP4 increased the half-life of HPK1. Further studies showed that HPK1 was subject to regulation by ubiquitination and ubiquitin-targeted degradation and that PP4 inhibited HPK1 ubiquitination. In addition, we found that TCR stimulation enhanced the PP4-HPK1 interaction and that wild-type PP4 enhanced, whereas a phosphatase-dead PP4 mutant inhibited, TCR-induced activation of HPK1 in Jurkat T cells. Combined with the observation that PP4 enhanced HPK1-induced JNK activation, our studies identify PP4 as a positive regulator for HPK1 and the HPK1-JNK signaling pathway. PMID- 15364935 TI - Regulation of cardiac IKs potassium current by membrane phosphatidylinositol 4,5 bisphosphate. AB - Regulation of the slowly activating component of delayed rectifier K+ current (IKs) by membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PtdIns (4,5)P2) was examined in guinea pig atrial myocytes using the whole-cell patch clamp method. IKs was elicited by depolarizing voltage steps given from a holding potential of -50 mV, and the effect of various test reagents on IKs was assessed by measuring the amplitude of tail current elicited upon return to the holding potential following a 2-s depolarization to +30 mV. Intracellular application of 50 microM wortmannin through a recording pipette evoked a progressive increase in IKs over a 10-15-min period to 208.5 +/- 14.6% (n = 9) of initial magnitude obtained shortly after rupture of the patch membrane. Intracellular application of anti-PtdIns(4,5)P2 monoclonal antibody also increased the amplitude of IKs to 198.4 +/- 19.9% (n = 5). In contrast, intracellular loading with exogenous PtdIns(4,5)P2 at 10 and 100 mum produced a marked decrease in the amplitude of IKs to 54.3 +/- 3.8% (n = 5) and 44.8 +/- 8.2% (n = 5), respectively. Intracellular application of neomycin (50 microM) or aluminum (50 microM) evoked an increase in the amplitude of IKs to 161.0 +/- 13.5% (n = 4) and 150.0 +/- 8.2% (n = 4), respectively. These results strongly suggest that IKs channel is inhibited by endogenous membrane PtdIns(4,5)P2 through the electrostatic interaction with the negatively charged head group on PtdIns(4,5)P2. Potentiation of IKs by P2Y receptor stimulation with 50 microM ATP was almost totally abolished when PtdIns(4,5)P2 was included in the pipette solution, suggesting that depletion of membrane PtdIns(4,5)P2 is involved in the potentiation of IKs by P2Y receptor stimulation. Thus, membrane PtdIns(4,5)P2 may act as an important physiological regulator of IKs in guinea pig atrial myocytes. PMID- 15364936 TI - Recognition of phosphodegron motifs in human cyclin E by the SCF(Fbw7) ubiquitin ligase. AB - Turnover of cyclin E is controlled by SCF(Fbw7). Three isoforms of Fbw7 are produced by alternative splicing. Whereas Fbw7alpha and -gamma are nuclear and the beta-isoform is cytoplasmic in 293T cells, all three isoforms induce cyclin E destruction in an in vivo degradation assay. Cyclin E is phosphorylated on Thr(62), Ser(88), Ser(372), Thr(380), and Ser(384) in vivo. To examine the roles of phosphorylation in cyclin E turnover, a series of alanine point mutations in each of these sites were analyzed for Fbw7-driven degradation. As expected, mutation of the previously characterized residue Thr(380) to alanine led to profound defects of cyclin E turnover, and largely abolished association with Fbw7. Mutation of Thr(62) to alanine led to a dramatic reduction in the extent of Thr(380) phosphorylation, suggesting an indirect effect of this mutation on cyclin E turnover. Nevertheless, phosphopeptides centered at Thr(62) associated with Fbw7, and residual binding of cyclin E(T380A) to Fbw7 was abolished upon mutation of Thr(62), suggesting a minor role for this residue in direct association with Fbw7. Mutation of Ser(384) to alanine also rendered cyclin E resistant to degradation by Fbw7, with the largest effects being observed with Fbw7beta. Cyclin E(S384A) associated more weakly with Fbw7alpha and -beta isoforms but was not defective in Thr(380) phosphorylation. Analysis of the localization of cyclin E mutant proteins indicated selective accumulation of cyclin E(S384A) in the nucleus, which may contribute to the inability of cytoplasmic Fbw7beta to promote turnover of this cyclin E mutant protein. PMID- 15364937 TI - Catalytic domain crystal structure of protein kinase C-theta (PKCtheta). AB - A member of the novel protein kinase C (PKC) subfamily, PKC, is an essential component of the T cell synapse and is required for optimal T cell activation and interleukin-2 production. Selective involvement of PKC in TCR signaling makes this enzyme an attractive therapeutic target in T cell-mediated disease processes. In this report we describe the crystal structure of the catalytic domain of PKC at 2.0-A resolution. Human recombinant PKC kinase domain was expressed in bacteria as catalytically active phosphorylated enzyme and co crystallized with its subnanomolar, ATP site inhibitor staurosporine. The structure follows the classic bilobal kinase fold and shows the enzyme in its active conformation and phosphorylated state. Inhibitory interactions between conserved features of staurosporine and the ATP-binding cleft are accompanied by closing of the glycine-rich loop, which also maintains an inhibitory arrangement by blocking the phosphate recognition subsite. The two major phosphorylation sites, Thr-538 in the activation loop and Ser-695 in the hydrophobic motif, are both occupied in the structure, playing key roles in stabilizing active conformation of the enzyme and indicative of PKC autocatalytic phosphorylation and activation during bacterial expression. The PKC-staurosporine complex represents the first kinase domain crystal structure of any PKC isotypes to be determined and as such should provide valuable insight into PKC specificity and into rational drug design strategies for PKC selective leads. PMID- 15364938 TI - alpha-thrombin rapidly induces tyrosine phosphorylation of a novel, 74-78-kDa stress response protein(s) in lung fibroblast cells. AB - We demonstrated previously that exposure of CCL39 lung fibroblasts to alpha thrombin rapidly inhibits interleukin 6-induced tyrosine phosphorylation of signal transducers and activators of transcription 3 (Stat3). While studying the cross-talk between alpha-thrombin and interleukin 6, we observed that the phospho specific (tyrosine) anti-Stat3 antibody specifically cross-reacted with a 74-78 kDa protein(s) in alpha-thrombin-treated cells. In this study, we demonstrate that in alpha-thrombin-treated CCL39 cells, the 74-78-kDa protein(s) rapidly undergoes tyrosine phosphorylation. The phosphorylation by alpha-thrombin was detected as early as 5 min and reached a maximum at 15 min; however, low levels were present at 2 h. alpha-Thrombin receptor agonist peptide (SFLLRN) induced its tyrosine phosphorylation, suggesting that alpha-thrombin mediates the effects via protease-activated receptor type 1. Anti-Stat3 antibodies specific to different regions of Stat3 failed to recognize the 74-78-kDa protein(s), suggesting that it is unrelated to Stat3. Cell fractionation experiments showed that it is localized to the cytoplasm. Mass spectrometric analysis of the immunoprecipitated protein showed that the 74-78-kDa protein(s) is related to glucose-regulated protein 75 (GRP-75), a member of the heat shock/stress-response protein family. Consistent with these data, we observed tyrosine phosphorylation of GRP-75 in alpha-thrombin treated cells. Exposure of cells to pervanadate, a stress-inducing agent, stimulated its tyrosine phosphorylation; however, cytokines and growth factors were ineffective. This is the first report of tyrosine phosphorylation of GRP-75 related stress protein(s) by alpha-thrombin and suggests that this pathway may contribute to the ability of alpha-thrombin to prevent apoptosis in cells exposed to stress or in the injured tissue. PMID- 15364939 TI - Differential modes of transfer RNASer recognition in Methanosarcina barkeri. AB - Two dissimilar seryl-transfer RNA (tRNA) synthetases (SerRSs) exist in Methanosarcina barkeri, one of bacterial type and the other resembling SerRSs present only in some methanogenic archaea. To investigate the requirements of these enzymes for tRNASer recognition, serylation of variant transcripts of M. barkeri tRNASer was kinetically analyzed in vitro with pure enzyme preparations. Characteristically for the serine system, the length of the variable arm was shown to be crucial for both enzymes, as was the identity of the discriminator base (G73). Moreover, a novel determinant for the specific tRNASer recognition was identified as the anticodon stem base pair G30:C40; its contribution to the efficiency of serylation was remarkable for both SerRSs. However, despite these similarities, the two SerRSs do not possess a uniform mode of tRNASer recognition, and additional determinants are necessary for serylation specificity by the methanogenic enzyme. In particular, the methanogenic SerRS relies on G1:C72 identity and on the number of unpaired nucleotides at the base of the variable stem for tRNASer recognition, unlike its bacterial type counterpart. We propose that such a distinction between the two enzymes in tRNASer identity determinants reflects their evolutionary pathways, hence attesting to their diversity. PMID- 15364940 TI - The role of thiol species in the hypertolerance of Aspergillus sp. P37 to arsenic. AB - Aspergillus sp. P37 is an arsenate-hypertolerant fungus isolated from a river in Spain with a long history of contamination with metals. This strain is able to grow in the presence of 0.2 M arsenate, i.e. 20-fold higher than the reference strain, Aspergillus nidulans TS1. Although Aspergillus sp. P37 reduces As(V) to As(III), which is slowly pumped out of the cell, the measured efflux of oxyanions is insufficient to explain the high tolerance levels of this strain. To gain an insight into this paradox, the accumulation of acid-soluble thiol species in Aspergillus sp. P37 when exposed to arsenic was compared with that of the arsenic sensitive A. nidulans TS1 strain. Increasing levels of arsenic in the medium did not diminish the intracellular pool of reduced glutathione in Aspergillus sp. P37, in sharp contrast with the decline of glutathione in A. nidulans under the same conditions. Furthermore, concentrations of arsenic that were inhibitory for the sensitive A. nidulans strain (e.g. 50 mM and above) provoked a massive formation of vacuoles filled with thiol species. Because the major fraction of the cellular arsenic was present as the glutathione conjugate As(GS)3, it is plausible that the arsenic-hypertolerant phenotype of Aspergillus sp. P37 is in part due to an enhanced capacity to maintain a large intracellular glutathione pool under conditions of arsenic exposure and to sequester As(GS)3 in vacuoles. High pressure liquid chromatography analysis of cell extracts revealed that the contact of Aspergillus sp. P37 (but not A. nidulans) with high arsenic concentrations (> or =150 mM) induced the production of small quantities of a distinct thiol species indistinguishable from plant phytochelatin-2. Yet, we argue that phytochelatins do not explain arsenic resistance in Aspergillus, and we advocate the role of As(GS)3 complexes in arsenic detoxification. PMID- 15364941 TI - Protein kinase C-related kinase 2 regulates hepatitis C virus RNA polymerase function by phosphorylation. AB - The hepatitis C virus (HCV) NS5B protein is the viral RNA-dependent RNA polymerase required for replication of the HCV RNA genome. We have identified a peptide that most closely resembles a short region of the protein kinase C related kinase 2 (PRK2) by screening of a random 12-mer peptide library displayed on the surface of the M13 bacteriophage with NS5B proteins immobilized on microwell plates. Competitive phage enzyme-linked immunosorbent assay with a synthetic peptide showed that the phage clone displaying this peptide could bind HCV RNA polymerase with a high affinity. Coimmunoprecipitation and colocalization studies demonstrated in vivo interaction of NS5B with PRK2. In vitro kinase assays demonstrated that PRK2 specifically phosphorylates NS5B by interaction with the N-terminal finger domain of NS5B (amino acids 1-187). Consistent with the in vitro NS5B-phosphorylating activity of PRK2, we detected the phosphorylated form of NS5B by metabolic cell labeling. Furthermore, HCV NS5B immunoprecipitated from HCV subgenomic replicon cells was specifically recognized by an antiphosphoserine antibody. Knock-down of the endogenous PRK2 expression using a PRK2-specific small interfering RNA inhibited HCV RNA replication. In contrast, PRK2 overexpression, which was accompanied by an increase of in the level of its active form, dramatically enhanced HCV RNA replication. Altogether, our results indicate that HCV RNA replication is regulated by NS5B phosphorylation by PRK2. PMID- 15364942 TI - Endogenous formation of novel halogenated 2'-deoxycytidine. Hypohalous acid mediated DNA modification at the site of inflammation. AB - A potential role of DNA damage by leukocyte-derived reactive species in carcinogenesis has been suggested. Leukocyte-derived peroxidases, such as myeloperoxidase and eosinophil peroxidase, use hydrogen peroxide and halides (Cl- and Br-) to generate hypohalous acids (HOCl and HOBr), halogenating intermediates. It has been suggested that these oxidants lead to the formation of halogenated products upon reaction with nucleobases. To verify the consequences of phagocyte-mediated DNA damage at the site of inflammation, we developed a novel monoclonal antibody (mAb2D3) that recognizes the hypohalous acid-modified DNA and found that the antibody most significantly recognized HOCl/HOBr-modified 2'-deoxycytidine residues. The immunoreactivity of HOCl-treated oligonucleotide was attenuated by excess methionine, suggesting that chloramine-like species may be the plausible epitopes of the antibody. On the basis of further characterization combined with mass spectrometric analysis, the epitopes of mAb2D3 were determined to be novel N4,5-dihalogenated 2'-deoxycytidine residues. The formation of the dihalogenated 2'-deoxycytidine in vivo was immunohistochemically demonstrated in the lung and liver nuclei of mice treated with lipopolysaccharides, an experimental inflammatory model. These results strongly suggest that phagocyte-derived oxidants, hypohalous acids, endogenously generate the halogenated DNA bases such as a novel dihalogenated 2'-deoxycytidine in vivo. Halogenation (chlorination and/or bromination) of DNA therefore may constitute one mechanism for oxidative DNA damage at the site of inflammation. PMID- 15364943 TI - Sugar recognition by the lactose permease of Escherichia coli. AB - Biochemical, luminescence and mass spectroscopy approaches indicate that Trp-151 (helix V) plays an important role in hydrophobic stacking with the galactopyranosyl ring of substrate and that Glu-269 (helix VIII) is essential for substrate affinity and specificity. The x-ray structure of the lactose permease (LacY) with bound substrate is consistent with these conclusions and suggests that a possible H-bond between Glu-269 and Trp-151 may play a critical role in the architecture of the binding site. We have now probed this relationship by exploiting the intrinsic luminescence of a single Trp-151 LacY with various replacements for Glu-269. Mutations at position 269 dramatically alter the environment of Trp-151 in a manner that correlates with binding affinity of LacY substrates. Furthermore, chemical modification of Trp-151 with N-bromosuccinimide indicates that Glu-269 forms an H-bond with the indole N. It is concluded that 1) an H-bond between the indole N and Glu-269 optimizes the formation of the substrate binding site in the inward facing conformation of LacY, and 2) the disposition of the residues implicated in sugar binding in different conformers suggests that sugar binding by LacY involves induced fit. PMID- 15364944 TI - Arginine methylation of scaffold attachment factor A by heterogeneous nuclear ribonucleoprotein particle-associated PRMT1. AB - Components of the heterogeneous nuclear ribonucleoprotein (hnRNP) complex and other nucleic acid-binding proteins are subject to methylation on specific arginine residues by the catalytic activity of arginine methyltransferases. The methylation has been implicated in transcriptional regulation and RNA and protein trafficking and signal transduction, but the mechanism by which these functions are achieved has remained undetermined. We show here that the predominant arginine methyltransferase in human cells, protein arginine methyltransferase 1 (PRMT1), is associated with hnRNP complexes, dependent on the methylation status of the cell, and that it methylates its preferred substrates in situ. Binding of PRMT1 occurs through physical interaction with scaffold attachment factor A (SAF A), also known as hnRNP-U, which is quantitatively methylated by PRMT1 in all investigated cell lines as determined by a novel, highly specific, methylation sensitive antibody. PMID- 15364945 TI - Release of beta-nicotinamide adenine dinucleotide upon stimulation of postganglionic nerve terminals in blood vessels and urinary bladder. AB - Chemical signaling in autonomic neuromuscular transmission involves agents that function as neurotransmitters and/or neuromodulators. Using high performance liquid chromatography techniques with fluorescence and electrochemical detection we observed that, in addition to ATP and norepinephrine (NE), electrical field stimulation (EFS, 4-16 Hz, 0.1-0.3 ms, 15 V, 60-120 s) of isolated vascular and non-vascular preparations co-releases a previously unidentified compound with apparent nucleotide or nucleoside structure. Extensive screening of more than 25 nucleotides and nucleosides followed by detailed peak identification revealed that beta-nicotinamide adenine dinucleotide (beta-NAD) is released in tissue superfusates upon EFS of canine mesenteric artery (CMA), canine urinary bladder, and murine urinary bladder in the amounts of 7.1 +/- 0.7, 26.5 +/- 4.5, and 15.1 +/- 3.2 fmol/mg of tissue, respectively. Smaller amounts of the beta-NAD metabolites cyclic adenosine 5'-diphosphoribose (cADPR) and ADPR were also present in the superfusates collected during EFS of CMA (2.5 +/- 0.9 and 5.8 +/- 0.8 fmol/mg of tissue, respectively), canine urinary bladder (1.8 +/- 0.5 and 9.0 +/- 6.0 fmol/mg of tissue, respectively), and murine urinary bladder (1.4 +/- 0.1 and 6.2 +/- 2.4 fmol/mg of tissue, respectively). The three nucleotides were also detected in the samples collected before EFS (0.2-1.6 fmol/mg of tissue). Exogenous beta-NAD, cADPR, and ADPR (all 100 nm) reduced the release of NE in CMA at 16 Hz from 27.8 +/- 6.0 fmol/mg of tissue to 15.5 +/- 5.0, 12 +/- 3.0, and 10.0 +/- 4.0 fmol/mg of tissue, respectively. In conclusion, we detected constitutive and nerve-evoked overflow of beta-NAD, cADPR, and ADPR in vascular and non-vascular smooth muscles, beta-NAD being the prevailing compound. These substances modulate the release of NE, implicating novel nucleotide mechanisms of autonomic nervous system control of smooth muscle. PMID- 15364946 TI - Cell-free reconstitution of transport from the trans-golgi network to the late endosome/prevacuolar compartment. AB - Vesicle-mediated transport between the trans-Golgi network (TGN) and the late endosome/prevacuolar compartment (PVC) is an essential step in lysosomal/vacuolar biogenesis. In addition, localization of integral membrane proteins to the TGN requires continual cycles of vesicular transport between the TGN and endosomal compartments. Genetic and biochemical analyses in yeast have identified a variety of proteins required for TGN-to-PVC transport. However, the precise mechanisms of vesicle formation, transport, and fusion have not been fully elucidated. To study the steps of TGN-to-PVC transport in mechanistic detail, we have developed a cell free assay to monitor delivery of the processing protease Kex2p from the TGN to PVC compartments containing a Kex2p substrate. Transport is time-, temperature-, and ATP-dependent and requires the t-SNARE Pep12p. Moreover, cell-free delivery of Kex2p to the PVC results in the co-integration of Kex2p into PVC membranes containing the Kex2p substrate as determined by co-immunoisolation of Kex2p and the substrate using antibody against the Kex2p cytosolic tail. This work represents the first cell-free reconstitution and biochemical analysis of the essential vacuolar/lysosomal sorting step TGN to late endosome transport. PMID- 15364947 TI - A new regulation of non-capacitative calcium entry in insect pacemaker neurosecretory neurons. Involvement of arachidonic acid, no-guanylyl cyclase/cGMP, and cAMP. AB - Efferent dorsal unpaired median neurons are pacemaker neurosecretory cells. A Ca(2+) background current contributing to the pacemaker activity of cockroach dorsal unpaired median neurons is up-regulated by neurohormone D (NHD), an octapeptide belonging to the adipokinetic hormone family. This modulation accelerates spiking and increases [Ca(2+)](i). Using patch clamp, calcium imaging, and immunocytochemistry, we investigated the signaling pathway of NHD induced current modulation. The membrane depolarization produced by NHD was related to the increase in membrane conductance for Ca(2+), Ba(2+), or Sr(2+). This increase was abolished by LOE 908, an inhibitor of noncapacitive Ca(2+) entry (NCCE), and it was strongly attenuated by the phospholipase C inhibitor U37122 and the diacylglycerol lipase inhibitor RHC80267. Arachidonic acid and ETYA mimicked the NHD effect on background current. This was abolished by l-NAME and ODQ, inhibitors of NO synthase and NO-sensitive guanylyl cyclase, respectively, but mimicked by the NO donor sodium nitroprusside and 8-bromo-cGMP. Immunocytochemistry using cGMP antibodies indicated that NHD and ETYA increase cGMP. Inhibition of protein kinase G with KT5823 and R(p)-8-pCPT-cGMPS had no effect, whereas zaprinast, a cGMP-specific phosphodiesterase 5,6,9 inhibitor, mimicked the NHD effect. Furthermore, inhibition of the cGMP-activated phosphodiesterase 2 by EHNA and trequinsin abolished the effect of NHD. We conclude that the final step of the NHD signal transduction is the phosphodiesterase 2-induced down-regulation of the cAMP level. This removes a depression of NCCE directly attributed to cAMP because inhibition of protein kinase A with KT5720, R(p)-cAMPS, and PKI14-22 amide did not mimic the NHD effect. We also demonstrate that any mechanism increasing the cGMP level can induce NCCE. PMID- 15364948 TI - Mitochondria-specific function of the dynamin family protein DLP1 is mediated by its C-terminal domains. AB - The dynamin superfamily of large GTPases has been implicated in a variety of distinct intracellular membrane remodeling events. One of these family members, DLP1/Drp1, is similar to conventional dynamins as it contains an N-terminal GTPase domain followed by a middle region (MID), an unconserved region (UC), and a coiled-coil (CC) domain. DLP1 has been shown to function in membrane-based processes distinct from conventional dynamin, most notably mitochondrial fission. In this study, we tested whether the functional specificities of DLP1 and dynamin stems from differences in the individual domains of these proteins by generating dynamin/DLP1 chimeras in which correlate domains had been interchanged. Here we report that three consecutive C-terminal domains of DLP1 (MID-UC-CC) contain information necessary for DLP1-specific function and removing any one of these domains results in a loss of DLP1 function. Importantly, the coiled-coil (CC) domain of DLP1 alone targets specifically and exclusively to mitochondria, implicating its involvement in localizing DLP1 to this organelle in vivo. The mitochondrial targeting information within the DLP1 CC domain is not sufficient to retarget dynamin to mitochondria but is still able to adequately function as an assembly domain in a dynamin background. These data suggest that whereas the GTPase domain of DLP1 provides an enzymatic function, other domains contain information for intermolecular assembly and mitochondrial targeting. PMID- 15364949 TI - Arrestin regulates MAPK activation and prevents NADPH oxidase-dependent death of cells expressing CXCR2. AB - Activation of CXCR2 IL-8 receptor leads to activation of extracellular signal regulated kinases 1 and 2 (ERK1/2) and rapid receptor endocytosis. Co immunoprecipitation and co-localization experiments showed that arrestin and CXCR2 form complexes with components of the ERK1/2 cascade following ligand stimulation. However, in contrast to the activation of the beta2-adrenergic receptor, arrestin was not necessary for ERK1/2 phosphorylation or receptor endocytosis. In contrast, beta-arrestin 1/2 double knockout cells showed greatly enhanced phosphorylation of ERK1/2, as well as phosphorylation of the stress kinases p38 and c-Jun N-terminal protein kinase. The stimulation of stress kinases in arrestin double knockout cells could be attenuated in the presence of diphenylene iodonium (DPI), an inhibitor of the NADPH oxidase, suggesting that reactive oxidant species (ROS) participated in mitogen-activated protein kinase (MAPK) activation. ROS could indeed be detected in IL-8-stimulated beta-arrestin 1/2 knockout cells, and cytoplasmic Rac was translocated to the membrane fraction, which is a prerequisite for oxidant formation. The oxidative burst induced cell death within 6 h of IL-8 stimulation of these cells, which could be prevented in the presence of DPI. These results indicate a novel function for arrestin, which is protection from an excessive oxidative burst, resulting from the sustained stimulation of G-protein-coupled receptors that cause Rac translocation. PMID- 15364950 TI - TreT, a novel trehalose glycosyltransferring synthase of the hyperthermophilic archaeon Thermococcus litoralis. AB - The gene cluster in Thermococcus litoralis encoding a multicomponent and binding protein-dependent ABC transporter for trehalose and maltose contains an open reading frame of unknown function. We cloned this gene (now called treT), expressed it in Escherichia coli, purified the encoded protein, and identified it as an enzyme forming trehalose and ADP from ADP-glucose and glucose. The enzyme can also use UDP- and GDP-glucose but with less efficiency. The reaction is reversible, and ADP-glucose plus glucose can also be formed from trehalose and ADP. The rate of reaction and the equilibrium favor the formation of trehalose. At 90 degrees C, the optimal temperature for the enzymatic reaction, the half maximal concentration of ADP-glucose at saturating glucose concentrations is 1.14 mm and the V(max) is 160 units/mg protein. In the reverse reaction, the half maximal concentration of trehalose at saturating ADP concentrations is 11.5 mm and the V(max) was estimated to be 17 units/mg protein. Under non-denaturating in vitro conditions the enzyme behaves as a dimer of identical subunits of 48 kDa. As the transporter encoded in the same gene cluster, TreT is induced by trehalose and maltose in the growth medium. PMID- 15364951 TI - Regulation of ADAM12 cell-surface expression by protein kinase C epsilon. AB - The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell surface proteins that have a major impact on cell behavior. These transmembrane anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin-like-, cysteine-rich, epidermal growth factor-like, and transmembrane domain; and a cytoplasmic tail. The 90-kDa mature form of human ADAM12 is generated in the trans-Golgi through cleavage of the prodomain by a furin-peptidase and is stored intracellularly until translocation to the cell surface as a constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell surface and that catalytic activity of PKCepsilon is required for this translocation. The following results support this conclusion: 1) treatment of cells with 0.1 microM phorbol 12-myristate 13-acetate (PMA) enhanced ADAM12 cell-surface immunostaining, 2) ADAM12 and PKCepsilon could be co immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCepsilon expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCepsilon mutant did not exhibit ADAM12 cell surface translocation upon PMA treatment. Finally, we demonstrate that the C1 and C2 domains of PKCepsilon both contain a binding site for ADAM12. These studies show that PKCepsilon plays a critical role in the regulation of ADAM12 cell surface expression. PMID- 15364952 TI - Identification of Tim40 that mediates protein sorting to the mitochondrial intermembrane space. AB - Most mitochondrial proteins are synthesized in the cytosol, imported into mitochondria, and sorted to one of the four mitochondrial subcompartments. Here we identified a new inner membrane protein, Tim40, that mediates sorting of small Tim proteins to the intermembrane space. Tim40 is essential for yeast cell growth, and its function in vivo requires six conserved Cys residues but not anchoring of the protein to the inner membrane by its N-terminal hydrophobic segment. Depletion of Tim40 impairs the import of small Tim proteins into mitochondria both in vivo and in vitro. In wild-type mitochondria, Tim40 forms a translocation intermediate with small Tim proteins prior to their assembly in the intermembrane space in vitro. These results suggest the essential role of Tim40 in sorting/assembly of small Tim proteins. PMID- 15364953 TI - In vivo cleavage of alpha2,6-sialyltransferase by Alzheimer beta-secretase. AB - beta-Site amyloid precursor protein-cleaving enzyme 1 (BACE1) is a membrane-bound aspartic protease that cleaves amyloid precursor protein to produce a neurotoxic peptide, Abeta, and is implicated in triggering the pathogenesis of Alzheimer disease. We previously reported that BACE1 cleaved rat beta-galactoside alpha2,6 sialyltransferase (ST6Gal I) that was overexpressed in COS cells and that the NH(2) terminus of ST6Gal I secreted from the cells (E41 form) was Glu(41). Here we report that BACE1 gene knock-out mice have one third as much plasma ST6Gal I as control mice, indicating that BACE1 is a major protease which is responsible for cleaving ST6Gal I in vivo. We also found that BACE1-transgenic mice have increased level of ST6Gal I in plasma. Secretion of ST6Gal I from the liver into the plasma is known to be up-regulated during the acute-phase response. To investigate the role of BACE1 in ST6Gal I secretion in vivo, we analyzed the levels of BACE1 mRNA in the liver, as well as the plasma levels of ST6Gal I, in a hepatopathological model, i.e. Long-Evans Cinnamon (LEC) rats. This rat is a mutant that spontaneously accumulates copper in the liver and incurs hepatic damage. LEC rats exhibited simultaneous increases in BACE1 mRNA in the liver and in the E41 form of the ST6Gal I protein, the BACE1 product, in plasma as early as 6 weeks of age, again suggesting that BACE1 cleaves ST6Gal I in vivo and controls the secretion of the E41 form. PMID- 15364954 TI - Identification of sialoadhesin as a dominant lymph node counter-receptor for mouse macrophage galactose-type C-type lectin 1. AB - In the sensitization phase of contact hypersensitivity in mice, dermal macrophages (MOs) expressing MO galactose-type C-type lectin1 (MGL1) are known to migrate from the dermis to lymph nodes (LNs) where they accumulate in the subcapsular sinus, interfollicular regions, and areas surrounding high endothelial venules. We hypothesize that the interactions between MGL1 and its ligands determine the localizations of MGL1-positive cells within the LNs. In the present study, our major aim was to isolate MGL1 counter-receptor(s) from lysates of LNs using affinity chromatography with immobilized recombinant MGL1. Fractions bound and eluted with EDTA were analyzed by SDS-PAGE and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. One of the predominant components was sialoadhesin (Sn, Siglec-1). Sn from lysates of LNs was immobilized on microtiter plates precoated with anti-Sn monoclonal antibody, and binding of recombinant MGL1 and adhesion of cells expressing MGL1 were tested. The binding of recombinant MGL1 to Sn was shown to be dependent on Ca2+ and N glycans on Sn. MGL1-transfected Chinese hamster ovary cells adhered to the Sn coated plates, whereas mock transfectants did not. Immunohistochemical localization of anti-Sn monoclonal antibody in LN coincided with the subcapsular sinus area to which recombinant MGL1 was bound. Furthermore, the distribution of MGL1+ cells after sensitization with FITC was demonstrated to overlap with that of Sn within the subcapsular sinus of draining LNs. These results suggest that Sn acts as an endogenous counter-receptor for MGL1. PMID- 15364955 TI - Molecular basis of anti-horseradish peroxidase staining in Caenorhabditis elegans. AB - Cross-reactivity with anti-horseradish peroxidase antiserum is a feature of many glycoproteins from plants and invertebrates; indeed staining with this reagent has been used to track neurons in Drosophila melanogaster and Caenorhabditis elegans. Although in insects the evidence indicates that the cross-reaction results from the presence of core alpha1,3-fucosylated N-glycans, the molecular basis for anti-horseradish peroxidase staining in nematodes has been unresolved to date. By using Western blots of wild-type and mutant C. elegans extracts in conjunction with specific inhibitors, we show that the cross-reaction is due to core alpha1,3-fucosylation. Of the various mutants examined, one with a deletion of the fut-1 (K08F8.3) gene showed no reaction to anti-horseradish peroxidase; the molecular phenotype was rescued by injection of either the K08F8 cosmid or the fut-1 open reading frame under control of the let-858 promoter. Furthermore, expression of fut-1 cDNA in Pichia and insect cells in conjunction with antibody staining, high pressure liquid chromatography, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analyses showed that FUT-1 is a core alpha1,3-fucosyltransferase with an unusual substrate specificity. It is the only core fucosyltransferase in plants and animals described to date that does not require the prior action of N-acetylglucosaminyltransferase I. PMID- 15364956 TI - LEC12 and LEC29 gain-of-function Chinese hamster ovary mutants reveal mechanisms for regulating VIM-2 antigen synthesis and E-selectin binding. AB - LEC12 and LEC29 are two gain-of-function Chinese hamster ovary glycosylation mutants that express the Fut9 gene encoding alpha(1,3)fucosyltransferase IX (alpha(1,3) Fuc-TIX). Both mutants express the Lewis X (Le(X)) determinant Galbeta(1,4)[Fucalpha(1,3)]GlcNAc, and LEC12, but not LEC29 cells, also express the VIM-2 antigen SAalpha(2,3) Galbeta(1,4)GlcNAcbeta(1,3)Galbeta(1,4)[Fucalpha(1,3)]GlcNAc. Here we show that LEC29 cells transfected with a Fut9 cDNA express VIM-2, and thus LEC29 cells synthesize appropriate acceptors to generate the VIM-2 epitope. Semiquantitative reverse transcription-PCR showed that LEC12 has 10- to 20-fold less Fut9 gene transcripts than LEC29. However, Western analysis revealed that LEC12 has approximately 20 times more Fut9 protein than LEC29. The latter finding was consistent with our previous observation that LEC12 has approximately 40 times more in vitro alpha(1,3)Fuc-T activity than LEC29. The basis for the difference in Fut9 protein levels was found to lie in sequence differences in the 5' untranslated regions (5'-UTR) of LEC12 and LEC29 Fut9 gene transcripts. Whereas reporter assays with the respective 5'-UTR regions linked to luciferase did not indicate a reduced translation efficiency caused by the LEC29 5'-UTR, transfected full-length LEC29 Fut9 cDNA or in vitro-synthesized full-length LEC29 Fut9 RNA gave less Fut9 protein than similar constructs with a LEC12 5'-UTR. This difference appears to be largely responsible for the reduced alpha(1,3)Fuc-TIX activity and lack of VIM-2 expression of LEC29 cells. This could be of physiological relevance, because LEC29 and parent Chinese hamster ovary cells transiently expressing a Fut9 cDNA were able to bind mouse E-selectin, although they did not express sialyl-Le(X). PMID- 15364957 TI - YidC family members are involved in the membrane insertion, lateral integration, folding, and assembly of membrane proteins. AB - Members of the YidC family exist in all three domains of life, where they control the assembly of a large variety of membrane protein complexes that function as transporters, energy devices, or sensor proteins. Recent studies in bacteria have shown that YidC functions on its own as a membrane protein insertase independent of the Sec protein-conducting channel. YidC can also assist in the lateral integration and folding of membrane proteins that insert into the membrane via the Sec pathway. PMID- 15364958 TI - Functional interaction between BLM helicase and 53BP1 in a Chk1-mediated pathway during S-phase arrest. AB - Bloom's syndrome is a rare autosomal recessive genetic disorder characterized by chromosomal aberrations, genetic instability, and cancer predisposition, all of which may be the result of abnormal signal transduction during DNA damage recognition. Here, we show that BLM is an intermediate responder to stalled DNA replication forks. BLM colocalized and physically interacted with the DNA damage response proteins 53BP1 and H2AX. Although BLM facilitated physical interaction between p53 and 53BP1, 53BP1 was required for efficient accumulation of both BLM and p53 at the sites of stalled replication. The accumulation of BLM/53BP1 foci and the physical interaction between them was independent of gamma-H2AX. The active Chk1 kinase was essential for both the accurate focal colocalization of 53BP1 with BLM and the consequent stabilization of BLM. Once the ATR/Chk1- and 53BP1-mediated signal from replicational stress is received, BLM functions in multiple downstream repair processes, thereby fulfilling its role as a caretaker tumor suppressor. PMID- 15364959 TI - Transcription-dependent spatial arrangements of CFTR and adjacent genes in human cell nuclei. AB - We investigated in different human cell types nuclear positioning and transcriptional regulation of the functionally unrelated genes GASZ, CFTR, and CORTBP2, mapping to adjacent loci on human chromosome 7q31. When inactive, GASZ, CFTR, and CORTBP2 preferentially associated with the nuclear periphery and with perinuclear heterochromatin, whereas in their actively transcribed states the gene loci preferentially associated with euchromatin in the nuclear interior. Adjacent genes associated simultaneously with these distinct chromatin fractions localizing at different nuclear regions, in accordance with their individual transcriptional regulation. Although the nuclear localization of CFTR changed after altering its transcription levels, the transcriptional status of CFTR was not changed by driving this gene into a different nuclear environment. This implied that the transcriptional activity affected the nuclear positioning, and not vice versa. Together, the results show that small chromosomal subregions can display highly flexible nuclear organizations that are regulated at the level of individual genes in a transcription-dependent manner. PMID- 15364960 TI - Mathematical modeling reveals threshold mechanism in CD95-induced apoptosis. AB - Mathematical modeling is required for understanding the complex behavior of large signal transduction networks. Previous attempts to model signal transduction pathways were often limited to small systems or based on qualitative data only. Here, we developed a mathematical modeling framework for understanding the complex signaling behavior of CD95(APO-1/Fas)-mediated apoptosis. Defects in the regulation of apoptosis result in serious diseases such as cancer, autoimmunity, and neurodegeneration. During the last decade many of the molecular mechanisms of apoptosis signaling have been examined and elucidated. A systemic understanding of apoptosis is, however, still missing. To address the complexity of apoptotic signaling we subdivided this system into subsystems of different information qualities. A new approach for sensitivity analysis within the mathematical model was key for the identification of critical system parameters and two essential system properties: modularity and robustness. Our model describes the regulation of apoptosis on a systems level and resolves the important question of a threshold mechanism for the regulation of apoptosis. PMID- 15364961 TI - Rb is required for progression through myogenic differentiation but not maintenance of terminal differentiation. AB - To investigate the requirement for pRb in myogenic differentiation, a floxed Rb allele was deleted either in proliferating myoblasts or after differentiation. Myf5-Cre mice, lacking pRb in myoblasts, died immediately at birth and exhibited high numbers of apoptotic nuclei and an almost complete absence of myofibers. In contrast, MCK-Cre mice, lacking pRb in differentiated fibers, were viable and exhibited a normal muscle phenotype and ability to regenerate. Induction of differentiation of Rb-deficient primary myoblasts resulted in high rates of apoptosis and a total inability to form multinucleated myotubes. Upon induction of differentiation, Rb-deficient myoblasts up-regulated myogenin, an immediate early marker of differentiation, but failed to down-regulate Pax7 and exhibited growth in low serum conditions. Primary myoblasts in which Rb was deleted after expression of differentiated MCK-Cre formed normal multinucleated myotubes that did not enter S-phase in response to serum stimulation. Therefore, Rb plays a crucial role in the switch from proliferation to differentiation rather than maintenance of the terminally differentiated state. PMID- 15364962 TI - Myotubes differentiate optimally on substrates with tissue-like stiffness: pathological implications for soft or stiff microenvironments. AB - Contractile myocytes provide a test of the hypothesis that cells sense their mechanical as well as molecular microenvironment, altering expression, organization, and/or morphology accordingly. Here, myoblasts were cultured on collagen strips attached to glass or polymer gels of varied elasticity. Subsequent fusion into myotubes occurs independent of substrate flexibility. However, myosin/actin striations emerge later only on gels with stiffness typical of normal muscle (passive Young's modulus, E approximately 12 kPa). On glass and much softer or stiffer gels, including gels emulating stiff dystrophic muscle, cells do not striate. In addition, myotubes grown on top of a compliant bottom layer of glass-attached myotubes (but not softer fibroblasts) will striate, whereas the bottom cells will only assemble stress fibers and vinculin-rich adhesions. Unlike sarcomere formation, adhesion strength increases monotonically versus substrate stiffness with strongest adhesion on glass. These findings have major implications for in vivo introduction of stem cells into diseased or damaged striated muscle of altered mechanical composition. PMID- 15364963 TI - Catch bonds govern adhesion through L-selectin at threshold shear. AB - Flow-enhanced cell adhesion is an unexplained phenomenon that might result from a transport-dependent increase in on-rates or a force-dependent decrease in off rates of adhesive bonds. L-selectin requires a threshold shear to support leukocyte rolling on P-selectin glycoprotein ligand-1 (PSGL-1) and other vascular ligands. Low forces decrease L-selectin-PSGL-1 off-rates (catch bonds), whereas higher forces increase off-rates (slip bonds). We determined that a force dependent decrease in off-rates dictated flow-enhanced rolling of L-selectin bearing microspheres or neutrophils on PSGL-1. Catch bonds enabled increasing force to convert short-lived tethers into longer-lived tethers, which decreased rolling velocities and increased the regularity of rolling steps as shear rose from the threshold to an optimal value. As shear increased above the optimum, transitions to slip bonds shortened tether lifetimes, which increased rolling velocities and decreased rolling regularity. Thus, force-dependent alterations of bond lifetimes govern L-selectin-dependent cell adhesion below and above the shear optimum. These findings establish the first biological function for catch bonds as a mechanism for flow-enhanced cell adhesion. PMID- 15364964 TI - Molecular prognostication for soft tissue sarcomas: are we ready yet? PMID- 15364965 TI - Can we now agree to use the same definition to measure response according to CA 125? PMID- 15364966 TI - Should CA-125 response criteria be preferred to response evaluation criteria in solid tumors (RECIST) for prognostication during second-line chemotherapy of ovarian carcinoma? AB - PURPOSE: The aim of the study was to compare the prognostic value of a response by the Gynecologic Cancer Intergroup (GCIG) Cancer Antigen (CA) -125 response criteria and the Response Evaluation Criteria in Solid Tumors (RECIST) on survival in patients with ovarian carcinoma receiving second-line chemotherapy. PATIENTS AND METHODS: From a single-institution registry of 527 consecutive patients with primary ovarian carcinoma, 131 records satisfied the inclusion criteria: ovarian carcinoma of International Federation of Gynecology and Obstetrics stage IC to IV, first-line chemotherapy with paclitaxel and a platinum compound, refractory or recurrent disease, and second-line chemotherapy consisting of topotecan or paclitaxel plus carboplatin. Univariate and multivariate analyses of survival were performed using the landmark method. RESULTS: In patients with measurable disease by RECIST and with assessable disease by the CA-125 criteria (n = 68), the CA-125 criteria were 2.6 times better than the RECIST at disclosing survival. In a multivariate Cox analysis with inclusion of nine potential prognostic parameters, CA-125 response (responders v nonresponders; hazard ratio, 0.21; P < .001) and number of relapse sites (solitary v multiple; hazard ratio, 0.47; P = .020) were identified as contributory prognostic factors for survival, whereas the parameters of RECIST (responders v nonresponders), as well as the remaining variables, had nonsignificant prognostic impact. CONCLUSION: The GCIG CA-125 response criteria are a better prognostic tool than RECIST in second-line treatment with topotecan or paclitaxel plus carboplatin in patients with ovarian carcinoma. PMID- 15364967 TI - Histologic grade, but not SYT-SSX fusion type, is an important prognostic factor in patients with synovial sarcoma: a multicenter, retrospective analysis. AB - PURPOSE: To assess the prognostic value of SYT-SSX fusion type, in comparison with other factors, in a population of 165 patients with synovial sarcoma (SS). PATIENTS AND METHODS: Data on 165 patients with SS (141 with localized disease at diagnosis) were studied retrospectively. The following parameters were examined for their potential prognostic value: age at diagnosis, sex, tumor site (extremities v proximal/truncal), size, histology, mitotic count, necrosis, histologic grade (Federation Nationale des Centres de Lutte Contre le Cancer system), stage (1997 tumor-node-metastasis system classification), surgical margin status (assessed histologically), and fusion type (SYT-SSX1 v SYT-SSX2). Median follow-up time was 37 months (range, 2 to 302 months). RESULTS: Among those patients with localized disease at diagnosis, median and 5-year disease specific survivals (DSS) for the SYT-SSX1 and SYT-SSX2 subgroups were 126 months and 67.4% versus 82 months and 63.2%, respectively (P = .12). Median and 5-year metastasis-free survivals (MFS) were 84 months and 54.2% for SYT-SSX1 versus 50 months and 47.6% for SYT-SSX2 (P = .76). Univariate analyses showed that high histologic grade (grade 3), high mitotic count (>/= 10 mitoses/10 high-power fields), stage III disease, size greater than 7 cm, tumor necrosis, and presence of areas of poorly differentiated morphology were significant adverse prognostic factors for DSS and MFS, whereas SYT-SSX fusion type, tumor histology (biphasic v monophasic), and patient sex were not. Age greater than 35 years adversely affected DSS but not MFS. In multivariate analyses, histologic grade was the most significant prognostic factor for both DSS and MFS. CONCLUSION: For patients with localized SS, histologic grade but not SYT-SSX fusion type is a strong predictor of survival. PMID- 15364968 TI - GB virus type C: a beneficial infection? PMID- 15364969 TI - Detection of intrahepatic hepatitis B virus DNA and correlation with hepatic necroinflammation and fibrosis. AB - Assessment of intrahepatic hepatitis B virus (HBV) DNA levels in patients with chronic hepatitis B is important in understanding the natural history of the disease and designing antiviral therapy regimens. However, there is no standardized method for the measurement of intrahepatic HBV DNA levels. We describe a convenient novel method for the measurement of intrahepatic HBV DNA levels based on a modified COBAS Amplicor HBV Monitor test for HBV DNA measurement and real-time PCR beta-actin gene detection for human genomic DNA (hgDNA) quantitation. Fifteen hepatitis B e antigen (HBeAg)-positive patients, 26 patients positive for antibody to HBeAg (anti-HBe), and 8 control patients were recruited. The mean between-run coefficient of variation for the beta-actin real time PCR assay was 15.4%. All eight control patients had undetectable intrahepatic and serum HBV DNA levels. All chronic hepatitis B patients had detectable intrahepatic HBV DNA levels, and all but one anti-HBe-positive patient had detectable serum HBV DNA levels. HBeAg-positive patients had higher median intrahepatic and serum HBV DNA levels than anti-HBe-positive patients (6,950 versus 676 HBV DNA copies/ng of hgDNA, respectively [P < 0.001] and 184 x 10(6) versus 6.65 x 10(6) copies/ml, respectively [P < 0.001]). The intrahepatic HBV DNA levels correlated strongly with the serum HBV DNA levels (r = 0.842; P < 0.001) and with the degree of fibrosis (P = 0.014). We conclude that the method that we describe is reliable and convenient for the measurement of intrahepatic HBV DNA levels and has potential clinical significance. PMID- 15364970 TI - rpoB gene sequencing for identification of Corynebacterium species. AB - The genus Corynebacterium is a heterogeneous group of species comprising human and animal pathogens and environmental bacteria. It is defined on the basis of several phenotypic characters and the results of DNA-DNA relatedness and, more recently, 16S rRNA gene sequencing. However, the 16S rRNA gene is not polymorphic enough to ensure reliable phylogenetic studies and needs to be completely sequenced for accurate identification. The almost complete rpoB sequences of 56 Corynebacterium species were determined by both PCR and genome walking methods. In all cases the percent similarities between different species were lower than those observed by 16S rRNA gene sequencing, even for those species with degrees of high similarity. Several clusters supported by high bootstrap values were identified. In order to propose a method for strain identification which does not require sequencing of the complete rpoB sequence (approximately 3,500 bp), we identified an area with a high degree of polymorphism, bordered by conserved sequences that can be used as universal primers for PCR amplification and sequencing. The sequence of this fragment (434 to 452 bp) allows accurate species identification and may be used in the future for routine sequence-based identification of Corynebacterium species. PMID- 15364971 TI - Long-term follow-up study of Chinese patients with YMDD mutations: significance of hepatitis B virus genotypes and characteristics of biochemical flares. AB - We sought to examine the role of hepatitis B virus (HBV) genotypes in virological breakthroughs and biochemical flares in patients with YMDD mutations during lamivudine therapy. Virologic breakthroughs (i.e., the reappearance of HBV DNA as determined by bDNA assay) and biochemical flares (mild flares = alanine aminotransferase [ALT] between 2 and 10 times the upper limit of normal [ULN]; severe flares = ALT >10 times ULN) were monitored in 154 hepatitis B e antigen positive patients receiving long-term lamivudine. The HBV genotypes and YMDD mutations were determined. Forty-three patients had virological breakthroughs with YMDD mutations (median follow-up of 29.6 months [range, 22.3 to 61.4]). Twenty patients (47%) patients had mild biochemical flares; seven (16%) had severe flares. Two patients showed an elevation of bilirubin level that is >2 times the ULN. All patients recovered spontaneously. The cumulative risks for biochemical flares were 28, 47, and 58% for the first 3 years, respectively. Patients with biochemical flares compared to those without flares had a significantly higher median pretreatment ALT level (61 U/liter versus 34.5 U/liter [P = 0.012]). There were no differences in the cumulative risk of virological breakthroughs, risk, and severity of biochemical flares between patients with genotypes B (n = 11) and C (n = 32). There was an increase in the percentage of patients with single YMDD mutant at last follow-up compared to that at the time of virological breakthroughs (74% [n = 32] versus 47% [n = 20], respectively; P = 0.015). The chances of YMDD mutations with virological breakthroughs and biochemical flares were similar in patients with genotypes B and C. Biochemical flares were common, with 16% being severe in nature. High pretreatment ALT levels were associated with a higher chance of biochemical flares. PMID- 15364972 TI - Is the perceived association between Chlamydia pneumoniae and vascular diseases biased by methodology? AB - Inter- and intralaboratory inconsistencies in detection rates of Chlamydia pneumoniae in vascular specimens have been demonstrated. In this study, 66 vascular tissue specimens from 66 patients with vascular disease were tested by three PCR assays: a 16S PCR-based reverse line blot (RLB) assay, a single-step PCR, and a nested PCR. Also, we explored the impacts of different DNA polymerase enzymes on the results based on gel electrophoresis and hybridization. The PCR results by gel electrophoresis in the single-step PCR depended on which DNA polymerase was used. All samples were negative with AmpliTaq Gold DNA polymerase, and 54.5% (36 of 66) were positive with the conventional Taq DNA polymerase. All samples were negative after hybridization with a C. pneumoniae-specific probe. In the nested PCR, all specimens were negative by gel electrophoresis and after hybridization. The RLB assay failed to detect C. pneumoniae in any specimen; however, 20 specimens were Chlamydia sp. positive. The sequence analysis of six of these samples demonstrated Chlamydia-like organisms. RLB detected Chlamydia sp. DNA in water and in the elution buffer after passage of the Qiagen columns (11 of 40). This study identified factors that may influence the detection of C. pneumoniae DNA in vascular tissues and consequently bias the perception of a link between C. pneumoniae and vascular diseases. The following are strongly recommended: to use DNA polymerases that have to be activated, to decontaminate with dUTP-uracil-DNA glycosylase, to hybridize with specific probes, to include sufficient controls, and to use molecular grade water. PMID- 15364973 TI - Characteristics of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis isolated from the nasopharynges of asymptomatic children and molecular analysis of S. pneumoniae and H. influenzae strain replacement in the nasopharynx. AB - Nasopharyngeal carriage of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in 226 children in different settings (in a creche [day care center], in an orphanage, and at home) during two seasons (winter and spring) was studied. The rates of carriage of S. pneumoniae and H. influenzae were markedly higher in the creche and in the orphanage than in the home setting (e.g., 56.5, 63.3, and 25.9%, respectively, for S. pneumoniae in winter). Approximately 80% of the S. pneumoniae isolates identified in the creche and in the orphanage belonged to the serotypes represented in the seven-valent pneumococcal vaccine, and 4.4% of the children were colonized by H. influenzae type b. Almost all H. influenzae isolates were fully susceptible to the antimicrobial agents tested, and only five (3.6%) produced beta-lactamase; in contrast, 100% of the M. catarrhalis isolates were beta-lactamase positive. Among S. pneumoniae isolates, 36.2% were nonsusceptible to penicillin (PNSP) and 11.8% were fully resistant to penicillin (PRP). All PNSP isolates were obtained from children at the creche and at the orphanage but not among children brought up at home, and all PRP isolates showed a multiresistant phenotype. Colonization by PRP isolates correlated well with prior treatment with beta-lactams. For the majority of children colonized at both sampling times, strain replacement of S. pneumoniae and H. influenzae was observed; long-term colonization by a single strain was rare. PMID- 15364974 TI - Mutation in a Lordsdale norovirus epidemic strain as a potential indicator of transmission routes. AB - An increase in norovirus outbreaks was reported internationally during 2002 and 2003 and was also observed in Oxfordshire (United Kingdom) hospitals. To understand their epidemiological relationships, viruses from 22 outbreaks (15 from one hospital) were subjected to nucleotide sequencing. The 3'-terminal 3,255 nt or complete genomes were determined for 49 viruses. All outbreaks were caused by a genogroup II norovirus related to the Lordsdale virus (GII 4), common in healthcare settings. The norovirus mutation rate was sufficiently high that the 3,255-nucleotide sequences allowed separate and potentially connected outbreaks to be identified, since all outbreaks with identical sequences were temporally or geographically linked. The high mutation rate was further indicated by four mutations and three microheterogeneities in 3,255 nucleotides during 17 days of norovirus shedding by an immunocompromised patient. The data suggested that multiple virus introductions from the community, occasional transmission among wards, and one instance of ongoing environmental contamination had occurred. The accumulation, or lack, of mutations within an outbreak was also used to indicate the predominant transmission route. In an outbreak where person-to-person spread was thought to predominate, six mutations were detected throughout the genome, whereas one mutation was detected when point source infection was suspected. This norovirus epidemic strain differed from its closest previously described relative by 11.4 to 13.6% in the outer P2 domain of the capsid, which also had a single amino-acid insertion. Alterations to the capsid structure compared to previous noroviruses may explain the increased number of outbreaks during 2002 and 2003. PMID- 15364975 TI - The species Mycobacterium africanum in the light of new molecular markers. AB - The findings of recent studies addressing the molecular characteristics of Mycobacterium tuberculosis complex isolates have initiated a discussion on the classification of M. africanum, especially of those isolates originating from East Africa (cluster F, subtype II) and displaying phenotypic and biochemical characteristics more similar to those of M. tuberculosis. To further address this question, we analyzed a representative collection of 63 M. tuberculosis complex strains comprising 30 M. africanum subtype I strains, 20 M. africanum subtype II strains, 10 randomly chosen M. tuberculosis isolates, and type strains of M. tuberculosis, M. bovis, and M. africanum for the following biochemical and molecular characteristics: single-nucleotide polymorphisms (SNPs) in gyrB and narGHJI and the presence or absence of RD1, RD9, and RD12. For all molecular markers analyzed, subtype II strains were identical to the M. tuberculosis strains tested. In contrast, the subtype I strains as well as the M. africanum type strain showed unique combinations of SNPs in gyrB and genomic deletions (the absence of RD9 and the presence of RD12), which proves their independence from M. tuberculosis and M. bovis. Accordingly, all subtype I strains displayed main biochemical characteristics included in the original species description of M. africanum. We conclude that the isolates from West Africa were proved to be M. africanum with respect to the phenotypic and genetic markers analyzed, while the isolates from East Africa must be regarded as phenotypic variants of M. tuberculosis (genotype Uganda). We propose the addition of the molecular characteristics defined here to the species description of M. africanum, which will allow clearer species differentiation in the future. PMID- 15364976 TI - Comprehensive detection and serotyping of human adenoviruses by PCR and sequencing. AB - Human adenoviruses are common pathogens associated with many diseases, including respiratory, gastrointestinal, and ocular infections. Because they are now being increasingly recognized as agents of life-threatening disseminated infection in immunocompromised patients, robust and sensitive laboratory detection methods are needed for their rapid diagnosis. We describe here a PCR assay using a single primer pair, targeting a region of the hexon gene containing hypervariable region 7, that can detect all known human adenovirus serotypes and allows for serotype determination through the analysis of the nucleotide sequence. This comprehensive assay has proven effective for diagnosing adenoviruses at the serotype level in a broad range of patient specimens, including conjunctival, nasopharyngeal, stool, blood, and urine specimens. PMID- 15364977 TI - Identification of emerging human-pathogenic Pythium insidiosum by serological and molecular assay-based methods. AB - Pythium insidiosum is a pathogen that causes disease in both animals and humans. Human infection is rare; however, when it does occur, most patients, especially those having underlying hemoglobinopathy syndromes, such as thalassemia, exhibit a severe form. We identified four isolates of P. insidiosum. Two were recovered from tissue biopsy specimens from thalassemic and leukemic patients, one was derived from brain tissue from a thalassemic patient, and another was isolated from a corneal ulcer from a fourth patient. Western blotting and an enzyme-linked immunosorbent assay (ELISA) were performed with a serum sample derived from one thalassemic patient. The methods used to identify the P. insidiosum isolates were based on morphology, nucleic acid sequencing, and a PCR assay. To confirm the identification, portions of the 18S rRNA genes of these four isolates were sequenced. The sequences were shown to be homologous to previously described P. insidiosum DNA sequences. In addition, PCR amplification of the internal transcribed spacer region specific for P. insidiosum was positive for all four isolates. The ELISA with the serum sample from the thalassemic patient gave a positive result from a serum dilution of 1:800. Finally, Western immunoblotting with this serum sample showed positive immunoglobulin G recognition for proteins of 110, 73, 56, 42 to 35, 30 to 28, 26, and 23 kDa. The results of this study show that both molecularly based diagnostic and serodiagnostic techniques are useful for the rapid identification of human pythiosis. The predominant antigens recognized by Western blotting may be useful in the development of a more sensitive and specific diagnostic tool for this disease. PMID- 15364978 TI - Comparison of two culture media and three sampling techniques for sensitive and rapid screening of vaginal colonization by group B streptococcus in pregnant women. AB - The Centers for Disease Control and Prevention (CDC) recommend universal screening of all pregnant women between 35 and 37 weeks of gestation for group B streptococci (GBS) by use of a selective broth medium. Recent reports suggest that Granada medium can be used for rapid and direct visual identification of GBS colonies. However, studies comparing the Granada medium method to the selective broth method are few, and while some report comparable sensitivities, others have found significant differences in detection rates between the two methods. This prospective study compared a method using Granada agar to a Todd-Hewitt broth method with subculture to blood agar in order to determine which GBS detection method is more sensitive and less labor-intensive and has a more rapid turnaround time. Detection rates for three sampling techniques (rectovaginal, vaginal only, and cervical only) were also compared. Consecutive specimens for GBS screening received over a 6-month period from 1,635 pregnant women were included. Overall, GBS was detected in 390 (23.8%) women. The Granada medium gave positive results for 348 of these women, and the selective broth gave positive results for 385, indicating sensitivities of 89.2% for the Granada medium and 98.7% for the selective broth. These findings show that the Granada medium method is less sensitive than the selective broth method and should not replace it as the only method for screening pregnant women for GBS. However, the Granada medium method reduced detection time to 1 day and also reduced the use of ancillary tests in approximately 90% of positive cases. Additionally, no significant differences were noted in the detection rates with rectovaginal, vaginal, and cervical specimens. PMID- 15364979 TI - Nosocomial transmission of CTX-M-2 beta-lactamase-producing Acinetobacter baumannii in a neurosurgery ward. AB - Three strains of cefotaxime (CTX)-resistant Acinetobacter baumannii, FM0209680, FM0300106, and FM0301433, were isolated from transtracheal aspirate cultures of three patients with probable nosocomial infections in a neurosurgery ward in Japan. The CTX MICs for these isolates were greater than 128 microg/ml but were drastically reduced in the presence of 4 microg of clavulanic acid per ml. These strains were also resistant to ceftriaxone, cefpodoxime, and aztreonam but were susceptible to ceftazidime and imipenem. The profile of resistance to various broad-spectrum beta-lactams was transferred by conjugation. Strain FM0209680 was not eradicated from case patient 1 by administration of imipenem, ceftazidime, and levofloxacin, even after a 6-month hospitalization period. Strains FM0300106 and FM0301433 were isolated from case patients 2 and 3 during the sixth week following admission, respectively, and then each patient was colonized for 3 weeks. Eradication of FM0300106 was successfully obtained from case patient 2 by imipenem treatment, while administration of imipenem was continued to prevent pneumonia. Prophylactic antimicrobial therapy was discontinued in case patient 3 because of the lack of pneumonic symptoms, and FM0301433 disappeared after the discontinuation of antimicrobial chemotherapy. All three strains carried the bla(CTX-M-2) gene, and the appearance of colonies in the growth-inhibitory zones around disks of CTX and aztreonam in double-disk synergy tests suggested inducible beta-lactamase production in these A. baumannii strains. The ribotyping investigation suggested that all these strains belong to the same clonal lineage. The plasmids harbored by A. baumannii had the same restriction profile as those harbored by Proteus mirabilis strains previously isolated in a urology ward of the Funabashi Medical Center. PMID- 15364980 TI - Use of culture, PCR analysis, and DNA microarrays for detection of Campylobacter jejuni and Campylobacter coli from chicken feces. AB - A DNA microarray for detection of Campylobacter spp. was recently developed and applied to detect Campylobacter spp. directly from chicken feces. Sixty-five pooled chicken cloacal swab samples from 650 individual broiler chickens were included in the study. The results of Campylobacter sp. detection obtained with DNA microarrays were compared to those obtained by conventional culture and gel electrophoresis. By conventional culture, 60% of the samples were positive for either Campylobacter jejuni or Campylobacter coli. By PCR and capillary electrophoresis, 95% of the samples were positive for Campylobacter spp., whereas with DNA microarrays all samples were positive for Campylobacter spp. By application of DNA microarray analysis, the isolates in 4 samples (6%) could not be identified to the species level, whereas by PCR-capillary electrophoresis, the isolates in 12 samples (19%) remained unidentified. Interestingly, PCR-capillary electrophoresis analysis revealed that two (3%) of the samples were positive for both C. jejuni and C. coli, while DNA microarray analysis revealed that nine (14%) of the samples were positive for both species. Of 65 samples, 2 samples were identified to contain C. coli by conventional culture but were positive for C. jejuni by both PCR-capillary electrophoresis and DNA microarray analysis. The discrepancy between the methods is discussed. PMID- 15364981 TI - Enumeration and isolation of cpe-positive Clostridium perfringens spores from feces. AB - A hydrophobic grid membrane filter-colony hybridization (HGMF-CH) method for the enumeration and isolation of cpe gene-carrying (cpe-positive) Clostridium perfringens spores from feces was developed. A 425-bp DNA probe specific for the cpe gene was sensitive and specific when tested with bacterial DNA and pure cultures. The enumeration of cpe-positive C. perfringens by the HGMF-CH method proved to be as sensitive as nested PCR combined with the most-probable number technique when tested with fecal samples from healthy individuals. With the aid of the HGMF-CH method, positive hybridization signals were detected from two out of seven fecal samples obtained from healthy individuals. Furthermore, cpe positive C. perfringens was successfully isolated from both of these samples. The detection of cpe-positive C. perfringens by the HGMF-CH method is dependent on the ratio of cpe-positive C. perfringens colonies to total C. perfringens colonies growing on the HGMF-tryptose-sulfite-cycloserine plate. cpe-positive C. perfringens could be isolated if the ratio of cpe-positive C. perfringens spores to total C. perfringens spores was 6 x 10(-5) or higher. The HGMF-CH method provides an aid in the investigation of fecal samples of patients suffering from food poisoning or other diseases caused by cpe-positive C. perfringens. The method also offers a new approach in the investigation of the epidemiology of cpe positive C. perfringens strains. PMID- 15364982 TI - Epidemiology and molecular characterization of Streptococcus pyogenes recovered from scarlet fever patients in central Taiwan from 1996 to 1999. AB - One hundred seventy-nine Streptococcus pyogenes isolates recovered from scarlet fever patients from 1996 to 1999 in central Taiwan were characterized by emm, Vir, and pulsed-field gel electrophoresis (PFGE) typing methods. The protocols for Vir and PFGE typing were standardized. A database of the DNA fingerprints for the isolates was established. Nine emm or emm-like genes, 19 Vir patterns, and 26 SmaI PFGE patterns were detected among the isolates. Among the three typing methods, PFGE was the most discriminatory. However, it could not completely replace Vir typing because some isolates with identical PFGE patterns could be further differentiated into several Vir patterns. The prevalent emm types were emm4 (n = 81 isolates [45%]), emm12 (n = 64 [36%]), emm1 (n = 14 [8%]), and emm22 (n = 13 [7%]). Some emm type isolates could be further differentiated into several emm-Vir-PFGE genotypes; however, only one genotype in each emm group was usually predominant. DNA from nine isolates was resistant to SmaI digestion. Further PFGE analysis with SgrAI showed that the SmaI digestion-resistant strains could be derived from indigenous strains by horizontal transfer of exogenous genetic material. The emergence of the new strains could have resulted in an increase in scarlet fever cases in central Taiwan since 2000. The emm sequences, Vir, and PFGE pattern database will serve as a basis for information for the long term evolutionary study of local S. pyogenes strains. PMID- 15364983 TI - Phage types and genotypes of shiga toxin-producing Escherichia coli O157:H7 isolates from humans and animals in spain: identification and characterization of two predominating phage types (PT2 and PT8). AB - Phage typing and DNA macrorestriction fragment analysis by pulsed-field electrophoresis (PFGE) were used for the epidemiological subtyping of a collection of Shiga toxin-producing Escherichia coli (STEC) O157:H7 strains isolated in Spain between 1980 and 1999. Phage typing distinguished a total of 18 phage types among 171 strains isolated from different sources (67 humans, 82 bovines, 12 ovines, and 10 beef products). However, five phage types, phage type 2 (PT2; 42 strains), PT8 (33 strains), PT14 (14 strains), PT21/28 (11 strains), and PT54 (16 strains), accounted for 68% of the study isolates. PT2 and PT8 were the most frequently found among strains from both humans (51%) and bovines (46%). Interestingly, we detected a significant association between PT2 and PT14 and the presence of acute pathologies. A group of 108 of the 171 strains were analyzed by PFGE, and 53 distinct XbaI macrorestriction patterns were identified, with 38 strains exhibiting unique PFGE patterns. In contrast, phage typing identified 15 different phage types. A total of 66 phage type-PFGE subtype combinations were identified among the 108 strains. PFGE subtyping differentiated between unrelated strains that exhibited the same phage type. The most common phage type-PFGE pattern combinations were PT2-PFGE type 1 (1 human and 11 bovine strains), PT8 PFGE type 8 (2 human, 6 bovine, and 1 beef product strains), PT2-PFGE subtype 4A (1 human, 3 bovine, and 1 beef product strains). Nine (29%) of 31 human strains showed phage type-PFGE pattern combinations that were detected among the bovine strains included in this study, and 26 (38%) of 68 bovine strains produced phage type-PFGE pattern combinations observed among human strains included in this study, confirming that cattle are a major reservoir of strains pathogenic for humans. PT2 and PT8 strains formed two groups which differed from each other in their motilities, stx genotypes, PFGE patterns, and the severity of the illnesses that they caused. PMID- 15364984 TI - Identification to the species level and differentiation between strains of Aspergillus clinical isolates by automated repetitive-sequence-based PCR. AB - A commercially available repetitive-sequence-based PCR (rep-PCR) DNA fingerprinting assay adapted to an automated format, the DiversiLab system, enables rapid microbial identification and strain typing. We explored the performance of the DiversiLab system as a molecular typing tool for 69 Aspergillus isolates (38 A. fumigatus, 15 A. flavus, and 16 A. terreus isolates) had been previously characterized by morphological analysis. Initially, 27 Aspergillus isolates (10 A. fumigatus, 9 A. flavus, and 8 A. terreus isolates) were used as controls to create a rep-PCR-based DNA fingerprint library with the DiversiLab software. Then, 42 blinded Aspergillus isolates were typed using the system. The rep-PCR-based profile revealed 98% concordance with morphology-based identification. rep-PCR-based DNA fingerprints were reproducible and were consistent for DNA from both hyphae and conidia. DiversiLab dendrogram reports correctly identified all A. fumigatus (n = 28), A. terreus (n = 8), and A. flavus (n = 6) isolates in the 42 blinded Aspergillus isolates. rep-PCR-based identification of all isolates was 100% in agreement with the contiguous internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) sequence-based identification of the respective isolates. Additionally, the DiversiLab system could demonstrate strain-level differentiation of A. flavus and A. terreus. Automated rep-PCR may be a time-efficient, effective, easy-to-use, novel genotyping tool for identifying and determining the strain relatedness of fungi. This system may be useful for epidemiological studies, molecular typing, and surveillance of Aspergillus species. PMID- 15364985 TI - Microevolution of Candida albicans strains during catheter-related candidemia. AB - We examined microevolution in a series of Candida albicans strains isolated from patients with catheter-related candidemia. Sixty-one isolates (29 from blood, 18 from catheters, 10 from urine, and 4 from other sites) were obtained from 15 patients who were admitted to the same hospital over a 3-year period. Isolates were analyzed by using Southern hybridization with the C1 fragment of Ca3 as a probe (C1 fingerprinting) and pulsed-field gel electrophoresis (PFGE). PFGE typing consisted of electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG) by using SfiI (REAG-S) and BssHII (REAG-B). When catheter isolates were compared with blood isolates from the same patient, catheter isolates from 5 of 14 patients (36%) exhibited minor band differences (microevolution) relative to blood isolates in either C1 fingerprinting (n = 4), REAG-S (n = 3), or REAG-B (n = 5) profiles, although they had identical EK patterns. However, the other sequential isolates from each patient, which had identical EK patterns, showed the same REAG and C1 fingerprinting patterns. Both fingerprinting methods revealed that two distinct genotypes were shared by isolates from seven patients in a neonatal intensive care unit, suggesting two nosocomial clusters. Except for two catheter isolates from the index patients of each cluster, no consecutive isolates collected from each of the two clusters showed any microevolution during the 2- or 7-month cluster periods. The findings suggest that in catheter-related candidemia, some C. albicans strains undergo microevolution during catheter colonization. PMID- 15364986 TI - Characterizations of adenovirus type 41 isolates from children with acute gastroenteritis in Japan, Vietnam, and Korea. AB - Genetic and antigenic characterizations of 70 strains of adenovirus type 41 (Ad41), isolated between 1998 and 2001 from children in Japan, Vietnam, and Korea, were done by DNA restriction enzyme (RE) analysis, sequencing analysis, and monoclonal antibody (MAb)-based enzyme-linked immunosorbent assay (ELISA). Eight genome types were observed in the present study, among which D25, D26, D27, and D28 were novel genome types. These eight genome types were divided into two genome-type clusters (GTCs) based on phylogenetic analysis of the hypervariable regions (HVRs) of the hexon. GTC1 includes D1, D25, D26, D27, and D28, and the GTC2 contains D4, D12, and D22. The amino acid homologies among the members within a GTC were 97 to 100%, whereas between the members of different GTCs the homologies were 92 to 94%. The specificity of the GTC classification was confirmed by ELISA with MAb 1F, which was selected by the Ad41 prototype Tak strain. It was found that only the isolates of GTC1 but not of GTC2 reacted with MAb 1F. These results suggest that Ad41 isolates from the three countries should be classified into two subtypes. The accumulation of amino acid mutations located in HVRs of hexon are indicative for the classification of Ad41 subtype. PMID- 15364987 TI - Definition of the Beijing/W lineage of Mycobacterium tuberculosis on the basis of genetic markers. AB - Mycobacterium tuberculosis Beijing genotype strains are highly prevalent in Asian countries and in the territory of the former Soviet Union. They are increasingly reported in other areas of the world and are frequently associated with tuberculosis outbreaks and drug resistance. Beijing genotype strains, including W strains, have been characterized by their highly similar multicopy IS6110 restriction fragment length polymorphism (RFLP) patterns, deletion of spacers 1 to 34 in the direct repeat region (Beijing spoligotype), and insertion of IS6110 in the genomic dnaA-dnaN locus. In this study the suitability and comparability of these three genetic markers to identify members of the Beijing lineage were evaluated. In a well-characterized collection of 1,020 M. tuberculosis isolates representative of the IS6110 RFLP genotypes found in The Netherlands, strains of two clades had spoligotypes characteristic of the Beijing lineage. A set of 19 Beijing reference RFLP patterns was selected to retrieve all Beijing strains from the Dutch database. These reference patterns gave a sensitivity of 98.1% and a specificity of 99.7% for identifying Beijing strains (defined by spoligotyping) in an international database of 1,084 strains. The usefulness of the reference patterns was also assessed with large DNA fingerprint databases in two other European countries and for identification strains from the W lineage found in the United States. A standardized definition for the identification of M. tuberculosis strains belonging to the Beijing/W lineage, as described in this work, will facilitate further studies on the spread and characterization of this widespread genotype family of M. tuberculosis strains. PMID- 15364988 TI - Risk factors for community-associated methicillin-resistant Staphylococcus aureus infections in an outbreak of disease among military trainees in San Diego, California, in 2002. AB - An outbreak of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) skin infections was observed in a population of U.S. military trainees in the summer of 2002. A questionnaire was developed and administered to 206 trainees, 22 of whom had MRSA infections. Factors associated with infection were described by multivariable logistic regression modeling and included having a roommate in training with a prior skin infection (odds ratio [OR] = 3.44) or having a family member or friend who worked in a health care setting (OR = 2.79). Previous antibiotic use, hospitalization, or health problems were not associated with MRSA infection. This outbreak of MRSA skin infections in an otherwise healthy, well-defined, military population provided an opportunity to describe risk factors for CA-MRSA which may help focus prevention efforts in this and other communities. PMID- 15364989 TI - Evaluation of the core antigen assay as a second-line supplemental test for diagnosis of active hepatitis C virus infection. AB - The British Columbia Center for Disease Control laboratory performs approximately 95% of all hepatitis C virus (HCV) antibody tests for the province's 4 million inhabitants. In 2002, the laboratory tested 96,000 specimens for anti-HCV antibodies, of which 4,800 (5%) were seroreactive and required confirmation of active infection. Although HCV RNA assays with a sensitivity of 50 IU/ml or less are recommended for the confirmation of active HCV infection, given the large number of seroreactive specimens tested annually, we evaluated the Ortho trak-C assay (OTCA) as a second-line confirmatory test and determined its limit of detection (LoD). Of 502 specimens from treatment-naive anti-HCV-positive individuals, 478 had sufficient volumes for evaluation by the OTCA and HCV RNA tests. Core antigen was not detected in 147 of 478 (30.8%) of these specimens, of which 37 of 147 (25.2%) were shown to be viremic by the VERSANT HCV (version 3.0) (branched-DNA) assay and/or the VERSANT HCV qualitative assay. Testing of 144 replicates of a World Health Organization standard dilution series indicated that the LoD of OTCA was approximately 27,000 IU/ml. This LoD is consistent with the inability of OTCA to detect core antigen in clinical specimens with low viral loads. We conclude that OTCA has limited value as a confirmatory test for the diagnosis of active HCV infection because 37 of 367 (10%) of viremic specimens had undetectable core antigen. Qualitative HCV RNA testing remains the present standard for the confirmation of active HCV infection in the diagnostic setting. PMID- 15364990 TI - Evaluation of Trichinella spiralis larva group 1 antigens for serodiagnosis of human trichinellosis. AB - To identify Trichinella antigens suitable for high-specificity and high sensitivity serodiagnosis of human trichinellosis, we evaluated assays using four antigens: (i) crude first-stage larval extract (CLE), (ii) O-deglycosylated CLE, (iii) tyvelose-bearing antigens (Trichinella spiralis larva group 1 [TSL-1] antigens) purified by US4 affinity chromatography and coupled directly to enzyme linked immunosorbent assay (ELISA) plates (pTSL-1 antigens), and (iv) TSL-1 antigens immobilized on ELISA plates with the monoclonal antibody (MAb) US4 (cTSL 1 antigens). Assays using these antigens were compared by analysis of sera from healthy individuals (n = 224) (group 1), individuals with noninfectious intestinal pathologies (n = 114) (group 2), individuals with other parasitic infections (n = 107) (group 3), and individuals with confirmed trichinellosis (n = 42) (group 4). Our results indicate that capture ELISA using cTSL-1 antigens is the most effective method for serodiagnosis of human trichinellosis; this was the only method showing 100% specificity and 100% sensitivity at the patent stage of the infection, and it was also the most sensitive for sera obtained prior to patency in indirect immunofluorescence (IIF). Indirect ELISA with pTSL-1 antigens was also 100% specific but was slightly less sensitive, particularly with sera obtained before IIF patency. Inhibition ELISA with MAb US4 indicated (i) that in Trichinella-infected patients the immune response to TSL-1 antigens is directed mostly against tyvelose-containing epitopes (mean of 84.2% of total anti-TSL-1 immunoglobulin G1 [IgG1] antibody response [range, 51.3 to 97.6%]) and (ii) that in most individuals a large proportion of anti-CLE IgG1 antibodies (mean, 49.5%; range, 7.3 to 92.6%) are directed against tyvelose epitopes. PMID- 15364991 TI - Evaluation of the new VITEK 2 card for identification of clinically relevant gram negative rods. AB - The VITEK 2 card for gram-negative bacteria (bioMerieux,Marcy-l'Etoile, France) has been redesigned to improve the identification of fermenting and nonfermenting bacilli. Forty-seven biochemical tests, including 19 enzymatic tests, are present in the new card and interpreted in a kinetic mode. Final identification results are available within 10 h. The database allows the identification of 159 different taxa. Six hundred fifty-five gram-negative rods (GNR; 511 fermenters and 144 nonfermenters), representing 54 taxa, were tested. Strains were taken from fresh routine primary isolation plates (n = 157), from stored routine plates (n = 301), and from stock cultures (n = 197). Six hundred thirty-seven strains (97.3%) were correctly identified to the species level, 14 strains (2.1%) gave low discrimination results requiring additional tests, and 4 strains (0.6%) gave discordant results; not a single strain remained unidentified. Nearly 92% of all isolates were correctly identified within 7 h of incubation. The robustness of the system was demonstrated by the fact that strains were grown on four different agar media before testing. The system may also have the potential to be applied directly to primary isolation plates, since in this instance 96.2% of 157 GNR were correctly identified and 3.8% gave low discrimination results. The new VITEK 2 card for gram-negative bacteria seems to be a promising new tool for routine, rapid identification of GNR. PMID- 15364992 TI - Rapid detection and identification of human adenovirus species by adenoplex, a multiplex PCR-enzyme hybridization assay. AB - Human adenoviruses (AdV) have been implicated in a wide variety of diseases and are ubiquitous in populations worldwide. These agents are of concern particularly in immunocompromised patients, children, and military recruits, resulting in severe disease or death. Clinical diagnosis of AdV is usually achieved through routine viral cell culture, which can take weeks for results. Immunofluorescence and enzyme-linked immunosorbent assay-based techniques are more timely but lack sensitivity. The ability to distinguish between the six different AdV species (A to F) is diagnostically relevant, as infections with specific AdV species are often associated with unique clinical outcomes and epidemiological features. Therefore, we developed a multiplex PCR-enzyme hybridization assay, the Adenoplex, using primers to the fiber gene that can simultaneously detect all six AdV species A through F in a single test. The limit of detection (LOD) based on the viral 50% tissue culture infective dose/ml for AdV A, B, C, D, E, and F was 10(-2), 10(-1), 10(-1), 10(-2), 10(-1), and 10(-2), respectively. Similarly, the LOD for the six DNA controls ranged from 10(2) to 10(3) copies/ml. Twelve common respiratory pathogens were tested with the Adenoplex, and no cross-reactivity was observed. We also validated our assay using clinical specimens spiked with different concentrations of AdV strains of each species type and tested by multiplex PCR and culture. The results demonstrated an overall sensitivity and specificity of Adenoplex of 100%. This assay can be completed in as few as 5 h and provides a rapid, specific, and sensitive method to detect and subtype AdV species A through F. PMID- 15364993 TI - Isoenzymatic analysis of 712 strains of Leishmania infantum in the south of France and relationship of enzymatic polymorphism to clinical and epidemiological features. AB - In the south of France, leishmaniasis due to Leishmania infantum occurs in the following five foci of endemicity (from west to east): Pyrenees-Orientales, Cevennes, Provence, Cote d'Azur, and Corsica. Between 1981 and 2002, 712 Leishmania strains obtained from humans, dogs, cats, and sand flies were studied by isoenzyme analysis. In total, seven zymodemes were identified: MON-1, MON-11, MON-24, MON-29, MON-33, MON-34, and MON-108. The Pyrenees-Orientales focus is characterized by a predominance of human cutaneous leishmaniasis and a high enzymatic polymorphism (five zymodemes). In the other foci, where human visceral leishmaniasis is predominant, only two zymodemes are present. L. infantum MON-1 is the parasite most frequently found, in patients both with and without concomitant human immunodeficiency virus infection. MON-1 is the only zymodeme present in dogs, which act as the reservoir host in all of the foci. In Cevennes, where the complete life cycle of zymodeme MON-1 has been identified, Phlebotomus perniciosus and Phlebotomus ariasi are vectors. The enzymatic polymorphism is compared to that of neighboring countries (Spain and Italy). In Pyrenees Orientales, small variant zymodemes with electromorphs of heterozygote-like and homozygotic patterns can be explained by different genetic hypotheses. PMID- 15364994 TI - Development and validation of a diagnostic DNA microarray to detect quinolone resistant Escherichia coli among clinical isolates. AB - The incidence of resistance against fluoroquinolones among pathogenic bacteria has been increasing in accordance with the worldwide use of this drug. Escherichia coli is one of the most relevant species for quinolone resistance. In this study, a diagnostic microarray for single-base-mutation detection was developed, which can readily identify the most prevalent E. coli genotypes leading to quinolone resistance. Based on genomic sequence analysis using public databases and our own DNA sequencing results, two amino acid positions (83 and 87) on the A subunit of the DNA gyrase, encoded by the gyrA gene, have been identified as mutation hot spots and were selected for DNA microarray detection. Oligonucleotide probes directed against these two positions were designed so that they could cover the most important resistance-causing and silent mutations. The performance of the array was validated with 30 clinical isolates of E. coli from four different hospitals in Germany. The microarray results were confirmed by standard DNA sequencing and were in full agreement with phenotypic antimicrobial susceptibility testing. PMID- 15364995 TI - Real-time fluorescence PCR assays for detection and characterization of heat labile I and heat-stable I enterotoxin genes from enterotoxigenic Escherichia coli. AB - To facilitate the diagnosis of enterotoxigenic Escherichia coli (ETEC) infections in humans, we developed and evaluated real-time fluorescence PCR assays for the Roche LightCycler (LC) against the enterotoxin genes commonly present in strains associated with human illness. Separate LC-PCR assays with identical cycling conditions were designed for the type I heat-labile enterotoxin (LT I) and the type I heat-stable enterotoxin (ST I) genes, using the LC hybridization probe format. A duplex assay for ST I with two sets of amplification primers and three hybridization probes was required to detect the major nucleotide sequence variants of ST I, ST Ia and ST Ib. LC-PCR findings from the testing of 161 E. coli isolates of human origin (138 ETEC and 23 non-ETEC) were compared with those obtained by block cycler PCR analysis. The sensitivities and specificities of the LC-PCR assays were each 100% for the LT I and ST I genes. The LC-PCR and block cycler PCR assays were also compared for their abilities to detect LT I and ST I genes in spiked stool specimens with different methods of sample preparation. Findings from these experiments revealed that the limits of detection for the LC PCR assays were the same or substantially lower than those observed for the block cycler PCR assay. Melting curve analysis of the amplified LT I and ST I genes revealed sequence variation within each gene, which for the ST I genes correlated with the presence of ST Ia and ST Ib. The rapidity, sensitivity, and specificity of the LC-PCR assays make them attractive alternatives to block cycler PCR assays for the detection and characterization of ETEC. PMID- 15364996 TI - Use of recombinant E protein domain III-based enzyme-linked immunosorbent assays for differentiation of tick-borne encephalitis serocomplex flaviviruses from mosquito-borne flaviviruses. AB - The serological diagnosis of infection by flaviviruses is complicated by the presence of flavivirus cross-reactive antibodies that produce false-positive results for flavivirus infections, especially in regions where more than one virus is endemic. Current diagnostic reagents for tick-borne flavivirus infection have been found to cross-react with yellow fever- or dengue virus-positive sera. This study utilized recombinant flavivirus E protein domain 3 (rE-D3) as a diagnostic reagent to differentiate between infection by mosquito- and tick-borne flaviviruses. This study found that the use of rE-D3 in an enzyme-linked immunosorbent assay (ELISA)-based format allowed the differentiation between serum specific for either mosquito- or tick-borne flaviviruses, but not among the members of the tick-borne encephalitis (TBE) serocomplex of flaviviruses. Sera derived against several TBE serocomplex rE-D3 were found to cross-react with heterologous rE-D3 within the TBE serocomplex, but not with those from mosquito borne flaviviruses, in both Western blots and ELISAs. Mouse hyperimmune sera generated against TBE serocomplex viruses were also found to react specifically with TBE serocomplex rE-D3, but not with rE-D3 from mosquito-borne viruses and vice versa. When a similar test using virus-derived antigen was performed, a loss of both specificity and sensitivity was observed. These results indicate that flavivirus rE-D3 would be a useful reagent for the detection of infection by TBE serocomplex flaviviruses, several of which are potential biothreat agents, but would not provide the ability to differentiate among infections by separate members of the serocomplex. PMID- 15364997 TI - Virulence characteristics and molecular epidemiology of enteroaggregative Escherichia coli isolates from hospitalized diarrheal patients in Kolkata, India. AB - Enteroaggregative Escherichia coli (EAEC) is an important diarrheal enteropathogen defined by aggregative adherence to cultured epithelial cells. We have detected EAEC from 121 (6.6%) of 1,826 hospitalized patients admitted with diarrhea to the Infectious Diseases Hospital in Kolkata, India. Watery diarrhea was recorded significantly (P = 0.0142) more often in children. The majority of the EAEC isolates were not serotypeable (62%) and showed resistance to five or more antibiotics (76%). We studied different virulence genes and the molecular epidemiology of 121 EAEC isolates recovered from diarrheal patients. A PCR assay for detection of virulence genes, an assay for determination of clump formation in liquid culture, and a HeLa cell adherence assay were carried out to characterize the EAEC isolates. Investigations were also conducted to correlate the virulence gene profiles with diarrheal symptoms and molecular epidemiology by pulsed-field gel electrophoresis (PFGE). Two or more virulence genes were detected in 109 (90.1%) EAEC isolates. In the cluster analysis, some isolates with specific gene profiles and phenotypes formed a group or subcluster. This study highlights the comparative distributions of three fimbrial adhesins and other virulence genes among EAEC isolates. The diverse virulence gene and PFGE profiles, along with the existence of diverse serotypes and antibiograms, suggests that the EAEC isolates are genetically heterogeneous in Kolkata. PMID- 15364998 TI - Correlation of wbiI genotype, serotype, and isolate source within species of the Burkholderia cepacia complex. AB - Gram-negative bacteria of the Burkholderia cepacia complex (Bcc) are opportunistic pathogens that can infect the lungs of cystic fibrosis (CF) patients and can be transmitted among these patients, causing epidemics in the CF community. Lipopolysaccharide (LPS) is an important virulence factor of many gram negative bacteria, with the O antigen component of LPS being responsible for serotype specificity. The goal of this work was to develop a genetic method of determining the serotype of Bcc isolates based on the conserved gene wbiI. Homologues of wbiI are found in polysaccharide biosynthesis gene clusters in other bacteria. Primers to a conserved region of the Bcc wbiI gene were able to amplify by PCR a single product in 67 of 80 Bcc isolates tested. Sequencing and restriction enzyme digestion of this wbiI PCR product revealed sufficient DNA polymorphisms to distinguish and group various isolates. In five of nine instances, Bcc isolates of a single serotype had a single wbiI restriction fragment length polymorphism (RFLP) pattern, while isolates of the other four serotypes could have multiple wbiI RFLP types. Species determination of the Bcc isolates revealed no obvious correlation between wbiI RFLP type and species. There was also no apparent correlation between wbiI RFLP type and the ability of a single Bcc isolate to infect an individual with CF. However three of five Bcc outbreaks involved isolates with the same wbiI RFLP type, indicating that wbiI RFLP typing may be a useful tool to help track Bcc outbreaks. PMID- 15364999 TI - Isolation of metronidazole-resistant Bacteroides fragilis carrying the nimA nitroreductase gene from a patient in Washington State. AB - Members of the Bacteroides fragilis group are among the most common anaerobic bacterial isolates in clinical specimens. Metronidazole, a 5-nitroimidazole, is often used as empirical therapy for anaerobic infections. Susceptibility testing is not routinely performed because of nearly universal susceptibility of Bacteroides spp. to this agent. We report a case of metronidazole-resistant Bacteroides fragilis in the United States and demonstrate the presence of the nimA gene, encoding a nitroreductase previously shown to mediate resistance to 5 nitroimidazole antimicrobial agents in B. fragilis strains from Europe and Africa. Because clinical failures in Bacteroides infections have been associated with the use of inactive antimicrobial agents, clinicians need to be aware of the possibility of metronidazole-resistant B. fragilis strains in the United States and the importance of susceptibility testing in selected situations. PMID- 15365000 TI - Use of the MagNA pure LC automated nucleic acid extraction system followed by real-time reverse transcription-PCR for ultrasensitive quantitation of hepatitis C virus RNA. AB - Hepatitis C virus (HCV) infection is an increasing health problem worldwide. Quantitative assays for HCV viral load are valuable in predicting response to therapy and for following treatment efficacy. Unfortunately, most quantitative tests for HCV RNA are limited by poor sensitivity. We have developed a convenient, highly sensitive real-time reverse transcription-PCR assay for HCV RNA. The assay amplifies a portion of the 5' untranslated region of HCV, which is then quantitated using the TaqMan 7700 detection system. Extraction of viral RNA for our assay is fully automated with the MagNA Pure LC extraction system (Roche). Our assay has a 100% detection rate for samples containing 50 IU of HCV RNA/ml and is linear up to viral loads of at least 10(9) IU/ml. The assay detects genotypes 1a, 2a, and 3a with equal efficiency. Quantitative results by our assay correlate well with HCV viral load as determined by the Bayer VERSANT HCV RNA 3.0 bDNA assay. In clinical use, our assay is highly reproducible, with high and low control specimens showing a coefficient of variation for the logarithmic result of 2.8 and 7.0%, respectively. The combination of reproducibility, extreme sensitivity, and ease of performance makes this assay an attractive option for routine HCV viral load testing. PMID- 15365001 TI - Identification of the emerging pathogen Vibrio vulnificus biotype 3 by commercially available phenotypic methods. AB - Identification of the emerging pathogen Vibrio vulnificus biotype 3 has become a challenge for clinical laboratories in the last few years. In this study, the abilities of five commercial systems to identify this new species have been evaluated for the first time, using a unique collection of strains. Fifty-one well-documented wild strains of V. vulnificus biotype 3 were processed using API 20 NE, GNI+ Vitek 1 cards, ID-GNB Vitek 2 cards, Neg Combo 20 Microscan panels, and NMIC/ID-5 BD Phoenix panels. The numbers of strains identified as V. vulnificus by ID-GNB, NMIC/ID-5, and GNI+ were 50 (98.0%), 46 (90.2%), and 7 (13.7%), respectively. Neg Combo 20 Microscan panels and API 20 NE were unable to identify any of the strains of this emerging pathogen to the species level and mostly misidentifies them as other species of the Vibrionaceae family. Data on the phenotypic pattern of V. vulnificus biotype 3 when processed in all five systems as presented here could help clinical laboratories in identifying this new pathogen. PMID- 15365002 TI - Prevalence of periodontal pathogens in dental plaque of children. AB - Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Tannerella forsythensis have been implicated as the main etiological agents of periodontal disease. The purpose of this work was to estimate the prevalence of these organisms in plaque from children without gingivitis (group 1; n = 65) and from those with gingivitis (group 2; n = 53). Extracted DNA from plaque was subjected to two rounds of PCR targeting the 16S rRNA gene using both universal primers and species-specific primers. The results were as follows: group 1, P. gingivalis, 49%; A. actinomycetemcomitans, 55%; and T. forsythensis, 65%; group 2, P. gingivalis, 47%; A. actinomycetemcomitans, 59%; and T. forsythensis, 45%. T. forsythensis was detected more frequently in children with no gingivitis than in those with gingivitis (P = 0.03). There was no significant difference between the two groups with respect to the presence of P. gingivalis or A. actinomycetemcomitans in either group (P > 0.05). Logistic regression analysis revealed that the odds of a patient having gingivitis were 2.3 times greater in the absence of T. forsythensis. In conclusion, the results of this study have shown that the three pathogens can be detected in the dental plaque of healthy children and of those with gingivitis and that T. forsythensis is associated with dental plaque at sites with no gingivitis. PMID- 15365003 TI - Field evaluation of a rapid human immunodeficiency virus (HIV) serial serologic testing algorithm for diagnosis and differentiation of HIV type 1 (HIV-1), HIV-2, and dual HIV-1-HIV-2 infections in West African pregnant women. AB - We evaluated a two-rapid-test serial algorithm using the Determine and Genie II rapid assays, performed on-site in four peripheral laboratories during the French Agence Nationale de Recherches sur le SIDA (ANRS) 1201/1202 Ditrame Plus cohort developed for prevention of mother-to-child transmission of human immunodeficiency virus (HIV) infection in Cote d'Ivoire. A total of 1,039 specimens were retested by two commercial enzyme-linked immunosorbent assays (ELISAs). The following specimens were tested: 315 specimens found on-site to be infected with HIV type 1 (HIV-1), 8 specimens found on-site to be infected with HIV-2, 71 specimens found on-site to be infected with both HIV-1 and HIV-2, 40 specimens found on-site to have indeterminate results for HIV infection, and 605 specimens taken during a quality assurance program. For HIV discrimination, 99 positive serum samples (20 with HIV-1, 8 with HIV-2, and 71 with HIV-1 and HIV-2 on the basis of our rapid test algorithm) were retested by the Peptilav test, Western blot (WB) assays, and homemade monospecific ELISAs. Real-time DNA PCRs for the detection of HIV-1 and HIV-2 were performed with peripheral blood mononuclear cells from 35 women diagnosed on-site with HIV-1 and HIV-2 infections. Compared to the results of the ELISAs, the sensitivities of the Determine and Genie II assays were 100% (95% lower limit [95% LL], 99.1%) and 99.5% (95% confidence interval [95% CI], 98.2 to 99.9%), respectively. The specificities were 98.4% (95% CI, 96.9 to 99.3%) and 100% (95% LL, 99.3%), respectively. All serological assays gave concordant results for infections with single types. By contrast, for samples found to be infected with dual HIV types by the Genie II assay, dual reactivity was detected for only 37 samples (52.1%) by WB assays, 34 samples (47.9%) by the Peptilav assay, and 23 samples (32.4%) by the monospecific ELISAs. For specimens with dual reactivity by the Genie II assay, the rates of concordance between the real-time PCR assays and the serological assays were 25.7% for the Genie II assay, 82.9% for the Peptilav assay, 74.3% for WB assays, and 80% for the homemade ELISAs. Our algorithm provided high degrees of sensitivity and specificity comparable to those of ELISAs. Even if they are rare, women identified by the Genie II assay as being infected with HIV-1 and HIV-2 mostly appeared to be infected only with HIV-2. PMID- 15365004 TI - Diagnosing invasive aspergillosis during antifungal therapy by PCR analysis of blood samples. AB - We evaluated the value of Aspergillus PCR as a tool for diagnosing invasive aspergillosis from whole-blood samples during antifungal therapy. In a 3-year study, 36 patients receiving antifungal therapy due to chest radiographic findings highly suggestive of fungal pneumonia were evaluated. The PCR results from whole-blood samples were compared to those obtained from bronchoalveolar lavage fluids and/or tissue specimens. A total of 205 whole-blood samples, 15 fine-needle aspirations or tissue biopsy specimens, and 21 bronchoalveolar lavage fluids and tracheal secretions were analyzed using PCR. Of the 36 patients, 15 had proven, 9 had probable, and 12 had possible invasive Aspergillus infection according to European Organization for Research and Treatment of Cancer/Mycosis Study Group definitions. For patients with proven infection the sensitivity values of PCR in lung and blood samples were 100 and 40%, respectively. The negative predictive value of blood monitoring under conditions of antifungal treatment was 44%. Clearance of fungal DNA from blood was associated with resolution of clinical symptoms in six of nine patients with proven infection. Repeated positive PCR results for Aspergillus were associated with fatal outcome, as three of six patients died. For patients with probable infection the sensitivity values of PCR in lung fluid and blood were 66 and 44%, respectively. The benefit of PCR diagnosis using whole-blood samples is limited when sampling takes place after treatment has been started. Performance of Aspergillus PCR using tissue samples is recommended in addition to microscopic examination and culture technique for sensitive detection of fungal infection. PMID- 15365005 TI - Continuing diversification of Neisseria meningitidis W135 as a primary cause of meningococcal disease after emergence of the serogroup in 2000. AB - The occurrence of a clonal outbreak of serogroup W135 (of the electrophoretic type 37 [ET-37] clonal complex) meningococcal disease among Hajj pilgrims in 2000 has led to enhanced surveillance of the evolution of this particular serogroup, formerly considered rare, in invasive infections. Since the first case of meningococcal disease due to a serogroup W135 strain was detected in France in 1994, all isolates were characterized phenotypically. We further used phenotypic and genotypic approaches to type the 101 serogroup W135 strains isolated from patients with invasive meningococcal diseases in France in 2001 and 2002. Overall, 55% of these isolates had Hajj strain-related phenotypes (60 and 52% in 2001 and 2002, respectively), although only 45% belonged to the ET-37 clonal complex. Moreover, pulsed-field gel electrophoresis of the ET-37 clonal complex isolates showed that only 32% of the serogroup W135 isolates were indistinguishable from the 2000 Hajj-related strain. Our results suggest the continuous emergence of new genetic lineages of serogroup W135 independently of the 2000 global outbreak. PMID- 15365006 TI - A predominant and virulent Legionella pneumophila serogroup 1 strain detected in isolates from patients and water in Queensland, Australia, by an amplified fragment length polymorphism protocol and virulence gene-based PCR assays. AB - In epidemiological investigations of community legionellosis outbreaks, knowledge of the prevalence, distribution, and clinical significance (virulence) of environmental Legionella isolates is crucial for interpretation of the molecular subtyping results. To obtain such information for Legionella pneumophila serogroup 1 isolates, we used the standardized amplified fragment length polymorphism (AFLP) protocol of the European Working Group on Legionella Infection to subtype L. pneumophila SG1 isolates obtained from patients and water sources in Queensland, Australia. An AFLP genotype, termed AF1, was predominant in isolates from both patients (40.5%) and water (49.0%). The second most common AFLP genotype found in water isolates was AF16 (36.5%), but this genotype was not identified in the patient isolates. When virulence gene-based PCR assays for lvh and rtxA genes were applied to the isolates from patients and water, nearly all (65 of 66) AF1 strains had both virulence genes, lvh and rtxA. In contrast, neither the lvh nor the rtxA gene was found in the AF16 strains, except for one isolate with the rtxA gene. It appears that this may explain the failure to find this genotype in the isolates from patients even though it may be common in the environment. In view of the evidence that the AF1 genotype is the most common genotype among strains found in patients and water sources in this region, any suggested epidemiological link derived from comparing the AF1 genotype from patient isolates with the AF1 genotype from environmental isolates must be interpreted and acted on with caution. The use of virulence gene-based PCR assays applied to environmental samples may be helpful in determining the infection potential of the isolates involved. PMID- 15365007 TI - Complementation in cells cotransfected with a mixture of wild-type and mutant human immunodeficiency virus (HIV) influences the replication capacities and phenotypes of mutant variants in a single-cycle HIV resistance assay. AB - The impact of cotransfection of mixtures of mutant and wild type (WT) virus on the observed phenotype and replication capacity (RC) in a single-cycle human immunodeficiency virus (HIV) phenotypic assay has been investigated by cotransfecting mutant HIV clones expressing the firefly luciferase expression gene with a WT clone expressing Renilla luciferase. Four mutant constructs with different genotypes displayed <1% RC when transfected alone. Cotransfection of as little as 9% of the WT clone resulted in an 18- to 33-fold increase in the RC of the mutant clones. In addition, the 50% inhibitory concentration (IC(50)) of lopinavir against seven mutant clones decreased by up to 97% after incremental cotransfection of 9 to 50% of the WT clone. The enhancement of RC and decrease in IC(50) for mutant variants following cotransfection with the WT variant appear to be due to complementation rather than genetic recombination. These findings suggest that the RC and susceptibility of plasma isolates from patients who are off therapy or not adherent to treatment, in which WT virus may expand to significant levels, should be interpreted with caution. PMID- 15365008 TI - Automated high-throughput mycobacterial interspersed repetitive unit typing of Mycobacterium tuberculosis strains by a combination of PCR and nondenaturing high performance liquid chromatography. AB - Mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU VNTR) typing of Mycobacterium tuberculosis complex isolates is portable, 100% reproducible, and highly discriminatory. Nondenaturing high-performance liquid chromatography (non-dHPLC) with use of a WAVE microbial analysis system is a promising method of PCR amplicon analysis as it is low cost and requires no preanalysis processing. The aims of this study were to validate the application of WAVE microbial analysis system technology to MIRU-VNTR typing. A collection of 70 strains were cultivated in liquid culture and extracted using the QIAamp DNA minikit. Novel primers were designed to target the 12 MIRU-VNTR loci (P. Supply et al., J. Clin. Microbiol. 39:3563-3571, 2001). After amplification, each PCR product was analyzed on a WAVE microbial analysis system. The fragment size was calculated from the chromatogram, and the number of tandem repeats at each locus was determined. For the collection of 70 strains 100% concordance was achieved when comparing MIRU-VNTR profiles obtained from agarose gel electrophoresis and PCRs analyzed on a WAVE microbial analysis system. The calculated fragment sizes, obtained from the WAVE microbial analysis system, were sufficiently accurate to ensure 100% confidence when assigning the number of tandem repeats to a MIRU-VNTR locus. This study is the first to report the successful use of non-dHPLC for screening for variations in the number of MIRU-VNTRs in mycobacterial DNA. Non dHPLC analysis was demonstrated to be a rapid, low-labor input method for the detection and analysis of MIRU-VNTR amplicons. The combination with non-dHPLC further enhances the utility of MIRU-VNTR typing. PMID- 15365009 TI - Oligonucleotide chip for detection of Lamivudine-resistant hepatitis B virus. AB - Hepatitis B virus (HBV) is one of the major causes of liver disease worldwide. It is important to conduct antiviral therapy against chronic hepatitis B to minimize the amount of liver damage. Lamivudine has been known to be an effective antiviral agent for the treatment of HBV infection. However, the emergence of viral mutants resistant to lamivudine is the main concern during the treatment of HBV-infected patients. Therefore, the detection of lamivudine-resistant mutants is of clinical importance. We have developed an oligonucleotide chip for the detection of lamivudine-resistant HBV which is rapid and accurate. The oligonucleotide chip consists of quality control probes, negative control probes, and specific oligonucleotide probes for the detection of lamivudine-resistant HBV. The specific probes consist of five probes for the detection of wild-type rtL180, rtM204, and rtV207 sequences and seven probes for the detection of HBV mutations. We tested 123 serum samples from patients with chronic HBV infection who had received lamivudine therapy. Eighty samples contained mutants with YMDD mutations. Among these, 17 contained rtM204V (YVDD), 24 contained rtM204I3 (YIDD3), 3 contained rtM204I2 (YIDD2), and 36 contained mixed types. We compared the results obtained with our oligonucleotide chip with those obtained by PCR restriction fragment length polymorphism (RFLP) analysis and sequencing. The rate of concordance between the assay with the oligonucleotide chip and PCR-RFLP analysis for detection of the YMDD motif was 96.7%. The rate of concordance between the results obtained with the oligonucleotide chip for the detection of rtL180 and rtV207 and the results obtained by sequencing was 100%. Thus, the oligonucleotide chip is a reliable and useful tool for the detection of antiviral resistant HBV. PMID- 15365010 TI - Identification of some charcoal-black-pigmented CDC fermentative coryneform group 4 isolates as Rothia dentocariosa and some as Corynebacterium aurimucosum: proposal of Rothia dentocariosa emend. Georg and Brown 1967, Corynebacterium aurimucosum emend. Yassin et al. 2002, and Corynebacterium nigricans Shukla et al. 2003 pro synon. Corynebacterium aurimucosum. AB - Sixty-three clinical isolates of charcoal-black-pigmented, gram-positive coryneform rods were received for identification by the Centers for Disease Control and Prevention (CDC) and were provisionally designated CDC fermentative coryneform group 4 (FCG4). Forty-five of these were characterized by morphological, physiologic, antimicrobial susceptibility, cellular fatty acids, 16S rRNA gene sequencing, and DNA-DNA hybridization analyses. Nitrate reduction, cellular fatty acid analysis, 16S rRNA gene sequencing, and DNA-DNA hybridization studies segregated these strains into two groups: FCG4a (8 strains) and FCG4b (37 strains). The FCG4a strains, only one of which was from a female genitourinary source, produced cellular fatty acid and biochemical profiles similar to those observed with reference strains of Rothia dentocariosa and Rothia mucilaginosa, while the FCG4b strains were similar to Corynebacterium species. DNA-DNA hybridization analysis demonstrated species-level relatedness among six FCG4a tested strains and showed that they were a charcoal-black-pigmented variant of R. dentocariosa. Sixteen isolates of the FCG4b group, mainly from female genitourinary tract specimens, as well as the type strains of two recently named species, Corynebacterium aurimucosum and Corynebacterium nigricans, were shown by DNA-DNA hybridization analysis and the sequencing of the 16S rRNA gene to be related at the species level and unrelated to the type strain of R. dentocariosa; therefore, the Corynebacterium-like strains were classified as a charcoal-black pigmented variant of C. aurimucosum, because this name has nomenclatural priority over C. nigricans. These findings indicate that FCG4 represents a heterogeneous group that contains pigmented variants of both R. dentocariosa and C. aurimucosum; hence, the descriptions of both R. dentocariosa and C. aurimucosum have been amended to include charcoal-black-pigmented variants, and C. nigricans is a pro synonym of C. aurimucosum. PMID- 15365011 TI - Correlation of cerebrospinal fluid (CSF) cell counts and elevated CSF protein levels with enterovirus reverse transcription-PCR results in pediatric and adult patients. AB - During the 2001, 2002, and 2003 enterovirus seasons, we investigated the correlations between cerebrospinal fluid (CSF) nucleated cell counts and elevated CSF protein levels and the detection of enteroviral RNA by reverse transcription (RT)-PCR. Our objective was to determine if pleocytosis and/or elevated protein levels were predictive of positive RT-PCR results for enterovirus. We were also interested in determining if the presence of West Nile virus during the 2002 enteroviral season contributed to a change in these correlations. We found that in the group of patients aged >2 months, the absence of pleocytosis was highly predictive of a negative RT-PCR result. Elevated CSF protein level was not a good predictor of RT-PCR positivity for enterovirus and did not add to the diagnostic sensitivity or specificity of pleocytosis. PMID- 15365012 TI - Rapid detection of isoniazid and rifampin resistance mutations in Mycobacterium tuberculosis complex from cultures or smear-positive sputa by use of molecular beacons. AB - The slow-growing nature of Mycobacterium tuberculosis complex hinders the improvement of turnaround time for phenotypic drug susceptibility testing. We designed a set of molecular beacons for the detection of isoniazid and rifampin resistance mutations in M. tuberculosis complex organisms from cultures or from N acetyl-l-cysteine-NaOH-treated, smear-positive specimens. The performance of the molecular beacons was characterized by studying a total of 196 clinical isolates (127 drug-resistant isolates and 69 drug-susceptible isolates). For detection of isoniazid resistance, the sensitivity and specificity of the assay were 82.7 and 100%, and the positive predictive value (PPV) and negative predictive value (NPV) at a resistance prevalence of 10% were 100 and 98.11%, respectively. For detection of rifampin resistance, the sensitivity and specificity of the assay were 97.5 and 100%, and the PPV and NPV at a resistance prevalence of 2.0% were 100 and 99.95%, respectively. PMID- 15365013 TI - Laboratory reporting of tuberculosis test results and patient treatment initiation in California. AB - Prompt laboratory reporting of tuberculosis (TB) test results is necessary for TB control. To understand the extent of and factors contributing to laboratory reporting delays and the impact of reporting delays on initiation of treatment of TB patients, we analyzed data from 300 consecutive culture-positive TB cases reported in four California counties in 1998. Laboratory reporting to the specimen submitter was delayed for 26.9% of smear-positive patients and 46.8% of smear-negative patients. Delays were associated with the type of laboratory that performed the testing and with delayed transport of specimens. Referral laboratories (public health and commercial) had longer median reporting time frames than hospital and health maintenance organization laboratories. Among patients whose treatment was not started until specimens were collected, those with delayed laboratory reporting were more likely to have delayed treatment than patients with no laboratory reporting delays (odds ratio [OR] of 3.9 and 95% confidence interval [CI] of 1.6 to 9.7 for smear-positive patients and OR of 13.1 and CI of 5.3 to 32.2 for smear-negative patients). This relation remained after adjustment in a multivariate model for other factors associated with treatment delays (adjusted OR of 25.64 and CI of 7.81 to 83.33 for smear-negative patients). These findings emphasize the need to reduce times of specimen transfer between institutions and to ensure rapid communication among laboratories, health care providers, and health departments serving TB patients. PMID- 15365014 TI - Phenotypic and molecular characteristics of Streptococcus agalactiae isolates recovered from milk of dairy cows in Brazil. AB - Information on the characteristics of Streptococcus agalactiae obtained from bovine sources in Brazil is still very limited. The aim of this study was to assess the phenotypic and genotypic diversity among S. agalactiae isolates from milk of dairy cows presenting clinical or subclinical mastitis in the southeast region of Brazil. Phenotypic characterization was based on physiological and serological tests. Antimicrobial susceptibility tests were carried out by the disk method. Genetic diversity was evaluated by using random amplified polymorphic DNA-PCR (RAPD-PCR) (by using the primer 1254) and pulsed-field gel electrophoresis (PFGE) (by using SmaI as the restriction enzyme) and by PCRs for detection of genes associated with resistance to erythromycin and tetracycline as well as PCRs for detection of genes coding for cell surface-associated proteins. According to the results of physiologic tests, 45 (52.9%) isolates showed beta hemolysis and 44 (51.7%) were susceptible to bacitracin. Fourteen different biotypes were detected. The two most frequent biotypes comprised strains that were non-beta-hemolytic; fermented galactose, lactose, and salicin; produced protease; and were negative for DNase production. Serotype III was predominant (66 isolates [77.6%]), followed by serotypes II, Ia, Ib, and VI. Resistance to tetracycline and erythromycin was found in 38 (44.7%) and 9 (10.5%) isolates, respectively, with tet(O) (31.7%) and erm(B) (100%) being the most frequently occurring resistance genes. Three genes coding for surface proteins, bca, lmb, and scpB, were detected in 55 (64.7%), 7 (8.2%), and 43 (50.5%) isolates, respectively. In most cases, isolates from animals in the same herd presented closely related genetic profiles (determined by either RAPD-PCR or PFGE), which were distinct from those of isolates from different herds. PMID- 15365015 TI - Accurate representation of the hepatitis C virus quasispecies in 5.2-kilobase amplicons. AB - Hepatitis C virus (HCV) exists as a swarm of genetically distinct but related variants, or a quasispecies, whose complexity and sequence evolution are critical to studies of viral pathogenesis. Because most studies of the HCV quasispecies have focused on a relatively small genomic segment, the first hypervariable region of the E2 gene, it is possible that viral complexity is occasionally underestimated (due to primer mismatch) and that sequence evolution is misperceived due to unrecognized covariation. This report describes a sensitive and reproducible method to amplify most of the HCV genome as a single 5.2-kb amplicon by using primers directed at relatively conserved genomic segments. Using 52 specimens obtained during acute infection over a range of viral RNA concentrations, the overall rate of successful amplification was 94% and varied in a concentration-dependent manner, with successful amplification in 26 of 26 (100%) specimens at greater than 10(5) IU/ml, 15 of 16 (94%) at 10(4) to 10(5) IU/ml, 6 of 7 (86%) at 10(3) to 10(4) IU/ml, and 2 of 3 (67%) at less than 10(3) IU/ml. Quasispecies complexity, determined by using this novel long-amplicon method followed by heteroduplex mobility assay combined with single-stranded conformational polymorphism (HDA+SSCP) analysis, was very high, even during acute HCV infection, when 10 to 21 (median, 16) different HDA+SSCP patterns were detected among 33 cDNA clones examined. Replicate analyses indicate that this diversity is not dominated by random errors generated during amplification. Therefore, the HCV quasispecies is highly complex even during acute infection and is accurately represented in amplicons representing more than half of the viral genome. PMID- 15365016 TI - Use of the minimum spanning tree model for molecular epidemiological investigation of a nosocomial outbreak of hepatitis C virus infection. AB - The minimum spanning tree (MST) model was applied to identify the history of transmission of hepatitis C virus (HCV) infection in an outbreak involving five children attending a pediatric oncology-hematology outpatient ward between 1992 and 2000. We collected blood samples from all children attending since 1992, all household contacts, and one health care worker positive for antibody to HCV (anti HCV). HCV RNA detection was performed with these samples and with smears of routinely collected bone marrow samples. For all isolates, we performed sequence analysis and phylogenetic tree analysis of hypervariable region 1 of the E2 gene. The MST model was applied to clinical-epidemiological and molecular data. No additional cases were detected. All children, but not the health care worker, showed genotype 3a. On six occasions, all but one child had shared the medication room with another patient who later seroconverted. HCV RNA detection in bone marrow smears revealed, in some cases, a delay of several months in anti-HCV responses. Sequence analysis and phylogenetic tree analysis revealed a high identity among the isolates. The MST model applied to molecular data, together with the clinical-epidemiological data, allowed us to identify the source of the outbreak and the most probable patient-to-patient chain of transmission. The management of central venous catheters was suspected to be the probable route of transmission. In conclusion, the MST model, supported by an exhaustive clinical epidemiological investigation, appears to be a useful tool in tracing the history of transmission in outbreaks of HCV infection. PMID- 15365017 TI - Persistent histidine-rich protein 2, parasite lactate dehydrogenase, and panmalarial antigen reactivity after clearance of Plasmodium falciparum monoinfection. AB - We tested 240 patients with Plasmodium falciparum monoinfection for persistent parasite antigenemia after successful standardized antimalarial therapy by using the ICT Malaria Pf/Pv and OptiMAL-IT assays that detect the malaria antigens Plasmodium falciparum histidine-rich protein 2 (HRP2) and parasite lactate dehydrogenase (pLDH), respectively, as well as a panmalarial antigen (PMA). The patients were screened for antigenemia on days 0, 3, 7, and 14 of follow-up. On day 0, all 240 patients showed positive reactivity with both assays. Of the 229 cases with negative parasitemia on day 3, persistent antigenemia was observed in 207 (90.4%) of the cases: 188 (82.1%) for HRP2 antigen and 75 (32.8%) for PMA. There was a gradual decrease in antigenemia on follow-up to day 14; however, the drop in reactivity to PMA was less than that for HRP2 antigen. In contrast to HRP2 antigenemia, there was a significant decrease in pLDH antigenemia to 38.4% and to 14.8% (PMA) on day 3 (P < 0.03). The pLDH antigenemia level dropped further to 14.8% on day 7. There was no significant association of persistent antigenemia with gametocytemia. One case with gametocytemia was negative for both the antigens. In conclusion, the OptiMAL-IT assay is more sensitive than the ICT Malaria Pf/Pv test for monitoring therapeutic responses after antimalarial therapy since the LDH activity ceases when the malarial parasite dies. PMID- 15365018 TI - Risk factors for colonization and infection in a hospital outbreak caused by a strain of Klebsiella pneumoniae with reduced susceptibility to expanded-spectrum cephalosporins. AB - Between February 2001 and January 2002, an increase in the number of Klebsiella pneumoniae isolates with reduced susceptibility to expanded-spectrum cephalosporins (RSKp) was detected in the neonatal unit of the Juan Canalejo Hospital, and 21 patients were either colonized or infected by the bacterial isolates. The current "gold standard" method for typing K. pneumoniae isolates is pulsed-field gel electrophoresis. However, this technique is expensive and time consuming. In a search for faster and accurate alternatives to this method, we investigated PCR-based fingerprinting techniques (enterobacterial repetitive intergenic consensus sequence PCR [ERIC-PCR], repetitive extragenic palindromic sequence-based PCR [REP-PCR], and RAPD [randomly amplified polymorphic DNA]) for their ability to characterize K. pneumoniae isolates. The causal agent of the nosocomial outbreak was characterized by these techniques and was found to be a single epidemic strain (RSKp). A multiple regression logistic model was developed to identify potential independent factors associated with colonization and/or infection by RSKp. Logistic regression analysis was applied to all significant variables (P < 0.05) in the univariate analysis, and it was revealed that intubation (odds ratio [OR], 27.0; 95% confidence interval [95%CI], 5.39 to 135.14) and prematurity (OR, 4.4; 95%CI, 0.89 to 21.89) were such independent factors. Moreover, oxime cephalosporins did not appear to be statistically significant. Overall, the results showed that PCR-based techniques are expeditious and useful methods for typing K. pneumoniae isolates. Of the techniques studied, ERIC-PCR showed the highest discriminatory index (D = 0.828), followed by RAPD (D = 0.826) and REP-PCR (D = 0.773) PMID- 15365019 TI - Whole-blood counting immunoassay as a short-turnaround test for detection of hepatitis B surface antigen, anti-hepatitis C virus antibodies, and anti Treponema pallidum antibodies. AB - Whole-blood samples were used for a counting immunoassay (CIA) with the aim of developing a short- turnaround test. After optimization of the CIA, hepatitis B surface antigen (HBsAg), anti-hepatitis C virus antibodies (anti-HCV), and anti Treponema pallidum antibodies (anti-TP) were detected as efficiently as by an enzyme immunoassay (EIA) with serum samples. The correlations between whole-blood CIA and serum EIA were 99.8, 97.1, and 99.4% for HBsAg, anti-HCV, and anti-TP, respectively. Whole-blood CIA may be of value when rapid screening of many samples is required. PMID- 15365020 TI - Identification and typing of Malassezia species by amplified fragment length polymorphism and sequence analyses of the internal transcribed spacer and large subunit regions of ribosomal DNA. AB - Malassezia yeasts are associated with several dermatological disorders. The conventional identification of Malassezia species by phenotypic methods is complicated and time-consuming, and the results based on culture methods are difficult to interpret. A comparative molecular approach based on the use of three molecular techniques, namely, amplified fragment length polymorphism (AFLP) analysis, sequencing of the internal transcribed spacer, and sequencing of the D1 and D2 domains of the large-subunit ribosomal DNA region, was applied for the identification of Malassezia species. All species could be correctly identified by means of these methods. The results of AFLP analysis and sequencing were in complete agreement with each other. However, some discrepancies were noted when the molecular methods were compared with the phenotypic method of identification. Specific genotypes were distinguished within a collection of Malassezia furfur isolates from Canadian sources. AFLP analysis revealed significant geographical differences between the North American and European M. furfur strains. PMID- 15365021 TI - Simple subtyping assay for human immunodeficiency virus type 1 subtypes B, C, CRF01-AE, CRF07-BC, and CRF08-BC. AB - After the initial development of a human immunodeficiency virus type 1 (HIV-1) subtype-screening tool by nested multiplex PCR, we further improve it through the redesign of subtype-specific primers based on subtype signature pattern (SSP) analyses and optimization of the PCR conditions. Extracted RNA from plasma samples was used in reverse transcription and the cDNA products were added to the first round PCR, in which universal primers in the gag region were used to detect HIV-1 M group isolates. In the second round of PCR, three pairs of subtype specific primers, detecting subtypes B, C, and CRF01-AE, were added in one tube. Subtype determination was based on the different size of PCR products on the agarose gel electrophoresis. An additional set of primers detecting only the prevalent recombinant strains CRF07-BC and CRF08-BC was used to discriminate CRF07- and CRF08-BC from pure subtype C. Testing for all kinds of HIV subtype reference strains indicated that this assay was applicable. A panel of 252 HIV positive samples and 30 HIV-negative samples was further used to evaluate and validate this assay. Compared to the assay of sequence-based phylogenetic analysis, the newly developed assay has an adequate designated subtype sensitivity, 93.2% (69 of 74) for subtype B, 95.1% (117 of 123) for subtype C, 94.0% (47 of 50) for CRF01-AE, and 95.0% (115 of 121) for CRF07-BC and CRF08-BC. Most importantly, the intersubtype specificity of the assay was found to be 100%. The assay specificity was also found to be 100% when used to test 30 HIV-negative samples. The average reproducibility was 96.0% for subtype B, 96.7% for subtype C, and 95.0% for CRF01-AE. We have developed a simple, rapid, and low cost assay for screening subtypes B, C, CRF01-AE, CRF07-BC, and CRF08-BC in China. PMID- 15365022 TI - Use of an oligonucleotide array for laboratory diagnosis of bacteria responsible for acute upper respiratory infections. AB - We developed a diagnostic array of oligonucleotide probes targeting species specific variable regions of the genes encoding topoisomerases GyrB and ParE of respiratory bacterial pathogens. Suitable broad-range primer sequences were designed based on alignment of gyrB/parE sequences from nine different bacterial species. These species included Corynebacterium diphtheriae, Fusobacterium necrophorum, Haemophilus influenzae, Legionella pneumophila, Moraxella catarrhalis, Mycoplasma pneumoniae, Staphylococcus aureus, Streptococcus pneumoniae, and Streptococcus pyogenes. Specific probe sequences were selected by comparative analysis against the European Bioinformatics Database, as well as gyrB/parE sequences generated for this study. To verify specificity, at least six initial oligonucleotide probe sequences per bacterial species were tested by hybridization on a solid glass support using culture collection strains as templates. Finally, three oligonucleotide probes per bacterial species were utilized to examine 65 middle ear fluid and 29 throat swab samples. The sensitivities of the developed assay compared to classic culture from middle ear fluid samples for H. influenzae, M. catarrhalis, and S. pneumoniae were 96 (93 for culture), 73 (93 for culture), and 100% (78% for culture), respectively. No cross-reactivity with bacterial species belonging to the normal oral flora was observed when the 29 throat swab samples were studied. The sensitivity of the assay to detect S. pyogenes from these samples was 93% (80% for culture). These results provide a proof of concept for the diagnostic use of microarray technology based on broad-range topoisomerase gene amplification, followed by hybridization and specific detection of bacterial species. PMID- 15365023 TI - Uses of Staphylococcus aureus GeneChips in genotyping and genetic composition analysis. AB - Understanding the relatedness of strains within a bacterial species is essential for monitoring reservoirs of antimicrobial resistance and for epidemiological studies. Pulsed-field gel electrophoresis (PFGE), ribotyping, and multilocus sequence typing are commonly used for this purpose. However, these techniques are either nonquantitative or provide only a limited estimation of strain relatedness. Moreover, they cannot extensively define the genes that constitute an organism. In the present study, 21 oxacillin-resistant Staphylococcus aureus (ORSA) isolates, representing eight major ORSA lineages, and each of the seven strains for which the complete genomic sequence is publicly available were genotyped using a novel GeneChip-based approach. Strains were also subjected to PFGE and ribotyping analysis. GeneChip results provided a higher level of discrimination among isolates than either ribotyping or PFGE, although strain clustering was similar among the three techniques. In addition, GeneChip signal intensity cutoff values were empirically determined to provide extensive data on the genetic composition of each isolate analyzed. Using this technology it was shown that strains could be examined for each element represented on the GeneChip, including virulence factors, antimicrobial resistance determinants, and agr type. These results were validated by PCR, growth on selective media, and detailed in silico analysis of each of the sequenced genomes. Collectively, this work demonstrates that GeneChips provide extensive genotyping information for S. aureus strains and may play a major role in epidemiological studies in the future where correlating genes with particular disease phenotypes is critical. PMID- 15365024 TI - Rapid and specific detection of tdh, trh1, and trh2 mRNA of Vibrio parahaemolyticus by transcription-reverse transcription concerted reaction with an automated system. AB - Vibrio parahaemolyticus strains carrying the thermostable direct hemolysin (TDH) tdh gene, the TDH-related hemolysin (trh) gene, or both genes are considered virulent strains. We previously demonstrated that the transcription-reverse transcription concerted (TRC) method could be used to quantify the amount of mRNA transcribed from the tdh gene by using an automated detection system. In this study, we devised two TRC-based assays to quantify the mRNAs transcribed from the trh1 and trh2 genes, the two representative trh genes. The TRC-based detection assays for the tdh, trh1, and trh2 transcripts could specifically and quantitatively detect 10(3) to 10(7) copies of the corresponding calibrator RNAs. We examined by the three TRC assays the total RNA preparations extracted from 103 strains of Vibrio parahaemolyticus carrying the tdh, trh1, or trh2 gene in various combinations. The tdh, trh1, and trh2 mRNAs in the total RNA preparations were specifically quantified, and the time needed for detection ranged from 9 to 19 min, from 14 to 18 min, and from 9 to 12 min, respectively. The results showed that this automated TRC assays could detect the tdh, trh1, and trh2 mRNAs specifically, quantitatively, and rapidly. The relative levels of TDH determined by the immunological method and that of tdh mRNA determined by the TRC assays for most tdh-positive strains correlated. Interestingly, the levels of TDH produced from the strains carrying both tdh and trh genes were lower than those carrying only the tdh gene, whereas the levels of mRNA did not significantly differ between the two groups. PMID- 15365025 TI - Simple chemical extraction method for DNA isolation from Aspergillus fumigatus and other Aspergillus species. AB - DNA was successfully isolated from numerous Aspergillus spp. by use of a commercial kit. DNA that was easily digested and yielded PCR products up to 8.5 kb in size was recovered from broth or agar cultures. The ease and speed of this protocol provide an alternative to physical methods of DNA isolation. PMID- 15365026 TI - Surveillance study (2000 to 2001) of G- and P-type human rotaviruses circulating in South Korea. AB - Human rotavirus VP4 and VP7 gene sequences were amplified by reverse transcription-PCR from 53% (322 of 607) of fecal specimens collected from children with severe diarrhea who visited hospitals in six urban areas of South Korea in 2000 and 2001. G2 was the most frequently found G type (constituted 50.6%), followed by G1 (30.1%) and G4 (13.0%). Although the P types of high incidence were P[4] (53.1%) and P[8] (21.4%), a significant incidence of P[6] (20.2%) was also noticeable. The commonest G- and P-type combination found in this study was G2P[4], rather than G1P[8], the most prevalent type known worldwide. PMID- 15365027 TI - Potential screening assay for undetectable viruses on the basis of their capacity to induce alpha interferon production. AB - High interferon production by plasmacytoid dendritic cells (PDC) was unexpectedly noted after their coculture with CD4(+) cells from a healthy donor whose cells subsequently showed human herpesvirus type 6 and 7 infections. This release of interferon was not observed with uninfected normal CD4(+) cells. Induction of PDC interferon production could help screen for covert virus infections. PMID- 15365028 TI - Usefulness of the GenoType MTBC assay for differentiating species of the Mycobacterium tuberculosis complex in cultures obtained from clinical specimens. AB - A novel DNA strip assay, GenoType MTBC, was evaluated for differentiation of Mycobacterium tuberculosis complex species from 77 positive liquid cultures in clinical practice. Species identification (M. tuberculosis [71 strains], Mycobacterium bovis subsp. bovis [5 strains], and Mycobacterium africanum subtype I [1 strain]) results were identical to conventional results. The sensitivity was slightly higher for this test than for the AccuProbe assay. PMID- 15365029 TI - Comparison of the real-time PCR method and the Gen-Probe amplified Mycobacterium tuberculosis direct test for detection of Mycobacterium tuberculosis in pulmonary and nonpulmonary specimens. AB - Real-time PCR was compared to Amplified Mycobacterium tuberculosis Direct Test (AMTDII) for 100 clinical specimens. The overall sensitivities of the real-time PCR method and AMTDII were similar for respiratory and nonrespiratory specimens. However, real-time PCR seemed to be less susceptible to amplification inhibitors than AMTDII. PMID- 15365030 TI - Fosfomycin and rifampin disk diffusion tests for detection of Escherichia coli mutator strains. AB - A simple method, using commercial disks to detect Escherichia coli mutator strains, is proposed. The breakpoint for detecting strains with a mutation frequency >/=5 x 10(-7) was established at >/=70 and >/=20 colonies in the inhibition zone of fosfomycin and rifampin disks, respectively, after seeding 100 microl of an overnight culture. Strains with <30 and <10 colonies in fosfomycin and rifampin inhibition zones are presumptively non-mutators. PMID- 15365031 TI - Multiplex PCR method for identifying recombinant vaccine-related polioviruses. AB - The recent discovery of recombinant circulating vaccine-derived poliovirus (recombinant cVDPV) has highlighted the need for enhanced global poliovirus surveillance to assure timely detection of any future cVDPV outbreaks. Six pairs of Sabin strain-specific recombinant primers were designed to permit rapid screening for VDPV recombinants by PCR. PMID- 15365032 TI - Community-acquired Clostridium difficile diarrhea caused by binary toxin, toxin A, and toxin B gene-positive isolates in Hungary. AB - The aim of this work was to study the toxin types of Clostridium difficile isolates originating from different parts of Hungary. A PCR method was used for amplification of the two major toxin genes and the binary toxin gene and to detect the deletion or insertion in the 3' end of the toxin A gene. The findings were compared with the results of cytotoxicity assays on the HeLa cell line. One hundred twelve isolates were tested; the toxin A and toxin B genes were detected in 79 strains by the PCR method. All of the isolates that were positive by the PCR method were also positive by the cytotoxicity assay. All of the other strains (n = 33) were negative for the toxin A and toxin B genes; in these cases, cytopathic effects on the cell line were not observed. No tcdA-negative and tcdB positive isolates were found by the PCR method. In two cases, the presence of a binary toxin gene was observed by PCR; both isolates that were isolated from diarrheal feces carried the tcdA and tcdB genes. No prior hospitalization had occurred in either case. PMID- 15365033 TI - In vitro susceptibilities of isolates of Sporothrix schenckii to itraconazole and terbinafine. AB - Thirty isolates of the yeast form of Sporothrix schenckii were evaluated for in vitro susceptibility to itraconazole and terbinafine by the recommended NCCLS modified technique (M27-A2). The MICs of itraconazole obtained oscillated between 0.062 and 4.0 microg/ml, and those of terbinafine oscillated between 0.007 and 0.50 microg/ml; therefore, terbinafine showed greater in vitro activity. PMID- 15365034 TI - Prevalence of hepatitis C virus infection among hemodialysis patients at a tertiary-care hospital in Mexico City, Mexico. AB - We determined the prevalence of hepatitis C virus (HCV) in hemodialysis patients by antibody testing and HCV RNA determination by PCR. A total of 149 patients with kidney failure with replacement therapy were tested. The prevalence of anti HCV was 6.7% (10 of 149 patients), and viremia was detectable in 8 of 149 (5%) patients. Three of 149 patients (2%) were anti-HCV negative with detectable HCV RNA. PMID- 15365035 TI - Antimicrobial susceptibilities of invasive pediatric Abiotrophia and Granulicatella isolates. AB - Abiotrophia and Granulicatella species have been associated with various infections. Antimicrobial susceptibility data for these nutritionally variant streptococcus-like organisms, especially for pediatric isolates, are very limited. Little is known about the genetic bases of their resistance mechanisms. We report the results of identification to bacterial species level, antimicrobial susceptibility testing, macrolide resistance testing, and detection of genes encoding that resistance for a collection of 15 pediatric clinical isolates from normally sterile sites. Our results indicate that the prevalence of beta-lactam and macrolide resistance is high and that both erm and mef are found in these isolates. PMID- 15365036 TI - Evaluation of conventional castaneda and lysis centrifugation blood culture techniques for diagnosis of human brucellosis. AB - We investigated the role of the lysis centrifugation blood culture technique over the conventional Castaneda technique for the diagnosis of human brucellosis. The lysis centrifugation technique has been found to be more sensitive in both acute (20% higher sensitivity; P < 0.00001) and chronic (40% higher sensitivity; P = 0.087) forms of brucellosis. The major advantage of lysis centrifugation was in the mean detection time, which was only 2.4 days in acute and 2.7 days in chronic cases, with 103 out of 110 (93.6%) and 17 out of 20 (85%) cultures from acute and chronic brucellosis, respectively, detected before the conventional culture was positive. Our results confirmed the potential usefulness of the lysis technique in diagnosis and institution of appropriate antibiotic therapy. PMID- 15365037 TI - Detection of hepatitis A virus RNA in saliva. AB - Hepatitis A virus (HAV) is shed in feces but also in saliva. HAV RNA was detected in saliva in five out of six acutely infected patients with HAV viremia. Serum and saliva sequences were identical. The simplicity of obtaining material allows the recommendation of the use of saliva for investigation of outbreaks. PMID- 15365038 TI - Multicenter validation of the cppB gene as a PCR target for detection of Neisseria gonorrhoeae. AB - The cppB gene is often used as a target for detection of Neisseria gonorrhoeae by PCR. Using a coded panel of 500 DNA samples, we determined that the cppB gene is missing in 5.8% of N. gonorrhoeae strains, and therefore we consider the cppB gene to be an unsuitable target. PMID- 15365039 TI - Variability of germinative potential among pathogenic species of Aspergillus. AB - The objective of our study was to evaluate parameters influencing the germination of Aspergillus conidia. Inoculum concentration and age significantly influenced germination. Different incubation temperatures revealed significant differences among Aspergillus species. The internal human milieu provides the ideal conditions for the development of invasive disease by Aspergillus fumigatus but restricts invasion by Aspergillus flavus and Aspergillus niger. PMID- 15365040 TI - Sequencing and phylogenetic analysis of human genotype P[6] rotavirus strains detected in Hungary provides evidence for genetic heterogeneity within the P[6] VP4 gene. AB - Although rotavirus genotype P[6] is one of the three most common VP4 specificities associated with human infection, the relatively few sequence data available in public databases suggest that the genetic variability within P[6] might be presently unexplored. Thus far, two human P[6] lineages (M37-like and AU19-like) and a single porcine P[6] lineage (Gottfried-like) have been identified by phylogenetic analysis. Serologic studies demonstrated that these three lineages are antigenically distinct from each other, a finding based on which they were classified into three subtypes, P2A[6] (M37-like), P2B[6] (Gottfried-like), and P2C[6] (AU19-like). To study heterogeneity within this genotype, we selected for molecular characterization a total of six P[6] strains detected during an ongoing surveillance in Hungary. The variable region of the VP4 gene was subjected to sequencing and phylogenetic analysis. Our data indicated that these six strains fell into two phylogenetic lineages distinguishable from the human lineages M37-like and AU19-like and from the porcine lineage Gottfried-like. Further studies are needed to understand whether these two novel lineages are genuine human strains or might have originated from animal strains and to evaluate the antigenic relationship of the novel Hungarian P[6] strains to the three established subtypes. PMID- 15365041 TI - Detection and identification of Actinobacillus pleuropneumoniae serotypes 1, 2, and 8 by multiplex PCR. AB - Multiplex PCR assays were developed to identify Actinobacillus pleuropneumoniae serotypes 1, 2, and 8. Primers designed for the conserved capsular polysaccharide (CP) export region amplified a 489-bp DNA fragment from all serotypes. Primers specific to the CP biosynthesis regions of serotypes 1, 2, and 8 amplified fragments of 1.6 kb, 1.7 kb, and 970 bp from only their respective serotypes. PMID- 15365043 TI - Pneumolysin is a key factor in misidentification of macrolide-resistant Streptococcus pneumoniae and is a putative virulence factor of S. mitis and other streptococci. AB - We evaluated the applicability of ply PCR for confirmation of the identification of Streptococcus pneumoniae. lytA PCR, 16S rRNA sequencing, and amplified fragment length polymorphism were used as reference methods. In contrast to the lytA gene, the ply gene proved to be not specific for S. pneumoniae. The presence of the ply gene in other streptococci, in particular Streptococcus mitis, suggests that pneumolysin plays a pathogenic role. PMID- 15365042 TI - Characterization of avian H3N3 and H1N1 influenza A viruses isolated from pigs in Canada. AB - H3N3 and H1N1 influenza A viruses were isolated from Canadian pigs in 2001 and 2002. These viruses are phylogenetically related to waterfowl viruses and antigenically distinct from reference swine influenza viruses. The isolation of these viruses reemphasizes the potential for interspecies transmission of influenza viruses from waterfowl to pigs in North America. PMID- 15365044 TI - Comparison of the Sensititre YeastOne colorimetric antifungal panel with a modified NCCLS M38-A method to determine the activity of voriconazole against clinical isolates of Aspergillus spp. AB - The susceptibilities of 63 isolates of Aspergillus spp. to voriconazole were evaluated by a modified NCCLS M38-A method and the Sensititre YeastOne method. The overall agreement was 82.5%, ranging from 100% for Aspergillus niger and Aspergillus terreus to 62.5% for Aspergillus flavus. Discrepancies between the methods were due to higher Sensititre MICs. The Sensititre YeastOne method could have potential value for susceptibility testing of Aspergillus spp. to voriconazole. PMID- 15365045 TI - Development of a real-time PCR assay using SYBR Green I for provirus load quantification in a murine model of AIDS. AB - A real-time PCR assay using SYBR Green I for quantification of provirus load in a murine model of AIDS (i.e., LP-BM5 infection) was developed and validated. In this method, data are normalized against the 18S rRNA gene. The method has a dynamic range of 8 logs and a sensitivity of one copy. PMID- 15365047 TI - Pathogenic differences between North American and Latin American strains of Histoplasma capsulatum var. capsulatum in experimentally infected mice. AB - Clinical differences in histoplasmosis between North America and Brazil prompted investigation of experimental infection with representative strains. Mortality was higher with Latin American strains, and lung pathology showed large necrotizing granuloma with prominent neutrophilic infiltration. Chronic disease was unique to the North American strain. PMID- 15365046 TI - Differing patterns of liver disease progression and hepatitis C virus (HCV) quasispecies evolution in children vertically coinfected with HCV and human immunodeficiency virus type 1. AB - Hepatitis C virus (HCV) quasispeciation was studied in two children vertically coinfected with HCV and human immunodeficiency virus type 1 (HIV-1). HCV quasispecies diversification and liver injury were more significant in patient C1, who was immunocompetent with anti-HIV therapy, than in patient C2, who was immunosuppressed, in consistency with modulation of HCV quasispeciation and liver injury by immunocompetence in coinfected children. PMID- 15365048 TI - Cladophialophora bantiana brain abscess in a solid-organ transplant recipient: case report and review of the literature. AB - Cerebral phaeohyphomycosis caused by Cladophialophora bantiana is a rare disease. We describe a heart and bilateral lung transplant recipient who was unsuccessfully treated for a C. bantiana brain abscess. This report compares the present case to those of other solid-organ transplant recipients with the same infection and to those of patients who did not receive transplants. PMID- 15365049 TI - Neonatal Candida albicans septic thrombosis of the portal vein followed by cavernous transformation of the vessel. AB - We report two premature neonates with Candida albicans septic thrombosis of the portal vein who developed, in very early childhood, the sonographic appearance of cavernous transformation of the vessel and/or clinical signs of extrahepatic portal hypertension. PMID- 15365050 TI - Pyelonephritis and urosepsis caused by Streptococcus pneumoniae. AB - This report presents the case of a patient with a massive pyelonephritis and a urosepsis caused by Streptococcus pneumoniae. This case is unusual as the focus was distant from the respiratory tract, the usual primary site of infection caused by this organism. No other primary site of infection was documented. PMID- 15365052 TI - Bacteremia caused by Clostridium symbiosum. AB - We describe a fatal case of Clostridium symbiosum bacteremia in a 70-year-old man with metastatic colon cancer. Our report is the first, in the world literature, of human infection caused by this microorganism. PMID- 15365051 TI - Mycobacterium heckeshornense infection in an immunocompetent patient and identification by 16S rRNA sequence analysis of culture material and a histopathology tissue specimen. AB - Mycobacterium heckeshornense is a rare isolate in clinical specimens. We performed simultaneous 16S rRNA sequence analysis of a mycobacterium culture and a histopathology specimen to determine the relevance of M. heckeshornense infection in an immunocompetent patient initially presenting with pneumothorax. PMID- 15365053 TI - Bacteremic infection with Pantoea ananatis. AB - A 73-year-old man was hospitalized for dyspnea and bilateral ankle edema. During his hospital stay he presented anal hemorrhage and developed a high fever after colonoscopy. A set of aerobic and anaerobic blood culture bottles yielded a pure culture of gram-negative rods, susceptible to all antibiotics tested. The API20E code was 1005133, resulting in a very good identification as Pantoea sp. Subsequent sequencing of the 16S rRNA gene revealed a final identification as Pantoea ananatis. The patient was given intravenous and oral therapy with piperacillin-tazobactam and ofloxacin and recovered completely from his infection. PMID- 15365054 TI - Streptococcus porcinus as a cause of spontaneous preterm human stillbirth. AB - We report, to our knowledge, on the first case of a woman suffering stillbirth due to Streptococcus porcinus on the basis of microbiologic and histologic data. PMID- 15365055 TI - Continuous ambulatory peritoneal dialysis-related peritonitis associated with Lancefield group G beta-hemolytic streptococcus: report of two cases requiring Tenckhoff catheter removal. AB - We describe the first two cases of continuous ambulatory peritoneal dialysis related peritonitis associated with Lancefield group G beta-hemolytic streptococci in the literature. Both patients presented with abdominal pain and turbid dialysis effluent with or without fever. Both had concomitant gastrointestinal tract disturbance. Both did not respond to intraperitoneal cefazolin and tobramycin and required removal of the Tenckhoff catheters. PMID- 15365056 TI - Rapid identification of Mycobacterium genavense with a new commercially available molecular test, INNO-LiPA MYCOBACTERIA v2. AB - We report a rare mesenteric localized Mycobacterium genavense infection in a severely immunocompromised human immunodeficiency virus-infected patient. An INNO LiPA MYCOBACTERIA v2 test was performed directly on biopsy samples. This new molecular tool could be used for simultaneous identification of mycobacterium species from human specimens, but other studies are needed to validate our first results. PMID- 15365057 TI - Campylobacter fetus of reptile origin as a human pathogen. AB - A Campylobacter species was isolated from blood from a febrile patient with precursor T-cell acute lymphoblastic leukemia, and after antibiotic treatment, a similar bacterium was isolated from blood 37 days later. Although phenotypic testing did not definitively identify the organisms, molecular analysis indicated that they were the same strain of Campylobacter fetus subsp. fetus and were of reptile origin. PMID- 15365058 TI - Cost-effective method for identification of dimorphic fungi. AB - Traditional methods to identify dimorphic fungi dictate that the mold be converted to the yeast phase at 35 to 37 degrees C. We present a time- and cost saving method of confirming the identification of a dimorph by using special stains to demonstrate the yeast phase directly growing in the original clinical specimens. PMID- 15365059 TI - Centrifugal ultrafiltration method for rapid concentration of Legionella pneumophila urinary antigen. PMID- 15365060 TI - False-positive result provided by rapid heterophile antibody test in a case of acute infection with hepatitis E virus. PMID- 15365061 TI - Cefoxitin does not induce production of penicillin binding protein 2a in methicillin-susceptible Staphylococcus aureus strains. PMID- 15365062 TI - Considerations in evaluating the applicability of universal detection of oral pathogens. PMID- 15365063 TI - Shiga toxin antigen detection should not replace sorbitol MacConkey agar screening of stool specimens. PMID- 15365064 TI - Factors predictive of tumor-positive nonsentinel lymph nodes after tumor-positive sentinel lymph node dissection for melanoma. AB - PURPOSE: Approximately 20% of sentinel node (SN) positive melanoma patients have additional non-SN (NSN) metastasis. The rationale for this study was to identify the factors associated with additional nodal disease, as a method to determine which patients may most benefit from completion lymph node dissection (CLND). PATIENTS AND METHODS: During 1990 to 2002, 1,599 patients have undergone SN biopsy at our institute. 19.5% underwent CLND for tumor-positive SN. One hundred ninety-one of these patients had clinicopathologic information available for review. Univariate analyses used chi2 test, Wilcoxson rank sum test, and chi2 test for trend. Multivariate analyses used logistic regression and Wald test. RESULTS: Forty-six (24%) patients had tumor-positive NSN. Univariate analyses showed that primary thickness (Breslow and Clark), primary site, SN tumor size, and number of tumor-positive SNs were significantly associated with tumor positive NSN. Multivariate analysis (167 patients), confirmed that Breslow and SN tumor size were independently predictive. Sex, histology, ulceration, mitotic index, and SN basin location were not predictive. Risk stratification by the number of prognostic factors present (Breslow > or = 3 mm and SN tumor size > or = 2 mm) showed that probability of finding tumor-positive NSN was 12.3% in the low-risk group (0 factors), 30.9% in the intermediate-risk group (1 factor), and 41.9% in the high-risk group (2 factors). CONCLUSION: Thicker primary and larger SN tumor size are factors that correlate best with tumor-positive NSN. Although none of these factors are absolutely predictive of residual nodal disease, these factors must be strongly considered if the SN contains metastasis, as they provide enhanced risk assessment for NSN tumor-positivity. PMID- 15365065 TI - Bone mineral density among premenopausal women with early breast cancer in a randomized trial of adjuvant endocrine therapy. AB - PURPOSE: To examine the effects on bone mineral density of 2 years of treatment with a luteinizing hormone-releasing hormone (LHRH) agonist alone or in combination with tamoxifen or tamoxifen alone in premenopausal breast cancer. PATIENTS AND METHODS: We recruited 89 women from two centers in Stockholm participating in a randomized multicenter trial of three different endocrine approaches in the adjuvant setting (Zoladex in Premenopausal Patients Trial). The women were assigned to receive the LHRH agonist goserelin with or without tamoxifen, tamoxifen alone, or no endocrine therapy. The treatment was given for 2 years. We measured total-body bone density before start of treatment and at 12, 24, and 36 months. RESULTS: After 2 years of treatment, there was a significant loss of bone mineral density (mean change, -5%; P <.001) in the women receiving goserelin alone. The combined goserelin and tamoxifen treatment, as well as tamoxifen alone, resulted in a lesser but statistically significant decline in bone mineral density (mean change, -1.4%; P =.02; and -1.5%; P <.001). One year after cessation of treatment, the goserelin group alone showed a partial recovery from bone loss (mean change, 1.5%; P =.02). CONCLUSION: Two years of ovarian ablation from goserelin treatment caused a significant reduction in bone mineral density but there was a partial recovery from the bone loss 1 year after cessation of treatment. The addition of tamoxifen seems to partially counteract the demineralizing effects of goserelin. PMID- 15365066 TI - Effect of doxorubicin plus cyclophosphamide on left ventricular ejection fraction in patients with breast cancer in the North Central Cancer Treatment Group N9831 Intergroup Adjuvant Trial. AB - PURPOSE: To evaluate changes in left ventricular ejection fraction (LVEF) after four cycles of adjuvant doxorubicin plus cyclophosphamide (AC) in women with human epidermal growth factor receptor 2-positive (node-positive or node negative) breast cancer enrolled onto the North Central Cancer Treatment Group N9831 Intergroup Adjuvant Trial. PATIENTS AND METHODS: Patients were randomly assigned to receive standard doxorubicin (60 mg/m2) plus cyclophosphamide (600 mg/m2) every 3 weeks for four cycles followed by (1) weekly paclitaxel for 12 weeks; (2) weekly paclitaxel for 12 weeks, then weekly trastuzumab for 52 weeks; or (3) weekly paclitaxel plus trastuzumab for 12 weeks, then weekly trastuzumab for 40 weeks. LVEF was monitored before and after AC. RESULTS: Of the 1,576 eligible patients who completed AC, 1,458 had pre- and post-AC LVEF measurements taken using the same methodology (multiple-gated acquisition in 1,153 patients and echocardiogram in 305 patients). Among these 1,458 patients, 745 (51.1%) had < or = 15% decrease in LVEF and LVEF that remained at or above the radiologic lower limit of normal (LLN); 42 patients (2.9%) had < or = 15% decrease in LVEF and LVEF that decreased to or below the LLN; and 37 patients (2.5%) had an LVEF decrease of more than 15%. There was grade 2 LVEF toxicity in 96 (6.6%) of the 1,458 patients. CONCLUSION: Standard AC chemotherapy is associated with frequent decreases in LVEF, which are noted when measured 3 weeks after completion of the fourth cycle. Patients are being observed to determine the long-term significance of this and the potential impact on subsequent treatment options. PMID- 15365067 TI - Residual tumor resection after high-dose chemotherapy in patients with relapsed or refractory germ cell cancer. AB - PURPOSE: To assess the role of residual tumor resection performed after high-dose chemotherapy (HDCT) in patients with relapsed or refractory germ cell tumors (GCT). PATIENTS AND METHODS: Between July 1987 and October 1999, postchemotherapy resections of residual tumors were performed in 57 patients who had been treated with HDCT for relapsed or refractory GCT and who had achieved a partial remission to this treatment. RESULTS: Complete resections of residual masses were achieved in 52 (91%) of 57 patients who were rendered disease free; in five (9%) of 57 patients, the resections were incomplete. Resection of a single site was performed in 39 (68%) of 57 patients, and the remaining 18 (32%) of 57 patients required interventions at two or more residual tumor sites. Necrosis was found in 22 (38%) of 57 patients, mature teratoma with or without necrosis was found in nine (16%) of 57 patients, and viable cancer with or without additional necrosis or mature teratoma was found in 26 (46%) of 57 patients. Viable cancer consisted either of residual germ cell or undifferentiated cancer in 22 (85%) of 26 patients, with additional non-GCT histologies in the remaining four patients. Patients with viable cancer had a significantly inferior outcome after surgery compared with patients with necrosis and/or mature teratoma even if all cancer was completely resected. Pulmonary lesions with a diameter of more than 2 cm were the only predictive variable for viable cancer in univariate analysis. CONCLUSION: Resections of all residual tumors should be attempted in patients with relapsed or refractory GCT and partial remissions after HDCT. PMID- 15365068 TI - Phase II trial of bortezomib for patients with advanced renal cell carcinoma. AB - PURPOSE: To assess the efficacy and toxicity of bortezomib (Velcade; Milennium Pharmaceuticals Inc, Cambridge, MA; formerly PS-341) in patients with metastatic renal cell carcinoma (RCC). PATIENTS AND METHODS: Thirty-seven patients with metastatic RCC were treated with bortezomib. The first 25 patients enrolled onto the trial were treated with a dose of 1.5 mg/m2. The dose was decreased to 1.3 mg/m2 for the subsequent 12 patients, because more than 50% of the patients treated at the higher dose required dose reductions. Bortezomib was given by intravenous administration on a twice-weekly schedule for 2 weeks followed by 1 week without treatment until progression or unacceptable toxicity occurred. Twenty-three patients (62%) previously had undergone nephrectomy, and 19 patients (51%) had previously been treated with cytokine therapy. RESULTS: Of the 37 assessable patients, the best response was a partial response in four patients (11%; 95% CI, 3% to 25%) and stable disease in 14 patients (38%; 95% CI, 23% to 55%). The four patients with partial response experienced response durations of 8, 8+, 15+, and 20+ months. Grade 2 or 3 sensory neuropathy was present in 10 patients (53%) overall. One patient in the 1.5 mg/m2 group had grade 3 sensory neuropathy; no grade 3 sensory neuropathy was seen in the 1.3 mg/m2 group. CONCLUSION: The results of this trial suggest that bortezomib has an antitumor effect in individual patients with metastatic RCC. The small proportion of patients who achieved a partial response does not support routine use in metastatic RCC. Efforts to identify the molecular profile associated with clinical response or combination therapy with interferon alfa or other novel agents, may be considered. PMID- 15365069 TI - Impact of the percentage of positive prostate cores on prostate cancer-specific mortality for patients with low or favorable intermediate-risk disease. AB - PURPOSE: We investigated whether pretreatment factors predicted time to prostate cancer-specific mortality (PCSM) after conventional-dose and conformal radiation therapy (CRT). PATIENTS AND METHODS: Between 1988 and 2002, 421 patients with low (prostate-specific antigen [PSA] level < or = 10 ng/mL and biopsy Gleason score < or = 6) or favorable intermediate-risk (PSA > 10 to 15 ng/mL or biopsy Gleason score 3 + 4, but not both factors) disease underwent CRT (median dose, 70.4 Gy). Cox regression multivariable analysis was performed to determine whether the PSA level, Gleason score, T category, or the percentage of positive cores (% PC) predicted time to PCSM after CRT. After a median follow-up of 4.5 years, 117 (28%) patients have died. RESULTS: The % PC was the only significant predictor (Cox P < or =.03). The relative risk of PCSM after CRT for patients with > or = 50% as compared with less than 50% PC was 10.4 (95% CI, 1.2 to 87; Cox P =.03), 6.1 (95% CI, 1.3 to 28.6; Cox P =.02), and 12.5 (95% CI, 1.5 to 107; Cox P =.02) in men with a PSA < or = 10 and Gleason score < or = 6, PSA < or = 10 and Gleason score < or = 7, and PSA < or = 15 and Gleason score < or = 6, respectively. By 5 years after CRT, 5% to 9% compared with less than 1% (log-rank P < or =.01) of these patients experienced PCSM if they had > or = 50% compared with less than 50% PC, respectively. CONCLUSION: CRT dose-escalation techniques, the addition of hormonal therapy, or both should be considered in the management of patients with low or favorable intermediate-risk disease and > or = 50% PC. PMID- 15365070 TI - Randomized comparison of power Doppler ultrasound-directed excisional biopsy with standard excisional biopsy for the characterization of lymphadenopathies in patients with suspected lymphoma. AB - PURPOSE: The sensitivity of lymph node excisional biopsy requires validation. Power Doppler ultrasound (US) helps predict the malignant status of lymphadenopathies. We used power Doppler US to select for biopsy the lymph node most suspected of malignancy. PATIENTS AND METHODS: One hundred fifty-two patients having lymphadenopathies with clinical suspicion of lymphoma were divided into two well-matched groups and randomly assigned to undergo either standard or power Doppler US-directed lymph node excisional biopsy. RESULTS: Histology showed a malignancy in 64% of patients in the standard group (lymphoma, 49 patients; carcinoma, two patients) and in 87% of patients in the US-assisted group (lymphoma, 62 patients; carcinoma, one patient). There were significantly fewer biopsy-related complications in the assisted group than in the standard group. During the follow-up of the patients with lymph nodes reported as being reactive, 14 of 29 patients in the standard group were rebiopsied and were found to have lymphoma (13 patients) or carcinoma at the subsequent lymph node histology, whereas none of the patients in the assisted group (nine patients) required a second biopsy. Thus, biopsy provided false-negative results for malignancy in 21% of patients affected by lymphoma in the standard group and never in the assisted group (P <.01). CONCLUSION: Power Doppler US is an accurate tool for screening lymphadenopathies to be removed by excisional biopsy in patients with suspected lymphoma. PMID- 15365071 TI - Serum cardiac troponins and N-terminal pro-brain natriuretic peptide: a staging system for primary systemic amyloidosis. AB - PURPOSE: Primary systemic amyloidosis (AL) is a multisystemic disorder resulting from an underlying plasma cell dyscrasia. There is no formal staging system for AL, making comparisons between studies and treatment centers difficult. Our group previously identified elevated serum cardiac troponin T (cTnT) as the most powerful predictor of overall survival. Others have reported that N-terminal pro brain natriuretic peptide (NT-proBNP) is a valuable prognostic marker. We sought to develop a staging system for patients with AL. PATIENTS AND METHODS: Two hundred forty-two patients with newly diagnosed AL who were seen at the Mayo Clinic between April 1979 and November 2000, and who had echocardiograms and stored serum samples at presentation were eligible for this retrospective review. NT-proBNP measurements were performed on 242 patients in whom cTnT and cardiac troponin I (cTnI) had been previously run. Two prognostic models were designed using threshold values of NT-proBNP and either cTnT or cTnI (NT-proBNP < 332 ng/L, cTnT < 0.035 microg/L, and cTnI < 0.1 microg/L). Depending on whether NT proBNP and troponin levels were both low, were high for only one level, or were both high, patients were classified as stage I, II, or III, respectively. RESULTS: Using the cTnT+NT-proBNP model 33%, 30%, and 37% of patients were stages I, II, and III, respectively, with median survivals of 26.4, 10.5, and 3.5 months, respectively. The alternate cTnI+NT-proBNP model predicted median survivals of 27.2, 11.1, and 4.1 months, respectively. CONCLUSION: Stratification of AL patients into three stages is possible with two readily available and reproducible tests setting the stage for more consistent and reliable cross comparisons of therapeutic outcomes. PMID- 15365072 TI - Deleted in colon cancer protein expression in colorectal cancer metastases: a major predictor of survival in patients with unresectable metastatic disease receiving palliative fluorouracil-based chemotherapy. AB - PURPOSE: To determine whether deleted in colon cancer (DCC) protein expression in colorectal cancer (CRC) metastases could predict outcome to palliative fluorouracil (FU)-based chemotherapy and to assess whether it is similar to that observed in the corresponding primary tumors. PATIENTS AND METHODS: DCC protein expression was assessed immunohistochemically on archival specimens of CRC metastases from 42 patients homogeneously treated by methotrexate-modulated bolus FU alternated to 6-S-leucovorin-modulated infused FU and was retrospectively correlated with patient characteristics and clinical outcome. In a subset analysis, DCC immunoreactivity was compared between metastatic CRC and the corresponding primary tumors and regional lymph node metastases. RESULTS: Positive immunoreactivity for DCC was found in 45% of patients. Eighteen (78%) of 23 patients for whom multiple samples were available displayed a similar pattern of expression in distant metastases and primary tumors. The median survival time was 14.3 months in patients without DCC expression and 21.4 months in patients with DCC-positive tumors (log-rank test, P =.04); the 2-year survival rates were 8.5% and 42.5%, respectively. Response rates to chemotherapy were not significantly different between the two groups. By multivariate analysis, DCC protein expression maintained its prognostic value and showed to be the single best predictor of survival, with a relative risk of 2.16. CONCLUSION: Our results indicate that expression of the DCC protein in CRC metastases is similar to that observed in the corresponding primary tumors and represents a dominant predictor of survival in patients with unresectable, advanced CRC who are undergoing palliative FU-based chemotherapy. PMID- 15365073 TI - Modulation of fluorouracil by leucovorin in patients with advanced colorectal cancer: an updated meta-analysis. AB - PURPOSE: The modulation of fluorouracil (FU) by folinic acid (leucovorin [LV]) has been shown to be effective in terms of tumor response rate in patients with advanced colorectal cancer, but a meta-analysis of nine trials previously published by our group failed to demonstrate a statistically significant survival difference between FU and FU-LV. We present an update of the meta-analysis, with a longer follow-up and the inclusion of 10 newer trials. PATIENTS AND METHODS: Analyses are based on individual data from 3,300 patients randomized in 19 trials on an intent-to-treat basis. Two trials had multiple comparisons, leading to a total of 21 pair-wise comparisons. FU doses were similar in both arms in 10 pair wise comparisons, 15% to 33% higher in the FU-alone arm in six comparisons, and more than 66% higher in five comparisons. RESULTS: Overall analysis showed a two fold increase in tumor response rates (11% for FU-LV v 21% for FU-LV v 11% for FU [corrected] alone; odds ratio, 0.53; 95% CI, 0.44 to 0.63; P <.0001) and a small but statistically significant overall survival benefit for FU-LV over FU alone (median survival, 11.7 v 10.5 months, respectively; hazards ratio, 0.90; 95% CI, 0.87 to 0.94; P =.004), which were primarily seen in the first year. We observed a significant interaction between treatment benefit and dose of FU, with tumor response and overall survival advantages of FU-LV over FU-alone being restricted to trials in which a similar dose of FU was prescribed in both arms. CONCLUSION: This updated analysis demonstrates, on a large data set, that FU-LV improves both response rate and overall survival compared with FU alone and that this benefit is consistent across various prognostic factors. PMID- 15365074 TI - Irinotecan plus gemcitabine results in no survival advantage compared with gemcitabine monotherapy in patients with locally advanced or metastatic pancreatic cancer despite increased tumor response rate. AB - PURPOSE: This phase III, randomized, open-label, multicenter study compared the overall survival associated with irinotecan plus gemcitabine (IRINOGEM) versus gemcitabine monotherapy (GEM) in patients with chemotherapy-naive, locally advanced or metastatic pancreatic cancer. PATIENTS AND METHODS: IRINOGEM patients received starting doses of gemcitabine 1,000 mg/m2 and irinotecan 100 mg/m2 given weekly for 2 weeks every 3-week cycle. GEM patients received gemcitabine 1,000 mg/m2 weekly for 7 of 8 weeks (induction) and then weekly for 3 of 4 weeks. The primary end point of the trial was survival. Secondary end points included tumor response, time to tumor progression (TTP), changes in CA 19-9, and safety. RESULTS: In each arm, 180 randomly assigned patients comprised the intent-to treat population evaluated for efficacy; 173 IRINOGEM and 169 GEM patients were treated. Median survival times were 6.3 months for IRINOGEM (95% CI, 4.7 to 7.5 months) and 6.6 months for GEM (95% CI, 5.2 to 7.8 months; log-rank P =.789). Tumor response rates were 16.1% (95% CI, 11.1% to 22.3%) for IRINOGEM and 4.4% (95% CI, 1.9% to 8.6%) for GEM (chi2 P <.001). Median TTP was 3.5 months for IRINOGEM versus 3.0 months for GEM (log-rank P =.352). However, subset analyses in patients with locally advanced disease suggested a TTP advantage with IRINOGEM versus GEM (median, 7.7 v 3.9 months). CA 19-9 progression was positively correlated with tumor progression. The incidence of grade 3 diarrhea was higher in the IRINOGEM group but grade 3 to 4 hematologic toxicities and quality-of-life outcomes were similar. CONCLUSION: IRINOGEM safely improved the tumor response rate compared with GEM but did not alter overall survival. PMID- 15365075 TI - Early hospital discharge followed by outpatient management versus continued hospitalization of children with cancer, fever, and neutropenia at low risk for invasive bacterial infection. AB - PURPOSE: To compare outcome and cost of ambulatory versus hospitalized management among febrile neutropenic children at low risk for invasive bacterial infection (IBI). PATIENTS AND METHODS: Children presenting with febrile neutropenia at six hospitals in Santiago, Chile, were categorized as high or low risk for IBI. Low risk children were randomly assigned after 24 to 36 hours of hospitalization to receive ambulatory or hospitalized treatment and monitored until episode resolution. Outcome and cost were determined for each episode and compared between both groups using predefined definitions and questionnaires. RESULTS: A total of 161 (41%) of 390 febrile neutropenic episodes evaluated from June 2000 to February 2003 were classified as low risk, of which 149 were randomly assigned to ambulatory (n = 78) or hospital-based (n = 71) treatment. In both groups, mean age (ambulatory management, 55 months; hospital-based management, 66 months), sex, and type of cancer were similar. Outcome was favorable in 74 (95%) of 78 ambulatory-treated children and 67 (94%) of 71 hospital-treated children (P = NS). Mean cost of an episode was US 638 dollars (95% CI, 572 dollars to 703 dollars) and US 903 dollars (95% CI, 781 dollars to 1,025 dollars) for the ambulatory and hospital-based groups, respectively (P =.003). CONCLUSION: For children with febrile neutropenia at low risk for IBI, ambulatory management is safe and significantly cost saving compared with standard hospitalized therapy. PMID- 15365076 TI - Telomerase expression predicts unfavorable outcome in osteosarcoma. AB - PURPOSE: Osteosarcoma is distinct from most cancers in that the majority of osteosarcomas lack telomerase expression and use the alternative lengthening of telomeres (ALT) mechanism to maintain telomeres. Laboratory studies suggest that compared with ALT, telomerase expression is associated with increased tumor aggressiveness. We evaluated the clinical significance of telomerase expression in human osteosarcoma. PATIENTS AND METHODS: Fifty-six osteosarcomas from 51 patients treated at St Jude Children's Research Hospital between 1982 and 2003 were evaluated for telomerase enzyme activity, mRNA expression of the catalytic component of telomerase (TERT), and presence of the ALT pathway. RESULTS: Outcome analysis was based on TERT mRNA expression in the primary tumor samples from 44 patients. Fourteen primary tumors expressed TERT mRNA (32%; eight TERT only, six TERT and ALT) and 30 did not express TERT mRNA (68%; 29 ALT, one no ALT). Progression-free survival (PFS) was inferior in the TERT-positive group compared with the TERT-negative group (3-year estimates, 21.4% +/- 9.5% v 63.7% +/- 11.1%; P =.014). Likewise, overall survival was inferior in the TERT-positive group compared with the TERT-negative group (3-year estimates, 42.9% +/- 12.2% v 70.0% +/- 9.9%; P =.031). Among 31 patients with nonmetastatic disease at diagnosis, PFS was lower in the TERT-positive group compared with the TERT-negative group (3 year estimates, 33.3% +/- 13.6% v 72.0% +/- 11.5%; P =.092). CONCLUSION: Telomerase expression in primary tumor samples is associated with decreased PFS and OS in patients with osteosarcoma. Additional studies are warranted to better define the clinical utility of this molecular marker. PMID- 15365077 TI - Autologous hematopoietic stem-cell transplantation for children with acute myeloid leukemia in first or second complete remission: a prognostic factor analysis. AB - PURPOSE: To determine prognostic factors correlated with outcomes after autologous hematopoietic stem-cell transplantation (HSCT) in children with acute myeloid leukemia (AML). PATIENTS AND METHODS: We studied 219 children who received autologous HSCT for AML in first complete remission (CR) and 73 children in second CR and who were reported to the Autologous Blood and Marrow Transplant Registry. Among 29 of 73 patients who underwent transplantation in second CR, duration of first CR was > or = 12 months. RESULTS: Three-year cumulative incidences of relapse were 37% (95% CI, 31% to 44%), 60% (95% CI, 41% to 74%), and 36% (95% CI, 20% to 53%) for children in first CR, second CR after a short (< 12 months) first CR, and second CR after a long (> or = 12 months) first CR, respectively. Corresponding 3-year probabilities of leukemia-free survival were 54% (95% CI, 47% to 60%), 23% (95% CI, 10% to 39%), and 60% (95% CI, 42% to 75%). In multivariate analyses, risks of relapse, mortality, and treatment failure (relapse or death, inverse of leukemia-free survival) were higher for patients in second CR after a short first CR than for the other two groups. Transplant related mortality, treatment failure, and overall mortality rates were higher in older (> 10 years) children. CONCLUSION: Duration of first CR seems to be the most important determinant of outcome. Results in children who experience treatment failure with conventional chemotherapy support the use of autologous transplantation as salvage therapy if such patients achieve a subsequent CR. PMID- 15365078 TI - Systematic review of the staging performance of 18F-fluorodeoxyglucose positron emission tomography in esophageal cancer. AB - PURPOSE: Despite the increasing number of publications concerning (18)F fluorodeoxyglucose positron emission tomography (FDG-PET) for staging of esophageal cancer and the increasing availability of this novel diagnostic modality, its exact role in preoperative staging of these tumors is still unknown. The aim of this study was to systematically review the literature regarding the diagnostic performance of FDG-PET in preoperative staging of patients with esophageal cancer, and to calculate summary estimates of its sensitivity and specificity. METHODS: The databases of PubMed, Embase, and Cochrane were searched for relevant studies. Two reviewers independently assessed the methodological quality of each study. A meta-analysis of the reported sensitivity and specificity of each study was performed. RESULTS: Twelve studies met the inclusion criteria. The studies had several design deficiencies. Pooled sensitivity and specificity for the detection of locoregional metastases were 0.51 (95% CI, 0.34 to 0.69) and 0.84 (95% CI, 0.76 to 0.91), respectively. For distant metastases, pooled sensitivity and specificity were 0.67 (95% CI, 0.58 to 0.76) and 0.97 (95% CI, 0.90 to 1.0), respectively. CONCLUSION: FDG-PET showed moderate sensitivity and specificity for the detection of locoregional metastases, and reasonable sensitivity and specificity in detection of distant lymphatic and hematogenous metastases. PMID- 15365079 TI - Biology of gastrointestinal stromal tumors. AB - Once a poorly defined pathologic oddity, in recent years, gastrointestinal stromal tumor (GIST) has emerged as a distinct oncogenetic entity that is now center stage in clinical trials of kinase-targeted therapies. This review charts the rapid progress that has established GIST as a model for understanding the role of oncogenic kinase mutations in human tumorigenesis. Approximately 80% to 85% of GISTs harbor activating mutations of the KIT tyrosine kinase. In a series of 322 GISTs (including 140 previously published cases) studied by the authors in detail, mutations in the KIT gene occurred with decreasing frequency in exons 11 (66.1%), 9 (13%), 13 (1.2%), and 17 (0.6%). In the same series, a subset of tumors had mutations in the KIT-related kinase gene PDGF receptor alpha (PDGFRA), which occurred in either exon 18 (5.6%) or 12 (1.5%). The remainder of GISTs (12%) were wild type for both KIT and PDGFRA. Comparative studies of KIT-mutant, PDGFRA-mutant, and wild-type GISTs indicate that there are many similarities between these groups of tumors but also important differences. In particular, the responsiveness of GISTs to treatment with the kinase inhibitor imatinib varies substantially depending on the exonic location of the KIT or PDGFRA mutation. Given these differences, which have implications both for the diagnosis and treatment of GISTs, we propose a molecular-based classification of GIST. Recent studies of familial GIST, pediatric GIST, and variant forms of GIST related to Carney's triad and neurofibromatosis type 1 are discussed in relationship to this molecular classification. In addition, the role of mutation screening in KIT and PDGFRA as a diagnostic and prognostic aid is emphasized in this review. PMID- 15365080 TI - Challenging problems in malignancy: case 1. Presentation of small-cell lung cancer with marked hyperamylasemia. PMID- 15365081 TI - Challenging problems in malignancy: case 2. Isolated needle-track recurrence following fine needle aspiration for non-small-cell lung cancer. PMID- 15365082 TI - Challenging problems in malignancy: case 3. Chronic lymphocytic leukemia with ocular complications and superior vena cava obstruction. PMID- 15365083 TI - Exemestane following tamoxifen in postmenopausal women with primary breast cancer. PMID- 15365084 TI - Maturation matters: importance of maturation for antitumor immunity of dendritic cell vaccines. PMID- 15365086 TI - Risk of duodenal adenomas in familial adenomatous polyposis to progress toward advanced neoplastic disease. PMID- 15365088 TI - Cocaine induces apoptosis in fetal rat myocardial cells through the p38 mitogen activated protein kinase and mitochondrial/cytochrome c pathways. AB - Cocaine induces apoptosis in fetal rat myocardial cells (FRMCs). However, the mechanisms are not clear. The present study examined the role of p38 mitogen activated protein kinase (MAPK) and cytochrome c release in the cocaine-induced apoptosis in primary culture of FRMCs prepared from the fetal heart of gestational age of 21 days. Cocaine induced time-dependent, concurrent increases in cytochrome c release and activities of caspase-9 and caspase-3, which preceded apoptosis. Caspase-8 was not activated. In accordance, cyclosporin A and the inhibitors of caspase-9 and caspase-3 inhibited cocaine-induced caspase activation and apoptosis. Cocaine stimulated a transient increase in the p38 MAPK activity at a time point of 15 min but reduced the extracellular signal-regulated kinase (ERK) activity at 5 and 15 min in FRMCs. The p38alpha MAPK inhibitor SB203580 [4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H imidazole] inhibited cocaine-induced activation of caspases and apoptosis. In contrast, the p38beta MAPK and mitogen-activated protein kinase kinase/ERK inhibitors SB 202190 [4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)-1H imidazole] and PD98059 (2'-amino-3'-methoxyflavone), respectively, increased apoptosis in the absence of cocaine and potentiated cocaine-induced apoptosis. Consistent with its inhibition of apoptosis, SB203580 inhibited cocaine-induced cytochrome c release and activation of caspase-9 and caspase-3. In addition, cocaine induced a decrease in Bcl-2 protein levels, with no effect on Bax levels. The cocaine-mediated reduction of Bcl-2 levels was not affected with SB203580 and the caspase inhibitors. The results suggest that in FRMCs, p38alpha MAPK plays an important role in the cocaine-induced apoptosis by promoting cytochrome c release, downstream or independent of Bcl-2 protein-mediated regulation. In contrast, p38beta MAPK and ERK protect fetal myocardial cells against apoptosis. PMID- 15365089 TI - Human breast cancer resistance protein: interactions with steroid drugs, hormones, the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine, and transport of cimetidine. AB - The breast cancer resistance protein (BCRP/ABCG2) is an ATP-binding cassette drug efflux transporter that extrudes xenotoxins from cells, mediating drug resistance and affecting the pharmacological behavior of many compounds. To study the interaction of human wild-type BCRP with steroid drugs, hormones, and the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP), we expressed human BCRP in the murine MEF3.8 fibroblast cell line, which lacks Mdr1a/1b P glycoprotein and Mrp1, and in the polarized epithelial MDCKII cell line. We show that PhIP was efficiently transported by human BCRP in MDCKII-BCRP cells, as was found previously for murine Bcrp1. Furthermore, we show that six out of nine glucocorticoid drugs, corticosterone, and digoxin increased the accumulation of mitoxantrone in the MEF3.8-BCRP cell line, indicating inhibition of BCRP. In contrast, aldosterone and ursodeoxycholic acid had no significant effect on BCRP. The four most efficiently reversing glucocorticoid drugs (beclomethasone, 6alpha methylprednisolone, dexamethasone, and triamcinolone) and 17beta-estradiol showed a significantly reduced BCRP-mediated transepithelial transport of PhIP by MDCKII BCRP cells, with the highest reduction of PhIP transport ratio for beclomethasone (from 25.0 +/- 1.1 to 2.7 +/- 0.0). None of the tested endogenous steroids or synthetic glucocorticoids or digoxin, however, were transported substrates of BCRP. We also identified the H(2)-receptor antagonist drug cimetidine as a novel efficiently transported substrate for human BCRP and mouse Bcrp1. The generated BCRP-expressing cell lines thus provide valuable tools to study pharmacological and toxicological interactions mediated by BCRP and to identify new BCRP substrates. PMID- 15365090 TI - Inhibition of Na+-K+ pump and L-type Ca2+ channel by glibenclamide in Guinea pig ventricular myocytes. AB - Glibenclamide, a potent cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel blocker, is frequently used to study function and regulation of CFTR Cl(-) channels. In this study, the effects of glibenclamide on intracellular Na(+) concentration ([Na(+)](i)), contraction, Ca(2+) transient, and membrane potential were investigated in isolated guinea pig ventricular myocytes. Glibenclamide increased [Na(+)](i) and decreased contraction and Ca(2+) transient. However, glibenclamide did not change membrane potential. To determine whether inhibition of Na(+)-K(+) pumps and L-type Ca(2+) channels is responsible for the increase of [Na(+)](i) and the decrease of contraction, we tested the effects of glibenclamide on Na(+)-K(+) pump current and L-type Ca(2+) current (I(Ca,L)). Glibenclamide decreased Na(+)-K(+) pump current and I(Ca,L) in a concentration-dependent manner. In the presence of Cl(-) channel inhibitors, glibenclamide depolarized diastolic membrane potential and reduced action potential duration. This result suggests that the reason for lack of effect of glibenclamide on membrane potential might be due to its combined inhibitory effects on the Na(+)-K(+) pump, the L-type Ca(2+) channel, and Cl(-) channels, which may have opposing effects on membrane potential. These results indicate that glibenclamide increases [Na(+)(i)] by inhibiting the Na(+)-K(+) pump and decreases contraction and Ca(2+) transient, in addition, by blocking the L-type Ca(2+) channel. PMID- 15365091 TI - Sulodexide attenuates myocardial ischemia/reperfusion injury and the deposition of C-reactive protein in areas of infarction without affecting hemostasis. AB - Several glycosaminoglycans (GAGs) have been demonstrated to protect the ischemic heart against reperfusion injury, in part, by modulating activation of the complement cascade. The present study assessed the cardioprotective effects of sulodexide (KRX-101), a mixture of GAGs composed of 80% low-molecular mass heparin and 20% dermatan sulfate. KRX-101 differs from other GAGs (e.g., heparin) in that it has limited anticoagulant efficacy and can be administered orally. The experimental protocol was designed to determine whether KRX-101 could protect the ischemic myocardium. Anesthetized New Zealand white rabbits underwent 30 min of coronary artery occlusion. Intravenous doses of KRX-101 (0.5 mg/kg, n = 10) or drug diluent (n = 10) were administered at the end of regional ischemia and at each hour of reperfusion. Infarct size, as a percentage of the area at risk, was calculated for both groups. Myocardial infarct size was 31.3 +/- 4.1% in the vehicle- and 17.3 +/- 3.2% in the KRX-101-treated animals (p < 0.05 versus vehicle). Activated partial thromboplastin times determined at baseline (preischemia) and at each hour of reperfusion (n = 4) were not significantly different between vehicle- and KRX-101-treated groups (p = N.S.). Myocardial injury was further assessed by measuring serum levels of cardiac-specific troponin I. KRX-101 administration significantly reduced (p < 0.05) the serum concentration of troponin I during reperfusion. The results suggest that KRX-101 may be an effective adjunctive agent in myocardial revascularization procedures, without the risk of increased bleeding. PMID- 15365092 TI - Gating of the bacterial sodium channel, NaChBac: voltage-dependent charge movement and gating currents. AB - The bacterial sodium channel, NaChBac, from Bacillus halodurans provides an excellent model to study structure-function relationships of voltage-gated ion channels. It can be expressed in mammalian cells for functional studies as well as in bacterial cultures as starting material for protein purification for fine biochemical and biophysical studies. Macroscopic functional properties of NaChBac have been described previously (Ren, D., B. Navarro, H. Xu, L. Yue, Q. Shi, and D.E. Clapham. 2001. Science. 294:2372-2375). In this study, we report gating current properties of NaChBac expressed in COS-1 cells. Upon depolarization of the membrane, gating currents appeared as upward inflections preceding the ionic currents. Gating currents were detectable at -90 mV while holding at -150 mV. Charge-voltage (Q-V) curves showed sigmoidal dependence on voltage with gating charge saturating at -10 mV. Charge movement was shifted by -22 mV relative to the conductance-voltage curve, indicating the presence of more than one closed state. Consistent with this was the Cole-Moore shift of 533 micros observed for a change in preconditioning voltage from -160 to -80 mV. The total gating charge was estimated to be 16 elementary charges per channel. Charge immobilization caused by prolonged depolarization was also observed; Q-V curves were shifted by approximately -60 mV to hyperpolarized potentials when cells were held at 0 mV. The kinetic properties of NaChBac were simulated by simultaneous fit of sodium currents at various voltages to a sequential kinetic model. Gating current kinetics predicted from ionic current experiments resembled the experimental data, indicating that gating currents are coupled to activation of NaChBac and confirming the assertion that this channel undergoes several transitions between closed states before channel opening. The results indicate that NaChBac has several closed states with voltage-dependent transitions between them realized by translocation of gating charge that causes activation of the channel. PMID- 15365093 TI - Stabilizing the closed S6 gate in the Shaker Kv channel through modification of a hydrophobic seal. AB - The primary activation gate in K+ channels is thought to reside near the intracellular entrance to the ion conduction pore. In a previous study of the S6 activation gate in Shaker (Hackos et al., 2002), we found that mutation of V478 to W results in a channel that cannot conduct ions even though the voltage sensors are competent to translocate gating charge in response to membrane depolarization. In the present study we explore the mechanism underlying the nonconducting phenotype in V478W and compare it to that of W434F, a mutation located in an extracellular region of the pore that is nonconducting because the channel is predominantly found in an inactivated state. We began by examining whether the intracellular gate moves using probes that interact with the intracellular pore and by studying the inactivation properties of heterodimeric channels that are competent to conduct ions. The results of these experiments support distinct mechanisms underlying nonconduction in W434F and V478W, suggesting that the gate in V478W either remains closed, or that the mutation has created a large barrier to ion permeation in the open state. Single channel recordings for heterodimeric and double mutant constructs in which ion conduction is rescued suggest that the V478W mutation does not dramatically alter unitary conductance. Taken together, our results suggest that the V478W mutation causes a profound shift of the closed to open equilibrium toward the closed state. This mechanism is discussed in the context of the structure of this critical region in K+ channels. PMID- 15365094 TI - Mechanism of block of hEag1 K+ channels by imipramine and astemizole. AB - Ether a go-go (Eag; KV10.1) voltage-gated K+ channels have been detected in cancer cell lines of diverse origin and shown to influence their rate of proliferation. The tricyclic antidepressant imipramine and the antihistamine astemizole inhibit the current through Eag1 channels and reduce the proliferation of cancer cells. Here we describe the mechanism by which both drugs block human Eag1 (hEag1) channels. Even if both drugs differ in their affinity for hEag1 channels (IC50s are approximately 2 microM for imipramine and approximately 200 nM for astemizole) and in their blocking kinetics, both drugs permeate the membrane and inhibit the hEag1 current by selectively binding to open channels. Furthermore, both drugs are weak bases and the IC50s depend on both internal an external pH, suggesting that both substances cross the membrane in their uncharged form and act from inside the cell in their charged forms. Accordingly, the block by imipramine is voltage dependent and antagonized by intracellular TEA, consistent with imipramine binding in its charged form to a site located close to the inner end of the selectivity filter. Using inside- and outside-out patch recordings, we found that a permanently charged, quaternary derivative of imipramine (N-methyl-imipramine) only blocks channels from the intracellular side of the membrane. In contrast, the block by astemizole is voltage independent. However, as astemizole competes with imipramine and intracellular TEA for binding to the channel, it is proposed to interact with an overlapping intracellular binding site. The significance of these findings, in the context of structure function of channels of the eag family is discussed. PMID- 15365095 TI - Primary HIV-1 infection is associated with preferential depletion of CD4+ T lymphocytes from effector sites in the gastrointestinal tract. AB - Given its population of CCR5-expressing, immunologically activated CD4(+) T cells, the gastrointestinal (GI) mucosa is uniquely susceptible to human immunodeficiency virus (HIV)-1 infection. We undertook this study to assess whether a preferential depletion of mucosal CD4(+) T cells would be observed in HIV-1-infected subjects during the primary infection period, to examine the anatomic subcompartment from which these cells are depleted, and to examine whether suppressive highly active antiretroviral therapy could result in complete immune reconstitution in the mucosal compartment. Our results demonstrate that a significant and preferential depletion of mucosal CD4(+) T cells compared with peripheral blood CD4(+) T cells is seen during primary HIV-1 infection. CD4(+) T cell loss predominated in the effector subcompartment of the GI mucosa, in distinction to the inductive compartment, where HIV-1 RNA was present. Cross sectional analysis of a cohort of primary HIV-1 infection subjects showed that although chronic suppression of HIV-1 permits near-complete immune recovery of the peripheral blood CD4(+) T cell population, a significantly greater CD4(+) T cell loss remains in the GI mucosa, despite up to 5 yr of fully suppressive therapy. Given the importance of the mucosal compartment in HIV-1 pathogenesis, further study to elucidate the significance of the changes observed here is critical. PMID- 15365096 TI - CD4+ T cell depletion during all stages of HIV disease occurs predominantly in the gastrointestinal tract. AB - The mechanisms underlying CD4(+) T cell depletion in human immunodeficiency virus (HIV) infection are not well understood. Comparative studies of lymphoid tissues, where the vast majority of T cells reside, and peripheral blood can potentially illuminate the pathogenesis of HIV-associated disease. Here, we studied the effect of HIV infection on the activation and depletion of defined subsets of CD4(+) and CD8(+) T cells in the blood, gastrointestinal (GI) tract, and lymph node (LN). We also measured HIV-specific T cell frequencies in LNs and blood, and LN collagen deposition to define architectural changes associated with chronic inflammation. The major findings to emerge are the following: the GI tract has the most substantial CD4(+) T cell depletion at all stages of HIV disease; this depletion occurs preferentially within CCR5(+) CD4(+) T cells; HIV-associated immune activation results in abnormal accumulation of effector-type T cells within LNs; HIV-specific T cells in LNs do not account for all effector T cells; and T cell activation in LNs is associated with abnormal collagen deposition. Taken together, these findings define the nature and extent of CD4(+) T cell depletion in lymphoid tissue and point to mechanisms of profound depletion of specific T cell subsets related to elimination of CCR5(+) CD4(+) T cell targets and disruption of T cell homeostasis that accompanies chronic immune activation. PMID- 15365097 TI - The functional basis for hemophagocytic lymphohistiocytosis in a patient with co inherited missense mutations in the perforin (PFN1) gene. AB - About 30% of cases of the autosomal recessive immunodeficiency disorder hemophagocytic lymphohistiocytosis are believed to be caused by inactivating mutations of the perforin gene. We expressed perforin in rat basophil leukemia cells to define the basis of perforin dysfunction associated with two mutations, R225W and G429E, inherited by a compound heterozygote patient. Whereas RBL cells expressing wild-type perforin (67 kD) efficiently killed Jurkat target cells to which they were conjugated, the substitution to tryptophan at position 225 resulted in expression of a truncated ( approximately 45 kD) form of the protein, complete loss of cytotoxicity, and failure to traffic to rat basophil leukemia secretory granules. By contrast, G429E perforin was correctly processed, stored, and released, but the rat basophil leukemia cells possessed reduced cytotoxicity. The defective function of G429E perforin mapped downstream of exocytosis and was due to its reduced ability to bind lipid membranes in a calcium-dependent manner. This study elucidates the cellular basis for perforin dysfunctions in hemophagocytic lymphohistiocytosis and provides the means for studying structure function relationships for lymphocyte perforin. PMID- 15365098 TI - IL-7 receptor signals inhibit expression of transcription factors TCF-1, LEF-1, and RORgammat: impact on thymocyte development. AB - Intrathymic T cell development depends on signals transduced by both T cell receptor and cytokine receptors. Early CD4(-)CD8(-) (double negative) thymocytes require interleukin (IL)-7 receptor (IL-7R) signals for survival and proliferation, but IL-7R signals are normally extinguished by the immature single positive (ISP) stage of thymocyte development. We now demonstrate that IL-7R signals inhibit expression of transcription factors TCF-1, LEF-1, and RORgammat that are required for the ISP to double positive (DP) transition in the thymus. In addition, we demonstrate that IL-7R signals also inhibit TCF-1 and LEF-1 expression in mature peripheral T cells. Thus, the present work has identified several important downstream target genes of IL-7R signaling in T cells and thymocytes that provide a molecular mechanism for the inhibitory influence of IL 7R signaling on DP thymocyte development. We conclude that IL-7R signals down regulate transcription factors required for the ISP to DP transition and so must be terminated by the ISP stage of thymocyte development. PMID- 15365099 TI - Differential requirements for Vav proteins in DAP10- and ITAM-mediated NK cell cytotoxicity. AB - Natural killer (NK) cells express multiple activating receptors that initiate signaling cascades through DAP10- or immunoreceptor tyrosine-based activation motif-containing adapters, including DAP12 and FcRgamma. Among downstream signaling mediators, the guanine nucleotide exchange factor Vav1 carries out a key role in activation. However, whether Vav1 regulates only some or all NK cell activating pathways is matter of debate. It is also possible that two other Vav family molecules, Vav2 and Vav3, are involved in NK cell activation. Here, we examine the relative contribution of each of these exchange factors to NK cell mediated cytotoxicity using mice lacking one, two, or all three Vav proteins. We found that Vav1 deficiency is sufficient to disrupt DAP10-mediated cytotoxicity, whereas lack of Vav2 and Vav3 profoundly impairs FcRgamma- and DAP12-mediated cytotoxicity. Our results provide evidence that these three Vav proteins function specifically in distinct pathways that trigger NK cell cytotoxicity. PMID- 15365100 TI - Transitions and paradigms. PMID- 15365101 TI - Avian influenza: a human pandemic threat? PMID- 15365102 TI - Modulated release of health risk information to the general public with the use of mnemonics. PMID- 15365104 TI - Evidence for public health policy on inequalities: 1: the reality according to policymakers. AB - OBJECTIVE: To explore with UK and international policy advisors how research evidence influences public health policy making, and how its relevance and utility could be improved, with specific reference to the evidence on the production and reduction of health inequalities. DESIGN, SETTING, AND PARTICIPANTS: Qualitative residential workshop involving senior policy advisors with a substantive role in policy development across a range of sectors (mainly public health, but also including education, social welfare, and health services). In four in depth sessions, facilitated by the authors, focused questions were presented to participants. Their responses were then analysed thematically to identify key themes, relating to the availability and utility of existing evidence on health inequalities. MAIN RESULTS: The lack of an equity dimension in much aetiological and evaluative research was highlighted by participants. Much public health research was also felt to have weak underlying theoretical underpinnings. As well as evaluations of the effectiveness and cost effectiveness of policy and other interventions, they identified a need for predictive research, and for methodological research to further develop methods for assessing the impact on health of clusters of interventions. CONCLUSIONS: This study reinforces the view that there is a lack of information on the effectiveness and cost-effectiveness of policies, and it uncovered additional gaps in the health inequalities evidence base. A companion paper discusses researchers' views of how the production of more relevant public health evidence can be stimulated. PMID- 15365105 TI - Evidence for public health policy on inequalities: 2: assembling the evidence jigsaw. AB - STUDY OBJECTIVE: To garner research leaders' perceptions and experiences of the types of evidence that influence policy on health inequalities, and their reflections on how the flow of such research evidence could be increased. DESIGN, SETTING, AND PARTICIPANTS: Qualitative two day residential workshop with senior research leaders, most of whom were currently involved in evaluations of the health effects of major policies. In four in depth sessions, facilitated by the authors in turn, focused questions were presented to participants to reveal their views and experiences concerning evidence synthesis for policy on inequalities. These were analysed thematically. MAIN RESULTS: Five types of evidence for policy on health inequalities were felt to be particularly persuasive with policymakers: observational evidence showing the existing of a problem; narrative accounts of the impacts of policies from the household perspective; controlled evaluations; natural policy experiments; and historical evidence. Methods of improving the availability and use of these sources of information were put forward. CONCLUSIONS: This paper and its companion have considered the current evidence base for policies to reduce health inequalities, and how this could be improved. There is striking congruence between the views of the researchers in this study and policy advisers in paper 1, suggesting that a common understanding may be emerging. The findings suggest significant potential for rapid progress to be made in developing both evidence based policy, and policy relevant evidence to tackle inequalities in health. PMID- 15365106 TI - To control and beyond: moving towards eliminating the global tuberculosis threat. AB - For 10 years the World Health Organisation has had a single answer to the deadly threat of tuberculosis (TB)-provide treatment to smear positive patients and watch them take it. In contrast with confident statements about how global TB would be brought under control when directly observed therapy, short course (DOTS) was introduced, TB continues to rise worldwide. The introduction of selected multiple drug resistant TB treatment programmes, "DOTS-Plus", although important, also focuses on therapy for active TB. HIV endemic countries in particular have experienced tremendous increases in TB despite having DOTS programmes. A critical review of recent epidemiological data and computer models shows that the present international strategy of concentrating on providing treatment for smear positive TB, DOTS and DOTS-Plus, is likely to have only a modest impact on population based TB control. Effective global TB control will require strategies that go beyond relying on treatment of people with active disease. PMID- 15365107 TI - Malaria, from natural to supernatural: a qualitative study of mothers' reactions to fever (Dienga, Gabon). AB - OBJECTIVE: Decision making for health care at the household level is a crucial factor for malaria management and control among young children. This study sought to determine exactly how mothers reacted when faced with fever in a child. DESIGN: Qualitative study based on in depth semistructured interviews of mothers and free form discussion with traditional healers (Nganga). SETTING: Village of Dienga, a rural area of Gabon (Central Africa). PARTICIPANTS: 12 mothers and three traditional healers. RESULTS: All mothers thought that fever and malaria were identical. Mothers home treated or went to the village treatment centre, or both, on the last episode of fever, if they judged it to be "natural" fever. However, if fever was thought to be a result of malicious intent, then a Nganga was consulted first. It was believed that strong and above all persistent fever was "supernatural". In this case, traditional treatment was thought to be best. CONCLUSIONS: Results indicate that fever is perceived as a dual condition, with two distinct but non-mutually exclusive aetiologies (either "natural" or from witchcraft). In contrast with what is commonly believed, there seems to be no clear cut distinction between diseases suitable for management by western medicine and diseases to be managed solely by traditional health practitioners. Moreover, these data do not support the commonly held notion that the decision to seek western medicine to treat fever is considered a "last resort". Results strongly imply that some severe cases of fever, being initially considered supernatural, may partially or completely escape medical attention. PMID- 15365108 TI - Apheis: public health impact of PM10 in 19 European cities. AB - STUDY OBJECTIVE: Apheis is a public health surveillance system that aims to provide European, national, regional, and local decision makers, environmental health professionals, and the general public with up to date and easy to use information on air pollution and public health. This study presents the health impact assessment done in 19 cities of Western and Eastern European countries. DESIGN: Apheis developed guidelines for gathering and analysing data on air pollution and the impact on public health. Apheis has analysed the acute and chronic effects of fine particles on premature mortality using the estimates developed by Aphea2 study and two American cohort studies. This health impact assessment was performed for different scenarios on the health benefits of reducing levels of particles less than 10 microm in size (PM(10)). MAIN RESULTS: PM(10) concentrations were measured in 19 cities (range: 14-73 microg/m(3)). The population covered in this health impact assessment includes nearly 32 million inhabitants. The age standardised mortality rates (per 100 000 people) range from 456 in Toulouse to 1127 in Bucharest. Reducing long term exposure to PM(10) concentrations by 5 microg/m(3) would have "prevented" between 3300 and 7700 early deaths annually, 500 to 1000 of which are associated with short term exposure. CONCLUSIONS: Apheis shows that current levels of air pollution in urban Europe have a non-negligible impact on public health, and that preventive measures could reduce this impact, even in cities with low levels of air pollution. PMID- 15365109 TI - Evaluation of the health effects of a neighbourhood traffic calming scheme. AB - STUDY OBJECTIVE: To assess the secondary health impacts of a traffic calming scheme on a community. METHODS: Prospective cohort study of a randomly selected sample of the local community using postal questionnaires and pedestrian counts on the affected road six months before and six months after the implementation of the scheme. The setting was a community in which a traffic calming scheme was built in the main road (2587 households). The Short Form 36 version 2 was included in the questionnaire and summary measures of physical health (physical component summary) and mental health (mental component summary) calculated. A random sample of 750 households was initially posted the pre-intervention questionnaire. MAIN RESULTS: There were increases in observed pedestrian activity in the area after the introduction of the traffic calming scheme. Physical health improved significantly but mental health did not change. Traffic related problems improved, while other local nuisances were reported to be worse. CONCLUSIONS: The introduction of a traffic calming scheme is associated with improvements in health and health related behaviours. It is feasible to prospectively evaluate broader health impacts of similar transport interventions although poor response rates may limit the validity of results. PMID- 15365111 TI - Post-intervention effect of a computer tailored smoking cessation programme. PMID- 15365110 TI - Safer storage of firearms at home and risk of suicide: a study of protective factors in a nationally representative sample. AB - OBJECTIVE: To estimate the protective effect of storing firearms locked or unloaded, or both, on the risk of suicide by firearms among people with relatively low intention to die. DESIGN AND SETTING: Cross sectional survey. The 1993 National Mortality Followback Survey of 22 957 deaths in the United States, representing 2.2 million people, conducted by the National Center for Health Statistics. PARTICIPANTS: Decedent's next of kin answered questions regarding various aspects of decedent's life to supplement information from death certificates. MAIN RESULTS: Compared with decedents who stored their firearm unlocked or loaded, those who stored their firearms locked or unloaded, or both, were less likely to commit suicide by firearms (locked: OR = 0.39, 95% CI = 0.24 to 0.66; unloaded OR = 0.30, 95% CI = 0.18 to 0.49). CONCLUSIONS: This study further supports the utility of devices and practices intended to reduce the likelihood of unauthorised or impulsive use of firearms. PMID- 15365112 TI - Infections, medication use, and the prevalence of symptoms of asthma, rhinitis, and eczema in childhood. AB - BACKGROUND: The "hygiene hypothesis" postulates that infections during infancy may protect against asthma and atopy. There is also some evidence that antibiotic and/or paracetamol use may increase the risk of asthma. METHODS: The study measured the association between infections, and medication use early in life and the risk of asthma at age 6-7 years. It involved 1584 children who had been notified to public health services with serious infections at age 0-4 years, and 2539 children sampled from the general population. For both groups, postal questionnaires were completed by parents. RESULTS: There was little difference in the prevalence of current wheezing between the childhood infections group (prevalence = 23.5%) and the general population group (prevalence = 24.3%). There was also little difference whether the major site of infection was gastrointestinal (prevalence = 24.1%), invasive (prevalence = 24.6%) or respiratory (prevalence = 21.1%). However, in both groups, there were associations with antibiotic (OR = 1.78, 95% CI 1.49 to 2.14) or paracetamol (OR = 1.38, 95% CI 1.04 to 1.83) use in the first year of life or recent paracetamol use (OR = 2.10, 95% CI 1.78 to 2.49) and current wheezing. There was a weak protective effect of childhood infections in children who had not used antibiotics in the first year of life (OR = 0.78, 95% CI 0.55 to 1.10). CONCLUSIONS: These findings are consistent with other evidence that antibiotic use early in life may increase the risk of asthma. They are also consistent with some preliminary evidence associating paracetamol use with an increased risk of asthma. Any protective effect of notifiable childhood infections was weak. PMID- 15365113 TI - Measures of health inequalities: part 1. PMID- 15365114 TI - Socioeconomic inequalities in mobility decline in chronic disease groups (asthma/COPD, heart disease, diabetes mellitus, low back pain): only a minor role for disease severity and comorbidity. AB - OBJECTIVE: This study examined the association between socioeconomic status and mobility decline and whether this could be explained by disease severity and comorbidity in four different chronic disease groups (asthma/COPD, heart disease, diabetes mellitus, and low back pain). It is not clear, whether the adverse course of physical functioning in persons with a low socioeconomic status can be explained by a higher prevalence of more severe disease or comorbidity in these persons. DESIGN: Dutch GLOBE study: prospective cohort study. SETTING: Region of Eindhoven (south east of the Netherlands). PARTICIPANTS: 1384 persons suffering from at least one of the four chronic diseases were selected. The number of respondents in each group was: asthma/COPD 465, heart disease 788, diabetes mellitus 137, and low back pain 707. There were 580 respondents who suffered from more than one condition. MAIN RESULTS: Odds ratios of mobility decline between 1991 and 1997, adjusted for age, sex, marital status, and baseline mobility, were significantly higher in low socioeconomic groups in comparison with high socioeconomic groups. Only very little of this association could be explained by the higher disease severity and comorbidity in these patients. Findings were similar in patients with asthma/COPD, heart disease, diabetes mellitus, and chronic low back pain. CONCLUSION: These findings indicate that to reduce physical disabilities and particularly the socioeconomic differences therein, it may not be sufficient to solely intervene upon the risks of severe disease and comorbidities. PMID- 15365115 TI - Psychosocial factors and work related sickness absence among permanent and non permanent employees. AB - STUDY OBJECTIVE: To examine the association between psychosocial work factors and work related sickness absence among permanent and non-permanent employees by sex. DESIGN: A cross sectional survey conducted in 2000 of a representative sample of the European Union total active population, aged 15 years and older. The independent variables were psychological job demands and job control as measures of psychosocial work environment, and work related sickness absence as the main outcome. Poisson regression models were used to compute sickness absence days' rate ratios. SETTING: 15 countries of the European Union. PARTICIPANTS: A sample of permanent (n = 12 875) and non-permanent (n = 1203) workers from the Third European Survey on Working Conditions. RESULTS: High psychological job demands, low job control, and high strain and passive work were associated with higher work related sickness absence. The risks were more pronounced in non-permanent compared with permanent employees and men compared with women. CONCLUSIONS: This work extends previous research on employment contracts and sickness absence, suggesting different effects depending on psychosocial working conditions and sex. PMID- 15365116 TI - Development of the chronic fatigue syndrome in severely fatigued employees: predictors of outcome in the Maastricht cohort study. AB - STUDY OBJECTIVE: To identify risk factors of the development of the chronic fatigue syndrome (CFS), the persistence or recurrence of fatigue, or recovery from fatigue in a large sample of fatigued employees. DESIGN: Analyses were based on the Maastricht cohort study (MCS), a prospective population based cohort study among more than 12 000 employees. Multiple regression models were used to identify predictors of CFS-like caseness (meeting research criteria for CFS), non CFS fatigue caseness, or no fatigue caseness. SETTING: The working population in the Netherlands. PARTICIPANTS: 1143 employees with medically unexplained fatigue were followed up prospectively for 44 months. MAIN RESULTS: At 44 month follow up, 8% of the employees were CFS-like cases (none of who reported to have received a CFS diagnosis), 40% were non-CFS fatigue cases, and 52% were no longer fatigue cases. Factors that predicted CFS-like caseness compared with non-CFS fatigue caseness were high age, exhaustion, female sex, low education, and visits to the general practitioner. Factors that predicted CFS-like caseness compared with no fatigue caseness were fatigue, exhaustion, low education, visits to the GP and occupational physician, and bad self rated health. Factors that predicted non-CFS fatigue caseness compared with no fatigue caseness were fatigue, low self perceived activity, exhaustion, anxious mood, and bad self rated health. CONCLUSIONS: Unexplained fatigue among employees in some instances is a precursor of the development of CFS. The prognostic role of self rated health suggests that prevention and treatment of chronic fatigue should be aimed at changing the perception of health or illness. Less clear is the role of health care seeking or receiving a CFS diagnosis. PMID- 15365117 TI - Fibrinogen, social position, and "Mendelian randomisation". PMID- 15365119 TI - Thrombin: maybe not so spellbinding. PMID- 15365120 TI - How not to synthesize synuclein. PMID- 15365121 TI - Does the hepatitis B vaccine cause multiple sclerosis? PMID- 15365122 TI - Vascular neurology: a new neurologic subspecialty. AB - This article presents information about the American Board of Psychiatry and Neurology's new subspecialty certificate in vascular neurology, including the requirements for board certification. Practicing neurologists and child neurologists who have a major focus in vascular neurology can apply for the new certificate. In addition, the Accreditation Council for Graduate Medical Education has approved fellowship training in the subspecialty, and it is likely that residents who wish to pursue a career in vascular neurology will select this training option. PMID- 15365123 TI - Thrombin generation in non-cardioembolic stroke subtypes: the Hemostatic System Activation Study. AB - BACKGROUND: The association between hemostatic activation, stroke mechanism, and outcome is poorly defined. The Hemostatic System Activation Study (HAS) investigators measured serial levels of prothrombin fragment F1.2, a marker of thrombin generation, in patients enrolled in the Warfarin Aspirin Recurrent Stroke Study (WARSS). METHODS: HAS enrolled 631 of the 2,206 patients in WARSS. Strokes were subtyped according to inferred mechanism. Plasma was collected for F1.2 at randomization (within 30 days of stroke), 3 months, 12 months, and 18 months. The 3 to 6 month samples in aspirin-treated patients were used for the primary analysis. RESULTS: The authors analyzed 3 to 6 month samples on 320 patients. Higher F1.2 levels were associated with older age, female sex, and hypertension. There was no difference between mean F1.2 levels in 56 cryptogenic (0.9 +/- 0.32 nmol/L) and 114 non-cryptogenic (1.13 +/- 0.74 nmol/L) patients or across specific stroke subtypes. There was an 8.8%/year (p = 0.006) increase in mean F1.2 levels. There was a trend toward higher risk of recurrent stroke or death as F1.2 levels increased in aspirin (RR: 1.30, 95% CI: 0.57 to 2.94, p = 0.53) and warfarin treated patients (RR: 1.68, 95% CI: 0.48 to 5.94, p = 0.42). F1.2 levels were reduced on average 70% in warfarin-treated patients in a dose dependent fashion. CONCLUSION: F1.2 levels did not appear to differ by stroke subtype, suggesting that factors other than underlying stroke pathophysiology influence thrombin generation in the post-acute stroke period. F1.2 levels were suppressed by warfarin in a dose-dependent fashion. Additional research is needed to determine the predictive value of F1.2 after stroke. PMID- 15365124 TI - Progressive dementia after first-ever stroke: a community-based follow-up study. AB - OBJECTIVES: To examine the risk and determinants of a progressive dementia syndrome and cognitive impairment not dementia (CIND) in community-based nonaphasic first-ever stroke cases 1 year after stroke, relative to a matched community-based stroke-free group. METHODS: Matched cohort design, with cognitive tests given on two occasions 9 months apart to 99 mild-to-moderate first-ever stroke patients and 99 age- and sex-matched people without stroke. At follow-up, progressive dementia or CIND were diagnosed, with judges blinded to stroke/nonstroke status. RESULTS: Progressive dementia was diagnosed in 12.5% of stroke patients and 15.4% of those without strokes (RR 1.1, 95% CI 0.5 to 2.2, p = 0.85). CIND was diagnosed in 37.5% of stroke patients and 17.6% of participants without strokes (RR 2.1, 95% CI 1.2 to 3.4, p = 0.003). In multivariable regression, age (p = 0.04) and baseline cognition (p < 0.001) were independently associated with dementia whereas stroke (p = 0.002), age (p = 0.05), baseline cognition (p = 0.001), and baseline mood (p = 0.03) were independently associated with CIND at follow-up. CONCLUSIONS: In this community-based nonaphasic sample, mild-to moderate first-ever stroke was not associated with the presence of progressive dementia 1 year later, but was clearly associated with a greater risk of cognitive impairment not dementia (CIND). Baseline mood impairment remained independently associated with CIND at 1 year after taking into account stroke, age, and baseline cognitive ability. PMID- 15365125 TI - Stroke risk factors and loss of high cognitive function. AB - BACKGROUND: Modifiable stroke risk factors may contribute to age-associated declines in cognitive function. Individuals with high levels of cognitive function after midlife may have less exposure to these stroke risk factors or may be less susceptible to their effects on cognition. METHODS: The Cardiovascular Health Study (CHS)* is a population-based, longitudinal cohort study of 5,888 people age 65 years and older. Participants (n = 4,129) who were free of dementia, stroke, or TIA at the time of baseline cranial MRI were selected for analysis. High cognitive function at baseline was defined by performance at or above midlife norms on the Modified Mini-Mental State Examination (3MS). RESULTS: The odds of having high cognitive function at baseline decreased by quartile of stroke risk (highest vs lowest risk quartile, adjusted odds ratio [OR] 0.68; 95% CI 0.52 to 0.88; p for trend = 0.005). Stroke risk was a predictor of decline on the 3MS in those with typical levels of cognitive function at baseline, even in the absence of incident stroke or TIA (highest vs lowest risk quartile for 3MS decline, adjusted OR 2.11; 95% CI 1.42 to 3.13; p for trend < 0.001). In contrast, stroke risk was not associated with decline on the 3MS in those with high cognitive function at baseline (p = 0.03 for interaction). CONCLUSIONS: In a cohort of older adults without stroke, TIA, or dementia, cognitive function and incident cognitive decline were associated with risk for stroke. Additional studies are needed to determine whether modification of stroke risk factors can reduce the cognitive decline that is often attributed to normal aging. PMID- 15365126 TI - CagA-positive Helicobacter pylori strains may influence the natural history of atherosclerotic stroke. AB - OBJECTIVE: To test the hypothesis that infection with virulent cytotoxin associated gene-A (CagA)-bearing Helicobacter pylori strains influences the atherosclerotic process and the clinical course in atherosclerotic stroke patients. METHODS: ELISA was used to assess the seroprevalence of infection by H. pylori and CagA-positive strains in 185 patients. Intima-media thickness (IMT) was determined by Doppler ultrasound. Baseline, 1-week, and 1-month NIH Stroke Scale (NIHSS) scores were used to evaluate the short-term clinical course. RESULTS: H. pylori infection was found in 79% of patients; 58% of these tested positive for CagA. IMT was higher among CagA-positive patients than among CagA negative ones (1.13 +/- 0.26 mm vs 0.97 +/- 0.15 mm; univariate analysis, p = 0.0001; multivariate analysis, odds ratio [OR], 2.36; 95% CI, 1.57 to 3.54; p = 0.0001) or H. pylori-negative ones (1.01 +/- 0.17 mm; univariate analysis, p = 0.007; multivariate analysis, OR, 1.90; 95% CI, 1.22 to 2.97; p = 0.005). CagA positive patients had poorer initial outcomes based on serial measurements of the NIHSS score (repeated measures analysis of variance, p < 0.0001). No significant difference in IMT and NIHSS score was found between H. pylori-positive and H. pylori-negative patients. CONCLUSIONS: Infection with cytotoxin-associated gene-A positive Helicobacter pylori strains in atherosclerotic stroke patients is associated with greater intima-media thickness and poorer short-term outcome compared with cytotoxin-associated gene-A-negative patients. PMID- 15365127 TI - Beta-synuclein gene alterations in dementia with Lewy bodies. AB - OBJECTIVE: To determine whether mutations in the genes for alpha-synuclein or beta-synuclein are responsible for dementia with Lewy bodies (DLB), a disorder closely related to Parkinson disease (PD). METHODS: The authors ascertained 33 sporadic cases of DLB and 10 kindreds segregating DLB. DNA samples from the 43 index cases were screened for alterations in the genes for alpha-synuclein and beta-synuclein, as alpha-synuclein alterations cause PD and beta-synuclein may modulate alpha-synuclein aggregation and neurotoxicity. RESULTS: Two amino acid alterations were identified in unrelated DLB index cases: a valine to methionine substitution at codon 70 (V70M) and a proline to histidine substitution at codon 123 (P123H), both in the beta-synuclein gene. These amino acid substitutions occur at conserved residues in highly conserved regions of the beta-synuclein protein. Screening of at least 660 chromosomes from control subjects matched to the patients' population groups failed to identify another V70M or P123H allele. Cosegregation analysis of an extended pedigree segregating the P123H beta synuclein alteration suggested that it is a dominant trait with reduced penetrance or a risk factor polymorphism. Histopathology and immunohistochemistry analysis of index case brain sections revealed widespread Lewy body pathology and alpha-synuclein aggregation without evidence of beta-synuclein aggregation. CONCLUSION: Mutations in the beta-synuclein gene may predispose to DLB. PMID- 15365128 TI - Two different pathogenetic mechanisms in psychogenic tremor. AB - BACKGROUND: The pathophysiologic mechanism underlying psychogenic tremor is not clear. Continuous voluntary production of tremor may be uncovered by a positive entrainment of tremor in different limbs. But some patients have tremor ongoing during their waking time which is unlikely to be produced voluntarily. Therefore, nonvoluntary physiologic oscillatory mechanisms must be considered. METHODS: Fifteen patients with psychogenic tremor manifesting in both hands, who were diagnosed using established criteria, were examined. Postural tremor was recorded with accelerometry and electromyography (EMG) while the hands were held against gravity. Power spectral peak frequencies and accelerometric total power as a measure of amplitude were determined. Coherency spectra between the EMG signals from the right and left arm were calculated. RESULTS: Seven of 15 patients showed a significant coherency between the two hands; the remaining 8 patients maintained independent oscillations. Clinical presentation, tremor frequencies, and amplitudes were not significantly different between the two groups. CONCLUSIONS: Two different pathogenetic mechanisms may play a role in psychogenic tremor. Bilateral voluntary movements are typically coherent. Thus, coherent psychogenic tremor would be in keeping with voluntarily produced oscillations. Absent coherence is an indication of another, possibly nonvoluntary mechanism like clonus or enhanced physiologic tremor. PMID- 15365129 TI - APOE and cognitive decline in preclinical Alzheimer disease and non-demented aging. AB - OBJECTIVE: To investigate whether presence of the APOE epsilon4 allele is related to the pathologic progression of preclinical Alzheimer disease (AD), as reflected by change in Mini-Mental State Examination (MMSE) scores among persons in the preclinical phase of AD, and cognitively intact adults confirmed as dementia-free during the 6-year assessment period. METHOD: In a population-based sample, participants were stratified according to APOE genotype (epsilon4 or non epsilon4) and whether they received a diagnosis of AD at the end of either a 3- or 6-year assessment period. Participants were aged 75 years and older, and were nondemented at baseline. At the end of the 3-year period, 17.2% of non-epsilon4 and 26.7% of epsilon4 carriers became demented. For the 6-year period those percentages were 11.2% for non-epsilon4 carriers and 16.9% for epsilon4-carriers. RESULTS: Individuals in the preclinical phase of AD showed greater decline on the MMSE as compared to nondemented adults. However, the decline was most marked in the 3 years prior to clinical diagnosis. Further, APOE-epsilon4 genotype did not modify the rate of decline among to-be-demented participants, as well as individuals who would remain free of AD. CONCLUSIONS: Although possession of the APOE epsilon4 allele is a risk factor for AD in old age, it does not modify the progression of the disease during the preclinical period. Further, in the absence of preclinical dementia, APOE did not influence global cognitive change in nondemented persons. PMID- 15365130 TI - Higher frequency of dementia in older HIV-1 individuals: the Hawaii Aging with HIV-1 Cohort. AB - BACKGROUND: Antiretroviral therapy has improved survival for HIV-1-infected individuals. The neuroepidemiologic implications of HIV-1 in an aging population are not well known, particularly the prevalence of HIV-associated dementia (HAD). METHODS: The authors report a baseline cross-sectional analysis of 202 HIV-1 seropositive individuals enrolled into one of two groups of the Hawaii Aging with HIV Cohort: older (50 or more years old, n = 106) and younger (20 to 39 years old, n = 96). Neuropsychological, neurologic, medical, and laboratory data were obtained at enrollment. Participant cognitive status was classified (research case definitions) using American Academy of Neurology (1991) criteria in a consensus conference of physicians and neuropsychologists. RESULTS: HAD was more frequent in older (25.2%) compared to younger (13.7%) individuals (p = 0.041) corresponding to an OR of 2.13 (95% CI: 1.02 to 4.44) for the older compared to the younger group. After adjusting for education, race, substance dependence, antiretroviral medication status, viral load, CD4 lymphocyte count, and Beck Depression Inventory score, the odds of having HAD among individuals in the older group was 3.26 (1.32 to 8.07) times that of the younger group. CONCLUSIONS: Older age is associated with increased HAD in this HIV-1 cohort. Underlying mechanisms are unclear but do not appear related to duration of HIV-1 infection. PMID- 15365131 TI - Plasma levels of amyloid beta-protein 42 are increased in women with mild cognitive impairment. AB - BACKGROUND: Accumulation in the brain of small aggregates of amyloid beta-protein 42 (Abeta42) is the major pathogenic event of Alzheimer disease (AD). In familial early-onset AD this event is likely the result of Abeta42 overproduction; in the most common sporadic late-onset form of the disease the mechanisms of Abeta42 accumulation are unknown. METHODS: To address this issue the authors analyzed plasma levels of Abeta42 in 88 elderly patients with amnestic mild cognitive impairment (MCI), chosen as paradigm of preclinical sporadic AD. RESULTS: The authors found a significant increase of Abeta42 plasma levels in women with MCI, in comparison to the affected men and 72 cognitively normal age-matched subjects. The levels were independent of variables in education, apolipoprotein E genotype, cholesterol, and creatinine plasma concentrations, as well as hemoglobin content. CONCLUSIONS: The elevation of Abeta42 plasma levels in women with MCI may represent a biologic explanation for the sex-dependent increased incidence of late-onset AD in women identified by epidemiologic studies. PMID- 15365132 TI - Driver route-following and safety errors in early Alzheimer disease. AB - OBJECTIVE: To assess navigation and safety errors during a route-following task in drivers with Alzheimer disease (AD). DESIGN/METHODS: Thirty-two subjects with probable AD (by National Institute of Neurological and Communicative Disorders criteria) of mild severity and 136 neurologically normal older adults were tested on a battery of visual and cognitive tests of abilities that are critical to safe automobile driving. Each driver also performed a route-finding task administered on the road in an instrumented vehicle. Main outcome variables were number of 1) incorrect turns; 2) times lost; and 3) at-fault safety errors. RESULTS: The drivers with mild AD made significantly more incorrect turns, got lost more often, and made more at-fault safety errors than control subjects, although their basic vehicular control abilities were normal. The navigational and safety errors were predicted using scores on standardized tests sensitive to visual and cognitive decline in early AD. CONCLUSIONS: Drivers with Alzheimer disease made more errors than neurologically normal drivers on a route-following task that placed demands on driver memory, attention, and perception. The demands of following route directions probably increased the cognitive load during driving, which might explain the higher number of safety errors. PMID- 15365133 TI - Recombinant hepatitis B vaccine and the risk of multiple sclerosis: a prospective study. AB - BACKGROUND: A potential link between the recombinant hepatitis B vaccine and an increased risk of multiple sclerosis (MS) has been evaluated in several studies, but some of them have substantial methodologic limitations. METHODS: The authors conducted a nested case-control study within the General Practice Research Database (GPRD) in the United Kingdom. The authors identified patients who had a first MS diagnosis recorded in the GPRD between January 1993 and December 2000. Cases were patients with a diagnosis of MS confirmed through examination of medical records, and with at least 3 years of continuous recording in the GPRD before their date of first symptoms (index date). Up to 10 controls per case were randomly selected, matched on age, sex, practice, and date of joining the practice. Information on receipt of immunizations was obtained from the computer records. RESULTS: The analyses include 163 cases of MS and 1,604 controls. The OR of MS for vaccination within 3 years before the index date compared to no vaccination was 3.1 (95% CI 1.5, 6.3). No increased risk of MS was associated with tetanus and influenza vaccinations. CONCLUSIONS: These findings are consistent with the hypothesis that immunization with the recombinant hepatitis B vaccine is associated with an increased risk of MS, and challenge the idea that the relation between hepatitis B vaccination and risk of MS is well understood. PMID- 15365134 TI - Primary chronic daily headache and its subtypes in adolescents and adults. AB - OBJECTIVES: To determine the relative frequency of chronic daily headache (CDH) subtypes in adolescents and to compare the distribution of CDH subtypes in adolescents and adults of various ages. METHODS: Adolescents (13 to 17 years, n = 170) and adults (18 or older, n = 638) were recruited during the same time frame. CDH subtypes were classified according the criteria proposed by Silberstein and Lipton (1996) as transformed migraine (TM), chronic tension-type headache (CTTH), new daily persistent headache (NDPH), and hemicrania continua (HC). RESULTS: Among adolescents and adults there were substantial differences in the distribution of CDH subtypes. The relative frequency of TM was lower in adolescents (68.8% vs 87.4%, p < 0.001), while NDPH (21.1% vs 10.8%, p < 0.001) and CTTH (10.1% vs 0.9%, p < 0.0001) were more common. HC (0 vs 0.9%, NS) was equally rare. The lower relative frequency of TM in adolescents was accounted for by TM with medication overuse (TM+), much more common in adults (28.2% vs 62.5%, p < 0.001). In fact, TM without medication overuse (TM-) was more common in adolescents (40.5% vs 24.9%, p < 0.001). The relative frequency of TM+ increased until the age of 50 years (p < 0.001). CONCLUSIONS: In adolescents with CDH, TM usually develops without medication overuse. Adolescents with the early onset form of TM may develop the disorder in the absence of medication overuse because they are at increased biologic risk. PMID- 15365135 TI - Clinical recognition of allodynia in migraine. AB - BACKGROUND: Allodynia, a manifestation of central sensitization, is not routinely evaluated during clinical interviews even though its therapeutic implications are known. OBJECTIVE: To assess the frequency and manifestations of allodynia in a clinic-based migraine population. METHODS: A total of 295 consecutive International Headache Society migraine patients were evaluated using a semistructured questionnaire. RESULTS: A total of 157 (53.3%) reported allodynia. A total of 133 (84.7% of those with allodynia) had cephalic, 54 (18%) had both cephalic and extracephalic, and 24 (8.5%) exhibited pure extracephalic allodynia. Prominent scalp symptoms were sensitivity to touch; soreness or tenderness; difficulty brushing, combing, or washing hair; difficulty lying with head resting on the side of allodynia; and circumscribed area of hypersensitivity and tenderness. Scalp allodynia was ipsilateral to predominant headache side in the majority and occurred at the height of headache. Extracephalic allodynia of the upper extremities occurred in 72, toes in 6. Truncal allodynia occurred in some. Age range and age at onset of migraine were identical in allodynic and non allodynic patients. Patients with allodynia were on an average 8 years older than non-allodynic patients (p < 0.05). Correlation was found between the duration of illness as well as frequency of migraine attacks and allodynia. Frequent migraineurs exhibited prolonged allodynia. CONCLUSIONS: Clinically detectable allodynia, which occurs with high frequency in migraineurs, should become part of the clinical evaluation because of its implications in early treatment with triptans, recurrence of headache, consistency of response to triptans, and development of chronicity of migraine. PMID- 15365136 TI - Regional cerebral glucose metabolism in epilepsies with continuous spikes and waves during sleep. AB - BACKGROUND: Epileptic syndromes with continuous spikes and waves during sleep (CSWS) represent a wide spectrum of epileptic conditions associated with cognitive dysfunctions that have the EEG pattern of CSWS as a common feature. Reported are the results of voxel-based analyses of brain glucose metabolism performed in a group of 18 children with CSWS. METHODS: Voxel-based analyses of cerebral glucose metabolism were performed using statistical parametric mapping (SPM). First, each patient was compared with a control group and the influence of age, epileptic activity, and corticosteroid treatment on metabolic abnormalities was studied. Also, disease-related changes in the contribution of a brain area to the level of metabolic activity in another brain area were investigated using pathophysiologic interactions in groups of patients compared with the control group. RESULTS: Individual SPM analyses identified three metabolic patterns: association of hypermetabolic and hypometabolic areas, hypometabolic areas only, and normal pattern. Age and intensity of awake interictal spiking did not significantly differ in patients showing focal hypermetabolism compared with the other ones. Treatment with corticosteroids was associated with absence of focal hypermetabolism. In the group of patients with hypermetabolic areas, analyses of pathophysiologic interactions showed disease-related altered functional connectivity between the parietal and frontal cortices. CONCLUSIONS: Cerebral metabolic patterns are heterogeneous among patients with CSWS. This metabolic heterogeneity could be related to the use of corticosteroid treatment before PET. The parietofrontal altered connectivity observed in patients with hypermetabolism is interpreted as a phenomenon of remote inhibition of the frontal lobes induced by highly epileptogenic and hypermetabolic posterior cortex. PMID- 15365137 TI - Cortical stimulation study of the role of rhinal cortex in deja vu and reminiscence of memories. AB - OBJECTIVE: To study the role of perirhinal (PC) and entorhinal cortices (EC) in dreamy state symptoms (deja vu and reminiscence of scenes). These phenomena have been attributed to functional alteration of memory networks supported by the medial temporal lobes, principally involving the amygdala and hippocampus. The role of sub-hippocampal structures (EC and PC) in inducing these phenomena has not previously been addressed. METHODS: The authors studied the symptoms evoked by direct electrical stimulations of PC and EC in comparison with those obtained after stimulation of the amygdala and hippocampus. Stimulations were performed in a group of 24 patients with epilepsy, during stereoelectroencephalographic (SEEG) recordings in the setting of presurgical evaluation. All patients had electrodes that sampled the rhinal cortices, amygdala, and hippocampus. RESULTS: A total of 280 stimulations were analyzed. Entorhinal and perirhinal stimulations induced classic mesial temporal lobe responses (emotional, dysautonomic) but also more specific responses, particularly the deja vu phenomenon and reminiscence of scenes. Such deja vu or deja vecu type responses were produced proportionately more often by stimulation of the EC than by stimulation of the amygdala and hippocampus. In particular, deja vu was associated with stimulation of the EC and reminiscence of memories with PC stimulation. CONCLUSION: This study strongly suggests that experiential symptoms are largely dependent upon functional modification of the physiology of the rhinal cortices. PMID- 15365138 TI - Topiramate vs placebo in painful diabetic neuropathy: analgesic and metabolic effects. AB - BACKGROUND: Using identical methods, three simultaneous placebo-controlled trials of topiramate for painful diabetic neuropathy (PDN) did not reach significance. This independent yet concurrent placebo-controlled trial used different methods to assess topiramate efficacy and tolerability in PDN. METHODS: This 12-week, multicenter, randomized, double-blind trial included 323 subjects with PDN and pain visual analog (PVA) score of at least 40 on a scale from 0 (no pain) to 100 (worst possible pain). Topiramate (n = 214) or placebo (n = 109) was titrated to 400 mg daily or maximum tolerated dose. Short-acting rescue analgesics were permitted only during the first 6 weeks. RESULTS: Baseline characteristics were comparable between groups except for mean body weight (topiramate, 101.4 kg; placebo, 95.7 kg; p = 0.028). Twelve weeks of topiramate treatment reduced PVA scale score (from 68.0 to 46.2 mm) more effectively than placebo (from 69.1 to 54.0 mm; p = 0.038). Fifty percent of topiramate-treated subjects and 34% of placebo-treated subjects responded to treatment, defined as >30% reduction in PVA scale score (p = 0.004). Topiramate monotherapy also reduced worst pain intensity (p = 0.003 vs placebo) and sleep disruption (p = 0.020 vs placebo). Diarrhea, loss of appetite, and somnolence were the most commonly reported adverse events in the topiramate group. Topiramate reduced body weight (-2.6 vs +0.2 kg for placebo; p < 0.001) without disrupting glycemic control. CONCLUSIONS: Topiramate monotherapy reduced pain and body weight more effectively than placebo in patients with painful diabetic neuropathy. PMID- 15365139 TI - Sensory neuropathy in vasculitis: a clinical, pathologic, and electrophysiologic study. AB - BACKGROUND: Vasculitis is not usually considered as a cause of symmetric sensory neuropathy. OBJECTIVE AND METHODS: To present the clinical, pathologic, and electrophysiologic features of 17 (16%) cases of sensory neuropathy in vasculitis (SNV) among 106 cases with histologically proven vasculitic neuropathy that were collected over the last 30 years. RESULTS: In 41% of cases, SNV was found as systemic vasculitic neuropathy in association with primary vasculitic disease. The most common clinical presentation was symmetric polyneuropathy, seen in 53% of cases. The most common nerve conduction pattern was diffuse neuropathy pattern of axonal degeneration. Sural nerve biopsy was diagnostic in 88% of cases. In two cases, muscle biopsy was necessary for the definite diagnosis of vasculitis. Non systemic SNV is usually benign. Of 11 patients followed for longer than 2 years, none developed motor weakness due to neuropathy. CONCLUSION: Sensory neuropathy, regardless of symmetry, can be due to vasculitis. PMID- 15365140 TI - Plantar nerve AP and skin biopsy in sensory neuropathies with normal routine conduction studies. AB - OBJECTIVE: To assess the medial plantar nerve action potential (NAP) and skin biopsy in the evaluation of suspected distal sensory neuropathies (SN) with normal routine nerve conduction studies (NCS). METHODS: A total of 110 consecutive patients with suspected distal SN and normal routine NCS underwent medial plantar NAP testing and punch skin biopsy. Patients were clinically stratified as having pure small fiber sensory neuropathy (SFSN), or distal SN with large fiber involvement (SN-LFI). RESULTS: A total of 56 patients were classified as SN-LFI and 54 SFSN. The medial plantar NAP, a measure of large fiber function, was abnormal in 31.8% of patients, more frequently in SN-LFI than SFSN. Distal leg epidermal nerve fiber (ENF) density, a measure of small fibers, was reduced in 47.3% of biopsies, with isolated ENF morphologic changes in 29.1% and normal findings in 23.6%. Biopsy abnormalities were more severe and prevalent in SN-LFI than in SFSN. In patients with a normal medial plantar NAP, distal leg biopsy showed reduced ENF density in 34.7%, and isolated morphologic changes in a further 37% of cases. CONCLUSIONS: The medial plantar nerve action potential and skin biopsy are complementary in evaluation of distal SN with normal routine NCS. Small sensory nerve fibers are affected early in SN, and more severely so when large fiber involvement is apparent clinically. PMID- 15365141 TI - Homocysteine and levodopa: should Parkinson disease patients receive preventative therapy? AB - Epidemiologic evidence has linked elevation of serum homocysteine to an increased risk of coronary artery disease, stroke, and dementia. An increase in homocysteine levels in Parkinson disease (PD) recently has been discovered. Although B vitamin status and genetic factors are important modifying influences determining the degree of this elevation, the main cause appears to be therapy with L-dopa. It has been suggested that breakdown of L-dopa by catechol-O methyltransferase results in increased homocysteine formation. Therefore, there are reasons to suggest that management of PD may render patients at increased risk of stroke, heart disease, dementia, and even accelerated nigral degeneration. At present, no controlled prospective studies have evaluated this phenomenon, although they are ongoing. PMID- 15365142 TI - "Spaghetti in brain": DAVF. PMID- 15365143 TI - Two novel mutations of the glycine receptor gene in a Taiwanese hyperekplexia family. AB - The authors report a Taiwanese family with autosomal recessive hyperekplexia. Two novel mutations, W96C (from the paternal allele) and R344X (from the maternal allele), which are located in exon 4 and exon 7 of the GLRA1 gene, were identified in this family. A series of electrophysiologic investigations were conducted in one of the probands, and the results suggest that the "startle center" is located subcortically. PMID- 15365144 TI - Elevation of myoinositol is associated with disease containment in progressive multifocal leukoencephalopathy. AB - MRI, 1H-MR spectroscopy, immunologic, and virologic studies were performed in search of prognostic factors of disease evolution in patients with progressive multifocal leukoencephalopathy (PML). Acute lesions of PML survivors showed twofold higher standard score of the ratio of myoinositol (a glial marker) to creatine compared with lesions in patients whose disease progressed. Concomitantly, JC virus-specific cytotoxic T lymphocytes were only detected in the blood of PML survivors. These results suggest that inflammation limits disease progression. PMID- 15365145 TI - Salvage therapy for primary CNS lymphoma with a combination of rituximab and temozolomide. AB - The authors evaluated the efficacy of a combination of rituximab and temozolomide for recurrent or refractory primary CNS lymphoma (PCNSL). Fifteen patients with a median age of 69 years had a 53% objective response rate with acceptable toxicity. Median overall survival is 14 months and median progression free survival of responding patients is 7.7 months. This combination merits further study and provides a reasonable therapeutic alternative for older patients with progressive PCNSL. PMID- 15365146 TI - Salvage PCV chemotherapy for temozolomide-resistant oligodendrogliomas. AB - The authors investigated the results of PCV chemotherapy within a cohort of 24 patients treated within the EORTC study 26971 on temozolomide chemotherapy in recurrent oligodendroglioma. The genotype of the tumors was assessed with fluorescent in situ hybridization with locus specific probes for the region 1p36. Four of the 24 patients responded (17%). Fifty percent of patients were still free from progression at 6 months and 21% were free from progression at 12 months. Although a clear relation existed between loss of 1p and response to temozolomide chemotherapy, this relation was absent in salvage PCV chemotherapy. PMID- 15365147 TI - Thalamic deep brain stimulation activates the cerebellothalamocortical pathway. AB - To investigate the mechanism of action of deep brain stimulation (DBS), the authors studied the effects of thalamic DBS on the cerebellothalamocortical (CTC) pathway. With DBS turned off, excitability of the CTC pathway was reduced. Turning DBS on resulted in facilitation of the CTC pathway. Therefore, thalamic DBS appears to activate rather than inhibit the target area. PMID- 15365148 TI - A family with Alzheimer disease and strokes associated with A713T mutation of the APP gene. AB - Three members of an Italian family with autosomal dominant dementia and multiple strokes had the A713T mutation of the APP gene. The neuropathologic examination of the proband disclosed Alzheimer disease (AD) with severe cerebral amyloid angiopathy and multiple infarcts. This indicates that the A713T mutation of the APP gene, lying at the gamma-secretase cleavage site, can be responsible for AD with symptomatic cerebral amyloid angiopathy. PMID- 15365149 TI - APOE-dependent PET patterns of brain activation in Alzheimer disease. AB - Using H2(15)O PET, the authors imaged 13 patients with Alzheimer disease (AD) while performing a serial nonverbal recognition memory task. Patterns of brain activation differed as a function of APOE genotype: epsilon4 carriers exhibited lower activation in the left lingual gyrus and higher activation in left cuneus, precuneus, parahippocampal, and right precentral gyrus. The APOE genotype seems to play a role in cerebral physiologic activity even after onset of clinical manifestations of AD. PMID- 15365150 TI - Cerebral processing in the minimally conscious state. AB - We studied a patient in a minimally conscious state using PET and cognitive evoked potentials. Cerebral metabolism was below half of normal values. Auditory stimuli with emotional valence (infant cries and the patient's own name) induced a much more widespread activation than did meaningless noise; the activation pattern was comparable with that previously obtained in controls. Cognitive potentials showed preserved P300 responses to the patient's own name. PMID- 15365151 TI - The prevalence of pain in multiple sclerosis: a multicenter cross-sectional study. AB - In a multicenter cross-sectional study, the authors assessed pain in patients with multiple sclerosis (MS) using a symptom-oriented approach. Out of 2,077 questionnaires, we used 1,672 for data analysis. Pain and frequencies included trigeminal neuralgia 2%, Lhermitte's sign 9%, dysesthetic pain 18.1%, back pain 16.4%, and painful tonic spasms 11%. Comparison between different groups showed significant differences for age, Expanded Disability Status Scale, disease duration, and disease course, but not for sex. This study underlines the relevance of pain in the clinical history of MS. PMID- 15365152 TI - Infantile neuroaxonal dystrophy and pantothenate-kinase-associated neurodegeneration: locus heterogeneity. AB - Common clinical, radiologic, and pathologic features in infantile neuroaxonal dystrophy (INAD) and pantothenate kinase-associated neurodegeneration (PKAN) have led to the hypothesis of an allelic relationship. With the discovery of the gene defect in PKAN, this can now be tested directly. The authors excluded linkage in one consanguineous INAD family by haplotype analysis. Moreover, sequencing in seven INAD families revealed no mutations in PANK2 or in other genes of CoA biogenesis. Thus, INAD and PKAN are genetically heterogeneous disorders. PMID- 15365153 TI - "Retinal diplopia" associated with macular wrinkling. AB - Binocular diplopia is usually due to misalignment from ocular motor dysfunction. Three patients with chronic binocular vertical diplopia and small comitant hypotropic deviations due to macular displacement (heterotopia) associated with monocular retinal wrinkling are described. This maculopathy was idiopathic in two and secondary to treated retinal detachment in the third. Displacement of the macula by vitreoretinal traction is an unusual cause of binocular diplopia that requires careful ophthalmoscopy to diagnose. PMID- 15365155 TI - Long-term effect of CPAP in the treatment of nocturnal stridor in multiple system atrophy. AB - The authors found that continuous positive airway pressure (CPAP) eliminated video-polysomnographic documented stridor in 13 multiple system atrophy (MSA) subjects with vocal cord abnormalities. Long-term follow-up showed high CPAP tolerance, no recurrence of stridor, no major side effects, subjective improvement in sleep quality, and that median survival time was similar to a group of 26 MSA patients without stridor. This study shows that in MSA, CPAP is an effective noninvasive long-term therapy for nocturnal stridor. PMID- 15365154 TI - Aprataxin gene mutations in Tunisian families. AB - The authors report clinical and genetic study of 13 patients from three unrelated Tunisian families with an early onset cerebellar ataxia associated with oculomotor apraxia. Cerebellar ataxia with oculomotor apraxia 1 (AOA1) represents a clinically heterogeneous disease caused by mutations in the aprataxin gene. Two novel mutations were identified, the complete deletion of the gene, which seems to not correlate with an increased severity of the disease, and a splice mutation on the acceptor splice site of exon 7. PMID- 15365156 TI - Clinical picture of bilateral vestibular schwannomas, sudden bilateral hearing loss, and aviation. PMID- 15365157 TI - Hyperventilation and vomiting due to posterior circulation stroke. PMID- 15365158 TI - Effect of D-penicillamine on neuromuscular junction in patients with Wilson disease. PMID- 15365159 TI - Mutations in the FGF14 gene are not a major cause of spinocerebellar ataxia in Caucasians. PMID- 15365160 TI - Action observation and execution: intracranial recordings in a human subject. PMID- 15365161 TI - Acute bowler's thumb. PMID- 15365162 TI - Vitamin D intake and incidence of multiple sclerosis. PMID- 15365163 TI - Huntington's disease-like 2 can present as chorea-acanthocytosis. PMID- 15365164 TI - Ophthalmologic features of Parkinson's disease. PMID- 15365165 TI - Proneness to psychological distress is associated with risk of Alzheimer's disease. PMID- 15365166 TI - Lateralizing value of Todd's palsy in patients with epilepsy. PMID- 15365167 TI - Depression but not seizure frequency predicts quality of life in treatment resistant epilepsy. PMID- 15365168 TI - Syncope during EEG recording. PMID- 15365169 TI - Heregulin ameliorates the dystrophic phenotype in mdx mice. AB - Duchenne's muscular dystrophy (DMD) is a fatal neuromuscular disease caused by absence of dystrophin. Utrophin is a chromosome 6-encoded dystrophin-related protein (DRP), sharing functional motifs with dystrophin. Utrophin's ability to compensate for dystrophin during development and when transgenically overexpressed has provided an important impetus for identifying activators of utrophin expression. The utrophin promoter A is transcriptionally regulated in part by heregulin-mediated, extracellular signal-related kinase-dependent activation of the GABP(alpha/beta) transcription factor complex. Therefore, this pathway offers a potential mechanism to modulate utrophin expression in muscle. We tested the ability of heregulin to improve the dystrophic phenotype in the mdx mouse model of DMD. Intraperitoneal injections of a small peptide encoding the epidermal growth factor-like region of heregulin ectodomain for 3 months in vivo resulted in up-regulation of utrophin, a marked improvement in the mechanical properties of muscle as evidenced by resistance to eccentric contraction mediated damage, and a reduction of muscle pathology. The amelioration of dystrophic phenotype by heregulin-mediated utrophin up-regulation offers a pharmacological therapeutic modality and obviates many of the toxicity and delivery issues associated with viral vector-based gene therapy for DMD. PMID- 15365170 TI - Structural basis for the interaction of Escherichia coli NusA with protein N of phage lambda. AB - The C terminus of transcription factor NusA from Escherichia coli comprises two repeat units, which bind during antitermination to protein N from phage lambda. To delineate the structural basis of the NusA-lambdaN interaction, we attempted to crystallize the NusA C-terminal repeats in complex with a lambdaN peptide (residues 34-47). The two NusA domains became proteolytically separated during crystallization, and crystals contained two copies of the first repeat unit in contact with a single lambdaN fragment. The NusA modules employ identical regions to contact the peptide but approach the ligand from opposite sides. In contrast to the alpha-helical conformation of the lambdaN N terminus in complex with boxB RNA, residues 34-40 of lambdaN remain extended upon interaction with NusA. Mutational analyses indicated that only one of the observed NusA-lambdaN interaction modes is biologically significant, supporting an equimolar ratio of NusA and lambdaN in antitermination complexes. Solution studies indicated that additional interactions are fostered by the second NusA repeat unit, consistent with known compensatory mutations in NusA and lambdaN. Contrary to the RNA polymerase alpha subunit, lambdaN binding does not stimulate RNA interaction of NusA. The results demonstrate that lambdaN serves as a scaffold to closely oppose NusA and the mRNA in antitermination complexes. PMID- 15365171 TI - Morphological homoplasy, life history evolution, and historical biogeography of plethodontid salamanders inferred from complete mitochondrial genomes. AB - The evolutionary history of the largest salamander family (Plethodontidae) is characterized by extreme morphological homoplasy. Analysis of the mechanisms generating such homoplasy requires an independent molecular phylogeny. To this end, we sequenced 24 complete mitochondrial genomes (22 plethodontids and two outgroup taxa), added data for three species from GenBank, and performed partitioned and unpartitioned Bayesian, maximum likelihood, and maximum parsimony phylogenetic analyses. We explored four dataset partitioning strategies to account for evolutionary process heterogeneity among genes and codon positions, all of which yielded increased model likelihoods and decreased numbers of supported nodes in the topologies (Bayesian posterior probability >0.95) relative to the unpartitioned analysis. Our phylogenetic analyses yielded congruent trees that contrast with the traditional morphology-based taxonomy; the monophyly of three of four major groups is rejected. Reanalysis of current hypotheses in light of these evolutionary relationships suggests that (i) a larval life history stage reevolved from a direct-developing ancestor multiple times; (ii) there is no phylogenetic support for the "Out of Appalachia" hypothesis of plethodontid origins; and (iii) novel scenarios must be reconstructed for the convergent evolution of projectile tongues, reduction in toe number, and specialization for defensive tail loss. Some of these scenarios imply morphological transformation series that proceed in the opposite direction than was previously thought. In addition, they suggest surprising evolutionary lability in traits previously interpreted to be conservative. PMID- 15365172 TI - Mixing active-site components: a recipe for the unique enzymatic activity of a telomere resolvase. AB - The ResT protein, a telomere resolvase from Borrelia burgdorferi, processes replication intermediates into linear replicons with hairpin ends by using a catalytic mechanism similar to that for tyrosine recombinases and type IB topoisomerases. We have identified in ResT a hairpin binding region typically found in cut-and-paste transposases. We show that substitution of residues within this region results in a decreased ability of these mutants to catalyze telomere resolution. However, the mutants are capable of resolving heteroduplex DNA substrates designed to allow spontaneous destabilization and prehairpin formation. These findings support the existence of a hairpin binding region in ResT, the only known occurrence outside a transposase. The combination of transposase-like and tyrosine-recombinase-like domains found in ResT indicates the use of a composite active site and helps explain the unique breakage-and reunion reaction observed with this protein. Comparison of the ResT sequence with other known telomere resolvases suggests that a hairpin binding motif is a common feature in this class of enzyme; the sequence motif also appears in the RAG recombinases. Finally, our data support a mechanism of action whereby ResT induces prehairpin formation before the DNA cleavage step. PMID- 15365173 TI - AIPL1, the protein that is defective in Leber congenital amaurosis, is essential for the biosynthesis of retinal rod cGMP phosphodiesterase. AB - Aryl hydrocarbon receptor-interacting protein-like 1 (AIPL1) is a member of the FK-506-binding protein family expressed specifically in retinal photoreceptors. Mutations in AIPL1 cause Leber congenital amaurosis, a severe early-onset retinopathy that leads to visual impairment in infants. Here we show that knockdown of AIPL1 expression in mice also produces a retinopathy but over a more extended time course. Before any noticeable pathology, there was a reduction in the level of rod cGMP phosphodiesterase (PDE) proportional to the decrease in AIPL1 expression, whereas other photoreceptor proteins were unaffected. Consistent with less PDE in rods, flash responses had a delayed onset, a reduced gain, and a slower recovery of flash responses. We suggest that AIPL1 is a specialized chaperone required for rod PDE biosynthesis. Thus loss of AIPL1 would result in a condition that phenocopies retinal degenerations in the rd mouse and in a subgroup of human patients. PMID- 15365174 TI - Disruption of ceruloplasmin and hephaestin in mice causes retinal iron overload and retinal degeneration with features of age-related macular degeneration. AB - Mechanisms of brain and retinal iron homeostasis have become subjects of increased interest after the discovery of elevated iron levels in brains of patients with Alzheimer's disease and retinas of patients with age-related macular degeneration. To determine whether the ferroxidase ceruloplasmin (Cp) and its homolog hephaestin (Heph) are important for retinal iron homeostasis, we studied retinas from mice deficient in Cp and/or Heph. In normal mice, Cp and Heph localize to Muller glia and retinal pigment epithelium, a blood-brain barrier. Mice deficient in both Cp and Heph, but not each individually, had a striking, age-dependent increase in retinal pigment epithelium and retinal iron. The iron storage protein ferritin was also increased in Cp-/-Heph-/Y retinas. After retinal iron levels had increased, Cp-/-Heph-/Y mice had age-dependent retinal pigment epithelium hypertrophy, hyperplasia and death, photoreceptor degeneration, and subretinal neovascularization, providing a model of some features of the human retinal diseases aceruloplasminemia and age-related macular degeneration. This pathology indicates that Cp and Heph are critical for CNS iron homeostasis and that loss of Cp and Heph in the mouse leads to age-dependent retinal neurodegeneration, providing a model that can be used to test the therapeutic efficacy of iron chelators and antiangiogenic agents. PMID- 15365175 TI - A new model for vitamin K-dependent carboxylation: the catalytic base that deprotonates vitamin K hydroquinone is not Cys but an activated amine. AB - Vitamin K-dependent (VKD) proteins require carboxylation for diverse functions that include hemostasis, apoptosis, and Ca(2+) homeostasis, yet the mechanism of carboxylation is not well understood. Combined biochemical and chemical studies have led to a long-standing model in which a carboxylase Cys catalytic base deprotonates vitamin K hydroquinone (KH(2)), leading to KH(2) oxygenation and Glu carboxylation. We previously identified human carboxylase Cys-99 and Cys-450 as catalytic base candidates: Both were modified by N-ethylmaleimide (NEM) and Ser substituted mutants retained partial activity, suggesting that the catalytic base is activated for increased basicity. Mutants with Cys-99 or Cys-450 substituted by Ala, which cannot ionize to function as a catalytic base, were therefore analyzed. Both single and double mutants had activity, indicating that Cys-99 and Cys-450 do not deprotonate KH(2). [(14)C]NEM modification of C99A/C450A revealed one additional reactive group; however, Ser-substituted mutants of each of the eight remaining Cys retained substantial activity. To unequivocally test, then, whether any Cys or Cys combination acts as the catalytic base, a mutant with all 10 Cys substituted by Ala was generated. This mutant showed 7% wild-type activity that depended on factor IX coexpression, indicating a VKD protein effect on carboxylase maturation. NEM and diethyl pyrocarbonate inhibition suggested that the catalytic base is an activated His. These results change the paradigm for VKD protein carboxylation. The identity of the catalytic base is critical to understanding carboxylase mechanism and this work will therefore impact both reinterpretation of previous studies and future ones that define how this important enzyme functions. PMID- 15365176 TI - A nontransgenic mouse model shows inducible amyloid-beta (Abeta) peptide deposition and elucidates the role of apolipoprotein E in the amyloid cascade. AB - The amyloid-beta (Abeta) peptide, a major pathological hallmark of Alzheimer's disease (AD), undergoes a cascade of interactions resulting in the formation of soluble aggregates and their conversion in the brain to insoluble deposits and mature senile plaques. Furthermore, the apoE4 isoform of apolipoprotein E (apoE), which is the major genetic risk factor of AD, is associated with increased Abeta deposition. It is not known how the different Abeta aggregates in the amyloid cascade are formed, contribute to the pathogenesis of AD, or are affected by apoE4. To investigate the initial aggregation stages underlying the amyloid cascade in vivo and how apoE affects them, we examined the effects of prolonged inhibition and subsequent reactivation of the Abeta-degrading protease neprilysin on deposition, disaggregation, and fibrillization of Abeta in apoE-transgenic and control mice. In control mice, intracerebroventricular infusion of thiorphan, which inhibits neprilysin, induced Abeta42 and Abeta40 deposition and fibrillization. On termination of thiorphan treatment, the number of Abeta deposits decreased, whereas the fibrillar Abeta deposits were unaffected. Similar treatments in apoE-deficient mice and mice transgenic for human apoE4 or apoE3 revealed that apoE4 enhances specifically the nucleation and aggregation of immunopositive Abeta deposits and that reversible disaggregation of these deposits and their irreversible conversion to fibrillar deposits are stimulated similarly by the different apoE isoforms. Deposition of Abeta and its enhancement by apoE4 were accompanied by increased astrogliosis both far from and near the Abeta deposits, suggesting that astrogliosis might be triggered by both insoluble and soluble Abeta aggregates. PMID- 15365177 TI - Dermatitis due to epiregulin deficiency and a critical role of epiregulin in immune-related responses of keratinocyte and macrophage. AB - Epidermal growth factor (EGF) family members, including epiregulin (EP), play a fundamental role in epithelial tissues; however, their roles in immune responses and the physiological role of EP remain to be elucidated. The skin has a versatile system of immune surveillance. Biologically active IL-1alpha is released to extracellular space upon damage from keratinocytes and is a major player in skin inflammation. Here, we show that EP is expressed not only in keratinocytes but also in tissue-resident macrophages, and that EP-deficient (EP( /-)) mice develop chronic dermatitis. Wound healing in the skin in EP(-/-) mice was not impaired in vivo, nor was the growth rate of keratinocytes from EP(-/-) mice different from that of WT mice in vitro. Of interest is that in WT keratinocytes, both IL-1alpha and the secreted form of EP induced down-regulation of IL-18 mRNA expression, which overexpression in the epidermis was reported to induce skin inflammation in mice, whereas the down-regulation of IL-18 induced by IL-1alpha was impaired in EP(-/-) keratinocytes. Although bone marrow transfer experiments indicated that EP deficiency in non-bone-marrow-derived cells is essential for the development of dermatitis, production of proinflammatory cytokines by EP(-/-) macrophages in response to Toll-like receptor agonists was much lower, compared with WT macrophages, whose dysfunction in EP(-/-) macrophages was not compensated by the addition of the secreted form of EP. These findings, taken together, suggested that EP plays a critical role in immune/inflammatory-related responses of keratinocytes and macrophages at the barrier from the outside milieu and that the secreted and membrane-bound forms of EP have distinct functions. PMID- 15365178 TI - Leber congenital amaurosis linked to AIPL1: a mouse model reveals destabilization of cGMP phosphodiesterase. AB - Leber congenital amaurosis (LCA4) has been linked to mutations in the photoreceptor-specific gene Aryl hydrocarbon interacting protein like 1 (Aipl1). To investigate the essential role of AIPL1 in retina, we generated a mouse model of LCA by inactivating the Aipl1 gene. In Aipl1(-/-) retinas, the outer nuclear layer develops normally, but rods and cones then quickly degenerate. Aipl1(-/-) mice have highly disorganized, short, fragmented photoreceptor outer segments and lack both rod and cone electroretinogram responses. Recent biochemical evidence indicates that AIPL1 can enhance protein farnesylation. Our study reveals that rod cGMP phosphodiesterase, a farnesylated protein, is absent and cGMP levels are elevated in AIPL1(-/-) retinas before the onset of degeneration. Our findings demonstrate that AIPL1 enhances the stability of phosphodiesterase and is essential for photoreceptor viability. PMID- 15365179 TI - Par-1 promotes a hepatic mode of apical protein trafficking in MDCK cells. AB - Simple (i.e., nonstratified) epithelial cells use two different routes to target their newly synthesized luminal plasma membrane proteins to the cell surface: a direct route from the Golgi complex, as in the kidney-derived MDCK cell line, or an indirect route that involves a intermediate stop at the ab-luminal (basolateral) membrane, as in hepatocytes. The mechanisms or proteins responsible for these different protein targeting strategies are not known. Here, we show that increased expression of EMK1, a mammalian ortholog of Caenorhabditis elegans Par-1, in MDCK cells promotes a switch from a direct to a transcytotic mode of apical protein delivery and other trafficking changes typical of hepatocytes. These results, together with our recent demonstration that PAR-1 promotes morphological features of hepatocytes in MDCK cells, indicate that Par-1 modulates the developmental decision to build a columnar versus a hepatic epithelial cell. To our knowledge, Par-1 is the first gene assigned to this task in epithelial morphogenesis. PMID- 15365180 TI - Targeted knockout in Physcomitrella reveals direct actions of phytochrome in the cytoplasm. AB - The plant photoreceptor phytochrome plays an important role in the nucleus as a regulator of transcription. Numerous studies imply, however, that phytochromes in both higher and lower plants mediate physiological reactions within the cytoplasm. In particular, the tip cells of moss protonemal filaments use phytochrome to sense light direction, requiring a signaling system that transmits the directional information directly to the microfilaments that direct tip growth. In this work we describe four canonical phytochrome genes in the model moss species Physcomitrella patens, each of which was successfully targeted via homologous recombination and the distinct physiological functions of each gene product thereby identified. One homolog in particular mediates positive phototropism, polarotropism, and chloroplast movement in polarized light. This photoreceptor thus interacts with a cytoplasmic signal/response system. This is our first step in elucidating the cytoplasmic signaling function of phytochrome at the molecular level. PMID- 15365181 TI - Selective transport systems mediate sequestration of plant glucosides in leaf beetles: a molecular basis for adaptation and evolution. AB - Chrysomeline larvae respond to disturbance and attack by everting dorsal glandular reservoirs, which release defensive secretions. The ancestral defense is based on the de novo synthesis of monoterpene iridoids. The catabolization of the host-plant O-glucoside salicin into salicylaldehyde is a character state that evolved later in two distinct lineages, which specialized on Salicaceae. By using two species producing monoterpenes (Hydrothassa marginella and Phratora laticollis) and two sequestering species (Chrysomela populi and Phratora vitellinae), we studied the molecular basis of sequestration by feeding the larvae structurally different thioglucosides resembling natural O-glucosides. Their accumulation in the defensive systems demonstrated that the larvae possess transport systems, which are evolutionarily adapted to the glycosides of their host plants. Minor structural modifications in the aglycon result in drastically reduced transport rates of the test compounds. Moreover, the ancestral iridoid producing leaf beetles already possess a fully functional import system for an early precursor of the iridoid defenses. Our data confirm an evolutionary scenario in which, after a host-plant change, the transport system of the leaf beetles may play a pivotal role in the adaptation on new hosts by selecting plant derived glucosides that can be channeled to the defensive system. PMID- 15365182 TI - Iron-oxo clusters biomineralizing on protein surfaces: structural analysis of Halobacterium salinarum DpsA in its low- and high-iron states. AB - The crystal structure of the Dps-like (Dps, DNA-protecting protein during starvation) ferritin protein DpsA from the halophile Halobacterium salinarum was determined with low endogenous iron content at 1.6-A resolution. The mechanism of iron uptake and storage was analyzed in this noncanonical ferritin by three high resolution structures at successively increasing iron contents. In the high-iron state of the DpsA protein, up to 110 iron atoms were localized in the dodecameric protein complex. For ultimate iron storage, the archaeal ferritin shell comprises iron-binding sites for iron translocation, oxidation, and nucleation. Initial iron-protein interactions occur through acidic residues exposed along the outer surface in proximity to the iron entry pore. This narrow pore permits translocation of ions toward the ferroxidase centers via two discrete steps. Iron oxidation proceeds by transient formation of tri-iron ferroxidase centers. Iron storage by biomineralization inside the ferritin shell occurs at two iron nucleation centers. Here, a single iron atom provides a structural seed for iron oxide cluster formation. The clusters with up to five iron atoms adopt a geometry that is different from natural biominerals like magnetite but resembles iron clusters so far known only from bioinorganic model compounds. PMID- 15365183 TI - Electrical detection of single viruses. AB - We report direct, real-time electrical detection of single virus particles with high selectivity by using nanowire field effect transistors. Measurements made with nanowire arrays modified with antibodies for influenza A showed discrete conductance changes characteristic of binding and unbinding in the presence of influenza A but not paramyxovirus or adenovirus. Simultaneous electrical and optical measurements using fluorescently labeled influenza A were used to demonstrate conclusively that the conductance changes correspond to binding/unbinding of single viruses at the surface of nanowire devices. pH dependent studies further show that the detection mechanism is caused by a field effect, and that the nanowire devices can be used to determine rapidly isoelectric points and variations in receptor-virus binding kinetics for different conditions. Lastly, studies of nanowire devices modified with antibodies specific for either influenza or adenovirus show that multiple viruses can be selectively detected in parallel. The possibility of large-scale integration of these nanowire devices suggests potential for simultaneous detection of a large number of distinct viral threats at the single virus level. PMID- 15365184 TI - Listeria-based cancer vaccines that segregate immunogenicity from toxicity. AB - The facultative intracellular bacterium Listeria monocytogenes is being developed as a cancer vaccine platform because of its ability to induce potent innate and adaptive immunity. For successful clinical application, it is essential to develop a Listeria platform strain that is safe yet retains the potency of vaccines based on wild-type bacteria. Here, we report the development of a recombinant live-attenuated vaccine platform strain that retains the potency of the fully virulent pathogen, combined with a >1,000-fold reduction in toxicity, as compared with wild-type Listeria. By selectively deleting two virulence factors, ActA (DeltaactA) and Internalin B (DeltainlB), the immunopotency of Listeria was maintained and its toxicity was diminished in vivo, largely by blocking the direct internalin B-mediated infection of nonphagocytic cells, such as hepatocytes, and the indirect ActA-mediated infection by cell-to-cell spread from adjacent phagocytic cells. In contrast, infection of phagocytic cells was not affected, leaving intact the ability of Listeria to stimulate innate immunity and to induce antigenspecific cellular responses. Listeria DeltaactA/DeltainlB based vaccines were rapidly cleared from mice after immunization and induced potent and durable effector and memory T-cell responses with no measurable liver toxicity. Therapeutic vaccination of BALB/c mice bearing murine CT26 colon tumor lung metastases or palpable s.c. tumors (>100 mm(3)) with recombinant Listeria DeltaactA/DeltainlB expressing an endogenous tumor antigen resulted in breaking of self-tolerance and long-term survival. We propose that recombinant Listeria DeltaactA/DeltainlB expressing human tumor-associated antigens represents an attractive therapeutic strategy for further development and testing in human clinical trials. PMID- 15365185 TI - Folate biofortification in tomatoes by engineering the pteridine branch of folate synthesis. AB - Plants are the main source of folate in human diets, but many fruits, tubers, and seeds are poor in this vitamin, and folate deficiency is a worldwide problem. Plants synthesize folate from pteridine, p-aminobenzoate (PABA), and glutamate moieties. Pteridine synthesis capacity is known to drop in ripening tomato fruit; therefore, we countered this decline by fruit-specific overexpression of GTP cyclohydrolase I, the first enzyme of pteridine synthesis. We used a synthetic gene based on mammalian GTP cyclohydrolase I, because this enzyme is predicted to escape feedback control in planta. This engineering maneuver raised fruit pteridine content by 3- to 140-fold and fruit folate content by an average of 2 fold among 12 independent transformants, relative to vector-alone controls. Most of the folate increase was contributed by 5-methyltetrahydrofolate polyglutamates and 5,10-methenyltetrahydrofolate polyglutamates, which were also major forms of folate in control fruit. The accumulated pteridines included neopterin, monapterin, and hydroxymethylpterin; their reduced forms, which are folate biosynthesis intermediates; and pteridine glycosides not previously found in plants. Engineered fruit with intermediate levels of pteridine overproduction attained the highest folate levels. PABA pools were severely depleted in engineered fruit that were high in folate, and supplying such fruit with PABA by means of the fruit stalk increased their folate content by up to 10-fold. These results demonstrate that engineering a moderate increase in pteridine production can significantly enhance the folate content in food plants and that boosting the PABA supply can produce further gains. PMID- 15365186 TI - In situ analysis of repair processes for oxidative DNA damage in mammalian cells. AB - Oxidative DNA damage causes blocks and errors in transcription and replication, leading to cell death and genomic instability. Although repair mechanisms of the damage have been extensively analyzed in vitro, the actual in vivo repair processes remain largely unknown. Here, by irradiation with an UVA laser through a microscope lens, we have conditionally produced single-strand breaks and oxidative base damage at restricted nuclear regions of mammalian cells. We showed, in real time after irradiation by using antibodies and GFP-tagged proteins, rapid and ordered DNA repair processes of oxidative DNA damage in human cells. Furthermore, we characterized repair pathways by using repair-defective mammalian cells and found that DNA polymerase beta accumulated at single-strand breaks and oxidative base damage by means of its 31- and 8-kDa domains, respectively, and that XRCC1 is essential for both polymerase beta-dependent and proliferating cell nuclear antigen-dependent repair pathways of single-strand breaks. Thus, the repair of oxidative DNA damage is based on temporal and functional interactions among various proteins operating at the site of DNA damage in living cells. PMID- 15365187 TI - De novo synthesis and development of an RNA enzyme. AB - Arbitrary manipulation of molecular recognition at the atomic level has many applications. However, systematic design and de novo synthesis of an artificial enzyme based on such manipulation has been a long-standing challenge in the field of chemistry and biotechnology. In this report, we developed an artificial RNA ligase by implementing a synthetic strategy that fuses a series of 3D molecular modelings based on naturally occurring RNA-RNA recognition motifs with a small scale combinatorial synthesis of a modular catalytic unit. The resulting ligase produces a 3'-5' linkage in a template-directed manner for any combinations of two nucleotides at the reaction site. The reaction rate is 10(6)-fold over that of the uncatalyzed reaction with a yield higher than those of previously reported ligase ribozymes. The strategy may be applicable to the synthesis and development of a variety of nonnatural functional RNAs with defined 3D structures. PMID- 15365188 TI - Induction of WT1 (Wilms' tumor gene)-specific cytotoxic T lymphocytes by WT1 peptide vaccine and the resultant cancer regression. AB - The Wilms' tumor gene WT1 is overexpressed in leukemias and various types of solid tumors, and the WT1 protein was demonstrated to be an attractive target antigen for immunotherapy against these malignancies. Here, we report the outcome of a phase I clinical study of WT1 peptide-based immunotherapy for patients with breast or lung cancer, myelodysplastic syndrome, or acute myeloid leukemia. Patients were intradermally injected with an HLA-A*2402-restricted, natural, or modified 9-mer WT1 peptide emulsified with Montanide ISA51 adjuvant at 0.3, 1.0, or 3.0 mg per body at 2-week intervals, with toxicity and clinical and immunological responses as the principal endpoints. Twenty-six patients received one or more WT1 vaccinations, and 18 of the 26 patients completed WT1 vaccination protocol with three or more injections of WT1 peptides. Toxicity consisted only of local erythema at the WT1 vaccine injection sites in patients with breast or lung cancer or acute myeloid leukemia with adequate normal hematopoiesis, whereas severe leukocytopenia occurred in patients with myelodysplastic syndrome with abnormal hematopoiesis derived from WT1-expressing, transformed hematopoietic stem cells. Twelve of the 20 patients for whom the efficacy of WT1 vaccination could be assessed showed clinical responses such as reduction in leukemic blast cells or tumor sizes and/or tumor markers. A clear correlation was observed between an increase in the frequencies of WT1-specific cytotoxic T lymphocytes after WT1 vaccination and clinical responses. It was therefore demonstrated that WT1 vaccination could induce WT1-specific cytotoxic T lymphocytes and result in cancer regression without damage to normal tissues. PMID- 15365189 TI - Imidazoleacetic acid-ribotide: an endogenous ligand that stimulates imidazol(in)e receptors. AB - We identified the previously unknown structures of ribosylated imidazoleacetic acids in rat, bovine, and human tissues to be imidazole-4-acetic acid-ribotide (IAA-RP) and its metabolite, imidazole-4-acetic acid-riboside. We also found that IAA-RP has physicochemical properties similar to those of an unidentified substance(s) extracted from mammalian tissues that interacts with imidazol(in)e receptors (I-Rs). ["Imidazoline," by consensus (International Union of Pharmacology), includes imidazole, imidazoline, and related compounds. We demonstrate that the imidazole IAA-RP acts at I-Rs, and because few (if any) imidazolines exist in vivo, we have adopted the term "imidazol(in)e-Rs."] The latter regulate multiple functions in the CNS and periphery. We now show that IAA RP (i) is present in brain and tissue extracts that exhibit I-R activity; (ii) is present in neurons of brainstem areas, including the rostroventrolateral medulla, a region where drugs active at I-Rs are known to modulate blood pressure; (iii) is present within synaptosome-enriched fractions of brain where its release is Ca(2+)-dependent, consistent with transmitter function; (iv) produces I-R-linked effects in vitro (e.g., arachidonic acid and insulin release) that are blocked by relevant antagonists; and (v) produces hypertension when microinjected into the rostroventrolateral medulla. Our data also suggest that IAA-RP may interact with a novel imidazol(in)e-like receptor at this site. We propose that IAA-RP is a neuroregulator acting via I-Rs. PMID- 15365190 TI - Celebrating a decade of neuroscience databases: looking to the future of high throughput data analysis, data integration, and discovery neuroscience. PMID- 15365191 TI - A gateway to the future of neuroinformatics. PMID- 15365192 TI - Scaling self-organizing maps to model large cortical networks. AB - Self-organizing computational models with specific intracortical connections can explain many functional features of visual cortex, such as topographic orientation and ocular dominance maps. However, due to their computational requirements, it is difficult to use such detailed models to study large-scale phenomenal like object segmentation and binding, object recognition, tilt illusions, optic flow, and fovea-periphery differences. This article introduces two techniques that make large simulations practical. First, we show how parameter scaling equations can be derived for laterally connected self organizing models. These equations result in quantitatively equivalent maps over a wide range of simulation sizes, making it possible to debug small simulations and then scale them up only when needed. Parameter scaling also allows detailed comparison of biological maps and parameters between individuals and species with different brain region sizes. Second, we use parameter scaling to implement a new growing map method called GLISSOM, which dramatically reduces the memory and computational requirements of large self-organizing networks. With GLISSOM, it should be possible to simulate all of human V1 at the single-column level using current desktop workstations. We are using these techniques to develop a new simulator Topographica, which will help make it practical to perform detailed studies of large-scale phenomena in topographic maps. PMID- 15365193 TI - Classification of HIV-1-mediated neuronal dendritic and synaptic damage using multiple criteria linear programming. AB - The ability to identify neuronal damage in the dendritic arbor during HIV-1 associated dementia (HAD) is crucial for designing specific therapies for the treatment of HAD. To study this process, we utilized a computer-based image analysis method to quantitatively assess HIV-1 viral protein gp120 and glutamate mediated individual neuronal damage in cultured cortical neurons. Changes in the number of neurites, arbors, branch nodes, cell body area, and average arbor lengths were determined and a database was formed (http://dm.ist.unomaha. edu/database.htm). We further proposed a two-class model of multiple criteria linear programming (MCLP) to classify such HIV-1-mediated neuronal dendritic and synaptic damages. Given certain classes, including treatments with brain-derived neurotrophic factor (BDNF), glutamate, gp120 or non-treatment controls from our in vitro experimental systems, we used the two-class MCLP model to determine the data patterns between classes in order to gain insight about neuronal dendritic damages. This knowledge can be applied in principle to the design and study of specific therapies for the prevention or reversal of neuronal damage associated with HAD. Finally, the MCLP method was compared with a well-known artificial neural network algorithm to test for the relative potential of different data mining applications in HAD research. PMID- 15365194 TI - Semi-automated population of an online database of neuronal models (ModelDB) with citation information, using PubMed for validation. AB - Citations play an important role in medical and scientific databases by indicating the authoritative source of the data. Manual citation entry is tedious and prone to errors. We describe a method and make available computer scripts which automate the process of citation entry. We use an open citation project PERL module (PARSER) for parsing citation data that is then used to retrieve PubMed records to supply the (validated) reference. Our PERL scripts are available via a link in the web references section of this article. PMID- 15365195 TI - Functional holography of recorded neuronal networks activity. AB - We present a new approach for analyzing multi-channel recordings, such as ECoG (electrocorticograph) recordings of cortical brain activity and of individual neuron dynamics, in cultured networks. The latter are used here to illustrate the method and its ability to discover hidden functional connectivity motifs in the recorded activity. The cultured networks are formed from dissociated mixtures of cortical neurons and glia-cells that are homogeneously spread over multi electrode array for recording of the neuronal activity. Rich, spontaneous dynamical behavior is detected, marked by the formation of temporal sequences of synchronized bursting events (SBEs), partitioned into statistically distinguishable subgroups, each with its own characteristic spatio-temporal pattern of activity.In analogy with coherence connectivity networks for multi location cortical recordings, we evaluated the inter-neuron correlation-matrix for each subgroup. Ordinarily such matrices are mapped onto a connectivity network between neuron positions in real space. In our functional holography, the correlations are normalized by the correlation distances Euclidian distances between the matrix columns. Then, we project the N-dimensional (for N channels) space spanned by the matrix of the normalized correlations, or correlation affinities, onto a corresponding 3D manifold (3D Cartesian space constructed by the three leading principal vectors of the principal component algorithm). The neurons are located by their principal eigenvalues and linked by their original (not normalized) correlations. By looking at these holograms, hidden causal motifs are revealed: each SBEs subgroup generates its characteristic connectivity diagram (network) in the 3D manifold, where the neuron locations and their links form simple structures. Moreover, the computed temporal ordering of neuron activity, when projected onto the connectivity diagrams, also exhibits simple patterns of causal propagation. We show that the method can expose functional connectivity motifs like the co-existence of subneuronal functional networks in the space of affinities. The method can be directly utilized to construct similar causal holograms for recorded brain activity. We expect that by doing so, hidden functional connectivity motifs with relevance to the understanding of brain activity might be discovered. PMID- 15365196 TI - Clustered organization of cortical connectivity. AB - Long-range corticocortical connectivity in mammalian brains possesses an intricate, nonrandom organization. Specifically, projections are arranged in "small-world" networks, forming clusters of cortical areas, which are closely linked among each other, but less frequently with areas in other clusters. In order to delineate the structure of cortical clusters and identify their members, we developed a computational approach based on evolutionary optimization. In different compilations of connectivity data for the cat and macaque monkey brain, the algorithm identified a small number of clusters that broadly agreed with functional cortical subdivisions. We propose a simple spatial growth model for evolving clustered connectivity, and discuss structural and functional implications of the clustered, small-world organization of cortical networks. PMID- 15365197 TI - Learning to predict through adaptation. AB - Common themes underlying three recent studies of mine on disparate topics are reviewed: the lamination of sensory cortex; the differentiation into subfields of the mammalian hippocampus; and the neuronal dynamics that might underlie the faculty for language in the human frontal lobes. These studies all discuss the evolution of cortical networks in terms of their computations, quantified by simulating simplified formal models. They all dwell on the interrelationship between qualitative and quantitative change. Finally, they all include, as a necessary ingredient of the relevant computational mechanism, a simple feature of pyramidal cell biophysics: firing rate adaptation. PMID- 15365198 TI - Role of pancreatic stents in prevention of post-ERCP pancreatitis. PMID- 15365199 TI - Vascular complications of pancreatitis. PMID- 15365200 TI - Pancreatic pseudocysts following acute pancreatitis: risk factors influencing therapeutic outcomes. AB - CONTEXT: The natural history of pancreatic pseudocysts has become well known in recent years, but the choice of a proper treatment still remains controversial. OBJECTIVE: This study aims at establishing whether predictive factors influencing therapeutic outcomes exist. SETTING: Patients with pancreatic pseudocysts following an episode of acute pancreatitis treated from January 1980 to December 2001 at the Department of General Surgery and Organ Transplantation of the University of Parma, Italy. PATIENTS: Seventy-four patients were studied: 12 had a spontaneous resolution, 37 patients were treated surgically, 15 were treated endoscopically and in 10, percutaneous drainage was used. MAIN OUTCOME MEASURES: Epidemiological, clinical and pathological characteristics of patients with pancreatic pseudocysts were related to morbidity, recurrence rates and hospital stay. RESULTS: At univariate logistic regression, our data reveal a significant increase in morbidity related to age (P=0.013), etiology (alcoholic vs. biliary, P=0.024), Ranson score of previous pancreatitis (P=0.006), nutritional assessment (P=0.001), residual necrosis (P<0.001) and modality of treatment (P=0.009), whereas none of these parameters has been shown to be significantly correlated to recurrence. At multivariate logistic regression, only residual necrosis was significantly related to morbidity. CONCLUSIONS: Some factors, such as epidemiological (age, etiology), clinical (severity of previous pancreatitis, malnourishment), pathological (residual necrosis), and therapeutical factors (emergency/urgency treatment) are predictive of worse outcomes for invasive treatment of pseudocysts. In particular residual necrosis appeared to be the most important factor influencing invasive treatment outcomes, confirming that this pathological aspect deserves particular attention from surgeons. No risk factors predicting pancreatic pseudocyst recurrence emerged. PMID- 15365201 TI - Solid pseudopapillary tumor as a possible cause of acute pancreatitis. AB - CONTEXT: Acute pancreatitis is not commonly seen in the first presentation of pancreatic neoplasms. Solid pseudopapillary tumor as a cause of acute pancreatitis has not yet been reported. This is the first report of acute pancreatitis resulting from solid pseudopapillary tumor. CASE REPORT: We report the case of a 21-year-old female who presented with a sudden onset of severe abdominal pain associated with elevated serum pancreatic enzyme concentration. The initial diagnosis was acute pancreatitis. However, subsequent ultrasonography and computed tomography showed an abdominal mass in the tail of the pancreas, retroperitoneal fluid and left pleural effusion. There was scarce pain relief even with large doses of analgesics. A distal pancreatectomy was then performed and a final diagnosis of solid pseudopapillary tumor was made histologically. The surrounding pancreatic tissue was characterized as hemorrhagic edematous pancreatitis. CONCLUSIONS: Solid pseudopapillary tumor is generally known as a slow-growing pancreatic neoplasm with few, if any, symptoms. However, solid pseudopapillary tumors should be kept in mind as a possible cause of acute pancreatitis, especially in cases of non-alcoholic young women having an acute pancreatitis attack. PMID- 15365202 TI - Estrogen-induced severe acute pancreatitis in a male. AB - CONTEXT: Acute pancreatitis is related to drugs in 1.4-2% of cases. Estrogens are an uncommon but well-known risk factor of pancreatitis in women and men with pre existing hyperlipidemia. CASE REPORT: We report the case of a 37-year-old man with covert hypertriglyceridemia who developed a severe life-threatening pancreatitis strongly associated with estrogen therapy preparatory to sex change surgery, characterized by a massive triglyceride level, pancreatic insufficiency and multiple organ failure at the time of the diagnosis. Other causes of the disease were ruled out. CONCLUSIONS: To our knowledge, this is the first description of severe necrotizing estrogen-induced pancreatitis in a male. Baseline abnormal triglyceride levels should be checked by physicians before starting estrogen therapy in women and men. PMID- 15365203 TI - The scrotum in pancreatitis: a case report and literature review. AB - CONTEXT: An acute inguinoscrotal swelling appearing during the course of acute pancreatitis is rare. There are only three case reports of this in the English literature. CASE REPORT: We report a case of right inguinoscrotal swelling appearing during an attack of acute pancreatitis. CONCLUSIONS: A correct diagnosis and appropriate management will prevent unnecessary surgical intervention. PMID- 15365204 TI - Inflammatory myofibroblastic tumor presenting as a pancreatic mass: a case report and review of the literature. AB - CONTEXT: Inflammatory myofibroblastic tumor is a distinctive lesion of unknown etiology. It has generally been considered a rare benign pseudosarcomatous lesion of admixed inflammatory infiltrates with myofibroblastic spindle cells. Although original case descriptions focused on the pulmonary system, it is now recognized that virtually any anatomic location can be involved. However, an inflammatory myofibroblastic tumor located in the pancreas is rare. CASE REPORT: We report a case of an asymptomatic 70-year-old Caucasian man with a 3.8 cm inflammatory myofibroblastic tumor located in the tail of the pancreas which was discovered incidentally on a computed tomography scan of the abdomen. Endoscopic ultrasonography with fine needle aspiration was negative for malignancy. However, because of worrisome radiographic features, a distal pancreatectomy with splenectomy was performed. The pathology revealed an inflammatory myofibroblastic tumor with focal extension into the peripancreatic soft tissues, but with negative surgical margins. The patient has been followed for 10 months without evidence of recurrence. CONCLUSIONS: To date, there have been only 25 cases of inflammatory myofibroblastic tumor located in the pancreas reported in the English language scientific literature. Even with multimodal pre-surgical investigation, it can be extremely difficult to differentiate inflammatory myofibroblastic tumor from pancreatic malignancies. Most cases require surgical exploration and complete resection to obtain an accurate diagnosis. A review of published case reports is also presented. PMID- 15365205 TI - A case report of the unusual presence of hydatid disease in the pancreas and breast. AB - CONTEXT: Hydatid cyst disease can occur in all viscera and soft tissues although the liver and the lungs are the most commonly involved organs. Pancreatic and breast involvement with the disease are extremely rare and the presence of this disease in these organs without any other organ involvement has not been previously reported. CASE REPORT: We present herein a patient with isolated pancreatic cyst hydatid treated successfully with complete cyst excision. However, 18 months after the pancreatic surgery, hydatid disease was found in the breast and again the cyst was completely removed. No other organ involvement has been detected during a 2-year follow-up. CONCLUSIONS: Hydatid disease should be considered in the differential diagnosis of all cystic masses in all anatomic locations, especially in regions where the disease is endemic. The ideal treatment is the complete excision of the cyst without any spillage. PMID- 15365206 TI - Hemosuccus pancreaticus: an uncommon cause of gastrointestinal hemorrhage. A case report. AB - CONTEXT: Pseudoaneurysms of peripancreatic arteries usually arise as a complication of acute and chronic pancreatitis. These pseudoaneurysms may either bleed intra abdominally following rupture or may erode into the adjacent hollow viscera and manifest as gastrointestinal bleeding. Pseudoaneurysms rarely communicate with the pancreatic duct and bleeding occurs from the ampulla of Vater in the form of hemosuccus pancreaticus. CASE REPORT: We report a patient with chronic pancreatitis who presented with hemosuccus pancreaticus. Contrast enhanced computerized tomography and angiography revealed bleeding from a left gastric artery pseudoaneurysm. Despite multiple attempts, embolization was unsuccessful due to arterial spasm. The patient was successfully managed at emergency surgery by excision of the pseudoaneurysm sac and ligation of the left gastric artery. CONCLUSION: Bleeding from visceral artery pseudoaneurysms rarely manifests itself as hemosuccus pancreaticus. Most cases can be managed by angio embolization. However, in patients with recurrent bleeding or failed embolization, emergency surgery is required. PMID- 15365207 TI - Pancreaticoduodenectomy for pancreatic metastases from breast carcinoma. AB - CONTEXT: Unlike primary pancreatic carcinoma, metastases to the pancreas are rare events and their resection may be performed as a palliative treatment. The aim of this study was to review our experience with the operative management of pancreatic metastases from breast carcinoma. CASE REPORT: Three patients underwent pancreaticoduodenectomy for pancreatic metastases from breast carcinoma between 1990 and 2002. The clinical features of and the results after resection were examined. The presenting clinical symptoms were jaundice, pain and weight loss. In all cases, the pancreatic metastases were solitary lesions located in the head of the pancreas and the intervals between the primary breast carcinoma and the pancreatic metastases were 3, 5 and 7 years. The primary tumor was lobular carcinoma. Metastases were initially misdiagnosed as primary cancers. All patients underwent pancreaticoduodenectomy. One patient died from disseminated metastatic disease 26 months after surgery. The other two patients are alive 37 and 21 months after pancreaticoduodenectomy; the second one has had recurrence of the disease. CONCLUSION: In rare cases, the pancreas may be the presenting site for breast metastases, typically years after treating the primary tumor. Pancreaticoduodenectomy can be performed in the absence of extreme metastatic disease; it can provide worthwhile palliation and improve survival. PMID- 15365208 TI - Coxsackievirus infection associated with acute pancreatitis. AB - CONTEXT: A variety of infectious agents have been reported as rare causes of acute pancreatitis. CASE REPORT: We briefly describe a 36-year-old man who presented with acute pancreatitis and a maculopapular rash. The marked elevation in antibody titer against coxsackievirus B, as well as the skin biopsy, was compatible with acute coxsackievirus B viral infection. CONCLUSION: This case highlights the fact that an appropriate investigation for viral infections should be performed in patients having acute pancreatitis and no classical risk factors. PMID- 15365209 TI - Olanzapine-induced pancreatitis: a case report. AB - CONTEXT: The antipsychotic agent clozapine has been linked to several cases of pancreatitis. The newer, but related, olanzapine was believed to have fewer side effects. CASE REPORT: A 42-year-old man in good physical condition gradually developed hypertriglyceridemia, hypercholesterolemia, elevated alanine aminotransferase, diabetes and, ultimately, acute pancreatitis after 19 months of olanzapine monotherapy. Due to multiorgan failure, he was in the intensive care unit and surgical ward for 29 days. He made a full recovery. The olanzapine was discontinued. Glucose, triglyceride and cholesterol levels normalized as did pancreas and liver function. CONCLUSIONS: We report olanzapine as the probable cause of acute pancreatitis in a patient without any known predisposing factors. Olanzapine-treated patients should be monitored with glucose, lipid, pancreatic function and liver function tests, and the olanzapine should be discontinued if the results of these tests worsen. PMID- 15365210 TI - Lymphoepithelial cyst: a rare cystic tumor of the pancreas which mimics carcinoma. PMID- 15365211 TI - Hepatic and splenic arteries' pseudoaneurysms causing hemosuccus pancreaticus. PMID- 15365212 TI - Pancreatic cancer: the effect of specialization and new medical treatment. PMID- 15365213 TI - Acetate-free hemodialysis: a feasibility study on a technical alternative to bicarbonate dialysis. AB - This study aimed at evaluating the feasibility of an acetate-free hemodialysis (AFHD) technique, comparing it with acetate-free biofiltration (AFB) and bicarbonate dialysis (BD). The assessment of the parameters concerned: electrolyte kinetics (Na+, K+), acid-base balance (HCO3-, pH), dialysis efficiency (Kt/V), serum beta2-microglobulin reduction ratio, nutritional status (normalized protein catabolic rate, serum albumin and total proteins, body mass index), hemopoietic status (hemoglobin, hematocrit), and some clinical parameters (systolic and diastolic blood pressures, heart rate, percent blood volume reduction measured by Hemoscan). Nine patients participated in this study which was conducted using a Latin square randomized experimental design. The results of the last week of each month of the study (1 month for each technique) were analyzed by means of Anova for repeated measures. The different treatments were comparable with regard to the main dialysis parameters such as blood flow (320 ml/min) and weight loss rate (0.6 +/- 0.1 kg/h), while dialysis length and dialysate conductivities were different, depending on the dialysis technique. Electrolyte kinetics and acid-base balance were similar during the three periods. The dialysis efficiency for small molecules (Kt/V of urea) was similar (between 1.4 and 1.6); however, AFB seemed to show a higher beta2-microglobulin reduction rate (47.6 +/- 4 vs. 4.3 +/- 10% for AFHD and vs. 9.9 +/- 5% for BD; p < 0.001). The nutritional and hemopoietic status maintained stable, and the hemodynamic parameters were comparable during all periods. The percent blood volume reduction at the end of the treatments was not statistically different (-14.9 +/- 9.4% in AFB, -12.1 +/- 5.1% in AFHD, and -12.2 +/- 4.4% in BD), and these results could explain the similar hemodynamic behavior during the three periods. In conclusion, AFHD appears to be a safe technique which has all positive effects of AFB and the low costs of BD. In our opinion, it could be used in patients with few clinical impairments, usually treated with hemodialysis, in whom a biocompatible treatment is indicated. PMID- 15365214 TI - Endocrine response to surgical stress and postoperative confusion in depressed patients with alcohol abuse. AB - Both alcohol drinking and depression are risk factors for postoperative confusion and are associated with alteration of pituitary-adrenal function. We investigated the incidence of postoperative confusion, plasma cortisol and ACTH response to surgical stress in depressed patients with alcohol abuse. We studied sixty depressed patients with and without alcohol abuse who underwent abdominal surgery. Postoperative confusion occurred in 4 of 30 patients (13%) in depressed patients without alcohol abuse, 10 of 30 patients (33%) in depressed patients with alcohol abuse. Plasma cortisol concentrations (27.2 +/- 7.0, 28.3 +/- 8.2, 29.2 +/- 4.1, 28.0 +/- 6.3 and 27.9 +/- 5.7 microg dl(-1)) 15, 60 min after the skin incision, 60 min after the end of surgery, the next day and the third day after surgery in depressed patients with alcohol abuse were significantly higher than that (20.1 +/- 6.4, 21.7 +/- 9.6, 22.3 +/- 8.0, 21.9 +/- 6.7 and 20.3 +/- 5.4 microg dl(-1)) in depressed patients without alcohol abuse. In depressed patients with alcohol abuse, plasma cortisol concentrations (34.9 +/- 7.1, 33.2 +/- 5.8 and 33.4 +/- 5.5 microg dl(-1)) 60 min after the end of surgery, the next day and third day after surgery in postoperatively confused depressed patients were significantly higher than those (26.4 +/- 6.3, 25.4 +/- 5.0 and 25.2 +/- 4.9 microg dl(-1)) of nonconfused depressed patients. In conclusion, the incidence of postoperative confusion was significantly higher in depressed patients with alcohol abuse than in depressed patients without alcohol abuse. Increased plasma cortisol concentrations after surgery were associated with postoperative confusion in depressed patients with alcohol abuse. PMID- 15365215 TI - Serotonergic dissection of obsessive compulsive symptoms: a challenge study with m-chlorophenylpiperazine and sumatriptan. AB - We have conducted a pharmacological challenge experiment in 10 medication-free obsessive compulsive (OC) disorder (OCD) patients. We used a placebo-controlled paradigm for m-chlorophenylpiperazine (mCPP) and sumatriptan challenges. Endocrine, physiological and behavioral variables were assessed at baseline and over a 3-hour period after the challenge. Both cortisol and prolactin were significantly elevated in OCD patients following mCPP administration. Both mCPP and sumatriptan caused significant OC symptom exacerbation with the response to sumatriptan being more robust. We conclude that the 5-HT(1Dbeta) receptor may play a role in the pathophysiology of OCD. PMID- 15365216 TI - Analysis of enhancer activity of a dinucleotide repeat polymorphism in the neurotrophin-3 gene and its association with bipolar disorder. AB - Growing evidence has implicated the possible involvement of neurotrophins in the pathogenesis of functional psychoses such as schizophrenia and bipolar disorder. Previous studies reported a significant association of a dinucleotide repeat polymorphism of the neurotrophin-3 (NTF3) gene with schizophrenia. The aims of the present study were to examine whether this polymorphism is associated with bipolar disorder and whether the polymorphic region has an enhancer/silencer effect on transcriptional activity in an allele-dependent manner. In an association analysis between the polymorphism and bipolar disorder in a Japanese sample of 88 patients and 98 controls matched for age, sex, and ethnicity, the distribution of alleles did not differ significantly between the two groups. pGL3 promoter luciferase reporter vectors containing the polymorphic region increased luciferase activity relative to empty pGL3-promoter vector in HeLa, IMR-32 (neuroblastoma) and Hs683 (glioma) cell lines; however, no significant difference was detected between alleles for either cell line. Our results suggest that the examined polymorphism has no major role in giving susceptibility to bipolar disorder. Although the polymorphic region may have an enhancer-like effect on transcriptional activity, we obtained no evidence for allele-dependent differential effects. PMID- 15365217 TI - Neuronal dysfunction of the frontal lobe in schizophrenia. AB - Localized in vivo proton magnetic resonance spectroscopy (MRS) was performed to evaluate metabolic alterations in the right and left frontal lobe before and after antipsychotic treatment of schizophrenic patients (n=24) and a group of healthy normal subjects (n=20). Proton metabolic ratios obtained from the 8 cm3 voxels in the right and left frontal lobes were compared with the clinical assessment for each subject. There was no significant difference in the metabolic ratios between the right and the left frontal lobes in either the schizophrenic group or the control group, indicating no laterality. Compared with those of the normal control group, NAA/Cr ratio of the schizophrenic patients showed significantly lower value. The NAA/Cr ratio of the schizophrenic patients was not changed after antipsychotic treatment. The present study supports the 'hypofrontality' hypothesis of schizophrenia. PMID- 15365218 TI - Familial aggregation of suicidal ideation in psychiatric patients: influence of gender. AB - To investigate the role of familial factors for suicidal ideation, 48 psychiatric patients belonging to 20 nuclear and three-generation families were personally interviewed. The effect of familiality on suicidal ideation was estimated through the use of mixed model analysis with family as random effect and gender, age and depressive score as fixed effects (covariates). The effect of family was found to be statistically significant (p=0.024) for the whole group of patients and for the subgroup of males (p=0.048), but not female patients; significance persisted for the whole group (p=0.030) and was reduced to indicative level for males (p=0.069), when psychiatric diagnosis was also considered as an additional potentially confounding factor. These results show that the occurrence of suicidal ideation has a familial component, stronger among males than females. This may be an additional explanation (besides those related to various other biological and sociocultural factors) for gender differences in suicidal behavior. PMID- 15365219 TI - Absence of NK1 receptors in human blood lymphocytes and granulocytes. AB - Substance P is a peptide that exerts its activity through the interaction with specific receptors that are distributed in different brain areas. Given the potential of NK1 receptor antagonists as antidepressants, the availability of a peripheral model of NK1 receptors would be particularly relevant for the possibility to perform studies in samples of patients. Therefore, with the present study we aimed to explore the possible existence of NK1 receptors by means of [3H]SR140333 and [125I]BHSP that behave as, respectively, antagonist and agonist, at this level, in human blood lymphocytes and granulocytes of healthy donors. The results of the present study failed to detect the presence of a high affinity and saturable binding of [3H]SR140333 and [125I]BHSP in human blood cells, whereas a specific binding for both compounds was found in rat cerebral cortex that was used as the control tissue. These findings would question the presence of NK1 receptors in human circulating cells. PMID- 15365220 TI - GABA transporters GAT-1 and GAT-3 in the human dorsolateral prefrontal cortex in schizophrenia. AB - This study aimed to investigate the binding affinity of [3H]GABA and [3H]beta alanine to GABA transporters GAT-1 and GAT-3 in the human dorsolateral prefrontal cortex (Brodmanns' area 9) in schizophrenia. Using post mortem tissue from individuals diagnosed with schizophrenia (n=6) and control subjects (n=6), the density of GAT-1 was established by displacing [3H]GABA with muscimol, and for GAT-3 [3H]beta-alanine was used. Data analysis showed a significant decrease of GAT-1 levels (45%), and a significant increase of GAT-3 density (23%) within the dorsolateral prefrontal cortex of individuals diagnosed with schizophrenia when compared to age- and sex-matched controls. The observed decrease of GAT-1 could be explained as a consequence of the GABA hypo-function or the result of volumetric shrinkage of the cerebral cortex previously reported in this disease. The observed elevation of GAT-3 levels could be due to a compensatory effect for any functional loss of GABA re-uptake by the decreased GAT-1 levels. PMID- 15365221 TI - Replication test for association of the IL-1 receptor antagonist gene, IL1RN, with attention-deficit/hyperactivity disorder. AB - Attention-deficit/hyperactivity disorder (ADHD) has a strong genetic basis, and aberrant brain dopaminergic and noradrenergic activity is implicated in its etiology. Interleukin-1 (IL-1), its antagonist, IL-1Ra, and IL-1 receptors are all present in the brain, and IL-1 has been shown to influence both dopaminergic and noradrenergic function. Recently, Segman et al. [1] tested the IL-1Ra gene, IL1RN, as a candidate for involvement in ADHD. Using the transmission/disequilibrium test (TDT) to examine 77 nuclear ADHD families for the inheritance of alleles of an intronic 86-bp VNTR polymorphism, they found significant evidence for biased transmission of the 4-repeat allele (p=0.04) and non-transmission of the 2-repeat allele (p=0.03). Here, we sought to replicate this in an independent sample of families. In contrast to the previous findings, our analysis of 178 ADHD families showed no evidence for biased transmission of these alleles (p=0.81 and p=1.00, respectively). Our lack of evidence for association of this IL1RN polymorphism with ADHD, based on a much larger sample of families, suggests that the original finding may have been a spurious (i.e. false-positive) result. These findings highlight the need for further investigations of this marker, in additional independent ADHD samples, in the future. PMID- 15365222 TI - Association study of serotonin 1B receptor (A-161T) genetic polymorphism and suicidal behaviors and response to fluoxetine in major depressive disorder. AB - Serotonin 1B receptors (5-HT1B) are autoreceptors involved in the local inhibitory control of serotonin release, and have been suggested to play a role in the pathogenesis of major depressive disorder (MDD) and the antidepressant effects of the selective serotonin reuptake inhibitors in patients. We genotyped the 5-HT1B A-161T polymorphism in 160 patients with MDD and 160 normal controls. We then tested the hypothesis that the allelic variant, A-161T, of the 5-HT1B gene confers susceptibility to MDD or is associated with suicide attempt. We also examined the association of this polymorphism with therapeutic response in 116 of the MDD patients who received fluoxetine treatment for 4 weeks. No significant difference was found in the A-161T genetic polymorphism between MDD patients and controls. The genotype distribution between patients with and without suicide attempt, or between fluoxetine treatment responders and nonresponders were also similar. Our findings suggest that 5-HT1B A-161T genetic polymorphism does not play a major role in the susceptibility to MDD, nor is it related to suicidal attempt or the therapeutic response to fluoxetine in MDD. PMID- 15365223 TI - Neuropeptide y in euthymic lithium-treated women with bipolar disorder. AB - Plasma neuropeptide Y-like immunoreactivity (NPY-LI) and platelet cyclic AMP (cAMP) activity were determined in 13 women with bipolar disorder stabilized on lithium (Li) and 12 matched healthy controls. No differences in plasma NPY-LI were found between the two groups. In euthymic Li-treated bipolar patients, there was an inverse correlation between plasma NPY-LI levels and intracellular cAMP in prostaglandin E1-stimulated platelets. A positive correlation was found between plasma NPY-LI levels and age in both the patient and the control group. Our findings support earlier findings that NPY is capable of inhibiting adenylyl cyclase and that aging is a physiological factor in regulating NPY-LI levels. PMID- 15365224 TI - Platelet 5-HT uptake in boys with conduct disorder. AB - Dysregulation of serotonergic function has been found to be associated with aggression in animals, human adults and adolescents. However, studies with children have shown conflicting results. The objective of this study was to investigate whether the kinetic characteristics (Vmax and Km) of 5-HT uptake in platelets are different in children with the diagnosis of conduct disorder according to ICD-10 and healthy age-matched controls. In addition to the standardized assessment of general psychopathology, methods assessing narrowband aggressive symptoms (Child Behavior Checklist) and emotional reactivity to an experimentally induced provocation (Taylor's competitive reaction time task) were used in both groups. We found a trend for a lower mean Vmax of platelet 5-HT uptake in 14 conduct-disordered boys compared with healthy controls (n=15). If, however, 2 patients with a low degree of aggression and emotional reactivity were excluded, the difference became significant (mean=4.27, SD=3.49 in patients and mean=8.45, SD=4.63 in controls). A significant negative correlation was found between parent-rated aggression scores and Vmax (r=-0.41, p < 0.05, n=29). These data suggest that dysfunction of 5-HT transport mechanisms might be associated with specific behavioral symptoms in conduct-disordered children. PMID- 15365225 TI - Antidepressant treatment and dehydroepiandrosterone sulfate: different effects of amitriptyline and paroxetine. AB - BACKGROUND: There is evidence for activation of the hypothalamus-pituitary adrenal system in depressed patients to be associated with increased secretion of adrenal androgens. However, there is only limited information about the effect of different classes of antidepressants on the course of adrenal androgens. METHODS: Dehydroepiandrosterone sulfate (DHEA-S) serum concentrations were measured in 80 patients being treated with amitriptyline (AMI) or paroxetine (PAROX) for a period of 35 days. RESULTS: Using analysis of variance with repeated measures, we found a significant effect of treatment upon DHEA-S serum concentrations that declined more in AMI-treated (before vs. after: 1.89+/-1.16 vs. 1.46 +/- 0.96 mg/l), compared to PAROX-treated patients (1.56+/-1.09 vs. 1.50+/-1.04 mg/l). CONCLUSIONS: Our results show changes in DHEA-S serum concentrations during antidepressant treatment to depend on medication. PMID- 15365226 TI - Propofol-induced alpha rhythm. AB - The electroencephalographic effects of two intravenous sedative/hypnotic drugs, propofol and thiopental, were studied at three stable blood concentrations in 52 normal healthy volunteers. The higher concentration resulted in unresponsiveness (lack of response to auditory/tactile stimuli) in all subjects. This report describes the strong frontal-central rhythms apparent in this state using a quantitative description of oscillatory systems underlying the rhythm. These rhythms occur when sedative drug concentrations are greater than those producing the well-described increase in broadband beta-power associated with many sedative drugs. Propofol induces rhythms in the alpha-range, while thiopental produces rhythms in the beta-range. Quasistationary for a period of about 1 h, these rhythms exceed the baseline alpha-rhythm in power. By their resonant nature, these propofol-induced rhythms are analogous to 'the classic alpha-rhythm', but quantitative characteristics of the underlying oscillatory systems are different. Baseline properties of the oscillatory system underlying the initial resting alpha-rhythm recover completely as drug concentration decays to negligible values. PMID- 15365227 TI - Fluctuations of source locations of EEG activity during transition from alertness to sleep in Alzheimer's disease and vascular dementia. AB - Vascular dementia (VaD) differs from Alzheimer's disease (AD) in larger fluctuations of cognitive impairment, hypothetically because of deteriorated vigilance control. Vigilance levels are reflected by locations of EEG sources. Transition from alertness to sleep might be particularly sensitive to degradations of vigilance control. Twelve AD and 12 VaD patients (medication free, mean age 75.6 and 77.6 years, respectively, difference = n.s.), and 12 healthy elderly subjects (mean age 70.6 years), who served as controls, were studied (each group comprised 2 males and 10 females). A twenty-one-channel EEG was recorded from full alertness to the onset of sleep stage 2. Dipole source modeling, based on Fast Fourier Transform dipole approximation, yielded 3D source localizations in 7 EEG frequency bands. For each brain axis, means of source location differences between successive 20-second periods were calculated (fluctuation magnitude). EEG band-wise MANCOVAs (3 brain axes, 3 subject groups, covariate: age) showed differences in fluctuation magnitude between groups in the 10.5- to 12-Hz alpha(2) frequency band (p=0.0066). Post hoc ANCOVAs for the axes (3 subject groups, covariate: age) were significant on the superior-inferior axis: VaD patients had higher fluctuations than AD patients and controls, without significant difference between the latter two. Thus, larger source fluctuations in VaD might reflect the patients' decreased vigilance control, accounting for their increased fluctuations of cognitive impairment. PMID- 15365228 TI - Fungal infection but not type of bacterial infection is associated with a high mortality in primary and secondary infected pancreatic necrosis. AB - INTRODUCTION: Knowledge of microbiology in the prognosis of patients with necrotizing pancreatitis is incomplete. AIM: This study compared outcomes based on primary and secondary infection after surgery for pancreatic necrosis. METHOD: From a limited prospective database of pancreatic necrosectomy, a retrospective case note review was performed (October 1996 to April 2003). RESULTS: 55 of 73 patients had infected pancreatic necrosis at the first necrosectomy. 25 of 47 patients had resistant bacteria to prophylactic antibiotics (n = 21) or did not receive prophylactic antibiotics (n = 4), but this was not associated with a higher mortality (9 of 25) compared to those with sensitive organisms (4 of 22). Patients with fungal infection (n = 6) had a higher initial median (95% CI) APACHE II score compared to those without (11 (9-13) verus 8.5 (7-10), p = 0.027). Five of six patients with fungal infection died compared to 13 of 47 who did not (p = 0.014). With the inclusion of secondary infections 21 (32%) of 66 patients had fungal infection with 10 (48%) deaths compared to 11 (24%) of 45 patients without fungal infection (p = 0.047). CONCLUSION: Whether associated with primary or secondary infected pancreatic necrosis, fungal but not bacterial infection was associated with a high mortality. PMID- 15365229 TI - Laparoscopy and laparoscopic ultrasound in the evaluation of pancreatic and periampullary tumours. AB - BACKGROUND AND AIMS: The pre-operative determination of resectability of pancreatic and peri-ampullary neoplasia assists the selection of patients for surgical or non-surgical treatment. This study investigated whether the addition of laparoscopy with laparoscopic ultrasound to dual-phase helical CT could improve the accuracy of assessment of resectability. PATIENTS AND METHODS: Prospective study of 305 patients referred to a single unit for consideration of pancreatic resection who underwent dual-phase helical CT scanning +/- laparoscopy with laparoscopic ultrasound. Data were collected on patient demographics, CT findings, assessment of operability, laparoscopic assessment (LA), surgical procedures and histology. RESULTS: LA was undertaken in 239/305 patients, 190 of whom were considered CT resectable, and 49 CT unresectable. Of the 190 CT resectable patients, LA correctly identified unresectability in 28 (15%: metastases in 15; vascular encasement in 6; anaesthesia for laparoscopy found 7 unfit for major resection) and incorrectly in 2 (vascular encasement), but did not identify unresectability in 33; LA correctly confirmed resectability in the remainder (prediction improved, chi(2) = 9.73, p < 0.01). Of the 49 CT unresectable patients, LA correctly identified resectability in 4, and incorrectly in 12, and correctly identified unresectability in the remaining 33. Sixty-six of the 305 patients did not undergo LA, of whom 23 underwent resection. CONCLUSION: When added to dual-phase helical CT, laparoscopy with laparoscopic ultrasound provides valuable information that significantly improves the selection of patients for surgical or non-surgical treatment. PMID- 15365230 TI - Percutaneous radiofrequency ablation of colorectal hepatic metastases - initial experience. An adjunct technique to systemic chemotherapy for those with inoperable colorectal hepatic metastases. AB - BACKGROUND AND AIM: Most patients with hepatic metastases from colorectal carcinoma are unsuitable for resection. Radiofrequency ablation (RFA) has been applied to such lesions at laparotomy. This study aimed to evaluate the less invasive approach of percutaneous RFA. METHOD: Patients with unresectable liver metastases identified on cross-sectional imaging were considered for percutaneous RFA either alone or in combination with systemic chemotherapy. Subjects with >6 lesions or lesions of maximum size >70 mm were excluded. Percutaneous RFA was applied under sedation and radiological guidance (CT/US). Treatment effect was determined by follow-up imaging. Actuarial survival was calculated by the Kaplan Meier analysis. RESULTS: Thirty patients (21 males), median age 74.5 years (range 44-85 years), underwent percutaneous RFA to 56 lesions during 54 treatment sessions. The median size of lesion was 30 mm (range 8-70 mm). Fifteen lesions were treated more than once because of recurrence or incomplete ablation. The median ablation time per lesion was 12 min (range 4.5-36 min). Eleven patients had pre-procedural chemotherapy and 15 patients received chemotherapy after treatment. There was minimal associated morbidity (5.6% of treatments). Median hospital stay per treatment was 1 day (range 1-7). Median actuarial survival from the date of first percutaneous RFA was 22 months (95% CI 12.9-31.1 months). Eleven patients were alive at the time of data collection. CONCLUSION: Percutaneous RFA is a safe, well-tolerated intervention for unresectable hepatic metastases which can be repeated, if required. The technique may be associated with prolonged survival in this selected group of subjects. Future studies should consider the role of percutaneous RFA either in place of or as an adjunct to palliative chemotherapy. PMID- 15365231 TI - Changes of gastrointestinal argyrophil endocrine cells in the osteoporotic SD rats induced by ovariectomy. AB - The regional distributions and frequencies of argyrophil endocrine cells in gastrointestinal (GI) tract of osteoporotic Sprague-Dawley rat induced by ovariectomy were studied using Grimelius silver stain. The experimental animals were divided into two groups, one is non-ovariectomized group (Sham) and the other is ovariectomized group (OVX). Samples were collected from each part of GI tract (fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) at 10th week after ovariectomy or sham operation. In this study, argyrophil cells were detected throughout the entire GI tract with various frequencies regardless of ovariectomy. Most of these argyrophil cells in the mucosa of GI tract were generally spherical or spindle in shape (open type cell) while cells showing round in shape (close type cell) were found occasionally in gastric and/or intestinal gland regions. The regional distributions of GI argyrophil endocrine cells in OVX were similar to those of Sham. However, significant decreases of argyrophil cells were detected in OVX compared to those of Sham except for the pylorus, jejunum and cecum. In pylorus and jejunum, argyrophil cells in OVX dramatically decreased compared to those of Sham but significances were not recorded. In addition, argyrophil cells in cecum of OVX showed similar frequency compared to that of Sham. The endocrine cells are the anatomical units responsible for the production of gut hormones that regulate gut motility and digestion including absorption, and a change in their density would reflect the change in the capacity of producing these hormones and regulating gut motility and digestion. Ovariectomy induced severe quantitative changes of GI argyrophil endocrine cell density, and the abnormality in density of GI endocrine cells may contribute to the development of gastrointestinal symptoms in osteoporosis such as impairments of calcium and some lipids, frequently encountered in patients with postmenopausal osteoporosis. PMID- 15365232 TI - Electroacupuncture ameliorates experimental colitis induced by acetic acid in rat. AB - The effect of electroacupuncture (EA) on experimental colitis was investigated in Sprague-Dawley rats. Colitis was induced by intracolonic instillation of 4% acetic acid. EA (2 Hz, 0.05 ms, 2 V for 20 min) was applied to bilateral Hoku (LI 4) and Zusanli (ST-36) on 12 hrs and 36 hrs after induction of colitis. EA treatment significantly reduced the macroscopic damage and the myeloperoxidase activity of colonic samples at 3 days post-induction of colitis. Colitic colon showed a decreased in vitro motility. However, colonic motility of EA-treated group was not significantly different from that of normal group. The anti inflammatory effect of EA was not inhibited by a glucocorticoid receptor antagonist, RU-486, but suppressed by a beta-adrenoceptor antagonist, propranonol. These results suggest that EA-treatment has a beneficial effect on colitis, and its anti-inflammatory effect is mediated by beta-adrenoceptor activation but not by endogenous glucocorticoiddependent mechanism. PMID- 15365233 TI - Molecular characterization of full-length genome of Japanese encephalitis virus (KV1899) isolated from pigs in Korea. AB - We have determined the complete nucleotide and deduced amino acid sequences of the Japanese encephalitis virus (JEV) strain KV1899, isolated from a fattening pig in Korea. In comparison with 22 fully sequenced JEV genomes currently available, we found that the 10,963-nucleotide RNA genome of KV1899 has a 13 nucleotide deletion in the 3' non-translated variable region and 53 unique nucleotide sequences including 3' non-translated region (NTR). Its single open reading frame has a total of 28 amino acid substitutions. Comparison of the KV1899 genomic sequence with those of the 21 fully sequenced JEV strains in published databases showed nucleotide homology ranging from 97.4% (Ishikawa strain) to 87.0% (CH2195 strain). Amino acid homology with KV1899 strain ranged from 96.4% (K94P05) to 91.0% (GP78). The KV1899 showed the highest nucleotide homology with Ishikawa strain and the highest amino acid homology with K94P05. We performed an extensive E gene based phylogenetic analysis on a selection of 41 JEV isolates available from the GenBank. Compared with Anyang strain, isolated from a pig in 1969, that is current live vaccine strain for swine in Korea, the homology of nucleotide sequence in envelope gene was only 87.1%. The prM gene of the isolate was closely related with those of Ishikawa and K94P05 strains, which were grouped into genotype I of JEV. PMID- 15365234 TI - Characterization and localization of the unique Marek's disease virus type 2 ORF873 gene product. AB - Studies on Marek's disease virus (MDV)-unique genes are important for understanding the biological nature of the virus. Based on complete DNA sequence analyses of the MDV genomes, the MDV genomes contain presumably at least five MDV unique genes, which are commonly conserved among the three MDV serotypes. A recombinant baculovirus that contains the MDV serotype 2 (MDV2)-unique gene, ORF873, under the polyhedrin promoter was constructed and designated rAcORF873. Polyclonal and monoclonal antibodies, which recognize the recombinant MDV2 ORF873 protein in Spodoptera frugiperda clone 9 (Sf9) cells infected with rAcORF873, were prepared by immunizing mice with a recombinant fusion protein expressed in Escherichia coli. Immunoblot analyses with the antibodies revealed a major protein band with a molecular mass of 108-kDa in both MDV2-infected chick embryo fibroblasts (CEF) and rAcORF873-infected Sf9 cells. By indirect immunofluorescence analyses using monoclonal antibody, the authentic ORF873 protein was localized in the cytoplasm of MDV2-infected CEF cells. The monoclonal and polyclonal sera, which were generated in the present study and reacted effectively to MDV2 ORF873 protein, are considered to be useful reagents for further studying the role(s) of the ORF873 protein in MDV2 infection. PMID- 15365235 TI - A comparative study of gastrointestinal parasites between ranched and free ranging Burchell's zebra (Equus burchelli antiquorum) in Isiolo district, Kenya. AB - Parasites were collected from 20 Burchell's zebra, Equus burchelli antiquorum, from Isiolo district, Kenya. 10 were ranched animals from the Lewa Downs Ranch and 10 free ranging from the areas adjacent to the ranch to the north. The animals were culled from 4th to 18th June 1995. The gastrointestinal tract was removed from the carcass and separated as stomach, small and large intestines and searched for parasites. The abdominal cavity was scrutinized for Setaria species. The parasites were identified to genus and in some cases species level. All animals were infected with at least three genera of parasites of which at least one genus was a nematode. A total of 10 genera representing eight families were recovered from the two groups. These included six nematode families, Strongylidae, Atractidae, Oxyuridae, Spiruridae, Setariidae, and Ascaridae, one cestode family, Anoplocephalidae and one family of the larvae of Gasterophilus bot flies, Gasterophilidae. The most prevalent families were Atractidae (100%) and Gasterophilidae (100%). The principle nematode genera was Crossocephalus whose total burden, plus or minus the standard deviation, was 3,471,129 +/- 1,352,922. Setaria species were recovered from the abdominal cavity of 45% of all the animals examined. The overall total worm burden was higher in the free ranging zebra 2,743,410 +/- 849,604 than the ranched zebra, 787,669 +/- 246,006. The range of individual genera varied from 0 to 269,225 in the free ranging group, which was higher than 0 to 77,890 in the ranched animals. From statistical analysis, no significant difference could be found between males and females. However, the burdens of genera Strongylus, Triodontophorus, Crossocephalus and Parascaris were significantly higher in free ranging animals. PMID- 15365236 TI - Mucosal mast cell-derived chondroitin sulphate levels in and worm expulsion from FcRgamma-knockout mice following oral challenge with Strongyloides venezuelensis. AB - Mucosal mast cell-derived chondroitin sulphates (sulphated proteoglycans) were assayed in gut washings and homogenate of FcRgamma-knockout (KO) and wild-type (WT) C57BL/6 mice challenged with Strongyloides venezuelensis in order to assess their possible role in secondary immunity against enteric nematodes. Groups of immune KO and WT mice were challenged by oral gavage with 300 infective larvae (L3). Establishment of infection was assessed by daily faecal analysis to determine the number of eggs per gram of faeces (EPG) and by adult worm recovery on days 5 and 13 post challenge. Mucosal mast cell (MMC) counts were done on days 5 and 13 post challenge while MMC-derived chondroitin sulphates in gut washings (days 1 and 5) and homogenate (day 8) were assayed by high performance liquid chromatography (HPLC). Results showed that patent infection occurred in challenged KO but not WT mice despite significantly higher mastocytosis in jejunal sections of KO than WT mice (p<0.001). Similarly but against prediction, significantly higher concentration of MMC-derived chondroitin sulphates was observed in gut homogenate of KO than WT mice (p<0.05). In contrast, significantly higher concentration of chondroitin sulphates was observed in gut washings of WT than KO mice (p<0.05). These results suggest that MMC in KO mice failed to release sufficient amount of sulphated proteoglycans into the gut lumen as did the WT mice, which may have been part of the hostile environment that prevented the establishment in and eventual expulsion of adult S. venezuelensis from the gut of WT mice following challenge. PMID- 15365237 TI - Exposure to genistein does not adversely affect the reproductive system in adult male mice adapted to a soy-based commercial diet. AB - Genistein, a soybean-originated isoflavone, is widely consumed by humans for putative beneficial health effects but its estrogenic activity may affect adversely the development of male reproductive system. Five-week-old ICR mice were purchased and fed with a soybean-based Purina Chow diet until 6 months of age. The animals were exposed by gavage to genistein (2.5 mg/kg/day) or 17beta estradiol (7.5 microg/kg/day) for five weeks. Corn oil was used for the negative control. The animals were fed the casein-based AIN-76A diet throughout the experimental periods. There were no significant differences in body and organ weights of mice among experimental groups. No significant differences in sperm counts and sperm motile characteristics were found between the control and the genistein groups. Treatment of 17beta-estradiol caused a significant decrease in epididymal sperm counts compared to the control (p<0.05). The level of phospholipid hydroxide glutathione peroxidase in the epididymis of mice exposed to genistein was significantly higher than that of the control mice (p<0.05). 17beta-estradiol treatment caused a reduction of germ cells in the testis and hyperplasia of mucosal fold region in the prostate of mice. Genistein treatment did not cause any lesion in the testis, epididymis, and prostate. These results suggest that dietary uptake of genistein at adult stage of life may not affect male reproductive system and functions. PMID- 15365238 TI - An ultrastructural study on cytotoxic effects of mono(2-ethylhexyl) phthalate (MEHP) on testes in Shiba goat in vitro. AB - In this study, the effects of mono(2-ethylhexyl) phthalate (MEHP), one of metabolites of di(2-ethylhexyl) phthalate, on immature Shiba goat testes in vitro were examined. The testes of 2-month-old Shiba goats were cut into smaller pieces, and seeded in medium. At 1, 3, 6 and 9 hr after administration of MEHP at various concentrations (0, 100 nmol ml(-1), 1 nmol ml(-1), and 1 x 10(-3) nmol ml(-1), respectively), the specimens were obtained for light and transmission electron microscopic observations. As a result, at 1 hr after exposure to MEHP, the vacuolization and nuclear membrane rupture appeared in Sertoli cells. Such alterations tended to gradually increase in number in time and dose-dependent manners. Moreover, by MEHP treatment, apoptotic spermatogenic cells (characterized with chromatin condensation, cytoplasm shrinkage without membrane rupture, still functioning cell organelles, and packed cell contents in membrane bounded bodies), apoptotic Sertoli cells (characterized with nuclear membrane lysis, nuclear condensation), necrotic spermatogenic cells (characterized with swollen and ruptured mitochondria, plasma membrane lysis, spilt cell contents, and chromatin clumps), and necrotic Sertoli cells (characterized with marginated chromatins along the nuclear membrane, ruptured vesicles within the MNB, some swollen and ruptured cell organelles, e.g. mitochondria) could be identified. Conclusively, ultrastructurally the treatment with MEHP at low concentration tends to lead spermatogenic and Sertoli cells to apoptosis, whereas that at high concentration tends to lead spermatogenic and Sertoli cells to necrosis. Thus, the testicular tissue culture is advantageous for screening testicular toxicity of chemicals. PMID- 15365239 TI - Repeated dose toxicity of alfa-cypermethrin in rats. AB - The present study was performed to investigate the subacute effect of alpha cypermethrin (alpha-CP) in rats. Alfacypermethrin a synthetic pyrethroid insecticide, dissolved in dimethyl sulfoxide (DMSO) and oral LD50 was investigated after administering orally different doses in rats and was determined as 145 mg/kg. Other groups of rats were given repeated daily oral dose (1/10 LD50) of alpha-CP for 30 days. The animals were sacrificed on 31st day. Activities of various enzymes, cytochrome P450 and b5 contents in liver, hepatic antioxidant status, tissue residue concentration, haemogram and pathological changes were studied. It increased the serum aminotransaminases (AST, ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities and blood glucose level significantly. alpha-CP decreased RBC count, PCV and Hb level significantly. It significantly decreased cytochrome P450 in liver. Residues were present in different tissues. It increased malondialdehyde (MDA) level, while decreased the activities of catalase (CAT), superoxide dismutase (SOD) and glycogen level in liver significantly. Mild to moderate histological alterations were observed in lungs, liver, stomach, kidneys, testes and cerebellum. So repeated daily oral doses of alpha-CP at 1/10LD50 altered the biochemical parameters, decreased cytochrome P450 content, antioxidant status, which correlated with histopathological changes of tissues. PMID- 15365240 TI - Thickness of cumulus cell layer is a significant factor in meiotic competence of buffalo oocytes. AB - This study evaluated the meiotic competence of buffalo oocytes with different layers of cumulus cells. A total of 588 oocytes were collected from 775 ovaries averaging 0.78 oocytes per ovary. Oocytes with homogenous cytoplasm (n = 441) were selected for in vitro maturation (IVM) and divided into four groups based on their cumulus morphology: a) oocytes with > or = = 3 layers of cumulus cells, b) 1-2 layers of cumulus cells and oocytes with partial remnants or no cumulus cells to be cocultured c) with or d) without cumulus cells. Oocytes in all four groups were matured in 100 microL drop of TCM-199 supplemented with 10 microg/mL follicle stimulating hormone (FSH), 10 microg/mL luteinizing hormone (LH), 1.5 microg/mL estradiol, 75 microg/mL streptomycin, 100 IU/mL penicillin, 10 mM Hepes and 10% FBS at 39 degrees C and 5% CO2 for 24 hours. After IVM, cumulus cells were removed from oocytes using 3 mg/mL hyaluronidase, fixed in 3% glutaraldehyde, stained with DAPI and evaluated for meiotic competence. The oocytes with > or = 3 layers of cumulus cells showed higher maturation rates (p<0.05: 64.5%) than oocytes with partial or no cumulus cells (8.6%) and oocytes co-cultured with cumulus cells (34.5%) but did not differ from oocytes having 1-2 layers of cumulus cells (51.4%). The degeneration rates were higher (p<0.05) for oocytes with partial or no cumulus cells (51%) than rest of the groups (range: 13.8% to 17.4%). These results suggest that buffalo oocytes with intact layers of cumulus cells show better IVM rates than oocytes without cumulus cells and the co culture of poor quality oocytes with cumulus cells improves their meiotic competence. PMID- 15365241 TI - Effect of beta-mercaptoethanol or epidermal growth factor supplementation on in vitro maturation of canine oocytes collected from dogs with different stages of the estrus cycle. AB - Supplementation of beta-mercaptoethanol (beta-ME) in in vitro maturation (IVM) medium was shown to improve embryo development and quality in several species. Epidermal growth factor (EGF) was also shown to improve IVM of human oocyte and embryo development after in vitro fertilization (IVF). The effect of these two compounds were suggested to be mediated through the synthesis of glutathione (GSH) which is known to play an important role in protecting the cell or embryos from oxidative damage. Thus, it is suggested that supplementation of canine IVM medium with beta-ME or EGF may be of benefit due to its positive role in IVM of various mammalian oocytes and embryo development, including cattle, pigs, rodents and humans. This study investigates the effect of ovarian estrus stage on canine oocyte quality and supplementation of medium with beta-ME or EGF on IVM of canine oocytes. As results, a significantly higher percentage of oocytes progressed to metaphase II (MII) stage in 50 or 100 microM of beta-ME supplemented oocytes collected from the follicular stage. The maturation rate to metaphase I (MI) stage was also significantly higher in oocytes collected from follicular stage and cultured with 25 or 100 microM compared to other experimental groups. After IVM culture, oocytes recovered from dogs with the follicular stage and matured in TCM-199 supplemented with 20 ng/ml EGF yielded better oocyte maturation to MII phase compared to other groups. Taken together, supplementation of beta-ME (50 or 100 microM) or EGF (20 ng/ml) improved IVM of canine oocytes to MII stage. PMID- 15365242 TI - Echocardiography and electrocardiography as means to evaluate potential performance in horses. AB - Prediction of potential performance is one of the goals of exercise physiology investigations. When Selecting a horse for competition, one of the main objectives is to choose the one that predictably will reveal a competitive aptitude above the average. The horses used in this study underwent a two dimensional echocardiography study and a conventional 3 leads electrocardiogram. The results show that heart score is not an appropriate index to evaluate the heart size in the horse. On the other hand, there are currently more suitable and accurate procedures such as echocardiography that allow performing a clear anatomical evaluation and accurate measurement in order to calculate LVMM and to predict performance. PMID- 15365243 TI - Persistent occurrence of a single Streptococcus equi subsp. zooepidemicus clone in the pig and monkey population in Indonesia. AB - In the present study 41 mucoid growing Streptococcus equi subsp. zooepidemicus strains (37 strains isolated from healthy two from diseased pigs, two strains isolated from healthy monkeys) appeared to be phenotypically and genotypically identical to mucoid growing S. equi subsp. zooepidemicus strains isolated from a previously described outbreak among the pig and monkey population on the island of Bali, Indonesia. These findings indicate that the mucoid growing S. equi subsp. zooepidemicus clone was still present in the pig and monkey population in Indonesia. PMID- 15365244 TI - Use of the n-butyl cyanoacrylate adhesive and the polyglactine thread suture for corneal rhaphy in rabbit (Oryctolagus cunicullus). AB - The aim of this study was to evaluate the cicatricial repair of perforating cornea in rabbits, by using the N-butyl cyanoacrylate adhesive compared to the 910-polyglactine thread suture through macroscopic and histological assays. Corneas from 18 adult rabbits were perforated and subsequently occluded with N butyl cyanoacrylate synthetic adhesive (right cornea) or by separated single points using the 910-polyglactine thread (left cornea). The rabbits were divided into groups containing three animals per group. Examination after 7, 15, and 30 days post-operative showed that both the synthetic adhesive and the suture were efficient in the occlusion of the surgical wounds, thus stabilizing the intra ocular content. The N-butyl cyanoacrylate adhesive was shown to be superior to the 910-polyglactine suture thread with regards to the evolution and the organization of the healing process. PMID- 15365245 TI - Glucocorticoid-induced laminitis with hepatopathy in a Thoroughbred filly. AB - A 3-year-old Thoroughbred filly was referred to the Equine Hospital, Korea Racing Association for evaluation of hematuria, inappetite, weight loss and depression. From 25 days prior to admission, the horse was treated for right carpal lameness with 20 mg intramuscular administration of triamcinolone acetonide per day for consecutive 10 days by a local veterinarian. Clinical and laboratory findings included vaginal hyperemia, flare in bladder wall, neutrophilia, lymphopenia, polyuria, polydipsia and laminitis in the end. High activities of aspartate transaminase and gamma glutamyltransferase and high concentration of total bilirubin indicated hepatopathy. Further hematology, serum biochemistry and urinalysis did not reveal any abnormalities. Medical history, physical and clinicopathologic findings suggest that the laminitis and hepatopathy in this horse were most likely induced by repeated administration of exogenous corticosteroid. However, guarded prognosis of treating laminitis undermined the benefit of improvement of hematuria following electroacupuncture stimulation. The combined stimulation of kidney related acupoints (Shen Peng, Shen Shu), lumber related acupoints (Yao Qian, Yao Zhong) and associate acupoints (Guan Yuan Shu, Bai Hui) at 5Hz, 1-2V, for 40 minutes was of value in the treatment of hematuria. This case shows that horses under steroids may exhibit laminitis and steroid hepatopathy. Early recognition and good management of laminitis are important in the limitation of complications. PMID- 15365246 TI - Surgical removal of stones in the stomach of a tiger shovelnose catfish. AB - A seven months old, tiger shovelnose catfish was referred to the veterinary medical teaching hospital of the college of veterinary medicine, Konkuk university because of suspecting a foreign body in the stomach. By physical and radiologic examinations, the catfish revealed abdominal enlargement, vomiting, inactivity and radiopaque foreign bodies in the stomach. Under general anesthesia with dilute isoflurane solution, celiotomy was performed. Stomach was atonic, and three foreign bodies were palpated in the stomach. In this process, three stones came out through mouth. Abdominal muscles were closed with a simple continuous suture pattern, and skin was closed with a simple mattress suture pattern. Sutured site was covered with glue and mucous taken from allogenic skin. On the second day after surgery, the patient recovered to normal condition, showing increased appetite and activity.